PMID- 14733929 TI - IFN-gamma-induced SOCS-1 regulates STAT6-dependent eotaxin production triggered by IL-4 and TNF-alpha. AB - The production of eotaxin, which is a critical mediator for airway inflammation, is inhibited by IFN-gamma. Here, we investigated the precise mechanisms underlying IFN-gamma-dependent inhibition of eotaxin production using mouse embryonic fibroblasts (MEF). MEF produced high levels of eotaxin in STAT6 dependent manner when they were cultured with both IL-4 and TNF-alpha. However, the eotaxin production by MEF was strongly inhibited by addition of IFN-gamma. Western-blotting analysis demonstrated that IFN-gamma downmodulated STAT6 phosphorylation induced by IL-4 and TNF-alpha. Moreover, IFN-gamma did not exhibit its inhibitory effect on both STAT6-phosphorylation and eotaxin production in MEF obtained from deficient mice in STAT1, a key molecule of IFN gamma signaling. We also demonstrated that SOCS-1, a potent inhibitory molecule of IL-4 signaling, was induced by IFN-gamma in STAT1-dependent manner. This indicated that SOCS-1 might be involved in IFN-gamma-mediated STAT1-dependent inhibition of eotaxin production. In SOCS-1(-/-) MEF, IFN-gamma inhibited neither STAT6 phosphorylation nor eotaxin production induced by IL-4 and TNF-alpha. Conversely, retroviral transduction of SOCS-1 into MEF inhibited STAT6 phosphorylation and eotaxin production induced by IL-4 and TNF-alpha, in the absence of IFN-gamma. Thus, we demonstrated that IFN-gamma-induced inhibition of STAT6 phosphorylation and eotaxin production were mediated by SOCS-1 induced in STAT1-dependent manner. PMID- 14733930 TI - N(6)-Methyldeoxyadenosine, a nucleoside commonly found in prokaryotes, induces C2C12 myogenic differentiation. AB - N(6)-methyl-2(')-deoxyadenosine (MedAdo) is a nucleoside naturally found in prokaryotic DNA. Interestingly, the N(6)-methylation of adenine in DNA seems to have been counter-selected during the course of evolution since MedAdo has not been detected in mammalian DNA until now. We show here that treatment with MedAdo induces myogenesis in C2C12 myoblasts. The presence of MedAdo in C2C12 DNA was investigated using a method based on HPLC coupled to electrospray ionization tandem mass spectrometry which is several thousand fold more sensitive than assays used previously. By this procedure, MedAdo is detected in the DNA from MedAdo-treated cells but remains undetectable in the DNA from control cells. Furthermore, MedAdo regulates the expression of p21, myogenin, mTOR, and MHC. Interestingly, in the pluripotent C2C12 cell line, MedAdo drives the differentiation towards myogenesis only. Thus, the biological effect of MedAdo is suppressed in the presence of BMP-2 which transdifferentiates C2C12 from myogenic into osteogenic lineage cells. Taken together these results point to MedAdo as a novel inducer of myogenesis and further extends the differentiation potentialities of this methylated nucleoside. Furthermore, these data raise the intriguing possibility that the biological effects of MedAdo on cell differentiation may have led to its counter-selection in eukaryotes. PMID- 14733931 TI - Enhanced neurite outgrowth by human neurons grown on solid three-dimensional scaffolds. AB - Growing and differentiating human stem cells in vitro can provide access to study the molecular mechanisms that control cellular development in a manner pertinent to human embryogenesis. To fully understand such processes, however, it is important to recreate culture conditions that most closely relate to those in living tissues. As step in this direction, we have developed a robust three dimensional cell culture system using inert highly porous solid matrices manufactured from polystyrene that can be routinely used to study the differentiation of human pluripotent stem cell-derived neurons in vitro. Neurite outgrowth was significantly enhanced when neurons were grown in a three dimensional environment compared to traditional flat surfaces and resulted in the formation of extensive neural networks. These data suggest that the topography within the culture environment can significantly alter cell development and will therefore be an important feature when investigating the potential of human stem cells. PMID- 14733932 TI - Cloning of a gene encoding 4-amino-3-hydroxybenzoate 2,3-dioxygenase from Bordetella sp. 10d. AB - Bordetella sp. 10d produces a novel dioxygenase catalyzing the meta-cleavage of 4 amino-3-hydroxybenzoic acid, 4-amino-3-hydroxybenzoate 2,3-dioxygenase (4A3HBA23D). A gene encoding 4A3HBA23D was cloned and named ahdA. The deduced amino acid sequence of ahdA showed 29.2-24.2% identities to those of prokaryotic and eukaryotic 3-hydoxybenzoate 3,4-dioxygenases in reported meta-cleavage dioxygenases. However, no identities were observed in the amino-terminal sequences of the first 29 amino acid residues. An ORF was found downstream of ahdA. The deduced amino acid sequence of the ORF showed identities to those of LysR family regulators involved in protocatechuate metabolism and contained motifs conserved in the regulators. On the basis of these results, the ORF was named ahdR encoding a putative LysR family regulator. The transcription start point of ahdA was localized 414-bp upstream of the start codon of ahdA. Two DNA binding motifs of LysR family regulators were found upstream of the transcription start point. These observations suggest that a LysR family regulator encoded by ahdR regulates the expression of ahdA. PMID- 14733933 TI - Collaborated regulation of female-specific murine Cyp3a41 gene expression by growth and glucocorticoid hormones. AB - CYP3A41 is a female-specific major CYP3A in mouse livers. Adrenalectomy decreased expression of CYP3A41 as well as CYP3A11, another major CYP3A, and dexamethasone (DEX) restored the decreased expression. Hypophysectomy completely abolished CYP3A41 expression and growth hormone (GH) replacement only slightly restored the expression. Treatment with DEX alone did not induce expression of either CYP3A41 or CYP3A11 in hypophysectomized mice. However, combined treatment with GH and DEX strongly induced expression of CYP3A41 but not CYP3A11. In primary cultured mouse hepatocytes, DEX induced expression of both CYP3A41 and CYP3A11, and DEX inducible expression of CYP3A41 was suppressed by RU486, a potent antiglucocorticoid. In contrast, RU486 by itself enhanced basal expression of CYP3A11 mRNA, while it showed no inhibitory effect on DEX-inducible expression. These observations indicate that glucocorticoids may participate in the GH dependent control of the Cyp3a41 gene expression, probably mediated via the glucocorticoid receptor, which may be different from that of the Cyp3a11 gene expression. PMID- 14733934 TI - Dipyridamole is neuroprotective for cultured rat embryonic cortical neurons. AB - The effects of a clinically useful cardiovascular agent, dipyridamole, were examined in a rodent tissue culture model of neuroprotection. Dipyridamole effectively protected rat embryonic day 18 (E18) cortical neurons from either 48 h trophic deprivation or 48 h exposure to the glutathione synthesis inhibitor, L buthionine (R,S) sulfoximine. The neuron sparing actions of dipyridamole were time- and concentration-dependent and mimicked the actions of exogenously applied glutathione. These results demonstrate that dipyridamole protects primary neuronal cultures against either trophic or chemically mediated insults, and suggest that dipyridamole has a potent antioxidant ability that compensates for glutathione depletion in neuronal cultures. PMID- 14733935 TI - Degradation of the Id2 developmental regulator: targeting via N-terminal ubiquitination. AB - Degradation of cellular proteins via the ubiquitin-proteasome system (UPS) involves: (i) generation of a substrate-anchored polyubiquitin degradation signal and (ii) destruction of the tagged protein by the 26S proteasome with release of free and reusable ubiquitin. For most substrates, it is believed that the first ubiquitin moiety is conjugated to a epsilon-NH(2) group of an internal Lys residue. Recent findings indicate that for several proteins, the first ubiquitin moiety is fused, in a linear manner, to the free alpha-NH(2) group of the protein. Here, we demonstrate that the inhibitor of differentiation (or inhibitor of DNA binding) 2, Id2, that downregulates gene expression in undifferentiated and self-renewing cells, is degraded by the UPS following ubiquitination at its N terminal residue. Lysine-less (LL) Id2 is degraded efficiently by the proteasome following ubiquitination. Fusion of a Myc tag to the N-terminal but not to the C terminal residue of Id2 stabilizes the protein. Furthermore, deletion of the first 15 N-terminal residues of Id2 stabilizes the protein, suggesting that this domain serves as a recognition element, possibly for the ubiquitin ligase, E3. The mechanisms and structural motives that govern Id2 stability may have important implications to the regulation of the protein during normal differentiation and malignant transformation. PMID- 14733936 TI - Sphingomyelinase of Helicobacter pylori-induced cytotoxicity in AGS gastric epithelial cells via activation of JNK kinase. AB - The aim of this study was to determine whether the Helicobacter pylori-derived sphigomyelinase (SMase) affects the sphingomyelin pathway and growth in AGS epithelial cells. We showed that the exogenous SMase increased the intracellular level of ceramide in AGS cells and led to rapid stimulation of extracellular signal-regulated kinase (ERK) and c-Jun kinase (JNK) activities. Incubation of AGS cells with H. pylori-derived SMase also resulted in suppression of cell growth and a concomitant induction of apoptosis. Data showed that PD98059 (up to 50 microM), an ERK inhibitor, did not affect the cell viability, whereas the cytotoxicity of exogenous SMase was completely blocked by SP600125, a JNK inhibitor at a concentration of 210 nM. We conclude that the activation of the mitogen-activated protein (MAP) kinases in AGS cells by exogenous H. pylori SMase is a major pathway to mediate the cytotoxicity. PMID- 14733937 TI - A distinctive ribonuclease from fresh fruiting bodies of the medicinal mushroom Ganoderma lucidum. AB - A ribonuclease with an N-terminal sequence distinct from other mushroom ribonucleases was isolated from fresh fruiting bodies of the medicinal mushroom Ganoderma lucidum. The ribonuclease was adsorbed on DEAE-cellulose and Q Sepharose, and unadsorbed on CM-Sepharose. It possessed a molecular mass of 42 kDa as judged by gel filtration by fast protein liquid chromatography on Superdex 75 and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Its molecular mass was similar to that of straw mushroom ribonuclease but much higher compared with those of other mushroom ribonucleases. The ribonuclease was unique among mushroom ribonucleases in that it exhibited the highest potency toward poly(U), followed by poly(A). Its activity toward poly(G) and poly(C) was about one-half of that toward poly(A) and one-quarter of that toward poly(U). A pH of 4.0 and a temperature of 60 degrees C were required for optimal activity of the enzyme. The optimum pH was low compared with those reported for other mushroom ribonucleases. PMID- 14733938 TI - Protein-protein interactions among human 20S proteasome subunits and proteassemblin. AB - Immunoproteasomes and standard proteasomes assemble by alternative pathways that bias against the formation of certain "mixed" proteasomes. Differences between beta subunit propeptides contribute to assembly specificity and an assembly chaperone, proteassemblin, may be involved via differential propeptide interactions. We investigated possible mechanisms of biased proteasome assembly and the role of proteassemblin by identifying protein-protein interactions among human 20S proteasome subunits and proteassemblin using a yeast two-hybrid interaction assay. Forty-one interactions were detected, including five involving proteassemblin and contiguous beta subunits, which suggests that proteassemblin binds to preproteasomes via a beta subunit surface. Interaction between proteassemblin and beta5, but not beta5i, suggests that proteassemblin may be involved in the propeptide-dependent differential incorporation of these subunits. Interactions between proteassemblin and beta1, beta1i, and beta7 suggest that proteassemblin may regulate preproteasome dimerization via interactions with the C-termini of these subunits, which in the mature 20S structure extend to contact opposing beta subunit rings. PMID- 14733939 TI - In vitro sliding of actin filaments labelled with single quantum dots. AB - We recently refined the in vitro motility assay for studies of actomyosin function to achieve rectified myosin induced sliding of actin filaments. This paves the way, both for detailed functional studies of actomyosin and for nanotechnological applications. In the latter applications it would be desirable to use actin filaments for transportation of cargoes (e.g., enzymes) between different predetermined locations on a chip. We here describe how single quantum dot labelling of isolated actin filaments simultaneously provides handles for cargo attachment and bright and photostable fluorescence labels facilitating cargo detection and filament tracking. Labelling was achieved with preserved actomyosin function using streptavidin-coated CdSe quantum dots (Qdots). These nanocrystals have several unique physical properties and the present work describes their first use for functional studies of isolated proteins outside the cell. The results, in addition to the nanotechnology developments, open for new types of in vitro assays of isolated biomolecules. PMID- 14733940 TI - Implication of gamma-secretase in neuregulin-induced maturation of oligodendrocytes. AB - Increasing evidences suggest that, after neuregulin (NRG) stimulation, ErbB4 undergoes a series of proteolysis, including gamma-secretase cleavage. The released ErbB4 intracellular domain (EICD) is translocated into nucleus and has a transcriptional function. Although NRG-ErbB4 signaling mediates maturation of oligodendrocytes (OLs), the role of EICD and gamma-secretase in this process remains elusive. Here, we showed that NRG-ErbB4 interaction accumulated EICD in the nucleus and promoted the expression of myelin basic protein expression in OLs. Conversely, inhibitor of ErbB4 or gamma-secretase blocked the capacity of NRG. Nuclear accumulation of EICD did not influence maturation of neurons and astrocytes and early development of OLs. We also found that EICD translocation accorded a temporal pattern, consistent with the developmental gradient of hippocampus. Our data suggest that gamma-secretase activation and EICD nuclear translocation are required for OL maturation induced by NRG, and ErbB4 acts as a functional receptor depending on a new signaling cascade. PMID- 14733941 TI - Probing protein-surfactant interaction by steady state and time-resolved fluorescence spectroscopy. AB - The microenvironment of the probe coumarin 153 (C-153) in 1% bovine serum albumin (BSA) is more hydrophobic in nature compared to that in pure micelles or protein surfactant complexes. In the native state of protein, we have not observed any solvation using C-153 as a probe but we have observed a slow dynamics on protein surface using 8-anilino-1-naphthalenesulfonic acid (ANS) as a probe. This may be due to the location of the probe (C-153) in the hydrophobic, solvent-inaccessible pocket of the BSA. Solvation dynamics in the BSA-surfactant (SDS) complexes in the solution phase is markedly different from that in pure micelles. This is may be due to the formation of 'necklace and bead' structure in the complexes. The rotational motion is also severely hindered in the surface of the protein. PMID- 14733942 TI - Cytotoxicity of RNases is increased by cationization and counteracted by K(Ca) channels. AB - K(Ca) channels are involved in control of cell proliferation and differentiation. Here we have revealed their role in overcoming the RNase-induced cytotoxicity. Toxic effects of Streptomyces aureofaciens RNases Sa, Sa2, Sa3, and of RNase Sa charge reversal mutants on the human embryonic kidney cell lines differing only by the presence of K(Ca) channels were characterized. In contrast to other RNases, a basic variant of RNase Sa and RNase Sa3 exhibit significant cytotoxic activity of the same order of magnitude as onconase. Our data indicate the absence of a correlation between catalytic activity and stability of RNases and cytotoxicity. On the other hand, cationization enhances toxic effect of an RNase indicating the major role of a positive charge. Essentially lower sensitivity to cytotoxic microbial RNases of cells expressing K(Ca) channels was found. These results suggest that cells without the K(Ca) channel activity cannot counteract toxic effect of RNases. PMID- 14733943 TI - Modulation of the redox state of tubulin by the glutathione/glutaredoxin reductase system. AB - Alterations in the redox status of proteins have been implicated in the pathology of several neurodegenerative diseases. We report that peroxynitrite-induced disulfides in porcine brain tubulin are repaired by the glutaredoxin reductase system composed of glutathione reductase, human or Escherichia coli glutaredoxin, reduced glutathione, and NADPH. Reduction of disulfide bonds between the alpha- and beta-tubulin subunits by the glutathione reductase system was assessed by Western blot. Tubulin cysteine oxidation and reduction was quantitated by monitoring the incorporation of 5-iodoacetamido-fluorescein, a thiol-specific labeling reagent. Tubulin disulfide bond reduction by the glutaredoxin reductase system restored tubulin polymerization activity that was lost following peroxynitrite addition. In support of redox modulations of tubulin by glutathione, thiol-disulfide exchange between tubulin and oxidized glutathione was detected and quantitated by HPLC. In addition, glutathionylation of tubulin was detected by dot blot using an anti-GSH antibody. PMID- 14733944 TI - Hypoxia-induced decrease of UCP3 gene expression in rat heart parallels metabolic gene switching but fails to affect mitochondrial respiratory coupling. AB - Mitochondrial uncoupling proteins 2 and 3 (UCP2 and UCP3) are postulated to contribute to antioxidant defense, nutrient partitioning, and energy efficiency in the heart. To distinguish isotype function in response to metabolic stress we measured cardiac mitochondrial function and cardiac UCP gene expression following chronic hypobaric hypoxia. Isolated mitochondrial O(2) consumption and ATP synthesis rate were reduced but respiratory coupling was unchanged compared to normoxic groups. Concurrently, left ventricular UCP3 mRNA levels were significantly decreased with hypoxia (p<0.05) while UCP2 levels remained unchanged versus controls. Diminished UCP3 expression was associated with coordinate regulation of counter-regulatory metabolic genes. From these data, we propose a role for UCP3 in the regulation of fatty acid oxidation in the heart as opposed to uncoupling of mitochondria. Moreover, the divergent hypoxia-induced regulation of UCP2 and UCP3 supports distinct mitochondrial regulatory functions of these inner mitochondrial membrane proteins in the heart in response to metabolic stress. PMID- 14733945 TI - Annexin-I expression modulates drug resistance in tumor cells. AB - The use of anti-cancer chemotherapy often leads to the rise of multidrug resistant (MDR) tumors. We have previously reported the overexpression of a 40kDa protein (P-40) in several MDR tumor cell lines. In this report we describe the cloning of a 1.4kb cDNA with an open reading frame of 344 amino acids that encodes the P-40 protein. Analysis of the P-40 amino acid sequence showed it is identical to the human annexin I (Anx-I) protein. The identity of the isolated P 40 cDNA as Anx-I was confirmed by the specific binding of IPM96 mAb to a 40kDa protein following the in vitro expression of P-40 full-length cDNA. Northern blot analysis of total RNA from drug-sensitive and -resistant cells revealed an increase in P-40 (or Anx-I) mRNA in drug-resistant cells relative to drug sensitive cells. Transfection of Anx-I cDNA into drug-sensitive MCF-7 cells was carried out without further drug selection and showed 2- to 5-fold increase in resistance of transfected cells to adriamycin, melphalan, and etoposide. Conversely, transfection of reverse Anx-I cDNA into SKOV-3 cells decreased the expression of Anx-I without affecting the expression of other members of the annexin family and showed a 3- to 8-fold increase in sensitivity to these drugs. Of interest was the correlation between the presence of Anx-I and MDR in MDA-MB 231 cells when compared to MCF-7 cells. MDA-MB-231 cells show 3- to 20-fold increase in resistance to adriamycin, melphalan, and etoposide in the absence of detectable levels of P-glycoprotein (P-gp1), the multidrug resistance protein (MRP1) or the breast cancer resistance protein (BCRP). Taken together, these results provide the first direct evidence for the role of Anx-I in MDR of tumor cells. PMID- 14733946 TI - Cdc42-dependent nuclear translocation of non-receptor tyrosine kinase, ACK. AB - Ras signals for the transformation of mammalian cells are apparently transduced through Rho GTPases. The Rho GTPase family member Cdc42 generates independent signals that regulate the rearrangement of the actin cytoskeleton and the transcription of genes. However, the molecular mechanism of signal transduction from Cdc42 to the nucleus remains to be understood. The non-receptor tyrosine kinases ACK-1 and ACK-2 have been found to bind specifically to Cdc42. In this paper we studied whether ACKs transduce Cdc42 signals to the nucleus directly, or through other cytoplasmic proteins. Using immunocytochemistry and Western blot analysis, we found a nuclear localization of ACKs in semi-confluent glioblastoma (U251) cells, as opposed to a cytosolic localization in confluent cells. In agreement with the nuclear localization, a putative nuclear export signal was identified in ACK-1 and ACK-2. Furthermore, the interaction of Cdc42 with ACKs was shown to be essential for the nuclear localization of ACKs. Overexpression of ACK42 (a Cdc42 binding domain of ACK) inhibited cell growth and movement, indicating that Cdc42 signals are transduced to the nucleus through ACKs. This is the first report providing evidence of a novel role for ACKs in transducing Cdc42 signals directly to the nucleus. PMID- 14733947 TI - Pioglitazone treatment activates AMP-activated protein kinase in rat liver and adipose tissue in vivo. AB - Thiazolidinediones have been shown to activate AMP-activated protein kinase activity in cultured cells. Whether they have a similar effect in vivo and if so whether it is physiologically relevant is not known. To assess these questions, we examined the effects of pioglitazone, administered orally to intact rats, on AMPK phosphorylation (AMPK-P) (a measure of its activation) and acetyl CoA carboxylase (ACC) activity and malonyl CoA concentration in rat liver and adipose tissue. In the first study, measurements were made in the Dahl-salt-sensitive rat (Dahl-S), a strain of Sprague-Dawley rat with endogenous hypertriglyceridemia and high levels of malonyl CoA that are restored to control values by pioglitazone. Treatment with pioglitazone (20mg/kg bw/day for 3 weeks) did not significantly increase either P-AMPK or P-ACC (which varies inversely with ACC activity) in control rats. However, in the Dahl-S rats values for AMPK-P and ACC-P were 50% lower than in control rats and were doubled by pioglitazone treatment. In a second study, the effects of two weeks treatment with pioglitazone (3mg/kg bw/day administered orally) were evaluated in Wistar rats. Under basal conditions (no manipulation of the animals), pioglitazone increased AMPK phosphorylation by twofold and decreased ACC activity and the concentration of malonyl CoA by 50% in liver. Following a euglycemic-hyperinsulinemic clamp (6h), 50% decreases in AMPK and ACC phosphorylation (indicating an increase in its activity) and comparable increases in malonyl CoA concentration were observed in liver and adipose tissue. In both tissues, pre-treatment with pioglitazone prevented these changes. Where studied (in Wistar rats under basal conditions) treatment with pioglitazone decreased the concentration of ATP by 1/3 and increased the concentration of ADP and AMP in liver. The results indicate that treatment with pioglitazone can increase AMPK activity in rat liver and adipose tissue in a variety of circumstances. They also suggest that this activation of AMPK may be mediated by a change in cellular energy state. Whether these effects of pioglitazone contribute to its insulin-sensitizing and other actions in vivo remains to be determined. PMID- 14733948 TI - Sorting pathway and molecular targeting signals for the Arabidopsis peroxin 3. AB - Peroxin 3 (Pex3p) has been identified and characterized as a peroxisomal membrane protein in yeasts and mammals. We identified two putative homologs in Arabidopsis (AtPex3p, forms 1 and 2), both with an identical cluster of positively charged amino acid residues (RKHRRK) immediately preceding one of the two predicted transmembrane domains (TMD1). In transiently transformed Arabidopsis and tobacco BY-2 suspension-cultured cells, epitope-tagged AtPex3p (form 2) sorted post translationally from the cytosol directly to peroxisomes, the first sorting pathway described for any peroxin in plants. TMD1 and RKHRRK were necessary for targeting form 2 to peroxisomes and sufficient for directing chloramphenicol acetyltransferase to peroxisomes in both cell types. The N and C termini of AtPex3p (form 2) extend into the peroxisomal matrix, different from mammal and yeast Pex3 proteins. Thus, two authentic peroxisomal membrane-bound Pex3p homologs possessing a membrane peroxisomal targeting signal, the first one defined for a plant peroxin and for any Pex3p homolog, exist in plant cells. PMID- 14733949 TI - 8Br-cGMP mediates relaxation of tracheal smooth muscle through PKA. AB - In this study, guinea pig tracheal smooth muscle pre-contracted with histamine was relaxed by the addition of 100microM 8Br-cGMP, a non-hydrolyzable and cell permeable analog for cGMP. This effect was not sensitive to cGMP-dependent protein kinase (PKG) inhibitors, whereas it was partially blocked by cAMP dependent protein kinase (PKA) inhibitors. The relaxation observed was also reverted up to 50+/-8.5% by iberiotoxin, a selective inhibitor of large conductance, calcium-activated potassium channels (BK(Ca)). Our results indicate that there exists a crosstalk mechanism between cAMP and cGMP signaling pathways which lead to relaxation of guinea pig tracheal smooth muscle and also that BK(Ca) channels are involved to a certain extent in this phenomenon. PMID- 14733950 TI - Mammalian mitochondrial and microsomal cytochromes b(5) exhibit divergent structural and biophysical characteristics. AB - The only outer mitochondrial membrane cytochrome b(5) examined to date, from rat (rOM b(5)), exhibits greater stability than known mammalian microsomal (Mc) isoforms, as well as a much higher kinetic barrier for hemin dissociation and a more negative reduction potential. A BlastP search of available databases using the protein sequence of rOM b(5) as template revealed entries for analogous proteins from human (hOM b(5)) and mouse (mOM b(5)). We prepared a synthetic gene coding for the heme-binding domain of hOM b(5), and expressed the protein to high levels. The hOM protein exhibits stability, hemin-binding, and redox properties similar to those of rOM b(5), suggesting that they are characteristic of the OM b(5) subfamily. The divergence in properties between the OM and Mc b(5) isoforms in mammals can be attributed, at least in part, to the presence of two extended hydrophobic patches in the former. The biophysical properties characteristic of the OM proteins may be important in facilitating the two functions proposed for them so far, reduction of ascorbate radical and stimulation of androgen synthesis. PMID- 14733951 TI - Substrate modulates compound I formation in peroxide shunt pathway of Pseudomonas putida cytochrome P450(cam). AB - The active oxygenating intermediate, a ferryl-oxo-(II) porphyrin cation radical (compound I), in substrate-bound cytochrome P450(cam) (P450(cam)) has eluded detection and kinetic analysis for several decades. Upon rapid mixing of peroxides-H(2)O(2) and m-CPBA with substrate-bound forms of P450(cam), we observed an intermediate with spectral features characteristic of compound I. Unlike in H(2)O(2), kinetic investigation on the reaction of m-CPBA with various substrate (camphor, adamantone, and norcamphor)-bound P450(cam) and its Y96A mutant shows a preferential binding of the aromatic end group of m-CPBA to the active-site of the enzyme and modulation of compound I formation by the local environment of heme active-site. The results presented in this paper describe the importance of heme environment in modulating formation of compound I, and form the first kinetic analysis of this intermediate in the peroxide shunt pathway of substrate-bound P450(cam). PMID- 14733952 TI - Effects of L- or D-Pro incorporation into hydrophobic or hydrophilic helix face of amphipathic alpha-helical model peptide on structure and cell selectivity. AB - A synthetic amphipathic alpha-helical model peptide, KLW, displays non-cell selective cytotoxicity. To investigate the effects of L- or D-Pro kink incorporation into hydrophobic or hydrophilic helix face of KLW on structure, cell selectivity, and membrane-binding affinity, we designed a series of four peptides, in which Leu(9) and Lys(11) in the hydrophobic and hydrophilic helix face of KLW, respectively, are substituted with L- or D-Pro. A L- or D-Pro substitution (KLW-L9P or KLW-L9p) of Leu(9) at the hydrophobic helix face of KLW induced a more significant reduction in hemolytic activity with improved antibacterial activity than that (KLW-K11P or KLW-K11p) of Lys(11) in the hydrophilic helix face. In addition, D-Pro-containing peptides (KLW-L9p and KLW K11p) displayed less hemolytic activity than L-Pro-containing peptides (KLW-L9P and KLW-K11P). Tryptophan fluorescence studies revealed that bacterial cell selectivity of KLW-L9P, KLW-L9p, and KLW-K11p is closely related to selective interactions with negatively charged phospholipids. CD analysis revealed that L- or D-Pro incorporation into KLW reduces the alpha-helicity of the peptide and D Pro incorporation induces more significant disruption in alpha-helical structure than L-Pro incorporation. Our results collectively suggest that D-Pro incorporation into the hydrophobic helix face of non-cell selective amphipathic alpha-helical peptides may be useful for the design of novel antimicrobial peptides possessing high bacterial cell selectivity without hemolytic activity. PMID- 14733953 TI - Phosphorylation of p68 RNA helicase regulates RNA binding by the C-terminal domain of the protein. AB - We previously reported ATPase, RNA unwinding, and RNA-binding activities of recombinant p68 RNA helicase that was expressed in Escherichia coli. Huang et al. The recombinant protein bound both single-stranded (ss) and double-stranded (ds) RNAs. To further characterize the substrate RNA binding by p68 RNA helicase, we expressed and purified the recombinant N-terminal and C-terminal domains of the protein. RNA-binding property and protein phosphorylation of the recombinant domains of p68 were analyzed. Our data demonstrated that the C-terminal domain of p68 RNA helicase bound ssRNA. More interestingly, the C-terminal domain was a target of protein kinase C (PKC). Phosphorylation of the C-terminal domain of p68 abolished its RNA binding. Based on our observations, we propose that the C terminal domain is an RNA substrate binding site for p68. The protein phosphorylation by PKC regulates the RNA binding of p68 RNA helicase, which consequently controls the enzymatic activities of the protein. PMID- 14733954 TI - Ectopic deposition of melanin pigments as detoxifying mechanism: a paradigm for basal nuclei pigmentation. AB - Melanins are UV shielding pigments found in skin and other light exposed tissues. However, a kind of melanin, named neuromelanin (NM), is found in those deep brain loci that degenerate in Parkinson's disease (PD), where no such a function may be imagined. The NM synthetic pathway, different from the one of eumelanin based on tyrosinase, is still obscure as well as its physiological function. Here we show that under conditions of excess of toxic quinone concentration, nonmelanocytic cell strains (i.e., primary keratinocytes) may accumulate a dark cytoplasmatic pigment that proved to be a melanin. The ability of pigment deposition, possibly driven by peroxidases, is restricted to diploid cells and increases cell survival acting as a sink for potentially hazardous quinones. We suggest that in the basal nuclei, exposed to high level of catecholaminergic neurotransmitters, NM deposition is a relevant antioxidant mechanism by trapping quinones and semiquinones, thus protecting neurons from accumulating damage over many years. In this perspective, just as a hypothesis, we may imagine that PD neuron degeneration is the consequence of a reduced/abrogated ability to produce neuromelanin. PMID- 14733955 TI - Two inhibitor molecules bound in the active site of Pseudomonas sedolisin: a model for the bi-product complex following cleavage of a peptide substrate. AB - High-resolution crystallographic analysis of a complex of the serine-carboxyl proteinase sedolisin with pseudo-iodotyrostatin revealed two molecules of this inhibitor bound in the active site of the enzyme, marking subsites from S3 to S3('). The mode of binding represents two products of the proteolytic reaction. Substrate specificity of sedolisin was investigated using peptide libraries and a new peptide substrate for sedolisin, MCA-Lys-Pro-Pro-Leu-Glu#Tyr-Arg-Leu-Gly Lys(DNP)-Gly, was synthesized based on the results of the enzymatic and crystallographic studies and was shown to be efficiently cleaved by the enzyme. The kinetic parameters for the substrate, measured by the increase in fluorescence upon relief of quenching, were: k(cat)=73+/-5 s(-1), K(m)=0.12+/ 0.011 microM, and k(cat)/K(m)=608+/-85 s(-1)microM(-1). PMID- 14733956 TI - Eotaxin and monocyte chemotactic protein-3 use different modes of action. AB - Eotaxin selectively binds CC chemokine receptor (CCR) 3, whereas monocyte chemotactic protein (MCP)-3 binds CCR1, CCR2, and CCR3. To identify the functional determinants of the chemokines, we generated four reciprocal chimeric chemokines-M10E9, M22E21, E8M11, and E20M23-by shuffling the N-terminus and N loop of eotaxin and MCP-3. M22E21 and E8M11, which shared the N-loop from MCP-3, bound to monocytes with high affinity, and activated monocytes. In contrast, M10E9 and E20M23, which lacked the N-loop, failed to bind and transduce monocyte responses, identifying the N-loop of MCP-3 as the selectivity determinant for CCR1/CCR2. A BIAcore assay with an N-terminal peptide of CCR3 (residues 1-35) revealed that all chimeras except E20M23 exhibited varying degrees of binding affinity with commensurate chemotaxis activity of eosinophils. Surprisingly, E20M23 could neither bind the CCR3 peptide nor activate eosinophils, despite having both N-terminal motifs from eotaxin. These results suggest that the two N terminal motifs of eotaxin must cooperate with other regions to successfully bind and activate CCR3. PMID- 14733957 TI - Expression of trypsin-like proteases and protease nexin-1 in mdx mouse muscles. AB - In order to examine the possible participation of trypsin-like proteases in the onset and progress of muscular dystrophy, we investigated the expression of the trypsin-like protease in muscular tissues in mdx mice. We found that the mRNAs of several trypsin-like proteases, including hepsin and t-PA, were expressed in the muscular tissues of mdx mice, but at levels not significantly different from normal mice. Since the enzymatic properties of dystrypsin, a muscle trypsin-like protease activated before onset of the disease, are similar to those of thrombin, we investigated the expression pattern of thrombin in mdx mouse muscles. The results showed that prothrombin mRNA is up-regulated in mdx mice at 20-30 days of age but not before the age of 15 days (preclinical). Since protease nexin-1 (PN 1) is known to be a physiological inhibitor of thrombin, we also examined the expression pattern of PN-1. We found that PN-1 transcription and translation is down-regulated in the muscular tissues of mdx mice, before the onset of clinical symptoms. These results suggest that thrombin may be involved in the progression of muscular dystrophy or the regeneration of muscle fibers after the onset of the disease and that the reduced level of PN-1 may enhance the activities stimulate the activities of muscle proteases, including dystrypsin, at a preclinical stage in mdx mice. PMID- 14733958 TI - Congenital form of spinal muscular atrophy predominantly affecting the lower limbs: a clinical and muscle MRI study. AB - We describe clinical and muscle magnetic resonance imaging (MRI) findings in 11 cases (three familial and eight sporadic) with the form of spinal muscular atrophy characterised by predominant involvement of the lower limbs with weakness of the proximal and distal muscles and marked atrophy of the distal leg and foot muscles. All patients presented at birth with talipes, which were in extension in seven of the 11. Arm muscle and function were preserved and lower limbs appeared to be disproportionately shorter compared to trunk and upper limbs. Functional abilities were markedly affected and only one of the 11 is able to walk independently for long distances, while six require support of crutches and two use callipers for walking. One child lost ambulation following a fall. The course of the disease is relatively stable and the progression of disability appeared to be related mostly to increased contractures rather than to loss of muscle strength. Respiratory and cardiac function were well preserved. A neurogenic disorder was suggested by electromyography and/or muscle biopsy in all patients, while motor nerve conduction was consistently normal. Muscle MRI of the thighs revealed diffuse atrophic appearance with relative hypertrophy of the adductor longus and of the semitendinosus. Genetic studies excluded the involvement of the survival motor neuron gene but none of these families was sufficiently informative to study linkage to the locus on chromosome 12q23-q24 previously found to be involved in patients with similar phenotype. In our experience this form of spinal muscular atrophy affecting predominantly the lower limbs is a relatively common form and should be considered in the differential diagnosis of infants with talipes and weakness in the lower limbs. The identical clinical and imaging features of the sporadic and familial cases suggest that these cases are likely to be affected by the same condition. PMID- 14733959 TI - Pilot trial of phenylbutyrate in spinal muscular atrophy. AB - The aim of this study was to evaluate tolerability and efficacy of phenylbutyrate (PB) in patients with spinal muscular atrophy (SMA). Ten patients with SMA type II confirmed by DNA studies (age range 2.6-12.7 years, mean age 6.01) were started on oral PB (triButyrate) in powder or tablets. The dosage was 500 mg/kg per day (maximum dose 19 g/d), divided in five doses (every 4 h, skipping one night-dose) using an intermittent schedule (7 days on and 7 days off). Measures of efficacy were the change in motor function from baseline to 3 and 9 weeks, by means of the Hammersmith functional motor scale. In children older than 5 years, muscle strength, assessed by myometry, and forced vital capacity were also measured. We found a significant increase in the scores of the Hammersmith functional scale between the baseline and both 3-weeks (P < 0.012) and 9-weeks assessments (P < 0.004). Our results indicate that PB might be beneficial to SMA patients without producing any major side effect. Larger prospective randomised, double-blind, placebo controlled trials are needed to confirm these preliminary findings. PMID- 14733960 TI - Cardiovascular autonomic control in myotonic dystrophy type 1: a correlative study with clinical and genetic data. AB - The autonomic nervous system has been evaluated in myotonic dystrophy with contradictory results and its relationship with heart disturbances remains unclear. Twenty-three patients with myotonic dystrophy type 1 were investigated by a battery of six cardiovascular autonomic tests and power spectral analysis of heart rate variability. Although 15 patients (65%) revealed abnormal or borderline results in some tests, only one patient had a definite autonomic damage, as indicated by two or more abnormal tests. As a group, myotonic dystrophy type 1 patients showed a significant reduction of heart rate variability during deep breathing (P < 0.0001). The exclusive involvement of parasympathetic tests suggests that a mild vagal dysfunction occurs in some myotonic dystrophy type 1 patients. The results indicate that such autonomic abnormalities are not: (1) part of a peripheral neuropathy; (2) related to cytosine-thymine-guanine repeat size or breathing pattern. Power spectral analysis showed a reduction of supine low-frequency band, which is, but not exclusively, a marker of sympathetic activity. It was inversely correlated to disease duration (P < 0.04), suggesting a progression as the disease advances. A low-frequency power, recorded after standing, was significantly associated (P < 0.02) with presence of heart involvement. Our findings suggest that a mixed, especially parasympathetic, autonomic dysfunction may occur in myotonic dystrophy type 1, although it is not a major finding. It could play a role in the occurrence of cardiac abnormalities, or increase the risk of sudden cardiovascular events. PMID- 14733961 TI - Increased resting energy expenditure in subjects with Emery-Dreifuss muscular dystrophy. AB - We have studied changes in energy expenditure and body composition in adult males with Emery-Dreifuss muscular dystrophy, age-matched males with hyperCKemia and age-matched healthy controls. All participants were studied twice, 2-3 years apart. Resting energy expenditure was studied by indirect calorimetry, lean body mass and body fat by dual X-ray absorptiometry, and muscle mass was estimated based on 24-h urinary creatinine excretion. At baseline and 2-3 years later, body fat was significantly higher (P < 0.011 and P < 0.003, respectively) and lean body mass significantly lower (P < 0.024 and P < 0.012, respectively) in patients with Emery-Dreifuss muscular dystrophy as compared to subjects with hyperCKemia and healthy controls. Resting energy expenditure, over the study period, increased significantly in patients with Emery-Dreifuss muscular dystrophy (P < 0.031), but not in patients with hyperCKemia nor in healthy controls. Our study suggests that patients with Emery-Dreifuss muscular dystrophy may have increased energy expenditure relative to healthy subjects. If not met by increased caloric intake, this greater energy expenditure may partially contribute to a further deterioration in their muscle performance. PMID- 14733962 TI - The novel neurofilament light (NEFL) mutation Glu397Lys is associated with a clinically and morphologically heterogeneous type of Charcot-Marie-Tooth neuropathy. AB - Charcot-Marie-Tooth disease comprises a heterogeneous group of hereditary neuropathies which fall into two main groups: demyelinating CMT1 with reduced nerve conduction velocity and axonal CMT2 with normal nerve conduction velocity. The neuropathological features correspond in most cases to this classification. Four genes were recently identified to cause autosomal dominant CMT2, including the neurofilament light gene. Thus far, only few mutations have been reported in neurofilament light involving eight amino acids of the gene. We identified a novel mutation, Glu397Lys, in a conserved motive signaling the end of the rod domain. The affected family members from three generations showed strikingly different clinical phenotypes, including weakness of the lower extremities, foot deformities, and deafness. The mutation was associated with nerve conduction velocities ranging from 27 m/s in a 25-year-old female to 43 m/s in an 82-year old male in the lower extremity motor nerves. Sural nerve biopsies of two affected subjects were analyzed by light and electron microscopy. The pathological changes consisted of a reduction of predominantly large myelinated nerve fibers and various stages of onion bulb formation as typically seen in CMT1. This correlative study further confirms that neurofilament light gene mutations cause a wide clinical spectrum. Thus, analysis of the neurofilament light gene should not be restricted to pure axonal neuropathies. PMID- 14733963 TI - A Japanese patient with distal myopathy with rimmed vacuoles: missense mutations in the epimerase domain of the UDP-N-acetylglucosamine 2-epimerase/N acetylmannosamine kinase (GNE) gene accompanied by hyposialylation of skeletal muscle glycoproteins. AB - Hereditary inclusion body myopathy and distal myopathy with rimmed vacuoles are both caused by mutations of the UDP-N-acetylglucosamine 2-epimerase/N acetylmannosamine kinase (GNE) gene. Here we report a Japanese patient with compound heterozygous missense mutations in the epimerase domain of GNE gene, 89 G to C and 578 A to T. Biochemical analysis demonstrated decreased reactivity of skeletal muscle glycoproteins with the lectins recognizing sialic acid residues. The results suggest that hyposialylation of glycoproteins may be involved in the pathogenesis of muscle dysfunction in this patient. PMID- 14733964 TI - A novel sporadic mutation in cytochrome c oxidase subunit II as a cause of rhabdomyolysis. AB - Disorders of the mitochondrial genome are an important cause of neurological disease, with patients presenting a variety of different phenotypes. Exercise induced muscle pain and myoglobinuria have been described with a number of metabolic defects, but because of the enormous variability of the mitochondrial genome identifying causative mitochondrial DNA mutations can be extremely difficult. Since mitochondrial tRNA genes were considered to be hot spots for mutation, sequencing was initially often confined to these genes. In a patient with symptoms and signs of exercise intolerance and myoglobinuria we originally ascribed pathogenicity to a mitochondrial-tRNA(Phe) mutation but here we show that the true pathogenic mutation was a novel mutation in the gene encoding subunit II of cytochrome c oxidase. We believe that this study demonstrates the importance of whole mitochondrial genome sequencing and of access to large sequence databases. PMID- 14733965 TI - Functional characterisation of a mutant actin (Met132Val) from a patient with nemaline myopathy. AB - The mutation Met132Val in the ACTA1 gene was identified in a patient with mild nemaline myopathy (NM). We examined actin mRNA and protein from biopsy samples. Sixty-one percent of the mRNA from the biopsy was not cleaved with BstX1, indicating the presence of mutant messenger in vivo. Monomeric actin was extracted from 2.5 mg of mutant muscle and wild type muscle. A proportion of the NM actin did not polymerise in 50 mM KCl, 2.5 mM MgCl2 but all the wild-type actin did. NM actin was fully polymerised by 50 mM KCl, 2.5 mM MgCl2, 150 nM rhodamine-phalloidin. Thin filaments reconstituted with this co-polymer were different from wild-type. The NM actin produces faster sliding of thin filaments at pCa5 and higher relative isometric force. We conclude that the mutant mRNA and protein is expressed and that the mutation reduces polymerisability and alters thin filament function. PMID- 14733966 TI - Macrophagic myofasciitis: an infantile Italian case. AB - Macrophagic myofasciitis is a recently identified inflammatory myopathy mostly described in adult French patients complaining of arthro-myalgias and fatigue. It is probably due to intramuscular injection of aluminium-containing vaccines and is characterized by a typical muscular infiltrate of large macrophages with aluminium inclusions. We report a 1-year-old Italian child presenting irritability, delayed motor development, hyperCKemia (up to 10 times the normal value), and typical features of macrophagic myofasciitis on muscle biopsy. The child recovered fully after steroid therapy. Macrophagic myofasciitis is a new treatable cause of motor retardation and hyperCKemia in children, and is probably more common than reported. Diagnosis requires a high index of suspicion and can be missed if biopsy is performed outside the vaccination site. PMID- 14733972 TI - Weight loss in adults with Down syndrome and with dementia in Alzheimer's disease. AB - An association between weight loss and Alzheimer's disease has been established in the general population but little information is available regarding this association in people with intellectual disabilities. A 4-year longitudinal study of adults with Down syndrome with and without Alzheimer's disease was undertaken. Age-associated weight loss was seen in virtually all older adults with Down syndrome. A significant association between weight loss and Alzheimer's disease was found for older adults with Down syndrome. This study highlights important research and clinical issues regarding weight loss and nutrition in Down syndrome adults with dementia. PMID- 14733973 TI - Weight survey on adult persons with mental retardation living in the community. AB - Prevalence of underweight and obesity were investigated in 282 mentally retarded persons living on the West Coast of Norway. Data collected in this survey suggest that people with severe mental retardation were more likely to be underweight and people with mild mental retardation were more likely to be obese. Compared to persons of average intellectual levels, persons with mental retardation were found to be over-represented with both underweight and obesity. Food refusal and self-induced vomiting was more present among persons regarded as underweight and reduced control of food intake was related to obesity. Findings suggest that behavioral variables may have explanatory value with respect to both underweight and obesity. Further investigation using functional analysis of behavior is recommended. PMID- 14733974 TI - Relevance of functional behavioral assessment research for school-based interventions and positive behavioral support. AB - The Individuals With Disabilities Education Act (IDEA) Amendments of 1997 mandate the use of functional behavioral assessment (FBA) and positive behavioral supports and interventions for students with disabilities. Although much progress has been made in our understanding of functional analysis over the past 15 years, the extent to which these findings can be generalized across clients, methods, settings, and response classes is unknown. This article reviewed 150 school-based intervention studies conducted with children published in the Journal of Applied Behavior Analysis from 1991 to 1999. Fifty-two percent of these intervention studies did not report using FBA information linked to the intervention. Interventions based on descriptive, experimental, or combined FBA procedures were no more effective than interventions in which no FBA information was provided. With respect to positive behavioral support, over half of the studies targeted appropriate behaviors with two-thirds of them using a combination of antecedent- and consequence-based interventions. Recommendations are made for conducting fundamental research on reliability and validity issues in FBA and determining when, how, and under what conditions FBA procedures are most appropriate. PMID- 14733975 TI - Effects of familiar versus unfamiliar therapists on responding in the analog functional analysis. AB - The analog functional analysis involves the manipulation of pre-determined antecedent and consequent events and typically is conducted by trained experimenters. Inclusion of idiosyncratic variables in the analog functional analysis may affect responding. Inclusion of caregivers is one potential antecedent that may affect problem behavior. The purpose of this study was to evaluate the effects of including caregivers in the analog. Four individuals with developmental disabilities and their caregivers served as participants. For 3 of 4 participants, different patterns of responding were observed when caregivers versus experimenters conducted the functional analysis. PMID- 14733976 TI - Sleep problems as possible predictors of intensified symptoms of autism. AB - Researchers have been placing an increased importance on discovering what variables contribute to better prognosis during behavioral interventions for children with autism. This article preliminarily identifies sleep problems that may exacerbate symptoms of autism; thus, possibly influencing effectiveness of daytime interventions. A data-base of parent report of sleep problems of children with autism (N=55), ranging from 5 to 12 years of age (M=8.2 years) was evaluated. Results suggested that fewer hours of sleep per night predicted overall autism scores and social skills deficits. Similarly, stereotypic behavior was predicted by fewer hours of sleep per night and screaming during the night. Increased sensitivity to environmental stimuli in the bedroom and screaming at night predicted communication problems. Finally, sensitivity to environmental stimuli in the bedroom also predicted fewer developmental sequence disturbances. The results indicate that sleep problems and the diagnostic characteristics of autism may be related. However, future research must be completed to determine the specific relationship. PMID- 14733977 TI - An investigation of instructional scheduling arrangements for community-based instruction. AB - The instructional scheduling arrangements of simulated and community-based instruction across an equivalent set of functional and vocational skills were examined. Five secondary age students with moderate intellectual disabilities participated in four instructional scheduling arrangements measuring skill acquisition, generalization, and maintenance of functional tasks. The four instructional scheduling arrangements examined were simulated-only instruction (SOI), community-based instruction only (CBI), combination of SOI and CBI on consecutive school days (CCD), and combination of SOI and CBI on the same school day (CSD). The CSD schedule was significantly more effective for student acquisition performance than SOI, CBI, and CCD schedules. Although the CBI schedule resulted in the fewest number of instructional sessions for students to acquire the targeted skill, fewer sessions for skill generalization were required for students during the combined simulation and community instruction CSD schedule. Overall, both combined instructional scheduling arrangements (e.g., CCD, CSD) produced more efficient outcomes for generalization than SOI and CBI scheduling arrangements. PMID- 14733978 TI - An analysis of Snoezelen equipment to reinforce persons with severe or profound mental retardation. AB - Systematically developing methods of reinforcement for persons with severe and profound mental retardation has only recently received a good deal of attention. This topic is important since professionals in the field often have difficulty identifying sufficient numbers of positive stimuli. Snoezelen equipment as reinforcement for individuals with severe and profound mental retardation was evaluated because of the promise it holds for this population. Types of Snoezelen equipment which were most often approached and most reinforcing for these persons were identified. Implications of the findings for future assessment and treatment are discussed. PMID- 14733980 TI - Bridging the gap: bringing together intentional and unintentional injury prevention efforts to improve health and well being. AB - PROBLEM: Intentional and unintentional injury prevention efforts have traditionally been independent and non-integrated. Fostering collaboration between the sub-fields would enhance work within both sub-fields and advance injury prevention work as a whole. METHOD: A systematic assessment of similarities and differences between the sub-fields was performed, including an examination of relevant definitions and norms, research methods and findings, key risk and resiliency factors, and prevention strategies that would promote collaboration and better advance current prevention efforts. RESULTS/SUMMARY: Several areas exist in which injury prevention efforts could be coordinated or ideas and practices could be cross-applied, including training of practitioners, data collection and analysis, application of tools and methodologies, examination of risk and resiliency factors, and identification of funding sources and partners. IMPACT ON INDUSTRY: This paper delineates how intentional and unintentional injury prevention practitioners can more effectively collaborate to promote safer environments and further reduce incidence of injury. An integrated injury prevention approach could significantly impact the underlying contributors to both types of injury, allowing practitioners within both sub-fields to achieve greater outcomes through increased credibility, reduced duplication of efforts, more efficient use of resources, and unified injury prevention messages. PMID- 14733981 TI - Analysis of motor-vehicle crashes at stop signs in four US cities. AB - PROBLEM: Nearly 700000 police-reported motor vehicle crashes occur annually at stop signs, and approximately one-third of these crashes involve injuries. The purpose of this study was to develop a better understanding of the crashes that occur at stop signs and to identify potential countermeasures. METHOD: Police reports of crashes at stop sign-controlled intersections during 1996-2000 in four U.S. cities were examined in detail. At total of 1788 crash reports for intersections with two-way stop signs were included in the study. RESULTS: Stop sign violations accounted for about 70% of all crashes. Typically these crashes were angular collisions. Among crashes not involving stop violations, rear-end crashes were most common, accounting for about 12% of all crashes. Stop sign violation crashes were classified into several subtypes - driver stopped, driver did not stop, snow/wet/ice, and other/unknown. In about two-thirds of stop sign violation crashes, drivers said they had first come to a stop. In these cases, inability or failure to see approaching traffic often was cited as the cause of the crash. Drivers younger than 18 as well as drivers 65 and older were disproportionately found to be at fault in crashes at stop signs. IMPACT ON INDUSTRY: Potential countermeasures include changing traffic control and intersection design, improving intersection sight distance, and increasing conspicuity of stop signs through supplemental pavement markings and other devices. PMID- 14733982 TI - Views of US drivers about driving safety. AB - PROBLEM: To assess how drivers view dangers on the highway, what motivates them to drive safely, how they say they reduce their crash and injury risk, and how they rate their own driving skills. RESULTS: Most drivers rated their skills as better than average. The biggest motivating factor for safe driving was concern for safety of others in their vehicle, followed by negative outcomes such as being in a crash, increased insurance costs, and fines. The greatest threats to their safety were thought to be other drivers' actions that increase crash risk such as alcohol impairment or running red lights. In terms of reducing crashes and injuries, drivers tended to focus on actions they could take such as driving defensively or using seat belts. There was less recognition of the role of vehicles and vehicle features in crash or injury prevention. IMPACT ON RESEARCH, PRACTICE, AND POLICY: Knowing how drivers view themselves and others, their concerns, and their motivations and techniques for staying out of trouble on the roads provides insight into the difficulty of changing driving practices. PMID- 14733983 TI - Comprehension and memory for warning symbols: age-related differences and impact of training. AB - PROBLEM: Prior research indicates that many warning symbols are poorly understood, particularly by the elderly. METHOD: The effectiveness of three different training conditions to improve comprehension and memory for warning symbols was assessed for younger (18-35 years of age) and older (50-67 years of age) participants. All three conditions paired the symbols with associated text during training; however they differed in the extent to which they further elaborated on the meanings of the symbols. RESULTS: Training substantially improved accuracy and speed of responding on a comprehension test; however there was little difference among conditions. Additionally, while the magnitude of the training effect was similar for both age groups, older participants performed much more poorly than younger participants, both before (37% vs. 52% correct) and after training (68% vs. 88% correct on the immediate post-test), and found it more difficult to reject incorrect meanings (55% vs. 68% correct). CONCLUSION: Relatively simple training conditions can dramatically improve accuracy and speed of responding to warning symbols. IMPACT ON INDUSTRY: Training should be used to improve warning symbol comprehension since failures to adequately understand warning information may lead to injury or death. Furthermore, attempts should be made to address individual differences in warning processing such as those related to changes in cognitive processing across the lifespan. PMID- 14733984 TI - Deaths in construction related to personnel lifts, 1992-1999. AB - PROBLEM: This study examined deaths of construction workers due to personnel lifts (boom-supported and scissor lifts, suspended scaffolds, and crane platforms). METHODS: Deaths of construction workers for 1992-1999 were examined using data from the Census of Fatal Occupational Injuries, a Bureau of Labor Statistics database. RESULTS: The study identified 339 deaths: 42% from boom supported lifts; 26% from suspended scaffolds; 19% from scissor lifts; 5% from crane platforms; and 7% from unapproved lifts (e.g., forklift platforms). The main causes of death were falls (36%), collapses/tipovers (29%), and electrocutions (21%). DISCUSSION: Recommendations include: following OSHA regulations, wearing personal fall protection equipment, adequate maintenance, inspection before use, and training on the model of lift used. Precautions are also needed to prevent contact with overhead power lines. IMPACT ON INDUSTRY: The increasing popularity of boom-supported lifts and scissor lifts, both in construction and other industries, make their safety an important issue. PMID- 14733985 TI - The concept of degraded images applied to hazard recognition training in mining for reduction of lost-time injuries. AB - INTRODUCTION: This paper discusses the application of a training intervention that uses degraded images for improving the hazard recognition skills of miners. METHOD: NIOSH researchers, in an extensive literature review, identified fundamental psychological principles on perception that may be employed to enhance the ability of miners to recognize and respond to hazards in their dangerous work environment. Three studies were conducted to evaluate the effectiveness of the degraded image training intervention. A model of hazard recognition was developed to guide the study. RESULTS: In the first study, miners from Pennsylvania, West Virginia and Alabama, who were taught with the aid of degraded images, scored significantly better on follow-up hazard recognition performance measures than those trained using traditional instructional methodologies. The second and third studies investigated the effectiveness of the training intervention at two mining companies. Data collected over a 3-year period showed that lost-time injuries at mines in Alabama and Illinois declined soon after the training intervention was instituted. IMPACT ON INDUSTRY: Further exploration of the hazard recognition model and the development of other interventions based on the model could support the validity of the steps in the hazard recognition model. PMID- 14733986 TI - Responsibility of drivers, by age and gender, for motor-vehicle crash deaths. AB - PROBLEM: Motor-vehicle crash rate comparisons by age and gender usually are based on the extent to which drivers in a particular age/gender category are themselves injured or involved in crashes (e.g., the number of 20-year-old females in crashes). Basing comparisons instead on the extent to which drivers in various age/gender groups are responsible for deaths (including themselves) in their crashes is more revealing of their overall contribution to the problem. METHODS: Data from the Fatality Analysis Reporting System (FARS, 1996-2000) were used in the analysis, which was based on crashes that involved one or two vehicles only. Drivers in fatal single-vehicle crashes were assumed to have responsibility for the crash. In fatal two-vehicle crashes, driver operator errors reported by police were used to assign crash responsibility. RESULTS: When all crashes were considered, both the youngest and oldest drivers were most likely to be responsible for deaths in their crashes. In two-vehicle crashes, the oldest drivers were more likely than young drivers to be responsible. Young males were more likely than young females to be responsible for crash deaths, whereas females in their 50s and older were more likely than same-age males to be responsible. In terms of responsibility for deaths per licensed driver, young drivers, especially males, had the highest rates because of their high involvement rates and high responsibility rates. The majority of deaths for which young drivers were responsible occurred to people other than themselves, especially passengers in their vehicles, whereas the bulk of the deaths for which older drivers were responsible were their own. DISCUSSION: The results highlight the contribution of young drivers to the motor-vehicle crash problem, the need for measures such as passenger restrictions in graduated licensing systems, and the need for vehicle modifications to better protect older occupants. PMID- 14733987 TI - Causal attributions of Ghanaian industrial workers for accident occurrence: miners and non-miners perspective. AB - PROBLEM: Reports from the accident literature indicate that accident rates tend to vary with type of occupation. The mining industry has been recorded as the most dangerous with a high disabling injury rate. This observation has been attributed to the extremely stressful conditions under which miners work. Besides, the intimidating work environment in the mines has been insinuated to invoke a sense of helplessness, fatalism and hence defensive causal attributions for accident occurrences. METHOD: This study compared causal attributions between accident victims in Ghana's mining industry with their counterparts in textile factories. T values and Chi-square were employed to test for statistically significant differences between the two groups of accident victims. RESULTS: Findings indicate that there is no difference between the causal attributions for miners and non-miners. IMPACT ON INDUSTRY: Accident frequency and occupational type have no impact on causal attributions. PMID- 14733988 TI - Differences in relative risks for fatal occupational highway transportation accidents. AB - PROBLEM AND METHOD: This study examined the trends in the relative risks for being involved in fatal occupational highway transportation accidents based upon the age and gender of the victim. RESULTS: Significant differences in relative risks were identified based upon age; however, there were no significant differences in relative risks based upon gender. The trend analysis of relative risks for all motor-vehicle accidents based upon age showed that males exhibited a significant cubic trend while females exhibited a significant linear trend. The trend analysis of relative risks for fatal motor-vehicle accidents specifically involving vehicle operators based upon age yielded a significant quadratic trend for males and no significant trends for females. Examining the relative risks for fatalities involving only motor-vehicle operators controlled, to a certain extent, the differences in job exposure to motor-vehicle accidents based upon age. IMPACT: Prevention measures are identified as most crucial for older male workers in the transportation and agriculture industries. PMID- 14733989 TI - Evaluation of a safety training program in three food service companies. AB - PROBLEM: Outcome measures for safety training effectiveness research often do not include measures such as occupational injury experience. Effectiveness mediators also receive sparse attention. METHOD: A new safety training curriculum was delivered to workers in a stratified random sample of food service facilities across three companies. A similar group of facilities received usual training. We collected post-test measures of demographic variables, safety knowledge, perceptions of transfer of training climate, and workers' compensation claim data for one year after the initial training activities. RESULTS: Knowledge test scores were apparently higher in the new-training units than in the usual training units. Some demographic variables were inconsistently associated with these differences. Evidence for reduction of the injury rate associated with the new training was observed from two companies but only approached significance for one company. A second company revealed a similar but non-significant trend. Knowledge scores were not significantly associated with lower injury rates. DISCUSSION: We found evidence that safety training increases knowledge and reduces injuries. We found almost no evidence of effects of training effectiveness mediators, including no relationship between safety knowledge and injury experience. Methodological issues related to conducting a large study may have influenced these results. IMPACT ON INDUSTRY: Although safety training leads to greater knowledge and, in some cases, reduced occupational injuries, the influence of mediating variables remains to be fully explained. PMID- 14733990 TI - Risky, aggressive, or emotional driving: addressing the need for consistent communication in research. AB - PROBLEM: Researchers agree that a consistent definition for aggressive driving is lacking. Such definitional ambiguity in the literature impedes the accumulation of accurate and precise information, and prevents researchers from communicating clearly about findings and implications for future research directions. This dramatically slows progress in understanding the causes and maintenance factors of aggressive driving. SUMMARY: This article critiques prevailing definitions of driver aggression and generates a definition that, if used consistently, can improve the utility of future research. Pertinent driving behaviors have been variably labeled in the literature as risky, aggressive, or road rage. The authors suggest that the term "road rage" be eliminated from research because it has been used inconsistently and has little probability of being clarified and applied consistently. Instead, driving behaviors that endanger or have the potential to endanger others should be considered as lying on a behavioral spectrum of dangerous driving. Three dimensions of dangerous driving are delineated: (a). intentional acts of aggression toward others, (b). negative emotions experienced while driving, and (c). risk-taking. IMPACT ON INDUSTRY: The adoption of a standardized definition for aggressive driving should spark researchers to use more explicit operational definitions that are consistent with theoretical foundations. The use of consistent and unambiguous operational definitions will increase the precision of measurement in research and enhance authors' ability to communicate clearly about findings and conclusions. As this occurs over time, industry will reap benefits from more carefully conducted research. Such benefits may include the development of more valid and reliable means of selecting safe professional drivers, conducting accurate risk assessments, and creating preventative and remedial dangerous driving safety programs. PMID- 14733991 TI - The use of supervisory practices as leverage to improve safety behavior: a cross level intervention model. AB - INTRODUCTION: The paper presents three intervention studies designed to modify supervisory monitoring and rewarding of subordinates' safety performance. METHOD: Line supervisors received weekly feedback concerning the frequency of their safety-oriented interactions with subordinates, and used this to self-monitor progress toward designated improvement goals. Managers higher up in the organizational hierarchy received the same information, coupled with synchronous data concerning the frequency of workers' safety behaviors, and highlighting co variation of supervisory action and workers' behavior. RESULTS: In all the companies involved, supervisory safety-oriented interaction increased significantly, resulting in significant changes in workers' safety behavior and safety climate scores. Continued improvement during the post-intervention period suggests the inclusion of workers' safety behavior as in-role supervisory responsibility. Applied and theoretical implications are discussed. PMID- 14733992 TI - Comparison of young male and female drivers' attitude and self-reported traffic behaviour in Finland in 1978 and 2001. AB - PROBLEM: This study evaluates how the traffic behaviors of young drivers and their attitudes toward traffic regulations have changed over the last 23 years, and particularly, whether the differences in attitudes and behavior between male and female drivers have changed. METHOD: The study was conducted in 2001, and it replicated a traffic attitude survey administered in 1978. The same survey was used, enabling comparison between the years. The number of respondents was 3158 in 1978 and 2759 in 2001. RESULTS: The comparison revealed several differences regarding the background factors, attitudes, and driving style of novice drivers. Most obvious changes in the drivers' background were the changes in education level (higher today), driver training (more private training today), and exposure/experience in terms of kilometers (more today). The summary variable measured that the young drivers showed more negative attitudes toward traffic rules and safe driving in 2001 compared to 1978. Female drivers drove less than males and evaluated their driving skill lower. Female drivers were less involved in accidents and they committed less traffic offenses than males (kilometrage controlled). Female drivers showed a more positive attitude toward traffic safety and rules than males. The difference in traffic attitudes and behavior between males and females in 1978 compared to 2001 remained the same or even increased somewhat. PMID- 14733993 TI - A comprehensive framework for assessing and selecting appropriate scaffolding based on analytic hierarchy process. AB - PROBLEM: Bamboo scaffolding has been widely used in South China, Hong Kong, and Southeast Asia for many years. Bamboo scaffolding is more economical, but its safety record is relatively poor. Conversely, metal scaffolding is more expensive, but its use in these regions has increased in recent years because it is relatively safe. The assessment and selection of the most appropriate type of scaffolding for a particular construction project is always a concern for project managers. METHOD: This paper suggests a comprehensive assessment framework that enables project managers to take a number of major quantitative and qualitative factors into consideration when making scaffolding decisions. This framework is based on the Analytic Hierarchy Process methodology and a survey among project managers in Hong Kong. RESULTS: The research reveals that the overall performance of metal scaffolding is believed to be better than bamboo scaffolding. A sensitivity analysis has also been conducted to investigate the impact of various factors on the final decisions. IMPACT ON INDUSTRY: The proposed assessment framework can be used as a supporting tool for project managers in the selection of scaffolding for their projects. PMID- 14733994 TI - Analysis of driver injury severity levels at multiple locations using ordered probit models. AB - PROBLEM: The occurrence and outcome of traffic crashes have long been recognized as complex events involving interactions between many factors, including the roadway, driver, traffic characteristics, and the environment. This study is concerned with the outcome of the crash. METHOD: Driver injury severity levels are analyzed using the ordered probit modeling methodology. Models were developed for roadway sections, signalized intersections, and toll plazas in Central Florida. All models showed the significance of driver's age, gender, seat belt use, point of impact, speed, and vehicle type on the injury severity level. Other variables were found significant only in specific cases. RESULTS: A driver's violation was significant in the case of signalized intersections. Alcohol, lighting conditions, and the existence of a horizontal curve affected the likelihood of injuries in the roadway sections' model. A variable specific to toll plazas, vehicles equipped with Electronic Toll Collection (ETC), had a positive effect on the probability of higher injury severity at toll plazas. Other variables that entered into some of the models were weather condition, area type, and some interaction factors. This study illustrates the similarities and the differences in the factors that affect injury severity between different locations. PMID- 14733995 TI - Current and future strategies for preventing and managing erectile dysfunction following radical prostatectomy. AB - INTRODUCTION AND OBJECTIVES: As radical prostatectomy remains a commonly used procedure in the treatment of clinically localized prostate cancer, we critically analyzed current and future strategies for preventing and managing postoperative erectile dysfunction. METHODS: Systematic literature review using Medline and CancerLit from January 1997 to June 2003. Abstracts published in the journals European Urology, The Journal of Urology and the International Journal of Impotence Research as official proceedings of internationally known scientific societies held in the same time period were also assessed. RESULTS: Patient selection and surgical technique are the major determinants of postoperative erectile function. Apoptosis of corporeal smooth muscle cells plays a role in the development of cavernous veno-occlusive dysfunction following radical prostatectomy. Pharmacological prophylaxis and treatment of postoperative erectile dysfunction is effective and safe. The concepts of cavernous nerve reconstruction and neuroprotection have been associated to promising results. CONCLUSIONS: In the hands of experienced surgeons, properly selected patients undergoing a nerve sparing radical prostatectomy should achieve unassisted or medically assisted erections postoperatively. PMID- 14733996 TI - Quality of life following treatment for early prostate cancer: does low dose rate (LDR) brachytherapy offer a better outcome? A review. AB - OBJECTIVE: Due to a lack of evidence from randomised studies, there is little agreement on the best form of treatment among men who require curative treatment for prostate cancer. The relative impact of the various treatments on symptoms and health-related quality of life is also controversial. We review the literature on quality of life changes following low dose rate brachytherapy (BXT) and compare BXT to other treatments for early prostate cancer. METHODS: Systematic literature review 1988-2003 (Medline). KEYWORDS: Brachytherapy; Radical prostatectomy; External beam radiotherapy; Quality of life; Symptoms. RESULTS: Review of the current literature suggests that radical prostatectomy, external beam radiotherapy and BXT either alone or in combination with supplementary external beam radiotherapy offer good long-term health-related quality of life. However differences exist in the toxicity of treatment in terms of erectile function, voiding difficulty, incontinence and bowel function. These differences seem to persist for at least 3-5 years post-treatment though longer term quality of life outcomes from modern techniques are unknown. CONCLUSION: BXT offers a high probability of maintaining continence, potency and normal rectal function though both storage and voiding urinary symptoms have been reported. Addition of androgen deprivation and EBRT to BXT may increase urinary, bowel and sexual toxicity of treatment. Quality of life outcome following brachytherapy compares favourably with other radical treatment options for the management of early prostate cancer. PMID- 14733997 TI - Bladder carcinoma in situ in 2003: state of the art. AB - Carcinoma is situ (CIS) of the bladder is a high-grade non-invasive malignancy with a high tendency of progression and transitional cell carcinoma outside the bladder. The diagnosis is a combination of abnormal cytology and cystoscopy with biopsies. Although cytology has clear limitations in low-grade lesions, such as a low inter- and intra-observer reproducibility, high-grade lesions and CIS should be diagnosed with a high degree of sensitivity and specificity. Currently available urinary markers do not (yet) seem to match cytology. The cystoscopic diagnosis is more difficult, since flat lesions are often difficult to see. The application of fluorescence cystoscopy and resection clearly improves the detection of the number of CIS lesions per patient and also the number of patients with CIS. For treatment of CIS (maintenance) BCG remains the golden standard. BCG appears to be able to prevent or delay progression to muscle invasive disease. BCG refractory patients are at high risk for progression and cancer death, and cystectomy is the treatment of choice. Alternatives for BCG refractory CIS patients, like intravesical chemo-immunotherapy, new chemotherapeutic drugs or photo-dynamic therapy, remain highly experimental. Last but not least, the danger for CIS patients is failure to respond to therapy and a high subsequent chance of progression and cancer-specific death. Unfortunately, despite much research, this prediction is not yet possible with molecular markers in daily practice. PMID- 14733998 TI - Surgical management of infiltrating bladder cancer in elderly patients. AB - OBJECTIVES: To review the surgical therapeutic options in elderly patients with infiltrating bladder cancer. METHODS: A review of the literature relevant to cystectomy and transurethral resection for infiltrating bladder cancer in elderly patients was conducted using Medline Services. RESULTS: Thanks to progress in anaesthesia, intensive care and surgery, cystectomy now forms part of the classical treatments for bladder cancer in elderly patients, with acceptable mortality and morbidity rates. The recent series of cystectomies performed in patients over 75 years old report a mortality rate associated with the procedure of less than 4.5%. The global mortality rate in the same population ranges from 10 to 50%. These rates are now similar to those reported in the general population. The mean survival after cystectomy in patients over 75 years old is more than 2 years. Global survival at 5 years is between 37 and 68%. It is acknowledged by most authors that resection alone is associated with higher relapse and progression rates than cystectomy. CONCLUSIONS: Cystectomy appears to be reasonable in elderly people who have a life expectancy of more than 2 years, provided that a rigorous pre-operative assessment and anaesthetic management are performed. Transurethral resection alone should be proposed only to patients with poor health status and/or very advanced age. PMID- 14733999 TI - Prostate cancer detection is also relevant in low prostate specific antigen ranges. AB - OBJECTIVE: To address detection rates and clinical features of the cancers detected with low prostate specific antigen (PSA) levels. METHODS: In the context of a prostate cancer (PCa) screening program 1097 men attended to a new rescreen round. Sextant prostate biopsy was recommended when PSA > or =3 ng/ml. We also recommended to undergo biopsy in the range 1.0-2.99 ng/ml when free to total (f/t) PSA ratio < or =20%. Detection rate was calculated and clinical features of the cancers detected were studied. RESULTS: Mean age was 61.1 years. A total of 497 (45.3%) had total PSA <1.0 ng/ml, 439 (40%) between 1.0 and 2.99 ng/ml, and 161 (14.7%) > or =3.0 ng/ml. In the group with PSA between 1.0 and 2.99 ng/ml and f/t PSA ratio < or =20% a total of 249 biopsies were indicated (159 performed, acceptance 63.9%), and 15 cancers detected (detection rate 9.4%). Biopsy was recommended to 72 men with PSA between 3.0 and 3.99 ng/ml, performed in 56 (77.8%), and 12 tumors detected (detection rate 21.4%). All cancers in the study were clinically localized. Only 4 out of 15 cancers with PSA in the range 1.0 2.99 ng/ml (26.7%) fulfilled clinical criteria of insignificant cancer. Two were poorly differentiated and found to have patologically extracapsular disease. None of the 12 patients with PCa and PSA between 3.0 and 3.99 ng/ml had poorly differentiated features and only one complied with criteria of insignificant cancer. One out of seven who underwent RRP was found to have extracapsular disease. CONCLUSIONS: Cancer detection in low PSA ranges is lower but still relevant. The detection of potentially aggresive cancers is still of concern. PMID- 14734000 TI - Cumulative prostate cancer risk assessment with the aid of the free-to-total prostate specific antigen ratio. AB - OBJECTIVE: To evaluate the cumulative risk of having a prostate cancer diagnosis in a repeated screening situation in relation to the free-to-total prostate specific antigen ratio (F/T-PSA). PATIENTS AND METHODS: The present study includes 1385 men (aged 50-70 years) who underwent prostate biopsy for the first time in the screening program that started in 1995. In case of a benign finding, the men have been followed biennially and new biopsies performed in case of persistently elevated PSA. The cumulative risk to be diagnosed with prostate cancer until July 1, 2002 was calculated by the Kaplan-Meier method and comparison was made between different levels of T-PSA and F/T-PSA ratios. RESULTS: Of 2129 biopsies 469 showed cancer. The cumulative 5-year risk to be diagnosed with prostate cancer was significantly dependent of the F/T-ratio. The risk for men with a T-PSA of 3-5.99 g/ml was 16% [6-25%] for those who had a ratio of >30% and 44% [34-60%] for those with a ratio of <10%. The corresponding difference for patients with a T-PSA of 6-9.99 g/ml was even more pronounced: 21% [0-42%] vs. 80% [64-96%]. CONCLUSION: By completing the T-PSA measurement with the F/T-PSA ratio it is possible to significantly better assess the cumulative prostate cancer risk within the next five years (without the aid of further urological work-up). PMID- 14734001 TI - Primary and pure neuroendocrine tumor of the prostate. AB - Primary neuroendocrine tumors of the prostate are very rare and their biologic behaviour is not yet well known. Clinical and histopathologic features of two cases, one with lymph-node involvement and one organ-confined are described. Young age, clinical presentation and good outcome after radical retropubic prostatectomy were comparable in both patients. PMID- 14734002 TI - A vaccination strategy for the long-term suppression of androgens in advanced prostate cancer. AB - OBJECTIVES: We have previously reported the ability of D17DT (formerly GnRH-DT) vaccination to produce castrate levels of androgens in men with advanced prostate cancer. This study examines the efficacy and tolerability of 3 and 15 micrograms of D17DT in 12 patients with advanced prostate cancer to establish a dose response relationship. METHODS: 12 patients received either 3 or 15 micrograms of D17DT as 3 deep intramuscular injections over 6 weeks. Outcome was assessed in terms of physical and biochemical evaluations of clinical progression and antibody titres. RESULTS: Significant titres of anti-GnRH antibodies were detected in 2 out of 6 subjects who received 15 micrograms of D17DT; suppression of testosterone to castrate levels accompanied by a significant and prolonged reduction in PSA was also demonstrated. No responses were seen following treatment with 3 micrograms of D17DT. CONCLUSION: The induction of anti-GnRH antibodies through vaccination with 15 micrograms D17DT can produce and sustain castrate levels of testosterone in men with advanced prostate cancer. PMID- 14734003 TI - Clinical outcome of a large-scale multi-institutional retrospective study for locally advanced bladder cancer: a survey including 1131 patients treated during 1990-2000 in Japan. AB - OBJECTIVE: We conducted a multi-institutional analysis to establish the contemporary clinical outcome of invasive bladder cancer treated with radical cystectomy in Japan. METHODS: A total of 1131 consecutive patients who underwent radical cystectomy for invasive bladder cancer between January 1990 and December 2000 at 32 hospitals were retrospectively analyzed. RESULTS: Histopathological analysis demonstrated that 1042 patients (92.1%) harbored transitional cell carcinomas (TCCs), whereas 89 patients (7.9%) presented non-TCCs, including squamous cell carcinoma and adenocarcinoma. Pelvic lymphadenectomy was performed in 1013 patients in total, and pathologically confirmed lymph node metastases were found in 162 (16.0%). The overall survival at 5 years was 68.0% and most deaths (79.0%) occurred within 3 years. Multivariate analysis demonstrated that gender, clinical stage, pathological stage, lymph node involvement and lymph node dissection were the independent predictive factors for survival, whereas histological type, sex and grade had no significant impact on survival. CONCLUSIONS: These clinical results demonstrate that radical cystectomy with lymph node dissection results in good survival for invasive bladder cancer, providing standard data with which other forms of therapy can be compared. PMID- 14734004 TI - Intravesical gemcitabine: a phase 1 and pharmacokinetic study. AB - INTRODUCTION: Superficial bladder cancer can be treated by transurethral resection and additional intravesical therapy. Although agents like Mitomycin C, Epirubicin and BCG are routinely used, there is a need for more potent and/or less toxic agents. Gemcitabine is a deoxycytidine analogue, used systemically for several tumours, such as non-localised bladder cancer, where it is effective and well tolerated. We investigated the use of three dose levels of gemcitabine when given intravesically in humans for safety and pharmacokinetic research. MATERIAL AND METHODS: Patients with superficial bladder cancer, except pT1G3 or CIS were included. Six weekly instillations of 1000, 1500 or 2000 mg gemcitabine were given in 50 ml saline for one hour. Dose modifications were defined in case of dose limiting toxicities. Blood samples were taken before, and 5, 30, 60 (= evacuation) and 120 minutes after instillations 1, 3 and 6. Samples were used for blood counts and pharmacokinetics. Side effects were noted. RESULTS: 3, 4 and 3 patients were treated with 1000, 1500, and 2000 mg gemcitabine respectively, of which 2, 3 and 1 patients had highly recurrent tumours before treatment. Seven patients experienced side effects: 2 with dysuria after the first instillation, 2 after instillations 3-6 and 4-6 and in 3 patients headache, fatigue and heavy legs were experienced once. All side effects were reversible, non-limiting and WHO 1. No macroscopic hematuria was seen. Haematology showed only one case of drop in white blood cell count (lowest dose level, after the first instillation). Gemcitabine plasma levels were immeasurable or low, with peak levels between 30 and 60 minutes, decreasing after more instillations. The metabolite difluorodeoxyuridine reached levels of at most 5 microM, indicating a very low passage of the drug to the systemic circulation. CONCLUSION: Intravesical gemcitabine in the dose used has minimal and reversible side effects. Plasma evaluation indicates that its intravesical use is safe. PMID- 14734005 TI - Testicular function in patients with testicular cancer treated with bleomycin etoposide-carboplatin (BEC(90)) combination chemotherapy. AB - OBJECTIVE: To investigate the impact of bleomycin-etoposide-carboplatin combination chemotherapy on long-term fertility in patients with testicular germ cell tumors. METHODS: Twenty-five patients with high risk stage I and IM non seminomatous germ cell tumors (NSGCT, Group A) and 44 with advanced seminoma or NSGCT (Group B) were treated with bleomycin 30 mg (days 2, 9, 16), etoposide 165 mg/m(2) (days 1-3) and carboplatin 400mg/m(2) or AUC 5 (day 1) (BEC(90)). Treatment was repeated every 3 weeks. Group A patients received 2 cycles of BEC(90), while Group B ones received 4 to 5 cycles of BEC(90). Sperm count and hormonal analyses were examined pre- and post-chemotherapy. Counts were classified as normospermia (NS) if >20 x 10(6)ml(-1), oligospermia (OS) if 1-20 x 10(6)ml(-1) and azoospermia (AS) if <1 x 10(6)ml(-1). RESULTS: Patients were followed for a median of 2.9 years post-chemotherapy. The post-orchidectomy median luteinizing hormone (LH) serum levels were slightly above the upper normal limit while the serum levels of follicle stimulating hormone (FSH) and testosterone (T) were within the reference interval. Thirty-eight (55%) patients had NS pre-chemotherapy. None of the 14 NS patients who received 2 cycles of BEC(90) had AS post-chemotherapy, while only 1 of the 24 NS patients who were treated with > or =4 cycles of BEC(90) had AS post-treatment. Among the NS patients, 93% and 83%, respectively, remained NS following chemotherapy. Overall, 90% of patients had recovery (61% NS, 29% OS) of spermatogenesis after treatment. The median FSH serum values were significantly elevated at least 1-year post chemotherapy when compared with the pre-treatment levels. Eighteen months post chemotherapy the median FSH values had returned to the reference limits. Serum LH and T levels were unaffected by treatment. The pre-treatment sperm count and the bulk of disease were significantly associated with recovery of spermatogenesis. No association was found between recovery of spermatogenesis and 2 or > or =4 cycles of chemotherapy, age > or =30 years and post-chemotherapy lymph node dissection. Thirteen patients (4 with OS) fathered 16 children. No congenital abnormalities occurred in any of these children. CONCLUSION: The BEC(90) regimen has no major effect on fertility and Leydig cell function. However, carboplatin based chemotherapy has been proved less effective than cisplatin-based chemotherapy and is not currently used in the treatment of testicular cancer. PMID- 14734006 TI - Liver harvesting surgical technique for the treatment of retro-hepatic caval thrombosis concomitant to renal cell carcinoma: perioperative and long-term results in 15 patients without mortality. AB - OBJECTIVE: Radical surgical treatment improves the prognosis of patients affected by Inferior Vena Cava (IVC) thrombosis concomitant to renal carcinoma. However, thrombus extension above the infrahepatic IVC represents a major technical topic for surgeons because of the possible occurrence of uncontrollable haemorrhages and tumor fragmentation. We report the results of an innovative surgical approach to caval thrombosis including the isolation of the IVC from the liver as routinely performed during liver harvesting. In the presence of retro-hepatic IVC thrombosis, this technique improves vascular control and allows to perform a large cavotomy with an en-bloc removal of the thrombus and the tumor. METHODS: From January 1995 through June 2003, 15 patients with renal cancer and caval thrombosis were treated at our Institution. Four, ten and one patients were respectively affected by an infrahepatic (Level I), retro-hepatic (Level II) and atrial (Level III) IVC thrombosis. RESULTS: All patients underwent radical surgical treatment. In presence of Level II caval thrombosis, the patients underwent the above reported surgical technique. Perioperative mortality was absent; major morbidity occurred in one patient (6.7%). The thrombus was radically removed in all cases. After a mean follow-up of 53.9 months (5-100 months) all patients but one are still alive. One patient died 9 months after surgery with multiple bilateral pulmonary metastases. CONCLUSIONS: Isolation of the retro-hepatic IVC is a safe and effective manoeuvre to significantly reduce perioperative mortality and morbidity in patients affected by Level II caval thrombosis concomitant to renal carcinoma. PMID- 14734007 TI - Transobturator tape (Uratape): a new minimally-invasive procedure to treat female urinary incontinence. AB - OBJECTIVE: Assessment of one-year results of a new technique of transobturator suburethral tape in the treatment of female stress urinary incontinence. METHODS: UraTape, a non-woven, non-elastic polypropylene tape with a 15 mm long central (suburethral) silicone-coated section was inserted via the transobturator route. The tape is inserted tension-free in a horizontal plane underneath the middle of the urethra between the two obturator foramens. The ends of the tape are tunnelled percutaneously with a tunneller. As the retropubic space is preserved intact, cystoscopy is not required. From May 2000 to February 2002, 150 patients with stress urinary incontinence without associated prolapse were operated and a minimum of 1 year follow-up was available for 32 patients (mean follow-up 17 months; range 13-29). The mean age was 64 years (range 50-81). All patients were assessed before surgery by clinical and urodynamic examination: 5 patients presented sphincter incompetence (maximum closure pressure <20 cm H(2)O); 5 patients presented with recurrent urinary incontinence after Burch procedure or TVT; 18 patients presented with mixed incontinence, six of them with detrusor instability confirmed by cystometry. The results were evaluated by two independent investigators (clinical examination, uroflowmetry, cough test). Voiding disorders suggesting bladder outflow obstruction were defined as the presence of the following two criteria: Q(max)<15 ml/s, residual urine volume >20%. RESULTS: 29/32 patients (90.6%) were cured and 3/32 (9.4%) were improved. Mean operating time was 15minutes. No intra-operative complications were recorded. One patient had complete postoperative bladder retention which resolved after 4 weeks of self-catheterization. There were no problems with urethral erosion, residual pain or functional impairment related to the tape. 5/32 patients had voiding disorders suggesting bladder outflow obstruction. Two patients developed de novo urge incontinence. CONCLUSION: Uratape transobturator tape is a simple and effective procedure with follow-up of one year for the treatment of female stress urinary incontinence confirmed after 1 year of follow up. The transobturator approach avoids the risk of bladder, bowel or vascular injuries. Evaluation of the results after a longer follow-up period is needed to validate this technique. PMID- 14734008 TI - Objective assessment of urinary incontinence in women: comparison of the one-hour and 24-hour pad tests. AB - OBJECTIVE: To compare the performance and clinical usefulness of the one-hour and 24-hour pad tests in terms of the relationship with reported symptoms and urodynamic diagnosis. METHODS: 341 women aged 40 years and over reporting lower urinary tract symptoms who were recruited to a nurse led continence service, and went on to receive urodynamics, a one-hour and a 24-hour pad test and completed a urinary diary. RESULTS: For both pad tests, there was a significant difference in the amount of urine leaked between the different urodynamic diagnoses (p<0.0001, for the one-hour and p=0.001 for the 24-hour test). Women with sphincter incompetence leaked significantly more than women with detrusor instability, or those with no abnormality. There was a significant difference between the proportion of women dry on a one-hour pad test and those dry on a 24-hour pad test (26.0% versus 38.4%, difference 12.4%; CI 5.5; 19.4). There is a positive relationship between amount of urine leakage and symptom severity expressed in terms of number of incontinent episodes for both pad tests. CONCLUSION: Both pad tests bore little relationship to the underlying urodynamic diagnosis but there was a positive relationship with symptom severity. The 24-hour pad test appears to be clinically a more useful too than the one-hour test. The two types of pad test are probably assessing incontinence in different ways. We suggest that the minimum data set should include structured questions, diaries and the 24-hour pad test. PMID- 14734009 TI - Does surgical repair of pelvic prolapse improve patients' quality of life? AB - OBJECTIVES: The aims of this study were to evaluate the efficacy of surgical repair in patients with pelvic prolapse, and to assess the postoperative quality of life (QOL). METHODS: A total of 70 patients (mean age: 66.7 years) underwent transvaginal two-corner bladder neck suspension in combination with transvaginal hysterectomy (63 cases), and anterior with (49) or without (21) posterior colporrhaphy. The status of recurrence and complications were followed in all patients at a mean follow-up period of 32.0 months. Postoperative patients' quality of life (QOL) consisting of four items (sensation of vaginal bulging, urinary incontinence, difficulty to urinate, and health-related QOL) was assessed in 52 cases whose were followed at least two years (mean: 41.6 months, range: 24.3-69.1). RESULTS: Sixty-eight patients (97%) were recurrence-free. Nine of the 39 (23%) patients with difficulty to urinate before surgery had persistent symptoms postoperatively. Multivariate analysis revealed that cystometric abnormalities, voiding symptoms at seventh days after surgery, and weak detrusor contraction were independent prognostic factors for persistent voiding symptoms. As for the QOL, all items had significant improvement at 13 months after surgery as compared to baseline condition. A longitudinal study showed improvement of these symptoms sustained at least up to four years. CONCLUSIONS: This study suggests that surgical repair can achieve results with long-term durability as well as improving the QOL. In addition, assessment of the detrusor function may be needed for patients who complain difficulty to urinate in avoiding persistent such symptoms. PMID- 14734010 TI - Treatment of anterior vaginal wall prolapse with porcine skin collagen implant by the transobturator route: preliminary results. AB - OBJECTIVE: To assess the feasibility and efficacy of porcine skin collagen (Pelvicol) implant by the transobturator route for the treatment of anterior vaginal wall prolapse. PATIENTS AND METHODS: Twenty-seven women with stage III or IV prolapse underwent anterior vaginal wall repair with porcine skin collagen implantation by the transobturator route, together with bilateral sacropinofixation; respectively 17 and 16 women underwent hysterectomy and bilateral sapingo-oophorectomy during the same procedure. The feasibility and peri/postoperative complications of the transobturator procedure were assessed, together with anatomical and functional outcome. RESULTS: Porcine skin collagen implantation was feasible in every case. The transobturator procedure lasted a median of 25 min (range 20-35). There were no vessel or bladder injuries and no postoperative complications (including infections). Median follow-up was 14 months (range 8 to 24). No rejection of the porcine grafts occurred. Twenty-two women (81%) had optimal anatomic results, while the remaining five women (19%) had persistent asymptomatic stage I or II anterior vaginal wall prolapse (1 and 4 cases, respectively). In one case the collagen implant had to be removed after one year because of persistent pain due to transfixing vaginal stitch. One patient with optimal anatomic results had recurrent stage III Ba prolapse at 18 months. Quality of life and urinary discomfort scores improved significantly after the procedure (p<0.0001 and p<0.005, respectively). CONCLUSION: These preliminary results suggest that porcine skin collagen implantation by the transobturator route is a safe and effective treatment for anterior vaginal wall prolapse. PMID- 14734011 TI - Laparoscopic surgery for pheochromocytoma. AB - OBJECTIVE: To evaluate the feasibility of laparoscopic adrenalectomy for pheochromocytoma. MATERIAL: Between January 1998 and March 2002, 12 patients aged from 19 to 76 (average age 54 years) underwent 13 adrenalectomies (7 right and 6 left, 1 bilateral) using laparoscopic surgery. A specific anti-hypertensive preparation was begun prior to the operation. Peaks of blood pressure were treated by bolus Nicardipine and sinus tachycardia by bolus Esmolol. Catecholamines were dosed at different times during the intervention. As far as surgery was concerned, the adrenalectomies were performed 11 times using the transperitoneal route and twice using the retroperitoneal route. The adrenal vein was found and ligatured before manipulation of the adrenal gland. RESULTS: Average length of operation was 127 minutes (75 to 195). Average blood loss was 105 ml (0 to 1000). Catecholamines dosed throughout showed a variable increase in plasma rates during peritoneal insufflation and manipulation of the gland. They were responsible for 5 cases of hypertensive bouts and 2 cases of tachycardia which were treated with medication. No surgical conversions were necessary. Average length of hospital stay was 4.18 days (3 to 6); average size of adrenal tumours was 44 mm (30 to 72); average follow-up, 18.4 months. CONCLUSION: This study showed the feasibility of adrenalectomy for pheochromocytoma using laparoscopic surgery, subject to specific medical preparation to reduce the consequences of peroperative bouts of hypertension and sinus tachycardia during peritoneal insufflation and manipulation of the adrenal gland despite initial ligature of the main adrenal vein. PMID- 14734012 TI - Microbial flora in ileal and colonic neobladders. AB - OBJECTIVE: To describe bacterial colonization in patients with ileal and colonic neobladders. METHODS: Twenty-three patients with right colon neobladders, 30 with ileal neobladders, 11 who had undergone radical prostatectomy, and 6 healthy controls were included. Culture of clean-catch, midstream urine specimens was done weekly for 3 weeks, and this was repeated after 6 months. Residual urine was measured, and the patients were interviewed about leakage. All patients and controls were antibiotic free during the study except for 13 of the ileal neobladder patients, who were treated with trimethoprim 100mg daily. RESULTS: Urine cultures from controls and prostatectomy patients were negative for bacteria, whereas 67% of the specimens from patients with neobladders, not on antibiotic therapy, were culture positive, and half of these contained uropathogenic species, such as Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, and Enterococcus faecalis. Bacterial colonization (including uropathogenic strains) was strongly correlated with residual urine (p<0.005), but not with leakage. Anaerobic strains were found more frequently (p=0.04) in urine from ileal neobladders than in urine from colonic neobladders. The 13 patients with ileal neobladders and on prophylactic antibiotic therapy carried bacteriuria in 80% of the samples, the majority being anaerobic strains. Uropathogenic strains, mainly Enterecoccus faecalis was revealed in 30% of the samples. CONCLUSIONS: The lower urinary tract of patients with ileal or colonic neobladders is heavily colonized with potentially uropathogenic and anaerobic bacteria. Complete bladder emptying reduces the bacterial burden. Anaerobic colonization is increased in neobladders reconstructed from ileum. Prophylactic antibiotic therapy does not seem to reduce the bacterial burden, but interferes with the bacterial composition. PMID- 14734013 TI - Retrospective analysis of efficacy and tolerability of tolterodine in children with overactive bladder. AB - OBJECTIVE: To evaluate the efficacy and tolerability of tolterodine in children with an overactive bladder, treated in a single incontinence centre. MATERIALS AND METHODS: A retrospective analysis of a database of a total of two hundred and fifty-six patients (175 boys and 81 girls, age range 3 years to 17 years, mean age 8.33 years) with urodynamically confirmed bladder overactivity was performed. All children received tolterodine tartrate (dose range of 0.5-4 mg orally). In group I (n=205) tolterodine tartrate replaced anticholinergic drugs (AC) (oxybutinin chloride or oxyphencyclimin hydrochloride). A subgroup of patients switched because of intolerance due to serious adverse events (60.4%) or because of lack of improvement in micturition variables (39.6%). In group II tolterodine was prescribed as initial therapy (n=51). Tolerability was assessed by a standardised questionnaire on adverse events at every outdoor clinic visit. Efficacy assessment was based on micturition diary variables, mean change of maximum bladder capacity and number of incontinence episodes/24 h. RESULTS: The mean treatment time was 9.32 months with a range from 1.5 months to 23.4 months. The final dose was 0.1mg/kg orally daily divided into two doses. In group I central nervous system disorders (81%) were the most common adverse events, 26.2% showed flushing, 12.2% accommodation problems and 25.2% had gastrointestinal complaints (constipation, encopresis, abdominal pain). Withdrawal of the non selective antimuscarinic drug resulted in total recovery from adverse events. Introduction of tolterodine in group I and II caused no serious adverse events. Nine patients (3.5%) reported side-effects and only two discontinued treatment. There were no reports of flushing, troubles of visual accommodation, hyperpyrexia. In group I we observed a mean decrease in urgency by 38.7%, a mean increase in maximal bladder capacity by 33.6% and the number of incontinence episodes decreased by 64.8%. In group II we observed equivalent values with a significant (p<0.001) change in maximal bladder capacity (49.7%), incontinence episodes (64.8%) and micturition episodes/24 h. CONCLUSIONS: The results of this retrospective analysis suggest that tolterodine is well tolerated in children and offers an effective treatment for urinary symptoms due to overactive bladder. Tolterodine is superior to non-selective antimuscarinic drugs, with respect to adverse events, allowing more compliance and more effective treatment in children. PMID- 14734014 TI - Androgen receptor regulation by physiological concentrations of the isoflavonoid genistein in androgen-dependent LNCaP cells is mediated by estrogen receptor beta. AB - OBJECTIVES: Isoflavonoids are discussed for use in chemoprevention and treatment of prostate cancer. We investigated the potential of genistein to modulate androgen receptor (AR) expression and transcriptional activity in the human androgen-sensitive prostate cancer cell line LNCaP. MATERIALS AND METHODS: AR expression at mRNA and protein level was analyzed by real-time RT-PCR and immunoblot, respectively. In conditioned media PSA was measured by a microparticle enzyme immunoassay (MEIA). Binding of genistein to the AR was tested in a radioligand-binding assay and reporter gene co-transfection assay was employed to investigate AR activity. RESULTS: Using concentrations of genistein that have been detected in sera of Asian men on regular soy-diet we found down regulation of androgen receptor at both mRNA and protein level. The relative binding affinity to the AR was below 4% when compared to methyltrienologe (R1881) and there was no modulation of AR transcriptional activity by genistein concentrations up to 1 microM. We also demonstrated inhibition of PSA secretion after genistein treatment. As the anti-estrogen ICI 164 384 abolished the inhibitory effect of genistein and ER-beta, but not ER-alpha is expressed in LNCaP cells we postulate that the mechanism of genistein action on androgen receptor is mediated through ER-beta. CONCLUSION: Using physiological concentrations of genistein we showed AR down-regulation by genistein in prostate cancer cells occurring via ER-beta. This likely results in a modified response to hormonal stimuli and may help to explain the low incidence of prostate cancer in the Asian population. PMID- 14734015 TI - The importance of a functional trehalose biosynthetic pathway for the life of yeasts and fungi. AB - The view of the role of trehalose in yeast has changed in the last few years. For a long time considered a reserve carbohydrate, it gained new importance when its function in the acquisition of thermotolerance was demonstrated. More recently the cellular processes in which the trehalose biosynthetic pathway has been implicated range from the control of glycolysis to sporulation and infectivity by certain fungal pathogens. There is now enough experimental evidence to conclude that trehalose 6-phosphate, an intermediate of trehalose biosynthesis, is an important metabolic regulator in such different organisms as yeasts or plants. Its inhibition of hexokinase plays a key role in the control of the glycolytic flux in Saccharomyces cerevisiae but other, likely important, sites of action are still unknown. We present examples of the phenotypes produced by mutations in the two steps of the trehalose biosynthetic pathway in different yeasts and fungi, and whenever possible examine the molecular explanations advanced to interpret them. PMID- 14734016 TI - Cyclic AMP signaling in Cryptococcus neoformans. AB - As pathogenic microorganisms establish an infection, they must be able to sense host-specific signals and respond by elaborating determinants that allow for survival in these hostile conditions. Pathogen cell surface proteins detect these signals and activate signal transduction cascades that ultimately alter gene expression resulting in an adaptive cellular response. Here we review the mechanisms by which a pathogenic fungus uses the highly conserved cAMP signal transduction pathway to regulate cellular differentiation as well as its virulence potential. PMID- 14734017 TI - Comparison of the epidemiology, drug resistance mechanisms, and virulence of Candida dubliniensis and Candida albicans. AB - Candida dubliniensis is a pathogenic yeast species that was first identified as a distinct taxon in 1995. Epidemiological studies have shown that C. dubliniensis is prevalent throughout the world and that it is primarily associated with oral carriage and oropharyngeal infections in human immunodeficiency virus (HIV) infected and acquired immune deficiency syndrome (AIDS) patients. However, unlike Candida albicans, C. dubliniensis is rarely found in the oral microflora of normal healthy individuals and is responsible for as few as 2% of cases of candidemia (compared to approximately 65% for C. albicans). The vast majority of C. dubliniensis isolates identified to date are susceptible to all of the commonly used antifungal agents, however, reduced susceptibility to azole drugs has been observed in clinical isolates and can be readily induced in vitro. The primary mechanism of fluconazole resistance in C. dubliniensis has been shown to be overexpression of the major facilitator efflux pump Mdr1p. It has also been observed that a large number of C. dubliniensis strains express a non-functional truncated form of Cdr1p, and it has been demonstrated that this protein does not play a significant role in fluconazole resistance in the majority of strains examined to date. Data from a limited number of infection models reflect findings from epidemiological studies and suggest that C. dubliniensis is less pathogenic than C. albicans. The reasons for the reduced virulence of C. dubliniensis are not clear as it has been shown that the two species express a similar range of virulence factors. However, although C. dubliniensis produces hyphae, it appears that the conditions and dynamics of induction may differ from those in C. albicans. In addition, C. dubliniensis is less tolerant of environmental stresses such as elevated temperature and NaCl and H(2)O(2) concentration, suggesting that C. albicans may have a competitive advantage when colonising and causing infection in the human body. It is our hypothesis that a genomic comparison between these two closely-related species will help to identify virulence factors responsible for the far greater virulence of C. albicans and possibly identify factors that are specifically implicated in either superficial or systemic candidal infections. PMID- 14734018 TI - The internal transcribed spacers and 5.8S rRNA gene show extensive diversity among isolates of the Cryptococcus neoformans species complex. AB - Sequences of the internal transcribed spacer (ITS) region including the 5.8S rRNA gene delineated seven genotypes within the three varieties of Cryptococcus neoformans via specific combinations of eight nucleotide differences located at positions 10, 11, 15, 19, 108 (ITS1), 221 (5.8S), 298 and 346 (ITS2). The ITS types correlated to polymerase chain reaction fingerprint/random amplification of polymorphic DNA (RAPD) molecular types: with ITS type 1 (ATACTAGC)=C. neoformans var. grubii, molecular types VNI+VNII and the serotype A allele of the AD hybrid, VNIIIA; ITS type 2 (ATATAGGC)=the serotype D allele of the AD hybrid, VNIIIB, and C. neoformans var. neoformans, VNIV; and ITS type 3 (GCGCTGGC) and ITS type 7 (ACGCTGGC)=VGI=RAPD type III, ITS type 4 (ACACTGAC)=VGII=RAPD type II, ITS type 5: (ACACTGGG)=VGIII=RAPD type I, ITS type 6 (ACACTGGC)=VGIV=RAPD type IV, all corresponding to C. neoformans var. gattii. Cloned sequences from serotype AD revealed that the hybrid serotype is diploid at the ITS1-5.8S-ITS2 locus carrying the ITS type 1 (ATACTAGC) and the ITS type 2 (ATATAGGC) alleles. ITS sequencing is a useful technique for genotyping the three C. neoformans varieties and for subtyping within C. neoformans var. gattii. PMID- 14734019 TI - Identification of a negative regulatory element which regulates basal transcription of a multidrug resistance gene CDR1 of Candida albicans. AB - We have earlier shown that transcriptional activation of the Candida drug resistance gene, CDR1, is linked to various stresses wherein a proximal promoter (-345 bp from the transcription start point (TSP)) was found to be predominantly more responsive. In this study we have examined basal expression of the CDR1 proximal promoter by employing a Renilla luciferase reporter system. We observed that upon sequential deletion of the proximal promoter, there was modulation in basal reporter activity. The reporter activity was highest (2.3-fold) in NGY261 ( 261 bp from TSP), and was reduced upon subsequent deletions. DNase I footprinting revealed four protected regions (W1, W2, W3 and W4) in the proximal promoter which could represent possible trans-acting factor binding sites and thus might be involved in CDR1 expression. Site-directed mutational analysis of three of these protected regions did not significantly affect the basal reporter activity, however, the mutation of W1 led to a considerable enhancement in reporter activity (approximately 4-fold) and was designated a negative regulatory element (NRE). Mutation as well as deletion of the W1 sequence in the native promoter ( 1147 bp from TSP) and sequential deletion of the 5'-flanking region-harboring W1 (NRE) also resulted in enhanced promoter reporter activity. When the reporter activity of native (NPY1147) and NRE-mutated (NGYM1147) promoter integrants was monitored throughout the growth phase of Candida albicans, there was modulation in reporter activity in both integrants, but interestingly the level of basal reporter activity of the NRE-mutated promoter was always approximately 3-fold higher than that of the native promoter. UV cross-linking and affinity purification confirmed that a purified approximately 55-kDa nuclear protein specifically interacts with the NRE. Taken together, we have identified a NRE and purified its interactive protein, which may be involved in controlling basal expression of CDR1. PMID- 14734020 TI - Expression analysis of the Candida albicans lipase gene family during experimental infections and in patient samples. AB - Secreted lipases of Candida albicans are encoded by a gene family with at least 10 members (LIP1-LIP10). The expression pattern of this multigene family was investigated using reverse transcription polymerase chain reaction in experimental infections and in samples of patients suffering from oral candidosis. The findings illustrate that individual lipase genes are differentially regulated in a mouse model of systemic candidosis with some members showing sustained expression and others being transiently expressed or even silent. The lipase gene expression profile depended on the stage of infection rather than on the organ localization. This temporal regulation of lipase gene expression was also detected in an experimental model of oral candidosis. Furthermore, the expression of candidal lipase genes in human specimens is shown for the first time. PMID- 14734021 TI - The histidine kinases of Candida albicans: regulation of cell wall mannan biosynthesis. AB - Previously, we have used both biochemical and immunological approaches to determine that the two-component, histidine kinase Chk1p regulates cell wall biosynthesis in Candida albicans. These data were obtained by comparing wild-type cells to a strain of C. albicans deleted in CHK1. The dysregulation of cell wall biosynthesis in the mutant reduces its adherence to human esophageal tissue and results in avirulence. In the current study, we used transmission immune electron microscopy (IEM) to visualize the cell surface of both wild-type (CAF2) and the chk1 mutant (CHK21). IEM was performed using two IgM monoclonal antibodies to either an acid-stable mannan epitope (Mab B6) or to an acid-labile mannan epitope (Mab B6.1). We observed that the cell surface of the CHK21 mutant was more reactive than wild-type cells with Mab B6, while the reactivity of Mab B6.1 was similar for both CAF2 and CHK21. These observations correlate with previous data on the Western blotting of mutant and wild-type cells using the same monoclonal antibodies, i.e., greater activity with Mab B6 than with Mab B6.1. In addition to CHK1, two other histidine kinases (SLN1 and NIK1) have been described in C. albicans. Mutants in both sln1Delta and nik1Delta were compared by Western blotting using Mab B6 and Mab B6.1. Reactivity of each mutant to Mab B6 was similar to that observed with the chk1 mutant; on the other hand, the mannoprotein profiles obtained with Mab B6.1 in all mutants were similar to wild type cells. We also compared the expression of 29 genes involved in mannan synthesis by reverse transcription-polymerase chain reaction (RT-PCR) and found that expression of a subset of six genes (ALG2, ALG6, ALG8, MNT3, PMT6, KRT2) was upregulated in all histidine kinase mutants, while increased expression of ALG7 was only observed in the sln1 and nik1 mutants, MNN1 was upregulated in the chk1 and nik1 mutants, and MNN4 was upregulated in the nik1Delta. Our data indicate that each of the C. albicans HK proteins may regulate similar functions in cell wall biosynthesis. This activity could be achieved in either a common or parallel, redundant signal transduction pathway(s). PMID- 14734022 TI - Yeast cells harboring human alpha-1,3-fucosyltransferase at the cell surface engineered using Pir, a cell wall-anchored protein. AB - Human alpha-1,3-fucosyltansferase (FucT) encoded by the FUT6 gene was displayed at the cell surface of yeast cells engineered using the yeast cell wall protein Pir1 or Pir2, and the FucT activity was detected at the surface of cells producing the Pir1-HA-FUT6 or Pir2-FLAG-FUT6 fusion proteins. To obtain higher activity, we engineered the host yeast cells in which endogenous PIR genes of the PIR1-4 gene family were disrupted. Among the disruptants, the pir1Delta pir2Delta pir3Delta strain with the PIR1-HA-FUT6 fusion gene showed the highest FucT activity, which was about three-fold higher than that of the wild-type strain. Furthermore, the co-expression of both the Pir1-HA-FUT6 and the Pir2-FLAG-FUT6 fusions showed an approximately 1.5-fold higher activity than that in the cell wall displaying Pir1-HA-FUT6 alone. The present method was thus effective for producing yeast cells that can easily synthesize various oligosaccharides, such as Le(x) and sLe(x), using Pir-glycosyltransferase fusions in combination with the deletion of endogenous PIR genes. PMID- 14734023 TI - Minisatellites in Saccharomyces cerevisiae genes encoding cell wall proteins: a new way towards wine strain characterisation. AB - With the aim of developing new tools for the characterisation of wine yeasts, by means of databases available on-line we scanned the genome of Saccharomyces cerevisiae in search of potentially polymorphic targets. As we have previously observed for SED1, we found that other genes coding for cell wall proteins contain minisatellite-like sequences. A polymerase chain reaction (PCR) survey of SED1 and three of these others, namely AGA1, DAN4 and HSP150, in a population of wild S. cerevisiae demonstrated that these genes are highly polymorphic in length and represent a sink of unexplored genetic variability. The primer pairs designed on the gene open reading frames yield stable and repeatable amplification profiles that show a level of resolution that allows the clear discriminate between different strains. These can therefore be utilised for PCR-based typing of S. cerevisiae. PMID- 14734024 TI - Identification of low-dye-binding (ldb) mutants of Saccharomyces cerevisiae. AB - We have completed the identification of Saccharomyces cerevisiae genes that are defective in previously isolated ldb (low-dye-binding) mutants. This was done by complementation of the mutant's phenotype with DNA fragments from a genomic library and by running standard tests of allelism with single-gene deletion mutants of similar phenotype. The results were as follows: LDB2 is allelic to ERD1; LDB4 to SPC72; LDB5 to RLR1; LDB6 to GON7/YJL184W; LDB7 to YBL006C; LDB9 to ELM1; LDB10 to CWH36; LDB11 to COG1; LDB12 to OCH1; LDB13 to VAN1; LDB14 to BUD32; and LDB15 to PHO85. Since the precise function of some of the genes is not known, these data may contribute to the functional characterization of the S. cerevisiae genome. PMID- 14734025 TI - Zinc finger transcriptional activators of yeasts. AB - Transcriptional transactivators are important proteins which in addition to controlling the cell regulatory circuitries, can be manipulated for various biotechnological processes. The latter is of great interest for non-conventional yeasts used for industrial purposes. To facilitate the identification of these transactivators, we have reanalyzed the "Genolevures" data (FEBS Lett. 487 (2000); http://cbi.labri.u-bordeaux.fr/Genolevures/) for the presence of zinc finger (Zf) proteins. After analysis of 239 RST ("random sequence tag") sequences, we describe in this paper 161 homologs of the Saccharomyces cerevisiae Zf proteins present in one or several of 13 different hemiascomyceteous yeasts. These partial sequences have been evaluated on different criteria such as percentage of identity of the proteins, synteny, detailed analysis of the Zf motif and flanking regions, and iterative BLASTs. They can be used to fetch the corresponding gene. PMID- 14734026 TI - ATG23, a novel gene required for maturation of proaminopeptidase I, but not for autophagy. AB - In rich media proaminopeptidase I is targeted to the vacuole via the Cvt pathway and during starvation via autophagy. We here identify Atg23 (Ylr431c), a protein of so far unknown function, as a novel component essential for proaminopeptidase I maturation under non-starvation conditions. Maturation of proaminopeptidase I takes place in starved atg23Delta cells. Selective vacuolar targeting of the autophagosomal marker GFP-Aut7 and the accumulation of autophagic bodies during starvation in the presence of phenylmethylsulfonyl fluoride suggest that autophagy occurs in atg23Delta cells but at a reduced rate. In atg23Delta cells mature vacuolar carboxypeptidase Y is present and accumulation of quinacrine suggests no significant defect in vacuolar acidification. Furthermore, growth of atg23Delta cells on nitrocellulose detects no significant secretion of carboxypeptidase Y. PMID- 14734027 TI - Cloning and characterization of the MAL11 gene encoding a high-affinity maltose transporter from Torulaspora delbrueckii. AB - The transport and regulation of maltose utilization by Torulaspora delbrueckii, one of the most abundant non-Saccharomyces species present in home-made corn and rye bread dough, has been investigated. A DNA fragment containing the MAL11 gene from T. delbrueckii (TdMAL11) was isolated by complementation cloning in Saccharomyces cerevisiae. DNA sequence analysis revealed the presence of an open reading frame (ORF) of 1884 bp, encoding a 627-amino acid membrane protein, which displays high homology to other yeast maltose transporters. Upstream of TdMAL11, the DNA insert contained a partial ORF (TdMAL12) on the opposite strand, which showed high similarity to the S. cerevisiae MAL12 gene. Sequence analysis, Northern blot and transport measurements indicated that TdMAL11 expression is regulated by the carbon source. Attempts to disrupt TdMAL11 revealed the presence of two functional MAL loci. Disruption of a single copy decreased the V(max) of maltose transport, but not the K(m), whereas the double disruption abolished the uptake of this sugar in T. delbrueckii. PMID- 14734028 TI - The SUV3 gene from Saccharomyces douglasii is a functional equivalent of its Saccharomyces cerevisiae orthologue and is essential for respiratory growth. AB - In the yeast Saccharomyces cerevisiae the product of the nuclear gene SUV3 has been shown to be involved in a variety of mitochondrial post-transcriptional processes. We have cloned and sequenced the SUV3 gene from Saccharomyces douglasii, a close relative of S. cerevisiae which has important changes in the organization of its mitochondrial genome and concomitant changes in nucleo mitochondrial interactions. We show that the S. douglasii SUV3 gene shares considerable structural homology (92% amino acid sequence identity) with its S. cerevisiae counterpart and that their nucleotide sequences display evidence of recent divergence. To determine the function of the S. douglasii SUV3 gene we have constructed a strain carrying an inactive SUV3 gene and analyzed the effect of this inactivation on the integrity of the mitochondrial genome and on the stability of mitochondrial transcripts. We have demonstrated that the S. douglasii SUV3 gene, like the S. cerevisiae gene, is essential for respiratory growth and for stability of the intron-containing mitochondrial transcripts, thus the two genes are functionally equivalent. Also the S. douglasii and S. cerevisiae SUV3 genes are completely interchangeable, despite the differences in the structure of the mitochondrial chromosome in the two yeasts. PMID- 14734029 TI - Ty1 insertions in intergenic regions of the genome of Saccharomyces cerevisiae transcribed by RNA polymerase III have no detectable selective effect. AB - The retrotransposon Ty1 of Saccharomyces cerevisiae inserts preferentially into intergenic regions in the vicinity of RNA polymerase III-transcribed genes. It has been suggested that this preference has evolved to minimize the deleterious effects of element transposition on the host genome, and thus to favor their evolutionary survival. This presupposes that such insertions have no selective effect. However, there has been no direct test of this hypothesis. Here we construct a series of strains containing single Ty1 insertions in the vicinity of tRNA genes, or in the rDNA cluster on chromosome XII, which are otherwise isogenic to strain 337, containing zero Ty1 elements. Competition experiments between 337 and the strains containing single Ty1 insertions revealed that in all cases, the Ty1 insertions have no selective effect in rich medium. These results are thus consistent with the hypothesis that the insertion site preference of Ty1 elements has evolved to minimize the deleterious effects of transposition on the host genome. PMID- 14734030 TI - The yeast Zygosaccharomyces bailii: a new host for heterologous protein production, secretion and for metabolic engineering applications. AB - Molecular tools for the production of heterologous proteins and metabolic engineering applications of the non-conventional yeast Zygosaccharomyces bailii were developed. The combination of Z. bailii's resistance to relatively high temperature, osmotic pressure and low pH values, with a high specific growth rate renders this yeast potentially interesting for exploitation for biotechnological purposes as well as for the understanding of the biological phenomena and mechanisms underlying the respective resistances. Looking forward to these potential applications, here we present the tools required for the production and the secretion of different heterologous proteins, and one example of a metabolic engineering application of this non-conventional yeast, employing the newly developed molecular tools. PMID- 14734031 TI - Heterologous expression of Zygosaccharomyces rouxii glycerol 3-phosphate dehydrogenase gene (ZrGPD1) and glycerol dehydrogenase gene (ZrGCY1) in Saccharomyces cerevisiae. AB - We examined the effects of heterologous expression of the open reading frames (ORF) of two genes on salt tolerance and glycerol production in a Saccharomyces cerevisiae strain deficient in glycerol synthesis (gpd1Deltagpd2Delta). When the ORF of the Zygosaccharomyces rouxii glycerol 3-phosphate dehydrogenase gene (ZrGPD1) was expressed under the control of the GAL10 promoter, salt tolerance and glycerol production increased; when the ORF of the glycerol dehydrogenase gene (ZrGCY1) was expressed under the control of the GAL1 promoter, no such changes were observed. Zrgcy1p had a weak effect on glycerol production. These results suggest that Zrgpd1p is the primary enzyme involved in Z. rouxii glycerol production, following a mechanism similar to that of S. cerevisiae (Gpd1p). When the ORFs of the S. cerevisiae glycerol 3-phosphatase gene (GPP2) and ZrGPD1 were simultaneously expressed, glycerol production increased, compared with that in yeast expressing only ZrGPD1. PMID- 14734032 TI - Stimulation of astaxanthin formation in the yeast Xanthophyllomyces dendrorhous by the fungus Epicoccum nigrum. AB - A fungal contaminant on an agar plate containing colonies of Xanthophyllomyces dendrorhous markedly increased carotenoid production by yeast colonies near to the fungal growth. Spent-culture filtrate from growth of the fungus in yeast-malt medium also stimulated carotenoid production by X. dendrorhous. Four X. dendrorhous strains including the wild-type UCD 67-385 (ATCC 24230), AF-1 (albino mutant, ATCC 96816), Yan-1 (beta-carotene mutant, ATCC 96815) and CAX (astaxanthin overproducer mutant) exposed to fungal concentrate extract enhanced astaxanthin up to approximately 40% per unit dry cell weight in the wild-type strain and in CAX. Interestingly, the fungal extract restored astaxanthin biosynthesis in non-astaxanthin-producing mutants previously isolated in our laboratory, including the albino and the beta-carotene mutant. The fungus was identified as Epicoccum nigrum by morphology of sporulating cultures, and the identity confirmed by genetic characterization including rDNA sequencing analysis of the large-subunit (LSU), the internal transcribed spacer, and the D1/D2 region of the LSU. These E. nigrum rDNA sequences were deposited in GenBank under accesssion numbers AF338443, AY093413 and AY093414. Systematic rDNA homology alignments were performed to identify fungi related to E. nigrum. Stimulation of carotenogenesis by E. nigrum and potentially other fungi could provide a novel method to enhance astaxanthin formation in industrial fermentations of X. dendrorhous and Phaffia rhodozyma. PMID- 14734033 TI - Sympatric natural Saccharomyces cerevisiae and S. paradoxus populations have different thermal growth profiles. AB - Saccharomyces cerevisiae and its close congener S. paradoxus are typically indistinguishable by the phenotypic criteria of classical yeast taxonomy, but they are evolutionarily distinct as indicated by hybrid spore inviability and genomic sequence divergence. Previous work has shown that these two species coexist in oak-associated microhabitats at natural woodland sites in North America. Here, we show that sympatric populations of S. cerevisiae and S. paradoxus from a single natural site are phenotypically differentiated in their growth rate responses to temperature. Our main finding is that the S. cerevisiae population exhibits a markedly higher growth rate at 37 degrees C than the S. paradoxus population; we also find possible differences in growth rate between these populations at two lower temperatures. We discuss the implications of our results for the coexistence of these yeasts in natural environments, and we suggest that thermal growth response may be an evolutionarily labile feature of these organisms that could be analyzed using genomic approaches. PMID- 14734034 TI - Correlations among measures of phenotypic and genetic variation within an oligotrophic asexual yeast, Candida sonorensis, collected from Opuntia. AB - Many descriptions of yeast species are based on a limited number of strains collected at one time from a single locale. Often, little is known of phenotypic or genotypic variation and covariation within species. We compare 36 strains of an asexual cactophilic yeast, Candida sonorensis, collected from Opuntia cacti. Comparisons were based on geographical distances between collection locales, responses to physiological assimilation and stress tests, random amplified polymorphic DNA (RAPD) profiles, partial Lage Subunit (LSU) rDNA sequences, relative DNA-DNA hybridization values, and electrokaryotypes. There was significant variation among strains in all types of data collected. Comparisons among the different data types found significant positive associations between RAPD profiles, geographical distances, physiologies, reassociation values, and electrokaryotypes. No significant associations were found between rDNA sequences and any other type of variation measured. Based on RAPD, reassociation, electrokaryotype, and physiological data, the 36 strains could be divided into two groups: those collected in West Texas (nine strains) and all others. RAPD data indicated that 10 (of 12) Australian strains also formed a distinct clade. The taxonomic and phylogenetic status of these clades is discussed. Evidence that new genotypes can sweep through large geographic areas is also discussed. PMID- 14734036 TI - Reinstatement of Rhodotorula colostri (Castelli) Lodder and Rhodotorula crocea Shifrine & Phaff, former synonyms of Rhodotorula aurantiaca (Saito) Lodder. AB - Rhodotorula aurantiaca (Saito) Lodder is an anamorphic basidiomycetous yeast species that belongs to the so-called "Erythrobasidium lineage" of the Urediniomycetes, according to molecular phylogenetic studies based on nucleotide sequence analyses of different ribosomal DNA regions. In the most recent editions of the yeast taxonomy treatises the species Rhodotorula colostri (Castelli) Lodder and Rhodotorula crocea Shifrine & Phaff were listed as synonyms of R. aurantiaca. Taxonomic heterogeneity within R. aurantiaca was demonstrated in a study based on whole-cell protein profiles and is also hinted at by the observed differences in physiological and biochemical characteristics among the different strains under that species name. We determined partial nucleotide sequences of the 26S rRNA gene (D1/D2 domains) of strains maintained in the CBS culture collection under R. aurantiaca, including the type strains of its synonyms. The results showed that R. colostri and R. crocea are clearly distinct from R. aurantiaca and from any other currently recognised basidiomycetous yeast species. Furthermore, phylogenetic analysis of the sequence data placed the former two species in separate lineages of the Microbotryomycetidae: R. colostri in the "ruineniae clade" (Sporidiobolus lineage or Sporidiobolales) and R. crocea loosely linked to Rhodotorula javanica (Microbotryum lineage). PMID- 14734035 TI - Characterisation and classification of phylloplane yeasts from Portugal related to the genus Taphrina and description of five novel Lalaria species. AB - Taphrina Fries is a genus of dimorphic ascomycetes comprising more than 90 species distinguished by the specific infections they produce on different vascular plants. Their filamentous states are restricted to parasitised plant tissue whereas the yeast states are saprobic and can be grown on artificial media. The latter coincide with the anamorphic phases and have been given separate nomenclatural status by the erection of the genus Lalaria R.T. Moore. In its original circumscription Lalaria included only 23 yeast states of known species of Taphrina and its creation was then redundant. Here we describe five novel species in the genus Lalaria to accommodate a total of 44 yeast isolates obtained mainly from leaf surfaces (phylloplane) of different plants in Portugal: Lalaria arrabidae sp. nov. (one strain), L. carpini sp. nov. (one strain), L. inositophila sp. nov. (37 strains), L. kurtzmanii sp. nov. (one strain) and L. veronaerambellii sp. nov. (four strains). L. inositophila was notable for its widespread occurrence since it was recovered during two consecutive years from the leaves of miscellaneous plant species. In the absence of sexual states and of unequivocal associations to particular host plants, the taxonomic relationship of the novel species to the yeast states of Taphrina available from culture collections was verified by the comparative analysis of physiological and molecular characteristics. The latter included PCR fingerprinting using single primers for microsatellite regions, sequencing of the 5' end of the 26S rRNA (LSU) gene (D1/D2 domains) and of the ITS1 and ITS2 rDNA spacer regions, and DNA DNA hybridisation experiments. An emended description of the genus Lalaria is provided. PMID- 14734037 TI - The XVIth Meeting on the Biology of Kluyveromyces, Cortona, Italy, 12-14 September 2003. PMID- 14734038 TI - The 4th Brewing Yeast Fermentation Performance Congress, Oxford, England, 9-12 September, 2003. PMID- 14734039 TI - Failing human myocardium "fails" to translate performance demand signals. PMID- 14734040 TI - Na+/Ca2+ exchange linking beta2-adrenergic G(i) signaling to heart failure: associated defect of adrenergic contractile support. PMID- 14734041 TI - Thrombin: beyond coagulation. PMID- 14734042 TI - Unbalanced RANKL/RANK pathway in aortic valve sclerosis. PMID- 14734043 TI - The acid test. PMID- 14734044 TI - Re-evaluating therapeutic neovascularization. AB - Numerous animal studies have established that neo-vascularization of ischemic tissue can be enhanced with exogenous growth factors and small clinical studies have shown encouraging results. However, the two largest randomized clinical trials to date were negative. Mechanistically, the major stimuli for neo vascularization are hypoxia and inflammation. Hypoxia-inducible-factor (HIF-1) is a 'master switch' protein that is generated in response to hypoxia and binds to more than 40 hypoxia sensitive genes, inducing a panoply of angiogenic and protective metabolic responses. Inflammatory signals recruit T-lymphocytes and macrophages into areas of neo-vascularization which act as a source of angiogenic and arteriogenic factors. Although hypoxia and inflammation are interdependent in eliciting neo-vascular responses, angiogenesis appears to be hypoxia-dependent, whereas inflammation and hemodynamic factors drive arteriogenesis. The negative outcome of the two largest trials may have many reasons. There are issues relating to patient selection, choice of growth factor therapy, dosing and route administration, concomitant medication, trial design including the efficacy parameters that were selected and a lack of sufficient insight into the mechanisms that are responsible for neo-vascularization. In order to move forward the therapeutic objective should be switched to arteriogenesis although this process is even more poorly understood than angiogenesis. Genetic studies in mice with intrinsically different arteriogenic responses combined with studies in human populations with differences in the extent of collateral development may provide fundamental insight into arteriogenic mechanisms. Attention should also be focused on the way in which arteriogenesis is stimulated and the endpoints of clinical trials should be redefined. PMID- 14734045 TI - Abnormal frequency-dependent responses represent the pathophysiologic signature of contractile failure in human myocardium. AB - BACKGROUND: - The normal increase in isometric developed force (DF) with faster pacing rates, known as the positive force-frequency response/relationship (FFR), is altered in failing myocardium, as shown by its negative response to increased pacing. The objective of this study was to determine if increasing Ca2+ influx with L-type Ca2+ channel (L-CaCh) agonists: BayK 8644 (BayK) and FPL 64176 (FPL) or increased extracellular Ca2+ could increase contractility and normalize the FFR in failing myocardium. METHODS: - Isometric DF was measured in right ventricular trabeculae from failing (n = 28) and non-failing (n = 12) human hearts at various stimulation frequencies (0.5-2.5 Hz) before and after bath application of BayK (250 nM), FPL (100 nM), or high Ca2+ (7.0 mM). Post-rest (PR) experiments were also conducted on several trabeculae. RESULTS: - In trabeculae from failing hearts, the DF decreased with an increase in pacing. Addition of L CaCh agonists increased DF to similar levels in trabeculae from both failing and non-failing hearts at slow pacing rates, but did not alter the negative FFR in the failing group. During increased rest intervals, the amount of PR potentiation was diminished in trabeculae from failing hearts as compared to the non-failing preparations. CONCLUSION: - This study demonstrates that the abnormal FFR observed in trabeculae from failing hearts is a reliable physiologic signature of the cardiomyopathic state even when DF, at slow stimulation frequencies, is relatively high. These studies further demonstrate that the impaired FFR is not due to an inability to further increase contractility. Rather, our findings suggest that the abnormal FFR and blunted PR potentiation alike are a reflection of an altered functional balance between Ca2+ re-uptake and Ca2+ extrusion. PMID- 14734046 TI - Loss of beta-adrenoceptor response in myocytes overexpressing the Na+/Ca(2+) exchanger. AB - Increased Na+/Ca(2+)-exchanger (NCX) and altered beta-adrenoceptor (betaAR) responses are observed in failing human heart. To determine the possible interaction between these changes, we investigated the effect of NCX overexpression on responses to isoproterenol in adult rat ventricular myocytes. Responses to isoproterenol were largely mediated through the beta1AR in control myocytes. Adenovirally-mediated overexpression of NCX, at levels, which did not alter basal contraction of myocytes, markedly depressed the isoproterenol concentration-response curve. Responses to isoproterenol could be restored to normal by beta2AR blockade, suggesting a beta2AR-mediated inhibition of beta1AR signalling. Pertussis toxin normalised isoproterenol responses in NCX cells, indicating that beta2AR effects were mediated by Gi. Negative-inotropic effects of high concentrations of ICI 118,551, previously shown to be due to beta2AR-Gi coupling, were increased in NCX cells. We conclude that NCX upregulation can markedly alter the consequences of betaAR stimulation and that this may contribute to the alterations in betaAR response seen in failing human heart. PMID- 14734047 TI - Reactive oxygen species-sensitive p38 MAPK controls thrombin-induced migration of vascular smooth muscle cells. AB - Thrombin has been implicated in the development of atherosclerosis and restenosis, in which migration of vascular smooth muscle cells (VSMC) is a crucial event. Thrombin-stimulated VSMC migration is associated with increased generation of reactive oxygen species (ROS), activation of mitogen-activated protein kinases (MAPKs), and production of growth factors and chemoattractants. In this study, we examined the interrelation of these signals to determine the pathway controlling thrombin-directed migration of human VSMC. Our results show that thrombin stimulated the production of ROS and activation of p38 MAPK. ROS were required for thrombin-induced VSMC migration since both generation of ROS and cell migration were significantly attenuated by inhibitors of NAD(P)H oxidase, diphenyleneiodonium (DPI) and apocynin (Apo.), and by the hydrogen peroxide scavenger, catalase (Cat.). Activation of p38 MAPK by thrombin was inhibited by DPI, Apo. and Cat., indicating ROS are used as messengers for activating this kinase. p38 MAPK is an important step since SB 203580, a selective inhibitor of p38 MAPK, suppressed the cell migration induced by thrombin. Furthermore, thrombin increased the expression of vascular endothelial growth factor (VEGF), a chemoattractant for VSMC, and this expression was inhibited by DPI, Apo., Cat. and SB 203580. Addition of anti-VEGF antibody significantly attenuated thrombin-induced migration. Collectively, the data presented here show that thrombin has stimulated VSMC migration and VEGF expression through an ROS-sensitive p38 MAPK pathway. VEGF synthesized and released by the cell served as a secondary mediator in thrombin-directed migration. PMID- 14734048 TI - Receptor activator of nuclear factor kappaB ligand and osteoprotegerin regulate aortic valve calcification. AB - OBJECTIVE: - Recent studies have suggested that valvular calcification in calcific aortic stenosis (AS) may be actively regulated. "Receptor Activator of Nuclear factor kappaB Ligand" (RANKL) and osteoprotegerin (OPG) are members of a cytokine system involved in bone turnover and vascular calcification. Their role in calcific AS is not known. METHODS AND RESULTS: - By immunohistochemistry using human aortic valves, RANKL was not expressed at relevant levels in controls but detectable in AS. OPG expression was marked in controls but significantly lower in AS. Areas containing focal calcification exhibited significantly less OPG positive cells as compared to non-calcified regions. Stimulation with RANKL lead to a significant rise in matrix calcification, nodule formation, alkaline phosphatase activity, expression of the bone-type isoenzyme of alkaline phosphatase, and expression of osteocalcin in cultured human aortic valve myofibroblasts. Moreover, RANKL increased DNA binding of the essential osteoblast transcription factor cbfa-1. CONCLUSION: - RANKL and OPG are differentially expressed in calcific AS. In cultured human aortic valve myofibroblasts, RANKL promotes matrix calcification and induces the expression of osteoblast-associated genes, indicating a transition towards an osteogenic phenotype. These results suggest that the RANKL-OPG pathway may regulate valvular calcification in calcific AS? PMID- 14734049 TI - Phospholamban is required for CaMKII-dependent recovery of Ca transients and SR Ca reuptake during acidosis in cardiac myocytes. AB - Initially during acidosis, Ca transient amplitude (Delta[Ca]i) and the rate constant of [Ca]i decline (k(Ca)) are decreased, but later during acidosis Delta[Ca]i and k(Ca) partially recover. This recovery in rat myocytes could be inhibited by KN-93 suggesting that CaMKII-dependent protein phosphorylation (and enhanced SR Ca uptake) may be responsible. To test whether phospholamban (PLB) is required for the Delta[Ca]i and k(Ca) recovery during acidosis, we used isolated myocytes from PLB knockout (PLB-KO) vs. wild-type (WT) mice. [Ca]i was measured using fluo-3. During the initial phase of acidosis (1-4 min), Delta[Ca]i decreased in WT myocytes (n = 8) from 1.75 +/- 0.19 to 1.10 +/- 0.13 DeltaF/F0 (P < 0.05) and k(Ca) decreased from 3.20 +/- 0.22 to 2.38 +/- 0.18 s(-1) (P < 0.05). Later during acidosis (6-12 min), Delta[Ca]i partially recovered to 1.41 +/- 0.18 DeltaF/F0 and k(Ca) to 2.78 +/- 0.22 s(-1) (i.e. both recovered by approximately 50%). CaMKII inhibition using KN-93 completely prevented this recovery of Delta[Ca]i and k(Ca) during late acidosis in WT myocytes. In PLB-KO myocytes (n = 11) Delta[Ca]i decreased during early acidosis from 2.92 +/- 0.31 to 1.33 +/- 0.17 DeltaF/F0 (P < 0.05) and k(Ca) decreased from 10.45 +/- 0.56 to 7.58 +/- 0.68 s(-1) (P < 0.05). However, Delta[Ca]i did not recover during late acidosis and k(Ca) decreased even more (6.59 +/- 0.65 s(-1)). Parallel results were seen for contractile parameters. We conclude that PLB is crucial to the recovery of Delta[Ca]i and k(Ca) during acidosis. Moreover, PLB phosphorylation by CaMKII plays an important role in limiting the decline in Ca transients (and contraction) during acidosis. PMID- 14734050 TI - Intracellular angiotensin II fusion protein alters AT1 receptor fusion protein distribution and activates CREB. AB - In recently published studies, we show that angiotensin II (AII) generated from an engineered rat angiotensinogen cDNA, and maintained intracellularly, is growth stimulatory for a rat hepatoma cell line. In the present study, we report that co expression of AII fused to cyan fluorescent protein (ECFP/AII) and angiotensin type I receptor fused to yellow fluorescent protein (AT1R/EYFP) enhances proliferation of COS-7 and CHO-K1 cells by 59% and 64%, respectively, compared to cells expressing the corresponding independent proteins (P < 0.001 for both). This effect is inhibited by losartan, suggesting (as in our previous published studies) that losartan is internalized by the cells, via receptor-mediated endocytosis, and thus inhibits intracellular receptor-ligand interaction. The growth effect is independent of anti-AII antibodies suggesting that it does not reflect AII secretion into the culture media; AII is also undetectable in the media. Expression of AT1R/EYFP with ECFP/AIIC (control scrambled sequence AII fused to ECFP) has no effect upon cell proliferation. ECFP/AII also alters the cellular localization of AT1R/EYFP. ECFP/AII is concentrated in the nucleus, but shows diffuse cytoplasmic fluorescence as well. AT1R/EYFP, expressed independently, is visible in the endoplasmic reticulum and Golgi apparatus of COS 7 and CHO-K1 cells as early as 24-h post-transfection. At 72 h, it is visibly associated with the plasma membrane. By 144 h, 85% of the cells show detectable circumferential fluorescence. In contrast, in cells that express AT1R/EYFP and ECFP/AII, both proteins accumulate in the nucleus and only 13% of the cells show visible plasma membrane-associated yellow fluorescence at 144 h (P < 0.001). Furthermore, co-expression of ECFP/AII with AT1R/EYFP stimulates cAMP response element-binding protein (CREB) activity in CHO-K1 and COS-7 cells. Exogenous AII similarly significantly increases CREB activation in AT1R/EYFP-stably transfected CHO-K1 and COS-7 cells. PMID- 14734051 TI - Inducibility of life-threatening ventricular arrhythmias is related to maximum left ventricular thickness and clinical markers of sudden cardiac death in patients with hypertrophic cardiomyopathy attributable to the Asp175Asn mutation in the alpha-tropomyosin gene. AB - We investigated inducibility of life-threatening arrhythmias with programmed ventricular stimulation (PVS) in relation to clinical markers of sudden cardiac death (SCD) in subjects with hypertrophic cardiomyopathy (HCM) attributable to the Asp175Asn mutation in the alpha-tropomyosin gene (TPM1-Asp175Asn). PVS was performed with up to three extrastimuli and distribution of markers of SCD was evaluated in 21 adult subjects with the TPM1-Asp175Asn. Sustained polymorphic ventricular tachycardia (VT) or ventricular fibrillation (VF) was induced in seven of 21 subjects (33%). Inducible subjects had more severe left ventricular hypertrophy (LVH) and an increased number of markers of SCD (family history of SCD, syncope or presyncope, fall in systolic blood pressure (BP) during exercise, documented non-sustained VT (NSVT), and marked LVH) compared to non-inducible subjects (IVS 2.4 +/- 0.3 cm vs. 1.6 +/- 0.5 cm, P < 0.001; and two to three vs. one to two markers of SCD, P = 0.007, respectively). In conclusion, in HCM attributable to the Asp175Asn mutation in the alpha-tropomyosin gene, life threatening arrhythmias were induced in one third of the patients. Inducibility was associated with the maximum left ventricular (LV) thickness and the number of markers of SCD, suggesting that in HCM patients with an identical causative mutation, susceptibility to ventricular arrhythmias is related to the cardiomyopathic phenotype. PMID- 14734052 TI - Functional role of connexin43 gap junction channels in adult mouse heart assessed by inducible gene deletion. AB - The gap junction protein Connexin43 (Cx43) is expressed in various cell types during embryonic development and in adult mice. Cx43 null mice (Cx43-/-) die perinatally due to cardiac malformation. In order to define the major functional role of Cx43 gap junction channels in adult mice and to circumvent perinatal death as well as direct or indirect compensation of Cx43 deficiency during development, we established a novel conditional Cx43 mouse mutant. To ablate Cx43 in adult mice in all cells that express Cx43 at a certain time, we targeted the 4 hydroxytamoxifen inducible Cre recombinase, Cre-ER(T), into the endogenous Cx43 locus. This approach left only one Cx43 coding region to be deleted upon induction of Cre-ER(T) activity. Highly efficient inducible ablation of Cx43 was shown in an embryonic stem cell test system and in adult mice. Although Cx43 protein was decreased in different tissues after induction of Cre-ER(T)-mediated recombination, cardiac abnormalities most likely account for death of those mice. Surface and telemetric ECG recordings revealed significant delay of ventricular activation and death during periods of bradyarrhythmia preceded by tachycardias. This novel approach of inducible ablation of Cx43 highlights the functional importance of normal activation of ventricular cardiomyocytes mediated by Cx43 gap junction channels in adult mouse heart to prevent initiation of fatal arrhythmias. The new mouse model should be useful for further analyses of molecular changes initiated by acute loss of Cx43 expression in various cell types. PMID- 14734053 TI - Functional expression of L- and T-type Ca2+ channels in murine HL-1 cells. AB - In the search for a readily available source of native cardiac cells, we investigated the molecular and pharmacological properties of the immortalized cardiac atrial myocyte cell line, HL-1 cells. This work focused on the expression pattern of voltage-gated Ca2+ channels (VGCC). Reverse transcription-polymerase chain reaction analysis revealed that HL-1 cells have mRNA for several types of Ca2+ channels including the L-types, alpha1C and alpha1D, as well as T-types, alpha1H and alpha1G, but are lacking N-type, alpha1B and the T-type, alpha1I. Western blot analysis demonstrated significant alpha1C protein subunit expression, with less alpha1D subunit apparent, while alpha1A, alpha1B and alpha1E subunit expression was undetectable. Immunocytochemical staining showed that the alpha1C protein subunit is expressed predominantly on the cell surface, whereas the alpha1D protein is expressed mostly intracellularly. Whole-cell patch clamp measurements demonstrated the presence of low (ICa,T) and high (ICa,L) voltage-activated Ca2+ currents, with preferential sensitivity to mibefradil and nimodipine, respectively. Addition of increasing external Ca2+ concentrations, [Ca2+]o, resulted in Ca2+ influx measured by fluorometric imaging with an EC50 of 0.8 mM [Ca2+]o. At a fixed [Ca2+]o of 0.125 mM, Ca2+ influx was also triggered by increasing the extracellular K+ concentration, [K+]o, with an EC50 of 3.7 mM [K+]o. As increasing [K+]o depolarizes the cell, this latter result is consistent with Ca2+ influx through a voltage-dependent mechanism. L-type (nimodipine and verapamil) and T-type (mibefradil and pimozide) Ca2+ channel blockers inhibited Ca2+ influx with IC50s of 1, 2, 0.4 and 0.2 microM, respectively. Antagonists of N-type (omega-conotoxins GVIA) and P/Q-type (MVIIC or omega-agatoxin IVA) did not inhibit Ca2+ influx, consistent with the lack of expression of N-, P-, or Q-type channels observed in the molecular studies. Taken together, these findings indicate that HL-1 cells express L- and T-subtypes of VGCC and are a unique in vitro model system for the study of native, mammalian cardiac Ca2+ channels. PMID- 14734054 TI - Cardiomyocyte-specific desmin rescue of desmin null cardiomyopathy excludes vascular involvement. AB - Mice deficient in desmin, the muscle-specific member of the intermediate filament gene family, display defects in all muscle types and particularly in the myocardium. Desmin null hearts develop cardiomyocyte hypertrophy and dilated cardiomyopathy (DCM) characterized by extensive myocyte cell death, calcific fibrosis and multiple ultrastructural defects. Several lines of evidence suggest impaired vascular function in desmin null animals. To determine whether altered capillary function or an intrinsic cardiomyocyte defect is responsible for desmin null DCM, transgenic mice were generated to rescue desmin expression specifically to cardiomyocytes. Desmin rescue mice display a wild-type cardiac phenotype with no fibrosis or calcification in the myocardium and normalization of coronary flow. Cardiomyocyte ultrastructure is also restored to normal. Markers of hypertrophy upregulated in desmin null hearts return to wild-type levels in desmin rescue mice. Working hearts were perfused to assess coronary flow and cardiac power. Restoration of a wild-type cardiac phenotype in a desmin null background by expression of desmin specifically within cardiomyocyte indicates that defects in the desmin null heart are due to an intrinsic cardiomyocytes defect rather than compromised coronary circulation. PMID- 14734055 TI - Adenovirus-mediated overexpression of catalase attenuates oxLDL-induced apoptosis in human aortic endothelial cells via AP-1 and C-Jun N-terminal kinase/extracellular signal-regulated kinase mitogen-activated protein kinase pathways. AB - In a variety of vascular disorders, endothelial cells (ECs) are exposed to high levels of reactive oxygen species (ROS) generated intercellularly. Recently, several anti-oxidants, including catalase, have been suggested to be cytoprotective against the development of atherosclerosis. The object of this study was to investigate whether adenovirus-mediated gene transfer of catalase in ECs can attenuate ROS production and cell apoptosis under oxidized low density lipoprotein (oxLDL) stimulation. Adenovirus-mediated gene transfer of human catalase gene (Ad-Cat) resulted in a high level of catalase overexpression in human arterial EC (HAEC), which manifested a time-dependent increase in cell viability under the exposure of oxLDL and decreased oxLDL-induced apoptosis. Phosphorylation studies of ERK1/2, JNK, and p38, three subgroups of mitogen activator protein kinase demonstrated that catalase overexpression suppressed JNK phosphorylation and increased ERK1/2 phosphorylation. NF-kappaB and AP-1 were induced after the exposure of HAECs to oxLDL. While catalase overexpression was found to inactivate AP-1, it had no effect on NF-kappaB activity. These results provide the evidence that overexpression of catalase in ECs attenuates ROS production and cell apoptosis under oxLDL stimulation. The protective effect is mediated through the downregulation of JNK and the upregulation of ERK1/2 phosphorylation as well as AP-1 inactivation. This observation supports the feasibility of catalase gene transfer to human endothelium to protect against oxidant injury. PMID- 14734056 TI - Metabolic dysfunction and depletion of mitochondria in hearts of septic rats. AB - Our previous studies indicate that hearts from septic rats have decreased work with oxygen wasting. The present studies test if there is energy deficit, changes in cardiac mitochondrial content and caspase activation during sepsis. Anesthetized, male Sprague-Dawley rats received no surgical treatment (control), laparotomy (sham), or laparotomy with cecal ligation and puncture (CLP) to induce polymicrobial septic shock. Hearts were isolated 12-14 h later. Cardiac work, oxygen consumption, substrate oxidation and energy stores were measured in perfused hearts. Normalized density of mitochondria was determined in ventricles without perfusion by morphometric analysis with electron microscopy. Citrate synthase activity was assessed in homogenates and isolated mitochondria. Cardiac work decreased significantly in CLP (47%), while oxygen consumption and glucose oxidation were unchanged compared with control or sham hearts (oxygen and substrate wasting). Tissue adenosine triphosphate, creatine phosphate and glycogen were lower in CLP hearts (energy deficit). Mitochondrial grid intersects decreased significantly from 151 +/- 8 sham to 130 +/- 4 CLP out of 361 possible intersects and autophagy was observed in CLP hearts. Total activity of citrate synthase decreased in homogenates (99 +/- 8 micromol/min/g wet weight sham vs. 62 +/- 7 CLP, P < 0.05) and in the mitochondrial fraction (27 +/- 1 micromol/min/g wet weight sham to 22 +/- 1 CLP, P < 0.05). Calculated mitochondrial content decreased from 63 +/- 4 mg protein/g wet weight sham to 46 +/- 5 CLP, P < 0.05 (mitochondrial depletion). Caspase-3 activity doubled and tumor necrosis factor alpha content tripled in CLP hearts. CONCLUSIONS. - Oxygen and substrate wasting in CLP occurs with fewer mitochondria and energy deficit, processes that are coincident with caspase-3 activation. PMID- 14734057 TI - Mesoridazine: an open-channel blocker of human ether-a-go-go-related gene K+ channel. AB - Mesoridazine, a phenothiazine antipsychotic agent, prolongs the QT interval of the cardiac electrocardiogram and is associated with Torsade de pointes-type arrhythmias. In this study, we examined the effects of mesoridazine on human ether-a-go-go-related gene (HERG) K+ currents. HERG channels were stably expressed in human embryonic kidney 293 cells and studied using standard whole cell patch-clamp technique (37 degrees C). Mesoridazine blocked HERG currents in a concentration-dependent manner (IC50 550 nM at 0 mV); block increased significantly over the voltage range where HERG activates and saturated at voltages eliciting maximal HERG channel activation. Tonic block of HERG current by mesoridazine (1.8 microM) was minimal (< 2-4%). The rate of the onset of HERG channel block was rapid and dose dependent (tau = 54 +/- 7 ms at 0 mV and 1.8 microM mesoridazine), but not significantly affected by test potentials ranging from -30 to +30 mV. The V1/2 for steady-state activation was shifted from -31.2 +/- 1.0 to -39.2 +/- 0.5 mV (P < 0.01). The apparent rate of HERG channel deactivation was significantly reduced (fast tau = 153 +/- 8 vs. 102 +/- 6 ms at 50 mV, P < 0.01; slow tau = 1113 +/- 63 vs. 508 +/- 27 ms, P < 0.01). The inactivation kinetics and voltage dependence of steady-state inactivation of the HERG channel were not significantly altered by mesoridazine. These findings demonstrate that mesoridazine is a potent and rapid open-channel blocker of HERG channels. This block would explain the QT prolongation seen clinically at therapeutic concentrations (0.3-3.6 microM). PMID- 14734058 TI - Role of connexin 43 in ischemic preconditioning does not involve intercellular communication through gap junctions. AB - Connexin 43 (Cx 43) has recently been implicated in protection of ischemic preconditioning. Cx 43 colocalization with protein kinase C and p38 mitogen activated protein kinase is increased in preconditioned myocardium, Cx 43 phosphorylation is preserved in preconditioned myocardium, and hearts from Cx 43 deficient mice cannot be preconditioned. It is, however, unclear whether the important role of Cx 43 relates to intercellular communication through gap junctions or its function in volume homeostasis. To address this issue, we used isolated cardiomyocytes, which no longer-form gap junctions, from wild-type (n = 5) and heterozygous Cx 43-deficient mice (n = 8) and subjected them to 2 h simulated ischemia (hypoxia, acidosis) and an additional challenge by extracellular hypo-osmolarity (from 310 to 250 mOsm/l). Viability (trypan blue exclusion) was well maintained in normoxic wild-type cardiomyocytes (54 +/- 5% at baseline vs. 46 +/- 4 (mean +/- S.D.) % at 2 h). With simulated ischemia, viability was reduced to 17 +/- 5%. Preconditioning by a preceding exposure to 10 min simulated ischemia and 15 min reoxygenation preserved viability after 2 h simulated ischemia (36 +/- 1%, P < 0.001 vs. simulated ischemia). In Cx 43 deficient cardiomyocytes, viability was also well maintained in normoxia (56 +/- 10% vs. 44 +/- 10%). Viability was also reduced to 17 +/- 6% with 2 h simulated ischemia. In contrast to wild-type cells, preconditioning did not prevent the reduction in viability (18 +/- 8%). In conclusion, Cx 43 is essential for preconditioning in the absence of gap junctions, supporting its function through improved volume regulation. PMID- 14734059 TI - Antenna procedure for the correction of hook nail deformity. AB - A total of seven antenna procedures for hooknail deformity were performed between 1990 and 2000. At a mean follow-up of 3 years 4 months there were no major complications, all patients had an improvement in their deformity (subjectively and objectively) and were pleased with their results. Some residual deformity remained in all cases including shortening of the nail, contour defect of the pulp and flattening or residual curvature of the nail. PMID- 14734060 TI - Pulmonary and biceps function after intercostal and phrenic nerve transfer for brachial plexus injuries. AB - This pseudo-randomized study was performed to compare the pulmonary function and biceps recovery after intercostal (19 cases) and phrenic (17 cases) nerve transfer to the musculocutaneous nerve for brachial plexus injury patients with nerve root avulsions. Pulmonary function was assessed pre-operatively and postoperatively by measuring the forced vital capacity, forced expiratory volume in 1 second, vital capacity, and tidal volume. Motor recovery of biceps was serially recorded. Our results revealed that pulmonary function in the phrenic nerve transfer group was still significantly reduced 1 year after surgery. In the intercostal nerve transfer group, pulmonary function was normal after 3 months. Motor recovery of biceps in the intercostal nerve group was significantly earlier than that in phrenic nerve group. We conclude that pulmonary and biceps functions are better after intercostal nerve transfer than after phrenic nerve transfer in the short term at least. PMID- 14734061 TI - The role of manual occupation in the aetiology of Dupuytren's disease in men in England and Wales. AB - We compared the incidence of significant Dupuytren's disease in men across occupational social classes in England and Wales, using data from the National Morbidity Survey. We found that manual occupational social class was not associated with an increased incidence of Dupuytren's disease. In fact, the incidence rates of Dupuytren's disease in the elderly were higher in non-manual than in manual social classes. PMID- 14734062 TI - Proximal interphalangeal joint release in Dupuytren's disease of the little finger. AB - We present a prospective study, with 3-year follow-up, of the role and outcome of fasciectomy plus sequential surgical release of structures of the proximal interphalangeal joint in Dupuytren's contracture of the little finger. Our treatment programme involves fasciectomy for all patients followed by sequential release of the accessory collateral ligament and volar plate as necessary. Of the 19 fingers in the study, eight achieved a full correction by fasciectomy alone, and in these cases there was a fixed flexion deformity of 6 degrees at 3 months and 8 degrees at 3 years. The remaining 11 fingers (initial mean deformity 70 degrees flexion) were left with a fixed flexion deformity of 42 degrees after fasciectomy which reduced to 7 degrees with capsulo-ligamentous release. This increased to 26 degrees at 3 months but then remained relatively stable, increasing only to 29 degrees at 3 years. In our experience sequential proximal interphalangeal joint release has led to consistently good results with few complications in the correction of severe Dupuytren's disease of the little finger. PMID- 14734063 TI - A prospective randomised clinical trial of the intra-operative use of 5 fluorouracil on the outcome of dupuytren's disease. AB - 5-Fluorouracil reduces proliferation rates of fibroblasts, myofibroblast differentiation and contractility of ocular fibroblasts in vitro. This double blind randomized clinical trial assesses whether intra-operative topical treatment with 5-fluorouracil reduces the recurrence rate after limited excision of Dupuytren's tissue. Patients with two-digit disease were randomized to having 5-fluorouracil (25mg/ml) treatment for 5 minutes on one digit and placebo on the other. Fifteen patients were enrolled with 18 months follow-up. There were no peri-operative complications. Wound healing was not delayed and there was no deterioration in the flexion deformity of the 5-fluorouracil treated digits. Patients were subsequently assessed by joint angle measurement at 3, 6, 12 and 18 months. There was no significant difference between control and 5-fluorouracil treated digits. PMID- 14734064 TI - The use of Acufex wedge tag tissue anchors in hand surgery. AB - Over a period of 4 years, in various circumstances commonly seen in hand surgery, 100 patients underwent 127 soft tissue attachments to bone using the Acufex wedge tag system (Acufex Microsurgical, Inc, Mansfield, MA), a non-metallic bone anchor. No failures to maintain the attachment of the desired soft tissue to bone were identified. While less robust than the Mitek anchor, the other commonly available system of bone anchoring, and therefore possibly inappropriate for general orthopaedics, the Acufex wedge tag proved adequate for the smaller forces of hand surgery. PMID- 14734065 TI - Prevalence and epidemiological variation of anomalous muscles at guyon's canal. AB - Ultrasound was performed in 116 wrists of asymptomatic volunteers to determine the prevalence and morphology of anomalous muscles in Guyon's canal in the normal population. The size of the muscle was correlated with sex, hand-dominance and occupation. Anomalous muscles were identified in 47% of volunteers and 35% of wrists (male prevalence (50%); female prevalence (21%)). All were variants of abductor digiti minimi. Bilateral muscles were present in 50% of index cases in both sexes. Mean muscle thickness was 1.7 mm overall with no inter-sex variation. Muscle thickness did not vary with hand dominance or manual employment. The prevalence of anomalous muscles is higher than previously reported and may be sex linked. The size of the muscle may be an important factor in determining whether an anomalous muscle is significant in cases of ulnar nerve compression at Guyon's canal. PMID- 14734066 TI - Randomized trial of buffered versus plain lidocaine for local anaesthesia in open carpal tunnel decompression. AB - We performed a randomized double-blind case-control study in 64 consecutive patients undergoing open carpal tunnel decompression under local anaesthetic to assess the pain experienced on injection of plain lidocaine (pH 6.4) compared with lidocaine buffered with sodium bicarbonate (pH 7.4). The results showed no statistical difference in the pain scores reported by patients. The mean pain scores for all patients were low, and most patients reported that they were "not at all anxious" about receiving a similar injection in the future. PMID- 14734067 TI - Carpal ligament decompression under local anaesthesia: the effect of lidocaine warming and alkalinisation on infiltration pain. AB - This study investigated the effects of alkalinization and warming of lidocaine 1% on injection pain in patients undergoing carpal tunnel decompression. Sixty-four adult patients were randomly allocated into one of three groups: Group A (n=20) received plain lidocaine 1%, Group B (n=22) alkalinized lidocaine and Group C (n=22) warmed and alkalinized lidocaine. Pain on needle insertion and on infiltration was assessed using a 100mm Visual Analogue Scale (VAS). There was no significant difference regarding pain on needle insertion whereas significant differences were noted in reference to infiltration pain. In Groups B and C (alkalinized lidocaine) the VAS scores on skin infiltration were significantly lower than in Group A, while the pain score in Group C (alkalinized and warmed lidocaine) was significantly lower than in Group B. PMID- 14734068 TI - Relief operation in the case of paralysis of the median nerve. 1921. PMID- 14734069 TI - Transplantation of the M. abductor dig. V. where there is no ability to oppose the thumb. PMID- 14734070 TI - Changes in key pinch strength after excision of the trapezium and total joint arthroplasty. AB - Key pinch force was measured preoperatively and at follow-up (mean 25 months) in patients treated for basal joint arthritis by either trapziectomy with ligament reconstruction and tendon interposition (n=26) or total joint arthroplasty (n=27). There was a significant increase in key pinch strength with both treatments, but no difference between the two treatments. Total joint arthroplasty does not appear to produce stronger key pinch than trapziectomy with ligament reconstruction and tendon interposition. PMID- 14734071 TI - The osteology of the trapezium. AB - The present anatomical and clinical literature is not detailed enough for a clear understanding of the three-dimensional anatomy of the trapezium. It lacks descriptions of identifiable landmarks needed for the interpretation of two dimensional radiographs. Fifty dry cadaver trapezia were assessed and an extended surface anatomy described. New consistent landmarks were described and the tubercle of the trapezium was redefined. The incidence of the salient osteological features in Caucasian trapezia was recorded. PMID- 14734072 TI - MR imaging of thumb carpometacarpal joint ligament injuries. AB - This study describes the MR imaging appearances of the supporting ligaments of the thumb carpometacarpal joint in asymptomatic volunteers and in a group of patients following joint injury. Fourteen patients with 11 acute and three chronic injuries underwent MR imaging. The anterior oblique ligament was the most commonly injured ligament, usually on the metacarpal side where it was disrupted, or allowed dislocation because of subperiosted stripping from the base of the thumb metacarpal. The dorsal radial ligament was occasionally avulsed or partially torn from the trapezoid. Following chronic injury, MR imaging can evaluate ligamentous laxity, ganglion cyst formation or osteoarthritis. Accurate evaluation of ligament injury may identify patients who would benefit from surgery. PMID- 14734073 TI - Three-dimensional video analysis of forearm rotation before and after combined pronator teres rerouting and flexor carpi ulnaris tendon transfer surgery in patients with cerebral palsy. AB - The effect of combined pronator teres rerouting and flexor carpi ulnaris transfer on forearm rotation was prospectively studied by comparison of pre- and postoperative three-dimensional analysis of forearm range of motion in ten patients with cerebral palsy. One year postoperatively, surgery had improved maximal supination of the forearm in all patients by an average of 63 degrees, but there was also a mean loss of 40 degrees pronation. Forearm range of motion increased by a mean of 23 degrees. The centre of the range of motion on average shifted 52 degrees in the direction of supination. Based on these results of objective forearm range of motion analysis, we conclude that the common combination of pronator teres rerouting and flexor carpi ulnaris transfer in patients with cerebral palsy effectively facilitates active supination but impairs active pronation. PMID- 14734074 TI - A model for the conservative management of mallet finger. AB - This prospective study assessed the results of a custom-made thermoplastic splint for treatment of mallet finger deformity. From April 1999 to April 2000, 42 patients with mallet finger deformity were recruited. All patients were seen within 1 week and treated with a thermoplastic splint custom-made by the hand therapy department. The splint was simple to make, easy to fit and suitable for all finger shapes and sizes. It improved the deformity in 30 out of 34 cases, and caused no skin irritation. PMID- 14734075 TI - Solitary enchondroma at the hand. Long-term follow-up study after operative treatment. AB - Twenty-one patients (17 women and four men) who underwent operative treatment for a solitary enchondroma of the hand were examined at a follow-up of between 2 and 18 years (mean, 9 years). Radiographs showed normal cancellous bone at the site of surgery in 11 cases, three had recurrent enchondroma and seven had bone defects so that recurrence could not be excluded. Two of the three recurrences underwent reoperation. Previous studies have regarded persistent bony defects as evidence of complete excision without recurrence. However, in view of the slow asymptomatic growth of this tumour this opinion is incorrect. As shown in this study, recurrences may occur in these defects many years after excision surgery and go undetected until they cause widening or cortical erosion. We recommend periodical radiological re-examination for asymptomatic recurrences before weakness of bone leads to pathological fracture. PMID- 14734076 TI - Giant cell tumours of the tendon sheath: lack of correlation between nm23-H1 expression and recurrence. AB - Some authors attribute recurrences of giant cell tumours to biological factors which are only expressed in some tumours. Grover et al. (1998) suggested that the risk for recurrence is associated with the down-regulation of the nm23-H1 gene. We reviewed the charts of the 154 patients operated on for giant cell tumours of the tendon sheath and selected a group of patients with recurrence (ten cases) and a group of patients who did not have a recurrence after a minimum follow-up of 3 years (13 cases). Immunohistochemical detection of nm23-H1 was performed blindly of the clinical outcome on the paraffin-embedded specimens of these patients and no correlation was found between nm23-H1 expression and the risk for recurrence. PMID- 14734077 TI - Distraction lengthening of thumb metacarpal. AB - The purpose of this retrospective study was to evaluate the clinical outcome of distraction lengthening of the thumb metacarpal without bone grafting in seven patients with traumatic thumb loss. The distraction was stopped after 57 (range, 42 to 91) days, giving a median lengthening of 28 (range, 20-36)mm. It took an average of 155 (range, 118-196) days for bony consolidation to occur. The mean pinch power was 72% of that of the uninjured hand. The two-point discrimination on the pulp of the reconstructed thumb was 10 (range, 8-12)mm. There were no major complications. PMID- 14734078 TI - Dynamic external fixation for complex intraarticular phalangeal fractures. AB - Intraarticular phalangeal fractures are difficult to treat. The advantages of using dynamic external fixation devices include distraction of impacted fracture fragments and reduction in joint stiffness by allowing early joint mobilization. Previous reports have concentrated on pilon fractures and dorsal fracture dislocations affecting the proximal interphalangeal joint. We report our experience using a dynamic external spring fixator in the management of 15 patients with a variety of fracture patterns affecting the metacarpophalangeal, proximal interphalangeal and distal interphalangeal joints. In three cases the device spanned two adjacent joints. Long-term follow-up has shown excellent range of joint movement and no major complications. We consider that this device is an effective tool in the management of a range on complex intraarticular phalangeal fractures. PMID- 14734079 TI - Average versus maximum grip strength: which is more consistent? AB - The average of three consecutive measurements is the most frequently used method for grip strength assessment. The purpose of this study was to compare the consistency of the maximum value with that of the average value of three consecutive measurements of grip strength. One hundred healthy volunteers participated in this study. Three measurements of grip strength were taken on two occasions separated by 2 weeks. For each hand, two average values and two maximum values were obtained. Ninety-five per cent limits of agreement for the average method were - 8.3 (-23%) to +7.2 (+20%)kg and for the maximum method were - 8.8 ( 23%) to + 8 (+21%)kg. Both methods of grip strength assessment were found to be highly consistent with no statistically significant difference. PMID- 14734080 TI - Nomenclature in the Journal of Hand Surgery. AB - Unambiguous description of anatomical structures is essential to communicate information either in the clinical or research context. In view of this and following recommendations by the Nomenclature Committee of the International Societies for Surgery of the Hand (IFSSH), the booklet "Terminology for Hand Surgery" was published in 2001. The aim of this study was to identify whether the nomenclature used in the papers published in the Journal of Hand Surgery (British and European volume) followed its recommendations. The study showed that we still continue to use traditional terminology. The most common "mistake" is numbering of the metacarpals instead of naming them. Usage of newer terms would improve the clarity of information distributed. PMID- 14734081 TI - A molten metal, high-pressure injection injury of the hand. AB - A case of high-temperature, high-pressure injection injury of the hand is reported. The injury is particularly severe in that it involves both a high temperature thermal insult and a pressure effect, either of which, alone, can result in major disability. It is evident that the high temperature of the injected material considerably reduced the pressure required for penetration of the tissues. PMID- 14734082 TI - Palmar dislocation of the metacarpophalangeal joint of the finger. AB - We report a case of palmar dislocation of a finger metacarpophalangeal joint. Disruption of all the supporting structures of this joint and rupture of the flexor tendon sheath caused marked instability. Treatment was by open reduction and repair of the collateral ligaments. PMID- 14734083 TI - Re: Palmer plate fixation of AO type C2 fracture of distal radius using a locking compression plate. PMID- 14734085 TI - The biology of hematopoietic stem cells. AB - Rarely has so much interest from the lay public, government, biotechnology industry, and special interest groups been focused on the biology and clinical applications of a single type of human cell as is today on stem cells, the founder cells that sustain many, if not all, tissues and organs in the body. Granting organizations have increasingly targeted stem cells as high priority for funding, and it appears clear that the evolving field of tissue engineering and regenerative medicine will require as its underpinning a thorough understanding of the molecular regulation of stem cell proliferation, differentiation, self renewal, and aging. Despite evidence suggesting that embryonic stem (ES) cells might represent a more potent regenerative reservoir than stem cells collected from adult tissues, ethical considerations have redirected attention upon primitive cells residing in the bone marrow, blood, brain, liver, muscle, and skin, from where they can be harvested with relative sociological impunity. Among these, it is arguably the stem and progenitor cells of the mammalian hematopoietic system that we know most about today, and their intense study in rodents and humans over the past 50 years has culminated in the identification of phenotypic and molecular genetic markers of lineage commitment and the development of functional assays that facilitate their quantitation and prospective isolation. This review focuses exclusively on the biology of hematopoietic stem cells (HSCs) and their immediate progeny. Nevertheless, many of the concepts established from their study can be considered fundamental tenets of an evolving stem cell paradigm applicable to many regenerating cellular systems. PMID- 14734086 TI - Identification and isolation of hematopoietic stem cells. AB - Hematopoietic stem cells (HSCs) are defined by their ability to repopulate all of the hematopoietic lineages in vivo and sustain the production of these cells for the life span of the individual. In the absence of reliable direct markers for HSCs, their identification and enumeration depends on functional long-term, multilineage, in vivo repopulation assays. The extremely low frequency of HSCs in any tissue and the absence of a specific HSC phenotype have made their purification and characterization a highly challenging goal. HSCs and primitive hematopoietic cells can be distinguished from mature blood cells by their lack of lineage-specific markers and presence of certain other cell-surface antigens, such as CD133 (for human cells) and c-kit and Sca-1 (for murine cells). Functional analyses of purified subpopulations of primitive hematopoietic cells have led to the development of several procedures for isolating cell populations that are highly enriched in cells with in vivo stem cell activity. Simplified methods for obtaining these cells at high yield have been important to the practical exploitation of such advances. This article reviews recent progress in identifying human and mouse HSCs and current techniques for their purification. PMID- 14734087 TI - Biology of human hematopoietic stem and progenitor cells present in circulation. AB - Circulating hematopoietic stem and progenitor cells play important roles in the physiology and homeostasis of the hematopoietic system. The frequency of these cells varies throughout development, being more abundant during gestation. In the adult, the numbers of such cells are extremely low; however, they can be increased by intravenous administration of chemotherapy and/or recombinant cytokines to individuals. This mechanism--known as mobilization--involves the disruption of the interactions between primitive hematopoietic cells and microenvironment elements (stromal cells and extracellular matrix molecules), which are mediated by a group of molecules known as cell adhesion molecules. During the last two decades, circulating cells of newborns (those present in umbilical cord blood) and adults (mobilized peripheral blood) have gained relevance not only because of their biology, but also because of their clinical application. Indeed, at present the number of mobilized peripheral blood-derived hematopoietic cell transplants performed worldwide is clearly superior to the number of bone marrow transplants being done annually. On the other hand, the number of cord blood transplants has significantly increased during the last few years, and cord blood banking has expanded in a significant manner over the last decade. Circulating stem and progenitor cells are being manipulated ex vivo, both in cellular and molecular terms, and the clinical use of these manipulated cells is just beginning. Undoubtedly, hematopoietic cells present in circulation will play a key role in the development of both gene and cellular therapies for a variety of diseases. PMID- 14734088 TI - Telomeres, telomerase, and hematopoietic stem cell biology. AB - Telomeres are composed of the tandem DNA repeats and associated proteins that cap the end of linear chromosomes. They provide stability to the chromosome and protect against DNA loss associated with cellular replication. Telomeres are maintained by the reverse transcriptase, telomerase. The regulation of telomere length and telomerase activity is a complex and dynamic process that is tightly linked to cell cycle regulation. Hematopoietic stem cells have an impressive but finite proliferative potential and demonstrate telomeric shortening during replicative aging despite expression of low levels of telomerase. Recently, the important role of telomeres in human illness has been highlighted by studies of the rare genetic disorder dyskeratosis congenita. Here we review the role of telomeres and telomerase in the function and regulation of the hematopoietic stem cell compartment and their importance in hematologic disease. PMID- 14734089 TI - Hematopoietic stem cells in chronic myeloid leukemia. AB - Chronic myeloid leukemia (CML) is a malignant myeloproliferative disorder originating from a pluripotent hematopoietic stem cell that acquires a Philadelphia (Ph) chromosome encoding the BCR-ABL oncogenic fusion protein. This molecular abnormality that is thought to be causative in CML was the first acquired chromosome translocation associated with a human malignancy. This chromosomal translocation also makes it possible to precisely distinguish between residual normal (i.e., Ph-, BCR-ABL-) progenitor or stem cells and their leukemic counterpart, Ph+ or BCR-ABL+ progenitor/stem cells in every given sample of a patient with CML. This has provided seminal insights into the molecular and cellular biology of leukemia and also of the process of normal hematopoiesis. CML has become a fascinating model disease for malignancy in general. PMID- 14734090 TI - Human acute myeloid leukemia stem cells. AB - Acute myeloid leukemia (AML) is a clonal disorder defined by the accumulation of abnormally differentiated myeloid cells that are not mature; any myeloid lineage can be affected and the extent of maturation of the leukemia blasts can also vary. Because mature blast cells of AML have very limited proliferative capacity, it is believed that the leukemic clone is perpetuated by a rare population of leukemia stem cells (LSC) that have acquired a dramatic increase in their ability to self-renew. Elucidating the nature of the target cell that undergoes leukemic transformation and the resultant LSC that can initiate and maintain AML is essential for both the understanding of the leukemogenic process and for the design of effective therapies. However, identifying such cells using only clinical data from human subjects has been difficult due to obvious restriction in experimental intervention in humans. In addition, before clinical symptoms are presented, it is virtually impossible to acquire a complete picture of the early events in leukemogenesis. Other experimental approaches involved the study of naturally occurring or induced animal (murine) leukemias. While many aspects of these animal leukemias reproduced the human disease, there were also inconsistencies. The advent of xenotransplantation to accurately model human AML growing within an animal system has provided an important tool to begin to answer the fundamental questions regarding AML. This review will examine the work done using the xenograft system to characterize the nature of the leukemic clone and will specifically highlight the advances made in phenotypically, molecularly, and functionally defining the LSC. Finally, a variety of novel AML therapeutics aimed at eradicating the LSC will be discussed. PMID- 14734091 TI - Lingering biologic dilemmas about the status of the progenitor cells in myelodysplasia. AB - Myelodysplastic syndromes (MDS) are considered to be stem cell disorders. High incidence of intramedullary apoptosis has been associated with the peripheral cytopenia and refractory anemia in these disorders. The investigations on the cell of origin in the bone marrow have invariably been hampered by a poor yield of CD34+ cells from these marrows. Interestingly, even though limited in number, these studies raised more questions and dilemmas than providing answers. While the enigma surrounding the clonality of these marrows continues, the controversies regarding incidence of apoptosis, proliferation, and potential of clonogenic expansion may be closer to a settlement. The present review proposes a model depicting interplay between extraneous apoptogenic factors and intracellular apoptosis-susceptibility determinants that contributes significantly toward the progression of MDS and how a shift in dynamics of this interplay may provide grounds to accumulate additional mutations with a probable block in differentiation eventually leading to a leukemic transformation. PMID- 14734092 TI - Hematopoietic stem cells in aplastic anemia. AB - Profound cytopenia involving all blood lineages, a hallmark of aplastic anemia (AA), can result in devastating morbidity and high mortality. Although various etiologies and distinct pathophysiologic mechanisms may be involved, a profound defect in the stem cell compartment is a unifying feature in most patients with AA. As a stem cell disease, AA is very instructive and provides insights into the function and quantity of normal hematopoietic stem cells and their ability to regenerate. Pathophysiologically, understanding of AA may reveal mechanisms as to the evolution of other related bone marrow failure syndromes such as paroxysmal nocturnal hemoglobinuria and myelodysplasia-clonal diseases of hematopoiesis associated with defective stem cells. Conversely, constitutional forms of AA occurring in association with Fanconi anemia and dyskeratosis congenita demonstrate the role of specific genes and pathways in the dysfunction of the stem cells leading to the failure of the stem cell compartment. The acquired mechanisms resulting in depletion of stem cells in AA may involve fundamental pathways such as apoptosis and senescence as well as exhaustion of proliferative capacity or excessive differentiation. Inherent in the paucity of the bone marrow in AA, the study of the stem cells in AA has been very difficult due to their natural rarity and disease-specific contraction of the stem cell pool. Despite these scientific challenges, laboratory studies and systematic clinical observation provide valuable information of significance beyond its specific application to AA. PMID- 14734093 TI - Hematopoietic cell transplantation: five decades of progress. AB - During the past 50 years, the role of allogeneic hematopoietic cell transplantation (HCT) has changed from a desperate therapeutic maneuver plagued by apparently insurmountable complications to a curative treatment modality for thousands of patients with hematologic diseases. Now, cure rates following human leukocyte antigen (HLA) allogeneic HCT with matched siblings exceed 85% for some otherwise lethal diseases, such as chronic myeloid leukemia, aplastic anemia, or thalassemia. In addition, the recent development of non-myeloablative conditioning and stem cell transplantation has opened the way to include elderly patients with a wide variety of hematologic malignancies. Further progress in adoptive transfer of T cell populations with relative tumor specificity would make the transplant procedure more effective and would extend the use of allogeneic HCT for treatment of non-hematopoietic malignancies. PMID- 14734094 TI - Hematopoietic stem cell transplantation: bone marrow vs. mobilized peripheral blood. AB - Peripheral blood stem cells have largely replaced bone marrow as the source of cells in autologous transplantation because of more rapid neutrophil and platelet recovery and faster immune reconstitution. Allogeneic peripheral blood stem cells similarly lead to faster hematologic recovery: however, their effects on graft vs.-host disease, relapse, survival, and immune reconstitution have been less certain. Eight randomized trials have been published to date comparing the clinical outcomes of allogeneic-related donor bone marrow transplantation (BMT) vs. PBSCT and will be reviewed. In addition, comparisons between the two stem cell sources in unrelated donor transplantation and the increasingly utilized nonmyeloablative transplantation will be discussed. PMID- 14734095 TI - Non-myeloablative bone marrow transplantation. AB - As a result of the evolution of knowledge in the area of allogeneic hematopoietic stem cells (HSC) transplantation, several dogmata have been broken. We now have the following information: a) successful engraftment if allogeneic HSC bone marrow ablation of the recipient is not required; b) HSC create their own space through graft-vs.-host reactions; c) several malignancies are eradicated by the graft-vs.-tumor effect; d) allografting can be conducted on an out-patient basis; e) allografting can be done in aged or debilitated individuals; f) allografting can be achieved without transfusion of blood products, and g) costs of the allografting procedures can be substantially diminished. Breaking all these dogmata has resulted in availability of HSC allografting to a larger number of individuals, thus offering true curative therapeutic options to patients who otherwise would not qualify to receive these opportunities. PMID- 14734096 TI - Human embryonic stem cells: multilineage differentiation and mechanisms of self renewal. AB - Human ES (hES) cells are pluripotent stem cells isolated from the inner cell mass (ICM) of blastocysts, with the theoretical capacity to differentiate in vitro to produce all somatic and germ cell types. The diverse differentiation repertoire of hES cells makes them ideal candidates for the generation of tissues for transplantation therapies and drug discovery. However, to realize the full potential of hES cells it will be necessary to characterize the mechanisms that control self-renewal and differentiation into specific cell types. We review here the recent developments to differentiate human ES cell into lineages including neural and cardiac. Further, by reference to the self-renewal system established in murine ES we will discuss the possible mechanisms of self-renewal in hES cells. PMID- 14734097 TI - Mesenchymal stem cells. AB - It has become clear that adult mammalian bone marrow contains not one but two ostensibly discrete populations of adult stem cells. The first and by far the most fully characterized are the hematopoietic stem cells responsible for maintaining lifelong production of blood cells. The biological characteristics and properties of the second marrow resident population of stem cells, variously termed bone marrow stromal cells or mesenchymal stem cells, are in contrast much less well understood. In vitro, cultures established from single-cell suspensions of bone marrow from a wide range of mammalian species generate colonies of adherent marrow stromal cells, each derived from a single precursor cell termed a colony-forming unit-fibroblast (CFU-F). Culture conditions have been developed to expand marrow stromal cells in vitro while maintaining the capacity of these cells to differentiate into bone, fat, and cartilage. A significant portion of our current knowledge of this population of cells is based on analysis of the properties of these culture expanded cells, not on the primary colony-initiating cells. In this article, we will focus on methodologies to prospectively isolate stromal progenitors from mouse and human bone marrow and will review current data that suggest stromal progenitors in the bone marrow in situ are associated with the outer surfaces of blood vessels and may share identity with vascular pericytes. PMID- 14734098 TI - Neural stem cells in development and regenerative medicine. AB - In the last 10 years, enormous interest in neural stem cells has arisen from both basic and medical points of view. The discovery of neurogenesis in the adult brain has opened our imagination to consider novel strategies for the treatment of neurodegenerative diseases. Characterization of neurogenesis during development plays a fundamental role for the rational design of therapeutic procedures. In the present review, we describe recent progress in the characterization of embryo and adult neural stem cells (NSCs). We emphasize studies directed to determine the in vivo and in vitro differentiation potential of different NSC populations and the influence of the surrounding environment on NSC-specific differentiation. From a different perspective, the fact that NSCs and progenitors continuously proliferate and differentiate in some areas of the adult brain force us to ask how this process can be affected in neurodegenerative diseases. We propose that both abnormal cell death activation and decreased natural neuronal regeneration can contribute to the neuronal loss associated with aging, and perhaps even with that occurring in some neurodegenerative diseases. Furthermore, although NSC activation can be useful to treat neurodegenerative diseases, uncontrolled NSC proliferation, survival, and/or differentiation could cause tumorigenesis in the brain. NSC-mediated therapeutic procedures must take into account this latter possibility. PMID- 14734099 TI - Genetic modification of hematopoietic stem cells: recent advances in the gene therapy of inherited diseases. AB - Hematopoietic stem cells constitute a rare population of precursor cells with remarkable properties for being used as targets in gene therapy protocols. The last years have been particularly productive both in the fields of gene therapy and stem cell biology. Results from ongoing clinical trials have shown the first unquestionable clinical benefits of immunodeficient patients transplanted with genetically modified autologous stem cells. On the other hand, severe side effects in a few patients treated with gene therapy have also been reported, indicating the usefulness of further improving the vectors currently used in gene therapy clinical trials. In the field of stem cell biology, evidence showing the plastic potential of adult hematopoietic stem cells and data indicating the multipotency of adult mesenchymal precursor cells have been presented. Also, the generation of embryonic stem cells by means of nuclear transfer techniques has appeared as a new methodology with direct implications in gene therapy. PMID- 14734100 TI - Stem cell plasticity: the growing potential of cellular therapy. AB - The fundamental principle of stem cell biology is that cells with the potential for both self-renewal and terminal differentiation into one or more cell types may be found in a given tissue. The corollary of this principle is that the stem cells give rise to tissues in which they reside, the so-called expected tissues. Many exciting discoveries reported over the last several years challenge this paradigm by showing that there are not only tissue-specific stem cells that differentiate to the expected mature cells, but also that tissue stem cells can differentiate into unexpected cell lineages, suggesting an enormous plasticity of differentiation. Hematopoietic stem cells, which have drawn the most attention, mesenchymal stem cells, and neural stem cells have been the focus of many investigations. However, recent studies directed toward hematopoietic stem cells have disputed the concept of stem cell plasticity, suggesting that experimental artifact or somatic cell fusion may account for reported observations of plasticity. Although the data are mounting, stem cell plasticity, strictly defined, has yet to be rigorously proven. Animal models to meticulously define the biology and potential plasticity of stem cells and pilot clinical trials to begin to explore the biology and therapeutic potential of human stem cells will both be vital to advance the field over the coming years. PMID- 14734101 TI - Ethical and legal issues in therapeutic cloning and the study of stem cells. AB - Therapeutic cloning is a new technology with great medical potential, particularly in the area of transplantation medicine. It involves the transfer of the nucleus of a patient's cell into an enucleated donor oocyte for the purpose of generating an embryo. This embryo is allowed to grow until the blastocyst stage, at which time stem cells can be obtained and differentiated into the tissue needed. Stem cells can also be obtained from adult tissues, as they seem to have sufficient plasticity to use for the stated purpose. A literature review was performed, and it is clear that the main controversy regarding the use of stem cells is the origin. Few people would object to their use if obtained from adult tissues; however, many oppose harvesting them from embryos in the blastocyst stage regardless of whether 1) they are obtained from surplus embryos donated by couples after assisted reproductive techniques, or 2) they are specially manufactured for research purposes. The central reason is the consideration that embryos should be treated as full humans from the moment of fertilization. This argument is also at the bottom of an older discussion regarding the validity of abortion. There is no consensus at the present time in this regard, and it is unlikely one will be forthcoming in the future. Arguments on both sides of the issue are presented, but emphasis is made on the need for using this technology for research purposes because of its potential value as a therapeutic tool. PMID- 14734103 TI - Immunity in Caenorhabditis elegans. AB - Until very recently it was not known whether the invertebrate Caenorhabditis elegans was capable of mounting a specific immune response to protect itself from pathogens. It has only just become clear that this simple nematode in fact possesses a complex innate immune system, involving multiple signalling pathways and an armoury of antimicrobial proteins and peptides. Genetic and microarray approaches are now revealing the molecular cross-talk that exists between the different signalling cascades. PMID- 14734104 TI - Blood cells of Drosophila: cell lineages and role in host defence. AB - Drosophila haemopoiesis gives rise to three independent cell lineages: plasmatocytes, crystal cells and lamellocytes. The regulation of Drosophila stem cell proliferation and lineage specification involves transactivators and signalling pathways, many of which have mammalian counterparts that control haemopoietic processes. Drosophila plasmatocytes are professional phagocytes that resemble the monocyte/macrophage lineage, crystal cells play a critical role in defence-related melanisation, and lamellocytes encapsulate large invaders. Crystal cells and lamellocytes have no clear mammalian homologues. Research into the molecular mechanisms that underlie the various immune functions of Drosophila blood cells, such as non-self recognition, is now taking wing. PMID- 14734105 TI - Mosquito immune responses against malaria parasites. AB - Anopheline mosquitoes are the major vectors of human malaria. Mosquito-parasite interactions are a critical aspect of disease transmission and a potential target for malaria control. Mosquitoes vary in their innate ability to support development of the malaria parasite, but the molecular mechanisms that determine vector competence are poorly understood. This area of research has been revolutionized by recent advances in the mosquito genome characterization and by the development of new tools for functional gene analysis. PMID- 14734106 TI - Activation of dendritic cells: translating innate into adaptive immunity. AB - Innate recognition of infection in vertebrates can lead to the induction of adaptive immune responses through activation of dendritic cells (DCs). DCs are activated directly by conserved pathogen molecules and indirectly by inflammatory mediators produced by other cell types that recognise such molecules. In addition, it is likely that DCs are activated by poorly characterised cellular stress molecules and by disturbances in the internal milieu. The multiplicity of innate pathways for DC activation may have evolved to ensure that any signs of infection are detected early, before overwhelming pathogen replication. Understanding which of these signs are both necessary and sufficient to convert DCs into the immunostimulatory antigen-presenting cells that prime appropriate effector T cells may hold the key to improved strategies for vaccination and immunotherapy. PMID- 14734107 TI - Recognition of infected cells by natural killer cells. AB - Under the influence of cytokines associated with innate immunity, natural killer (NK) cells rapidly become activated and migrate to sites of infection. Upon contact with infected parenchyma they proliferate, release cytokines and/or kill cells harboring pathogens. Multiple stimulatory and inhibitory receptors can provide the integrated signals that trigger this contact-mediated NK-cell function. Recent work has begun to define the ligands for these receptors in the context of infection by certain well-studied viruses. These results, in addition to future work involving other pathogens, will provide an understanding of the molecules present on parasitized cells that mark them as targets of innate immunity. PMID- 14734108 TI - Inherited disorders of NF-kappaB-mediated immunity in man. AB - The transcription factors of the NF-kappaB family play an important role in immunity to infection in animal models. Three human primary immunodeficiencies associated with impaired NF-kappaB signaling were recently described. X-linked recessive anhidrotic ectodermal dysplasia with immunodeficiency (XL-EDA-ID) is caused by hypomorphic mutations in the gene encoding NEMO/IKKgamma, the regulatory subunit of the IkappaB-kinase (IKK) complex. Autosomal dominant EDA-ID (AD-EDA-ID) is caused by a hypermorphic mutation in the gene encoding the inhibitory protein IkappaBalpha. Autosomal recessive immunodeficiency without EDA is caused by mutations in the gene encoding IRAK-4, a kinase acting upstream from the IKK complex in the TIR signaling pathway. The description of the infectious phenotypes associated with these genetic defects has initiated the forward genetic dissection of NF-kappaB-mediated immunity in man. PMID- 14734109 TI - NADPH oxidase. AB - NADPH oxidase is an enzyme that catalyzes the production of superoxide from oxygen and NADPH. It is a complex enzyme consisting of two membrane-bound components and three components in the cytosol, plus rac 1 or rac 2. Activation of the oxidase involves the phosphorylation of one of the cytosolic components. Recent crystallography data indicate that the tail of this cytosolic component lies in a groove between two Src homology 3 domains and, when phosphorylated, the tail leaves the groove and is replaced by the tail of one of the membrane-bound components. Chronic granulomatous disease is an inherited immune deficiency caused by the absence of one of the components of the oxidase. The most important recent advances in the field have been the crystallographic analysis of the oxidase and the use of antifungal agents in the prophylaxis of chronic granulomatous disease. PMID- 14734110 TI - Plant innate immunity - direct and indirect recognition of general and specific pathogen-associated molecules. AB - Plants have the capacity to recognise and reject pathogens at various stages of their attempted colonisation of the plant. Non-specific rejection often arises as a consequence of the potential pathogen's attempt to breach the first lines of plant defence. Pathogens able to penetrate beyond this barrier of non-host resistance may seek a subtle and persuasive relationship with the plant. For some, this may be limited to molecular signals released outside the plant cell wall, but for others it includes penetration of the cell wall and the delivery of signal molecules to the plant cytosol. Direct or indirect recognition of these signals triggers host-specific resistance. Our understanding of host-specific resistance and its possible links to non-host-specific resistance has advanced significantly as more is discovered about the nature and function of the molecules underpinning both kinds of resistance. PMID- 14734112 TI - Transcriptional regulation of antigen presentation. AB - MHC class I and class II molecules play essential roles in the adaptive immune response by virtue of their ability to present peptides to T lymphocytes. Given their central role in adaptive immunity, the genes encoding these peptide presenting molecules are regulated in a tight fashion to meet with local requirements for an adequate immune response. In contrast to MHC class I gene products, which are expressed on almost all nucleated cells, constitutive expression of MHC class II molecules is found only in specialized antigen presenting cells of the immune system. Expression of both classes of MHC molecules can be induced by immune regulators and upon cell activation. A set of conserved cis-acting regulatory promoter elements mediate the transcription of MHC class I and beta2-microglobulin genes. Of these regulatory elements, the promoters of MHC class II and accessory genes also have the SXY module. The MHC class II transactivator (CIITA) is essential for the activation of MHC class II promoters, and it functions through protein-protein interactions with regulatory factors bound to the SXY module. Given the central role of CIITA in these regulatory processes, it is of interest to identify the DNA-binding factors and co-activators that assemble on CIITA promoters in a cell-type-specific fashion. Accordingly, recent studies include investigations into chromatin remodeling and epigenetic control mechanisms that modulate cell-type-specific transcriptional regulation of genes involved in antigen presentation. PMID- 14734113 TI - Proteasome and peptidase function in MHC-class-I-mediated antigen presentation. AB - MHC-class-I-presented peptides are predominantly generated by the proteasome system. IFN-gamma strongly influences the processing efficiency by inducing immunoproteasome formation and proteasome activator PA28 synthesis. Depending on the protein substrate, the presence of immunoproteasomes and PA28 influence epitope liberation either positively or negatively. Abundantly occurring defective ribosomal products are a major source for proteasome-dependent antigen processing; however, antigen presentation is relatively inefficient. This is in part due to the existence of a panel of cytosolic aminopeptidases, such as bleomycin hydrolase (BH), puromycin-sensitive aminopeptidase (PSA) and thimet oligoendopeptidase (TOP), that can destroy epitopes or their precursors. Other aminopeptidases, such as leucine aminopeptidase (LAP) and endoplasmic reticulum aminopeptidase 1 (ERAP 1), can trim epitope precursors from the amino terminus to their correct size for MHC class I binding to enhance antigen presentation. Recent evidence suggests that tripeptidyl peptidase II (TPPII), a large peptidase with exo-and endo-proteolytic activities, is also involved in antigen processing and may generate a specific set of MHC class I epitopes. PMID- 14734114 TI - Recent developments in MHC-class-I-mediated antigen presentation. AB - Defective ribosomal products provide an important supply of endogenous peptides for entry into the classic MHC class I antigen presentation pathway. The recruitment of endoplasmic reticulum membrane during phagosome biogenesis allows exogenous antigens to be translocated into the cytosol as well as providing access to the class I peptide transport and loading machinery. This combination of features provides a mechanism for cross-presentation by specialised antigen presenting cells. Recent studies have shed new light on these pathways and have also described the emerging K3 family of viral ubiquitin E3 ligases, which constitutively ubiquitinate and degrade MHC class I molecules and other immunoreceptors. PMID- 14734115 TI - New insights into pathways for CD1-mediated antigen presentation. AB - Recent studies of CD1 structure and intracellular trafficking have demonstrated significant differences among the CD1 isoforms (CD1a, CD1b, CD1c and CD1d). The molecular and structural basis for the differential trafficking of CD1 molecules has also been delineated. These observations broaden our understanding of why the immune system has evolved multiple CD1 isoforms to survey different cellular compartments for lipid antigen presentation, to provide host defense against the microbial world and to offer immunoregulation with relevance to tumor immunity and autoimmunity. PMID- 14734116 TI - Class II MHC peptide loading by the professionals. AB - The loading of class II MHC molecules with antigenic peptides is largely confined to the endocytic vesicles of specialized antigen-presenting cells (APCs), such as B cells, macrophages and dendritic cells. At first glance, the pathway utilized by each of these professional APCs to generate class II-peptide complexes on their surface appears to be indistinguishable. All three types of APC rely on the chaperone Ii for correct class II assembly and transport to the endocytic pathway, they all depend on the action of specific cysteine proteases to remove Ii from the class II-Ii complex, and they all utilize the class II-like molecule DM to facilitate peptide loading. A closer look, however, reveals subtle yet important differences in the class II maturation pathway between each of these APCs, which befit the unique roles these individual cells play in eliciting CD4(+) T-cell responses. PMID- 14734117 TI - Association of MHC class II-peptide complexes with plasma membrane lipid microdomains. AB - Activation of CD4(+) T cells requires the interaction of multiple T-cell receptors with MHC class II-peptide complexes on the surface of antigen presenting cells (APCs). Recent studies have shown that MHC class II complexes are clustered in APC plasma membrane microdomains, thereby providing a mechanism for localized concentration of MHC class II-peptide complexes. The integrity of one type of APC membrane microdomain, the lipid raft, is important for antigen presentation to T cells. We propose a model in which the coordinated processes of MHC class II peptide loading and intracellular trafficking enhance T-cell activation by loading specific MHC class II-peptide complexes in discrete lipid raft microdomains. PMID- 14734118 TI - MHC class II signaling in antigen-presenting cells. AB - The MHC class II molecules have been recognized as signaling receptors for more than a decade, and recent work has revealed the importance of their signaling for the immune response. Today, we know that the function of MHC class II molecules on antigen-presenting cells (APCs) is not limited to their role as antigen presenting structures; they are flexible receptors that, by triggering a variety of signaling pathways, can regulate APC activities from proliferation and maturation to apoptosis. Recent advances have provided insights into how these molecules might accommodate such regulation. PMID- 14734119 TI - Molecular interactions at the T cell-antigen-presenting cell interface. AB - The development of new imaging techniques has made it possible to investigate the dynamic movements of molecules involved in T-cell signalling. Fluorescence resonance energy transfer (FRET) imaging allows the investigation of protein protein interactions in live cells, and has demonstrated that T-cell receptors (TCRs) and CD4 are brought together in the immunological synapse during antigen recognition. This interaction is inhibited by antagonist ligands. Antagonism works through competition between agonist and antagonist ligands for TCR binding, as well as through feedback via the SHP-1 tyrosine phosphatase and extracellular signal-related kinase. Early signalling events result in the clustering of co receptors and TCRs at the synapse, and the activation of various signalling molecules. Recent data show that some T-cell signalling precedes the formation of the mature form of the immunological synapse, but that full T-cell activation depends on sustained signalling, which in turn requires the synapse. PMID- 14734120 TI - In vivo antigen presentation. AB - Monoclonal antibodies specific for defined peptide-MHC complexes are now being used to physically detect T-cell receptor ligands. These reagents have resulted in the identification of the cells that present antigen in lymphoid and non lymphoid tissues after various forms of antigen administration. In addition, recent advances in real-time imaging technology have begun to measure the rate and directionality of T-cell movement relative to antigen-presenting cells in lymph nodes, shedding light on the earliest events in T-cell activation in a physiological setting. PMID- 14734121 TI - Non-heart-beating donors in thoracic transplantation. AB - Access to lung transplantation is severely limited by a scarcity of suitable donors, resulting in increasing numbers of deaths on the heart and lung transplant waiting lists, and strict selection criteria for recipients. Unlike some other solid organs, the lung may be ideally suited to retrieval for transplant following substantial intervals after circulatory arrest. This may be because lung parenchymal cells do not rely on perfusion for cellular respiration. This review outlines the relevant published experimental data that addresses the concept that lungs might be suitable for transplant even if retrieved from non heart-beating donors (NHBDs), and the small published clinical experience with NHBDs as lung donors. Aspects of reperfusion injury in this setting are reviewed. The prospect of heart transplant from NHBDs is addressed. The impact of the routine use of NHBDs on lung transplantation is discussed. PMID- 14734122 TI - Effects of passive cardiac containment on left ventricular structure and function: verification by volume and flow measurements. AB - BACKGROUND: The cardiac support device (CSD, Acorn) is a compliant, textile-mesh graft placed around the ventricles to prevent further dilatation and to improve function in congestive heart failure. The aim of this study was to verify post operative changes in left ventricular volumes, ejection fraction, blood flow, and myocardial mass. METHODS: Fourteen patients underwent contrast-enhanced, electrocardiography-triggered electron-beam computerized tomography before and 6 to 9 months after CSD implantation. We measured volume and flow using the slice summation method and the indicator-dilution technique. RESULTS: We found significant changes for the following parameters: end-diastolic volume decreased from 382.9 +/- 140.2 ml to 311.3 +/- 138.7 ml, end-systolic volume from 310.4 +/- 132.4 ml to 237.4 +/- 133.8 ml, end-diastolic diameter from 75.3 +/- 7.8 mm to 70.7 +/- 11.6 mm, end-systolic diameter from 65.8 +/- 7.8 mm to 60.0 +/- 14.0 mm, and myocardial mass from 298.6 +/- 79.6 g to 263.1 +/- 76.8 g. Ejection fraction increased from 20.3% +/- 6.4% to 27.8% +/- 13.1%. We found no significant differences for stroke volume (from 72.5 +/- 24.6 ml to 73.8 +/- 23.6 ml), heart rate (from 80.5 +/- 11.0 beats per minute to 76.5 +/- 6.8 beats per minute), and total cardiac output (from 5.8 +/- 1.9 liter/min to 5.6 +/- 1.8 liter/min). Mitral regurgitation fraction decreased from 30.5% +/- 15.5% to 15.6% +/- 12.8%, increasing antegrade cardiac output from 3.8 +/- 0.9 liter/min to 4.7+/-1.5 liter/min. For most parameters, pre- and post-operative values in these patients differed significantly from those in an age- and gender-matched control group. In each patient, we observed a small hyperdense stripe along the pericardium after surgery, but we observed no local complications. CONCLUSION: Three-dimensional structural and functional data obtained by computerized tomography volume and flow measurements confirm the safety and efficacy of CSD implantation. PMID- 14734123 TI - Neurohumoral and hemodynamic effects of the selective endothelin antagonist darusentan in advanced chronic heart failure. AB - BACKGROUND: Endothelin antagonists represent a new approach to neurohumoral treatment in patients with chronic heart failure. In this study, the new selective endothelin-A receptor antagonist, darusentan, was compared with placebo for 3 weeks in patients with severe heart failure on top of standard treatment that included angiotensin-converting enzyme (ACE) inhibitors and beta-blockers. Effects on neurohormones and hemodynamics were evaluated. METHODS: Consecutive patients with severe heart failure (New York Heart Association [NYHA] Grade III) were included in this neurohumoral sub-study of an international, multi-center, double-blind, placebo-controlled study of darusentan, and randomized to darusentan (n = 23) or placebo (n = 8). The mean left ventricular ejection fraction was 19 +/- 6% at the beginning of the study. Patients were randomized to different dosage levels of darusentan (30, 100, or 300 mg) for 3 weeks. Hemodynamics were obtained by right heart Swan-Ganz catheterization at entry and end of study. Serial assessment of plasma brain natriuretic peptide (BNP), big endothelin, and pro-atrial natriuretic peptide (pro-ANP) was performed. In the active treatment group, 1 patient died due to worsening heart failure, 1 patient received elective heart transplantation, and 2 patients stopped taking the medication due to vertigo. In the placebo group, 1 patient was excluded due to non-compliance. RESULTS: Overall, the mean dose of darusentan was 144 +/- 125 mg/day (30 mg: n = 8; 100 mg: n = 4; 300 mg: n = 7). Significant benefits in hemodynamic variables were found after 3 weeks only in patients receiving darusentan (baseline vs end of study: cardiac index: 2.0 +/- 0.3 vs 2.6 +/- 0.5 liters/min m(2), p < 0.0001; mean pulmonary artery pressure: 35 +/- 9 vs 33 +/- 8 mm Hg, p < 0.05; heart rate: 79 +/- 16 vs 71 +/- 10 beats/min, p < 0.01). A significant reduction in mean arterial blood pressure was observed with the endothelin antagonist (baseline 80 +/- 8 vs end 73 +/- 8 mm Hg, p < 0.01). BNP decreased significantly in patients with darusentan (90 +/- 87 at entry vs 63 +/- 67 fmol/ml after 3 weeks, p < 0.01), whereas big-endothelin remained unchanged. Pro-ANP tended to decrease in the active treatment group, but did not reach statistical significance. CONCLUSION: Significant hemodynamic and neurohumoral benefits were observed in patients with severe heart failure receiving the selective endothelin antagonist darusentan. PMID- 14734124 TI - Evidence of improved right ventricular structure after LVAD support in patients with end-stage cardiomyopathy. AB - BACKGROUND: Although many reports demonstrate the hemodynamic benefits of left ventricular assist devices (LVAD) in right-sided circulation, it is not known whether the right ventricular myocardium goes through reverse remodeling after left ventricular mechanical circulatory support. Accordingly, the purposes of our studies were 1). to investigate the right ventricular changes that occur in fibrosis, in cellular hypertrophy, and in intra-myocardial tumor necrosis factor alpha (TNF-alpha) levels in patients receiving LVAD support; and 2). to determine whether the type of LVAD used influences right ventricular myocardial changes. METHODS AND RESULTS: We measured myocyte size, total collagen content, and TNF alpha levels using semi-quantitative immunohistochemical analysis of myocardial samples from the right and left ventricles of control and failing myocardia, either supported by 1 of 2 distinct forms of LVADs or without support. We found that when compared with control, although myocyte size was not increased in the right ventricle of failing myocardia (p = not significant), total collagen content and myocardial TNF-alpha levels were decreased in the right ventricle compared with controls (p < 0.01 and p < 0.001, respectively). CONCLUSION: These data demonstrate that chronic left ventricular unloading with either pulsatile or continuous-flow devices decreases right ventricular total collagen and myocardial TNF-alpha content. We suggest that the decreased fibrosis and normalization of cytokine milieu observed may in part contribute to the recovery of right-sided cardiac function associated with chronic mechanical circulatory support. PMID- 14734125 TI - Degree of cardiac fibrosis and hypertrophy at time of implantation predicts myocardial improvement during left ventricular assist device support. AB - BACKGROUND: There have been increasing reports of cardiac improvement in heart failure patients supported by left ventricular assist devices (LVADs i.e.), including a number of patients who have tolerated removal of the device without the benefit of cardiac transplant. In the current study, we retrospectively investigated echocardiographic and histologic changes in patients supported by LVADs (n = 18). The goal of our study was to determine if the degree of cardiac fibrosis and myocyte size in pre-implant biopsies could predict myocardial improvement as assessed by improvements in ejection fraction (EF) during LVAD support. METHODS: We determined total collagen content in myocardial biopsy specimens by a semi-quantitative analysis of positive Picro-Sirius Red-stained areas and myocyte size measurements by computerized edge detection software. RESULTS: During LVAD support, 9 of the 18 patients (Group A) were distinguished by significant improvement in ejection fraction (pre <20% vs unloaded 34 +/- 5%). In addition, Group A patients had significantly less fibrosis and smaller myocytes than their Group B counterparts, whose EF did not improve. There was an inverse correlation between pre-implant biopsy collagen levels and myocyte size with increases in EF during LVAD unloading. CONCLUSIONS: We found that the patients who demonstrated the greatest improvements in EF during support had less fibrosis and smaller myocytes at the time of device implantation. We propose that tissue profiling a patient's pre-implant biopsy for fibrosis and myocyte size may allow stratification in Stage IV heart failure and may predict myocardial improvement during LVAD support. PMID- 14734126 TI - Haptoglobin phenotype correlates with development of cardiac transplant vasculopathy. AB - OBJECTIVES: The purpose of this study was to investigate the association between haptoglobin phenotypic variation and development of cardiac transplant vasculopathy. BACKGROUND: The development of coronary vasculopathy determines long-term survival after cardiac transplantation. Serum haptoglobin levels are associated with non-transplant atherosclerosis. In addition to free hemoglobin binding, haptoglobin influences free radical formation, prostaglandin synthesis and angiogenesis. Three phenotypes of haptoglobin exist in humans, which have both quantitative and qualitative differences. METHODS: Coronary disease was diagnosed at post-transplant routine surveillance angiography. Hemoglobin (10%) was added to recipient plasma to form a haptoglobin-hemoglobin complex. Sample aliquots were applied to acid hemoglobin plates and electrophoretically separated. Phenotypes were recognized by comparing the electrophoretic pattern with that of established standards. Haptoglobin concentrations were measured using an immunoturbidimetric technique with polyethylene glycol (PEG)-enhanced precipitation. RESULTS: Ninety-three patients were independently studied. Phenotype 1-1 was found in 20.4%, 2-1 in 41.9% and 2-2 in 37.6%. Haptoglobin levels were highest in 1-1 recipients (2.1 +/- 0.58 g/liter) compared with 1.78 +/- 0.88 g/liter and 1.3 +/- 0.81 g/liter in 2-1 and 2-2 individuals, respectively (p = 0.001). Haptoglobin phenotype was significantly related to the development of vasculopathy; recipients with a 2-1 phenotype were more likely to develop angiographic disease (p = 0.0084). No differences were found among the 3 groups according to univariate analysis. Multivariate analysis identified 3 risk factors for vasculopathy development: age of donor (hazard ratio 1.056 [95% confidence interval 1.02 to 1.094], p = 0.0023); pre-transplant recipient body mass index (hazard ratio 1.116 [95% confidence interval 1.015 to 1.23], p = 0.024), and haptoglobin phenotype (hazard ratio 2.725 [95% confidence interval 1.031 to 7.19], p = 0.012). CONCLUSIONS: Haptoglobin, through phenotype-dependent mechanisms, correlates with the development of coronary vasculopathy. This finding furthers our understanding of the disease, opens up new areas of research, and may lead to novel therapies. PMID- 14734127 TI - Histomorphometric comparison of cardiac allograft vasculopathy in miniature swine. AB - BACKGROUND: Whether the pathologic characteristics of vascular lesions manifested in recipients with cardiac allograft vasculopathy (CAV) differ with the severity of the histocompatibility barrier crossed at transplantation or with the type or amount of immunosuppression used to prolong graft survival is unclear. We used miniature swine to determine whether a wide variance in heart transplantation protocols, both in histoincompatibility and immunosuppression, affects the histomorphometry of CAV. METHODS: We compared explanted hearts from major histocompatibility complex Class I-disparate recipients who were treated for 12 days with cyclosporine (Group 1) with minor-antigen-disparate hearts transplanted into mixed chimeric recipients previously engrafted with donor hematopoietic progenitor cells (Group 2). We analyzed coronary intimal lesions using computerized morphometry, immunohistochemistry, and TUNEL assay. Myocardial cytokine-gene expression was determined using RNAse protection assays and reverse transcriptase polymerase chain reaction. RESULTS: The prevalence of CAV in Group 2 was significantly less than that observed in Group 1, but the severity of the lesions in both groups was similar. The vascular lesions that developed in both groups demonstrated the presence of alpha-smooth-muscle-actin-positive spindle cells expanding the intima, with few inflammatory cells. We noted an absence of proliferating cell nuclear antigen activity and TUNEL-positive cells in both groups. We observed prominent myocardial interferon-gamma gene expression only in Group 1. CONCLUSION: Despite differences in myocardial interferon-gamma gene expression, the histology and severity of the vascular lesions in CAV did not vary significantly with different histoincompatibilities or treatment protocols. These results suggest that the origin of CAV cannot be determined by histology alone. PMID- 14734128 TI - Mycophenolate mofetil for secondary prevention of cardiac allograft vasculopathy: influence on inflammation and progression of intimal hyperplasia. AB - BACKGROUND: Cardiac allograft vasculopathy (CAV) remains the single most important complication impairing long-term survival after heart transplantation (HTx). Intimal hyperplasia as a response to immunologic and non-immunologic injury is involved in the pathogenesis. Because improved immunosuppressive properties with mycophenolate mofetil (MMF) have been shown within the first year, beneficial effects on intimal hyperplasia and systemic inflammation might be found late after HTx as well. METHODS: After a baseline examination with intravascular ultrasound (IVUS, volumetric assessment) 30 patients (2.0 +/- 1.1 years post-HTx) were prospectively randomized to receive either MMF (2 g/day) or to continue with azathioprine (AZA) as part of a triple immunosuppression protocol with cyclosporine and prednisolone. Markers of systemic inflammation and changes in vascular geometry were evaluated by IVUS after 1 year of follow-up. RESULTS: With regard to inflammation, significantly lower values were found for high-sensitive C-reactive protein (CRP) in the MMF group (AZA 1.8 +/- 1.2 mg/liter. vs MMF 1.0 +/- 4.1 mg/liter, p = 0.02). Tumor necrosis factor (TNF) alpha, interleukin (IL)-10, IL-6 and transforming growth factor (TGF)-beta did not differ between the groups. IVUS revealed no significant differences between groups. There was a weak trend toward a larger increase in plaque volume (AZA 13 +/- 43 mm(3) vs MMF 27 +/- 41 mm(3), p = 0.33), whereas MMF-treated patients tended to show a small increase in vessel dimensions (AZA +10 +/- 63 mm(3) vs MMF +50 +/- 87 mm(3), p = 0.17). CONCLUSIONS: Changing immunosuppression from a standard AZA-based regimen to MMF resulted in a decrease in systemic inflammatory activity as indicated by levels of high-sensitive CRP. However, progression of intimal hyperplasia did not differ significantly, and the weak trend toward vascular enlargement could indicate some influence on vascular geometry. PMID- 14734129 TI - Late pacemaker requirement after pediatric orthotopic heart transplantation may predict the presence of transplant coronary artery disease. AB - BACKGROUND: Few data are available regarding pacemaker implantation after pediatric orthotopic heart transplantation. The purpose of this study was to assess the incidence, indications and associations with regard to pacemaker placement in children who have undergone orthotopic heart transplantation. METHODS: We performed a retrospective study of all patients undergoing orthotopic heart transplantation at our institution from October 1984 to March 2001. Data obtained included demographics, indications for pacemaker, presence of transplant coronary artery disease and long-term follow-up. Patients were divided into: Group 1, patients requiring a pacemaker within 3 months of transplantation; and Group 2, patients requiring a pacemaker beyond 3 months. RESULTS: Pacemakers were required in 7 of 106 (6.6%) transplant recipients. Pacing indications for patients in Group 1 (n = 2) were persistent bradycardia with pause-related ventricular arrhythmia and atrial flutter with resultant sinus pauses of up to 4 seconds. In Group 2 patients (n = 5), indications for pacing were high-grade atrioventricular (AV) block in 1 patient and episodic sinus pauses up to 3.3 seconds associated with syncope/dizziness in the remaining 4 patients. All patients in Group 2 had transplant coronary disease diagnosed within 1 year of pacemaker implantation. All had resolution of symptoms and no complications after implantation. CONCLUSIONS: Pacemakers are infrequently required after cardiac transplantation in children. Despite not meeting classic symptomatic sinus bradycardia criteria, pacemaker placement should be considered post transplantation in patients with episodic sinus pauses and dizziness or syncope. Patients who present with the aforementioned symptoms or high-grade AV block should be evaluated closely for the presence or development of transplant coronary artery disease, as it may be their first symptom. PMID- 14734130 TI - Mitochondrial respiratory abnormalities in patients with end-stage congenital heart disease. AB - BACKGROUND: Nitric oxide (NO) binds to mitochondrial cytochrome oxidase to decrease myocardial oxygen consumption (MVO(2)). This regulation is disrupted in heart failure (HF) due to reduced NO. The present objective was to evaluate NO mediated regulation of mitochondrial respiration in the myocardium of patients with congenital heart disease (CHD) and cardiomyopathy (CMP). METHODS: MVO(2) was measured in vitro in explanted human myocardium obtained at transplantation. Seven patients had CHD (5 cyanotic, 2 acyanotic), and 11 had non-ischemic CMP. The effects of the following on MVO(2) were measured: kinin-dependent endothelial NO synthase (eNOS) agonists, bradykinin, ramiprilat and amlodipine; NO donors, nitroglycerin and S-nitroso-N-acetylpenicillamine (SNAP) (10(-7) to 10(-4) mol/liter); and NOS inhibitor, N(omega)-nitro-L-arginine methylester (L-NAME). RESULTS: eNOS agonists caused a smaller decrease in MVO(2) in CHD compared with CMP patients. Changes in MVO(2) at the highest dose in CHD vs CMP were, respectively: bradykinin, -22 +/- 7% vs: -30 +/- 5% (p < 0.05); ramiprilat, -17 +/- 8% vs -26 +/- 2%, (p < 0.001); and amlodipine, -5 +/- 7% vs -29 +/- 6% (p < 0.001). L-NAME attenuated the effect of bradykinin, ramiprilat and amlodipine in both groups, confirming that the drug effect was secondary to eNOS activation. Nitroglycerin and SNAP also caused smaller decreases in MVO(2) in CHD vs CMP (NTG -16 +/- 6% vs -37 +/- 4%, SNAP -37 +/- 4% vs -49 +/- 3%, [p < 0.01]), suggesting altered mitochondrial function in CHD. CONCLUSIONS: Abnormal regulation of MVO(2) in end-stage CMP may be secondary to reduced endogenous NO availability and can be reversed by the use of NO agonists. In end-stage CHD, this abnormality may be related in part to abnormal mitochondrial function. PMID- 14734131 TI - Pro-brain natriuretic peptide as marker of cardiovascular or pulmonary causes of dyspnea in patients with terminal parenchymal lung disease. AB - BACKGROUND: Increased plasma concentrations of pro-atrial natriuretic peptide (proANP) and pro-brain natriuretic peptide (proBNP) are features of left ventricular impairment. However, concentrations of proANP and proBNP in patients with isolated terminal parenchymal lung disease are not known. Therefore, we measured the plasma concentrations of natriuretic precursor peptides in patients with terminal parenchymal lung disease who had normal left ventricular function and who were referred for evaluation for lung transplantation. METHODS: We measured plasma N-terminal proANP and proBNP in patients undergoing right heart catheterization (n = 50) and related results to hemodynamic variables obtained during catheterization. RESULTS: Plasma proBNP concentrations were unaffected in patients with terminal parenchymal lung disease and normal left ventricular function (median, 2.5 pmol/liter; range, 0-22; upper reference limit, 15 pmol/liter). In contrast, patients with primary pulmonary hypertension displayed more than a 40-fold increase in plasma proBNP concentrations (median, 107 pmol/liter vs 2.5 pmol/liter, p < 0.0001). Plasma N-terminal proANP increased moderately (median, 664 pmol/liter; range, 36-1620; upper reference limit, 600 pmol/liter) but correlated to plasma proBNP concentrations (r = 0.47, p < 0.0001). Finally, regional vascular proBNP concentrations revealed the heart as the secretory site. CONCLUSIONS: Our findings strongly support the contention that natriuretic peptide measurements are efficient markers for cardiovascular causes of dyspnea. Moreover, our results eliminate natriuretic peptides as markers of moderate pulmonary hypertension in patients with terminal parenchymal lung disease. PMID- 14734132 TI - Diagnosis of pulmonary vascular limit to exercise by cardiopulmonary exercise testing. AB - BACKGROUND: Given the recent development of newer and less-invasive treatments for pulmonary hypertension, and the long wait for lung transplantation, early and correct diagnosis of this condition is increasingly important. The purpose of this study was to determine and improve the accuracy of a non-invasive, cardiopulmonary exercise-testing algorithm for detecting a pulmonary vascular limit to exercise. METHODS: We performed 130 consecutive, incremental cycling exercise tests for exertional symptoms with pulmonary and radial artery catheters in place. Pulmonary vascular limit was defined as pulmonary vascular resistance at maximum exercise >120 dynes. sec/cm(5) and a peak-exercise systemic oxygen delivery <80% predicted, without a pulmonary mechanical limit or poor effort. We applied a previously reported non-invasive exercise-test-interpretation algorithm to each patient and sequentially manipulated branch point threshold values to maximize accuracy. RESULTS: The sensitivity of the original non-invasive algorithm for pulmonary vascular limit was 79%, specificity was 75%, and accuracy was 76%. Sensitivity did not change with systematic alteration of branch-point threshold values, but specificity and accuracy improved to 88% and 85%, respectively. Accuracy improved most by modifying the threshold values for percent predicted maximum oxygen uptake and carbon dioxide output ventilatory equivalents at lactate threshold. CONCLUSION: Non-invasive cardiopulmonary exercise testing is a useful tool for detecting and excluding a pulmonary vascular limit and for determining whether abnormal pulmonary hemodynamics limit aerobic capacity. PMID- 14734133 TI - Risk quantification of early outcome after lung transplantation: donor, recipient, operative, and post-transplant parameters. AB - BACKGROUND: Because there is no reliable evaluation system of recipient acuity after lung transplantation, comparing patients among centers is difficult. The purpose of our study was to identify risk factors for 30-day mortality and prolonged intensive care unit stay and to develop a scoring system to evaluate the severity of impairment and to predict surgical outcomes. METHODS: We prospectively collected data from 122 lung transplant recipients and from 119 donors from January 1997 to June 2000. We assessed donor, recipient, and operative factors; ischemic time; and immediate post-operative physiologic parameters to identify risk factors for 30-day mortality and prolonged intensive care unit stay. Furthermore, we sub-classified these factors into grades to develop a scoring system for predicting surgical outcomes. RESULTS: Cardiopulmonary bypass use, body mass index >25 kg/m2, immediate post-operative systolic pulmonary arterial pressure, trend of oxygenation index from 12 to 24 hours after transplantation, and the Acute Physiology and Chronic Health Evaluation II score correlated significantly with outcomes, and the sum of these 5 scores correlated strongly with outcomes (p < 0.0001). CONCLUSIONS: We conclude that the total score of these 5 risk factors could be used to predict 30-day mortality and prolonged intensive care unit stay. This scoring system also will facilitate standardization among transplant centers in evaluating post-transplant severity of illness. PMID- 14734134 TI - Phrenic nerve dysfunction after heart-lung and lung transplantation. AB - BACKGROUND: Phrenic nerve dysfunction (PND) is a well-known complication after cardiac surgery, but reports on its incidence and consequences after heart-lung and lung transplantation are scarce. METHODS: The incidence and consequences (ventilator days and intensive-care unit length of stay [ICU LOS]) of PND were studied by retrospective chart review of 27 heart-lung (HLTx) and 111 lung (LTx) transplantations performed from July 1991 to June 2001 at the Leuven University Hospital, Leuven, Belgium. On clinical suspicion of diaphragmatic dysfunction, nerve conduction studies were performed, which were completed with a needle electromyogram (EMG) of the diaphragm when the conduction study was non conclusive. RESULTS: The incidence of PND in 21 evaluable HLTx recipients was 42.8% (9 of 21 patients), resulting in significantly more ventilator days for PND patients (37.6 +/- 36.3 days vs 5.3 +/- 3 days; p < 0.05) and a prolonged ICU LOS (46.8 +/- 33 vs 9.8 +/- 4.9 days; p < 0.05). In the 97 evaluable LTx patients, 9.3% (9 of 97 patients) developed PND. This resulted in more ventilator days for the PND group (30.6 +/- 14.8 days vs non-PND 7.9 +/- 14.8 days. p < 0.05) and a longer ICU LOS (PND 37.8 +/- 18.7 days vs non-PND 12.1 +/- 17.8 p < 0.05). Needle EMG of the diaphragm revealed denervation in 1 HLTx and 5 LTx patients. In LTx patients sustaining PND more tracheostomies were performed (44.4% vs 4.5% for non PND patients p < 0.005). Eight of 9 LTx patients with PND had sequential single lung transplantation. CONCLUSIONS: PND represents an important clinical problem after HLTx and LTx and has a considerable influence on both number of ventilator days and ICU resource utilization. PMID- 14734135 TI - Standardized guidelines for surveillance bronchoscopy reduce complications in lung transplant recipients. AB - BACKGROUND: The role of surveillance bronchoscopy in the care of lung transplant recipients remains controversial. Although there are no controlled studies to suggest a survival advantage, many transplant physicians support the practice. The procedure is generally safe but is associated with some complications. A review of practices at our institution revealed significant variation in patient preparation, management of risk related to the procedure, and in the technical aspects of the bronchoscopy itself. In an effort to minimize these differences and potentially improve outcomes, a standard set of procedural guidelines for all bronchoscopies was adopted in January 2000. METHODS: Reports from 1028 surveillance bronchoscopies performed in our outpatient facility from January 1999 to December 2001 were reviewed. Baseline patient data and procedure-related complications were identified. Specific complications recorded included oversedation, the need for prolonged supplemental oxygen, major and minor bleeding, pneumothorax, bronchospasm, vomiting, arrhythmia, hypotension and death. Differences between groups were analyzed using chi-square or Student's t tests as appropriate. RESULTS: The incidence of complications after the introduction of the guidelines (2000 and 2001) was significantly lower than in the year prior (1999) (1.95% vs 6.45%, p < 0.001). The lower rate of adverse events was mainly a result of a reduction in the incidence of minor bleeding (0.28% vs 2.26% p = 0.006) and of sedation-related complications (0.97% vs 2.90%, p = 0.04). CONCLUSIONS: The use of a standardized set of guidelines for surveillance fiber-optic bronchoscopy reduces complication rates. Similar guidelines should be considered by transplant centers performing the procedure. PMID- 14734136 TI - The value of ventilation scintigraphy after single lung transplantation. AB - BACKGROUND: A decrease in forced expiratory volume in 1 second (FEV(1)) as a diagnostic criterion for bronchiolitis obliterans syndrome (BOS) after single lung transplantation may be influenced significantly by the presence of the native lung. To quantify and to discriminate between the relative contribution of graft and native lung to the FEV(1), we retrospectively investigated the diagnostic value of combined FEV(1) measurements and ventilation scintigraphy in pulmonary dysfunction after single lung transplantation in 11 recipients with pulmonary vascular disease, 3 with obstructive lung disease, and 3 with restrictive lung disease. METHODS: We assessed function of the native lung and the graft, and subsequently calculated an adjusted grading of BOS by correcting routine FEV(1) measurements using linear interpolation of bi-annual lung ventilation scans. RESULTS: The contribution of the native lung to the total FEV(1) was slight (median, 9%) in recipients with obstructive disease compared with recipients with vascular (38%) or restrictive lung diseases (27%). Adjusted BOS grading was not useful in patients with obstructive disease. In the other patient groups, the onset of adjusted BOS Grade 1 and standard BOS Grade 1 was at a median of 220 days (range, 127-1146 days) and 836 days (184-3065 days), respectively. CONCLUSION: Ventilation scintigraphy is a useful adjunct in the (early) diagnosis of BOS in recipients of single lung transplants who have vascular and restrictive lung diseases. PMID- 14734137 TI - N-acetylcysteine inhibits interleukin-17-induced interleukin-8 production from human airway smooth muscle cells: a possible role for anti-oxidative treatment in chronic lung rejection? AB - BACKGROUND: Long-term survival of lung transplantation is threatened by obliterative bronchiolitis, or its clinical equivalent, bronchiolitis obliterans syndrome. With a prevalence of >50% at 5 years after transplantation, it has emerged as the most significant long-term complication. Neutrophilic inflammation and increased interleukin (IL)-8 production seem to be part of the basic pathophysiologic mechanism of chronic rejection. Recently, it has been suggested that reactive oxygen species may also play an important role in the pathogenesis because they are known to induce smooth muscle proliferation. METHODS: Human airway smooth muscle cells in vitro were stimulated with IL-17 (0.1 to 10 ng/ml) or with IL-17 (10 ng/ml) and the anti-oxidative agent N-acetylcysteine (1 micromol/liter to 10 mmol/liter). Production of 8-isoprostane was measured with a commercially available enzyme immunoassay kit and production of IL-8 was measured using a standard enzyme-linked immunoassay technique. RESULTS: IL-17 produced a concentration-dependent increase in 8-isoprostane with a maximum of 136.5 +/- 15.5 pg/ml with IL-17 (10 ng/ml, p < 0.001, n = 12, vs unstimulated cells). N acetylcysteine (NAC) was able to decrease IL-17-induced 8-isoprostane production, with a maximum decrease of 59.3 +/- 9% (p < 0.001, n = 12) with 10 mmol/liter of N-acetylcysteine, which also decreased IL-17-induced IL-8 production in a concentration-dependent manner (with maximum inhibition of 86.3% when combined with NAC 10 mmol/liter as compared with IL-17 alone). CONCLUSIONS: We demonstrated that human airway smooth muscle cells, when stimulated with IL-17, are able to produce 8-isoprostane, which could be inhibited by adding N acetylcysteline, and which was also able to decrease IL-17-induced IL-8 production. The clinical significance of these in vitro findings for prevention or treatment of chronic rejection after lung transplantation remains to be investigated. PMID- 14734138 TI - Novel broad-spectrum chemokine inhibitor protects against lung ischemia reperfusion injury. AB - BACKGROUND: Functional roles for chemokines have been demonstrated in several models of ischemia-reperfusion injury. The redundancy inherent to chemokine pathways makes administration of neutralizing antibodies to any single chemokine ineffective in ameliorating injury. This study was undertaken to define the pattern of chemokine expression in lung ischemia-reperfusion injury (LIRI), and to determine whether a broad-spectrum chemokine inhibitor, NR58-3.14.3, could confer significant protection against LIRI. METHODS: Left lungs of rats were rendered ischemic for 90 minutes and then reperfused for 4 hours. Chemokine secretion into the alveolar space was quantified by enzyme-linked immunoassay. Treated animals received NR58-3.14.3 prior to reperfusion. Vascular injury was measured by lung permeability index, neutrophil accumulation in lung parenchyma was determined by myeloperoxidase (MPO) activity, and alveolar leukocyte counts were quantified in bronchoalveolar lavage (BAL) effluent. A ribonuclease protection assay evaluated mRNA expression of various chemokines. RESULTS: Lavage effluent in untreated animals demonstrated significant increases in the secretion of cytokine-induced neutrophil chemoattractant (CINC), tumor necrosis factor alpha (TNF-alpha), macrophage inflammatory protein-2 (MIP-2), MIP-1alpha and monocyte chemoattractant protein-1 (MCP-1). Animals receiving NR58-3.14.3 demonstrated a 37% (p < 0.001) reduction in vascular injury and a marked reduction in lung MPO activity (p < 0.001) and alveolar cell counts (p = 0.005). Chemokine inhibition decreased mRNA expression of a number of early response cytokines when compared with positive control animals, and caused a significant decrease (p < 0.04) in the secretion of TNF-alpha. CONCLUSIONS: These studies demonstrate that chemokines are expressed after lung ischemia and reperfusion, and that broad-spectrum chemokine inhibition ameliorates reperfusion injury. mRNA expression of early response cytokines was modulated, and the secretion of TNF alpha was decreased. PMID- 14734139 TI - Cytopathology of pulmonary alveolar proteinosis complicating lung transplantation. AB - Pulmonary alveolar proteinosis is a disorder of unknown origin that occurs rarely after lung transplantation. We identified a patient with pulmonary alveolar proteinosis 66 days after undergoing single lung transplantation for idiopathic pulmonary fibrosis. We based the diagnosis on the presence of amorphous clumps or globules of acellular and finely granular material in bronchoalveolar lavage fluid (BALF). This material persisted for an 18.5-month period and was present in 9 of 14 lavage specimens. However, despite its presence in the native lung at autopsy, the material was seen in only 1 of 14 transbronchial lung biopsy specimens. Although uncommon, pulmonary alveolar proteinosis can be diagnosed readily in BALF by its distinctive cytopathologic features and should be considered in the differential diagnosis of pulmonary disease in lung transplant recipients. PMID- 14734140 TI - Severe left ventricular failure after double lung transplantation: pathophysiology and management. AB - Patients undergoing bilateral lung transplantation for end-stage pulmonary hypertension may experience various complications. We describe a patient who underwent transplantation for chronic pigeon breeder's disease, who had secondary pulmonary hypertension and deteriorated right heart function, and who developed severe left heart failure during the weaning phase after successful double lung transplantation. The patient was stabilized with catecholamines and an intra aortic balloon pump. Left heart function increased within 7 days and normalized at Day 18. Otherwise, the post-operative course was uneventful. PMID- 14734142 TI - Intractable ventricular tachycardia and bridging to heart transplantation with a non-pulsatile flow assist device in a patient with isolated left-ventricular non compaction. AB - Intractable ventricular tachycardia was investigated in a 51-year-old man with isolated left ventricular non-compaction during implantation of an automated internal cardioverter-defibrillator. Favorable bridging to cardiac transplantation was achieved with the DeBakey left ventricular assist device (LVAD). PMID- 14734141 TI - Histology of in-stent restenosis in a transplanted heart. AB - We report the case of a 32-year-old heart transplant recipient who suffered several episodes of early Grade IIIA rejection and, on serial coronary angiography, demonstrated aggressive allograft vasculopathy. Severe lesions in the right coronary and left obtuse marginal arteries were stented; however, the disease was progressive and, almost 6 years after heart transplantation, she was retransplanted, providing us with the opportunity to examine the histology of transplant in-stent restenosis. She remains well 1 year after a second heart transplant operation. PMID- 14734143 TI - Use of an arteriovenous fistula to make a small patient "appear larger" to her LVAD. PMID- 14734144 TI - CYP2E1: from ASH to NASH. AB - The pathology of the liver in alcoholic steatosis and alcoholic steatohepatitis (ASH) is remarkably similar to that of nonalcoholic fatty liver disease (NAFLD), including nonalcoholic steatohepatitis (NASH), suggesting some common pathogenic mechanism. Studies carried out over the last three decades of possible mechanisms involved revealed one common link, namely the induction of cytochrome P4502E1. Its substrates include fatty acids, ketones and ethanol. These substances, when present chronically in large amounts, induce the activity of the enzyme which thereby contributes to the disposition of these substrates. This reaction, however, is associated with the release of free radicals which can cause lipid peroxidation and liver injury, including mitochondrial damage. Mitochondrial damage in turn exacerbates the oxidative stress. CYP2E1 can also convert various xenobiotics to toxic metabolites. When unchecked, this toxicity eventually results in inflammation and fibrosis, culminating in cirrhosis. Prevention of this disorder is based on limiting the substrates that induce the system, such as excessive fatty acid associated with obesity and excessive alcohol consumption. No effective pharmacologic treatment is presently available but there is ongoing research on possible inhibitors of CYP2E1, innocuous enough to be suitable for chronic human consumption and sufficiently effective to attenuate the CYP2E1 induction to avoid the consequences of its excessive activity while maintaining its physiologic role. PMID- 14734145 TI - Cellular immune response in primary biliary cirrhosis. AB - The generation of immune responsiveness to self-antigen can result in the pathogenic autoimmune damage of tissues mediated by both humoral and cellular immune responses. Primary biliary cirrhosis (PBC) constitutes a model of autoimmune disease reflective of other organ-specific autoimmune feature. Although the etiology of PBC remained elusive, growing data suggest the role of cell-mediated immune response in the pathogenesis of PBC. Indeed, autoreactive CD4 as well as CD8 T cells have been characterized and their epitopes defined. Molecular mimicry is implicated in the initiation of these autoreactive T cell responses. Moreover, selective enrichment of NKT cells in the liver of PBC is demonstrated using CD1d tetramer.In this review, we shall focus on the recent advance of cell-mediated immune responses in PBC, which may be directly associated with inflammatory response in PBC. PMID- 14734146 TI - Common pathogenic mechanisms in ASH and NASH. PMID- 14734147 TI - Report of the 15th follow-up survey of primary liver cancer. AB - In the 15th Nationwide follow-up survey of primary liver cancer, 18,843 newly registered patients (1998-1999) and 18,405 follow-up patients from 791 hospitals in Japan were analyzed. Of the newly registered patients, approximately 95% were patients with hepatocellular carcinoma (HCC) and 3.3% had intrahepatic cholangiocarcinoma (ICC). The patients were assessed using 194 items that were related to epidemiological and clinicopathological factors, diagnosis, and treatment. Furthermore, the survival rates of all of the newly registered patients in the 10th-15th follow-up survey were calculated for each histological type, background factor(s) and treatment, respectively. In patients with hepatocellular carcinoma, the survival rates of patients who underwent hepatectomy, ethanol injection therapy, microwave coagulation therapy, or transcatheter arterial embolization were calculated by tumor size, tumor number, and clinical stage. This follow-up survey will be helpful to assess the progress of research and medical practice in the treatment of primary liver cancer. PMID- 14734148 TI - Analysis of approach to therapy for chronic liver disease in an HCV hyperendemic area of Japan. AB - In 1990, we conducted an epidemiological study of 509 residents in a hepatitis C virus (HCV) hyperendemic area in Japan. The purpose of the present study was to examine the approach to therapy for liver diseases accompanied by HCV and hepatitis B virus (HBV) infections among the surviving residents in the town after 12 years. Fifty-three with HCV or HBV infections among 385 people who resided in the town were clearly analyzed in 2002. The number of persons diagnosed with the liver diseases was as follows: HCV-related asymptomatic healthy carrier (1), past history of HCV infection (15), chronic hepatitis C (22), HCV-related liver cirrhosis (6), HCV-related hepatocellular carcinoma (HCC) (5), and HBV-related asymptomatic healthy carrier (4). HCC was detected in residents who did not have periodic regular hospital checkups. Only 19% of the 53 inhabitants consulted liver medical specialists, and 75% (3/4) who received interferon therapy received treatment from a liver medical specialist. It is necessary to provide continuous medical treatment to HCV carriers, minimizing difference in treatment quality in different medical institution. An efficient HCV medical checkup and a program of subsequence health management are important problems to be solved for improved health care. PMID- 14734149 TI - Efficacy of early retreatment with interferon beta for relapse in patients with genotype Ib chronic hepatitis C. AB - BACKGROUND: Interferon (IFN) retreatment for hepatitis C virus (HCV) relapsers has been effective under some conditions. We conducted a randomized, controlled trial of IFN beta retreatment for HCV relapsers after IFN alpha. PATIENTS AND METHODS: We gave IFN beta 6MIU therapy to 43 patients who had relapse of HCV after the 24 weeks IFN alpha monotherapy. The 43 patients were randomly assigned to two groups: Group A started retreatment within 4 weeks after relapse; and Group B started retreatment 24 weeks or more after relapse. RESULTS: Nine patients showed sustained virological response (SR) to the retreatment. All of these patients were in a low viral load subgroup. The SR rate in Group A (8/22, 36%) was significantly higher than in Group B (1/21, 5%) (P=0.0128). Among patients with lower viral load, the SR rate in Group A (8/10, 80%) was also significantly higher than in Group B (1/8, 13%) (P=0.0076). CONCLUSION: The retreatment with IFN beta is effective for patients with HCV low viral load, and the sooner after the relapse the retreatment is started, the better the clinical results will be. PMID- 14734150 TI - A comparative study of the microcirculatory changes in the developing liver cirrhosis between the central and peripheral parts of the main lobe in mice. AB - A comparative study of the microcirculatory changes in the developing liver cirrhosis between the central and peripheral parts of the main lobe in mice. INTRODUCTION: To investigate whether a difference in microvascular vascular changes occurring in the peripheral and the central area of the main liver lobe exists and if, it can explain the liver atrophy observed predominantly in the peripheral part of the main liver lobe. MATERIALS AND METHODS: Liver cirrhosis was induced in mice and the microvascular changes asserted using an intravital microscopy system after 1, 2, 3 and 8 weeks of treatment. RESULTS: The mean blood velocity increased in the sinusoids; intrahepatic shunts appeared predominantly in the peripheral area and larger sized afferent terminal portal venules were rarefied in the peripheral area, with the central area being not affected. DISCUSSION: The difference between the peripheral and central area shows a decreased direct and indirect reduction of the blood flow to the periphery of the main lobe, through the reduced number of afferent terminal portal venules and the development of intrahepatic shunting. PMID- 14734151 TI - Reduced activity of matrix metalloproteinase-9 in trypanosoma cruzi-infected mouse embryo hepatocyte cell. AB - In this work, we are reporting differences in the proteolytic profile of Trypanosoma cruzi-infected and non-infected primary cultures of mouse embryo hepatocyte cells. In gelatin-SDS-PAGE, ours results showed the presence of a 100kDa metalloproteinase in the supernatant and in the cells of both systems and an 85kDa extracellular metalloproteinase found only in the non-infected hepatocyte cultures. An enzymatic assay using gelatin as substrate showed a decrease of 74 and 70% in metalloproteinase activity in the culture supernatant and in the cell hepatocyte system infected with T. cruzi, respectively. Western blotting analysis using anti-matrix metalloproteinase-9 (MMP-9) antibody recognized the 100 and 85kDa protein bands, indicating that hepatocyte metalloproteinases correspond to the latent and active forms of the gelatinase MMP-9, respectively. The localization of MMP-9 was established by immunocytochemistry analysis in the cytoplasm of the non-infected and infected hepatocyte cells. In normal and infected hepatocyte cells, cysteine-proteinases migrating in gelatin-SDS-PAGE at 60kDa were detected and should correspond to lysosomal cysteine-proteinases of T. cruzi (cruzipain) and hepatocytes. In T. cruzi-infected hepatocytes an increase of approximately 50% in this enzymatic activity was observed, possibly due to parasite's cruzipain. PMID- 14734152 TI - Inherited bleeding disorders: disorders of platelet adhesion and aggregation. AB - Platelet aggregation at sites of vascular injury is essential for the formation of the primary haemostatic plug. The mechanism of platelet aggregation under conditions of physiological flow is a complex multistep process, which requires the synergistic action of several different platelet receptors. Platelet interaction with collagen at sites of damage to the vascular endothelium involves adhesion, activation, secretion of platelet granular contents and finally aggregation. Other agonists other than collagen, such as fibrinogen, vWF and soluble agonists released from activated platelets (thromboxane A2 (TXA2) and ADP) are involved in platelet aggregation. Platelets express a variety of receptors including GP Ib-IX-V, GP VI, GP Ia-IIa and GP IIb-IIIa. One aspect of this complexity of function is the variety of inherited defects of platelet function. Hereditary disorders of platelet adhesion are Bernard-Soulier syndrome and von Willebrand disease. Glanzmann thrombasthenia is an inherited disorder of platelet aggregation. The application of molecular biology to the study of platelet disorders has identified defects in other collagen receptors, ADP receptors and TXA2 receptors. Defects affecting TXA2 production, the generation of procoagulant activity and secretion from dense bodies and alpha-granules are also encountered. Other rare diseases, Chediak-Higashi, Hermansky-Pudlak and Wiskott-Aldrich syndrome also affect platelet storage granules. In this article, recent advances in the understanding of platelet function and knowledge of inherited disorders that affect platelet adhesion and aggregation is reviewed. As progress advances towards individualisation of therapy the phenotypic bleeding tendency of each patient becomes relevant. PMID- 14734153 TI - The importance of parenting concerns among patients with cancer. AB - Almost one quarter of all cancer patients in the United States is estimated to have children under 18-years-old living with them. However, there is limited research on how children's functioning is affected by parental cancer, and the impact of parenting concerns on treatment decisions. This paper will review the literature and hypothesize about why addressing the needs of the children has not been part of routine cancer care. In addition, we will make a case for the importance of including parenting issues in cancer care, and in assessments of treatment decision-making and quality of life. We also describe a clinical program that is attempting to address these issues, and offer practical recommendations for oncologists who want to help parents with cancer and their partners support their children through the challenges of cancer treatment. PMID- 14734154 TI - Health cooperation in an hospital of a rural area of Ivory Coast: analysis of the priorities and of the different levels at which cooperation can take place. AB - This paper critically reviews an experience of health cooperation in an hospital of a rural area of Ivory Coast. This particular situation is analysed in the more general frame of health problems in low-income countries and may suggest priorities for international health cooperation. The analysis of the main causes of avoidable death in poor countries does indicate targets and tools of intervention. In this case, the target was the reduction of infant mortality from anaemia of different origin and from HIV-1 mother-to-infant transmission. The major tool for intervention was the partnership between an Italian teaching and research hospital and the African hospital, with the catalyst of a non governmental organisation. This paper analyses the different levels at which cooperation developed in this project, from sheer economic support to the implementation of disease-oriented twinning programs that can improve health care and strengthen research capacity on both sides. Besides, medical, ethical and social implications of the ongoing cooperation program are discussed, with particular reference to the problems of preventing mortality from severe anaemia (diet fortification in children and pregnancy and transfusional guidelines in severe malaria) and of preventing mother-to-child neonatal transmission of HIV-1 infection (counselling and testing pregnant women for HIV-1, nevirapine administering to the mother and the baby and breast-feeding). PMID- 14734155 TI - Utilization of comprehensive geriatric assessment in cancer patients. AB - A growing and diverse aging population, recent advances in research on aging and cancer, and the fact that a disproportional burden of cancer still occurs in people aged 65 years and older have generated great interest in delivering better cancer care for older adults. This is particularly true as more survivors of cancer live to experience cancer as a chronic disease. Cancer and its treatment precipitate classic geriatric syndromes such as falls, malnutrition, delirium, and urinary incontinence. Comprehensive Geriatric Assessment (CGA), by taking all patient's needs into account and by incorporating patient's wishes for the level of aggressiveness of treatment, offers a model of integrating medical care with social support services. It holds the promise of controlling health care costs while improving quality of care by providing a better match of services to patient needs. Three decades after the CGA was initially developed in England, oncologists have begun taking notice on the potential benefits that CGA might bring to the field of geriatric oncology. This article describes the utilization of the CGA in cancer patients with an eye toward promoting interdisciplinary care for older cancer patients. To set an initial context, a search of computerized databases took place, using "comprehensive geriatric assessment" and "cancer" as keywords. A selection of literature from between 1980 and 2003 was reviewed. Additional articles were identified through the bibliography of relevant articles. PMID- 14734156 TI - A comprehensive geriatric intervention detects multiple problems in older breast cancer patients. AB - Studies of comprehensive geriatric assessment (CGA) have shown the importance of follow-up for effectiveness, but this has not been tested in an oncology clinic. In this pilot study, we enrolled 15 early breast cancer patients, aged 70 and older. They received a multidisciplinary CGA every 3 months and structured follow up from the SAOP nurse practitioner, dietitian, social worker, and pharmacist according to risk. Total follow-up was 6 months. Median age of evaluable patients was 79 years (range 72-87). Median number of comorbidities by Cumulative Index Rating Scale-Geriatric (CIRS-G) was 5 (3-9) at baseline. Ten patients were at pharmacological risk, five at psychosocial risk, and eight at nutritional risk. Patients presented on average six problems initially, and three new problems during follow-up. The intervention directly influenced oncological treatment in four cases. It ensured continuity/coordination of care in seven cases. Success rate in addressing problems was 87%. Mean Functional Assessment of Cancer Treatment-Breast (FACT-B) scores improved from 110.5 (S.D. 16.7) to 116.3 (S.D. 16.5) (t=0.025). Function and independence were maintained. CONCLUSIONS: Older patients with early breast cancer have a high prevalence of comorbidity. A CGA with follow-up has potential for improving the treatment and prognosis of these patients and is feasible in an academic oncology setting. PMID- 14734157 TI - Ovarian cancer in the elderly. AB - In Europe 58% of all female cancers occur in women older than 65 years. The incidence of ovarian cancer rises steadily with advancing age during adulthood and peaks in the 7th-8th decades of life. Age-specific analysis reveals that the incidence of and mortality rate from ovarian cancer are continuously increasing in the elderly population. Although more advanced stage at diagnosis seems to be one of the determinants of the worst prognosis of the elderly population, the majority of clinicians seem to be unprepared to treat elderly patients, and a great number of patients are under-treated for the fear of unacceptable side effects, thus limiting their possibility of survival. Guidelines are clearly needed; including advice on whether to treat at all and whether standard surgery or chemotherapy is feasible in elderly patients with ovarian cancer. Research on MEDLINE using as keywords 'elderly and ovarian cancer' reveals few papers, which reported data in this field. Nonetheless, in this report we will focus on four basic aspects of ovarian cancer in the elderly: the most important factors affecting prognosis, the safety of surgical treatment in aged patients, optimal first and second line chemotherapy, and the use of supportive treatments to improve quality of life. PMID- 14734158 TI - Oxalobacter formigenes and its role in oxalate metabolism in the human gut. AB - Oxalate is ingested in a wide range of animal feeds and human foods and beverages and is formed endogenously as a waste product of metabolism. Bacterial, rather than host, enzymes are required for the intestinal degradation of oxalate in man and mammals. The bacterium primarily responsible is the strict anaerobe Oxalobacter formigenes. In humans, this organism is found in the colon. O. formigenes has an obligate requirement for oxalate as a source of energy and cell carbon. In O. formigenes, the proton motive force for energy conservation is generated by the electrogenic antiport of oxalate(2-) and formate(1-) by the oxalate-formate exchanger, OxlT. The coupling of oxalate-formate exchange to the reductive decarboxylation of oxalyl CoA forms an 'indirect' proton pump. Oxalate is voided in the urine and the loss of O. formigenes may be accompanied by elevated concentrations of urinary oxalate, increasing the risk of recurrent calcium oxalate kidney stone formation. Links between the occurrence of nephrolithiasis and the presence of Oxalobacter have led to the suggestion that antibiotic therapy may contribute to the loss of this organism from the colonic microbiota. Studies in animals and human volunteers have indicated that, when administered therapeutically, O. formigenes can establish in the gut and reduce the urinary oxalate concentration following an oxalate load, hence reducing the likely incidence of calcium oxalate kidney stone formation. The findings to date suggest that anaerobic, colonic bacteria such as O. formigenes, that are able to degrade toxic compounds in the gut, may, in future, find application for therapeutic use, with substantial benefit for human health and well-being. PMID- 14734159 TI - Expression of Actinobacillus pleuropneumonia gene coding for Apx I protein in Escherichia coli. AB - This study presents cloning and expression of Actinobacillus pleuropneumoniae Apx I toxin in Escherichia coli expression system to produce fusion protein for the subsequent immunological studies. The gene coding Apx I toxin was amplified from the A. pleuropneumoniae serotype 10 DNA using polymerase chain reaction and cloned to vector under the control of strong, inducible T7 promoter. The presence of insert was confirmed by PCR screening and sequencing after the propagation of recombinant DNA in E. coli cells. The gene coding A. pleuropneumoniae Apx I toxin was extended with a segment to encode a polyhistidine tag linked to its C terminal sequence allowing a one-step affinity purification of the complex with Ni-NTA resin. Expression of the Apx I coding sequence in E. coli resulted in the formation of insoluble inclusion bodies purified according to a standard purification protocol. The ease of this expression system, the powerful single step purification and low costs make it possible to produce Apx I in large amounts to further study the role of Apx I in physiological processes. PMID- 14734160 TI - Persister cells and tolerance to antimicrobials. AB - Bacterial populations produce persister cells that neither grow nor die in the presence of microbicidal antibiotics. Persisters are largely responsible for high levels of biofilm tolerance to antimicrobials, but virtually nothing was known about their biology. Tolerance of Escherichia coli to ampicillin and ofloxacin was tested at different growth stages to gain insight into the nature of persisters. The number of persisters did not change in lag or early exponential phase, and increased dramatically in mid-exponential phase. Similar dynamics were observed with Pseudomonas aeruginosa (ofloxacin) and Staphylococcus aureus (ciprofloxacin and penicillin). This shows that production of persisters depends on growth stage. Maintaining a culture of E. coli at early exponential phase by reinoculation eliminated persisters. This suggests that persisters are not at a particular stage in the cell cycle, neither are they defective cells nor cells created in response to antibiotics. Our data indicate that persisters are specialized survivor cells. PMID- 14734161 TI - Cu/Zn superoxide dismutase in yeast mitochondria - a general phenomenon. AB - Fermentative and respiratory yeast strains of genera Saccharomyces, Kluyveromyces, Pichia, Candida and Hansenula have been investigated for mitochondrial localization of Cu/Zn superoxide dismutase (SOD). Pure mitochondrial fractions were obtained and the specific activities of Cu/Zn and Mn SODs were measured in comparison with those in the corresponding cell-free extracts. The Cu/Zn SOD: Mn SOD ratio in mitochondria and crude extracts was calculated and was considered a specific characteristic of all tested strains. Electrophoretical visualization of SOD patterns provided evidence for possible migration of cytosolic Cu/Zn SOD to mitochondria. The characteristic Cu/Zn SOD profile in mitochondria of all tested strains suggested its ubiquity within the fermentative and respiratory yeasts. PMID- 14734162 TI - Dueling quorum sensing systems in Pseudomonas aeruginosa control the production of the Pseudomonas quinolone signal (PQS). AB - The opportunistic human pathogen Pseudomonas aeruginosa regulates the production of numerous virulence factors via the action of two separate but coordinated quorum sensing systems, las and rhl. These systems control the transcription of genes in response to population density through the intercellular signals N-(3 oxododecanoyl)-L-homoserine lactone (3-oxo-C(12)-HSL) and N-(butanoyl)-L homoserine lactone (C(4)-HSL). A third P. aeruginosa signal, 2-heptyl-3-hydroxy-4 quinolone [Pseudomonas quinolone signal (PQS)], also plays a significant role in the transcription of multiple P. aeruginosa virulence genes. PQS is intertwined in the P. aeruginosa quorum sensing hierarchy with its production and bioactivity requiring the las and rhl quorum sensing systems, respectively. This report presents a preliminary transcriptional analysis of pqsA, the first gene of the recently discovered PQS biosynthetic gene cluster. We show that pqsA transcription required pqsR, a transcriptional activator protein encoded within the PQS biosynthetic gene cluster. It was also found that the transcription of pqsA and subsequent production of PQS was induced by the las quorum sensing system and repressed by the rhl quorum sensing system. In addition, PQS production was dependent on the ratio of 3-oxo-C(12)-HSL to C(4)-HSL, suggesting a regulatory balance between quorum sensing systems. These data are an important early step toward understanding the regulation of PQS synthesis and the role of PQS in P. aeruginosa intercellular signaling. PMID- 14734163 TI - Streptococcus alactolyticus is the dominating culturable lactic acid bacterium species in canine jejunum and feces of four fistulated dogs. AB - Canine intestinal lactic acid bacterium (LAB) population in four fistulated dogs was cultured and enumerated using MRS agar. LAB levels ranging from 1.4x10(6) to 1.5x10(7) CFU ml(-1) were obtained in jejunal chyme. In the fecal samples 7.0x10(7) and 2.0x10(8) CFU g(-1) were detected. Thirty randomly selected isolates growing in the highest sample dilutions were identified to species level using numerical analysis of 16S and 23S rDNA restriction fragment length polymorphism patterns (ribotyping) and 16S rDNA sequence analysis. According to these results, Streptococcus alactolyticus was the dominant culturable LAB species in both feces and jejunal chyme. In addition, Lactobacillus murinus and Lactobacillus reuteri were detected. PMID- 14734164 TI - Genetic evidence for the temperature-sensing ability of the membrane domain of the Bacillus subtilis histidine kinase DesK. AB - A decrease in environmental temperature leads to the synthesis of Delta5 unsaturated fatty acids in Bacillus subtilis by the fatty acid desaturase Des. Des is regulated by the two-component system DesKR. To understand the mechanism of cold signal perception and transduction by the membrane domain and the cytosolic domain of DesK, we expressed the cytosolic domain of DesK in trans under the control of a xylose-inducible promoter without the membrane domain. We performed growth experiments and a Northern blot analysis. Our results show that the kinase function of the cytosolic domain of DesK is temperature-independent, leading to a constitutive expression of the des gene. These findings support the conclusion that the membrane domain of DesK is the temperature-sensing element of the two-component system. PMID- 14734165 TI - Polysaccharides from the fruit bodies of the basidiomycete Laetiporus sulphureus (Bull.: Fr.) Murr. AB - The two main polysaccharides from the basidiomycetous fungus Laetiporus sulphureus were isolated, purified and characterized. The structural assignments were carried out using (13)C, (1)H, and (1)H,(13) HSQC nuclear magnetic resonance spectroscopy, methylation analysis, and Smith degradation. One was a linear beta glucan having a (1-->3)-linked main chain, namely laminaran. The other was a fucomannogalactan, which consisted of a main chain of (1-->6)-linked alpha-D galactopyranosyl residues, a part of them being substituted at O-2 by 3-O-D mannopyranosyl-L-fucopyranosyl, alpha-D-mannopyranosyl and in a minor proportion, alpha-L-fucopyranosyl groups. This heteropolysaccharide is related to those of other Basidiomycetes heterogalactans, although it differs distinctly in its side chain structures. Whereas part of the single-unit L-fucopyranosyl and/or 3-O alpha-mannopyranosyl-L-fucopyranosyl residues are present as side chains of the other heterogalactans, additional alpha-D-mannopyranosyl units are present in our fucomannogalactan of L. sulphureus. PMID- 14734166 TI - High tolerance of wild Lactobacillus plantarum and Oenococcus oeni strains to lyophilisation and stress environmental conditions of acid pH and ethanol. AB - A total of 76 Lactobacillus plantarum and Oenococcus oeni wild strains were recovered from traditionally elaborated Spanish red wines and were investigated with respect to their response to acid pH, lyophilisation, temperature and ethanol concentrations which are normally lethal to lactic acid bacteria. Both L. plantarum and O. oeni strains were able to grow at pH 3.2, were highly resistant to lyophilisation treatment and proliferated in the presence of up to 13% ethanol at 18 degrees C. Therefore, it is shown that both species are highly tolerant to stress conditions and that similarly to O. oeni strains, L. plantarum strains are of interest in beverage biotechnology. PMID- 14734167 TI - Biodiversity of cultivable psychrotrophic marine bacteria isolated from Terra Nova Bay (Ross Sea, Antarctica). AB - A set of 146 Antarctic marine isolates from the Ross Sea was characterized by a combination of molecular techniques in order to determine the degree of inter- and intraspecific variability. Isolates were analyzed by amplified rDNA restriction analysis (ARDRA) using the tetrameric enzyme AluI, resulting in 52 different groups, corresponding to at least 52 different bacterial species, indicating a high degree of interspecific variability. The phylogenetic position of bacteria belonging to some ARDRA groups was obtained by sequencing of 16S rDNA. Random amplified polymorphic DNA (RAPD) analysis, carried out on the largest ARDRA groups, revealed a high intraspecific genetic variability, too. The analysis of plasmid content revealed the existence of horizontal gene transfer between strains belonging to the same and to different species. A comparison of the whole body of morphological, physiological and biochemical data was finally carried out. PMID- 14734168 TI - PicU, a second serine protease autotransporter of uropathogenic Escherichia coli. AB - Escherichia coli is the major aetiological agent of urinary tract infections (UTI). Like diarrhoeagenic strains of E. coli, uropathogenic isolates possess virulence determinants that distinguish them from commensal strains and allow them to produce the clinical manifestations associated with UTI. Several autotransporter proteins have been associated with the ability of E. coli, and other Gram-negative bacteria, to cause disease. Recently, we described the existence within uropathogenic E. coli (UPEC) strains of Sat, a toxin of the serine protease autotransporter of Enterobacteriaceae (SPATE) subfamily. Using features common to proteins secreted via the autotransporter pathway we have identified nine additional autotransporter proteins from the genomic sequence data of UPEC CFT073. Surprisingly, two additional members of the SPATE subfamily were identified. One protein, designated PicU, was homologous to the Pic protein identified in Shigella flexneri and enteroaggregative E. coli. The PicU protein was expressed and investigated for functional activity. PMID- 14734169 TI - Cloning, sequencing and functional analysis of Magnaporthe grisea MVP1 gene, a hex-1 homolog encoding a putative 'woronin body' protein. AB - A hex-1 homolog named MVP1 was isolated from an appressoria cDNA library of the rice blast fungus Magnaporthe grisea. The transcript of approximately 1.6 kb contains 546 bp of coding sequence with a 3' untranslated region about 168 bp long and a 5' untranslated region about 870 bp long. Southern gel blot analysis of genomic DNA following digestion with three restriction enzymes (BamHI, EcoRI, HindIII) indicated that the gene exists as a single copy in M. grisea genome. RNA gel blot analyses showed that MVP1 was highly expressed in germinating conidia and the mycelial stage compared to appressoria or non-germinated conidia. MVP1 showed a high degree of homology to the hex-1 gene recently described to encode a major protein in the woronin bodies of Neurospora crassa. Double homologous recombination was used to replace MVP1 with the hyg(R) gene. MVP1 knockout mutants showed apical swellings when grown on agar plates containing 2% sorbose but they were not impaired in any other vegetative or pathogenic properties evaluated. The pathological and other phenotypic consequences of gene disruption are discussed. PMID- 14734170 TI - Expression of the landomycin biosynthetic gene cluster in a PKS mutant of Streptomyces fradiae is dependent on the coexpression of a putative transcriptional activator gene. AB - The formation of landomycin A or one of its derivatives (5,6-anhydrolandomycin A) in a heterologous strain has never been achieved. It has now been made possible by the coexpression of a cosmid containing all biosynthetic genes necessary to produce landomycin A together with a pathway-specific regulatory gene. As host we used a polyketide synthase-defective mutant strain of Streptomyces fradiae Tu2717 which is not able to produce urdamycin A. Our results indicate that four glycosyltransferases are responsible for the formation of the hexasaccharide side chain of landomycin A. PMID- 14734171 TI - The ATPase domain of HscC (DnaK homolog) is essential for interfering sigma70 activity in E. coli. AB - HscC, a DnaK homolog in Escherichia coli, consists of adenosine triphosphatase (ATPase), substrate-binding and C-terminal domains. Overexpression of HscC markedly inhibits growth of host cell and reduces the sigma(70)-dependent promoter activity presumably by forming a complex with sigma(70). To identify the region(s) of HscC responsible for growth inhibition and complex formation with sigma(70), domain swapping experiments were carried out between DnaK and HscC. Thus the chimeric proteins carrying the ATPase domain of HscC and substrate binding domains of either HscC or DnaK were found to inhibit the growth of the cell, reduce the sigma(70)-dependent promoter activity and form a complex with sigma(70). These results indicate that the ATPase domain of HscC rather than the substrate-binding domain is important for determining its functional specificity. PMID- 14734172 TI - Role of exopolysaccharide, the rugose phenotype and VpsR in the pathogenesis of epidemic Vibrio cholerae. AB - Vibrio cholerae, the causative agent of cholera can produce an exopolysaccharide (EPS). Some strains can also phenotypically switch from a smooth to a 'rugose' phenotype characterized by small wrinkled colonies, overproduction of EPS, increased biofilm formation in vitro and increased resistance to various stressful conditions. High frequency switching to the rugose phenotype is more common in epidemic strains than in non-pathogenic strains, suggesting EPS production and the rugose phenotype are important in cholera epidemiology. VpsR up-regulates Vibrio polysaccharide (VPS) genes and the synthesis of extracellular EPS (VPS). However, the function of VPS, the rugose phenotype and VpsR in pathogenesis is not well understood. We report that rugose strains of both classical and El Tor biotypes of epidemic V. cholerae are defective in the in vitro production of extracellular collagenase activity. In vivo studies in rabbit ileal loops suggest that VpsR mutants are attenuated in reactogenicity. Intestinal colonization studies in infant mice suggest that VPS production, the rugose phenotype and VpsR have a role in pathogenesis. Our results indicate that regulated VPS production is important for promoting in vivo biofilm formation and pathogenesis. Additionally, VpsR might regulate genes with roles in virulence. Rugose strains appear to be a subpopulation of cells that might act as a 'helper' phenotype promoting the pathogenesis of certain strains. Our studies provide new insight into the potential role of VPS, the rugose phenotype and VpsR in the pathogenesis of epidemic V. cholerae. PMID- 14734173 TI - Ornithine and arginine decarboxylase activities and effect of some polyamine biosynthesis inhibitors on Gigaspora rosea germinating spores. AB - The pathways for putrescine biosynthesis and the effects of polyamine biosynthesis inhibitors on the germination and hyphal development of Gigaspora rosea spores were investigated. Incubation of spores with different radioactive substrates demonstrated that both arginine and ornithine decarboxylase pathways participate in putrescine biosynthesis in G. rosea. Spermidine and spermine were the most abundant polyamines in this fungus. The putrescine biosynthesis inhibitors alpha-difluoromethylarginine and alpha-difluoromethylornithine, as well as the spermidine synthase inhibitor cyclohexylamine, slightly decreased polyamine levels. However, only the latter interfered with spore germination. The consequences of the use of putrescine biosynthesis inhibitors for the control of plant pathogenic fungi on the viability of G. rosea spores in soil are discussed. PMID- 14734174 TI - Changes in properties of glutamate transport in Trichoderma viride vegetative mycelia upon adaptation to glutamate as carbon source. AB - The U-(14)C-labelled glutamate uptake was measured in both sucrose- and glutamate grown mycelia of Trichoderma viride. The biomass yield was five-fold lower with glutamate as a sole carbon source. The rate of glutamate transport measured at a glutamate concentration of 1 mM remained unchanged in glutamate-grown mycelia whereas the properties of the glutamate transport were substantially changed compared to sucrose-grown mycelia. The glutamate uptake in both sucrose- and glutamate-grown mycelia was inhibited by an uncoupler (3,3',4',5 tetrachlorosalicylanilide) but the inhibitory efficiency was higher in the latter. The affinity of the permease to glutamate increased approximately five fold in the glutamate-grown mycelia (about 76 microM compared to about 16 microM). The pH optimum for glutamate uptake was 4 in sucrose-grown mycelia but the glutamate-grown mycelia had two pH optima, one at pH 4 and the second between pH 6 and 7. The inhibition of glutamate uptake by other amino acids yielded different inhibitory patterns in the two mycelia under study. The glutamate uptake in mycelia of different ages also showed differences in both transport rate and temporal pattern. The results show that the growth of mycelia on glutamate led to the appearance of an additional permease with different properties and suggest that only this permease is operating in mycelia grown on glutamate. PMID- 14734175 TI - Heterologous production and characterization of bacterial nickel/cobalt permeases. AB - Nickel/cobalt permeases (NiCoTs, TC 2.A.52) are a rapidly growing family of structurally related membrane transporters whose members are found in Gram negative and Gram-positive bacteria, in thermoacidophilic archaea, and in fungi. Previous studies have predicted two subclasses represented by HoxN of Ralstonia eutropha, a selective nickel transporter, and by NhlF of Rhodococcus rhodochrous, a nickel and cobalt transporter that displays a preference for the Co ion. In the present study, NiCoT genes of five Gram-negative bacteria and one Gram-positive bacterium were cloned and heterologously expressed in Escherichia coli. Based on substrate preference in metal-accumulation assays with the recombinant strains, two of the novel NiCoTs were assigned to the NhlF class. The remaining four NiCoTs belong to a yet unrecognized, third class. They transport both the nickel and the cobalt ion but have a significantly higher capacity for nickel. The observed substrate preferences correlate in many cases with the genomic localization of NiCoT genes adjacent to regions encoding nickel- or cobalt dependent enzymes or enzymes involved in cobalamin biosynthesis. Alignment of 23 full-length NiCoT sequences and comparison with the available experimental data predict that substrate specificity of NiCoTs is an adaptation to specific transition metal requirements in various organisms from different taxa. PMID- 14734176 TI - Cloning and characterization of preferentially expressed genes in an aluminum tolerant mutant derived from Penicillium chrysogenum IFO4626. AB - cDNAs expressed preferentially in an Al-tolerant microorganism were isolated by subtraction hybridization with cDNAs of Al-sensitive Penicillium chrysogenum IFO4626 as driver cDNA and cDNAs of the Al-tolerant mutant derived from the wild cells by UV irradiation as tester cDNA. Northern blot analysis revealed that mRNA levels of six genes were increased significantly in the Al-tolerant mutant after exposure to Al stress when compared with the wild cells. Two genes accumulated in both the presence and absence of Al stress and four genes were induced by Al stress in the Al-tolerant mutant. cDNA fragments were amplified by rapid amplification of cDNA ends and sequenced to obtain full-length cDNAs of the six genes. Two genes were novel or predicted ones and the others showed significant homology to known genes, ADP/ATP translocase, enolase, cysteine synthase, and glucoamylase, which are induced by environmental stresses in prokaryotic and eukaryotic cells. These enzyme activities increased in the Al-tolerant mutant when compared to those in the wild cells, showing that not only the levels of gene expression but also the levels of enzyme activities increased in the Al tolerant mutant. PMID- 14734177 TI - Detection of an IS21 insertion sequence in the mexR gene of Pseudomonas aeruginosa increasing beta-lactam resistance. AB - To understand the regulation of the MexAB OprM efflux system in a clinical strain of Pseudomonas aeruginosa presenting a decreased susceptibility to ticarcillin and aztreonam, the mexR repressor gene was amplified by polymerase chain reaction (PCR) and was shown to be disrupted by an insertion sequence of more than 2 kb, with characteristic direct and inverted repeat sequences. Sequencing revealed a 2131-bp IS21 insertion sequence. A reverse transcription PCR method was used to quantify mexA transcripts and showed an increased transcription rate of mexA in this strain, compared with a PAO1 control strain. The nalB phenotype in P. aeruginosa may be due to point mutations, but also to the presence of an insertion sequence in the mexR regulator gene. PMID- 14734178 TI - Infection of murine macrophage cell lines by Legionella pneumophila. AB - Legionella pneumophila causes pneumonia by infecting alveolar macrophages. Although several model systems have been used for L. pneumophila virulence studies, no detailed comparisons have been made between them. An ideal in vitro virulence model should be cost-effective, easy to obtain in large amounts and as relevant as possible to the actual disease. We compared the MH-S cell line to human peripheral blood monocyte-derived macrophages and the J774A.1 cell line. We found that the interactions of L. pneumophila with MH-S at the cellular level resemble those of human primary monocyte-derived macrophages, suggesting that these cells provide a valuable model for this bacterial pathogen. PMID- 14734179 TI - Genetic variation of Cordyceps sinensis, a fruit-body-producing entomopathogenic species from different geographical regions in China. AB - Cordyceps sinensis is one of the most valuable medicinal fungi in the Orient. It is naturally distributed in the eastern extension area of the Qinghai-Tibet plateau, at an altitude over 4000 m high. In order to investigate genetic variation and evolutionary relationships of C. sinensis from different geographical regions, 17 isolates of C. sinensis were collected from different provinces and the complete sequences of rDNA ITS were determined. On the basis of 5.8S rDNA and ITS region analysis, it was clearly shown that the ITS sequences within C. sinensis are highly homologous regardless of geographical origin. The distance values between the sequences in this study were lower than 0.03. This implied that C. sinensis from different geographic regions are the same species; they are not different species or a species complex. The results also showed that distance values between C. sinensis and Hirsutella sinensis are of the same order as those within C. sinensis from different geographic regions. This confirmed our previous results that C. sinensis should only have H. sinensis as its asexual stage whatever the geographic region from which the samples were collected. An rDNA ITS clone library was established to obtain further evidence for the interpretation of the fungal community structure from C. sinensis and to confirm the accuracy of the taxonomic identities produced by directly sequencing the rDNA ITS region. The discrimination between intraspecies of C. sinensis might provide additional data for the authentication of medicinal material at the species or intraspecies level. PMID- 14734180 TI - Regulation of virulence determinants in vitro and in vivo in Staphylococcus aureus. AB - Staphylococcus aureus is an opportunistic pathogen. In response to changing host environments, this bacterium has the capability to switch on selective sets of genes to enhance its chances for survival. This switching process is precisely controlled by global regulatory elements. There are two major groups of global regulatory elements in S. aureus, including two-component regulatory systems (TCRSs) and the SarA protein family. Presumably, the sensor proteins of the 16 TCRSs in S. aureus provide external sensing, while the response regulators, in conjunction with alternative transcription factors and the SarA protein family, function as effectors within the intricate regulatory network to respond to environmental stimuli. Sequence alignment and structural data indicate that the SarA protein family could be subdivided into three subfamilies: (1) single-domain proteins; (2) double-domain proteins; and (3) proteins homologous to the MarR protein family. Recent data using reporter gene fusions in animal models, have confirmed distinct expression profiles of selected regulatory and target genes in vitro vs. in vivo. PMID- 14734181 TI - Simple sequence repeats (microsatellites): mutational mechanisms and contributions to bacterial pathogenesis. A meeting review. AB - This review summarises the presentations and discussions that took place during a European Science Foundation-funded workshop whose purpose was to gain current perspectives on the mutational mechanisms of simple sequence repeats and the contribution of localised hypermutation in such repeats to bacterial pathogenesis. In vitro biophysical and biochemical assays of mutational mechanisms were covered as well as genetic studies in various eukaryotic and prokaryotic organisms. Presentations on bacterial pathogenesis elaborated investigations of the use of repeats for typing of strains, epidemiological investigations of mutation rates and functions of loci whose expression is controlled by simple sequence repeats. This review tabulates current perspectives on the cis- and trans-acting factors for mutation of simple sequence repeats and the orientations of mononucleotide repeats in some bacterial species that utilise repeats for adaptation. PMID- 14734182 TI - Bacteriophage-mediated nucleic acid immunisation. AB - Whole bacteriophage lambda particles, containing reporter genes under the control of the cytomegalovirus promoter (P(CMV)), have been used as delivery vehicles for nucleic acid immunisation. Following intramuscular injection of mice with lambda gt11 containing the gene for hepatitis B surface antigen (HBsAg), anti-HBsAg responses in excess of 150 mIU ml(-1) were detected. When isolated peritoneal macrophages were incubated with whole lambda particles containing the gene for green fluorescent protein (GFP) under the control of P(CMV), GFP antigen was detected on the macrophage surface 8 h later. Results suggested that direct targeting of antigen-presenting cells by bacteriophage 'vaccines' may occur, leading to enhanced immune responses compared to naked DNA delivery. Bacteriophage DNA vaccines offer several advantages: they do not contain antibiotic resistance genes, they offer a large cloning capacity (approximately 15 kb), the DNA is protected from environmental degradation, they offer the potential for oral delivery, and large-scale production is cheap, easy and extremely rapid. PMID- 14734183 TI - Generation of Marburg virus-like particles by co-expression of glycoprotein and matrix protein. AB - Marburg virus (MARV), the causative agent of a severe hemorrhagic fever, has a characteristic filamentous morphology. Here we report that co-expression of MARV glycoprotein and matrix protein (VP40) in mammalian cells leads to spontaneous budding of filamentous particles strikingly similar to wild-type MARV. In addition, these particles elicit an immune response in BALB/c mice. The generation of non-replicating Marburg virus-like particles (VLPs) should significantly facilitate the research on molecular mechanisms of MARV assembly and release. Furthermore, VLPs may be an excellent vaccine candidate against Marburg infection. PMID- 14734184 TI - Comparative analysis of amplified fragment length polymorphism and pulsed field gel electrophoresis in a hospital outbreak and subsequent endemicity of ampicillin-resistant Enterococcus faecium. AB - Reliable molecular methods for determination of relatedness between bacterial isolates have become increasingly important to evaluate outbreaks and endemic situations with nosocomial pathogens. In the present study Simpson's index of diversity with calculated confidence intervals was used to compare amplified fragment length polymorphism (AFLP) and pulsed field gel electrophoresis (PFGE) analysis of a hospital outbreak of ampicillin-resistant Enterococcus faecium and subsequent endemicity. The outbreak, in a Norwegian tertiary hospital, of infections caused by these enterococci started in 1995 and increased in 1996 after which the situation turned endemic. The purpose of this study was to compare the two methods in this setting and to determine the length of time during an outbreak that these methods are sufficiently valid to be of value for hospital infection control efforts. One hundred and sixty clinical isolates from urine specimens collected during the period 1995-1999 were included. The findings indicate that PFGE and AFLP are equally discriminative and could in this setting be used for typing purposes over the whole 5-year period. PMID- 14734185 TI - Identification of pathogenic dematiaceous fungi and related taxa based on large subunit ribosomal DNA D1/D2 domain sequence analysis. AB - The nucleotide sequences of the D1/D2 domains of large subunit (26S) ribosomal DNA for 76 strains of 46 species of pathogenic dematiaceous fungi and related taxa were determined. Intra-species sequence diversity of medically important dematiaceous fungi including Phialophora verrucosa, Fonsecaea pedrosoi, Fonsecaea compacta, Cladophialophora carrionii, Cladophialophora bantiana, Exophiala dermatitidis, Exophiala jeanselmei, Exophiala spinifera, Exophiala moniliae, and Hortaea werneckii were extremely small; as few as 0 changes were detected in C. bantiana, Fonsecaea and Exophiala species, 1 bp in C. carrionii and H. werneckii, and 2 bp in P. verrucosa. Inter-species nucleotide diversity between most species was higher. These data suggested that the D1/D2 domain is sufficiently variable for identification of pathogenic dematiaceous fungi and relevant species. The phylogenetic trees constructed from the sequence data revealed that most human pathogenic species formed a single cluster and that Cladosporium and Phialophora species were distributed polyphyletically into several clusters. PMID- 14734186 TI - Yersiniabactin and other siderophores produced by clinical isolates of Enterobacter spp. and Citrobacter spp. AB - We analyzed the ability of extraintestinal strains of Enterobacter spp. and Citrobacter spp. to employ different siderophore-mediated strategies of iron acquisition. All strains produced iron-chelating compounds. Cross-feeding assays indicated that most isolates of both Enterobacter spp. and Citrobacter spp. excreted catecholate siderophore enterobactin, less produced aerobactin, and single strains excreted hydroxamates different from aerobactin. Besides, we analyzed if the strains had the ability to produce the siderophore yersiniabactin coded by the Yersinia high-pathogenicity island (HPI). The presence of HPI genes was observed in single isolates of three species: E. cloaceae, E. aerogenes and C. koseri. A detailed polymerase chain reaction analysis revealed differences in the genetic organization of the HPIs; however, in a cross-feeding test we proved that yersiniabactin was produced and the island was functional. PMID- 14734187 TI - Detection of high-level tetracycline resistance in clinical isolates of Helicobacter pylori using PCR-RFLP. AB - Tetracycline is one of four antibiotics commonly used for the treatment of Helicobacter pylori infection, but its effectiveness is decreasing as the incidence of tetracycline resistance is increasing. In five Brazilian tetracycline-resistant (Tet(R)) H. pylori isolates, high-level tetracycline resistance is mediated by the triple-base-pair substitution AGA(926-928)-->TTC in both 16S rRNA genes, as was previously observed in two independent high-level Tet(R) H. pylori strains. A polymerase chain reaction-based restriction fragment length polymorphism (PCR-RFLP) assay was developed for the detection of the AGA(926-928)-->TTC substitution, and confirmed the presence of the aforementioned triple-base-pair substitution in all five Brazilian Tet(R) isolates. This PCR RFLP-based approach distinguishes the high-level Tet(R) isolates from low-level Tet(R) and Tet(S) H. pylori strains and thus allows the direct detection of Tet(R) H. pylori isolates. PMID- 14734188 TI - Disruption of CD40/CD40L interaction influences the course of Cryptococcus neoformans infection. AB - CD40 signaling has been implicated in various pathogenic processes such as chronic inflammatory disease, graft-versus-host disease, autoimmune disease and cancer. We previously demonstrated in an in vitro system that the CD40/CD40L pathway mediates late interleukin (IL) 12 production in response to Cryptococcus neoformans. The purpose of this study was to examine the course of C. neoformans infection in the absence of CD40/CD40L costimulation. We compared infection in mice genetically lacking CD40L (CD40L(-/-)) and in the wild-type counterpart. The animals were injected intratracheally with C. neoformans and monitored for clearance of the organism and the development of cellular immune response. CD40L( /-) mice exhibited an exacerbation of infection, evaluated as scarce inflammatory response in the lung, that mirrored an increase of fungal burden. This correlated with impairment of nitrite production and antimicrobial activity by macrophages against C. neoformans and unrelated microorganisms such as Candida albicans. Moreover, IL-12 production by splenic macrophages was diminished in CD40L(-/-) mice and interferon-gamma production by CD4 and CD8 T cells was decreased. CD4 T cells retained the ability to express a costimulatory molecule, CTLA-4, but showed a decrease in CD28 expression. This latter molecule is implicated in a positive effect on proliferation, cytokine production and survival of T cells. Collectively these data demonstrate that absence of CD40L correlates with (i) reduced antimicrobial activity of natural effector cells; (ii) reduction of the magnitude of T cell response; and (iii) increase of fungal growth in the brain. These findings suggest that disruption of CD40/CD40L may be deleterious for development of an efficient immune response to C. neoformans and may identify potential molecular targets for novel immunotherapeutic approaches PMID- 14734189 TI - Treatment of anthrax infection with combination of ciprofloxacin and antibodies to protective antigen of Bacillus anthracis. AB - Currently there is no effective treatment for inhalational anthrax beyond administration of antibiotics shortly after exposure. There is need for new, safe and effective treatments to supplement traditional antibiotic therapy. Our study was based on the premise that simultaneous inhibition of lethal toxin action with antibodies and blocking of bacterial growth by antibiotics will be beneficial for the treatment of anthrax. In this study, we tested the effects of a combination treatment using purified rabbit or sheep anti-protective antigen (PA) antibodies and the antibiotic ciprofloxacin in a rodent anthrax model. In mice infected with a dose of Bacillus anthracis Sterne strain corresponding to 10 LD(50), antibiotic treatment with ciprofloxacin alone only cured 50% of infected animals. Administration of anti-PA IgG in combination with ciprofloxacin produced 90-100% survival. These data indicate that a combination of antibiotic/immunoglobulin therapy is more effective than antibiotic treatment alone in a rodent anthrax model. PMID- 14734190 TI - Evaluation of immunogenicity and protective efficacy of a Microsporum canis metalloprotease subunit vaccine in guinea pigs. AB - In order to identify protective immunogens against Microsporum canis infection, a purified recombinant keratinolytic metalloprotease (r-MEP3) was tested as a subunit vaccine in experimentally infected guinea pigs. Both humoral and cellular specific immune responses developing towards r-MEP3 were evaluated, by enzyme linked immunosorbent assay and by in vitro lymphocyte transformation tests respectively. Vaccination induced a strong antibody response, and a significant but transient lymphoproliferative response against the protein. However, the protocol failed to prevent fungal invasion or development of dermatophytic lesions. These results show that under the present experimental conditions, r MEP3 specific antibodies are not protective against a challenge exposure. They also suggest that in the same model, the induction of cell-mediated immunity towards r-MEP3 is not sufficient, indicating the need for further research in the field of specific immune mechanisms involved in M. canis dermatophytosis. PMID- 14734191 TI - The diversity of vacA and cagA genes of Helicobacter pylori in East Asia. AB - It has been reported that Helicobacter pylori infection with the type I strain, which expresses the VacA and CagA antigens, is associated with duodenal ulcer. We examined the diversity of vacA and cagA genes in 143 isolates obtained from patients with duodenal ulcer or chronic gastritis in East Asia (two different areas of Japan, Fukui and Okinawa, and also in Hangzhou, China) by polymerase chain reaction (PCR) and sequence analysis. Diversities of cagA and vacA genes were detected in East Asia. The prevalence of cagA-positive H. pylori was significantly different between Fukui and Okinawa (P=0.0032). The prevalence of Western type CagA was significantly higher in Okinawa than in Fukui (P<0.0001). However, there was no significant association between the genotype of cagA and clinical outcome. In Japan, the predominant vacA genotype was s1c/m1b. In contrast, in Hangzhou, the predominant vacA genotype was s1c/m2, and they were all East Asian CagA-positive. These findings suggest that a distinct distribution of the vacA and cagA genotypes is present in East Asia, regardless of clinical outcome. PMID- 14734192 TI - Analysis of Moraxella catarrhalis outer membrane antigens cross-reactive with Neisseria meningitidis and Neisseria lactamica. AB - Mouse sera against outer membrane proteins from Moraxella catarrhalis, Neisseria meningitidis and Neisseria lactamica, and human sera from both healthy individuals and patients convalescing from meningococcal meningitis were used to identify cross-reactive antigens. Mouse anti-N. meningitidis and anti-N. lactamica sera recognized 77, 62 and 32 kDa outer membrane antigens in M. catarrhalis strains; on the contrary, the meningococcal porin PorB (38-42 kDa) was recognized by one of the two anti-M. catarrhalis sera. Human sera from both healthy individuals and patients convalescing from meningococcal meningitis also showed cross-reactive antibodies against these proteins. The existence of cross reactive antigens in M. catarrhalis and N. meningitidis (as well as in N. lactamica) could favor the development of natural immunization against both pathogens. PMID- 14734193 TI - The clinical value of intracellular autoantigens B-cell epitopes in systemic rheumatic diseases. AB - A hallmark of autoimmune diseases is the production of autoantibodies against intracellular autoantigens. Although their pathogenetic and their etiologic relationship are not fully understood, these autoantibodies are important tools for establishing the diagnosis, classification and prognosis of autoimmune diseases. Systemic rheumatic diseases are among the most complex disorders because their clinical presentation and constellation of findings are in part reflected by the wide spectrum of autoantibodies found in the sera of patients suffering from these disorders. These autoantibodies usually target large complexes consisting of protein antigens noncovalently associated with (ribo) nucleic acid(s), like the spliceosome or Ro/La-RNPs. In this review, we first address the main characteristics and the clinical value of several autoantibodies, with respect to their diagnostic sensitivity and specificity. Subsequently, we provide a brief overview of the antigenic determinant types that have been identified on the corresponding autoantigens. The antibody targets of autontigens include primary, secondary, tertiary and quarternary structure epitopes, as well as cryptotopes, neoepitopes and mimotopes. We next focus on antigenic structures corresponding to B-cell epitopes with high disease specificity and sensitivity for all the major autoantigens in systemic autoimmunity including the Ro/La and U1 ribonucleoprotein complexes and the Ku70/80, ribosomal P, DNA topoisomerase I, filaggrin, Jo-1 and PM/SCl-100 autoantigens. These epitopes, defined at the peptide level, can be chemically synthesized and engineered for the development of new inexpensive and easier to perform assays and the improvement of the methods for autoantibody detection. Specific examples of newly developed assays that incorporate (i) epitopes with high disease specificity and sensitivity, (ii) modified epitopes, (iii) conformational epitopes and (iv) complementary epitopes are discussed in detail. Finally, we examine the potential of combining these synthetic epitopes for future development of multiplex diagnostic tests based on miniaturized autoantigen arrays. PMID- 14734194 TI - Role of the ubiquitin-proteasome pathway in the diagnosis of human diseases. AB - The ubiquitin-proteasome pathway constitutes the major system for nuclear and extralysosomal cytosolic protein degradation in eukaryotic cells. A plethora of cell proteins implicated in the maintenance and regulation of essential cellular processes undergoes processing and functional modification by proteolytic degradation via the ubiquitin-proteasome pathway. Deregulations of the pathway have been shown to contribute to the pathogenesis of several human diseases, such as cancer, neurodegenerative, autoimmune, genetic and metabolic disorders, most of them exhibiting abnormal accumulation and altered composition of components of the pathway that is suitable for diagnostic proceedings. While the ubiquitin proteasome pathway is currently exploited to develop novel therapeutic strategies, it is less regarded as a diagnostic area. Future research should lead to an improved understanding of the pathophysiology of the ubiquitin-proteasome pathway with the aim of allowing the development of subtle diagnostic strategies. PMID- 14734195 TI - Laboratory testing for endometriosis. AB - BACKGROUND: Typically, endometriosis is diagnosed surgically by laparoscopy. CA 125 is the principal serum marker used in the diagnosis and management of late stage endometriosis. The search for a body fluid marker of early stage disease has included studies of serum, peritoneal fluid (PF), and/or tissue levels of secretory proteins, cell adhesion molecules, cytokines, tumor necrosis and vascular endothelial growth factors (VEGFs), chemokines, antiendometrial antibodies, autoantibodies to oxidized lipoproteins, aromatase P-450 expression, cytokeratins, and hormone receptors. We compared the diagnostic accuracy and clinical utility of these various types of substances in the non-surgical identification of patients with endometriosis. METHOD: We reviewed the MEDLINE database for all publications on serum, peritoneal fluid and tissue markers of endometriosis. RESULTS: Except for serum interleukin (IL)-6 and peritoneal fluid tumor necrosis factor (TNF)-alpha levels, the diagnostic accuracy of other markers of endometriosis was either similar or worse than that of CA-125 (sensitivity 24-94%; specificity 83-93%). The diagnostic accuracy of IL-6 and TNF alpha was 90-100% (sensitivity) and 67-89% (specificity). CONCLUSION: CA-125 has limited diagnostic accuracy in the identification of early stage endometriosis and none of the other markers we reviewed dramatically outperformed CA-125 in this regard with the possible exception of serum IL-6 and peritoneal fluid TNF alpha levels. PMID- 14734196 TI - Urinary markers of bladder carcinoma. AB - PURPOSE: Transitional cell carcinoma of bladder is one of the most common tumors of genitourinary tract. Cystoscopy along with cytology is the mainstay for diagnosing bladder cancer. Cytology is specific for diagnosis of bladder carcinomas but less sensitive particularly in low-grade disease. Cystoscopy on the other hand is invasive and relatively costly technique and may also be inconclusive at times particularly in case of cystitis. A simple noninvasive marker for detecting bladder cancer would be helpful. A clinically useful urinary marker should be easy to perform, should have minimum requirements for sample preparations and should have high sensitivity and specificity in diagnosis. In this review we discussed the various urinary markers and their role in detection of bladder cancer. MATERIALS AND METHODS: We reviewed the literature on urinary markers and tests that may have clinical usefulness. RESULTS: Several urinary markers and tests such as BTA Stat, BTA TRAK, NMP22, telomerase, HA and HAse tests, Immunocyt, Quanticyt, FDP, BLCA-4, FISH, CYFRA-21-1 have enough potential for future clinical use. BTA stat, NMP22 (bladder check)and AccuDX (FDP) tests are presently point of care tests. The rest of the tests are laboratory-based and may need trained technicians. Majorities of the urinary markers have higher sensitivity and specificity than cytology. However, voided urinary cytology has the highest specificity. CONCLUSIONS: Till now there is no urinary marker or test that can replace the need of cystoscopy. However, some markers have the potential for future clinical use. PMID- 14734197 TI - Advances in the clinical laboratory assessment of urinary sediment. AB - Urinalysis has been used extensively in clinical practice to aid in the diagnosis of various renal and urologic diseases. The innovation of urinalysis is marching on right along with the rapid developments in biotechnology and astride from the solo urine cytology to sophisticated studies of individual component in the urinary sediment. In this review article, we focus on the use of flow cytometry and other technical advances in the examination of urinary sediment, the detection of urologic malignancies by the presence of microsatellite alteration in the urinary sediment, as well as the quantification of cytokine messenger RNA (mRNA) expression in urinary sediment by reverse transcription and real-time quantitative polymerase chain reaction (RT-QPCR). Notably, the study of cytokine mRNA expression in urinary sediment by RT-QPCR has recently been reported to provide important diagnostic information in kidney allograft recipients and patients with lupus nephritis. This simple and non-invasive method requires further study to determine its role in risk stratification and monitoring of therapeutic response in patients with other kidney diseases. PMID- 14734198 TI - Relationship between malondialdehyde level and glutathione peroxidase activity in diabetic rats. AB - BACKGROUND: This investigation describes the relationship between glutathione peroxidase activities, an antioxidant enzyme, and the oxidative status in diabetic rat blood. METHODS: Malondialdehyde level and glutathione peroxidase activity were measured by spectrophotometric techniques. RESULTS AND CONCLUSIONS: Malondialdehyde content in the diabetic rats group was increased compared to that in the controls [3.08+/-0.32 (mean+/-S.E.) vs. 1.15+/-0.29 mM/g hemoglobin, P>0.01]. Glutathione peroxidase activity in the diabetic rats group was increased compared to that in the control [10.27+/-1.39 (mean+/-S.E.) vs. 3.14+/-0.38 micromol NADPH/min/g hemoglobin, P>0.01]. Our results show a positive correlation between serum glutathione peroxidase and malondialdehyde concentration in diabetic rats. PMID- 14734199 TI - Evaluation of two-dimensional cytometric lamellar body counts on the ADVIA 120 hematology system for estimation of fetal lung maturation. AB - BACKGROUND: We sought to determine if the ADVIA 120 hematology system (Bayer HealthCare LLC, Diagnostics Division, Tarrytown, NY), which employs a unique two dimensional cytometric analysis approach for platelet (PLT) counting, might also be useful for estimation of LB lamellar body counts in amniotic fluid. METHODS: ADVIA 120 LB counts were performed on 217 amniotic fluid specimens, 88 of which were obtained within 72 h of infant delivery. The ADVIA 120 LB count (ADVIA LB count) results were compared with results from other FLM tests, including the TDx FLM II (FLM II) test (Abbott Diagnostics, Abbott Park, IL) and the A(650) estimate of lamellar body density using the refractive-index-matched anomalous diffraction (RIMAD) technique. RESULTS: Use of an ADVIA LB count referent value of > or =35,400/microl to indicate maturity, yielded sensitivity, specificity, PV(RDS) and PV(maturity) of 100%, 67.6%, 36.8% and 100%, respectively, in our study population (prevalence of RDS=15.9%). The clinical performance of the ADVIA LB count assay was compared to that of the FLM II and RIMAD assays by predictive value and receiver operating characteristic (ROC) curve analysis. CONCLUSIONS: These data suggest that the ADVIA 120-derived LB counts on amniotic fluid can be useful in predicting fetal lung maturity. PMID- 14734200 TI - Increased electronegative charge of serum low-density lipoprotein in patients with diabetes mellitus. AB - BACKGROUND: Patients with diabetes mellitus have been reported to show increased serum levels of modified low-density lipoprotein (LDL), including glycosylated, oxidized and small, dense LDL. This change has been suggested to represent an important risk factor for diabetic macroangiopathy. A common characteristic shared by these modified LDL species is the increase in electronegative charge on particle surfaces, which can be detected by agarose gel electrophoresis as "LDL charge modified frequency" (LDL-CMF) determined from the relative mobility of LDL fraction. METHODS: LDL-CMF was measured in the sera from 129 outpatients with type 2 diabetes mellitus and compared with the data from 34 normal subjects. RESULTS: The LDL fraction from diabetics migrates more closely to the anode side as compared with that from normal subjects. The LDL-CMF measured in diabetics, 5.5+/-8.1%, was significantly (p<0.0001) higher than 0.6+/-3.4% in normal subjects. Serum LDL-CMF showed significant positive correlations with triglyceride at r=0.552 (p<0.0001) and malondialdehyde modified LDL at r=0.390 (p<0.0001), as well as systolic blood pressure, body mass index, fasting plasma glucose, hemoglobin A(1c), total cholesterol, free fatty acid (FFA) and homeostasis model assessment ratio. It showed negative correlations with high density lipoprotein and total superoxide dismutase activity. CONCLUSION: The results indicate that LDL-CMF reflects the degree of serum LDL modification in diabetics and can be regarded as an important risk factor for diabetic macroangiopathy. PMID- 14734201 TI - Methylenetetrahydrofolate reductase (MTHFR) c677t gene variant modulates the homocysteine folate correlation in a mild folate-deficient population. AB - BACKGROUND: A large body of evidence links plasma concentrations of homocysteine and cardiovascular disease. Several genetic and environmental variables may modulate such relationship. We investigated the influence of methylenetetrahydrofolate reductase (MTHFR) gene variants C677T, A1298C, and T1317C on homocysteine, folate, and cobalamin concentrations in a sample of individuals from a mild folate deficiency population to better clarify the complex interactions existing among these variables. METHODS: In the present study, 209 individuals belonging to an admixed urban population characterized by mild folate deficiency were investigated. MTHFR gene variants C677T, A1298C, and T1317C were genotyped and homocysteine-, folate-, and cobalamin-determined for each individual. RESULTS: Univariate analyses showed a significant association between the C677T variant with homocysteine (P<0.001) and cobalamin (P=0.005) as well as a significant relationship between the T allele and serum folate concentrations (P<0.05). The TT genotype of the C677T polymorphism remained significantly associated with log-transformed homocysteine even after adjustment for age, sex, smoking status, ethnicity, folate, and cobalamin concentrations (P<0.01). Both univariate and multivariate analysis have failed to show any effect of the A1298C and T1317C genetic variants in homocysteine concentrations in this population. Finally, a significant interaction between folate and C677T polymorphism in the determination of homocysteine was also disclosed (P<0.005). CONCLUSIONS: Taken together, these results demonstrate a significant interaction between serum folate and MTHFR genotype in predicting homocysteine concentrations. One may consider that a differential response of homocysteine to folic acid supplementation may depend on MTHFR genotype which may have important implications when attempting to lower homocysteine concentrations in populations with mild folate deficiency. PMID- 14734202 TI - Effects of vitamin E on oxidative stress and membrane fluidity in brain of streptozotocin-induced diabetic rats. AB - BACKGROUND: Diabetics and experimental animal models exhibit high oxidative stress due to persistent and chronic hyperglycemia, thereby deplete the activity of the antioxidative defense system and thereby promote the generation of free radicals. The current study examined the effects of vitamin E on oxidative stress and membrane fluidity in the brain of diabetes-induced rats. METHODS: Sprague Dawley male rats were randomly assigned to normal and streptozotocin (STZ) induced diabetic groups. The diabetic groups were fed a vitamin E-free diet, 40 mg vitamin E/kg diet, or 400 mg vitamin E/kg diet. Diabetes was induced with STZ after 3 weeks of the experimental diet, then the rats were sacrificed 9 days later to determine the oxidative stress and cell membrane fluidity in the brain. RESULTS: Dietary vitamin E strengthened the antioxidative defense system with an increased activity of the antioxidant enzymes, such as superoxide dismutase (SOD) and glutathione peroxidase (GSH-px) and increased vitamin E content, in the brain of the diabetes-induced experimental rats. Accordingly, vitamin E was found to reduce the accumulation of reactive oxygen species (ROS), such as superoxide radical decrease the generation of oxidative damage substances, such as the carbonyl value, increase the membrane fluidity lowered by oxidative damage, and significantly improve the lipid composition. CONCLUSIONS: Vitamin E was found to be excellent for strengthening the antioxidative defense system, reducing the generation of ROS and damaging oxidative substances, and maintaining membrane fluidity in the brain of diabetes-induced rats. PMID- 14734203 TI - Practical measurement of total and bioavailable estradiol in female macaques. AB - BACKGROUND: Commercially available, direct assay kits provide a simple and rapid means for measuring estradiol. Similarly, ammonium sulfate precipitation of SHBG bound steroids is a reliable alternative to equilibrium dialysis or ultrafiltration for assessing bioavailability. However, while these techniques are useful for humans, they have yet to be evaluated systematically for macaques species which are frequently used to model estrogen effects on women's health. METHODS: A reference assay (which included chromatography) and two modified versions of a human kit (one in which monkey serum was assayed directly, and another in which an extraction step was added) were used to measure estradiol in matching samples. Ammonium sulfate precipitation and an established ultrafiltration technique were used to assess bioavailability. RESULTS: Values from both kit modifications correlated significantly with those from the reference assay. Although both modifications underestimated values, the addition of the extraction step resulted in far more useful estimates due to the consistency of underestimation across the standard curve. Measures of bioavailability from ammonium sulfate precipitation and ultrafiltration were strongly correlated and consistent across all concentrations. CONCLUSIONS: Simplified techniques developed for humans can be used in macaques, although the addition of an extraction step markedly improves the performance of direct assay kits. PMID- 14734204 TI - Clinical and molecular aspects of haemoglobinopathies in Tunisia. AB - BACKGROUND: For the last two decades, studies on the population genetics of Tunisians have focused on variations of protein and genetic markers. Results confirmed the genetic heterogeneity of Tunisians caused by the admixtures with migratory human groups arriving mainly from Africa, Europe, and Asia. These studies also allowed the screening of rare mutants and many haemoglobin variants. METHODS: The present study delineates the incidence of the different haemoglobinopathies in Tunisia. Previously collected data and results obtained from epidemiological and clinical studies of 1238 blood donors and 276 patients were compared. The chromosomal backgrounds of different haemoglobinopathies were explored by molecular techniques (denaturing gradient gel electrophoresis (DGGE), amplification refractory mutation system (ARMS) polymerase chain reaction (PCR), and sequencing). RESULTS: This study indicates that appropriate DNA methodologies required for a nationwide preventive program in Tunisia are available and that prenatal diagnosis is feasible. Additionally, analysis of sequence polymorphisms allowed a better understanding of the gene recombination events and their application for tracing back the origin and the diffusion of the mutations. CONCLUSIONS: Molecular analysis techniques such as DGGE and ARMS PCR are socially and economically the most suitable techniques to be used in Tunisia for the detection and the identification of haemoglobin abnormalities. At present, their use is essential to conduct a clear and efficient screening program. PMID- 14734205 TI - Fructosamine, glycated hemoglobin, and dietary carbohydrates. AB - BACKGROUND: Glycated hemoglobin (HbA(1c)), a marker of glycemia in the previous 3 months, was found to be associated with dietary saturated, fat but not with carbohydrates, in recent population surveys. Another nonenzymatically glycated substance in the blood, fructosamine, a marker of glycemia in the previous 3 weeks, is poorly correlated with HbA(1c) in nondiabetic subjects. The aim of this study is to compare the correlation of glycated hemoglobin and fructosamine with dietary carbohydrate intake in the same subjects. SUBJECTS AND METHODS: Seventy one individuals from a cohort study on diet and cancer entered this study. Serum fructosamine was measured by a standard colorimetric method, and glycated hemoglobin by high-performance liquid chromatography (HPLC). Diet was measured by a validated semiquantitative food frequency questionnaire. The correlation of fructosamine and glycated hemoglobin with dietary variables, corrected for calories, was evaluated by multiple correlation. RESULTS: Fructosamine was more strongly correlated with dietary sugar (r=0.26, p=0.05) than HbA(1c) was (r=0.001, p=0.99). Fructosamine was also inversely correlated with energy, and glycated hemoglobin with vitamin C. CONCLUSIONS: Fructosamine appears to be more related to dietary sugar intake than glycated hemoglobin and may be a marker of exposure to dietary carbohydrates, particularly simple sugars, in epidemiological studies. PMID- 14734206 TI - Procalcitonin levels in plasma in oncohaematologic patients with and without bacterial infections. AB - BACKGROUND: The flogosis markers currently in use show both low sensitivity and specificity, particularly in neoplastic and degenerative diseases. Procalcitonin (PCT) is a pro-peptide of calcitonin produced mainly but not only in the C-cells of the thyroid glands and, as several studies show, PCT levels in plasma increase during infections. Bacterial infections are also the main cause of death in oncological patients. Furthermore, in patients with leukaemia in chemotherapy recovery, infections often induce relapses. The aim of the present study is to detect PCT levels in plasma in oncohaematologic patients with and without infections. METHODS: The study was carried out on 54 patients by a quantitative automated immunoassay. RESULTS: PCT plasma levels > or =0.5 ng were detected in 27 out of 30 patients (90,0%) with bacterial infections; 8 out of 9 patients (88,9%) with viral infections and in 12 out of 15 patients in the control group without statistically significant differences. CONCLUSIONS: The results, which differ from those in the literature, are discussed. PMID- 14734207 TI - Ginkgo biloba prevents mobile phone-induced oxidative stress in rat brain. AB - BACKGROUND: The widespread use of mobile phones (MP) in recent years has raised the research activities in many countries to determine the consequences of exposure to the low-intensity electromagnetic radiation (EMR) of mobile phones. Since several experimental studies suggest a role of reactive oxygen species (ROS) in EMR-induced oxidative damage in tissues, in this study, we investigated the effect of Ginkgo biloba (Gb) on MP-induced oxidative damage in brain tissue of rats. METHODS: Rats (EMR+) were exposed to 900 MHz EMR from MP for 7 days (1 h/day). In the EMR+Gb groups, rats were exposed to EMR and pretreated with Gb. Control and Gb-administrated groups were produced by turning off the mobile phone while the animals were in the same exposure conditions. Subsequently, oxidative stress markers and pathological changes in brain tissue were examined for each groups. RESULTS: Oxidative damage was evident by the: (i) increase in malondialdehyde (MDA) and nitric oxide (NO) levels in brain tissue, (ii) decrease in brain superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities and (iii) increase in brain xanthine oxidase (XO) and adenosine deaminase (ADA) activities. These alterations were prevented by Gb treatment. Furthermore, Gb prevented the MP-induced cellular injury in brain tissue histopathologically. CONCLUSION: Reactive oxygen species may play a role in the mechanism that has been proposed to explain the biological side effects of MP, and Gb prevents the MP-induced oxidative stress to preserve antioxidant enzymes activity in brain tissue. PMID- 14734208 TI - The effect of class-specific protease inhibitors on the stabilization of B-type natriuretic peptide in human plasma. AB - BACKGROUND: B-type natriuretic peptide (BNP) is a cardiac hormone that regulates hemodynamic equilibrium. In the circulation, its activity is controlled by proteolytic factors. Accurate measurement of BNP in a patient's plasma may be affected by degradation due to proteolysis. OBJECTIVE: We report on the identification and performance of classes of protease inhibitors that stabilize BNP in plasma. DESIGN AND METHODS: Using the Bayer ADVIA Centaur BNP assay, we measured the effect of arginine, serine and/or specific kallikrein protease inhibitors (PIs) on exogenous spiked or endogenous BNP in patient plasma. RESULTS: Compared to controls without inhibitor, all PIs were capable, to varying degrees, of retarding the rate of proteolytic degradation. The kallikrein specific inhibitor, D-Phe-Phe-Arg-chloromethylketone (PPACK II) was most effective as a single constituent and was able to eliminate BNP degradation in patient samples for up to 6-10 days when stored at 2-8 degrees C. CONCLUSIONS: The stability of BNP was markedly increased in the presence of kallikrein specific PPACK II and a broad spectrum of serine PIs. Use of these compounds offers a simple method of extending sample handling and storage of plasma samples containing BNP. PMID- 14734209 TI - Polymorphism of interleukin-1 receptor antagonist gene and obesity. AB - BACKGROUND: Cytokines appear to be major regulators of adipose tissue metabolism. Interleukin-1 receptor antagonist (IL-1ra) serum concentrations are increased in human obesity, and are under strong genetic control. We tested the hypothesis that the IL-1ra gene might be a candidate for obesity. METHODS: We investigated the frequency of a penta-allelic 86-bp tandem repeat (VNTR) in the intron 2 of IL 1ra gene in 52 lean (body mass index (BMI) <25 kg/m(2)), 133 overweight (BMI 25 29.9 kg/m(2)) and 76 obese (BMI > or =30.0 kg/m(2)) otherwise healthy Korean women. Total fat mass and percent body fat were determined by dual-energy X-ray absorptiometry. Genomic DNA was extracted and used for polymerase chain reaction based genotyping of IL-1ra. RESULTS: Carriers of the allele 2 did not show a significant difference in physical and clinical characteristics. The genotypic, or allelic distribution did not differ markedly between the three groups. The relative risk of being obese in comparison with lean group tended to be higher in allele 2 carriers, but not significantly. CONCLUSIONS: We found no relationship between the IL-1ra polymorphism and BMI in Korean women. PMID- 14734210 TI - Poncirus trifoliata fruit induces apoptosis in human promyelocytic leukemia cells. AB - BACKGROUND: Substances inducing apoptosis have shown efficacy in the treatment of cancers. Poncirus trifoliata (L.) Raf. (Rutaceae) fruits (PTF) has been used for the treatment of various cancers among Korean Oriental Medical doctors. METHODS: PTF-induced cytotoxicity of human leukemia HL-60 cells was monitored by the MTT assay. The apoptosis was determined by (a) apoptotic morphology in microscopy; (b) DNA fragmentation in electrophoresis and FACS analysis; and (c) activation of caspase-3 and poly-ADP-ribose polymerase (PARP) cleavage assay. RESULTS: The cytotoxic activity of PTF in HL-60 cells was increased in a concentration- and time-dependent manner. PTF caused the cell shrinkage, cell membrane blebbing, apoptotic body and DNA fragmentation. PTF-induced apoptosis is accompanied by the activation of caspase-3 and the specific proteolytic cleavage of PARP. However, PTF did not show cytotoxicity in normal peripheral blood mononuclear cells. CONCLUSIONS: Our novel finding provides evidence that PTF could be a candidate as an anti-leukemic agent through apoptosis of cancer cells. PMID- 14734211 TI - Serum concentrations of dehydroepiandrosterone and dehydroepiandrosterone sulfate and their relation to cytokine production during and after normal pregnancy. AB - BACKGROUND: Since dehydroepiandrosterone (DHEA) and dehydroepiandrosterone sulfate (DHEAS) have been suggested to have immunoregulatory effects, changes in the levels of these substances during and after pregnancy might affect the maternal immune system. We examined serum concentrations of DHEA and DHEAS, and cytokine production during pregnancy and after delivery. METHODS: The subjects were 73 normal pregnant, 76 normal postpartum and 30 normal non-pregnant women. Whole-blood was stimulated with phorbol 12-myristate 13-acetate (PMA) and ionomycin and the levels of cytokines in the supernatant were measured using enzyme-linked immunosorbent assay (ELISA). DHEA and DHEAS were measured using ELISA and gas chromatography-mass spectrometry (GC-MS), respectively. RESULTS: The serum DHEA levels increased in the first and in the second trimesters and decreased after delivery until 11 months postpartum. DHEAS levels were decreased in the second and in the third trimesters and returned to non-pregnant levels after pregnancy. All measured cytokines (IFN-gamma, IL-2, IL-4 and IL-10) were decreased during pregnancy and subsequently increased postpartum. We found significant negative correlations between DHEA and cytokine levels. CONCLUSIONS: Increase of serum DHEA in the first and the second trimesters may suppress immune reaction during pregnancy, while a decrease of DHEA after delivery may induce postpartum enhancement of the maternal immune system. DHEA may be involved in modifying the maternal immune responses during and after pregnancy. PMID- 14734212 TI - Comparing the ESA HPLC total homocysteine assay with electrochemical detection to the CDC in-house HPLC assay with fluorescence detection. AB - BACKGROUND: Environmental science has developed a simple high performance liquid chromatography (HPLC) assay with electrochemical detection for total homocysteine (tHcy) measurement that does not require derivatization of free thiols. We evaluated this method and compared it with the CDC HPLC assay with fluorescence detection (FD). METHODS: tHcy is measured after reduction of disulfides/protein bound thiols and protein precipitation using four channels of an ESA CoulArray detector. L-homocystine is used as calibrator, penicillamine as internal standard. RESULTS: Aqueous calibration of the ESA assay resulted in overestimation of tHcy by approximately 30% compared to the HPLC-FD method. Calibration in plasma alleviated the matrix effect. The within- (n=3) and between run (n=20) imprecision was <6%, the linearity up to 100 micromol/l was excellent, and the recovery of tHcy added to plasma was nearly complete (98.7%+/-2.3%). Good correlation was observed between both methods for 266 plasma samples. The ESA assay showed a minimal negative bias of 0.28 micromol/l (3.3%). CONCLUSION: The ESA tHcy assay performed well in terms of accuracy and precision, and showed good agreement with the CDC HPLC-FD assay when calibrated in plasma. The major advantage of this assay is that it does not require sample derivatization. Disadvantages include instability of the prepared samples for prolonged storage and matrix effects. PMID- 14734213 TI - Apolipoprotein E genotyping in the Malay, Chinese and Indian ethnic groups in Malaysia-a study on the distribution of the different apoE alleles and genotypes. AB - BACKGROUND: Apolipoprotein E (apoE) is encoded by a polymorphic gene located on chromosome 19. The three common apoE alleles are epsilon2, epsilon3 and epsilon4. We studied the frequencies of the apoE alleles and genotypes in the three ethnic groups-Malay, Chinese and Indian-in Malaysia using DNA amplification followed by agarose gel electrophoresis. METHODS: EDTA blood was collected and DNA was extracted using proteinase K-SDS digestion and purified by phenol-chloroform extraction. The apoE gene sequence was amplified using the PCR and apoE genotyping was performed by restriction enzyme digestion with HhaI. RESULTS: Genotyping of the apoE gene produces six genotypes-E2/E2, E2/E3, E3/E3, E2/E4, E3/E4 and E4/E4. The most common apoE genotype in the Malays, Chinese and Indians studied was E3/E3, thus the most common apoE allele was epsilon3. The three common apoE genotypes were E3/E3 followed by E3/E4 and E2/E3, except in the Indians where E2/E3 was not detected. The three apoE alleles were confirmed in the Malays, Chinese and Indians except for the epsilon2 allele which was absent in the Indians. CONCLUSION: The combined frequency of the apoE alleles in the Malays, Chinese and Indians was 0.058, 0.829 and 0.114 for epsilon2, epsilon3 and epsilon4, respectively. PMID- 14734214 TI - Redox status and lipid peroxidation in alcoholic hypertensive patients and alcoholic hypertensive patients with diabetes. AB - BACKGROUND: Free radical-mediated oxidative stress has been implicated in the progression of alcoholic hypertension and diabetic hypertension. METHODS: The lipid peroxides and antioxidant status of plasma and erythrocytes were investigated in alcoholic hypertensive patients and alcoholic hypertensive patients with diabetes and compared with normal subjects. RESULTS: A significant increase is observed in the levels of glucose, lipid peroxidation (P<0.05) in the alcoholic hypertensive patients with/without diabetes and the increase was significantly higher in alcoholic hypertensive patients with diabetes. The activities of erythrocyte antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and reduced glutathione (GSH) and plasma concentrations of GSH, vitamin C, vitamin E and beta-carotene decreased significantly and the level of ceruloplasmin increased in alcoholic hypertensive patients with/without diabetes when compared to normal subjects. Plasma GSH and vitamin E levels exhibited a further decrease in alcoholic hypertensive patients with diabetes. CONCLUSIONS: An enhanced lipid peroxidation is observed in alcoholic hypertensive patients with diabetes and a more pronounced decrease in the levels of plasma GSH and vitamin E among antioxidants. PMID- 14734215 TI - Performance characteristics of the IMMUNLITE 2000 erythropoietin assay. AB - BACKGROUND: Erythropoietin (EPO) is a glycoprotein hormone produced by specialized cells in the kidney that acts as the primary regulator of erythropoiesis. Serum EPO measurements are useful for diagnostic evaluations of anemia and polycythemia. METHODS: The IMMULITE 2000 is an automated random access immunoassay analyzer for the central laboratory. The limit of detection (LOD), linearity, imprecision, comparison to another commercially available assay, and reference interval of a new EPO assay for this analyzer were assessed. RESULTS: The LOD was 0.2 U/l. The assay was linear within an allowable systematic error of 10% over the range tested (2-178 U/l). The total imprecision of the new assay was <7% for concentrations from 15.8 to 68.4 U/l. Comparison with the Advantage EPO assay method showed comparable results. Deming regression analysis gave a slope of 1.13+/-0.02, an intercept of -1.09+/-0.97, and a S(y/x) of 7.1 (r=0.99) over the range tested (2-200 U/l). CONCLUSIONS: The IMMULITE 2000 EPO assay shows acceptable performance and is suitable for routine clinical use. PMID- 14734216 TI - A sensitive colorimetric assay for polyamines in erythrocytes using oat seedling polyamine oxidase. AB - BACKGROUND: Most of the polyamines circulating in blood are spermidine (Spd) and spermine (Spm) with only trace amounts of putrescine (Put), and they are mainly localized in erythrocytes. We developed a simple and sensitive colorimetric assay for polyamines in erythrocytes using oat seedling polyamine oxidase (OSPO). The method is based on the unique substrate specificity of OSPO, which is active toward Spd and Spm, but not toward diamines such as Put and cadaverine and monoamines such as histamine. METHODS: The polyamines, which were purified from packed erythrocytes by weak cation-exchange chromatography, were incubated with OSPO at 37 degrees C for 15 min. In the presence of the H(2)O(2) produced by this polyamine oxidase reaction and a new chromogen, N-(carboxymethylaminocarbonyl) 4,4'-bis(dimethylamino)-diphenylamine sodium salt (DA-64), peroxidase (POD) catalyzes the formation of N-[4-[[4-(dimethylamino)phenyl]imino]-2,5 cyclohexadien-1-ylidene]-N-methylmethanaminium chloride (Bindschedler's Green) having an absorption maximum at 727 nm. RESULTS: The detection limit was 0.2 microM/l for packed erythrocytes. The within-run and between-run precisions (coefficient of variation, CVs) were 5.6-15.2% and 6.5-16.4%, respectively. Analytical recoveries were 93.3-97.4%. Polyamine values obtained by this assay correlated well with those obtained by an HPLC (y=0.948x + 1.912; r=0.944; n=46). CONCLUSIONS: This colorimetric assay is simple and highly sensitive and practical for clinical use. PMID- 14734217 TI - Antioxidant potential of vitamins A, E and C in modulating oxidative stress in rat brain. AB - BACKGROUND: Stress is known to affect synaptic plasticity, dendritic morphology and induces neurotoxic damage in humans, probably through generation of free radicals. Both ex vivo antioxidant vitamins and in vivo free radical scavenging enzymes exist. In the present study, restraint stress induced pro-oxidant status of rat brain was evaluated in terms of measurement of glutathione (GSH), lipid peroxidation (thiobarbituric acid reactive substances, TBARS) and free radical scavenging enzymes activities. The efficacy of antioxidant vitamins A, E and C alone and in combination was also evaluated in modulating inherent antioxidant system in stressed rats. METHODS: Rats were treated with vit A, E and C alone (15 mg/kg of body weight) and in combination vitamins (E and C) prior to and after 6 h of restraint stress exposure. Both nonstressed and stressed rats were handled simultaneously. Pro-oxidant status of brain tissue was evaluated by determining the levels of GSH, TBARS and activities of superoxide dismutase (SOD), glutathione-S-transferase (GST) and catalase (CAT). RESULTS: Restraint stress induced a decrease in the level of GSH and the activities of SOD, GST and catalase, while the levels of TBARS were found elevated. Both pre-stress and post stress vitamin treatments (either alone or combined) resulted in alteration of these parameters towards their controls values with a relative dominance by latter. Vitamin E was found most effective in restoring inherent antioxidant system, no additive effect was observed in combined vitamin treatment as expected. CONCLUSION: Immobilization of rats generated oxidative stress in rat brain, by decreasing the activities of SOD, GST, catalase and glutathione levels, while increasing the lipid peroxidation. Post stress vitamin E treatment was found most effective than vitamins A and C in enhancing the levels of glutathione and activities of SOD, GST and catalase and decreasing lipid peroxidation. Thus vitamin E can be given as a nutritional supplement for scavenging free radical generated in the brain tissues in order to reduce oxidative stress. PMID- 14734218 TI - Plasma homocysteine reference values of adult Malaysians from three ethnic groups. PMID- 14734219 TI - Inversion of chromosome 12 and lineage promiscuity in hematologic malignancies. AB - Rearrangements of the short arm of chromosome 12 are among the most common aberrations found in hematologic malignancies, including myelodysplastic syndromes, acute myelocytic leukemias, acute lymphoblastic leukemias, and non Hodgkin lymphomas. We report on a group of 46 patients with a variety of myelocytic and lymphoid malignancies, all with an inversion of chromosome 12. Both pericentric and paracentric inversions occurred. The identified hotspots for breakage were p13 and q24. These correspond to gene-rich areas of known chromosome instability. The inv(12) is difficult to detect and may be misinterpreted as a partial deletion by routine cytogenetics. Fluorescence in situ hybridization studies revised the G-banding interpretations of a deleted 12p in some cases to an inversion. The inv(12) may occur as the sole abnormality in both myelocytic and lymphoid malignancies, suggesting lineage promiscuity as seen with MLL and ETV6 gene disruptions. The majority of patients with the inv(12) had complex karyotypic changes that predicted a poor prognosis. Of the 24 patients with known clinical follow-up, many were refractory to chemotherapy and overall survival was short. PMID- 14734220 TI - Challenging colonic polyposis pedigrees: differential diagnosis, surveillance, and management concerns. AB - Hereditary polyposis syndromes show extensive phenotypic and genotypic heterogeneity within and among families, a situation that may hinder diagnosis. In these settings, germline mutation testing may be the sine qua non for diagnosis if such a mutation is identified in a patient or family. We provide examples of phenotypically differing polyposis pedigrees depicting various challenges in hereditary polyposis syndrome diagnosis. Our purpose is to augment physician understanding of phenotypic variation and thus help identify high-risk presymptomatic family members who could benefit from highly targeted surveillance and management strategies. We describe nine familial polyposis pedigrees displaying anecdotal clinical problems that can confound the differential diagnosis. Emphasis was given to a multidisciplinary approach focusing on pathological confirmation with respect to number, histology, and location of polyps in the gastrointestinal tract; a detailed family history of cancer at all anatomic sites; noncancer phenotypic features of hereditary polyposis syndromes; and appropriate molecular genetic testing in concert with genetic counseling. Improved physician understanding of the clinical natural history features, genetic transmission patterns, and appropriate gene testing will help in diagnosis and, ultimately, surveillance and management for the various hereditary polyposis syndromes. PMID- 14734221 TI - Overexpression of the BCL2 gene in a Sertoli-Leydig cell tumor of the ovary: a pathologic and cytogenetic study. AB - A case of virilizing ovarian Sertoli-Leydig cell tumor overexpressing the BCL2 gene and including a novel clonal chromosomal rearrangement of chromosome 18, der(5)t(5;18)(p13;q12),+6,+12, der(18)r(5;18)(p15.3p13;p11.3q12) is described. Further studies of these rare tumors are necessary to ascertain the significance of the findings. PMID- 14734222 TI - Association of loss of 1p and alterations of chromosome 14 in meningioma progression. AB - Meningiomas are usually benign tumors; however, they can recur after surgical resection and occasionally show histologic progression to a higher grade II and III malignancy. The second most frequently reported genetic abnormality after 22q loss is deletion of 1p, although alterations in 9q, 10q, and 14q are also implicated in meningioma progression. Fourteen tumors comprising six benign, four atypical, and four malignant meningiomas were examined by means of cytogenetic and fluorescence in situ hybridization analysis. All tumors showed losses in different regions of 1p, with 1p11, 1p13, 1p21, 1p22, 1p32, and 1q21 breakpoints; eight tumors also presented alterations of chromosome 14. Five of the six cases with deletions on 1p and normal chromosome 14 were grade I, and two were recurrent. All but one of the eight cases with simultaneous 1p deletion and alterations of chromosome 14 were grade II (3 cases) and grade III (4 cases); all the grade III cases were recurrent. These results support the possible association between changes in 1p and chromosome 14 with the evolution of aggressive meningiomas through tumor progression. PMID- 14734223 TI - Molecular cytogenetic characterization of a case of Mullerian adenosarcoma. AB - Mullerian adenosarcoma is a distinctive type of mixed Mullerian tumor of the female genital tract. To our knowledge, no cytogenetic data have been documented on Mullerian adenosarcoma in the literature so far. We report here the chromosomal findings of a Mullerian adenosarcoma in a 15-year-old female. Cytogenetic and molecular cytogenetic analysis revealed a complex karyotype involving chromosomes 2, 8, 10, 13, 19, and 21. These numerical and structural abnormalities may be of etiologic significance. This report may highlight the potential value of molecular cytogenetic analysis in differential diagnosis of Mullerian tumors. More cases are warranted to further genetically characterize this type of neoplasm. PMID- 14734224 TI - Uncommon cytogenetic findings in a case of splenic marginal zone lymphoma with aggressive clinical course. AB - The majority of splenic marginal zone lymphoma (SMZL) patients experience an indolent clinical course; however, some cases transform to a high-grade lymphoma. Cytogenetic analyses have shown that chromosome 7 is the most frequently altered chromosome and, in some cases, 7q deletion has been found as a single aberration, suggesting its association with the development of SMZL. We studied one patient showing clinical features of SMZL with an aggressive course. Immunophenotypic, conventional and molecular cytogenetic techniques were applied to support the diagnosis. The immunophenotype of peripheral blood mononuclear cells showed the presence of 90% B-lymphocytes. Cytogenetic analysis indicated the presence of a stem-line lacking normal chromosomes 7, but showing a der(7) and a ring, and a side-line with additional aberrations: t(2;22), add(8). Fluorescence in situ hybridization analysis revealed a loss of the 7q32 region. Nonclonal rearrangements involving chromosome 7 were also detected. Chromosome 7 rearrangements were studied to investigate their evolution during the development of the pathology. We have shown that in this patient both chromosomes 7 underwent different rearrangements leading to a loss of the 7q32 region and that the ring chromosome originated from chromosome 7 and was associated with a t(7;7) (p22;q31). We conclude that not only the 7q deletion but also the proneness of chromosome 7 to rearrange might have played a role in the progression of this SMZL. PMID- 14734225 TI - A chronic myelocytic leukemia case bearing deletions on the three chromosomes involved in a variant t(9;22;11). AB - Genomic deletions on the derivative chromosomes bearing the reciprocal fusion gene have recently been reported in chronic myelocytic leukemia (CML). We here describe a CML case with a variant rearrangement t(9;22;11)(q34;q11;q13) showing the loss of chromosome 11 sequences in addition to der(9) deletions. Known tumor suppressor genes involved in apoptosis and in the control of cell proliferation were found to be mapped to the lost sequences. Our findings indicate that genomic deletions may occur also on the third derivative chromosome in variant t(9;22). PMID- 14734226 TI - Interphase cytogenetic analysis of clonality in peripheral blood cells from a patient with Down syndrome and acute megakaryoblastic leukemia. AB - A combination of fluorescence-activated cell sorting and interphase fluorescence in situ hybridization (FISH) techniques was used to detect a clonal chromosomal marker in blasts, granulocytes, and T and B lymphocytes of the peripheral blood from a patient with Down syndrome and acute megakaryoblastic leukemia (AMKL) associated with trisomy 8 as a karyotypic abnormality. Immunophenotypic studies with flow cytometry showed two populations of leukemic blasts distinguished by their expression of the CD34 antigen. Interphase FISH studies revealed clonal trisomy 8 FISH signals in almost all blast cells, regardless of CD34 expression, as well as in a small subpopulation of granulocytes. Normal chromosome 8 signal patterns were detected in T and B cells and in a great majority of granulocytes. The present study provides evidence for the clonal involvement of leukemic blasts in AMKL of Down syndrome, indicating that a trisomy 8 abnormality may be a primary event in leukemogenesis. The transformation occurs in progenitor cells with limited myeloid differentiation and without involvement of lymphoid lineage cells. PMID- 14734227 TI - Cardiac myxoma: a cytogenetic study of two cases. AB - Two cases of cardiac myxoma, each arising in the left atrium, are presented. One tumor contained the clonal abnormality i(17)(q10),der(20)t(1;20)(q21;q11.2) and the second tumor contained add (9)(p22),+12. Such rearrangements have not been previously reported in these tumors. PMID- 14734228 TI - Methylation status of TP73 in meningiomas. AB - Deletions at 1p are frequent in meningioma and represent a genetic marker associated with the genesis of atypical WHO grade II forms. Previous mutational analysis of TP73, a structurally and functionally TP53 homologous gene located at 1p36.33, failed to demonstrate a significant rate of sequence variations linked to gene inactivation in meningiomas with 1p loss. As an alternative, TP73 may be inactivated through aberrant 5' CpG island methylation, a primary mechanism participating in the inactivation of tumor suppressor genes during tumorigenesis. We determined the methylation status of the TP73 gene in a series of 60 meningiomas (33 grade I, 24 grade II, and 3 grade III samples), including tumors with deletion at 1p (n=30) and with intact 1p (n=30). Aberrant methylation was detected in 10 cases (33%) with 1p deletion and in 3 tumors (10%) with retention of alleles at this chromosome arm. The distribution of the 13 cases of methylation according to malignancy grade was 7 grade I, 5 grade II, and 1 grade III tumor. Accordingly, although TP73 aberrant methylation was more frequent in meningiomas with 1p deletion (P<0.05), no association with the grade of malignancy could be established. These findings, together with the previously reported increased TP73 expression in malignant meningiomas suggest that opposing functions of this gene may characterize distinct subsets of tumors: suppressed or reduced expression as a result of CpG methylation in some grade I-grade II tumors, and enhanced expression in some more malignant forms. PMID- 14734229 TI - A novel t(6;7)(p24;q21) in a chronic myelocytic leukemia in complete cytogenetic remission after therapy with imatinib mesylate. AB - Emergence of additional cytogenetic clones in chronic myelocytic leukemia (CML) patients who become Philadelphia chromosome-negative (Ph-) after alpha-interferon therapy (or more recently with imatinib mesylate) have been described. We report here a case of a novel t(6;7)(p21;q23) that developed in a CML patient in complete cytogenetic remission during imatinib therapy. In this case, fluorescence in situ hybridization and reverse transcriptase polymerase chain reaction showed a normal pattern for BCR and ABL genes, suggesting that a different and unrelated clone developed after the disappearance of the Ph chromosome. PMID- 14734230 TI - Familial myelodysplastic syndromes, monosomy 7/trisomy 8, and mutator effects. AB - A family is reported, in which two sisters presented with myelodysplastic syndrome (MDS), namely refractory anemia with excess of blasts in transformation (RAEB-t), and refractory anemia (RA). Bone marrow chromosome changes were present in both: trisomy and tetrasomy 8 (with a pericentric inversion of one chromosome 8) in the older sister, and monosomy 7 (with clones with additional trisomies 19 and 21) in the younger one. Molecular data were obtained on the parental chromosome involved in these numerical anomalies, which proved to be of paternal origin in these cases. The observations of this family, and a review of familial cases of MDS/acute myeloid leukemia (AML), led us to consider that they may be divided into two groups: those which arise on the basis of a Mendelian predisposing disorder exerting a mutator effect, often with the acquisition of monosomy 7, and those in which no specific Mendelian predisposing disease is recognized, as the familial monosomy 7 cases and the one reported here. We postulate that in these families an inherited mutator effect is present and that it causes a karyotype instability, which leads to MDS/AML, often through the acquisition of monosomy 7 and trisomy 8. PMID- 14734231 TI - Is trisomy 5 a distinct cytogenetic subgroup in acute lymphoblastic leukemia? AB - Acute lymphoblastic leukemia (ALL) is characterized by recurrent clonal chromosomal abnormalities, with numerical abnormalities being a common feature especially among children. Case reports in the literature suggest that one such recurrent numerical abnormality is the gain of chromosome 5 (trisomy 5) as the sole abnormality; due to the rarity of these cases, however, little is known about their incidence, clinical features, and prognosis. We have identified seven cases with trisomy 5 as the sole or primary chromosomal abnormality from a total of 3,400 karyotypes collected in the Leukaemia Research Fund UK Cancer Cytogenetics Group Karyotype Database. All cases had a precursor B-cell immunophenotype and there was a male predominance. Five patients were children aged between 7 and 14 years old. Four of the six patients with a reasonable follow-up period had relapsed, indicating a poor prognosis. We conclude that trisomy 5 as the sole numerical abnormality occurs predominantly in older children, may be associated with a poor outcome, and may represent a distinct, albeit rare, cytogenetic subgroup in ALL. PMID- 14734232 TI - Microsatellite instability is a rare finding in tumors of patients with both primary renal and rectal neoplasms. AB - Multiple primary malignancies in an individual patient are thought to be a common feature of hereditary disease. Asymptomatic renal neoplasms have been described in up to 4% of rectal cancer patients. We have assessed whether microsatellite instability is present in the rectal and renal tumors of patients presenting at our institution with primary renal and rectal cancers. Of the seven patients presenting with both diagnoses, paraffin blocks were available from five cases of colorectal cancer and six renal neoplasms. Five of six cases of renal tumors were informative and all were microsatellite instability (MSI) stable. One renal tumor was deemed inconclusive because of inadequate amplification. Four of five cases of rectal cancer were MSI stable. These data suggest that these renal and rectal tumors developed independently of a mismatch repair defect and that, unlike epithelial tumors of the renal collecting system, renal parenchymal lesions are unlikely to be part of the spectrum of hereditary nonpolyposis colorectal cancer. PMID- 14734233 TI - Secondary chromosomal changes in 34 Philadelphia-chromosome-positive chronic myelocytic leukemia patients from the Mexican West. AB - The clonal evolution in t(9;22)-positive chronic myelocytic leukemia (CML) is well established. Four major changes occur in more than 70% of patients: +8, i(17q), +19, and an extra Philadelphia chromosome. The frequencies of secondary chromosomal changes in 34 patients from the states of Jalisco, Nayarit, Michoacan, and Colima (the Mexican West) with Philadelphia-chromosome-positive CML were assessed. The most frequent abnormalities were tetraploidy (12 cases); +8, inv(3)(q21q26), and octoploidy (3 cases each); and +der(22)(2 cases). Some translocations not previously associated with CML were observed, such as t(2;7)(p12;q36), t(3;6)(q26;p25), t(3;17)(q26;p13), and t(6;17)(q21;q23 approximately q25). Significant differences were found for +8 with respect to population results from Japan and from southern, eastern, and western Europe; for i(17)(q10) from eastern Europe; for +19 from Japan and western Europe; and for +der(22) from Japan, southern Europe, and western Europe. Although polyploidy could result from endomitosis, there is no direct evidence that the BCR/ABL protein influences such a process; however, protein kinases such as MAPK, which are involved in endomitosis, are activated by the BCR/ABL protein, and so the BCR/ABL protein could promote endomitosis through the MAPK pathway. PMID- 14734234 TI - Development and progression of a Philadelphia-chromosome-negative acute myelocytic leukemia clone in a patient with Philadelphia-chromosome-positive chronic myelocytic leukemia. AB - We report a patient with typical Philadelphia-chromosome-positive chronic myelocytic leukemia who developed Philadelphia-chromosome-negative acute myelocytic leukemia following autologous stem cell transplantation. The implications of this observation for disease monitoring and treatment are discussed. PMID- 14734235 TI - Trisomies 14 and 22 in acute myelocytic leukemia with multilineage dysplasia. PMID- 14734236 TI - Rapid diagnosis of acute promyelocytic leukemia (PML): applicability of flow cytometry and PML protein immunofluorescence. PMID- 14734237 TI - A new case of t(11;17)(q23;q21) with MLL rearrangement. PMID- 14734238 TI - Attentional load associated with performing and stabilizing a between-persons coordination of rhythmic limb movements. AB - This study addressed the issue of intentional stabilization of between-persons coordination patterns (in-phase/isodirectional and anti-phase/non-isodirectional) and the attentional cost incurred by the nervous system in maintaining and further stabilizing these coordination patterns. Five pairs of participants performed in-phase and anti-phase interpersonal coordination patterns in dual task conditions (coordination+RT task). Results showed that: (1) isodirectional pattern (in-phase) was more stable than non-isodirectional pattern (anti-phase), (2) both iso- and non-isodirectional pattern were stabilized intentionally, (3) RT was lower for the isodirectional pattern (i.e., the most stable), and (4) attentional manipulation led to a trade-off between pattern stability and RT performance. These results suggest that performing between-persons coordination patterns incurs a central cost that depends on the coupling strength between the limbs. These findings are consistent with the previous studies in intrapersonal coordination. PMID- 14734239 TI - Patients suffering from nonspecific work-related upper extremity disorders exhibit insufficient movement strategies. AB - The purpose of this research was to investigate whether patients experiencing nonspecific complaints of the forearm caused by sustained use of the personal computer exhibit deviant movement strategies as compared to healthy participants. Patients (N=10) and controls (N=24) performed a graphical aiming task combined with an auditory memory task. Force production (pen pressure), kinematic- and performance variables were recorded. During a trial, the control group gradually increased pen pressure from the stationary phases to the dynamic phase. The patients increased their pen pressure much more abruptly and to such a degree that the final pressure during real-time movement far exceeded that of the controls. Memory load led to a greater increase of pen pressure from the stationary phase to the dynamic phase in the patient group. Patients further displayed longer reaction times. The results are discussed within the framework of our recent theory on the role of neuromotor noise in the regulation of task performance under conditions of stress. PMID- 14734240 TI - Mental effort causes vigilance decrease due to resource depletion. AB - The resource view on vigilance performance was tested. First, a low demanding task was compared with a similar low demanding task in which stimulus presentation was less monotonous due to added, irrelevant, stimuli. The resource view, maintaining that vigilance is lowered by hard mental work, predicts that addition of irrelevant stimuli will not affect performance. The classic arousal theory, however, states that arousal drops due to monotonous stimulus presentation and predicts that decreasing monotony will enhance performance. Results showed that performance was unaffected by added stimuli. Second, we tested whether a high-demanding task (with identical stimulus presentation as the low demanding task, but different instruction) would cause a greater decline in performance than the low demanding task. Indeed, in the high-demanding task performance was affected most. In sum, it appears that vigilance decreases due to hard mental work, which requires many resources. Both overall performance and decrement in performance can be explained in terms of resources, and this suggests that vigilance tasks should be resource-demanding tasks, which do not have to be of long duration. PMID- 14734241 TI - Age of acquisition and the cumulative-frequency hypothesis: a review of the literature and a new multi-task investigation. AB - Early-acquired words are processed faster than late-acquired words. This is a well-accepted effect within the word recognition literature. Different explanations have been proposed, either localizing the effect of age of acquisition (AoA) in a particular substage of word processing or seeing it as the result of the way in which information is stored and accessed in the brain in general. The cumulative-frequency hypothesis is an example of the latter type of explanation: it states that the total number of times a system has come across a particular stimulus will determine the speed with which the stimulus can be recognized. The present multi-task investigation provides a critical test of the different explanations. Results show that in a variety of word processing tasks the effects of frequency and AoA are highly correlated, and that the impact of AoA is consistently higher than would be expected on the basis of the cumulative frequency hypothesis. The findings are interpreted as evidence for recent demonstrations of a loss of plasticity in neural networks due to training and/or for mathematical models that describe the growth of the lexico-semantic network as the attachment of new nodes to existing nodes. PMID- 14734242 TI - Object concepts and action: extracting affordances from objects parts. AB - Two experiments with a part-generation task show that rated salience and production order of parts in artifacts are first predicted by their relevance for canonical actions, but also that they vary, depending on the current situation. In three further experiments participants read sentences describing actions (e.g., 'The woman shares the orange') followed by objects' parts from which it was easy or not to extract affordances (e.g., 'slice' vs. 'pulp'). They had to perform a part verification task or to evaluate whether or not the combination made sense. Parts from which it was easy to derive affordances were processed earlier and the combination was evaluated as the one which made more sense. Overall, results support the view that sensory-motor simulations underlie conceptualization and that concepts are action-based. PMID- 14734243 TI - Non-aqueous phase liquid spreading during soil vapor extraction. AB - Many non-aqueous phase liquids (NAPLs) are expected to spread at the air-water interface, particularly under non-equilibrium conditions. In the vadose zone, this spreading should increase the surface area for mass transfer and the efficiency of volatile NAPL recovery by soil vapor extraction (SVE). Observations of spreading on water wet surfaces led to a conceptual model of oil spreading vertically above a NAPL pool in the vadose zone. Analysis of this model predicts that spreading can enhance the SVE contaminant recovery compared to conditions where the liquid does not spread. Experiments were conducted with spreading volatile oils hexane and heptane in wet porous media and capillary tubes, where spreading was observed at the scale of centimeters. Within porous medium columns up to a meter in height containing stagnant gas, spreading was less than ten centimeters and did not contribute significantly to hexane volatilization. Water film thinning and oil film pinning may have prevented significant oil film spreading, and thus did not enhance SVE at the scale of a meter. The experiments performed indicate that volatile oil spreading at the field scale is unlikely to contribute significantly to the efficiency of SVE. PMID- 14734244 TI - Coupled modeling of cement/claystone interactions and radionuclide migration. AB - The interactions between cement and a clayey host-rock of an underground repository for intermediate-level radioactive waste are studied with the reactive transport code HYTEC for supporting performance assessment. Care is taken in using relevant time scales (100,000 years) and dimensions. Based on a literature review, three hypotheses are considered with respect to the mineralogical composition of the claystone and the neo-formed phases. In the long term, the pH is buffered for all hypotheses and important mineral transformations occur both in cement and the host-rock. The destruction of the primary minerals is localized close to the cement/claystone interface and is characterized by the precipitation of secondary phases with retention properties (illite, zeolite). However, beyond the zone of intense mineral transformations, the pore water chemistry is also disturbed over a dozen meters due to an attenuated but continuous flux of hydroxyl, potassium and calcium ions. Four interdependent mechanisms control the profile in the whole system: diffusion of the alkaline plume, mineralogical buffering, ion exchange and clogging of the pore space at the cement/claystone interface. The migration of a selected group of radionuclides (Cs, Ra, Tc and U) is explicitly integrated in the simulations of the strongly coupled system. Theoretical profiles of distribution coefficient (Kd) and solubility limit values are derived from the simulations, and their sensitivity with respect to the system evolution is estimated. PMID- 14734245 TI - In situ anion diffusion experiments using radiotracers. AB - Diffusion experiments in compacted bentonite have been carried out in situ using the borehole laboratory CHEMLAB. The "ordinary" anion iodide and the redox sensitive pertechnetate ion have been investigated. In spite of strongly reducing groundwater conditions, technetium was found to diffuse mostly unreduced as TcO4 , although in some spots in the compacted clay, the activity was significantly higher, which may be explained by reduction of some TcO4- by iron-containing minerals in the bentonite. The measured concentration profiles in the clay cannot be accommodated by assuming one single diffusion process. The experimental data are modeled assuming two diffusion paths, intralamellar diffusion and diffusion in external water. The apparent diffusivity for the intralamellar diffusion was found to be 8.6 x 10(-11) m2 s(-1) for iodide with a capacity factor of 0.1, while the apparent diffusivity for the diffusion in external water was found to be 5 x 10(-14) m2 s(-1) with alpha=2.26. The corresponding values for Tc were found to be Da= 6 x 10(-11) m2 s(-1), alpha=0.1 and Da= 1 x 10(-13) m2 s(-1), alpha=0.46, respectively. The diffusion constants and capacity factors obtained in this study are in accordance with data from laboratory experiments. PMID- 14734246 TI - Modeling of non-reactive solute transport in fractured clayey till during variable flow rate and time. AB - Fractures and biopores can act as preferential flow paths in clay aquitards and may rapidly transmit contaminants into underlying aquifers. Reliable numerical models for assessment of groundwater contamination from such aquitards are needed for planning, regulatory and remediation purposes. In this investigation, high resolution preferential water-saturated flow and bromide transport data were used to evaluate the suitability of equivalent porous medium (EPM), dual porosity (DP) and discrete fracture/matrix diffusion (DFMD) numerical modeling approaches for assessment of flow and non-reactive solute transport in clayey till. The experimental data were obtained from four large undisturbed soil columns (taken from 1.5 to 3.5 m depth) in which biopores and channels along fractures controlled 96-99% of water-saturated flow. Simulating the transport data with the EPM effective porosity model (FRACTRAN in EPM mode) was not successful because calibrated effective porosity for the same column had to be varied up to 1 order of magnitude in order to simulate solute breakthrough for the applied flow rates between 11 and 49 mm/day. Attempts to simulate the same data with the DP models CXTFIT and MODFLOW/MT3D were also unsuccessful because fitted values for dispersion, mobile zone porosity, and mass transfer coefficient between mobile and immobile zones varied several orders of magnitude for the different flow rates, and because dispersion values were furthermore not physically realistic. Only the DFMD modeling approach (FRACTRAN in DFMD mode) was capable to simulate the observed changes in solute transport behavior during alternating flow rate without changing values of calibrated fracture spacing and fracture aperture to represent the macropores. PMID- 14734247 TI - A cation exchange model to describe Cs+ sorption at high ionic strength in subsurface sediments at Hanford site, USA. AB - A theoretical and experimental study of cation exchange in high ionic strength electrolytes was performed using pristine subsurface sediments from the U.S. Department of Energy Hanford site. These sediments are representative of the site contaminated sediments impacted by release of high level waste (HLW) solutions containing 137Cs+ in NaNO3 brine. The binary exchange behavior of Cs+-Na+, Cs+ K+, and Na+-K+ was measured over a range in electrolyte concentration. Vanselow selectivity coefficients (Kv) that were calculated from the experimental data using Pitzer model ion activity corrections for aqueous species showed monotonic increases with increasing electrolyte concentrations. The influence of electrolyte concentration was greater on the exchange of Na+-Cs+ than K+-Cs+, an observation consistent with the differences in ion hydration energy of the exchanging cations. A previously developed two-site ion exchange model [Geochimica et Cosmochimica Acta 66 (2002) 193] was modified to include solvent (water) activity changes in the exchanger phase through application of the Gibbs Duhem equation. This water activity-corrected model well described the ionic strength effect on binary Cs+ exchange, and was extended to the ternary exchange system of Cs+-Na+-K+ on the pristine sediment. The model was also used to predict 137Cs+ distribution between sediment and aqueous phase (Kd) beneath a leaked HLW tank in Hanfordd's S-SX tank using the analytical aqueous data from the field and the binary ion exchange coefficients for the pristine sediment. The Kd predictions closely followed the trend in the field data and were improved by consideration of water activity effects that were considerable in certain regions of the vadose zone plume. PMID- 14734248 TI - Operator-splitting procedures for reactive transport and comparison of mass balance errors. AB - Operator-splitting (OS) techniques are very attractive for numerical modelling of reactive transport, but they induce some errors. Considering reactive mass transport with reversible and irreversible reactions governed by a first-order rate law, we develop analytical solutions of the mass balance for the following operator-splitting schemes: standard sequential non-iterative (SNI), Strang splitting SNI, standard sequential iterative (SI), extrapolating SI, and symmetric SI approaches. From these analytical solutions, the operator-splitting methods are compared with respect to mass balance errors and convergence rates independently of the techniques used for solving each operator. Dimensionless times, NOS, are defined. They control mass balance errors and convergence rates. The following order in terms of decreasing efficiency is proposed: symmetric SI, Strang-splitting SNI, standard SNI, extrapolating SI and standard SI schemes. The symmetric SI scheme does not induce any operator-splitting errors, the Strang splitting SNI appears to be O(N2OS) accurate, and the other schemes are first order accurate. PMID- 14734249 TI - DNAPL remediation with in situ chemical oxidation using potassium permanganate. II. Increasing removal efficiency by dissolving Mn oxide precipitates. AB - In situ chemical oxidation (ISCO) schemes using MnO4- have been effective in destroying chlorinated organic solvents dissolved in ground water. Laboratory experiments and field pilot tests reveal that the precipitation of Mn oxide, one of the reaction products, causes a reduction of permeability, which can lead to flow bypassing and inefficiency of the scheme. Without a solution to this problem of plugging, it is difficult to remove DNAPL from the subsurface completely. In a companion paper, we showed with batch experiments that Mn oxide can be dissolved rapidly with certain organic acids. This study utilizes 2-D flow-tank experiments to examine the possibility of nearly complete DNAPL removal by ISCO with MnO4-, when organic acids are used to remove Mn oxide. The experiments were conducted in a small 2-D glass flow tank containing a lenticular silica-sand medium. Blue-dyed trichloroethylene (TCE) provided residual, the perched and pooled DNAPL. KMnO4 at 200 mg/l was flushed through the DNAPL horizontally. Once plugging reduced permeability and prevented further delivery of the oxidant, citric or oxalic acids were pumped into the flow tank to dissolve the Mn oxide precipitates. Organic ligands removed the Mn oxide precipitates relatively quickly, and permitted another cycle of MnO4- flooding. Cycles of MnO4-/acid flooding continued until all of the visible DNAPL was removed. The experiments were monitored with chemical analysis and visualization. A mass-balance calculation indicated that by the end of the experiments, all the DNAPL was removed. The results show also how heterogeneity adds complexity to initial redistribution of DNAPL, and to the efficiency of the chemical flooding. PMID- 14734250 TI - Probing metabolic pathways with isotopic tracers: insights from mammalian metabolic physiology. AB - Metabolic Engineering offers an opportunity to forge a link between metabolic physiologists, working with mammalian systems and metabolic engineers. Many parallels may be drawn between the specific modification of metabolic networks to improve cellular properties and the analysis of metabolic networks in search of causes of disease. At the core of both fields is the measurement of fluxes. This issue of Metabolic Engineering highlights important topics: mammalian metabolic physiology where estimating fluxes is challenging. The challenges come from compartmentation of metabolites, from dilution of tracer by endogenous pools, and from the difficulty of sampling relevant pools. The common theme across these investigations is the use of isotopic tracers. The wide variety of tracers and tracer analysis techniques in use in mammalian metabolic physiology reflects the complexity of the systems under study. In presenting these examples from the field of mammalian metabolic physiology, our goal is to strengthen the linkages between physiologists and engineers as we develop our knowledge and appreciation of the complexity of metabolic networks. PMID- 14734251 TI - A window into cellular metabolism: hepatic metabolism of (15)N-labelled substrates. AB - It is now apparent that many of the subtleties of cellular metabolism are intrinsically associated with cell structure and that their physiological study requires techniques that respect the integrity of cells and organs. We have used 15N-substrates to examine urea synthesis in the intact perfused rat liver. This work permits us to determine the extent to which different amino acids donate nitrogen atoms to the two nitrogens of urea. It is apparent that alanine and the amino group of glutamine provide nitrogen for urea synthesis primarily via cytoplasmic aspartate, whereas mitochondrial ammonia is the preferred route of entry for nitrogen from pre-formed ammonia or from the amide nitrogen of glutamine. Most importantly, this methodology permits us to explore for the occurrence of metabolic channels in such a highly organised, physiological system. Our studies indicate that a metabolic channel does not exist between glutaminase and carbamoylphosphate synthetase 1. PMID- 14734252 TI - Analytical solutions for (13)C isotopomer analysis of complex metabolic conditions: substrate oxidation, multiple pyruvate cycles, and gluconeogenesis. PMID- 14734253 TI - Using isotope tracers to study metabolism: application in mouse models. AB - The application of isotope tracers for investigating metabolism in mice is discussed. To familiarize the reader, some basic principles regarding the use of tracer methods are outlined. Emphasis is placed on showing how investigators are using isotope tracers to study the regulation of carbohydrate, fat and/or protein turnover in vivo. Finally, some of the advantages of using labeled water (i.e., 2H(2)O and/or H(2)18O) to trace the kinetics of biological processes are considered. The background provided in this report should assist engineers in designing studies that enhance our understanding of conditions in which metabolism is altered (e.g., diabetes, cancer cachexia, failure to thrive and travel at zero-gravity). PMID- 14734254 TI - Accumulation of 2-deoxy-D-glucose-6-phosphate as a measure of glucose uptake in the isolated perfused heart: a 31P NMR study. AB - The accumulation of 2-deoxy-D-glucose-6-phosphate (2DG6P), detected using 31P NMR spectroscopy, has been used as a measure of the rate of glucose uptake, yet the accuracy of this measurement has not been verified. In this study, isolated rat hearts were perfused with different substrates or isoproterenol for 30 min before measurement of either 2DG6P accumulation or [2-3H]glucose uptake, without and with insulin. Basal contractile function and metabolite concentrations were the same for all hearts. The basal rates of 2DG6P accumulation differed significantly, depending on the preceding perfusion protocol, and were 38-60% of the [2-3H]glucose uptake rates, whereas insulin-stimulated 2DG6P accumulation was the same or 71% higher than the [2-3H]glucose uptake rates. Therefore the ratio of 2DG6P accumulation/[2-3H]glucose uptake rates varied from 0.38 to 1.71, depending on the prior perfusion conditions or the presence of insulin. The rates of 2DG6P hydrolysis were found to be proportional to the intracellular 2DG6P concentrations, with a K(m) of 17.5mM and V(max) of 1.4 micromol/g dry weight/min. We conclude that the rates of 2DG6P accumulation do not accurately reflect glucose uptake rates under all physiological conditions in the isolated heart and should be used with caution. PMID- 14734255 TI - A critical perspective of the use of (13)C-isotopomer analysis by GCMS and NMR as applied to cardiac metabolism. AB - The aim of this article is to provide a guide for metabolic physiologists and bioengineers to the combined use of gas chromatography-mass spectrometry (GCMS) and nuclear magnetic resonance (NMR) in stable isotope investigations in any biological systems. Building on our past experience with these two techniques, as applied separately to the investigation of citric acid metabolism in the ex vivo perfused rat heart we initiated a collaborative study for their critical evaluation. This article, which expands on our previous work (Mol. Cel. Biol., 2003), directly compares GCMS- and NMR-determined 13C-isotopomer and flux data obtained from ex vivo rat heart perfusion studies with 13C-substrates. Overall we have found excellent agreement between the 13C-enrichments of GCMS- and NMR determined citric acid cycle metabolites (citrate, 2-ketoglutarate, succinate and malate) and glutamate; however the unlabeled component (M) was consistently underestimated by NMR. Despite this discrepancy there was reasonably good agreement in the relative fluxes of 13C-substrates through the citric acid cycle determined by the two techniques. Nevertheless, further investigations appear necessary before maximal advantage can be taken of the complementary 13C isotopomer and flux data of GCMS and NMR for probing the dynamics of cellular metabolism. PMID- 14734256 TI - Estimation of metabolic flux from dominant rate constants in vivo: application to brain and heart. AB - In an earlier paper (Cohen and Bergman, Am. J. Physiol. 268 (1995) E397), we explored the relationship between the exponents in the exponential curve fit to isotopic enrichment versus time and the fractional turnover rate of the largest metabolic pool in the pathway. Here we present the analysis on a more rigorous footing and apply it to questions of cerebral and cardiac metabolism. Our emphasis in this paper is to describe and justify mathematically an approach for analysis of metabolic dynamics, not with the intention of replacing the use of numerical software for estimation of flux rates but for giving the scientist the opportunity to examine the system in an approximate manner, and thereby to check not only that the results of the numerical solution are the correct solutions to the equations but also that the equations portray the correct simplification of the metabolic pathway. We introduce the "dominant rate constant" as a tool for deriving algebraic formulas relating rates of metabolic flux, sizes of metabolic pools, and the dynamics of isotopic enrichment. Illustrations of such algebraic formulas are provided for the rates of the citric acid cycle (CAC), glycolysis and glutamine synthesis in brain, as well as the rate of the CAC in heart. In addition, we prove that formulas for estimation of rates of glycolysis and of the CAC depend critically on the fractional turnover rates of lactate and glutamate, respectively. The justification for analysis of simulated data is that we are studying the effects of simplifications of metabolic models on the accuracy of estimation of metabolic pathways. Our use of the dominant rate constant is an analytical convenience that allows us to assess proposed simplifications of metabolic pathways. PMID- 14734257 TI - Basic principles of metabolic modeling of NMR (13)C isotopic turnover to determine rates of brain metabolism in vivo. AB - Metabolic modeling is a necessary part of the analysis of isotopic labeling data that is being obtained in the brain and other organs. Here are explained the basic principles of metabolic modeling of isotopic labeling studies, particularly with regard to (13)C isotopic measurements performed in vivo. The basic elements needed to simulate isotopic flows are described, and how to combine them to perform modeling analyses is explained. Procedures to introduce and evaluate model constraints and simplifications are discussed. The basic principle of isotopomer analysis is explained, as are mechanics of least-squares fitting of simulations to data. Closely related to the fitting is the effect of data scatter, which is discussed in the context of the non-normal distributions of uncertainty that are often seen with (13)C labeling measurements in vivo. This article is meant to provide a general background for investigators to begin to apply metabolic modeling analysis to (13)C isotopic labeling studies performed in vivo. PMID- 14734258 TI - New stable isotope-mass spectrometric techniques for measuring fluxes through intact metabolic pathways in mammalian systems: introduction of moving pictures into functional genomics and biochemical phenotyping. AB - The thesis of this review is that fully assembled metabolic pathways in living systems, rather than genes or proteins, are the true units of function in biology and biochemistry. A corollary is that measurement of metabolic fluxes (biochemical kinetics) is thereby required to understand biochemical control and gene function. Recent methodologic advances for improving observability of metabolic pathway fluxes in vivo are reviewed. Stable isotope-mass spectrometric techniques discussed here include mass isotopomer distribution analysis (combinatorial analysis), for measurement of polymerization biosynthesis; 2H(2)O administration, for measuring synthesis of DNA (i.e., cell proliferation), RNA, proteins, lipids, glycolipids and other classes of molecules; non-invasive probes of intracellular metabolism, by sampling secreted metabolites in accessible body fluids, after isotopic labeling of the intracellular pathway; and measurement of multiple molecular fluxes concurrently, particularly through use of 2H(2)O. Examples are given of pathway fluxes measured by each of these techniques, noting the often-surprising results. It is concluded that the introduction of "moving pictures" as tools for biochemical phenotyping could radically alter many signature areas of contemporary biology, including functional genomics, drug discovery and development, and disease research. PMID- 14734259 TI - Environmental conditions and variation in levels of sun exposure among children in child care. AB - Sun exposure in childhood is 1 of the risk factors for developing skin cancer, yet little is known about levels of exposure at this age. This is particularly important in countries with high levels of ultraviolet radiation (UVR) such as Australia. Among 49 children 3 to 5 years of age attending child care centers, UVR exposure was studied under 4 conditions in a repeated measures design; sunny days, cloudy days, teacher's instruction to stay in the shade, and a health professionals instruction to apply sunscreen. Three different data collection methods were employed: (a) completion of questionnaire or diary by parents and researcher, (b) polysulphone dosimeter readings, and (c) observational audits (video recording). Results of this study indicated that more than half the children had been sunburnt (pink or red) and more than a third had experienced painful sunburn (sore or tender) in the last summer. Most wore short sleeve shirts, short skirts or shorts and cap, that do not provide optimal levels of skin protection. However, sunscreen was applied to all exposed parts before the children went out to the playground. Over the period of 1 hr (9-10 a.m.) the average amount of time children spent in full sun was 22 min. On sunny days there was more variation across children in the amount of sun exposure received. While the potential amount of UVR exposure for young children during the hour they were outside on a sunny day was 1.45 MED (Minimum Erythemal Dose), they received on average 0.35 MED, which is an insufficient amount to result in an erythemal response on fair skin even without the use of sunscreen. PMID- 14734260 TI - Exploring the development of sun-tanning behavior: a grounded theory study of adolescents' decision-making experiences with becoming a sun tanner. AB - A grounded theory study was undertaken to describe how adolescents make decisions about sunbathing during the transition from childhood to adolescence and to propose an explanation for the relationships among factors affecting the adoption of sun tanning. In-depth interviews (n = 40) were conducted separately with adolescents (aged 12 to 16 years) and their parents. Constant comparative analysis of adolescents' accounts identified two methods that adolescents described as a means of getting a suntan: intentional sun tanning and incidental sun tanning. The process of adolescents' decision-making about getting a suntan can be understood by examining the following sequence: becoming motivated to get a tan, experimenting with sun tanning, and establishing self as an intentional tanner or incidental tanner. Implications for developing strategies to prevent the adoption of sun-tanning habits among adolescents are presented. PMID- 14734261 TI - Cholesterol changes in coronary patients after a short behavior modification program. AB - Serum cholesterol changes after an 8-week behavior modification program for patients with coronary artery disease (CAD) were studied in a randomized controlled clinical trial. Acute myocardial infarction (AMI) or coronary artery bypass grafting (CABG) patients were randomly assigned to the intervention (N = 94) or to usual care (N = 90). After 9 months' follow-up the intervention was effective in reducing total cholesterol and LDL cholesterol levels, particularly in patients with high baseline lipid levels. After correcting for changes in dose of statins during follow-up, effects were weakened, but for patients with high baseline cholesterol levels favorable effects remained. In these patients, the intervention group showed a decline of total cholesterol and LDL cholesterol levels of 20% and 29%, respectively, compared to a 12% and 19% reduction in the control group (p <.01). These effects could not be explained by changes in dietary fat consumption. An unexpected finding was a lower increase in HDL cholesterol in the intervention group than in the control group. PMID- 14734262 TI - Coping strategies, hostility, and depressive symptoms: a path model. AB - Previous studies of coping, hostility, and depressive symptoms have highlighted the significant relations between all possible pairs of these 3 variables. To more completely explore the nature of depressive symptoms, we link them all together in this study by testing a coping-->hostility-->depressive symptoms path model. One hundred forty participants completed psychological questionnaires measuring coping strategies, hostility, and depressive symptoms. While controlling age and social class as covariates, SPSS stepwise regression analyses were used to examine relations among these 3 constructs. Results suggest that coping has a direct relation with depressive symptoms as well as an indirect relation mediated by hostility. Passive coping may lead to increased hostility, resulting in depressive symptoms. Active coping may have the opposite effect. These findings suggest that the inclusion of measures of both coping strategies and hostility yields a more thorough understanding of concomitants of depressive symptoms. From a clinical perspective, knowing what coping strategies a person uses and how much anger they experience and express may be useful in guiding the management of depressive symptoms. PMID- 14734263 TI - Pain-coping strategies in chronic pain patients: psychometric characteristics of the pain-coping inventory (PCI). AB - This article presents a series of studies aimed at validating a comprehensive pain-coping inventory (PCI) that is applicable to various types of patients with chronic pain. Item and scale analyses were performed for patients with rheumatoid arthritis (RA), patients with chronic headache, and pain clinic outpatients. The following 6 scales were derived from a simultaneous component analysis: Pain Transformation, Distraction, Reducing Demands, Retreating, Worrying, and Resting, all of which were internally reliable. A higher order factor analysis grouped the PCI scales into active (transformation, distraction, reducing demands) and passive (retreating, worrying, resting) pain-coping dimensions. Differences in use of strategy found between RA patients and headache patients indicated that the PCI scales were sufficiently sensitive to measure differences between groups. Concurrent validity was assessed for patients with RA and patients with fibromyalgia and predictive validity was assessed for patients with recently diagnosed RA after 1 and 3 years. In both analyses the validity of the scales was supported, in particular the predictive validity of passive coping scales for future outcomes. PMID- 14734264 TI - Phenotypic and genotypic characteristics of women in relation to personality traits. AB - The associations were examined in women between personality traits and steroid hormones, particularly androgens, as well as polymorphisms in genes regulating androgen concentration and effects. Women, all 42 years of age and premenopausal (n = 270), were recruited randomly. Conventional "masculine" and "feminine" personality traits were examined by questionnaire and set in relation to psychosocial and socioeconomic conditions, behavior in childhood, hormones, risk factors for disease, and polymorphisms in microsatellites in the CYP aromatase and the androgen receptor gene. The proportions of personality traits considered as being dominated by "masculinity" (M) or "femininity" (F) were 44.9%, respectively 15.0%, the rest consisting of a combination of M and F (33.2%) or "undifferentiated" (6.9%). M characteristics were positively associated with education, sporting, self-confidence, and good adaptation to work situation. M scores correlated with reports of "tomboyism" as girls. There was essentially no difference in hormones or disease risk factors between M and F women. The number of (CAG) repeats in the microsatellite of the transactivating domain of the androgen receptor was 19 (2.3; M and SD). M characteristics were more pronounced in the presence of longer repeat stretches (n > 20). No associations were found with F scores. There were no significant associations to the number of tetranucleotide repeats (TTTA) in the fourth introne of the aromatase gene. It was concluded that a majority of women showed M type of personality traits, associated with normal hormones, somatic health, and a long microsatellite in the transactivating domain of the AR gene. PMID- 14734266 TI - Four for 2004: how to become physically active. We know that physical activity is good for us, but how do we get going? PMID- 14734267 TI - Update on black cohosh for hot flashes. PMID- 14734268 TI - The best way to treat fibromyalgia. It may require more than one strategy, but you can get some pain relief and feel a lot better about life. PMID- 14734269 TI - Living wills and health care proxies. Documenting your health care wishes can help ensure that your voice will be heard should you be unable to speak for yourself. PMID- 14734270 TI - By the way, doctor. I'm 60 years old and have always been told my blood pressure, which is around 130/80, is normal. According to recent medical news, I have a condition called "prehypertension". Do I have high blood pressure or not? And should I be treated for it? PMID- 14734271 TI - By the way, doctor. My doctor told me I'm overdue for a tetanus booster. I'm 60, and I wonder if it's really necessary at my age. Can't I just get one if I get a deep cut? PMID- 14734272 TI - High blood pressure: treatment guidelines. How you can silence the "silent killer". PMID- 14734273 TI - An update on calcium and prostate cancer. PMID- 14734274 TI - Diet and cholesterol: why bother? PMID- 14734275 TI - Hot tub sense and safety. PMID- 14734276 TI - Pedophilia. Who are the men who "love" children in intolerable ways? And how can they be helped to change? PMID- 14734277 TI - Pharmacogenomics in psychiatry. PMID- 14734278 TI - SAMe reconsidered. PMID- 14734279 TI - Bipolar disorder and schizophrenia: a common basis? PMID- 14734280 TI - Life-giving support. PMID- 14734281 TI - Questions & answers. Can someone suffer from a "shy" bladder? What is the proper term to use when a person can't urinate in front of someone else, as when giving a sample for a drug test? What causes the problem, and can it be treated? PMID- 14734282 TI - Should you enroll in a clinical trial? Volunteering for a study is a great public service, but don't forget that you're participating in an experiment. Know the risks, and ask plenty of questions. PMID- 14734283 TI - Are they better than Viagra? Two new drugs for erectile dysfunction work like Viagra and carry similar risks and benefits. Their subtle differences, however, may make a difference for some men. PMID- 14734284 TI - Cancer: how not to go there. The secret to cancer prevention may be in the gym. Or in the spice rack. Let's hope researchers find it soon. PMID- 14734285 TI - Dietary fat. PMID- 14734286 TI - Six steps to help your knees. PMID- 14734287 TI - Antibiotics: Achilles' heel of the Achilles' tendon. PMID- 14734288 TI - Sweet stuff may sour your digestion. PMID- 14734289 TI - By the way, doctor. Much is made about high blood pressure, but little is said when it's really low. At what levels are systolic and diastolic readings considered too low? Are there any symptoms? What about treatments? PMID- 14734290 TI - By the way, doctor. Why do doctors try to determine if a person has heart disease by asking about smoking, high cholesterol, or family history? Why can't they just examine the heart and blood vessels directly, and get a precise answer? PMID- 14734291 TI - Planning pays off if a heart attack strikes. Knowing the warning signs and what to do if they appear can mean the difference between rapid recovery and long-term heart damage. PMID- 14734292 TI - Artery opening trumps clot busting. Angioplasty for heart attacks may prompt a shift to regional "heart attack centers". PMID- 14734293 TI - Drug-coated stents boost benefits of angioplasty. These tiny devices keep unclogged arteries open but don't prevent blockages from arising in other heart arteries. PMID- 14734294 TI - Getting the most from cardiac rehabilitation. Almost anyone with heart disease can benefit from a comprehensive program of exercise and education. PMID- 14734295 TI - The shape of healthy eating. Confused by what constitutes healthy eating? Here's a quick guide based on the latest and best evidence. PMID- 14734296 TI - Underestimating the hazards of peripheral artery disease. PMID- 14734298 TI - Age no limit for bypass surgery. PMID- 14734297 TI - Shovel as if your heart mattered. PMID- 14734299 TI - Hidden angina. PMID- 14734300 TI - Targeting HDL. PMID- 14734301 TI - Ask the doctor. Someone forwarded an e-mail to me about coughing during a heart attack. It said that if I am having a heart attack, coughing deeply and strongly every few seconds could save my life. Is that true? PMID- 14734303 TI - A strategy for assembling the maize (Zea mays L.) genome. AB - Because the bulk of the maize (Zea mays L.) genome consists of repetitive sequences, sequencing efforts are being targeted to its 'gene-rich' fraction. Traditional assembly programs are inadequate for this approach because they are optimized for a uniform sampling of the genome and inherently lack the ability to differentiate highly similar paralogs. RESULTS: We report the development of bioinformatics tools for the accurate assembly of the maize genome. This software, which is based on innovative parallel algorithms to ensure scalability, assembled 730,974 genomic survey sequences fragments in 4 h using 64 Pentium III 1.26 GHz processors of a commodity cluster. Algorithmic innovations are used to reduce the number of pairwise alignments significantly without sacrificing quality. Clone pair information was used to estimate the error rate for improved differentiation of polymorphisms versus sequencing errors. The assembly was also used to evaluate the effectiveness of various filtering strategies and thereby provide information that can be used to focus subsequent sequencing efforts. PMID- 14734304 TI - Multi-species comparative mapping in silico using the COMPASS strategy. AB - MOTIVATION: The completion of human and mouse genome sequences provides a valuable resource for decoding other mammalian genomes. The comparative mapping by annotation and sequence similarity (COMPASS) strategy takes advantage of the resource and has been used in several genome-mapping projects. It uses existing comparative genome maps based on conserved regions to predict map locations of a sequence. An automated multiple-species COMPASS tool can facilitate in the genome sequencing effort and comparative genomics study of other mammalian species. RESULTS: The prerequisite of COMPASS is a comparative map table between the reference genome and the predicting genome. We have built and collected comparative maps among five species including human, cattle, pig, mouse and rat. Cattle-human and pig-human comparative maps were built based on the positions of orthologous markers and the conserved synteny groups between human and cattle and human and pig genomes, respectively. Mouse-human and rat-human comparative maps were based on the conserved sequence segments between the two genomes. With a match to human genome sequences, the approximate location of a query sequence can be predicted in cattle, pig, mouse and rat genomes based on the position of the match relatively to the orthologous markers or the conserved segments. AVAILABILITY: The COMPASS-tool and databases are available at http://titan.biotec.uiuc.edu/COMPASS/ PMID- 14734305 TI - Transposable element annotation of the rice genome. AB - MOTIVATION: The high content of repetitive sequences in the genomes of many higher eukaryotes renders the task of annotating them computationally intensive. Presently, the only widely accepted method of searching and annotating transposable elements (TEs) in large genomic sequences is the use of the RepeatMasker program, which identifies new copies of TEs by pairwise sequence comparisons with a library of known TEs. Profile hidden Markov models (HMMs) have been used successfully in discovering distant homologs of known proteins in large protein databases, but this approach has only rarely been applied to known model TE families in genomic DNA. RESULTS: We used a combination of computational approaches to annotate the TEs in the finished genome of Oryza sativa ssp. japonica. In this paper, we discuss the strengths and the weaknesses of the annotation methods used. These approaches included: the default configuration of RepeatMasker using cross_match, an implementation of the Smith-Waterman-Gotoh algorithm; RepeatMasker using WU-BLAST for similarity searching; and the HMMER package, used to search for TEs with profile HMMs. All the results were converted into GFF format and post-processed using a set of Perl scripts. RepeatMasker was used in the case of most TE families. The WU-BLAST implementation of RepeatMasker was found to be manifold faster than cross_match with only a slight loss in sensitivity and was thus used to obtain the final set of data. HMMER was used in the annotation of the Mutator-like element (MULE) superfamily and the miniature inverted-repeat transposable element (MITE) polyphyletic group of families, for which large libraries of elements were available and which could be divided into well-defined families. The HMMER search algorithm was extremely slow for models over 1000 bp in length, so MULE families with members over 1000 bp long were processed with RepeatMasker instead. The main disadvantage of HMMER in this application is that, since it was developed with protein sequences in mind, it does not search the negative DNA strand. With the exception of TE families with essentially palindromic sequences, reverse complement models had to be created and run to compensate for this shortcoming. We conclude that a modification of RepeatMasker to incorporate libraries of profile HMMs in searches could improve the ability to detect degenerated copies of TEs. AVAILABILITY: The Perl scripts and TE sequences used in construction of the RepeatMasker library and the profile HMMs are available upon request. PMID- 14734306 TI - Computational identification of novel chitinase-like proteins in the Drosophila melanogaster genome. AB - MOTIVATION: Multiple chitinases as well as lectins closely related to them have been characterized previously from many insect species and the corresponding genes/cDNAs have been cloned. However, the identification of the entire assortment of genes for chitinase family proteins and their differences in biochemical properties have not been carried out in any individual insect species. The completion of the entire DNA sequence of Drosophila melanogaster (fruit fly) genome and identification of open reading frames presents an opportunity to study the structures and functions of chitinase-like proteins, and also to identify new members of this family in Drosophila. We are, therefore, interested in studying the functional genomics of chitinase-like gene families in insects. METHODS: We searched the Drosophila protein sequences database using fully characterized insect chitinase sequences and BLASTP software, identified all the putative chitinase-like proteins encoded in Drosophila genome, and predicted their structures using domain analysis tools. A phylogenetic analysis of the chitinase-like proteins from Drosophila and several other insect species was carried out. The structures of these chitinases were modeled using homology modeling software. RESULTS: Our analysis revealed the presence of 18 chitinase like proteins in the Drosophila protein database. Among these are seven novel chitinase-like proteins that contain four signature amino acid sequences of chitinases belonging to family 18 glycosylhydrolases, including both acidic and hydrophobic amino acid residues critical for enzyme activity. All the proteins contain at least one catalytic domain with one having four catalytic domains. Phylogenetic analysis of chitinase-like proteins from Drosophila and other insects revealed an evolutionary relationship among all these proteins, which indicated gene duplication and domain shuffling to generate the observed diversity in the encoded proteins. Homology modeling showed that all the Drosophila chitinase-like proteins contain one or more catalytic domains with a (alpha/beta)8 barrel-like structure. Our results suggest that insects utilize multiple family 18 chitinolytic enzymes and also non-enzymatic chitinase-like proteins for degrading/remodeling/binding to chitin in different insect anatomical extracellular structures, such as the cuticle, peritrophic membrane, trachea and mouth parts during insect development, and possibly for other roles including chitin synthesis. AVAILABILITY: Perl program and supplementary material are available at http://www.ksu.edu/bioinformatics/supplementary.htm PMID- 14734307 TI - Phylogenomic inference of protein molecular function: advances and challenges. AB - MOTIVATION: Protein families evolve a multiplicity of functions through gene duplication, speciation and other processes. As a number of studies have shown, standard methods of protein function prediction produce systematic errors on these data. Phylogenomic analysis--combining phylogenetic tree construction, integration of experimental data and differentiation of orthologs and paralogs- has been proposed to address these errors and improve the accuracy of functional classification. The explicit integration of structure prediction and analysis in this framework, which we call structural phylogenomics, provides additional insights into protein superfamily evolution. RESULTS: Results of protein functional classification using phylogenomic analysis show fewer expected false positives overall than when pairwise methods of functional classification are employed. We present an overview of the motivations and fundamental principles of phylogenomic analysis, new methods developed for the key tasks, benchmark datasets for these tasks (when available) and suggest procedures to increase accuracy. We also discuss some of the methods used in the Celera Genomics high throughput phylogenomic classification of the human genome. AVAILABILITY: Software tools from the Berkeley Phylogenomics Group are available at http://phylogenomics.berkeley.edu PMID- 14734308 TI - A comparative phylogenetic approach for dating whole genome duplication events. AB - MOTIVATION: Whole genome duplications have played a major role in determining the structure of eukaryotic genomes. Current evidence revealing large blocks of duplicated chromatin yields new insights into the evolutionary history of species, but also presents a major challenge for researchers attempting to utilize comparative genomics techniques. Understanding the timing of duplication events relative to divergence among taxa is critical to accurate and comprehensive cross-species comparisons. RESULTS: We describe a large-scale approach to estimate the timing of duplication events in a phylogenetic context. The methodology has been previously utilized for analysis of Arabidopsis and Saccharomyces duplication events. This new implementation provides a more flexible and reusable framework for these analyses. Scripts written in the Python programming language drive a number of freely available bioinformatics programs, creating a no-cost tool for researchers. The usefulness of the approach is demonstrated through genome-scale analysis of Arabidopsis and Oryza (rice) duplications. AVAILABILITY: Software and documentation are freely available from http://plantgenome.agtec.uga.edu/bioinformatics/dating/ PMID- 14734309 TI - Prediction of locally stable RNA secondary structures for genome-wide surveys. AB - MOTIVATION: Recently novel classes of functional RNAs, most prominently the miRNAs have been discovered, strongly suggesting that further types of functional RNAs are still hidden in the recently completed genomic DNA sequences. Only few techniques are known, however, to survey genomes for such RNA genes. When sufficiently similar sequences are not available for comparative approaches the only known remedy is to search directly for structural features. RESULTS: We present here efficient algorithms for computing locally stable RNA structures at genome-wide scales. Both the minimum energy structure and the complete matrix of base pairing probabilities can be computed in theta(N x L2) time and theta(N + L2) memory in terms of the length N of the genome and the size L of the largest secondary structure motifs of interest. In practice, the 100 Mb of the complete genome of Caenorhabditis elegans can be folded within about half a day on a modern PC with a search depth of L = 100. This is sufficient example for a survey for miRNAs. AVAILABILITY: The software described in this contribution will be available for download at http://www.tbi.univie.ac.at/~ivo/RNA/ as part of the Vienna RNA Package. PMID- 14734310 TI - Shared relationship analysis: ranking set cohesion and commonalities within a literature-derived relationship network. AB - MOTIVATION: There is a general scientific need to be able to identify and evaluate what any given set of 'objects' (e.g. genes, phenotypes, chemicals, diseases) has in common. Whether it is to classify, expand upon or identify commonalities and functional groupings, informational needs can be diverse and the best source to identify relationships among a potentially heterogeneous set of objects is the scientific literature. RESULTS: We first establish a network of related objects by their co-occurrence within MEDLINE records. A set of objects within this network can then be queried to identify shared relationships, and a method is presented to score their statistical relevance by comparing observed frequencies with what would be expected in a random network model. Using Gene Ontology (GO) categories, we demonstrate that this method enables a quantitative ranking of the 'cohesiveness' of a set of objects and, importantly, allows other objects related to this set to be identified and evaluated for their 'cohesion' to it. Supplemental information: A list of ranked genes related to each GO category analyzed can be found at http://innovation.swmed.edu/IRIDESCENT/GO_relationships.htm PMID- 14734311 TI - Limited conformational space for early-stage protein folding simulation. AB - MOTIVATION: The problem of early-stage protein folding is critical for protein structure prediction. The model presented introduces a common definition of protein structures which may be treated as the possible in silico early-stage form of the polypeptide chain. Limitation of the conformational space to the ellipse path on the Ramachandran map was tested as a possible sub-space to represent the early-stage structure for simulation of protein folding. The proposed conformational sub-space was developed on the basis of the backbone conformation, with side-chain interactions excluded. RESULTS: The ellipse-path limited conformation of BPTI was created using the criterion of shortest distance between Phi, Psi angles in native form of protein and the Phi, Psi angles belonging to the ellipse. No knots were observed in the structure created according to ellipse-path conformational sub-space. The energy minimization procedure applied to ellipse-path derived conformation directed structural changes toward the native form of the protein with SS-bonds system introduced to the procedure. AVAILABILITY: Program 'Ellipse' to create the ellipse-path derived structure available on request: myroterm@cyf-kr.edu.pl PMID- 14734312 TI - Comparative evaluation of word composition distances for the recognition of SCOP relationships. AB - MOTIVATION: Alignment-free metrics were recently reviewed by the authors, but have not until now been object of a comparative study. This paper compares the classification accuracy of word composition metrics therein reviewed. It also presents a new distance definition between protein sequences, the W-metric, which bridges between alignment metrics, such as scores produced by the Smith-Waterman algorithm, and methods based solely in L-tuple composition, such as Euclidean distance and Information content. RESULTS: The comparative study reported here used the SCOP/ASTRAL protein structure hierarchical database and accessed the discriminant value of alternative sequence dissimilarity measures by calculating areas under the Receiver Operating Characteristic curves. Although alignment methods resulted in very good classification accuracy at family and superfamily levels, alignment-free distances, in particular Standard Euclidean Distance, are as good as alignment algorithms when sequence similarity is smaller, such as for recognition of fold or class relationships. This observation justifies its advantageous use to pre-filter homologous proteins since word statistics techniques are computed much faster than the alignment methods. AVAILABILITY: All MATLAB code used to generate the data is available upon request to the authors. Additional material available at http://bioinformatics.musc.edu/wmetric PMID- 14734313 TI - GAPSCORE: finding gene and protein names one word at a time. AB - MOTIVATION: New high-throughput technologies have accelerated the accumulation of knowledge about genes and proteins. However, much knowledge is still stored as written natural language text. Therefore, we have developed a new method, GAPSCORE, to identify gene and protein names in text. GAPSCORE scores words based on a statistical model of gene names that quantifies their appearance, morphology and context. RESULTS: We evaluated GAPSCORE against the Yapex data set and achieved an F-score of 82.5% (83.3% recall, 81.5% precision) for partial matches and 57.6% (58.5% recall, 56.7% precision) for exact matches. Since the method is statistical, users can choose score cutoffs that adjust the performance according to their needs. AVAILABILITY: GAPSCORE is available at http://bionlp.stanford.edu/gapscore/ PMID- 14734314 TI - Minimal cut sets in biochemical reaction networks. AB - MOTIVATION: Structural studies of metabolic networks yield deeper insight into topology, functionality and capabilities of the metabolisms of different organisms. Here, we address the analysis of potential failure modes in metabolic networks whose occurrence will render the network structurally incapable of performing certain functions. Such studies will help to identify crucial parts in the network structure and to find suitable targets for repressing undesired metabolic functions. RESULTS: We introduce the concept of minimal cut sets for biochemical networks. A minimal cut set (MCS) is a minimal (irreducible) set of reactions in the network whose inactivation will definitely lead to a failure in certain network functions. We present an algorithm which enables the computation of the MCSs in a given network related to user-defined objective reactions. This algorithm operates on elementary modes. A number of potential applications are outlined, including network verifications, phenotype predictions, assessing structural robustness and fragility, metabolic flux analysis and target identification in drug discovery. Applications are illustrated by the MCSs in the central metabolism of Escherichia coli for growth on different substrates. AVAILABILITY: Computation and analysis of MCSs is an additional feature of the FluxAnalyzer (freely available for academic users upon request, special contracts for industrial companies; see web page below). SUPPLEMENTARY INFORMATION: http://www.mpi-magdeburg.mpg.de/projects/fluxanalyzer PMID- 14734315 TI - An empirical bayes adjustment to increase the sensitivity of detecting differentially expressed genes in microarray experiments. AB - MOTIVATION: Detection of differentially expressed genes is one of the major goals of microarray experiments. Pairwise comparison for each gene is not appropriate without controlling the overall (experimentwise) type 1 error rate. Dudoit et al. have advocated use of permutation-based step-down P-value adjustments to correct the observed significance levels for the individual (i.e. for each gene) two sample t-tests. RESULTS: In this paper, we consider an ANOVA formulation of the gene expression levels corresponding to multiple tissue types. We provide resampling-based step-down adjustments to correct the observed significance levels for the individual ANOVA t-tests for each gene and for each pair of tissue type comparisons. More importantly, we introduce a novel empirical Bayes adjustment to the t-test statistics that can be incorporated into the step-down procedure. Using simulated data, we show that the empirical Bayes adjustment improved the sensitivity of detecting differentially expressed genes up to 16%, while maintaining a high level of specificity. This adjustment also reduces the false non-discovery rate to some degree at the cost of a modest increase in the false discovery rate. We illustrate our approach using a human colon cancer dataset consisting of oligonucleotide arrays of normal, adenoma and carcinoma cells. The number of genes with differential expression level declared statistically significant was about 50 when comparing normal to adenoma cells and about five when comparing adenoma to carcinoma cells. This list includes genes previously known to be associated with colon cancer as well as some novel genes. AVAILABILITY: R code for the empirical Bayes adjustment and step-down P-value calculation via resampling are available from the supplementary web-site. SUPPLEMENTARY INFORMATION: http://www.mathstat.gsu.edu/~matsnd/EB/supp.htm PMID- 14734316 TI - Graph-based clustering for finding distant relationships in a large set of protein sequences. AB - MOTIVATION: Clustering of protein sequences is widely used for the functional characterization of proteins. However, it is still not easy to cluster distantly related proteins, which have only regional similarity among their sequences. It is therefore necessary to develop an algorithm for clustering such distantly related proteins. RESULTS: We have developed a time and space efficient clustering algorithm. It uses a graph representation where its vertices and edges denote proteins and their sequence similarities above a certain cutoff score, respectively. It repeatedly partitions the graph by removing edges that have small weights, which correspond to low sequence similarities. To find the appropriate partitions, we introduce a score combining the normalized cut and a locally minimal cut capacities. Our method is applied to the entire 40,703 human proteins in SWISS-PROT and TrEMBL. The resulting clusters shows a 76% recall (20,529 proteins) of the 26,917 classified by InterPro. It also finds relationships not found by other clustering methods. AVAILABILITY: The complete result of our algorithm for all the human proteins in SWISS-PROT and TrEMBL, and other supplementary information are available at http://motif.ics.es.osaka u.ac.jp/Ncut-KL/ PMID- 14734317 TI - Is cross-validation better than resubstitution for ranking genes? AB - MOTIVATION: Ranking gene feature sets is a key issue for both phenotype classification, for instance, tumor classification in a DNA microarray experiment, and prediction in the context of genetic regulatory networks. Two broad methods are available to estimate the error (misclassification rate) of a classifier. Resubstitution fits a single classifier to the data, and applies this classifier in turn to each data observation. Cross-validation (in leave-one-out form) removes each observation in turn, constructs the classifier, and then computes whether this leave-one-out classifier correctly classifies the deleted observation. Resubstitution typically underestimates classifier error, severely so in many cases. Cross-validation has the advantage of producing an effectively unbiased error estimate, but the estimate is highly variable. In many applications it is not the misclassification rate per se that is of interest, but rather the construction of gene sets that have the potential to classify or predict. Hence, one needs to rank feature sets based on their performance. RESULTS: A model-based approach is used to compare the ranking performances of resubstitution and cross-validation for classification based on real-valued feature sets and for prediction in the context of probabilistic Boolean networks (PBNs). For classification, a Gaussian model is considered, along with classification via linear discriminant analysis and the 3-nearest-neighbor classification rule. Prediction is examined in the steady-distribution of a PBN. Three metrics are proposed to compare feature-set ranking based on error estimation with ranking based on the true error, which is known owing to the model-based approach. In all cases, resubstitution is competitive with cross validation relative to ranking accuracy. This is in addition to the enormous savings in computation time afforded by resubstitution. PMID- 14734318 TI - Formalization of mouse embryo anatomy. AB - MOTIVATION: The Edinburgh Mouse Atlas and Gene Expression Database project has developed a digital atlas of mouse development to provide a spatio-temporal framework for spatially mapped data such as in situ gene expression and cell lineage. As part of this database, a mouse embryo anatomy ontology has been created. A formalization of this anatomy is required to document its precise semantics and how it is used in the context of the Mouse Atlas. RESULTS: The paper describes the existing anatomy ontology and formalizes aspects of it using a predicate logic based approach. It therefore provides a guide for users of the current version of the ontology, as well as the basis for a description of the anatomy using an ontology language, such as OWL, thus enabling future work on reasoning about the Mouse Atlas in the context of an intelligent gene expression bioinformatics workflow system. The logic has been implemented in a Prolog prototype. AVAILABILITY: The Mouse Atlas is available on-line at http://genex.hgu.mrc.ac.uk PMID- 14734319 TI - The Helmholtz Network for Bioinformatics: an integrative web portal for bioinformatics resources. AB - SUMMARY: The Helmholtz Network for Bioinformatics (HNB) is a joint venture of eleven German bioinformatics research groups that offers convenient access to numerous bioinformatics resources through a single web portal. The 'Guided Solution Finder' which is available through the HNB portal helps users to locate the appropriate resources to answer their queries by employing a detailed, tree like questionnaire. Furthermore, automated complex tool cascades ('tasks'), involving resources located on different servers, have been implemented, allowing users to perform comprehensive data analyses without the requirement of further manual intervention for data transfer and re-formatting. Currently, automated cascades for the analysis of regulatory DNA segments as well as for the prediction of protein functional properties are provided. AVAILABILITY: The HNB portal is available at http://www.hnbioinfo.de PMID- 14734320 TI - ROSO: optimizing oligonucleotide probes for microarrays. AB - ROSO is software to design optimal oligonucleotide probe sets for microarrays. Selected probes show no significant cross-hybridization, no stable secondary structures and their Tm are chosen to minimize the Tm variability of the probe set. AVAILABILITY: The program is available on the internet. Sources are freely available, for non-profit use, on request to the authors. SUPPLEMENTARY INFORMATION: http://pbil.univ-lyon1.fr/roso PMID- 14734321 TI - LVB: parsimony and simulated annealing in the search for phylogenetic trees. AB - The program LVB seeks parsimonious phylogenies from nucleotide alignments, using the simulated annealing heuristic. LVB runs fast and gives high quality results. AVAILABILITY: The software is available at http://www.rubic.reading.ac.uk/lvb/ SUPPLEMENTARY INFORMATION: SUPPLEMENTARY INFORMATION may be downloaded from http://www.rubic.reading.ac.uk/~daniel/ PMID- 14734322 TI - PGTdb: a database providing growth temperatures of prokaryotes. AB - Included in Prokaryotic Growth Temperature database (PGTdb) are a total of 1334 temperature data from 1072 prokaryotic organisms, Bacteria and Archaea: PGTdb integrates microbial growth temperature data from literature survey with their nucleotide/protein sequence and protein structure data from related databases. A direct correlation is observed between the average growth temperature of an organism and the melting temperature of proteins from the organism. Therefore, this database is useful not only for microbiologists to obtain cultivation condition, but also for biochemists and structure biologists to study the correlation between protein sequences/structures and their thermostability. In addition, the taxonomy and ribosomal RNA sequence(s) of an organism are linked through NCBI Taxonomy and the Ribosomal RNA Operon Copy Number Database umdb, respectively. PGTdb is the only integrated database on the Internet to provide the growth temperature data of the prokaryotes and the combined information of their nucleotide/protein sequences, protein structures, taxonomy and phylogeny. AVAILABILITY: http://pgtdb.csie.ncu.edu.tw PMID- 14734323 TI - JDotter: a Java interface to multiple dotplots generated by dotter. AB - Java-Dotter (JDotter) is a platform-independent Java interactive interface for the Linux version of Dotter, a widely used program for generating dotplots of large DNA or protein sequences. JDotter runs as a client-server application and can send new sequences to the Dotter program for alignment as well as rapidly access a repository of preprocessed dotplots. JDotter also interfaces with a sequence database or file system to display supplementary feature data. Thus, JDotter greatly simplifies access to dotplot data in laboratories that deal with large numbers of genomes and have a multi-platform organization. AVAILABILITY: Currently, JDotter is used via Java Web Start by the Poxvirus Bioinformatics Resource for examining dotplots of complete poxvirus genomes; http://athena.bioc.uvic.ca/pbr/jdotter/. The software is available for download from the same location. SUPPLEMENTARY INFORMATION: Installation instructions, the User's Manual, screenshots and examples are available at the JDotter home page http://athena.bioc.uvic.ca/pbr/jdotter/. The software and source code is free for non-commercial applications. PMID- 14734324 TI - FGDP: functional genomics data pipeline for automated, multiple microarray data analyses. AB - Gene expression microarrays and oligonucleotide GeneChips have provided biologists with a means of measuring, in a single experiment, the expression levels of entire genomes under a variety of conditions. As with any nascent field, there is no single accepted method for analyzing the new data types, with new methods appearing monthly. Investigators using the new technology must constantly seek access to the latest tools and explore their data in multiple ways. The functional genomics data pipeline provides an integrated, extendable analysis environment permitting multiple, simultaneous analyses to be automatically performed and provides a web server and interface for presenting results. AVAILABILITY: Source code and executables are available under the GNU public license at http://bioinformatics.fccc.edu/ PMID- 14734325 TI - YETI: Yeast Exploration Tool Integrator. AB - Yeast Exploration Tool Integrator (YETI) is a novel bioinformatics tool for the integrated visualization and analysis of functional genomic data sets from the budding yeast Saccharomyces cerevisiae. AVAILABILITY: YETI is freely available for use over the WWW, or download under license, at http://www.bru.ed.ac.uk/~orton/yeti.html PMID- 14734326 TI - EST-PAGE--managing and analyzing EST data. AB - EST-PAGE provides a bioinformatics solution for expressed sequence tags (EST) data entry, database management, GenBank submission, process control and data retrieval from a unified web interface that can be easily customized and adapted by groups working on diverse EST sequencing projects. AVAILABILITY: The system and source code are available upon request from the authors. SUPPLEMENTARY INFORMATION: http://EST-PAGE.binf.gmu.edu PMID- 14734327 TI - APE: Analyses of Phylogenetics and Evolution in R language. AB - Analysis of Phylogenetics and Evolution (APE) is a package written in the R language for use in molecular evolution and phylogenetics. APE provides both utility functions for reading and writing data and manipulating phylogenetic trees, as well as several advanced methods for phylogenetic and evolutionary analysis (e.g. comparative and population genetic methods). APE takes advantage of the many R functions for statistics and graphics, and also provides a flexible framework for developing and implementing further statistical methods for the analysis of evolutionary processes. AVAILABILITY: The program is free and available from the official R package archive at http://cran.r project.org/src/contrib/PACKAGES.html#ape. APE is licensed under the GNU General Public License. PMID- 14734328 TI - AnaGram: protein function assignment. AB - SUMMARY: AnaGram is a web service for protein function assignment based on identity detection of small significant fragments (protomotifs) that can act as modular pieces in peptide construction. The system is able to assign function by finding correlations between protomotifs and functional annotations contained in SWISS-PROT and Medline databases. In addition, function ontologies are used for hierarchical organization of the predicted functions. Extensive tests have been carried out to evaluate the accuracy and performance of the system. AVAILABILITY: http://jaguar.genetica.uma.es/anagram.htm PMID- 14734329 TI - Graded activity for low back pain in occupational health care: a randomized, controlled trial. AB - BACKGROUND: Low back pain is a common medical and social problem frequently associated with disability and absence from work. However, data on effective return to work after interventions for low back pain are scarce. OBJECTIVE: To determine the effectiveness of a behavior-oriented graded activity program compared with usual care. DESIGN: Randomized, controlled trial. SETTING: Occupational health services department of an airline company in the Netherlands. PATIENTS: 134 workers who were absent from work because of low back pain were randomly assigned to either graded activity (n = 67) or usual care (n = 67). INTERVENTION: Graded activity, a physical exercise program based on operant conditioning behavioral principles, to stimulate a rapid return to work. MEASUREMENTS: Outcomes were the number of days of absence from work because of low back pain, functional status (Roland Disability Questionnaire), and severity of pain (11-point numerical scale). RESULTS: The median number of days of absence from work over 6 months of follow-up was 58 days in the graded activity group and 87 days in the usual care group. From randomization onward, graded activity was effective after 50 days of absence from work (hazard ratio, 1.9 [95% CI, 1.2 to 3.2]; P = 0.009). The graded activity group was more effective in improving functional status and pain than the usual care group. The effects, however, were small and not statistically significant. CONCLUSIONS: Graded activity was more effective than usual care in reducing the number of days of absence from work because of low back pain. PMID- 14734330 TI - High-dose melphalan and autologous stem-cell transplantation in patients with AL amyloidosis: an 8-year study. AB - BACKGROUND: AL amyloidosis is a fatal disease resulting from tissue deposition of amyloid fibrils derived from monoclonal immunoglobulin light chains. Treatment with oral chemotherapy is minimally effective. OBJECTIVE: To test survival and organ response in a large sample of patients treated with high-dose intravenous melphalan (100 to 200 mg/m2) and autologous blood stem-cell transplantation. DESIGN: 8-year longitudinal analysis of clinical effectiveness. SETTING: University-affiliated specialty referral clinic. PATIENTS: 701 consecutive new patients with AL amyloidosis. INTERVENTION: High-dose chemotherapy and autologous stem-cell transplantation for patients who met eligibility requirements based on organ involvement and clinical status. MEASUREMENTS: Survival analysis of all patients evaluated and a detailed analysis of treatment outcome in the subgroup that received high-dose melphalan and stem-cell transplantation. RESULTS: Among 701 patients with AL amyloidosis, 394 (56%) were eligible for high-dose melphalan and stem-cell transplantation; 82 did not proceed with treatment because of patient choice or disease progression. Median survival of the 312 patients who initiated treatment was 4.6 years. A complete hematologic response, defined as no evidence of an underlying plasma cell dyscrasia 1 year after treatment, was achieved in 40% of patients and was associated with prolonged survival. Statistically significant improvements occurred in end-organ disease and were greater in patients with a complete hematologic response. Mortality rate within 100 days of treatment with high-dose melphalan and stem-cell transplantation was 13%; patients with cardiomyopathy had the highest mortality rates. CONCLUSIONS: Treatment of selected patients with AL amyloidosis by using high-dose melphalan and stem-cell transplantation resulted in hematologic remission, improved 5-year survival, and reversal of amyloid-related disease in a substantial proportion. PMID- 14734331 TI - Elevated plasma homocysteine level is an independent predictor of coronary heart disease events in patients with type 2 diabetes mellitus. AB - BACKGROUND: High plasma homocysteine level has been associated with increased risk for coronary heart disease (CHD) events in nondiabetic individuals, especially in those with previously diagnosed CHD. In persons with type 2 diabetes mellitus, the association between homocysteine level and cardiovascular disease may be stronger than that in nondiabetic individuals, but no large prospective studies have examined the relationship between homocysteine level and CHD mortality in persons with type 2 diabetes. OBJECTIVE: To investigate whether moderately elevated plasma homocysteine levels are independently related to increased incidence of fatal and nonfatal CHD events in persons with type 2 diabetes. DESIGN: Prospective study. SETTING: Finnish sample of patients with type 2 diabetes. PATIENTS: 462 men and 368 women who were 45 to 64 years of age at baseline. MEASUREMENTS: Coronary heart disease mortality and incidence of nonfatal myocardial infarction during the 7-year follow-up. RESULTS: Participants with plasma homocysteine levels of 15 micromol/L or more at baseline had a higher risk for CHD death than those with plasma homocysteine levels less than 15 micromol/L (26.1% and 13.5%, respectively; P = 0.005). The risks for all CHD events were 36.2% and 22.6%, respectively (P = 0.011). In Cox regression analyses, elevated plasma homocysteine level was significantly associated with CHD mortality (P < 0.001) and all CHD events (P = 0.002) even after adjustment for confounding variables, including creatinine clearance. In participants without myocardial infarction at baseline, moderate hyperhomocysteinemia was also associated with CHD mortality and all CHD events in univariate (P < 0.001 and P = 0.006, respectively) and multivariate Cox regression analyses (P < 0.001 and P = 0.004, respectively). CONCLUSIONS: In this large cohort of patients with type 2 diabetes, plasma homocysteine level was a strong and independent risk factor for CHD events. PMID- 14734332 TI - Resolution of left atrial thrombus after 6 months of anticoagulation in candidates for percutaneous transvenous mitral commissurotomy. AB - BACKGROUND: Resolution of left atrial thrombus after long-term oral anticoagulation enhances safe percutaneous transvenous mitral commissurotomy (PTMC); however, the short-term benefit has not been defined. OBJECTIVES: To estimate the resolution rate of left atrial thrombus among PTMC candidates after 6 months of oral anticoagulation and to determine its main predictors. DESIGN: Prospective cohort. SETTING: Community-based university medical center. PATIENTS: 219 PTMC candidates with thrombus demonstrated by multiplane transesophageal echocardiographic studies. MEASUREMENTS: The primary outcome was the status of the thrombus at the first 6-month follow-up; secondary measures were bleeding or thromboembolic complications. RESULTS: Among 219 PTMC candidates with left atrial thrombus (mean age [+/-SD], 39.6 +/- 7.4 years [range, 19 to 62 years]), complete resolution of thrombus, with an overall disappearance rate of 24.2% (95% CI, 18.5% to 29.9%), was demonstrated in 53 patients who subsequently underwent successful PTMC. In another 166 patients, the thrombus size was reduced by 24% (P < 0.001). No thrombus resolution was observed in the 27 patients with a left atrial body thrombus. Eighteen patients had minor bleeding. The significant predictors of thrombus resolution were a New York Heart Association class of 2 or less, a left atrial appendage thrombus size of 1.6 cm2 or less, a left atrial spontaneous echocardiographic contrast grade of 1 or less, and an international normalized ratio (INR) of at least 2.5. Patients with all of these predictors had a 94.4% chance of complete thrombus resolution (CI, 84.4% to 98.1%). CONCLUSIONS: After 6 months of oral anticoagulation, the left atrial thrombus disappeared in about a quarter of PTMC candidates so they could safely undergo PTMC. Less clinical severity, lower grading of the left atrial spontaneous echocardiographic contrast, a smaller thrombus, and a higher INR level predict thrombus resolution. PMID- 14734333 TI - Update in nephrology. PMID- 14734334 TI - Management of adult patients with persistent idiopathic thrombocytopenic purpura following splenectomy: a systematic review. AB - BACKGROUND: Treatment of chronic refractory idiopathic thrombocytopenic purpura is a dilemma because many patients have minimal symptoms, response to treatment is uncertain, and treatments may have serious adverse effects. PURPOSE: To determine the effectiveness of treatments for adult patients with idiopathic thrombocytopenic purpura who have not responded to splenectomy. DATA SOURCES: English-language reports from 1966 through 2003 that were retrieved from MEDLINE and Reference Update and bibliographies of retrieved articles. STUDY SELECTION: Articles reporting 5 or more total patients were reviewed to select eligible patients. Patients were eligible for inclusion if they were more than 16 years of age, had idiopathic thrombocytopenic purpura for more than 3 months, had a previous splenectomy, and had a platelet count less than 50 x 10(9) cells/L. DATA EXTRACTION: Patients were assessed for platelet count response, bleeding complications, duration of follow-up, and death. Complete remission was defined as a normal platelet count with no treatment for more than 3 months and for the duration of follow-up. DATA SYNTHESIS: 90 articles with 656 patients treated with 22 therapies met selection criteria. Azathioprine, cyclophosphamide, and rituximab had the most reported complete responses, but they were reported in only 41 to 109 patients. Reported complete response rates ranged from 17% to 27%, but 36% to 42% of patients had no response with these 3 treatments. Most reports described only platelet count responses; bleeding outcomes were reported in only 63 patients (10%). Only 111 (17%) of the 656 eligible patients had pretreatment platelet counts of less than 10 x 10(9) cells/L. No treatment method was reported in more than 20 patients. CONCLUSIONS: Evidence for the effectiveness of any treatment for patients with idiopathic thrombocytopenic purpura and persistent severe thrombocytopenia after splenectomy is minimal. Potentially effective treatments must be evaluated by randomized, controlled trials to determine both benefit and safety. PMID- 14734335 TI - The accuracy of the Ottawa knee rule to rule out knee fractures: a systematic review. AB - BACKGROUND: The Ottawa knee rule is a clinical decision aid that helps rule out fractures and avoid unnecessary radiography. PURPOSE: To summarize evidence about the accuracy of the Ottawa knee rule. DATA SOURCES: Relevant English- and non English-language articles were identified from PreMEDLINE and MEDLINE (1966 2003), EMBASE (1980-2003), CINAHL (1982-2003), BIOSIS (1990-2003), the Cochrane Library (2002, Issue 3), the Science Citation Index database, reference lists of included studies, and experts. STUDY SELECTION: Articles were included if they reported enough information to determine the sensitivity and specificity of the Ottawa knee rule for detecting fractures confirmed either radiologically or in combination with follow-up. DATA EXTRACTION: Two reviewers independently extracted data on study samples, the ways that the Ottawa knee rule was used, and methodologic characteristics of studies. DATA SYNTHESIS: Of 11 identified studies, 6 involving 4249 adult patients were considered appropriate for pooled analysis. The pooled negative likelihood ratio was 0.05 (95% CI, 0.02 to 0.23), the pooled sensitivity was 98.5% (CI, 93.2% to 100%), and the pooled specificity was 48.6% (CI, 43.4% to 51.0%). CONCLUSION: A negative result on an Ottawa knee rule test accurately excluded knee fractures after acute knee injury. However, because the rule is calibrated toward 100% sensitivity and actual fracture prevalences are usually low, large-scale, multicentered studies are still needed to establish the cost-effectiveness of routinely implementing the rule. PMID- 14734336 TI - Screening for thyroid disease: recommendation statement. AB - This statement summarizes the current U.S. Preventive Services Task Force (USPSTF) recommendations on screening for thyroid disease and updates the 1996 recommendations on this topic. The complete USPSTF recommendation 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), the National Guideline Clearinghouse (http://www.guideline.gov), and in print through the Agency for Healthcare Research and Quality Publications Clearinghouse (telephone, 800-358-9295; e-mail, ahrqpubs.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 the Web sites already mentioned. The recommendation statement and article are also available in print through the Agency for Healthcare Research and Quality Publications Clearinghouse. PMID- 14734337 TI - Screening for subclinical thyroid dysfunction in nonpregnant adults: a summary of the evidence for the U.S. Preventive Services Task Force. AB - BACKGROUND: Subclinical thyroid dysfunction is a risk factor for developing symptomatic thyroid disease. Advocates of screening argue that early treatment can prevent serious morbidity in individuals who are found to have laboratory evidence of subclinical thyroid dysfunction. PURPOSE: This article focuses on whether it is useful to order a thyroid function test for patients who have no history of thyroid disease and have few or no signs or symptoms of thyroid dysfunction. DATA SOURCES: A MEDLINE search, supplemented by searches of EMBASE and the Cochrane Library, reference lists, and a local database of thyroid related articles. STUDY SELECTION: Controlled treatment studies that used thyroid stimulating hormone (TSH) levels as an inclusion criterion and reported quality of life, symptoms, or lipid level outcomes were selected. Observational studies of the prevalence, progression, and consequences of subclinical thyroid dysfunction were also reviewed. DATA EXTRACTION: The quality of each trial was assessed by using preset criteria, and information about setting, patients, interventions, and outcomes was abstracted. DATA SYNTHESIS: The prevalence of unsuspected thyroid disease is lowest in men and highest in older women. Evidence regarding the efficacy of treatment in patients found by screening to have subclinical thyroid dysfunction is inconclusive. No trials of treatment of subclinical hyperthyroidism have been done. Several small, randomized trials of treatment of subclinical hypothyroidism have been done, but the results are inconclusive except in patients who have a history of treatment of Graves disease, a subgroup that is not a target of screening in the general population. Data on the adverse effects of broader use of L-thyroxine are sparse. CONCLUSION: It is uncertain whether treatment will improve quality of life in otherwise healthy patients who have abnormal TSH levels and normal free thyroxine levels. PMID- 14734338 TI - Absent from work: nature versus nurture. PMID- 14734339 TI - A time to listen. PMID- 14734340 TI - Implications of regional differences in spending. PMID- 14734341 TI - Implications of regional differences in spending. PMID- 14734342 TI - Implications of regional differences in spending. PMID- 14734343 TI - Implications of regional differences in spending. PMID- 14734344 TI - Implications of regional differences in spending. PMID- 14734345 TI - Implications of regional differences in spending. PMID- 14734346 TI - Implications of regional differences in spending. PMID- 14734347 TI - Quadriceps strength and osteoarthritis progression in malaligned and lax knees. PMID- 14734348 TI - Quadriceps strength and osteoarthritis progression in malaligned and lax knees. PMID- 14734349 TI - Quadriceps strength and osteoarthritis progression in malaligned and lax knees. PMID- 14734350 TI - Treatment of chronic hepatitis C in a state correctional facility. PMID- 14734351 TI - Is a strong quadriceps muscle bad for a patient with knee osteoarthritis? PMID- 14734352 TI - New treatments for growing scourge of brittle bones. PMID- 14734353 TI - Summaries for patients. A graded activity program for workers with disabling low back pain. PMID- 14734354 TI - Summaries for patients. Improved treatment of AL amyloidosis. PMID- 14734355 TI - Summaries for patients. High homocysteine levels increase risk for heart attacks in people with type 2 diabetes mellitus. PMID- 14734356 TI - Summaries for patients. Dissolving left atrial clots in patients with mitral stenosis. PMID- 14734357 TI - Summaries for patients. Treatment options for idiopathic thrombocytopenic purpura when splenectomy is ineffective. PMID- 14734358 TI - Summaries for patients. Screening for thyroid disease: a recommendation from the U.S. Preventive Services Task Force. PMID- 14734359 TI - Evidence for use of coronary stents. PMID- 14734360 TI - Ischemia-reperfusion induces G-CSF gene expression by renal medullary thick ascending limb cells in vivo and in vitro. AB - Ischemic acute renal failure involves not only the kidney but also extrarenal organs such as the bone marrow that produces inflammatory cells. By ELISA and RNase protection assays, we now show that renal ischemia-reperfusion increases serum concentrations of granulocyte macrophage colony-stimulating factor (G-CSF) protein and increases both G-CSF mRNA and protein in the ischemic kidney. In situ hybridization localized the increased G-CSF mRNA to tubule cells, including medullary thick ascending limb cells (mTAL), in the outer medulla. We also show that mTAL produce G-CSF protein and increase G-CSF mRNA after stimulation by reactive oxygen species in vitro. The production of G-CSF by the kidney after ischemia-reperfusion provides a means of communication from the injured kidney to the bone marrow. This supports the known inflammatory response to ischemia. PMID- 14734362 TI - Planning for chemical incidents by implementing a Delphi based consensus study. PMID- 14734363 TI - "Ischemia modified albumin": a new biochemical marker of myocardial ischaemia. PMID- 14734364 TI - "Reforming emergency care" and ambulance services. PMID- 14734365 TI - Emergency department response to the deliberate release of biological agents. AB - Bioterrorism is the use of biological agents outside the arena of war. Its purpose is to disrupt civilian life. This article investigates the role of the emergency department in the event of an act of bioterrorism. PMID- 14734366 TI - Domestic violence in emergency medicine patients. AB - Domestic violence is an important public health issue. This review seeks to inform doctors who care for patients who have suffered domestic violence. PMID- 14734367 TI - Advanced airway management in the emergency department: what are the training and skills maintenance needs for UK emergency physicians? AB - This article reviews the evidence for the training of emergency physicians in advanced airway management. PMID- 14734368 TI - Planning for chemical incidents by implementing a Delphi based consensus study. AB - This paper provides a practical approach to the difficulties surrounding planning for chemical incidents, based upon the results of a Delphi based consensus study. It is intended to offer advice, which can be implemented at regional and local prehospital and hospital level. The phases of the response that are covered include preparation, management of the incident, delivery of medical support during the incident, and recovery and support after the incident. PMID- 14734369 TI - Delphi based consensus study into planning for chemical incidents. AB - OBJECTIVE: To achieve consensus in all phases of chemical incident planning and response. DESIGN: A three round Delphi study was conducted using a panel of 39 experts from specialties involved in the management of chemical incidents. Areas that did not reach consensus in the Delphi study were presented as synopsis statements for discussion in four syndicate groups at a conference hosted by the Department of Health Emergency Planning Co-ordination Unit. RESULTS: A total of 183 of 322 statements had reached consensus upon completion of the Delphi study. This represented 56.8% of the total number of statements. Of these, 148 reached consensus at >94% and 35 reached consensus at >89%. The results of the process are presented as a series of synopsis consensus statements that cover all phases of chemical incident planning and response. CONCLUSIONS: The use of a Delphi study and subsequent syndicate group discussions achieved consensus in aspects of all phases of chemical incident planning and response that can be translated into practical guidance for use at regional prehospital and hospital level. Additionally, areas of non-consensus have been identified where further work is required. PMID- 14734370 TI - Role of "Ischemia modified albumin", a new biochemical marker of myocardial ischaemia, in the early diagnosis of acute coronary syndromes. AB - BACKGROUND: Diagnosis of cardiac ischaemia in patients attending emergency departments (ED) with symptoms of acute coronary syndromes is often difficult. Cardiac troponin (cTn) is sensitive and specific for the detection of myocardial damage but may not rise during reversible myocardial ischaemia. Ischemia Modified Albumin (IMA) has recently been shown to be a sensitive and early biochemical marker of ischaemia. METHODS AND RESULTS: This study evaluated IMA in conjunction with ECG and cTn in 208 patients presenting to the ED within three hours of acute chest pain. At presentation, a 12-lead ECG was recorded and blood taken for IMA and cardiac troponin T (cTnT). Patients underwent standardised triage, diagnostic procedures, and treatment. Results of IMA, ECG, and cTnT, alone and in combination, were correlated with final diagnoses of non-ischaemic chest pain, unstable angina, ST segment elevation, and non-ST segment elevation myocardial infarction. In the whole patient group, sensitivity of IMA at presentation for an ischaemic origin of chest pain was 82%, compared with 45% of ECG and 20% of cTnT. IMA used together with cTnT or ECG, had a sensitivity of 90% and 92%, respectively. All three tests combined identified 95% of patients whose chest pain was attributable to ischaemic heart disease. In patients with unstable angina, sensitivity of IMA used alone was equivalent to that of IMA and ECG combined. CONCLUSIONS: IMA is highly sensitive for the diagnosis of myocardial ischaemia in patients presenting with symptoms of acute chest pain. PMID- 14734371 TI - Medical management of deliberate drug overdose: a neglected area for suicide prevention? AB - OBJECTIVES: Overdoses account for a quarter of all suicides in England. The number of people who survive the immediate effects of their overdose long enough to reach medical attention, but who subsequently die in hospital is unknown. The aim of this study was to determine the proportion of overdose suicides dying in hospital and describe their sociodemographic characteristics. METHOD: Cross sectional analysis of routinely collected Hospital Episode Statistics data for England (1997 to 1999) to identify hospital admissions for overdose among people aged 12+ and the outcome of these admissions. RESULTS: Between 1997 and 1999 there were 233 756 hospital admissions for overdose, 1149 (0.5%) of these ended in the death of the patient such deaths accounted for 28% [corrected] of all overdose suicides and 8% [corrected] of total suicides. The median time between admission and death was three days (interquartile range one to nine days). The most commonly identified drugs taken in fatal overdose were paracetamol compounds, benzodiazepines, and tricyclic/tetracyclic antidepressants. CONCLUSION: Around a quarter of all overdose suicide deaths occur subsequent to hospital admission. Further more detailed research is required to discover if better pre-admission and in-hospital medical management of those taking serious overdoses may prevent some of these deaths. PMID- 14734372 TI - Proposed risk stratification in upper gastrointestinal haemorrhage: is hospitalisation essential? AB - AIMS: Patients with upper gastrointestinal haemorrhage (UGIH) are usually admitted to hospital regardless of the severity of the bleed. The aim of this study was to identify patients who could be safely managed without hospitalisation and immediate inpatient endoscopy. METHODS: Based on a literature review, a protocol was devised using clinical and laboratory data regarded as being of prognostic value. A retrospective observational study of consecutive patients who attended the emergency department (ED) with UGIH was conducted during one calendar month. RESULTS: Fifty four patients were identified of whom 44 (81%) were admitted. Twelve suffered an adverse event. One of the 10 patients (10%) initially discharged from the ED was later admitted. Strict implementation of the protocol would have resulted in safe discharge of a further 15 patients, (34% of those admitted), and a saving of an estimated 37 bed days per month. CONCLUSIONS: Patients at low risk from UGIH may be identified in the ED. If validated, this protocol may improve patient management and resource utilisation. PMID- 14734373 TI - Preconsultation use of analgesics on adults presenting to the emergency department with acute appendicitis. AB - OBJECTIVE: 279 cases of appendicitis were reviewed and compared for the difference between those patients who received pain medication before consulting a surgeon and those who were not treated with analgesics. METHODS: All patients aged 15 years and older who underwent appendicectomy for appendicitis between 1 July 2001 and 30 June 2002 were divided into group 1 (those who received preconsultaion use of analgesics) and group 2 (those who were not treated with analgesics). The following measures were compared: age, sex, symptom duration, initial vital signs, white blood cell counts, frequency of imaging studies, time to operative intervention, and operative findings. Continuous and categorical variables were analysed using t and chi(2) tests, respectively. RESULTS: A total of 279 patients were included for analysis. Patient details (age, sex, symptom duration) of the two study groups were similar. There was no statistically significant difference between group 1 and group 2 with respect to vital signs (systolic blood pressure, pulse rate, respiratory rate, body temperature), white blood cell counts, and frequency of imaging studies (ultrasound, computed tomography). There was no significant difference in the rate of perforated appendicitis between the two study groups although a shorter median time to operative intervention has been found in the group who received analegesia. CONCLUSION: The preconsultation use of analgesics in ED patients with a final diagnosis of appendicitis is not associated with a longer delay to operative intervention and is not associated with an increased rate of perforated appendicitis. PMID- 14734374 TI - Emergency ultrasound in the acute assessment of haemothorax. AB - AIMS: To evaluate thoracic ultrasound for the detection of haemothorax in patients with thoracic trauma against established investigations. METHODS: Thoracic ultrasound was performed as an extension of the standard focused assessment with sonography for trauma (FAST) protocol used at the Queen's Medical Centre for the assessment of adult patients with torso trauma. Fluid was sought in both pleural cavities using a hand portable ultrasound system by one of two non-radiologists trained in FAST. Findings were compared against subsequent investigations/procedures performed at the discretion of the attending emergency physician-supine chest radiography, intercostal drain, computed tomography, or thoracotomy. The sensitivity of the technique and the time taken to diagnosis for each investigation were recorded. RESULTS: Sixty one patients, 54 (89%) after blunt trauma, underwent thoracic ultrasound evaluation during the study. Twelve patients had a haemothorax detected by ultrasound and confirmed by computed tomography or by tube thoracostomy. Four haemothoraces detected on ultrasound were not apparent on trauma chest radiography. There were 12 true positives, 48 true negatives, no false positives, and one false negative scan. The sensitivity of ultrasound was 92% and specificity 100% with a positive predictive value of 100% and negative predictive value 98% for the detection of haemothorax after trauma. CONCLUSIONS: Emergency ultrasound of the chest performed as part of the primary survey of the traumatised patient can rapidly and accurately diagnose haemothorax and is a valuable tool to augment the immediate clinical assessment of these patients. PMID- 14734375 TI - A Delphi study to identify performance indicators for emergency medicine. AB - OBJECTIVES: The aim of this study was to identify performance indicators thought to reflect the quality of patient care in the emergency department. METHODS: A three round accelerated Expert Delphi study was conducted by email or fax. A panel of 33 experts drawn from the fields of emergency medicine, emergency nursing, professional service users, and patients were consulted. Participants were initially asked to propose performance indicators that reflected the quality of care given in the emergency department setting in the United Kingdom. In the second round these proposals were collated and scored using a 9 point Likert scale; those that had not reached consensus were returned for reconsideration in the light of group opinion. Those statements reaching a pre-defined consensus were identified. RESULTS: 224 performance indicators were proposed. Altogether 36 indicators reached consensus reflecting good departmental performance after round three; 24 of these were process measures. CONCLUSIONS: 36 potential indicators of good quality of care in the emergency department in the UK have been identified. PMID- 14734376 TI - Can additional experienced staff reduce emergency medical admissions? AB - BACKGROUND: Increases in emergency medical admissions are placing a strain upon hospitals throughout the world. The aim of the study was to evaluate the effect of a new post, the "A&E physician", upon emergency medical admissions to a hospital. METHODS: For six months the A&E physician workload was audited and a randomised controlled comparison undertaken. Days were randomised to "A&E physician present" or "A&E physician absent". The A&E physician recorded details of all patients referred for medical admission, any intervention made, and their disposal from A&E. Routine hospital data compared the mean daily number of medical admissions, non-medical admissions, and referrals to other hospitals. RESULTS: 124 days were randomised: 59 to A&E physician present, 65 to A&E physician absent. The A&E physician received 581 referrals and intervened in the management of 142 (24%). Of these, 80 were discharged home, apparently saving 1.4 admissions per day. However, randomised comparison showed that presence of the A&E physician was associated with a reduction of only 0.7 medical admissions per day (95% CI -1.7 to 3.2, p = 0.561), and an increase of 1.1 non-medical admissions (95% CI -0.2 to 2.3, p = 0.09) and 0.3 transfers to other hospitals per day (95% CI zero to 0.6, p = 0.09). Overall, hospital admissions were increased by 0.9 per day when the A&E physician was present (95% CI -1.8 to 3.6, p = 0.5). CONCLUSION: Despite receiving many referrals and discharging a substantial proportion of these patients home, the A&E physician did not significantly change emergency medical admissions and may have increased admissions to other specialties. PMID- 14734377 TI - Mild head injury: observation or computed tomography? Economic aspects by literature review and decision analysis. AB - OBJECTIVES: To compare the costs of two strategies for acute management of mild head injury: inhospital observation compared with acute computed tomography (CT) and home care. METHODS: Studies comparing costs for the two strategies that seem to have similar outcomes for patients were systematically reviewed. A decision tree analysis to compare the costs was also constructed, based on Swedish national costs and the risks found in a recent review on mild head injury complications. RESULTS: No studies were found that directly measured and compared risks, benefits, and costs of the two strategies. In the four studies retrieved, involving 4126 patients, the costs for hospital observation were factual, but a model was used to evaluate costs for the CT strategy. On average, costs were one third lower with CT. Also, the decision tree analysis demonstrated that the CT strategy was one third less expensive than inhospital observation. A sensitivity analysis showed this to be valid for nearly all cases. If these calculations hold true, a change of strategy could result in annual savings of pound 280,000/million inhabitants. CONCLUSION: The CT strategy seems to cost one third less than hospital observation. PMID- 14734378 TI - Wavelet analysis of pulse oximeter waveform permits identification of unwell children. AB - BACKGROUND: Children who are unwell often display signs of circulatory compromise. It has been observed that pronounced changes occur in the appearance of the photoplethysmogram (pulse oximeter tracing) in these children. The aim of the study was to discover if wavelet transforms can identify more subtle changes in the photoplethysmogram of children who are unwell. METHODS: Photoplethysmograms were obtained from children attending a paediatric accident and emergency department with clinical features suggestive of significant bacterial illness or circulatory compromise. Photoplethysmograms were also obtained from a control group of well children. Wavelet transforms were applied to the traces in an attempt to separate the two groups. RESULTS: 20 traces were obtained from unwell children and 12 from controls. Analysis of the entropy of the wavelet transform of the photoplethysmogram allows the differentiation of unwell children from controls (p = 0.00002). CONCLUSIONS: Wavelet transform of the photoplethysmogram offers the possibility of a rapid non-invasive method of screening children for significant illness. PMID- 14734379 TI - Facilities and equipment in district general hospitals in the Netherlands: are we prepared for the critically ill paediatric patients? AB - OBJECTIVE: To evaluate the inventory for initial treatment of critically ill children. DESIGN: Prospective study. SETTING: Paediatric emergency settings in 15 major district general hospitals. METHODS: Using an "expert opinion" created by paediatric intensivists, all hospitals were visited twice to check the inventory. Firstly, to examine the initial site of emergency care for children coming from outside the hospital. Secondly, to visit other emergency sites. A total score below 75% of the optimum was considered as not optimally equipped. MAIN RESULTS: Equipment to meet "respiratory problems" was considered by the experts as most essential. Seventy five per cent of all emergency sites scored below 75% (4 of 11 paediatric departments, 1 of 15 emergency rooms. The emergency room was in all aspects significantly better equipped than the paediatric department. Major differences and variations in the inventory were identified between all hospitals. CONCLUSIONS: Emergency rooms are better equipped to meet the needs of critically ill paediatric patients coming from outside the hospital than the paediatric departments. Paediatricians involved in the treatment of children who become critically ill during their stay in the hospital (the "indoor" patients), have less equipment and medication on the paediatric department at their disposal than on their emergency room. Obviously, emergency care on the paediatric wards should be equipped at the same level as in the emergency room because for both locations the "golden hour" is critically important in final outcome. PMID- 14734380 TI - A regional approach to improving night time supervision of accident and emergency departments. PMID- 14734381 TI - Emergency contraception. Has over the counter availability reduced attendances at emergency departments? AB - Legislation introduced in January 2001 has meant that progestogen only contraception is now available without prescription for women aged 16 years and over. Patient records of two emergency departments in the South East Thames region between 2000 and 2001 were reviewed and it was found that there was a 52% reduction in the number of women attending these emergency departments with requests for emergency contraception. These findings suggest that the legislation has meant that more women are getting their emergency contraception without prescription from pharmacies as compared with emergency departments. PMID- 14734382 TI - Simple and safe treatment of pretibial haematoma in elderly patients. AB - A simple and safe technique is described for evacuation of pretibial haematoma in elderly patients. A Yankauer sucker attached to the wall suction is used to evacuate the haematoma under local anaesthesia. This technique is used in the accident and emergency department and the ward. PMID- 14734383 TI - Managing change in the emergency department: a personal view. AB - This is a contribution to the occasional series on simulated interactive management PMID- 14734384 TI - Best evidence topic reports. Oxygen in acute uncomplicated myocardial infarction. AB - A short cut review was carried out to establish whether supplemental oxygen reduces mortality in patients with uncomplicated acute myocardial infarction. Altogether 290 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- 14734385 TI - Best evidence topic reports. Gastric lavage in paracetamol poisoning. AB - A short cut review was carried out to establish whether gastric lavage is better than activated charcoal in cases of poisoning with paracetamol. Altogether 63 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- 14734386 TI - Best evidence topic reports. Towards evidence based emergency medicine: best BETs from the Manchester Royal Infirmary. PMID- 14734387 TI - Best evidence topic reports. Oral corticosteroids in acute urticaria. AB - A short cut review was carried out to establish whether the addition of oral corticosteroids to antihistamines leads to a more rapid resolution of urticaria. Thirty nine papers were found using the reported search, of which two 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- 14734388 TI - Best evidence topic reports. Ice, pins, or sugar to reduce paraphimosis. AB - A short cut review was carried out to establish which of the ice glove technique, the multiple puncture technique, or the application of sugar was the best approach for paraphimosis reduction. Thirty three papers were found using the reported search, of which three 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- 14734389 TI - Best evidence topic reports. Is intravenous aminophylline better than intravenous salbutamol in the treatment of moderate to severe asthma? AB - A short cut review was carried out to establish whether intravenous salbutamol or intravenous aminophylline offers the quickest and least complicated treatment for patients with moderate to severe asthma not responding to inhaled therapy. Altogether 71 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- 14734390 TI - Best evidence topic reports. Hypertonic or isotonic saline in hypotensive patients with severe head injury. AB - A short cut review was carried out to establish whether hypertonic or isotonic saline improved the outcome most in patients with severe head injury. Altogether 66 papers were found using the reported search, of which three 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- 14734391 TI - JournalScan. PMID- 14734392 TI - Prehospital management and medical intervention after a chemical attack. AB - Chemical warfare agents are toxic weapons and emergency prehospital medical care providers should be well prepared, trained, and equipped to give response. Personnel need to be aware of the following medical issues regarding prehospital management of a chemical attack, event recognition, incident medical command and control, safety and protection, decontamination, isolation of the incident area (hot zone, warm zone, and cold zone), sampling and detection, psychological management, communication, triage, treatment, transportation, recovery activities and fatality management. During prehospital response, healthcare responders should provide self protection by wearing proper protective equipment and ensuring that the casualty is thoroughly decontaminated. Medical first responders are also responsible for performing triage in each zone of the incident area. Victims are triaged into four categories based on the need for medical care; immediate, delayed, minimal, and expectant. Finally, a medical emergency planning should be completed, and exercises conducted to test the system before an event occurs. PMID- 14734393 TI - The ABC of community emergency care: Introduction, series summary, the system of care. PMID- 14734394 TI - Prehospital cardiac arrest outcome is adversely associated with antiarrythmic agent use, but not associated with presenting complaint or medical history. AB - STUDY OBJECTIVE: This study associated survival from prehospital cardiac arrest to patient historical variables including presenting complaint, medications used, and medical history as a secondary end point in a trial evaluating the effect of bicarbonate administration. This raises issues concerning extensive prehospital historical assessment that may potentially delay care and transport. METHODS: This prospective multicentre trial enrolled 874 prehospital cardiac arrest patients encountered by urban, suburban, and rural emergency medical services. This group underwent conventional ACLS intervention followed by empiric early administration of sodium bicarbonate (1mEq/l). Survival was measured as the presence of vital signs on emergency department arrival. Data analysis used Student's t test, Fisher's exact test, chi2 with Pearson correlation, and logistic regression (p<0.05). Secondary end points were analysed including an association with common historical variables such as medical history, presenting complaint, or drugs used. RESULTS: The overall survival rate was 13.9% (110 of 793) of prehospital arrest patients. There was no correlation between historical factors, such as chief complaint or history of present illness (p = 0.277), medical history (p = 0.425), presence of specific disease conditions (p = 0.1125 0.956), or overall drug use (p = 0.002-0.9848). However, there was an adverse association between specific antiarrhythmic use (p = 0.003) and outcome. CONCLUSION: There is little relation of patient historical factors on the outcome from prehospital cardiac arrest raising issues of efficiency with history taking in prehospital care and transport. PMID- 14734395 TI - Ambulance emergency services for patients with coronary heart disease in Lancashire: achieving standards and improving performance. AB - OBJECTIVES: To examine the performance of a rural ambulance trust during two time periods, 1996/97 and 2001, with respect to achieving standards for ambulance journey times and delivery of clinical care for patients with suspected acute myocardial infarction (AMI). METHODS: Audit datasets on two cohorts of patients with chest pain and suspected AMI were assembled by the Lancashire Ambulance Service NHS Trust in north west England: 3706 patients during 1996/97 and 3423 in 2001. They were transported to four hospitals. The analyses covered journey timings, role of rapid response vehicles (RRV), and clinical procedures and the results were compared with prevailing national standards. RESULTS: Hourly and daily usage patterns were similar in the two periods. During 1996/97 the national rural target of 95% of response times being within 19 minutes was achieved (96% of calls), unlike the target of 50% within eight minutes (45.3% of calls). During 2001, 2684 (78.4%) calls had response times within eight minutes thus exceeding the revised national target of 75%. RRVs were despatched for 1214 (35.5%) of calls in 2001, and the mean response time (SD) for these vehicles was significantly shorter than for front line ambulances (0:05:53 (0:02:49) versus 0:07:04 (0:04:19), p<0.001), likewise the mean call to hospital time (0:32:38 (0:09:28) v 0:35:01 (0:12:09), p<0.001). Patients in 2001 were more likely to be given aspirin by the ambulance crews (74% of cases), while the rate of cannulation was lower. CONCLUSION: A significant improvement has been achieved in the performance of ambulance services in Lancashire since 1996, because of recently introduced strategies, notably RRVs, and in the presence of more demanding national standards and targets. PMID- 14734396 TI - Results of an evaluation of the effectiveness of triage and direct transportation to minor injuries units by ambulance crews. AB - OBJECTIVE: To evaluate triage and transportation to a minor injury unit (MIU) by emergency ambulance crews. METHODS: Ambulance crews in two services were asked to transport appropriate patients to MIU during randomly selected weeks of one year. During all other weeks they were to treat such patients according to normal practice. Patients were followed up through ambulance service, hospital and/or MIU records, and by postal questionnaire. Semi-structured interviews were undertaken with crews (n = 15). Cases transferred from MIU to accident and emergency (A&E) were reviewed. RESULTS: 41 intervention cluster patients attended MIU, 303 attended A&E, 65 were not conveyed. Thirty seven control cluster patients attended MIU, 327 attended A&E, 61 stayed at scene. Because of low study design compliance, outcomes of patients taken to MIU were compared with those taken to A&E, adjusted for case mix. MIU patients were 7.2 times as likely to rate their care as excellent (95% CI 1.99 to 25.8). Ambulance service job-cycle time and time in unit were shorter for MIU patients (-7.8, 95% CI -11.5 to -4.1); (-222.7, 95%CI -331.9 to -123.5). Crews cited patient and operational factors as inhibiting MIU use; and location, service, patient choice, job-cycle time, and handover as encouraging their use. Of seven patients transferred by ambulance from MIU to A&E, medical reviewers judged that three had not met the protocol for conveyance to MIU. No patients were judged to have suffered adverse consequences. CONCLUSIONS: MIUs were only used for a small proportion of eligible patients. When they were used, patients and the ambulance service benefited. PMID- 14734397 TI - Consensus on the prehospital approach to burns patient management. AB - Burns patients form a large group of trauma patients cared for by first aiders, ambulance staff, nurses, and doctors before reaching specialist care in hospital. Guidance for these important carers is often poor or confused and this engenders anxiety and detracts from optimal patient care. This paper outlines nine key steps in the initial management of burn patients in the prehospital environment based on current available evidence and a consensus of specialists from all disciplines caring for burns patients. The basis of care should be that simple things should always be performed well. PMID- 14734398 TI - Does the use of the Advanced Medical Priority Dispatch System affect cardiac arrest detection? AB - BACKGROUND: Cardiac arrest is the most widely recognised prehospital event that early intervention can directly affect. Chance of survival from this event decreases every minute that passes without treatment. To deliver a rapid ambulance response to these patients the early detection of cardiac arrest by control room staff is crucial. To achieve this, the London Ambulance Service (LAS) uses the Advanced Medical Priority Dispatch System. What impact has AMPDS had on identifying patients in cardiac arrest? Does compliance with AMPDS protocol influence the identification of patients in cardiac arrest? METHODS: A two stage study was undertaken. The first, compared cases coded as "cardiac arrest" and found by the responding ambulance to be in cardiac arrest before the implementation of AMPDS. This was compared with cases triaged as "cardiac arrest" and found to be in cardiac arrest across three years after AMPDS implementation. The second stage compared AMPDS compliance, over a 32 month period against the percentage of cardiac arrest calls that were found to be cardiac arrest upon the ambulance arrival. The correlation coefficient was calculated and analysed for statistical significance. FINDINGS: AMPDS resulted in a 200% rise in the number of patients accurately identified as suffering from cardiac arrest. A relation was identified between identification and AMPDS compliance (r(2) = 0.65, p = 0.001). DISCUSSION: The implementation of AMPDS increased accurate identification of patients in cardiac arrest. Additionally, the relation between factors identified suggests compliance with protocol is an important factor in the accurate recognition of patient conditions. PMID- 14734399 TI - A case of extreme hypercapnia: implications for the prehospital and accident and emergency department management of acutely dyspnoeic patients. PMID- 14734400 TI - Hypokalaemic thyrotoxic periodic paralysis in an Asian man in the United Kingdom. AB - A large number of ethnic Chinese and other oriental populations are living in the West because of the modern day migration of people. Hypokalaemic periodic paralysis attributable to thyrotoxicosis is a common presentation in an Asian emergency department. It is uncommon in the white communities. There is a difference in the genetic type in the different racial groups. Thyrotoxic features are often masked or absent. Life may be threatened because of severe hypokalaemia and therefore a prompt diagnosis of this condition in the certain ethnic group presenting with weakness and hypokalaemia is essential. Thyroid function studies are mandatory in these cases. PMID- 14734401 TI - Fatal descending necrotising mediastinitis. AB - Descending necrotising mediastinitis rarely develops and this variety of mediastinitis is a highly lethal disease. A case is reported of descending necrotising mediastinitis caused by an odontogenic infection. The importance is emphasised of prompt diagnosis and aggressive surgical mediastinal drainage for the survival of these patients. Most acute mediastinal infections result from oesophageal perforation, either secondary to oesophagoscopy or tumour erosion. Mediastinitis occasionally develops as descending necrotising mediastinitis originating from the complications of cervical or odontogenic infections. Descending necrotising mediastinitis usually has a fulminant course, leading commonly to sepsis and death. PMID- 14734402 TI - Troponin testing: beware pulmonary embolus. AB - Serum troponin estimation is widely used in the diagnosis and management of coronary syndromes, but it is possible to be misled by a positive result unless it is put carefully into clinical context. The serum troponin can be positive in pulmonary embolus and carries prognostic significance. A case report is presented and a review of the relevant literature. PMID- 14734403 TI - Emergency management of diabetic ketoacidosis. PMID- 14734404 TI - NICE head injury guidelines: cost implication for a district general hospital ("six scans to six figures"). PMID- 14734405 TI - Cost effective approach for emergency department investigation of deep vein thrombosis. PMID- 14734406 TI - The Kyoto commons--a tragedy? PMID- 14734409 TI - Common PGY-1: pro and con. PMID- 14734410 TI - It's no accident. PMID- 14734411 TI - Common PGY-1: pro and con. PMID- 14734413 TI - Eliminate trade barriers. PMID- 14734415 TI - Mercenary hypocrisy at CMAJ. PMID- 14734416 TI - Who was S. Weir Mitchell? PMID- 14734417 TI - More about hyperprolactinemia. PMID- 14734418 TI - More about hyperprolactinemia. PMID- 14734423 TI - Canadian researchers testing SARS vaccine in China. PMID- 14734428 TI - Stormy weather for Labour's NHS reforms. PMID- 14734429 TI - Global efforts to eradicate polio. PMID- 14734430 TI - Respiratory syncytial virus: pervasive yet evasive. PMID- 14734431 TI - Does the treatment of dyslipidemia affect the risk of depression or suicidal behaviour? PMID- 14734432 TI - A young man with an expanding neck mass. PMID- 14734433 TI - Multicentre, cluster-randomized clinical trial of algorithms for critical-care enteral and parenteral therapy (ACCEPT). AB - BACKGROUND: The provision of nutritional support for patients in intensive care units (ICUs) varies widely both within and between institutions. We tested the hypothesis that evidence-based algorithms to improve nutritional support in the ICU would improve patient outcomes. METHODS: A cluster-randomized controlled trial was performed in the ICUs of 11 community and 3 teaching hospitals between October 1997 and September 1998. Hospital ICUs were stratified by hospital type and randomized to the intervention or control arm. Patients at least 16 years of age with an expected ICU stay of at least 48 hours were enrolled in the study (n = 499). Evidence-based recommendations were introduced in the 7 intervention hospitals by means of in-service education sessions, reminders (local dietitian, posters) and academic detailing that stressed early institution of nutritional support, preferably enteral. RESULTS: Two hospitals crossed over and were excluded from the primary analysis. Compared with the patients in the control hospitals (n = 214), the patients in the intervention hospitals (n = 248) received significantly more days of enteral nutrition (6.7 v. 5.4 per 10 patient days; p = 0.042), had a significantly shorter mean stay in hospital (25 v. 35 days; p = 0.003) and showed a trend toward reduced mortality (27% v. 37%; p = 0.058). The mean stay in the ICU did not differ between the control and intervention groups (10.9 v. 11.8 days; p = 0.7). INTERPRETATION: Implementation of evidence-based recommendations improved the provision of nutritional support and was associated with improved clinical outcomes. PMID- 14734435 TI - Whither surgery in the treatment of gastroesophageal reflux disease (GERD)? PMID- 14734434 TI - Does universal comprehensive insurance encourage unnecessary use? Evidence from Manitoba says "no". AB - BACKGROUND: Many argue that "free" medical care leads to unnecessary use of health resources. Evidence suggests that user fees do discourage physician use, at least by those of low socioeconomic status. In this study, we compare health care utilization and health among socioeconomic groups to determine whether people of low socioeconomic status see physicians more than would be expected given their health status. METHODS: We examined the use of health care services (physicians and hospitals) by residents of Winnipeg, Manitoba, in 1999. The cost of physician services was drawn directly from the claims filed, and the cost of hospital services was estimated using the Case Mix Group and Day Procedure Group methods linked to resource intensity weights and Manitoba hospital costs. We used neighbourhood indicators of socioeconomic status from the 1996 census and measured health status by examining rates of premature mortality, acute myocardial infarction, hip fracture (1995-1999) and diabetes (1999). Using these measures, we compared health status and health care use of residents living in areas with low average household incomes with those living in areas with high average household incomes. All rates were age- and sex-adjusted across the groups. RESULTS: The province spent 44% more providing hospital and physician services to residents of Winnipeg neighbourhoods with the lowest household incomes (820 dollars/person annually v. 596 dollars/person for residents of the neighbourhoods with highest household incomes). However, expenditures were strongly related to health status. The 70% of the population on which the province spends 10% of its health care dollars scored well on all health indicators, and the 10% of the population on which 74% of the dollars are spent scored poorly. In each expenditure group, those with lower socioeconomic status had poorer health. In the highest expenditure group, those with lowest socioeconomic status had 82% higher premature mortality rates (23.0 v. 12.6 per 100,000 population) and 53% higher hip fracture rates (5.5 v. 3.6 per 100,000 population) than those with the highest socioeconomic status. Despite their poorer health, in each expenditure group, residents of the neighbourhoods with the lowest household incomes incurred physician expenditures that were similar to those of residents of wealthier neighbourhoods. INTERPRETATION: Most people use little health care; high-cost users are a small group of very sick people drawn from all neighbourhoods and all income groups. People living in areas with low average household incomes use fewer physician services than might be expected, despite their poor health status. PMID- 14734436 TI - A Canadian Agency for Public Health: could it work? PMID- 14734437 TI - Access to medicines and global health: will Canada lead or flounder? PMID- 14734443 TI - Imatinib therapy for patients with chronic myelogenous leukemia: are patients living longer? PMID- 14734438 TI - Adverse effects of antiretroviral therapy for HIV infection. AB - Long-term remission of HIV-1 disease can be readily achieved by combinations of antiretroviral agents. The suppression of plasma viral loads to less than the limit of quantification of the most sensitive commercially available assays (i.e., less than 50 copies/mL) and the coincident improvement in CD4 T cell counts is associated with resolution of established opportunistic infections and a decrease in the risk of new opportunistic infections. However, prolonged treatment with combination regimens can be difficult to sustain because of problems with adherence and toxic effects. All antiretroviral drugs can have both short-term and long-term adverse events. The risk of specific side effects varies from drug to drug, from drug class to drug class, and from patient to patient. A better understanding of the adverse effects of antiretroviral agents is of interest not only for HIV specialists as they try to optimize therapy, but also for other physicians who care for HIV-positive patients. PMID- 14734444 TI - A systematic review of molecular and biological tumor markers in neuroblastoma. AB - PURPOSE: The aim of this study was to conduct a systematic review, and where possible meta-analyses, of molecular and biological tumor markers described in neuroblastoma, and to establish an evidence-based perspective on their clinical value for the screening, diagnosis, prognosis, and monitoring of patients. EXPERIMENTAL DESIGN: A well-defined, reproducible search strategy was used to identify the relevant literature from 1966 to February 2000. RESULTS: A total of 428 papers studying the use of 195 different tumor markers in neuroblastoma were identified. Small sample sizes, poor statistical reporting, large heterogeneity across studies (e.g., in cutoff levels), and publication bias limited meta analysis to the area of prognosis only; MYCN, chromosome 1p, DNA index, vanillylmandelic acid:homovanillic acid ratio, CD44, Trk-A, neuron-specific enolase, lactate dehydrogenase, ferritin, and multidrug resistance were all identified as potentially important prognostic tools. CONCLUSIONS: This systematic review forms a knowledge base of the tumor markers studied thus far in neuroblastoma, and has identified some of the most important prognostic markers, which should be considered in future research and treatment strategies. Importantly, the review has also highlighted some general problems across primary tumor marker studies, in particular poor and heterogeneous reporting. These need to be addressed to allow better clinical interpretation and enable more appropriate evidence-based reviews in the future. In particular, collaboration of cancer research groups is needed to enable bigger sample sizes, standardize methods of analysis and reporting, and facilitate the pooling of individual patient data. PMID- 14734445 TI - Reducing the immune response to immunotoxin. PMID- 14734446 TI - Pretreatment with rituximab does not inhibit the human immune response against the immunogenic protein LMB-1. AB - PURPOSE: Rituximab, a humanized monoclonal antibody directed to the CD20 antigen present on B lymphocytes, could potentially abrogate the humoral immune response to murine monoclonal antibodies or immunotoxins by depleting antibody-producing B cells. EXPERIMENTAL DESIGN: A Phase II study of LMB-1, an immunotoxin targeting the Lewis Y tumor antigen, in combination with rituximab was conducted to test the hypothesis that rituximab could abolish or diminish the development of human antibodies to LMB-1. Five patients were treated in this study and received 375 mg/m(2) rituximab on days 1 and 7 followed by 45 micro g/kg/day LMB-1 on days 10, 12, and 14. The development of human antibodies against LMB-1 was detected using a serum neutralization and ELISA. RESULTS: All five of the patients had a total suppression of circulating CD20/CD19 B-cell population before the administration of the first dose of the immunotoxin. Before rituximab treatment, the mean percentage of CD20/CD19-positive B cells in the five treated patients was 19.8% (range, 4.5-29.8%) of the total peripheral lymphocytes. After two doses of rituximab, CD20/CD19-positive B lymphocytes constituted 50% regression of a lung nodule but progression of disease to the brain, whereas a second patient had a bona fide partial remission of a 3-cm diameter solitary lung nodule. CONCLUSIONS: LMI was nontoxic, improved immunological reactivity to melanoma cells, and showed evidence of clinical effectiveness (shrinkage of tumor) in 1 patient. Additional studies with LMI with added adjuvant materials, in melanoma and other cancers, appear warranted. PMID- 14734455 TI - Camptothecin analogs (irinotecan or topotecan) plus high-dose cyclophosphamide as preparative regimens for antibody-based immunotherapy in resistant neuroblastoma. AB - PURPOSE: We used high-dose cyclophosphamide plus topotecan/vincristine (CTV) or irinotecan (C/I) in patients with resistant neuroblastoma. The aim was to use a regimen with little risk to major organs to (a) achieve or consolidate remission in heavily treated patients and to (b) induce an immunological state conducive to passive immunotherapy with the murine 3F8 antibody. EXPERIMENTAL DESIGN: CTV and C/I included cyclophosphamide 140 mg/kg ( approximately 4200 mg/m(2)). With CTV, topotecan 2 mg/m(2) was infused i.v. (30 min) on days 1-4 (total, 8 mg/m(2)), and vincristine 0.067 mg/kg was injected on day 1. With C/I, irinotecan, 50 mg/m(2) was infused i.v. (1 h) on days 1-5 (total, 250 mg/m(2)). Mesna and granulocyte colony-stimulating factor were used. RESULTS: Twenty-nine patients received 38 courses of CTV, and 26 patients received 38 courses of C/I. All patients had previously received topotecan, a hemopoietic stem-cell transplant, and/or high dose cyclophosphamide. CTV and C/I caused myelosuppression of comparably prolonged duration as follows: absolute neutrophil counts <500/ micro l lasted 5 12 days in patients who had not previously received transplant and 7-21 days in patients who were post-transplant. Other significant toxicities included typhlitis (two CTV-treated patients, one C/I-treated patient) and hemorrhagic cystitis (one C/I-treated patient). Major responses were seen in 4 (15%) of 26 CTV and 4 (17%) of 24 C/I-treated patients with assessable disease. Bone marrow disease resolved in 5 (28%) of 18 CTV-treated patients and in 4 (27%) of 15 C/I treated patients. 3F8 after CTV or C/I was not blocked by neutralizing antibodies, consistent with the desired immunosuppressive effect of high-dose cyclophosphamide. CONCLUSIONS: CTV and C/I require transfusional and antibiotic support but otherwise entail tolerable morbidity. They have modest antineuroblastoma activity in heavily treated patients and are good preparative regimens for passive immunotherapy with monoclonal antibodies. PMID- 14734456 TI - Phase II study of SU5416, a small molecule vascular endothelial growth factor tyrosine kinase receptor inhibitor, in patients with refractory multiple myeloma. AB - PURPOSE: Increased bone marrow angiogenesis and vascular endothelial growth factor (VEGF) levels are of adverse prognostic significance in patients with multiple myeloma (MM). VEGF, a soluble circulating angiogenic molecule, acts via receptor tyrosine kinases, including VEGF receptor 2. SU5416 is a small molecule VEGF receptor 2 inhibitor. EXPERIMENTAL DESIGN: Adult patients with advanced MM were entered on a multicenter phase II study. RESULTS: Twenty-seven patients (median age 69, range 39-79), median 4 (0-10) lines of prior therapy, 14 with prior thalidomide therapy, received SU5416 at 145 mg/m(2) twice weekly i.v. for a median of two 4-week cycles (range 0.2-9). Grade 3/4 toxicities were rarely observed; the most frequent was thrombocytopenia (12%). Mild-to-moderate toxicities included nausea (63%), headache (56%), diarrhea, vomiting (both 37%), and fatigue (33%). There were three thromboembolic episodes and five cases of new onset hypertension. Two (7%) patients did not complete the first 4-week cycle of therapy because of adverse events (pneumonia and headache). There were no objective responses. Four patients had disease stabilization for >/==" BORDER="0">4 months. A decrease in median VEGF plasma levels was observed in patients with stable disease (n = 7) compared with patients with progressive disease (n = 5). Overall median survival was 42 weeks (range 3-92+). CONCLUSIONS: Although SU5416 had minimal clinical activity, signs of biological activity (decrease in plasma VEGF levels) suggest that angiogenic modulation may be of value in patients with MM. PMID- 14734457 TI - Cardiovascular safety profile of combretastatin a4 phosphate in a single-dose phase I study in patients with advanced cancer. AB - PURPOSE: The purpose of our study was to review and determine the cardiovascular safety profile of combretastatin A4 phosphate (CA4P) in a Phase I study in 25 patients with advanced solid tumors. EXPERIMENTAL DESIGN: CA4P was administered in a dose-escalating fashion starting at 18 mg/m(2) i.v. every 21 days, and the maximal dosage was 90 mg/m(2). Continuous evaluation included bedside blood pressure and pulse monitoring, 12-lead electrocardiogram (ECG) at fixed time points for measured QT interval determination, determination of the corrected QT interval (QTc) using Bazett's formula QTc = QT/(R-R interval)(1/2), and chart review. Pharmacodynamic correlations of CA4P dose, CA4P/CA4 area under the curve, and C(max) versus heart rate (HR), blood pressure, QT, and QTc intervals, over the first 4 h postdosing were analyzed. RESULTS: After CA4P administration, there were significant increases in QTc interval at the 3-h and 4-h time points [27.2 ms (P < 0.0001) and 30.8 ms (P < 0.0001), respectively] and HR at the 3- and 4-h time points [13.2 beats per minute (bpm; P < 0.01) and 15.1 bpm (P < 0.001), respectively]. Three of 25 patients had prolonged QTc intervals at baseline, whereas 15 (60%) of 25 and 18 (75%) of 24 patients had prolonged QTc intervals at 3 and 4 h. The slope of HR and QTc increasing as a function of time during the first 4 h was correlated to dose (in milligrams) of CA4P (P = 0.01 and r = 0.49 for HR, P = 0.005 and r = 0.55 for QTc) and to CA4 area under the curve (P = 0.04 and r = 0.41 for HR, P = 0.02 and r = 0.44 for QTc); blood pressure and uncorrected QTc interval dose-response correlations were not significant. Two patients had ECG changes consistent with an acute coronary syndrome within 24 h of CA4P infusion. CONCLUSIONS: CA4P prolongs the QTc interval. There was a temporal relationship with the CA4P infusion and with ECG changes consistent with an acute coronary syndrome in two patients. It is advisable that future trials with CA4P have eligibility guidelines limiting patients with known coronary artery disease or those with multiple coronary artery disease risk factors until more experience is gained regarding potential cardiovascular toxicity with this agent. PMID- 14734458 TI - Genomic organization, incidence, and localization of the SPAN-x family of cancer testis antigens in melanoma tumors and cell lines. AB - PURPOSE: Members of the SPAN-X (sperm protein associated with the nucleus mapped to the X chromosome) family of cancer-testis antigens are promising targets for tumor immunotherapy because they are normally expressed exclusively during spermiogenesis on the adluminal side of the blood-testis barrier, an immune privileged compartment. EXPERIMENTAL DESIGN AND RESULTS: This study analyzed the human SPANX genomic organization, as well as SPAN-X mRNA and protein expression in somatic and cancer cells. The SPANX family consists of five genes, one of which is duplicated, all located in a gene cluster at Xq27.1. From the centromere, the arrangement of the five SPANX genes mapped on one contiguous sequence is SPANXB, -C, -A1, -A2, and -D. Reverse transcription-PCR analyses demonstrated expression of SPAN-X mRNA in melanoma and ovarian cell lines, and virtual Northern analysis established SPANX gene expression in numerous cancer cell lines. Immunoblot analysis using polyclonal antisera raised against recombinant SPAN-X confirmed the translation of SPAN-X proteins in melanoma and ovarian tumor cell lines. The immunoreactive proteins migrated between M(r) 15,000 and M(r) 20,000 similar to those observed in spermatozoa. Immunoperoxidase labeling of melanoma cells and tissue sections demonstrated SPAN-X protein localization in the nucleus, cytoplasm, or both. Ultrastructurally, in melanoma cells with nuclear SPAN-X, the protein was associated with the nuclear envelope, a localization similar to that observed in human spermatids and spermatozoa. Significantly, the incidence of SPAN-X-positive immunostaining was greatest in the more aggressive skin tumors, particularly in distant, nonlymphatic metastatic melanomas. CONCLUSIONS: The data herein suggest that the SPAN-X protein may be a useful target in cancer immunotherapy. PMID- 14734459 TI - Hormone replacement therapy and lung cancer risk: a case-control analysis. AB - PURPOSE: To date, there are few published data regarding the use of hormone replacement therapy (HRT) and lung cancer risk. Therefore, we analyzed data regarding HRT use from a large case-control study designed to study genetic susceptibility to lung cancer to determine whether HRT affected risk of lung cancer. EXPERIMENTAL DESIGN: In a secondary analysis, we compared self-reported HRT use among 499 women with lung cancer and 519 healthy age-matched controls. RESULTS: HRT use was associated with an overall reduced risk of 34% [odds ratio (OR), 0.66; 95% confidence interval (CI), 0.51-0.89] of lung cancer, after adjusting for age, ethnicity, smoking status, education, body mass index, and menopausal status. The use of estrogen replacement therapy alone was associated with a 35% reduction in lung cancer risk (OR, 0.65; 95% CI, 0.47-0.89) and the use of combination therapy (estrogen and progestin) was associated with a 39% reduction in lung cancer risk (OR, 0.61; 95% CI, 0.40-0.92). HRT use was also associated with a statistically significantly reduced risk of lung cancer in current smokers (OR, 0.59; 95% CI, 0.38-0.92), but the risk estimates were not statistically significant in never (OR, 0.72; 95% CI, 0.37-1.40) or former smokers (OR, 0.73; 95% CI, 0.46-1.15). In addition, as the cigarette pack-years increased among ever smokers, the protective effect diminished, so that light smokers appeared to benefit the most from HRT use. Decreased lung cancer risks were also evident when the data were stratified by age, ethnicity, and body mass index. The joint effects of HRT use and mutagen sensitivity suggest that HRT use modifies lung cancer risk for genetically susceptible women. HRT use was also associated with a lower risk of death and improved survival compared with the women not taking HRT. To provide a possible biological mechanism to explain our findings, we compared plasma levels of insulin-like growth factor I in users and nonusers, and demonstrated that HRT use was associated with statistically significantly lower insulin-like growth factor I levels for both cases and controls compared with non-HRT users. CONCLUSIONS: These data suggest an association of HRT use with a decrease in lung cancer risk. However, there are several limitations to this secondary analysis, requiring that the data be viewed with caution, and confirmation is required in well-designed hypothesis driven studies. The biological role of HRT in lung cancer remains understudied, and only extensive research can yield new insights into the mechanisms underlying a protective effect of HRT for lung cancer. PMID- 14734460 TI - Associations between breast cancer susceptibility gene polymorphisms and clinicopathological features. AB - PURPOSE: Genetic polymorphisms may affect not only cancer development but also cancer progression, and as a result could influence cancer phenotypes. The aim of this study was to examine the relationship between breast cancer susceptibility gene polymorphisms and clinicopathological features. EXPERIMENTAL DESIGN: We genotyped 664 Korean primary breast cancer patients for 17 single-nucleotide polymorphisms (SNPs) in nine genes, using a high-throughput SNP scoring method. RESULTS: CYP1A1 codon 462 Ile/Val or Val/Val variants and the CYP1B1 codon 432 Leu/Val variant were found more in breast cancer patients 5 years and determined the status of the TP53 codon 72 polymorphism in DNA samples extracted from archived gastric tissues. RESULTS: The frequency of the arginine homozygous allele was positively correlated to patient age at baseline (P = 0.002). However, the age-related increase in the percentage of codon 72 arginine p53 was not correlated to the prognosis for gastric cancer patients. Multivariable analysis in patients who had surgery showed that baseline age may be inversely associated with patient survival (odds ratio, 1.1; 95% confidence interval, 1.0-1.2; P = 0.02). Furthermore, alcohol consumption may be associated with reduced survival (P = 0.06). CONCLUSIONS: These findings indicate that codon 72 arginine p53 may not be associated with a prolonged survival in patients with advanced gastric adenocarcinoma, but further study is needed to assess whether this polymorphism is associated with a late onset or slow progress of early gastric adenocarcinoma. PMID- 14734462 TI - Synchronous overexpression of epidermal growth factor receptor and HER2-neu protein is a predictor of poor outcome in patients with stage I non-small cell lung cancer. AB - PURPOSE: Despite maximal therapy, surgically treated patients with stage I non small cell lung cancer (NSCLC) are at risk for developing metastatic disease. Histopathologic findings cannot adequately predict disease progression, so there is a need to identify molecular factors that serve this purpose. Because the ErbB receptors play an important role in lung cancer progression, we analyzed the expression of epidermal growth factor receptor (EGFR), phosphorylated EGFR, transforming growth factor alpha (TGFalpha), and HER2-neu as potential prognostic factors in stage I NSCLC. EXPERIMENTAL DESIGN: Using immunohistochemical techniques, we retrospectively analyzed formalin-fixed, paraffin-embedded samples from 111 patients with resected pathological stage I NSCLC. Then we correlated these data with patient clinical outcome. RESULTS: Median follow-up was 69.3 months. EGFR overexpression (defined as >10% membranous staining) was found in 66 tumors (59.5%). It was significantly more common in T(2) tumors than in T(1) tumors (P = 0.001), and in more squamous cell carcinomas than in adenocarcinomas (P = 0.07). HER2-neu overexpression was found in 19 tumors (17.1%) and was significantly more common in adenocarcinomas than in squamous cell carcinomas (P = 0.035). Synchronous overexpression of EGFR and HER2-neu was found in 11 tumors (9.9%). Patients with these tumors had a significantly shorter time to recurrence (P = 0.006) and a trend toward shorter overall survival (P = 0.093). Phosphorylated EGFR and transforming growth factor alpha were detected but were not related to prognosis. CONCLUSIONS: Synchronous overexpression of EGFR and HER2-neu at the protein level predicts increased recurrence risk and may predict decreased survival in patients with stage I NSCLC. This suggests that important interactions take place among the different members of the ErbB family during tumor development and suggests a method for choosing targeted therapy. A prospective study is planned. PMID- 14734463 TI - Heterogeneous expression of interleukin-18 and its receptor in B-cell lymphoproliferative disorders deriving from naive, germinal center, and memory B lymphocytes. AB - PURPOSE: Dysregulated cytokine/cytokine receptor expression may occur in B-cell lymphoproliferative disorders. Little information is available on interleukin-18 receptor (IL-18R) and IL-18 expression in normal and malignant B cells. Our purpose was to investigate this issue in human naive, germinal center (GC) and memory B cells, and in their neoplastic counterparts. EXPERIMENTAL DESIGN: We have evaluated IL-18 expression and production in tonsil naive, GC, and memory B cells and in their presumed neoplastic counterparts by reverse transcription-PCR and ELISA. Moreover, IL-18Ralpha and beta expression was investigated in the same cells by reverse transcription-PCR, flow cytometry, and immunohistochemistry. RESULTS: We found that: (a) IL-18 mRNA was expressed in tonsil naive, GC, and memory B cells. Bioactive IL-18 was secreted by naive and GC, but not by memory B cells; (b) IL-18Ralpha and beta transcripts were expressed in the three B-cell subsets. IL-18Ralpha was detected on the surface of naive, GC, and memory B lymphocytes, and IL-18Rbeta was detected on GC and memory, but not naive, B cells; (c) mantle zone, follicular, marginal zone, Burkitt lymphoma (BL), and B cell chronic lymphocytic leukemia (B-CLL) cells expressed IL-18 mRNA. B-CLL and BL cells did not produce bioactive IL-18; and (d) lymphoma B cells displayed heterogeneous expression of either or both IL-18R chain mRNA. In contrast, B-CLL cells expressed both IL-18R chains at the mRNA and protein levels. CONCLUSIONS: Dysregulated expression of IL-18 and/or IL-18R in chronic B-cell lymphoproliferative disorders may sometimes contribute to tumor escape from the host immune system. PMID- 14734464 TI - Pharmacogenomic analysis of cytogenetic response in chronic myeloid leukemia patients treated with imatinib. AB - PURPOSE: To better understand the molecular basis of cytogenetic response in chronic myeloid leukemia patients treated with imatinib, we studied gene expression profiles from a total of 100 patients from a large, multinational Phase III clinical trial (International Randomized Study of IFN-alpha versus STI571). EXPERIMENTAL DESIGN: Gene expression data for >12,000 genes were generated from whole blood samples collected at baseline (before imatinib treatment) using Affymetrix oligonucleotide microarrays. Cytogenetic response was determined based on the percentage of Ph(+) cells from bone marrow following a median of 13 months of treatment. RESULTS: A genomic profile of response was developed using a subset of individuals that exhibited the greatest divergence in cytogenetic response; those with complete response (0% Ph(+) cells; n = 53) and those with minimal or no response (>65% Ph(+) cells; n = 13). A total of 55 genes was identified that were differentially expressed between these two groups. Using a "leave-one-out" strategy, we identified the optimum 31 genes from this list to use as our genomic profile of response. Using this genomic profile, we were able to distinguish between individuals that achieved major cytogenetic response (0 35% Ph(+) cells) and those that did not, with a sensitivity of 93.4% (71 of 76 patients), specificity of 58.3% (14 of 24 patients), positive predictive value of 87.7%, and negative predictive value of 73.7%. CONCLUSIONS: Interestingly, many of the genes identified appear to be strongly related to reported mechanisms of BCR-ABL transformation and warrant additional research as potential drug targets. The validity and clinical implications of these results should be explored in future studies. PMID- 14734465 TI - Predictive markers for late cervical metastasis in stage I and II invasive squamous cell carcinoma of the oral tongue. AB - PURPOSE: Patients with oral tongue carcinoma treated by intraoral excision only should be followed up carefully for cervical lymph node metastasis and salvaged immediately if found, because some patients have a more aggressive clinical course. The purpose of this study was to find useful markers for predicting late cervical metastasis in patients with stage I and II invasive squamous cell carcinoma of the oral tongue. EXPERIMENTAL DESIGN: We investigated clinicopathologic factors and immunohistochemical biomarkers predicting late cervical metastasis in surgical specimens from 56 patients with T(1-2)N(0)M(0) invasive squamous cell carcinoma of the oral tongue who did not undergo elective neck dissection. Histopathologic factors including tumor thickness, mode of invasion, Broders grade, total score of three different malignancy grading systems, eight other clinicopathologic parameters, and immunohistochemical expression of p53, cyclin D1, Ki-67, epidermal growth factor receptor, microvessel density, cyclooxygenase-2, MUC1, laminin-5 gamma2, E-cadherin, and beta-catenin were examined. All of the clinicopathologic factors and immunohistochemical expression of biomarkers were compared in terms of survival. RESULTS: In the univariate analysis, tumor thickness (P = 0.009), Broders grade (P = 0.017), nest shape (P = 0.005), mode of invasion (P < 0.001), Anneroth score (P = 0.029), Bryne score (P < 0.001), and E-cadherin expression (P = 0.003) were correlated with late cervical metastasis. Multivariate analysis on late cervical metastasis revealed that tumor thickness >4 mm, mode of invasion grade 3 or 4, and E-cadherin expression were independent factors. Late cervical metastasis was the only prognostic factor for overall survival (P = 0.002). CONCLUSIONS: Our results indicate that patients with stage I and II invasive squamous cell carcinoma of the oral tongue with tumor thickness >4 mm, mode of invasion grade 3 or 4, and low expression of E-cadherin should be considered a high-risk group for late cervical metastasis when a wait-and-see policy for the neck is adopted. PMID- 14734466 TI - Macrophage colony-stimulating factor-1 receptor expression is associated with poor outcome in breast cancer by large cohort tissue microarray analysis. AB - PURPOSE: Macrophage colony-stimulating factor-1 receptor (CSF-1R) is a transmembrane tyrosine kinase receptor, which is abnormally expressed in invasive breast cancer. Small cohort studies have demonstrated that increased expression of CSF-1R is associated with ipsilateral breast cancer recurrence. Correlation with survival has not been reported. Our aim was to further evaluate the role of CSF-1R in breast cancer, by studying the expression of CSF-1R in a large cohort of clinical specimens. EXPERIMENTAL DESIGN: Tissue microarrays containing 301 node-negative and 280 node-positive cases were used. Immunohistochemical staining was performed and correlated with overall survival, nodal status, and other clinicopathological data. RESULTS: CSF-1R expression was strongly associated with nodal status. Of the node-negative cases, 114 (38.9%) stained positive for CSF 1R, whereas 189 (67.5%) of the node-positive cases expressed CSF-1R (P < 0.0001). CSF-1R expression is also associated with larger tumor size (P = 0.02). Positive staining was strongly associated with decreased survival (P = 0.0003). Among node negative patients, CSF-1R expression was associated with decreased overall survival (P = 0.045), whereas among node-positive patients, it was not (P = 0.47). In multivariate analysis, CSF-1R was not independent of nodal status as a predictor of survival. CONCLUSIONS: CSF-1R expression is a strong predictor of poor outcome in nonmetastatic breast cancer. It is significantly more frequently expressed in patients with nodal involvement. Among the node-negative patients, it has a stronger association with survival than among the node-positive patients. Our findings support other preclinical findings that CSF-1R may be involved in local invasion and metastasis. Thus, this receptor may be an effective target for therapeutic agents. PMID- 14734467 TI - KIT (CD117)-positive breast cancers are infrequent and lack KIT gene mutations. AB - PURPOSE: KIT (CD117) is a transmembrane tyrosine kinase representing a target for STI571 (Glivec) therapy. Some KIT-overexpressing solid tumors have responded favorably to STI571, potentially because of the presence of KIT-activating mutations. EXPERIMENTAL DESIGN: To investigate the epidemiology of KIT overexpression and mutations, we investigated a series of 1654 breast cancers. All tumors were analyzed by immunohistochemistry in a tissue microarray format. RESULTS: KIT expression was always present in normal breast epithelium. However, cancer analysis revealed the only 43 of 1654 (2.6%) tumors were KIT-positive. KIT expression was more frequent in medullary cancer (9 of 47 positive; 19.1%) than in any other histological tumor subtype (P < 0.001). KIT expression was significantly associated with high tumor grade (P < 0.0001) but unrelated to pT and pN categories or patient survival. Mutation analysis of exons 2, 8, 9, 11, 13, and 17 was negative in 10 KIT-positive tumors. CONCLUSIONS: Overall, our data show that a high level of KIT expression occurs infrequently in breast cancer. KIT-positive breast cancers may not reflect "KIT up-regulation" because KIT is also expressed in normal breast epithelium. The lack of KIT mutations also argues against the therapeutic efficacy of STI571 in breast cancer. PMID- 14734468 TI - Correlation of osteopontin protein expression and pathological stage across a wide variety of tumor histologies. AB - PURPOSE: Osteopontin (OPN) is an integrin-binding protein overexpressed in various experimental models of malignancy and appears to be involved in tumorigenesis and metastasis. Although various studies have assessed OPN protein levels in several tumor types, a broad survey of OPN expression in human neoplasia under the same experimental conditions has not been carried out. EXPERIMENTAL DESIGN: We used immunohistochemistry to detect OPN in a selection of 350 human tumors and 113 normal tissues, from a variety of body sites, using stage-oriented human cancer tissue arrays. Tumors included malignancies from breast (26), ovary (22), endometrium (14), esophagus (10), stomach (11), pancreas (16), bile duct (1), liver (9), colon (20), kidney (53), bladder (33), prostate (28), head and neck (60), salivary glands (14), lung (17), skin (6), and brain (10). RESULTS: High cytoplasmic OPN staining was observed in 100% of gastric carcinomas, 85% of colorectal carcinomas, 82% of transitional cell carcinomas of the renal pelvis, 81% of pancreatic carcinomas, 72% of renal cell carcinomas, 71% of lung and endometrial carcinomas, 70% of esophageal carcinomas, 58% of squamous cell carcinomas of the head and neck, and 59% of ovarian carcinomas. Although OPN expression was identified in a good number of bladder, prostate, and brain tumors, the majority of 6 skin cancers, 11 of 14 salivary gland cancers, 2 thyroid carcinomas, and 23 of 26 breast cancers revealed low OPN positivity or were negative. When considering all sites, OPN expression significantly correlated with tumor stage (Spearman's correlation coefficient, P = 0.0002). OPN score and stage were also significantly correlated for specific cancer sites including bladder (P = 0.01), colon (P = 0.004), kidney (P = 0.0001), larynx (P = 0.035), mouth (P = 0.046), and salivary gland (P = 0.011). CONCLUSIONS: This study reports the broad distribution of OPN in human tumors from different body sites, suggesting involvement of this protein in tumor formation. The strong correlation between pathological stage and OPN across multiple tumor types suggests a role for OPN in tumor progression. PMID- 14734469 TI - BRAF mutation is frequently present in sporadic colorectal cancer with methylated hMLH1, but not in hereditary nonpolyposis colorectal cancer. AB - PURPOSE: The BRAF gene encodes a serine/threonine kinase and plays an important role in the mitogen-activated protein kinase signaling pathway. BRAF mutations in sporadic colorectal cancer with microsatellite instability (MSI) are more frequently detected than those in microsatellite stable cancer. In this study, we sought to compare the frequencies of BRAF mutations in sporadic colorectal cancer with MSI with those in hereditary nonpolyposis colorectal cancer (HNPCC). EXPERIMENTAL DESIGN: We analyzed BRAF mutations in 26 colorectal cancer cell lines, 80 sporadic colorectal cancers, and 20 tumors from HNPCC patients by DNA sequencing and sequence-specific PCR. The methylation status of the hMLH1 gene was measured by either sequencing or restriction enzyme digestion after NaHSO(3) treatment. RESULTS: We observed a strong correlation of BRAF mutation with hMLH1 promoter methylation. BRAF mutations were present in 13 of 15 (87%) of the colorectal cell lines and cancers with methylated hMLH1, whereas only 4 of 91 (4%) of the cell lines and cancers with unmethylated hMLH1 carried the mutations (P < 0.00001). Sixteen of 17 mutations were at residue 599 (V599E). A BRAF mutation was also identified at residue 463 (G463V) in one cell line. In addition, BRAF mutations were not found in any cancers or cell lines with K-ras mutations. In 20 MSI+ cancers from HNPCC patients, however, BRAF mutations were not detectable, including a subset of 9 tumors with negative hMLH1 immunostaining and methylated hMLH1. CONCLUSIONS: BRAF mutations are frequently present in sporadic colorectal cancer with methylated hMLH1, but not in HNPCC-related cancers. This discrepancy of BRAF mutations between sporadic MSI+ cancer and HNPCC might be used in a strategy for the detection of HNPCC families. PMID- 14734470 TI - Detection of micrometastatic disease and monitoring of perioperative tumor cell dissemination in primary operable breast cancer patients using real-time quantitative reverse transcription-PCR. AB - PURPOSE: We previously found a statistically significant number of cytokeratin 19 (CK19)+ cells in peripheral blood (PB) of stage IV breast cancer (BC) patients compared with those of healthy volunteers, using a quantitative real-time reverse transcription-PCR. We aimed to apply the technique on bone marrow (BM) of primary operable BC patients. Pre- and postoperative PB samples of these patients were further analyzed to investigate possible shedding of CK19+ cells during the operation. EXPERIMENTAL DESIGN: In 54 primary operable BC patients, we analyzed 50 BM samples taken preoperatively and 297 PB samples. PB samples were collected before surgery; immediately after surgery; on the first, second, and fifth day postoperatively; and one month postoperatively. RESULTS: In BM of controls and BC patients, we detected a median of 28 and 568 CK19+ cells/5 x 10(6) leukocytes, respectively (P < 0.001). In preoperative blood (B-1) samples, we measured a median of 109 CK19+ cells. Using the upper limit of 95% confidence interval of controls as cutoff, 74% and 52% of BM and (B-1), respectively were considered CK19+. There was no significant correlation between CK19+ cells in BM and (B-1) and classical prognostic factors. We found no significant difference between blood samples at different time points with respect to the average CK19+ cells. CONCLUSIONS: In primary BC patients, we detected high numbers of CK19+ cells in BM and PB (B-1) samples compared with controls. However, no significant correlation between the presence of CK19+ cells in BM and PB and classical prognostic factors was found. We detected no statistically significant influence of surgical manipulation on CK19+ cells. PMID- 14734471 TI - The hepatocyte growth factor regulatory factors in human breast cancer. AB - PURPOSE: Hepatocyte growth factor (HGF) stimulates tumor cell-cell interactions, matrix adhesion, migration, invasion, and angiogenesis. This factor is produced as an inactive precursor called pro-HGF, which requires proteolytic conversion, by HGF activator (HGFA) and matriptase, to evoke a biological response. Two new HGFA inhibitors, HAI-1 and HAI-2, inhibit the generation of biologically active HGF, through their interaction with HGFA. This study determined the expression of this HGF regulatory system in breast cancer. We examined HGF, the HGF receptor (c Met), HGFA, matriptase, and the activation inhibitors (HAI-1 and HAI-2), tissues from patients with breast cancer. EXPERIMENTAL DESIGN: Breast cancer tissue (n = 100) and normal background tissue (n = 20) was obtained immediately after surgery. The median follow-up for the patients was 72 months. HGF, c-Met, HGFA, matriptase-1, HAI-1, and HAI-2 expression was quantified using real-time quantitative PCR. The distribution of these factors in mammary tissues was also examined through immunohistochemistry. RESULTS: The breast cancer specimens expressed a significantly higher level of HGF, c-Met, HGFA, HAI-1, and HAI-2, but not matriptase, compared with the normal background tissues. Tumor tissues from node-positive patients expressed a higher level of HGFA than from the patients without nodal involvement. Interestingly, HAI-2 was expressed to a lower degree in positive nodes than that of the node-negative breast cancer tissues. HAI-1 and HAI-2 were both significantly reduced in grade 3 tumors compared with the well differentiated tumors. In addition, on comparison of Tumor-Node-Metastasis (TNM) classification groups, HAI-2 was also found to be statistically lower in the TNM 3 breast cancer group when compared with TNM groups 1 and 2, thus associated with a poor prognosis. CONCLUSIONS: This study shows that there are aberrant levels of HGF, c-Met, HGFA, HAI-1, and HAI-2 expressed in breast cancer tissues compared with background breast tissue. HAI-1 and HAI-2 are expressed to a significantly lower level in poorly differentiated breast tumors, and HAI-2 is also inversely correlated with nodal involvement and tumor spread. Overall a low level of HAI-2 in the breast cancer tissues was associated with an overall poor outlook. Therefore, the HGF regulatory system may have an important role in the progression of breast cancer. PMID- 14734472 TI - Identification of necrosis-associated genes in glioblastoma by cDNA microarray analysis. AB - PURPOSE: In the field of cancer research, there has been a paucity of interest in necrosis, whereas studies focusing on apoptosis abound. In neuro-oncology, this is particularly surprising because of the importance of necrosis as a hallmark of glioblastoma (GBM), the most malignant and most common primary brain tumor, and the fact that the degree of necrosis has been shown to be inversely related to patient survival. It is therefore of considerable interest and importance to identify genes and gene products related to necrosis formation. EXPERIMENTAL DESIGN: We used a nylon cDNA microarray to analyze mRNA expression of 588 universal cellular genes in 15 surgically resected human GBM samples with varying degrees of necrosis. Gene expression was correlated with the degree of necrosis using rank correlation coefficients. The expression of identified genes was compared with their expression in tissue samples from 5 anaplastic astrocytomas (AAs). Immunostaining was used to determine whether genes showing the most positive correlation with necrosis were increasingly expressed in tumor tissues, as grade of necrosis increased. RESULTS: The hybridization results indicated that 26 genes showed significant correlation with the amount of necrosis. All 26 genes had functions associated with either Ras, Akt, tumor necrosis factor alpha, nuclear factor kappaB, apoptosis, procoagulation, or hypoxia. Nine genes were positively correlated with necrosis grade, and 17 genes were negatively correlated with necrosis grade. There were significant differences in the median expression levels of 3 of the 26 genes between grade III necrosis GBM and anaplastic astrocytoma (AA) samples; all but 1 of the genes had elevated expression when comparing necrosis grade III with AA samples. Two factors, the ephrin type A receptor 1 and the prostaglandin E(2) receptor EP4 subtype, not previously considered in this context, were highlighted because of their particularly high (positive) correlation coefficients; immunostaining showed the products of these two genes to be localized in perinecrotic and necrotic regions and to be overexpressed in grade III GBMs, but not AAs. These two molecules also showed significant correlation with survival of GBM patients (P = 0.0034) in a combined model. CONCLUSIONS: The application of cDNA expression microarray analysis has identified specific genes and patterns of gene expression that may help elucidate the molecular basis of necrogenesis in GBM. Additional studies will be required to further investigate and confirm these findings. PMID- 14734473 TI - Altered N-myc downstream-regulated gene 1 protein expression in African-American compared with caucasian prostate cancer patients. AB - PURPOSE: The protein encoded by N-myc downstream-regulated gene 1 (NDRG1) is a recently discovered protein whose transcription is induced by androgens and hypoxia. We hypothesized that NDRG1 expression patterns might reveal a biological basis for the disparity of clinical outcome of prostate cancer patients with different ethnic backgrounds. EXPERIMENTAL DESIGN: Patients who underwent radical prostatectomy between 1990 and 2000 at Veterans Administration Medical Center of New York were examined. We studied 223 cases, including 157 African Americans and 66 Caucasians (T2, n = 144; >/=T3, n = 79; Gleason <7, n = 122; >/=7, n = 101). Three patterns of NDRG1 expression were identified in prostate cancer: (a) intense, predominately membranous staining similar to benign prostatic epithelium; (b) intense, nucleocytoplasmic localization; and (c) low or undetectable expression. We then examined the correlations between patients' clinicopathological parameters and different NDRG1 expression patterns. RESULTS: In this study of patients with equal access to care, African-American ethnic origin was an independent predictor of prostate-specific antigen recurrence (P < 0.05). We also observed a significant correlation between different patterns of NDRG1 expression and ethnic origin. Pattern 2 was less frequent in African Americans (21% versus 38%), whereas the reverse was observed for pattern 3 (60% in African Americans versus 44% in Caucasians; P = 0.03). This association remained significant after controlling for both grade and stage simultaneously (P = 0.02). CONCLUSIONS: Our data suggest that different NDRG1 expression patterns reflect differences in the response of prostatic epithelium to hypoxia and androgens in African-American compared with Caucasian patients. Further studies are needed to determine the contribution of NDRG1 to the disparity in clinical outcome observed between the two groups. PMID- 14734474 TI - Age-dependent prognostic effects of genetic alterations in glioblastoma. AB - PURPOSE: Although the genetic alterations in glioblastoma have been well characterized, reports regarding their prognostic effects have been inconsistent. EXPERIMENTAL DESIGN: In this series of 140 consecutive cases of glioblastoma treated at a single center, we analyzed the frequency, age dependency and prognostic effects of TP53 mutation, CDKN2A/p16 deletion, EGFR amplification, as well as loss of chromosome 1p, chromosome 10q, and chromosome 19q. The complete set of genetic alterations was available on 60 of 140 patients. RESULTS: In this cohort of glioblastoma cases, TP53 mutation was significantly associated with patient age. The prognostic effects of TP53 mutation, EGFR amplification, CDKN2A/p16 alterations, and loss of chromosome 1p were dependent on the age of the patient. CONCLUSIONS: This is the first observation that the prognostic effects of TP53, 1p, and CDKN2A/p16 alterations are dependent on patient age. These observations concerning the interactions of age and genetic changes in glioblastoma suggest that tumorigenic pathways to glioblastoma vary with the age of the patient and that future molecular marker studies should carefully evaluate the potential age-dependent prognostic effects of these biological variables. The inconsistent or negative prognostic effects of molecular markers reported in prior studies of glioblastoma may be because different effects at different ages may have resulted in a cancellation of an overall effect in the entire cohort. PMID- 14734475 TI - Impairment of both apoptotic and cytoprotective signalings in glioma cells resistant to the combined use of cisplatin and tumor necrosis factor alpha. AB - PURPOSE: Tumor necrosis factor (TNF)-alpha elicits two opposing effects, the induction of apoptosis and the transcription of antiapoptotic genes. We have recently shown that cisplatin sensitizes glioma cells to TNF-induced apoptosis, but only in some cell lines. To understand the mechanism involved in the different susceptibilities, we examined both the activation of caspases and cytoprotective signaling by TNF-alpha. EXPERIMENTAL DESIGN: Caspase activation was examined by estimating the cleavage of substrate peptides and by immunoblot to identify the cleavage of procaspases. Peptide inhibitors of caspases were used to reverse the cytotoxicity. The binding of TNF-alpha to the receptor was analyzed by flow cytometry. Nuclear factor (NF)-kappaB activation was assayed by the binding of NF-kappaB to oligonucleotides containing the consensus binding site. Interleukin (IL)-1beta, IL-6, IL-8, and manganous superoxide dismutase (MnSOD) were measured by enzyme-linked immunoassays. RESULTS: T98G and U87MG underwent apoptosis on treatment with cisplatin and TNF-alpha, but U373MG and A172 were resistant. Caspases 2, 3, and 6-10, but not caspases 1, 4, and 5, were activated in sensitive cells, and none were activated in resistant cells. The binding of TNF-alpha to the receptor was the same in all four of the cell lines. In the sensitive cells, NF-kappaB activation and the production of IL-1beta, IL 6, IL-8, and MnSOD were significantly elevated by TNF-alpha. However, in the resistant cells, the production of IL-1beta and IL-6 were specifically impaired in response to TNF-alpha. CONCLUSIONS: Our results indicate that both apoptotic and cytoprotective pathways are impaired in glioma cells that are resistant to treatment with cisplatin and TNF-alpha. PMID- 14734476 TI - In vivo suppression of hormone-refractory prostate cancer growth by inositol hexaphosphate: induction of insulin-like growth factor binding protein-3 and inhibition of vascular endothelial growth factor. AB - PURPOSE: Diet composition is an important etiologic factor in prostate cancer (PCA) growth and has significant impact on clinical PCA appearance. Because inositol hexaphosphate (IP6) is a dietary phytochemical present in cereals, soy, legumes, and fiber-rich foods, we evaluated efficacy of IP6 against PCA growth and associated molecular events. EXPERIMENTAL DESIGN: DU145 cells were injected into nude mice, and animals were fed normal drinking water or 1 or 2% IP6 in drinking water for 12 weeks. Body weight, diet, water consumption, and tumor sizes were monitored. Tumors were immunohistochemically analyzed for proliferating cell nuclear antigen, terminal deoxynucleotidyl transferase mediated nick end labeling, and CD31. Tumor-secreted insulin-like growth factor binding protein (IGFBP)-3 and vascular endothelial growth factor (VEGF) were quantified in plasma by ELISA. RESULTS: IP6 feeding resulted in suppression of hormone-refractory human prostate tumor growth without any adverse effect on body weight gain, diet, and water consumption during entire study. At the end of study, tumor growth inhibition by 1 and 2% IP6 feeding was 47 and 66% (P = 0.049 0.012) in terms of tumor volume/mouse and 40 and 66% (P = 0.08-0.003) in terms of tumor weight/mouse, respectively. Tumor xenografts from IP6-fed mice showed significantly (P < 0.001) decreased proliferating cell nuclear antigen-positive cells but increased apoptotic cells. Tumor-secreted IGFBP-3 levels were also increased up to 1.7-fold in IP6-fed groups. Additionally, IP6 strongly decreased tumor microvessel density and inhibited tumor-secreted VEGF levels. CONCLUSIONS: IP6 suppresses hormone-refractory PCA growth accompanied by inhibition of tumor cell proliferation and angiogenesis and increased apoptosis. IP6-caused increase in IGFBP-3 and decrease in VEGF might have a role in PCA growth control. PMID- 14734477 TI - Effective intravenous therapy of murine pulmonary metastases with an oncolytic herpes virus expressing interleukin 12. AB - PURPOSE: There currently is no therapy that enhances the survival of patients with distantly metastatic squamous cell carcinoma (SCC). Engineered herpes oncolytic viruses are effective therapeutic agents when delivered directly to tumors in animal models, but their efficacy in treating disseminated disease is poorly defined. EXPERIMENTAL DESIGN: We treated disseminated pulmonary SCC in mice with an interleukin (IL)-12-expressing oncolytic herpes virus (NV1042) or with the parent oncolytic virus (NV1023, IL-12 deficient) by i.v. tail vein administration. RESULTS: Lung IL-12 was 16.1 pg/mg and IFN-gamma was 4.3 pg/mg at day 1 after a single dose of NV1042 (5 x 10(7) plaque-forming units); levels of both were undetectable for NV1023. 5-Bromo-4-chloro-3-indolyl-beta-D galactopyranoside histochemistry demonstrated viral infection of disseminated pulmonary tumor nodules by both vectors at day 1, with sparing of adjacent alveolar cells. NV1042-treated lungs showed no surface nodules at day 12, in contrast to NV1023-treated (92 +/- 27 surface nodules) and PBS-treated (225 +/- 9 surface nodules) lungs. Significantly enhanced survival was observed in NV1042 treated animals compared with NV1023- and PBS-treated animals (log rank < 0.05). In animals with a low tumor burden, 100% of NV1042-treated, 70% of NV1023 treated, and none of the control animals achieved long-term survival. NV1042 efficacy was similar to NV1023 efficacy in animals depleted of CD4/CD8 T lymphocytes, showing that IL-12 expression enhances oncolytic activity through immune effects. Histology showed no cytopathic effects in non-tumor-bearing lung, brain, spleen, liver, and pancreas after completion of viral therapy. No animals demonstrated any visible side effects attributable to viral therapy. CONCLUSIONS: The i.v. delivery of an oncolytic herpes virus may achieve effective infection, oncolysis, and transgene expression at distant tumor sites. This approach to systemic therapy combining oncolysis with IL-12 immune stimulation led to significantly improved survival in animals with disseminated SCC. PMID- 14734478 TI - Chemosensitization of androgen-independent prostate cancer with neutral endopeptidase. AB - PURPOSE: We investigated whether neutral endopeptidase (NEP) could augment chemosensitivity to anticancer drugs by promoting protein kinase C (PKC)delta mediated mitochondrial apoptosis in prostate cancer (PC) cells. EXPERIMENTAL DESIGN: Human PC cell lines LNCaP and PC-3, and a normal prostate epithelial cell line (PrEC) were used. The protein expression was detected by Western blot analysis, and the protein turnover was determined by pulse-chase assay. Apoptotic ratio was measured by annexin V staining. RESULTS: Western blot analyses and pulse-chase assays showed that the specific NEP inhibitor CGS24592 decreased PKCdelta protein expression by promoting PKCdelta protein degradation in NEP expressing LNCaP cells. Conversely, recombinant NEP (rNEP) increased PKCdelta protein expression by delaying PKCdelta protein degradation in NEP-negative PC-3 cells. Apoptosis assays showed that rNEP promoted anticancer drug-induced apoptosis in PC-3 cells specifically through PKCdelta activity that mediated anticancer drug-induced mitochondrial change such as cytochrome-c release and caspase-9 activation. Of note, rNEP was able to increase PKCdelta protein expression predominantly in PC-3 cells rather than in PrEC cells. Treatment with rNEP before subtoxic concentrations of etoposide (0.1 micro M) significantly promoted mitochondrial apoptosis compared with only etoposide in PC-3 cells (P < 0.01) but not in PrEC cells. CONCLUSIONS: These results suggest that NEP enzyme activity contributes to anticancer drug-induced PC cell apoptosis dependent on PKCdelta-mediated mitochondrial events. More importantly, the combination of NEP with anticancer drugs may be a promising therapeutic modality because rNEP is able to augment chemosensitivity in androgen-independent PC with minimal toxicity in normal tissues. PMID- 14734479 TI - Celecoxib but not rofecoxib inhibits the growth of transformed cells in vitro. AB - PURPOSE: Nonsteroidal anti-inflammatory drugs reduce the risk of colorectal cancer. The cyclooxygenase (COX) pathway of arachidonic acid metabolism is an important target for nonsteroidal anti-inflammatory drugs. Increased expression of COX-2 was recently shown to be an important step in the multistep process of colorectal cancer carcinogenesis. The new COX-2-specific inhibitors offer the benefit of cancer protection without the gastrointestinal toxicity reported for the old drugs. The purpose of this study was to compare the growth effects of two specific COX-2 inhibitors, celecoxib (Pfizer, Inc., New York, NY), and rofecoxib (Merck, White House Station, NJ) in normal and transformed enterocytes. EXPERIMENTAL DESIGN: Cultures of normal rat intestinal epithelial cell line, IEC 18, vector control cells, c-K-ras, c-K-ras-bak, and antisense-bak derivatives were treated with different dosages of celecoxib (0-60 micro M) and rofecoxib (0 20 micro M). Cell cycle analysis and apoptosis were assessed by fluorescence activated cell sorting analysis. Protein expression was assessed by Western blot analysis and caspases 3 and 8 activities by ELISA. RESULTS: Celecoxib inhibited cell growth and induced apoptosis in a time- and dose-dependent manner. IEC18 parental cells were two to four times more resistant to celecoxib than ras, ras bak, and antisense bak transformed cells that overexpress the COX-2 protein. The induction of apoptosis by celecoxib involved the caspase pathways. Rofecoxib, up to its maximal concentration of 20 micro M, did not inhibit cell growth or induce apoptosis. CONCLUSIONS: Celecoxib may prove to be a very efficient component in the prevention and treatment of gastrointestinal tumors because it inhibits the growth of cancerous cells without affecting the growth of normal cells. PMID- 14734480 TI - Identification of genes with differential expression in acquired drug-resistant gastric cancer cells using high-density oligonucleotide microarrays. AB - PURPOSE: A major obstacle in chemotherapy is treatment failure due to anticancer drug resistance. The emergence of acquired resistance results from host factors and genetic or epigenetic changes in the cancer cells. The purpose of this study was to identify differentially expressed genes associated with acquisition of resistance in human gastric cancer cells. EXPERIMENTAL DESIGN: We performed global gene expression analysis in the acquired drug-resistant gastric cancer cell lines to the commonly used drugs 5-fluorouracil, doxorubicin, and cisplatin using Affymetrix HG-U133A microarray. The gene expression patterns of 10 chemoresistant gastric cancer cell lines were compared with those of four parent cell lines using fold-change and Wilcoxon's test for data analysis. RESULTS: We identified over 250 genes differentially expressed in 5-fluorouracil-, cisplatin , or doxorubicin-resistant gastric cancer cell lines. Our expression analysis also identified eight multidrug resistance candidate genes that were associated with resistance to two or more of the tested chemotherapeutic agents. Among these, midkine (MDK), a heparin-binding growth factor, was overexpressed in all drug-resistant cell lines, strongly suggesting that MDK might contribute to multidrug resistance in gastric cancer cells. CONCLUSIONS: Our investigation provides comprehensive gene information associated with acquired resistance to anticancer drugs in gastric cancer cells and a basis for additional functional studies. PMID- 14734481 TI - Telomerase-specific replication-selective virotherapy for human cancer. AB - PURPOSE: Replication-selective tumor-specific viruses present a novel approach for treating neoplastic disease. These vectors are designed to induce virus mediated lysis of tumor cells after selective viral propagation within the tumor. Telomerase activation is considered to be a critical step in carcinogenesis, and its activity is closely correlated with human telomerase reverse transcriptase (hTERT) expression. We investigated the antitumor effect of the hTERT-specific replication-competent adenovirus on human cancer cells. EXPERIMENTAL DESIGN: We constructed an adenovirus 5 vector [tumor- or telomerase-specific replication competent adenovirus (TRAD)], in which the hTERT promoter element drives expression of E1A and E1B genes linked with an internal ribosome entry site, and we examined the selective replication and antitumor effect in human cancer cells in vitro and in vivo. RESULTS: TRAD induced selective E1A and E1B expression in human cancer cells, but not in normal cells such as human fibroblasts. TRAD replicated efficiently and induced marked cell killing in a panel of human cancer cell lines, whereas replication as well as cytotoxicity was highly attenuated in normal human fibroblasts lacking telomerase activity. In nu/nu mice carrying s.c. human lung tumor xenografts, intratumoral injection of TRAD resulted in a significant inhibition of tumor growth. No evidence of TRAD was identified in tissues outside of the tumors, despite the presence of TRAD in the circulation. Moreover, TRAD replication in the distant, noninjected tumors was demonstrated. CONCLUSIONS: Our results suggest that the hTERT promoter confers competence for selective replication of TRAD in human cancer cells, an outcome that has important implications for the treatment of human cancers. PMID- 14734482 TI - Rapamycin inhibits the growth and metastatic progression of non-small cell lung cancer. AB - PURPOSE: Lung cancer has a dismal prognosis and comprises 5.5% of post-transplant malignancies. We explored whether rapamycin inhibits the growth and metastatic progression of non-small cell lung cancer (NSCLC). EXPERIMENTAL DESIGN: Murine KLN-205 NSCLC was used as the model tumor in syngeneic DBA/2 mice to explore the effect of rapamycin on tumor growth and metastastic progression. We also examined the effect of rapamycin on cell cycle progression, apoptosis, and proliferation using murine KLN-205 NSCLC cells and human A-549 NSCLC cells as targets. The in vivo and in vitro effects of cyclosporine and those of rapamycin plus cyclosporine were also investigated. RESULTS: Rapamycin but not cyclosporine inhibited tumor growth; s.c. tumor volume was 1290 +/- 173 mm(3) in untreated DBA/2 mice, 246 +/- 80 mm(3) in mice treated with rapamycin, and 1203 +/- 227 mm(3) in mice treated with cyclosporine (P < 0.001). Rapamycin but not cyclosporine prevented the formation of distant metastases; eight of eight untreated mice and four of six mice treated with cyclosporine developed pulmonary metastases whereas only one of six mice treated with rapamycin developed pulmonary metastases (P = 0.003). In vitro, rapamycin induced cell cycle arrest at the G(1) checkpoint and blocked proliferation of both KLN-205 and A-549 cells but did not induce apoptosis. Cyclosporine did not prevent cell cycle progression and had a minimal antiproliferative effect on KLN-205 and A-549 cells. CONCLUSIONS: The immunosuppressive macrolide rapamycin but not cyclosporine prevents the growth and metastatic progression of NSCLC. A rapamycin-based immunosuppressive regimen may be of value in recipients of allografts. PMID- 14734483 TI - Oral cavity and esophageal carcinogenesis modeled in carcinogen-treated mice. AB - PURPOSE: Squamous cell carcinoma of the oral cavity is one of the most common human neoplasms, and prevention of these carcinomas requires a better understanding of the carcinogenesis process and a model system in which cancer chemoprevention agents can be tested. We have developed a mouse model using the carcinogen 4-nitroquinoline 1-oxide (4-NQO) in the drinking water to induce tumorigenesis in the mouse oral cavity. EXPERIMENTAL DESIGN: 4-NQO was delivered by tongue painting or drinking water to two mouse strains, CBA and C57Bl/6. The incidences of oral cavity carcinogenesis were then compared. In addition, we examined the expression of some of the molecular markers associated with the process of human oral cavity and esophageal carcinogenesis, such as keratin (K) 1, K14, p16, and epidermal growth factor receptor, by immunohistochemistry. RESULTS: After treatment with 4-NQO in the drinking water, massive tumors were observed on the tongues of both CBA and C57Bl/6 female mice. Pathological analyses indicated that flat squamous dysplasias, exophytic papillary squamous tumors (papillomas), and invasive squamous cell carcinomas were present. Immunohistochemistry analyses showed that 4-NQO changed the expression patterns of the intermediate filament proteins K14 and K1. K14 was expressed in the epithelial suprabasal layers, in addition to the basal layer, in tongues from carcinogen-treated animals. In contrast, control animals expressed K14 only in the basal layer. Moreover, we observed more bromodeoxyuridine staining in the tongue epithelia of 4-NQO-treated mice. Reduced expression of the cell cycle inhibitor, p16, was observed, whereas 4-NQO treatment caused an increase in epidermal growth factor receptor expression in the mouse tongues. Interestingly, similar features of carcinogenesis, including multiple, large (up to 0.5 cm) exophytic papillary squamous tumors and invasive squamous cell carcinomas, increased bromodeoxyuridine staining, and increased K14 expression, were also observed in the esophagi of 4-NQO-treated mice. However, no tumors were observed in the remainder of digestive tract (including the forestomach, intestine, and colon) or in the lungs or livers of 4-NQO-treated mice. These results indicate that this murine 4-NQO-induced oral and esophageal carcinogenesis model simulates many aspects of human oral cavity and esophageal carcinogenesis. CONCLUSIONS: The availability of this mouse model should permit analysis of oral cavity and esophageal cancer development in various mutant and transgenic mouse strains. This model will also allow testing of cancer chemopreventive drugs in various transgenic mouse strains. PMID- 14734484 TI - Expression of the disintegrin metalloprotease, ADAM-10, in prostate cancer and its regulation by dihydrotestosterone, insulin-like growth factor I, and epidermal growth factor in the prostate cancer cell model LNCaP. AB - PURPOSE: The disintegrin metalloprotease ADAM-10 is a multidomain metalloprotease that is potentially significant in tumor progression due to its extracellular matrix-degrading properties. Previously, ADAM-10 mRNA was detected in prostate cancer (PCa) cell lines; however, the presence of ADAM-10 protein and its cellular localization, regulation, and role have yet to be described. We hypothesized that ADAM-10 mRNA and protein may be regulated by growth factors such as 5alpha-dihydrotestosterone, insulin-like growth factor I, and epidermal growth factor, known modulators of PCa cell growth and invasion. EXPERIMENTAL DESIGN: ADAM-10 expression was analyzed by in situ hybridization and immunohistochemistry in prostate tissues obtained from 23 patients with prostate disease. ADAM-10 regulation was assessed using quantitative reverse transcription PCR and Western blot analysis in the PCa cell line LNCaP. RESULTS: ADAM-10 expression was localized to the secretory cells of prostate glands, with additional basal cell expression in benign glands. ADAM-10 protein was predominantly membrane bound in benign glands but showed marked nuclear localization in cancer glands. By Western blot, the 100-kDa proform and the 60 kDa active form of ADAM-10 were synergistically up-regulated in LNCaP cells treated with insulin-like growth factor I plus 5alpha-dihydrotestosterone. Epidermal growth factor also up-regulated both ADAM-10 mRNA and protein. CONCLUSIONS: This study describes for the first time the expression, regulation, and cellular localization of ADAM-10 protein in PCa. The regulation and membrane localization of ADAM-10 support our hypothesis that ADAM-10 has a role in extracellular matrix maintenance and cell invasion, although the potential role of nuclear ADAM-10 is not yet known. PMID- 14734485 TI - Increased levels of tissue endostatin in human malignant gliomas. PMID- 14734487 TI - Proceedings of the Third International Conference on Recent Advances and Future Directions in Endocrine Manipulation of Breast Cancer: conference summary statement. PMID- 14734488 TI - Cross-talk between estrogen receptor and growth factor pathways as a molecular target for overcoming endocrine resistance. AB - Introduced more than 100 years ago, endocrine therapy is still the most important systemic therapy for all stages of estrogen receptor (ER) -positive breast tumors. A major clinical problem limiting the usefulness of this therapy is tumor resistance, either de novo or acquired during the course of the treatment. Relatively new discoveries emphasize the complexity of ER signaling and its multiple regulatory interactions with growth factor and other kinase signaling pathways. Both genomic (nuclear) and nongenomic (membrane and cytoplasmic) ER activities contribute to this intimate cross-talk, which is probably a fundamental factor in endocrine resistance. New targeted therapies, especially against the epidermal growth factor receptor/HER-2 pathway, should be carefully evaluated in more (bio)logical strategies to enable them to be exploited appropriately. A strategy of combining endocrine therapy (particularly tamoxifen) with these inhibitors, to circumvent de novo and acquired resistance, will be discussed. We will also emphasize open questions and future challenges in the dynamic research field of molecular ER biology from the endocrine therapy perspective. PMID- 14734489 TI - Adaptive hypersensitivity to estrogen: mechanism for sequential responses to hormonal therapy in breast cancer. AB - Clinical observations demonstrate that women with breast cancer often respond to subsequent endocrine manipulation after resistance to initial hormonal therapy develops. As a mechanistic explanation for these findings, we hypothesized that human breast tumors can adapt in response to the pressure exerted by endocrine therapy with development of hypersensitivity to estradiol. To understand the signaling pathways responsible, we examined estrogenic stimulation of cell proliferation in a model system and provided in vitro and in vivo evidence that long-term deprivation of estradiol (LTED) causes adaptive hypersensitivity. Even though the estrogen receptor alpha (ERalpha) is markedly up-regulated in LTED cells, the enhanced responses to estradiol do not appear to involve mechanisms acting at the level of transcription of estrogen-regulated genes. We found that ERalpha co-opts a classical growth factor pathway and induces rapid nongenomic effects that are enhanced in LTED cells. Estradiol binds to cell membrane associated ERs, physically associates with the adaptor protein Shc, and induces its phosphorylation. In turn, Shc binds Grb2 and Sos, which result in the rapid activation of mitogen-activated protein kinase. These nongenomic effects of estradiol produced biological effects, as evidenced by Elk-1 activation and by morphological changes in cell membranes. The mechanistic pathways involved in adaptive hypersensitivity suggest that inhibitors of the mitogen-activated protein kinase and phosphatidylinositol-3-OH kinase pathways might prevent the development of adaptive hypersensitivity and allow more prolonged efficacy of endocrine therapies. PMID- 14734490 TI - Nonendocrine pathways and endocrine resistance: observations with antiestrogens and signal transduction inhibitors in combination. AB - An increasing body of evidence demonstrates that growth factor networks are highly interactive with estrogen receptor signaling in the control of breast cancer growth. As such, tumor responses to antiestrogens are likely to be a composite of the estrogen receptor and growth factor-inhibitory activity of these agents, with alterations/aberrations in growth factor signaling providing a mechanism for the development of antiestrogen resistance. In this light, the current article focuses on illustrating the relationship between growth factor signaling and antiestrogen failure in our in-house tumor models of breast cancer and describing how we are now beginning to successfully target growth factor activity to improve the effects of antiestrogen drugs and to block aggressive disease progression. PMID- 14734491 TI - Data from the Arimidex, tamoxifen, alone or in combination (ATAC) trial: implications for use of aromatase inhibitors in 2003. AB - The third-generation aromatase inhibitors (AIs) have improved efficacy and safety versus tamoxifen for treatment of advanced breast cancer. Currently, anastrozole is the only third-generation AI with adjuvant therapy data in postmenopausal women. Initial and updated results from the Arimidex, Tamoxifen, Alone or in Combination (ATAC) trial (median follow-up, 47 months) confirm it to be more effective than tamoxifen for disease-free survival with several important tolerability benefits. As a result, there has been much debate about whether or not anastrozole should be used routinely to treat postmenopausal women with early breast cancer. In its review, the American Society of Clinical Oncology Health Services Research Committee agreed that the updated ATAC analyses provided a greater level of assurance, in terms of both toxicity and efficacy, for use of anastrozole in the adjuvant setting. However, pending 5-year data from ATAC and other trials of adjuvant AI use, adjuvant anastrozole was recommended by American Society of Clinical Oncology Health Research Committee for use only under certain circumstances, with 5 years of tamoxifen remaining the standard. Anastrozole should be the preferred AI in this setting; data from the ATAC trial should not be extrapolated to other members of the class. Despite this conservative recommendation, the overall risk:benefit profile from the ATAC trial favors anastrozole, and it is expected that a more favorable efficacy and adverse effect profile will be maintained. Anastrozole should, therefore, now be considered a valid alternative option to tamoxifen for adjuvant hormonal treatment in all postmenopausal women with hormone receptor-positive early breast cancer. PMID- 14734492 TI - Sequencing of endocrine therapy in postmenopausal women with advanced breast cancer. AB - The introduction of the nonsteroidal aromatase inhibitor (NS-AI) anastrozole as an alternative to tamoxifen for adjuvant therapy of women with resected hormone receptor-positive breast cancer has added a management category into which patients presenting with metastatic disease can be placed. There are now essentially three such categories: (a) tamoxifen sensitive (no prior AI); (b) tamoxifen resistant (no prior AI); and (c) NS-AI resistant (no prior tamoxifen). Well-conducted Phase III trials provide evidence for choosing first-line therapy for advanced disease in categories a and b. In tamoxifen-sensitive patients, one can choose either NS-AI, anastrozole, or letrozole. In tamoxifen-resistant patients, one can choose either of the NS-AIs, the steroidal AI exemestane, or the estrogen receptor down-regulator fulvestrant. The situation is quite different for patients in category c. There are no Phase III trials of agents in patients who have experienced disease progression on a NS-AI. Phase II data are available for exemestane and high-dose estrogens, and retrospective data are available for tamoxifen and fulvestrant. Additional clinical trials are needed to determine an optimal sequencing strategy. PMID- 14734493 TI - Cdk inhibitor p27Kip1 and hormone dependence in breast cancer. AB - p27Kip1 is an important regulator of the G1 to S transition. While a potent inhibitor of cyclin-dependent-kinase (Cdk)2, p27 is also involved in assembly of cyclin D/Cdk4 complexes. Although rarely mutated, p27 is functionally downregulated in many human cancers by mechanisms involving enhanced degradation, cytoplasmic mislocalization, and/or sequestration by cyclin D/Cdk complexes in response to oncogenic signals. Therefore, low levels and/or cytoplasmic localized p27 have been associated with enhanced malignancy and poor patient prognosis in many neoplasias including breast cancer. Recent data discussed below suggest that a threshold of p27 is required for response to antiestrogens and, conversely, that low levels predict for antiestrogen resistance. These results imply that hormone receptor-positive tumors with low and/or cytosolic p27 respond poorly to antiestrogens and should be considered for alternative therapeutic strategies. PMID- 14734494 TI - Prevention strategies with aromatase inhibitors. AB - Aromatase (estrogen synthetase) inhibitors are superior to tamoxifen in terms of both efficacy and toxicity in the treatment of advanced breast cancer and also in the neoadjuvant setting. Recent results from the Arimidex, Tamoxifen, Alone or in Combination adjuvant trial showed a marked reduction in contralateral primary breast cancer with anastrozole, an apparent prevention effect. A similar effect was seen in the MA.17 adjuvant trial comparing letrozole with placebo after 5 years of adjuvant tamoxifen. This has accelerated interest in aromatase inhibitors as primary preventive therapy. Two studies being conducted by the National Cancer Institute of Canada's Clinical Trials Group select women by virtue of mammographic breast density. The International Breast Cancer Intervention Study 2 trial randomizes women at elevated risk to anastrozole or placebo. Because of its steroidal structure, exemestane may be more effective than the nonsteroidal aromatase inhibitors and may protect bone and lipid metabolism from the effects of estrogen ablation. Elevated prostaglandin E2 levels from cyclooxygenase-2 induction by preinvasive and invasive breast lesions increase a number of tumor-promoting pathways, including aromatase, as well as angiogenetic, antiapoptotic, and others. Additive or synergistic effects between celecoxib, a cyclooxygenase-2 inhibitor, and exemestane have been demonstrated and have led to the National Cancer Institute of Canada's Clinical Trials Group MAP.3 trial, which will randomize women at elevated risk to placebo or to exemestane with or without celecoxib. The efficacy and long-term toxicity data from the aromatase inhibitor prevention trials, and the identification of risk profiles from trial results, are awaited with interest. PMID- 14734495 TI - Breast cancer: the protective effect of pregnancy. AB - It has been firmly established in epidemiological studies that early full-term pregnancy affords lifetime protection against the development of breast cancer. This phenomenon can be mimicked in rat and mouse models of mammary cancer in which the hormones estrogen and progesterone are given for 21 days. Carcinogen induced proliferation is blocked as a consequence of hormone pretreatment. Among several genes implicated by molecular studies to be differentially expressed is the tumor suppressor gene p53. Both immunohistochemical and Western blot studies indicate that p53 protein expression is increased in hormone-pretreated mice and rats. The p53-regulated gene p21Cip1 is also increased concomitantly with p53. To test directly the causative role of p53 in conferring a protective phenotype, we examined the hormone-induced protective effect in BALB/c p53 null mammary epithelium. In the mammary epithelium, the absence of p53 gene expression abrogated the protective effect of prior pregnancy. The tumor incidence curves were superimposable in p53 null mammary epithelium that were treated with 7,12 dimethylbenzanthracene or pregnancy plus 7,12-dimethylbenzanthracene. These results demonstrate that p53 plays a pivotal role in hormone-induced protection and raises the question of the mechanisms by which the steroid hormones, estrogen and progesterone, functionally activate p53. PMID- 14734496 TI - Genomic approaches to understanding mammary tumor progression in transgenic mice and responses to therapy. AB - Scores of genetically engineered mice have been generated in the quest to understand mechanisms of breast cancer development and progression. More recently, there has been a growing trend for using such models for testing various therapeutic strategies and agents. The application of these mouse models for these purposes requires that they be characterized in ways that demonstrate they possess important similarities to human breast cancer. In particular, detailed comparisons of the features of the models to human breast cancer must include attention to the histological phenotypes, chromosomal and molecular alterations, and the predictive value of the models for preclinical testing. Whereas these models have become important tools for the study of breast cancer, the great majority of existing mouse mammary cancer models develop tumors that are estrogen receptor negative, with relatively few models demonstrating metastatic spread to the lungs, and none developing metastases to bone. This review focuses on recent studies using genomic approaches to further understand the oncogenic processes occurring in mouse models of mammary cancer and to compare these changes with those identified in human breast cancer. Gene expression profiling is being applied to help define pharmacological responses that occur in vivo. Detailed genomic analyses will provide important information for selecting models for specific experimental purposes, contribute to the understanding of oncogene-specific expression signatures and potential therapeutic targets, and further define mechanisms of chemoprevention and chemotherapy. PMID- 14734497 TI - Neoadjuvant endocrine therapy as a drug development strategy. AB - The aromatase inhibitors offer both toxicity and efficacy advantages over tamoxifen, but to date, their overall impact on breast cancer outcomes has been modest. Advanced breast cancer remains incurable, and for early stage disease, an improvement in survival with AI versus tamoxifen has yet to be demonstrated. Resistance to endocrine manipulation is at the core of the problem and must be overcome to make additional progress. A number of signal transduction inhibitors (STIs) are now under development as endocrine resistance modulators, including those targeting cyclooxygenase-2, HER1 and/or 2 kinase, mTOR, and farnesyl transferase. Developing STIs for this indication is a challenge, however, because we still do not have a clear understanding of the molecular basis of resistance. A complete understanding could translate into a series of endocrine therapy/STI combinations that would be tailored according to the biology of the individual tumor to achieve optimal efficacy and safety. The development of this strategy will require the ability to diagnose resistance mechanisms on a tumor-by-tumor basis, and this can only be attained through careful clinical investigation. Neoadjuvant endocrine therapy is an appealing context to conduct research in this area because clinical outcomes can be obtained within a few months of treatment, and repeated tumor sampling for biomarker analysis (pharmacodynamic tumor profiling) can be readily achieved. However, the optimal clinical investigative approaches, analytical techniques, and appropriate surrogate end points have yet to be identified and are the subject of several ongoing or planned clinical studies. PMID- 14734498 TI - Regulatory nodes that integrate and coordinate signaling as potential targets for breast cancer therapy. AB - Blockade of the estrogen receptor (ER) with antiestrogens and aromatase inhibitors is effective in the treatment of breast cancer. Why ER plays such a dominant role in breast cancer and represents such an excellent target remains to be defined. The ability of ER to respond to multiple inputs and to control expression of multiple downstream genes may be one of the reasons why ER is such a powerful target for breast cancer treatment. The recent modest performance of a number of targeted therapies in breast cancer has raised the question whether we will ever develop therapies that have such success as antiestrogens. Targeted therapies tend to inhibit a single pathway that is probably altered in only a subset of patients. Even within this subset, only a limited number of patients respond. The evidence that virtually all pathways can cross-talk and that they exhibit several layers of redundancy reveals a complexity of signaling networks that may defy the generation of targeted therapies with efficacy similar to antiestrogens. However, there are clearly regulatory nodes that can integrate multiple upstream inputs and elicit diverse downstream outputs. We provide evidence and rationales for integrins, insulin receptor substrates (IRSs), and cyclin D1 as potential therapeutic targets. These proteins, similar to ER, can integrate and coordinate multiple signals in breast cancer cells and thus mediate diverse aspects of breast cancer progression. New treatment targets will emerge in light of more global models of signal transduction that fully integrate all aspects of cell biology such as the role of the extracellular matrix and will hopefully result in the development of targeted therapies that show efficacy similar to antiestrogens. PMID- 14734499 TI - Malignant mammary cells acquire independence from extracellular context for regulation of estrogen receptor alpha. AB - Interactions between luminal epithelial cells and their surrounding microenvironment govern the normal development and function of the mammary gland. Alterations of these interactions can induce abnormal intracellular signaling pathways that affect the development and progression of breast tumors. One critical component of mammary gland development, as well as breast cancer progression, is the expression of estrogen receptors. In a previous study using cultured nonmalignant mammary epithelial cells, we found that the basement membrane molecules, laminin-1 and collagen-IV, were involved in maintenance of estrogen receptor (ER) alpha expression, and that this response could be interfered with by disrupting cell-extracellular matrix adhesion. Here we use phenotypically normal mammary epithelial SCp2 cells to dissect the promoter region of the ERalpha that is involved in the selective response to basement membrane. We also analyze the alteration of this response in SCg6 cells, a malignant cell line that shares a common lineage with the SCp2 cells, to provide insight into the relative overexpression of ERalpha and the unresponsiveness to basement membrane regulation found in those malignant cells. Evidence is presented to show the relevance of the cross-talk between different signaling pathways in the constitution of a functional tissue organization and how this integration may be disrupted in the malignant phenotype. PMID- 14734500 TI - Bringing reality to drug-eluting stents. PMID- 14734501 TI - Endothelin expression and the progression of heart failure: exemplifying the vagaries of therapeutic development. PMID- 14734502 TI - Microvasculature in acute myocardial ischemia: part I: evolving concepts in pathophysiology, diagnosis, and treatment. PMID- 14734503 TI - Manipulating cardiac contractility in heart failure: data from mice and men. PMID- 14734504 TI - Endothelial dysfunction in pulmonary hypertension. PMID- 14734505 TI - Images in cardiovascular medicine. Intramyocardial hemorrhage caused by myocardial contusion. PMID- 14734506 TI - Arterial stiffness and pulse wave velocity: problems with terminology. PMID- 14734507 TI - Anatomic considerations in therapeutic arteriogenesis for cerebral ischemia. PMID- 14734508 TI - Electrical resychronization of failing right ventricle. PMID- 14734509 TI - Covered stent septal ablation for hypertrophic obstruction cardiomyopathy. PMID- 14734510 TI - Differential modulation of caveolin-1 expression in cells of the vasculature by statins. PMID- 14734511 TI - Nebulous imaging. PMID- 14734512 TI - Catheter-directed thrombolysis for the treatment of symptomatic deep vein thrombosis. PMID- 14734513 TI - Prognosis of atrial fibrillation in congestive heart failure. PMID- 14734515 TI - Report from the 100th Cardiovascular and Renal Drugs Advisory Committee meeting: US Food and Drug Administration: December 8-9, 2003 Gaithersburg, MD. PMID- 14734516 TI - Plasticity of human adipose lineage cells toward endothelial cells: physiological and therapeutic perspectives. AB - BACKGROUND: Adipose tissue development and remodeling are closely associated with the growth of vascular network. We hypothesized that adipose tissue may contain progenitor cells with angiogenic potential and that therapy based on adipose tissue-derived progenitor cells administration may constitute a promising cell therapy in patients with ischemic disease. METHODS AND RESULTS: In mice, cultured stromal-vascular fraction (SVF) cells from adipose tissue have a great proangiogenic potential, comparable to that of bone marrow mononuclear cells in the mouse ischemic hindlimb model. Similarly, cultured human SVF cells differentiate into endothelial cells, incorporate into vessels, and promote both postischemic neovascularization in nude mice and vessel-like structure formation in Matrigel plug. In vitro, these cells represent a homogeneous population of CD34- and CD13-positive cells, which can spontaneously express the endothelial cell markers CD31 and von Willebrand factor when cultured in semisolid medium. Interestingly, dedifferentiated mature human adipocytes have the potential to rapidly acquire the endothelial phenotype in vitro and to promote neovascularization in ischemic tissue and vessel-like structure formation in Matrigel plug, suggesting that cells of endothelial and adipocyte phenotypes may have a common precursor. CONCLUSIONS: This study demonstrates, for the first time, that adipocytes and endothelial cells have a common progenitor. Such adipose lineage cells participate in vascular-like structure formation in Matrigel plug and enhance the neovascularization reaction in ischemic tissue. These results also highlight the concept that adipose lineage cells represent a suitable new cell source for therapeutic angiogenesis in ischemic disease. PMID- 14734517 TI - Randomized, controlled dose-ranging study of the selective adenosine A2A receptor agonist binodenoson for pharmacological stress as an adjunct to myocardial perfusion imaging. AB - BACKGROUND: Dipyridamole and adenosine cause frequent side effects as a result of nonspecific adenosine receptor stimulation. Selective agonism of the adenosine A2A receptor should result in a similar degree of coronary vasodilation (and thus similar perfusion images) with fewer side effects. METHODS AND RESULTS: In a multicenter, randomized, single-blind, 2-arm crossover trial, 240 patients underwent 2 single photon emission computed tomographic (SPECT) imaging studies in random order, first after pharmacological stress with adenosine and a second study with the selective adenosine A2A receptor agonist binodenoson, using 1 of 4 dosing regimens. Safety, tolerability, and SPECT image concordance between the 2 agents were examined. Exact categorical agreement in the extent and severity of reversible perfusion defects ranged from 79% to 87%, with kappa values from 0.69 to 0.85, indicating very good to excellent agreement between binodenoson and adenosine. The risk of any safety event/side effect was significantly lower with any dose of binodenoson than with adenosine (P< or =0.01) because of a dose related reduction in subjective side effects, as objective events were infrequent. There was a reduction in the severity of chest pain, dyspnea, and flushing in all binodenoson doses compared with adenosine (P<0.01), and the magnitude of severity reduction was dose-related. CONCLUSIONS: The selective adenosine A2A receptor agonist binodenoson results in an extent and severity of reversible perfusion defects on SPECT imaging similar to nonselective adenosine receptor stimulation, accompanied by a dose-related reduction in the incidence and severity of side effects. PMID- 14734518 TI - Biological pacemaker implanted in canine left bundle branch provides ventricular escape rhythms that have physiologically acceptable rates. AB - BACKGROUND: We hypothesized that administration of the HCN2 gene to the left bundle-branch (LBB) system of intact dogs would provide pacemaker function in the physiological range of heart rates. METHODS AND RESULTS: An adenoviral construct incorporating HCN2 and green fluorescent protein (GFP) as a marker was injected via catheter under fluoroscopic control into the posterior division of the LBB. Controls were injected with an adenoviral construct of GFP alone or saline. Animals were monitored electrocardiographically for up to 7 days after surgery, at which time they were anesthetized and subjected to vagal stimulation to permit emergence of escape pacemakers. Hearts were then removed and injection sites visually identified and removed for microelectrode study of action potentials, patch clamp studies of pacemaker current, and/or immunohistochemical studies of HCN2. For 48 hours postoperatively, 7 of 7 animals subjected to 24-hour ECG monitoring showed multiple ventricular premature depolarizations and/or ventricular tachycardia attributable to injection-induced injury. Thereafter, sinus rhythm prevailed. During vagal stimulation, HCN2-injected dogs showed rhythms originating from the left ventricle, the rate of which was significantly more rapid than in the controls. Excised posterior divisions of the LBB from HCN2 injected animals manifested automatic rates significantly greater than the controls. Isolated tissues showed immunohistochemical and biophysical evidence of overexpressed HCN2. CONCLUSIONS: A gene-therapy approach for induction of biological pacemaker activity within the LBB system provides ventricular escape rhythms that have physiologically acceptable rates. Long-term stability and feasibility of the approach remain to be tested. PMID- 14734519 TI - Should surgeons take a break after an intraoperative death? Attitude survey and outcome evaluation. AB - OBJECTIVES: To investigate attitudes of cardiac surgeons and anaesthetists towards working immediately after an intraoperative death and to establish whether an intraoperative death affects the outcome of subsequent surgery. DESIGN: Questionnaire on attitudes to working after an intraoperative death and matched cohort study. SETTING: UK adult cardiac surgery centres and regional cardiothoracic surgical centre. PARTICIPANTS: 371 consultant cardiac surgeons and anaesthetists in the United Kingdom were asked to complete a questionnaire, and seven surgeons from one centre who continued to operate after intraoperative death. MAIN OUTCOME MEASURES: Outcome for 233 patients operated on by a surgeon who had experienced an intraoperative death within the preceding 48 hours compared with outcome of 932 matched controls. Hospital mortality and length of stay as a surrogate for hospital morbidity. RESULTS: The questionnaire response rate was 76%. Around a quarter of surgeons and anaesthetists thought they should stop work after an intraoperative death and most wanted guidelines on this subject. Overall, there was no increased mortality in patients operated on in the 48 hours after an intraoperative death. However, mortality was higher if the preceding intraoperative death was in an emergency or high risk case. Survivors operated on within 48 hours after an intraoperative death had longer stay in intensive care (odds ratio 1.64, 95% confidence interval 1.08 to 2.52, P = 0.02) and longer stay in hospital (relative change 1.15, 1.03 to 1.24, P = 0.02). CONCLUSION: Mortality is not increased in operations performed in the immediate aftermath of an intraoperative death, but survivors have longer stays in intensive care and on the hospital ward. PMID- 14734520 TI - Presentation of a self-peptide in two distinct conformations by a disease associated HLA-B27 subtype. PMID- 14734521 TI - Medullary epithelial cells of the human thymus express a highly diverse selection of tissue-specific genes colocalized in chromosomal clusters. AB - Promiscuous expression of tissue-specific self-antigens in the thymus imposes T cell tolerance and protects from autoimmune diseases, as shown in animal studies. Analysis of promiscuous gene expression in purified stromal cells of the human thymus at the single and global gene level documents the species conservation of this phenomenon. Medullary thymic epithelial cells overexpress a highly diverse set of genes (>400) including many tissue-specific antigens, disease-associated autoantigens, and cancer-germline genes. Although there are no apparent structural or functional commonalities among these genes and their products, they cluster along chromosomes. These findings have implications for human autoimmune diseases, immuno-therapy of tumors, and the understanding of the nature of this unorthodox regulation of gene expression. PMID- 14734522 TI - AIRE functions as an E3 ubiquitin ligase. AB - Autoimmune regulator (AIRE) gene mutation is responsible for the development of autoimmune-polyendocrinopathy-candidiasis ectodermal dystrophy, an organ-specific autoimmune disease with monogenic autosomal recessive inheritance. AIRE is predominantly expressed in medullary epithelial cells of the thymus and is considered to play important roles in the establishment of self-tolerance. AIRE contains two plant homeodomain (PHD) domains, and the novel role of PHD as an E3 ubiquitin (Ub) ligase has just emerged. Here we show that the first PHD (PHD1) of AIRE mediates E3 ligase activity. The significance of this finding was underscored by the fact that disease-causing missense mutations in the PHD1 (C311Y and P326Q) abolished its E3 ligase activity. These results add a novel enzymatic function for AIRE and suggest an indispensable role of the Ub proteasome pathway in the establishment of self-tolerance, in which AIRE is involved. PMID- 14734523 TI - Activation of arterial wall dendritic cells and breakdown of self-tolerance in giant cell arteritis. AB - Giant cell arteritis (GCA) is a granulomatous and occlusive vasculitis that causes blindness, stroke, and aortic aneurysm. CD4(+) T cells are selectively activated in the adventitia of affected arteries. In human GCA artery-severe combined immunodeficiency (SCID) mouse chimeras, depletion of CD83(+) dendritic cells (DCs) abrogated vasculitis, suggesting that DCs are critical antigen presenting cells in GCA. Healthy medium-size arteries possessed an indigenous population of DCs at the adventitia-media border. Adoptive T cell transfer into temporal artery-SCID mouse chimeras demonstrated that DCs in healthy arteries were functionally immature, but gained T cell stimulatory capacity after injection of lipopolysaccharide. In patients with polymyalgia rheumatica (PMR), a subclinical variant of GCA, adventitial DCs were mature and produced the chemokines CCL19 and CCL21, but vasculitic infiltrates were lacking. Human histocompatibility leukocyte antigen class II-matched healthy arteries, PMR arteries, and GCA arteries were coimplanted into SCID mice. Immature DCs in healthy arteries failed to stimulate T cells, but DCs in PMR arteries could attract, retain, and activate T cells that originated from the GCA lesions. We propose that in situ maturation of DCs in the adventitia is an early event in the pathogenesis of GCA. Activation of adventitial DCs initiates and maintains T cell responses in the artery and breaks tissue tolerance in the perivascular space. PMID- 14734524 TI - The activation status of neuroantigen-specific T cells in the target organ determines the clinical outcome of autoimmune encephalomyelitis. AB - The clinical picture of experimental autoimmune encephalomyelitis (EAE) is critically dependent on the nature of the target autoantigen and the genetic background of the experimental animals. Potentially lethal EAE is mediated by myelin basic protein (MBP)-specific T cells in Lewis rats, whereas transfer of S100beta- or myelin oligodendrocyte glycoprotein (MOG)-specific T cells causes intense inflammatory response in the central nervous system (CNS) with minimal disease. However, in Dark Agouti rats, the pathogenicity of MOG-specific T cells resembles the one of MBP-specific T cells in the Lewis rat. Using retrovirally transduced green fluorescent T cells, we now report that differential disease activity reflects different levels of autoreactive effector T cell activation in their target tissue. Irrespective of their pathogenicity, the migratory activity, gene expression patterns, and immigration of green fluorescent protein(+) T cells into the CNS were similar. However, exclusively highly pathogenic T cells were significantly reactivated within the CNS. Without local effector T cell activation, production of monocyte chemoattractants was insufficient to initiate and propagate a full inflammatory response. Low-level reactivation of weakly pathogenic T cells was not due to anergy because these cells could be activated by specific antigen in situ as well as after isolation ex vivo. PMID- 14734525 TI - Salmonella typhimurium persists within macrophages in the mesenteric lymph nodes of chronically infected Nramp1+/+ mice and can be reactivated by IFNgamma neutralization. AB - Host-adapted strains of Salmonella are capable of establishing a persistent infection in their host often in the absence of clinical disease. The mouse model of Salmonella infection has primarily been used as a model for the acute systemic disease. Therefore, the sites of long-term S. typhimurium persistence in the mouse are not known nor are the mechanisms of persistent infection clearly understood. Here, we show that S. typhimurium can persist for as long as 1 yr in the mesenteric lymph nodes (MLNs) of 129sv Nramp1(+)(/)(+) (Slc11a1(+)(/)(+)) mice despite the presence of high levels of anti-S. typhimurium antibody. Tissues from 129sv mice colonized for 60 d contain numerous inflammatory foci and lesions with features resembling S. typhi granulomas. Tissues from mice infected for 365 d have very few organized inflammatory lesions, but the bacteria continue to persist within macrophages in the MLN and the animals generally remain disease free. Finally, chronically infected mice treated with an interferon-gamma neutralizing antibody exhibited symptoms of acute systemic infection, with evidence of high levels of bacterial replication in most tissues and high levels of fecal shedding. Thus, interferon-gamma, which may affect the level of macrophage activation, plays an essential role in the control of the persistent S. typhimurium infection in mice. PMID- 14734526 TI - DNA polymerase eta is involved in hypermutation occurring during immunoglobulin class switch recombination. AB - Base substitutions, deletions, and duplications are observed at the immunoglobulin locus in DNA sequences involved in class switch recombination (CSR). These mutations are dependent upon activation-induced cytidine deaminase (AID) and present all the characteristics of the ones observed during V gene somatic hypermutation, implying that they could be generated by the same mutational complex. It has been proposed, based on the V gene mutation pattern of patients with the cancer-prone xeroderma pigmentosum variant (XP-V) syndrome who are deficient in DNA polymerase eta (pol eta), that this enzyme could be responsible for a large part of the mutations occurring on A/T bases. Here we show, by analyzing switched memory B cells from two XP-V patients, that pol eta is also an A/T mutator during CSR, in both the switch region of tandem repeats as well as upstream of it, thus suggesting that the same error-prone translesional polymerases are involved, together with AID, in both processes. PMID- 14734527 TI - Dual, HLA-B27 subtype-dependent conformation of a self-peptide. AB - The products of the human leukocyte antigen subtypes HLA-B*2705 and HLA-B*2709 differ only in residue 116 (Asp vs. His) within the peptide binding groove but are differentially associated with the autoimmune disease ankylosing spondylitis (AS); HLA-B*2705 occurs in AS-patients, whereas HLA-B*2709 does not. The subtypes also generate differential T cell repertoires as exemplified by distinct T cell responses against the self-peptide pVIPR (RRKWRRWHL). The crystal structures described here show that pVIPR binds in an unprecedented dual conformation only to HLA-B*2705 molecules. In one binding mode, peptide pArg5 forms a salt bridge to Asp116, connected with drastically different interactions between peptide and heavy chain, contrasting with the second, conventional conformation, which is exclusively found in the case of B*2709. These subtype-dependent differences in pVIPR binding link the emergence of dissimilar T cell repertoires in individuals with HLA-B*2705 or HLA-B*2709 to the buried Asp116/His116 polymorphism and provide novel insights into peptide presentation by major histocompatibility antigens. PMID- 14734528 TI - HIV-1 cell to cell transfer across an Env-induced, actin-dependent synapse. AB - Direct cell-cell transfer is an efficient mechanism of viral dissemination within an infected host, and human immunodeficiency virus 1 (HIV-1) can exploit this mode of spread. Receptor recognition by HIV-1 occurs via interactions between the viral surface envelope glycoprotein (Env), gp120, and CD4 and a chemokine receptor, CCR5 or CXCR4. Here, we demonstrate that the binding of CXCR4-using HIV 1-infected effector T cells to primary CD4(+)/CXCR4(+) target T cells results in rapid recruitment to the interface of CD4, CXCR4, talin, and lymphocyte function associated antigen 1 on the target cell, and of Env and Gag on the effector cell. Recruitment of these membrane molecules into polarized clusters was dependent on Env engagement of CD4 and CXCR4 and required remodelling of the actin cytoskeleton. Transfer of Gag from effector to target cell was observed by 1 h after conjugate formation, was independent of cell-cell fusion, and was probably mediated by directed virion fusion with the target cell. We propose that receptor engagement by Env directs the rapid, actin-dependent recruitment of HIV receptors and adhesion molecules to the interface, resulting in a stable adhesive junction across which HIV infects the target cell. PMID- 14734529 TI - Cell-to-cell transport of proteins and fluorescent tracers via plasmodesmata during plant development. AB - Plant cells communicate with each other via channels called plasmodesmata (PD). PD are not passive channels, but critical players in gene regulation, controlling intercellular transport of macromolecules between particular cells during development. PMID- 14734530 TI - FoxO3a regulates erythroid differentiation and induces BTG1, an activator of protein arginine methyl transferase 1. AB - Erythropoiesis requires tight control of expansion, maturation, and survival of erythroid progenitors. Because activation of phosphatidylinositol-3-kinase (PI3K) is required for erythropoietin/stem cell factor-induced expansion of erythroid progenitors, we examined the role of the PI3K-controlled Forkhead box, class O (FoxO) subfamily of Forkhead transcription factors. FoxO3a expression and nuclear accumulation increased during erythroid differentiation, whereas untimely induction of FoxO3a activity accelerated differentiation of erythroid progenitors to erythrocytes. We identified B cell translocation gene 1 (BTG1)/antiproliferative protein 2 as a FoxO3a target gene in erythroid progenitors. Promoter studies indicated BTG1 as a direct target of FoxO3a. Expression of BTG1 in primary mouse bone marrow cells blocked the outgrowth of erythroid colonies, which required a domain of BTG1 that binds protein arginine methyl transferase 1. During erythroid differentiation, increased arginine methylation coincided with BTG1 expression. Concordantly, inhibition of methyl transferase activity blocked erythroid maturation without affecting expansion of progenitor cells. We propose FoxO3a-controlled expression of BTG1 and subsequent regulation of protein arginine methyl transferase activity as a novel mechanism controlling erythroid expansion and differentiation. PMID- 14734531 TI - Trans-SNARE interactions elicit Ca2+ efflux from the yeast vacuole lumen. AB - Ca2+ transients trigger many SNARE-dependent membrane fusion events. The homotypic fusion of yeast vacuoles occurs after a release of lumenal Ca2+. Here, we show that trans-SNARE interactions promote the release of Ca2+ from the vacuole lumen. Ypt7p-GTP, the Sec1p/Munc18-protein Vps33p, and Rho GTPases, all of which function during docking, are required for Ca2+ release. Inhibitors of SNARE function prevent Ca2+ release. Recombinant Vam7p, a soluble Q-SNARE, stimulates Ca2+ release. Vacuoles lacking either of two complementary SNAREs, Vam3p or Nyv1p, fail to release Ca2+ upon tethering. Mixing these two vacuole populations together allows Vam3p and Nyv1p to interact in trans and rescues Ca2+ release. Sec17/18p promote sustained Ca2+ release by recycling SNAREs (and perhaps other limiting factors), but are not required at the release step itself. We conclude that trans-SNARE assembly events during docking promote Ca2+ release from the vacuole lumen. PMID- 14734532 TI - Regulation of polarized growth initiation and termination cycles by the polarisome and Cdc42 regulators. AB - The dynamic regulation of polarized cell growth allows cells to form structures of defined size and shape. We have studied the regulation of polarized growth using mating yeast as a model. Haploid yeast cells treated with high concentration of pheromone form successive mating projections that initiate and terminate growth with regular periodicity. The mechanisms that control the frequency of growth initiation and termination under these conditions are not well understood. We found that the polarisome components Spa2, Pea2, and Bni1 and the Cdc42 regulators Cdc24 and Bem3 control the timing and frequency of projection formation. Loss of polarisome components and mutation of Cdc24 decrease the frequency of projection formation, while loss of Bem3 increases the frequency of projection formation. We found that polarisome components and the cell fusion proteins Fus1 and Fus2 are important for the termination of projection growth. Our results define the first molecular regulators that control the timing of growth initiation and termination during eukaryotic cell differentiation. PMID- 14734533 TI - Novel regulation of mitotic exit by the Cdc42 effectors Gic1 and Gic2. AB - The guanine nucleotide exchange factor Cdc24, the GTPase Cdc42, and the Cdc42 effectors Cla4 and Ste20, two p21-activated kinases, form a signal transduction cascade that promotes mitotic exit in yeast. We performed a genetic screen to identify components of this pathway. Two related bud cortex-associated Cdc42 effectors, Gic1 and Gic2, were obtained as factors that promoted mitotic exit independently of Ste20. The mitotic exit function of Gic1 was dependent on its activation by Cdc42 and on the release of Gic1 from the bud cortex. Gic proteins became essential for mitotic exit when activation of the mitotic exit network through Cdc5 polo kinase and the bud cortex protein Lte1 was impaired. The mitotic exit defect of cdc5-10 Deltalte1 Deltagic1 Deltagic2 cells was rescued by inactivation of the inhibiting Bfa1-Bub2 GTPase-activating protein. Moreover, Gic1 bound directly to Bub2 and prevented binding of the GTPase Tem1 to Bub2. We propose that in anaphase the Cdc42-regulated Gic proteins trigger mitotic exit by interfering with Bfa1-Bub2 GTPase-activating protein function. PMID- 14734534 TI - Ordered proteolysis in anaphase inactivates Plk1 to contribute to proper mitotic exit in human cells. AB - We have found that key mitotic regulators show distinct patterns of degradation during exit from mitosis in human cells. Using a live-cell assay for proteolysis, we show that two of these regulators, polo-like kinase 1 (Plk1) and Aurora A, are degraded at different times after the anaphase-promoting complex/cyclosome (APC/C) switches from binding Cdc20 to Cdh1. Therefore, events in addition to the switch from Cdc20 to Cdh1 control the proteolysis of APC/C(Cdh1) substrates in vivo. We have identified a putative destruction box in Plk1 that is required for degradation of Plk1 in anaphase, and have examined the effect of nondegradable Plk1 on mitotic exit. Our results show that Plk1 proteolysis contributes to the inactivation of Plk1 in anaphase, and that this is required for the proper control of mitotic exit and cytokinesis. Our experiments reveal a role for APC/C mediated proteolysis in exit from mitosis in human cells. PMID- 14734535 TI - A divergent canonical WNT-signaling pathway regulates microtubule dynamics: dishevelled signals locally to stabilize microtubules. AB - Dishevelled (DVL) is associated with axonal microtubules and regulates microtubule stability through the inhibition of the serine/threonine kinase, glycogen synthase kinase 3beta (GSK-3beta). In the canonical WNT pathway, the negative regulator Axin forms a complex with beta-catenin and GSK-3beta, resulting in beta-catenin degradation. Inhibition of GSK-3beta by DVL increases beta-catenin stability and TCF transcriptional activation. Here, we show that Axin associates with microtubules and unexpectedly stabilizes microtubules through DVL. In turn, DVL stabilizes microtubules by inhibiting GSK-3beta through a transcription- and beta-catenin-independent pathway. More importantly, axonal microtubules are stabilized after DVL localizes to axons. Increased microtubule stability is correlated with a decrease in GSK-3beta-mediated phosphorylation of MAP-1B. We propose a model in which Axin, through DVL, stabilizes microtubules by inhibiting a pool of GSK-3beta, resulting in local changes in the phosphorylation of cellular targets. Our data indicate a bifurcation in the so-called canonical WNT-signaling pathway to regulate microtubule stability. PMID- 14734536 TI - A conserved protein interaction network involving the yeast MAP kinases Fus3 and Kss1. AB - The Saccharomyces cerevisiae mitogen-activated protein kinases (MAPKs) Fus3 and Kss1 bind to multiple regulators and substrates. We show that mutations in a conserved docking site in these MAPKs (the CD/7m region) disrupt binding to an important subset of their binding partners, including the Ste7 MAPK kinase, the Ste5 adaptor/scaffold protein, and the Dig1 and Dig2 transcriptional repressors. Supporting the possibility that Ste5 and Ste7 bind to the same region of the MAPKs, they partially competed for Fus3 binding. In vivo, some of the MAPK mutants displayed reduced Ste7-dependent phosphorylation, and all of them exhibited multiple defects in mating and pheromone response. The Kss1 mutants were also defective in Kss1-imposed repression of Ste12. We conclude that MAPKs contain a structurally and functionally conserved docking site that mediates an overall positively acting network of interactions with cognate docking sites on their regulators and substrates. Key features of this interaction network appear to have been conserved from yeast to humans. PMID- 14734538 TI - Physical and functional interaction of the active zone proteins, CAST, RIM1, and Bassoon, in neurotransmitter release. AB - We have recently isolated a novel cytomatrix at the active zone (CAZ)-associated protein, CAST, and found it directly binds another CAZ protein RIM1 and indirectly binds Munc13-1 through RIM1; RIM1 and Munc13-1 directly bind to each other and are implicated in priming of synaptic vesicles. Here, we show that all the CAZ proteins thus far known form a large molecular complex in the brain, including CAST, RIM1, Munc13-1, Bassoon, and Piccolo. RIM1 and Bassoon directly bind to the COOH terminus and central region of CAST, respectively, forming a ternary complex. Piccolo, which is structurally related to Bassoon, also binds to the Bassoon-binding region of CAST. Moreover, the microinjected RIM1- or Bassoon binding region of CAST impairs synaptic transmission in cultured superior cervical ganglion neurons. Furthermore, the CAST-binding domain of RIM1 or Bassoon also impairs synaptic transmission in the cultured neurons. These results indicate that CAST serves as a key component of the CAZ structure and is involved in neurotransmitter release by binding these CAZ proteins. PMID- 14734537 TI - Atypical protein kinase C (PKCzeta/lambda) is a convergent downstream target of the insulin-stimulated phosphatidylinositol 3-kinase and TC10 signaling pathways. AB - Insulin stimulation of adipocytes resulted in the recruitment of atypical PKC (PKCzeta/lambda) to plasma membrane lipid raft microdomains. This redistribution of PKCzeta/lambda was prevented by Clostridium difficile toxin B and by cholesterol depletion, but was unaffected by inhibition of phosphatidylinositol (PI) 3-kinase activity. Expression of the constitutively active GTP-bound form of TC10 (TC10Q/75L), but not the inactive GDP-bound mutant (TC10/T31N), targeted PKCzeta/lambda to the plasma membrane through an indirect association with the Par6-Par3 protein complex. In parallel, insulin stimulation as well as TC10/Q75L resulted in the activation loop phosphorylation of PKCzeta. Although PI 3-kinase activation also resulted in PKCzeta/lambda phosphorylation, it was not recruited to the plasma membrane. Furthermore, insulin-induced GSK-3beta phosphorylation was mediated by both PI 3-kinase-PKB and the TC10-Par6-atypical PKC signaling pathways. Together, these data demonstrate that PKCzeta/lambda can serve as a convergent downstream target for both the PI 3-kinase and TC10 signaling pathways, but only the TC10 pathway induces a spatially restricted targeting to the plasma membrane. PMID- 14734539 TI - Sinuous is a Drosophila claudin required for septate junction organization and epithelial tube size control. AB - Epithelial tubes of the correct size and shape are vital for the function of the lungs, kidneys, and vascular system, yet little is known about epithelial tube size regulation. Mutations in the Drosophila gene sinuous have previously been shown to cause tracheal tubes to be elongated and have diameter increases. Our genetic analysis using a sinuous null mutation suggests that sinuous functions in the same pathway as the septate junction genes neurexin and scribble, but that nervana 2, convoluted, varicose, and cystic have functions not shared by sinuous. Our molecular analyses reveal that sinuous encodes a claudin that localizes to septate junctions and is required for septate junction organization and paracellular barrier function. These results provide important evidence that the paracellular barriers formed by arthropod septate junctions and vertebrate tight junctions have a common molecular basis despite their otherwise different molecular compositions, morphologies, and subcellular localizations. PMID- 14734540 TI - Substitutions at the Asp-473 latch residue of chlamydomonas ribulosebisphosphate carboxylase/oxygenase cause decreases in carboxylation efficiency and CO(2)/O(2) specificity. AB - The loop between alpha-helix 6 and beta-strand 6 in the alpha/beta-barrel active site of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco, EC 4.1.1.39) plays a key role in discriminating between gaseous substrates CO(2) and O(2). Based on numerous x-ray crystal structures, loop 6 is either closed or open depending on the presence or absence, respectively, of substrate ligands. The carboxyl terminus folds over loop 6 in the closed conformation, prompting speculation that it may trigger or latch loop 6 closure. Because an x-ray crystal structure of tobacco Rubisco revealed that phosphate is located at a site in the open form that is occupied by the carboxyl group of Asp-473 in the closed form, it was proposed that Asp-473 may serve as the latch that holds the carboxyl terminus over loop 6. To assess the essentiality of Asp-473 in catalysis, we used directed mutagenesis and chloroplast transformation of the green alga Chlamydomonas reinhardtii to create D473A and D473E mutant enzymes. The D473A and D473E mutant strains can grow photoautotrophically, indicating that Asp-473 is not essential for catalysis. However, both substitutions caused 87% decreases in carboxylation catalytic efficiency (V(max)/K(m)) and approximately 16% decreases in CO(2)/O(2) specificity. If the carboxyl terminus is required for stabilizing loop 6 in the closed conformation, there must be additional residues at the carboxyl terminus/loop 6 interface that contribute to this mechanism. Considering that substitutions at residue 473 can influence CO(2)/O(2) specificity, further study of interactions between loop 6 and the carboxyl terminus may provide clues for engineering an improved Rubisco. PMID- 14734541 TI - pH dependence and stoichiometry of binding to the Fc region of IgG by the herpes simplex virus Fc receptor gE-gI. AB - Herpes simplex virus type 1 encodes two glycoproteins, gE and gI, that form a heterodimer on the surface of virions and infected cells. The gE-gI heterodimer has been implicated in cell-to-cell spread of virus and is a receptor for the Fc fragment of IgG. Previous studies localized the gE-gI-binding site on human IgG to a region near the interface between the C(H)2 and C(H)3 domains of Fc, which also serves as the binding site for bacterial and mammalian Fc receptors. Although there are two potential gE-gI-binding sites per Fc homodimer, only one gE-gI heterodimer binds per IgG in gel filtration experiments. Here we report production of recombinant human Fc molecules that contain zero, one, or two potential gE-gI-binding sites and use them in analytical ultracentrifugation experiments to show that two gE-gI heterodimers can bind to each Fc. Further characterization of the gE-gI interaction with Fc reveals a sharp pH dependence of binding, with K(D) values of approximately 340 and approximately 930 nm for the first and second binding events, respectively, at the slightly basic pH of the cell surface (pH 7.4), but undetectable binding at pH 6.0. This strongly pH dependent interaction suggests a physiological role for gE-gI dissociation from IgG within acidic intracellular compartments, consistent with a mechanism whereby herpes simplex virus promotes intracellular degradation of anti-viral antibodies. PMID- 14734542 TI - Monomeric Cu,Zn-superoxide dismutase is a common misfolding intermediate in the oxidation models of sporadic and familial amyotrophic lateral sclerosis. AB - Proteinacious intracellular aggregates in motor neurons are a key feature of both sporadic and familial amyotrophic lateral sclerosis (ALS). These inclusion bodies are often immunoreactive for Cu,Zn-superoxide dismutase (SOD1) and are implicated in the pathology of ALS. On the basis of this and a similar clinical presentation of symptoms in the familial (fALS) and sporadic forms of ALS, we sought to investigate the possibility that there exists a common disease-related aggregation pathway for fALS-associated mutant SODs and wild type SOD1. We have previously shown that oxidation of fALS-associated mutant SODs produces aggregates that have the same morphological, structural, and tinctorial features as those found in SOD1 inclusion bodies in ALS. Here, we show that oxidative damage of wild type SOD at physiological concentrations ( approximately 40 microm) results in destabilization and aggregation in vitro. Oxidation of either mutant or wild type SOD1 causes the enzyme to dissociate to monomers prior to aggregation. Only small changes in secondary and tertiary structure are associated with monomer formation. These results indicate a common aggregation prone monomeric intermediate for wild type and fALS-associated mutant SODs and provides a link between sporadic and familial ALS. PMID- 14734543 TI - Interactions and reactions of ferritin with DNA. AB - Ferritin, normally considered a cytoplasmic iron-storage protein, is also found in the nuclei of some cells. There is no current agreement about its function(s) in this environment. Proposals include DNA protection, provision of iron to nuclear enzymes, and regulation of transcription initiation, but evidence for these functions is scanty. We have shown previously that H-ferritin subunits can be cross-linked to chromosomal DNA in vivo (Thompson, K. J., Fried, M. G., Ye, Z., Boyer, P., and Connor, J. R. (2002) J. Cell Sci. 115, 2165-2177). Here we describe systematic analyses of DNA binding and the covalent stability of DNA in the presence of ferritins from several different sources. Our data show that the H-subunit of human ferritin binds DNA, whereas neither the L-subunit nor the ferroxidase-deficient 222-mutant of the H-subunit has detectable binding activity. DNA binding is without significant preference for base composition, sequence, or the nature of DNA ends. H- and L-ferritins and ferritins of mixed subunit composition stimulate the conversion of superhelical plasmid DNA to the relaxed form. The sensitivity of this conversion to glycerol suggests that DNA is nicked by a free radical mechanism. The rate of nicking correlates with the iron content of the ferritin and is strongly inhibited by chelators. Ferritin dependent nicking is characterized by a kinetic lag that is not seen in control reactions containing free iron species. These results suggest that the release of iron from ferritin is an important part of the nicking mechanism. The potential role of ferritin as a protector of the genome is discussed in the context of these results. PMID- 14734544 TI - A PBP2x from a clinical isolate of Streptococcus pneumoniae exhibits an alternative mechanism for reduction of susceptibility to beta-lactam antibiotics. AB - The human pathogen Streptococcus pneumoniae is one of the main causative agents of respiratory tract infections. At present, clinical isolates of S. pneumoniae often exhibit decreased susceptibility toward beta-lactams, a phenomenon linked to multiple mutations within the penicillin-binding proteins (PBPs). PBP2x, one of the six PBPs of S. pneumoniae, is the first target to be modified under antibiotic pressure. By comparing 89 S. pneumoniae PBP2x sequences from clinical and public data bases, we have identified one major group of sequences from drug sensitive strains as well as two distinct groups from drug-resistant strains. The first group includes proteins that display high similarity to PBP2x from the well characterized resistant strain Sp328. The second group includes sequences in which a signature mutation, Q552E, is found adjacent to the third catalytic motif. In this work, a PBP2x from a representative strain from the latter group (S. pneumoniae 5259) was biochemically and structurally characterized. Phenotypical analyses of transformed pneumococci show that the Q552E substitution is responsible for most of the reduction of strain susceptibility toward beta lactams. The crystal structure of 5259-PBP2x reveals a change in polarity and charge distribution around the active site cavity, as well as rearrangement of strand beta3, emulating structural changes observed for other PBPs that confer drug resistance to Gram-positive pathogens. Interestingly, the active site of 5259-PBP2x is in closed conformation, whereas that of Sp328-PBP2x is open. Consequently, S. pneumoniae has evolved to employ the same protein in two distinct mechanisms of antibiotic resistance. PMID- 14734545 TI - Substrate requirements of the oxygen-sensing asparaginyl hydroxylase factor inhibiting hypoxia-inducible factor. AB - The hypoxia-inducible factor alpha subunits 1 and 2 (HIF-1alpha and HIF-2alpha) are subjected to oxygen-dependent asparaginyl hydroxylation, a modification that represses the carboxyl-terminal transactivation domain (CAD) at normoxia by preventing recruitment of the p300/cAMP-response element-binding protein coactivators. This hydroxylation is performed by the novel asparaginyl hydroxylase, factor-inhibiting HIF-1' (FIH-1), of which HIF-1alpha and HIF-2alpha are the only reported substrates. Here we investigated the substrate requirements of FIH-1 by characterizing its subcellular localization and by examining amino acids within the HIF-1alpha substrate for their importance in recognition and catalysis by FIH-1. Using immunohistochemistry, we showed that both endogenous and transfected FIH-1 are primarily confined to the cytoplasm and remain there under normoxia and following treatment with the hypoxia mimetic, dipyridyl. Individual alanine mutations of seven conserved amino acids flanking the hydroxylated asparagine in HIF-1alpha revealed the importance of the valine (Val 802) adjacent to the targeted asparagine. The HIF-1alpha CAD V802A mutant exhibited a 4-fold lower V(max) in enzyme assays, whereas all other mutants were hydroxylated as efficiently as the wild type HIF-1alpha CAD. Furthermore, in cell based assays the transcriptional activity of V802A was constitutive, suggesting negligible normoxic hydroxylation in HEK293T cells, whereas the wild type and other mutants were repressed under normoxia. Molecular modeling of the HIF-1alpha CAD V802A in complex with FIH-1 predicted an alteration in asparagine positioning compared with the wild type HIF-1alpha CAD, providing an explanation for the impaired catalysis observed and confirming the importance of Val-802 in asparaginyl hydroxylation by FIH-1. PMID- 14734546 TI - Inositol deacylation of glycosylphosphatidylinositol-anchored proteins is mediated by mammalian PGAP1 and yeast Bst1p. AB - The inositol moiety of mammalian glycosylphosphatidylinositol (GPI) is acylated at an early step in GPI biosynthesis. The inositol acylation is essential for the generation of mature GPI capable of attachment to proteins. However, the acyl group is usually absent from GPI-anchored proteins (GPI-APs) on the cell surface due to inositol deacylation that occurs in the endoplasmic reticulum (ER) soon after GPI-anchor attachment. Mammalian GPI inositol-deacylase has not been cloned, and the biological significance of the deacylation has been unclear. Here we report a GPI inositol-deacylase-deficient Chinese hamster ovary cell line established by taking advantage of resistance to phosphatidylinositol-specific phospholipase C and the gene responsible, which was termed PGAP1 for Post GPI Attachment to Proteins 1. PGAP1 encoded an ER-associated, 922-amino acid membrane protein bearing a lipase consensus motif. Substitution of a conserved putative catalytic serine with alanine resulted in a complete loss of function, indicating that PGAP1 is the GPI inositol-deacylase. The mutant cells showed a clear delay in the maturation of GPI-APs in the Golgi and accumulation of GPI-APs in the ER. Thus, the GPI inositol deacylation is important for efficient transport of GPI APs from the ER to the Golgi. PMID- 14734547 TI - Physical and functional interaction between the XPF/ERCC1 endonuclease and hRad52. AB - The XPF/ERCC1 heterodimer is a DNA structure-specific endonuclease that participates in nucleotide excision repair and homology-dependent recombination reactions, including DNA single strand annealing and gene targeting. Here we show that XPF/ERCC1 is stably associated with hRad52, a recombinational repair protein, in human cell-free extracts and that these factors interact directly via the N-terminal domain of hRad52 and the XPF protein. Complex formation between hRad52 and XPF/ERCC1 concomitantly stimulates the DNA structure-specific endonuclease activity of XPF/ERCC1 and attenuates the DNA strand annealing activity of hRad52. Our results reveal a novel role for hRad52 as a subunit of a DNA structure-specific endonuclease and are congruent with evidence implicating both hRad52 and XPF/ERCC1 in a number of homologous recombination reactions. We propose that the ternary complex of hRad52 and XPF/ERCC1 is the active species that processes recombination intermediates generated during the repair of DNA double strand breaks and in homology-dependent gene targeting events. PMID- 14734548 TI - Distant structural homology leads to the functional characterization of an archaeal PIN domain as an exonuclease. AB - Genome sequencing projects have focused attention on the problem of discovering the functions of protein domains that are widely distributed throughout living species but which are, as yet, largely uncharacterized. One such example is the PIN domain, found in eukaryotes, bacteria, and Archaea, and with suggested roles in signaling, RNase editing, and/or nucleotide binding. The first reported crystal structure of a PIN domain (open reading frame PAE2754, derived from the crenarchaeon, Pyrobaculum aerophilum) has been determined to 2.5 A resolution and is presented here. Mapping conserved residues from a multiple sequence alignment onto the structure identifies a putative active site. The discovery of distant structural homology with several exonucleases, including T4 phage RNase H and flap endonuclease (FEN1), further suggests a likely function for PIN domains as Mg2+-dependent exonucleases, a hypothesis that we have confirmed in vitro. The tetrameric structure of PAE2754, with the active sites inside a tunnel, suggests a mechanism for selective cleavage of single-stranded overhangs or flap structures. These results indicate likely DNA or RNA editing roles for prokaryotic PIN domains, which are strikingly numerous in thermophiles, and in organisms such as Mycobacterium tuberculosis. They also support previous hypotheses that eukaryotic PIN domains participate in RNAi and nonsense-mediated RNA degradation. PMID- 14734549 TI - Controlling the substrate selectivity of deacetoxycephalosporin/deacetylcephalosporin C synthase. AB - Deacetoxycephalosporin/deacetylcephalosporin C synthase (DAOC/DACS) is an iron(II) and 2-oxoglutarate-dependent oxygenase involved in the biosynthesis of cephalosporin C in Cephalosporium acremonium. It catalyzes two oxidative reactions, oxidative ring-expansion of penicillin N to deacetoxycephalosporin C, and hydroxylation of the latter to give deacetylcephalosporin C. The enzyme is closely related to deacetoxycephalosporin C synthase (DAOCS) and DACS from Streptomyces clavuligerus, which selectively catalyze ring-expansion or hydroxylation reactions, respectively. In this study, structural models based on DAOCS coupled with site-directed mutagenesis were used to identify residues within DAOC/DACS that are responsible for controlling substrate and reaction selectivity. The M306I mutation abolished hydroxylation of deacetylcephalosporin C, whereas the W82A mutant reduced ring-expansion of penicillin G (an "unnatural" substrate). Truncation of the C terminus of DAOC/DACS to residue 310 (Delta310 mutant) enhanced ring-expansion of penicillin G by approximately 2-fold. A double mutant, Delta310/M306I, selectively catalyzed the ring-expansion reaction and had similar kinetic parameters to the wild-type DAOC/DACS. The Delta310/N305L/M306I triple mutant selectively catalyzed ring-expansion of penicillin G and had improved kinetic parameters (K(m) = 2.00 +/- 0.47 compared with 6.02 +/- 0.97 mm for the wild-type enzyme). This work demonstrates that a single amino acid residue side chain within the DAOC/DACS active site can control whether the enzyme catalyzes ring-expansion, hydroxylation, or both reactions. The catalytic efficiency of mutant enzymes can be improved by combining active site mutations with other modifications including C-terminal truncation and modification of Asn 305. PMID- 14734550 TI - Identification of two RNA cis-elements that function to regulate the 5' splice site selection of Bcl-x pre-mRNA in response to ceramide. AB - Two splice variants derived from the BCL-x gene, proapoptotic Bcl-x(s) and anti apoptotic Bcl-x(L), are produced via alternative 5' splice site selection within exon 2 of Bcl-x pre-mRNA. In previous studies, our laboratory demonstrated that ceramide regulated this 5' splice site selection, inducing the production of Bcl x(s) mRNA with a concomitant decrease in Bcl-x(L) correlating with sensitization to chemotherapy (Chalfant, C. E., Rathman, K., Pinkerman, R. L., Wood, R. E., Obeid, L. M., Ogretmen, B., and Hannun, Y. A. (2002) J. Biol. Chem. 277, 12587 12595). We have now identified several possible RNA cis-elements within exon 2 of Bcl-x pre-mRNA by sequence analysis. To study the possible roles of these RNA cis elements in regulating the alternative 5' splice site selection of Bcl-x pre mRNA, we developed a BCL-x minigene construct which conferred the same ratio of Bcl-x(L)/Bcl-x(s) mRNA as the endogenous Bcl-x and was responsive to ceramide treatment. Mutagenesis of either a purine-rich splicing enhancer or a pyrimidine tract element within exon 2 induced a change in the ratio of Bcl-x(L)/Bcl-x(s) mRNA from 7 to 1 and 0.23, thereby diminishing the selection of the Bcl-x(L) 5' splice site with a concomitant increase in Bcl-x(s) 5' splice site selection. Furthermore, mutagenesis of these cis-elements abolished the ability of ceramide to affect the 5' splice site selection. In vitro binding assays coupled with competitor studies demonstrated specific binding of RNA trans-activating proteins to these regions. SDS-PAGE analysis of cross-linked RNA trans-activating factors with these RNA cis-elements revealed the binding of 215-, 120-, and 30-kDa proteins to the purine-rich element and 120- and 76-kDa proteins to the pyrimidine tract element. In addition, exogenous treatment of A549 cells with ceramide increased the formation of protein complexes with these RNA cis elements. Therefore, we have identified two ceramide-responsive RNA cis-elements within exon 2 of Bcl-x pre-mRNA, and this is the first report of an RNA cis element responsive to a bioactive lipid. PMID- 14734551 TI - Interleukin (IL)-1F6, IL-1F8, and IL-1F9 signal through IL-1Rrp2 and IL-1RAcP to activate the pathway leading to NF-kappaB and MAPKs. AB - Interleukin 1 (IL-1) plays a prominent role in immune and inflammatory reactions. Our understanding of the IL-1 family has recently expanded to include six novel members named IL-1F5 to IL-1F10. Recently, it was reported that IL-1F9 activated NF-kappaB through the orphan receptor IL-1 receptor (IL-1R)-related protein 2 (IL 1Rrp2) in Jurkat cells (Debets, R., Timans, J. C., Homey, B., Zurawski, S., Sana, T. R., Lo, S., Wagner, J., Edwards, G., Clifford, T., Menon, S., Bazan, J. F., and Kastelein, R. A. (2001) J. Immunol. 167, 1440-1446). In this study, we demonstrate that IL-1F6 and IL-1F8, in addition to IL-1F9, activate the pathway leading to NF-kappaB in an IL-1Rrp2-dependent manner in Jurkat cells as well as in multiple other human and mouse cell lines. Activation of the pathway leading to NF-kappaB by IL-1F6 and IL-1F8 follows a similar time course to activation by IL-1beta, suggesting that signaling by the novel family members occurs through a direct mechanism. In a mammary epithelial cell line, NCI/ADR-RES, which naturally expresses IL-1Rrp2, all three cytokines signal without further receptor transfection. IL-1Rrp2 antibodies block activation of the pathway leading to NF kappaB by IL-1F6, IL-1F8, and IL-1F9 in both Jurkat and NCI/ADR-RES cells. In NCI/ADR-RES cells, the three IL-1 homologs activated the MAPKs, JNK and ERK1/2, and activated downstream targets as well, including an IL-8 promoter reporter and the secretion of IL-6. We also provide evidence that IL-1RAcP, in addition to IL 1Rrp2, is required for signaling by all three cytokines. Antibodies directed against IL-1RAcP and transfection of cytoplasmically deleted IL-1RAcP both blocked activation of the pathway leading to NF-kappaB by the three cytokines. We conclude that IL-1F6, IL-1F8, and IL-1F9 signal through IL-1Rrp2 and IL-1RAcP. PMID- 14734552 TI - A novel missense mutation in AE1 causing autosomal dominant distal renal tubular acidosis retains normal transport function but is mistargeted in polarized epithelial cells. AB - Mutations in SLC4A1, encoding the chloride-bicarbonate exchanger AE1, cause distal renal tubular acidosis (dRTA), a disease of defective urinary acidification by the distal nephron. In this study we report a novel missense mutation, G609R, causing dominant dRTA in affected members of a large Caucasian pedigree who all exhibited metabolic acidosis with alkaline urine, prominent nephrocalcinosis, and progressive renal impairment. To investigate the potential disease mechanism, the consequent effects of this mutation were determined. We first assessed anion transport function of G609R by expression in Xenopus oocytes. Western blotting and immunofluorescence demonstrated that the mutant protein was expressed at the oocyte cell surface. Measuring chloride and bicarbonate fluxes revealed normal 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid-inhibitable anion exchange, suggesting that loss-of-function of kAE1 cannot explain the severe disease phenotype in this kindred. We next expressed epitope tagged wild-type or mutant kAE1 in Madin-Darby canine kidney cells. In monolayers grown to polarity, mutant kAE1 was detected subapically and at the apical membrane, as well as at the basolateral membrane, in contrast to the normal basolateral appearance of wild-type kAE1. These findings suggest that the seventh transmembrane domain that contains Gly-609 plays an important role in targeting kAE1 to the correct cell surface compartment. They confirm that dominant dRTA is associated with non-polarized trafficking of the protein, with no significant effect on anion transport function in vitro, which remains an unusual mechanism of human disease. PMID- 14734553 TI - Attenuation of HIV-1 replication in primary human cells with a designed zinc finger transcription factor. AB - Small molecule inhibitors of human immunodeficiency virus, type 1 (HIV-1) have been extremely successful but are associated with a myriad of undesirable effects and require lifelong daily dosing. In this study we explore an alternative approach, that of inducing intracellular immunity using designed, zinc finger based transcription factors. Three transcriptional repression proteins were engineered to bind sites in the HIV-1 promoter that were expected to be both accessible in chromatin structure and highly conserved in sequence structure among the various HIV-1 subgroups. Transient transfection assays identified one factor, KRAB-HLTR3, as being able to achieve 100-fold repression of an HIV-1 promoter. Specificity of repression was demonstrated by the lack of repression of other promoters. This factor was further shown to repress the replication of several HIV-1 viral strains 10- to 100-fold in T-cell lines and primary human peripheral blood mononuclear cells. Repression was observed for at least 18 days with no significant cytotoxicity. Stable T-cell lines expressing the factor also do not show obvious signs of cytotoxicity. These characteristics present KRAB HLTR3 as an attractive candidate for development in an intracellular immunization strategy for anti-HIV-1 therapy. PMID- 14734554 TI - Differential specificity of human and Escherichia coli endonuclease III and VIII homologues for oxidative base lesions. AB - In human cells, oxidative pyrimidine lesions are restored by the base excision repair pathway initiated by homologues of Endo III (hNTH1) and Endo VIII (hNEIL1 and hNEIL2). In this study we have quantitatively analyzed and compared their activity toward nine oxidative base lesions and an apurinic/apyrimidinic (AP) site using defined oligonucleotide substrates. hNTH1 and hNEIL1 but not hNEIL2 excised the two stereoisomers of thymine glycol (5R-Tg and 5S-Tg), but their isomer specificity was markedly different: the relative activity for 5R-Tg:5S-Tg was 13:1 for hNTH1 and 1.5:1 for hNEIL1. This was also the case for their Escherichia coli homologues: the relative activity for 5R-Tg:5S-Tg was 1:2.5 for Endo III and 3.2:1 for Endo VIII. Among other tested lesions for hNTH1, an AP site was a significantly better substrate than urea, 5-hydroxyuracil (hoU), and guanine-derived formamidopyrimidine (mFapyG), whereas for hNEIL1 these base lesions and an AP site were comparable substrates. In contrast, hNEIL2 recognized an AP site exclusively, and the activity for hoU and mFapyG was marginal. hNEIL1, hNEIL2, and Endo VIII but not hNTH1 and Endo III formed cross-links to oxanine, suggesting conservation of the -fold of the active site of the Endo VIII homologues. The profiles of the excision of the Tg isomers with HeLa and E. coli cell extracts closely resembled those of hNTH1 and Endo III, confirming their major contribution to the repair of Tg isomers in cells. However, detailed analysis of the cellular activity suggests that hNEIL1 has a significant role in the repair of 5S-Tg in human cells. PMID- 14734555 TI - Systematic mutagenesis of the leucine-rich repeat (LRR) domain of CCR4 reveals specific sites for binding to CAF1 and a separate critical role for the LRR in CCR4 deadenylase activity. AB - CCR4, a poly(A) deadenylase of the exonuclease III family, is a component of the multiprotein CCR4-NOT complex of Saccharomyces cerevisiae that is involved in mRNA degradation. CCR4, unlike all other exonuclease III family members, contains a leucine-rich repeat (LRR) motif through which it makes contact to CAF1 and other factors. The LRR residues important in contacting CAF1 were identified by constructing 29 CCR4 mutations encompassing a majority (47 of 81) of residues interstitial to the conserved structural residues. Two-hybrid and immunoprecipitation data revealed that physical contact between CAF1 and the LRR is blocked by mutation of just two alpha-helix/beta-helix strand loop residues linking the first and second repeats. In contrast, CAF16, a potential ligand of CCR4, was abrogated in its binding to the LRR by mutations in the N terminus of the second beta-strand. The LRR domain was also found to contact the deadenylase domain of CCR4, and deletion of the LRR region completely inhibited CCR4 enzymatic activity. Mutations throughout the beta-sheet surface of the LRR, including those that did not specifically interfere with contacts to CAF1 or CAF16, significantly reduced CCR4 deadenylase activity. These results indicate that the CCR4-LRR, in addition to binding to CAF1, plays an essential role in the CCR4 deadenylation of mRNA. PMID- 14734556 TI - RGS6 interacts with DMAP1 and DNMT1 and inhibits DMAP1 transcriptional repressor activity. AB - RGS6 is a member of a subfamily of mammalian RGS proteins that possess DEP (disheveled, Egl-10, pleckstrin) and GGL (G protein gamma subunit-like) domains in addition to the hallmark RGS domain. RGS proteins negatively regulate heterotrimeric G protein signaling by virtue of the GTPase-activating protein activity of their RGS domains. RGS6 exists in multiple splice forms with a long (6L) or short (6S) N terminus, a complete or incomplete GGL domain, in combination with various C-terminal domains. Green fluorescent protein-tagged RGS6L and RGS6S forms exhibit predominantly cytoplasmic and nuclear patterns of distribution in COS-7 cells, respectively, and traffic from these sites to nucleoli in response to stress signaling. We undertook a yeast two-hybrid screen for nuclear RGS6-binding proteins and here identify DMAP1 as an RGS6-interacting protein. DMAP1 is a component of the Dnmt1 complex involved in repression of newly replicated genes. The domains of interaction were mapped to the N-terminal region of the GGL domain of RGS6, a region distinct from its Gbeta5 binding region, and the C-terminal domain of DMAP1. Gbeta5 and DMAP1 did not compete for each other's interaction with RGS6. Co-immunoprecipitation studies in COS-7 cells showed that RGS6L and RGS6S, but not RGS6LDelta258-293 deletion mutant lacking a DMAP1-binding module, co-immunoprecipitate DMAP1 as well as Dnmt1 in a DMAP1 dependent manner. A recombinant GGL domain of RGS6 precipitated endogenous DMAP1 and Dnmt1 in neuroblastoma cell lysates and endogenous DMAP1 co immunoprecipitated with RGS6L from mouse brain. Co-expression of DMAP1 with RGS6L promoted nuclear migration of RGS6L and its co-localization with DMAP1, a response not observed with RGS6LDelta258-293. RGS6 inhibited the transcriptional repressor activity of DMAP1. RGS6 is the first member of the RGS protein family shown to interact with proteins involved in transcriptional regulation. PMID- 14734557 TI - Effects of cholesterol depletion and increased lipid unsaturation on the properties of endocytic membranes. AB - Lipid analogs with dialkylindocarbocyanine (DiI) head groups and short or unsaturated hydrocarbon chains (e.g. DiIC(12) and FAST DiI) enter the endocytic recycling compartment efficiently, whereas lipid analogs with long, saturated tails (e.g. DiIC(16) and DiIC(18)) are sorted out of this pathway and targeted to the late endosomes/lysosomes (Mukherjee, S., Soe, T. T., and Maxfield, F. R. (1999) J. Cell Biol. 144, 1271-1284). This differential trafficking of lipid analogs with the same polar head group was interpreted to result from differential partitioning to different types of domains with varying membrane order and/or curvature. Here we investigate the system further by monitoring the trafficking behavior of these lipid analogs under conditions that alter domain properties. There was a marked effect of cholesterol depletion on the cell surface distribution and degree of internalization of the lipid probes. Furthermore, instead of going to the late endosomes/lysosomes as in control cells, long chain DiI analogs, such as DiIC(16), were sorted to the recycling pathway in cholesterol-depleted cells. We confirmed that this difference was due to a change in overall membrane properties, and not cholesterol levels per se, by utilizing a Chinese hamster ovary cell line that overexpressed transfected stearoyl-CoA desaturase 1, a rate-limiting enzyme in the production of monounsaturated fatty acids. These cells have a decrease in membrane order because they contain a much larger fraction of unsaturated fatty acids. These cells showed alteration of DiI trafficking very similar to cholesterol-depleted cells. By using cold Triton X-100 extractability of different lipids as a criterion to determine the membrane properties of intracellular organelles, we found that the endocytic recycling compartment has abundant detergent-resistant membranes, in contrast to the late endosomes and lysosomes. PMID- 14734558 TI - Silencing of SPC2 expression using an engineered delta ribozyme in the mouse betaTC-3 endocrine cell line. AB - Endoproteolytic processing is carried out by subtilase-like pro-protein convertases in mammalian cells. In order to understand the distinct roles of a member of this family (SPC2), gene silencing in cultured cells is an ideal approach. Previous studies showed limited success in either the degree of inhibition obtained or the stability of the cell lines. Here we demonstrate the high potential of delta ribozyme as a post-transcriptional gene silencing tool in cultured cells. We used an expression vector based on the RNA polymerase III promoter to establish betaTC-3 stable cell lines expressing the chimeric tRNA(Val)-delta ribozyme transcript targeting SPC2 mRNA. Northern and Western blot hybridizations showed a specific reduction of SPC2 mRNA and protein. Validation of processing effects was tested by measuring the levels of dynorphin A-(1-8), which are present in betaTC-3 cells as a result of the unique cleavage of dynorphin A-(1-17) by SPC2. Moreover, a differential proteomic analysis confirmed these results and allowed identification of secretogranin II as a potential substrate of SPC2. The development of efficient, specific, and durable silencing tools, such as described in the present work, will be of great importance in elucidating the functions of the subtilase-like pro-protein convertases in regard to peptide processing and derived cellular events. PMID- 14734559 TI - Specificity and enzyme kinetics of the quorum-quenching N-Acyl homoserine lactone lactonase (AHL-lactonase). AB - N-Acyl homoserine lactone (AHL) quorum-sensing signals are the vital elements of bacterial quorum-sensing systems, which regulate diverse biological functions, including virulence. The AHL-lactonase, a quorumquenching enzyme encoded by aiiA from Bacillus sp., inactivates AHLs by hydrolyzing the lactone bond to produce corresponding N-acyl homoserines. To characterize the enzyme, the recombinant AHL lactonase and its four variants were purified. Kinetic and substrate specificity analysis showed that AHL-lactonase had no or little residue activity to non-acyl lactones and noncyclic esters, but displayed strong enzyme activity toward all tested AHLs, varying in length and nature of the substitution at the C3 position of the acyl chain. The data also indicate that the amide group and the ketone at the C1 position of the acyl chain of AHLs could be important structural features in enzyme-substrate interaction. Surprisingly, although carrying a (104)HX- HXDH(109) short sequence identical to the zinc-binding motif of several groups of metallohydrolytic enzymes, AHL-lactonase does not contain or require zinc or other metal ions for enzyme activity. Except for the amino acid residue His-104, which was shown previously to not be required for catalysis, kinetic study and conformational analysis using circular dichroism spectrometry showed that substitution of the other key residues in the motif (His-106, Asp-108, and His 109), as well as His-169 with serine, respectively, caused conformational changes and significant loss of enzyme activity. We conclude that AHL-lactonase is a highly specific enzyme and that the (106)HXDH(109) approximately H(169) of AHL lactonase represents a novel catalytic motif, which does not rely on zinc or other metal ions for activity. PMID- 14734560 TI - Translation initiation from a naturally occurring non-AUG codon in Saccharomyces cerevisiae. AB - Although previous studies have already shown that both cytoplasmic and mitochondrial activities of glycyl-tRNA synthetase are provided by a single gene, GRS1,in the yeast Saccharomyces cerevisiae, the mechanism by which this occurs remains unclear. Evidence presented here indicates that this bifunctional property is actually a result of two distinct translational products alternatively generated from a single transcript of this gene. Except for an amino-terminal 23-amino acid extension, these two isoforms have the same polypeptide sequence and function exclusively in their respective compartments under normal conditions. Reporter gene assays further suggest that this leader peptide can function independently as a mitochondrial targeting signal and plays the major role in the subcellular localization of the isoforms. Additionally, whereas the short protein is translationally initiated from a traditional AUG triplet, the longer isoform is generated from an upstream inframe UUG codon. To our knowledge, GRS1 appears to be the first example in the yeast wherein a functional protein isoform is initiated from a naturally occurring non-AUG codon. The results suggest that non-AUG initiation might be a mechanism existing throughout all kingdoms. PMID- 14734561 TI - Identification and biochemical characterization of a Werner's syndrome protein complex with Ku70/80 and poly(ADP-ribose) polymerase-1. AB - Werner's syndrome (WS) is an inherited disease characterized by genomic instability and premature aging. The WS gene encodes a protein (WRN) with helicase and exonuclease activities. We have previously reported that WRN interacts with Ku70/80 and this interaction strongly stimulates WRN exonuclease activity. To gain further insight on the function of WRN and its relationship with the Ku heterodimer, we established a cell line expressing tagged WRN(H), a WRN point mutant lacking helicase activity, and used affinity purification, immunoblot analysis and mass spectroscopy to identify WRN-associated proteins. To this end, we identified three proteins that are stably associated with WRN in nuclear extracts. Two of these proteins, Ku70 and Ku80, were identified by immunoblot analysis. The third polypeptide, which was identified by mass spectrometry analysis, is identical to poly(ADP-ribose) polymerase-1(PARP-1), a 113-kDa enzyme that functions as a sensor of DNA damage. Biochemical fractionation studies and immunoprecipitation assays and studies confirmed that endogenous WRN is associated with subpopulations of PARP-1 and Ku70/80 in the cell. Protein interaction assays with purified proteins further indicated that PARP-1 binds directly to WRN and assembles in a complex with WRN and Ku70/80. In the presence of DNA and NAD(+), PARP-1 poly(ADP-ribosyl)ates itself and Ku70/80 but not WRN, and gel-shift assays showed that poly-(ADP-ribosyl)ation of Ku70/80 decreases the DNA-binding affinity of this factor. Significantly, (ADP ribosyl)ation of Ku70/80 reduces the ability of this factor to stimulate WRN exonuclease, suggesting that covalent modification of Ku70/80 by PARP-1 may play a role in the regulation of the exonucleolytic activity of WRN. PMID- 14734562 TI - Activation and interaction of ATF2 with the coactivator ASC-2 are responsive for granulocytic differentiation by retinoic acid. AB - Terminal differentiation of hematopoietic cells follows a precisely orchestrated program of transcriptional regulatory events at the promoters of both lineage specific and ubiquitous genes. Here we show that the transcription factor ATF2 is associated with the induction of granulocytic differentiation, and the molecular interaction of ATF2 with a tissue-specific coactivator activating signal cointegator-2 (ASC-2) potentiates the differentiation procedure. All-trans retinoic acid (RA) induced the phosphorylation and expression of ATF2 in the early and middle phase of granulocyte differentiation, respectively. The activation of granulocyte-specific gene expression is increased with the concerted action of another basic regionleucine zipper factor, CCAAT/enhancer binding protein (C/EBPalpha), and ASC-2, which function in a cooperative manner. The interaction between ATF2 and C/EBPalpha in RA-treated cells was enhanced by the ectopic expression of ASC-2. ATF2-mediated transactivation was also increased by co-transfection of ASC-2. This resulted from the direct protein interaction that the N-terminal transactivation domain of ATF2 interacts with the central region of ASC-2. Furthermore, the molecular interaction of ATF2 and ASC-2 was stimulated by RA treatment and inhibited by p38beta kinase inhibitor. Taking these results together, these results suggest that the differentiation-dependent expression and phosphorylation of ATF2 protein physically and functionally interacts with C/EBPalpha and coativator ASC-2 and synergizes to induce target gene transcription during granulocytic differentiation. PMID- 14734563 TI - Stopped-flow fluorescence analysis of the conformational changes in the GroEL apical domain: relationships between movements in the apical domain and the quaternary structure of GroEL. AB - GroEL undergoes numerous conformational alterations in the course of facilitating the folding of various proteins, and the specific movements of the GroEL apical domain are of particular importance in the molecular mechanism. In order to monitor in detail the numerous movements of the GroEL apical domain, we have constructed a mutant chaperonin (GroEL R231W) with wild type-like function and a fluorescent probe introduced into the apical domain. By monitoring the tryptophan fluorescence changes of GroEL R231W upon ATP addition in the presence and absence of the co-chaperonin GroES, we detected a total of four distinct kinetic phases that corresponded to conformational changes of the apical domain and GroES binding. By introducing this mutation into a single ring variant of GroEL (GroEL SR-1), we determined the extent of inter-ring cooperation that was involved in apical domain movements. Surprisingly, we found that the apical domain movements of GroEL were affected only slightly by the change in quaternary structure. Our experiments provide a number of novel insights regarding the dynamic movements of this protein. PMID- 14734565 TI - Two distinct crt gene clusters for two different functional classes of carotenoid in Bradyrhizobium. AB - Aerobic photosynthetic bacteria possess the unusual characteristic of producing different classes of carotenoids. In this study, we demonstrate the presence of two distinct crt gene clusters involved in the synthesis of spirilloxanthin and canthaxanthin in a Bradyrhizobium strain. Each cluster contains the genes crtE, crtB, and crtI leading to the common precursor lycopene. We show that spirilloxanthin is associated with the photosynthetic complexes, while canthaxanthin protects the bacteria from oxidative stress. Only the spirilloxanthin crt genes are regulated by light via the control of a bacteriophytochrome. Despite this difference in regulation, the biosyntheses of both carotenoids are strongly interconnected at the level of the common precursors. Phylogenetic analysis suggests that the canthaxanthin crt gene cluster has been acquired by a lateral gene transfer. This acquisition may constitute a major selective advantage for this class of bacteria, which photosynthesize only under conditions where harmful reactive oxygen species are generated. PMID- 14734564 TI - Dissecting transcription-coupled and global genomic repair in the chromatin of yeast GAL1-10 genes. AB - Transcription-coupled repair (TCR) and global genomic repair (GGR) of UV-induced cyclobutane pyrimidine dimers were investigated in the yeast GAL1-10 genes. Both Rpb9- and Rad26-mediated TCR are confined to the transcribed strands, initiating at upstream sites approximately 100 nucleotides from the upstream activating sequence shared by the two genes. However, TCR initiation sites do not correlate with either transcription start sites or TATA boxes. Rad16-mediated GGR tightly correlates with nucleosome positioning when the genes are repressed and are slow in the nucleosome core and fast in linker DNA. Induction of transcription enhanced GGR in nucleosome core DNA, especially in the nucleosomes around and upstream of the transcription start sites. Furthermore, when the genes were induced, GGR was slower in the transcribed regions than in the upstream regions. Finally, simultaneous deletion of RAD16, RAD26, and RPB9 resulted in no detectable repair in all sites along the region analyzed. Our results suggest that (a). TCR may be initiated by a transcription activator, presumably through the loading of RNA polymerase II, rather than by transcription initiation or elongation per se; (b). TCR and nucleosome disruption-enhanced GGR are the major causes of rapid repair in regions around and upstream of transcription start sites; (c). transcription machinery may hinder access of NER factors to a DNA lesion in the absence of a transcription-repair coupling factor; and (d). other than GGR mediated by Rad16 and TCR mediated by Rad26 and Rpb9, no other nucleotide excision repair pathway exists in these RNA polymerase II-transcribed genes. PMID- 14734566 TI - Suramin derivatives as inhibitors and activators of protein-tyrosine phosphatases. AB - Protein-tyrosine phosphatases (PTPs) are important signaling enzymes that have emerged within the last decade as a new class of drug targets. It has previously been shown that suramin is a potent, reversible, and competitive inhibitor of PTP1B and Yersinia PTP (YopH). We therefore screened 45 suramin analogs against a panel of seven PTPs, including PTP1B, YopH, CD45, Cdc25A, VHR, PTPalpha, and LAR, to identify compounds with improved potency and specificity. Of the 45 compounds, we found 11 to have inhibitory potency comparable or significantly improved relative to suramin. We also found suramin to be a potent inhibitor (IC(50) = 1.5 microm) of Cdc25A, a phosphatase that mediates cell cycle progression and a potential target for cancer therapy. In addition we also found three other compounds, NF201, NF336, and NF339, to be potent (IC(50) < 5 microm) and specific (at least 20-30-fold specificity with respect to the other human PTPs tested) inhibitors of Cdc25A. Significantly, we found two potent and specific inhibitors, NF250 and NF290, for YopH, the phosphatase that is an essential virulence factor for bubonic plague. Two of the compounds tested, NF504 and NF506, had significantly improved potency as PTP inhibitors for all phosphatases tested except for LAR and PTPalpha. Surprisingly, we found that a significant number of these compounds activated the receptor-like phosphatases, PTPalpha and LAR. In further characterizing this activation phenomenon, we reveal a novel role for the membrane-distal cytoplasmic PTP domain (D2) of PTPalpha: the direct intramolecular regulation of the activity of the membrane-proximal cytoplasmic PTP domain (D1). Binding of certain of these compounds to PTPalpha disrupts D1-D2 basal state contacts and allows new contacts to occur between D1 and D2, which activates D1 by as much as 12-14-fold when these contacts are optimized. PMID- 14734567 TI - Threonine 98, the pivotal residue of tissue inhibitor of metalloproteinases (TIMP)-1 in metalloproteinase recognition. AB - Tissue inhibitors of metalloproteinases (TIMPs) are the endogenous modulators of the zinc-dependent mammalian matrix metalloproteinases (MMPs) and their close associates, proteinases of the ADAM (a disintegrin and metalloproteinase) and ADAM with thrombospondin repeats families. There are four variants of TIMPs, and each has its defined set of metalloproteinase (MP) targets. TIMP-1, in particular, is inactive against several of the membrane-type MMPs (MT-MMPs), MMP 19, and the ADAM proteinase TACE (tumor necrosis factor-alpha-converting enzyme, ADAM-17). The molecular basis for such inactivity is unknown. Previously, we showed that TIMP-1 could be transformed into an active inhibitor against MT1-MMP by the replacement of threonine 98 residue with leucine (T98L). Here, we reveal that the T98L mutation has in fact transformed TIMP-1 into a versatile inhibitor against an array of MPs otherwise insensitive to wild-type TIMP-1; examples include TACE, MMP-19, and MT5-MMP. Using T98L as the scaffold, we created a TIMP 1 variant that is fully active against TACE. The binding affinity of the mutant (V4S/TIMP-3-AB-loop/V69L/T98L) (K (app)(i) 0.14 nm) surpassed that of TIMP-3 (K (app)(i) 0.22 nm), the only natural TIMP inhibitor of the enzyme. The requirement for leucine is absolute for the transformation in inhibitory pattern. On the other hand, the mutation has minimal impact on the MPs already well inhibited by wild-type TIMP-1, such as gelatinase-A and stromelysin-1. Not only have we unlocked the molecular basis for the inactivity of TIMP-1 against several of the MPs, but also our findings fundamentally modify the current beliefs on the molecular mechanism of TIMP-MP recognition and selectivity. PMID- 14734568 TI - MeaB is a component of the methylmalonyl-CoA mutase complex required for protection of the enzyme from inactivation. AB - Adenosylcobalamin-dependent methylmalonyl-CoA mutase catalyzes the interconversion of methylmalonyl-CoA and succinyl-CoA. In humans, deficiencies in the mutase lead to methylmalonic aciduria, a rare disease that is fatal in the first year of life. Such inherited deficiencies can result from mutations in the mutase structural gene or from mutations that impair the acquisition of cobalamins. Recently, a human gene of unknown function, MMAA, has been implicated in methylmalonic aciduria (Dobson, C. M., Wai, T., Leclerc, D., Wilson, A., Wu, X., Dore, C., Hudson, T., Rosenblatt, D. S., and Gravel, R. A. (2002) Proc. Natl. Acad. Sci. U. S. A. 99, 15554-15559). MMAA orthologs are widespread in bacteria, archaea, and eukaryotes. In Methylobacterium extorquens AM1, a mutant defective in the MMAA homolog meaB was unable to grow on C(1) and C(2) compounds because of the inability to convert methylmalonyl-CoA to succinyl-CoA (Korotkova N., Chistoserdova, L., Kuksa, V., and Lidstrom, M. E. (2002) J. Bacteriol. 184, 1750 1758). Here we demonstrate that this defect is not due to the absence of adenosylcobalamin but due to an inactive form of methylmalonyl-CoA mutase. The presence of active mutase in double mutants defective in MeaB and in the synthesis of either R-methylmalonyl-CoA or adenosylcobalamin indicates that MeaB is necessary for protection of mutase from inactivation during catalysis. MeaB and methylmalonyl-CoA mutase from M. extorquens were cloned and purified in their active forms. We demonstrated that MeaB forms a complex with methylmalonyl-CoA mutase and stimulates in vitro mutase activity. These results support the hypothesis that MeaB functions to protect methylmalonyl-CoA mutase from irreversible inactivation. PMID- 14734569 TI - Mutations in the yeast mrf1 gene encoding mitochondrial release factor inhibit translation on mitochondrial ribosomes. AB - Although the control of mitochondrial translation in the yeast Saccharomyces cerevisiae has been studied extensively, the mechanism of termination remains obscure. Ten mutations isolated in a genetic screen for read-through of premature stop codons in mitochondrial genes were localized in the chromosomal gene encoding the mitochondrial release factor mRF1. The mrf1-13 and mrf1-780 mutant genes, in contrast to other alleles, caused a non-respiratory phenotype that correlated with decreased expression of mitochondrial genes as well as a reporter ARG8(m) gene inserted into mitochondrial DNA. The steady-state levels of several mitochondrially encoded proteins, but not their mRNAs, were dramatically decreased in mrf1-13 and mrf1-780 cells. Structural models of mRF1 were constructed, allowing localization of residues substituted in the mrf1 mutants and offering an insight into the possible mechanism by which these mutations change the mitochondrial translation termination fidelity. Inhibition of mitochondrial translation in mrf1-13 and mrf1-780 correlated with the three dimensional localization of the mutated residues close to the PST motif presumably involved in the recognition of stop codons in mitochondrial mRNA. PMID- 14734571 TI - A piece of my mind. Stitches. PMID- 14734570 TI - Annexin 2 binding to phosphatidylinositol 4,5-bisphosphate on endocytic vesicles is regulated by the stress response pathway. AB - Annexin 2 is a Ca(2+)-binding protein that has an essential role in actin dependent macropinosome motility. We show here that macropinosome rocketing can be induced by hyperosmotic shock, either alone or synergistically when combined with phorbol ester or pervanadate. Rocketing was blocked by inhibitors of phosphatidylinositol-3-kinase(s), p38 mitogen-activated protein (MAP) kinase, and calcium, suggesting the involvement of phosphoinositide signaling. Since various phosphoinositides are enriched on inwardly mobile vesicles, we examined whether or not annexin 2 binds to any of this class of phospholipid. In liposome sedimentation assays, we show that recombinant annexin 2 binds to phosphatidylinositol 4,5-bisphosphate (PtdIns-4,5P(2)) but not to other poly- and mono-phosphoinositides. The affinity of annexin 2 for PtdIns-4,5P(2) (K(D) approximately 5 microm) is comparable with those reported for a variety of PtdIns 4,5P(2)-binding proteins and is enhanced in the presence of Ca(2+). Although annexin 1 also bound to PtdIns-4,5P(2), annexin 5 did not, indicating that this is not a generic annexin property. To test whether annexin 2 binds to PtdIns 4,5P(2) in vivo, we microinjected rat basophilic leukemia cells stably expressing annexin 2-green fluorescent protein (GFP) with fluorescently tagged antibodies to PtdIns-4,5P(2). Annexin 2-GFP and anti-PtdIns-4,5P(2) IgG co-localize at sites of pinosome formation, and annexin 2-GFP relocalizes to intracellular membranes in Ptk cells microinjected with Arf6Q67L, which has been shown to stimulate PtdIns 4,5P(2) synthesis on pinosomes through activation of phosphatidylinositol 5 kinase. These results establish a novel phospholipid-binding specificity for annexin 2 consistent with a role in mediating the interaction between the macropinosome surface and the polymerized actin tail. PMID- 14734572 TI - Physicians apply genome research to treating critical illness and injury. PMID- 14734573 TI - Fear factor: scientists probe how the mind overcomes fear itself. PMID- 14734581 TI - Traditional risk factors for coronary heart disease. PMID- 14734582 TI - Traditional risk factors for coronary heart disease. PMID- 14734583 TI - Traditional risk factors for coronary heart disease. PMID- 14734584 TI - Novel risk factors for atherosclerosis. PMID- 14734585 TI - Novel risk factors for atherosclerosis. PMID- 14734586 TI - Antibiotics and coronary heart disease. PMID- 14734587 TI - Antibiotics and coronary heart disease. PMID- 14734588 TI - Serologic factors in autoimmune ear disease. PMID- 14734589 TI - Definitions of medical injuries. PMID- 14734590 TI - Definitions of medical injuries. PMID- 14734591 TI - Definitions of medical injuries. PMID- 14734592 TI - Hemangiomas and other congenital malformations in infants exposed to antiretroviral therapy in utero. PMID- 14734593 TI - Effects of ranolazine with atenolol, amlodipine, or diltiazem on exercise tolerance and angina frequency in patients with severe chronic angina: a randomized controlled trial. AB - CONTEXT: Many patients with chronic angina experience anginal episodes despite revascularization and antianginal medications. In a previous trial, antianginal monotherapy with ranolazine, a drug believed to partially inhibit fatty acid oxidation, increased treadmill exercise performance; however, its long-term efficacy and safety have not been studied in combination with beta-blockers or calcium antagonists in a large patient population with severe chronic angina. OBJECTIVES: To determine whether, at trough levels, ranolazine improves the total exercise time of patients who have symptoms of chronic angina and who experience angina and ischemia at low workloads despite taking standard doses of atenolol, amlodipine, or diltiazem and to determine times to angina onset and to electrocardiographic evidence of myocardial ischemia, effect on angina attacks and nitroglycerin use, and effect on long-term survival in an open-label observational study extension. DESIGN, SETTING, AND PATIENTS: A randomized, 3 group parallel, double-blind, placebo-controlled trial of 823 eligible adults with symptomatic chronic angina who were randomly assigned to receive placebo or 1 of 2 doses of ranolazine. Patients treated at the 118 participating ambulatory outpatient settings in several countries were enrolled in the Combination Assessment of Ranolazine In Stable Angina (CARISA) trial from July 1999 to August 2001 and followed up through October 31, 2002. INTERVENTION: Patients received twice-daily placebo or 750 mg or 1000 mg of ranolazine. Treadmill exercise 12 hours (trough) and 4 hours (peak) after dosing was assessed after 2, 6 (trough only), and 12 weeks of treatment. MAIN OUTCOME MEASURES: Change in exercise duration, time to onset of angina, time to onset of ischemia, nitroglycerin use, and number of angina attacks. RESULTS: Trough exercise duration increased by 115.6 seconds from baseline in both ranolazine groups (pooled) vs 91.7 seconds in the placebo group (P =.01). The times to angina and to electrocardiographic ischemia also increased in the ranolazine groups, at peak more than at trough. The increases did not depend on changes in blood pressure, heart rate, or background antianginal therapy and persisted throughout 12 weeks. Ranolazine reduced angina attacks and nitroglycerin use by about 1 per week vs placebo (P<.02). Survival of 750 patients taking ranolazine during the CARISA trial or its associated long-term open-label study was 98.4% in the first year and 95.9% in the second year. CONCLUSION: Twice-daily doses of ranolazine increased exercise capacity and provided additional antianginal relief to symptomatic patients with severe chronic angina taking standard doses of atenolol, amlodipine, or diltiazem, without evident adverse, long-term survival consequences over 1 to 2 years of therapy. PMID- 14734594 TI - Memantine treatment in patients with moderate to severe Alzheimer disease already receiving donepezil: a randomized controlled trial. AB - CONTEXT: Memantine is a low- to moderate-affinity, uncompetitive N-methyl-D aspartate receptor antagonist. Controlled trials have demonstrated the safety and efficacy of memantine monotherapy for patients with moderate to severe Alzheimer disease (AD) but no controlled trials of memantine in patients receiving a cholinesterase inhibitor have been performed. OBJECTIVE: To compare the efficacy and safety of memantine vs placebo in patients with moderate to severe AD already receiving stable treatment with donepezil. DESIGN, SETTING, AND PARTICIPANTS: A randomized, double-blind, placebo-controlled clinical trial of 404 patients with moderate to severe AD and Mini-Mental State Examination scores of 5 to 14, who received stable doses of donepezil, conducted at 37 US sites between June 11, 2001, and June 3, 2002. A total of 322 patients (80%) completed the trial. INTERVENTIONS: Participants were randomized to receive memantine (starting dose 5 mg/d, increased to 20 mg/d, n = 203) or placebo (n = 201) for 24 weeks. MAIN OUTCOME MEASURES: Change from baseline on the Severe Impairment Battery (SIB), a measure of cognition, and on a modified 19-item AD Cooperative Study-Activities of Daily Living Inventory (ADCS-ADL19). Secondary outcomes included a Clinician's Interview-Based Impression of Change Plus Caregiver Input (CIBIC-Plus), the Neuropsychiatric Inventory, and the Behavioral Rating Scale for Geriatric Patients (BGP Care Dependency Subscale). RESULTS: The change in total mean (SE) scores favored memantine vs placebo treatment for SIB (possible score range, 0 100), 0.9 (0.67) vs -2.5 (0.69), respectively (P<.001); ADCS-ADL19 (possible score range, 0-54), -2.0 (0.50) vs -3.4 (0.51), respectively (P =.03); and the CIBIC-Plus (possible score range, 1-7), 4.41 (0.074) vs 4.66 (0.075), respectively (P =.03). All other secondary measures showed significant benefits of memantine treatment. Treatment discontinuations because of adverse events for memantine vs placebo were 15 (7.4%) vs 25 (12.4%), respectively. CONCLUSIONS: In patients with moderate to severe AD receiving stable doses of donepezil, memantine resulted in significantly better outcomes than placebo on measures of cognition, activities of daily living, global outcome, and behavior and was well tolerated. These results, together with previous studies, suggest that memantine represents a new approach for the treatment of patients with moderate to severe AD. PMID- 14734595 TI - Surveillance of medical device-related hazards and adverse events in hospitalized patients. AB - CONTEXT: Although adverse drug events have been extensively evaluated by computer based surveillance, medical device errors have no comparable surveillance techniques. OBJECTIVES: To determine whether computer-based surveillance can reliably identify medical device-related hazards (no known harm to patient) and adverse medical device events (AMDEs; patient experienced harm) and to compare alternative methods of detection of device-related problems. DESIGN, SETTING, AND PARTICIPANTS: This descriptive study was conducted from January through September 2000 at a 520-bed tertiary teaching institution in the United States with experience in using computer tools to detect and prevent adverse drug events. All 20 441 regular and short-stay patients (excluding obstetric and newborn patients) were included. MAIN OUTCOME MEASURES: Medical device events as detected by computer-based flags, telemetry problem checklists, International Classification of Diseases, Ninth Revision (ICD-9) discharge code (which could include AMDEs present at admission), clinical engineering work logs, and patient survey results were compared with each other and with routine voluntary incident reports to determine frequencies, proportions, positive predictive values, and incidence rates by each technique. RESULTS: Of the 7059 flags triggered, 552 (7.8%) indicate a device-related hazard or AMDE. The estimated 9-month incidence rates (number per 1000 admissions [95% confidence intervals]) for AMDEs were 1.6 (0.9 2.5) for incident reports, 27.7 (24.9-30.7) for computer flags, and 64.6 (60.4 69.1) for ICD-9 discharge codes. Few of these events were detected by more than 1 surveillance method, giving an overall incidence of AMDE detected by at least 1 of these methods of 83.7 per 1000 (95% confidence interval, 78.8-88.6) admissions. The positive predictive value of computer flags for detecting device related hazards and AMDEs ranged from 0% to 38%. CONCLUSIONS: More intensive surveillance methods yielded higher rates of medical device problems than found with traditional voluntary reporting, with little overlap between methods. Several detection methods had low efficiency in detecting AMDEs. The high rate of AMDEs suggests that AMDEs are an important patient safety issue, but additional research is necessary to identify optimal AMDE detection strategies. PMID- 14734596 TI - Poor control of risk factors for vascular disease among adults with previously diagnosed diabetes. AB - CONTEXT: Control of blood glucose levels, blood pressure, and cholesterol levels is proven to reduce the risk of vascular disease among individuals with diabetes mellitus; however, the current state of control of these risk factors among individuals in the United States is uncertain. OBJECTIVES: To examine 1999-2000 national data on control of risk factors for vascular disease among adults with previously diagnosed diabetes and to assess trends during the past decade. DESIGN, SETTING, AND PARTICIPANTS: Review of data from the Third National Health and Nutrition Examination Survey (NHANES III, conducted 1988-1994) and NHANES 1999-2000, cross-sectional surveys of a nationally representative sample of the noninstitutionalized civilian US population. Participants were adults aged 20 years and older with previously diagnosed diabetes who participated in both the interview and examination in either NHANES III (n = 1265) or NHANES 1999-2000 (n = 441). MAIN OUTCOME MEASURES: Levels of glycosylated hemoglobin (HbA1c), blood pressure, and total serum cholesterol in reference to target goals. RESULTS: Compared with NHANES III, participants with previously diagnosed diabetes in NHANES 1999-2000 were similar by age and sex, were less likely to be non-Hispanic white, were diagnosed at an earlier age, had a higher body mass index, and were more likely to use insulin in combination with oral agents. In NHANES 1999-2000, only 37.0% of participants achieved the target goal of HbA1c level less than 7.0% and 37.2% of participants were above the recommended "take action" HbA1c level of greater than 8.0%; these percentages did not change significantly from NHANES III (P =.11 and P =.87, respectively). Only 35.8% of participants achieved the target of systolic blood pressure (SBP) less than 130 mm Hg and diastolic blood pressure (DBP) less than 80 mm Hg, and 40.4% had hypertensive blood pressure levels (SBP > or =140 or DBP > or =90 mm Hg). These percentages did not change significantly from NHANES III (P =.10 and P =.56, respectively). Over half (51.8%) of the participants in NHANES 1999-2000 had total cholesterol levels of 200 mg/dL or greater (vs 66.1% in NHANES III; P<.001). In total, only 7.3% (95% confidence interval, 2.8%-11.9%) of adults with diabetes in NHANES 1999-2000 attained recommended goals of HbA1c level less than 7%, blood pressure less than 130/80 mm Hg, and total cholesterol level less than 200 mg/dL (5.18 mmol/L). CONCLUSION: Further public health efforts are needed to control risk factors for vascular disease among individuals with diagnosed diabetes. PMID- 14734597 TI - Car occupant death according to the restraint use of other occupants: a matched cohort study. AB - CONTEXT: A car occupant could be killed if struck by another occupant who was catapulted forward, backward, or sideways in a crash. OBJECTIVE: To estimate the association between death of a car occupant (the target) and restraint use by other occupants. DESIGN: Matched-pair cohort study comparing the outcomes of 2 target occupants in the same passenger car that crashed. SETTING: United States traffic crashes in 1988-2000, using data from the Fatality Analysis Reporting System. SUBJECTS: Target pairs, at least 1 of whom died: 61 834 front-seat pairs, 5278 rear-seat pairs, and 21 127 pairs on the left or right side. MAIN OUTCOME MEASURES: Adjusted risk ratio (RR) for death within 30 days of a crash. RESULTS: The risk of death was greater for a restrained front target occupant in front of an unrestrained occupant compared with a restrained front target in front of a restrained occupant (adjusted RR, 1.20; 95% confidence interval [CI], 1.10-1.31). For a restrained rear target occupant behind an unrestrained occupant compared with a restrained rear target occupant behind a restrained occupant, the adjusted RR was 1.22 (95% CI, 1.10-1.36). For a restrained side target occupant sitting next to an unrestrained occupant compared with a restrained side target occupant sitting next to a restrained occupant, the adjusted RR was 1.15 (95% CI, 1.08 1.22). Among unrestrained target occupants, the adjusted RRs were, for front targets, 1.04 (95% CI, 0.97-1.12), rear targets, 1.22 (95% CI, 1.10-1.36), and side targets, 0.85 (95% CI, 0.80-0.92). CONCLUSION: Persons who wish to reduce their risk of death in a crash should wear their own restraint and should ask others in the same car to use their restraints. PMID- 14734598 TI - Arterial puncture closing devices compared with standard manual compression after cardiac catheterization: systematic review and meta-analysis. AB - CONTEXT: Arterial puncture closing devices (APCDs) were developed to replace standard compression at the puncture site and to shorten bed rest following percutaneous coronary intervention. OBJECTIVE: To assess the safety and efficacy of APCDs (Angioseal, Vasoseal, Duett, Perclose, Techstar, Prostar) compared with standard manual compression in patients undergoing coronary angiography or percutaneous vascular interventions. DATA SOURCES: A systematic literature search of MEDLINE (1966-January 2003), EMBASE (1989-January 2003), PASCAL (1996-January 2003), BIOSIS (1990-January 2003), and CINHAL (1982-January 2003) databases and the Cochrane Central Register of Controlled Trials for relevant articles in any language. STUDY SELECTION: Included randomized controlled trials reporting vascular complications at the puncture site (hematoma, bleeding, arteriovenous fistula, pseudoaneurysm) and efficacy (time to hemostasis, time to ambulation, time to discharge from hospital). DATA EXTRACTION: Two reviewers abstracted the data independently and in duplicate. Disagreements were resolved by discussion among at least 3 reviewers. The most important criteria were adequacy of allocation concealment, whether the analysis was according to the intention-to treat principle, and if person assessing the outcome was blinded to intervention group. Random-effects models were used to pool the data. DATA SYNTHESIS: Thirty trials met the selection criteria and included up to 4000 patients. When comparing any APCD with standard compression, the relative risk (RR) of groin hematoma was 1.14 (95% confidence interval [CI], 0.86-1.51; P =.35); bleeding, 1.48 (95% CI, 0.88-2.48; P =.14); developing an arteriovenous fistula, 0.83 (95% CI, 0.23-2.94; P =.77); and developing a pseudoaneurysm at the puncture site, 1.19 (95% CI, 0.75-1.88; P =.46). Time to hemostasis was shorter in the group with APCD compared with standard compression (mean difference, 17 minutes; range, 14-19 minutes), but there was a high degree of heterogeneity among studies. Only 2 studies explicitly reported allocation concealment, blinded outcome assessment, and intention-to-treat analysis. When limiting analyses to only trials that used explicit intention-to-treat approaches, APCDs were associated with a higher risk of hematoma (RR, 1.89; 95% CI, 1.13-3.15) and a higher risk of pseudoaneurysm (RR, 5.40; 95% CI, 1.21-24.5). CONCLUSIONS: Based on this meta-analysis of 30 randomized trials, many of poor methodological quality, there is only marginal evidence that APCDs are effective and there is reason for concern that these devices may increase the risk of hematoma and pseudoaneurysm. PMID- 14734599 TI - Neuroprotection in Parkinson disease: mysteries, myths, and misconceptions. AB - Parkinson disease is an age-related neurodegenerative disease that affects approximately 1 million persons in the United States. Current therapies provide effective control of symptoms, particularly in the early stages of the disease, but most patients develop motor complications with long-term treatment, and features develop such as postural instability, falling, and dementia that are not adequately controlled with existing medications. Accordingly, neuroprotective therapy that might slow, stop, or reverse disease progression is urgently needed. While many agents appear to be promising based on laboratory studies, selecting clinical end points for clinical trials that are not confounded by symptomatic effects of the study intervention has been difficult. More recently, neuroimaging end points have been used as biomarkers of disease progression, but again there are concerns that they may be influenced by regulatory effects of the drugs used. We review clinical trials aimed at detecting neuroprotection in Parkinson disease and address the controversies surrounding the interpretation of these studies. PMID- 14734600 TI - Ranolazine and other antianginal therapies in the era of the drug-eluting stent. PMID- 14734601 TI - Medical device-associated safety and risk: surveillance and stratagems. PMID- 14734602 TI - Access to JAMA. PMID- 14734603 TI - JAMA patient page. Parkinson disease. PMID- 14734604 TI - A species-specific determinant on beta2-microglobulin required for Ly49A recognition of its MHC class I ligand. AB - The mouse inhibitory NK cell receptor Ly49A recognizes the mouse MHC class I molecule H-2D(k). The present study focuses on the species specificity of beta(2) microglobulin (beta(2)m), an invariant component of MHC class I, in the interaction between Ly49A and H-2D(k). Transfection of the beta(2)m-defective mouse cell line R1E/TL8x.1 with human (h) beta(2)m induced cell-surface expression of H-2D(k), but failed to protect the cells from killing by Ly49A(+) NK cells. In contrast, the cells transfected with mouse (m) beta(2)m were protected from killing by Ly49A(+) NK cells. These data indicate that Ly49A distinguishes mbeta(2)m from hbeta(2)m when it recognizes the H-2D(k) complexes. To identify the species-specific determinant of beta(2)m required for Ly49A recognition of H-2D(k), we prepared a panel of mbeta(2)m mutants and tested the H 2D(k) that included each of the beta(2)m mutants for its capacity to engage Ly49A on NK cells. Ly49A failed to functionally recognize the H-2D(k) that included the mbeta(2)m with K3R and Q29G mutations. Moreover, Ly49A was able to recognize the H-2D(k) that included the hbeta(2)m with R3K and G29Q mutations. These data indicate that Lys3 and Gln29 consist of the central part of the species-specific determinant of beta(2)m required for Ly49A recognition of H-2D(k). The two residues are conserved in the mouse and the rat, in which NK cells use Ly49 family molecules as the receptors specific for MHC class I. These results suggest functional importance of beta(2)m in NK cell recognition of target cells. PMID- 14734605 TI - Blocking inducible co-stimulator in the absence of CD28 impairs Th1 and CD25+ regulatory T cells in murine colitis. AB - Several autoimmune disease models depend on an imbalance in the activation of aggressor T(h)1 and CD4(+)CD25(+) regulatory T (T(reg)) cells. Here we compare the requirement for signals through the co-stimulatory molecules CD28 and inducible co-stimulator (ICOS) in chronic murine colitis, a model for inflammatory bowel disease. We used a colitis model in which disease-causing CD45RB(hi) T cells alone or in combination with CD4(+)CD25(+) T cells from either CD28-deficient or wild-type donors were transferred into T cell-deficient animals, half of which were treated with ICOS-blocking reagents. Blocking ICOS on the surface of CD28-deficient T(h)1 cells abrogated development of colitis, whereas blocking CD28 or ICOS alone had little to no effect on disease induction. In contrast to T(h)1 cells, regulatory T cell functioning depended mostly on CD28 signaling with only a minor contribution for ICOS. We conclude that CD28 and ICOS collaborate to development of murine colitis by aggressor T(h)1 cells, and that CD28 is required for T(reg) cells, which should caution against the use of CD28 blocking reagents in inflammatory bowel disease. PMID- 14734606 TI - Impaired IFN-gamma production of Valpha24 NKT cells in non-remitting sarcoidosis. AB - Sarcoidosis is a systemic disorder associated with granuloma characterized by an abnormal T(h)1-type cytokine production and accumulation of T(h)1 CD4 T cells in the granuloma lesions, suggesting an importance of T(h)1 responses in sarcoidosis. However, the pathogenesis of sarcoidosis remains to be solved. Here, we investigated the nature of V(alpha)24 NKT cells with immunoregulatory functions in sarcoidosis. Patients with non-remitting sarcoidosis displayed a decrease in the number of V(alpha)24 NKT cells in peripheral blood, but an accumulation of these cells in granulomatous lesions. When stimulated with the specific glycolipid ligand, alpha-galactosylceramide, peripheral blood V(alpha)24 NKT cells from patients with non-remitting disease produced significantly less IFN-gamma than those from healthy volunteers, but normal levels of IL-4. The reduced IFN-gamma production was observed only in V(alpha)24 NKT cells and not conventional CD4 T cells, but was normal in patients with remitting disease, suggesting that non-remitting sarcoidosis involves an insufficient IFN-gamma production of V(alpha)24 NKT cells which is well correlated with disease activity. Thus, these results suggest that V(alpha)24 NKT cells play a crucial role in the disease status of sarcoidosis. PMID- 14734607 TI - Accumulation of Valpha7.2-Jalpha33 invariant T cells in human autoimmune inflammatory lesions in the nervous system. AB - T cells expressing an invariant TCR alpha chain and NK cell markers are expected to exhibit unique functions. Whereas much attention has been paid to CD1d restricted NKT cells, a second NKT cell population bearing an invariant AV19-AJ33 TCR has recently been identified in mice and humans. Selection and/or expansion of this population require B cells, and would involve a non-classical class I related molecule MR1. Although their preferential distribution in the gut mucosa indicates their role in the host response at the site of pathogen entry, it remains unknown whether they play an alternative role at different sites or in immunological disorders. Using single-strand conformation polymorphism clonotype analysis, we investigated the presence of the human AV19-AJ33 T cells (V(alpha)7.2-J(alpha)33 T cells) in autopsy samples from multiple sclerosis (MS) patients as well as in nerve biopsy samples from chronic inflammatory demyelinating polyneuropathy (CIDP) patients. Here we report that the V(alpha)7.2 J(alpha)33 T cells are accumulated in some of the central nervous system lesions of MS and in the majority of the peripheral nerve samples from CIDP. We have previously revealed that CD1d-restricted, V(alpha)24-J(alpha)Q NKT cells are remarkably reduced in the peripheral blood from MS. However, V(alpha)7.2 J(alpha)33 T cells are not reduced in the peripheral blood from MS and could be detected in a large majority of the cerebrospinal fluid samples obtained during relapse of MS. The present results indicate that the V(alpha)7.2-J(alpha)33 T cells are involved in the autoimmune inflammatory lesions. PMID- 14734608 TI - Coronin-1 expression in T lymphocytes: insights into protein function during T cell development and activation. AB - Coronin has been described as an actin-binding protein of Dictyostelium discoideum, and it has been demonstrated to play a role in cell migration, cytokinesis and phagocytosis. Coronin-related proteins are found in many eukaryotic species, including Coronin-1 in mammals whose expression is enriched in the hematopoietic tissues. Here, we characterize Coronin-1 gene and protein expression in mouse embryonic and adult T lymphocytes. Coronin-1 is expressed throughout T cell ontogeny and in peripheral alphabeta T cells. Expression varies along thymic cell development, with maximum levels observed in embryonic early thymocytes and, in the adults, the selected TCRalphabeta(+) single-positive thymocytes. Subcellular localization analysis indicates that Coronin-1 is in equilibrium between the cytosol and the cell cortex, where it accumulates in F actin-rich membrane protrusions induced by polarized activation of TCR-CD3 stimulated T cells. These data are consistent with a role of Coronin-1 in T cell differentiation/activation events involving membrane dynamisms and the cortical actin cytoskeleton. PMID- 14734609 TI - Down-regulation of the invariant Valpha14 antigen receptor in NKT cells upon activation. AB - NKT cells expressing the invariant Valpha14 antigen receptor constitute a novel lymphocyte subpopulation with immunoregulatory functions. Stimulation via their invariant Valpha14 receptor with anti-CD3 or a ligand, alpha-galactosylceramide (alpha-GalCer), triggers activation of Valpha14 NKT cells, resulting in a rapid cytokine production such as IFN-gamma and IL-4. Soon after their receptor activation, Valpha14 NKT cells disappeared as judged by staining with CD1d tetramer loaded with alpha-GalCer (alpha-GalCer/CD1d tetramer), which has been believed to be due to apoptotic cell death. Here we show that such a disappearance was largely attributed to down-regulation of the Valpha14 receptor. In fact, Valpha14 NKT cells were relatively resistant to apoptosis compared to the conventional T cells as evidenced by less staining with Annexin-V, a limited DNA fragmentation, and their preferential expression of anti-apoptotic genes such as NAIP and MyD118. Furthermore, they did not become tolerant, and maintained their proliferative capacity and cytokine production even after their receptor down-regulation. These as yet unrecognized facets of Valpha14 NKT cells are discussed in relation to their regulatory functions. PMID- 14734610 TI - IL-10 is involved in the suppression of experimental autoimmune encephalomyelitis by CD25+CD4+ regulatory T cells. AB - CD25(+)CD4(+) regulatory T cells inhibit the activation of autoreactive T cells in vitro and in vivo, and suppress organ-specific autoimmune diseases. The mechanism of CD25(+)CD4(+) T cells in the regulation of experimental autoimmune encephalomyelitis (EAE) is poorly understood. To assess the role of CD25(+)CD4(+) T cells in EAE, SJL mice were immunized with myelin proteolipid protein (PLP)(139 151) to develop EAE and were treated with anti-CD25 mAb. Treatment with anti-CD25 antibody following immunization resulted in a significant enhancement of EAE disease severity and mortality. There was increased inflammation in the central nervous system (CNS) of anti-CD25 mAb-treated mice. Anti-CD25 antibody treatment caused a decrease in the percentage of CD25(+)CD4(+) T cells in blood, peripheral lymph node (LN) and spleen associated with increased production of IFN-gamma and a decrease in IL-10 production by LN cells stimulated with PLP(130-151) in vitro. In addition, transfer of CD25(+)CD4(+) regulatory T cells from naive SJL mice decreased the severity of active EAE. In vitro, anti-CD3-stimulated CD25(+)CD4(+) T cells from naive SJL mice secreted IL-10 and IL-10 soluble receptor (sR) partially reversed the in vitro suppressive activity of CD25(+)CD4(+) T cells. CD25(+)CD4(+) T cells from IL-10-deficient mice were unable to suppress active EAE. These findings demonstrate that CD25(+)CD4(+) T cells suppress pathogenic autoreactive T cells in actively induced EAE and suggest they may play an important natural regulatory function in controlling CNS autoimmune disease through a mechanism that involves IL-10. PMID- 14734611 TI - Role of L-selectin in the development of autoimmune diabetes in non-obese diabetic mice. AB - Autoimmune diabetes is characterized by an early mononuclear infiltration of pancreatic islets and later selective autoimmune destruction of insulin-producing beta cells. Lymphocyte homing receptors have been considered candidate targets to prevent autoimmune diabetes. L-selectin (CD62L) is an adhesion molecule highly expressed in naive T and B cells. It has been reported that blocking L-selectin in vivo with a specific antibody (Mel-14) partially impairs insulitis and diabetes in autoimmune diabetes-prone non-obese diabetic (NOD) mice. In the present study we aimed to elucidate whether genetic blockade of leukocyte homing into peripheral lymph nodes would prevent the development of diabetes. We backcrossed L-selectin-deficient mice onto the NOD genetic background. Surprisingly NOD/L-selectin-deficient mice exhibited unaltered islet mononuclear infiltration, timing of diabetes onset and cumulative incidence of spontaneous diabetes when compared to L-selectin-sufficient animals. CD4, CD8 T cells and B cells were present in islet infiltrates from 9-week-old L-selectin-sufficient and -deficient littermates. Moreover, total splenocytes from wild-type, heterozygous or NOD/L-selectin-deficient donor mice showed similar capability to adoptively transfer diabetes into NOD/SCID recipients. On the other hand, homing of activated, cloned insulin-specific autoaggressive CD8 T cells (TGNFC8 clone) is not affected in NOD/L-selectin-deficient recipients. We conclude that L-selectin plays a small role in the homing of autoreactive lymphocytes to regional (pancreatic) lymph nodes in NOD mice. PMID- 14734612 TI - Molecular basis of Stat1 and PU.1 cooperation in cytokine-induced Fcgamma receptor I promoter activation. AB - The high-affinity receptor for IgG (FcgammaRI) is a myeloid cell-specific and IFN gamma-induced gene, and thereby serves as a paradigm for cytokine-induced cell type-specific gene responses. The expression of FcgammaRI is regulated by PU.1 and Stat1 transcription factors. We established an experimental model to analyze the individual functions of Stat1 and PU.1 in cytokine-induced transcription of the natural FcgammaRI promoter in U3A cells lacking both factors. PU.1 was required for both the basal activity and for the IFN-gamma-induced FcgammaRI promoter activation, while Stat1 alone could not initiate transcription. In contrast, in the context of a heterologous promoter, PU.1 inhibited the Stat1 mediated transcription. Systematic analysis of Stat1 and PU.1 mutants and FcgammaRI promoter elements revealed that activation of the promoter required the DNA binding, and the transactivation functions of both Stat1 and PU.1. PU.1 and Stat1 bound the promoter elements independently, and no physical interaction between the proteins was observed. The requirement of PU.1 for FcgammaRI promoter activity was supported by demonstration of in vitro interaction between PU.1 and components of the basal transcription machinery TBP and RNA polymerase II. Deletion of the acidic transactivation domain of PU.1 greatly diminished both the FcgammaRI promoter activity as well as the interaction with RNA polymerase II. In contrast, Stat1 did not interact with TBP or RNA polymerase II. These results define functional cooperativity between PU.1 and Stat1 in FcgammaRI promoter activation where PU.1 serves as an amplifier and bridging factor with the basal transcription machinery. PMID- 14734613 TI - Allicin stimulates lymphocytes and elicits an antitumor effect: a possible role of p21ras. AB - Allicin, the main organic allyl sulfur component in garlic, exhibits immune stimulatory and antitumor properties. Allicin stimulated [(3)H]thymidine incorporation in mouse splenocytes and enhanced cell-mediated cytotoxicity in human peripheral mononuclear cells. Multiple administration (i.p.) of allicin elicited a marked antitumor effect in mice inoculated with B-16 melanoma and MCA 105 fibrosarcoma. The immune-stimulatory and antitumor effects of allicin are characterized by a bell-shaped curve, i.e. allicin at high, supra-optimal concentrations is less effective or inhibitory. Allicin induced activation of extracellular signal-regulated kinases 1 and 2 (ERK1/2) in human peripheral mononuclear cells, and also in wild-type Jurkat T-cells. Allicin failed to activate ERK1/2 in Jurkat T cells that express p21(ras), in which Cys118 was replaced by Ser. These cells are not susceptible to redox-stress modification and activation. We postulate that the immune stimulatory effect of allicin is mediated by redox-sensitive signaling such as activation of p21(ras). It is suggested that the antitumor effect of allicin is related to its immune stimulatory properties. PMID- 14734614 TI - Analysis of autoreactive T cells associated with murine collagen-induced arthritis using peptide-MHC multimers. AB - CD4(+) T cells that recognize residues 256-270 of type II collagen (CII) associated with the I-A(q) (A(q)) molecule play a central role in disease pathogenesis in murine collagen-induced arthritis (CIA). Disease is most efficiently induced by immunization with heterologous CII, which elicits heterologous, e.g. bovine, CII256-270:I-A(q)-specific T cells that only poorly cross-react with mouse CII. The self-epitope differs from heterologous CII256-270 by a conservative change of glutamic acid (heterologous) to aspartic acid (mouse) at position 266 which confers a lower affinity for binding to the I-A(q) molecule. To date, characterization of the nature of T cell recognition in this model has been hindered by the lack of suitable, labeled multimeric peptide-MHC class II complexes. Here, we describe the biochemical properties of both recombinant bovine CII256-270:I-A(q) (bCII256-270:I-A(q)) and mouse CII256-270:I A(q) (mCII256-270:I-A(q)) complexes, and use these as fluorescently labeled multimers (tetramers) to characterize the specificity of CII-reactive T cells. Our analyses show that an unexpectedly high percentage of bCII256-270:I-A(q) specific T cells are cross-reactive with mCII256-270:I-A(q). Interestingly, one T cell clone which has a relatively high avidity for binding to self-CII256-270:I A(q) shows a marked increase in binding avidity at physiological temperature, indicating that this TCR has unusual thermodynamic properties. Taken together, our analyses suggest that the low affinity of mCII256-270 for I-A(q) may lead to a state of ignorance which can be overcome by priming CII-specific T cells with heterologous CII. This has relevance to understanding the mechanism by which CIA is induced and provides an explanation for the low arthritogenicity of mouse CII. PMID- 14734615 TI - A mechanism underlying STAT4-mediated up-regulation of IFN-gamma induction inTCR triggered T cells. AB - IL-12 promotes T(h)1 development/IFN-gamma expression by activating STAT4. However, it is still unclear how STAT4 elicits IFN-gamma promoter activation. Here, we investigated the mechanism by which IL-12-activated STAT4 functions for IFN-gamma induction in TCR-triggered T cells. TCR stimulation induced high levels of IFN-gamma production depending on co-stimulation with IL-12. IL-12 stimulation greatly enhanced the promoter-binding activity of c-Jun/AP-1, a critical transcription factor for IFN-gamma gene expression in wild-type T cells, but not in STAT4-deficient (STAT4(-/-)) T cells. Comparable amounts of c-Jun were induced by TCR stimulation in both wild-type and STAT4(-/-) T cells irrespective of IL-12 co-stimulation. However, c-Jun bound to STAT4 in IL-12-co-stimulated wild-type T cells. c-Jun forming a complex with STAT4 efficiently interacted with the AP-1 related sequence of the IFN-gamma promoter. Such an enhanced c-Jun binding did not occur in STAT4(-/-) T cells. These results show that STAT4 contributes to enhancing IFN-gamma expression by up-regulating the binding of TCR signal-induced AP-1 to the relevant promoter sequence. PMID- 14734616 TI - Inadequate induction of suppressor of cytokine signaling-1 causes systemic autoimmune diseases. AB - Suppressor of cytokine signaling (SOCS)-1 is a cytokine-inducible, negative regulatory molecule of Janus kinases (JAK) and its deficiency causes hyper response to various cytokines. SOCS-1(-/-) mice spontaneously develop a fatal disease depending on aberrantly activated lymphocytes. Here, we show that partial restoration of SOCS-1 in lymphoid cells rescues SOCS-1(-/-) mice from the early onset fatal disease, indicating that SOCS-1 expression in vivo is especially required in lymphocytes. However, SOCS-1 expression in these SOCS-1-restored mutant mice (E( micro )-SOCS-1(-/-) mice) was insufficient for proper down regulation of its target signaling, and these mice spontaneously exhibit hyperactivation of lymphocytes, an increase in the levels of serum Ig and anti DNA autoantibodies, and glomerulonephritis with glomerular IgG deposition. These phenotypes resemble those of murine systemic autoimmune diseases, models for systemic lupus erythematosus (SLE). Interestingly, similar phenotypes were also observed in adult female SOCS-1(+/-) mice, indicating that the autoimmune phenotypes of these mice can be ascribed primarily to the inadequate expression of SOCS-1. In addition, autoimmune phenotypes were not observed in SOCS-1(+/ )CD4(-/-) mice, suggesting that autoimmunity is dependent on hyper-activated CD4(+) T cells. Our findings also suggest that insufficient expression of SOCS-1 results in impaired function of CD25(+)CD4(+) regulatory T cells, which may contribute to aberrant activation of CD4(+) T cells. These findings suggest that dysfunction of SOCS-1 can be a pathogenic factor of systemic autoimmune diseases such as SLE. PMID- 14734617 TI - Immunologic immaturity, but high IL-4 productivity, of murine neonatal thymic CD4 single-positive T cells in the last stage of maturation. AB - To determine the levels of maturation and differentiation of murine CD4 single positive (SP) T cells, we compared the secondary responses of staphylococcal enterotoxin A (SEA)-induced neonatal thymic, adult thymic and adult splenic CD4 SP T cell blasts prepared from whole or heat-stable antigen(low) CD4 SP T cells. Proliferative responses upon re-stimulation with SEA were strong in adult splenic CD4 SP T cell blasts, but quite weak in neonatal thymic and adult thymic CD4 SP T cell blasts. SEA-induced IL-2 production was weaker in neonatal thymic blasts than in the adult splenic CD4 SP T cell blasts. In contrast, SEA-induced IL-4 production was high in neonatal thymic CD4 SP T cell blasts, and low in adult splenic and thymic CD4 SP T cell blasts. Expression of GATA-3, that directs production of IL-4 in T cells, examined at protein and mRNA levels, was higher in neonatal thymic cells than in adult thymic and splenic cells. These results suggest that neonatal and adult thymic CD4 SP T cells in the final stage of maturation are relatively immature compared with adult splenic CD4 SP T cells. The cytokine production profile of neonatal thymic CD4 SP T cells suggests that they are inclined towards a T(h)2 response. PMID- 14734618 TI - Incomplete tolerance to the tumour-associated antigen MDM2. AB - MDM2 is a tumour-associated antigen widely expressed by normal tissues and over expressed by many tumours of different origin. We wanted to define the level of immunological tolerance against MDM2 and explore its potential in tumour immunotherapy. Two murine MDM2 epitopes, pMDM100 and pMDM441, differ in their affinity for MHC class I molecules. Previous observations made in vitro suggested that pMDM100, due to its high affinity for K(b), induces a high level of tolerance, whereas tolerance to pMDM441, which binds poorly to D(b), is incomplete. In the present article we test the immunogenicity of these two peptides in vivo. Surprisingly, mice immunized with pMDM100 generated cytotoxic T lymphocytes (CTL) that killed tumour cell lines expressing MDM2 endogenously, indicating the existence of high-avidity CTL specific for a widely expressed protein. However, the response was limited as effector CTL disappeared after continued in vitro stimulation. While immunization with the individual MDM2 peptides did not protect against tumour challenge, mice immunized with both pMDM100 and pMDM441 were partially protected. These observations suggest that targeting of multiple epitopes may be required to vaccinate against tumours expressing elevated levels of CTL-recognized self-proteins. PMID- 14734619 TI - Roles of caspase-1 in Listeria infection in mice. AB - Caspase-1 [IL-1beta-converting enzyme (ICE)] processes substrate precursor molecules to yield the biologically active form of IL-1beta and IL-18, both of which are considered to play important roles in the host defense by activation of both innate and adaptive immunity. We evaluated the immune response of caspase-1( /-) mice to Listeria monocytogenes (LM) infection. LM eradication in the early phase of infection was impaired in the mutant mice with a prominent decrease in IL-18 and IFN-gamma production, but not in IL-12. Caspase-1(-/-) spleen cells including dendritic cells and NK cells produced less IFN-gamma in response to heat-killed LM than wild-type cells in vitro. IFN-gamma production and bactericidal activity in LM-infected caspase-1(-/-) mice was reconstituted to normal levels by adding back IL-18 at the initial phase of infection, suggesting that the lack of this cytokine is primarily responsible for the susceptibility of caspase-1(-/-) mice against LM infection. Moreover, IFN-gamma injection of caspase-1(-/-) mice corrected the deficiency in pathogen clearance. In contrast, LM-specific acquired immunity in caspase-1(-/-) mice was normal and they successfully cleared the pathogen following secondary infection, in spite of a moderate skewing of cytokine profile to T(h)2 when compared to wild-type mice. These data shed light on the importance of caspase-1-mediated IL-18 processing in innate immunity against facultative intracellular pathogens. PMID- 14734620 TI - Development of dermatitis in CD18-deficient PL/J mice is not dependent on bacterial flora, and requires both CD4+ and CD8+ T lymphocytes. AB - CD18-deficient PL/J mice develop dermatitis characterized by hyperkeratosis, and a mixed dermal and epidermal inflammatory infiltrate. The development of this disease requires low-level CD18 expression and at least two PL/J loci. Currently, the mechanisms by which decreased beta(2) integrin expression on leukocytes promotes skin inflammation in PL/J mice are unknown. In these studies, we investigated the role of microbial infection and T lymphocytes in the pathogenesis of this disease. We found that germ-free CD18(-/-) PL/J mice developed dermatitis indistinguishable from that of mice raised in pathogen-free conditions. Adoptive transfer of CD18(-/-) PL/J splenocytes into skin disease resistant CD18(+/-) PL/J mice failed to induce skin inflammation. However, transfer of CD18(+/-) splenocytes blocked the progression and ultimately led to resolution of skin disease in the majority of CD18(-/-) recipients. Depletion of both CD4(+) and CD8(+) T cells mice prior to onset of the disease significantly delayed the appearance of inflammatory skin disease. In contrast, single depletions of these T cells did not inhibit disease development. These studies show that dermatitis in CD18-deficient PL/J mice is not the consequence of infection, does not require bacterial superantigens, and is mediated by both CD4(+) and CD8(+) T lymphocytes. Furthermore, they suggest that one possible mechanism for skin disease development in these mice may involve the absence or dysfunctional activity of a regulatory T cell population. These mice may therefore be useful in identifying potential mechanisms of pathogenesis and genetic predisposition in human inflammatory skin diseases. PMID- 14734621 TI - Microvilli structures on B lymphocytes: inducible functional domains? AB - Interactive contact between B lymphocytes and T cells is necessary for their expansion during an immune response. It has been shown that B lymphocytes receive signals from T cells, such as IL-4 and cross-linking of CD40, which are crucial for their differentiation. We previously found that these factors induce formation of microvilli on B cells and that this was correlated with increased homotypic adhesion of B lymphocytes. In this study we have investigated if IL-4 induce segregation of proteins to microvilli and lipid rafts. Using immuno electron microscopy we analyzed cell-surface distribution of molecules involved in B-T cell co-activation. Recruitment to detergent-resistant membrane fractions was analyzed using sucrose gradient centrifugation. We found that microvilli were enriched in ICAM-1 and MHC class II molecules. In contrast, LFA-1 and CD40 were more abundant on the smooth cell surfaces, while B7-2 (CD86) was randomly distributed. We also discovered that depletion of cholesterol, using beta-methyl cyclodextrin, lowered the number of microvilli, indicating that intact lipid rafts are required for their expression. Moreover, activation of B lymphocytes by lipopolysaccharide (LPS) induced increased expression of GM(1), a marker for lipid rafts. However, although both surface and total levels of GM(1) were similar in B lymphocytes stimulated with either LPS or LPS plus IL-4, GM(1) was mainly expressed on microvilli in LPS plus IL-4-stimulated cells. Taken together, our results indicate that microvilli represent distinct inducible membrane domains that can regulate direct cell-cell interactions via grouping and three dimensional presentation of cell-surface receptors. PMID- 14734622 TI - Phenotypic characterization of regulatory CD4+CD25+ T cells in rats. AB - CD25 has become widely used as a marker for a subset of regulatory CD4(+) T cells present in the thymus and periphery of mice, rats and humans. However, CD25 is also expressed on conventionally activated T cells that are not regulatory and not all peripheral regulatory T cells express CD25. The identification of a stable and unique marker for regulatory T cells would therefore be valuable. This study provides a detailed account of the phenotype of CD4(+)CD25(+) regulatory T cells in rats. In the thymus, CD4(+)CD8(-)CD25(+) cells were found to have a more mature phenotype than the corresponding CD4(+)CD8(-)CD25(-) cells with respect to expression of Thy1 (CD90), CD53 and CD44, suggesting that CD25 expression, and perhaps commitment to regulatory function, might be a late event in thymocyte development. CD4(+)CD25(+) cells in both the thymus and periphery were found to have enriched and heterogeneous expression of activation markers such as OX40 (CD134) and OX48 (an antibody determined in this study to be specific for CD86). CD4(+)CD25(+) T cells were also found to have enriched expression of CD80, at both the mRNA and protein level. However, functional studies in vitro and in vivo showed that neither OX40 or CD86 were useful markers for the further subdivision of regulatory T cells. Our studies indicate that, at present, CD25 remains the most useful marker to enrich for regulatory CD4(+) T cells in rats and no further subdivision of the regulatory component of CD4(+)CD25(-)CD45RC(low) T cells has yet been achieved. PMID- 14734623 TI - Germinal center checkpoints in B cell tolerance in 3H9 transgenic mice. AB - Regulation throughout B cell maturation and activation prevents autoreactive B cells from entering germinal center (GC) reactions. This study shows that a subset of autoreactive B cells in V(H)3H9 micro IgH transgenic mice escapes these serial checkpoints and proceeds into splenic GC. GC B cells isolated from these mice all express the transgenic V(H)3H9 micro heavy chain, some co-express light chains that yield an anti-dsDNA specificity and some have somatic mutations, consistent with their GC origin. Nonetheless, B cell tolerance is ultimately preserved as serum titers of anti-dsDNA antibodies are not elevated. These observations suggest that those autoreactive GC B cells that escaped earlier checkpoints and possibly also those cells that acquire autoreactivity de novo by mutating their antigen receptor are arrested within the splenic GC before differentiating further into antibody-secreting plasma cells. PMID- 14734624 TI - The impact of SNP density on fine-scale patterns of linkage disequilibrium. AB - Linkage disequilibrium (LD) is a measure of the degree of association between alleles in a population. The detection of disease-causing variants by association with neighbouring single nucleotide polymorphisms (SNPs) depends on the existence of strong LD between them. Previous studies have indicated that the extent of LD is highly variable in different chromosome regions and different populations, demonstrating the importance of genome-wide accurate measurement of LD at high resolution throughout the human genome. A uniform feature of these studies has been the inability to detect LD in regions of low marker density. To investigate the dependence of LD patterns on marker selection we performed a high-resolution study in African-American, Asian and UK Caucasian populations. We selected over 5000 SNPs with an average spacing of approximately 1 SNP per 2 kb after validating ca 12 000 SNPs derived from a dense SNP collection (1 SNP per 0.3 kb on average). Applications of different statistical methods of LD assessment highlight similar areas of high and low LD. However, at high resolution, features such as overall sequence coverage in LD blocks and block boundaries vary substantially with respect to marker density. Model-based linkage disequilibrium unit (LDU) maps appear robust to marker density and consistently influenced by marker allele frequency. The results suggest that very dense marker sets will be required to yield stable views of fine-scale LD in the human genome. PMID- 14734625 TI - Coordinated replication timing of monoallelically expressed genes along human autosomes. AB - A number of genes in the mammalian genome are expressed from only one of two alleles in either an imprinted or random manner. Those belonging to the random class include X-linked genes subject to X inactivation, as well as a number of autosomal genes, including odorant receptors, immunoglobulins, T-cell receptors, interleukins, natural killer-cell receptors and pheromone receptors. Monoallelically expressed genes display the unusual property of asynchronous replication and for those genes whose transcription is randomly monoallelic, the asynchronous replication is also random. In mice, recent work has shown that the random asynchronous replication of distributed autosomal genes is coordinated at the whole chromosome level, indicative of chromosome-pair non-equivalence. Here, we show that autosome-pair non-equivalence is present in human cells, and demonstrate its ability to cross the centromere. Additionally, by examining the replication of these genes in a number of human trisomies, we consistently find one allele replicating early and the other two alleles replicating late, similar to previous observations in X trisomies. PMID- 14734626 TI - Multiple pathways regulate MeCP2 expression in normal brain development and exhibit defects in autism-spectrum disorders. AB - Rett syndrome (RTT) is a neurodevelopmental disorder caused by mutations in MECP2, encoding methyl-CpG-binding protein 2 (MeCP2). Although MECP2 is ubiquitously transcribed, MeCP2 expression is developmentally regulated and heterogeneous in neuronal subpopulations, defined as MeCP2(lo) and MeCP2(hi). To test the hypothesis that pathways affecting MeCP2 expression changes may be defective in RTT, autism and other neurodevelopmental disorders without MECP2 mutations, a high-throughput quantitation of MeCP2 expression was performed on a tissue microarray containing frontal cortex samples from 28 different patients with neurodevelopmental disorders and age-matched controls. Combined quantitative analyses of MeCP2 protein and alternatively polyadenylated transcript levels were performed by laser scanning cytometry and tested for significant differences from age-matched controls. Normal cerebral samples showed an increase in total MeCP2 expression and the percentage of MeCP2(hi) cells with age that could be explained by increased MECP2 transcription within the MeCP2(hi) population. A significant decrease in the relative usage of the long transcript in the MeCP2(lo) population was observed in postnatal compared to fetal brain, but alternate polyadenylation did not correlate with MeCP2 expression changes at the single cell level. Brain samples from several related neurodevelopmental disorders, including autism, pervasive developmental disorder, Prader-Willi and Angelman syndromes showed significant differences in MeCP2 expression from age-matched controls by apparently different transcriptional and post-transcriptional mechanisms. These results suggest that multiple pathways regulate the complex developmental expression of MeCP2 and are defective in autism-spectrum disorders in addition to RTT. PMID- 14734627 TI - Inhibition of myogenesis in transgenic mice expressing the human DMPK 3'-UTR. AB - Myotonic dystrophy (DM1) is a multisystemic disorder caused by a CTG repeat expansion within the 3'-UTR of the DMPK gene. DM1 is characterized by delayed muscle development, muscle weakness and wasting, cardiac conduction abnormalities, cognitive defects and cataracts. Recent studies have demonstrated that the disease mechanism involves a dominant gain-of-function conferred upon mutant transcripts by expanded repeats. However, further attempts to model aspects of DM muscle pathology in cultured myoblasts suggest that 3'-UTR sequences flanking the CTG repeat tract are also required for full expression of the disease phenotype. Here, we report that overexpression of the DMPK 3'-UTR including either wild-type (11) or expanded (91) CTG repeats results in aberrant and delayed muscle development in fetal transgenic mice. In addition, transgenic animals with both expanded and wild-type CTG repeats display muscle atrophy at 3 months of age. Primary myoblast cultures from both 11 and 91 repeat mice display reduced fusion potential, but a greater reduction is observed in the 91 repeat cultures. Taken together, these data indicate that overexpression of the DMPK 3' UTR interferes with normal muscle development in mice and that this is exacerbated by inclusion of a mutant repeat. This suggests that the delayed muscle development in DM1 involves an interplay between the expanded CTG repeat and adjacent 3'-UTR sequences. PMID- 14734628 TI - Systematic changes in gene expression in postmortem human brains associated with tissue pH and terminal medical conditions. AB - Studies of gene expression abnormalities in psychiatric or neurological disorders often involve the use of postmortem brain tissue. Compared with single-cell organisms or clonal cell lines, the biological environment and medical history of human subjects cannot be controlled, and are often difficult to document fully. The chance of finding significant and replicable changes depends on the nature and magnitude of the observed variations among the studied subjects. During an analysis of gene expression changes in mood disorders, we observed a remarkable degree of natural variation among 120 samples, which represented three brain regions in 40 subjects. Most of such diversity can be accounted for by two distinct expression patterns, which in turn are strongly correlated with tissue pH. Individuals who suffered prolonged agonal states, such as with respiratory arrest, multi-organ failure or coma, tended to have lower pH in the brain; whereas those who experienced brief deaths, associated with accidents, cardiac events or asphyxia, generally had normal pH. The lower pH samples exhibited a systematic decrease in expression of genes involved in energy metabolism and proteolytic activities, and a consistent increase of genes encoding stress response proteins and transcription factors. This functional specificity of changed genes suggests that the difference is not merely due to random RNA degradation in low pH samples; rather it reflects a broad and actively coordinated biological response in living cells. These findings shed light on critical molecular mechanisms that are engaged during different forms of terminal stress, and may suggest clinical targets of protection or restoration. PMID- 14734629 TI - Type 2 diabetes mellitus: not quite exciting enough? AB - Type 2 diabetes mellitus is a serious metabolic disease that afflicts around 5% of the population in Western societies and over 150 million people worldwide. It is characterized by elevation of the blood glucose concentration, usually presents in middle age, and is exacerbated by obesity. Both genetic and environmental factors contribute to the disease but in the vast majority of cases the aetiology is still not understood. Here we present a novel hypothesis for the aetiology of type 2 diabetes. We postulate that the electrical activity of the insulin-secreting beta-cells of the pancreas acts to integrate the genetic and environmental factors that predispose to disease risk. Our hypothesis is supported by a substantial amount of data gathered from a range of different disciplines and makes predictions that can be tested experimentally both in vitro and in man. PMID- 14734630 TI - New aspects of altitude adaptation in Tibetans: a proteomic approach. AB - A prolonged sojourn above 5500 m induces muscle deterioration and accumulation of lipofuscin in Caucasians, probably because of overproduction of reactive oxygen species (ROS). Because Sherpas, who live at high altitude, have very limited muscle damage, it was hypothesized that Himalayan natives possess intrinsic mechanisms protecting them from oxidative damage. This possibility was investigated by comparing the muscle proteomes of native Tibetans permanently residing at high altitude, second-generation Tibetans born and living at low altitude, and Nepali control subjects permanently residing at low altitude, using 2D gel electrophoresis and mass spectrometry. Seven differentially regulated proteins were identified: glutathione-S-transferase P1-1, which was 380% and 50% overexpressed in Tibetans born and living at high and low altitude, respectively; Delta2-enoyl-CoA-hydratase, which was up-regulated in both Tibetan groups; glyceraldehyde-3-phosphate dehydrogenase and lactate dehydrogenase, which were both slightly down-regulated in Tibetans born and living at high altitude; phosphoglycerate mutase, which was 50% up-regulated in the native Tibetans; NADH ubiquinone oxidoreductase, slightly overexpressed in Tibetans born and living at high altitude; and myoglobin, which was overexpressed in both Tibetan groups. We concluded that Tibetans at high altitude, and to some extent, those born and living at low altitude, are protected from ROS-induced tissue damage and possess specific metabolic adaptations. PMID- 14734631 TI - Hydrogen sulfide induces calcium waves in astrocytes. AB - Hydrogen sulfide (H2S) modifies hippocampal long-term potentiation (LTP) and functions as a neuromodulator. Here, we show that H2S increases intracellular Ca2+ and induces Ca2+ waves in primary cultures of astrocytes as well as hippocampal slices. H2S increases the influx of Ca2+ and to a lesser extent causes the release from intracellular Ca2+ stores. Ca2+ waves induced by neuronal excitation as well as responses to exogenously applied H2S are potently blocked by La3+ and Gd3+, inhibitors of Ca2+ channels. These observations suggest that H2S induces Ca2+ waves that propagate to neighboring astrocytes. PMID- 14734632 TI - Neuroprotective effect of dextromethorphan in the MPTP Parkinson's disease model: role of NADPH oxidase. AB - Parkinson's disease (PD) is a neurodegenerative movement disorder characterized by a progressive loss of dopaminergic neurons in the substantia nigra and depletion of the neurotransmitter dopamine in the striatum. Progress in the search for effective therapeutic strategies that can halt this degenerative process remains limited. Mechanistic studies using animal systems such as the 1 methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) rodent PD model have revealed the involvement of the brain's immune cells and free radical-generating processes. We recently reported that dextromethorphan (DM), a widely used anti tussive agent, attenuated endotoxin-induced dopaminergic neurodegeneration in vitro. In the current study, we investigated the potential neuroprotective effect of DM and the underlying mechanism of action in the MPTP rodent PD model. Mice (C57BL/6J) that received daily MPTP injections (15 mg free base/kg body weight, s.c.) for 6 consecutive days exhibited significant degeneration of the nigrostriatal dopaminergic pathway. However, the MPTP-induced loss of nigral dopaminergic neurons was significantly attenuated in those mice receiving DM (10 mg/kg body weight, s.c.). In mesencephalic neuron-glia cultures, DM significantly reduced the MPTP-induced production of both extracellular superoxide free radicals and intracellular reactive oxygen species (ROS). Because NADPH oxidase is the primary source of extracellular superoxide and intracellular ROS, we investigated the involvement of NADPH oxidase in the neuroprotective effect of DM. Indeed, the neuroprotective effect of DM was only observed in the wild-type but not in the NADPH oxidase-deficient mice, indicating that NADPH oxidase is a critical mediator of the neuroprotective activity of DM. More importantly, due to its proven safety record of long-term clinical use in humans, DM may be a promising agent for the treatment of degenerative neurological disorders such as PD. PMID- 14734633 TI - Plexin signaling hampers integrin-based adhesion, leading to Rho-kinase independent cell rounding, and inhibiting lamellipodia extension and cell motility. AB - Plexins encode receptors for semaphorins, molecular signals guiding cell migration, and axon pathfinding. The mechanisms mediating plexin function are poorly understood. Plexin activation in adhering cells rapidly leads to retraction of cellular processes and cell rounding "cell collapse"). Here we show that, unexpectedly, this response does not require the activity of Rho-dependent kinase (ROCK) nor the contraction of F-actin cables. Interestingly, integrin based focal adhesive structures are disassembled within minutes upon plexin activation; this is followed by actin depolymerization and, eventually, by cellular collapse. We also show that plexin activation hinders cell attachment to adhesive substrates, blocks the extension of lamellipodia, and thereby inhibits cell migration. We conclude that plexin signaling uncouples cell substrate adhesion from cytoskeletal dynamics required for cell migration and axon extension. PMID- 14734634 TI - Chromogranin A protects vessels against tumor necrosis factor alpha-induced vascular leakage. AB - Elevated levels of circulating chromogranin A (CgA), a protein stored in the secretory granules of many neuroendocrine cells and neurons, have been detected in the blood of patients with neuroendocrine tumors or heart failure. The pathophysiological role of increased secretion of CgA is unknown. Using mice bearing subcutaneous tumors genetically engineered to secrete CgA in circulation, we have found that increased blood levels of this protein prevent vascular leakage induced by tumor necrosis factor-alpha (TNF) in the liver venous system. Structure-activity studies, carried out with CgA fragments administered to normal mice, showed that an active site is located within residues 7-57 of CgA. Accordingly, an anti-CgA antibody directed to residues 53-57 inhibited the effect of circulating CgA, either endogenously produced or exogenously administered, on liver vessels. Studies of the mechanism of action showed that CgA inhibits TNF induced VE-cadherin down-regulation and barrier alteration of cultured endothelial cells, in an indirect manner. Other effectors, such as thrombin and vascular endothelial growth factor were partially inhibited by CgA N-terminal fragments in in vitro permeability assays. These findings suggest that circulating CgA could help regulate the endothelial barrier function and to protect vessels against TNF-induced plasma leakage in pathological conditions characterized by increased production of TNF and CgA, such as cancer or heart failure. PMID- 14734635 TI - Mitochondrial and nuclear DNA base excision repair are affected differently by caloric restriction. AB - Aging is strongly correlated with the accumulation of oxidative damage in DNA, particularly in mitochondria. Oxidative damage to both mitochondrial and nuclear DNA is repaired by the base excision repair (BER) pathway. The "mitochondrial theory of aging" suggests that aging results from declining mitochondrial function, due to high loads of damage and mutation in mitochondrial DNA (mtDNA). Restriction of caloric intake is the only intervention so far proven to slow the aging rate. However, the molecular mechanisms underlying such effects are still unclear. We used caloric-restricted (CR) mice to investigate whether lifespan extension is associated with changes in mitochondrial BER activities. Mice were divided into two groups, receiving 100% (PF) or 60% (CR) of normal caloric intake, a regime that extends mean lifespan by approximately 40% in CR mice. Mitochondria isolated from CR mice had slightly higher uracil (UDG) and oxoguanine DNA glycosylase (OGG1) activities but marginally lower abasic endonuclease and polymerase gamma gap-filling activities, although these differences were tissue-specific. Uracil-initiated BER synthesis incorporation activities were significantly lower in brain and kidney from CR mice but marginally enhanced in liver. However, nuclear repair synthesis activities were increased by CR, indicating differential regulation of BER in the two compartments. The results indicate that a general up-regulation of mitochondrial BER does not occur in CR. PMID- 14734636 TI - SING: a novel strategy for identifying tumor-specific, CTL-recognized tumor antigens. AB - Traditional methods for identifying T cell-recognized tumor antigens (Ags) are laborious and time-consuming. In an attempt to simplify the procedure, a novel strategy, SING (SIgnal transduction molecule-mediated, NFAT-controlled, GFP expression) was established as a direct approach for cloning T cell-recognized tumor Ags. In the SING system, a mouse T cell line (BW5147) was transduced with a chimeric H-2Kb construct containing T cell-signaling domains and a green fluorescent protein (GFP) reporter gene under the transcriptional control of nuclear factor of activated T cells (NFAT). The resultant BW5147 cells were named BS cells. This cell line could "sense" TCR stimulation through the T cell signaling domains after coculture with Ag-specific T cells and then become fluorescent (expressing green fluorescence protein, GFP+) in the presence of Ag peptides. The interaction between BS cells and Ag-specific T cells could be enhanced by addition of costimulatory signals. Currently, BS cells have been optimized to "sense" TCR stimulation after being pulsed with the relevant peptides at concentrations as low as 10(-9) M. Endogenous Ag-expressing BS cells could also become fluorescent after coculture with Ag-specific T cells. Our results provide a proof of principle for using the SING system to directly isolate Ag-expressing BS cells from BS cell repertoires, which are retrovirally transduced with tumor-derived cDNA libraries. Once tumor Ag-marked BS cells are identified, the sequences encoding tumor Ags could be easily retrieved by PCR amplification of the genomic DNA using vector-specific primers. PMID- 14734637 TI - A nitric oxide-releasing derivative of enalapril, NCX 899, prevents progressive cardiac dysfunction and remodeling in hamsters with heart failure. AB - Nitric oxide (NO) production is known to be impaired in heart failure. A new compound (NCX 899), a NO-releasing derivative of enalapril was characterized, and its actions were evaluated in Bio 14.6 cardiomyopathic (CM) hamsters with heart failure. The hamsters were randomized to oral treatment for 4 weeks with vehicle (n=11), NCX 899 (NCX, 25 mg/kg, n=10), or enalapril (25 mg/kg, n=10). In the vehicle group, fractional shortening by echocardiography decreased (-23.6+/-2.0%) and LV end-diastolic dimension) increased (+10.9+/-1.0%), whereas fractional shortening increased (+17.5+/-4.4%) in NCX and was unchanged in the enalapril group (both P<0.01 vs. vehicle). End-diastolic dimension decreased only in NCX. LV contractility (LVdP/dt max and Emax) was significantly greater in NCX than in enalapril or vehicle, while relaxation (Tau) was shortened in both NCX and enalapril vs. vehicle. ACE activity was inhibited equally by NCX and enalapril in the CM hamster, and plasma nitrate levels were increased only in NCX (P<0.05 vs. enalapril and vehicle). In aortic strips endothelium-independent relaxation occurred only with NCX. The superior effects of NO-releasing enalapril (NCX) vs. enalapril alone to enhance vascular effects, increase LV contractility and prevent unfavorable remodeling and are consistent with vascular delivery of exogenous NO. NCX 899 may hold promise for the future treatment of heart failure. PMID- 14734638 TI - Alpha-syntrophin deletion removes the perivascular but not endothelial pool of aquaporin-4 at the blood-brain barrier and delays the development of brain edema in an experimental model of acute hyponatremia. AB - The formation of brain edema, commonly occurring as a potentially lethal complication of acute hyponatremia, is delayed following knockout of the water channel aquaporin-4 (AQP4). Here we show by high-resolution immunogold analysis of the blood-brain-barrier that AQP4 is expressed in brain endothelial cells as well as in the perivascular membranes of astrocyte endfeet. A selective removal of perivascular AQP4 by alpha-syntrophin deletion delays the buildup of brain edema (assessed by Diffusion-weighted MRI) following water intoxication, despite the presence of a normal complement of endothelial AQP4. This indicates that the perivascular membrane domain, which is peripheral to the endothelial blood-brain barrier, may control the rate of osmotically driven water entry. This study is also the first to demonstrate that the time course of edema development differs among brain regions, probably reflecting differences in aquaporin-4 distribution. The resolution of the molecular basis and subcellular site of osmotically driven brain water uptake should help design new therapies for acute brain edema. PMID- 14734639 TI - Overexpression of the small mitochondrial Hsp22 extends Drosophila life span and increases resistance to oxidative stress. AB - Heat shock proteins (Hsp) are involved in protein folding, transport and stress resistance. Studies reporting an increased mRNA level of hsp genes in aged Drosophila suggest that expression of Hsp might be beneficial in preventing damages induced by aging. Because oxidative damage is often observed in aged organisms and mitochondria are sensitive to reactive oxygen species, we tested the hypothesis that increased levels of a small Hsp localized in mitochondria, Hsp22 of Drosophila melanogaster, could protect mitochondrial proteins and influence the aging process. We demonstrate that a ubiquitous or a targeted expression of Hsp22 within motorneurons increases the mean life span by more than 30%. Hsp22 shows beneficial effects on early-aging events since the premortality phase displays the same increase as the mean lifespan. Moreover, flies expressing Hsp22 in their motorneurons maintain their locomotor activity longer as assessed by a negative geotaxis assay. The motorneurons-targeted expression of Hsp22 also significantly increases flies' resistance to oxidative injuries induced by paraquat (up to 35%) and thermal stress (39% at 30 degrees C and 23% at 37 degrees C). These observations establish Hsp22 as a key player in cell-protection mechanisms against oxidative injuries and aging in Drosophila and corroborate the pivotal role of mitochondria in the process of aging. PMID- 14734640 TI - Interferon-gamma plays a nonredundant role in mediating T cell-dependent outward vascular remodeling of allogeneic human coronary arteries. AB - Vascular remodeling (change in vessel diameter) rather than intimal hyperplasia is the most important predictor of luminal loss in immune-mediated arterial injury, yet little is known about its mechanisms. Here, we show that outward vascular remodeling and intimal thickening, two manifestations of arteriosclerosis with opposing effects on luminal size, result from immune effector mechanisms that are T-cell dependent and interferon (IFN)-gamma mediated. In our in vivo model of human coronary artery injury by allogeneic peripheral blood mononuclear cells, both processes occur concurrently and are characterized by T-cell infiltrates with a predominantly IFN-gamma-producing cytokine profile. Neutralization of IFN-gamma inhibits the arterial and intimal expansion, whereas administration of IFN-gamma enhances these effects. The nonredundant role of IFN-gamma in T-cell-dependent remodeling of human coronary arteries demonstrated here presents a new therapeutic target for preservation of vessel lumen in arteriosclerosis. PMID- 14734641 TI - Calorie restriction limits the generation but not the progression of mitochondrial abnormalities in aging skeletal muscle. AB - The effect of early-onset calorie restriction and aging on the accumulation of electron transport system (ETS) abnormalities was studied in rat skeletal muscle. Rectus femoris and vastus lateralis muscle fibers were analyzed for cytochrome c oxidase (COX) and succinate dehydrogenase (SDH) enzyme activities. Fibers displaying COX negative and SDH hyper reactive (COX-/SDH++) phenotype were followed through 1000-2000 micrometers to determine the frequency and length of these abnormalities as well as the physiological impact on fiber structure. Calorie restricted rats had fewer ETS abnormal muscle fibers. The mean length of ETS abnormal regions in ad libitum rat muscle fibers was similar to calorie restricted rat muscles. ETS abnormal fibers from both diet groups exhibited intra fiber atrophy. A negative correlation between ETS abnormality length and fiber cross-sectional area (CSA) ratio was observed in both ad libitum and calorie- restricted rats. Although calorie restriction reduced the number of ETS abnormalities, it did not affect the length or associated fiber atrophy of ETS abnormal regions once the abnormality was established. Thus, calorie restriction affects the onset but not the progression of electron transport system abnormalities, thereby, limiting a process that ultimately results in fiber breakage and fiber loss. PMID- 14734642 TI - Early-onset calorie restriction conserves fiber number in aging rat skeletal muscle. AB - The purpose of this work was to determine the effect of early-onset calorie restriction on sarcopenia in the aging rat. Ad libitum (AL) fed animals were examined at 5, 18, 21, and 36 months of age. Calorie-restricted (CR) rats, 40% restricted since 4 months of age, were examined at 21 and 36 months of age. By 36 months, vastus lateralis, rectus femoris and soleus muscles, from AL-fed rats, had significant muscle mass and fiber loss, and reduced muscle cross-sectional area. Mean fiber diameter decreased with age in the vastus lateralis and rectus femoris but not the soleus of AL-fed rats. The number of Type I fibers significantly increased in the vastus lateralis with age. Calorie restriction did not prevent muscle mass loss with age; however, it significantly reduced muscle mass loss between 21 and 36 months of age compared with age-matched AL cohorts. Calorie restriction prevented fiber loss with age, and this conservation of fiber number reduced muscle mass loss with age. PMID- 14734643 TI - Activation of toll-like receptor-9 induces progression of renal disease in MRL Fas(lpr) mice. AB - How bacterial or viral infections trigger flares of autoimmunity is poorly understood. As toll-like receptor (TLR)-9 activation by exogenous or endogenous CpG-DNA may contribute to disease activity of systemic lupus erythematosus, we examined the effects of CpG-oligodeoxynucleotides (ODN) or DNA derived from Escherichia coli (E. coli) on the course of nephritis in MRL(lpr/lpr) mice. In kidneys of these mice, TLR9 localized to glomerular, tubulointerstitial, and perivascular infiltrates. After intraperitoneal injection labeled CpG-ODN localized to glomerular and interstitial macrophages and dendritic cells in nephritic kidneys of MRL(lpr/lpr) mice but not in healthy MRL controls. Furthermore, murine J774 macrophages and splenocytes from MRL(lpr/lpr) mice, but not tubular epithelial cells, renal fibroblasts, or mesangial cells, expressed TLR9 and up-regulated CCL5/RANTES mRNA upon stimulation with CpG-ODN in vitro. In vivo both E. coli DNA and CpG-ODN increased serum DNA autoantibodies of the IgG2a isotype in MRL(lpr/lpr) mice. This was associated with progression of mild to crescentic glomerulonephritis, interstitial fibrosis, and heavy proteinuria. CpG ODN increased renal CCL2/MCP-1 and CCL5/RANTES expression associated with increased glomerular and interstitial leukocyte recruitment. In contrast control GpC-ODN had no effect. We conclude that TLR9 activation triggers disease activity of systemic autoimmunity, for example, lupus nephritis, and that adaptive and innate immune mechanisms contribute to the CpG-DNA-induced progression of lupus nephritis. PMID- 14734644 TI - Taurine transporter knockout depletes muscle taurine levels and results in severe skeletal muscle impairment but leaves cardiac function uncompromised. AB - Taurine is the most abundant free amino acid in heart and skeletal muscle. In the present study, the effects of hereditary taurine deficiency on muscle function were examined in taurine transporter knockout (taut-/-) mice. These mice show an almost complete depletion of heart and skeletal muscle taurine levels. Treadmill experiments demonstrated that total exercise capacity of taut-/- mice was reduced by >80% compared with wild-type controls. The decreased performance of taut-/- mice correlated with increased lactate levels in serum during exercise. Surprisingly, cardiac function of taut-/- mice as assessed by magnetic resonance imaging, echocardiography, and isolated heart studies showed a largely normal phenotype under both control and stimulated conditions. However, analysis of taut /- skeletal muscle revealed electromyographic abnormalities. (1)H nuclear magnetic resonance spectroscopy of tissue extracts showed that in the heart of taut-/- mice the lack of taurine was compensated by the up-regulation of various organic solutes. In contrast, a deficit of >10 mM in total organic osmolyte concentration was found in skeletal muscle. The present study identifies taurine transport as a crucial factor for the maintenance of skeletal muscle function and total exercise capacity, while cardiac muscle apparently can compensate for the loss of taurine. PMID- 14734645 TI - Apolipoprotein A-1 interaction with plasma membrane lipid rafts controls cholesterol export from macrophages. AB - Cholesterol efflux to apolipoprotein A-1 (apoA-1) from cholesterol-loaded macrophages is an important anti-atherosclerotic mechanism in reverse cholesterol transport. We recently provided kinetic evidence for two distinct pathways for cholesterol efflux to apoA-1 [Gaus et al. (2001) Biochemistry 40, 9363]. Cholesterol efflux from two membrane pools occurs sequentially with different kinetics; a small pool rapidly effluxed over the first hour, followed by progressive release from a major, slow efflux pool over several hours. In the present study, we propose that the rapid and slow cholesterol efflux pools represent cholesterol derived from lipid raft and nonraft domains of the plasma membrane, respectively. We provide direct evidence that apoA-1 binds to both lipid raft and nonraft domains of the macrophage plasma membrane. Conditions that selectively deplete plasma membrane lipid raft cholesterol, such as incorporation of 7-ketocholesterol or rapid exposure to cyclodextrins, block apoA-1 binding to these domains but also inhibit cholesterol efflux from the major, slow pool. We propose that cholesterol exported to apoA-1 from this major slow efflux pool derives from nonraft regions of the plasma membrane but that the interaction of apoA-1 with lipid rafts is necessary to stimulate this efflux. PMID- 14734646 TI - The gammaherpesvirus chemokine binding protein can inhibit the interaction of chemokines with glycosaminoglycans. AB - Chemokines are small glycosaminoglycan (GAG) binding proteins that direct the migration of leukocytes by signaling through G protein coupled receptors (GPCR). Many viruses encode proteins that disrupt chemokine responses. The murine gammaherpesvirus-68 gene M3 encodes a chemokine binding protein (vCKBP-3), which has no sequence similarity to chemokine receptors. Initial characterization of vCKBP-3 showed that it inhibits receptor binding and chemokine-induced calcium influx. The structural requirements for the chemokines CXCL8 and CCL2 to bind to vCKBP-3 have been determined. Both chemokines bind to vCKBP-3 via their N-loop, a site that can participate in GAG binding for some chemokines. We have investigated the effect of vCKBP-3 on the interaction of chemokines with GAGs. We found that vCKBP-3 can prevent a range of chemokines from binding to GAGs. Moreover, we also found that vCKBP-3 can displace chemokines from a heparin coated surface. Together, these data imply that vCKBP-3 can inhibit chemokine activity at two distinct levels. First, it inhibits chemokines from binding to their GPCR. Second, it inhibits their GAG binding and disrupts pre-formed chemokine gradients. This dual ability of vCKBP-3 makes it a more effective inhibitor of chemokine activity. PMID- 14734647 TI - Plasmin-activated doxorubicin prodrugs containing a spacer reduce tumor growth and angiogenesis without systemic toxicity. AB - To generate doxorubicin (Dox) specifically at the tumor site, the chemotherapeutic agent was incorporated into a prodrug by linkage to a peptide specifically recognized by plasmin, which is overproduced in many cancers. ST 9905, which contains an elongated self-elimination spacer, is activated more rapidly in vitro by plasmin than is ST-9802. Prodrug activation in vitro depended on the level of urokinase produced by tumor cells and was inhibited by aprotinin, a plasmin inhibitor. Comparison of equimolar concentrations of ST-9905, ST-9802, and Dox in EF43.fgf-4 and MCF7 models revealed that both prodrugs, in sharp contrast to Dox, displayed antiproliferative and antiangiogenic activities without discernible toxicity. Although MCF7 cells are poor urokinase producers in vitro, prodrug efficacy in this model may be explained by production of plasmin by tumor-infiltrating host cells. Mice treated with equitoxic concentrations (maximum tolerated doses) of prodrugs showed 100% survival and negligible body weight loss, in contrast to results after Dox treatment. ST-9905 was substantially more effective than ST-9802 and induced similar tumor growth inhibition as Dox but without apparent toxicity. This finding may be explained by the elongated spacer, which facilitates enzymatic prodrug activation. These data validate both the use of elongated spacers in vivo and the concept of targeting anticancer prodrugs to tumor-associated plasmin. PMID- 14734648 TI - Endoglin regulates nitric oxide-dependent vasodilatation. AB - Endoglin is a membrane glycoprotein that plays an important role in cardiovascular development and angiogenesis. We examined the role of endoglin in the control of vascular tone by measuring nitric oxide (NO)-dependent vasodilation in haploinsufficient mice (Eng+/-) and their Eng+/+ littermates. The vasodilatory effect of acetylcholine, bradykinin, and sodium nitroprusside was assessed in anesthetized mice; in isolated, perfused hindlimbs; and in aortic rings. The substantial hypotensive and vasodilatory response induced by acetylcholine and bradykinin in Eng+/+ was markedly reduced in Eng+/- mice. Both kinds of animals had similar responses to sodium nitroprusside, suggesting that the deficient vasodilatory effect is not due to a NO response impairment. Urinary and plasma concentrations of nitrites, a NO metabolite, were lower in Eng+/- than in Eng+/+ mice. The levels of endothelial nitric oxide synthase (eNOS) in kidneys and femoral arteries were about half in Eng+/- than in Eng+/+ mice and were also reduced in primary cultures of aortic endothelial cells from Eng+/- compared with those from Eng+/+ mice. Furthermore, overexpression or suppression of endoglin in cultured cells induced a marked increase or decrease in the protein levels of eNOS, respectively. Thus, our results in vivo and in vitro demonstrate a relationship between endoglin and NO-dependent vasodilation mediated by the regulation of eNOS expression. PMID- 14734649 TI - Differences in endosomal targeting of human (beta)1- and (beta)2-adrenergic receptors following clathrin-mediated endocytosis. AB - The beta(2)-adrenergic receptor (beta(2)AR) undergoes agonist-mediated endocytosis via clathrin-coated pits by a process dependent on both arrestins and dynamin. Internalization of some G protein-coupled receptors, however, is independent of arrestins and/or dynamin and through other membrane microdomains such as caveolae or lipid rafts. The human beta(1)AR is less susceptible to agonist-mediated internalization than the beta(2)-subtype, and its endocytic route, which is unknown, may be different. We have found that (i) co-expression of arrestin-2 or -3 enhanced the internalization of both subtypes whereas co expression of dominant-negative mutants of arrestin-2 or dynamin impaired their internalization, as did inhibitors of clathrin-mediated endocytosis. (ii) Agonist stimulation increased the phosphorylation of beta(2)AR but not beta(1)AR. (iii) In response to agonist, each subtype redistributed from the cell surface to a distinct population of cytoplasmic vesicles; those containing beta(1)AR were smaller and closer to the plasma membrane whereas those containing beta(2)AR were larger and more perinuclear. (iv) When subcellular fractions from agonist-treated cells were separated by sucrose density gradient centrifugation, all of the internalized beta(2)AR appeared in the lighter endosomal-containing fractions whereas some of the internalized beta(1)AR remained in the denser plasma membrane containing fractions. (v) Both subtypes recycled with similar kinetics back to the cell surface upon removal of agonist; however, recycling of beta(2)AR but not beta(1)AR was inhibited by monensin. Based on these results, we propose that the internalization of beta(1)AR is both arrestin- and dynamin-dependent and follows the same clathrin-mediated endocytic pathway as beta(2)AR. But during or after endocytosis, beta(1)AR and beta(2)AR are sorted into different endosomal compartments. PMID- 14734650 TI - The Arf activator Gea2p and the P-type ATPase Drs2p interact at the Golgi in Saccharomyces cerevisiae. AB - Arf GTPases regulate both the morphological and protein sorting events that are essential for membrane trafficking. Guanine nucleotide exchange factors (GEFs) specific for Arf proteins determine when and where Arf GTPases will be activated in cells. The yeast Gea2p Arf GEF is a member of an evolutionarily conserved family of high molecular mass Arf GEFs that are peripherally associated with membranes. Nothing is known about how these proteins are localized to membranes, and few direct binding partners have been identified. In yeast, Gea2p has been implicated in trafficking through the Golgi apparatus and in maintaining Golgi structure. A major function of the Golgi apparatus is the packaging of cargo into secretory granules or vesicles. This process occurs through a series of membrane transformation events starting with fenestration of a saccular membrane, and subsequent remodeling of the fenestrated membrane into a mesh-like tubular network. Concentration of secretory cargo into nodes of the tubular network leads to enlargement of the nodes, which correspond to forming vesicles/granules, and thinning of the surrounding tubules. The tubules eventually break to release the secretory vesicles/granules into the cytoplasm. This process is highly conserved at the morphological level from yeast to mammalian cells. Drs2p, a multi-span transmembrane domain protein and putative aminophospholipid translocase, is required for the formation of a class of secretory granules/vesicles in yeast. Here we show that Drs2p interacts directly with Gea2p, both in vitro and in vivo. We mapped the domain of interaction of Drs2p to a 20-amino-acid region of the C terminal cytoplasmic tail of the protein, adjacent to a region essential for Drs2p function. Mutations in Gea2p that abolish interaction with Drs2p are clustered in the C-terminal third of the Sec7 domain, and are important for Gea2p function. We characterize one such mutant that has a thermosensitive phenotype, and show that it has morphological defects along the secretory pathway in the formation of secretory granules/vesicles. PMID- 14734651 TI - Phosphorylation of golgin-160 by mixed lineage kinase 3. AB - Golgin-160 is a member of the coiled-coil family of golgin proteins, which are proposed to regulate the structure of the Golgi complex. The C-terminal two thirds of golgin-160 is predicted to form a coiled-coil domain and the N-terminal head domain contains several putative binding domains, regulatory motifs and phosphorylation sites. Recently, it has been demonstrated that caspase-dependent cleavage of the golgin-160 head domain occurs rapidly after induction of apoptosis. The role of golgin-160 phosphorylation and the functional implications for Golgi structure have not been defined. In this study, we investigated the kinase(s) responsible for phosphorylation of golgin-160. Signaling through the small G-protein Rac and mixed-lineage-kinase-3 (MLK3) resulted in increased phosphorylation of golgin-160. The intracellular distribution of MLK3 overlapped with that of golgin-160 and the two proteins could be co-immunoprecipitated. In vitro kinase assays demonstrated that MLK3 directly phosphorylates golgin-160 in the N-terminal head region between residues 96 and 259. Overexpression of MLK3 caused an enhanced caspase-dependent cleavage of golgin-160 at Asp139. Golgin-160 is the first non-kinase substrate of MLK3 identified, and phosphorylation by MLK3 might modulate cleavage of golgin-160 during apoptosis. PMID- 14734652 TI - The Q267E mutation in the sodium/iodide symporter (NIS) causes congenital iodide transport defect (ITD) by decreasing the NIS turnover number. AB - The Na(+)/I(-) symporter (NIS) is a key plasma membrane glycoprotein that mediates active iodide (I(-)) transport in the thyroid and other tissues. Since isolation of the cDNA encoding NIS (G. Dai, O. Levy, and N. Carrasco (1996) Nature 379, 458-460), ten mutations in NIS have been identified as causes of congenital iodide transport defect (ITD). Two of these mutations (T354P and G395R) have been thoroughly characterized at the molecular level. Both mutant NIS proteins are inactive but normally expressed and correctly targeted to the plasma membrane. The hydroxyl group at the beta-carbon of residue 354 is essential for NIS function, whereas the presence of a charged or large side-chain at position 395 interferes with NIS function. We report the extensive molecular analysis of the Q267E mutation in COS-7 cells transfected with rat or human Q267E NIS cDNA constructs. We used site-directed mutagenesis to engineer various residue substitutions into position 267. In contrast to previous suggestions that Q267E NIS was inactive, possibly because of a trafficking defect, we conclusively show that Q267E NIS is modestly active and properly targeted to the plasma membrane. Q267E NIS exhibited lower V(max) values for I(-) than wild-type NIS, suggesting that the decreased level of activity of Q267E NIS is due to a lower catalytic rate. That Q267E NIS retains even partial activity sets this ITD-causing mutant apart from T354P and G395R NIS. The presence of charged residues (of any polarity) other than Glu at position 267 rendered NIS inactive without affecting its expression or targeting, but substitution with neutral residues at this position was compatible with partial activity. PMID- 14734654 TI - Minimal mutations are required to effect a radical change in function in CEA family members of the Ig superfamily. AB - GPI anchorage in the CEA family results in the acquisition of radically different functions relative to TM anchorage, including inhibition of differentiation and anoikis, disruption of tissue architecture and promotion of tumorigenicity. CEA GPI anchors, as determined by the carboxy-terminal exon of CEA, demonstrate biological specificity in their ability to confer these functional changes. CEA family GPI anchorage appears to have evolved twice independently during the primate radiation, in a manner suggestive of evolution from more primitive TM anchored CEACAM1. We show here that very few mutations in the TM exon of present day human CEACAM1 are required to give efficient GPI anchorage and the biological specificity of CEA GPI anchors, i.e., to give the differentiation-blocking function of GPI-anchored CEA. Such a change in anchorage could therefore represent a relatively facile means for producing radical change in molecular function of Ig superfamily members during evolution. PMID- 14734653 TI - Regulation of blood-testis barrier dynamics: an in vivo study. AB - An in vivo model was used to investigate the regulation of tight junction (TJ) dynamics in the testis when adult rats were treated with CdCl(2). It was shown that the CdCl(2)-induced disruption of the blood-testis barrier (BTB) associated with a transient induction in testicular TGF-beta2 and TGF-beta3 (but not TGF beta1) and the phosphorylated p38 mitogen activated protein (MAP) kinase, concomitant with a loss of occludin and zonula occludens-1 (ZO-1) from the BTB site in the seminiferous epithelium. These results suggest that BTB dynamics in vivo are regulated by TGF-beta2/-beta3 via the p38 MAP kinase pathway. Indeed, SB202190, a specific p38 MAP kinase inhibitor, blocked the CdCl(2)-induced occludin and ZO-1 loss from the BTB. This result clearly illustrates that CdCl(2) mediates its BTB disruptive effects via the TGF-beta3/p38 MAP kinase signaling pathway. Besides, this CdCl(2)-induced occludin and ZO-1 loss from the BTB also associated with a significant loss of the cadherin/catenin and the nectin/afadin protein complexes at the site of cell-cell actin-based adherens junctions (AJs). An induction of alpha(2)-macroglobulin (a non-specific protease inhibitor) was also observed during BTB damage and when the seminiferous epithelium was being depleted of germ cells. These data illustrate that a primary disruption of the BTB can lead to a secondary loss of cell adhesion function at the site of AJs, concomitant with an induction in protease inhibitor, which apparently is used to protect the epithelium from unwanted proteolysis. alpha(2)-Macroglobulin was also shown to associate physically with TGF-beta3, afadin and nectin 3, but not occludin, E-cadherin or N-cadherin, indicating its possible role in junction restructuring in vivo. Additionally, the use of SB202190 to block the TGF-beta3/p 38 MAP kinase pathway also prevented the CdCl(2)-induced loss of cadherin/catenin and nectin/afadin protein complexes from the AJ sites, yet it had no apparent effect on alpha(2)-macroglobulin. These results demonstrate for the first time that the TGF-beta3/p38 MAP kinase signaling pathway is being used to regulate both TJ and AJ dynamics in the testis, mediated by the effects of TGF-beta3 on TJ and AJ-integral membrane proteins and adaptors, but not protease inhibitors. PMID- 14734655 TI - Loss of basement membrane, receptor and cytoskeletal lattices in a laminin deficient muscular dystrophy. AB - Basement membrane laminins bearing the alpha2-subunit interact with alpha dystroglycan and beta1-integrins, cell-surface receptors that are found within the rectilinear costameric lattices of skeletal muscle sarcolemma. Mutations of the alpha2 subunit are a major cause of congenital muscular dystrophy. To determine whether the costameres are altered as a result of laminin alpha2 mutations, the skeletal muscle surface of a dystrophic mouse (dy(2J)/dy(2J)) lacking the alpha2-LN domain was examined by confocal and widefield deconvolution immunomicroscopy. Although the dy(2J) dystrophic fibers possessed a normal appearing distribution of alpha2-laminins and alpha-dystroglycan within a rectilinear costameric lattice at 6.5 weeks of age, by 11 weeks the surface architecture of these components were found to be disorganized, with frequent effacement of the circumferential and longitudinal lattice striations. The defect in the lattice organization was also noted to be a characteristic of type IV collagen, nidogen, perlecan, beta1(D)-integrin, dystrophin and vinculin. The development of this pattern change occurring only after birth suggests that although alpha2-laminins are not essential for the initial assembly of the costameric framework, they play a role in maintaining the stability and organization of the framework. PMID- 14734656 TI - The C-terminal domain of Drosophila (beta) heavy-spectrin exhibits autonomous membrane association and modulates membrane area. AB - Current models of cell polarity invoke asymmetric cues that reorganize the secretory apparatus to induce polarized protein delivery. An important step in this process is the stabilization of the protein composition in each polarized membrane domain. The spectrin-based membrane skeleton is thought to contribute to such stabilization by increasing the half-life of many proteins at the cell surface. Genetic evidence is consistent with a negative role for Drosophila beta(Heavy)-spectrin in endocytosis, but the inhibitory mechanism has not been elucidated. Here, we investigated the membrane binding properties of the C terminal nonrepetitive domain of beta(Heavy)-spectrin through its in vivo expression in transgenic flies. We found that this region is a membrane association domain that requires a pleckstrin homology domain for full activity, and we showed for the first time that robust membrane binding by such a C terminal domain requires additional contributions outside the pleckstrin homology. In addition, we showed that expression of the beta(Heavy)-spectrin C terminal domain has a potent effect on epithelial morphogenesis. This effect is associated with its ability to induce an expansion in plasma membrane surface area. The membrane expansions adopt a very specific bi-membrane structure that sequesters both the C-terminal domain and the endocytic protein dynamin. Our data provide supporting evidence for the inhibition of endocytosis by beta(Heavy) spectrin, and suggest that the C-terminal domain mediates this effect through interaction with the endocytic machinery. Spectrin may be an active partner in the stabilization of polarized membrane domains. PMID- 14734657 TI - Role of microtubules and tea1p in establishment and maintenance of fission yeast cell polarity. AB - Microtubules and the protein tea1p have important roles in regulating cell polarity in the fission yeast Schizosaccharomyces pombe. Here, using combinations of drugs, environmental perturbations and genetic mutants, we demonstrate that once a cell polarity axis is established, microtubules have at best a minor role in maintaining the cortical actin cytoskeleton and the rate and direction of cell growth. In addition, we find that after perturbations that disrupt cell polarity and the cortical actin cytoskeleton, microtubules are not required for re establishment of polarity per se. However, after such perturbations, the distribution of cytoplasmic microtubules plays an important role in dictating the position of sites of polarity re-establishment. Furthermore, this influence of microtubule distribution on site selection during polarity re-establishment requires the presence of tea1p, suggesting that tea1p is crucial for coupling microtubule distribution to the regulation of cell polarity. Our results suggest a model in which, at the cellular level, two distinct and separable mechanisms contribute to how tea1p regulates site selection during polarity re establishment. First, tea1p remaining at cell tips after cortical depolarization can serve as a cortical landmark for microtubule-independent site selection; second, tea1p newly targeted to the cell cortex by association with microtubules can promote the formation of polarity axes de novo. PMID- 14734658 TI - Dense-core granules: a specific hallmark of the neuronal/neurosecretory cell phenotype. AB - Expression of dense-core granules, a typical exocytic organelle, is widely believed to be controlled by coordinate gene expression mechanisms specific to neurones and neurosecretory cells. Recent studies in PC12 cells, however, have suggested the number of granules/cells depends on the levels of only one of their cargo proteins, chromogranin A, regulating the metabolism of the other proteins, and thus the composition of the organelles, by an on/off switch mechanism. In addition, transfection of chromogranin A was reported to induce appearance of dense-core granules in the non-neurosecretory fibroblasts of the CV-1 line. Here the role of chromogranin A has been reinvestigated using not the heterogeneous PC12 line but several clones isolated therefrom. In these clones, investigated as such or after transfection with chromogranin A antisense sequences, the ratio between chromogranin A and its secretory protein mate, chromogranin B, was not constant but highly and apparently randomly variable. Variability of the chromogranin A/chromogranin B ratio was seen by confocal immunofluorescence also among the cells of single clones and subclones and among the granules of single cells. Moreover, stable and transient transfections of chromogranin A in a PC12 clone characterised by a low number of dense-core granules (one fifth of the reference clone) failed to modify significantly the number of the organelles, despite the several-fold increase of the granin. Finally, in three types of non neurosecretory cells (CV-1, adenocarcinoma TS/A and a clone of PC12 incompetent for secretion) the transfected chromogranin A accumulated mostly in the Golgi/transGolgi area and was released rapidly from resting cells (constitutive secretion) as revealed by both immunofluorescence during cycloheximide treatment and pulse-chase experiments. Only a minor fraction was sorted to discrete organelles that were not dense-core granules, but primarily lysosomes because they contained no chromogranin B, and were largely positive for the late endosomal-lysosomal markers, lamp1 and lamp3. Dense-core granules are therefore true hallmarks of neurones and neurosecretory cells. Their number/cell appears independent of chromogranin A and their composition does not appear to be constant; in particular, they exhibit considerable, and so far unexplained variability in the chromogranin A/chromogranin B ratio. PMID- 14734659 TI - In vivo prediction of response to antiestrogen treatment in estrogen receptor positive breast cancer. AB - In metastatic breast cancer, the estrogen receptor (ER) is a well-known prognostic factor predictive of response to hormonal treatment in most, but not all, patients. Recently, a receptor-specific radioligand for in vivo imaging of the ER in breast cancer patients was developed: (123)I-labeled cis-11beta-methoxy 17alpha-iodovinyl-estradiol (Z-(123)I-MIVE). It showed high sensitivity and specificity for the in vivo detection of ER-positive breast cancer. The aim of this study was to determine whether Z-(123)I-MIVE scintigraphy is able to predict response or resistance to antiestrogen therapy in patients with metastatic ER positive breast carcinoma. METHODS: Twenty-three patients with first metastases of their breast cancer and positive Z-(123)I-MIVE scintigraphy were included and treated with tamoxifen, 40 mg/d. Scintigraphy was repeated, on average, 4 wk later. The results of these scintigraphies were compared with the clinical outcome. RESULTS: On baseline scintigraphy, 21 of 23 patients had clear uptake and 2 of 23 patients had faint uptake of Z-(123)I-MIVE. After initiation of antiestrogen treatment, 17 of 21 patients with clear uptake on baseline scintigraphy showed complete blockade of ER activity on the Z-(123)I-MIVE scintigraphy. Four of 21 patients showed mixed or no ER blockade. All patients with faint baseline uptake or mixed or no ER blockade after tamoxifen showed progressive disease despite antiestrogen treatment. Patients with clear baseline uptake and complete ER blockade after tamoxifen had a significantly longer progression-free interval (mean +/- SEM, 14.4 +/- 1.6 vs. 1.8 +/- 0.8 mo; P < 0.01). CONCLUSION: Z-(123)I-MIVE scintigraphy seems to be a useful tool to predict response or resistance to antiestrogen treatment in ER-positive metastatic breast cancer patients and to depict nonresponders before the clinical manifestation of progression. PMID- 14734660 TI - Somatostatin receptors in malignant lymphomas: targets for radiotherapy? AB - Somatostatin (SS) receptor (sst) scintigraphy is widely used in the visualization of neuroendocrine tumors expressing sst, and radiotherapy using radionuclide labeled SS analogs has been introduced for treatment of patients with neuroendocrine tumors. Previous sst scintigraphy studies revealed that malignant lymphomas can also be visualized using this technique. The question has been addressed whether lymphomas might also be possible targets for radiotherapy using radionuclide-labeled SS analogs. Therefore, we investigated in vitro the characteristics of lymphoma tissues and lymphoid cell lines to evaluate whether lymphomas can be targets for radiotherapy. METHODS: Six orbital lymphomas, 2 Hodgkin's lymphomas, and 2 non-Hodgkin's lymphomas from the neck region were collected. Reverse transcriptase polymerase chain reaction (RT-PCR) and quantitative RT-PCR were performed to detect and quantify the expression of sst(1 5) mRNA. Receptor autoradiography studies using [(125)I-Tyr(3)]octreotide were performed to evaluate binding to sst on cryostat sections of lymphomas. Immunohistochemistry was used to investigate expression of sst(2) and sst(3). Membrane binding studies and in vitro internalization experiments using [(125)I Tyr(3)]octreotide were performed to study binding and uptake of [(125)I Tyr(3)]octreotide by lymphoid cell lines (JY, TMM, APD) and primary cells derived from a B-cell-derived chronic lymphatic leukemia. RESULTS: A selective expression of sst(2) and sst(3) messenger RNA (mRNA) was demonstrated. By quantitative RT PCR, expression levels of sst(2) and sst(3) mRNA were relatively low. Autoradiography studies revealed low binding of [(125)I-Tyr(3)]octreotide, whereas immunoreactivity could not be detected for sst(2) and sst(3) by immunohistochemistry. On the lymphoid cell lines only low numbers of high affinity SS binding sites were found. In vitro, uptake of [(125)I Tyr(3)]octreotide by these cells was also very low. CONCLUSION: On the basis of our findings, we conclude that lymphomas do not appear to be candidates for radiotherapy using radionuclide-labeled SS analogs. However, lymphomas are highly radiosensitive tumors and further clinical studies should be performed to evaluate whether the low receptor density is sufficient for targeting treatment in these tumors. PMID- 14734661 TI - USP and PET radiopharmaceuticals: 1997 FDAMA puts standard-setting body at center of regulatory process. PMID- 14734662 TI - The role of 18F-FDG PET in staging and early prediction of response to therapy of recurrent gastrointestinal stromal tumors. AB - Gastrointestinal stromal tumors (GISTs) are gaining the interest of researchers because of impressive metabolic response to the targeted molecular therapeutic drug imatinib mesylate. Initial reports suggest an impressive role for (18)F-FDG PET in follow-up of therapy for these tumors. However, the role of (18)F-FDG PET versus that of CT has not been established. Therefore, we compared the roles of (18)F-FDG PET and CT in staging and evaluation of early response to imatinib mesylate therapy in recurrent or metastatic GIST. METHODS: The study included 54 patients who underwent (18)F-FDG PET and CT scans within 3 wk before initiation of imatinib mesylate therapy. Forty-nine of these patients underwent repeat scans 2 mo after therapy. The numbers of sites or organs containing lesions on (18)F FDG PET and CT scans were compared. Corresponding lesions on (18)F-FDG PET and CT scans or those confirmed to be malignant in appearance by other imaging modalities or on follow-up were considered true positives. Lesions seen on (18)F FDG PET or CT scans but not seen or confirmed to be of benign appearance with other imaging modalities or on follow-up were considered false positives. Measurements of the maximum standard uptake value (SUV) on (18)F-FDG PET scans and tumor size on CT scans were used for quantitative evaluation of early tumor response to therapy. RESULTS: A total of 122 and 114 sites and/or organs were involved on pretherapy (18)F-FDG PET and CT scans, respectively. The sensitivity and positive predictive values (PPVs) for CT were 93% and 100%; whereas these values for (18)F-FDG PET were 86% and 98%. However, the differences between these values for CT and (18)F-FDG PET were not statistically significant (P = 0.27 for sensitivity and 0.25 for PPV). This suggests comparable performance of (18)F-FDG PET and CT in staging GISTs. Repeat scans at 2 mo after therapy showed agreement between (18)F-FDG PET and CT scans in 71.4% of patients (57.1% having a good response to therapy and 14.3% lacking a response). Discrepant results between (18)F-FDG PET and CT were recorded for 28.6% of the patients. (18)F-FDG PET predicted response to therapy earlier than did CT in 22.5% of patients during a longer follow-up interval (4-16 mo), whereas CT predicted lack of response to therapy earlier than (18)F-FDG PET in 4.1%. One patient did not undergo long-term follow-up. These findings suggest that (18)F-FDG PET is superior to CT in predicting early response to therapy in recurrent or metastatic GIST patients. CONCLUSION: The performances of (18)F-FDG PET and CT are comparable in staging GISTs before initiation of imatinib mesylate therapy. However, (18)F-FDG PET is superior to CT in predicting early response to therapy. Thus, (18)F-FDG PET is a better guide for imatinib mesylate therapy. PMID- 14734663 TI - DOE announces 20-year science facility plan. PMID- 14734664 TI - 2004 PET Learning Center Dates Set; symposia series to debut. PMID- 14734665 TI - 18F-FDG uptake in squamous cell carcinoma of the cervix is correlated with glucose transporter 1 expression. AB - This prospective study investigates the relationship between glucose transporter 1 (Glut-1) expression and PET images using (18)F-FDG and its uptake and compares them with the tumor status (primary vs. recurrent or persistent), initial grade of histologic differentiation, and International Federation of Gynecologic Obstetrics (FIGO) staging for cervical cancer patients. METHODS: A dual-phase (18)F-FDG PET scan was performed on 51 participants within the 2 wk before surgery or biopsy. (18)F-FDG uptake was quantified by calculating standardized uptake values (SUVs). After (18)F-FDG PET scanning, 51 histologically proven squamous cell carcinoma specimens were examined to determine their degree of differentiation, using hematoxylin and eosin staining, and the expression of Glut 1 by an immunohistochemical stain. Twenty normal cervical and 20 cervical intraepithelial neoplasia (CIN) sets of tissue were also used to compare the results of Glut-1 expression in these tissues. The expression of Glut-1 was the product of (the intensity [with grades 0-3, defined qualitatively]) with (percentages of the lesion area that were positive). The results of Glut-1 expression were analyzed in combination with the SUVs (SUV1 was that at 40 min and SUV2 was that at 3 h), tumor status, initial cell differentiation, and FIGO staging. RESULTS: Significant overexpression of Glut-1 was noted in 48 of the 51 (94.1%) cancer specimens. None or only minimal expression of Glut-1 was observed in basal layers of normal and CIN tissues. Significant positive correlation was observed between Glut-1 expression and the SUVs in cervical cancer specimens (r = 0.74, P < 0.000 for SUV1 and r = 0.65, P < 0.000 for SUV2). In recurrent or persistent tumor, tumor size was significantly associated with both Glut-1 expression (r = 0.508, P = 0.011) and SUV1 (r = 0. 456, P = 0.025). For recurrent or persistent tumor, only SUV1 reached statistical significance when compared with lymph node metastasis (P = 0.0226). CONCLUSION: Glut-1 expression was related to (18)F-FDG uptake in cervical cancer patients. Recurrent or persistent cervical cancer tumor had significantly higher Glut-1 expression than metastatic lymph nodes. The values of SUV and the expression of Glut-1 did not correlate with the initial grade of histologic differentiation and FIGO staging. PMID- 14734668 TI - Reagents and methods for PET using bispecific antibody pretargeting and 68Ga radiolabeled bivalent hapten-peptide-chelate conjugates. AB - The aim of this work was to develop reagents and methods potentially useful in PET, using (68)Ga in a 2-step pretargeting protocol. METHODS: We prepared bispecific antibodies (bsAbs) for disease-specific targeting of carcinoembryonic antigen-positive cells and recognition of later-administered bivalent hapten peptide conjugates. The secondary antibody arm (antibody 679) recognizes a histaminyl-succinyl-glycine (HSG) structural subunit. The bsAbs were prepared as Fab' x Fab' conjugates using chemical cross-linking methods and as bispecific diabodies using recombinant DNA technologies. A HSG-bivalent hapten conjugate bearing the macrocyclic ring chelating agent 1,4,7,10-tetraazacyclododecane N,N',N",N"'-tetraacetic acid (DOTA) was designed to be readily radiolabeled with (68)Ga taken directly from a (68)Ge/(68)Ga generator system. Reagents were tested in vitro and, then, for their targeting properties in a preclinical animal model of human cancer. RESULTS: A chemically cross-linked hMN-14 x 679 F(ab')(2) and a fully humanized bispecific diabody construct (BS1.5H), expressed in Escherichia coli, were prepared for this work. We synthesized the bivalent peptide termed IMP 241 [DOTA-Phe-Lys(HSG)-D-Tyr-Lys(HSG)-NH(2)] and labeled it with (68)Ga and (67)Ga at temperatures from 45 degrees C to 100 degrees C, over times of 15 min to 1 h, establishing 15 min at 95 degrees C as a useful condition for (68)Ga labeling. When we formulated the IMP 241 bivalent hapten-peptide with ammonium acetate buffer at pH 4-5 and eluted the (68)Ga from the generator directly into the peptide solution, we achieved an almost quantitative incorporation of the (68)Ga into IMP 241, as analyzed by size-exclusion high-performance liquid chromatography, after mixing the complex with the 679 antibody. For in vivo studies we used (67)Ga-IMP 241 as a surrogate for (68)Ga-IMP 241, in view of the short, 68-min half-life of the (68)Ga nuclide. The (67)Ga-IMP 241 was successfully pretargeted to human colon tumor xenografts in athymic mice with both the chemical and the diabody bispecific proteins. High tumor-to-normal tissue ratios for (67)Ga uptake were found for all tissues at 1 to 6 h after injection of (67)Ga-IMP 241. When using the BS1.5H diabody for pretargeting, tumor-to-blood, tumor-to-liver, and tumor-to-lung ratios of (67)Ga-IMP 241 at 1 and 3 h after injection were 41:1 and 137:1, 51:1 and 106:1, and 16:1 and 46:1, respectively. CONCLUSION: The general approach described, along with the new compositions and the labeling methods we have developed, may eventually allow for use of (68)Ga-labeled specific targeting agents in a routine clinical PET application. PMID- 14734671 TI - The usefulness of neck pinhole SPECT as a complementary tool to planar scintigraphy in primary and secondary hyperparathyroidism. AB - Pinhole SPECT (P-SPECT) has proven to be a high-resolution and sensitive method in both experimental and clinical studies. In this study, we investigated whether P-SPECT combined with conventional planar scintigraphy can give additional information in hyperfunctioning parathyroid gland detection in both primary hyperparathyroidism (pHPT) and secondary hyperparathyroidism (sHPT) since planar imaging has proved partially limited, especially in sHPT. METHODS: We studied 110 consecutive patients with HPT, selecting 67 patients who underwent neck surgery and had definitive histology: 48 with pHPT and 19 with sHPT. All patients underwent planar scintigraphy, (99m)Tc-methoxyisobutylisonitrile ((99m)Tc-MIBI) double-phase scintigraphy (n = 22) or (99m)Tc-tetrofosmin subtraction scintigraphy (n = 45), using a gamma-camera with a parallel-hole collimator. P SPECT was then performed (180 degrees; matrix size, 128 x 128; zoom factor, 2; time per frame, 40 s) using a tilted detector equipped with a pinhole collimator (insert, 4.45 mm). RESULTS: In the 48 pHPT patients, 49 lesions (43 adenomas, 1 carcinoma, and 5 hyperplastic glands, including 1 intrathyroidal) were found at surgery; in the 19 sHPT patients, 51 lesions (49 hyperplastic glands, including 1 in persistens thymus, and 2 adenomas) were found. P-SPECT proved to be a highly sensitive method, identifying more lesions than planar imaging in both pHPT (97.9% vs. 87.7%) and sHPT (92.1% vs. 78.4%), significantly (P < 0.05) in the latter. P-SPECT, positive in all adenomas, increased planar sensitivity especially in small and light-weight ones, 30.8% of which missed on planar imaging, and also identified a significantly higher number of hyperplastic glands, irrespective of their size. P-SPECT improved image quality and resolution, offering a more correct lesion localization in eutopic and ectopic sites. Neither P-SPECT nor planar imaging had false-positive findings. Moreover, P-SPECT correctly predicted the status found at surgery in 97.9% of pHPT patients and in 82.3% of sHPT patients with multigland disease, whereas planar imaging correctly predicted the status in 89.6% and 58.8%, respectively. P-SPECT was the only positive procedure in 8.9% of all patients, also revealing more lesions in 6% of sHPT patients when both methods were positive. CONCLUSION: P-SPECT appears a highly sensitive, high-resolution method. We suggest its use as a preoperative complementary tool to neck planar scintigraphy, selectively in pHPT patients but extensively in sHPT patients. PMID- 14734673 TI - 99mTc-TRODAT-1 imaging of multiple system atrophy. AB - Using (99m)Tc-TRODAT-1 ((99m)Tc-[2[[2-[[[3-(4-chlorophenyl)-8-methyl-8 azabicyclo[3,2,1]-oct-2-yl]-methyl](2-mercaptoethyl) amino]ethyl]amino]ethane thiolato(3-)-N2,N2',S2,S2]oxo-[1R-(exo-exo)])) brain SPECT imaging, we measured striatal dopamine transporters (DATs) activity in multiple system atrophy (MSA) to investigate the possibility of differentiating it from Parkinson's disease (PD) and to correlate the findings with the parkinsonian (MSA-P) and cerebellar (MSA-C) subtypes of MSA. METHODS: Forty-nine patients with probable MSA (30 MSA P, 19 MSA-C), a disease control group of 36 age- and disease duration-matched patients with probable PD, and a healthy control group of 33 age-matched healthy volunteers participated in a SPECT study. The ratios of specific striatal binding to-nonspecific occipital binding, including the striatum-to-occipital ratio (S/O), putamen-to-occipital ratio (P/O), caudate nucleus-to-occipital ratio (C/O), and putamen-to-caudate nucleus ratio (P/C), were calculated. The statistical analyses of uptakes among 4 groups used ANOVA followed by Games Howell's multiple comparisons. The Spearman correlation coefficient between the motor scores of Unified Parkinson's Disease Rating Scale (UPDRS-III) and those binding ratios of the MSA-P and MSA-C groups and the PD group was also performed. RESULTS: The striatal binding was more symmetrically reduced in the MSA-P (asymmetric index, 14.2) and MSA-C (asymmetric index, 8.1) groups, in contrast to the greater asymmetric reduction in the PD group (asymmetric index, 28.6). Overall striatal binding was significantly reduced in the MSA-P (-59.8%), MSA-C ( 29.9%), and PD (-58.0%) groups with no overlap between these values and those of the control group. Like the PD group, bilateral P/O, C/O, and S/O ratio values were significantly reduced in the MSA-P and MSA-C groups. Nevertheless, the reduction of bilateral P/O and S/O ratios was more for the MSA-P group than for the MSA-C group. P/C ratios showed that the MSA-P and PD groups had similar patterns of nigral impairment, but the MSA-C group had a different pattern. No correlation between the UPDRS-III scores and (99m)Tc-TRODAT-1 bindings was found in both MSA-P and MSA-C groups; in contrast, a significant negative correlation was noted in the PD group. CONCLUSION: (99m)Tc-TRODAT-1 brain SPECT is capable of scientifically differentiating between the MSA-P and MSA-C subtypes, and MSA-P has more symmetric nigrostriatal damage than that in PD. (99m)Tc-TRODAT-1 brain SPECT imaging probably could provide important information to differentiate MSA from PD. PMID- 14734674 TI - PET in the assessment of therapy response in patients with carcinoma of the head and neck and of the esophagus. AB - In patients with carcinoma of the head and neck and of the esophagus, metabolic and functional imaging by PET with (18)F-FDG has a pivotal role in the evaluation of tumor response to therapy, specifically, in the prediction of progression-free survival and overall survival. Metabolic imaging allows the detection of biochemical changes within tumor cells as opposed to identifiable morphologic changes. Anatomic imaging modalities do not reliably differentiate between responders and nonresponders early during the course of follow-up. The correlation between histopathologic tumor response after preoperative therapy and clinical prognosis is well established for many cancers. Squamous carcinoma of the head and neck and esophageal carcinoma demonstrate avid (18)F-FDG uptake. For these cancers, (18)F-FDG PET parallels histopathologic findings in its ability to detect residual viable tumor; therefore, it is a valuable tool for the noninvasive assessment of histopathologic tumor response in advanced-stage cases after neoadjuvant therapy before surgery. Early determination of nonresponders is of prime importance, as timely therapy modification can be accomplished for patients who do not demonstrate a response to therapy. This determination is exceptionally important for head and neck and esophageal malignancies, both of which are known for their unfavorable prognosis, as early modifications in therapy regimens for nonresponders may improve patient outcome. There is now evidence that (18)F-FDG PET is a sensitive and specific method for determining therapy response and for providing important prognostic information for these cancers. Therefore, (18)F-FDG PET may change patient management and lead to improved survival for a selected group of patients with carcinoma of the head and neck and of the esophagus. PMID- 14734675 TI - Characterization of the generation of radiolabeled monodisperse albuterol particles using the spinning-top aerosol generator. AB - Inhaled radiolabeled aerosols provide invaluable information about in vivo drug deposition. Here, we report our methodology for radiolabeling and imaging monodisperse pharmacologic aerosols in order to study basic aerosol science concepts of drug delivery within the human airways. METHODS: We used a spinning top aerosol generator to produce (99m)Tc-labeled monodisperse albuterol sulfate aerosols of 1.5-, 3-, and 6- micro m mass median aerodynamic diameter. RESULTS: In vitro Andersen cascade validation data showed that technetium and albuterol were coassociated on each impactor stage for all 3 aerosols, and the radiolabeling process itself did not affect their particle size distributions. Good-quality gamma-camera scintigraphic images of lung and extrathoracic deposition were obtained within an asthmatic patient. CONCLUSION: We have successfully radiolabeled and imaged monodisperse albuterol aerosols within the human lungs. This novel technique provides an important tool to relate fundamental concepts of aerosol particle behavior, in vivo deposition, and therapeutic clinical response. PMID- 14734676 TI - Validation of QGS and 4D-MSPECT for quantification of left ventricular volumes and ejection fraction from gated 18F-FDG PET: comparison with cardiac MRI. AB - The aim of this study was to validate Quantitative Gated SPECT (QGS) and 4D MSPECT for assessing left ventricular end-diastolic and systolic volumes (EDV and ESV, respectively) and left ventricular ejection fraction (LVEF) from gated (18)F FDG PET. METHODS: Forty-four patients with severe coronary artery disease were examined with gated (18)F-FDG PET (8 gates per cardiac cycle). EDV, ESV, and LVEF were calculated from gated (18)F-FDG PET using QGS and 4D-MSPECT. Within 2 d (median), cardiovascular cine MRI (cMRI) (20 gates per cardiac cycle) was done as a reference. RESULTS: QGS failed to accurately detect myocardial borders in 1 patient; 4D-MSPECT, in 2 patients. For the remaining 42 patients, correlation between the results of gated (18)F-FDG PET and cMRI was high for EDV (R = 0.94 for QGS and 0.94 for 4D-MSPECT), ESV (R = 0.95 for QGS and 0.95 for 4D-MSPECT), and LVEF (R = 0.94 for QGS and 0.90 for 4D-MSPECT). QGS significantly (P < 0.0001) underestimated LVEF, whereas no other parameter differed significantly between gated (18)F-FDG PET and cMRI for either algorithm. CONCLUSION: Despite small systematic differences that, among other aspects, limit interchangeability, agreement between gated (18)F-FDG PET and cMRI is good across a wide range of clinically relevant volumes and LVEF values assessed by QGS and 4D-MSPECT. PMID- 14734677 TI - Transport of 99mTc-MAG3 via rat renal organic anion transporter 1. AB - Recently, complementary DNA (cDNA) encoding a p-aminohippurate (PAH) transporter designated rat organic anion transporter 1 (OAT1) was isolated. OAT1, a multispecific organic anion transporter at the basolateral membrane, is exclusively expressed in the middle segment of the proximal tubule in the rat kidney. It has been proposed that OAT1 is indirectly involved in PAH uptake via the Na(+) dicarboxylate cotransporter. In this study, in molecular biologic experiments using OAT1-expressing Xenopus laevis oocytes, we obtained evidence that (99m)Tc-mercaptoacetylglycylglycylglycine (MAG3) is transported via OAT1. METHODS: Capped OAT1 complementary RNA (cRNA) was synthesized from library plasmid cDNA linearized with BamHI using in vitro transcription. Defolliculated oocytes were injected with 10 ng of OAT1 cRNA. Two to 3 d after injection, uptake of (99m)Tc-MAG3 was measured using ND96 solution containing 18.5 kBq of (99m)Tc MAG3. Before the uptake experiments, OAT1-expressing oocytes were preincubated for 2 h with 1 mmol/L glutarate (a dicarboxylate), to generate an outwardly directed glutarate gradient. Then, after incubation for 60 min at room temperature, radioactivity of oocytes was determined. For the inhibition experiments, uptake was assessed in the absence or presence of inhibitor: 2 mmol/L of PAH, o-iodohippurate (OIH), probenecid, 3,5-diiodo-4-pyridone-N-acetate (iodopyracet), furosemide, ethacrynic acid, glucoheptonate, maleic acid, L-Tyr, or tetraethylammonium (TEA) or 0.1 mmol/L of 2,4-dinitrophenol (DNP). RESULTS: Na(+) had a significant effect on (99m)Tc-MAG3 uptake (P < 0.05). Accumulated glutarate stimulated simultaneous (99m)Tc-MAG3 uptake and glutarate excretion (P < 0.001). The following compounds significantly inhibited (99m)Tc-MAG3 uptake: PAH, 8.5% +/- 16.2% of (99m)Tc-MAG3 uptake in the absence of an inhibitor; OIH, 26.4% +/- 21.7%; probenecid, 29.1% +/- 12.4%; iodopyracet, 15.8% +/- 7.9%; furosemide, 30.5% +/- 15.7%; ethacrynic acid, 21.6% +/- 10.6%; glucoheptonate, 35.6% +/- 22.6%; and maleic acid, 60.1% +/- 18.7%. (99m)Tc-MAG3 accumulation in Xenopus laevis oocytes was not significantly inhibited by TEA, L-Tyr, or DNP. CONCLUSION: The following substances had a cis-inhibitory effect on (99m)Tc-MAG3 transport: PAH, OIH, probenecid, iodopyracet, furosemide, ethacrynic acid, and glucoheptonate. Glutarate had a trans-stimulative effect on (99m)Tc-MAG3 transport. (99m)Tc-MAG3 acts as a substrate of OAT1, an organic anion/dicarboxylate exchanger. PMID- 14734678 TI - Transport of 99mTc-MAG3 via rat renal organic anion. PMID- 14734679 TI - Imaging of experimental colitis with a radiolabeled leukotriene B4 antagonist. AB - The use of radiolabeled leukocytes is considered the gold standard for scintigraphic imaging of inflammatory bowel disease. The disadvantages of (99m)Tc hexamethylpropyleneamine oxime (HMPAO)-leukocytes, however, encourage the search for new imaging agents with at least similar diagnostic accuracy but without the laborious preparation and subsequent risk of contamination. In this study we investigated the imaging characteristics of a new imaging agent that specifically binds to the leukotriene B(4) (LTB(4)) receptors expressed on neutrophils. Imaging characteristics of the (111)In-labeled LTB(4) antagonist (DPC11870) were compared with those of (18)F-FDG and (99m)Tc-HMPAO-granulocytes in a rabbit model of experimental colitis. METHODS: Acute colitis was induced in New Zealand White (NZW) rabbits by infusion of trinitrobenzene sulfonic acid in the descending colon. Forty-eight hours after induction of colitis, all animals were injected intravenously with (99m)Tc-granulocytes, (18)F-FDG, or (111)In-DPC11870. The pharmacokinetics and biodistribution were studied by serial scintigraphic imaging and by ex vivo counting of dissected tissues. RESULTS: All 3 radiopharmaceuticals showed the inflamed colon as early as 1 h after injection. However, compared with (99m)Tc-granulocytes, both (111)In-DPC11870 and (18)F-FDG were superior in revealing the inflamed lesions. The biodistribution data showed that uptake of (111)In-DPC11870 in the inflamed colon was highest (0.72 +/- 0.18 percentage injected dose per gram [%ID/g]), followed by uptake of (99m)Tc-granulocytes (0.40 +/- 0.11 %ID/g) and of (18)F-FDG (0.16 +/- 0.04 %ID/g). Because of low activity concentrations in the noninflamed colon, the radiolabeled LTB(4) antagonist also revealed the highest ratio of affected colon to unaffected colon (11.6 for (111)In-DPC11870, 5.5 for (99m)Tc-granulocytes, and 4.1 for (18)F-FDG). CONCLUSION: The radiolabeled LTB(4) antagonist DPC11870 clearly delineated acute colitis lesions in NZW rabbits within 1 h after injection. Because of high uptake in the inflamed lesions and a low activity concentration in the noninflamed colon, images acquired with (111)In-DPC11870 were better than those acquired with (99m)Tc-granulocytes or (18)F-FDG. PMID- 14734680 TI - In vivo investigation of estrogen regulation of adrenal and renal angiotensin (AT1) receptor expression by PET. AB - The renin angiotensin system (RAS) has been implicated as one mediator of the cardiovascular effects of estrogen. Since changes in angiotensin type 1 (AT(1)) receptor expression are central to modulation of the RAS, we used the noninvasive PET imaging technique to study for the in vivo effects of estrogen on membrane and intracellular AT(1) receptors. METHODS: Dynamic PET measurements of canine AT(1) (cAT(1)) receptors using the radiolabeled AT(1) receptor antagonist, (11)C L-159,884, were performed during 2-wk consecutive periods of estrogen deprivation induced by ovariectomy and 17beta-estradiol (E(2)) replacement. RESULTS: Kinetic modeling of time-activity curves in the kidney and adrenal showed lower receptor expression in the estrogen replete state (21% and 30% decrease in Gjedde-Patlak slope, influx constant, respectively). These in vivo findings correlated with in vitro radioligand-binding assays with (125)I-[Sar(1),Ile(8)]angiotensin II showing reduced AT(1) receptor number in the adrenal (35%), glomeruli (30%), myocardium (35%), and liver (21%) in the estrogen-replenished compared with estrogen-depleted animals. CONCLUSION: Although other endogenous systems are known to regulate AT(1) receptors and could compete with estrogenic actions, these PET studies reveal that estrogen attenuates AT(1) receptor expression in vivo. Thus, estrogen modulation of AT(1) receptors may contribute to the cardiovascular protective effects associated with estrogen. PMID- 14734681 TI - Analysis of the regional uptake of radiolabeled deoxyglucose analogs in human tumor xenografts. AB - It has been shown in vitro that the cell uptake of (18)F-FDG, a tracer of glucose metabolism, increases under hypoxia. This is consistent with increased glycolytic metabolism. We have previously shown that in ischemic heart ex vivo the rates of uptake of (18)F-FDG and 2-(14)C-deoxy-D-glucose ((14)C-2DG) are both reduced. In this study, we investigated this effect in tumors by comparing the microdistribution of (18)F-FDG and (14)C-2DG in hypoxic and normoxic regions. METHODS: Mice (MF1) bearing LS174T human tumor xenografts were injected with premixed (18)F-FDG (100 MBq), (14)C-2DG (0.37 MBq), and pimonidazole hydrochloride (60 mg/kg). After 30, 60, and 120 min, tissues (n = 4) were taken and counted for whole-body biodistribution. Tumors were frozen, sectioned, and exposed to phosphor image plates to obtain a quantitative digital image of radionuclide distribution. Sections were then stained to reveal tumor pathophysiology: Hematoxylin and eosin staining demonstrated viable and necrotic regions, and immunohistochemical staining detected pimonidazole metabolism in hypoxic cells. The images of radionuclide microdistribution and histology were then coregistered and analyzed to assess radionuclide trapping throughout the tumor on a pixel-by-pixel basis. The Pearson correlation coefficients between the 2 radionuclides were calculated. The relative amounts of nuclide were then analyzed in viable and necrotic regions and in normoxic and hypoxic regions. RESULTS: Whole-body biodistributions for the 2 radiotracers were similar. A high Pearson correlation coefficient was obtained for the 2 radionuclides throughout the tumors (r = 0.85 +/- 0.10, P < 0.0001), indicating a highly similar microdistribution. When the tumors were divided into viable and necrotic regions, the ratio of mean counts per pixel was 1.96 (P < 0.0001), whereas for hypoxic versus normoxic regions it was 1.26 (P < 0.0001). There was no significant difference in selectivity for hypoxia between the 2 radiotracers (P = 0.86). CONCLUSION: The tumor microdistribution of deoxyglucose in viable, hypoxic, and necrotic regions show that there was little change in the microdistribution of deoxyglucose throughout this time course. This study extends previous in vitro work and confirms the selectivity of deoxyglucose for viable cells over necrotic regions and for hypoxic cells over normoxic regions in vivo. PMID- 14734682 TI - Imaging taxane-induced tumor apoptosis using PEGylated, 111In-labeled annexin V. AB - 99mTc-Labeled annexin V has been used for the imaging of tumor apoptosis induced by chemotherapy. However, owing to the short half-life of annexin V, multiple injections of the radiotracer are necessary to capture the peak apoptotic activity. In this study, we evaluated the imaging properties of an (111)In labeled, long-circulating annexin V. METHODS: Both polyethylene glycol (PEG) and the metal chelator diethylenetriaminepentaacetic acid (DTPA) were simultaneously introduced to annexin V or ovalbumin through the use of a heterofunctional PEG precursor. Imaging studies were performed in mice bearing subcutaneously inoculated human mammary MDA-MB-468 tumors. The mice were treated with poly(L glutamic acid)-paclitaxel, monoclonal antibody C225, or a combination of poly(L glutamic acid)-paclitaxel and C225, followed by intravenous injection of (111)In DTPA-PEG-annexin V. Images were acquired 48 h after the injection of the radiotracer. Autoradiography and TUNEL (terminal deoxynucleotidyltransferase mediated dUTP nick-end labeling) staining were performed on adjacent tumor slices for the localization of apoptotic cells. The imaging properties of unPEGylated annexin V and PEGylated ovalbumin were also determined to permit assessment of the specificity of (111)In-DTPA-PEG-annexin V. RESULTS: Tumor apoptotic index increased from 1.67% +/- 0.31% at baseline to 7.60% +/- 0.72% and 11.07% +/- 1.81%, respectively, 4 d after treatment with poly(L-glutamic acid)-paclitaxel or combined poly(L-glutamic acid)-paclitaxel and C225. Tumor uptake (percentage of injected dose per gram of tumor [%ID/g]) of PEGylated (111)In-DTPA-PEG-annexin 4 d after treatment was significantly higher in tumors treated with poly(L-glutamic acid)-paclitaxel (10.76 +/- 1.38 %ID/g; P = 0.001) and with combined poly(L glutamic acid)-paclitaxel and C225 (9.84 +/- 2.51 %ID/g; P = 0.029) than in nontreated tumors (6.14 +/- 0.67 %ID/g), resulting in enhanced visualization of treated tumors. (111)In-DTPA-PEG-annexin V distributed into the central zone of tumors, whereas (111)In-DTPA-annexin V was largely confined to the tumor periphery. Furthermore, uptake of (111)In-DTPA-PEG-annexin V by tumors correlated with apoptotic index (r = 0.87, P = 0.02). Increase in tumor uptake of the nonspecific PEGylated protein (111)In-DTPA-PEG-ovalbumin was also observed after poly(L-glutamic acid)-paclitaxel treatment (55.6%), although this increase was less than that observed for (111)In-DTPA-PEG-annexin V (96.7%). CONCLUSION: Increased uptake of and improved visualization with (111)In-DTPA-PEG-annexin V in solid tumors after chemotherapy are mediated through both specific binding to apoptotic cells and nonspecific retention of macromolecular contrast agents in the tumors. (111)In-Labeled, PEGylated annexin V may be used to assess tumor response to chemotherapy. PMID- 14734683 TI - A gallium-labeled DOTA-alpha-melanocyte- stimulating hormone analog for PET imaging of melanoma metastases. AB - Although (18)F-FDG PET is widely used for metastatic melanoma diagnosis, it is less accurate than desirable, particularly for small foci. Since both melanotic and amelanotic melanomas overexpress receptors for alpha-melanocyte-stimulating hormone (alpha-MSH; receptor name, melanocortin type 1 receptor [MC1R]), radiolabeled alpha-MSH analogs are potential candidates for melanoma diagnosis. The aim of this study was to develop a positron emitter-labeled alpha-MSH analog suitable for PET imaging of melanoma metastases. METHODS: A short linear alpha MSH analog, [Nle(4),Asp(5),D-Phe(7)]-alpha-MSH(4-11) (NAPamide), was newly designed and conjugated to the metal chelator DOTA (1,4,7,10 tetraazacyclododecane-1,4,7,10-tetraacetic acid) to enable radiometal incorporation. Compared with our previously reported DOTA-alpha-MSH analog, DOTA MSH(oct) ([DOTA-betaAla(3),Nle(4),Asp(5),D-Phe(7),Lys(10)]-alpha-MSH(3-10)), the major modification lies in the conjugation of DOTA to the C-terminal end of the peptide via the epsilon-amino group of Lys(11), as opposed to the N-terminal alpha-amino group. After labeling with (111)In, (67)Ga, and the short-lived positron emitter (68)Ga, DOTA-NAPamide was characterized in vitro and in vivo using the mouse melanoma B16F1cell line. RESULTS: DOTA-NAPamide exhibited an almost 7-fold higher MC1R binding potency as compared with DOTA-MSH(oct). In B16F1 melanoma-bearing mice, both (111)In-DOTA-NAPamide and (67)Ga-DOTA-NAPamide behaved more favorably than (111)In-DOTA-MSH(oct). Both radiopeptides exhibited higher tumor and lower kidney uptake leading to tumor-to-kidney ratios of the 4- to 48-h area under the curve that were 4.6 times ((111)In) and 7.5 times ((67)Ga) greater than that obtained with (111)In-DOTA-MSH(oct). In addition, the 4-h kidney uptake of (67)Ga-DOTA-NAPamide could be reduced by 64% by coinjection of 15 mg L-lysine, without affecting tumor uptake. Skin primary melanoma as well as lung and liver melanoma metastases could be easily visualized on tissue section autoradiographs after systemic injection of (67)Ga-DOTA-NAPamide. The melanoma selectivity of DOTA-NAPamide was confirmed by PET imaging studies using (68)Ga DOTA-NAPamide. Tumor uptake was found to be highest when the smallest amount of peptide was administered. CONCLUSION: DOTA-NAPamide labeled with either (111)In or (67)Ga/(68)Ga is in every way superior to (111)In-DOTA-MSH(oct) in murine models of primary and metastatic melanoma, which makes it a promising agent for melanoma targeting. High-contrast images obtained in PET studies with an experimental tumor model 1 h after injection augurs well for its clinical potential as an imaging tool. PMID- 14734684 TI - Augmented 18F-FDG uptake in activated monocytes occurs during the priming process and involves tyrosine kinases and protein kinase C. AB - Activated monocytes with a high (18)F-FDG accumulation can affect the results of clinical PET studies. To better understand the mechanisms regulating monocytic (18)F-FDG uptake, we investigated the effect of priming and respiratory-burst generation and further evaluated the role of potential protein kinase pathways. METHODS: Purified human monocytes were primed with interferon-gamma (IFN-gamma), and respiratory burst was generated by stimulation of primed cells with phorbol 12-myristate-13-acetate (PMA). Oxygen-intermediate generation was assessed by luminescence measurements after the addition of lucigenin. (18)F-FDG uptake after 30 min of incubation was measured for unprimed control cells, primed cells, and PMA-stimulated cells. The role of protein kinases was investigated using respective inhibitors. RESULTS: PMA stimulation of primed monocytes dramatically increased oxygen-intermediate generation, leading to a 42.2 +/- 1.1 fold higher level of cumulative luminescence compared with unprimed control cells, whereas IFN-gamma priming alone resulted in low luminescence levels (13.9% +/- 4.6% of PMA-stimulated cells). In contrast, priming alone was sufficient to augment monocytic (18)F-FDG uptake to 273.3% +/- 16.7% of control levels (P < 0.001), and it was not further increased by PMA stimulation. The tyrosine kinase inhibitor, genistein, and the specific protein kinase C inhibitor, staurosporine, completely abolished the priming-induced enhancement of (18)F-FDG uptake and lowered uptake to control levels. Under the same conditions, wortmannin, a phosphatidylinositol 3 kinase (PI3 kinase)-specific inhibitor, and cycloheximide, a protein synthesis inhibitor, were associated with only minor reductions in the enhanced-uptake effect of priming. CONCLUSION: IFN-gamma priming alone, without stimulation of respiratory-burst activity, is sufficient to induce maximal augmentation of (18)F FDG uptake in monocytes. Furthermore, this metabolic effect appears to involve tyrosine kinases and the protein kinase C pathway but is independent of the PI3 kinase pathway. PMID- 14734685 TI - Pharmacokinetics, dosimetry, and toxicity of the targetable atomic generator, 225Ac-HuM195, in nonhuman primates. AB - Short-lived alpha-emitting isotopes individually conjugated to monoclonal antibodies have now reached human use, but little is still known about their toxicity. Use of antibody targetable (225)Ac nanogenerators is a new approach in the field of alpha-immunotherapy offering the advantage of a 10-d half-life (t(1/2)) and increased potency due to generation of 3 new atoms, yielding a total of 4 alpha-particles. However, the 3 alpha-emitting daughter elements generated have the potential for significant toxicity as these nuclides are no longer bound to the carrier IgG. METHODS: Cynomolgus monkeys were used to evaluate the toxicity of prototype (225)Ac nanogenerators. Monoclonal antibody HuM195 (anti CD33) is the carrier for planned human clinical trials of (225)Ac; there are no CD33 sites in cynomolgus monkeys. In one experiment, 2 monkeys received a single intravenous dose of (225)Ac-HuM195 at 28 kBq/kg. This dose level is approximately the planned initial human dose. In another experiment, 2 animals received a dose escalation schedule of 3 increasing (225)Ac-HuM195 doses with a cumulative activity of 377 kBq/kg. The whole-blood t(1/2) of (225)Ac, ratios of (225)Ac to its ultimate alpha-emitting daughter nuclide (213)Bi, generation of monkey anti HuM195 antibodies (MAHA), hematologic indices, serum biochemistries, and clinical parameters were measured. Monkeys were euthanized and examined histopathologically when the dose escalation reached toxicity. RESULTS: The blood t(1/2) of (225)Ac-HuM195 was 12 d, and 45% of generated (213)Bi daughters were cleared from the blood. MAHA production was not detected. Approximately 28 kBq/kg of (225)Ac caused no toxicity at 6 mo, whereas a cumulative dose of approximately 377 kBq/kg caused severe toxicity. In the cumulative dosing schedule, single doses of approximately 37 kBq/kg resulted in no toxicity at 6 wk. After approximately 130 kBq/kg were administered, no toxicity was observed for 13 wk. However, 28 wk after this second dose administration, mild anemia and increases of blood urea nitrogen and creatinine were detected. After administration of an additional 185 kBq/kg, toxicity became clinically apparent. Monkeys were euthanized 13 and 19 wk after the third dose administration (cumulative dose was 377 kBq/kg). Histopathologic evaluation revealed mainly renal tubular damage associated with interstitial fibrosis. CONCLUSION: (225)Ac nanogenerators may result in renal toxicity and anemia at high doses. The longer blood t(1/2) and the lack of target cell antigens in cynomolgus monkeys may increase toxicity compared with human application. Therefore, a dose level of at least 28 kBq/kg may be a safe starting dose in humans. Hematologic and renal function will require close surveillance during clinical trials. PMID- 14734686 TI - Uptake, localization, and dosimetry of 111in and 201tl in human testes. AB - This study concerns the testicular uptake and dosimetry of Auger electron emitting radionuclides that are used during routine diagnostic nuclear medicine procedures. To consider the possible effects of these radionuclides on spermatogenic cells, a study has been undertaken to obtain in vivo data for quantification of (111)In chloride and (201)Tl chloride uptake into the human testis after intravenous administration. Values have been determined for uptake into the testis as a whole and to the seminiferous tubules where the germ cells are located. METHODS: Data were obtained from patients with prostate cancer who opted for orchidectomy to effect hormone suppression. Patients were administered intravenously 1.5 MBq of either (111)In chloride or (201)Tl chloride at 24 or 48 h before orchidectomy. Upon removal, the testes were analyzed to assess uptake of radionuclide. Conventional dosimetry has been used to estimate testicular radiation doses using our values of percentage uptake. RESULTS: Uptake of both (111)In chloride and (201)Tl chloride into the testes was seen at a level above that explained by simple homogeneous distribution of the radionuclide throughout the body; the testes as a whole demonstrated increased uptake by factors of 3.56 and 4.01 compared with nonspecific uptake for (111)In and (201)Tl, respectively, at 24 h after administration. Both radionuclides gained access to the seminiferous tubules. CONCLUSION: The results obtained indicate that the values of testicular radiation doses quoted by the International Commission on Radiological Protection for (111)In might be too low by a factor of 4, whereas those for (201)Tl might be too high by a factor of 4. No data were obtained for uptake by individual germ cells within the testis and, therefore, no consideration of dosimetry at the cellular level was possible. However, it has been demonstrated that uptake of diagnostic Auger electron-emitting radionuclides by male germ cells within the testis is possible after intravenous administration. PMID- 14734687 TI - Noninvasive measurement of gastric accommodation by means of pertechnetate SPECT: limiting radiation dose without losing image quality. AB - The gastric accommodation response to a meal is impaired in conditions such as functional dyspepsia. At present, a barostat study is the gold standard to assess fundic relaxation in response to a meal. However, this method is invasive and possibly induces artifacts as a result of positive intraluminal balloon pressure. A noninvasive scintigraphic test has been developed to measure gastric accommodation in humans. The aim of this study was to refine this method, increasing the imaging time span and limiting the radiation dose applied without losing image quality, so that repeated measurements within 1 subject are possible without increasing radiation risk. METHODS: Thirteen healthy volunteers without gastrointestinal symptoms were recruited from a student population. Each volunteer had previously undergone a barostat study. After an overnight fast, volunteers were scanned twice on separate days after intravenous injection of 200 MBq (99m)Tc-pertechnetate. On 1 occasion, volunteers were pretreated with a proton pump inhibitor. Thirty minutes after injection, sequential, 7-min SPECT scans (72 views, 10 s/view, 128 matrix) were acquired on a dual-head gamma-camera system before and up to 2 h after ingestion of a test meal. After reconstruction (filtered backprojection, ramp-Butterworth filter; order, 10; cutoff, 0.45 Nyquist), fundus volume was calculated semiautomatically by means of a threshold voxel volume tool. RESULTS: Limiting injection dose from 370-740 MBq to 200 MBq (99m)Tc-pertechnetate resulted in good-quality images, with high target-to background ratio up to 150 min after injection. This represents a significant dose reduction, from 4.6-9.3 to 2.5 mSv. There was no significant difference between SPECT fundic volumes or accommodation response with or without proton pump inhibitor pretreatment. Volume kinetics were similar to those with barostat studies, but gastric volumes were inferior. CONCLUSION: Refining the methodology yields an improved noninvasive test for the assessment of gastric accommodation without unnecessarily increasing radiation burden. This technique enables repeated and serial measurement of gastric accommodation to a test meal, a process that is potentially useful for characterization and follow-up of dyspeptic patients with and without drug intervention. PMID- 14734688 TI - Ahem, blame the daughter! PMID- 14734689 TI - Cystic fibrosis adult care: consensus conference report. PMID- 14734690 TI - Smoking reduction and tobacco-related cancers: the more things change, the more they stay the same. PMID- 14734691 TI - Postoperative radiotherapy for breast cancer: growing evidence for an impact on survival. PMID- 14734692 TI - Improved Paclitaxel formulation hints at new chemotherapy approach. PMID- 14734693 TI - Are intellectual property rights hampering cancer research? PMID- 14734694 TI - Narrowing the field: identifying ways to improve SNP association studies. PMID- 14734696 TI - Stat bite: Relative survival rates of ovarian cancer by stage, 1992-1999. PMID- 14734697 TI - For ovarian cancer, an optimal treatment remains to be found. PMID- 14734699 TI - Tobacco and cancer: recent epidemiological evidence. PMID- 14734700 TI - Effects of reduced cigarette smoking on the uptake of a tobacco-specific lung carcinogen. AB - BACKGROUND: Limited data are available on carcinogen uptake in smokers who reduce their smoking. To determine whether reducing the number of cigarettes smoked per day would lead to a corresponding reduction in carcinogen uptake, we measured levels of metabolites of the tobacco-specific lung carcinogen 4 (methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) in the urine of smokers who reduced their smoking for up to 26 weeks. METHODS: We recruited 153 smokers, of whom 151 were randomly assigned to a reduction group or a waitlist group. In the reduction group of 102 smokers, we measured the metabolites 4-(methylnitrosamino) 1-(3-pyridyl)-1-butanol (NNAL) and its glucuronides (NNAL-Gluc) at two baseline times and at weeks 4, 6, 8, 12, and 26 after baseline. Smokers were then expected to reduce their number of cigarettes per day by 25% in weeks 0-2, 50% in weeks 2 4, and 75% in weeks 4-6 and to maintain the reduced level through week 26. In the waitlist group of 49 smokers, four baseline measurements over 7 weeks were made to assess the longitudinal stability of the metabolite measurements, and then the smokers began the reduction program. All statistical tests were two-sided. RESULTS: For waitlist and reduction groups results were comparable. Statistically significant reductions in the lung carcinogen metabolites were observed at most intervals as smokers reduced the number of cigarettes smoked each day. However, the observed decreases were generally modest, always proportionally less than the reductions in cigarettes smoked per day, and sometimes transient. For example, among the 65 individuals in the reduction group who reduced cigarettes per day by 40% or more during weeks 4-12 after baseline, mean decreases in cigarettes per day were 53% (week 4), 74% (week 6), 75% (week 8), and 74% (week 12); whereas the corresponding mean reductions in NNAL plus NNAL-Gluc were 29%, 33%, 37%, and 29%. (P<.001 for all NNAL plus NNAL-Gluc values) CONCLUSIONS: Statistically significant reductions in levels of urinary metabolites of a tobacco-specific lung carcinogen were achieved by reduction in smoking, but for most smokers, reductions were modest and transient. PMID- 14734701 TI - Breast-conserving surgery with or without radiotherapy: pooled-analysis for risks of ipsilateral breast tumor recurrence and mortality. AB - BACKGROUND: The objective of the study was to investigate whether radiotherapy or its omission after breast-conserving surgery has measurable consequences on local tumor growth and patient survival. METHODS: We conducted a pooled analysis of published randomized clinical trials that compared radiotherapy versus no radiotherapy after breast-conserving surgery. The outcomes studied were ipsilateral breast tumor recurrence and patient death from any cause. The pooled relative risks (RRs) were estimated with a random-effects model. Heterogeneity was assessed using the Cochran Q test. RESULTS: A search of the literature identified 15 trials with a pooled total of 9422 patients available for analysis. The relative risk of ipsilateral breast tumor recurrence after breast-conserving surgery, comparing patients treated with no radiotherapy or radiotherapy, was 3.00 (95% confidence interval [CI] = 2.65 to 3.40). Mortality data were available for 13 trials with a pooled total of 8206 patients. The relative risk of mortality was 1.086 (95% CI = 1.003 to 1.175), corresponding to an estimated 8.6% (95% CI = 0.3% to 17.5%) relative excess mortality if radiotherapy was omitted. CONCLUSION: Omission of radiotherapy is associated with a large increase in risk of ipsilateral breast tumor recurrence and with a small increase in the risk of patient mortality. PMID- 14734702 TI - Psychological impact of genetic counseling for familial cancer: a systematic review and meta-analysis. AB - BACKGROUND: Identification of a genetic basis underlying certain types of cancer has led to an increase in demand for genetic counseling about individual risks of the disease. We conducted a systematic review of the literature to determine the quality and strength of evidence relating to psychological outcomes of genetic counseling for familial cancer. METHODS: Six electronic databases were searched to identify controlled trials and prospective studies that examined the effect of genetic counseling on risk perception, knowledge, anxiety, cancer-specific worry, depression, and cancer surveillance. Twenty-one studies from 25 papers met inclusion criteria, including five controlled trials and 16 prospective studies. Analysis of each outcome was stratified by short-term (< or =1 month) and long term (> or = 3 months) follow-up. Trial evidence was assessed with standardized differences of the means at follow-up between intervention and comparison groups, and these data were pooled by use of random-effects meta-analysis. RESULTS: Meta analysis of controlled trials showed that genetic counseling improved knowledge of cancer genetics (pooled short-term difference = 0.70 U, 95% confidence interval [CI] = 0.15 to 1.26 U) but did not alter the level of perceived risk (pooled short-term difference = -0.10 U, 95% CI = -0.23 to 0.04 U). Prospective studies reported improvements in the accuracy of perceived risk. No effect was observed in controlled trials on general anxiety (pooled long-term effect = 0.05 U, 95% CI = -0.21 to 0.31 U) or cancer-specific worry (pooled long-term difference = -0.14 U, 95% CI = -0.35 to 0.06 U), although several prospective studies demonstrated short-term reductions in these outcomes. Few studies examined cancer surveillance behaviors, and no studies attempted to measure informed choice. CONCLUSIONS: Genetic counseling for familial cancer is associated with improvement in knowledge but does not have an adverse effect on affective outcomes. We urge further investigation of these findings through well designed, well-reported, randomized controlled trials with suitable comparison groups and additional outcome measures. PMID- 14734704 TI - Fecundity and twinning rates as measures of fertility before diagnosis of germ cell testicular cancer. AB - Previous studies have suggested an association between subfertility and testicular cancer by using fecundity and semen characteristics to measure fertility. The occurrence of twinning in offspring may be used to investigate male reproductive health, because dizygotic twinning is reduced by male subfertility. We therefore assessed number of children and offspring twinning rates among 4592 Swedish patients with testicular cancer and 12 254 control subjects. Before diagnosis, case patients had a decreased number of children (for testicular cancer, odds ratio [OR] = 0.71, 95% confidence interval [CI] = 0.62 to 0.81; at least three children compared with no children), with a lower frequency of dizygotic twinning (for unlike-sex twins, OR for the father having testicular cancer = 0.49, 95% CI = 0.22 to 1.08). The ratio of unlike-sex to same-sex twins was 0.22 among children of case patients and 0.66 among children of control subjects (adjusted P =.03, two-sided Wald test). We also found an increased occurrence of twinning after diagnosis, probably attributable to treatment for iatrogenic subfertility. Our study strongly supports evidence of an association between subfertility and the subsequent risk for testicular cancer. PMID- 14734703 TI - Effect of the methylenetetrahydrofolate reductase C677T polymorphism on chemosensitivity of colon and breast cancer cells to 5-fluorouracil and methotrexate. AB - BACKGROUND: Although single nucleotide polymorphisms may be potentially important pharmacogenetic determinants of cancer therapy, functional evidence regarding their relevance is currently lacking. The C677T polymorphism in the methylenetetrahydrofolate reductase (MTHFR) gene is associated with changes in cellular composition of folates. We hypothesized that this polymorphism may modulate the cytotoxic effect of 5-fluorouracil (5FU) and methotrexate (MTX), two commonly used chemotherapeutic agents for colon and breast cancers, because the modes of action of 5FU and MTX are critically dependent on cellular composition of folates. METHODS: Human HCT116 colon and MDA-MB-435 breast cancer cells were stably transfected with wild-type or mutant 677T human MTHFR cDNA. MTHFR enzyme activity and thermolability, intracellular folate composition, growth rate, and catalytic thymidylate synthase activity were determined. In vitro chemosensitivity to 5FU and MTX was determined using the sulforhodamine B assay. In vivo chemosensitivity of HCT116 cells to 5FU was determined in nude mice. RESULTS: Compared with cells expressing the wild-type MTHFR, HCT116 and MDA-MB 435 cells expressing the mutant 677T MTHFR had decreased MTHFR activity, MTHFR thermolability, changed intracellular folate distribution, accelerated cellular growth rate, and increased thymidylate synthase activity. The MTHFR 677T mutation increased chemosensitivity of colon and breast cancers to 5FU, but decreased chemosensitivity of breast cancer cells to MTX. In nude mice, xenografts expressing the mutant 677T MTHFR grew faster, but were more sensitive to 5FU, than xenografts expressing the wild-type protein. CONCLUSIONS: Our data provide evidence that the MTHFR C677T polymorphism affects the concentration and intracellular distribution of folates and changes the growth and chemosensitivity of colon and breast cancer cells. The MTHFR C677T polymorphism may be a useful pharmacogenetic determinant for providing rational and effective tailored chemotherapy. PMID- 14734705 TI - Retention of enrollees following a cancer diagnosis within health maintenance organizations in the Cancer Research Network. AB - Population laboratories with complete clinical information on episodes of care are needed to support research on the quality of care delivered to cancer patients. Data resources within the Cancer Research Network (CRN) may overcome many of the limitations of existing cancer databases, but their potential clinical value depends on the stability of the enrolled population. To assess this issue, we studied the retention rates among survivors of the 132 580 patients diagnosed with cancer from January 1, 1993, through December 31, 1998, who were enrolled at five health maintenance organization sites participating in the CRN. Enrollees were followed from cancer diagnosis through death, disenrollment, or the end of follow-up (i.e., December 31, 1999). The retention rate among survivors for all cancers combined at 1 and 5 years after cancer diagnosis was 96.0% (95% confidence interval [CI] = 95.9% to 96.1%) and 83.9% (95% CI = 83.4% to 84.3%), respectively. The proportion of enrollees diagnosed with cancer who remained enrolled and available for evaluation suggests that the CRN is well-suited for studies of the quality of care for cancer patients, survivorship, and long-term outcomes. PMID- 14734707 TI - NK cell compartments and their activation by dendritic cells. PMID- 14734706 TI - Circulating DNA microsatellites: molecular determinants of response to biochemotherapy in patients with metastatic melanoma. AB - Although biochemotherapy appears to be a promising treatment for metastatic melanoma, its impact remains unpredictable. Microsatellite markers for loss of heterozygosity (LOH) appear to have prognostic significance when identified in primary tumors and serum and/or plasma from cancer patients. However, their association with response to systemic therapy has yet to be assessed. To determine whether microsatellite markers are associated with response to therapy, serum from 41 patients with metastatic melanoma, drawn before the initiation of biochemotherapy, was analyzed for LOH with nine microsatellite markers. During a median follow-up of 13 months, the overall response rate for these 41 patients was 56%, including 13 (32%) complete responses and 10 (24%) partial responses. LOH was detected in sera from 12 (29%) of the 41 patients. The response rate of these 12 patients was 17% (95% confidence interval [CI] = 5% to 45%), whereas that of the 29 patients without LOH was 72% (95% CI = 54% to 85%) (P =.001). All statistical tests were two-sided. The presence of LOH was statistically significant and independently associated with disease progression (multivariable analysis, P =.003). Circulating tumor DNA markers may be useful in assessing prognosis for advanced melanoma patients and their response to biochemotherapy. PMID- 14734708 TI - Cutting edge: distinct TCR- and CD28-derived signals regulate CD95L, Bcl-xL, and the survival of primary T cells. AB - TCR-driven clonal expansion of T cells is limited by activation-induced cell death through CD95/CD95L interactions. This cell-contact dependent mechanism is attenuated by costimulation through CD28. Here, we show that primary rat lymph node T cells activated by "superagonistic" CD28-specific mAb, which do not require TCR-engagement for full T cell activation, do not up-regulate CD95L. CD28 superagonist activated T cells are highly resistant to artificial CD95 cross linking, and display a marked up-regulation of the survival factor Bcl-x(L). Consistently, NF-kappaB factors, known to promote Bcl-x(L) transcription, are strongly activated by superagonistic CD28 mAb stimulation. In contrast, a weaker induction of NFAT, which positively regulates the CD95L gene, in CD28 activated cells as compared with TCR- or TCR/CD28-stimulated cells was observed. Thus, by recruiting the mitogenic activity of CD28 in the absence of TCR engagement, the anti-apoptotic signals provided by costimulation are revealed without interfering proapoptotic effects induced by TCR stimulation. PMID- 14734709 TI - Induction of tumor regression by administration of B7-Ig fusion proteins: mediation by type 2 CD8+ T cells and dependence on IL-4 production. AB - CD28 signals contribute to either type 1 or type 2 T cell differentiation. Here, we show that administration of B7.2-Ig fusion proteins to tumor-bearing mice induces tumor regression by promoting the differentiation of antitumor type 2 CD8(+) effector T cells along with IL-4 production. B7.2-Ig-mediated regression was not induced in IL-4(-/-) and STAT6(-/-) mice. However, it was elicited in IFN gamma(-/-) and STAT4(-/-) mice. By contrast, IL-12-induced tumor regression occurred in IL-4(-/-) and STAT6(-/-) mice, but not in IFN-gamma(-/-) and STAT4(-/ ) mice. Moreover, B7.2-Ig treatment was effective in a tumor model not responsive to IL-12. B7.2-Ig administration elicited elevated levels of IL-4 production. Tumor regression was predominantly mediated by CD8(+) T cells, although the induction of these effector cells required CD4(+) T cells. Tumor regression induced by CD8(+) T cells alone was inhibited by neutralizing the IL-4 produced during B7.2-Ig treatment. Thus, these results indicate that stimulation in vivo of CD28 with B7.2-Ig in tumor-bearing mice results in enhanced induction of antitumor type 2 CD8(+) T cells (Tc2) leading to Tc2-mediated tumor regression. PMID- 14734711 TI - Tolerance induction by intrathymic expression of P0. AB - Genetic deficiency or instability of myelin protein zero (P0) results in hereditary motor sensory neuropathy. In view of recent advances in gene therapy, substitution of the molecular defect may become realistic in the near future. Here we investigate the impact of genetic deficiency of P0 on selection of the autoreactive T cell repertoire in the corresponding mouse model. We show that P0 mRNA transcripts are expressed in thymic stroma, similar to other myelin proteins and that expression of intact P0 protein can be detected by Western blot. Using a library of overlapping 20mer peptides spanning the entire length of P0 and applying the ELISPOT technique, we detected a strong immune response toward P0 extracellular domain peptide aa 41-60 in P0(-/-) knockout mice, but not in heterozygous P0(+/-) or wild-type (wt) mice. In addition, one cryptic epitope and two subdominant epitopes of P0 were identified. Using P0(-/-) into wt bone marrow (BM) chimeras we found that P0 expression in the host suffices for full tolerance induction, which is in line with its presence in thymic stroma. However, repopulation of P0(-/-) mice with wt BM led to partial induction of tolerance, suggesting that BM derived cells can also express this protein. Our findings may have implications for secondary autoimmunity developing after gene therapy in hereditary neuropathies and other diseases with genetically determined protein deficiency, because the repaired protein will then represent a foreign, nontolerized Ag. PMID- 14734710 TI - Mechanistic insights into impaired dendritic cell function by rapamycin: inhibition of Jak2/Stat4 signaling pathway. AB - The suppressive effect of rapamycin on T cells has been extensively studied, but its influence on the function of APC is less clear. The data in this study demonstrated that immunostimulatory activity of B10 (H2(b)) dendritic cells (DC) exposed to rapamycin (rapa-DC) was markedly suppressed as evidenced by the induction of low proliferative responses and specific CTL activity in allogeneic (C3H, H2(k)) T cells. Administration of rapa-DC significantly prolonged survival of B10 cardiac allografts in C3H recipients. Treatment with rapamycin did not affect DC expression of MHC class II and costimulatory molecules or IL-12 production. Rapamycin did not inhibit DC NF-kappaB pathway, however, IL-12 signaling through Janus kinase 2/Stat4 activation was markedly suppressed. Indeed, Stat4(-/-) DC similarly displayed poor allostimulatory activity. The Stat4 downstream product, IFN-gamma, was also inhibited by rapamycin, but DC dysfunction could not solely be attributed to low IFN-gamma production as DC deficient in IFN-gamma still exhibited vigorous allostimulatory activity. Rapamycin did not affect DC IL-12R expression, but markedly suppressed IL 18Ralpha and beta expression, which may in turn down-regulate DC IL-12 autocrine activation. PMID- 14734712 TI - Cellular maturation defects in Bruton's tyrosine kinase-deficient immature B cells are amplified by premature B cell receptor expression and reduced by receptor editing. AB - In the mouse, Bruton's tyrosine kinase (Btk) is essential for efficient developmental progression of CD43(+)CD2(-) large cycling into CD43(-)CD2(+) small resting pre-B cells in the bone marrow and of IgM(high) transitional type 2 B cells into IgM(low) mature B cells in the spleen. In this study, we show that the impaired induction of cell surface changes in Btk-deficient pre-B cells was still noticeable in kappa(+) immature B cells, but was largely corrected in lambda(+) immature B cells. As lambda gene rearrangements are programmed to follow kappa rearrangements and lambda expression is associated with receptor editing, we hypothesized that the transit time through the pre-B cell compartment or receptor editing may affect the extent of the cellular maturation defects in Btk-deficient B cells. To address this issue, we used 3-83 mu delta transgenic mice, which prematurely express a complete B cell receptor and therefore manifest accelerated B cell development. In Btk-deficient 3-83 mu delta mice, the IgM(+) B cells in the bone marrow exhibited a very immature phenotype (pre-BCR(+)CD43(+)CD2(-)) and were arrested at the transitional type 1 B cell stage upon arrival in the spleen. However, these cellular maturation defects were largely restored when Btk deficient 3-83 mu delta B cells were on a centrally deleting background and therefore targeted for receptor editing. Providing an extended time window for developing B cells by enforced expression of the antiapoptotic gene Bcl-2 did not alter the Btk dependence of their cellular maturation. We conclude that premature B cell receptor expression amplifies the cellular maturation defects in Btk deficient B cells, while extensive receptor editing reduces these defects. PMID- 14734713 TI - Tc1 and Tc2 effector cell therapy elicit long-term tumor immunity by contrasting mechanisms that result in complementary endogenous type 1 antitumor responses. AB - Cytolytic CD8(+) effector cells fall into two subpopulations based on cytokine secretion. Type 1 CD8(+) T cells (Tc1) secrete IFN-gamma, whereas type 2 CD8(+) T cells (Tc2) secrete IL-4 and IL-5. Both effector cell subpopulations display predominantly perforin-dependent cytolysis in vitro. Using an OVA-transfected B16 lung metastases model, we show that adoptively transferred OVA-specific Tc1 and Tc2 cells induce considerable suppression, but not cure, of pulmonary metastases. However, long-term tumor immunity prolonged survival times indefinitely and was evident by resistance to lethal tumor rechallenge. At early stages after therapy, protection by Tc2 and Tc1 effector cells were dependent in part on effector cell derived IL-4, IL-5, and IFN-gamma, respectively. Whereas effector cell-derived perforin was not necessary. Over time the numbers of both donor cells diminished to low, yet still detectable, levels. Concomitantly, Tc1 and Tc2 effector cell therapies potentiated endogenous recipient-derived antitumor responses by inducing 1) local T cell-derived chemokines associated with type 1-like immune responses; 2) elevated levels of recipient-derived OVA tetramer-positive CD8 memory T cells that were CD44(high), CD122(+), and Ly6C(high) that predominantly produced IFN-gamma and TNF-alpha; and 3) heightened numbers of activated recipient-derived Th1 and Tc1 T cell subpopulations expressing CD25(+), CD69(+), and CD95(+) cell surface activation markers. Moreover, both Tc2 and Tc1 effector cell therapies were dependent in part on recipient-derived IFN-gamma and TNF alpha for long-term survival and protection. Collectively, Tc1 and Tc2 effector cell immunotherapy mediate long-term tumor immunity by different mechanisms that subsequently potentiate endogenous recipient-derived type 1 antitumor responses. PMID- 14734714 TI - IL-11 protects human microvascular endothelium from alloinjury in vivo by induction of survivin expression. AB - IL-11 can reduce tissue injury in animal models of inflammation but the mechanism(s) is unknown. When C.B-17 SCID/beige mice bearing human skin grafts are injected i.p. with human PBMC allogeneic to the donor skin, infiltrating T cells destroy human microvessels by day 21. Intradermal injection of human IL-11 (500 ng/day) delays the time course of graft microvessel loss without reducing the extent of T cell infiltration. Protective actions of IL-11 are most pronounced on day 15. IL-11 has no effect on T cell activation marker, effector molecule, cytokine expression, or endothelial ICAM-1 expression. IL-11 up regulates the expression of survivin, a cytoprotective protein, in graft keratinocytes and endothelial cells. Topical application of survivin antisense oligonucleotide down-regulates survivin expression in both cell types and largely abrogates the protective effect of IL-11. We conclude that in this human transplant model, IL-11 exerts a cytoprotective rather than anti-inflammatory or immunomodulatory effect mediated through induction of survivin. PMID- 14734715 TI - Mature human thymocytes migrate on laminin-5 with activation of metalloproteinase 14 and cleavage of CD44. AB - We have previously shown that laminin-5 is expressed in the human thymic medulla, in which mature thymocytes are located. We now report that laminin-5 promotes migration of mature medullary thymocytes, whereas it has no effect on cortical immature thymocytes. Migration was inhibited by blocking mAbs directed against laminin-5 integrin receptors and by inhibitors of metalloproteinases. Interactions of thymocytes with laminin-5 induced a strong up-regulation of active metalloproteinase-14. However, we found that thymocytes did not cleave the laminin-5 gamma(2) chain, suggesting that they do not use the same pathway as epithelial cells to migrate on laminin-5. Interactions of thymocytes with laminin 5 also induced the release of a soluble fragment of CD44 cell surface molecule. Moreover, CD44-rich supernatants induced thymocyte migration in contrast with supernatants depleted in CD44 by immunoadsorption. CD44 cleavage was recently reported to be due to metalloproteinase-14 activation and led to increased migration in cancer cells. Thus, in this study, we show that laminin-5 promotes human mature thymocyte migration in vitro via a multimolecular mechanism involving laminin-5 integrin receptors, metalloproteinase-14 and CD44. These data suggest that, in vivo, laminin-5 may function in the migration of mature thymocytes within the medulla and be part of the thymic emigration process. PMID- 14734716 TI - CCL17 and IL-10 as effectors that enable alternatively activated macrophages to inhibit the generation of classically activated macrophages. AB - Classically activated macrophages (CAMphi) have been described as a major effector cell on the host's innate immunities. However, CAMphi are not generated in immunocompromised hosts whose alternatively activated macrophages (AAMphi) predominate. In this study, the mechanism by which AAMphi suppress the ability of resident macrophages (RMphi) to generate CAMphi was investigated. AAMphi were isolated from peritoneal exudates of mice 2 days after third-degree thermal injuries affecting 15% total body surface area. CAMphi were generated from RMphi (peritoneal Mphi from normal mice) through stimulation with CpG DNA, a typical CAMphi inducer. RMphi did not polarize to CAMphi when they were cultured with AAMphi in a dual-chamber Transwell even when supplemented with CpG DNA. In addition, RMphi stimulated with CpG DNA did not convert to CAMphi when they were cultured with the culture fluids of AAMphi (AAMphi Culture-Sup). AAMphi Culture Sup contained IL-6, IL-10, CCL17, PGE(2), and TGF-beta. Among these, CCL17 and IL 10 inhibited CAMphi generation. The ability of AAMphi Culture-Sup to inhibit CAMphi generation was eliminated when the Culture-Sup was treated with a mixture of mAbs directed against CCL17 and IL-10. These results indicate that CCL17 and IL-10 released from AAMphi inhibit CAMphi generation from RMphi stimulated with CpG DNA. PMID- 14734717 TI - Independent control of Ly49g alleles: implications for NK cell repertoire selection and tumor cell killing. AB - A novel murine NK cell-reactive mAb, AT8, was generated. AT8 recognizes Ly49G from 129/J, BALB/c, and related mouse strains, but does not bind to Ly49G(B6). Costaining with AT8 and a Ly49G(B6)-restricted Ab (Cwy-3) provides the first direct evidence that Ly49G protein is expressed from both alleles on a significant proportion of NK cells from four different types of F(1) hybrid mice. The observed level of biallelic Ly49G expression reproducibly followed the product rule in both freshly isolated and cultured NK cells. Surprisingly, the percentage of NK cells expressing both Ly49G alleles could be dramatically increased in vitro and in vivo through IL-2R- and IFN receptor-dependent signaling pathways, respectively. Unexpectedly, Ly49G(B6+) NK cells in an H-2(d), but not H-2(b), background were more likely to lyse D(d+) and Chinese hamster ovary tumor cells than Ly49G(BALB/129+) NK cells. Furthermore, Ly49G(B6+) NK cells also proliferated to a higher degree in response to poly(I:C) than NK cells expressing a non-Ly49G(B6) allele in an H-2(d), but not H-2(b), background. These results suggest that Ly49G(B6) has a lower affinity for H-2D(d) than Ly49G(BALB/129), and the genetic background calibrates the responsiveness of NK cells bearing self-specific Ly49. Other H-2D(d) receptors on the different Ly49G(+) NK cell subsets were unequally coexpressed, possibly explaining the disparate responses of Ly49G(B6+) NK cells in different hybrid mice. These data indicate that the stochastic mono- and biallelic expression of divergent Ly49G alleles increases the range of MHC affinities and the functional potential in the total NK cell population of heterozygous mice. PMID- 14734718 TI - Estrogen preferentially promotes the differentiation of CD11c+ CD11b(intermediate) dendritic cells from bone marrow precursors. AB - Sex biases in autoimmunity and infection suggest that steroid sex hormones directly modulate immune cells. We show in this study that 17-beta-estradiol (E2) promotes the differentiation of functional dendritic cells (DC) from murine bone marrow precursor cells. Remarkably, ex vivo DC differentiation was inhibited in steroid hormone-deficient medium, and was restored by addition of physiological amounts of E2, but not dihydrotestosterone. DC differentiation was inhibited by the estrogen receptor (ER) antagonists ICI 182,780 and tamoxifen, and from ERalpha(-/-) bone marrow cells, indicating that E2 acted via ERs. E2 addition was most effective in promoting DC differentiation immediately ex vivo, but did not increase DC proliferation. E2 treatment specifically promoted differentiation of a CD11c(+) CD11b(int) DC population that displayed high levels of cell surface MHC class II and CD86, suggesting that E2 could augment numbers of potent APC. DC that differentiated in E2-supplemented medium were fully functional in their capability to mediate presentation of self and foreign Ags and stimulate the proliferation of naive CD4(+) T cells. The requirement for estrogen during DC differentiation suggests a mechanism by which E2 levels in peripheral tissues might modulate both the number and functional capabilities of DC in vivo, thereby influencing immune responses. PMID- 14734719 TI - Lck is required for activation-induced T cell death after TCR ligation with partial agonists. AB - TCR engagement can induce either T cell proliferation and differentiation or activation-induced T cell death (AICD) through apoptosis. The intracellular signaling pathways that dictate such a disparate fate after TCR engagement have only been partially elucidated. Non-FcR-binding anti-CD3 mAbs induce a partial agonist TCR signaling pattern and cause AICD on Ag-activated, cycling T cells. In this study, we examined TCR signaling during the induction of AICD by anti-CD3 fos, a non-FcR-binding anti-CD3 mAb. This mAb activates Fyn, Lck, and extracellular signal-regulated kinase, and induces phosphorylation of Src-like adapter protein, despite the inability to cause calcium mobilization or TCR polarization. Anti-CD3 fos also fails to effectively activate zeta-associated protein of 70 kDa or NF-kappaB. Using Ag-specific T cells deficient for Fyn or Lck, we provide compelling evidence that activation of Lck is required for the induction of AICD. Our data indicate that a selective and distinct TCR signaling pattern is required for AICD by TCR partial agonist ligands. PMID- 14734720 TI - Chimeric donor cells play an active role in both induction and maintenance phases of transplantation tolerance induced by mixed chimerism. AB - Donor hemopoietic cell engraftment is considered to be an indicator of allograft tolerance. We depleted chimerism with cells specifically presensitized to the bone marrow donor to investigate its role in mixed chimera-induced tolerance. Three experimental models were used: model A, B10.A cells presensitized to B6 (a anti-b cells) were injected into (B6 x D2)F(1) --> B10.A mixed chimeras grafted with DBA/2 skin; model B, anti-B6 presensitized cells prepared in DBA/2 --> B10.A mixed chimeras, thus unresponsive to DBA/2 (a anti-b/tol-d cells), were injected into (B6 x D2)F(1) --> B10.A mixed chimeras grafted with DBA/2 skin; and model C, (BALB/c x B6)F(1) cells presensitized to CBA (d/b anti-k cells) were injected into (B6 x CBA)F(1) --> BALB/c mixed chimeras grafted with B6 skin. Skin was grafted on day 30. Injection of each cell type before skin grafting abolished hemopoietic cell engraftment and prevented allograft acceptance. Injection of presensitized cells after skin grafting resulted in different outcomes depending on the models. In model A, injection of a anti-b cells completely depleted chimerism and caused allograft rejection. In model B, injection of a anti-b/tol-d cells markedly reduced, but did not deplete, peripheral chimerism and maintained skin allograft survival. In model C, d/b anti-k cells reduced chimerism to the background levels but failed to cause graft rejection, probably due to persistence of injected cells which share MHC with skin grafts. Together, the results show that presence of chimeric donor cells is essential in both the induction and maintenance phases of tolerance induced by mixed chimerism. PMID- 14734721 TI - Importance of IL-10 for CTLA-4-mediated inhibition of tumor-eradicating immunity. AB - In this study, we show that engagement of CTLA-4 on tumor-infiltrating lymphocytes from low-dose melphalan (L-phenylalanine mustard (L-PAM))-treated MOPC-315 tumor bearers led to IL-10 secretion. In addition, the inhibitory activity of CTLA-4 ligation for IFN-gamma secretion following stimulation with anti-CD3 plus anti-CD28 mAb depended on IL-10 production. Consistent with the importance of IL-10 for CTLA-4-mediated inhibition, administration of neutralizing anti-IL-10 mAb to low-dose L-PAM-treated MOPC-315 tumor bearers (administration of blocking anti-CTLA-4 mAb) resulted in enhanced tumor infiltrating lymphocyte-mediated anti-MOPC-315 cytotoxicity and led to complete tumor eradication in a higher percentage of mice than that observed with low-dose L-PAM alone. The percentage of MOPC-315 tumor-bearing mice cured following administration of neutralizing anti-IL-10 mAb to low-dose L-PAM-treated MOPC-315 tumor bearers was comparable to that observed following administration of blocking anti-CTLA-4 mAb. Moreover, IL-10 neutralization together with CTLA-4 blockade did not provide added therapeutic benefits to low-dose L-PAM-treated MOPC-315 tumor bearers. Taken together, these results indicate that CTLA-4 blockade improves the therapeutic outcome of low-dose L-PAM for MOPC-315 tumor bearers by inhibiting IL-10 secretion as a consequence of blocking CTLA-4 ligation. PMID- 14734722 TI - The abundant NK cells in human secondary lymphoid tissues require activation to express killer cell Ig-like receptors and become cytolytic. AB - Natural killer cells are important cytolytic cells in innate immunity. We have characterized human NK cells of spleen, lymph nodes, and tonsils. More than 95% of peripheral blood and 85% of spleen NK cells are CD56(dim)CD16(+) and express perforin, the natural cytotoxicity receptors (NCRs) NKp30 and NKp46, as well as in part killer cell Ig-like receptors (KIRs). In contrast, NK cells in lymph nodes have mainly a CD56(bright)CD16(-) phenotype and lack perforin. In addition, they lack KIRs and all NCR expression, except low levels of NKp46. The NK cells of tonsils also lack perforin, KIRs, NKp30, and CD16, but partially express NKp44 and NKp46. Upon IL-2 stimulation, however, lymph node and tonsilar NK cells up regulate NCRs, express perforin, and acquire cytolytic activity for NK-sensitive target cells. In addition, they express CD16 and KIRs upon IL-2 activation, and therefore display a phenotype similar to peripheral blood NK cells. We hypothesize that IL-2 can mobilize the NK cells of secondary lymphoid tissues to mediate natural killing during immune responses. Because lymph nodes harbor 40% and peripheral blood only 2% of all lymphocytes in humans, this newly characterized perforin(-) NK cell compartment in lymph nodes and related tissues probably outnumbers perforin(+) NK cells. These results also suggest secondary lymphoid organs as a possible site of NK cell differentiation and self-tolerance acquisition. PMID- 14734723 TI - Production of donor T cells is critical for induction of donor-specific tolerance and maintenance of chimerism. AB - Nonmyeloablative conditioning has significantly reduced the morbidity associated with bone marrow transplantation. The donor hemopoietic cell lineage(s) responsible for the induction and maintenance of tolerance in nonmyeloablatively conditioned recipients is not defined. In the present studies we evaluated which hemopoietic stem cell-derived components are critical to the induction of tolerance in a total body irradiation-based model. Recipient B10 mice were pretreated with mAbs and transplanted with allogeneic B10.BR bone marrow after conditioning with 100-300 cGy total body irradiation. The proportion of recipients engrafting increased in a dose-dependent fashion. All chimeric recipients exhibited multilineage donor cell production. However, induction of tolerance correlated strictly with early production of donor T cells. The chimeras without donor T cells rejected donor skin grafts and demonstrated strong antidonor reactivity in vitro, while possessing high levels of donor chimerism. These animals lost chimerism within 8 mo. Differentiation into T cells was aborted at a prethymic stage in recipients that did not produce donor T cells. Moreover, donor Ag-driven clonal deletion of recipient T cells occurred only in chimeras with donor T cells. These results demonstrate that donor T cell production is critical in the induction of transplantation tolerance and the maintenance of durable chimerism. In addition, donor T cell production directly correlates with the deletion of potentially alloreactive cells. PMID- 14734725 TI - Zwitterionic polysaccharides stimulate T cells with no preferential V beta usage and promote anergy, resulting in protection against experimental abscess formation. AB - Zwitterionic polysaccharides (Zps) from pathogenic bacteria, such as Bacteroides fragilis, are virulence factors responsible for abscess formation associated with intra-abdominal sepsis. The underlying cellular mechanism for abscess formation requires T cell activation. Conversely, abscess formation can be prevented by prophylactic s.c. injection of purified Zps alone, a process also dependent on T cells. Hence, the modulatory role of T cells in abscess formation was investigated. We show that Zps interact directly with T cells with fast association/dissociation kinetics. Vbeta repertoire analysis using RT-PCR demonstrates that Zps have broad Vbeta usage. Zps-specific hybridomas responded to a variety of other Zps, but not to a nonzwitterionic polysaccharide, indicating cross-reactivity between different Zps. Furthermore, Zps-reactive T cell hybridomas could effectively transfer protection against abscess formation. Analysis of the proliferative capacity of T cells recovered from Zps-treated animals revealed that these T cells are anergic to subsequent stimulation by the different Zps or to alloantigens in an MLR. This anergic response was relieved by addition of IL-2. Taken together, the data show that this class of polysaccharides interacts directly with T cells in a nonbiased manner to elicit an IL-2-dependent anergic response that confers protection against abscess formation. PMID- 14734724 TI - Zeta-associated protein of 70 kDa (ZAP-70), but not Syk, tyrosine kinase can mediate apoptosis of T cells through the Fas/Fas ligand, caspase-8 and caspase-3 pathways. AB - The TCR zeta-chain-associated protein of 70 kDA (ZAP-70) and Syk tyrosine kinases play critical roles in regulating TCR-mediated signal transduction. They not only share some overlapped functions but also may play unique roles in regulating the function and development of T cells. However, it is not known whether they have different effects on the activation and activation-induced cell death of T cells. To address this question, we generated cDNAs encoding chimeric molecules that a tailless TCR zeta-chain was directly linked to truncated ZAP-70 (Z/ZAP) or Syk (Z/Syk) molecules lacking the two Src homology 2 domains. Transfection of these molecules into zeta-chain-deficient cells restored their TCR expression. In addition, Z/ZAP and Z/Syk transfectants but not control cells demonstrated kinase activities in phosphorylating an exogenous substrate specific for ZAP-70 and Syk kinases. Z/ZAP transfectants activated through TCRs underwent a faster time course of apoptosis and had a greater percentage of apoptotic cells than that of Z/Syk and control cells. Activated Z/ZAP transfectants increased Fas and Fas ligand (FasL) expression 3- and 40-fold, respectively. Blocking of the Fas/FasL interaction could inhibit the apoptosis of Z/ZAP transfectants. In contrast, although activated Z/Syk transfectants could increase FasL expression, their Fas expression actually decreased and the percentage of apoptotic cells did not increase. Further studies of the mechanisms revealed that activation of Z/ZAP but not Z/Syk transfectants resulted in rapid activation of caspase-3 and caspase-8 that could also be inhibited by blocking Fas/FasL interaction. These results demonstrated that ZAP-70 and Syk play distinct roles in T cell activation and activation-induced cell death. PMID- 14734726 TI - Effect of chronic viral infection on epitope selection, cytokine production, and surface phenotype of CD8 T cells and the role of IFN-gamma receptor in immune regulation. AB - Regulation of CD8 T cell responses in chronic viral infections is not well understood. In this study, we have compared the CD8 T cell responses to immunodominant and subdominant epitopes during an acute and a chronic lymphocytic choriomeningitis virus (LCMV) infection in mice. The epitope hierarchy of the primary CD8 T cell response was similar in acute and chronic LCMV infections. However, strikingly, the epitope hierarchy of the primary CD8 T cell response was conserved in the T cell memory only in an acute but not in a chronic LCMV infection. Interestingly, in an acute infection, increasing the viral dose caused significant changes in the epitope hierarchy of the LCMV-specific memory CD8 T cell pool, with no effect on the primary CD8 T cell response. Functional and phenotypic analyses revealed that exposure of CD8 T cells to extended periods of antigenic stimulation could lead to long-term defects in cytokine production and alteration in expression of cell surface L-selectin (CD62L). Whereas expression of CD44 was minimally altered, a greater proportion of LCMV-specific memory CD8 T cells were CD62L(low) in mice that have recovered from a chronic LCMV infection, compared with acutely infected mice. Mechanistic studies showed that IFN-gammaR deficiency altered the epitope hierarchy of the pool of LCMV-specific memory CD8 T cells without significantly affecting the immunodominance of the primary CD8 T cell response in an acute infection. Taken together, these findings should further our understanding about the regulation of T cell responses in human chronic viral infections. PMID- 14734727 TI - CpG-DNA aided cross-priming by cross-presenting B cells. AB - Covalent linkage of immunostimulatory CpG-DNA to OVA (CpG-OVA complex) results in CpG-DNA-aided cross-presentation of OVA by dendritic cells (DCs). In this study, we analyzed the thesis that CpG-OVA complexes may be cross-presented by B cells to route internalized Ag into the class I MHC presentation pathway. First, we describe that conjugation of CpG-DNA to OVA enhances up to 40-fold internalization of OVA by B cells, which in turn generate the CD8 T cell epitope SIINFEKL complexed to MHC class I, albeit less efficiently than DCs. Furthermore, upon internalization, CpG-DNA conjugated to OVA stimulates B cells to up-regulate costimulatory molecules and cytokines including IL-12. Adoptive transfer of CpG OVA complex-loaded wild-type B cells cross-primes naive CD8 T cells both in wild type mice and in MyD88-deficient mice. Overall, these findings disclose attributes of B cells, including cross-presentation of exogenous Ag and cross priming of naive CD8 T cells that hitherto have been considered as hallmarks restricted to DCs. PMID- 14734728 TI - Tumor-infiltrating lymphocyte secretion of IL-6 antagonizes tumor-derived TGF beta 1 and restores the lymphokine-activated killing activity. AB - IL-6 is a multifunctional cytokine that regulates cell growth, differentiation, and cell survival. Many tumor cells produce TGF-beta1, which allows them to evade CTL-mediated immune responses. IL-6 antagonizes TGF-beta1 inhibition of CD3 cell activation. However, whether IL-6 restores NK activity, which also is suppressed by TGF-beta1, is not known. We used canine transmissible venereal tumor (CTVT), which produces TGF-beta1, as a model to determine whether IL-6 restores lymphokine-activated killer (LAK) activity. During the progression phase, CTVT cells stop expressing MHC molecules. During the regression phase, the number of surface MHC molecules increases dramatically on about one-third of tumor cells. Tumor cells that stop expressing MHC should be targeted by NK cells. In this study, we found that TGF-beta1 secreted by CTVT cells suppressed LAK cytotoxicity. Interestingly, tumor-infiltrating lymphocytes (TIL) isolated from regressing CTVT secrete high concentrations of IL-6 and antagonize the anti-LAK activity of tumor cell TGF-beta1. TIL also produce IL-6 during progression phase, but the concentration is too low to block the anti-LAK activity of TGF-beta1. There is probably a threshold concentration of IL-6 needed to reverse TGF-beta1 inhibited LAK activity. In addition, in the absence of TGF-beta1, IL-6 derived from TIL does not promote the activity of LAK. This new mechanism, in which TIL manufacture high concentrations of IL-6 to block tumor TGF-beta1 anti-LAK activity, has potential applications in cancer immunotherapy and tumor prognosis. PMID- 14734729 TI - Expression of dual TCR on DO11.10 T cells allows for ovalbumin-induced oral tolerance to prevent T cell-mediated colitis directed against unrelated enteric bacterial antigens. AB - The triggering Ag for inflammatory bowel disease and animal models of colitis is not known, but may include gut flora. Feeding OVA to DO11.10 mice with OVA specific transgenic (Tg) TCR generates Ag-specific immunoregulatory CD4(+) T cells (Treg) cells. We examined the ability of oral Ag-induced Treg cells to suppress T cell-mediated colitis in mice. SCID-bg mice given DO11.10 CD4(+)CD45RB(high) T cells developed colitis, and cotransferring DO11.10 CD45RB(low)CD4(+) T cells prevented CD4(+)CD45RB(high) T cell-induced colitis in the absence of OVA. The induction and prevention of disease by DO11.10 CD4(+) T cell subsets were associated with an increase in endogenous TCRalpha chain expression on Tg T cells. Feeding OVA to SCID-bg mice reconstituted with DO11.10 CD4(+)CD45RB(high) attenuated the colitis in association with increased TGF-beta and IL-10 secretion, and decreased proliferative responses to both OVA and cecal bacteria Ag. OVA feeding also attenuated colitis in SCID-bg mice reconstituted with a mix of BALB/c and DO11.10 CD45RB(high) T cells, suggesting that OVA induced Treg cells suppressed BALB/c effector cells. The expression of endogenous non-Tg TCR allowed for DO11.10-derived T cells to respond to enteric flora Ag. Furthermore, feeding OVA-induced Treg cells prevented colitis by inducing tolerance in both OVA-reactive and non-OVA-reactive T cells and by inducing Ag nonspecific Treg cells. Such a mechanism might allow for Ag-nonspecific modulation of intestinal inflammation in inflammatory bowel disease. PMID- 14734730 TI - Early adenoviral gene expression mediates immunosuppression by transduced dendritic cell (DC): implications for immunotherapy using genetically modified DC. AB - Long-lasting, high-level gene expression in the absence of a toxic or inflammatory response to viral Ags is necessary for the successful application of genetically modified dendritic cell (DC). We previously demonstrated that efficient transduction of mature DC using DeltaE1DeltaE3 adenoviruses suppressed their stimulatory capacity for T cells. The current study was designed to investigate in more detail the suppressive effect of Ad-DC. We demonstrate that immunosuppression is not mediated by alterations in the T cell phenotype or cytokine profiles released by stimulated T cells. Also DC phenotypes are not affected. However, we demonstrate a cell cycle arrest of the T cell population stimulated by adenovirally transduced DC. Surprisingly, only freshly transduced DC are perturbed in their stimulatory capacity. Experiments using cycloheximide to block early intracellular viral gene expression showed that viral genes expressed in DC are responsible for this transient immunosuppression. In agreement with these findings, high-capacity (gutless) Ad-vectors that differ in viral gene expression from conventional DeltaE1DeltaE3 adenovirus are suitable for an efficient transduction of human DC. DC transduced with gutless Ad-vectors showed a high allostimulatory capacity for CD4(+) and CD8(+) T cells. Thus, the immunosuppressive effect of DeltaE1DeltaE3 Ad-transduced mature DC seems to be the result of early viral gene expression in DC that can be prevented using gutless Ad-vectors for transduction. These results have important implications for the use of genetically modified DC for therapeutic application. PMID- 14734731 TI - CD4+ and CD8+ regulatory T cells generated ex vivo with IL-2 and TGF-beta suppress a stimulatory graft-versus-host disease with a lupus-like syndrome. AB - Regulatory T cells generated ex vivo from conventional mouse T cells have been used to prevent and alter the course of a stimulatory graft-vs-host disease with a lupus-like syndrome. DBA/2 mouse T cells induce this syndrome when injected into (DBA/2 x C57BL/6) F(1) mice. Stimulating DBA/2 T cells with irradiated C57BL/6 in the presence of IL-2 and TGF-beta induced both CD4(+) and CD8(+) cells to develop potent suppressive activity and enhanced their survival. The IL-2 and TGF-beta-treated T cells lost their ability to induce graft-vs-host disease and, instead, prevented other parental T cells from inducing lymphoid hyperplasia, B cell activation, and an immune complex glomerulonephritis. Moreover, a single transfer of TGF-beta-conditioned T cells to animals that had already developed anti-dsDNA Abs decreased the titer, suppressed proteinuria, and doubled survival. This study raises the possibility that autologous regulatory T cells generated ex vivo have the potential to be used as an adoptive immunotherapy to induce allograft tolerance and to control autoimmunity. PMID- 14734732 TI - IL-21 induces tumor rejection by specific CTL and IFN-gamma-dependent CXC chemokines in syngeneic mice. AB - IL-21 is an immune-stimulatory four alpha helix cytokine produced by activated T cells. To study the in vivo antitumor activities of IL-21, TS/A murine mammary adenocarcinoma cells were genetically modified to secrete IL-21 (TS/A-IL-21). These cells developed small tumors that were subsequently rejected by 90% of s.c. injected syngeneic mice. Five days after injection, TS/A-IL-21 tumors showed numerous infiltrating granulocytes, NK cells, and to a lesser extent CD8(+) T cells, along with the expression of TNF-alpha, IFN-gamma, and endothelial adhesion molecules ICAM-1 and VCAM-1. At day 7, CD8(+) and CD4(+) T cells increased together with IFN-gamma, and the CXC chemokines IFN-gamma-inducible protein 10, monokine induced by IFN-gamma, and IFN-inducible T cell alpha chemoattractant. The TS/A-IL-21 tumor displayed a disrupted vascular network with abortive sprouting and signs of endothelial cell damage. In vivo depletion experiments by specific Abs showed that rejection of TS/A-IL-21 cells required CD8(+) T lymphocytes and granulocytes. When injected in IFN-gamma-deficient mice, TS/A-IL-21 cells formed tumors that regressed in only 29% of animals, indicating a role for IFN-gamma in IL-21-mediated antitumor response, but also the existence of IFN-gamma-independent effects. Most immunocompetent mice rejecting TS/A-IL-21 cells developed protective immunity against TS/A-pc (75%) and against the antigenically related C26 colon carcinoma cells (61%), as indicated by rechallenge experiments. A specific CTL response against the gp70-env protein of an endogenous murine retrovirus coexpressed by TS/A and C26 cells was detected in mice rejecting TS/A-IL-21 cells. These data suggest that IL-21 represents a suitable adjuvant in inducing specific CTL responses. PMID- 14734733 TI - Deceptive multilineage reconstitution analysis of mice transplanted with hemopoietic stem cells, and implications for assessment of stem cell numbers and lineage potentials. AB - Hemopoietic stem cells (HSC) are identified through their unique ability, at the single cell level, to long-term reconstitute all blood cell lineages. Sustained myeloid reconstitution is considered the hallmark of HSC, because myeloid progenitors and their progeny have very short half-lives. Here we demonstrate that the established practice of relying on RB6-8C5 as a myeloid specific Ab can result in overestimation of HSC frequencies because the RB6-8C5 Ab also detects Ags expressed on a sizeable population of CD3(+)CD8(+) T cells, constitutively as well as following transplantation. Likewise, a high fraction of mice transplanted with limiting numbers of ex vivo expanded Lin(-)Sca(+)kit(+)CD34(-) HSC show long term RB6-8C5(+)CD3(+) (lymphoid) but no RB6-8C5(+)CD3(-) (myeloid) reconstitution. Most noteworthy, the use of RB6-8C5 as a myeloid specific Ab can be deceptive by implicating the existence of lineage-restricted HSC capable of long-term reconstituting the myeloid and T, but not B, cell lineage. Because cross-lineage expression of "lineage-specific" markers is unlikely to be unique to the blood system, claims of unexpected cell fates should be substantiated not only by acquisition of lineage-specific markers, but also absence of markers of other lineages normally derived from the investigated stem cells. PMID- 14734734 TI - Autoreactive B cells in lupus-prone New Zealand black mice exhibit aberrant survival and proliferation in the presence of self-antigen in vivo. AB - To identify defects in B cell tolerance that may contribute to the production of autoantibodies in New Zealand Black (NZB) mice, we crossed soluble hen egg white lysozyme (sHEL) and anti-HEL Ig transgenes (Ig Tg) onto the NZB background. In this study, we have examined one of the first checkpoints involved in maintenance of peripheral B cell tolerance, follicular exclusion and elimination of self reactive B cells in the absence of T cell help. Freshly isolated anti-HEL Ig Tg B cells were labeled with CFSE, adoptively transferred into sHEL recipients, and the fate of self-reactive anti-HEL Ig Tg B cells was followed using flow cytometry and immunofluorescence microscopy. Although anti-HEL Ig Tg B cells from NZB mice are appropriately excluded from B cell follicles in NZB sHEL recipient mice, they demonstrate aberrant survival, proliferation, and generation of anti HEL Ab-producing cells. This abnormal response results from an intrinsic defect in NZB B cells, requires the presence of CD4(+) T cells, and is facilitated by the splenic environment in NZB mice. Thus, NZB mice have immune defects that interact synergistically to allow autoreactive B cells to become activated despite the presence of tolerizing autoantigens. PMID- 14734735 TI - Phosphatidylinositol 3-kinase is a determinant of responsiveness to B cell antigen receptor-mediated Epstein-Barr virus activation. AB - B cell Ag receptor (BCR) cross-linking with anti-Ig Abs efficiently induces activation of latently infected EBV in some B cell lines, but not in others. The present study was aimed at defining the molecular mechanisms that determine the response to BCR-mediated EBV activation. Comparison of Burkitt's lymphoma-derived Akata, Mutu-I, and Daudi cells, which are representative responders and nonresponders to BCR-mediated EBV activation, respectively, indicated that three signaling pathways, phosphatidylinositol 3-kinase (PI3K), extracellular signal regulated kinase (ERK), and p38 mitogen-activated protein kinase (MAPK), were activated in anti-Ig-treated Akata and Mutu-I cells. However, in anti-Ig-treated Daudi cells PI3K was not activated, ERK was faintly activated, and p38 MAPK was constitutively phosphorylated irrespective of anti-Ig treatment. Restoration of PI3K activity with insulin-like growth factor 1 restored ERK and p38 MAPK pathways, and was accompanied by EBV activation in anti-Ig-treated Daudi cells. In contrast, a specific inhibitor for PI3K, wortmannin, inhibited EBV activation by anti-Ig Abs in Akata and Mutu-I cells. Transfection assays in EBV-negative Daudi cells revealed that PI3K activated a promoter for BZLF1, which is a switch of EBV activation from a latent infection, in the absence of other EBV products suggesting that the BZLF promoter was a target of BCR signaling, and that PI3K was important for BCR-mediated BZLF1 activation. These results indicate that the absence of PI3K impedes the progression of signals through the BCR and becomes a determinant of unresponsiveness to BCR-mediated EBV activation. PMID- 14734736 TI - Intraocular tumor antigen drains specifically to submandibular lymph nodes, resulting in an abortive cytotoxic T cell reaction. AB - Ocular immune privilege is considered essential in the protection against sight threatening immune responses, as illustrated by the ability of the ocular environment to permit the growth of tumors that are rejected when implanted at other sites. Although several studies indicate that soluble Ag can drain directly into the spleen when injected into the anterior chamber, the primary site of intraocular tumor Ag presentation to tumor-specific CTLs has not been studied. To gain a better understanding of the mechanism involved in ocular immune privilege, we examined to which lymphoid organs anterior chamber tumor Ags primarily drain. Our data show that intraocular tumor Ag drains exclusively to the submandibular lymph nodes, resulting in activation of tumor-specific CTLs, whereas no Ag drainage was found in spleen. However, these tumor-specific CTLs do not distribute systemically and, as a consequence, intraocular tumor growth is unhampered. A similar lack of CTL efficacy has been observed in mice bearing s.c. tumors, which is converted to a systemic tumoricidal CTL response by administration of agonistic anti-CD40 mAb. In contrast, systemic anti-CD40 treatment of eye tumor-bearing mice did not result in mobilizing tumor-specific CTLs or tumor eradication. Together, these results show that intraocular tumor Ag drains to regional lymph nodes for activation of tumor-specific CTLs. However, the induced tumor-specific immunity is insufficient for tumor clearance, even combined with otherwise highly effective immune intervention protocols. PMID- 14734737 TI - E-selectin, thymus- and activation-regulated chemokine/CCL17, and intercellular adhesion molecule-1 are constitutively coexpressed in dermal microvessels: a foundation for a cutaneous immunosurveillance system. AB - The success of the cutaneous immune system reflects its ability to rapidly and efficiently recruit leukocytes to areas of trauma and infection. Skin-homing memory T cells expressing cutaneous lymphocyte-associated Ag tether on the walls of postcapillary venules in inflamed skin via interaction with endothelial E selectin and roll in response to the shear stress imparted by flowing blood. Rolling cells sample the vascular surface for chemoattractant compounds (e.g., thymus- and activation-regulated chemokine/CCL17 interacting with CCR4 on the leukocyte surface) and, if successfully stimulated, progress to firm arrest and transmigration mediated by LFA-1 and vascular ICAM-1. Although it is established that this sequence of events draws T cells into inflamed skin, the mechanisms directing trafficking of T cells to noninflamed skin are less well characterized. We hypothesized that basal expression and colocalization of E-selectin, chemokine (e.g., CCL17), and ICAM-1 in dermal vessels could serve to recruit T cells to noninflamed human skin. Immunohistochemical staining for E-selectin and CD31 demonstrated E-selectin expression in a restricted subset of dermal vessels in noninflamed human skin from three different sites. Confocal multicolor immunofluorescence imaging revealed a nonuniform distribution of E-selectin in dermal vessels as well as colocalization of E-selectin with CCL17 and ICAM-1. Coexpression of these molecules on blood vessels in noninflamed skin provides the basis for a model of cutaneous immunosurveillance system active in the absence of pathologic inflammation. PMID- 14734738 TI - Intravenous injection of a lentiviral vector encoding NY-ESO-1 induces an effective CTL response. AB - Lentiviral vectors can efficiently transduce a variety of nondividing cells, including APCs. We assessed the immunogenicity of a lentiviral vector encoding the melanoma Ag NY-ESO-1 in HLA-A2 transgenic mice. Direct i.v. injection of NY ESO-1 lentivirus induced NY-ESO-1(157-165)-specific CD8(+) cells, detected ex vivo with an A2/H-2K(b) chimeric class I tetramer. These NY-ESO-1(157-165) specific CD8(+) cells could be expanded by boosting with an NY-ESO-1 vaccinia virus and could kill NY-ESO-1(157-165) peptide-pulsed targets in vivo. Such direct lentiviral vector injection was similar in potency to the injection of in vitro-transduced dendritic cells (DC). In addition, human monocyte-derived DC transduced by the NY-ESO-1 lentivirus stimulated an NY-ESO-1(157-165)-specific specific CTL clone. These data suggest that direct lentiviral transduction of DC in vivo might provide a powerful immunotherapeutic strategy. PMID- 14734739 TI - IFN-gamma promotes Fas ligand- and perforin-mediated liver cell destruction by cytotoxic CD8 T cells. AB - To study liver cell damage by CTL, CD8 T cells from P14 TCR transgenic (tg) mice specific for the gp33 epitope of lymphocytic choriomeningitis virus with either deficiency in IFN-gamma (P14.IFN-gamma(null)), functional Fas ligand (P14.gld), or perforin (P14.PKO) were transferred into H8 tg mice ubiquitously expressing gp33 Ag. Treatment of H8 recipient mice with agonistic anti-CD40 Abs induced vigorous expansion of the transferred P14 T cells and led to liver cell destruction determined by increase of glutamate dehydrogenase serum levels and induction of caspase-3 in hepatocytes. Liver injury was mediated by the Fas/Fas ligand (FasL) pathway and by perforin, because P14.gld and P14.PKO T cells failed to induce increased glutamate dehydrogenase levels despite strong in vivo proliferation. In addition, H8 tg mice lacking Fas were resistant to the pathogenic effect of P14 T cells. Besides FasL and perforin, IFN-gamma was also required for liver cell damage, because P14.IFN-gamma(null) T cells adoptively transferred into H8 mice failed to induce disease. Moreover, Fas expression on hepatocytes from H8 recipient mice was increased after transfer of wild-type compared with P14.IFN-gamma(null) T cells, and wild-type P14 T cells expressed higher levels of FasL than P14 T cells lacking IFN-gamma. Thus, our data suggest that IFN-gamma released by activated CD8 T cells upon Ag contact facilitates liver cell destruction. PMID- 14734740 TI - Escherichia coli expressing recombinant antigen and listeriolysin O stimulate class I-restricted CD8+ T cells following uptake by human APC. AB - Vaccination against cancer or intracellular pathogens requires stimulation of class I-restricted CD8(+) T cells. It is therefore important to develop Ag delivery vectors that will promote cross-presentation by APCs and stimulate appropriate inflammatory responses. Toward this goal, we tested the potential of Escherichia coli as an Ag delivery vector in in vitro human culture. Bacteria expressing enhanced green fluorescent protein were internalized efficiently by dendritic cells, as shown by flow cytometry and fluorescence microscopy. Phenotypic changes in DC were observed, including up-regulation of costimulatory molecules and IL-12p40 production. We tested whether bacteria expressing recombinant Ags could stimulate human T cells using the influenza matrix protein as a model Ag. Specific responses against an immunodominant epitope were seen using IFN-gamma ELISPOT assays when the matrix protein was coexpressed with listeriolysin O, but not when expressed alone. THP-1 macrophages were also capable of stimulating T cells after uptake of bacteria, but showed slower kinetics and lower overall levels of T cell stimulation than dendritic cells. Increased phagocytosis of bacteria induced by differentiation of THP-1 increased their ability to stimulate T cells, as did opsonization. Presentation was blocked by proteasome inhibitors, but not by lysosomal protease inhibitors leupeptin and E64. These results demonstrate that recombinant E. coli can be engineered to direct Ags to the cytosol of human phagocytic APCs, and suggest possible vaccine strategies for generating CD8(+) T cell responses against pathogens or tumors. PMID- 14734741 TI - Characterization of defective CD4-CD8- T cells in murine tumors generated independent of antigen specificity. AB - Immune-based therapy confers limited benefits to hosts bearing late-stage tumors. Mounting evidence points to local suppression of T cell function as the most substantial barrier to effective antitumor immunity in hosts with large tumor burdens. Despite this, events responsible for locally defective T cells and immune suppression in tumors remain unclear. We describe in this study a predominant T cell population localized within two murine tumors that is characterized by expression of apoptotic markers and TCRalphabeta/CD3, but not CD4, CD8, or NK-associated markers. These defective cells resembled double negative (DN) T cells in lpr mice, harbored defects in the expression of T cell signaling molecules, and produced the anti-inflammatory cytokine, IL-10. Conditions known to increase or decrease the accumulation of lpr DN T cells had corresponding effects on local DN tumor infiltrating lymphocyte (TIL) levels and inversely impacted host survival. Adoptive transfer into s.c. tumors demonstrated that naive CD8(+) T cells were highly susceptible to becoming DN TIL, and local supplementation of tumors with nontumor Ag-bearing MHC class I-expressing fibroblasts decreased both this susceptibility and endogenous DN TIL levels. These findings identify a major defective T cell population with suppressive potential within tumors. The data also suggest that local T cell defectiveness is controlled by the tumor environment independent of cognate Ag specificity per se. Decreasing defective DN TIL levels by increasing noncognate peptide MHC class I availability, or modulating TCR or cytokine signaling may facilitate host survival by bolstering endogenous immunity to late-stage tumors, and may help improve therapeutic tumor vaccines. PMID- 14734742 TI - Regulation of c-Jun N-terminal kinase by MEKK-2 and mitogen-activated protein kinase kinase kinases in rheumatoid arthritis. AB - The mitogen-activated protein kinase (MAPK) c-Jun N-terminal kinase (JNK) is a critical regulator of collagenase-1 production in rheumatoid arthritis (RA). The MAPKs are regulated by upstream kinases, including MAPK kinases (MAPKKs) and MAPK kinase kinases (MAP3Ks). The present study was designed to evaluate the expression and regulation of the JNK pathway by MAP3K in arthritis. RT-PCR studies of MAP3K gene expression in RA and osteoarthritis synovial tissue demonstrated mitogen-activated protein kinase/ERK kinase kinase (MEKK) 1, MEKK2, apoptosis-signal regulating kinase-1, TGF-beta activated kinase 1 (TAK1) gene expression while only trace amounts of MEKK3, MEKK4, and MLK3 mRNA were detected. Western blot analysis demonstrated immunoreactive MEKK2, TAK1, and trace amounts of MEKK3 but not MEKK1 or apoptosis-signal regulating kinase-1. Analysis of MAP3K mRNA in cultured fibroblast-like synoviocytes (FLS) showed that all of the MAP3Ks examined were expressed. Western blot analysis of FLS demonstrated that MEKK1, MEKK2, and TAK1 were readily detectable and were subsequently the focus of functional studies. In vitro kinase assays using MEKK2 immunoprecipitates demonstrated that IL-1 increased MEKK2-mediated phosphorylation of the key MAPKKs that activate JNK (MAPK kinase (MKK)4 and MKK7). Furthermore, MEKK2 immunoprecipitates activated c-Jun in an IL-1 dependent manner and this activity was inhibited by the selective JNK inhibitor SP600125. Of interest, MEKK1 immunoprecipitates from IL-1-stimulated FLS appeared to activate c-Jun through the JNK pathway and TAK1 activation of c-Jun was dependent on JNK, ERK, and p38. These data indicate that MEKK2 is a potent activator of the JNK pathway in FLS and that signal complexes including MEKK2, MKK4, MKK7, and/or JNK are potential therapeutic targets in RA. PMID- 14734744 TI - Blockade of lymphotoxin signaling inhibits the clinical expression of murine graft-versus-host skin disease. AB - Adhesion molecules are essential for the recruitment of T cells into the skin during the development of graft-vs-host skin disease (GVHSD). However, the mechanisms responsible for the regulation of expression of cutaneous adhesion molecules in this setting are still poorly understood. In this study we blocked lymphotoxin (LT) signaling in a murine model of minor histocompatibility Ag system mismatch GVHSD by using an LTbeta receptor-Ig fusion protein (LTbetaR-Ig). The recipient mice treated with control human Ig developed clinically apparent, severe skin lesions. However, none of the mice treated with LTbetaR-Ig developed clinical skin disease. The expression of ICAM-1 in cutaneous tissue was also much lower in mice treated with LTbetaR-Ig than in mice treated with human Ig. Thus, the inhibition of LT signaling via LTbetaR-Ig treatment appears to be capable of markedly ameliorating the development of GVHSD, possibly by inhibiting the expression of adhesion molecules. PMID- 14734743 TI - The role of TNF-related activation-induced cytokine-receptor activating NF-kappa B interaction in acute allograft rejection and CD40L-independent chronic allograft rejection. AB - We analyzed the role of TNF-related activation-induced cytokine (TRANCE), a member of the TNF family expressed on activated T cells that shares functional properties with CD40L, and its receptor-activating NF-kappaB (RANK) which is mostly expressed on mature dendritic cells, during allogenic responses in vivo using a rodent heart allograft model. TRANCE mRNA was strongly up-regulated in acutely rejected allografts on days 4 and 5 posttransplantation whereas RANK was detected as early as day 1 but did not show further up-regulation during the first week. Immunofluoresence analyses of heart allografts showed that 80 and 100% of TRANCE and RANK-expressing cells were T cells and APCs, respectively. We show for the first time that short-term TRANCE blockade using a mouse RANKIg fusion molecule can significantly prolong heart allograft survival in both rat and mouse models. Similarly, rat heart allografts transduced with a RANKIg encoding recombinant adenovirus exhibited a significant prolongation of survival (14.3 vs 7.6 days, p < 0.0001). However, TRANCE blockade using RANKIg did not appear to inhibit allogeneic T and B cell priming humoral responses against RANKIg. Interestingly, TRANCE blockade induced strong up-regulation of CD40 ligand (CD40L) mRNA in allografts. Combined CD40L and TRANCE blockade resulted in significantly decreased chronic allograft rejection lesions as well as allogeneic humoral responses compared with CD40L blockade alone. We conclude that TRANCE RANK interactions play an important role during acute allograft rejection and that CD40L-independent allogeneic immune responses can be, at least in part, dependent on the TRANCE pathway of costimulation. PMID- 14734745 TI - Ontogeny of gamma delta T cells in humans. AB - T cell receptors consist either of an alpha-chain combined with a beta-chain or a gamma-chain combined with a delta-chain. alphabeta T cells constitute the majority of T cells in human blood throughout life. Flow cytometric analyses presented in this study, which focus on the representation of the developmental (naive and memory) subsets of gammadelta T cells, show by function and phenotype that this lineage contains both naive and memory cells. In addition, we show that the representation of naive T cells is higher among alphabeta than gammadelta T cells in adults and that the low frequency of naive gammadelta T cells in adults reflects ontological differences between the two major gammadelta subsets, which are distinguished by expression of Vdelta1 vs Vdelta2 delta-chains. Vdelta1 cells, which mirror alphabeta cells with respect to naive representation, predominate during fetal and early life, but represent the minority of gammadelta cells in healthy adults. In contrast, Vdelta2 cells, which constitute the majority of adult gammadelta cells, show lower frequencies of naive cells than Vdelta1 early in life and show vanishingly small naive frequencies in adults. In essence, nearly all naive Vdelta2 cells disappear from blood by 1 year of life. Importantly, even in children less than 1 year old, most of the nonnaive Vdelta2 cells stain for perforin and produce IFN-gamma after short-term in vitro stimulation. This represents the earliest immunological maturation of any lymphocyte compartment in humans and most likely indicates the importance of these cells in controlling pathology due to common environmental challenges. PMID- 14734746 TI - IFN-alpha induced adenosine production on the endothelium: a mechanism mediated by CD73 (ecto-5'-nucleotidase) up-regulation. AB - CD73 (ecto-5'-nucleotidase; EC 3.1.3.5) participates in lymphocyte binding to endothelial cells and converts extracellular AMP into a potent anti-inflammatory substance adenosine. However, the regulation of expression and function of CD73 has remained largely unknown. In this study, we show that IFN-alpha produces a time- and dose-dependent long-term up-regulation of CD73 on endothelial cells, but not on lymphocytes both at protein and RNA levels. Moreover, CD73-mediated production of adenosine is increased after IFN-alpha treatment on endothelial cells, resulting in a decrease in the permeability of these cells. Subsequent to induction with PMA, FMLP, dibutyryl cAMP, thrombin, histamine, IL-1beta, TNF alpha, and LPS, no marked changes in the level of CD73 expression on endothelial cells are observed. We also show that CD73 is up-regulated in vivo on the vasculature after intravesical treatment of urinary bladder cancers with IFN alpha. In conclusion, distinct behavior of lymphocyte and endothelial CD73 subsequent to cytokine treatment further emphasizes the existence of cell type specific mechanisms in the regulation of CD73 expression and function. Overall, these results suggest that IFN-alpha is a relevant in vivo regulator of CD73 in the endothelial-leukocyte microenvironment in infections/inflammations, and thus has a fundamental role in controlling the extent of inflammation via CD73 dependent adenosine production. PMID- 14734747 TI - Nuclear pro-IL-16 regulation of T cell proliferation: p27(KIP1)-dependent G0/G1 arrest mediated by inhibition of Skp2 transcription. AB - The precursor for IL-16 (pro-IL-16) is a nuclear and cytoplasmic PDZ domain containing protein. In this study we have found that pro-IL-16 is absent or mutated in four T lymphoblastic leukemia cell lines examined. Ectopic expression of pro-IL-16 in pro-IL-16-negative Jurkat cells blocks cell cycle progression from G(0)/G(1) to S phase associated with elevated levels of the cyclin-dependent kinase inhibitor p27(KIP1). Pro-IL-16 decreases p27(KIP1) degradation by reducing transcription and subsequent expression of Skp2, a key component of the SCF(Skp2) ubiquitin E3 ligase complex. Taken together, these findings identify pro-IL-16 as a novel regulator of Skp2 expression and p27(KIP1) levels and implicate a role for pro-IL-16 in T cell proliferation. PMID- 14734748 TI - The nonclassical MHC class I molecule Qa-1 forms unstable peptide complexes. AB - The MHC class Ib molecule Qa-1 is the primary ligand for mouse CD94/NKG2A inhibitory receptors expressed on NK cells, in addition to presenting Ags to a subpopulation of T cells. CD94/NKG2A receptors specifically recognize Qa-1 bound to the MHC class Ia leader sequence-derived peptide Qdm. Qdm is the dominant peptide loaded onto Qa-1 under physiological conditions and this peptide has an optimal sequence for binding to Qa-1. Peptide dissociation experiments demonstrated that Qdm dissociates from soluble or cell surface Qa-1(b) molecules with a t(1/2) of approximately 1.5 h at 37 degrees C. In comparison, complexes of an optimal peptide (SIINFEKL) bound to the MHC class Ia molecule H-2K(b) dissociated with a t(1/2) in the range from 11 to 31 h. In contrast to K(b), the stability of cell surface Qa-1(b) molecules was independent of bound peptides, and several observations suggested that empty cell surface Qa-1(b) molecules might be unusually stable. Consistent with the rapid dissociation rate of Qdm from Qa-1(b), cells become susceptible to lysis by CD94/NKG2A(+) NK cells under conditions in which new Qa-1(b)/Qdm complexes cannot be continuously generated at the cell surface. These results support the hypothesis that Qa-1 has been selected as a specialized MHC molecule that is unable to form highly stable peptide complexes. We propose that the CD94/NKG2A-Qa-1/Qdm recognition system has evolved as a rapid sensor of the integrity of the MHC class I biosynthesis and Ag presentation pathway. PMID- 14734749 TI - The C4A and C4B isotypic forms of human complement fragment C4b have the same intrinsic affinity for complement receptor 1 (CR1/CD35). AB - Several previous reports concluded that the C4b fragment of human C4A (C4Ab) binds with higher affinity to CR1 than does C4Bb. Because the isotypic residues, (1101)PCPVLD and (1101)LSPVIH in C4A and C4B, respectively, are located within the C4d region, one may have expected a direct binding contribution of C4d to the interaction with CR1. However, using surface plasmon resonance as our analytical tool, with soluble rCR1 immobilized on the biosensor chip, we failed to detect significant binding of C4d of either isotype. By contrast, binding of C4c was readily detectable. C4A and C4B, purified from plasma lacking one of the isotypes, were Cs converted to C4Ab and C4Bb. Spontaneously formed disulfide linked dimers were separated from monomers and higher oligomers by sequential chromatographic steps. The binding sensorgrams of C4Ab and C4Bb monomers as analytes reached steady state plateaus, and these equilibrium data yielded essentially superimposable saturation curves that were well fit by a one-site binding model. Although a two-site model was required to fit the equilibrium binding data for the dimeric forms of C4b, once again there was little difference in the K(D) values obtained for each isotype. Independent verification of our surface plasmon resonance studies came from ELISA-based inhibition experiments in which monomers of C4Ab and C4Bb were equipotent in inhibiting the binding of soluble CR1 to plate-bound C4b. Although divergent from previous reports, our results are consistent with recent C4Ad structural data that raised serious doubts about there being a conformational basis for the previously reported isotypic differences in the C4b-CR1 interaction. PMID- 14734750 TI - Hypoxia selectively inhibits monocyte chemoattractant protein-1 production by macrophages. AB - Hypoxia, a local decrease in oxygen tension occurring in inflammatory and tumor lesions, modulates gene expression in macrophages. Because macrophages are important chemokine producers, we investigated the regulatory effects of hypoxia on macrophage-derived chemokines. We demonstrated that hypoxia inhibits the production of the macrophage and T lymphocyte chemotactic and activating factor, monocyte chemoattractant protein-1 (MCP-1). Exposure of mouse macrophages to low oxygen tension resulted in the down-regulation of constitutive MCP-1 mRNA expression and protein secretion. Hypoxia inhibitory effects were selective for MCP-1 because the chemokines macrophage inflammatory protein-1beta (MIP-1beta), RANTES, IFN-gamma-inducible protein-10, and MIP-2 were not affected, and MIP 1alpha was induced. Hypoxia also inhibited, in a time-dependent fashion, MCP-1 up regulation by IFN-gamma and LPS. Moreover, the inhibitory action of hypoxia was exerted on human monocytic cells. MCP-1 down-regulation was associated with inhibition of gene transcription and mRNA destabilization, suggesting a dual molecular mechanism of control. Finally, we found that the triptophan catabolite picolinic acid and the iron chelator desferrioxamine, which mimic hypoxia in the induction of gene expression, differentially regulated the expression of MCP-1. This study characterizes a novel property of hypoxia as a selective inhibitor of MCP-1 production induced by different stimuli in macrophages and demonstrates that down-regulation of gene expression by hypoxia can be controlled at both transcriptional and posttranscriptional levels. Inhibition of MCP-1 may represent a negative regulatory mechanism to control macrophage-mediated leukocyte recruitment in pathological tissues. PMID- 14734751 TI - Critical role of OX40 in CD28 and CD154-independent rejection. AB - Blocking both CD28 and CD154 costimulatory pathways can induce transplant tolerance in some, but not all, transplant models. Under stringent conditions, however, this protocol often completely fails to block allograft rejection. The precise nature of such CD28/CD154 blockade-resistant rejection is largely unknown. In the present study we developed a new model in which both CD28 and CD154, two conventional T cell costimulatory molecules, are genetically knocked out (i.e., CD28/CD154 double-knockout (DKO) mice) and used this model to examine the role of novel costimulatory molecule-inducible costimulator (ICOS), OX40, 4 1BB, and CD27 in mediating CD28/CD154-independent rejection. We found that CD28/CD154 DKO mice vigorously rejected fully MHC-mismatched DBA/2 skin allografts (mean survival time, 12 days; n = 6) compared with the wild-type controls (mean survival time, 8 days; n = 7). OX40 costimulation is critically important in skin allograft rejection in this model, as blocking the OX40/OX40 ligand pathway, but not the ICOS/ICOS ligand, 4-1BB/4-1BBL, or CD27/CD70 pathway, markedly prolonged skin allograft survival in CD28/CD154 DKO mice. The critical role of OX40 costimulation in CD28/CD154-independent rejection is further confirmed in wild-type C57BL/6 mice, as blocking the OX40/OX40 ligand pathway in combination with CD28/CD154 blockade induced long term skin allograft survival (>100 days; n = 5). Our study revealed a key cellular mechanism of rejection and identified OX40 as a critical alternative costimulatory molecule in CD28/CD154 independent rejection. PMID- 14734752 TI - IFN-alpha sensitizes human umbilical vein endothelial cells to apoptosis induced by double-stranded RNA. AB - The ability of endothelial cells to mount an efficient antiviral response is important in restricting viral dissemination and eliminating viral infection from the endothelium and surrounding tissues. We demonstrate that dsRNA, a molecular signature of viral infection, induced apoptosis in HUVEC, and priming with IFN alpha shortened the time between when dsRNA was encountered and when apoptosis was initiated. IFN-alpha priming induced higher levels of mRNA for dsRNA activated protein kinase, 2'5'-oligoadenylate synthetase, and Toll-like receptor 3, transcripts that encode dsRNA-responsive proteins. dsRNA induced activation of dsRNA-activated protein kinase and nuclear translocation of transcription factors RelA and IFN regulatory factor-3 in IFN-alpha-primed HUVECs before the activation of intrinsic and extrinsic apoptotic pathways. These changes did not occur in the absence of dsRNA, and apoptosis resulting from incubation with dsRNA occurred much later when cells were not primed with IFN-alpha. The entire population of IFN-alpha-primed HUVECs underwent nuclear translocation of RelA and IFN regulatory factor-3 in response to dsRNA, whereas less than one-half of the population responded with apoptosis. When IFN-alpha-primed HUVECs were coincubated with dsRNA and proteasome inhibitors, all HUVECs were rendered susceptible to dsRNA-induced apoptosis. These studies provide evidence that many endothelial cells that are alerted to the risk of infection by IFN-alpha would undergo apoptosis sooner in response to dsRNA than non-IFN-alpha-primed cells, and this would enhance the likelihood of eliminating infected cells prior to the production of progeny virions. PMID- 14734753 TI - Impaired production of proinflammatory cytokines and host resistance to acute infection with Trypanosoma cruzi in mice lacking functional myeloid differentiation factor 88. AB - Studies performed in vitro suggest that activation of Toll-like receptors (TLRs) by parasite-derived molecules may initiate inflammatory responses and host innate defense mechanisms against Trypanosoma cruzi. Here, we evaluated the impact of TLR2 and myeloid differentiation factor 88 (MyD88) deficiencies in host resistance to infection with T. cruzi. Our results show that macrophages derived from TLR2 (-/-) and MyD88(-/-) mice are less responsive to GPI-mucin derived from T. cruzi trypomastigotes and parasites. In contrast, the same cells from TLR2(-/ ) still produce TNF-alpha, IL-12, and reactive nitrogen intermediates (RNI) upon exposure to live T. cruzi trypomastigotes. Consistently, we show that TLR2(-/-) mice mount a robust proinflammatory cytokine response as well as RNI production during the acute phase of infection with T. cruzi parasites. Further, deletion of the functional TLR2 gene had no major impact on parasitemia nor on mortality. In contrast, the MyD88(-/-) mice had a diminished cytokine response and RNI production upon acute infection with T. cruzi. More importantly, we show that MyD88(-/-) mice are more susceptible to infection with T. cruzi as indicated by the higher parasitemia and accelerated mortality, as compared with the wild-type mice. Together, our results indicate that T. cruzi parasites elicit an alternative inflammatory pathway independent of TLR2. This pathway is partially dependent on MyD88 and necessary for mounting optimal inflammatory and RNI responses that control T. cruzi replication during the early stages of infection. PMID- 14734754 TI - HLA class II polymorphisms determine responses to bacterial superantigens. AB - The excessive immunological response triggered by microbial superantigens has been implicated in the etiology of a wide range of human diseases but has been most clearly defined for the staphylococcal and streptococcal toxic shock syndromes. Because MHC class II presentation of superantigens to T cells is not MHC-restricted, the possibility that HLA polymorphisms could influence superantigenicity, and thus clinical susceptibility to the toxicity of individual superantigens, has received little attention. In this study, we demonstrate that binding of streptococcal and staphylococcal superantigens to HLA class II is influenced by allelic differences in class II. For the superantigen streptococcal pyrogenic exotoxin A, class II binding is dependent on DQ alpha-chain polymorphisms such that HLA-DQA1*01 alpha-chains show greater binding than DQA1*03/05 alpha-chains. The functional implications of differential binding on T cell activation were investigated in various experimental systems using human T cells and murine Vbeta8.2 transgenic cells as responders. These studies showed quantitative and qualitative differences resulting from differential HLA-DQ binding. We observed changes in T cell proliferation and cytokine production, and in the Vbeta specific changes in T cell repertoire that have hitherto been regarded as a defining feature of an individual superantigen. Our observations reveal a mechanism for the different outcomes seen following infection by toxigenic bacteria. PMID- 14734755 TI - Dendritic cells pulsed with live and dead Legionella pneumophila elicit distinct immune responses. AB - Legionella pneumophila is the causative pathogen of Legionnaires' disease, which is characterized by severe pneumonia. In regard to the pathophysiology of Legionella infection, the role of inflammatory phagocytes such as macrophages has been well documented, but the involvement of dendritic cells (DCs) has not been clarified. In this study, we have investigated the immune responses that DCs generate in vitro and in vivo after contact with L. pneumophila. Heat- and formalin-killed L. pneumophila, but not live L. pneumophila, induced immature DCs to undergo similar phenotypic maturation, but the secreted proinflammatory cytokines showed different patterns. The mechanisms of the DC maturation by heat- or formalin-killed L. pneumophila depended, at least in part, on Toll-like receptor 4 signaling or on Legionella LPS, respectively. After transfer to naive mice, DCs pulsed with dead Legionella produced serum Ig isotype responses specific for Legionella, leading to protective immunity against an otherwise lethal respiratory challenge with L. pneumophila. The in vivo immune responses required the Ag presentation of DCs, especially that on MHC class II molecules, and the immunity yielded cross-protection between clinical and environmental strains of L. pneumophila. Although the DC maturation was impaired by live Legionella, macrophages were activated by live as well as dead L. pneumophila, as evidenced by the up-regulation of MHC class II. Finally, DCs, but not macrophages, exhibited a proliferative response to live L. pneumophila that was consistent with their cell cycle progression. These findings provide a better understanding of the role of DCs in adaptive immunity to Legionella infection. PMID- 14734756 TI - Human T cell lymphotropic virus type I (HTLV-I)-specific CD4+ T cells: immunodominance hierarchy and preferential infection with HTLV-I. AB - CD4(+) T cells predominate in early lesions in the CNS in the inflammatory disease human lymphotropic T cell virus type I (HTLV-I)-associated myelopathy/tropical spastic paraparesis (HAM/TSP), but the pathogenesis of the disease remains unclear and the HTLV-I-specific CD4(+) T cell response has been little studied. We quantified the IFN-gamma-producing HTLV-I-specific CD4(+) T cells, in patients with HAM/TSP and in asymptomatic carriers with high proviral load, to test two hypotheses: that HAM/TSP patients and asymptomatic HTLV-I carriers with a similar proviral load differ in the immunodominance hierarchy or the total frequency of specific CD4(+) T cells, and that HTLV-I-specific CD4(+) T cells are preferentially infected with HTLV-I. The strongest CD4(+) T cell response in both HAM/TSP patients and asymptomatic carriers was specific to Env. This contrasts with the immunodominance of Tax in the HTLV-I-specific CD8(+) T cell response. The median frequency of HTLV-I-specific IFN-gamma(+) CD4(+) T cells was 25-fold greater in patients with HAM/TSP (p = 0.0023, Mann-Whitney) than in asymptomatic HTLV-I carriers with a similar proviral load. Furthermore, the frequency of CD4(+) T cells infected with HTLV-I (expressing Tax protein) was significantly greater (p = 0.0152, Mann-Whitney) among HTLV-I-specific cells than CMV-specific cells. These data were confirmed by quantitative PCR for HTLV-I DNA. We conclude that the high frequency of specific CD4(+) T cells was associated with the disease HAM/TSP, and did not simply reflect the higher proviral load that is usually found in HAM/TSP patients. Finally, we conclude that HTLV-I specific CD4(+) T cells are preferentially infected with HTLV-I. PMID- 14734757 TI - Pervasive influence of hepatitis C virus on the phenotype of antiviral CD8+ T cells. AB - Recent studies using MHC class I tetramers have shown that CD8(+) T cell responses against different persistent viruses vary considerably in magnitude and phenotype. At one extreme, hepatitis C virus (HCV)-specific CD8(+) T cell responses in blood are generally weak and have a phenotype that is perforin low and CCR7 high (early memory). At the other, specific responses to CMV are strong, perforin high, and CCR7 low (mature or effector memory). To examine the potential mechanisms behind this diversity, we compared CMV-specific responses in HCV infected and healthy individuals. We find a striking difference in the phenotype of CMV-specific CD8(+) T cells between these groups. In the HCV-infected cohort, CMV-specific CD8(+) T cells lost markers associated with maturity; they had increased expression of CCR7 and reduced expression of Fas and perforin. They nevertheless responded to Ag in vitro in a manner similar to controls, with strong proliferation and appropriate acquisition of effector memory markers. The reduction in mature CD8 T cells in HCV-infected individuals may arise through either impairment or regulation of T cell stimulation, or through the early loss of mature T cells. Whatever the mechanism, HCV has a pervasive influence on the circulating CD8(+) T cell population, a novel feature that may be a hallmark of this infection. PMID- 14734758 TI - Distinct modulatory effects of LPS and CpG on IL-18-dependent IFN-gamma synthesis. AB - Innate cellular production of IFN-gamma is suppressed after repeated exposure to LPS, whereas CpG-containing DNA potentiates IFN-gamma production. We compared the modulatory effects of LPS and CpG on specific cellular and cytokine responses necessary for NK-cell dependent IFN-gamma synthesis. C3H/HeN mice pretreated with LPS for 2 days generated 5-fold less circulating IL-12 p70 and IFN-gamma in response to subsequent LPS challenge than did challenged control mice. In contrast, CpG-pretreated mice produced 10-fold more circulating IFN-gamma without similar changes in IL-12 p70 levels, but with 10-fold increases in serum IL-18 relative to LPS-challenged control or endotoxin-tolerant mice. The role of IL-18 in CpG-induced immune potentiation was studied in splenocyte cultures from control, LPS-conditioned, or CpG-conditioned mice. These cultures produced similar amounts of IFN-gamma in response to rIL-12 and rIL-18. However, only CpG conditioned cells produced IFN-gamma when cultured with LPS or CpG, and production was ablated in the presence of anti-IL-18R Ab. Anti-IL-18R Ab also reduced in vivo IFN-gamma production by >2-fold in CpG-pretreated mice. Finally, combined pretreatment of mice with LPS and CpG suppressed the production of circulating IFN-gamma, IL-12 p70, and IL-18 after subsequent LPS challenge. We conclude that CpG potentiates innate IFN-gamma production from NK cells by increasing IL-18 availability, but that the suppressive effects of LPS on innate cellular immunity dominate during combined LPS and CpG pretreatment. Multiple Toll-like receptor engagement in vivo during infection can result in functional polarization of innate immunity dominated by a specific Toll-like receptor response. PMID- 14734759 TI - Selective killing of vaccinia virus by LL-37: implications for eczema vaccinatum. AB - Possible bioterrorism with smallpox has led to the resumption of smallpox (vaccinia virus) immunization. One complication, eczema vaccinatum, occurs primarily in patients with atopic dermatitis (AD). Skin lesions of patients with AD, but not psoriasis, is deficient in the cathelicidin antimicrobial peptide (LL 37) and human beta-defensin-2 (HBD-2). We hypothesized that this defect may explain the susceptibility of patients with AD to eczema vaccinatum. The Wyeth vaccine strain of vaccinia virus was incubated with varying concentrations of human (LL-37) and murine (CRAMP) cathelicidins, human alpha-defensin (HBD-1, HBD 2), and a control peptide. Outcomes included quantification of viral PFU, vaccinia viral gene expression by quantitative real-time RT-PCR, and changes in virion structure by transmission electron microscopy. CRAMP knockout mice and control animals were inoculated by skin pricks with 2 x 10(5) PFU of vaccinia and examined daily for pox development. Physiologic amounts of human and murine cathelicidins (10-50 micro M), but not human defensins, which had antibacterial activity, resulted in the in vitro reduction of vaccinia viral plaque formation (p < 0.0001), vaccinia mRNA expression (p < 0.001), and alteration of vaccinia virion structure. In vivo vaccinia pox formation occurred in four of six CRAMP knockout animals and in only one of 15 control mice (p < 0.01). These data support a role for cathelicidins in the inhibition of orthopox virus (vaccinia) replication both in vitro and in vivo. Susceptibility of patients with AD to eczema vaccinatum may be due to a deficiency of cathelicidin. PMID- 14734760 TI - Chlamydia pneumoniae multiply in neutrophil granulocytes and delay their spontaneous apoptosis. AB - The obligate intracellular bacterial pathogen Chlamydia pneumoniae (Cp) is responsible for a range of human diseases, including acute respiratory infection. Although experimental intratracheal infection with Cp results in a massive recruitment of neutrophil granulocytes (polymorphonuclear neutrophils (PMN)), the role of these cells in the defense against Cp is unclear. In this study the interactions of PMN with Cp were investigated. In vitro coincubation experiments showed that human granulocytes were able to internalize Chlamydia in an opsonin independent manner. Importantly, phagocytosed Cp were not killed; the ingested bacteria survived and multiplied within PMN. Although uninfected granulocytes became apoptotic within 10 h, infected PMN survived up to 90 h. Coincubation with Cp significantly decreased the ratio of apoptotic PMN, as detected by morphological analysis, annexin V, and TUNEL staining. The observed antiapoptotic effect was associated with a markedly lower level of procaspase-3 processing and, consequently, reduced caspase-3 activity in infected PMN. LPS was found as a major, but not exclusive, component responsible for the observed antiapoptotic effect. Chlamydia LPS affected PMN apoptosis both by acting directly on the cells and by inducing the autocrine production of the antiapoptotic cytokine IL-8. These data show that, in contrast to other microbial pathogens that drive phagocytes into apoptosis to escape killing, Cp can extend the life span of neutrophil granulocytes, making them suitable host cells for survival and multiplication within the first hours/days after infection. PMID- 14734761 TI - Nonmethylated CG motifs packaged into virus-like particles induce protective cytotoxic T cell responses in the absence of systemic side effects. AB - DNA rich in nonmethylated CG motifs (CpGs) greatly facilitates induction of immune responses against coadministered Ags. CpGs are therefore among the most promising adjuvants known to date. Nevertheless, CpGs are characterized by two drawbacks. They have unfavorable pharmacokinetics and may exhibit systemic side effects, including splenomegaly. We show in this study that packaging CpGs into virus-like particles (VLPs) derived from the hepatitis B core Ag or the bacteriophage Qbeta is a simple and attractive method to reduce these two problems. CpGs packaged into VLPs are resistant to DNase I digestion, enhancing their stability. In addition, and in contrast to free CpGs, packaging CpGs prevents splenomegaly in mice, without affecting their immunostimulatory capacity. In fact, vaccination with CpG-loaded VLPs was able to induce high frequencies of peptide-specific CD8(+) T cells (4-14%), protected from infection with recombinant vaccinia viruses, and eradicated established solid fibrosarcoma tumors. Thus, packaging CpGs into VLPs improves both their immunogenicity and pharmacodynamics. PMID- 14734762 TI - Overproduction of TNF-alpha by CD8+ type 1 cells and down-regulation of IFN-gamma production by CD4+ Th1 cells contribute to toxic shock-like syndrome in an animal model of fatal monocytotropic ehrlichiosis. AB - Human monocytotropic ehrlichiosis (HME) is an emerging, life-threatening, infectious disease caused by Ehrlichia chaffeensis, an obligate intracellular bacterium that lacks cell wall LPS. We have previously developed an animal model of severe HME using a strain of Ehrlichia isolated from Ixodes ovatus ticks (IOE). To understand the basis of susceptibility to severe monocytotropic ehrlichiosis, we compared low and high doses of the highly virulent IOE strain and the less virulent Ehrlichia muris strain that are closely related to E. chaffeensis in C57BL/6 mice. Lethal infections caused by high or low doses of IOE were accompanied by extensive liver damage, extremely elevated levels of TNF alpha in the serum, high frequency of Ehrlichia-specific, TNF-alpha-producing CD8(+) T cells in the spleen, decreased Ehrlicha-specific CD4(+) T cell proliferation, low IL-12 levels in the spleen, and a 40-fold decrease in the number of IFN-gamma-producing CD4(+) Th1 cells. All groups contained negligible numbers of IL-4-producing cells in the spleen. Transfer of Ehrlichia-specific polyclonal Abs and IFN-gamma-producing Ehrlichia-specific CD4(+) and CD8(+) type 1 cells protected naive mice against lethal IOE challenge. Interestingly, infection with high dose E. muris provided protection against rechallenge with a lethal dose of IOE. Cross-protection was associated with substantial expansion of IFN-gamma-producing CD4(+) and CD8(+) cells, but not TNF-alpha-producing CD8(+) T cells, a high titer of IgG2a, and a low serum level of TNF-alpha. In conclusion, uncontrolled TNF-alpha production by CD8(+) T cells together with a weak CD4(+) Th1 cell response are associated with immunopathology and failure to clear IOE in the fatal model of HME. PMID- 14734764 TI - Leptin indirectly activates human neutrophils via induction of TNF-alpha. AB - Leptin, the satiety hormone, appears to act as a link between nutritional status and immune function. It has been shown to elicit a number of immunoregulatory effects, including the promotion of T cell proliferative responses, and the induction of proinflammatory cytokines. Leptin deficiency is associated with an increased susceptibility to infection. As polymorphonuclear neutrophils (PMN) play a major role in innate immunity and host defense against infection, this study evaluated the influence of leptin on PMN activation. The presence of leptin receptor in human PMN was determined both at mRNA and protein levels, and the effect of leptin on PMN activation, as assessed by CD11b expression, was evaluated using flow cytometry. In contrast to monocytes, which express both the short and long forms of the leptin receptor (Ob-Ra and Ob-Rb, respectively), PMN expressed only Ob-Ra. Leptin up-regulated the expression of CD11b, an early marker of PMN activation, on PMN in whole blood, yet it had no effect on purified PMN, even those treated by submaximal doses of TNF-alpha or PMA. The kinetics of leptin-induced activation in whole blood were consistent with an indirect effect mediated by monocytes, and 71% of the leptin-stimulatory effect on PMN was blocked by a TNF-alpha inhibitor. Leptin-mediated induction of CD11b expression was observed when purified PMN were coincubated with purified monocytes. In conclusion, although leptin activates PMN, it does so indirectly via TNF-alpha release from monocytes. These findings provide an additional link among the obesity-derived hormone leptin, innate immune function, and infectious disease. PMID- 14734763 TI - p47phox deficiency impairs NF-kappa B activation and host defense in Pseudomonas pneumonia. AB - We examined the role of redox signaling generated by NADPH oxidase in activation of NF-kappaB and host defense against Pseudomonas aeruginosa pneumonia. Using mice with an NF-kappaB-driven luciferase reporter construct (HIV-LTR/luciferase (HLL)), we found that intratracheal administration of P. aeruginosa resulted in a dose-dependent neutrophilic influx and activation of NF-kappaB. To determine the effects of reactive oxygen species generated by the NADPH oxidase system on activation of NF-kappaB, we crossbred mice deficient in p47(phox) with NF-kappaB reporter mice (p47(phox-/-)HLL). These p47(phox-/-)HLL mice were unable to activate NF-kappaB to the same degree as HLL mice with intact NADPH oxidase following P. aeruginosa infection. In addition, lung TNF-alpha levels were significantly lower in p47(phox-/-)HLL mice compared with HLL mice. Bacterial clearance was impaired in p47(phox-/-)HLL mice. In vitro studies using bone marrow-derived macrophages showed that Toll-like receptor 4 was necessary for NF kappaB activation following treatment with P. aeruginosa. Additional studies with macrophages from p47(phox-/-) mice confirmed that redox signaling was necessary for maximal Toll-like receptor 4-dependent NF-kappaB activation in this model. These data indicate that the NADPH oxidase-dependent respiratory burst stimulated by Pseudomonas infection contributes to host defense by modulating redox dependent signaling through the NF-kappaB pathway. PMID- 14734765 TI - Transcript signatures in experimental asthma: identification of STAT6-dependent and -independent pathways. AB - The analysis of polygenic diseases such as asthma poses a challenging problem. In an effort to provide unbiased insight into disease pathogenesis, we took an empirical approach involving transcript expression profiling of lung tissue from mice with experimental asthma. Asthmatic responses were found to involve sequential induction of 4.7% of the tested genome; notably, there was ectopic expression of a series of genes not previously implicated in allergic or pulmonary responses. Genes were widely distributed throughout all chromosomes, but preferentially included genes involved in immunity, development, and homeostasis. When asthma was induced by two independent experimental regimens, unique gene transcript profiles were found depending upon the mode of disease induction. However, the majority of genes were common to both models representing an asthma signature genome. Analysis of STAT6-deficient mice revealed that an unexpectedly large segment of the asthma genes were STAT6 independent; this correlated with sustained inflammatory events in these mice. Notably, induction of asthma in STAT6-deficient mice resulted in gene induction not seen in wild type mice. These results raise concern that therapeutic blockade of STAT6 in the asthmatic setting may reprogram the genetic signature, resulting in alternative lung pathology, which we indeed observed in STAT6-deficient mice. These results provide unprecedented insight into the complex steps involved in the pathogenesis of allergic airway responses; as such, these results have significant therapeutic and clinical implications. PMID- 14734766 TI - Macrophage-stimulating protein, the ligand for the stem cell-derived tyrosine kinase/RON receptor tyrosine kinase, inhibits IL-12 production by primary peritoneal macrophages stimulated with IFN-gamma and lipopolysaccharide. AB - IL-12, produced by APCs during the initial stages of an immune response, plays a pivotal role in the induction of IFN-gamma by NK and gammadeltaT cells and in driving the differentiation of Th1 cells, thus providing a critical link between innate and acquired immunity. Due to the unique position occupied by IL-12 in the regulation of immunity, many mechanisms have evolved to modulate IL-12 production. We have shown previously that macrophage-stimulating protein (MSP), the ligand for the stem cell-derived tyrosine kinase/recepteur d'origine nantais (RON) receptor, inhibits NO production by macrophages in response to IFN-gamma and enhances the expression of arginase. Mice lacking RON exhibit increased inflammation in a delayed-type hypersensitivity reaction and increased susceptibility to endotoxic shock. In this study we demonstrate that pretreatment of macrophages with MSP before IFN-gamma and LPS results in the complete inhibition of IL-12 production due to suppression of p40 expression. This response is mediated by the RON receptor, and splenocytes from RON(-/-) animals produce increased levels of IFN-gamma. MSP pretreatment of macrophages resulted in decreased tyrosine phosphorylation of Stat-1 and decreased expression of IFN consensus sequence binding protein in response to inflammatory cytokines. In addition to IL-12, the expression of IL-15 and IL-18, cytokines that are also dependent on IFN consensus sequence binding protein activation, is inhibited by pretreatment with MSP before IFN-gamma and LPS. We also show that the ability of MSP to inhibit IL-12 production is independent of IL-10. Taken together, these results suggest that MSP may actively suppress cell-mediated immune responses through its ability to down-regulate IL-12 production and thus inhibit classical activation of macrophages. PMID- 14734767 TI - Cysteinyl leukotrienes regulate dendritic cell functions in a murine model of asthma. AB - Dendritic cells (DCs) act as APCs in the airway and play a critical role in allergy. Cysteinyl leukotrienes (cysLTs) synthesized from arachidonic acid are primary mediators of immediate asthmatic reaction. The aim of this study was to investigate the effects of cysLTs on Dermatophagoides farinae (Der f)-pulsed mouse myeloid DCs in inducing allergic airway inflammation in vitro and in vivo. Control DC (medium-pulsed), Der f-pulsed DC, cysLT-pulsed DC, Der f- and cysLT pulsed DC, and Der f-pulsed and cysLT receptor antagonist (LTRA)-treated DC were prepared from murine bone marrow, and the production of cytokines ws compared. Subsequently, these DCs were intranasally instilled into another group of naive mice, followed by intranasal Der f challenge to induce allergic airway inflammation in vivo. Der f-pulsed DC produced significantly higher amounts of IL 10 and IL-12 compared with control DC. Der f- and cysLT-pulsed DC further increased IL-10 production compared with Der f-pulsed DC. In contrast, treatment of Der f-pulsed DC with LTRA increased IL-12 and decreased IL-10. Intranasal instillation of Der f-pulsed DC resulted in airway eosinophilia associated with a significant rise in IL-5 levels in the airway compared with control DC. Pulmonary eosinophilia and excess IL-5 were further enhanced in Der f- and cysLT-pulsed DC harboring mice. In contrast, Der f-pulsed and LTRA-treated DC significantly inhibited airway eosinophilia, reduced IL-5, and increased IFN-gamma in the airway. Our results suggest that cysLTs play an important role in the development of allergic airway inflammation by regulating the immunomodulatory functions of DCs. PMID- 14734768 TI - Kinin B1 receptor up-regulation after lipopolysaccharide administration: role of proinflammatory cytokines and neutrophil influx. AB - Several studies have now clearly established the ability of LPS to induce bradykinin B(1) receptor up-regulation in vivo and the functional relevance of this up-regulation for the pathophysiological effects of LPS. Using an in vivo system in which LPS is injected locally into the rat paw, we have examined the potential contribution of proinflammatory cytokines, NF-kappaB activation, and neutrophil influx for the functional and molecular up-regulation of the bradykinin B(1) receptor. Treatment with LPS resulted in a rapid and sustained functional up-regulation of B(1) receptors in the rat paw that correlated with the increase in B(1) receptor mRNA levels. B(1) receptor up-regulation is preceded by the rapid activation of the transcription factor NF-kappaB and the production of proinflammatory cytokines, including TNF-alpha and IL-1beta. More importantly, blockade of NF-kappaB translocation, TNF-alpha, or IL-1beta prevented the functional and molecular up-regulation of B(1) receptors. Injection of LPS also induced the influx of neutrophils that followed the peak of cytokine production and associated with the persistent activation of NF-kappaB and functional B(1) receptor up-regulation. Blockade of neutrophil influx with platelet-activating factor receptor antagonists or cell adhesion molecule blockers prevented B(1) receptor up-regulation. Thus, by acting in cooperation and in a coordinated, timely manner, TNF-alpha, IL-1beta, neutrophils, and the transcription factor NF-kappaB are major and essential players in the ability of LPS to induce B(1) receptor expression in vivo. PMID- 14734769 TI - The essential involvement of cross-talk between IFN-gamma and TGF-beta in the skin wound-healing process. AB - Several lines of in vitro evidence suggest the potential role of IFN-gamma in angiogenesis and collagen deposition, two crucial steps in the wound healing process. In this report, we examined the role of IFN-gamma in the skin wound healing process utilizing WT and IFN-gamma KO mice. In WT mice, excisional wounding induced IFN-gamma mRNA and protein expression by infiltrating macrophages and T cells, with a concomitant enhancement of IL-12 and IL-18 gene expression. Compared with WT mice, IFN-gamma KO mice exhibited an accelerated wound healing as evidenced by rapid wound closure and granulation tissue formation. Moreover, IFN-gamma KO mice exhibited enhanced angiogenesis with augmented vascular endothelial growth factor mRNA expression in wound sites, compared with WT mice, despite a reduction in the infiltrating neutrophils, macrophages, and T cells. IFN-gamma KO mice also exhibited accelerated collagen deposition with enhanced production of TGF-beta1 protein in wound sites, compared with WT mice. Furthermore, the absence of IFN-gamma augmented the TGF-beta1 mediated signaling pathway, as evidenced by increases in the levels of total and phosphorylated Smad2 and a reciprocal decrease in the levels of Smad7. These results demonstrate that there is crosstalk between the IFN-gamma/Stat1 and TGF beta1/Smad signaling pathways in the wound healing process. PMID- 14734771 TI - Expression of CD154 (CD40 ligand) by human lung fibroblasts: differential regulation by IFN-gamma and IL-13, and implications for fibrosis. AB - The CD40-CD40 ligand (CD40L) system (CD154) is a central means of immune cell communication crucial for Ig class switching and enhanced Ag presentation. CD40 is also a key signaling conduit to activate nonhematopoietic cells, such as fibroblasts and endothelial cells, to produce proinflammatory mediators. Disruption of the CD40-CD40L pathway reduces lung inflammation and fibrosis, autoimmune disease and atherosclerosis. Non-bone marrow-derived structural cells are not known to express CD40L. In this study, we reveal the intriguing finding that primary strains of human lung fibroblasts derived from normal and scarred lung express both CD40L mRNA and protein. Interestingly, CD40L expression is down regulated by IFN-gamma, a type 1 cytokine with antiscarring properties, and is up regulated by the profibrogenic type 2 cytokine IL-13. Flow cytometry and laser confocal microscopy revealed that the majority of CD40L was located intracellularly. Importantly, fibroblast strains from human idiopathic pulmonary fibrosis tissue expressed increased levels of CD40L compared with fibroblasts from nonscarred lung. Fibroblasts in the scarred areas of human lung tissue expressed high levels of CD40L. Finally, the blood and lung lavage levels of CD40L are significantly elevated in fibrosis patients compared with normals. These new findings demonstrate that fibroblasts are a new source of CD40L and that those involved in scarring may have undergone a selected expansion for high CD40L expression. Moreover, the antifibrotic activity of IFN-gamma may involve the down-regulation of fibroblast CD40L levels. We speculate that fibroblast derived CD40L plays a role in promoting fibroblast activation and possibly in interaction with CD40 bearing cells. PMID- 14734770 TI - A molecular defect in intracellular lipid signaling in human neutrophils in localized aggressive periodontal tissue damage. AB - Host defense mechanisms are impaired in patients with congenital neutrophil (polymorphonuclear neutrophils (PMN)) defects. Impaired PMN chemotaxis is observed in localized aggressive periodontitis (LAP), a familial disorder characterized by destruction of the supporting structures of dentition. In the present studies, we sought evidence for molecular events underlying this aberrant human PMN phenotype. To this end, PMN transendothelial migration and superoxide anion generation were assessed with LAP patients and asymptomatic family members, as well as patients with other chronic mucosal inflammation. PMN from LAP patients showed decreased transmigration across vascular endothelial monolayers (18 +/- 12% of control, n = 4) and increased superoxide anion generation (358 +/- 37%, p = 0.003). Gene expression was analyzed using oligonucleotide microarrays and fluorescence-based kinetic PCR. cDNA microarray and kinetic-PCR analysis revealed diminished RNA expression of leukocyte-type diacylglycerol (DAG) kinase alpha in PMN from LAP patients (4.6 +/- 1.7 relative units, n = 6, p = 0.007) compared with asymptomatic individuals (51 +/- 27 relative units, n = 7). DAG kinase activity was monitored by DAG phosphorylation and individual DAG molecular species were quantified using liquid chromatography and tandem mass spectrometry based lipidomics. DAG kinase activity was also significantly decreased (73 +/- 2%, p = 0.007) and correlated with increased accumulation of 1,2-diacyl-sn-3 glycerol substrates (p = 0.01). These results implicate defects in both PMN transendothelial migration and PMN DAG kinase alpha signaling as disordered functions in LAP. Moreover, they identify a potential molecular lesion in PMN signal transduction that may account for their aberrant responses and tissue destruction in this disease. PMID- 14734772 TI - The C10/CCL6 chemokine and CCR1 play critical roles in the pathogenesis of IL-13 induced inflammation and remodeling. AB - IL-13 is a potent stimulator of inflammation and tissue remodeling that plays a key role in the pathogenesis of a wide variety of human disorders. To further understand these responses, studies were undertaken to define the role(s) of the chemokine C10/CCL6 in the pathogenesis of IL-13-induced alterations in the murine lung. IL-13 was a very potent stimulator of C10/CCL6 mRNA and protein, and IL-13 induced inflammation, alveolar remodeling, and compliance alterations were markedly ameliorated after C10/CCL6 neutralization. Treatment with anti-C10/CCL6 decreased the levels of mRNA encoding matrix metalloproteinase-2 (MMP-2), MMP-9, and tissue inhibitor of metalloproteinase-4 (TIMP-4) in lungs from wild-type mice. C10/CCL6 neutralization also decreased the ability of IL-13 to stimulate the production of monocyte chemoattractant protein-1, macrophage inflammatory protein-1alpha, MMP-2, MMP-9, and cathepsins-K, -L, and -S and the ability of IL 13 to inhibit alpha1-antitrypsin. In accord with these findings, a targeted null mutation of CCR1, a putative C10/CCL6 receptor, also decreased IL-13-induced inflammation and alveolar remodeling and caused alterations in chemokines, proteases, and antiproteases comparable to those seen after C10/CCL6 neutralization. These C10/CCL6 and CCR1 manipulations did not alter the production of transgenic IL-13. These studies demonstrate that IL-13 is a potent stimulator of C10/CCL6 and highlight the importance of C10/CCL6 and signaling via CCR1 in the pathogenesis of the IL-13-induced pulmonary phenotype. They also describe a C10/CCL6 target gene cascade in which C10/CCL6 induction is required for optimal IL-13 stimulation of selected chemokines (monocyte chemoattractant protein-1 and MIP-1alpha) and proteases (MMP-2, MMP-9, and cathepsins-K, -L, and S) and the inhibition of alpha1-antitrypsin. PMID- 14734773 TI - Immune complexes bind preferentially to Fc gamma RIIA (CD32) on apoptotic neutrophils, leading to augmented phagocytosis by macrophages and release of proinflammatory cytokines. AB - Many human inflammatory diseases are associated with tissue deposition of immune complexes and influx of neutrophils. We show that immune complexes bind preferentially to apoptotic neutrophils via FcgammaRIIA (CD32) and that increased binding is associated with clustering of immune complexes on the plasma membrane of the apoptotic cell. Phagocytosis of immune complex-opsonized apoptotic neutrophils by human macrophages was substantially enhanced (4.4-fold increase compared with control apoptotic neutrophils) and stimulated macrophages to release the proinflammatory cytokines TNF-alpha and IL-6. Immune complexes may perturb the normal pathways for clearance of apoptotic neutrophils by augmenting their clearance at the price of proinflammatory cytokine release. This represents a novel mechanism by which immune complexes may modulate the resolution of inflammation. PMID- 14734774 TI - Induction of C chemokine XCL1 (lymphotactin/single C motif-1 alpha/activation induced, T cell-derived and chemokine-related cytokine) expression by HIV-1 Tat protein. AB - HIV-1 Tat has been proposed as a key agent in many AIDS-related disorders, including HIV-1-associated neurological diseases. We have recently shown that Tat expression induces a significant increase in T lymphocytes in the brains of Tat transgenic mice. The CNS infiltration of T lymphocytes has been noted in AIDS patients. In the present study using this unique genetic system we attempted to understand the underlying mechanisms of Tat expression-induced infiltration of T lymphocytes by examining chemokine expression. RNase protection assay revealed that in addition to CCL2 (monocyte chemoattractant protein-1), CCL3 (macrophage inflammatory protein-1alpha (MIP-1alpha)), CCL4 (MIP-1beta), CCL5 (RANTES), CXCL2 (MIP-2), and CXCL10 (inducing protein-10), XCL1 (lymphotactin/single C motif 1alpha/activation-induced, T cell-derived and chemokine-related cytokine) was identified to be up-regulated by Tat expression. XCL1 is a C chemokine and plays a specific and important role in tissue-specific recruitment of T lymphocytes. Thus, we further determined the relationship between Tat and XCL1 expression. Tat induced XCL1 expression was further confirmed by XCL1-specific RT-PCR and ELISA. Combined in situ hybridization and immunohistochemical staining identified astrocytes, monocytes, and macrophages/microglia as XCL1-producing cells in vivo. Using human astrocytes, U87.MG cells, as an in vitro model, activation of XCL1 expression was positively correlated with Tat expression. Moreover, the XCL1 promoter-driven reporter gene assay showed that Tat-induced XCL1 expression occurred at the transcriptional level. Taken together, these results demonstrate that Tat directly trans-activated XCL1 expression and suggest potential roles of Tat-induced XCL1 expression in the CNS infiltration of T lymphocytes during HIV-1 infection and subsequent HIV-1-induced neurological diseases. PMID- 14734775 TI - Chemotaxis and calcium responses of phagocytes to formyl peptide receptor ligands is differentially regulated by cyclic ADP ribose. AB - Cyclic ADP ribose (cADPR) is a calcium-mobilizing metabolite that regulates intracellular calcium release and extracellular calcium influx. Although the role of cADPR in modulating calcium mobilization has been extensively examined, its potential role in regulating immunologic responses is less well understood. We previously reported that cADPR, produced by the ADP-ribosyl cyclase, CD38, controls calcium influx and chemotaxis of murine neutrophils responding to fMLF, a peptide agonist for two chemoattractant receptor subtypes, formyl peptide receptor and formyl peptide receptor-like 1. In this study, we examine whether cADPR is required for chemotaxis of human monocytes and neutrophils to a diverse array of chemoattractants. We found that a cADPR antagonist and a CD38 substrate analogue inhibited the chemotaxis of human phagocytic cells to a number of formyl peptide receptor-like 1-specific ligands but had no effect on the chemotactic response of these cells to ligands selective for formyl peptide receptor. In addition, we show that the cADPR antagonist blocks the chemotaxis of human monocytes to CXCR4, CCR1, and CCR5 ligands. In all cases, we found that cADPR modulates intracellular free calcium levels in cells activated by chemokines that induce extracellular calcium influx in the apparent absence of significant intracellular calcium release. Thus, cADPR regulates calcium signaling of a discrete subset of chemoattractant receptors expressed by human leukocytes. Since many of the chemoattractant receptors regulated by cADPR bind to ligands that are associated with clinical pathology, cADPR and CD38 represent novel drug targets with potential application in chronic inflammatory and neurodegenerative disease. PMID- 14734777 TI - Lymphoid hyperplasia resulting in immune dysregulation is caused by porcine reproductive and respiratory syndrome virus infection in neonatal pigs. AB - Amid growing evidence that numerous viral infections can produce immunopathology, including nonspecific polyclonal lymphocyte activation, the need to test the direct impact of an infecting virus on the immune system of the host is crucial. This can best be tested in the isolator piglet model in which maternal and other extrinsic influences can be excluded. Therefore, neonatal isolator piglets were colonized with a benign Escherichia coli, or kept germfree, and then inoculated with wild-type porcine reproductive and respiratory syndrome virus (PRRSV) or sham medium. Two weeks after inoculation, serum IgM, IgG, and IgA levels were 30- to 50-, 20- to 80-, and 10- to 20-fold higher, respectively, in animals receiving virus vs sham controls, although <1% was virus specific. PRRSV-infected piglets also had bronchial tree-associated lymph nodes and submandibular lymph nodes that were 5-10 times larger than colonized, sham-inoculated animals. Size-exclusion fast performance liquid chromatography revealed that PRRSV-infected sera contained high-molecular-mass fractions that contained IgG, suggesting the presence of immune complexes. Lesions, inflammatory cell infiltration, glomerular deposits of IgG, IgM, and IgA, and Abs of all three isotypes to basement membrane and vascular endothelium were observed in the kidneys of PRRSV-infected piglets. Furthermore, autoantibodies specific for Golgi Ags and dsDNA could be detected 3 4 wk after viral inoculation. These data demonstrate that PRRSV induces B cell hyperplasia in isolator piglets that leads to immunologic injury and suggests that the isolator piglet model could serve as a useful model to determine the mechanisms of virus-induced immunopathology in this species. PMID- 14734776 TI - TNF-alpha-induced apoptosis of macrophages following inhibition of NF-kappa B: a central role for disruption of mitochondria. AB - Previously, we established that suppressing the constitutive activation of NF kappaB in in vitro matured human macrophages resulted in apoptosis initiated by a decrease of the Bcl-2 family member, A1, and the loss of mitochondrial transmembrane potential (Deltapsi(m)). This study was performed to characterize the mechanism of TNF-alpha-induced apoptosis in macrophages following the inhibition of NF-kappaB. The addition of TNF-alpha markedly enhanced the loss of Deltapsi(m) and the induction of apoptotic cell death. Although caspase 8 was activated and contributed to DNA fragmentation, it was not necessary for the TNF alpha-induced loss of Deltapsi(m). The inhibition of NF-kappaB alone resulted in the release of cytochrome c from the mitochondria, while both cytochrome c and second mitochondria-derived activator of caspase/direct inhibitor of apoptosis binding protein with low pI were released following the addition of TNF-alpha. Furthermore, c-Jun N-terminal kinase activation, which was sustained following treatment with TNF-alpha when NF-kappaB was inhibited, contributed to DNA fragmentation. These observations demonstrate that cytochrome c and second mitochondria-derived activator of caspase/direct inhibitor of apoptosis-binding protein with low pI may be differentially released from the mitochondria, and that the sustained activation of c-Jun N-terminal kinase modulated the DNA fragmentation independent of the loss of Deltapsi(m). PMID- 14734778 TI - Metal-protein complex-mediated transport and delivery of Ni2+ to TCR/MHC contact sites in nickel-specific human T cell activation. AB - Nickel allergy clearly involves the activation of HLA-restricted, skin-homing, Ni specific T cells by professional APCs. Nevertheless, knowledge concerning the molecular details of metal-protein interactions underlying the transport and delivery of metal ions to APC during the early sensitization phase and their interactions with HLA and TCRs is still fragmentary. This study investigates the role of human serum albumin (HSA), a known shuttling molecule for Ni(2+) and an often-disregarded, major component of skin, in these processes. We show that Ni saturated HSA complexes (HSA-Ni) induce and activate Ni-specific human T cells as potently as Ni salt solutions when present at equimolar concentrations classically used for in vitro T cell stimulation. However, neither HSA itself nor its Ni-binding N-terminal peptide are involved in determining the specificity of antigenic determinants. In fact, HSA could be replaced by xenogeneic albumins exhibiting sufficient affinity for Ni(2+) as determined by surface plasmon resonance (Biacore technology) or atomic absorption spectroscopy. Moreover, despite rapid internalization of HSA-Ni by APC, it was not processed into HLA associated epitopes recognizable by Ni-specific T cells. In contrast, the presence of HSA-Ni in the vicinity of transient contacts between TCR and APC exposed HLA molecules appeared to facilitate a specific transfer of Ni(2+) from HSA to high-affinity coordination sites created at the TCR/HLA-interface. PMID- 14734779 TI - Nucleotide sequencing of psoriatic arthritis tissue before and during methotrexate administration reveals a complex inflammatory T cell infiltrate with very few clones exhibiting features that suggest they drive the inflammatory process by recognizing autoantigens. AB - Psoriatic arthritis is an interesting MHC class I allele associated autoimmune disease where injury is likely mediated exclusively by T cells. We used TCR beta chain nucleotide sequencing to gain insight into the adaptive immune events responsible for this injury and determine whether the numerous oligoclonal expansions of this disease represent extreme determinant spreading among driving clones that recognize autoantigen or were non-Ag-driven, inflammation-related expansions. Because methotrexate suppresses but does not eliminate this inflammation, we hypothesized that clones persisting during methotrexate treatment would likely drive the inflammation. Seventy-six percent of the T cell clones in active tissue were polyclonal and unexpanded, accounting for 31% of transcripts. They were decreased greatly by methotrexate. Strikingly, most expanded clones in the inflamed joint did not persist during methotrexate treatment, were found only in inflammatory sites, exhibited no structural homology to one another, and were either CD4 or CD8 in lineage, suggesting they were non-autoantigen-driven, inflammation-related expansions. Only 12% of the expanded clones could be grouped into clonal sets distinguished by structurally homologous CDR3 beta-chain amino acid motifs suggesting Ag drive. These were exclusively CD8 in lineage, persisted during methotrexate administration, and were present in both joint fluid and blood implying they were candidate driver clones that recognized an autoantigen. However, a major set of putative driver clones exhibited a previously described EBV-specific beta-chain motif, emphasizing that the dominant feature of the disease was activation of multiple clones apparently lacking specificity for an inciting autoantigen. PMID- 14734780 TI - Ets-1 regulates TNF-alpha-induced matrix metalloproteinase-9 and tenascin expression in primary bronchial fibroblasts. AB - Increased subepithelial deposition of extracellular matrix proteins is a key feature in bronchial asthma. Matrix metalloproteinase-9 (MMP-9) is a proteolytic enzyme that degrades the extracellular matrix. Tenascin is an extracellular matrix glycoprotein that is abundant in thickened asthmatic subbasement membrane. The expression of MMP-9 and tenascin reflects disease activity in asthma and airway remodeling. The molecular mechanisms regulating the expression of these proteins remain unknown. Both MMP-9 and tenascin promoters contain an Ets binding site, suggesting control by Ets-1. Thus, we hypothesized that Ets-1 expression is increased in asthma and that it contributed to enhanced MMP-9 and tenascin expression. To test this hypothesis, we determined the expression of Ets-1 in bronchial biopsies obtained from asthmatic subjects and determined the expression of Ets-1, MMP-9, and tenascin by bronchial fibroblasts activated ex vivo. We observed that nuclear extracts from TNF-alpha-activated fibroblasts showed increased Ets-binding activity. In addition, TNF-alpha-activated fibroblasts had increased expression of Ets-1 mRNA and protein, which preceded an increase in MMP 9 and tenascin mRNA. Furthermore, treatment of fibroblasts with Ets-1 antisense oligonucleotides down-regulated TNF-alpha-induced Ets-1, MMP-9, and, to a lesser extent, tenascin protein expression or activity. Taken together, these data demonstrate that TNF-alpha increases MMP-9 and tenascin expression in bronchial fibroblasts via the transcription factor Ets-1, and suggest a role for Ets-1 in airway remodeling in asthma. PMID- 14734781 TI - Mycobacterium tuberculosis recall antigens suppress HIV-1 replication in anergic donor cells via CD8+ T cell expansion and increased IL-10 levels. AB - Mycobacterium tuberculosis (MTb) is the leading cause of death in the setting of AIDS. MTb enhances the pathogenicity and accelerates the course of HIV disease and, furthermore, infection with HIV-1 increases the risk of reactivation or reinfection with MTb. In this study, we show that host-specific recall responses to one pathogen, MTb, has a direct effect upon the regulation of a second pathogen, HIV-1. Using cells from immunocompetent former tuberculosis (TB) patients who displayed either a persistently positive (responsive) or negative (anergic), delayed-type hypersensitivity (DTH) reaction to intradermal injection of purified protein derivative (PPD), we investigated the effect of recall Ags to MTb upon the replication of HIV-1 primary isolates in vitro. We show that HIV-1 replication of a T cell-tropic isolate was significantly impaired in MTb stimulated PBMC from PPD-anergic donors. Furthermore, these donors displayed a significant increase in CD8(+) T cells and IL-10 levels and lower levels of IL-2 and TNF-alpha relative to PPD-responsive donors in response to PPD stimulation. Strikingly, CD8(+) T cell depletion and blocking of IL-10 significantly increased HIV-1 replication in these PPD-anergic donors, indicating that an immunosuppressive response to MTb recall Ags inhibits HIV-1 replication in PPD anergic individuals. Therefore, immunotherapeutic approaches aimed at recapitulating Ag-specific MTb anergy in vivo could result in novel and effective approaches to inhibit HIV-1 disease progression in MTb/HIV-1 coinfection. PMID- 14734782 TI - Molecular analysis of TCR clonotypes in LGL: a clonal model for polyclonal responses. AB - Large granular lymphocytic (LGL) leukemia is a clonal lymphoproliferative disorder of CTL associated with cytopenias resulting from an immune and cytokine attack on hemopoietic progenitor cells. Extreme clonality of CTL expansions seen in LGL leukemia makes it an ideal model to study the role of the T cell repertoire in other less-polarized immune-mediated disorders. Complementarity determining region 3 (CDR3) of the TCR is a unique Ag-specific region that can serve as a molecular marker, or clonotype, of the disease-specific T cells. We studied the variable portion of the beta-chain spectrum in a cohort of LGL leukemia patients. The CDR3 sequences were determined for the immunodominant clones and used to design clonotype-specific primers. By direct and semi-nested amplification, clonotype amplicons were found to be shared by multiple patients and controls. Analysis of the generated sequences demonstrated that the original clonotypes are rarely encountered in normal control samples; however, high levels of homology were found in both controls and patients. Clonotypes derived from individual LGL patients can be used as tumor markers for the malignant clone. More generally, clonotypic analysis and comparison of the variable portion of the beta-chain CDR3-specific sequences from a large number of patients may lead to better subclassification of not only LGL but also other immune-mediated disorders. PMID- 14734784 TI - Epithelial barriers, compartmentation, and cancer. AB - Epithelial cells, and the tight junctions between them, form a polarized barrier between luminal and serosal fluid compartments and segregate luminal growth factors from their basal-lateral receptors. Breakdown of this barrier should allow access of growth factors in the luminal fluid to their receptors on the basal-lateral cell membranes, as recently demonstrated for heregulin and erbB receptors in airway epithelia. It should also allow luminal growth factors to access the stroma. This property may have adaptive value for epithelial tissues in general, as an elegant response to injury, but may also promote cancer formation in premalignant epithelial tissues in which the tight junctions have become chronically leaky to growth factors. PMID- 14734785 TI - A new look at an old problem: the survival and organ-specific growth of metastases. AB - Despite improvements in cancer detection and therapy, metastatic disease is largely incurable. Recent research indicates that tumor cells disseminate widely early in the process of pathogenesis, and that the survival and proliferation of these cells--and thus the development of metastases--depend on interactions between the disseminated cells and their particular microenvironment. Proliferative signals and the inhibition of proapoptotic responses are both critically involved in the development of clinically significant metastases. Identification of the underlying signaling cascades may provide additional targets for antimetastatic therapy. PMID- 14734786 TI - G proteins in cancer: the prostate cancer paradigm. AB - Signal transduction research investigating mechanisms of androgen-independent prostate cancer cell proliferation has historically focused on the role of androgen and peptide growth factor receptors. More recent work has raised the idea that intracellular signaling mechanisms triggered by extracellular hormonal factors acting through heterotrimeric guanine nucleotide-binding protein (G protein)-coupled receptors (GPCRs) can also mediate and sustain this pathologic process. Prostate cancer patients with advanced disease express elevated levels of GPCRs and GPCR ligands, suggesting that the GPCR system is activated in the cancerous gland and may contribute to tumor growth. Importantly, inhibition of G protein signaling attenuates prostate cancer cell growth in animal models. The nature of intracellular signaling pathways mediating mitogenic effects of GPCRs in prostate cancer is poorly defined, although the G protein-dependent activation of the Ras-to-mitogen-activated protein kinase pathway has emerged as a critical regulatory event. Activated GPCRs may also exert their mitogenic effects in the prostate by activating the androgen receptor. PMID- 14734787 TI - Blunted coronary flow reserve in systemic sclerosis. AB - OBJECTIVES: We investigated whether the non-invasive determination of coronary flow reserve (CFR), as evaluated by transthoracic Doppler echocardiography, might be a potential method to detect early dysfunction of cardiovascular system in patients affected by systemic sclerosis (SSc) without clinical signs or symptoms of cardiac impairment. The possible correlations between the CFR values and the duration of the disease, specific autoantibodies and cutaneous involvement subsets were investigated. METHODS: Forty-four consecutive patients affected by SSc were analysed. The CFR was detected in the distal left anterior descending coronary artery by contrast-enhanced transthoracic second harmonic Doppler in all SSc patients and in 16 healthy controls. CFR was assessed at rest and during hyperaemia induced by administration of adenosine at 0.14 mg/kg/min over 5 min. The CFR was calculated as the ratio between hyperaemic (peak adenosine infusion) and resting peak diastolic velocity (PdvCFR) and resting velocity time integral (VtiCFR). Past medical history was carefully investigated. RESULTS: Both PdvCFR and VtiCFR were significantly reduced in SSc patients when compared with controls (P<0.0001). In particular, both PdvCFR and VtiCFR were significantly lower in patients with dSSc when compared with patients affected by lSSc (P<0.02 and P<0.04 respectively). No statistically significant correlation was found between CFR values and history of smoking, serum levels of cholesterol or triglycerides, blood pressure, age of patients, duration of SSc and serum autoantibody positivity for ANA, ACA and Scl70. CONCLUSIONS: CFR is often reduced in SSc patients. CFR was lower in patients with dSSc than in those affected by lSSc. A reduced CFR value should be considered an indirect sign of heart involvement in scleroderma, but its clinical and prognostic implications need to be clarified. PMID- 14734788 TI - Cytokines regulate fibroblast-like synovial cell differentiation to adipocyte like cells. AB - OBJECTIVES: Our recent work showed that fibroblast-like synovial cells (FLS) could differentiate into adipocyte-like cells in vitro in response to stimulation with peroxisome proliferator-activated receptor gamma (PPAR gamma) ligand. The aim of the present study was to determine the role of cytokines in the regulation of FLS differentiation to adipocyte-like cells. METHODS: FLS isolated from patients with rheumatoid arthritis (RA) and osteoarthritis (OA) and from normal synovial tissues were incubated with the synthetic PPAR gamma ligand troglitazone to induce adipocyte-like differentiation of the cells. RESULTS: Production of interleukin (IL)-6, IL-8 and matrix metalloproteinase-3 was reduced in adipocyte like cells compared with FLS. DNA binding activity of nuclear factor kappa B (NF kappa B) was clearly inhibited in adipocyte-like cells. Cultivation of FLS with interferon gamma (IFN-gamma), tumour necrosis factor-alpha (TNF-alpha) or IL-1 beta inhibited the expression of PPAR gamma as well as CCAAT/enhancer binding protein (C/EBP) nuclear activity, and thus suppressed adipocyte-like cell differentiation in vitro. CONCLUSION: Our results indicate the importance of PPAR gamma and C/EBP in adipocyte-like cell differentiation of FLS and that the process is influenced by inflammatory cytokines, and suggest that the proinflammatory character of FLS in patients with RA is diminished during adipocyte-like cell differentiation. PMID- 14734789 TI - Improved clinical status in fibromyalgia patients treated with individualized homeopathic remedies versus placebo. AB - OBJECTIVE: To assess the efficacy of individualized classical homeopathy in the treatment of fibromyalgia. METHODS: This study was a double-blind, randomized, parallel-group, placebo-controlled trial of homeopathy. Community-recruited persons (N = 62) with physician-confirmed fibromyalgia (mean age 49 yr, s.d. 10 yr, 94% women) were treated in a homeopathic private practice setting. Participants were randomized to receive oral daily liquid LM (1/50,000) potencies with an individually chosen homeopathic remedy or an indistinguishable placebo. Homeopathic visits involved joint interviews and concurrence on remedy selection by two experienced homeopaths, at baseline, 2 months and 4 months (prior to a subsequent optional crossover phase of the study which is reported elsewhere). Tender point count and tender point pain on examination by a medical assessor uninvolved in providing care, self-rating scales on fibromyalgia-related quality of life, pain, mood and global health at baseline and 3 months, were the primary clinical outcome measures for this report. RESULTS: Fifty-three people completed the treatment protocol. Participants on active treatment showed significantly greater improvements in tender point count and tender point pain, quality of life, global health and a trend toward less depression compared with those on placebo. CONCLUSIONS: This study replicates and extends a previous 1-month placebo-controlled crossover study in fibromyalgia that pre-screened for only one homeopathic remedy. Using a broad selection of remedies and the flexible LM dose (1/50,000 dilution factor) series, the present study demonstrated that individualized homeopathy is significantly better than placebo in lessening tender point pain and improving the quality of life and global health of persons with fibromyalgia. PMID- 14734790 TI - Pulmonary involvement in childhood-onset systemic lupus erythematosus: a report of five cases. AB - OBJECTIVE: Systemic lupus erythematosus (SLE) is a chronic systemic disease, which can involve multiple organs such as kidney, skin and brain. Lung is another organ that can be affected. A number of pulmonary complications including pleuritis, pneumonitis, infectious pneumonia, pulmonary haemorrhage, pulmonary hypertension and pneumothorax have been reported in patients with SLE. Pulmonary involvement is relatively frequent in adult patients; it has infrequently been reported in children with SLE. However, pulmonary manifestations may be an initial and/or life-threatening complication of SLE in children. In this paper we aim to emphasize the pulmonary involvement in childhood-onset SLE via description of our patients. METHODS: The patients, who were diagnosed with SLE at the Children's Hospital of Ankara University Medical School between 1993 and 2002, were retrospectively evaluated for evidence of pulmonary involvement. All patients fulfilled at least four of the classification criteria of the American Rheumatism Association. Using a standardized form, we obtained data regarding the age, sex and presenting complaints of the patients, previous therapies given, clinical and laboratory features, treatment and outcome. Informed consent was obtained from all patients. RESULTS: During the 10-yr study period, 16 patients were diagnosed with childhood-onset SLE. Five of them (31%) had pulmonary involvement including acute lupus pneumonitis, invasive pulmonary aspergillosis, cytomegalovirus pneumonia and pulmonary haemorrhage (in two patients). These 5 patients with lupus lung disease are presented in more detail. PMID- 14734791 TI - Leukemia cells and the cytokine network: therapeutic prospects. AB - The network and balance of cytokines is of major importance in maintaining proper homeostasis of hematopoiesis. Abnormalities in this network may result in a variety of blood disorders; however, the role of this network is not clear in leukemia. The use of antineoplastic agents has improved the survival rate of some types of leukemia, and adjunctive therapy with cytokines may be helpful. Chemotherapeutic approaches are no longer the best choice because cytotoxicity may affect normal and leukemic cells, and leukemic cells may develop resistance to the chemotherapeutic agent. Induction of differentiation to a mature phenotype and the control of apoptotic-gene expression have provided other possible alternative therapies. Combined effects of cytokines and vitamin derivatives such as retinoic acid (RA) and 1,25 dihydroxyvitamin D3 (VD3) were found more beneficial than any of these agents individually. These agents exhibit cooperative effects, potentiate each other's effects, or both. Therefore, understanding the hematopoietic actions of these agents, their interactions with their receptors, and their differentiation signaling pathways may result in the design of new therapies. However, the role of cytokines in apoptosis is controversial because in some cases they were found to increase tumor cell resistance to apoptosis-inducing agents. Recent studies in the molecular biology of gene regulation, transcription factors, and repressors have led to new possible approaches such as differentiation therapy for the treatment of leukemia. In addition, the development of drugs that act on the molecular level such as imatinib is just the beginning of a new era in molecular targeted therapy in which the drug acts specifically on the leukemic cell. There are many possible combinations of cytokines, retinoids, and VD3, and perhaps the best therapeutic combination is yet to be described. This minireview is an update on the role of cytokines and the therapeutic potential of combinations with agents such as RA, VD3, and other chemotherapeutic agents. PMID- 14734792 TI - Plasminogen activator inhibitor-1 in the pathogenesis of asthma. AB - Plasminogen activator inhibitor (PAI)-1 is the main inhibitor of the fibrinolytic system and is known to play an essential role in tissue remodeling. Recent evidence indicates that chronic asthma may lead to tissue remodeling such as subepithelial fibrosis and extracellular matrix (ECM) deposition in the airways. However, the role of PAI-1 in asthma is unknown. Recently the mast cell (MC), which plays a major role in asthma, was found as a novel source of PAI-1, and a large number of MCs expressing PAI-1 are infiltrated in the airways of patients with severe asthma. Furthermore, PAI-1-deficient mice show reduced ECM deposition in the airways of a murine model of chronic asthma by inhibiting MMP-9 activity and fibrinolysis. In a human study, the 4G allele frequency was significantly higher in the asthmatic patients than in the control group. In view of the findings that the 4G allele is associated with elevated plasma PAI-1 level, elevated PAI-1 level in the lung may contribute to the development of asthma. In summary, PAI-1 may play an important role in the pathogenesis of asthma and further studies evaluating the mechanisms of PAI-1 action may lead to the development of a novel therapeutic target for the treatment and prevention of asthma. PMID- 14734793 TI - Mammary gland morphogenesis is enhanced by exposure to flaxseed or its major lignan during suckling in rats. AB - The exposure of rats to 10% flaxseed (FS) or an equivalent level of its major lignan, secoisolariciresinol diglucoside (SDG), during suckling enhances mammary gland differentiation, which protects against mammary carcinogenesis at adulthood. We determined whether this diet-induced mammary gland differentiation is mediated through the estrogenic pathway via epidermal growth factor receptor (EGFR) and estrogen receptor (ER) signaling. Rats were fed the AIN-93G basal diet (BD) from day 7 of pregnancy until delivery and then randomized to consume BD, FS, or SDG during lactation. After weaning, female offspring were fed BD throughout the experiment. At postnatal day (PND) 21 and the proestrus phase on PND 49-51, mammary glands of offspring were analyzed for morphology, cell proliferation, and expression of EGFR, epidermal growth factor (EGF), transforming growth factor-alpha, ER-alpha, and ER-beta. At PND 21, compared with the BD control, the number of terminal end buds (TEBs) and terminal ducts were increased by FS, whereas mammary epithelial cell proliferation was increased by both FS and SDG, suggesting that mammary morphogenesis was enhanced. Epithelial EGFR and stromal fibroblast EGF were increased by SDG, whereas epithelial ER-beta was decreased by FS. Conversely, at PND 49-51, a lower number of TEBs but a higher ratio of lobules to TEBs with decreased expression of EGFR or EGF was observed in both treatment groups. EGFR expression was positively associated with EGF expression and cell proliferation in TEB epithelium at PND 21. Urinary lignans of lactating dams were related to their offspring's indices of mammary gland development. In conclusion, exposure to FS or SDG during suckling enhanced mammary gland morphogenesis by modulation of EGFR and ER signaling, which led to more differentiated mammary glands at PND 49-51. The physiological outcomes of FS and SDG were similar, which suggests that SDG is partly responsible for the mammary gland differentiation effect. PMID- 14734794 TI - Sex differences in choline-deficient diet-induced steatohepatitis in mice. AB - Alcoholic steatohepatitis (ASH) and nonalcoholic steatohepatitis (NASH) are common liver diseases in the United States. ASH and NASH occur more frequently in women than in men, and liver injury is also more severe in women. The role of estrogens in ASH has been well established, but their role in NASH has received relatively little study. The purpose of this study was to evaluate the effect of estrogens in methionine-choline deficient diet (MCDD)-induced steatohepatitis in mice. The degree of steatohepatitis was evaluated in males and in intact and ovariectomized females that were fed MCDD for 4 weeks, and in females that were fed MCDD containing tamoxifen. Hematoxylin and eosin-stained sections of livers showed marked steatohepatitis in all experimental groups. Compared to the control group, markers of hepatocyte injury such as aspartate transaminase (AST), alanine transaminase (ALT), and liver triglyceride levels increased significantly in males and in intact and ovariectomized female mice that were fed MCDD. Also, it was interesting that levels of AST and ALT increased much more in the MCDD + tamoxifen group than in the MCDD group. In female mice fed MCDD, hepatocyte proliferative and apoptotic indices increased slightly compared to mice that were fed a normal diet. Based on these results, it can be concluded that MCDD-induced steatohepatitis is comparable in male and female mice, and that ovariectomy or antiestrogen treatment had no protective effect in MCDD-induced steatohepatitis. PMID- 14734795 TI - A novel natural inhibitor of extracellular signal-regulated kinases and human breast cancer cell growth. AB - Water-soluble extracts of edible Vernonia amygdalina leaves were recently reported as potent inhibitors of cultured MCF-7 cells. The mechanism by which V. amygdalina inhibits MCF-7 cell growth has not been previously studied. The objective of this study was to evaluate the effects of V. amygdalina on the activities, DNA synthesis, and subsequent cell growth of extracellular signal regulated protein kinases 1 and 2 (ERKs 1/2;). Treatment of cells with various concentrations (3-100 mg/ml) of water-soluble V. amygdalina extract potently inhibited ERK activities, DNA synthesis (P < 0.005), and cell growth (P < 0.01) in a concentration-dependent fashion, both in the absence and presence of serum. The growth rate of cells pretreated with 10 mg/ml V. amygdalina for 48 hrs before transfer to V. amygdalina-free medium was not significantly different (P > 0.05) from untreated cells. These results suggest that V. amygdalina, at least at concentrations up to 10 mg/ml, exhibits cytostatic action to retard the growth of human breast cancer cells. In addition, the ERK signaling pathways may be one or more of the intracellular targets for V. amygdalina antineoplastic actions. PMID- 14734796 TI - Posttraumatic inflammation is a complex response based on the pathological expression of the nervous, immune, and endocrine functional systems. AB - The successive phases that make up both the local and systemic posttraumatic acute inflammatory response could represent the expression of three concatenated pathological or "primitive" functional systems with trophic properties: the nervous, immune, and endocrine ones. The nervous functional system would play an important role in the phenomenon of ischemia-reperfusion, which would be represented by nutrition by diffusion that is either anaerobic (ischemia) or with defective use of oxygen (reperfusion) and, thus, with a limited energy requirement. The immune functional system would be represented by the infiltration of the tissues by inflammatory cells and bacteria, which would become mediators in providing nutrition to the injured tissues. Although the use of oxygen would still be defective, hypermetabolism and fever would occur. In these inflammatory response phases, the lymphatic is the most important circulation. The endocrine functional system would be the most specialized and would have high energy requirements because it would be represented by the blood capillary-mediated nutrition. Highly specialized epithelial cells would already possess a perfected oxidative metabolism. The successive expression of these three functional systems during embryonic development and also during the evolutionary development of our species could explain why the inflammatory response is a ubiquitous mechanism that is common to multiple diseases, because it is an integrator of the ontogeny and phylogeny. PMID- 14734797 TI - Leptin receptor-deficient MMTV-TGF-alpha/Lepr(db)Lepr(db) female mice do not develop oncogene-induced mammary tumors. AB - Being overweight is a risk factor for postmenopausal breast cancer and is associated with an increased incidence and shortened latency of spontaneous and chemically induced mammary tumors in rodents. However, leptin-deficient obese Lep(ob)Lep(ob) female mice have reduced incidences of spontaneous and oncogene induced mammary tumors. Of interest, leptin enhances the proliferation of human breast cancer cell lines in which leptin receptors are expressed, which suggests that leptin signaling plays a role in tumor development. We evaluated oncogene induced mammary tumor development in obese MMTV-TGF-alpha/Lepr(db)Lepr(db) mice that exhibit a defect in OB-Rb, which is considered to be the major signaling isoform of the leptin receptor. Lepr and MMTV-TGF-alpha mice were crossed, and the offspring were genotyped for oncogene expression and the determination of Lepr status. Lean MMTV-TGF-alpha/Lepr(+)Lepr(+) (homozygous) and MMTV-TGF alpha/Lepr(+)Lepr(db) (heterozygous) mice and obese MMTV-TGF alpha/Lepr(db)Lepr(db) mice were monitored until age 104 weeks. Body weights of MMTV-TGF-alpha/ Lepr(db)Lepr(db) mice were significantly heavier than those of the lean groups. No mammary tumors were detected in MMTV-TGF alpha/Lepr(db)Lepr(db) mice, whereas the incidence of mammary tumors in MMTV-TGF alpha/Lepr(+)Lepr(+) and MMTV-TGF-alpha/ Lepr(+)Lepr(db) mice was 69% and 82%, respectively. Examination of mammary tissue whole mounts indicated an absence of duct formation and branching for MMTV-TGF-alpha/Lepr(db)Lepr(db) mice. Both age at mammary tumor detection and tumor burden (tumors/mouse and tumor weights) were similar for the lean genotypes. Serum leptin levels of MMTV-TGF alpha/Lepr(db)Lepr(db) mice were 12-20-fold higher than levels of lean mice. Thus, despite elevated serum leptin levels, leptin receptor-deficient MMTV-TGF alpha/Lepr(db)Lepr(db) mice do not develop mammary tumors. This study provides additional evidence that leptin and its cognate receptor may be involved in mammary tumorigenesis. PMID- 14734798 TI - Characterization of N-terminal interferon tau mutants: P26L affords enhanced activity and lack of toxicity. AB - Interferon (IFN)-tau is a type I IFN that is responsible for the maternal recognition of pregnancy in ruminants. This protein also has classic IFN-like properties, including antiviral, antiproliferative, and immunomodulatory functions. Using IFN-tau as a model, we examined the structural basis for the activity of type I IFNs, focusing on amino acids within helix A and the first section of the AB loop, which have been proposed as a site for receptor interaction. Six amino-acid substitutions were made that replaced a residue in ovine IFN-tau1mod with the corresponding residue in human IFN-alphaA. Receptor binding was enhanced by a P26L mutation and was reduced by a conservative lysine to-histidine substitution at residue 34. Alterations in the antiviral and antiproliferative activities of the IFN-tau mutants were not always correlated, but both functions were maintained or enhanced relative to the wild-type IFN-tau by the proline-to-leucine mutation at residue 26. In contrast, this mutation did not affect the low in vitro cytotoxicity that is characteristic of ovine IFN tau1mod. Thus, the IFN-tau P26L mutant may have potential as an improved IFN based therapeutic. PMID- 14734799 TI - Selenium attenuates lipopolysaccharide-induced oxidative stress responses through modulation of p38 MAPK and NF-kappaB signaling pathways. AB - Lipopolysaccharide (LPS) produces reactive oxygen species (ROS) and nitric oxide (NO) in macrophages. These molecules are involved in inflammation associated with endotoxic shock. Selenium (Se), a biologically essential trace element, modulates the functions of many regulatory proteins involved in signal transduction and affects a variety of cellular activities, including cell growth and survival. We demonstrate that Se attenuated LPS-induced ROS and NO production in murine macrophage cultures in vitro. This Se-decreased production of NO was demonstrated by decreases in both mRNA and protein expression for inducible NO synthase (iNOS). The preventive effects of Se on iNOS were p38 mitogen-activated protein kinase- and nuclear factor-kappaB-dependent. Se specifically blocked the LPS induced activation of p38 but not that of c-jun-N-terminal kinase and extracellular signal-regulated kinase; the p38-specific pathway was confirmed using p38 inhibitor SB 203580. These results suggest that the mechanism by which Se may act as an anti-inflammatory agent and that Se may be considered as a possible preventive intervention for endotoxemia, particularly in Se-deficient locations. However, the efficacy and safety of Se need to be further investigated, because long-term intake > 0.4 mg Se/day in adults can produce adverse effects. PMID- 14734800 TI - Reduction of ischemia and reperfusion-induced myocardial damage by cytochrome P450 inhibitors. AB - Ischemia and reperfusion both contribute to tissue damage after myocardial infarction. Although many drugs have been shown to reduce infarct size when administered before ischemia, few have been shown to be effective when administered at reperfusion. Moreover, although it is generally accepted that a burst of reactive oxygen species (ROS) occurs at the onset of reperfusion and contributes to tissue damage, the source of ROS and the mechanism of injury is unclear. We now report the finding that chloramphenicol administered at reperfusion reduced infarct size by 60% in a Langendorff isolated perfused rat heart model, and that ROS production was also substantially reduced. Chloramphenicol is an inhibitor of mitochondrial protein synthesis and is also an inhibitor of a subset of cytochrome P450 monooxygenases (CYPs). We could not detect any effect on mitochondrial encoded proteins or mitochondrial respiration in chloramphenicol-perfused hearts, and hypothesized that the effect was caused by inhibition of CYPs. We tested additional CYP inhibitors and found that cimetidine and sulfaphenazole, two CYP inhibitors that have no effect on mitochondrial protein synthesis, were also able to reduce creatine kinase release and infarct size in the Langendorff model. We also showed that chloramphenicol reduced infarct size in an open chest rabbit model of regional ischemia. Taken together, these findings implicate CYPs in myocardial ischemia/reperfusion injury. PMID- 14734801 TI - A tautomeric zinc sensor for ratiometric fluorescence imaging: application to nitric oxide-induced release of intracellular zinc. AB - Zinc is an essential metal ion for human growth and development, the disruption of cellular Zn(2+) homeostasis being implicated in several major disorders including Alzheimer's disease, diabetes, and cancer. The molecular mechanisms of Zn(2+) physiology and pathology are insufficiently understood, however, owing in part to the lack of tools for measuring changes in intracellular Zn(2+) concentrations with high spatial and temporal fidelity. To address this critical need, we have synthesized, characterized, and applied an intracellular fluorescent probe for the ratiometric imaging of Zn(2+) based on a tautomeric seminaphthofluorescein platform. Zin-naphthopyr 1 (ZNP1) affords single excitation, dual-emission ratiometric detection of intracellular Zn(2+) through Zn(2+)-controlled switching between fluorescein and naphthofluorescein tautomeric forms. The probe features visible excitation and emission profiles, excellent selectivity responses for Zn(2+) over competing Ca(2+) and Mg(2+) ions at intracellular concentrations, a dissociation constant (K(d)) for Zn(2+) of <1 nM, and an 18-fold increase in fluorescence emission intensity ratio (lambda(624)/lambda(528)) upon zinc binding. We demonstrate the value of the ZNP1 platform for biological applications by imaging changes in intracellular [Zn(2+)] in living mammalian cells. Included is the ratiometric detection of endogenous pools of intracellular Zn(2+) after NO-induced release of Zn(2+) from cellular metalloproteins. We anticipate that ZNP1 and related probes should find utility for interrogating the biology of Zn(2+). PMID- 14734802 TI - A proton-coupled dynamic conformational switch in the HIV-1 dimerization initiation site kissing complex. AB - In HIV type 1 (HIV-1), the dimerization initiation site (DIS) is the sequence primarily responsible for initiating the noncovalent linkage of two homologous strands of genomic RNA during viral assembly. The DIS loop contains an autocomplementary hexanucleotide sequence and forms a symmetric homodimer through a loop-loop kissing interaction. In a structural rearrangement catalyzed by the HIV-1 nucleocapsid protein (NCp7) and suggested to be associated with maturation of the budded viral particle, the DIS converts from a metastable kissing dimer to an extended duplex. Here, we demonstrate that the DIS kissing dimer displays localized conformational dynamics that result from the specific protonation of the N1 base nitrogen of the DIS loop residue A272 at near-physiological pH. The rate of NCp7-catalyzed maturation of the DIS kissing dimer is also shown to directly correlate with the observed proton-coupled conformational dynamics, where NCp7 is found to convert the dynamic A272 protonated state with a faster rate. Taken together, these results reveal a previously undescribed role for base protonation in modulating local RNA structure and demonstrate a mechanism for promoting the chaperone-mediated structural rearrangement of a kinetically trapped RNA conformational state. PMID- 14734803 TI - Complex nested promoters control tissue-specific expression of acetyl-CoA carboxylase genes in wheat. AB - Cis-acting regulatory elements of the wheat acetyl-CoA carboxylase (ACC) gene family were identified by comparing the promoter activity of 5' end gene fragments fused to a reporter gene in two transient expression systems: wheat protoplasts and epidermal cells of mature embryos. Expression of the plastid and the cytosolic ACC genes is each driven by two nested promoters responsible for the synthesis of two transcript types. The internal promoter is located in an intron removed from transcripts originating at the first promoter. These complex promoters, which are different for the cytosolic and plastid ACC genes, control tissue-specific expression of the enzymatic activity supplying cytosolic, plastid, and mitochondrial pools of malonyl-CoA. The activity of one such complex promoter, driving expression of one of the cytosolic ACC genes, was studied throughout development of transgenic wheat plants carrying a full-length promoter reporter gene fusion. High activity of the promoter was detected in the coleoptile, in the upper sheath section of the leaf, on the top surface of the ovary, in some sections of the main veins in the lemma and glume, and in abaxial epidermis hair cells of the lemma, glume, and rachis. The findings are consistent with the developmental and environmental requirements for very-long-chain fatty acids and flavonoids, whose synthesis begins with the ACC reaction in the cytosol of these specific cell types. PMID- 14734804 TI - Antibody to DNA detects scrapie but not normal prion protein. AB - Prion diseases, a group of fatal neurodegenerative disorders, are characterized by the presence of the abnormal scrapie isoform of prion protein (PrP(Sc)) in affected brains. A conformational change is believed to convert the normal cellular prion protein into PrP(Sc). Detection of PrP(Sc) for diagnosis and prophylaxis is impaired because available Abs recognizing epitopes on PrP fail to distinguish between PrP(Sc) and normal cellular prion protein. Here, we report that an anti-DNA Ab, OCD4, as well as gene 5 protein, a well established DNA binding protein, capture PrP from brains affected by prion diseases in both humans and animals but not from unaffected controls. OCD4 appears to immunoreact with DNA (or a DNA-associated molecule) that forms a conformation-dependent complex with PrP in prion diseases. Whereas PrP immunocaptured by OCD4 is largely protease-resistant, a fraction of it remains protease-sensitive. Moreover, OCD4 detects disease-associated PrP >10 times more efficiently than a widely used Ab to PrP. Our finding that anti-DNA Abs and gene 5 protein specifically target disease-associated DNA-PrP complexes in a wide variety of species and disease phenotypes opens new avenues in the study and diagnosis of prion diseases. PMID- 14734805 TI - DNA-PKcs function regulated specifically by protein phosphatase 5. AB - Unrepaired DNA double-strand breaks can lead to apoptosis or tumorigenesis. In mammals double-strand breaks are repaired mainly by nonhomologous end-joining mediated by the DNA-PK complex. The core protein of this complex, DNA-PKcs, is a DNA-dependent serine/threonine kinase that phosphorylates protein targets as well as itself. Although the (auto)phosphorylation activity has been shown to be essential for repair of both random double-strand breaks and induced breaks at the immunoglobulin locus, the corresponding phosphatase has been elusive. In fact, to date, none of the putative phosphatases in DNA double-strand break repair has been identified. Here we show that protein phosphatase 5 interacts with DNA-PKcs and dephosphorylates with surprising specificity at least two functional sites. Cells with either hypo- or hyperphosphorylation of DNA-PKcs at these sites show increased radiation sensitivity. PMID- 14734806 TI - Histone deacetylase (HDAC) inhibitor activation of p21WAF1 involves changes in promoter-associated proteins, including HDAC1. AB - Histone deacetylase (HDAC) inhibitors (HDACi) cause cancer cell growth arrest and/or apoptosis in vivo and in vitro. The HDACi suberoylanilide hydroxamic acid (SAHA) is in phase I/II clinical trials showing significant anticancer activity. Despite wide distribution of HDACs in chromatin, SAHA alters the expression of few genes in transformed cells. p21(WAF1) is one of the most commonly induced. SAHA does not alter the expression of p27(KIPI), an actively transcribed gene, or globin, a silent gene, in ARP-1 cells. Here we studied SAHA-induced changes in the p21(WAF1) promoter of ARP-1 cells to better understand the mechanism of HDACi gene activation. Within 1 h, SAHA caused modifications in acetylation and methylation of core histones and increased DNase I sensitivity and restriction enzyme accessibility in the p21(WAF1) promoter. These changes did not occur in the p27(KIPI) or epsilon-globin gene-related histones. The HDACi caused a marked decrease in HDAC1 and Myc and an increase in RNA polymerase II in proteins bound to the p21(WAF1) promoter. Thus, this study identifies effects of SAHA on p21(WAF1)-associated proteins that explain, at least in part, the selective effect of HDACi in altering gene expression. PMID- 14734807 TI - Molecular dissection of water and glycerol permeability of the aquaglyceroporin from Plasmodium falciparum by mutational analysis. AB - The selectivity of aquaporins for water and solutes is determined by pore diameter. Paradoxically, the wider pores of glycerol facilitators restrict water passage by an unknown mechanism. Earlier we characterized an aquaglyceroporin from Plasmodium falciparum with high permeability for both glycerol and water. We use point mutations to demonstrate that amino acids directly lining the pore are not responsible for the excellent water permeability of the Plasmodium aquaglyceroporin but affect permeability of pentitols. Within a conserved WET triad in the extracellular C-loop we identified a Plasmodium aquaglyceroporin specific glutamate (E125) located in proximity to a conserved arginine (R196) at the pore mouth. Mutation of E125 to serine largely abolished water permeability. Concomitantly, the activation energy for water permeation was increased by 4 kcal/mol. Mutation of the adjacent tryptophan to cysteine led to irreversible inhibition of water passage by Hg(2+). This unequivocally proves the proximity of the couple W124/E125 close to the pore mouth. We conclude that in the Plasmodium aquaglyceroporin the electrostatic environment at the extracellular pore entry regulates water permeability. PMID- 14734808 TI - Complement regulation at the molecular level: the structure of decay-accelerating factor. AB - The human complement regulator CD55 is a key molecule protecting self-cells from complement-mediated lysis. X-ray diffraction and analytical ultracentrifugation data reveal a rod-like arrangement of four short consensus repeat (SCR) domains in both the crystal and solution. The stalk linking the four SCR domains to the glycosylphosphatidylinositol anchor is extended by the addition of 11 highly charged O-glycans and positions the domains an estimated 177 A above the membrane. Mutation mapping and hydrophobic potential analysis suggest that the interaction with the convertase, and thus complement regulation, depends on the burial of a hydrophobic patch centered on the linker between SCR domains 2 and 3. PMID- 14734809 TI - Diffusion at finite speed and random walks. AB - Diffusion, which occurs with infinite speed, results from a random walk with steps of finite speed. We resolve this paradox and derive a modified diffusion equation with finite speed. PMID- 14734810 TI - An analysis of the gene expression program of mammalian neural progenitor cells. AB - A diverse range of neural cell types is generated from a pool of dividing stem and progenitor cells in an orderly manner during development. Little is known of the molecular and cellular biology underpinning the intrinsic control of this process. We have used a nonbiased method to purify populations of neural progenitor cells from the murine CNS to characterize the gene expression program of mammalian retinal progenitor cells. Analysis of these data led to the identification of a core set of >800 transcripts enriched in retinal progenitor cells compared to both their immediate postmitotic progeny and to differentiated neurons. This core set was found to be shared by progenitors in other regions of the developing CNS, with important regional differences in key functional families. In addition to providing an expression fingerprint of this cell type, this set highlights several key aspects of progenitor biology. PMID- 14734811 TI - A genomewide oscillation in transcription gates DNA replication and cell cycle. AB - Microarray analysis from a yeast continuous synchrony culture system shows a genomewide oscillation in transcription. Maximums in transcript levels occur at three nearly equally spaced intervals in this approximately 40-min cycle of respiration and reduction. Two temporal clusters (4,679 of 5,329) are maximally expressed during the reductive phase of the cycle, whereas a third cluster (650) is maximally expressed during the respiratory phase. Transcription is organized functionally into redox-state superclusters with genes known to be important in respiration or reduction being synthesized in opposite phases of the cycle. The transcriptional cycle gates synchronous bursts in DNA replication in a constant fraction of the population at 40-min intervals. Restriction of DNA synthesis to the reductive phase of the cycle may be an evolutionarily important mechanism for reducing oxidative damage to DNA during replication. PMID- 14734812 TI - Persistently active cannabinoid receptors mute a subpopulation of hippocampal interneurons. AB - Cortical information processing requires an orchestrated interaction between a large number of pyramidal cells and albeit fewer, but highly diverse GABAergic interneurons (INs). The diversity of INs is thought to reflect functional and structural specializations evolved to control distinct network operations. Consequently, specific cortical functions may be selectively modified by altering the input-output relationship of unique IN populations. Here, we report that persistently active cannabinoid receptors, the site of action of endocannabinoids, and the psychostimulants marijuana and hashish, switch off the output (mute) of a unique class of hippocampal INs. In paired recordings between cholecystokinin-immunopositive, mossy fiber-associated INs, and their target CA3 pyramidal cells, no postsynaptic currents could be evoked with single presynaptic action potentials or with repetitive stimulations at frequencies <25 Hz. Cannabinoid receptor antagonists converted these "mute" synapses into high fidelity ones. The selective muting of specific GABAergic INs, achieved by persistent presynaptic cannabinoid receptor activation, provides a state dependent switch in cortical networks. PMID- 14734813 TI - Inhibition of kinesin motility by ADP and phosphate supports a hand-over-hand mechanism. AB - The motor protein kinesin couples a temporally periodic chemical cycle (the hydrolysis of ATP) to a spatially periodic mechanical cycle (movement along a microtubule). To distinguish between different models of such chemical-to mechanical coupling, we measured the speed of movement of conventional kinesin along microtubules in in vitro motility assays over a wide range of substrate (ATP) and product (ADP and inorganic phosphate) concentrations. In the presence and absence of products, the dependence of speed on [ATP] was well described by the Michaelis-Menten equation. In the absence of products, the K(M) (the [ATP] required for half-maximal speed) was 28 +/- 1 microM, and the maximum speed was 904 nm/s. P(i) behaved as a competitive inhibitor with K(I) = 9 +/- 1 mM. ADP behaved approximately as a competitive inhibitor with K(I) = 35 +/- 2 microM. The data were compared to four-state kinetic models in which changes in nucleotide state are coupled to chemical and/or mechanical changes. We found that the deviation from competitive inhibition by ADP was inconsistent with models in which P(i) is released before ADP. This is surprising because all known ATPases (and GTPases) with high structural similarity to the motor domains of kinesin release P(i) before ADP (or GDP). Our result is therefore inconsistent with models, such as one-headed and inchworm mechanisms, in which the hydrolysis cycle takes place on one head only. However, it is simply explained by hand-over-hand models in which ADP release from one head precedes P(i) release from the other. PMID- 14734814 TI - Control of calcium oscillations by membrane fluxes. AB - It is known that Ca(2+) influx plays an important role in the modulation of inositol trisphosphate-generated Ca(2+) oscillations, but controversy over the mechanisms underlying these effects exists. In addition, the effects of blocking membrane transport or reducing Ca(2+) entry vary from one cell type to another; in some cell types oscillations persist in the absence of Ca(2+) entry (although their frequency is affected), whereas in other cell types oscillations depend on Ca(2+) entry. We present theoretical and experimental evidence that membrane transport can control oscillations by controlling the total amount of Ca(2+) in the cell (the Ca(2+) load). Our model predicts that the cell can be balanced at a point where small changes in the Ca(2+) load can move the cell into or out of oscillatory regions, resulting in the appearance or disappearance of oscillations. Our theoretical predictions are verified by experimental results from HEK293 cells. We predict that the role of Ca(2+) influx during an oscillation is to replenish the Ca(2+) load of the cell. Despite this prediction, even during the peak of an oscillation the cell or the endoplasmic reticulum may not be measurably depleted of Ca(2+). PMID- 14734815 TI - RNase H2 of Saccharomyces cerevisiae is a complex of three proteins. AB - The composition of RNase H2 has been a long-standing problem. Whereas bacterial and archaeal RNases H2 are active as single polypeptides, the Saccharomyces cerevisiae homolog, Rnh2Ap, when expressed in Escherichia coli, fails to produce an active RNase H2. By affinity chromatography purification and identification of polypeptides associated with a tagged S.cerevisiae Rnh2Ap, we obtained a complex of three proteins [Rnh2Ap (Rnh201p), Ydr279p (Rnh202p) and Ylr154p (Rnh203p)] that together are necessary and sufficient for RNase H2 activity [correction]. Deletion of the gene encoding any one of the proteins or mutations in the catalytic site in Rnh2A led to loss of RNase H2 activity. Even when S.cerevisiae RNase H2 is catalytically compromised, it still exhibits a preference for cleavage of the phosphodiester bond on the 5' side of a ribonucleotide deoxyribonucleotide sequence in substrates mimicking RNA-primed Okazaki fragments or a single ribonucleotide embedded in a duplex DNA. Interestingly, Ydr279p and Ylr154p have homologous proteins only in closely related species. The multisubunit nature of S.cerevisiae RNase H2 may be important both for structural purposes and to provide a means of interacting with other proteins involved in DNA replication/repair and transcription. PMID- 14734816 TI - Highly sensitive amperometric detection of genomic DNA in animal tissues. AB - A simple and highly sensitive method for the detection of genomic DNA in tissue samples is described. It is based on amperometric detection of target DNA by forming an analyte/polymeric activator bilayer on a gold electrode. The biotinylated target DNA is hybridized to oligonucleotide capture probes immobilized on the gold electrode, forming the first layer. A subsequent binding of glucose oxidase-avidin conjugate to the target DNA and the introduction of a second layer of a redox polymer to the electrode, via layer-by-layer electrostatic self-assembly, allow for electrochemical detection of the catalytic oxidation current of glucose in a PBS solution. Less than 2.0 fg of rat genomic DNA, for both regulated and house-keeping genes, can be easily detected in 2.5 microl droplets. The proposed procedure shows very high specificity for genomic DNA in a RT-PCR mixture. PMID- 14734817 TI - Rapid and accurate characterisation of short tandem repeats by MALDI-TOF analysis of endonuclease cleaved RNA transcripts. AB - We describe the application of matrix-assisted laser desorption/ionisation time of-flight mass spectrometry (MALDI-TOF MS) for the characterisation of short tandem repeat (STR) sequences by the analysis of endonuclease cleaved RNA transcripts. Several simple bovine STR loci as well as interrupted and compound microsatellites were chosen as model loci to evaluate the capabilities of MALDI TOF MS for STR analysis. In short, the described approach consists of a PCR amplification of the investigated STR sequence, which then is transcribed into RNA and cleaved by G-specific RNase T1. Base-specific cleavage of the transcript results in high informative fragment patterns from both the repetitive core sequence and the flanking region. Since sequence specificity from endonuclease cleavage is combined with the accuracy of MALDI-TOF measurements, this technique allows for fast and reliable determination of simple repeat lengths as well as for further characterisation of STR allele sequences, which is of high interest especially in more complex STR loci. PMID- 14734818 TI - Establishment of stable melanoma cell line expressing a novel gene, jpk, using a tetracycline-controlled gene expression system. AB - Jpk, originally isolated as an associating factor with the position-specific regulatory element of Hoxa-7, was found to be toxic to Escherichia coli (1) and to F9 teratocarcinoma cells (2) when transiently transfected and expressed. To investigate the possibility of tumor gene therapy using Jpk, its effect was tested in B16F10 murine melanoma cells. Because Jpk reduces the viability of B16F10 cells when transiently expressed, the Jpk gene was cloned into a tetracycline-controlled gene expression vector, pRetro-On to circumvent the lethal effect in unwanted situations. The retroviral plasmid pRetroJpk purified from the packaging cell was infected into B16F10 melanoma cells and screened in the presence of puromycin. Out of a total of 53 stable clones selected with puromycin, two clones overexpressed Jpk at more than twice the level when induced by doxycycline, a tetracycline-derivative, which implies the amount of the Jpk exhibiting the toxicity is critical. Although these clones control only low levels of Jpk, overexpression of the established melanoma cell line may help us decipher the function of Jpk and apply it as a tumor therapeutic gene in the future. PMID- 14734819 TI - Variations in transfection efficiency of VEGF165 and VEGF121-cDNA: its effects on proliferation and migration of human endothelial cells. AB - Little is known about the expression pattern of vascular endothelial growth factor (VEGF) among smooth muscle cells of different arterial regions. Therefore, we have conducted studies aimed at increasing expression of VEGF in cultured human smooth muscle cells (SMCs) from different sites: aorta, umbilical artery, and coronary artery. Two plasmids harboring human VEGF121 and VEGF165 isoforms, respectively, were constructed and lipotransfected into vascular SMCs, using the Fu-GENE 6. Extensive optimization of transfection conditions were performed prior to this. Different basal levels of VEGF were observed between cell types: from 0.51-0.95 pg/mL/micrograms protein in umbilical artery, through 2.32-2.39 pg/mL/micrograms protein in coronary artery, to 5.45-7.52 pg/mL/micrograms protein in aortic SMCs. Significant differences in responses to transfection were also observed: The increase in VEGF production was most pronounced in umbilical artery SMCs (e.g., with 4 micrograms VEGF121-cDNA /in the wells)- an approximate 600-fold as opposed to an 18-fold increase in aortic SMCs and a 29-fold increase in coronary artery SMCs. In addition, we observed significant increases in proliferation rate of aortic and coronary endothelial cells (ECs), after incubation with conditioned medium from VEGF-transfected SMCs. Observed changes differed in relation to cell origin and isoform. PMID- 14734820 TI - Expression of modified Cry1Ac gene of Bacillus thuringiensis in transgenic tobacco plants. AB - Several mutations were introduced into the Cry1Ac toxin gene, resulting in four variants with altered sequences that were responsible for low expression of the toxin in transgenic plants. These variants were as follows: V1, with modified three A/T-rich regions, including the first signal of transcription termination; V2, with modified five putative polyadenylation signals (polyadenylation signals PAS) and the second signal of transcription termination; V3, with four initial AUUUA motifs; V4, with modification of six PASs, four AUUUA motifs, as well as the first and the second signals of transcription termination. The modified variants and the initial WT gene were cloned into the binary vector pBI121 and introduced into tobacco plants (Nicotiana tabacum) by Agrobacterium tumefaciens mediated transformation. The presence of transgenes in the tobacco plants was confirmed by the polymerase chain reaction (PCR) method. The expression of particular variants of the Cry1Ac gene in tobacco was assayed using Western blotting with antibodies against the domain II of the Cry1Ac toxin. The average expression of WT was estimated to be 0.0025% of soluble proteins, and the expression levels of modified variants were 0.004%, 0.0098%, 0.0125%, and 0.0043% for V1, V2, V3, and V4, respectively. In this article we described the construction of a variant of the Cry1Ac gene (V3) with 12 point mutations leading to an average level of expression in transgenic plants five times higher than that observed in the case of the WT gene. Our results have shown for the first time that the modification of AUUUA sequences has a significant effect on the expression of the Cry1Ac gene in transgenic plants. PMID- 14734821 TI - A novel simple and rapid PCR-based site-directed mutagenesis method. AB - Site-directed mutagenesis (SDM) is a powerful tool for exploring protein structure and function, and several procedures adjusted to specific purposes are still being developed. Herein we describe a straightforward and efficient method with versatile applications for introducing site-specific alterations in any deoxyribonucleic acid (DNA) sequence cloned in a plasmidic expression vector. In this polymerase chain reaction (PCR)-based SDM method, forward and reverse primers are used to amplify the plasmid containing the sequence of interest. The primers are designed so that the desired modifications are introduced at the 5' end of one of the primers, whereas the other primer starts with the nucleotide at position (-1) of the one to be modified. The PCR is carried out using Pfu DNA polymerase. The blunt-ended PCR-generated DNA fragment is self-ligated and used to transform Escherichia coli. Mutant clones are screened by colony hybridization using the mutagenic primer as probe and the presence of the mutation is confirmed by direct DNA sequencing. This procedure was used efficiently to introduce substitutions, deletions, and insertions in the DNA sequences coding for a recombinant form (scFv) of antibody 107 specific of the human CR3 molecule, the rat alpha integrin CD11b A-domain and the human CD8beta cloned in pPICZalphaB, pGEX-2T, and CDM8 expression vectors, respectively. PMID- 14734822 TI - Cloning genes from a library using a clustering strategy and PCR. AB - A new polymerase chain reaction (PCR)-based method is described for the isolation of clones of interest from a library when only part of a sequence is available. In actuality, this occurs with many genomes that have been partially sequenced using a random strategy. The method presented here, discriminating clusters by PCR (DCbyPCR), is a nonradioactive and improved alternative to colony hybridization. PMID- 14734825 TI - Method for introducing specific and unmarked mutations into the chromosome of Streptococcus pneumoniae. AB - This work describes a procedure for the generation of site-specific mutations into the chromosome of Streptococcus pneumoniae that does not involve the use of an antibiotic resistance marker. A linear fragment of transforming deoxyribonucleic acid (DNA) is constructed by polymerase chain reaction (PCR) (gene splicing by overlap extension) and used to transform competent cells of S. pneumoniae. Selection of transformants is performed by PCR, and typically, 1% of the transformed cells show the expected mutation. By this protocol it is possible to change a single base pair into the pneumococcal genome, as well as obtaining in-frame deletions and insertions. PMID- 14734824 TI - Specificity and performance of PCR detection assays for microbial pathogens. AB - PCR has become a widely used tool for detection, identification and differentiation of pathogenic microorganisms in diagnosis of animal and human diseases. However, quite a number of currently used protocols can be further optimized to exclude nonspecific reactions. On the one hand, target sequences as defined by primer binding sites should be checked carefully for the absence of significant homologies to other organisms in order to insure high specificity of detection. A major part of PCR assays is still based on target sequences in the ribosomal RNA operon, but, as the differentiating potential of this region is limited, genes encoding cellular proteins, such as toxins, surface antigens or enzymes, have been shown to be a viable alternative in many instances. On the other hand, various approaches are available to improve the performance of the amplification reaction itself. The kinetics of amplification is known to be heavily dependent on primer-to-template ratio, efficiency of primer annealing and enzyme-to-template ratio. In the present paper, recently published PCR detection assays for microorganisms, particularly bacterial pathogens, are reviewed and optimization strategies are explained. The practical implications and epidemiological consequences of routine use of PCR in the diagnostic laboratory are also discussed. PMID- 14734823 TI - Generation and production of engineered antibodies. AB - Various forms of recombinant monoclonal antibodies are being used increasingly, mainly for therapeutic purposes. This review specifically focuses on what is now called antibody engineering, and discusses the generation of chimeric, humanized, and fully human recombinant antibodies, immunoglobulin fragments, and artificial antigen-binding molecules. Since the production of recombinant antibodies is a limiting factor in their availability, and a shortage is expected in the future, different expression systems for recombinant antibodies and transgenic organisms as bioreactors are also discussed, along with their advantages and drawbacks. PMID- 14734826 TI - Rapid genotyping of the osteoporosis-associated polymorphic transcription factor Sp1 binding site in the COL1A1 gene by pyrosequencing. AB - We describe a novel polymerase chain reaction (PCR) and deoxyribonucleic acid (DNA) sequencingbased assay for rapid genotyping of the polymorphic Sp1 binding site in the COL1A1 gene (1). A single nucleotide G-->T substitution polymorphism at this GC-rich site has recently been reported to be a predictive genetic marker for low bone mineral density (BMD). To simplify screening for this marker, we optimized PCR conditions and subjected the amplicons to pyrosequencing, which is a convenient high-throughput sequence analysis technique, readily amenable to automation. The analysis of 200 deidentified convenience DNA samples extracted from blood revealed genotype frequences in Hardy-Weinberg equilibrium (SS 68.0%, Ss 28.5%, and ss 3.5%) in agreement with other studies of European populations. This study demonstrates for the first time that pyrosequencing can be used for rapid identification of the osteoporosis-associated single nucleotide polymorphism (SNP) in the COL1A1 gene. PMID- 14734827 TI - Increase in cardiovascular pathology in female Sprague-Dawley rats following chronic treatment with 2,3,7,8-tetrachlorodibenzo-p-dioxin and 3,3',4,4',5 pentachlorobiphenyl. AB - The effects of chronic exposure to dioxin (2,3,7,8,-tetrachlorodibenzo-pdioxin [TCDD]) and a dioxin-like compound (3,3',4,4',5-pentachlorobiphenyl [PCB126]) on the cardiovascular system were evaluated in female Harlan Sprague-Dawley rats as part of an ongoing National Toxicology Program investigation. The animals were gavage treated 5 d per week with up to 1000 ng of PCB126 per kilogram of body weight per day or up to 100 ng of TCDD per kilogram of body weight per day for up to 2 yr. The control animals received only a corn oil/acetone vehicle (99:1 mixture). The corresponding stop-study groups received the highest doses for 31 wk and then received only the vehicle for the remainder of the study. After a full necropsy of all animals, a complete set of tissues was examined microscopically. Administration of each compound was associated with treatment related increases in the incidences of degenerative cardiovascular lesions. Cardiomyopathy and chronic active arteritis increased in a dose-related manner in all groups treated with PCB126 or with TCDD. Increased incidences were also observed in the stop-study groups, indicating that a shorter term exposure may produce some effects. The average severity of cardiomyopathy was minimal or slightly greater in all dose groups, including the controls. Chronic active arteritis occurred primarily in the mesentery and pancreas, although the rectum, liver, heart, ovary, uterus, and glandular stomach in the PCB126 study and the liver and ovary in the TCDD study were affected in a few of the dosed animals. The authors' investigations indicate that the rat cardiovascular system is a target for dioxin toxicity, which increases the incidence of spontaneous cardiomyopathy and arteritis. PMID- 14734828 TI - Emetine inhibits glycolysis in isolated, perfused rat hearts. AB - This work was designed to test whether phosphofructokinase is a target for emetine action on the heart. The effects of 37 microM emetine on the activities of phosphofructokinase and hexokinase were measured in homogenates from perfused hearts. The action of increasing concentrations of emetine was determined in nonperfused heart homogenates. The effect of 37 microM emetine or control solutions on the concentration of fructose-6-phosphate and fructose- 1,6 phosphate was measured. The effect of 37 microM emetine or control perfusion on the utilization of fructose-6-phosphate by phosphofructokinase in centrifugation supernatants of homogenates and in reconstituted 27,000g pellets was measured. Double-reciprocal plots of fructose-6-phosphate concentrations vs phosphofructokinase activities were plotted. Emetine decreased phosphofructokinase activity in homogenates from both perfused and nonperfused hearts. Emetine did not inhibit cardiac hexokinase activity. In homogenates from nonperfused hearts, the maximal inhibition with high concentrations of emetine was approx 50%. Emetine perfusion caused a simultaneous increase in the phosphofructokinase substrate fructose-6-phosphate and a decrease in the phosphofructokinase product fructose-1,6-bisphosphate. Phosphofructokinase and, consequently, glycolytic flux appear to be subcellular targets for emetine in the heart. Homogenate centrifugation studies indicate that emetine acts on bound rather than unbound phosphofructokinase. The inhibition may be uncompetitive in nature. PMID- 14734829 TI - Effects of angiotensin-converting enzyme inhibitor on delayed-onset doxorubicin induced cardiotoxicity. AB - Childhood survivors of cancer who are treated with anthacycline chemotherapy, such as doxorubicin (DOX), can develop late-onset cardiomyopathy years after chemotherapy. The mechanism(s) for progression of anthracycline cardiotoxicity to late cardiomyopathy is unknown. Because angiotensin II has been implicated in the progression of other cardiomyopathies, this investigation was undertaken to determine whether treatment with an angiotensinconverting enzyme (ACE) inhibitor, lisinopril, reduces the time-dependent effects of doxorubicin on cardiac gene expression and myocellular apoptosis. A single dose of saline (control) or doxorubicin (DOX treated; 2 mg/kg iv) was administered to rabbits. Control and DOX-treated groups were also subgrouped to receive lisinopril and designated as lisinopril or DOX + lisinopril, respectively (1 mg/kg/d oral), for 10 wk. Histopathology, as determined at the light and ultrastructural level, was consistent with a reduced number of cardiomyocytes relative to interstitial cells in the left ventricle (LV) of the DOX-treated group compared with control and DOX + lisinopril groups. Gene expression of the pro-atrial naturetic peptide (ANP), quantified by steady-state messenger ribonucleic acid (mRNA) levels with reverse transcriptase polymerase chain reaction (RT-PCR) and Southern blotting, increased approx 12-fold (n = 10; p < 0.05) in the LV of DOX-treated groups compared to control and DOX + lisinopril groups. Increased ANP mRNA expression following doxorubicin dosing was localized predominantly in ventricular myocytes by in situ hybridization. Deoxyribonucleic acid (DNA) fragmentation, determined by TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling), was increased in both DOX-treated and DOX + lisinopril groups compared to the control group. Lisinopril prevented both late-onset increased ventricular ANP expression and subsequent DOX-induced myocyte loss. The authors speculate that these protective effects of lisinopril are related to reduced ANP expression and myocyte loss, the latter possibly mediated by effects on myocellular apoptosis. PMID- 14734830 TI - Grape seed proanthocyanidins induce pro-oxidant toxicity in cardiomyocytes. AB - Grape seed proanthocyanidin extract (GSPE), a polyphenolic compound with antioxidant properties, may protect against cardiac ischemia and reperfusion injury. However, its potential toxicity at higher doses is unknown. The authors tested the effects of GSPE on reactive oxygen species (ROS) generation, cell survival, lactate dehydrogenase (LDH) release, and caspase- 3 activity using chick cardiomyocytes incubated with GSPE at 5, 10, 50, 100, or 500 micrograms/mL in medium for 8 h. Exposure to increasing concentrations of GSPE (100 or 500 micrograms/mL) resulted in an increase in ROS generation and cell death as measured by propidium iodide uptake and LDH release. Caspase-3 activity was significantly increased fourfold in cells exposed to GSPE 500 micrograms/ mL compared to controls; this was abolished by the selective caspase-3 inhibitor Ac Asp-Gln-Thr-Asp-H (50 microM), which also significantly reduced the cell death resulting from GSPE (500 micrograms/mL). The antioxidant N-acetylcysteine (NAC, 100 microM) reduced cell death induced by GSPE (500 micrograms/mL) but failed to attenuate caspase-3 activation. Collectively, the authors conclude that higher doses of GSPE could cause apoptotic cell injury via effector caspase-3 activation and subsequent induction of ROS generation. Consumers may take higher doses of dietary supplements in the belief that natural herbs have no major side effects. This study demonstrates that dosages of GSPE should be optimized to avoid potential harmful pro-oxidant effects. PMID- 14734832 TI - Cardiovascular effects of inhaled diesel exhaust in spontaneously hypertensive rats. AB - Particulate matter air pollution is associated with increased cardiovascular mortality. The present study examined the cardiac effects of diesel exhaust exposure in spontaneously hypertensive rats. These rats (4 mo old, n = 6 males and 4-6 females/concentration) were exposed to one of five diesel exhaust levels (0, 30, 100, 300, and 1000 micrograms particles/m3) for 6 h per day for 7 d. Electrocardiographic measurements were obtained by radiotelemetry beginning 3 d prior to exposure and ending 4 d after exposure cessation. Control rats displayed a reduced daytime heart rate from the beginning of the protocol, whereas exposed rats maintained a significantly elevated heart rate throughout the exposure. Daytime heart rate values for male control rats averaged 265 +/- 5 beats/min (mean +/- standard error [SE]), whereas values for exposed rats averaged 290 +/- 7 beats/min. This difference persisted during the evenings of the exposure period but was not observed at any time during the preexposure or postexposure periods. The PQ interval, an index of atrioventricular node sensitivity, was significantly prolonged among exposed animals in a concentration-dependent manner. Increased heart rate with prolongation of the PQ interval may represent a substrate for ventricular arrhythmias. These results concur with previous reports suggesting that realistic exposure concentrations of air pollution affect the pacemaking system of rats. PMID- 14734831 TI - Cardiac toxic effects of trans-2-hexenal are mediated by induction of cardiomyocyte apoptotic pathways. AB - Aldehydes are ubiquitous pollutants with well-indicated but ill-defined cardiovascular toxicity. To investigate the direct toxic effects of environmental aldehyde exposure on the myocardium, 8-wk-old male ICR (Institute of Cancer Research) strain mice were gavage fed trans-2-hexenal (0.1, 1, 10, or 50 mg/kg/wk) or corn oil (vehicle) for 4 wk, during which cardiac function, myocardial morphology, cardiomyocyte apoptosis, and the cytochrome cmediated caspase activation apoptotic pathway were determined. Quantification by enzyme linked immunosorbent assay (ELISA) revealed that aldehyde- protein adducts increase in mouse hearts following hexenal treatment, whereas echocardiographic analysis displayed a significant impairment of basal left-ventricular contractile function. Both histological analysis and TUNEL (terminal deoxynucleotidyl transferase-mediated nick-end labeling) staining indicated condensed nuclei and a significant increase in cardiomyocyte apoptosis in these mice, but immunohistochemistry-based confocal microscope revealed no marked myofibril disarray. Release of cytochrome c from mitochondria into the cytosol, concomitant with activation of caspase-3 and -9, was also found in hexenal-treated groups. In addition, isolated cardiac mitochondria formed hexenal-protein adducts when treated with hexenal, providing indirect evidence that the cardiac mitochondrion is one of primary subcellular targets of aldehyde toxins. These findings suggest that trans-2-hexenal exposure results in direct cardiac toxicity through, at least in part, induction of mitochondrial cytochrome c release-mediated apoptosis in cardiomyocytes, indicating that the cardiac mitochondrion is one of principal subcellular targets of aldehyde toxins. PMID- 14734833 TI - Diabetes enhances lipopolysaccharide-induced cardiac toxicity in the mouse model. AB - Diabetic patients have a higher rate of mortality from sepsis than do their nondiabetic septic counterparts. The hypothesis in this study is that chronic diabetes may make cardiovascular systems more sensitive to septicemia. To test this hypothesis, the authors investigated the effect of diabetes on endotoxin- induced cardiac toxicity. Diabetes was induced in FVB mice by injecting a single dose (150 mg/kg) of streptozotocin. Two months after streptozotocin treatment, the diabetic mice were treated with lipopolysaccharide by intraperitoneal injection at 2 mg/kg. Cardiac toxicity was evaluated by measuring levels of serum cardiac enzymes and cardiac morphology at 1 h, 4.5 h, and 24 h after lipopolysaccharide treatment. Serum and cardiac tumor necrosis factor-alpha (TNF alpha) and interleukin-6 (IL-6) were detected by enzyme-linked immunosorbent assay methods at 1 h and 4.5 h after lipopolysaccharide treatment. Lipopolysaccharide treatment did not significantly affect the diabetic manifestations, including decreased body weight gain and increased glycated hemoglobin and serum triglyceride levels. However, diabetes significantly enhanced lipopolysaccharide-induced cardiac toxicity, which was demonstrated by significant increases in the levels of cardiac enzymes such as creatine phosphokinase and troponin T, abnormal morphological changes examined under light microscope with hematoxylin and eosin staining, and oxidative damage to proteins detected by 3-nitrotyrosine staining. Lipopolysaccharide treatment significantly increased serum and cardiac TNF-alpha and IL-6 concentrations. Diabetes did not alter the effect of lipopolysaccharide on serum and cardiac TNF-alpha elevation, but it significantly enhanced lipopolysaccharideinduced cardiac IL-6 production. These results suggest that diabetes significantly enhances endotoxin-induced cardiac toxicity, possibly through mechanisms that involve inflammatory/acute phase cytokines. PMID- 14734834 TI - Development of supporting materials for microbial immobilization and iron oxidation. AB - We developed the microbial immobilization particle with curdlan and activated carbon, which has great adsorption capacity. The characteristics of porosity and mechanical strength of these supporting particles are dependent on manufacturing method. The supporting particle showed the best performance when the ratio of curdlan and activated carbon was 30 to 6 g/L. Brumauer-Emmett-Teller (specific surface area) and swelling capacity of the carrier were 52.63 m2/g and 17 (w/w), respectively. The immobilization characteristics of iron-oxidizing bacteria on supporting particles were observed using a scanning electron microscope. The concentration of microorganism on the surface of supporting particle was increased with reaction time. As the number of iron oxidation batch cycles increased, the iron oxidation rate increased. PMID- 14734835 TI - Alkaline peroxide mechanical pulping of wheat straw with enzyme treatment. AB - Alkaline peroxide mechanical pulping (APMP) of wheat straw with enzyme treatment was studied. Instead of direct enzyme pretreatment on wheat straw, an alternative treatment method was used, in which coarse pulps from refiner defibrated wheat straw rather than wheat straw were pretreated with a crude enzyme containing mainly xylanase, then impregnated with alkaline H2O2 solution and further refined. The optimum conditions of enzyme treatment were xylanase dosage of 10-15 IU/g of oven-dried wheat straw, 90 min, 50-60 degrees C, pulp consistency of 5 10%, and initial pH of 5.0, and those for chemical impregnation were 6% NaOH, 70 80 degrees C, 60-90 min, and 4 to 5% H2O2. Enzyme treatment improved pulpability of wheat straw by the APMP process, and final pulp quality such as brightness, breaking length, and burst index of pulp. Pulp from the APMP process with enzyme treatment could be bleached to a brightness of 70.5% ISO by two-stage H2O2 bleaching sequence with only 4% H2O2, and breaking length of the bleach pulp reached 4470 m PMID- 14734836 TI - High-temperature wine making using the thermotolerant yeast strain Kluyveromyces marxianus IMB3. AB - Kluyveromyces marxianus IMB3 yeast cells were immobilized on delignified cellulosic material, apple, and quince separately. Both immobilized and free cells were used in high-temperature wine making, and their fermented grape must contained 3 to 4% alcohol. Semisweet wines were produced by the addition of potable alcohol to the fermented must. Preliminary sensory evaluation of the produced semisweet wines showed good flavor and aroma. The final product contained extremely low levels of higher and amyl alcohols while ethyl acetate was at levels usually present in wines. The ferment produced may be blended with other products to improve their quality. PMID- 14734837 TI - Production of beta-carotene from beet molasses by Blakeslea trispora in stirred tank and bubble column reactors: development of a mathematical modeling. AB - The effect of aeration rate and agitation speed on beta-carotene production from molasses by Blakeslea trispora in a stirred-tank fermentor and optimization of the production of the pigment in a bubble column reactor were investigated. In addition, a central composite design was employed to determine the maximum beta carotene concentration at optimum values for the process variables (aeration rate, sugar concentration, linoleic acid, kerosene). By image analysis of the morphology of the fungus, a quantitative characterization of the hyphae and zygospores formed was obtained. The hyphae were differentiated to intact hyphae, vacuolated hyphae, evacuated cells and degenerated hyphae. An increased proportion of zygospores was correlated to high beta-carotene production. In the stirred-tank fermentor, the highest concentration of the carotenoid pigment (92.0 mg/L) was obtained at an aeration rate of 1.5 vvm and agitation speed of 60 rpm. In the bubble column reactor, the aeration rate and concentration of sugars, linoleic acid, kerosene, and antioxidant significantly affected the production of beta-carotene. In all cases, the fit of the model was found to be good. Aeration rate, sugar concentration, linoleic acid, and kerosene had a strong positive linear effect on beta-carotene concentration. Moreover, the concentration of the pigment was significantly influenced by the negative quadratic effects of the given variables and by their positive or negative interactions. Maximum beta carotene concentration (360.2 mg/L) was obtained in culture grown in molasses solution containing 5% (w/v) sugar supplemented with linoleic acid (37.59 g/L), kerosene (39.11 g/L), and antioxidant (1.0 g/L). PMID- 14734838 TI - Lipase-catalyzed solvent-free transesterification of wood sterols. AB - Eighteen commercial lipase preparations, either immobilized or crude enzyme powders, were screened for the transesterification of wood sterols. The reactions were carried out in a solvent-free system, at the optimum temperature of the enzyme preparations as reported by the manufacturer and at the pressure of 2 mbar, with 5 or 10% in weight of the enzyme relative to the wood sterol content of the reacting mixture. Methyl esters of sunflower fatty acids were used as transesterifying agent. Of all the enzymes assayed, only Lipase TL from Pseudomonas stutzeri PL-836 (Meito Sangyo) exhibited any significant transesterifying capacity, 85 and 95% of conversion after 2 and 8 h of reaction, respectively, when 10% in weight of enzyme was used. PMID- 14734839 TI - Local and long-range order in ferroelastic lead phosphate at high pressure. AB - Pure lead phosphate, Pb(3)(PO(4))(2), undergoes a phase transition from C2/c to R(-)3m symmetry at a pressure of approximately 1.8 GPa and room temperature. Single-crystal X-ray diffraction measurements of the unit-cell parameters of a sample doped with 1.6% Ba(2+) for the Pb(2+) indicates that the doping reduces the transition pressure by approximately 0.1 GPa. The structural evolution of both samples through the phase transition has been determined by Rietveld refinement of neutron powder diffraction data collected to pressures of 6.3 and 3.3 GPa, respectively. There is no evidence for any significant change in the local structure at the phase transition at high pressures; the structure of the R(-)3m phase at pressures just above the phase transition includes disordered positions for several atoms. The observation of diffuse scattering from the R( )3m phase at high pressure by single-crystal X-ray diffraction suggests that the disorder is static and arises from the presence of several orientations of the ordered microdomains of the monoclinic local structure. The macroscopic transition from monoclinic to trigonal symmetry therefore appears to correspond to the pressure at which the coherency strains between the locally monoclinic microdomains are sufficient to create a dimensionally trigonal lattice within which local displacements of atoms are still significant. A further pressure increase then decreases the magnitude of these displacements until at 3.5 GPa or higher they are not detectable by our current experimental probes, and the structure appears to have true local and global trigonal symmetry. PMID- 14734840 TI - Jahn-Teller distortions, cation ordering and octahedral tilting in perovskites. AB - In transition metal oxides, preferential occupation of specific d orbitals on the transition metal ion can lead to the development of a long-range ordered pattern of occupied orbitals. This phenomenon, referred to as orbital ordering, is usually observed indirectly from the cooperative Jahn-Teller distortions (CJTDs) that result as a consequence of the orbital ordering. This paper examines the interplay between orbital ordering, octahedral tilting and cation ordering in perovskites. Both ternary AMX(3) perovskites containing an active Jahn-Teller (J T) ion on the octahedral site and quaternary A(2)MM'X(6) perovskites containing a J-T ion on one-half of the octahedral sites have been examined. In AMX(3) perovskites, the tendency is for the occupied 3d(3x2-r2) and 3d(3z2-r2) orbitals to order in the ac plane, as exemplified by the crystal structures of LaMnO(3) and KCuF(3). This arrangement maintains a favorable coordination environment for the anion sites. In AMX(3) perovskites, octahedral tilting tends to enhance the magnitude of the J-T distortions. In A(2)MM'X(6) perovskites, the tendency is for the occupied 3d(3z2-r2) orbitals to align parallel to the c axis. This pattern maintains a favorable coordination environment about the symmetric M'-cation site. The orbital ordering found in rock-salt ordered A(2)MM'X(6) perovskites is compatible with octahedral rotations about the c axis (Glazer tilt system a(0)a(0)c(-)) but appears to be incompatible with GdFeO(3)-type octahedral tilting (tilt system a(-)b(+)a(-)). PMID- 14734841 TI - Superspace description of NaCa4Nb5O17 (a perovskite-related compound of the type AnBnO3n+2) as a modulated layered structure. AB - The recently determined structure of NaCa(4)Nb(5)O(17) is reanalyzed within the superspace framework. The material follows the general features of the superspace model proposed for the perovskite-related compounds of the general formula A(n)B(n)O(3n+2) [Elcoro et al. (2001). Acta Cryst. B57, 471-484]. It can be described as a commensurate modulated structure with discontinuous atomic domains given by occupational crenel functions. These atomic domains automatically introduce the layered configuration of the actual structure in real space. However, the displacive modulations follow a quite different pattern from that observed in the Sr(n)(Nb,Ti)(n)O(3n+2) series, discussed in the above-mentioned reference. The superspace group relevant in this new case has been identified through a systematic search of all the possible groups and a comparison of the resulting superspace embeddings of the experimental three-dimensional structure. Being a commensurate structure, the fundamental ambiguity of the superspace description was broken by choosing the highest possible symmetry that yields smooth displacive atomic modulations and hence minimizes the number of parameters. The efficiency of the proposed superspace model is demonstrated with a new refinement of the structure. Assuming the model's general relevance, symmetry properties for the whole [Ca,Na](n)Nb(n)O(3n+2) series are predicted. PMID- 14734842 TI - Structures and phase transitions of trigonal ZrMo2O8 and HfMo2O8. AB - This paper describes the structures, thermal-expansion properties and phase transitions of the trigonal forms of ZrMo(2)O(8) and HfMo(2)O(8). Both phases adopt a P(-)3m structure at room temperature and show positive thermal expansion. Both phases also undergo a displacive phase transition at high temperature (ZrMo(2)O(8) at 487 K and HfMo(2)O(8) at 463 K) to a higher-symmetry structure that has lower thermal expansion. The structure of the high-temperature alpha' AMo(2)O(8) form (A = Zr and Hf) has been refined from powder diffraction data in space group P(-)3m1. PMID- 14734843 TI - An X-ray powder diffraction study of the spin-crossover transition and structure of bis(2,6-dipyrazol-1-ylpyrazine)iron(II) perchlorate. AB - The crystal structure of the iron(II) spin-crossover compound [Fe(C(10)H(8)N(6))(2)](ClO(4))(2) in the high-spin state has been solved from powder X-ray diffraction data using the DASH program and refined using Rietveld refinement. The thermal spin transition has been monitored by following the change in unit-cell parameters with temperature. The title compound has been found to undergo a crystallographic phase change, involving a doubling of the crystallographic a axis, on undergoing the spin transition. PMID- 14734845 TI - Adducts of 1,4,8,11-tetraazacyclotetradecane with carboxylic acids: hydrogen bonded supramolecular structures in two or three dimensions. AB - Four solvated salt-type adducts derived from cyclam (1,4,8,11 tetraazacyclotetradecane) and carboxylic acids have been structurally characterized. In the salt derived from adamantane-1-carboxylic acid, 4,11-diaza 1,8-diazoniacyclotetradecane bis(adamantane-1-carboxylate) tetrahydrate, (1) (monoclinic, P2(1)/c, Z' = 0.5), where the cation lies across a centre of inversion, the anions and the water molecules form chains of edge-fused R(4)(2)(8) and R(6)(6)(16) rings, which are linked into sheets by the cations. In the 4-aminobenzoate salt, 4,11-diaza-1,8-diazoniacyclotetradecane bis(4 aminobenzoate) monohydrate, (2) (monoclinic, C2/c, Z' = 0.5), where the cation lies across a centre of inversion and the water molecule lies across a twofold rotation axis, the cations and anions generate a three-dimensional framework, readily analysed in terms of two distinct two-dimensional substructures, viz. (10;1) sheets of R(8)(6)(46) rings, and pairwise interwoven (100) sheets, reinforced by water molecules. The 3-hydroxybenzoate salt, 4,11-diaza-1,8 diazoniacyclotetradecane bis(3-hydroxybenzoate) dihydrate, (3) (monoclinic, Pc, Z' = 1), contains a three-dimensional framework constructed from anions and water molecules only, which encapsulates large voids and within which the cations are linked to the anion-water framework via N-H.O hydrogen bonds. There are two independent cations in 4,11-diaza-1,8-diazoniacyclotetradecane 5 hydroxyisophthalate(2-) methanol solvate, (4) (monoclinic, P2(1)/c, Z' = 1), both lying across centres of inversion but with entirely different configurations. The anions alone form simple chains, and these chains are linked by the two types of cation into a three-dimensional framework from which the methanol molecules are pendent. Comparisons are made with carboxylate complexes of the [Ni(cyclam)](2+) cation and with carboxylate salts derived from meso-5,5,7,12,12,14-hexamethyl 1,4,8,11-tetraazacyclotetradecane. PMID- 14734844 TI - Hydrated metal(II) complexes of N-(6-amino-3,4-dihydro-3-methyl-5-nitroso-4 oxopyrimidin-2-yl) derivatives of glycine, glycylglycine, threonine, serine, valine and methionine: a monomeric complex and coordination polymers in one, two and three dimensions linked by hydrogen bonding. AB - Nine hydrated complexes of Group 2 (alkaline earth) cations with organic ligands which are N-substituted amino acids containing the 6-amino-3,4-dihydro-3-methyl-5 nitroso-4-oxopyrimidin-2-yl group have been structurally characterized. The octahydrated calcium glycinate complex, where the six-coordinate Ca cation lies on an inversion centre in the space group P(-)1, forms a finite (zero dimensional) complex. The hexahydrated barium glycinate complex contains eight coordinate Ba and it is isostructural with the known Sr analogue, and its one dimensional coordination polymer takes the form of a simple chain. The octahydrated calcium and strontium threonine complexes are isostructural, with eight-coordinate cations lying on twofold rotation axes in the space group C2: the one-dimensional coordination polymers take the form of a chain of spiro-fused rings and a similar chain of spiro-fused rings is found in the heptahydrated barium serine complex, although here the ten-coordinate cation lies in a general position. In the tetrahydrated strontium and barium glycylglycinate complexes, the eight-coordinate cations lie on twofold rotation axes in the space group C2/c, but in the Sr complex the coordination polymer is a chain of spiro-fused rings, while in the Ba complex the coordination polymer forms deeply puckered sheets. There are two types of Ca site in the hexahydrated calcium valine complex: one is eight coordinate and gives rise to a two-dimensional coordination polymer, while the other is seven coordinate forming a finite, zero-dimensional coordination complex. In the heptahydrated barium methionine complex, the coordination polymer is three dimensional. In all of the complexes, the coordination aggregates are further linked by an extensive series of hydrogen bonds. PMID- 14734846 TI - Symmetrically 4,6-disubstituted 2-aminopyrimidines and 2-amino-5 nitrosopyrimidines: interplay of molecular, molecular-electronic and supramolecular structures. AB - The structures of six symmetrically 4,6-disubstituted 2-aminopyrimidines, four of them containing a 5-nitroso substituent, have been determined. The nitroso compounds, in particular, exhibit polarized molecular-electronic structures leading to extensive charge-assisted hydrogen bonding. The intermolecular interactions observed include hard hydrogen bonds of N-H...N and N-H...O types together with O-H...O and O-H...N types in 2-amino-4,6-bis(2-hydroxyethylamino)-5 nitrosopyrimidine; soft hydrogen bonds of the C-H...O type in both 2-amino-4,6 bis(morpholino)-5-nitrosopyrimidine (3) and 2-amino-4,6-bis(benzylamino)-5 nitrosopyrimidine (4), and of the C-H...pi(arene) type in both 2-amino-4,6 bis(piperidino)pyrimidine (1) and 2-amino-5-nitroso-4,6-bis(3 pyridylmethoxy)pyrimidine (5); and aromatic pi...pi stacking interactions in 2 amino-5-nitroso-4,6-bis(3-pyridylmethoxy)pyrimidine. The supramolecular structures formed by the hard hydrogen bonds are finite, zero-dimensional in (1), one-dimensional in 2-amino-4,6-bis(3-pyridylmethoxy)pyrimidine (2), two dimensional in both (3) and (4), and three-dimensional in both (5) and 2-amino 4,6-bis(2-hydroxyethylamino)-5-nitrosopyrimidine. PMID- 14734847 TI - Supramolecular assembling using synthons with NH-CO(S)-CS-NH and NH-CO-CO-NH functionalities: crystal structures of (S,S)-N,N'-monothiooxalyldileucine methyl ester and its dithio analogue. AB - To compare the structural properties of oxalamide and thiooxalamide groups in the formation of hydrogen bonds suitable for supramolecular assemblies a series of retropeptides was studied. Some of them, having oxalamide bridges, are gelators of organic solvents and water. However, retropeptides with oxygen replaced by the sp(2) sulfur have not exhibited such properties. The crystal structures of the two title compounds are homostructural, i.e. they have similar packing arrangements. The monothio compound crystallizes in the orthorhombic space group P2(1)2(1)2(1) with two molecules in the asymmetric unit arranged in a hydrogen bond network with an approximate 4(1) axis along the crystallographic b axis. However, the dithio and dioxo analogues crystallize in the tetragonal space group P4(1) with similar packing patterns and hydrogen-bonding systems arranged in agreement with a crystallographic 4(1) axis. Thus, these two analogues are isostructural having closely related hydrogen-bonding patterns in spite of the different size and polarity of oxygen and sulfur which serve as the proton acceptors. PMID- 14734848 TI - Different packing in three polymorphs of 2,4,6-trimethoxy-1,3,5-triazine. AB - 2,4,6-trimethoxy-1,3,5-triazine was found to exhibit three different polymorphs. The alpha-polymorph undergoes reversible phase transformation to the beta polymorph at 340 K with an enthalpy of 3.9 kJ mol(-1). The heat of fusion of the beta-polymorph at 404 K is 18.1 kJ mol(-1). The low-temperature phase (alpha polymorph) crystallizes in the orthorhombic space group Pnma. The high temperature phase (beta-polymorph) can be obtained from the melt and crystallizes in the monoclinic space group P2(1). The gamma-polymorph is obtained by crystallization from a 1:1 mixture of methanol and methylenechloride from hydrolyzed 2,4-dimethoxy-6-oxybenzophenone-1,3,5-triazine. The gamma-polymorph melts at 409 K with an enthalpy of 11.4 kJ mol(-1). The gamma-polymorph crystallizes in the trigonal space group R3c. The molecules occupy a crystallographic threefold axis. Molecules of 2,4,6-trimethoxy-1,3,5-triazine are planar in all three polymorphs. The major difference between the three polymorphs is the mode of packing of the molecules in the crystal. PMID- 14734849 TI - Geometry at the aliphatic tertiary carbon atom: computational and experimental test of the Walsh rule. AB - The geometrical parameters of molecules of 2-substituted 2-methylpropanes and 1 substituted bicyclo[2.2.2]octanes were calculated at the B3LYP/6-311+G(d,p) level. They agreed reasonably well with the mean crystallographic values retrieved from the Cambridge Structural Database for a set of diverse non-cyclic structures with a tertiary C atom. The angle deformations at this C atom produced by the immediately bonded substituent are also closely related to those observed previously in benzene mono derivatives (either as calculated or as derived from crystallographic data). The calculated geometrical parameters were used to test the classical Walsh rule: It is evidently true that an electron-attracting substituent increases the proportion of C-atom p-electrons in the bond to the substituent and leaves more s-electrons to the remaining bonds; as a consequence the C-C-C angles at a tertiary carbon are widened and the C-C bonds shortened. However, this rule describes only part of the reality since the bond angles and lengths are controlled by other factors as well, for instance by steric crowding. Another imperfection of the Walsh rule is that the sequence of substituents does not correspond to their electronegativities, as measured by any known scale; more probably it is connected with the inductive effect, but then only very roughly. PMID- 14734850 TI - Quininium (R)-mandelate, a structure with large Z' described as an incommensurately modulated structure in (3+1)-dimensional superspace. AB - Quininium (R)-mandelate, C(20)H(25)N(2)O(2)(+).C(8)H(7)O(3)(-), is an organic salt with an incommensurately modulated structure. The superspace approach is required for a precise description of the structure at room temperature. In addition to the main reflections, the diffraction pattern also exhibits higher order satellite reflections. The large number of first- and second-order satellite reflections with relatively strong intensities indicates a significant modulation. No average structure solution could be obtained on the basis of the main reflections only and hence the structure solution was performed in two suitable superstructure approximations, including satellite intensities. Positional modulation functions and modulation functions for the anisotropic atomic displacement parameters were introduced. The modulation originates from a competition between intramolecular and intermolecular forces, which is reflected, in the superspace formalism, in the modification of hydrogen bonds along the internal space variable t. PMID- 14734852 TI - Evaluation of three different forward primers by terminal restriction fragment length polymorphism analysis for determination of fecal bifidobacterium spp. in healthy subjects. AB - The 27F forward primer is frequently used in 16S rRNA gene libraries and T-RFLP analysis. However, Bifidobacterium spp. were barely detected with this primer in human fecal samples. In this study, fecal microbiota were analyzed using the T RFLP method with three different forward primers (27F, 35F, and 529F) in conjunction with one reverse primer (1492R). T-RFLP analysis of fecal microbiota using 35F and 529F detected higher proportions of the terminal restriction fragment (T-RF) corresponding to Bifidobacterium spp. than that using 27F. 27F is in imperfect agreement while 35F and 529F are in good concordance with the 16S rRNA gene sequences of Bifidobacterium spp., and the latter primers allowed for the detection of T-RFs of Bifidobacterium spp. in fecal samples from five healthy subjects. The T-RFs presumed to be Bifidobacterium spp. were cloned and sequenced, and found to match the 16S rRNA gene sequences of Bifidobacterium spp. Among the five fecal samples, two samples with low frequencies of T-RFs of Bifidobacterium spp. were detected using these forward primers. This probably reflects a low prevalence of Bifidobacterium spp. in these two samples. Our study emphasizes the importance of selecting a suitable forward primer for detection of Bifidobacterium spp. PMID- 14734853 TI - Effects of tacrolimus (FK506) on encephalomyocarditic virus-induced diabetes in mice. AB - The effects of tacrolimus on insulin-dependent diabetes mellitus (IDDM) induced by the D-variant of encephalomyocarditis virus (D-EMCV) have been investigated. Male BALB/c mice were treated with tacrolimus before viral inoculation, and then were inoculated with 10 plaque forming units (PFU) of DEMCV. The mice continued to be treated with tacrolimus until the animals were sacrificed. D-EMCV-infected mice, which were treated with saline as controls, showed abnormal glucose tolerance test (GTT) values, whereas all infected mice with tacrolimus pretreatment were normal on 7 days-post inoculation (DPI). Histological observations revealed that non-treated tacrolimus D-EMCV-infected mice and which developed diabetes showed severe insulitis in their islets of Langerhans. On the other hand, D-EMCV-infected mice treated with tacrolimus were normal. In D-EMCV infected mice, viruses in the pancreata were detected at the same level regardless of treatment with tacrolimus or saline. Expressions of TNF-alpha and IFN-gamma mRNA in spleens of tacrolimus-treated D-EMCV-infected mice were lower than that of non-treated tacrolimus DEMCV-infected mice on 7 DPI. The results suggest that tacrolimus suppresses expressions of TNF-alpha and IFN-gamma mRNAs to prevent the onset of D-EMCV-induced IDDM. PMID- 14734854 TI - Differential susceptibility of cells expressing allogeneic MHC or viral antigen to killing by antigen-specific CTL. AB - CD8(+) cytotoxic T lymphocytes (CTLs) generated by immunization with allogeneic cells or viral infection are able to lyse allogeneic or virally infected in vitro cells (e.g., lymphoma and mastocytoma). In contrast, it is reported that CD8(+) T cells are not essential for allograft rejection (e.g., heart and skin), and that clearance of influenza or the Sendai virus from virus-infected respiratory epithelium is normal or only slightly delayed after a primary viral challenge of CD8-knockout mice. To address this controversy, we generated H-2(d)-specific CD8(+) CTLs by a mixed lymphocyte culture and examined the susceptibility of a panel of H-2(d) cells to CTL lysis. KLN205 squamous cell carcinoma, Meth A fibrosarcoma, and BALB/c skin components were found to be resistant to CTL mediated lysis. This resistance did not appear to be related to a reduced expression of MHC class I molecules, and all these cells could block the recognition of H-2(d) targets by CTLs in cold target inhibition assays. We extended our observation by persistently infecting the same panel of cell lines with defective-interfering Sendai virus particles. The Meth A and KLN205 lines infected with a variant Sendai virus were resistant to lysis by Sendai virus specific CTLs. The Sendai virus-infected Meth A and KLN205 lines were able to block the lysis of Sendai virus-infected targets by CTLs in cold target inhibition assays. Taken together, these results suggest that not all in vivo tissues may be sensitive to CTL lysis. PMID- 14734856 TI - Detection of antigen-specific T cells in experimental immune-mediated blepharoconjunctivitis in DO11.10 T cell receptor transgenic mice. AB - Antigen (Ag)-specific T cells are thought to play a key role in pathogenesis of chronic allergic conjunctivitis (AC) such as atopic keratoconjunctivitis (AKC) and vernal keratoconjunctivitis (VKC). In order to investigate the trafficking of Ag-specific T cells in experimental immune-mediated blepharoconjunctivitis (EC), we established a novel AC model in DO11.10 T cell receptor (TcR) transgenic (Tg) mice. DO11.10 TcR-Tg mice were challenged with eye drops of whole OVA protein, OVA peptide 1-15, 321-335, or 323-339. Their eyes were histologically examined. Conventional proliferation assay was performed against each Ag. Phenotypes of infiltrating cells and kinetics of Ag-specific T cells were investigated by immunohistochemistry. Adoptive transfer of CD4(+) Ag-specific T cells from DO11.10 TcR-Tg to WT mice was performed. The distribution of KJ1-26(+) cells was investigated in recipient mice. The challenge of OVA peptide 323-339 induced infiltration of inflammatory cells in conjunctivae in a dose dependent manner, accompanied by the proliferative responses of splenocytes. Immunohistochemical analysis revealed Agspecific/ non-Ag-specific T cells, macrophages, and eosinophils in conjunctivae. Infiltration of Ag-specific T cells increased 24 hr later. Transfer of CD4(+) cells from DO11.10 TcR-Tg to WT mice induced EC depending on the number of transferred cells. Ag-specific T cells were detected in the conjunctivae and spleens of recipient mice, though its numbers were significantly smaller compared to DO11.10 TcR-Tg mice. The challenge of OVA peptide 323-339 induced EC in DO11.10 TcR-Tg mice without prior sensitization. The response was mediated by CD4(+) Ag-specific T cells. The trafficking of Ag specific T cells in EC was clearly visualized. PMID- 14734855 TI - Homologous and heterologous antibody responses to lipopolysaccharide after enterohemorrhagic Escherichia coli infection. AB - To evaluate antibody responses against lipopolysaccharide (LPS: O157, O26, and O111) in enterohemorrhagic Escherichia coli(EHEC) infection, sera of 24 schoolchildren associated with the Morioka outbreak in 1997 and of 74 sporadic patients suspected of having EHEC infection were examined. Using a positive standard serum, quantitative evaluation of LPS antibodies by an enzyme-linked immunosorbent assay (ELISA) was established. High levels of specific IgM and IgA antibodies against homologous E. coli LPS were present in the acute period and are characteristic of EHEC. This could be used for the serological diagnosis of EHEC infection, except for early infants and the elderly. In addition to the specific homologous response, multiple antibody responses against different serotypes other than those isolated were demonstrated in many cases by qualitative analysis using Western blotting. PMID- 14734857 TI - A multi-prefectural outbreak of Shigella sonnei infections associated with eating oysters in Japan. AB - Among roughly one thousand incidents of shigellosis annually in Japan, approximately 70% of the cases are estimated to be associated with overseas travel. However, at the end of 2001, reports of domestically acquired Shigella sonnei infections suddenly increased. We report here the first multi-prefectural outbreak of Shigella sonnei infections linked to the consumption of imported oysters in Japan at the end of 2001. Isolates of S. sonnei from patients epidemiologically linked to eating contaminated oysters and from the imported oysters themselves showed an indistinguishable pulsed-field gel electrophoresis pattern and drug resistance pattern. PMID- 14734858 TI - Contribution of a single hydrogen bond between betaHis81 of MHC class II I-E(k) and the bound peptide to the pH-dependent thermal stability. AB - To determine the energetic contribution of the hydrogen bond between betaHis81 of the major histocompatibility complex class II (MHC II) molecule, I-E(k), and the bound hemoglobin peptide (Hb), we analyzed the thermal stability of the hydrogen bond-disrupted mutant, I-E(k)-Hb betaH81Y, in which the betaHis81 residue was replaced with Tyr, by differential scanning calorimetry. The thermal stability of the I-E(k)-Hb betaH81Y mutant was lower than that of the I-E(k)-Hb wild-type, mainly due to the decreased enthalpy change. The difference in the denaturation temperature of the I-E(k)-Hb betaH81Y mutant as compared with that of the I-E(k) Hb wild-type at pH 5.5 was only slightly smaller than that at pH 7.4, in agreement with the increased stability at an acidic pH, a unique characteristic of MHC II. Thus, the hydrogen bond contributed by betaHis81 is critical for peptide binding, and is independent of pH, which can alter the hydrophilicity of the His residue. PMID- 14734859 TI - CD72 stimulation modulates anti-IgM induced apoptotic signaling through the pathway of NF-kappaB, c-Myc and p27(Kip1). AB - Engagement of mIgM induces G1 arrest and apoptosis in immature B cells. The biochemical mechanism(s) regulating the cell death process are poorly understood. Cross-linking of CD72 (a B cell co-receptor) with anti-CD72 antibody was shown to protect B cells from apoptosis. We investigated the molecular mechanism involved in apoptosis preventing signaling mediated by CD72 ligation using a derivative (WEHIdelta) of the WEHI231 cell line which is representative of immature B cells. Apoptotic WEHIdelta cells following cross-linking of mIgM demonstrate a dramatic loss of c-Myc protein after transient up-regulation. In contrast, pre-ligation of CD72 was able to sustain c-Myc expression after transient up-regulation. Cross linking of mIgM of WEHIdelta cells causes accumulation of the Cdk inhibitor, p27(Kip1). CD72 pre-ligation was shown to inhibit the accumulation of p27(Kip1) protein. Moreover, NF-kappaB activity was not suppressed in WEHIdelta cells after mIgM cross-linking when the cells were pre-treated with anti-CD72 antibody. These results strongly suggest that the apoptosis preventing signal evoked by CD72 ligation is delivered through the pathway of NF-kappaB, c-Myc, p27(Kip1) and cyclin. PMID- 14734860 TI - Mechanisms of action of corilagin and tellimagrandin I that remarkably potentiate the activity of beta-lactams against methicillin-resistant Staphylococcus aureus. AB - Corilagin and tellimagrandin I are polyphenols isolated from the extract of Arctostaphylos uvaursi and Rosa canina L. (rose red), respectively. We have reported that corilagin and tellimagrandin I remarkably reduced the minimum inhibitory concentration (MIC) of beta-lactams in methicillin-resistant Staphylococcus aureus(MRSA). In this study, we investigated the effect of corilagin and tellimagrandin I on the penicillin binding protein 2 '(2a) (PBP2 '(PBP2a)) which mainly confers the resistance to beta-lactam antibiotics in MRSA. These compounds when added to the culture medium were found to decrease production of the PBP2 '(PBP2a) slightly. Using BOCILLIN FL, a fluorescent labeled benzyl penicillin, we found that PBP2 '(PBP2a) in MRSA cells that were grown in medium containing corilagin or tellimagrandin I almost completely lost the ability to bind BOCILLIN FL. The binding activity of PBP2 and PBP3 were also reduced to some extent by these compounds. These results indicate that inactivation of PBPs, especially of PBP2 '(PBP2a), by corilagin or tellimagrandin I is the major reason for the remarkable reduction in the resistance level of beta-lactams in MRSA. Corilagin or tellimagrandin I suppressed the activity of beta-lactamase to some extent. PMID- 14734861 TI - Reflections on shunt infection. AB - The placement and revision of ventriculoperitoneal (VP) shunts remains a mainstay in the surgical treatment of hydrocephalus. While the North American infection rate averages nearly 8-10%, published infection rates for VP shunt infection below 1% have been reported. We retrospectively reviewed shunt operations by a single surgeon over 62 months to analyze the infection rate. In 62 months, we performed 526 shunt placements or revisions in patients up to 18 years of age. There were 7 shunt infections (1.33%). In 5 cases, the organism was Staphylococcus epidermidis, and a single shunt each was infected with Haemophilus influenzae and Staphylococcus aureus. Each infection was treated with external ventriculostomy drainage and intravenous antibiotics. The new shunt was placed at a new incision site after at least 5 days of sterile spinal fluid cultures. The mean follow-up among these patients after shunt insertion was 25 months. VP shunting remains the most common operation for hydrocephalus. Infections are linked with seizures, higher future risks of shunt infection and malfunction, and reduced IQ and school performance. Our infection rate during 62 months was limited to 1.33%. Uniform surgical technique, limited hardware and skin edge manipulation and double gloving may be important factors in limiting shunt infections. PMID- 14734862 TI - Treatment of spinal involvement in neuroblastoma patients. AB - INTRODUCTION: Considerable controversy exists regarding the appropriate management of spinal involvement in neuroblastoma (NB) patients. We review a large group of such patients and offer treatment recommendations. METHODS: Forty six patients with epidural and/or neural foraminal involvement treated between 1987 and 1998 were staged according to the International NB Staging System (INSS) and classified as high-risk (INSS stage 4; n = 31) or low-risk (INSS stage <4; n = 15). Of 13 high- risk patients with normal neurologic examinations and no radiographic high-grade spinal cord compression (HGSCC), 12 were treated initially with chemotherapy, and only 1 demonstrated neurologic deterioration. HGSCC was present in 18 patients with high-risk NB; 7 of 10 (70%) treated initially with chemotherapy and 6 of 6 (100%) managed initially with operation improved or remained stable. All 9 low-risk patients with normal neurologic examinations and no HGSCC remained neurologically intact following operations (n = 7) or chemotherapy (n = 2). All 4 low-risk patients with HGSCC treated with operations improved or remained stable, and 0 of 2 (0%) low-risk patients treated initially with chemotherapy remained stable. Spinal deformities occurred in 2 of 16 patients (12.5%) treated nonoperatively and in 9 of 30 (30.0%) who underwent operations. CONCLUSIONS: High-risk NB patients with spinal involvement but normal neurologic examinations should be offered chemotherapy. High-risk patients with HGSCC may respond to chemotherapy, but a small percentage will require operations for progressive neurologic deficits. Chemotherapy may be avoided in low-risk patients who are offered potentially curative operations. Patients treated with operations for epidural disease are at high risk of subsequently developing spinal deformity. PMID- 14734863 TI - Molecular biology of medulloblastoma therapy. AB - Molecular biological studies have created a new understanding of medulloblastomas, revealing key cell signaling pathways that promote tumor growth. These studies have identified molecular markers that can serve as prognostic indicators and which will provide therapeutic targets in the very near future. In contrast to conventional histological techniques, molecular biological methods are designed to detect highly unstable molecules degraded by conventional tissue collection and fixation protocols. Consequently, a new approach to tissue collection is required for molecular analysis as we enter the next era of brain tumor therapy. PMID- 14734864 TI - Lumbar split cord malformation and Klippel-Feil syndrome. AB - The authors report a patient with type II split cord malformation (SCM) and Klippel-Feil syndrome (KFS). Considering our review of the medical literature and present case report, we believe lumbar SCM and KFS in the same patient not to be coincidental findings but rather embryologically related entities. We propose that KFS in at least one of its phenotypes represents a forme fruste of occult spinal dysraphism. PMID- 14734865 TI - Tacrolimus and cyclosporine A are of no benefit to young rats with kaolin-induced hydrocephalus. AB - Hydrocephalus causes damage to periventricular axons. Tacrolimus, cyclosporine A (CsA) and calpain inhibitors have been shown to protect axons in rat models of acute traumatic brain injury. We hypothesized that these agents would ameliorate the axon damage and behavioral effects in experimental hydrocephalus. Hydrocephalus was induced in 3-week-old rats by injection of kaolin into the cisterna magna. Tests of cognitive and motor function were performed on a weekly basis. In a blinded and randomized manner, tacrolimus (FK506; 3.6 mg/kg body weight) or CsA (10 mg/kg) was administered once daily by subcutaneous injection for 2 weeks, beginning 2 weeks after induction of hydrocephalus. In a separate experiment, calpain inhibitor I (10 mg/kg/day) was administered by continuous subcutaneous infusion. The brains were subjected to histopathological and biochemical analyses after 2 weeks of treatment. There was no statistically significant protection in regard to behavior, brain structure or brain composition in any of the experiments. However, there was biochemical and histological evidence of renal injury following chronic tacrolimus and CsA administration. Calcineurin inhibition does not offer significant protection in this rat model of hydrocephalus. PMID- 14734866 TI - Surgical outcome following resection of contrast-enhanced pediatric brainstem gliomas. AB - BACKGROUND: The role of surgery in the management of gadolinium-enhancing pediatric brainstem lesions on magnetic resonance imaging (MRI) has been a matter of open debate. This clinical series correlates radiological and pathological findings to assess the role of contrast enhancement as an indication for surgery with respect to clinical outcome. METHODS: We retrospectively reviewed the charts of all pediatric patients admitted to the Johns Hopkins Hospital with a diagnosis of a brainstem tumor between January 1985 and December 2000. RESULTS: There were a total of 89 patients who met the inclusion criteria. Fifty-seven patients (64.0%) underwent surgical resection while 32 (36%) were treated with radiation and/or chemotherapy. Of the surgical candidates, 57 (100%) had an accompanying MRI scan significant for an enhancing lesion in the midbrain, pons or the medulla. The pathology was consistent with juvenile pilocytic astrocytoma in 30 patients (52.6%) and glioblastoma multiforme in 12 patients (21.1%). The remaining cases consisted of 10 patients (17.5%) with fibrillary astrocytomas, 3 (5.3%) with gangliogliomas, 1 (1.8%) with an oligodendroglioma and 1 (1.8%) with a primitive neuroectodermal tumor. A total of 29 patients had a total surgical resection, 8 a near total resection (>90%), 15 a subtotal resection (50-90%) and 5 a partial resection (<50%). The progression-free survival of all patients was 71.9% at 3 years and 45.6% at 5 years. CONCLUSIONS: This case series illustrates that contrast-enhanced MRI has positive prognostic value in the management of pediatric brainstem gliomas. In our study, the majority of enhancing tumors were low grade and amenable to surgical intervention. Consequently, we recommend surgical resection and pathological diagnosis of all enhancing brainstem tumors with adjuvant therapy reserved for recurrent or unresectable cases. PMID- 14734867 TI - Cystoventricular shunting of intracranial arachnoid cysts. AB - Ten patients with intracranial arachnoid cysts were treated with direct shunting of the cyst to a lateral ventricle. The strategic goal of cystoventricular shunting is to establish physiologically normal intracranial pressure relationships, rather than cyst obliteration. Cystoventricular shunts were successful in treating single and multiple intracranial cysts in supratentorial and infratentorial locations and in patients with normal and enlarged lateral ventricles. Cystoventricular shunting is conceptually simple as well as effective and reliable. PMID- 14734868 TI - Distances from the atlantal segment of the vertebral artery to the midline in children. AB - There are scant data in the literature regarding distances from the midline to the suboccipital segment of the vertebral artery, and to our knowledge, none addressing this measurement in the pediatric population. This study seeks to establish age-related distances (Y) from the laterally placed vertebral artery to the midline at the level of the posterior arch of the atlas in children. Measurements from the midline to the most medial portion of the vertebral artery at the level of the posterior arch of the atlas were made in 50 children who underwent CT imaging of the cervical spine. For all left sides, Y = 12-23 mm (mean 17 mm). For all right sides, Y = 10-25 mm (mean 17.5 mm). For girls, Y = 12 16 mm (mean 12.7 mm) for left sides and Y = 12-25 mm (mean 18.5 mm) for right sides. For boys, Y = 12-20 mm (mean 16 mm) for left sides and Y = 10-21 mm (mean 16.7 mm) for right sides. We have found that the most medial portion of the vertebral artery from the midpoint of the posterior arch of the atlas for all ages was approximately 17 and 17.5 mm for left and right sides, respectively. This distance was generally less for left sides, except in the 16-19 years age group. However, as a group, this distance did not vary more than 2.8 mm between left and right sides. These data should assist the neurosurgeon who operates in the suboccipital region in children. PMID- 14734869 TI - Spontaneous intracerebral hemorrhage and multiple infarction in Williams-Beuren syndrome. AB - A 7-year-old boy diagnosed with Williams-Beuren syndrome was admitted for spontaneous right hemispheric intracerebral hemorrhage. Cerebral angiography did not reveal any source of bleeding. After a short period of clinical improvement under conservative treatment, the boy deteriorated rapidly. CT showed the beginning of a complete infarction of both hemispheres. Operative evacuation of the bleeding and bilateral osteoclastic decompression had no perceptible influence on the clinical course. To the present day, the boy has remained in a vegetative state. Reports in the literature suggest that Williams syndrome with cerebral infarction is associated with a markedly poorer prognosis when there is additional intracerebral bleeding. PMID- 14734870 TI - Traumatic retroclival epidural hematoma in a child. PMID- 14734871 TI - Interaction between resident luminal bacteria and the host: can a healthy relationship turn sour? PMID- 14734872 TI - Acid and iron-disturbances related to Helicobacter pylori infection. PMID- 14734873 TI - Gluten-free diet, chromosomal abnormalities, and cancer risk in coeliac disease. PMID- 14734874 TI - Tackling moderate malnutrition: the next frontier. PMID- 14734875 TI - Gastric histopathology, iron status and iron deficiency anemia in children with Helicobacter pylori infection. AB - OBJECTIVES: Helicobacter pylori has been established as a major cause of gastritis and peptic ulcer disease in adults and children. H. pylori infection may also have a role in the development of some extra-gastrointestinal diseases, including iron deficiency anemia. The aim of this study is to investigate H. pylori-related changes in gastric physiology and histology and the relationship of these changes to iron deficiency anemia in children. METHODS: Fifty-two patients with gastrointestinal complaints were studied. Hematologic parameters, 3 day vitamin C and iron consumption, serum gastrin levels, and gastric juice ascorbic acid levels were compared in patients with and without H. pylori infection. Dietary intake of vitamin C and iron, serum gastrin, gastric juice ascorbic acid content, and gastric histology were compared in patients with H. pylori infection and anemia and in patients with H pylori infection and no anemia. The CagA status of the H. pylori organisms was evaluated. RESULTS: Twenty eight of 52 patients had H. pylori. Thirty-one patients had iron deficiency anemia. H. pylori infection was associated with low serum iron levels. H. pylori gastritis was associated with a decrease in the gastric juice ascorbic acid level. Infection with CagA-positive strains was associated with a greater decrease in gastric juice ascorbic acid than infection with CagA-negative strains. However, the gastric juice ascorbic acid levels of patients with H. pylori and anemia were not different from those of non-anemic patients with H. pylori. Among patients with H. pylori infection, pangastritis was twice as common in those with anemia than in those without anemia. CONCLUSIONS: H. pylori infection was associated with a decrease in gastric juice ascorbic acid concentration, and this effect was more pronounced in patients with the CagA positive strain. Pangastritis was more common in patients whose H. pylori.infection was accompanied by anemia. PMID- 14734876 TI - Supplementary feeding of underweight, stunted Malawian children with a ready-to use food. AB - OBJECTIVE: Maize and soy flour mixes are often used in the treatment of moderate malnutrition in Malawi. Their efficacy has not been formally evaluated. A recently developed ready-to-use food (RTUF) effectively promotes growth among severely malnourished children. The authors compared the effect of maize and soy flour with that of RTUF in the home treatment of moderately malnourished children. METHODS: Sixty-one underweight, stunted children 42 to 60 months of age were recruited in rural Malawi, in southeastern Africa. They received either RTUF or maize and soy flour for 12 weeks. Both supplements provided 2 MJ (500Kcal) of energy daily but had different energy and nutrient densities. Outcome variables were weight and height gain and dietary intake. RESULTS: Before intervention, the mean dietary intake and weight and height gain were similar in the two groups. During the supplementation phase, the consumption of staple food fell among children receiving maize and soy flour but not among those receiving RTUF. There was thus higher intake of energy, fat, iron, and zinc in the RTUF group. Both supplements resulted in modest weight gain, but the effect lasted longer after RTUF supplementation. Height gain was not affected in either group. Periodic 24 hour dietary recalls suggested that the children received only 30% and 43%, respectively, of the supplementary RTUF and maize and soy flour provided. CONCLUSIONS: RTUF is an acceptable alternative to maize and soy flour for dietary supplementation of moderately malnourished children. Approaches aimed at increasing the consumption of supplementary food by the selected recipients are needed. PMID- 14734877 TI - Safety and efficacy of a premixed, rice-based oral rehydration solution. AB - The authors compared the safety and efficacy of a ready-to-use, premixed, rice based oral rehydration solution (R-ORS) with a glucose-based oral rehydration solution (G-ORS), each containing 75 mmol/L sodium, in Mexican children with acute diarrhea for less than 5 days. METHODS: One hundred eighty-nine boys 3 to 24 months old admitted to the hospital with acute diarrhea and signs of dehydration were randomly assigned to receive either G-ORS or R-ORS. Intake and output were measured every 3 hours. RESULTS: In the group treated with R-ORS, significantly fewer patients required supplemental intravenous fluids during the rehydration phase compared to the G-ORS group (1% v 8.7%; P < 0.01). Mean stool output, percent weight gain, ORS intake, urine output, and number of patients who vomited during rehydration were similar in the two groups. The mean total stool output after the first 24 hours of maintenance phase was significantly lower in the R-ORS group than in the G-ORS group. CONCLUSIONS: The authors found rice based ORS to be safe, and its use reduced the rate of intravenous fluid therapy in comparison with the use of a glucose-based ORS. PMID- 14734878 TI - Nutritional status and gastrointestinal structure and function in children with ichthyosis and growth failure. AB - Growth failure occurs in several of the ichthyoses, a heterogeneous group of inherited disorders characterized by thickened or scaly skin. This suggests that there may be common pathogenic mechanisms causing failure to thrive. Previous studies have proposed that a hypermetabolic state induced by epidermal inflammation and hyperproliferation or enteropathy leading to malabsorption and nutritional deficiencies might account for the growth failure in icthyosis. OBJECTIVE: The purpose of this study was to examine the extent of enteropathy and nutritional deficiency in children with severe ichthyosis and growth failure. METHODS: Ten children with different types of ichthyoses and growth failure were studied. RESULTS: Evaluation of gastrointestinal structure and function revealed few abnormalities other than mild fat malabsorption in two patients. Total caloric intake exceeded established requirements for age, height, and weight in all patients. Deficiencies of fat-soluble vitamins were identified in a few subjects. Mild elevations in total calcium and magnesium blood level, elevated hematocrit levels, and constipation were observed in many patients. CONCLUSIONS: Nutritional deficiencies and gastrointestinal abnormalities are uncommon in children with ichthyosis and failure to thrive. Chronic hypovolemia caused by impaired epidermal permeability barrier may be common in this population. PMID- 14734879 TI - Growth and safety in term infants fed reduced-protein formula with added bovine alpha-lactalbumin. AB - OBJECTIVE: To assess growth, tolerance, and biochemical measures of protein status in term infants fed an experimental formula with reduced total protein concentration and enriched in bovine alpha-lactalbumin prepared from an alpha lactalbumin dominant bovine whey. METHODS: Healthy, term, exclusively formula-fed infants < or =14 days postnatal age, between 10th and 90th percentiles in weight and length for age were studied in this randomized, masked, multicenter study. Infants received ad libitum feedings of either experimental or control formula for 12 weeks. Adverse events and acceptability of formulas were assessed every 2 weeks. Weight, length, and head circumference were measured at baseline and every 4 weeks. Serum creatinine, albumin, and blood urea nitrogen were assessed at baseline and study completion. Anthropometric measures were compared to Centers for Disease Control reference ranges using Z scores. RESULTS: One hundred ninety three infants were enrolled. One hundred thirty-four completed the protocol. Seventy-two received experimental formula, and 62 received control formula. The mean baseline body weight was higher in infants fed experimental formula (P = 0.042), so baseline weight was used as a covariate in statistical analyses. There were no differences between groups in gains in weight, length, or head circumference during the study. Mean serum albumin and blood urea nitrogen (BUN) levels were similar at study initiation. At study completion, serum albumin levels were the same in both groups, whereas BUN was significantly higher in infants fed control formula (P = 0.0016). More infants fed control (n = 20) than experimental (n = 15) formula discontinued the study because of adverse events. There were no differences in the adverse event profiles of the groups completing the study. Most of the adverse events were mild and resolved without treatment or sequelae. Acceptability and tolerance of the experimental formula was greater than the control formula, except at 2 weeks. Unacceptable ratings ranged from 0% to 10% (mean, 4.1%) in the experimental formula group and from 1.6% to 14.1% (mean, 7.0%) in the control formula group. CONCLUSIONS: Growth and serum albumin were comparable in infants fed experimental and control formulas for the first 12 weeks of life, suggesting adequate protein nutrition from the alpha-lactalbumin rich formula, despite its lower total protein content. Growth and adverse events data support the safety of the experimental formula. Fewer discontinuations and unsatisfactory ratings among infants fed the experimental formula suggest that it is better tolerated than control formula. PMID- 14734880 TI - Gluten-free diet has a beneficial effect on chromosome instability in lymphocytes of children with coeliac disease. AB - OBJECTIVES: Children with coeliac disease (CD) have an increased number of chromosome aberrations in peripheral blood lymphocytes. Whether genetically determined or a secondary phenomenon in CD, chromosome abnormalities may be involved in the predisposition to cancer in CD patients. The aim of the study was to follow a group of children with CD in whom the initial frequency of chromosome aberrations at diagnosis was known and to measure the same variable after a minimum of 2 years on a gluten-free diet. METHODS: Chromosome aberrations in peripheral blood lymphocytes were determined in 17 patients with CD, before and after at least 24 months of a gluten free diet (mean, 33 months), and in 15 healthy children. The differences in the frequency of aberrations were analyzed by Mann-Whitney U test and Wilcoxon matched-pairs signed-ranks test. RESULTS: Twelve patients adhered to the diet and had a significantly lower frequency of chromosome aberrations than did 5 patients not following the diet (0.16% v 1.2%; P = 0.03), whereas at presentation there had been no difference (1.54% v 1.2%; P = 0.09). The frequency of aberrations at follow-up in patients who were diet adherent was significantly lower than at presentation (1.54% v 0.16%; P = 0.02) and remained unchanged in patients who were not diet adherent (1.2% v 1.2%; P = 1). After at least 24 months of a gluten-free diet, children with CD did not differ from healthy control subjects (0.16% v 0.27%; P = 0.54), whereas children not following the diet had an increased frequency of aberrations (1.2% v 0.27%; P = 0.05). CONCLUSIONS: The frequency of chromosome aberrations in peripheral blood lymphocytes of patients with CD decreased significantly on a gluten-free diet. We conclude that genomic instability is a secondary phenomenon, possibly caused by chronic intestinal inflammation. PMID- 14734881 TI - Symptoms and signs have changed in Swedish children with coeliac disease. AB - OBJECTIVE: To examine symptoms and signs in children with coeliac disease and determine whether the clinical picture at disease onset has changed as incidence of the disease has decreased in the last 10 years. This project was part of the ABIS study (All Babies in Southeast Sweden, born from October 1997 to October 1999). METHODS: Eight paediatric departments in Southeast Sweden recorded all children with coeliac disease and registered symptoms according to a standard form. Data were obtained from 79 children with biopsy-confirmed coeliac disease, 43 contemporary controls, and 65 historic controls. RESULTS: When compared with children with normal intestinal biopsies, children with coeliac disease more often had abdominal distension (odds ratio [OR] = 22.17; 95% confidence interval [CI] OR = 5.00-98.25), thin extremities (OR = 5.89; 95% CI OR = 2.09-16.55), irritability (OR = 6.50; 95% CI OR = 1.83-23.03), and tiredness (OR = 15.43; 95% CI OR = 2.00-119.16). When compared with coeliac children diagnosed at < or =2 years of age in Gothenburg between 1985 and 1989, when the incidence of coeliac disease was three times higher, ABIS patients aged < or =2 years at diagnosis had less often experienced diarrhoea (OR = 0.23; 95% CI OR = 0.12-0.65), suboptimal weight gain (OR = 0.02; 95% CI OR = 0.01-0.10), or suboptimal linear growth (OR = 0.14; 95% CI OR = 0.05-0.39). CONCLUSION: This study indicates that, in parallel to changes in incidence, clinical features of coeliac disease in young children have changed during the last 10 years. PMID- 14734882 TI - Recurrent abdominal pain: symptom subtypes based on the Rome II Criteria for pediatric functional gastrointestinal disorders. AB - OBJECTIVES: Recurrent abdominal pain (RAP) is a common childhood complaint rarely associated with organic disease. Recently, the Pediatric Rome Criteria were developed to standardize the classification of pediatric functional gastrointestinal disorders (FGIDs) using a symptom-based approach. The authors tested the hypothesis that most patients with childhood RAP could be classified into one or more of the symptom subtypes defined by the Pediatric Rome Criteria. METHODS: Using a prospective longitudinal design, new patients with RAP (n = 114) were studied at a tertiary care children's medical center. Before the medical evaluation, parents completed a questionnaire about their child, assessing symptoms defined by the Pediatric Rome Criteria. RESULTS: Of the 107 children for whom medical evaluation revealed no organic etiology for pain, 73% had symptom profiles consistent with the Pediatric Rome Criteria for one of the FGIDs associated with abdominal pain (irritable bowel syndrome, 44.9%; functional dyspepsia,15.9%; functional abdominal pain, 7.5%; abdominal migraine, 4.7%) CONCLUSIONS: This study provides the first systematic empirical evidence that RAP, originally defined by Apley, includes children whose symptoms are consistent with the symptom criteria for several FGIDs defined by the Rome criteria. The pediatric Rome criteria may be useful in clinical research to (1) describe the symptom characteristics of research participants who meet Apley's broad criteria for RAP, and (2) select patients with particular symptom profiles for investigation of potential biologic and psychosocial mechanisms associated with pediatric FGIDs. PMID- 14734883 TI - Prolonged monitoring of esophageal motor function in healthy children. AB - OBJECTIVE: To describe diurnal variations of esophageal motor function in children using combined ambulatory 24-hour esophageal manometry and continuous intraluminal pH measurement (MP24). METHODS: Medical records of all patients referred for the performance of MP24 from 1995 to 2002 at a tertiary care center were reviewed. Patients were selected retrospectively for this study using the following inclusion criteria: (1) no dysphagia, (2) normal upper gastrointestinal barium radiograph, (3) normal esophagogastroduodenoscopy and biopsies, (4) normal stationary esophageal manometry, (5) normal esophageal pH probe, and (6) no gastrointestinal pathology appearing after long-term follow-up. Data from the MP24 of these children were retrospectively analyzed for differences between meal, upright, and supine periods using nonparametric univariate analysis. RESULTS: One hundred twenty-three children had MP24 during the study period. Eleven met the criteria for normality and were included. Their mean age was 12.4 +/- 1.5 years. The number of contractions/minute in the upper, middle, and lower esophageal body differed significantly during meals and in the upright and supine periods (P < 0.01). A significant increase in motor activity occurred during meals (P < 0.01). There was less motor activity observed in the supine position than in the upright position (P < 0.01). There were significant differences in effective (P < 0.05) and total propagated peristalsis among the meal, upright, and supine periods (P < 0.01), with the largest percentage of propagated contractions observed during meals. CONCLUSION: This study provides the first information on prolonged esophageal motor activity in pediatric patients without esophageal disease. MP24 in children demonstrates significant diurnal variations in esophageal motor function that is similar to the findings in studies of healthy adults. PMID- 14734884 TI - Low-dose immunosuppression reduces the incidence of post-transplant lymphoproliferative disease in pediatric liver graft recipients. AB - OBJECTIVES: In pediatric solid organ transplantation, the Epstein-Barr virus (EBV)-related lymphoproliferative disorders (PTLD) still play a major role in post-transplant morbidity and mortality. The aim of the study was to determine the incidence of PTLD in pediatric patients with liver transplant who receive low dose immunosuppression protocols. METHODS: All pediatric patients (n = 269) received a dual immunosuppression therapy consisting of cyclosporine A (initial trough levels, 170-200 microg/L; trough levels for maintenance immunosuppression after 1 year, 80-100 microg/L) and prednisolone (starting dose, 60 mg/m2). Steroids were reduced to 30 mg/m2 after 1 week, followed by a weekly tapering to 5 mg/m2. Seventy-seven of 269 patients were switched to tacrolimus therapy. The authors evaluated the significance of EBV-DNA monitoring by quantitative polymerase chain reaction in identifying patients at risk for PTLD. RESULTS: Patient survival was 90.3%; graft survival was 85.9%. Eight patients lost their grafts because of chronic rejection. The incidence of PTLD was low (0.7%), although a significant EBV viral load was found in 42.4% of the patients. One third of the patients with a viral load of 3,000 genomes/10(5) cells or greater had clinical signs of EBV infection. CONCLUSIONS: The authors conclude that low dose immunosuppressive protocols significantly reduce the incidence of PTLD. In patients treated with that regimen, the monitoring of EBV viral load seems not to be helpful. It can be assumed that low-dose immunosuppression does not suppress EBV-specific cytotoxic CD8+ T cells, thus allowing the host to control EBV infection without the risk of PTLD. Our low-dose immunosuppression protocol did not increase the risk of chronic rejection. PMID- 14734886 TI - Adherent white plaques: an endoscopic finding in eosinophilic esophagitis. PMID- 14734885 TI - Patchy villous atrophy of the duodenum in childhood celiac disease. AB - OBJECTIVES: Patchy villous atrophy of the duodenal mucosa has been described in adults with untreated celiac disease (CD) but not in children. The authors evaluated the presence and the distribution of villous atrophy in children with celiac disease to see whether this histologic pattern exists in children. METHODS: We studied 95 children at diagnosis (Group 1) and seven during gluten challenge (Group 2). We measured anti-endomysium antibodies (EMA) by immunofluorescence on monkey esophagus, antihuman-tissue transglutaminase autoantibodies (anti-tTG Abs) by radioimmunoprecipitation, and HLA-DQ2/DQ8 heterodimers by polymerase chain reaction using specific primers. During upper intestinal endoscopy, at least five duodenal biopsy samples were obtained, one from the duodenal bulb and four from the distal duodenum. RESULTS: Thirteen of 95 (13.7%) patients in Group 1 and in 3 of 7 (42.9%) in Group 2 had patchy villous atrophy of the duodenum. In all 16 patients, villous atrophy of the bulb was present. In four children from Group 1, villous atrophy was observed only in the bulb samples. EMA, anti-tTG Abs, and HLA-DQ2/DQ8 heterodimers were present in all patients. Fourteen of 16 had symptomatic CD, and two were silent, detected during screening in subjects at risk for CD. CONCLUSIONS: This is the first study demonstrating that children with CD may have patchy villous atrophy of the duodenum. The bulb mucosa may be the only duodenal area involved, both at diagnosis and after gluten challenge. Therefore, multiple endoscopic biopsies should always be performed, not only in the distal duodenum, but also in the bulb. PMID- 14734887 TI - Enteritis necroticans with recurrent enterocutaneous fistulae caused by Clostridium perfringens in a child with cyclic neutropenia. PMID- 14734888 TI - Hepatocerebral mitochondrial DNA depletion syndrome: clinical and morphologic features of a nuclear gene mutation. PMID- 14734889 TI - Adolescent herpes simplex viral infection related Ludwig's angina in ulcerative colitis. PMID- 14734890 TI - Vegetarian diet alters the assessment of exocrine pancreatic function with the use of fecal tests. PMID- 14734892 TI - Enzymatic treatment of celiac disease. PMID- 14734891 TI - Food refusal: a sign of nausea? PMID- 14734893 TI - Applying personal reflective critical incident reviews in critical care. AB - Critical care nurses can use the personal reflective critical incident review to subjectively relate common experiences relating to crisis and then devise nursing strategies to help alleviate it. This review article, related to a current situation on an intensive care unit, helps the nurse cope with the stress of the unit. Critical care nurses must react to situations that affect everyone, including themselves. Analyzing their own reactions during a crisis situation helps nurses to accomplish this goal. PMID- 14734894 TI - Living with Diamond Blackfan anemia: a challenge toward survival. AB - As the world of medicine and nursing advance, children with once terminal childhood diseases are now surviving into young adulthood. Nurses must learn to care for pediatric patients who transition to adulthood. This article explores Diamond Blackfan Anemia, once considered a terminal childhood disease that can now be a disease of young adulthood. PMID- 14734895 TI - Taking the "critical" out of critical cardiac care: what does the future hold for cardiac care units? AB - This article identifies the rapid advancement of invasive and noninvasive interventional cardiac technologies. The newest cardiac diagnostics are those awaiting Food and Drug Administration approval will have a significant impact in relation to the acuity of future critically ill cardiac patients. While the elderly population continues to grow, these recipients of healthcare will also have more education available to them and hopefully will be practicing healthier lifestyles. There are several research studies in process that will change the future of critical care patients as well as critical care nurses. This article identifies approaches to the treatments and management of coronary artery disease, heart failure, and cardiac arrhythmias as they impact coronary heart disease, which still remains the leading cause of death in the United States. PMID- 14734896 TI - A new weapon against severe sepsis related to necrotizing fasciitis. AB - Necrotizing fasciitis is a life-threatening infection. The purpose of this article is to review necrotizing fasciitis and nursing care as this disease may progress to sepsis. PMID- 14734897 TI - Mediastinal exploration in the surgical intensive care unit. AB - A continuing education program was developed after a need was identified within the surgical intensive care unit (SICU) for critical care nurses to be able to assist the cardiothoracic surgeon in performing an emergent mediastinal reexploration in the postcardiac surgical patient when an operating room team was unavailable. The program is targeted for SICU nurses experienced in caring for these patients. PMID- 14734898 TI - Accountability and recent developments in nursing: be bold, be holistic: a review of the literature. AB - Contemporary intensive care nurses are confronted daily with accelerated changes and unprecedented challenges. While advancements in critical care and medical technologies may reflect good intentions for patient care, misguided nursing development may pose a sea of fallibility for intensive care nursing. Conceivably, complex technological developments provoke exponential alienation of intensive care nurses from essential nursing duties. Accountability of the nurse is crucial. A winter of discontent is evident as intensive care nurses assume new roles amidst the new technology. Several themes will be presented in this article. PMID- 14734899 TI - Ethical dilemma: voluntarily stopping eating and drinking. AB - Nutritional support is an ethical/legal dilemma that healthcare providers must face when dealing with the elderly population of patients in critical care units. As nurses we have the responsibility to consider the issues concerning this topic. The debate concerning the issue of nutritional support, implications to terminate treatment, and fears concerning liability have shifted the decision making process from the intensive care bedside to the courtroom. Hence, nurses must face this dilemma daily. This article utilizes a case study approach, explores legal issues, discusses ethical guidelines, and identifies techniques for conflict resolution. PMID- 14734900 TI - Hospital nurse staffing and patient outcomes: a review of current literature. AB - An aging nursing workforce, decreased enrollment in nursing schools, financial constraints in healthcare, hospital restructuring and reengineering, and consistent challenges in nursing recruitment and retention have contributed to shortages within the hospital-based nursing workforce. The effects of these shortages have been thought to be associated with various adverse patient outcomes. This article reviews current research studies and presents recommendations for ongoing nursing practice. PMID- 14734901 TI - Women and heart disease. PMID- 14734902 TI - Results of strontium-89 therapy in patients with prostate cancer resistant to chemotherapy. AB - PURPOSE: Strontium-89 (Sr-89) chloride is an effective palliative treatment of the bone metastases of prostate cancer. Chemotherapy has also been shown to have a palliative benefit in this disease. We aimed to determine the benefits and complications of Sr-89 therapy in patients with prostate cancer who had become refractory to chemotherapy. We conducted a retrospective review of 14 treatments administered to 13 patients with chemotherapy-resistant and hormone-resistant prostate cancer. RESULTS: Of the 14 administered treatments, 8 (57%) resulted in improved pain control, with 2 patients able to stop analgesia. The median duration of response was 56 days. No prostate-specific antigen response was seen in the 8 patients tested. There was significant and prolonged bone marrow toxicity, with 6 patients requiring red blood cell transfusion. Prolonged thrombocytopenia was seen, with platelet counts remaining below baseline levels after treatment in all but one patient. Leukopenia was generally mild and not associated with infection. CONCLUSIONS: Sr-89 is an effective treatment of patients with chemotherapy-refractory prostate cancer, but careful and prolonged monitoring of hematologic parameters after therapy is required. PMID- 14734904 TI - FDG PET utility in paraneoplastic sweet syndrome. AB - We describe a 43-year-old woman who presented a sudden onset of fever and migratory arthralgias. Physical examination revealed tender, well-demarcated erythematous papules and plaques, consistent with a Sweet syndrome. After developing systemic symptoms with hepatomegaly, a liver biopsy and FDG PET imaging demonstrated the presence of an aggressive and extended non-Hodgkin T cell lymphoma. This case highlights the usefulness of FDG PET imaging for the screening of this paraneoplastic syndrome. PMID- 14734903 TI - Value of FDG PET imaging in the management of patients with thyroid, neuroendocrine, and neural crest tumors. AB - OBJECTIVES: The purpose of this study was to evaluate the value and impact on management of FDG PET imaging in patients with biopsy-proved neuroendocrine and neural crest tumors, as well as thyroid carcinoma of various types. MATERIALS AND METHODS: This is a retrospective review of imaging and clinical data of 60 consecutive patients presenting for evaluation of suspected recurrence who underwent FDG PET imaging between August 1993 and February 2002. All patients were referred for PET because of equivocal findings on conventional evaluation (most often increasing tumor markers and negative conventional workup) or for restaging. The final diagnosis was established with pathology (n = 19) or at least 6 months of follow-up (n = 41). The FDG images were acquired 1 hour after the intravenous administration of 10 mCi FDG with 1 of 2 dedicated PET tomographs (Siemens ECAT 933, CTI, Knoxville, TN; and GE Advance, General Electric Medical Systems, Milwaukee, WI). RESULTS: Thirty-four patients had recurrent disease and 26 had no evidence of recurrence. FDG PET imaging revealed at least as many focal abnormalities as concurrent CT, magnetic resonance, or other nuclear imaging modalities in 46 of 60 patients (77%). There were 17 patients (28%) in whom FDG PET found abnormalities not seen with other modalities. In the small group (n = 18) of cases of carcinoid, pheochromocytoma, Merkel cell tumor, and neuroblastoma, all cases were true positive (T+) or true negative (T-). In the group of 42 cases of thyroid carcinoma, the sensitivity was 67%. There were 16 T+, 18 T-, and 8 false-negative (F-) cases. Six F- cases presented with increasing thyroglobulin levels and negative whole-body I-131 scans. Four of these 6 F- cases were proved by surgery, 1 by a positive post I-131 therapy scan, and 1 by normalization of thyroglobulin levels after I-131 therapy. Two of the 8 F- PET cases had a positive I-131 scan. FDG PET imaging had an impact on the management of 13 of 60 of these patients (22%) by demonstrating extensive metastases and cancelling surgery (n = 2), and by detecting recurrence and guiding surgery (n = 5) or radiation therapy (n = 6). CONCLUSIONS: FDG PET is helpful in the evaluation of thyroid, neuroendocrine, and neural crest tumors. Although the sensitivity was only 76%, there were no false-positive findings, and FDG PET findings changed the management of 22% of the patients. PMID- 14734905 TI - F-18 fluorodeoxyglucose uptake in atherosclerotic plaque in the mediastinum mimicking malignancy: another potential for error. AB - Atherosclerosis is an inflammatory process accumulating numerous inflammatory cells such as macrophages and lymphocytes. This cellular infiltrate has a high metabolic demand, which can be reflected on F-18 FDG positron emission tomographic (PET) imaging. The FDG uptake in atherosclerotic vessels is usually distinguishable as a result of its linear and continuous appearance and mild level of activity. However, occasionally, atherosclerotic plaques present themselves as focal "hot spots," which can be easily misinterpreted as malignancy. This report emphasizes atherosclerotic plaques as a potential pitfall while interpreting FDG scans. It also highlights the importance of radiographic correlation to avoid such an error. PMID- 14734906 TI - Spontaneous retroperitoneal hemorrhage localized by blood pool scintigraphy. AB - A healthy 17-year-old girl presented to the emergency department with a 1-day history of left upper quadrant abdominal pain associated with nausea and vomiting. Her hematocrit was 22. On physical examination, she had left upper quadrant fullness and tenderness. Initial computed tomography revealed a large, loculated, left-sided retroperitoneal hematoma. Blood pool scintigraphy with labeled red cells revealed a very large photon-deficient area with 3 areas of active bleeding in the upper margin of the cold area. An angiogram showed active extravasation from the left inferior phrenic artery. The patient was felt to have had spontaneous adrenal hemorrhage, likely within a preexisting, large adrenal cyst. Spontaneous hemorrhage into an adrenal cyst is a rare entity that can be life-threatening if not treated early in its course. PMID- 14734907 TI - Unilateral duplicated system: comparative length and function of the kidneys. AB - OBJECTIVE: The objective of this study was to estimate kidney length and function in patients with unilateral duplex kidney. MATERIALS AND METHODS: Thirteen patients with a unilateral duplicated system were reviewed retrospectively. The length of the kidneys was measured by ultrasound, and the relative function of the kidneys was estimated by renal scan. RESULTS: In all patients, the duplex kidney was the left one. The length of the right kidney on the renal ultrasound growth chart was from -1.5 to +0.4 standard deviations from the mean for age, and the left kidney length was from -0.5 to +4.3 standard deviations from the mean. On renal scans the kidneys with a duplicated system contributed 51 to 67% to total renal function; the contralateral ones, 33 to 49%. CONCLUSIONS: Kidneys with a duplicated system may be larger than their counterparts and they may contribute more to total renal function. When a disparity in length between the 2 kidneys is encountered, 1 of the possibilities that should be taken into account is a unilateral duplicated system. PMID- 14734908 TI - F-18 FDG PET: mottled splenomegaly with remission of symptoms after splenectomy in sarcoidosis. PMID- 14734909 TI - Cold splenic lesion on Tc-99m colloid and denatured RBC scintigraphy. PMID- 14734910 TI - Bilateral triceps rhabdomyolysis: an incidental finding on bone scintigraphy. PMID- 14734911 TI - An unusual case of juxtacortical osteosarcoma presenting on Tc-99m MDP bone scan with central photopenia. PMID- 14734912 TI - Chondroblastic osteosarcoma: scintigraphic appearance. PMID- 14734913 TI - Trimetazidine improves myocardial perfusion and left ventricular function in ischemic left ventricular dysfunction. PMID- 14734914 TI - Pericarditis associated with cryptococcal meningitis on Ga-67 scintigraphy. PMID- 14734915 TI - Incidental thallium uptake in bilateral breast lymphoma during myocardial perfusion scintigram. PMID- 14734916 TI - Abnormal FDG PET imaging in tuberculosis appearing like lymphoma. PMID- 14734917 TI - Tc-99m MDP uptake in liver metastases from adenocarcinoma of the duodenum. PMID- 14734918 TI - Ga-67 uptake in thymic carcinoma: utility of Ga-67 scintigraphy in patient management. PMID- 14734919 TI - Atypical carcinoid tumor detected incidentally on Ttc-99m sestamibi myocardial perfusion scintigraphy. PMID- 14734920 TI - Contrast-enhanced magnetic resonance angiography of the carotid arteries: influence of injection rates and volumes on arterial-venous transit time. AB - RATIONALE AND OBJECTIVES: To assess the influence of injection rates and volumes on the arrival time of contrast material (CM) in the common carotid artery, the jugular vein and the resulting arterial-venous transit time. Additionally the relationship between injection parameters and the extent of a CM plateau was evaluated. MATERIALS AND METHODS: In 60 patients a CM injection was necessary to investigate suspected cranial disease. All patients were prospectively assigned to 6 protocol groups with varying volumes of gadolinium dimeglumine (2, 10, 20 mL) and injection rates (0.5, 1, 2, 4 mL/s). Simultaneously to the CM injection, 50 repetitive transverse measurements (1/s) were acquired at the level of the common carotid artery. Based on the resulting signal-time curves, the arrival time of the contrast material in the common carotid artery and the jugular vein, the resulting arterial-venous transit time, the peak enhancement and the extent of a CM-plateau were calculated as a function of the injection parameters. RESULTS: Smaller flow rates (0.5 mL/s) resulted in a longer arrival time in the common carotid artery (mean value 22,6 seconds +/- 2.3) and the jugular vein (mean value 32.6 seconds +/- 2.6) and resulted in longer arterial-venous transit time (mean value 10.1 second +/- 1.9). The volume showed no effect on these parameters. The peak arterial and venous signal intensity and a consistent CM plateau after 50 seconds were dependent on the volume, but not on the injection rate. CONCLUSION: The injection rate showed an influence to the arrival time in the common carotid artery and the jugular vein and also to the arterial-venous transit time. The injected volume only affected the extent of the contrast plateau. A flow rate of 1 to 2 mL/s and a minimum of 20 mL gadolinium dimeglumine are recommended to achieve optimal image quality without venous overlay. PMID- 14734921 TI - Improved accuracy of noninvasive electron beam coronary angiography. AB - OBJECTIVES: We investigated the effect of electrocardiographic (ECG) triggering on the accuracy of coronary electron-beam angiography (EBA) as compared with invasive angiography. METHODS: One hundred thirty-three patients with suspected coronary disease were studied with intravenous coronary EBA and conventional coronary angiography. Patients were divided into 2 groups based upon ECG triggering on the EBA study. Patients were divided into 2 groups based upon different ECG triggering used: 80% R-R interval trigger method (group 1, n = 53) and end-systolic triggering (group 2, n = 80). End-systolic ECG triggering, which started at the end of the T wave in each study, was based on baseline heart rate. RESULTS: Overall sensitivity to detect a > or = 50% luminal stenosis was 69% in group 1 and 91% in group 2 (P = 0.002); specificity was 82% and 94% in group 1 and group 2, respectively (P < 0.001). Using newer triggering techniques (group 2) with EBA, the sensitivity, specificity, and accuracy for patients with disease of the left main coronary artery or 3 vessel disease was 100%, 94%, and 98%, respectively. Nonassessability of coronary segments on 3D-EBA images was reduced from 35% in group 1 to 9% in group 2 patients (P < 0.001). The number of motion free coronary images increased from 67% to 95% from group 1 to group 2 (P < 0.0001). CONCLUSION: End-systolic ECG triggering improves accuracy, image quality, and assessability of segments of coronary EBA for the detection of angiographic coronary artery disease. PMID- 14734922 TI - Potential for differential diagnosis with gadolinium-ethoxybenzyl diethylenetriamine pentaacetic acid-enhanced magnetic resonance imaging in experimental hepatic tumors. AB - RATIONALE AND OBJECTIVES: To investigate whether dynamic and delayed magnetic resonance imaging (MRI) with gadolinium-ethoxybenzyl-diethylenetriamine pentaacetic acid (Gd-EOB-DTPA), a hepatobiliary MRI contrast agent, has potential for the differential diagnosis of experimental hepatic tumors. METHODS: Twelve male rats received N-nitrosomorpholine solution as drinking water to induce hepatic tumors. After injection of Gd-EOB-DTPA, rats were subjected to dynamic and delayed MRI. The relative enhancement (RE) was calculated, and the time of the maximum RE (Tmax) was evaluated. After MRI, liver was histologically analyzed. RESULTS: One hundred sixty-three hepatic tumors 3-12 mm in diameter were induced after 18 weeks of treatment with 0.01 wt/vol% of N nitrosomorpholine, and 81 of them were evaluated. The RE in hyperplastic nodules (HPNs) was significantly higher than that in moderately or poorly differentiated hepatocellular carcinomas (HCCs) in the late phase, whereas there was no significant difference in RE between well-differentiated HCCs and HPNs. The average Tmax in HPNs was about 13 minutes, whereas that of each differentiated HCCs was about 1 minute. CONCLUSIONS: It was possible to differentiate benign HPNs and malignant HCCs (especially well-differentiated HCCs) by evaluating the change of RE or comparison of Tmax with Gd-EOB-DTPA-enhanced MRI. PMID- 14734923 TI - Dual-probe radiofrequency ablation: an in vitro experimental study in bovine liver. AB - OBJECTIVE: To determine whether alternative radiofrequency (RF) application with dual probes has advantages over sequential or simultaneous RF application for creating larger coagulation necrosis. MATERIALS AND METHODS: Using 2 17-gauge internally cooled electrodes and a 200-watt generator (CC-3 model, Radionics), RF energy was applied sequentially (group A, n = 20), simultaneously (group B, n = 20), or alternately (group C, n = 20) to explanted bovine liver. After preliminary experiments with a probe distance of 2-5 cm, a distance of 2 cm and 3 cm were chosen for main experiments. Total duration of the RF application was 10 minutes. In the alternative mode, the energy was applied alternately to both RF electrodes in 2-second intervals. Changes occurring in tissue impedance, current, power output, and temperature at the midpoint between the 2 electrodes were measured. The technical parameters, temperatures, and dimension of the ablated area were compared in the 3 groups using analysis of variance or Kruskal-Wallis test. RESULTS: In the groups B and C, the impedance was gradually decreased during RF application and the mean current flow of both groups was 1550 +/- 130 mA and 1375 +/- 175 mA, respectively. However, in the group A, the impedance was markedly increased during RF application, and the mean current was 940 +/- 484 mA (P < 0.05, between groups A and B, and groups A and C). With 3-cm spacing, the alternative RF application created oval-shaped ablation zones with larger shortest axis diameter at the midpoint than either the sequential or simultaneous RF application: 12.0 +/- 7.1 mm in group A; 27.0 +/- 3.1 mm in group B; 34.9 +/- 3.4 mm in group C (P < 0.05). With 2 cm spacing, the corresponding figures were 25.3 +/- 7.8 mm in group A; 34.9 +/- 7.8 mm in group B; 41.5 +/- 1.8 mm in group C (P < 0.05): the differences between groups A and B, and between groups A and C were statistically significant (P < 0.05). With 3-cm spacing, the temperature at the midpoint between the 2 probes was higher in group C (102 degrees C) than in either the group A (62 degrees C) or the group B (78 degrees C; P < 0.05). With 2 cm spacing, groups B and C showed higher temperature than group A (P < 0.05), but the difference between groups B and C was not significant (P > 0.05). CONCLUSION: The alternative RF application creates larger, more regular ablation zones than either the sequential or simultaneous RF application. This result suggests that use of an alternating RF power application will improve the results of RF ablation for the treatment of larger tumors. PMID- 14734924 TI - Diagnostic performance of a flat-panel detector at low tube voltage in chest radiography: a phantom study. AB - RATIONALE AND OBJECTIVES: To evaluate a large area, cesium iodide amorphous silicon flat-panel detector (CsI/a-Si) at 3 tube voltages to detect simulated interstitial lung disease, nodules, and catheters. METHODS: Simulated interstitial lung disease, nodules, and catheters were superimposed over a chest phantom. Images were generated at 125 kVp, 90 kVp, and 70 kVp at the same surface dose and reduced effective dose equivalent for 90 kVp and 70 kVp and printed on hard copies. Fifty-four thousand observations were analyzed by receiver operating characteristic (ROC). RESULTS: Detectability of linear, miliary, reticular pattern, and nodules over lucent lung as well as of catheters and nodules over obscured chest areas increased at 90 and/or 70 kVp with higher Az values; however, only it was statistically significant for reticular pattern at 70 kVp and nodules at 90 kVp compared with 125 kVp (P < 0.05). The detection of ground glass pattern was worse at lower kVp (P > 0.05). CONCLUSION: For most simulated patterns, differences in diagnostic performance at 70 kVp/90 kVp and 125 kVp were not significant, except for reticular pattern and nodules over lucent lung. PMID- 14734925 TI - Novel echogenic drug-immunoliposomes for drug delivery. AB - RATIONALE AND OBJECTIVES: We have developed novel echogenic immunoliposomes (ELIPs) that can be antibody-conjugated for the specific highlighting of atheroma and atheroma components. The utility of these agents for regional drug delivery has not been evaluated previously. We chose to use an antibiotic as the prototype drug. The concept that an infectious agent may affect the development and progression of atherosclerosis has stimulated trials on the use of antibiotics for coronary syndromes. However, these agents are given systemically with concomitant problems. Development of an agent for local drug delivery may obviate adverse effects and improve treatment efficacy. The aim of this study was to evaluate the potential of our ELIPs for drug incorporation and to demonstrate efficient drug delivery to cultured cells. METHODS: Azithromycin was incorporated into the ELIPs during development. Free drug was removed with a Sephadex G-50 column. Acoustic properties were evaluated using an intravascular ultrasound catheter and quantified by computer-assisted videodensitometry. Human umbilical arterial endothelial cells were infected with Chlamydia pneumoniae. Cells were treated with the drug-ELIP complexes, and infection-forming units were counted using fluorescence techniques. RESULTS: We were able to incorporate a drug into the ELIPs with retention of acoustic properties. The drug-ELIP complex demonstrated effective inhibition of microbial growth in endothelial cells (P < 0.001 vs. empty liposomes and control). CONCLUSIONS: We have developed a novel acoustic drug-liposomal agent that can deliver drugs to cultured cells. Although in vivo translation is required, this technique has potential for site-specific drug delivery. PMID- 14734926 TI - Renal embolization: feasibility of magnetic resonance-guidance using active catheter tracking and intraarterial magnetic resonance angiography. AB - RATIONALE AND OBJECTIVES: Magnetic resonance (MR)-guidance of endovascular interventions offers various advantages, including the absence of ionizing radiation, excellent soft tissue contrast, and multiplanar and functional imaging capabilities. The objective of this study was to assess the feasibility of MR guided renal embolization using active catheter tracking with automatic slice positioning and intraarterial contrast-enhanced MR angiography (MRA). MATERIALS AND METHODS: MR-guided embolization of 16 kidneys was attempted in 15 pigs using real-time tracking of active 5-Fr. catheters. Embolization was monitored by selective intraarterial projection MRA. Intraarterial three-dimensional (3D) MRA was used for the assessment of embolization results. Additional pathologic correlation was available in 2 animals. The image quality of intraarterial 3D contrast-enhanced-MRA was rated by an independent radiologist who was not involved in the animal experiments. RESULTS: Active catheter tracking with automatic slice positioning allowed reliable catheter guidance and catheterization of the renal artery in all animals. Embolization was successful in all kidneys (11 left, 5 right), as verified by intraarterial 3D contrast enhanced MRA (ce-MRA) and/or pathology. The image quality of intraarterial 3D ce MRA was rated excellent in 10 animals, moderate in 4 animals, and poor in 1 animal. CONCLUSION: Renal embolization using active catheter tracking and intraarterial ce-MRA is feasible. Selective intraarterial ce-MRA allows the assessment of blood supply and organ perfusion before, during, and after therapeutic interventions, thereby complementing MR-guided endovascular interventions. PMID- 14734927 TI - The influence of pulse and respiration on spinal cerebrospinal fluid pulsation. AB - RATIONALE AND OBJECTIVES: The aim of the study is to elucidate the location and amount of spinal cerebrospinal fluid pulsations and to differentiate and quantify the cardiac and the respiratory influence. MATERIALS AND METHODS: An echo planar imaging sequence was applied to 5 different levels of the spinal canal of 7 healthy volunteers. The amount of maximal flow and respiratory signal variation were determined by a time and frequency domain analysis, respectively. RESULTS: CSF pulsation was high in the anterior cervical and in the thoracolumbar spine. Respiratory influence rose by 19% at C1 and by 28% at T12. The systolic flow was elevated during late expiration and the diastolic upward movement was pronounced by early expiration. CONCLUSION: The pulsation in the lower spine seems to be related to a second motor of CSF movement because there is a rising respiratory influence and a reappearance of pulsation waves. Physiological spinal CSF pulsation contains a relevant respiratory component. PMID- 14734929 TI - Epileptic encephalopathies: a brief overview. AB - Epileptic encephalopathies are conditions in which neurologic deterioration is attributable entirely or partly to epileptic activity. It can be due to very frequent or severe seizures and/or to subcontinuous paroxysmal interictal activity. The former mainly consists of Dravet syndrome, in which patients have seizures from the middle of the first year of life and repeat episodes of severe febrile status epilepticus and migrating partial epilepsy in infancy, in which from the first trimester of life, partial seizures affect various areas of the cortex randomly and in a subcontinuous fashion. In Rasmussen syndrome, also, epileptic activity contributes at least partly to the neurologic deterioration. Subcontinuous paroxysmal interictal activity affects newborn infants with suppression bursts, thus consisting in either Ohtahara syndrome or neonatal myoclonic encephalopathy. In infants, it is either myoclonic epilepsy of nonprogressive encephalopathy or West syndrome. In school-age children, it consists of various types of generalized seizures combined with slow spike waves of the Lennox-Gastaut syndrome, myoclonic-astatic epilepsy, and continuous spike waves in slow sleep combined with various motor or cognitive deficits including negative myoclonus, orofacial dyspraxia, Landau-Kleffner syndrome, and frontal lobe syndrome. Treatment differs for all of these syndromes. It is important to avoid potential drug-induced worsening, and valproate is preferred when a definitive diagnosis is not reached in children and especially infants. PMID- 14734930 TI - Epileptic encephalopathies in early infancy with suppression-burst. AB - Early infantile epileptic encephalopathy with suppression-burst, or Ohtahara syndrome (OS), and early myoclonic encephalopathy (EME) are epileptic encephalopathies with onset of frequent seizures in the neonatal and early infancy period and with a characteristic EEG pattern, namely, suppression-burst, in which higher-voltage bursts of slow waves mixed with multifocal spikes alternate with isoelectric suppression phase. Their nosologic independence is now widely accepted, although some controversy initially occurred because of their common characteristics such as age of onset, EEG features, seizure intractability, and poor prognosis. Major differences between the two syndromes include (1) tonic spasms in OS versus partial seizures and erratic myoclonias in EME, (2) continuous suppression-burst pattern in both waking and sleeping states in OS versus this EEG pattern almost limited to sleep in EME, and (3) static structural brain damage in OS versus genetic or metabolic disorders in EME. The most important differentiating point is their evolutional pattern with age, which may reflect their pathophysiologic difference. Ohtahara syndrome evolves to West syndrome and further to Lennox-Gastaut syndrome with age, but EME demonstrates no unique evolution; namely, it continues as such for a long time or changes into partial epilepsy or severe epilepsy with multiple independent spike foci. PMID- 14734931 TI - Infantile epileptic encephalopathy with hypsarrhythmia (infantile spasms/West syndrome). AB - Infantile spasms is a unique disorder peculiar to infancy and early childhood. In this article, the clinical manifestations and electroencephalographic features of the disorder are described. The possible pathophysiologic mechanisms underlying infantile spasms and the relation of this disorder to other childhood encephalopathies are discussed. Finally, the treatment of patients with infantile spasms and their long-term outcome are briefly reviewed. PMID- 14734932 TI - Lennox-Gastaut syndrome (childhood epileptic encephalopathy). AB - Lennox-Gastaut syndrome (LGS) is a childhood epileptic encephalopathy characterized by an electroclinical triad of generalized slow spike wave (SSW) activity in the EEG, multiple types of epileptic seizures, and slow mental development. It is usually subdivided into symptomatic and cryptogenic types, the latter accounting for at least one fourth of all patients. Symptomatic cases are due to diverse cerebral conditions, which are usually bilateral, diffuse, or multifocal, involving cerebral gray matter. Twenty percent of all patients with LGS have prior infantile spasms with hypsarrythmia. The characteristic interictal EEG pattern of LGS is 1.5 to 2.5 Hz SSW activity, which is bilaterally synchronous, dominant over the frontocentral regions, and usually symmetric. There are varying degrees of slowing of the background. Sleep discloses paroxysms of generalized fast (10 to 25 Hz) rhythmic activity. PMID- 14734933 TI - Severe epilepsy with multiple independent spike foci. AB - SUMMARY: Severe epilepsy with multiple independent spike foci is an electroclinical entity with the following characteristics: (1) EEG showing multiple independent spike foci (three or more foci in both hemisphere, i.e., at least one in each hemisphere) and diffuse slowing of the background activity, (2) very frequent multiple types of seizures but mainly generalized minor seizures, (3) frequent association with mental retardation and neurologic abnormalities, (4) underlying causes of various nonspecific prenatal, perinatal, and postnatal cerebral conditions, and (5) poor prognoses for seizures and psychomotor development. It represents a diffuse encephalopathy with mutual transition between other age-dependent epileptic encephalopathies. This electroclinical condition may be appropriately named Markand-Blume-Ohtahara syndrome, after Markand and Blume, who initially described in detail the EEG pattern of multifocal spike discharges with its major clinical correlates, and Ohtahara, who extensively studied the group of patients with both clinical and EEG characteristics and proposed the concept of severe epilepsy with multiple independent spike foci, classifying it as one type of generalized epilepsy. PMID- 14734934 TI - Epileptic encephalopathies with myoclonic seizures in infants and children (severe myoclonic epilepsy and myoclonic-astatic epilepsy). AB - Myoclonic attacks are not characteristic of a specific syndrome. In infancy and early childhood, they are often observed in the context of syndromes that are associated with other types of seizures and with cognitive impairment but no obvious brain lesion. Characterization of the associated seizures and age of expression allows inclusion of a number of cases in two main subgroups: severe myoclonic epilepsy (SME, or Dravet syndrome) and myoclonic-astatic epilepsy (MAE). Severe myoclonic epilepsy is an epileptic encephalopathy with invariably poor outcome in which myoclonic seizures, though frequently observed, may be absent altogether in some children. Prolonged and repeated febrile and afebrile convulsive seizures starting in infancy are the main feature and are probably causally related to cognitive decline. One third of children harbor mutation of the SCN1A gene, but the genetics of SME is probably more complex than expected with simple monogenic disorders. Treatment is usually disappointing. Myoclonic astatic epilepsy is perhaps more a conceptual category of idiopathic myoclonic epilepsy than a discrete syndrome. Childhood-onset myoclonic-astatic attacks are the characteristic seizures associated in most with episodes of nonconvulsive status and generalized tonic-clonic seizures. Outcome is unpredictable. Either remission within a few years with normal cognition or long-lasting intractability with cognitive impairment is possible. Likewise, the effectiveness of antiepileptic drugs is variable. A number of cases of myoclonic epilepsies in infancy and early childhood, however, remain unclassified, and intermediate forms between the different syndromes exist. They must be distinguished from other syndromes with frequent brief attacks and repeated falls, especially the Lennox Gastaut syndrome. This differentiation is often difficult and may require extensive neurophysiologic studies. PMID- 14734935 TI - Epileptic encephalopathy of late childhood: Landau-Kleffner syndrome and the syndrome of continuous spikes and waves during slow-wave sleep. AB - Landau-Kleffner syndrome (LKS) and the syndrome of continuous spikes and waves during slow wave sleep (CSWS) are two points on the spectrum of functional childhood epileptic encephalopathies. They are characterized by a severe paroxysmal EEG disturbance that may permanently alter the critical synaptogenesis by strengthening synaptic contacts that should have been naturally "pruned." The much more common benign epilepsy with centrotemporal spikes is also related to LKS and CSWS by a common pathophysiology. Although prognosis in LKS and CSWS for seizure control is good, cognitive function declines and permanent neuropsychologic dysfunction is seen in many cases. This permanent damage is most evident in those patients who had early-onset EEG abnormality and a prolonged active phase of continuous spike-and-wave discharges during sleep. If the active phase of paroxysmal activity persists for over 2 to 3 years, even successful treatment does not resolve neuropsychologic sequelae. In LKS, the paroxysmal activity permanently affects the posterior temporal area and results in auditory agnosia and language deficits; in CSWS, the frontal lobes are more involved and other cognitive disturbances predominate. Aggressive treatment should include high-dose antiepileptic drugs, corticosteroids, and surgery in specific cases. PMID- 14734937 TI - Framing the problem of measuring and improving healthcare quality: has the Quality Health Outcomes Model been useful? AB - OBJECTIVE: The objective of this study was to determine the uses of the Quality Health Outcomes framework and indicator categories in the healthcare literature. DATA SOURCES: We studied personal communications and conducted a literature search using computerized databases since 1997, when the recommendations of the Invitational Conference on Measures and Outcomes of Care Delivery were available. PRINCIPAL FINDINGS: The Quality Health Outcomes Model has been used explicitly to frame a small number of research summaries and programs. The outcome indicator categories can be found in several "report card" initiatives in the United States and Canada. Use of these outcome categories, thought to be sensitive to nursing care inputs, has grown since 1977, with a rising number of uses linked to system or organizational factors or interventions. CONCLUSIONS: This model and others like it are increasingly forming the conceptual framework for studies that evaluate quality and system interventions to improve care. However, the available data continue to require the linking of negative outcomes (adverse events, complications) to structural and process inputs that reflect nursing care. An urgent need remains to incorporate this broader range of outcomes into available databases. PMID- 14734938 TI - Quality of evidence: usefulness in measuring the quality of health care. AB - BACKGROUND: In 1998, the Committee on Health Care Quality in America was created and given the charge of devising a strategy to achieve substantial improvement in the quality of health care for all Americans. One strategy to move the quality agenda forward is the use of evidence by both providers of health care and consumers of health care. One feature of this agenda, evidence-based practice, refers to a hierarchy of evidence ranging from individual randomized, controlled trials to expert opinion. OBJECTIVES: The purposes of this article are to describe the evidence base in nursing, discuss the quality and strength of nursing's evidence, illustrate the application of the Quality Health Outcomes Model, and present recommendations for practice, research, and policy to increase nursing's contribution to quality health care. RESULTS AND RECOMMENDATIONS: Nurses everywhere must use innovative solutions to operationalize the "evidence" in evidence-based nursing. The Quality Health Outcomes Model (QHOM) provides a useful way of advancing research and evidence about the quality of health care in America. In concert with the conceptual framework for the National Health Care Quality Report, the QHOM provides a map for identifying evidence gaps and research questions arising from the model and conceptual framework, as well as evidence synthesis (integrating methodologic quality) driven by theoretical understanding. PMID- 14734940 TI - Cross-sectional versus longitudinal performance assessments in the management of diabetes. AB - BACKGROUND: Performance assessments help to quantify the level of adherence with practice standards and are often used to measure and compare the quality of care. However, most performance assessments are based on a cross-sectional analysis of patient information, whereas patient care is inherently longitudinal. This discordance could confound the relationship between the performance measure and the true quality of care. OBJECTIVE: The objective of this study was to illustrate differences in performance assessment as measured by a traditional cross-sectional analysis compared with a longitudinal analysis. METHODS: We conducted a cross-sectional and longitudinal analysis of a cohort of diabetic patients in an integrated delivery system having primary care visits and hemoglobin A1c (HBA1c) testing in both 1999 and 2000. RESULTS: In the cross sectional analysis of 4661 patients, we found a modestly increasing proportion achieved an HBA1c level of <8.0%: 73.1% in 1999 and 75.6% in 2000. Longitudinal analysis, however, suggested that certain subsets of patients were more likely to switch from good to poor control or retain their level of poor control over the 2 years studied. In particular, compared with whites, blacks were 1.76 (95% confidence interval [CI], 1.31-2.37) times as likely to switch from good to poor control and only 0.56 (95% CI, 0.41-0.76) times as likely to switch from poor to good control. Patients aged 35 to 49 were 2.54 (95% CI, 1.79-3.45) times as likely to switch from good to poor and only 0.66 (95% CI, 0.47-0.94) times as likely to switch from poor to good control than patients over age 64 years. CONCLUSIONS: Cross-sectional performance assessments could mask changes in diabetes control among individuals belonging to a cohort and, conceptually, are poorer indicators of care process than longitudinal measures. In addition, longitudinal analyses suggest the influence of patient sociodemographic factors on the performance assessment that should be accounted for when comparing quality of care for diabetes. PMID- 14734939 TI - Evaluating the role of patient sample definitions for quality indicators sensitive to nurse staffing patterns. AB - BACKGROUND: Administrative data are an attractive data source for the construction of quality indicators to assess and monitor quality of nursing care in hospitals. Current approaches to constructing measures from discharge abstracts apply substantial restrictions to exclude patients at high risk or with preexisting conditions. This study evaluates whether broader sample definitions combined with risk adjustment would allow for larger samples and increase analytic power. METHODS: Eight indicators were constructed from discharge abstracts of major surgical and medical patients from 799 hospitals in 11 states using existing definitions: pneumonia, urinary tract infection, decubitus ulcers, central nervous system complications, shock, sepsis, pulmonary failure, and upper gastrointestinal bleeding. We tested the effect of broadening the samples in 4 ways: comparing indicator rates in the broader and restrictive samples; assessing correlations of hospital ranks in the broader and restrictive samples; performing clinical reviews of cases in the added samples; and using different samples in regressions of indicators on nurse staffing variables, adjusting for patient risk. RESULTS: Indicator rates in the broader samples tended to be higher but did not change hospital rankings significantly. Clinical review suggested that many sample restrictions could be dropped. Using indicators based on broader definitions, coefficients on staffing variables increased in magnitude. CONCLUSION: Less restrictive sample definitions were shown to be feasible and increased the sensitivity of the indicators and thus the power of the analysis. Particularly in surgical patients, the samples could be broadened, although more conservative definitions appeared appropriate for medical patients. PMID- 14734941 TI - Nursing process outcome linkage research: issues, current status, and health policy implications. AB - BACKGROUND: The use of large clinical datasets to assess the effectiveness of health care is of growing interest in continuing efforts to understand the impact of healthcare costs on quality. Correspondingly, there is a greater need to define and measure outcomes that are sensitive to nursing interventions. However, concerns exist about the ability to amass and use large clinical nursing datasets to assess the effectiveness of nursing interventions. Some nursing studies have used large clinical datasets to examine patterns of nursing diagnoses, interventions, and outcomes. Among patient populations, however, systematic effectiveness studies of nursing process and outcome linkages at the individual nurse and patient level of analysis are essentially nonexistent. This is largely the result of slow development of nursing classifications, reference terminologies, and reference information standards. Nursing information systems have an unprecedented potential for documentation of nursing practice, as well as the accumulation and analysis of large clinical datasets, to improve nursing performance, increase nursing knowledge, and provide data and information necessary for nursing to participate in the formulation of healthcare policy. OBJECTIVES: A literature search shows that a common framework is beginning to evolve that represents nursing's essential information, eg, the Nursing Minimum Data Set, Management Minimum Data Set, and several standardized nursing languages. Extensive research and other initiatives have produced 1) nursing languages and reference terminologies that span healthcare settings; 2) information models; and 3) standards for datasets supporting information systems. A number of issues remain, however, that concern the development of uniform nursing datasets, definitions of outcomes, quality of nursing data, information system design, and methods of data analysis. We review nursing process outcome research, clarify issues inherent in nursing effectiveness research, and discuss implications for nursing and health policy. PMID- 14734942 TI - Promoting patient safety and enabling evidence-based practice through informatics. AB - OBJECTIVES: The purposes of this article are to highlight the role of informatics in promoting patient safety and enabling evidence-based practice (EBP), 2 significant aspects for assuring healthcare quality; to delineate some challenges for the future; and to provide key recommendations for education, practice, policy, and research. METHODS: First, we describe the components of an informatics infrastructure for patient safety and evidence-based practice. Second, we address the role of informatics in 4 areas: 1) information access; 2) automated surveillance for real-time error detection and prevention; 3) communication among members of the healthcare team; and 4) standardization of practice patterns. Last, we delineate some future challenges for nursing and for informatics and provide key recommendations for education, practice, policy, and research. RESULTS: The components of an informatics infrastructure are available and applications that bring together these components to promote patient safety and enable EBP have demonstrated positive or promising results. CONCLUSIONS: Challenges must be addressed so that an informatics infrastructure and related applications that promote patient safety and enable EBP can be realized. PMID- 14734943 TI - Nurse burnout and patient satisfaction. AB - BACKGROUND: Amid a national nurse shortage, there is growing concern that high levels of nurse burnout could adversely affect patient outcomes. OBJECTIVES: This study examines the effect of the nurse work environment on nurse burnout, and the effects of the nurse work environment and nurse burnout on patients' satisfaction with their nursing care. RESEARCH DESIGN/SUBJECTS: We conducted cross-sectional surveys of nurses (N=820) and patients (N=621) from 40 units in 20 urban hospitals across the United States. MEASURES: Nurse surveys included measures of nurses' practice environments derived from the revised Nursing Work Index (NWI-R) and nurse outcomes measured by the Maslach Burnout Inventory (MBI) and intentions to leave. Patients were interviewed about their satisfaction with nursing care using the La Monica-Oberst Patient Satisfaction Scale (LOPSS). RESULTS: Patients cared for on units that nurses characterized as having adequate staff, good administrative support for nursing care, and good relations between doctors and nurses were more than twice likely as other patients to report high satisfaction with their care, and their nurses reported significantly lower burnout. The overall level of nurse burnout on hospital units also affected patient satisfaction. CONCLUSIONS: Improvements in nurses' work environments in hospitals have the potential to simultaneously reduce nurses' high levels of job burnout and risk of turnover and increase patients' satisfaction with their care. PMID- 14734944 TI - Is more better?: the relationship between nurse staffing and the quality of nursing care in hospitals. AB - OBJECTIVE: The objective of this study was to examine the effects of nurse staffing and process of nursing care indicators on assessments of the quality of nursing care. RESEARCH DESIGN: This study examined the variation in inpatient hospital staff nurses' assessments of the quality of nursing care and the effects of nurse staffing (patient workload), patient safety problems (medication errors and patient falls with injuries), and unfinished care (number of nursing tasks left undone) on the variation in those assessments. Secondary analysis of a survey of nurses using multivariate regression models was undertaken. SUBJECTS: Data were derived from a 1999 statewide survey of 8670 inpatient staff nurses working in acute care hospitals in Pennsylvania. RESULTS: Quality of nursing care ratings were significantly associated with the number of patients who nurses care for, rates of unfinished care for those patients, and the frequency of patient safety problems. The effect of patient workload on quality ratings was attenuated substantially by the effects of unfinished care and patient safety problems. Unfinished care had the strongest relationship of all, with over 40% of the variation in quality ratings associated with the number of tasks left undone. CONCLUSIONS: Assessments of the quality of nursing are associated with both structural (workload) and process of care indicators (unfinished clinical care and patient safety problems), with the relationship strongest between process of care and quality. Explicating the interrelationship between structure and process of care is key to understanding the influence of both on quality. Studies that assess the causal influence of these features on quality of care and patient outcomes are warranted. PMID- 14734945 TI - Good diabetes care: more than cheap meds. PMID- 14734946 TI - Health insurance status, cost-related medication underuse, and outcomes among diabetes patients in three systems of care. AB - BACKGROUND: Chronically ill patients often experience difficulty paying for their medications and, as a result, use less than prescribed. OBJECTIVES: The objectives of this study were to determine the relationship between patients with diabetes' health insurance coverage and cost-related medication underuse, the association between cost-related underuse and health outcomes, and the role of comorbidity in this process. RESEARCH DESIGN: We used a patient survey with linkage to insurance information and hemoglobin A1C (A1C) test results. PATIENTS: We studied 766 adults with diabetes recruited from 3 Veterans Affairs (VA), 1 county, and 1 university healthcare system. MAIN OUTCOMES: Main outcomes consisted of self-reported medication underuse as a result of cost, A1C levels, symptom burden, and Medical Outcomes Study 12-Item Short-Form physical and mental functioning scores. RESULTS: Fewer VA patients reported cost-related medication underuse (9%) than patients with private insurance (18%), Medicare (25%), Medicaid (31%), or no health insurance (40%; P <0.0001). Underuse was substantially more common among patients with multiple comorbid chronic illnesses, except those who used VA care. The risk of cost-related underuse for patients with 3+ comorbidities was 2.8 times as high among privately insured patients as VA patients (95% confidence interval, 1.2-6.5), and 4.3 to 8.3 times as high among patients with Medicare, Medicaid, or no insurance. Individuals reporting cost-related medication underuse had A1C levels that were substantially higher than other patients (P <0.0001), more symptoms, and poorer physical and mental functioning (all P <0.05). CONCLUSIONS: Many patients with diabetes use less of their medication than prescribed because of the cost, and those reporting cost-related adherence problems have poorer health. Cost-related adherence problems are especially common among patients with diabetes with comorbid diseases, although the VA's drug coverage may protect patients from this increased risk. PMID- 14734947 TI - Missed appointments and poor glycemic control: an opportunity to identify high risk diabetic patients. AB - OBJECTIVE: When patients miss scheduled medical appointments, continuity and effectiveness of healthcare delivery is reduced, appropriate monitoring of health status lapses, and the cost of health services increases. We evaluated the relationship between missed appointments and glycemic control (glycosylated hemoglobin or HbA1c) in a large, managed care population of diabetic patients. RESEARCH DESIGN AND METHODS: Missed appointment rate was related cross sectionally to glycemic control among 84,040 members of the Kaiser Permanente Northern California Diabetes Registry during 2000. Adjusted least-square mean estimates of HbA1c were derived by level of appointment keeping (none missed, 1 30% missed, and >30% missed appointments for the calendar year) stratified by diabetes therapy. RESULTS: Twelve percent of the subjects missed more than 30% of scheduled appointments during 2000. Greater rates of missed appointments were associated with significantly poorer glycemic control after adjusting for demographic factors (age, sex), clinical status, and health care utilization. The adjusted mean HbA1c among members who missed >30% of scheduled appointments was 0.70 to 0.79 points higher (P <0.0001) relative to those attending all appointments. Patients who missed more than 30% of their appointments were less likely to practice daily self-monitoring of blood glucose and to have poor oral medication refill adherence. CONCLUSION: Patients who underuse care lack recorded information needed to determine level of risk. Frequently missed appointments were associated with poorer glycemic control and suboptimal diabetes self management practice, are readily ascertained in clinical settings, and therefore could have clinical utility as a risk-stratifying criterion indicating the need for targeted case management. PMID- 14734948 TI - Decreasing variation in the use of hospice among older adults with breast, colorectal, lung, and prostate cancer. AB - BACKGROUND: Utilization of hospice services has been shown to vary by place of residence and patient characteristics. OBJECTIVES: The purpose of this study was to examine whether such variation has changed over time. Hospice utilization is examined as a function of sociodemographic characteristics, geographic location, type of insurance, and year of death. RESEARCH DESIGN: This study used a retrospective cohort design. SUBJECTS: We used data from the linked Surveillance, Epidemiology and End Results (SEER)-Medicare database to study hospice utilization in subjects aged 67 and older diagnosed with breast, colorectal, lung, or prostate cancer from 1991 to 1996 and who died between 1991 and 1999. RESULTS: Of the 170,136 subjects aged 67 and older who died from 1991 through 1999, 51,345 (30.2%) were enrolled in hospice before they died. Hospice utilization varied significantly by patient characteristics, including type of insurance, age, marital status, race and ethnicity, gender, urban versus rural residence, type of cancer, income level, and education level. This variation, however, decreased over time for subgroups defined by type of insurance, marital status, urban residence, and income. Variation in hospice use increased over time as a function of age and type of cancer. There was no change in variation in use in blacks compared with non-Hispanic whites over time. CONCLUSIONS: The variation in hospice use by several patient characteristics is decreasing over time, a finding consistent with the manner in which new medical technologies diffuse. PMID- 14734949 TI - Predicting resource utilization in a Veterans Health Administration primary care population: comparison of methods based on diagnoses and medications. AB - BACKGROUND: Valid methods of predicting resource utilization in primary care populations are needed. We compared the predictive validity of a method based on diagnoses from administrative data (Adjusted Clinical Groups [ACGs]) and a method using medication profiles (Chronic Disease Index [CDI]). METHODS: This retrospective cohort study included 31,212 primary care patients in a Veterans Health Administration (VA) network who received outpatient medication prescriptions in 1999 and who had VA utilization in 1999 and 2000. ACG and CDI classifications were determined using 1999 data. Analyses compared the predictive validity with respect to outpatient clinic visits and days of hospital care. RESULTS: Both ACGs and CDI explained a higher proportion of the variance in outpatient visits than demographic data alone. However, explained variance was higher for ACGs. For example, ACGs explained 30.2% of the variance in total visits in 1999, compared with 8.8% for the CDI. Results were similar for 2000, although the explained variance declined for both methods (eg, 16.3% and 5.7%, respectively, for total visits). Results were similar in analyses examining the discrimination of the 2 methods to predict hospital use; for example, c statistics for ACGs and CDI scores were 0.86 versus 0.70, respectively (P <0.05), for 1999 and 0.72 and 0.65, respectively (P <0.05), for 2000. CONCLUSION: Among VA patients, ACGs had superior predictive validity than the CDI, a newer nonproprietary method based on pharmacy data. The findings suggest that diagnosis based measures could be preferable for ambulatory case-mix adjustment and are valid across a wide range of populations. PMID- 14734950 TI - Pain and the use of outpatient services among persons with HIV: results from a nationally representative survey. AB - BACKGROUND: The negative impact of pain on health-related quality of life has been documented for persons with human immunodeficiency virus (HIV). Furthermore, pain could be an important factor in seeking medical care. However, the relationship between pain and health service utilization is poorly understood. OBJECTIVES: The objective of this study was to investigate the effect of pain on use of outpatient services among a nationally representative sample of adults receiving medical care for HIV using Andersen's Behavioral Model of Health Services Use. METHODS: We used Poisson regression to assess outpatient use over 6 months among 2267 respondents in the HIV Cost and Services Utilization Study. Key predisposing variables include gender and race/ethnicity. Enabling factors include income and insurance. Need factors include pain, CD4 count, and diagnosis of acquired immunodeficiency syndrome (AIDS). RESULTS: Sixty-seven percent of respondents reported experiencing pain during the previous 4 weeks. Self-reported pain was higher among those with AIDS, intravenous drug-using females, the unemployed, and those without a baccalaureate degree, but lower among blacks. Patients reporting more pain and those developing more pain used more outpatient services. Poorer health (CD4 count <50 cells/mm3, less energy) was also associated with higher use. Persons with Medicare, Medicaid, or private insurance were more likely to have used outpatient services than the uninsured. Persons with a private HMO were no more likely to use services than those without insurance. CONCLUSIONS: Improved pain management could reduce outpatient use for persons experiencing pain and lead to substantial cost savings. Inequalities in outpatient use related to insurance are evident. PMID- 14734951 TI - Outcome expectancies, functional outcomes, and expectancy fulfillment for patients with shoulder problems. AB - OBJECTIVES: This study was conducted to evaluate the relationship among patient outcome expectancies, perceived shoulder function changes, and perceptions of expectancy fulfillment. METHODS: Patients (n = 199) treated for shoulder problems at one orthopedic surgeon's office completed a baseline survey comprised of measures of outcome expectancies, shoulder function, health status, and demographics. At 1 month, 2 months, and 3 months, patients completed a mailed follow-up survey comprised of all baseline measures except demographic variables. At 3 months, a measure of expectancy fulfillment was added. A general linear modeling approach was used to assess the significance and effect size of 1) outcome expectancies on changes in shoulder function; and (2) outcome expectancies, shoulder function changes, and their interaction on perceptions of expectancy fulfillment. RESULTS: Outcome expectancies significantly predicted changes in shoulder function and accounted for 10% of the variance in functional improvement. The improvement difference between patients with high expectancies compared with those with low expectancies was clinically relevant (4.57 points), as it was greater than the minimal clinically important difference (3.02 points). Outcome expectancies and shoulder function changes significantly predicted patients' perceptions of fulfilled expectancies, but their interaction was not statistically significant. CONCLUSIONS: Results highlight the importance of patient expectancy in medical encounters. The findings suggest the need for interventions targeting patient expectancies such as including discussions about expectancy in patient-physician negotiations of therapeutic plans. Results also underscore the need for the development of better measures of outcome expectancies and expectancy fulfillment. PMID- 14734952 TI - Outcome of patients with cardiac diseases admitted to coronary care units: a report from Lazio, Italy. AB - BACKGROUND: Coronary care units (CCUs) currently treat a variety of diseases, but little is known about the effectiveness of CCUs on heart conditions other than acute myocardial infarction. OBJECTIVES: The objectives of this study were to evaluate the association between direct admission to CCUs and the risk of inhospital death in patients with heart disease, to investigate factors affecting direct admission to a CCU, and to assess the effect of CCU admission on the use of invasive procedures in patients with arrhythmias. RESEARCH DESIGN: We conducted a retrospective analysis of discharge-abstract data from Lazio, Italy, hospitals. We used logistic regression, propensity score, and instrumental variable analysis to compare inhospital risk of death between patients admitted to CCUs and to ordinary wards in 13 different groups of heart disease. We used linear regression to study the association between the rate of CCU admission and the relative risk of death. RESULTS: The study included 181,049 heart disease admissions, of which 8620 were admitted to CCUs (4.8%). Risk of death was significantly lower in patients admitted directly to CCUs for "acute myocardial infarction" (odds ratio [OR], 0.57), "acute ischemic heart disease" (OR, 0.55), and "other arrhythmias" (OR, 0.56). Mortality ORs were inversely related to the rate of CCU admission. CCU patients with arrhythmias received more invasive procedures (OR, 2.70) than non-CCU patients. CONCLUSION: Direct admission to a CCU is associated with a decrease in mortality for patients with "acute myocardial infarction," "acute heart ischemia," and "other arrhythmias." Patients most likely to benefit from CCU care are preferentially admitted to CCUs. CCUs make larger use of invasive procedures than ordinary wards. PMID- 14734953 TI - Evaluating the planned substitution of the minimum data set-post acute care for use in the rehabilitation hospital prospective payment system. AB - OBJECTIVE: The objective of this study was to evaluate the payment implications of substituting the Minimum Data Set-Post Acute Care (MDS-PAC) for the FIM trade mark instrument for use in the planned prospective payment system (PPS) for inpatient rehabilitation hospitals. FIM trade mark is a trademark of the Uniform Data System for Medical Rehabilitation, a division of UB Foundation Activites, Inc. RESEARCH DESIGN: We used a prospective cross-sectional design using consecutive sampling. SUBJECTS: We studied all Medicare admissions with stays of 3 days or more over a 2-month period to 50 inpatient rehabilitation hospitals in 22 states. MEASUREMENTS AND METHODS: Each participating institution completed both the FIM and the MDS-PAC assessments on all participants. Items from the MDS PAC were combined and translated to create "FIM-like" items. We assessed agreement of classification into prospective payment cells using FIM assessment data and also using MDS-PAC data. Statistical adjustments were applied to improve the level of agreement. RESULTS: The mean differences between the FIM motor and cognitive scales and their MDS-PAC translations were 2.4 (mean = 45) and 0.0 (mean = 28), respectively, with scale correlations of.85 and.84. Weighted kappas on individual items ranged from.32 to.64. There were substantial hospital specific differences in scoring. Payment cell classification using FIM data agreed with that using MDS-PAC data only 56% of the time. Twenty percent of the facilities experienced revenue shifts larger than 10%. CONCLUSION: Despite better item-level agreement than previously observed, poor payment cell agreement and substantial revenue shifts indicated that the MDS-PAC should not be substituted for the FIM trade mark instrument in the rehabilitation hospital PPS. PMID- 14734954 TI - Health-related quality of life among patients with breast cancer receiving zoledronic acid or pamidronate disodium for metastatic bone lesions. AB - BACKGROUND: Research on individual differences in health-related quality of life (HRQOL) can identify intervention targets and important covariates in analyses of treatment outcomes. OBJECTIVES: The objectives of this study were to describe HRQOL trajectories for women with metastatic breast cancer in a randomized trial of bisphosphonates and to identify characteristics associated with variations in HRQOL. RESEARCH DESIGN: We conducted a prospective quality-of-life study within a randomized, controlled trial. SUBJECTS: We studied women with metastatic breast cancer receiving zoledronic acid or pamidronate disodium to reduce the incidence of skeletal-related events (SREs). MAIN OUTCOME MEASURES: HRQOL was measured at fixed time points during the trial. Individual growth-curve modeling was used to describe longitudinal trajectories and to identify predictors of trajectories. RESULTS: For most domains of HRQOL, the mean trajectory reflected a mild increase, which leveled off later in the trial. Older age and full-time employment were associated with higher baseline HRQOL. Longer time from cancer diagnosis to randomization, lower Eastern Cooperative Oncology Group (ECOG) status (score of 2 ["inactive"]), and a history of SREs were associated with lower baseline HRQOL. Significant differences across geographic regions were observed for all domains. Active ECOG status (score of 0-1) at baseline was predictive of greater increases in all domains of HRQOL except Social/Family Well Being. Age, geographic region, and time from first bone metastases to randomization were associated with longitudinal changes in some domains. CONCLUSIONS: Women with metastatic breast cancer receiving bisphosphonates for prevention of SREs experienced an overall increase in HRQOL. Variations among women's experiences are explained partly by such characteristics as a history of SREs. PMID- 14734955 TI - Characteristics of occasional and frequent emergency department users: do insurance coverage and access to care matter? AB - OBJECTIVE: The objective of this study was to explore how insurance coverage, access to care, and other individual characteristics are related to the large differences in emergency department (ED) use among the general population. MATERIALS AND METHODS: We used the 1997 and 1999 National Survey of America's Families, a nationally representative sample. People were classified into 3 ED use levels based on the number of visits over the 12 months before the survey: non-ED users (zero visits), occasional users (1 or 2 visits), or frequent users (3 or more visits). We used a multinomial logit model to estimate the effect of insurance status and other factors on levels of ED use, and to compute the odds ratios of being occasional and frequent users as opposed to nonusers among various subpopulations. RESULTS: People in fair/poor health are 3.64 times more likely than others to be frequent ED users as compared with nonusers. The uninsured and the privately insured adults have the same risk of being frequent users, but publicly insured adults are 2.08 times more likely to be frequent users. Adults who made 3 or more visits to doctors are 5.29 times more likely to be frequent ED users than those who made no such visits. CONCLUSION: The uninsured do not use more ED visits than the insured population as is sometimes argued. Instead, the publicly insured are overrepresented among ED users. Frequent ED users do not appear to use the ED as a substitute for their primary care but, in fact, are a less healthy population who need and use more care overall. PMID- 14734956 TI - Discrete state analysis for interpretation of data from clinical trials. AB - OBJECTIVE: The objective of this study was to demonstrate a multivariate health state approach to analyzing complex disease data that allows projection of long term outcomes using clustering, Markov modeling, and preference weights. SUBJECTS: We studied patients hospitalized 30 to 364 days with refractory schizophrenia at 15 Veterans Affairs medical centers. STUDY DESIGN: We conducted a randomized clinical trial comparing clozapine, an atypical antipsychotic, and haloperidol, a conventional antipsychotic. METHODS: Health status instruments measuring disease-related symptoms and drug side effects were administered in face-to-face interviews at baseline, 6 weeks, and quarterly follow-up intervals for 1 year. Cost data were derived from Veterans Affairs records supplemented by interviews. K-means clustering was used to identify a small number of health states for each instrument. Markov modeling was used to estimate long-term outcomes. RESULTS: Multivariate models with 7 and 6 states, respectively, were required to describe patterns of psychiatric symptoms and side effects (movement disorders). Clozapine increased the proportion of clients in states characterized by mild psychiatric symptoms and decreased the proportion with severe positive symptoms but showed no long-term benefit for negative symptoms. Clozapine dramatically increased the proportion of patients with no movement side effects and decreased incidences of mild akathisia. Effects on extrapyramidal symptoms and tardive dyskinesia were far less pronounced and slower to develop. Markov modeling confirms the consistency of these findings. CONCLUSIONS: Analyzing complex disease data using multivariate health state models allows a richer understanding of trial effects and projection of long-term outcomes. Although clozapine generates substantially fewer side effects than haloperidol, its impact on psychiatric aspects of schizophrenia is less robust and primarily involves positive symptoms. PMID- 14734957 TI - Access to healthcare for adolescents. PMID- 14734958 TI - Should school-based services be routinely available to teen mothers? PMID- 14734959 TI - Should school-based services be routinely available to teen mothers? PMID- 14734960 TI - Living on the edge: the current phenomenon of self-mutilation in adolescents. AB - The purpose of this article is to share current knowledge about adolescent self mutilation (SM), and to discuss treatment approaches for affected teens and young adults. One in eight American teenagers experiences depression or anxiety. The growing phenomenon of SM is a particularly worrisome coping mechanism used by teens and young adults to deal with stress and sadness. The incidence of SM is increasing, and might be fueled by current trends in music and media that highlight violent and self-injurious behaviors. SM represents a "cry for help" by adolescents who are suffering. Self-mutilators who repeatedly cut themselves with razors, scissors, knives, or sharp glass are not attempting suicide, but are seeking to relieve extreme anxiety, tension, or pain. Medical literature regarding SM is limited, and evidence-based treatment approaches have not been documented; however, early discovery of SM and the development of a multifaceted treatment approach that incorporates the child, family, and trusted practitioners could be instrumental in managing SM. There is hope that antidepressant medications may also be beneficial. Research is needed to identify successful treatment approaches, and to discover the best ways to educate teens, families, educators, and community leaders about the prevalence of SM in our youth. PMID- 14734961 TI - Drug and substance use in adolescents. AB - This article provides an overview of adolescent drug and substance use, and includes prevalence and trends, commonly occurring comorbid conditions, clinical manifestations of drug and substance use, and evidence-based prevention and treatment principles. Risk and protective factors in five domains are also discussed in this article to provide guidance for assessment and care planning. A detailed table of the most prevalent drugs used by adolescents, including the drug's street names and the clinical manifestations of each drug's use, is offered to assist nurses in understanding their adolescent patients' language and to aid in teaching. Nurses are in varied and ideal positions to begin early screening (and to include families, peers, and other important influences in the adolescent's life), to provide continuity of care, and to advocate in the policy arena for development and funding of comprehensive and efficacious programs to help prevent or treat substance use in adolescents. PMID- 14734962 TI - New options for teen pregnancy prevention. AB - Teenage pregnancy rates remain unacceptably high in the United States, despite the availability of numerous contraceptive options. This article examines contraceptive methods currently available to adolescents and highlights newer products that may help meet some of the unique contraceptive needs of sexually active teens. The three newest agents (OrthoEvra trade mark, NuvaRing trade mark, and Seasonale) offer advantages such as higher contraceptive efficacy, increased privacy, ease of use, and low side effect profiles. A major advantage of each of these methods is that they eliminate the need for the teen to take daily action to prevent pregnancy. Nurses can be instrumental in helping teens to avoid unwanted pregnancy by assessing teens' contraceptive needs and educating them about contraceptive options. However, it is important to remember that because none of these new methods offer protection against sexually transmitted diseases, we should continue to educate adolescents about the need for condom use and provide them with instructions on using condoms correctly. PMID- 14734963 TI - Positive youth development: reducing the health risks of homeless youth. AB - This article outlines several preventive health strategies for reducing the health risks of homeless youth related to emotional distress, alcohol and other drug use/abuse, risky sex, and victimization, all of which are well documented as major health risks for homeless youth living on the street. These health risks interrupt normal adolescent development and are primary obstacles to exiting the street culture and lifestyle. Research indicates that risk exposures among adolescents can be moderated and/or buffered by a focus on individual strengths and environmental protective factors such as community support and mentoring. PMID- 14734964 TI - Common menstrual disorders in adolescence: nursing interventions. AB - Menstrual disorders such as amenorrhea, excessive uterine bleeding, dysmenorrhea, and premenstrual syndrome are common reasons for visits to healthcare providers by adolescent girls. Although menstrual irregularity can be normal during the first few years after menarche, other menstrual signs and symptoms may indicate a pathological condition that requires prompt attention and referral. This article discusses four common menstrual disorders seen in adolescent girls and focuses on specific nursing interventions aimed at eliciting an accurate menstrual history, providing confidentiality and communicating therapeutically, administering culturally sensitive care, and promoting independence and self-care. PMID- 14734965 TI - Use of home pregnancy tests among adolescent women. AB - Many adolescents use home pregnancy tests when they suspect pregnancy. However, because of developmental issues and greater variation in menstrual cycles, teens are at risk for obtaining false-negative test results. Moreover, with teens, the need for a pregnancy test often accompanies the need for comprehensive reproductive care including evaluation for sexually transmitted infections and contraception. This article reviews the mechanisms of home pregnancy testing including sensitivity, accuracy, and home testing procedures. Clinical strategies for improving the accuracy of home pregnancy tests for teen users are discussed. Lastly, policy implications to improve teens' access to more accurate pregnancy testing and to reproductive health services are suggested. PMID- 14734966 TI - Intergenerational family relations and sexual risk behavior in young women. AB - PURPOSE: To more fully understand the associations between family variables and sexual behavior of young women. STUDY DESIGN AND METHODS: Forty-two female clients of an urban youth agency (16 to 25 years of age) were interviewed about intergenerational stories and contact with extended families. Individuation was measured by self-report using the intergenerational individuation subscale of the Personal Authority in the Family System Questionnaire. Self-report questions were used to gain information about the young women's sexual risk behavior. Data were analyzed using Pearson correlation, independent t-tests, and multiple regression. Family stories were analyzed for themes of resilience or vulnerability. RESULTS: Young women who perceived their connection with previous generations in a resilient light tended to be more individuated and report less risky sexual behaviors. CLINICAL IMPLICATIONS: These results may give direction in the design of family interventions for reducing sexual risk taking and enhancing positive health behavior. Interventions would include promoting a healthy degree of attachment between adolescents and their families, exploration of family of origin issues, and referral for more intensive services when needed. PMID- 14734968 TI - Distractive flexion injuries of the subaxial cervical spine treated with anterior plate alone. AB - The clinical and radiographic effect of anterior plate fixation alone was evaluated in 36 consecutive patients with distractive flexion (DF) injuries in the lower cervical spine. Mean follow-up time was 15 months. The aim of the present study was to determine whether anterior plate fixation alone provides sufficient stability when treating DF injuries in the cervical spine. Solid union was seen in 6 of 6 patients with stage 1 injury and in 15 of 17 patients with stage 2 injury. In the patients with stage 3 injury, 7 of 13 of the anterior fixations failed. These failures occurred mainly among the patients with severe neurologic injuries. We believe these findings substantiate the use of anterior plate alone for DF injuries at stage 1 and 2 but disqualify anterior plate fixation alone for DF injuries at stage 3, with neurologic injury present. PMID- 14734969 TI - The external fixator: a tool for evaluation of complex low back pain problems. AB - BACKGROUND: The selection of patients with low back pain for fusion is especially difficult when previous surgical interventions failed and/or invasive tests (discograms, facet blocks) do not allow or do not clearly identify the painful motion segment. Test fixation with an external fixator may mimic a definite internal fixation, such as a fusion, and may help select patients for a more favorable result. The purpose of the study was to clarify if temporary back pain relief by external fixation is predictive for back pain relief after final internal fixation and fusion. METHODS: A retrospective study of 63 patients up to 6 years after evaluation of low back pain problems with an external fixator (index operation) was conducted. The data were collected from the charts based on a protocol that patients had to fill in during evaluation, and the actual state was checked during a clinical investigation at the outpatient clinic. The protocol monitored pain (Visual Analog Scale), pain medication, and work status. Based on the first two parameters, the effect of external fixation was defined as a general estimation as positive, doubtful, or negative. The same parameters were reassessed at follow-up control. RESULTS: In 38 patients, the immobilization test did improve the pain situation, in 11 it remained unchanged, and in 14 the pain got worse. Thirty-eight patients (not identical with the above) underwent definitive fusion afterward, whereas 25 were not operated on. Twenty-one of 30 patients with a positive effect undergoing surgery showed an improved situation after fusion, and 8 of 22 patients without further operation were improved at follow-up. Nine of 12 patients with a negative immobilization test did stay in a bad situation at follow-up. CONCLUSION: The external fixator as a tool for evaluation of patients with low back pain is an expensive measure with a considerable complication risk and only justified in selected patients when any other measure fails to assess and evaluate a patient's situation. If the test fixation reveals no benefit, the patients will remain in a bad situation whatever the therapeutic measure will be. Therefore the main value of the external fixator assessment is the selection of these patients that should not undergo surgery. A positive test fixation means a 72% chance for a satisfactory outcome at least two years after surgery, whereas without surgery the chance for some spontaneous improvement is 57% if the test immobilization did show some improvement. With respect of the "negative" selection of this group of patients (complex history, previous interventions) in our as well as in others series, the obtained results seem acceptable and the use of this invasive diagnostic measure in this group of patients seems justified. Statistical analysis did not show differences of significance as the numbers in the individual groups was too small. PMID- 14734970 TI - Commentary on the external fixator. PMID- 14734971 TI - Long-term outcomes of surgical treatment for tethered cord syndrome. AB - SUMMARY: Surgical outcomes for tethered spinal cord syndrome (TCS) associated with low-lying conus medullaris were evaluated. We investigated the long-term results of untethering the spinal cord and dural plasty in surgical patients with a wide age range. Improvement of bladder dysfunction and suppression of leg deformity progression were noted in two pediatric patients. However, severe urinary dysfunction generally remained postoperatively. Adult patients commonly showed low back or leg pain as clinical manifestations. In seven adult patients, urinary dysfunction also improved. Neurologic findings and urinary deficits showed a favorable improvement mostly in adult TCS in comparison with natal or juvenile onset of TCS. A short duration from onset to surgery and cranial movement of the conus medullaris as assessed by postoperative magnetic resonance imaging were factors indicating a favorable prognosis. PMID- 14734972 TI - New percutaneous access method for minimally invasive anterior lumbosacral surgery. AB - Preliminary cadaver, animal, and human studies were performed to determine the feasibility of axial anterior lumbosacral spine access using a percutaneous, presacral approach. Custom instruments were directed under fluoroscopic guidance along the midline of the anterior sacrum to the surface of the sacral promontory where an axial bore was created into the lower lumbar vertebral bodies and discs. Imaging and gross dissection were performed in cadavers and animals. The procedure was used for lumbosacral biopsy in human subjects guided by intraoperative imaging and clinical monitoring. All procedures were technically successful. This study demonstrates the feasibility of the axial access technique to the anterior lower lumbar spine. PMID- 14734973 TI - Thoracic pedicle screw insertion using a transpedicular drill guide: a preliminary study. AB - Insertion of thoracic pedicle screws can lead to major complications. This study reports the use of a transpedicular drill guide (TDG) for safe pedicle screw insertion in the thoracic spine. The conventional anatomic technique and the TDG were both used to drill pilot holes into the pedicles of four anatomic models of the thoracic spine. Ninety-nine percent of the 96 pilot holes drilled with the TDG were within 2 mm from the pedicle wall compared with 79% for the anatomic technique (P < 0.001). The TDG reduced the proportion and the extent of medial perforations. The TDG was easy to use and was superior to the conventional anatomic technique. It could be combined with fluoroscopy and pedicle palpation in certain clinical applications, especially for training surgeons, but only after confirming its accuracy in a cadaveric study. PMID- 14734974 TI - Correlation between backpack weight and way of carrying, sagittal and frontal spinal curvatures, athletic activity, and dorsal and low back pain in schoolchildren and adolescents. AB - This cross-sectional study was carried out to investigate any correlation between backpack carrying, spinal curvatures, and athletic activities on schoolchildren's dorsal (DP) and low back pain (LBP). Three thousand four hundred forty-one students aged from 9 to 15 years who carried backpacks to school were included in this study and asked for DP and LBP experiences in the school period while carrying the backpack. Nonradiating methods (surface back contour analysis) were used to indirectly measure frontal spinal curve (scoliosis) with the scoliometer and lateral curves (thoracic kyphosis and lumbar lordosis) with the kyphometer. All data analyses were undertaken regarding school year level, age, gender, sports participation, backpack weight, and way of carrying (one versus both shoulder) in relation to magnitude of scoliosis, thoracic kyphosis, lumbar lordosis, and DP and LBP while carrying the backpack. DP increased with increasing backpack weight (P < 0.05). The way (one versus both shoulder) of backpack carrying did not correlate either with DP or with LBP. Girls experienced much more LBP and DP than boys (P < 0.001). There was no difference in the prevalence of LBP and DP between adolescents and children. Students' age, height, and body weight as well as magnitude of kyphosis, lordosis, and scoliosis did not correlate with either LBP or DP. At the age of 11 years, girls and boys showed the highest prevalence for DP (72% and 38.5%, respectively), while at the age of 14 years, girls reported significantly (P < 0.05) more DP than boys. Girls showed the highest prevalence of LBP (71%) at the age of 11 years, while for the boys, it was at the age of 15 years (21%). Girls showed at the age of 11 years significantly more LBP (P < 0.05) than boys. Sports exposure seemed to increase LBP in girls (P < 0.001). The results of this study suggest a differential DP and LBP prevalence in schoolchildren and adolescents carrying backpacks with regard to gender and age. The peak in pain prevalence was immediately before puberty as well as immediately after its onset. Girls who participated in sports activities seem to experience more often DP and LBP than boys. Short children who carry backpacks as heavy as do tall children at the same age are more prone to LBP. PMID- 14734975 TI - Use of the ophthalmic phacoemulsification instrument for tumors of the spinal cord: report of seven cases. AB - In this retrospective cohort study, we examined the safety, efficacy, and benefits of utilizing the readily available ophthalmic phacoemulsification instrument for the surgery of seven intramedullary spinal cord tumors. Historically, the conventional neurosurgical ultrasonic aspirator was an adaptation of the original Cavitron Phaco-Emulsifier aspirator used in cataract surgery. The ophthalmic instrument has a 1.0-mm internal diameter operating tip. This is much smaller and provides improved visualization and surgical control when compared with the conventional neurosurgical instrument, which has a tip diameter of 3-4 mm. A satisfactory surgical and clinical outcome was achieved in all seven patients with safe and substantial subtotal excision of their tumors. We found the ophthalmic instrument provided excellent visualization and precision in operating on cases of intramedullary spinal cord tumor. PMID- 14734976 TI - Radiologic stability of titanium mesh cages for anterior spinal reconstruction following thoracolumbar corpectomy. AB - BACKGROUND: This work evaluated the radiologic stability of titanium mesh cages (TMCs) when used for single-level corpectomy reconstruction of thoracic and thoracolumbar spine. METHODS: Thirty-one patients underwent reconstruction for acute fractures (n = 15), posttraumatic deformity reconstruction (n = 10), neoplastic disorders (n = 4), and infection (n = 2). The cages were placed after corpectomy and excision of the adjacent intervertebral discs. Additional stabilization devices included anterior plates alone (n = 18), anterior double screw and rod constructs alone (n = 9), a single anterior rod system (n = 1), posterior stabilization alone (n = 6), and additional posterior stabilization (n = 2). RESULTS: Mean kyphosis correction was from 16 degrees to 5 degrees with 3 degrees of recurrence at 1-year follow-up (P < 0.0001 for both postoperative and final follow-up). In patients with greater initial kyphosis (>20 degrees ), mean correction was from 33 degrees to 10 degrees without recurrence (P = 0.004). Distance between adjacent vertebral bodies improved by 13 mm after cage placement, with a mean of 2mm of settling at final follow-up. There was one asymptomatic cage fracture without evidence of other problems. Two patients had construct failure after complex three-dimensional deformities were inadequately corrected and the cages had been placed in an angulated position. CONCLUSIONS: This report suggests that TMCs are a sound reconstruction alternative after thoracic and thoracolumbar corpectomy at a single level and may prevent complications associated with the harvest and use of large structural autografts for these reconstructions. Failure to correctly align the spine so the cage can be vertically placed is a contraindication to the use of TMCs. PMID- 14734977 TI - The effect of blocking a nutritional pathway to the intervertebral disc in the dog model. AB - BACKGROUND: The hypothesis that injecting bone cement adjacent to one or both endplates would bring about degeneration in the intervening disc was tested. METHODS: In 11 dogs, bone cement was injected just below the superior endplates of L1, L2, and L3 to block the nutritional supply through these endplates to the three intervertebral discs T13-L1, L1-L2, and L2-L3. In one other dog, both the superior and the inferior endplates of the same discs (T13-L1, L1-L2, and L2-L3) were blocked with bone cement. All 12 dogs were euthanized between 31 and 70 weeks after the surgery. The three experimental discs (T13-L1, L1-L2, and L2-L3) and two control discs (T12-T13 and L4-L5) were excised and assessed using enzyme linked immunosorbent assay (ELISA) and histology. RESULTS: Radiographs of the lumbar spine at the time of death did not show any signs of disc bulging, disc space narrowing, or peripheral osteophyte formation in any of the 12 dogs. The experimental discs as well as the control discs appeared normal in every dog. After the discs were bisected, they were carefully inspected for any visible signs of degeneration. The experimental discs showed no clear signs of disc degeneration and were not distinguishable from the control discs on a gross level. The numerical results from the ELISA showed that in the experimental discs as opposed to the control discs, there were significant increases in proteoglycan content in both the nucleus (P = 0.033) and annulus (P = 0.01) and clear histologic changes in some of the discs. CONCLUSION: The results show that injecting bone cement adjacent to one or both endplates for up to 70 weeks does not produce degeneration in any visible form in the intervening disc. There were no disc bulging, no apparent annular fissures, and no disc spacing narrowing. There were, however, increases in protoglycan content in both the nucleus and the annulus and clear histologic changes in some of the discs. PMID- 14734978 TI - Experimental disc degeneration due to endplate injury. AB - The aim of this study was to create an experimental model of disc degeneration that closely mimicked human disc degeneration. In six domestic pigs, an L4 cranial endplate perforation into the nucleus pulposus was made. Three months postoperatively, compressive testing was performed on the L2-L4 motion segments, and intradiscal pressure was measured in the intervening discs. Histochemical and morphologic examinations were made on the excised degenerated and adjacent discs. A significant reduction in water content was observed in the outer anterior annulus of the degenerated disc. In the nucleus, the proteoglycan content was significantly reduced, as well as the cellularity, although not significantly. The nucleus lost its gel-like structure and was discolored, and there was delamination of annular layers. Intradiscal pressure in the nucleus was significantly lower in the degenerated disc. In conclusion, experimental degeneration of the intervertebral disc induced by endplate penetration resembled human disc degeneration, as exemplified by biochemical and structural changes. PMID- 14734979 TI - An interspinous process distractor (X STOP) for lumbar spinal stenosis in elderly patients: preliminary experiences in 10 consecutive cases. AB - BACKGROUND: Lumbar spinal stenosis (LSS) is often a position-dependent condition that is aggravated in extension and relieved in flexion. METHODS: Ten consecutive elderly patients with LSS were assessed postoperatively by magnetic resonance imaging and the Swiss Spinal Stenosis Questionnaire. Cross-sectional areas of the dural sac and intervertebral foramina at the stenotic level were measured postoperatively and compared with the preoperative values. RESULTS: Postoperatively the cross-sectional area of the dural sac increased 16.6 mm2 or 22.3% and intervertebral foramina increased 22 mm2 or 36.5%. The intervertebral angle and the posterior disc height changed significantly. Seventy percent of the patients were satisfied with the surgical outcome. CONCLUSION: This new surgical method is effective in elderly LSS patients. PMID- 14734982 TI - Test your knowledge: antineoplastic therapy. AB - The CRNI Certification Examination consists of 200 multiple-choice questions covering the nine core content areas of infusion nursing: Technology and Clinical Applications, Fluid and Electrolyte Balance, Pharmacology, Infection Control, Pediatrics, Transfusion Therapy, Antineoplastic Therapy, Parenteral Nutrition, and Quality Assurance. The review questions provided below are modeled on the CRNI exam and are intended to help exam candidates test their knowledge of infusion therapy practice. This special section is a regular addition to the Journal of Infusion Nursing, with each edition focusing on a single content area. PMID- 14734983 TI - Complications associated with drug and nutrient interactions. AB - Patients receiving parenteral nutrition (PN) usually have conditions that require intravenous (IV) medications. Antibiotics, and cardiovascular, gastrointestinal and electrolyte agents are commonly prescribed for these patients. When the gastrointestinal tract is compromised, parenteral administration of these agents is often necessary. Because of the complex nature of PN and IV medications, a potential exists for serious or life-threatening drug and nutrient interactions to occur if procedures are not in place to foster their safe administration. This article will describe the physico-chemical characteristics of PN and concepts used in dealing with issues of drug and nutrient interactions that occur in patients requiring PN. PMID- 14734984 TI - Perception of risk factors for infusion phlebitis among Swedish nurses: a questionnaire study. AB - This questionnaire study was set up to assess the perceptions of risk factors for infusion phlebitis among Swedish nurses, as their concepts of these factors may influence the incidence. A majority of the nurses believed that insertion of a peripheral venous catheter in the forearm and catheter rotation within 48 hours was protective. These measures are not supported in recent studies and guidelines. Surveillance of the educational level of staff, who insert peripheral venous catheters, is an important tool for reducing the incidence of infusion phlebitis. PMID- 14734985 TI - The Safe Medical Device act and its impact on clinical practice. AB - Hospital patients often require vascular access devices to deliver infusion therapies. These can include peripheral intravenous catheters (PIVs), peripherally inserted central venous catheters (PICCs), triple lumens, ports, and Tesio, Swann-Ganz, and Hickman catheters. Although these products are used in large numbers without significant sequelae, when a device fails, the impact is realized throughout the industry. The infusion clinicians within our institution followed a systematic approach to dealing with the failure of devices that occurred over a 3-month period. This article describes the response process when a device fails and the methods of achieving positive patient outcomes. PMID- 14734987 TI - Nutritional assessment and support of kidney transplant recipients. AB - Kidney transplant has become a viable option for patients with end-stage renal disease (ESRD). The number of kidney transplants has steadily increased during the past 50 years. Advances in surgical technique and immunosuppressive drugs have led to significant improvements in survival rates. Many chronic diseases that lead to ESRD negatively affect nutritional status. To minimize nutritional depletion and optimize nutritional status, a complete and thorough evaluation by a registered dietitian should be performed. The posttransplant nutritional goal is to provide adequate nutrition to promote wound healing and anabolism, to prevent infection, and to minimize side effects of medications. Providing adequate nutrition and reducing the long-term side effects are essential for graft survival in kidney transplant recipients PMID- 14734986 TI - Evaluation of a no-dressing intervention for tunneled central venous catheter exit sites. AB - This study tested whether central venous catheter (CVC)-related sepsis could be reduced by removing a hypothesized reservoir for pathogens, the CVC exit site dressing. Seventy-eight individuals with cancer, stratified for gender (37 men and 41 women) and transplant status, with newly inserted CVCs were recruited and randomly assigned to receive either a gauze dressing or no dressing, once their catheter insertion site had healed (3 weeks). Because there was no difference in CVC-related septic episodes based on gender or transplant status, the stratification was not maintained for remaining analyses. Although there was no significant difference in CVC-related septic episodes (P =.28) or rehospitalization rates (P =.41) because of CVC-related sepsis between the dressing and no-dressing group, individuals in the dressing group developed CVC related sepsis sooner (P =.02) than did individuals in the no-dressing group. PMID- 14734988 TI - Individually dissecting each health policy issue negatively effects the entire healthcare system. PMID- 14734989 TI - OASIS diagnosis reporting: case examples. PMID- 14734991 TI - Why won't this wound heal, and what should I do about it? PMID- 14734992 TI - Patient outcomes terminology: what does it all mean? Why do I need to know? PMID- 14734993 TI - Using the nursing process to accurately plan the Medicare 60-day episode of care: you really do know how to do this! PMID- 14734994 TI - Researching end-of-life care: challenges, strategies, and opportunities. PMID- 14734995 TI - A review of cardiac anatomy and physiology. PMID- 14734996 TI - Low-dose aspirin interactions. PMID- 14734997 TI - What's new in the 2004 Joint Commission home care and hospice standards? Part 1. PMID- 14734998 TI - Choosing and using a digital camera in home care. PMID- 14734999 TI - A nurse's fantasy. PMID- 14735000 TI - [Non traumatic spinal epidural hematomas. Report of twenty cases. Review of the literature and outcome aspects]. AB - BACKGROUND AND PURPOSE: We report a personal series of 20 non traumatic spinal epidural hematomas and study outcome aspects with a review of data in the literature. METHOD: Clinical presentation of non-traumatic spinal epidural hematomas observed between January 1980 and December 1998 was acute in 17 cases (85%) and chronic in 3 (15%). Symptoms were spinal and/or radicular pain, sensorimotor and sphincter dysfunction. Radiological evaluation consisted in myelography (n=6), myelography-CT scan (n=5), CT scan (n=1) and MRI (n=9). Patients underwent surgery in 15 cases, between 8 hours and 2 months after the first symptoms. All our patients were clinically reevaluated between 2 and 4 months after either surgery or admission for cases of spontaneous resolution. RESULTS: Good results (complete neurological resolution or moderate sequelae) were observed in 14 patients (70%). A partial recovery with major persistent neurological impairment was observed in 1 patient (5%), an initial persistent neurological impairment in 1 (5%). Three patients (15%) died and 1 (5%) was lost to follow-up. Complete spontaneous resolution were observed in four patients. CONCLUSION: Postsurgical outcome is mainly related to the preoperative neurological impairment, the duration of spinal cord compression and the time interval between the onset of symptoms and maximal deficit. A prompt laminectomy is necessary except in the cases where a spontaneous resolution can be expected from the early neurological course. PMID- 14735001 TI - [Anterior cervical discectomy with or without bone graft placement in the treatment of cervical radiculopathy. Long-term results]. AB - BACKGROUND AND PURPOSE: Anterior approach for cervical radiculopathy is a frequently performed operation in neurosurgery. The goal of this study is to evaluate the short and long-term results of anterior cervical discectomy with and without fusion. METHODS: Between 1984 and 1999, we operated on 101 patients presenting with cervical radiculopathy by an anterior approach. The operation consisted of one-level discectomy in 74 cases, two-level discectomy in 25 cases and three-level discectomy in 2 cases. Eighty-four out of 130 levels operated on were fused. Evaluation was done following a consultation with dynamic cervical X rays and by telephone using a detailed questionnaire. Follow-up was obtained in 91 cases with a mean of 54 months in fused patients, and a mean of 45 months in the non-fused patients. RESULTS: Post-operative results were good in 95% in the 2 groups of patients. We encountered 8 complications, all in patients operated with bone graft placement. Five patients were reoperated on at an adjacent level, 4 being fused previously. CONCLUSIONS: The 2 techniques are comparable in term of goods results at short and long-term follow-up. Anterior discectomy without fusion is associated with less complications, less post-operative pain, and less operative time cost. PMID- 14735002 TI - [Update on cellular and molecular biology techniques]. AB - Cellular and molecular biology techniques have invaded all fields of medicine and neurosurgery is not an exception as can be seen at congresses and in our journals. The purpose of the present review is to provide an update on current techniques. PMID- 14735003 TI - [Update on medical management of severe head trauma]. AB - Significant progress in prognosis after brain injury has been achieved over the last 20 years. Knowledge of post-traumatic brain hypersensitivity to ischemic events is critical for management. Therefore, all recommended emergency treatments (intubation, oxygenation, prevention of hypotension) focus on situations where oxygen delivery to the brain is compromised (peripheral hypoxia or hypotension but also compressible cerebral hematoma). Analysis of European prehospital medical care showed success in peripheral oxygenation but no real benefit regarding blood pressure. Guidelines for osmotherapy in patients with pupil abnormalities are not followed despite recent studies emphasizing better prognosis after acute perfusion of high-dose mannitol followed by rapid surgical treatment. It is well known that a short delay between trauma and surgery improves prognosis. After controlling peripheral hemodynamics and hemostasis, multimodal monitoring (intracranial pressure, transcranial Doppler, SvjO(2)) is necessary to achieve cerebral hemodynamic equilibrium. Management during the first hours after trauma is important for outcome in patients with traumatic brain injury. A well-organized medical referral system with close collaboration between specialists will be able to control this socially accepted silent epidemic. PMID- 14735004 TI - [Update on neurotraumatology]. PMID- 14735005 TI - [Tophaceous gout of the cervical spine, causing cord compression. Case report and review of the literature]. AB - We present the case of a 74-year-old patient with long-standing gout who developed C4-C5 tophaceous gout causing cord compression. The patient had undergone 4 years earlier for a disco-osteophytic cord compression (anterior C4 C5 and C5-C6 discectomy). When admitted, the patient presented quadriparesia which had developed during the previous week in association with acute tophaceous gout on the knees, and the metarsophalangeal articulations, in the context of a bronchial infection. The diagnosis of spondylodiscitis was initially retained because of the clinical features of severe infection and the radiological data (C4-C5 cord compression, with anterior epidural lesions in MRI). The intervention allowed decompression and pathological diagnosis of tophaceous gout. Spinal gout is well-known, and very rarely responsible for cord compression: only 15 cases of cervical gout have been described in the literature. Radiological findings are not specific, and treatment is surgical in the event of medullar compression despite medical treatment. PMID- 14735006 TI - Tanycytic ependymoma of the spinal cord. Case report and review of the literature. AB - OBJECTIVE AND IMPORTANCE: We report a rare case of the tanycytic variant of intramedullary ependymoma. Tanycytes are the common progenitor cells of both ependymal cells and astrocytes. These particular elongate unipolar and bipolar ependymal cells extend from the ventricular lumen to the surface of the nervous system. It is extremely important, in terms of both management and prognosis, to distinguish intramedullary tanycytic ependymomas from intramedullary astrocytomas although a correct histological diagnosis may be difficult since tanycytes resemble astrocytes. CASE REPORT: A 39-year-old woman underwent surgical treatment for a cervical intramedullary tumor in our department. Although pathological examination of frozen sections was suggestive of low-grade astrocytoma, the definitive histological diagnosis was "tanycytic" ependymoma, a tumor characterized by poor cellularity, elongated elements mixed with fibrillary components, rare pseudo-rosettes and mixed astro-ependymal aspects. Since a complete resection was performed at surgery, no further treatment was proposed. After a follow-up period of two years the patient is free from recurrence. CONCLUSION: Tanycytic ependymomas should be managed in the same way as "ordinary" ependymomas, since there is no current evidence suggesting that these morphologically distinct tumors differ in terms of biological behavior. Increased awareness of this transitional form of intramedullary ependymoma among neurosurgeons and pathologists may avoid incorrect surgical approaches and postoperative treatment. PMID- 14735007 TI - [Intracranial aneurysm revealed by panhypopituitarism. Considerations on therapeutic management based on a case report and review of the literature]. AB - We report a case of an unruptured intracranial aneurysm in paraclinoidal location which was a unmasked by severe panhypopituitarism. An ipsilateral internal carotid artery (ICA) clamp test was performed in the pretreatment stage. The test was well tolerated and the aneurysm could be embolized with GDC (Guglielmi detachable coils) without necessitating an occlusion of the ICA. The postoperative course was uneventful and the patient was discharged after 48 hours. At one year, cerebral angiograms confirmed complete obliteration of the aneurysm. Therapeutic management of this rare entity is discussed along with a careful and exhaustive review of the literature. PMID- 14735008 TI - [Therapeutic management of lumbar synovial cysts. Comments on the article by Ph. Metellus et al]. PMID- 14735009 TI - Ground-glass opacity: interpretation of high resolution CT findings. AB - Ground-glass opacity (GGO) is a common finding on high resolution CT, characterised by areas of hazy increased attenuation of the lung with preservation of bronchial and vascular margins; it is not to be confused with consolidation, in which bronchovascular structures are obscured. It correlates with several pathogenic processes, such as like partial filling of air spaces, inflammatory or fibrotic interstitial thickening, increased capillary blood volume. Infiltrative GGO can representing either interstitial or alveolar processes. GGO is a nonspecific finding; however, the correlation with any of the associated CT findings (nodular lesions, consolidation, septal thickening, fibrosis, vessels or airway calibre alterations, air trapping), and clinical data is helpful in narrowing the range of diagnostic possibilities, or even in suggesting a specific diagnosis. GGO can indicate a potentially treatable disease, help guide the type and location of biopsy and evaluate the effectiveness of therapy. This review discusses the types of lung disease associated with GGO, and the differential diagnosis between GGO caused by infiltrative processes and the mosaic patterns of lung attenuation caused by primary vascular diseases or airway abnormalities. This distinction can be made by evaluating the vessel calibre and air trapping on expiratory scans. PMID- 14735010 TI - Thoracic manifestations of the less common collagen diseases. A pictorial essay. AB - Systemic autoimmune diseases include different forms of vasculitides and collagen diseases. Among collagen diseases, the rarer entities include: Sjogren syndrome, dermatopolymyositis, ankylosing spondylitis, relapsing polychondritis and mixed connective tissue disorders. The diagnosis of these entities requires an integrated multidisciplinary approach. The radiological findings of collagen diseases are well known; however, the introduction of HRCT studies provides additional information and enable an early diagnosis. The more common thoracic manifestations of collagen diseases include interstitial pneumonia (usual, nonspecific and lymphocytic), bronchiolitis obliterans organizing pneumonia (BOOP), airway diseases (bronchiectasis, obliterative and constrictive bronchiolitis, follicular bronchiolitis), pleural abnormalities, diaphragmatic dysfunction, apical fibrosis. The aim of this pictorial essay is to present the main radiological and HRCT patterns related to the less common collagen diseases. PMID- 14735011 TI - Digital mammography: physical principles and future applications. AB - Mammography is currently considered the best tool for the detection of breast cancer, pathology with a rate of incidence in constant increase. To produce the radiological picture a screen film combination is conventionally used. One of the inherent limitations of screen- film combination is the fact that the detection, display and storage processes are one and the same, making it impossible to separately optimize each stage. These limitations can be overcome with digital systems. In this work we evaluate the main characteristics of digital detectors available on the market and we compare the performance of digital and conventional systems. Digital mammography, due to the possibility to process images, offers many potential advantages, among these the possibility to introduce the dual-energy technique which employs the composition of two digital images obtained with two different energies to enhance the inherent contrast of pathologies by removing the uniform background. This technique was previously tested by using synchrotron monochromatic beam and a digital detector, and then the Senographe 2000D full-field digital system manufactured by GE Medical Systems. In this work we present preliminary results and the future applications of this technique. PMID- 14735012 TI - Preoperative staging of gastric carcinoma with multidetector spiral CT. AB - PURPOSE: To assess the accuracy of Multidetector computed tomography (MDCT) in the preoperative staging of gastric cancer. MATERIALS AND METHODS: Between March 2002 and October 2002, 27 patents with histologically proven gastric adenocarcinoma underwent MDCT. Unenhanced and contrast-enhanced CT scans were obtained after the oral administration of 400-600 ml of water for gastric wall distension. Biphasic enhanced scans were performed after the automatic injection of 2ml/kg of contrast agent at a flow rate of 3.5 ml/sec with a scan delay of 35 and 70 sec. The images were evaluated for: lesion morphology, degree of wall infiltration, presence of locoregional lymphadenopathies and distant metastases. Based on the findings, a TCMD staging system was established according to the criteria reported in the literature. All the patients underwent surgery, and the preoperative MDCT staging was evaluated against the pathology findings. RESULTS: MDCT staging was correct in 17/27 patients (62.9%). The T parameter was correctly assessed in 24/27 cases (88.9%), whereas it was understaged in 1 case (3.7%) (T1 stage at CT vs T2 at surgery) and overstaged in 2 cases (7.4%) (T3 vs T2). The N parameter was correctly evaluated in 19/27 patients (70.4%), understaged in 6/27 (22.2%) and overstaged in 2/27 (7.4%). CONCLUSIONS: MDCT may be proposed for the staging of gastric carcinomas and, although accuracy in N staging remains low in comparison to single-detector spiral CT, it provides a larger amount of diagnostic information. PMID- 14735013 TI - Malignant fibrous histiocytoma of the stomach. AB - MFH is a kind of sarcoma, normally located in the soft tissues of the extremity and retroperitoneal space. The involvment of the gastroenteric tube and particularly of the stomach are rather rare. Our case shows some differences from the others described in literature: the sight of mass, the age of the patient, the extension at the moment of the diagnosis. PMID- 14735014 TI - Air-fluid level in the ureter? It's possible with emphysematous pyelonephritis. PMID- 14735015 TI - MR Arthrography: a proposal for solution optimization with lidocaine. An in vitro experience. AB - PURPOSE: Pain on capsule distension in painful joints may affect feasibility of the MR Arthrography. We tried to overcome this limitation by adding a local anesthetic (lidocaine) to the paramagnetic contrast agent solution. We aimed at: a) investigating which contrast agent dilution provides the best signal-to-noise ratio in the SE T1 sequences; b) evaluating the effects of lidocaine on the signal intensity and on the viscosity of the solutions; assessing the viscosity of solutions containing iodinated contrast agent. MATERIALS AND METHODS: The paramagnetic contrast agent was diluted with saline and lidocaine at various concentrations. Signal intensity was measured with a 1.5 Tesla superconductive MR unit with a dedicated head coil; we used T1-weighted spin-echo sequence. The viscosity coefficient of the solutions was analyzed and compared with that of solutions containing iodinated contrast agents (but not lidocaine). RESULTS: Signal intensity is also unaffected by variations in the concentration of lidocaine, which does not interfere with the biphasic behavior of Gadolinium. Viscosity is scarcely affected by changes in lidocaine concentration when the paramagnetic contrast agent concentration is not changed. CONCLUSIONS: The optimal signal-to-noise ratio in T1-weighted sequences is provided by 0.4%, contrast agent dilution but contrast agent-saline solutions, with(out) lidocaine, cannot be considered steady and signal intensity values change over time. The addition of lidocaine does not significantly influence the signal-to-noise ratio and the viscosity of the solutions. The low viscosity of the paramagnetic contrast agent appears to favor quicker spread of the solution, even in tiny defects; thanks to its anesthetic effect, lidocaine could facilitates execution of the examination in painful joints without affecting the diagnostic result. PMID- 14735016 TI - Actual role of unenhanced magnetic resonance angiography (mra tof 3d) in the study of stenosis and occlusion of extracranial carotid artery. AB - PURPOSE: To evaluate the diagnostic accuracy, sensitivity and specificity of MRA (3D TOF) in comparison to contrast-enhanced MRA using DSA as the gold standard in the study of steno-occlusive disease of the internal carotid artery, in order to determine if this technique still has a diagnostic value. MATERIALS AND METHODS: Three radiologists blindly and independently evaluated either the unenhanced MRA, CE MRA, or DSA images of 23 consecutive patients, assessing the stenosis based on a NASCET five-class classification. RESULTS: The sensitivity, specificity, and diagnostic accuracy of CE MRA and unenhanced MRA were 100% and 100%, 100% and 95.5% and 100% and 93.2%, respectively. These differences were not statistically significant. CONCLUSIONS: Unenhanced MRA should be still considered a valid alternative for studying the extracranial section of the internal carotid arteries especially in patients who refuse or whose laboratory results exclude the use of contrast medium or in patients in whom continuous monitoring of a clinical condition is required but a repeated and/or frequent use of contrast medium is not possible. PMID- 14735017 TI - Multislice CT angiography in the assessment of peripheral aneurysms. AB - PURPOSE: Peripheral aneurysms are usually located in the femoro-popliteal district; these lesions may present some complications such as distal embolisation and thrombosis; rupture is quite rare, but may be life-threatening. The aim of this study is to test the accuracy of CT multislice angiography in the assessment of this pathology and to compare it with DSA, until now considered the gold standard in vascular imaging. MATERIALS AND METHODS: From January 2001 to May 2002 we studied 10 patients (8 males and 2 females) aged between 48 and 74 years with 14 lesions. All patients were affected by femoro-popliteal aneurysms. Eight patients underwent US as a first examination, 2 patients directly underwent DSA because of acute ischaemic symptoms; then, a CT angiography was performed. The first eight patients underwent CT and DSA after US. Hence, all patients underwent both DSA and CT. All CT examinations were performed with a multislice spiral CT scanner using the following parameters: 2 mm slice thickness, pitch 6, 2mm slice thickness, 1mm reconstruction interval, 120 mAs, 120 kVp. A nonionic contrast medium was infused intravenously at a biphasic rate. Angiography was performed by humeral artery catheterisation with a 4F device, the distal tip of the catheter was placed in the infra-renal abdominal aorta. RESULTS: In 7/10 patients the diagnosis was correctly formulated after DSA; in two patients, only the obstruction of the popliteal artery was detected, but not the dilatation and the thrombus. In one patient the parietal annular thrombus simulated a normal artery. CT was diagnostic in all cases and all the complications were detected as well. DISCUSSION AND CONCLUSIONS: Multislice spiral CT angiography allows depiction of aneurysms, and of the precise site, dimensions, quantity and quality of parietal thrombus; multislice CT is to be considered, in our opinion, essential in the assessment of diagnosis and in the planning of therapy. Moreover, CT provides exact measurements of the vessels before and after the aneurysm, which is essential to plan stent-graft implantation. A very interesting feature comes from the execution of a complete 3D angiography of the entire lower extremity circulation with high spatial resolution. DSA cannot provide direct information about the wall of the vessel and about the thrombus; therefore, in some cases it is unable to provide the correct diagnosis. Today DSA remains the best examination to evaluate the quality of run-off in lower extremity, especially under the knee. PMID- 14735018 TI - Endovascular treatment of intracranial arterio-venous malformations with Onyx embolization: preliminary experience. AB - PURPOSE: Intracranial arterio-venous malformations (AVM) often represent a complex clinical problem as regards indications to treatment, the choice of treatment and the technical difficulties related to treatment. In the last twenty years, endovascular treatment of intracranial arterio-venous malformations was primarily based on an acrylic glue (Hystoacril) whose endovascular use is not free from risks. A new product for endovascular embolization, named ONYX, has been recently been made commercially available. It is a bio-compatible liquid polymer that precipitates and solidifies in contact with blood, thus forming a soft and spongy embolus. The aim of this study was to evaluate our preliminary results in the endovascular embolization of intracranial AVM with Onyx. MATERIALS AND METHODS: Ten patients were treated (7 men, 3 women; mean age: 29 years, range: 12-48 years) for a total of 37 embolizations, 22 with Onyx and 15 with acrylic resin. RESULTS: Nidus occlusion was complete in 2 cases, >90% in 2 cases, >50 and <90% in 3 cases and <50% in the remaining 3 cases. The following complications were recorded: two transient and one mild permanent neurological deficits, two clinically silent cases of moderate subarachnoid haemorrhage, four catheters glued to the injection site, three cases of treatment discontinuation due to continuous and massive reflux of Onyx into the afferent artery peduncle. CONCLUSIONS: This early experience showed that while Onyx has good embolization potential it also presents some disadvantages, which need to be overcome before this product can be considered easy and safe to use on a large scale. PMID- 14735020 TI - Prognostic value of hyperattenuating middle cerebral artery sign at CT in cerebral infarction. AB - PURPOSE: To assess the prognostic value of hyperattenuating middle cerebral artery sign at CT in acute cerebral infarction. MATERIALS AND METHODS: Ninety-two patients with acute cerebral infarction in the sylvian area were retrospectively reviewed. All patients underwent unenhanced CT 12-24 hours after the onset of symptoms and follow-up CT within 48-72 hours. Initial CT scans were evaluated by consensus by three radiologists to confirm or exclude the presence of hyperattenuating middle cerebral artery sign. History, cardiovascular risk factors and neurological impairment at discharge (mean 25 days) were recorded for each patient. The degree of disability was graded 0 (no disability) to 6 (death). Patients were divided into two groups: patients without (group A) and patients with (group B) hyperattenuating middle cerebral artery sign. A logistic regression analysis was performed to compare the two groups. Results were correlated with cardiovascular risk factors. Kaplan-Meier survival curves were calculated for each group. RESULTS: Hyperattenuating middle cerebral artery sign was present in 18 patients (19.6%) (group B). The percentage of neurological deficits was significantly higher in group B than in group A (p<0.05). Sixteen (88.9%) of the 18 patients in group B had a poor prognostic index (score 3 - 6) with a significantly higher percent difference (41.6%, p<0.05) than patients of group A. Within 10 days of admission, 3 patients (16.7%) died in group A and 6 (8.1%) in group B. However, no significant differences were observed in the Kaplan-Meier survival curves. No correlation with cardiovascular risk factors was observed. CONCLUSIONS: Besides having an important diagnostic value, hyperattenuating middle cerebral artery sign is a reliable predictor of prognosis in terms of disability rather than mortality, as it reflect the larger extension of infarction. PMID- 14735019 TI - Diffusion-weighted MR Imaging: clinical applications in neuroradiology. AB - Among functional magnetic resonance imaging techniques, diffusion-weighted imaging (DWI) plays an important role in the assessment of a wide variety of brain diseases. DWI provides image contrast that depends on the molecular motion of water and it can be easily added to a standard cranial MR examination, with limited increase in time (imaging time ranges from a few seconds to 2 minutes). DWI is particularly sensitive in the detection of acute ischaemic stroke and in monitoring its evolution (also in the light of new therapeutic strategies for early treatment). Diffusion-weighted MR imaging also provides adjunctive information in the differential diagnosis of other brain diseases including neoplasms, intracranial infections, traumatic brain injury and demyelination some of which may manifest with sudden neurological deficits mimicking acute ischaemic stroke. PMID- 14735021 TI - Subarachnoid anaesthesia in caesarean delivery: effects on alertness. AB - AIM: Subjects in spinal anaesthesia have been reported to show a decrease in the level of alertness, even when they have not received any sedative drugs. THE AIM: of this study is to verify, in caesarean delivery if the bupivacaine subarachnoid anaesthesia, with or without intrathecal fentanyl dose, produces a sedative effect, to define the entity and to identify the mechanism that most likely causes it. METHODS: The clinical investigation is divided into Part I (non randomized trial) and Part II (randomized trial) and is set in the University hospital's delivery-unit. Part I: 45 pregnant women were recruited and enrolled in 3 groups: women having natural delivery (n=15), women receiving caesarean delivery with subarachnoid anaesthesia (n=15) and with general anaesthesia (n=15). Self-rating depression scale (SDS), self-rating anxiety scale (SAS), state anxiety inventory (SAI) and trait anxiety inventory (TAI) psychometric tests were patient-completed for pre-delivery anxiety evaluation. Part II: 23 pregnant women undergoing caesarean delivery were single-blind randomized to receive subarachnoid anaesthesia with single 12.75 mg 0.5% hyperbaric bupivacaine (n=10) or with 12.75 microg 0.5% hyperbaric bupivacaine and 15 mg fentanyl (n=13). Bispectral index (BIS), observer's assessment of alertness/sedation (OAA/S) scale, self-sedation visual analogic scale (VAS) and mean arterial pressure (MAP) were perioperatively monitored. RESULTS: Part I: it was found, using SAI, a higher level of state anxiety in the pregnant women undergoing caesarean delivery than those having natural delivery (p<0.05). Part II: pregnant women receiving subarachnoid anaesthesia for caesarean delivery had a decrease of the level of alertness from 10 to 70 min after the execution of lumbar puncture (p<0.05), with a sedative peak from 35 to 45 min (p<0.01), as measured by OAA/S scale and self-sedation VAS but not by BIS. Women who received bupivacaine fentanyl spinal anaesthesia had a more consistent sedative effect from 35 to 70 min (p<0.05). CONCLUSIONS: Pregnant women undergoing caesarean delivery had a more elevated level of state anxiety, assessed by SAI, as seen in Part I. Subarachnoid anaesthesia in caesarean delivery is associated with a reduction of alertness level. A double mechanism might cause a clinically valuable sedative effect, observed by OAA/S scale and self-sedation VAS: decrease of the afferent spinal inputs and an anxiolytic psychophysiologic effect, induced by good outcome of the birth and mother-baby contact. Intrathecal bupivacaine-fentanyl dose produces a larger alertness decrease than single bupivacaine, because the anaesthetic block density increases. BIS was not a sensitive measure to detect the light sedation occurring in Part II of this study. PMID- 14735022 TI - Ciaglia percutaneous dilational tracheostomy. Early and late complications and follow-up. AB - AIM: Evaluation of the safety of percutaneous dilational tracheostomy (PDT) for perioperative, early and late complications. METHODS: DESIGN: we prospectively collected complications in patients who underwent PDT for mechanical ventilation; patients were interviewed 8 months after discharge, symptomatic cases underwent ENT control. SETTING: 10 bed general ICU in a 650 -bed general hospital treating 450 patients per year. PARTICIPANTS AND INTERVENTION: 181 patients admitted between July 1998 and June 2000 who underwent PDT for mechanical ventilation. Prospe-ctive collection of data on patients and procedures and screening by a phone interview for symptoms possibly related to the tracheostomy. Symptomatic patients were referred to the ENT specialist. RESULTA: We found 17 perioperative minor complications and 10 minor during hospital stay complications. We traced 83 patients, alive 8 months after discharge. Sixty-one patients (73.5%) were symptom free. Four (4.8) complained of minimal dysphonia. Eighteen patients (21.7%) complained of symptoms deserving ENT control. Eleven patients came to the ENT control that was positive in 5 cases. In 2 patients swallowing uncoordination was found, in 1 arytenoid movement uncoordination. In 1 case (1.2%) a 25% tracheal stenosis was found. The stenosis was asymptomatic. One patient (1.2%) had a severe tracheal stenosis and had a Montgomery tracheal stent in place. CONCLUSIONS: In our experience Ciaglia PDT had an overall low rate of complications (21.8%). No patient had severe early complication. We found only 1 (1.2%) severe late complication. In selected patients, Ciaglia PDT with endoscopic control guarantees a high safety standard. PMID- 14735023 TI - Non-invasive mechanical ventilation in patients with acute cardiogenic pulmonary edema. AB - AIM: To evaluate the use of noninvasive mechanical ventilation (NIMV) in patients with acute cardiogenic pulmonary edema. METHODS: DESIGN: prospective study. SETTING: Emergency Department at a University hospital. PATIENTS: 84 patients with acute respiratory distress due to pulmonary edema. Interven-tions: NIMV, using a pressure support mode and positive end-expiratory pressure (PEEP). A "weaning test" to evaluate clinical stability. MEASUREMENTS: heart rate, arterial blood pressure, respiratory rate, arterial blood gases, electrocardiogram and incidence of myocardial infarction before and after NIMV. Mortality and duration of hospital stay were also considered. RESULTS: A total of 84 patients received NIMV with 14+/-3.6 cm H2O pressure support over PEEP of 8.3+/-2.1 cm H2O and FiO2 1. At the end of the study period, 16 patients (19%) were considered "non responders" and required invasive ventilation; 62 patients (74%) were considered "responders" and subsequently transferred to the medical ward. The hospital mortality was 14% and 25% in the "responder" and "non responder" groups, respectively; the length of stay was 15.7+/-10.1 days in the "responder" group vs 16+/-10.6 days in the "non responder" group. We never found new episodes of myocardial infarction related to NIMV. The only significant difference between "responder" and "non responder" patients was arterial blood pressure. CONCLUSIONS: We hypothesize that "non responder" patients, characterized by blood pressure values lower than "responders", are less "cardiocompetent" and thus unable to cope with the increased work of breathing. NIMV avoided Intensive Care Unit admission for 74% of the observed patients. PMID- 14735024 TI - High frequency percussive ventilation (HFPV). Principles and technique. AB - In recent years, the usefulness of high frequency ventilation (HFV) has been clinically reassessed as an alternative to conventional mechanical ventilation (CMV). HFV has often been combined with or in some cases even completely replaced CMV in the attempt to reduce iatrogenic injury. High frequency percussive ventilation (HFPV) is a specific mode of HFV that has been successfully applied in the treatment of acute respiratory failure after smoke inhalation; it has also been more widely used in pediatric than in adult patients. This article gives an introduction to and a description of the basic principles of HFPV, a mode of ventilation which we found particularly versatile and reliable in our preliminary clinical experience with the maneuver. PMID- 14735025 TI - High frequency percussive ventilation (HFPV). Case reports. AB - Treatment of acute respiratory failure is still a hot issue in intensive care everyday practice: in the last few years high frequency ventilation techniques have been employed as a therapy for adult respiratory distress syndrome (ARDS) and acute respiratory failure (ARF). We applied high frequency percussive ventilation (HFPV) to 3 patients affected by ARDS or ARF, who did not improve after 24 hours of conventional mechanical ventilation (CMV). All our patient underwent 12 hours of HFPV, and showed an improvement of both respiratory exchange and radiological imaging. Even if the pathogenesis of ARF was quite different, in all patient we registered a good response and no complications. PMID- 14735026 TI - An unusual aspect of tracheal stenosis. Case report. AB - lnjury of the tracheal mucosa at the decubitus site of the endotracheal tube cuff during prolonged endotracheal intubation and resulting fibrin deposits may predispose for the development of tracheal stenosis. Frequent endoscopic control examinations, following the increased use of dilation tracheostomy techniques, have revealed a considerable number of these once misrecognized complications in laryngeal and tracheal structures alike. The case reported here appears to confirm this sequence of events. Timely operative endoscopy using a pair of pincers mounted on a rigid endoscope permitted the removal of the fibrin membrane causing the tracheal lumen stenosis and allowed us to achieve complete and definitive recalibration of the trachea, with restoration of spontaneous breathing. Cortisone therapy prolonged for 5 days probably prevented recurrence of the stenosis. Follow-up was carried out in 3 phases. The 1(st) phase included early control using tracheal endoscopy; 2(nd) comprised neck CT scan to examine the tracheal lumen 15 days after endoscopic control, and the 3rd phase involved medical examination after about 3 months and neck radiography in 2 projections. PMID- 14735027 TI - Noninvasive characterization of myocardium after transmyocardial laser revascularization. AB - AIM: The therapeutic mechanism of transmyocardial revascularization (TMR) is not yet fully understood, and continues to be a subject of controversy and active research. Immediate direct laser channel flow, gradual angiogenesis, denervation, and perioperative infarction of the ischemic area have been all discussed, without clear evidence indicating superiority of individual factors. METHODS: We utilized a prospective noninvasive physiologic dynamic method to assess laser related myocardial injury. The study protocol included EKGs and echocardiograms, including intraoperative transesophageal echocardiograms (TEE) on consecutive TMR patients. CPK-MB was measured postoperatively, with 5 samples at 6-hour intervals. RESULTS: Fifty male patients averaging 62 years old were enrolled in the study. Two patients experienced postoperative myocardial infarctions, from which 1 died. The average CPK-MB values were 12.8+/-1.28 immediately after surgery, 19.2+/-2.4 at 6 h, 15.2+/-2.3 at 12 h, 12.2+/-6.3 at 18 h, and 11.7+/ 1.3 at 24 h. In only 5 patients were the CPK-MB values over 30 units at their peak. The intraoperative wall motion remained unchanged in the patients studied, both using TEE and transthoracic echography. CONCLUSION: Significant myocardial injury after TMR appears unlikely, as indicated by CPK-MB and myocardial wall dynamics. Furthermore, TMR does not seem to aggravate baseline myocardial ischemia. We found no evidence to support a hypothesis that surgical myocardial injury constitutes the mechanism of therapeutic action in TMR. PMID- 14735028 TI - Ultrastructural changes of cardiac valves in bacterial endocarditis. AB - AIM: The principal objective of this study was to document morphological changes in valves with acute endocarditis in order to gain further knowledge of the pathogenesis of these diseases. METHODS: Scanning and transmission electron microscopic investigations were carried out on explanted human heart valves to reveal ultrastructural changes due to bacterial endocarditis. RESULTS: Bacterial inflammation endocarditis initially induced metaplasia of the endothelial cells which then lose contact with each other. In the 2nd phase of the disease, the collagen fibres are systematically removed whereby large cavities appear. In the 3rd phase, localised hyperplasia of collagen fibres was observed often resulting in the development of vegetation. The ultrastructural changes are uniform and independent of the bacterial species. CONCLUSION: Bacterial endocarditis is therefore a set of complex interactions between endothelial cells and bacteria which should be taken into consideration for the development of new therapeutic approaches. PMID- 14735029 TI - Twenty-year follow-up of the Carpentier-Edwards standard porcine bioprosthesis in the Oriental population. AB - AIM: The 20-year period long-term results of porcine bioprosthetic valve use are limited. In addition, the majority of these reports come from Western countries. Given the scanty information reported in Oriental countries, this study was therefore designed to examine 20-year long-term results in patients who received a Carpentier-Edwards porcine bioprosthetic valve in an effort to contribute further information on the long-term clinical performance of porcine prosthetic valves from a viewpoint of results in the Oriental population. METHODS: From July 1979 to April 2001, 82 patients received valve replacement with a standard Carpentier-Edwards porcine valve. There were 40 men and 42 women with a mean age of 42.3+/-15.1 years (range 16 to 73 years). Follow-up time extended more than 20 years (mean 10.9+/-3.2 years, range 0.5 to 21.5 years ) for a total of 719.5 patient-years. RESULTS: The overall operative mortality was 16.9% (14 of 83 procedures). At 5, 10, 15, and 20 years, the actuarial survival rate of patients was 71.7%, 66.9%, 55.5%, and 44.4%, respectively. Actuarial estimates of freedom from structural valvular deterioration (SVD) at 5, 10, 15, and 17 years were 96.3%, 64.0%, 24.3%, and 24.3%, respectively; from reoperation 96.3%, 64.5%, 24.5%, and 24.5%; from operated valvular endocarditis 96.8%, 92.6%, 92.6%, and 92.6%; and from overall thromboembolism 96.3%, 88.5%, 67.2%, and 52.2%. In normal sinus rhythm, actuarial estimates of freedom from thromboembolism at 5, 10, 15, and 17 years were 100.0%, 100.0%, 81.8%, and 81.8%, respectively. Whereas for those in patients with atrial fibrillation, the estimates of freedom from thromboembolism were 94.5%, 82.4%, 57.7%, and 38.5%. CONCLUSION: This study demonstrates the very satisfactory 20-year period long-term performance of freedom from bleeding events, thromboembolism (except in patients with atrial fibrillation), and valvular endocarditis in Oriental patients undergoing replacement with a porcine valve. However, the remarkable rate of SVD and reoperation ensued at 6 years after bioprosthesis implanted which does not differ from the series reported from Western countries. PMID- 14735030 TI - Role of age, gender and association of CABG on long-term results after aortic valve replacement with a Carpentier-Edwards bioprosthesis in the elderly. AB - AIM: The problem of postoperative sudden death and the effect of age, gender and association of coronary artery bypass grafting were studied after implantation of a bioprosthetic valve in aortic position. METHODS: DESIGN: retrospective investigation during 13 year. SETTING: general hospital. PATIENTS: 500 mostly symptomatic patients who received this bioprosthesis had a follow-up of 2,022 patient-year for 499 patients. INTERVENTION: aortic valve replacement with a Carpentier-Edwards pericardial prosthesis. MEASURES: hospital complications and mortality, long-term mortality with focus on sudden death and its risk factors, valve related complications and other cardiac events. RESULTS: Within the hospital: atrial fibrillation was the most frequent complication, for which only gender had an effect (p=0.014). The most occurring valve related complications were thrombo-embolic events. Mortality was adversely affected by male gender (p=0.040). Long-term results: thrombo-embolic events were the most important valve related complications. These events and haemorrhage, endocarditis and reoperation rate were not affected by the association of a CABG. Univariate analysis in patients over 73 showed that the need for CABG significantly increased global mortality (p=0.0001), occurrence of cardiac fatality (p=0.0003) and congestive heart failure (p=0.0036). Non-valve related factors seemed most responsible for postoperative sudden death. CONCLUSION: Age, male gender and the association of a CABG remain important determinants for postoperative survival, but not for the occurrence of valve related complications. Sudden death seems not always related to the valve prosthesis, but is as such classified by convention. PMID- 14735031 TI - Neurologic symptoms after great saphenous vein harvesting for coronary artery bypass grafting. AB - AIM: Incidence evaluation of cutaneous neurologic symptoms in the lower limbs as a new event after great saphenous vein (GSV) harvesting for coronary artery bypass grafting (CABG). Each day we harvest the GSV for CABG. Some authors have reported the onset of saphenous neuralgia complex as a new event of which we would evaluate the incidence. METHODS: From January 2000, until June 2001, 2,091 patients underwent cardiac surgery; 1,326 underwent CABG, 1,227 of them using the GSV as a conduit for almost one graft. These patients were prospectively reviewed; all were preoperatively examined to determine the presence of normal sensation in the lower limbs and elude the presence of saphenous neuralgia. Then, we evaluated sensations in the lower limbs at 5 days, 8 weeks, and 5 months after operation to determine the new onset of saphenous neuralgia. The areas of sensory loss were recorded each time and reported in a diagram to obtain 3 areas. RESULTS: Hyperaesthesia and pain were noted in a few patients, especially at 5 days and 8 weeks control, but at 5 months none of them complained of real pain. CONCLUSION: This study demonstrates that saphenous neuralgia after harvesting the GSV for CABG is a rare consequence. The main symptom is anaesthesia but its duration is generally no longer than 2 months. Hyperaesthesia and pain, for the early onset and the early disappearance, are considered as a normal consequence of surgical procedure. PMID- 14735032 TI - The role of posterior pericardiotomy on the incidence of atrial fibrillation after coronary revascularization. AB - AIM: Pericardial effusion and atrial fibrillation (AF) are two common complications in coronary revascularization surgery. The aim of this study was to evaluate the efficiency of posterior pericardiotomy in pericardial effusion and AF. METHODS: This randomized prospective study includes 113 patients who underwent isolated CABG procedure between May 2000 and December 2000 in our hospital. Posterior pericardiotomy incision was done in Group I (n=54). Group II constituted the control group (n=59). Postoperative pericardial effusion was assessed by echocardiography and rhythm follow-up was done by the same cardiologist. RESULTS: There was no significant difference between study group and the control group considering the chest drainage (940.18+/-367.96 vs 894.92+/ 360.65; p=0.507). The number of patients with remarkable intrapericardial effusion (>50 ml) was significantly lower in the posterior pericardiotomy group (25.93% vs 47.45%, p=0.020). The incidence of postoperative AF was no different between the posterior pericardiotomy group and the control group (12.96% vs 20.34%; p=0.32). In both groups, the incidence of AF was significantly higher in patients with mild or moderate pericardial effusion (29%), compared to patients with no or minimal pericardial effusion (10%), (p=0.017). CONCLUSION: Posterior pericardiotomy significantly reduces the pericardial effusion in coronary bypass procedure postoperatively. Patients with pericardial effusion were subjected to AF more frequently. PMID- 14735033 TI - Intrathoracic pulmonary artery catheter allocation in the background of left atrial dilatation. AB - AIM: The aim of the study was to find out whether dilatation of the left atrium (LA) influences the intra-thoracic distribution of thermodilution pulmonary artery (TPA) catheter in either branch of pulmonary artery and compare the measured cardiac output. METHODS: In this prospective study of 132 consecutive patients in a university hospital setting, LA size and ejection fraction was assessed by echocardiography, in the preoperative period. In 66 patients posted for coronary artery bypass grafting (Group 1), a standard anaesthesia regimen was used and TPA catheter was floated through the right internal jugular. In another 66 patients of long-standing mitral stenosis for mitral valve repair/replacement (Group 2), TPA catheters were similarly floated. Intrathoracic placement of the tip of the TPA catheter into the right or left pulmonary artery (PA) was confirmed on chest X-ray. TPA catheter length to its wedging, intra-arterial pressure, heart rate, PA pressure, pulmonary artery wedge pressure (PAWP) and cardiac output by thermodilution technique were noted. RESULTS: Leftwards TPA catheter placement was significantly (p<0.001) more frequent (71%) in mitral stenosis patients (group 2) than the CABG (group 1), (18%). On regrouping the observations of rightwards placed TPA (Group R) and leftwards placed TPA (Group L), we observed that large LA (> or = 25 mm3/m2) body surface area (BSA) and high PAWP (> or = 20 mmHg) was associated with significantly (p<0.001) higher incidence of leftwards TPA catheters. Positive predictive value of both the factors in combination was significantly higher (96%) than individual factors large LA (81%) and high PAWP (88%). CONCLUSION: In long standing mitral stenosis, left atrium enlargement, with high PAWP and the hypokinesia of left atrium (atrial fibrillation) likely to influence the angulation of left PA with main PA and so the predominant entry of TPA catheter tip in left PA. PMID- 14735034 TI - Cardiac surgery in patients receiving long term hemodialysis. Short and long term results. AB - AIM: Cardiac surgery carries a high risk in hemodialysis patients and has been questioned for its results; the purpose of this study is to focus on the short and long term results in our institution. METHODS: We retrospectively analyzed the data from 124 hemodialysis patients who underwent cardiac surgery in our unit between January 1980 and December 1998; 14.5% were diabetic; 46% had isolated coronary artery disease (group 1); 29.8% had valvular disease alone (group 2); 14.5% valve and coronary disease (group 3) and 9.6% miscellaneous disease at highest risk (group 4). We analyzed the relationship between several variables (age, sex, hypertension, diabetes, previous myocardial infarction, type of disease, preoperative ejection fraction) and operative mortality (30 days) and late survival. RESULTS: The overall operative mortality was 16.9%. The only risk factor was the type of cardiac disease: operative mortality was higher in groups 3 and 4 combined than in groups 1 and 2 combined (30% versus 12.7%, p=0.07). Ninety-nine patients were followed until January 2002. Late survival rate was 46.6+/-5% at 6 years for all patients, it was significantly better in groups 1 and 2 combined than in groups 3 and 4 combined. The only risk factor for late mortality was arterial hypertension. Fifty-seven patients are still alive, 46 in groups 1 and 2, 11 in groups 3 and 4. Progression of coronary lesions occurred in 6 patients and valvular lesions in 3 patients. The remainder are doing well. CONCLUSION: Cardiac surgery seems to be justified by the severity of the lesions. Its actual results can perhaps, be improved by earlier detection of cardiac disease and better prevention of myocardial hypertrophy and cardiac calcifications. PMID- 14735035 TI - Cerebral vasoreactivity does not predict cerebral ischaemia during carotid endarterectomy. AB - AIM: Assessment of cerebrovasoreactivity (CVR), obtained by transcranial Doppler (TCD) and the acetazolamide test to predict cases requiring selective carotid shunting on the basis of neurologic monitoring. METHODS: A consecutive series of 87 carotid endarterectomy (CEA) cases was studied. Before surgery CVR was evaluated by measuring the mean velocity of the middle cerebral artery (mvMCA) using TCD at the basal condition and at 30 min after intravenous administration of acetazolamide (1 g). Carotid shunting was performed using neurologic monitoring under local anesthesia. Receiver operating characteristic (ROC) curve was calculated for sensitivity and specificity for various CVR cut offs. RESULTS: The ROC curve demonstrated that there was no single CVR cut off with both sensitivity and specificity above 80%. CONCLUSION: The present study, which employed neurologic monitoring as the method of comparison, did not prove that CVR, as calculated by TCD and the acetazolamide test, is a valid preoperative test for predicting cerebral ischaemia caused by carotid clamping. PMID- 14735036 TI - Combined minimally invasive procedure for descending thoracic aortic aneurysm with left main coronary artery disease. AB - A 78-year-old man had a descending thoracic aortic aneurysm with left main coronary artery disease. Combined beating heart coronary artery bypass and endovascular thoracic aortic aneurysm repair was performed without cardiopulmonary bypass. The left anterior descending artery and the obtuse marginal branch of the left circumflex artery were revascularized through a left antero-lateral small thoracotomy. The aneurysm was excluded with stent grafts through a small femoral incision. This combined less invasive procedure is a promising approach providing better surgical results in patients with thoracic aortic aneurysm and severe coronary artery disease. PMID- 14735037 TI - True acquired left ventricular aneurysms in childhood. AB - AIM: True left ventricular aneurysms are rarely encountered in childhood. Etiological factors are varied and often indeterminable. There is a paucity of literature detailing the surgical techniques used to repair pediatric aneurysms. METHODS: Three children aged 21 months, 19 months and 5 years with true left ventricular aneurysms underwent surgical repair in our unit. The clinical details, surgical technique, histological review, hospital outcome and short term follow-up is discussed. RESULTS: Two patients underwent repair of their aneurysms by means of the Cooley endoaneurysmorraphy technique, and 1 patient underwent repair using the aneurysm resection and linear plication technique. Histological examination revealed tuberculosis to be the etiological factor in one patient, while a non-specific vasculitis was present in the other 2 patients. Immediate postoperative course was uneventful and all 3 patients were discharged from hospital within one week. Two patients were recatheterized during follow-up visits. All patients were asymptomatic at 1 year follow-up. Echocardiography demonstrated good left ventricular function with disappearance of preoperative mitral regurgitation. CONCLUSION: Pediatric left ventricular aneurysms may be successfully repaired using standard operative techniques. These techniques render excellent short-term RESULTS: Longer follow-up is needed to fully evaluate the outcome of these children. PMID- 14735038 TI - Cardiac tamponade caused by an ingested sewing needle. A case report. AB - Ingested sharp-pointed foreign bodies can cause serious complications. A case of a 16-year-old girl with cardiac tamponade due to ingestion of a sewing needle is presented. Ingested needles have often been reported as a cause of gastrointestinal injuries but in this rare case the sewing needle actually migrated into the myocardium. PMID- 14735039 TI - A new technique for abdominal heart transplantation in rats. AB - Experimental studies in animals play a major role in the progress of medicine. Different surgical techniques have been described for heterotopic heart transplantation in rats. In this study we introduce a new technique for heart transplantation in the abdominal cavity of rats. Fifteen Sprague Dawley rats have been used as recipients and 15 others as donors. Following preparation of recipient abdominal aorta and left renal vein, the donor heart including proximal arcus aorta was harvested. Donor aorta was anastomosed to the recipient's aorta; donor pulmonary artery was anastomosed to the left renal vein of the recipient using continuous suture technique. Graft function was evaluated daily by palpation of the rat abdomen. The mean operating time was 38.46+/-2.66 min and the mean ischemia time was 23.93+/-2.11 min. One death was seen because of bleeding of the aorto-aortic anastomosis. In this study we evaluated advantages and disadvantages of our technique and compared it to other techniques. This modification provides a more anatomical position, reduces exploration time, has a low incidence of morbidity and mortality. We conclude that, this implantation technique is more suitable for heterotopic heart transplantation in rats. PMID- 14735040 TI - Clinical results of aortic arch replacement using a four branched prosthetic graft. AB - AIM: The aim of this study was to evaluate the operative techniques of total arch replacement, the clinical results and the survival curves of patients following this procedure. METHODS: Since December 2001, 92 patients have undergone surgical treatment for aortic dissection and aneurysm. The total aortic arch replacement was performed in 24 of these patients. There were 16 men and 8 women, and the age range was 42 to 81 years with a mean age of 59.4 years. As the operative technique for total arch replacement, we used the 4-branched prosthetic graft, selective cerebral perfusion (SCP), continuous cold blood cardioplegia (CCBC), and open distal anastomosis under circulatory arrest. The combined operations were coronary bypass grafting in 4 patients, aortic valve suspension in 1 patient and a Bentairs procedure in 1 patient. Eleven (73.3%) patients with acute dissection required emergency operation. RESULTS: The hospital mortality rate was 25% (6 of the 24 patients). The causes of death were multiple organ failure (MOF) due to renal and mesenteric ischemia in 3 patients, cerebral infarction in 2 patients, myonephropathic metabolic syndrome (MNMS) in 1 patient, respectively. The data concerning extracorporeal circulation was 204+/-53 min in total pump time, 136+/-43 min in aortic cross clamp time, 83+/-14 min in SCP time and 48+/ 10 min in circulatory arrest time, respectively. The long-term result in actuarial survival rate was 76% for 5 years. CONCLUSION: We consider the technique of total arch replacement using 4-branched prosthetic graft, SCP, CCBC, and open distal anastomosis is a useful operative method in patients with aortic aneurysm. PMID- 14735042 TI - Endovascular repair of descending thoracic aortic aneurysm via midsternotomy and without extracorporeal circulation. AB - A 68-year-old man with peripheral vascular disease and associated risk factors, was diagnosed by CT-scan and aortography with an atherosclerotic descending thoracic aortic aneurysm. The patient was treated by successful endovascular grafting of the aneurysm using a midsternotomy incision and the insertion of a thoracic stent-graft through a T-Dacron tube sutured in the ascending aorta, without the aid of cardiopulmonary bypass. PMID- 14735041 TI - Topical negative pressure therapy. A very useful new method to treat severe infected vascular approaches in the groin. AB - AIM: The treatment of infected vascular surgery sites is challenging. Negative pressure applied uniformly to the entire wound surface has been shown to allow granulation tissue formation and to promote healing of acute and chronic wounds. METHODS: We used the Vacuum-Assisted Closure (VAC, Kinetic Concepts Incorporated, San Antonio, Texas, USA) system in 4 patients with severe groin wound infection after emergency surgery on the femoral artery. RESULTS: In all 4 patients, general health improved and the wound changed rapidly from a large infected cavity to a minor lesion readily covered using a simple surgical technique. CONCLUSION: This study establishes VAC as a very valuable tool for managing severe complications of groin vascular surgery sites even in patients with obesity and/or diabetes mellitus. PMID- 14735043 TI - Cystic lesions of the pericardium. Review of the literature and classification. AB - Pericardial cystic lesions (PCLs) occur infrequently but are significant for their varying clinical presentation and pathological multitude. A review of the literature (including Medline and Current Contents database searches, and search of existing bibliographies) finds confusion in nomenclature and an absence of appropriate classification. A new classification system is proposed based on exo- or endophytic growth, presence of adhesions, and compression of myocardium or great vessels. A multitude of pathological entities with diverse pathogenesis, disease courses, and prognoses may present as PCLs. Detailed knowledge of lesion types and alternatives among diagnostic and therapeutic options permits a selective approach to patient management. The usefulness of a unified classification system should be evaluated in a substantial patient population, with detailed statistical analysis. PMID- 14735044 TI - Apoptotic cell death and genetic control in graft coronary artery disease in heart transplant. AB - AIM: Apoptosis is a type of programmed cell death whereby, immunologic, genetic and biochemical mechanisms are involved in its control. On the other hand, graft coronary artery disease is the most important restrictive factor for the long term survival of heart transplantation. The purpose of this study is to analyse both apoptotic cell lesions in transplanted patients that present coronary artery disease. METHODS: From August 1984 until December 1996, 148 heart transplants were carried out in the Clinica Universitaria de Navarra. In 102 patients, annual coronary angiography was performed, reaching a diagnosis of coronary artery disease in 30 patients. Study of apoptotic cell death was done in the tissue of endomyocardial biopsies on all patients by means of the TUNEL technique. Procedures of immunohistochemistry with antibodies antic-myc, p53 and bcl-2 were carried out and results were compared with a control group of 30 patients with homogeneous characteristics. RESULTS: All patients with coronary artery disease showed apoptotic cardiomyocytes, 13 patients to a mild degree, 14 to a moderate degree and 3 to a severe degree, while in the control group apoptosis was found only to a mild degree in 8 patients, obtaining a very significant statistical difference (p<0.0001). The expression of analysed oncoproteins was null in the 2 groups. CONCLUSION: Myocardial apoptosis is a constant finding in transplanted patients with coronary artery disease. We have not seen any correlation between the apoptotic process and genetic mechanisms. PMID- 14735045 TI - Is early hospital discharge feasible following normothermic coronary artery surgery on the fibrillating heart? AB - AIM: Protocols for the earlier discharge of cardiac surgical patients are gaining popularity. We present our experience with an early hospital discharge policy following coronary artery bypass grafting (CABG) on the fibrillating heart. METHODS: Three-hundred and ninety-two consecutive patients who underwent elective CABG by a single surgeon were retrospectively reviewed. CABG was performed initially (1998-1999) in 191 patients with cardiopulmonary bypass (CPB) normothermia, intermittent aortic cross-clamping (AXC) and ventricular fibrillation and later (2001-2003) in 201 patients without CPB. Emphasis was given on short AXC and CPB times, early extubation, early mobilization and atrial fibrillation prophylaxis. Discharge criteria were as follows: walking on stairs unassisted, sinus rhythm for 24 hours, normal bowel function, apyrexia, family support at home. A 6-week follow-up clinic visit was arranged. Hospital re admissions were carefully monitored. RESULTS: The mean (+/-SD) age of the patients was 62+/-9.6 years and the mean Parsonnet score was 6.7. The mean hospital stay was 6.1+/-2.5 days. Sixty-three (16%) and 171 (44%) patients were discharged by postoperative day 4 and 5, respectively. The following factors were independently associated with longer hospital stay: number of grafts performed (>3), requirement for postoperative inotropic support and social circumstances inadequate for early discharge. Twenty-three patients (5.8%) were re-admitted in the 6-week postoperative period. Shorter hospital stay was not associated with increased risk of re-admission. CONCLUSION: Early discharge after CABG with ventricular fibrillation is achievable, comparable to "fast-track techniques" without the use of CPB and is not associated with higher re-admission rates. We recommend the routine use of this protocol in all patients undergoing primary elective CABG. PMID- 14735046 TI - Midterm experience with the Sorin Bicarbon heart valve prosthesis for rheumatic disease. AB - BACKGROUND: In this study, we present a single center experience with Bicarbon bileaflet valve in 307 patients with rheumatic heart disease. METHODS: Between August 1998 and September 2000, 307 patients underwent heart valve replacement using the Bicarbon bileaflet valve (Sorin Biomedica, Saluggia, Italy) with an average age of 47.19+/-13.21 years (range 14 to 80 years), consisting of 147 males and 160 females at Alkan Hospital, Cardiovascular Surgery Department. Aortic valve replacement (AVR) was performed in 77 patients, mitral valve replacement (MVR) in 156 patients and double valve replacement (DVR) in 74 patients. RESULTS: The early mortality rate was 3.3% (10/307) and there was no late mortality. The actuarial survival rate, including hospital mortality, was 96.74+/-1.01% for the whole group, 96.5+/-1.5% for the MVR group, 97.4+/-1.8% for the AVR group and 97.3+/-1.9% for the DVR group at 35 months. One patient had obstructive valve thrombosis with MVR. The 35 months actuarial freedom from valve thrombosis was 99.58+/-0.4% for the whole group. Four patients were reoperated and the 35 months actuarial freedom from reoperation was 98.53+/-0.7% for the whole group, 98.65+/-0.9% for the MVR group, 96.73+/-02% for the DVR group and 100% for the AVR group. No instances of perivalvular leak, hemolysis, endocarditis or embolism were observed during the entire follow-up period. Mean follow-up duration was 16.5+/-7.9 months (ranged 4 to 35 months). CONCLUSION: We have presented our mid-term results with the Sorin Bicarbon bileaflet valve in patients with rheumatic heart disease, which provided good clinical performance combined with meticulous patient care and advanced surgical techniques. PMID- 14735047 TI - Helicobacter pylori seropositivity in patients with unstable angina. AB - AIM: The pathogenesis of ischemic heart diseases has been correlated, on epidemiological and pathogenetic grounds, with infections by viruses and bacteria, including Helicobacter pylori (H. pylori). THE AIM: of this study were to investigate the association of unstable angina (UA) with anti-H. pylori seropositivity in a case-control study and to search for the classic cardiovascular risk factors in both infected and uninfected patients. METHODS: We studied 32 consecutive patients (20 males, 12 females), mean age 65 years (range 42-89), with final diagnosis of UA. A total of 64 subjects (40 males, 24 females, mean age 65 years, range 42-89) admitted to the Emergency Care Unit, age and sex matched, served as controls. The presence of hypertension, serum levels of cholesterol and glucose, plasma levels of fibrinogen, smoking habit and social class were investigated in all patients. Cases and controls were inhabitants of NorthWestern Italy, and had similar socioeconomic status as based on working place and on instruction level. H. pylori seroprevalence was assessed by the presence of antibodies (IgG) against H. pylori by means of a commercial enzyme immunosorbent assay. RESULTS: Antibodies to H. pylori were found in 26/32 (81%) of the patients and in 34/64 (53%) of the controls (p=0.007); the odds ratio was 3.82 (95% confidence interval 1.27 to 12.04). Classical cardiovascular risk factors, such as socio-economic status, did not differ among patients with and without antibodies to H. pylori. CONCLUSION: Patients with unstable angina had a significantly higher seroprevalence of anti-H. pylori than the control population. Classical risk factors for ischemic heart disease, such as the indicators of socio-economic status, were equally distributed among infected or uninfected patients with UA. PMID- 14735048 TI - The effects of heparin coated circuits on pulmonary injury. A clinical study. AB - AIM: Heparin-coated circuits have dramatic effects on the coagulation cascade, but their role on complement activation has not been clearly defined. In this clinical study the effect of heparin-coated circuits on static lung compliance and pulmonary vascular resistance is described. METHODS: Thirty patients were randomly divided into two groups: with either a heparin-coated circuit or an identical but non-coated circuit control group. In the heparin-coated group, all the blood contacting surfaces were treated with immobilized heparin (Duraflo II.) RESULTS: Early postoperative pulmonary function is determined with measurements of static lung compliance, pulmonary vascular resistance and arterial blood gases. Static lung compliance was significantly better in the heparin coated (HC) group in the early postoperative period (p=0.001). Pulmonary vascular resistance was significantly lower in the heparin-coated (HC) group in the early postoperative period (p=0.001). CONCLUSION: We believe that the method of heparin binding may play a role in its diminished effect on complement activation, but the general augmentation of the circuit's biocompatibility may explain its beneficial effect on pulmonary vascular resistance and static lung compliance. PMID- 14735049 TI - Coronary artery bypass grafting in dextrocardia with situs inversus totalis. AB - Dextrocardia with situs inversus totalis is a rare congenital abnormality of development involving a left-handed malrotation of the visceral organs. The incidence of coronary artery disease in this condition is probably similar to that in the general population. We report herein the case of a 62-year-old man with dextrocardia and situs inversus totalis who underwent myocardial revascularization by the use of right internal mammary artery and radial artery. PMID- 14735050 TI - Undifferentiated sarcoma of the mitral valve with unique clinicopathologic presentation. AB - We report a case of undifferentiated sarcoma originating from the mitral valve in a 3-year-old boy who presented with embolic symptoms of the lower extremity. The tumor was demonstrated by echocardiography. The mass was removed under a mild hypothermic cardiopulmonary bypass with cold blood cardioplegia. Grossly, the mass was composed of multiple small nodules, and the histopathologic findings were compatible with an undifferentiated sarcoma. Postoperative chemotherapy was added. The patient remains without evidence of recurrence 18 months after surgery. PMID- 14735051 TI - An unusual cause of hemoptysis: ventriculopulmonary fistula. AB - A 55-year-old man presented with massive hemoptysis following coronary artery bypass grafting and repair of a left ventricular aneurysm. Radiological and bronchoscopic examinations revealed no bronchial cause. The findings of computed tomography (CT) of the chest and echocardiography showed a pseudoaneurysm of the left ventricle. Surgical exploration confirmed that the pseudoaneurysm communicated with the lung parenchyma. PMID- 14735052 TI - Mininvasive abdominal aortic surgery. Early recovery and reduced hospitalization after multidisciplinary approach. AB - AIM: Clinical experience in gastrointestinal surgery demonstrated that a multimodal approach can improve the outcome and reduce the length of hospital stay. In this paper we investigate the impact of a multimodal clinical program, based on mininvasive surgery, epidural anesthesia and early feeding and mobilization, on postoperative morbidity and hospitalization after abdominal aortic surgery. METHODS: A 2-armed study was designed. All patients undergoing abdominal aortic surgery between May 2000 and April 2001 were enrolled in a multidisciplinary clinical program including thoracic epidural anesthesia and analgesia, left sub-costal minilaparotomy without evisceration, encouragement to feed and mobilize soon after surgery (Multidisciplinary group: n=82). For comparison purposes, a retrospective analysis was conducted using the data of all patients operated on between January and December 1997, receiving standard anesthesia care and a standard surgical and nursing program (Standard group: n=64). RESULTS: In the Multidisciplinary group we observed significantly better pain relief (p<0.01), earlier restoration of ambulation (p<0.01), earlier feeding (p<0.01) and passage of stools (p<0.01). The incidence of complications was significantly lower in the Multidisciplinary group: pulmonary (0% vs 14.1%), cardiac (2.4% vs 9.4% ) and gastrointestinal (0% vs 10.9%). None of the patients in the Multidisciplinary group required admission to Intensive Care. Median postoperative hospitalization was 3 days in the Multidisciplinary group compared to 9 days in the Standard group (p<0.01). CONCLUSION: These results suggest that a multidisciplinary intervention with review of the traditional surgical care program would enhance recovery, decrease morbidity and hospitalization after abdominal aortic surgery. PMID- 14735053 TI - Blood distribution to the anterior spinal artery from each segment of intercostal and lumbar arteries. AB - AIM: Prevention of paraplegia, a serious complication of surgery for thoracoabdominal aortic aneurysm, has been well documented. However no assured prophylaxis against this complication has yet been found. Spinal ischemia is believed to be the major cause of paraplegia. We conducted an experimental study to define the development of paraplegia with regard to the blood supply to the spinal cord. METHODS: A porcine model was used to evaluate blood distribution to the anterior spinal artery. Colored silastic agent was selectively injected into the intercostal and lumbar arteries, and distribution to the anterior spinal artery was evaluated on 50 animals. The intercostal and lumbar arteries were ligated in the segments where the blood supply to the anterior spinal artery would be interrupted. Whether or not paraplegia developed was checked 2 days later. RESULTS: Colored silastic agent arrived at the anterior spinal artery from all segments of the 8th intercostal to 4th lumbar arteries. Two of 9 pigs (22.2%) that underwent ligation of the segments from the 9th intercostal to 2(nd) lumbar artery suffered paraplegia. In 3 non-paraplegic pigs, colored silastic agent injected into the preserved arteries was found to have covered a wider range. CONCLUSION: All the intercostal and lumbar arteries supplied blood to the anterior spinal artery. When large segments of intercostal and lumbar arteries were ligated, the blood flow from the preserved segments acquired increased dominance. The possibility exists that any intercostal and lumbar artery can supply blood to the spinal cord and become collateral circulation to the anterior spinal artery. PMID- 14735054 TI - Aneurysms of the hypogastric artery following surgery of the abdominal aorta. A report of two cases and review of the literature. AB - Isolated aneurysms of the hypogastric artery are very rare and account for between 0.04% and 0.4% of all intra-abdominal aneurysms. In 85% of cases they are monolateral and are present in association with an aneurysm of the infrarenal abdominal aorta and of the common and external iliac arteries, they make up part of a poly-aneurysmal disease. Unless the patient is an extremely poor condition, surgical treatment is generally indicated for aneurysms greater than 3 cm; close monitoring of those with smaller aneurysms is recommended. Two patients presented with isolated aneurysm of an internal iliac artery which had developed several years after aortoiliac surgery. The one received surgical treatment; the other, who was in poor general conditions and at high risk for surgery, underwent endovascular embolization. Both procedures were successful, with a current follow up between 15 and 18 months. Endovascular embolization, as performed in the 2(nd) patient, provided an alternative to the surgical procedure. After injection in the aneurysmal sac of the Gianturco spirals, a covered stent was placed in the iliac axis to exclude the inflow of the hypogastric artery. According to our experience of 2 patients, the one treated surgically and the other submitted to a less invasive endovascular procedure, we can state that both methods are practicable. The final choice lies with the vascular surgeon, after weighing the multiple factors that each case involves. PMID- 14735055 TI - Descending necrotizing mediastinitis. Diagnosis and surgical treatment. AB - AIM: Descending necrotizing mediastinitis (DNM) is an unusual and severe disease with a high mortality rate. Surgical management remains controversial. Our investigations reviews the most effective surgical treatment in the management of this rare pathology. METHODS: Seven patients with DNM and treated over a 20-year period are reported. All patients were evaluated according to the classification suggested by Endo et al. of the degree of mediastinal diffusion, based on CT scan findings. Five patients underwent combined cervical drainage and thoracotomy, 2 patients were treated with cervical drainage alone. RESULTS: The outcome was favorable in 5 patients, 4 treated with a combined cervical and thoracic approach and 1 with a cervical approach alone. Two patients that underwent a combinated cervical and thoracic approach alone, died of septic shock. Overall mortality rate was 28.5%. CONCLUSION: Early diagnosis and early, aggressive surgical treatment are required to improve the poor prognosis of DNM. Although a unique surgical management is still not completely accepted, we state, in agreement with other authors, a wide approach consisting of a cervical drainage and mediastinotomy in case of upper mediastinitis and a combined cervical and thoracic approach in case of lower mediastinitis. In the course of thoracotomy a wide excision of necrotic and particularly fat mediastinal tissue is needed, to avoid a recurrent infection. A continuous cervico-mediastinal irrigation system is suggested during the postoperative period. PMID- 14735056 TI - Study of the diagnostic difference between the clinical diagnostic criteria and results of immunohistochemical staining of multiple primary lung cancers. AB - AIM: When multiple synchronous or metachronous lung cancer lesions are identified, discrimination of multicentric lung cancers from intrapulmonary metastases by clinical findings is often difficult. When tissue types have the same pathological features, such as combinations of squamous cell carcinoma (SCC), adenocarcinoma (AD) or bronchiolo-alveolar cell carcinoma (BAC), it is especially difficult to distinguish a 2(nd) primary lung cancer from a metastatic lesion. A new strategy for accurate diagnosis of multiple synchronous or metachronous lung cancer is needed because of the difficulty of histological discrimination. METHODS: Of 363 patients with primary lung cancer for which surgeries were conducted at our hospital, 7 cases were diagnosed as synchronous multiple lung cancer (BAC-BAC in 4 cases and SCC-BAC in 3 cases) and 8 cases (BAC BAC in 2 cases, AD-BAC in 1 case, AD-AD in 1 case, SCC-AD in 1 case and SCC-SCC in 3 cases) were diagnosed as metachronous multiple lung cancer according to the clinical diagnostic criteria. This study focused on 8 cases with the combinations AD-AD, AD-BAC, or BAC-BAC. For immunohistochemical staining, we used the antibodies to 6 antigens as follows: CK-19, p53, CEA, Hup-1, PE-10, and Ki-67. RESULTS: Of 4 cases diagnosed as synchronous lung cancer according to the clinical diagnostic criteria, differing immunohistochemical stained images of the lesions were observed in 3 cases, while in the 4th case almost identical immunohistochemical stained images were obtained, which indicated the 2 lesions were the primary and metastatic focuses. Of 4 cases diagnosed as metachronous lung cancer according to the clinical diagnostic criteria, almost identical stained images were seen in 3 cases, which indicated the 2 lesions were the primary and metastatic focuses. CONCLUSION: In general, Type A and Type B in Noguchi's BAC classification, tended to be multiple synchronous or metachronous lung cancer lesions, while AD and Type C in Noguchi's BAC classification tended to be the metastatic focus. For the focuses with tissue type of BAC-BAC, the staining using CK-19, PE-10, and Ki-67 was useful in distinguishing multiple primary lung cancer from pulmonary metastasis in cases with a combination of AD and BAC. PMID- 14735057 TI - The difficult approach to neoplastic superior vena cava syndrome: surgical option. AB - AIM: Superior vena cava syndrome is a dramatic event that can be cured in specialized centers. METHODS: Between 1989 and 1995 6 patients with superior vena cava syndrome underwent surgical treatment for thoracic tumors. In all cases the vena was restricted by a neoplastic sleeve. A median sternotomy was performed in all cases. Two patients received an associated right anterolateral thoracotomy to obtain good surgical exposure for tumor resection and grafting. A 12 mm diameter polytetrafluoroethylene graft was inserted in all cases. The tumor resection was radical in 4 cases (2 thymic carcinomas, 2 malignant germ cell tumors) and palliative in 2 (1 non-small cell lung cancer and 1 mediastinal fibrosis). RESULTS: We had no in-hospital mortality. All patients had immediate relief of obstruction after by-pass. Three patients were alive without disease at the end of follow-up (40-96 mo), one patient died of postoperative complications after 4 mo, 2 patients died of disease after 4 and 12 mo. CONCLUSION: PTFE by-pass graft for treatment of the obstructed SVC relieves SVC syndrome and has good medium term patency. PMID- 14735058 TI - Congenital ventricular aneurysm. A rare case of cardiac tamponade. PMID- 14735059 TI - Coronary revascularization on the beating heart and no-touch technique in patients with porcelain aorta. PMID- 14735060 TI - Pulmonary mature teratoma. Primary? Or metastatic? PMID- 14735061 TI - Vasospastic persons exhibit differential expression of ABC-transport proteins. AB - PURPOSE: To quantify the gene expression levels of the ABC-proteins MDR1 (P glycoprotein) and MRP (multidrug resistance-associated protein) isoforms in isolated mononuclear cells of vasospastic persons with increased Endothelin-1 plasma levels. METHODS: Quantitative real-time RT-PCR was performed to determine the expression levels of the MDR1 (P-glycoprotein) gene and MRP1 to MRP5 genes as well as the expression of the ETA and ETB receptor in mononuclear cells derived from 11 vasospastic subjects compared to 10 healthy controls. RESULTS: Mononuclear cells of vasospastic subjects showed a significant decrease in the expression of MDR1 (P-glycoprotein) gene (p=0.029), MRP2 gene (p=0.003), and MRP5 gene (p=0.013) when compared to healthy controls. These effects were poorly correlated with ET-1 plasma levels. No significant ETA and ETB receptor expression was observed in both groups. CONCLUSIONS: Vasospastic persons differ in their expression pattern of MDR proteins from healthy controls. This might be an indirect effect of elevated ET-1 levels. PMID- 14735062 TI - Choroidal neovascularization: a wound healing perspective. AB - The process of submacular angiogenesis seen in association with a variety of chorioretinal disorders is termed choroidal neovascularization (CNV). It invariably results in significant and permanent vision loss arising from the development of scar tissue formation. At the cellular level, CNV appears to be a component of several key processes that can be broadly referred to as wound healing or tissue repair. Wound healing involves a coordinated cascade of cellular events driven, in the main, by the production of cytokines and which are interpreted by target cells in the context of a continually evolving extracellular matrix (ECM). A similar process occurs in what is clinically termed CNV. Angiogenesis is just one component of this wound healing process. Other key components include inflammation, matrix deposition and remodelling. Thus, in the context of a tissue repair response, viable treatment options for CNV could include therapies other than those that are currently directed at the angiogenic component of this process. PMID- 14735064 TI - Novel mutations of the carbohydrate sulfotransferase-6 (CHST6) gene causing macular corneal dystrophy in India. AB - PURPOSE: Macular corneal dystrophy (MCD) is an autosomal recessive disorder characterized by progressive central haze, confluent punctate opacities and abnormal deposits in the cornea. It is caused by mutations in the carbohydrate sulfotransferase-6 (CHST6) gene, encoding corneal N-acetyl glucosamine-6-O sulfotransferase (C-GlcNAc-6-ST). We screened the CHST6 gene for mutations in Indian families with MCD, in order to determine the range of pathogenic mutations. METHODS: Genomic DNA was isolated from peripheral blood leukocytes of patients with MCD and normal controls. The coding regions of the CHST6 gene were amplified using three pairs of primers and amplified products were directly sequenced. RESULTS: We identified 22 (5 nonsense, 5 frameshift, 2 insertion, and 10 missense) mutations in 36 patients from 31 families with MCD, supporting the conclusion that loss of function of this gene is responsible for this corneal disease. Seventeen of these mutations are novel. CONCLUSIONS: These data highlight the allelic heterogeneity of macular corneal dystrophy in Indian patients. PMID- 14735063 TI - Does lens intrinsic membrane protein MP19 contain a membrane-targeting signal? AB - PURPOSE: Lens intrinsic membrane protein MP19 is the second most abundant major protein of the lens fiber cell membrane and appears to be specific to the lens. Different mutations of this protein are known to cause cataract in both humans and mice. To date, the function of MP19 in the lens is not known, nor is the mechanism by which the protein migrates to the cell membrane. The goal of this study was to determine whether or not MP19 distributes to the cell membrane directed by a peptide signal within the sequence of the molecule. METHODS: Using PCR, MP19 cDNA was truncated to yield separate fragments coding for the first 25, 36, and 64 amino acids of the MP19 polypeptide chain. These PCR fragments were further cloned into mammalian expression vector pcDNA4/TO, a tetracycline regulated vector that, upon induction with tetracycline, allows expression of cDNA inserts within the vector. These vectors expressed each of the MP19 truncated fragments fused to EGFP. Each of the prepared plasmids was transfected into T-REx-293 cells using FuGene 6. Cloned cell lines from each of these transfections were obtained and used in the studies. The fluorescent expressed protein was viewed using confocal microscopy. Proteins from the different cell lines were isolated by different membrane extraction methods and western blot analysis was carried out to further determine the localization of expressed MP19 and MP19 truncated fragments. RESULTS: Cell lines expressing intact MP19/EGFP (with EGFP fused to the COOH-terminal end of MP19, MP19G) fusion protein were observed to traffic MP19 to the cell membrane, where it appeared to sequester in rather large pools. All of the MP19 truncations (with EGFP fused to the COOH terminal end of each truncation; MP19-25G, MP19-36G, and MP19-64G) appeared to also traffic EGFP to the cell membrane. MP19-25G and MP19-36G did not distribute uniformly on the membrane, but appeared to localize into smaller, punctate "spots" of fluorescent material. MP19-64G distributed on the membrane similarly to MP19-25G and MP19-36G, however, the punctate areas of fluorescent material were considerably larger and similar to that demonstrated by intact MP19G. Western blot analysis of isolated total membranes, intrinsic membranes, and lipid rafts showed that MP19G and MP19-64G were associated with the intrinsic membrane fraction while MP19-25G and MP19-36G were at least 75% associated with the intrinsic membrane fraction. All of the preparations appeared to be at least 50% associated with membrane lipid rafts. However, when EGFP/MP19-25 and EGFP/MP19-36 (with EGFP fused to the NH2-terminal end of the truncated peptide, GMP19-25 or GMP19-36) were expressed, the fusion protein was observed to remain completely soluble in the cytoplasm, identical to expressed EGFP alone. Western blots of these two fusion proteins also indicated that the product did not associate with the cell membrane. In contrast, when EGFP/MP19 (with EGFP fused to the NH2 terminal end of intact MP19, GMP19) was expressed, the fusion protein did integrate into the cell membrane, identical to MP19G. Western blot analysis revealed that GMP19 also associated with lipid rafts, identical to intact MP19G. CONCLUSIONS: It appears that the first 25 amino acids of the MP19 molecule are sufficient to target the protein to the cell membrane, and apparently integrate into the membrane. With the addition of more amino acids, the polypeptide distributes in the membrane similarly to that of the intact MP19 molecule. It appears that the first 25 amino acids of the MP19 molecule is, indeed, a membrane signal and integration sequence. Also, at least part of these 25 amino acids must integrate into the cell membrane, but not extend through the cell membrane. PMID- 14735065 TI - Shoulder arthroplasty in patients with osteoarthritis and dysplastic glenoid morphology. AB - Fifteen shoulders with primary osteoarthritis and dysplastic glenoid morphology underwent shoulder arthroplasty (eleven total shoulder arthroplasties and four hemiarthroplasties). Patients were evaluated at a mean of 37 months after replacement with clinical examination, Constant score, subjective patient opinion, and radiographic examination. One glenoid component was loose, necessitating removal; the remainder of the prostheses were in place at latest follow-up. Significant improvement was observed in Constant score (32 points preoperatively vs 71 points postoperatively) and active mobility (anterior elevation, 89 degrees preoperatively vs 146 degrees postoperatively; external rotation, 7 degrees preoperatively vs 45 degrees postoperatively) after arthroplasty. This study demonstrates that the rare scenario of osteoarthritis coupled with a dysplastic glenoid can be treated successfully with shoulder arthroplasty. PMID- 14735066 TI - Anterosuperior impingement of the shoulder as a result of pulley lesions: a prospective arthroscopic study. AB - Lesions of the biceps pulley and the rotator cuff have been reported to be associated with an internal anterosuperior impingement (ASI) of the shoulder. The purpose of this study was to determine the factors influencing the development of an ASI. Eighty-nine patients with an arthroscopically diagnosed pulley lesion were prospectively included in this study. Four patterns of intraarticular lesions could be identified. Twenty-six patients (group 1) showed an isolated lesion of the superior glenohumeral ligament (SGHL). In 21 patients (group 2) an SGHL lesion and a partial articular-side supraspinatus tendon tear were found. Twenty-two patients (group 3) had an SGHL lesion and a deep surface tear of the subscapularis tendon, and in twenty patients (group 4) a lesion of the SGHL combined with a partial articular-side supraspinatus and subscapularis tendon tear was diagnosed. Of the patients, 80 (89.9%) showed involvement of the long head of the biceps tendon including synovitis, subluxation, dislocation, and partial or complete tearing. In 43.8% of all patients, ASI was observed. Whereas ASI was seen in 26.6% and 19.1% of patients in groups 1 and 2, respectively, 59.1% of patients in group 3 and 75% of patients in group 4 were found to have an ASI. ASI was significantly more often seen in patients with additional partial articular-side subscapularis tendon tears (P <.0001). In patients with acromioclavicular (AC) arthritis, ASI (62.5%) was more frequently observed than in patients without AC arthritis (P =.0309). In the multivariate analysis the stepwise selection procedure revealed only AC arthritis and the deep surface tear of the subscapularis (groups 3 and 4) to be significant influencing factors for an ASI. Our findings indicate that a progressive lesion of the pulley system, including partial tears of the subscapularis and supraspinatus tendons, contributes significantly to the development of an ASI. A pulley lesion leads to instability of the long head of the biceps tendon, causing increased passive anterior translation and upward migration of the humeral head, resulting in an ASI. In addition, a partial articular-side subscapularis and supraspinatus tendon tear reinforces the ASI. PMID- 14735067 TI - Treatment of deep infections of the shoulder with pedicled myocutaneous flaps. AB - Infection after reconstruction of the shoulder can lead to significant morbidity and possible destruction of the glenohumeral joint. Often, seemingly small wound problems mask underlying soft-tissue deficiency and instability. We have reviewed our experience with deep infections of the shoulder that have been treated with pedicled myocutaneous flaps. Patients in this series had been treated by a variety of local measures including debridement and attempts at reclosure, often unsuccessfully. The ability to import well-vascularized tissue that can treat dead space as well as reestablish a new cutaneous envelope around the shoulder is desirable and indicated for these difficult cases. Surprisingly, even with exposure of the humeral head in the wound, the shoulder joint can be salvaged by use of the techniques described in this article. PMID- 14735068 TI - An assessment of the interexaminer reliability of tests for shoulder instability. AB - Accurate noninvasive clinical tests of shoulder instability are important in assessing and planning treatment for glenohumeral joint instability. An interexaminer agreement trial was undertaken to estimate the reliability of commonly used clinical tests for shoulder instability. Thirteen patients with a history suggestive of instability, who had been referred to a shoulder specialist for treatment of their symptomatic shoulders, were examined by four examiners of differing experience. Good to excellent interexaminer agreement was found for most variations of the load-and-shift test, with the best agreement in the 90 degrees abducted position for the anterior direction (intraclass correlation coefficient [ICC] = 0.72) and in the 0 degrees abducted position for the posterior (ICC = 0.68) and inferior (ICC = 0.79) directions. Fair to good interexaminer reliability was found for the sulcus sign (ICC = 0.60). With regard to the provocative tests, agreement was best when apprehension was used as the criterion for a positive test and was better for the relocation (ICC = 0.71) and release tests (ICC = 0.63) than for the apprehension (ICC = 0.47) and augmentations tests (ICC = 0.48). Reliability was poor (ICC < 0.31) when pain was used as the criterion for a positive test. These results indicate that the load and-shift, sulcus, and provocative tests (apprehension, augmentation, relocation, and release) are reliable clinical tests for instability in symptomatic patients when care is taken with respect to arm positioning and if apprehension is used as the criterion for a positive provocative test. PMID- 14735069 TI - Shoulder electromyography in multidirectional instability. AB - We studied shoulder muscle activity in multidirectional instability (MDI) and multidirectional laxity (MDL) of the shoulder, our hypothesis being that altered muscle activity plays a role in their pathogenesis. Six muscles (supraspinatus, infraspinatus, subscapularis, anterior deltoid, middle deltoid, and posterior deltoid) were investigated by use of intramuscular dual fine-wire electrodes in 7 normal shoulders, 5 MDL shoulders, and 6 MDI shoulders. Each subject performed 5 types of exercise (rotation in neutral, 45 degrees of abduction, 90 degrees of abduction, flexion/extension, and abduction/adduction) on an isokinetic muscle dynamometer at two rates, 90 degrees /s and 180 degrees /s. After filtering, rectification, and smoothing, the electromyography signal was normalized by using the peak voltage of the movement cycle. In subjects with MDI, compared with normal subjects, activity patterns of the anterior deltoid were different during rotation in neutral and 90 degrees of abduction, whereas those of the middle and posterior deltoid were different during rotation in 90 degrees of abduction. In subjects with MDL, the posterior deltoid showed increased activity compared with normal subjects during adduction. Activity patterns of the supraspinatus, infraspinatus, and subscapularis appeared similar in both groups. Dual fine-wire electromyography offers insight into the complex role of shoulder girdle muscle function in normal movement and in instability. Altered patterns of shoulder girdle muscle activity and imbalances in muscle forces support the theory that impaired coordination of shoulder girdle muscle activity and inefficiency of the dynamic stabilizers of the glenohumeral joint are involved in the etiology of MDI. Interestingly, the abnormalities are in the deltoid rather than the muscles of the rotator cuff. PMID- 14735070 TI - Traumatic recurrent anterior dislocation of the shoulder: two- to four-year follow-up of an anatomic open procedure. AB - The purpose of this study was to evaluate the results of an anatomic open stabilization procedure. Twenty-three consecutive patients with traumatic, recurrent, anterior glenohumeral instability were managed with a modified open procedure. All had a Bankart lesion. A standardized, true anatomic capsulolabral reconstruction was performed with suture anchors. Two patients were lost to follow-up, and twenty-one were evaluated after 36 months [range, 24-49 months] by an independent observer. Shoulder stability and function were the principal outcome measures. None of the patients had subsequent dislocations. All returned to full working capacity, and 19 reached their previous level of sport activities. The mean loss in active abduction and flexion was 1 degrees, in internal rotation, one vertebral level, and in external rotation, 7 degrees (arm at 90 degrees of abduction). Two patients had a positive anterior apprehension test. No sign of shoulder osteoarthritis was observed. The open anatomic capsulolabral reconstruction provides excellent results and allows the restoration of stability with good function. PMID- 14735071 TI - The sublabral foramen: does it affect stress distribution on the anterior glenoid? AB - Even though it is believed that a sublabral foramen (SF) requires no treatment, no objective data are available to establish whether this condition bears a relationship to anterior-inferior glenohumeral instability. Therefore, the influence on glenoid subchondral bone mineralization of an isolated SF was investigated, because the individual distribution of subchondral bone mineralization may be used as an indirect parameter for long-term stress distribution of joints. Two age- and side-matched groups of healthy glenohumeral specimens with SF (n = 10, aged 37-85 years) and without SF (n = 10, aged 36-86 years) were examined by computed tomography osteoabsorptiometry. As variables for comparison, the anterior and posterior density maxima on the glenoid were measured in a standardized manner. No shift of the anterior density maximum [p(x1) = 0.353/p(y1) = 0.739] was found between both groups, which is in contrast to anterior glenohumeral instability. This indicates a long-term stress distribution in SF shoulders comparable to that in non-SF shoulders. The data suggest that an isolated SF is probably not disproportionately related to glenohumeral instability and support the general assumption that surgical treatment of SF is not required. PMID- 14735072 TI - Glenohumeral translations are increased after a type II superior labrum anterior posterior lesion: a cadaveric study of severity of passive stabilizer injury. AB - The effects of simulated type II superior labrum anterior-posterior (SLAP) lesions were studied to determine whether the severity of the lesion affected glenohumeral joint translations. A robotic/universal force-moment sensor testing system was used to simulate load-and-shift tests by applying an anterior or posterior load of 50 N to each shoulder. The apprehension test for anterior instability was simulated by applying an anterior load of 50 N with an external rotation torque of 3 Nm at 30 degrees and 60 degrees of abduction. This loading protocol was repeated after creating two type II SLAP lesions of different severity. In the first the superior labrum and the biceps anchor were elevated subperiosteally from the glenoid bone (SLAP-II-1), and in the second the biceps anchor was completely detached (SLAP-II-2). Statistical analysis was performed with a 2-factor repeated-measures analysis of variance followed by multiple contrasts, and the significance level was set at P <.05. At 30 degrees of abduction, anterior translation of the vented joint from anterior loading was 18.5 +/- 8.5 mm. It was significantly increased (26.2 +/- 6.5 mm, P =.03), after the SLAP-II-2 lesion and compared with the SLAP-II-1 lesion (25.0 +/- 6.8 mm, P =.03). Increases in anterior translations at 60 degrees of abduction were not significantly differ in comparison to the two SLAP lesions. Inferior translation also resulted from anterior loading. At 30 degrees of abduction in the vented joint, it was 3.8 +/- 4.0 mm and was significantly increased (8.5 +/- 5.4 mm, P =.05) after the SLAP-II-2 lesion, no different than that after the SLAP-II-1 lesion (7.8 +/- 4.9 mm). No significant increases in anterior translation occurred in response to the combined loading condition between the two SLAP lesions. Glenohumeral translation was increased, regardless of severity, after simulation of type II SLAP lesions. During stabilizing surgical interventions, passive stabilizers that are injured in the type II SLAP lesion should be considered as well as dynamic activity in the tendon of the long head of the biceps brachii. PMID- 14735073 TI - Contribution to the study of the pathogenesis of type II superior labrum anterior posterior lesions: a cadaveric model of a fall on the outstretched hand. AB - With a cadaveric model, we studied the effects on the superior labrum-biceps tendon complex of a fall on the outstretched hand to assess one of the supposed mechanisms of creation of superior labrum anterior-posterior (SLAP) lesions. Ten shoulders were used to simulate either a forward or a backward fall with a custom shoulder-testing apparatus capable of simulating muscle forces, attached to a servohydraulic testing machine (INSTRON 8500+). Impaction of the humeral head on the glenoid cavity was applied in 0.1 seconds. The presence or absence of a SLAP lesion was determined. For the 5 shoulders used to simulate a forward fall, 5 type II SLAP lesions were found; for the 5 shoulders used to simulate a backward fall, only 2 type II SLAP lesions were observed. The role of shearing forces seems to be a major factor in the pathogenesis of these lesions, in association with predisposing anatomic factors. PMID- 14735074 TI - Mechanical properties of regenerated coracoacromial ligament after subacromial decompression. AB - Recent publications suggest that the coracoacromial ligament regenerates after it has been partially excised during subacromial decompression or acromioplasty. This observation may aid the understanding of the successes and failures of this very commonly performed surgical procedure. This study determines the mechanical properties of the apparently regenerated ligament. Eight regenerated coracoacromial ligaments were excised during revision surgery after subacromial decompression and were taken for mechanical testing. It appears that the ligament does have the ability to re-form relatively quickly after subacromial decompression or acromioplasty but takes time to regain strength. The results indicate that the ligament may possibly regain normal mechanical properties after regeneration times in excess of 3 years. PMID- 14735075 TI - Regional antibiotic prophylaxis in elbow surgery. AB - In this study we describe a technique for the delivery of regional antibiotic prophylaxis in patients undergoing elbow surgery and compare tissue antibiotic concentrations achieved by this technique with those achieved by standard systemic intravenous prophylaxis. We collected bone and fat samples from patients undergoing elective elbow surgery who had received regional antibiotic prophylaxis and measured the tissue antibiotic concentration. For comparison, we measured the antibiotic concentration in bone and fat samples taken from patients undergoing elective shoulder surgery who had received systemic prophylaxis. Mean tissue antibiotic concentrations were significantly higher in the regional antibiotic group (bone, 1484 microg/g vs 35.8 microg/g; fat, 1422.7 microg/g vs 10.7 microg/g; P <.05). No adverse effects were encountered with regional antibiotic delivery. The delivery of regional antibiotic prophylaxis in elbow surgery achieves higher tissue antibiotic concentrations than those achieved with standard systemic delivery. This technique may help reduce the risk of perioperative infection in elbow surgery. PMID- 14735076 TI - Ulnar nerve palsy at the elbow after surgical treatment for fractures of the olecranon. AB - Despite close proximity of the fracture site to the nerve, ulnar nerve palsy after surgery for fracture of the olecranon is uncommon. We examined 18 cases of fracture of the olecranon treated surgically retrospectively to see if there were any characteristics common to 4 cases of ulnar nerve palsy. Three cases of palsy with a comminuted fracture had fair or poor reduction as revealed by both the lateral and anteroposterior radiographs taken at the time of surgery. The other case, with little displacement of the fragment, had osteoarthritic changes at the medial side of the elbow. The mechanism of development of ulnar nerve palsy is considered to be multifactorial. However, it should be noted that evaluation of reduction of the fracture, not only on a lateral radiograph but also on an anteroposterior radiograph at the time of surgery, is important to prevent the ulnar nerve from being jeopardized. PMID- 14735077 TI - Posterior olecranon resection and ulnar collateral ligament strain. AB - Valgus extension overload is well described in the throwing athlete. The surgical treatment involves posterior olecranon osteophyte resection. It is not known how much of the posterior olecranon can be resected before increased stress is placed on the ulnar collateral ligament. Ten cadaveric arms underwent posterior olecranon resection in 4-mm increments. After each bone resection, three different valgus loads were applied to the elbow at 70 degrees and 90 degrees of flexion. Ulnar collateral ligament strain significantly increased with each load. Significantly more strain occurred on the ulnar collateral ligament at 90 degrees compared with 70 degrees of elbow flexion. This difference may be the result of the relative importance of the ulnar collateral ligament and the posterior olecranon at different positions of elbow flexion. The strain on the ulnar collateral ligament was slightly higher after the 8-mm and 12-mm cuts but not statistically significant. This study suggests that at moderate quasistatic valgus loads, ulnar collateral ligament strain is not significantly increased with posterior olecranon resection. PMID- 14735078 TI - The Orthopaedic Research Institute-Tennis Elbow Testing System: A modified chair pick-up test-interrater and intrarater reliability testing and validity for monitoring lateral epicondylosis. AB - Lateral epicondylosis is a degenerative overuse tendinopathy involving the extensor tendons of the forearm, predominantly the extensor carpi radialis brevis, in the region of the lateral epicondyle of the elbow. The Orthopaedic Research Institute-Tennis Elbow Testing System (ORI-TETS) is designed to record objective measurements of force generated with a simulated chair pick-up test. Interrater reliability of the ORI-TETS was excellent, with high intraclass correlation coefficients (ICCs) for right arm mean peak force of 0.93, left arm mean peak force of 0.84, right arm mean total force of 0.93, and left arm mean total force of 0.86. The ORI-TETS also demonstrated excellent intrarater reliability, with ICCs ranging from 0.9 to 0.97. The relative technical error of the ORI-TETS for all measurements ranged from 5.8% to 7.2%. Testing patients with lateral epicondylosis (N = 16) and comparing analog pain scores with ORI-TETS testing demonstrated a strong negative relationship between the two parameters (Spearman rho, -0.87 to -1.0). Thus, the ORI-TETS is a reliable and reproducible testing system for the forearm extensors. The testing system is inexpensive, takes 5 minutes to perform, and demonstrates good predictive value for objectively assessing patients with lateral epicondylosis. This system could be used for routine clinical monitoring of patients with lateral epicondylosis. PMID- 14735079 TI - Radial head prosthesis with a mobile head. AB - Radial head replacement is a useful method in the treatment of comminuted radial head fractures. Because such injuries are a common clinical problem and many complications have been reported after radial head replacements, a new radial head prosthesis is proposed. This new design, based on the shape-dimensional identification of the radial head, consists of two parts. The head, made of ultra high molecular weight polyethylene, is articulated with a Vitallium stem. A series of functional quality and strength tests were conducted on this new prosthetic design. The implant was also examined via the finite element method. General preclinical investigations of clinical cases show that this prosthesis is a very promising design. PMID- 14735080 TI - Subchondral insufficiency fracture of the humeral head and glenoid resulting in rapidly destructive arthrosis: a case report. PMID- 14735081 TI - Extra-abdominal desmoid around the shoulder joint: one of the differential diagnoses of frozen shoulder. PMID- 14735082 TI - Snapping scapula syndrome due to malunion of rib fractures: a case report. PMID- 14735083 TI - Foreign-body reaction after reconstruction of complete acromioclavicular dislocation using PDS augmentation. PMID- 14735084 TI - Neglected elbow dislocation in a young man: treatment by open reduction and elbow fixator. PMID- 14735085 TI - Tuberculous bursitis of the subacromial bursa. PMID- 14735086 TI - Bilateral shoulder bursal osteochondromatosis associated with complete rotator cuff tear. PMID- 14735087 TI - Acromial impression fracture of the greater tuberosity with rotator cuff avulsion due to hyperabduction injury of the shoulder. PMID- 14735088 TI - Snapping scapula caused by abnormal angulation of the superior angle of the scapula. PMID- 14735089 TI - Shoulder function after subtotal excision of the deltoid muscle and of the deltoid and infraspinatus muscles in extra-abdominal desmoid tumor: a long-term follow-up in two cases. PMID- 14735090 TI - Displaced scapular neck fracture: a case report. PMID- 14735093 TI - Boo! Don't let mapping scare you. PMID- 14735092 TI - Not all scars are ugly. PMID- 14735094 TI - Priority traffic. PMID- 14735095 TI - A regular or irregular irregularity? PMID- 14735097 TI - Congenital heart defects in children. Part 1 of a 2-part series: anatomical anomalies & their resultant pathophysiology. PMID- 14735098 TI - Flash pulmonary edema: early recognition & prompt prehospital therapy improve patient outcomes. PMID- 14735106 TI - Memphis just says 'No!' to ambulance diversion. PMID- 14735107 TI - PharmGKB: the pharmacogenetics and pharmacogenomics knowledge base. PMID- 14735109 TI - Re: KC Wilhelmsen, M Schuckit et al (2003): the search for genes related to a low level response to alcohol determined by alcohol challenges. PMID- 14735110 TI - Promoting comparative molecular studies in environmental health research: an overview of the comparative toxicogenomics database (CTD). PMID- 14735111 TI - Multilocus effects of the renin-angiotensin-aldosterone system genes on blood pressure response to a thiazide diuretic. AB - BACKGROUND: The renin-angiotensin-aldosterone (RAA) system regulates blood pressure (BP) levels and influences responses to antihypertensive medications. Variation in RAA system genes has been reported to influence interindividual differences in BP levels and the occurrence of hypertension (HTN). METHODS: We evaluated the relationship between variation in genes of the RAA system and interindividual differences in BP response to a thiazide diuretic. Analyses were carried out in a race- and gender-specific manner in 255 unrelated hypertensive African-Americans (125 men and 130 women) and 246 unrelated hypertensive non Hispanic Whites (133 men and 113 women). RESULTS: The angiotensin II receptor (AT(1)R) A1166C and angiotensinogen G-6A polymorphisms had a significant effect on systolic BP response to the diuretic in African-American women. Multilocus analyses indicated that the effects of these genes combined additively to influence response. Results of a permutation test to adjust for multiple comparisons and the possible nonindependence among genotypes remained significant at the P=0.003 level. CONCLUSIONS: Among African-American women, particular gene variations in the RAA system have additive effects on BP response to a thiazide diuretic. PMID- 14735112 TI - Insights into the ageing mind: a view from cognitive neuroscience. PMID- 14735113 TI - Homeostatic plasticity in the developing nervous system. PMID- 14735114 TI - Pathways of the past: the imprint of memory. PMID- 14735115 TI - Neocortical cell classes are flexible entities. PMID- 14735116 TI - Presynaptic ionotropic receptors and control of transmitter release. PMID- 14735117 TI - Regeneration beyond the glial scar. PMID- 14735118 TI - Myelin inhibitors: does NO mean GO? PMID- 14735120 TI - The complex interplay among factors that influence allelic association. PMID- 14735121 TI - Network biology: understanding the cell's functional organization. PMID- 14735122 TI - Genes and speciation. PMID- 14735123 TI - Endosymbiotic gene transfer: organelle genomes forge eukaryotic chromosomes. PMID- 14735124 TI - Using human neural stem cells to model neurological disease. PMID- 14735125 TI - National traditions and the emergence of genetics: the French example. PMID- 14735126 TI - Increased anticholinergic challenge-induced memory impairment associated with the APOE-epsilon4 allele in the elderly: a controlled pilot study. AB - The degree to which elderly adults experience cognitive impairments from centrally acting anticholinergic drugs is variable, but the cause of this variability is unknown. The present study examined the epsilon4 allele as a possible modulator of the effects of trihexyphenidyl hydrochloride (Artane( trade mark )), an anticholinergic drug, on memory functioning. Of the 24 cognitively intact, elderly participants (age range 62-76), 12 who possessed the epsilon4 allele, participated in a double-blind, randomized, placebo-controlled, crossover, three-way study. All participants were tested after receiving a single oral dose of trihexyphenidyl (1 or 2 mg) or placebo, with a 7-day washout period between sessions. Memory and psychomotor tests were administered at baseline, and at 1, 2.5, and 5 h post-treatment. Results showed that participants with the epsilon4 allele demonstrated significant impairments in delayed recall after both 1 and 2 mg doses of trihexyphenidyl while the non-epsilon4 group did not. Additionally, while acute administration of the 2 mg dose significantly impaired total recall in both epsilon4 and non-epsilon4 carriers, the epsilon4 carriers showed a more persistent impairment. These findings held when participants with the epsilon2 allele were excluded from the analyses. The epsilon4 groups did not differ with respect to psychomotor performance or plasma drug levels. These results provide evidence suggesting that the epsilon4 allele plays a significant role in increasing cognitive sensitivity to trihexyphenidyl and that a temporal component of memory consolidation may be especially vulnerable. A larger study is warranted to confirm these preliminary findings. PMID- 14735127 TI - Testing early-onset chronic atypical depression subtype. PMID- 14735128 TI - Effects of chronic olanzapine and haloperidol differ on the mouse N1 auditory evoked potential. AB - Auditory evoked potentials have been used in a variety of animal models to assess information-processing impairments in schizophrenia. Previous mouse models have primarily employed a paired click paradigm to assess the transient measures of auditory gating. The current study uses stimulus trains at varied interstimulus intervals (ISI) between 0.25 and 8 s in mice to assess the effects of chronic olanzapine and haloperidol on auditory processing. Data indicate that olanzapine increases the amplitude of the N40, P80, and P20/N40 components of the auditory evoked potential, whereas haloperidol had no such effect. The ISI paradigm also allowed for an evaluation of several components of the mouse evoked potential to assess those that display response properties similar to the human P50 and N100. Data suggest that the mouse N40 displays an ISI response relationship that shares characteristics with the human N100, whereas the P20 appears more consistent with the human P50 across the ISI range evaluated in this task. This study suggests that olanzapine may help improve N100 impairments seen in schizophrenia, while haloperidol does not. PMID- 14735129 TI - Seizure expression during electroconvulsive therapy: relationships with clinical outcome and cognitive side effects. AB - Since electroconvulsive therapy (ECT) can result in generalized seizures that lack efficacy, physiological markers of treatment adequacy are needed. Specific electroencephalographic (EEG) features differentiate seizures produced with barely suprathreshold right unilateral (RUL) ECT, an ineffective treatment, from effective forms of ECT. This study determined whether EEG features are sensitive to treatment condition using a broad dosing range for RUL ECT, as well as predictive of clinical and cognitive outcomes. Quantitative EEG measures and observer ratings were compared in predictive power. From a larger study, 54 in patients with major depression were randomized to low (1.5 x seizure threshold (ST)), moderate (2.5 x ST), or high-dose (6 x ST) RUL ECT, or high-dose (2.5 x ST) bilateral (BL) ECT. High dosage RUL and BL ECT were comparable in efficacy, and superior to low and moderate dosage RUL ECT. In the slow frequency bands (delta), BL ECT resulted in greater ictal power, ictal coherence, and postictal suppression than each RUL ECT condition, but the EEG measures failed to discriminate the RUL ECT groups. EEG measures were modestly associated with clinical outcome, with greater ictal power, delta coherence, and postictal suppression positive predictors. None of the EEG measures were associated with cognitive outcomes. Inability to distinguish forms of RUL ECT differing markedly in dosage and efficacy suggests that EEG measures have limited potential as markers of treatment adequacy. Rather than assaying treatment adequacy, the EEG features associated with efficacy may reflect individual differences in the strength of inhibitory processes that terminate the seizure, and can help isolate the biological variability that predisposes to positive or negative clinical response to ECT. PMID- 14735130 TI - Regulation of the human corticotropin-releasing-hormone gene promoter activity by antidepressant drugs in Neuro-2A and AtT-20 cells. AB - Major depression is frequently associated with hyperactivity of the hypothalamic pituitary-adrenal (HPA) axis. Clinically effective therapy with antidepressant drugs normalizes the disturbed activity of HPA axis, in part, by decreasing corticotropin-releasing hormone (CRH) synthesis, but the mechanism of this action is poorly recognized. In order to find out whether antidepressants directly affect CRH gene promoter activity, we studied their effect on undifferentiated and differentiated Neuro-2A cells, and for comparison the effect of the selected antidepressants on AtT-20 cells was also determined. The cells were stably transfected with a human CRH promoter fragment (-663 to +124 bp) linked to the chloramphenicol acetyltransferase (CAT) reporter gene. The regulation of CRH gene promoter activity is similar in Neuro-2A cells, both intact and differentiated, and in AtT-20 cell line, and cAMP/PKA-dependent pathway plays an important role in the stimulation of CRH gene. It was found that imipramine, amitryptyline, desipramine, fluoxetine, and mianserin, present in the culture medium for 5 days, in a concentration-dependent manner inhibited basal hCRH gene promoter activity in undifferentiated Neuro-2A cells, while other drugs under study (citalopram, tianeptine, moclobemide, venlafaxine, reboxetine, mirtazapine, and milnacipram) were inactive. In the differentiated cells, all examined antidepressants, except moclobemide (no effect) and tianeptine (increase), inhibited hCRH gene transcription. Moreover, in differentiated cells, the drugs acted stronger and were effective at lower concentrations. Forskolin-induced CAT activity was attenuated by imipramine and fluoxetine and to a lesser degree by amitriptyline and desipramine in differentiated cells, whereas other drugs were inactive. Moreover, imipramine and fluoxetine, but not tianeptine, showed moderate inhibitory effect on CRH gene promoter activity also in AtT-20 cell line, commonly used in CRH gene regulation studies. These results indicate that neuron like differentiated Neuro-2A cells are a better model than pituitary and intact neuroblastoma to investigate the mechanism of psychotropic drug action. Inhibition of CRH gene promoter activity by antidepressant drugs may be a molecular mechanism by which these drugs inhibit the activity of HPA axis. PMID- 14735131 TI - Histamine h3 receptor antagonists potentiate methamphetamine self-administration and methamphetamine-induced accumbal dopamine release. AB - Methamphetamine administration increases brain levels of histamine and neuronal histamine attenuates several of methamphetamine's behavioral effects. The role of different subtypes of histamine receptors in this negative feedback, however, remains unclear. There is some evidence on possible involvement of histamine H3 receptors in these actions of methamphetamine. The aim of the present study was to evaluate the effects of two histamine H3 receptor antagonists, clobenpropit and thioperamide, on rewarding and neurochemical effects of methamphetamine utilizing three in vivo methodologies, drug self-administration, drug discrimination, and microdialysis in Sprague-Dawley rats. In rats self administering methamphetamine intravenously under a fixed-ratio schedule, presession treatment with thioperamide (1.0-3.0 mg/kg, subcutaneous, s.c.) or clobenpropit (1.0-3.0 mg/kg, s.c.) potentiated the reinforcing effects of methamphetamine, as indicated by a dose-dependent increase in responding for a low 0.03 mg/kg dose of methamphetamine, that by itself failed to maintain responding above saline substitution levels, and a decrease in responding for a higher 0.06 mg/kg training dose of methamphetamine. In contrast, neither thioperamide nor clobenpropit treatment increased responding during saline substitution. In other rats trained to discriminate intraperitoneal (i.p.) injection of 1.0 mg/kg methamphetamine from i.p. injection of saline, both thioperamide and clobenpropit (0.3-3.0 mg/kg, s.c.) dose dependently increased methamphetamine-appropriate responding when administered with a low 0.3 mg/kg i.p. dose of methamphetamine, which by itself produced predominantly saline appropriate responding. However, thioperamide and clobenpropit produced only saline-appropriate responding when administered with saline vehicle. Finally, thioperamide and clobenpropit potentiated methamphetamine-induced elevations in extracellular dopamine levels in the shell of the nucleus accumbens, but did not increase brain dopamine levels when given alone. These findings point to histamine H3 receptors as a new and important receptor system modulating the reinforcing, subjective, and neurochemical actions of methamphetamine. PMID- 14735132 TI - mGluR5 antagonist MPEP reduces ethanol-seeking and relapse behavior. AB - The glutamatergic system plays an important role in mediating neurobehavioral effects of ethanol. Metabotropic glutamate receptors subtype 5 (mGluR5) are modulators of glutamatergic neurotransmission and are abundant in brain regions known to be involved in ethanol self-administration. Here, we studied the effects of 2-methyl-6-(phenylethynyl)-pyridine (MPEP), a highly potent, noncompetitive mGlu5 receptor antagonist, on voluntary ethanol consumption and relapse behavior. For this purpose, we used two models for the measurement of relapse behavior: (i) reinstatement of ethanol-seeking behavior by drug-associated cues and (ii) the alcohol deprivation effect in long-term ethanol-consuming rats. In the first set of experiments, rats were trained to lever press for ethanol in the presence of a distinct set of cues. After extinction, the animals were exposed to the respective cues that initiated reinstatement of responding. A response-contingent ethanol prime further enhanced responding compared to the conditioned cues alone. Under these conditions, MPEP (0, 1, 3, and 10 mg/kg) attenuated ethanol seeking significantly and in a dose-related manner. However, at the highest dose, MPEP also decreased the number of inactive lever responses. In the second set of experiments, rats with 1 year of ethanol experience and repeated deprivation phases were used. A subchronic treatment with MPEP (twice daily; 0, 3, and 10 mg/kg) resulted in a significant and dose-dependent reduction of the alcohol deprivation effect (ADE). Although the same MPEP treatment regimen decreased baseline drinking, this effect was not as pronounced as on the ADE. These results show in two commonly used models of relapse to ethanol that pharmacological targeting of mGlu5 receptors may be a promising approach for the treatment of alcoholism. PMID- 14735133 TI - BDNF serum concentrations in healthy volunteers are associated with depression related personality traits. AB - The issue of neurotrophins is recognized as a new lead in the quest for a deeper understanding of mood disorders. This hypothesis has emerged from experimental evidence suggesting that antidepressant drugs might work by a neuroprotective effect through the stimulation of the neurotrophin expression in distinct regions of the CNS. Endogenous levels of BDNF protein were measured in the serum samples of 118 healthy unrelated volunteers (64 male, 54 female, age: 42.1+/-13.0 years), and the NEO-FFI has been performed in all subjects. BDNF serum values amounted to 16.3+/-7.3 ng/ml. BDNF concentration correlated significantly with age (r=0.182, p=0.048), but showed no gender differences (male 16.1+/-7.2, female 16.5+/-7.4 ng/ml). A negative correlation between the BDNF serum concentration and the depression-related factor neuroticism (r=-0.212, p=0.022) has been found. Low BDNF levels in healthy humans with depressive personality traits might constitute a risk marker, reflecting a personality profilethat is linked to vulnerability to mood disorders. These results provide further support for the hypothesis that BDNF may be central to the development of depressive mood states. PMID- 14735134 TI - Identification of lithium-regulated genes in cultured lymphoblasts of lithium responsive subjects with bipolar disorder. AB - Lithium, a common drug for the treatment of bipolar disorder (BD), requires chronic administration to prevent recurrences of the illness. The necessity for long-term treatment suggests that changes in genes expression are involved in the mechanism of its action. We studied effects of lithium on gene expression in lymphoblasts from BD patients, all excellent responders to lithium prophylaxis. Gene expression was analyzed using cDNA arrays that included a total of 2400 cDNAs. We found that chronic lithium treatment at a therapeutically relevant concentration decreased the expression of seven genes in lymphoblasts from lithium responders. Five of these candidate lithium-regulated genes, including alpha1B-adrenoceptor (alpha1B-AR), acetylcholine receptor protein alpha chain precursor (ACHR), cAMP-dependent 3',5'-cyclic phosphodiesterase 4D (PDE4D), substance-P receptor (SPR), and ras-related protein RAB7, were verified by Northern blotting analysis in lithium responders. None of these genes were regulated by lithium in healthy control subjects. When we compared the expression of these five genes between bipolar subjects and healthy control subjects at baseline, prior to lithium administration, we found that alpha1B-AR gene expression was higher in bipolar subjects than in healthy control subjects. Our findings indicate that alpha1B-AR may play an important role in the mechanism of action of lithium treatment. PMID- 14735135 TI - Drug-induced decrease of protein kinase a activity reveals alteration in BDNF expression of bipolar affective disorder. AB - Bipolar affective disorder (BAD) is a severe disease whose molecular and cellular bases are not well known. The aim of the present study was to probe the cAMP signaling downstream targets by pharmacologically manipulating the protein kinase A (PKA) enzyme, along with the assessment of brain-derived neurotrophic factor (BDNF) expression in lymphoblasts. The time course of lymphoblast PKA activity (up to 72 h) revealed optimal activity at 24 h. Then, the enzyme activity and protein levels of PKA Calpha subunit and phopsho-cAMP responsive element binding (CREB) were assayed in lymphoblasts derived from 12 BAD and 12 control (CT) subjects and cultured for 24 h in the presence of cAMP analog drugs. The results indicated that basal PKA activity and PKA Calpha subunit immunolabeling are increased in cells from BAD compared with controls. Enzyme activity was increased by Sp-isomer in BAD and in CT's cells, without change in protein levels. In contrast, the Rp-isomer decreased enzyme activity and protein levels. In drug naive conditions, there was no change in BDNF expression of BAD cells compared with CT cells. Treatment with Sp-isomer induced increased BDNF in both groups, while treatment with Rp-isomer induced a significant decrease in BDNF expression of BAD compared with CT. The p-CREB changes followed changes in BDNF levels, with increased and decreased Sp-isomer and Rp-isomer treatment, respectively. Our results suggest that mood disorder is associated with PKA upregulation and this could mask alteration in BDNF expression, because slowing down of PKA signaling results in a decrease of BDNF expression. These findings, combined with previous reports, provide a new insight to explain pharmacological features in different diagnostic groups. PMID- 14735136 TI - Effect of gonadectomy and gonadal hormone replacement on cocaine self administration in female and male rats. AB - Both sex and gonadal steroid hormones may influence the abuse-related behavioral effects of cocaine under some conditions, but there is considerable inconsistency in the literature. In the present study, rats were trained under a fixed ratio (FR) 5 schedule of food presentation and were then allowed to self-administer cocaine (1.0 mg/kg/injection) until behavior stabilized. Subsequently, complete dose-effect functions for cocaine self-administration (0.032-3.2 mg/kg/injection) were determined in female and male rats before and after gonadectomy, and in gonadectomized female and male rats before and during chronic treatment with estradiol or testosterone, respectively. Sex, gonadectomy, and gonadal hormones did not alter the shape or position of dose-effect functions for cocaine self administration. These results suggest that sex, estrogen, and testosterone levels are not critical determinants of cocaine's reinforcing effects in rats under these conditions. This study differed from earlier studies in that complete dose effect functions for cocaine were determined. These findings suggest that the behavioral training history, the unit dose of cocaine, and the schedule of reinforcement are important variables in studies of sex and gonadal hormone effects on cocaine self-administration. PMID- 14735137 TI - Evolutionary genetics of lifespan and mortality rates in two populations of the seed beetle, Callosobruchus maculatus. AB - The age at which individuals die varies substantially within and between species, but we still have little understanding of why there is such variation in life expectancy. We examined sex-specific and genetic variation in adult lifespan and the shape of mortality curves both within and between two populations of the seed beetle, Callosobruchus maculatus, that differ in a suite of life history characters associated with adaptation to different host species. Mean adult lifespan and the shape of the logistic mortality curves differed substantially between males and females (males had lower initial mortality rates, but a faster increase in the rate of mortality with increasing age) and between populations (they differed in the rate of increase in mortality with age). Larger individuals lived longer than smaller individuals, both because they had lower initial mortality rates and a slower increase in the rate of mortality with increasing age. However, differences in body size were not adequate to explain the differences in mortality between the sexes or populations. Both lifespan and mortality rates were genetically variable within populations and genetic variance/covariance matrices for lifespan differed between the populations and sexes. This study thus demonstrated substantial genetic variation in lifespan and mortality rates within and between populations of C. maculatus. PMID- 14735138 TI - Extreme polymorphism in a Y-linked sexually selected trait. AB - Males of the livebearing fish, Poecilia parae, exhibit one of the most complex polymorphisms known to occur within populations, whereas females are monomorphic. We describe five distinct male colour morphs and an associated size dimorphism, and demonstrate through pedigree analysis that the locus or loci controlling the male colour polymorphism is linked to the Y-chromosome. Field surveys from 1999 to 2002 of nine populations in Guyana and Suriname, South America, indicate that some morphs are consistently abundant and others are rare, implying that the colour polymorphism has important fitness consequences. By rearing offspring of field-inseminated females, we showed that the common morph is also the most successful morph in terms of reproduction. However, dichotomous choice tests show that two rare morphs are preferred by females over the common morph. These results suggest that alternative male mating strategies, sperm competition, overt male-male competition, or other processes are overriding female preferences in these populations. Furthermore, Y-linkage of the colour polymorphism in P. parae supports the hypothesis that heterogametic sex chromosomes harbour sexually antagonistic traits beneficial to the heterogametic sex. PMID- 14735139 TI - Analysis of cell proliferation in Drosophila wing imaginal discs using mosaic clones. AB - Experimental data on spatial and temporal distributions of mosaic clones in Drosophila wing imaginal disc were analyzed. Long-lived proliferation centers (PR1, PR2, and PR3) and areas with decreased proliferation activity were found in the notum region of the disc. Simulation of the growth kinetics of mosaic patches demonstrated that the cell cycle in proliferation centers PR2 and PR3 was shorter than the average cycle in the disc and in the center PR1. A nonrandom clustering of rapidly dividing cells was observed in the PR2, but not in the other cases. The reason why the cell-cycle duration and the clustering of dividing cells may not coincide is discussed in terms of the recruitment of nondividing cells into the cell cycle. The simulation of the time course of the first and second moments of the size distribution of mosaic clones allowed the variance of cell-cycle progression rates to be determined and demonstrated that a model with a continuous cell-cycle rates gave a better fit to the data than the transition probability model of Smith and Martin. PMID- 14735140 TI - Neutral locus heterozygosity, inbreeding, and survival in Darwin's ground finches (Geospiza fortis and G. scandens). AB - Comprehensive long-term studies of isolated populations provide valuable comparative data that may be used to evaluate different methods for quantifying the relationship between genetic diversity and fitness. Here, we report on data collected from large and well-characterized cohorts of the two numerically dominant species of Darwin's finches on Isla Daphne Major, Galapagos, Ecuador - Geospiza fortis and G. scandens. Multilocus microsatellite (SSR) genetic diversity estimates (heterozygosity and d2) and pedigree-based estimates of the inbreeding coefficient (f) were compared to each other and to two fitness components: lifespan and recruitment. In the larger sample of G. fortis, heterozygosity (H) was correlated with both fitness components, but no relationship was detected in the smaller sample of G. scandens. Analyses of the inbreeding coefficient detected highly significant relationships between f and recruitment, but no relationship between f and overall lifespan. The d2 statistic showed no relationship to either fitness component. When the two SSR-based estimators were compared to f, d2 was correlated with f in G. fortis in the predicted direction, while in G. scandens the relationship was positive. Multilocus heterozygosity was correlated with f in G. fortis but not in the G. scandens sample. A pedigree simulation demonstrated that the variation in true autozygosity can be large among individuals with the same level of inbreeding. This observation may supplement the interpretation of patterns relevant to the local (locus-specific) and general (genome-wide) effects hypotheses, which have been proposed to explain the mechanism responsible for associations between genetic diversity and fitness. PMID- 14735141 TI - Field and experimental evidence of preferential selfing in the freshwater mollusc Lymnaea truncatula (Gastropoda, Pulmonata). AB - We have conducted a thorough study of the mating system of Lymnaea truncatula, the intermediate host of the liver fluke, using three approaches: (i) a population genetics study, (ii) controlled pairings in the laboratory and (iii) a progeny-array analysis. The population genetics study revealed high levels of inbreeding in the studied populations, with strong clues that the extensive heterozygote deficiencies observed are due to selfing. However, Wahlund effects may also arise due to recolonisations from different source populations after bottleneck events. A breeding experiment helped to disentangle the mating system and the Wahlund effects, and showed that high levels of selfing occurred in isolation and in controlled pairings. However, the progeny-array analysis performed after a high-density culturing of the snails suggests that substantial outcrossing may also occur. PMID- 14735142 TI - Comparison of the fine-scale genetic structure of three dipterocarp species. AB - We investigated the fine-scale genetic structure of three tropical-rainforest trees, Hopea dryobalanoides, Shorea parvifolia and S. acuminata (Dipterocarpaceae), in Peninsular Malaysia, all of which cooccurred within a 6-ha plot in Pasoh Forest Reserve. A significant genetic structure was found in H. dryobalanoides, weaker (but still significant) genetic structure in S. parvifolia and nonsignificant structure in S. acuminata. Seeds of all three species are wind dispersed, and their flowers are thought to be insect pollinated. The most obvious difference among these species is their height: S. parvifolia and S. acuminata are canopy species, whereas H. dryobalanoides is a subcanopy species. Clear differences were also found among these species in their range of seed dispersal, which depends on the height of the release point; so taller trees disperse their seed more extensively. The estimates of seed dispersal area were consistent with the degree of genetic structure found in the three species. Therefore, tree height probably had a strong influence on the fine-scale genetic structure of the three species. PMID- 14735143 TI - The HLA-DR2 haplotype is associated with an increased proliferative response to the immunodominant CD4(+) T-cell epitope in human interferon-beta. AB - Human CD4(+) T-cell epitopes were identified in interferon-beta (IFN-beta)-1b. A prominent peptide epitope region was found that induced a proliferative response in 16% of all donors tested. Responses corresponded to the presence of the HLA DR2 haplotype. Responsive donors expressing the HLA-DQ6 allele showed an increased level of proliferation to the epitope as compared to peptide-responsive HLA-DQ6 negative donors. A similar result was found for HLA-DR15-expressing donors. PBMC from donors expressing HLA-DR15 were more likely to proliferate in response to IFN-beta in a whole-protein in vitro assay than donors who did not carry this haplotype. It is striking that the common DQ6 allele HLA-DQB1(*)0602 is found in linkage disequilibrium with HLA-DRB1(*)1501, and this combination defines the HLA genotype associated with the development of multiple sclerosis. The HLA association between a response to IFN-beta and MS might explain the prevalence of neutralizing antibody development, and may underlie the etiology of the disease. PMID- 14735144 TI - Functional correlates of the interleukin-1 receptor antagonist gene polymorphism in the colonic mucosa in ulcerative colitis. AB - Association studies have identified the interleukin-1 receptor antagonist gene allele 2(IL-1RN*2) as a marker of susceptibility in ulcerative colitis (UC). This study investigated the significance of the IL-1RN genotype with respect to protein and mRNA expression in the colonic mucosa. Homogenates of rectal biopsies from 99 UC and 54 controls were assayed for cytokines IL-1ra, IL-1a and IL-1b using ELISA. IL1RN, IL1A and IL1B genotypes were determined using restriction enzyme analysis. The ability of the two IL1RN alleles to generate steady-state mRNA accumulation was assessed in the colonic mucosa of seven heterozygous patients. Stepwise linear regression demonstrated that IL-1RN genotype (P=0.001), diagnosis (P<0.0001) and treatment (P<0.03) were independent factors associated with the IL-1ra protein level whilst IL1RN genotype (P=0.005) and macroscopic inflammatory grade (P<0.0001) were associated with the IL-1ra/ total IL-1 ratio. The IL1RN*2 correlated with reduced IL-1ra and IL-1ra/IL-1 ratio with a gene dosage effect. In heterozygous UC patients the ratio of allele 1 mRNA / allele 2 steady state mRNA was always greater than 1 (range: 1.2-3.1) (P=0.018). The IL 1RN*2 is associated with reduced levels of IL-1ra protein and IL-1RN mRNA in the colonic mucosa, providing a biologically plausible explanation for the observed association of the allele with the disease. PMID- 14735145 TI - Mining microarray data to identify transcription factors expressed in naive resting but not activated T lymphocytes. AB - Transcriptional repressors controlling the expression of cytokine genes have been implicated in a variety of physiological and pathological phenomena. An unknown repressor that binds to the distal NFAT element of the interleukin-2 (IL-2) gene promoter in naive T-helper lymphocytes has been implicated in autoimmune phenomena and has emerged as a potentially important factor controlling the latency of HIV-1. The aim of this paper was the identification of this repressor. We resorted to public microarray databases looking for DNA-binding proteins that are present in naive resting T cells but are downregulated when the cells are activated. A Bayesian data mining statistical analysis uncovered 25 candidate factors. Of the 25, NFAT4 and the oncogene ets-2 bind to the common motif AAGGAG found in the HIV-1 LTR and IL-2 probes. Ets-2 binding site contains the three G's that have been shown to be important for binding of the unknown factor; hence, we considered it the likeliest candidate. Electrophoretic mobility shift assays confirmed cross-reactivity between the unknown repressor and anti-ets-2 antibodies, and cotransfection experiments demonstrated the direct involvement of Ets-2 in silencing the IL-2 promoter. Designing experiments for transcription factor analysis using microarrays and Bayesian statistical methodologies provides a novel way toward elucidation of gene control networks. PMID- 14735147 TI - Functional genetic polymorphisms in cytokines and metabolic genes as additional genetic markers for susceptibility to develop type 1 diabetes. AB - Genetic association with type 1 diabetes (T1D) has been established for two chromosomal regions: HLA DQ/DR (IDDM1) and INS VNTR (IDDM2). To identify additional genetic markers, we tested polymorphisms in regulatory regions of several cytokine and important metabolic genes. These polymorphisms exhibit functional consequences for expression and function. Functional genetic polymorphisms of proinflammatory (T-helper-1: IL-2, IL-12 and IFN-gamma), anti inflammatory (T-helper-2: IL-4, IL-6 and IL-10) and metabolic (IGF-I, VDR and INS) genes were determined in 206 Dutch simplex families with juvenile onset T1D and the results were analysed using the transmission disequilibrium test. Significantly increased transmission to T1D probands was observed for the loci IDDM1, IDDM2 and the vitamin D receptor. Although none of the other individual polymorphisms was associated with disease individually, the combination of T helper-2 and metabolic/growth alleles IL-10(*)R2, IL-4(*)C, VDR(*)C and IGF I(*)wt was found to be transmitted more frequently than expected (67%, P(c)=0.015). We conclude that additional genetic predisposition to T1D is defined by combinations of markers (eg Th2 and metabolic) rather than by a single marker. The consequences of the increased transmission of a low Th2 expressing genotypes together with a normal Th1 profile may result in a net proinflammatory cytokine expression pattern. PMID- 14735146 TI - Regulation of murine Tap1 and Lmp2 genes in macrophages by interferon gamma is mediated by STAT1 and IRF-1. AB - The genes of the transporter associated with antigen processing (Tap)-1, and the low molecular weight peptide (Lmp)-2, are crucial for class I major histocompatibility complex function and share a common bidirectional promoter. In murine bone marrow-derived macrophages, interferon gamma (IFN-gamma) induced Tap 1 and upregulated Lmp-2, which is constitutively expressed at low levels. The IFN gamma-induction was independent of early gene synthesis. The mRNA induced by IFN gamma was very stable. In macrophages from STAT1 knockout mice, IFN-gamma did not induce the expression of Tap-1 or Lmp-2. Several areas in the promoter can be controlled by IFN-gamma, such as proximal and distal GAS boxes in the direction of the Tap-1 gene, NFgammaB and IRF-1 boxes. By making deletions of the promoter, we found that only the proximal GAS and IRF-1 boxes are required for IFN-gamma induction of Tap-1 and Lmp-2. Experiments using nuclear extracts from macrophages treated for 30 min with IFN-gamma and gel shift analysis indicated that STAT1 binds to the GAS box. The nuclear extracts from macrophages treated for at least 2 h with IFN-gamma bound to the IRF-1 box. These results indicate that both STAT1 and IRF-1 are required for the IFN-gamma induction of Tap-1 and Lmp-2 genes. PMID- 14735148 TI - Toll-like receptor 4 polymorphism and severity of atopy in asthmatics. AB - Endotoxin exposure may have a protective effect against asthma and atopy. An Asp299Gly polymorphism in the Toll-like receptor 4 (TLR4) gene reduces responsiveness to endotoxin. This study determined the effect of TLR4 polymorphism on the risk and severity of asthma and atopy. In all, 336 UK Caucasian families with > or = 2 affected sibs (physician's diagnosis of asthma and current medication use) and 179 Caucasians without asthma or a family history of asthma were genotyped using ARMS-PCR. No association of the TLR4 polymorphism was found with the risk of developing asthma, either in parent-affected sibling trios, or in case-control analyses (P>0.05). In the first affected asthmatic siblings, the atopy severity score (based on size and number of positive skin prick tests and specific IgE) was higher in those with the Asp/Gly or Gly/Gly genotypes (mean 1.8, s.d. 1.1, n=39) compared to those with the Asp/Asp genotype (mean 1.2, s.d. 1.0, n=279) (P=0.003, t-test). No associations were found with total IgE, FEV(1) % predicted, slope of FEV(1) response to methacholine or asthma severity score (P>0.05). This study confirms the previously observed lack of association of TLR4 polymorphisms with asthma. In contrast, the findings suggest that genetically determined hyporesponsiveness to endotoxin may increase atopy severity. PMID- 14735149 TI - Evidence for a cluster of genes on chromosome 17q11-q21 controlling susceptibility to tuberculosis and leprosy in Brazilians. AB - The region of conserved synteny on mouse chromosome 11/human 17q11-q21 is known to carry a susceptibility gene(s) for intramacrophage pathogens. The region is rich in candidates including NOS2A, CCL2/MCP-1, CCL3/MIP-1alpha, CCL4/MIP-1beta, CCL5/RANTES, CCR7, STAT3 and STAT5A/5B. To examine the region in man, we studied 92 multicase tuberculosis (627 individuals) and 72 multicase leprosy (372 individuals) families from Brazil. Multipoint nonparametric analysis (ALLEGRO) using 16 microsatellites shows two peaks of linkage for leprosy at D17S250 (Z(lr) score 2.34; P=0.01) and D17S1795 (Z(lr) 2.67; P=0.004) and a single peak for tuberculosis at D17S250 (Z(lr) 2.04; P=0.02). Combined analysis shows significant linkage (peak Z(lr) 3.38) at D17S250, equivalent to an allele sharing LOD score 2.48 (P=0.0004). To determine whether one or multiple genes contribute, 49 informative single nucleotide polymorphisms were typed in candidate genes. Family based allelic association testing that was robust to family clustering demonstrated significant associations with tuberculosis susceptibility at four loci separated by intervals (NOS2A-8.4 Mb-CCL18-32.3 kb-CCL4-6.04 Mb-STAT5B) up to several Mb. Stepwise conditional logistic regression analysis using a case/pseudo-control data set showed that the four genes contributed separate main effects, consistent with a cluster of susceptibility genes across 17q11.2. PMID- 14735150 TI - An asthma-associated genetic variant of STAT6 predicts low burden of ascaris worm infestation. AB - Th-2 immune mechanisms are involved in the pathology of asthma and in the protective immune response to parasitic worms. Common upregulating genetic variants of Th-2 immune signalling are risk factors for asthma, and we tested whether they may confer a counteradvantage in protecting against parasitic worms. We examined the intensity of infection by the parasitic worm, Ascaris lumbricoides, by microsopic counting of ascaris eggs in the stool of 614 schoolchildren from an area of endemic ascaris infection in China. We investigated the relationship between the intensity of ascaris infection and common, asthma-associated genetic variants of Th-2 and Th-1 immune signalling. Ascaris egg counts per gram of stool (epg), mean 1068 epg, ranged from barely detectable (<240 epg) to heavy (approximately 9600 epg) in a skewed distribution. Logistic regression, after exploratory discriminant analysis, showed a major association between a common genetic variant of the 3'-UTR regulatory elements of the signal transducer and transactivating factor (STAT6) (P=0.0002) and egg counts, at the 77 th centile. Linear regression after log transformation of egg counts confirmed a highly significant association with this STAT6 variant (P=0.001). Thus, a common, asthma-associated, genetic variant of the pivotal transduction and transactivating factor for Th-2 immune signalling, STAT6, predicts increased resistance to ascaris worm infection. The evolution of enhanced resistance to parasitic worm infection, through human genetic variation in Th-2 immune signalling, may represent one origin for asthma. PMID- 14735151 TI - Genome-wide scans for leprosy and tuberculosis susceptibility genes in Brazilians. AB - Genome-wide scans were conducted for tuberculosis and leprosy per se in Brazil. At stage 1, 405 markers (10 cM map) were typed in 16 (178 individuals) tuberculosis and 21 (173 individuals) leprosy families. Nonparametric multipoint analysis detected 8 and 9 chromosomal regions respectively with provisional evidence (P<0.05) for linkage. At stage 2, 58 markers from positive regions were typed in a second set of 22 (176 individuals) tuberculosis families, with 22 additional markers typed in all families; 42 positive markers in 50 (192 individuals) new leprosy families, and 30 additional markers in all families. Three regions (10q26.13, 11q12.3, 20p12.1) retained suggestive evidence (peak LOD scores 1.31, 1.85, 1.78; P=0.007, 0.0018, 0.0021) for linkage to tuberculosis, 3 regions (6p21.32, 17q22, 20p13) to leprosy (HLA-DQA, 3.23, P=5.8 x 10(-5); D17S1868, 2.38, P=0.0005; D20S889, 1.51, P=0.004). The peak at D20S889 for leprosy is 3.5 Mb distal to that reported at D20S115 for leprosy in India. (151 words). PMID- 14735152 TI - Epistatic effects of genes encoding immunoglobulin GM allotypes and interleukin-6 on the production of autoantibodies to 60- and 65-kDa heat-shock proteins. AB - Immunoglobulin GM and KM genes have been associated with antibody responses to a variety of antigens. A promoter-region polymorphism of interleukin-6 (IL-6) gene (-174 G/C) has been shown to be associated with antibody responses to heat-shock proteins (hsp) 60 and hsp65. To examine the possible epistatic effects of these unlinked genetic systems on the autoimmune responses to hsp60 and hsp65, 176 healthy Caucasian subjects from Finland were genotyped for several allelic determinants of GM, KM, and IL-6 genes by PCR-RFLP methods. IgG antibodies to hsp60 and hsp65 were measured by an ELISA. Significant interactive effects of GM f,z and IL-6-174 genotypes were noted for both anti-hsp60 (P=0.002) and anti hsp65 (P=0.038) antibody levels. Since these autoantibodies have been implicated in susceptibility to coronary heart disease and carotid atherosclerosis, the associations reported here might be relevant to the etiology of these diseases. PMID- 14735153 TI - A vulnerability locus to multiple sclerosis maps to 7p15 in a region syntenic to an EAE locus in the rat. AB - Multiple sclerosis (MS) is a chronic immune-mediated demyelinating disease of the central nervous system. Evidence from family studies indicates a strong genetic component. Despite many studies of candidate genes, only an association with the HLA-DRB1*1501-DQB1*0602 haplotype has been generally detected, and HLA linkage established by transmission disequilibrium testing. A genome-wide scan revealed suggestive linkage of MS with markers on chromosome 7p15 in HLA-DR15-nonsharing British families, in a region syntenic to a locus predisposing to experimental autoimmune encephalomyelitis in the rat. We therefore tested the 7p15 region as a candidate region for genetic susceptibility to MS in 104 French families with at least two affected siblings. We found evidence suggestive of a predisposing locus in families in which only one affected sibling or none of them carry the HLA-DR15 allele. Comparison of the results of the British and French groups suggests that the region of interest can be narrowed to a 2.45-cM interval. PMID- 14735154 TI - Discrimination of suballeles present at the TNFd microsatellite locus using induced heteroduplex analysis. AB - Polymorphism at the TNFd locus has been implicated in a number of disease association studies. The TNFd locus consists of three regions of (GA)(n) repeats separated by an imperfect repeat of two guanine bases. TNFd alleles are genotyped by the number of repeats in the first (GA)(n) repeat region, and until now the second repeat region had been thought to be nonpolymorphic. We report the existence of suballeles present within the TNFd microsatellite locus, detected using induced heteroduplex generator (IHG) technology. These alleles cannot be detected using conventional typing strategies as they represent altered distribution of the (GA)(n) repeats or sequence variation within the repeat. The suballeles affect the frequencies of the conventional d3 and d4 alleles leading to significantly altered allele frequencies. Some studies have associated the d3 and d4 alleles with disease outcome. We re-analysed one such study cohort using IHG technology and demonstrated a high proportion of incorrectly assigned TNFd3 alleles. PMID- 14735155 TI - Association of the gene encoding the delta-subunit of the muscle acetylcholine receptor (CHRND) with acquired autoimmune myasthenia gravis. AB - The muscle acetylcholine receptor (AChR) is the main target self-antigen in acquired autoimmune myasthenia gravis (MG). Here, we investigated an association of MG with the CHRND gene encoding the delta-subunit of the AChR. Using a microsatellite repeat located in the second intron of the gene, we observed a preferential transmission of the allele 268 in 114 one-generation families with one myasthenic child (Pc=0.0154). This allele was also over-represented in a group of 350 unrelated nonthymoma MG patients (OR=1.78, P=0.038), but not in 84 thymoma patients, compared to 168 healthy controls. Moreover, among nonthymoma patients, those lacking serum anti-titin antibodies appeared to be best associated (OR=2.07, P=0.017). In contrast, there was no distortion in the transmission of a single-nucleotide substitution polymorphisms (SNPs) in the 3' untranslated region of CHRND nor in that of two SNPs located in the closely linked CHRNG gene, 4.5 kb telomeric to CHRND. The data warrant a detailed investigation of CHRND polymorphism in MG patients. PMID- 14735156 TI - RHD maternal-fetal genotype incompatibility and schizophrenia: extending the MFG test to include multiple siblings and birth order. AB - Rh incompatibility disease (ie Rh hemolytic disease of the fetus and newborn) has been implicated as a risk factor for schizophrenia. Here, we extend the maternal fetal genotype incompatibility (MFG) test used in an earlier case-parent trio study that found significant evidence for an increased risk of schizophrenia in RHD MFG-incompatible children. We modify the MFG test for case-parent trios to include any number of siblings. This modified test enables us to use more of the available data from the earlier study. The increased sample size not only gives us greater power to test for MFG incompatibility but it also enables us to model the impact of previous RHD MFG-incompatible pregnancies on the relative risk of RHD MFG incompatibility in later-born siblings. This modeling is important, because RHD MFG incompatibility is a proxy for Rh incompatibility disease, and the risk of Rh incompatibility disease increases with the number of previous RHD MFG-incompatible pregnancies. The best-fitting models are consistent with the hypothesized effect that previous incompatible pregnancies increase the risk of schizophrenia due to RHD MFG incompatibility. There was significant evidence that the relative risk of schizophrenia in the second- and later-born RHD MFG incompatible children is 1.7, consistent with earlier estimates. Our extension of the MFG test has general application to family-based studies of maternal-genotype and MFG interaction effects. PMID- 14735157 TI - The deleterious G15498A mutation in mitochondrial DNA-encoded cytochrome b may remain clinically silent in homoplasmic carriers. AB - We report on a patient with severe growth retardation and IgF1 deficiency, in which a mitochondrial abnormality was suspected. An isolated mitochondrial respiratory chain complex III deficiency was found in blood lymphocytes and skin fibroblasts. Sequence analysis of the cytochrome b, which is the only mitochondrial DNA-encoded subunit of complex III, revealed a homoplasmic G15498A mutation, resulting in the substitution of a highly conserved amino acid (glycine 251 into an aspartic acid). The mutation was found to be homoplasmic in all tissues examined from the mother and her brother (lymphocytes, fibroblasts, hair roots and buccal cells). Complex III deficiency was also demonstrated in these cells. Nevertheless, the mother and the brother were asymptomatic. This mutation had been considered as a cardiomyopathy-generating mutation in a previously reported case, and its pathogenicity has been demonstrated recently in yeast. However, it seems not to fulfil the classical criteria for pathogenicity of a mitochondrial DNA mutation, especially the heteroplasmic status, and to be clinically silent, albeit present, in nonaffected relatives. We suggest that other factors are contributing to the clinical variability expression of the G15498A mtDNA mutation. PMID- 14735158 TI - Surfactant protein C gene variation in the Finnish population - association with perinatal respiratory disease. AB - Surfactant protein C (SP-C) is a small hydrophobic protein component of alveolar surfactant, a lipid-protein complex lining the alveolar surface of the lung. Surfactant deficiency is the main cause of respiratory distress syndrome (RDS) in premature infants. RDS is a major risk factor of a chronic lung disease called bronchopulmonary dysplasia (BPD). The dominant mutations of the SP-C gene have recently been associated with interstitial lung diseases. However, the common genetic variation in the surfactant protein C gene has not been studied in detail. In the present study, the exonic variation of the SP-C gene in the Finnish population (n=472) was defined, and the association of the allelic variants with the susceptibility to RDS and BPD was examined. Conformation sensitive gel electrophoresis (CSGE) was used to determine the extent of exonic variation in the SP-C gene. Methods of genotyping were generated for three biallelic polymorphisms of the SP-C gene's exons 1, 4 and 5, which encode proSP C. The frequencies of these polymorphisms were evaluated in a study population consisting of 158 DNA samples from full-term infants. In addition, the linkage disequilibrium between the SP-C alleles was evaluated by haplotype analysis of parent-infant triplets. The role of SP-C gene variation in RDS and in BPD was evaluated in a high-risk population of 245 premature infants. According to the present results, the SP-C polymorphisms were associated with RDS and with very premature birth. The strength of allelic associations differed according to the gender of the premature infants. PMID- 14735159 TI - Is haplotype tagging the panacea to association mapping studies? PMID- 14735160 TI - Who supports the support workers? Cross-sectional survey of support workers' experience and views. AB - Support groups provide information and emotional support to families. Despite a recent growth in the number and size of these groups, there are no formal structures in place to provide support for the support worker. We performed a cross-sectional survey using a self-completion postal questionnaire, with the aim of identifying the structure, training needs and support given to workers. The participants were support workers from 112 United Kingdom-based organisations listed on the 'Contact a Family' website (www.cafamily.co.uk). We received 104 replies from 50/112 organisations (44%). Of these, 94/104 (90%) worked from home as volunteers. Two-thirds, 69/104, admitted times when they struggled to cope. A total of 43 (41%) admitted occasions of concern over the care given by a client to their affected relative. No group employed a professional to act in a clinical supervisory role. Our study suggests that support workers are highly committed to their role; these workers need support to ensure that they give appropriate advice under difficult circumstances. PMID- 14735161 TI - Serotonin transporter 5HTTLPR polymorphism and affective disorders: no evidence of association in a large European multicenter study. AB - The available data from preclinical and pharmacological studies on the role of the serotonin transporter (5-HTT) support the hypothesis that a dysfunction in brain serotonergic system activity contributes to the vulnerability to affective disorders (AD). 5-HTT is the major site of serotonin reuptake into the presynaptic neuron, and it has been shown that the polymorphic repeat polymorphism in the 5-HTT promotor region (5-HTTLPR) may affect gene transcription activity. 5-HTT maps to chromosome 17 at position 17q11.17-q12, and the 5-HTTLPR polymorphisms have been extensively investigated in AD with conflicting results. The present study tested the genetic contribution of the 5 HTTLPR polymorphism in a large European multicenter case-control sample, including 539 unipolar (UPAD), 572 bipolar patients (BPAD), and 821 controls (C). Our European collaboration has led to efforts to optimize a methodology that attenuates some of the major limitations of the case-control association approach. No association was found with primary psychiatric diagnosis (UPAD and BPAD) and with phenotypic traits (family history of AD, suicidal attempt, and presence of psychotic features). Our negative findings are not attributable to the lack of statistical power, and may contribute to clarify the role of 5-HTTLPR polymorphism in AD. PMID- 14735162 TI - Expansion to full mutation of a FMR1 intermediate allele over two generations. AB - Fragile X syndrome is due to an expanded CGG repeat in the 5' UTR of the FMR1 gene. According to repeat size, we distinguish four allele categories: normal (<40 CGG), intermediate (46-60 CGG), premutated (55-200 CGG) and full mutated (>200 CGG). However, the boundaries among these categories are unclear, making it difficult to classify unstable alleles and to estimate the risk of expansion. We report a family with a proband, carrying a methylated full mutation with an amplification of 1.2 kb. PCR analysis demonstrated two alleles of 29 and 61 CGGs in the mother. Sequencing of the 61 CGG allele showed no AGG interruptions. Both mother's sisters had two alleles of 31 and 44 CGGs, and the daughter of one of these had two alleles of 22 and 44 repeats, demonstrating stable transmission of the 44 CGG allele. The maternal grandfather was deceased, but haplotype reconstruction using markers DXS548 and FRAXAC1 demonstrated that he was carrier of the premutated allele. Furthermore, molecular analysis confirmed the same paternity with a probability of 99.79% for all the three sisters. According to these findings, it is likely that the maternal grandfather carried the 44 CGG allele, showing unstable transmission, given that it expanded first to 61 CGGs in one daughter, and then to full mutation in her child. Although we cannot exclude paternal mosaicism, it is likely that a rare event of progression from an intermediate to a premutated and on to a full mutated allele occurred in this family over two generations. PMID- 14735163 TI - Exclusion of an extracolonic disease modifier locus on chromosome 1p33-36 in a large Swiss familial adenomatous polyposis kindred. AB - Familial adenomatous polyposis (FAP), an autosomal dominantly inherited colorectal cancer predisposition syndrome, displays considerable inter- and intrafamilial phenotypic heterogeneity, which represents a major problem in genetic counselling of APC mutation carriers. The Min mouse model indicated a putative disease modifier locus on chromosome 4, which is syntenic to human chromosome 1p35-36. This finding was subsequently supported by parametric and nonparametric linkage analyses in FAP families, however, without identifying functional variants in candidate genes. Recently, germline mutations in the base excision repair gene MYH (1p33-34) have been described in patients with multiple adenomas, pointing to a possible role as disease modifier in FAP. Here, we present critical reassessment of one of the largest FAP kindreds published, which was previously used in linkage mapping of 1p35-36. In this family, all affected members harbour the same APC germline mutation (5945delA), but display marked phenotypic variability, in particular regarding the occurrence of extracolonic disease that segregates in several branches of the family tree. Using updated clinical information, additional mutation carriers and polymorphic markers, fine mapping of the critical region as well as mutation analysis of the MYH gene were performed. These investigations allowed us to significantly exclude (i) the 1p33 36 region as a modifier locus and (ii) MYH as a modifier gene for extracolonic disease in this FAP kindred. Our results do not eliminate 1p33-36 from suspicion in other families, but clearly indicate that in our family linkage analysis of further putative candidate regions is necessary to identify a disease modifier locus in FAP. PMID- 14735164 TI - Conferring specificity on the ubiquitous Raf/MEK signalling pathway. AB - The Raf-MEK-ERK signalling pathway controls fundamental cellular processes including proliferation, differentiation and survival. It remains enigmatic how this pathway can reliably convert a myriad of extracellular stimuli in specific biological responses. Recent results have shown that the Raf family isoforms A Raf, B-Raf and Raf-1 have different physiological functions. Here we review how Raf isozyme diversity contributes to the specification of functional diversity, in particular regarding the role of Raf isozymes in cancer. PMID- 14735165 TI - Negative regulation of ErbB family receptor tyrosine kinases. AB - Receptors of the EGF receptor or ErbB family of growth factor receptor tyrosine kinases are frequently overexpressed in a variety of solid tumours, and the aberrant activation of their tyrosine kinase activities is thought to contribute to tumour growth and progression. Much effort has been put into developing inhibitors of ErbB receptors, and both antibody and small-molecule approaches have exhibited clinical success. Recently, a number of endogenous negative regulatory proteins have been identified that suppress the signalling activity of ErbB receptors in cells. These include intracellular RING finger E3 ubiquitin ligases such as cbl and Nrdp1 that mediate ErbB receptor degradation, and may include a wide variety of secreted and transmembrane proteins that suppress receptor activation by growth factor ligands. It will be of interest to determine the extent to which tumour cells suppress these pathways to promote their progression, and whether restoration of endogenous receptor-negative regulatory pathways may be exploited for therapeutic benefit. PMID- 14735166 TI - Second primary breast cancer after Hodgkin's disease. AB - Although the potential carcinogenic risk of radiotherapy is well known, it has become clear that there is a particularly high risk of radiation-induced breast cancer in women treated for Hodgkin's disease at young ages. Thankfully, death from breast cancer in this population is uncommon, but it is important to understand factors contributing to the risk, including treatment parameters, and to develop a logical and efficient method for medical management of those at risk. In this minireview, we examine the evidence which should inform such a management policy. PMID- 14735167 TI - Modifiable risk factors for cancer. AB - Over 6 million people around the world die from cancer each year. Modifiable risk factors have been linked to a wide range of malignancies, including cancers of the oropharynx, oesophagus, larynx, lung, kidney, bladder, pancreas, skin, stomach, ovary, breast, cervix, uterus, prostate, and colon. Research indicates that over half of all cancers in developed countries could be prevented if we implemented population-wide measures to promote the following behaviours: reduce tobacco use, increase physical activity, control weight, improve diet, limit alcohol, utilise safer sex practices, get routine cancer screening tests, and avoid excess sun exposure. PMID- 14735168 TI - UFT/leucovorin and oxaliplatin alternated with UFT/leucovorin and irinotecan in metastatic colorectal cancer. AB - A total of 41 metastatic colorectal cancer (CRC) patients received tegafur/uracil (UFT)+leucovorin (LV)+oxaliplatin alternated with UFT/LV+irinotecan. The overall response rate was 58.5% (95% confidence interval, 42.2-73.3%), and the median progression-free survival was 8.8 months. There were no grade 4 toxicities; 12 patients (29%) experienced grade 3 diarrhoea. There were no cases of hand-foot syndrome. This alternating regimen seems to be effective and well tolerated in the first-line treatment of patients with metastatic CRC. PMID- 14735169 TI - Management of major depression in outpatients attending a cancer centre: a preliminary evaluation of a multicomponent cancer nurse-delivered intervention. AB - A novel nurse-delivered multicomponent intervention for major depressive disorder (MDD) in cancer outpatients was compared with usual care alone in a nonrandomised matched group design (n=30 per group). At the final 6-month outcome, 38.5% (95% CI, 5.4-57%) fewer patients in the intervention group still met the criteria for MDD. PMID- 14735170 TI - Major depression in outpatients attending a regional cancer centre: screening and unmet treatment needs. AB - A screening programme designed to identify cases of Major Depressive Disorder (MDD) in patients attending a Regional Cancer Centre outpatient department was established. It comprised two stages: (1) The Hospital Anxiety and Depression Scale (HADS) self-rating questionnaire administered by a touch-screen computer; (2) we interviewed patients with high scores on the HADS (15 or more total score) over the telephone using the depression section of the Structured Clinical Interview for DSMIV (SCID). A large consecutive sample (5613) of oncology clinic attenders was screened, and practical difficulties in the screening process were identified. The estimated prevalence of major depressive disorder (MDD) in the sample surveyed was approximately 8% (7.8%; 95% confidence intervals 6.9-8.5%). We assessed a consecutive series of 150 patients identified as having MDD to determine how many had received evidence-based treatment for MDD. Only half had discussed their low mood with their general practitioner, only one-third had been prescribed any antidepressant medication, and very few had taken a therapeutic dose for an adequate period. Very few had received psychological treatment or had been referred to mental health services. Most were receiving no potentially effective therapy. PMID- 14735171 TI - Communication and information-giving in high-risk breast cancer consultations: influence on patient outcomes. AB - This longitudinal study aimed to document (i) the information-giving and patient communication styles of clinical geneticists and genetic counsellors (consultants) in familial breast cancer clinics and (ii) assess the effect of these styles on women's knowledge, whether their expectations were met, satisfaction, risk perception and psychological status. A total of 158 women from high-risk breast cancer families completed self-report questionnaires at 2 weeks preconsultation and 4 weeks postconsultation. The consultations were audiotaped, transcribed and coded. Multivariate logistic regressions showed that discussing prophylactic mastectomy (P=0.00) and oophorectomy (P=0.01) led to women having significantly more expectations met; discussing genetic testing significantly decreased anxiety (P=0.03) and facilitating understanding significantly decreased depression (P=0.05). Receiving a summary letter of the consultation significantly lowered anxiety (P=0.01) and significantly increased the accuracy of perceived risk (P=0.02). Women whose consultant used more supportive communications experienced significantly more anxiety about breast cancer at the 4 weeks follow up (P=0.00). These women were not significantly more anxious before genetic counselling. In conclusion, this study found that consultants vary in the amount of information they give and the way they communicate; and this variation can result in better or worse psychosocial outcomes. Greater use of supportive and counselling communications appeared to increase anxiety about breast cancer. Identifying methods to assist consultants to address emotional issues effectively may be helpful. PMID- 14735172 TI - Patient-physician communication concerning participation in cancer chemotherapy trials. AB - Cancer patients demand a high level of involvement in decisions concerning treatment. Many patients are informed about experimental trials, and especially the first consultation may be crucial for the future communication and treatment process. Patients with nonresectable non-small-cell lung cancer or colorectal cancer informed about experimental chemotherapy completed a questionnaire on satisfaction with the communication process, general attitude towards experimental treatments, the substance of information, and personal contact with the physician following their first consultation in a medical oncology unit. Physicians completed a questionnaire on their perception of the patients' satisfaction. Among 68 physician-cancer patient pairs, 29 patients were informed on chemotherapy in randomised trials and 39 in nonrandomised studies. The general attitude towards experimental treatment was positive or very positive in 71% of patients. Information on the treatment was perceived as completely adequate in 93% of patients informed on randomised and in 67% informed on nonrandomised trials. Physicians underestimated the patients' satisfaction with the overall communication process, the personal contact, the patients' perceived sufficiency of the specific treatment information and their ability to decide on study entry. In conclusion, considerable differences were observed between patients informed about experimental chemotherapy in randomised and nonrandomised trials, both with respect to their perception of how adequate the information on the specific treatments were, and whether it was sufficient for decisions on study entry. This study type effect should be accounted for in future evaluations of communication and patient satisfaction. The data also support the fact that cancer patients have a desire for and ability to understand rather detailed and comprehensive treatment information. PMID- 14735173 TI - Randomised trial of a decision aid and its timing for women being tested for a BRCA1/2 mutation. AB - The aim of the study was to evaluate the impact of a decision aid (DA) and its timing in women being tested for a BRCA1/2 mutation. Women with and without a previous history of cancer were included after blood sampling for genetic testing. The DA consisted of a brochure and video providing information on screening and prophylactic surgery. To evaluate the impact of the DA, women were randomised to the DA group (n=184), receiving the DA 2 weeks after blood sampling, or to the control group (n=184). To evaluate the impact of timing, mutation carriers who had received the DA before the test result (n=47) were compared to mutation carriers who received the DA after the test result (n=42). Data were collected on well-being, treatment choice, decision and information related outcomes. The impact of the DA was measured 4 weeks after blood sampling. The impact of timing was measured 2 weeks after a positive test result. The DA had no impact on well-being. Regarding decision related outcomes, the DA group more frequently considered prophylactic surgery (P=0.02) corroborated with higher valuations (P=0.04). No differences were found for the other decision related outcomes. Regarding information related outcomes, the DA group felt better informed (P=0.00), was more satisfied with the information (P=0.00), and showed more accurate risk perceptions. Timing of the DA had no effect on any of the outcomes. No interactions were found between the DA and history of cancer. In conclusion, women being tested for a BRCA1/2 mutation benefit from the DA on information related outcomes. Because timing had no effect, the DA is considered useful either before or after the test result. PMID- 14735174 TI - Factors affecting pharmacokinetic variability of oral topotecan: a population analysis. AB - The aim of this study was to characterise the pharmacokinetics of the anticancer agent topotecan, and explore the influence of patient covariates and interoccasion variability on drug disposition. Data were obtained from 190 patients who received the drug as a 30-min infusion (N=72) or orally (N=118). The population model was built with the use of NONMEM to identify candidate covariates, and obtain models for clearance (CL) and volume of distribution. The final models were based on first-order absorption with lag-time (oral data), and a two-compartment model with linear elimination from the central compartment. The Cockcroft-Gault creatinine clearance (CrCl) and WHO performance status (PS) were the only significant covariates: CL=(12.8+2.1 x CrCl) x (1-0.12 x PS). For the volume of distribution, a correlation was found between body weight and the central volume (V1)=0.58 x body weight. Based on the structural models, a limited sampling strategy was developed with minor bias and good precision that can be applied a posteriori using timed samples obtained at 1.5, and 6 h after the administration of topotecan. In conclusion, a population pharmacokinetic model for topotecan has been developed that incorporates measures of renal function and PS to predict CL. In combination with drug monitoring, the limited sampling strategy allows individualised treatment for patients receiving oral topotecan. PMID- 14735175 TI - Phase I trial of concurrent chemoradiotherapy with docetaxel, cisplatin and 5 fluorouracil (TPF) in patients with locally advanced squamous cell carcinoma of the head and neck (SCCHN). AB - The aim of this study was to evaluate the efficacy and toxicity of a concurrent chemoradiotherapy using docetaxel, cisplatin and 5-fluorouracil (5-FU) (TPF) in patients with locally advanced squamous cell carcinoma of the head and neck (SCCHN). In total, 19 patients with previously untreated stage III-IV SCCHN were entered onto this trial. Patients received two cycles of chemotherapy. Cycles were repeated every 4 weeks. The starting doses (dose level 1) were docetaxel 60 mg m(-2), cisplatin 70 mg m(-2), and 5-day continuous infusion of 5-FU 600 mg m( 2) day(-1). Radiation was targeted to begin on the first day of chemotherapy, day 1. The total radiation dose to the primary tumour site and neck lymph nodes was between 63.0 and 74.0 Gy. At least three patients were examined at each dose level before advancing to the next level. The maximum-tolerated dose (MTD) of this regimen was docetaxel 60 mg m(-2), cisplatin 60 mg m(-2) and 5-FU 600 mg m( 2) day(-1). The main toxicities were mucositis (grade 3 and 4, 79%), leukocytopenia (grade 3 and 4, 53%), neutropenia (grade 3 and 4, 42%), anaemia (grade 3, 16%), liver dysfunction (grade 3, 11%) and renal dysfunction (grade 2, 11%). The overall response rate was 100%, including 84% complete responses (CRs). This concurrent chemoradiotherapy with TPF was safe and well tolerated. The high CR rate justifies further evaluation of this chemoradiotherapy modality in advanced SCCHN patients. PMID- 14735176 TI - Area under the curve of methotrexate and creatinine clearance are outcome determining factors in primary CNS lymphomas. AB - Although high-dose methotrexate (HD-MTX) is the most effective drug against primary CNS lymphomas (PCNSL), outcome-determining variables related to its administration schedule have not been defined. The impact on toxicity and outcome of the area under the curve (AUC(MTX)), dose intensity (DI(MTX)) and infusion rate (IR(MTX)) of MTX and plasmatic creatinine clearance (CL(crea)) was investigated in a retrospective series of 45 PCNSL patients treated with three different HD-MTX-based combinations. Anticonvulsants were administered in 31 pts (69%). Age >60 years, anticonvulsant therapy, slow IR(MTX) (1100 micromol hl(-1) were independently associated with a better survival. Slow CL(crea) and high AUC(MTX) are favourable outcome-determining factors in PCNSL, while slow CL(crea) is significantly related to higher toxicity. AUC(MTX) significantly correlates with age, anticonvulsant therapy, IR(MTX), and DI(MTX). These findings, which seem to support the choice of an MTX dose >/=3 gm(-2) in a 4-6-h infusion, every 3-4 weeks, deserve to be assessed prospectively in future trials. MTX dose adjustments in patients with fast CL(crea) should be investigated. PMID- 14735177 TI - A randomised phase II study of weekly paclitaxel or vinorelbine in combination with cisplatin against inoperable non-small-cell lung cancer previously untreated. AB - Phase II studies have suggested that weekly paclitaxel has a higher response rate and better toxicity profile than the conventional schedule of once every 3 or 4 weeks. Our aim was to evaluate the efficacy of weekly paclitaxel plus cisplatin (PC) vs vinorelbine plus cisplatin (VC) in chemonaive non-small-cell lung cancer (NSCLC) patients. From October 2000 to May 2002, 140 patients were enrolled. The treatment dose was P 66 mg m(-2) intravenous infusion (i.v.) on days 1, 8, and 15, and C 60 mg m(-2) i.v. on day 15, or V 23 mg m(-2) i.v. on days 1, 8, and 15, and C 60 mg m(-2) i.v. on day 15, every 4 weeks. In all, 281 cycles of PC and 307 cycles of VC were given to the patients in the PC and VC arms, respectively. There were 26 partial responses and one complete response (overall 38.6%) in the PC arm, and no complete responses, but 27 partial responses (overall 38.6%) in the VC arm. Myelosuppression was more common in the VC arm (P<0.001). Peripheral neuropathy and myalgia were significantly more common in the PC arm (P<0.001). The median time to disease progression was 6 months in the PC arm and 8.4 months in the VC arm (P=0.0344). The median survival time was 11.7 months in the PC arm and 15.4 months in the VC arm (P=0.297). We concluded that weekly PC is not suggested for NSCLC patients due to the relatively shorter progression-free survival and more common nonhaematological toxicities. British Journal of Cancer (2004) 90, 359-365. doi:10.1038/sj.bjc.6601526 www.bjcancer.com PMID- 14735178 TI - A double-blind, randomised, controlled Phase II trial of Heliox28 gas mixture in lung cancer patients with dyspnoea on exertion. AB - Helium has a low density and the potential of reducing the work of breathing and improving alveolar ventilation when replacing nitrogen in air. A Phase II, double blind, randomised, prospective, controlled trial was undertaken to assess whether Heliox28 (72% He/28% O(2)) compared with oxygen-enriched air (72% N(2)/28% O(2)) or medical air (78.9% N(2)/21.1% O(2)) could reduce dyspnoea and improve the exercise capability of patients with primary lung cancer and dyspnoea on exertion (Borg >3). A total of 12 patients (seven male, five female patients, age 53-78) breathed the test gases in randomised order via a facemask and inspiratory demand valve at rest and while performing 6-min walk tests. Pulse oximetry (SaO(2)) was recorded continuously. Respiratory rate and dyspnoea ratings (Borg and VAS) were taken before and immediately post-walk. Breathing Heliox28 at rest significantly increased SaO(2) compared to oxygen-enriched air (96+/-2 cf. 94+/-2, P<0.01). When compared to medical air, breathing Heliox28 but not oxygen-enriched air gave a significant improvement in the exercise capability (P<0.0001), SaO(2) (P<0.05) and dyspnoea scores (VAS, P<0.05) of lung cancer patients. PMID- 14735180 TI - Do CA125 response criteria overestimate tumour response in second-line treatment of epithelial ovarian carcinoma? AB - Recent studies indicate that cancer antigen 125 (CA125) response criteria tend to overestimate a tumour reduction measured by standard WHO response criteria in recurrent epithelial ovarian carcinoma. The aim of the study was to validate the recently introduced GCIG (The Gynaecological Cancer Intergroup) CA125 response criteria in predicting a tumour response measured by WHO (World Health Organization) criteria. Changes in CA125 levels (GCIG criteria) were retrospectively compared with alterations in the tumour load (WHO criteria) during second-line chemotherapy with topotecan or paclitaxel-platinum in 124 consecutive patients with recurrent or refractory disease. In patients assessable by both response criteria (n=72), the overall response rate using GCIG CA125 criteria was 57% (95% confidence interval (CI): 45-69%) and significantly higher than the response rate of 39% (95% CI: 28-51%) using WHO response criteria (P=0.045). The GCIG CA125 criteria had a sensitivity of 96% (95% CI: 82-100%), a specificity of 68% (95% CI: 52-81%) and an accuracy of 79% (95% CI: 68-88%) in predicting a response measured by WHO criteria. In conclusion, the GCIG CA125 response criteria seem to overestimate a tumour response by WHO criteria when monitoring the efficacy of second-line chemotherapy with topotecan or paclitaxel platinum in patients with epithelial ovarian carcinoma. PMID- 14735179 TI - Primary mediastinal large B-cell lymphoma (PMLBCL): long-term results from a retrospective multicentre Italian experience in 138 patients treated with CHOP or MACOP-B/VACOP-B. AB - The optimal treatment of primary mediastinal large B-cell lymphoma (PMLBCL) is still undefined. In the absence of randomised studies, we retrospectively analysed: (a) the effectiveness of two chemotherapy regimens (CHOP vs MACOP B/VACOP-B) in complete remission (CR) achievement and event-free survival (EFS) and (b) the role of mediastinal involved-field radiotherapy (IF-RT) as consolidation. From 1982 to 1999, 138 consecutive patients affected by PMLBCL were treated in 13 Italian institutions with CHOP (43) or MACOP-B/VACOP-B (95). The two groups of patients were similar as regard to age, gender, presence of bulky mediastinal mass, pleural effusion, stage and international prognostic indexes category of risk. Overall, 75.5% of patients in CR received IF-RT as consolidation. Complete remission was 51.1% in the CHOP group and 80% in MACOP B/VACOP-B (P<0.001). Relapse occurred in 22.7% of CHOP- and in 9.2% of MACOP B/VACOP-B-treated patients (n.s.). Event-free patients were 39.5% in CHOP and 75.7% in the MACOP-B/VACOP-B group (P<0.001). The addition of IF-RT as consolidation improved the outcome, irrespectively of the type of chemotherapy (P=0.04). At a multivariate analysis, achievement of CR (P<0.0001) and type of CT (MACOP-B/VACOP-B) retained the significance for OS (P=0.008) and EFS (P=0.03). In our experience, MACOP-B/VACOP-B appears to positively influence OS and EFS in patients affected by PMLBCL, as compared to CHOP. Consolidation IF-RT on mediastinum further improves the outcome of CR patients. PMID- 14735181 TI - Cost-effectiveness of stereotactic large-core needle biopsy for nonpalpable breast lesions compared to open-breast biopsy. AB - This paper demonstrates that the introduction of large-core needle biopsy (LCNB) replacing needle-localised breast biopsy (NLBB) for nonpalpable (screen-detected) breast lesions could result in substantial cost savings at the expense of a possible slight increase in breast cancer mortality. The cost-effectiveness of LCNB and NLBB was estimated using a microsimulation model. The sensitivity of LCNB (0.97) and resource use and costs of LCNB and NLBB were derived from a multicentre consecutive cohort study among 973 women who consented in getting LCNB and NLBB, if LCNB was negative. Sensitivity analyses were performed. Replacing NLBB with LCNB would result in approximately six more breast cancer deaths per year (in a target population of 2.1 million women), or in 1000 extra life-years lost from breast cancer (effect over 100 years). The total costs of management of breast cancer (3% discounted) are estimated at pound 4676 million with NLBB; introducing LCNB would save pound 13 million. The incremental cost effectiveness ratio of continued NLBB vs LCNB would be pound 12 482 per additional life-year gained (3% discounted); incremental costs range from pound 21 687 (low threshold for breast biopsy) to pound 74 378 (high sensitivity of LCNB). PMID- 14735182 TI - Breast density as a determinant of interval cancer at mammographic screening. AB - The association of breast density (% of breast volume involved by fibro-glandular densities) with the risk of interval cancer (IC) was investigated by reviewing a consecutive series of 346 cancers detected at screening (SDC) during 1996-1999 and of 90 ICs, reported as negative in the same period and diagnosed in the following 2 years, and comparing them to a random sample of 360 healthy controls. The probability of IC was significantly associated with breast density, whatever grouping (0/1-25/26-74/>74%; 0-25/26-60/61-74/>74%; 0-25/26-74/>74%) was considered (chi(2)=30.67-34.08, P<0.<0.01): 27.8% of all ICs were classified in the >74% density class, as compared to 7% of SDC and 5% of healthy controls. No significant association to IC was observed for Wolfe pattern (P2/Dy vs N1/P1: chi(2)=0.30, P=0.960), number of used mammographic views (single oblique vs oblique+craniocaudal: chi(2)=0.02, P=0.90) or screening round (first vs repeat: chi(2)=1.41, P=0.23). Multivariate analysis confirmed the independent association of breast density to IC, the highest risk being observed for >74% density class (OR vs 0% class=13.4, 95% CI 2.7-65.6, OR vs all other density classes=5.1, 95% CI 2.6-10.0). Age showed an independent association too, older women having a lower risk of IC (OR=0.52 95% CI 0.3-09). Breast density (>74%) resulted as being a major determinant of IC. Special screening protocols (shorter rescreening interval, routine use of ultrasonography) might be suggested for these subjects in order to improve screening sensitivity and efficacy. PMID- 14735183 TI - Economics of the clinical management of lung cancer in France: an analysis using a Markov model. AB - To evaluate, according to the histologic type and initial stage, the mean cost (MC) of managing patients with lung cancer and the costs of the different management phases. A Markov approach was used to model these costs, based on the management of a representative nation-wide sample of 428 patients with newly diagnosed lung cancer. The 18-month MC ranged from US$ 20 691 (95% CI: 5777-50 380 for diffuse non-small-cell lung cancer (NSCLC) to US$ 31 833 (95% CI: 15 866 64 455) for localised small-cell lung cancer (SCLC); first-line treatment costs ranged from 33.8% of MC for medically inoperable localised NSCLC to 74.6% for diffuse SCLC; second- or third-line treatment costs ranged from 7.8% of MC for surgically treated localised NSCLC to 32% for locally advanced NSCLC; and the cost of palliative care ranged from 9.1% of MC for locally advanced NSCLC to 39.9% for medically inoperable localised NSCLC. The cost of first-line chemotherapy and the percentage of actively treated patients impacted more on MC than did the cost of second- or third-line chemotherapy regimens or the cost of palliative care. In conclusion, this model provides a robust economic analysis of the cost of lung cancer management, and will be useful for assessing the economic consequences of future changes in patient management. PMID- 14735184 TI - Improved survival in patients with peritoneal metastases from colorectal cancer: a preliminary study. AB - Patients with peritoneal or local metastases from colorectal cancer have a poor prognosis. However, aggressive treatments by debulking surgery and infusional intraperitoneal (i.p.) chemotherapy have been tried and appear to benefit selected patients. We assayed the effects of debulking surgery and i.p. chemotherapy with respect to survival and compared the results with matched control patients treated by intravenous (i.v.) chemotherapy. In all, 18 patients with peritoneal and/or local metastases from colorectal adenocarcinoma underwent debulking surgery followed by 5-fluorouracil (5-FU) 550 mg m(-2) day(-1) i.p. and leucovorin (LV) 60 mg m(-2) day(-1) i.v. The chemotherapy was started the day after surgery and was given daily for 6 days and repeated monthly for totally eight courses. The control patients, matched for age, gender, performance status and metastatic site, were randomly selected from controlled clinical chemotherapy trials and treated with i.v. 5-FU+LV or i.v. methotrexate+5-FU+LV. There was no treatment-related mortality. The median survival among i.p. patients was 32 months compared to 14 months in the control group. In all, 11 patients who underwent macroscopically radical surgery had a longer survival than those who were not radically operated (P=0.02). These results indicate that patients with peritoneal metastases and/or locally advanced cancers but without distant metastases may benefit from cytoreductive surgery combined with i.p. chemotherapy. PMID- 14735185 TI - Potential health risks of complementary alternative medicines in cancer patients. AB - Many cancer patients use complementary alternative medicines (CAMs) but may not be aware of the potential risks. There are no studies quantifying such risks, but there is some evidence of patient risk from case reports in the literature. A cross-sectional survey of patients attending the outpatient department at a specialist cancer centre was carried out to establish a pattern of herbal remedy or supplement use and to identify potential adverse side effects or drug interactions with conventional medicines. If potential risks were identified, a health warning was issued by a pharmacist. A total of 318 patients participated in the study. Of these, 164 (51.6%) took CAMs, and 133 different combinations were recorded. Of these, 10.4% only took herbal remedies, 42.1% only supplements and 47.6% a combination of both. In all, 18 (11.0%) reported supplements in higher than recommended doses. Health warnings were issued to 20 (12.2%) patients. Most warnings concerned echinacea in patients with lymphoma. Further warnings were issued for cod liver/fish oil, evening primrose oil, gingko, garlic, ginseng, kava kava and beta-carotene. In conclusion, medical practitioners need to be able to identify the potential risks of CAMs. Equally, patients should be encouraged to disclose their use. Also, more research is needed to quantify the actual health risks. PMID- 14735186 TI - Polo-like kinase 1 overexpression is an early event in the progression of papillary carcinoma. AB - Polo-like kinase 1 (PLK1) is one of the serine threonine kinases that contributes to cell mitosis and is regarded as a marker of cellular proliferation. However, its protein expression in human carcinoma has not been studied in depth. We investigated PLK1 expression in various thyroid neoplasms in order to elucidate its physiological significance in thyroid carcinoma. Normal follicular cells only occasionally expressed PLK1. In follicular tumours and anaplastic carcinoma, PLK1 overexpression was not a common event and only 5.9% of follicular adenoma, 7.1% of follicular carcinoma, and 11.8% of anaplastic carcinoma overexpressed this protein. However, 43.7% of papillary carcinoma overexpressed PLK1. Polo-like kinase 1 overexpression was more frequently observed in smaller papillary carcinoma lesions, and 62.5% of microcarcinoma (ranging from 4 mm to 1.0 cm) and even 66.7% of incidental carcinoma (less than 4 mm) overexpressed it, whereas this phenomenon could only be seen in 20.0% of lesions larger than 4.0 cm. Furthermore, PLK1 overexpression was not related to cell-proliferating activity evaluated by Ki-67 labelling index, but it was inversely linked to UICC stage, extrathyroidal invasion, and the presence of poorly differentiated lesion as proposed by Sakamoto et al. These findings strongly suggest that, unlike other carcinomas previously studied, PLK1 does not act as a cell cycle regulator but plays a constitutive role in papillary carcinoma especially in the early phase, and may contribute to the malignant transformation of this carcinoma. PMID- 14735187 TI - The -174 G/C gene polymorphism in interleukin-6 is associated with an aggressive breast cancer phenotype. AB - Serum and tissue levels of interleukin-6 (IL-6) have been implicated in the biological phenotype of breast carcinoma. A common G/C polymorphism at position 174 of the IL-6 promoter can influence the expression level of this gene. We therefore investigated for associations between this polymorphism and various phenotypic features in a series of 256 breast cancers. Individuals who were homozygous for the C allele (n=55) were more likely to have higher-grade tumours (P=0.039) with ductal histology (P=0.030) compared to those harbouring at least one wild-type G allele (n=201). Homozygosity for the C allele was also associated with significantly worse overall survival (P=0.031). We conclude that the -174 C allele of IL-6 is associated with a more aggressive breast cancer phenotype. PMID- 14735188 TI - COX-2 expression is associated with an aggressive phenotype in ductal carcinoma in situ. AB - Cyclooxygenase type-2 (COX-2) is overexpressed in malignant tumours including breast cancers, though the mechanism of upregulation is unclear. This study aimed to determine COX-2 expression in ductal carcinoma in situ (DCIS) in comparison to invasive breast cancer (IBC) and normal breast, and also to investigate the relationship of COX-2 expression with HER-2 expression, oestrogen receptor (ER), tumour grade and cellular proliferation (Ki67) in DCIS. Cyclooxygenase type-2, HER-2, ER and Ki67 expression were determined by immunohistochemistry on paraffin tissue sections of DCIS (n=187), IBC (n=65) and normal breast reduction tissue (n=60). Cyclooxygenase type-2 expression in DCIS (67%, P<0.001) and IBC (63%, P<0.001) was significantly greater than in normal breast (23%). There was no difference in COX-2 expression level between DCIS and IBC (P=0.87) or between normal breast from reduction mammoplasty tissue and normal breast ducts around DCIS (22%, P=0.29). In DCIS, COX-2 expression was associated with higher cellular proliferation rates (P<0.0001), nuclear grade (P=0.003), with ER negativity (P=0.003) and with HER-2 positivity (P<0.0001). Cyclooxygenase type-2 expression is upregulated in in situ breast cancer and is associated with surrogate markers of an aggressive DCIS phenotype including nonoestrogen-regulated signalling pathways. Cyclooxygenase type-2 inhibition may potentially prevent the development of ER-positive and ER-negative breast cancers. PMID- 14735189 TI - Overexpression of VEGF121, but not VEGF165 or FGF-1, improves oxygenation in MCF 7 breast tumours. AB - Vascular endothelial growth factor (VEGF) is an intensively studied molecule that has significant potential, both in stimulating angiogenesis and as a target for antiangiogenic approaches. We utilised MCF-7 breast cancer cells transfected with either of two of the major VEGF isoforms, VEGF(121) or VEGF(165), or fibroblast growth factor-1 (FGF-1) to distinguish the effects of these factors on tumour growth, vascular function, and oxygen delivery. While each transfectant demonstrated substantially increased tumorigenicity and growth rate compared to vector controls, only VEGF(121) produced a combination of significantly reduced total and perfused vessel spacing, as well as a corresponding reduction in overall tumour hypoxia. Such pathophysiological effects are of potential importance, since antiangiogenic agents designed to block VEGF isoforms could in turn result in the development of therapeutically unfavourable environments. If antiangiogenic agents are also combined with conventional therapies such as irradiation or chemotherapy, microregional deficiencies in oxygenation could play a key role in ultimate therapeutic success. PMID- 14735190 TI - Low expression of polypeptide GalNAc N-acetylgalactosaminyl transferase-3 in lung adenocarcinoma: impact on poor prognosis and early recurrence. AB - Initial glycosylation of mucin-type O-linked protein is catalysed by one of the UDP-GalNAc: polypeptide N-acetyl-galactosaminyl transferase-3 (GalNAc-T3). O glycosylation is important in the binding of cell adhesion molecules, cell differentiation, invasion, and metastasis in tumours. This study was designed to detect GalNAc-T3 expression in lung adenocarcinoma by using immunohistochemical staining, and to evaluate the relationship between the GalNAc-T3 expression level and prognosis and recurrence in completely resected lung adenocarcinoma patients. A low expression of GalNAc-T3 was detected in the cytoplasm of tumour cells in 79 of 148 patients (53.4%) with lung adenocarcinoma. The low expression of GalNAc-T3 was associated with poorly differentiated tumour (P<0.0001), poor pathologic stage (P<0.0001), lymph node metastasis (P<0.0001), and tumour recurrence (P=0.016). The lung carcinoma patients with low GalNAc-T3 expression had a poorer prognosis than those with high GalNAc-T3 expression, using both univariate and multivariate analyses (overall survival: P<0.0001 and P=0.011, respectively). In addition, multivariate analysis of the clinicopathological characteristics of stage I lung adenocarcinoma indicated that the low expression of GalNAc-T3 was a significant independent factor for predicting poor prognosis and early recurrence (P=0.006, rr=2.87 and P=0.019, rr=3.05, respectively). The low expression of GalNAc-T3 may be a useful marker for predicting poor prognosis and early recurrence in completely resected lung carcinoma patients, particularly patients with stage I diseases. PMID- 14735191 TI - Detection of HER-2/neu-positive circulating epithelial cells in prostate cancer patients. AB - HER-2/neu may play a role in prostate carcinogenesis. The aim of this study was to use the expression of HER-2/neu as a molecular marker for the detection of circulating tumour cells (CTCs) in the blood of patients with prostate cancer (PC). Blood samples were collected from 42 patients with PC and nine healthy volunteers. Immunomagnetic beads were used to harvest epithelial cells from peripheral blood mononuclear cells. Total RNA was extracted and reverse transcribed before analysis by real-time PCR with HER-2/neu-specific primers. CTCs were HER-2/neu positive in six out of 11 (54%) patients with metastatic disease and in three out of 31 (9.6%) patients with localised PC (P=0.004). In blood samples from nine healthy volunteers, we detected no expression of HER 2/neu. The present method appears to be minimally invasive, highly sensitive and a specific approach for detecting CTCs in PC. Furthermore, it may help better target HER-2/neu in advanced PC. PMID- 14735192 TI - Expression of the epidermal growth factor receptor family in prostate carcinoma before and during androgen-independence. AB - Novel palliative strategies for patients with androgen-independent prostate cancer (AIPC) include targeting the epidermal growth factor receptor (EGFR) family. The aim of the present study was to investigate intrapatient changes of EGFRs during the development of AIPC. In total, 106 symptomatic AIPC patients were identified in whom prostatic biopsies (adenocarcinoma) were available both before the start of androgen deprivation (PRTR biopsy) and after the development of AIPC (AIPC biopsy). All four known subgroups of the EGFR family were determined by immunohistochemistry (IHC): c-erbB-1 (EGFR), c-erbB-2 (HER2/neu), c erbB-3 (HER3) and c-erbB-4 (HER4). Moderate to strong membrane-specific staining was recorded semiquantitatively (<10% vs >/=10%=IHC stained tumour cells: 'negative' vs 'positive' staining). The medical records were reviewed for clinical variables. During the development of AIPC, intrapatient changes occurred in two opposite directions for each of the four EGFRs: negativity changed to positivity, and vice versa, statistically significant only for the increase of c erbB-1 expression (P=0.001). The c-erbB-2 expression in the AIPC biopsy was associated with a significantly shorter survival from the time of the AIPC biopsy (P=0.029). Our results support ongoing therapeutic attempts of EGFR inhibition in subgroups of patients with prostate cancer. Further research is needed to understand the function of EGFRs in this malignancy. PMID- 14735193 TI - Expression of MTA1 promotes motility and invasiveness of PANC-1 pancreatic carcinoma cells. AB - The human metastasis-associated protein 1 (MTA1) is a constituent of the nucleosome-remodelling and -deacetylation complex. Its expression has been correlated with the invasion and metastasis of epithelial neoplasms. To address the functional consequences of MTA1 expression in pancreatic carcinoma cells, we have established PANC-1 pancreatic carcinoma cells that stably express MTA1 as an enhanced green fluorescent fusion protein (EGFP-MTA1). Here, we demonstrate that heterologous expression of EGFP-MTA1 markedly enhanced the cellular motility and the invasive penetration of epithelial barriers by the cells. Expression of EGFP MTA1 had no effect on substrate-independent growth, but reduced substrate dependent cell proliferation. In addition, the organisation of the cytokeratin filament system and the localisation of the actin cytoskeleton-associated protein IQGAP1 were distinctly altered in EGFP-MTA1-expressing cells. These results indicate that enhanced expression of MTA1 promotes the acquisition of an invasive, metastatic phenotype, and thus enhances the malignancy of pancreatic adenocarcinoma cells by modulation of the cytoskeleton. PMID- 14735194 TI - Tissue inhibitor of metalloproteinase-1 stimulates proliferation of human cancer cells by inhibiting a metalloproteinase. AB - TIMP-1, an approximately 30 kDa glycosylated protein found predominantly in extracellular compartments, is involved in the regulation of a variety of developmental, remodelling, and pathological processes. One function of TIMP-1 is to inhibit certain members of a group of extracellular and cell surface enzymes known collectively as metalloproteinases (MP). These include the matrix metalloproteinases and the adamalysin-like disintegrin and metalloproteinases (ADAMs). Additional activities of TIMP-1 include potentiating the activity of erythroid precursors and stimulating proliferation of certain cancer cell lines. Published evidence suggests that the apparent proliferative action of TIMP-1 is independent of its MP-inhibitory activity; however, reports of a cell surface receptor for TIMP-1 have not been confirmed. We have utilised a baculovirus-based system to produce TIMP-1. Data presented here show that TIMP-1 and synthetic hydroxamate (GM6001) MP inhibitors stimulate proliferation and metabolic activity of MDA-MB-435 cancer cells with similar kinetics. An inactive hydroxamate derivative was ineffective. The TIMP-1-induced increase in proliferation and metabolic activity was not the consequence of the inhibition of apoptosis by TIMP 1 in the serum-free medium. These data taken together imply that the mechanism by which TIMP-1 enhances cell growth depends on its ability to inhibit a metalloproteinase, rather than to stimulate a cell surface receptor by a process independent of its MP-inhibitory activity. Inhibitors of extracellular regulated kinase (U0126) and p38 (SB203580), and to a lesser extent the phosphatidylinositol-3-kinase inhibitor LY294002, suppressed the action of TIMP 1. Assays for ERK1/2 and p38 showed that both were activated by TIMP-1 and GM6001. Mechanisms by which TIMP-1 might act to stimulate cell proliferation are described. PMID- 14735195 TI - Motility-related protein-1 (MRP-1/CD9) expression can predict disease-free survival in patients with squamous cell carcinoma of the head and neck. AB - CD9 is a transmembrane protein that has been implicated in cell adhesion, motility and proliferation, and numerous studies have demonstrated the prognostic value of its expression in different solid tumours. The purpose of this study is to determine the predictive value of CD9 in squamous cell carcinoma (SCC) of the head and neck. A total of 153 cases were examined for CD9 expression using immunohistochemistry applied on formalin-fixed, paraffin-embedded tissue. Cases were stratified in two categories depending on CD9 expression, as positive (>/=50% positive cells) or reduced (<50%). In all, 108 cases were positive for CD9 (85 cases with membranous, and 23 with both membranous and cytoplasmic staining) and 45 reduced expression. Reduced CD9 expression was significantly associated with high grade (P=0.0007) and lower disease-free survival (DFS) (P=0.017). The latter retained its significance in the multivariate analysis. When the 23 cases with both membranous and cytoplasmic patterns were studied as a separate subgroup, there were significant associations between CD9 expression and tumour grade (P=0.025) (95% CI 11-68), tumour stage (P=0.08) (95% CI 3.5-86) and the occurrence of any failure (P=0.083) (95% CI -1.7-57). Immunohistochemical CD9 expression proved to be an independent prognostic factor in SCC of the head and neck, and it may detect patients at a high risk of recurrence. In addition, the cytoplasmic pattern seems to have an even more significant value. However, this finding is limited to the small number of cases with this pattern. PMID- 14735196 TI - Variation in RNA expression and genomic DNA content acquired during cell culture. AB - Specific chromosomal abnormalities are increasingly recognised to be associated with particular tumour subtypes. These cytogenetic abnormalities define the sites of specific genes, the alteration of which is implicated in the neoplastic process. We used comparative genomic hybridisation (CGH) to examine DNA from different breast and ovarian cancer cell lines for variations in DNA sequence copy number compared with the same normal control. We also compared different sources of the MCF7 breast line by both CGH and cDNA expression arrays. Some of the differences between the subcultures were extensive and involved large regions of the chromosome. Differences between the four subcultures were observed for gains of 2q, 5p, 5q, 6q, 7p, 7q, 9q, 10p, 11q, 13q, 14q, 16q, 18p and 20p, and losses of 4q, 5p, 5q, 6q, 7q, 8p, 11p, 11q, 12q, 13q, 15q, 19p, 19q, 20p, 21q, 22q and Xp. However, few variations were found between two subcultures examined, 5 months apart, from the same initial source. The RNA arrays also demonstrated considerable variation between the three different subcultures, with only 43% of genes expressed at the same levels in all three. Moreover, the patterns of the expressed genes did not always reflect our observed CGH aberrations. These results demonstrate extensive genomic instability and variation in RNA expression during subculture and provide supportive data for evidence that cell lines do evolve in culture, thereby weakening the direct relevance of such cultures as models of human cancer. This work also reinforces the concern that comparisons of published analyses of cultures of the same name may be dangerous. PMID- 14735198 TI - Chromosomal imbalances associated with anaplastic transformation of follicular thyroid carcinomas. AB - The genetic alterations that underlie the progression of follicular thyroid carcinoma towards anaplasia are still largely uncharacterised. We compared the Comparative Genomic Hybridization (CGH) profiles of 20 follicular (FTCs), 12 poorly differentiated (PDTCs) and seven anaplastic thyroid carcinomas (ATCs), in order to identify the chromosomal imbalances potentially associated with cancer progression. We found: (i) when considering that a 'direct' transformation of FTC towards anaplasia occurs, the defined significantly important alterations were the increase of gains at 3q (P<0.05) and 20q (P<0.01), and the increase of losses at 7q (P<0.05) and Xp (P<0.01); (ii) regarding poorly differentiated carcinomas as an intermediate independent entity in the anaplastic transformation of follicular cancers, evidenced as important alterations towards anaplasia, were the proportional decrease in copy sequences at 7p, 7q, 12q and 13q resulting from the significant decrease of DNA gains at 7p and 12q (P<0.05), and the significant increase of losses at 7q and 13q (P<0.05). These results unveil the chromosomal regions where genes of interest in thyroid anaplastic transformation are to be located, and demonstrate that different gene dosage copy sequence imbalances are associated to the 'direct' pathway of transformation of follicular into anaplastic cancers and to the progressive FTC --> PDTC --> ATC pathway. PMID- 14735197 TI - Germline truncating mutations in both MSH2 and BRCA2 in a single kindred. AB - There has been interest in the literature in the possible existence of a gene that predisposes to both breast cancer (BC) and colorectal cancer (CRC). We describe the detailed characterisation of one kindred, MON1080, with 10 cases of BC or CRC invasive cancer among 26 first-, second- or third-degree relatives. Linkage analysis suggested that a mutation was present in BRCA2. DNA sequencing from III: 22 (diagnosed with lobular BC) identified a BRCA2 exon 3 542G>T (L105X) mutation. Her sister (III: 25) had BC and endometrial cancer and carries the same mutation. Following immunohistochemical and microsatellite instability studies, mutation analysis by protein truncation test, cDNA sequencing and quantitative real-time PCR revealed a deletion of MSH2 exon 8 in III: 25, confirming her as a double heterozygote for truncating mutations in both BRCA2 and MSH2. The exon 8 deletion was identified as a 14.9 kb deletion occurring between two Alu sequences. The breakpoint lies within a sequence of 45 bp that is identical in both Alu sequences. In this large BC/CRC kindred, MON1080, disease-causing truncating mutations are present in both MSH2 and BRCA2. There appeared to be no increased susceptibility to the development of colorectal tumours in BRCA2 mutation carriers or to the development of breast tumours in MSH2 mutation carriers. Additionally, two double heterozygotes did not appear to have a different phenotype than would be expected from the presence of a mutation in each gene alone. PMID- 14735199 TI - XPD gene polymorphism and host characteristics in the association with cutaneous malignant melanoma risk. AB - We recently reported an association between low DNA repair capacity, measured through the host-cell reactivation assay, and melanoma risk in subjects with dysplastic naevi or low tanning ability. We investigated the genetic basis for these findings by analysing the Asp312Asn and Lys751Gln polymorphisms of the XPD (ERCC2) DNA repair gene in the same subjects. Similar to our previous report, no significant association between XPD polymorphisms and melanoma risk was found in 176 melanoma cases and 177 controls (odds ratio (OR)=1.5, 95% confidence interval (CI)=0.9-2.5 for 312Asn; OR=1.3, 95% CI=0.8-2.1 for 751Gln, adjusted for age, gender, dysplastic naevi and pigmentation characteristics). However, XPD variants were associated with increased risk in older (>50 years) subjects (OR=3.4, 95% CI=1.6-7.3 for 312Asn; OR=2.3, 95% CI=1.1-4.9 for 751Gln). The 751Gln allele was associated with elevated melanoma risk among subjects without dysplastic naevi (OR=2.6, 95% CI=1.1-6.4). Subjects with low tanning ability and XPD variants exhibited a nonsignificant increase of melanoma risk (OR=2.3, 95% CI=0.7-7.0 for 312Asn; OR=3.0, 95% CI=1.0-8.8 for 751Gln). DNA repair capacity was slightly decreased in subjects carrying 751Gln alleles. XPD variants may modify melanoma risk in subjects with specific host characteristics, such as older age, lack of dysplastic naevi or low tanning ability. PMID- 14735200 TI - Germline mutations of the INK4a-ARF gene in patients with suspected genetic predisposition to melanoma. AB - Germline anomalies of the INK4a-ARF and Cdk4 genes were sought in a series of 89 patients suspected of having a genetic predisposition to melanoma. Patients were selected based on the following criteria: (a) familial melanoma (23 cases), (b) multiple primary melanoma (MPM; 18 cases), (c) melanoma and additional unrelated cancers (13 cases), (d) age at diagnosis less than 25 years (21 cases), and (e) nonphoto-induced melanoma (NPIM; 14 cases). Mutations of INK4a-ARF and Cdk4 were characterised by automated sequencing, and germline deletions of INK4a-ARF were also examined by real-time quantitative PCR. Seven germline changes of INK4a-ARF, five of which were novel, were found in seven patients (8%). Four were very likely to be pathogenic mutations and were found in three high-risk melanoma families and in a patient who had a pancreatic carcinoma in addition to melanoma. Three variants of uncertain significance were detected in one MPM patient, one patient <25 years, and one NPIM patient. No germline deletion of INK4a-ARF was found in 71 patients, and no Cdk4 mutation was observed in the 89 patients. This study confirms that INK4a-ARF mutations are infrequent outside stringent familial criteria, and that germline INK4a-ARF deletions are rarely involved in genetic predisposition to melanoma. PMID- 14735201 TI - Hereditary prostate cancer in African American families: linkage analysis using markers that map to five candidate susceptibility loci. AB - African American men have the highest incidence of prostate cancer in the world. Despite this statistic, linkage studies designed to localise prostate cancer susceptibility alleles have included primarily men of Caucasian descent. In this report, we performed a linkage analysis using 33 African American prostate cancer families from two independent research groups. In total, 126 individuals (including 89 men with prostate cancer) were genotyped using markers that map to five prostate cancer susceptibility loci, namely HPC1 at 1q24-25, PCAP at 1q42.2 43, CAPB at 1p36, HPC20 on chromosome 20, and HPCX at Xq27-28. Multipoint mode-of inheritance-free linkage analyses were performed using the GENEHUNTER software. Some evidence of prostate cancer was detected to HPC1 using all families with a maximum NPL Z score of 1.12 near marker D1S413 (P=0.13). Increased evidence of linkage was observed in the 24 families with prostate cancer diagnosis prior to age 65 years and in the 20 families with male-to-male transmission. Some evidence of prostate cancer linkage was also detected at markers mapping to PCAP, HPC20, and HPCX. Continued collection and analysis of African American prostate cancer families will lead to an improved understanding of inherited susceptibility in this high-risk group. PMID- 14735202 TI - SLIT2 promoter methylation analysis in neuroblastoma, Wilms' tumour and renal cell carcinoma. AB - The 3p21.3 RASSF1A tumour suppressor gene (TSG) provides a paradigm for TSGs inactivated by promoter methylation rather than somatic mutations. Recently, we identified frequent promoter methylation without somatic mutations of SLIT2 in lung and breast cancers, suggesting similarities between SLIT2 and RASSF1A TSGs. Epigenetic inactivation of RASSF1A was first described in lung and breast cancers and subsequently in a wide range of human cancers including neuroblastoma, Wilms' tumour and renal cell carcinoma (RCC). These findings prompted us to investigate SLIT2 methylation in these three human cancers. We analysed 49 neuroblastomas (NBs), 37 Wilms' tumours and 48 RCC, and detected SLIT2 promoter methylation in 29% of NB, 38% of Wilms' tumours and 25% of RCC. Previously, we had demonstrated frequent RASSF1A methylation in the same tumour series and frequent CASP8 methylation in the NB and Wilms' tumour samples. However, there was no significant association between SLIT2 promoter methylation and RASSF1A or CASP8 methylation in NB and RCC. In Wilms' tumour, there was a trend for a negative association between RASSF1A and SLIT2 methylation, although this did not reach statistical significance. No associations were detected between SLIT2 promoter methylation and specific clinicopathological features in the tumours analysed. These findings implicate SLIT2 promoter methylation in the pathogenesis of both paediatric and adult cancers and suggest that further investigations of SLIT2 in other tumour types should be pursued. However, epigenetic inactivation of SLIT2 is less frequent than RASSF1A in the tumour types analysed. PMID- 14735203 TI - Molecular variants of the ATM gene in Hodgkin's disease in children. AB - Ataxia telangiectasia is an autosomal recessive disease with a striking predisposition of lymphoid malignancies. ATM mutations have been reported in adult sporadic lymphoma and leukaemia. The aim of this study was to investigate the possible involvement of the ATM gene in the carcinogenesis of Hodgkin disease in children. Tumours were obtained from 23 patients and were subjected to mutation screening and loss of heterozygosity analysis. Eight base substitutions were identified in seven patients. Of them, Y54Y, a silent change, was observed in two patients and a known polymorphism, D1853N, in three patients. Of the other two patients, one harboured a combined genotype P604S/F1463C, identified previously in two patients with Hodgkin lymphoma, and the other a novel missense mutation, V595A. The alterations were present in the germ line, and both had a more aggressive disease. In all, 100 matched normal ethnic controls were screened for these mutations and P604S/F1463C was identified in one healthy control. Loss of heterozygosity was identified in four patients and in three of them it was located centromeric to the ATM gene, and, in one, it spanned a large region, indicating the involvement of other tumour-suppressor genes in this disease. Missense variants of the ATM gene are a rare event in childhood Hodgkin disease. PMID- 14735204 TI - Thymidylate synthase and methylenetetrahydrofolate reductase gene polymorphisms: relationships with 5-fluorouracil sensitivity. AB - The relationship of thymidylate synthase (TS) and methylenetetrahydrofolate reductase (MTHFR) gene polymorphisms on 5-fluorouracil (FU) sensitivity was tested on 19 human cancer cell lines (head and neck, breast, digestive tract) in the absence and presence of folinic acid (FA) supplementation. Thymidylate synthase polymorphisms in the 5' promoter region (double or triple tandem repeats) and 3' untranslated region (6-bp deletion) were analysed by PCR. The C677T and A1298C MTHFR polymorphisms were determined by melting curve analyses (LightCycler). Thymidylate synthase activity and intracellular concentration of the reduced folate 5-10 methylenetetrahydrofolate (CH(2)FH(4)) were measured (biochemical assays). Thymidylate synthase activity was significantly different according to 5' TS genotype, heterozygous cell lines (2R/3R) exhibiting higher TS activities than homozygous ones (P=0.05). However, whether in the absence or presence of FA, FU sensitivity was not statistically associated with either 5' or 3' TS polymorphism. Basal CH(2)FH(4) cellular concentrations were lowest in C677T homozygous wild-type (wt) (C/C) cell lines. FU sensitivity was not linked to C677T polymorphism. In contrast, there was a marked trend for a greater FU efficacy in mutated A1298C variants (C/C+A/C) as compared to wt homozygous cell lines (A/A) (P=0.055 and 0.085 without and with FA supplementation, respectively). These results suggest for the first time a potential role of A1298C MTHFR polymorphism on fluoropyrimidine sensitivity. PMID- 14735205 TI - Sodium butyrate and tributyrin induce in vivo growth inhibition and apoptosis in human prostate cancer. AB - Histone deacetylase inhibitors (HDACs) are known to exhibit antiproliferative effects on various carcinoma cells. In this study, the in vivo efficiency of two HDACs, sodium butyrate and tributyrin, on prostate cancer growth inhibition were investigated. To gain an insight into the possible underlying pathways, cell culture experiments were performed focusing on the expression of p21, Rb and c myc. For in vivo testing, prostate cancer cell lines (PC3 and TSU-Pr1) were seeded on the chorioallantois membrane (CAM) and implanted in a xenograft model using nude mice. Standard Western blot analysis was performed for protein expression of p21, Rb and c-myc in HDAC-treated vs untreated prostate cancer cells. Both sodium butyrate and tributyrin had a considerable treatment effect on microtumours on the chicken egg at already very low concentrations of 0.1 mM. Tributyrin-treated tumours showed the strongest effect with 38% apoptotic nuclei in the prostate cancer cell line PC3. In the mouse model, there was almost no difference between sodium butyrate and tributyrin. In untreated animals the tumours were almost double the size 4 weeks after implantation. Tumours of the treatment groups had a significantly lower percentage of Ki-67-positive-stained nuclei. As demonstrated by Western blot analysis, these effects seem to be independent of p53 status and a pathway via p21-Rb-c-myc is possibly involved. In this study we have demonstrated a substantial in vivo treatment effect, which can be induced by the application of sodium butyrate or the orally applicable tributyrin in human prostate cancer. The given results may provide the rationale to apply these drugs in well-controlled clinical trials in patients being at high risk of recurrence after specific therapy or in patients with locally or distant advanced prostate cancer. PMID- 14735206 TI - An evaluation of gemcitabines differential radiosensitising effect in related bladder cancer cell lines. AB - The aim of this study was to establish the radiosensitising properties of gemcitabine in a pair of related bladder tumour cell lines with differential radiosensitivity. The radioresistant bladder tumour cell line MGH-U1 and its radiosensitive mutant clone, S40b (both p53 mutant), had SF(2) values (surviving fraction at 2 Gy) of 0.98 and 0.64, respectively (P<0.001). Colony-forming assays showed that at 0.01 microM gemcitabine radiosensitisation occurred only in the S40b cell line (dose-modifying factor (DMF)=1.4). At 0.3 microM (killing 50% of cells), both cell lines were radiosensitised; DMF=2.25 and 1.2 for MGH-U1 and S40b, respectively. These data suggest that gemcitabine is an effective radiosensitiser in bladder cancer cell lines, with greater sensitisation in the radioresistant parental line-a feature that should be useful in a clinical setting. PMID- 14735207 TI - Induction of tumour blood flow stasis and necrosis: a new function for epinephrine similar to that of combretastatin A-4 derivative AVE8062 (AC7700). AB - AVE8062, a derivative of combretastatin A-4, has a strong stanching effect on tumour blood flow (TBF), which leads to complete blockage of nutrient supply to solid tumours and their necrosis. Previously, we reported that TBF stasis is due to increased arteriolar resistance caused by AVE8062 and a lasting decrease in perfusion pressure in tumour-feeding vessels. Here, we measured changes in TBF in rat solid tumour LY80 during continuous administration of AVE8062-like epinephrine or four catecholamines that are unlike AVE8062 (norepinephrine, dopamine, methoxamine, and metaraminol) to the region of increased vascular resistance. Venous administration of 0.3 mg ml(-1) epinephrine caused TBF to fall immediately to near zero, where it remained throughout the administration period. With a 30-min drug administration, TBF began to recover immediately when drug administration halted. With a 60-min epinephrine administration, TBF recovered somewhat, but not to the previous level. With drug administration of 120 min, TBF did not recover during the subsequent 8 h. Likewise, 0.1 mg ml(-1) epinephrine produced irreversible occlusion after 120 min of administration. In contrast, 120 min of administration of the four other catecholamines resulted in no occlusion. Only the group given 0.3 mg ml(-1) epinephrine (not that given methoxamine) showed significantly greater necrosis than the control. We conclude that, for epinephrine to cause irreversible occlusion of these vessels, a marked decrease in perfusion pressure in tumour-feeding blood vessels is necessary and should be maintained for 2 h. This conclusion is consistent with the previously demonstrated mechanism of irreversible arteriole occlusion caused by AVE8062. AVE8062 and epinephrine appear to have the same mechanism of action regarding induction of tumour blood flow stasis. PMID- 14735208 TI - Cellular radiosensitivity: do separate predictive parameters apply for fibroblasts and for human tumour cells? PMID- 14735214 TI - A survey of Campylobacter species shed in faeces of beef cattle using polymerase chain reaction. AB - A polymerase chain reaction (PCR)-based survey of campylobacters associated with faeces collected from 382 beef cattle was undertaken. To ensure the removal of PCR inhibitors present in faeces and determine if adequate extraction was achieved, faeces were seeded with internal control DNA (i.e., DNA designed to amplify with the Campylobacter genus primer set, but provide a smaller amplicon) before the extraction procedure. In only two samples (0.5%) were the internal control or Campylobacter genus amplicons not detected. In the remaining 380 faecal samples, Campylobacter DNA was detected in 83% of the faecal samples (80% of the faecal samples were positive for Campylobacter genus DNA, and 3% of the samples were negative for Campylobacter genus DNA but positive for DNA of individual species). The most frequently detected species was Campylobacter lanienae (49%), a species only recently connected to livestock hosts. Campylobacter jejuni DNA was detected in 38% of the faecal samples, and Campylobacter hyointestinalis and Campylobacter coli DNA were detected in 8% and 0.5% of the samples, respectively. Campylobacter fetus DNA was not detected. Twenty-four percent of the faecal samples contained DNA of at least two species of Campylobacter. Of these samples, the majority (81%) contained DNA of C. jejuni and C. lanienae. The results of this study indicate that beef cattle commonly release a variety of Campylobacter species into the environment and may contribute to the high prevalence of campylobacteriosis in humans inhabiting areas of intensive cattle production, such as southern Alberta. Furthermore, this study demonstrates the utility of using PCR as a rapid and accurate method for simultaneously detecting the DNA of a diverse number of Campylobacter species associated with bovine faeces. PMID- 14735215 TI - Characteristics of the squalene synthase inhibitors produced by a Streptomyces species isolated from soils. AB - Microorganisms producing squalene synthase inhibitors were screened from soils. A high producer was selected and identified as a Streptomyces species. Two active inhibitors were obtained from culture broths via a series of purification processes involving solvent extraction, WK-10 cation-exchange column chromatography, HP-20 adsorption column chromatography, silica-gel column chromatography, preparative HPLC, and crystallization. The inhibitors were confirmed as macrolactins A and F with molecular weights of 402 by UV-absorption spectrometry, fast atom bombardment mass spectometry, and 13C- and 1H-NMR analyses. Kinetic results for macrolactins A and F showed that they appear to be noncompetitive inhibitors of rat liver squalene synthase with IC50 values of 1.66 and 1.53 micromol/L, respectively. Since mammalian squalene synthase was used, these inhibitors have significant potential as therapeutic agents for hyperlipemia and suppression of cholesterol biosynthesis. PMID- 14735216 TI - Screening of nonpolyenic antifungal metabolites produced by clinical isolates of actinomycetes. AB - The purpose of this work was to screen clinical isolates of actinomycetes producing nonpolyenic antifungals. This choice was made to limit the problem of rediscovery of well-known antifungal families, especially polyenic antifungals. One hundred and ten strains were tested, using two diffusion methods and two test media, against three yeast species and three filamentous fungi. Among 54 strains (49%) showing antifungal activity, five strains belonging to the genus Streptomyces were active against all test organisms and appeared promising. These results indicate that clinical and environmental isolates of actinomycetes could be an interesting source of antifungal bioactive substances. The production of nonpolyenic antifungal substances by these five active isolates was investigated using several criteria: antibacterial activity, ergosterol inhibition, and UV visible spectra of active extracts. One active strain responded to all three selection criteria and produced potentially nonpolyenic antifungal metabolites. This strain was retained for further investigation, in particular, purification, structure elucidation, and mechanism of action of the active product. PMID- 14735217 TI - Manganese-lignin peroxidase hybrid from Bjerkandera adusta oxidizes polycyclic aromatic hydrocarbons more actively in the absence of manganese. AB - We studied polycyclic aromatic hydrocarbon (PAH) oxidation using whole cells and purified manganese-lignin peroxidase (MnLiP) from Bjerkandera adusta UAMH 8258. Although the metabolism of PAHs by B. adusta has been previously demonstrated, less than 5% mineralization of 14C-labelled PAHs occurred in this study over a 40 day period. Oxidation of PAHs was examined by a purified MnLiP hybrid isoenzyme in the presence and absence of manganous ions. The rate of PAH oxidation was decreased by the presence of Mn. The substrates were anthracene and its methyl derivatives, pyrene and benzo[a]pyrene, PAHs with ionization potentials of 7.43 eV or lower. The PAH metabolites of the Mn-independent reaction were identified as the corresponding quinones. The pH optimum of the Mn-independent oxidation was generally about 4, while for the Mn-dependent reaction it was 3. The kinetic constants for the Mn-independent oxidation of 2-methylanthracene at pH 4 were determined, and the values we obtained were a kcat of 145/min, KM,app of 23.8 mmol/L for the aromatic substrate, and KM,app of 0.2 mmol/L for hydrogen peroxide. This is the first report of PAH oxidation by a MnLiP hybrid isoenzyme from white rot fungi. PMID- 14735218 TI - Actinobacterial chitinase-like enzymes: profiles of rhizosphere versus non rhizosphere isolates. AB - The objective of this study was to determine if antifungal actinomycetes isolated from rhizosphere and non-rhizosphere soils exhibit different chitinase-like production and (or) induction patterns. Selected isolates from both habitats were compared. Chitinase-like levels and isoform characteristic patterns were evaluated over time in culture fluids of isolates grown on media containing different combinations of colloidal chitin and fungal cell wall (FCW) preparation. Supernatants were also subjected to native and non-native polyacrylamide gel electrophoresis (PAGE), using glycol chitin amended gels. For non-native PAGE, protein samples were denatured by two different approaches. Multiple active bands, ranging from 20 to 53 kDa and present in varying amounts, were detected in gels for most strains. Different substrate preferences were observed among strains, and different chitinase-like enzymes were produced, depending upon the substrate combinations used. The presence of FCW in the medium induced specific chitinase-like enzymes not observed otherwise. Enzymatic activities and profiles of the isolates, however, were strain and substrate specific rather than habitat specific. However, a sagebrush rhizosphere soil had a larger actinomycete community with higher chitinolytic activities than the nearby bulk soil. The use of PAGE to compare chitinase-like proteins induced in media with and without FCW was useful for identifying chitinase-like enzymes potentially involved in antifungal activity. PMID- 14735219 TI - Microbial production of isoquinoline from indene. AB - A purified microbial isolate, identified as a strain of Rhodococcus sp., metabolized indene primarily to iso quinoline and lesser amounts of indandiol and indanone. Isoquinoline production was dependent on the presence of microbial culture, indene, and ammonium ions as the source of nitrogen in the molecule. The ability to produce isoquinoline was induced by growth on benzene or naphthalene and by the presence of indene itself. The culture produced compounds tentatively identified as 3-methylisoquinoline and 3-ethylisoquinoline from 2-methylindene and from 2-ethylindene, respectively. Deuterated indene was converted to deuterated isoquinoline, deuterated indanone, and deuterated indandiol. Experiments with [15N]ammonium nitrate and ammonium [15N]nitrate confirmed ammonium as the source of nitrogen in the isoquinoline products. PMID- 14735220 TI - Survival of lactococci during passage through mouse digestive tract. AB - One of the important properties of probiotics is the ability to survive in the intestine. There have been few studies on the probiotic property of lactococci, since they are formally not considered to be natural inhabitants of the intestine. To evaluate lactococci as probiotic bacteria, we investigated their ability to survive during gastric transit by in vitro and in vivo tests. When exposed to an in vitro simulated gastrointestinal environment, such as low pH and bile, only Lactococcus lactis subsp. lactis bv. diacetylactis N7 showed a moderate survival rate among the four strains tested. The tested strains were orally administered to mice, and intestinal passage of the ingested strains was monitored by two methods: antibiotics and PCR. Viable cells of strain N7 were recovered from feces within 24-48 h after administration but not at 72 h. Lactococcus lactis subsp. cremoris ATCC 19257, which had a poor survival rate in vitro test, was also detected at 12 h but not at 24 h. These results indicate that lactococci can reach the mouse intestine alive, but not colonize it. If administered daily, viable strain N7 may exist continuously in the intestine. The effect of strain N7 on intestinal microbial balance and on animal health will be the subject of a further study. PMID- 14735221 TI - Influence of high salinities on the degradation of diesel fuel by bacterial consortia. AB - Microbial communities from three Argentinean saline soils were extracted and tested for their ability to degrade diesel fuel in liquid culture at salinities between 0% and 25%. In each case, the degradation process was continuously monitored by measuring oxygen consumption. Two communities (CR1 and CR2) showed nearly equal degrees of degradation across a salinity range of 0%-10% (the former degrading about 63% of the diesel fuel and the latter about 70% after 53 and 80 d, respectively). Furthermore, the degree of degradation was not significantly lower in the presence of 17.5% salt (58% and 65% degraded, respectively). A third community (El Zorro) showed a maximum turnover at 5% salt (79% diesel fuel degraded) and significant degradation (66%) at a salinity of 10%. However, the degree of degradation by this community clearly dropped at 0% and 15% salt. None of the communities were able to degrade diesel fuel in the presence of 25% salt, but the living cell counts showed that components of the microbial population survived the long-term exposure. The surviving portion is obviously sufficient to allow substantial restoration of the original community, as verified by the BIOLOG method. Isolates of the CR1 community were identified as members of the genera Cellulomonas, Bacillus, Dietzia, and Halomonas. In light of our investigations, the bioremediation of contaminated saline soils should be quite possible if the salinity of the soil water is lower than 15% or if it is reduced below this limit by the addition of water. PMID- 14735222 TI - Expression and characterization of the 42 kDa chitinase of the biocontrol fungus Metarhizium anisopliae in Escherichia coli. AB - Albeit Metarhizium anisopliae is the best-characterized entomopathogenic fungus, the role of some hydrolytic enzymes during host cuticle penetration has not yet been established. Three chitinase genes (chit1, chi2, chi3) from Metarhizium have already been isolated. To characterize the chitinase coded by the chit1 gene, we expressed the active protein (CHIT42) in Escherichia coli using a T7-based promoter expression vector. The recombinant protein, CHIT42, is active against glycol chitin and synthetic N-acetylglucosamine (GlcNAc) dimer and tetramer substrates. These activities suggest that the recombinant CHIT42 acts as an endochitinase. PMID- 14735223 TI - FeoB is not required for ferrous iron uptake in Campylobacter jejuni. AB - Among strains of Campylobacter jejuni, levels of ferrous iron (Fe2+) uptake was comparable. However, C. jejuni showed a lower level of ferrous iron uptake than Escherichia coli. Consistent with studies of E. coli, Fe2+ uptake in C. jejuni was significantly enhanced by low Mg2+ concentration. The C. jejuni genome sequence contains a single known ferrous iron uptake gene, feoB, whose product shares 50% amino acid identity to Helicobacter pylori FeoB and 29% identity to E. coli FeoB. However, Fe2+ uptake could not be attributed to FeoB for several reasons. Site-directed mutations in feoB caused no defect in 55Fe2+ uptake. Among C. jejuni strains, various nucleotide alterations were found in feoB, indicating that some C. jejuni feoB genes are defective. In addition, uptake could not be attributed to the magnesium transporter CorA, since no reduction in 55Fe2+ uptake was observed in the presence of a CorA-specific inhibitor. PMID- 14735224 TI - Upregulation of Fas and FasL expression in testosterone-induced apoptosis of macrophages. AB - To the best of the authors' knowledge, there have been few reports on the effect of testosterone on the apoptosis of macrophages. In this report, we studied the effect of testosterone on the apoptosis of bone marrow-derived macrophages (BMMs) and the function of the Fas/Fas ligand (FasL) system in the process. Results showed that testosterone treatment in vitro at the physiological concentration of 10 nM did not induce the apoptosis of BMMs. However, BMMs underwent apoptosis when treated at higher concentrations of testosterone (100, 200 and 400 nM). Testosterone-induced apoptosis was associated with the enhanced expression of Fas, FasL, and caspase-8. These data suggest that the Fas/FasL system may play an important role in the testosterone-induced apoptosis of macrophages. PMID- 14735225 TI - Further evidence for suicide inhibition of semicarbazide-sensitive amine oxidase in guinea pig lung by 2-bromoethylamine. AB - The inhibitory effects of 2-bromoethylamine (2-BEA), a derivative of ethylamine, on guinea pig lung semicarbazide-sensitive amine oxidase (SAO) have been studied. Preincubation with 2-BEA time-dependently inhibited SSAO activity. The mode of the initial phase of inhibition was competitive, with a Ki value of 52 microM. After preincubation at 37 degrees C for 2 h, the inhibition was noncompetitive and irreversible, as there was no recovery of SSAO activity by dilution of the inhibited samples. Kinetic analyses confirmed previous results with rat lung SSAO that 2-BEA is a suicide SSAO inactivator with a dissociation constant of 42 microM. This latter value is similar to that of the Ki value (52 microM) for the reversible phase of inhibition by 2-BEA. Addition of the nucleophilic compound 2 mercaptoethanol could not reduce the SSAO inhibition, indicating that inactivation could not be prevented by trapping the enzymatic reaction product from 2-BEA. This finding clearly indicates that the reaction product should not diffuse away from its site of genesis and agrees with one of the characteristics of suicide inhibitors. This conclusively excludes the possibility of an affinity labeling mechanism. PMID- 14735226 TI - Involvement of adenosine A1 receptors in forced walking stress-induced analgesia in mice. AB - We investigated the involvement of adenosine receptors on forced walking stress induced analgesia using a formalin-induced paw-licking test in male mice. Exposure to forced walking stress for 6 h showed stress-induced analgesia in the second phase (10-30 min), but not in the first phase (0-10 min). In the second phase, forced walking stress-induced analgesia was blocked by theophylline, a nonselective adenosine-receptor antagonist and DPCPX, an adenosine A1-receptor antagonist, but not ZM 241385, an adenosine A2A-receptor antagonist. These findings suggest that adenosine A1 receptors are involved in the analgesic mechanism activated by the forced walking stress. PMID- 14735227 TI - Effects of propranolol on xenobiotic enzyme activities in rat type II pneumocytes and alveolar macrophages in vivo. AB - The accumulation of basic drugs (cationic amphiphilic), such as beta-adrenergic antagonists, by pulmonary tissue is well known. Ring hydroxylation of nonselective beta-adrenergic blocking agent propranolol is mediated mainly by cytochrome P450 (CYP) 2D6 and N-desisopropylation by CYP1A2 in human and rat liver microsomes. In this study, the repeated administration of propranolol resulted in a marked inhibition of hepatic metabolism and an increase in its systemic availability, due to covalent binding of reactive metabolites (formed from 4-OH-propranolol) to liver microsomal P4502D enzymes. The absence of CYP1A2 and the presence of CYP2D in the lung suggest a different pulmonary metabolism of propranolol in comparison with those in the liver. In this study, we investigated its effects in vivo on some xenobiotic-metabolizing enzymes in rat type II pneumocytes (RTII) and rat alveolar macrophages (RAM). Twenty hours after the last multiple (7 days) oral administration, propranolol (100 mg/kg b.w.) decreased NADPH cytochrome c reductase activity and cytochrome P-450-dependent dealkylation of 7-benzyloxyresorufin (BROD) (CYP1A1, 2A1, 3A1) and 7 ethoxyresorufin (EROD) (CYP1A1) in RTII, while glutathione-S-transferase (GST), DT-diaphorase (QR), gamma-glutamyl transferase (gamma-GT) activities, intracellular reduced glutathione level and dealkylation of 7-pentoxyresorufin (PROD) (CYP2B1) were not changed. It was found that propranolol significantly increased NADPH cytochrome c reductase and BROD activities in RAM. The results suggest a different susceptibility of RTII and RAM to propranolol and its contrary effects on lung xenobiotic-metabolizing enzyme activities in both types of cells. PMID- 14735228 TI - Attenuation of renal ischemia-reperfusion injury by trimetazidine: evidence of an in vivo antioxidant effect. AB - Renal ischemia-reperfusion injury constitutes the most common pathogenic factor for acute renal failure and is the main contributor to renal dysfunction in allograft recipients and revascularization surgeries. Many studies have demonstrated that reactive oxygen species play an important role in ischemic acute renal failure. The aim of the present study was to investigate the effects of the synthetic antiischemic agent trimetazidine in a rat model of renal ischemia-reperfusion injury. Renal ischemia-reperfusion was induced by clamping the unilateral renal artery for 45 min followed by 24 h of reperfusion. Trimetazidine (2.5 mg/kg i.p.) was administered 24 and 12 h prior to renal artery occlusion and the same dose was given intravenously 1 h before inducing ischemia. Tissue lipid peroxidation was measured as thiobarbituric acid reacting substances (TBARS) in kidney homogenates. Renal function was assessed by estimating serum creatinine, blood urea nitrogen (BUN), creatinine and urea clearance. Renal morphological alterations were assessed by histopathological examination of hematoxylin-eosin stained sections of the kidneys. Ischemia-reperfusion produced elevated levels of TBARS and deteriorated the renal function as assessed by increased serum creatinine, BUN and decreased creatinine and urea clearance compared with sham operated rats. The ischemic kidneys of rats showed severe hyaline casts, epithelial swelling, proteinaceous debris, tubular necrosis, medullary congestion and hemorrhage. Trimetazidine markedly reduced elevated levels of TBARS and significantly attenuated renal dysfunction and morphological changes in rats subjected to renal ischemia-reperfusion. These results clearly demonstrate the in vivo antioxidant effect and the therapeutic potential of trimetazidine, an anti-ischemic agent, in attenuating renal ischemia-reperfusion injury. PMID- 14735229 TI - Effects of the antidepressant drug moclobemide on learning and memory in rats. AB - Moclobemide is a well known drug with antidepressant action. The aim of this study was to investigate the effects of moclobemide on learning and memory processes in Sprague Dawley rats. Over a 5-day period, learning sessions with 30 trials per day and memory retention tests were performed. The conditioned responses (avoidances), the unconditioned responses (escapes) and the intertrial crossings were observed. An active avoidance test was carried out using a shuttle box. Two passive avoidance tests were used: step-through (using a light chamber) and step-down (using a platform). In the step-through passive avoidance test, the learning and retention sessions consisted of three trials each and the latency of reaction times (the rat remaining in the light chamber for more than 180 sec) was used as criterion. In the step-down passive avoidance test, learning and retention sessions consisted of two trials and the latency of reaction times (the rat remaining on the platform for 60 sec) was used as criterion. In the active avoidance tests, moclobemide dose-dependently increased the number of avoidances during learning sessions and maintained this number in memory retention tests. Moclobemide did not alter the number of escapes, but did increase motor activity. In the passive avoidance tests, moclobemide also increased the latency of reaction times in learning and short memory retrieval tests. These findings suggest that moclobemide improves learning and memory processes in active and passive avoidance tests and has a cognition-enhancing effect. PMID- 14735231 TI - Influence of an acidic beverage (Coca-Cola) on the pharmacokinetics of phenytoin in healthy rabbits. AB - This study was carried out to evaluate the influence of an acidic beverage (Coca Cola) on the pharmacokinetics of phenytoin in rabbits. In a cross-over study, phenytoin was given orally at a dose of 30 mg/kg and blood samples were taken at different intervals from 0-24 h. After a washout period of 7 days, Coca-Cola (5 ml/kg) was administered in combination with phenytoin (30 mg/kg) and blood samples were taken at various time intervals from 0-24 h. The same rabbits continued to receive Coca-Cola (5 ml/kg) for another 7 days. On the 8th day, Coca Cola (5 ml/kg) in combination with phenytoin (30 mg/kg) was administered and blood samples were taken at similar intervals. Plasma was separated and assayed for phenytoin by high performance liquid chromatography (HPLC) and various pharmacokinetic parameters were calculated. It was concluded that an acidic beverage (Coca-Cola) increases the extent of absorption of phenytoin by significantly increasing the Cmax and AUC(o-a) of phenytoin. These results warrant the reduction of phenytoin dose when administered in combination with Coca-Cola to avoid any toxicity. PMID- 14735230 TI - Endothelin-like action of Pausinystalia yohimbe aqueous extract on vascular and renal regional hemodynamics in Sprague Dawley rats. AB - The bark of the African tree Pausinystalia yohimbe has been used as a food additive with aphrodisiac and penile erection enhancing properties. The effect of an aqueous extract of P. yohimbe (CCD-X) on renal circulation was assessed in order to test the hypothesis that it possesses additional effects on nitric oxide production and/or endothelin-1 (ET-1)-like actions. In vivo studies with CCD-X in Sprague Dawley rats demonstrated a dose-dependent (1-1000 ng/kg) increase in mean blood pressure (p < 0.001) and an increase in medullary blood flow (MBF) (p < 0.001). Both the pressor action and renal medullary vasodilation were blocked by endothelinA (ETA) receptor antagonist BMS182874 and endothelinB (ETB) receptor antagonist BQ788 in combination. L-Nomega-nitro-l-arginine methyl ester (L-NAME; 10 mg/kg) also inhibited the increase in MBF induced by CCD-X. In vitro studies in isolated perfused kidney and in pressurized renal microvessels confirmed the dose-dependent vasoconstrictor action of this extract. ETA receptor antagonist BQ610 and ETB receptor antagonist BQ788 separately and significantly attenuated the renal vasoconstrictor actions of the extract (p < 0.001 ANOVA). These preliminary observations indicate that, in addition to the alpha-adrenergic antagonist actions that characterize yohimbine, CCD-X possesses endothelin-like actions and affects nitric oxide (NO) production in renal circulation. These findings suggest a strong possibility of post-receptor cross-talk between alpha2 adrenoceptors and endothelin, as well as a direct effect of alpha2-adrenoceptors on renal NO production. PMID- 14735232 TI - Snapshots in cardiology--new trials in cardiovascular pharmacotherapy: a report from the American Heart Association 2003 Scientific Sessions. PMID- 14735233 TI - Gateways to clinical trials. AB - Gateways to Clinical Trials is a guide to the most recent clinical trials in current literature and congresses. The data in the following tables has been retrieved from the Clinical Studies Knowledge Area of Prous Science Integrity, the drug discovery and development portal, http://integrity.prous.com. This issue focuses on the following selection of drugs: Abetimus sodium, adalimumab, alefacept, alemtuzumab, almotriptan, AMGN-0007, anakinra, anti-CTLA-4 Mab, L arginine hydrochloride, arzoxifene hydrochloride, astemizole, atazanavir sulfate, atlizumab; Belimumab, BG-9928, binodenoson, bosentan, botulinum toxin type B, bovine lactoferrin, BufferGel; Caspofungin acetate, ciclesonide,cilomilast, ciluprevir, clofarabine, CVT-3146; Darbepoetin alfa, desloratadine, diflomotecan, doripenem, dronedarone hydrochloride, drotrecogin alfa (activated), DT388-GM-CSF, duloxetine hydrochloride, E-5564, efalizumab, enfuvirtide, esomeprazole magnesium, estradiol acetate, ETC-642, exenatide, exisulind, ezetimib; Febuxostat; Gallium maltolate, ganirelix acetate, garenoxacin mesilate, gefitinib; H11, HuMax; IL-15, IDD-1, IGIV-C, imatinib mesylate, ISIS-14803, ITF 1697, ivabradine hydrochloride; KRN-5500; L-365260, levetiracetam, levosimendan, licofelone, linezolid, LJP-1082, lopinavir lumiracoxib; MCC-478, melatonin, morphine hydrochloride, morphine-6-glucuronide, moxidectin; N-Acetylcarnosine, natalizumab, NM-702, NNC-05-1869, NSC-703940; Ocinaplon OM-89, omalizumab, omeprazole/ sodium bicarbonate, OPC-28326, ospemifene; PEG-filgrastim peginterferon alfa-2a, pegsunercept, pirfenidone, pralmorelin, pregabalin; Recombinant glucagon-like peptide-1 (7-36) amide, repifermin, RSD-1235; S-8184, selodenoson, sodium dichloroacetate, suberanilohydroxamic acid; TAS-102, terfenadine, teriparatide, tipranavir troxacitabine; Ximelagatran; YM-337. PMID- 14735234 TI - Preliminary in vivo pharmacokinetic and pharmacodynamic evaluation of a novel calcineurin-independent inhibitor of NFAT. AB - A-285222 (A-285) is a bis-trifluoromethyl-pyrazole (BTP), a novel class of immunosuppressive agents that inhibit NFAT activity in vitro in human and non human primate cells through a calcineurin-independent mechanism. In this preliminary study, we treated cynomolgus monkeys with different doses of A-285 for several days. Blood was collected from all animals at different times during the study. From these samples, plasma concentrations of A-285 were measured by liquid chromatography/mass spectrometry (LC/MS), and intracellular T-cell production of the cytokines IL-2, IFN-gamma, and TNF-alpha was quantified by flow cytometry using a mitogen-stimulated whole blood assay. Marked inhibition of cytokine production occurred after administration of the first dose of A-285, and this effect was comparable to that of cyclosporine. While neurological toxic side effects were seen when the plasma concentration of A-285 exceeded 4 microg/ml, at lower plasma levels the drug was well tolerated over 2 weeks and its pharmacodynamic effects were sustained throughout this time. PMID- 14735235 TI - Can the experienced ICU physician predict ICU length of stay and outcome better than less experienced colleagues? AB - OBJECTIVE: To assess the ability of physicians with varying degrees of experience to predict the length of stay and outcome of intensive care unit (ICU) patients. DESIGN: Prospective, interview-based study. SETTING: A 31-bed mixed medical surgical ICU. PATIENTS: A total of 223 consecutive patients (excluding those admitted for routine post-operative surveillance) admitted to the ICU. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Physicians immediately responsible for each patient, and others fully aware of the case, were interviewed separately during the first 12 h of ICU admission to determine their assessment of the patient's likely duration of stay on the ICU and the probable outcome. Degree of predictive accuracy was assessed using the Kappa statistic with kappa < or =0.2 poor, 0.21-0.4 fair, 0.41-0.60 moderate, 0.61-0.8 good, and 0.81-1.0 very good. Physicians were graded according to their degree of experience as junior (less than 1 year ICU experience), medium (critical care fellow), and senior (staff physician with supervising functions). For lengths of stay less than 5 days, senior physicians were better predictors than less experienced doctors. For outcome prediction, physicians were generally moderately good at predicting death, with senior physicians tending to be more accurate than their less experienced colleagues (senior kappa 0.68, medium kappa 0.52, junior kappa 0.43). CONCLUSIONS: Prediction of length of ICU stay was poor amongst all physicians in patients with a length of stay greater than 5 days. Experienced physicians were better predictors of ICU lengths of stay less than 5 days and, in contrast to some reports, of ICU outcome than their more inexperienced counterparts. PMID- 14735236 TI - Albumin dialysis: a new therapeutic strategy for intoxication from protein-bound drugs. AB - OBJECTIVE: Although water-soluble drugs can be removed by haemofiltration/haemodialysis, morbidity and mortality from intoxication with protein-bound drugs remains high. The present study investigates whether albumin dialysis in the form of the Molecular Adsorbents Recirculating System (MARS) is effective in removal of protein-bound drugs. DESIGN: Prospective animal study. SETTING: Surgical research laboratory in a university hospital. SUBJECTS: Seven female Norwegian Landrace pigs. INTERVENTION: We studied whether midazolam (97% albumin-bound) and fentanyl (85% alpha-1-acid glycoprotein-bound), administered as anaesthetics to pigs with induced acute liver failure, could be removed by MARS dialysis lasting for 4 h. MEASUREMENTS: After 4 h of dialysis, total and free anaesthetic concentrations were measured in the blood and dialysate from different segments of the MARS circuit. MAIN RESULTS: Midazolam: total plasma concentrations fell by 47.1+/-2.1% (in 4 h) across the MARS filter ( p<0.01). The charcoal component of the system reduced the total dialysate drug concentration by 16.4+/-2.2% ( p<0.05). Free midazolam removal followed a similar pattern. Fentanyl: total plasma concentrations fell by 56.1+/-2.4% (in 4 h) across the MARS filter ( p<0.01). Clearance of fentanyl from the dialysate by the charcoal was 70+/-0.7% at 4 h ( p<0.001). CONCLUSIONS: The results of the study show that MARS can remove both albumin and other protein-bound drugs efficiently from the plasma, and it may have a place for the treatment of patients suffering from intoxication with this class of compounds. PMID- 14735237 TI - Non-invasive negative and positive pressure ventilation in the treatment of acute on chronic respiratory failure. AB - OBJECTIVE: To investigate in clinical practice the role of non-invasive mechanical ventilation in the treatment of acute respiratory failure on chronic respiratory disorders. DESIGN: An 18 months prospective cohort study. SETTING: A specialised respiratory intensive care unit in a university-affiliated hospital. PATIENTS: A total of 258 consecutive patients with acute respiratory failure on chronic respiratory disorders. INTERVENTIONS: Criteria for starting non-invasive mechanical ventilation and for endotracheal intubation were predefined. Non invasive mechanical ventilation was provided by positive pressure (NPPV) ventilators or iron lung (NPV). RESULTS: The main characteristics of patients (70% with chronic obstructive pulmonary disease) on admission were (mean, SD or median, 25th-75th centiles): pH 7.29 (0.07), PaCO(2) 83 mm Hg (19), PaO(2)/FiO(2) 198 (77), APACHE II score 19 (15-24). Among the 258 patients, 200 (77%) were treated exclusively with non-invasive mechanical ventilation (40% with NPV, 23% with NPPV, and 14% with the sequential use of both), and 35 (14%) with invasive mechanical ventilation. In patients in whom NPV or NPPV failed, the sequential use of the alternative non-invasive ventilatory technique allowed a significant reduction in the failure of non-invasive mechanical ventilation (from 23.4 to 8.8%, p=0.002, and from 25.3 to 5%, p=0.0001, respectively). In patients as a whole, the hospital mortality (21%) was lower than that estimated by APACHE II score (28%). CONCLUSIONS: Using NPV and NPPV it was possible in clinical practice to avoid endotracheal intubation in the large majority of unselected patients with acute respiratory failure on chronic respiratory disorders needing ventilatory support. The sequential use of both modalities may increase further the effectiveness of non-invasive mechanical ventilation. PMID- 14735238 TI - [Diagnosis and therapy of biliary tract diseases. An assessment of the current status]. PMID- 14735239 TI - [Gallbladder calculi--always an indication for surgery?]. AB - There is consensus that symptomatic cholecystolithiasis presents an indication for cholecystectomy. Today the surgical method of choice is the laparoscopic technique, which has proven its superiority in numerous randomized studies. Epidemiologic studies showed that 10-15% of all gallstone patients develop complications so that a prophylactic cholecystectomy is repeatedly being discussed. A few older studies based on conventional cholecystectomies, however, showed no decisive advantage for a prophylactic cholecystectomy, but rather clearly higher costs. Therefore a wait-and-see policy is generally recommended for asymptomatic gallstones. The analysis of our large group of patients showed that acute cholecystitis as well as common bile duct stones occur significantly more often with increasing duration of the gallstone disease. The older the patient, the longer the operation time, the more frequent a conversion, and the higher the morbidity of the procedure. Considering the minor impairment of daily activities with the laparoscopic technique, the present concept of treatment for asymptomatic and mildly symptomatic cholecystolithiasis needs to be scrutinized. PMID- 14735240 TI - [Primary biliary liver cirrhosis and overlap syndrome. Diagnosis and therapy]. AB - Primary biliary cirrhosis represents a chronic cholestatic liver disease of unknown etiology. It primarily affects females, is associated with extrahepatic immune-mediated syndromes, shows an immunogenetic association with HLA DR8, and displays serum autoantibodies, which makes an autoimmune etiology likely. The diagnosis is reached in patients with elevated alkaline phosphatase, gamma glutamyl transferase and bilirubin levels who exhibit normal bile ducts upon ultrasound examination, and in whom specific antimitochondrial autoantibodies are detectable. Half of all PBC patients additionally show specific antinuclear autoantibodies. Immunosuppressive therapy is ineffective; steroids, transplant immunosuppressants, colchicine, d-penicillamine and methotrexate are of limited clinical benefit. Ursodeoxycholic acid has few side effects and leads to a biochemical response and a delay of disease progression in most cases. When ursodeoxycholic acid therapy is ineffective an overlap syndrome with autoimmune hepatitis can be present, which can respond to steroid treatment. The only curative option is liver transplantation which should be considered when bilirubin levels exceed 100 microM/l. PMID- 14735241 TI - [Primary sclerosing cholangitis]. AB - Primary sclerosing cholangitis (PSC) is a chronic cholestatic liver disease, characterized by fibrosing inflammation and obliteration of intra and/or extrahepatic bile ducts. The disease belongs to the most common cholestatic diseases in adults and at present is diagnosed with increasing frequency. It is very often associated with ulcerative colitis. Patients with PSC have an increased incidence of bile duct carcinomas and those with ulcerative colitis also have an increased incidence of colonic carcinomas. Immunosuppressive treatment is little effective. Ursodeoxycholic acid (UDCA) has been shown to improve liver histology in PSC. The aim is to treat patients as early as possible to prevent progression to the advanced stages of the disease. During treatment with UDCA stenoses of major ducts may develop and early endoscopic dilatation is highly effective. In patients with endstage disease, UDCA is not effective and liver transplantation is indicated. PMID- 14735242 TI - [Gallbladder and bile duct carcinoma. Biology and pathology]. AB - Biliary tract cancers are a consequence of a stepwise malignant transformation of the biliary epithelium. Intrahepatic cholangiocarcinomas arises from any portions of the intrahepatic bile duct epithelium: the segmental or area ducts and their finer branches or intrahepatic small bile ducts. Cholangiocarcinoma arising from the hepatic ducts or near their junction are called hilar cholangiocarcinoma or Klatskin tumour, and are considered as extrahepatic lesion. Cancer of extrahepatic bile ducts may also arise in the Ductus cysticus or choledochus as well as in the gall bladder. For intrahepatic cholangiocarcinoma, the UICC-TNM classification system of malignant liver tumors is applied, differing from the UICC-TNM staging system of extrahepatic bile ducts and gall bladder carcinomas. The cause of carcinomas of the bile ducts remains speculative in most cases. However, chronic inflammation due to sclerosing cholangitis, hepatolithiasis or parasites is associated with carcinogenesis. Histopathologically, the vast majority are adenocarcinomas; mesenchymal tumors and primary melanomas are extremely rare. Different genetic alterations are discussed to be of importance. PMID- 14735243 TI - [Diagnosis of biliary tract carcinoma]. AB - There are several diagnostic tools available in the diagnosis of bile duct cancer. Tumors of the middle and distal part of the extrahepatic bile duct are accessible to endosonography. Endoscopic retrograde cholangiopancreatography (ERC) and percutaneous transhepatic cholangiography (PTC) are the most invasive procedures for diagnosis of bile duct cancer. However, they offer the opportunity to obtain material for cytological or histological investigation. Moreover, bile flow can be assured by inserting endoprostheses during the procedure. Cholangioscopy and/or intraductal ultrasonography can be performed during ERC. They confer to the diagnosis of a malignant bile duct tumor and are the most accurate methods to diagnose the extent of longitudinal spread. Magnetic resonance imaging-cholangiography is an efficient diagnostic procedure which should be used first, if the bile duct tumor is located in the hilar region. PMID- 14735244 TI - [Conservative and interventional endoscopic therapy of biliary tract carcinoma]. AB - Endoscopic therapy is central to the palliative treatment of bile duct carcinoma. In obstructive jaundice, biliary drainage has few complications and relieves symptoms reliably. It can prevent further complications and is indispensable to the treatment of cholangitis. The principal drawback of biliary stents is stent occlusion and cholangitis. Prophylactic antibiotics were not proven to be effective. Technical details concerning material, number and location of stents await further clarification. Photodynamic therapy is an emerging adjunct to palliative therapy of biliary cancer. Preoperative biliary drainage in obstructive jaundice is not warranted as a routine intervention. It may be indicated, though, as preoperative bridging or to allow liver function to recover before major hepatic surgery. Finally, stenting of postoperative bile duct stenosis is gaining increasing acceptance. PMID- 14735245 TI - [Generalized pain syndrome, fever and somnolence in an 81-year-old patient]. AB - We present a case of an 81-year-old diabetic man with anaerobic sepsis due to acalculous cholecystitis. The patient was admitted to our hospital with a seven day history of severe abdominal pain accompanied with fever and somnolence. Blood cultures taken during the initial procedure developed Clostridium perfringens. The patient was immediately treated with parenteral penicillin. The ultrasonography pointed out the case: the gall bladder was found to be distended and slightly thickened. This result was interpreted as an acute non-emphysematous cholecystitis. The material obtained by needle aspiration and therapeutical emptying of the gall bladder revealed large gram positive rods, that also proved to be Clostridium perfringens. The patients course afterwards was uneventful. Antibiotics were continued and he was discharged after 13 days in a stable condition. PMID- 14735246 TI - [Lymphadenopathy in the groin. Unusual etiology in a 65-year-old patient]. AB - A 65 year old female patient was admitted to our hospital with unspecific symptoms and a lump in the right groin which raised suspicion of a malignant lymphoma. Histologically a follicular dendritic reticular cell tumor was found. Because there are aggressive forms of this tumor and no established standard therapy, we decided to treat her with surgery followed by a combined radio- and chemotherapy. Despite this treatment 16 months after the first diagnosis a relapse occurred with a metastasis of the follicular dendritic cell tumor in the lung. The metastasis was resected surgically. The optimal management of this kind of tumor is not known. PMID- 14735247 TI - [Modern pharmacotherapy of Hodgkin disease]. AB - High cure rates have been achieved in the treatment for patients with Hodgkin's disease in the past 30 years. Depending on stage at diagnosis and further risk factors up to 95% of patients with Hodgkin's disease can be cured with first-line treatment. Modern therapeutic strategies aim at both reducing therapy-induced late toxicities while maintaining effective tumor control. Patients with early stage Hodgkin's disease are now treated with a short course of chemotherapy for control of occult disease and involved field (IF) irradiation. For patients with early unfavourable stages, effectiveness of treatment shall be optimised by introducing the escalated BEACOPP schedule which has been established in the treatment of advanced stages. Questions to be answered in the treatment of advanced stages concern the optimal number of cycles of an effective chemotherapy regimen and the necessity of additional radiation therapy. The role of erythropoetin and PET-imaging is currently being evaluated in ongoing trials. In the future, new therapeutic approaches with biological agents will be of interest. PMID- 14735248 TI - [Imatinib-New, molecular therapeutic possibility in chronic myeloid leukemia. The IRIS Study]. PMID- 14735249 TI - [New studies in cardiology. Report of the European Cardiology Congress 30 August 3 September 2003 in Vienna]. PMID- 14735250 TI - An equilibrium thermodynamic model of the sequestration of calcium phosphate by casein phosphopeptides. AB - Sequestration of calcium phosphate by caseins occurs in the Golgi region of mammary secretory cells during lactation, where it helps to prevent calcification of the gland and to deliver high concentrations of calcium and phosphate to the neonate in the form of milk. Calcium phosphate nanoclusters are formed when a core of amorphous calcium phosphate is sequestered within a shell of casein or casein phosphopeptides. The nanoclusters can form spontaneously from a supersaturated solution or by dispersion of a precipitate of calcium phosphate, demonstrating that they are thermodynamically stable complexes. The average size and chemical composition of the complexes are largely independent of the solution conditions (pH, temperature, peptide concentration, salt composition and rate of reaction) under which they form. Larger, metastable, colloidal particles can form if there is not enough of the phosphopeptide to sequester all the calcium phosphate, or, transiently, if the salt and peptide solutions are mixed together without sufficient care. A thermodynamic model of the sequestration process is presented which makes use of an invariant ion activity product observed in nanocluster-containing solutions. In any given solution that has thermodynamic stability, the extent of the sequestration reaction can be calculated from the empirical formula of the nanoclusters using the criterion that the solution should have the equilibrium value of the invariant ion activity product. Other members of the paralogous group of secretory calcium-binding phosphoproteins to which caseins belong may also be able to sequester calcium phosphate in biological fluids such as saliva and in the extracellular matrix of mineralizing tissues. PMID- 14735251 TI - An equilibrium thermodynamic model of the sequestration of calcium phosphate by casein micelles and its application to the calculation of the partition of salts in milk. AB - An equilibrium thermodynamic model of the interaction of calcium, phosphate and casein in milk is described in which the micellar calcium phosphate is assumed to be in the form of calcium phosphate nanoclusters. A generalized empirical formula for the nanocluster is used to define the molar ratios of small ions (Ca, Mg, P(i) and citrate) to a casein phosphorylated sequence (phosphate centre, PC). From this model, a method of calculating the partition of milk salts into diffusible and non-diffusible fractions is obtained. No arbitrary assumptions are made, no fitting of adjustable parameters is done and the PCs in the caseins are defined by inspection of their primary structures. In addition to the salt partition, the mole fractions of the individual caseins not complexed to the calcium phosphate through one or more of their PCs are computed and a generic stability rule for milks is derived. The use of the model is illustrated by calculations of the partition of salts in a standard milk and by comparison with experimental data on the partition of salts in the milk of individual cows. The generic stability rule is applied to the individual milks to determine whether the micellar calcium phosphate is thermodynamically stable. According to the calculations, compositions that might lead to pathological calcification in the lumen of the mammary gland were seldom found in primiparous healthy cows in early or mid lactation but occurred more often in multiparous animals, in late lactation and during mastitic infection. PMID- 14735252 TI - B-type natriuretic peptide levels in congenital heart disease. AB - The objective of this study was to evaluate the potential role of B-type natriuretic peptide (BNP) levels in children with congenital heart disease undergoing cardiac catheterization. Measurement of plasma BNP concentration has been shown to be useful in the diagnosis, risk stratification, and management of adult patients with congestive heart failure, but little is known about the role of BNP in children with structural congenital heart disease. We measured plasma BNP levels using the Triage BNP test in patients with congenital heart disease referred for diagnostic or interventional cardiac catheterization. Plasma BNP concentration was measured in 96 children and 11 adults > or = 19 years old (7.9+/-8.3 years) undergoing heart catheterization for underlying congenital heart disease. BNP levels ranged from < 5 to > 1300 pg/ml, with a median BNP concentration of 19.0 pg/ml. Baseline BNP concentrations were > 100.0 pg/ml on 19 occasions in 17 patients. The pressure difference between the left ventricle and ascending aorta was 10-110 mmHg in 21 patients. BNP concentrations for this cohort ranged from < 5.0 to 1060.0 pg/ml and correlated with the degree of left ventricular outflow obstruction (correlation coefficient, 0.661; p = 0.001). This study suggests that with additional research, BNP concentration may prove to be a useful clinical tool in managing children and adults with congenital heart disease. PMID- 14735253 TI - Repolarization abnormalities in children with a structurally normal heart and ventricular ectopy. AB - In adults, increased QT dispersion has been shown to predict arrhythmic risk as well as risk of sudden death in several clinical settings. It is controversial whether QT and JT dispersion are increased in children with ventricular ectopy and a structurally normal heart. We studied two groups of children: 25 patients with ventricular ectopy and 25 healthy children as controls. Standard electrocardiograms were reviewed and dispersions of both corrected QT (QTc) and JT (JTc) intervals were compared. We conclude that QTc and JTc dispersions are significantly increased in children with ventricular ectopy compared to control subjects. PMID- 14735254 TI - Alterations in the natriuretic hormone system related to cardiopulmonary bypass in infants with congestive heart failure. AB - This study examined changes in the natriuretic hormone system in five infants with congestive heart failure (CHF) due to intracardiac left-to-right shunting who were exposed to cardiopulmonary bypass (CPB) during surgical repair. Plasma concentrations of three hormones [atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP), and dendroaspis natriuretic peptide (DNP)] and their secondary messenger, guanosine 3',5'-monophosphate (cGMP), were measured, and the biological activity of the system was quantified. At baseline, BNP and DNP concentrations were normal in our patients, a finding that is strikingly different from that of adult CHF patients, whereas ANP concentrations were elevated. Following CPB, ANP concentrations decreased (median, 175 vs 44 pg/ml; p = 0.043) and BNP concentrations increased (median, 25 vs 66 pg/ ml; p = 0.043), whereas DNP concentrations did not change. Following modified ultrafiltration, BNP concentrations increased (p = 0.043), but other natriuretic peptide concentrations did not change. The calculated biological activity of the natriuretic hormone system decreased following CPB [molar ratio, cGMP / (ANP + BNP + DNP); median, 213 vs 127; p = 0.043)]. Additional studies are needed to expand on these findings and identify patients with other types of congenital heart disease who have perioperative disturbances in the natriuretic hormone system and thus might benefit from pharmacologic intervention. PMID- 14735257 TI - Early detection of drug interactions utilizing a computerized drug prescription handling system-focus on cerivastatin-gemfibrozil. AB - OBJECTIVES: Based on the recent cerivastatin experience, we retrospectively evaluated the effect of notifying a drug alert utilizing a computerized drug handling system. METHODS: The evaluation was carried out during three periods: period I corresponded to all prescriptions issued during April, 2001 ("baseline period"), before the Spanish Drug Agency issued alerts on the concomitant therapy with cerivastatin and gemfibrozil; period II (June) corresponded to a time in which a first informative note had been released; and period III (July) after the second warning alert was issued. RESULTS: Data collected included the reading of 2,693,656 drug prescriptions, 1,937,083 (71.9%) of which contained patient information. Forty-four patients received combined therapy with cerivastatin and gemfibrozil over the three periods, yielding 55 exposures: 27 during the baseline period, and 28 between periods II and III, when the alert bulletins had already been released. Moreover, 41.6% of doctors included in the follow-up repeated the hazardous prescription during those two periods. CONCLUSIONS: The effect of the informative notes about the risk of prescribing cerivastatin and gemfibrozil concomitantly on doctors' prescribing habits was limited. The system for screening information from drug prescriptions presented herein allows the early detection of drug interactions by identifying the doctors who issue hazardous prescriptions as well as patients at the highest risk of adverse drug reactions, thus allowing a personal feedback with both of them. PMID- 14735258 TI - Frequency and enzyme activity of the butyrylcholinesterase K-variant in a Turkish population. AB - OBJECTIVE: Among variants of the butyrylcholinesterase gene ( BChE), the K variant causing Ala539Thr substitution is the most common one associated with about one-third reduction in the enzyme activity. This study aimed to detect the frequency of the K-variant allele in a Turkish population sample and also to evaluate how the plasma BChE activity was influenced by this variant. METHODS: Patients administered for elective surgery ( n=77) were examined for the presence of the K allele. The enzyme activity was determined in plasma. RESULTS: The K variant of BChE is a common allele with a frequency of 0.266 (CI(95%) 0.196 0.336) in our sample from a Turkish population. Mean enzyme activity in subjects homozygous for the K-variant was about 40% lower than other subjects. CONCLUSION: The frequency of the BChE K-variant was significantly higher in a Turkish population than those reported for other populations and it is associated with a diminished enzyme activity. PMID- 14735261 TI - Acute effects of an oral calcium load on markers of bone metabolism during endurance cycling exercise in male athletes. AB - Although sport and physical activity are generally considered as positive factors for bone metabolism some endurance trainings such as running and bicycling have few or no beneficial or even deleterious effects on bone mineral density. The present study was designed to investigate the acute effect of an intensive endurance cycling exercise on biochemical bone markers. Furthermore, the effect of the oral intake of 1 g calcium load, by drinking high-calcium mineral water, just prior to and during the exercise was checked. Twelve well-trained elite male triathletes aged 23-37 years were explored. The serum concentrations of calcium, phosphate, PTH, bone alkaline phosphatase (BALP) and C-terminal cross-linking telopeptide of type 1 collagen (CTX) were measured before, during and after a 60 min 80% VO2max cycle ergometer exercise. Since cycling exercise was accompanied by a reduction in plasma volume the total amount of biochemical bone markers was calculated. When the exercise was performed without calcium load both serum concentrations and total amount of CTX began to increase progressively 30 min after the start of the exercise and were still significantly elevated, by 45-50%, 2h after the end of the exercise. Ingestion of high-calcium mineral water completely suppressed the CTX response. By contrast serum concentrations and total amount of BALP fluctuated and showed no significant difference with or without calcium load. The present study demonstrates that the burst of osteoclastic activity acutely induced by an endurance cycling exercise can be suppressed by the previous intake of a calcium load afforded by drinking high calcium mineral water. PMID- 14735268 TI - Speciation analysis of mercury in water samples by cold vapor atomic absorption spectrometry after preconcentration with dithizone immobilized on microcrystalline naphthalene. AB - Trace amounts of inorganic mercury (Hg(2+)) and methylmercury cations (MeHg(2+)) were adsorbed quantitatively from acidic aqueous solution onto a column packed with immobilized dithizone on microcrystalline naphthalene. The trapped mercury was eluted with 10 ml of 7 mol L(-1) hydrochloric acid solution. The Hg(2+) was then directly reduced with tin (II) chloride, and volatilized mercury was determined by cold vapor atomic absorption spectrometry (CVAAS). Total mercury (Hgt) was determined after decomposition of MeHg(+) into Hg(2+). Hg(2+) and MeHg(+) cations were completely recovered from the water with a preconcentration factor of 200. The relative standard deviation obtained for eight replicate determinations at a concentration of 0.3 microg L(-1 )was 1.8%. The procedure was applied to analysis of water samples, and the accuracy was assessed via recovery experiment. PMID- 14735270 TI - Production of certified reference materials for mycotoxins: IRMM's view on the assessment of uncertainties. AB - Analytical difficulty and the economic importance of controlling mycotoxin levels in food and feed led the Community Bureau of Reference (BCR) to prepare a series of certified reference materials (CRM) for various mycotoxins. Because of the wide acceptance of these CRM and the need to ensure the comparability and traceability of measurements in the future it is necessary to prepare and certify new batches of mycotoxin reference materials (RM). In the following text two different approaches for evaluation of the characterisation uncertainty of CRM will be compared using the certification of aflatoxin M(1) (AfM(1)) in milk powder as an example. The conventional approach is based on evaluation of characterisation exercise data; the alternative approach is based on measurement uncertainties of the employed analytical methods. Because laboratories are using totally different approaches to estimate the measurement uncertainties, combination of the uncertainties obtained from the participating laboratories was not recommended. Therefore, a new integrated approach for assessment of the measurement uncertainties of the analytical methods on the basis of additional data collected during the characterisation exercise will be described. The conventional approach was found to be the most appropriate and economical approach to evaluate the characterisation uncertainty as a characterisation exercise must be performed anyway to establish the property values of candidate (C)RM, irrespective of whether or not reliable measurement uncertainties can be provided by the laboratories. An integrated approach for assessment of measurement uncertainties based on additional characterisation data as applied here to enable use of an uncertainty-based approach provides more information but is too time-consuming and cost-intensive to become common practice. PMID- 14735271 TI - Application of the revised EU criteria for the confirmation of anabolic steroids in meat using GC-MS. AB - The EU criteria for the confirmation of the presence of illegal compounds in biological matrices were recently revised. The old and the revised criteria were applied to relative ion intensities obtained for five anabolic steroids (methylboldenone, methyltestosterone, ethynylestradiol, beta-boldenone and beta nortestosterone) in meat (cow, pig, turkey) and fish at concentrations ranging from 0.5 to 5.0 microg/kg. Confirmatory analysis was done by GC-MS; therefore four diagnostic ions had to be monitored and three ion ratios had to be calculated and tested against the criteria. Application of the old and revised criteria, with either standards or fortified samples as reference, showed mutually rather divergent results. Confirmation according to the revised EU criteria and using fortified samples as a reference gave the best results; in other words the highest percentage of diagnostic ion ratios within the tolerance intervals. A correlation was found between the percentage of these ion ratios and the signal/noise (S/N) ratio of the least intense ion of interest in the recorded MS spectrum. Although there were distinct differences in the results obtained for different analytes and sample types, it is safe to conclude that at S/N=3 the percentage of ratios within the tolerance intervals generally will be at or below 50%, while for S/N>/=10, the percentage increases to over 90%. In the present study, fully satisfactory results were obtained down to about 2 microg/kg, but not for lower analyte concentrations. PMID- 14735272 TI - Structural and optical characterization of pyrolytic carbon derived from novolac resin. AB - The structural and optical properties of technologically interesting pyrolytic carbons formed from cured novolac resin and cured novolac/biomass composites were studied by X-Ray Diffraction Analysis (XRD), and Fourier Transform Infrared (FTIR), Raman and Photoluminescence (PL) spectroscopy. Pyrolysis of the cured materials took place at temperatures in the range 400-1000 degrees C. The most important weight loss, shrinkage and structural changes of pyrolyzed composites are observed at temperatures up to 600 degrees C due to the olive stone component. In the same temperature range, the changes in pyrolyzed novolac are smaller. The spectroscopic analysis shows that novolac pyrolyzed up to 900 ( degrees )C has less defects and disorder than the composites. However, above 900 ( degrees )C, pyrolyzed novolac becomes more disordered compared to the pyrolyzed composites. It is concluded that partial replacement of novolac by olive stone in the composite materials leads to the formation of a low cost, good quality product. PMID- 14735273 TI - Thick-film electrodes for measurement of superoxide and hydrogen peroxide based on direct protein-electrode contacts. AB - Cytochrome c was immobilized on screen-printed thick-film gold electrodes by a self-assembly approach using mixed monolayers of mercaptoundecanoic acid and mercaptoundecanol. Cyclic voltammetry revealed quasi-reversible electrochemical behavior of the covalently fixed protein with a formal potential of +10 mV vs. Ag/AgCl. Polarized at +150 mV vs. Ag/AgCl the electrode was found to be sensitive to superoxide radicals in the range 300-1200 nmol L(-1). Compared with metal needle electrodes sensitivity and reproducibility could be improved and combined with the easiness of preparation. This allows the fabrication of disposable sensors for nanomolar superoxide concentrations. By changing the electrode potential the sensor can be switched from response to superoxide radicals to hydrogen peroxide-another reactive oxygen species. H(2)O(2) sensitivity can be provided in the range 10-1000 micromol L(-1) which makes the electrode suitable for oxidative stress studies. PMID- 14735274 TI - Improving tritium exposure reconstructions using accelerator mass spectrometry. AB - Direct measurement of tritium atoms by accelerator mass spectrometry (AMS) enables rapid low-activity tritium measurements from milligram-sized samples and permits greater ease of sample collection, faster throughput, and increased spatial and/or temporal resolution. Because existing methodologies for quantifying tritium have some significant limitations, the development of tritium AMS has allowed improvements in reconstructing tritium exposure concentrations from environmental measurements and provides an important additional tool in assessing the temporal and spatial distribution of chronic exposure. Tritium exposure reconstructions using AMS were previously demonstrated for a tree growing on known levels of tritiated water and for trees exposed to atmospheric releases of tritiated water vapor. In these analyses, tritium levels were measured from milligram-sized samples with sample preparation times of a few days. Hundreds of samples were analyzed within a few months of sample collection and resulted in the reconstruction of spatial and temporal exposure from tritium releases. Although the current quantification limit of tritium AMS is not adequate to determine natural environmental variations in tritium concentrations, it is expected to be sufficient for studies assessing possible health effects from chronic environmental tritium exposure. PMID- 14735275 TI - New NIST sediment SRM for inorganic analysis. AB - NIST maintains a portfolio of more than 1300 standard reference materials (SRM), more than a third of these relating to measurements in the biological and environmental fields. As part of the continuous renewal and replacement efforts, a set of new marine sediments has been recently developed covering organic and inorganic determinations. This paper describes the steps taken in sample preparation, homogeneity assay, and analytical characterization and certification with specific emphasis on SRM 2702 inorganics in marine sediment. Neutron activation analysis showed the SRM to be highly homogeneous, opening the possibility for use with solid sampling techniques. The certificate provides certified mass fraction values for 25 elements, reference values for eight elements, and information values for 11 elements, covering most of the priority pollutants with small uncertainties of only several percent relative. The values were obtained by combining results from different laboratories and techniques using a Bayesian statistical model. An intercomparison carried out in field laboratories with the material before certification illustrates a high commutability of this SRM. PMID- 14735277 TI - Structure elucidation of nanogram quantities of unknown designer drugs based on phenylalkylamine derivates by ion trap multiple mass spectrometry. AB - Multiple mass spectra (MS(1) to MS(6)) of 55 phenylalkylamine derivatives were recorded with ion-trap mass spectrometry employing electrospray (ESI) and atmospheric pressure chemical ionization (APCI). Fragmentation patterns were studied in detail and a generally applicable scheme was established for elucidation of the structures of phenylalkylamine derivatives. HPLC combined with ion-trap multiple mass spectrometry was used to identify the structure of reaction by-products in ecstasy samples from the "black" market. Low nanogram amounts were sufficient for on-line HPLC-MS(n) structure elucidation of unknowns. PMID- 14735278 TI - A method for near-infrared spectral calibration of complex plant samples with wavelet transform and elimination of uninformative variables. AB - An algorithm is proposed for extracting relevant information from near-infrared (NIR) spectra for multivariate calibration of routine components in complex plant samples. The algorithm is a combination of wavelet transform (WT) data compression and a procedure for uninformative variable elimination (UVE). After compression of the NIR spectra by WT, the UVE approach is used to eliminate the irrelevant wavelet coefficients. Finally, a calibration model is built from the retained wavelet coefficients to enable prediction. Because irrelevant information can be removed from the spectra used for multivariate calibration, the model based on the extracted relevant features is better than those obtained with full-spectrum data. Both prediction precision and calculation speed are improved. PMID- 14735279 TI - Direct optical resolution of the enantiomers of novel chiral tetrahedral metal clusters by HPLC on a cellulose tris-(3,5-dimethylphenylcarbamate) stationary phase. AB - The enantioseparation of seven novel chiral transition metal tetrahedral clusters has been achieved for the first time on cellulose derivatized with tris(3,5 dimethylphenylcarbamate) (CDMPC) as chiral stationary phase (CSP) and hexane containing different alcohols as modifiers as mobile phases. The effect of mobile phase composition on enantioselectivity was studied, and the effect of structural variation of the solutes on their enantioseparation was also investigated. It was found that both the metal in the tetrahedral core and the ligand coordinated to the atom in the tetrahedral core had significant effects on the chromatographic behavior of the analytes. PMID- 14735280 TI - Certified reference materials to support a European thematic strategy for soil protection: the EUROSOILS (IRMM-443) experience. AB - The recent proposal of thematic strategy for soil protection by the European Commission outlines that knowledge of problems associated with soil has increased considerably based on surveys, monitoring systems and data networks. Although this information is very useful, its value for the policy-making process is limited due to a lack of comparability. From this unsatisfying situation the need arises to develop an EU-wide monitoring system accompanied by suitable matrix reference materials. Experiences gained from the development and certification of IRMM-443 (The EUROSOILS) show how to approach the complex problem of providing suitable (i.e. representative) soil CRMs for the arising analytical needs to support existing and upcoming EU legislation. This paper presents briefly the quantitative findings of a certification exercise (adsorption coefficients for atrazine, lindane and 2,4-D, as well as pH in suspension). A comparison of these results with those of an intercomparison exercise organised in 1989 by the German UBA on very similar soils shows the significant improvement in the determination of soil adsorption coefficients that was induced by the EUROSOIL project. Indicative values of total and organic carbon content and nitrogen according to ISO Standards are also presented.Furthermore, the idea of reference soils and derived reference materials (for analytical purposes) as well as reference matrices (substrates for ecotoxicological testing) is highlighted and set into the context of horizontal standardisation of methods. PMID- 14735281 TI - Selenomethionine contents of NIST wheat reference materials. AB - Values of the total selenium and selenomethionine (Semet) content of four wheat based reference materials have been obtained by gas chromatography-stable isotope dilution mass spectrometry methods. The total Se method is an established one, and the results obtained with it are consistent with previously-assigned values. The Semet method (previously reported by our laboratory) is based on reaction with CNBr. Our data indicate that the four wheat samples (wheat gluten, durum wheat, hard red spring wheat, and soft winter wheat), though having a 30-fold range in total Se content, all have about 45% of their total Se values in the form of selenomethionine. Investigation of the CNBr-based method suggests that additional experiments are needed to verify that all selenomethionine in the wheat samples is accounted for, but also indicates that the values obtained are within 15% of the true values. As the form in which Se occurs in foods and dietary supplements is important from a nutritional perspective, adding information about Se speciation to total Se values in appropriate reference materials makes these materials more valuable in relevant analytical work. PMID- 14735282 TI - The need for reference materials when monitoring nitrate intake. AB - Whether dietary exposure to nitrate metabolites is detrimental or beneficial to human health has long been a matter of controversy. In spite of no consistent epidemiological evidence, nitrate metabolites are associated with the formation of carcinogenic-nitrosamines and gastric cancer. Furthermore, recent studies demonstrate that ingested nitrate plays a role in host defence against gastrointestinal pathogenic bacteria. Analytical values of nitrate content in foods are essential for estimating nitrate intake. The analytical process is of paramount importance for assessing human nitrate exposure and for establishing a link between these exposures and the current and future observed health effects. Therefore, the quality assurance of the measurement process is crucial to obtaining reliability, comparability and traceability of results. Certified Reference Materials (CRM) should play a role in the consistency of the measurement process. However, the availability of nitrate CRMs is still poor. When food monitoring is demanded, an approach could be to use In House Reference Materials (IHRM), prepared at a high metrological level, and all preparation steps should be quality driven. IHRMs were prepared, and available CRMs were used to provide traceability of the process. The homogeneity of IHRM was evaluated using an appropriate statistical design. The stability was monitored using an isochronous method. The material shelf life and storage conditions are presented. HPLC was optimised for the determination of nitrates in four vegetable categories. When a suite of IHRMs were used, the response of the HPLC system was linear over the range 1 to 8 mg L(-1). The detection limit for these compounds was 0.2 microg L(-1) and the determination limit 1.2 microg L(-1). The relationship between measurement uncertainty and critical points of the analytical process is presented. The differences in observed relative uncertainty between food categories could reflect current limitations in the food matrix reference materials. PMID- 14735283 TI - Production of a calibrant certified reference material for determination of the estrogenic mycotoxin zearalenone. AB - Several previous interlaboratory studies in the field of mycotoxin analysis have revealed considerable problems, apparent as high between-laboratory standard deviations, or rather non-comparable and non-traceable results. A major reason is lack of proper calibrants for external calibration. Public awareness of substances that mimic or interfere with the activity of natural hormones (endocrine disrupters) has led to increased interest in mycotoxins with estrogenic potential, e.g. zearalenone (ZON). During a large-scale standard measurement and testing (SMT) project of the European Commission (EC) dealing with the preparation and certification of reference materials for determination of the mycotoxin ZON in maize, a ZON calibrant in acetonitrile was prepared and intensively checked for purity, homogeneity, and stability. Preparation of the material, study of its homogeneity and stability, and characterisation of the calibrant on the basis of its preparation, with discussion of the results obtained, are described in this paper. The certified value of 9.95 micro g mL(-1) for ZON in acetonitrile and its corresponding expanded uncertainty of +/-0.30 micro g mL(-1) were calculated in compliance with the Guide to the Expression of Uncertainty in Measurement (GUM). PMID- 14735285 TI - ICP-MS trace-element analysis as a forensic tool. AB - Careful tracing of evidence at the site of a crime must be performed before suspects can be convicted of a crime or homicide. Fingerprints and ballistic control samples are important evidence. A common method used to examine lead bullets is comparison of physical properties such as weight, dimensions, shape, and distinctive markings. However ballistic investigations, for example comparison of characteristic scratches and marks left on fired bullets, do not always give sufficient information. Ballistic abrasion patterns can change for a variety of reasons, e.g. deformation or mechanical strain. Sometimes only particles remain in a victim's body. In such cases trace-element composition and lead-isotope ratios can be compared with those of controls. Elemental composition of particles and deformed bullets have been compared with the elemental fingerprints and isotope ratios of potential bullet types found on suspects. The applicability of the method was studied for two different cases. Data interpretation and several limitations of the technique are also discussed in this paper. PMID- 14735286 TI - Biomolecular and element analytical spectroscopy: new methods and application perspectives for two associated disciplines. PMID- 14735288 TI - Empathy and aggression: two faces of ecstasy? A study of interpretative cognitive bias and mood change in ecstasy users. AB - RATIONALE: As central 5-hydroxytryptamine (5-HT) is attenuated for a period following a single dose of MDMA ("ecstasy") and low 5-HT is associated with aggression, then MDMA users may be more aggressive in the days following an acute dose of the drug. OBJECTIVE: This study therefore aimed to determine if acute use of MDMA is associated with aggression 4 and 7 days later. METHODS: Twenty-nine MDMA users and 32 controls were compared on self-rated aggression and depression on the night of drug use (day 0), 4 and 7 days later. On day 4, participants performed an interpretative bias task in which they processed ambiguous sentences that could be interpreted in either an aggressive or neutral way (e.g. "The painter drew the knife"). RESULTS: MDMA users had faster response times in completing ambiguous aggressive sentences than neutral sentences; controls showed the opposite pattern of performance. In a subsequent recognition task, MDMA users were more confident in judging, and responded faster to, aggressive than neutral sentences; controls again showed the opposite pattern of effects. The level of aggressive interpretative bias positively correlated with extent of MDMA use. Midweek, MDMA users had higher self-rated aggression and depression scores than controls; on day 7, scores of both groups were similar. CONCLUSIONS: MDMA users display a cognitive bias towards interpreting ambiguous information in an aggressive way a few days after taking the drug. Self-rated mid-week low mood and mid-week aggression do not persist 7 days after use of the drug. This pattern of results is consistent both with the acute and residual effects of MDMA on central 5-HT and with the notion that 5-HT plays a role in modulating human aggression. PMID- 14735289 TI - The substantia nigra pars reticulata mediates the enhancement of startle by the dopamine D1 receptor agonist SKF 82958 in rats. AB - RATIONAL AND OBJECTIVES: Several studies have shown that the substantia nigra pars reticulata (SNr) is a critical site of action mediating dopamine agonist effects on motor behaviors. Because dopaminergic and GABA ergic mechanisms may interact in the SNr, we tested the contribution of both dopamine and GABA receptors in the SNr on the enhancement of startle by the dopamine D1 agonist SKF 82958. METHODS: Male Sprague-Dawley rats were implanted with cannulae into the SNr and 1 week later infused with either the D1 antagonist SCH 23390 (0.1, 1 microg) or the GABA(A) antagonist bicuculline (0.1 microg), followed by a systemic challenge with the D1 agonist SKF 82958 (1 mg/kg). Other rats were infused with the GABA(A) agonist muscimol (0.1 microg) or SKF 82958 (0.1, 1, 5 microg). RESULTS: Both SCH 23390 and bicuculline infused into the SNr completely blocked the enhancement of startle by systemic SKF 82958. Muscimol infused into the SNr produced a significant increase in startle by itself, whereas SKF 82958 had no effect. CONCLUSIONS: These results suggest that activation of D1 receptors in the SNr is necessary for the enhancement of startle by SKF 82958, but that activation of these receptors alone is not sufficient to increase startle. These results also suggest that GABA transmission in the SNr may be involved in the enhancement of startle by SKF 82958. Based on these data, we propose that activation of striatonigral neurons by D1 receptor agonists facilitates GABA release in the SNr to produce the observed enhancement of startle. PMID- 14735290 TI - Changes in extracellular 5-HIAA concentrations as measured by in vivo microdialysis technique in relation to changes in 5-HT release. AB - RATIONALE: The cerebral microdialysis technique has been widely used to monitor the release of 5-hydroxytryptamine (5-HT). The extracellular concentration of 5 HT has generally been shown to change after pharmacological manipulation as expected. Extracellular levels of the metabolite, 5-hydroxyindoleaceticacid (5 HIAA) does not always change in the same direction as 5-HT and has therefore generally been thought to be of no interest as a marker for 5-HT release. OBJECTIVE: The aim of the present review is to analyse the connection between changes in extracellular levels of 5-HT and 5-HIAA evoked by various pharmacological means. METHODS: Literature on in vivo microdialysis studies measuring extracellular 5-HT and 5-HIAA has been analysed with special attention to the great importance of the 5-HT re-uptake mechanism in determining their extracellular concentrations. RESULTS: When the 5-HT reuptake mechanism is intact changes in extracellular levels of 5-HT and 5-HIAA go in the same directions, e.g decrease after compounds that decrease 5-HT release and increase after compounds that enhance 5-HT release. Because the extracellular 5-HIAA concentrations is 100 1000 times higher than that of 5-HT similar percentage changes imply that a very small part of the released 5-HT reaches the microdialysis probe under these conditions. When the 5-HT reuptake mechanism is blocked the extracellular 5-HT increases whereas extracellular 5-HIAA decreases mainly because of the 5-HT(1B) receptor-induced decrease in 5-HT release but in part also because of the inhibition of reuptake of 5-HT, both resulting in decreased formation of 5-HIAA. CONCLUSION: Drug-induced changes in extracellular 5-HIAA levels can give valuable information on the effects of these drugs on the 5-HT release. PMID- 14735291 TI - Effects of the dopamine reuptake inhibitor PTT on reinstatement and on food- and cocaine-maintained responding in rhesus monkeys. AB - RATIONALE: High-affinity, slow-onset, long-acting dopamine transporter (DAT) inhibitors are being considered as potential agonist replacement therapies for cocaine addiction, and therefore the ability of these drugs to reinstate cocaine seeking and to selectively decrease cocaine-maintained responding should be assessed. OBJECTIVES: The purpose of these experiments was to evaluate the effects of the active enantiomer of a high-affinity, slow-onset, long-acting DAT inhibitor, (-)2beta-propanoyl-3beta-(4-tolyl)-tropane (PTT), and cocaine on food- and cocaine-maintained responding and on extinguished responding previously maintained by cocaine in non-human primates using a within-subjects design. METHODS: Rhesus monkeys (n=3) responded under a multiple fixed-ratio schedule of food (1 g) and drug reinforcement, and cocaine dose-response curves (saline, 0.003-0.3 mg/kg per injection) were determined. The effects of pretreatment with (-)PTT (0.001-0.056 mg/kg, i.v.) and cocaine (0.03-0.3 mg/kg, i.v.) were determined when the dose of cocaine that maintained peak response rates (0.03 mg/kg per injection) or saline was available. RESULTS: (-)PTT and cocaine reduced cocaine intake; (-)PTT affected cocaine self-administration only at doses that also decreased food-maintained responding. (-)PTT and cocaine reinstated responding that was previously reinforced by cocaine at lower doses than were necessary to decrease cocaine-maintained responding. For all studies, PTT was at least 1.0 log-unit more potent than cocaine. Compared to cocaine, PTT had a longer duration of action in all behavioral measures. CONCLUSIONS: These results suggest that PTT would decrease cocaine use, but only at doses that disrupted other behaviors. It appears that the potency of this class of drugs to reinstate cocaine-seeking is substantially greater than their potency at decreasing cocaine self-administration. PMID- 14735292 TI - Brain concentrations of d-MDMA are increased after stress. AB - RATIONALE: In the mouse but not the rat, d-3,4-methylenedioxymethamphetamine (d MDMA) is a dopaminergic neurotoxicant. Various stressors and hypothermia protect against d-MDMA-induced neurotoxicity through unknown mechanisms, one of which could be a reduction in the distribution of d-MDMA to the brain. OBJECTIVES: We determined striatal levels of d-MDMA in relation to body temperature in mice exposed to a neurotoxic regimen of d-MDMA in the presence or absence of various stressors. METHODS: Female C57BL6/J mice received a neurotoxic regimen of d-MDMA (15.0 mg/kg s.c. as the base every 2 hx4) alone or in combination with manipulations with a known neuroprotective status. d-MDMA levels were determined by HPLC with fluorometric detection while rectal temperature provided core temperature status. Levels of dopamine, tyrosine hydroxylase and GFAP were used to assess neurotoxicity. RESULTS: Restraint, ethanol co-treatment and cold stress were neuroprotective, caused hypothermia and increased striatal d-MDMA levels by 4- to 7-fold. Corticosterone treatment, as a stress mimic, did not alter striatal d-MDMA or temperature and was not protective. The protective glutamate receptor antagonist, MK-801, doubled striatal d-MDMA levels and caused hypothermia. CONCLUSIONS: Although stress and other protective manipulations can alter the striatal concentration of d-MDMA their hypothermia-inducing properties appear a more likely determinant of their neuroprotection against the striatal dopaminergic neurotoxicity of d-MDMA. PMID- 14735293 TI - Effect of 5-HT depletion by MDMA on hyperthermia and Arc mRNA induction in rat brain. AB - RATIONALE: 3,4-Methylenedioxymethamphetamine (MDMA) administration to rats produces an acute hyperthermic response and induces localised neuronal activation, which can be visualised via expression of immediate-early genes. The pharmacological and anatomical basis of these effects are unclear. At high doses, MDMA also causes selective neurotoxicity at serotonergic nerve terminals. OBJECTIVE: We investigated the effect of 5-hydroxytryptamine (5-HT) depletion on the acute hyperthermic response to MDMA and the pattern of neuronal excitation indicated by Arc (activity-regulated cytoskeleton associated gene) in naive rats and following administration of MDMA at a neurotoxic dose. METHODS: Expression of Arc mRNA was investigated by in situ hybridisation histochemistry using 35S labelled oligonucleotide probe. RESULTS: MDMA induced a significant hyperthermia together with increased Arc mRNA expression in cortical regions, caudate-putamen and CA1 hippocampus but not hypothalamus. At 21 days after a neurotoxic dose of MDMA, brain 5-HT and 5-HIAA levels were significantly reduced by 21-32%. In these animals, both the hyperthermic response and the pattern and extent of Arc mRNA expression induced by a subsequent dose of MDMA were unaltered. However, basal Arc expression was significantly increased in cortical regions and CA1 hippocampus. CONCLUSION: We conclude that the acute hyperthermic response induced by MDMA is not attenuated by moderate depletion of 5-HT, further questioning mediation via a serotonergic mechanism. Arc mRNA induction by MDMA exhibits highly localised expression, which is not altered following 5-HT depletion. However, following a neurotoxic dose of MDMA, basal expression of Arc is increased, particularly in cortex and CA1, suggesting that mechanisms underlying synaptic plasticity might also be modified. PMID- 14735294 TI - Visuospatial memory impairments in users of MDMA ('ecstasy'). AB - RATIONALE: Previous studies have presented conflicting findings regarding visuospatial span deficits in MDMA ('ecstasy') users, possibly attributable to a lack of distinction between simple visuospatial span and visuospatial working memory span. Both draw upon central executive processing, while the latter also involves concurrent goal-orientated visuospatial processing. OBJECTIVES: This study compared visuospatial working memory span for MDMA users and controls. An additional concurrent task also loading on the central executive tested for inter group differences related to central executive workload. METHOD: MDMA user group (25 current users, 10 previous users and 18 non-users) was between-participants, and dual task condition (concurrent alphabetic generation, random letter generation, and no dual task) was within-participants. The visuospatial working memory task required participants to serially recall a spatial sequence while simultaneously completing a visual judgement task, and was completed on its own and under dual task conditions. RESULTS: Overall, non-users performed significantly better than both MDMA user groups. However, contrary to expectation, the performance decrement among users was no worse with concurrent random generation than under control conditions. Analyses controlling for background variables and the use of other drugs in the previous 3 months showed that the main effect of MDMA remained significant following control for intelligence, alcohol, amphetamines and cocaine, among other potential confounds. Unclear results were found following control for cannabis use. CONCLUSIONS: The MDMA users experienced deficits in visuospatial working memory span. The lack of interaction between dual task condition and user group may be due to inter-group differences in central executive utilisation under different task conditions. PMID- 14735296 TI - Reduction of lipopolysaccharide-induced cyclooxygenase-2 expression in diabetic arteries. AB - In order to clarify the effects of diabetes mellitus on induction of cyclooxygenase (COX)-2 and subsequent prostaglandin production in blood vessels, we investigated lipopolysaccharide (LPS)-induced COX-2 induction and prostacyclin production in aortic strips isolated from streptozotocin-induced diabetic rats and vehicle-injected control rats. The rats at 10 weeks after injection of streptozotocin had significantly lower body weights and higher serum glucose levels compared with those in the control rats. LPS stimulation resulted in a marked increase in the release of prostacyclin from the aortic strips. This increase was abolished in the presence of indomethacin or cycloheximide but was not affected by removal of the endothelium. In diabetic aortae, both LPS-induced prostacyclin production and COX-2 induction were diminished compared with the control aortae. No significant difference in COX-1 expression was observed between diabetic and control aortae. The diminution of LPS-induced COX-2 expression was also observed in alveolar macrophages isolated from diabetic rats. These results suggest that COX-2 expression and subsequent prostacyclin production in response to LPS are selectively attenuated in diabetic blood vessels. PMID- 14735295 TI - Quetiapine reduces nocturnal urinary cortisol excretion in healthy subjects. AB - RATIONALE: Hypothalamic-pituitary-adrenal (HPA) axis dysfunction is a frequent finding in psychiatric disorders, including psychotic depression and schizophrenia. Conflicting results exist concerning the influence of antipsychotics on the HPA-axis. OBJECTIVE: Therefore, this double-blind, placebo controlled, randomized cross-over study investigated the effect of quetiapine on nocturnal urinary cortisol and melatonin excretion in 13 healthy male subjects under conditions of undisturbed and experimentally disturbed sleep. METHODS: Volunteers were studied 3 times for 3 consecutive nights (N0, adaptation; N1, standard sleep conditions; N2, acoustic stress) 4 days apart. Placebo, quetiapine 25 mg or quetiapine 100 mg was administered orally 1 h before bedtime on nights 1 and 2. Urine produced during the 8-h bedtime period was collected for later determination of cortisol and melatonin concentrations by standard radioimmunoassays. RESULTS: MANOVA showed a significant effect for N1 vs. N2 with elevated total amount of cortisol ( p<0.005) and melatonin ( p<0.05) excretion after acoustic stress. Both quetiapine 25 mg and 100 mg significantly ( p<0.0005) reduced the total amount of cortisol excretion in comparison to placebo. No interaction effect of stress condition was observed. There was no effect of quetiapine on melatonin levels. CONCLUSION: The significant reduction of nocturnal cortisol excretion following quetiapine reflects a decreased activity of the HPA-axis in healthy subjects. This finding may be an important aspect in quetiapine's mode of action in different patient populations. PMID- 14735297 TI - Genes involved in the anaerobic degradation of toluene in a denitrifying bacterium, strain EbN1. AB - The organization of all genes required for the anaerobic conversion of toluene to benzoyl-CoA was investigated in denitrifying Azoarcus-like strain EbN1. All of these genes are clustered within 25.3 kb of contiguous DNA sequence, which includes only a few intervening sequences. The toluene-catabolic genes are organized in two apparent operons. One contains the genes ( bssCAB) for the three subunits of benzylsuccinate synthase, which initiates anaerobic toluene degradation by converting toluene to ( R)-benzylsuccinate. The BssCAB proteins of strain EbN1 are most similar to those of Thauera aromatica strain K172. The bssCAB genes are part of a larger putative operon ( bssDCABEFGH), which contains the gene bssD, encoding the activase for benzylsuccinate synthase, and four genes ( bssEFGH) encoding proteins of unknown function. RT-PCR experiments showing continuation of transcription over the three largest intergenic regions of the bss operon support the assumed structure. Moreover, BssG was identified as toluene-induced protein. Downstream of the bss genes, another large putative operon ( bbsA- H) was identified that contains all genes required for beta oxidation of benzylsuccinate to benzoyl-CoA, e.g. bbsEF, encoding succinyl-CoA:( R)-benzylsuccinate CoA-transferase. Immediately upstream of the bss operon, genes for a two-component regulatory system were identified; their products may sense toluene and induce the expression of both catabolic operons. The order and sequences of the bss and bbs genes are highly similar among toluene-degrading denitrifiers. The bss and bbs genes of the Fe(III)-reducing Geobacter metallireducens display less sequence similarity and are organized differently. The genes between the bss and bbs operons and in the flanking regions differ between strain EbN1 and the other strains. PMID- 14735298 TI - In Saccharomyces cerevisiae, the effect of H2O2 on ATP, but not on glyceraldehyde 3-phosphate dehydrogenase, depends on the glucose concentration. AB - As has been previously shown, Saccharomyces cerevisiae grown in 2% or 0.025% glucose uses this carbohydrate by the fermentative or oxidative pathways, respectively. Depending on the glucose concentration in the medium, the effect of the addition of H2O2 on the level of ATP and on glyceraldehyde-3-phosphate dehydrogenase (GAPDH) activity differed. In the presence of 2% glucose, ATP and GAPDH decreased sharply during the first few minutes of treatment, whereas in the presence of 0.025% glucose, GAPDH activity decreased similarly, but the ATP level remained practically unchanged. The addition of 3 mM glutathione to the culture media prevented the depletion of ATP levels and GAPDH activity in the presence of H2O2. Catalase and superoxide dismutase activities did not vary significantly when yeast cells were grown either in 2% or in 0.025% glucose. PMID- 14735299 TI - Risk-taking, coordination and upper limb fractures in children: a population based case-control study. AB - The aim of this population based case-control study was to examine the association between risk-taking behaviour, motor coordination and upper limb fractures in children aged 9-16 years. A total of 321 fracture cases and 321 randomly selected individually matched controls were studied. The number for different types of upper limb fractures was 91 for hand, 190 for wrist and forearm and 40 for upper arm. Risk-taking behaviour was determined by a 5-item interview-administered questionnaire. Motor coordination was assessed by the 8 point movement ABC that tests manual dexterity, ball skills as well as static and dynamic balance. Bone mass was assessed by dual energy X-ray absorptiometry (DXA) and metacarpal morphometry. In general, there was heterogeneity by fracture site with regard to associations. Risk-taking behaviour was associated with hand fracture risk but not other fracture sites for downhill cycling behaviour (OR: 2.0/category, 95% CI: 1.1-3.7), dare behaviour (OR: 3.3/category, 95% CI: 1.1 10.0) and total risk-taking score (OR: 2.6/category, 95% CI: 1.3-5.7). Conversely, coordination measures were associated with wrist and forearm fractures only: cutting/threading (OR: 1.2/unit, 95% CI: 1.0-1.4); flower trail (OR: 1.2/unit, 95% CI: 1.0-1.4) and dynamic balance score (OR: 1.1/unit, 95% CI: 1.0-1.2). Backward stepwise analysis selected total risk taking score for hand fracture, and dynamic balance score for wrist and forearm fracture. None of the risk-taking or coordination scores were associated with upper arm fractures. These associations were unchanged following adjustment for bone mass. In conclusion, the propensity to take risks is most strongly associated with hand fracture risk while dynamic balance is most strongly associated with wrist and forearm fracture risk in children. These results inform the development of fracture prevention strategies in children. PMID- 14735300 TI - Should age influence the choice of quantitative bone assessment technique in elderly women? The EPIDOS study. AB - In a prospective cohort of 7,598 women aged 75 and over, we analyzed the effect of age on the ability of femoral neck bone mineral density (BMD) and of ultrasound (BUA and SOS) of the calcaneus to predict hip fracture. Unadjusted regression analysis showed that the risk of hip fracture was increased 1.7 times for one standard deviation increase in age (3.7 years). Overall, for a decrease of one standard deviation in quantitative bone measures, the risk was significantly increased by 2.2 times for BMD (1.9-2.5), 1.8 for BUA (1.6-2.1), and 1.9 for SOS (1.6-2.2). However the average relative risk associated with a decrease in BMD tends to diminish with advancing age, meaning that a smaller part of the risk is explained by BMD in the very elderly. This is confirmed by the areas under the ROC curves (AUC) of BMD that are significantly better before 80 years (0.75 [0.73-0.76]) than after (0.65 [0.63-0.67] in group 80-84 years and 0.65 [0.61-0.68] in group >/=85). On the other hand, as the absolute risk increases exponentially with age, the number of hip fractures attributable to a low BMD is still important in the very elderly, the risk difference between the lowest and the highest quartile of BMD is 25 hip fractures / 1,000 woman-years in the group >/=85 compared with 16 in the two other groups. Thus, after 80, quantitative assessment of bone may still be of interest for clinical decisions. Compared with quantitative ultrasound parameters, the ability of BMD to predict hip fracture was significantly superior to that of BUA and SOS only before the age of 80 (AUC of BMD 0.75 [0.73-0.76], BUA 0.67 [0.66-0.69], SOS 0.67 [0.65 0.69]). For patients older than 80, we did not observe significant differences in AUC between DXA and QUS to predict hip fracture. PMID- 14735301 TI - Knowledge about osteoporosis in a cohort of Polish females: the influence of age, level of education and personal experiences. AB - In this study, a group of 1065 women aged 16-72 years recruited from patients attending general practitioners was studied to investigate knowledge of osteoporosis and attitude towards methods of preventing the disease. The interviews were carried out by students of the Silesian School of Medicine, using a structured questionnaire. The average number of correct answers in the whole population was 7.05, and in women with established osteoporosis 6.89. In the whole population, the majority of answers were correct and ranged from 60% to 95% in seven out of ten questions. The answers for three questions were incorrect in about half or more of the cohort: 53% of subjects considered that osteoporosis could be cured; for 50% of women osteoporosis is a minor health problem (except for the youngest women and women with university education--74% and 69% of correct answers, respectively); and 58% of women considered that those with osteoporosis should not engage in physical activity due to the risk of falling and causing a fracture. A simple chi-squared test was used to show the role of age, level of education and personal experiences with osteoporosis on answers given by the subjects studied. Age (six out of ten answers), level of education (seven out of ten answers) and personal experience (four out of ten answers) significantly affected answers given. The number of correct answers decreased with age and increased with level of education, and there was no systematic influence of personal experience. In summary, data collected provide important information about knowledge of osteoporosis. Generally, the level of knowledge about osteoporosis was high. Higher level of education and younger age improve the knowledge of osteoporosis with no systematic influence of personal experience with the disease. PMID- 14735302 TI - Simplified MRI sequences for postoperative control of hamstring anterior cruciate ligament reconstruction. AB - INTRODUCTION: Usually, standard radiographs are used for postoperative quality follow-up after ACL reconstruction. However, with the use of hamstring grafts and bioabsorbable implants, accurate assessment of the tunnel and implant position is impossible. The graft and its relation to anatomical landmarks cannot be evaluated directly. MRI is an alternative to radiography, permitting direct graft visualization and 3-dimensional assessment of the tunnel position, but it is expensive and time consuming for routine use. The aim of this study was to develop a simplified MRI protocol and to evaluate it for routine postoperative quality follow-up after ACL reconstruction. MATERIALS AND METHODS: Various scanning protocols were tested in a series of 105 patients and evaluated for image sharpness, clarity of the structures, susceptibility to artefacts, applicability regarding precise analysis of graft and tunnel position, and time consumption. One simplified specific scan protocol was then defined and applied in a series of 60 consecutive patients after hamstring ACL replacement. The position of the femoral and tibial tunnels was measured in the sagittal, coronal and axial sections and classified according to Harner (femoral) and Staubli (tibial). Impingement of the graft in the intercondylar roof was analysed according to Howell. The position of the bioabsorbable interference screws was assessed. RESULTS: Scan protocol: T2-weighted gradient-echo sequences (GRE) with TR 246 ms, TE 11 ms, flip angle 25 degrees, 2 mm sections and a 256 x 256 matrix yielded the best image quality of tendon grafts and bone tunnels with tolerable time consumption (average scanning time per patient 1 min 40 s). Altogether 8-16 sections were obtained for every patient. Tunnel placement: 46/60 (77%) of the femoral tunnels were in zone 4, 13/60 (21%) at the border of zones 3 to 4, 1/60 (2%) in zone 3 in the sagittal plane (Harner). The femoral tunnels in the axial plane were at 10:30 o'clock in 32/60 (53%), at 11:00 o'clock at 24/60 (40%) and at 10:00 o'clock in 4/60 (4%) patients. The mean distance of the anterior border of the tibial tunnel from the anterior cortex was 39% (+/- 4.9%) related to the total sagittal diameter of the tibia. There was no graft impingement. The position of the interference screws was anterior to the grafts in all cases. CONCLUSION: Simplified MRI sequences can be used for postoperative quality follow up after ACL replacement and are an alternative to standard radiographs giving more specific and precise information regarding tunnel position and screw placement. Analyzing the bone tunnels in a series of 60 patients demonstrated that correct assessment of tunnel placement after arthroscopic ACL reconstruction is feasible using this simplified MRI technique. PMID- 14735303 TI - Bilateral and simultaneous rupture of the triceps tendons in chronic renal failure and secondary hyperparathyroidism. AB - BACKGROUND: We report a case of a 27-year-old (male) patient with bilateral and simultaneous rupture of the triceps tendons. He was suffering from chronic renal failure secondary to acute glomerulonephritis and for 4 years he underwent maintenance hemodialysis three times a week. This injury is extremely rare, and only one case has been reported in the international literature. Based on previous cases of tendon ruptures in patients with chronic renal failure, we believe that secondary hyperparathyroidism is the primary causative factor in his case. CASE REPORT: A primary repair was performed using heavy, nonabsorbable sutures passed through holes drilled in the olecranon with the technique described by Levy. The suture line was then protected with a figure-of-eight tension band wire. Our patient obtained full range of motion bilaterally within 3 months. PMID- 14735304 TI - Nucleolar characteristics of reducing bodies in reducing body myopathy. AB - Reducing body myopathy is a rare muscle disease defined by abnormal inclusions in affected muscle fibers that can be stained with menadione-nitroblue tetrazolium. The origin of these inclusions has not been determined. Here we show that reducing bodies bear characteristics of nucleoli. Ultrastructurally, muscle biopsy specimens of a patient with adult-onset reducing body myopathy showed granular structures of reducing bodies with features similar to the granules of nucleoli, which consisted of pre-ribosomes. In addition, reducing bodies were positive for histochemistry of argyrophilic nucleolar organizer regions (a method for detecting the areas where ribosomal RNA is actively transcribed in the nucleolus), and for antibodies against nucleoli and nuclear ribonucleoprotein. The current findings suggest that reducing bodies contain pre-ribosomes and their associated proteins of the nucleolus and that formation of reducing bodies may result from defects of processing and assembly of ribosomes. PMID- 14735305 TI - Redistribution of aquaporin-4 in human glioblastoma correlates with loss of agrin immunoreactivity from brain capillary basal laminae. AB - Vasogenic edema is one of the most serious clinical problems in brain tumors and tightly connected to water shifts between the different fluid compartments in the brain. Aquaporin water channels have been recognized to have an important impact on the development of edematous swelling in the brain. Astrocytes, which are believed to induce or at least maintain the blood-brain barrier in the brain capillary endothelial cells, express the aquaporin isoform AQP4. Normally, AQP4 is highly concentrated in the glial membrane where astrocytes contact mesenchymal space, such as perivascular or brain superficial regions. Parenchymal membranes do not show any immunocytochemical AQP4-specific signal. We investigated the AQP4 expression in human glioblastoma and correlated it with the expression pattern of the extracellular heparan sulfate proteoglycan agrin and members of the dystrophin-dystroglycan complex. We found that AQP4 completely covered the surface of the glioma cells. alpha-Dystroglycan was absent from glial membranes but retained in endothelial membranes. Utrophin and dystrophin remained restricted to the endfoot membrane in those cells in which AQP4 had been redistributed, whereas alpha-syntrophin redistributed together with AQP4 across the entire cell surface. Since alpha-dystroglycan operates as a binding protein for agrin, these observations support the suggestions that (1) AQP4 is tightly associated with the dystrophin-dystroglycan complex, and (2) agrin is necessary for the polarized distribution of AQP4 in the astrocyte. The results are discussed in connection with the fact that normally AQP4 is assembled in the so called orthogonal arrays of particles (OAPs). The restriction of AQP4/OAPs to the endfoot membrane may be dependent on the presence of agrin, and this might be essentially connected to the ability of astrocytes to maintain the integrity of the blood-brain barrier. PMID- 14735306 TI - Intrathoracic and intraabdominal locations of a cystic benign tumor: congenital etiology due to embryological diaphragm development? AB - A rare case of a benign cystic two-cavity tumor with intrathoracic and intraabdominal localisation is presented. The tumor's embryological etiology, embryological development of the diaphragm and the occurrence of embryonic tumors in general, are discussed. To our knowledge this is the first documented case of a benign two-cavity tumor in childhood and infancy. PMID- 14735308 TI - Mechanisms of stick insect locomotion in a gap-crossing paradigm. AB - Locomotion of stick insects climbing over gaps of more than twice their step length has proved to be a useful paradigm to investigate how locomotor behaviour is adapted to external conditions. In this study, swing amplitudes and extreme positions of single steps from gap-crossing sequences have been analysed and compared to corresponding parameters of undisturbed walking. We show that adaptations of the basic mechanisms concern movements of single legs as well as the coordination between the legs. Slowing down of stance velocity, searching movements of legs in protraction and the generation of short steps are crucial prerequisites in the gap-crossing task. The rules of leg coordination described for stick insect walking seem to be modified, and load on the supporting legs is assumed to have a major effect on coordination especially in slow walking. Stepping into the gap with a front leg and antennal contact with the far edge of the gap provide information, as both events influence the following leg movements, whereas antennal "non-contact" seems not to contain information. Integration of these results into the model of the walking controller can improve our understanding of insect locomotion in highly irregular environments. PMID- 14735307 TI - Comparative aspects of natriuretic peptide physiology in non-mammalian vertebrates: a review. AB - The natriuretic peptide system is a complex family of peptides and receptors that is primarily linked to the maintenance of osmotic and cardiovascular homeostasis. A natriuretic peptide system is present in each vertebrate class but there are varying degrees of complexity in the system. In agnathans and chondrichthyians, only one natriuretic peptide has been identified, while new data has revealed that multiple types of natriuretic peptides are present in bony fish. However, it seems in tetrapods that there has been a reduction in the number of natriuretic peptide genes, such that only three natriuretic peptides are present in mammals. The peptides act via a family of guanylyl cyclase receptors to generate the second messenger cGMP, which mediates a range of physiological effects at key targets such as the gills, kidney and the cardiovascular system. This review summarises the current knowledge of the natriuretic peptide system in non mammalian vertebrates and discusses the physiological actions of the peptides. PMID- 14735309 TI - A stingless bee uses labial gland secretions for scent trail communication ( Trigona recursa Smith 1863). AB - The pheromones used by several species of stingless bees for scent trail communication are generally assumed to be produced by the mandibular glands. Here we present strong evidence that in Trigona recursa these pheromones originate from the labial glands, which are well developed in the heads of foragers. Analysis of the behavior involved in scent marking shows that a bee extends her proboscis and rubs it over the substrate. A single scent marking event lasts for 0.59+/-0.21 s while the bee runs a stretch of 1.04+/-0.37 cm on a leaf. According to choice experiments the bees are attracted by a feeder baited with labial gland extract (84.2+/-6% of the bees choose this feeder) but repelled from a feeder baited with mandibular gland extract (only 27.5+/-13.1% of the bees choose this feeder). They do not discriminate between two clean feeders (49.6+/-3% of the bees at a feeder). 87+/-5.1% of bees already feeding leave the feeder after the application of mandibular gland extract whereas only 6.2+/-4.9% and 2.6+/-4% do so when labial gland extract or pure solvent was applied. PMID- 14735310 TI - The role of [(18)F] FDG-PET, CT/MRI and tumor marker kinetics in the evaluation of post chemotherapy residual masses in metastatic germ cell tumors--prospects for management. AB - The purpose of this study was to assess the ability of [(18)F]FDG-PET, CT/MRI and serum tumor marker (TM) to predict the viability of residual masses after high dose chemotherapy (HD-Ctx) in patients with metastatic germ cell tumors (GCT). In a prospective study, 60 residual tumors in 28 GCT patients were classified as viable/nonviable by FDG-PET, CT/MRI and TM levels. The results were validated either by histological examination of a resected mass and/or biopsy or by clinical/radiological follow-up for at least 6 months. There were no significant differences among the sensitivities observed with PET, CT/MRI and TM, but PET was significantly more specific than CT/MRI in predicting residual mass viability. TM showed the highest specificity. The highest accuracy in classification of residual tumors was achieved by a combination of PET, CT/MRI and TM (area under the ROC curve =0.91). All mature teratomas showed false-negative PET results with SUVs in the same range as necrosis. For classification of residual masses after HD-Ctx of metastatic GCT, [(18)F]FDG-PET is a valuable diagnostic method to complement the established procedures CT and TM. Positive PET results are highly correlated with the presence of viable tumor, but residual masses with negative PET findings still require resection. In cases of tumor progression diagnosed by CT and elevated TM, additional PET examinations are without benefit. PET seems useful in patients with stable disease or partial remission in CT/MRI and normalized TM as well as in marker-negative disease. PMID- 14735312 TI - Effect of light on the organogenic ability of garlic roots using a one-step in vitro system. AB - A simple and efficient garlic in vitro shoot regeneration protocol has been developed. This system uses axenic root tips cultivated from the beginning in the presence of light and does not require any change or refreshing of the original medium during the entire process. The application of light from the beginning of the culture process did not affect the callus formation rate but did significantly improve the explant regeneration ability. In a 2-month period it was possible to obtain up to 250 shoots per gram of callus. PMID- 14735311 TI - Dicistronic expression of the green fluorescent protein and antibiotic resistance genes in the plastid for selection and tracking of plastid-transformed cells in tobacco. AB - A plastid transformation vector was constructed for dicistronic expression of the aminoglycoside 3'-adenyltransferase (aadA) and green fluorescent protein (gfp) genes under the control of the plastid rrn promoter. Gold particles coated with the vector DNA were bombarded onto tobacco leaf explants using a particle delivery system. Leaf explants produced adventitious shoots when cultured on shoot-inducing medium containing 500 mg l(-1) spectinomycin. Shoots that exhibited green fluorescence under UV light were selected. Southern blot analysis detected the presence of the aadA and gfp genes between trnA and trnI in the plastid genome. Northern blot analysis revealed that the aadA and gfp genes were both properly transcribed into a dicistronic transcriptional unit. The expression of the gfp gene in the plastid enabled separation of transformed chloroplasts from wild-type chloroplasts in the protoplast under a fluorescent microscope. The overall results indicate that dicistronic expression of the aadA and gfp genes in the plastid simplifies gene manipulation, facilitating selection and tracking of plastid-transformed cells. PMID- 14735313 TI - Polyethylene glycol and abscisic acid improve maturation and regeneration of Panax ginseng somatic embryos. AB - Embryogenic culture was initiated from mature zygotic embryos of Panax ginseng. Multiple somatic embryos formed and proliferated on Murashige and Skoog medium supplemented with 2,4-dichlorophenoxyacetic acid (2.26 microM) and kinetin (0.046 microM). Mature as well as immature somatic embryos grew into plantlets lacking roots on the same media. Histomorphological analysis of somatic embryos treated with abscisic acid (ABA) and polyethylene glycol (PEG 4000) showed a slight improvement in the root meristem organization of torpedo-stage embryos (embryos were more compact and their cells exhibited a lower degree of vacuolation). Shoot regeneration of non-treated somatic embryos was 31% while that for somatic embryos treated with PEG 4000 and ABA was 70%. Moreover, 75% of plants regenerated from PEG- and ABA-treated embryos formed roots while plants from non treated embryos did not form roots. PMID- 14735314 TI - Characterisation of the Mucor circinelloides regulated promoter gpd1P. AB - The promoter of the Mucor circinelloides gpd1 gene encoding glyceraldehyde-3 phosphate dehydrogenase (gpd1P) was recently cloned and used for the production of recombinant proteins, such as the Aspergillus niger glucose oxidase 1 (GOX). This represents the first example of the application of a strong and regulated promoter from this fungus for recombinant protein production. The original 741-bp gpd1P promoter fragment conferred hexose-dependent expression of GOX in M. circinelloides. To understand the regulatory mechanisms involved in gpd1P-driven expression and to develop improved promoter fragments, deletion derivatives of gpd1P were constructed. These derivatives were fused to the A. niger gox1 gene and used to construct strains containing a single copy of the expression cassette. GOX activity was detected in strains containing the full-length gpd1P and also in strains containing a 713-bp or a 361-bp derivative. Expression levels for the 361-bp derivative were high and comparable, regardless of the carbon source used. This promoter represents a useful derivative for constitutive heterologous gene expression in M. circinelloides. PMID- 14735315 TI - Renal artery embolization: a first line treatment option for end-stage hydronephrosis. AB - Conventionally poorly functioning hydronephrotic kidneys have been removed if they are symptomatic. In our unit, patients are offered renal artery embolization as an alternative treatment option. PATIENTS AND METHODS: Fifteen patients (11 male, 4 female) with a mean age of 32.9 yr (20-51 yrs) have undergone renal artery embolization for symptomatic hydronephrosis with poor function. Mean follow-up was 64.13 weeks (range 14-200). All patients had loin pain and hydronephrosis. Twelve patients had primary pelvi-ureteric junction obstruction (PUJO). Two patients had poorly functioning hydronephrotic kidneys secondary to chronic calculous obstruction. One patient had chronic pain in an obstructed but reasonably functioning kidney following a previous pyeloplasty for PUJO which demanded intervention. Mean split function on renography was 11% (range 0-46%). Selective renal artery embolization was carried out under antibiotic cover using a 7 Fr balloon occlusion catheter and absolute alcohol, steel coils, and polyvinyl alcohol particles. RESULTS: Nine patients developed post-embolization syndrome of self-limiting pain and pyrexia with no evidence of sepsis. One patient required readmission with this condition. One patient developed a hematoma at the puncture site. Mean hospital stay was 2.3 days. Fourteen patients are happy with the result and are completely pain free. One patient has minor discomfort but is delighted with the result. Nine patients have had follow-up ultrasound confirming resolution of the hydronephrosis. CONCLUSION: Renal artery embolization is an effective, safe, well-tolerated minimally invasive treatment option in end-stage hydronephrosis and we routinely offer it as an alternative to nephrectomy. PMID- 14735318 TI - Identifying biomarkers and surrogates of tumors (cancer biometrics): correlation with immunotherapies and immune cells. AB - The presence of inflammatory cells within cancer has been described for quite some time by pathologists, with generally improved outcome associated with their presence in various epithelial neoplasms. Most remarkably, this has included dendritic cells and T cells but more recently NK cells as well. Coupled with the rapid evolution of molecular technology, microarray analyses of primary tumors, serum and tumor proteomics, tumor capture analyses in the peripheral blood (together with quantitative RT-PCR), and novel histochemical markers and tissue microarrays, this provides the opportunity to establish a more effective means to study and classify into subsets various forms of cancer. Much of the current controversy in cancer diagnosis and pathologic assessment of prognosis lies in the application of these techniques in concert with other molecular tools including DNA microarrays, expression of histochemically defined cytokines, proangiogenic factors, and oncogene products, and correlating this with clinical relevance. Molecular detection technologies such as reverse transcriptase polymerase chain reaction, proteomics, and microarray analyses will be validated based on their integration with conventional cancer pathology and cancer diagnostics. Further work is needed to establish which cancer biomarkers and surrogates should be routinely measured and in which settings, and determining the appropriate sample size for such assays that can be validated in retrospective and prospective clinical studies. The ability to integrate these rapidly evolving strategies will consume much of our coordinate effort in cancer and cancer therapeutics for the near future. PMID- 14735319 TI - Enhancement of cytotoxic T-lymphocyte responses in patients with gastrointestinal malignancies following vaccination with CEA peptide-pulsed dendritic cells. AB - Carcinoembryonic antigen (CEA) is strongly expressed in a vast majority of gastrointestinal carcinomas. Recently, epitope peptides of CEA were identified. We have demonstrated HLA-A24-restricted peptide, CEA652[9] (TYACFVSNL), was capable of eliciting specific cytotoxic T lymphocytes (CTLs) which could lyse tumor cells expressing HLA-A24 and CEA. HLA-A24 is the most applicable MHC class I allele in the Japanese population. In this pilot study, we have used the peptide-pulsed dendritic cells (DCs) generated from peripheral blood mononuclear cells (PBMCs) supplemented with GM-CSF and IL-4 as the source of the vaccine. Eight patients with advanced CEA-expressing gastrointestinal malignancies received subcutaneous injections every 2 or 3 weeks. Immunomonitoring was performed by ELISpot (enzyme-linked immunosorbent spot) assay to measure the precursor frequency of CTLs and their capacity to elicit antitumor CTLs in vitro. Four of seven patients have developed their CTL response after vaccinations. DTH reaction was observed in one of eight patients at the DC-injected site. Skin biopsy at the injected site showed the infiltration of the lymphocytes. Furthermore, A24/CEA peptide tetramer assay revealed an increase in peptide specific T-cell precursor frequency in vaccinated patients. No significant toxic adverse effects were observed, except for mild diarrhea in one case after three vaccinations. Three patients have shown stabilization of the disease after vaccinations. In conclusion, our results clearly demonstrated that our vaccination protocol was safe and might develop a CEA-specific CTL response in cancer patients. PMID- 14735320 TI - Acute renal failure as the first presenting sign of non-Hodgkin's lymphoma. PMID- 14735321 TI - Assessment of microbial populations in methyl ethyl ketone degrading biofilters by denaturing gradient gel electrophoresis. AB - Denaturing gradient gel electrophoresis (DGGE) analysis of polymerase chain reaction-amplified genes coding for 16S rRNA was used to assess differences in bacterial community structure as a function of spatial location along the height of two biofilters used to treat a model waste gas stream containing methyl ethyl ketone (MEK). One of the laboratory-scale biofilters was operated as a conventional continuous-flow biofilter (CFB) and the other was operated as a sequencing batch biofilter (SBB). Both biofilters, inoculated with an identical starting culture and operated over a period lasting more than 300 days, received the same influent MEK concentration and same mass of MEK on a daily basis. The systems differed, however, in terms of the fraction of time during which contaminated air was supplied and the overall operating strategy employed. DGGE analysis indicated that microbial community structures differed as a function of height in each of the biofilters. The DGGE banding patterns also differed between the two biofilters, suggesting that operating strategies imposed on the biofilters imparted a sufficiently large selective pressure to influence microbial community structures. This may explain, in part, the superior performance of the SBB over the CFB during model transient loading conditions, and it may open new possibilities for purposely manipulating the microbial populations in biofilters treating gas-phase contaminants in a manner that leads to more favorable treatment performance. PMID- 14735322 TI - Transformation of artemisinin by Cunninghamella elegans. AB - Semi-synthetic derivatives of the anti-malarial drug artemisinin hold great promise in the search for an effective and economical treatment of chloroquine resistant forms of malaria. Unfortunately, synthetic functionalization of the artemisinin skeleton is often tedious and/or impractical. We seek to utilize 7beta-hydroxyartemisinin, obtained from microbial transformation, as a semi synthetic precursor for the synthesis of novel 7beta-substituted artemisinin anti malarial agents. Here we employ liquid cultures of Cunninghamella elegans as a means for the rational and economical bioconversion of artemisinin to 7beta hydroxyartemisinin in 78.6% yield. In addition, there were three other bioconversion products: 7beta-hydroxy-9alpha-artemisinin (6.0%), 4alpha-hydroxy-1 deoxoartemisinin (5.4%), and 6beta-hydroxyartemisinin (6.5%). PMID- 14735324 TI - Reutericyclin: biological activity, mode of action, and potential applications. AB - Reutericyclin is an inhibitory compound produced by sourdough isolates of Lactobacillus reuteri that is structurally but not functionally related to naturally occurring tetramic acids. It is bacteriostatic or bactericidal to gram positive bacteria based on its activity as a proton-ionophore, and a broad range of food-related spoilage organisms and pathogens is inhibited by reutericyclin. Gram-negative bacteria are resistant to reutericyclin because of the barrier properties of their outer membrane, and resistance of beer-spoiling lactobacilli towards hop bitter acids provides cross-protection to reutericyclin. Remarkably, reutericyclin-producing strains were shown to persist for a period of 10 years in an industrial sourdough fermentation, and reutericyclin was shown to be produced in concentrations active against competitors during growth of L. reuteri in sourdough. Based on the known properties of reutericyclin and L. reuteri, reutericyclin-producing strains may have applications in the biopreservation of foods. Furthermore, these strains were shown to colonize reconstituted lactobacilli-free mice at high levels. Therefore, they could serve as a suitable model system to evaluate a possible impact of antimicrobial compounds on the intestinal microflora of humans and animals. PMID- 14735323 TI - Anaerobic degradation of monoaromatic hydrocarbons. AB - Over the last two decades significant advances have been made in our understanding of the anaerobic biodegradability of monoaromatic hydrocarbons. It is now known that compounds such as benzene, toluene, ethylbenzene, and all three xylene isomers can be biodegraded in the absence of oxygen by a broad diversity of organisms. These compounds have been shown to serve as carbon and energy sources for bacteria growing phototrophically, or respiratorily with nitrate, manganese, ferric iron, sulfate, or carbon dioxide as the sole electron acceptor. In addition, it has also been recently shown that complete degradation of monoaromatic hydrocarbons can also be coupled to the respiration of oxyanions of chlorine such as perchlorate or chlorate, or to the reduction of the quinone moieties of humic substances. Many pure cultures of hydrocarbon-degrading anaerobes now exist and some novel biochemical and genetic pathways have been identified. In general, a fumarate addition reaction is used as the initial activation step of the catabolic process of the corresponding monoaromatic hydrocarbon compounds. However, other reactions may alternatively be involved depending on the electron acceptor utilized or the compound being degraded. In the case of toluene, fumarate addition to the methyl group mediated by benzylsuccinate synthase appears to be the universal mechanism of activation and is now known to be utilized by anoxygenic phototrophs, nitrate-reducing, Fe(III) reducing, sulfate-reducing, and methanogenic cultures. Many of these biochemical pathways produce unique extracellular intermediates that can be utilized as biomarkers for the monitoring of hydrocarbon degradation in anaerobic natural environments. PMID- 14735326 TI - Identification of a novel polymorphism involving a CGG repeat in the PTCH gene and a genome-wide screening of CGG-containing genes. AB - Mutations in the human homologue of the Drosophila patched gene (PTCH) are responsible for the hereditary disorder called nevoid basal cell carcinoma syndrome (NBCCS). PTCH has a CGG triplet repeat located 4 bp upstream of the first methionine codon. Here we report a novel polymorphism involving the number of the CGG-repeat. The major allele (86.3%) contained a repeat size of seven, whereas the minor allele contained eight. No significant difference in the distributions of genotypes was observed between normal and NBCCS individuals. However, when the repeat was inserted between a heterologous promoter and the luciferase gene, the longer repeats tended to induce higher luciferase activities, suggesting that the repeat length potentially affects the levels of gene expression. A genome-wide screening revealed that 68 and 146 genes contained a CGG/CCG repeat in the coding region and in the 5'-untranslated region (5'-UTR), respectively. None of the genes had this repeat in 3'-UTR. Interestingly, the number of genes with a CGG repeat in the 5'-UTR was significantly higher than that with a CCG repeat in the 5'-UTR. The localization of a CGG/CCG repeat in PTCH is quite unique in that only four other genes have been found in which the repeat is localized up to 4 bp upstream of the first methionine. PMID- 14735325 TI - Association of beta2-microglobulin with the alpha3 domain of H-2Db heavy chain. AB - MHC class I molecules are heterotrimeric complexes composed of heavy chain, beta2 microglobulin (beta2m) and short peptide. This trimeric complex is generated in the endoplasmic reticulum (ER), where a peptide loading complex (PLC) facilitates transport from the cytosol and binding of the peptide to the preassembled ER resident heavy chain/beta2m dimers. Association of mouse MHC class I heavy chain with beta2m is characterized by allelic differences in the number and/or positions of amino acid interactions. It is unclear, however, whether all alleles follow common binding patterns with minimal contributions by allele-specific contacts, or whether essential contacts with beta2m are different for each allele. While searching for the PLC binding site in the alpha3 domain of the mouse MHC class I molecule H-2Db, we unexpectedly discovered a site critical for binding mouse, but not human, beta2m. Interestingly, amino acids in the corresponding region of another MHC class I heavy chain allele do not make contacts with the mouse beta2m. Thus, there are allelic differences in the modes of binding of beta2m to the heavy chain of MHC class I. PMID- 14735327 TI - Giant mature cyst formation following mesh repair of hernias: an underreported complication? AB - BACKGROUND: Numerous complications have been described following the implantation of synthetic meshes during hernia repair; one of the rarest, with only three reported cases, is giant mature fibrous cyst formation. Our clinical experience with this complication has led us to believe that it may be more common than previously thought. METHODS: Surgical operation notes and pathology archives were reviewed for the period January 1998-January 2002 to determine the prevalence of mature cyst formation following mesh repair of hernias. RESULTS: Out of 1100 hernia repair operations involving the use of synthetic meshes in our institution during the period of study, five developed histologically confirmed mature fibrous cysts giving a prevalence of 0.45% for this complication. CONCLUSION: The formation of a giant mature cyst following mesh repair of hernias is an underreported complication. PMID- 14735328 TI - Massive intrapericardial herniation of stomach following pericardial window. AB - Herniation of intra-abdominal contents into the pericardial cavity is rare. We describe one such case occurring after creation of a pericardioperitoneal window for drainage of a pericardial effusion. The diagnosis of an intrapericardial hernia should be considered in patients presenting with gastrointestinal and/or cardiorespiratory symptoms following surgical procedures involving the diaphragm. PMID- 14735329 TI - Traumatic abdominal wall hernia: a reappraisal. AB - Traumatic abdominal wall hernia, a rare cause of hernia, has a confusing clinical picture and requires a high index of suspicion for prompt diagnosis and management. Such hernias, if missed, can result in high morbidity and may prove fatal. Distinction from a pre-existing hernia is important as well. We report our experience in two such cases, which had presented in a span of 9 months, and submit a brief analysis of 50 reviewed cases. PMID- 14735331 TI - Early immunohistochemical and functional markers indicating radiation damage of the parotid gland. AB - In order to evaluate the correlation between functional impairment and changes in the expression pattern of immunohistochemical antibodies in the early phase of radiation-induced dysfunction of salivary glands, eight rabbits were scintigraphically examined prior to and 24 h after irradiation with 15 Gy. The parotid glands were studied using HE-staining, Ki-67, alpha-smooth muscle actin (ASMA) and tenascin-C antibodies at every scintigraphic examination. The results demonstrated a significant alteration in the 99mTc-pertechnetate uptake in all irradiated glands. HE-staining showed no relevant impairment of salivary gland tissue in this early phase. Immunohistochemically, we observed a marked re distribution of ASMA and tenascin-C as well as a reduction of the proliferating rate of acinar cells. This immunohistochemical change correlated with the functional impairment manifested scintigraphically. This study proves the possibility to assess disorders of salivary gland function with immunohistological antibodies as early as 24 h after irradiation and yields the prerequisites to prove the effects of radioprotective agents on salivary gland tissues. PMID- 14735332 TI - Modeling carbon monoxide dehydrogenase/acetyl-CoA synthase (CODH/ACS): a trinuclear nickel complex employing deprotonated amides and bridging thiolates. AB - Carbon monoxide dehydrogenase/acetyl-CoA synthase (CODH/ACS) utilizes a unique Ni M bimetallic site in the biosynthesis of acetyl-CoA, where a square-planar Ni ion is coordinated to two thiolates and two deprotonated amides in a Cys-Gly-Cys motif. The identity of M is currently a matter of debate, although both Cu and Ni have been proposed. In an effort to model ACS's unusual active site and to provide insight into the mechanism of acetyl-CoA formation and the role of each of the metals ions, we have prepared and structurally characterized a number of Ni(II)-peptide mimic complexes. The mononuclear complexes Ni(II) N, N'-bis(2 mercaptoethyl)oxamide (1), Ni(II) N, N'-ethylenebis(2-mercaptoacetamide) (2), and Ni(II) N, N'-ethylenebis(2-mercaptopropionamide) (3) model the Ni(Cys-Gly-Cys) site and can be used as synthons for additional multinuclear complexes. Reaction of 2 with MeI resulted in the alkylation of the sulfur atoms and the formation of Ni(II) N, N'-ethylenebis(2-methylmercaptoacetamide) (4), demonstrating the nucleophilicity of the terminal alkyl thiolates. Addition of Ni(OAc)(2).4H(2)O to3 resulted in the formation of a trinuclear species (5), while 2 crystallizes as an unusual paddlewheel complex (6) in the presence of nickel acetate. The difference in reactivity between the similar complexes 2 and 3 highlights the importance of ligand design when synthesizing models of ACS. Significantly,5 maintains the key features observed in the active site of ACS, namely a square planar Ni coordinated to two deprotonated amides and two thiolates, where the thiolates bridge to a second metal, suggesting that 5 is a reasonable structural model for this unique enzyme. PMID- 14735333 TI - NMR-validated structural model for oxidized Rhodopseudomonas palustris cytochrome c(556). AB - The structure of oxidized Rhodopseudomonas palustris cytochrome c(556) has been modeled after that of high-spin cytochrome c' from the same bacterium, the latter being the protein with the greatest sequence identity (35%) among all sequenced proteins in the genomes. The two proteins differ in the number of ligands to iron and in spin state, the former being six-coordinate low-spin and the latter five coordinate high-spin. In order to validate this modeled structure, several structural restraints were obtained by performing a restricted set of NMR experiments, without performing a complete assignment of the protein signals. The aim was to exploit the special restraints arising from the paramagnetism of the metal ion. A total of 43 residual-dipolar-coupling and 74 pseudocontact-shift restraints, which together sampled all regions of the protein, were used in conjunction with over 40 routinely obtained NOE distance restraints. A calculation procedure was undertaken combining the program MODELLER and the solution structure determination program PARAMAGNETIC DYANA, which includes paramagnetism-based restraints. The directions and magnitude of the magnetic susceptibility anisotropy tensor were also calculated. The approach readily provides useful results, especially for paramagnetic metalloproteins of moderate to large dimensions. PMID- 14735334 TI - Electron transfer in crystals of the binary and ternary complexes of methylamine dehydrogenase with amicyanin and cytochrome c551i as detected by EPR spectroscopy. AB - EPR studies of the methylamine dehydrogenase (MADH)-amicyanin and MADH-amicyanin cytochrome c551i crystalline complexes have been performed on randomly oriented microcrystals before and after exposure to the substrate, methylamine, as a function of pH. The results show that EPR signals from the redox centers present in the various proteins can be observed simultaneously. These results complement and extend earlier studies of the complexes under similar conditions that utilized single-crystal polarized absorption microspectrophotometry. The binary complex shows a blue copper axial signal, characteristic of oxidized amicyanin. After reaction of substrate with the MADH coenzyme tryptophan tryptophylquinone (TTQ), the binary complex exhibits an equilibrium mixture of oxidized copper/reduced TTQ and reduced copper/TTQ. radical, whose ratio is dependent on the pH. In the oxidized ternary complex, the same copper axial signal is observed superimposed on the low-spin ferric heme features characteristic of oxidized cytochrome c551i. After addition of substrate to the ternary complex, a decrease of the copper signal is observed, concomitant with the appearance of the radical signal derived from the semiquinone form of TTQ. The equilibrium distribution of electrons between TTQ and copper as a function of pH is similar to that observed for the binary complex. This result was essential to establish that the copper center retains its function within the crystalline ternary complex. At high pH, with time the low-spin heme EPR features disappear and the spectrum indicates that full reduction of the complex by substrate has occurred. PMID- 14735335 TI - A new dorsal modular fixation device allows a modified approach in cervical and cervico-thoracic neoplastic lesions. AB - INTRODUCTION: The aim of this study was to improve the management of cervical tumor osteolysis. A new modular rod-screw implant system for the posterior instrumentation of the occipito-cervical, cervical and cervico-thoracic spine (neon occipito cervical system, Ulrich, Germany) is available since 2000. K-wire guided pedicle screws are used, CT-guided instrumentation is possible. Previous studies have demonstrated increased biomechanical stability compared to established posterior cervical systems. METHODS: The cervical and cervico thoracic spine of 8 patients (6 males, 2 females, mean age 62 years, range 48-77 years) with osteolysis due to plasmocytoma (n=2), bronchial (n=2), mammary (n=2), esophageal (n=1) and pancreatic (n=1) carcinoma were instrumentated since June 2001. RESULTS: A stable fixation without loosening or failure of the fixation system was achieved in all cases. No impairment of the neurogical status was observed. CONCLUSION: Posterior instrumentation of the cervical spine including the occipito-cervical and the cervico-thoracic region with a new modular angle stable rod-screw implant system offers good stabilization and allows simultaneous decompression. Since tumor masses are predominantly located in the anterior portion of the spine blood loss can be reduced. PMID- 14735336 TI - Pruritus in a patient with advanced cancer successfully treated with continuous infusion of granisetron. AB - We present a case of a patient with advanced cancer affected by severe pruritus not related with cholestasis and/or opioid treatment successfully treated with subcutaneous continuous infusion of granisetron (3 mg/24 h diluted in normal saline via a disposable elastomeric infusion device). Confirmatory studies with a control group are warranted to confirm these preliminary results. PMID- 14735337 TI - Symptom masquerade: understanding the meaning of symptoms. AB - GOALS: Somatization refers to patients who transform distress and global suffering into pain and symptom expression. We have observed the opposite phenomenon in some outpatients seen for palliative care: patients who transform pain nociception into global suffering or other symptoms. The goal of this study is to describe the meaning of these patients' experiences that are not expressed as pain but seem to masquerade as other symptoms. PATIENTS AND METHODS: Hermeneutic phenomenological research methods guided this study with ten patients with advanced cancer. Interviews continued until data were saturated. RESULTS: Results included that all patients described frustration that they had outlived their usefulness. All expressed an awareness that they were dying, some very directly with sadness but appreciation for the support they received from family and friends, some with anger, and some who were actively fighting death awareness because they were not "ready" to die. Patients described themselves as strong and healthy and reluctant to take medications, both because this was a new and unacceptable behavior for them, and because they were averse to the side effects. CONCLUSION: In conclusion, careful evaluation of symptom experiences can ensure appropriate treatment. Patients with advanced cancer may not report pain even though pain medications often improve their other symptoms and well-being. Patients valued selection of medication that allowed them to remain alert and functional. The meaning of symptoms clearly altered patients' reports and limited treatments they found acceptable. Understanding these meanings is important to ensure that patients are treated in the most appropriate ways possible. PMID- 14735338 TI - Non-scheduled consultation in oncologic patients. How many of them are true emergencies? An observational prospective study. AB - Oncologic emergencies have been extensively described and clearly defined. In oncology daily practice, cancer patients seek non-scheduled medical care in situations they perceive as a medical emergency, but which may not be a true emergency. The aim of the study was to identify the main symptoms leading to a non-scheduled consultation (NSC) and their relationship to the type of cancer, and to evaluate whether the diagnosis at discharge of patients admitted as result of a NSC correlates with a true oncologic emergency. This was a prospective observational study. Between July 2002 and April 2003, 365 NSCs were recorded. The most frequent baseline diseases were breast cancer (70), lung cancer (67), gastrointestinal cancer (52), lymphoma (42) and ovarian cancer (22). The most common symptoms for consultation were: fever (84), pain (81), cutaneous manifestations (26), dyspnea (23), bleeding (16) and abdominal distention (16). Overall, 114 of 365 NSCs (31%) resulted in admission. The most frequent symptoms resulting in admission were fever (42), pain (16), dyspnea (11), vomiting (9), neurologic manifestations (7), abdominal distention (6) and anuria (6). At discharge, only 30 patients (26%) admitted after a NSC were diagnosed with a defined oncologic emergency: febrile neutropenia (13), intestinal occlusion (12), obstructive uropathy (4) and abdominal perforation (1). True emergencies were not the most frequent causes of NSC at our institution. PMID- 14735339 TI - Five-year outcome of laparoscopic and Lichtenstein hernioplasties. AB - BACKGROUND: Laparoscopic hernia repair has been proved superior to open repairs in terms of short-term results, but long-term results of laparoscopic and open mesh repairs have been lacking until recently. METHODS: A total of 123 patients were randomly allocated to two treatment groups comparing laparoscopic and Lichtenstein hernioplasties in three separate trials. The first and second trials compared small and large mesh used in transabdominal preperitoneal repairs, and the third study compared totally extraperitoneal hernioplasty with the Lichtenstein operation. A 5-year follow-up visit was scheduled to assess recurrencies, symptoms, and patient satisfaction. RESULTS: For the follow up evaluation, 121 (98.4%) of the patients were reached. There were five hernia recurrences in the laparoscopic group (small mesh) and two in the Lichtenstein group (difference, 5%; 95% confidence interval, -4-13%; p = 0.3). One patient who underwent the transabdominal preperitoneal polypropylene procedure underwent reoperation 3 years later because of dense small bowel adhesions at the inguinal surgical site. Chronic groin pain was more common after open operation (0 vs 4) patients (difference 7%; confidence interval, -0.4-16%; p = 0.04). Ten patients (16%) in the laparoscopic group and 12 (20%) in the open group reported discomfort or pain at the surgical site. CONCLUSIONS: Both laparoscopic and Lichtenstein hernioplasties have a low risk for hernia recurrence if proper mesh size is used. The patients who undergo hernioplasty with open mesh hernioplasty seem to experience chronic symptoms and pain more often than those managed with the laparoscopic procedure. PMID- 14735340 TI - Hematogenous tumor cell dissemination during colonoscopy for colorectal cancer. AB - BACKGROUND: It has long been suspected that mechanical influences may enhance the release of viable colorectal cancer cells into the circulation. The objective of this study was to determine the extent of hematogenous tumor cell spread in colorectal cancer patients during colonoscopy. METHODS: Peripheral venous blood samples were taken before and after colonoscopy from 44 patients with colorectal cancer. Blood samples were examined using a reverse-transcriptase polymerase chain reaction assay to amplify cytokeratin 20 transcripts. RESULTS: Eleven patients with colorectal cancer displayed circulating tumor cells before and after colonoscopy (25%), whereas tumor cells were detected in six of 44 patients (14%) only after the procedure (p = 0.03, McNemar's test: tumor cell detection before after colonoscopy). All control samples consistently tested negative. CONCLUSIONS: Mechanical forces may result in enhanced release of viable colorectal cancer cells into the circulation; however, the clinical significance of these results needs to be clarified. PMID- 14735341 TI - Hand-assisted laparoscopic surgery (HALS): a report of 150 procedures. AB - BACKGROUND: This study was performed to evaluate the (long-term) morbidity associated with hand-assisted laparoscopic surgery (HALS) for various indications. METHODS: HALS procedures for various indications were evaluated prospectively from 1995 to 2002. The primary outcome parameters were postsurgical complications and the development of incisional hernias. RESULTS: Twenty-six splenectomies, 51 hand-assisted laparoscopic donor nephrectomies (HLDN), 34 segmental bowel resections, 29 proctocolectomies, and 10 emergency colectomies were evaluated. A Kustner or Pfannenstiel incision was used for handport placement. Minor complications (i.e., wound complications, urinary tract infection) occurred in 15%, 12%, 26%, 7%, and 33% of the patients after, respectively, splenectomy, HLDN, bowel resection, proctocolectomy, and emergency colectomy. Major complications (i.e., hemorrhage, anastomotic leakage) occurred in 15% and 12% of the patients after, respectively, bowel resection and proctocolectomy. Incisional hernias occurred in six patients (4%), all after a wound complication in the Kustner incision. CONCLUSION: HALS is fast, safe, and feasible for various indications, especially HLDN and (procto-)colectomies. Little advantage can be expected when HALS is applied in splenectomy and segmental bowel (sigmoid) resection. PMID- 14735342 TI - Laparoscopic radiofrequency ablation of hepatic tumors: prospective clinical evaluation of ablation size comparing two treatment algorithms. AB - BACKGROUND: Radiofrequency ablation (RFA) is gaining increased acceptance for the local control of liver tumors. Essential for achieving local tumor control are reproducible volumes of ablation that encompass the tumor and a margin of normal liver parenchyma. The technical algorithm for performing ablations was arrived at in an animal model using normal liver. Limited amounts of data exist as to whether this translates to the human tumor model. METHODS: We analyzed 531 ablated lesions in 154 patients undergoing laparoscopic RFA using RITA Medical Systems Starburst XL catheter deployed to a final diameter of 2-5 cm. The first 54 patients (algorithm 1) were treated with a larger initial deployment to 3 cm and incremental advancement of the catheter to the final diameter with a 20-min ablation time for a 5-cm lesion. The subsequent 100 patients (algorithm 2) were treated with a smaller initial deployment of 2 cm, incremental advancement to the final diameter, and 14-min total ablation time for a 5-cm lesion. Lesion size was measured on 1 week postablation CT scans. Analysis was performed using the two tailed t-test. RESULTS: Ablation zones tended to be larger with the second method. On 1 week postablation CT scans, mean +/- SEM lesion sizes created using the first and second algorithms were 3.7 +/- 0.1 cm vs 4.0 +/- 0.1 cm at 3 cm deployment ( p < 0.05); 4.3 +/- 0.1 cm vs 4.8 +/- 0.1 cm at 4 cm deployment ( p < 0.05), and 5.5 +/- 0.1 cm vs 5.6 +/- 0.2 cm at 5 cm deployment ( p > 0.05), respectively. The mean +/- SEM total ablation times for the first and second algorithms were 7.9 +/- 0.3 min vs 7.0 +/- 0.2 min at 3 cm deployment ( p < 0.05); 13.3 +/- 0.3 min vs 11.1 +/- 0.02 min at 4 cm deployment ( p < 0.05); and 27.8 +/- 1.2 min vs 21.4 +/- 1.2 min at 5 cm deployment ( p < 0.05), respectively. The small SEM values indicate little variation in lesion size. CONCLUSIONS: These results show that both algorithms create dependable and reproducible zones of ablation, essential for reliable tumor destruction. Algorithm 2 demonstrates that creating an initial small core of ablation with rapid coagulation of the center of the lesion allows for equivalent, if not larger, final volumes to be performed in less time. PMID- 14735343 TI - Mediastinoscope-assisted transhiatal esophagectomy for esophageal cancer. AB - BACKGROUND: Transthoracic esophagectomy (TTE) is a radical strategy for treatment of esophageal cancer, and the morbidity and mortality are high. Transhiatal esophagectomy (THE) is advantageous because it avoids thoracotomy and has a shorter surgical time, but risk of intraoperative morbidity stresses the surgeon and lymph node sampling is not possible. METHODS: Mediastinoscope-assisted transhiatal esophagectomy (MATHE) was performed in 42 patients with esophageal cancer. Patients with superficial esophageal cancer and medical risk were included. Feasibility and efficacy of this procedure are discussed by examining short- and long-term morbidity, mortality, and survival. RESULTS: With the mediastinoscope, esophagectomy was performed safely under direct vision. There was only a small amount of bleeding, and surgical time was short. Little morbidity and no deaths were recorded. CONCLUSION: MATHE is a safe and minimally invasive technique that allows direct visualization of mediastinal structures Lymph node sampling was feasible because of clear visualization of the mediastinum. PMID- 14735345 TI - Laparoscopic pancreatic surgery: current indications and surgical results. AB - BACKGROUND: Although minimally invasive surgery has achieved worldwide acceptance in various fields, laparoscopic surgery for pancreatic diseases has been reported only rarely. The purpose of this study was to evaluate the outcomes and feasibility of laparoscopic pancreatic surgery. METHODS: Fifteen patients, comprising eight men and seven women with an average age of 54 years, underwent laparoscopic pancreatic surgery. Distal pancreatectomy was indicated for solid tumors ( n = 4), cystic lesions ( n = 3), and chronic pancreatitis ( n = 2). Cystogastrostomy was performed for pseudocysts ( n = 4) and enucleation for insulinomas ( n = 2). The lesions varied in size from 1 to 9 cm (2.9 +/- 2.4 cm) and were located in the pancreatic head ( n = 2), body ( n = 3), or tail ( n = 10). For distal pancreatectomy, the splenic artery was divided and the parenchyma was transected with a linear stapler. Laparoscopic ultrasonography was used to determine the distance between the tumor and the main pancreatic duct for enucleation as well as to localize the lesion for distal pancreatectomy. Cystogastrostomy, 4.5 cm in length, was also performed with the linear stapler through the window of the lesser omentum. RESULTS: Mean operation time was 249 +/ 70 min (293 +/- 58 min in distal pancreatectomy, 185 +/- 14 min in enucleation, 204 +/- 50 min in cystogastrostomy), and mean blood loss was 138 +/- 184 g (213 +/- 227 g, 75 +/- 35 g, 38 +/- 48 g, respectively). Two distal pancreatectomies (13%) were converted to open surgery due to severe peripancreatic inflammation. There was no related mortality, but there were two cases (15%) of pancreatic fistula, one in a distal pancreatectomy case and the other in an enucleation case, and both were treated conservatively. CONCLUSIONS: Laparoscopic pancreatic surgery is safe and feasible for patients with benign tumors and cystic lesions. PMID- 14735346 TI - Can the procedure for prolapsing hemorrhoids (PPH) be done twice? Results of a porcine model. AB - BACKGROUND: The procedure for prolapsing hemorrhoids (PPH) is a new surgical method for the treatment of symptomatic hemorrhoids. In cases of recurrent prolapse, the performance of a second PPH may result in a ring of mucosa and submucosa between the two circular staple lines. In this study, we used a porcine model to assess whether PPH can be safely performed twice. METHODS: Five adult pigs underwent two PPH procedures in one session, leaving a ring of approximately 1 cm of mucosa between the two staple lines. One month later, the pigs were examined under anesthesia. The anal canal was assessed using the following four methods: (a) clinical examination, (b) evaluation of mucosal blood perfusion at different levels of the anal canal via a laser Doppler flow detector, (c) measurement of concentrations of hydroxyproline and collagen to check for fibrosis, and (d) histopathological examination. RESULTS: At the completion of the study period, all five pigs showed no clinical evidence of anorectal dysfunction. On examination under anesthesia 1 month after surgery, there was no evidence of anal stenosis in any of the pigs. The mean mucosal blood flow between the two staple lines did not differ significantly from the flow measured proximally and distally (394 vs 363 and 339 flow units, respectively; p = NS). The collagen levels, based on hydroxyproline concentration, were 81 mcg/mg between the staple lines, compared to 82 and 79 proximally and distally, respectively ( p = NS). There was no significant difference in degree of fibrosis, as assessed histopathologically, between specimens taken from the ring between the staple lines and specimens taken from the area external to the staple lines. CONCLUSIONS: The results of this porcine model suggest that a second synchronous PPH is feasible. A controlled experience involving human subjects is required to determine the safety and usefulness of this technique in cases of metachronous application for recurrent or residual hemorrhoids. PMID- 14735350 TI - Laparoscopic relief of obstructing folded muscular cuff after transanal pull through for aganglionosis. AB - Transanal endorectal resection and colonic pull-through (TERPT) is a good technique for the management of Hirschsprung's disease. This procedure is feasible in the vast majority of patients and is associated with excellent results, early postoperative recovery, and no visible scars. We report the case of a patient who developed early postoperative severe constipation after TERPT due to unusual folding of the muscular cuff rim, which tightly narrowed the pulled-through colon. This complication was diagnosed and treated by laparoscopy. To prevent it, we recommend splitting of the aganglionic muscular cuff during TERPT. PMID- 14735348 TI - Stenting or stoma creation for patients with inoperable malignant colonic obstructions? Results of a study and cost-effectiveness analysis. AB - BACKGROUND: The aim of the present study was to compare the efficacy, safety, and cost of endoscopic palliative treatment with selfexpanding metallic stents with that of stoma creation in the management of inoperable malignant colonic obstructions. METHODS: A total of 30 patients with inoperable malignant partial obstruction (due to metastases, hemodynamic instability, or pulmonary instability) in the left colon arising from colorectal or ovarian cancer were included in the study. Fifteen were randomized to undergo palliative metallic colonic stent placement and 15 to undergo stoma creation. The efficacy and safety of the two methods was compared. A cost-effectiveness analysis was also performed, including the cost of postinterventional care. RESULTS: Stents were placed successfully in 14 of 15 patients. In one patient with obstruction of a tortuous rectosigmoid flexure colon, stenting was not possible; this patient was excluded from the study. During the follow-up period, a moderate, nonocclusive ingrowth of tumor into the stent lumen was observed in six patients; they were all treated with internal laser ablation. The cost-effectiveness analysis showed that although the stoma creation procedure was less expensive, the total difference in average costs for the two methods was 6.9% (132 Euros). CONCLUSIONS: Self-expanding metallic stent placement is a palliative alternative to colostomy for patients with inoperable malignant colonic strictures. This treatment option provides a better quality of life for the patient, without the psychological repercussions of a colostomy, and it appears to be cost-effective. PMID- 14735354 TI - Characterization of Schizosaccharomyces pombe mutants defective in vacuolar acidification and protein sorting. AB - The vacuolar H+-ATPases (V-ATPases) are ATP-dependent proton pumps responsible for acidification of intracellular compartments in eukaryotic cells. To investigate the functional roles of the V-ATPase in Schizosaccharomyces pombe, the gene vma1 encoding subunit A or vma3 encoding subunit c was disrupted. Both deletion mutants lost the capacity for vacuolar acidification in vivo, and showed sensitivity to neutral pH or high concentrations of divalent cations including Ca2+. The delivery of FM4-64 to the vacuolar membrane and accumulation of Lucifer Yellow CH were strongly inhibited in the vma1 and vma3 mutants. Moreover, deletion of the S. pombe vma1+ or vma3+ gene resulted in pleiotropic phenotypes consistent with lack of vacuolar acidification, including the missorting of vacuolar carboxypeptidase Y, abnormal vacuole morphology, and mating defects. These findings suggest that V-ATPase is essential for endocytosis, ion and pH homeostasis, and for intracellular targeting of vacuolar proteins and vacuolar biogenesis in S. pombe. PMID- 14735355 TI - Analyses of the effects of Rck2p mutants on Pbs2pDD-induced toxicity in Saccharomyces cerevisiae identify a MAP kinase docking motif, and unexpected functional inactivation due to acidic substitution of T379. AB - Rck2p is a Ser/Thr kinase that binds to, and is activated by, Hog1p. Expression of the MAP kinase kinase Pbs2pDD from a GAL1-driven plasmid hyperactivates the HOG MAP kinase pathway, and leads to cessation of growth. This toxic effect is reduced by deletion of RCK2. We studied the structural and functional basis for the role of Rck2p in mediating the growth arrest phenotype associated with overexpression of Pbs2pDD. Rck2p kinase activity is required for the effect, because Rck2p(Delta487-610), as well as full-length Rck2p, is toxic with Pbs2pDD, but kinase-defective versions of either protein with a K201R mutation are not. Thus, the C-terminal portion of Rck2p is not required provided the protein is activated by removal of the autoinhibitory domain. Relief of inhibition in Rck2p normally requires phosphorylation by Hog1p, and Rck2p contains a putative MAP kinase docking site (TILQR589R590KKVQ) in its C-terminal segment. The Rck2p double mutant R589A/R590A expressed from a centromeric plasmid did not detectably bind Hog1p-GFP and was functionally inactive in mediating the toxic effect of Pbs2pDD, equivalent to an RCK2 deletion. However, overexpression of Rck2p R589A/R590A from a multicopy plasmid restored function. In contrast, RCK2-K201R acted as a multicopy suppressor of PBS2DD, markedly reducing its toxicity. This suppressor activity required the K201R mutation, and the effect was largely lost when the docking site was mutated, suggesting suppression by inhibition of Hog1p functions. We also studied the effect of replacing the predicted T379 and established S520 phosphorylation sites in Rck2p by glutamic acid. Surprisingly, the T379E mutant markedly reduced Pbs2pDD toxicity, and toxicity was only partially rescued by S520E. Rck2 T379E was sufficiently inactive in an rck2Delta strain to allow some cells to survive PBS2DD toxicity even when overexpressed. The significance of these findings for our understanding of Rck2p function is discussed. PMID- 14735356 TI - Effects of crude extracts of Mucuna pruriens (Fabaceae) and Carica papaya (Caricaceae) against the protozoan fish parasite Ichthyophthirius multifiliis. AB - The ciliate Ichthyophthirius multifiliis is among the most pathogenic parasites of fish maintained in captivity. In the present study, the effects of the crude methanolic extract of leaves of Mucuna pruriens and the petroleum-ether extract of seeds of Carica papaya against I. multifiliis were investigated under in vivo and in vitro conditions. Goldfish (Carassius auratus auratus) infected with the parasites were immersed for 72 h in baths with M. pruriens extract, and for 96 h in baths with C. papaya extract. There was a 90% reduction in numbers of I. multifiliis on fish after treatment in baths of each plant extract at 200 mg l(-1 )compared to untreated controls. Consequently, parasite-induced fish mortality was reduced significantly. A complete interruption of trophont recruitment was achieved by immersion in the M. pruriens extract. In vitro tests led to a 100% mortality of I. multifiliis in 150 mg/l M. pruriens extract, and in 200 mg/l of C. papaya extract after 6 h. Although the active constituents of the medicinal plant extracts are still unknown, we have demonstrated that they have potential for effective control of I. multifiliis. PMID- 14735357 TI - The first ultrastructural description of the Haswell cells in Temnocephalidae (Platyhelminthes, Temnocephalida), with insights into their function. AB - This paper provides the first description of the Haswell cells at the ultrastructural level, as well as giving an insight into their function. Two species of Temnocephalidae were studied, Temnocephala iheringi and T. haswelli. Haswell cells are identical in both species, and their structure indicates that they have a secretory function. They are highly interdigitated with parenchymal cells and are usually joined to them by cup-like desmosomes. Nuclei are irregular, with a honeycomb structure and perichromatin granules. The most prominent organelle is granular endoplasmic reticulum, which is typically arranged in concentric rings that usually encircle a conspicuous Golgi complex. Secretion bodies are secreted via projections of the Haswell cells that reach the surface in the anterior portion of the body and in the tentacles. Distinct pores with a size and distribution consistent with the TEM observations were seen under SEM in these regions. PMID- 14735358 TI - Long-term in vitro cultivation of Echinococcus multilocularis metacestodes under axenic conditions. AB - We report here on the development of an in vitro system for the long-term cultivation of Echinococcus multilocularis larvae under axenic conditions. In the absence of feeder cells from the host, long-term survival of the parasite depended strictly on low oxygen conditions and the presence of reducing agents in the medium. Host serum supported survival of the parasite but the growth of metacestode vesicles and differentiation towards the protoscolex stage only occurred in the presence of culture medium that was preconditioned by hepatoma cells or several other immortal cell lines. On the basis of this in vitro system, future analyses on the identification of host-derived growth factors for E. multilocularis will be greatly facilitated. PMID- 14735359 TI - Caffey disease in a 6-month-old girl. PMID- 14735361 TI - Pathology of the pancreas and other organs in the diabetic LEW.1AR1/Ztm- iddm rat, a new model of spontaneous insulin-dependent diabetes mellitus. AB - We studied the histo- and immunopathology of the endocrine and exocrine pancreas and a number of other organs in a new insulin-dependent diabetes mellitus (IDDM) rat model (LEW.1AR1/Ztm- iddm rat). The pancreas of the acutely diabetic animals showed an inflammatory infiltrate, involving all islets and ducts. The islet infiltrate was composed mainly of ED1-positive macrophages and T lymphocytes, comprising a large number of CD8(+) lymphocytes and a few CD4(+) lymphocytes. In addition, the islets displayed apoptotic cells, characterized by condensation and fragmentation of nuclear chromatin. These cells were identified as beta cells by insulin immunostaining. Other endocrine and exocrine glands, including adrenals and thyroid, as well as salivary and submandibular glands, were unaffected. Organs from the digestive tract or systemic circulatory system, including small intestine, liver, heart, and lung also showed no involvement. The kidney was intact in acutely diabetic rats. However, 6 months after diabetes manifestation, pathological changes compatible with a diabetic nephropathy had developed, affecting both the glomerula and the proximal tubular segments. It was concluded that the autoimmune process in this new IDDM rat model is restricted to the endocrine pancreas and leads to apoptotic beta cell destruction. PMID- 14735360 TI - Gastrointestinal stromal tumours and their response to treatment with the tyrosine kinase inhibitor imatinib. AB - Gastrointestinal stromal tumours (GISTs), the most common mesenchymal tumours of the digestive tract, are largely resistant to chemo- and radiotherapy. They are currently defined by their overexpression of the KIT receptor tyrosine kinase (CD117), a member of the family of receptor tyrosine kinases (RTKs), and exhibit KIT mutations in more than 85% of cases. Additionally, in more than one-third of KIT wild-type GISTs, mutations of platelet-derived growth factor receptor alpha (PDGF-R alpha), which also belongs to the family of RTKs, were recently found. Since these data indicate that uncontrolled RTK signalling may be implicated in the pathogenesis of GISTs, RTKs and the activated downstream signalling cascades are attractive targets in the therapy of these tumours. Imatinib is a small molecule inhibitor that selectively blocks the activity of the PDGF-R, ABL and KIT receptor tyrosine kinases by competitive binding to the adenosine triphosphate binding site of their catalytic domains. We herein review the molecular pathological, preclinical and clinical data that identify imatinib as a valuable new agent in the treatment of GISTs. PMID- 14735362 TI - Molecular analysis of the 5'-upstream region of a gibberellin-inducible cytosolic glutamine synthetase gene (GS1b) expressed in pine vascular tissue. AB - The promoter region of the cytosolic glutamine synthetase GS1b gene was isolated from the conifer Pinus sylvestris L. (Scots pine). The 1,171-bp stretch of sequence lying immediately upstream of the transcriptional start site was sufficient to drive the expression of a reporter gene in a manner consistent with the expression pattern of the native GS1b gene. Computer analysis of putative cis elements in this promoter region revealed the presence of an AT box, an AC motif similar to those found in other genes expressed in the vascular tissue, and a gibberellin (GA)-responsive element. Consistent with the latter finding, GS1b gene expression was induced by exogenously supplied gibberellic acid (GA3) in germinating pine embryos and pine seedlings. In order to examine if the putative GA-response element found in the GS1b promoter could function in the regulation of GS1b expression, a series of deletions of the upstream gene region were fused to the uidA reporter gene, and transient expression analyzed either in untreated or in GA3-treated pine (Pinus pinaster Ait.) protoplasts. Deletion analysis revealed that sequences containing the GA-responsive element, located between 1005 and -724 bp were essential for the increased promoter activity observed in response to GA3. Furthermore, electrophoretic-mobility-shift assays showed that pine nuclear proteins bind to a 22-bp sequence that contains the GA-response element, located between -768 and -747 bp relative to the transcription start site. PMID- 14735363 TI - Chest wall kinematics and respiratory muscle coordinated action during hypercapnia in healthy males. AB - The present study was designed to verify whether during hypercapnic stimulation, as we had previously found during exercise or walking, the partitioning of the respiratory motor output is equally distributed to the muscles of chest wall compartments to assist diaphragm function. We studied chest wall kinematics and respiratory muscle recruitment in seven healthy men during rebreathing of a hypercapnic-hyperoxic gas mixture (CO(2) RT). Data were compared with those previously obtained during either cycling exercise or walking. The chest wall volume ( Vcw), assessed by optoelectronic plethysmography (OEP), was modeled as the sum of the volumes of the lung-apposed rib cage ( Vrc,p), diaphragm-apposed rib cage ( Vrc,a) and abdomen ( Vab). Esophageal ( Pes), gastric ( Pga) and transdiaphragmatic ( Pdi= Pga- Pes) pressures were simultaneously recorded. Velocity of shortening ( V') and power ( W'= Px V') of the diaphragm ( W'di), rib cage muscles ( W'rcm) and abdominal muscles ( W'abm) were also calculated. During CO(2) RT the progressive increase in end-inspiratory Vcw resulted from an increase in both end-inspiratory Vrc,p and Vrc,a, while the progressive decrease in end-expiratory Vcw was entirely due to the decrease in end-expiratory Vab. The increase in Vrc,p was proportionally slightly greater than that in Vrc,a. The end inspiratory increase and end-expiratory decrease in Vcw were accounted for by inspiratory rib cage (RCM,i) and abdominal (ABM) muscle recruitment, respectively. W'di, W'rcm and W'abm progressively increased. However, while most of W'di was expressed in terms of velocity of shortening, most of W'rcm and W'abm was expressed as force or pressure. A comparison of CO(2) results with data obtained during exercise revealed: (1). a gradual vs. an immediate response, (2). a similar decrease in Vab,e and Pabm, (3). an apparent lack of any difference in ABM recruitment, (4). less gradual ABM relaxation, (5). no drop in Pdi but a similar Wdi change and decrease in pressure-to-velocity ratio of the diaphragm. We have found that in healthy humans: (1). the increased motor output with hypercapnia is equally distributed between RCM and ABM to minimize transdiaphragmatic pressure and (2). data on chest wall kinematics and respiratory muscle recruitment are only partly in line with those obtained during walking or cycling exercise. PMID- 14735364 TI - Influences of age and sex on abdominal muscle and subcutaneous fat thickness. AB - This study aimed to investigate the influences of age and sex on the thickness of the rectus abdominis muscle and subcutaneous fat at the abdomen. The two tissue thickness values were determined using ultrasonography in a total of 194 young (20-29 years) and elderly (70-79 years) persons of both genders. In both sexes, the elderly groups showed significantly thinner muscle and thicker fat than the younger groups. The percentage of the mean value of subcutaneous fat thickness in the elderly group compared to that in the younger group was higher in the women (176%) than in the men (128%), but the corresponding value of muscle thickness was similar between men (68%) and women (65%). In both generations, the women had significantly thinner muscle and thicker fat than the men. The percentage of the mean value of subcutaneous fat thickness in the women compared to that in the men was higher in the elderly group (169%) than in the younger group (123%), but the corresponding value of muscle thickness was similar between the younger (80%) and elderly (77%) groups. In every group, subcutaneous fat thickness significantly correlated with waist circumference, but muscle thickness did not. The findings here indicated that: (1). aging was associated with a decrease of muscle thickness and an increase of subcutaneous fat at the abdomen in men and women, with a similar relative loss of muscle thickness in both genders and a greater increase of subcutaneous fat thickness in women than in men, and (2). subcutaneous fat rather than muscle thickness reflected waist circumference, regardless of age and gender. PMID- 14735365 TI - Simple method for the identification of oxidative fibers in skeletal muscle. AB - Skeletal muscle is composed of several different types of myofiber: slow oxidative (SO), fast glycolytic oxidative and fast glycolytic. However, the classification is usually determined by myosin heavy chain typing rather than by metabolic index. In this study, the oxidative metabolic index was investigated as a possible method of myofiber typing. Myoglobin, which is involved in oxygen transport and storage in myofibers, and mitochondria, which are the central organelles for oxidative metabolism, were studied. High levels of myoglobin and mitochondria are believed to exist in SO fibers, but the current study showed that they are considerably richer in some fast type fibers. As myofiber typing using the oxidative metabolic index is important physiologically, an attempt was made to find a simple method for this purpose. Some mitochondrial proteins have been observed to auto-fluoresce but until now this effect was too faint to detect easily. Owing to the recent advances in cooling charge-coupled device technology, such auto-fluorescence can now be used for myofiber typing, and the simple and rapid method for doing so is reported here. PMID- 14735366 TI - Computer-assisted skin videothermography is a highly sensitive quality tool in the diagnosis and monitoring of complex regional pain syndrome type I. AB - The use of thermography in the diagnosis and evaluation of complex regional pain syndrome type 1 (CRPS1) is based on the presence of temperature asymmetries between the involved area of the extremity and the corresponding area of the uninvolved extremity. The interpretation of thermographic images is, however, subjective and not validated for routine use. The objective of the present study was to develop a sensitive, specific and reproducible arithmetical model as the result of computer-assisted infrared thermography in patients with early stage CRPS1 in one hand. Eighteen patients with CRPS1 on one hand and 13 healthy volunteers were included in the study. The severity of the disease was determined by means of pain questionnaires [visual analogue scale (VAS) pain and McGill Pain Questionnaire], measurements of mobility (active range of motion) and oedema volume. Asymmetry between the involved and the uninvolved extremities was calculated by means of the asymmetry factor, the ratio and the average temperature differences. The discrimination power of the three methods was determined by the receiver-operating curve (ROC). The regression between the determined temperature distributions of both extremities was plotted. Subsequently the correlation of the data was calculated. In normal healthy individuals the asymmetry factor was 0.91 (0.01) (SD), whereas in CRPS1 patients this factor was 0.45 (0.07) (SD). The performance of the arithmetic model based on the ROC curve was excellent. The area under the curve was 0.97, the P value was <0.001, the sensitivity 92% and specificity 94%. Furthermore, the temperature asymmetry factor was correlated with the duration of the disease and VAS pain. In conclusion, in resting condition, videothermography is a reliable additive diagnostic tool of early stage CRPS1. This objective tool could be used for monitoring purposes during experimental therapeutic intervention. PMID- 14735367 TI - Recurrent retinal vein occlusion after playing a wind instrument. AB - PURPOSE: To present a complication of repeated Valsalva maneuver. METHODS: Case report with serial intravenous fluorescein angiography and comprehensive workup for coagulopathies. RESULTS: After playing strenuous musical pieces, a solo trombonist suffered within 1 year three attacks of visual loss in the right eye; he had two attacks of inferior hemi-central retinal vein occlusion 2 months apart, followed by central retinal vein occlusion 10 months later. Final best spectacle-corrected visual acuity was 6/60. He ended his professional career, and no further attacks occurred in the following 10 years. CONCLUSION: Retinal vein occlusion with irreversible visual loss can occur after playing a strenuous piece of music on a high-resistance instrument. PMID- 14735368 TI - Spontaneous reactivation of a silent telomeric transgene in a human cell line. AB - Subtelomeric reporter genes in human cells are silenced in a telomere length dependent manner. Here we show that a subtelomeric reporter gene is expressed in only a subpopulation of cells within a clone and that this heterogeneity is generated by switching between expression states. We observed frequent reversion from the silenced state back to active expression. This process was more prominent for subtelomeric transgenes; however, we also observed cases of spontaneous reversion in control clones bearing the reporter at an internal site. We additionally show that treatment of these cells with 5-bromodeoxyuridine results in strong activation of the transgene. Although similar findings have been reported previously in mouse cells, this is, to our knowledge, the first direct observation of ongoing fluctuations in transcription in clonal populations of human cells. Our results suggest that this mechanism, as opposed to progressive silencing or a delayed fixing of expression states, accounts for the variegation in expression observed for subtelomeric transgenes in human cells. These data imply that telomere shortening during human aging could lead to stochastic activation of subtelomeric genes. PMID- 14735369 TI - The scrambled actin I gene in Uroleptus pisces. AB - The micronuclear gene encoding actin I in Uroleptus pisces occurs in two segments. Segment I contains 638 bp divided into six macronuclear destined subsegments, or MDSs, by five internal eliminated segments, or IESs. The MDSs in segment 1 are in the scrambled disorder, 1-2-4-8-6-15, with MDSs 8 and 6 inverted. Segment II contains 2452 bp divided into ten MDSs by nine IESs in the scrambled disorder, 3-5-7-10-13-12-9-14-16-11, with MDSs 12, 9, and 11 inverted. Extensive attempts by polymerase chain reaction to connect the two segments failed. We conclude that the two segments are separated by a very long IES or are in different loci. The pattern of the 16 scrambled MDSs is entirely different from the scrambled pattern observed for the actin I gene in six other stichotrichs. We conclude that the actin I gene became scrambled on two separate occasions during stichotrich evolution: once in the lineage leading to the group of six stichotrichs, which includes, among others, Sterkiella species and Stylonychia lemnae, and once in the lineage leading to Uroleptus pisces. Repeated sequence pairs (pointers) of three to 14 bases at the ends of MDSs appear to be essential for correct splicing of MDSs during macronuclear development. However, the micronuclear actin gene also contains 40 matches of eight or more bases between IESs and MDSs that do not function as pointers. To prevent these ectopic repeats from causing improper processing of the micronuclear gene appears to demand a template of DNA or RNA from the old macronucleus to guide splicing of MDSs in the orthodox order. PMID- 14735370 TI - Microirradiation of cells with energetic heavy ions. AB - The ion microprobe SNAKE at the Munich 14 MV tandem accelerator achieves beam focussing by a superconducting quadrupole doublet and can make use of a broad range of ions and ion energies, from 20 MeV protons to 200 MeV gold ions. Because of these properties, SNAKE is particularly attractive for biological microbeam experiments. Here we describe the adaptation of SNAKE for microirradiation of cell samples. This includes enlarging of the focal distance in order to adjust the focal plane to the specimen stage of a microscope, construction of a beam exit window in a flexible nozzle and of a suitable cell containment, as well as development of procedures for on-line focussing of the beam, preparation of single ions and scanning by electrostatic deflection of the beam. When irradiating with single 100 MeV (16)O ions, the adapted set-up permits an irradiation accuracy of 0.91 microm (full width at half maximum) in the x direction and 1.60 microm in the y-direction, as demonstrated by retrospective track etching of polycarbonate foils. Accumulation of the repair protein Rad51, as detected by immunofluorescence, was used as a biological track detector after irradiation of HeLa cells with geometric patterns of counted ions. Observed patterns of fluorescence foci agreed reasonably well with irradiation patterns, indicating successful adaptation of SNAKE. In spite of single ion irradiation, we frequently observed split fluorescence foci which might be explained by small scale chromatin movements. PMID- 14735371 TI - The dynamics of precursor cells in the olfactory epithelium of juvenile and adult guinea pigs. AB - The dynamics of precursor cells in the olfactory epithelium of juvenile and adult guinea pigs were examined by immunohistochemical double staining using bromodeoxyuridine (BrdU), the neural cell adhesion molecule (N-CAM) and the protein gene product 9.5 (PGP9.5). Expression of apoptotic cells in the olfactory epithelium with the use of the TdT-mediated dUTP biotin nick end labeling (TUNEL) method was also observed. BrdU was given to healthy guinea pigs at the ages of 2 weeks and 6 months old. Tissue specimens were serially collected 1 h to 28 days after administration. BrdU-labeled cells were seen above the basal cell layer after 1 h and migrated to the middle layer of the olfactory epithelium, after 1 day in juveniles and 5 days in adults with expression of N-CAM. PGP9.5 was observed in BrdU-labeled cells after 5 days in juvenile guinea pigs and 7 days in adult. At 14 days after administration, BrdU-labeled cells in the epithelium appeared to decrease. However, a few of these cells were recognized above the basal cell layer after 28 days. The number and location of TUNEL-positive cells did not significantly differ between the juvenile and adult olfactory epithelium. Therefore, we conclude that the division speed from stem cells in juveniles is faster than that in adults, and apoptosis is unaffected by aging in the normal olfactory epithelium. PMID- 14735373 TI - Abnormal uterine bleeding as a presentation of metastatic breast disease in a patient with advanced breast cancer on tamoxifen therapy. AB - INTRODUCTION: Extra pelvic tumours rarely metastasise to the female genital tract. Typically it is the ovaries that are involved and the primary tumours include the breast and the gastrointestinal tract. Metastatic disease involving the uterus is rare. CASE REPORT: A case of metastatic breast carcinoma involving both the uterine corpus and cervix is described. The patient was known to have advanced breast carcinoma and was on tamoxifen therapy when she presented with postmenopausal bleeding. Histological assessment of the hysterectomy specimen including immunohistochemistry staining confirmed the breast carcinoma as the primary. CONCLUSION: Unusual uterine bleeding in a patient with breast cancer, should alert the gynaecologist to the possibility of metastatic breast disease regardless of tamoxifen treatment. PMID- 14735372 TI - Coagulation inhibitors in preeclamptic pregnant women. AB - OBJECTIVE: Our objective was to detect clinical evaluation of coagulation inhibitors in preeclamptic and normotensive pregnant women and to determine their important role in pathogenesis of preeclampsia. METHODS: A total of 20 mild, 20 severe preeclamptic and 45 normotensive pregnant women were included in this study. The plasma value of antithrombin III (AT-III) activity, proteins C and S activity, PT, PTT, fibrinogen and platelet counts were determined. RESULTS: The values AT-III were lower in women with severe preeclampsia than in controls (p<0.05). In all groups, there was no significantly difference in the concentration of protein C activity, protein S and fibrinogen (p>0.05). The plasma thrombocyte counts were significantly lower in severe preeclamptic women than in normotensive women (p<0.05). There was no significant difference in the prothrombin time value in all groups, but a significantly difference with regard to partial thromboplastin time between severe preeclamptic and the control group (p<0.0001). It was longer than the control. CONCLUSION: The markers of hemostasis activation such as protein S, protein C activity together with fibrinogen levels are not useful tools but the reduction of AT-III and platelet counts would seem useful in different pathological situations in pregnancy to predict and monitor the severity of the condition. PMID- 14735374 TI - Primary malignant mixed mullerian tumor of the vagina immunohistochemically confirmed. AB - CASE REPORT: We report the case of a 74-year old woman who presented with an ulcerated mass of the vagina. Histology of the tumor showed malignant mixed mullerian tumor (MMMT) with squamous and spindle cell stromal components, associated with high-grade vaginal intraepithelial neoplasia (VaIN 3). Immunohistochemical study of the neoplasm revealed that the malignant stromal tumor was a high-grade leiomyosarcoma. Despite the multimodal therapeutic approach, the patient died within 11 months. CONCLUSION: The vagina is a rare site of presentation of primary MMMTs and the present case is the second one immunohistochemically confirmed. PMID- 14735375 TI - Severe vasovagal attack: an unusual cause of abruptio placentae. AB - CASE REPORT: Abruptio placentae occurred in a 29 years old woman following a vasovagal episode. Her medical history was free from all the commonly accepted risk factors for abruption. DISCUSSION: We speculate that the restoration of placental blood flow caused a sudden increase in uteroplacental blood pressure, and induces rupture of some vessels, causing a progressively growing retroplacental hematoma and placental detachment. PMID- 14735376 TI - Tuberculosis-any new insights? PMID- 14735377 TI - BKV-DNA and JCV-DNA in CSF of patients with suspected meningitis or encephalitis. AB - BACKGROUND: Few studies have looked for the polyoma viruses JC or BK virus in the central nervous system (CNS) of patients without neurological symptoms or with neurological symptoms other than progressive multifocal leukoencephalopathy (PML). PCR-microplate hybridization method was employed for the detection of BKV DNA or JCV-DNA in cerebrospinal fluid (CSF) specimens from patients with suspected meningitis or encephalitis. MATERIALS AND METHODS: A total of 181 CSF specimens from 151 patients with suspected meningitis or encephalitis was examined for BKV or JCV using PCR-microplate hybridization method. None of the patients had (clinically diagnosed) PML. A control group consisting of 20 CSF specimens from normal subject was also included. RESULTS: BKV DNA was found in five out of 131 (3.8%) and JCV DNA in two out of 131 (1.5%) of the patients with suspected meningitis or encephalitis by PCR ELISA. BKV or JCV DNA was not detected in CSF samples of any of 19 HIV positive patients. BKV and JCV DNAs were detected respectively in two CSF samples in which Mycobacterium tuberculosis (TB) PCR was also positive. Another patient who was positive for JCV PCR died with a diagnosis of cerebral lymphoma. Among the BK virus infected patients there was a patient with a previous history of hemolytic uremia and acute renal failure. Neither BKV nor JCV DNA was found in any of the 20 CSF samples from normal patients undergoing lumbar puncture for myelography as a part of an investigation of lower back pain. CONCLUSION: These results suggest that BK virus may be associated with neurological diseases either in immunocompetent or immunocompromised patients. Detection of BKV and JCV DNA in the CSF of the patients suspected to have either meningitis or encephalitis suggests that these viruses may have an etiological role. Thus, diagnostic tests for BK and JC viruses should be included in the investigative program for meningitis or encephalitis patients. PMID- 14735378 TI - Reduction of serum melatonin levels in HIV-1-infected individuals' parallel disease progression: correlation with serum interleukin-12 levels. AB - BACKGROUND: During the natural history of human immunodeficiency virus type I (HIV-1) infection, an impairment of interleukin-12 (IL-12) production precedes a switch from a T-helper 1 (Th1) to a T-helper 2 (Th2) stage of cellular immunity. Melatonin, the main hormone produced by the pineal gland, seems to promote a Th1 response by increasing the production of IL-12 in vitro. The aim of this study was to measure and correlate serum levels of melatonin and IL-12 in a cohort of HIV-1-infected individuals. PATIENTS AND METHODS: 77 anti-HIV-1-positive subjects were enrolled: 20 were in CDC stage A, 25 in CDC stage B and 32 in CDC stage C. 30 healthy HIV-1-seronegative subjects were recruited as controls. IL-12 and melatonin concentrations were quantitated in serum samples. RESULTS: Mean levels of serum melatonin were significantly lower in HIV-1-infected individuals in comparison with controls (p < 0.001). Within the HIV-1-seropositive group, mean melatonin and IL-12 concentrations were significantly lower in patients in CDC stage C, as compared with patients in CDC stages B and A (p < 0.01). CONCLUSION: During the natural history of HIV-1 disease, serum melatonin levels are progressively reduced. This reduction may be related to the impairment of Th1 immunoresponses. PMID- 14735379 TI - Oral lichen planus in patients with chronic liver diseases. AB - BACKGROUND: The possible increased risk of oral lichen planus (OLP) in patients with chronic liver diseases is a controversial issue. PATIENTS AND METHODS: We investigated a total of 156 outpatients with known liver diseases for occurrence of OLP. Hepatitis C virus (HCV) infection was proven for 117 patients (75%) and hepatitis B virus (HBV) infection for 16 patients (10.3%). Four patients were coinfected with HCV and HBV. An alcohol-derived hepatic cirrhosis was found in 23 patients (14.7%). Three patients had a biliary cirrhosis and one had a primary sclerosing cholangitis. Patients suspected of having OLP were transferred to the Department of Oral and Maxillofacial Surgery where excisional biopsies were taken. RESULTS: Eight patients had a white alteration of the oral mucosa. OLP was suspected in five of them and was confirmed histologically in four. The alteration of the fifth patient disappeared during continued interferon therapy. CONCLUSION: Our results show that there is no increased prevalence of OLP manifestation in patients suffering from HCV infection. PMID- 14735380 TI - Tuberculous meningitis in adults: review of 61 cases. AB - BACKGROUND: Tuberculosis is still a major cause of serious illness in developing countries. The objective of this study was to assess the clinical, laboratory, radiological and prognostic features of tuberculous meningitis (TBM) in immuncompetent adult patients. PATIENTS AND METHODS: Sixty-one patients with confirmed or presumed TBM seen over a 12-year period at the Neurology Department of Bakirkoy Research and Training Hospital for Psychiatric and Neurological Disease, Istanbul, Turkey, were analyzed retrospectively. Patients were grouped according to the severity of meningitis on admission (stages I, II and III). They were also divided into two groups according to the presence of paradoxical response (progressive increase of lymphocytes or increase of polymorphonuclear cells instead of lymphocytes) in CSF samples. A combination of five antituberculosis drugs was used in the 1st month of treatment. Patients received antituberculosis therapy for at least 12 months. The outcome of the patients was defined on the basis of the 12-month Barthel index (BI) score (BI < 12 poor; BI >/= 12 good). For statistical analysis death was included in the poor outcome group. RESULTS: Paradoxical response in CSF findings was seen in 20 patients. There was no statistically significant difference between the outcome of the patients with and without CSF paradoxical response; however, new tuberculomas developed more frequently in the first group (p < 0.05). The overall mortality was 27.8%. Stage of disease was found to be independently associated with the 12 month outcome (OR 7.2; 95% CI 1.7-30.3, p = 0.007). CONCLUSION: In developing countries such as Turkey, tuberculosis is still an important public health issue. Early suspicion and appropriate long-term antituberculosis therapy together with corticosteroids may reduce mortality and morbidity in TBM patients. PMID- 14735381 TI - Meningococcal disease in catalonia 1 year after mass vaccination campaign with meningococcal group C polysaccharide vaccine. AB - BACKGROUND: The aim of this study was to investigate the clinical and epidemiological characteristics of meningococcal disease in Catalonia (Spain) after vaccination with the polysaccharide vaccine. PATIENTS AND METHODS: Cases were collected by the Statutory Diseases Reporting System. RESULTS: 176 cases were reported, an overall incidence of 2.9/100,000 persons/year. 60% of cases occurred during winter and spring. The case fatality rate was 6.3%. The highest age incidence was in children under 2 years of age (48/100,000 persons/year). Comparison of the cases detected by the Statutory Diseases Reporting System with those obtained by the Microbiological Reporting System shows that meningococcal disease surveillance in Catalonia was relatively complete (95.7%), with a positive predictive value of 66.3%. 115 cases (65%) were culture-confirmed with a rate of 1.9/100,000 persons/year. 86 (75%) cases were due to Neisseria meningitidis serogroup B and 21 to serogroup C (18%). CONCLUSION: Although infections due to serogroup C have decreased after mass vaccination with the polysaccharide vaccine, it is likely that the number of infections will decrease further with the conjugate meningococcal group C vaccine. PMID- 14735382 TI - Prospective survey of antibiotic utilization in pediatric hospitalized patients to identify targets for improvement of prescription. AB - BACKGROUND: The rise in the use of antibiotics has resulted in increasing health care costs and the emergence of resistant bacteria. Little is known about the general misuse of antibiotics in hospitalized children. We evaluated the utilization of antibiotics in a pediatric teaching hospital aiming to identify targets for improvement of prescription. PATIENTS AND METHODS: Clinical, radiological, laboratory and treatment data of patients hospitalized in a pediatric medical and a pediatric surgery ward were prospectively collected during a 6-week period. A subsequent review of the collected data by a pediatric infectious diseases specialist, taking into consideration existing in-house treatment guidelines, was carried out. RESULTS: A total of 125 (36%) of 349 patients was prescribed 246 antibiotics. The median length of hospital stay for children prescribed antibiotics was 5 days (range, 2-30 days) and for those not prescribed 3 days (1-32 days; p < 0.001). Of 154 patients in the medical ward, 64 (42%) received antibiotics, compared to 61 (31%) of 195 patients in the surgical ward (p < 0.05). Empirical prescriptions were more frequent than prophylactic ones, which were more frequent than therapeutic prescriptions (136 [55%] vs 94 [38%] vs 16 [7%]; p < 0.001). Overall, 85% of the prescriptions were considered justified. The rates of inappropriate prescriptions were similar in the medical and surgical ward, and higher for therapeutic (19%) or prophylactic treatment (18%) than for empirical treatment (12%). Higher inappropriate prescription rates were noted for macrolides than for co-trimoxazole and beta-lactams (50% vs 18% and 15%, respectively; p < 0.05). CONCLUSION: Efforts need to be undertaken towards continuous education of medical staff on judicious antibiotic use, as well as ensuring compliance with existing guidelines. Improvement in the availability of rapid diagnostic methods to discern viral from bacterial infections may help reduce the numbers of empiric therapies in favor of pathogen targeted therapeutic treatments. PMID- 14735383 TI - Solitary and multiple erythema migrans in children: comparison of demographic, clinical and laboratory findings. AB - BACKGROUND: Data on European children with erythema migrans (EM) are limited. PATIENTS AND METHODS: 553 patients, 333 with solitary and 220 with multiple EM, diagnosed between 1996 and 2000, were included in the prospective study. Demographic, clinical and laboratory data including borrelial serum immunofluorescence assay antibody titers and Borrelia burgdorferi sensu lato blood culture results were obtained; findings in solitary and multiple EM were compared. RESULTS: Comparison revealed that children with multiple EM were younger (4.5 vs 6.5 years; p = 0.0000), less often reported a tick bite at the site of later skin lesion (25% vs 46%; p = 0.0000), had a longer incubation period (22 vs 13 days; p = 0.0028), more frequently presented with a ringlike lesion (99% vs 86%; p = 0.0000), less often reported associated local (15% vs 41%; p = 0.0000) but not systemic symptoms (28% vs 26%, p = 0.6913), more frequently had abnormal findings on physical examination (35% vs 26%; p = 0.0264), and a higher frequency of laboratory abnormalities including the presence of borrelial serum antibodies as well as B. burgdorferi sensu lato isolated from blood (12% vs 6%; p = 0.0267); younger age and male sex were identified as risk factors for the isolation of Borrelia. 40/44 isolates were Borrelia afzelii. CONCLUSION: Analysis of a large group of European children with solitary and multiple EM revealed several demographic, clinical and laboratory differences between the two groups. PMID- 14735384 TI - Chlamydia pneumoniae carriage and infection in hospitalized children with respiratory tract diseases. AB - BACKGROUND: The importance of Chlamydia pneumoniae respiratory tract infection in childhood is under discussion. PATIENTS AND METHODS: 798 hospitalized children with respiratory tract diseases were prospectively studied during a 2-year period by polymerase chain reaction and enzyme immunoassay (PCR-EIA) detection from throat swabs. Paired serum samples were used to screen for Chlamydia antibodies. RESULTS: C. pneumoniae was detected by PCR-EIA in 74 children. Prevalence was 11% in lower and 4% in upper respiratory tract disease (p = 0.049) without age dependency. From November to February prevalence was elevated (42/277 vs. 32/521; p < 0.001). Using serology, prevalence of acute Chlamydia infection increased with age (p < 0.001) and the number of coinfections (p < 0.001), without seasonal variation. CONCLUSION: Characteristics of C. pneumoniae carriage in the respiratory tract in childhood differ from those in systemic infection. PMID- 14735385 TI - Fever of unknown origin in Turkey. AB - BACKGROUND: The etiology of fever of unknown origin (FUO) includes primarily infectious, collagen-vascular and neoplastic diseases. The distribution of the disorders causing FUO may differ according to the geographic area and the socioeconomical status of the country. Moreover, the developments in radiographic and microbiologic methods have changed the spectrum of diseases causing FUO. MATERIALS AND METHODS: We reviewed 117 cases that fulfilled the criteria of FUO followed in our department during the period 1984 to 2001. RESULTS: The etiology of FUO was infectious diseases in 34% of the patients, collagen-vascular diseases in 23%, neoplasms in 19% and miscellaneous diseases in 10%. In 14% of the cases the etiology could not be found. The three leading diseases were tuberculosis (24%), lymphomas (19%) and adult-onset Still's disease (11%). Tuberculosis was found to be a more common cause of FUO than reported in studies in developed countries. Invasive procedures helped to establish the diagnosis in 50 out of 92 patients (43%). As a final diagnostic procedure, laparotomy aided the establishment of a diagnosis in 15 out of 20 patients (75%). CONCLUSION: Although the relative rate of infectious disease as etiologic category is less commonly encountered, infectious disease, especially tuberculosis, remains a common cause of FUO. Although several diseases may lead to FUO, lymphomas, adult-onset Still's disease and particularly tuberculosis should be considered in the differential diagnosis of a patient admitted with FUO. PMID- 14735386 TI - Catheter-related bacteremia caused by Agrobacterium radiobacter in a cancer patient: case report and literature review. AB - Agrobacteria are a group of phytopathogenic organisms widely distributed in soil; they are now recognized as rare human pathogens affecting mostly immunocompromised hosts. We report a case of catheter-related bacteremia due to Agrobacterium radiobacter in a neutropenic patient and describe the clinical presentations, treatment strategies and outcome of Agrobacterium infections based on our experience and a literature review. The antimicrobial susceptibility patterns of these organisms appear to be quite variable and collective susceptibility data derived from this and previous reports are provided. PMID- 14735387 TI - Meningitis due to multiple-resistant penicillin- and cefotaxime-intermediate Streptococcus pneumoniae in a German child after bone marrow transplantation. AB - The incidence of infection with penicillin-non-susceptible Streptococcus pneumoniae is increasing rapidly worldwide. Spain and France are highly affected, whereas the level of penicillin resistance in Germany, Italy, The Netherlands and Scandinavia is low. We report a lethal episode of meningitis due to penicillin- and cefotaxime-intermediate S. pneumoniae in a 7-year-old, allogene bone marrow transplanted German boy, 5 weeks after a holiday in Spain. Three days prior to the infection the patient showed good performance status. He was in complete remission without signs of graft-versus-host disease (GVHD). He died on day 341 post bone marrow transplant (BMT), 2 days after the onset of meningitis. Penicillin-non-susceptible S. pneumoniae should be regarded as a potential infectious agent even in countries with a low prevalence of resistance. PMID- 14735388 TI - Ciprofloxacin-induced toxic epidermal necrolysis in a patient with systemic lupus erythematosus. AB - Toxic epidermal necrolysis (TEN), or Lyell's syndrome, is a fulminant bullous dermatitis. TEN is often a drug-induced reaction and virtually any drug class appears capable of provoking it. We report here a case of TEN after administration of ciprofloxacin. Systemic lupus erythematosus (SLE) was suspected as a possible etiologic or modifying cofactor in TEN in this case. PMID- 14735389 TI - A case of Opisthorchis felineus infestation in a pilot from Greece. AB - We describe the case of a 28-year-old man from Greece with Opistorchis felineus infestation. The patient presented with intense abdominal pain, bilious emesis and eosinophilia. He probably acquired the infection overseas, since he was a commercial airline pilot who used to fly to endemic areas and to consume raw or undercooked fish. He was successfully treated with praziquantel administered in divided doses over a single day. Opisthorchiasis is common to eastern Europe and areas of the former Soviet Union, but extremely rare in Greece. Medical personnel should be cognizant of this parasitic infection, since world travel can spread it to areas of the world unaccustomed to it. PMID- 14735390 TI - A comparison of buttress drumming by male chimpanzees from two populations. AB - Wild chimpanzees ( Pan troglodytes) produce low-frequency sounds by hitting the buttresses and/or trunks of trees. This "buttress drumming" occurs in discrete bouts that may be integrated into the phrase sequence of the chimpanzee's long distance vocalization, the "pant hoot." The aim of this study was to investigate whether regional variation exists in the drumming behavior of male chimpanzees from Kibale National Park (Kanyawara community), Uganda, and Tai National Park, Ivory Coast. Recordings were made during a 6-month field season at Tai in 1990, and a 12-month field season at Kanyawara in 1996-1997. Acoustic analysis revealed the following: (1) Kanyawara males drummed significantly less frequently in conjunction with a pant hoot or hoot than did Tai males; (2) drumming bouts by Kanyawara males included significantly fewer beats, and were significantly shorter in duration, than those of Tai males; these differences disappeared when only those bouts produced in conjunction with a call were compared; (3) when Kanyawara chimpanzees did call and drum together, they tended to integrate drumming into the vocalization at a later point than did Tai males; and (4) individual differences in the temporal patterning of drumming bouts were not apparent for Kanyawara males, whereas a previous analysis revealed individual differences among Tai males. PMID- 14735391 TI - Field testing a global positioning system (GPS) collar on a Japanese monkey: reliability of automatic GPS positioning in a Japanese forest. AB - A global positioning system (GPS) collar recorded the locations of an adult female Japanese macaque over a 9-day period in a habitat with mixed suburban and rural land-uses in Chiba Prefecture, Japan. The GPS device acquired positions even in forested areas. The GPS data located the female mostly in forested areas, although the female had ranged through a habitat with inter-mingled fields, orchards, quarries, and residential areas. However, the GPS position acquisition rate was low compared to studies carried out on North American mammals. The GPS fixed a position in 20% of positioning attempts. When the collared female was tracked by radio-telemetry, almost all failures of the GPS to fix a position occurred in forest. PMID- 14735392 TI - Diet and activity in black howler monkeys ( Alouatta pigra) in southern Belize: does degree of frugivory influence activity level? AB - This study reports on the diet and activity budgets of Central American black howling monkeys ( Alouatta pigra) at Monkey River, Belize. This is a previously unstudied population, close to the southern boundary of the species range, and it provides comparative data on A. pigra from a new study site. Both diet and activity are within the ranges reported for other A.pigra sites and for mantled howlers ( A. palliata). No age-sex differences could be discerned in either diet or activity, though monthly variation was apparent. The monkeys switch from consuming leaves 86% of the time in January to March to consuming 67% fruit in April to July. This difference was statistically significant, and provided the opportunity to compare activity levels of the monkeys over two dietary periods, one characterized primarily by folivory, the other by frugivory. Howlers are often seen as a relatively inactive species, something that is associated with a low quality, folivorous diet. However, A. pigra have been described as being as frugivorous as possible and as folivorous as necessary. Yet, despite the opportunistic consumption of large quantities of high-energy foods, A. pigra has been observed as conforming to the howler lifestyle, resting as much as 80% of the day. The data in this paper support both of these reports. Black howlers at Monkey River Belize are typically inactive, maintaining high levels of inactivity even during months characterized by frugivory, suggesting that diet is more flexible and varied than is behavior and calling into question the assumption that howler inactivity is due to the digestion of large quantities of leaves. PMID- 14735393 TI - Microbial characterisation of activated sludge in jet-loop bioreactors treating winery wastewaters. AB - Jet-loop reactors (JLR) used as biological waste treatment processes introduce an additional selective pressure on the natural microbial flora of the incoming effluent. Several high-performing microbial inocula were tested for winery wastewater treatment and the microbial composition was analysed. A microbial consortium was enriched and selected for use with a new type of aerobic JLR. The reactor was operated continuously for more than 1 year using winery wastewaters collected in different seasons. Chemical oxygen demand (COD) removal efficiency was on average greater than 80%, with retention times of 0.8-1 day. Microbial populations were sampled for characterisation after 6 months and at the end of the study. Isolates were identified at genus and/or species level. Almost all isolates belonged to the genera Pseudomonas and Bacillus. Saccharomyces cerevisiae was also found but no filamentous fungi. These results show that a highly adapted population develops in JLRs treating winery effluents as compared to other bioreactors. Aerobic JLRs impose a stringent selective criterion on the composition of the microbial biomass. PMID- 14735394 TI - An unusual case of calciphylaxis. AB - BACKGROUND: Cutaneous calciphylaxis is a rare disorder that occurs most frequently in patients with end-stage renal disease (ESRD), those on hemodialysis, and renal transplant recipients. It is frequently associated with hyperparathyroidism and a markedly elevated calcium-phosphate product, and it carries a high mortality rate. The usual clinical presentation is of painful, stellate necrosis of the thighs or buttocks, often in the setting of livedo reticularis. Death usually results from septicemia. OBJECTIVE: This report documents an unusual case of recurrent, self-limiting calciphylaxis in the setting of a patient with ESRD and discusses the clinical and pathologic features of this potentially very fatal disorder. METHODS AND RESULTS: A 52-year-old woman presented with a greater than one-year history of relapsing and remitting, exquisitely painful, necrotic, numular plaques on the abdomen, breast, and arm. This patient had a markedly elevated calcium-phosphate product and parathyroid hormone level. The diagnosis of calciphylaxis was made by wedge biopsy of the most recent plaque, revealing calcification of medium-sized subcutaneous vessels and lobular capillaries with associated epidermal necrosis. CONCLUSIONS: This case demonstrates an unusual clinical variant of calciphylaxis that presented without the characteristic stellate necrosis or livedo reticularis that normally marks this condition and spontaneous resolution without incurring septicemia. Regardless of morphology, calciphylaxis should be considered in the differential diagnosis of painful, necrotic lesions occurring in the setting of ESRD. PMID- 14735395 TI - Fixed drug eruption due to influenza vaccine. AB - Two females, one 28 years old and the other 32 years old, developed dusky red macules with pain and a burning sensation within 24 hours of receiving influenza vaccine. One patient had blisters within the lesion. A clinical diagnosis of fixed drug eruption (FDE) due to influenza vaccine was made. Histopathology was consistent with the diagnosis of FDE. The lesions subsided within 2 weeks of topical corticosteroid treatment. The diagnosis was confirmed by topical provocation test with influenza vaccine. A Medline search revealed these cases to be the second report of this kind of reaction due to influenza vaccine. PMID- 14735396 TI - Chondroid syringoma with small tubular lumina. AB - BACKGROUND: Chondroid syringoma or mixed tumor of the skin is a rare epithelial tumor with eccrine differentiation. The variant with small tubular lumina is extremely uncommon. OBJECTIVE: Chondroid syringoma is usually present as a single subcutaneous nodule. This report describes a man with multiple scalp lesions exhibiting features of chondroid syringoma with small tubular lumina. METHODS: A Pathological examination of surgically resected multiple scalp nodules was carried out by routine histology and immunohistochemistry. The following antibodies were found: cytokeratin, epithelial membrane antigen, and vimentin. RESULTS: Microscopic findings revealed multiple chondroid syringoma with small tubular lumina. The tumor cells were positive for high- and low-molecular-weight keratin, epithelial membrane antigen, and vimentin. CONCLUSION: This is the first report of multiple chondroid syringoma with small tubular lumina. PMID- 14735399 TI - Axillary hidradenitis suppurativa: a further option for surgical treatment. AB - BACKGROUND: Hidradenitis suppurativa is a chronic inflammatory disease of the cutis with furuncles, fistulas, and abscesses. The disease is mostly located in groin and axilla. As conservative treatment can usually not prevent recurrence, surgical treatment is the method of choice. METHODS: We report on 20 patients with axillary hidradenitis suppurativa. The inflammatory region was excised in a rhomboid shape and immediately covered with a transposition flap according to Limberg. Postoperatively, all patients received antibiotic treatment and immobilization of the arm. Physiotherapy started after 2 weeks. RESULTS: No flap complications occurred. The functional and aesthetic results were very satisfactory. Movement of shoulder showed no restrictions. A recurrence with single fistulas was seen in 3 patients. CONCLUSIONS: Conservative treatment of hidradenitis suppurativa is followed by a high rate of recurrence. Only radical debridement offers a cure. The therapy of choice is the radical excision of the affected region and immediate coverage with a flap. Open granulation or split skin grafting often results in a prolonged hospitalization, higher morbidity, and functional problems. Thus, open granulation is inferior to primary closure by a transposition flap. Using the Limberg flap, the donor site is allowed to be closed primarily. PMID- 14735400 TI - Safety of topical minoxidil solution: a one-year, prospective, observational study. AB - BACKGROUND: Topical minoxidil solution (TMS) is widely used for androgenetic alopecia (AGA), and this is the first report of a large safety trial. OBJECTIVES: The aim of the study was to evaluate the safety profile of TMS by comparing hospitalization and death rates among subjects using TMS with controls. Cardiovascular safety and pregnancy outcomes were evaluated, and usage patterns were described. METHODS: All subjects were followed at baseline, 3, 6, 9, and 12 months. Usage patterns, pregnancy status, overnight hospital stays, and cardiovascular risk factors were evaluated. Subjects rated effectiveness of TMS in the treatment of AGA. Statistical analyses were conducted to determine if TMS was associated with an increased risk of death or hospitalization. RESULTS: TMS is a safe and effective treatment for AGA. There were no increases in cardiovascular events and no apparent increased risk for adverse pregnancy outcomes. CONCLUSIONS: This large, prospective study demonstrated the overall safety of TMS in the treatment of AGA. PMID- 14735402 TI - Quantitation of human immunodeficiency virus type 1 RNA loads in cervicovaginal secretions in pregnant women and relationship between viral loads in the genital tract and blood. AB - The purpose of this study was to analyze the quantitation of the human immunodeficiency virus type 1 RNA (HIV-1 RNA) in the genital tract of HIV-1 infected pregnant women and to evaluate a possible correlation with the viral load in blood plasma (Spearman's rank correlation coefficient). A total of 38 each of cervical, vaginal, and blood samples from 38 women were obtained during the third trimester of pregnancy for quantitation of the HIV-1 RNA load. Viral loads were determined by reverse transcription-polymerase chain reaction. The HIV 1 RNA viral load was detectable in 29 of the 38 (76.3%) blood samples, in 6 of the 38 (15.7%) cervical secretion samples, and in 8 of the 38 (21%) vaginal secretion samples. Overall, the correlation between the HIV-1 RNA viral load in the blood plasma and in cervical secretion samples was 0.51 ( P<0.001). However, the correlation disappeared ( r=0.27) when three patients with high blood plasma viral loads were eliminated from the statistical study. The viral load in the vaginal secretions did not correlate with that in the blood samples ( r=0.26). There were two cases in which HIV-1 RNA was undetectable in the blood and cervix but was detectable in vaginal secretions: one woman had 220 copies/ml and the other 68 copies/ml. These results suggest that pregnant women with undetectable viral loads in blood plasma are still at risk of transmitting the virus vertically during vaginal delivery. Because of this, antiretroviral prophylaxis during vaginal delivery must be administered to HIV-1-infected women and their newborns, regardless of the mother's viral load in plasma. In conclusion, quantification of cervicovaginal levels of HIV-1 may represent a useful tool for assessing the individual risk associated with a vaginal delivery and for guiding decisions about whether a scheduled caesarean should be recommended. PMID- 14735401 TI - Are there patients with peritonitis who require empiric therapy for enterococcus? AB - Enterococci are an increasingly important cause of nosocomial infections. While the clinical impact of enterococci in cases of bacteremia and super-infections in selected patient populations has been well-established, their role as primary pathogens in polymicrobial intra-abdominal infections remains controversial. While it has been suggested that the presence of enterococci increases the rate of infectious post-operative complication, it has also been demonstrated that polymicrobial intra-abdominal infections involving enterococci can be treated successfully with appropriate surgical drainage and antibiotics, such as cephalosporins, that are not active against enterococci. Therefore, the question arises of whether or not antibiotic coverage against enterococci should be included in the empirical treatment of peritonitis in certain high-risk patient populations. An extensive literature review revealed some evidence arguing in favour of using empirical therapy with enterococcal coverage for intra-abdominal infections in the following cases: (i) immunocompromised patients with nosocomial, post-operative peritonitis; (ii) patients with severe sepsis of abdominal origin who have previously received cephalosporins and other broad spectrum antibiotics selecting for Enterococcus spp.; (iii) patients with peritonitis and valvular heart disease or prosthetic intravascular material, which place them at high risk of endocarditis. The ideal therapeutic regimen for these high-risk patients remains to be determined, but empirical therapy directed against enterococci should be considered. PMID- 14735403 TI - Immune modulation with a staphylococcal preparation in fibromyalgia/chronic fatigue syndrome: relation between antibody levels and clinical improvement. AB - The aims of this study were to evaluate the serological response to treatment with staphylococcal vaccine in fibromyalgia/chronic fatigue syndrome patients and to explore the relationship between serological response and clinical effect. Twenty-eight patients, half of whom served as controls, were recruited from a 6 month randomised trial in which repeated administration of the staphylococcal toxoid vaccine Staphypan Berna (Berna Biotech, Switzerland) was tested against placebo. Antibody status against extracellular toxins/enzymes, cell-wall components, and enterotoxins was evaluated at baseline and at endpoint. The clinical response to treatment was recorded in rating scales. In the group receiving active treatment, significant serological changes were recorded, whereas no significant changes were found in controls. Treatment led to a significantly increased capacity of serum to neutralise alpha-toxin and a significant increase in serum IgG to alpha-toxin and lipase. Furthermore, the increase in these parameters combined paralleled the improvement in clinical outcome. Thus, the greater the serological response, the greater was the clinical effect. In conclusion, this explorative study has shown that repeated administration of the Staphypan Berna vaccine in patients with fibromyalgia/chronic fatigue syndrome causes a serological response to several staphylococcal antigens, particularly to certain extracellular toxins and enzymes. The results further show that this response is related to the clinical outcome of treatment. PMID- 14735404 TI - Subcutaneous administration of a recombinant vaccinia virus vaccine expressing multiple envelopes of HIV-1. AB - A critical goal of HIV vaccine development is the identification of safe and immunogenic vectors. Recombinant vaccinia virus is a highly effective vaccine vector, with demonstrated capacity to protect animals from various viral pathogens, including rabies. Unlike many other candidate vaccine vectors, vast human experience exists with the parenteral smallpox vaccine. However, consideration of recombinant vaccinia virus as a modern vaccine is complicated by the relatively high prevalence of immunocompromised persons compared to such prevalence 4 or more decades ago (when smallpox vaccination was still routine). Administering vaccine by the subcutaneous (SQ) route, rather than the traditional scarification route, could address these concerns. SQ administration could prevent transmission of vaccinia virus to potentially vulnerable persons; it could also avoid the most common adverse events, which are cutaneous in nature. However, previous studies suggest that elicitation of immune response against passenger gene products following SQ administration requires development of a superficial pox lesion, defeating the intention of SQ administration. This is the first report to demonstrate that SQ administration of recombinant vaccinia virus does elicit immune response to the passenger protein in the absence of a cutaneous pox lesion. Results further show that a multi-envelope HIV vaccine can elicit antibody responses toward heterologous HIV-1 not represented by primary sequence in the vaccine. These findings have global implications because they support the consideration of recombinant vaccinia virus as a valuable HIV vaccine vector system. PMID- 14735405 TI - Impact of drug-resistant Mycobacterium tuberculosis on treatment outcome of culture-positive cases of tuberculosis in the Archangel oblast, Russia, in 1999. AB - The objective of this study was to evaluate the outcome of treatment of culture positive cases of tuberculosis registered in Archangel, Russia, in 1999, and to analyse the influence of Mycobacterium tuberculosis drug resistance on treatment outcome. The outcome of tuberculosis treatment was evaluated for 235 new and 61 previously treated culture-positive cases diagnosed in 1999. Of the 235 new cases, there were 150 (63.8%) cases of treatment completion, 20 (8.5%) cases of treatment failure, 29 (12.3%) cases of death during treatment, and 29 (12.3%) cases in which the patient failed to pick up medications for at least 2 consecutive months. The outcome in 7 (3%) cases was unknown, as the patients were transferred outside the oblast region. Among the 61 previously treated cases, the rate of treatment completion was low (26.2%), and rates of treatment failure (23%) and failure to pick up medications for at least 2 consecutive months (29.5%) were high. The relation between the susceptibility pattern of the infecting strain as determined by the Bactec method and tuberculosis treatment outcome was analysed for 76 patients. The majority (69%) of patients infected with drug-susceptible strains was cured. A large proportion (58.8%) of patients infected with Mycobacterium tuberculosis resistant to more than two drugs did not respond to treatment, i.e. the treatment failed or the patients died. The high rates of death (16.7%) and failure (66.7%) among patients infected with multidrug resistant strains illustrate the negative impact of multidrug resistance on the outcome of tuberculosis treatment. Pan-resistance was significantly associated with treatment failure (P<0.001). The spread of resistant Mycobacterium tuberculosis has a serious negative impact on the outcome of tuberculosis treatment in Archangel, Russia. PMID- 14735406 TI - Recalling sexual behavior: a methodological analysis of memory recall bias via interview using the diary as the gold standard. AB - This study examined the effect of time lag on the validity of retrospective self reports of sexual behavior. Seventy-five heterosexual students (44 women, 31 men) made daily recordings of sexual behavior, condom use, and alcohol or substance use for 1 month. Ability of respondents to recall sexual behavior recorded during this period was assessed at 1, 2, and 3 months after diary completion using recall interviews (25 interviewed at each interval). For vaginal intercourse, total recall error was significantly greater at 3 months than at 1 month post diary. For all other variables assessed, the 2- and 3-month time intervals did not produce significant increase in total recall error. Higher frequency of vaginal intercourse, orgasm, and alcohol use prior to sexual activity were associated with total recall error for some but not all behaviors and outcomes. The results provide a partial validation of the diary-interview recall model as a method for studying recall error. PMID- 14735407 TI - Intercourse debut age: poor resources, problem behavior, or romantic appeal? a population-based longitudinal study. AB - The most important predictors of early intercourse debut are reported to be poor social resources and early developing problem behaviors. In this study we have a new, additional emphasis on variables related to self-concept and social acceptance. In a population-based longitudinal study, 1399 Norwegians were followed over a 7-year span. We analyzed data using multivariate Cox-regression techniques. Early intercourse debut was part of a broader spectrum of problem behaviors, including early alcohol intoxication and early-developing conduct problems. A new finding was that a positive self-concept in the domain of romantic appeal was also a strong predictor, but only for boys. We suggest that the findings may have important implications for prevention and more research should be conducted along this line. PMID- 14735408 TI - Partner influence on noncondom use: gender and ethnic differences. AB - This study investigates gender and ethnicity differences in the experience of not using a condom due to a partner s influence (unwanted noncondom use). Analysis of 247 anonymous questionnaires from students at urban community college campuses revealed that 46.7% had experienced unwanted noncondom use since age 16, and 37% had experienced unwanted noncondom use with their current or most recent partner. Males and females reported equal levels of unwanted noncondom use. However, African-American and Latino participants reported higher levels of unwanted noncondom use than Whites. The findings indicate that females, males, and people from ethnic groups at high risk for HIV infection need support to carry out their safer sex intentions. PMID- 14735409 TI - Sexuality attitudes: the impact of trauma. AB - This study examines the relationship between traumatic events and attitudes toward sexuality. Our results show that suffering trauma is related to more accepting attitudes concerning sexuality. Generally, people who suffer negative events, many of which are traumatic, are more likely to see both pornography and having a homosexual friend or family member as acceptable. Traumatic events that are sex-related or related to other physical assault proved to be most significant in the prediction of sexuality attitudes for women only. The results are specified by gender: Trauma predicts attitudes toward pornography for women but not for men, and traumatic events are associated with attitudes concerning homosexuality for women. These results are discussed in light of the previous research, and suggestions for future research made. PMID- 14735410 TI - Coital debut: the role of religiosity and sex attitudes in the Add Health Survey. AB - Recent reviews suggest that religiosity is associated with the delay of adolescent coital debut (Rostosky, Wilcox, Wright, Randall, in press; Wilcox, Rostosky, Randall, Wright, 2001). Few studies, however, have examined this association using longitudinal data to test theoretically driven models. We analyzed data from 3,691 adolescents (ages 15-21), testing the hypothesis that adolescent religiosity and sex attitudes (at Wave 1) predict later coital debut (at Wave 2) and that these predictive relationships vary by gender. Findings indicated that beyond demographic factors and number of romantic partners, religiosity reduced the likelihood of coital debut for both males and females. After accounting for the effects of religiosity, anticipation of negative emotions after coital debut (sex attitude factor 1) further reduced its likelihood for females and males. Finally, adolescent girls - but not boys - who anticipated positive emotions following sexual intercourse (sex attitude factor 2) were more likely to debut. While virginity pledge status was associated with coital debut for boys and girls, more conservative beliefs about sex appeared to mediate its effect. Finally, a significant interaction between race and religiosity indicated that African American adolescent males who had either signed a virginity pledge or were more religious were significantly more likely to debut than both White non-Hispanic males and African American males who were less religious and/or who had not signed a pledge. The implications of these results for adolescent pregnancy prevention programming are discussed. PMID- 14735411 TI - Sexual modes questionnaire: measure to assess the interaction among cognitions, emotions, and sexual response. AB - The goal of the present article is to present a new measure developed to assess cognitive and emotional factors of sexual function. This instrument was developed especially to test some hypotheses derived from Beck s new theoretical conceptualization (the modes theory; A. T. Beck, 1996). This model, characterized by its systemic and integrated approach, constitutes a remarkable development from a linear to a network perspective of the cognitive-emotional-behavioral processes. The new concept of mode, as a composite of schemas (cognitive, emotional, and behavioral) interacting together, is theoretically sound and supported by recent research findings from clinical and experimental sets (see A. T. Beck, 1996, for a revision). Our aim is to develop a new measure specifically created to assess these integrated and interdependent processes in the field of sexuality. The Sexual Modes Questionnaire (SMQ; male and female versions) is a combined measure constituted by three interdependent subscales: automatic thoughts, emotions, and sexual response presented during sexual activity. Psychometric studies showed good reliability and validity results in both versions, and high correlations between several dimensions of the three subscales support the concept of mode and its interactional character. Moreover, the capacity showed by the SMQ to discriminate between sexually functional and dysfunctional subjects and its high correlations with measures of sexual functioning emphasize the role of cognitive-emotional processes on sexual problems, supporting the clinical value of the measure. PMID- 14735412 TI - Toward improved interpretation and theory building of African American male sexualities. AB - This paper examined five challenges to clear understanding of African American male sexualities: incorrect assumptions of African American homogeneity; an underemphasis on developmental change, the contexts and the meanings of sexual behaviors; and a lack of compelling theoretical grounding for African American sexualities. Critical elements for effective theorizing and research about African American sexualities (i.e. multiple levels of analysis, examination of phenomenological meaning of sexuality, measurement of dynamic/developmental change) were outlined and candidate theories within sexual science (social exchange theories, symbolic interactionism, sexual scripting theory) were analyzed in light of these elements. It is suggested that a re-orientation of sex research about African American men using these elements will result in improved understanding of African American sexualities in multiple contexts. PMID- 14735413 TI - Characteristics of men and women who complete or exit from an on-line internet sexuality questionnaire: a study of instrument dropout biases. AB - This study compared respondents who completed an Internet sexuality questionnaire and those who dropped out before completion. The study was in Swedish and comprised 3,614 respondents over a 2-week period (53% males, 47% females). There were significant differences between males, of whom 51% dropped out before completion of the 175-item questionnaire, and females, of whom 43% dropped out. Dropout in both genders followed a curve of negative acceleration. The data suggest that dropout is likely to be significant and gender and demographically biased, and to occur significantly earlier for men than for women. Geography, education, sexual orientation, age, relationship status, living arrangements, and Internet connection speed were related to dropout for men, while only relationship status and living arrangements, which were in the opposite direction from men, were related to dropout in women. PMID- 14735414 TI - [The influence of residual tumor on local recurrence after unplanned resection of soft tissue sarcoma]. AB - BACKGROUND AND OBJECTIVE: The "unplanned" excision of a soft tissue sarcoma (STS) is defined as its resection without preoperative staging procedures and without aiming at wide resection margins owing to lacking knowledge of the malignancy of the tumor. Aim of this study was to evaluate the influence of such unplanned excisions, in spite of immediate re-excision, upon local recurrence. PATIENTS AND METHODS: Results in 110 consecutive patients were analysed. In all of them an unplanned resection of a STS of the limbs or trunk not achieving wide resection margins had been performed prior to referral. End-point of statistical analysis was local relapse. Median follow-up time was 82 months. RESULTS: Residual tumor was found in 34 specimens of primary re-excisions (31 %). Local recurrence occurred in 14 patients, 23 patients developed distant metastases. Multivariate analysis revealed two significant prognostic factors for local recurrence: residual tumor in the re-excised specimen and occurrence of distant metastases. CONCLUSIONS: Presence of residual tumor in re-excised specimens is a prognostic factor for local tumor relapse, is the development of distant metastases. Suspicious soft tissue tumors should be excised after oncological regimens, preferably in experienced centers. PMID- 14735415 TI - [Diagnosis of premenstrual disorders]. AB - BACKGROUND AND OBJECTIVE: There is no valid method in the German literature for assessing premenstrual disorders. This study was undertaken to translate into German, validate and test the reliability of the "Premenstrual Syndrome Tension Observer/Self-Rating Scale" and the "Visual Analogue Scale for Premenstrual Disorders". PATIENTS AND METHODS: 55 patients diagnosed with premenstrual dysphoric disorder took part in the study. After fullfilling the inclusion criteria, sociodemographic and medical data were studied. The self-assessment by PMTS-SR and VAS-PmD was done two days premenstrual and nine days postmenstrual by all 55 patients. The objective assessment (done by a clinical interview and the PMTS-O as the reference standard) was obtained afterwards without knowing the self-assessment. RESULTS: The average age of the tested women was 35 years. The average increase of 85 % in premenstrual symptoms (PMTS-O and PMTS-SR scale) indicates a group with severe premenstrual disorders. The correlation coefficient between PMTS-O and PMTS-SR scale, pre- and postmenstrually, was 0.83 and 0.84, respectively (p < 0.0001 for both). The correlation coefficient between PMTS-O and VAS-PmD, pre- and postmenstrually, was 0.5 and 0.69, respectively (p < 0.0001 for both). This was calculated at 0.64 and 0.67, respectively (p < 0.0001 for both) between PMTS-SR and VAS-PmD. Reliability analysis for PMTS-O was, pre- and postmenstrually, 0.71 and 0.29, respectively, for PMTS-SR 0.77 and 0.78, respectively, and for VAS-PmD it was 0.79 and 0.87, respectively. CONCLUSIONS: The German version of the PMTS-O and the PMTS-SR scale are "application friendly" as well as proven to be a valid and reliable method for supporting the diagnosis, course of disease and research aspects concerning premenstrual disorders. Initially VAS-PmD should be used in combination with PMTS-O and PMTS-SR scale. PMID- 14735416 TI - [Lambert-Eaton myasthenic syndrome with acute respiratory failure associated with small cell lung cancer]. AB - HISTORY AND ADMISSION FINDINGS: A 53-year-old woman had suffered from progressive lower limb weakness for 8 weeks. She was not able to walk on her own. Two hours after admission endotracheal intubation and mechanical ventilation became necessary because of acute respiratory failure. INVESTIGATIONS: Chest radiograph and computerized tomography showed a huge lesion originating from the lower lobe of the left lung with mediastinal infiltration. Histological examination of bronchoscopical-guided biopsy disclosed small cell lung cancer. Electrophysiological studies revealed an incremental response on high-rate stimulation in the repetitive nerve stimulation test. Autoantibodies to P/C-type voltage-gated calcium channels were detected. Further, laboratory test results revealed hyponatremia (100 mmol/l) and elevated ADH (11 pg/ml). DIAGNOSIS, TREATMENT AND CLINICAL COURSE: These results led to the diagnosis of paraneoplastic Lambert-Eaton myasthenic syndrome (LEMS) with acute respiratory failure caused by small cell lung cancer. Additionally, SIADH (syndrome of inappropriate antidiuretic hormone secretion) was diagnosed as another paraneoplastic disease. Because of a prolonged weaning process resulting from inadequacy of mechanical ventilation, administration of intravenous immunoglobulins and chemotherapy with cisplatin and etoposide were undertaken. This made successful extubation possible after 7 days. In spite of all supportive treatment, clinical deterioration occurred. The patient died 27 days after the small cell lung cancer had been diagnosed. CONCLUSION: The Lambert-Eaton myasthenic syndrome should be considered in cases of unexplained acute respiratory failure and clinical evidence of neoplastic disease. PMID- 14735417 TI - [Dementias: communication between doctor, patient and relatives]. PMID- 14735418 TI - [Emerging risk factors for cardiovascular disease]. PMID- 14735420 TI - [Changes in risk factors after inpatient rehabilitation of patients with coronary heart disease (KAROLA)]. PMID- 14735419 TI - [Fixed combination of inhalant steroids and long-acting beta(2) agonists]. PMID- 14735421 TI - [Chronic lead poisoning caused by Ayurvedic drugs]. PMID- 14735424 TI - Dynamic and functional imaging of the musculoskeletal system. AB - Conventional magnetic resonance imaging (MRI) examination of the musculoskeletal system is limited to static evaluation of muscle, spine, and joint morphology. However, using the variety of contrast mechanisms available with MRI, it is possible to obtain functional and biomechanical data about the musculoskeletal system. The function of joints and muscles are for motion and weight bearing, so pathology that only occurs during activity can be missed on static MRI examination. Similarly, important information about muscle biomechanics, muscle energetics, and joint function can be obtained with unique MRI contrast such as T(2)-mapping, spectroscopy, blood-oxygenation-level-dependent (BOLD) imaging, and molecular imaging. These new techniques hold the promise for a more complete and useful joint examination. PMID- 14735425 TI - Functional assessment of the joints using kinematic magnetic resonance imaging. AB - Kinematic magnetic resonance imaging (MRI) techniques were developed in recognition of the fact that certain pathologic conditions that affect the joints are position dependent or associated with stress or "loaded" conditions. Information obtained using kinematic MRI procedures often serves to definitively identify and characterize the underlying abnormality or to supplement the findings acquired with standard MRI techniques. To date, kinematic MRI methods have been applied to evaluate virtually every articulation. This article presents information for kinematic MRI applications applied to the temporomandibular and patellofemoral joints, with a discussion of the normal kinematics and pathokinematics seen using these unique imaging procedures. Other kinematic MRI procedures applied to the ankle, wrist, cervical spine, shoulder, and lumbar spine are briefly discussed. PMID- 14735426 TI - Magnetic resonance imaging of the weight-bearing spine. AB - Weight-bearing magnetic resonance (MR) imaging of the spine can either be simulated by imaging the patient in the supine position in combination with a special axial loading device or be achieved by using vertically open configuration MR systems, which allow for in vivo MR images of the spine under upright weight-bearing conditions in either seated or standing body positions. Weight-bearing MRI of the spine permits the study of physiological as well as pathological changes in the relationships of the intervertebral disk, the spinal canal, and the neural foramina as well as the assessment of segmental instability in physiologic body positions. With this technique, MR images may be taken in painful body positions so that morphological changes of the intervertebral disk or other spinal structures may be correlated with pain or other symptoms. In selected cases, weight-bearing MRI of the spine may demonstrate clinically relevant neural compromise or foraminal stenosis, which may be occult on conventional MR images obtained in the supine position. PMID- 14735427 TI - Cine phase-contrast magnetic resonance imaging as a tool for quantification of skeletal muscle motion. AB - In recent years, biomechanics researchers have increasingly used dynamic magnetic resonance imaging techniques, such as cine phase contrast (cine PC), to study muscle and bone motion in vivo. Magnetic resonance imaging provides a non invasive tool to visualize the anatomy and measure musculoskeletal tissue velocities during joint motion. Current application of cine PC magnetic resonance imaging in biomechanics includes study of knee joint kinematics, tendon strain, and skeletal muscle displacement and shortening. This paper article reviews the use of cine PC magnetic resonance imaging for quantification of skeletal muscle motion. The imaging studies presented examine the relative motion of the knee flexor and extensor muscles after orthopedic surgery and examine the uniformity of shortening within the biceps brachii muscle. The current challenges and limitations of using cine PC magnetic resonance imaging in biomechanics research are addressed as well as opportunities for future studies of skeletal muscle motion using dynamic magnetic resonance imaging. PMID- 14735428 TI - T2 mapping of muscle. AB - Muscle activation produces increases in magnetic resonance ( T(2)) signal intensity leading to recruitment images that demonstrate spatial patterns and intensity of muscle activation. These T(2) activation maps are useful for visualizing and quantifying various aspects of muscle function. Activity dependent changes in T(2) result from an increase in the T(2) relaxation time of muscle water. The current state of investigation indicates that the mechanism of increased T(2) results from osmotically driven shifts of muscle water that increase the volume of the intracellular space and from intracellular acidification resulting from the end products of metabolism. Although the spatial resolution of magnetic resonance imaging is still insufficient to map territories of individual motor units, it is possible to demonstrate nonuniform activation between subregions or compartments of muscle. Taken together, the attributes of the T(2) mapping technique hold great potential for demonstrating aberrant muscle activation patterns in pathology and positive adaptation to exercise or rehabilitative intervention. PMID- 14735429 TI - BOLD magnetic resonance imaging of skeletal muscle. AB - Blood-oxygen-level-dependent (BOLD) imaging was a concept introduced in 1990 for evaluating brain activation. The method relies on magnetic resonance imaging (MRI) contrast resulting from changes in the microvascular ratio of oxyhaemoglobin (oxyHb) to deoxyhaemoglobin (deoxyHb). OxyHb is diamagnetic, whereas deoxyHb is paramagnetic, which produces a local bulk magnetic susceptibility effect and subsequent MRI signal change. The changes are typically observed in T(2)*-weighted functional MRI scans. However, there has recently been interest in BOLD as a way to evaluate microcirculation of any normal or diseased tissue. This review focuses on the application of BOLD imaging in the understanding of normal and diseased skeletal muscle. In addition we present new findings showing the possible application of BOLD imaging with hyperoxia for evaluating skeletal muscle physiology. PMID- 14735430 TI - Molecular imaging of musculoskeletal diseases. AB - Chronic musculoskeletal diseases such as arthritis, malignancy, chronic injury/ inflammation, and chronic musculoskeletal pain often pose challenges for current clinical imaging modalities. There is hope that a growing field, referred to as "molecular imaging," will shed new light on these chronic phenomenon as it aims to noninvasively detect special molecular and physiologic effects such as metabolism rate, specific proteins, cell death, and particular gene-related events. Molecular imaging represents recent advances in imaging technology, engineering, chemistry, molecular biology, and genetics that have coalesced into a multidisciplinary and multimodality effort. Molecular probes are currently being developed not only in radionuclide-based techniques but also in magnetic resonance imaging, magnetic resonance spectroscopy, ultrasound, and the emerging field of optical imaging. Furthermore, molecular imagers are fueling the development of novel molecular therapies and gene therapy, as tracking these efforts in living subjects is now possible with molecular imaging protocols. PMID- 14735431 TI - Relevant hepatotoxic effects of kava still need to be proven. A statement of the Society for Medicinal Plant Research. PMID- 14735432 TI - Anti-inflammatory compounds of plant origin. Part I. Action on arachidonic acid pathway, nitric oxide and nuclear factor kappa B (NF-kappaB). AB - Over the last 10 years, a significant body of evidence has emerged indicating that chemically diverse classes of naturally-occurring substances derived from higher plants are of potential interest for therapeutic interventions in several inflammatory diseases. Part I of this review article focuses on our current knowledge of the mechanisms by which a large range of plant-derived constituents interfere with three relevant targets involved in the inflammatory process, namely arachidonic acid metabolite pathways, nitric oxide and NF-kappaB, and discusses their potential therapeutic use in the management of relevant inflammatory diseases. PMID- 14735433 TI - Theasinensin A, a tea polyphenol formed from (-)-epigallocatechin gallate, suppresses antibiotic resistance of methicillin-resistant Staphylococcus aureus. AB - When (-)-epigallocatechin gallate (EGCG), the main constituent of tea polyphenols, was kept in a neutral buffer, it decomposed rapidly to give theasinensin A as the major product. Theasinensin A suppressed the oxacillin resistance of methicillin-resistant Staphylococcus aureus (MRSA). In the presence of theasinensin A (3.5 x 10(-5) M), the minimum inhibitory concentrations (MICs) of oxacillin decreased from 256 or 64 microg/mL to 4 microg/mL for the MRSA strains used. The presence of this compound (3.5 x 10(-5) M) also decreased the MIC of other beta-lactam (penicillin G, from 32 microg/mL to 0.125-0.5 microg/mL; ampicillin, from 16-32 microg/mL to 0.5-1 microg/mL) and aminoglycoside (streptomycin, from 4 - 16 microg/mL to 0.125-4 microg/mL) antibiotics for the MRSA strains. PMID- 14735434 TI - The chalcone butein from Rhus verniciflua shows antifibrogenic activity. AB - Butein is known to be the major component of the bark of Rhus verniciflua Stokes (Anacardiaceae). The aim of this work was to investigate the effects of butein on liver fibrosis induced by carbon tetrachloride (CCl4) in rats, and to explore its antifibrogenic mechanism. Butein (10 mg/kg/day or 25 mg/kg/day) showed a significant reduction of hydroxyproline and malondialdehyde levels in rats. The expression of alpha1(I) collagen and tissue inhibitor of metalloproteinase-1 (TIMP-1) mRNAs in liver was clearly reduced in a dose-dependent manner in rats given butein compared with control CCl4-treated rats. These data suggest the potential of butein to serve as an antifibrogenic agent by inhibition of collagen accumulation and lipid peroxidation, and by down-regulation of the expression of both alpha1(I) collagen and TIMP-1 mRNA. PMID- 14735435 TI - Effects of the flavonoid quercetin and its methylated metabolite isorhamnetin in isolated arteries from spontaneously hypertensive rats. AB - Chronic oral quercetin exerts antihypertensive effects in spontaneously hypertensive rats (SHR). In the present study, the vasodilator effects of the flavonoid quercetin and its main metabolite isorhamnetin were analysed in isolated thoracic aorta, iliac artery and on the isolated perfused mesenteric resistance vascular bed from SHR and normotensive Wistar Kyoto rats (WKY). In noradrenaline-precontracted vessels from SHR there was an inverse correlation between the relaxant potency (pIC50) of quercetin (4.76 +/- 0.02, 5.08 +/- 0.12, 5.30 +/- 0.18, in aorta, iliac arteries and mesentery, respectively) and isorhamnetin (4.90 +/- 0.11, 5.38 +/- 0.15 and 5.80 +/- 0.10, respectively) and the diameter of the vessel studied. Both flavonoids were more potent in endothelium-denuded aortae and iliac arteries from SHR than from normotensive WKY rats. In addition, in aortae from SHR both flavonoids restored the endothelial dependent vasodilation. Isorhamnetin, but not quercetin, also reduced the endothelium-dependent contractile responses induced by acetylcholine. These direct vasodilator effects, together with the improvement of endothelial function, are good candidates to explain the blood pressure reduction and vascular protective effects of quercetin in animal models of hypertension and possibly in human cardiovascular diseases. PMID- 14735436 TI - Antinociceptive effects of ginsenosides injected intracerebroventricularly or intrathecally in substance P-induced pain model. AB - We have examined the effects of several ginsenosides (Rb1, Rb2, Rc, Rd, Re, Rf, Rg1 and Rg3) administered intracerebroventricularly (i.c.v.) or intrathecally (i.t.) on the nociceptive behavior induced by substance P (0.7 microg) injected i.t. Among the several ginsenosides studied, Rb2, Rc, Rd, and Re, but not Rb1, Rf, Rg1 and Rg3, treated i.c.v. (50 microg) attenuated the nociceptive behavior induced by substance P injected i.t. On the other hand, we found that i.t. treatment with 50 microg of Rb1, Rb2, Rd, or Rf effectively attenuated the nociceptive behavior induced by i.t. injected substance P. However, the i.t. treatment with the same doses of Rc, Re, Rg1 or Rg3 was not effective for antagonizing i.t. injected substance P-induced nociceptive behavior. Our results show that ginsenosides Rb2, Rc, Rd, or R2 injected supraspinally exert a antinociceptive effect in the substance P-induced pain model. Furthermore, Rb1, Rb2, Rd, or Rf treated spinally produce antinociception in the substance P induced pain model. PMID- 14735437 TI - Transgenic ginseng cell lines that produce high levels of a human lactoferrin. AB - In order to produce a human lactoferrin (hLf) protein in cultured plant cells, we developed Korean ginseng (Panax ginseng) cell line using an oxidative stress inducible peroxidase (SWPA2) promoter and characterized the production of human lactoferrin in cultured cells. A construct containing a targeting signal peptide from tobacco endoplasmic reticulum fused to human lactoferrin cDNA under the control of SWPA2 promoter was engineered. Transgenic Korean ginseng cell lines that produced a recombinant hLf protein were successfully generated and confirmed by PCR and Southern blot analyses. Western blot and ELISA analyses showed that hLf protein was synthesized in the transgenic cells. The production of hLf showed a maximal level (up to 3.0% of total soluble protein) in the stationary phase of callus cultures. These results suggest that the transgenic cell lines in this study will be biotechnologically useful for the commercial production of hLf protein in cell cultures, with no need for purification. PMID- 14735438 TI - Transport of parthenolide across human intestinal cells (Caco-2). AB - This study examined the intestinal epithelial membrane transport of the sesquiterpene lactone parthenolide, a bioactive compound present in the migraine prophylactic herb feverfew. The Caco-2 human colonic cell line was used as an in vitro model of the human intestinal mucosal barrier. The bidirectional transport (apical to basolateral and basolateral to apical) of parthenolide was investigated using Caco-2 monolayers grown on Transwell inserts. Quantitation of parthenolide was performed using high performance liquid chromatography (HPLC). Apical to basolateral and basolateral to apical permeability coefficients and percent transport were calculated and a potential bioavailability of parthenolide was determined. Sodium fluorescein was used as a marker for paracellular leakage. Parthenolide, at a concentration of 250 microM, demonstrated substantial linear transport across the monolayer. The transport parameters were not affected by the presence of MK-571, an inhibitor of multidrug resistance transporter P glycoprotein (MRP). Upon comparison of the transport parameters of parthenolide with atenolol under identical conditions and the reported values for model compounds like mannitol and propranolol, it is concluded that parthenolide is effectively absorbed through the intestinal mucosa via a passive diffusion mechanism. PMID- 14735439 TI - In vitro studies indicate that miquelianin (quercetin 3-O-beta-D glucuronopyranoside) is able to reach the CNS from the small intestine. AB - Miquelianin (quercetin 3-O-beta-D-glucuronopyranoside) is one of the flavonoids of St. John's wort (Hypericum perforatum L.) whose antidepressant activity has been shown by the forced swimming test, an in vivo pharmacological model with rats. However, nothing is known about its ability to reach the CNS after oral administration. We examined the pathway of miquelianin from the small intestine to the central nervous system using three in vitro membrane barrier cell systems. In the Caco-2 cell line, miquelianin showed a higher uptake (1.93 +/- 0.9 pmol x min(-1) x cm(-2)) than hyperoside (quercetin 3-O-beta-D-galactopyranoside; 0.55 +/- 0.18 pmol x min(-1) x cm(-2)) and quercitrin (quercetin 3-O-alpha-L rhamnopyranoside; 0.22 +/- 0.08 pmol x min(-1) x cm(-2)). The permeability coefficient of miquelianin (Pc = 0.4 +/- 0.19 x 10(-6) cm/sec) was in the range of orally available drugs assuming sufficient absorption from the small intestine. Uptake and permeability of the examined compounds was increased by the MRP-2 inhibitor MK-571 indicating a backwards transport by this membrane protein. Porcine cell cultures of brain capillary endothelial cells were used as a model of the blood-brain barrier (bbb) and epithelial cells of the plexus chorioidei as a model of the blood-CSF barrier (bcb). Results indicate no active transport in one direction. Although moderate, the permeability coefficients (bbb: Pc = 1.34 +/- 0.05 x 10(-6) cm/sec; bcb: Pc = 2.0 +/- 0.33 x 10(-6) cm/sec) indicate the ability of miquelianin to cross both barriers to finally reach the CNS. PMID- 14735440 TI - Transformation of ipecac (Cephaelis ipecacuanha) with Agrobacterium rhizogenes. AB - Transformed root cultures of ipecac (Cephaelis ipecacuanha A. Richard), one of the recalcitrant woody plant species for Agrobacterium-mediated transformation, were established by co-culturing of in vitro petiole segments with Agrobacterium rhizogenes ATCC 15 834. Southern blot analysis of the established roots revealed that only the TL-DNA was integrated into the plant genome without incorporation of the TR-DNA. The transformed roots grew slowly on phytohormone-free solid medium and adventitious shoots were regenerated after over 6 months of culture on HF, half-strength Murashige and Skoog (1/2 MS) medium in the dark. The individually separated transformed shoots developed into plantlets on phytohormone-free solid medium at 25 degrees C under 16 h/day light, and the plants demonstrated wider leaves, shorter internodes and vigorous root growth compared to non-transformed plants. Effects of basal media and auxins on the growth and the ipecac alkaloid production of the transformed roots were investigated either under light or in the dark. The roots cultured in the dark grew well in Gamborg B5 (B5) liquid medium containing 0.5 mg/L IBA and yielded 112 mg/L of cephaeline and 14 mg/L emetine after 8 weeks of culture. PMID- 14735441 TI - Effect of acute drought stress and time of harvest on phytochemistry and dry weight of St. John's wort leaves and flowers. AB - The phytochemistry and dry weight of cultivated St. John's wort are significantly influenced by acute drought stress and time of harvest. In this study, plants subjected to brief drought stress during both flower and seed development periods exhibited increased concentrations in 8 of the 10 phytochemicals examined in this study, including hypericin, pseudohypericin, chlorogenic acid, rutin, hyperoside, isoquercitrin, quercitrin, and quercetin. Increases ranged from 5% to 36% (hyperoside and rutin, respectively). Conversely, the concentrations of hyperforin and adhyperforin in flowers were decreased by an average of 10% in drought-stressed plants as compared to well-watered control plants. Acute drought stress decreased flower dry weight significantly during both drydown periods, although vegetative parameters (height, leaf dry weight and stem dry weight) were not adversely affected. While acute drought stress significantly altered the chemical yield in the leaves and flowers (phytochemical content x harvested dry weight), the time of harvest was the predominant factor determining phytochemical concentration in the organs of H. perforatum. PMID- 14735442 TI - Cytotoxic diterpenoids from Isodon enanderianus. AB - Five new ent-kaurane diterpenoids, enanderianins K-O (1-5), and one new ent abietane diterpenoid, enanderianin P (6), together with five known ent-kaurane diterpenoids, rabdocoetsin A (7), rabdocoetsin B (8), rabdocoetsin D (9), megathyrin A (10), megathyrin B (11) were isolated from the aerial parts of Isodon enanderianus. Their structures were determined by spectral means. Some diterpenoids were tested for their cytotoxicity against the human tumor K562 cells. Compounds 1, 2, 6, 8, 9 showed significant inhibitory activities against K562 cells with IC50 values ranging from 0.13 to 0.87 microg/mL. PMID- 14735443 TI - Three new triterpenoid saponins from Ixeris sonchifolia and their cytotoxic activity. AB - Bioassay-guided fractionation of an active n-BuOH extract of the whole plant of Ixeris sonchifolia, using a cytotoxicity assay, resulted in the isolation of three new triterpenoid saponins, ixeris saponins A (1), B (2), and C (3). On the basis of chemical evidence and extensive spectral studies, their structures were established as echinocystic acid 3-O-beta-D-glucopyranosyl(1-->3)-beta-D glucopyranosyl(1-->3)-alpha- L-arabinopyranoside (1), 3-O-[bis[beta-D glucopyranosyl(1-->2 and 1-->3)-alpha-L-arabinopyranosyl]]echinocystic acid 28-O beta-D-glucopyranosyl ester (2), and 3- O-[beta-D-glucopyranosyl(1-->3)-beta- D glucopyranosyl(1-->3)-alpha-L-arabinopyranosyl]-16alpha, 23-dihydroxyolean-12-ene 28-O-beta-D-glucopyranosyl ester (3). Compounds 2 and 3 showed significant cytotoxicity against cancer cell lines A375, L929, and HeLa with IC50 values ranging from 8.83 microM to 15.83 microM, while compound 1 was inactive against these three cell lines. PMID- 14735444 TI - Two new anti-tumor promoting serratane-type triterpenoids from the stem bark of Picea jezoensis var. jezoensis. AB - Two new serratane-type triterpenoids, 1 and 2, were isolated from the stem bark of Picea jezoensis Carr. var. jezoensis (Pinaceae). Their structures were determined to be 3beta-methoxyserrat-13-en-21beta-ol (1) and 13beta, l4beta-epoxy 3beta-methoxyserratan-21beta-ol (2) on the basis of spectroscopic methods and partial syntheses. Compounds 1 and 2 and their acetates were screened as potential anti-tumor promoters by using the in vitro short-term 12-O tetradecanoylphorbol 13-acetate (TPA)-induced Epstein-Barr virus early antigen (EBV-EA) activation assay. IC50 value evaluation showed that compound 1 was more effective than others. In addition, compounds 1 and 2 were examined for anti tumor promoting activities in a two-stage carcinogenesis assay of mouse skin tumors induced by 7,12-dimethylbenz[a]anthracene (DMBA) as an initiator and TPA as a promoter. Compounds 1 and 2 exhibited significant anti-tumor promoting effects on mouse skin carcinogenesis. PMID- 14735445 TI - Cytotoxic coumarins from Mammea harmandii. AB - Two new naturally occurring coumarins, isomesuol (1) and mammearin A (2), together with nine known Mammea coumarins 3-11 were isolated from the EtOAc extract of the leaves and twigs of Mammea harmandii. Coumarins 1, 3 and 4 showed cytotoxicity against a panel of mammalian cancer cell lines. Their structures were determined by spectroscopic methods. The assignments of 13C-NMR signals of isomesuol (1), which was isolated for the first time as a natural product, have been revised. PMID- 14735446 TI - Cytotoxic triterpenoids from the fruits of Zizyphus jujuba. AB - The following eleven triterpenoic acids were isolated from the fruits of Zizyphus jujuba (Rhamnaceae): colubrinic acid, alphitolic acid, 3-O-cis-p coumaroylalphitolic acid (3), 3-O-trans-p-coumaroylalphitolic acid (4), 3-O-cis-p coumaroylmaslinic acid, 3-O-trans-p-coumaroylmaslinic acid, betulinic acid (7), oleanolic acid, betulonic acid (9), oleanonic acid and zizyberenalic acid. The in vitro cytotoxicities of the triterpenoic acids against K562, B16(F-10), SK-MEL-2, PC-3, LOX-IMVI, and A549 tumor cell lines were investigated by the sulforhodamin B (SRB) method. Among these compounds, the lupane-type triterpenes, such as compounds 3, 4, 7, and 9, showed high cytotoxic activities. In particular, the cytotoxic activities of 3-O-p-coumaroylalphitolic acids (compounds 3 and 4) were better than those of non-coumaroic triterpenenoids (compounds 7 and 9). These results suggest that the coumaroyl moiety at the C-3 position of the lupane-type triterpene may play an important role in enhancing cytotoxic activity. PMID- 14735447 TI - Cytotoxic alkaloids from the flowers of Senna spectabilis. AB - Three new alkaloids, 3(R)-benzoyloxy-2(R)-methyl-6(R)-(11'-oxododecyl)-piperidine (3), 5-hydroxy-2-methyl-6-(11'-oxododecyl)-pyridine (4) and 5-hydroxy-2-methyl-6 (11'-oxododecyl)-pyridine N-oxide (5), together with a known alkaloid, (-) cassine (1), were isolated from the flowers of Senna spectabilis. A derivative, N,O-diacetylcassine (2), was semisynthesized. Their structures and stereochemistry were established on the basis of spectroscopic analysis. Cytotoxic activity and brine shrimp lethality of these compounds were evaluated. Compounds 2, 3 and 5 exhibited cytoxicity against KB cell lines with IC50 values of 5.2, 3.7 and 2.0 microg/mL, respectively. PMID- 14735448 TI - Tanshinone IIA from Salvia miltiorrhiza inhibits inducible nitric oxide synthase expression and production of TNF-alpha, IL-1beta and IL-6 in activated RAW 264.7 cells. AB - The inhibitory effects of tanshinone IIA, a diterpene isolated from Salvia miltiorrhiza root, on the production of nitric oxide (NO), interleukin-1beta (IL 1beta), interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha), and the expression of inducible nitric oxide synthase (iNOS) were investigated in activated RAW 264.7 cells. This compound markedly inhibited the production of NO, IL-1beta and TNF-alpha, and suppressed the expression of iNOS in a dose-dependent manner. These results suggest that the traditional use of S. miltiorrhiza as an anti-inflammatory herbal medicine may be explained, in part, by the inhibition of NO, IL-1beta, IL-6 and TNF-alpha production, and expression of iNOS. PMID- 14735449 TI - 2-(4'-aminobenzenamine)-pyrimidine, a new alpha-antagonist from Autonoe madeirensis. AB - A new natural pyrimidine derivative, 2-(4'-aminobenzenamine)-pyrimidine (1), was isolated from the bulbs of Autonoe madeirensis, a Hyacinthaceae of the Madeira Archipelago (Portugal) and the structure determined on the basis of spectroscopic data. As some pyrimidine compounds are important alpha1-adrenergic antagonists, we investigated the effect of 1 on rat vas deferens contractility induced by the alpha1-agonist phenylephrine. In concentrations virtually devoid of effect on the rat vas deferens contractility, 1 shifted to the right the concentration-response curves of phenylephrine without changing the maximal effect. These results provide evidence that 1 acts as an alpha1-adrenoceptor antagonist and suggest a competitive mechanism of action. PMID- 14735450 TI - Antioxidant activity of curtisians I-L from the inedible mushroom Paxillus curtisii. AB - Four p-terphenyl derivatives named curtisians I-L (1-4) were isolated from the MeOH extract of the fruit bodies of Paxillus curtisii. Their structures were elucidated by 2D NMR, MS, IR and UV spectroscopy and chemical reaction. The absolute configurations of curtisians were determined by GC-MS on a chiral column of the hydrolyzed products with authentic samples. Their antioxidative activities were also estimated by examination of the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical-scavenging effect. PMID- 14735451 TI - A new indolizinone from Polygonatum kingianum. AB - A new indolizinone, namely kinganone (1) together with 3-ethoxymethyl-5,6,7,8 tetrahydro-8-indolizinone (2) and isomucronulatol (3) were isolated from the rhizome of Polygonatum kingianum. Their structures were elucidated mainly on the basis of spectral data. Indolizinones 1 and 2 showed weak antibacterial and antifungal activities when compared to rifampicin and amphotericin, respectively, in the agar diffusion assay. PMID- 14735452 TI - Isolation of mangiferin from Bombax malabaricum and structure revision of shamimin. AB - Repeated column chromatography of the n-BuOH fraction of the 70% EtOH of the dried leaves of Bombax malabaricum led to the isolation of mangiferin, a xanthone. Mangiferin was identified by UV, 1H- and 13C-NMR spectroscopy and electrospray mass spectrometry. It was found to be identical to shamimin, a compound for which originally a flavonol structure was proposed, and the structure of which has to be revised. PMID- 14735455 TI - Impact of functional genomics and proteomics on radionuclide imaging. AB - The assessment of gene function following the completion of human genome sequencing may be performed using radionuclide imaging procedures. These procedures are needed for the evaluation of genetically manipulated animals or newly designed biomolecules, which requires a thorough understanding of physiology, biochemistry, and pharmacology. The experimental approaches will involve many new technologies, including in vivo imaging with single photon emission computed tomography and positron emission tomography. 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. Visualization of in vivo reporter gene expression can be performed using radiolabeled substrates, antibodies, or ligands. Combinations of specific promoters and in vivo reporter genes may deliver information about the regulation of the corresponding genes. Furthermore, protein-protein interactions and activation of signal transduction pathways may be visualized noninvasively. The role of radiolabeled antisense molecules for the analysis of messenger ribonucleic acid (RNA) content has to be investigated. However, possible applications are therapeutic intervention using triplex oligonucleotides with therapeutic isotopes, which can be brought near to specific deoxyribonucleic acid sequences to induce deoxyribonucleic acid strand breaks at selected loci. Imaging of labeled siRNA makes sense if these are used for therapeutic purposes to assess the delivery of these new drugs to their target tissue. Pharmacogenomics will identify new surrogate markers for therapy monitoring, which may represent potential new tracers for imaging. Drug distribution studies for new therapeutic biomolecules are needed at least during preclinical stages of drug development. New treatment modalities, such as gene therapy with suicide genes, will need procedures for therapy planning and monitoring. Finally, new biomolecules will be developed by bioengineering methods, which may be used for the isotope-based diagnosis and treatment of disease. PMID- 14735456 TI - The Na/I symporter (NIS): imaging and therapeutic applications. AB - The Na(+)/I(-) symporter (NIS) is the plasma membrane glycoprotein that mediates the active uptake of I(-) in the thyroid, ie, the crucial first step in thyroid hormone biosynthesis. NIS also mediates I(-) uptake in other tissues, such as salivary glands, gastric mucosa, and lactating (but not nonlactating) mammary gland. The ability of thyroid cancer cells to actively transport I(-) via NIS provides a unique and effective delivery system to detect and target these cells for destruction with therapeutic doses of radioiodide. Breast cancer is the only malignancy other than thyroid cancer to have been shown to functionally express NIS endogenously. The considerable potential diagnostic and therapeutic use of radioiodide in breast cancer is currently being assessed. On the other hand, exogenous NIS gene transfer has successfully been carried out into a variety of other cell lines and tumors, including A375 human melanoma tumors, and SiHa cervix cancer, human glioma, and hepatoma cell lines. Most notably, significant radioiodine therapy results have been obtained in the NIS-transfected human prostatic adenocarcinoma cell line LNCaP and in NIS-transfected myeloma cells, both of which exhibited prolonged retention of radio iodide even in the absence of I(-) organification. The therapeutic potential of alternative NIS-transported radioisotopes with different decay properties and a shorter, physical half-life than 131I(-), such as beta-emitter 188Rhenium (188ReO(4)-) and alpha-emitter 211Astatine (211At(-)), has been evaluated. In conclusion, it is clear that the remarkable progress made in the last few years in the molecular characterization of NIS has created new opportunities for the development of diagnostic and therapeutic applications for NIS in nuclear medicine. PMID- 14735457 TI - Imaging and therapy of tumors induced to express somatostatin receptor by gene transfer using radiolabeled peptides and single chain antibody constructs. AB - The fields of radioimmunodetection and radioimmunotherapy began with an initial paradigm that a targeting molecule (eg, antibody) carrying a radioisotope had the potential of selectively imaging and delivering a therapeutic dose of radiation to tumor sites. A second paradigm was developed in which injection of the targeting molecule was separated from injection of a short-lived radioisotope labeled ligand (so-called "pretargeting strategy"). This strategy has improved radioisotope delivery to tumors in animal models, enhanced radioimmune imaging in man, and therapeutic trials are in an early phase. We proposed a third paradigm to achieve radioisotopic localization at tumor sites by inducing tumor cells to synthesize a membrane expressed receptor with a high affinity for infused radiolabeled ligands. The use of gene transfer technology to induce expression of high affinity membrane receptors can enhance the specificity of radioligand localization, while the use of radioisotopes with the ability to deliver radiation damage across several cell diameters will compensate for less than perfect transduction efficiency. This approach was termed "Genetic Radioisotope Targeting Strategy." Using this strategy, induction of high levels of gastrin releasing peptide receptor or human somatostatin receptor subtype 2 expression and selective tumor uptake of radiolabeled peptides was achieved. The advantages of the genetic transduction approach are (1) constitutive expression of a tumor associated antigen/receptor is not required; (2) tumor cells are altered to express a new target receptor or increased quantities of an existing receptor at levels that may significantly improve tumor targeting of radiolabeled ligands compared with normal tissues; (3) gene transfer can be achieved by intratumoral or regional injection of gene vectors; (4) it is feasible to target adenovirus vectors to receptors overexpressed on tumor cells by modifying adenoviral tropism (binding) so that the virus will be targeted specifically to the desired tumor; and (5) it is possible to coexpress the receptor gene and a therapeutic gene, such as cytosine deaminase, for molecular prodrug therapy to produce an enhanced therapeutic effect. PMID- 14735458 TI - Molecular imaging in nuclear cardiology. AB - State-of-the-art techniques have been used to measure key aspects of cardiovascular pathophysiology from the birth of radionuclide cardiovascular imaging. However, during the last 30 years, there have been few innovative imaging advances to further our understanding of the complex physiologic processes. Molecular imaging now offers an array of tools to develop advanced diagnostic approaches and therapies for patients with coronary artery disease and heart failure. For example, the enhanced understanding of the pathophysiology of atheroma makes it possible to identify vulnerable plaque based on its metabolic signature or the presence of excessive apoptosis. Because the metabolic and apoptotic signals are large, it is likely that even small lesions will be visible. Of the many approaches that are being developed, 2 tracers appear most likely to be tested in the near future: (1) [18F]-fluorodeoxyglucose, to determine macrophage metabolism; and (2) radiolabeled annexin, to measure apoptosis of the inflammatory cells. Using existing techniques such as perfusion imaging, appropriate patients can be selected for treatment with novel therapies, such as stem cell transplantation or vascular gene therapy. Using positron tomography in place of single photon imaging adds the capability for the measurement of absolute perfusion and perfusion reserve to the information on regional perfusion. Flow reserve detects global decreases in perfusion and refines the determination of lesion severity available from perfusion imaging. PMID- 14735459 TI - Implications of PET based molecular imaging on the current and future practice of medicine. AB - The last quarter century has witnessed the introduction of a variety of powerful techniques that have allowed visualization of organ structure and function with exquisite detail. This in turn has brought about a true revolution in the day-to day practice of medicine. Structural imaging with x-ray computerized tomography and magnetic resonance imaging has added tremendously to many areas of medicine, including preoperative evaluation of patients. Many surgical procedures have been replaced by minimally invasive techniques, which have become a reality only because of the availability of modern imaging modalities. However, despite such accomplishments, structural imaging is quite insensitive for detecting early disease in which there often are no gross structural alterations in organ anatomy. Therefore, these modalities should be complemented by methodologies that can detect abnormalities at the molecular and cellular levels. The introduction of [(18)F]-fluorodeoxyglucose positron emission tomography (FDG-PET) in 1976 as a molecular imaging technique clearly has shown the power of this approach for treating a multitude of serious disorders. The impact of FDG-PET has been particularly impressive in patients with cancer diagnosis, for whom it has become important in staging, monitoring response to treatment, and detecting recurrence. In this review, we emphasize the role of FDG-PET in the assessment of central nervous system maladies, malignant neoplastic processes, infectious and inflammatory diseases, and cardiovascular disorders. New radiotracers are being developed and promise to expand further the list of indications for PET. These include novel tracers for cancer diagnosis and treatment capable of detecting hypoxia and angiogenesis. Prospects for developing new tracers for imaging other organ diseases also appear very promising. PMID- 14735460 TI - Neurochemical imaging of dementias. AB - Neurochemical imaging is one of the most established "molecular" imaging techniques. There have been tremendous efforts expended to develop radioligands specific to each neurochemical system. Investigational applications of neurochemical imaging in dementing disorders are extensive. Cholinergic, dopaminergic, and serotonergic systems, as well as benzodiazepine receptors, opioid receptors, and glutamatergic receptors have been imaged in Alzheimer disease and other dementing disorders. These investigations have provided important insights into disease processes in living human patients. The clinical diagnostic use of neurochemical imaging for dementing disorders is currently limited, but this technique is used to help develop therapeutic drugs at multiple levels. PMID- 14735461 TI - Enhancement of chemotherapeutic response of tumor cells by a heme oxygenase inhibitor, pegylated zinc protoporphyrin. AB - Heme oxygenase-1 (HO-1), an inducible enzyme that catalyzes oxidative degradation of heme to form biliverdin, carbon monoxide and free iron, may protect tumor cells against oxidative stress, thus contributing to rapid tumor growth in vivo. Here, we discuss whether pegylated zinc protoporphyrin (PEG-ZnPP), a potent HO inhibitor, modulates the chemotherapeutic response of tumor cells to treatment that generates reactive oxygen species (ROS). PEG-ZnPP is a water-soluble HO inhibitor that accumulates in tumor tissues after intravenous administration. Cytotoxicity of antitumor agents in vitro was determined by means of MTT and annexin V assays using human colon carcinoma SW480 cells. Mice bearing sarcoma 180 tumors were used as an in vivo model. Pegylated D-amino acid oxidase (PEG DAO), which behaves as an oxidative chemotherapeutic agent by generating toxic oxidants at tumor tissues, was administered with its substrate D-proline to mice with or without PEG-ZnPP pretreatment. PEG-ZnPP-treated SW480 cells became vulnerable to insults caused by various cytotoxic agents; the 50% lethal doses were reduced by 25%, 39%, 83%, and 61% for hydrogen peroxide, t-butyl hydroperoxide, camptothecin and doxorubicin, respectively. Cells treated with PEG ZnPP plus cytotoxic oxidants exhibited marked production of intracellular ROS, which paralleled the incidence of apoptosis. PEG-ZnPP pretreatment significantly reduced tumor growth in mice receiving PEG-DAO/D-proline compared to no PEG-ZnPP pretreatment. These findings suggest that HO-1 may become an attractive target for chemotherapeutic intervention. Further study of the effect of PEG-ZnPP plus conventional anticancer drugs that generate ROS, such as cisplatin, camptothecin, doxorubicin, mitomycin C and etoposide, is warranted. PMID- 14735462 TI - DNA polymerase theta is preferentially expressed in lymphoid tissues and upregulated in human cancers. AB - DNA polymerase theta (Pol theta) is a recently identified family A polymerase that contains an intrinsic helicase domain. Drosophila Pol theta mutants are hypersensitive to bifunctional DNA crosslinking agents and exhibit an elevated frequency of spontaneous chromosomal aberrations, suggesting a role for Pol theta in repair of DNA interstrand crosslinks and in the general maintenance of genome stability. To investigate a possible involvement of Pol theta in tumorigenesis, we have examined its expression in various normal and malignant tissues. Paired tumor and adjacent nontumorous tissues from patients with lung (n = 27), stomach (n = 28) and colon (n = 26) cancer, as well as 26 normal human tissues, were examined for Pol theta expression by RT-PCR, Northern or Western blot analysis. Pol theta was predominantly expressed in primary lymphoid organs including the fetal liver, thymus and bone marrow where lymphocyte progenitors undergo V(D)J rearrangements of their antigen receptor genes. In addition, Pol theta expression was upregulated in germinal center B cells, in which class switch recombination of the immunoglobulin genes occurs. Examination of Pol theta expression in matched cancer specimens revealed that Pol theta was barely detectable in the nontumorous tissues but was upregulated in 17 of 27 (63%) lung, 11 of 28 (39%) stomach and 20 of 26 (77%) colon cancers. Moreover, patients with high levels of Pol theta expression had a significantly poorer clinical outcome compared with those expressing low levels of Pol theta. These results implicate that Pol theta may have a specialized function in lymphocytes and that its overexpression may contribute to tumor progression. PMID- 14735463 TI - DNA bulky adducts in a Mediterranean population correlate with environmental ozone concentration, an indicator of photochemical smog. AB - Ozone (O(3)), the major oxidant component in photochemical smog, mostly derives from photolysis of nitrogen dioxide. O(3) may have biologic effects directly and/or via free radicals reacting with other primary pollutants and has been reported to influence daily mortality and to increase lung cancer risk. Although DNA damage may be caused by ozone itself, only other photochemical reaction products (as oxidised polycyclic aromatic hydrocarbons) may form bulky DNA adducts, a reliable biomarker of genotoxic damage and cancer risk, showing a seasonal trend. In a large series consisting of 320 residents in the metropolitan area of Florence, Italy, enrolled in a prospective study for the period 1993-1998 (206 randomly sampled volunteers, 114 traffic-exposed workers), we investigated the correlation between individual levels of DNA bulky adducts and a cumulative O(3) exposure score. The average O(3) concentrations were calculated for different time windows (0-5 to 0-90 days) prior to blood drawing for each participant, based on daily measurements provided by the local monitoring system. Significant correlations between DNA adduct levels and O3 cumulative exposure scores in the last 2-8 weeks before enrollment emerged in never smokers. Correlations were highest in the subgroup of never smokers residing in the urban area and not occupationally exposed to vehicle traffic pollution, with peak values for average concentrations 4-6 weeks before enrollment (r = 0.34). Our current findings indicate that DNA adduct formation may be modulated by individual characteristics and by the cumulative exposure to environmental levels of ozone in the last 4-6 weeks, possibly through ozone-associated reactive pollutants. PMID- 14735464 TI - Antisense abrogation of DENN expression induces apoptosis of leukemia cells in vitro, causes tumor regression in vivo and alters the transcription of genes involved in apoptosis and the cell cycle. AB - We previously reported that messenger RNA expression of DENN (differentially expressed in normal and neoplastic cells) is considerably higher in cancer cell lines than in normal cells. In our present study, we established that certain cancer cell lines express conspicuously higher levels of the 2 DENN isoforms in contrast to the 2 pro-apoptotic IG20 isoforms. Antisense DENN oligodeoxynucleotide treatment of K36 cells in vitro induced extensive apoptosis, while antisense DENN silencing of K36 tumor-bearing mice caused significant tumor regression in vivo. Compared to wild-type murine embryonic fibroblasts, antisense treatment of NFkappaB and TNFR1 KO cells resulted in markedly more pronounced cell death, whereas antisense-treated TNFalpha and TNFR2 knockouts exhibited less prominent apoptosis. Cell viability and apoptosis were authenticated by flow cytometry, membrane integrity, TUNEL, annexin V assays, histology and electron microscopy. Antisense abrogation of DENN expression culminated in upregulated expression of TNFR2, TRAIL and Fas, but downregulation of TNFalpha, TNFR1 and cyclin D3. Conversely, DENN overexpression stimulated cell proliferation and led to upregulated TRPM2 and cyclin B1, but diminished expression of Fas, TNFR2, TRAIL and Egr-1. The participation of TNFalpha, TNFR1, TNFR2 and Fas in the inhibition of DENN expression was also demonstrated. These data support the anti apoptotic and cell survival role of DENN, especially in malignant cells, and its interaction with specific genes and proteins involved in the apoptotic and cell cycle pathways. PMID- 14735465 TI - Bcl-xL inhibits p53- but not apoptin-induced apoptosis in head and neck squamous cell carcinoma cell line. AB - Nonfunctional p53 and especially upregulation of Bcl-x(L) result in advanced disease and poor prognosis of patients suffering head and neck squamous cell carcinoma (HNSCC). Aberrancies of Bcl-x(L) and/or p53 in HNSCC lead to inability of anticancer drugs to induce apoptosis. Bcl-x(L) and/or mutated p53 inhibit the apoptotic process by preventing the mitochondrial release of cytochrome c and/or activation of execution caspases. Here, we report that expression of the avian virus-derived apoptin protein resulted in induction of apoptosis in the HNSCC derived cell line UMSSC-14B despite the presence of nonfunctional p53. Apoptin activated the execution caspase 3 and induced the release of mitochondrial cytochrome c. Upregulation of Bcl-x(L) in UMSCC-14B cells did not interfere with the apoptin-induced apoptosis, whereas it clearly negatively affected the p53 induced one. Bcl-x(L) significantly decreased the p53-induced cytochrome c release, but not the apoptin-triggered one. Our data demonstrate that apoptin induces apoptosis independent of Bcl-x(L) and p53 and may constitute a potential therapeutic agent for treatment of HNSCC. PMID- 14735466 TI - Expression of uteroglobin in normal and carcinogenic endometrium and influence of hormone replacement therapy. AB - Uteroglobin, first reported in 1968 as a steroid secreted in rabbit uterine fluid during early pregnancy, is a progesterone-regulated and progesterone-binding protein. There is evidence that indicates that uteroglobin is inversely correlated to neoplastic growth but its role to endometrial carcinogenesis is not known. Therefore we analyzed the expression of uteroglobin in 13 normal endometrium, 19 hyperplasia and 21 endometrial carcinoma samples and the relation to estrogen receptor-alpha (ER-alpha) and progesterone receptor (PR) by immunohistochemistry and Western blotting. We also analyzed the expression of uteroglobin in 15 menopausal women who received hormone replacement therapy (HRT). The expression of uteroglobin was higher during the secretory phase than in the proliferative phase; however, it was detected in endometrial hyperplasia as weakly as in the proliferative phase and decreased according to the loss of differentiation in endometrial carcinoma. The results were basically in accord with those for PR; however, the expression of uteroglobin was weak, though PR was most detected in endometrial hyperplasia. In menopausal endometrium, the group treated with estrogen plus progesterone exhibited higher expression of uteroglobin than the group treated only with estrogen. The evidence suggests that uteroglobin expression is regulated by progesterone in the normal endometrium but that the regulation by PR is lost in endometrial hyperplasia and carcinoma according to acquirement of tumorigenesis and that estrogen plus progesterone therapy reduces the risk for endometrial carcinoma by restoring uteroglobin. PMID- 14735467 TI - Hyaluronan-binding peptide can inhibit tumor growth by interacting with Bcl-2. AB - Previous studies have indicated that proteins that bind hyaluronan can also inhibit the growth of tumor cells. To determine if synthetic peptides also possessed these properties, we tested a series of polypeptides containing structural motifs from different proteins for their ability to bind [(3)H]hyaluronan, and identified one compound termed P4 that had a particularly strong interaction. Further studies revealed that P4 also inhibited the growth of tumor cells in tissue culture as well as on the chorioallantoic membranes of chicken embryos. In addition, expression vectors for P4 caused tumor cells to grow slower in nude mice and reduced their vascularization. The P4 peptide also inhibited VEGF-induced angiogenesis in the chorioallantoic membranes of chicken embryos. Studies on cultured cells indicated that P4 induced apoptosis, which was blocked by a pan-caspase inhibitor. Confocal microscopy revealed that shortly after its uptake, P4 became associated with mitochondria. Immunoprecipitation indicated that P4 could bind to Bcl-2 and Bcl-x(L), which are associated with mitochondria and regulate apoptosis. This was also supported by the fact that P4 induced the release of cytochrome c from preparations of mitochondria. Taken together, these results suggest that P4 binds to Bcl-2 and related proteins and this activates the apoptotic cascade. PMID- 14735468 TI - Potent effect of 5-HPBR, a butanoate derivative of 4-HPR, on cell growth and apoptosis in cancer cells. AB - Fenretinide, 4-(N-hydroxyphenyl) retinamide (4-HPR), has demonstrated anticancer activity associated with a favorable toxicity profile and is now being investigated in several clinical trials. However, its plasma levels in patients have been far lower than the effective concentration required to induce apoptosis (usually 10 microM). This result has led to the synthesis of derivatives with better efficacy. Sodium butyrate's potential as an anticancer agent prompted us to synthesize a butanoate derivative of 4-HPR, 5-hydroxyphenyl butanoate retinamide (5-HPBR) and compare it to the parent compound for antitumor potential in vitro. The cytotoxicity of 5-HPBR was 2- to 6-fold greater than that of 4-HPR against cancer cell lines derived from various tissues. In premalignant bronchial cells (BEAS2B), 5-HPBR exhibited about a 10-fold stronger cytotoxicity than did 4 HPR. Normal CHANG liver cells were unaffected by either 4-HPR or 5-HPBR. Subsequent assays using DNA fragmentation, DAPI staining, FACS and Western blotting suggested that the potent inhibitory effect of 5-HPBR is mediated by apoptosis; the exact mechanism appears to differ among cancer cell types. In transcription assays with COS-1 cells, 5-HPBR selectively activated RARbeta and RARgamma but was a weaker ligand for all 3 subtypes of RAR than either all-trans retinoic acid or 4-HPR. Overall, these data suggest that 4-BHPR may be a promising retinoid with enhanced antitumor activity and reduced toxicity. PMID- 14735469 TI - Expression profile of genes associated with antimetastatic gene: nm23-mediated metastasis inhibition in breast carcinoma cells. AB - Metastases of various malignancies have been shown to be inversely related to the abundance of nm23 protein expression. However, the downstream pathways involved in nm23-mediated suppression of metastasis have not been elucidated. In the present investigation, we used cDNA microarrays to identify novel genes and functional pathways in nm23-mediated spontaneous breast metastasis. Microarray experiments were performed in a pair of cell lines, namely, C-100 (only vector transfected; highly metastatic) and H1-177 (nm23 transfected; low metastatic), derived from human mammary carcinoma cell line MDA-MB-435. The cDNA microarray analysis using GeneSpring software revealed significant as well as consistent alterations in the expression (up- and downregulation) of 2158 genes in a total of 18889 genes between high and low metastatic cells. Some of these genes were grouped into 6 functional categories, namely, invasion and metastasis, apoptosis and senescence, signal transduction molecules and transcription factors, cell cycle and repair, adhesion, and angiogenesis to extrapolate an association between these genes and different functional pathways involved in nm23-regulated metastasis. The results suggest that nm23 gene plays a major role in metastasis and its mechanism of action of metastasis suppression may involve downregulation of genes associated with cell adhesion, motility (integrins alpha2, -8, -9, -L and -V, collagen type VIII alpha1, fibronectin 1, catenin, TGF-beta2, FGF7, MMP14 and 16, ErbB2) and possibly certain tumor/metastasis suppressors (2 members of SWI/SNF-related matrix-associated proteins 2 and 5 and PTEN). PMID- 14735470 TI - Candidate regions of tumor suppressor locus on chromosome 9q31.1 in gastric cancer. AB - Loss of heterozygosity (LOH) is an important event of tumorigenesis. In gastric cancer, we found a novel region of LOH in chromosome 9q having about 800 kb deletions at 9q31.1. The microsatellite marker D9S938 in that region exhibiting the highest LOH frequency, 56.5%. In addition, the LOH at 9q31.1 did not show any relationship to either histologic types or stages of gastric cancers, and several genes were predicted in the remaining allele by in silico methods. These data suggest that the deletion at 9q31.1 would be common in both differentiated-type and undifferentiated-type gastric cancers. Furthermore, this deletion was found in the primary tumors of early-stage gastric cancer, indicating that loss of function of predicted genes appears to be associated with the tumorigenesis of gastric cancer. PMID- 14735471 TI - Parvovirus H-1 infection of human glioma cells leads to complete viral replication and efficient cell killing. AB - The extremely poor prognosis of malignant gliomas requires the investigation of other than standard therapies, i.e., the application of oncolytic viruses. In our study, we evaluated the effects of the oncosuppressive parvovirus H-1 on different established glioblastoma cell lines of rat and human origin and on short-term/low-passage cultures of human glioblastoma cells. We observed an efficient and dose-dependent killing of all glioma cell cultures at low multiplicities of infectious particles (MOI) per cell. Southern blot analysis of viral DNA amplification, RT-PCR analysis of viral RNA expression and Western blot analysis of the expression of viral structural (VP-1/VP-2) and nonstructural (NS 1) proteins demonstrated the biosynthesis of these viral macromolecular components in all of the cultures. Moreover, all the glioma cells were proficient for the production of infectious H-1 virus particles. The amount of virus production differed between a several fold increase of the input virus titer in most of the short-term/low-passage cultures up to 1,000-fold in one short-term glioma and in the rat cells. Glioma cells lines and, more importantly, short term/low-passage cultures of human glioblastomas were found to be highly susceptible target cells for H-1 virus mediated cytotoxicity. The formation of fully infectious progeny particles in infected glioma cells offers the chance for the induction of secondary rounds of infection resulting in an advanced cytotoxic effect. These advantageous characteristics of H-1 virus infection of glioma cells, combined with the known low toxicity of H-1 virus in nontransformed cells, make parvovirus H-1 a promising candidate for oncolytic glioma therapy. PMID- 14735472 TI - Targeting and amplification of immune killing of tumor cells by pro-Smac. AB - Overexpression of inhibitors of apoptosis (IAP) is one potential mechanism for tumor cells to evade immune surveillance. To determine whether immune-mediated killing of tumor cells can be enhanced by neutralization of IAP proteins, 2 novel eGFP-Smac fusion proteins (pro-Smac) were introduced into the poorly immunogenic mouse melanoma cell line, B16BL6-D5 (D5). Each fusion protein contained Smac and a cleavage site specific for granzyme B (GrB) or caspase 8, thereby targeting the 2 major killing mechanisms of cytotoxic T-lymphocyte (CTL) and NK cells. Expression of a pro-Smac fusion protein by D5 tumor cells greatly enhanced the susceptibility to killing by lymphokine-activated killer (LAK) cells or purified GrB. GrB-mediated killing was increased to a much greater extent when tumor cells expressed the eGFP-Smac fusion protein with a GrB cleavage site compared to a caspase 8 cleavage site. In contrast, perforin-deficient LAK cells, which lack GrB-mediated cytotoxicity but process normal ligands for death receptors, killed D5 tumor cells expressed pro-Smac with caspase 8 cleavage site more efficiently. Enhanced killing by GrB was also accompanied by processing of the fusion protein and increased caspase-3-like activity. These results indicate that killing of tumor cells can be amplified by targeting cell-mediated cytotoxic mechanisms via expression of pro-Smac fusion proteins. PMID- 14735473 TI - Glutathione S-transferase M3 (A/A) genotype as a risk factor for oral cancer and leukoplakia among Indian tobacco smokers. AB - Polymorphism in glutathione S-transferase (GST) genes, causing variations in enzyme activities, may influence susceptibility to oral cancer and leukoplakia in smokers and/or smokeless tobacco users. In this case-control study consisting of 109 leukoplakia and 256 oral cancer patients and 259 controls, genotype frequencies at GSTM1, GSTT1, GSTM3 and GSTP1 loci were determined by polymerase chain reaction-restriction fragment length polymorphism methods and analyzed by multiple logistic regression to determine the risks of the diseases. There were no significant differences in the distributions of GSTM1, GSTM3 and GSTT1 genotypes in patients and controls when all individuals were compared. In contrast, frequencies of ile/ile genotype at codon 105 and variant val-ala haplotype of GSTP1 was significantly higher (OR = 1.5; 95% CI = 1.0-2.0) and lower (OR = 1.4; 95% CI = 1.0-1.9) in oral cancer patients compare to controls, respectively. The impacts of all genotypes on risks of oral cancer and leukoplakia were also analyzed in patients with different tobacco habits and doses. Increased risks of cancer and leukoplakia were observed in tobacco smokers with GSTM3 (A/A) genotype (OR = 2.0, 95% CI = 1.0-4.0; OR = 2.0, 95% CI = 1.0 4.4, respectively). So, GSTM3 (A/A) genotype could become one of the markers to know which of the leukoplakia would be transformed into cancer. Heavy tobacco chewing (> 124 chewing-year) increased the risk of cancer in individuals with GSTT1 homozygous null genotype (OR = 3.0; 95% CI = 1.0-9.8). Furthermore, increased lifetime exposure to tobacco smoking (> 11.5 pack-year) increased the risk of leukoplakia in individuals with GSTM1 homozygous null genotype (OR = 2.4; 95% CI = 1.0-5.7). It may be suggested that polymorphisms in GSTP1, GSTM1, GSTM3 and GSTT1 genes regulate risk of cancer and leukoplakia differentially among different tobacco habituals. PMID- 14735474 TI - Cyclin E expression in papillary thyroid carcinoma: relation to staging. AB - Cyclin E plays a pivotal role in the regulation of G1-S transition and relates to malignant transformation of the cells. However, the clinical significance of cyclin E expression in patients with papillary thyroid carcinoma (PTC) remains unknown. We examined by immunohistochemistry the expression of cyclin E in 41 resected PTCs in pathologic stages from pT1a to pT4 and analyzed its relation to clinicohistopathologic factors. The positive staining was divided into 3 grades: no expression if less than 10%, expression if 11-50% and overexpression if more than 50% of the nuclei of tumor cells were stained positively. Cylin E expressions were observed in 75.6% of analyzed PTCs but only 60% of papillary microcarcinomas (PMCs) were immunopositive for cyclin E expression. However, cyclin E staining was observed in 90.4% of PTCs in a group with TNM higher than pT1a. The staining index was significantly different between the PMCs and the rest of the cancers investigated (14.91% +/- 14.4% vs. 34.03% +/- 23.44%, respectively; p < 0.005) and we observed positive relation between the staining index and factor T of staging of PTCs. All the lymph node metastases coexisted with cyclin E expression and most, but not all, of them coexisted with cyclin E overexpression. These findings indicate that cyclin E may play a key role for the oncogenesis and biologic behavior of PTC. If our results are confirmed in a larger study, a high level of cyclin E expression may become a new prognostic marker for PTCs. PMID- 14735475 TI - Expression of LDL receptor-related protein 5 (LRP5) as a novel marker for disease progression in high-grade osteosarcoma. AB - The Wingless-type (Wnt) family of proteins and its coreceptor LRP5 have recently been implicated in human skeletal development. Wnt pathway modulates cell fate and cell proliferation during embryonic development and carcinogenesis through activation of receptor-mediated signaling. Osteosarcoma (OS) is a bone-forming tumor of mesenchymal origin whose growth control has been linked to autocrine or paracrine stimulation by several growth factor families. We examined 4 OS cell lines for WNT1, WNT4, WNT5A, WNT7A, WNT11, FZD1-10 and LRP5 expression by reverse transcription polymerase chain reaction (RT-PCR). In addition, RT-PCR for LRP5 expression was performed in 44 OS patient samples and the findings were correlated with clinical data. Expression profiling of Wnts and their receptors revealed the presence of several isoforms in OS cell lines. Overall, 22/44 (50%) of OS patient samples showed evidence of LRP5 expression. Presence of LRP5 correlated significantly with tumor metastasis (p = 0.005) and the chondroblastic subtype of OS (p = 0.045). In addition, patients whose tumors were positive for LRP5 showed a trend toward decreased event-free survival (p = 0.066). No significant association was found between LRP5 expression and age, gender, site of disease, site of metastasis or degree of chemotherapy-induced tumor necrosis. Sequencing of exon 3 of LRP5 in 10 OS patient-derived cell cultures showed no activating mutation of LRP5. These results showed that expression of LRP5 is a common event in OS and strongly suggest a role for LRP and Wnt signaling in the pathobiology and progression of this disease. PMID- 14735476 TI - Risk factors for squamous intraepithelial lesions (SIL) of the cervix among women residing at the US-Mexico border. AB - It is now well established that cervical cancer is caused by oncogenic human papillomavirus (HPV) infections that commonly infect women worldwide. What remains to be understood are the factors that contribute to cervical cancer in the presence of HPV infection. We conducted a case-control analysis of women recruited at the US-Mexico border to simultaneously evaluate factors associated with 3 cytologic outcomes: atypical squamous cells of undetermined significance or atypical glandular cells of undetermined significance (ASCUS/AGUS), low grade squamous intraepithelial lesions (LSIL) and high grade squamous intraepithelial lesions (HSIL). A cross-sectional binational study of 2059 women ages 15-79 years was conducted between 1997 and 1998. A significant difference in the distribution of cytologic categories by country was observed (3.0% vs. 0.7% HSIL among Mexican vs. US women, respectively). The only factors independently associated with all 3 cytologic outcomes were HPV infection and viral load. A linear increase in risk with increasing viral load was observed for each of the 3 outcome variables, with the strength of this association increasing with cytology abnormality. In addition to HPV infection, parity and Mexico as a country of residence appear to be associated with LSIL and HSIL, respectively. Factors associated with cytologic outcomes in analyses limited to women with HPV infection were similar to results obtained in models where HPV infection was included as a covariate. Future work is needed to evaluate the predictive value of HPV viral load utilizing more specific and quantitative measures. PMID- 14735477 TI - Fruits and vegetables and prostate cancer: no association among 1104 cases in a prospective study of 130544 men in the European Prospective Investigation into Cancer and Nutrition (EPIC). AB - We examined the association between self-reported consumption of fruits and vegetables and prostate cancer risk in the European Prospective Investigation into Cancer and Nutrition (EPIC). Data on food consumption and complete follow-up for cancer incidence were available for 130544 men in 7 countries recruited into EPIC between 1993 and 1999. After an average of 4.8 years of follow-up, there were 1104 incident cases of prostate cancer. The associations of consumption of total fruits, total vegetables, cruciferous vegetables and combined total fruits and vegetables with prostate cancer risk were examined using Cox regression, stratified for recruitment center and adjusted for height, weight and energy intake. There was a wide range in consumption of fruits and vegetables: mean intakes (g/day) in the bottom and top fifths of the distribution, as estimated from 24-hr recalls in a subsample of participants, were 53.2 and 410.7 for fruits, 97.1 and 242.1 for vegetables and 169.0 and 633.7 for fruits and vegetables combined. No significant associations between fruit and vegetable consumption and prostate cancer risk were observed. Relative risks (95% confidence intervals) in the top fifth of the distribution of consumption, compared to the bottom fifth, were 1.06 (0.84-1.34) for total fruits, 1.00 (0.81 1.22) for total vegetables and 1.00 (0.79-1.26) for total fruits and vegetables combined; intake of cruciferous vegetables was not associated with risk. These results suggest that total consumption of fruits and vegetables is not associated with the risk for prostate cancer. PMID- 14735478 TI - Secondhand smoke exposure in adulthood and risk of lung cancer among never smokers: a pooled analysis of two large studies. AB - The interpretation of the evidence linking exposure to secondhand smoke with lung cancer is constrained by the imprecision of risk estimates. The objective of the study was to obtain precise and valid estimates of the risk of lung cancer in never smokers following exposure to secondhand smoke, including adjustment for potential confounders and exposure misclassification. Pooled analysis of data from 2 previously reported large case-control studies was used. Subjects included 1263 never smoking lung cancer patients and 2740 population and hospital controls recruited during 1985-1994 from 5 metropolitan areas in the United States, 11 areas in Germany, Italy, Sweden, United Kingdom, France, Spain and Portugal. Odds ratios (ORs) of lung cancer were calculated for ever exposure and duration of exposure to secondhand smoke from spouse, workplace and social sources. The OR for ever exposure to spousal smoking was 1.18 (95% CI = 1.01-1.37) and for long term exposure was 1.23 (95% CI = 1.01-1.51). After exclusion of proxy interviews, the OR for ever exposure from the workplace was 1.16 (95% CI = 0.99-1.36) and for long-term exposure was 1.27 (95% CI = 1.03-1.57). Similar results were obtained for exposure from social settings and for exposure from combined sources. A dose response relationship was present with increasing duration of exposure to secondhand smoke for all 3 sources, with an OR of 1.32 (95% CI = 1.10-1.79) for the long-term exposure from all sources. There was no evidence of confounding by employment in high-risk occupations, education or low vegetable intake. Sensitivity analysis for the effects of misclassification (both positive and negative) indicated that the observed risks are likely to underestimate the true risk. Clear dose-response relationships consistent with a causal association were observed between exposure to secondhand smoke from spousal, workplace and social sources and the development of lung cancer among never smokers. PMID- 14735479 TI - Risk of lung cancer and residential radon in China: pooled results of two studies. AB - Studies of radon-exposed underground miners predict that residential radon is the second leading cause of lung cancer mortality; however, case-control studies of residential radon have not provided unambiguous evidence of an association. Owing to small expected risks from residential radon and uncertainties in dosimetry, large studies or pooling of multiple studies are needed to fully evaluate effects. We pooled data from 2 case-control studies of residential radon representing 2 large radon studies conducted in China. The studies included 1050 lung cancer cases and 1996 controls. In the pooled data, odds ratios (OR) increased significantly with greater radon concentration. Based on a linear model, the OR with 95% confidence intervals (CI) at 100 Becquerel/cubic-meter (Bq/m(3)) was 1.33 (1.01,1.36). For subjects resident in the current home for 30 years or more, the OR at 100 Bq/m(3) was 1.32 (1.07,1.91). Results across studies were consistent with homogeneity. Estimates of ORs were similar to extrapolations from miner data and consistent with published residential radon studies in North American and Europe, suggesting long-term radon exposure at concentrations found in many homes increases lung cancer risk. PMID- 14735480 TI - Progression of chronic atrophic gastritis associated with Helicobacter pylori infection increases risk of gastric cancer. AB - We conducted a longitudinal cohort study to determine the association of Helicobacter pylori infection and the progression of chronic atrophic gastritis (CAG) with gastric cancer. A cohort of 4655 healthy asymptomatic subjects was followed for a mean period of 7.7 years. H. pylori infection was established by serum specific antibodies and the presence of CAG was confirmed by serum pepsinogen. During the follow-up period, 45 gastric cancer cases were detected (incidence rate, 126/100000 person-years). A univariate analysis after adjustment for age showed that both H. pylori and CAG were significantly associated with gastric cancer. To clarify the interaction between H. pylori and CAG, an analysis stratified by H. pylori- and CAG-status was performed. No cancer developed in the H. pylori(-)/CAG(-) group during the study period. This supports the theory that it is quite rare for any type of gastric cancer to develop in an H. pylori-free healthy stomach. With the progression of H. pylori-induced gastritis, the risk of gastric cancer increased in a stepwise fashion from CAG-free gastritis [H. pylori(+)/CAG(-) group] (HR=7.13, 95%CI=0.95-53.33) to CAG [H. pylori(+)/CAG(+) group] (HR=14.85, 95%CI=1.96-107.7) and finally to severe CAG with extensive intestinal metaplasia [H. pylori(-)/CAG(+) group] (HR=61.85, 95%CI=5.6-682.64) in which loss of H. pylori from the stomach is observed. Therefore, it is probable that H. pylori alone is not directly associated with stomach carcinogenesis. Instead, H. pylori appears to influence stomach carcinogenesis through the development of CAG. The observed positive correlation between the extent of H. pylori-induced gastritis and the development of cancer was strong, especially for the intestinal type. These results are compelling evidence that severe gastritis with extensive intestinal metaplasia is a major risk factor for gastric cancer, and they confirm the previously described model of stomach carcinogenesis: the gastritis-metaplasia-carcinoma sequence. PMID- 14735481 TI - Prognosis of dermal lymphatic invasion with or without clinical signs of inflammatory breast cancer. AB - It is still an open debate whether tumor emboli in dermal lymphatics without inflammatory signs represent a similar bad prognosis like inflammatory breast cancer. We evaluated the prognostic role of dermal lymphatic invasion (DLI) in breast cancer with (DLI + ID) or without (DLI w/o ID) inflammatory disease (ID). From August 1988 to January 2000, 42 patients with DLI were irradiated. Twenty five were classified as pT4, 13 out of them as pT4d (inflammatory disease); the 17 remaining patients had 1 T1c, 12 T2 and 4 T3 cancers with DLI. Axillary dissection revealed node-positive disease in 39/41 patients (median, 9 positive nodes). Thirty-eight out of 42 patients received adjuvant systemic treatment(s). After a mean follow-up of 33 months, 22/42 patients (52%) are disease-free. The actuarial 3-year disease-free survival is 50% (DLI w/o ID, 61%; DLI + ID, 31%; p < 0.03); the corresponding overall survival was 69% (DLI w/o ID, 87%; DLI + ID, 37%; p = 0.005). The presence or absence of ID was the only significant parameter for all endpoints in multivariate analyses. Dissemination occurred in 19 (45%), local relapse in 7 (n = 17%) and regional failure in 4 (10%). Nine patients (21%) had contralateral breast cancer/relapse. Despite the same histopathologic presentation, DLI w/o ID offered a significantly better disease-free survival and overall survival than ID. The finding of dermal lymphatic tumor invasion predicts a high probability for node-positive disease. PMID- 14735482 TI - Obesity and colon cancer: does leptin provide a link? AB - Obesity, a risk factor for colorectal cancer, is associated with elevated serum levels of leptin, the adipocyte-derived hormone, and insulin. Experimental and epidemiologic studies have indicated a role for insulin in the pathogenesis of colon cancer, and recent experimental studies have suggested a similar role for leptin. In a case-control study nested in the Janus Biobank, Norway, we measured serum levels of leptin and C-peptide (a marker of pancreatic insulin secretion) in cryopreserved prediagnostic sera from men (median age, 45 years) who were diagnosed with cancer of the colon (n = 235) or rectum (n = 143) after blood collection (median time, 17 years), and among 378 controls matched for age and date of blood collection. Conditional logistic regression analyses showed an approximately 3-fold increase in colon cancer risk with increasing concentrations of leptin up to an odds ratio (OR) of 2.72 (95% CI = 1.44-5.12) for top vs. bottom quartile (p(trend) = 0.008). The corresponding OR for C-peptide was 1.81 (95% CI = 0.67-4.86; p(trend) = 0.19). The risk estimates remained unchanged after mutual adjustment. No association of hormone levels with rectal cancer risk was found. Reproducibility of hormone measurements assessed by intraclass coefficients (ICCs) for paired samples taken 1 year apart was high for leptin (ICC = 0.82) but lower for C-peptide (ICC = 0.30). Our results suggest that leptin is a risk factor for colon cancer, and that leptin may provide a link between obesity and colon cancer. Leptin may be directly involved in colon tumorigenesis or it may serve as a sensitive and robust marker of an obesity induced adverse endocrine environment. Only weak support for an association of insulin with colon cancer was found. PMID- 14735483 TI - Inequalities in the healthcare system and breast cancer survival. PMID- 14735485 TI - Dominant expression of porcine Calbindin-D9k in the uterus during a luteal phase. AB - Calbindin-D9k (CaBP-9k) is a member of intracellular calcium binding proteins, which have a high affinity to calcium. CaBP-9k is mainly expressed in the mammalian intestine, uterus and placenta, and is regulated in tissue- and species specific manners. Previous studies have shown that CaBP-9k expression is mainly controlled by steroid hormones and their receptors. Thus, we further investigated the expression and regulation of CaBP-9k during an estrus cycle in the pig uterus by Northern blot and immunoblot analysis in this study. In addition, serum levels of estrogen (E2) and progesterone (P4) were measured using ELISA. The CaBP-9k mRNA is highly expressed in the porcine uterus during a luteal phase compared to a follicular phase, and its mRNA level in a luteal phase is increased up to 10 fold compared to a follicular phase. In parallel to the level of CaBP-9k mRNA, the CaBP-9k protein is also dominantly expressed in the porcine uterus, and strongly expressed in the epithelium and glands of the porcine uterus during a luteal phase. Although, the localization of the CaBP-9k protein is scarcely detected at follicular phase, it is dominantly expressed in the porcine uterus during a luteal phase. In addition, the serum P4 level was significantly increased during a luteal phase compared to a follicular phase, whereas no difference was observed in E2 levels between follicular and luteal phases, indicating that the ratio of P4/E2 is remarkably increased in porcine uterus during a luteal phase compared to a follicular phase. In conclusion, these results suggest that P4 may play an important role in the up-regulation of CaBP 9k gene in the porcine uterus in a luteal phase, which is unlike the condition in the rat uterus. In addition, the porcine CaBP-9k may be dominantly expressed in the epithelium and glandular structure of pig uterus during a luteal phase. It may also be differentially regulated during this cycle presumably by steroid hormones, especially up-regulated P4 levels in this tissue. PMID- 14735486 TI - Quantitative analysis throughout pregnancy of placentomal and interplacentomal expression of pregnancy-associated glycoproteins-1 and -9 in the cow. AB - The multigenic pregnancy-associated glycoproteins (PAG) exhibit spatially and temporally distinct pattern across gestation in the bovine. The majority of the bovine bPAG are localized to the binucleate cells (BNC) while some are expressed throughout the trophectoderm. Bovine (b)PAG-1 and -9 are both localized to the BNC but are differentially transcribed. In addition, the anatomical location of BNC does influence protein expression in the ungulates. Therefore, the objective of the present study was to compare and contrast bPAG-1 and -9 transcriptions in the placentomal (cotyledonary, caruncular) and interplacentomal (intercotyledonary, intercaruncular) tissues throughout pregnancy in the bovine using real-time reverse transcription PCR (RT-PCR) and by in-situ hybridization. The levels of bPAG-9 transcription in the fetal membrane at peri-implantation were significantly (P<0.01) higher than bPAG-1. The expression of bPAG-9 in the placentomal and interplacentomal tissues were significantly (P<0.01) higher than bPAG-1 during the first trimester of gestation. The transcription of bPAG-1 in the placentomal and interplacentomal tissues were significantly (P<0.01) higher than bPAG-9 from mid-gestation to peripartum. The expression of bPAG-1 and -9 throughout gestation were significantly (P<0.01) affected by the anatomical location of BNC. In situ analysis paralleled the expression patterns of bPAG-1 and -9 across gestation. These findings indicate that bPAG-9 expression in the placentomal and interplacentomal tissues predominates in the first trimester of gestation while bPAG-1 transcription was primarily higher in the last two trimesters of gestation. The cellular location had significant effect on bPAG-1 and -9 transcription. PMID- 14735487 TI - Stage-specific and tissue-specific expression characteristics of differentially expressed genes during mouse spermatogenesis. AB - Spermatogenesis occurs in successive mitotic, meiotic, and post-meiotic phase, and involves a number of unique processes including meiosis and dramatic morphological changes. The unique differentiation mechanisms of spermatogenesis suggest the existence of germ-cell-specific molecules. The most straight forward strategy to elucidate differentiation mechanisms is to identify and characterize differentiation-specific molecules and their associated genes in germ cells. However, only a few genes specifically involved in spermatogenesis have been studied. In the present study, six different types of spermatogenic cells (primitive type A spermatogonia, type B spermatogonia, preleptotene spermatocytes, pachytene spermatocytes, round spermatids, and elongating spermatids) were isolated from Balb/c mice testes using velocity sedimentation and Atlas cDNA arrays containing 1,176 known mouse genes were used to determine the gene expression profiles of the spermatogenic cells. The expression of 260 genes were detected in six different stages of spermatogenic cells and a number of genes showed differential expression. The 23 differentially expressed genes were further analysed by reverse transcription polymerase chain reaction (RT-PCR) for their stage-specific and tissue-specific expression characteristics. Based on the results of RT-PCR, six genes highly express in both primitive type A and type B spermatogonia, four genes up-regulate in type B spermatogonia, two genes up regulate in spermatocytes, two genes up-regulate in spermatids, three genes express constantly from primitive A spermatogonia to elongating spermatids, two genes express constantly from primitive A spermatogonia to round spermatids, two genes do not change in their expression during spermatogenesis, two genes can be detected highly in adult testis, but are undetectable in spermatogenic cells. The tissue-specific expression characteristics of the 23 genes showed that some of them specifically expressed in testes or other tissues. These data provide new information for further studies into spermatogenesis-related genes and may lead to the identification of genes with potential relevance to the differentiation of spermatogenic cells. In addition, some of these genes could be considered to be used as the molecular markers for different stages of spermatogenic cells. PMID- 14735488 TI - Molecular cloning and expression analysis of vitellogenin in scallop, Patinopecten yessoensis (Bivalvia, Mollusca). AB - The present study was undertaken to determine scallop vitellogenin (Vtg) cDNA sequence, to identify Vtg synthesizing cell, and to analyze the regulation of Vtg mRNA expression. Clones containing partial cDNA sequence of Vtg were isolated from cDNA library of the scallop ovary by immunoscreening with the anti-scallop vitellin (Vn) serum. The deduced amino acid sequence of the clone containing the longest cDNA insert (1,689 bp) was identified as a member of the lipid transport protein family and exhibited about 20-35% identity with Vtgs of other oviparous animals. Northern blot analysis identified a single transcript longer than 10 kb in the ovary. Dot blot analysis of the ovary showed a high amount of Vtg mRNA during the growing stage and the level was retained until spawning stage. In situ hybridization demonstrated the expression of Vtg mRNA in the auxiliary cells closely associated with growing oocytes, suggesting that the synthesis of a major yolk protein in the scallop occurs through hetero-synthetic pathway without mediation through the blood flow but occurs de novo in the ovary. The content of Vtg mRNA in the ovarian tissue cultured in vitro with vitellogenesis promoting factor (VPF), which strongly promotes Vtg protein synthesis, from the cerebral plus pedal ganglion (CPG) showed no change. The transcription of Vtg mRNA appeared to be promoted by estradiol-17beta (E2) not by VPF although VPF may enhance the translation of Vtg mRNA. PMID- 14735489 TI - Ovarian cysteine proteinases in the teleost Fundulus heteroclitus: molecular cloning and gene expression during vitellogenesis and oocyte maturation. AB - The cysteine proteinases cathepsins B and L are members of the multigene family of lysosomal proteases that have been implicated in the processing of yolk proteins (YPs) in teleost oocytes. However, the full identification of the type of cathepsins expressed in fish ovarian follicles and embryos, as well as their regulatory mechanisms and specific function(s), are not yet elucidated. In this study, cDNAs encoding cathepsins B, L, F, K, S, Z, C, and H have been isolated from the teleost Fundulus heteroclitus, and the analysis of their deduced amino acid sequences revealed highly similar structural features to vertebrate orthologs, and confirmed in this species the existence of cathepsin L-like, cathepsin B-like, and cathepsin F-like subfamilies of cysteine proteinases. While all identified cathepsins were expressed in ovarian follicles, the corresponding mRNAs showed different temporal expression patterns. Thus, similar mRNA levels of cathepsins L, F, S, B, C, and Z were found throughout the oocyte growth or vitellogenesis period, whereas those for cathepsin H and K appeared to decrease as vitellogenesis advanced. During oocyte maturation, a transient accumulation of cathepsins L, S, H, and F mRNAs, approximately a 3-, 1.5-, 1.6-, and 6-fold increase, respectively, was detected in ovarian follicles within the 20-25 hr after hormone stimulation, coincident with the maximum proteolysis of the oocyte major YPs. The specific temporal pattern of expression of these genes may indicate a potential role of cathepsin L-like and cathepsin F proteases in the YP processing events occurring during fish oocyte maturation and/or early embryogenesis. PMID- 14735490 TI - Tektin3 encodes an evolutionarily conserved putative testicular microtubules related protein expressed preferentially in male germ cells. AB - Tektins are microtubule-associated cytoskeletal proteins that are expressed primarily in the male germ cell-lineage in centrioles and basal bodies and within ciliary and flagellar doublet microtubules. They are proposed to be important for axonemal architecture and microtubule stability in the sperm tail and in other ciliated and flagellar structures. Using an in silico (electronic database) subtractive approach to identify germ cell-specific genes in vertebrates, we isolated a new member of the Tektin gene family from mice, Tektin3. Reverse transcriptase-polymerase chain reaction (RT-PCR) and Northern blot analyses confirm that Tektin3 encodes a 1.7 kb transcript detectable preferentially in the testes of adult mice. In situ hybridization analysis in the testes revealed Tektin3 mRNA expression exclusively in late pachytene spermatocytes and early round spermatids. The putative TEKTIN3 protein shares 83.5% overall sequence identity with the human ortholog and includes a fully conserved carboxy terminal nonapeptide signature sequence present in all TEKTIN family members. In addition, using database mining, highly conserved TEKTIN3 orthologs were identified in puffer fish (Fugu rubripes) and rats (Rattus norvegicus) which shared 60.9 and 91.4% identity, respectively, with mouse TEKTIN3. The Tekt3 gene maps to murine chromosome 11 in a region that is syntenic to the human 17p12 chromosomal region containing the human TEKTIN3 gene. Our studies demonstrate that TEKTIN3 is a novel evolutionarily conserved male germ cell-enriched protein and suggest that it might perform important roles in male reproductive development and physiology. PMID- 14735491 TI - Aromatase inhibitor induces complete sex change in the protogynous honeycomb grouper (Epinephelus merra). AB - The protogynous hermaphrodite fish change sex from female to male at the certain stages of life cycle. The endocrine mechanisms involved in gonadal restructuring throughout protogynous sex change are not clearly understood. In the present study, we implanted maturing female honeycomb groupers with nonsteroidal aromatase inhibitor (AI), Fadrozole (0, 1, and 10 mg/fish) and examined changes in gonadal structures and serum levels of sex steroid hormones 2(1/2) months after implantation. The ovaries of control females had oocytes undergoing active vitellogenesis, whereas AI caused females to develop into functional males. These males had testes, which were indistinguishable in structure from those of normal males, but bigger in size, and completed all stages of spermatogenesis including accumulation of large amount of sperm in the seminiferous tubules. AI significantly reduced the serum levels of estradiol-17beta (E2) and increased levels of testosterone (T), 11-ketotestosterone (11-KT), and 17alpha, 20beta dihydroxy-4-pregnen-3-one (DHP). Further, AI suppressed in vitro production of E2, and stimulated the production of T and 11-KT in the ovarian fragments of mature female. In the honeycomb grouper, suppression of both in vitro and in vivo production of E2 and degeneration of oocytes by AI suggests that AI induces complete sex change through inhibition of estrogen biosynthesis, and perhaps, subsequent induction of androgen function. PMID- 14735492 TI - Enhancement of prostacyclin synthesis at the beginning of formation of caprine corpora lutea. AB - We examined changes in the levels of prostacyclin (PGI2) synthase mRNA and 6-keto PGF1alpha, a stable metabolite of PGI2, in the caprine corpus luteum (CL) during its development and subsequent maintenance. We also looked at the effects of a potent GnRH antagonist (GA), which is known to suppress the release of luteinizing hormone (LH), on the PGI2 synthase mRNA level and the 6-keto PGF1alpha content during CL development. Goats were divided into a control group (n=12) and a GA-treated group (n=6). They were treated with saline or GA (50 microg/kg, s.c.) on days 0 (day of ovulation), 4, and 8 (control only), and the CL were collected from a subset of goats (n=3 for each day) on days 0 (no saline), 4, 8, or 14 (control only). Ribonuclease protection assay was performed to quantify the mRNA in the CL using specific cRNA probes generated by RT-PCR and in vitro transcription. The 6-keto-PGF1alpha content in the CL was measured by radioimmunoassay. The level of PGI2 synthase mRNA and the 6-keto-PGF1alpha content in the CL in the control group decreased from day 0 to day 4 (P<0.01), and did not change thereafter from day 4 to day 14. Levels of PGI2 synthase mRNA and 6-keto-PGF1alpha content in the CL on days 4 and 8 were not affected by treatment with GA. These results suggest that PGI2 synthesis is regulated upward at the beginning of caprine CL formation; this may play a role in initiating CL development. This study also suggests that changes of PGI2 synthesis during CL development are probably not regulated by LH. PMID- 14735493 TI - Study of newly synthesized proteins during bovine oocyte maturation in vitro using image analysis of two-dimensional gel electrophoresis. AB - To investigate protein synthesis during bovine oocyte maturation in vitro, oocytes were put in culture with 35S-methionine for 4 hr periods from time zero to 28 hr. Pools of 10 oocytes were then prepared for two-dimensional gel electrophoresis (2-DE). For each time interval, three gels were obtained, digitalized, and analyzed to detect proteins. Then, the gel containing the most proteins was chosen as the reference gel and compared with the others. An averaged gel was created with proteins present in at least two gels of the three. Our results indicate that the rate of protein synthesis is higher at the beginning of maturation until the appearance of metaphase I (MI, 8-12 hr) and then it decreases and stays relatively constant. Percentages of initial proteins (0-4 hr) and remaining present during the progression decrease progressively from 100% to 53%. In contrast, when we compare proteins synthesized from the 4 to 8 hr period with proteins from the 8 to 12 or the 12 to 16 hr intervals, percentages of overall protein matching are stable with values of 81 and 79%, respectively. Comparison of proteins from 20 to 24 hr with proteins from 16 to 20 or 24 to 28 hr intervals also gives stable percentages of overall protein matching with values of 83 and 84%, respectively. Furthermore, a higher number of new proteins is observed at 4-8 hr (n=130) and 16-20 hr (n=136) of maturation. Thus, three major patterns of protein synthesis were observed during bovine oocyte maturation in vitro: one at the beginning of maturation (0-4 hr), another one in the middle (4-16 hr), and the last one after the completion of MI stage (16-28 hr). PMID- 14735494 TI - Expression of mRNAs for DNA methyltransferases and methyl-CpG-binding proteins in the human female germ line, preimplantation embryos, and embryonic stem cells. AB - Recent evidence indicates that mammalian gametogenesis and preimplantation development may be adversely affected by both assisted reproductive and stem cell technologies. Thus, a better understanding of the developmental regulation of the underlying epigenetic processes that include DNA methylation is required. We have, therefore, monitored the expression, by PCR, of the mRNAs of DNA methyltransferases (DNMTs), methyl-CpG-binding domain proteins (MBDs), and CpG binding protein (CGBP) in a developmental series of amplified cDNA samples derived from staged human ovarian follicles, oocytes, preimplantation embryos, human embryonic stem (hES) cells and in similar murine cDNA samples. Transcripts of these genes were detected in human ovarian follicles (DNMT3A, DNMT3b1, DNMT3b4, DNMT1, MDBs1-4, MeCP2, CGBP), germinal vesicle (GV) oocytes (DNMT3A, DNMT3b1, DNMT1, MDBs1-4, MeCP2, CGBP), mature oocytes (DNMT3A, DNMT3b1, DNMT1, CGBP), and preimplantation embryos (DNMT3A, DNMT3b1, DNMT1, DNMT3L, MBD2, MDB4, CGBP). Differential expression of DNMT3B gene transcripts in undifferentiated (DNMT3b1) and in vitro differentiated human ES cells (DNMT3b3) further demonstrated an association of the DNMT3b1 transcript variant with totipotent and pluripotent human cells. Significantly, whilst the murine Dnmt3L gene is both expressed and essential for imprint establishment during murine oogenesis, transcripts of the human DNMT3L gene were only detected after fertilisation. Therefore, the mechanisms and/or the timing of imprint establishment may differ in humans. PMID- 14735495 TI - Rapid PKA-catalysed phosphorylation of boar sperm proteins induced by the capacitating agent bicarbonate. AB - In boar spermatozoa, the capacitating agent bicarbonate has been shown to induce rapid changes both in plasma membrane lipid architecture and in motility; in each case, a PKA-dependent pathway is involved. Early bicarbonate-induced changes in protein phosphorylation were probed using a commercial antibody against the phosphorylated form of the consensus substrate site for cyclic AMP-dependent protein kinase. The antibody detected relatively few bands in sperm extracts, of which only a small number showed incubation-dependent changes. While the quantitative response varied between boar ejaculates, in general terms bicarbonate induced phosphorylation increases in bands of 96, 64, and 59 kDa within 80 sec. The changes reached a maximum after about 160 sec, declined somewhat thereafter, and then increased again slowly as incubation progressed further (up to 21 min). The bicarbonate-induced increases were strongly dependent on the presence of BSA in the incubation medium. They were inhibited by H89 (PKA inhibitor) but not by GF (PKC inhibitor), and were enhanced by papaverine (phosphodiesterase inhibitor) and by calyculin (protein phosphatase inhibitor). The cyclic AMP analogue cBIMPS was able to mimic bicarbonate action though its effect was less dramatic. Stearated Ht31, a permeable inhibitor of PKA's binding to A-kinase anchoring protein, did not affect either the intensity or the specificity of the bicarbonate-induced phosphorylation changes, though it blocked motility entirely. Immunocytochemical studies revealed marked bicarbonate dependent phosphorylation changes in the post-acrosomal region of the head and in the neck, midpiece, and anterior regions of the tail. Fractionation of stimulated spermatozoa showed that all bands detectable with the antibody were bound to heads and to midpieces and associated large tail fragments; no bands were detected in either small tail or membrane fragments or in the cytoplasmic fraction. Differential extraction of the midpiece/large tail fraction revealed two protein bands with closely similar electrophoretic mobilities to the 96- and 59-kDa phosphorylated bands; MALDI-TOF analyses of these bands revealed both to be members of the Odf2 family. PMID- 14735496 TI - Human sperm aneuploidy after exposure to pesticides. AB - This study examined the effect of paternal environmental exposure to pesticides on the frequency of aneuploidy in human sperm. To determine if the chromosome number in germ cells was altered by paternal exposure, multicolor fluorescence in situ hybridization (FISH) analysis was utilized to measure aneuploidy frequencies in the sperm of 40 men (20 exposed, 20 controls). Samples were coded for "blind analysis" to eliminate scorer bias. Aneuploidy and diploidy frequencies were assessed for chromosomes 13, 21, X, and Y. A minimum of 10,000 sperm was scored per donor per chromosome probe with a total of 809,935 sperm scored. Hybridization efficiency was 99%. There were no significant differences in aneuploidy or diploidy frequencies between exposed and control groups, suggesting that the pesticides did not increase the risk of numerical chromosomal abnormalities in these men. PMID- 14735497 TI - Centrosomal function assessment in human sperm using heterologous ICSI with rabbit eggs: a new male factor infertility assay. AB - Sperm centrosomal function was assessed by immunocytochemical analysis after the injection of human sperm into mature rabbit eggs. Three hours after intracytoplasmic sperm injection (ICSI), an astral microtubule array from the base of the human sperm was observed in the rabbit eggs. This sperm aster expanded in the egg cytoplasm, concomitant with pronuclear formation, and a dense microtubule array was organized at the time of pronuclear centration. Using fertile donor sperm, the sperm aster formation rate at 3 hr after ICSI was 35.0 +/- 1.5%. Using sperm from infertile patients, the average aster formation rate was lower (25.4 +/- 14.8%, P<0.05). Among infertile cases, there was no correlation between sperm aster formation rates and conventional parameters of semen analysis. However, the sperm aster formation rate correlated with the embryonic cleavage rate following human in vitro fertilization (IVF). These data suggest that this assay reflects sperm function during embryonic development after sperm entry and that reproductive success during the first cell cycle requires a functional sperm centrosome. Furthermore, sperm centrosomal function cannot be predicted from conventional parameters of semen analysis. We propose that insufficient centrosomal function could be the cause of certain cases of idiopathic infertility. These assays may lead to the discovery of new types of infertility, which have previously been treated as "unexplained infertility," and may also lead to the treatment of infertility incurable even by ICSI. Consequently, an accurate and relevant assay to help assure couples of the success of fertilization is warranted, perhaps prior to ICSI therapy. PMID- 14735498 TI - MAP kinases regulate unfertilized egg apoptosis and fertilization suppresses death via Ca2+ signaling. AB - The default fate for eggs from many species is death by apoptosis and thus, successful fertilization depends upon suppression of the maternal death program. Little is known about the molecular triggers which activate this process or how the fertilization signal suppresses the default maternal apoptotic pathway. The MAP kinase (MAPK) family member, ERK, plays a universal and critical role in several stages of oocyte meiotic maturation, and fertilization results in ERK inactivation. In somatic cells, ERK and other MAPK family members, p38 and JNK, provide opposing signals to regulate apoptosis, however, it is not known whether MAPKs play a regulatory role in egg apoptosis, nor whether suppression of apoptosis by fertilization is mediated by MAPK activity. Here we demonstrate that MAPKs are involved in starfish egg apoptosis and we investigate the relationship between the fertilization induced signaling pathway and MAPK activation. ERK is active in post-meiotic eggs just until apoptosis onset and then p38, JNK and a third kinase are activated, and remain active through execution. Sequential activation of ERK and p38 is necessary for apoptosis, and newly synthesized proteins are required both upstream of ERK and downstream of p38 for activation of the full apoptotic program. Fertilization causes a dramatic rise in intracellular Ca2+, and we report that Ca2+ provides a necessary and sufficient pro-survival signal. The Ca2+ pathway following fertilization of both young and aged eggs causes ERK to be rapidly inactivated, but fertilization cannot rescue aged eggs from death, indicating that ERK inactivation is not sufficient to suppress apoptosis. PMID- 14735500 TI - Stabilization of p-block organoelement terminal hydroxides, thiols, and selenols requires newer synthetic strategies. AB - Metal hydroxides represent a very interesting and highly useful class of compounds that have been known to chemists for a very long time. While alkali and alkaline earth metal hydroxides (s-block) are commonplace chemicals in terms of their abundance and their use in a chemical laboratory as bases, the interest in Bronsted acidic molecular terminal hydroxides of p-block elements, such as aluminum and silicon, has been of recent origin, with respect to the variety of applications these compounds can offer both in materials science and catalysis. Moreover, these systems are environmentally friendly, relative to the metal halides, owing to their -OH functionality (resembling that of water). Design and conceptualization of the corresponding terminal thiols, selenols, and tellurols (M-SH, M-SeH, and M-TeH) offer even more challenging problems to synthetic inorganic chemists. This concept summarizes some of the recent strategies developed to stabilize these otherwise very unstable species. The successful preparation of a number of silicon trihydroxides a few years back resulted in the generation of several model compounds for metal-silicates. The recent synthesis of unusual aluminum compounds such as RAl(OH)(2), RAl(SH)(2), and RAl(SeH)(2) with terminal EH (E=O, Se, or Se) groups is likely to change the ways in which some of the well-known catalytic conversions are being carried out. The need for very flexible and innovative synthetic strategies to achieve these unusual compounds is emphasized in this concept. PMID- 14735501 TI - High oxidation state organometallic chemistry in aqueous media: new opportunities for catalysis and electrocatalysis. AB - Molybdenum has been used as a case study for the rational development of a new domain of organometallic chemistry, combining oxo-stabilized high oxidation state metals and aqueous media. The pH-dependent conversion of oxo, hydroxo, and aqua ligands allows the opening of free coordination sites, while the use of a redox active metal opens possible pathways for coupled proton and electron transfer to and from activated substrates. While our work has so far only concentrated on synthesis and analytical investigations, the development of catalytic and electrocatalytic processes in water or aqueous media is a promising application of this new research area. PMID- 14735502 TI - Sol-gel polycondensation of tetraethyl orthosilicate (TEOS) in sugar-based porphyrin organogels: inorganic conversion of a sugar-directed porphyrinic fiber library through sol-gel transcription processes. AB - Sugar-appended porphyrins (1 a-e) with monosaccharide groups at their periphery have been rationally designed for a new class of gelating reagents. A few of these compounds have the tendancy to form one-dimensional aggregates stable enough to show successful gelation ability for DMF-alcohol mixed solvents. The aggregation mode in the specific columnar super structures has been evaluated in detail by UV-visible spectrometry (UV/Vis), circular dichroism (CD), scanning electron microscopy (SEM), and transmission electron microscopy (TEM). All UV visible spectra of sugar-appended porphyrinic gels obtained from 1 a-c exhibit Soret band absorptions, which shift to lower wavelength and are significantly broadened. This phenomenon indicates that these porphyrin cores strongly interact with each other in an H-aggregate fashion, which drives the generation of a one dimensional porphyrin-stacking array. The CD spectra of the organogels from 1 a and 1 b, which are in anomers, exhibit an almost symmetric pattern, whereas the gel from 1 c gives a completely different pattern. This implies that the gel fibrils wind themselves in a right- or left-handed fashion; this reflects chirality in the specific molecular structure of the gelators. The results from SEM for the gel fibrils are in good agreement with the CD patterns. The gel fibrils in 1 a possess left-handed helicity, whereas those in 1 b wind themselves right-handedly. Macroscopic helical morphology reflects the microscopic structure well at a molecular level, which gives structural variety of the gel fibrils, which can be defined by the sugar library. Inorganic conversion of the organic helical fibrils by a sol-gel transcription process successfully gives the helical silica structures, which finely inherit the organic morphology. A striking observation is that a unimolecular porphyrin-stacking array is also transcribed into silica fibers when the optimized sol-gel reaction conditions are selected. A sugar-based organic-fiber library in porphyrinic gels thus provides a variety of inorganic materials through the sol-gel transcription process. PMID- 14735503 TI - Self-organized glycoclusters along DNA: effect of the spatial arrangement of galactoside residues on cooperative lectin recognition. AB - We describe herein the relationship between the spatial arrangement of self organized galactose clusters and lectin recognition. beta-Galactose-modified deoxyuridine phosphoramidite was synthesized and applied to solid-phase synthesis to provide 18-, 20-, and 22-mers of site-specifically galactosylated oligodeoxynucleotides (Gal-ODNs). These Gal-ODNs were self-organized through hybridization with the corresponding 18-, 20-, and 22-mers of half-sliding complementary ODNs (hsc-ODNs) to give periodic galactoside clusters. The self organization of ODNs was confirmed by size exclusion chromatography and gel electrophoresis. The binding of the Gal-clusters to the FITC-labeled RCA(120) lectin was analyzed by monitoring the change in fluorescence intensity. The assembly of 20-mer Gal-ODN with the 20-mer hsc-ODN was strongly and cooperatively recognized by the lectin. The 18-mer assembly was bound more weakly and less cooperatively, and the 22-mer assembly was minimally bound to the lectin. RCA(120) lectin recognized not only the density of galactoside residues, but also the spatial arrangement. The size of the Gal cluster was estimated from the association constant of Gal-ODN with hsc-ODN. The relationship between lectin recognition and Gal-cluster size is also discussed. PMID- 14735504 TI - Extension of conjugation leading to bathochromic or hypsochromic effects in OPV series. AB - Four OPV series 1-4 (a-d) with a terminal dialkylamino group as electron donor were prepared by Wittig-Horner reactions. To study the influence of the push-pull effect on the long-wavelength absorption, three of the four series contained terminal acceptor groups (CN, CHO, NO(2)). The length of the chromophores strongly affects the intramolecular charge transfer (ICT)-an effect which superimposes upon the extension of the conjugation. Increasing numbers n of repeat units cause an overall bathochromic shift for the purely donor-substituted series 1 a-4 a and the series 1 b-4 b with CN as weak acceptor. The two effects annihilate each other in the series 1 c-4 c with terminal CHO groups, so that the absorption maxima are almost independent of the length of the chromophore. A hypsochromic shift is observed for the series 1 d-4 d, which contains the strong acceptor group NO(2). This anomaly disappears on protonation of the dialkylamino group because the push-pull effect disappears in the ammonium salts. The results can be explained by semiempirical quantum mechanics (AM1, INDO/S). The HOMO-LUMO transition, which is mainly responsible for the ICT, becomes less important in the electron transitions S(0)-->S(1) when the distance between donor and acceptor is increased. The commonly used VB model, which contains an electroneutral and a zwitterionic resonance structure, is contrasted with a MO model with dipole segments at both ends of the OPV chains. The latter model turned out to be more appropriate-at least for donor-acceptor-substituted OPVs with n >/= 2. PMID- 14735505 TI - Enthalpy of solution of 1,4-naphthoquinone in CO2 + n-pentane in the critical region of the binary mixture: mechanism of solubility enhancement. AB - The enthalpy of solution (Delta(solv)H(m)) and solubility of 1,4-naphthoquinone in CO(2) + n-pentane were measured at 308.15 K in the critical region of the binary fluid. In order to study the effect of phase behavior of the mixed solvent on Delta(solv)H(m), the experiments were carried out in the supercritical (SC) and subcritical region of the binary solvent. The density of the mixed solvent in different conditions was determined. The isothermal compressibility (K(T)) of the mixed solvent, and the partial molar volume (V(n-pentane)) of n-pentane in the solution were calculated. It was demonstrated that the Delta(solv)H(m) was negative in all conditions. Delta(solv)H(m) is nearly independent of pressure or density in all the solvents in a high-density region, in which compressibility of the solvent is very small; this indicates that the intermolecular interaction between the solvent and the solute is similar to that for liquid solutions. It is very interesting that Delta(solv)H(m) in the mixed SC fluid differs from the Delta(solv)H(m) in mixed subcritical fluids. The absolute value of Delta(solv)H(m) in the mixed SC fluid is close to that in pure SC CO(2) in the high-density region, and is much lower than that in pure SC CO(2) in the low density region. In the mixed subcritical fluids, the Delta(solv)H(m) is also close to that in the pure CO(2) in the high-density region. However, at the same density, the absolute value of Delta(solv)H(m) in the binary subcritical fluid is larger than that in pure CO(2) in the high-compressible region of the mixed solvent. The main reason for this is that the degree of clustering in the SC solutions is small at the density in which the degree of clustering is large in the subcritical solutions. It can be concluded that solubility enhancement by n pentane in the mixed SC fluid is entropy driven. In contrast, the solubility enhancement by n-pentane in subcritical fluids is enthalpy driven. The intermolecular interaction in the SC solutions and subcritical solutions can be significantly different even if their densities are the same. PMID- 14735506 TI - Syntheses and reactivities of stable halosilylenoids, (Tsi)X2SiLi (Tsi=C(SiMe3)3, X=Br, Cl). AB - Halosilylenoids, stable at room temperature (Tsi)X(2)SiLi (Tsi=C(SiMe(3))(3), X=Br, Cl), were synthesized from the reaction of TsiSiX(3) with lithium naphthalenide. Bromosilylenoid reacted with tBuOH and MeI both at -78 degrees C and at room temperature to give (Tsi)HSiBr(2) and (Tsi)MeSiBr(2), respectively, in high yields; this clearly shows its nucleophilicity. In the reaction of bromosilylenoid with methanol, 2-propanol, and 2,3-dimethyl-1,3-butadiene, the corresponding products, (Tsi)HSi(OMe)(2), (Tsi)HSi(OiPr)Br, and bromo(Tsi)silacyclopent-3-ene, were obtained in high yields; this demonstrates its amphiphilic property, as if bromosilylene would be trapped. Chlorosilylenoid also exhibited both nucleophilic and amphiphilic properties. The (29)Si chemical shifts for (Tsi)Br(2)SiLi, (Tsi)Br(2)SiK, and (Tsi)Cl(2)SiLi were 106, 70, and 87 ppm, respectively. PMID- 14735507 TI - An inorganic double helix sheathing alkali metal cations: ANb2P2S12 (A=K, Rb, Cs), a series of thiophosphates close to the metal-nonmetal boundary- chalcogenide analogues of transition-metal phosphate bronzes? AB - The new quaternary niobium thiophosphates ANb(2)P(2)S(12) (A=K, Rb, Cs) have been prepared and characterized. The title compounds were synthesized by reacting Nb metal, A(2)S, P(2)S(5), and S at 600-700 degrees C in evacuated silica tubes. They crystallize as "stuffed" variants of the tetragonal TaPS(6) structure type in the tetragonal space group I$?bar 4$2d with eight formula units per unit cell and lattice constants a=15.923(2) and c=13.238(3) A for CsNb(2)P(2)S(12), a=15.887(3) and c=13.132(3) A for RbNb(2)P(2)S(12), and a=15.850(2) and c=13.119(3) A for KNb(2)P(2)S(12). Their structures are based on double helices formed from interpenetrating, noninteracting spiral chains of binuclear [Nb(2)S(12)] cluster units and [PS(4)] thiophosphate groups. The cavities and tunnels, which are formed by the helical chains, are filled with A(+) ions. Temperature-dependent conductivity studies reveal thermally activated electrical transport behavior. This result is consistent with the observation of a temperature-dependent contribution to the (31)P MAS-NMR shift, suggesting that the delocalized s-electron spin density increases with increasing temperature. These findings are supported by the results of tight-binding band structure calculations which reveal that the unusual electrical transport behavior of ANb(2)P(2)S(12) is a consequence of the structure symmetry. Therefore, CsNb(2)P(2)S(12) may be considered a chalcogenide analogue of metal phosphate bronzes. PMID- 14735508 TI - Chemical tailoring of porous silica xerogels: local structure by vibrational spectroscopy. AB - Monolithic porous silica xerogels were synthesized by the sol-gel process, and their local structure was analysed by vibrational spectroscopy. The silica alcogels were prepared by a two-step hydrolytic polycondensation of tetraethoxysilane (TEOS) in isopropanol, with a water/TEOS molar ratio of 4. The hydrolysis step was catalysed by hydrochloric acid (HCl), with different HCl/TEOS molar ratios (ranging from 0.0005 to 0.009), and the condensation step was catalysed by ammonia (NH(3)), with different NH(3)/HCl molar ratios (ranging from 0.7 to 1.7). After appropriate ageing, the alcogels were washed with isopropanol and subcritically dried at atmospheric pressure. The diffuse reflectance infrared Fourier transform (DRIFT) spectra were analysed in terms of the main siloxane rings that form the silica particles, taking into account the splitting of the nu(as)Sibond;Obond;Si mode into pairs of longitudinal and transverse optic components, by long-range Coulomb interactions. It was proven that the proportion of residual silanol groups (which correlates with hydrophilicity), and the fraction of siloxane 6-rings (which correlates with porosity) may be tailored by adequate catalytic conditions, mostly by the hydrolysis pH. This was explained in terms of the reactions' mechanisms taking place in the two-step sol-gel process followed. PMID- 14735509 TI - From D-glucose to biologically potent L-hexose derivatives: synthesis of alpha-L iduronidase fluorogenic detector and the disaccharide moieties of bleomycin A2 and heparan sulfate. AB - A novel and convenient route for the synthesis of biologically potent and rare L hexose derivatives from D-glucose is described. Conversion of diacetone-alpha-D glucose (14) into 1,2:3,5-di-O-isopropylidene-beta-L-idofuranose (19) was efficiently carried out in two steps. Orthogonal isopropylidene rearrangement of compound 19 led to 1,2:5,6-di-O-isopropylidene-beta-L-idofuranose (27), which underwent regioselective epimerization at the C3 position to give the L-talo- and 3-functionalized L-idofuranosyl derivatives. Hydrolysis of compound 19 under acidic conditions furnished 1,6-anhydro-beta-L-idopyranose (35) in excellent yield, which was successfully transformed into the corresponding L-allo, L-altro, L-gulo, and L-ido derivatives via regioselective benzylation, benzoylation, triflation and nucleophilic substitution as the key steps. Applications of these 1,6-anhydro-beta-L-hexopyranoses as valuable building blocks to the syntheses of 4-methylcoumarin-7-yl-alpha-L-iduronic acid and the disaccharide moieties of bleomycin A(2) as well as heparan sulfate are highlighted. PMID- 14735510 TI - Insertion of alkynes into the Pt-Si bond of silylplatinum complexes leading to the formation of 4-sila-3-platinacyclobutenes and 5-sila-2-platina-1,4 cyclohexadienes. AB - The reaction of dimethyl acetylenedicarboxylate (DMAD) with [Pt(SiHPh(2))(2)(PMe(3))(2)] produces cis-[Pt(CZ=CZ SiHPh(2))(SiHPh(2))(PMe(3))(2)] (cis-1, Z = COOMe) and [Pt(CZ=CZ SiPh(2))(PMe(3))(2)] (2) depending on the reaction conditions. cis-1 and 2 are equilibrated in solution at room temperature, and they are isolated by recrystallization of the mixtures. cis-1 is converted slowly in solution into trans-[Pt(CZ=CZ-SiHPh(2))(SiHPh(2))(PMe(3))(2)] (trans-1) via intermediate 2 followed by reaction with H(2)SiPh(2). DMAD also reacts with [Pt(SiHPh(2))(2)(dmpe)] (dmpe = 1,2-bis(dimethylphosphino)ethane) to afford [Pt(CZ=CZ-SiHPh(2))(SiHPh(2))(dmpe)] (3). Conversion of 3 into 4-sila-3 platinacyclobutene [Pt(CZ=CZ-SiPh(2))(dmpe)] (4) takes place, accompanied by formation of H(2)SiPh(2), to give an equilibrated mixture of the two complexes. Crystallographic and spectroscopic data of cis-1, trans-1, and 3 suggest the presence of an intramolecular interaction between the Si-H group of the 3-sila-1 propenyl ligand and Pt via an Si-H-Pt three-center-four-electron bond in the solid state and in solution. DMAD reacts with 2 to give 5-sila-2-platina-1,4 cyclohexadiene with pi-coordinated DMAD, [Pt(CZ=CZ-SiPh(2) CZ=CZ)(DMAD)(PMe(3))(2)] (5), which is also obtained from the reaction of excess DMAD with [Pt(SiHPh(2))(2)(PMe(3))(2)]. Unsymmetrical six-membered silaplatinacycles without pi-coordinated alkyne, [Pt(CZ=CZ-SiPh(2) CH=CX)(PMe(3))(2)] (6: X = COOMe; 7: X = Ph), are prepared analogously from the respective reactions of phenyl acetylene and of methyl acetylene carboxylate with 2. Methyl 2-butynolate reacts with 2 at 50 degrees C to form a mixture of the regioisomers [Pt(CZ=CZ-SiPh(2)-CMe=CZ)(PMe(3))(2)] (8) and [Pt(CZ=CZ-SiPh(2) CZ=CMe)(PMe(3))(2)] (9). PMID- 14735512 TI - Synthesis of scroll-type composite microtubes of Mo2C/MoCO by controlled pyrolysis of Mo(CO)6. AB - Composite microtubes of Mo(2)C/MoCO have been synthesized for the first time under well-controlled conditions by thermal decomposition of Mo(CO)(6) at about 600 degrees C. Here, thermal stability and phase transition of the products, as well as the influence of reaction temperature and argon flow rate, have been carefully investigated. All samples were characterized by X-ray powder diffraction (XRD), scanning electron microscopy (SEM), and X-ray photoelectron spectroscopy (XPS). The reaction model and rolling mechanism were proposed on the basis of the experimental facts. PMID- 14735511 TI - New chiral ruthenium(II) catalysts containing 2,6-bis(4'-(R)-phenyloxazolin-2' yl)pyridine (Ph-pybox) ligands for highly enantioselective transfer hydrogenation of ketones. AB - Treatment of complex trans-[RuCl(2)(eta(2)-C(2)H(4))[kappa(3)-N,N,N-(R,R)-Ph pybox]] [(R,R)-Ph-pybox = 2,6-bis[4'-(R)-phenyloxazolin-2'-yl]pyridine] with phosphines or phosphites in dichloromethane at 50 degrees C leads to the formation of novel ruthenium(II)-pybox complexes trans-[RuCl(2)(L)[kappa(3)-N,N,N (R,R)-Ph-pybox]] [L = PPh(3) (1 a), PPh(2)Me (2 a), PPh(2)(C(3)H(5)) (3 a), PPh(2)(C(4)H(7)) (4 a), PMe(3) (5 a), PiPr(3) (6 a), P(OMe)(3) (7 a) and P(OPh)(3) (8 a)]. Likewise, reaction of trans-[RuCl(2)(eta(2)-C(2)H(4))[kappa(3) N,N,N-(R,R)-Ph-pybox]] with PPh(3) or PiPr(3) in refluxing methanol leads to the complexes cis-[RuCl(2)(L)(kappa(3)-N,N,N-(R,R)-Ph-pybox] [L = PPh(3) (1 b), PiPr(3) (6 b)]. No trans-cis isomerisation of complexes 1 a-8 a has been observed. Complexes 1 a-8 a, 1 b, 6 b together with the analogous trans [RuCl(2)[P(OMe)(3)][kappa(3)-N,N,N-(S,S)-iPr-pybox]] (10 a) and the previously reported trans- and cis-[RuCl(2)(PPh(3))[kappa(3)-N,N,N-(S,S)-iPr-pybox]] (9 a and 9 b, respectively) are active catalysts for the transfer hydrogenation of acetophenone in 2-propanol in the presence of NaOH (ketone/cat/NaOH 500:1:6). cis Ph-pybox derivatives are the most active catalysts. In particular, cis complexes 1 b and 6 b led to almost quantitative conversions in less than 5 min with a high enantioselectivity (up to 95 %). A variety of aromatic ketones have also been reduced to the corresponding secondary alcohols with very high TOF and ee up to 94 %. The overall catalytic performance seems to be a subtle combination of the steric and/or electronic properties both the phosphines and the ketones. A high TOF (27 300 h(-1)) and excellent ee (94 %) have been found for the reduction of 3 bromoacetophenone with catalyst 6 b. Reductions of alkyl ketones also proceed with high and rapid conversions but low enantioselectivities are achieved. PMID- 14735513 TI - Photohydroxylation of 1,4-benzoquinone in aqueous solution revisited. AB - In water, photolysis of 1,4-benzoquinone, Q gives rise to equal amounts of 2 hydroxy-1,4-benzoquinone HOQ and hydroquinone QH(2) which are formed with a quantum yield of Phi=0.42, independent of pH and Q concentration. By contrast, the rate of decay of the triplet (lambda(max)=282 and approximately 410 nm) which is the precursor of these products increases nonlinearly (k=(2-->3.8) x 10(6) s( 1)) with increasing Q concentration ((0.2-->10) mM). The free-radical yield detected by laser flash photolysis after the decay of the triplet also increases with increasing Q concentration but follows a different functional form. These observations are explained by a rapid equilibrium of a monomeric triplet Q* and an exciplex Q(2)* (K=5500+/-1000 M(-1)). While Q* adds water and subsequent enolizes into 1,2,4-trihydroxybenzene Ph(OH)(3), Q(2)* decays by electron transfer and water addition yielding benzosemiquinone (.)QH and (.)OH adduct radicals (.)QOH. The latter enolizes to the 2-hydroxy-1,4-semiquinone radical (.)Q(OH)H within the time scale of the triplet decay and is subsequently rapidly (microsecond time scale) oxidized by Q to HOQ with the concomitant formation of (.)QH. On the post-millisecond time scale, that is, when (.)QH has decayed, Ph(OH)(3) is oxidized by Q yielding HOQ and QH(2) as followed by laser flash photolysis with diode array detection. The rate of this pH- and Q concentration dependent reaction was independently determined by stopped-flow. This shows that there are two pathways to photohydroxylation; a free-radical pathway at high and a non-radical one at low Q concentration. In agreement with this, the yield of Ph(OH)(3) is most pronounced at low Q concentration. In the presence of phosphate buffer, Q* reacts with H(2)PO(4) (-) giving rise to an adduct which is subsequently oxidized by Q to 2-phosphato-1,4-benzoquinone QP. The current view that (.)OH is an intermediate in the photohydroxylation of Q has been overturned. This view had been based on the observation of the (.)OH adduct of DMPO when Q is photolyzed in the presence of this spin trap. It is now shown that Q*/Q(2)* oxidizes DMPO (k approximately 1 x 10(8) M(-1) s(-1)) to its radical cation which subsequently reacts with water. Q*/Q(2)* react with alcohols by H abstraction (rates in units of M(-1) s(-1)): methanol (4.2 x 10(7)), ethanol (6.7 x 10(7)), 2 propanol (13 x 10(7)) and tertiary butyl alcohol ( approximately 0.2 x 10(7)). DMSO (2.7 x 10(9)) and O(2) ( approximately 2 x 10(9)) act as physical quenchers. PMID- 14735514 TI - Stereocontrolled synthesis of 8,11-dideoxytetrodotoxin, an unnatural analogue of puffer fish toxin. AB - 8,11-Dideoxytetrodotoxin, an unnatural tetrodotoxin analogue, was synthesized in a highly stereoselective manner from a common intermediate from our synthetic studies on tetrodotoxin. The key features in the synthesis were as follows: neighboring group participation of a trichloroacetamide to allow regioselective and stereoselective hydroxylation, protection of a delta-hydroxylactone as an ortho ester, and guanidine installation through the use of Boc-protected isothiourea. Global deprotection of the fully protected intermediate under acidic conditions gave 8,11-dideoxytetrodotoxin, which exhibited very weak biological activities. PMID- 14735515 TI - Solid-phase synthesis of 89 polyamine-based cationic lipids for DNA delivery to mammalian cells. AB - The ability of non-viral gene delivery systems to overcome extracellular and intracellular barriers is a critical issue for future clinical applications of gene therapy. In recent years much effort has been focused on the development of a variety of DNA carriers, and cationic liposomes have become the most common non viral gene delivery system. Solid-phase synthesis was used to produce three libraries of polyamine-based cationic lipids with diverse hydrophobic tails. These were characterised, and structure-activity relationships were determined for DNA binding and transfection ability of these compounds when formulated as cationic liposomes. Two of the cationic lipids produced high-efficiency transfection of human cells. Surprisingly, these two compounds were from the library with two headgroups and one aliphatic tail, a compound class regarded as detergent-like and little investigated for transfection. These cationic lipids are promising reagents for gene delivery and illustrate the potential of solid phase synthesis methods for lipoplex discovery. PMID- 14735516 TI - Effects of metal ions on photoinduced electron transfer in zinc porphyrin naphthalenediimide linked systems. AB - Zinc porphyrin-naphthalenediimide (ZnP-NIm) dyads and zinc porphyrin pyromellitdiimide-naphthalenediimide (ZnP-Im-NIm) triad have been employed to examine the effects of metal ions on photoinduced charge-separation (CS) and charge-recombination (CR) processes in the presence of metal ions (scandium triflate (Sc(OTf)(3)) or lutetium triflate (Lu(OTf)(3)), both of which can bind with the radical anion of NIm). Formation of the charge-separated states in the absence and in the presence of Sc(3+) was confirmed by the appearance of absorption bands due to ZnP(.) (+) and NIm(.) (-) in the absence of metal ions and of those due to ZnP(.) (+) and the NIm(.) (-)/Sc(3+) complex in the presence of Sc(3+) in the time-resolved transient absorption spectra of dyads and triad. The lifetimes of the charge-separated states in the presence of 1.0 x 10(-3) M Sc(3+) (14 micros for ZnP-NIm, 8.3 micros for ZnP-Im-NIm) are more than ten times longer than those in the absence of metal ions (1.3 micros for ZnP-NIm, 0.33 micros for ZnP-Im-NIm). In contrast, the rate constants of the CS step determined by the fluorescence lifetime measurements are the same, irrespective of the presence or absence of metal ions. This indicates that photoinduced electron transfer from (1)ZnP(*) to NIm in the presence of Sc(3+) occurs without involvement of the metal ion to produce ZnP(.) (+)-NIm(.) (-), followed by complexation with Sc(3+) to afford the ZnP(.) (+)-NIm(.) (-)/Sc(3+) complex. The one-electron reduction potential (E(red)) of the NIm moiety in the presence of a metal ion is shifted in a positive direction with increasing metal ion concentration, obeying the Nernst equation, whereas the one-electron oxidation potential of the ZnP moiety remains the same. The driving force dependence of the observed rate constants (k(ET)) of CS and CR processes in the absence and in the presence of metal ions is well evaluated in terms of the Marcus theory of electron transfer. In the presence of metal ions, the driving force of the CS process is the same as that in the absence of metal ions, whereas the driving force of the CR process decreases with increasing metal ion concentration. The reorganization energy of the CR process also decreases with increasing metal ion concentration, when the CR rate constant becomes independent of the metal ion concentration. PMID- 14735517 TI - Evidence that protons can be the active catalysts in Lewis acid mediated hetero Michael addition reactions. AB - The mechanism of Lewis acid catalysed hetero-Michael addition reactions of weakly basic nucleophiles to alpha,beta-unsaturated ketones was investigated. Protons, rather than metal ions, were identified as the active catalysts. Other mechanisms have been ruled out by analyses of side products and of stoichiometric enone catalyst mixtures and by the use of radical inhibitors. No evidence for the involvement of pi-olefin-metal complexes or for carbonyl-metal-ion interactions was obtained. The reactions did not proceed in the presence of the non coordinating base 2,6-di-tert-butylpyridine. An excellent correlation of catalytic activities with cation hydrolysis constants was obtained. Different reactivities of mono- and dicarbonyl substrates have been rationalised. A (1)H NMR probe for the assessment of proton generation was established and Lewis acids have been classified according to their propensity to hydrolyse in organic solvents. Bronsted acid-catalysed conjugate addition reactions of nitrogen, oxygen, sulfur and carbon nucleophiles are developed and implications for asymmetric Lewis acid catalysis are discussed. PMID- 14735518 TI - Palladium-catalyzed cross-coupling reactions of amines with alkenyl bromides: a new method for the synthesis of enamines and imines. AB - The palladium-catalyzed cross-coupling reaction of alkenyl bromides with secondary and primary amines gives rise to enamines and imines, respectively. This new transformation expands the applicability of palladium-catalyzed C-N bond forming reactions (the Buchwald-Hartwig amination), which have mostly been applied to aryl halides. After screening of different ligands, bases, and solvents, the catalytic combination [Pd(2)(dba)(3)]/BINAP in the presence of NaOtBu in toluene gave the best results in the cross-coupling of secondary amines with 1-bromostyrene (dba=dibenzylideneacetone, BINAP=2,2'-bis(diphenylphosphino) 1,1'-binaphthyl). The corresponding enamines are obtained cleanly and in nearly quantitative yields. However, steric hindrance seems to be a limitation of the reaction, as amines carrying large substituents are not well converted. The same methodology can be applied to the coupling of secondary amines with 2 bromostyrene. Moreover, the reaction with substituted 2-bromopropenes allows regioselective synthesis of isomerizable terminal enamines without isomerization of the double bond. The best catalytic conditions for the cross-coupling of 1 bromostyrene with primary amines include again the use of the Pd(0)/BINAP/NaOtBu system. The reaction gives rise to the expected imines in very short times and with low catalyst loadings. A set of structurally diverse imines can be prepared by this methodology through variations in the structure of both coupling partners. However, 2-bromostyrene failed to give good results in this coupling reaction, probably due to product inhibition of the catalytic cycle. Competition experiments of vinyl versus aryl amination reveal that the reaction occurs preferentially on vinyl bromides. PMID- 14735519 TI - First structural characterization of binary AsIII and SbIII azides. AB - The highly explosive molecules As(N(3))(3) and Sb(N(3))(3) were obtained in pure form by the reactions of the corresponding fluorides with (CH(3))(3)SiN(3) in SO(2) and purification by sublimation. The crystal structures and (14)N NMR, infrared, and Raman spectra were determined, and the results compared to ab initio second-order perturbation theory calculations. Whereas Sb(N(3))(3) possesses a propeller-shaped, pyramidal structure with perfect C(3) symmetry, the As(N(3))(3) molecule is significantly distorted from C(3) symmetry due to crystal packing effects. PMID- 14735520 TI - Benzofuran trimers for organic electroluminescence. AB - Four linear benzofuran trimers have been prepared by a two-stage synthetic procedure. They were tested as materials for organic electroluminescence (OEL). Precursor phenylene ethynylene oligomers were formed in the first stage, then after removal of the phenolic hydroxyl protecting groups, a base was used to promote the cyclization of ortho-hydroxy phenylene ethynylenes to benzofurans. Both acetate esters and tert-butyl carbonates were employed as protecting groups. tert-Butyl and n-hexyl substituents on the benzofurans were used to modulate solubility, aggregation, and film-forming properties; two tert-butyl groups prevented aggregation in the solid state, thus maintaining emission in the blue region of the visible spectrum. The OEL characteristics of the tert-butyl substituted benzofuran trimer were explored, and blue emission was observed. The two-stage synthetic procedure employed for the preparation of these benzofuran trimers may be applied to a wide variety of benzofuran oligomer and polymer targets. PMID- 14735521 TI - Dendronized perylenetetracarboxdiimides with peripheral triphenylamines for intramolecular energy and electron transfer. AB - Novel perylene-3,4,9,10-tetracarboxdiimides (PDI) dyes functionalized with polyphenylene dendrimers attached at the bay region are reported. Derivatives of PDI bearing polyphenylene dendrimers up to the second generation, substituted with an increasing number of triphenylamine (TPA) moieties at the periphery, as well as a related nondendronized model compound were prepared. Intramolecular energy transfer was demonstrated by the observation of PDI emission on excitation of the triphenylamines, and electron transfer was detected by comparing photoluminescence quenching in solvents of different polarity. PMID- 14735522 TI - Interactions between organic molecules and water: rotational spectrum of the 1:1 oxetane-water complex. AB - The 1:1 molecular complex between oxetane and water has been investigated by using free-jet millimeter-wave spectroscopy. The rotational spectra of five isotopomers (with H(2)O, D(2)O, DOH, HOD and H(2) (18)O) have been assigned. Partial r(0) and r(s) structures of the complex have been derived. The water moiety lies in the plane of symmetry of oxetane, with the "free" hydrogen E with respect to the ring. The oxetane ring appears to be slightly nonplanar, with the C(beta) carbon tilted on the opposite side of the water unity. The three atoms involved in the hydrogen bond adopt a linear arrangement with an O(ring).H distance of about 1.86 A, and the angle between the COC bisector and the O(ring).H bond being congruent with 106 degrees. Additionally, quantum-chemical calculations for the complex were performed and were found to be in agreement with the experimental results. PMID- 14735523 TI - Enantioselective synthesis of non-natural aromatic alpha-amino acids. AB - We present two complementary methods for the stereoselective synthesis of non natural alpha-amino acids with aromatic or heteroaromatic side chains. One approach is based on the chemical transformation of methionine, whereas the other applies the stereoselective Myers alkylation of glycine. The resulting product types differ in the linker length between glycine and the aromatic substituent. Since methionine and pseudoephedrine are available in both absolute configurations, R- or S-configured enantiopure amino acids with either C(2) or C(3) linkers can be obtained on gram scales. In each case the key step of the synthesis is hydroboration of the unsaturated building blocks 9 and 17, followed by palladium-catalyzed Suzuki cross-coupling with aryl halides. Attention must in certain cases be paid to the stereochemical integrity when basic Suzuki conditions are applied. Our initial difficulties are reported as well as the final "racemization-proof" procedures. The protecting groups chosen for the alpha amino acids should be compatible with solid-phase peptide synthesis. This was confirmed by the successful synthesis of a series of tripeptides. PMID- 14735524 TI - Uniform micro-sized alpha- and beta-Si3N4 thin ribbons grown by a high temperature thermal-decomposition/nitridation route. AB - Uniform micro-sized alpha- and beta-Si(3)N(4) thin ribbons have been achieved by a high-temperature thermal-decomposition/nitridation route. As-grown ribbons were characterized by using powder X-ray diffraction, scanning electron microscopy, transmission electron microscopy, energy-dispersive X-ray spectroscopy, electron energy loss spectroscopy, and cathodoluminescence. These alpha- and beta Si(3)N(4) ribbons are structurally uniform micro-sized single crystals, and have a width of approximately 2-3 microns, a thickness of approximately 20-60 nm, and a length, that ranges from several hundreds of microns to the order of millimeters. A room-temperature cathodoluminescence spectrum recorded from these ribbons shows one intensive blue emission peak at approximately 433 nm. The growth for the new ribbon form of this material is believed to be dominated by a vapor-solid process. PMID- 14735528 TI - Do heavy nuclei see light at the end of the tunnel? PMID- 14735529 TI - Stable, self-assembling, equilibrating catalysts for green chemistry. PMID- 14735530 TI - Chemistry in microstructured reactors. AB - The application of microstructured reactors in the chemical process industry has gained significant importance in recent years. Companies that offer not only microstructured reactors, but also entire chemical process plants and services relating to them, are already in existence. In addition, many institutes and universities are active within this field, and process-engineering-oriented reviews and a specialized book are available. Microstructured systems can be applied with particular success in the investigation of highly exothermic and fast reactions. Often the presence of temperature-induced side reactions can be significantly reduced through isothermal operations. Although microstructured reaction techniques have been shown to optimize many synthetic procedures, they have not yet received the attention they deserve in organic chemistry. For this reason, this Review aims to address this by providing an overview of the chemistry in microstructured reactors, grouped into liquid-phase, gas-phase, and gas-liquid reactions. PMID- 14735531 TI - Metal-mediated cross-linking in the generation of a marine-mussel adhesive. PMID- 14735532 TI - Identification of potent Ras signaling inhibitors by pathway-selective phenotype based screening. PMID- 14735533 TI - Sulindac-derived Ras pathway inhibitors target the Ras-Raf interaction and downstream effectors in the Ras pathway. PMID- 14735534 TI - Spontaneous assembly of a monolayer of charged gold nanocrystals at the water/oil interface. PMID- 14735535 TI - X-ray absorption spectroscopic and electrochemical studies of tris(catecholato(2 ))chromate(V/IV/III) complexes. PMID- 14735536 TI - Helical superstructure of conductive polymers as created by electrochemical polymerization by using synthetic lipid assemblies as a template. PMID- 14735538 TI - One-dimensional magnetism in anhydrous iron and cobalt ternary oxalates with rare trigonal-prismatic metal coordination environment. PMID- 14735537 TI - Pressure-induced hydration effects in the zeolite laumontite. PMID- 14735539 TI - Heterobimetallic C3 complexes through silylpropargylidyne desilylation. PMID- 14735540 TI - An efficient synthesis of bicyclic amidines by intramolecular cyclization. PMID- 14735541 TI - Polymer-supported ionic liquids: imidazolium salts as catalysts for nucleophilic substitution reactions including fluorinations. PMID- 14735542 TI - Facile oxidative rearrangement of dispiro-porphodimethenes to nonplanar and sheetlike porphyrins with intense absorptions in the near-IR region. PMID- 14735543 TI - Stereoselective construction of acyclic carbon chains by a one-pot coupling process based on alkenyloxazoline-titanium complexes. PMID- 14735544 TI - Peptidyl thiophenols as substrates for nonribosomal peptide cyclases. PMID- 14735546 TI - Stable cubic phosphorus-containing radicals. PMID- 14735545 TI - An integrated approach to a portable and low-cost immunoassay for resource-poor settings. PMID- 14735547 TI - Two helical conformations from a single foldamer backbone: "split personality" in short alpha/beta-peptides. PMID- 14735548 TI - Surprisingly stable helical conformations in alpha/beta-peptides by incorporation of cis-beta-aminocyclopropane carboxylic acids. PMID- 14735549 TI - Titanium(IV)-catalyzed dynamic kinetic asymmetric transformation of alcohols, silyl ethers, and acetals under carbon allylation. PMID- 14735550 TI - Structural changes in nanoparticle catalysts as monitored by their magnetic properties. PMID- 14735551 TI - Facile route to ultraflat SAM-protected gold surfaces by "amphiphile splitting". PMID- 14735553 TI - Pathophysiology of acute graft-versus-host disease. AB - Graft-versus-host disease (GVHD) has been the primary limitation to the wider application of allogeneic bone marrow transplantation (BMT). The pathophysiology of acute GVHD is complex and can be conceptualized to be a three-step process based on murine studies. In step 1, the conditioning regimen leads to the damage and activation of host tissues and induces the secretion of inflammatory cytokines. As a consequence, the expression of MHC antigens and adhesion molecules is increased enhancing the recognition of host alloantigens by donor T cells. Donor T-cell activation in step 2 is characterized by donor T cell interaction with host APCs and subsequent proliferation, differentiation and secretion of cytokines. Cytokines such as IL-2 and IFN-gamma enhance T-cell expansion, induce cytotoxic T cells (CTL) and natural killer (NK) cell responses and prime additional mononuclear phagocytes to produce TNF-alpha and IL-1. These inflammatory cytokines in turn stimulate production of inflammatory chemokines, thus recruiting effector cells into target organs. In step 3, effector functions of mononuclear phagocytes are triggered via a secondary signal provided by lipopolysaccharide (LPS) that leaks through the intestinal mucosa damaged during step 1. This mechanism may result in the amplification of local tissue injury and further promotion of an inflammatory response, which, together with the CTL and NK components, leads to target tissue destruction in the transplant host. The following review discusses the three-step process of the pathophysiology of experimental acute GVHD. PMID- 14735554 TI - Low dose 2-CdA schedule activity in splenic marginal zone lymphomas. AB - Splenic Marginal Zone Lymphoma (SMZL) is a rare clinicopathological entity among marginal zone lymphomas. SMZL is an indolent lymphoma usually treated by splenectomy. A subset of patients is characterized by a more aggressive clinical course and poor prognosis. Treatment of these cases and second-line therapy for relapsed patients have not been yet identified. We report 10 cases treated with cladribrine (5 mg/m(2)/week) for six courses. Six patients (60%) achieved partial response, two patients (20%) achieved a complete response and the two remaining patients did not respond and died as a result of progression of the disease. The treatment was well tolerated. A total of 60% of the patients had an overall survival rate of 48 months and 24 months progression-free-survival was achieved by 37% with a median time of progression-free-survival of 17 months. Interestingly, in addition to a relevant percentage of hematological remission, some patients also experienced a molecular remission. We conclude that this treatment is safe and well tolerated and is able to induce a substantial number of responses. Our results suggest that this schedule is well tolerated and could be an useful alternative to splenectomy. PMID- 14735556 TI - Cellular phone use does not acutely affect blood pressure or heart rate of humans. AB - A recent study raised concern about increase of resting blood pressure after a 35 min exposure to the radiofrequency (RF) field emitted by a 900 MHz cellular phone. In this randomized, double blind, placebo controlled crossover trial, 32 healthy subjects were submitted to 900 MHz (2 W), 1800 MHz (1 W) cellular phone exposure, and to sham exposure in separate sessions. Arterial blood pressure (arm cuff method) and heart rate were measured during and after the 35 min RF and sham exposure sessions. We evaluated cardiovascular responses in terms of blood pressure and heart rate during controlled breathing, spontaneous breathing, head up tilt table test, Valsalva manoeuvre and deep breathing test. Arterial blood pressure and heart rate did not change significantly during or after the 35 min RF exposures at 900 MHz or 1800 MHz, compared to sham exposure. The results of this study indicate that exposure to a cellular phone, using 900 MHz or 1800 MHz with maximal allowed antenna powers, does not acutely change arterial blood pressure and heart rate. PMID- 14735555 TI - Epoetin alfa corrects anemia and improves quality of life in patients with hematologic malignancies receiving non-platinum chemotherapy. AB - Anemia, a commonly occurring morbidity in patients with cancer, often leads to diminished quality of life (QOL). Numerous clinical trials have shown that epoetin alfa treatment improves hematologic and QOL variables in cancer patients. The clinical trial analysis reported here was performed to assess response to epoetin alfa in patients with hematologic malignancies. Cancer patients with anemia undergoing non-platinum-based chemotherapy who were enrolled in a multinational, randomized (2:1), double-blind, placebo-controlled trial were prospectively stratified by tumor type (hematologic, solid). Efficacy endpoints included proportion of patients transfused after day 28; change in hemoglobin (Hb) level from baseline to last assessment; proportion of treatment responders (increase in Hb > or =2 g/dl unrelated to transfusion) and correctors (patients whose Hb levels reached > or =12 g/dl during the study); and QOL. The protocol was amended before unblinding to prospectively collect and assess survival data 12 months after the last patient completed the study, and survival for the full study cohort was estimated using Kaplan-Meier techniques. Efficacy analyses of hematologic and QOL variables, as well as Kaplan-Meier estimates of survival, were performed post hoc for the hematologic tumor stratum. Among patients with hematologic malignancies, the mean increase in Hb levels was greater with epoetin alfa than with placebo treatment (2.2 vs. 0.3 g/dl). Transfusion requirements were lower in patients who received epoetin alfa versus placebo (25.2 vs. 43.1%), and the proportion of responders and correctors was higher with epoetin alfa than with placebo (75.2 vs. 16.7% and 72.6 vs. 14.8%, respectively). Patients who received epoetin alfa had improved QOL while patients who received placebo had decreased QOL. These results are similar to those seen in the full study cohort, where differences between epoetin alfa and placebo were significant (P<0.05) for all five primary cancer- and anemia-specific QOL domains evaluated. Although the study was not powered for survival, Kaplan-Meier estimates showed a trend in overall survival favoring epoetin alfa in both the full study cohort and the hematologic subgroup. Epoetin alfa treatment was well tolerated. Epoetin alfa therapy increased Hb levels, reduced transfusion requirements, and improved QOL in patients with anemia undergoing non-platinum chemotherapy for hematologic malignancies. PMID- 14735557 TI - Apparent biological effect of strong magnetic field on mosquito egg hatching. AB - Apparent biological effects of strong magnetic fields were observed in the hatching behavior of fresh mosquito eggs in the center of 9.4 and 14.1 T magnets. In the first experiment performed at 20 +/- 1 degrees C, the hatching was delayed 32 h by a 9.4 T magnetic field and 71 h by a 14.1 T magnetic field. In the second experiment performed at 22 +/- 1 degrees C, the hatching was delayed 14 h by a 9.4 T magnetic field and 27 h by a 14.1 T magnetic field. In the magnetic field range of this study, the hatching delay increases nonlinearly with the intensity of the magnetic field. The experimental results also suggest that the biological effects of magnetic fields could be reversible or partially reversible to some extent. PMID- 14735558 TI - Geomagnetic field effect on cardiovascular regulation. AB - The goal of the present research was try to explain the physiological mechanism for the influence of the geomagnetic field (GMF) disturbance, reflected by the indices of the geomagnetic activity (K, K(p), A(k), and A(p) indices), on cardiovascular regulation. One hundred forty three experimental runs (one daily) comprising 50 min hemodynamic monitoring sequences were carried out in rabbits sedated by pentobarbital infusion (5 mg/kg/h). We examined the arterial baroreflex effects on the short term blood pressure and heart rate (HR) variabilities reflected by the standard deviation (SD) of the average values of the mean femoral arterial blood pressure (MAP) and the HR. Baroreflex sensitivity (BRS) was estimated from blood pressure/HR response to intravenous (i.v.) bolus injections of vasoconstrictor (phenylephrine) and vasodilator (nitroprusside) drugs. We found a significant negative correlation of increasing GMF disturbance (K(p)) with BRS (P = 0.008), HR SD (P =0.022), and MAP SD (P = 0.002) signifying the involvement of the arterial baroreflex mechanism. The abrupt change in geomagnetic disturbance from low (K = 0) to high (K = 4-5) values was associated with a significant increase in MAP (83 +/- 5 vs. 99 +/- 5 mm Hg, P = 0.045) and myocardial oxygen consumption, measured by MAP and HR product (24100 +/- 1800 vs. 31000 +/- 2500 mm Hg. bpm, P = 0.034), comprising an additional cardiovascular risk. Most likely, GMF affects brainstem and higher neural cardiovascular regulatory centers modulating blood pressure and HR variabilities associated with the arterial baroreflex. PMID- 14735559 TI - Importance of addressing National Electrical Code violations that result in unusual exposure to 60 Hz magnetic fields. AB - We evaluated wiring in multifamily developments containing National Electrical Code(R) (NEC(R)) violations as a source of unusual exposure to 60 Hz magnetic fields. Two methods were used in this evaluation: measurement and modeling. We measured the building wiring as a source of magnetic fields in six multifamily developments in Michigan. In this small sample, building wiring proved to be an important source of exposure in four of the six cases. In all four cases with exposure from building wiring, one or more NEC violations were involved. To supplement our measurement efforts, we used computer modeling to compare magnetic field exposure due to building wiring with magnetic field exposure from external power lines. Our calculations showed that where the building wiring has a NEC violation leading to net current loops, the exposure due to wiring is likely to be more important than that from external power lines. Our results support the results obtained in a recent study of the exposure of Californian K-12 students to magnetic fields, where building wiring with one or more NEC violation was found to be the single most important exposure source. If 60 Hz magnetic fields are important to avoid, then improved enforcement of the NEC, as required by law, is perhaps the single most important mitigation policy to adopt. Bioelectromagnetics 25:102-106, 2004. PMID- 14735560 TI - Corona discharge influences ozone concentrations near rats. AB - Ozone can be produced by corona discharge either in dry air or when one electrode is submerged in water. Since ozone is toxic, we examined whether ozone production by corona near laboratory animals could reach levels of concern. Male rats were exposed to a corona discharge and the concentration of ozone produced was measured. The resulting concentration of ozone ranged from ambient levels to 250 ppb when animals were located 1 cm from a 10 kV source. Similar ozone concentrations were observed when a grounded water source was present. Possible explanations for, as well as concerns regarding, ozone production under these conditions are discussed. PMID- 14735561 TI - Teratological studies of prenatal exposure of mice to a 20 kHz sawtooth magnetic field. AB - In order to evaluate the importance of gestational age in possible effects due to exposure to a 20 kHz sawtooth magnetic field, pregnant ICR mice at gestational 2.5-15.5 days post-coitus, which is the most sensitive stage for the induction of major congenital malformations, were exposed in a carrousel irradiator. The mice were exposed to a 20 kHz intermediate frequency (IF) sawtooth magnetic field had a 6.5 microT peak intensity for 8 h/day. The animals were sacrificed on the 18th day of gestation; and the fetuses were examined for mortality, growth retardation, changes in head size, and other morphological abnormalities. From the above conditions, it is concluded that the exposure to a 20 kHz sawtooth magnetic field with 6.5 microT peak intensity does not inflict any adverse effect on fetuses of pregnant mice. PMID- 14735562 TI - Low electromagnetic field (50 Hz) induces differentiation on primary human oral keratinocytes (HOK). AB - This work concerns the effect of low frequency electromagnetic fields (ELF) on biochemical properties of human oral keratinocytes (HOK). Cells exposed to a 2 mT, 50 Hz, magnetic field, showed by scanning electron microscopy (SEM) modification in shape and morphology; these modifications were also associated with different actin distribution, revealed by phalloidin fluorescence analysis. Moreover, exposed cells had a smaller clonogenic capacity, and decreased cellular growth. Indirect immunofluorescence with fluorescent antibodies against involucrin and beta-catenin, both differentiation and adhesion markers, revealed an increase in involucrin and beta-catenin expression. The advance in differentiation was confirmed by a decrease of expression of epidermal growth factor (EGF) receptor in exposed cells, supporting the idea that exposure to electromagnetic field carries keratinocytes to higher differentiation level. These observations support the hypothesis that 50 Hz electromagnetic fields may modify cell morphology and interfere in differentiation and cellular adhesion of normal keratinocytes. PMID- 14735563 TI - Apoptosis induced by ultraviolet radiation is enhanced by amplitude modulated radiofrequency radiation in mutant yeast cells. AB - The aim of this study was to investigate whether radiofrequency (RF) electromagnetic field (EMF) exposure affects cell death processes of yeast cells. Saccharomyces cerevisiae yeast cells of the strains KFy417 (wild-type) and KFy437 (cdc48-mutant) were exposed to 900 or 872 MHz RF fields, with or without exposure to ultraviolet (UV) radiation, and incubated simultaneously with elevated temperature (+37 degrees C) to induce apoptosis in the cdc48-mutated strain. The RF exposure was carried out in a special waveguide exposure chamber where the temperature of the cell cultures can be precisely controlled. Apoptosis was analyzed using the annexin V-FITC method utilizing flow cytometry. Amplitude modulated (217 pulses per second) RF exposure significantly enhanced UV induced apoptosis in cdc48-mutated cells, but no effect was observed in cells exposed to unmodulated fields at identical time-average specfic absorption rates (SAR, 0.4 or 3.0 W/kg). The findings suggest that amplitude modulated RF fields, together with known damaging agents, can affect the cell death process in mutated yeast cells. Bioelectromagnetics 25:127-133, 2004. PMID- 14735564 TI - Pulsed electromagnetic field stimulation of bone marrow cells derived from ovariectomized rats affects osteoclast formation and local factor production. AB - This study examined the effects of a specific pulsed electromagnetic field (PEMF) stimulation on osteoclast formation in bone marrow cells from ovariectomized rats and to determine if the signal modulates the production of cytokines associated with osteoclast formation. Adult female Wistar rats were subjected to bilateral or sham ovariectomy, and primary bone marrow cells were harvested at 4 days (Subgroup I) and 7 days (Subgroup II) after surgery. Primary bone marrow cells were subsequently placed in chamber slides and set inside solenoids powered by a pulse generator (300 micros, 7.5 Hz) for 1 h per day for 9 days (OVX + PEMF group). Others (INT, SHAM, and OVX groups) were cultured under identical conditions, but no signal was applied. Recruitment and authentication of osteoclast-like cells were evaluated by determining multinuclear, tartrate resistant acid phosphatase (TRAP) positive cells on day 10 of culture and by pit formation assay, respectively. The PEMF signal caused significant reductions in osteoclast formation in both Subgroups I (-55%) and II (-43%). Tumor necrosis factor-alpha (TNF-alpha), interleukin 1beta (IL-1beta), and interleukin 6 (IL-6) in OVX + PEMF group of Subgroup I were significantly reduced at 5, 7, and 9 days as compared to OVX group. The results found in this study suggest that osteoclastogenesis can be inhibited by PEMF stimulation, putatively due to a concomitant decrease in local factor production. Bioelectromagnetics 25:134-141, 2004. PMID- 14735565 TI - Mobile phones and children: is precaution warranted? AB - Are there health related arguments to recommend that children limit their use of mobile telephones? The International Expert Group on Mobile Phones from the UK concluded so, but did not come up with convincing scientific data to back this statement. The Health Council of the Netherlands approached the problem by considering whether developmental arguments might be found, i.e., asking if there reason to believe that the heads of children are more susceptible to the electromagnetic fields emitted by mobile telephones than those of adults. It concluded that no major changes in head development occur after the second year of life that might point at a difference in electromagnetic susceptibility between children and adults. The Health Council therefore sees no reason to recommend limiting the use of mobile phones by children. PMID- 14735566 TI - Modified Gaussian-2 level investigation of the identity ion-pair SN2 reactions of lithium halide and methyl halide with inversion and retention mechanisms. AB - Identity ion-pair S(N)2 reactions LiX + CH(3)X --> XCH(3) + LiX (X = F, Cl, Br, and I) have been investigated in the gas phase and in solution at the level of the modified Gaussian-2 theory. Two possible reaction mechanisms, inversion and retention, are discussed. The reaction barriers relative to the complexes for the inversion mechanism [DeltaH(cent) ( not equal )(inv)] are found to be much higher than the corresponding values for the gas phase anionic S(N)2 reactions, decreasing in the following order: F (263.6 kJ mol(-1)) > Cl (203.3 kJ mol(-1)) > Br (174.7 kJ mol(-1)) > I (150.7 kJ mol(-1)). The barrier gaps between the two mechanisms [DeltaH(cent) ( not equal ) (ret) - DeltaH(cent) ( not equal ) (inv)] increase in the order F (-62.7 kJ mol(-1)) < Cl (4.4 kJ mol(-1)) < Br (24.9 kJ mol(-1)) < I (45.1 kJ mol(-1)). Thus, the retention mechanism is energetically favorable for fluorine and the inversion mechanism is favored for other halogens, in contrast to the anionic S(N)2 reactions at carbon where the inversion reaction channel is much more favorable for all of the halogens. The stabilization energies for the dipole-dipole complexes CH(3)X. LiX (DeltaH(comp)) are found to be similar for the entire set of systems with X = F, Cl, Br, and I, ranging from 53.4 kJ mol(-1) for I up to 58.9 kJ mol(-1) for F. The polarizable continuum model (PCM) has been used to evaluate the direct solvent effects on the energetics of the anionic and ion-pair S(N)2 reactions. The energetic profiles are found to be still double-well shaped for most of the ion-pair S(N)2 reactions in the solution, but the potential profile for reaction LiI + CH(3)I is predicted to be unimodal in the protic solvent. Good correlations between central barriers [DeltaH(cent) ( not equal ) (inv)] with the geometric looseness of the inversion transition state %C-X( not equal ), the dissociation energies of the C-X bond (D(C-X)) and Li-X bond (D(Li-X)) are observed, respectively. PMID- 14735567 TI - A practical valence bond method: a configuration interaction method approach with perturbation theoretic facility. AB - The previously developed valence bond configuration interaction (VBCI) method (Wu, W.; Song, L.; Cao, Z.; Zhang, Q.; Shaik, S., J. Phys. Chem. A, 2002, 105, 2721) that borrows the general CI philosophy of the MO theory, is further extended in this article, and its methodological features are improved, resulting in three accurate and cost-effective procedures: (a) the effect of quadruplet excitation is incorporated using the Davidson correction, such that the new procedure reduces size consistency problems, with due improvement in the quality of the computational results. (b) A cost-effective procedure, named VBCI(D, S), is introduced. It includes doubly excited structures for active electrons and singly excited structures for inactive pairs. The computational results of VBCI(D, S) match those of VBCISD with much less computational effort than VBCISD. (c) Finally, a second-order perturbation theory is utilized as a means of configuration selection, and lead to considerable reduction of the computational cost, with little or no loss in accuracy. Applications of the new procedures to bond energies and barriers of chemical reactions are presented and discussed. PMID- 14735568 TI - AGBNP: an analytic implicit solvent model suitable for molecular dynamics simulations and high-resolution modeling. AB - We have developed an implicit solvent effective potential (AGBNP) that is suitable for molecular dynamics simulations and high-resolution modeling. It is based on a novel implementation of the pairwise descreening Generalized Born model for the electrostatic component and a new nonpolar hydration free energy estimator. The nonpolar term consists of an estimator for the solute-solvent van der Waals dispersion energy designed to mimic the continuum solvent solute solvent van der Waals interaction energy, in addition to a surface area term corresponding to the work of cavity formation. AGBNP makes use of a new parameter free algorithm to calculate the scaling coefficients used in the pairwise descreening scheme to take into account atomic overlaps. The same algorithm is also used to calculate atomic surface areas. We show that excellent agreement is achieved for the GB self-energies and surface areas in comparison to accurate, but much more expensive, numerical evaluations. The parameter-free approach used in AGBNP and the sensitivity of the AGBNP model with respect to large and small conformational changes makes the model suitable for high-resolution modeling of protein loops and receptor sites as well as high-resolution prediction of the structure and thermodynamics of protein-ligand complexes. We present illustrative results for these kinds of benchmarks. The model is fully analytical with first derivatives and is computationally efficient. It has been incorporated into the IMPACT molecular simulation program. PMID- 14735569 TI - Continuous medium theory for nonequilibrium solvation: I. How to correctly evaluate solvation free energy of nonequilibrium. AB - Considering the influences of electrostatic potential Phi upon the change of solute charge distribution deltarho and rho upon the change deltaPhi at the same time, a more reasonable integral formula of dG = (1/2) integral (V) (rhodeltaPhi + Phideltarho)dV is used to calculate the change of the electrostatic free energy in charging the solute-solvent system to a nonequilibrium state, instead of the one of dG = integral (V) PhideltarhodV used before. This modification improves the expressions of electrostatic free energy and solvation free energy, in which no quantity of the intermediate equilibrium state is explicitly involved. Detailed investigation reveals that the solvation free energy of nonequilibrium only contains the interaction energy between the field due to the solute charge in vacuum, and the dielectric polarization at the nonequilibrium state. The solvent reorganization energies of forward and backward electron transfer reactions have been redefined because the derivations lead to a remarkable feature that these quantities are direction-dependent, unlike the theoretical models developed before. The deductions are given in the electric field displacement form. Relevant discussions on the reliability of theoretical models suggested in this work have also been presented. PMID- 14735570 TI - A kinematic view of loop closure. AB - We consider the problem of loop closure, i.e., of finding the ensemble of possible backbone structures of a chain segment of a protein molecule that is geometrically consistent with preceding and following parts of the chain whose structures are given. We reduce this problem of determining the loop conformations of six torsions to finding the real roots of a 16th degree polynomial in one variable, based on the robotics literature on the kinematics of the equivalent rotator linkage in the most general case of oblique rotators. We provide a simple intuitive view and derivation of the polynomial for the case in which each of the three pair of torsional axes has a common point. Our method generalizes previous work on analytical loop closure in that the torsion angles need not be consecutive, and any rigid intervening segments are allowed between the free torsions. Our approach also allows for a small degree of flexibility in the bond angles and the peptide torsion angles; this substantially enlarges the space of solvable configurations as is demonstrated by an application of the method to the modeling of cyclic pentapeptides. We give further applications to two important problems. First, we show that this analytical loop closure algorithm can be efficiently combined with an existing loop-construction algorithm to sample loops longer than three residues. Second, we show that Monte Carlo minimization is made severalfold more efficient by employing the local moves generated by the loop closure algorithm, when applied to the global minimization of an eight-residue loop. Our loop closure algorithm is freely available at http://dillgroup. ucsf.edu/loop_closure/. PMID- 14735571 TI - Partial rigid-body dynamics in NPT, NPAT and NPgammaT ensembles for proteins and membranes. AB - A partial rigid-body method of molecular dynamics simulations for proteins and membranes is presented. In this method, the symplectic integrator for rigid bodies is combined with the equations of motion for the NPT ensemble. The standard NPT ensemble is extended to the membrane-specific ensembles, the NPAT (constant normal pressure and lateral surface area of membranes and constant temperature) and NPgammaT (constant normal pressure and lateral surface tension of membranes and constant temperature) ensembles. By more than 30-ns simulations of aqueous proteins and hydrated lipid bilayers, the results of the partial rigid body method demonstrated excellent conservation of total energy and consistent behavior with the traditional constraint method in terms of structural distribution and fluctuation of proteins and lipids. The efficient implementation of the partial rigid-body method in parallel computation is presented, which is shown to work well in large-scale molecular dynamics simulations. PMID- 14735572 TI - An AM1 semiempirical study of host-guest complexation in hemicarcerand complexes. AB - Semiempirical AM1 calculations have been carried out on host-guest complexes of model hemicarcerands 1a and 2a. The justification for the choice of the AM1 Hamiltonian was based on a comparison between reported X-ray data for the smaller tetrabromocavitand 4a and computational results obtained using several different Hamiltonians. The complexation behavior of hemicarcerands 1a and 2a have been compared with experimental results reported by Cram et al. for the related hemicarcerands 1b and 2b. Based on this comparison, a criterion for predicting guest encapsulation was developed, E(complexation), which relies on the calculation of AM1 heats of formation for host, guest, and hemicarceplex. If E(complexation) is lower than 10 kcal/mol, then a guest will be encapsulated, while if it is greater than 30 kcal/mol, a guest will not be encapsulated. The use of constrained-path AM1 optimizations to determine the energy barriers to guest entry and exit from the host was found to be a useful tool for examining suitable host-guest combinations when the E(complexation) criteria does not hold. We have computed the barriers to exit of N, N-dimethylformamide (dmf) and furan from the hemicarcerand 1a, the former has been compared with the experiment and shows excellent agreement. Based on the success of the above computational methods in predicting which host-guest combinations will form stable hemicarceplexes we have synthesized a new target hemicarceplex 1b.furan. PMID- 14735573 TI - Direct ab initio dynamics studies on the hydrogen-abstraction reactions of OH radicals with HOX (X = F, Cl, and Br). AB - The hydrogen abstract reactions of OH radicals with HOF (R1), HOCl (R2), and HOBr (R3) have been studied systematically by a dual-level direct-dynamics method. The geometries and frequencies of all the stationary points are optimized at the MP2/6-311+G(2d, 2p) level of theory. A hydrogen-bonded complex is located at the product channel for the OH + HOBr reaction. To improve the energetics information along the minimum energy path (MEP), single-point energy calculations are carried out at the CCSD(T)/6-311++G(3df, 3pd) level of theory. Interpolated single-point energy (ISPE) method is employed to correct the energy profiles for the three reactions. It is found that neither the barrier heights (DeltaE) nor the H-O bond dissociation energies [D(H-O)] exhibit any clear-cut linear correlations with the halogen electronegative. The decrease of DeltaE and D(H-O) for the three reactions are in order of HOF > HOBr > HOCl. Rate constants for each reaction are calculated by canonical variational transition-state theory (CVT) with a small curvature tunneling correction (SCT) within 200-2000 K. The agreement of the rate constants with available experimental values for reactions R2 and R3 at 298 K is good. Our results show that the variational effect is small while the tunneling correction has an important contribution in the calculation of rate constants in the low-temperature range. Due to the lack of the kinetic data of these reactions, the present theoretical results are expected to be useful and reasonable to estimate the dynamical properties of these reactions over a wide temperature range where no experimental value is available. PMID- 14735574 TI - Conformational search of peptides and proteins: Monte Carlo minimization with an adaptive bias method applied to the heptapeptide deltorphin. AB - The energy function of a protein consists of a tremendous number of minima. Locating the global energy minimum (GEM) structure, which corresponds approximately to the native structure, is a severe problem in global optimization. Recently we have proposed a conformational search technique based on the Monte Carlo minimization (MCM) method of Li and Scheraga, where trial dihedral angles are not selected at random within the range [-180 degrees,180 degrees ] (as with MCM) but with biased probabilities depending on the increased structure-energy correlations as the GEM is approached during the search. This method, called the Monte Carlo minimization with an adaptive bias (MCMAB), was applied initially to the pentapeptide Leu-enkephalin. Here we study its properties further by applying it to the larger peptide with bulky side chains, deltorphin (H-Tyr-D-Met-Phe-His-Leu-Met-Asp-NH(2)). We find that on average the number of energy minimizations required by MCMAB to locate the GEM for the first time is smaller by a factor of approximately three than the number required by MCM-in accord with results obtained for Leu-enkephalin. PMID- 14735575 TI - A hydration study of (1-->4) and (1-->6) linked alpha-glucans by comparative 10 ns molecular dynamics simulations and 500-MHz NMR. AB - The hydration behavior of two model disaccharides, methyl-alpha-D-maltoside (1) and methyl-alpha-D-isomaltoside (2), has been investigated by a comparative 10 ns molecular dynamics study. The detailed hydration of the two disaccharides was described using three force fields especially developed for modeling of carbohydrates in explicit solvent. To validate the theoretical results the two compounds were synthesized and subjected to 500 MHz NMR spectroscopy, including pulsed field gradient diffusion measurements (1: 4.0. 10(-6) cm(2). s(-1); 2: 4.2. 10(-6) cm(2). s(-1)). In short, the older CHARMM-based force field exhibited a more structured carbohydrate-water interaction leading to better agreement with the diffusional properties of the two compounds, whereas especially the alpha-(1- >6) linkage and the primary hydroxyl groups were inaccurately modeled. In contrast, the new generation of the CHARMM-based force field (CSFF) and the most recent version of the AMBER-based force field (GLYCAM-2000a) exhibited less structured carbohydrate-water interactions with the result that the diffusional properties of the two disaccharides were underestimated, whereas the simulations of the alpha-(1-->6) linkage and the primary hydroxyl groups were significantly improved and in excellent agreement with homo- and heteronuclear coupling constants. The difference between the two classes of force field (more structured and less structured carbohydrate-water interaction) was underlined by calculation of the isotropic hydration as calculated by radial pair distributions. At one extreme, the radial O em leader O pair distribution function yielded a peak density of 2.3 times the bulk density in the first hydration shell when using the older CHARMM force field, whereas the maximum density observed in the GLYCAM force field was calculated to be 1.0, at the other extreme. PMID- 14735576 TI - Vibrational center-ligand couplings in transition metal complexes. AB - The mode-tracking principle [J. Chem. Phys. 2003, 118, 1634] for the direct quantum chemical calculation of preselected, characteristic molecular vibrations makes vibrational analyses of very large molecules feasible. This is demonstrated here for the [(Ph(3)PAu)(6)C](2+) complex, in which 18 phenyl groups in the ligand sphere are explicitly taken into account. We are aiming at the motion of the endohedral carbon atom, which is in an extraordinary bonding situation because it is surrounded by an octahedral core of gold atoms in this cluster. Secondary effects of the full ligand sphere on the vibrations of the [Au(6)C] core embedded in [(R(3)PAu)(6)C](2+) clusters are investigated. For this purpose, local vibrations of the octahedral core are generated, and their long-range couplings with the phosphine ligand sphere become visible in the mode-tracking iterations. The exact normal modes of these characteristic vibrations of the cluster are then obtained after convergence of the mode-tracking refinement. This protocol allows us to assess the coupling of the outer ligand sphere with the inner core of the cluster in terms of changes of the vibrational frequencies and of the collective motions of the atomic nuclei. The vibrational frequencies of the octahedral [Au(6)C] core split due to symmetry breaking in the C(1)-symmetric [(Ph(3)PAu)(6)C](2+) cluster. Our study demonstrates how effects of the periphery of a large molecule on local vibrations can be quantified. Furthermore, we predict the first set of characteristic vibrational frequencies obtained with first-principles methods for this gold cluster, whose vibrational spectra have not yet been recorded experimentally. PMID- 14735577 TI - A computational study of the role of hydrogen bonds in SN1 and E1 reactions. AB - The reaction between tertiary butyl chloride and water clusters was examined by applying density functional theory calculations. The carbonium ion t-Bu(+) that is normally sandwiched between the water clusters was found to be absent, such that a Cbond;O covalent bond was formed in the intermediate (Int1) after heterolysis. An (H(2)O)(4) cluster is able to bridge the front and rear of the central carbon and promotes heterolysis. A correlation between bond interchanges at the central carbon and proton relays is presented. Stereochemical scrambling in the solvolysis products is discussed in terms of this correlation. In addition, an E1 pathway for the elimination product, iso-butene, is found from Int1. PMID- 14735578 TI - Accurate and efficient generalized born model based on solvent accessibility: derivation and application for LogP octanol/water prediction and flexible peptide docking. AB - A novel method for fast and accurate evaluation of the generalized Born radii in macromolecular solvation electrostatics calculations is proposed, based on the solvent accessibility of the first two solvation layers around an atom. The reverse generalized Born radii calculated by the method have correlation coefficient of 98.7% and RMSD of 0.031 A(-1) with the values obtained using a precise but significantly slower numerical boundary element solution. The method is applied to derive an estimate of the free solvation energy difference between octanol and water and to predict LogP octanol-water. A nine-parameter model is optimized on an 81 compound training set and applied to predict LogP(ow) for an external evaluation set of 19 drug molecules with RMSD of 0.9. The new GB approximation is also tested in Monte Carlo docking simulations of the fully flexible p53 peptide fragment to MDM2. The best energy solution found in the simulations has RMSD of 2.8 A to the X-ray structure. PMID- 14735579 TI - Prader-Willi syndrome: causes of death in an international series of 27 cases. AB - Prader-Willi syndrome (PWS) is a complex condition with many medical and psychological features. In individuals with this syndrome, causes of death were studied. Data of 27 case reports were collected. Ages at death ranged from neonatal to 68 years. None of the individuals were treated with growth hormone (GH). Most cases were not completely documented and autopsy was performed in a minority of cases only. In five cases, death was considered not to be causally related to PWS. Hypotonia with hypoventilation was noted in the babies, and acute respiratory illness with unexpected sudden death was experienced in young children with PWS. Two young children died after a short period of fever and gastroenteritis. Obesity and its complications leading to death were pronounced in the adult group. One (possibly two) adult(s) died from gastric dilatation and shock. Based on these data, some cautious conclusions can be drawn. In babies with PWS hypoventilation is a risk factor; upper airway infection may be more serious than anticipated and any other clinical features pointing to an infection should be taken very seriously. Therefore, young infants with PWS hospitalized with an upper airway infection and/or hypoventilation or gastroenteritis symptoms, should be closely monitored. Early diagnosis and prevention of overweight is a major factor in preventing early causes of death in individuals with PWS. In the adult group, weight reduction is important but difficult to manage. Sleep apnea should be recognized and treated. Pain in the upper stomach and/or vomiting should be taken as a possible sign of acute intestinal dilatation; intravenous support may be life saving. PMID- 14735580 TI - New 19 bp deletion polymorphism in intron-1 of dihydrofolate reductase (DHFR): a risk factor for spina bifida acting in mothers during pregnancy? AB - Up to 72% of spina bifida cystica (SB) is preventable by maternal periconceptual folic acid supplementation. The C677T allele of the methylenetetrahydrofolate reductase (MTHFR) gene and some other functional polymorphisms are risk factors for SB in some populations. However, despite extensive study, the genetic risk factors for SB are incompletely understood. Polymorphic alleles that diminish bioavailability of reduced folate in the mother during pregnancy could contribute to SB in her fetus, acting in the mother as teratogenic alleles. We recently discovered a polymorphic 19 bp deletion allele (frequency 0.45) within intron-1 of dihydrofolate reductase (DHFR) that is a good candidate for such a genetic factor. Since there is precedence for intron-1 regulatory elements and the deletion allele removes a potential Sp1 transcription factor binding site, we hypothesized that the deletion allele could be functional and act in SB mothers to increase the risk of SB in her fetus. We found that homozygosity for this deletion allele was significantly more frequent in SB mothers, but not in SB fathers or patients, compared with controls and was associated with a significantly increased odds ratio (OR) (2.035) of being an SB mother compared with other genotypes. Genotype distribution obeyed the constraints of Hardy Weinberg equilibrium in controls, SB patients and fathers, but not in SB mothers. If confirmed, these findings could lead to improved forms of folate supplementation for pregnancy. About half of dietary folates and all of folic acid supplements must be reduced by DHFR to be available for mother and fetus. Reduced folates could be preferable for supplements during pregnancy to prevent SB. PMID- 14735581 TI - Impact of BRCA1/2 testing and disclosure of a positive test result on women affected and unaffected with breast or ovarian cancer. AB - To evaluate the impact of BRCA1/2 testing and disclosure of a positive test result on women affected and unaffected with cancer. Longitudinal cohort study including women affected and unaffected with breast or ovarian cancer testing for a BRCA1/2 mutation. Data on well-being (anxiety, depression, cancer related distress, general health), treatment choice, and decision making about cancer prevention were collected at baseline (1 week after blood sampling; affected n = 192, unaffected n = 176) and at follow-up (2 weeks after disclosure of a positive test result; affected n = 23, unaffected n = 66). Women affected and unaffected with breast or ovarian cancer were compared using univariate statistics. Change over time was examined using repeated measures analysis of variance. With respect to well-being, affected women scored worse at baseline. At follow-up, both affected and unaffected women experienced a decline in well-being, which tended to be stronger in affected women. Women diagnosed with cancer less than 1 year previously tended to report a worse well-being than those diagnosed longer ago. With respect to treatment choice, more affected women intended to obtain prophylactic surgery and valued it higher at both time points. With respect to decision making, affected women had a lower preference for participation in decision making at baseline; no differences were found at follow-up. At follow up, both affected and unaffected women showed an increase in strength of treatment preference and a decrease in decision uncertainty. Disclosure of a positive test result had a negative impact on well-being. Affected women, especially those who have been recently diagnosed with cancer, experienced the worst well-being and could benefit from psychosocial support. PMID- 14735582 TI - Brachydactyly type C caused by a homozygous missense mutation in the prodomain of CDMP1. AB - Brachydactyly type C (BDC) is characterized by shortening of the middle phalanges of the index, middle, and little finger with hyperphalangy, usually of the index and middle finger. Heterozygous mutations of the cartilage derived morphogenetic protein-1 (CDMP1) resulting in a loss of function have been reported in BDC. We here describe a large kindred with a semi-dominant form of BDC and pronounced ulnar deviation of the second and third digits. In this family a novel homozygous missense mutation was identified (517A > G) changing methionine to valine at amino acid position 173. The mutation is located within a highly conserved seven amino acid region of the prodomain of CDMP1. Hand radiographs of heterozygous mutation carriers showed mild shortening of the metacarpals IV and V; a finding confirmed by the analysis of their metacarpophalangeal profiles (MCPPs). The mutation described here points toward an important function of the prodomain for the folding, secretion, and availability of biologically active CDMP1. PMID- 14735583 TI - Specific clinical and brain MRI features in mentally retarded patients with mutations in the Oligophrenin-1 gene. AB - Oligophrenin-1 (OPHN-1) gene disruption is known as responsible for so called "non-specific" X-linked mental retardation (MR) Billuart et al. [1998: Nature 392:923-926]. In order to search for a possible specific clinical and radiological profile for mutation in the OPHN-1 gene, clinical and 3D brain MRI studies were performed in the two families with a known mutation in OPHN-1 reported so far: a 19-year-old female with an X;12 balanced translocation encompassing OPHN-1, and four affected males of family MRX60 sharing a frameshift mutation in OPHN-1. Clinical data shared by affected individuals were neonatal hypotonia with motor delay but no obvious ataxia, marked strabismus, early onset complex partial seizures, and moderate to severe MR. Brain MRIs performed in three individuals exhibited a specific vermian dysgenesis including an incomplete sulcation of anterior and posterior vermis with the most prominent defect in lobules VI and VII. In addition, a non-specific cerebral cortico-subcortical atrophy was also observed. These clinical and radiological features suggest a distinct clinico-radiological syndrome. These preliminary data need to be confirmed in other families and will be helpful for further targeted mutation screening of the OPHN-1 gene in male patients with similar clinico-radiological features. In addition, OPHN-1 inactivation should be considered as a relevant model of developmental vermis disorganization, leading to a better understanding of the possible role of the cerebellum in MR. PMID- 14735584 TI - Clinical variability in maternally inherited leber hereditary optic neuropathy with the G14459A mutation. AB - Spasticity and dystonia have been associated with mitochondrial (mt) DNA mutations at A11696G, G14459A, and T14596A. We describe the clinical features and molecular analysis of two Caucasian pedigrees with the 14,459 guanosine (G) --> adenine (A) transition. The maternally inherited Leber hereditary optic neuropathy (LHON) phenotypes showed extreme clinical variability and the only screening test that was abnormal in the patient with spasticity/dystonia was a high T2 signal in the putamen bilaterally. The male patient in the second pedigree showed features of optic neuropathy without spasticity/dystonia. These results further support that the 14,459 G --> A transition mutation is causally related to LHON and spasticity/dystonia. PMID- 14735585 TI - Variable clinical manifestation of homoplasmic G14459A mitochondrial DNA mutation. AB - Leber hereditary optic neuropathy (LHON)/pediatric onset dystonia is associated with a G to A transition at nucleotide position (np) 14459, within the mitochondrial DNA (mtDNA)-encoded ND6 gene. This mutation has been reported in families presenting with LHON alone, LHON plus dystonia, or pediatric dystonia with typical age of onset less than 5 years. The mutation changes a moderately conserved alanine to a valine at amino acid residue 72, which is within the most evolutionarily conserved region of the ND6 protein. Pediatric onset disease is associated with basal ganglia dysfunction, spasticity, and encephalopathy. We report a family with G14459A mtDNA mutation and a broad spectrum of clinical manifestation. The proband was a 3-year-old girl with anarthria, dystonia, spasticity, and mild encephalopathy. MRI of the brain demonstrated bilateral, symmetric basal ganglia lucencies associated with cerebral and systemic lactic acidosis. Her maternal first cousin presented with a new onset limp and mild hemiparesis along with similar MRI findings with a much milder phenotype. Additional investigation of the family members with the mutation has revealed both asymptomatic and symptomatic individuals with variable clinical and laboratory features of mitochondrial disease. This study re-emphasizes the heterogeneous clinical manifestation of homoplasmic G14459A mtDNA mutation even within the same family, and supports the hypothesis that nuclear genes may play a role in modifying the clinical expression of mitochondrial disease. Published 2003 Wiley-Liss, Inc. PMID- 14735586 TI - Constitutional trisomy 8 mosaicism due to meiosis II non-disjunction in a phenotypically normal woman with hematologic abnormalities. AB - Constitutional trisomy 8 mosaicism (CT8M) in liveborns is typically caused by mitotic non-disjunction and exhibits wide phenotypic variability. By contrast, CT8M due to meiotic errors usually results in miscarriage. We describe a case of CT8M due to a paternal meiosis II non-disjunction error. The patient, a 32-year old woman, was phenotypically normal except for a history of recurrent aphthous ulcers since childhood and a 4-year history of macrocytosis. The ulcers were refractory to steroids, but responded well to thalidomide. To the best of our knowledge, this is the first report of CT8M due to meiotic non-disjunction in a phenotypically normal individual. PMID- 14735587 TI - Growth hormone deficiency (GHD): a new association in Peters' Plus Syndrome (PPS). AB - We report a case of a girl with clinical features of Peters' Plus Syndrome (PPS) (association of anterior eye chamber defects; peculiar facies; cleft lip/palate; brachymelia; developmental delay; growth retardation) and documented growth hormone deficiency (height -3.5 SDS at chronological age 5 years 8 months; low growth factors; bone age delay; growth velocity 4.4 cm/year (<3rd centile); and peak growth hormone levels of 1.7 and 4.7 ng/ml by clonidine and insulin provocative testing, respectively). Treatment with recombinant human growth hormone (0.3 mg/kg/week) resulted in a dramatic increase in growth velocity, increasing the height from -3.5 to -1.5 SDS over 2.3 years of therapy, indicative of an excellent response. Growth retardation is a known association in PPS: a condition that includes other midline facial defects. This case supports a role for GHD in the pathogenesis of the short stature observed in these children; demonstrates the efficacy of GH treatment; and further reinforces the relationship of pituitary anomalies with common congenital defects. PMID- 14735589 TI - Retinoblastoma, pinealoma, and mild overgrowth in a boy with a deletion of RB1 and neighbor genes on chromosome 13q14. AB - We report on a 10-year-old boy with a normal karyotype and a chromosome 13q14 deletion of the retinoblastoma gene (RB1) by fluorescence in situ hybridization (FISH). He showed subtle signs of overgrowth, including macrocephaly, hepatomegaly, and inguinal hernia. The boy also had cryptorchism and mild developmental delay. In his first months of life, variant Wiedemann-Beckwith syndrome was tentatively suspected and he was included in a careful tumor prevention program. At the age of 11 months, bifocal retinoblastoma of the left eye was diagnosed. Pinealoma was suspected at the age of 19 months and was removed by neurosurgery at the age of 29 months. At 4 years and 4 months, the deletion of the RB1 gene was suspected on clinical grounds and was diagnosed by FISH and molecular studies. At that time, he was a near-normal healthy playful kindergarten child, height 107 cm (-0.3 SD), OFC 52.5 cm (+0.8 SD), developmental age 3-3.5 years. The combination of retinoblastoma, pinealoma, and deletion of the RB1 gene diagnosed by FISH has not been reported previously. The deletion spans at least 370-420 kb in size and is predicted to include proximal and distal neighbor genes. This report may assist in establishing the clinical signs of the contiguous gene syndrome at the RB1 locus on 13q14. PMID- 14735588 TI - Spinocerebellar ataxia type 2 (SCA2) presenting with ophthalmoplegia and developmental delay in infancy. AB - An 11-year-old boy was evaluated for progressive ataxia, cognitive deterioration, and ophthalmoplegia. The child initially presented with abnormal eye movements at the age of 2 months and was noted to have developmental delay at 6 months. At the age of 7 years, he developed ataxia and cognitive impairment, and subsequently manifested dysphagia and incontinence. The pertinent family history included gait difficulty in the paternal grandmother. At the age of 11, his general physical examination was normal. On neurological examination, he had bilateral external ophthalmoplegia, ataxic dysarthria, dysmetria and tremor in the upper extremities, and marked gait ataxia. An ophthalmological evaluation showed no evidence of pigmentary retinopathy. Brain MRI demonstrated cerebellar, brainstem, and cerebral atrophy. An ataxia panel showed 62 repeats in one allele of the SCA2 gene. Most cases of spinocerebellar ataxia type 2 (SCA2) present between 20 years and 40 years, and affected individuals typically have between 34 and 57 CAG repeats. Neonatal cases of SCA2 have been reported in individuals with over 200 CAG repeats. Childhood SCA2 has been reported previously in two patients but not described clinically. This case broadens the spectrum of the clinical features of infantile-onset SCA2 and highlights the importance of considering this diagnosis in infants and children. PMID- 14735590 TI - Two cases of tetrasomy 9p syndrome with tissue limited mosaicism. AB - Tetrasomy of short arm of chromosome 9 constitutes a clinically recognizable chromosomal syndrome. Isochromosome 9p shows a strong propensity to tissue limited mosaicism. It occurs predominantly in peripheral blood cultures, often at a lower frequency or even absent in skin, amniotic fluid or chorionic villous cell cultures. Tissue-limited nature of mosaicism may render prenatal detection of this condition very difficult. Herein, we report two new cases of mosaic tetrasomy 9p. Conventional cytogenetics (CC) and FISH studies demonstrated a differential expression of the mosaicism in several tissues. We review the literature and discuss the implications of these findings in cytogenetic prenatal diagnosis. PMID- 14735591 TI - A family with X-linked recessive fusion of metacarpals IV and V. AB - We describe a family with a distinctive malformation of the hand consisting of the fusion of the 4th and the 5th metacarpal bones. Usually this anomaly is clinically recognizable by an ulnar deviation of the 5th finger; moreover, the 5th metacarpal is usually hypoplastic and the 5th ray is consequently short. There is, however, great variability in expression, so the degree of fusion may range from minimal to complete and also the external aspect of the hand may vary. This anomaly can be either isolated or part of a syndrome. For the isolated form, two possible hereditary mechanisms have been proposed: autosomal dominant and X linked recessive. Our family is consistent with the latter, with only affected males and no instances of male-to-male transmission. Of note, there are very few X-linked recessive disorders that affect the hand in a such a specific way. PMID- 14735592 TI - Maternal origin of a de novo mutation of the connexin 26 gene resulting in recessive nonsyndromic deafness. PMID- 14735593 TI - Does the P172H mutation at the TM4SF2 gene cause X-linked mental retardation? PMID- 14735594 TI - Down syndrome adults. PMID- 14735595 TI - Pre- and postnatal diagnosis of limb anomalies: a series of 107 cases. AB - This is a 3-year retrospective study of 107 cases presenting with limb anomalies detected either on prenatal ultrasound scan, or after birth. These limb malformations are developmental anomalies, and can be isolated, syndromic, or associated with multiple malformations. Cases were ascertained through the prenatal diagnosis center, the pediatrics department, and the feto-pathology department. Several criteria were analyzed including sex ratio, prenatal diagnosis, karyotype, termination of pregnancies, clinical or pathological examination, pediatric or surgical and/or genetic assessment, and whether or not a diagnosis was made. Positional deformities and syndactyly were excluded. Limb anomalies were detected prenatally in 45% of the cases, and a diagnosis was made in 78%, including isolated, syndromic, or chromosomal anomalies. Sixty-one per cent of the infants had follow-up, either pediatric, surgical, or genetic. Prenatal multidisciplinary assessment is fundamental to assist with counseling, as is the post-natal follow-up of the infant. The diagnosis, if made, will obviously influence the information that will be given to the parents and the management of the malformation. If the pregnancy is terminated, feto-pathological examination is essential to help make a diagnosis, and guide recurrence risks. We are currently undertaking a prospective study, and we will develop a protocol of investigations in the future, depending on the type of the malformation identified. PMID- 14735596 TI - Antenatal manifestations of Smith-Lemli-Opitz (RSH) syndrome: a retrospective survey of 30 cases. AB - Smith-Lemli-Opitz (SLO) syndrome or RSH syndrome is an autosomal recessive multiple malformation, and mental retardation syndrome ascribed to 7 dehydrocholesterol reductase deficiency, and usually diagnosed in the early postnatal period. Reviewing a series of 30 cases of SLO, we have investigated the variable antenatal expression of the disorder. Intrauterine growth retardation (IUGR) was the most frequent detectable trait (20/30). IUGR was either isolated (9/20) or associated with at least one other anomaly (11/20), including nuchal edema, renal, cardiac, cerebral malformations, genital anomalies, or polydactyly. In this last group, 3/11 presented with multiple malformations (> or =3 anomalies). In 5/30 cases, isolated nuchal edema (3/30), and isolated cardiac (1/30) or renal malformations (1/30) were the only detectable anomalies. Ultrasound findings were considered normal in 5/30 cases and were abnormal in 25/30 cases (83%), but early detection of multiple malformations was rare (3/30, 10%). We suggest giving consideration to a more systematic sterol analysis when dealing with IUGR, especially when associated anomalies are detected. PMID- 14735597 TI - Fryns syndrome: a review of the phenotype and diagnostic guidelines. AB - Fryns syndrome (FS) is the commonest autosomal recessive syndrome associated with congenital diaphragmatic hernia (CDH) and comprises CDH, pulmonary hypoplasia, craniofacial anomalies, distal limb hypoplasia, and internal malformations. Although there have been more than 50 case reports on probands with FS, the diagnostic guidelines were formulated from a review of eight patients and modifications to the guidelines have only once been suggested. Recently, several case reports have described new anomalies in FS and other papers have highlighted the variation in expressivity found in FS. This paper examines the medical literature on FS to define the phenotype and to review the diagnostic guidelines. We conclude that CDH with brachytelephalangy and/or nail hypoplasia is strongly suggestive of the diagnosis and that pulmonary hypoplasia, craniofacial dysmorphism, orofacial clefting, and polyhydramnios are sufficiently frequent to be diagnostically useful. Other distinctive malformations that are consistent with FS include ventricular dilatation or hydrocephalus, agenesis of the corpus callosum, neuronal or cerebellar heterotopias, abnormalities of the aorta, renal cysts, dilatation of the ureters, bicornuate uterus, renal dysplasia, proximal thumbs, and broad clavicles. PMID- 14735598 TI - Hypothyroidism in Down syndrome: screening guidelines and testing methodology. PMID- 14735602 TI - Applying hypnosis in dermatology. PMID- 14735603 TI - Difficulties in diagnosing and treating tinea in adults at the Department of Dermatology in Bialystok (Poland). AB - In the years 1981-2000, the department of dermatology at the Medical University of Bialystok, Poland, carried out a retrospective study of common difficulties in the diagnosis and treatment of tinea. The aim of this study was to assess the incidence of incorrect diagnosis and therapy of tinea and tinea incognito (TI) in the patients hospitalized at the institution over a 19-year period. Tinea was identified in 814 patients (4.3% of all patients). TI was diagnosed in 318 patients (39.1% of all patients with tinea). The most diagnostic-therapeutic problems were observed in the patients with tinea pedis, tinea cutis glabrae superficialis, intertrigo candidamycetica, tinea profunda cutis glabrae and tinea profunda barbae. The most common clinical isolates were T. rubrum, T. mentagrophytes, and Candida albicans. A high percentage of TI was present in comparison with all other tinea conditions. PMID- 14735604 TI - What's your assessment? PMID- 14735605 TI - The prevalence of latex allergy among health care workers in Bolu (Turkey). AB - Allergic reactions to latex have become a significant public health problem, particularly among health care workers (HCWs). The objective of this cross sectional study was to determine the prevalence of latex allergy and risk factors for latex sensitization among HCWs in Bolu in Turkey. The prevalence of latex allergy or hypersensitivity among HCWs was 19.1%. HCWs with atopic disease, contact dermatitis, and increased glove use were at increased risk of sensitization. Increased awareness of the problem by HCWs is important to identify those at risk. PMID- 14735606 TI - Acne keloidalis nuchae. PMID- 14735616 TI - Let's make 2004 a year for nursing. PMID- 14735607 TI - Stasis dermatitis in the patient with CHF. PMID- 14735617 TI - What the public say. PMID- 14735618 TI - Image makeover. PMID- 14735620 TI - Writing it up. PMID- 14735621 TI - A specialist adolescent self-harm service. AB - AIM: To investigate the scale of adolescent deliberate self-harm (DSH) presentations to accident and emergency (A&E) departments and minor injury units in Glasgow and to assess the success of the adolescent DSH service in accessing this group. METHOD: A retrospective case note analysis was carried out in collaboration with the Greater Glasgow Health Board, using a standardised proforma. The data collection focused on adolescents aged 12-17 years presenting with DSH during a ten-week period in 2002. The records of each presentation to A&E departments and minor injury units were examined, and follow-up medical records were obtained for those who had been admitted to a ward. RESULTS: There were 86 recorded presentations of adolescent DSH during the study period. No significant relationships between age, sex, form of self-harm or any other variable recorded were found. Over half of those presenting were admitted to a ward, the majority of whom were subsequently referred to the DSH service. Those treated solely in A&E tended not to be referred. CONCLUSION: The findings of the study show that there are a significant number of young people who self-harm, but that they receive no follow-up care. PMID- 14735622 TI - Mucocutaneous separation. PMID- 14735623 TI - Benign ovarian neoplasms. AB - Benign ovarian neoplasms have the capacity to undergo malignant change and are difficult to diagnose in the early stages. Although rarely life-threatening, they can cause patients considerable physical and psychological distress. This article explains the structure and function of the ovaries and why they sometimes undergo benign neoplastic change. PMID- 14735625 TI - Can nurses stop the rise of the hospital superbug? PMID- 14735624 TI - Blood supplies hit by vCJD fear. PMID- 14735626 TI - The nurses improving pain management. PMID- 14735627 TI - The health secretary's vision for 2004. PMID- 14735628 TI - A new approach to respiratory care. PMID- 14735629 TI - The role of nurses in reducing the use of tobacco. AB - Smoking is a major cause of morbidity and mortality in the UK, and the government has funded a range of local and national smoking cessation initiatives. Supporting people as they attempt to give up smoking requires specialist skills and knowledge, often provided by nurses who have received appropriate training. However, non-specialist nurses can also play a role in cutting smoking rates by encouraging patients to consider giving up and directing them towards specialist services. This article focuses on the role that hospital-based nurses can play in encouraging patients to give up smoking. PMID- 14735630 TI - What you need to know about ... kidney stones. PMID- 14735631 TI - The impact of environmental cleanliness on infection rates. AB - Much has been said in recent years about the lack of environmental cleanliness in hospitals. The House of Lords Select Committee (1998) talked about falling standards in hospital cleaning and, more recently, the government acknowledged in The NHS Plan (Department of Health, 2000) that hospitals were unacceptably dirty. Such comments may affect people's perceptions of hospitals and, rather than regarding them as safe environments that promote healing, they may be viewed as dangerous places that might cause vulnerable patients greater harm. While it is known that about 10 per cent of hospital inpatients have a nosocomial infection at any one time (Emmerson et al, 1996), there is uncertainty about whether there is a sound evidence base to support the theory that dirty hospitals cause infection. PMID- 14735633 TI - Managing the care of patients who have visual impairment. AB - An ageing population means that the incidence of people who are visually impaired will increase. However, extending the role of ophthalmic nurses will promote delivery of a more effective health service for these patients. Using Maslow's hierarchy of needs as a basis for addressing the care of patients with visual impairment is a means of ensuring that they receive high quality, appropriate care at the right time. PMID- 14735632 TI - Upper respiratory tract carriage and transmission of pneumococci. AB - Vaccines have been instrumental in preventing childhood morbidity as well as mortality. It is imperative that health professionals have all the necessary information about a vaccine before its introduction into the routine immunisation schedule. This article looks at the possible introduction of a new pneumococcal conjugate vaccine and reviews a longitudinal carriage study of pneumococcus in UK families to determine pneumococcal carriage rates. PMID- 14735634 TI - Identifying types of dyspepsia and treatment. PMID- 14735635 TI - More than just mending bones. PMID- 14735636 TI - Evidence-based pandemonium. PMID- 14735637 TI - Nurses and preventable back injuries. PMID- 14735638 TI - Professionalism. PMID- 14735639 TI - Professionalism. PMID- 14735640 TI - Professionalism. PMID- 14735641 TI - Professionalism. PMID- 14735642 TI - Professionalism. PMID- 14735643 TI - Arginine immunonutrition in critically ill patients: a clinical dilemma. AB - Commercial enteral nutritional formulas for enhancement of the immune system are widely used in critical care. Immunonutrition with arginine can enhance inflammatory and immunologic responses in animal models and in humans. Although clinical improvements in surgical patients have been reported, benefits in critically ill patients with systemic inflammatory response syndrome, sepsis, or organ failure are less clear. Recent meta-analyses on the use of immunonutrition with arginine in critically ill and surgical patients revealed methodological weaknesses in most published studies. Specifically, a meta-analysis indicated that critically ill patients with preexisting severe sepsis may have an increased mortality rate when fed an immunonutritional enteral formula that contains arginine. These findings brought about confusion and controversy over the use of immunonutritional formulas in subsets of critically ill patients. A review of the literature on the function of arginine, its effect on the immune system, its roles in immunonutrition and in the clinical outcomes of critically ill patients, and the implications for nursing practice indicated that the benefits of immunonutrition with arginine in critically ill patients are unproven and warrant further study. Until more information is available, nutritional support should focus primarily on preventing nutritional deficiencies rather than on immunomodulation. PMID- 14735644 TI - Oral care in the intensive care unit. PMID- 14735645 TI - Oral health and care in the intensive care unit: state of the science. AB - Oral health is influenced by oral microbial flora, which are concentrated in dental plaque. Dental plaque provides a microhabitat for organisms and an opportunity for adherence of the organisms to either the tooth surface or other microorganisms. In critically ill patients, potential pathogens can be cultured from the oral cavity. These microorganisms in the mouth can translocate and colonize the lung, resulting in ventilator-associated pneumonia. The importance of oral care in the intensive care unit has been noted in the literature, but little research is available on mechanical or pharmacological approaches to reducing oral microbial flora via oral care in critically ill adults. Most research in oral care has been directed toward patients' comfort; the microbiological and physiological effects of tooth brushing in the intensive care unit have not been reported. Although 2 studies indicated reductions in rates of ventilator-associated pneumonia in cardiac surgery patients who received chlorhexidine before intubation and postoperatively, the effects of chlorhexidine in reducing ventilator-associated pneumonia in other populations of critically ill patients or its effect when treatment with the agent initiated after intubation have not been reported. In addition, no evaluation of the effectiveness of pharmacological and mechanical interventions relative to each other or in combination has been published. Additional studies are needed to develop and test best practices for oral care in critically ill patients. PMID- 14735646 TI - Effects of acuity-adaptable rooms on flow of patients and delivery of care. AB - BACKGROUND: Delayed transfers of patients between nursing units and lack of available beds are significant problems that increase costs and decrease quality of care and satisfaction among patients and staff. OBJECTIVE: To test whether use of acuity-adaptable rooms helps solve problems with transfers of patients, satisfaction levels, and medical errors. METHODS: A pre-post method was used to compare the effects of environmental design on various clinical and financial measures. Twelve outcome-based questions were formulated as the basis for inquiry. Two years of baseline data were collected before the unit moved and were compared with 3 years of data collected after the move. RESULTS: Significant improvements in quality and operational cost occurred after the move, including a large reduction in clinician handoffs and transfers; reductions in medication error and patient fall indexes; improvements in predictive indicators of patients' satisfaction; decrease in budgeted nursing hours per patient day and increased available nursing time for direct care without added cost; increase in patient days per bed, with a smaller bed base (number of beds per patient days). Some staff turnover occurred during the first year; turnover stabilized thereafter. CONCLUSIONS: Data in 5 key areas (flow of patients and hospital capacity, patients' dissatisfaction, sentinel events, mean length of stay, and allocation of nursing productivity) appear to be sufficient to test the business case for future investment in partial or complete replication of this model with appropriate populations of patients. PMID- 14735647 TI - B-type natriuretic peptide: a diagnostic, prognostic, and therapeutic tool in heart failure. AB - B-type natriuretic peptide is a neurohormone secreted from the cardiac ventricles in response to ventricular stretch and pressure overload. It counteracts the vasoconstriction that occurs as a compensatory mechanism in heart failure. A new test for measuring plasma levels of B-type natriuretic peptide can help in the diagnosis and treatment of patients with congestive heart failure. Dyspnea associated with cardiac dysfunction is highly unlikely in patients with levels of the peptide less than 100 pg/mL. Whereas most patients with significant congestive heart failure have levels of the peptide greater than 400 pg/mL, in patients with levels of 100 to 400 pg/mL, left ventricular dysfunction without volume overload, pulmonary embolism, and cor pulmonale must be ruled out. Thus, incorporating measurement of B-type natriuretic peptide into clinical evaluation helps physicians and nurses diagnose heart failure more quickly, especially in patients who have multiple comorbid conditions. Elevated levels of B-type natriuretic peptide indicate a poor prognosis in terms of a higher mortality and more hospital readmissions. Levels of B-type natriuretic peptide could be used to guide therapy and discharge planning for patients admitted with decompensated heart failure. PMID- 14735648 TI - Time in bed after electrophysiological procedures (TIBS IV): a pilot study. AB - BACKGROUND: Electrophysiological studies of the heart became commonplace in the past decade. Like cardiac catheterizations, electrophysiological studies are often considered "same day" procedures; patients are admitted in the morning, undergo the procedure, recover for several hours while confined to bed, and then are discharged from the hospital. The requisite time in bed varies widely between institutions where electrophysiological studies are performed. Little has been published about the optimal time that patients should remain in bed. OBJECTIVE: To determine if the requisite time in bed could be safely reduced by 2 hours for patients recovering from electrophysiological studies done via a femoral venous approach. METHODS: An experimental-control group design was used. A total of 68 patients were randomized to 2 hours (n = 31) or 4 hours (n = 37) of bed rest. Groups were comparable in age and sex. Both groups were observed for 5 hours after the procedure. RESULTS: The incidence of bleeding did not differ significantly between the experimental and control groups. Bleeding occurred in only 1 patient. CONCLUSIONS: The required 4 hours of bed rest after an electrophysiological study done via a femoral vein approach can safely be reduced to 2 hours. Early ambulation has implications for decreasing the cost of nursing care after the procedure and decreasing length of hospital stay, thus optimizing utilization of beds for recovery. PMID- 14735649 TI - Comparison of 3 methods of detecting acute respiratory distress syndrome: clinical screening, chart review, and diagnostic coding. AB - BACKGROUND: Although the incidence of acute respiratory distress syndrome has been studied, few researchers have prospectively assessed the search tool used to identify cases. METHODS: For 5 months, all patients admitted to a medical intensive care unit in a teaching hospital were evaluated daily to determine whether criteria for acute respiratory distress syndrome were met, and physicians' progress notes and discharge summaries for these prospectively identified patients were reviewed for mention of the syndrome. Discharge forms were reviewed for the codes (International Classification of Diseases, Ninth Revision) specific to acute respiratory distress syndrome (518.82 or 518.85). RESULTS: Of 314 patients admitted, 65 prospectively met the criteria for acute respiratory distress syndrome. Of these 65 patients, 31 had acute respiratory distress syndrome mentioned in their progress notes, and 4 of the 31 were subsequently assigned a code of 518.82 or 518.85. Patients with a physician's notation for acute respiratory distress syndrome in their charts had a higher mortality (22/31 [71%]) than did the patients with no such notation (10/34 [29%]). This difference could not be accounted for by differences in length of stay, mean age, score on Acute Physiology and Chronic Health Evaluation III, or number of days in the unit before meeting the criteria. CONCLUSIONS: The incidence of acute respiratory distress syndrome is underestimated when based on either diagnostic coding or physicians' notes without testing of the accuracy of coding. Both physicians and medical record coding specialists may require training in use of terms related to acute respiratory distress syndrome. PMID- 14735650 TI - Using bispectral index monitoring to detect potential breakthrough awareness and limit duration of neuromuscular blockade. PMID- 14735651 TI - The significance of hypothermia in preserving ischemic myocardium. AB - The clinical use of mild hypothermia to preserve ischemic cardiac and cerebral tissue continues to grow in popularity. This is a result of the known fact that hypothermia reduces myocardial oxygen demands more than any other intervention. The Advanced Life Support (ALS) Task Force of the International Liaison Committee on Resuscitation (ILCOR) made the following recommendations a year ago, in October 2002: "Unconscious adult patients with spontaneous circulation after out of-hospital cardiac arrest should be cooled to 32 degrees C to 34 degrees C for 12 to 24 hours when the initial rhythm was VF," or in-hospital even when arrest is due to other rhythms. Therapeutic use of hypothermia is in progress. PMID- 14735652 TI - Electrolyte imbalances. PMID- 14735653 TI - Chemical substances in the workplace can cause hearing loss. PMID- 14735654 TI - Accessibility regulations and a universal design philosophy inspire the design process. PMID- 14735655 TI - How now, mad cow? PMID- 14735656 TI - Beyond the sixth sense. PMID- 14735657 TI - Beyond ephedra. Why the FDA's ban may lead users to look for other weight-loss stimulants. PMID- 14735658 TI - Retirement plan trends: where are we all headed? AB - Many trends and their underlying pressures foster or hinder the creation of adequate retirement income. Generally, these trends fall into three categories: demographic issues, retirement phase of life and changes in corporate approaches to retirement plans. In an effort to facilitate everyone's participation in creating more palatable retirement plan solutions, this article describes how the trends within those three categories affect retirement income adequacy. PMID- 14735659 TI - Retirement behavior and retirement plan designs: strategies to retain an aging workforce. AB - Certain retirement and retiree health plan design features trigger different retirement behaviors among top performers and low performers and among workers of different wage levels. However, previous research has revealed little of these effects. Using a unique data set, this article shows how specific design features affect workers' retirement behavior, allowing employers to focus on those most likely to influence the most productive workers to delay retirement. PMID- 14735660 TI - SSA disability--weighing evidence--treating physicians' opinions. Krogmeier v. Barnhart. PMID- 14735661 TI - Disclosure--fiduciary duty--primary care physician incentives. Horvath v. Keystone Health Plan East, Inc. PMID- 14735662 TI - Health plans--reimbursement and subrogation after Great-West Life v. Knudson. Bauhaus USA, Inc. v. Copeland. PMID- 14735663 TI - Health plans--subrogation and reimbursement after Great-West Life v. Knudson- Equitable Lien. In re Carpenter. PMID- 14735666 TI - Acknowledging intellectual debt. PMID- 14735667 TI - Comments on nurse-patient relationship. PMID- 14735668 TI - Self-efficacy in rheumatoid arthritis. PMID- 14735669 TI - Power in organizations. PMID- 14735670 TI - Measuring progress in nursing and midwifery globally. AB - PURPOSE: To discuss issues and challenges encountered in the preliminary steps of identifying a core set of global indicators for measuring progress in nursing and midwifery development. ORGANIZING CONSTRUCT: Lack of a global and systematic approach for collecting evidence has undermined the ability to reliably measure progress. METHODS: Description of general principles, scientific criteria, and strategic directions. CONCLUSIONS: WHO's organizational support and global conditions are forces that indicated appropriate timing for these strategic directions. Concerted efforts are needed in all countries to implement these directions to advance nursing and midwifery services. PMID- 14735671 TI - Analysis and commentary on WHO Strategic Directions for Nursing and Midwifery. PMID- 14735672 TI - Commentary regarding the Sub-Saharan African region. PMID- 14735673 TI - An international perspective on the time to treatment for acute myocardial infarction. AB - PURPOSE: To compare delay and circumstances of decisions to seek care in patients with acute myocardial infarction (AMI) in the United States (US), England, Australia, South Korea, and Japan. DESIGN: Comparative prospective design. METHODS: Patients diagnosed with AMI (N = 913) were interviewed within 72 hours of hospital admission for confirmed AMI using the Response to Symptoms Questionnaire. Delay times were calculated from review of emergency room records and patients' interviews. Analysis of variance was used to test differences in delay time among countries. FINDINGS: Median delay ranged from 2.5 hours in England to 6.4 hours in Australia, with the three Pacific Rim countries reporting median delay times > 4 hours. The majority of patients experienced initial symptoms at home (range: 56% in Japan to 73% in the US) with the most common witness being a family member (32% in South Korea to 48% in England). Ambulance use was widely divergent with the highest use in England (85%) and the lowest use in the US (42%). CONCLUSIONS: In all countries, median delay was too long to obtain maximum benefit from AMI therapies, particularly thrombolysis. Education and counseling of patients and families to reduce prehospital delay in AMI episodes might be more effective if the various factors influencing patients' first responses to symptoms are considered, as well as differences in health care systems. PMID- 14735674 TI - Social capital, health, and health disparities. AB - PURPOSE: To synthesize the empirical evidence that links social capital to population health with the aim of identifying implications for health disparities research. METHODS: A literature search of PubMed and CINAHL databases from January 1990 to June 2002 was done using the search term "social capital." In addition, tables of contents of applicable journals from January 1997 to June 2002 were searched. Reference lists were examined for additional empirical and theoretical articles related to social capital and health. Eighty-four articles were retrieved for review and 19 articles met inclusion criteria. FINDINGS: Although most reseachers concluded that their findings supported an association between social capital and health, all research was descriptive, without conceptual development. This gap resulted in (a) lack of distinction of the concept as an attribute of a geographic space or as an individual attribute, (b) problematic use of operational variables, and (c) limited theoretical exploration of causal linkage. These deficits limit the usefulness of the concept for health disparities research. CONCLUSIONS: The lack of conceptual development diminishes the usefulness of social capital as a variable for public health research. However, the empirical evidence is sufficient to warrant further work to advance the concept in relation to population health and health disparities. PMID- 14735675 TI - Testing an intervention for preventing osteoporosis in postmenopausal breast cancer survivors. AB - PURPOSE: To test a 12-month multicomponent intervention for preventing or treating osteoporosis in 21 postmenopausal women who had completed treatment (except Tamoxifen) for breast cancer, and for whom hormone replacement therapy (HRT) was contraindicated. DESIGN: Pilot intervention study. METHODS: The intervention consisted of home-based strength and weight training exercises, 5 or 10 mg alendronate per day, 1500 mg calcium per day, 400 IU vitamin D per day, education on osteoporosis, and facilitative strategies to promote adherence to the intervention. Outcome measures were: adherence to the intervention, dynamic balance, muscle strength, and bone mineral density (BMD) of the hip, spine, and forearm. FINDINGS AND CONCLUSIONS: Adherence to calcium, vitamin D, and alendronate therapy was above 95%, and adherence to strength training exercises was above 85%. Over the 12 months, the 21 participants had significant improvements in dynamic balance, muscle strength for hip flexion, hip extension, and knee flexion, and BMD of the spine and hip. Participants had a significant decrease in BMD of the forearm. Three of the 21 women who had measurable bone loss at baseline had normal BMD after 12 months of the intervention. PMID- 14735676 TI - Contrasts in patients' and providers' explanations of rheumatoid arthritis. AB - PURPOSE: To investigate explanations of rheumatoid arthritis (RA) from young women's perceptions of the illness experience and providers' understanding of the disease. DESIGN AND METHODS: This ethnographic study included 17 women from age 26 to 40 years who were under medical care for RA, and five health care providers of these participants. FINDINGS: Two main themes were identified: (a) having RA "is a pain" and (b) it changed me. The first theme represented the participants' physical and emotional suffering, interferences in their everyday lives, and aggravations in receiving health care. The second theme represented the life and lifestyle changes the women experienced as a result of having RA. The providers' explanations included two themes: (a) functioning in a normal manner and (b) controlling the disease. The first theme was keeping the women's physical functioning as normal as possible. The second theme was the providers' goal to control the disease, which they believed would lead to the preservation of joint function and manageable pain levels. Regarding results of health care, the participants and providers had comparable beliefs about decreasing the physical pain and improving joint function but they had disparate notions about the participants' being active partners in communication and negotiation processes of their health care. CONCLUSIONS: The findings indicated the importance of discovering potential disparities in patients' and providers' explanations of RA, and revealed the participants' desire for clinical support in becoming partners in their own health care. PMID- 14735677 TI - Birth outcomes and maternal morbidity in abused pregnant women with public versus private health insurance. AB - PURPOSE: To compare the effects of recent intimate partner abuse on maternal and infant health in publicly versus privately insured pregnant women. DESIGN: Exploratory descriptive analysis in 13 Massachusetts prenatal care sites from records of 2,052 women who had been screened during pregnancy for domestic violence. METHODS: Clinicians screened pregnant women for domestic violence using the Abuse Assessment Screen. After delivery, prenatal and birth outcome data and abuse screening results were extracted from medical records by project staff. Odds ratios were used to compare maternal and infant health indicators in abused and nonabused women. Data from women with public and private health insurance then were examined separately, using logistic regression to control for low education and single marital status while examining the odds of adverse maternal and infant outcomes in abused and nonabused women. FINDINGS: In the sample as a whole, recently abused women were more likely to be publicly insured and unmarried, to have less than 12 years of formal education, and to have medical and obstetrical complications. Parity, ethnic background, and infant birth outcomes did not differ in relation to abuse. In separate analyses for women with public and private health insurance, after controlling for marital status and education, abuse increased the odds of low infant Apgar scores, poor nutrition, hyperemesis, hypertension, and substance abuse in publicly insured women, and abuse increased the odds of poor nutrition and bleeding during pregnancy for privately insured women. CONCLUSIONS: The different correlates of abuse in publicly and privately insured women might be important for clinicians caring for these different populations. Screening for abuse and providing abuse-related services are indicated for pregnant women. PMID- 14735678 TI - American and Icelandic parents' perceptions of the health status of their young children with chronic asthma. AB - PURPOSE: To identify factors that influence American and Icelandic parents' health perceptions among families of infants or young children with asthma. DESIGN: A cross-sectional research design of 76 American families and 103 Icelandic families. Data were collected mainly in the Midwest of the United States (US) and in Iceland from August 1996 through January 2000. METHOD: Parents in these two countries who had children aged 6 or younger with chronic asthma completed questionnaires regarding family demands, caregiving demands, family hardiness, sense of coherence, and health perceptions. Descriptive statistics, chi-square tests and t tests were compiled. Multiple regression analysis was used to test path models and for mediation. FINDINGS: American parents differed from their Icelandic counterparts in family hardiness. In both countries, significant differences were found in caregiving demands and health perceptions between mothers and fathers. Illness severity and caregiving demands affected health perceptions of both mothers and fathers. Sense of coherence mediated the relationship between family demands and parents' perceptions for both parents. For mothers only, family hardiness mediated the relationship between family demands and health perceptions. CONCLUSIONS: The Resiliency Model of Family Stress, Adjustment, and Adaptation was useful for building knowledge on parents' health perceptions in two Western cultures for families of young children with asthma. Interventions emphasizing family and individual resiliency and strengths have the potential to affect parents' views of their children's health. PMID- 14735679 TI - Challenging the myths about parents' adjustment after the sudden, violent death of a child. AB - PURPOSE: To examine three commonly held myths: (a) a child's death by suicide results in the worst parental outcomes compared with other causes of violent death, (b) divorce is not only more common among bereaved than nonbereaved married couples, it might be inevitable, and (c) "letting go and moving on" is an essential bereavement task needed for a satisfactory adjustment following the violent death of a child. DESIGN AND METHODS: Review of empirical evidence and critical reviews, review of Internet resources available to the general public, and the inclusion of original data obtained from a longitudinal, prospective study conducted by the authors. FINDINGS: Conclusive evidence was found to dispel two of the three myths, but sufficient evidence was not found to draw conclusions about the third myth regarding parents' adjustment to a child's suicidal death. CONCLUSIONS: Myths in regard to parental bereavement are resistant to disconfirming evidence and they appear to persist among professional practitioners and the general public despite contrary empirical evidence. PMID- 14735680 TI - Cancer patients' demographic characteristics and ratings of patient-centered nursing care. AB - PURPOSE: To examine relationships between patients' demographic characteristics and patients' reports of patient-centered care. DESIGN: Secondary analysis of data (N = 423) from a study in the northeastern United States focused on the psychometric properties of the Oncology Patients' Perceptions of the Quality of Nursing Care Scale (OPPQNCS). METHODS: The quality of four interpersonal nursing interventions, representing patient-centered nursing care, was measured with the OPPQNCS subscales. Patients' characteristics included race (White or non-White), sex, age, education, income, and hospitalization for cancer. Four separate ordinary least squares regression models were constructed. FINDINGS: Hospitalization was inversely related to intervention quality in each model. Income was the only statistically significant characteristic for nonhospitalized patients, and only in the coordination model. For hospitalized patients, education was statistically significant in the coordination model, and income in the proficiency model. An interaction term for education and income was statistically significant in the responsiveness model, and a term for gender and education in the individualization model. CONCLUSIONS: Hospitalized patients' exposure to nursing care may indicate a wider range of care quality than for nonhospitalized patients, possibly accounting for the inverse relationship between hospitalization and patient-centered care. Groups identified at risk for lower quality care--minorities, women, elders, and people in low-income groups- did not report a lower level of patient-centered nursing care. Nurses' contributions to patient-centered care and care equity are important components of care quality, particularly for hospitalized cancer patients. PMID- 14735681 TI - Issues and dilemmas in conducting research with vulnerable home hospice participants. AB - PURPOSE: To describe issues and dilemmas related to nonparticipation, attrition, and needs for assistance in research with vulnerable home hospice participants. DESIGN AND METHODS: Retrospective analysis, with descriptive statistics of the frequency of issues and dilemmas that occurred in a research study with a vulnerable population. FINDINGS: From a group of 113 potential participants, 16 (14.1%) people who gave initial consent were unable to participate or were lost to the study (subset I) for the following reasons: unable to give informed consent, cognitive disturbance, and physical distress. Of the 97 participants who completed testing, 28 (28.8%) required assistance (subset II) because of poor vision, physical weakness, and other physical impairments. CONCLUSIONS: Loss of potential participants limited this study and hampered effect size. Research with home hospice patients required careful assessment for symptoms that precluded informed consent. Issues with cognition indicated need for a tool to assess mental acuity. Although several participants required assistance, those who completed testing expressed gratitude at being able to contribute information that they believed would benefit others. PMID- 14735682 TI - Understanding and responding to patients' requests for assistance in dying. AB - PURPOSE: To explore how nurses experience and respond to patients' requests for assistance in dying (AID). DESIGN AND METHODS: A phenomenological study of 10 self-selected nurses. FINDINGS: Four major themes: Being Open to Hear and Hearing; Interpreting and Responding to the Meaning; Responding to Persistent Requests for AID, and Reflections. When faced with persistent requests for AID, participants provided a continuum of interventions: refusal, providing palliative care that might secondarily hasten dying, respecting and not interfering with patients' or families' plans to hasten dying, and providing varying types and degrees of direct AID. Their responses were context-driven rather than rule mandated, and they drew a distinction between secondarily hastening and directly causing death. CONCLUSIONS: Few nurses in this study unequivocally agreed or refused to directly help a patient die. Most struggled alone and in silence to find a morally and legally acceptable way to help patients who persisted in requesting AID. Regardless of how they responded, many described feelings of conflict, guilt, and moral distress. PMID- 14735683 TI - Home care nurses' descriptions of important agency attributes. AB - PURPOSE: To identify attributes of home care agencies described by nurses as important to their professional practice and job satisfaction. DESIGN: Seven focus groups with home health care staff nurses were conducted at 6 home care agencies located in three states in the Mid-Atlantic region of the United States. A total of 58 home care nurses participated in the study. METHODS: Transcriptions of tape-recorded focus groups were subjected to open and axial coding techniques. RESULTS: Six major categories and eight subcategories of organizational attributes described by home care nurses as important to the support of their practice and job satisfaction were identified. CONCLUSIONS: Attributes described by home health nurses were not only similar to those described by hospital-based nurses as reported in the "magnet hospital" literature, but they also were consistent with key concepts described in theories of professional workforce organization. Findings provide insight into developing workplace environments to support home care nurses. PMID- 14735684 TI - Experiences of pioneer nurse practitioners in establishing advanced practice roles. AB - PURPOSE: To describe pioneers' experience of establishing the nurse practitioner (NP) role, and their experiences in maintaining and building the NP role in the contemporary practice environment. DESIGN: The study sample included 50 middle aged women currently practicing in Washington State as licensed advanced practice nurses, who began the NP role during 1965-1979. METHODS: This descriptive study included interviews and focus groups to gather data about the nurses' early experiences. Interpretative methods of grounded theory were used in data collection and analysis. FINDINGS: The central organizing theme, Advancing Autonomy to Make a Difference, was manifested through six broad themes: Breaking Free, Molding the Clay, Encountering Obstacles, Surviving the Proving Ground, Staying Committed, and Building the Eldership. CONCLUSIONS: Autonomy was requisite to practice to one's full potential and maintain commitment over time. The data findings show the evolution of advanced nursing practice in the United States and provide guidance for nurses who are working to establish advanced practice nursing in other countries. PMID- 14735685 TI - Health-related quality of life in patients with advanced metastatic melanoma: results of a randomized phase III study comparing temozolomide with dacarbazine. AB - Health-related quality of life (HRQL) is a crucial endpoint in the evaluation of treatments that have limited survival benefits. The HRQL evaluations help ensure that patients are not sacrificing life quality for quantity. Current treatments for metastatic melanoma are primarily palliative, because cure is unattainable. The purpose of this article is to report detailed HRQL results of a phase III clinical trial comparing temozolomide to dacarbazine (DTIC) in patients with metastatic melanoma. Patients were randomized to receive either oral temozolomide for 5 days every 4 weeks or intravenous DTIC for 5 days every 3 weeks. The HRQL was evaluated on day 1 cycle 1 and after each subsequent treatment cycle using the EORTC QLQ-C-30. The HRQL was compared between groups at weeks 12 and 24. Patients treated with temozolomide reported significantly better physical functioning and less fatigue and sleep disturbances than patients treated with DTIC at week 12. For all but two function and symptom subscales, EORTC QLQ-C30 subscale scores were numerically better for patients treated with temozolomide at week 12. All subscales except diarrhea were better for temozolomide at week 24. Analyses of change scores revealed that patients treated with temozolomide reported statistically significant improvements in emotional well-being and sleep disturbance. Patients also reported near significant change in cognitive functioning (3.9, p = 0.06). Patients treated with DTIC deteriorated on most function subscales and many symptom subscales at week 12. Deterioration in physical functioning approached significance (-6.8, p = 0.06). At week 24, patients treated with DTIC improved on the emotional functioning subscale and deteriorated on the physical, role, and global HRQL subscales, although many of the symptom scores improved. The results of this study suggest that treatment with temozolomide leads to important functional improvements and decreased symptoms compared to treatment with DTIC in patients being treated for metastatic melanoma. PMID- 14735686 TI - Alternating doublets: establishing the optimal multifractionated dosing schedule to administer docetaxel, cisplatin, gemcitabine, and vinorelbine in combination. AB - The purpose was to determine the optimal multifractionated (MF) dosing schedule to permit the delivery of four active agents in nonsmall cell lung cancer simultaneously in alternating doublets (docetaxel-cisplatin alternating with gemcitabine-vinorelbine). Three MF schedules were used: schedule A weekly; schedule B twice weekly for 2 weeks repeated every 21 days; and schedule C twice weekly every other week. Dose fractions were fixed for each component drug: docetaxel 50 mg/m2 plus cisplatin 20 mg/m2 and gemcitabine 500 mg/m2 plus vinorelbine 25 mg/m2. GCSF 480 micrograms was administered as a single dose concomitant with chemotherapy if the WBC was between 1500 and 3500 cells/mm3. Hematological toxicity, particularly leukopenia and anemia, was the predominant adverse effect observed and was demonstrated on all three schedules. Schedule B was not feasible in that none of the seven cycles were completed and six of seven required hospitalization for febrile neutropenia. The delivery of a four-drug combination chemotherapy regimen consisting of docetaxel, cisplatin, gemcitabine, and vinorelbine is feasible with an alternating doublet multifractionated dosing scheme with either a weekly or twice weekly every other week schedule. PMID- 14735687 TI - A dose-escalating pilot study of sterically stabilized, pegylated liposomal doxorubicin (Lipo-Dox) in patients with metastatic breast cancer. AB - Doxorubicin is considered among the most active single agents used against advanced breast cancer. Recent advances in the design of liposomes as carriers of cytotoxic drugs have resulted in a new formulation of doxorubicin with improved pharmacokinetic and tumor-localizing properties. The objectives of this dose escalating pilot study were to evaluate the efficacy and safety of the sterically stabilized, pegylated liposomal doxorubicin (Lipo-Dox) for the treatment of metastatic breast cancer. Lipo-Dox was given at the dosage of 45 mg/m2 over 1 hr of intravenous infusion every 4 weeks initially and could be escalated up to a maximum of 60 mg/m2. Response was assessable in 17 of 19 intent-to-treat patients. An objective response was achieved in 41.2% (95% confidence interval: 17.8%-64.6%) of patients (5.9% complete response and 35.3% partial response), and 23.5% had stable disease. Median time to disease progression was 163 days. Major treatment-related toxicities included neutropenia, stomatitis, and skin toxicity in this dose-escalation program. Impressively, no grade 4 toxicities have ever been observed. The only grade 3 nonhematological toxicity ever to occur was reversible skin toxicity, presented as palmar-plantar erythrodysthesia. No severe nausea/vomiting, wig-necessary alopecia, or significant cardiac function change were encountered. In conclusion, Lipo-Dox is shown by this first reported pilot study to be an active agent for treatment of advanced breast cancer with a safety profile that differs markedly from free doxorubicin. The dosage of 45-60 mg/m2 every 4 weeks was well tolerated. Because myelosuppression and other nonhematological toxicities associated with Lipo-Dox were generally mild and acceptable, further assessment of this drug particularly in combination with other chemotherapeutic drugs in the management of early or advanced breast cancer is suggested. PMID- 14735688 TI - Arterial desaturation syndrome following pleurodesis with talc slurry: incidence, clinical features, and outcome. AB - The objectives were to define the incidence, risk factors, clinical features and outcome of arterial desaturation syndrome following talc pleurodesis in patients with malignant pleural effusions. This retrospective, observational study took place at a tertiary care cancer center in New York. All patients were those with malignancy who underwent pleurodesis with talc in 1998 at Memorial Sloan Kettering Cancer Center. Characteristics of patients are described by using summary statistics. Differences between groups were assessed with the Fisher's exact statistic for categorical variables and Student's t-test for continuous variables. Among patients who were considered to have arterial desaturation syndrome, we evaluated the relation of SaO2/FIO2 pre- and post-talc installation using a paired Student's t-test. During 1998, 120 patients underwent pleurodesis with talc, and 8 (7%) developed arterial desaturation following the procedure. Symptoms included chest pain, dyspnea, fever, and increased need for oxygen supplementation developed typically within 1 day. Three of the eight patients in this series required mechanical ventilation, but all recovered uneventfully after treatment, which included high-dose corticosteroids. Patients with breast and ovarian cancer appeared to be at increased risk for this complication compared to those patients with other types of cancer (p = 0.01). Approximately 7% of patients who have undergone sclerosis with talc for a malignant pleural effusion will develop arterial desaturation with clinically significant hypoxia requiring supplemental oxygen following the procedure. It appears that most patients recover from this complication and that those with breast and ovarian cancer may be at higher risk. PMID- 14735689 TI - A phase II study with CPT-11 plus leucovorin and bolus IV 5-fluorouracil in patients with advanced colorectal carcinoma. AB - Standard chemotherapy in advanced colorectal carcinoma (CRC) has not yet been established. The present study was conducted to assess the efficacy and toxicity profile of CPT-11, leucovorin (LV), and bolus 5-fluorouracil (5-FU) in a weekly schedule. Fifty-five patients were entered with no prior chemotherapy for advanced disease or adjuvant treatment ended at least 6 months preceding study entry, and 45 were assessable for response. Patients were treated with CPT-11 80 mg/m2 (7 patients) or 70 mg/m2 (48 patients). After completion of CPT-11 infusion, LV 200 mg/m2 was administered over 2 hr followed immediately by 5-FU 450 mg/m2, IV bolus, weekly for 6 weeks followed by a 2-week rest period. Treatment was continued for four cycles. Because of grade 3 and 4 diarrhea in four of the first seven patients, the study was amended to reduce the starting dose of CPT-11 from 80 to 70 mg/m2 weekly. Four complete and 10 partial responses were observed (response rate: 25.5%), the median time to progression (TTP) was 7.7 months, 1-year survival rate was 62.3%, and the median overall survival was 15.0 months. Grade 3 and 4 diarrhea occurred in seven patients (12.7%), four of them treated with CPT-11 80 mg/m2. Grade 3 myelotoxicity occurred in five patients (9.0%). Toxic death because of diarrhea, neutropenia, bacteremia, and sepsis occurred in a patient treated with CPT-11 80 mg/m2. Our results confirm the efficacy of CPT-11, LV, and 5-FU in a weekly schedule in patients with advanced CRC. Further studies are needed to compare the present regimen with higher doses of CPT-11 with LV plus different schedules of 5-FU administration in the treatment of metastatic CRC. PMID- 14735690 TI - Usefulness of thallium-201 SPECT scintimammography to differentiate benign from malignant breast masses in mammographically dense breasts. AB - The purpose of this study was to assess the usefulness of thallium-201 (Tl-201) single-photon emission-computed tomography (SPECT) scintimammography to differentiate benign from malignant breast masses in 32 female Taiwanese patients with indeterminate mammographic probability of malignancy because of mammographically dense breasts. All breast masses were removed, and final histopathological diagnoses were obtained in all cases. The results showed that thallium-201 SPECT scintimammography findings were true-positive in 22 cases, false-positive in 1 case, true-negative in 7 cases, and false-negative in 2 cases. The diagnostic sensitivity, specificity, and accuracy were 91.7%, 87.5%, and 90.6%, respectively, for detecting breast cancer in mammographically dense breasts. In conclusion, thallium-201 SPECT scintimammography is a useful tool for differentiating benign from malignant breast masses in patients with indeterminate mammograms because of mammographically dense breasts. PMID- 14735691 TI - Slowing growth and histology changes in Lewis lung carcinoma implanted in a partly denervated muscle. AB - The purpose of this study is to test the effect of partial muscle denervation on a tumor inocculated on the same muscle. Experiments were performed on male C57BL mice divided into two numerically equal A and B groups, each of which was subdivided into three subgroups AA, AB, Ac and BA, BB, BC, respectively. The right sciatic nerve of each mice of the A group was incised and partly dissected (day 0). Then Lewis lung carcinoma (LLC) cells were implanted on the right gastrocnemia of each mice of both groups on day 0 for subgroups AA and BA, on day 1 for subgroups AB and BB, and on day 4 for subgroups AC and BC. The incision and dissection of the sciatic nerve caused a significant reduction in the size of the tumor in all three A subgroups of mice. Fourteen days after implantation, the size of the primary tumors implanted on the right hind leg was 2.6 +/- 0.32 cm3, 3.5 +/- 0.21 cm3, and 4.7 +/- 0.31 cm3 for the three BA, BB, BC subgroups of the control group compared to 1.4 +/- 0.19 cm3, 2.0 +/- 0.28 cm3, and 3.0 +/- 0.27 cm3 for the three subgroups AA, AB, Ac, in which sciatic nerve was incised and dissected, indicating inhibition of the tumor growth of 46.2%, 43.0% and 36.2%, respectively. Although a smaller part of gastrocnemius is innervated by the anterior femoral nerve, which was left intact, denervation of the sciatic nerve caused inhibition of the tumor growth. This may be due to worse nutrition and/or blood supply of the LLC tumor. Mitochondria function and the degree of apoptosis of the tumor cells are further studied by us. PMID- 14735692 TI - The first 1000 dendritic cell vaccinees. AB - Dendritic cells (DCs) are potent antigen-presenting cells that have the ability to stimulate primary T cell antitumor immune responses in animals and humans. Since the first published clinical trial of dendritic cell vaccination in 1995, 98 studies describing more than 1000 vaccinees have been published in peer reviewed medical journals or presented at the annual meetings of the American Society for Clinical Oncology, the American Association of Cancer Research, or the American Society of Hematology. Trials have been performed in 15 countries. Trials included patients with more than two dozen tumor types; most trials studied patients with malignant melanoma, prostate cancer, colorectal carcinoma, or multiple myeloma, using autologous DCs pulsed with synthetic antigens or idiotype antibodies. The DC vaccines were also prepared by pulsing DCs with tumor lysates or RNA, by transfection with tumor DNA, or by creating tumor cell/DC fusions. Various approaches to vaccine cell numbers, length of vaccine program, site of vaccination, frozen preservation of vaccine, and use of a maturations step for DCs were used. Adverse effects associated with DC vaccination were uncommon; most were mild and self-limited and none were serious. Clinical responses were observed in approximately half the trials. The DC vaccination may provide a safe approach to cancer immunotherapy that can overcome the limited reach and immunogenicity of peptide vaccines. PMID- 14735693 TI - Paclitaxel and concurrent radiation in upper gastrointestinal cancers. AB - Effective locoregional treatments are needed for adenocarcinomas of the esophagus, stomach, and pancreas. Paclitaxel has been investigated as a radiation sensitizer for upper gastrointestinal malignancies. In esophageal cancer, the combination of low-dose weekly paclitaxel, platinum, and concurrent radiation therapy (RT) has substantial activity and is well tolerated. Regimens that add fluorouracil (5-FU) to paclitaxel and platinum or incorporate hyperfractionation radiation have a higher incidence of severe esophagitis. In gastric cancer, adjuvant concurrent paclitaxel, 5-FU, and radiation is being investigated in the cooperative group setting. In pancreatic cancer, paclitaxel may be a radiation sensitizer even to tumor cells that are resistant to paclitaxel as a single agent. The Radiation Therapy Oncology Group (RTOG) demonstrated a 43% 1-year survival with paclitaxel/RT for patients with locally advanced pancreatic cancer. This represented a 40% improvement in survival compared to the previous RTOG 92 09 study of 5-FU-based chemoradiation. Ongoing trials in pancreatic cancer are investigating the addition of gemcitabine to paclitaxel and radiation and incorporating molecular targeting agents. PMID- 14735694 TI - Genetics of neurofibromatosis 1-associated peripheral nerve sheath tumors. AB - Neurofibromatosis 1, an inherited disorder that affects 1/3500 individuals worldwide, predisposes to the development of benign and malignant peripheral nerve sheath tumors. The disorder results from inactivation of one of the NFI genes. The second NFI gene is typically inactivated in Schwann cells during tumor formation. This article reviews the different types of genetic alterations in NFI in both constitutional and tumor tissues and genetic alterations of other genes that may affect tumorigenesis. These studies have provided insight into the genetic basis of both the variable expression of the disorder and of benign and malignant peripheral nerve sheath tumorigenesis. PMID- 14735695 TI - Family-oncologist communication in cancer patient care. PMID- 14735696 TI - Bryostatin-1: a novel PKC inhibitor in clinical development. AB - Modulation of PKC represents a novel approach to cancer therapy. Bryostatin-1 is a macrocyclic lactone derived from a marine invertebrate that binds to the regulatory domain of protein kinase C. Short-term exposure to bryostatin-1 promotes activation of PKC, whereas prolonged exposure promotes significant downregulation of PKC. In numerous hematological and solid tumor cell lines, bryostatin-1 inhibits proliferation, induces differentiation, and promotes apoptosis. Furthermore, preclinical studies indicate that bryostatin-1 potently enhances the effect of chemotherapy. In many cases, this effect is sequence specific. Bryostatin-1 is currently in phase I and phase II clinical trials. The major toxicities are myalgias, nausea, and vomiting. Although there is minimal single-agent activity, combinations with standard chemotherapy are providing very encouraging results and indicate a new direction in cancer therapy. PMID- 14735697 TI - Looking beyond morphology: cancer gene expression profiling using DNA microarrays. AB - Although molecular testing has been increasingly used in clinical practice, the precise diagnosis and prognosis of most human cancers still heavily rely on descriptive histopathological data. Identification of robust molecular markers associated with distinctive morphological parameters will assist in diagnostic and prognostic assessment. On the other hand, the diversity of morphologically identical/similar cancers can be manifested at multiple levels, most importantly at the level of clinical outcome. In the past several years, DNA microarray technology has been widely used in the context of cancer research, resulting in a deluge of new information that can be used to identify molecular alterations common to all tumors as well as signatory profiles unique to a subcategory of cancers. These new findings will very likely transform the clinical management of cancer patients in the near term. This article reviews recent advances in cancer gene expression-profiling results derived from the application of DNA microarray technology, with an emphasis on studies performed on human prostate cancer specimens. We discuss broad issues relevant to cancer expression profiling and attempt to illustrate the rapid pace of novel discoveries using prostate cancer as examples wherever appropriate. PMID- 14735698 TI - Applications of image analysis to anatomic pathology: realities and promises. AB - Image Analysis in Pathology is viewed as an ancillary method meant to provide objective support in the resolution of difficult problems. Its Achilles heel is the process of nuclear segmentation (delimitation of the nuclear membrane) which is extremely difficult in pathology materials. Although interactive segmentation procedures are available no reliable fully automatic method has been described. The only application of image analysis that has truly succeeded in Pathology is DNA ploidy measurement. A very desirable application is the quantitation of immunohistochemical markers, which is technically challenging, has been resolved only in certain cases and is unlikely to have a general solution. Nuclear quantitation has repeatedly proven to be helpful in reaching differential diagnoses, in particular when based on size distributions of nuclear profiles rather than its average, but is hampered by the segmentation problem discussed above. Texture analysis of chromatin is an exciting, mathematically complex application likely to succeed, for which many approaches have been described. Finally a diagnosis (classification) can be obtained based on algorithms applied to multiple descriptors of tumor cells (for instance nuclear sizes, chromatin texture, shape, etc). The best classificatory approaches are neural networks (a form of artificial intelligencee), multivariate analysis, and logistic regression (statistical). PMID- 14735699 TI - Prevent and relieve suffering: professional palliative care. PMID- 14735700 TI - Quality of life, at what cost? PMID- 14735701 TI - Doxorubicin for metastatic breast cancer: time for a change? PMID- 14735702 TI - Dendritic cell vaccines--hopeful or hopeless? PMID- 14735703 TI - We're only partway there. HHS' first annual quality report shows some big strides have been made but points out acute need for more preventive care. AB - HHS has delivered its first report card on healthcare quality, with results that showed progress in the treatment of cancer, diabetes, end-stage renal disease and heart attacks, but pointed out some areas for big improvement, like preventive care. The report will help hospitals in their goal of giving patients the right care, says Richard Davidson, left, president of the American Hospital Association. PMID- 14735704 TI - States of frustration. Hospital associations, desperate over shortfalls, are resorting to hospital taxes. PMID- 14735705 TI - Back to basics. AMA simplifies philosophy on membership. PMID- 14735706 TI - A primary concern. Medicare provision ensures program is second payer; $9 billion savings seen. PMID- 14735707 TI - Necessary knowledge. Senators craft bill to create federal nurse database accessible to all employers. PMID- 14735708 TI - Shuttling toward a safety culture. Healthcare can learn from probe panel's findings on the Columbia disaster. PMID- 14735709 TI - Racing toward market mania. Healthcare industry gearing up for big changes in 2004, starting with policy at state level and adjusting to Medicare reform law. PMID- 14735710 TI - Making the rounds. Stanford's Martha Marsh says she believes in getting to know hospitals she runs from bottom up. PMID- 14735711 TI - [1995, women's year]. PMID- 14735712 TI - [Senescence process of mnesic centers: a study in humans and rats]. AB - The age-related neuronal changes in mnemonic centres were studied in 37 human and 36 rats (Wistar) brains. The age of the brains (without cerebral alterations) was uniformly distributed along the lifespan (16-86 years in humans and 1-36 months in rats). The results showed parallel changes in humans and rats. The neuronal loss oscillated between 3 and 64%, mean 32%. Neuronal death was a continuous process, although there were differences according the centres, for instance, the higher loss was found in the first half of life in the cortex entorhinalis and hippocampus (rat), and the contrary happened in the dorsolateral and basomedial nucleus of amygdala. In other centres, e.g., mamilla body, basal nucleus of Meynert etc. the loss was quite uniform. The modification in nuclear size showed 3 different phases: there was an initial period in which the nuclear area decreased, a second period with an increase and, in the last period, there was a stabilization in humans and a conspicuous decrease in rats. The nuclear enlargement is interpreted as result of the loss of redundance in nervous centres and the stabilization or atrophy as a consequence of loss of the neuronal plasticity. PMID- 14735713 TI - [Extra-abdominal desmoid tumors. Diagnostic-therapeutic considerations and review of the literature]. AB - The authors review the literature, discuss clinical aspects and options in the management of desmoid tumors, analyze recent advances in the understanding of its biology and how they affect therapeutic choices, and outline future treatment perspectives. We conclude that although recurrence rates are high, wide excision is the best firs approach. If it is not possible, marginal surgery and radiation therapy can be performed. Radiation therapy can be also use in the attempt to control recurrent disease. Although many pharmacologic agents have been advocated there is no clear proof of their usefulness. Systemic therapy may be considered for selected patients that are not amenable to local treatment. PMID- 14735714 TI - [Study of placenta blood flow throughout pregnancy]. AB - The human placenta has a double vascularization which provides with two different blood flows: the uteroplacental and fetoplacental or umbilical ones. Gestational success depends on its adequate interrelationship. Uteroplacental blood flow experiences a progressive increase during gestation, being at term 17 to 20-fold that of non-pregnant uterus (600 ml/min). There are two periods of greater increase: placentation and the last part of gestation. Umbilical blood flow increases in parallel with fetal weight. Uteroplacental and umbilical blood flows are influenced by nervous, chemical and mechanical factors, but the main determinant are the vascular changes which include histological and growing and development phenomena. PMID- 14735715 TI - [Tacrolimus (FK 506): immunosuppressive alternative to solid organ transplant (Part 1)]. PMID- 14735716 TI - [Ketorolac]. PMID- 14735718 TI - [The brain decade (1990-2000)]. PMID- 14735717 TI - [Palliative management of patients with advanced AIDS]. AB - Patients with AIDS in terminal stages may improve with diverse technics and philosophy of terminal care according to their special features. We review the clinical course of the illness produced by HIV infection, the most bothering symptoms in this stage, the preventive measures and treatment. The patient and his family need a strong support to endure this disease. PMID- 14735719 TI - [Production and characterization of a monoclonal antibody that selectively marks the astrocyte population in the central nervous system]. AB - The development of a monoclonal antibody that selectively labels the astrocitic population of the central nervous system in the three species tested (human, monkey and rat) is described. The tissue used as immunogen consisted in a sector of the human hippocampus (CA1) from a brain removed shortly after death. The neuropathological examination in this case revealed an congophilic angiopathy. The tissue was frozen until homogenezation and intraperitoneal injection in 8 weeks old BALB/C mice, after which lymphocytes from the spleen were fused with NS 1 myeloma cells devoid of the enzyme Hypoxantine Guanine Phosphoribosyl Transferase (HPGT (-)) following standard procedures. The resulting hybridomas were tested by ELISA assays, immunofluorescence and immunohistochemical (PAP and ABC) techniques. The monoclonal antibody obtained was identified as an IgM and the Western blot analysis indicated that it recognizes a protein of 43,000 dalton. Immunohistochemistry was performed in 30 or 50 microns sections of human temporal lobe perfused through the carotid arteries with a mixture of 4% paraformaldehyde and 0.02% picric acid. Monkey tissue (sectioned at 50 microns) was obtained from a brain with the same fixation. Rat tissue followed the same protocol, although better results were seen after shifting to 1% paraformaldehyde and 1.25% glutaraldehyde (sections were 30 microns thick). Paraffin embedded human tissue (fixed in either 10% buffered formalin or in 4% paraformaldehyde) was also tested. All immunoreactions were intensified in a solution of osmium tetroxide and thiocarbohydrazide. The monoclonal antibody gave best results at a dilution of 1:50-100, where it gave a sharp labeling of both fibrous and protoplasmic astrocytes up to the most delicate processes, both in white and the grey matter. Rat tissue fixed in only 4% paraformaldehyde resulted in less optimal labeling, mostly restricted to limiting glia and Bergmann glia. The other fixation protocol (1% paraformaldehyde and 1.25% glutaraldehyde) gave a labeling comparable to monkey and human tissue. Paraffin embedded tissue was positive and similar to that obtained in frozen tissue, only when used the 4% paraformaldehyde fixation, but not when it was fixed in 10% formalin. Therefore, this monoclonal antibody has high specificity for the astrocitic population in the nervous system of mammals and it can be useful in a variety of studies of the normal and pathological situations of the nervous system. PMID- 14735720 TI - [Assessment of the hemostatic system in healthy preterm and fullterm newborns]. AB - Assessment of the hemostatic system in premature and newborn infants. When the newborn is compared to older children and adults, several differences in the coagulation and fibrinolytic systems have been described; near-adult values are achieved for most components by 6 months of life. The newborn has an impaired platelet aggregation, reduced synthesis of clotting factors and inhibitors and molecular abnormalities in some proteins of blood coagulation. The role of fibrinolysis is, however, less well known. We have performed a study to assess the fibrinolytic activity in 100 healthy newborns and we have found a marked increase of the fibrinolytic activity, mainly related to an enhancement of plasminogen activators (t-PA) and reduction of fibrinolysis inhibitors, without systemic fibrinolysis. These results would suggest that the fibrinolytic system in the newborn is in equilibrium with the clotting system in order to preserve physiological hemostasis. PMID- 14735721 TI - [Filgrastim: granulocyte colony stimulating factor]. PMID- 14735722 TI - [Histiocytosis X]. AB - We make a clinical and bibliographical review in diagnosis and treatment of Histiocytosis X, who includes three components: Eosinophylic granuloma, Hand Schuller-Christian disease and Letterer-Siwe syndrome. We had 28 cases, with an average age of 11.8 years. Eighteen (64%) were males and 10 (36%) females. Seventeen were solitary lesions and 8 had multiple skeletal lesions. Three patients had generalized disease: 2 with Hand-Schuller-Christian disease and 1 with Letterer-Siwe syndrome. The femur lesion was frequently observed among the solitary lesions and the cranium involvement in the multiple skeletal lesions. The total number of lesions was 47. The treatment depended on the location, the clinical manifestations and the number of lesions. The patients with solitary lesions (17) were treated with curettage and bone grafting in 6 cases, segmental resection in 3 cases, only curettage in 3, radiotherapy in one and 4 patients didn't receive treatment. The patients with multiple lesions (8), were treated surgically (4 cases), only radiation therapy (2 patients), with surgery and radiotherapy (1) and one patient had no treatment. The generalized disease was treated with surgery of the main lesion depending in the clinical manifestations and chemotherapy and radiotherapy. The prognosis was good in almost all the patients and we observed regression of the disease even without specific treatment. The case of acute generalized disease died due to systemic complications. PMID- 14735723 TI - Advances in the pathogenesis of liver cirrhosis. PMID- 14735724 TI - [Non-invasive study of myocardial ischemia. Alternatives to the conventional stress test]. AB - Stress techniques for inducing myocardial ischemia in patients with coronary atherosclerosis have been widely studied. The most frequently used are those requiring physical effort, increasing their diagnostic efficiency with radioisotopes and echocardiographic monitoring. Patients unable to make physical exercise and with suspected ischemic heart disease may undergo nonexercise stress test. Transesophageal atrial stimulation and pharmacological stress tests with dipyridamol, dobutamine and adenosine are good alternatives, provided a strict methodology is used. PMID- 14735725 TI - [Criteria for risk of environmental pollution]. PMID- 14735726 TI - [Methodological problems in the assessment of environmental factors presenting threat to human health]. PMID- 14735727 TI - [Experience with socio-hygienic monitoring at the present stage]. PMID- 14735728 TI - [Evaluation of environmental pollution: priority trends and criteria]. PMID- 14735729 TI - [Hygienic problems of rural water supply and ways to their correction]. PMID- 14735730 TI - [Introduction of risk assessing methodology in Russia]. PMID- 14735731 TI - [Sanitary classification of enterprises and a problem of determination of sanitary-protective areas]. PMID- 14735732 TI - [Threats to environmental safety in the Russian Federation and ways of their reduction]. PMID- 14735733 TI - [Trends in water quality indices as signal of population health hazard]. PMID- 14735734 TI - [Comparative hazard of resistant organic pollutants and their substitutes]. PMID- 14735735 TI - [Substantiation of regional criteria for the safety of chemical substances for the provision of the acceptable population health hazard]. PMID- 14735736 TI - [Neglected hazard of exposure of human health to chemical substances]. PMID- 14735737 TI - [Linear dose-effect relationship for the radiation and chemical risk]. PMID- 14735738 TI - [Changes in the hydrological situation and problems of public health]. PMID- 14735739 TI - [Hygienic anthropologic pathology]. PMID- 14735740 TI - [International problems of adverse effects of asbestos and alternative fibrous materials]. PMID- 14735741 TI - [Fusaritoxins in alimentary grains and processed grain products]. PMID- 14735742 TI - [Garbage disposal in a large multidisciplinary hospital: organizational and epidemiological aspects]. PMID- 14735743 TI - [Hygienic evaluation of environmental air pollution by petroleum hydrocarbons]. PMID- 14735744 TI - [Locomotive depot as a risk factor for public health]. PMID- 14735745 TI - [Evaluation of food additives consumption in Russia]. PMID- 14735746 TI - [Non-carcinogenic risk of anthropogenic load for urban and rural area]. PMID- 14735747 TI - [The role of iodine in the formation of public health]. PMID- 14735748 TI - [Environment and public health in the area adjoining to an nuclear power plant]. PMID- 14735749 TI - [ Extreme ecology-related immunological disorders and their role in the development of nervous system diseases]. PMID- 14735751 TI - [Cancerogenic nature of ecologic toxins in cohort studies of industrial population]. PMID- 14735750 TI - [Optimization of individual health and primary prevention in the public health system]. PMID- 14735752 TI - [Problems in the diagnosis of dysbacteriosis under the influence of environmental factors]. PMID- 14735753 TI - [Biomedical criteria for the evaluation of the impact of environmental pollution on public health]. PMID- 14735754 TI - [The cytological status of the nasal and mouth mucosa for the assessment of health status]. PMID- 14735755 TI - [Drinking water nitrates and nitrites as a carcinogenic risk factor]. PMID- 14735756 TI - [Impact of emission from chemical and petroleum chemicals on public health]. PMID- 14735757 TI - [Human health and environmental factors in industrial towns]. PMID- 14735758 TI - [A new approach to diagnosing mutagenic and carcinogenic properties of environmental factors]. PMID- 14735759 TI - [A new paradigm of the role of hygienic science in the socioeconomic development of Russia]. PMID- 14735760 TI - [Functional changes in patients with arterial hypertension at high latitudes]. PMID- 14735761 TI - [Economic evaluation of population health damages caused by the influence of environmental factors]. PMID- 14735762 TI - [Appraisal of damages associated with atmospheric air pollution in the summer of 2002 to the health status of the Moscow population]. PMID- 14735763 TI - [Appraisal of the economic damage associated with cancer morbidity in Orenburg]. PMID- 14735764 TI - [Proceedings of the Plenary Meeting of the Interdepartmental Scientific Council of the Russian Federation on Human Ecology and Environmental Hygiene "Threats to Health: Topical Sanitary Problems and Ways of Their Solution". Moscow, December 15-16, 2002]. PMID- 14735765 TI - Guidelines for citing multiple authors. AB - Authors who write with others often encounter author-citation problems. Who should be and who should not be listed as an author? When is acknowledgment rather than authorship appropriate? Should the most well-known person always be listed first? How should faculty who help students publish be listed? This experienced author, coauthor, reviewer, and editor, provides answers to these questions. PMID- 14735766 TI - How can I become a journal reviewer? PMID- 14735767 TI - Mastering verb tenses in literature reviews. AB - Deciding on which verb tense to use when writing the literature review section of a manuscript is challenging. Editors find that verb tense problems are common in literature report sections of manuscripts. Authors, reviewers, and editors need to be able to spot incorrect verb tenses in literature reviews. Try editing verb tenses in the sample enclosed in this article and compare your work with that of a nursing journal editor. PMID- 14735768 TI - Hospitals' reimbursement woes continue with 2004 OPPS. PMID- 14735769 TI - JCAHO issues 'do-not-use' list of dangerous abbreviations. PMID- 14735770 TI - Recovery assistance programs help pharmacists with substance abuse problems. PMID- 14735771 TI - Bush administration rule angers lawmakers, attorneys general. PMID- 14735773 TI - Sterile compounding in hospital pharmacies. PMID- 14735772 TI - Technology helps patients with emergency room prescriptions. PMID- 14735774 TI - National survey of quality assurance activities for pharmacy-compounded sterile preparations. AB - The compliance of hospitals' quality assurance practices for pharmacy-compounded sterile preparations with the American Society of Health-System Pharmacists (ASHP) Guidelines on Quality Assurance for Pharmacy-Prepared Sterile Products was studied. A survey based on the 2000 ASHP guidelines was developed to determine current practices for compounding sterile preparations in hospital pharmacies. Practices were compared with those identified in a 1995 ASHP survey. The survey was mailed in July 2002 to a stratified random sample of 600 hospital pharmacy directors. There were a total of 182 usable responses, for a net response rate of 30.3%. Quality assurance practices for some quality domains showed low compliance with the 2000 ASHP guidelines. Only 5.2% of pharmacies that compounded risk level 1 preparations were compliant with garb requirements. Only 4.7% of pharmacies compounding risk level 3 preparations were compliant with recommendations for documentation. Compliance with guidelines on the use of resources to determine expiration dates was high (95.5%). Hospitals were moderately compliant with guidelines related to personnel training (62.7%) and labeling (61.1%). Although some improvements were observed, hospital pharmacies in 2002 were still not fully compliant with the ASHP Guidelines on Quality Assurance for Pharmacy-Prepared Sterile Products. PMID- 14735775 TI - Assessment of anticonvulsant effectiveness and safety in patients with Angelman's syndrome using an Internet questionnaire. AB - An Internet questionnaire was used to collect data from caregivers about seizure activities and drug therapies of patients with Angelman's syndrome (AS). A questionnaire to collect data on AS patient demographics, seizure types, laboratory diagnosis, and past history of anticonvulsant use was developed and distributed among the membership of multiple AS groups primarily through the Internet. Caregivers of AS patients were asked to rate each drug listed on the questionnaire for its effect on the patient's behavior and alertness and on the severity and frequency of the patient's seizures. A 6-point rating scale was used, with the lowest score representing the most favorable effect. Primary considerations of the survey were to reach as many people as possible, anonymously and quickly, in a widely comprehensible format. Data were collected between March 22 and October 9, 2002. Of the 88 complete questionnaires received, 75 were electronic and 13 were handwritten responses. Forty-seven percent of respondents reported patients' use of a combination of medications at some point, with 59 different combinations described, 21 of which consisted of three or more medications. The most commonly reported regimens were valproic acid and clonazepam, valproic acid and topiramate (with or without clonazepam), valproic acid and lamotrigine, and phenytoin and carbamazepine. Patient characteristics and seizure types were consistent with those of AS patients described in the literature. Topiramate and ethosuximide were rated favorably, consistent with previously published reports. Responses to the questionnaire were widespread and rapid, with 42 responses collected in the first week. An Internet questionnaire helped collect self-reported efficacy information from caregivers of AS patients. PMID- 14735776 TI - Recommended and actual lepirudin doses in patients with renal insufficiency. PMID- 14735777 TI - Glycemic control and medication compliance in diabetic patients in a pharmacist managed clinic in Hong Kong. PMID- 14735778 TI - Strategies to improve medication adherence in patients with depression. PMID- 14735779 TI - Changes to public health service commissioned corps. PMID- 14735780 TI - [Morphometric and ultrastructural characteristics of cardiomyocyte mitochondriome of the left ventricle in rats exposed to hypergravity]. AB - For the first time, electron-microscopic morphometric analysis of the mitochondrial system in cardiomyocytes of the left ventricle was performed in rats exposed to hypergravity (2G) After five days of exposure, the number of long mitochondria sharply increased in the interfibrillar zone of cardiomyocytes. The numbers of inter-mitochondrial junctions (IMJ) were increased in all zones of mitochondria localization. The ultrastructure and numerical density of mitochondria remained within the normal range. Similar changes were also revealed on day 19 (the end of exposure), but the numbers of IMJ in the perinuclear and subsarcolemmal perivascular zones were lower than on day 5. One months after the end of 19-day exposure at 2G, the test parameters of the mitochondrial system did not return to the norm. Apparently, this is why the repeated exposure to hypergravity (2G for five days after 30-day rest) failed to evoke a similar response from the mitochondrial system of cardiomyocytes. PMID- 14735781 TI - [Key role of apoptosis in progressing uveal melanoma]. AB - Apoptosis is known as a major achievement of biological world evolution aimed at eliminating "genetic waste," transformed or obsolete cells, and hence at controlling neoplastic processes. At the same time, clinical data point to an inverse effect. Thorough fundamental studies at the levels of light and electron microscopy allowed us to reveal several disturbances in physiological course of apoptosis. We discuss possible recruitment of this mechanism by tumor aimed at self-selection for a cell clone with maximum malignant potential and at elimination of less aggressive populations where the set of point mutations is abortive for further tumor progression. PMID- 14735782 TI - [Endogenous DNases as a tool for isolation of nuclear matrix: critical parameters of nucleolysis]. AB - Degree of nucleolysis has critical significance for isolation of nuclear matrix (NM) specifically enriched in transcribed DNA sequences as demonstrated at the example of inactive (c-fos, c-myc, and Ck) and active (p53, albumin, and 28S rRNA) genes in resting hepatocytes. Optimal degree of nucleolysis features degradation of loop domains of chromatin with preserved relatively uniform molecular weight distribution of DNA. Deviation from these parameters leads to nonspecific fragmentation of chromatin in various gene loci and isolation of NM samples nonspecifically enriched or depleted of transcribed DNA sequences. Under optimal hydrolytic conditions, the transcribed chromatin is more resistant to endogenous DNase attack, which allows selective conservation of its association with the nuclear matrix. PMID- 14735783 TI - [Calcium and ammonia stimulate monoamine oxidase A activity in brain mitochondria]. AB - The effect of ammonia and calcium on the activity of monoamine oxidase (MAO) was studied. The enzyme activity in non-synaptic brain mitochondria isolated from rats administered with ammonium acetate was assayed by release of H2O2 using spectrophotometry. The effect of calcium on MAO was assayed directly after addition of Ca2+ to the non-synaptic mitochondria isolated from the forebrain of the control rats. Both ammonium acetate injection in vivo and Ca2+ addition in vitro stimulated the activity of MAO A but not that of MAO B in mitochondria. This is the first evidence that ammonia and Ca2+ regulate MAO A in non-synaptic mitochondria in the forebrain and can contribute to oxidative stress in the neurons via MAO A activation. PMID- 14735784 TI - [Chemical composition and cryoprotective activity of ethanol extract from winter caterpillars Aporia crataegi L]. AB - Glycerol, alpha-carotene, and other yet unidentified compounds have been found in the ethanol extract from winter caterpillars of the black-veined white (Aporia crataegi L.). We have shown that the ethanol extract has a cryoprotective effect on human peripheral blood lymphocytes, and this activity is approximately three times higher than that of glycerol (particularly, in the case of repeated freezing), which is one of the best cryoprotectors. PMID- 14735785 TI - [Study of degradation of flavonols in the mutants of poppy (Papaver somniferum L.)]. AB - We studied flavonol-degrading activity of cell-free extracts from petals of the flower color and structure mutants. The relationship between degradation of flavonols (kaempferol, quercetin, and myricetin) and biosynthesis of anthocyanins has been revealed. The highest flavonol-degrading activity has been revealed in white flower mutants towards all substrates, particularly, quercetin. The mutations inhibiting synthesis of an anthocyanin pelargonidin provide for synthesis of various quantities of cyanidin in the petals. The flavonol-degrading activity considerably increases with the content of cyanidin. A similar relationship has been revealed in the mutants synthesizing both cyanidin and pelargonidin. The plants accumulating considerable quantities of pelargonidin in their petals have accordingly higher flavonol-degrading activity and predominantly hydrolyze kaempferol. The plants forming additional pods in their flower (pistillody) have higher flavonol-degrading activity as compared to the anther-in-petal and doubleness mutants. PMID- 14735786 TI - [Effect of oligosaccharides on adaptation of winter wheat seedlings to negative temperature]. AB - We studied the dynamics of endogenous content of bioactive oligosaccharides in the roots of winter wheat seedlings. Previously these oligosaccharides proved to mediate development of frost resistance during the first days of hardening (Zabotina et al., 1998). The changes in their endogenous content can be described by a single-humped curve peaking 6 h after the onset of frost hardening. The capacity of these polysaccharides to increase frost resistance (LT50 was evaluated by leakage of electrolytes) when added to growth medium did not depend on the pretreatment duration (from 1.5 to 18 h) but decreased if they were introduced in the course of the adaptive response. Inhibition of the adaptive response by inhibitors of RNA and protein synthesis was ceased in the presence of the oligosaccharides. We believe that the oligosaccharides that are products of metabolism of the cell wall polysaccharides are involved in adaptation to low temperature. PMID- 14735787 TI - [Pattern of selenium distribution in tomatoes Lycopersicum esculentum Mill]. AB - We studied the pattern of selenium accumulation in Lycopersicum esculentum Mill plants in conditions of root and extra-root dressing with selenium. At the background of general trend towards predominant selenium accumulation in the fruit coat, a higher level of the microelement accumulation in the pulp was observed for selenates (Se+6) than selenites (Se+4). Application of Epin growth factor favored selenium accumulation in the plants; its concentration in the pulp was comparable to the concentration after selenate application, while the level of the microelement accumulation in the fruit coat, leaves, and roots was 2.2, 2.4, and 1.4 times lower, respectively. Combined application of Epin and sodium selenate had virtually no effect on selenium accumulation in the pulp but increased the microelement content in the fruit coat and roots. Biological significance of the observed "boundary effect" of selenium as a manifestation of plant antioxidant protection is discussed. PMID- 14735788 TI - [Transport of ions in mitochondria in connection with osmotic concentration of media]. AB - Studies on mitochondria has revealed apparent violation of laws of osmosis, apparently, related to ion transport. In this context, we studied changes in the content of potassium, sodium, magnesium, and calcium in mitochondria from wheat (Triticum aestivum L.) roots incubated in the buffer without the substrate with 0.3 or 0.5 M sucrose as well as potassium, and phosphate. The most pronounced changes were observed for potassium. Potassium arrival to mitochondria was more active at 0.5 than 0.3 M sucrose. The direction of the cation transport depended on its content in the initial buffer. At the certain threshold level, the inflow was replaced with the outflow; this threshold was lower at 0.3 than 0.5 M sucrose. Magnesium was also released from mitochondria; its stable outflow was primarily observed at its lower initial content in the suspension at 0.3 rather than 0.5 M sucrose. The obtained data demonstrated that the apparent violation of laws of osmosis was false and resulted from ion transport. Passive ion transport as a possible mechanism of adaptation to osmotic properties of the external medium is discussed. PMID- 14735789 TI - [Food energy expenditure by the moth Lymantria dispar L. (Lepidoptera, Lymantriidae) at different stages of ontogenesis]. AB - Food energy expenditures by gypsy moth at different developmental stages under conditions of feeding on larch needles are described. The distribution of total food energy (%) for metabolism, the formation of exuvia (throughout ontogeny), and the formation of an adult insect is assessed. The proportion of food energy lost with excrements over the entire period of feeding is the same (58%) in females and males. The assimilated part of food energy is distributed differently: energy expenditures for the formation of exuvia and adult insects are greater in females than in males. The proportion of food energy expended for metabolism throughout the insect life span is approximately 30% in both females and males. PMID- 14735790 TI - [Effect of monochromatic light on the growth and development of larva of the brown frog Rana temporaria L]. AB - Acceleration of the growth and development of R. temporaria larvae under the effect of short-wavelength light was observed in experiments. However, the highest linear and weight parameters of postmetamorphic juveniles were recorded upon exposure to yellow and green light, and the lowest parameters, upon exposure to blue light. The red zone of the light spectrum had no apparent effect on the development of tadpoles and juvenile frogs. PMID- 14735791 TI - [Psychophysiological study of the information theory of emotions using the model of positive emotion]. AB - The pattern of inter-hemisphere distribution of EEG amplitude and frequency as a function of the levels of emotional experience and motivation as well as probability of the goal achievement was studied in 20 subjects. An emotional state was evoked by simulating emotionally colored events. A modified test of Prise et al. (1985) was used to evaluate the level of motivation for the simulated event as well as the probability of goal achievement from the lengths of line segments marked by the subjects. Here we studied simulated emotion of joy. The highest correlation coefficients were observed between the awareness and alpha activity in the both hemispheres. The levels of emotional experience and motivation inversely correlated with the delta and theta activity mostly in the left hemisphere. The beta activity correlated with both the emotional and motivation levels. PMID- 14735792 TI - [Role of glycine in blood coagulation processes]. AB - We studied hemocoagulation properties of the amino acid glycine in vitro and after intravenous administration in animals (rats). Addition of 10(-3) and 10(-4) M glycine to the plasma increases the aggregability of thrombocytes in vitro, while all other tested concentrations had virtually no effect on hemostatic parameters of the plasma. Intravenous administration of glycine increased functional activity of the enzyme fibrinolytic unit of the anticoagulation system. Dose dependence of this effect has been established. The causes of these changes and possible application of this compound for activation of fibrinolysis are discussed. PMID- 14735793 TI - [Indication of Northern Caspian regions by biochemical (enzymatic) indices of water and bottom sediments]. AB - We evaluated ecological status in the region of mixing of river and sea waters in the northern Caspian Sea with high anthropogenic pressure. We studied the regions of Volga River fore-delta, the water areas neighboring ancient hydrological transects, and the sea regions between the transects. The water areas were evaluated by biochemical degradation of organic compounds (proteinase and amylase activities) in water and bottom sediments as well as hydrochemical indices of water (salt composition and contents of dissolved oxygen and biogenic compounds). In conditions of active vegetation of phytoplankton and higher aquatic plants soon after the highest wave of the Volga flood, we studied distribution of hydrolytic enzyme activities involved in the global production/destruction cycle of high molecular weight organic compounds (proteins and polysaccharides). The data obtained with the trypsin-azocasein and alpha-amylase-modified starch tests indicates pollution of water and surface sediments in the lower reaches of Volga and the inflow sea regions. PMID- 14735795 TI - [Effect of high intensity magnetic field on the processes of early growth in plant seeds and development of honeybees]. AB - The influence of magnetic field on the early growth processes in plant seeds and the postembryonic development of honeybees was studied. Some general trends in the effects of magnetic field and differences in the tolerance of plant seeds and developing honeybees to its action were revealed. Some factors that may be responsible for a low reproducibility of magneto-biological effects are discussed. PMID- 14735794 TI - [Early toxic effect of zinc, cobalt, and cadmium on photosynthetic activity of green alga Chlorella pyrenoidosa Chick S-39]. AB - Early toxic effects of heavy metals (HMs) Zn, Co, and Cd in concentration from 0.01 to 100 mM on photosynthetic activity of the green alga Chlorella pyrenoidosa Chick S-39 were studied. The early effect of HMs was manifested as a rapid (0.5-2 h) reduction of photoinduced oxygen release by the algal cells. The suppressed relative yield of variable chlorophyll fluorescence ratio (Fv/Fm) by the metals as well as its dynamics in C. pyrenoidosa demonstrated rapid inactivation of photosystem II (PS II). Analysis of the induction curve of delayed chlorophyll fluorescence in Chlorella cells suggested that the early toxic effects of the tested concentrations of Zn, Co, and Cd included both reduced electron transport in PS II and decreased photosynthetic membrane energization. Hence, the early toxic effect of Zn, Co, and Cd was primarily related to the decreased efficiency of the light reactions of photosynthesis which further reduced the alga productivity later. PMID- 14735796 TI - [Ecological and biological features of soils in the forests defoliated by the siberian moth in the southern taiga of middle Siberia]. AB - Experimental data are analyzed that concern the effect of zoogenic debris on the properties of soddy deep podzolic soils and raw-humus brown soils characteristic of southern taiga forests in the Yenisei region of Siberia. It is shown that the influence of excrements of Siberian moth larvae on the soil microflora lasts for two or, at most, three growing seasons. Zoogenic plant debris falling on the ground surface during tree stand defoliation is a short-acting but powerful stimulant of biological activity in the litter; hence, it has a considerable effect on soil properties. This effect is enhanced by changes in ecological conditions that occur upon defoliation. The influence of the cenotic factor on biogenic soil properties is manifested more strongly in the organogenic horizons. The communities of microorganisms involved in the nitrogen and carbon cycles are dominated by prototrophic forms in the normal fir forest and by pedotrophic forms in the forest defoliated by pests. PMID- 14735797 TI - [History of marine microbiology in Russia (USSR) in the second half of 20th century]. PMID- 14735798 TI - Revalidation of Sri Lankan doctors. PMID- 14735799 TI - Suicide prevention strategies in Sri Lanka: the role of socio-cultural factors and health services. PMID- 14735800 TI - Cost accounting in a surgical unit in a teaching hospital--a pilot study. AB - INTRODUCTION: Economic constraints remain one of the major limitations on the quality of health care even in industrialised countries. Improvement of quality will require optimising facilities within available resources. Our objective was to determine costs of surgery and to identify areas where cost reduction is possible. PATIENTS AND METHODS: 80 patients undergoing routine major and intermediate surgery during a period of 6 months were selected at random. All consumables used and procedures carried out were documented. A unit cost was assigned to each of these. Costing was based on 3 main categories: preoperative (investigations, blood product related costs), operative (anaesthetic charges, consumables and theatre charges) and post-operative (investigations, consumables, hospital stay). Theatre charges included two components: fixed (consumables) and variable (dependent on time per operation). RESULTS: The indirect costs (e.g. administration costs, 'hotel' costs), accounted for 30%, of the total and were lower than similar costs in industrialised nations. The largest contributory factors (median, range) towards total cost were, basic hospital charges (30%; 15 to 63%); theatre charges fixed (23%; 6 to 35%) and variable (14%; 8 to 27%); and anaesthetic charges (15%; 1 to 36%). CONCLUSION: Cost reduction in patients undergoing surgery should focus on decreasing hospital stay, operating theatre time and anaesthetic expenditure. Although definite measures can be suggested from the study, further studies on these variables are necessary to optimise cost effectiveness of surgical units. PMID- 14735802 TI - Preconceptional care of women at booking visit at De Soysa Maternity Hospital and Castle Street Hospital for Women. AB - OBJECTIVE: To study the preconceptional preparedness of women attending two antenatal clinics. DESIGN: Cross-sectional descriptive study done in August and September 2001. SUBJECTS AND SETTING: Pregnant women attending the antenatal clinics De Soysa Maternity Hospital and Castle Street Hospital for Women for their booking visit. MATERIAL AND METHODS: Pregnant women were randomly selected. Before collecting data the purpose of the study was explained and those who consented were recruited for the study. Data were collected on the basis of an interviewer administered questionnaire. Ethical approval was obtained from the Ethical Review Committee of the Faculty of Medicine, University of Colombo. RESULTS: 225 pregnant women were recruited. 55% of them were between the ages of 18 and 30 years. 96% had achieved an educational level of above year five. 55% were in their first pregnancy and 2.75 were grand-multipara. 186 (82.7%) were housewives. 81% had a planned pregnancy. Only 21% had received pre-pregnancy counselling, 52% of them from a specialist obstetrician, and 21% and 19% from a general practitioner and public health midwife. Only 15 (6.6%) had taken preconceptional folic acid supplementation, and all of them had a level of education of GCE (A/L) or above. 11 of those who took preconceptional folic acid were primipara. 159 (70.6%) had received rubella vaccination. Of those who did not take the vaccine, 44% knew about it but did not know its importance, and 38% did not know about its availability. 18% did not take it because of various myths that they believed in. Preconceptional health knowledge regarding pregnancy was assessed by asking 10 questions and expressing it as a score out of 10. This score showed a positive correlation to the level of education of the woman. A majority received information from the print (81.7%) and electronic (72.4%) media. 50% received information from a public health midwife, and 36% from doctors. CONCLUSIONS: Preconceptional preparedness among our women is poor. However, rubella vaccination is relatively successful compared to other aspects of preconceptional preparedness. PMID- 14735803 TI - Caring for the dying patient. PMID- 14735801 TI - Knowledge on lymphatic filariasis and the response to July 2002 mass treatment campaign in two communities in the Galle district. AB - OBJECTIVE: To ascertain the knowledge on filariasis and response to the July 2002 mass treatment campaign in two sample populations. DESIGN: Application of pre tested questionnaire by direct interviews of individuals from randomly selected streets. METHODS: Study areas were a coastal community in Unawatuna (population sample 381), and an inland community in Baddegama (population sample 236) in the Galle district. They were interviewed twice, 4 weeks before the mass drug administration (MDA) and 4 to 7 days after. RESULTS: The sample population of Unawatuna had a greater awareness of the clinical and parasitological features of the disease (p = 0.0003) and the drug treatment (p = 0.00380 than that of Baddegama. Only 5.5% of the combined sample attributed the cause of filariasis to a parasitic worm. However, over 70.0% of them knew that transmission was through mosquito bites. Volunteers formed 87.5% of the work force used for drug distribution in Unawatuna and 70.1% in Baddegama. The balance work force were Grama Niladaris and Public Health Midwives. Drugs were received by 76.9% of the Unawatuna sample compared with 89.0% at Baddegama (p < 0.001). Among those who received the drugs, consumption was 91.8% in Unawatuna and 96.2% in Baddegama. Taking the two communities together adverse effects were experienced by 22.9%. These effects in night time drug consumers (10.4%) were significantly less than in day time consumers (19.8%) (p < 0.005). The adverse effects were sleepiness (37.8%), malaise (28.2%), headache (16.8%), vomiting (5.1%), nausea (4.5%) and fever (3.9%). The message of mass treatment was carried to the community by the people themselves on the eve and on the day of drug distribution. Over 95% agreed that in future programs drugs should be delivered to their homes. PMID- 14735804 TI - The Sri Lankan symphysiofundal height chart is misleading. PMID- 14735805 TI - Adverse drug reactions in children requiring hospital admission. AB - OBJECTIVE: To study the incidence of adverse drugs reactions (ADRs) in children admitted to Lady Ridgeway Hospital (LRH), Colombo. DESIGN: A prospective hospital based descriptive study. SETTING: Medical units of LRH, Colombo. METHODS: Information was collected by a hospital based investigator who visited the medical units at LRH from February to December 2002. The Naranjo algorithm was used to classify the information. REMARKS: During the 11 month study period, 63 admissions were due to ADRs, the commonest being secondary to administration of antibiotics. CONCLUSIONS: ADRs accounted for only 0.16% of admissions as compared to about 2% in North America. The commonest ADRs were due to antibiotics. PMID- 14735806 TI - Direct oxidant damage to red cells associated with propanil ingestion. PMID- 14735807 TI - Ohtahara syndrome. PMID- 14735808 TI - Pseudo-convulsions in a child subjected to abuse. PMID- 14735809 TI - Percutaneous transluminal septal myocardial ablation in hypertrophic obstructive cardiomyopathy. PMID- 14735810 TI - A case of hypophosphatasia. PMID- 14735811 TI - Preponderance of blood group B among dengue fever patients with serious complications in a tertiary care hospital. PMID- 14735812 TI - Delusional parasitosis. PMID- 14735813 TI - Hookworm infection as a cause of melaena. PMID- 14735814 TI - A pilot study of a low cost CLO test. PMID- 14735815 TI - Salmeterol multicentre asthma research trial (SMART): interim analysis shows increased risk of asthma related deaths. PMID- 14735816 TI - Reducing abortions is a public health issue. PMID- 14735817 TI - The changes in major limb reconstruction as a result of the development of allografts. AB - Over the past 30 years the author and his colleagues have performed almost 1100 non-pelvic massive cadaveric allografts mostly in the treatment of malignant tumors of bone. The overall results show that 77% of the devices are still functional and competent. The best results were the intercalary grafts (86%) while the poorest were the allograft arthrodeses (61% successful). The complications of fracture and non-union are the principal problems reducing the success rate to 48% and 71% respectively. Primary infection was a major issue but occurred in less than 8% of the cases and reduced the success rate to 72%. Secondary infection following operative procedures to repair fracture or graft non-union occurred in 53 cases and reduced the success rate to less then 30%. No patients died of infection. PMID- 14735818 TI - The results of the surgical treatment of bone tumors using massive homoplastic grafts. AB - We reviewed our experience as concerns 588 patients treated by resection for bone tumor and reconstructed by massive bank bone implant. The period taken into consideration goes from 1984 to 2001. The mean age of the patients was 24 years with a minimum of 1 and a maximum of 77 years: in 186 cases (31.6%) age was lower by 14 years. Most of the patients were referred for a diagnosis of malignancy (85%), and of these, 70% had undergone antiblastic therapy postoperatively according to the modalities of the protocols. Of the cases that were not treated by chemotherapy there are 16 that had a homoplastic graft after failure of a previous one. In a high number of cases (189) surgery included reconstruction of an intercalar segment of a long bone. Of these, 106 were in the femur, 69 in the tibia, and 14 in the humerus. One joint reconstruction was carried out in 128 cases of which 52 in the knee, 49 in the shoulder, 10 in the elbow, 11 in the wrist, 3 in the ankle, and 3 in the metatarsals. Fusion, which was one of the most frequent indications in the eighties, is used less today; overall it was carried out in 91 cases of which 65 in the knee, 14 in the hip, 9 in the ankle and 3 in the wrist. This type of implant may also be used in the pelves, and this took place in 56 cases using different procedures of reconstruction. The most current method of reconstruction for a joint is composite prosthesis, and this was used in 124 cases of which 67 around a knee, 53 in the hip, 4 in the shoulder. Finally, in 78 cases a vascularized fibula transplant was associated with an intercalar graft, fusion, or osteoarticular graft to increase the mechanical capacity of the implant together with a more rapid integration of the implant, particularly at the osteotomic lines. Despite the excellent results demonstrated in the first 3 years of follow-up, infection, resorption, delay in consolidation and fracture led to 18% failures of the implant. Massive homoplastic graft is still a good means of reconstruction that allows for excellent functional results, but despite this intense research activity is required to improve its mechanical hold in time together with better and complete integration. PMID- 14735819 TI - Knee resection arthrodesis with allograft: a long-term follow-up study. AB - A consecutive series of 57 patients treated by knee resection arthrodesis for malignant or aggressive tumor around the knee was reviewed. Infection was present only after repeated surgery for other complications, delayed union or non-union occurred in 50% of the cases that could be evaluated, but were still easy to manage. Fracture incidence was higher than expected (32.6%) even occurring after 10 years; this was difficult to deal with and it often led to failure. The best possible method of fixation is still being debated, but locked nail and allograft cementation is often advised. Several satisfactory functional results were however achieved when surgery was performed in young patients; final results can be less satisfactory when there is leg length discrepancy and poor acceptance on the part of the patient. In recent years this type of surgery has been limited to younger male patients (10 to 14 years of age) in whom extra-articular knee resection was required or when most of the quadriceps muscle must be removed. PMID- 14735820 TI - The role of vascularized fibula in skeletal reconstruction. AB - Since March 1988, 144 oncologic bone resections have been surgically treated with vascularized fibula (VF). In 30 patients VF was used after the failure of previous reconstructions, while in 114 cases (79%) VF immediately followed tumor resection. In 14 cases VF was pedicled on its vessel and used to reconstruct the ipsilateral tibia. In 17 young children, the proximal fibula was used to reconstruct with a potential growth the proximal humerus, the distal radius or the proximal femur. In 103 cases VF was combined with bone allografts. In 70 of these the fibula was inserted inside the massive allograft as a viable rod. There were 7 mechanical failures, 9 deep infections, and 10 local recurrences. According to functional evaluation, 69 patients (48%) presented with excellent functional results, 39 patients (27%) had good results while fair of poor results were recorded in 36 cases (25%). PMID- 14735821 TI - Survival of 25 osteoarticular allografts followed more than 10 years. AB - An evaluation of the long-term survival rate of 25 osteoarticular allografts was made. Clinical analysis was based on the Mankin scale and the Musculoskeletal Tumor Society (MSTS) Grading system. After a mean follow-up time of 15 years 76% of the osteoarticular allografts had good or excellent rating. The MSTS scale revealed a mean score of 89% for those 20 grafts still functioning according to their primary purpose. Allograft related complications occurred in 32% of the cases being most common among malignant cases. Due to the rather good long-term results, osteoarticular allografts can still be recommended for hemicondylar allograft reconstruction in benign lesions. PMID- 14735822 TI - Pulse lavage washing in decontamination of allografts improves safety. AB - We analyzed the bacterial contamination rate of 140 femoral head allografts after rinsing the allografts in different decontamination solutions. Bacterial screening methods and cleansing effect of antibiotics (cefuroxime and rifampicin) and pulse lavage were compared. Swabbing and taking small pieces of bone for culture were the screening methods used. Both methods proved to be quite unreliable. Approximately one-fourth of the results were false negative. Culturing small pieces of bone gave the most accurate and reliable results and, therefore, can be recommended as a bacterial screening method. The use of antibiotics in allograft decontamination is controversial. In prophylactic use antibiotics include risks of allergic reactions and resistant development and our results in the present study show that antibiotics do not improve the decontamination any better than low-pressure pulse lavage with sterile saline solution. Therefore, pulse lavage with sterile saline solution can be recommended for allograft decontamination. Our results demonstrate that it decreases bacterial bioburden as effectively as the antibiotics without persisting the disadvantages. PMID- 14735823 TI - Total hip replacement after tumor resection--30 years experience. AB - The authors report their experience with 161 cemented total hip replacements after tumor resections around the hip implanted between 1971 and 2001 in patients from aged 10 to 79 years. At first, the long stem Protek module was used and since 1980 the Czech made long stem Poldi endoprosthesis has been implanted. A cemented UHMW polyethylene cup has been used with both stems. 66 THR were implanted for primary tumors from which 47 were malignant. 3 Protek and 2 Poldi stems fractured. Early dislocation was common with standard cups; therefore the authors recommend anti-luxation cups with higher rims in all surgeries to enable early ambulation. They also observed better long-term results with the cup slightly in varus. Multiple reimplantations are to be expected when THR is implanted before skeletal maturation, but they are not a problem. The Poldi model is an excellent cost effective solution, especially for secondary malignant tumors around the hip. In the sixties hemiarthroplasty was usually the only chance for a good functional salvage after tumor resections of the proximal femur. Since the seventies special long stems have been developed. Continuous follow up of these surviving patients is important for further development of this special field of endoprosthetics. PMID- 14735824 TI - Bone metastases in the hip region: surgical treatment. AB - Subjective factors of the patient such as age, weight, occupation, expectations, etc. must be carefully evaluated in terms of surgery. It is always important to remember life expectancy, and the choice of a surgical procedure must be based on it. Limited survival and possible radiation therapy must eliminate methods that require long healing times: we no longer use bone grafts because the risk of healing time taking up too much of the time left for the patient is too high. Reconstructions must have an immediate solidity allowing early use of the limb. Thus, in the proximal femur, we above all use protheses (traditional or modular RPS); in acetabular lesions that do not involve the joint surface, our treatment of choice is curettage with liquid nitrogen and cement, armed with screws and nails. Prostheses, which are always cemented, are assembled using bipolar cups; cases where lysis is so wide on the cotyloid side as to require THR are rare. The cotyles, in cases such as these, are those that allow for anchoring with several sites (Octopus type), the McMinn or, at times, common cotyles in PE. PMID- 14735825 TI - Resection of the proximal humerus for metastases and replacement with RPS prosthesis. AB - In the surgical treatment of metastatic lesions of the proximal humerus it is important to find solutions that eliminate pain and solve the mechanical problem in a short time and with reduced costs. In 90% of cases, intralesional curettage, freezing with liquid nitrogen and cement enhanced intramedullary instrumentation is capable of brilliantly solving the problems of these patients. When, instead, osteolysis involves the greater tuberosity and/or the joint region, resection of the proximal humerus followed by reconstruction with a modular prosthesis is indicated. Of those available on the market, the RPS system (LIMA) has features that make it equivalent to others, but at costs considerably lower. However, in reconstructions with prostheses, active abduction is significantly limited. A personal series of 20 patients is presented. PMID- 14735826 TI - Use of the Ilizarov technique to improve limb function following hemipelvectomy. A case report. AB - Various surgical methods have been used for reconstruction after limb salvage surgery for pelvic tumours. We present a case of use of the Ilizarov technique as a second stage adjunct, for the improvement of function, after first stage tumour resection and arthrodesis. This technique is suitable for selected patients with tumour-free survival following the initial resection. Though not described before, our experience indicates it is an alternative to a mammoth one stage internal hemipelvectomy with reconstruction, that is often attempted even though there is a high risk of local recurrence and distant metastasis. PMID- 14735827 TI - Current trends in chest wall malignant tumor. AB - Surgery used to treat chest wall sarcomas requires preoperative evaluation in order to document their margins, and to reveal any nodules of recurrence. Because of its considerable sensitivity and specificity ultrasound was associated with routine imaging techniques. The results obtained are useful in defining superficial and lateral tumor margins, but particularly in determining micro nodules of recurrence which would otherwise go unrecognized. Based on the data obtained using routine methods of diagnosis and ultrasound, we submitted patients to excision that each and every time proved to be wide on histological examination. A particular surgical method was used for reconstruction, involving a polypropylene mesh fixed to one or two mouldable plates. This method exploits the elasticity of polypropylene, which is indispensable for the correct respiratory dynamics of the rib cage, thus avoiding paradoxical or harmful movement. Oncological and functional results were satisfactory. PMID- 14735828 TI - Substitution in vertebral resections. AB - The authors discuss the reconstructive methods used after curettage and/or vertebral resection possibly associated with removal of surrounding muscular, visceral and nervous structures. PMID- 14735829 TI - A model for the organization of a regional bank for musculoskeletal tissues in Italy. AB - The national and local need for human tissues to be used in transplants is a high one, and constantly growing. Human, scientific and financial resources involved in guaranteeing safe and high-quality tissues as defined by the national guidelines for musculoskeletal tissue banks are considerable. For this reason we need to find adequate solutions to the problem of guaranteeing sufficient availability of tissues with the lowest cost possible for supply. The Piedmont Region Musculoskeletal Tissue Bank, which is located in an Azienda Ospedaliera, has been organized to guarantee the quality of its tissues via biological validation and scientific-clinical coordination; it makes use of the collaboration of removal centers that send tissues taken from the central bank for certification and identifies several centers for preservation; it collaborates with a bank of national importance for tissue processing. The publication of regulations and tariffs based on cost analysis improves the procedures. PMID- 14735830 TI - The role of basic research in clinical treatment of Ewing's sarcoma and osteosarcoma. PMID- 14735831 TI - A computer assisted approach for monitoring and planning biological reconstructions in orthopaedic oncology: preliminary results and future perspectives. AB - The evaluation of the mechanical strength of a reconstructed bone, and its evolution during the follow-up is, at present, almost impossible. This information, however, may prove extremely useful in clinical practice, both in the surgical planning and in the management of the rehabilitation therapy. In this work, a non-invasive technique, based on the use of finite element modelling, is presented that allows the simulation of the mechanical behaviour of a skeletal reconstructions starting from Computed Tomography data. This method was applied to study the evolution, during the first year of follow-up, of the strength of the reconstructed femur of a ten year-old child if a short, slow, level walk were allowed. The preliminary results indicate that the window sculpted by the surgeon to allow the anastomosis of the vascular pedicle acts as a stress concentrator and that the reduced bone mineral density, induced in the child's bone by the absence of load during the post-operative period, increases the stress level in the proximal femur. PMID- 14735832 TI - The integration of vascular surgical techniques with oncological surgical protocols in the treatment of soft tissue sarcomas of the limbs. AB - Soft tissue tumors, involving the vascular bundle, require a particular surgical approach: oncological and vascular surgical techniques must be integrated in order to perform a limb-saving surgery with adequate margins. Thirty-six soft tissue sarcomas of the thigh and popliteal region were treated from June 1999 to September 2002. Nineteen cases involving the vascular bundle were analysed and placed in two groups according to imaging and clinical information: Group A, 14 patients, with tumors close to femoral vessels without adventitial infiltration, and Group B, 5 patients, with vascular infiltration. Group A was treated with vascular blunt dissection performing adventitial excision. Group B was treated with vascular "en-bloc" resection and reconstruction. Imaging and clinical information together with surgical techniques, strategies and complications were analysed in order to plan the surgical approach in neoplastic vascular bundle involvement. PMID- 14735833 TI - Echocolor Power Doppler with contrast medium to evaluate vascularization in lesions of the soft tissues of the limbs. AB - PURPOSE: Echocolor Power Doppler with contrast medium forms a non-invasive vascular image; the purpose of the study is to evaluate the effectiveness in differentiating benign and malignant tumors in the soft tissues of the limbs. MATERIAL AND METHOD: Echocolor Power Doppler with contrast medium was used to study 80 patients with swelling in the soft tissues of the limbs: there were 54 benign lesions, 22 sarcomas, and 4 aggressive desmoid fibromatoses. RESULTS: Were identified 4 patterns of wash-in and wash-out curves that could be correlated to the histological diagnosis: type I was present in 85% of benign lesions, type III in 91% of malignant lesions and in 3.7% of the benign ones, type II in aggressive fibromatoses, anomalous type in 4 benign lesions and 2 sarcomas; the curve was absent in 2 benign lesions. CONCLUSIONS: Power Doppler Echocolor with contrast medium can become a useful method to be associated with traditional imaging methods in the differential diagnosis of swelling of the soft tissues of the limbs. PMID- 14735834 TI - CT evaluation pre- and post-percutaneous ablation by radiofrequency of osteoid osteoma. Preliminary experience. AB - PURPOSE: Evaluation of bone remanagement after treatment by thermoablation of osteoid osteoma (OO) by CT scan. MATERIAL AND METHOD: Nine cases of OO (8 in the limbs, 1 in the pelvis) following biopsy were treated by CT-guided thermoablation. Clinical results, complications, density of tissues treated by CT scan (pre-postop, 6, 12 months) are evaluated. RESULTS: Absence of complications, regression of pain over 2 weeks, resumption of sports activity in 1 month. Bone density after treatment increases but even after 1 year it is much lower than normal levels. CONCLUSIONS: Bone remodeling after thermoablation of OO requires much time, the process is still visible 12 months later by CT scan. CT scan is an adequate method, not only for diagnosis and treatment, but also for follow-up, capable of evaluating in time the changes in density of the site of the lesion, which is useful for a comparison in case of postoperative pain of doubtful origin. PMID- 14735835 TI - [Death caused by the heat wave: epidemic that should not be forgotten]. PMID- 14735836 TI - [Heat and health: effectiveness and equity]. PMID- 14735837 TI - [Assessment and prevention of the health effects of the climate: an European project and a national project]. PMID- 14735838 TI - [Health profile of a population as a predictor of health resources utilization: results of the implementation of an epidemiologic model for needs analysis]. AB - The integration of the scientific researches in the assistance process is one of the most important challenges that is currently set to the health operators. In this paper a model for the health needs-assessment will be applied to verify if and how the prevalence of some classical risk factors for cardiovascular disease foretells mortality and hospitalisation episodes at 3 years, and if it could express the health need of that population. The "sanitary history" of 1704 subjects, enrolled in 1996 during the Brisighella Study, has been followed. We know the health profile of these subjects at 1996 and data about their hospitalizations, mortality, and general assistance from 1996 to 1999. In this population the risk of cardiovascular disease is inferior to that esteemed by the hospitalisation rate, attributable mostly to a little group of subjects with well defined characteristics of exposure. The resources spent on a hospitalization do not adequately describe either the sanitary need nor the relief load and the "cost" associated to the disease. The methodology used allows to explore in detail the relative weight of the different subjects involved in the sanitary assistance in order to better reach the objective of producing the maximum quantity of benefits for the patient at the smallest possible quantity of risk. PMID- 14735839 TI - [Assessment of risk of mesothelioma: the case of an asbestos-cement production plant in the city of Bari]. AB - OBJECTIVE: In this paper, by means of explorative methods based on geographical analysis, the relationship between the presence of an asbestos-cement factory in the urban area of Bari and malignant mesothelioma cases, occurring between 1980 and 2001 among residents, is analysed. METHODS: The data source of the 64 cases studied is the national register of mesotheliomas (Renam), the Apulia regional operating centre (Cor-Puglia). Data are analysed by the use of the S + SpatialStats software. RESULTS AND DISCUSSION: Both individual data analysis and explorative geographic analysis point out an increased risk of disease among people living near the asbestos-cement factory: within an area centred on the location of plants and having a radius of about 1 Km, the estimated risk is 2.38 times above the normal level. Further analytical studies are required. PMID- 14735840 TI - [Impact of smoking in Italy in 1998: deaths and years of potential life lost]. AB - This report presents the impact of smoking habits on Italian mortality in 1998. Estimates of smoking-attributable fraction (FAF), smoking-attributable mortality (MAF), and years of potential life lost (YPLLf) were calculated using the SAMMEC software (CDC, USA), and the Peto method. During 1998, using the SAMMEC software, smoking caused approximately 83,650 premature deaths in Italy (67,600 in men, 16,000 in women, and 45 in infants), equal to 15.1% of mortality in adults above the age of 35 years (24.4% of mortality in men and 5.8% in women). YPLLf were about 900,000 in men, 221,000 in women, and 3,500 in infants. Using the Peto method, smoking-attributable deaths were about 70,200 (59,600 in men and 10,600 in women), equal to 12.7% of mortality in adults older than 35 years (21.5% of mortality in men and 3.9% in women). YPLLf were about 806,000 in men and 142,000 in women. Among adults, for both methods most smoking-related deaths were attributable to lung cancer, ischemic heart disease, chronic airways obstruction, and cerebrovascular disease. Differences between the estimates of the two methods underline the gap between a more conservative estimate (Peto method) and a less restrictive one (SAMMEC software). Given validity for both methods, it is likely smoking habits caused between 70,000 and 83,000 deaths in Italy during 1998. PMID- 14735841 TI - [Increase of births by cesarean section in Campania in 2000]. AB - Cesarean deliveries (CS) are increasing in Italy, especially in the Campania region, where they reached a peak incidence of 52% in the year 2000. This study analyses CS rate variability by hospital type, foetal risk factors (low birth weight, prematurity, plurality, anomalous presentations), maternal characteristics (age and parity) and delivery time. Data are drawn from birth certificates of 31,182 babies, that is 64.2% of all delivered in 65 out of 98 Campania birth-places. 16,251 (52%) babies are born by CS; 559 (1.8%) by operative vaginal delivery. The Attributable Risk for foetal risk factors is 6.9%. In absence of these the CS rate would be 48.5% instead of 52%. CS rate of primigravid women was 53.2%. 63% of CS take place during morning hours and 93% on working days. CS rate is higher in private than in public hospitals (58 vs 46%), though foetal risk factors are lower. Rate of older women was higher in public hospitals, rate of primigravid women was higher in private ones. Campania data confirm the marginal role of foetal risk factors in the increase of CS rate. In order to reduce CS rates it seems reasonable to offer special support to primigravid low risk mothers and to provide behavioural obstetrical guidelines for the peculiar Italian reality. PMID- 14735842 TI - [Twins in biomedical research and the creation of the "National Twin Registry"]. AB - Twins are a valuable resource for the study of complex traits. The twin method is substantially based on the comparison between correlations and concordance in monozygotic (MZ) and dizygotic (DZ) twins and allows several applications in biomedical and molecular genetic research. It allows either the qualitative and quantitative evaluation of the influences that genetic and environmental factors exert on phenotypes or the estimation of trait variability. Moreover, classical genetic linkage analysis is more powerful if performed in DZ twins. However, the twin method has some pitfalls, such as the necessity that collected samples be representative of both twin and general population. For this reason, over the last few years, a number of Countries have established population-based twin registers, which guarantee the maximum level of representation and, consequently, are of extreme value for epidemiological studies. Italy is also implementing a national twin register. The following is the description of the procedure that led to the establishment of the Italian Twin Registry. PMID- 14735843 TI - [Regression methods and causal inference: structural equations models]. AB - The estimate of correlations among observed outcomes is crucial in biomedical research, especially when the aim of the study is to infer, from the magnitude of these correlations, the causal influence of certain, sometimes latent, factors. In such situations, a typical regression approach, known as "structural equation models" (SEM), which was introduced in the 1970s, becomes significant. These models allow hypotheses to be formulated quite clearly, thanks to some explicit and rigorous graphical representations, on which the "path analysis" is based. SEM, which were initially used in economics, have in the past decade been applied in a wide variety of fields, especially in genetic epidemiology. It's in this field that SEM are extraordinarily effective, representing a simple yet powerful means of estimating the contribution of genes and the environment to the phenotypic expression of a given disease. To this end, data on twins are particularly useful, and in this case the correlation between the outcomes describes the extent of similarity of the twin phenotypes. From this standpoint, SEM undoubtedly constitute one of the most promising statistical tools for family studies and quantitative genetic research. The method can be easily extended to traditional epidemiology, and some interesting applications have already been developed in occupational and social epidemiology. In this paper, we describe in detail the SEM approach and discuss the use of these models in genetic epidemiology, using twin studies as an example. We also discuss the application of SEM in fields other than genetic research. PMID- 14735845 TI - [The "Tuscan Cigar" is American!]. PMID- 14735844 TI - [Role of epidemiology applied to a local area based on a study carried out in the community of Fiorano Modenese (ceramic district)]. AB - Mortality data in the province of Modena 1987-1996 show an excess for lung cancer in the municipality of Fiorano Modenese. In order to characterize the risk factors involved, a case-control study was carried out. Exposure to particulate matter was associated with lung cancer (OR 2.4 IC 95% 1.2-4.7). In the meantime, also as a consequence of this study, Fiorano Modenese Council started actions and obligations to control air pollutants emissions. The study is described as an example of good integration between local epidemiological knowledge and preventive interventions by Local Administration; problems and doubts surfacing from this experience are also pointed out. PMID- 14735846 TI - [Molecular epidemiology and patients' interests]. PMID- 14735847 TI - [A court for the rights of the disabled]. PMID- 14735848 TI - [A pseudo-epidemic of puerperal sepsis]. AB - Within a four-week period, five patients were admitted to the maternity ward of the Utrecht Children's Hospital diagnosed with puerperal sepsis due to group-A streptococcal infection. The clinical presentation was different for each patient. All patients recovered upon adequate antibiotic treatment. One of the children died, possibly due to sepsis and hypotension of his mother. As group-A streptococci can be extremely contagious and an epidemic was suspected, measures for additional hygiene were taken. Furthermore, all personnel at the maternity ward and the obstetric centre were tested. T-serotyping, M-genotyping, exotoxin A and C-gene amplification and pulsed field gel electrophoresis were used to characterize the cultured group-A streptococci. Cross-contamination was not found. Therefore, this increase in puerperal sepsis was attributed to polyclonal expansion rather than an epidemic. All mothers of newly born children who present with fever and lower abdominal pain should be suspected of group-A streptococcal infection. Evaluation and treatment in hospital is indicated due to a sometimes fulminant course. When group-A streptococci are cultured again in a new pregnancy, eradication therapy during pregnancy or prophylactic treatment during birth should be considered to prevent recurrent infection. PMID- 14735849 TI - [Clinical reasoning and decision making in practice. A man with pneumonia and ill smelling watery sputum: the truth revealed after 80 years]. AB - A 24-year-old man was admitted to hospital with pneumonia. On admission he was seen to have an asymmetrical build. During treatment of the lung infiltrate his clinical condition deteriorated. On the third day he coughed up great quantities of fluid which had a urine-like smell. The concentration of creatinine in this fluid was the same as in urine. On X-ray of the thorax, a massive accumulation of pleural fluid was seen. Shortly after aspiration of 1000 ml of pleural fluid the patient died. At autopsy, an ectopic kidney was found in the left thoracic cavity. The pneumonia had caused an abscess that had broken into the pelvis of this ectopic kidney causing the loss of urine into the pleural cavity (urothorax) and 'uroptysis'. On the basis of anatomical and embryological aspects it is debatable if this case was genuine. It is in fact a duplication of a case report published in this journal in 1923 the reliability of which was never clarified. Biographical information from Professor A. Querido (1901-1983) which has since become available indicates that the case was faked by mischievous medical students preparing for their examinations. They had never imagined that the editors might actually accept it for publication. The case report of 1923 has now been retracted. PMID- 14735850 TI - [Cosmetic surgery, certificates and the 'head-baker' of Eeclo]. AB - The legend of the head-baker of Eeclo, an imaginary cosmetic surgeon, clearly illustrates that the improvement of the human physionomy was of great interest to people as long ago as the mid-16th century. In order to display their credentials the surgeons of that time and in the succeeding centuries would display certificates that testified in glowing terms to their knowledge and expertise. One only has to open a modern newspaper to see that this phenomenon has not died out. PMID- 14735851 TI - [The history of epilepsy in the Dutch Journal of Medicine]. AB - Epilepsy has always been a mysterious condition. Between 1857-2002 the Dutch Journal of Medicine has devoted countless articles to this condition. At the end of the 19th and the beginning of the 20th century epilepsy was associated with a variety of psychological disorders and a high rate of inheritability. Many theories as to its causes were advanced. During the last century the development of new diagnostic techniques led to rapid changes in reasoning. The development of electroencephalography had a particularly big effect. It quickly became possible to distinguish different types of epilepsy according to their causes and clinical manifestations. Epilepsy could also be distinguished from other conditions better than previously. Types of treatment varied according to the current opinion on its causes. In most patients the best results are obtained through drug treatment; for a few, surgery is necessary. PMID- 14735852 TI - [Leopold Meyler (1903-1973): a pioneer in the field of side affects of drugs]. AB - In 1951, more than a decade before the first rumblings of the thalidomide catastrophe, a remarkable book appeared in the Netherlands under the authorship of Leopold Meyler. It appeared a year later in an English translation, as Side effects of drugs. This was a remarkable book at a time that medical interest in drugs--and particularly new drugs--was limited almost entirely to their proven or possible therapeutic benefits. Meyler was a specialist in internal medicine in Groningen. He was Jewish and was forced to go into hiding during World War II. He suffered a great deal during this time and after the war he was admitted to a sanatorium where he was treated for tuberculosis. It was here that he was advised to start the systematic collection of adverse-drug reactions reported in literature. Initially there was quite a lot of criticism of his work but it gradually gained recognition and he was appointed the first professor of Clinical Pharmacology at Groningen University in 1968. In his own time his was a lone voice crying in the wilderness. Had his voice been heeded earlier, the extent of the thalidomide disaster might well have been much more limited. His work still continues to contribute to the safe use of drugs. PMID- 14735853 TI - [The haematogenous reproduction theory of Aristotle]. AB - From the very beginning, man has been fascinated by the continuous coming into being of new life on Earth. Archeological and anthropological data indicate that in prehistoric times, reproduction was attributed to a fertility goddess that required no sperm for this purpose. In the early historical Middle East it was believed that a godly being brought about pregnancy by using male and female semen. It was the merit of the Greek philosophers of the 6th-3rd century B.C. to realize that reproduction was governed by natural laws. Several theories were developed to understand how reproduction could occur. The haematogenous theory of reproduction, developed by Aristotle, has received the most attention. The essence of this theory is that the male sperm, with a haematogenous origin, causes the development of an embryo from menstrual blood present in the female uterus. The theory survived about 2000 years, with modifications, and was also introduced into Christianity. It was only about 150 years ago that the reproduction theory based on hypotheses was changed into a reproduction science based on facts. PMID- 14735854 TI - [Chlorosis, the lost disease of languid young women]. AB - Chlorosis or 'green sickness' was frequently seen in languid girls and young women in the 19th century but disappeared completely in the first part of the 20th century. The clinical picture comprised menstrual disorders such as ameonrrhoea, pallor and many vague symptoms including apathy and hypochondria. At a later stage anaemia and iron deficiency became prominent characteristics. The skin was reported to take on a greenish hue, but this is disputable. Related diseases were hysteria and anorexia. In the middle of the 19th century hydrotherapy was treatment of choice, and later on iron therapy came to the fore. In 1898 Catharine van Tussenbroek, the first female Dutch gynaecologist, pointed to the social factors at the root of the disease: the lack of perspective for young girls in society at that time. The disappearance of the disease can be partially attributed to improved diagnostics but more so to changes in the social position of women around the turn of the century. PMID- 14735855 TI - [What do you actually see? Visual impairments and their simulation for well seeing subjects]. AB - Most physicians know little about how persons with limited vision perceive something. Sometimes, simple technical aids can make it possible to get an idea as to what a visual disorder means to the person involved. Examples of this are the wearing of glasses of varying positive power and light transmittance, or the study of case histories, paintings and photographic simulations. Among the latter are a woman who painted what she 'saw' with her removed eye, an anophthalmic man who painted from memory, and the painting of a protanopic artist who chose colours by reading the labels on the paint tubes. Thus one may gain insight in disturbances such as phantom images, diminished visual acuity and visual field loss, metamorphopsia, diplopia, dazzling, and visual agnosia. And also in disturbed image fusion, in depth and stereoscopic vision, dark adaptation and colour vision, as well as diminishing visual functions at high age. This article contains colour illustrations that simulate the disturbances. PMID- 14735856 TI - [Hereditary hyperferritinaemia-cataract syndrome]. PMID- 14735857 TI - [Duloxetine and other tricyclic antidepressants: pharmacodynamic effects in the lower urinary tract]. AB - INTRODUCTION: During years the pharmacology of the lower urinary tract function has been presided by the protagonism of the autonomic nervous system and its components, sympathetic and parasympathetic. Recent investigations proved a greater complexity of micturional dynamics, attributing a more important role to the central nervous system (SNC), to the striated sphincter and to the afferent arc, offering a new opportunity to the Tricyclic Antidepressants (T.A.), reason for this general review. MATERIAL AND METHODS: Using as reference the work of the First Consultation on Incontinence (Montecarlo, 1999), the previous and later publications about T.A. have been reviewed, including experimental (isometric and "in vivo" studies) and clinical studies, investigating on pharmacological evidences, mechanism of action, tolerance and other effects of T.A. RESULTS: Only a reduced group of T.A. have been submitted to experimental evaluations and employed in clinical trials. The recent works on Duloxetine have waked up a special interest by their pharmacological potential. CONCLUSIONS: New knowledge on the peripheral and central control of the continence-miction dynamic offer new pathways for the treatment with T.A., whose effectiveness and tolerance are reviewed. PMID- 14735858 TI - [Current multidisciplinary treatment of metastatic prostatic cancer]. AB - Prostate cancer is one of the commonest causes of cancer-related death in the western world. The morbi-mortality associated is usually a direct consequence of metastatic spread to bone, in up to 50% of patients at first presentation. The aim of treatment of metastatic patients is to alleviate and to prevent the distressing symptoms. The approach include hormone-therapy, radiotherapy, radio nuclides, surgery, chemotherapy, bisphosphonates and new drugs (agents that inhibit angiogenesis, immunotherapy and therapies that affect the differentiation). Decisions about therapy must also take into consideration the androgen-dependent or independent, so hormone-therapy is the first step of the treatment; the number and location of bone metastases; the severity of symptoms; the available of therapies; the status performance of patient; the prognosis and the cost-effect relationship. Some treatments have established indications whereas others are still in process of study in order to determinate their efficacy, their model of treatment and their indications. This article revises and updates these treatments. PMID- 14735859 TI - [Microbial sensitivity of Escherichia coli in community-acquired urinary tract infections]. AB - OBJECTIVE: For effective empiric therapy of urinary tract infections in the extra hospital setting the susceptibility pattern of uropathogens should be considered. Moreover, the evolution in sensitivity can be observed when comparing with susceptibility patterns in the previous years. This paper presents an analysis of our experience with Escherichia coli. MATERIAL AND METHODS: During 2002 and 1998, 895 and 595 strains of Escherichia coli respectively, isolated from extrahospitalary bacteriurias were collected in ten health centers in Bierzo (Leon, Spain). Sensitivity to nine most commonly antibiotics used in the clinical practise was determined. The existence of significant differences of susceptibility among years (2002-1998) was analyzed by the chi square test. RESULTS: Escherichia coli accounted for 63.4% of all isolates in 2002 and 50.8% in 1998. The prevalence of in-vitro susceptibilities to antibiotics were (2002 1998): fosfomycin (99.2%-99.3%; p = NS*), cefixime (98.3%-92.9%; p < 0.001), cefuroxime (96.5%-94.1%; p < 0.05), nitrofurantoin (94.5%-86.9%; p < 0.001), amoxycillin-clavulanic acid (93.1%-90.1%; p < 0.05), ciprofloxacin (77.1%-81.6%; p < 0.05), norfloxacin (75.8%-80.3%; p < 0.05), cotrimoxazole (71.5%-73.4%; p = NS*) and ampicillin (44%-41.4%; p = NS*). (*NS = No significant differences). CONCLUSION: The knowledge of the sensitivity of uropathogens to antimicrobians in a specific medium can allow us to use antibiotics rationally and initiate empirical therapy. PMID- 14735860 TI - [Relationship between serum testosterone levels and prostatic cancer]. AB - INTRODUCTION: The association between testosterone and prostate cancer badly is understood. The testosterone levels are not conclusive to distinguish benign prostate processes of malignant. The objective of our work is to determine the relation between levels of serum testosterone and prostate cancer. MATERIAL AND METHOD: 38 patients with prostate cancer and a control group of 32 were studied in prospective form, with ages that fluctuated between the 50 and 80 years. The total and free testosterone was determined by radio-immunoensayo. The statistical analysis was descriptive inferential, with bands of 95% confidence. RESULTS: The average values of total and free testosterone, were significantly smaller in patients with prostate cancer. Any correlation between gleason, prostate antigen and stage with testosterone, was not demonstrated. CONCLUSIONS: Our work demonstrated that the testosterone is significantly smaller in the patients with prostate cancer that in the controls. PMID- 14735861 TI - [Use of intrarectal lidocaine gel in ultrasound-guided transrectal biopsies of the prostate]. AB - OBJECTIVE: To know in a quantitative manner the degree of discomfort and pain of the biopsies of the prostate and to evaluate the effectiveness of the transrectal lidocaine. MATERIAL AND METHOD: We performed 140 transrectal biopsies of the prostate, Patients were included on a random basis into two arms: one of them received intrarectal lidocaine, 20 mg (group 1, n = 71) and the other group received placebo (group 2, n = 28) both of them ten minutes prior the proceeding. RESULTS: The global pain mean was 3.7 (0 no pain, 10 highest pain) and the global discomfort mean was 3.5. The group 1 patients showed a trend to feel less pain and discomfort although it did not reach the necessary statistic significance (p = 0.7 y p = 0.5 respectively). CONCLUSIONS: We do not achieve the good results obtained by other groups in order to decrease the degree of pain and discomfort with the use of intrarectal lidocaine. We did not find relationship between the PSA level, previous biopsies, intrarectal lidocaina and degree of information received and the degree of pain and discomfort. PMID- 14735862 TI - [Penile cancer. Review of 18 cases]. AB - INTRODUCTION: Squamous cell carcinoma is the most common tumor of the penis. Nodal metastases are relatively common, but distant disemmination is very rare. MATERIAL AND METHODS: From 1990 to 2002, we diagnosed and treated 18 cases of carcinoma of the penis. The minimum follow-up was 2 years (range 2-11 years; mean 49 months). Treatment of the primary lesion is usually by partial amputation of the penis, which enables us to determine the pathological stage and the histological grade of the tumor and, consequently, our approach to the regional lymph nodes. RESULTS: Palpable inguinal nodes after antibiotherapy remained in six out of seven patients. Inguinal lymphadenectomy was performed early in 4 cases. Postoperative complications were present in the 4 cases, lymphedema being the most frequent one (100%). CONCLUSIONS: The T and the histological grade of the primary lesion must be considered when deciding the approach in the management of the lymph nodes as unneccesary lymphadenectomy can be avoided and those at high risk of lymph node invasion can be treated radically and timely. PMID- 14735864 TI - [Nephroblastomatosis: which therapeutic approach should be used? Report of 2 cases]. AB - Nephroblastomatosis is a complex of pathological conditions defined by the persistence of nephrogenic elements after the conclusion of nephrogenesis (beyond week 36 of gestation) that conserve the capacity to evolve into nephroblastoma. Luckily, this malignant transformation is exceptional (< 1%(1)), which is why the disease is considered to have a good prognosis. The nephrogenic remains are frequently associated with beckwith-Wiedemann syndrome, hemihypertrophy and aniridia, in which case there is a greater risk of the development of Wilms tumor. Consequently, these children must be followed-up closely. The difficulty lay in knowing when to act, which children should be treated, and what treatment is appropriate because what little has been published in the literature is debatable. We report the cases of two patients, one with unilateral nephroblastomatosis and the other bilateral, who required surgical treatment after not responding to chemotherapy; and we expose our therapeutic algorithm and follow-up strategy. PMID- 14735863 TI - [Treatment with the Newbit technique in patients with Peyronie's disease]. AB - OBJECTIVE: Evaluate the results of Nesbit's technique in patients with Peyronie's disease. METHODS AND MATERIALS: Between 1990 and 2002, 45 patients were treated using the Nesbit's procedure to correct Peyronie's disease curvature. Mean age was 57.6 (range 41-73). Dorso lateral incurvation was the most common. RESULTS: Total correction of the curvature in 40 patients (88%). High grade of satisfaction in 39 patients (86.66%). CONCLUSION: Nesbit's operation is an easy and effective surgical technique in the peyronie's disease for curvature correction. PMID- 14735865 TI - [Female urethral diverticulum. Report of a case]. AB - PURPOSE: We describe the successful repair of a large and complex urethral diverticulum in a female by transvaginal approach. Epidemiology, diagnostic methods, treatments and complications of female urethral diverticula are reviewed. PATIENTS AND METHODS: A 35-year-old woman with a history of postvoid dribbling, dyspareunia and recurrent urinary tract infections for 4 months was referred. Magnetic resonance imaging demonstrated two fluido-filled collections in the pelvis of 3.5 and 1 cm in size respectively which may be a very large and complex diverticulum, however, Bartholin gland cyst could not be rule out. Cystourethroscopy revealed a urethral diverticulum at 10 mm from the bladder neck with two ostia. It was performed transvaginal diverticulectomy and an anterior vaginal wall flap was placed. The published literature on female urethral diverticula was identified using a Pubmed Medline search and analysed. RESULTS: Convalescence was unremarkable. Suprapubic cystostomy tube was removed 2 weeks after surgery. The patient regained normal voiding. In the published literature there are no agreement neither in the diagnostic nor in the surgical techniques for female urethral diverticula. CONCLUSIONS: Urethral diverticula are diagnosed with increasing frequency. However, this entity continues to be overlooked because the symptoms may mimic other disorders. Cystourethroscopy, retrograde urethrograme using a double balloon catheter and recently magnetic resonance imaging may diagnose this disease. The cure rate of urethral diverticula with appropriate surgical management has a range of 86-100%. Complete excision through the anterior vaginal wall is the most successful treatment modality with minimum postoperative complications. PMID- 14735866 TI - [Scrotal leiomyoma: report of a case]. AB - Scrotal leiomyoma is a benign rare tumour, asymptomatic, which origin is the dartos muscle and the election treatment is surgical. We report a new case in a 68-year-old patients with a 10 years history of a scrotal tumour and anatomopathological diagnostic post surgery was leiomyoma. PMID- 14735867 TI - [Localized amyloidosis of the seminal vesicles]. AB - Amyloidosis of the seminal vesicles is a common finding in autopsies, with increased incidence in older population. It is usually asymptomatic. We report a case of symptomatic localized amyloidosis of the seminal vesicles, with hemospermia and suprapubic pain. Diagnosis was achieved through ultrasound-guided transrectal biopsy. Systemic amyloidosis must be ruled out through proper evaluation. Seminal vesicle enlargement secondary to amyloid deposit may be misdiagnosed as carcinomatous invasion. PMID- 14735868 TI - [Late recurrence of penile epidermoid carcinoma]. AB - OBJECTIVE: We report a recidive of penis carcinoma after 21 years of surgery. METHODS: This is the case of a male of 85 years old, with partial penectomy; by epidermoid carcinoma. In physics exploration, that it displays excrecente lesion at level of gland of 1 month of evolution. Biopsy being the result of epidermoid carcinoma reason why is made. Total penectomy, a with cutaneus uretrostomy. RESULTS: The histopathological study of the piece, confirm a epidermoid carcinoma well differentiated. PMID- 14735869 TI - [Solitary fibrous tumor. Two additional cases with urologic implications]. AB - Solitary fibrous tumour is a well defined pathological entity originally described as a tumour of the pleura, but the occurrence of this neoplasm has increasingly been described at other sites. At present the development of these tumours is recognized as possible in virtually all anatomical sites. The rarity of this type of pathology has not allowed up to the present time clarification of the histogenesis, clinical behaviour, treatment and prognosis of these tumours. Differential diagnosis becomes important, especially in extra-thoracic sites so as not to mistake them for much more aggressive mesenquimal tumours. Two additional cases of this neoplasm are presented, one of them responsible for obstructive uropathy, accidentally found in a patient suffering renal trauma. The second tumour was located in the spermatic cord. A review of the theme is presented based on the available literature. PMID- 14735870 TI - [Usefulness of PET in the diagnosis and response to treatment of extragonadal germinal tumor with atypical presentation]. AB - Primary tumors of extragonadal origin are rare, with fewer than 1000 cases described in the literature. Although the exact incidence of EGTs is unknown, clinical data suggest that roughly 3% to 5% of all germ cell tumors. We expose a case report of EGT with unusually clinic presentation. We present our diagnostic and therapeutic experience in this injuries. PMID- 14735871 TI - The gray areas of boundary crossings and violations. AB - The term "boundary violation" has become synonymous with unethical practice in psychotherapy, prompting a "black-or-white" view among clinicians and boards of review. But the current conceptual ambiguity about boundary interventions subjects clinicians to after-the-fact second-guessing that can be professionally ruinous at worst and may contribute to stultifying defensive therapeutic rigidity at best. It is crucial to demarcate the differences between boundary violations and boundary crossings (to be defined below) as clearly as possible, to describe the "gray areas" of each, and to recognize the heterogeneity of boundary violations and boundary crossings. These matters are of additional importance to those who teach, train, and supervise the next generation of clinicians. This paper will attempt to delineate a more clinically realistic and useful view of boundary crossings than has been proposed before, and illustrate the practical advantages of recognizing the spectrum of boundary interventions. PMID- 14735872 TI - Psychotherapy of Arab patients in the West: uniqueness, empathy, and "otherness". AB - The study of Arab patients seeking treatment for their psychological problems in the West has previously been underrepresented in mainstream American journals. Notwithstanding various attempts that deal with Arab Americans as a minority group, there has been a paucity of scholarship dealing with Arab patients' unique characteristics related to sociopolitical, cultural, and other factors that impact the therapeutic process for those individuals who are not acculturated to the American way of life. These patients present challenges to their therapists owing to the contrasting cultural understanding and conceptualization of mental illness and therapeutic process. Therapists need to fully appreciate the relationship between culture and psychotherapy. Patients' and clinicians' awareness of differences may contribute to the ability of both sides of the therapeutic dyad to overcome some of the differences encountered when Arab patients are treated outside their cultural domain. A case vignette serves to illuminate how issues of cultural transference and countertransference can be managed for the benefit of the patient and the enlightenment of the therapist. PMID- 14735873 TI - Differential-diagnosis of obsessive-compulsive symptoms in the borderline personality disorder. AB - Obsessive-compulsive symptoms are clinically unspecific and can be found in numerous disorders. Within the framework of psychotherapeutically treatable illnesses, the classical obsessive-compulsive neurosis can be distinguished from early anancastia in the case of borderline personality disorder. The text refers to some aspects of the obsessive-compulsive disorder within these earlier disorders. At least five characteristics can be discussed: In the course of this, the varying functions of obsessive-compulsive symptoms for the inner psychological organization become clear and specifically show that on a low structural level, symptoms do not appear to be primarily pathological but are a part of a "quasi-physiological" and ego-sustaining mechanism. What remains open to discussion in the end is, whether there may be an important substratum for the basis of a historiographical biology as called repeatedly for from psychosomatic theory repeatedly. In addition to this, the awareness of both forms is an important requirement for treatment. PMID- 14735874 TI - Emotions: a relational view and its clinical applications. AB - In this paper, three major tasks are undertaken: 1. showing that our traditional understanding of the nature of emotions, which equates them with certain sorts of inherently private affective experiences that are brought about by various causal factors (esp., exciting events, cognitive interpretations, and biological states of affairs), does justice neither to the conceptual nor to the empirical facts; 2. presenting an alternative conception of emotions as a specific class of perceived relationships between oneself and some person, object, event, or state of affairs; 3. demonstrating how this relational conception of emotions heuristically suggests a far greater range of therapeutic options than does the traditional view. PMID- 14735875 TI - An existentialist approach to anorexia nervosa. AB - Anorexia nervosa is an attempt of the self to refuse itself and devise an other self, with dissolution of the link between the self and the body. Although anorectics are often described as academically capable and attractive, in the author's experience they were preoccupied with superficial issues, their drive came not from the seductive promise of intellectual stimuli, but from negative expectations that needs cannot be met, driving one to rise above/outside of needs to "not need." The new self, incorporeal and totally controlled, in its controlling part, is at times perceived as a separate it controlling the mind. The new self, that so much wanted to be in control, is out of control by being under the orders of the it; supposedly not greedy, it has found new object of greediness in weight loss. The anorectic's feeling of "superiority" has relevant therapeutic implications. PMID- 14735876 TI - Child-survivors of the Holocaust: age-specific traumatization and the consequences for therapy. AB - Keilson was the first who differentiated between various age groups, regarding the relationship between stressful Holocaust situations and their ultimate effect on later life. In this paper I will compare the developmental needs of different age groups and the devastating reality child-survivors had to go through. In the therapeutic situation with child-survivors who are now in their sixties and seventies, different defense styles become apparent. The therapist will be confronted with an adult who herds an abandoned child within, who has fragmented or no memories of the past and who is not used to being in touch or revealing his hidden painful feelings of longing and grief. PMID- 14735877 TI - Family approach with grandchildren of Holocaust survivors. AB - Although the transgenerational transmission of Holocaust trauma is now well documented, this subject remains a source of considerable controversy. Moreover, the literature regarding the grandchildren of Holocaust survivors (GHSs, the third generation) is much sparser. We present here several clinical observations, that we made during therapy sessions with certain families of Holocaust survivors (HSs). These families consulted with us because of the symptoms presented by the GHSs as adolescents. These families were characterized by some specific patterns in their relationships that led us to consider that the symptoms of the third generation might be a consequence of the transgenerational transmission of Holocaust trauma. We also describe the clinical strategy we developed to assist these families of HSs. This strategy consisted of an attempt to reinforce the relationships between GHSs and their grandparents, the Holocaust survivors. PMID- 14735878 TI - Working with metaphor. AB - This article explores and affirms the importance of metaphor, in both conceptual and poetic thought, and its relevance to the practice of psychotherapy. It examines some of the obstacles commonly encountered as we attempt to become more aware of, and responsive to, the metaphors used by our clients. It continues by describing a simple six-stage model for working with client-generated metaphors, and concludes with a case example demonstrating the effectiveness of metaphor exploration in clinical work. PMID- 14735879 TI - Narcissism and spirituality in Flannery O'Connor's stories. AB - Virtually any short story by Flannery O'Connor could serve as a poignant case study of narcissism. While narcissism in the guises of ambition and hubris is an ancient literary theme, O'Connor's protagonists vividly exemplify a syndrome of covert and hypervigilant narcissism that has been well characterized in the literature. Her work also strongly implies that narcissism and spirituality (particularly Christianity) are antithetical, and two of her stories--"The Enduring Chill" and "The Lame Shall Enter First"--are analyzed with respect to this belief. The characters of Asbury and Sheppard exhibit classic narcissistic signs and symptoms as delineated by Kohut and Kernberg. The complex relationship of narcissism to evil, spirituality, and contemporary culture is explored, and it is argued that narcissism has a prominent spiritual dimension that raises questions about the role of values in psychotherapy. PMID- 14735880 TI - The $1000 genome: ethical and legal issues in whole genome sequencing of individuals. AB - Progress in gene sequencing could make rapid whole genome sequencing of individuals affordable to millions of persons and useful for many purposes in a future era of genomic medicine. Using the idea of $1000 genome as a focus, this article reviews the main technical, ethical, and legal issues that must be resolved to make mass genotyping of individuals cost-effective and ethically effective. It presents the case for individual ownership of a person's genome and its formation, and shows the implications of that position for rights to informed consent and privacy over sequencing, testing, and disclosing genomic information about identifiable individuals. Legal recognition of a person's right to control his or her genome and the information that it contains is essential for further progress in applying genomic discoveries to human lives. PMID- 14735881 TI - Ethical guidelines for use of electronic mail between patients and physicians. AB - This report examines the ethical implications of electronic communication, focusing on the use of electronic mail (e-mail), considers its impact on a previously established patient-physician relationship, and the limitations in using e-mail to create a new patient-physician relationship. In its recommendations, this report offers guidance to physicians who use electronic mail to communicate with patients and online users. These guidelines maintain that e-mail should not be used to establish a patient-physician relationship, but rather to supplement personal encounters. When using e-mail, physicians hold the same ethical responsibilities to their patients as they do during other encounters and that information must be presented in a manner that meets professional standards. The report requires that physicians notify patients of e mail's inherent limitations and that patients be given the opportunity to accept these limitations prior to the communication of privileged information. Finally, physicians should be aware of privacy and confidentiality concerns when using e mail to communicate with patients. PMID- 14735882 TI - Use of health-related online sites. AB - This report offers recommendations to physicians who provide information or services through online sites. The recommendations maintain that physicians responsible for health-related information should ensure that it is accurate, timely, reliable, and scientifically sound. Also, advice to the online users with whom physicians do not have pre-existing relationships or the use of decision support programs that generate personalized information directly transmitted to users should be consistent with general and specialty-specific standards. In particular, these standards address truthfulness, protection of privacy, informed consent, and disclosures including limitations inherent in the technology. Finally, physicians who establish or are involved in health-related online sites must minimize conflicts of interest and commercial biases and, if patient specific information is transmitted, they must provide high-level security protections, as well as privacy and confidentiality safeguards. PMID- 14735883 TI - Remember Saddam's human guinea pigs. PMID- 14735884 TI - A response to "Children in clinical research: a conflict of moral values" by Vera Hassner Sharav. PMID- 14735885 TI - A response to "Children in clinical research: a conflict of moral values" by Vera Hassner Sharav. PMID- 14735886 TI - Creative supplement: The cancer patient experience-New poetry and prose. PMID- 14735887 TI - A response to commentators on "All gifts large and small". PMID- 14735888 TI - A response to "Crossing species boundaries" by Jason Scott Robert and Francoise Baylis. PMID- 14735890 TI - A new concept of three-dimensional endoscope for endoscopic surgery. PMID- 14735889 TI - A response to commentators on "Crossing species boundaries". PMID- 14735891 TI - Status and future directions: RadioGraphics and RSNA Education 2004. PMID- 14735892 TI - [Adequate dose--that is the solution. Discussion of article by J. Spacil: Maximal or medium drug doses in cardiovascular diseases?]. PMID- 14735893 TI - [Not Available]. PMID- 14735894 TI - Time to start over. PMID- 14735902 TI - A prejudiced patient. PMID- 14735904 TI - [The HIV escort]. PMID- 14735903 TI - Frequent activation of AKT2 kinase in human pancreatic carcinomas. AB - Activation of AKT/protein kinase B promotes a variety of biological activities important in tumorigenesis, such as cell survival and cell cycle progression. We previously demonstrated amplification and overexpression of the AKT2 gene in a subset of human pancreatic carcinomas. In this investigation, we assessed AKT2 catalytic activity in 50 frozen pancreatic tissues (37 carcinomas, four benign tumors and nine normal pancreata) by in vitro kinase assay. Twelve of 37 (32%) pancreatic carcinomas showed markedly elevated levels of AKT2 activity compared to normal pancreata and begin pancreatic tumors. To delineate mechanisms contributing to AKT2 activation in malignant pancreatic tumors, we examined the status of upstream components of the phosphatilydlinositol 3-kinase (PI3K)/AKT pathway. Western blot analysis revealed loss of PTEN protein expression in two of the 12 pancreatic carcinomas with activated AKT2. In vitro PI3K assays demonstrated high levels of PI3K activity in seven carcinoma specimens that showed AKT2 activation. Immunohistochemical staining confirmed high levels of phosphorylated (active) AKT in malignant pancreatic tumors compared to normal pancreata. Overall, these data suggest that upstream perturbations of the PI3K/AKT pathway contribute to frequent activation of AKT2 in pancreatic cancer, which may contribute to the pathogenesis of this highly aggressive form of human malignancy. PMID- 14735905 TI - A multicenter, randomized, double-blind, controlled trial of nebulized epinephrine in infants with acute bronciolitis. PMID- 14735906 TI - Evaluation of impermeable covers for bedding in patients with allergic rhinitis. PMID- 14735907 TI - Effect of viewing smoking in movies on adolescent smoking initiation: a cohor study. PMID- 14735908 TI - Efficacy of sertraline in the treatment of children and adolescents with major depressive disorder: two randomized controlled trials. PMID- 14735909 TI - Chromosome mapping in lactic acid bacteria. AB - The chromosome structure of lactic acid bacteria has been investigated only recently. The development of pulsed-field gel electrophoresis (PFGE) combined with other DNA-based techniques enables whole-genome analysis of any bacterium, and has allowed rapid progress to be made in the knowledge of the lactic acid bacteria genome. Lactic acid bacteria possess one of the smallest eubacterial chromosomes. Depending on the species, the genome sizes range from 1.1 to 2.6 Mb. Combined physical and genetic maps of several species are already available or close to being achieved. Knowledge of the genomic structure of these organisms will serve as a basis for future genetic studies. Macrorestriction fingerprinting by PFGE is already one of the major tools for strain differentiation, identification of individual strains, and the detection of strain lineages. The genome data resulting from these studies will be of general application strain improvement. PMID- 14735910 TI - Endocrine-related resources from the National Institutes of Health. PMID- 14735911 TI - Isolation and characterization of a fucoidan-degrading marine bacterium. AB - Fucoidan, a mixture of sulfated fucose-containing polysaccharides, was prepared from the algal bodies of Cladosiphon okamuranus (class Phaeophyceae, order Chordariales, family Chordariaceae) with a yield of 2.0% of the wet weight of the alga. To obtain enzymes that digest the fucoidan, we screened bacteria in the guy contents of the sea cucumber Stichopus japonicus for their ability to decrease the fucoidan in their culture media, and successfully isolated one bacterial strain that could decrease it. The bacterial strain was gram-negative and possessed menaquinone 7 as the predominant respiratory quinone, and the GC content of its genomic DNA was 52%. The results of the phylogenetic analysis of its 16S ribosomal DNA sequence indicated that the bacterial strain was a member of the division "Verrucomicrobia." However, as the bacterial strain is phylogenetically and phenotypically distinct from verrucomicrobial species described previously, the strain was assumed to be a new member of the division "Verrucomicrobia." When the bacterial strain was cultivated in an algal fucoidan containing medium, the strain decreased fucoidan from C. okamuranus (44%), Nemacystus decipiens (19%), Laminaria japonica (31%), Kjellmaniella crassifolia (23%), sporophyl of Undaria pinnatifida (22%), Fucus vesiculosus (42%), and Ascophyllum nodosum (61%). PMID- 14735912 TI - A silent antipode: The making and breaking of psychoanalyst Wilhelm Stekel. AB - Wilhelm Stekel, one of Freud's earliest followers , was expelled from the psychoanalytic movement in 1912 ostensibly because he did not know how to behave himself. Although he remained active as a psychoanalyst, his post-1912 work was mostly neglected, and consequently his historic import is seriously undervalued. The author reviews recent literature, reexamines the Freud-Stekel break, and focuses on Stekel's role as silent antipode. Freud's reference to an unnamed individual in his 1907 Gradiva paper (S. Freud, 1907/1959b) - commonly believed to be Jung - is now identified as Stekel. This not - unimportant correction of the historical record begins the exploration of a hitherto-undocumented antagonistic dialogue between Stekel and Freud. PMID- 14735913 TI - B.F. Skinner and the auditory inkblot: The rise and fall of the verbal summator as a projective technique. AB - Behaviorist B.F. Skinner is not typically associated with the fields of personality assessment or projective testing. However, early in his career Skinner developed an instrument he named the verbal summator, which, at one point, he referred to as a device for "snaring out complexes," much like an auditory analogue of the Rorschach inkblots. Skinner's interest in the projective potential of his technique was relatively short lived, but whereas he used the verbal summator to generate experimental data for his theory of verbal behavior, several other clinicians and researchers exploited this potential and adapted the verbal summator technique for both research and applied purposes. The idea of an auditory inkblot struck many as a useful innovation, and the verbal summator spawned the tautophone test, the auditory apperception test, and the Azzageddi test, among others. This article traces the origin, development, and eventual demise of the verbal summator as an auditory projective technique. PMID- 14735914 TI - The adoption history project: A new on-line resource. AB - The author offers a brief description of The Adoption History Project, a Web site launched on June 1, 2003, that covers the people and organizations, issues, and studies, that shaped adoption in theory and practice throughout the 20th century. The Adoption History Project offers many brief biographical and topical sketches, a wealth of primary texts (excerpted from published and unpublished sources), numerous images, a timeline, bibliography, and a search engine. Historians of psychology will be especially interested in how the consolidation of professional communities (in psychology, social work, and medicine) influenced family formation and in the history of the adoption research industry. PMID- 14735915 TI - Isolation and characterization of novel Helicobacter spp. from the gastric mucosa of harp seals Phoca groenlandica. AB - Since the recent discovery of Helicobacter cetorum in cetaceans and its role in the development of gastritis, speculation has existed as to whether pinnipeds have Helicobacter spp. associated gastritis and peptic ulcer disease. The gastric mucosa of 4 stranded harp seals Phoca groenlandica from the Massachusetts coastline were assessed for Helicobacter spp. by culture and PCR. We cultured 2 novel Helicobacter spp. from the pyloric antrum of 1 of the 4 harp seals studied, and identified these by PCR in 2 of the 4 seals. Both gram-negative bacterial isolates were catalase- and oxidase-positive. However, a fusiform helicobacter with flexispira morphology was urease-positive, and a spiral-shaped helicobacter was urease-negative. Slender, spiral and fusiform-shaped bacteria were detected in the gastric mucosa by the Warthin-Starry stain. Histopathologic analysis revealed mild diffuse lymphoplasmacytic gastritis within the superficial mucosa of the pyloric antrum of both infected seals. The 2 bacterial isolates were classified by 16S rRNA analysis; they clustered with other enteric helicobacters and represent 2 novel Helicobacter spp. The urease-negative bacterial isolate clustered with H. canis and the urease-positive isolate clustered with an isolate from a sea lion and isolates from sea otters. This cluster of pinniped isolates has 97 % similarity to a number of Helicobacter species, but appears to be most closely related to other helicobacters with flexispira morphology. These findings suggest that the novel Helicobacter spp. may play a role in the etiopathogenesis of gastrointestinal diseases in pinnipeds. To our knowledge, this represents the first isolation and characterization of a novel Helicobacter spp. from pinnipeds. PMID- 14735916 TI - Occurrence, histopathology and experimental transmission of hepatopancreatic parvovirus infection in Penaeus monodon postlarvae. AB - Hepatopancreatic parvovirus (HPV) was detected in samples of Penaeus monodon post larvae (PL-13, PL-18, PL-19, PL-26) from 2 hatcheries in 2 provinces (Samar and Iloilo) in the Philippines. The percentage of infection was 20 to 100% in postlarvae obtained from the hatchery in Samar in August 2001. Postlarvae from the hatchery in Iloilo, sampled in October and November 2001, had 70 to 99% HPV infection. Wet mounts of squashed hepatopancreatic tissue stained with malachite green (wet-mount technique) and histopathology revealed the presence of large, usually single, basophilic intranuclear inclusion bodies in the distal tubules, which led to displacement of the nucleoli. Light microscopy showed ovoid to spherical inclusion bodies, 5 to 11 mmicrom in diameter. Transmission electron microscopy revealed that the inclusion bodies were composed of electron-dense granular material and virions. The virions appeared roughly spherical and averaged 18 to 22 nm in diameter. An experiment was undertaken to induce HPV infection by feeding P. monodon postlarvae with virus-infected postlarvae. P. monodon postlarvae (PL-16), initially determined as free from HPV, were found HPV positive 24 h after being fed with infected material. The percentage of infection ranged from 30% at Day 1 post-infection (p.i.) to 100% at Day 7 p.i. determined by the wet-mount technique and by histopathology. This is the first report of a successful horizontal transmission of HPV in P. monodon postlarvae. PMID- 14735917 TI - Serological relationships among genotypic variants of betanodavirus. AB - Betanodaviruses, the causative agents of viral nervous necrosis or viral encephalopathy and retinopathy, are divided into 4 genotypes based on the coat protein gene (RNA2). In the present study, serological relationships among betanodavirus genotypic variants were examined by virus neutralization tests using rabbit antisera raised against purified virions of strains representative of each genotype. All 20 isolates examined shared epitopes for neutralizing, but they fell into 3 major serotypes (A, B, C). This sero-grouping is in part consistent with their genotypes, i.e. Serotype A for striped jack nervous necrosis virus (SJNNV) genotype, Serotype B for tiger puffer nervous necrosis virus (TPNNV) genotype, and Serotype C for both redspotted grouper nervous necrosis virus (RGNNV) and barfin flounder nervous necrosis virus (BFNNV) genotypes. The serological relatedness between RGNNV and BFNNV genotypes may result from their relatively higher similarity in RNA2 sequences. In neutralization tests using antisera of kelp grouper Epinephelus moara, which were raised against recombinant coat proteins representing each genotype, anti-SJNNV and anti-TPNNV sera neutralized only the homologous strain, and anti-RGNNV and anti-BFNNV sera reacted with both RGNNV and BFNNV strains. The present serological findings will be important in investigating the infectivity and host specificity of betanodaviruses and in developing vaccines for the disease. PMID- 14735918 TI - Infection and propagation of lymphocystis virus isolated from the cultured flounder Paralichthys olivaceus in grass carp cell lines. AB - The causative agent of lymphocystis disease that frequently occurs in cultured flounder Paralichthys olivaceus in China is lymphocystis virus (LV). In this study, 13 fish cell lines were tested for their susceptibility to LV. Of these, 2 cell lines derived from the freshwater grass carp Ctenopharyngodon idellus proved susceptible to the LV, and 1 cell line, GCO (grass carp ovary), was therefore used to replicate and propagate the virus. An obvious cytopathic effect (CPE) was first observed in cell monolayers at 1 d post-inoculation, and at 3 d this had extended to about 75% of the cell monolayer. However, no further CPE extension was observed after 4 d. Cytopathic characteristics induced by the LV were detected by Giemsa staining and fluorescence microscopic observation with Hoechst 33258 staining. The propagated virus particles were also observed by electron microscopy. Ultrastructure analysis revealed several distinct cellular changes, such as chromatin compaction and margination, vesicle formation, cell-surface convolution, nuclear fragmentation and the occurrence of characteristic 'blebs' and cell fusion. This study provides a detailed report of LV infection and propagation in a freshwater fish cell line, and presents direct electron microscopy evidence for propagation of the virus in infected cells. A possible process by which the CPEs are controlled is suggested. PMID- 14735919 TI - Acute and persistent experimental nodavirus infection in spotted wolffish Anarhichas minor. AB - Spotted wolffish Anarhichas minor (approx. 0.7 g) were found to be susceptible to infection with a nodavirus isolated from Atlantic halibut (AHNV) by bath challenge. During the acute stage of infection, 4 to 8 wk post-challenge, viral encephalopathy and retinopathy (VER) were diagnosed by histopathology, immunohistochemistry (IHC) and reverse transcriptase-polymerase chain reaction (RT-PCR). Accumulated mortality was 52% in the challenged group. The surviving fish were sampled 16 wk post-challenge, by which time they had grown to approximately 17 g. No clinical signs of VER were observed in these fish. RT-PCR examination revealed the presence of nodavirus in several organs of the survivors, but no immunopositive cells were detected by IHC. Nodavirus was reisolated from fish at the last sampling in SSN-1 cells, showing that nodavirus retains virulence in persistently infected wolffish for at least 16 wk post-bath challenge. PMID- 14735920 TI - Isolation of a rhabdovirus during outbreaks of disease in cyprinid fish species at fishery sites in England. AB - A virus was isolated during disease outbreaks in bream Abramis brama, tench Tinca tinca, roach Rutilis rutilis and crucian carp Carassius carassius populations at 6 fishery sites in England in 1999. Mortalities at the sites were primarily among recently introduced fish and the predominant fish species affected was bream. The bream stocked at 5 of the 6 English fishery sites were found to have originated from the River Bann, Northern Ireland. Most fish presented few consistent external signs of disease but some exhibited clinical signs similar to those of spring viraemia of carp (SVC), with extensive skin haemorrhages, ulceration on the flanks and internal signs including ascites and petechial haemorrhages. The most prominent histopathological changes were hepatocellular necrosis, interstitial nephritis and splenitis. The virus induced a cytopathic effect in tissue cultures (Epithelioma papulosum cyprini [EPC] cells) at 20 degrees C and produced moderate signals in an enzyme immunoassay (EIA) for the detection of SVC virus. The virus showed a close serological relationship to pike fry rhabdovirus in both EIA and serum neutralisation assays and to a rhabdovirus isolated during a disease outbreak in a bream population in the River Bann in 1998. A high degree of sequence similarity (> or = 99.5% nucleotide identity) was observed between the English isolates and those from the River Bann. Experimental infection of juvenile bream, tench and carp with EPC cell-grown rhabdovirus by bath and intraperitoneal injection resulted in a 40% mortality of bream in the injection group only. The virus was re-isolated from pooled kidney, liver and spleen tissue samples from moribund bream. The field observations together with the experimental results indicate that this rhabdovirus is of low virulence but may have the potential to cause significant mortality in fishes under stress. PMID- 14735921 TI - Construction of a safe, stable, efficacious vaccine against Photobacterium damselae ssp. piscicida. AB - Vaccination with bacterial auxotrophs, particularly those with an interruption in the common pathway of aromatic amino-acid biosynthesis, known as the shikimate pathway, has been shown to be effective in the prevention of a variety of bacterial diseases. In order to evaluate this approach to vaccine development in the important marine pathogen Photobacterium damselae subsp. piscicida, the aroA gene of the shikimate pathway was identified from a P. damselae subsp. piscicida genomic library by complementation in an aroA mutant of Escherichia coli. The complementing plasmid was isolated and the nucleotide sequence of the P. damselae subsp. piscicida genomic insert was determined. Subsequent analysis of the DNA sequence data demonstrated that the identified plasmid contained 3464 bp of P. damselae subsp. piscicida DNA, including the complete aroA gene. The sequence data was used to delete a 144 bp MscI fragment, and the kanamycin resistance gene (kan) from transposon Tn903 was ligated into the MscI site. This delta(aro)A::kan construct was sub-cloned into a suicide plasmid and transferred to a wild-type P. damselae subsp. piscicida by conjugation and allelic exchange. One selected mutant, LSU-P2, was confirmed phenotypically to require supplementation with aromatic metabolites for growth in minimal media, and was confirmed genotypically by PCR and DNA sequencing. Further, LSU-P2 was demonstrated to be avirulent in hybrid striped bass and to provide significant protection against disease following challenge with the wild-type strain. PMID- 14735922 TI - Protozoal and epitheliocystis-like infections in the introduced bluestripe snapper Lutjanus kasmira in Hawaii. AB - The bluestripe snapper, or taape, was introduced into Hawaii in the 1950s and has since become very abundant throughout the archipelago. As part of a health survey of reef fish in Hawaii, we necropsied 120 taape collected from various coastal areas south of Oahu and examined fish histology for extraintestinal organisms. Forty-seven percent of taape were infected with an apicomplexan protozoan compatible with a coccidian. Infection was evident mainly in the spleen and, less commonly, the kidney. Prevalence of this coccidian increased with size of fish, and we saw no significant pathology associated with the organism. Twenty-six percent of taape were also infected with an epitheliocystis-like organism that occurred mainly in the kidney and, less commonly, the spleen. In contrast to the coccidian, fish mounted a notable inflammatory response to the epitheliocystis like organism, and this inflammation appeared to increase in severity with age. Prevalence of the epitheliocystis-like organism infection increased with age, but infection was not seen in fish greater than 26.5 cm fork length. The high prevalence of coccidial infection in introduced taape prompts the concern that these organisms, along with the epitheliocystis-like organism, have the potential to be transmitted to native reef fish. Given the impact of other introduced microbial organisms on native Hawaiian fauna, there is a clear need to assess whether protozoa and bacteria are endemic to Hawaii, and whether they negatively impact native reef fish that closely associate with taape. PMID- 14735923 TI - Minor effect of depletion of resident macrophages from peritoneal cavity on resistance of common carp Cyprinus carpio to blood flagellates. AB - Carp Cyprinus carpio macrophages were depleted by intraperitoneal (i.p.) injection of clodronate-liposomes for the in vivo study of the effect of macrophage depletion on the resistance of carp to infection with blood flagellate parasites. Clodronate released inside the cell induces apoptosis of (murine) macrophages. Following i.p. injection of carp with liposomes alone, but not with Trypanoplasma borreli, neutrophilic granulocytes rapidly migrated from the head kidney to the peritoneal cavity. The majority of liposomes in the peritoneal cavity were not taken up by newly arrived neutrophilic granulocytes, however, but by resident macrophages. After 2 i.p. injections of clodronate-liposomes, the percentage of macrophages present in the peritoneal cavity was significantly reduced, as evaluated by flow cytometry. Macrophage-depleted carp that were infected i.p. with T. borreli suffered from high mortality. However, these fish did not show lethal parasitaemia but did show clear bacteraemia. Macrophage depleted carp that were infected i.p. with Trypanosoma carassii showed a minor increase in parasitaemia. In addition, macrophage-depleted carp, immune to T. borreli as a result of having survived a prior infection, remained immune to i.p. reinfection with T. borreli. Succesful depletion of peritoneal macrophages seemed to have a minor effect on the resistance of carp against blood flagellates. However, carp macrophages are essential as a first line of defence against (bacterial) infection. PMID- 14735924 TI - Efficacy of passive sand filtration in reducing exposure of salmonids to the actinospore of Myxobolus cerebralis. AB - The aquatic oligochaete Tubifex tubifex parasitized by Myxobolus cerebralis releases triactinomyxon (TAM) actinospores that can infect some species of salmonids and cause salmonid whirling disease. Silica sand was tested as a filtration medium for removal of TAMs from water containing the parasite. Laboratory tests indicated sand filtration removed > 99.99% of TAMs. In 2 different field tests, groups of 1 mo old rainbow trout Oncorhynchus mykiss were exposed for 2 wk to filtered and unfiltered water from a spring-fed pond enzootic for M. cerebralis. In November 2000, the exposure dose was estimated as between 3 and 5 TAMs fish(-1). During a March 2001 exposure, the estimated dose was between 286 and 404 TAMs fish(-1). Fish were held for 6 mo post exposure (p.e.) in laboratory aquaria for observation and evidence of clinical signs of whirling disease. We used 4 diagnostic techniques to assess the prevalence and severity of infection by M. cerebralis among fish exposed to filtered and unfiltered water. These included polymerase chain reaction (PCR) for genomic DNA of the parasite, histological evaluation for tissue damage, tissue digestion for quantification of cranial myxospores of the parasite, and total non-sampling mortality that occurred over 6 mo p.e. All diagnostic tests verified that the prevalence and severity of infection was significantly reduced among fish in treatment groups exposed to filtered water compared to those exposed to unfiltered water in both the low-dose and high-dose exposures. PMID- 14735925 TI - Identifying epidemiological factors affecting sea lice Lepeophtheirus salmonis abundance on Scottish salmon farms using general linear models. AB - The variation in Lepeophtheirus salmonis sea lice numbers across 40 Scottish salmon farm sites during 1996 to 2000 was analysed using mean mobile abundance for 3 important 6 mo periods within the production cycle. Using statistical regression techniques, over 20 management and environmental variables suspected to have an effect on controlling lice populations were investigated as potential risk factors. The findings and models developed provide a picture of mobile L. salmonis infestation patterns on Scottish farm sites collectively. The results identified level of treatment, type of treatment, cage volume, current speed, loch flushing time and sea lice levels in the preceding 6 mo period to be key explanatory factors. Factors such as stocking density, site biomass, water temperature and the presence of neighbours, previously cited to be important correlates of sea lice risk from analysis of individual sites over time, were not found to be important. Variation in mobile abundance in the first half of the second year of production could be adequately explained (adjusted R2 between 55 and 72%) by the recorded data, suggesting that there is scope for management to control L. salmonis abundance, though much of the variation remains unexplained. PMID- 14735926 TI - Life cycle of Ceratothoa oestroides, a cymothoid isopod parasite from sea bass Dicentrarchus labrax and sea bream Sparus aurata. AB - Ceratothoa oestroides (Risso, 1826) (Isopoda: Cymothoida) is a protandric hermaphrodite parasite on a wide range of wild fish species. In recent years it has become a threat to cage-reared fish facilities, where high fish density provides optimal conditions for transmission. Its impact on fish health and economical gain is significant, varying from growth retardation and decreased immunocompetency to direct loss due to mass mortalities of juvenile fishes. Because of the sheltered location of the parasite in the buccal cavity of fishes, chemotherapeutics are ineffective. An understanding of the C. oestroides life cycle and its behavioral mechanisms could prove constructive tools for the prevention and control of infection. This study describes the reproductive cycle of C. oestroides experimentally induced in different fish hosts and temperature regimes. Sea bream larvae Sparus aurata and 1 yr annular sea bream Diplodus annularis were chosen as experimental models, and were held at 22 and 19.5 degrees C, respectively. The reproductive cycle of S. aurata was not completed within 4 mo (at which point the last larva died of severe anemia and respiratory distress), while that of the annular sea bream was completed successfully after 1 mo. PMID- 14735927 TI - Heart morphology in wild and farmed Atlantic salmon Salmo salar and rainbow trout Oncorhynchus mykiss. AB - The normal shape of the salmonid ventricle is a triangular pyramid with the apex pointing caudoventrally. A strong positive correlation has been established between this shape and optimum cardiac output and function. Domesticated salmonids appear to have developed a more rounded ventricle with misaligned bulbus arteriosus. Several reports from fish health veterinarians indicate that fish with abnormal heart morphology have a high mortality rate during stress inducing situations like grading, transportation and bath treatments. The present paper compares and describes the ventricle morphology of wild vs. farmed Atlantic salmon, and wild steelhead (anadromous rainbow trout) vs. farmed rainbow trout. Several parameters were measured to provide numerical measurement of the differences in shape, i.e. height:width ratio and the angle between the longitudinal ventricular axis and the axis of the bulbus arteriosus. We conclude that the hearts of farmed fish are rounder than those in corresponding wild fish, and that the angle between the ventricular axis and the axis of the bulbus arteriosus is more acute in wild fish than in their farmed counterparts. Further studies are necessary to reveal the prevalence, functional significance and possible causes of these abnormal hearts. PMID- 14735928 TI - Reduction of adhesion properties of Ruditapes philippinarum hemocytes exposed to Vibrio tapetis. AB - Vibrio tapetis is the causative agent of brown ring disease (BRD), which affects a species of clam, Ruditapes philippinarum. After incubation with V. tapetis, hemocytes lose filopods and become rounded, indicating cytotoxic activity of the bacterium. To rapidly quantify this cytotoxicity, a flow-cytometry test was developed based on the capacity of V. tapetis to inhibit adhesion of clam hemocytes to plastic. Several bacteria:hemocyte ratios, the cytotoxicity of other Vibrio spp. pathogenic to bivalves, and that of various V. tapetis isolates were tested. Inhibition of adherence is detectable with as few as 5 bacteria per hemocyte. The greater cytotoxic activity of V. tapetis compared to that of V. splendidus and V. pectenicida suggests a specific pathogenicity of V. tapetis to R. philippinarum hemocytes. Although all V. tapetis isolates inhibited adhesion, significant variations in cytotoxicity among isolates was demonstrated. PMID- 14735929 TI - Isolation and characterization of serine protease gene(s) from Perkinsus marinus. AB - This study reports the first serine protease gene(s) isolated from Perkinsus marinus. Using universal primers, a 518 bp subtilisin-like serine protease gene fragment was amplified from P. marinus genomic DNA and used as a probe to screen a lambda-phage P. marinus genomic library; 2 different lambda-phage clones hybridized to the digoxigenin(DIG)-labeled subtilisin-like gene fragment. Following subcloning and sequencing of the larger DNA fragment, a 1254 bp open reading frame was identified and later confirmed, by 5' and 3' random amplification of cDNA ends (RACE) and northern blot analysis, to contain the entire coding-region sequence. Sequence analysis of the 3' RACE results from 2 isolate cultures, VA-2 (P-1) and LA 10-1, revealed multiple polymorphic sites within and among isolates. We identified 2 different types of cDNA clones with 95.53% nucleotide sequence similarity, suggesting the possibility of 2 closely related genes within the P. marinus genome. Southern blot analysis of genomic DNA from 12 genetically distinct P. marinus isolate cultures revealed 2 different banding patterns among isolates. PMID- 14735930 TI - Effects of shell lesions on survival, growth, condition and reproduction in the New Zealand blackfoot abalone Haliotis iris. AB - The pathogenicity of shell lesions in Haliotis iris Martyn was examined in a laboratory experiment in which 73 apparently healthy and 106 lesion-bearing abalone were maintained for up to 12 mo. The abalone were collected from the wild and kept in cages (1 ind. cage(-1)) for 4, 8 or 12 mo, at which times estimates of survival, growth, condition and reproductive capacity were made for each of 3 groups: 'healthy' (n = 73), 'mildly affected' (n = 61) and 'severely affected' (n = 32). Unaffected abalone showed a 2.7% mortality (n = 73) compared to 7.5% (n = 93) in lesion-bearing individuals over the entire experiment. Growth rates were significantly decreased in mildly and severely affected abalone: the relative von Bertalanffy growth coefficient (K), calculated over 12 mo, was -0.176 for unaffected, -0.079 for mildly affected and -0.048 for severely affected individuals. The asymptotic length (L(infinity)) was calculated to be 131.5 mm for unaffected, 142.1 mm for mildly affected and 150.3 mm for severely affected abalone. Significantly (p < 0.05) lower condition indices and decreased reproductive capacity (p > 0.05) were obtained for the severely affected group compared to unaffected abalone. These trends were consistent over the course of the experiment. PMID- 14735932 TI - Detection by PCR of hepatopancreatic parvovirus (HPV) and other viruses in hatchery-reared Penaeus monodon postlarvae. AB - The prevalence of hepatopancreatic parvovirus (HPV), monodon baculovirus (MBV) and white spot syndrome virus (WSSV) in samples of Penaeus monodon postlarvae (PL10 to PL20, 10 to 20 d old postlarvae) in India was studied by PCR. Samples collected from different hatcheries, and also samples submitted by farmers from different coastal states, were analyzed. HPV was detected in 34%) of the hatchery samples and 31% of the samples submitted by farmers, using a primer set designed for detection of HPV from P. monodon in Thailand. However, none of these samples were positive using primers designed for detection of HPV from P. chinensis in Korea. This indicated that HPV from India was more closely related to HPV from P. monodon in Thailand. MBV was detected in 64% of the samples submitted by the farmers and 71% of the hatchery samples. A total of 84 % of the samples submitted by farmers, and 91% of the hatchery samples, were found positive for WSSV. Prevalence of concurrent infections by HPV, MBV and WSSV was 27% in hatchery samples and 29%, in samples submitted by farmers. Only 8% of the hatchery samples and 16% of the samples submitted by farmers were negative for all 3 viruses. This is the first report on the prevalence of HPV in P. monodon postlarvae from India. PMID- 14735931 TI - Infection of the cockle Cerastoderma edule in the Baie des Veys (France) by the microsporidian parasite Steinhausia sp. AB - We report the occurrence of the microsporidian parasite Steinhausia sp. in the oocytes of the common cockle Cerastoderma edule in a natural population in France, where high mortalities occurred. Steinhausia sp. appeared primarily as sporocysts containing many small spores, and putative earlier developmental stages were also observed. Both its prevalence and infection intensity were low, and no host defence reaction was recognized, suggesting that Steinhausia sp. had no detrimental effect on C. edule. Its prevalence was higher in cockles lying on the sediment surface, but the significance of this observation could not be explained given the poor knowledge of the Steinhausia life cycle. The present data did not allow specific identification of the parasite, and further studies are required to determine whether Steinhausia sp. in the cockle is a new species, or a microsporidian infecting multiple host species. PMID- 14735933 TI - Vibrio alginolyticus associated with white spot disease of Penaeus monodon. AB - In February 2000, white spot disease outbreaks occurred among cultured Penaeus monodon in extensive shrimp farms on the southwest coast of India. Bacteria were isolated from infected shrimp that showed reddish body coloration and white spots in the cuticle. The isolates were screened on thiosulfate citrate bile salt sucrose (TCBS) agar plates for the selection of Vibrio species. The primary isolate (QS7) was characterized as V. alginolyticus based on morphological, biochemical and physiological characteristics. Antibiotic sensitivity tests of QS7 indicated that the isolate was highly sensitive to chloramphenicol, ciprofloxacin, nalidixic acid and streptomycin. Pathogenicity tests confirmed that the isolate was virulent for P. monodon. Based on the lethal dose (LD50) value (5 x 10(6) cfu per shrimp), it was inferred that shrimp weakened by white spot syndrome virus would succumb to secondary infection by QS7. PMID- 14735934 TI - Effect of potassium permanganate stress on immune resistance and susceptibility to Lactococcus garvieae in the giant freshwater prawn Macrobrachium rosenbergii. AB - This work is part of a continuing series of investigations on the effect of commonly used aquaculture chemicals on the immune resistance and susceptibility of the giant freshwater prawn Macrobrachium rosenbergii to Lactococcus garvieae. The methodology has been described in earlier publications of the series. Potassium permanganate at 1.0 mg l(-1) in tryptic soy broth (TSB) had no effect on the growth rate of L. garvieae. The mortality of M. rosenbergii challenged with 4 x 10(6) colony-forming units (cfu) prawn(-1) of TSB-grown L. garvieae was significantly greater than that of challenged controls. Addition of potassium permanganate at 1.0 mg l(-1) in TSB significantly increased the virulence of L. garvieae to M. rosenbergii. Exposure of M. rosenbergii to potassium permanganate prior to challenge with TSB-grown L. garvieae at 4 x 10(6) and 3 x 10(6) cfu prawn(-1) revealed that 96 h mortality was significantly lower for prawns held in water containing 0.3 mg l(-1) of the chemical than for prawns in water containing 1.0 mg l(-1) or no chemical. Potassium permanganate caused no significant changes in total hemocyte counts and differential hemocyte counts, compared to the control treatments. However, a concentration of 1.0 mg l(-1) or more for 96 h resulted in decreased phenoloxidase activity, phagocytic activity and clearance efficiency. Respiratory burst increased with exposure to 0.3 mg l(-1). In conclusion, treatment with potassium permanganate at 0.3 mg 1(-1) was effective in reducing M. rosenbergii mortality from L. garvieae infection, but higher concentrations had a negative effect, probably due to reduced prawn defenses. PMID- 14735935 TI - Experimental infection of twenty species of Indian marine crabs with white spot syndrome virus (WSSV). AB - Twenty species of Indian marine crabs were experimentally infected with white spot syndrome virus (WSSV), via the oral route and intramuscular injection, to determine their viral susceptibility. We determined that 16 species (Calappa philargius, Charybdis annulata, C. lucifera, Doclea hybrida, Grapsus albolineatus, Halimede ochtodes, Liagore rubronaculata, Lithodes maja, Matuta miersi, Paradorippe granulata, Parthenope prensor, Philyra syndactyla, Podophthalmus vigil, Portunus sanquinolentus, Scylla serrata and Thalamita danae) were susceptible and 4 (Atergatis integerrimus, Charybdis natator, Demania splendida or Menippe rumphii) were refractive at 50 d post-infection (p.i.). The presence of WSSV in these crabs was confirmed by PCR tests, histology and bioassay. WSSV was found in the gill, heart, eyestalks, striated muscle and cephalothoraxic tissue. The 4 WSSV-refractive species represent potential reservoirs or carriers of WSSV. PMID- 14735936 TI - Effect of epidermal papillomatosis on survival of the freshwater fish Rutilus rutilus. AB - Epidermal papillomatosis occurs in several marine and freshwater fish species. Previously, papillomatosis has been shown to induce mortality in juvenile carp. We studied the effect of epidermal papillomatosis on the survival of adult male roach Rutilus rutilus by caging naturally diseased, marked (by us) fish in the field. Within the constraints of the experimental design, there was no difference in survival between healthy, slightly diseased and heavily diseased fish. Therefore, we conclude that the possible effect of epidermal papillomatosis on the mortality of wild roach is relatively minor. PMID- 14735937 TI - Association of Type III secretion genes with virulence of Aeromonas salmonicida subsp. salmonicida. AB - Aeromonas salmonicida subsp. salmonicida possesses a number of potential virulence factors, including a recently identified plasmid-encoded Type III secretion system. A number of field isolates of A. salmonicida subsp. salmonicida were examined for the presence of Type III secretion genes. Using in vitro experiments, it was found that field isolates containing such genes are cytotoxic to fish cell lines, whereas those that lack these genes are not. Using a rainbow trout in vivo model, the virulence of a wild type A. salmonicida subsp. salmonicida strain (Strain JF2267), which possesses Type III secretion genes, was compared to that of a laboratory derivative of the same strain that has lost these genes. While Strain JF2267 was virulent towards rainbow trout, its derivative was not. The A. salmonicida subsp. salmonicida Type Strain ATCC 33658T, which also lacks Type III secretion genes, was also found to be avirulent by this challenge model. The findings from both the in vitro and in vivo experiments suggest that the presence of Type III secretion genes is associated with the virulence of this important fish pathogen. PMID- 14735938 TI - First record of Huffmanela schouteni (Nematoda: Trichosomoididae), a histozoic parasite of flyingfishes, in Europe. AB - Examination of 3 specimens of the Mediterranean flyingfish Cheilopogon heterurus (Rafinesque) from the Ligurian Sea, Italy, revealed the presence of numerous, dark-shelled trichinelloid eggs (69 to 75 x 30 to 33 microm) inside the innermost layer of the swimbladder, mostly located in clumps arranged in a distinct pattern. These were identified as Huffmanela schouteni Moravec et Campbell, 1991, a species known so far as eggs in flyingfishes and in the stools of humans in Curacao. This is the first finding of this parasite in Europe and C. heterurus represents a new host record. PMID- 14735939 TI - Lernanthropus kroyeri infections in farmed sea bass Dicentrarchus labrax: pathological features. AB - Twenty sea bass Dicentrarchus labrax L. from a fish farm (floating cage) in Greece were examined for the presence of parasites. The gills of 7 (35%) fish were infected with adult female specimens of the parasitic copepod Lernanthropus kroyeri van Beneden, 1851, and the intensity of infection ranged from 1 to 24 parasites per host. The most infected portion of the gills appeared to be the primary lamellae. Erosion, desquamation and necrosis of the secondary lamellae were noticed near the site of copepod attachment; furthermore, the terminal claw of the second antennae lacerated tissue and vessels of infected gill. Parasitism by L. kroyeri affected the host's condition factor (mean +/- SE in uninfected vs parasitized; 1.88 +/- 0.04 vs 1.66 +/- 0.12; p < 0.05). PMID- 14735940 TI - Using texture to analyze and manage large collections of remote sensed image and video data. AB - We describe recent research into using the visual primitive of texture to analyze and manage large collections of remote sensed image and video data. Texture is regarded as the spatial dependence of pixel intensity. It is characterized by the amount of dependence at different scales and orientations, as measured with frequency-selective filters. A homogeneous texture descriptor based on the filter outputs is shown to enable (1) content-based image retrieval in large collections of satellite imagery, (2) semantic labeling and layout retrieval in an aerial video management system, and (3) statistical object modeling in geographic digital libraries. PMID- 14735941 TI - Building extraction from stereoscopic aerial images. AB - Three-dimensional models of urban objects are widely used in geographic information systems, telecommunications, or defense applications. The classic technique for obtaining such models is stereoscopy. Images are densely matched, and images of above-ground structures are delineated. We propose two semiautomatic methods based on the Hough transform and statistically active models to delineate buildings. The first one delineates rectangular shapes; the second one deals with more-complex buildings. Each one is based on a criterion optimization that takes both photometric and altimetric information into account. Results based on real data show that the first method is robust and that the second one, which deals with a broad range of buildings, seems to be a good compromise between robustness and applicability. PMID- 14735942 TI - Feature reduction and morphological processing for hyperspectral image data. AB - An automatic target detection system that uses hyperspectral (HS) imagery is proposed. HS images contain both spatial and spectral response information that provides detailed descriptions of an object. These new, to our knowledge, sensor data are useful in automatic target recognition applications. To provide discrimination information from the HS images and to select features that generalize well, we describe a new, to our knowledge, high-dimensional generalized discriminant feature-extraction algorithm and compare its performance with that of other feature-reduction methods for two HS target detection applications (mine and vehicle detection) by using a nearest-neighbor classifier. We also advance an approach to simultaneously optimize both spatial and spectral responses. PMID- 14735943 TI - Model-based approach to the detection and classification of mines in sidescan sonar. AB - This paper presents a model-based approach to mine detection and classification by use of sidescan sonar. Advances in autonomous underwater vehicle technology have increased the interest in automatic target recognition systems in an effort to automate a process that is currently carried out by a human operator. Current automated systems generally require training and thus produce poor results when the test data set is different from the training set. This has led to research into unsupervised systems, which are able to cope with the large variability in conditions and terrains seen in sidescan imagery. The system presented in this paper first detects possible minelike objects using a Markov random field model, which operates well on noisy images, such as sidescan, and allows a priori information to be included through the use of priors. The highlight and shadow regions of the object are then extracted with a cooperating statistical snake, which assumes these regions are statistically separate from the background. Finally, a classification decision is made using Dempster-Shafer theory, where the extracted features are compared with synthetic realizations generated with a sidescan sonar simulator model. Results for the entire process are shown on real sidescan sonar data. Similarities between the sidescan sonar and synthetic aperture radar (SAR) imaging processes ensure that the approach outlined here could be made applied to SAR image analysis. PMID- 14735944 TI - From snakes to region-based active contours defined by region-dependent parameters. AB - Image and sequence segmentation of a the segmentation task are discussed from the point of view of optimizing the segmentation criterion. Such a segmentation criterion involves so-called (boundary and region) descriptors, which, in general, may depend on their respective boundaries or regions. This dependency must be taken into account when one is computing the criterion derivative with respect to the unknown object domain (defined by its boundary). If this dependency not considered, some correctional terms may be omitted. Computing the derivative of the segmentation criterion with a dynamic scheme is described. The scheme is general enough to provide a framework for a wide variety of applications in segmentation. It also provides a theoretical meaning to the philosophy of active contours. PMID- 14735945 TI - Point target detection and subpixel position estimation in optical imagery. AB - We address the issue of distinguishing point objects from a cluttered background and estimating their position by image processing. We are interested in the specific context in which the object's signature varies significantly relative to its random subpixel location because of aliasing. The conventional matched filter neglects this phenomenon and causes a consistent degradation of detection performance. Thus alternative detectors are proposed, and numerical results show the improvement brought by approximate and generalized likelihood-ratio tests compared with pixel-matched filtering. We also study the performance of two types of subpixel position estimator. Finally, we put forward the major influence of sensor design on both estimation and point object detection. PMID- 14735946 TI - Detection of linear features in synthetic-aperture radar images by use of the localized Radon transform and prior information. AB - A new linear-features detection method is proposed for extracting straight edges and lines in synthetic-aperture radar images. This method is based on the localized Radon transform, which produces geometrical integrals along straight lines. In the transformed domain, linear features have a specific signature: They appear as strongly contrasted structures, which are easier to extract with the conventional ratio edge detector. The proposed method is dedicated to applications such as geographical map updating for which prior information (approximate length and orientation of features) is available. Experimental results show the method's robustness with respect to poor radiometric contrast and hidden parts and its complementarity to conventional pixel-by-pixel approaches. PMID- 14735947 TI - Target detection with a liquid-crystal-based passive Stokes polarimeter. AB - We present an imaging system that measures the polarimetric state of the light coming from each point of a scene. This system, which determines the four components of the Stokes vector at each spatial location, is based on a liquid crystal polarization modulator, which makes it possible to acquire four dimensional Stokes parameter images at a standard video rate. We show that using such polarimetric images instead of simple intensity images can improve target detection and segmentation performance. PMID- 14735948 TI - Clustering of polarization-encoded images. AB - Polarization-encoded imaging consists of the distributed measurements of polarization parameters for each pixel of an image. We address clustering of multidimensional polarization-encoded images. The spatial coherence of polarization information is considered. Two methods of analysis are proposed: polarization contrast enhancement and a more-sophisticated image-processing algorithm based on a Markovian model. The proposed algorithms are applied and validated with two different Mueller images acquired by a fully polarimetric imaging system. PMID- 14735949 TI - Building a cascade detector and its applications in automatic target detection. AB - A hierarchical classifier (cascade) is proposed for target detection. In building an optimal cascade we considered three heuristics: (1) use of a frontier following approximation, (2) controlling error rates, and (3) weighting. Simulations of synthetic data with various underlying distributions were carried out. We found that a weighting heuristic is optimal in terms of both computational complexity and error rates. We initiate a systematic comparison of several potential heuristics that can be utilized in building a hierarchical model. A range of discussions regarding the implications and the promises of cascade architecture as well as of techniques that can be integrated into this framework is provided. The optimum heuristic--weighting algorithms--was applied to an IR data set. It was found that these algorithms outperform some state-of the-art approaches that utilize the same type of simple classifier. PMID- 14735950 TI - Constrained quadratic correlation filters for target detection. AB - A method for designing and implementing quadratic correlation filters (QCFs) for shift-invariant target detection in imagery is presented. The QCFs are quadratic classifiers that operate directly on the image data without feature extraction or segmentation. In this sense the QCFs retain the main advantages of conventional linear correlation filters while offering significant improvements in other respects. Not only is more processing required for detection of peaks in the outputs of multiple linear filters but choosing the most suitable among them is an error-prone task. All channels in a QCF work together to optimize the same performance metric and to produce a combined output that leads to considerable simplification of the postprocessing scheme. The QCFs that are developed involve hard constraints on the output of the filter. Inasmuch as this design methodology is indicative of the synthetic discriminant function (SDF) approach for linear filters, the filters that we develop here are referred to as quadratic SDFs (QSDFs). Two methods for designing QSDFs are presented, an efficient architecture for achieving them is discussed, and results from the Moving and Stationary Target Acquisition and Recognition synthetic aperture radar data set are presented. PMID- 14735951 TI - Infrared target detection with probability density functions of wavelet transform subbands. AB - We report the development of a wavelet multiresolution texture-based algorithm that uses the probability density functions (PDFs) of the subband of the wavelet decomposition of an image. The moments of these pdfs are used in a clustering algorithm to segment the targets from their background clutter. Using the tools of experimental methodology, we evaluate the performance of this algorithm on real infrared imagery under varying algorithm parameter sets as well as scene, image, and false-alarm conditions. We estimate a set of multidimensional predictive analytic performance models that relate the detection probabilities as functions of false alarm, algorithm internal parameter, target pixel number, target-to-background interference ratio, target-interference ratio, and Fechner Weber and local entropy metrics in the scene. These models can be used to predict performance in regions were no data are available and to optimize performance by selection of the optimum parameter and constant false-alarm values in regions with known scene and metric conditions. PMID- 14735952 TI - Detecting three-dimensional location and shape of noisy distorted three dimensional objects with ladar trained optimum nonlinear filters. AB - We propose a filtering technique that uses laser radar (ladar) data to detect a target's three-dimensional (3D) coordinates and shape within an input scene. A two-dimensional ladar range image is converted into 3D space, and then the 3D optimum nonlinear filtering technique is used to detect the 3D coordinates of targets (including the target's distance from the sensor). The 3D optimum nonlinear filter is designed to detect distorted targets (i.e., out-of-plane and in-plane rotations and scale changes) and to be noise robust. The nonlinear filter is derived to minimize the mean of the output energy in response to the input scene in the presence of disjoint background noise and additive noise and to maintain a fixed output peak for the members of the true-class target training set. The system is tested with real ladar imagery in the presence of background clutter. The background clutter used in the system evaluation includes false objects that are similar to the true targets. The correlation output of ladar images shows a dominant peak at the target's 3D coordinates. PMID- 14735953 TI - Automated vehicle detection in forward-looking infrared imagery. AB - We describe an algorithm for the detection and clutter rejection of military vehicles in forward-looking infrared (FLIR) imagery. The detection algorithm is designed to be a prescreener that selects regions for further analysis and uses a spatial anomaly approach that looks for target-sized regions of the image that differ in texture, brightness, edge strength, or other spatial characteristics. The features are linearly combined to form a confidence image that is thresholded to find likely target locations. The clutter rejection portion uses target specific information extracted from training samples to reduce the false alarms of the detector. The outputs of the clutter rejecter and detector are combined by a higher-level evidence integrator to improve performance over simple concatenation of the detector and clutter rejecter. The algorithm has been applied to a large number of FLIR imagery sets, and some of these results are presented here. PMID- 14735954 TI - Real-time target recognition and tracking of vehicles moving on a street by matched filtering that uses quantization of modulated function to complexes of a quadrupole. AB - Vehicles moving on a street were monitored by a CCD digital TV camera and recognized in real time by a nonlinear matched filtering method, quantization of modulated function to complexes of a quadrupole, with a personal computer. In the nonlinear matched filtering method an input scene containing the vehicles is modulated in the frequency domain by matched filters, the modulated functions are quantized to complex numbers representing a quadrupole, and then quantized functions are inverse-Fourier transformed. In the frame time of an NTSC-TV system, autocorrelation signals of four vehicles were calculated and discriminated, and their peak positions were determined. Their correct loci in real time were tracked in sequential frames. PMID- 14735955 TI - Heteroassociative multiple-target tracking by fringe-adjusted joint transform correlation. AB - A heteroassociative joint transform correlation (JTC) technique is proposed for recognizing and tracking multiple heteroassociative or dissimilar targets from gray-level image sequences by use of the concept of fringe-adjusted JTC and a multiple-target-detection algorithm. A fringe-adjusted JTC technique is used to ensure quantification of the similarities among several input images while it satisfies the equal-correlation-peak criterion. Tracking is accomplished by retrieval of the target motion information estimated from multiple consecutive image frames. An enhanced version of the fringe-adjusted filter is incorporated into the heteroassociative multiple-target-detection process to optimize the correlation performance. The feasibility of the proposed technique is tested by computer simulation with real infrared image data. PMID- 14735956 TI - Three-dimensional tracking of multiple skin-colored regions by a moving stereoscopic system. AB - A system that performs three-dimensional (3D) tracking of multiple skin-colored regions (SCRs) in images acquired by a calibrated, possibly moving stereoscopic rig is described. The system consists of a collection of techniques that permit the modeling and detection of SCRs, the determination of their temporal association in monocular image sequences, the establishment of their correspondence between stereo images, and the extraction of their 3D positions in a world-centered coordinate system. The development of these techniques has been motivated by the need for robust, near-real-time tracking performance. SCRs are detected by use of a Bayesian classifier that is trained with the aid of a novel technique. More specifically, the classifier is bootstrapped with a small set of training data. Then, as new images are being processed, an iterative training procedure is employed to refine the classifier. Furthermore, a technique is proposed to enable the classifier to cope with changes in illumination. Tracking of SCRs in time as well as matching of SCRs in the images of the employed stereo rig is performed through computationally inexpensive and robust techniques. One of the main characteristics of the skin-colored region tracker (SCRT) instrument is its ability to report the 3D positions of SCRs in a world-centered coordinate system by employing a possibly moving stereo rig with independently verging CCD cameras. The system operates on images of dimensions 640 x 480 pixels at a rate of 13 Hz on a conventional Pentium 4 processor at 1.8 GHz. Representative experimental results from the application of the SCRT to image sequences are also provided. PMID- 14735957 TI - Virtual three-dimensional blackboard: three-dimensional finger tracking with a single camera. AB - We present a method for three-dimensional (3D) tracking of a human finger from a monocular sequence of images. To recover the third dimension from the two dimensional images, we use the fact that the motion of the human arm is highly constrained owing to the dependencies between elbow and forearm and the physical constraints on joint angles. We use these anthropometric constraints to derive a 3D trajectory of a gesticulating arm. The system is fully automated and does not require human intervention. The system presented can be used as a visualization tool, as a user-input interface, or as part of some gesture-analysis system in which 3D information is important. PMID- 14735958 TI - Biometric verification with correlation filters. AB - Using biometrics for subject verification can significantly improve security over that of approaches based on passwords and personal identification numbers, both of which people tend to lose or forget. In biometric verification the system tries to match an input biometric (such as a fingerprint, face image, or iris image) to a stored biometric template. Thus correlation filter techniques are attractive candidates for the matching precision needed in biometric verification. In particular, advanced correlation filters, such as synthetic discriminant function filters, can offer very good matching performance in the presence of variability in these biometric images (e.g., facial expressions, illumination changes, etc.). We investigate the performance of advanced correlation filters for face, fingerprint, and iris biometric verification. PMID- 14735959 TI - Target detection and recognition improvements by use of spatiotemporal fusion. AB - We developed spatiotemporal fusion techniques for improving target detection and automatic target recognition. We also investigated real IR (infrared) sensor clutter noise. The sensor noise was collected by an IR (256 x 256) sensor looking at various scenes (trees, grass, roads, buildings, etc.). More than 95% of the sensor pixels showed near-stationary sensor clutter noise that was uncorrelated between pixels as well as across time frames. However, in a few pixels (covering the grass near the road) the sensor noise showed nonstationary properties (with increasing or decreasing mean across time frames). The natural noise extracted from the IR sensor, as well as the computer-generated noise with Gaussian and Rayleigh distributions, was used to test and compare different spatiotemporal fusion strategies. Finally, we proposed two advanced detection schemes: the double-thresholding the reverse-thresholding techniques. These techniques may be applied to complicated clutter situations (e.g., very-high clutter or nonstationary clutter situations) where the traditional constant-false-alarm ratio technique may fail. PMID- 14735960 TI - Multistage classification and recognition that employs vector quantization coding and criteria extracted from nonorthogonal and preprocessed signal representations. AB - Classification decision tree algorithms have recently been used in pattern recognition problems. In this paper, we propose a self-designing system that uses the classification tree algorithms and that is capable of recognizing a large number of signals. Preprocessing techniques are used to make the recognition process more effective. A combination of the original, as well as the preprocessed, signals is projected into different transform domains. Enormous sets of criteria that characterize the signals can be developed from the signal representations in these domains. At each node of the classification tree, an appropriately selected criterion is optimized with respect to desirable performance features such as complexity and noise immunity. The criterion is then employed in conjunction with a vector quantizer to divide the signals presented at a particular node in that stage into two approximately equal groups. When the process is complete, each signal is represented by a unique composite binary word index, which corresponds to the signal path through the tree, from the input to one of the terminal nodes of the tree. Experimental results verify the excellent classification accuracy of this system. High performance is maintained for both noisy and corrupt data. PMID- 14735961 TI - Nonlinear pattern recognition correlators based on color-encoding single-channel systems. AB - In color pattern recognition, color channels are normally processed separately and afterward the correlation outputs are combined. This is the definition of multichannel processing. We combine a single-channel method with nonlinear filtering based on nonlinear correlations. These nonlinear correlations yield better discrimination than common matched filtering. The method codes color information as amplitude and phase distributions and is followed by correlations related to binary decompositions. The technique is based on binary decompositions of the red, green, and blue and the hue, saturation, and intensity monochromatic channels of the reference and of the input scene, after which the binary information on the red, green, and blue channels and that of the hue, saturation, and intensity channels are encoded as different angles of a phase distribution. We have applied the method to images degraded by high levels of substitutive noise. Results show that the sliced orthogonal nonlinear generalized correlation detects the target with a high degree of discrimination when other methods fail. PMID- 14735962 TI - Recognition of polychromatic three-dimensional objects. AB - We propose to use optical multichannel correlation in various chromatic systems to obtain a setup for recognition of polychromatic three-dimensional (3-D) objects based on Fourier-transform profilometry. Because red-green-blue color components are not able to split the luminance information of objects in a defined component, when the 3-D objects are brighter than the reference objects the correlation result gives false alarms. We demonstrate that it is possible to use different color spaces that can split luminance from chromatic information to yield adequate recognition of polychromatic 3-D objects. We show experimental results that prove the utility of the proposed method. PMID- 14735963 TI - Three-dimensional object feature extraction and classification with computational holographic imaging. AB - We address three-dimensional (3D) object classification with computational holographic imaging. A 3D object can be reconstructed at different planes by use of a single hologram. We apply principal component and Fisher linear discriminant analyses based on Gabor-wavelet feature vectors to classify 3D objects measured by digital interferometry. Experimental and simulation results are presented for regional filtering concentrated at specific positions and for overall grid filtering. The proposed technique substantially reduces the dimensionality of the 3D classification problem. To the best of our knowledge, this is the first report on the use of the proposed technique for 3D object classification. PMID- 14735964 TI - Comparison of passive ranging integral imaging and active imaging digital holography for three-dimensional object recognition. AB - We present an overview of three-dimensional (3D) object recognition techniques that use active sensing by interferometric imaging (digital holography) and passive sensing by integral imaging. We describe how each technique can be used to retrieve the depth information of a 3D scene and how this information can then be used for 3D object recognition. We explore various algorithms for 3D recognition such as nonlinear correlation and target distortion tolerance. We also provide a comparison of the advantages and disadvantages of the two techniques. PMID- 14735965 TI - Multichip module with planar-integrated free-space optical vector-matrix-type interconnects. AB - Even in the semiconductor industry, free-space optical technology is nowadays seen as a prime option for solving the continually aggravating problem with VLSI chips, namely, that the interconnect technology has failed to keep pace with the increase in communication volume. To make free-space optics compatible with established lithography-based design and fabrication techniques the concept of planar integration was proposed approximately a decade ago. Here its evolution into a photonic microsystems engineering concept is described. For demonstration, a multichip module with planar-integrated freespace optical vector-matrix-type interconnects was designed and built. It contains flip-chip-bonded vertical cavity surface emitting laser arrays and a hybrid chip with an array of multiple quantum-well p-i-n diodes on top of a standard complementary metal-oxide semiconductor circuit as key optoelectronic hardware components. The optical system is integrated into a handy fused-silica substrate and fabricated with surface-relief diffractive phase elements. It has been optimized for the given geometrical and technological constraints and provides a good interconnection performance, as was verified in computer simulations on the basis of ray tracing and in practical experiments. PMID- 14735966 TI - Atmospheric modulation transfer function in the infrared. AB - In high-resolution ultranarrow field-of-view thermal imagers, image quality over relatively long path lengths is typically limited by atmospheric degradation, especially atmospheric blur. We report our results and analyses of infrared images from two sites, Fort A. P. Hill and Aberdeen Proving Ground. The images are influenced by the various atmospheric phenomena: scattering, absorption, and turbulence. A series of experiments with high-resolution equipment in both the 3 5- and 8-13-microm regions at the two locations indicate that, as in the visible, image quality is limited much more by atmosphere than by the instrumentation for ranges even of the order of only a few kilometers. For paths close to the ground, turbulence is more dominant, whereas for paths involving higher average elevation, aerosol modulation transfer function (MTF) is dominant. As wavelength increases, turbulence MTF also increases, thus permitting aerosol MTF to become more dominant. A critical role in aerosol MTF in the thermal infrared is attributed to absorption, which noticeably decreases atmospheric transmission much more than in the visible, thereby reducing high-spatial-frequency aerosol MTF. These measurements indicate that atmospheric MTF should be a basic component in imaging system design and analysis even in the infrared, especially as higher resolution hardware becomes available. PMID- 14735967 TI - Spectral Fraunhofer regime: time-to-frequency conversion by the action of a single time lens on an optical pulse. AB - We analyze a new regime in the interaction between an optical pulse and a time lens (spectral Fraunhofer regime), where the input pulse amplitude is mapped from the time domain into the frequency domain (time-to-frequency conversion). Here we derive in detail the conditions for achieving time-to-frequency conversion with a single time lens (i.e., for entering the spectral Fraunhofer regime) as well as the expressions governing this operation. Our theoretical findings are demonstrated both numerically and experimentally. A comparative study between the proposed single-time-lens configuration and the conventional dispersion + time lens configuration for time-to-frequency conversion is also conducted. Time-to frequency conversion with a single time lens can be used for applications similar to those previously proposed for the conventional time-to-frequency converters, e.g., high-resolution measurement of fast optical temporal waveforms. Moreover, our results also indicate that the spectral Fraunhofer regime provides additional capabilities for controlling and processing optical pulses. PMID- 14735968 TI - Optical planar and channel waveguides in the new nonlinear crystal Ca4YO(BO3)3 (YCOB) fabricated by He+ implantation. AB - We report the first study of optical planar and channel waveguides fabricated in the new nonlinear crystal Ca4YO(BO3) by use of MeV He+-implantations. The nx, ny, and nz refractive index modifications are studied. Losses in nonannealed YCOB waveguides measured with a CCD camera are found to be less than 2 dB cm(-1). This work is the first step toward the investigation of frequency conversion within the obtained guiding structures. PMID- 14735969 TI - Numerical studies on time-domain responses of on-off-keyed modulated optical signals through a dense fog. AB - We present a numerical technique to simulate the propagation characteristics of an on-off-keyed modulated optical signal through fog. The on-off-keyed modulated light (a square wave) is decomposed into a finite number of harmonic components, and a numerical solution for the vector radiative transfer equation is obtained for each harmonic that corresponds to the modulation frequency. With this method we study the distortion and the pulse spread in the received signal due to attenuation and scattering. We investigate the propagation characteristic of the modulated signal with different communication system parameters. This information can be used to study communication channel reliability. PMID- 14735970 TI - Liquid chromatographic enantioseparation of spin-labelled beta-amino acids. AB - Direct and indirect high-performance liquid chromatographic (HPLC) methods were developed for the enantioseparation of spin-labelled, cyclic, chiral beta-amino acids containing nitroxide free radicals, trans-3-amino- 1-oxyl-2,2,5,5 tetramethylpyrrolidine-4-carboxylic acid (trans-POAC), cis-4-amino-1-oxyl-2,2,6,6 tetramethylpiperidine-3-carboxylic acid (cis-beta-TOAC) and their N-Fmoc protected analogues, synthesized in racemic and enantiomerically pure forms. The direct method involved the use of a Chiralcel OD-RH column, while indirect separation was carried out by application of either 2,3,4,6-tetra-O-acetyl-beta-D glucopyranosyl isothiocyanate or (S)-N-(4-nitrophenoxycarbonyl)-phenylalanine methoxyethyl ester as chiral derivatizing agent. Use of 1-fluoro-2,4 dinitrophenyl-5-L-alanine amide (Marfey's reagent) as chiral derivatizing agent failed because of the low of yield of the derivatization reaction. Selection and variation of the mobile phase was restricted by the sensitivity of the spin labelled amino acids to acidic conditions. Conditions affording the best resolution were found and the differences in separation capability of the methods were noted. The sequence of elution of the enantiomers was determined by different methods and, in the case of the beta-TOAC analogues, the absolute configurations of the enantiomers corresponding to each peak were identified. PMID- 14735971 TI - Isolation of poly(propylene glycol)s from water for quantitative analysis by reversed-phase liquid chromatography. AB - Procedures for the isolation of poly(propylene glycol)s (PPGs) from a water matrix have been developed. Solid-phase extraction with an octadecylsilica cartridge and elution with methanol or with a graphitised carbon black cartridge and elution with a mixture of dichloromethane-methanol (4:1) or liquid-liquid extraction with chloroform were all suitable for model samples. However, only liquid-liquid extraction was suitable both for model and real environmental samples. Methods for reversed-phase liquid chromatographic determination of PPGs based on derivatisation and ultraviolet or fluorescence detection have been developed. Four derivatisation agents [3,5-dinitrobenzoyl chloride, phenyl isocyanate, 1-naphthoyl chloride and 1-naphthyl isocyanate (NIC)] were tested. Only NIC was found to give good reproducibility as well as a satisfactory detection limit. Finally, a method with liquid-liquid extraction with chloroform, derivatisation with NIC and liquid chromatographic separation with fluorescence detection was established. The developed method shows a highly correlated linearity of the analytical signals of particular homologues within a wide concentration range (approximately from 0.01 to 10 mg l(-1)). The precision of measurements is satisfactory for homologues having 5-9 oxypropylene subunits and becomes worse with an increase in the number of oxypropylene subunits. The limit of detection is 2 microg l(-1) for the majority of homologues. The method is suitable for the isolation and quantitative determination of PPGs in river water samples and as a tool for biodegradation testing. PMID- 14735972 TI - Retention of 3,5-dinitrobenzoyl derivatives of linear alcohol polyethoxylates in reversed-phase liquid chromatographic system with acetonitrile-water mobile phase. AB - The influence of mobile phase composition (acetonitrile-water ratio) on the separation of derivatised linear alkyl polyethoxylates (LAEs) is evaluated using thermodynamic quantities (Gibbs energy, enthalpy and entropy). In comparison to homologue series of alcohols oligomers of LAEs show irregular chromatographic behaviour that is demonstrated in irregular changes of thermodynamic quantities. It might be explained considering an influence of some of the following processes or their combinations on the retention of LAEs. These processes are solvation of oxyethylene chains in mobile phase, their interaction with silanols on silica surface of stationary phase and possibly their conformation changes. The composition of a mobile phase affects the mentioned processes and that is why the retention of LAEs is strongly (for the reversed-phase system unusually) sensitive to this composition in the studied range (volume fraction of acetonitrile phi = 0.90, 0.96 and 0.98). The experimental data also support the idea of the active role of stationary phase in the reversed-phase system. PMID- 14735973 TI - Reproducibility of low and high concentration data in reversed-phase liquid chromatography. II. Overloaded band profiles on Chromolith-C18. AB - Single-component adsorption isotherm data were acquired by frontal analysis (FA) for six low molecular weight compounds (phenol, aniline, caffeine, o-toluidine, p toluidine and propylbenzoate) on one Chromolith-C18 column (#30, Merck, Darmstadt, Germany), using different methanol:water solutions (composition between 60/40 and 15/85 v/v, depending on the solute) as the mobile phase. These data were modeled for best agreement between the experimental data points and the adsorption isotherm model. The adsorption-energy distributions were also derived and used for the selection of the best isotherm model. Widely different models were obtained for these six compounds, four being convex upward (i.e., Langmuirian) and two having at least one inflection point. Overloaded band profiles corresponding to two different sample sizes (a low and a high loading factor) were recorded on six monolithic columns (#30-35) belonging to the same manufactured lot. These experimental band profiles were compared to the profiles calculated from the isotherm measured by FA on the first column, using the equilibrium-dispersive (ED) model of chromatography. For four of the six columns (#30, #32, #33, and #35), the reproducibility was better than 5 and 2.5% for the low and the high concentration profiles, respectively. On the other two columns (#31 and #34), the bands showed significant and systematic retention time shifts for all six compounds (with nearly identical band shapes), the relative adsorption being between 6 and 15% stronger on column #31 or between 2 and 7% lower on column #34. These differences seem to be correlated with the differences in the total porosities of these columns, which differ by 3% from columns #31 to #34, the higher porosity column giving the stronger adsorption. PMID- 14735974 TI - Split flow and bypass flow systems for monolithic capillary columns in liquid chromatography. AB - Split flow and bypass flow systems were assembled using Nano Y Connectors with low dead volume commercially available for capillary liquid chromatography (LC). The split ratio could be controlled by changing the dimension of restriction tubing and applied back pressure to the restriction tubing. The split flow system allowed us to use valve injectors and pumps commercially available for capillary LC. The reproducibility of the present split flow system was acceptable. The relative standard deviation for six successive measurements was 0.4% for the retention time, whereas that for the peak height and peak area was 1-3% depending on the analytes. The bypass flow system uses two Nano Y Connectors, where the eluent split at the first Nano Y Connector, which is located in the inlet of the separation column, is merged again into the effluent from the column at the second Nano Y Connector. The bypass flow system could avoid on-column detection and allowed us to use flow cells, leading to an approximate three times improvement in signal-to-noise. The present flow systems were evaluated by using aromatic hydrocarbons and alkylbenzenes as test analytes. PMID- 14735975 TI - Nuclear magnetic resonance analysis of silica gel surfaces modified with mixed, amine-containing ligands. AB - Different approaches for quantitative analysis by 29Si and 13C CP/MAS nuclear magnetic resonance (NMR) of silica gel chemically modified by a mixture of long and short chain amines, -(O)3Si(CH2)3N(CH3)2(CH2)13CH3 and -(O)3Si(CH2)3N(CH3)3, are compared to elemental analysis. Unlike 29Si NMR, variable contact time data are necessary for accurate quantitative analysis by 13C NMR. Surprisingly, spectral overlap does not interfere with this approach. Surfaces prepared from reaction mixtures that consisted of 67 and 33% (v/v) long chain are found to actually contain 37 and 16% long chain amines, respectively. The mixed phase surfaces have more extensive cross-linking and fewer unreacted hydroxyls than single phase surfaces. PMID- 14735976 TI - Characterization of calixarene- and resorcinarene-bonded stationary phases. I. Hydrophobic interactions. AB - New HPLC phases with supramolecular selectors on the basis of calixarenes and resorcinarenes were investigated for the first time by means of empirically based test mixtures. The tests, originally developed for common reversed phases, were chosen to evaluate fundamental chromatographic properties of the new materials. In the first part of these studies three descriptors (hydrophobic retention capacity--k'(hyd), hydrophobic selectivity--alpha(hyd), steric selectivity- alpha(ster)) were determined. Except of higher alpha(ster) values and alpha(hyd) values with some methods for the resorcinarene phase the phases with supramolecular selectors were classified as less hydrophobic possessing lower hydrophobic and steric selectivities compared to three RP-C18 phases and a p-tert butyl phenyl ether phase. The results were confirmed by means of a separation of geometric isomers of thioxanthenes. In contrast, in spite of lower k'(hyd) and alpha(hyd) values calixarene phases were more selective than the Kromasil-C18 phase in the separation of gestagenic and androgenic steroids due to specific interactions with the steroids of similar lipophilicity. PMID- 14735977 TI - High-performance liquid chromatographic enantioseparation using chitin carbamate derivatives as chiral stationary phases. AB - Chitin carbamate derivatives including 4-substituted and 3,5-disubstituted phenylcarbamates, 1-phenylethylcarbamates, and cycloalkylcarbamates were synthesized and coated on macroporous silica gel to evaluate their chiral recognition abilities as chiral stationary phases (CSPs) for high-performance liquid chromatography (HPLC). Among the derivatives, the 3,5-dimethylphenyl, 4 chlorophenyl, and 4-trifluoromethylphenylcarbamates showed relatively high-chiral recognition abilities when a hexane-2-propanol mixture was used as the eluent. The CSPs based on the chitin 3,5-dimethylphenyl and 3,5-dichlorophenylcarbamates could be stably used in the presence of chloroform and ethyl acetate as a component of the eluents, and a few racemates were more sufficiently resolved by the addition of a small amount of chloroform in the mobile phase. Some racemates were more efficiently resolved under the reversed phase condition. PMID- 14735979 TI - Mechanism and kinetics of protein transport in chromatographic media studied by confocal laser scanning microscopy. Part II. Impact on chromatographic separations. AB - The impact of different transport mechanism on chromatographic performance was studied by confocal laser scanning microscopy (CLSM) for solutions containing bovine serum albumin (BSA) and monoclonal IgG 2a under different solid- and fluid phase conditions. During this investigation, a clear influence of the uptake mechanism on the affinity of the respective proteins for the different adsorbents and thus separation performance of the chromatographic process could be observed. For the system SP Sepharose Fast Flow at pH 4.5 pore diffusion could be ascribed to be the dominant transport mechanism for both proteins and the adsorption profiles resembled a pattern similar to that described by the 'shrinking core' model. Under these conditions a significantly higher affinity towards the adsorbent was found for BSA when compared to IgG 2a. With changing fluid- and solid-phase conditions, however, a change of the transport mode for IgG 2a could be detected. While the exact mechanism is still unresolved it could be concluded that both occurrence and magnitude of the now governing transport mechanism depended on protein properties and interaction with the adsorbent surface. For the system SP Sepharose XL at pH 5.0 both parameters leading to the change in IgG 2a uptake were combined resulting in a clear change of the system affinity towards the IgG 2a molecule, while BSA adsorption was restricted to the most outer shell of the sorbent. PMID- 14735978 TI - Mechanism and kinetics of protein transport in chromatographic media studied by confocal laser scanning microscopy. Part I. The interplay of sorbent structure and fluid phase conditions. AB - An experimental study on the interplay of sorbent structure and fluid phase conditions (pH) has been carried out examining adsorption and transport of bovine serum albumin (BSA) and a monoclonal antibody (IgG 2a) on SP Sepharose Fast Flow and SP Sepharose XL. SP Sepharose Fast Flow is characterised by a relatively open pore network, while SP Sepharose XL is a composite structure with ligand-carrying dextran chains filling the pore space. Both adsorbents have similar ionic capacity. Protein transport and adsorption profiles were evaluated using confocal laser scanning microscopy. Under all investigated conditions, BSA uptake could be adequately explained by a pore diffusion mechanism. The adsorption profiles obtained for IgG 2a, however, indicated that changes in fluid phase conditions as well as a change in the solid phase structure could result in a more complex uptake mechanism as compared to pore diffusion alone. This mechanism results in a fast transport of proteins into the adsorbent, followed by an overshoot of protein in the center of the sorbent and a setback towards a homogeneous adsorption profile. PMID- 14735980 TI - Supercritical fluid extraction of grape seed oil and subsequent separation of free fatty acids by high-speed counter-current chromatography. AB - Supercritical fluid extraction (SFE) of grape seed oil was performed to study the effect of various parameters such as pressure, temperature and the particle size of the sample on the yield and composition of oil using an analytical-scale SFE system. Then the extraction was scaled up by 125 times using a preparative SFE system under the optimized conditions of high pressure (30-40 MPa) and low temperature (35-40 degrees C) with medium particle size (20-40 mesh). The maximum yield of the oil can reach 6.2% with pure supercritical CO2 and 4.0% more oil can be obtained by adding 10% of ethanol as modifier. The unsaturated fatty acids (UFSs) make up about 70% in the oil on the basis of free fatty acids. The grape seed oil was then subjected to separation and purification for free fatty acids after saponification by high-speed counter-current chromatography coupled with evaporative light scattering detection (ELSD). The separation of 1.0 g of oil can yield about 430 mg pure linoleic acid at 99% purity. The fatty acids were analyzed by HPLC-ELSD. PMID- 14735981 TI - Liquid chromatographic determination of geometrical retinol isomers and carotene in enteral feeding formulas. AB - The objective of the present study was to evaluate a practical method for separation and determination of geometrical retinol isomers and carotene in enteral feeding formulas and to analyze 17 samples of commercial formulas. By using a normal-phase HPLC column and a mobile phase consisting of 1-octanol in n hexane, seven isomers of retinol were separated and identified from the standard solution after photolysis. For evaluation of Vitamin A activity in these formulas, simultaneous determination of total carotene was performed. The data about linearity, recovery, accuracy and precision showed the reliability of analytical procedures. In the unsaponifiable portion of samples of commercial formulas, six retinol isomers were identified: (E)-retinol; (Z)-13; (Z)-9; (Z,Z) 9,13; (Z,Z)-11,13 and (Z)-7-retinol. (Z)-13/(E)-retinol ratios ranged between 3 and 37%. The range of total Z/(E)-retinol isomers fell between 5 and 42%. Despite the high concentration of Z-isomers observed in various commercial enteral feeding formulas, none of the samples presented Vitamin A activity below 90% of that specified on the label. PMID- 14735982 TI - Determination of acidic pharmaceuticals, antibiotics and ivermectin in river sediment using liquid chromatography-tandem mass spectrometry. AB - Analytical methods have been developed for the determination of eight acidic pharmaceuticals and two metabolites, seven antibiotics and the parasiticide ivermectin in a selected river sediment. The sediments were solvent extracted with ultrasonic assistance. A solid phase extraction (SPE) clean-up step was performed thereafter. The acidic compounds clofibric acid, diclofenac, fenoprofen, gemfibrozil, ibuprofen, 2-hydroxy-ibuprofen, indomethacin, ketoprofen, naproxen and the parasiticide ivermectin were measured in the negative mode by LC-APCI-tandem MS, whereas the antibiotics clarithromycin, erythromycin, roxithromycin, sulfadiazine, sulfamethazine, sulfamethoxazole and trimethoprim were detected in the positive mode by LC-ESI-tandem MS. Bezafibrate could not be determined in the sediment using the method developed. The limit of quantification (LOQ) ranged from 0.4 to 8 ng g(-1) for the acidic pharmaceuticals, sulfadiazine and ivermectin and was 20 ng g(-1) for the other antibiotics. PMID- 14735983 TI - Application of carbohydrate analysis to verify honey authenticity. AB - Gas chromatography and liquid chromatography have been used simultaneously to analyze sugars in honey. After statistical processing by principal components analysis, additions of exogenous sugars could be detected by the appropriate fingerprints of adulteration. Application to acacia, chestnut and lavender honeys enabled the detection of fraud resulting from 5 to 10% addition of sugar syrups. This method may be considered as a replacement of isotopic analysis, that has some limitations. PMID- 14735984 TI - Reevaluation of headspace solid-phase microextraction and gas chromatography-mass spectrometry for the determination of methyl tert-butyl ether in water samples. AB - A headspace solid-phase microextraction (SPME) and gas chromatography-mass spectrometry (GC-MS) method has been reevaluated for the determination of methyl tert-butyl ether (MTBE) in water. Factors affecting method performance were examined. The internal standard, deuterated MTBE, is necessary because it allows normalization of the calibration since the deuterated MTBE and MTBE experience the same extraction efficiency. The method detection limit was 6.6 ng/l, with a linear range up to 160 microg/l. SPME extraction efficiency and detection sensitivity for MTBE decreased in the presence of gasoline matrix. However, reliable measurement of MTBE was possible in the background matrix concentration of 0.97 mg/l gasoline using deuterated internal standard. Reproducibility was 4.5% at the spiking level of 47 ng/l. The average recoveries of MTBE in spiked fountain water, tap water, and snow at 60 ng/l and in spiked rainwater at 580 ng/l ranged from 101 to 105%. The method was successfully applied to the analysis of trace levels of MTBE in various water samples, including rain and snow samples collected in Albany, NY, USA. PMID- 14735985 TI - Determination of 39 polybrominated diphenyl ether congeners in sediment samples using fast selective pressurized liquid extraction and purification. AB - In order to reduce time of analysis, a new pressurized liquid extraction (PLE) method that automatically and rapidly achieves quantitative and selective extraction of 39 polybrominated diphenyl ether (PBDE) congeners in sediment samples was optimized. It consists of on-line cleanup by inclusion of sorbents in the extraction cell. The new method was compared with a conventional method based on the use of Soxhlet extraction followed by solid-phase extraction (SPE) with cartridges. The instrumental determination was performed by GC-MS, using negative chemical ionization in the selected-ion monitoring mode. Recoveries from 47 to 82% were obtained for spiked tri- to hepta-PBDE congeners in sediment sample. The repeatability of replicate extractions was better than 15% relative standard deviation. The detection limits obtained with the new developed method were between 1 and 46 pg/g dry weight. The reduction in the sample preparation (extraction + cleanup) time (from days to 30 min) with a similar efficiency than that afforded by the conventional Soxhlet extraction-SPE cleanup technique indicates the suitability of this method. The method was applied to sediment samples where the analytes were detected in the range of 0.86-2.49 ng/g dry mass. PMID- 14735986 TI - Determination of aliphatic amines using N-succinimidyl benzoate as a new derivatization reagent in gas chromatography combined with solid-phase microextraction. AB - A simple, selective and sensitive approach was developed for the quantitation of aliphatic amines in lake water applying a new reagent (N-succinimidyl benzoate, SIBA), synthesized in the laboratory of the authors. Derivatization of the n-C1 C6 aliphatic monoamines and dimethylamine in aqueous solution with SIBA was followed by headspace solid-phase microextraction (SPME). Derivatives were identified by gas chromatography-mass spectrometry and determined by gas chromatography-flame ionization detection. Both derivatization and SPME conditions have been optimized. Derivatizations were performed in borate buffer (pH 8.8), at 60 degrees C for 22 min. SPME was carried out from saturated sodium chloride solution, at 80 degrees C for 60 min, desorption at 250 degrees C for 2 min. Detection limit of derivatized amines proved to be 0.13-7.2 nmol/l, while recovery of amines from lake water samples, in the concentration range of 100-200 microg/l, varied from 94.1 to 102.7%. PMID- 14735987 TI - Studies on a poly(styrene-divinylbenzene)-encapsulated sub-micrometer sized organic-inorganic hybrid silica packing, its synthesis, some of its characteristics and applications. AB - The hydrolyzed product of a mixture of tetraethoxysilane (TEOS) and vinyltriethoxysilane (VTEOS) was used to produce organic-inorganic hybrid silica by sol-gel processing. The product was a kind of composite with vinyl groups incorporated into the structure of the final particles through C-Si bonds. These hybrid silica particles were spherical and the average diameter was about 350 nm as measured by scanning electron microscopy (SEM). The particles were non-porous as proved by BET method. The hybrid silica particles were subsequently encapsulated with a layer of PS-DVB, a polymeric mixture of styrene and divinylbenzene, with the vinyl groups on the particle surface as anchors of the polymer chains. The encapsulated packing was used as stationary phases in CEC, and the chromatographic behaviors were studied. Experimental results showed that, as expected, the smaller diameter stationary phase resulted in higher column efficiencies. In comparison with the columns packed with 5 microm diameter encapsulated-silica stationary phases of the same kind and other sub-micron stationary phases reported, it can be concluded that the EOF velocity is independent of the particle diameter when using 350 nm phase, but determined by the surface nature of the stationary phase. PMID- 14735988 TI - Chiral separation of fluorescamine-labeled amino acids using microfabricated capillary electrophoresis devices for extraterrestrial exploration. AB - Chiral separations of fluorescamine-labeled amino acids are characterized and optimized on a microfabricated capillary electrophoresis (CE) device. A standard mixture of acidic and neutral amino acids is labeled with fluorescamine in less than 5 min and the hydroxypropyl-beta-cyclodextrin (HPbetaCD) concentration, temperature, and pH are optimized (15 mM HPbetaCD, 6 degrees C, pH < 9) to achieve high-quality and low background chiral separations in less than 200 s. All four stereoisomers formed in the labeling reaction of the chiral dye with the chiral amino acids are typically resolved. At pH > 9, isomerization of the dye chiral center is observed that occurs on the time scale of the chip separation. Typical limits of detection are approximately 50 nM. These results demonstrate the feasibility of combining fluorescamine labeling of amino acids with microfabricated CE devices to develop low-volume, high-sensitivity apparatus and methods for extraterrestrial exploration. PMID- 14735989 TI - Subsecond separation of cellular flavin coenzymes by microchip capillary electrophoresis with laser-induced fluorescence detection. AB - In this article, it was demonstrated that a subsecond separation of cellular metabolites such as riboflavin (RF), flavin mononucleotide (FMN), and flavin adenine dinucleotide (FAD) was achieved using microchip capillary electrophoresis with laser-induced fluorescence detection. The influences of crucial parameters that governed analysis time (e.g., channel length and electric field for separation) and separation resolution (e.g., sample size) were investigated, both in theoretical aspects and experimental practice. Quantitative analyses were performed that exhibited linear dynamic range of two orders of magnitude, with calculated detection limits of 34, 201, and 127 nM for RF, FAD, and FMN, respectively. To test the validity of the method, it was successfully applied to characterize several recombinant flavin-binding domains in a human neuronal nitric oxide synthase. PMID- 14735990 TI - Determination of abscisic acid by capillary electrophoresis with laser-induced fluorescence detection. AB - A novel method based on capillary electrophoresis coupled to laser-induced fluorescence detection (CE-LIF) was developed for the determination of abscisic acid (ABA), which is an essential phytohormone during plant growth and development. ABA was labeled with 8-aminopyrene-1,3,6-trisulfonate via reductive amination in presence of acetic acid and sodium cyanoborohydride. The derivatization yield was maximized by optimizing several derivatization parameters including derivatization reagent concentration, reaction temperature and time. The conjugate was separated and quantitated by CE-LIF. The linearity of ABA was determined in the range from 0.1 to 10 micromol l(-1) with a correlation of 0.9979. The derivatization limit of detection for ABA was found to be 56 fmol (corresponding to the concentration of 2.8 x 10(-8) mol l(-1)). The detection limit for ABA was 5.5 amol for an injection volume of 5 nl. As a preliminary application, the proposed method was successfully applied to determining trace amount of ABA in the crude extracts of tobacco without extra purification and enrichment procedure and showed a better selectivity and sensitivity than those conventional methods used in determination of ABA. PMID- 14735991 TI - Identifying research needs for TB vaccines. PMID- 14735992 TI - Evaluation of the basal and post-adrenocorticotrophic hormone serum concentrations of 17-hydroxyprogesterone for the diagnosis of hyperadrenocorticism in dogs. AB - Serum concentrations of 17-hydroxyprogesterone and cortisol were measured before and after the administration of exogenous adrenocorticotrophic hormone (ACTH) to three groups of dogs: 27 healthy dogs (group 1), 19 dogs with non-adrenal illness (group 2) and 46 dogs with hyperadrenocorticism (group 3). The median (range) post-ACTH concentrations of 17-hydroxyprogesterone were 5.0 (22.2 to 16.8), 6.9 (2.0 to 36.2) and 14.4 (1.7 to 71) nmol/litre in groups 1, 2 and 3, respectively. There were no significant differences in the basal or post-ACTH concentrations of cortisol or 17-hydroxyprogesterone between groups 1 and 2. The post-ACTH concentrations of 17-hydroxyprogesterone in group 3 were significantly (P<0.001) greater than those in groups 1 and 2 combined. The area under the receiver operating curve (ROC) for the post-ACTH concentration of cortisol (0.94) was significantly greater than that for the post-ACTH concentration of 17 hydroxyprogesterone (0.76). Using a two-graph ROC analysis, a cut-off of 8.5 nmol/litre was found to maximise both the sensitivity and specificity of the post ACTH concentration of 17-hydroxyprogesterone for the diagnosis of hyperadrenocorticism at 71 per cent. With a cut-off of 4.5 nmol/litre the sensitivity increased to 90 per cent but the specificity decreased to 40 per cent; with a cut-off of 16.7 nmol/litre the specificity increased to 90 per cent but the sensitivity decreased to 47 per cent. PMID- 14735993 TI - Outbreaks of adenoviral gizzard erosion in slaughtered broiler chickens in Japan. AB - Gizzard erosion in broiler chickens was investigated at 18 slaughterhouses in Japan. The condition was observed in 13 of them, and adenoviral gizzard erosion (AGE) was diagnosed histologically, immunohistochemically and virologically in the eroded gizzards from nine of these 13. The antigen-positive intranuclear inclusion body of group 1 fowl adenovirus was observed in the epithelial cells of the affected gizzards, and fowl adenoviruses were isolated from the lesions. In two of the slaughterhouses the total weights of the gizzards disposed of in three years were 3590 kg (0.40 per cent of the gizzards inspected) and 2880 kg (0.19 per cent). Sixteen of the 19 outbreaks of gizzard erosion on 15 farms that were confirmed in three of the slaughterhouses, including the previous two slaughterhouses, were diagnosed as AGE, and the condition was suspected in the other three outbreaks. Most of the adenoviruses isolated were identified as serotype-1 by PCR-restriction fragment length polymorphism. No apparent clinical signs were observed in any of the affected flocks. PMID- 14735994 TI - Congenital myasthenic syndrome of Brahman cattle in South Africa. AB - A congenital myasthenic syndrome in Brahman cattle is caused by a homozygous 20 base pair deletion (470del20) in the gene coding for the epsilon subunit of the acetylcholine receptor at the neuromuscular junction. It causes a progressive muscle weakness starting either at birth or within the first month. A PCR-based DNA test, using blood or semen stored on FTA paper, was developed and validated; the test makes it possible to differentiate rapidly and accurately between homozygous wild-type, heterozygous and homozygous affected animals. Preliminary testing of Brahman cattle in South Africa has revealed several carrier animals, some of them influential animals in the breeding population. PMID- 14735995 TI - Results of a postal survey in 2002 into the occurrence of scrapie in Great Britain. PMID- 14735996 TI - Mammary neoplasia with lung metastasis in a rat (Rattus norvegicus). PMID- 14735997 TI - Peripheral nerve sheath tumour in a cow with clinical signs consistent with vagal neuropathy. PMID- 14735998 TI - Urolithiosis in two lions. PMID- 14735999 TI - Use of random amplified polymorphic DNA analysis to verify antimicrobial efficacy when treating Staphylococcus aureus mastitis in cows. PMID- 14736000 TI - Human generic products and the prescribing cascade. PMID- 14736001 TI - Lay TB testing. PMID- 14736002 TI - TB breakdowns in closed herds. PMID- 14736003 TI - Gangrenous mastitis in dairy herds. PMID- 14736004 TI - Nephrology in the clinic and quality in nephrology. AB - The majority of patients with chronic kidney disease are followed-up in pre dialysis clinics for a varying length of time, often for several years. The renal team (physicians, nurses, dieticians, social assistants) intervenes on renal disease progression and educates the patient regarding the adjustments of his or her life-style to the limitations imposed by the disease. Pre-dialysis clinics are often overburdened and it is uncertain whether renal care should be based exclusively on renal teams, a difficult and costly option, or if it could be efficaciously organized by involving other health professionals. The Journal of Nephrology will maintain strong attention on these crucial issues for the future of nephrology. PMID- 14736005 TI - Renal replacement therapy in lupus nephritis. AB - The indications and the choice of renal replacement therapy for lupus patients are similar to those for other uremic patients. However, lupus patients can pose some particular problems. First, 10-28% of patients needing dialysis can have a partial renal function recovery. Therefore, the clinician has to decide whether to administer a rescue treatment, risking side-effects, or to reduce immunosuppression precluding a potential recovery. Many patients on regular dialysis show subdued biological and clinical activity. Others can show a hectic disease activity, particularly in the 1st year. In these cases, treatment is difficult, as vigorous immunosuppression can expose uremic patients to severe side-effects. The presence of circulating antiphospholipid antibodies (aPL) can favor thrombosis or stenosis of vascular access (VA). Renal transplantation is the best therapy for most lupus patients with end-stage renal failure. Many, but not all, studies have reported similar patient and graft survival rates in lupus and in non-lupus transplant recipients. The results are much better with living donor transplantation. Patients with aPL, black patients and those on long-term dialysis have a higher graft failure risk. Candidates with active lupus and/or those with significant iatrogenic morbidity should be advised to wait 6-12 months before transplantation. The recurrence risk of lupus nephritis ranged between 2% and 30% in different studies. The histological picture does not usually show severe features. Antiplatelet agents or anticoagulation can be advised for aPL patients. PMID- 14736006 TI - Pregnancy in kidney transplantation: satisfactory outcomes and harsh realities. AB - Since the first successful case of a pregnancy reported 40 yrs ago in a woman receiving a kidney transplant from her identical twin sister who did not receive immunosuppressive medications, the dream of a pregnancy in a renal transplant recipient has become reality. In women of childbearing age with a functioning transplant, the pregnancy rate has improved from 2 to 5%. Approximately 35% of pregnancies do not progress beyond the 1st trimester; the success rate is > 90% after the 1st trimester. In this review, different aspects of this topic are discussed: the consequences of pregnancy on renal grafts and maternal morbidity (hemodynamic changes, immunological problems, hypertension/preeclampsia, urinary tract infections and renal damage progression), the influence of renal grafts on pregnancy (perinatal mortality, prematurity, intrauterine growth retardation, low birth weight, malformations, handicaps and immunological problems) and the role of drugs used for renal transplants. A pregnancy can have a successful outcome if pre-conceptional graft function is good, if hypertension is absent and if the interval from grafting is at least 2 yrs. However, the majority of live-born outcomes are premature and many are low birth weight. Recipients must be advised that their offspring can also suffer from immunological abnormalities, malformations, long-term handicaps, and that the deleterious effects of pregnancy on long-term graft function cannot be excluded. In conclusion, women of childbearing age who have had renal transplantation should be counselled before conception about possibility and risks of pregnancy. PMID- 14736008 TI - Epidemiology of end-stage renal disease in an interregional perspective: Registries of Puglia and Basilicata, southern Italy. AB - BACKGROUND: This report on the 1994-1998 Dialysis and Transplantation Registry (DTR) of Puglia and Basilicata provides the first epidemiological profile of ESRD in southern Italy. METHODS: Frequency measures of patients in renal replacement therapy (RRT) were computed for Puglia and Basilicata (inhabitants: 4,086,422 and 610,000 respectively). Hazard ratios (HR) of death in relation to sex, age, educational level, primary nephropathies, and modality of dialysis, were estimated by applying the Cox model to patients starting dialysis as first RRT in 1994-1998 in Puglia. RESULTS: The prevalence of treated ESRD in Puglia was 881 per million population (p.m.p.) (dialysis: 721 p.m.p.) in 1998, 713 p.m.p. (dialysis: 617 p.m.p.) in 1994. In Basilicata the prevalence of ESRD was 795 p.m.p. (dialysis: 669 p.m.p.) in 1998, 636 p.m.p. (dialysis: 575 p.m.p.) in 1994. Mean age at start of dialysis of incident cases of Puglia was 60 yr (median: 64 yr). Figures of diabetes, vascular diseases, and glomerulonephritis, were: 16%, 21%, 17%. Out of 2,152 incident patients on dialysis for at least one month, 293 started with peritoneal dialysis (PD). A 60-70% higher risk of death was observed for diabetic nephropathy and PD. In the Puglia/Basilicata DTR pooled analysis, lower educational level was associated with a 60% increased mortality risk. CONCLUSIONS: The associations of PD and low education with the risk of death are very likely to be due to comorbid conditions, unavailable in these databases as in most regional and national DTR. By looking at variations of rates and outcomes among areas, potential improvements of local DTR for planning and research uses are discussed. PMID- 14736007 TI - The benefit of STent placement and blood pressure and lipid-lowering for the prevention of progression of renal dysfunction caused by Atherosclerotic ostial stenosis of the Renal artery. The STAR-study: rationale and study design. AB - BACKGROUND: Atherosclerotic renal artery stenosis (ARAS) is associated with progressive loss of renal function and is one of the most important causes of renal failure in the elderly. Current treatment includes restoration of the renal arterial lumen by endovascular stent placement. However, this treatment only affects damage caused by ARAS due to the stenosis and ensuing post-stenotic ischemia. ARAS patients have severe general vascular disease. Atherosclerosis and hypertension can also damage the kidney parenchyma causing renal failure. Medical treatment focuses on the latter. Lipid-lowering drugs (statins) could reduce renal failure progression and could reduce the overall high cardiovascular risk. The additional effect on preserving renal function of stent placement as compared to medical therapy alone is unknown. Therefore, the STAR-study aims to compare the effects of renal artery stent placement together with medication vs. medication alone on renal function in ARAS patients. METHOD: Patients with an ARAS of > or = 50% and renal failure (creatinine (Cr) clearance < 80 mL/min/1.73 m2) are randomly assigned to stent placement with medication or to medication alone. Medication consists of statins, anti-hypertensive drugs and antiplatelet therapy. Patients are followed for 2 yrs with extended follow-up to 5 yrs. The primary outcome of this study is a reduction in Cr clearance > 20% compared to baseline. This trial will include 140 patients. PMID- 14736009 TI - Arteriovenous fistula use and heart disease in long-term elderly hemodialysis patients: analysis of United States Renal Data System Dialysis Morbidity and Mortality Wave II. AB - BACKGROUND: There are some concerns that arteriovenous fistula (AVF) use or other dialysis specific factors may exacerbate cardiovascular disease in long-term hemodialysis (HD) patients. METHODS: We performed a historical cohort study of the United States Renal Data System Dialysis Morbidity and Mortality Wave II study, limited to 993 patients who started HD in 1996 with valid information on vascular access and who were primarily eligible for Medicare at the start of the study. We assessed the association between hemodialysis vascular access and heart disease, defined as Medicare Claims for heart failure (HF, International Classification of Diseases (ICD9) code 428.x) and acute coronary syndromes (ACS, ICD9 code 410.x and 411.x). Cox proportional hazards regression (using propensity analysis) was used to model adjusted hazard ratios (AHR) for the association between patients factors and heart disease after dialysis. RESULTS: The rate of HF per 100 person years at risk (PYAR) was 19.6 among AVF users, 25.7 among patients using polytetrafluoroethylene grafts (grafts), and 31.1 among patients using temporary catheters. Corresponding rates of ACS were 8.2 among AVF users, 11.0 among users of grafts, and 12.4 among users of temporary catheters. In Cox Regression analysis, there was no significant association between AVF use and either HF or ACS. This lack of association was consistent across gender, diabetes, race, and age. CONCLUSIONS: We found that AVF use had no significant association with the incidence of HF or ACS. We conclude that use of AVF by 60 days after the start of dialysis is not associated with an increased risk of later non-fatal cardiovascular outcomes in long-term hemodialysis patients. PMID- 14736010 TI - Proteinase 3/myeloblastin as a growth factor in human kidney cells. AB - BACKGROUND: Numerous data support the possible role of myeloblastin/proteinase 3 (PR3) in growth and differentiation of neutrophil granulocytes and certain monocyte subtypes. However, whether PR3 is expressed in non-myeloid cells remains a matter of debate even though recent studies clearly demonstrated its expression in endothelium, kidney epithelial cells and epithelial tumor cell lines. METHODS: To survey PR3 transcript presence in human tissues, we analyzed different human tissues by dot blot and northern blot using a cloned PR3-cDNA probe. To examine the physiological function of PR3 expression in non-myeloid cells, we constructed different recombinant retroviral vectors containing human PR3-cDNA variants and expressed them in tubular epithelial cells (TEC). Using an MTT-based proliferation assay, we determined the proliferation rate of PR3-transduced kidney cells. RESULTS: The resulting expression pattern clearly indicated that PR3 transcripts are not only present in tissues known to harbor hematopoietic cells, but surprisingly PR3 was highly expressed in fetal organs including the kidney. The proliferation assay revealed that the growth rate of TEC transduced with native PR3 was significantly enhanced relative to non-transduced TEC. CONCLUSIONS: The results supported our theory that PR3 can act as a growth factor in non-hematopoietic cells, analogous to its role in hematopoietic cells. The cells, recombinant vectors and methods described here serve as a basis to investigate PR3 function in cellular differentiation and proliferation, as well as its role in autoimmune diseases. PMID- 14736011 TI - Effects of candesartan, an angiotensin II type 1 receptor blocker, on diabetic nephropathy in KK/Ta mice. AB - BACKGROUND: Although therapeutic effects of angiotensin II type 1 receptor blocker (ARB) on renal injury in non-insulin dependant diabetes mellitus (NIDDM) have been demonstrated, the beneficial effects and their mechanisms in diabetic nephropathy have not been well evaluated. METHODS: KK/Ta mice were divided into three groups according to the treatment: candesartan 4 mg/kg/day from 6 to 28 weeks of age (group I; early treatment); from 12 to 28 weeks of age (group II; late treatment); only vehicle (group III). BALB/c mice treated with vehicle were used as controls (group IV). Body weight (BW), systolic blood pressure (SBP), blood glucose, urinary type IV collagen and albumin excretion were measured every 4 weeks. Morphometry and immunohistology of albumin, transforming growth factor beta1 (TGF-beta1) and Smad7 were performed in all groups. RESULTS: BW and blood glucose were higher in groups I, II and III than in group IV from 8 weeks. SBP was markedly reduced in groups I and II compared with group III (p < 0.05, p < 0.005). Urinary type IV collagen and albumin excretion were increased in group III compared to group IV (p < 0.05, p < 0.005), whereas they were reduced in groups I and II when compared to group III (p < 0.05). Morphometric analysis revealed that the whole glomerular area (WGA), glomerular tuft area (GTA), extracellular matrix area (ECMA) and intraglomerular cell nuclei number (NIGCN) were significantly reduced in groups I, II and IV compared to group III at 28 weeks. In immunohistochemistry, TGF-beta1 expression in both glomeruli and tubules of groups I and II decreased compared to that of group III at 28 weeks, while Smad7 in group III glomeruli was reduced compared to that in groups I and II. CONCLUSIONS: It appears that candesartan reduced urinary type IV collagen and albumin excretion, and attenuated glomerular hypertrophy and mesangial matrix accumulation by the TGF-betaS/Smad signaling pathway in KK/Ta mice with diabetic nephropathy. PMID- 14736012 TI - Cholesterol metabolism in patients with chronic renal failure on hemodialysis. AB - Premature atherosclerosis is a major concern in patients on chronic dialysis and the identification of risk factors is important for preventive and interventional strategies. Other than the recognized atherogenic lipoprotein levels, little is known about overall cholesterol metabolism in patients on chronic hemodialysis (HD) and the best therapeutic intervention is still being debated. Therefore, we investigated intestinal cholesterol absorption, cholesterol and bile acid synthesis, and non-cholesterol plasma sterols in eight patients on dialysis and compared the results to those of 16 healthy male controls matched for body mass index and dietary cholesterol intake. Total, low-density lipoprotein (LDL) cholesterol, and triglycerides did not differ between the groups, but dialysis patients had a significantly lower high-density lipoprotein (HDL) cholesterol level (39 +/- 11 mg/dL vs. 48 +/- 10 mg/dL, p < 0.045). However, fractional cholesterol absorption, was significantly lower in dialysis patients (42.8 +/- 10.9% vs. 53.4 +/- 11%, p < 0.035), whereas plasma plant sterol concentrations and their ratios to cholesterol did not differ. Bile acid and total cholesterol synthesis were lower in dialysis patients (40% and -25%, respectively), although the differences were not significant. In contrast, lathosterol and its ratio to cholesterol in plasma was significantly lower in dialysis patients (0.176 +/- 0.084 mg/dL vs. 0.251 +/- 0.102 mg/dL, p < 0.024 and 0.733 +/- 0.353 microg/mg vs. 1.172 +/- 0.407 microg/mg, p < 0.017, respectively), indicating reduced hepatic de novo cholesterol synthesis. It is concluded that reduced HDL cholesterol and reduced bile acid synthesis contributes to atherosclerosis pathogenesis in dialysis patients, whereas intestinal cholesterol absorption and hepatic cholesterol synthesis did not seem dominant in this process at this stage of disease. Consequently, treatment with bile acid binding resins could be preferable to treatment with cholesterol absorption and synthesis inhibitors. PMID- 14736013 TI - Late referral defined by renal function: association with morbidity and mortality. AB - BACKGROUND: Many patients with chronic renal failure are referred very late to nephrology units. Late referral (LR) is reported to be associated with increased morbidity and mortality. METHODS: We used glomerular filtration rate (GFR) at the first visit to a nephrologist to define early referral (ER) and LR in a retrospective analysis. Patients admitted with a GFR < 20 mL/min/1.73 m2 were classified as LR. The 75 patients with chronic renal failure beginning renal replacement therapy (RRT) at Innsbruck University Hospital between January 1999 and October 2000 were included. Patient characteristics were compared between the two groups. Survival analysis until the end of 2002 was carried out using Cox's proportional hazard model. To identify the influence of comorbidity on mortality a comorbidity score was applied. RESULTS: Thirty-three patients were classified as ER and 42 patients as LR. Diabetic nephropathy was more frequent in the LR group (18 vs. 6 patients, p = 0.005). ER patients were significantly younger (53 +/- 16 yrs) as compared to LR patients (62 +/- 14 yrs, p = 0.012). Comorbid conditions were more frequent in the LR group (comorbidity score 1.5 +/- 1.3 for LR and 0.7 +/- 1.1 for ER, p = 0.003). During follow-up, 27 patients died, 19 from the LR group and 8 from the ER group. In the univariate analysis, comorbidity score (p < 0.001) and age (p = 0.017) were significantly associated with mortality, whereas LR patients demonstrated higher mortality (p = 0.076). By multivariate analysis the comorbidity score (p < 0.001) only was associated with mortality within at least 2 yrs of RRT. CONCLUSION: Over half of the patients with end-stage renal disease (ESRD) were referred too late, with a GFR < 20 mL/min/1.73 m2. Mortality during the 1st 2 yrs on RRT was mainly determined by comorbidity, acquired during the course of chronic renal failure. In comparison, the negative impact of LR seems to be minor and requires a larger sample size to be demonstrated. PMID- 14736014 TI - Structured pre-dialysis programs: more than just timely referral? AB - BACKGROUND: Patients who begin chronic dialysis within 3 months of seeing a nephrologist (late referral)have increased morbidity and mortality. However, it is not known whether formal pre-dialysis education programs (PEPs) are more advantageous, regarding early dialysis outcomes and resource use, than unstructured specialist follow-up. METHODS: We prospectively collected information on specialist care, unstructured or PEP, modality choice and hospitalizations in the 1st 3 months of dialysis for all consecutive patients who started dialysis between 1st January 1999 and 30th June 2002 in two Italian centers. RESULTS: The 229 patients who participated in this study (62% male, median age 70 yrs) had a high prevalence of diabetes (27.5%) and cardiovascular disease (60%). Of these, 22.7% received unstructured follow-up and 40.6% received PEP care. Peritoneal dialysis (PD) was the first choice modality for 41.5% of patients and 55% had a planned start. Excluding late referrals (36.7%), the likelihood of PD selection was similar between PEP and unstructured care (52.7% vs. 40.4%, p = NS), while that of a planned start was greater among PEP patients (91.4% vs. 38.5%, p < 0.001). Multivariate analysis showed that only PEP (but not unstructured care) vs. late referral had a significant association with reduced odds of hemodialysis (HD) selection (OR: 0.301, [95%CI 0.153, 0.594]), emergency start (0.031 [0.013, 0.075]), 1st hospitalization > 5 days (0.165 [0.081, 0.333]), and further hospitalizations (0.346 [0.174, 0.685]). Late referrals independently predicted the lack of modality selection (1.847 [1.007, 3.388]). CONCLUSIONS: A structured PEP seemed to improve early dialysis outcomes and resource usage beyond that gained by a timely referral, except for the opportunity to select modality. PMID- 14736015 TI - Multidisciplinary chronic kidney disease management improves survival on dialysis. AB - BACKGROUND: Delayed nephrology referral is associated with increased morbidity and mortality after patients begin dialysis. However, whether a pre-dialysis educational program (PEP) confers any survival advantage in comparison to unstructured specialist care is not established. METHODS: Cox's regression analysis was used to estimate the association between the type of pre-dialysis follow-up and mortality in all consecutive end-stage renal disease (ESRD) adults starting dialysis therapy in two centers, between 1 January 1999 and 30 June 2002, and followed until 30 June 2003. RESULTS: 229 patients participated in the study. The patients tended to be male (62%), elderly (median age 70 yrs) and to have cardiovascular diseases (60%). Median follow-up on dialysis was 37.8 months, with 624 patient-yrs at risk and a 0.15 yr(-1) mortality rate. Patients receiving regular unstructured care (22.7%) appeared to have similar risk for death as late referrals (36.7%), while PEP patients (40.6%) showed longer survival (hazard ratio 0.48 (95% CI 0.27, 0.87)), allowing for demographics, comorbidities, duration of the pre-dialysis follow-up and dialysis modality. Planned dialysis commencement, a better metabolic status at dialysis initiation and the previous use of angiotensin converting enzyme inhibitors were the main factors associated with improved outcomes. The survival advantage associated with PEP was still present after late referrals or observation for < or = 1 yr were excluded, as well as when survival analysis was limited to the 1st year after dialysis initiation. CONCLUSION: A multidisciplinary approach to chronic kidney diseases seems more effective than just timely referral to nephrologists in improving patient survival on dialysis. PMID- 14736016 TI - Improved health-related quality of life and left ventricular hypertrophy among dialysis patients treated with parathyroidectomy. AB - BACKGROUND: We prospectively studied changes in the perception of health-related quality of life, pruritus, and degree of left ventricular hypertrophy in end stage renal disease patients with tertiary hyperparathyroid disorder, before and 6 months after total parathyroidectomy treatment. METHODS: A series of 12 consecutive patients were enrolled. Throughout the follow-up period, all subjects completed the Kidney Disease Quality of Life Short Form-36 (KDQOL SF-36) questionnaire at inclusion and after 6 months. Serial clinical and physiological parameters including uremic pruritus, blood pressure control and left ventricular hypertrophy measured by echocardiography were recorded. RESULTS: Mean scores of the KDQOL SF-36 questionnaires were substantially higher 6 months after parathyroidectomy, with reference to physical functioning, bodily pain, role physical, role-emotional, symptom list and burden of kidney disease. Parathyroidectomy resulted in a 22% reduction in left ventricular mass index, with significant improvement from the baseline value of 246 +/- 131 to 192 +/- 131 g/m2 (p = 0.03). CONCLUSION: Our findings highlight the potential importance of parathyroidectomy in improving health-related quality of life and left ventricular hypertrophy among dialysis patients with tertiary hyperparathyroidism. PMID- 14736017 TI - Psychosocial factors and quality of life in young hemodialysis patients with low comorbidity. AB - BACKGROUND: The current predominance of older patients, diabetic patients and high-comorbidity patients among the hemodialysis (HD) population has probably influenced the definition of the effects of renal disease on health-related quality of life (HRQOL), and these effects can be different in the patient subgroup without these characteristics. This multicenter study aimed to assess HRQOL in non-diabetic HD patients, aged < 65 yrs and with low comorbidity, and to study the effects of the demographic, clinical and psychosocial characteristics on their HRQOL. METHODS: 117 patients from 43 Spanish HD centers participated in the study. Patients completed the Kidney Disease Quality of Life Short-Form questionnaire (KDQOL-SF) and screening for depressive symptoms, anxiety symptoms and social support. Various sociodemographic and clinical variables were also recorded. RESULTS: HD patients' HRQOL showed a profile similar to that of the general HD population, with low physical health scores, but normal mental health scores. Multivariate analysis demonstrated that gender, older age, non-working status, low social support and low levels of hemoglobin (Hb), Kt/V or protein catabolic rate (PCR), had a negative effects, but these effects were of relatively small magnitude and appeared only in some scales. The most important independent predictors of HRQOL were anxiety state and depressive symptoms. CONCLUSIONS: In non-diabetic HD patients, aged < or = 65 yrs and with low comorbidity, psychological factors (anxiety state and depressive symptoms) are crucial HRQOL determinants. These variables should be considered when assessing HRQOL in HD patients with these demographic and clinical characteristics. PMID- 14736018 TI - Correlation between quality of life assessment and a personality neurobiologic model in dialyzed patients. AB - The Short Form 36 Health Survey (SF-36) is a self-administered scoring system that has been widely used and validated as a quality of life (QOL) assessment tool. In our study, a cluster analysis of SF-36 scores was performed in 50 healthy volunteers (controls) and 50 neurobiologically asymptomatic patients on maintenance hemodialysis (MHD). Firstly, we assayed the tendency to form clusters from each of the investigated dimensions. Statistic analysis was performed using the Student's t-test for independent measurements and multiple regression analysis. Secondly, we attempted to evaluate if the MHD to apply to both groups a general psychobiological personality model developed by Cloninger in 1987. Cloninger describes three independent personality dimensions: novelty seeking (NS), harm avoidance (HA)and reward dependence (RD). Each personality dimension would be the expression of hereditary variations integrating the three main brain systems, respectively: dopaminergic, serotoninergic and noradrenergic. Finally, we then aimed to investigate possible interferences among the seric concentrations of the neuromodulators and SF-36 scores, in the attempt to identify, using a simple approach, the complex personality structure of MHD patients. QOL self-assessment and seric neuromodulators were measured in both groups, choosing an interdialytic day for MHD patients. We found that MHD patients perceived a significant worsening in their QOL in all investigated dimensions with respect to the controls. In addition, they showed significantly lower dopamine and serotonine concentrations and significantly higher noradrenaline concentrations. Therefore, our study, confirmed data reported previously in the literature, that cluster analysis of SF-36 scores provides different results in the MHD population in comparison to normal subjects. In fact, comparing the hierarchical trees of both groups, it appeared evident that in MHD patients, cluster dimensions were greater than in the controls. In cluster compositions showed differences between the two groups. In fact, in MHD patients there were only a few of the clusters that were observed in the controls (mental health and social functioning, vitality and general health), while role-physical and role-emotional dimensions aligned outside the hierarchical tree, with a considerable linkage distance. In our opinion, it is fascinating that the three Cloninger neuromodulators could suggest that HD patient personalities are potentially cyclothymiac, altering the disposition of the two role functions inside the hierarchical tree. PMID- 14736019 TI - Dialysis access-associated steal syndrome: the role of ultrasonography. AB - BACKGROUND: Ischemic steal syndrome is an infrequent, but potentially disabling complication of hemodialysis (HD) access creation. We analyzed the ability of duplex Doppler ultrasonography to reveal potential causes of ischemic steal syndrome in antebrachial accesses. METHODS: We performed 212 examinations on 121 patients. Ten patients suffered from ischemic steal syndrome. Complete length access evaluation was performed by a linear array 7.5 MHz ultrasound probe. RESULTS: Hand ischemia was explained by inflow artery stenosis in five cases. Excessive fistula flow due to large arteriovenous anastomosis was the suspected cause in two cases. The remaining two cases were characterized by high-resistant minimal flow in the ulnar artery with bi-directional flow in the distal part of the radial artery, suggesting stenoses located in the arcus palmaris. Duplex Doppler ultrasound had not revealed the etiology of clinically apparent hand ischemia in one case. Isolated inflow artery stenoses were treated successfully by percutaneous transluminal angioplasty. Patients with ischemic steal syndrome and high fistula flow were treated successfully by outflow vein banding. CONCLUSIONS: Duplex Doppler ultrasonography is a valuable tool for diagnosing the cause of ischemic steal syndrome and can probably replace angiography in some cases. PMID- 14736020 TI - A comparison of methods for the measurement of hemodialysis access recirculation. AB - BACKGROUND: NKF K-DOQI clinical practice guidelines recommend a regular monitoring of vascular access (VA) through several methods, such as VA recirculation (AR). Assessment of AR is crucial to avoid inefficiency of hemodialysis (HD). A non-urea based method has been developed (ultrasound dilution Transonics Hemodialysis Monitor--USM) which is considered the gold standard together with the two needle revised slow-stop-flow BUN recirculation method (S/SF). Furthermore, some other indicator dilution techniques, utilizing the dilution of serum potassium (K), glucose, hematocrit (Hct) and hemoglobin (Hb) have been recently described. METHODS: Aim of the study was to compare some of these tests with the gold standards (the USM and the revised S/SF methods). One hundred-five adult HD patients were selected to participate; all had autogenous radiocephalic wrist arteriovenous fistulas (AVF). Studies included the assessment of AR by means of: A. non-urea based methods: 1. at the start of HD a blood sample was obtained from the arterial needle at the time of needle insertion for the measurement of serum K, Hb and Hct (respectively K1, Hb1 and Hct1). The blood circuit was connected and the pumping of blood was started at 200 mL/min. After 18 seconds, blood samples were drawn from the arterial line sampling port (K2, Hb2, Hct2). At this time, if AR is present, part of the saline entering the blood stream will dilute K2, Hb2, Hct2.AR (%) is =100 x (1 - Hb2/Hb1) in the case of Hb; 2. during the first 30 min USM was performed in triplicate (the blood pump rate was set to 300 mL/min); B. a urea based method: the revised S/SF method was performed at 30 min. RESULTS: AR was absent when measured by means of the USM; it was absent even when cannulating the patients with the two needles placed as close as possible. AR measured by means of the USM was significantly different from AR measured by means of the other methods (means +/- SD, one-way analysis of variance, p < 0.0001). CONCLUSIONS: AR in autogenous radiocephalic wrist AVFs was zero when measured by means of the USM; the revised S/SF method and the tests employing the dilution of Hb, Hct and serum K can be considered a valuable alternative to the USM. PMID- 14736021 TI - Low molecular weight heparins in renal failure. AB - Low molecular weight heparins are now commonly used for systemic anticoagulation. Although elimination is mainly by the renal route, these drugs are being prescribed to patients who are dialysis dependent or have renal failure. We report 3 cases where the use of these drugs in patients with severe renal failure significantly prolonged anti-Xa activity and serious bleeding complications occurred. Concomitant prescription of other anti-thrombotic drugs may have contributed to the bleeding in 2 cases. These cases emphasize the potential toxicity of low molecular weight heparins in patients with severe renal failure and that these drugs should be used with great caution in such patients. If prescribed, doses should be lowered and anti-Xa activity frequently measured. The use of other anti-thrombotic drugs should be minimized. PMID- 14736022 TI - Unusual cases of chronic intoxication by vitamin D. AB - A 62-year-old man was hospitalized for recent renal colic and neurologic disorders. Routine biochemistry indicated the presence of hypercalcemia (serum total calcium = 15.3 mg/100 mL) and renal failure (serum creatinine = 3.72 mg/100 mL). The patient reported that he had been on treatment with a slow-release multivitamin preparation containing vitamin D and vitamin A, administered by i.m. injection. Plasma 25-OH vitamin D was > 150 ng/mL (normal range 16-74 ng/mL), plasma 1,25-(OH)2 vitamin D was 32.5 pg/mL (normal range 14-60 pg/mL), plasma parathyroid hormone 1.3 pg/mL (normal range 10-65 pg/mL). There were calcifications of left and right iliac artery at abdomen x-ray. Ultrasound and computed tomography of the glutei showed alterations of skeletal muscle and calcifications. Immediate treatment with infusion of isotonic saline, furosemide and prednisone induced rapid control of hypercalcemia and renal failure. Chronic treatment per os was discontinued after six months. The patient reported that the treatment with vitamin D had been prescribed by a physician also to his wife (55 year-old). For the woman, routine biochemistry indicated the presence of hypercalcemia (serum total calcium = 11.3 mg/100 mL) and renal failure (serum creatinine = 1.8 mg/ mL). Plasma 25-OH vitamin D was > 150 ng/mL, plasma 1,25 (OH)2 vitamin D 47.9 pg/mL, plasma parathyroid hormone was 2.5 pg/mL. Hypercalcemia was acutely treated by oral hydration, furosemide, and prednisone. Chronic treatment per os was discontinued after five months. PMID- 14736023 TI - Giovanni Maria Lancisi and urology in Rome in early modern age. AB - The "Tractatus de urinis" by Giovanni Maria Lancisi (1654-1720), dated 1696, is held by the Biblioteca Lancisiana, (the Lancisi Library, founded in 1714). It is a short manuscript devoted to the discussion of the physiological mechanisms of urine formation and excretion. It has never before been studied or published. The manuscript was probably written to be used as a didactic text for a university course at the University of Rome "La Sapienza". The analysis of Lancisi's text confirms the importance of the "iatrochemical" pattern of Italian medicine, which used the fermentation pattern to move away from a too strictly anatomical mechanistic theory, and suggests a response to the debate on the existence of an experimental practice in Rome during the Early Modern Age. PMID- 14736024 TI - Imaging kidneys: unknown documents in the history of nephrology. Five consulti by Marcello Malpighi. AB - Consulti are one of the most interesting sources for reconstructing a true outline of the state of the medical art in the Early Modern period. They were epistulae--in Latin as well as in vernacular--sent to famous physicians to receive a more correct diagnosis, and better therapeutical prescriptions. They collect and illustrate case studies, therapeutical uses, as well as the patient's attitude towards disease. This article concerns five urological consulti by Marcello Malpighi. PMID- 14736025 TI - The cellular history of the glomerulus. AB - Knowledge about the structure and functions of the cells of the glomeruli has accumulated slowly over the past 350 years. Marcello Malpighi originated the work, but it failed to progress far until Schleiden and Schwann developed their cellular theory in 1839. William Bowman linked the glomeruli to the tubules, described the parietal epithelial cells, the basement membranes, and (with Robert Todd) apparently first identified endothelial cells. Electron microscopy contributed especially to an understanding of epithelial and endothelial cell structure. Axel Key first described mesangial cells, but acceptance of these fell into abeyance for many years until Yamada incontrovertibly demonstrated their existence. Techniques such as tissue culture and molecular biological investigations have, more recently, provided much information about glomerular cell function. Progress has, throughout, depended upon the discovery of ever more powerful methods of microscopy, the development of ancillary experimental methods, the formulation of persuasive explanations for observations, and the suggestion of succinct terminology to describe the features observed. PMID- 14736027 TI - Theories and controversies on urine formation. AB - The theories of urine formation developed in the wake of progressing scientific knowledge in renal anatomy and physiology. From the philosophical theories which for a long time swung between vitalism and mechanism, the "scientific revolution" gave a great impulse to morpho/functional unit of kidney. Bowman's secretory hypothesis, as an expression of the vitalistic based theory, describes for the first time many features of the nephron and its blood supply. New insight into the inevitable errors of Bowman led Ludwig to develop the filtration-reabsorption theory, which based its scientific approach on the emerging physics and chemistry theories. The Heidenhain's secretory hypothesis which does not admit the physical filtration in Ludwig's sense, nor the hydrostatic pressure of the blood, even though incomplete and in some part without unequivocal experimental evidence, adds a fragment to the right theory of the urine formation and heralds the modern approach to the renal function of the 20th century. PMID- 14736026 TI - Double urine circulation: importance of pores. AB - The authors examine a presentation to the Royal Academy of Sciences of Paris by L. Morin, French physician and meteorologist. In this communication the presence of "pores" in the stomach and the bladder, which would allow a quick elimination of the urines on the occasion of an abundant fluid intake. PMID- 14736028 TI - Norbert Goormaghtigh and his contribution to the histophysiology of the kidney. AB - Norbert Goormaghtigh was born in Ostend on February 14, 1890 and died in Sint Martens-Latem (Belgium) on January 2, 1960. He was chief of pathology in Ghent from 1923 and became well known for his studies on the structure of the adrenal gland. He started his studies on the kidney and its juxtaglomerular apparatus at the beginning of the thirties carrying on from previous observations made by Ruiter, Borst and Zimmerman, and in 1932 he provided evidence of its endocrine function. Seminal confirming papers appeared in 1939, 1940. PMID- 14736029 TI - Investigation laws and practices in child protective services. AB - This study was triggered by the experience of one state agency when a state audit found that its investigation response and completion rates of child abuse and neglect reports did not reach 100%. At compliance rates of 99.6% and 97.58%, respectively, the auditors and news media reported a lack of compliance by the state child welfare agency. This article reviews the approaches legislatures and agencies have used to address and resolve problems of ensuring agency responsiveness without setting standards and expectations that are impossible to meet. PMID- 14736030 TI - Attitude, interest, and motivation for adoption and foster care. AB - This survey compares prospective foster and adoptive parents' attitudes, willingness, and motivations, and discusses implications for media campaigns. The results show that demographic profiles of targets for adoption and foster placements are the same, an opportunity exists to shape positive attitudes toward foster care in immigrant populations, the most compelling way to attract parents is to focus on the child in need, and testimonials of personal experiences of foster and adoptive parents should address perceived barriers to adopting or fostering. Political, religious, and environmental ideology were unrelated to attitudes or willingness to adopt or foster. Respondents with strong identifications with gay or lesbian lifestyles exhibited a higher than average willingness to adopt or foster. PMID- 14736031 TI - Collecting data on the abuse and neglect of American Indian children. AB - A survey of 10% of federally recognized American Indian tribes and the states in which they are located indicates national data systems receive reports of approximately 61% of data on the abuse or neglect of American Indian children, 42% by states and 19% by counties. The author recommends that American Indians develop culturally sound definitions of abuse and neglect and that the government provide the resources and assistance necessary to develop data tracking and reporting systems on the abuse and neglect of American Indian children. PMID- 14736032 TI - Transition from care: status and outcomes of youth who age out of foster care. AB - This study shows that a significant portion of youth exiting the foster care system face serious difficulty transitioning to life on their own. Many live on the streets, lack the money to meet basic living expenses, fail to maintain regular employment, are involved with the criminal justice system, are unable to obtain health care, and experience early pregnancies. Although youth reported exposure to independent living training while in care, few reported concrete assistance. Multiple placements while in care and less education correlated with more difficult postdischarge functioning. Training, services, positive supportive networks, and job experience in care are associated with more positive adjustments. The article advances implications for program and policy interventions. PMID- 14736033 TI - Fatal error: the missing ingredient in child welfare reform: Part 1. PMID- 14736034 TI - Exam a good tool. PMID- 14736035 TI - Exam protects public. PMID- 14736036 TI - Live patients important. PMID- 14736037 TI - Supply and demand. PMID- 14736038 TI - Patient use unethical. PMID- 14736039 TI - Translated health forms help cut through language barrier. PMID- 14736040 TI - The flu and you. PMID- 14736041 TI - Prosthetic loading of the osseointegrated implant. PMID- 14736042 TI - The role of complete denture principles in implant prosthodontics. AB - During the past 30 years, there has been a growth of implant courses in dental schools at the predoctoral level to meet a burgeoning demand. At the same time, there has been a concomitant reduction in curriculum time devoted to clinical and technical aspects of complete denture construction. There will be a sizable cohort of patients with limited natural dental landmarks seeking implant reconstructions that may be difficult for the general practitioner to restore. This article reviews basic prosthodontic principles and procedures that are important to understand when designing an implant prosthesis. PMID- 14736043 TI - When is an implant ready for a tooth? AB - The capability of placing an osseointegrated implant at the time of tooth extraction and immediately placing a restoration on the implant depends upon a number of factors. This paper describes the traditional Branemark protocol, the evolution of single-stage surgery, the guidelines for immediate placement, the measurement of implant stability, and the considerations critical to immediately loading. PMID- 14736044 TI - Immediate-loading dental endosteal implants and the elderly patient. AB - The concept of immediately loading dental implants has the potential of being a viable addition to treatment modalities. The major decision-making challenge in managing depleted dentitions and complete edentulism in an aging society lies in differentiating the treatment outcomes, especially patient-mediated assessments (including economic analyses) of the various prosthodontic options available for older adults. The ability to chew properly is of great importance to maintain a healthy nutrition and improve oral comfort and quality of life, particularly in the elderly years. PMID- 14736046 TI - Privacy act leads to paper sacks. PMID- 14736045 TI - The use of a custom template for immediate loading with the definitive prosthesis: a clinical report. AB - A new approach to implant dentistry for the fully edentulous arch is presented. Using a conventional computer tomography scan and specialized surgical planning software, a fixed final prosthesis is fabricated before the surgical procedure so the prosthesis may be delivered immediately after the implants are placed. Initial prospective studies of 10 patients with observation times of up to 24 months have shown all patients to have a functional prostheses. Long-term studies are being performed to validate this new procedure. PMID- 14736047 TI - The cat debate. PMID- 14736048 TI - A long road: Army veterinarians provide humanitarian aid, rebuild infrastructure in Iraq and Afghanistan. PMID- 14736049 TI - Emerging issues regarding informed consent. PMID- 14736051 TI - Challenges immense for infectious disease research in large animals. PMID- 14736052 TI - Parasite problem growing in goats. PMID- 14736053 TI - AAEP tackles tough issues in the Big Easy. PMID- 14736055 TI - Wishes to clarify subject of canine multiple myeloma. PMID- 14736056 TI - More confusion on precautionary principle. PMID- 14736057 TI - Additional comments on vaccination protocols. PMID- 14736058 TI - Applauds article on anesthesia myths, misconceptions. PMID- 14736059 TI - Containing cost of ACTH-stimulation test. PMID- 14736060 TI - Opinion on AVMA's animal welfare efforts. PMID- 14736061 TI - A perspective on vaccine guidelines and titer tests for dogs. PMID- 14736062 TI - What is your diagnosis? Lymphoma of high-grade malignancy. PMID- 14736063 TI - Images in veterinary dental practice. Ulcerative lymphoplasmacytic stomatitis syndrome. PMID- 14736064 TI - Anesthesia case of the month. Tachycardia. PMID- 14736065 TI - Compendium of Animal Rabies Prevention and Control, 2004. PMID- 14736066 TI - Effects of racing and nontraining on plasma thyroid hormone concentrations in sled dogs. AB - OBJECTIVE: To determine the effects of racing and nontraining on plasma thyroxine (T4), free thyroxine (fT4), thyroid-stimulating hormone (TSH), and thyroglobulin autoantibody (TgAA) concentrations in sled dogs and compare results with reference ranges established for dogs of other breeds. DESIGN: Cross-sectional study. ANIMALS: 122 sled dogs. PROCEDURE: Plasma thyroid hormone concentrations were measured before dogs began and after they finished or were removed from the Iditarod Trail Sled Dog Race in Alaska and approximately 3 months after the race. RESULTS: Concentrations of T4 and fT4 before the race were less than the reference range for nonsled dogs in 26% and 18% of sled dogs, respectively. Immediately after racing, 92% of sled dogs had plasma T4 concentrations less than the reference range. Three months after the race, 25% of sled dogs had plasma T4 concentrations less than the reference range. For T4, fT4, TSH, and TgAA, significant differences were not detected in samples collected before the race versus 3 months later. CONCLUSIONS AND CLINICAL RELEVANCE: Plasma T4, fT4, and TSH concentrations decreased in dogs that complete a long distance sled dog race. Many clinically normal sled dogs have plasma T4 and fT4 values that are lower than the reference range for nonsled dogs. We suggest that the reference ranges for sled dogs are 5.3 to 40.3 nmol/L and 3.0 to 24.0 pmol/L for plasmaT4 and fT4 concentrations, respectively, and 8.0 to 370 mU/L for TSH. PMID- 14736067 TI - Case-control study of blood type, breed, sex, and bacteremia in dogs with immune mediated hemolytic anemia. AB - OBJECTIVE: To determine whether blood type, breed, or sex were risk factors for immune-mediated hemolytic anemia (IMHA) in dogs and whether bacteremia was common in dogs with IMHA. DESIGN: Case-control study. ANIMALS: 33 dogs with IMHA, 1,014 dogs without IMHA for which blood type (dog erythrocyte antigens 1.1, 1.2, 3, 4, 5, and 7) was known, 15,668 dogs without IMHA for which breed was known, and 15,589 dogs without IMHA for which sex was known. PROCEDURE: Blood type, breed, and sex distribution of dogs with IMHA were compared with data for control dogs with Fisher exact tests and by calculating odds ratios (ORs). Results of bacterial culture of blood samples were documented for dogs with IMHA, when available. RESULTS: Dog erythrocyte antigen 7 was associated with a significant protective effect (OR, 0.1) in Cocker Spaniels with IMHA (n = 10), compared with control dogs. Cocker Spaniels, Bichon Frise, Miniature Pinschers, Rough-coated Collies, and Finnish Spitz had a significantly increased risk of IMHA, as did female dogs (OR, 2.1). Blood samples from 12 dogs with IMHA were submitted for bacterial culture, and none had bacteremia. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that blood type, breed, and sex may play a role in IMHA in dogs. PMID- 14736068 TI - Evaluation of surgical margins required for complete excision of cutaneous mast cell tumors in dogs. AB - OBJECTIVE: To determine whether neoplastic mast cells extended into tissue 1, 2, or 3 cm laterally or deeper than 1 fascial plane from the visible edge of cutaneous mast cell tumors (MCTs) in dogs. DESIGN: Prospective study. ANIMALS: 21 client-owned dogs with > or = 1 cutaneous MCT PROCEDURES: After preparation for surgery, each dog's skin was marked 1, 2, and 3 cm from the tumor edge at 0 degrees, 90 degrees, 180 degrees, and 270 degrees. At each 3-cm mark, deep fascia was exposed and sutured to the skin; the tumor was excised in routine fashion and fixed in formalin. Tumors were graded; margins were examined histologically for neoplastic mast cells. RESULTS: 23 cutaneous MCTs in 21 dogs were included in this study. Fifteen (65%) tumors were located on the trunk, 5 (22%) on the hind limbs, and 3 (13%) on the head and neck. There were 3 (13%) grade-I and 20 (87%) grade-II tumors. All grade-I tumors were completely excised at all margins. Seventy-five percent of the grade-II tumors were completely excised at the 1-cm margin, and 100% were completely excised at the 2-cm margin. Two grade-II MCTs located on the hind limbs of dogs were excised with a complete but close (within 1 mm) deep margin. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that a 2 cm lateral margin and a deep margin of 1 fascial plane appear to be adequate for complete excision of grade-I and -II MCTs in dogs. PMID- 14736069 TI - An outbreak of virulent systemic feline calicivirus disease. AB - OBJECTIVE: To describe clinical and epidemiologic features of an outbreak of feline calicivirus (FCV) infection caused by a unique strain of FCV and associated with a high mortality rate and systemic signs of disease, including edema of the face or limbs. DESIGN: Observational study. Animals-54 cats naturally infected with a highly virulent strain of FCV. PROCEDURE: Information was collected on outbreak history, clinical signs, and characteristics of infected and exposed cats. RESULTS: A novel strain of FCV (FCV-Kaos) was identified. Transmission occurred readily via fomites. Signs included edema and sores of the face and feet. Mortality rate was 40%, and adults were more likely than kittens to have severe disease (odds ratio, 9.56). Eleven (20%) cats had only mild or no clinical signs. Many affected cats had been vaccinated against FCV. Viral shedding was documented at least 16 weeks after clinical recovery. CONCLUSIONS AND CLINICAL RELEVANCE: Outbreaks of highly virulent FCV disease are increasingly common. Strains causing such outbreaks have been genetically distinct from one another but caused similar disease signs and were resistant to vaccination. All cats with suspicious signs (including upper respiratory tract infection) should be handled with strict hygienic precautions. Sodium hypochlorite solution should be used for disinfection following suspected contamination. All exposed cats should be isolated until negative viral status is confirmed. Chronic viral shedding is possible but may not be clinically important. This and similar outbreaks have been described as being caused by hemorrhagic fever-like caliciviruses, but hemorrhage is uncommonly reported. Virulent systemic FCV infection is suggested as an alternative description. PMID- 14736070 TI - Thermal cautery of the cornea for treatment of spontaneous chronic corneal epithelial defects in dogs and horses. AB - A thermal cautery technique was used to treat spontaneous chronic corneal epithelial defects (SCCEDs) in 9 eyes of 8 dogs and 2 eyes of 2 horses. Animals were sedated, and a topical anesthetic was applied. A handheld thermal cautery unit was then used to make multiple, small (< or = 1 mm in diameter), superficial burns throughout the affected area. The cautery unit was applied just until the slightest degree of contraction of the collagen fibrils was observed. After the stromal bed of the defect was treated, a rim of epithelium that extended approximately 1 mm around the denuded stroma was also subjected to thermal cautery. Following surgery, a contact lens was placed in dogs but not in horses, and the eye was treated with broad-spectrum antimicrobial ophthalmic solutions. Defects in all 11 eyes healed with minimal scarring; mean time to healing in dogs was 2.1 weeks (range, 2 to 3 weeks). The defect healed in 1 week in 1 horse and in 2 weeks in the other. Many therapeutic options are available for the treatment of SCCEDs. Procedures such as epithelial debridement and anterior stromal puncture have been shown to have a high rate of success. In cases for which these less invasive procedures fail, our results suggest that thermal cautery may be a reasonable alternative to previously described treatments for SCCEDs in dogs and horses prior to more invasive procedures such as superficial keratectomy. PMID- 14736071 TI - Management of comminuted fractures of the proximal phalanx in horses: 64 cases (1983-2001). AB - OBJECTIVE: To report the outcome of surgical treatment of comminuted fractures of the proximal phalanx in horses. DESIGN: Retrospective study. ANIMALS: 64 horses. PROCEDURE: Medical records and radiographs were reviewed to obtain information regarding signalment, fracture classification, and treatment. Follow-up information was obtained by telephone conversation or evaluation of production records. RESULTS: Thirty-eight horses had moderately comminuted fractures of the proximal phalanx. Two horses were euthanatized immediately. Fractures of the proximal phalanx in 36 horses were repaired with open reduction and internal fixation with a successful outcome in 33 (92%) horses. Reconstruction of the fracture was performed in most horses by use of a long curved incision, transection of the collateral ligament of the metacarpophalangeal or metatarsophalangeal joint, and open exposure of the proximal articular surface of the proximal phalanx. Twenty-six horses had severely comminuted fractures of the proximal phalanx. Six horses were euthanatized immediately. One horse was euthanatized after 9 days of treatment with a cast alone. Severely comminuted fractures of the proximal phalanx in 13 horses were treated with an external skeletal fixation device, and fractures healed in 8 of those horses. Six horses with severely comminuted fractures of the proximal phalanx were treated with transfixation pins incorporated into a fiberglass cast, and fractures healed in 4 horses. CONCLUSIONS AND CLINICAL RELEVANCE: Moderately comminuted fractures of the proximal phalanx can be successfully repaired; however, fractures that are too severe to permit accurate reconstruction of the fragments remain difficult to treat and horses have only a fair prognosis for survival. PMID- 14736072 TI - Exostoses of the caudal perimeter of the radial physis as a cause of carpal synovial sheath tenosynovitis and lameness in horses: 10 cases (1999-2003). AB - OBJECTIVE: To determine the clinical, radiographic, ultrasonographic, and arthroscopic findings associated with tenosynovitis of the carpal synovial sheath induced by exostoses that originate from the caudal surface of the physeal scar of the distal radius and determine the results of surgical removal of those exostoses in horses. DESIGN: Retrospective study. ANIMALS: 10 horses. PROCEDURE: Medical records of horses with effusion in the carpal synovial sheath and lameness evaluated from 1999 to 2003 were examined. RESULTS: All horses had a history of intermittent mild to moderate effusion of the carpal synovial sheath and lameness of 1 forelimb. Results of regional perineural and intrathecal anesthesia of the carpal synovial sheath confirmed that the lameness originated in the carpal synovial sheath. Radiography revealed exostoses originating from the caudal cortex of the distal radius at the level of the closed physis. Arthroscopy was performed for confirmation and removal of exostoses that penetrated the carpal synovial sheath and impinged on the deep digital flexor tendon. All horses returned to previous athletic activity. One horse had a recurrence of clinical signs 12 months after surgery, which resolved with medical treatment. CONCLUSIONS AND CLINICAL RELEVANCE: Tenosynovitis of the carpal synovial sheath and lameness were caused by impingement of exostoses of the caudal radius on the lining and contents of the carpal synovial sheath. Although the clinical signs and surgical treatment were similar to that caused by osteochondromas, these exostoses developed at the level of the closed physis of the distal radius and were not radiographically or histologically similar to osteochondromas. PMID- 14736073 TI - Segmental ostectomy of the second and fourth metacarpal and metatarsal bones in horses: 17 cases (1993-2002). AB - OBJECTIVE: To determine clinical findings in and outcome of horses with fractures of the second or fourth metacarpal or metatarsal bone that underwent segmental ostectomy, leaving the proximal and distal portions of the bone undisturbed. DESIGN: Retrospective case series. ANIMALS: 17 horses. PROCEDURES: Medical records were reviewed, and information on signalment, affected bone, lesion type, surgical procedure, amount of bone removed, and surgical and postsurgical complications was obtained. Follow-up information was obtained through telephone conversations with owners, trainers, and referring veterinarians. RESULTS: One horse had a fracture involving the distal third of the second metacarpal bone; 13 had fractures involving the middle third of the second metacarpal bone (n = 4), fourth metacarpal bone (5), or fourth metatarsal bone (4); and 3 had fractures involving the proximal third of the second (2) or fourth (1) metacarpal bone. Affected portions of the bones were surgically resected, leaving the proximal and distal portions undisturbed. All horses returned to previous performance levels without evidence of lameness. Cosmetic results were good to excellent. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that horses with a complicated injury of the proximal, middle, or distal portion of the second or fourth metacarpal or metatarsal bone may be successfully treated by means of segmental ostectomy of the abnormal portion of the bone. PMID- 14736074 TI - Outcome and subsequent fertility of sheep and goats undergoing cesarean section because of dystocia: 110 cases (1981-2001). AB - OBJECTIVE: To determine the outcome and subsequent fertility of sheep and goats undergoing a cesarean section because of dystocia. DESIGN: Retrospective study. ANIMALS: 85 sheep and 25 goats. PROCEDURE: Medical records were reviewed, and information was obtained on signalment, history, physical examination findings, anesthesia protocol, surgical technique, number of lambs or kids delivered, pre- and postoperative treatments, duration of hospitalization, and postoperative complications. Follow-up information was obtained through telephone conversations with owners. RESULTS: The proportion of sheep admitted to the veterinary teaching hospital during the study period that underwent a cesarean section (4.4%) was significantly higher than the proportion of goats that did (2.2%). Pygmy goats were overrepresented, compared with the hospital population. The most common reason for cesarean section was inadequate dilatation of the cervix. The most common surgical approach was via the left paralumbar fossa. Two hundred one lambs and kids were delivered, of which 116 were dead at delivery or died shortly afterward. Forty-two of the 65 dams with 1 or more dead fetuses had been in stage 2 labor for > 6 hours, and fetal death was significantly associated with a prolonged duration of dystocia. The most common complication following surgery was retained placenta (n = 49). Use of antimicrobials was associated with a lower rate of complications. All 16 dams that were rebred became pregnant and had no problems with dystocia. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that cesarean section is an effective method of resolving dystocia in sheep and goats and does not adversely affect subsequent fertility. PMID- 14736075 TI - Randomized treatment study of inosiplex versus combined inosiplex and intraventricular interferon-alpha in subacute sclerosing panencephalitis (SSPE): international multicenter study. AB - The efficacy of oral inosiplex alone (group A) versus combined treatment of inosiplex (Isoprinosine) and intraventricular interferon-alpha2b (Intron A) (group B) in patients with subacute sclerosing panencephalitis (SSPE) was compared. One hundred and twenty-one patients who met the diagnostic criteria for subacute sclerosing panencephalitis and presented at stage 2 or less were randomized into group A or B. Data were analyzable on 67 patients who met the inclusion criteria and adhered to the protocol. The inosiplex dosage was 100 mg/kg/day to a maximum of 3 g/day, taken orally in three divided doses for 6 months. Interferon-alpha2b started with 100,000 U/m2 and escalated to 1,000,000 U/m2 over 5 inpatient days and then 1,000,000 U/m2 twice a week for 6 months. Neurologic status was rated by the Neurological Disability Index, Brief Assessment Examination, and stages. Kaplan-Meier survival rates were not statistically significant between group A and group B (log-rank test chi2 = .1374, P = .7109). In longitudinal morbidity analyses, regression results were fitted to three outcome measures: the Neurological Disability Index, the Brief Assessment Examination, and stage. Group medians of the estimated regression slopes were then compared using the Wilcoxon rank-sum test. There was no statistically significant difference between the two groups on any of these three measures. Morbidity comparisons of clinical classification of outcomes (improvement, stabilization, worsening after treatment stopped, deterioration) also showed no statistically significant difference between groups. There were no statistically significant differences between the two treatment groups on any efficacy measure. However, the observed rates of satisfactory outcome (stabilization, improvement) of 34% in group A and 35% in group B were higher than the spontaneous remission rates of 5 to 10% reported in the literature, suggesting that treatment was superior to no treatment. PMID- 14736076 TI - Regulation of brain-derived neurotrophic factor (BDNF) expression following antibiotic treatment of experimental bacterial meningitis. AB - Although more and more new potent antibiotics have been used, mortality and neurologic deficits still occur frequently following bacterial meningitis in children. In this article, the expression of brain-derived neurotrophic factor messenger ribonucleic acid (RNA) and its production in the brains of rats were investigated during the course of experimental bacterial meningitis and after treatment with an antibiotic plus dexamethasone. In the brains of Streptococcus pneumoniae-inoculated rats, brain-derived neurotrophic factor (BDNF) messenger RNA was obviously up-regulated after inoculation for 24 hours (P < .01) and then declined but was still greater than that in the brains of control rats after inoculation for 5 days (P < .05). The expression of brain-derived neurotrophic factor in the brains of infected rats treated by antibiotic was dose dependent, down-regulated, and almost undetectable (P < .01) but up-regulated after treatment with an antibiotic plus dexamethasone (P < .01). However, the expression of brain-derived neurotrophic factor messenger RNA did not change in control rats treated with an antibiotic. Brain-derived neurotrophic factor protein showed similar changes, except it declined to normal levels 5 days after inoculation. Brain-derived neurotrophic factor messenger RNA and its production were observed in some infiltrating inflammatory cells in the brain of infected rats. The results of our studies support the hypothesis that brain-derived neurotrophic factor might play a neuroprotective role in brain damage during bacterial meningitis, and the expression of brain-derived neurotrophic factor messenger RNA and its production might be inhibited after treatment with antibiotics. The findings suggest that both eradicating the bacterial pathogen with antibiotics and adjuvant administering of brain-derived neurotrophic factor might be more beneficial to prevent brain damage. PMID- 14736077 TI - Neuropsychologic characteristics of children with attention-deficit hyperactivity disorder (ADHD), learning disorder, and tic disorder on the Rey-Osterreith Complex Figure. AB - This study examined the performance of Korean children diagnosed with attention deficit hyperactivity disorder (ADHD), learning disorder, ADHD with learning disorder, and tic disorder on the Rey-Osterreith Complex Figure. The clinical group consisted of 57 children between the ages of 6 and 13 years (15 with ADHD, 13 with learning disorder, 15 with ADHD + learning disorder, and 15 with tic disorder), and the control group included 20 children recruited from an elementary school. Rey-Osterreith Complex Figure productions were scored according to the Developmental Scoring System. The children in the control group showed a clear developmental trend--organization score increasing with age--that was similar to the performance of American children. No such age-related effect was found in the three clinical groups. The organization scores of the children with ADHD were lower than those of other groups, suggesting an organizational deficit in ADHD. Performance on the immediate recall condition was poorest in the tic disorder and ADHD + learning disorder groups, suggesting the presence of nonverbal memory problems in those groups. PMID- 14736078 TI - Neuropsychologic differences between bilateral dyskinetic and spastic cerebral palsy. AB - This study compared the neuropsychologic performance of 30 adolescents and adults with bilateral dyskinetic, mixed, and spastic cerebral palsy aged between 16 and 38 years. The sample was relatively homogeneous in terms of motor severity; no patients were able to walk unaided. In all subjects, we evaluated the general function of nonverbal reasoning and the following specific neuropsychologic areas: language, visual perception, memory, praxis, and frontal functions. Individuals with dyskinetic cerebral palsy had better auditory comprehension, visuospatial abilities, immediate visual memory, and working verbal memory than those with spastic cerebral palsy. Frontal function was the only cognitive function on which subjects with dyskinetic cerebral palsy had lower scores. We conclude that the neuropsychologic profiles of dyskinetic and spastic cerebral palsy are different. PMID- 14736079 TI - Class III beta-tubulin isotype: a key cytoskeletal protein at the crossroads of developmental neurobiology and tumor neuropathology. AB - The expression of the cytoskeletal protein class III beta-tubulin isotype is reviewed in the context of human central nervous system development and neoplasia. Compared to systemic organs and tissues, class III beta-tubulin is abundant in the brain, where it is prominently expressed during fetal and postnatal development. As exemplified in cerebellar neurogenesis, the distribution of class III beta-tubulin is neuron associated, exhibiting different temporospatial gradients in the neuronal progeny of the external granule layer versus the neuroepithelial germinal matrix of the velum medullare. However, transient expression of this protein is also present in the telencephalic subventricular zones comprising putative neuronal and/or glial precursor cells. This temporospatially restricted, potentially non-neuronal expression of class III beta-tubulin may have implications in the accurate identification of presumptive neurons derived from transplanted embryonic stem cells. In the adult central nervous system, the distribution of class III beta-tubulin is almost exclusively neuron specific. Altered patterns of expression are noted in brain tumors. In "embryonal"-type neuronal/neuroblastic tumors of the central nervous system, such as the medulloblastomas, class III beta-tubulin expression is associated with neuronal differentiation and decreased cell proliferation. In contrast, the expression of class III beta-tubulin in gliomas is associated with an ascending grade of histologic malignancy and with correspondingly high proliferative indices. Thus, class III beta-tubulin expression in neuronal or neuroblastic tumors is differentiation dependent, whereas in glial tumors, it is aberrant and/or represents "dedifferentiation" associated with the acquisition of glial progenitor-like phenotype(s). From a diagnostic perspective, the detection of class III beta-tubulin immunostaining in neoplastic cells should not be construed as categorical evidence of divergent neuronal differentiation in tumors, which are otherwise phenotypically glial. Because class III beta-tubulin is present in neoplastic but not in normal differentiated glial cells, the elucidation of molecular mechanisms responsible for the altered expression of this isotype may provide critical insights into the dynamics of the microtubule cytoskeleton in the growth and progression of gliomas. PMID- 14736080 TI - Odyssey of a pediatric neurologist in medical neuroscience teaching. PMID- 14736081 TI - Muscle glycogen depletion and increased oxidative phosphorylation following status epilepticus. AB - We describe complete glycogen depletion and increased respiratory chain enzyme activity in a muscle biopsy obtained prior to the demise of a patient in multiorgan failure following status epilepticus. These findings validate the theoretical basis of muscle energy turnover during status epilepticus: the increased demand for energy leads to complete depletion of glycogen reserves. The attempt to preserve adenosine triphosphate requirements results in increased activity of respiratory chain enzymes. PMID- 14736082 TI - Electroencephalogram and laterality of movement control: a clinical analysis. PMID- 14736083 TI - Cardioselective beta-blockers in patients with asthma and concomitant heart failure or history of myocardial infarction: when do benefits outweigh risks? PMID- 14736084 TI - Serum total antioxidant status in severe exacerbation of asthma: correlation with the severity of the disease. AB - Oxidative processes, mediated by oxygen free radicals are recognized to contribute significantly to the inflammatory pathology of bronchial asthma. An imbalance between oxidants and antioxidants has also been proposed in this disease. This study examines the serum total antioxidant status (TAS) in asthmatic patients with severe exacerbation of their disease and the probable correlation with clinical or laboratory findings. The TAS was measured in 20 patients (10 men and 10 women, with a mean age of 41.95 +/- 20.75 years), using a colorimetric method. On the days of admission and discharge, the forced expiratory volume in 1 sec (FEV1), the partial arterial oxygen pressure (PaO2), and severity criteria were recorded and correlated with TAS at the same time. The TAS was also measured in 10 healthy subjects (8 men and 2 women, mean age of 39 +/- 9 years). A statistically significant decrease of TAS was observed on admission day compared to that on discharge day (0.98 +/- 0.08 vs. 1.12 +/- 0.17 mmol/L, p < 0.001, respectively, paired t-test) suggesting the presence of oxidative stress during an asthma attack. The TAS on admission was also statistically significantly decreased compared to that of normal subjects (0.98 +/- 0.08 vs. 1.19 +/- 0.09 mmo/L, p < 0.001, respectively, paired t-test). A statistically significant correlation was observed between FEV1 change and TAS change, from admission to discharge day (r = 0.58, p = 0.007, Pearson correlation). Finally, a statistically significant correlation was found between FEV1 change and TAS on discharge day (r = 0.65, p = 0.002). Decreased TAS was found during an asthma attack, probably as a consequence of increased oxidative stress. The TAS change was correlated with severity criteria, such as FEV1. Therefore, it seems that measurement of TAS could be a simple and useful tool in the evaluation of an asthma attack. The supplementary administration of antioxidants in future needs further clarification. PMID- 14736085 TI - Difficult-to-control asthma and obstructive sleep apnea. AB - This study tested the hypothesis that asthma can promote obstructive sleep apnea (OSA) by looking at the prevalence of OSA among patients with difficult-to control asthma receiving long-term oral corticosteroid (CS) therapy and examined some possible etiological factors. The study design was a prospective cohort study and was conducted in the pulmonary outpatient clinic of a tertiary care center in Haifa, Israel. Twenty-two consecutive patients with severe unstable asthma, 14 on continuous and 8 on bursts of oral CS, in addition to their standard therapy for a mean of 8.9 +/- 3.3 years, underwent a night polysomnography in a sleep laboratory regardless of sleep complaints. A standard questionnaire was completed upon attending the sleep laboratory. The OSA was defined as respiratory disturbance index (RDI) of > or = 5 and typical complaints. The correlation between RDI to asthma and morphometric parameters was tested. All but one patient had OSA [95.5% prevalence], with mean RDI of 17.7 +/- 2.5. The RDI values were significantly higher in the continuous CS therapy subgroup (21.4 +/- 3.4 vs. 11.1 +/- 1.6, p < 0.05]. The study group had above normal neck circumferences and body mass index. The former increased by 12.1% +/- 3.1% % to 29.8% +/- 1% during the oral CS therapy interval but had no significant effect on RDI as a covariant. This study showed an unexpectedly high prevalence of OSA among patients with unstable asthma receiving long-term chronic or frequent burst of oral CS therapy. It may be assumed that prolonged and especially continuous oral CS therapy in asthma increases airway collapsibility. PMID- 14736086 TI - Patients' expectations of asthma treatment. AB - A multicomponent model has been developed to explain patients' unmet expectations of medical care. The model proposes that expectations are related to patients' personal experiences with illness, perceived vulnerability to disease, transmitted knowledge, and perceived severity of disease. The objective of this cross-sectional study was to determine whether this model can be applied to patients' unrealistic expectations of treatment outcomes, specifically expecting to be cured of asthma. In total, 230 patients observed in a primary care practice in New York City were interviewed in person with open-ended questions about their expectations of asthma treatment. Responses were analyzed with qualitative techniques to generate categories of expectations. Patients had a mean age of 41 +/- 11 years, 21% were white, 30% African American, 42% Latino, and 7% other groups. Major categories of expectations were generated from patients' responses and included symptom relief (expected by 52%), cure (36%), improved physical function (21%), and improved psychological well-being (15%). The category of expecting a cure was assessed with patients' responses to the following items representing components of the model: 1) resource utilization and medication requirements for asthma (representing severity of disease); 2) perceived quality of asthma care and satisfaction with care (representing past asthma experiences); 3) the Asthma Self-Efficacy Scale (representing perceived vulnerability to exacerbations); and 4) experiences of social network contacts with asthma and the Check Your Asthma IQ survey (representing transmitted knowledge). In bivariate analysis, patients who expected a cure were more likely to be Latino or Native American or Asian (p = 0.02), to have never required oral corticosteroids (p = 0.004), to be dissatisfied with the status of their asthma (p = 0.008), to know others who were limited by asthma (p = 0.03), to have worse Asthma Self-Efficacy Scale scores (p = 0.002), to have worse Check Your Asthma IQ scores (p = 0.04), and to currently be taking inhaled corticosteroids (p = 0.03). In multivariate analysis, worse asthma self-efficacy (p = 0.008), never having required oral corticosteroids (p = 0.005), and currently taking inhaled corticosteroids (p = 0.05) remained associated with expecting a cure. As a result of this study, we found that patients have multiple expectations of asthma treatment, including realistic expectations such as symptom relief and improved function, as well as unrealistic expectations, specifically to be cured of asthma. A multicomponent model of patient and disease characteristics was associated with this unrealistic expectation. These findings indicate that clinicians can intervene in diverse areas to foster realistic expectations of treatment outcomes among asthma patients. PMID- 14736087 TI - Endothelin-1 (ET-1) decreases human bronchial epithelial cell migration and proliferation: implications for airway remodeling in asthma. AB - The respiratory epithelium is a protective barrier that also functions as an interactive metabolically active component of the lung. The healing and repair of the epithelium involves initial migration of epithelial cells, and subsequent proliferation. The purpose of our study was to assess the effect of inflammatory mediators, in particular endothelin-1 (ET-1), on bronchial epithelial cell proliferation and migration. Under the conditions studied, ET-1 slows proliferation of human bronchial epithelial cells, compared to control (p < 0.01). The presence of ET-1 results in slower migration of epithelial cells compared to control (p < 0.04). Based on these in vitro findings, ET-1 could potentially lead to inhibition of repair of the lung epithelium and enhanced remodeling. PMID- 14736088 TI - Expression of transforming growth factor beta1 in bronchial biopsies in asthma and COPD. AB - The role of transforming growth factor beta1 (TGF beta1) in airway remodeling in asthma and chronic obstructive pulmonary disease (COPD) has not been fully described. To evaluate the possible pathogenetic role of TGF beta1 in asthma and COPD, immunohistochemical expression of TGF beta1 was described in bronchial biopsies from patients with asthma and COPD compared with healthy individuals. Twelve subjects with asthma, 13 subjects with COPD, and 10 healthy individuals enrolled in the study. Bronchial biopsies were stained with hematoxylin and eosin and anti-TGF beta1 antibody. As a result, immunoreactive TGF beta1 was mainly localized in association with connective tissue in all groups. The staining intensity was not statistically different among the groups in bronchial epithelium, whereas it was significantly higher in the group of asthma in the submucosa. Because there is evidence showing a significant increase of staining intensity in the submucosa from asthmatics but not from subjects with COPD, we may conclude that TGF beta1 may play a significant role in pathogenesis of asthma but not in COPD. PMID- 14736089 TI - Budesonide inhalation suspension reduces the need for emergency intervention in pediatric asthma: a named-patient case series. AB - This report describes 15 children aged 10-35 months frequently hospitalized with uncontrolled asthma who were monitored for a mean of 26.9 months to assess the efficacy of budesonide inhalation suspension (AstraZeneca LP, Wilmington, DE) in reducing emergency interventions. Budesonide inhalation suspension was previously shown to relieve asthma symptoms, improve pulmonary function, and reduce rescue medication use in children as young as 6 months with mild to severe persistent asthma. We now show that > or = 1 year of treatment with budesonide inhalation suspension at doses between 0.25 and 1.5 mg/d decreased the burden of asthma. The mean number of hospitalizations caused by asthma or respiratory illness decreased from 1.8 +/- 1.37 in the period before initiation of treatment with budesonide inhalation suspension to 0.33 +/- 0.62 during treatment. Likewise, the mean number of oral prednisone courses decreased from 8.1 +/- 13.7 to 1.8 +/- 2.1, and the mean number of acute respiratory illnesses requiring additional therapy decreased from 2.7 +/- 2.3 before treatment with budesonide inhalation suspension to 0.87 +/- 0.74 during treatment. Numbers of emergency department visits decreased or remained the same after initiation of budesonide inhalation suspension treatment in all but two children. There was no effect on growth rate in this group of children with moderate to severe asthma. In this case series, budesonide inhalation suspension represents a safe, effective treatment option to prevent recurrent emergency intervention for patients < 3 years of age with poorly controlled asthma. PMID- 14736090 TI - Increasing asthma awareness among physicians: impact on patient management and satisfaction. AB - Our objective was to investigate the impact of increased asthma awareness among primary care physicians on the asthma control and satisfaction of their patients. Physicians attended an asthma education session with emphasis on patient physician partnership followed by 4 month monitored follow-up of patients aged 5 44 years with mild to moderate asthma. Findings were compared with a group of patients whose physician attended the session but did not participate in the follow-up and two other control groups. The study included pediatricians and general practitioners of Maccabi Healthcare Services and their patients. Asthma symptoms were rated by patients and physicians. Data on drug prescription and use were derived from the Maccabi central database. Patient response and satisfaction and physician satisfaction were evaluated by telephone interviews. Mean asthma symptom score improved from 2.0 to 1.1 in the study group of patients (p < 0.001). The use of reliever drugs decreased concomitantly with a rise in controller drugs in all patients. An improvement in asthma status was reported by 64% of the study patients and 39% of non-participating patients (p = 0.007). Fifty-eight percent of the patients rated their competence to deal with asthma as high before the intervention compared to 62% of the participating and 55% of the non-participating patients after the intervention (p = 0.002). Most physicians claimed that simply increasing their awareness on asthma led to beneficial results in their patients. Physician education followed by monitored follow-up enhanced asthma control and patient satisfaction. Nevertheless, physician education alone appears to have a significant isolated impact on asthma control. PMID- 14736091 TI - Children's self-reports of characteristics of their asthma episodes. AB - Our purpose was to examine school-age children's self-reports of characteristics of their asthma episodes including the precipitating events, symptoms experienced during the episodes, and interventions used to resolve the episodes. Children's self-reports of their asthma episodes were assessed over a 6-week period for 42 children with persistent asthma who participated in a randomized, controlled clinical trial to evaluate the efficacy of an asthma self-management program on adherence to recommended daily peak expiratory flow rate monitoring. Children were instructed to answer the following questions on the Asthma Report Form each time they experienced an asthma episode: 1) What were you doing; 2) How did you feel; and 3) What did you do to help your breathing? Of the children, 71% experienced at least one asthma episode during the 6 weeks. There were a total of 206 episodes. Physical activity (51%) was the most cited trigger, cough alone or combined with other symptoms (84%) was the predominant symptom, and rescue asthma medication (59%) was identified most often as the intervention used by the children to resolve the asthma episode. Children's self-reports provided valuable information about their asthma episodes. The finding that most of the children experienced at least one asthma episode during the 6-week period underscores the importance of family education on how to handle asthma episodes effectively at home. Because physical activity was cited most often as a trigger for asthma episodes, families should receive education on preventive steps for averting an asthma episode prior to the child engaging in physical activity. PMID- 14736092 TI - Airway responsiveness to beta-adrenergic agonist (salbutamol) in asthma. AB - Despite the controversy of airway responsiveness to beta2-agonist drugs in asthma, in a previous study we showed increased responsiveness of asthmatic airways to isoprenaline. Therefore, in the present study of airway sensitivity to other beta2-agonists, salbutamol and its relationship to histamine responsiveness was reexamined. The threshold bronchodilator concentrations of inhaled salbutamol required for a 20% increase in forced expiratory flow in 1 sec (FEV1), (PC20) was measured in 20 normal and 19 asthmatic adults. Airway responsiveness to histamine, as the concentration that caused a 20% decrease in FEV1, was also measured in 11 normal and 12 asthmatic subjects; and the correlation between PC20 salbutamol and PC20 histamine was evaluated. Sensitivity to salbutamol was greater in asthmatics (PC20 = 7.24 mg/L) than in non-asthmatics (PC20 = 124.25 mg/L, p < 0.001). Airway responsiveness to histamine in asthmatics (PC20 = 0.18 g/L) was also significantly greater than in normal subjects (PC20 = 19.46 g/L, p < 0.001). There was a significant correlation between PC20 salbutamol and histamine (Rs = 0.6052, p < 0.005). Maximum response to both salbutamol and histamine and slope of concentration-response curves of both agents were significantly greater in patients with asthma than in normal subjects (p < 0.001 and p < 0.005 for maximum response and slope, respectively). The increased sensitivity of asthmatics to inhaled salbutamol suggests that they also may be more sensitive to their endogenous adrenaline, which may thus dilate and stabilize their airways. PMID- 14736093 TI - Primary school nurses' knowledge/competence pertaining to childhood asthma and its management prior to and following a National Asthma Education Program in Taiwan. AB - The main purpose of the National Asthma Education Program was to provide asthma education to school nurses in Taiwan. It was also designed to enhance the knowledge and competence of school nurses in managing the asthmatic problems that children experience while in school. In addition to providing instruction about current asthma management skills, tools, and other relevant information, the program demonstrated the use of the peak flow meter for asthmatic children. A single, 4-hr session conducted in each county and city in Taiwan, the National Asthma Education Program began on August 1, 1999, and ended December 31, 2000. A total of 829 school nurses joined the program, with an overall attendance rate of 74%. Significant effects of this program on nurses' asthma care knowledge and competence and case management efficacy were noted. The participating school nurses' demographics, however, were found to be irrelevant to these effects. Expecting the training activities to help relieve the anxieties of managing asthmatic cases in the school environment, participants reported that the training was of much benefit to them. Development of a teaching program to elevate school nurses' capabilities in asthmatic student care in the school environment and the implications of such a program within Taiwanese schools were also discussed. PMID- 14736094 TI - Airway remodeling of murine chronic antigen exposure model. AB - Airway remodeling is one of the most important features of bronchial asthma. However, there are few studies that have used repeated antigen exposure in murine models. We designed a murine chronic antigen exposure model necessary for studying airway remodeling. Two different strains of mice, BALB/c mice and C57BL/6 mice, were sensitized and challenged for 3-7 weeks with ovalbumin (OVA). Bronchoalveolar lavage (BAL) and histology study were conducted in each phase. Morphometry was performed, and the epithelial area ratio (Ae ratio) and subepithelial area ratio (As ratio) were calculated. The Ae ratio and As ratio of BALB/c mice were significantly increased in sensitized mice compared with non sensitized mice at 3 and 5 weeks, but not at 7 weeks. In C57BL/6 mice, the Ae ratio showed no significant changes, whereas the As ratio maintained high from 3 to 7 weeks. This thickening of the subepithelial layer consisted of collagen fibers with elastica van-Gieson (EVG) stain. Lymphocytes of the BAL showed a significant increase at 3 and 7 weeks in C57BL/6 mice, but not in BALB/c mice. A murine chronic OVA exposure model in C57BL/6 mice revealed subepithelial layer thickening consisting of collagen fibers and increased lymphocytes until 7 weeks. C57BL/6 mice are useful to elucidate the mechanism of airway remodeling. PMID- 14736095 TI - Gender differences in health-related quality of life among patients with asthma. AB - This study has a twofold objective: 1) to explore to what extent suffering from asthma affects the HRQL of men and women differently at several stages of disease severity and 2) to analyze whether the informed poorer HRQL of asthmatic women is related to their higher scores on instruments measuring emotionally disordered symptoms. One hundred fifty-one outpatient asthmatics (84 women and 67 men) completed the Spanish versions of the Asthma Quality of Life questionnaire (AQL), as well as anxiety and depression inventories. A full history, physical examination, and pulmonary function test were performed on all subjects. Patients were classified into one of four asthma severity categories following the criteria of the Global Initiative on Asthma (GINA). There were no gender differences in sociodemographic variables, asthma duration, GINA, FEV1 or dyspnea. However, women showed a poorer HRQL than men, as well as high degrees of anxiety and depression. When these data were reanalyzed taking into account the four groups of asthma severity, women only reported a poorer HRQL than men at the intermittent asthma level. The gender differences on depression and anxiety scores were maintained at the three lower severity levels, but not at the most severe asthma degree. When depression and anxiety scores were partialed out, the AQL scores maintained significant relationships with asthma severity, dyspnea, and FEV1, both in women and men. Therefore, only in men were there also relationships among AQL and sociodemographic data. The best predictor of the women's HRQL was the dyspnea score, whereas in men it was the asthma severity (GINA). PMID- 14736096 TI - Health transitions in the Pacific. PMID- 14736097 TI - Physical activity and nutritional status of adolescents on Guam. AB - The 1999 Safe & Drug Free Schools and Communities Youth Risk Behavior Study collected data from representative samples of both high school (n=590) and middle school students (n=643) on Guam in May, 1999. Results showed that the dietary and exercise habits of Guam's adolescents were sub-optimal. Out of the 643 middle school students surveyed, 26% consumed at least 3 meals per week at a fast food restaurant and 53.3% consumed at least 2 cans of sweetened soda per day. Guam adolescents had a very low intake of fruits and vegetables. As many as 75.3% of high school students reported consuming fruits and vegetables less than once a day. Students who practiced unsafe weight loss practices were significantly more likely to smoke cigarettes, and use illegal drugs. In addition, 26.9% of adolescents surveyed were considered 'overweight'--similar to the prevalence of overweight reported among U.S. Mainland adolescents. Finally, data from survey showed that adolescents on Guam spent more time watching television and less time performing various physical activities than adolescents in the U.S. mainland. These suboptimal practices put Guam's adolescents at increased risk for immediate and long-term health problems. PMID- 14736098 TI - A leptospirosis outbreak on Guam associated with an athletic event. AB - Following an "extreme-adventure" athletic event on the island of Guam, 21 of 105 participants reported being ill. Three of 10 participants who reporting having a fever (temp > 101EF) had laboratory confirmed leptospirosis. An analysis of risk factors reported by both ill and not-ill race participants suggested that exposure to recreational waters was responsible for these infections. PMID- 14736099 TI - Cholera control on Guam, 2000. AB - During April, 2000, the island of Pohnpei began experiencing an outbreak of cholera and during June and July of the same year four cases of cholera representing 3 separate introduction events were identified on Guam. Two of these events were associated with eating reef fish imported from Pohnpei. Following the imposition of a narrowly-focused ban on the importation of inshore seafood and processed food products from Pohnpei, no additional local or imported cases of cholera were detected on Guam. PMID- 14736100 TI - Estimated intake and food sources of vitamin A, folate, vitamin C, vitamin E, calcium, iron, and zinc for Guamanian children aged 9 to 12. AB - This study describes the nutrient intake and food sources of select vitamins and minerals for children on Guam. Food records (n = 954) from public school students aged nine to twelve of all regions on Guam were analyzed for nutrient content and compared to Recommended Dietary Allowances (RDA). Individual foods were condensed into 194 food aggregates and food lists representing 84% to 91% of the major vitamins and minerals in the diet of the children were developed by frequency analysis. Median intake of calcium, vitamin E, folate were less than 50% of the Recommended Dietary Allowance (RDA) and mean intake of these nutrients was 60% RDA or less. Mean and median vitamin A intake was 107% and 76% RDA, respectively. Both mean and median intake of Vitamin C, iron and zinc were present at levels above 100% RDA. Rice, meat, fruit drink from powder, milk, and fortified cereals are foods that provide substantial contributions to the vitamin and mineral content of the diets. Traditional, nutrient dense foods, such as fish, yams, papaya, and mango had minor contributions because of low frequently of consumption. Information from this study can be used to develop specific diet assessment instruments and culturally appropriate nutrition education. PMID- 14736101 TI - Missed immunisation opportunities among Tongan children in Auckland, New Zealand. AB - AIMS: To determine the proportion of Tongan children with missed opportunities for immunisations and estimate the impact of missed opportunities on delayed immunisation. METHODS: A retrospective audit of 133 children, conducted in two Pacific primary care clinics in Auckland. Children were eligible if born in 1999 and if the clinic was their usual source of primary care. Primary care computer data files up to December 31st 2001 were reviewed. A missed opportunity was defined as a doctor visit at which an immunisation was due but not given. An immunisation was defined as given only if the date of administration was recorded. An immunisation was delayed if not received within 30 days of first being due. RESULTS: Seven percent of children were delayed for the six week, 67% for the three month, 72% for the five month, and 65% for the 15 month immunisations. Thirty four percent of children had at least one missed opportunity for immunisation. Most missed opportunities occurred in acute care visits, and contraindications to immunisation were almost never present. Having a missed opportunity was associated with an increased risk of delay of the six week (OR = 48.75, 95% CI 6.82, 998.16), five month (Fisher's exact test, p = 0.03) and 15 month (Fisher's exact test, p = 0.04) immunisations but not the three month immunisation (OR = 1.00, 95% CI 0.33, 3.45). CONCLUSION: Missed opportunities are frequent and are associated with an increased risk of delayed immunisation. They are likely to contribute to New Zealand's poor immunisation coverage and the excess burden of vaccine preventable disease experienced by Pacific children in New Zealand. PMID- 14736102 TI - Smoking prevalence among young people in Papua New Guinea. AB - This study done was done during the 1996/1997-school break in NCD and Manus. There were total of 2000 and 1000 young people were interviewed respectively from NCD and Manus. From the 1150 males and 850 females interviewed in NCD, 115(10%) males and 315(37%) females were non-smokers. There were 138(12%) males and 68(8%) females smoked cigarettes. There were 150(13%) males and 84(10%) females smoked marijuana. Those who smoked marijuana also smoked cigarettes and mutrus. Even though there was higher rate of smokers among males, there is a high rate among female smokers in NCD. There were 163(10%) and 63(7%) smokers for the age group 8 10 yrs in NCD and Manus respectively. For the age group 11-12 there were 186(12%) in NCD and 91(11%) in Manus. In NCD there were 257(16%) smokers and 107(13%) in Manus in the age group of 13-14 yrs. There were 281(18%) and 153(18%) smokers in the age group 15-16 yrs in NCD and Manus respectively. There were 157(37%) of non smokers in NCD and 28(18%) in Manus did not smoke because of they were told by their teachers on the dangers of tobacco and marijuana smoking. The effort by health workers on health messages on the dangers of smoking tobacco and marijuana did influence 72(17%) non-smokers in NCD and 56(36%) in Manus on young people. Parents and teachers have full responsibility of ensuring that messages on the dangers of tobacco and marijuana smoking aregiven to the young people. Aggressive promotion of cigarette products has influenced smoking and should be stopped. PMID- 14736103 TI - A psychometric evaluation and cross-cultural validation of the 21 item Hopkins Symptom Checklist. AB - Data relating to the standardisation of a short version of the 21 item Hopkins Symptom Checklist (HSCL 21) is presented. It confirms the presence of three factors, and provides group means and percentile scales that enable comparisons to be made between the scores of a non-clinical sample and those of two separate target groups that had been under stress. These comparisons give confidence that the rating scale is appropriate for the purpose for which it was used, and suggest that the HSCL 21 could be a useful tool for clinicians in Fiji to use as an adjunct to standard interviews in the assessment of the effects of trauma. PMID- 14736104 TI - Epidemiology of infants of diabetic mothers in indigent Micronesian population Guam experience. AB - Diabetes complicating pregnancy has not yet been properly evaluated in Guam and the prevalence and morbidity of infants of diabetic mothers (IDM) in Micronesian population on Guam is described. The prevalence of IDM among the Micronesian population is 5.0% vs non-Micronesian's 3.7%. 82.5% were gestational diabetic mothers (GDM) diet controlled, 10.2% were GDM insulin controlled and 6.9% had Insulin Dependent Diabetes Mellitus. LGAs were 11% of IDMs in contrast to 6.4% of total births. Ten infants (NICU) spent total of 29 days on ventilator. Cesarean delivery, LGA, oxygen and ventilatory requirements were higher in Micronesian IDMs than in the non-Micronesian IDMs. The incidence is also higher in the Micronesian population (5.0%) compared to non Micronesian population (3.7%) on Guam. Micronesian IDMs were at higher risk for cesarean delivery, recurrent hypoglycemia, oxygen and ventilatory requirements than their non-Micronesian counterparts were. There is also a higher incidence of LGA among the Micronesian population and Chuukese had the highest incidence probably because they seek late or no prenatal care. We report 5.0% prevalence of diabetes during pregnancy in Micronesian population on Guam which imposes a significant economic burden on the local government's hospital resources. Micronesian IDMs were at higher risk for cesarean delivery, LGA, recurrent hypoglycemia, oxygen and ventilatory requirements than their non-Micronesian counterparts were. Chuukese had the highest LGA incidence in the study group. About 2/3rd of the IDM stayed 1110 extra days in hospital. IDMs accounted for the majority of expensive off-island transports. PMID- 14736105 TI - Cancer on Guam, especially among Micronesians. AB - The Guam Cancer Registry (GCR) collects data on cases of cancer in Guam residents whether their initial diagnosis is made on Guam or as a result of a referral to an off-island medical facility. It also collects data on cancer of non-residents diagnosed on Guam. Although including non-Guam resident cancer cases in statistical tables will have the effect of raising local cancer rates, this probably only partially compensates for the number of Guam-resident cancer cases that are diagnosed off-island and not reported to the registry. Cases are also collected by a periodic review of death certificates filed with the Guam Office of Vital Statistics. To test this hypothesis GCR records were reviewed to determine the number of reported cancer cases for each ethnic group that claimed to be Guam residents. Since a high proportion of cancers seen among Micronesians on Guam represent cases referred from their home islands, it may also be that these cases only represent those cancers most likely to be referred to Guam rather than being representative of the cancers most commonly occurring within these ethnic groups in their home islands. PMID- 14736106 TI - The viral diarrhoea epidemic 2002 in the Marshall Islands: its impact on a small island pharmacy. AB - This paper describes the increased demands on the pharmacy department of Majuro Hospital in the Marshall Islands during a recent viral diarrhoea epidemic. It discusses the increased number of patients, the increased usage of drugs and also the increased costs during this epidemic. The number of patients only rose 10%. However, the increase in workload of the outpatient department, laboratory and the pharmacy department was much greater. The percentage of children presenting to the outpatient department with diarrhoea increased from 10% to 80%. Oral rehydration solution (ORS) was the most common medication given during the epidemic in line with the World Health Organisation Guidelines. PMID- 14736107 TI - Guam responds to SARS. PMID- 14736108 TI - Highlights of findings from the 1999 Guam study of youth risk behaviors. PMID- 14736109 TI - Communication in health care delivery in developing countries: which way out? AB - Most governments in developing countries have adopted frameworks for health development which stressed community based initiatives and intervention at all levels of the health pyramid (WHO, 1992). But even today, most of the rural communities in these countries are still not developed in terms of available health facilities. What then is/are responsible for these failures? Various authors have come up with various reasons, principal amongst which are inadequate resources, lack of planning, insincerity/non-commitment of the governments, lack of modern information technology, etc. This paper examines some of these factors in relation to how they accentuate or hamper healthcare delivery in developing countries, using African rural communities as a study field. The resultant suggestions are a consortium of varying factors, some of which are economic in nature, policy changes, human resources development, and re-orientation of social and government attitudes towards achieving meaningful results in healthcare delivery, particularly in the rural communities. PMID- 14736110 TI - From information jungle trails to superhighways for health caregivers in the Pacific. AB - Micronesia has very sparse local library services but this is improving. For those libraries with any health science collection, only paper indexes might be available which provide limited search capabilities. Some now provide regional access to knowledge-based information in the health sciences literature with the Guam Medical Library (NHGU Library) staffed by a certified medical librarian. Over the past two decades, the authors, librarians at the NHGU Library and the RFK Library, initiated expanded and timely access to biomedical information for health professionals throughout the Pacific region. Now, health caregivers in the Pacific may affilliate for Loansome Doc service by simply contacting the NHGU Library. In 1996, with the support of the University of Guam, the RFK Library became an NLM resource library and actively began working with the Pacific Southwest Regional Medical Library Service (PSRMLS) of the NLM to provide expanded service to isolated health caregivers in Micronesia. Moreover, NHGU Library belongs to a loose network of (US) federal and Department of Defense libraries that share information and copies of articles among themselves for no cost. In this way medical services can now readily access a wider network of health information. PMID- 14736111 TI - To go or not to go organic: what choice for Pacific island countries? AB - There are arguments for and against organic agriculture. The paper looks at the impact of changes and development, the need for sustainable development, organic farming as a viable alternative, organic certification and concluding thoughts on why organic farming makes sense. To go organic does not necessarily mean that organic products are totally free of chemical pesticides but they have a lower pesticide residue. Organic certification, a process guarantee not a product guarantee, becomes necessary if a farmer has to sell organic food products locally (to tourists) or export overseas. For Pacific organic producers, the biggest obstacle lies with certification costs and initial investments in research and extension to educate consumers and farmers about organic agriculture organic methods. PMID- 14736112 TI - Introduction. Culture, change and well-being: health transitions in the Pacific. PMID- 14736113 TI - Health transition among Pacificans: unpacking imperialism. AB - What and who defines health? In the Pacific at least, health is not only an individual's state of well-being. It also refers to the positive state of the social body i.e. how people within a community interact with each other and with their environment to produce positive and desired effects. Even death can be viewed as a positive stage in human life if it facilitates the achievement of well-being as one transit or translocates to the next stage of 'eternal bliss'. The pursuit of health and well-being has resulted in many challenges over time in many communities. Health transition has been seen as a phenomenon that can be defined by the various eras of disease occurrence. Various morbidity and mortality indicators are used to measure these. According to health transition theorists, this phenomenon is linear and similar for all societies. However, this linear approach is not cognizant of the many dimensions of social, mental, physical and spiritual well-being that encompasses the life and death of Pacificans. This paper attempts to tease out the various forces that have influenced the health transition phenomenon in the Pacific. Specifically, it argues that the forces of imperialism, colonialism and globalization have largely influenced health transition in the Pacific in a manner that is to some extent, both unfavorable and oppressive. It raises the difficulty of using morbidity and mortality as measures of transition in societies where people do not die but are believed to just translocate to another life. The paper ends by suggesting alternate ways of looking at the multidimensional processes of health changes in the Pacific. PMID- 14736114 TI - Living the health transition among the Chamorros of Guam. AB - Health transition in the Pacific islands reflects the richness of diversity created by varied socio-cultural, ecological, and politico-economic systems and, therefore, requires the inclusion of the local cultural context in health assessment, planning, and program development. This paper focuses on the health transition of the Chamorros of Guam, blending assessment of health trends with the cultural contexts associated with them. The review of health trends reveals that the Chamorros are experiencing a mixed transition, including a combination of infectious and chronic diseases, in addition to the increasing impact of health risk behaviors, which have led to increasing rates of injury-related mortality. The cultural review presents Chamorro perceptions and values in their extended, multigenerational family system, in traditions of caring and sharing within their communities, and in the continued use of their traditional healers. By paying attention to the Chamorro's cultural perspectives and concerns, a "window" is provided to understand the interaction of changing population trends with their actual lives as well as an opportunity to integrate culture and health in our understanding of the health transition. PMID- 14736115 TI - Health transitions, fast and nasty: the case of Marshallese exposure to nuclear radiation. AB - The concept of health transitions assumes that health status improves with the introduction of western medicine. In this paper I demonstrate that the health of the people of Rongelap, Marshall Islands, has undergone serious damage as a result of nuclear testing, and that women in particular have suffered unduly. Exposure to nuclear radiation over a period of almost fifty years has been recognised by US authorities as a major contributory cause to the high rates of cancers and birth defects suffered by the Rongelap people. Women's reproduction has been severely affected, as evidenced by the many stillbirths and small stature of children born alive. Two generations have been exposed to both background radiation and to radiation ingested with the local foods on which they rely in the absence of other food sources. Clean up has commenced only after this and other communities sought compensation from the United States. The Rongelap people will live with the effects of radiation for generations to come. This transition to ongoing health problems is thus a negative outcome of modern health transition. PMID- 14736116 TI - The smokescreen of culture: AIDS and the indigenous in Papua, Indonesia. AB - In health transition studies on AIDS, government activities typically have been accorded less importance than local cultural practices. Certain social and cultural values, theorists argue, prevent potentially at-risk individuals from taking effective precautions to prevent HIV infection during sexual intercourse. This paper shows how a focus on culture is inadequate to understanding the issue of risk when the AIDS epidemic occurs in a colonial context. A study conducted in 2001 in Papua (West Papua), eastern Indonesia, shows that ongoing colonial relationships between indigenous Papuans and in-migrant Indonesians create inequities in AIDS awareness. Rates of HIV infection in Papua are high, but indigenous Papuans have a low level of awareness. Drawing on a survey of condom use and the sex work industry, this paper shows that government values, and economic conditions, need to be scrutinized as closely as culture in order to provide effective AIDS prevention in Papua. PMID- 14736117 TI - Reconstruction and resistance: cultural responses to living the health transition in French Polynesia. AB - This paper highlights Tahitian healing in response to rapid cultural change in French Polynesia. First, I examine the reconstruction and adaptation of Tahitian healing to cultural, economic, political, and health transitions in the past 40 years. Second, I address the issue of resistance by non-urban healers to the transformations of Tahitian healing in the urban context. Specifically, I argue that the reluctance of village healers to collaboration, association, and government legitimation (urban transitions) suggests that the status of Tahitian healing is a contested issue. The experiences of contemporary Tahitian healers challenge the unilinear health transition framework, which suggests that indigenous medicine will submit to globalization pressures and absorb a biomedical epistemology. PMID- 14736118 TI - Producing what in the transition? Health messaging and cultural constructions of health in Tonga. AB - This paper asks: what have the health promotion campaigns in Tonga actually produced, and what does this imply for our understanding of the "health transition?" Examination of villagers' responses to health messaging reveals an efflorescence of ideas about health, including a re-conceptualizing of the formal Westernized terminology of health promotion messages into concepts more aligned with traditional Tongan ways of being well. This implies the need for a model of the health transition process in which mutual shifts in understanding must be recognized by all parties, traditional villager and medicalized health professional alike, before new ideas of 'health' are actualized. PMID- 14736119 TI - 'Atamai, fakakaukau and vale: 'mind', 'thinking' and 'mental illness' in Tonga. AB - This paper explores the concepts of "mind", "thinking" and "mental illness" from a Tongan perspective. Their old Tongan equivalents are 'atamai, fakakaukau and vale. Of specific interest is mental illness. A specific state of mind, mental illness is investigated, firstly, in the conflicting relationships of mind and thinking and, secondly, in the context of the dialectical relationships between traditional forms of disease and illness and medicinal and healing concepts and practices. Their inherently clear spatio-temporal basis gives them a sense of realism. This realistic sense is much more revealing when they are positioned in the context of ta and va, Tongan for "time" and "space". Ontologically, time and space are the common medium of existence of all things, in a single level of reality. On the epistemological level, time and space are social constructs, deriving from their relative arrangement across cultures. Conflicts inevitably arising from the time-space, form-content transformation of types of human activity, create symmetries and asymmetries in the natural, mental and social realms. By integrating time and space, a general ta-va theory could be developed, so that we can better understand the complexity surrounding nature, mind and society. PMID- 14736120 TI - Diet, health and the nutrition transition: some impacts of economic and socio economic factors on food consumption patterns in the Kingdom of Tonga. AB - An essential element of the "health transition" is the emergence of disease patterns associated with changes in dietary regimes. The consumption of nutritionally poor (imported) foods in the Pacific is associated with increasing rates of diet related non-communicable diseases (NCDs). An oft-made assumption is that changes in consumption patterns are related to food preference (specifically preferences for high fat and/or dense carbohydrate foods). Recent work in the Kingdom of Tonga suggests that the "common-sense" association between food preference and food consumption is incorrect. The results of a large survey (n=430) indicate availability is the key factor in consumption, and that food preference, knowledge of the nutritional values of foods, and frequency of consumption are not correlated. Further analysis shows there are significant differences in consumption patterns between persons of higher and lower socio economic status; perception of availability and frequency of consumption are a function of economic and social position--specifically access to cash. These results underline the salience of economic factors; the rise in NCDs is correlated with the increasing importance of the cash economy (not cultural values or ignorance of nutritional issues). In the absence of economic solutions, current consumption patterns will continue. PMID- 14736121 TI - Global Public Health Conference 2002: Issues and Strategies for Hawai'i and the Pacific. PMID- 14736122 TI - Guam's quest for improved hemophilia care. PMID- 14736123 TI - Listening to the community: a first step in adapting Diabetes Today to the Pacific. AB - Diabetes is a growing problem among Pacific Islanders, but few community-based groups in the Pacific are actively working on diabetes prevention and control. The Pacific Diabetes Today Resource Center (PDTRC) was established in 1998 to adapt the Diabetes Today (DT) curriculum for Pacific Island communities in Hawai'i, American Samoa, and Micronesia. To gather data to guide the development of the Pacific Diabetes Today (PDT) curriculum, a year was spent listening to Pacific communities. First, data were gathered from health professionals on how the DT curriculum should be modified. Second, health and community leaders in 11 sites were trained and supported to conduct discussion groups with people affected by diabetes. Third, site coordinators evaluated the discussion group process. A Pacific-wide Advisory Council (AC) was established to guide the project, and the AC used findings from the first year to generate guidelines for staff to follow in adapting the DT curriculum to the Pacific. These guidelines directed staff to: a) realize that Pacific communities need to build awareness about diabetes; b) train and support local community leaders as co-facilitators in the PDT curriculum, using a learn-by-doing approach, with the goal of developing them as independent trainers; c) encourage the involvement of a broad range of community members in PDT training, including the involvement of local physicians to counter medical misconceptions about diabetes; d) give the PDT curriculum a Pacific "look" and "feel;" and e) keep the training logistically flexible to accommodate differences in communities across the region. Other programs and agencies that want to develop training programs in the Pacific may find these listening strategies and guidelines helpful. PMID- 14736124 TI - The UOG BSN program: successes and challenges. PMID- 14736125 TI - Reminiscing upon twelve and a half years of treating DM in Guam: life in the trenches. PMID- 14736126 TI - Acceleration in other axes affects +Gz tolerance: dynamic centrifuge simulation of agile flight. AB - BACKGROUND: Modern, thrust-vectored jet aircraft have the capability of developing multi-axis accelerations, especially during the performance of "supermaneuvers." These "agile" aircraft are capable of unconventional flight. The human consequences of this agile flight environment are unknown. METHODS: This multi-axis acceleration environment was studied on the Dynamic Environment Simulator gimbaled centrifuge. There were nine relaxed, unprotected subjects who were exposed to either lateral (+/- 1, +/- 2 Gy), transverse chest-to-back (+1, 2.5, or 4 Gx), or back-to-chest (-1 Gx) sustained acceleration. Positive C (+Gz) acceleration was then added beginning at 1.0 Gz by gradual onset (0.1 Gz x s(-1)) until the subjects lost nearly all of their vision. Baseline +Gz-only relaxed tolerances were measured before and after all combined Gy/Gz and Gx/Gz exposures. Heart rate, percent cerebral oxygen saturation, and cerebral blood volumes were collected during each exposure. RESULTS: Adding moderate transverse (+Gx) acceleration significantly reduced +Gz tolerance. Relaxed, unprotected +Gz tolerance was reduced approximately 0.25 G when 1.0 or 2.5 Gx was added to the increasing +Gz exposure. Adding moderate lateral Gy significantly increased +Gz tolerance. Relaxed, unprotected +Gz tolerance was increased approximately 0.5 G when +2 Gy or -2 Gy was added to the -Gz exposure. The decrease in cerebral blood volume was significantly less when +Gz was added to -1 Gx compared with the addition of +Gz to +Gx. CONCLUSIONS: Multi-axis sustained accelerations, such as those experienced during thrust-vectored aircraft maneuvers, can either enhance or reduce the +Gz tolerance of the pilot depending on the direction of the net gravitoinertial force. Gy acceleration in conjunction with Gz acceleration can enhance G tolerance. Gx acceleration in addition to Gz acceleration can reduce G tolerance. PMID- 14736127 TI - Bed rest affects ventricular and arterial elastances in monkeys: implications for humans. AB - METHODS: Experimental data were obtained from five chronically instrumented rhesus monkeys exposed to 96 h of 10 degrees head-down bed rest (HDBR) and another 96 h of 80 degrees upright control separated by 9 d of ambulatory recovery in a counter-balanced, crossover experiment design to test the hypotheses that: 1) headward and footward fluid shifts would increase systemic arterial (Eart) and left ventricular end-systolic (Ees) elastances; and 2) changes in Eart and Ees would be related in magnitude and direction. Ees and Eart were calculated from measurements taken during five observation periods for initial 2-h and 4-d exposures to HDBR that produced headward volume shifts, and acute exposure to graded levels of lower body negative pressure (LBNP) designed to produce orthostatic volume shifts. RESULTS: There was no effect of HDBR on Ees and Eart for any observation period (initial 2-h, 4-d, or LBNP). Eart increased in a similar pattern during the 4-d exposure to both control and HDBR. Ees increased with increasing LBNP levels for both control and HDBR while Eart remained unchanged. CONCLUSION: Our data are consistent with the notion that elevated Eart may represent an adaptation to physical inactivity that is associated with cardiovascular deconditioning. PMID- 14736129 TI - Neck muscle strength and endurance in fighter pilots: effects of a supervised training program. AB - INTRODUCTION: Cervical discomfort is common among pilots of high performance aircraft. An exercise program was introduced to increase the strength and endurance of the neck muscles. The purpose of this study was to analyze whether coaching or reinforcement strategies by a physical therapist was associated with improvement in neck muscle strength, endurance, and neck complaints in two cohorts of fighter pilots performing regular neck muscle exercises. METHODS: A reinforced group (RG) of 20 pilots (24-40 yr) at an Air Force base received weekly encouragement to perform their standardized exercise program three times per week. A non-reinforced reference group (NRG) of 20 pilots (23-37 yr) from another Air Force base carried out the same program without any supervision. Both groups performed the training program for 6-8 mo. Before and after the training period, isometric measurements of the neck flexors and neck extensors were performed in both groups. RESULTS: After the completion of the 6-8 mo training period, the RG pilots significantly increased their neck muscle strength (flexors: M = 3.9 nm, p = 0.000 and extensors: M = 5.0 nm, p = 0.001) as well as endurance in their neck extensors (M = 53 s, p = 0.000). The NRG pilots significantly decreased both strength (M = 11.5 nm, p = 0.0001) and endurance (M = 33 s, p = 0.003) of their neck extensors. Furthermore, the NRG pilots did not show any significant change of their neck flexor strength. No significant changes in the frequency of neck complaints were reported in either group throughout the entire study period. CONCLUSION: The reinforced training program increased the strength and endurance of the appropriate muscle groups. In order to draw any further conclusions concerning reducing neck complaints, a longer observation period with a larger group of pilots might be needed. However, it is likely that there is an individual correlation between strength and endurance of the neck muscles and neck pain, which means that any rehabilitation program should be tailored for each individual. PMID- 14736128 TI - Oxidative stress in humans during and after 4 hours of hypoxia at a simulated altitude of 5500 m. AB - BACKGROUND: High-altitude hypoxia may induce oxidative stress in humans. However, the effect of acute, severe, and non-acclimatized short-term hypobaric hypoxia exposure in humans has not been described. Additionally, little is known regarding the confounding role of reoxygenation in the extent of oxidative stress and damage markers in hypoxia. Our goals were to analyze the effect of of hypobaric hypoxia and reoxygenation on plasma oxidative stress and oxidative damage. METHODS: There were six male volunteers exposed to a simulated altitude of 5500 m (52.52 kPa) in the INEFC-UB hypobaric chamber over 4 h and returned to sea level (SL) in 30 min. Data were collected at baseline SL at 1 h and 4 h of hypoxia at 5500 m and immediately after return to sea level (RSL). RESULTS: Elevated scores of acute mountain sickness (13) and significant changes in arterial oxygen saturation (97.5 +/- 0.5; 53.3 +/- 1.9; 97.1 +/- 0.3%, p < 0.05 at SL, 4 h, and RSL, respectively) were observed. Significant reductions (p < 0.05) on total glutathione (TGSH) content were measured from SL and 1 h vs. 4 h and RSL. The percentage of oxidized glutathione (%GSSG) as an indicator of redox oxidative changes increased significantly (SL vs. 1 h; 1 h vs. 4 h, and RSL). Lipid peroxidation (TBARS), protein oxidation (SH protein groups), and total antioxidant status (TAS) followed the redox changes suggested by the glutathione system throughout the protocol. CONCLUSIONS: Hypobaric hypoxia increased the burden of plasma oxidative stress and damage markers all through the hypoxia period. However, no additional changes were observed with reoxygenation at the end of the reoxygenation period. PMID- 14736130 TI - Performance following a sudden awakening from daytime nap induced by zaleplon. AB - BACKGROUND: Zaleplon appears to be a prime candidate for assisting individuals in obtaining sleep in situations not conducive to rest (i.e., a short period during the day). However, should an early unexpected awakening and return to duty be required, the effect on performance is not known. HYPOTHESIS: Zaleplon (10 mg) would negatively affect human performance for some duration, compared with placebo, after a sudden awakening from a short period (1 h) of daytime sleep. METHODS: There were 16 participants, 8 men and 8 women, who volunteered to participate in this study. The study was conducted using a counterbalanced, double-blind, repeated measures design. At 1 h prior to drug administration, and at each of 7 h postdrug, performance measures (cognition, memory, balance, and strength) and subjective symptom reports were recorded. RESULTS: Zaleplon had a statistically significant (p < 0.05) negative impact on balance through the first 2 h postdose when compared with placebo. In addition, symptoms related to "drowsiness" were statistically more prevalent under zaleplon than under placebo through the first 3 h postdrug. Of the eight measures of cognitive performance, six were significantly negatively impacted in the zaleplon condition through 2 h postdose when compared with placebo, with one remaining significantly degraded through 3 h postdose. Zaleplon also had a significantly negative impact on memory at 1 h and 4 h postdose. CONCLUSIONS: Zaleplon (10 mg), when used as a daytime sleep aid, causes drowsiness (and related symptoms) up to 3 h postdose, and may impact task performance, especially more complex tasks, for at least 2-3 h postdose. PMID- 14736131 TI - Involuntary sleep during civil air operations: wrist activity and the prevention of sleep. AB - BACKGROUND: During air operations, aircrew may encounter sleepiness that can arise from the length of duty periods, working during the circadian low of alertness, and circadian dysrhythmia arising from time-zone change. A previous study investigated physiological and physical measures that could be used to identify sleep and provide the basis of an alertness alarm. Eye movements were sensitive indicators of sleepiness and sleep, and wrist inactivity identified sleep episodes lasting longer than 5 min. HYPOTHESIS: Wrist inactivity can provide the basis of a practical alertness device for use by aircrew. METHODS: The electrical activity of the brain (electroencephalogram, EEG), eye movements, and wrist activity were recorded continuously from the Captain and First Officer during six return flights between London and Chicago. The route comprised an outward daytime and overnight return flight, each lasting approximately 9 h. The EEG and eye movements were used to identify sleepiness and sleep, and these episodes were correlated with periods of wrist inactivity. A software simulation based on the data collected during the study was conducted in order to determine the number of errors generated by an alertness alarm based on wrist inactivity lasting 3, 4, or 5 min. RESULTS: Sleepiness or sleep was observed in 8 of the 12 subjects. All periods of sustained sleep in excess of 5 min were associated with at least 5 min of wrist inactivity. The simulated alertness alarm using a wrist inactivity setting of 5 min gave only one false alarm, and in this respect performed better than a device based on a shorter time interval (3 or 4 min). CONCLUSIONS: An alertness device based on wrist inactivity would prevent sustained periods of sleep lasting longer than 5 min. It would not, however, reliably detect sleepiness and brief episodes of sleep. The anticipated use of the device is in support of the management of alertness in aircrew, where it could be used to ensure that unintended or unauthorized napping does not occur or is detected in a timely manner. PMID- 14736132 TI - Distortion product of otoacoustic emissions as a sensitive indicator of hearing loss in pilots. AB - BACKGROUND: When the ear is presented with two simultaneous pure tones, the cochlea produces intermodulation products called distortion product otoacoustic emissions (DP-OAEs). When cochlear function is affected by noise or ototoxic drugs, DP-OAEs decrease in amplitude or disappear. The aim of this study was to investigate whether DP-OAEs are a sensitive indicator of hearing loss in pilots, for whom flying time presumably correlates with exposure to steady noise. METHODS: Subjects were 105 pilots, ages 25-44 yr, with flying times of 500-3500 hours. Pilots were divided into three groups according to their flying time and age-matched against control subjects (non-pilots) with normal hearing with no history of occupational exposure to noise. All subjects were tested by pure-tone audiometry, tympanometry, and measurement of DP-OAEs. RESULTS: DP-OAEs were significantly different among the three pilot groups, decreasing gradually at 8 frequencies with flight time. For high frequency tones (F2 7966 and 5623 Hz), age and noise were cofactors. However, 11% of the pilots in this study showed resistance to noise-induced hearing loss. DISCUSSION: DP-OAEs are a sensitive measure for monitoring early noise-induced hearing loss in pilots. Accumulating flight hours, presumed to involve steady noise exposure, initially affected high frequency hearing, then progressed to affect middle and low frequency hearing with increased flight time. There is individual variability in susceptibility and resistance to noise-induced hearing loss in humans. PMID- 14736133 TI - Human exposure to the jet fuel, JP-8. AB - INTRODUCTION: This study investigates anecdotal reports that have suggested adverse health effects associated with acute or chronic exposure to jet fuel. METHODS: JP-8 exposure during the course of the study day was estimated using breath analysis. Health effects associated with exposure were measured using a neurocognitive testing battery and liver and kidney function tests. RESULTS: Breath analysis provided an estimate of an individual's recent JP-8 exposure that had occurred via inhalation and dermal routes. All individuals studied on base exhaled aromatic and aliphatic hydrocarbons that are found in JP-8. The subject who showed evidence of the most exposure to JP-8 had a breath concentration of 11.5 mg x m(-3) for total JP-8. This breath concentration suggested that exposure to JP-8 at an Air Guard Base is much less than exposure observed at other Air Force Bases. This reduction in exposure to JP-8 is attributed to the safety practices and standard operating procedures carried out by base personnel. The base personnel who exhibited the highest exposures to JP-8 were fuel cell workers, fuel specialists and smokers, who smoked downwind from the flightline. DISCUSSION: Although study-day exposures appear to be much less than current guidelines, chronic exposure at these low levels appeared to affect neurocognitive functioning. JP-8-exposed individuals performed significantly poorer than a sample of non-exposed age- and education-matched individuals on 20 of 47 measures of information processing and other cognitive functions. PMID- 14736134 TI - Minimizing exposure to passive smoke in the enclosed environment of U.S. submarines. AB - OBJECTIVE: This study was designed to determine which designated smoking area on a submarine, forward or aft, minimizes nonsmokers' exposure to environmental tobacco smoke. METHODS: A survey was administered and urine tested by an enzyme linked immunoassay for urinary cotinine before and during a 10 d underway. There were four groups tested; submariners on boats with forward or aft designated smoking areas, with each of these groups further divided into submariners whose watch is primarily forward or aft. RESULTS: There were no significant differences in urine cotinine levels between submariners whose watches are primarily in the forward section as compared with those who work primarily aft. This was true for predeployment as compared with deployment levels and whether the designated smoking area was located forward or aft. CONCLUSION: Using cotinine as a marker, passive smoke exposure appears to be minimum. The location of the designated smoking area aboard U. S. Navy submarines does not appear to have any effect on urinary cotinine levels. PMID- 14736135 TI - Presyncopal/non-presyncopal outcomes of post spaceflight stand tests are consistent from flight to flight. AB - INTRODUCTION: The overall prevalence of orthostatic hypotension after short duration (6-18 d) spaceflight is 20% with existing countermeasures. However, it is not known if the outcomes of stand tests for orthostatic tolerance are consistent within individuals on subsequent flights, or if first time fliers are more (or less) likely to experience orthostatic hypotension and presyncope than are veteran astronauts. METHODS: There were 50 astronauts who were studied retrospectively. Stand test data, which had been collected before and after spaceflight, were compared from at least two flights for each astronaut. For 25 of these astronauts, their first flight in this database was also their first spaceflight. For the remaining 25, their first flight in this database was their second, third, or fourth flight, as data were available. RESULTS: No subject became presyncopal during preflight testing. Of the 50 subjects, 45 (90%) had the same outcome on their first and second flights of this study. Of 14 subjects on whom we had data from a third mission, 12 had the same stand test outcome on all 3 flights (86% same outcome across 3 flights). There was no correlation between flight duration and orthostatic tolerance (r = 0.39). DISCUSSION: These data support the idea that astronauts are predisposed to orthostatic tolerance/intolerance after spaceflight and that this predisposition is not altered by subsequent flights. Flight durations within this data set did not alter the likelihood of orthostatic intolerance and rookie fliers were no more likely to experience orthostatic intolerance than were veteran astronauts. PMID- 14736136 TI - High-risk head and neck movements at high G and interventions to reduce associated neck injury. AB - BACKGROUND: Neck injuries are a significant concern for aviators of high performance aircraft. A recent comprehensive technical report on cervical spinal injury associated with exposure to sustained acceleration, from NATO's Research and Technology Organization, recommended delineating the neck muscles used by aviators in this flying environment and developing improved neck muscle strengthening programs in an attempt to reduce such injuries. METHODS: A review of current literature was conducted in the fields of biomechanics, ergonomics, orthopedics, neurology, neurosurgery, rehabilitative medicine, and aerospace medicine. An objective description is provided of the muscles involved in specific head and neck movements, and those movements that are associated with a greater risk of injury during high-G sorties. The intensity and duration of force exposures common to high performance aircraft sorties, the effects of seat-back angle on these exposures, and the types and mechanisms of neck injury reported in this environment are also described. RESULTS: Primary, secondary, and tertiary preventive interventions are introduced with the goal of providing unit-level flight surgeons an approach to reducing neck injury and promoting prompt, safe return to flying of aviators with identified neck injury. A central component of these interventions is a "specific" and "intensive" neck muscle training regimen, as described in the medical literature. CONCLUSION: Increased axial compressive force and unique biomechanics combine to make neck injury likely in high performance aviators. The application of some proposed intervention strategies may reduce the occurrence of these injuries. PMID- 14736137 TI - Rupture of the pectoralis major muscle in a paratrooper. AB - Rupture of the pectoralis major muscle is a very rare injury. Excessive contraction of muscle fibers during certain forms of sports, such as weightlifting and bench pressing, is the most common cause. Among the 150 reported cases in the literature, in only 1 case did the injury happen during the landing phase of parachuting. Here we report a case of pectoralis major muscle rupture caused by a different mechanism than published previously. A paratrooper was injured during a tactical jump out of an aircraft after becoming entangled with the risers. The mechanism of injury was excessive traction and malpositioning of his shoulder when the parachute deployed. A three-phase conservative treatment regimen was performed and results were assessed by dynamometry. The patient was satisfied with the treatment and the dynamometric results were good at 9 mo after injury and at the end of a 20-mo follow-up period. We suggest that three-phase rehabilitation can be an effective treatment option for pectoralis major muscle rupture in selected patients. Prevention of this type of altitude injury would be possible by applying the fundamentals of parachuting. PMID- 14736138 TI - Emerging infectious diseases including severe acute respiratory syndrome (SARS): guidelines for commercial air travel and air medical transport. PMID- 14736139 TI - President's page. PMID- 14736140 TI - This month in aerospace medicine history--January 2004. PMID- 14736141 TI - Fly-eye collision avoidance for unmanned aerial vehicles. PMID- 14736142 TI - The history of the Aerospace Physiology Society--Part II. PMID- 14736143 TI - Advances in free-living amebae research 2003: workshop summary. PMID- 14736144 TI - Balamuthia mandrillaris: identification of clinical and environmental isolates using genus-specific PCR. PMID- 14736145 TI - Isolation of Balamuthia amebas from the environment. PMID- 14736146 TI - Genotypic identification of non-keratitis infections caused by the opportunistically pathogenic ameba genus Acanthamoeba. PMID- 14736147 TI - Occurrence and distribution of Naegleria species in thermal waters in Japan. PMID- 14736148 TI - Interaction of an Acanthamoeba human isolate harboring bacteria with murine peritoneal macrophages. PMID- 14736149 TI - Efficacy of myristamidopropyl dimethylamine (Aldox) against corneal isolates of Acanthamoeba spp. PMID- 14736150 TI - Genetic variations in the internal transcribed spacer and mitochondrial small subunit rRNA gene of Naegleria spp. AB - Naegleria spp. are widely distributed free-living amebas, but one species in the genus, N. fowleri, causes acute fulminant primary amebic meningoencephalitis in humans and other animals. Thus, it is important to differentiate N. fowleri from the rest in the genus of Naegleria, and to develop tools for the detection of intra-specific genetic variations. In this study, one isolate each of N. australiensis, N. gruberi, N. jadini, and N. lovaniensis and 22 isolates of N. fowleri were characterized at the internal transcribed spacers (ITS) and mitochondrial small subunit rRNA (mtSSU rRNA) gene. The mtSSU rRNA primers designed amplified DNA of all isolates, with distinct sequences obtained from all species examined. In contrast, the ITS primers only amplified DNA from N. lovaniensis and N. fowleri, with minor sequence differences between the two. Three genotypes of N. fowleri were found among the isolates analyzed in both the mtSSU rRNA gene and ITS. The extent of sequence variation was greater in the mtSSU rRNA gene, but the ITS had the advantage of length polymorphism. These data should be useful in the development of molecular tools for rapid species differentiation and genotyping of Naegleria spp. PMID- 14736151 TI - Workshop summary of Cryptosporidium research from the VIII International Workshop on Opportunistic Protists (IWOP-8). PMID- 14736152 TI - Microsatellite analysis of Cryptosporidium hominis and C. parvum in Portugal: a preliminary study. PMID- 14736153 TI - Cryptosporidium species and genotypes in HIV-positive patients in Lima, Peru. AB - Cryptosporidium parasites from a cross-sectional study conducted in two national hospitals in Lima, Peru were genetically characterized to determine the diversity of Cryptosporidium spp. in HIV-positive people. A total of 2,672 patients participated in this study and provided 13,937 specimens. Cryptosporidium oocysts were detected by microscopy in 354 (13.3%) of the patients. Analysis of 951 Cryptosporidium-positive specimens from 300 patients using a small subunit rRNA based PCR-RFLP tool identified 6 genotypes; Cryptosporidium hominis was the species most frequently detected (67.5%), followed by C. meleagridis (12.6%) and C. parvum (11.3%). Cryptosporidium canis (4.0%), C. felis (3.3%), and Cryptosporidium pig genotype (0.5%) were also found. These findings indicate that C. hominis is the predominant species in Peruvian HIV-positive persons, and that zoonotic Cryptosporidium spp. account for about 30% of cryptosporidiosis in these patients. PMID- 14736154 TI - Characterization of a Cryptosporidium parvum gene encoding a protein with homology to long chain fatty acid synthetase. AB - We describe here the cloning, sequencing, and characterization of a novel Cryptosporidium parvum gene, encoding a protein with significant homology to the long-chain fatty acyl-CoA synthetase (LCFA, EC 6.2.1.3). The gene has an open reading frame of 2,301 bp, coding for a 766 amino acid polypeptide, and with an estimated MW of 86.1 kDa. By indirect immunofluorescence assay, monoclonal antibodies C3CE7 and E5D labeled the anterior pole of fixed C. parvum sporozoites and developmental stages in C. parvum-infected cultures at 24, 48, and 72 h post infection. These monoclonal antibodies inhibited more than 35% of parasite growth in vitro. The effect of triacsin C, a potent selective inhibitor of LCFA synthetase, on parasite growth was assessed in cell culture; complete inhibition of parasite growth at 2.5 ug/ml was obtained with little evidence of drug associated cytotoxicity. These results suggest that the fatty acyl-CoA synthetase may be a useful target in the development of selective inhibitors and immunologic interventions against C. parvum. PMID- 14736155 TI - IL-12 knockout C57BL/6 mice are protected from re-infection with Cryptosporidium parvum after challenge. PMID- 14736156 TI - An evaluation of molecular diagnostic tools for the detection and differentiation of human-pathogenic Cryptosporidium spp. AB - The performance of 10 commonly used genotyping tools in the detection and differentiation of 7 human-pathogenic Cryptosporidium spp. (C. hominis, C. parvum, C. meleagridis, C. felis, C. canis, C. muris and Cryptosporidium pig genotype 1) was evaluated. All 3 SU rRNA gene-based tools could amplify the DNA of 7 Cryptosporidium spp. efficiently. However, the tools based on the antigens TRAP-C1, TRAP-C2 and COWP genes, the housekeeping genes HSP70 and DHFR, or a genomic sequence, failed to detect the DNA of C. felis, C. canis, Cryptosporidium pig genotype I, and C. muris. With the exception of 1 tool based on the TRAP-C2 gene, the PCR-RFLP or the PCR sequencing tools evaluated in this study could differentiate C. hominis, C. parvum and C. meleagridis from each other, and 2 SSU rRNA gene-based tools could differentiate all 7 Cryptosporidium spp. Thus, a thorough understanding of the strength and weakness of each technique is needed when using molecular diagnostic tool in epidemiological investigations of human cryptosporidiosis. PMID- 14736158 TI - Efficacy of glycoconjugated dinitroanilines against Cryptosporidium parvum. PMID- 14736157 TI - Waterborne Cryptosporidium oocyst identification and genotyping: use of GIS for ecosystem studies in Kenya and Ecuador. PMID- 14736159 TI - Molecular characterization of Cryptosporidium sp. from animals in Spain. PMID- 14736160 TI - The crystal structure of dihydrofolate reductase-thymidylate synthase from Cryptosporidium hominis reveals a novel architecture for the bifunctional enzyme. PMID- 14736161 TI - Molecular epidemiology of cryptosporidiosis in children in Malawi. AB - Few studies have examined the molecular epidemiology of cryptosporidiosis in developing countries. In this study, DNA of 69 microscopy-positive human fecal samples collected from Malawi were examined by multilocus genetic analyses. From 43, 27 and 28 of the samples, the SSU rRNA, 70 kDa heat shock protein (HSP70) and 60 kDa glycoprotein (GP60) genes, respectively, were successfully PCR-amplified. Restriction analysis of the SSU PCR products showed that 41 of the 43 PCR positive samples had C. hominis and 2 had C. parvum. Sequence analysis of the HSP70 and GP60 gene confirmed the species identification by SSU rRNA PCR-RFLP analysis, but also revealed high intraspecific variations. Altogether, six HSP70 subtypes and six GP60 subtypes (belonging to four subtype alleles) of C. hominis were found. Linkage disequilibrum analysis of the two genetic loci showed possible intraspecific recombination. Thus, cryptosporidiosis in the study area was largely caused by anthroponotic transmission. The high intraspecific variation and existence of genetic recombination were probably results of high transmission of cryptosporidiosis in this area. PMID- 14736162 TI - Loading of peptide on to MHC molecule is not essential for clearance of Cryptosporidium parvum. AB - Transgenic and knockout mice usefully model the mechanisms that result in the clearance of Cryptosporidium parvum from the gut. CD4+ cells, cells expressing MHC class II, and CD154/CD40 interactions are essential. Unexpectedly, AND RAG-/- and DO11.10 RAG-/- mice with single specificities of T cells successfully clear Cryptosporidium infection. Clearance is accompanied by activation of CD4+ cells in the MLN. The ability of T cells bearing receptors for apparently irrelevant and non-cross reactive antigens to activate and to clear infection is surprising. The requirement for class II MHC expression for Cryptosporidium clearance raises the alternative possibilities that (a) class II MHC is required to present a peptide that is loaded as a consequence of infection or (b) that the cytokine environment engendered by a Cryptosporidium infection allows affinity for self MHC to activate naive T cells. In order to test the hypothesis that peptide loading is necessary, we used A betaE alpha-/-Ii-/- mice that express a hybrid IA IE MHC molecule. They also carry a transgene that makes an E alpha peptide while disruption of their invariant chain blocks the loading of a foreign peptide on to their MHC class II molecules. After oral gavage, the course of infection was followed by ELISA. CD4+ cells in the MLN of these mice were activated to express CD69 and the infection was cleared. We conclude that the loading of a Cryptosporidium or other infection-dependent peptide onto the MHC class II molecules of APCs is not necessary for clearance of Cryptosporidium. Instead the TcR affinity for self-MHC must suffice for T cell activation in the cytokine environment resulting from infection. PMID- 14736163 TI - PCR-mediated recombination between Cryptosporidium spp. of lizards and snakes. AB - The presence or absence of genetic recombination has often been used as one of the criteria for Cryptosporidium species designation and population structure delineation. During a recent study of cryptosporidiosis in reptiles that were housed in the same room, 4 lizards were found to have concurrent infections of C. serpentis (a gastric parasite) and C. saurophilum (an intestinal parasite), and 6 snakes were concurrently infected with C. serpentis, C. saurophilum and a new Cryptosporidium as indicated by PCR-RFLP analysis of the SSU rRNA gene. DNA sequence analysis of cloned PCR products confirmed the diagnosis of mixed infections. Surprisingly, it appeared that 11 of the 22 clones (8 and 14 clones from a lizard and a snake, respectively) had chimeric sequences of two Cryptosporidium spp. BootScan analysis indicated the existence of recombinants among the cloned sequences and detection of the informative sites confirmed the BootScan results. Because the probability for genetic recombination between gastric and intestinal parasites is small, these hybrid sequences were likely results of PCR artifacts due to the presence of multiple templates. This was confirmed by PCR-sequencing analysis of single-copy templates using diluted DNA samples. Direct sequencing of 69 PCR products from 100- to 1,000-fold diluted DNAs from the same snake and lizard produced only sequences of C. serpentis, C. saurophilum and the unnamed Cryptosporidium sp. Thus, care should be taken to eliminate PCR artifacts when determining the presence of genetic recombination or interpreting results of population genetic studies. PMID- 14736164 TI - Microsporidia 2003: IWOP-8. PMID- 14736165 TI - Methionine aminopeptidase 2 expression in microsporidia. PMID- 14736166 TI - Comparison of microscopy and PCR for detection of three species of Encephalitozoon in feces. PMID- 14736167 TI - Detection of Encephalitozoon hellem in feces of experimentally infected chickens. PMID- 14736168 TI - A species of microsporidium isolated from an HIV-infected patient and identified as Trachipleistophora anthropophthera by transmission electron microscopy. PMID- 14736169 TI - Immunoblot analysis to evaluate serologic reactivity of HIV-1-negative blood donors to microsporidia. PMID- 14736170 TI - Efficiency for recovering Encephalitozoon intestinalis spores from waters by centrifugation and immunofluorescence microscopy. PMID- 14736171 TI - Microsporidia detection in stools from pets and animals from the zoo in Portugal: a preliminary study. PMID- 14736172 TI - Microsporidiosis and transplantation: a retrospective study of 23 cases. PMID- 14736173 TI - PCR-based analysis of spores isolated from smears by laser pressure catapult microdissection confirms genetic identity of spore morphotypes of the microsporidian Thelohania solenopsae. PMID- 14736174 TI - Identification of a new microsporidian parasite related to Vittaforma corneae in HIV-positive and HIV-negative patients from Portugal. AB - Fecal samples from 22 HIV-positive and 3 HIV-negative patients from Portugal with symptomatic diarrhea were diagnosed positive for microsporidia by microscopy, with most parasites detected significantly bigger than Enterocytozoon bieneusi and Encephalitozoon spp. Sequence characterization of the small subunit (SSU) rRNA gene identified a microsporidian parasite with 96% homology to two published Vittaforma corneae sequences. Phylogenetic analysis confirmed the genetic relatedness of this new microsporidian parasite to Vittaforma corneae as well as Cystosporogenes operophterae. Results of the study demonstrate the presence of a new human-pathogenic microsporidian species, which is responsible for significant number of infections in HIV-positive and HIV-negative patients in Portugal. PMID- 14736175 TI - A molecular biologic study of Enterocytozoon bieneusi in HIV-infected patients in Lima, Peru. AB - A cross-sectional study was conducted to examine the genotype distribution of Enterocytozoon bieneusi in HIV-infected patients who visited two government hospitals in Lima, Peru from January 2000 through March 2003. Microsporidia were detected by microscopy in 105 (3.9%) of 2,672 patients. A total of 212 stool samples from 89 microsporidia-positive patients were genotyped by sequence analysis of the internal transcribed spacer (ITS) region of the rRNA gene. A 392 bp fragment containing the complete ITS region was amplified and sequenced. Multiple alignments and phylogenetic analysis of these ITS sequences identified 11 distinct genotypes of E. bieneusi (Peru-1 to Peru-11), 6 of which were new genotypes not reported before. The remaining 5 genotypes had nucleotide sequences identical to those previously reported in humans, cats, pigs, and wild mammals. All the 11 E. bieneusi-genotypes identified are genetically related, and members of the group have been previously found in humans, domestic animals, and some wild mammals. Thus, there is a high genetic diversity of E. bieneusi in humans in Peru, and zoonotic transmission is possible if humans are in close contact with infected animals. PMID- 14736176 TI - Characterization of recombinant microsporidian methionine aminopeptidase type 2. PMID- 14736177 TI - Lectin binding of the major polar tube protein (PTP1) and its role in invasion. PMID- 14736178 TI - Summary of pneumocystis research presented at the 8th International Workshop on Opportunistic Protists. PMID- 14736179 TI - Expression profiling of the responses of Pneumocystis carinii to drug treatment using DNA macroarrays. PMID- 14736180 TI - Dihydropteroate synthase (DHPS) genotyping by PCR-RFLP analysis of Pneumocystis jirovecii repeated isolates from HIV-infected patients: a preliminary study. PMID- 14736181 TI - Pneumocystis dihydropteroate synthase mutations in patients with Pneumocystis pneumonia who are newly diagnosed with HIV infection. PMID- 14736182 TI - Influence of climatic factors on Pneumocystis carriage within a socially organized group of immunocompetent macaques (Macaca fascicularis). AB - As monkeys-derived Pneumocystis is closely related to P. jirovecii, simian populations should be considered as valuable models for the understanding of the epidemiology of human pneumocystosis. In the present study, the impact of environmental factors on the carriage of Pneumocystis was evaluated in socially organized group of immunocompetent macaques (Macaca fascicularis). The tribe, maintained in partial release at the Primatology Center of Strasbourg in France, comprised 29 animals at the end of the study. From December 2000 to November 2002, deep nasal swab samples were collected monthly from each animal under general anaesthesia. The presence of Pneumocystis DNA was assessed by nested PCR of mtLSU rRNA gene. No case of pneumocystosis was reported during the study. Pneumocystis DNA was detected in 166 out of 481 swab samples examined (34.5%). The number of macaques with detectable Pneumocystis DNA was highly variable from one month to another. However, Pnemocystis carriage was clearly correlated to the mean precipitation rates. PMID- 14736183 TI - Long-term colonization with Pneumocystis jirovecii in hospital staffs: a challenge to prevent nosocomial pneumocystosis. PMID- 14736184 TI - Pneumocystis colonization in HIV-infected patients. PMID- 14736185 TI - Application of an mRNA-based molecular viability assay to oropharyngeal washes for the diagnosis of Pneumocystis pneumonia in HIV-infected patients. A pilot study. PMID- 14736186 TI - Melanin-like pigments in Pneumocystis carinii. PMID- 14736187 TI - Evidence that biosynthesis of individual ubiquinone homologs in Pneumocystis carinii is under homolog-specific negative feedback (product) control. PMID- 14736188 TI - Evolution and speciation of Pneumocystis. PMID- 14736189 TI - Sequence diversity of transcripts from Pneumocystis carinii gene families MSR and PRT1. PMID- 14736190 TI - Microarray analysis of lung epithelial responses to Pneumocystis carinii. PMID- 14736191 TI - Decrease in alveolar macrophage number during Pneumocystis carinii infection. PMID- 14736192 TI - Environmental stress and encystment of Pneumocystis carinii. PMID- 14736193 TI - Inducible nitric oxide synthase expression in Pneumocystis carinii pneumonia. PMID- 14736194 TI - Life cycle stage-specific and encystment protein profiles in Pneumocystis carinii. PMID- 14736195 TI - Numbers of alveolar macrophages are increased during Pneumocystis pneumonia in mice. PMID- 14736196 TI - Pneumocystis carinii cyst is associated with inflammation in the host. PMID- 14736197 TI - Pneumocystis carinii factor responsible for reduced phagocytic activity in alveolar macrophages. PMID- 14736198 TI - Transtracheal inoculation of Pneumocystis carinii in immunocompetent animals. PMID- 14736199 TI - Transcriptional changes in alveolar macrophages during Pneumocystis pneumonia. PMID- 14736201 TI - Pneumocystis jirovecii carriage in Portuguese immunocompetent patients: preliminary results. PMID- 14736200 TI - Pneumocystis carinii: cell wall beta-glucan-mediated pulmonary inflammation. PMID- 14736202 TI - Seroprevalence of Pneumocystis human infection in southern Spain. PMID- 14736203 TI - Real time quantitative PCR assay for Pneumocystis jirovecii detection. PMID- 14736204 TI - Site-directed mutagenesis of the Saccharomyces cerevisiae dihydropteroate synthase FOL1 gene to study Pneumocystis jirovecii mutations in the orthologue gene FAS. PMID- 14736205 TI - Multilocus genotyping of Pneumocystis jirovecii from adult HIV-infected patients with Pneumocystis pneumonia. PMID- 14736206 TI - Pneumocystis jirovecii dihydrofolate reductase polymorphisms associated with failure of prophylaxis. PMID- 14736207 TI - Development of a real-time PCR-based fluorescence assay for rapid detection of point mutations in Pneumocystis jirovecii dihydropteroate synthase gene. PMID- 14736209 TI - Generation of sequencing libraries for the Pneumocystis Genome project. PMID- 14736208 TI - Immune responses to Pneumocystis colonization and infection in a simian model of AIDS. PMID- 14736210 TI - Large doses of major surface glycoprotein-sensitized donor splenocytes transferred to corticosteroid treated rats with Pneumocystis pneumonia result in successful reduction of organism burden. PMID- 14736211 TI - Immunocompetent infants as a human reservoir for Pneumocystis jirovecii: rapid screening by non-invasive sampling and real-time PCR at the mitochondrial large subunit rRNA gene. AB - We tested a real-time PCR assay targeting the Pneumocystis jirovecii mitochondrial large subunit rRNA gene on 240 archival nasopharyngeal aspirates from non-immunosuppressed infants. The sensitivity of this assay appeared close to that of a conventional nested-PCR assay targeting the same locus. Because of its one-step procedure, and its sensitivity and rapidity, the real-time PCR assay is particularly suitable for screening individuals parasitized by P. jirovecii within large populations. PMID- 14736212 TI - Multilocus genotyping of Pneumocystis jirovecii in patients developing diverse forms of parasitism: implication for a wide human reservoir for the fungus. AB - Pneumocystis jirovecii ITS and DHPS genotypes were identified in 3 patient groups developing diverse forms of P. jirovecii infections: 13 patients with Pneumocystis pneumonia, 8 patients merely colonized by the fungus, and 19 immunocompetent infants with bronchiolitis developing mild P. jirovecii infection. Common P. jirovecii genotypes were found in the 3 patient groups, suggesting that common sources of P. jirovecii were involved in the fungus acquisition, and that transmission cycles of P. jirovecii infections in these patient groups are not independent. Parasitized patients, whatever the form of parasitism they present, may be part of a common reservoir for P. jirovecii. PMID- 14736213 TI - Lymphocyte response in subjects with chronic pulmonary disease colonized by Pneumocystis jirovecii. PMID- 14736214 TI - Complementation and in vivo biochemical characterization of the Pneumocystis carinii MAPK. PMID- 14736215 TI - Pneumocystis carinii BCK1 complements the Saccharomyces cerevisiae cell wall integrity pathway. PMID- 14736216 TI - Evidence for cholesterol scavenging by Pneumocystis and potential modifications of host-synthesized sterols by the P. carinii SAM:SMT. PMID- 14736217 TI - Definitive structural identities of Pneumocystis jirovecii sterols. PMID- 14736218 TI - Isospora belli infection: observation of unicellular cysts in mesenteric lymphoid tissues of a Brazilian patient with AIDS and animal inoculation. AB - We describe the finding of unizoic cysts of Isospora belli in lymphoid tissues of a Brazilian patient with AIDS, and discuss the possibilities of their drug resistance, they being the cause of relapses, and of being an indication for the existence of intermediary or paratenic animal hosts. PMID- 14736219 TI - Cell death mechanisms in the human opportunistic pathogen Candida albicans. AB - Difficulties arising during chemotherapy of Candida albicans necessitate novel chemotherapeutic strategies. Garlic extract and two of its constituents, diallyl disulphide and allyl alcohol, are potentially useful anti-candidal agents. Flow Cytometry has been used to measure the population distributions of apoptotic/necrotic cell death using annexin V-FITC/propidium iodide and oxidative stress dichlorodihydrofluorescein. Candicidal mechanisms may be due to programmed cell death induced by oxidative stress, mediated by the generation of reactive oxygen species or alternatively by the depletion of cellular thiols, which normally act as redox buffer systems for defence. We suggest that mechanisms that these anti-candidal agents have in common is the triggering some of the characteristics of apoptotic cell death. PMID- 14736220 TI - Sporulation and survival of Toxoplasma gondii oocysts in seawater. AB - We have been collaborating since 1992 in studies on southern sea otters (Enhydra lutris nereis) as part of a program to define factors, which may be responsible for limiting the growth of the southern sea otter population. We previously demonstrated Toxoplasma gondii in sea otters. We postulated that cat feces containing oocysts could be entering the marine environment through storm run-off or through municipal sewage since cat feces are often disposed down toilets by cat owners. The present study examined the sporulation of T. gondii oocysts in seawater and the survival of sporulated oocysts in seawater. Unsporulated oocysts were placed in 15 ppt artificial seawater, 32 ppt artificial seawater or 2% sulfuric acid (positive control) at 24 C in an incubator. Samples were examined daily for 3 days and development monitored by counting 100 oocysts from each sample. From 75 to 80% of the oocysts were sporulated by 3 days post-inoculation under all treatment conditions. Groups of 2 mice were fed 10,000 oocysts each from each of the 3 treatment groups. All inoculated mice developed toxoplasmosis indicating that oocysts were capable of sporulating in seawater. Survival of sporulated oocysts was examined by placing sporulated T. gondii oocysts in 15 ppt seawater at room temperature 22-24 C (RT) or in a refrigerator kept at 4 C. Mice fed oocysts that had been stored at 4C or RT for 6 months became infected. These results indicate that T. gondii oocysts can sporulate and remain viable in seawater for several months. PMID- 14736221 TI - Mode of action of ponazuril against Toxoplasma gondii tachyzoites in cell culture. AB - Toxoplasma gondii is an important apicomplexan parasite of humans and other warm blooded animals. Ponazuril is a triazine anticoccidial recently approved for use in horses in the United States. We investigated the mode of action of ponazuril against developing RH strain T. gondii tachyzoites in African green monkey kidney cells. Host cells were infected with 2.0 x 10(5) tachyzoites and treated with 5 microg/ml ponazuril. Cultures were fixed and examined by transmission electron microscopy 3 days after treatment. Ponazuril interfered with normal parasite division. This led to the presence of multinucleate schizonts stages. Up to six tachyzoites were observed partially budded from the surface of these schizonts. Large vacuoles developed in these schizonts and they eventually degenerated. PMID- 14736222 TI - Emergence of zoonotic canine leishmaniasis in the United States: isolation and immunohistochemical detection of Leishmania infantum from foxhounds from Virginia. AB - Previously considered an exotic disease, canine leishmaniasis caused by Leishmania infantum has recently been detected within the foxhound population in the United States and parts of Canada. Leishmania infantum is the etiologic agent of visceral leishmaniasis in many areas of the world and dogs are considered a major reservoir host for human Leishmania infections. Human visceral leishmaniasis has recently emerged as an opportunistic infection among individuals co-infected with HIV/AIDS and in persons taking immunosuppressive drugs. We report the isolation of L. infantum from 3 naturally infected foxhounds from Virginia by culture of popliteal lymph node and bone marrow, and the development of an immunohistochemical test to detect the parasite in tissues. PMID- 14736223 TI - Summary: International conference on anaerobic protists. PMID- 14736224 TI - A 39-kDa cysteine proteinase CP39 from Trichomonas vaginalis, which is negatively affected by iron may be involved in trichomonal cytotoxicity. PMID- 14736225 TI - A comparison of PCR and in vitro culture for detection of Tritrichomonas foetus in bovine preputial scrapings. PMID- 14736226 TI - Effect of varying cobalt-60 doses on survival and growth of Giardia lamblia trophozoites. PMID- 14736227 TI - Two Trichomonas vaginalis loci encoding for distinct cysteine proteinases show a genomic linkage with putative inositol hexakisphosphate kinase (IP6K2) or an ABC transporter gene. PMID- 14736228 TI - Purification of Entamoeba histolytica DNA containing organelles (EkhOs): a further characterization. PMID- 14736229 TI - First report of Giardia in coyotes (Canis latrans). PMID- 14736230 TI - Genomic polymorphism among Blastocystis isolates and development of PCR-based identification of zoonotic isolates. PMID- 14736231 TI - Scaffold hopping in de novo design. Ligand generation in the absence of receptor information. AB - We report here the de novo generation of chemotypes and scaffolds for the estrogen receptor, without use of the receptor structure in the assembly phase. Through use of ligand superpositions or a single bound conformation of a known active, a pseudoreceptor can be generated as a design envelope, within which novel structures are readily assembled. Many of these structures have high similarity to known chemotypes. Scaffold hopping is readily achieved within this pseudoreceptor, indicating the advantages of such an approach in discovery research. PMID- 14736232 TI - Design, synthesis, and evaluation of metabolism-based analogues of haloperidol incapable of forming MPP+-like species. AB - The long-term, irreversible, Parkinsonism-like side effects of haloperidol have been speculated to involve several mechanisms. More recently, it has been speculated that the metabolic transformation to MPP+-like species may contribute to the Parkinsonism-like side effects. Because BCPP+ and its reduced analogue have been shown to possess the potential to destroy dopamine receptors in the nigrostriatum, we have designed new analogues of haloperidol lacking the structural features necessary to form neurotoxic quaternary species but retaining their dopamine-binding capacity. The most potent agent at the D2 receptor, the homopiperidine analogue 11, was found to be equipotent to haloperidol. It was also of interest to identify analogues with DA binding profiles similar to that of clozapine at the dopamine receptor subtypes. Evaluation of the proposed agents shows that the ratio of D2 to D4 (2) binding of clozapine was mimicked by 7 [K(i)(D2) = 33, K(i)(D3) = 200, K(i)(D4) = 11 nM; K(i)(D2)/K(i)(D4) = 3] and 9 [K(i)(D2) = 44, K(i)(D3) = 170, K(i)(D4) = 24 nM; K(i)(D2)/K(i)(D4) = 2]. A preliminary in-vivo testing of compound 7 shows that its behavioral profile is similar to that of clozapine. This profile suggests that there is a need for further evaluation of these two synthetic agents and their enantiomers for efficacy and lack of catalepsy in animal models. PMID- 14736233 TI - Inhibition of the bacterial enoyl reductase FabI by triclosan: a structure reactivity analysis of FabI inhibition by triclosan analogues. AB - To explore the molecular basis for the picomolar affinity of triclosan for FabI, the enoyl reductase enzyme from the type II fatty acid biosynthesis pathway in Escherichia coli, an SAR study has been conducted using a series of triclosan analogues. Triclosan (1) is a slow, tight-binding inhibitor of FabI, interacting specifically with the E.NAD(+) form of the enzyme with a K(1) value of 7 pM. In contrast, 2-phenoxyphenol (2) binds with equal affinity to the E.NAD(+) (K(1) = 0.5 microM) and E.NADH (K(2) = 0.4 microM) forms of the enzyme and lacks the slow binding step observed for triclosan. Thus, removal of the three triclosan chlorine atoms reduces the affinity of the inhibitor for FabI by 70,000-fold and removes the preference for the E.NAD(+) FabI complex. 5-Chloro-2-phenoxyphenol (3) is a slow, tight-binding inhibitor of FabI and binds to the E.NAD(+) form of the enzyme (K(1) = 1.1 pM) 7-fold more tightly than triclosan. Thus, while the two ring B chlorine atoms are not required for FabI inhibition, replacement of the ring A chlorine increases binding affinity by 450,000-fold. Given this remarkable observation, the SAR study was extended to the 5-fluoro-2 phenoxyphenol (4) and 5-methyl-2-phenoxyphenol (5) analogues to further explore the role of the ring A substituent. While both 4 and 5 are slow, tight-binding inhibitors, they bind substantially less tightly to FabI than triclosan. Compound 4 binds to both E.NAD(+) and E.NADH forms of the enzyme with K(1) and K(2) values of 3.2 and 240 nM, respectively, whereas compound 5 binds exclusively to the E.NADH enzyme complex with a K(2) value of 7.2 nM. Thus, the ring A substituent is absolutely required for slow, tight-binding inhibition. In addition, pK(a) measurements coupled with simple electrostatic calculations suggest that the interaction of the ring A substituent with F203 is a major factor in governing the affinity of analogues 3-5 for the FabI complex containing the oxidized form of the cofactor. PMID- 14736234 TI - Structural analysis and optimization of NK(1) receptor antagonists through modulation of atropisomer interconversion properties. AB - We have previously described a series of antagonists that showed high potency and selectivity for the NK(1) receptor. However, these compounds also had the undesirable property of existing as a mixture of interconverting rotational isomers. Here we show that alteration of the 2-naphthyl substituent can modulate the rate of isomer exchange. Comparisons of the NK(1) receptor affinity for the various conformational forms has facilitated the development of a detailed NK(1) pharmacophore model. PMID- 14736235 TI - Discovery of 4-substituted pyrrolidone butanamides as new agents with significant antiepileptic activity. AB - (S)-alpha-ethyl-2-oxopyrrolidine acetamide 2 (levetiracetam, Keppra, UCB S.A.), a structural analogue of piracetam, has recently been approved as an add-on treatment of refractory partial onset seizures in adults. This drug appears to combine significant efficacy and high tolerability due to a unique mechanism of action. The latter relates to a brain-specific binding site for 2 (LBS for levetiracetam binding site) that probably plays a major role in its antiepileptic properties. Using this novel molecular target, we initiated a drug-discovery program searching for ligands with significant affinity to LBS with the aim to characterize their therapeutic potential in epilepsy and other central nervous system diseases. We systematically investigated the various positions of the pyrrolidone acetamide scaffold. We found that (i) the carboxamide moiety on 2 is essential for affinity; (ii) among 100 different side chains, the preferred substitution alpha to the carboxamide is an ethyl group with the (S) configuration; (iii) the 2-oxopyrrolidine ring is preferred over piperidine analogues or acyclic compounds; (iv) substitution of positions 3 or 5 of the lactam ring decreases the LBS affinity; and (v) 4-substitution of the lactam ring by small hydrophobic groups improves the in vitro and in vivo potency. Six interesting candidates substituted in the 4-position have been shown to be more potent antiseizure agents in vivo than 2. Further pharmacological studies from our group led to the selection of (2S)-2-[(4R)-2-oxo-4-propylpyrrolidin-1 yl]butanamide 83alpha (ucb 34714) as the most interesting candidate. It is approximately 10 times more potent than 2 as an antiseizure agent in audiogenic seizure-prone mice. A clinical phase I program has been successfully concluded and 83alpha will commence several phase II trials during 2003. PMID- 14736236 TI - Unexpected nanomolar inhibition of carbonic anhydrase by COX-2-selective celecoxib: new pharmacological opportunities due to related binding site recognition. AB - By optimizing binding to a selected target protein, modern drug research strives to develop safe and efficacious agents for the treatment of disease. Selective drug action is intended to minimize undesirable side effects from scatter pharmacology. Celecoxib (Celebrex), valdecoxib (Bextra), and rofecoxib (Vioxx) are nonsteroidal antiinflammatory drugs (NSAIDs) due to selective inhibition of inducible cyclooxygenase COX-2 while sparing inhibition of constitutive COX-1. While rofecoxib contains a methyl sulfone constituent, celecoxib and valdecoxib possess an unsubstituted arylsulfonamide moiety. The latter group is common to many carbonic anhydrase (CA) inhibitors. Using enzyme kinetics and X-ray crystallography, we demonstrate an unexpected nanomolar affinity of the COX-2 specific arylsulfonamide-type celecoxib and valdecoxib for isoenzymes of the totally unrelated carbonic anhydrase (CA) family, such as CA I, II, IV, and IX, whereas the rofecoxib methyl sulfone-type has no effect. When administered orally to glaucomatous rabbits, celecoxib and valdecoxib lowered intraocular pressure, suggesting that these agents may have utility in the treatment of this disorder. The crystal structure of celecoxib in complex with CA II reveals part of this inhibition to be mediated via binding of the sulfonamide group to the catalytic zinc of CA II. To investigate the structural basis for cross-reactivity of these compounds between COX-2 and CA II, we compared the molecular recognition properties of both protein binding pockets in terms of local physicochemical similarities among binding site-exposed amino acids accommodating different portions of the drug molecules. Our approach Cavbase, implemented into Relibase, detects similarities between the sites, suggesting some potential to predict unexpected cross-reactivity of drugs among functionally unrelated target proteins. The observed cross-reactivity with CAs may also contribute to differences in the pharmacological profiles, in particular with respect to glaucoma and anticancer therapy and may suggest new opportunities of these COX-2 selective NSAIDs. PMID- 14736237 TI - Evaluation of docking performance: comparative data on docking algorithms. AB - Docking molecules into their respective 3D macromolecular targets is a widely used method for lead optimization. However, the best known docking algorithms often fail to position the ligand in an orientation close to the experimental binding mode. It was reported recently that consensus scoring enhances the hit rates in a virtual screening experiment. This methodology focused on the top ranked pose, with the underlying assumption that the orientation/conformation of the docked compound is the most accurate. In an effort to eliminate the scoring function bias, and assess the ability of the docking algorithms to provide solutions similar to the crystallographic modes, we investigated the most known docking programs and evaluated all of the resultant poses. We present the results of an extensive computational study in which five docking programs (FlexX, DOCK, GOLD, LigandFit, Glide) were investigated against 14 protein families (69 targets). Our findings show that some algorithms perform consistently better than others, and a correspondence between the nature of the active site and the best docking algorithm can be found. PMID- 14736238 TI - Synthesis and antiviral activity of (Z)- and (E)-2,2 [bis(hydroxymethyl)cyclopropylidene]methylpurines and -pyrimidines: second generation methylenecyclopropane analogues of nucleosides. AB - The second generation of methylenecyclopropane analogues of nucleosides 5a-5i and 6a-6i was synthesized and evaluated for antiviral activity. The 2,2 bis(hydroxymethyl)methylenecyclopropane (11) was converted to dibromo derivative 7 via acetate 12. Alkylation-elimination of adenine (16) with 7 afforded the Z/E mixture of acetates 17 + 18, which was deacetylated to give analogues 5a and 6a separated by chromatography. A similar reaction with 2-amino-6-chloropurine (19) afforded acetates 20 + 21 and, after deprotection and separation, isomers 5f and 6f. The latter served as starting materials for synthesis of analogues 5b, 5e, 5g 5i and 6b, 6e, 6g-6i. Alkylation-elimination of N(4)-acetylcytosine (22) with 7 afforded a mixture of isomers 5c + 6c which were separated via N(4)-benzoyl derivatives 23 and 24. Deprotection furnished analogues 5c and 6c. Alkylation of 2,4-bis(trimethylsilyloxy)-5-methylpyrimidine (25) with 7 led to bromo derivative 26. Elimination of HBr followed by deacetylation and separation gave thymine analogues 5d and 6d. The guanine Z-isomer 5b was the most effective against human and murine cytomegalovirus (HCMV and MCMV) with EC(50) = 0.27-0.49 microM and no cytotoxicity. The 6-methoxy analogue 5g was also active (EC(50) = 2.0-3.5 microM) whereas adenine Z-isomer 5a was less potent (EC(50) = 3.6-11.7 microM). Cytosine analogue 5c was moderately effective, but 2-amino-6-cyclopropylamino derivative 5e was inactive. All E-isomers were devoid of anti-CMV activity, and none of the analogues was significantly active against herpes simplex viruses (HSV-1 or HSV 2). The potency against Epstein-Barr virus (EBV) was assay-dependent. In Daudi cells, the E-isomers of 2-amino-6-cyclopropylamino- and 2,6-diaminopurine derivatives 6e and 6h were the most potent (EC(50) approximately 0.3 microM), whereas only the thymine Z-isomer 5d was active (EC(50) = 4.6 microM). Guanine Z derivative 5b was the most effective compound in H-1 cells (EC(50) = 7 microM). In the Z-series, the 2-amino-6-methoxypurine analogue 5g was the most effective against varicella zoster virus (VZV, EC(50) = 3.3 microM) and 2,6-diaminopurine 5h against hepatitis B virus (HBV, EC(50) = 4 microM). Adenine analogues 5a and 6a were moderately active as substrates for adenosine deaminase. PMID- 14736239 TI - SAR studies of dihydro-beta-agarofuran sesquiterpenes as inhibitors of the multidrug-resistance phenotype in a Leishmania tropica line overexpressing a P glycoprotein-like transporter. AB - A series of dihydro-beta-agarofuran sesquiterpenes isolated from the leaves of Maytenus cuzcoina (1-10) or semisynthetic derivatives (11-30) have been tested on a multidrug-resistant Leishmania tropica line overexpressing a P-glycoprotein like transporter to determine their ability to revert the resistance phenotype and to modulate intracellular drug accumulation. Almost all natural compounds showed potent reversal activity with different degrees of selectivity. Compounds 2, 7, and 8 are the most effective reversal agents tested against the multidrug resistance phenotype of Leishmania. Three-dimensional quantitative structure activity relationships using the comparative molecular similarity indices analysis (CoMSIA) were employed to characterize the steric (contribution of 5.4%), electrostatic (58.9%), lipophilic (10.0%), and hydrogen-bond-donor (13.3%) and -acceptor (7.5%) requirements of these sesquiterpenes as modulators at the P glycoprotein-like transporter. The most salient features of these requirements are the H-bond interaction between the substituents at the C-2 and C-6 positions with the receptor. PMID- 14736240 TI - Design of potent, selective, and orally bioavailable inhibitors of cysteine protease cathepsin k. AB - Osteoclast-mediated bone matrix resorption has been attributed to cathepsin K, a cysteine protease of the papain family that is abundantly and selectively expressed in osteoclast. Inhibition of cathepsin K could potentially be an effective method to prevent osteoporosis. Structure-activity studies on a series of reversible ketoamides based inhibitors of cathepsin K have led to identification of potent and selective compounds. Crystallographic studies have given insights into the mode of binding of these inhibitors. A series of ketoamides with varying P1 moieties were first synthesized to find an optimum group that would fit into the S1 subsite of the cysteine protease, cathepsin K. With a desired P1 group in place a variety of heterocyclic analogues in the P' region were synthesized to study their steric and electronic effects. In the process of exploring these P' heterocyclic variations, excellent selectivity was gained over other highly homologous cysteine proteases, including cathepsins L, S, and V. The favorable pharmacokinetic properties of some of these cathepsin K inhibitors in rats make them suitable for evaluation in rodent osteoporosis models. A representative cathepsin K inhibitor was shown to attenuate PTH stimulated hypercalcemia in the TPTX rat model. These inhibitors provide a viable lead series in the discovery of new therapies for the prevention and treatment of osteoporosis PMID- 14736241 TI - Design, synthesis, and in vitro biological evaluation of small molecule inhibitors of estrogen receptor alpha coactivator binding. AB - Nuclear receptors (NRs) complexed with agonist ligands activate transcription by recruiting coactivator protein complexes. In principle, one should be able to inhibit the transcriptional activity of the NRs by blocking this transcriptionally critical receptor-coactivator interaction directly, using an appropriately designed coactivator binding inhibitor (CBI). To guide our design of various classes of CBIs, we have used the crystal structure of an agonist bound estrogen receptor (ER) ligand binding domain (LBD) complexed with a coactivator peptide containing the LXXLL signature motif bound to a hydrophobic groove on the surface of the LBD. One set of CBIs, based on an outside-in design approach, has various heterocyclic cores (triazenes, pyrimidines, trithianes, cyclohexanes) that mimic the tether sites of the three leucines on the peptide helix, onto which are appended leucine residue-like substituents. The other set, based on an inside-out approach, has a naphthalene core that mimics the two most deeply buried leucines, with substituents extending outward to mimic other features of the coactivator helical peptide. A fluorescence anisotropy-based coactivator competition assay was developed to measure the specific binding of these CBIs to the groove site on the ER-agonist complex with which coactivators interact; control ligand-binding assays assured that their interaction was not with the ligand binding pocket. The most effective CBIs were those from the pyrimidine family, the best binding with K(i) values of ca. 30 microM. The trithiane- and cyclohexane-based CBIs appear to be poor structural mimics, because of equatorial vs axial conformational constraints, and the triazene-based CBIs are also conformationally constrained by amine-substituent-to-ring resonance overlap, which is not the case with the higher affinity alkyl-substituted pyrimidines. The pyrimidine-based CBIs appear to be the first small molecule inhibitors of NR coactivator binding. PMID- 14736242 TI - Design, synthesis, and biological activity of 4-[(4-cyano-2-arylbenzyloxy)-(3 methyl-3H-imidazol-4-yl)methyl]benzonitriles as potent and selective farnesyltransferase inhibitors. AB - A novel series of 4-[(4-cyano-2-arylbenzyloxy)-(3-methyl-3H-imidazol-4 yl)methyl]benzonitriles have been synthesized as selective farnesyltransferase inhibitors using structure-based design. X-ray cocrystal structures of compound 20-FTase-HFP and A313326-FTase-HFP confirmed our initial design. The decreased interaction between the aryl groups and Ser 48 in GGTase-I binding site could be one possible reason to explain the improved selectivity for this new series of FTase inhibitors. Medicinal chemistry efforts led to the discovery of compound 64 with potent cellular activity (EC(50) = 3.5 nM) and outstanding pharmacokinetic profiles in dog (96% bioavailable, 18.4 h oral t(1/2), and 0.19 L/(h x kg) plasma clearance). PMID- 14736243 TI - Synthesis, biological properties, and molecular modeling investigations of novel 3,4-diarylpyrazolines as potent and selective CB(1) cannabinoid receptor antagonists. AB - A series of novel 3,4-diarylpyrazolines was synthesized and evaluated in cannabinoid (hCB(1) and hCB(2)) receptor assays. The 3,4-diarylpyrazolines elicited potent in vitro CB(1) antagonistic activities and in general exhibited high CB(1) vs CB(2) receptor subtype selectivities. Some key representatives showed potent pharmacological in vivo activities after oral dosing in both a CB agonist-induced blood pressure model and a CB agonist-induced hypothermia model. Chiral separation of racemic 67, followed by crystallization and an X-ray diffraction study, elucidated the absolute configuration of the eutomer 80 (SLV319) at its C(4) position as 4S. Bioanalytical studies revealed a high CNS plasma ratio for the development candidate 80. Molecular modeling studies showed a relatively close three-dimensional structural overlap between 80 and the known CB(1) receptor antagonist rimonabant (SR141716A). Further analysis of the X-ray diffraction data of 80 revealed the presence of an intramolecular hydrogen bond that was confirmed by computational methods. Computational models and X-ray diffraction data indicated a different intramolecular hydrogen bonding pattern in the in vivo inactive compound 6. In addition, X-ray diffraction studies of 6 revealed a tighter intermolecular packing than 80, which also may contribute to its poorer absorption in vivo. Replacement of the amidine -NH(2) moiety with a NHCH(3) group proved to be the key change for gaining oral biovailability in this series of compounds leading to the identification of 80. PMID- 14736244 TI - Conformationally constrained analogues of diacylglycerol. 20. The search for an elusive binding site on protein kinase C through relocation of the carbonyl pharmacophore along the sn-1 side chain of 1,2-diacylglycerol lactones. AB - Previous studies with 1,2-diacylglycerol (DAG) lactones, which behave as high affinity ligands for protein kinase C (PK-C), have established the importance of maintaining intact the pharmacophore triad of two carbonyl moieties (sn-1 and sn 2) and the primary alcohol. In addition, docking studies of DAG-lactones into an empty C1b receptor of PK-Cdelta (as it appears in complex with phorbol 13-O acetate) have revealed that in either of the two possible binding alternatives (sn-1 or sn-2) only one carbonyl group of the DAG-lactone is involved in binding. Therefore, the unknown receptor for the orphaned carbonyl appears to lie outside the boundaries of this binary complex, possibly residing at the membrane or near the membrane-protein interface. A strategy to locate the optimal location of the unengaged carbonyl was conceived by utilizing a small group of DAG-lactones (1-4) with a highly branched chain adjacent to the sn-2 carbonyl such that sn-2 binding is favored. With these compounds, various locations of the sn-1 carbonyl along the side chain were tested for their binding affinity for PK-C. The results indicate that the location of the side chain sn-1 carbonyl in a DAG-lactone must have perfect mimicry to the sn-1 carbonyl of the parent DAG for it to display high binding affinity. A proposed model from this work is that the missing pharmacophore in the ternary complex, which includes the membrane, is close to the membrane-protein interface. PMID- 14736245 TI - Structure-activity relationships of N-acyl pyrroloquinolone PDE-5 inhibitors. AB - The discovery of the potent and selective PDE-5 inhibitory activity of a pyrroloquinolone scaffold prompted us to explore the SAR of its acyl derivatives. During the course of these studies, three structural series were found with K(i) values for PDE-5 in the subnanomolar range. Systematic modification of one of these leads produced a compound with excellent selectivity for PDE-5 over other phosphodiesterases and oral bioavailability of 15% in male rats. This compound also displayed in vivo efficacy in an anesthetized canine model of erection when dosed intravenously. PMID- 14736246 TI - Synthesis and biological evaluation of 2,3,5-substituted [1,2,4]thiadiazoles as allosteric modulators of adenosine receptors. AB - A number of 2,3,5-substituted [1,2,4]thiadiazole analogues of SCH-202676 (N-(2,3 diphenyl[1,2,4]thiadiazole-5(2H)-ylidene)methanamine, 7a) were synthesized and tested as potential allosteric modulators of adenosine receptors. All compounds were capable of displacing the binding of the radiolabeled agonist [(3)H]CCPA to human A(1) adenosine receptors, whereas modest and varying effects were observed on the binding of [(3)H]DPCPX, a radiolabeled antagonist for this receptor subtype. Four compounds, 7a (SCH-202676), 7k (LUF5792), 7l (LUF5794), and 8e (LUF5789), were selected for more detailed characterization. They all proved allosteric inhibitors of agonist binding, with 7k being most potent, whereas their effects on antagonist binding were more ambiguous. Subsequently, experiments were done on human adenosine A(2A) and A(3) receptors. Compounds 7a and 7l displayed peculiar displacement characteristics of both radiolabeled agonist and antagonist binding to A(2A) receptors, whereas 7a showed some activity on A(3) receptors. PMID- 14736247 TI - Inhibitors of multiple mutants of Plasmodium falciparum dihydrofolate reductase and their antimalarial activities. AB - Novel analogues of pyrimethamine (Pyr) and cycloguanil (Cyc) have been synthesized and tested as inhibitors of Plasmodium falciparum dihydrofolate reductase carrying triple (N51I+C59R+S108N, C59R+S108N+I164L) and quadruple (N51I+C59R+S108N+I164L) mutations responsible for antifolate resistance. The inhibitors were designed to avoid steric clash of the p-Cl group of the inhibitors with the side chain of Asn108, augmented by additional mutations of the resistant mutants. Cycloguanil derivatives were also designed to avoid steric clash with the side chain of Val16 in the A16V+S108T mutant. Many compounds have inhibition constants (K(i)) at the low nanomolar level against the mutant enzymes and a number have good antimalarial activities against resistant P. falciparum parasites bearing multiple mutations in the S108N series and A16V+S108T mutant enzymes. These compounds in the Pyr and Cyc series exhibit low and moderate cytotoxicity to nontumor (Vero) and tumor (KB, BC) cell lines. Some of these inhibitors are therefore potential candidates for further development as antimalarials. PMID- 14736248 TI - Design, synthesis, and biological evaluation of thio-containing compounds with serum HDL-cholesterol-elevating properties. AB - A novel series of substituted sulfanyldihydroimidazolones (1) that modulates high density lipoprotein cholesterol (HDL-C) has been reported to have HDL-elevating properties in several animal models. Concerns about the chemical and metabolic stability of 1 directed us to explore the structure-activity relationship (SAR) of a related series of substituted thiohydantoins (2). Expansion of the scope of the thiohydantoin series led to exploration of compounds in related thio containing ring systems 3-7 and the N-cyanoguanidine derivative 8. Compounds were tested sequentially in three animal models to assess their HDL-C elevating efficacy and safety profiles. Further evaluation of selected compounds in a dose response paradigm culminated in the identification of compound 2.39 as a candidate compound for advanced preclinical studies. PMID- 14736249 TI - Total synthesis and antitubulin activity of c10 analogues of cryptophycin-24. AB - The unsubstituted, 3'-Cl, 4'-C1, and 3',4'-diCl C10 analogues of cryptophycin-24 were prepared via total synthesis and tested in vitro for cytotoxicity against MCF-7 and multi-drug-resistant MCF-7/ADR breast cancer cell lines and in a tubulin assembly assay. The ED(50) values ranged from 7.2 to 15.8 microM in the tubulin assay and from 0.05 to 3.4 nM in the cell assays. The presence of a 3'-C1 and/or 4'-C1 substituent on the C10 phenyl ring increased cytotoxicity in the MCF 7 cell line compared to the unsubstituted phenyl ring. The most potent compound in this series possessed a 3'-C1 substituent on the C10 phenyl ring. The 3'-C1 analogue had ED(50) values of 50 and 580 pM in the MCF-7 and MCF-7/ADR cell lines, respectively. Its activity was very similar to the parent compound cryptophycin-24. Substitution of the 4'-MeO group in cryptophycin-24 with a 4'-C1 moiety did not significantly affect cytotoxicity against MCF-7 and MCF-7/ADR cells compared to the parent compound. These results demonstrated that the 4'-MeO group in cryptophycin-24 is not essential and can be replaced with 3'-C1 or 4'-C1 substituents. PMID- 14736250 TI - Potent and selective conformationally restricted neuronal nitric oxide synthase inhibitors. AB - Selective inhibition of the isoforms of nitric oxide synthase (NOS) in pathologically elevated synthesis of nitric oxide has great therapeutic potential. We previously reported nitroarginine-containing dipeptide amides and some peptidomimetic analogues as potent and selective inhibitors of neuronal NOS (nNOS). Here we report conformationally restricted dipeptides derived from the dipeptide L-Arg(NO2)-L-Dbu-NH2 (8). The selectivities for nNOS over endothelial NOS and inducible NOS of the most potent nNOS inhibitor (10a) among these compounds are comparable to that of the parent compound. An unsubstituted amide bond is necessary for potency against nNOS. The stereochemistry of compound 10a was optimum for potency and selectivity and thus provides the binding conformation of the parent compound with nNOS. PMID- 14736251 TI - Synthesis of nitro esters of prednisolone, new compounds combining pharmacological properties of both glucocorticoids and nitric oxide. AB - Glucocorticoids (GC) are widely used in therapy for their many pharmacological properties including antiinflammatory and immunosuppressive actions. However, their use over long periods is hampered by a number of severe side effects. Given the biological properties of nitric oxide (NO) and previous experience with nonsteroidal antiinflammatory agents, we synthesized new chemical entities combining both NO and GC properties. Here we report the synthesis of nitro esters of prednisolone obtained through the esterification, with different linkers, on the hydroxy group at C-21 position of the corticosteroid structure. The alkyl chain, as of the nitrooxy derivative (2), or aromatic linkers, as of o-, m-, and p-nitrooxymethylbenzoate derivatives (3-5), respectively, furnish stable compounds that release NO and inhibit the GC receptors in biological assays. To improve solubility we introduced a more water-soluble linker such as the nitrooxyalkylpiperidine or -piperazine group (6-9). Also these compounds retained properties of both NO and prednisolone. Compound 5 (NCX 1015) was selected for its better profile: enhanced antiinflammatory properties and reduced side effects compared with prednisolone. NCX 1015 is currently under preclinical development. PMID- 14736252 TI - 4-[5-Fluoro-3-[4-(2-methyl-1H-imidazol-1-yl)benzyloxy]phenyl]-3,4,5,6- tetrahydro 2H-pyran-4-carboxamide, an orally active inhibitor of 5-lipoxygenase with improved pharmacokinetic and toxicology characteristics. AB - Described herein are structure-activity relationships (SARs) of 4-[5-fluoro-3-[4 (2-methyl-1H-imidazol-1-yl)benzyloxy]-phenyl]-4-methoxy-3,4,5,6-tetrahydro-2H pyran (1, CJ-12,918), an imidazole 5-lipoxygenase (5-LO) inhibitor. When 1 was tested in preclinical studies, cataract formation was observed in rats; however, this compound was metabolized extensively in vivo and showed low systemic exposure. To eliminate this side effect and enhance bioavailability, structural modification was focused on replacing the methoxy group of 1 by modulating lipophilicity (i.e., predicted log D at pH 7.4). The SARs led to the discovery of 4-[5-fluoro-3-[4-(2-methyl-1H-imidazol-1-yl)benzyloxy]phenyl]-3,4,5,6-tetrahydro 2H-pyran-4-carboxamide (10, CJ-13,454), which was less lipophilic by 1.2 log D units and showed in vivo potency (ED(50) = 4-9 mg/kg) equipotent to 1. Enhanced metabolic stability resulted in fewer in vivo metabolites, as well as improved bioavailability and a better toxicological profile. Thus, 10 was found to be a more practical lead for an orally active 5-LO inhibitor. PMID- 14736253 TI - A new aliphatic amino prodrug system for the delivery of small molecules and proteins utilizing novel PEG derivatives. AB - A new amino PEG prodrug system, based entirely on aliphatic structures, has been designed using ester derivatives easily synthesized from N-modified bis-N-2 hydroxyethylglycinamides. Hydrolysis of the various promoiety bonds, in vivo, regenerated amine in a predictable manner. Thus, a novel new methodology for controlled release of amino-containing drugs, peptides, and proteins has been accomplished. This work demonstrates the usefulness of a PEG prodrug strategy that results in solubilization of insoluble amino-containing drugs and provides prodrugs with relatively long circulating half-lives. It can be appreciated that this novel system should also be applicable for nonpolymer-containing prodrugs as well. PMID- 14736254 TI - Structure-activity study on the Phe side chain arrangement of endomorphins using conformationally constrained analogues. AB - Endomorphins-1 and -2 were substituted with all the beta-MePhe stereoisomers in their Phe residues to generate a conformationally constrained peptide set. This series of molecules was subjected to biological assays, and for beta-MePhe(4) endomorphins-2, a conformational analysis was performed. Incorporation of (2S,3S) beta-MePhe(4) resulted in the most potent analogues of both endomorphins with enhanced enzymatic stability. Their micro opioid affinities were 4-times higher than the parent peptides, they stimulated [(35)S]GTPgammaS binding, and they were found to be full agonists. NMR experiments revealed that C-terminal (2S,3S)-beta MePhe in endomorphin-2 strongly favored the gauche (-) spatial orientation which implies the presence of the chi(1) = -60 degrees rotamer of Phe(4) in the binding conformer of endomorphins. Our results emphasize that the appropriate orientation of the C-terminal aromatic side chain of endomorphins is substantial for binding to the micro opioid receptor. PMID- 14736255 TI - Synthesis and structure-activity relationships of novel arylpiperazines as potent and selective agonists of the melanocortin subtype-4 receptor. AB - The melanocortin receptors have been implicated as potential targets for a number of important therapeutic indications, including inflammation, sexual dysfunction, and obesity. We identified compound 1, an arylpiperazine attached to the dipeptide H-d-Tic-d-p-Cl-Phe-OH, as a novel melanocortin subtype-4 receptor (MC4R) agonist through iterative directed screening of nonpeptidyl G-protein coupled receptor biased libraries. Structure-activity relationship (SAR) studies demonstrated that substitutions at the ortho position of the aryl ring improved binding and functional potency. For example, the o-isopropyl-substituted compound 29 (K(i) = 720 nM) possessed 9-fold better binding affinity compared to the unsubstituted aryl ring (K(i) = 6600 nM). Sulfonamide 39 (K(i) = 220 nM) fills this space with a polar substituent, resulting in a further 2-fold improvement in binding affinity. The most potent compounds such as the diethylamine 44 (K(i) = 60 nM) contain a basic group at this position. Basic heterocycles such as the imidazole 50 (K(i) = 110 nM) were similarly effective. We also demonstrated good oral bioavailability for sulfonamide 39. PMID- 14736256 TI - Anti-AIDS agents. 52. Synthesis and anti-HIV activity of hydroxymethyl (3'R,4'R) 3',4'-di-O-(S)-camphanoyl-(+)-cis-khellactone derivatives. AB - To enhance the water solubility and oral bioavailability of DCK analogues, 12 new mono- and disubstituted (3'R,4'R)-3',4'-di-O-(S)-camphanoyl-(+)-cis-khellactone (DCK) analogues were synthesized and evaluated for inhibition of HIV-1 replication in H9 lymphocytes. 3-Hydroxymethyl-4-methyl-DCK (4c) exhibited significant anti-HIV activity in H9 lymphocytes and primary peripheral blood mononuclear cells with EC(50) values of 0.004 and 0.024 microM, respectively. Although this compound was not as potent as 4-methyl-DCK (2) and 3-bromomethyl-4 methyl-DCK (4a), it provides increased water solubility and possible linkage to other moieties. Of particular note, 4c exhibits moderate oral bioavailability (15%) when administered as a carboxymethylcellulose suspension to rats, whereas 2 is not orally bioavailable in the same formulation. Further studies on mechanism of action suggest that 4c inhibits the production of double-stranded viral DNA from the single-stranded DNA intermediate. In addition, 4a is the most potent compound in this series of new analogues, with EC(50) and TI values of 0.00011 microM and 189,600, respectively. Thus, further modification at the 3-position of the coumarin ring can improve the potency of new DCK analogues. PMID- 14736257 TI - Platinum(IV) complex with adamantylamine as nonleaving amine group: synthesis, characterization, and in vitro antitumor activity against a panel of cisplatin resistant cancer cell lines. AB - Procedure of the synthesis is described for new platinum(IV) drug LA-12 [(OC-6 43)-bis(acetato)(1-adamantylamine)amminedichloroplatinum(IV)]. The X-ray diffraction analysis shows that the structure is created by molecules with octahedral arrangement of ligands around a platinum atom and contains one H(2)O molecule that is not a part of the coordination sphere of platinum. This new drug is more reactive with glutathione than cisplatin and is lacking cross-resistance with cisplatin as proven on the panel of cancer cell lines. PMID- 14736258 TI - Performance of 3D-database molecular docking studies into homology models. AB - The performance of docking studies into protein active sites constructed by homology model building was investigated using CDK2 and factor VIIa screening data sets. When the sequence identity between model and template near the binding site area is greater than approximately 50%, roughly 5 times more active compounds are identified than would be found randomly. This performance is comparable to docking to crystal structures. PMID- 14736261 TI - A rosette by any other name... PMID- 14736262 TI - Pathologic quiz case: a 59-year-old man with a 2-week history of right iliac fossa pain. Nonsteroidal anti-inflammatory drug-induced colonic ulcer. PMID- 14736263 TI - Pathologic quiz case: a 69-year-old asymptomatic man with a scrotal mass. Atypical (symplastic or bizarre) leiomyoma of the scrotum. PMID- 14736264 TI - Pathologic quiz case: bilateral apical lung masses in an autopsy patient. Pulmonary apical cap. PMID- 14736265 TI - Pathologic quiz case: headache in an 8-year-old child. Primary amebic meningoencephalitis due to Naegleria fowleri. PMID- 14736266 TI - Pathologic quiz case: a 40-year-old woman with an unusual uterine tumor. Uterine angiolipoleiomyoma with focal atypical leiomyoma. PMID- 14736267 TI - Pathologic quiz case: a solitary breast nodule in an elderly woman. Fasciitis ossificans of the breast. PMID- 14736268 TI - Oncocytic biliary cystadenocarcinoma: a case report and review of the literature. AB - We report an unusual case of biliary cystadenocarcinoma with oncocytic differentiation. The patient was a 43-year-old woman who presented with right upper quadrant pain. Imaging revealed a 16 x 10 x 10-cm, heterogenous, right hepatic mass with extension into the right atrium. Surgical resection revealed a papillary neoplasm of malignant cells with atypical hyperchromatic nuclei and prominent nucleoli lining fibrovascular cores. Mesenchymal stroma was not present. The majority of the epithelial cells had abundant eosinophilic granular cytoplasm, consistent with oncocytic differentiation. There was extensive stromal and hepatic parenchymal invasion. Immunohistochemical staining revealed a "biliary pattern" of cytokeratin subset immunoreactivity, with positivity for cytokeratin 7 and an absence of staining with cytokeratin 20. The tumor was negative for mucin, carcinoembryonic antigen, alpha-fetoprotein, calretinin, CD31, and chromogranin. There was granular cytoplasmic staining with phosphotungstic acid hematoxylin, consistent with the presence of abundant mitochondria. Electron microscopy revealed abundant mitochondria within the neoplastic cells. This case is quite unusual because female patients only rarely lack the characteristic ovarian-like mesenchymal stroma of biliary cystadenomas/cystadenocarcinomas. Furthermore, to our knowledge, oncocytic differentiation in this neoplasm has been reported previously on only 2 occasions. The biologic behavior and prognostic significance, if any, of the lack of mesenchymal stroma in female patients or the presence of oncocytic differentiation remains to be further elucidated as more of these cases are described. PMID- 14736269 TI - Sebaceous differentiation in salivary glands. PMID- 14736270 TI - Aberrant nuclear projections of neutrophils in trisomy 13. PMID- 14736271 TI - Pure erythroid leukemia. PMID- 14736272 TI - Pathologic quiz case: an 11-year-old boy with acute-onset right lower abdominal pain. Typhilitis (neutropenic enterocolitis). PMID- 14736273 TI - Pathologic quiz case: sacral/pelvic mass with lower back pain in a 36-year-old man. Immature plasmacytoma. PMID- 14736274 TI - Pathologic quiz case: a rapidly increasing breast mass in a postmenopausal woman. Malignant adenomyoepithelioma. PMID- 14736275 TI - Pathologic quiz case: renal impairment after bone marrow transplantation. Bone marrow transplant nephropathy. PMID- 14736276 TI - Collagenous gastritis: an unusual association with profound weight loss. AB - Collagenous gastritis is a distinctive disorder characterized by thickening of the subepithelial collagen layer in the gastric mucosa. Although this entity was recognized in 1989, its etiology, pathogenesis, and clinicopathologic features remain poorly understood because of its rarity. An unusual case of collagenous gastritis was observed in a 37-year-old man who presented with profound weight loss, a feature that has not previously been emphasized. PMID- 14736277 TI - Multilocular thymic cyst associated with mature mediastinal teratoma: a report of 2 cases. AB - Acquired thymic cysts are multilocular and show florid xanthomatous and myofibroblastic inflammation. They usually occur in association with mediastinal neoplasms, systemic autoimmune diseases, or trauma. We describe 2 cases (in a 12 year-old girl and an 11-year-old boy) of acquired thymic cysts occurring in association with cystic teratomas, an association that to our knowledge has not been described previously in the literature. PMID- 14736278 TI - Fibrolamellar hepatocellular carcinoma occurring 5 years after hepatocellular adenoma in a 14-year-old girl: a case report with comparative genomic hybridization analysis. AB - An unusual case of fibrolamellar carcinoma of the liver developed 5 years after removal of a hepatocellular adenoma in a 14-year-old girl belonging to a family with Carney syndrome. Both tumors were studied by light and electron microscopy, flow cytometry, and comparative genomic hybridization. The first tumor, removed at the age of 9, was a bulky well-circumscribed liver mass composed of large eosinophilic cells with a focal pseudoglandular pattern but without cytologic atypia or sclerosis. A diagnosis of hepatocellular adenoma was rendered. Five years later, another hepatic tumor was removed from the right lobe. Microscopic examination revealed polygonal cells, each with a large amount of eosinophilic cytoplasm and a round nucleus with a conspicuous nucleolus. These cells were arranged in nests and strands and separated by bands of dense fibrous tissue, leading to a diagnosis of fibrolamellar carcinoma. Comparative genomic hybridization analysis revealed no genetic alteration in the adenoma; however, several chromosomal aberrations (loss of chromosome regions 1p and 4p and gains of chromosome regions 6q, 13q, and Xq) were detected in the fibrolamellar carcinoma. To our knowledge, this is the first report of an association between hepatocellular adenoma and fibrolamellar carcinoma. PMID- 14736279 TI - Advanced gastric carcinoma with a complete intestinal metaplasia phenotype associated with early intestinal-type carcinoma. AB - An unusual case of synchronous gastric carcinomas occurred in a 28-year-old man with a family history of gastric disease. Two tumor foci were identified: a well differentiated advanced carcinoma with the phenotypic properties of complete intestinal metaplasia and an early intestinal-type carcinoma. Histochemical and immunohistochemical stains to demonstrate complete intestinal metaplasia, ie, Alcian blue pH 2.5/periodic acid-Schiff, high iron diamine/Alcian blue pH 2.5, CD10, and MUC2, were all positive in the advanced adenocarcinoma. Of all markers used, only high iron diamine/Alcian blue pH 2.5 and Alcian blue pH 0.5 were positive in the early carcinoma. In these cases, mistakes frequently are made during examination of endoscopic biopsies. Fortunately, the advanced adenocarcinoma was low grade (the patient has shown no signs of disease at 6 years postsurgery). Histopathologic, histochemical, and immunohistochemical findings suggest that an extensive substrate of complete intestinal metaplasia (corpus) and of complete and incomplete intestinal metaplasia (antrum) can be associated with two independent tumors with different phenotypes. PMID- 14736280 TI - Jejunal gastrointestinal stromal tumor: a cause of obscure gastrointestinal bleeding. AB - In cases of obscure gastrointestinal bleeding, when a source for blood loss is not apparent from examination of the colon and upper gastrointestinal tract, the small bowel usually becomes the focus of investigation. A tumor with interesting pathologic features was found in a patient who presented with recurrent episodes of massive obscure gastrointestinal hemorrhage. This case highlights the importance of considering small intestinal tumors as the likely cause of obscure gastrointestinal hemorrhage in young patients and how a noninvasive test, eg, abdominal computed tomography scan, might obviate the need for more invasive testing. PMID- 14736281 TI - Follicular lymphoma with a burkitt translocation--predictor of an aggressive clinical course: a case report and review of the literature. AB - Follicular lymphoma is an indolent lymphoma characterized by the (14;18) translocation, which leads to aberrant expression of Bcl-2. Translocations involving 8q24 are most commonly associated with Burkitt lymphoma and result in c Myc overexpression. We report a case of follicular lymphoma of predominant small cleaved-cell type (grade 1) associated with both a t(14;18)(q32;q21) and a t(8;22)(q24;q11). The 8q24 translocation predicted an aggressive clinical course, as the lymphoma transformed into acute lymphoblastic leukemia within a year of initial diagnosis. Routine cytogenetic analysis is recommended at initial diagnosis of follicular lymphoma to better identify abnormalities that may predict prognosis and influence therapy. PMID- 14736282 TI - Use of polymerase chain reaction to diagnose tuberculous arthritis from joint tissues and synovial fluid. AB - CONTEXT: Tuberculosis of the joints and bones is a significant worldwide problem, often leading to joint and bone destruction. The diagnosis of this disease manifestation is difficult. OBJECTIVE: To assess the role of conventional diagnostics compared to polymerase chain reaction applied to samples obtained at arthroscopy. DESIGN: This was an open observational study that was blinded to the microbiologist, histopathologist, and molecular biologist responsible for assessing the main outcome measures. PATIENTS: Seven patients (8 samples) with joint and bone tuberculosis and 14 patients (16 samples) with nontuberculous joint and bone disease. INTERVENTION: Arthroscopic examination and tissue sample collection. MAIN OUTCOME MEASURES: Mycobacterium tuberculosis staining, culture, and histopathologic assessment of caseating granulomas vs polymerase chain reaction. RESULTS: Polymerase chain reaction was positive in all cases of true tuberculosis and falsely identified 2 samples as positive, both however, in patients who had lung tuberculosis in the past. CONCLUSIONS: Conventional bacteriological methods for demonstration of M tuberculosis are not very sensitive and can be time-consuming. Polymerase chain reaction of arthroscopically obtained joint tissue biopsies appears promising in the early diagnosis of tuberculous arthritis. PMID- 14736283 TI - Analysis of deaths during the severe acute respiratory syndrome (SARS) epidemic in Singapore: challenges in determining a SARS diagnosis. AB - CONTEXT: An outbreak of severe acute respiratory syndrome (SARS), an infectious disease attributed to a novel coronavirus, occurred in Singapore during the first quarter of 2003 and led to 204 patients with diagnosed illnesses and 26 deaths by May 2, 2003. Twenty-one percent of these patients required admission to the medical intensive care unit. During this period, the Center for Forensic Medicine, Health Sciences Authority, Singapore, performed a total of 14 postmortem examinations for probable and suspected SARS. Of these, a total of 8 were later confirmed as SARS infections. OBJECTIVE: Our series documents the difficulties encountered at autopsy during the initial phases of the SARS epidemic, when the pattern of infection and definitive diagnostic laboratory criteria were yet to be established. DESIGN: Autopsies were performed by pathologists affiliated with the Center for Forensic Medicine, Health Sciences Authority, Singapore. Tissue was accessed and read at the Tan Tock Seng Hospital, Singapore, and at the Armed Forces Institute of Pathology, Washington, DC. Autopsy tissue was submitted to the Virology Department, Singapore General Hospital, for analysis, and in situ hybridization for the SARS coronavirus was carried out at the National Institute of Infectious Diseases, Tokyo, Japan. RESULTS: Thirteen of 14 patients showed features of diffuse alveolar damage. In 8 patients, no precipitating etiology was identified, and in all of these patients, we now have laboratory confirmation of coronavirus infection. Two of the 8 patients presented at autopsy as sudden unexpected deaths, while the remaining 6 patients had been hospitalized with varying lengths of stay in the intensive care unit. In 3 patients, including the 2 sudden unexpected deaths, in situ hybridization showed the presence of virally infected cells within the lung. In 4 of the 8 SARS patients, pulmonary thromboemboli were also recognized on gross examination, while one patient had marantic cardiac valvular vegetations. CONCLUSIONS: It is unfortunate that the term atypical pneumonia has been used in conjunction with SARS. Although nonspecific by itself, the term does not accurately reflect the underlying dangers of viral pneumonia, which may progress rapidly to acute respiratory distress syndrome. We observed that the clinical spectrum of disease as seen in our autopsy series included sudden deaths. This is a worrisome finding that illustrates that viral diseases will have a spectrum of clinical presentations and that the diagnoses made for such patients must incorporate laboratory as well as clinical data. PMID- 14736284 TI - Clinicopathologic study of hepatic Penicillium marneffei in Northern Thailand. AB - CONTEXT: Penicillium marneffei, an opportunistic fungus, is endemic in Southeast Asia, especially in human immunodeficiency virus-infected individuals living in northern Thailand. OBJECTIVE: We present the results of a clinicopathologic study of hepatic penicilliosis among human immunodeficiency virus/acquired immunodeficiency syndrome patients. DESIGN: A search of liver biopsies in one institution from 1998 to 1999 identified 30 cases of penicilliosis. RESULTS: Histologically, hepatic lesions could be classified into 1 of 3 patterns: diffuse, granulomatous, and mixed. The diffuse pattern showed a diffuse infiltration of foamy macrophages that contained numerous P marneffei. The granulomatous pattern showed a formation of multiple granulomata with various degrees of inflammatory cell infiltration. The mixed pattern showed features intermediate between the diffuse and granulomatous patterns. Liver function tests of the 3 pathologic pattern groups were evaluated, but there were no statistically significant differences in aspartate aminotransferase, alanine aminotransferase, or alkaline phosphatase levels among the various histologic groups. CONCLUSION: To our knowledge, this is the largest series to date that documents the liver pathology that results from this pathogen. We hypothesize that the histologic patterns seen on biopsy reflect the level of the host's immunity. Hence, in addition to a diagnosis of penicilliosis, a liver biopsy may also provide an assessment of the host's immune status, whereas liver function tests do not. PMID- 14736285 TI - Allelic loss of the active X chromosome during bladder carcinogenesis. AB - CONTEXT: Previous studies have shown that loss of the X chromosome is involved in the carcinogenesis of certain human malignancies. OBJECTIVE: To determine whether X-linked allelic losses occur during bladder tumorigenesis and whether such losses involve the active or the inactive X chromosome. DESIGN: We analyzed the deletion status of the X-linked human androgen receptor gene locus in 6 female patients who underwent radical cystectomies for muscle-invasive urothelial carcinoma of the urinary bladder. Four patients had coexisting urothelial carcinoma in situ. Analysis for inactivation of the X chromosome was carried out in parallel. RESULTS: Three cases were informative. Invasive tumor samples showed loss of heterozygosity involving the active allele at the androgen receptor locus in all 3 positive cases, whereas carcinoma in situ showed nonrandom X chromosome inactivation but not allelic deletion. CONCLUSIONS: Our data suggest that allelic loss of the activated X chromosome is involved in bladder carcinogenesis and cancer progression. PMID- 14736286 TI - Flow cytometric evaluation of posttransplant B-cell lymphoproliferative disorders. AB - CONTEXT: Posttransplant B-cell lymphoproliferative disorders (PTLDs) constitute a heterogeneous group that includes hyperplastic and unique polymorphic lesions at one end of the spectrum and monomorphic lymphoid proliferations indistinguishable morphologically from conventional B-cell non-Hodgkin lymphomas (NHLs) at the other end. Almost all the PTLDs are of B-cell origin, with only rare examples of T-cell phenotype described. Despite a plethora of information available on the morphologic spectrum, pathogenetic role of Epstein-Barr virus, and various treatment options, a detailed flow cytometric immunophenotypic evaluation of PTLDs is largely lacking. OBJECTIVE: To evaluate the immunophenotypic profiles of various PTLDs using multiparameter flow cytometric analysis to compare and contrast with conventional de novo B-cell lymphoproliferative disorders and to identify any immunophenotypic patterns useful in diagnosis. DESIGN: We retrospectively analyzed data on the immunophenotype of 25 cases of pediatric and adult PTLD (12 cases of monomorphic PTLD [m-PTLD] and 13 cases of polymorphic PTLD [p-PTLD]) using multiparameter flow cytometry in addition to routine morphologic and immunohistochemical evaluation. The flow cytometric immunophenotypic data were also compared and contrasted with 334 cases of various de novo B-cell NHLs during the same period as a control group. RESULTS: We observed a much higher incidence of lack of surface immunoglobulin light chains and CD20 expression in B-cell PTLDs using multiparameter flow cytometry in comparison with de novo B-cell NHL as a group (with the exception of small lymphocytic lymphoma). Four (16%) of 25 cases of PTLD (3 m-PTLD and 1 p-PTLD) showed almost complete lack (CD20%/CD19% ratio < 1:9) of CD20 expression in contrast to only 8 ( approximately 2%) of 334 cases of de novo B-cell NHL (P =.007). Several other cases of both m-PTLD and p-PTLD also showed partial and dim expression of CD20. Nine (36%) of 25 cases, including 5 cases of m-PTLD and 4 of p-PTLD, showed either an almost complete lack (light chains%/CD19% ratio < 1:9) or significant loss (>50% loss) of surface immunoglobulin light chains in contrast to less than 5% incidence of light-chain negativity in conventional de novo B-cell NHL. Immunoglobulin light-chain clonality was observed in 9 cases (5 m-PTLD and 4 p-PTLD). Seven cases (5 p-PTLD and 2 m-PTLD) had polyclonal expression of immunoglobulin kappa and lambda light chains. The m-PTLD showed expression patterns of CD5, CD10, and CD23 similar to their de novo counterparts. CONCLUSIONS: Both polymorphic and monomorphic PTLDs show a higher incidence of lack of CD20 and surface immunoglobulin light-chain expression. The lack of CD20 expression in these lesions may have therapeutic implications, since anti-CD20 antibody has increasingly become an important modality in the treatment of B-cell lymphoproliferative disorders, including posttransplant disorders. PMID- 14736287 TI - Reflectance spectroscopy of clotting blood: a description of the time-dependent behavior. AB - CONTEXT: Research into whether cyclooxygenase-2 (COX-2) inhibitors affect thrombosis has been hampered by the lack of a specific assay. Erythrocytes modulate the effect of aspirin on platelets, which suggests that tests of whole blood clotting may be more sensitive. OBJECTIVES: To determine whether reflectance spectroscopy of clotting blood generates useful information about coagulation and whether it shows an effect of COX-2 inhibitors. DESIGN: A survey of 14 adults examined the range of phenomena demonstrated by reflectance spectroscopy. These phenomena were compared before and after treatment with a COX 2 inhibitor in 4 subjects. SETTING: Out-patient clinic. MAIN OUTCOME MEASURE: Reflected light intensity was measured from blood as it clotted in a cuvette thermostated at 37 degrees C. RESULTS: The survey of healthy adults showed that the time course of reflected light intensity is similar at all wavelengths and may be divided into 4 stages: a monotonic decrease, a sigmoidal increase, a linear region, and a terminal phase. Clot formation as determined by tube inversion occurs at the transition between the first and second phases; the sigmoidal increase cannot be due to fibrin polymerization. The terminal phase coincides with clot retraction. Similar results are obtained in native whole blood and in recalcified citrated blood. Cyclooxygenase-2 inhibitors have an intrinsic effect on the sigmoidal increase ex vivo (P <.001). CONCLUSIONS: Reflectance spectroscopy generates unique information about clotting blood. It is feasible to use anticoagulated blood to elucidate the events underlying the time course and to investigate the effects of COX-2 inhibitors. PMID- 14736288 TI - Use of physician extenders in surgical pathology practice. AB - CONTEXT: Use of a variety of nonphysician personnel for surgical pathology gross examination is generally known to be increasing, although detailed information regarding nonphysician use is currently unavailable. OBJECTIVE: To measure and describe the use of nonphysician personnel for surgical pathology gross examination in order to gain a better understanding of the current surgical pathology workforce. DESIGN: A voluntary, mailed questionnaire containing items related to the use of multiple nonphysician personnel types in surgical pathology was distributed to (1) a cross-sectional sample (n = 968) of US pathologists and (2) a purposive sample of pathologist directors of surgical and/or anatomic pathology (n = 77) located at teaching institutions. Responses were analyzed using descriptive statistics, correlation analyses, the chi2 test, and 1-way analysis of variance. Staffing ratios were calculated for multiple nonphysician personnel types. RESULTS: The overall response rate was 22% (n = 225). Of the US sample, 56% of respondents reported using nonphysician laboratory personnel to perform gross examinations, compared with 91% of the directors' sample. The most frequently reported personnel type for both samples was pathologists' assistants, but multiple other personnel types were used as well. Significant associations existed between certain practice types and personnel types used, as well as differences in the scope of responsibilities between personnel types. Calculated staffing ratios were variable across personnel types and were highest for pathologists' assistants. CONCLUSIONS: The use of a variety of nonphysician laboratory personnel for surgical pathology gross examination is common, particularly in academic pathology practice. Further studies are needed to examine the impact of physician extenders on laboratory efficiency and quality of care. PMID- 14736289 TI - Biochemical markers of myocardial injury test turnaround time: a College of American Pathologists Q-Probes study of 7020 troponin and 4368 creatine kinase-MB determinations in 159 institutions. AB - CONTEXT: Rapid diagnosis of acute myocardial infarction in patients presenting to emergency departments (EDs) with chest pain may determine the types, and predict the outcomes of, the therapy those patients receive. The amount of time consumed in establishing diagnoses of acute myocardial infarction may depend in part on that consumed in the generation of the blood test results measuring myocardial injury. OBJECTIVE: To determine the normative rates of turnaround time (TAT) for biochemical markers of myocardial injury and to examine hospital and laboratory practices associated with faster TATs. DESIGN: Laboratory personnel in institutions enrolled in the College of American Pathologists Q-Probes Program measured the order-to-report TATs for serum creatine kinase-MB and/or serum troponin (I or T) for patients presenting to their hospital EDs with symptoms of acute myocardial infarction. Laboratory personnel also completed detailed questionnaires characterizing their laboratories' and hospitals' practices related to testing for biochemical markers of myocardial injury. ED physicians completed questionnaires indicating their satisfaction with testing for biochemical markers of myocardial injury in their hospitals. SETTING: A total of 159 hospitals, predominantly located in the United States, participating in the College of American Pathologists Q-Probes Program. RESULTS: Most (82%) laboratory participants indicated that they believed a reasonable order-to-report TATs for biochemical markers of myocardial injury to be 60 minutes or less. Most (75%) of the 1352 ED physicians who completed satisfaction questionnaires believed that the results of tests measuring myocardial injury should be reported back to them in 45 minutes or less, measured from the time that they ordered those tests. Participants submitted TAT data for 7020 troponin and 4368 creatine kinase-MB determinations. On average, they reported 90% of myocardial injury marker results in slightly more than 90 minutes measured from the time that those tests were ordered. Among the fastest performing 25% of participants (75th percentile and above), median order-to-report troponin and creatine kinase-MB TATs were equal to 50 and 48.3 minutes or less, respectively. Shorter troponin TATs were associated with performing cardiac marker studies in EDs or other peripheral laboratories compared to (1) performing tests in central hospital laboratories, and (2) having cardiac marker specimens obtained by laboratory rather than by nonlaboratory personnel. CONCLUSION: The TAT expectations of the ED physicians using the results of laboratory tests measuring myocardial injury exceed those of the laboratory personnel providing the results of those tests. The actual TATs of myocardial injury testing meet the expectations of neither the providers of those tests nor the users of those test results. Improving TAT performance will require that the providers and users of laboratory services work together to develop standards that meet the needs of the medical staff and that are reasonably achievable by laboratory personnel. PMID- 14736290 TI - Detection of adenocarcinoma in situ of the cervix in Papanicolaou tests: comparison of diagnostic accuracy with other high-grade lesions. AB - CONTEXT: Adenocarcinoma in situ of the cervix is a recently recognized interpretation in the Bethesda 2001 system. Although specific morphologic criteria have been published, recognizing this entity is still difficult. OBJECTIVE: To compare pathologists' ability to correctly identify and categorize adenocarcinoma in situ with their ability to identify and categorize adenocarcinoma, high-grade squamous intraepithelial lesion, and squamous cell carcinoma. DESIGN: Pathologists' reviews in the 2001 and 2002 College of American Pathologists Interlaboratory Comparison Program in Cervicovaginal Cytology Program, an interlaboratory comparison program for gynecologic cytology, were examined. Cases were usually reviewed by multiple pathologists. False-negative rates, the percentage of reviews with exact agreement with reference interpretations, and the percentage of cases in which all reviews were in exact agreement with the reference interpretation for adenocarcinoma in situ, adenocarcinoma, high-grade squamous intraepithelial lesion, and squamous cell carcinoma were compared. RESULTS: A total of 213 reviews of cases categorized as adenocarcinoma in situ were compared with 2821 reviews of adenocarcinoma, 7535 reviews of high-grade squamous intraepithelial lesion, and 1886 reviews of squamous cell carcinoma. The false-negative rate for adenocarcinoma in situ (11.7%) was significantly higher than that for high-grade squamous intraepithelial lesion (4.6%, P <.001) and squamous cell carcinoma (3.3%, P <.001) but not for adenocarcinoma (8.9%, P =.16). Of all the reviews of adenocarcinoma in situ cases, 46.5% were interpreted specifically as adenocarcinoma in situ, compared to 72.2% of reviews of adenocarcinoma, 73.2% of high-grade squamous intraepithelial lesion, and 75.1% of squamous cell carcinoma. No individual case of adenocarcinoma in situ was always specifically recognized as adenocarcinoma in situ; 26.5% of cases of adenocarcinoma were specifically recognized as such in all reviews. Findings were similar with and without the inclusion of high-grade squamous intraepithelial lesion/carcinoma, not otherwise specified, as an acceptable review interpretation for cases of adenocarcinoma, squamous cell carcinoma, and high-grade squamous intraepithelial lesion. CONCLUSION: These data from expert-referenced and biopsy-proven cases suggest that adenocarcinoma in situ is not as easily recognized or categorized as other serious diagnoses. PMID- 14736292 TI - Multiple causes of difficulty in insight: the case of the nine-dot problem. AB - Theories of insight problems are often tested by formulating hypotheses about the particular difficulties of individual insight problems. Such evaluations often implicitly assume that there is a single difficulty. We argue that the quantitatively small effects of many studies arise because the difficulty of many insight problems is determined by multiple factors, so the removal of 1 factor has limited effect on the solution rate. Difficulties can reside either in problem perception, in prior knowledge, or in the processing of the problem information. We support this multiple factors perspective through 3 experiments on the 9-dot problem (N.R.F. Maier, 1930). Our results lead to a significant reformulation of the classical hypothesis as to why this problem is difficult. The results have general implications for our understanding of insight problem solving and for the interpretation of data from studies that aim to evaluate hypotheses about the sources of difficulty of particular insight problems. PMID- 14736293 TI - What makes an insight problem? The roles of heuristics, goal conception, and solution recoding in knowledge-lean problems. AB - Four experiments investigated transformation problems with insight characteristics. In Experiment 1, performance on a version of the 6-coin problem that had a concrete and visualizable solution followed a hill-climbing heuristic. Experiment 2 demonstrated that the difficulty of a version of the problem that potentially required insight for solution stems from the same hill-climbing heuristic, which creates an implicit conceptual block. Experiment 3 confirmed that the difficulty of the potential insight solution is conceptual, not procedural. Experiment 4 demonstrated the same principles of move selection on the 6-coin problem and the 10-coin (triangle) problem. It is argued that hill climbing heuristics provide a common framework for understanding transformation and insight problem solving. Postsolution receding may account for part of the phenomenology of insight. PMID- 14736294 TI - Consequential conditionals: invited and suppressed inferences from valued outcomes. AB - Consequential conditionals are defined as "if P then Q" statements, where P is an action, and Q a predicted outcome of this action, which is either desirable or undesirable to the agent. Experiment 1 shows that desirable (viz. undesirable) outcomes invite an inference to the truth (viz. falsity) of their antecedent. Experiment 2 shows that the more extreme the outcome is, the stronger the invited inference is. Experiment 3 shows that modus ponens from premises "If A then C, A" can be suppressed with the introduction of a consequential conditional, "If C then Q," where Q is an undesirable outcome. Experiment 4 shows that the more undesirable Q is, the larger the suppression is. The authors discuss how these results can enrich current approaches of conditional inference on the basis of mental models, complementary necessary conditions, and conditional probabilities. PMID- 14736295 TI - Nonmonotonic extrapolation in function learning. AB - This article reports the results of an experiment addressing extrapolation in function learning, in particular the issue of whether participants can extrapolate in a nonmonotonic manner. Existing models of function learning, including the extrapolation association model of function learning (EXAM; E. L. DeLosh, J. R. Busemeyer, & M. A. McDaniel, 1997), cannot account for this type of extrapolation pattern. We present the results of an experiment in which participants were shown a series of paired stimulus-response magnitudes where the relationship between these 2 dimensions conformed to a cyclic function. Participants were shown to extrapolate from these training data in a nonmonotonic way, contrary to predictions from EXAM. A new model of function learning is presented, which predicts responses more accurately than EXAM. PMID- 14736296 TI - Interactive effects of numerical surface form and operand parity in cognitive arithmetic. AB - In Experiment 1, adults (n = 48) performed simple addition, multiplication, and parity (i.e., odd-even) comparisons on pairs of Arabic digits or English number words. For addition and comparison, but not multiplication, response time increased with the number of odd operands. For addition, but not comparison, this parity effect was greater for words than for digits. In Experiment 2, adults (n = 50) solved simple addition problems in digit and word format and reported their strategies (i.e., retrieval or procedures). Procedural strategies were used more for odd than even addends and much more for word than digit problems. The results indicate that problem encoding and answer retrieval processes for cognitive arithmetic are interactive rather than strictly additive stages. PMID- 14736298 TI - Modeling associative recognition: a comparison of two-high-threshold, two-high threshold signal detection, and mixture distribution models. AB - A 2-high-threshold signal detection (HTSDT) model, a mixture distribution (SON) model, and 2-high-threshold (HT) models with responses distributed over 1 or several response categories were fit to results of 6 experiments from 2 studies on associative recognition: R. Kelley and J. T. Wixted (2001) and A. P. Yonelinas (1997). HTSDT assumes that associative recognition is based on conscious recollection and familiarity assessment, whereas according to SON and HT, associative information results in a shift of familiarity. The modeling results cast doubt on the prominent role of conscious recollection, and as far as models are valid, parameter estimation suggests 2 processes in associative recognition: a shift in familiarity that is due to associative information and the determination of the source of familiarity of pairs. PMID- 14736297 TI - Strategy execution in cognitive skill learning: an item-level test of candidate models. AB - This article investigates the transition to memory-based performance that commonly occurs with practice on tasks that initially require use of a multistep algorithm. In an alphabet arithmetic task, item response times exhibited pronounced step-function decreases after moderate practice that were uniquely predicted by T. C. Rickard's (1997) component power laws model. The results challenge parallel strategy execution models as developed to date and they demonstrate that the shift to retrieval is an item-specific, as opposed to task general, learning phenomenon. The results also call into question the entire class of smooth speed-up functions as global empirical learning laws. It is shown that overlaying of averaged item fits on averaged data can provide a sensitive test for model sufficiency. Strategy probes agreed with strategy inferences that were based on step-function speed-up patterns, supporting the validity of the probing technique. PMID- 14736299 TI - Abolishing the word-length effect. AB - The authors report 2 experiments that compare the recall of long and short words in pure and mixed lists. In pure lists, long words were much more poorly remembered than short words. In mixed lists, this word-length effect was abolished and both the long and short words were recalled as well as short words in pure lists. These findings contradict current models that seek to explain the word-length effect in terms of item-based effects such as difficulty in assembling items, or in terms of list-based accounts of rehearsal speed. An alternative explanation, drawing on ideas of item complexity and item distinctiveness, is proposed. PMID- 14736300 TI - The role of context in producing item interactions and false memories. AB - Cued recall with an extralist cue poses a challenge for contemporary memory theory in that there is a need to explain how episodic and semantic information are combined. A parallel activation and intersection approach proposes one such means by assuming that an experimental cue will elicit its preexisting semantic network and a context cue will elicit a list memory. These 2 sources of information are then combined by focusing on information that is common to the 2 sources. Two key predictions of that approach are examined: (a) Combining semantic and episodic information can lead to item interactions and false memories, and (b) these effects are limited to memory tasks that involve an episodic context cue. Five experiments demonstrate such item interactions and false memories in cued recall but not in free association. Links are drawn between the use of context in this setting and in other settings. PMID- 14736301 TI - Using recall to reduce false recognition: diagnostic and disqualifying monitoring. AB - Whether recall of studied words (e.g., parsley, rosemary, thyme) could reduce false recognition of related lures (e.g., basil) was investigated. Subjects studied words from several categories for a final recognition memory test. Half of the subjects were given standard test instructions, and half were instructed to use recall to reduce false recognition. Manipulation checks indicated that the latter instructions did elicit a recall-to-reject strategy. However, false recognition was selectively reduced only when all the words from a category could be recalled (Experiment 1). When longer categories were used, thereby minimizing exhaustive recall, a recall-to-reject strategy was ineffective at reducing false recognition (Experiment 2). It is suggested that exhaustively recalling a category allowed subjects to disqualify the lure as having occurred, analogous to recall-to-reject demonstrations in other tasks. In contrast, partially recalling a category did not help to diagnose the lure as nonstudied. These findings constrain theories of recall-based monitoring processes. PMID- 14736302 TI - The acquisition of vector knowledge and its relation to self-rated direction sense. AB - Self-rated sense of direction is reliably related to people's accuracy when pointing in the direction of unseen landmarks from imagined or actual perspectives. It is proposed that the cognitive substrate of accurate pointing responses is a vector representation, which is defined as an integrated network of displacement vectors. Experiment 1 isolated the body senses and tested displacement vector formation in a path-integration task. Experiment 2 isolated the visual modality and tested displacement vector formation in a virtual learning task. Both experiments tested whether people reporting a good sense of direction (GSOD) were more likely to compute displacement vectors than people reporting a poor sense of direction (PSOD). The results showed that both GSOD and PSOD people computed displacement vectors in the path-integration task, but not in the virtual-learning task. When interlandmark relations were visually specified, GSOD people made more accurate pointing responses than PSOD people, adding to a growing body of cognitive correlates of self-rated direction sense. PMID- 14736303 TI - Allocentric and egocentric updating of spatial memories. AB - In 4 experiments, the authors investigated spatial updating in a familiar environment. Participants learned locations of objects in a room, walked to the center, and turned to appropriate facing directions before making judgments of relative direction (e.g., "Imagine you are standing at X and facing Y. Point to Z.") or egocentric pointing judgments (e.g., "You are facing Y. Point to Z."). Experiments manipulated the angular difference between the learning heading and the imagined heading and the angular difference between the actual heading and the imagined heading. Pointing performance was best when the imagined heading was parallel to the learning view, even when participants were facing in other directions, and when actual and imagined headings were the same. Room geometry did not affect these results. These findings indicated that spatial reference directions in memory were not updated during locomotion. PMID- 14736304 TI - Orientation and perspective dependence in route and survey learning. AB - Three experiments investigated the role of egocentric orientation in subsequent memory for layouts learned via route (ground-level) and survey (aerial or overview) perspectives. Participants learned virtual environments from text descriptions (Experiment 1) or visual presentation (Experiments 1-3). In all experiments, scene recognition for route and survey images revealed a cost for switching perspective from study to test. In addition, recognition performance was facilitated when the test view matched the observer's learned orientation but only for the same-perspective recognition test. Experiment 3 demonstrated orientation dependence in judgments of relative direction, with a strong emphasis on initial heading. Together, these results suggest that establishing a reference system for representing spatial information is dependent on specific characteristics of the learning situation. PMID- 14736305 TI - Frames of reference in spatial memories acquired from language. AB - Four experiments examined reference systems in spatial memories acquired from language. Participants read narratives that located 4 objects in canonical (front, back, left, right) or noncanonical (left front, right front, left back, right back) positions around them. Participants' focus of attention was first set on each of the 4 objects, and then they were asked to report the name of the object at the location indicated by a direction word or an iconic arrow. The results indicated that spatial memories were represented in terms of intrinsic (object-to-object) reference systems, which were selected using egocentric cues (e.g., alignment with body axes). Results also indicated that linguistic direction cues were comprehended in terms of egocentric reference systems, whereas iconic arrows were not. PMID- 14736306 TI - Isolating the special component of face recognition: peripheral identification and a Mooney face. AB - A previous finding argues that, for faces, configural (holistic) processing can operate even in the complete absence of part-based contributions to recognition. Here, this result is confirmed using 2 methods. In both, recognition of inverted faces (parts only) was removed altogether (chance identification of faces in the periphery; no perception of a particularly hard-to-see Mooney face). Recognition of upright faces (configural plus parts), however, remained good. The simplicity of these new "isolation" techniques makes them ideal for (a) assessing configural processing in specialist populations (e.g., children, object experts) and (b) exploring properties of configural processing for faces in detail. As an example of the latter, orientation tuning was tested. Results argue against models in which faces are rotated to upright prior to identification. PMID- 14736307 TI - The dud-alternative effect in likelihood judgment. AB - The judged likelihood of a focal outcome should generally decrease as the list of alternative possibilities increases. For example, the likelihood that a runner will win a race goes down when 2 new entries are added to the field. However, 6 experiments demonstrate that the presence of implausible alternatives (duds) often increases the judged likelihood of a focal outcome. This dud-alternative effect was detected for judgments involving uncertainty about trivia facts and stochastic events. Nonnumeric likelihood measures and betting measures reliably detected the effect, but numeric likelihood measures did not. Time pressure increased the magnitude of the effect. The results were consistent with a contrast-effect account: The inclusion of duds increases the perceived strength of the evidence for the focal outcome, thereby affecting its judged likelihood. PMID- 14736308 TI - Diagnosticity and prototypicality in category learning: a comparison of inference learning and classification learning. AB - Category knowledge allows for both the determination of category membership and an understanding of what the members of a category are like. Diagnostic information is used to determine category membership; prototypical information reflects the most likely features given category membership. Two experiments examined 2 means of category learning, classification and inference learning, in terms of sensitivity to diagnostic and prototypical information. Classification learners were highly sensitive to diagnostic features but not sensitive to nondiagnostic, but prototypical, features. Inference learners were less sensitive to the diagnostic features than were classification learners and were also sensitive to the nondiagnostic, prototypical, features. Discussion focuses on aspects of the 2 learning tasks that might lead to this differential sensitivity and the implications for learning real-world categories. PMID- 14736309 TI - Category number impacts rule-based but not information-integration category learning: further evidence for dissociable category-learning systems. AB - Category number effects on rule-based and information-integration category learning were investigated. Category number affected accuracy and the distribution of best-fitting models in the rule-based task but had no effect on accuracy and little effect on the distribution of best-fining models in the information-integration task. In the 2 category conditions, rule-based learning was better than information-integration learning, whereas in the 4 category conditions, unidimensional and conjunctive rule-based learning was worse than information-integration learning. Rule-based strategies were used in the 2 category/rule-based condition, but about half of the observers used rule-based strategies in the 4-category unidimensional and conjunctive rule-based conditions. Information-integration strategies were used in the 4-category/ information-integration condition and by the end of training were used in the 2 category/information-integration condition. PMID- 14736310 TI - What is a TOT? Cognate and translation effects on tip-of-the-tongue states in Spanish-English and tagalog-English bilinguals. AB - The authors induced tip-of-the-tongue states (TOTs) for English words in monolinguals and bilinguals using picture stimuli with cognate (e.g., vampire, which is vampiro in Spanish) and noncognate (e.g., funnel, which is embudo in Spanish) names. Bilinguals had more TOTs than did monolinguals unless the target pictures had translatable cognate names, and bilinguals had fewer TOTs for noncognates they were later able to translate. TOT rates for the same targets in monolinguals indicated that these effects could not be attributed to target difficulty. Two popular TOT accounts must be modified to explain cognate and translatability facilitation effects, and cross-language interference cannot explain bilinguals' increased TOTs rates. Instead the authors propose that, relative to monolinguals, bilinguals are less able to activate representations specific to each language. PMID- 14736311 TI - Nonword repetition priming in lexical decision reverses as a function of study task and speed stress. AB - The authors argue that nonword repetition priming in lexical decision is the net result of 2 opposing processes. First, repeating nonwords in the lexical decision task results in the storage of a memory trace containing the interpretation that the letter string is a nonword; retrieval of this trace leads to an increase in performance for repeated nonwords. Second, nonword repetition results in increased familiarity, making the nonword more "wordlike," leading to a decrease in performance. Consistent with this dual-process account, Experiment 1 showed a facilitatory effect for nonwords studied in a lexical decision task but an inhibitory effect for nonwords studied in a letter-height task. Experiment 2 showed inhibitory nonword repetition priming for participants tested under speed stress instructions. PMID- 14736312 TI - What determines the speed of lexical access: homophone or specific-word frequency? A reply to Jescheniak et al. (2003). AB - A. Caramazza, A. Costa, M. Miozzo, and Y. Bi (2001) reported a series of experiments showing that naming latencies for homophones are determined by specific-word frequency (e.g., frequency of nun) and not homophone frequency (frequency of nun + none). J. D. Jescheniak, A. S. Meyer, and W. J. M. Levelt (2003) have challenged these studies on a variety of grounds. Here we argue that these criticisms are not well founded and try to clarify the theoretical issues that can be meaningfully addressed by considering the effects of frequency on homophone production. We conclude that the evidence from homophone production cannot be considered to provide support to 2-layer theories of the lexical system. PMID- 14736313 TI - Updating situation models. AB - The authors examined how situation models are updated during text comprehension. If comprehenders keep track of the evolving situation, they should update their models such that the most current information, the here and now, is more available than outdated information. Contrary to this updating hypothesis, E. J. O'Brien, M. L. Rizzella, J. E. Albrecht, and J. G. Halleran (1998) obtained results suggesting that outdated or incorrect information may still influence the comprehension process. The authors of the current study demonstrate that the nature of E. J. O'Brien et al.'s materials were the likely cause of this pattern of results. Hence, the current authors constructed materials that circumvent identified confounds and in a reading-time experiment obtained evidence supporting the here-and-now hypothesis. PMID- 14736315 TI - On interpreting stereotype threat as accounting for African American-White differences on cognitive tests. AB - C. M. Steele and J. Aronson (1995) showed that making race salient when taking a difficult test affected the performance of high-ability African American students, a phenomenon they termed stereotype threat. The authors document that this research is widely misinterpreted in both popular and scholarly publications as showing that eliminating stereotype threat eliminates the African American White difference in test performance. In fact, scores were statistically adjusted for differences in students' prior SAT performance, and thus, Steele and Aronson's findings actually showed that absent stereotype threat, the two groups differ to the degree that would be expected based on differences in prior SAT scores. The authors caution against interpreting the Steele and Aronson experiment as evidence that stereotype threat is the primary cause of African American-White differences in test performance. PMID- 14736316 TI - Should there be a three-strikes rule against pure discovery learning? The case for guided methods of instruction. AB - The author's thesis is that there is sufficient research evidence to make any reasonable person skeptical about the benefits of discovery learning--practiced under the guise of cognitive constructivism or social constructivism--as a preferred instructional method. The author reviews research on discovery of problem-solving rules culminating in the 1960s, discovery of conservation strategies culminating in the 1970s, and discovery of LOGO programming strategies culminating in the 1980s. In each case, guided discovery was more effective than pure discovery in helping students learn and transfer. Overall, the constructivist view of learning may be best supported by methods of instruction that involve cognitive activity rather than behavioral activity, instructional guidance rather than pure discovery, and curricular focus rather than unstructured exploration. PMID- 14736317 TI - Loss, trauma, and human resilience: have we underestimated the human capacity to thrive after extremely aversive events? AB - Many people are exposed to loss or potentially traumatic events at some point in their lives, and yet they continue to have positive emotional experiences and show only minor and transient disruptions in their ability to function. Unfortunately, because much of psychology's knowledge about how adults cope with loss or trauma has come from individuals who sought treatment or exhibited great distress, loss and trauma theorists have often viewed this type of resilience as either rare or pathological. The author challenges these assumptions by reviewing evidence that resilience represents a distinct trajectory from the process of recovery, that resilience in the face of loss or potential trauma is more common than is often believed, and that there are multiple and sometimes unexpected pathways to resilience. PMID- 14736318 TI - How the mind hurts and heals the body. AB - The author reviews some of the social and behavioral factors acting on the brain that influence health, illness, and death. Supported with data from several areas of research, his proposal for understanding health and illness provides both the concepts and the mechanisms for studying and explaining mind-body relationships. The brain is the body's first line of defense against illness, and the mind is the emergent functioning of the brain. This mind-body approach incorporates ideas, belief systems, and hopes as well as biochemistry, physiology, and anatomy. Changing thoughts imply a changing brain and thus a changing biology and body. Belief systems provide a baseline for the functioning brain upon which other variables act and have their effects. PMID- 14736319 TI - Theodore Hertzl Blau (1928-2003). PMID- 14736320 TI - John Bissell Carroll (1916-2003). PMID- 14736321 TI - Nancy Collier Waugh (1930-2002). PMID- 14736322 TI - Robert W. Swezey (1943-2002). PMID- 14736323 TI - Stereotype threat does not live by Steele and Aronson (1995) alone. PMID- 14736325 TI - Journal impact factors and self-citations: implications for psychology journals. PMID- 14736326 TI - On (not) trimming one's toenails with a bazooka. PMID- 14736327 TI - Galton on spirituality, religion, and health. PMID- 14736328 TI - Religion, spirituality, and health: a topic not so new. PMID- 14736329 TI - Religion, spirituality, and health. PMID- 14736330 TI - Unaddressed problems in the study of spirituality and health. PMID- 14736332 TI - Proceedings and abstracts from the NETwork/European Neuroendocrine Tumor Group meeting on neuroendocrine tumors. May 8-9, 2003. London, United Kingdom. PMID- 14736335 TI - Do pediatric hospitalizations have a unique geography? AB - BACKGROUND: In the U.S. small-area health services research studies are often based on the hospital service areas (HSAs) defined by the Dartmouth Atlas of Healthcare project. These areas are based on the geographic origins of Medicare Part A hospital patients, the great majority of whom are seniors. It is reasonable to question whether the geographic system so defined is appropriate for health services research for all ages, particularly for children, who have a very different system of healthcare financing and provision in the U.S. METHODS: This article assesses the need for a unique system of HSAs to support pediatric small-area analyses. It is a cross-sectional analysis of California hospital discharges for two age groups - non-newborns 0-17 years old, and seniors. The measure of interest was index of localization, which is the percentage of HSA residents hospitalized in their home HSA. Indices were computed separately for each age group, and index agreement was assessed for 219 of the state's HSAs. We examined the effect of local pediatric inpatient volume and pediatric inpatient resources on the divergence of the age group indices. We also created a new system of HSAs based solely on pediatric patient origins, and visually compared maps of the traditional and the new system. RESULTS: The mean localization index for pediatric discharges was 20 percentage points lower than for Medicare cases, indicating a poorer fit of the traditional geographic system for children. The volume of pediatric cases did not appear to be associated with the magnitude of index divergence between the two age groups. Pediatric medical and surgical case subgroups gave very similar results, and both groups differed substantially from seniors. Location of children's hospitals and local pediatric bed supply were associated with Medicare-pediatric divergence. There was little visual correspondence between the maps of traditional and pediatric-specific HSAs. CONCLUSION: Children and seniors have significantly different geographic patterns of hospitalization in California. Medicare-based HSAs may not be appropriate for all age groups and service types throughout the U.S. PMID- 14736333 TI - Rhesus monkey gastric mucins: oligomeric structure, glycoforms and Helicobacter pylori binding. AB - Mucins isolated from the stomach of Rhesus monkey are oligomeric glycoproteins with a similar mass, density, glycoform profile and tissue localization as human MUC5AC and MUC6. Antibodies raised against the human mucins recognize those from monkey, which thus appear to be orthologous to those from human beings. Rhesus monkey muc5ac and muc6 are produced by the gastric-surface epithelium and glands respectively, and occur as three distinct glycoforms. The mucins are substituted with the histo blood-group antigens B, Le(a) (Lewis a), Le(b), Le(x), Le(y), H type-2, the Tn-antigen, the T-antigen, the sialyl-Le(x) and sialyl-Le(a) structures, and the expression of these determinants varies between individuals. At neutral pH, Helicobacter pylori strains expressing BabA (blood-group antigen binding adhesin) bind Rhesus monkey gastric mucins via the Le(b) or H-type-1 structures, apparently on muc5ac, as well as on a smaller putative mucin, and binding is inhibited by Le(b) or H-type-1 conjugates. A SabA (sialic acid-binding adhesin)-positive H. pylori mutant binds to sialyl-Le(x)-positive mucins to a smaller extent compared with the BabA-positive strains. At acidic pH, the microbe binds to mucins substituted by sialylated structures such as sialyl-Le(x) and sialylated type-2 core, and this binding is inhibited by DNA and dextran sulphate. Thus mucin- H. pylori binding occurs via at least three different mechanisms: (1) BabA-dependent binding to Le(b) and related structures, (2) SabA dependent binding to sialyl-Le(x) and (3) binding through a charge-mediated mechanism to sialylated structures at low pH values. PMID- 14736336 TI - Unintended pregnancy and induced abortion among unmarried women in China: a systematic review. AB - BACKGROUND: Until recently, premarital examination for both men and women was a legal requirement before marriage in China. Researchers have carried out surveys of attendees' sexual activity, pregnancy and abortion before their marriages, trying to map out reproductive health needs in China, according to this unique population-based data. To systematically identify, appraise and summarise all available studies documenting pregnancy and induced abortion among unmarried Chinese women attending premarital examinations. METHODS: We searched the Chinese Biomedical Literature Index from 1978 to 2002; PUBMED; and EMBASE. Trials were assessed and data extracted by two people independently. RESULTS: Nine studies, of which seven were conducted in the urban areas, one in the rural areas, and one in both urban and rural areas, met the inclusion criteria. In the seven studies in urban areas, the majority of unmarried women had experienced sexual intercourse, with estimates ranging from 54% to 82% in five studies. Estimates of a previous pregnancy ranged from 12% to 32%. Abortion rates were high, ranging between 11 to 55% in 8 studies reporting this, which exclude the one rural study. In the three studies reporting both pregnancy and abortion, most women who had become pregnant had an induced abortion (range 86% to 96%). One large rural study documented a lower low pregnancy rate (20%) and induced abortion rate (0.8%). CONCLUSIONS: There is a large unmet need for temporary methods of contraception in urban areas of China. PMID- 14736337 TI - Sentinel node biopsy for breast cancer: is it already a standard of care? A survey of current practice in an Italian region. AB - BACKGROUND: Although sentinel node biopsy (SNB) is becoming the standard approach for axillary staging in patients with small breast cancer, criteria for patient selection and some technical aspects of the procedure have yet to be clearly defined. The aim of the present survey was therefore to investigate the way in which SNB is used by general surgeons working in the Veneto region, Italy. METHODS: A 29-item questionnaire regarding various aspects of SNB practice was mailed to surgeons in charge of breast surgery in all the 56 surgical centres of the region. RESULTS: The rate of response to the questionnaire was 82.1% (n = 46); 69.6% (n = 32) of the respondents routinely perform SNB in their clinical practice. Most of the interviewed surgeons (93.5%) expressed the belief that the acceptable false negative rate should be < or =5%. However, among the surgeons who perform SNB, only 34.4% performed more than 20 SNB during the learning phase. Indications are limited to tumours of < or =1 cm by 31.2% (n = 10) of respondents, < or =2 cm by 46.9% (n = 15) and < or =3 cm by 21.9% (n = 7). Almost all respondents (93.7%) agreed that a clinically positive axilla is a contraindication to SNB, while opinions differed widely concerning other potential contraindications. In most of the centres considered, SN identification is undertaken on the day before surgery using a subdermal injection of 30-50 MBq of 99mTc-albumin-nanocolloid followed by lymphoscintigraphy. CONCLUSIONS: SNB is currently performed in the majority of hospitals in the Veneto region. However, the training phase and criteria used for patient selection differ from centre to centre. Certified training courses and shared guidelines are therefore highly desirable. PMID- 14736338 TI - Sam68 exerts separable effects on cell cycle progression and apoptosis. AB - BACKGROUND: The RNA-binding protein Sam68 has been implicated in a number of cellular processes, including transcription, RNA splicing and export, translation, signal transduction, cell cycle progression and replication of the human immunodeficiency virus and poliovirus. However, the precise impact it has on essential cellular functions remains largely obscure. RESULTS: In this report we show that conditional overexpression of Sam68 in fibroblasts results in both cell cycle arrest and apoptosis. Arrest in G1 phase of the cell cycle is associated with decreased levels of cyclins D1 and E RNA and protein, resulting in dramatically reduced Rb phosphorylation. Interestingly, cell cycle arrest does not require the specific RNA binding ability of Sam68. In marked contrast, induction of apoptosis by Sam68 absolutely requires a fully-functional RNA binding domain. Moreover, the anti-cancer agent trichostatin A potentiates Sam68 driven apoptosis. CONCLUSIONS: For the first time we have shown that Sam68, an RNA binding protein with multiple apparent functions, exerts functionally separable effects on cell proliferation and survival, dependent on its ability to bind specifically to RNA. These findings shed new light on the ability of signal transducing RNA binding proteins to influence essential cell function. Moreover, the ability of a class of anti-cancer therapeutics to modulate its ability to promote apoptosis suggests that Sam68 status may impact some cancer treatments. PMID- 14736339 TI - Protein kinase Czeta regulates phospholipase D activity in rat-1 fibroblasts expressing the alpha1A adrenergic receptor. AB - BACKGROUND: Phenylephrine (PHE), an alpha1 adrenergic receptor agonist, increases phospholipase D (PLD) activity, independent of classical and novel protein kinase C (PKC) isoforms, in rat-1 fibroblasts expressing alpha1A adrenergic receptors. The aim of this study was to determine the contribution of atypical PKCzeta to PLD activation in response to PHE in these cells. RESULTS: PHE stimulated a PLD activity as demonstrated by phosphatidylethanol production. PHE increased PKCzeta translocation to the particulate cell fraction in parallel with a time-dependent decrease in its activity. PKCzeta activity was reduced at 2 and 5 min and returned to a sub-basal level within 10-15 min. Ectopic expression of kinase-dead PKCzeta, but not constitutively active PKCzeta, potentiated PLD activation elicited by PHE. A cell-permeable pseudosubstrate inhibitor of PKCzeta reduced basal PKCzeta activity and abolished PHE-induced PLD activation. CONCLUSION: alpha1A adrenergic receptor stimulation promotes the activation of a PLD activity by a mechanism dependent on PKCzeta; Our data also suggest that catalytic activation of PKCzeta is not required for PLD stimulation. PMID- 14736340 TI - HMM Logos for visualization of protein families. AB - BACKGROUND: Profile Hidden Markov Models (pHMMs) are a widely used tool for protein family research. Up to now, however, there exists no method to visualize all of their central aspects graphically in an intuitively understandable way. RESULTS: We present a visualization method that incorporates both emission and transition probabilities of the pHMM, thus extending sequence logos introduced by Schneider and Stephens. For each emitting state of the pHMM, we display a stack of letters. The stack height is determined by the deviation of the position's letter emission frequencies from the background frequencies. The stack width visualizes both the probability of reaching the state (the hitting probability) and the expected number of letters the state emits during a pass through the model (the state's expected contribution).A web interface offering online creation of HMM Logos and the corresponding source code can be found at the Logos web server of the Max Planck Institute for Molecular Genetics http://logos.molgen.mpg.de. CONCLUSIONS: We demonstrate that HMM Logos can be a useful tool for the biologist: We use them to highlight differences between two homologous subfamilies of GTPases, Rab and Ras, and we show that they are able to indicate structural elements of Ras. PMID- 14736341 TI - Benchmarking tools for the alignment of functional noncoding DNA. AB - BACKGROUND: Numerous tools have been developed to align genomic sequences. However, their relative performance in specific applications remains poorly characterized. Alignments of protein-coding sequences typically have been benchmarked against "correct" alignments inferred from structural data. For noncoding sequences, where such independent validation is lacking, simulation provides an effective means to generate "correct" alignments with which to benchmark alignment tools. RESULTS: Using rates of noncoding sequence evolution estimated from the genus Drosophila, we simulated alignments over a range of divergence times under varying models incorporating point substitution, insertion/deletion events, and short blocks of constrained sequences such as those found in cis-regulatory regions. We then compared "correct" alignments generated by a modified version of the ROSE simulation platform to alignments of the simulated derived sequences produced by eight pairwise alignment tools (Avid, BlastZ, Chaos, ClustalW, DiAlign, Lagan, Needle, and WABA) to determine the off the-shelf performance of each tool. As expected, the ability to align noncoding sequences accurately decreases with increasing divergence for all tools, and declines faster in the presence of insertion/deletion evolution. Global alignment tools (Avid, ClustalW, Lagan, and Needle) typically have higher sensitivity over entire noncoding sequences as well as in constrained sequences. Local tools (BlastZ, Chaos, and WABA) have lower overall sensitivity as a consequence of incomplete coverage, but have high specificity to detect constrained sequences as well as high sensitivity within the subset of sequences they align. Tools such as DiAlign, which generate both local and global outputs, produce alignments of constrained sequences with both high sensitivity and specificity for divergence distances in the range of 1.25-3.0 substitutions per site. CONCLUSION: For species with genomic properties similar to Drosophila, we conclude that a single pair of optimally diverged species analyzed with a high performance alignment tool can yield accurate and specific alignments of functionally constrained noncoding sequences. Further algorithm development, optimization of alignment parameters, and benchmarking studies will be necessary to extract the maximal biological information from alignments of functional noncoding DNA. PMID- 14736342 TI - Prediction of falls using a risk assessment tool in the acute care setting. AB - BACKGROUND: The British STRATIFY tool was previously developed to predict falls in hospital. Although the tool has several strengths, certain limitations exist which may not allow generalizability to a Canadian setting. Thus, we tested the STRATIFY tool with some modification and re-weighting of items in Canadian hospitals. METHODS: This was a prospective validation cohort study in four acute care medical units of two teaching hospitals in Hamilton, Ontario. In total, 620 patients over the age of 65 years admitted during a 6-month period. Five patient characteristics found to be risk factors for falls in the British STRATIFY study were tested for predictive validity. The characteristics included history of falls, mental impairment, visual impairment, toileting, and dependency in transfers and mobility. Multivariate logistic regression was used to obtain optimal weights for the construction of a risk score. A receiver-operating characteristic curve was generated to show sensitivities and specificities for predicting falls based on different threshold scores for considering patients at high risk. RESULTS: Inter-rater reliability for the weighted risk score indicated very good agreement (inter-class correlation coefficient = 0.78). History of falls, mental impairment, toileting difficulties, and dependency in transfer / mobility significantly predicted fallers. In the multivariate model, mental status was a significant predictor (P < 0.001) while history of falls and transfer / mobility difficulties approached significance (P = 0.089 and P = 0.077 respectively). The logistic regression model led to weights for a risk score on a 30-point scale. A risk score of 9 or more gave a sensitivity of 91% and specificity of 60% for predicting who would fall. CONCLUSION: Good predictive validity for identifying fallers was achieved in a Canadian setting using a simple-to-obtain risk score that can easily be incorporated into practice. PMID- 14736343 TI - Do younger women with non-metastatic and non-inflammatory breast carcinoma have poor prognosis? AB - BACKGROUND: Controversy abounds over whether breast cancer in younger women is more aggressive than those in older. The aim of the study was to assess the influence of age on long-term survival of women with breast carcinoma. MATERIALS AND METHODS: Patients with non-metastatic and non-inflammatory invasive breast carcinoma treated at the Regional Cancer Centre, Trivandrum, Kerala, India during 1990-93 were divided into 4 age groups as < 40 years, 40-49, 50-59, and > 60 years. The overall survival (OS) for each age group was estimated using the Kaplan-Meier method in relation to the primary tumor (T) and the axillary node status (N). The OS of the various age groups were compared using the log-rank test. Hazard ratio and 95% confidence interval for each age group was estimated using Cox-regression model after adjusting for T and N. RESULTS: Between 1990 1993, 1701 women (26%, <40 years) reported with non-metastatic and non inflammatory invasive breast carcinoma. Overall survival (OS) of all the women was 52.6 % (standard error 1%) at 10 years. The OS for women with age < 40 years and with T3 and T4 disease status was 36.6% and 10.4% respectively and for those in 40-49 age group was 41.9% and 33.5%. The 10-year OS for women with node positive (N1) disease was 24.6% in < 40 years and 45.2% in the 40-49 age group (p = 0.0006). After adjusting for tumor and node stage the relative risk for death was 24% lower for women in 40-49 age group as compared to women <40 years of age. CONCLUSION: Women under 40 years with T3/ T4 breast lesions and/or positive axillary nodes were found to have a significantly poorer survival. PMID- 14736344 TI - Agreement between insurance claim and self-reported hospital and emergency room utilization data among persons with diabetes. AB - As part of a retrospective evaluation of a diabetes management program, the agreement between self-reported and insurance claim data on hospitalization and emergency room utilization was examined. Data agreement on hospitalization or emergency room visits between the two collection modes was evaluated through the use of simple agreement proportions and the kappa agreement statistic. A total of 1,230 participant responses were studied. The proportions of patients with hospitalization or emergency room visits were indistinguishable between the self reported and medical claims data, and kappa statistics also indicated good-to excellent agreement between data sets. The percentages of participants whose self reported hospitalization and emergency room utilization exactly matched data derived from insurance claims were high (89.1% and 87.2%, respectively). Furthermore, the kappa statistics of agreement for the number of hospitalizations (0.6366) and emergency room visits (0.5390) indicate good agreement between self reported and insurance claim data. The results of this study suggest either self reported or insurance claims data can be used to evaluate the impact of health care interventions on hospital or emergency room utilization. PMID- 14736345 TI - Cost savings for a preferred provider organization population with multi condition disease management: evaluating program impact using predictive modeling with a control group. AB - Disease management programs have historically had difficulty demonstrating the financial value of their programs using statistically rigorous methods. This preliminary study utilized a predictive model built from a control group that consisted of individual employer groups whose benefits managers did not purchase disease management. The purpose of this evaluation was to examine the cost savings associated with Health Management Corporation's disease management program for asthma, diabetes, and coronary artery disease. Study and control group members were identified with exactly the same selection criteria as self insured employer groups (n = 76,194) with preferred provider organization health plans that offered the disease management program. The study group consisted of members (n = 1,009) who were identified as having any of the three conditions and were eligible for the disease management program, regardless of the extent of their participation. The control group included members (n = 2,491) who were identified as having any of the same three conditions. The control group data were used to develop a predictive model designed to calculate expected medical claims costs for the study group. The gross savings of the disease management program was measured by calculating the difference between the expected medical claims costs predicted by the model and the actual medical claims costs for the study group. Preliminary analyses indicate that Health Management Corporation's disease management program produced a return on investment of $2.84: $1.00 and $1.45 gross savings per member per month. PMID- 14736346 TI - Emerging information management technologies and the future of disease management. AB - Disease management (DM) has become a widely accepted way to support care delivery in the chronically ill patient population. Patients enrolled in these programs have been shown to have better health, fewer complications and comorbidities, and lower health care costs. The development of advanced information management technologies is further enhancing the role DM plays in optimizing outcomes and cost-effectiveness in clinical care. These emerging information management technologies (EIMT) include advances in software, hardware, and networking, all of which share common impact attributes in their ability to improve cost effectiveness of care, quality of care, and access to care. Specific examples include interactive websites with the ability to engage patients in the self-care management process, the embedding of biometric devices (digital scales, modem enabled glucose meters in the home, blood pressure monitoring, etc.), workflow and care coordination programs that add intelligence via guideline-directed alerts and reminders to the delivery process, registries that include a summary of personal health data that can be used as a reference point for improved clinical decisions, and the systematic collection of aggregated, de-identified clinical, administrative, and cost data into comprehensive data sets to which predictive modeling analytic tools can be applied. By way of case example, we also present data from a controlled clinical trial utilizing EIMT in the form of home-based weight measurement using a digital scale and linkage to a care coordination center for the management of severe congestive heart failure. Outcome results on 85,515 patient-months of an aggregate commercial and Medicare continuously enrolled population demonstrated an average reduction of care utilization (hospitalization) of 57% and a reduction in related delivery cost (per member per year payments) of 55%. We conclude that EIMT have already begun to offer significant and quantifiable benefits to DM and are likely to become heavily embedded in care management strategies in the future. PMID- 14736347 TI - Quality improvement and cost reduction realized by a purchaser through diabetes disease management. AB - This report documents the clinical improvements and costs experienced by a purchaser after introduction of a diabetes disease management program. A purchaser contracted with American Healthways, a disease management organization, to initiate a diabetes disease management program called Diabetes Decisions. Started in 1998, the program grew to include 662 participants. The results reported are based on the continuously participating population (12 months of participation in the program for the reporting year). Participants were entered into American Healthways' clinical information system and risk-stratified, and an individualized treatment plan was devised. Outbound telephone calls by specially trained nurses were a key intervention. Data were collected on key process measures, financial parameters, and participant satisfaction. By year 3, there were 422 continuously participating participants. From baseline to the third year of the program, significant increases in frequency of A1C testing (21.3% to 82.2%), dilated retinal exams (17.2% to 70.7%), and performance of foot exams (2.0% to 75.6%) were noted. For 166 participants with five A1C determinations, A1C values dropped from 8.89% to 7.88%. Participants experienced a 36% drop in inpatient costs. Without adjustment for medical inflation, total medical costs decreased by 26.8% from the baseline period, dropping to $268.63 per diabetes participant per month (PDPPM) by year 3, a gross savings of $98.49 PDPPM. After subtracting the fees paid to Diabetes Decisions, a net savings of $986,538 was realized. This yielded a return on investment of 3.37. By investing in a diabetes disease management program, a purchaser was able to realize significant improvements in clinical care, substantial cost savings, and a favorable return on investment. PMID- 14736348 TI - Evaluating disease management program effectiveness: an introduction to time series analysis. AB - Currently, the most widely used method in the disease management (DM) industry for evaluating program effectiveness is referred to as the "total population approach." This model is a pretest-posttest design, with the most basic limitation being that without a control group, there may be sources of bias and/or competing extraneous confounding factors that offer a plausible rationale explaining the change from baseline. Furthermore, with the current inclination of DM programs to use financial indicators rather than program-specific utilization indicators as the principal measure of program success, additional biases are introduced that may cloud evaluation results. This paper presents a non-technical introduction to time-series analysis (using disease-specific utilization measures) as an alternative, and more appropriate, approach to evaluating DM program effectiveness than the current total population approach. PMID- 14736349 TI - A new partnership: the Society for Acupuncture Research and the Journal of Alternative and Complementary Medicine. PMID- 14736350 TI - Can CAM come in from the cold? An update on the ongoing regulatory process with regard to herbal and acupuncture practitioners in the United Kingdom. PMID- 14736351 TI - Importance of complementary and alternative cancer therapies in palliative oncology in India. PMID- 14736352 TI - Biocultural diversity, phytomedicines, and tropical rainforests: the holistic link from practitioner to cultures of the tropical rainforest. PMID- 14736353 TI - Improvements in muscle symmetry in children with cerebral palsy after equine assisted therapy (hippotherapy). AB - OBJECTIVE: To evaluate the effect of hippotherapy (physical therapy utilizing the movement of a horse) on muscle activity in children with spastic cerebral palsy. DESIGN: Pretest/post-test control group. SETTING/LOCATION: Therapeutic Riding of Tucson (TROT), Tucson, AZ. SUBJECTS: Fifteen (15) children ranging from 4 to 12 years of age diagnosed with spastic cerebral palsy. INTERVENTIONS: Children meeting inclusion criteria were randomized to either 8 minutes of hippotherapy or 8 minutes astride a stationary barrel. OUTCOME MEASURES: Remote surface electromyography (EMG) was used to measure muscle activity of the trunk and upper legs during sitting, standing, and walking tasks before and after each intervention. RESULTS: After hippotherapy, significant improvement in symmetry of muscle activity was noted in those muscle groups displaying the highest asymmetry prior to hippotherapy. No significant change was noted after sitting astride a barrel. CONCLUSIONS: Eight minutes of hippotherapy, but not stationary sitting astride a barrel, resulted in improved symmetry in muscle activity in children with spastic cerebral palsy. These results suggest that the movement of the horse rather than passive stretching accounts for the measured improvements. PMID- 14736354 TI - Benefits of external qigong therapy on morphine-abstinent mice and rats. AB - OBJECTIVE: To exclude possible psychological effects of qigong therapy in the treatment of addiction effectively, morphine-dependence models need to be established in mice and rats. METHOD: The effects of external qi on withdrawal syndrome were examined in naloxone-precipitated mice and rats in three randomized control experiments: naloxone-precipitated test in morphine-dependent mice (n = 100 in 5 groups, 20 mice each group); conditioned position preference test in morphine-abstinent mice (n = 30 for 3 groups, 10 each); and naloxone-precipitated test with paired box in morphine-dependent rats (n = 40 for 4 groups, 10 each). RESULTS: These experiments showed that morphine-dependent mice, after external qigong (EQ) therapy, had decreased incidence of jumping and lower jumping frequencies, and attenuated loss of body weight. After EQ therapy, morphine dependent rats had reduced withdrawal scores and body weight loss was inhibited. In the conditioned place preference test, the time spent in the drug-paired box was significantly shorter for the qigong group than for the morphine group. CONCLUSION: These results suggest that qigong might have an inhibitory effect on withdrawal syndrome, and reduce the dependence potential in mice. Three different designs confirm that the impact of qigong therapy on morphine-abstinent mice and rats is reliable and substantial. Further research on the effectiveness and the mechanism of qigong therapy on addiction is warranted. PMID- 14736355 TI - Efficacy of massage therapy in chronic pain: a pragmatic randomized trial. AB - BACKGROUND: Although classic massage is used widely in Germany and elsewhere for treating chronic pain conditions, there are no randomized controlled trials (RCT). DESIGN: Pragmatic RCT of classic massage compared to standard medical care (SMC) in chronic pain conditions of back, neck, shoulders, head and limbs. OUTCOME MEASURE: Pain rating (nine-point Likert-scale; predefined main outcome criterion) at pretreatment, post-treatment, and 3 month follow-up, as well as pain adjective list, depression, anxiety, mood, and body concept. RESULTS: Because of political and organizational problems, only 29 patients were randomized, 19 to receive massage, 10 to SMC. Pain improved significantly in both groups, but only in the massage group was it still significantly improved at follow-up. Depression and anxiety were improved significantly by both treatments, yet only in the massage group maintained at follow-up. CONCLUSION: Despite its limitation resulting from problems with numbers and randomization this study shows that massage can be at least as effective as SMC in chronic pain syndromes. Relative changes are equal, but tend to last longer and to generalize more into psychologic domains. Because this is a pilot study, the results need replication, but our experiences might be useful for other researchers. PMID- 14736356 TI - Single-needle acupuncture alleviates gag reflex during transesophageal echocardiography: a blinded, randomized, controlled pilot trial. AB - OBJECTIVES: To study the effect of single-needle acupuncture in suppressing gag reflex in transesophageal echocardiography (TEE). DESIGN: Prospective, blinded trial. Settings/locations: Patients with ischemic stroke or transient ischemic attack undergoing TEE because of presumed cardioembolic origin in a specialized stroke unit of the Johann Wolfgang Goethe-University, Frankfurt/Main, Germany. Subjects/Study interventions: Forty-one (41) patients were studied. Patients received single-needle acupuncture with a 0.2 x 13 mm disposable acupuncture needle (Suzhou Medical Appliances, China), 10-mm deep either at Chengjiang (midline between lower lip and chin) or superficially at a sham point (tip of the chin) during TEE or no acupuncture for alleviating gag reflex. OUTCOME MEASURES: Severity of gagging was rated on a visual-analogue scale. RESULTS: The acupuncture group experienced significantly less gagging than the sham group (p = 0.037) or the nonacupuncture group (p = 0.013). CONCLUSIONS: Acupuncture of CV24 is an easy to apply and effective method to reduce gag reflex during TEE. PMID- 14736357 TI - Perivascular space: possible anatomical substrate for the meridian. AB - BACKGROUND: Despite the meridian system being an important concept in Traditional Chinese Medicine (TCM), modern biology and Western medical systems have failed to find an anatomic substrate. Since the 1960s, a variety of phenomena along meridians have been reported, among which quite a few suggest that along meridians there is a fluid pathway (but not blood vessels or lymphatics). On the other hand, perivascular space (PVS) has been demonstrated to be a body fluid pathway in addition to blood vessels and lymphatics in some mammalian tissues, such as brain, thymus, and lung. OBJECTIVES: The present study was designed to examine the relationship between PVS and the meridian. We studied characteristics of the tissues around the blood vessels along the Stomach Meridian of Foot Yangming and the Gallbladder Meridian of Foot-shaoyang, with the goal of identifying anatomical structure corresponding to the meridian described in TCM. DESIGN AND RESULTS: Through perivascular dye injection and frozen section histology, we found that there is PVS around the blood vessels along the meridians, and it is a fluid pathway. Subsequent physiologic studies revealed that the PVS shows significantly greater electrical conductivity and significantly higher partial oxygen pressure (pO(2)) compared to medial and lateral tissues. CONCLUSIONS: The PVS along the meridians has properties offering good explanation for the meridian phenomena. The work sheds new light on the studies of meridians and may contribute to research on the mechanism of Chinese acupuncture. PMID- 14736358 TI - Alternative method of gifted identification using the AMI: an apparatus for measuring internal meridians and their corresponding organs. AB - OBJECTIVES: To determine if meridian energy (qi or bioelectricity), as measured by the Apparatus for Measuring the Internal Organs and their Corresponding Meridians (AMI), could distinguish between gifted and average boys and to gather evidence to assess if the AMI device could be effective as an educational diagnostic tool, specifically for identifying gifted children. DESIGN: Quasi experimental design using discriminant analysis to determine how accurately the AMI could predict group membership previously determined by intelligence tests. SETTINGS/LOCATION: Practitioner's office within clinic in Albuquerque, New Mexico. SUBJECTS: Thirty-three (33 boys; gifted, n = 11 and average, n = 22) ranging in age from 9 to 12 years. OUTCOME MEASURES: Meridian nervous system energy levels as measured by the AMI. RESULTS: On average, boys in the gifted group had statistically significantly calmer nervous system measures than the boys in the average group. The discriminant function generated from the AMI nervous system measures accurately predicted group membership 70% of the time. Classification accuracy by groups indicated that 55% of the gifted cases and 82% of the average cases were correctly classified. Upon cross-validation, accuracy by group stayed consistent with the average group (82%) and improved with the gifted group (64%). CONCLUSIONS: Bioelectrical energy patterns of the nervous system, as measured by the AMI, could distinguish between gifted and average boys. The accuracy found in this current study using the AMI as a screening tool to detect giftedness is promising, however, further studies will need to be conducted to see if the findings can be replicated. PMID- 14736359 TI - Empathy, enablement, and outcome: an exploratory study on acupuncture patients' perceptions. AB - OBJECTIVES: To conduct an exploratory, retrospective study of acupuncture patients' perceptions of practitioner empathy, patient enablement, and health outcome, and to investigate the associations between them. METHODS: In a retrospective, observational study, questionnaires were distributed to 192 patients randomly selected from a population of 6348 who, several months previously, had participated in a survey of acupuncture safety, and had agreed to be contacted again. The main measures included patients' perceptions of their practitioners' empathy using the Consultation and Relational Empathy Measure, the Patient Enablement Instrument, and the Glasgow Homeopathic Hospital Outcome Scale (measuring change in main complaint and well-being). RESULTS: A total of 143 (74%) patients responded (27% men and 73% women) with an average age of 51 years. Comparisons between the population, the sample selected, and the responding sample showed reasonable equivalence. The majority of patients (71%) were in the middle of an ongoing course of treatment at the time of completing the questionnaires for this study. 36% of patients were attending for reasons of "general well-being," 34% for musculoskeletal problems, 11% for emotional or psychological problems, and 19% for other reasons. Empathy and enablement scores were not influenced by age or reason for attendance, but men showed significantly lower scores than women (p < 0.05). Patient enablement was significantly positively correlated with perception of their practitioners' empathy (Spearman's rho = 0.256, p < 0.01). Enablement in turn was strongly positively correlated with the outcome of both the main complaint (rho = 0.457, p < 0.0001) and improved well-being (rho = 0.521, p < 0.0001). CONCLUSION: Patients' perceptions of consultations with their acupuncturists suggest that their experience of empathy is significantly associated with patient enablement, which in turn is highly correlated with improved self-reported health outcomes. PMID- 14736360 TI - Review of sangre de drago (Croton lechleri)--a South American tree sap in the treatment of diarrhea, inflammation, insect bites, viral infections, and wounds: traditional uses to clinical research. AB - OBJECTIVE: The objective of this review is to provide an overview of the pharmacologic evidence that may or may not support clinical and ethnomedical uses of the sap of sangre de drago (dragon's blood; Croton lechleri Mull. Arg.). Data sources used were BIOSIS, EMBASE, PubMed, TOXLIT, International Pharmaceutical Abstracts, manual searches, papers on file from peer-reviewed journals, textbooks available at Armana Research, Inc., and researchers in the field of South American botanical medicine. CONCLUSIONS: The results of in vitro and in vivo studies largely support the majority of ethnomedical uses of sangre de drago including the treatment of diarrhea, wounds, tumors, stomach ulcers, herpes infection, the itching, pain and swelling of insect bites, and other conditions. Clinical studies of sangre de drago products have reported positive results in the treatment of traveler's and watery diarrhea and the symptoms of insect bites. Because the sap has shown low toxicity and preparations used in clinical studies were well tolerated, further clinical and pharmacologic studies are anticipated. Acknowledgment of the diversity in the chemical makeup of the sap from one geographic area to another and the recent characterization of alkaloid chemotypes of sangre de drago will require that materials developed for clinical use are standardized. PMID- 14736361 TI - Effect of 1-oxo-5beta, 6beta-epoxy-witha-2-ene-27-ethoxy-olide isolated from the roots of Withania somnifera on stress indices in Wistar rats. AB - OBJECTIVE: Isolation of biologically active fractions and compounds from the roots of Withania somnifera, a plant used extensively as a constituent of rasayana, in Ayurveda and to test their adaptogenic activity on stress indices using the cold-hypoxia-restraint (C-H-R) model. DESIGN: Bioactivity-guided fractionation of an aqueous extract of the roots of Withania somnifera led to the isolation of a new species of withanolide 1-oxo-5beta, 6beta-epoxy-witha-2-ene-27 ethoxy-olide. Structure elucidation, was carried out using proton nuclear magnetic resonance, infrared (IR), ultraviolet (UV), and mass spectroscopic analysis. Stress-related indices were evaluated, namely serum creatine phosphokinase (CPK) activity, serum lactate dehydrogenase (LDH) activity, serum corticosterone levels, and serum lipid peroxidation (LPO) levels. RESULTS: There was a significant decrease in a serum CPK, LDH, and LPO levels in animals pretreated with (1) fraction-I (20 mg/kg body weight), (2) 1-oxo-5beta, 6beta epoxy-witha-2-ene-27-ethoxy-olide (2.5 mg/kg body weight) in comparison to control when subjected to C-H-R stress. CONCLUSIONS: The results show that the a new species of withanolide, 1-oxo-5beta, 6beta-epoxy-witha-2-ene-27-ethoxy-olide (compound-1) could prove to be an effective agent to counteract C-H-R stress. PMID- 14736362 TI - Analysis of responses to valerian root extract in the feline pulmonary vascular bed. AB - OBJECTIVES: This study was undertaken to investigate pulmonary vascular response to valerian (Valeriana officinalis) in the feline pulmonary vasculature under constant flow conditions. DESIGN: In separate experiments, the effects of NG-L nitro-L-arginine methyl ester (L-NIO), a nitric oxide synthase inhibitor, glibenclamide, an adenosine triphosphate (ATP)-sensitive potassium (K+) channel blocker, meclofenamate, a nonselective cyclooxygenase (COX) inhibitor, bicuculline, a GABA(A) receptor antagonist, and saclofen, a GABA(B) antagonist, were investigated on pulmonary arterial responses to various agonists in the feline pulmonary vascular bed. These agonists included valerian, muscimol, a GABA(A) agonist, SKF-97541 a GABA(B) agonist, acetylcholine (ACh), and bradykinin, both inducers of nitric oxide synthase, arachidonic acid, a COX substrate, and pinacidil, an ATP-sensitive K+ channel activator, during increased tone conditions induced by the thromboxane A2 mimic, U46619. SETTINGS/LOCATION: Laboratory investigation. SUBJECTS: Mongrel cats of either gender. INTERVENTIONS: Injections of the abovementioned agonists and antagonists were given. OUTCOME MEASURES: Baseline pulmonary tone, responses to the agonists, and responses to the agonists after injections of antagonists were all measured via a pulmonary catheter transducer and recorded. RESULTS: Valerian root extract is a potent smooth muscle dilator in the feline pulmonary vascular bed. The vasodilatory effects of valerian root extract were unchanged after the administration of L NIO, glibenclamide, and meclofenamate. These effects were ablated, however, by both saclofen and bicuculline. The ability of saclofen and bicuculline to modulate the dilatory effects of valerian root extract was not statistically different. CONCLUSIONS: The vasodilatory effects of valerian root extract are mediated by a nonselective GABA mechanism. PMID- 14736363 TI - Extended Network Generalized Entanglement Theory: therapeutic mechanisms, empirical predictions, and investigations. AB - Extended Network Generalized Entanglement Theory (Entanglement Theory for short) combines two earlier theories based on complexity theory and quantum mechanics. The theory's assumptions are: the body is a complex, self-organizing system (the extended network) that self-organizes so as to achieve genetically defined patterns (where patterns include morphologic as well as lifestyle patterns). These pattern-specifying genes require feedback that is provided by generalized quantum entanglement. Additionally, generalized entanglement has evolved as a form of communication between people (and animals) and can be used in healing. Entanglement Theory suggests that several processes are involved in complementary and alternative medicine (CAM). Direct subtle therapy creates network change either through lifestyle management, some manual therapies, and psychologically mediated effects of therapy. Indirect subtle therapy is a process of entanglement with other people or physical entities (e.g., remedies, healing sites). Both types of subtle therapy create two kinds of information within the network- either that the network is more disregulated than it is and the network then compensates for this error, or as a guide for network change leading to healing. Most CAM therapies involve a combination of indirect and direct therapies, making empirical evaluation complex. Empirical predictions from this theory are contrasted with those from two other possible mechanisms of healing: (1) psychologic processes and (2) mechanisms involving electromagnetic influence between people (biofield/energy medicine). Topics for empirical study include a hyperfast communication system, the phenomenology of entanglement, predictors of outcome in naturally occurring clinical settings, and the importance of therapist and patient characteristics to outcome. PMID- 14736364 TI - Themes of holism, empowerment, access, and legitimacy define complementary, alternative, and integrative medicine in relation to conventional biomedicine. AB - Complementary and alternative medicine (CAM) has been defined largely in relation to conventional biomedicine. CAM therapies that are used instead of conventional medicine are termed "alternative." CAM therapies used alongside conventional medicine are said to be "complementary." "Integrative medicine" results from the thoughtful incorporation of concepts, values, and practices from alternative, complementary, and conventional medicines. The evolving process of integration between CAM and conventional medicine evokes new conceptual frameworks, as well as new terminology. Interview-based qualitative research at the University of Wisconsin-Madison seeks to probe and develop this theoretical structure. Interviews with users and practitioners of CAM therapies have revealed four primary themes: holism, empowerment, access, and legitimacy (HEAL). These themes characterize CAM and contrast it with conventional medicine. CAM is said to be more holistic and empowering yet less legitimate than conventional medicine. CAM is more intuitive; conventional is more deductive. While CAM is perhaps more psychologically accessible to many patients in that it better reflects commonly held values, it is often less financially and institutionally accessible, at least for those with conventional health insurance and limited income. Substantive barriers--including economic, organizational and scientific differences, as well as an apparent widespread lack of understanding--continue to thwart attempts at integration. More and better evidence is needed if CAM therapies are to be accepted by mainstream medicine. State-of-the-art research methods developed by conventional science will be needed to test CAM therapies. Conventional medicine, however, has much to learn from CAM. By incorporating a more holistic, empowering and accessible therapeutic approach, conventional medicine could build on its present legitimacy, and thereby enhance its power to "HEAL." PMID- 14736365 TI - Participant-centered analysis in complementary and alternative medicine comparative trials. AB - Participant-centered analysis involves applying the customary methods of statistical decision making at the level of the individual research participant. Consequently, each individual is declared a responder who benefited, a nonresponder, or possibly a responder who was harmed, using intensively collected data that were specific to that individual. There are several implications of the participant-centered approach. More data actually relevant to the important outcome need to be collected on individuals. The study results can be summarized in a simple table of responders/nonresponders by treatment group, and probabilities of true response can be estimated. The actual nature of the data collected and the statistical models used to analyze them drop into the background. Finally, production of individual-level decisions permits standard statistical approaches to be applied to the issue of which modality should be recommended for which person, instead of focusing on average effects and which modality should be recommended for everyone. PMID- 14736366 TI - Incorporating patients' perspectives in complementary and alternative medicine clinical trial design. AB - OBJECTIVES: To describe the importance and process of gathering the perspectives of former patients when designing clinical studies for complementary and alternative medicine (CAM) therapies and to describe how this information was used to guide the planning of a clinical study using complementary modalities to reduce symptom distress and enhance quality of life during the autologous stem cell transplantation (ASCT) process. DESIGN: Structured interviews with former ASCT patients to identify preferences, opinions, and other issues that may affect a clinical study in this population. SETTING: University of Virginia Health System Stem Cell Transplant Clinic. SUBJECTS: Ten (10) patients who had undergone ASCT within the previous year. RESULTS: In general, the 10 study participants interviewed reported that they would have been more receptive to receiving gentle Swedish massage than using guided imagery tapes during the ASCT process, although neither modality would have been particularly welcome during those treatment phases with highest physical or emotional/mental stress. Personal experiences, treatment side-effects, "personality," and life situation all had an influence on not only what was considered most stressful for the patient but also why it was perceived as stressful. CONCLUSION: Eliciting the views of persons who have undergone significant medical events is a necessary step in rigorous clinical trial development aimed at testing the efficacy of CAM modalities for symptom management. Consideration of patient preferences and motivations may ensure the best fit between interventions and the desired outcomes. PMID- 14736367 TI - From the five blind men to Cochrane Complementary Medicine systematic reviews. PMID- 14736369 TI - Selective genotyping for QTL detection using sib pair analysis in outbred populations with hierarchical structures. AB - A simulation study illustrates the effects of the inclusion of half-sib pairs as well as the effects of selective genotyping on the power of detection and the parameter estimates in a sib pair analysis of data from an outbred population. The power of QTL detection obtained from samples of sib pairs selected according to their within family variance or according to the mean within family variance within half sib family was compared and contrasted with the power obtained when only full sib pair analysis was used. There was an increase in power (4-16% ) and decrease in the bias of parameter estimates with the use of half-sib information. These improvements in power and parameter estimates depended on the number of the half sib pairs (half sib family size). Almost the same power as that obtained using all the available sib pairs could be achieved by selecting only 50-60% the animals. The most effective method was to select both full and half sib pairs on the basis of high within full sib family variance for the trait in question. The QTL position estimates were in general slightly biased towards the center of the chromosome and the QTL variance estimates were biased upwards, there being quite large differences in bias depending on the selection method. PMID- 14736370 TI - Frequencies of genes for coat colour and horns in Nordic cattle breeds. AB - Gene frequencies of coat colour and horn types were assessed in 22 Nordic cattle breeds in a project aimed at establishing genetic profiles of the breeds under study. The coat colour loci yielding information on genetic variation were: extension, agouti, spotting, brindle, dun dilution and colour sided. The polled locus was assessed for two alleles. A profound variation between breeds was observed in the frequencies of both colour and horn alleles, with the older breeds generally showing greater variation in observed colour, horn types and segregating alleles than the modern breeds. The correspondence between the present genetic distance matrix and previous molecular marker distance matrices was low (r = 0.08 - 0.12). The branching pattern of a neighbour-joining tree disagreed to some extent with the molecular data structure. The current data indicates that 70% of the total genetic variation could be explained by differences between the breeds, suggesting a much greater breed differentiation than typically found at protein and microsatellite loci. The marked differentiation of the cattle breeds and observed disagreements with the results from the previous molecular data in the topology of the phylogenetic trees are most likely a result of selection on phenotypic characters analysed in this study. PMID- 14736371 TI - Structure genetique des cecidomyies des cereales en Tunisie. AB - Genetic structure of Hessian flies in Tunisia. The genetic structure of M. destructor and M. hordei was investigated by sampling 21 fields of cereals in 14 localities of central and southern Tunisia. As previously shown, there was no strict association between the cereal species (wheat, barley and oat) and the Mayetiola species. M. destructor males displayed no heterozygosity at the Pgm3 locus, indicating that they were hemizygous as is the PGM locus in North America. In M. hordei, heterozygous males were observed at all loci, but strong heterozygote deficits were found at two loci (Mdh2 et Hk). Since no such deficit was observed in females, the population structure of M. hordei was studied only in females. Although heterozygosity was two fold higher in M. hordei than in M. destructor, the two species were similar for other genetic characteristics, including a low (Fst<0.05) but significant (P<0.05) genetic differentiation, no isolation by distance, and similar rates of gene flow (5.7 or=1ng x ml(-1)) was observed at 66 days (+/ 22 days s.d.) for the group of easy-calving multiparous suckling cows. Estimates of heritability and repeatability were h2 = 0.12 and r = 0.38 respectively, for the interval from calving to first oestrus (ICO). Corresponding values were h2 = 0.35 and r = 0.60 for the interval from calving to the first positive progesterone test (ICP). The genetic correlation between both criteria was high (rg = 0.98). The genetic relationships between postpartum intervals and BW and BCS of the female at calving were negative: the genetic aptitude to be heavier at calving and to have high body reserves was related to shorter postpartum intervals. A favourable genetic correlation between age at puberty and postpartum intervals was found (rg between 0.45 and 0.70). The heifers which were genetically younger at puberty also had shorter postpartum intervals. PMID- 14736387 TI - Estimation of genetic parameters for growth, carcass and overfeeding traits in a white geese strain. AB - In an experimental strain of white plumage geese created in 1989, two experiments were carried out from 1993 to 1995 in order to estimate genetic parameters for growth, and carcass composition traits in non-overfed animals, and genetic parameters for growth and fatty liver formation in overfed animals. Four hundred and thirty-one non-overfed animals were bred and slaughtered at 11 weeks of age; they were measured for forearm length, keel bone length, chest circumference and breast depth before and after slaughtering. The carcasses were partly dissected in order weigh breast, breast muscle and skin + fat, and abdominal fat. Four hundred and seventy-seven overfed animals were slaughtered at 20 weeks of age; they were measured for "paletot" (breast meat, bone and meat from wings, bone and meat from thigh and legs) weight and liver weight. In these two experiments, the weights had moderate to high heritability values. Breast depth measured on live animals showed a low heritability value. In overfed animals, liver weight showed a high heritability value. Liver weight could be increased by selection without a great effect on "paletot" weight. Thus, obtaining a white plumage geese strain for fatty liver production by selection would be difficult because only 20% of overfed animals had fatty liver. The results did not allow to conclude on the influence of selection on liver weight on carcass traits such as muscle or fatty tissue weight. PMID- 14736388 TI - Genetic parameters of body weight and prolificacy in pigeons. AB - Genetic parameters of body weight at weaning and of prolificacy were estimated in three commercial lines of pigeons selected by BLUP (Best Linear Unbiased Prediction) on both traits. The model of analysis took into account the direct genetic effects for both traits and the effect of parental permanent environment for body weight. Depending on the line considered, body weight varied from 556.7 g to 647.6 g and prolificacy ranged from 12.5 to 16.8 pigeons weaned per couple of parents per year. Heritability of body weight was high, varying between 0.46 and 0.60, and permanent environment was responsible for 6% to 9% of the total variability. On the contrary, prolificacy was poorly heritable (0.04 to 0.12). They were highly and negatively correlated (-0.77 to -0.82). Body weight showed significant genetic trends in lines B and C. No significant genetic difference could be observed between males and females for both traits. PMID- 14736389 TI - Cytogenetical anchoring of sheep linkage map and syntenic groups using a sheep BAC library. AB - In order to simultaneously integrate linkage and syntenic groups to the ovine chromosomal map, a sheep bacterial artificial chromosome (BAC) library was screened with previously assigned microsatellites using a sheep-hamster hybrid panel and genetic linkage. Thirty-three BACs were obtained, fluorescently labelled and hybridised on sheep-goat hybrid metaphases (2n = 57). This study allowed us, (i), to anchor all linkage groups on sheep chromosomes, (ii), to give information on the probable position of the centromere on the linkage map for the centromeric chromosomes, (iii), to contradict the previous orientation of the ovine X linkage group by the mapping of BMS1008 on OARXq38. Concerning our somatic cell hybrid panel, this study resulted in the assignment of all the previously unassigned groups to ovine chromosomes and a complete characterisation of the hybrid panel. In addition, since hybridisations were performed on a sheep goat hybrid, new marker/anchoring points were added to the caprine cytogenetic map. PMID- 14736390 TI - Mating schemes for optimum contribution selection with constrained rates of inbreeding. AB - The effect of non-random mating on genetic response was compared for populations with discrete generations. Mating followed a selection step where the average coancestry of selected animals was constrained, while genetic response was maximised. Minimum coancestry (MC), Minimum coancestry with a maximum of one offspring per mating pair (MC1) and Minimum variance of the relationships of offspring (MVRO) mating schemes resulted in a delay in inbreeding of about two generations compared with Random, Random factorial and Compensatory mating. In these breeding schemes where selection constrains the rate of inbreeding, DeltaF, the improved family structure due to non-random mating increased genetic response. For schemes with DeltaF constrained to 1.0% and 100 selection candidates, genetic response was 22% higher for the MC1 and MVRO schemes compared with Random mating schemes. For schemes with a less stringent constraint on DeltaF or more selection candidates, the superiority of the MC1 and MVRO schemes was smaller (5-6% ). In general, MC1 seemed to be the preferred mating method, since it almost always yielded the highest genetic response. MC1 mainly achieved these high genetic responses by avoiding extreme relationships among the offspring, i.e. fullsib offspring are avoided, and by making the contributions of ancestors to offspring more equal by mating least related animals. PMID- 14736391 TI - Effect of linkage on the control of inbreeding in selection programmes. AB - Selection and mating methods for controlling inbreeding in selection programmes are based on relationships obtained from pedigrees. The efficiency of these methods has always been tested by studies using genetic models of independent loci. However, under linkage the rate of inbreeding obtained from pedigrees can be different from the probability of identity by descent of genes. We simulated a quantitative trait under artificial selection controlled by a large number of genes spread on genome regions of different sizes. A method to control inbreeding based on minimising the average coancestry of selected individuals with a restriction in the loss of selection response, and a mating procedure to control inbreeding were applied. These methods, that use coancestry relationships, were not effective in controlling inbreeding when the genome sizes were smaller than five morgans or so. However, for larger genome sizes the methods were sufficiently efficient. For very tight linkage, methods that utilise molecular information from markers should be used. We finally discuss the effects of the selection of individual major genes on the neutral variability of adjacent genome regions. PMID- 14736392 TI - Predicting observed and latent responses to BLUP selection involving logistic variates. AB - Multivariate BLUPs can be derived when data are a mixture of continuous traits and observed discrete traits controlled by logistic latent traits. Algorithms were developed for predicting discrete responses to BLUP selection, and latent responses when the selection process included additional culling on scores. These algorithms were Taylor expansions using well-known expressions such as the probabilities and the two first moments of the truncated multinormal distribution, after appropriate re-parametrizations. They were compared to very accurate quadrature integrations. The test examples were suggested by a situation found in chickens where selection can involve body weight and leg deformity described by two logistic latent variates. Quadratic Taylor expansions generally provided a good accuracy. Therefore, they could be recommended when quadrature methods are too demanding, e.g., for complex breeding schemes. PMID- 14736393 TI - Genetic similarity and relationships of DNA fingerprints with performance and with heterosis in Japanese quail lines from two origins and under reciprocal recurrent or within-line selection for early egg production. AB - DNA fingerprints of Japanese quail male and female pure line breeders were obtained with probes 33.6, 33.15, and R18.1 and they yielded a total of 59 scoreable bands. Bandsharing (080-90% of the IgG was removed. This increase in detection of allergen-specific IgE occurred in sera from all mouse strains and to all allergens tested. In addition, reactivity of IgE antibodies to peanut or shrimp allergens by immunoblot increased visually approximately 4- to 10-fold. CONCLUSIONS: These studies indicate that allergen-specific IgG antibodies, which may be in more than 100-fold excess to IgE antibodies, interferes with detection of allergen-specific IgE, probably by competitive binding to allergenic epitopes. Substantial depletion of IgG antibodies (>80%) result in a significant increase in the sensitivity of the antibody measurements. PMID- 14736412 TI - Flow cytometry analysis of immune cell populations isolated from cervicovaginal secretions of cynomolgus monkeys. AB - Flow cytometric analysis was used in this study to characterize the lymphocyte population present in the vaginal mucosa of the cynomolgus monkey. Vaginal immune cells were obtained, using absorbent wicks, from 11 normal cycling female monkeys at different stages of the menstrual cycle and from three nursing monkeys (not cycling). Leucocytes, including lymphocytes and monocyte-macrophage cells, were present in the cervicovaginal secretions of healthy cynomolgous primates throughout the three phases of the menstrual cycle. We also found that even if immune cells were constant throughout the menstrual cycle, among the T cell subsets there were differences. CD8+ cells [14.5+/-9% (mean+/-S.D.); range 3-30%] were more numerous compared to the mean number of CD4+ cells [7.3+/-5% (mean+/ S.D.); range 2-15%]. Characterization of the vaginal cells during the nursing period showed that the monocyte-macrophage (CD14+, CD11c+) cells were abundant compared with the low number of both B (CD20+) and T cells (CD2+). Our results show that cytometric analysis by FACS can be used to identify the immune cell populations present at the local level. This technique may provide a useful tool by which the vaginal environment can be studied in order to correlate cell phenotype with immune function. PMID- 14736413 TI - Characterization and performance evaluation of in vivo and in vitro produced monoclonal anti-TNT antibodies for the detection of TNT. AB - In this paper are the experimental results used to characterize four distinct monoclonal anti-TNT antibodies (in vivo and in vitro cloned) for potential use in a field-portable immunosensor. Direct and competitive enzyme-linked immunosorbent assays (ELISA) were performed to determine their affinity for TNT and a fluorescently labeled analog of TNT for use in an immunosensor. Effective concentrations (EC(50)), inhibition concentration (IC(50)) and cross-reactivity measurements to related nitroaromatics (e.g., 2,4,6-trinitrobenzene [TNB], methyl 2,4,6-trinitrophenyl nitramine [tetryl], 2-amino-4,6-dinitrotoluene [2A-4,6-DNT], 2,4-dinitrotoluene [2,4-DNT] and 1,3-dinitrotoluene [1,3-DNT]) were measured. Final characterization of the monoclonal antibodies was based on performance (measured by fluorescence dose response) using a fluorescence-based microcapillary displacement assay. Analytical techniques showed a high degree of affinity for TNT and varying degrees of cross-reactivity with each respective monoclonal antibody. Microcapillary displacement immunoassays with each of the antibodies resulted in detection capabilities at the lowest applied TNT concentration (10 ng/ml). PMID- 14736415 TI - Sodium chloride fixation of tissues under field conditions in tropical countries. AB - Fixation of skin and lymph node fragments in anhydric sodium chloride at room temperature for periods of weeks or months was found to preserve morphological structures and immunoreactivity. Fragments of a size of 6-8x6x6 mm were dried and placed in anhydric sodium chloride powder in sealed containers. Prior to further processing, the specimens were desalinated and rehydrated, then snap-frozen, sectioned, fixed in acetone and stained with monoclonal antibodies. The intensity of staining of molecular structures did not differ from that of primarily snap frozen specimens. The method allows fixation of tissues under field conditions in tropical countries and at places lacking freezing facilities. PMID- 14736414 TI - Microsphere-based duplexed immunoassay for influenza virus typing by flow cytometry. AB - We have developed a rapid, duplexed microsphere-based immunoassay for the characterization of influenza virus types that has the potential to overcome many of the limitations of current detection methods. The assay uses microspheres of two sizes, each coupled to an influenza type A- or type B-specific monoclonal antibody (MAb), to capture influenza viruses in the sample. A cocktail of fluorescently labeled, influenza-specific polyclonal antibodies then binds the captured viruses. The sandwich complexes are measured using a multiparameter flow cytometer. The assay can distinguish between influenza types A and B in a single reaction with good reproducibility and high sensitivity. Detection sensitivity is much higher than that of commercially available influenza diagnosis quick kits: the FLU OIA (Thermo Biostar) kit and the Directigen Flu A+B kit (Becton Dickinson). The multiplexing capabilities of the current assay, which are not possible with enzyme-linked immunosorbent assay (ELISA) and the commercially available kits, reduce sample handling and consume fewer costly reagents. This assay represents a more efficient and sensitive method of characterizing influenza types. With inclusion of influenza subtype-specific antibodies as capture antibodies, this microsphere-based immunoassay can be expanded to differentiate among influenza types and subtypes in a single reaction to improve world-wide influenza surveillance. PMID- 14736416 TI - Efficient purification of unique antibodies using peptide affinity-matrix columns. AB - Phage display technology was used to identify peptide ligands with unique specificity for a monoclonal model antibody, MK16, that recognises the human multiple sclerosis associated MHC class II molecule DR2 in complex with a myelin basic protein (MBP)-derived peptide corresponding to residue 85-99. Several peptide epitopes were identified and all of them recognised specifically MK16. One peptide, ER6.1, was selected and linked to beaded agarose and demonstrated excellent performance as a peptide affinity chromatography matrix. This epitope matrix was efficient in the purification of MK16 Fab fragments and had no affinity for other antibodies. Using this peptide matrix MK16 IgG could be purified from cell culture supernatants thereby separating MK16 IgG from bovine IgG normally present in the enriched growth media used for such cells. Investigations of the fine specificity of the ER6.1 peptide demonstrated that it recognised a unique epitope within the heavy chain CDR3 region of the MK16 antibody. Thus, variants of MK16 antibody, which had retained the specificity and affinity of the original antibody but had slightly different amino acid composition in the CDR3 region, were not recognised by the ER6.1 peptide. PMID- 14736417 TI - Simultaneous isolation of platelet factor 4 and glycoprotein IIb-IIIa complex from rabbit platelets, and characterization of specific chicken antibodies to assay them. AB - Rabbits are frequently used as models for studying coagulation and platelet disorders. However, few reports on literature have dealt with the purification and characterization of rabbit platelet proteins. Herein a protocol for the simultaneous purification of rabbit platelet factor 4 (PF4) and platelet glycoprotein IIb-IIIa (GPIIb-IIIa, integrin alpha(IIb)beta(3)) is described. Specific antibodies were raised in laying chicken, which were used for assaying PF4 by ELISA, and GPIIb-IIIa by direct immunofluorescence and flow cytometry. Furthermore, the binding of monoclonal antibodies specific for GPIIb-IIIa complex (P2), ligand-induced binding site of GPIIIa (LIBS1) and rabbit P-selectin (12A7), as well as of polyclonal IgY specific for rabbit GPIIb-IIIa, was compared in quiescent and thrombin-activated platelets. Polyclonal anti-rabbit PF4 IgY was a specific and sensitive probe that could be used for assaying PF4 in plasma samples. GPIIb-IIIa expression was increased in thrombin-activated platelets, as evaluated by flow cytometric analysis using P2 and polyclonal antibodies raised in chickens. Rabbit GPIIb-IIIa also exhibited a conformational modification that caused the appearance of ligand-induced binding sites. Increased P-selectin expression, used as a positive control, was also noticeable in thrombin-activated platelets. These data evidence that antibodies raised in laying chickens specific to rabbit PF4 and GPIIb-IIIa, as well as certain monoclonal antibodies specific for human GPIIb-IIIa, may be used for investigating rabbit platelet physiology. PMID- 14736418 TI - Monitoring dendritic cells in clinical practice using a new whole blood single platform TruCOUNT assay. AB - Dendritic cells (DC) from distinct DC subsets are essential contributors to normal human immune responses. Despite this, reliable assays that enable DC to be counted precisely have been slow to evolve. We have now developed a new single platform flow cytometric assay based on TruCOUNT beads and the whole blood "Lyse/No-Wash" protocol that allows precise counting of the CD14(-) blood DC subsets: CD11c(+)CD16(-) DC, CD11c(+)CD16(+) DC, CD123(hi) DC, CD1c(+) DC and BDCA-3(+) DC. This assay requires 50 microl of whole blood; does not rely on a hematology blood analyser for the absolute DC counts; allows DC counting in EDTA samples 24 h after collection; and is suitable for cord blood and peripheral blood. The data is highly reproducible with intra-assay and inter-assay coefficients of variation less than 3% and 11%, respectively. This assay does not produce the DC-T lymphocyte conjugates that result in DC counting abnormalities in conventional gradient-density separation procedures. Using the TruCOUNT assay, we established that absolute blood DC counts reduce with age in healthy individuals. In preliminary studies, we found a significantly lower absolute blood CD11c(+)CD16(+) DC count in stage III/IV versus stage I/II breast carcinoma patients and a lower absolute blood CD123(hi) DC count in multiple myeloma patients, compared to age-matched controls. These data indicate that scientific progress in DC counting technology will lead to the global standardization of DC counting and allow clinically meaningful data to be obtained. PMID- 14736419 TI - Quantitation of dengue virus specific CD4+ T cells by intracellular cytokine staining. AB - We developed an intracellular cytokine staining assay to quantify dengue specific memory T cells elicited by a primary dengue virus (DEN) infection. Peripheral blood mononuclear cells (PBMC) of volunteers who received experimental live attenuated monovalent DEN vaccines were stimulated with glutaraldehyde inactivated dengue-infected Vero cell culture lysates from all four DEN serotypes. CD4+ T cell frequencies to previously identified MHC class II peptides were equivalent to 40-70% of the responses using virus infected cell lysates in two donors. IFN-gamma responses to DEN were detected from 0.04% to 0.45% CD4+ T cells. The highest IFN-gamma response was elicited by antigens from the homologous serotype. We detected serotype-specific and -cross reactive CD4+ T cell cytokine responses from all donors. This assay is suitable for measuring DEN specific CD4+ T cells in human PBMC from large population studies where donor haplotype is unknown or highly variant. PMID- 14736420 TI - Direct comparison of traditional ELISAs and membrane protein arrays for detection and quantification of human cytokines. AB - Many labs wish to measure cytokines in an accurate, reproducible, and rapid manner. An antibody-based membrane array for measuring cytokines has been developed based on the same technology as the traditional ELISA. The aim of this study was to compare results obtained with the traditional ELISA method with those from the membrane array technology, a form of low-cost proteomics. Diluted human whole blood was stimulated with live bacteria (Escherichia coli, or Staphylococous aureus), or LPS and cytokines were measured both by ELISA and the membrane protein array. Of the 16 cytokines measured via ELISA, only IFN-gamma was below detection level. The other 15 cytokines were present in concentrations up to several thousand picograms/ml. Of the 20 cytokines measured via membrane protein array, only 3 could be detected (IL-6, IL-8 and MIP-1beta). Additionally, the membrane protein array did not detect TNF-alpha from the LPS-stimulated blood. These results indicate that the low-cost membrane protein array may lack sufficient sensitivity to adequately detect cytokines levels in complex biological fluids such as human plasma. PMID- 14736421 TI - Immuno-SLM-a combined sample handling and analytical technique. AB - Immuno-supported liquid membrane (immuno-SLM) extraction is a new technique that makes use of antibody (Ab)-antigen interactions as the "extraction force" to drive the mass transfer in a selective way. In immuno-SLM, anti-analyte (Ag) Abs are introduced into the acceptor phase of the SLM unit to trap the Ag that passes from the flowing donor through the SLM into the stagnant acceptor. The amount of immuno-extracted analyte (AbAg) is quantified by connecting the immuno-SLM unit on-line with a non-competitive heterogeneous fluorescence flow immunoassay (FFIA) that makes use of a fluorescein-labeled analyte tracer that titrates the residual excess of Ab present in the acceptor. A restricted access (RA) column is used for the separation of the two tracer fractions (Ag* and AbAg*) formed, and the eluted AbAg* fraction is measured downstream by a fluorescence detector. Factors influencing the optimum immuno-SLM extraction parameters, i.e., donor flow rate, extraction time and type of Ab, were investigated for immuno extraction of the model analyte atrazine. Immuno-SLM coupled to FFIA (immuno-SLM-FFIA) and FFIA alone were compared in terms of the assay sensitivities obtained and the sample matrix influence. The concentration at the mid-point of the calibration curve (IC(50)) was 16.0+/-1.4 and 36+/-16 microg/l, the limit of detection (LOD) was 2.0+/-1.1 and 20+/-10 microg/l, and the dynamic range was 2-100 and 20-500 microg/l atrazine for immuno-SLM-FFIA and FFIA, respectively. The matrix influence on the FFIA was significant in orange juice and surface water, whereas the influence was minor for immuno-SLM-FFIA with recoveries between 104% and 115% for 5 microg/l atrazine in tap water, orange juice and river water. PMID- 14736422 TI - Bivalent antibody phage display mimics natural immunoglobulin. AB - We report the development of a system for displaying bivalent antibody fragments on M13 bacteriophage in a manner that effectively mimics the binding behavior of natural antibodies. In the "bivalent display" format, two copies of antigen binding sites are displayed on the coat of a single phage particle. Bivalent display was first achieved by the insertion of a dimerization domain, consisting of an IgG1 hinge region and a homodimerizing GCN4 leucine zipper, between a Fab and the C-terminal domain of the M13 gene-3 minor coat protein. In a phagemid based display system, the resulting "Fab'-zip-phage" particles display bivalent Fabs that resemble natural IgGs. An important functional consequence of bivalent display is an avidity effect, which results in a greatly reduced off-rate for phage bound to immobilized antigen. The avidity effect improved the capture and retention of bivalent Fab'-zip-phage relative to monovalent Fab-phage both with antigen immobilized on plates and with cell surface antigen. To examine the requirements for bivalent display on phage, we systematically trimmed down the dimerization domain and found that a single cysteine was sufficient to confer the same avidity effect conferred by the complete dimerization domain. Bivalent antibody phage display should be useful for many applications. In particular, the technology should aid in the production of antibodies against difficult antigens, and also, in selections that require dimerization for activity. PMID- 14736423 TI - Efficient gene transfer into the human natural killer cell line, NKL, using the Amaxa nucleofection system. AB - Natural killer (NK) cell lines are useful for studying facets of NK cell biology. Such cell lines are notoriously difficult to transfect by traditional methods, a fact that has hampered NK cell biology studies for a long time. To overcome this, we investigated the use of the Amaxa nucleofection system that directly transfers DNA into the nucleus of the cell. This technology has revolutionized transfection studies with heretofore relatively transfection resistant cell types such as T cells, B cells and dendritic cells. Despite these advances, NK cells and NK cell lines have remained relatively resistant to transfection, including nucleofection. In this study we employed cDNA for SHP1 and various Rab proteins cloned in enhanced green/yellow fluorescent protein (EGFP/EYFP) expression plasmids for transient transfections into NKL cells. The expression of EGFP/EYFP fusion proteins was analyzed by flow cytometry, immunoblot and confocal microscopic analyses. We achieved 40-70% transfection efficiency with high levels of expression in this cell line with 85-90% viability. The method used in this report proves to be far superior to existing methods for delivering DNA into this well studied NK cell line and, consequently, provides new experimental opportunities. PMID- 14736424 TI - An ultrasensitive method for the detection of oligoclonal IgG bands. AB - We have developed an ultrasensitive isoelectrofocusing (IEF) method for the detection of oligoclonal (OGC) IgG bands in cerebrospinal fluid (CSF) and sera. In this procedure, double antibody and peroxidase immunodetection have been substituted by a single antibody labelled with alkaline phosphatase. Alkaline phosphatase immunodetection not only improves tenfold the sensitivity of the assay but also gives a sharper pattern resolution making the interpretation easier and increases the specificity of this technique. This preliminary report should be validated in a larger number of patients with and without multiple sclerosis (MS). PMID- 14736425 TI - Selection of scFvs specific for HBV DNA polymerase using ribosome display. AB - We applied a ribosome display technique to a mouse scFv library to select single chain variable fragments (scFvs) specific for the terminal protein (TP) of hepatitis B virus (HBV) DNA polymerase. Synthetic TP-peptide was used as an antigen to obtain scFvs that recognize specific epitopes within the TP domain, the priming site of HBV DNA polymerase. The scFv DNA library was transcribed in vitro to mRNA for ribosome display. scFv-ribosome-mRNA complexes were produced using a rabbit reticulocyte lysate system, and were panned against TP-peptide under appropriate conditions. After each panning, putative scFv-encoding genes were recovered by RT-PCR, and the analysis showed that scFv-ribosome-mRNA complexes were specifically selected by the TP-peptide. We used a radioimmunoassay to show that the TP-peptide-specific scFv pools were enriched through the selection process. Selected scFvs showed binding activity for both the TP-peptide and the HBV DNA polymerase protein in an ELISA. Sequence analysis showed that each TP-specific scFv had a different sequence, and that random mutagenesis was mediated by ribosome display. PMID- 14736426 TI - Affinity-independent elution of antibody-displaying phages using cleavable DNA linker containing streptavidin beads. AB - The search for high-affinity antibodies from phage libraries by panning is often complicated by inefficient detachment of the captured phage particles from the antigen-coated solid phase. General elution methods, independent on the bioaffinity interaction, would be desirable for high-throughput antibody screening. We have developed a bioaffinity-independent elution method based on the use of commercially available streptavidin paramagnetic particles. PMID- 14736427 TI - Production of a biotinylated single-chain antibody fragment in the cytoplasm of Escherichia coli. AB - Biotinylated antibodies are commonly used reagents in research and molecular diagnostics. The traditional approach to biotinylate antibodies is to conjugate a chemically active biotin derivative to certain chemical groups on protein surface. An alternative method, which can be used for site-specific biotinylation of recombinant antibodies, takes advantage of the capability of the enzyme biotin ligase to catalyze the attachment of a biotin to a unique lysine residue in specific protein/peptide substrates that can be genetically linked to the antibody to generate a fusion protein. We describe here expression of functional scFv and concomitant enzymatic biotinylation of it in bacterial cytoplasm. The anti-thyroid-stimulating hormone (TSH) scFv was produced as an N-terminal fusion with the biotinylated domain of the biotin carboxyl carrier protein of Escherichia coli in the redox modified E. coli strain Origami B which has an oxidizing cytoplasmic environment. After optimization of the biotin concentration and expression temperature, this approach allowed the production of biotinylated and immunoreactive fusion protein with the yield of 1.4 mg/l/OD(600) (13.6 mg/l) in a simple shake flask culture. The biotinylated fusion protein released from disrupted cells can be directly used, for example, in immunoassay applications. This was proved by setting up a TSH immunoassay using the bio-scFv as a solid phase capture antibody. The sensitivity of the assay was comparable with the currently used commercial immunoassays. PMID- 14736428 TI - Detection of differential expression of mouse interferon-alpha subtypes by polymerase chain reaction using specific primers. AB - Specific primers for nine mouse interferon-alpha (IFN-alpha) subtypes, namely, IFN-alpha1, IFN-alpha1-9, IFN-alpha2, IFN-alpha4, IFN-alpha5, IFN-alpha7, IFN alpha6/8, IFN-alpha11, and IFN-alphaB, were designed and evaluated on Poly(I).Poly(C)-induced and influenza virus-infected L929 cells. Specificity of the primers was confirmed in a cross-polymerase chain reaction (cross-PCR). IFN alpha1, IFN-alpha1-9, IFN-alpha4, IFN-alpha6/8, IFN-alpha11, and IFN-alphaB were found to be induced in L929 cells 6-9 h after Poly(I).Poly(C) treatment. The amplification of a particular subtype was not biased in the presence of excess of other templates. Differential expression of the IFN-alpha subtypes was observed in influenza A/NWS/33- and B/Lee/40-infected L929 cells. A/NWS/33 virus was found to upregulate the gene expression of IFN-alpha1, IFN-alpha4, IFN-alpha6/8, IFN alpha11, and IFN-alphaB in L929 cells as early as 6 h after infection. In B/Lee/40-infected L929 cells, only IFN-alpha4 was upregulated. Our results suggest that the designed primers will serve as a useful tool in analyzing the expression of IFN-alpha subtypes in various systems and hence for the evaluation of their function. PMID- 14736429 TI - AMEGA: antigen-mediated genetically modified cell amplification. AB - Selection of genetically modified cells is a critical step to engineer the cells with desired properties. While antibiotic selection has been commonly used, administration of cytotoxic drugs often leads to deleterious effects not only to inert cells but also to transfected or transduced ones. To overcome this problem, a positive screening method for genetically modified cells is proposed using a pair of chimeric receptors that trigger a growth signal in response to a specific antigen. Either V(H) or V(L) region of anti-hen egg lysozyme (HEL) antibody HyHEL 10 was fused to extracellular D2 domain of erythropoietin receptor (EpoR) and transmembrane/cytoplasmic domains of either EpoR or gp130. A model transgene, enhanced green fluorescent protein (EGFP) and the chimeric receptor genes that reconstituted functional Fv were retrovirally co-infected to interleukin (IL)-3 dependent Ba/F3 cells, followed by direct HEL selection in the absence of IL-3. Consequently, a single round of selection led to a single population of EGFP positive cells. The detailed protocol of the method termed antigen-mediated genetically modified cell amplification (AMEGA) is described. PMID- 14736430 TI - What is the optimal serum potassium level in cardiovascular patients? AB - Humans are prone to sodium overload and potassium depletion. This electrolyte imbalance is important in the pathogenesis of cardiovascular disease and sudden cardiac death. Avoiding hypokalemia is beneficial in several cardiovascular disease states including acute myocardial infarction, heart failure, and hypertension. The evidence highlighting the importance of potassium homeostasis in cardiovascular disease and possible mechanisms explaining potassium's benefits are reviewed. Targets for serum potassium concentration are suggested. PMID- 14736431 TI - Eptifibatide provides additional platelet inhibition in non-ST-elevation myocardial infarction patients already treated with aspirin and clopidogrel. Results of the platelet activity extinction in non-Q-wave myocardial infarction with aspirin, clopidogrel, and eptifibatide (PEACE) study. AB - OBJECTIVES: The present study hypothesis was that eptifibatide offered further antiplatelet efficacy above clopidogrel in non-ST-elevation myocardial infarction (NSTEMI) patients before an expeditive coronary intervention. BACKGROUND: Although thienopyridines and glycoprotein (GP) IIb/IIIa antagonists are often co prescribed in the context of NSTEMI, the antiplatelet interaction of these agents is poorly described and the superiority of GP IIb/IIIa antagonists above thienopyridine treatment alone is not clear. METHOD: Thirty-two NSTEMI patients treated with aspirin and enoxaparin were studied using flow cytometry to define parameters of platelet activation with a panel of agonists before clopidogrel, after clopidogrel, and during an eptifibatide infusion following the clopidogrel load. RESULTS: After platelet activation with adenosine diphosphate, thrombin receptor-activating peptide, or U46-619, relative reductions in conformationally activated GP IIb/IIIa receptor expression (evaluated with PAC-1) of 48%, 43%, and 33%, respectively (all p < 0.0001), were seen with clopidogrel, but further 80%, 78%, and 72% (all p < 0.0001) reductions were seen with eptifibatide. With the same agonists, fibrinogen binding was significantly reduced after clopidogrel by 70%, 64%, and 81% (all p < 0.0001) and again further reduced with eptifibatide by 90%, 95%, and 69% (all p < 0.0001). The total number of GP IIb/IIIa receptors (measured as P2 expression) and P-selectin expression fell after clopidogrel, after ex vivo stimulation with the same agonists; however, both parameters increased slightly during the eptifibatide infusion. CONCLUSIONS: The activated GP IIb/IIIa expression and fibrinogen binding findings indicate that eptifibatide provides significant potent antiplatelet activity above aspirin and clopidogrel, suggesting additive immediate protection in the treatment of NSTEMI. The P2 and P selectin findings suggest the possibility of a partial agonist and/or pro inflammatory effect. PMID- 14736432 TI - Adverse outcome in aortic sclerosis is associated with coronary artery disease and inflammation. AB - OBJECTIVES: The present study was designed to evaluate the relationship between the presence of aortic sclerosis, serologic markers of inflammation, and adverse cardiovascular outcomes. BACKGROUND: Aortic sclerosis is associated with adverse cardiovascular outcomes. However, the mechanism by which such nonobstructive valve lesions impart excess cardiovascular risk has not been delineated. METHOD: In 425 patients (mean age 68 +/- 15 years, 54% men) presenting to the emergency room with chest pain, we studied the relationship among aortic sclerosis, the presence and acuity of coronary artery disease, serologic markers of inflammation, and cardiovascular outcomes. Patients underwent echocardiography and serologic testing including C-reactive protein (CRP). Aortic valves were graded for the degree of sclerosis, and cardiovascular outcomes including cardiac death and nonfatal myocardial infarction (MI) were analyzed over one year. RESULTS: Aortic sclerosis was identified in 203 patients (49%), whereas 212 (51%) had normal aortic valves. On univariate analysis at one year, patients with aortic sclerosis had a higher incidence of cardiovascular events (16.8% vs. 7.1%, p = 0.002) and worse event-free survival (normal valves = 93%, mild aortic sclerosis = 85%, and moderate to severe aortic sclerosis = 77%, p = 0.002). However, by multivariable analysis aortic sclerosis was not independently associated with adverse cardiovascular outcomes; the only independent predictors of cardiac death or MI at one year were coronary artery disease (hazard ratio [HR] 3.23, p = 0.003), MI at index admission (HR 2.77, p = 0.008), ascending tertiles of CRP (HR 2.2, p = 0.001), congestive heart failure (HR 2.15, p = 0.02) and age (HR 1.03, p = 0.04). CONCLUSIONS: The increased incidence of adverse cardiovascular events in patients with aortic sclerosis is associated with coronary artery disease and inflammation, not a result of the effects of valvular heart disease per se. PMID- 14736433 TI - Why is aortic sclerosis associated with adverse clinical outcomes? PMID- 14736434 TI - Impaired brachial artery endothelium-dependent and -independent vasodilation in men with erectile dysfunction and no other clinical cardiovascular disease. AB - OBJECTIVES: The goal of this study was to determine whether patients with vascular erectile dysfunction (ED) and no other clinical cardiovascular disease have structural and functional abnormalities of other vascular beds. BACKGROUND: In many ED patients, vascular disease is the major underlying cause. It may be that ED is an early marker of atherosclerosis in patients without clinical cardiovascular disease. METHOD: We assessed systemic vascular structure and function in 30 patients with ED and 27 age-matched normal control (NL) subjects. We measured vascular parameters, including: 1) carotid and brachial artery diameters, intima-media thickness, compliance, and distensibility; 2) aortic pulse wave velocity; 3) coronary calcification; and 4) brachial artery endothelium-dependent and -independent vasodilation. RESULTS: There were no significant differences in baseline demographics, coronary artery risk score, or lipid values between the two groups. Most structural and functional vascular parameters were similar in the ED and NL groups. Brachial artery flow-mediated vasodilation (FMD) (1.3 vs. 2.4%, p = 0.014) and vasodilation to nitroglycerin (NTG) (13.0 vs. 17.8%, p < 0.05) were significantly reduced in ED patients, compared with NL subjects. In addition, there was a significant correlation between FMD and vasodilation to NTG in ED patients (r = 0.59, p < 0.05) but not in NL subjects. CONCLUSIONS: Patients with ED but no clinical cardiovascular disease have a peripheral vascular defect in endothelium-dependent and independent vasodilation that occurs before the development of other overt functional or structural systemic vascular disease and is independent of other traditional cardiovascular risk factors. PMID- 14736435 TI - Erectile dysfunction: the earliest sign of generalized vascular disease? PMID- 14736436 TI - Prevalence of specific variant carotid geometric patterns and incidence of cardiovascular events in older persons. The Cardiovascular Health Study (CHS E 131). AB - OBJECTIVES: We hypothesized that variant geometric patterns of the common carotid artery (CCA) predict the incidence of cardiovascular disease (CVD), after accounting for CCA intima-medial thickness (IMT). BACKGROUND: Common carotid artery intima-media thickness has been associated with the incidence of cardiovascular disease. METHOD: Noninvasive measurements of IMT were made with high-resolution ultrasonography in 5,640 subjects 65 years of age or older participating in the Cardiovascular Health Study. New coronary and/or cerebrovascular events served as outcome variables over a median 10.2-year follow up. To characterize different carotid structural geometric patterns (CGP), vascular mass (VM) was combined with the wall-to-lumen ratio (W/L). Normal values for W/L and VM were defined as age-adjusted, gender-specific 75th percentiles of the 1,899 normotensive subjects free of CVD at baseline. Four CGPs were defined: CGP1 = normal W/L ratio and VM; CGP2 = arterial remodeling (i.e., increased W/L ratio with normal VM); CGP3 = arterial hypertrophy (i.e., increased W/L ratio with increased VM); and CGP4 = arterial hypertrophy with dilation (i.e., normal W/L ratio and increased VM). RESULTS: Coronary or cerebrovascular events (adjusted for age, gender, traditional risk factors, and IMT) were associated with CGP in subjects free of CVD at baseline. Specifically, the hazard ratio (Cox proportional-hazards analyses) for CGP3 (arterial hypertrophy) was 1.25 (95% confidence interval [CI] 1.03 to 1.53), and for CGP4 (arterial hypertrophy with dilation) was 1.43 (95% CI 1.16 to 1.75) compared with CGP1 (normal). CONCLUSIONS: Arterial hypertrophy defined by variant CGP patterns is associated with the development of new CVD, independent of age, traditional risk factors, and CCA IMT. PMID- 14736437 TI - Utility of myocardial perfusion imaging in patients with low-risk treadmill scores. AB - OBJECTIVES: The purpose of this study was to determine whether a previously validated clinical score (CS) could identify patients with a low-risk Duke treadmill score who had a higher risk of adverse events and, therefore, in whom myocardial perfusion imaging would be valuable for risk stratification. BACKGROUND: Current American College of Cardiology/American Heart Association guidelines recommend using a standard exercise test without imaging as the initial test in patients who have an interpretable electrocardiogram and are able to exercise. METHOD: We studied 1,461 symptomatic patients with low-risk Duke treadmill scores (> or =5) who underwent myocardial perfusion imaging. The CS was derived by assigning one point to each of the following variables: typical angina, history of myocardial infarction, diabetes, insulin use, male gender, and each decade of age over 40 years. A CS cutoff > or =5 or <5 was used to categorize patients as high risk (n = 303 [21%]) or low risk (n = 1,158 [79%]). Perfusion scans were categorized as low, intermediate, or high risk on the basis of the global stress score (GSS). RESULTS: High-risk scans were more common in patients with a high-risk CS (26.4% vs. 9.5%, p < 0.0001). The CS and GSS were significant independent predictors of cardiac death. However, in patients with a low CS, seven-year cardiac survival was excellent, regardless of the GSS (99% for normal scans, 99% for mildly abnormal scans, and 99% for severely abnormal scans). In contrast, patients with a high CS had a lower seven-year survival rate (92%), which varied with GSS (94% for normal scans, 94% for mildly abnormal scans, and 84% for severely abnormal scans; p < 0.001). CONCLUSIONS: In symptomatic patients with low-risk Duke treadmill scores and low clinical risk, myocardial perfusion imaging is of limited prognostic value. In patients with low risk Duke treadmill scores and high clinical risk, annual cardiac mortality (>1%) is not low, and myocardial perfusion imaging has independent prognostic value. PMID- 14736438 TI - Stress myocardial perfusion single-photon emission computed tomography is clinically effective and cost effective in risk stratification of patients with a high likelihood of coronary artery disease (CAD) but no known CAD. AB - OBJECTIVES: We sought to evaluate the prognostic and cost implications of stress myocardial perfusion single-photon emission computed tomography (SPECT), or MPS, in patients with a high pretest likelihood (>0.85) of coronary artery disease (CAD) with no previous CAD. BACKGROUND: Sparse data are available regarding the prognostic performance characteristics of MPS in this patient group. METHOD: We followed up 1,270 consecutive patients with no previous revascularization or myocardial infarction (MI), with a pre-exercise tolerance test (ETT) likelihood of CAD > or =0.85, who underwent exercise or adenosine stress MPS (follow-up 94.4% complete; 2.2 +/- 1.2 years; 60 hard events [5.9%, 2.6%/year]). Risk adjustment of survival data was done using Cox proportional hazards analysis. Costs per reclassification of risk were calculated using assumed costs and threshold analyses. RESULTS: In patients treated medically after MPS, normal MPS had a low risk of cardiac death and hard events (0.6% and 1.3% per year, respectively). With increasing extent and severity of MPS defects, the risk of both cardiac death and hard events increased significantly (p < 0.05). Cox models indicated that the addition of MPS data resulted in incremental prognostic value over pre-MPS data (chi-square increase 48 to 87, p < 0.0001). Compared with strategies of initial referral to ETT in patients able to exercise, initial referral to MPS appeared to be a more cost-effective strategy. Similarly, compared with a strategy of direct referral to catheterization in patients with a high likelihood of CAD, initial referral to MPS is a cost-saving approach. CONCLUSIONS: In patients with a high likelihood of CAD but without known CAD, stress MPS yields incremental value and achieves risk stratification in a cost effective manner. The current results support a strategy of initial stress imaging in this patient cohort, as a reasonable alternative to direct referral to catheterization or initial ETT. PMID- 14736439 TI - Risk stratification using stress myocardial perfusion imaging: don't neglect the value of clinical variables. PMID- 14736440 TI - Technetium 99m sestamibi myocardial perfusion imaging predicts clinical outcome in the community outpatient setting. The Nuclear Utility in the Community (NUC) Study. AB - OBJECTIVES: The purpose of this study was to evaluate the prognostic value of community-based myocardial perfusion imaging (MPI) and to assess the incremental value of individual components of (99m)Tc-sestamibi single photon emission computed tomography (SPECT). BACKGROUND: Although the most rapid growth of MPI has been in community outpatient laboratories, its prognostic value has not been validated in this setting. METHOD: We prospectively followed 1,612 consecutive patients undergoing stress (99m)Tc-sestamibi SPECT in an outpatient community laboratory who experienced 71 hard events over 24 +/- 7 months (0.2% lost to follow-up). RESULTS: Patients whose scans were normal incurred an annualized event rate of 0.4%, compared with 2.3% for those with abnormal scans (p < 0.0001). Subset analysis demonstrated comparable risk stratification for women and men, diabetics, patients with normal resting ECGs, and those referred for pharmacologic and exercise stress. After adjusting for pre-test variables, multivariable Cox regression analysis found the most potent independent components of MPI to be, in order of importance, transient ischemic dilation, extent of reversibility, post-stress ejection fraction, extent and severity of the stress perfusion defect, and the overall test result (normal or abnormal). Each 1% decrement of ejection fraction predicted a 3% increase in risk (p = 0.0009). Post-MPI angiography and revascularization increased commensurate with the extent and severity of MPI result. CONCLUSIONS: The prognostic value of perfusion imaging is portable and transferable to the outpatient community setting, with multiple components of MPI providing incremental prognostic information. PMID- 14736441 TI - Myocardial positron emission computed tomographic images obtained with fluorine 18 fluoro-2-deoxyglucose predict the response of idiopathic dilated cardiomyopathy patients to beta-blockers. AB - OBJECTIVES: The aim of this study was to elucidate whether the response of idiopathic dilated cardiomyopathy (DCM) patients to beta-blockers can be predicted by positron emission tomography with fluorine-18 fluoro-2-deoxyglucose (FDG-PET). BACKGROUND: Patients with DCM often have a poor prognosis, and it is important to predict their response to beta-blocker therapy, which may be effective in DCM. However, no accurate methods of predicting their response have been available. METHOD: In 22 DCM patients with reduced left ventricular (LV) systolic function, FDG-PET was performed. Uptake in the LV after glucose loading was evaluated based on the average global percent uptake of the injected dose (G%ID) and the coefficient of variance (CV) in 24 segments of the LV. Uptake during fasting was evaluated semiquantitatively on the basis of the total uptake score. The beta-blocker was administered, and LV function was monitored by echocardiography. The histologic findings were assessed in the 18 patients who underwent endomyocardial biopsy. RESULTS: The beta-blocker was effective in the majority of patients whose G%ID after glucose loading was >0.7%, and the sensitivity and specificity of G%ID as a predictor of beta-blocker efficacy were 83.3% and 90.0%, respectively. Percent CV did not predict efficacy. Four groups, defined on the basis of the FDG uptake score during fasting and G%ID after glucose loading, had distinctive histologic findings and outcomes. CONCLUSIONS: It has been shown that FDG-PET is a good predictor for the effectiveness of beta blockers. PMID- 14736442 TI - Acute comparative effect of right and left ventricular pacing in patients with permanent atrial fibrillation. AB - We tested the hypothesis that left ventricular (LV) pacing is superior to right ventricular (RV) apical pacing in patients undergoing atrioventricular (AV) junction ablation and pacing for permanent atrial fibrillation. The potential benefit of LV over RV pacing needs to be evaluated without the confounding effect of other variables that can influence cardiac performance. An acute intrapatient comparison of the QRS width and echocardiographic parameters between RV versus LV pacing was performed within 24 h after ablation in 44 patients. Both modes of pacing were also compared with pre-implantation values. Compared with RV pacing, LV pacing caused a 5.7% increase in the ejection fraction (EF) and a 16.7% decrease in the mitral regurgitation (MR) score; the QRS width was 4.8% shorter with LV pacing. Similar results were observed in patients with or without systolic dysfunction and/or native left bundle branch block, except for a greater improvement in MR in the latter group. Compared with pre-ablation measures, the EF increased by 11.2% and 17.6% with RV and LV pacing, respectively; the MR score decreased by 0% and 16.7%; and the diastolic filling time increased by 12.7% and 15.6%.Rhythm regularization achieved with AV junction ablation improved EF with both RV and LV pacing; LV pacing provided an additional modest but favorable hemodynamic effect, as reflected by a further increase of EF and reduction of MR. The effect seems to be equal in patients with both depressed and preserved systolic functions and in those with and without native left bundle branch block. PMID- 14736443 TI - Pacing the left ventricle: does underlying rhythm matter? PMID- 14736444 TI - Effect of rate or rhythm control on quality of life in persistent atrial fibrillation. Results from the Rate Control Versus Electrical Cardioversion (RACE) Study. AB - OBJECTIVES: We studied the influence of rate control or rhythm control in patients with persistent atrial fibrillation (AF) on quality of life (QoL). BACKGROUND: Atrial fibrillation may cause symptoms like fatigue and dyspnea. This can impair QoL. Treatment of AF with either rate or rhythm control may influence QoL. METHOD: Quality of life was assessed in patients included in the Rate Control Versus Electrical Cardioversion for Persistent Atrial Fibrillation (RACE) study (rate vs. rhythm control in persistent AF). Rate control patients (n = 175) were given negative chronotropic drugs and oral anticoagulation. Rhythm control patients (n = 177) received serial electrocardioversion, antiarrhythmic drugs, and oral anticoagulation, as needed. Quality of life was studied using the Short Form (SF)-36 health survey questionnaire at baseline, one year, and the end of the study (after 2 to 3 years of follow-up). At baseline, QoL was compared with that of healthy control subjects. Patient characteristics related to QoL changes were determined. RESULTS: Mean follow-up was 2.3 years. At baseline, QoL was lower in patients than in age-matched healthy controls. At study end, under rate control, three subscales of the SF-36 improved. Under rhythm control, no significant changes occurred compared with baseline. At study end, QoL was comparable between both groups. The presence of complaints of AF at baseline, a short duration of AF, and the presence of sinus rhythm (SR) at the end of follow up, rather than the assigned strategy, were associated with QoL improvement. CONCLUSIONS: Quality of life is impaired in patients with AF compared with healthy controls. Treatment strategy does not affect QoL. Patients with complaints related to AF, however, may benefit from rhythm control if SR can be maintained. PMID- 14736445 TI - Intra-left ventricular electromechanical asynchrony. A new independent predictor of severe cardiac events in heart failure patients. AB - OBJECTIVES: We sought to assess the electromechanical parameters, using tissue Doppler echocardiography, as potential independent predictors of heart failure (HF) worsening. BACKGROUND: Ventricular conduction disorders worsen the prognosis for HF patients. However, the relationships between the QRS width and morphology, hemodynamic parameters, and presence and magnitude of intra-left ventricular (LV) and inter-ventricular (V) asynchrony have not been well clarified. METHOD: A total of 104 patients with an LV ejection fraction (EF) 4 cm) liver metastases, patients undergoing cryotherapy combined with liver resection and patients receiving blood transfusion intraoperatively, especially when exceeding 4 units packed red cells, had a significantly impaired survival in univariate analysis. In multivariate analysis (Cox regression) the amount of intraoperative blood transfusion was the only independent prognostic indicator. CONCLUSION: Intraoperative blood transfusion has a negative impact on survival following hepatic cryotherapy for colorectal liver metastases and should be avoided by refinement of surgical technique whenever possible. Patients with multiple liver metastases or metastases of more than 4 cm in size have an impaired prognosis-therefore trials of adjuvant therapies following hepatic cryotherapy for colorectal liver metastases should include number and size of metastases for stratification of patient groups. PMID- 14736519 TI - Elective surgery for colorectal cancer in a defined Swedish population. AB - AIMS: The aim of this study was to describe variability in compliance to clinical guidelines in colorectal cancer surgery related to hospital structure. METHODS: All patients registered in the databases of the Regional Oncologic Centre, operated upon electively for colon cancer between the start of the register in 1997 until 2000 (n=1771) and for rectal cancer between the start of the register in 1995 until 2000 (n=1841) were selected for analysis. RESULTS: There was no difference in 5-year survival rate between colon and rectal cancer (mean follow up 2.6 and 3.0 years, respectively; p=0.22). There was a significant difference in frequency of preoperative liver scan depending on hospital category with an increase in colon cancer from 39 to 46% (p=0.02) and in rectal cancer from 42 to 64% (p<0.001). For colon cancer there was no difference, according to hospital category, in quotient sigmoid and high anterior resection to left-sided resection. Furthermore, high anterior resection was more common at university and general district hospitals (8%) compared with district hospitals (4%) (p=0.01). Sphincter-saving surgery was more common at university hospitals and district general hospitals than at district hospitals (low anterior/abdomino-perineal resection quotients 2.3, 2.4 and 1.6, respectively; p<0.001). CONCLUSIONS: Population-based audit forms an appropriate and valuable basis for quality assurance projects. In addition to describing compliance to guidelines and pointing to process steps that can be improved, such investigations may also indicate changes due to scientific development. Linked to case-costing data, such results may form an important basis for decisions about modifications in health care. PMID- 14736521 TI - Predicting survival in patients with liver cancer considered for transarterial chemoembolization. AB - INTRODUCTION: Transarterial chemoembolization (TACE) has been used extensively to treat tumours confined to the liver in patients unsuitable for surgical resection. This study attempts to identify patients with liver cancer most likely to benefit from this type of treatment. PATIENTS AND METHODS: All patients undergoing TACE for liver cancer between 1989 and 2001 were included in the study. RESULTS: In a group of 137 consecutive patients undergoing TACE, univariate analysis identified a number of pre-treatment factors that were associated with poor prognosis. Multivariate analysis of these factors subsequently identified three pre-treatment factors; age greater than 60, serum alkaline phosphatase concentration >120U/l and albumin less than 35 g/l; that were independently and significantly associated with reduced survival duration. A scoring system was devised with one point allocated for each adverse factor which produced median survivals related to points scored as follows, 0 points-20 months, 1 point-12 months, 2 points-7 months and 3 points-4 months. To validate this scoring system the next 40 consecutive patients undergoing TACE were studied prospectively. These patients had median survival durations related to points scored as follows 0 points not calculable, 1 point-10 months, 2 points-7 months, 3 points-4 months. CONCLUSION: This simple scoring system can be used to predict prognosis in patients with liver cancer and may assist in clinical decision making in the selection of patients likely to benefit from TACE. PMID- 14736522 TI - The role of transjugular intrahepatic portosystemic shunt prior to abdominal tumoral surgery in cirrhotic patients with portal hypertension. AB - INTRODUCTION: Major abdominal surgery can be contraindicated in some cirrhotic patients because of severe portal hypertension. The present study reports our experience of three patients with abdominal tumours prepared for surgery by transjugular intrahepatic portosystemic shunts (TIPS) in order to reduce portal hypertension and the risk of intraoperative bleeding. PATIENTS AND METHODS: Three patients with cirrhosis and portal hypertension diagnosed with a right colon carcinoma, an adenocarcinoma of pancreas and a gastric and sigmoid synchronic tumours in the same patient. Because portal hypertension was the leading cause of surgical contraindication, neoadjuvant TIPS placement was proposed before surgery. RESULTS: TIPS placement was performed without intra-procedure complications. An average reduction of 18 mmHg was achieved in portosystemic gradients. The planned operations were performed with a delay of 14-45 days after TIPS without intraoperative bleeding. Complications occurred in one patient without operative mortality. CONCLUSION: TIPS placement allows a pre-operative portal decompression in cirrhotic patients with portal hypertension and abdominal tumours that require surgical treatment. This procedure reduces the risk of bleeding by reducing the portosystemic gradient and the varices around the tumoral area. This procedure is less invasive than conventional shunt surgery, but it is not free of complications and should be performed by experienced interventional radiologists on selected patients. This is still an experimental indication of TIPS which efficacy must be confirmed in larger series. PMID- 14736523 TI - CD8+ lymphocyte infiltration and apoptosis in hepatocellular carcinoma. AB - AIMS: Tumour-infiltrating lymphocytes (TILs) play a role in local anti-tumour immunity. Tumour cells may escape from immune surveillance by expressing RCAS1, a receptor-binding cancer antigen expressed on SiSo cells, which inhibits T cell growth. In this study, the correlation between the density of CD8+ TILs, tumour cell apoptosis, and tumour RCAS1 expression was investigated in hepatocellular carcinoma (HCC). METHODS: We obtained tissues from 60 patients with surgically resected HCCs. CD8+ TILS, apoptotic cancer cells, and RCAS1 expressing cancer cells were identified by immunohistochemistry. RESULTS: The density of CD8+ T cells in tumours (mean: 9.5/HPF, HPF: high power field) was significantly less than in non-cancerous hepatic lobules (17.8/HPF, p<0.001) and in relation to the progression of tumour stage. The density of CD8+ T cells in tumours positively correlated with the occurrence of tumour cell apoptosis, but did not correlate with RCAS1 protein expression. CONCLUSIONS: CD8+ TILs may play a role in the occurrence of tumour cell apoptosis in HCC, but CD8+ TILs may not be controlled by RCAS1 in HCC. PMID- 14736524 TI - Risk factors for malignancy in multinodular goitres. AB - BACKGROUND: Multinodular goitre (MNG) is quite often associated with thyroid carcinoma, but the risk factors for malignancy are not well known. The aim is to analyse patients with thyroid carcinoma associated with MNG to determine the clinical risk factors for malignancy. METHOD: From a series of 672 MNGs we analysed a subgroup of 59 patients presenting with an associated thyroid carcinoma. The variables analysed were age, sex, family history, cervical radiation therapy, residence in areas of endemic goitre, prior thyroid surgery, time of evolution, asymptomatic status, hyperthyroidism, compressive syndromes, intrathoracic prolongation, goitre consistency and cervical adenopathies. These were compared to MNGs not developing malignancy using the Chi-squared test, Student's t test and a logistic regression test. RESULTS: Of the 59 cases, 37 corresponded to a microcarcinoma (< or =1 cm). The most common histological type was papillary (n=48), followed by follicular (n=6). In 20 cases the carcinoma was multifocal, and capsular involvement was noted in 16 patients, lymph node involvement in five and vascular involvement in another five. The multivariate analysis confirmed the following as independent variables associated with the presence of carcinoma: family history of thyroid pathology (RR=1.6), history of cervical radiation therapy (RR=1.8), recurrent goitre (RR=2.1) and presence of adenopathies on physical exploration (RR=1.6). CONCLUSION: The risk factors for carcinoma associated with MNG are family history of thyroid pathology, personal history of cervical radiation therapy, prior surgery and presence of cervical adenopathies. PMID- 14736525 TI - Surgery for lung metastases in Ewing's sarcoma of bone. AB - The addition of chemotherapy and radiotherapy to local treatment for Ewing's sarcoma of bone (ES) significantly bettered its prognosis. Nonetheless, although with no signs of local recurrence some patients still relapse with lung metastases. These patients are generally treated with non-surgical procedures. The few reports in literature on surgical resection of metachronous pulmonary metastases do not allow any conclusions. This study aims to report the results achieved in 24 Ewing's sarcoma patients who relapsed only with lung metastases, and no local recurrence, treated with surgery at the authors' institution. PMID- 14736526 TI - Rac1 gene mutations in human brain tumours. AB - AIMS: Rac1 is a member of the Ras superfamily of small GTPase and plays a fundamental role in cytoskeleton reorganization, regulation of gene expression and cell proliferation, and cellular transformation. Though recent studies point to an involvement of rac1 in tumorigenesis, little is known about the alteration of rac1 gene in human brain tumours. METHODS: Reverse transcription-polymerase chain reaction (RT-PCR), TA cloning, and DNA sequencing were performed to detect rac1 gene mutations in the surgical specimens of 45 human brain tumours. RESULTS: Twelve of 45 cases had base changes in the rac1 gene. The frequency of rac1 alterations was seven of 18 meningiomas, three of 14 astrocytomas, one of seven pituitary adenomas, and one of four metastatic brain tumours. No mutation was detected in acoustic neurilemomas. The subtypes of seven meningiomas include three meningotheliomatous, two atypical, one transitional and one angioblastic meningioma. Three astrocytomas had rac1 gene mutation, including one grade II, one grade III, and one grade IV astrocytoma. All of single base changes were transitions, five of them being T to C transitions. Sites of rac1 mutation were found in codons 34, 41 (two cases), 42 (two cases), 43, 44, 46 and 58. These mutations are mainly localized in the putative effector-domain of rac1 gene and may enhance the activity of rac1, which increases the survival of brain tumours. CONCLUSION: Our results suggest that rac1 gene may play a role in some brain tumours of divergent histogenesis and that the alterations of rac1 gene may contribute to tumorigenesis and/or metastasis. PMID- 14736527 TI - Topoisomerase II alpha and telomerase expression in papillary thyroid carcinomas. AB - BACKGROUND: Altered topoisomerase II alpha (Topo II alpha) expression and telomerase activity (TA) reflect tumour cell growth and malignant transformation. METHODS: We examined TA by using a TRAP assay and expression of Topo II alpha by immunohistochemical analysis in a series of 27 cases of papillary thyroid carcinoma (PTC). RESULTS: Topo II alpha labelling index (LI) ranged from 0.1 to 4.2% and was significantly associated with patient age (r=-0.42, p=0.003), with higher levels of Topo II alpha in patients under 40 years. There was no relationship between Topo II alpha LI, AGES score or other clinical outcome. TA was detected in 14 PTC, with relative levels ranging from 1.2 to 102 units. A significant positive correlation between the multiplicity of tumoral foci and the TA levels (p<10(-2)) was noted. CONCLUSION: We concluded that Topo II alpha cannot be used as a marker of tumour aggressiveness. Furthermore, enhanced Topo II alpha expression in PTCs from patients less than 40 years old suggests that this age group might benefit from Topo II inhibitor chemotherapy. PMID- 14736528 TI - Parapharyngeal metastases from thyroid cancer. AB - AIM: To emphasise the pattern of lymphatic dissemination in the parapharyngeal space from thyroid cancer. PATIENTS AND METHOD: Among 696 patients treated for thyroid cancer between 1986 and 2001, parapharyngeal metastasis was diagnosed in three patients, previously treated for papillary thyroid carcinoma. RESULTS: All three patients have been treated by surgical resection through lateral cervical approach. Two of them were controlled regionally whereas the remaining one had a submucosal pharyngeal metastasis locally resected 27 months after parapharyngeal resection. CONCLUSIONS: Parapharyngeal metastasis is rare, but should be a recognized pattern of lymphatic dissemination from thyroid carcinoma to avoid unnecessary radioiodine and because surgical resection is efficacious with acceptable morbidity. PMID- 14736529 TI - Necrotizing pancreatitis after radiofrequency destruction of pancreatic tumours. AB - BACKGROUND: Intraoperative radiofrequency (RF) can be used to treat multiple small pancreatic tumours. PATIENTS AND METHODS: Two patients with multiple pancreatic metastases from renal cancer were treated with RF destruction of the pancreatic tumours. The first was treated with a monopolar device and the second with a bipolar device. A high temperature (>90 degrees C) was used, and one patient underwent cooling of the intrapancreatic common bile duct. RESULTS: The destruction of tumours was effective. However, the two patients presented post operatively a severe necrotizing pancreatitis, with life-threatening hemorrhagic complications. CONCLUSION: RF destruction of pancreatic tumours is dangerous with current devices. PMID- 14736530 TI - Re: Epidermoid carcinoma arising in chronic lymphedema, Bilen et al. PMID- 14736534 TI - Impact factors: no totum pro parte by skewness of citation. PMID- 14736535 TI - LDL, IGF-1, and VSMC apoptosis: linking atherogenesis to plaque rupture in vulnerable lesions. PMID- 14736536 TI - Transgenic models in cardiac arrhythmias: how close can we get to the bedside? PMID- 14736537 TI - Electrophysiological changes in heart failure and their relationship to arrhythmogenesis. AB - This review focuses mainly on studies in non-ischemic animal models of heart failure. These animals develop ventricular arrhythmias, mostly non-sustained ventricular tachycardia, and often die suddenly. Clinical studies suggest that sudden death is due to ventricular tachycardia or fibrillation in about 50% of cases, the other half to bradyarrhythmias or electromechanical dissociation. Electrophysiologic changes in heart failure are not confined to the ventricles: the intrinsic sinus rate is reduced due to a downregulation of If and sensitivity to acetylcholine is enhanced by upregulation of the muscarinic receptor. Reduction of heart rate may be a protective mechanism since at rapid rates contractility is reduced and the likelihood for triggered activity due to delayed afterdepolarizations is enhanced. The beneficial effect of beta-adrenergic blockade in patients may be partly due to the reduction in sinus rate. Although the results of different studies often vary, the most consistent electrophysiological changes in the ventricles are prolongation of the action potential, especially at slow rates, a reduction in the transient outward current Ito, the rapid and slow components of the delayed rectifier Ikr and Iks, and the inward rectifier Ik1. Abnormalities in intracellular calcium handling play a major role in the genesis of delayed afterdepolarizations. Triggered activity based on delayed afterdepolarizations has been demonstrated in failing myocardium and are caused by spontaneous release of calcium from the sarcoplasmic reticulum (SR), especially in the presence of noradrenaline. Three factors combine to the enhanced propensity for the occurrence of delayed afterdepolarizations: (1) increased activity of the Na/Ca exchanger, (2) a reduced inward rectifier, (3) residual beta-adrenergic responsiveness required to raise the reduced sarcoplasmic calcium content to a level where spontaneous calcium release occurs. Early afterdepolarizations have also been demonstrated, especially in human myocytes from failing hearts in the presence of noradrenaline. Mapping experiments have shown that the ventricular arrhythmias are mainly due to non reentrant mechanisms, most likely triggered activity based on delayed afterdepolarizations. PMID- 14736538 TI - Metabolic remodelling of the failing heart: the cardiac burn-out syndrome? AB - It has been postulated that the failing heart suffers from chronic energy starvation, and that derangements in cardiac energy conversion are accessory to the progressive nature of this disease. The molecular mechanisms driving this 'metabolic remodelling' process and their significance for the development of cardiac failure are still open to discussion. Next to changes in mitochondrial function, the hypertrophied heart is characterized by a marked shift in substrate preference away from fatty acids towards glucose. It has been argued that the decline in fatty acid oxidation is not fully compensated for by a rise in glucose oxidation, thereby imposing an additional burden on overall ATP generating capacity. Several lines of evidence suggest that these metabolic adaptations are brought about, at least in part, by alterations in the rate of transcription of genes encoding for proteins involved in substrate transport and metabolism. Here, the principal metabolic changes are reviewed and the various molecular mechanisms that are likely to play a role are discussed. In addition, the potential significance of these changes for the aetiology of heart failure is evaluated. PMID- 14736539 TI - The endothelin system in pulmonary arterial hypertension. AB - Endothelin-1 (ET-1), a peptide produced primarily by vascular endothelial cells, was discovered in 1980 and it was characterized as a powerful vasoconstrictor and mitogen for smooth muscle. ET-1 binds to two types of receptors, ETA and ETB: ETA receptors are found in smooth muscle cells, whereas ETB-receptors are localized on both endothelial cells and in smooth muscle cells. Activation of ETA- and ETB receptors on smooth muscle cells mediates the vasoconstrictive and mitogenic effects of ET-1. Stimulation of endothelial ETB-receptors promotes ET-1 clearance and activation of NO and prostacyclin release. Pulmonary arterial hypertension (PAH) is a severe condition characterized by a progressive increase in pulmonary vascular resistance leading to right ventricular failure and death. An activation of the ET-1 system has been demonstrated in both plasma and lung tissues of PAH patients as well as in animal models of PAH. The most efficient way to antagonize the ET-1 system is the use of ET-1 receptor antagonists that can block either ETA or ETA- and ETB-receptors. These drugs are effective in animal models of PAH and have been tested in multiple clinical trials in patients with PAH. Bosentan, an orally active, dual ET-1 receptor antagonist has been shown to improve symptoms, exercise capacity, hemodynamics, echocardiographic parameters and the outcome of patients with severe PAH, and it has been approved for clinical use in many countries. The selective ETA-receptor antagonist sitaxentan has improved exercise capacity and hemodynamics of PAH patients in two preliminary studies. The main side effect of ET-1 antagonists is the increase of liver enzymes likely due to an accumulation of bile salts cytotoxic to hepatocytes. Additional trials with these drugs are currently ongoing. In conclusion, the hypothesis that the ET-1 system over-activation can be successfully antagonised in patients with PAH has been clearly demonstrated. PMID- 14736540 TI - Interactions between the sympathetic nervous system and the kidneys in arterial hypertension. AB - Elevated sympathetic activity changes renal function and accelerates the development of hypertension. Principles of sympatho-renal interactions in chronic hypertension are reviewed. Alterations in the ontogeny of the sympathetic nervous system and the kidney, inherited abnormalities in sensory receptor function and exaggerated responsiveness to mental stress contribute to inappropriately high sympathetic activity in primary or essential hypertension. Careful characterization of clinical study populations shows that elevated sympathetic activity and "essential" hypertension are not unequivocally associated. Prospective clinical studies which investigate a broader array of physiological functions and experiments in recombinant inbred rodents with less traumatic nerve recording techniques than currently available will help to define under which conditions elevated sympathetic activity is indeed a cause of primary hypertension. Signals arising from the kidney which activate the renin angiotensin system and afferent renal nerves increase sympathetic activity. These mechanisms importantly contribute to the pathogenesis of hypertension secondary to renal artery stenosis and end-stage renal disease. PMID- 14736541 TI - Native and oxidized low density lipoproteins oppositely modulate the effects of insulin-like growth factor I on VSMC. AB - OBJECTIVE: Changes in the local expression and signaling activity of the insulin like growth factor-I (IGF-I) axis regulate growth and survival of plaque-derived vascular smooth muscle cells (VSMC) and influence plaque fate. Recent evidence suggests that accumulation of low density lipoproteins (LDL) in VSMC during the progression of atherogenesis is linked to local changes in IGF-I signaling. We investigated the effects of LDL on the biological actions and downstream signaling pathways mediated by this growth factor in A10 VSMC. METHODS AND RESULTS: We first characterized the effects of LDL on the proliferative and anti apoptotic actions of IGF-I in A10 VSMC. Native LDL were mitogenic and synergistically enhanced DNA synthesis induced by IGF-I from 4-, 9- up to 7.8 fold, while having no effect on its anti-apoptotic actions. In contrast, oxidized LDL, at oxidation levels that did not modify these actions by themselves, significantly reduced the mitogenic and survival effects of IGF-I by 40% and 60%, respectively. These observations correlated with opposite changes exerted by native and oxidized LDL on the insulin receptor substrate-1 (IRS)-associated PI3 kinase/Akt response to IGF-I. The extracellular signal-regulated kinase (ERK) signaling response was not affected. CONCLUSIONS: Our study demonstrates a previously unidentified modulation of the actions of IGF-I on A10 VSMC by LDL, dependent on their extent of oxidative modification. Our findings suggest that the differential modulation of the PI3 kinase/Akt response to IGF-I play a pivotal role. PMID- 14736542 TI - Mechanisms by which SCN5A mutation N1325S causes cardiac arrhythmias and sudden death in vivo. AB - OBJECTIVE: Mutations in the cardiac sodium channel gene SCN5A are responsible for type-3 long QT disease (LQT3). The genesis of cardiac arrhythmias in LQT3 is multifaceted, and the aim of this study was to further explore mechanisms by which SCN5A mutations lead to arrhythmogenesis in vivo. METHODS: We engineered selective cardiac expression of a long QT syndrome (LQTS) mutation (N1325S) in human SCN5A and generated a transgenic mouse model, TGM(NS31). RESULTS: Conscious and unrestrained TGM(NS31)L12 mice demonstrated a significant prolongation of the QT-interval and a high incidence of spontaneous polymorphic ventricular tachycardia (VT) and fibrillation (VF), often resulting in sudden cardiac death (n=52:156). Arrhythmias were suppressed by mexiletine, a sodium channel blocker for the late persistent sodium current. Action potentials (APs) from TGM(NS31)L12 ventricular myocytes exhibited early afterdepolarizations and longer 90% AP durations (APD90=69 +/- 5.9 ms) than control (APD90=46.7 +/- 4.8 ms). Voltage clamp experiments in transgenic myocytes revealed a peak inward sodium current (INa) followed by a slow recovery from inactivation. After mexiletine application, transgenic ventricular APDs were shortened, and recovery from inactivation of INa was enhanced. These suggest that the N1325S transgene is responsible for the abnormal signals present in transgenic cells as well as the genesis of lethal arrhythmias in mice. Interestingly, transgenic but not wild type myocytes displayed longer APDs with a shortening of CLs. CONCLUSIONS: Our findings show that a new model for LQTS has been established, and we report on an arrhythmogenic mechanism that, unlike other SCN5A mutations, results in poor restitution of APD with increasing rate as a possible substrate for arrhythmogenesis. PMID- 14736543 TI - Prolonged repolarization and triggered activity induced by adenoviral expression of HERG N629D in cardiomyocytes derived from stem cells. AB - OBJECTIVE: The long QT syndrome, N629D HERG mutation, alters the pore selectivity signature sequence, GFGN to GFGD. Heterologous co-expression of N629D and the wildtype HERG resulted in a relative loss of the selectivity of K+ over Na+, but its physiologic relevance has not been assessed in cardiac myocytes. METHODS AND RESULTS: Accordingly, N629D was overexpressed, via adenoviral gene transfer, in cardiomyocytes derived from mouse stem cells. Three IKr phenotypes were observed: (1) the wildtype-like IKr showed inward rectification and a positive tail current; (2) the N629D-like IKr showed outward rectification and an inward tail current; and (3) intermediate IKr showed a small outward tail current. Action potentials (AP) were paired with the IKr measurements in each cell. Resting membrane potential (RMP) was critically dependent on the IKr phenotype. The resting membrane potential of the cells was -61 +/- 5 mV (n=40) in wildtype, -63 +/- 3 mV (n=18) in wildtype-like IKr phenotype, -30 +/- 2 mV (n=12) in N629D-like and -47 +/- 2 mV (n=24) in intermediate phenotype (p<0.00001). Triggered action potential durations (APD) were: 62 +/- 12 ms (n=6) in wildtype, 65 +/- 11 ms (n=6) in wildtype-like IKr phenotypes and 106 +/- 10 ms (n=6) (p<0.01) in intermediate IKr phenotypes. Lowering [K+]o hyperpolarized wildtype cells and cells with a wildtype-like IKr phenotype, but depolarized those with intermediate phenotype (from -45 +/- 1 to -35 +/- 0.5 mV (n=12), p<0.01). In 6 of 12 cells, with intermediate phenotype, the hypokalemia-induced depolarization resulted in triggered activity. TTX suppressed this triggered activity. CONCLUSION: Overexpression of N629D in cardiomyocytes derived from stem cells results in phenotypic variability in IKr, which was the critical determinant of the resting membrane potential, action potential duration and arrhythmogenic response to low [K+]o. PMID- 14736544 TI - Na+/Ca2+ exchange current (I(Na/Ca)) and sarcoplasmic reticulum Ca2+ release in catecholamine-induced cardiac hypertrophy. AB - OBJECTIVE: Catecholamines that accompany acute physiological stress are also involved in mediating the development of hypertrophy and failure. However, the cellular mechanisms involved in catecholamine-induced cardiac hypertrophy, particularly Ca2+ handling, are largely unknown. We therefore investigated the effects of cardiac hypertrophy, produced by isoprenaline, on I(Na/Ca) and sarcoplasmic reticulum (SR) function in isolated myocytes. METHODS: I(Na/Ca) was studied in myocytes from Wistar rats, using descending (+80 to -110 mV) voltage ramps under steady state conditions. Myocytes were also loaded with fura-2 and either field stimulated or voltage clamped to assess [Ca2+]i and SR Ca2+ content. RESULTS: Ca2+-dependent, steady state I(Na/Ca) density was increased in hypertrophied myocytes (P<0.05). Ca2+ release from the SR was also increased, whereas resting [Ca2+]i and the rate of decline of [Ca2+]i to control levels were unchanged. SR Ca2+ content, estimated by using 10.0 mmol/l caffeine, was also significantly increased in hypertrophied myocytes, but only when myocytes were held and stimulated from their normal resting potential (-80 mV) but not from -40 mV. However, the rate of decline of caffeine-induced Ca2+ transients or I(Na/Ca) was not significantly different between control and hypertrophied myocytes. Ca2+ dependence of I(Na/Ca), examined by comparing the slope of the descending phase of the hysteresis plots of I(Na/Ca) vs. [Ca2+]i, was also similar in the two groups of cells. CONCLUSION: Data show that SR Ca2+ release and SR Ca2+ content were increased in hypertrophied myocytes, despite an increase in the steady state I(Na/Ca) density. The observation that increased SR function occurred only when myocytes were stimulated from -80 mV suggests that Na+ influx may play a role in altering Ca2+ homeostasis in hypertrophied cardiac muscle, possibly through increased reverse Na+/Ca2+ exchange, particularly at low stimulation frequencies. PMID- 14736545 TI - Initial steps of insulin signaling and glucose transport are defective in the type 2 diabetic rat heart. AB - OBJECTIVE: Whole body insulin resistance and diabetes are risk factors for cardiovascular diseases, yet little is known about insulin resistance in the diabetic heart. The aim of this work was to define the insulin response in hearts of the Goto-Kakizaki (GK) rat, a polygenic model of spontaneous type 2 diabetes. METHODS: We measured D[2-3H]glucose uptake before and after insulin stimulation, plus initial steps of the insulin signaling pathway after insulin infusion via the caudal vena cava in hearts from the male Wistar and spontaneously diabetic GK rats. RESULTS: Despite normal basal D[2-3H]glucose uptake, insulin-stimulated glucose uptake was 50% (p<0.03) lower in GK rat hearts compared with their Wistar controls. Total GLUT4 protein was depleted by 28% (p<0.01) in GK rat hearts. We found 31% (p<0.0001) and 38% (p<0.001) decreased protein levels of insulin receptor beta (IRbeta)-subunit and insulin receptor substrate-1 (IRS-1), respectively, in GK rat hearts with 37% (p<0.02) and 45% (p<0.01) lower insulin stimulated tyrosine phosphorylation of these proteins. Owing to the decreased IRS 1 protein levels, GK rat hearts had a 41% (p<0.0001) decrease in insulin stimulated IRS-1 protein association with the p85 subunit of phosphatidylinositol 3-kinase, despite normal phosphatidylinositol 3-kinase protein expression. Insulin-stimulated serine phosphorylation of protein kinase B was the same in all hearts, as was protein kinase B expression. CONCLUSION: We conclude that decreased insulin receptor beta, IRS-1 and GLUT4 proteins are associated with insulin resistance in type 2 diabetic rat hearts. PMID- 14736546 TI - The development of myocardial insulin resistance in conscious dogs with advanced dilated cardiomyopathy. AB - BACKGROUND: The failing heart demonstrates a preference for glucose as its metabolic substrate. Advanced, severe DCM is characterized by depletion of adenosine triphosphate (ATP) stores, which may be a consequence of impaired insulin mediated glucose uptake and oxidation at a time when the myocardium prefers glucose as its substrate. We examined the time course and magnitude of myocardial insulin resistance during the evolution of dilated cardiomyopathy. METHODS AND RESULTS: Thirty-four conscious, chronically instrumented dogs were studied at four stages during the evolution of dilated cardiomyopathy (DCM) induced by rapid RV pacing [control, early, late and advanced severe]. Transmyocardial glucose, lactate, and non-esterified fatty acid (NEFA) concentrations were measured in the fasting state. The cellular insulin signaling cascade and ATP levels were measured on myocardial samples. NEFA and insulin concentrations increased early and progressively in DCM in association with increased norepinephrine concentrations and progressive hemodynamic impairment. In advanced DCM but not earlier stages, myocardial ATP levels were decreased by 34%. There was decreased myocardial glucose uptake evident under both basal (-29 +/- 5%) and insulin stimulated (-32 +/- 4%) conditions in advanced, severe DCM, associated with a 31% reduction in GLUT-4 translocation. Importantly, there were no alterations in proximal steps in insulin signaling, but significant reductions in serine (Ser473) phosphorylation of Akt-1. CONCLUSIONS: Advanced, severe DCM is associated with the development of myocardial insulin resistance. There is impaired myocardial glucose uptake and altered myocardial insulin signaling, involving decreased Ser 473 phosphorylation of Akt-1. Myocardial insulin resistance in advanced, severe DCM was also associated with reduced myocardial ATP levels. PMID- 14736547 TI - Sex-specific differences in ventricular expression and function of parathyroid hormone-related peptide. AB - OBJECTIVE: Parathyroid hormone-related peptide (PTHrP) expression is modulated by estrogen. It is expressed in coronary endothelial cells and involved in the endothelium-dependent regulation of coronary resistance and cardiac function. In the present study, we hypothesized that endogenously synthesized and released PTHrP contributes to sex-specific differences in the regulation of cardiac function. METHODS: The influence of sex on ventricular PTHrP expression in normotensive rats was determined via real-time PCR and immunoblot analysis. Sex specific effects of exogenous PTHrP or endogenous released PTHrP were determined in vitro on isolated ventricular cardiomyocytes, Langendorff preparations on isolated hearts and in vivo using different agonistic or antagonistic PTHrP peptides. RESULTS: Ventricular expression of PTHrP was elevated in hearts from female rats compared to male counterparts. Addition of PTHrP(1-36) did not increase left ventricular function in hearts from either sex, but increased coronary flow in hearts from female rats significantly greater than in those from males. 5Ile-PTHrP(1-36), which was used to antagonize endogenously released PTHrP, reduced left ventricular function in females but not males in vitro and in vivo. Under conditions of increased endogenous PTHrP release, i.e. ischemia reperfusion, antagonization of PTHrP significantly reduced post-ischemic recovery in hearts from females but not in those from males. CONCLUSIONS: Sex determines the ventricular expression of PTHrP mRNA and protein. The results indicate that PTHrP may improve cardiac function to a greater extent in women than in men following a brief period of ischemia. PMID- 14736548 TI - Estrogen replacement therapy reverses changes in intramural coronary resistance arteries caused by female sex hormone depletion. AB - OBJECTIVE: We tested the hypothesis that female sex hormone depletion and estradiol replacement therapy significantly influences the biomechanical properties of intramural coronary resistance arteries. DESIGN: Female rats (n=30) were divided into three groups. In group O, rats were subjected to bilateral ovariectomy. Group HRT was subjected to bilateral ovariectomy and estradiol replacement therapy. Rats in group C served as controls. One month after ovariectomy, intramural coronary arteries (approximately 200 microm in diameter) branching from the left anterior descending coronary were isolated, cannulated and studied by microarteriography. Intraluminal pressure was increased in steps between 0 and 90 mm Hg. The steady state diameter at each step was measured. These measurements were repeated in the presence of U46619, a thromboxane (TX) A2 receptor agonist (at a concentration of 10(-6) M), and bradykinin (BK; at 10(-6) M). Finally, Ca2+-free Krebs-induced passive diameter (PD) was measured in each group. RESULTS: Ovariectomy increased spontaneous myogenic tone of coronary arteries (p<0.05), which was normalized by estrogen replacement. Ovariectomy decreased distensibility observed at low pressure, although passive diameter was not changed. Estrogen replacement decreased wall stress and elastic modulus (p<0.05). The thromboxane A2 agonist induced the largest contraction in the ovariectomized group, whereas bradykinin-induced relaxation was the largest in the estrogen replacement group (p<0.05). CONCLUSION: Estradiol hormone replacement therapy (HRT) may exert a beneficial effect on myocardial perfusion in menopause by opposing the deterioration of biomechanical properties of intramural coronary resistance vessels induced by female sex hormone depletion. PMID- 14736549 TI - Effects of chronic heart failure on microvascular oxygen exchange dynamics in muscles of contrasting fiber type. AB - In rat spinotrapezius muscle, chronic heart failure (CHF) speeds microvascular O2 pressure (pO2; index of O2 delivery-to-O2 uptake) dynamics across the rest contractions transition [Cardiovasc. Res. 56 (2002) 479]. Due to the mosaic nature of this muscle, the effect of CHF on microvascular pO2 dynamics in different fiber types remains unclear. OBJECTIVE: Based upon derangements of endothelial function and blood flow responses, we hypothesized that CHF would speed microvascular pO2 dynamics (reduced O2 delivery-to-O2 uptake ratio) in type I muscle (soleus, approximately 84% type I), but not in type II muscle (peroneal, approximately 86% type II [J. Appl. Physiol. 80 (1996) 261]). METHODS: Using phosphorescence quenching, microvascular pO2 was measured at rest and across the rest-contractions transition (1 Hz) in soleus and peroneal of non-infarcted control (control; n=7), and Sprague-Dawley rats with moderate (moderate; elevated left ventricular end-diastolic pressure (LVEDP) 10 +/- 2 mm Hg; n=10) and severe (severe; LVEDP 28 +/- 4 mm Hg; n=5) CHF. RESULTS: The microvascular pO2 mean response time (time delay+time constant) was progressively speeded with increasing severity of CHF in soleus (control, 38.7 +/- 2.0; moderate, 29.1 +/- 1.5; severe, 22.5 +/- 3.9 s; P< or =0.05), but not in peroneal (control=moderate=severe). CONCLUSION: As type I fibers are recruited predominately for moderate intensity exercise, the more rapid lowering of soleus microvascular pO2 in CHF would reduce the blood-muscle O2 driving gradient, exacerbate phosphocreatine and glycogen breakdown, and provide a mechanism for slowed O2 uptake kinetics and premature fatigue in CHF. PMID- 14736550 TI - Effects of chronic PGHS-2 inhibition on PGHS-dependent vasoconstriction in the aged female rat. AB - OBJECTIVE: In menopause, aging and decreased estrogen levels are risk factors contributing to impaired vascular function. Previously, in a young ovariectomized rat model, we demonstrated an increase in prostaglandin H synthase (PGHS) dependent vasoconstriction that could be prevented by estrogen replacement. Subsequently, we found that, with aging, estrogen acts through suppression of the PGHS-2 isoform. HYPOTHESIS: Chronic PGHS-2 inhibition reduces PGHS-dependent vasoconstriction in aging. METHODS: Ovariectomized, aged (12 months) Sprague Dawley rats were treated with a placebo (n=7), or the PGHS-2 inhibitor (NS-398, s.c. 3 mg/kg) for 1 week (n=6) or 4 weeks (n=6). Methacholine (endothelium dependent dilator) and phenylephrine (adrenergic constrictor) were used to assess vascular function in the absence or presence of the nonselective PGHS inhibitor, meclofenamate (1 micromol/l) or the specific PGHS-2 inhibitor NS-398 (10 micromol/l). RESULTS: One week of chronic PGHS-2 inhibition abolished a PGHS dependent shift in methacholine-induced relaxation, while modulation was still observed in phenylephrine constriction. Surprisingly, 4 weeks of PGHS-2 inhibition enhanced PGHS-dependent modulation of vasoconstriction (P<0.05). PGH2/thromboxane inhibition (U-51605, 50 micromol/l) mimicked the results observed with PGHS inhibition among the groups. PGHS-2 expression increased with chronic PGHS-2 inhibition compared to control (P<0.05). CONCLUSIONS: Our data indicate a paradoxical increase in PGHS-dependent vasoconstriction and PGHS-2 expression with prolonged inhibition of PGHS-2 activity. Hence, inhibitors of PGHS-2 activity may not be beneficial in counteracting the vascular dysfunction seen with aging and menopause. PMID- 14736551 TI - Gene transfer of endothelial NO synthase, but not eNOS plus inducible NOS, regressed atherosclerosis in rabbits. AB - The effects of in vivo gene transfer of endothelial nitric oxide synthase (eNOS) and inducible NOS (iNOS) genes on severe atherosclerosis were investigated in rabbits. The recombinant adenoviruses, Ad.eNOS and Ad.iNOS, which respectively express eNOS and iNOS, were constructed. Atherosclerosis was induced by a balloon injury followed by a high cholesterol diet for 12 weeks. The rabbits were divided into six groups: Gp cont (no treatment); Gp null (adenovirus sham-infected); Gp eNOS (Ad.eNOS); Gp iNOS (Ad.iNOS); Gp e+i (Ad.eNOS plus Ad.iNOS); and Gp heNOS (a high dose of Ad.eNOS). Examinations were carried out 7 days after gene transfer. Plasma lipid levels were not significantly changed, but transfection with Ad.eNOS (Gp eNOS and Gp heNOS) decreased the tissue cholesterol concentration and regressed atherosclerotic lesions. Vessels treated with Ad.iNOS (Gp iNOS and Gp e+i) showed iNOS staining in the atheroma, and slight staining at other parts of the vessels; those treated with Ad.eNOS showed eNOS staining in the endothelium and subintima, and slight staining at other parts. Ad.eNOS transfection, but not Ad.iNOS or Ad.eNOS+Ad.iNOS transfection, improved the impaired aortic endothelium dependent relaxation (EDR) and basal NO-dependent response, increased tissue cyclic GMP (cGMP), and decreased the release of O2- from vessels. eNOS treatment showed a decreasing tendency in regions with peroxynitrite staining, MMP1 staining, and suspected apoptosis. In conclusion, in vivo gene transfer of eNOS, but not iNOS or eNOS plus iNOS, regressed atherosclerosis. The relations among NO, O2-, and peroxynitrite may be critical, and lipid resorption from the lesions may be responsible for the regression. PMID- 14736552 TI - Angiotensin-(1-7) modulates vascular resistance and sympathetic neurotransmission in kidneys of spontaneously hypertensive rats. AB - OBJECTIVE: Angiotensin (Ang)-(1-7) generated from Ang I and II is reported to act as an endogenous angiotensin-converting enzyme (ACE) inhibitor and angiotensin type 1 (AT1)-receptor antagonist in vitro and in vivo. Ang-(1-7) has been suggested to play an important role in hypertension. METHODS AND RESULTS: Therefore, we tested whether Ang-(1-7) differentially modulates vascular resistance and neurotransmission in isolated kidneys of spontaneously hypertensive rats stroke prone (SHR-SP) and Wistar-Kyoto rats (WKY). Ang-(1-7) was administered in three concentrations (0.1, 1 and 10 micromol/l) to prevent Ang I- and Ang II-induced pressor responses and facilitation of noradrenaline release. There were indeed concentration-dependent strain differences. Ang-(1-7) prevented Ang I- and Ang II-mediated changes in vascular resistance more potently in SHR-SP than in WKY by inhibiting ACE and by blocking AT1-receptors. Ang-(1-7) by itself had no influence on renal vascular tone in both strains. Ang-(1-7) inhibited Ang I-mediated facilitation of noradrenaline release more potently than Ang II-mediated facilitation of noradrenaline release. Ang-(1-7) by itself enhanced noradrenaline release from SHR-SP, but not from WKY kidneys. CONCLUSION: Ang-(1-7) had a greater impact on Ang I and Ang II modulation of renal vascular resistance in SHR-SP than in normotensive rats. Furthermore, Ang-(1-7) by itself has facilitatory presynaptic effects on noradrenaline release but no postsynaptic effects on renal vascular resistance in SHR-SP. Since plasma levels of Ang-(1-7) accumulate during ACE-inhibitor or AT1-receptor antagonist therapy, Ang-(1-7) could contribute to antihypertensive effects of these agents. PMID- 14736553 TI - Cytosolic prostaglandin E2 synthase/p23 but not apoptosis-linked gene 2 is downregulated in human atherosclerotic plaques. PMID- 14736554 TI - [Coeliac disease: disease of the future?]. PMID- 14736555 TI - [Extra-meningeal meningococcal infection: report of 14 cases]. AB - INTRODUCTION AND METHOD: Fifty-five patients (17 adults, 38 children) with meningococcal infection were admitted between 1986 and 2002 in a university hospital (500 beds). Fourteen of them (nine adults, five children) presented with an extra-meningeal infection. We compared adults and children presentations. RESULTS: All adults had immunodeficiency. Septic locations were various (three bacteriemia, four pneumoniae, one infected ascitis, one cutaneous abscess). All patients received amoxicillin or third generation cephalosporin. Hospitalisation was prolonged (mean: 47 days). Seven patients required intensive care unit admission, and two of them died. All children (all were less than 36-month-old) presented with fever. Only one was immunodeficient (infected by human immunodeficiency virus). Neisseria meningitidis grew from blood in four, and in the throat for the remaining one. Hospitalisation was of short duration (mean: 4 days) and none of the children required intensive care unit. All the children recovered rapidly with antibiotics. CONCLUSION: Outcome of extra-meningeal infection with N. meningitidis is different in adults and children. Adults present with immunodeficiency, infection is severe and patients present with various clinical features; children have a more homogeneous clinical presentation (fever) and outcome is excellent. PMID- 14736556 TI - [MRI of central nervous system in a series of 58 systemic lupus erythematosus (SLE) patients with or without overt neuropsychiatric manifestations]. AB - PURPOSE: Central nervous (CNS) involvement in SLE is common and can be evaluated with MRI. The primary goal of this study was to evaluate with high-field MRI the CNS involvement in a series of SLE patients with or without neuropsychiatric symptoms. The secondary goal was to detect a possible relationship between MRI and clinical or biological parameters in SLE. MATERIALS AND METHODS: We correlated the clinical and biological parameters of 58 patients with a lupus defined according to the American College of Rheumatology criteria, including 30 with neuropsychiatric manifestations with conventional and modern MRI (including diffusion weighted-images, high-resolution 3D T1 weighted-images). The population studied was compared to a group of 18 normal controls. RESULTS: In 69% of cases, MRI demonstrated involvement of the CNS both in asymptomatic patients (64.3%) and in patients with neuropsychiatric manifestations (73.3%): microembolic signals, cerebral infarctions (associated with the anti-phospholipid syndrome), atrophy, basal ganglia involvement, posterior leucoencephalopathy, subcortical calcification or hemosiderin deposits (T2*), dilated perivascular spaces. CONCLUSION: MRI with adapted sequences clearly demonstrated the cerebral involvement in approximately 70% of SLE patients with or without neuropsychiatric symptoms. PMID- 14736557 TI - [Quality of life assessment with the MOS-SF36 in patients with systemic sclerosis]. AB - OBJECTIVE: To investigate the metric properties and the validity of the Medical Outcome Study Short Form 36 (SF-36), a questionnaire to assess the quality of life, in patients with either diffuse or limited systemic sclerosis (SS), and to examine the effect of the disease on quality of life. METHODS: Cross sectional study of 86 patients with a SS (64 diffuse SS, 22 limited SS). Disease severity was assessed by clinical examination, pulmonary functional tests and Health Assessment Questionnaire (HAQ) modified for scleroderma. RESULTS: The SF-36 scores values were lower in diffuse than in limited sclerodermie systemique. The Physical Component Score was worse in patients with than without any clinical involvement. This score increased in relation with the number of clinical involvements. The quality of life of patients with SS was correlated to its functional repercussion. CONCLUSION: The quality of life in SS patients is correlated with the clinical severity of the disease. The use of SF-36 to measure the quality of life is useful for the clinical evaluation of patients with SS. PMID- 14736558 TI - [Celiac disease in adults: new aspects]. AB - INTRODUCTION: Celiac disease also known as gluten-sensitive enteropathy is a complex disorder where genetically susceptible individuals develop in typical cases signs of malabsorption after ingestion of cereals. Malabsorption is due to total or subtotal atrophy of the small intestinal mucosa regressing after adherence to a gluten-free diet. The only effective therapy is life-long strict abstinence from wheat, rye and barley. CURRENT KNOWLEDGE AND KEY POINTS: During the last two decades spectrum of clinical features of adult celiac disease has been modified. About 60% of cases are now revealed by extraintestinal manifestations. Moreover, recent studies that used serological methods revealed existence of both latent and silent celiac sprue. The finding of the high frequency of atypical celiac sprue contributed to underestimation of the true prevalence of celiac disease until now, and explained that celiac disease could be unknown for a long time, with increased risk of nutritional deficiencies and malignancy. Concurrently, the finding that celiac disease is primarily associated to a few HLA class II molecules, and recent advent of tissue transglutaminase as the main auto-antigen eliciting anti-endomysial antibodies allowed to propose new pathogenesis hypothesis and efficient screening tests for celiac disease diagnosis. FUTURE PROSPECTS AND PROJECTS: New epidemiological data concerning adult celiac disease must incite to a more systematic screening in patients with atypical but evocative features, or in at-risk subjects. Introduction of enzyme linked immunosorbent assays (ELISA) for the detection for anti-tissue transglutaminase antibodies allows to make use of a new available and efficient diagnosis parameter, which could constitute the main screening test in the near future. PMID- 14736559 TI - [Heparin-induced thrombocytopenia. Pathogenesis, diagnosis and treatment]. AB - INTRODUCTION: Heparin-induced thrombocytopenia is due to the development of IgG antibodies specific to platelet factor 4. More frequently observed after administration of unfractionated heparin, this complication is characterized by a delayed decrease in platelet count (after the 5th day of treatment) associated with venous and/or arterial thromboses. CURRENT KNOWLEDGE AND KEY POINTS: It is often difficult to discard another potential cause for thrombocytopenia and to demonstrate by Elisa or platelet activation tests the presence of heparin dependent antibodies is therefore mandatory in every patient. Withdrawal of heparin treatment is always necessary as well as the administration of an alternative antithrombotic agent. Danaparoid sodium (mixture of glycosaminoglycanes mainly with anti-Xa activity) or lepirudin (a recombinant hirudin with only antithrombin activity) are both recommended but these two drugs are associated with a significant risk of bleeding in case of renal failure. Oral anticoagulants such as coumadin can only be given when platelet count is normalized and if the clinical evolution is favorable. Potent antiplatelet agents (ilomedine or tirofiban) can be used in specific situations necessitating heparin such as extracorporeal circulation. FUTURE PROSPECTS AND PROJECTS: Early administration of coumadin for the treatment of venous thromboembolic disease efficiently prevent the occurrence of heparin-induced thrombocytopenia, which could disappear in the future with a wider use of the new antithrombotic agents, fondaparinux and ximelagatran. PMID- 14736560 TI - [Antibiotic-associated diarrhea in the elderly]. AB - PURPOSE: Most of the antibiotic-associated diarrhea (AAD) cases result from a transient disturbance in the function of the normal intestinal flora and are spontaneously solved when discontinuing the antibacterial therapy. However, a mild diarrhea lasting several days may induce a dehydration or worsen a denutrition in frail elderly people. CURRENT KNOWLEDGE AND KEY POINTS: The incidence of AAD varies between 5 and 25% depending on the concerned antibiotic. Only 10-20% of all AAD cases are caused by infection, especially with Clostridium difficile, for which advanced age is a major risk factor. The first biological exam to perform when severe AAD or in frail people is the detection of C. difficile toxins, especially in elderly patient treated with beta-lactam antibiotics. Nevertheless, other infectious organisms causing AAD may be considered, as Staphylococcus aureus when predominant in stool cultures from patients treated with fluoroquinolones or as Klebsiella oxytoca when isolated in bloody diarrhea from patients treated with ampicillin. Elevated fecal counts of Candida spp. found in patients treated with antibiotics is rather the consequence of therapy than the cause of AAD. The prevention of AAD is based on a rational antibiotic use to avoid endogenous selection of C. difficile and on the improvement of the hygiene measures to limit the exogenous transmission of the bacteria or related spores by spoiled hands. FUTURE PROSPECTS: Simultaneous prescription of non-pathogenic living organisms, capable of re-establishing the equilibrium of the intestinal flora, should be better described, especially in elderly people, because of its important economic impact. PMID- 14736561 TI - [Helicobacter pylori infection and antimicrobial agents resistance]. AB - PURPOSE: Seven days triple therapies combining a proton pump inhibitor (PPI) and 2 antimicrobial agents (clarithromycin [C], amoxicillin [A], metronidazole [M]), are recommended for the treatment of Helicobacter pylori infection. The eradication failures have increased these last years, particularly in France (about 30%). They are essentially related to the development of antimicrobial agents resistance, mainly concerning macrolides and nitro-imidazoles. CURRENT KNOWLEDGE AND KEY POINTS: Primary resistance to clarithromycin is variable, but reaching now about 10% throughout the world and about 20% in France. It reduces the eradication success rate at 25%. The secondary resistance is very high, contra-indicating the use of clarithromycin in second line regimens. Primary resistance to amoxicillin has recently appeared, but remains very low until now, less than 2%, as the tetracyclin (T) resistance. Primary resistance to metronidazole is 3 times higher than macrolides resistance, but its determination is less accurate. Metronidazole resistance reduces eradication rate of about 25%, leading to the use of metronidazole in second line therapy, in increasing the triple therapy duration at 14 days (PPI-A-M), or in combination with quadruple therapy (Bismuth-PPI-T-M). Other rescue-treatments are efficacious, based on ranitidine bismuth citrate combined regimens or on rifabutine (R) based regimens (PPI-A-R). FUTURE PROSPECTS AND PROJECTS: The recent knowledge of the mutations mainly responsible for H. pylori resistance to antimicrobial agents now allows the development of detection methods based on the study of bacterial DNA. These methods have been validated for clarithromycin and should favour in the near future the determination of resistance by the use of biopsy culture or directly on the gastric biopsy. PMID- 14736562 TI - [Varicose veins. Chronic venous insufficiency. Lower limbs venous ulcers. Point of view]. AB - PURPOSE: Lower limbs chronic venous disorders are still considered as a minor disease from a university hospital point of view, yet it is a very common problem, affecting the quality of life, generating disability, and expensive for healthcare resources. Its teaching is neither satisfactory nor easy to do. KEY POINTS: Apart from the fact that it rarely causes death, the complexity of venous anatomy and physiopathology, the lack of animal model, common practices lumping together visible varicose veins and chronic venous insufficiency, and moreover a certain carelessness in the vocabulary probably largely explain these difficulties. Our purpose is to discuss chronic venous disorders (varicose veins, chronic venous insufficiency, venous ulcers) on the basis of semantic, of anatomical, haemodynamic and clinical forms and finally of nosology. PMID- 14736563 TI - [Systemic lupus erythematosus and/or relapsing polychondritis with catastrophic antiphospholipid syndrome]. AB - INTRODUCTION: Antiphospholipid syndrome is frequently associated with auto-immune disorders. EXEGESIS: We report the case of a 59-years-old woman with an antiphospholipid syndrome associated with systemic lupus erythematosus and/or relapsing polychondritis. CONCLUSION: This fatal evolution as a consequence of a delayed management shows that fast and invasive steps are required for diagnosis and treatment of systemic diseases. PMID- 14736564 TI - [A case of Influenza virus encephalitis in south of France]. AB - INTRODUCTION: Influenza virus outbreaks occur each year, in France, during autumn and winter. Influenza-associated acute encephalitis were reported during epidemics or pandemics. Sporadic cases are rarely identified probably because influenza virus is not searched among etiology of febrile encephalitis. EXEGESIS: We report a case of influenza-associated encephalitis complicated by adrenal insufficiency in a young woman. Diagnosis was based on seroconversion of serum influenza virus A antibodies (complement fixation test). Follow up of the patient showed a total recovery. CONCLUSION: Influenza must be searched for any febrile encephalitis occurring during winter. Reverse transcriptase polymerase chain reaction (RT-PCR) on cerebrospinal fluid should be assessed. It is not actually a routine technique and we do not know yet if it is accurate enough for diagnosis. So, it is important to identify influenza virus and obtain documentary evidence concerning neurological impairment. Nevertheless, a better understanding of pathogenesis and use of vaccination are needed to improve prognosis. PMID- 14736565 TI - [Shrinking lung syndrome and systemic auto-immune disease]. AB - INTRODUCTION: Shrinking lung syndrome usually manifest in dyspnea, decreased lung volume associated with elevated diaphragm. It reports with systemic autoimmune disease and physiopathological mechanism is controversial. EXEGESIS: We report three shrinking lung syndrome observations in which two cases were diagnosed at the time to onset of autoimmune disease. The three patients were treated with corticosteroid, two of them necessitated theophylline. Review of the literature highlight 60 cases and permit to discuss physiopathological mechanisms which remain uncertain. Diaphragmatic dysfunction (because of myositis or neuropathy) represented by abnormal transdiaphragmatic pressures is actually discussed. CONCLUSION: Shrinking lung syndrome is rare but must be considered in patient with autoimmune disease and dyspnea. The diagnosis can be difficult because of clinical, pathological and functional features which are controversial. The optimum treatment is unknown. PMID- 14736566 TI - [Peutz-Jeghers syndrome]. PMID- 14736567 TI - [Light chain-secreting lymphoplasmocytoid lymphoma revealed by retinal vein occlusion]. PMID- 14736568 TI - [Double location of cardiac hydatid cyst: left ventricle and pulmonary artery]. PMID- 14736569 TI - [Common variable immunodeficiency and Evans's syndrome]. PMID- 14736570 TI - ERP old/new effects at different retention intervals in recency discrimination tasks. AB - Recognition memory studies have suggested that event-related brain potentials (ERPs) may tap into several different memory processes. In particular, two ERP components have been hypothesized as related to familiarity (FN400 old/new effect, 300-500 ms, anterior) and recollection processes (parietal old/new effect, 400-800 ms, posterior). The functional significance of the FN400 old/new effect is uncertain because similar old/new differences have been shown to disappear at moderately long retention intervals. The present study investigated the effects of retention interval (34 min, 39 min, or 1 day) on the FN400 and parietal old/new effects in two different recency discrimination tasks. The results suggest that the FN400 old/new effect can be maintained across 1-day retention intervals, so it may index brain processes capable of contributing to long-term memory. PMID- 14736571 TI - Aberrant brain dynamics in schizophrenia: delayed buildup and prolonged decay of the visual steady-state response. AB - In schizophrenia, aberrant brain activity has been reported both during stimulus processing and at rest. Evoked response amplitude is a function of both the number and synchronization of neurons firing in relation to a stimulus. It is at present unclear whether schizophrenia patients have normal synchronization of neural activity in relation to stimulus processing, and whether the amount and time course of synchronization is related to their evoked response amplitudes. EEG brain dynamics in response to visual steady-state stimulation were assessed in 12 schizophrenia and 12 healthy subjects at three stimulation durations (2, 4, and 6 s). Group differences in the visual evoked potential, the visual steady state response, and the local coherence of the visual steady-state response were evaluated over time. Schizophrenia patients had smaller and delayed event-related potentials. Moreover, they had a slower buildup of steady-state amplitude following stimulation onset and a prolonged decrease after stimulation offset. Groups did not differ during mid-segments of steady-state stimulation. Increase in coherence to stimulation onset did not differ between-groups, but coherence decay of the visual steady-state response following stimulus offset was delayed in schizophrenia patients. The initial response to visual stimulation among schizophrenia subjects, therefore, may be reduced in amplitude due to weak signal strength, not poor coordination between distant cortical regions. The prolonged recovery function of schizophrenia patients' visual system may indicate abnormal nonlinearity in neural response. These findings have implications understanding the nature of evoked response differences between schizophrenia and normal groups especially in repetitive stimulus paradigms. PMID- 14736572 TI - P600 related to rule violation in an arithmetic task. AB - The goal of the present paper was to study if a similar neurophysiological process is required for treating violations of both arithmetical rules and linguistic syntactic structures. It has been shown that syntactic violations elicit the P600/syntactic positive shift (SPS) component, reflecting secondary parsing processes or repairing of an incorrect syntactic structure. However, late positivities, similar to the P600/SPS component, are also elicited by other types of violations (e.g. harmonic anomalies or violations in non-linguistic abstract rules), so this component is thought to be an index of detection for any anomaly in rule-governed sequences. We carried out an experiment where violations of arithmetic rules were presented. These violations were evident to a greater or lesser degree (a number very different or very similar to that which correctly completed a series of seven numbers). The type of rule was also manipulated, and increasing and decreasing series were presented. Results showed a late centro parietal positivity related to arithmetic violations, whose amplitude was larger, the more evident the violation presented was, in both addition and subtraction. It is concluded that a similar neurophysiological process could be required for the processing of violations in numerical sequences and in linguistic syntactic structures. When the rule was broken, another component was present for the adding operation: an early negativity peaking between 250- and 300-ms post stimulus. Regarding this negative peak, although some possible explanations are drawn, further research needs to be carried out in order to gather more knowledge about it. PMID- 14736573 TI - Hostility differentiates the brain metabolic effects of nicotine. AB - The brain mechanisms underlying the cause of nicotine dependence are unknown, however, hostility traits are associated with increased susceptibility to nicotine dependence. We used FDG PET to measure brain metabolic response to nicotine administered by patch while the subject performed the Bushman aggression task in 86 high- and low-hostility subjects. Low-hostility trait subjects demonstrated no significant change in brain metabolic response to nicotine. In marked contrast, high-hostility non-smokers subjects demonstrated dramatic metabolic changes to low dose (3.5 mg patch) as did high-hostility smokers to high dose nicotine (21 mg patch) throughout the brain bilaterally (p<0.025). Correlational analyses demonstrated greater metabolic changes in response to nicotine in subjects with greatest hostility trait measures. The observed differences were not a consequence of plasma nicotine or cotinine levels. These metabolic changes were not observed when subjects performed a sustained attentional task (continuous performance task; CPT). Behaviorally, high-hostility subjects had higher ratings of anger, impatience, irritability and nervousness, and lower ratings of happiness, relaxation and curiosity than low-hostility subjects. Smokers had significantly greater scores on impatience and restlessness than non-smokers. This PET study demonstrates a conspicuous lack of the brain metabolic response to nicotine in low-hostility non-smokers in contrast to a dramatic brain response to nicotine in high hostility subjects. This biological difference in brain metabolic response to nicotine between high and low hostility trait subjects may explain differences in susceptibility to nicotine dependence. PMID- 14736574 TI - Discrimination of word stress in early infant perception: electrophysiological evidence. AB - Language acquisition crucially depends on the ability of the child to segment the incoming speech stream. Behavioral evidence supports the hypothesis that infants are sensitive to the rhythmic properties of the language input. We recorded event related potentials (ERPs) to varying stress patterns of two syllable items in adults as well as in 4- and 5-month-old infants using a mismatch negativity (MMN) paradigm. Adult controls displayed a typical MMN to the trochaic item (stress on the first syllable) as well as to the iambic (stress on the second syllable) item. At the age of 4 months, no reliable discrimination response was seen. However, at the age of 5 months, a significant mismatch response (MMR) was observed for the trochaic item, indicating that the trochee, i.e. the most common stress pattern in German, was separated consistently from the iambic item. Hence, the present data demonstrate a clear development between 4 and 5 months with respect to the processing of different stress patterns relevant for word recognition. Moreover, possible contributions of different filter settings to the morphology of the mismatch response in infants are discussed. PMID- 14736575 TI - Dissociating the cortical basis of memory for voices, words and tones. AB - Human speech carries both linguistic content and information about the speaker's identity and affect. While neuroimaging has been used extensively to study verbal memory, there has been little attention to the neural basis of memory for voices. Evidence from studies of aphasia and auditory agnosia suggests that voice memory may rely on anatomically distinct areas in the right temporal and parietal lobes regions, but there is little data on the broader neural systems involved in voice memory. The present study tested the hypothesis that the neural systems involved in voice memory are functionally distinct from the systems involved in word recognition and are primarily located in the right cerebral hemisphere. Subjects performed two-back tasks in which they were required to alternately remember the voices speaking (Voice condition), and the words they produced (Word condition). A tone memory condition was also included, as a non-speech comparison. The contrast between the Voice and Word conditions revealed greater Voice-related effects in left temporal, right frontal and right medial parietal areas, while the Word-related effects appeared in left frontal and bilateral parietal areas. These findings map out a partially right-lateralized fronto-parietal network associated with voice memory, which can be distinguished from predominantly left hemisphere regions associated with verbal working memory. These results provide further evidence that distinct neural systems are associated with the carrier waves of speech and word identity. PMID- 14736576 TI - Brain potentials during semantic and prosodic processing in French. AB - The present experiment was aimed at investigating the on-line processing of semantic and prosodic information. We recorded the Event-Related brain Potentials (ERPs) to semantically and/or prosodically congruous and incongruous sentences that were presented aurally, to study the time course of semantic and prosodic processing, and to determine whether these two processes are independent or interactive. The prosodic mismatch was produced by cross-splicing the beginning of statements with the end of questions, and vice-versa. Subjects had to decide whether the sentences were semantically or prosodically congruous in two different attention conditions. Results showed that a right centro-parietal negative component (N400) was associated with semantic mismatch, and a left temporo-parietal positive component (P800) was associated with prosodic mismatch. Thus, these two electrophysiological markers of semantic and prosodic processing differed in their polarity, latency and scalp distribution. These differences may indicate that the two processes stem from different underlying generators. However, the finding that the P800 elicited by prosodic mismatch was larger when the sentences were semantically incongruous than congruous suggests that the two processes may be interactive. PMID- 14736577 TI - The influence of response-time demands on electrophysiological correlates of successful episodic retrieval. AB - Event-related potentials (ERPs) were acquired in two memory retrieval tasks. In Experiment 1 a 2.5 s response-time limit was imposed at test, while in Experiment 2 there was no explicit upper limit. There were no other structural differences between the two experiments. The response-time manipulation did not influence the accuracy of memory judgements, but resulted in qualitative changes in the ERP old/new effects that were elicited in the two tasks. In Experiment 2, the ERP old/new effects from 700 ms post-stimulus onwards comprised a relatively greater positivity for correct judgements to old items in comparison to correct judgements to new items. In keeping with findings in previous studies, this relative positivity was largest at anterior sites over the right hemisphere. In Experiment 1, by contrast, the ERP old/new effects during the same time window were most prominent at right hemisphere central electrode locations, and comprised a relatively greater positivity for correct judgements to new rather than to old test items. In combination, the findings in the two experiments are consistent with the view that the imposition of different response-time demands results in the engagement of neurally and functionally distinct processes during episodic retrieval. The time course of these distinct ERP old/new effects suggests that different post-retrieval monitoring operations were engaged according to the time available to make memory judgements. PMID- 14736578 TI - Spatial frequency of visual image modulates neural responses in the temporo occipital lobe. An investigation with event-related fMRI. AB - Our visual environment consists of information ranging from coarse to fine patterns with respect to spatial frequency (SF). Neurophysiological studies using experimental animals have shown that there exist specific SF channels in striate and extrastriate visual cortices. In the present study, we used event-related functional magnetic resonance imaging (fMRI) and healthy subjects to investigate whether manipulation of the SF of visual images modulates neural responses in the temporo-occipital lobes. Subjects were scanned while performing "one-back task" with high-pass or low-pass filtered images of a face and house. We demonstrated that visual attention to the stimuli with high SF more specifically involves cortical activation in the left fusiform gyrus and inferior occipital gyrus as compared to that with low SF. High-SF specificity in the left fusiform gyrus was confirmed by voxel-by-voxel comparison of original images with left-right flipped images. There was no low-SF region in the right hemisphere; however, processing of low-SF images may be category-specific in face- and house-related regions. These results may shed light on the neural correlates of behavioral evidence that high-SF stimuli are handled faster and more accurately when presented to the right visual hemifield than to the left counterpart. The present results were also discussed in a viewpoint of local/global processing and functional asymmetry of cerebral hemispheres. PMID- 14736579 TI - Attention to motion enhances processing of both visual and auditory stimuli: an event-related potential study. AB - The present event-related potential (ERP) study investigated whether attending to a particular direction of motion similarly enhances the processing of auditory and visual stimuli. ERPs were recorded while participants perceived horizontally moving visual and auditory stimuli. Attention was manipulated by asking participants to detect an infrequent target stimulus that was of a specified modality (either visual or auditory) and that moved in a specified direction (either leftward or rightward). Stimuli moving in the attended direction elicited ERPs that were more negative than ERPs to stimuli moving in the unattended direction. This difference started around 140 ms post stimulus onset for visual and around 120 ms for auditory stimuli. The auditory effect had a frontal scalp topography, whereas the visual effect was distributed parieto-occipitally. Later parts of the difference waves were maximal at centro-parietal electrodes for both modalities. Crossmodal effects of attention to motion from one modality to the other could not be detected. The results are discussed with regard to hierarchical models of attention. PMID- 14736580 TI - Biomarkers for pain. PMID- 14736581 TI - Interventions to prevent postherpetic neuralgia: cutaneous and percutaneous techniques. PMID- 14736582 TI - Cystatin C in cerebrospinal fluid is not a diagnostic test for pain in humans. AB - A recent subtractive cDNA cloning study in rats demonstrated an unexpected increase in expression of the proteinase inhibitor, cystatin C in the spinal cord during acute peripheral inflammation, suggesting this protein may be involved in the pathogenesis of persistent pain. A subsequent study of 10 women suggested that prolonged labor pain resulted in increased cystatin C concentrations in cerebrospinal fluid, and that this could be used as a biomarker for pain. To confirm and extend these observations, we measured cystatin C concentrations in cerebrospinal fluid in 131 subjects: 30 normal volunteers without pain, 25 women at elective cesarean section without pain, 60 women in labor with severe pain, and 16 patients with chronic neuropathic pain and tactile allodynia. The median cystatin C concentration in normal volunteers, 2.2 microg/ml, was similar to that previously reported by multiple investigators, and cystatin C concentrations were increased in women in labor (3.9 microg/ml). However, contrary to the previous report, cystatin C concentrations in laboring women with pain did not differ from those of pregnant women without pain (3.7 microg/ml). There was no relationship between duration of painful labor and cystatin C concentration. Patients with neuropathic pain had similar cystatin C concentrations (2.4 microg/ml) to controls. Logistic regression analysis indicated that cystatin C concentrations could not be used to reliably predict the presence of pain in either acute or chronic settings. These data suggest that cystatin C concentration in cerebrospinal fluid is an unreliable diagnostic marker for pain in humans. PMID- 14736583 TI - A randomized trial of electronic versus paper pain diaries in children: impact on compliance, accuracy, and acceptability. AB - Electronic diary assessment of pain and disability has become increasingly popular in adult chronic pain research but use of this methodology with children has received limited attention. The aim of this study was to compare two formats of a prospective daily diary (handheld computer=e-diary; paper diary=p-diary) on children's compliance, accuracy, and acceptability ratings. Sixty children, ages 8-16 (M=12.3) with headaches or juvenile idiopathic arthritis, were randomized to receive either e-diaries administered via home visits (n=30) or p-diaries (n=30) handed out during clinic visits for return by mail. Results demonstrated significant mean differences in diary entries completed between groups, with children with e-diaries completing more days (M=6.6) compared to children with p diaries (M=3.8), P<0.001. Diaries returned by children in the p-diary group contained significantly more errors and omissions compared to diaries returned by children in the e-diary group (which contained none), P<0.001. Children rated both diary formats as highly acceptable and easy to use. A significant gender x diary format interaction (P<0.01) was found for compliance where boys demonstrated greater compliance with the e-diary format. Findings demonstrated that the e-diary was feasible to use with children and showed significantly greater compliance and accuracy in diary recording compared to traditional paper diaries in a population of children with recurrent pain. PMID- 14736584 TI - Communicative dimensions of pain catastrophizing: social cueing effects on pain behaviour and coping. AB - The study was designed to assess whether the social context of a pain experience impacted on the relation between catastrophizing and duration of pain behaviour. Based on a communal coping model, the prediction was that the presence of an observer during a pain procedure would differentially influence the display of pain behaviour in high and low catastrophizers. University undergraduates taking part in a cold pressor procedure were randomly assigned to one of two conditions: (1) participant alone (n=30), or (2) observer present (n=34). Analysis of video records revealed that high pain catastrophizers displayed communicative pain behaviours (e.g. facial displays, vocalizations) for a longer duration when an observer was present compared to high pain catastrophizers who were alone during the pain procedure. The duration of pain management behaviours (e.g. holding, rubbing) did not vary significantly as a function of catastrophizing. When the observer was present, high catastrophizers also reported using fewer cognitive coping strategies than low catastrophizers. The pattern of findings suggests that in the presence of an observer, high pain catastrophizers show a propensity to engage in strategies that more effectively communicate their pain, and are less likely to engage in strategies that might minimize pain. Theoretical implications of the findings are discussed. PMID- 14736585 TI - The effects of failing to control pain: an experimental investigation. AB - Chronic pain patients are often confronted with repeated failure to achieve pain relief. The aim of this study was to experimentally investigate the effects of repeated failing attempts to control pain on pain impact (pain intensity, emotional and physiological responses). Perceived control over an electrocutaneous pain stimulus was manipulated between subjects by success or failure feedback on a task by which control over pain could be acquired. In addition, success or failure at the task was manipulated without suggesting a possibility to control pain. It was hypothesized that successful control would lead to lowest pain impact, whereas failure to control pain would lead to even higher pain impact than absent control. Furthermore, it was hypothesized that failure feedback would increase pain impact when compared to success feedback. Results indicated that repeated failure to control pain increased anger and heart rate responses compared to the other conditions, but not pain intensity. It is concluded that persistent efforts to control pain in the face of failure may lead to the maintenance or exacerbation of physiological and emotional responses. PMID- 14736586 TI - The role of neuroticism, pain catastrophizing and pain-related fear in vigilance to pain: a structural equations approach. AB - The present study aimed at clarifying the precise role of pain catastrophizing, pain-related fear and personality dimensions in vigilance to pain and pain severity by means of structural equation modelling. A questionnaire survey was conducted in 122 patients with chronic or recurrent low back pain. Results revealed that pain catastrophizing and pain-related fear mediated the relationship between neuroticism and vigilance to pain. Furthermore, vigilance to pain was found to be associated with heightened pain severity. Finally, we found that neuroticism moderated the relationship between pain severity and catastrophic thinking about pain. The results strongly support the idea that vigilance to pain is dependent upon catastrophic thinking and pain-related fear. Neuroticism is best conceived of as a vulnerability factor; it lowers the threshold at which pain is perceived as threatening, and at which catastrophic thoughts about pain emerge. PMID- 14736587 TI - Comprehensive description of newborn distress behavior in response to acute pain (newborn male circumcision). AB - One of the most difficult challenges still facing researchers and clinicians is assessing pain in the newborn. Behaviors provide one of the most promising avenues for deepening our fundamental understanding of complex phenomenon like newborn pain, and are key to developing descriptive-level knowledge to further newborn pain assessment efforts. In this ethologically based research, we report on the duration and frequency of neonatal distress behavior to seven distinct noxious and non-noxious but distress-provoking events including baseline (diaper change, post-diaper change, application of arm and leg restraints, post application of arm and leg restraints, circumcision, post-circumcision) associated with newborn surgical pain. Approximately 67 min of videotaped data, involving four neonates who had undergone newborn male circumcision, were coded at 1-s intervals (4010 s in total). A reliably established coding scheme was used to code behaviors as they were observed on videotape for the duration of the seven designated events. This led to the identification of (1) 40 distress behaviors as they occurred along the continuum of distress, (2) eight distress behaviors specific to surgery, (3) 11 classes of behaviors occurring within the five sub-phases of circumcision, and (4) a description of 25 distinct post distress behaviors. Findings support the ability to distinguish distress behaviors specific to pain and the ability to detect prolonged distress as well as individual differences in distress-related pain expression. Findings also justify ongoing use of ethological approaches to further newborn pain assessment and to investigate poorly understood topics such as infant self-regulation within the context of pain (pain recovery). PMID- 14736588 TI - Effects of stimulus duration on heat induced pain: the relationship between real time and post-stimulus pain ratings. AB - Pain is a temporally dynamic experience. Yet, in most instances, pain ratings are acquired in a static fashion and frequently require subjects to retrospectively evaluate the pain experience that occurred in a preceding interval of time. In order to determine which components of the real-time experience of pain contribute to static pain ratings, we obtained real-time (dynamic) and post stimulus (static) ratings using a visual analogue scale during various of durations (5-30 s) of noxious thermal stimulation (43-49 degrees C). For both pain-intensity and pain-unpleasantness, real-time ratings revealed that pain adapted when stimulus temperatures were low to moderate and summated when stimulus temperature was high. Regression analyses examining both pain-intensity and pain-unpleasantness revealed that the mean response and the peak response of real-time ratings significantly contributed to post-stimulus ratings, while temporal components such as perceived duration of pain contributed minimally. Additional regression analyses revealed that mean and peak responses of real-time intensity ratings accounted for much of the variability of post-stimulus unpleasantness ratings whereas real-time unpleasantness ratings accounted for somewhat less of the variability of post-stimulus intensity ratings. Taken together, the close relationship between real-time and post-stimulus ratings of pain across stimulus conditions evoking both adaptation and temporal summation further confirms that post-stimulus, retrospective ratings of pain are valid measures of the real-time experience of pain. PMID- 14736589 TI - Local application of the cannabinoid receptor agonist, WIN 55,212-2, to spinal trigeminal nucleus caudalis differentially affects nociceptive and non nociceptive neurons. AB - Cannabinoid receptor agonists produce analgesia for pains of non-cranial origin. However, their effectiveness for craniofacial pains is currently unclear. In the present study, the cannabinoid CB1/CB2 receptor agonist, WIN 55,212-2 (WIN), was bath applied to the brainstem while activity of spinal trigeminal nucleus caudalis (Vc) neurons evoked by transcutaneous electrical stimulation was recorded in isoflurane anesthetized rats. Neurons were characterized using mechanical and electrical stimulation of the face, and were classified as either low-threshold mechanoreceptive (LTM) or wide dynamic range (WDR). LTM neurons responded to light brushing of the receptive field and received only Abeta primary afferent fiber input. WDR neurons showed a graded response to mechanical stimulation, responding maximally to noxious stimuli, and demonstrated both A- and C-fiber evoked activity. In addition, WDR neurons displayed longer latency, C fiber mediated post-discharge (PDC) activity after repetitive stimulation. Local bath application of 2.0 mg/ml WIN significantly reduced PDC activity (3+/-1% control, P<0.01), C-fiber evoked activity (58+/-9% control, P<0.01), and Abeta evoked activity (57+/-10% control, P<0.01) in WDR neurons. In contrast, LTM Abeta fiber evoked activity increased after local administration of WIN (204+/-52% control, P<0.01). SR141716A, a CB1 receptor antagonist, prevented the effects of WIN on WDR PDC and LTM Abeta evoked activity. These results indicate that cannabinoid receptor agonists may be effective agents for craniofacial pain. Furthermore, the particular sensitivity of PDC activity, a measure of neuronal hyperexcitability, to cannabinoid receptor agonists may be relevant to the treatment of persistent craniofacial pain. PMID- 14736590 TI - The clinical relevance of symptom amplification. PMID- 14736593 TI - [Breast cancer and HST: what risk?]. PMID- 14736592 TI - Norepinephrine and pain. PMID- 14736594 TI - [Polycystic ovary syndrome in pubertal period: clinical, biological, metabolic and genetic polymorphism]. AB - Polycystic ovary syndrome (PCOS) is a common cause of hyperandrogenism in adolescent girls. In its complete post menarchal expression, the syndrome is characterized by the association of typical clinical, biological, and ultrasonographic findings. Many factors have contributed to our knowledge of different clinical forms of PCOS in adolescent girls. They are helpful for clarifying misleading situations in a period of life when diagnosis of PCOS implies a treatment for many years and may interfere with gynecological outcome. During the last 3 years, we had the opportunity to manage in our unit 45 adolescent girls with ovarian hyperandrogenism: 32 of them had PCOS and the other 13 functional ovarian hyperandrogenism defined by clinical and biological hyperandrogenism without ultrasonographic abnormality. In this review, we report, from our personal experience as well as from recent literature data, the different clinical expressions of PCOS in the pubertal period: the classical post menarchal form, the exceptional pre menarchal form, the post precocious pubarche and the post precocious puberty forms, the familial expression as well as the dominant metabolic expression. PMID- 14736595 TI - [Transvaginal ultrasound and voiding disorders after TVT procedure]. AB - OBJECTIVE: This study was carried out to evaluate the ultrasonographic findings on patients with voiding disorders after the tension-free vaginal tape procedure (TVT). PATIENTS AND METHOD: Fifty-four women had ultrasonographic evaluation 3 months after TVT procedure. The prolene tape position was evaluated with a complete sagittal view and the width of the tape and the distance between the bladder neck and the tape were measured. In the same time, patients completely cured by TVT (group 1), postoperative dysuria (group 2), de novo urge incontinence (group 3) and ineffective treatment were evaluated with a self questionnaire and physical examination. Statistical analysis was performed using ANOVA test. RESULTS: The distance between the tape and the bladder neck shows a significant difference between group 1 and 2 (13.2 mm compared to 7.6 mm) and between group 1 and 3 (13.2 mm compared to 17.7 mm). The width of the tape is significantly different between group 1 and 2 (8.8 mm compared to 6.6 mm) and between group 1 and 3 (8.8 mm compared to 6.5 mm). The distance between the tape and the bladder neck in group 2, compared with group 3, is significantly shorter (7.6 mm compared to 17.7 mm). There is no difference between the width of the tape between groups 2 and 3 (6.6 mm compared to 6.5 mm). DISCUSSION AND CONCLUSION: These results show a correlation between voiding disorders after TVT procedure and transvaginal ultrasonographic measurement of the distance between the prolene tape and the bladder neck. PMID- 14736596 TI - [Pregnancy and type 2 diabetes: which fetal prognosis?]. AB - OBJECTIVES: The rise in the prevalence of type 2 diabetes in women of childbearing age leads to an increasing number of pregnant women with type 2 diabetes. But published data on fetal outcome are scarce. PATIENTS AND METHODS: In a prospective study from 1999 to 2002, we assessed fetal outcome (preterm delivery, perinatal mortality, congenital malformations) in 20 pregnancies associated with type 2 diabetes and compared the outcome to 40 pregnancies associated with type 1 diabetes. RESULTS: Women with type 2 diabetes are older (32 +/- 5 vs. 27 +/- 5, P = 0.003), more obese (body mass index: 28.3 +/- 4.8 vs. 22.8 +/- 5.5, P < 0.001) than women with type 1 diabetes. Their pregnancy usually is not planned (10% vs. 55%, P < 0.001). HbA1c during organogenesis is above 8% in 46.6% of type 2 vs. 26.4% of type 1 (P < 0.001). Compared with data obtained in the general population, a fivefold increase in preterm delivery (26.3% vs. 4.7%), a sevenfold increase in perinatal mortality (5% vs. 0.7%) and congenital malformations (15.8% vs. 2.2%) are observed. These results are similar to those obtained in type 1. In planned pregnancy, HbA1c during organogenesis is under 7% with no perinatal death and no major congenital malformation. DISCUSSION AND CONCLUSION: Pregnancy complicated by type 2 diabetes is a high-risk one, as much as in type 1 diabetes. Efficient pre-pregnancy care needs to be strongly encouraged in women with type 2 diabetes who also display many risk factors for adverse fetal outcome. PMID- 14736597 TI - [Use of Misoprostol for medical abortion: a trial of the acceptability for home use]. AB - OBJECTIVE: The object of this trial was to appreciate the acceptability of Misoprostol home use in medical abortion. PATIENTS AND METHOD: From 1 January 2001 to 28 February 2002, 120 voluntary patients were included in the study of medical voluntary pregnancy interruption with Misoprostol at home, and 289 patients were using the protocol "Misoprostol at hospital". RESULTS: The method was well tolerated by all patients. DISCUSSION AND CONCLUSION: The protocol "Misoprostol at home" is accepted under certain conditions: strict criteria selection, strict rules of security, good counseling of patients, and finally the availability at the telephone of the gynecologist to respond to any eventuality. PMID- 14736598 TI - [A simple, low-cost and non-invasive method for screening Y microdeletions in infertile men]. AB - OBJECTIVES: Recent investigations showed a high prevalence of Y chromosome microdeletions in men with severely impaired spermatogenesis. Screening for these men is recommended prior to assisted reproduction techniques. The aim of this study was to set up a simple method to detect Y deletion in infertile men. First, we tested the feasibility of cytobrush to collect oral cells as source of DNA. Second, we compared a classic PCR corresponding to European recommendations to the Promega kit. PATIENTS AND METHODS: Seventeen infertile male patients with previously characterized deletions were included in the present study, after fully informed written consent. Both oral cells and blood were used for DNA extraction. A specific DNA extraction protocol was carried out on the buccal cells. The DNAs were tested for Y deletion screening by two different methods. RESULTS: We retrieved between 4 and 10 microg of DNA per brush from buccal cells, allowing several multiplex PCR. The Promega kit detected all the deletions but one: an AZFa deletion was not detected by the two markers of the kit covering this region. In addition, sY130, sY133 and SY153, included in the kit, are not reliable. DISCUSSION AND CONCLUSIONS: Buccal cells represent a convenient substitute for blood in testing for Y microdeletions. Both false negative and false positive results were obtained with Promega Kit. On the opposite, PCR according to the European recommendations allow the accurate detection of Y microdeletion in our 17 cases, at a lower cost. PMID- 14736599 TI - [Breast myoepithelioma. Diagnostic and therapeutic difficulties. A case report]. AB - Breast myoepithelioma is rare. The case of a 61-year-old female is reported here. The tumor was clinically, radiologically and macroscopically demarcated. Diagnosis was based on histological and immunohistochemical studies. At the time of diagnosis the tumor consisted of myoepithelial cells with few cellular atypias and a low mitotic activity. Tumor cells expressed vimentin, actin, S100 protein and cytokeratin. No local or distant metastases were found. The patient had been treated initially with a large excision of tumor. Local recurrences detected one year after initial surgery and showed a similar morphology and immunoreactivity with higher mitotic activity. Mastectomy with axillary dissection was performed. Neither recurrences nor distant metastases detected two years later. Myoepithelial tumors which are generally considered as benign or low-grade lesions can give rise to a wide range of clinical evolution. PMID- 14736600 TI - [Atypical lipomatous tumour of the vulva. About one case]. AB - Liposarcoma of the vulva is a rare entity. This unusual localization with atypical clinical and histological appearance may induce diagnostic and treatment delay. We report the 13th case shown in the literature in a 31-year-old woman initially treated for a vulvar lipoma. Arguments based on clinical short term recurrence, histological infiltrating adipocytes, and cytogenentical findings evoked well-differentiated liposarcoma. Even though cytogenetic abnormalities, involving MDM2 and CDK4 genes, have been found, a certainty in malignity diagnosis could be difficult. In these cases, treatment decision may be uneasy. This case report recalls difficulties encountered in uterine hypercellular leiomyomas. PMID- 14736601 TI - [Sexual function following hysterectomy]. AB - OBJECTIVE: This paper examines the evidence about the impact of hysterectomy on one aspect of quality of life, sexuality. PATIENTS AND METHODS: A systematic review of the French and English language literature was conducted using the search terms "sexuality", "hysterectomy", "libido", and "orgasm". RESULTS: Twenty one studies were found, ten prospective and 11 retrospective. Outcome measures were mostly postoperative libido and orgasm. Most studies did not consider important confounding factors. The majority of authors found either no change or an enhancement of sexuality in women who had an hysterectomy. DISCUSSION AND CONCLUSION: The majority of research evaluating the effect of hysterectomy on sexuality was poorly designed. The available evidence shows that hysterectomy did not adversely affect sexuality. A number of confounding factors should be taken into account in future studies. The role of the gynecologist in the preparation to this kind of operations is very important. PMID- 14736602 TI - [Diagnosis and current concepts of management of advanced abdominal pregnancy]. AB - Authors report a case of abdominal pregnancy diagnosed by MRI at 17 SA with prospective follow-up and planned delivery at 37 SA. The diagnosis is clinically suspected when extra-uterine pregnancy risk factors or history of uterine trauma are present. This is confirmed by MRI, which may be considered as the gold standard. A conservative management may be proposed when the diagnosis is made after 20 weeks and under the following conditions: absence of fetal growth malformation, placental implantation remote from the upper abdomen, good maternal condition, close management in a hospital setting of the patient previously informed of the risks and outcomes. Placental location on the uterus seems to be a major positive factor of outcome for these pregnancies. Materno-fetal follow-up is based on physical examination, repeated ultrasonic investigations with Doppler imaging and daily fetal heart rate monitoring. In the absence of complications, a laparotomy should be planned at 34 weeks. The placenta may not be removed when a serious risk of hemorrhage is feared. PMID- 14736603 TI - [Colposcopy: the value of HPV testing in clinical practice]. AB - The indications for colposcopy have changed recently because of the new Bethesda terminology, the introduction of HPV testing in clinical practice, and the latest consensus guidelines on management of patients with an abnormal cervical cytological test. Colposcopy remains the reference technique to assess patients with abnormal cytological test results, especially those with ASC-H, LSIL, HSIL, and AGC. In women with an ASC-US result, colposcopic examination of only those who test positive for high-risk HPV increases the specificity of the technique. When liquid-based cytology is used, HPV DNA testing is the preferred approach in these women. In primary screening using combined cytology and HPV DNA testing in women over the age of 30, colposcopy is indicated in patients with normal cytology and two HR HPV DNA positive tests performed at a nine month interval. For the follow up of untreated patients with ASC-US/LSIL and CIN I, colposcopy carried out at one year after a single HR HPV DNA positive test is as sensitive as colposcopy after two or three abnormal cytology tests. After excision or conization in patients with high-grade CIN, colposcopy after a single HR HPV DNA positive test is as sensitive as cytology testing and colposcopy at six months. Therapeutic decisions must not be based solely on the results of HPV DNA testing except in specific cases. PMID- 14736604 TI - [Raloxifene and breast: from the SERMs concept to its place in clinical practice]. AB - Raloxifene is a second generation Selective Estrogen Receptor Modulator. It is indicated in prevention and treatment of post-menopausal osteoporosis. This drug has an estrogen agonistic effect on bone tissue, an estrogen-antagonistic effect in breast and endometrial tissue. It inhibits estrogen induced breast cells proliferation and, in animal studies, it prevents the growth of chemically induced mammary tumors. Raloxifene has a limited effectiveness in women with advanced breast cancers. The effects of raloxifene after treated breast cancer and sequential use of tamoxifene after raloxifene (or the contrary) are not known and should be tested in specific pre-clinical and clinical studies. In osteoporotic women included in the MORE trial, the risk reduction of hormone dependent breast cancer due to raloxifene raises the issue of some effectiveness of raloxifene in breast cancer prevention. The STAR trial is currently comparing tamoxifene vs raloxifene in breast cancer prevention in women at increased risk of breast cancer. Like tamoxifene, raloxifene increases the risk of venous thrombo-embolic events. On the other hand, in a subgroup of women at increased vascular risk in the MORE trial, the reduction of cardiovascular events raises the issue of an effect of raloxifene in the prevention of coronary events. The ongoing RUTH trial is testing this assumption and it is comparing raloxifene to a placebo in women at increased vascular risk. Today, in clinical practice, before the results of ongoing trail, raloxifene should be used in the setting of osteoporosis prevention and treatment. PMID- 14736605 TI - [Is the detection of microdeletions of the Y chromosome necessary before ICSI? For the systematic testing of microdeletions of the Y chromosome in infertile men before assisted reproduction procedures]. PMID- 14736606 TI - [The evaluation of the detection of lower urogenital infections by Chlamydia trachomatis in France (Part II) (February 2003)]. PMID- 14736607 TI - [How could the number of abortions in France be reduced? Gynecol Obstet Fertil 2003; 31: 499-503 ]. PMID- 14736608 TI - [Embolization of uterine fibromas: results of 454 cases. Gynecol Obstet Fertil 2003 ; 31: 597-605]. PMID- 14736609 TI - [Embolization of uterine fibromas: results of 454 cases. Gynecol Obstet Fertil 2003; 31: 597-605]. PMID- 14736611 TI - [Management of cervical intraepithelial neoplasm during pregnancy. Gynecol Obstet Fertil 2003; 31: 851-855]. PMID- 14736612 TI - [Should one demonize fertilization in vitro? Gynecol Obstet Fertil 2003; 31: 405 407]. PMID- 14736615 TI - Playground injuries to children. PMID- 14736616 TI - Referral and access to children's health services. PMID- 14736614 TI - The early life origins of asthma and related allergic disorders. PMID- 14736617 TI - Out of hours care. PMID- 14736618 TI - Paediatrics in primary care. PMID- 14736619 TI - The future of primary care paediatrics and child health. PMID- 14736620 TI - The future for child healthcare provision within general practice. PMID- 14736621 TI - Who should provide primary care for children? PMID- 14736622 TI - Nurse practitioners. PMID- 14736623 TI - The future of paediatric primary care and child health. PMID- 14736624 TI - Outcome of non-invasive positive pressure ventilation in paediatric neuromuscular disease. AB - BACKGROUND: Non-invasive positive pressure ventilation (NPPV) has a beneficial effect on nocturnal hypoventilation and hospitalisation rates in adults with static or slowly progressive neuromuscular disease and respiratory failure. Its role in children affected with similar disease processes, however, remains unclear. AIMS: To investigate the impact of NPPV on hospitalisations and sleep related respiratory parameters in children with neuromuscular disease. METHODS: Fifteen children (mean age 11.7, range 3.4-17.8 years) diagnosed with neuromuscular disease who had been started on nocturnal NPPV and had at least one year of follow up since the initiation of such therapy were studied. Patients served as their own controls and comparison was made of the years preceding and following the initiation of NPPV. RESULTS: Children spent 85% fewer days in hospital (mean pre-NPPV 48.0 days, mean post-NPPV 7.0 days) and 68% less days in intensive care after initiation of NPPV (mean pre-NPPV 12.0 days, mean post-NPPV 3.9 days). Sleep study parameters including number of desaturations, apnoea hypopnoea index and transcutaneous pCO2 levels improved after initiation of NPPV. CONCLUSIONS: NPPV can decrease hospitalisations for children with neuromuscular disease and improves sleep related respiratory parameters. A prospective study is now needed to further delineate the role of NPPV in this population of children. PMID- 14736626 TI - The relative efficacy of two brief treatments for sleep problems in young learning disabled (mentally retarded) children: a randomised controlled trial. AB - BACKGROUND: Settling and night waking problems are particularly prevalent, persistent, and generally considered difficult to treat in children with a learning disability, although intervention trials are few. Scarce resources, however, limit access to proven behavioural treatments. AIMS: To investigate the efficacy of a media based brief behavioural treatment of sleep problems in such children by comparing (1) face-to-face delivered treatment versus control and (2) booklet delivered treatment versus controls. METHODS: The parents of 66 severely learning disabled children aged 2-8 years with settling and/or night waking problems took part in a randomised controlled trial with a wait-list control group. Behavioural treatments were presented either conventionally face-to-face or by means of a 14 page easy to read illustrated booklet. A composite sleep disturbance score was derived from sleep diaries kept by parents. RESULTS: Both forms of treatment were almost equally effective compared with controls. Two thirds of children who were taking over 30 minutes to settle five or more times per week and waking at night for over 30 minutes four or more times per week improved on average to having such settling or night waking problems for only a few minutes or only once or twice per week (H = 34.174, df = 2, p<0.001). These improvements were maintained after six months. CONCLUSIONS: Booklet delivered behavioural treatments for sleep problems were as effective as face-to-face treatment for most children in this population. PMID- 14736627 TI - A randomised controlled trial of standing programme on bone mineral density in non-ambulant children with cerebral palsy. AB - BACKGROUND: Severely disabled children with cerebral palsy (CP) are prone to low trauma fractures, which are associated with reduced bone mineral density. AIMS: To determine whether participation in 50% longer periods of standing (in either upright or semi-prone standing frames) would lead to an increase in the vertebral and proximal tibial volumetric trabecular bone mineral density (vTBMD) of non ambulant children with CP. METHODS: A heterogeneous group of 26 pre-pubertal children with CP (14 boys, 12 girls; age 4.3-10.8 years) participated in this randomised controlled trial. Subjects were matched into pairs using baseline vertebral vTBMD standard deviation scores. Children within the pairs were randomly allocated to either intervention (50% increase in the regular standing duration) or control (no increase in the regular standing duration) groups. Pre- and post-trial vertebral and proximal tibial vTBMD was measured by quantitative computed tomography (QCT). RESULTS: The median standing duration was 80.5% (9.5 102%) and 140.6% (108.7-152.2%) of the baseline standing duration in the control group and intervention group respectively. The mean vertebral vTBMD in the intervention group showed an increase of 8.16 mg/cm3 representing a 6% mean increase in vertebral vTBMD. No change was observed in the mean proximal tibial vTBMD. CONCLUSION: A longer period of standing in non-ambulant children with CP improves vertebral but not proximal tibial vTBMD. Such an intervention might reduce the risk of vertebral fractures but is unlikely to reduce the risk of lower limb fractures in children with CP. PMID- 14736628 TI - Return to school after brain injury. AB - AIMS: To examine return to school and classroom performance following traumatic brain injury (TBI). METHODS: This cross-sectional study set in the community comprised a group of 67 school-age children with TBI (35 mild, 13 moderate, 19 severe) and 14 uninjured matched controls. Parents and children were interviewed and children assessed at a mean of 2 years post injury. Teachers reported on academic performance and educational needs. The main measures used were classroom performance, the Children's Memory Scale (CMS), the Wechsler Intelligence Scale for Children-third edition UK (WISC-III) and the Weschler Objective Reading Dimensions (WORD). RESULTS: One third of teachers were unaware of the TBI. On return to school, special arrangements were made for 18 children (27%). Special educational needs were identified for 16 (24%), but only six children (9%) received specialist help. Two thirds of children with TBI had difficulties with school work, half had attention/concentration problems and 26 (39%) had memory problems. Compared to other pupils in the class, one third of children with TBI were performing below average. On the CMS, one third of the severe group were impaired/borderline for immediate and delayed recall of verbal material, and over one quarter were impaired/borderline for general memory. Children in the severe group had a mean full-scale IQ significantly lower than controls. Half the TBI group had a reading age > or =1 year below their chronological age, one third were reading > or =2 years below their chronological age. CONCLUSIONS: Schools rely on parents to inform them about a TBI, and rarely receive information on possible long-term sequelae. At hospital discharge, health professionals should provide schools with information about TBI and possible long-term impairments, so that children returning to school receive appropriate support. PMID- 14736629 TI - Are referrals to occupational therapy for developmental coordination disorder appropriate? AB - AIMS: To assess children referred to the Occupational Therapy Service in Gwent with a presumptive diagnosis of developmental coordination disorder (DCD) in order to investigate the appropriateness of their referral. METHODS: Non-urgent referrals to the occupational therapy team for children with coordination difficulties in Gwent between June 2001 and February 2002 were studied. RESULTS: Eighty nine children, aged 5-10 years, were identified. Thirteen children who would not meet the DCD criteria were excluded. This left 76 children, 67 of whom were actually assessed. If the 15th centile for the Movement ABC is used, 26 children met and 41 failed one of the four criteria in DSM IV (38%). If the 5th centile is used, 21 children met and 46 failed one of the four criteria in the DSM (31%). Of the major groups of referrers, school nurses did the best with 48% success rate; better than the paediatricians with 32%. The worst success rate was in educational psychologists and teachers, with only 20% of cases referred actually having DCD. These differences did not quite reach statistical significance. CONCLUSIONS: Less than a third of referrals to occupational therapy for DCD actually have the diagnosis. This suggests that referrers need further training and guidance. This includes a knowledge and understanding of the DSM IV criteria and their interpretation. This would reduce the number of time consuming, unnecessary assessments being done. A triage procedure with a checklist would be a good way forward and we hope to devise one to assist referrers with this process. PMID- 14736631 TI - Incidence and prevalence of the 22q11 deletion syndrome: a population-based study in Western Sweden. AB - BACKGROUND: Almost all cases of DiGeorge syndrome, velo-cardio-facial syndrome and conotruncal anomaly face syndrome result from a common deletion of chromosome 22q11.2. These syndromes are usually referred to as the 22q11 deletion syndrome (22q11DS), which has a wide phenotypic spectrum and an estimated incidence of one in 4000 births. AIMS: To assess the incidence and prevalence of the 22q11 deletion syndrome in the Western Gotaland Region of western Sweden METHODS: Children below 16 years of age with 22q11DS in a well defined catchment area and population of the Western Gotaland Region were recruited. Diagnosis of 22q11DS was confirmed using a FISH (fluorescence in situ hybridisation) test. Proven 22q11 deletion was the demonstration of one signal in 11 metaphase spreads with fair quality. RESULTS: During the study period in the Western Gotaland Region the mean annual incidence of 22q11DS was 14.1 per 100 000 live births. During the first five years the incidence was 18.1 per 100 000 live births for the whole region and 23.4 per 100 000 live births in Gothenburg, where a multidisciplinary specialist team for 22q11 DS is based. The prevalence was 13.2 per 100 000 children below 16 years of age in the whole region and 23.3 per 100 000 in Gothenburg. CONCLUSION: The number of individuals diagnosed depends on the experience and awareness of the syndrome among specialists who encounter these children and also the severity of the phenotype. The higher frequency of 22q11DS found in Gothenburg is an example of increased awareness. The true incidence and prevalence of this syndrome will only be found through population-based screening, but this would be too expensive and ethically questionable. Screening of specific risk populations would be more justified. PMID- 14736632 TI - Adoption and Children Act 2002. PMID- 14736633 TI - Head injury and limb fracture in modern playgrounds. AB - There were no serious head injuries in modern Cardiff municipal playgrounds with safety surfaces over five years injury surveillance. The literature suggests serious head injuries did occur before the introduction of safety surfaces. PMID- 14736635 TI - The impact of presenting problem based guidelines for children with medical problems in an accident and emergency department. AB - AIMS: To evaluate the impact of presenting problem based guidelines in managing children with either diarrhoea (with or without vomiting) or seizure (with or without fever). METHODS: This prospective observational study with an intervention was based on a paediatric accident and emergency (A&E) department in Nottingham. All patients (either GP or self referred) were acute attenders aged 0 15 years, with a medical presenting problem during 4 months in the spring of 1997 and 1999. Five hundred and thirty-one diarrhoea attendances (292 before guideline implementation and 239 after) and 411 seizure attendances (212 before guideline implementation and 199 after) were recorded. Evidence based and consensus ratified guidelines developed for the study were implemented using care pathway documentation. Process (documentation, time in the department, investigations, treatment) and outcome (admission to hospital, returns to A&E) data were collected from case notes. RESULTS: The percentage of children investigated with blood tests fell significantly (haematology requests in diarrhoea presentations from 11% to 4%, biochemistry in seizure presentations from 29% to 17%). Intravenous infusions in diarrhoea presenters fell (9% to 1%), and more appropriate oral fluids were used. Management time in A&E was reduced (diarrhoea presenters: median of 55-40 minutes, seizure presenters: 80-55 minutes, but remained static for other presenting problems). Marked improvements in documentation were seen. Admission rates for diarrhoea attenders increased (27% to 34%) but remained the same for seizure (69% v 73%). CONCLUSIONS: The implementation of a presenting problem based guideline as a care pathway was associated with improvements in the quality of care by: improved documentation; reduced invasive investigations; more appropriate treatment, and reduced time spent in A&E. PMID- 14736634 TI - Nutrition in the 21st century: what is going wrong. AB - "Things sweet to taste prove in digestion sour" (Shakespeare, Richard II, Act 1, Scene 3, line 236) PMID- 14736636 TI - Patterns of admissions for children with special needs to the paediatric assessment unit. AB - BACKGROUND: Children with special needs present a challenge to those involved in their care. AIMS: To determine the role of the acute assessment unit for these children. METHODS: Case notes and other records were reviewed for information on referrals, admissions, readmission within 7 and 28 days, length of stay, and management of 86 children registered for special needs. The study covered five years between January 1997 and December 2001. RESULTS: Of the 86 children, 48 (58%) were boys; 62 children had cerebral palsy and 52 learning disability. There were 914 episodes, with 44% of these being self referrals and 35% from general practitioners; 35.5% of the episodes were managed in the assessment unit. The average length of stay in hospital was 5 days, ranging from <24 hours to 63 days; 37.5% of those admitted to the ward stayed for less than 24 hours. Respiratory tract infections and seizures were the main reasons for referral and admission. CONCLUSION: Children with special needs tend to have a predictable pattern of conditions requiring inpatient care. One third of the inpatients episodes did not need a prolonged stay in hospital. This latter group of children could be managed at home with support of community nurses. Integrated care pathways need to be developed to minimise disruption to their lives. Appropriate resources should be made available to achieve these goals. PMID- 14736637 TI - Outcome of children with neuromuscular disease admitted to paediatric intensive care. AB - AIMS: To determine the outcome of children with neuromuscular disease (NMD) following admission to a tertiary referral paediatric intensive care (PICU). METHODS: All children with chronic NMD whose first PICU admission was between July 1986 and June 2001 were followed up from their first PICU admission to time of study. The outcomes recorded were death in or outside of PICU, duration of PICU admission, artificial ventilation during admission and following discharge from PICU, and readmission to PICU. RESULTS: Over 15 years, 28 children were admitted on 69 occasions. Sixteen (57%) children had more than one admission. The median duration of PICU admission was 4 days (range 0.5-42). Twenty three per cent of unplanned admissions resulted in the commencement of respiratory support that was continued after discharge from the PICU. Severity of functional impairment was not associated with longer duration of stay or higher PRISM scores. Ten children (36%) died, with four (14%) deaths in the PICU. A higher proportion of children with severe limitation of function were among children that died compared to survivors. CONCLUSION: Most children with NMD admitted to the PICU recover and are discharged without the need for prolonged invasive ventilation. However, in this group of children, the use of non-invasive home based ventilation is common and they are likely to require further PICU admission. PMID- 14736638 TI - Correlation of simultaneously obtained capillary, venous, and arterial blood gases of patients in a paediatric intensive care unit. AB - AIMS: To investigate the correlation of pH, partial pressure of oxygen (PO2), partial pressure of carbon dioxide (PCO2), base excess (BE), and bicarbonate (HCO3) between arterial (ABG), venous (VBG), and capillary (CBG) blood gases. METHODS: Patients admitted to the paediatric intensive care unit (PICU) in Cukurova University between August 2000 and February 2002 were enrolled. RESULTS: A total of 116 simultaneous venous, arterial, and capillary blood samples were obtained from 116 patients (mean age 56.91 months, range 15 days to 160 months). Eight (7%) were neonates. Sixty six (57%) were males. pH, PCO2, BE, and HCO3 were all significantly correlated in ABG, VBG, and CBG. Correlation in PO2 was also significant, but less so. Correlation between pH, PCO2, PO2, BE, and HCO3 was similar in the presence of hypothermia, hyperthermia, and prolonged capillary refilling time. In hypotension, correlation in PO2 between VBG and CBG was similar but disappeared in ABG-VBG and ABG-CBG. CONCLUSIONS: There is a significant correlation in pH, PCO2, PO2, BE, and HCO3 among ABG, VBG, and CBG values, except for a poor correlation in PO2 in the presence of hypotension. Capillary and venous blood gas measurements may be useful alternatives to arterial samples for patients who do not require regular continuous blood pressure recordings and close monitoring of PaO2. We do not recommend CBG and VBG for determining PO2 of ABG. PMID- 14736639 TI - Detection of severe protein-energy malnutrition by nurses in The Gambia. AB - AIM: To test whether nurses can use the WHO integrated management of childhood illness (IMCI) nutrition algorithm to identify reliably severe protein-energy malnutrition in children. METHODS: Nurses were trained to identify severe protein energy malnutrition using IMCI training materials. They identified visible severe wasting and bipedal oedema, and categorised weight-for-age using a growth chart, in consecutive children attending outpatient clinics. Their findings were compared with weight for height Z (WHZ) score, bipedal oedema assessed by a trained observer, and weight-for-age Z score. RESULTS: A total of 352 children were recruited, of whom 34 (9.7%) were severely wasted (WHZ score <-3) and 18 (5.1%) had bipedal oedema. In the detection of severe wasting, the nurses' assessments showed 56% sensitivity, 95% specificity, and 56% positive predictive value (PPV), and for bipedal oedema 22%, 99%, and 57% respectively. Overall, the nurses identified only half of 50 children with severe wasting and/or bipedal oedema and wrongly identified a further 13 children as severely malnourished. Plotting weight for age by the nurses showed 62% sensitivity, 99% specificity, and 89% PPV for the detection of children with very low weight. CONCLUSIONS: Severe malnutrition was both under-diagnosed and wrongly diagnosed by nurses trained in the use of the IMCI nutrition algorithm in a clinic setting in The Gambia. These guidelines for health workers and the training materials, particularly with respect to calculation of age, need further development to improve the detection of malnourished children. PMID- 14736640 TI - The feasibility and acceptability of collecting oral fluid from healthy children for anti-HCV testing. AB - This pilot study investigated the feasibility of surveying, anonymously, HCV infection among healthy children using an oral fluid specimen. Seventy seven per cent of children provided their assent, or where appropriate, consent to participate; 2.8% were anti-HCV positive. Oral fluid collection is acceptable to children and more extensive studies are indicated. PMID- 14736643 TI - Symptomatic vitamin D deficiency among non-Caucasian adolescents living in the United Kingdom. PMID- 14736644 TI - Food allergy in childhood. PMID- 14736641 TI - ESPE/LWPES consensus statement on diabetic ketoacidosis in children and adolescents. AB - Diabetic ketoacidosis (DKA) is the leading cause of morbidity and mortality in children with type 1 diabetes mellitus (TIDM). Mortality is predominantly related to the occurrence of cerebral oedema; only a minority of deaths in DKA are attributed to other causes. Cerebral oedema occurs in about 0.3-1% of all episodes of DKA, and its aetiology, pathophysiology, and ideal method of treatment are poorly understood. There is debate as to whether physicians treating DKA can prevent or predict the occurrence of cerebral oedema, and the appropriate site(s) for children with DKA to be managed. There is agreement that prevention of DKA and reduction of its incidence should be a goal in managing children with diabetes. PMID- 14736647 TI - Prevalence and patterns of occult hip fractures and mimics revealed by MRI. AB - OBJECTIVE: The diagnosis of hip fractures can be difficult on radiography alone. MRI is frequently used to confirm or deny the presence of a minimally displaced hip fracture. This study evaluates the patterns of injury seen on MRI that are difficult to diagnose on radiography. MATERIALS AND METHODS: MRIs of 73 patients who were examined for possible hip fractures and whose radiographic findings were negative or equivocal for hip fracture were reviewed. Seventy-six studies were performed in 73 patients who were between 24 and 102 years old. MRIs were evaluated for the presence and location of bone or soft-tissue injury. Muscle injuries were categorized on the basis of location and type of injury. RESULTS: Forty-six percent (35/76) of the studies showed subtle fractures. Seventeen fractures were in the proximal femur and 18 in the innominate bone. Soft-tissue abnormalities were common, found in 65% of the studies. Twenty percent of the MRI findings were considered normal because there was no apparent finding on the images to explain the patients' symptoms. CONCLUSION: Soft-tissue abnormalities are commonly seen alone or in association with subtle fractures on MRI in the evaluation of patients with a clinical suspicion of hip fracture. MRI is recommended for all symptomatic patients whose radiographic findings are negative for hip fracture. PMID- 14736648 TI - Imaging manifestations of neurosarcoidosis. PMID- 14736649 TI - Underreporting of vertebral fractures on routine chest radiography. AB - OBJECTIVE: Osteoporosis is underdiagnosed and therefore undertreated. We determined the potential usefulness of chest radiography for detecting clinically important vertebral fractures by performing semiquantitative reviews and quantitative digital morphometry on 100 routine chest radiographs taken in the emergency department and comparing the yield of these independent reviews with official radiology reports. MATERIALS AND METHODS: One hundred randomly selected chest radiographs of patients 60 years or older who presented to the emergency department of a tertiary care hospital were evaluated. Radiographs were selected without knowledge of the presenting complaint and were independently reviewed by two board-certified radiologists and a radiology resident. A validated semiquantitative method was used to assess lateral chest radiographs for vertebral fracture. In addition, quantitative digital morphometry was undertaken. A clinically important vertebral fracture was defined as one that was at least moderate to severe (loss of height >or=> 25%). RESULTS: Mean age of the population was 75 years, 47% were women, and 46% were admitted to the hospital. According to the reference radiologist, prevalence of moderate to severe vertebral fractures was 22%. Simple agreement was 87-88% among reviewers; kappa values were moderate (0.56-0.58). The greatest agreement was between the reference standard radiologist and quantitative digital morphometry (89% agreement; kappa = 0.67). Only 55% (12/22) of vertebral fractures we identified were mentioned in the official radiology reports. CONCLUSION: Chest radiography has potential as a screening tool for revealing previously undiagnosed vertebral fractures, although in this study only half of moderate to severe fractures that we identified were mentioned in official reports. PMID- 14736650 TI - Update on the diagnostic radiologist shortage. AB - OBJECTIVE: The purpose of this study is to present the latest information available on the shortage of diagnostic radiologists. MATERIALS AND METHODS: Four sources of information are available, and we present their data: first, the number of jobs for diagnostic radiologists advertised in Radiology and the American Journal of Roentgenology; second, vacancies in academic radiology departments as of July 1, 2003, ascertained by a survey of these departments; third, the ratio of job listings to job seekers at a major professional placement service, the Professional Bureau of the American College of Radiology (ACR); and fourth, diagnostic radiologists' self-reported workload burden, from the ACR's 2003 Survey of Diagnostic Radiologists. RESULTS: Jobs advertised in September November 2003 (latest data available) were 28% fewer than in the same months of 2002. Vacancies per department averaged 3.9 in 2003, compared with 5.4 in 2001 and 5.1 in 2002. Listings per seeker were 1.4 in 2002 (latest data available) compared with 3.0 or more in 1999 and 2000. Responses to a question directly tying changes in workload to changes in income indicated that reported desires for workload reduction and workload increase were approximately equal. CONCLUSION: All four information sources have important limitations, but all indicate that the shortage has considerably eased. We plan to study the causes of this easing and continue to monitor the situation. PMID- 14736651 TI - Improving patient care: the use of a digital teaching file to enhance clinicians' access to the intellectual capital of interdepartmental conferences. AB - OBJECTIVE: We describe a simple method for creating teaching cases from clinical data, radiologic images, surgical images, and images from pathologic slides that are presented at tumor board conferences. CONCLUSION: The resulting interdisciplinary case files are of educational value both during and after conference presentations and can be used by clinicians to gather appropriate historical, laboratory, imaging, surgical, and pathologic data on their patients. This system improves the efficiency and accuracy in gathering patient histories when care is transferred among clinics, the emergency department, and wards. PMID- 14736653 TI - Macroscopic structure of articular cartilage of the tibial plateau: influence of a characteristic matrix architecture on MRI appearance. AB - OBJECTIVE: The purpose of our study was to describe the structural organization of the extracellular matrix of articular cartilage of the tibial plateau and its influence on MRI appearance. MATERIALS AND METHODS: Spin-echo images of 11 resected tibial plateaus acquired at 7 T were compared with the structure of the extracellular matrix as shown by fracture sectioning the samples in the plane of imaging. Four samples were scanned at two different orientations relative to the main magnetic field (B(0)). T2 maps were acquired in two orientations on three of these four samples. RESULTS: On the basis of the presence of reproducible regional variations in the shape of the matrix, a characteristic matrix architecture was described. The location of peak signal intensity and T2 on MRI correlated with the level at which the matrix was estimated to be aligned at approximately 55 degrees to B(0) (r = 0.91). This correlation of matrix orientation relative to B(0) with T2 and signal intensity on MRI was not altered by regional variations in the shape of the matrix or by imaging samples at two different orientations. CONCLUSION: The structure of the extracellular matrix, through its orientation-dependent influence on T2 decay, exerts a strong influence on the MRI appearance of cartilage. At the tibial plateau, a characteristic matrix architecture is associated with an equally characteristic MRI appearance. PMID- 14736654 TI - Treatment of chronic symptomatic vertebral compression fractures with percutaneous vertebroplasty. AB - OBJECTIVE: Most fractures treated with percutaneous vertebroplasty are subacute and less than 1 year old. We report our experience treating chronic vertebral fractures with vertebroplasty. MATERIALS AND METHODS: Our database identified 41 patients with symptomatic fractures more than 1 year old. These patients were categorized into subgroups determined by fracture age: 12 months 1 day-24 months (n = 16) or more than 24 months 1 day (n = 25). Changes in pain and mobility for the study group were compared with those in 49 patients with fractures less than 1 year old. RESULTS: Thirty-three (80%) of the 41 patients in the study group had improvement in pain-seven (17%) had complete and 26 (63%) had partial relief. Forty-five (92%) of the 49 control group patients had improvement in pain-24 (49%) had complete and 21 (43%) had partial relief. The number of patients achieving partial or complete relief of pain was not statistically different between groups (p > 0.05), although complete relief was significantly more frequent in the control group (p = 0.002). Twenty patients (49%) in the study group versus 34 patients (69%) in the control group had improved mobility after vertebroplasty (p = 0.047). Patients with fractures 12 months 1 day-24 months old had improvement in mobility similar to that in patients in the control group (p = 0.962). Fractures more than 24 months 1 day old were associated with significantly less improvement in mobility (p = 0.006). CONCLUSION: Most patients with fractures more than 1 year old will experience clinical benefit from vertebroplasty. Complete relief of pain is more likely when less mature fractures are treated. PMID- 14736655 TI - Radiographic diagnosis of tarsal coalition. AB - OBJECTIVE: The objective of this study was to evaluate the sensitivity of unenhanced radiographic diagnosis of tarsal coalition. MATERIALS AND METHODS: The study had two phases. The first was a retrospective case and control review. Radiographs of 37 feet (15 talocalcaneal coalitions and 15 calcaneonavicular coalitions) and of 17 patients with foot pain and no coalition used as controls were reviewed independently by three observers who had no prior knowledge of the cases. Each observer reviewed the cases for individual signs of coalition and then decided if coalition was present. The second phase of the study was a prospective evaluation by a single observer of 150 consecutive weightbearing foot radiographs obtained to evaluate nontraumatic foot pain. Patients diagnosed as positive for coalition underwent CT. RESULTS: On retrospective review of unenhanced radiographs, observers achieved 100% sensitivity and 88% specificity in the diagnosis of talocalcaneal coalitions. Sensitivity and specificity for calcaneonavicular coalitions ranged from 80% to 100% and 97% to 98%, respectively. Several previously unpublished radiographic signs increased sensitivity of diagnosis. For calcaneonavicular coalition, the new signs were altered navicular morphology and visualization of the bar on the anteroposterior radiograph. For talocalcaneal coalition, the new signs were a dysmorphic sustentaculum tali, nonvisualization of the middle subtalar facet, and shortening of the talar neck. In the prospective phase of the study, three talocalcaneal coalitions were detected with no false-positive results. CONCLUSION: Routine anteroposterior and lateral unenhanced radiographs are a valuable screening tool for tarsal coalition, even when used by inexperienced observers. The newly described signs increase sensitivity of radiographic diagnosis. PMID- 14736656 TI - Arthrography of the shoulder: a simple fluoroscopically guided approach for targeting the rotator cuff interval. AB - OBJECTIVE: The objective of this article is to describe a simplified fluoroscopically guided technique of shoulder joint injection that uses the rotator cuff interval. CONCLUSION: Shoulder arthrography using an anterior approach through the rotator cuff interval is an effective, rapid, and easy-to perform injection technique. In most patients, this technique can be performed using a 1.5-inch (3.8-cm), 22-gauge needle. PMID- 14736657 TI - High-resolution sonography of the triangular fibrocartilage: initial experience and correlation with MRI and arthroscopic findings. AB - OBJECTIVE: The aim of our study was to compare the findings of high-resolution sonography of the triangular fibrocartilage with those of MRI and arthroscopy. SUBJECTS AND METHODS. Thirteen patients with clinically suspected triangular fibrocartilage tears prospectively underwent sonography, followed by MRI, of their wrists. Triangular fibrocartilage tears were classified as predominantly ulnar or predominantly radial. Only the surgeon was aware of the results of both studies, and eight patients subsequently underwent arthroscopy. The findings of the different techniques were compared. RESULTS: For the presence or absence of a tear, seven (87.5%) of eight sonographic examinations correlated with arthroscopy, and 11 (84.6%) of 13 sonographic examinations correlated with MRI. Sonography missed one small radial tear that was detected at arthroscopy and MRI, but sonography showed an ulnar tear in triangular fibrocartilage that appeared normal on MRI. CONCLUSION: High-resolution sonography shows good correlation with MRI and arthroscopy for the evaluation of triangular fibrocartilage tears. Sonography has the potential to be a rapid and cost-effective means of diagnosing tears of the triangular fibrocartilage, particularly those involving the ulnar aspect of the cartilage. PMID- 14736658 TI - Arthrofibrosis associated with total knee arthroplasty: gray-scale and power Doppler sonographic findings. AB - OBJECTIVE: The objective of this study was to determine gray-scale and power Doppler sonographic findings in patients with arthrofibrosis associated with total knee arthroplasty. SUBJECTS AND METHODS. From a consecutive cohort of more than 3000 mobilebearing total knee arthroplasties, 44 cases (1.5%) with arthrofibrosis were identified, of which 38 were recruited for a clinical and sonographic investigation. A control group of 38 patients with a well-functioning total knee arthroplasty was matched. Synovial hypertrophy, presence of neovascularity, patellar tendon thickness, and extent of effusion were assessed. RESULTS: Synovial membrane thickness was significantly (p < 0.001) increased in the arthrofibrosis group (medial, 3.4 mm; lateral, 3.0 mm; suprapatellar, 3.1 mm) when compared with the control group (medial, 2.0 mm; lateral, 2.0 mm; suprapatellar, 1.9 mm). When a cutoff of 3.0 mm was used, sonography had a sensitivity of 84% and a specificity of 82% for detecting arthrofibrosis. Neovascularity (rated as grades 0-3) of the synovial membrane and Hoffa's fat pad was significantly (p 0.054), unenhanced attenuation (p > 0.255), or percentage of the lesion that was exophytic (p > 0.124). CONCLUSION: The attenuation coefficient of a cystic renal lesion increased by no more than 10 H among the unenhanced, corticomedullary, and parenchymal phase scans. PMID- 14736672 TI - Sonography, CT, and MRI of giant cavernous hemangioma of the kidney: correlation with pathologic findings. PMID- 14736673 TI - MRI of an adenomatoid tumor of the tunica albuginea. PMID- 14736675 TI - MDCT in Pancreatic adenocarcinoma: prediction of vascular invasion and resectability using a multiphasic technique with curved planar reformations. AB - OBJECTIVE: The objective of our study was to determine the negative predictive value of MDCT with curved planar reformations for detecting vascular invasion and predicting overall resectability in patients with pancreatic adenocarcinoma. MATERIALS AND METHODS: Imaging findings related to vascular invasion and overall tumor resectability in 25 patients who underwent contrast-enhanced biphasic MDCT evaluation were correlated with actual vessel invasion and overall resectability determined at surgery and pathologic examination. The presence of vascular invasion was assessed in 110 major peripancreatic vessels in 22 patients who underwent resection. RESULTS: On MDCT, 23 (92%) of 25 patients were deemed to have resectable pancreatic adenocarcinoma. The tumors in the remaining two (8%) were considered not resectable because of the presence of vascular invasion (which was confirmed in only one patient at surgery). Of those 23 patients deemed to be candidates for curative resection on the basis of MDCT results, 20 were found to have resectable adenocarcinoma at time of surgery, yielding a negative predictive value for MDCT of 87% (20/23 patients) for overall resectability. In the other three patients, adenocarcinoma was deemed to be unresectable because of small metastases to the liver (two patients) or to the peritoneum (one patient) discovered at surgery. For detection of vascular invasion, MDCT yielded a negative predictive value of 100% (108/108 vessels) with no false-negative findings and an accuracy of 99% (109/110 vessels) with 108 true-negative findings, one true-positive finding, and one false-positive finding. CONCLUSION: Our preliminary data on MDCT show that the technique has excellent negative predictive value for vascular invasion and good negative predictive value for overall tumor resectability in patients with pancreatic adenocarcinoma, suggesting an improvement over previous results reported using single-detector CT. The problem of undetected micrometastases to the liver and peritoneum remains. PMID- 14736676 TI - MDCT of acute lower gastrointestinal bleeding. AB - OBJECTIVE: We evaluated the use of MDCT in the diagnosis and management of lower gastrointestinal bleeding (hematochezia). CONCLUSION: MDCT is proposed as an alternative first-line investigation to locate lower gastrointestinal bleeding before placing the patient under observation or performing embolization or surgery. PMID- 14736677 TI - High-resolution MRI of the anatomy important in total mesorectal excision of the rectum. AB - OBJECTIVE: The surgical removal of a rectal carcinoma and the adjacent lymph nodes in an en bloc package lessens the risk of local recurrence due to residual tumor. Heightened awareness of good surgical techniques has created much interest in the anatomy involved in total mesorectal excision surgery, with particular focus on the fascial planes and nerve plexuses and their relationship to the surgical planes of excision. Clear preoperative depiction of these relationships is of value in determining tumor resectability. The aim of this study was to describe the radiologic appearance of these anatomic structures. SUBJECTS AND METHODS. High-spatial-resolution T2-weighted MRI was performed using a 1.5-T system in cadaveric sections and in patients before they underwent total mesorectal excision surgery. Anatomic dissections of sagitally sectioned hemipelves were compared with MRIs obtained in vivo to establish criteria for visualization of the structures relevant to anterior resection of the rectum. RESULTS: High-spatial-resolution MRI depicted a number of structures of importance in total mesorectal excision surgery. The mesorectal fascia, which forms the boundary of the surgical excision plane in total mesorectal excision, was identified, and the presacral fascia, peritoneal reflection, and Denonvilliers' fascia were also shown. Structures 1-2 mm in diameter were visualized because the contrast resolution afforded by T2-weighted fast spin-echo imaging permitted depiction of the inferior hypogastric nerve plexus and the fascial planes within the posterior pelvis. CONCLUSION: Anatomic landmarks important to the performance of rectal cancer surgery, in particular the mesorectal fascia, may be defined on MRI and are of potential importance in the staging of tumors, assessing resectability, planning surgery, and selecting patients for preoperative neoadjuvant therapy. PMID- 14736678 TI - Peritoneal calcification: causes and distinguishing features on CT. AB - OBJECTIVE: We undertook this study to determine the causes of peritoneal calcification seen on CT and to investigate which CT features distinguish benign from malignant peritoneal calcification. MATERIALS AND METHODS: Seventeen patients with peritoneal calcification were identified through retrospective review of reports from 74765 abdominopelvic CT examinations performed during a 7 year period. We determined the cause of peritoneal calcification by examining medical and histopathologic records. Calcification morphology was classified as nodular or sheetlike on the basis of the consensus interpretation by two independent radiologists. The radiologists also recorded the presence or absence of associated soft-tissue components or lymph node calcification. The association between the CT findings and the cause of calcification was assessed using chi square analysis. RESULTS: Peritoneal calcification was due to peritoneal dialysis (n = 4), prior peritonitis (n = 3), cryptogenic origin (n = 1), or peritoneal spread of ovarian carcinoma (n = 9). Sheet-like calcification was more common in patients with benign calcification (seven of eight patients) than in those with malignant calcification (two of nine patients, p < 0.05). Nodal calcification was seen only in patients with malignant calcification (five of nine patients vs none of eight, p < 0.05). CONCLUSION: Common causes of peritoneal calcification are dialysis, prior peritonitis, or ovarian cancer; sheetlike calcification indicates a benign cause, whereas associated lymph node calcification strongly suggests malignancy. PMID- 14736679 TI - Low mechanical index contrast-enhanced sonographic findings of pyogenic hepatic abscesses. AB - OBJECTIVE: Low mechanical index contrast-specific sonography is a new technique that uses the harmonic capabilities of second-generation contrast agents to produce real-time contrast-enhanced gray-scale images. We describe the contrast specific sonographic findings of pyogenic hepatic abscesses. CONCLUSION: Contrast specific sonography was used to assess eight cases of aspiration-confirmed pyogenic liver abscesses. All cases were correlated with multiphasic helical CT findings. Continuous sonographic exploration allowed recognition of morphologic details not detectable on CT images. Contrast-specific sonograms showed features including rim enhancement, arteries along abscess margins and internal septa, dense and persistent septal enhancement, absent microcirculation in fluid and necrotic components, transient arterial phase hypervascularity around abscesses, and portal phase hypovascularity around abscesses. This constellation of findings is suggestive of liver abscess. PMID- 14736680 TI - Bile leaks after surgery. PMID- 14736681 TI - T2-shortening effect of fibrinogen inclusions on MRI of hepatocellular carcinoma: case report and experimental relaxation measurement. PMID- 14736682 TI - Recurrent abdominal and pelvic abscesses: incidence, results of repeated percutaneous drainage, and underlying causes in 956 drainages. AB - OBJECTIVE: We undertook this study to determine the incidence and results of repeated (secondary) percutaneous abscess drainage performed on recurrent abscesses after successful initial (primary) percutaneous abscess drainage. MATERIALS AND METHODS: Imaging studies from patients who underwent multiple drainages were reviewed to define a cohort of patients who underwent secondary percutaneous abscess drainage after successful initial percutaneous abscess drainage of the same abscess. Medical records of these patients were then reviewed to assess the results of secondary percutaneous abscess drainage. RESULTS: Forty-five abscesses in 43 patients required secondary percutaneous abscess drainage. Twenty-four of the 43 patients avoided surgery. Secondary percutaneous abscess drainage was successful in evacuating the abscess cavity in 39 (91%) of 43 patients. Duration of drainage and time until recurrence were not significant predictors for avoiding surgery. Mean duration of secondary percutaneous abscess drainage was significantly longer than mean duration of primary percutaneous abscess drainage, but duration of secondary percutaneous abscess drainage (25 vs 14 days, respectively; p = 0.007) did not differ significantly between patients who ultimately required surgery and those who did not (17 vs 11 days, respectively; p = 0.10). Time to recurrence ranged from 2 days to 1 year (mean, 51 days). CONCLUSION: After successful primary percutaneous abscess drainage, secondary percutaneous abscess drainage of recurrent abscesses succeeded in evacuating the abscess cavity in most patients, and surgery was avoided by slightly more than half. Patients with postoperative abscesses were significantly more likely to avoid surgery (p = 0.008), whereas patients with pancreatic abscesses were significantly more likely to require it (p = 0.03). PMID- 14736683 TI - Successful interventional treatment of acute internal jugular vein thrombosis. PMID- 14736684 TI - Endovascular treatment of upper extremity septic thrombophlebitis without thrombolysis. PMID- 14736685 TI - MRI before reexcision surgery in patients with breast cancer. AB - OBJECTIVE: The aims of this study were to assess the diagnostic accuracy of MRI in evaluating patients for residual cancer, identify the prevalence of multicentric or multifocal disease, and evaluate the impact of MRI on surgical treatment planning. SUBJECTS AND METHODS. Of 101 potentially eligible patients, 80 candidates for breast conservation therapy who had primary breast cancer in 82 breasts diagnosed by excisional biopsy with close or positive margins were included in the study group. All patients underwent contrast-enhanced MRI before further surgery and subsequently underwent either reexcision lumpectomy or mastectomy with histopathologic correlation. RESULTS: Residual carcinoma, either invasive or in situ, was present in 59.8% of the breasts. The sensitivity and specificity of MRI for detecting residual disease were 61.2% and 69.7%, respectively. Twenty-three additional lesions distant from the biopsy site were identified in 19 breasts, and 18 suspicious lesions underwent biopsy. Histology results indicated that six lesions were malignant, so the overall prevalence was 7.3%. The positive predictive value of identifying an additional suspicious lesion was 33.3%. In 24 breasts, MRI changed which procedure would be performed next from reexcision lumpectomy to mastectomy (n = 9), biopsy of an additional lesion in the ipsilateral (n = 12) or contralateral (n = 2) breast, or neoadjuvant chemotherapy (n = 1). Approximately 25% of the breasts underwent mastectomy as definitive surgical treatment. CONCLUSION: Overlap in the appearances of benign and malignant lesions limits MRI evaluation for residual disease. MRI can show additional suspicious lesions that are likely to be multicentric or multifocal disease. These findings changed the original treatment plan for approximately 30% of breasts. PMID- 14736686 TI - A probabilistic expert system that provides automated mammographic-histologic correlation: initial experience. AB - OBJECTIVE: We sought to determine whether a probabilistic expert system can provide accurate automated imaging-histologic correlations to aid radiologists in assessing the concordance of mammographic findings with the results of imaging guided breast biopsies. MATERIALS AND METHODS: We created a Bayesian network in which Breast Imaging Reporting and Data System (BI-RADS) descriptors are used to convey the level of suspicion of mammographic abnormalities. Our system is a computer model that links BI-RADS descriptors with diseases of the breast using probabilities derived from the literature. Mammographic findings are used to update pretest probabilities (prevalence of disease) into posttest probabilities applying Bayes' theorem. We evaluated the histologic results of 92 consecutive imaging-guided breast biopsies for concordance with the mammographic findings during radiology-pathology review sessions. First, radiologists with no knowledge of the biopsy results chose BI-RADS descriptors for the mammographic findings. After the histologic diagnosis was revealed, the radiologists assessed concordance between the pathologic results and the mammographic findings. We then input the information gathered from these sessions into the Bayesian network to produce an automated mammographic-histologic correlation. RESULTS: We had a sampling error rate of 1.1% (1/92 biopsies). Our expert system was able to integrate pathologic diagnoses and mammographic findings to obtain probabilities of sampling error, thereby enabling us to identify the incorrect pathologic diagnosis with 100% sensitivity while maintaining a specificity of 91%. CONCLUSION: Our probabilistic expert system has the potential to help radiologists in identifying breast biopsy results that are discordant with mammographic findings and discovering cases in which biopsy sampling errors may have occurred. PMID- 14736687 TI - Sonography should not be used for breast cancer screening until its efficacy has been proven scientifically. PMID- 14736688 TI - Prognostic significance of the radiographic pattern of disease in patients with severe acute respiratory syndrome. AB - OBJECTIVE: This study was performed to evaluate the prognostic significance of the radiographic pattern of disease in probable cases of severe acute respiratory syndrome (SARS). MATERIALS AND METHODS: A retrospective review of 439 radiographs was performed for 51 patients with a final diagnosis of probable SARS. Forty-nine patients were followed up for a mean interval of 23 days (range, 2-63 days). RESULTS: Abnormal findings on a chest radiograph were noted at presentation in 80.4% (41/51) of patients. Four radiographic patterns were seen: normal (group 1) in 19.6% (10/51), focal opacity (group 2) in 39.2% (20/51), multifocal opacities (group 3) in 27.5% (14/51), and diffuse air-space opacification (group 4) in 13.7% (7/51). Radiographic progression of disease occurred in 38.8% (19/49) of the patients in groups 1-4. There were no deaths in groups 1 and 2. In group 3, one (7.7%) of the 13 patients died. Five (71.4%) of the seven patients in group 4 died. Overall, 12.2% (6/49) of the patients died, all of whom had diffuse air space opacification on the last chest radiograph. In these patients, medical comorbidity was present in 66.7% (4/6), and the exposure history was known in 83.3% (5/6). Death occurred at a mean interval of 18.2 days (range, 9-36 days) from the initial exposure. CONCLUSION: Patients presenting with normal findings or focal air-space opacity on chest radiographs had a good clinical outcome. Patients with multifocal opacities that progressed to diffuse air-space opacification and patients presenting with diffuse air-space opacification had a high fatality rate, but patients in this group were also older and more likely to have comorbid conditions. Patients with SARS present with recognizable patterns of disease that have prognostic significance. PMID- 14736689 TI - Risk of pulmonary embolism after negative MDCT pulmonary angiography findings. AB - OBJECTIVE: The purpose of our study was to determine the risk of pulmonary embolism in patients who have negative MDCT pulmonary angiography findings. SUBJECTS AND METHODS: In this prospective study, one hundred two consecutive patients with suspected pulmonary embolism underwent MDCT pulmonary angiography. Scans were reviewed jointly by two observers and findings recorded by consensus. Observers noted whether pulmonary embolism or other disease was present. No pulmonary embolism was seen in 85 patients (52 men and 33 women; age range, 20-94 years; mean age, 60 years) who were followed up for a mean of 9 months (range, 4 13 months) for evidence of subsequent pulmonary embolism. RESULTS: One patient had a diagnosis of pulmonary embolism made within 3 weeks of undergoing CT pulmonary angiography. MDCT pulmonary angiography showed additional potentially significant findings in 76% of patients; 47% of these findings were not suspected on chest radiography. CONCLUSION: The risk of pulmonary embolism at a mean of 9 months after negative MDCT pulmonary angiography findings is 1%. In our study of patients without pulmonary embolism, MDCT pulmonary angiography revealed other causes for individual patients' signs or symptoms in most cases. PMID- 14736690 TI - Improved detection of lung nodules on chest radiographs using a commercial computer-aided diagnosis system. AB - OBJECTIVE: The aim of this study was to evaluate the usefulness of a new commercially available computer-aided diagnosis (CAD) system with an automated method of detecting nodules due to lung cancers on chest radiograph. MATERIALS AND METHODS: For patients with cancer, 45 cases with solitary lung nodules up to 25 mm in diameter (nodule size range, 8-25 mm in diameter; mean, 18 mm; median, 20 mm) were used. For healthy patients, 45 cases were selected on the basis of confirmation on chest CT. All chest radiographs were obtained with a computed radiography system. The CAD output images were produced with a newly developed CAD system, which consisted of an image server including CAD software called EpiSight/XR. Eight radiologists (four board-certified radiologists and four radiology residents) participated in observer performance studies and interpreted both the original radiographs and CAD output images using a sequential testing method. The observers' performance was evaluated with receiver operating characteristic analysis. RESULTS: The average area under the curve value increased significantly from 0.924 without to 0.986 with CAD output images. Individually, the use of CAD output images was more beneficial to radiology residents than to board-certified radiologists. CONCLUSION: This CAD system for digital chest radiographs can assist radiologists and has the potential to improve the detection of lung nodules due to lung cancer. PMID- 14736691 TI - Positron emission tomography with FDG to show thymic carcinoid. PMID- 14736692 TI - FDG PET of Rosai-Dorfman disease of the thymus. PMID- 14736693 TI - Coronary MR angiography: comparison of quantitative and qualitative data from four techniques. AB - OBJECTIVE: The optimal coronary MR angiography sequence has yet to be determined. We sought to quantitatively and qualitatively compare four coronary MR angiography sequences. SUBJECTS AND METHODS. Free-breathing coronary MR angiography was performed in 12 patients using four imaging sequences (turbo field-echo, fast spin-echo, balanced fast field-echo, and spiral turbo field echo). Quantitative comparisons, including signal-to-noise ratio, contrast-to noise ratio, vessel diameter, and vessel sharpness, were performed using a semiautomated analysis tool. Accuracy for detection of hemodynamically significant disease (> 50%) was assessed in comparison with radiographic coronary angiography. RESULTS: Signal-to-noise and contrast-to-noise ratios were markedly increased using the spiral (25.7 +/- 5.7 and 15.2 +/- 3.9) and balanced fast field-echo (23.5 +/- 11.7 and 14.4 +/- 8.1) sequences compared with the turbo field-echo (12.5 +/- 2.7 and 8.3 +/- 2.6) sequence (p < 0.05). Vessel diameter was smaller with the spiral sequence (2.6 +/- 0.5 mm) than with the other techniques (turbo field-echo, 3.0 +/- 0.5 mm, p = 0.6; balanced fast field-echo, 3.1 +/- 0.5 mm, p < 0.01; fast spin-echo, 3.1 +/- 0.5 mm, p < 0.01). Vessel sharpness was highest with the balanced fast field-echo sequence (61.6% +/- 8.5% compared with turbo field-echo, 44.0% +/- 6.6%; spiral, 44.7% +/- 6.5%; fast spin echo, 18.4% +/- 6.7%; p < 0.001). The overall accuracies of the sequences were similar (range, 74% for turbo field-echo, 79% for spiral). Scanning time for the fast spin-echo sequences was longest (10.5 +/- 0.6 min), and for the spiral acquisitions was shortest (5.2 +/- 0.3 min). CONCLUSION: Advantages in signal-to noise and contrast-to-noise ratios, vessel sharpness, and the qualitative results appear to favor spiral and balanced fast field-echo coronary MR angiography sequences, although subjective accuracy for the detection of coronary artery disease was similar to that of other sequences. PMID- 14736694 TI - MRI of cardiac morphology and function after percutaneous transluminal septal myocardial ablation for hypertrophic obstructive cardiomyopathy. AB - OBJECTIVE: The objective of our study was to use MRI to show the cardiac morphology and function of patients with hypertrophic obstructive cardiomyopathy after percutaneous transluminal septal myocardial ablation. CONCLUSION: Black blood T2-weighted and contrast-enhanced fast inversion recovery gradient-echo images displayed ablated septal myocardium until 35 weeks after percutaneous transluminal septal myocardial ablation. Central hypointense areas were observed on MRI in patients until 4 weeks after ablation. Black blood and cine steady state free precession MRI were used to assess the decreased septal wall thickness and diameter of the left atrium after ablation as well as the reduced motion of the ablated region. MRI may be useful for evaluation of cardiac structural, signal, and functional changes associated with percutaneous transluminal septal myocardial ablation. PMID- 14736695 TI - CT venography performed with elastic stockings. PMID- 14736696 TI - How should we evaluate CT during arterial portography in patients with cirrhosis? PMID- 14736697 TI - Visualization of hypoattenuation clots on unenhanced CT of the thorax. PMID- 14736698 TI - Pyelovenous Backflow Seen on CT Urography. PMID- 14736699 TI - Magnetic Susceptibility Artifacts on MRI: A Hairy Situation. PMID- 14736700 TI - Peritumoral fatty infiltration of the liver associated with venous drainage from metastatic liver tumor. PMID- 14736701 TI - Peripheral schwannoma lacking enhancement on MRI. PMID- 14736702 TI - Chronic C75 treatment of diet-induced obese mice increases fat oxidation and reduces food intake to reduce adipose mass. AB - Obesity and its attendant disorders, such as type 2 diabetes, are global health problems. We previously reported that C75, an inhibitor of fatty acid synthase (FAS) and stimulator of carnitine palmitoyltransferase I (CPT I), caused anorexia and profound weight loss in lean and genetically obese mice. To approximate human obesity, we utilized a chronic C75 treatment model for diet-induced obese (DIO) mice. Chronic C75 treatment decreased food consumption and increased energy expenditure due to increased fatty acid oxidation in both DIO and lean mice. There was a substantial loss of adipose tissue and resolution of hepatic steatosis in C75-treated DIO mice. Analysis of changes in the expression of hypothalamic neuropeptides demonstrated that the reduced food consumption in C75 treated DIO mice was accompanied by an increase in cocaine and amphetamine related transcript expression but not by changes in neuropeptide Y such as seen with acute C75 treatment of lean mice. Inhibition of FAS and stimulation of CPT I provide a means to achieve stable, sustained weight loss in DIO mice. PMID- 14736703 TI - Restitution of defective glucose-stimulated insulin release of sulfonylurea type 1 receptor knockout mice by acetylcholine. AB - Inhibition of ATP-sensitive K+ (K(ATP)) channels by an increase in the ATP/ADP ratio and the resultant membrane depolarization are considered essential in the process leading to insulin release (IR) from pancreatic beta-cells stimulated by glucose. It is therefore surprising that mice lacking the sulfonylurea type 1 receptor (SUR1-/-) in beta-cells remain euglycemic even though the knockout is expected to cause hypoglycemia. To complicate matters, isolated islets of SUR1-/- mice secrete little insulin in response to high glucose, which extrapolates to hyperglycemia in the intact animal. It remains thus unexplained how euglycemia is maintained. In recognition of the essential role of neural and endocrine regulation of IR, we evaluated the effects of acetylcholine (ACh) and glucagon like peptide-1 (GLP-1) on IR and free intracellular Ca2+ concentration ([Ca2+]i) of freshly isolated or cultured islets of SUR1-/- mice and B6D2F1 controls (SUR1+/+). IBMX, a phosphodiesterase inhibitor, was also used to explore cAMP dependent signaling in IR. Most striking, and in contrast to controls, SUR1-/-) islets are hypersensitive to ACh and IBMX, as demonstrated by a marked increase of IR even in the absence of glucose. The hypersensitivity to ACh was reproduced in control islets by depolarization with the SUR1 inhibitor glyburide. Pretreatment of perifused SUR1-/- islets with ACh or IBMX restored glucose stimulation of IR, an effect expectedly insensitive to diazoxide. The calcium channel blocker verapamil reduced but did not abolish ACh-stimulated IR, supporting a role for intracellular Ca2+ stores in stimulus-secretion coupling. The effect of ACh on IR was greatly potentiated by GLP-1 (10 nM). ACh caused a dose-dependent increase in [Ca2+]i at 0.1-1 microM or biphasic changes (an initial sharp increase in [Ca2+]i followed by a sustained phase of low [Ca2+]i) at 1-100 microM. The latter effects were observed in substrate-free medium or in the presence of 16.7 mM glucose. We conclude that SUR1 deletion depolarizes the beta-cells and markedly elevates basal [Ca2+]i. Elevated [Ca2+]i in turn sensitizes the beta-cells to the secretory effects of ACh and IBMX. Priming by the combination of high [Ca2+]i, ACh, and GLP-1 restores the defective glucose responsiveness, precluding the development of diabetes but not effectively enough to cause hyperinsulinemic hypoglycemia. PMID- 14736704 TI - Novel regulation by Rac1 of glucose- and forskolin-induced insulin secretion in INS-1 beta-cells. AB - Stimulation of insulin secretion by glucose and other secretagogues from pancreatic islet beta-cells is mediated by multiple signaling pathways. Rac1 is a member of Rho family GTPases regulating cytoskeletal organization, and recent evidence also implicates Rac1 in exocytotic processes. Herein, we report that exposure of insulin-secreting (INS) cells to stimulatory glucose concentrations caused translocation of Rac1 from cytosol to the membrane fraction (including the plasmalemma), an indication of Rac1 activation. Furthermore, glucose stimulation increased Rac1 GTPase activity. Time course study indicates that such an effect is demonstrable only after 15 min stimulation with glucose. Expression of a dominant-negative Rac1 mutant (N17Rac1) abolished glucose-induced translocation of Rac1 and significantly inhibited insulin secretion stimulated by glucose and forskolin. This inhibitory effect on glucose-stimulated insulin secretion was more apparent in the late phase of secretion. However, N17Rac1 expression did not significantly affect insulin secretion induced by high K+. INS-1 cells expressing N17Rac1 also displayed significant morphological changes and disappearance of F actin structures. Expression of wild-type Rac1 or a constitutively active Rac1 mutant (V12Rac1) did not significantly affect either the stimulated insulin secretion or basal release, suggesting that Rac1 activation is essential, but not sufficient, for evoking secretory process. These data suggest, for the first time, that Rac1 may be involved in glucose- and forskolin-stimulated insulin secretion, possibly at the level of recruitment of secretory granules through actin cytoskeletal network reorganization. PMID- 14736705 TI - Effects of short- and medium-term calorie restriction on muscle mitochondrial proton leak and reactive oxygen species production. AB - Reductions in cellular oxygen consumption (Vo2) and reactive oxygen species (ROS) production have been proposed as mechanisms underlying the anti-aging effects of calorie restriction (CR). Mitochondria are a cell's greatest "sink" for oxygen and also its primary source of ROS. The mitochondrial proton leak pathway is responsible for 20-30% of Vo2 in resting cells. We hypothesized that CR leads to decreased proton leak with consequential decreases in Vo2, ROS production, and cellular damage. Here, we report the effects of short-term (2-wk, 2-mo) and medium-term (6-mo) CR (40%) on rat muscle mitochondrial proton leak, ROS production, and whole animal Vo2. Whole body Vo2 decreased with CR at all time points, whereas mass-adjusted Vo2 was normal until the 6-mo time point, when it was 40% lower in CR compared with control rats. At all time points, maximal leak dependent Vo2 was lower in CR rats compared with controls. Proton leak kinetics indicated that mechanisms of adaptation to CR were different between short- and medium-term treatments, with the former leading to decreases in protonmotive force (Deltap) and state 4 Vo2 and the latter to increases in Deltap and decreases in state 4 Vo2. Results from metabolic control analyses of oxidative phosphorylation are consistent with the idea that short- and medium-term responses are distinct. Mitochondrial H2O2 production was lower in all three CR groups compared with controls. Overall, this study details the rapid effects of short- and medium-term CR on proton leak, ROS production, and metabolic control of oxidative phosphorylation. Results indicate that a reduction in mitochondrial Vo2 and ROS production may be a mechanism for the actions of CR. PMID- 14736706 TI - Exaggerated 17-hydroxyprogesterone response to intravenous infusions of recombinant human LH in women with polycystic ovary syndrome. AB - Studies using pharmacological gonadotropin stimulation suggest that ovarian steroidogenesis is abnormal in the polycystic ovary syndrome (PCOS). We assessed ovarian steroid secretion in response to near-physiological gonadotropin stimuli in 12 ovulatory controls and 7 women with PCOS. A gonadotropin-releasing hormone receptor antagonist (ganirelix, 2 mg sc) was given to block endogenous LH secretion, followed by dexamethasone (0.75 mg orally) to suppress adrenal androgen secretion. After ganirelix injection (12 h), intravenous infusions of recombinant human LH (0, 10, 30, 100, and 300 IU; each over 8 min) were administered at 4-h intervals in a pseudorandomized (highest dose last) manner. Plasma LH, 17-hydroxyprogesterone (17-OHP), androstenedione, and testosterone were measured concurrently. LH dose-steroid response relationships (mean sex steroid concentration vs. mean LH concentration over 4 h postinfusion) were examined for each subject. Linear regression of 17-OHP on LH yielded a higher (mean +/- SE) slope in PCOS (0.028 +/- 0.010 vs. 0.005 +/- 0.005, P < 0.05), whereas extrapolated 17-OHP at zero LH was similar. The slopes of other regressions did not differ from zero in either PCOS or controls. We conclude that near-physiological LH stimulation drives heightened 17-OHP secretion in patients with PCOS, suggesting abnormalities of early steps of ovarian steroidogenesis. With the exception of 17-OHP response in PCOS, no acute LH dose-ovarian steroid responses were observed in controls or PCOS. Defining the precise mechanistic basis of heightened precursor responsiveness to LH in PCOS will require further clinical investigation. PMID- 14736707 TI - Mitochondrial localization of ERalpha and ERbeta in human MCF7 cells. AB - We observed previously that estrogen treatment increased the transcript levels of several mitochondrial DNA (mtDNA)-encoded genes for mitochondrial respiratory chain (MRC) proteins and MRC activity in rat hepatocytes and human Hep G2 cells. Others have reported detection of estrogen receptors (ER), ERalpha and ERbeta, in mitochondria of rabbit ovarian and uterine tissue. In this study, we have extended these observations. Using cellular fractionation and Western blot with ERalpha- and ERbeta-specific antibodies, we observed that ERalpha and ERbeta are present in mitochondria of human MCF7 cells and that the mitochondrial ERalpha and ERbeta account for 10 and 18%, respectively, of total cellular ERalpha and ERbeta in 17beta-estradiol (E(2))-treated MCF7 cells. We also found that E(2) significantly enhanced the amounts of mitochondrial ERalpha and ERbeta in a time- and concentration-dependent manner and that these effects are accompanied by a significant increase in the transcript levels of mtDNA-encoded genes, i.e., cytochrome c oxidase subunits I and II. Moreover, we demonstrated that these E(2) mediated effects were inhibited by the pure ER antagonist, ICI-182780, indicating the involvement of ERs. Using immunohistochemistry with confocal microscopy and immunogold electron microscopy, we demonstrated that ERalpha and ERbeta are located within the MCF7 cell mitochondrial matrix. Computer analysis identified a putative internal mitochondrial targeting peptide signal within human ERbeta, suggesting an inherent potential for ERbeta to enter mitochondria. These findings confirm the observations of others and provide additional support for this novel localization of the ERs and for a potentially important role of the ER in the regulation of mtDNA transcription. PMID- 14736708 TI - Transplacental delivery of retinoid: the role of retinol-binding protein and lipoprotein retinyl ester. AB - Retinoids are required for normal embryonic development. Both embryonic retinoid deficiency and excess result in congenital malformations. There is little understanding of the physiology underlying retinoid transfer from the maternal circulation to the embryo. We now report studies that explore this process using retinol-binding protein-deficient (RBP-/-) mice and mice that express human RBP on the RBP-/-) background. Our studies establish that dietary retinoid, bound to lipoproteins, can serve as an important source for meeting tissue retinoid requirements during embryogenesis. Indeed, retinyl ester concentrations in the circulations of pregnant RBP-/- mice are significantly elevated over those observed in wild-type mice, suggesting that lipoprotein retinyl esters may compensate for the absence of retinol-RBP during pregnancy. We also demonstrate, contrary to earlier proposals, that maternal RBP does not cross the placenta and cannot enter the fetal circulation. Overall, our data indicate that both retinol RBP and retinyl esters bound to lipoproteins are able to provide sufficient retinoid to the embryo to allow for normal embryonic development. PMID- 14736709 TI - Volume changes and whole cell membrane currents activated during gradual osmolarity decrease in C6 glioma cells: contribution of two types of K+ channels. AB - Volume changes and whole cell ionic currents activated by gradual osmolarity reductions (GOR) of 1.8 mosM/min were characterized in C6 glioma cells. Cells swell less in GOR than after sudden osmolarity reductions (SOR), the extent of swelling being partly Ca(2+) dependent. In nominally Ca(2+)-free conditions, GOR activated predominantly whole cell outward currents. Cells depolarized from the initial -79 mV to a steady state of -54 mV reached at 18% osmolarity reduction [hyposmolarity of -18% (H-18%)]. Recordings of Cl(-) and K(+) currents showed activation at H-3% of an outwardly rectifying Cl(-) current, with conductance of 1.6 nS, sensitive to niflumic acid and 5-nitro-2-(3-phenylpropylamino)benzoic acid, followed at H-18% by an outwardly rectifying K(+) current with conductance of 4.1 nS, inhibited by clofilium but insensitive to the typical K(+) channel blockers. With 200 nM Ca(2+) in the patch pipette, whole cell currents activated at H-3% and at H-13% cells depolarized from -77 to -63 mV. A K(+) current activated at H-1%, showing a rapid increase in conductance, suppressed by charybdotoxin and insensitive to clofilium. These results show the operation of two different K(+) channels in response to GOR in the same cell type, activated by Ca(2+) and osmolarity and with different osmolarity activation thresholds. Taurine and glutamate efflux, monitored by labeled tracers, showed delayed osmolarity thresholds of H-39 and H-33%, respectively. This observation clearly separates the Cl(-) and amino acid osmosensitive pathways. The delayed amino acid efflux may contribute to counteract swelling at more stringent osmolarity reductions. PMID- 14736710 TI - Regulation of the human NBC3 Na+/HCO3- cotransporter by carbonic anhydrase II and PKA. AB - Human NBC3 is an electroneutral Na(+)/HCO(3)(-) cotransporter expressed in heart, skeletal muscle, and kidney in which it plays an important role in HCO(3)(-) metabolism. Cytosolic enzyme carbonic anhydrase II (CAII) catalyzes the reaction CO(2) + H(2)O left arrow over right arrow HCO(3)(-) + H(+) in many tissues. We investigated whether NBC3, like some Cl(-)/HCO(3)(-) exchange proteins, could bind CAII and whether PKA could regulate NBC3 activity through modulation of CAII binding. CAII bound the COOH-terminal domain of NBC3 (NBC3Ct) with K(d) = 101 nM; the interaction was stronger at acid pH. Cotransfection of HEK-293 cells with NBC3 and CAII recruited CAII to the plasma membrane. Mutagenesis of consensus CAII binding sites revealed that the D1135-D1136 region of NBC3 is essential for CAII/NBC3 interaction and for optimal function, because the NBC3 D1135N/D1136N retained only 29 +/- 22% of wild-type activity. Coexpression of the functionally dominant-negative CAII mutant V143Y with NBC3 or addition of 100 microM 8 bromoadenosine to NBC3 transfected cells reduced intracellular pH (pH(i)) recovery rate by 31 +/- 3, or 38 +/- 7%, respectively, relative to untreated NBC3 transfected cells. The effects were additive, together decreasing the pH(i) recovery rate by 69 +/- 12%, suggesting that PKA reduces transport activity by a mechanism independently of CAII. Measurements of PKA-dependent phosphorylation by mass spectroscopy and labeling with [gamma-(32)P]ATP showed that NBC3Ct was not a PKA substrate. These results demonstrate that NBC3 and CAII interact to maximize the HCO(3)(-) transport rate. Although PKA decreased NBC3 transport activity, it did so independently of the NBC3/CAII interaction and did not involve phosphorylation of NBC3Ct. PMID- 14736711 TI - Selective activation of STAT3 in human monocytes stimulated by G-CSF: implication in inhibition of LPS-induced TNF-alpha production. AB - Lipopolysaccharide (LPS) induced tumor necrosis factor (TNF)-alpha production in human monocytes, which was dependent on activation of extracellular signal regulated kinase (ERK), p38, c-Jun NH(2)-terminal kinase (JNK), and nuclear factor (NF)-kappa B. LPS-induced TNF-alpha production was inhibited by granulocyte colony-stimulating factor (G-CSF) and interleukin (IL)-10. G-CSF, like IL-10, exerted the inhibitory effect even when simultaneously added with LPS. Among the signaling pathways, signal transducer and activator of transcription 3 (STAT3) was selectively activated in monocytes stimulated by G CSF or IL-10. G-CSF-mediated inhibition of LPS-induced TNF-alpha production as well as G-CSF-induced STAT3 phosphorylation and suppressor of cytokine signaling 3 mRNA expression were prevented by pretreatment of monocytes with AG-490, an inhibitor of Janus kinase 2. G-CSF did not affect LPS-induced activation of ERK, p38, JNK, and NF-kappa B, indicating that G-CSF affects the pathway downstream or independently of these signaling molecules. G-CSF-induced, but not IL-10-induced, STAT3 phosphorylation was attenuated in the presence of LPS. These findings suggest that G-CSF, like IL-10, inhibits LPS-induced TNF-alpha production in human monocytes through selective activation of STAT3, and the immunomodulation observed in vivo by G-CSF administration may be partly ascribed to the direct effect of G-CSF on monocyte functions. PMID- 14736712 TI - Properties of a tonically active, sodium-permeable current in mouse urinary bladder smooth muscle. AB - Urinary bladder smooth muscle (UBSM) elicits depolarizing action potentials, which underlie contractile events of the urinary bladder. The resting membrane potential of UBSM is approximately -40 mV and is critical for action potential generation, with hyperpolarization reducing action potential frequency. We hypothesized that a tonic, depolarizing conductance was present in UBSM, functioning to maintain the membrane potential significantly positive to the equilibrium potential for K(+) (E(K); -85 mV) and thereby facilitate action potentials. Under conditions eliminating the contribution of K(+) and voltage dependent Ca(2+) channels, and with a clear separation of cation- and Cl(-) selective conductances, we identified a novel background conductance (I(cat)) in mouse UBSM cells. I(cat) was mediated predominantly by the influx of Na(+), although a small inward Ca(2+) current was detectable with Ca(2+) as the sole cation in the bathing solution. Extracellular Ca(2+), Mg(2+), and Gd(3+) blocked I(cat) in a voltage-dependent manner, with K(i) values at -40 mV of 115, 133, and 1.3 microM, respectively. Although UBSM I(cat) is extensively blocked by physiological extracellular Ca(2+) and Mg(2+), a tonic, depolarizing I(cat) was detected at -40 mV. In addition, inhibition of I(cat) demonstrated a hyperpolarization of the UBSM membrane potential and decreased the amplitude of phasic contractions of isolated UBSM strips. We suggest that I(cat) contributes tonically to the depolarization of the UBSM resting membrane potential, facilitating action potential generation and thereby a maintenance of urinary bladder tone. PMID- 14736713 TI - Severe respiratory compromise secondary to cervical disk herniation in two dogs. AB - Two dogs presented with acute tetraparesis, hypoventilation, and bradycardia with a second-degree atrioventricular heart block. Neurological examination localized both lesions to the cervical spine. Diagnostic imaging revealed a ventral extradural compression at the second to third cervical (C(2)-C(3)) region in one dog and at the third to fourth cervical (C(3)-C(4)) region in the other. Following surgical correction of the extruded disk, the hypoventilation and bradycardia resolved. Cervical disk extrusions are a common cause of acute tetraparesis in the dog. This report shows that respiratory and cardiac complications may occur concurrently. The authors recommend screening dogs with cervical myelopathies for respiratory and cardiac dysfunctions and treating appropriately. Prompt surgical intervention and supportive care can improve the prognosis. PMID- 14736714 TI - Acute nontraumatic hemoabdomen in the dog: a retrospective analysis of 39 cases (1987-2001). AB - The medical records of 39 dogs with acute nontraumatic hemoabdomen were identified and reviewed. Anemia and hypoalbuminemia were identified in 36/37 (97%) and 25/33 (76%) dogs, respectively. Coagulopathies were identified in 26/31 (84%) dogs. When a definitive diagnosis was obtained, malignant neoplasia was diagnosed most frequently and occurred in 24/30 (80%) dogs. Hemangiosarcoma accounted for 21/30 (70%) diagnoses. Sixteen dogs underwent exploratory laparotomy, of which seven (44%) survived the perioperative period. Of the dogs that did not undergo surgery, 9/23 (39%) survived to be discharged from the hospital. PMID- 14736715 TI - Packed red blood cell transfusions in dogs with gastrointestinal hemorrhage: 55 cases (1999-2001). AB - Fifty-five dogs received packed red blood cell (PRBC) transfusions for gastrointestinal (GI) hemorrhage during a 26-month period (1999 to 2001), accounting for 11.7% of the PRBC transfusions in that time. Thirty-nine (61%) dogs had an intestinal pathology (primary or secondary) as the cause of GI hemorrhage, including intestinal masses, gastroenteritis, hepatic disease, and renal disease. Nonsteroidal and steroidal anti-inflammatory drug use was found frequently in dogs with GI hemorrhage. Sixteen (39%) dogs were identified as having immune-mediated thrombocytopenia (IMT) and associated GI hemorrhage. Dogs with IMT received more transfusions of PRBC than nonIMT dogs (P<0.03) and received a significantly larger total volume of PRBC (P<0.01) during hospitalization. PMID- 14736716 TI - Septic pericarditis, aortic endarteritis, and osteomyelitis in a dog. AB - A 7-year-old, female spayed rottweiler was referred with a history of an acute onset of collapse attributable to cardiac tamponade. Thoracic radiographs revealed an enlarged cardiac silhouette compatible with pericardial effusion, sternal osteomyelitis, and an unusual mineralized lesion determined later to be within the aortic wall. The pericardial effusion was a septic exudate secondary to infection with Staphylococcus species and hemorrhage into the pericardium through a mineralized aortic lesion. The case demonstrates the importance of complete evaluation of thoracic radiographs in a patient with cardiac disease and the potential value of cytopathological evaluation of pericardial fluid. PMID- 14736717 TI - Prevalence and risk factors for heartworm infection in cats from northern Florida. AB - Necropsies were performed on 630 adult cats in northern Florida to determine the prevalence and risk factors for heartworm infection in cats of this region. Heartworms were identified in 4.9% of cats, and serological evidence of heartworm exposure was present in 17% of cats. Not all cats from which heartworms were recovered were seropositive for heartworm antigen or antibody. There was no association between heartworm infection and co-infection with feline leukemia virus (FeLV) or feline immunodeficiency virus (FIV). Male cats were at higher risk of infection with heartworm, FeLV, or FIV than were females. Because even a single heartworm can cause clinical disease or death in cats, the authors conclude that cats in this region should receive heartworm prophylaxis to prevent heartworm infection. PMID- 14736718 TI - Septicemic salmonellosis in two cats fed a raw-meat diet. AB - Salmonella gastroenteritis and septicemia were diagnosed in two cats presented for necropsy. Both cats resided in the same household and were fed a home prepared, raw meat-based diet. Salmonella was isolated from multiple organs in both cats and from samples of raw beef incorporated into the diet fed to one of the cats. Subtyping of the bacterial isolates yielded Salmonella newport from one cat and from the diet it had been fed. This report provides evidence that the practice of feeding raw meat-based diets to domestic cats may result in clinical salmonellosis. PMID- 14736719 TI - Cutaneous paecilomycosis in a cat. AB - A cat was presented for a 2-year history of a recurrent, soft-tissue swelling of the left metacarpal region. The mass was excised and submitted for aerobic and anaerobic bacterial culture, fungal culture, and histopathological examination. Cultures revealed the organism Paecilomyces lilacinus, and histopathological examination showed a nodular mycotic granuloma. Itraconazole (10 mg/kg body weight, per os [PO], q 24 hours) was administered and continued for a total of 60 days, with a swelling of the upper lip occurring 3 months after the initial presentation. Subsequent surgical excisions and debridements along with treatment with itraconazole (20 mg/kg body weight, PO, q 24 hours) for a total of 4 months were curative. PMID- 14736720 TI - Clinical evaluation of rabies virus meningoencephalomyelitis in a dog. AB - A 6-month-old, female, mixed-breed dog presented for acute, progressive, flaccid paraplegia and bilateral pelvic-limb hyperesthesia. A lymphocytic pleocytosis with 366 mg/dL protein was found on cerebral spinal fluid (CSF) evaluation. Electromyography (EMG) demonstrated positive sharp waves and fibrillations in the left pelvic limb; the M wave of the left sciatic nerve was not obtainable by nerve stimulation. Seizures and dementia began during recovery from anesthesia. Six days after onset of paralysis, the dog was euthanized. Direct fluorescent antibody testing of the brain was positive for raccoon rabies virus. This case demonstrates clinical evaluation, CSF analysis, and EMG in an animal with rabies meningoencephalomyelitis. PMID- 14736721 TI - Serosurvey of antiBabesia antibodies in stray dogs and American pit bull terriers and American staffordshire terriers from North Carolina. AB - Stray dogs (n=359) and kennel dogs (n=149) from North Carolina were tested for evidence of antiBabesia antibodies. AntiBabesia antibodies were detected in 21/359 and 22/149 of the stray and kennel dogs, respectively. A total of 57 dogs from both groups were tested for babesiasis by light microscopy and polymerase chain reaction (PCR). Babesia deoxyribonucleic acid (DNA) was detected in 3/28 of the stray dogs and 14/29 of the kennel dogs. When Babesia DNA was detected by PCR, the species-specific PCR results differed from the Babesia species antibody titer results in 6/17 of the PCR-positive dogs. There was no association between antiBabesia antibodies and the presence of ticks. There are currently Babesia gibsoni epizootics affecting American pit bull terrier kennels. PMID- 14736722 TI - Clindamycin in the treatment of Babesia gibsoni infections in dogs. AB - This report examines the effectiveness of clindamycin for the treatment of babesiosis in dogs (n=10) experimentally infected with Babesia gibsoni (B. gibsoni). Clindamycin (25 mg/kg body weight, per os, q 12 hours for 14 days) gradually reduced parasitemia levels and induced morphological changes that indicated degeneration of parasites (e.g., segmentation; size reduction; localization in the cell limbic and/or torn state of the nucleus; and swelling, decrease, or disappearance of the cytoplasm) in the majority of dogs. Clindamycin treatment reduced the clinical symptoms characteristic of Babesia infection, including anemia, anorexia, and listlessness. Clindamycin might be useful as a medicine for treatment of B. gibsoni infection. PMID- 14736723 TI - Hematogenous septic arthritis in the dog: results of five patients treated nonsurgically with antibiotics. AB - This retrospective study evaluates the effectiveness of nonsurgical treatment using antibiotics to treat hematogenous septic arthritis in five dogs. Giant breed dogs were over-represented, with all dogs <1 year of age. Synovial fluid cultures were positive in all cases, with common bacterial species isolated that included Streptococcus B-haemolytic spp., Pasteurella multocida, and Staphylococcus intermedius. Dogs treated with appropriate duration and selection of antibiotics had clinical resolution with no residual deficits. This report and a previous clinical report demonstrate that hematogenous septic arthritis can be successfully treated nonsurgically with antibiotic therapy. PMID- 14736724 TI - External hydrocephalus in two cats. AB - External hydrocephalus describes an accumulation of cerebrospinal fluid (CSF) between the cerebral hemispheres and the overlying arachnoid membrane, rather than within the lateral ventricles. Two young cats with encephalopathic signs were diagnosed with external hydrocephalus, one via magnetic resonance imaging and one via computed tomography. Both cats had abnormally large, broad heads, with no evidence of open fontanelles. A surgical shunt was placed in each cat to divert the accumulated CSF within the cranial cavity to the peritoneal space. Both cats improved dramatically soon after surgical shunting was performed, and they continue to do well clinically, approximately 42 months and 8 months postoperatively, respectively. PMID- 14736725 TI - Results of single-session bilateral tibial plateau leveling osteotomies as a treatment for bilaterally ruptured cranial cruciate ligaments in dogs: 25 cases (2000-2001). AB - The records of 25 dogs with bilaterally ruptured cranial cruciate ligaments (CrCL) treated with single-session bilateral tibial plateau leveling osteotomies (TPLOs) were reviewed. The mean age and body weight at presentation were 4.9 years and 36.1 kg, respectively. Minor complications occurred in five dogs. Good or excellent function occurred in 19 of 25 dogs at a mean of 6.9 weeks postoperatively. Owner evaluation revealed that the 88% of dogs had good or excellent function at an average of 14.3 months postoperatively. This study demonstrated that single-session bilateral TPLOs are an effective treatment for bilaterally ruptured CrCLs with minimal morbidity and complications. PMID- 14736726 TI - Detailed structural features of glycan chains derived from alpha1-acid glycoproteins of several different animals: the presence of hypersialylated, O acetylated sialic acids but not disialyl residues. AB - We analyzed carbohydrate chains of human, bovine, sheep, and rat alpha1-acid glycoprotein (AGP) and found that carbohydrate chains of AGP of different animals showed quite distinct variations. Human AGP is a highly negatively charged acidic glycoprotein (pKa = 2.6; isoelectic point = 2.7) with a molecular weight of approximately 37,000 when examined by matrix-assisted laser-desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and contains di-, tri-, and tetraantennary carbohydrate chains. Some of the tri- and tetraantennary carbohydrate chains are substituted with a fucose residue (sialyl Lewis x type structure). In sheep AGP, mono- and disialo-diantennary carbohydrate chains were abundant. Tri- and tetrasialo-triantennary carbohydrate chains were also present as minor oligosaccharides, and some of the sialic acid residues were substituted with N-glycolylneuraminic acid. In rat AGP, very complex mixtures of disialo carbohydrate chains were observed. Complexity of the disialo-oligosaccharides was due to the presence of N, O-acetylneuraminic acids. Triantennary carbohydrate chains carrying N,O-acetylneuraminic acid were also observed as minor component oligosaccharides. We found some novel carbohydrate chains containing both N acetylneuraminic acid and N-glycolylneuraminic acid in bovine AGP. Interestingly, triantennary carbohydrate chains were hardly detected in bovine AGP, but diantennary carbohydrate chains with tri- or tetrasialyl residues were abundant. Furthermore the major sialic acid in these carbohydrate chains was N glycolylneuraminic acid. It should be noted that these sialic acids are attached to multiple sites of the core oligosaccharide and are not present as disialyl groups. PMID- 14736727 TI - Identification of carbohydrates binding to lectins by using surface plasmon resonance in combination with HPLC profiling. AB - A new, powerful method is presented for screening the binding in real time and taking place under dynamic conditions of oligosaccharides to lectins. The approach combines an SPR biosensor and HPLC profiling with fluorescence detection, and is applicable to complex mixtures of oligosaccharides in terms of ligand-fishing. Labeling the oligosaccharides with 2-aminobenzamide ensures a detection level in the fmol range. In an explorative study the binding of RNase B derived oligomannose-type N-glycans to biosensor-immobilized concanavalin A (Con A) was examined, and an affinity ranking could be established for Man(5)GlcNAc(2) to Man(9)GlcNAc(2), as monitored by HPLC. In subsequent experiments and using well-defined labeled as well as nonlabeled oligosaccharides, it was found that the fluorescent tag does not interfere with the binding and that the optimum epitope for the interaction with Con A comprises the tetramannoside unit Manalpha2Manalpha6(Manalpha3)Man[D(3)B(A)4'], rather than the generally accepted trimannoside Manalpha6 (Manalpha3)Man [B(A)4' or 4(4')3]. In a similar experimental setup, the interaction of various fucosylated human milk oligosaccharides with the fucose-binding lectin from Lotus tetragonolobus purpureaus was studied, and it appeared that oligosaccharides containing blood group H could selectively be retained and eluted from the lectin-coated surface. Finally, using the same lectin and a mixture of O-glycans derived from bovine submaxillary gland mucin, minor constituents but containing fucose could selectively be picked from the analyte solution as demonstrated by HPLC profiling. PMID- 14736728 TI - Mutation of the glycosylated asparagine residue 286 in human CLN2 protein results in loss of enzymatic activity. AB - Late infantile neuronal ceroid lipofuscinosis (LINCL) is caused by the deficiency of the lysosomal tripeptidyl peptidase-I encoded by CLN2. We previously detected in two LINCL patients a homozygous missense mutation, p.Asn286Ser, that affects a potential N-glycosylation site. We introduced the p.Asn286Ser mutation into the wild-type CLN2 cDNA and performed transient expression analysis to determine the effect on the catalytic activity, intracellular targeting, and glycosylation of the CLN2 protein. Expression of mutant p.Asn286Ser CLN2 in HEK293 cells revealed that the mutant was enzymatically inactive. Western blot analysis demonstrated that at steady state the amounts of expressed p.Asn286Ser CLN2 were reduced compared with wild-type expressing cells. The rate of synthesis and the sorting of the newly synthesized p.Asn286Ser CLN2 in the Golgi was not affected compared with wild-type CLN2 protein. The electrophoretic mobility of the immunoprecipitated mutant p.Asn286Ser CLN2 was increased by approximately 2 kDa compared with the wild-type CLN2 protein, whereas deglycosylation led to the generation of polypeptides of the same apparent size. The data suggest that mutant p.Asn286Ser CLN2 lacks one oligosaccharide chain resulting in enzymatic inactivation. PMID- 14736729 TI - Functional characterization of GumK, a membrane-associated beta glucuronosyltransferase from Xanthomonas campestris required for xanthan polysaccharide synthesis. AB - Xanthomonas campestris is a Gram-negative bacterium that produces an exopolysaccharide known as xanthan gum. Xanthan is involved in a variety of biological functions, including pathogenesis, and is widely used in the industry as thickener and viscosifier. Although the genetics and biosynthetic process of xanthan are well documented, the enzymatic components have not been examined and no data on glycosyltransferases have been reported. We describe the functional characterization of the gumK gene product, an essential protein for xanthan synthesis. Immunoblots and complementation studies showed that GumK is a 44-kDa protein associated to the membrane fraction. This value corresponds to the expected molecular mass for GumK encoded by an extended open reading frame than proposed from previous genetic data and in X. campestris published complete genome. The protein was expressed in Escherichia coli cells. The purified protein catalyzed the transfer of a glucuronic acid residue from UDP-glucuronic acid to mannose-alpha-1,3-glucose-beta-1,4-glucose-P-P-polyisoprenyl with formation of a glucuronic acid-beta-mannose linkage. We examined the acceptor substrate specificity. GumK was unable to use the trisaccharide acceptor freed from the pyrophosphate lipid moiety. Replacement of the natural lipid moiety by phytanyl showed that the catalytic function could proceed with glucuronic acid transfer. These results suggest the enzyme does not show specificity for the lipidic portion of the acceptor. GumK showed diminished activity when tested with 6-O acetyl-mannose-alpha-1,3-glucose-beta-1,4-glucose-P-P-polyisoprenyl, a putative intermediate in the synthesis of xanthan. This could indicate that acetylation of the internal mannose takes place after the formation of the GumK product. PMID- 14736730 TI - Tumor growth inhibition by indomethacin in a mouse model of human medullary thyroid cancer: implication of cyclooxygenases and 15-hydroxyprostaglandin dehydrogenase. AB - Medullary thyroid cancer (MTC) is a C cell neoplasm-secreting calcitonin. Surgery remains the only treatment as the primary tumor and metastases resist radio- and chemotherapies. MTC produces high amounts of prostaglandins (PGs). Nonsteroidal antiinflammatory drugs have an antitumoral effect, generally related to the decrease of PG levels. We assessed the therapeutic potential of indomethacin in a model of human (TT cells) tumors in nude mice. Indomethacin (1.5 or 2.0 mg/kg body weight.d for 7 wk) inhibited tumor volume by 49 or 77%, respectively, and decreased the plasma level of CT. Although the terminal deoxynucleotidyltransferase-mediated deoxyuridine triphosphate nick end labeling method revealed few apoptotic nuclei, the number of proliferating cells was significantly decreased (Ki-67 antigen study). Immunological effector recruitment and vascular network was not modified by treatment. The inducible synthesis enzyme, cyclooxygenase-2 (COX-2), was revealed only in infiltrating cells, both in treated and control tumors. The expression of the constitutive synthesis enzyme COX-1 was diminished, and the expression of 15-prostaglandin dehydrogenase, the key enzyme catabolizing PGs, was increased in treated tumors. Thus, our results demonstrated the potential of indomethacin, inhibitor of COX-1 and COX-2, to prevent MTC growth. The synthesis enzyme, COX-1, and the catabolism enzyme 15-prostaglandin dehydrogenase, could be involved in MTC development. PMID- 14736731 TI - Characterization of the rhesus monkey ghrelin gene and factors influencing ghrelin gene expression and fasting plasma levels. AB - Ghrelin stimulates release of GH from the pituitary, stimulates appetite, and may influence metabolic processes in other tissues expressing the GH secretagogue receptor. Ghrelin can thus influence behaviors and endocrine pathways contributing to weight gain. In this study we characterized the ghrelin gene from the rhesus monkey and analyzed the association of plasma ghrelin levels with metabolic and endocrine markers. Rhesus ghrelin is 97, 91, and 96% homologous to the human cDNA, gene, and peptide, respectively. Ghrelin expression was highest in the stomach with lower levels found in muscle and duodenum. In these tissues, ghrelin expression in calorie-restricted and obese animals was about 40-99% lower than in lean animals. In addition, ghrelin expression in muscle was fairly high and may allow this tissue to contribute significantly to plasma levels. Fasting plasma ghrelin concentrations were also inversely correlated with body mass index and exhibited a nonlinear association with age with increased levels in younger and older monkeys and lower levels in middle-aged monkeys. Although a significant inverse correlation between fasting plasma ghrelin and fasting insulin levels were found, iv glucose and insulin administration did not significantly alter ghrelin levels. These studies demonstrate that ghrelin levels are influenced by age-related factors and adiposity in the rhesus monkey. These similarities between the rhesus monkey and human ghrelin genes and plasma ghrelin responses suggest a unique opportunity to study the mechanisms regulating ghrelin secretion and gene expression in different tissues in normal and disease states using this model system. PMID- 14736732 TI - High expression of the R-type voltage-gated Ca2+ channel and its involvement in Ca2+-dependent gonadotropin-releasing hormone release in GT1-7 cells. AB - The GT1 cell has been widely used as a model cell to study cellular functions of GnRH neurons. Despite the importance of Ca(2+) channels, little is known except for L- and T-type Ca(2+) channels in GT1 cells. Therefore, we studied the diversity of voltage-gated Ca(2+) channels in GT1-7 cells with perforated-patch clamp and RT-PCR. An R-type Ca(2+) channel blocker, SNX-482, inhibited the Ca(2+) currents by 75.6% in all cells examined (n = 9). A T-type Ca(2+) channel blocker, Ni(2+), inhibited the Ca(2+) currents by 12.6% in all cells examined (n = 9). An L-type Ca(2+) channel blocker, nimodipine, inhibited the Ca(2+) currents by 17.9% in five of 11 cells examined. When using Ba(2+) as a charge carrier, another dihydropyridine antagonist, nifedipine, clearly inhibited the currents by 12.1% in all cells examined (n = 16). An N-type Ca(2+) channel blocker, omega-conotoxin GVIA, inhibited the Ca(2+) currents by 13.8% in three of 20 cells examined. A P/Q type Ca(2+) channel blocker, omega-agatoxin-IVA, had no effect on the currents (n = 9). RT-PCR revealed that GT1-7 cells expressed the alpha(1B), alpha(1D), alpha(1E), and alpha(1H) subunit mRNA. Furthermore, SNX-482 and nifedipine inhibited the high K(+)-induced increase in the intracellular Ca(2+) concentration and GnRH release. omega-Conotoxin-GVIA and omega-agatoxin-IVA had no effect. These results suggest that GT1-7 cells express R-, L-, N-, and T-type voltage-gated Ca(2+) channels; the R-type was a major current component, and the L-, N-, and T-types were minor ones. The R- and L-type Ca(2+) channels play a critical role in the regulation of Ca(2+)-dependent GnRH release. PMID- 14736733 TI - Ventricular nonmyocytes inhibit doxorubicin-induced myocyte apoptosis: involvement of endogenous endothelin-1 as a paracrine factor. AB - A cross-talk between cardiac myocytes and nonmyocytes via humoral factors plays an important role in the development of cardiac growth. However, it remains to be elucidated whether humoral factors produced from nonmyocytes have a protective effect on acute myocardial injury. The present in vitro study investigated the antiapoptotic effect of nonmyocytes on doxorubicin (DOX)-induced myocyte apoptosis and its molecular mechanism. Myocyte-nonmyocyte coculture and treatment with nonmyocyte-conditioned media significantly attenuated DOX-induced myocyte apoptosis. Treatment with nonmyocyte-conditioned media stimulated the phosphorylation of ERK, Akt, and cAMP response element-binding protein (CREB) in myocytes. Nonmyocyte-conditioned media also increased protein levels of Bcl-2 but not Bcl-xL and decreased caspase-3 activation induced by DOX. MAPK kinase specific inhibitor PD98059, phosphatidylinositol-3 kinase-Akt inhibitor LY294002, and CREB antisense oligonucleotide significantly blocked the antiapoptotic effect of nonmyocyte-conditioned media. A considerable amount of endothelin (ET)-1 production was detected in nonmyocytes but not in myocytes. Exogenous ET-1 mimicked nonmyocyte-conditioned media-mediated ERK and CREB phosphorylation and Bcl-2 protein increase but not Akt phosphorylation. In addition, ET-A receptor antagonists BQ123 and BQ485 partially blocked nonmyocyte-conditioned media mediated antiapoptotic effect, ERK and CREB phosphorylation, and Bcl-2 protein increase. Nonmyocyte-conditioned media and exogenous ET-1 unchanged protein levels of manganese superoxide dismutase and oxidative stress-related product levels augmented by DOX. The present findings demonstrate that cardiac nonmyocytes inhibit DOX-induced myocyte apoptosis, at least in part, via ET-1 secretion-mediated CREB activation independent of the decrease in oxidative stress. PMID- 14736734 TI - Differential expression of steroidogenic factor-1/adrenal 4 binding protein and liver receptor homolog-1 (LRH-1)/fetoprotein transcription factor in the rat testis: LRH-1 as a potential regulator of testicular aromatase expression. AB - Aromatase converts testicular androgens to estrogens, which are essential for male fertility. Aromatase expression in testis occurs via transcription from promoter II, and requires the presence of a nuclear receptor half-site that binds the orphan receptor steroidogenic factor-1 [SF-1 (nuclear receptor 5A1)] to mediate basal and (in part) cAMP-induced transcription. We hypothesized that liver receptor homolog-1 (LRH-1) (nuclear receptor 5A2), a receptor closely related to SF-1, could also play a role in regulating aromatase expression in the testis. We demonstrate expression of LRH-1 in adult rat and immature mouse Leydig cells (LHR-1 > SF-1) as well as in pachytene spermatocytes and round spermatids but not in Sertoli cells, which in contrast, express high levels of SF-1. In transient transfection assays using TM3 Leydig cells and TM4 Sertoli cells, a rat promoter II luciferase reporter construct was stimulated by cotransfection of LRH 1 expression vector. Mutation analysis showed that induction by LRH-1 in TM3 and TM4 cells requires an AGGTCA motif at position -90, to which LRH-1 bound in gel shift analysis. We therefore provide evidence that LRH-1 plays an important role in the regulation of aromatase expression in Leydig cells. The colocalization of LRH-1 and aromatase to multiple testis cell types suggests that LRH-1 may have important effects on estrogen production, testis development, spermatogenesis, and testicular carcinogenesis. PMID- 14736735 TI - Extracellular signal-regulated kinase, Jun N-terminal kinase, p38, and c-Src are involved in gonadotropin-releasing hormone-stimulated activity of the glycoprotein hormone follicle-stimulating hormone beta-subunit promoter. AB - The role of ERK, Jun N-terminal kinase (JNK), p38, and c-Src in GnRH-stimulated FSHbeta-subunit promoter activity was examined in the LbetaT-2 gonadotroph cell line. Incubation of the cells with a GnRH agonist resulted in activation of ERK, JNK, p38, and c-Src. The peak of ERK activation was observed at 5 min, whereas that of JNK, p38, and c-Src at 30 min, declining thereafter. ERK activation by GnRH is dependent on protein kinase C (PKC), as evident by activation, inhibition, and depletion of 12-O-tetradecanoylphorbol-13-acetate-sensitive PKC subspecies. Ca(2+) influx, but not Ca(2+) mobilization, is required for ERK activation. GnRH signaling to ERK is partially mediated by dynamin and a protein tyrosine kinase, apparently c-Src. ERK activation by GnRH in LbetaT-2 cells does not involve transactivation of epidermal growth factor receptor or mediation via Gbetagamma or beta-arrestin. Once activated by GnRH, ERK translocates to the nucleus. We examined the role of ERK, JNK, p38, and c-Src in GnRH-stimulated ovine FSHbeta promoter, linked to a luciferase reporter gene (-4741oFSHbeta-LUC). The PKC activator 12-O-tetradecanoylphorbol-13-acetate, but not the Ca(2+) ionophore ionomycin, stimulated FSHbeta-luciferase (LUC) activity. Furthermore, down-regulation of PKC, but not removal of Ca(2+), inhibited the GnRH response. Cotransfection of FSHbeta-LUC and the constitutively active forms of Raf-1 and MEK stimulated FSHbeta-LUC activity, whereas the dominant negatives of Ras, Raf 1, and MEK and the selective MEK inhibitor PD98059, abolished GnRH-induced FSHbeta-LUC activity. The dominant negatives of CDC42 and JNK reduced the GnRH response by 36 and 49%, respectively. Incubation of the cells with the p38 or the c-Src inhibitors SB203580 and PP1 also reduced the GnRH response. Surprisingly, two proximal activator protein-1 sites contribute very little to the GnRH response. Thus, PKC, ERK, JNK, p38, and c-Src, but not Ca(2+), are involved in GnRH induction of the ovine FSHbeta gene. PMID- 14736736 TI - Urocortin II gene is highly expressed in mouse skin and skeletal muscle tissues: localization, basal expression in corticotropin-releasing factor receptor (CRFR) 1- and CRFR2-null mice, and regulation by glucocorticoids. AB - Peptides encoded by the Urocortin (Ucn) II gene, also known as stresscopin related peptide, were recently identified as new members of the corticotropin releasing factor (CRF) family. Ucn II is a specific ligand for the type 2 CRF receptor (CRFR). We have demonstrated the peripheral distribution of mouse Ucn (mUcn) II transcripts by using specific mUcn II ribonuclease protection assays, RT-PCR, Southern hybridization, and DNA sequencing. Although Ucn II mRNA is widely expressed in a variety of peripheral tissues, we found it to be most highly expressed in the skin and skeletal muscle tissues. Using a specific RIA for mUcn II, we detected Ucn II-like immunoreactivity (ir) in acid extracts of mouse brain, muscle, and skin. Immunohistochemical studies revealed Ucn II-like ir in both skin epidermis and adnexal structures and in the skeletal muscle myocytes. Ucn II mRNA and ir were also observed in neonatal skeletal muscle cultures in which Ucn II was localized to the myotube. We found a significant increase in Ucn II mRNA levels in the skin, but not in skeletal muscle, of both CRFR1- and CRFR2-null mice compared with their wild-type littermates. We showed that administration of dexamethasone to mice resulted in a decrease of Ucn II mRNA levels in the back skin region 12 h after ip injections. Removal of the adrenal gland significantly increased the levels of Ucn II mRNA in the skin, and the levels were reduced back to normal levels after corticosterone replacement. Further examination of the distribution and regulation of CRFR2 and its specific ligand Ucn II in the skin and skeletal muscle tissues may reveal the manner by which the CRFR2 pathway is involved in the physiological responses to stress in these tissues and in other pathophysiologies of the skin and muscle. PMID- 14736737 TI - Prostaglandin biosynthesis, transport, and signaling in corpus luteum: a basis for autoregulation of luteal function. AB - The corpus luteum (CL) is a transient ovarian endocrine gland formed from the ovulated follicle. Progesterone is the primary secretory product of CL and is essential for establishment of pregnancy in mammals. In the cyclic female, the life span of CL is characterized by luteal development, maintenance, and regression regulated by complex interactions between luteotrophic and luteolytic mediators. It is universally accepted that prostaglandin (PG) F(2a) is the luteolysin whereas PGE(2) is considered as a luteotropin in most mammals. New emerging concepts emphasize the autocrine and paracrine actions of luteal PGs in CL function. However, there is no report on selective biosynthesis and cellular transport of luteal PGE(2) and PGF(2alpha) in the CL of any species. We have studied the expression of enzymes involved in the metabolism of PGE(2) and PGF(2alpha), cyclooxygenase (COX)-1 and -2, PGE and F synthases, PG 15 dehydrogenase, and PG transporter as well as receptors (EP2, EP3, and FP) throughout the CL life span using a bovine model. COX-1, PGF synthase, and PG 15 dehydrogenase are expressed at constant levels whereas COX-2, PGE synthase, PG transporter, EP2, EP3, and FP are highly modulated during different phases of the CL life span. The PG components are preferentially expressed in large luteal cells. The results indicate that PGE(2) biosynthesis, transport, and signaling cascades are selectively activated during luteal maintenance. By contrast the PGF(2alpha) system is activated during luteal regression. Collectively, our results suggest an integrated role for luteal PGE(2) and PGF(2alpha) in autoregulation of CL function. PMID- 14736738 TI - Stress-induced suppression of the gonadotropin-releasing hormone pulse generator in the female rat: a novel neural action for calcitonin gene-related peptide. AB - In addition to its role as a potent vasodilator, calcitonin gene-related peptide (CGRP) is centrally involved in a variety of stress responses, including activation of the hypothalamo-pituitary-adrenocortical axis. It is well known that stress suppresses the activity of the hypothalamic GnRH pulse generator, the central regulator of LH and FSH pulses, resulting in reproductive dysfunction. The aim of this study was to test the hypothesis that CGRP has a critical role in mediating stress-induced suppression of pulsatile LH secretion in the rat. Ovariectomized rats were implanted with intracerebroventricular and iv cannulae. Central administration of CGRP (75 pmol-1.2 nmol) into the lateral cerebral ventricle resulted in a profound, dose-dependent suppression of LH pulses, which was reversed by a CGRP receptor antagonist (CGRP(8-37),1 nmol). Although the site of action of CGRP remains to be established, the induction of c-Fos expression in the preoptic area and hypothalamic paraventricular nucleus might suggest an involvement of these brain regions. Intravenous administration of CGRP did not affect LH pulses. Coadministration (intracerebroventricular) of CGRP (400 pmol) with a CRH antagonist (alpha-helical CRF(9-41), 26 nmol) partly blocked the CGRP induced suppression of LH pulses. Furthermore, CGRP(8-37) (1 nmol) completely blocked hypoglycemic stress-induced suppression of LH pulses. These results suggest that the suppression of pulsatile LH secretion by central administration of CGRP may be mediated in part by CRH, and that CGRP may play a pivotal role in the normal physiological response of stress-induced suppression of the hypothalamic GnRH pulse generator, and hence the reproductive system. PMID- 14736739 TI - Glucocorticoids delay age-associated thymic involution through directly affecting the thymocytes. AB - After puberty, the thymus undergoes a dramatic loss in size, resulting in a reduction in the number of thymocytes, a phenomenon termed age-associated thymic involution. The factors regulating this process are poorly understood. We investigated the role of endogenous glucocorticoids (GCs) in this process by studying transgenic mice with increased GC sensitivity restricted to the T-cell lineage due to overexpression of a GC-receptor transgene under the control of the proximal lck promoter. Surprisingly, in these transgenic mice, the age-associated thymic involution did not start until after 6 months of age, demonstrating that endogenous GCs through directly affecting the thymocytes delay the age-associated thymic involution process. The delayed age-associated thymic involution resulted in a significantly higher number of thymocytes in transgenic mice, compared with wild-type mice at 6 months of age or older. The higher number of thymocytes was associated with increased percentage of cycling double-negative and single positive thymocytes, whereas thymic apoptosis was unaffected. The above effects of GCs were restricted to the thymocytes and were not reflected on the peripheral T cells, in which GCs suppressed the number of peripheral T cells in aged transgenic mice, demonstrating that thymocytes and T cells are differentially regulated by GCs. Furthermore, CD4(+) T cells were more affected than CD8(+) T cells, resulting in a decrease in the CD4/CD8 T-cell ratio. In summary, our results reveal novel biological effects of endogenous GCs on thymic involution and T-cell homeostasis in aged mice. PMID- 14736740 TI - A role for thyroid hormone in wound healing through keratin gene expression. AB - The importance of thyroid hormone (TH) in wound healing is not well understood. To gain insight, we evaluated the impact of TH deficiency on wound-healing genes in cultured keratinocytes. By RT-PCR, keratin 6a (K6a) and 16 (K16) gene expression in TH replete cells was 3.8- (P < 0.005) and 1.9-fold (P < 0.05) greater, respectively, than expression in TH-deficient cells. By real-time PCR, TH replete cell expression of K6a, K16, and K17 was greater than in deficient cells: 18- (P < 0.001), 10- (P < 0.001), and 4-fold (P < 0.005), respectively. To examine TH requirement for optimal wound healing, we contrasted TH-deficient vs. ip T(3)-treated mice. Four days after wounding, ip T(3)-treated mice had twice the degree of wound closure as hypothyroid mice (P < 0.001). By RT-PCR, K6a and K17 gene expression from control mouse skin was greater than from hypothyroid mouse skin: 5- (P < 0.001) and 1.7-fold (P < 0.05), respectively. T(3) is necessary for the keratinocyte proliferation required for optimal wound healing. T(3) exerts influence by stimulating expression of the wound-healing keratin genes. Thus, for hypothyroid patients undergoing surgery that cannot be delayed until euthyroidism is achieved, our data support T(3) treatment for the perioperative period. PMID- 14736741 TI - A central role for the notochord in vertebral patterning. AB - The vertebrates are defined by their segmented vertebral column, and vertebral periodicity is thought to originate from embryonic segments, the somites. According to the widely accepted 'resegmentation' model, a single vertebra forms from the recombination of the anterior and posterior halves of two adjacent sclerotomes on both sides of the embryo. Although there is supporting evidence for this model in amniotes, it remains uncertain whether it applies to all vertebrates. To explore this, we have investigated vertebral patterning in the zebrafish. Surprisingly, we find that vertebral bodies (centra) arise by secretion of bone matrix from the notochord rather than somites; centra do not form via a cartilage intermediate stage, nor do they contain osteoblasts. Moreover, isolated, cultured notochords secrete bone matrix in vitro, and ablation of notochord cells at segmentally reiterated positions in vivo prevents the formation of centra. Analysis of fss mutant embryos, in which sclerotome segmentation is disrupted, shows that whereas neural arch segmentation is also disrupted, centrum development proceeds normally. These findings suggest that the notochord plays a key, perhaps ancient, role in the segmental patterning of vertebrae. PMID- 14736742 TI - Endothelial cell interactions initiate dorsal pancreas development by selectively inducing the transcription factor Ptf1a. AB - Dorsal and ventral pancreatic bud development from the endoderm requires inductive interactions with diverse mesodermal cell types and the action of transcription factors expressed within the endoderm. Presently it is unclear which mesodermal interactions activate which pancreatic transcription factors, and whether such inductions are common for initiating dorsal and ventral pancreas development. Previous studies of Lammert et al. showed that signaling from embryonic blood vessel cells, derived from the mesoderm, promotes pancreatic bud development. Using a combination of mouse Flk1(-/-) embryos lacking endothelial cells and tissue recombination experiments, we discovered that the initial induction of dorsal endoderm cells positive for the pancreatic and duodenal transcription factor Pdx1 does not require aorta or endothelial cell interactions, but dorsal pancreatic bud emergence and the maintenance of Pdx1 expression does. Aortal endothelial cells induce the crucial pancreatic transcription factor Ptf1a in the dorsal pancreatic endoderm; whereas the vitelline veins, which are normally adjacent to the emerging ventral pancreatic bud, are unnecessary for ventral Ptf1a induction or for ventral pancreatic bud initiation. We find that the aorta cells themselves, apart from the blood supply, cause the induction of Ptf1a in dorsal endoderm explants. Thus, endothelial cell interactions specifically promote early dorsal pancreatic development, at least in part, by inducing Ptf1a(+) pancreatic progenitors. Additionally, we find that endothelial cells are necessary for the induction of both the insulin and glucagon genes. PMID- 14736743 TI - The deafness gene dfna5 is crucial for ugdh expression and HA production in the developing ear in zebrafish. AB - Over 30 genes responsible for human hereditary hearing loss have been identified during the last 10 years. The proteins encoded by these genes play roles in a diverse set of cellular functions ranging from transcriptional regulation to K(+) recycling. In a few cases, the genes are novel and do not give much insight into the cellular or molecular cause for the hearing loss. Among these poorly understood deafness genes is DFNA5. How the truncation of the encoded protein DFNA5 leads to an autosomal dominant form of hearing loss is not clear. In order to understand the biological role of Dfna5, we took a reversegenetic approach in zebrafish. Here we show that morpholino antisense nucleotide knock-down of dfna5 function in zebrafish leads to disorganization of the developing semicircular canals and reduction of pharyngeal cartilage. This phenotype closely resembles previously isolated zebrafish craniofacial mutants including the mutant jekyll. jekyll encodes Ugdh [uridine 5'-diphosphate (UDP)-glucose dehydrogenase], an enzyme that is crucial for production of the extracellular matrix component hyaluronic acid (HA). In dfna5 morphants, expression of ugdh is absent in the developing ear and pharyngeal arches, and HA levels are strongly reduced in the outgrowing protrusions of the developing semicircular canals. Previous studies suggest that HA is essential for differentiating cartilage and directed outgrowth of the epithelial protrusions in the developing ear. We hypothesize that the reduction of HA production leads to uncoordinated outgrowth of the canal columns and impaired facial cartilage differentiation. PMID- 14736744 TI - Hex homeobox gene-dependent tissue positioning is required for organogenesis of the ventral pancreas. AB - In animal development, digestive tissues emerge from different positions of the endoderm as a result of patterning signals from overlying mesoderm. Although embryonic tissue movement during gastrulation generates an initial positional relationship between the endoderm and mesoderm, the role of subsequent endoderm movement against the mesoderm in patterning is unknown. At embryonic day 8.5 in the mouse, proliferation of cells at the leading edge of ventral-lateral endoderm, where the liver and ventral pancreas emerge, helps close off the foregut. During this time, the endoderm grows adjacent to and beyond the cardiogenic mesoderm, an inducer of the liver program and an inhibitor of the pancreas program. The homeobox gene Hex is expressed in this endoderm cell domain and in the liver and ventral pancreas buds, after organogenesis. We have found that in Hex(-/-) embryos, there is a complete failure in ventral pancreatic specification, while the liver program is still induced. However, when Hex-null ventral endoderm is isolated prior to its interaction with cardiogenic mesoderm and is cultured in vitro, it activates early pancreas genes. We found that Hex controls the proliferation rate, and thus the positioning, of the leading edge of endoderm cells that grow beyond the cardiogenic mesoderm, during gut tube closure. Thus, Hex-controlled positioning of endoderm cells beyond cardiogenic mesoderm dictates ventral pancreas specification. Other endodermal transcription factors may also function morphogenetically rather than by directly regulating tissue-specific programs. PMID- 14736745 TI - The murine winged-helix transcription factor Foxl2 is required for granulosa cell differentiation and ovary maintenance. AB - Human Blepharophimosis/ptosis/epicanthus inversus syndrome (BPES) type I is an autosomal dominant disorder associated with premature ovarian failure (POF) caused by mutations in FOXL2, a winged-helix/forkhead domain transcription factor. Although it has been shown that FOXL2 is expressed in adult ovaries, its function during folliculogenesis is not known. Here, we show that the murine Foxl2 gene is essential for granulosa cell differentiation and ovary maintenance. In Foxl2(lacZ) homozygous mutant ovaries granulosa cells do not complete the squamous to cuboidal transition leading to the absence of secondary follicles and oocyte atresia. We further demonstrate that activin-betaA and anti-Mullerian inhibiting hormone expression is absent or strongly diminished in Foxl2(lacZ) homozygous mutant ovaries. Unexpectedly, two weeks after birth most if not all oocytes expressed Gdf9 in Foxl2(lacZ) homozygous mutant ovaries, indicating that nearly all primordial follicles have already initiated folliculogenesis at this stage. This activation, in the absence of functional granulosa cells, leads to oocyte atresia and progressive follicular depletion. In addition to providing a molecular mechanism for premature ovarian failure in BPES, these results suggest that granulosa cell function is not only crucial for oocyte growth but also to maintain follicular quiescence in vivo. PMID- 14736746 TI - Mili, a mammalian member of piwi family gene, is essential for spermatogenesis. AB - The piwi family genes, which are defined by conserved PAZ and Piwi domains, play important roles in stem cell self-renewal, RNA silencing, and translational regulation in various organisms. To reveal the function of the mammalian homolog of piwi, we produced and analyzed mice with targeted mutations in the Mili gene, which is one of three mouse homologs of piwi. Spermatogenesis in the MILI-null mice was blocked completely at the early prophase of the first meiosis, from the zygotene to early pachytene, and the mice were sterile. However, primordial germ cell development and female germ cell production were not disturbed. Furthermore, MILI bound to MVH, which is an essential factor during the early spermatocyte stage. The similarities in the phenotypes of the MILI- and MVH-deficient mice and in the physical binding properties of MILI and MVH indicate a functional association of these proteins in post-transcriptional regulation. These data indicate that MILI is essential for the differentiation of spermatocytes. PMID- 14736747 TI - Identification of minimal enhancer elements sufficient for Pax3 expression in neural crest and implication of Tead2 as a regulator of Pax3. AB - Pax3 is a transcription factor that is required by pre-migratory neural crest cells, which give rise to the peripheral nervous system, melanocytes, some vascular smooth muscle, and numerous other derivatives [corrected]. Both mice and humans with Pax3 deficiency exhibit neural crest-related developmental defects [corrected]. Pax3 is also expressed in the dorsal neural tube, and by myogenic progenitors in the presomitic mesoderm and the hypaxial somites. Molecular pathways that regulate Pax3 expression in the roof plate probably represent early upstream signals in neural crest induction. We have identified an enhancer region in the Pax3 genomic locus that is sufficient to recapitulate expression in neural crest precursors in transgenic mice. We show that Tead2, a member of the Tead box family of transcription factors, binds to a neural crest enhancer and activates Pax3 expression. Tead2, and its co-activator YAP65, are co-expressed with Pax3 in the dorsal neural tube, and mutation of the Tead2 binding site in the context of Pax3 transgenic constructs abolishes neural expression. In addition, a Tead2 Engrailed fusion protein is able to repress retinoic acid-induced Pax3 expression in P19 cells and in vivo. These results suggest that Tead2 is an endogenous activator of Pax3 in neural crest. PMID- 14736748 TI - XSEB4R, a novel RNA-binding protein involved in retinal cell differentiation downstream of bHLH proneural genes. AB - RNA-binding proteins play key roles in the post-transcriptional regulation of gene expression but so far they have not been studied extensively in the context of developmental processes. We report on the molecular cloning and spatio temporal expression of a novel RNA-binding protein, XSEB4R, which is strongly expressed in the nervous system. This study is focused on the analysis of Xseb4R in the context of primary neurogenesis and retinogenesis. To study Xseb4R function during eye development, we set up a new protocol allowing in vivo lipofection of antisense morpholino oligonucleotides into the retina. The resulting XSEB4R knockdown causes an impairment of neuronal differentiation, with an increase in the number of glial cells. By contrast, our gain-of-function analysis demonstrates that Xseb4R strongly promotes neural differentiation. We also showed a similar function during primary neurogenesis. Consistent with this proneural effect, we found that in the open neural plate Xseb4R expression is upregulated by the proneural gene XNgnr1, as well as by the differentiation gene XNeuroD, but is inhibited by the Notch/Delta pathway. Altogether, our results suggest for the first time a proneural effect for a RNA-binding protein involved in the genetic network of retinogenesis. PMID- 14736749 TI - Misrouting of mitral cell progenitors in the Pax6/small eye rat telencephalon. AB - The olfactory bulb is a protruding structure formed at the rostral end of the telencephalon. Pax6-mutant mice and rats lack the olfactory bulb and, instead, develop an olfactory bulb-like structure at the lateral part of the telencephalon. Here, we report that ectopic formation of the olfactory bulb-like structure in these mutants is caused by the abnormal migration of mitral cell progenitors, which first differentiate within the olfactory bulb. Cell-tracing experiments in whole embryos in culture indicate that, in the mutants, the mitral cell progenitors that originate from the rostral part of the telencephalon migrate caudally toward the lateral part of the telencephalon. Cell transplantation demonstrates that the abnormal cell migration is not autonomous to the mitral cell progenitors themselves. The mislocation of the olfactory bulb in the mutant is not caused by loss of olfactory nerve innervation. Furthermore, transfection of a Pax6-expression vector to the mutant telencephalon restores the normal migration of mitral cell progenitors. These results provide evidence that Pax6 is required to position the mitral cell progenitors at the rostral end of the telencephalon. PMID- 14736750 TI - src64 and tec29 are required for microfilament contraction during Drosophila cellularization. AB - Formation of the Drosophila cellular blastoderm involves both membrane invagination and cytoskeletal regulation. Mutations in src64 and tec29 reveal a novel role for these genes in controlling contraction of the actin-myosin microfilament ring during this process. Although membrane invagination still proceeds in mutant embryos, its depth is not uniform, and basal closure of the cells does not occur during late cellularization. Double-mutant analysis between scraps, a mutation in anillin that eliminates microfilament rings, and bottleneck suggests that microfilaments can still contract even though they are not organized into rings. However, the failure of rings to contract in the src64 bottleneck double mutant suggests that src64 is required for microfilament ring contraction even in the absence of Bottleneck protein. Our results suggest that src64-dependent microfilament ring contraction is resisted by Bottleneck to create tension and coordinate membrane invagination during early cellularization. The absence of Bottleneck during late cellularization allows src64-dependent microfilament ring constriction to drive basal closure. PMID- 14736751 TI - Eye movement and visuomotor arm movement deficits following mild closed head injury. AB - Based on increasing evidence that even mild closed head injury (CHI) can cause considerable neural damage throughout the brain, we hypothesized that mild CHI will disrupt the complex cerebral networks concerned with oculomotor and upper limb visuomotor control, resulting in impaired motor function. Within 10 days following mild CHI (Glasgow Coma Scale 13-15, alteration of consciousness <20 min), we compared 30 patients (15-37 years) and 30 matched controls on different types of saccades, oculomotor smooth pursuit (sine and random), upper-limb visuomotor performance and several neuropsychological tests known to be sensitive to head trauma. Simple reflexive saccades were not impaired, whereas, on the antisaccade task, the CHI group demonstrated prolonged saccadic latencies, a marginally higher number of directional errors and poorer spatial accuracy. The CHI group exhibited more directional errors and impaired motor accuracy on memory guided sequences of saccades and produced fewer self-paced saccades within 30 s. Most measures of sinusoidal and random oculomotor smooth pursuit showed no deficits, with the exception of a prolonged lag on random smooth pursuit in the CHI group. While arm movement reaction time and arm steadiness were not impaired, the CHI group showed decreased arm movement speed and decreased upper-limb motor accuracy. Conversely, after controlling for IQ, the CHI group had few head trauma related neuropsychological deficits. These results indicate that multiple motor systems can be impaired following mild CHI and that this can occur independently of neuropsychological impairment. Our study also indicates that quantitative tests of oculomotor and upper-limb visuomotor function may provide sensitive markers of cerebral dysfunction, suggesting the potential use of such tests to supplement patient assessment. To our knowledge, this study is the first to demonstrate the presence of oculomotor or visuomotor deficits following mild CHI. PMID- 14736752 TI - Differential impact of the FMR1 gene on visual processing in fragile X syndrome. AB - Fragile X syndrome (FXS) is the most common form of heritable mental retardation, affecting approximately 1 in 4000 males. The syndrome arises from expansion of a trinucleotide repeat in the 5'-untranslated region of the fragile X mental retardation 1 (FMR1) gene, leading to methylation of the promoter sequence and lack of the fragile X mental retardation protein (FMRP). Affected individuals display a unique neurobehavioural phenotype that includes striking visual-motor deficits. Here we provide neurobiological and behavioural evidence that supports the hypothesis that these visual-motor deficits are attributable to a magnocellular (M) visual pathway impairment. Immunohistochemical staining of a lateral geniculate nucleus (LGN) of a normal human male revealed high FMRP basal expression selectively within the M layers, suggesting an increased susceptibility of these neurons to the lack of FMRP as occurs in FXS. Similar staining of monkey LGNs for quantification purposes revealed that the difference is not an artefact of cell size differences between M and parvocellular (P) neurons. Further, Nissl staining of the LGNs of a male FXS patient revealed alaminar nuclei comprised of a homogenous population of small sized neurons, providing anatomical and morphological support for the idea that an M pathway pathology exists in FXS. Consistent with these neurobiological data, we have found that male patients with FXS have reduced sensitivity for psychophysical stimuli that probe the M pathway but not for those that probe the P pathway, a complementary visual stream that performs a separate set of early visual operations. Finally, male patients with FXS performed poorly on a global motion task but not on a form perception task, suggesting that the M pathway thalamic deficit may have a selective impact on cortical visual functioning in the parietal lobe, which is known to be a major recipient of M pathway afferents via the primary visual cortex. Together, these findings provide the first evidence that the loss of a single gene product, FMRP, in humans leads to abnormal neuroanatomical morphology of the LGN and a concomitant selective visual deficit of the M pathway. PMID- 14736753 TI - Subthalamic nucleus stimulation and dysarthria in Parkinson's disease: a PET study. AB - In Parkinson's disease, functional imaging studies during limb motor tasks reveal cerebral activation abnormalities that can be reversed by subthalamic nucleus (STN) stimulation. The effect of STN stimulation on parkinsonian dysarthria has not, however, been investigated using PET. The aim of the present study was to evaluate the effect of STN stimulation on regional cerebral blood flow (rCBF) during speech production and silent articulation in patients with Parkinson's disease. Ten Parkinson's disease patients surgically implanted bilaterally in the STN and with significant improvement of their dysarthria induced by STN stimulation were included. Ten healthy control subjects also participated in this study. Control subjects performed six sessions of [15O]H2O-PET scanning corresponding to three duplicated conditions externally cued by an auditory signal. The conditions were: (i) rest; (ii) production of a short, simple sentence; and (iii) silent articulation of the same sentence. Parkinson's disease patients carried out the six PET sessions twice, i.e., in the ON and OFF STN stimulation states. PET data analysis was performed using statistical parametric mapping (SPM99). In control subjects, speech production (SP) compared with rest was associated with increased rCBF bilaterally in the primary motor cortex (M1) corresponding to the orofacial somatotopy, the supplementary motor area (SMA), the associative auditory cortex and the cerebellar hemispheres. Silent articulation (SA) compared with rest induced a bilateral rCBF increase restricted to the orofacial M1 and cerebellar hemispheres. In Parkinson's disease patients in the OFF stimulation condition, during both SP and SA there was a lack of activation in the right orofacial M1 and in the cerebellum, abnormal increased rCBF in the right superior premotor cortex, and overactivation of the SMA. There was also an abnormal, increased rCBF in the dorsolateral prefrontal cortex (DLPFC) only during SP and increased rCBF in the left insula only during SA. In Parkinson's disease patients ON stimulation, for both SP and SA the activation pattern appeared similar to that in control subjects. In conclusion, our results suggest that parkinsonian dysarthria is associated with altered recruitment of the main motor cerebral regions (orofacial M1, cerebellum), and increased involvement of the premotor and prefrontal cortices (DLPFC, SMA, superior premotor cortex). These abnormal activations are different from those reported during hand motor tasks. They could be a compensatory mechanism, but might also arise directly as part of the pathophysiology of Parkinson's disease. STN stimulation tends to reverse these abnormal activations, which is consistent with the observed improvement of Parkinson's disease dysarthria. PMID- 14736754 TI - Serial magnetization transfer imaging in acute optic neuritis. AB - In serial studies of multiple sclerosis lesions, reductions in magnetization transfer ratio (MTR) are thought to be due to demyelination and axonal loss, with later rises due to remyelination. This study followed serial changes in MTR in acute optic neuritis in combination with clinical and electrophysiological measurements to determine if the MTR changes over time mirror the picture in multiple sclerosis lesions, further validating MTR as a marker of tissue integrity. Twenty-nine patients were recruited who had acute optic neuritis for a median of 13 days (range 7-24 days) since the onset of visual symptoms. A clinical examination and measurement of visual evoked potentials (VEP) was performed on each patient. Their optic nerves were imaged with a fat-saturated fast spin echo (FSE) sequence and a magnetization transfer sequence. Twenty-one had multiple subsequent examinations over the course of 1 year. In addition, 27 control subjects had their optic nerves imaged up to three times over 1 year. A blinded observer segmented the optic nerves from the MTR maps. Lesions were defined on the acute FSE images and, from the coordinates, the ratio of mean lesion MTR : healthy nerve MTR (lesion ratio) was calculated for each dataset. The time-averaged mean MTR in control optic nerves was 47.7 per cent units (pu). In diseased optic nerves, baseline mean MTR was 47.3 pu, with a mean lesion ratio of 0.98. The diseased optic nerve MTR and lesion ratio declined over time with a nadir at about 240 days at a mean MTR value of 44.2 pu and mean lesion ratio of 0.91. Subsequently, diseased optic nerve MTR appeared to rise; after 1 year the diseased optic nerve mean MTR was 45.1 pu (mean lesion ratio 0.93), although the difference was not significant compared with the nadir value. For each 0.01 increase in time-averaged lesion ratio logMAR visual acuity recovery improved by 0.03 (95% CI, 0.002, 0.08, P = 0.02). Time-averaged VEP central field latency was shorter by 6.1 ms (95% CI 1.5, 10.7, P = 0.012) per 1 pu rise in time-averaged diseased optic nerve MTR. The early fall in diseased optic nerve MTR is consistent with demyelination and Wallerian degeneration of transected axons. The late nadir compared with studies of multiple sclerosis lesions may have been due to slow clearance of myelin debris. Remyelination may have influenced subsequent MTR changes. The observations support using MTR to monitor symptomatic demyelinating lesions. PMID- 14736755 TI - Frequency and phenotypic spectrum of ataxia with oculomotor apraxia 2: a clinical and genetic study in 18 patients. AB - Ataxia with oculomotor apraxia type 2 (AOA2) is a newly described autosomal recessive cerebellar ataxia (ARCA) defined by genetic location to 9q34 of three families sharing gait ataxia, oculomotor apraxia and/or elevated alpha foetoprotein (AFP) levels. We have evaluated 77 families with progressive non Friedreich ARCA and have identified six families with a phenotype suggestive of AOA2. Linkage was confirmed in all six families, with a maximal lod score of 5.91 at D9S1830. We report the first detailed phenotypic study, including neuropsychological, oculographic and brain imaging investigations, in the largest series of AOA2 patients yet recruited. The mean age at onset was 15.1 +/- 3.8 years. Sensory motor neuropathy (92%) and choreic or dystonic movements (44%) were frequent. Oculomotor apraxia was observed in 56% of patients and characterized by increased horizontal saccade latencies and hypometria. AFP levels were elevated in 100% of the families, making it a useful biological marker. This study shows for the first time that AOA2 can be found in Europe, North Africa and the West Indies, and its relative frequency represents approximately 8% of non-Friedreich ARCA, which is more frequent than ataxia telangiectasia and ataxia with oculomotor apraxia type 1 (AOA1), in our series of adult patients. In adults, AOA2 may be, therefore, the most frequent cause of ARCA identified so far, after Friedreich's ataxia. PMID- 14736756 TI - Association between cardiac denervation and parkinsonism caused by alpha synuclein gene triplication. AB - Parkinson's disease patients frequently have symptoms and signs of autonomic nervous dysfunction that are the source of considerable disability. Recent studies have revealed that most patients with Parkinson's disease, and all with Parkinson's disease-associated orthostatic hypotension, have a loss of cardiac sympathetic innervation. Familial Parkinson's disease, caused by mutation of the gene encoding alpha-synuclein, also features orthostatic hypotension, sympathetic neurocirculatory failure and cardiac sympathetic denervation. We have recently described a whole-gene triplication of alpha-synuclein causing Lewy body parkinsonism in a large, well characterized family called the 'Iowa kindred'. Here we report the results of cardiac PET scanning using the sympathoneural imaging agent, 6-[18F]fluorodopamine in affected and unaffected members of this kindred. Four family members were studied, two with parkinsonism, one clinically normal and one with benign essential tremor alone. Both affected members had obvious loss of cardiac sympathetic innervation; the unaffected member had normal innervation, as did the member with isolated essential tremor. The results indicate that, in this family, where disease is caused by overexpression of normal alpha-synuclein, cardiac sympathetic denervation cosegregates with parkinsonism. Post-mortem studies have demonstrated synuclein-positive Lewy body formation in the brains of individuals with parkinsonism who were also in the family described here and who also carry this triplication. These results indicate that both parkinsonism and cardiac sympathetic denervation can result from an excess of normal synuclein. PMID- 14736758 TI - Soccer (football) ocular injuries: an important eye health problem. PMID- 14736759 TI - The nine lives of retinopathy of prematurity. PMID- 14736760 TI - Update of the vascular model of AMD. PMID- 14736761 TI - Distribution and shifting trends of bacterial keratitis in north China (1989-98). AB - AIMS: To study the distribution and shifting trends of bacterial keratitis. METHODS: The data of 2220 corneal isolates from 1 January 1989 to 31 December 1998 were reviewed retrospectively. RESULTS: Positive culture was recovered in 490 isolates. Gram positive cocci and Gram negative bacilli represented 51% and 39.4%, respectively. Pseudomonas aeruginosa was the most common pathogen (32.2%). A gradual increase in the percentage of Gram positive cocci coupled with a decrease of Gram negative bacilli. CONCLUSION: Pseudomonas aeruginosa and coaculase negative Staphylococcus were the most common pathogens in bacterial keratitis in north China. PMID- 14736763 TI - The mechanism and prevention of soccer eye injuries. AB - AIMS: To study the mechanism and the means of preventing soccer eye injuries. METHODS: Kicked soccer ball velocities were measured for a range of ages and experience. Soccer balls (sizes 3, 4, and 5), inflated to 3, 6, and 9 psi, were impacted onto an artificial orbit and the results analysed at 1000 frames per second. Protective eyewear was fitted to a headform then impacted and evaluated. RESULTS: The mean peak ball velocity was 20.4 (SD 6.2) m/s. Soccer balls at 18 m/s entered the orbit between 7.5 and 8.7 mm. There was no significant difference in orbital penetration as a result of ball size or pressure. The soccer ball stayed in the orbit approximately 10 ms and appeared to have a suction effect as it withdrew. Protective eyewear that complied with sports protective eyewear standard ASTM F803 prevented contact of the ball to the eye. CONCLUSIONS: The soccer ball causes eye injury by entering the orbit. Protectors that pass ASTM F803 would prevent orbital intrusion. PMID- 14736764 TI - The effects of single doses of beta radiation on the wound healing behaviour of human Tenon's capsule fibroblasts. AB - AIM: To determine the effects of single doses of beta radiation on the wound healing functions of human Tenon's capsule fibroblasts (hTf). METHODS: hTf were grown in tissue culture and irradiated with beta radiation using a strontium 90 source. The effects of beta radiation on fibroblast migration was studied using microporous transwell membranes. The effects of radiation on fibroblast contraction was investigated using a fibroblast populated collagen gels model. Production of extracellular matrix molecules (collagen I, collagen III, and fibronectin) by monolayers of irradiated fibroblasts was quantified for 14 days following single doses of beta radiation. RESULTS: Growth inhibiting doses of beta radiation did not inhibit fibroblast migration or contraction at any time point. Levels of soluble fibronectin from irradiated populations were significantly reduced after >500 cGy beta radiation. Collagen I and III levels were not reduced after any dose of radiation, and increased following treatment with 1000 cGy beta radiation. CONCLUSIONS: Growth arresting doses of beta radiation have unique effects on the wound healing behaviour of human Tenon's capsule fibroblasts. There was no significant effect on cellular migration or contraction, but ECM production was altered. Fibronectin production was inhibited following higher radiation doses, and collagen I and III production increased after 1000 cGy. The effects of single doses of beta radiation on ocular fibroblast wound healing behaviour are very different from those of 5 fluorouracil and mitomycin C, and these differences may be exploited clinically in the regulation of wound healing after glaucoma filtration surgery. PMID- 14736765 TI - Intraobserver and interobserver reproducibility in the evaluation of ultrasonic pachymetry measurements of central corneal thickness. AB - PURPOSE: To assess reproducibility of central corneal thickness (CCT) measurement by means of ultrasonic pachymetry. METHODS: Fifty one volunteers underwent three sessions of CCT measurements, each consisting of three CCT measurements, performed by each of three different observers. Intra- and interobserver reproducibility was calculated by means of intraclass correlation coefficient (ICC). The expected range of variability between two independent evaluations was calculated using scatter plots of each test-retest difference against their mean. The standard deviation of the mean differences in the test-retest scores was used to describe the differences in the score spread. RESULTS: The ICC ranges of the intra- and interobserver evaluations were 0.95-0.97 and 0.89-0.95 respectively; the expected variability was < or = +/-1% and < or = +/- 2% respectively (95% confidence interval). CONCLUSIONS: The measurement of CCT by means of ultrasonic pachymetry is highly reproducible. PMID- 14736767 TI - Expression of protein gene product 9.5 in lepromatous eyes showing ciliary body nerve damage and a "dying back" phenomenon in the posterior ciliary nerves. AB - BACKGROUND/AIM: Peripheral nerve destruction is the hallmark of leprosy. Ocular complications form a substantial part of the clinical manifestations but histopathology of nerve destruction within ocular structures has not been shown satisfactorily. The role of protein gene product (PGP) 9.5 in identifying nerve destruction in the ciliary body and posterior ciliary nerves of lepromatous eyes is shown. METHODS: Serial sections from two lepromatous eyes and two non lepromatous eyes were stained with PGP 9.5. Histopathological comparison was done on the expression of the PGP 9.5 stain in nerves within the ciliary body, posterior ciliary nerves adjacent to the optic nerve, and nerves tracking through the sclera. RESULTS: In non-lepromatous eyes, PGP 9.5 was expressed in nerves within the ciliary body, the nerves within the sclera, and posterior ciliary nerves adjacent to the optic nerve. In lepromatous eyes no PGP 9.5 was expressed, signifying nerve destruction. CONCLUSIONS: Nerve destruction in lepromatous eyes has been confirmed histopathologically by the absence of or patchy staining with PGP 9.5. Nerve destruction in the ciliary body can extend to the posterior ciliary nerves by an ascending axonopathy. This "dying back" phenomenon is akin to the "glove and stocking" anaesthesia found in lepromatous leprosy. PMID- 14736768 TI - Posterior capsule opacification after implantation of a hydrogel intraocular lens. AB - AIM: To compare the degree of posterior capsule opacification (PCO) in eyes with a hydrophilic hydrogel intraocular lens (IOL) with that in eyes with a hydrophobic acrylic IOL. METHODS: Ninety five patients underwent a hydrogel IOL implantation in one eye and an acrylic IOL implantation in the opposite eye. The PCO value of these patients was measured using the Scheimpflug videophotography system at 1, 6, 12, 18, and 24 months postoperatively. The rate of neodymium:YAG (Nd:YAG) laser posterior capsulotomy and visual acuity were also evaluated. RESULTS: The mean PCO value in the hydrogel group increased significantly (p<0.0001), while that in the acrylic group did not show significant change. The PCO value in the hydrogel group was significantly greater than that in the acrylic group throughout the follow up period. Kaplan-Meier survival analysis determined that the Nd:YAG capsulotomy rate in the hydrogel group was significantly higher than that in the acrylic group (p<0.0001). Mean visual acuity in the hydrogel group decreased significantly with time (p<0.0001), and became significantly worse than that in the acrylic group at 18 and 24 months postoperatively. CONCLUSION: Posterior capsule opacification in eyes with a hydrophilic hydrogel IOL is significantly more extensive than that in eyes with a hydrophobic acrylic IOL, and results in a significant impairment of visual acuity. PMID- 14736769 TI - Case selection in macular relocation surgery for age related macular degeneration. AB - BACKGROUND: To date there has been no randomised controlled trial demonstrating the safety and efficacy of macular relocation surgery (MRS) for age related macular degeneration (AMD). Vision can be improved in some patients and made worse in others despite successful surgery or because of complications. PURPOSE: To determine which patients would benefit from MRS. METHODS: Twenty nine patients with exudative AMD took part in a prospective, non-comparative, interventional study. Macular relocation surgery involved phacoemulsification, vitrectomy, 360 degrees retinotomy, excision of choroidal neovascular membrane, and macular relocation using an infusion of 5-fluorouracil and low molecular weight heparin as adjuvant to prevent proliferative vitreoretinopathy. Patients underwent protocol refraction preoperatively and six-monthly postoperatively by designated optometrists. Preoperative fundus fluorescein angiograms were read by masked observers and the lesions were classified according to a set protocol. The main outcome measures were visual improvement, final vision of better than 20/400, reading speed, critical print size. Logistic and multiple stepwise linear regressions were used to identify independent factors which predicted the main outcomes. RESULTS: Preoperative visual acuity (20/120 or worse) and lesion type (predominantly classic or submacular haemorrhage) were significantly associated with visual improvement (coefficient of regression B = 26.8, p<0.001 and B = 14.9 with p = 0.045 respectively). There were no significant independent factors which predicted a final distance logMAR visual acuity of 1.3 (20/400) or any arbitrary definition of blindness. CONCLUSIONS: The study showed that it was possible to select cases that were more likely to experience an improvement in vision following MRS. PMID- 14736770 TI - Leopard spot retinal pigmentation in infancy indicating a peroxisomal disorder. AB - BACKGROUND: Neonatal adrenoleucodystrophy (NALD) is a rare disorder resulting from abnormal peroxisomal biogenesis. Affected patients present in infancy with developmental delay, hypotonia, and seizures. Blindness and nystagmus are prominent features. The authors suggest a characteristic leopard spot pigmentary pattern in the peripheral retina to be diagnostic. METHODS: Three patients are reported with this presentation; the characteristic retinal appearance resulted in early diagnosis for one of these. CONCLUSION: Leopard spot retinopathy in an infant with hypotonia, seizures, developmental delay, with or without dysmorphic features and hearing impairment, is a clue to the diagnosis of NALD. PMID- 14736771 TI - The prevalence of pseudoexfoliation syndrome in Chinese people. AB - BACKGROUND: Pseudoexfoliation syndrome (PXS) is regarded as rare in people of Chinese ethnicity but the prevalence of this condition is not known. This epidemiology study was conducted to assess the prevalence of PXS in cataract patients and to report the clinical features present. METHODS: Prospective descriptive study conducted in the period from March 1999 to May 2001 in ophthalmology departments in cluster hospitals serving a population of about 1.2 million. 500 consecutive patients aged 60 or above attending the general ophthalmic clinics with a presumed diagnosis of cataract were recruited. A detailed examination including biomicroscopy, intraocular measurement, and gonioscopy were performed on all patients. All positive PXS cases were documented photographically. RESULTS: 500 patients were examined. They ranged from the ages of 60 to 91 years old, with a male to female ratio of 1:2. 40% suffered from hypertension while 24% were known diabetics. Only two positive cases (0.4%) of PXS were identified in the study population. 18% of all eyes were found to have narrow angles (defined as grade 0 to 2 by Shaffer grading). Nuclear sclerosis was the single most common type of lens opacity. CONCLUSION: PXS is a rare condition in Chinese people. A prevalence rate of 0.4% in patients aged 60 or above was identified in this hospital based epidemiology study. To the best of our knowledge, this was the first study conducted in a Chinese population to examine the prevalence of PXS. PMID- 14736772 TI - Do selective topical beta antagonists for glaucoma have respiratory side effects? AB - BACKGROUND/AIMS: Topical beta antagonists are prescribed for glaucoma in approximately 500 000 people in the United Kingdom. The authors have previously shown that topical beta antagonists are associated with an excess incidence of airways obstruction. They examined whether selective topical beta antagonists are associated with excess airways obstruction. METHODS: A historical cohort study was performed to determine the incidence of airways obstruction in subjects with no previous history of airways obstruction, following treatment with topical beta antagonists for glaucoma for the period 1993-7. Cases were defined as having received a first prescription of a drug specifically used in the management of airways obstruction. RESULTS: For selective topical beta antagonists 12 of 324 treated subjects developed airways obstruction, compared with 112 of 9094 controls (adjusted hazard rate 3.0 (95% confidence interval (95% CI) 1.6 to 5.4)). For non-selective topical beta antagonists, the corresponding figures were 69 of 2321 subjects compared with the same control group (adjusted hazard rate 2.2 (1.6 to 3.0)). There was no significant difference between groups (p = 0.47, chi(2)), both being associated with a significantly increased risk of airways obstruction. CONCLUSION: Selective topical beta antagonists do appear to have an excess risk of airways obstruction in this population setting and should be subject to the same prescribing caveats as unselective topical beta antagonists. PMID- 14736774 TI - A 12 week study comparing the fixed combination of latanoprost and timolol with the concomitant use of the individual components in patients with open angle glaucoma and ocular hypertension. AB - BACKGROUND: To compare the intraocular pressure (IOP) lowering effect and safety of the fixed combination of latanoprost and timolol with that of the concomitant use of the individual components. METHODS: A 12 week, double masked, randomised, crossover, multicentre study of patients with open angle glaucoma or ocular hypertension and IOP controlled on ocular hypotensive treatment (mean < or =21 mmHg). Patients received either a once daily morning dose of the fixed combination of latanoprost 0.005% and timolol 0.5% or once daily evening latanoprost 0.005% and twice daily timolol 0.5% for six weeks and then switched to the other combination. The primary efficacy endpoint was the within-patient difference in diurnal IOP between fixed and unfixed treatment combinations after six weeks of treatment; a one sided 97.5% confidence interval (CI) for the mean difference in IOP <1.0 mmHg indicated the fixed combination was not inferior to the unfixed combination. Adverse events were recorded at each visit. RESULTS: In all, 190 patients were included in observed cases analyses (93 fixed to unfixed combination; 97 unfixed to fixed combination). Mean IOP at baseline was 16.9 mmHg in both groups. The mean diurnal IOP was 17.0 mmHg after fixed combination treatment and 15.9 mmHg after unfixed combination therapy (p<0.0001). The difference in mean within-patient diurnal IOP was 1.1 mmHg favouring the unfixed combination (95% CI 0.8 to 1.4 mmHg). Both treatments were tolerated well. CONCLUSIONS: Although the primary efficacy endpoint was not met, once daily administration of the fixed combination of latanoprost and timolol was found to be safe and effective. The fixed combination provides a convenient alternative to the three instillations required with the individual components. PMID- 14736775 TI - The effect of image alignment on capillary blood flow measurement of the neuroretinal rim using the Heidelberg retina flowmeter. AB - AIM: To examine the influence of image alignment on the repeatability of blood flow measurements of the optic nerve. METHODS: 10 normal subjects were examined. Heidelberg retina tomograph imaging was performed to establish best location and focus for the temporal neuroretinal rim. Two high quality Heidelberg retina flowmeter (HRF) images were acquired for three methods of alignment: central, nasal, and temporal. A 10 x 10 pixel measurement window was selected and exactly reproduced on all images. The interquartile pixel values were used to calculate capillary flow. ANOVA, intraclass correlation coefficients (ICC) and the coefficient of repeatability (CoR) were used for analysis. RESULTS: There was no difference between methods (p = 0.47) or between visits (p = 0.51). The ICCs were 0.83 for the central, 0.34 for the nasal, and 0.42 for the temporal alignment. The CoR was 31.5 for central (mean effect 235.1), 234.6 for nasal, and 256.7 for temporal alignment. CONCLUSION: Central alignment was the most repeatable method for the measurement of neuroretinal rim capillary blood flow using the HRF. PMID- 14736776 TI - Clinicopathological findings of choroidal neovascularisation following verteporfin photodynamic therapy. AB - AIMS: To report the clinicopathologic findings of surgically excised choroidal neovascularisation (CNV) three days after verteporfin photodynamic therapy (PDT). METHODS: In three patients (three eyes) with age related macular degeneration, the CNV was surgically removed three days after PDT. The CNV specimens were examined by light microscopy. RESULTS: The patients had subfoveal classic CNV. Fluorescein angiography revealed non-perfusion of the CNV after PDT and before surgery in all eyes. The light microscopy of the CNV membranes showed swollen and damaged endothelium. Thrombus formation or vascular occlusion in the CNV vessels was not detected. CONCLUSION: PDT did not cause a thrombosis of the vessels within the CNV three days after PDT. Severe endothelial damage of the CNV was observed and is likely a primary effect of PDT. Non-perfusion of the CNV at this stage is possibly secondary to occlusion at a deeper level, namely the underlying feeding choroid. PMID- 14736777 TI - Photodynamic therapy with verteporfin is effective, but how big is its effect? Results of a systematic review. AB - BACKGROUND: In 2001 the National Institute for Clinical Excellence (NICE) was asked to issue guidance for England and Wales on the use of photodynamic therapy (PDT). This process has been protracted, partly because of a dispute over the magnitude of beneficial effect. This article examines the origins of the debate about the true treatment effect size for PDT with verteporfin. METHODS: A systematic review of the clinical effectiveness of PDT compared with current practice was undertaken. Searches in Medline, Embase, the Cochrane Library, and the Internet, updated to January 2003, revealed two fully published and four ongoing randomised controlled trials. RESULTS: The results of the two published trials (TAP and VIP) consistently showed that overall, PDT with verteporfin is more effective than placebo in slowing the rate of vision loss. In the TAP trial, 12 or more subgroup analyses were undertaken on the primary outcome measure and in VIP, 10 subgroup analyses but only on a subset of the trial participants. Subgroup analysis results were found to be inconsistent between the two trials, with VIP suggesting that verteporfin was equally effective in occult as in mixed lesions and TAP suggesting that verteporfin was more effective in the predominantly classic subgroup. DISCUSSION: For several reasons it was considered that the most likely estimate of the predominantly classic subgroup effect size was the whole trial result. This has implications for the relationship between cost and benefit, the subject of intense debate. Results of the ongoing trials should help to clarify this subgroup effect size issue. PMID- 14736778 TI - An in vitro study on the compatibility and precipitation of a combination of ciprofloxacin and vancomycin in human vitreous. AB - AIMS: To investigate the precipitation process of a mixture of vancomycin and ciprofloxacin by equilibrium dialysis and its subsequent effect on the level of available free antibiotics. METHODS: Concentrations of vancomycin and ciprofloxacin in an equilibrium dialysis chamber were measured during the equilibrium process by high performance liquid chromatography and fluorescence polarisation immunoassay. Normal saline (NS), balanced salt solution plus (BSS Plus), and vitreous were used separately as the medium of dialysis. RESULTS: Precipitation of ciprofloxacin occurred on incubation at 37 degrees C. It formed precipitate on its own or when mixed with vancomycin in all the three media of NS, BSS Plus, and vitreous. There was more precipitation at higher initial ciprofloxacin concentrations; at 25.0 mg/l about 75% free drug in BSS Plus was lost after 72 hours. The extent of precipitation was similar in both NS and BSS Plus. In the dialysis chambers, 20 mg/l ciprofloxacin dialysed against 125 mg/l vancomycin was reduced to a concentration about 5.0 mg/l after 168 hours. Precipitation of vancomycin was negligible. Ciprofloxacin precipitated in vitreous at body temperature, irrespective of the presence of vancomycin. Even after precipitation, the resultant concentration of ciprofloxacin was still higher than the MIC(90) of the drug against most Gram negative organisms. CONCLUSIONS: Based on this in vitro study, ciprofloxacin precipitated in vitreous at body temperature, irrespective of the presence of vancomycin or the medium for intravitreal injection. The resultant amount of ciprofloxacin was still higher than the MIC(90) of the drug against most Gram negative organisms after precipitation. The authors suggest ciprofloxacin in place of ceftazidime when used in combination with vancomycin for treatment of infective endophthalmitis. PMID- 14736779 TI - Pingelapese achromatopsia: correlation between paradoxical pupillary response and clinical features. AB - AIM: To evaluate the paradoxical pupillary constriction in darkness in patients with Pingelapese achromatopsia (PA), and to describe a connection between this phenomenon and the clinical features. METHODS: 27 patients with PA were examined. All underwent a full ophthalmic examination which included Snellen visual acuity and ophthalmoscopy. Colour vision examination was performed with Ishihara pseudoisochromatic plates and also with a colour plate consisting of five basic colours (red, green, purple, yellow, and orange). Paradoxical pupillary response was examined and documented with a special infrared video camera. Pupils' images were analysed using the Scion Image program and the ratio of pupil size in darkness to its size in light was calculated and recorded. RESULTS: Mean visual acuity was 20/400 (range 20/80-20/800). Colour vision examination showed a mean of 3.2 (SD 1.5) (range 1-5) of Ishihara colour plates, and 0.5 (0.75) (0-3) of basic colour plates. 23 patients (85%) had paradoxical pupillary constriction in darkness. Mean dark/light ratio of pupillary area was 0.86 (range 0.5-1.6). In patients with marked paradoxical pupillary constriction there was a significant correlation of visual acuity and Ishihara score. CONCLUSIONS: Clinical manifestations of achromatopsia include total colour blindness, low visual acuity (mean of 20/400), horizontal pendular or rotatory nystagmus, and photophobia. Most patients have paradoxical pupillary constriction in darkness. When this response is brisk it seems to correlate with lower visual acuity and lower Ishihara score. PMID- 14736780 TI - Multifocal VEP in children: its maturation and clinical application. AB - AIM: To study the maturation of multifocal visual evoked potentials (multifocal VEP) in normal children between the ages of 5 and 16 years and to apply the results clinically in selected cases to the diagnosis of optic pathway diseases. METHOD: 70 normal children were recruited from the community and multifocal VEP (Accumap ObjectiVision, Sydney, Australia) was recorded. The waveform of the evoked responses, the latency and amplitude were analysed. Using these data, an age matched comparison was made with three children with advanced optic nerve disease; two had optic nerve glioma and one had congenital glaucoma. RESULTS: The full field amplitude did not correlate with age and varied greatly within each age group (coefficient of variability 28%). When scaled with respect to the background electroencephalogram the intra-age group variability decreased to 15% and a sigmoid relation was found between amplitude and age. The scaled amplitude remained largely unchanged till 11 years, between 11 and 13 years there was a rapid increase (40%), and remained stable thereafter. This relation was seen at all eccentricities tested. The latency decreased gradually with age and plateaued at 13 years. In the three children with vision abnormalities this test was able to detect scotomas consistent with their condition. CONCLUSION: Multifocal VEP perimetry shows an age related maturation in the visual pathway, characterised by distinctive timeframe of development for amplitude and latency. It can be performed by children as young as 5 years of age and holds promise as a diagnostic test capable of documenting children's visual fields objectively, even before they are able to perform subjective field tests. PMID- 14736781 TI - The Newcastle Control Score: a new method of grading the severity of intermittent distance exotropia. AB - AIM: To describe the development and application of a novel scoring system for grading the severity of intermittent distance exotropia (IDEX) and its potential application as an intervention criterion for surgical intervention. METHODS: The Newcastle Control Score (NCS) for IDEX was developed by incorporating both subjective (home control) and objective (clinic control) criteria into a scale to grade severity. The score structure described was evaluated for interobserver and test-retest reliability. To determine an optimal score threshold for surgical intervention, 170 cases of IDEX were scored retrospectively. Cure rates for surgical and non-surgical cases were then compared according to preoperative or presenting scores. RESULTS: Interobserver and test-test reliability were good (r = 0.82 and r = 0.89 respectively). Total cure rate with surgery was 54% and without surgery 18% (chi(2) = 23.093, df = 1, p<0.001). Significantly fewer patients with NCS >/=3 achieved cure without surgery than those with NCS 2 (chi(2) = 3.362, df = 1, p<0.047). CONCLUSIONS: The NCS is a reliable method for grading the severity of IDEX and aids decisions regarding intervention. Patients with a score of 3 or more are unlikely to attain a cure without surgery. PMID- 14736782 TI - Surgery for primary basal cell carcinoma including the eyelid margins with intraoperative frozen section control: comparative interventional study with a minimum clinical follow up of 5 years. AB - AIM: To assess recurrence of primary basal cell carcinoma (PBCC) including the eyelid margins after resection with or without intraoperative frozen section control (IFS). METHODS: Comparative non-randomised interventional study involving review of records of consecutive patients with histological diagnosis of PBCC including the eyelid margins, treated surgically at the University of Erlangen Nurnberg between 1989 and 1998. Patients with a minimum clinical follow up of 5 years treated with (group I) or without (group II) IFS were compared. Postoperatively, permanent paraffin sections were available in all patients. RESULTS: 165 patients were available for study. There were 145 patients with a minimum of 5 years follow up. Of these, no tumour recurrences were observed in group I (n = 114) compared with three (9.7%) in group II (n = 31) (p = 0.002). CONCLUSIONS: Surgery for PBCC including the eyelid margins with IFS and immediate plastic reconstruction, as performed in the present study, is associated with better long term cure compared with surgery with clinical control. PMID- 14736783 TI - Threshold retinopathy at threshold of viability: the EpiBel study. AB - AIM: To describe incidence, co-morbidity characteristics, and risk factors associated with threshold retinopathy of prematurity (ROP) in survivors with a gestational age (GA) of < or =26 weeks at birth. METHODS: Retrospective analysis of perinatal data of all inborn survivors in all perinatal centres of Belgium in the period 1999-2000 (EpiBel cohort) believed to be between 22 and 26 weeks GA at time of delivery. Data on survivors who did and survivors who did not develop threshold ROP were compared (chi(2), Mann-Whitney U) and logistic regression was performed. RESULTS: Of 303 admitted infants 175 (58%) were discharged alive. Incidence of major retinopathy (> or =stage 3) and of threshold ROP was 25.5% and 19.8% in survivors. Associated central nervous abnormalities were documented in six (17%) and associated chronic lung disease in 19 (54%) threshold ROP infants. Threshold ROP without additional morbidity characteristics at discharge was documented in 14 (40%) infants. Besides often reported risk factors, renal insufficiency (creatinaemia>1.5 mg/dl) was a risk factor to develop threshold ROP (p<0.0015) (chi(2)). Days of respiratory support (OR 1.02; 95% CI 1.002 to 1.039), number of transfusions (OR 1.118; 95% CI 1.030 to 1.214), and renal insufficiency (OR 3.31; 95% CI 1.344 to 8.196) remained independent risk factors to develop threshold ROP in this cohort in a stepwise logistic regression model (MedCalc). CONCLUSIONS: Incidence of threshold ROP is high at the limits of viability. Renal insufficiency is a risk factor to develop threshold ROP in this cohort. PMID- 14736784 TI - Late onset vitreoretinal complications of regressed retinopathy of prematurity. AB - AIM: To report the clinical findings, management, and outcomes in eyes undergoing surgery for regressed retinopathy of prematurity (ROP) with vitreoretinal complications. METHOD: Retrospective review of 40 eyes of 32 patients with regressed ROP who presented between 1989 and 2001 at two UK referral centres. RESULTS: Of 29 eyes presenting with rhegmatogenous retinal detachment (RRD), 15 initially underwent a scleral buckling procedure and 14 initially underwent vitrectomy with or without additional buckling. Primary surgery was anatomically successful in 11/15 eyes that underwent a non-vitrectomy retinal detachment repair and 8/14 that required vitrectomy. The final reattachment rate after reoperation was 28/29 eyes. Median visual acuity improved from 6/60 to 6/36 following retinal detachment repair. A further 11 eyes of eight patients from this series underwent prophylactic surgery, laser, or cryotherapy for predisposing vitreoretinal pathology and/or retinal breaks, all of which were stabilised. CONCLUSIONS: In eyes with RRD and signs of regressed ROP successful reattachment of the retina can be achieved using either vitrectomy or external surgery with an associated overall improvement in visual acuity. A range of external and closed microsurgical approaches is required to effectively deal with the diverse manifestations of regressed ROP. PMID- 14736785 TI - High levels of IgG class antibodies to recombinant HSP60 kDa of Yersinia enterocolitica in sera of patients with uveitis. AB - AIMS: To determine the levels of IgG class antibodies to recombinant heat shock protein 60 kDa of Yersinia enterocolitica (rHSP60Ye), Klebsiella pneumoniae (rHSP60Kp), Escherichia coli (rHSP60Ec), Shigella flexneri (rHSP60Sf), and Streptococcus pyogenes (rHSP60Sp) in the serum of patients with HLA-B27 associated acute anterior uveitis (HLA-B27 associated AAU), idiopathic acute anterior uveitis (idiopathic AAU), pars planitis, Vogt-Koyanagi-Harada (VKH), and healthy subjects. METHODS: The genes that code for HSP60Ye, HSP60Kp, HSP60Ec, HSP60Sf, and HSP60Sp were cloned by PCR from genomic DNA. The rHSPs were purified by affinity using a Ni-NTA resin. The serum levels of IgG class antibodies to rHSP60s were determined by ELISA in patients with uveitis (n = 42) and in healthy subjects (n = 25). RESULTS: The majority of patients with uveitis had higher levels of IgG class antibodies to rHSP60Ye compared with levels of healthy subjects (p = 0.01), although these differences were only observed in the HLA-B27 associated AAU (p = 0.005) and in pars planitis patients (p = 0.001). The levels of IgG antibodies to the rHSP60Kp, rHSP60Sf, rHSP60Ec, and rHSP60Sp were similar in patients with uveitis and in healthy subjects (p>0.05). CONCLUSION: The results suggest that HSP60Ye could be involved in the aetiology of HLA-B27 associated AAU and pars planitis. PMID- 14736786 TI - Alterations in the morphology of lamina cribrosa pores in glaucomatous eyes. AB - AIMS: To determine alterations which occur in the size and shape of lamina cribrosa (LC) pores in glaucomatous eyes over a period of time. METHODS: Baseline and follow up optic disc photographs were retrospectively studied in 39 eyes of 39 patients with glaucoma. Only eyes with a vertical cup to disc ratio equal to or greater than 0.6 were included in the study. In addition, all selected eyes had to have serial optic disc photographs obtained at least 3 years apart allowing clear visualisation of LC surface. The association of the alterations in LC surface morphology with patient specific and eye specific characteristics was statistically analysed. RESULTS: During a mean study period of 3.90 (SD 0.7) years, individual pore size (mean pore area to disc area ratio) exhibited a significant decrease between baseline and follow up measurements of each eye (p<0.0001). However, during the study period, total pore area to disc area ratio did not change (p>0.05), and the change in pore shape in some eyes (from circular to more oval and elongated) was statistically insignificant (p = 0.12). Although a relation was detectable between the optic disc and lamina cribrosa parameters at a given time, which reflects cumulative effects, during the study period, there was no significant association between the changes of the LC parameters and neural tissue damage. The rate and the magnitude of the changes in individual pore size during the study period were not significantly different among the eyes exhibiting progressive neural rim damage and those staying stable (p>0.05). CONCLUSION: These findings demonstrate that the LC surface morphology exhibits changes along with the glaucomatous optic disc damage. However, the clinical appearance of LC surface in glaucomatous eyes may continue to change, even when the neural rim damage is clinically stable. These findings are probably associated with the chronic cellular events of tissue remodelling that occur in the glaucomatous optic nerve head. PMID- 14736787 TI - Effects of brinzolamide on ocular haemodynamics in healthy volunteers. AB - AIM: A prospective, randomised study to evaluate effects of brinzolamide on ocular haemodynamics in healthy volunteers. METHODS: 30 volunteers (12 men, 18 women; 28.3 (SD 7.8) years) were prospectively randomised to either brinzolamide or placebo during a 2 week double masked treatment trial. Examinations were performed at baseline and after 2 weeks of treatment. Intraocular pressure was measured and automatic static perimetry (Humphrey field analyser, 24-2) and contrast sensitivity (CSV 1000, Vector Vision) were performed. Retrobulbar blood flow velocities (peak systolic and end diastolic velocity) and resistive indices (RI) of ophthalmic artery, central retinal artery and of temporal and nasal short posterior ciliary arteries were measured by colour Doppler imaging (Sonoline Sienna Siemens). In video fluorescein angiograms (scanning laser ophthalmoscope, Rodenstock) arteriovenous passage time (AVP, dilution curves) and peripapillary diameters of retinal arterioles and venules were measured by means of digital image analysis. RESULTS: Intraocular pressure was significantly decreased by brinzolamide (p<0.0001). Neither brinzolamide nor placebo changed visual field global indices after treatment. Contrast sensitivity at 3 cycles per degree was significantly higher in the placebo group (p<0.05). Apart from an increase of RI in ophthalmic artery under placebo treatment (p<0.05) there was no effect in retrobulbar haemodynamics in both groups. Brinzolamide therapy alone resulted in a significant reduction of AVP compared to baseline (p<0.05), while peripapillary retinal vessels diameters remained unaffected. CONCLUSIONS: Apart from the expected decrease of intraocular pressure brinzolamide showed no significant change in retrobulbar haemodynamics, but a significant shortening of AVP. Since in glaucoma AVP is prolonged indicating vascular dysfunction this effect might be beneficial in glaucoma therapy. PMID- 14736788 TI - ARIX gene polymorphisms in patients with congenital superior oblique muscle palsy. AB - AIM: To identify ARIX gene polymorphisms in patients with congenital superior oblique muscle palsy and to find the relation between the ARIX gene and congenital superior oblique muscle palsy. METHODS: The three exons of the ARIX gene were sequenced by genomic DNA amplification with polymerase chain reaction (PCR) and direct sequencing in 15 patients with superior oblique muscle palsy (13 with congenital and two with acquired palsy) and 54 normal individuals. PCR products cloned into plasmids were also sequenced. A family with father and a daughter each having congenital superior oblique muscle palsy was also involved in this study. RESULTS: Four patients with congenital superior oblique muscle palsy carried heterozygous nucleotide changes in the ARIX gene. One patient with the absence of the superior oblique muscle had T7C in the 5'-UTR of the exon 1 and C-44A in the promoter region, both of which were located on the same strand. Another unrelated patient with congenital superior oblique muscle palsy had C76G in the 5'-UTR of the exon 1 and C-9A in the promoter region on the same strand. G153A in the 5'-UTR of exon 1 was found in common in two affected members of a family with congenital superior oblique muscle palsy. This G153A in the 5'-UTR of exon 1 was also present in four unrelated normal individuals. No other heterozygous nucleotide changes were found in normal individuals. CONCLUSIONS: The nucleotide change (G153A) in the 5'-UTR of exon 1 co-segregated with congenital superior oblique muscle palsy in one family. Four other nucleotide changes in the exon 1 or the promoter region were found only in patients with congenital superior oblique muscle palsy. These nucleotide polymorphisms may be one of the risk factors for the development of congenital superior oblique muscle palsy. PMID- 14736789 TI - Unexplained head tilt following surgical treatment of congenital esotropia: a postural manifestation of dissociated vertical divergence. AB - BACKGROUND: Strabismus surgery for congenital esotropia can be complicated by the development of a postoperative head tilt. PURPOSE: To determine the pathophysiology of acquired head tilting following horizontal realignment of the eyes in children with congenital esotropia. MATERIALS AND METHODS: Retrospective analysis of nine children with congenital esotropia who developed unexplained head tilts following horizontal realignment of the eyes. RESULTS: Shortly after strabismus surgery, each child developed a head tilt in association with asymmetrical dissociated vertical divergence (DVD). Five children maintained a head tilt toward the side of the fixing eye (group 1), which did not serve to control the DVD. Four children maintained a head tilt toward the side of the hyperdeviating eye, which served to control the DVD (group 2). Children in group 2 had earlier horizontal muscle surgery and developed better stereopsis than those in group 1, suggesting that the higher degree of single binocular vision and stereopsis in these children may have led to a compensatory torticollis to control an asymmetrical DVD. CONCLUSIONS: The onset of an unexpected head tilt after congenital esotropia surgery is usually a postural manifestation of asymmetrical DVD. In this setting, a head tilt toward the side of the fixing eye corresponds with a postural manifestation of the underlying central vestibular imbalance that produces DVD, while a head tilt toward the side of the hyperdeviating eye serves to counteract the hyperdeviation and stabilise binocular vision. PMID- 14736790 TI - Susceptibility to endotoxin induced uveitis is not reduced in mice deficient in BLT1, the high affinity leukotriene B4 receptor. AB - AIM: To investigate the role of arachidonic acid derived chemotactic factor, LTB(4), in the development of endotoxin induced uveitis (EIU), using mice deficient in the BLT1 gene which encodes the high affinity LTB(4) receptor. METHODS: BLT1 gene deficient and wild type BALB/c mice were injected intravitreally with Escherichia coli 055:B5 lipopolysaccharide (250 ng/2 microl). Number of leukocytes invading the anterior chamber 24 hours later were counted on tissue cross sections. RESULTS: In all mice, EIU was characterised by a polymorphonuclear and mononuclear cell infiltrate. Numbers of infiltrating cells did not differ significantly between control and BLT1 gene knockout mice. CONCLUSION: Chemotactic factors other than LTB(4) are primarily responsible for leukocyte migration into the eye during murine EIU. PMID- 14736791 TI - Optic nerve oxygen tension: the effects of timolol and dorzolamide. AB - BACKGROUND/AIMS: The authors have previously reported that carbonic anhydrase inhibitors such as acetazolamide and dorzolamide raise optic nerve oxygen tension (ONPO(2)) in pigs. The purpose of the present study was to investigate whether timolol, which belongs to another group of glaucoma drugs called beta blockers, has a similar effect. In addition, the effect of dorzolamide and timolol in combination was studied. METHODS: Polarographic oxygen electrodes were placed transvitreally over the optic disc in anaesthetised pigs and ONPO(2) was recorded continually. Drugs were administered intravenously either as 100 mg timolol followed by 500 mg dorzolamide (n = 5), 500 mg dorzolamide followed by 100 mg timolol (n = 5), or 100 mg timolol and 500 mg dorzolamide given simultaneously (n = 5). Arterial blood pressure, blood gasses, and heart rate were recorded. RESULTS: ONPO(2) was unaffected by administration of 100 mg timolol as an intravenous injection (n = 5). Administration of 500 mg dorzolamide by itself significantly increased ONPO(2) from 2.96 (SD 0.62) kPa to 3.69 (SD 0.88) kPa (n = 4, p = 0.035). The dorzolamide induced ONPO(2) increase was not significantly different from the ONPO(2) increases were seen when dorzolamide was administered simultaneous with (n = 5) or 35 minutes (n = 5) after 100 mg timolol. CONCLUSION: Systemic administration of timolol does not affect the optic nerve oxygen tension despite its lowering effect on the intraocular pressure. Additionally, timolol does not affect the ONPO(2) increasing effect of dorzolamide. PMID- 14736792 TI - Ocular surface epithelium induces expression of human mucosal lymphocyte antigen (HML-1) on peripheral blood lymphocytes. AB - BACKGROUND/AIMS: Peripheral blood CD8+ lymphocytes that home to mucosal surfaces express the human mucosal lymphocyte antigen (HML-1). At mucosal surfaces, including the ocular surface, only intraepithelial CD8+ lymphocytes express HML 1. These lymphocytes are retained in the intraepithelial compartment by virtue of the interaction between HML-1 and its natural ligand, E-cadherin, which is expressed on epithelial cells. The purpose of this study was to determine whether ocular surface epithelial cells (ocular mucosa) could induce the expression of human mucosal lymphocyte antigen on peripheral blood lymphocytes. METHODS: Human corneal and conjunctival epithelial cells were co-cultured with peripheral blood lymphocytes. Both non-activated and activated lymphocytes were used in the experiments. After 7 days of incubation, lymphocytes were recovered and analysed for the antigens CD8/HML-1, CD4/HML-1, CD3/CD8, CD3/CD4, CD3/CD25, CD8/CD25, and CD4/CD25 by flowcytometry. RESULTS: Significant statistical differences were observed in the CD8/HML-1 expression when conjunctival epithelial cells were co cultured with non-activated and activated lymphocytes (p = 0.04 for each) and when corneal epithelial cells were co-cultured with non-activated lymphocytes (p = 0.03). Significant statistical difference in CD4/HML-1 expression was observed only when conjunctival epithelial cells were co-cultured with activated lymphocytes (p = 0.02). CONCLUSION: Ocular surface epithelial cells can induce the expression of human mucosal lymphocyte antigen on CD8+ (and to some extent on CD4+) lymphocytes. This may allow the retention of CD8+ and CD4+ lymphocytes within the epithelial compartment of the conjunctiva and play a part in mucosal homing of lymphocytes. PMID- 14736793 TI - The distributions of mitochondria and sodium channels reflect the specific energy requirements and conduction properties of the human optic nerve head. AB - AIM: To study the normal distributions of mitochondria and voltage gated Na+ channels in the human optic nerve head in order to gain insight into the potential mechanisms of optic nerve dysfunction seen in the inherited optic neuropathies. METHODS: Five fresh frozen human optic nerves were studied. Longitudinally orientated, serial cryosections of optic nerve head were cut for mitochondrial enzyme histochemistry and immunolabelling for cytochrome c oxidase (COX) subunits and voltage gated Na+ channel subtypes (Na(v) 1.1, 1.2, 1.3, and 1.6). RESULTS: A high density of voltage gated Na+ channels (subtypes Na(v) 1.1, 1.3, and 1.6) in the unmyelinated, prelaminar, and laminar optic nerve was found. This distribution co-localised both with areas of high COX activity and strong immunolabelling for COX subunits I and IV. CONCLUSIONS: Increased numbers of mitochondria in the prelaminar optic nerve have previously been interpreted as indicating a mechanical hold up of axoplasmic flow at the lamina cribrosa. These results suggest that this increased mitochondrial density serves the higher energy requirements for electrical conduction in unmyelinated axons in the prelaminar and laminar optic nerve and is not a reflection of any mechanical restriction. This could explain why optic neuropathies typically occur in primary inherited mitochondrial diseases such as Leber's hereditary optic neuropathy, myoclonic epilepsy with ragged red fibres (MERRF), and Leigh's syndrome. Secondary mitochondrial dysfunction has also been reported in dominant optic atrophy, Friedreich's ataxia, tobacco alcohol amblyopia, Cuban epidemic optic neuropathy, and chloramphenicol optic neuropathy. These diseases are rare but these findings challenge the traditional theories of optic nerve structure and function and may suggest an alternative approach to the study of commoner optic neuropathies such as glaucoma. PMID- 14736795 TI - Heparin therapy in giant cell arteritis. PMID- 14736794 TI - The cone dysfunction syndromes. AB - The cone dystrophies comprise a heterogeneous group of disorders characterised by visual loss, abnormalities of colour vision, central scotomata, and a variable degree of nystagmus and photophobia. They may be stationary or progressive. The stationary cone dystrophies are better described as cone dysfunction syndromes since a dystrophy often describes a progressive process. These different syndromes encompass a wide range of clinical and psychophysical findings. The aim is to review current knowledge relating to the cone dysfunction syndromes, with discussion of the various phenotypes, the currently mapped genes, and genotype phenotype relations. The cone dysfunction syndromes that will be discussed are complete and incomplete achromatopsia, oligocone trichromacy, cone monochromatism, blue cone monochromatism, and Bornholm eye disease. Disorders with a progressive cone dystrophy phenotype will not be discussed. PMID- 14736796 TI - Retinal migraine: caught in the act. PMID- 14736797 TI - Long term follow up in a case of successfully treated idiopathic retinal vasculitis, aneurysms, and neuroretinitis (IRVAN). PMID- 14736798 TI - HLA typing is not predictive of proliferative diabetic retinopathy in patients with younger onset type 2 diabetes mellitus. PMID- 14736799 TI - Effect of docosahexaenoic acid supplementation on retinal function in a patient with autosomal dominant Stargardt-like retinal dystrophy. PMID- 14736800 TI - Severe retinopathy of prematurity (ROP) in a premature baby treated with sildenafil acetate (Viagra) for pulmonary hypertension. PMID- 14736801 TI - Indocyanine green localisation in surgically excised choroidal neovascular membrane in age related macular degeneration. PMID- 14736802 TI - Dopamine is an indicator but not an independent risk factor for grade 3 retinopathy of prematurity in extreme low birthweight infants. PMID- 14736803 TI - Correction of pseudophakic anisometropia in a patient with pseudoexfoliation using an implantable contact lens. PMID- 14736804 TI - Consent of the blind and visually impaired: a time to change practice. PMID- 14736805 TI - Magnetic resonance angiography source images in carotid cavernous fistulas. PMID- 14736806 TI - Are we overlooking the side effects of the drugs in our zeal to conquer ARMD? PMID- 14736807 TI - Response to allegations and some considerations on interferon treatment in Behcet's disease. PMID- 14736810 TI - Male reproductive toxicity of trichloroethylene: sperm protein oxidation and decreased fertilizing ability. AB - The objective of the present study was to characterize and investigate potential mechanisms for the male reproductive toxicity of trichloroethylene (TCE). Male rats exposed to TCE in drinking water exhibited a dose-dependent decrease in the ability to fertilize oocytes from untreated females. This reduction in fertilizing ability occurred in the absence of treatment-related changes in combined testes/epididymides weight, sperm concentration, or sperm motility. In addition, flow cytometric analysis showed that there were no treatment-related differences in sperm mitochondrial membrane potential or acrosomal stability. TCE caused slight histological changes in efferent ductule epithelium, coinciding with the previously reported ductule localization of cytochrome P450 2E1. However, no alterations were noted in the testis or in any segment of the epididymis. Because there were no treatment-related changes to sperm indices and no clear pathological lesions to explain the reduced fertilization, the present study investigated TCE-mediated sperm oxidative damage. Oxidized proteins were detected by immunochemical techniques following the derivatization of sperm protein carbonyls with dinitrophenyl hydrazine. Immunochemical staining of whole, intact sperm showed the presence of halos of oxidized proteins around the head and midpiece of sperm from TCE-treated animals. The presence of oxidized sperm proteins was confirmed by Western blotting using in vitro-oxidized sperm as a positive control. Thiobarbituric acid reactive substances analyses showed a dose dependent increase in the level of lipid peroxidation in sperm from treated animals, as well. Oxidative damage to sperm may explain the diminished fertilizing capacity of exposed animals and provide another mechanism by which TCE can adversely affect reproductive capabilities in the male. PMID- 14736811 TI - European hamsters (Cricetus cricetus) show a transient phase of insensitivity to long photoperiods after gonadal regression. AB - Annual rhythms of body weight and reproduction in the European hamster (Cricetus cricetus) are the result of an interaction between seasonal changes in day length (photoperiod) and seasonal changes in the responsiveness of animals to these photoperiods. The present study demonstrates that under natural conditions European hamsters are not able to perceive long photoperiods (i.e., a 16L:8D cycle) before mid-November. This is an important difference to other hamster species, in which regrowth of the gonads can be stimulated by exposure to long photoperiods at any stage of gonadal regression. The experiments also demonstrate the existence of an annual phase of sensitivity to long photoperiods that starts around mid-November and extends until March/April. During this phase of sensitivity, exposure to a long photoperiod (16L:8D) induced gonadal regrowth within 3 wk. Additional experiments with an accelerated photoperiodic lighting regimen indicated that a photoperiod of approximately 13 h is necessary to stimulate gonadal regrowth. Under natural light conditions in Stuttgart (48.46 degrees N), a photoperiod of 13 h is reached by the beginning of April, which fits well with the finding that the majority of animals kept under a natural light:dark cycle had well-developed gonads by the end of April. Nevertheless, these animals showed a rather variable timing of gonadal regrowth, ranging from early January to late April. This is most likely the result of two processes: first, an endogenous mechanism (photorefractoriness) that induces gonadal recrudescence without any photoperiodic information while the animals are still in their hibernation burrows, and second, a direct stimulatory effect of long photoperiods. PMID- 14736812 TI - Embryonic stem cells expressing both platelet endothelial cell adhesion molecule 1 and stage-specific embryonic antigen-1 differentiate predominantly into epiblast cells in a chimeric embryo. AB - We examined the expression of cell-surface markers on subpopulations of mouse embryonic stem (ES) cells to identify those that were associated with cells that had the highest pluripotency. Flow cytometry analysis revealed a wide variation in the expression of platelet endothelial cell adhesion molecule 1 (PECAM-1) and stage-specific embryonic antigen (SSEA)-1 in ES cells. Almost all SSEA-1+ cells expressed a high level of PECAM- 1, and reversible repopulation was observed between PECAM- 1+SSEA-1+ and PECAM-1+SSEA-1- cells. The ES cells carrying the lacZ gene were sorted into three subpopulations: PECAM- 1-SSEA-1-, PECAM-1+SSEA-1 , and PECAM-1+SSEA-1+. Quantitative reverse transcription-polymerase chain reaction revealed a low level of Oct3/4 mRNA expression and an elevation in differentiation maker gene expression in PECAM-1- cells. To compare the pluripotency of these three subpopulations, a single cell from each was injected into eight-cell embryo and ES cells identified at later stages by X-gal staining. At the blastocyst stage, PECAM-1+ SSEA-1+/- cells were found to have differentiated into epiblast cells in high numbers. In contrast, PECAM- 1- cell derivatives localized in the primitive endoderm or trophectoderm. At 6.0-7.0 days post coitum, many PECAM-1+SSEA- 1+ cells were found in the epiblast, but few beta gal+ cells were detected in any regions of embryos that were injected with cells from the other two populations. These results showed that the expression levels of PECAM-1 and SSEA-1 in ES cells correlated closely with their pluripotency and/or their ability to incorporate into the epiblast of chimeric embryos. PMID- 14736813 TI - Activin A and follicle-stimulating hormone control tight junctions in avian granulosa cells by regulating occludin expression. AB - Within the avian ovarian follicle, the oocyte is surrounded by a monolayer of granulosa cells, which exhibit pronounced epithelial properties. Here we demonstrate the presence of the major tight junction protein occludin in granulosa cells. As shown by immunohistochemistry, occludin localizes to the oocyte-facing granulosa cell surface. Occludin and thus tight junctions are dynamically regulated in a developmental stage-specific manner. Small white follicles, which have not yet started yellow yolk incorporation, show pronounced occludin expression in vitro and in vivo. By contrast, yellow yolk-incorporating small yellow follicles exhibit much lower levels of occludin, and hierarchical, preovulatory follicles are virtually devoid of this essential tight junction component. Using a primary granulosa cell culture system, we demonstrate that concerted action of two well-established ovarian growth regulators, follicle stimulating hormone and activin A, leads to strong induction of occludin expression in vitro. We suggest that the stage-dependent decrease in the granulosa cell growth factor responsiveness triggers the disruption of tight junctions, enabling rapid and high capacity transport of macromolecules into the oocyte through a paracellular pathway. Such a high-capacity transport for yolk components may represent a crucial prerequisite for rapid oocyte growth once follicles have entered the follicular hierarchy. PMID- 14736814 TI - Estrogenic activities of nitrophenols in diesel exhaust particles. AB - We recently isolated 3-methyl-4-nitrophenol (4-nitro-m-cresol; PNMC) and 4-nitro 3-phenylphenol (PNMPP) from diesel exhaust particles (DEP) and identified them as vasodilators. Because these compounds are alkylphenolic derivatives that might mimic hormones, we evaluated their estrogenic activity by using recombinant yeast screens, myometrial contractility assays, and in vivo uterotrophic assays. Recombinant yeast screen assays showed that both PNMC and PNMPP possess estrogenic activity. Furthermore, ovariectomized 25-day-old immature female rats injected with PNMC and PNMPP subcutaneously for 2 days showed significant increases in uterine weight among those receiving 100 mg/kg PNMC and 0.1 and 1.0 mg/kg PNMPP. To clarify further the estrogenic activity of PNMC and PNMPP, rat uterine horns were monitored in organ bath chambers for myometrial contractility in response to oxytocin (OT). Significant differences occurred in the initial and maximum contractilities to OT at 0.25 and 25 mIU/ml in uterine horns obtained from animals treated with 100 mg/kg PNMC and in the maximum contractilities to OT at 0.025, 0.25, and 25 mIU/ml in those from rats treated with 0.1 mg/kg PNMPP. These results clearly demonstrated that PNMC and PNMPP in DEP have estrogenic activity both in vitro and in vivo and might therefore be considered as endocrine disrupting chemicals. PMID- 14736815 TI - Identification of genes involved in apoptosis and dominant follicle development during follicular waves in cattle. AB - We hypothesize that granulosa and theca cells from growing dominant follicles, with relatively high intrafollicular concentrations of estradiol, have a greater expression of genes involved in inhibiting apoptosis pathways and lower expression of genes involved in apoptosis pathways than growing subordinate follicles with lower estradiol concentrations. Using the well-characterized bovine dominant follicle model, we collected granulosa and theca cells from individual dominant and the largest subordinate follicle 3 days after initiation of a follicular wave in four animals. Based on ultrasound analysis, both follicle types were in the growth phase at the time of ovariectomy. However, dominant follicles were larger (9.8 +/- 1.0 versus 7.6 +/- 0.6 mm in diameter, P < 0.05) and had greater intrafollicular concentrations of estradiol (132.2 +/-3 8.5 versus 24.1 +/- 12.1 ng/ml, P < 0.05), compared with the largest subordinate follicles. We used bovine cDNA microarrays, which contained a total of 1400 genes, including a subset of 53 genes known to be involved in apoptosis pathways, to determine which apoptosis and marker genes from each of the four dominant versus subordinate follicles were potentially differentially expressed. Using a low stringency-screening criterion, 22 genes were identified. Quantitative real time polymerase chain reaction confirmed that 16 of these genes were differentially expressed. Our novel results demonstrate that the high intrafollicular concentrations of estradiol in growing dominant follicles were positively associated with enhanced expression of mRNAs in granulosa cells for aromatase, LH receptor, estradiol receptor beta, DICE-1, and MCL-1, compared with granulosa cells from subordinate follicles (all survival-associated genes). In contrast, the relatively low intrafollicular concentrations of estradiol in growing subordinate follicles were positively associated with enhanced expression of mRNAs in granulosa cells for beta glycan, cyclo-oxygenase-1, tumor necrosis factor alpha, caspase-activated DNase, and DRAK-2, and in theca cells for beta glycan, caspase 13, P58(IPK), Apaf-1, BTG-3, and TS-BCLL, compared with granulosa or theca cells from dominant follicles (genes that are all associated with cell death and/or apoptosis). We suggest that that these genes may be candidate estradiol target genes and that they may be early markers for the final stages of follicle differentiation or initiation of apoptosis and thus selection of dominant follicles during follicular waves. PMID- 14736816 TI - Variation in the membrane transport properties and predicted optimal rates of freezing for spermatozoa of diploid and tetraploid Pacific oyster, Crassostrea gigas. AB - In the present study, a shape-independent differential scanning calorimeter (DSC) technique was used to measure the dehydration response during freezing of sperm cells from diploid and tetraploid Pacific oysters, Crassostrea gigas. This represents the first application of the DSC technique to sperm cells from nonmammalian species. Volumetric shrinkage during freezing of oyster sperm cell suspensions was obtained at cooling rates of 5 and 20 degrees C/min in the presence of extracellular ice and 8% (v/v) concentration of dimethyl sulfoxide (DMSO), a commonly used cryoprotective agent (CPA). Using previously published data, sperm cells from diploid oysters were modeled as a two-compartment "ball-on stick" model with a "ball" 1.66 microm in diameter and a "stick" 41 microm in length and 0.14 microm wide. Similarly, sperm cells of tetraploid oysters were modeled with a "ball" 2.14 microm in diameter and a "stick" 53 microm in length and 0.17 microm wide. Sperm cells of both ploidy levels were assumed to have an osmotically inactive cell volume, Vb, of 0.6 Vo, where Vo is the isotonic (or initial) cell volume. By fitting a model of water transport to the experimentally obtained volumetric shrinkage data, the best-fit membrane permeability parameters (Lpg and ELp) were determined. The combined-best-fit membrane permeability parameters at 5 and 20 degrees C/min for haploid sperm cells (or cells from diploid Pacific oysters) in the absence of CPAs were: Lpg = 0.30 x 10(-15) m(3)/Ns (0.0017 microm/min-atm) and ELp = 41.0 kJ/mole (9.8 kcal/mole). The corresponding parameters in the presence of 8% DMSO were: Lpg[cpa] = 0.27 x 10( 15) m(3)/Ns (0.0015 microm/min-atm) and ELp[cpa] = 38.0 kJ/mole (9.1 kcal/mole). Similarly, the combined-best-fit membrane permeability parameters at 5 and 20 degrees C/min for diploid sperm cells (or cells from tetraploid Pacific oysters) in the absence of CPAs were: Lpg = 0.34 x 10(-15) m(3)/Ns (0.0019 microm/min-atm) and ELp = 29.7 kJ/mole (7.1 kcal/mole). The corresponding parameters in the presence of 8% DMSO were: Lpg[cpa] = 0.34 x 10(-15) m(3)/Ns (0.0019 microm/min atm) and ELp[cpa] = 37.6 kJ/mole (9.0 kcal/mole). The parameters obtained in this study suggest that optimal rates of cooling for Pacific oyster sperm cells range from 40 to 70 degrees C/min. These theoretical cooling rates are in close conformity with empirically determined optimal rates of cooling sperm cells from Pacific oysters, C. gigas. PMID- 14736817 TI - Glutamine synthesis in the developing porcine placenta. AB - Glutamine plays a vital role in fetal carbon and nitrogen metabolism and exhibits the highest fetal:maternal plasma ratio among all amino acids in pigs. Such disparate glutamine levels between mother and fetus suggest that glutamine may be actively synthesized and released into the fetal circulation by the porcine placenta. We hypothesized that branched-chain amino acid (BCAA) metabolism in the placenta plays an important role in placental glutamine synthesis. This hypothesis was tested by studying conceptuses from gilts on Days 20, 30, 35, 40, 45, 50, 60, 90, or 110 of gestation (n = 6 per day). Placental tissue was analyzed for amino acid concentrations, BCAA transport, BCAA degradation, and glutamine synthesis as well as the activities of related enzymes (including BCAA transaminase, branched-chain alpha-ketoacid dehydrogenase, glutamine synthetase, glutamate-pyruvate transaminase, and glutaminase). On all days of gestation, rates of BCAA transamination were much greater than rates of branched-chain alpha ketoacid decarboxylation. The glutamate generated from BCAA transamination was primarily directed to glutamine synthesis and, to a much lesser extent, alanine production. Placental BCAA transport, BCAA transamination, glutamine synthesis, and activities of related enzymes increased markedly between Days 20 and 40 of gestation, as did glutamine in fetal allantoic fluid. Accordingly, placental BCAA levels decreased after Day 20 of gestation in association with a marked increase in BCAA catabolism and concentrations of glutamine. There was no detectable catabolism of glutamine in pig placenta throughout pregnancy, which would ensure maximum output of glutamine by this tissue. These novel results demonstrate glutamine synthesis from BCAAs in pig placentae, aid in explaining the abundance of glutamine in the fetus, and provide valuable insight into the dynamic role of the placenta in fetal metabolism and nutrition. PMID- 14736818 TI - Accelerated maturation of primate testis by xenografting into mice. AB - Testicular maturation and sperm production throughout the life of the male form the basis of male fertility. It is difficult to elucidate the intricate processes controlling testicular maturation and spermatogenesis in primates in vivo due to the long time span required for sexual maturation and also to the lack of accessible in vitro or in vivo models of primate spermatogenesis. Ectopic xenografting of neonatal testis tissue into mice provides an accessible model to study and manipulate the propagation and differentiation of male germ cells from immature donor animals. However, it was not clear whether this approach would be applicable to slowly maturing primates. Here we report that grafting of testis tissue from immature rhesus monkeys (Macaca mulatta) into host mice resulted in the acceleration of testicular maturation and production of fertilization competent sperm in testis xenografts. The system reported here provides a powerful, practical approach to study timing and control of testicular maturation and regulation of primate spermatogenesis without the necessity for experimentation in primates. This approach could potentially be applied to produce fertile sperm from sexually immature individuals of rare or valuable primate species or from prepubertal boys undergoing sterilizing therapy for cancer. PMID- 14736819 TI - Meiosis-activating sterol promotes the metaphase I to metaphase II transition and preimplantation developmental competence of mouse oocytes maturing in vitro. AB - The objective of this study was to determine the effects of a sterol found in ovarian follicular fluid, known as meiosis-activating sterol (FF-MAS), on the maturation of mouse oocytes in vitro. Possible effects of FF-MAS in promoting the metaphase I (MI) to metaphase II (MII) transition (nuclear maturation) and the competence of oocytes to complete preimplantation embryo development to the blastocyst stage (cytoplasmic maturation) were assessed. Cumulus cell-enclosed oocytes that were compromised in their ability to undergo nuclear maturation and subsequent development because of the age or genotype of the female were isolated at the germinal vesicle stage and matured in vitro using media supplemented with 0 to 20 microM FF-MAS. Oocytes that progressed to MII were inseminated in vitro, and the percentages developing to the 2-cell and blastocyst stages were determined. The sterol was omitted from the media used for oocyte insemination or preimplantation development. FF-MAS promoted a significantly higher percentage of oocytes in all groups to progress to MII in vitro. Moreover, FF-MAS treatment of oocytes maturing in vitro dramatically increased the competence of all but one of the groups of oocytes to complete preimplantation development. Therefore, FF-MAS improved mouse oocyte quality by promoting both nuclear and cytoplasmic maturation in vitro. PMID- 14736820 TI - Bayesian analysis and risk assessment in genetic counseling and testing. AB - Risk assessment is an essential component of genetic counseling and testing, and Bayesian analysis plays a central role in genetic risk assessment. Bayesian analysis allows calculation of the probability of a particular hypothesis, either disease or carrier status, based on family information and/or genetic test results. Genetic risk should be assessed as accurately as possible for family decision making. Additional information, from the pedigree and/or from genetic testing, can often dramatically improve the accuracy of genetic risk assessment. We illustrate herein the application of Bayes' theorem and describe important basic principles in genetic risk assessment. PMID- 14736821 TI - Chromogenic in situ hybridization for alpha6beta4 integrin in breast cancer: correlation with protein expression. AB - The alpha6beta4 integrin is the receptor for the basement membrane protein laminin-5. Recent studies suggest that alpha6beta4 integrin expression in invasive breast carcinomas may be a poor prognostic factor. Because we have not had reliable results with commercially available antibodies for the immunohistochemical detection of alpha6beta4 integrin in archival paraffin embedded tissues, we designed a probe to detect beta4 integrin subunit mRNA in paraffin sections. In situ hybridization for beta4 mRNA was performed on paraffin embedded tissue sections of 25 invasive breast carcinomas using a hyperbiotinylated oligonucleotide DNA probe. Immunohistochemical staining was performed on corresponding frozen tumor sections using two commercially available antibodies to the beta4 integrin subunit. All cases positive for beta4 protein by one or both antibodies were also positive for beta4 mRNA by in situ hybridization, but three cases with beta4 mRNA expression were negative by immunohistochemistry with both antibodies. These findings suggest that in situ hybridization appears to be a sensitive method for detecting beta4 integrin mRNA, but it appears to identify some cases that either lack beta4 protein or express variants not recognized with commercial antibodies directed to particular extracellular or cytoplasmic domains. PMID- 14736822 TI - Utility of linearly amplified RNA for RT-PCR detection of chromosomal translocations: validation using the t(2;5)(p23;q35) NPM-ALK chromosomal translocation. AB - The requirement for sufficient quantities of starting RNA has limited the ability to evaluate multiple transcripts using reverse transcriptase-polymerase chain reaction (RT-PCR). In this study, we demonstrate the utility of linear RNA amplification for RT-PCR analysis of multiple gene transcripts including a chromosomal translocation, using the t(2;5)(p23;q35) as a model. RNA from the t(2;5)-positive cell line, SU-DHL-1, and the t(2;5)-negative cell line, HUT-78, was extracted and exposed to two rounds of linear amplification. RT-PCR using cDNA from the resultant amplified (a) RNA and total RNA resulted in the 177 bp NPM-ALK fusion gene product from the SU-DHL-1 cell line, but not from aRNA or total RNA from the HUT-78 cell line. DNA sequencing of the RT-PCR products from total and aRNA of SU-DHL-1 cells demonstrated identical sequences corresponding to the NPM-ALK fusion gene. Evaluation of 25 snap-frozen tissue samples, including eight NPM-ALK-positive ALCLs demonstrated 100% concordance of t(2;5) detection between cDNA from total RNA and that from aRNA. Our results show that linear amplification of RNA can enhance starting RNA greater than 200-fold and can be used for rapid and specific detection of multiplex gene expression from a variety of sources. This method can generate a renewable archive of representative cDNA, which can be used for retrospective screening of stored samples as well as positive controls for the clinical molecular diagnostic laboratory. PMID- 14736823 TI - Comprehensive analysis of CBFbeta-MYH11 fusion transcripts in acute myeloid leukemia by RT-PCR analysis. AB - CBFbeta-MYH11 fusion transcripts are expressed in acute myeloid leukemias of the M4Eo subtype. Patients who express CBFbeta-MYH11 fusion transcripts respond favorably to high-dose chemotherapy and are generally spared allogeneic bone marrow transplantation. Hence it is important to identify this fusion in all patients with acute myeloid leukemia M4Eo leukemia. The fusion can be detected by cytogenetics, fluorescence in-situ hybridization (FISH), or by molecular analysis with RT-PCR. Multiple fusion transcripts arising as a result of various breakpoints in the CBFbeta and MYH11 have been identified. In this report we describe a comprehensive RT-PCR assay to identify all known fusion transcripts and provide an algorithm for molecular analysis of CBFbeta-MYH11 fusions from patient specimens. Further, identification of the fusion transcript by such an assay would help in the diagnosis and follow up of patients with cryptic inversion 16 translocations (such as patient 2 in this report) not detected by standard cytogenetics or FISH and for rational design of probes for quantitative analysis by real-time PCR. PMID- 14736824 TI - Classification of individual lung cancer cell lines based on DNA methylation markers: use of linear discriminant analysis and artificial neural networks. AB - The classification of small cell lung cancer (SCLC) and non-small cell lung cancer (NSCLC) can pose diagnostic problems due to inter-observer variability and other limitations of histopathology. There is an interest in developing classificatory models of lung neoplasms based on the analysis of multivariate molecular data with statistical methods and/or neural networks. DNA methylation levels at 20 loci were measured in 41 SCLC and 46 NSCLC cell lines with the quantitative real-time PCR method MethyLight. The data were analyzed with artificial neural networks (ANN) and linear discriminant analysis (LDA) to classify the cell lines into SCLC or into NSCLC. Models used either data from all 20 loci, or from five significant DNA methylation loci that were selected by a step-wise back-propagation procedure (PTGS2, CALCA, MTHFR, ESR1, and CDKN2A). The data were sorted randomly by cell line into 10 different data sets, each with training and testing subsets composed of 71 and 16 of the cases, respectively. Ten ANN models were trained using the 10 data sets: five using 20 variables, and five using the five variables selected by step-wise back-propagation. The ANN models with 20 input variables correctly classified 100% of the cell lines, while the models with only five variables correctly classified 87 to 100% of cases. For comparison, 10 different LDA models were trained and tested using the same data sets with either the original data or with logarithmically transformed data. Again, half of the models used all 20 variables while the others used only the five significant variables. LDA models provided correct classifications in 62.5% to 87.5% of cases. The classifications provided by all of the different models were compared with kappa statistics, yielding kappa values ranging from 0.25 to 1.0. We conclude that ANN models based on DNA methylation profiles can objectively classify SCLC and NSCLC cells lines with substantial to perfect concordance, while LDA models based on DNA methylation profiles provide poor to substantial concordance. Our work supports the promise of ANN analysis of DNA methylation data as a powerful approach for the development of automated methods for lung cancer classification. PMID- 14736825 TI - Quantification of parvovirus B19 DNA using COBAS AmpliPrep automated sample preparation and LightCycler real-time PCR. AB - The COBAS AmpliPrep instrument (Roche Diagnostics GmbH, D-68305 Mannheim, Germany) automates the entire sample preparation process of nucleic acid isolation from serum or plasma for polymerase chain reaction analysis. We report the analytical performance of the LightCycler Parvovirus B19 Quantification Kit (Roche Diagnostics) using nucleic acids isolated with the COBAS AmpliPrep instrument. Nucleic acids were extracted using the Total Nucleic Acid Isolation Kit (Roche Diagnostics) and amplified with the LightCycler Parvovirus B19 Quantification Kit. The kit combination processes 72 samples per 8-hour shift. The lower detection limit is 234 IU/ml at a 95% hit-rate, linear range approximately 10(4)-10(10) IU/ml, and overall precision 16 to 40%. Relative sensitivity and specificity in routine samples from pregnant women are 100% and 93%, respectively. Identification of a persistent parvovirus B19-infected individual by the polymerase chain reaction among 51 anti-parvovirus B19 IgM negative samples underlines the importance of additional nucleic acid testing in pregnancy and its superiority to serology in identifying the risk of parvovirus B19 transmission via blood or blood products. Combination of the Total Nucleic Acid Isolation Kit on the COBAS AmpliPrep instrument with the LightCycler Parvovirus B19 Quantification Kit provides a reliable and time-saving tool for sensitive and accurate detection of parvovirus B19 DNA. PMID- 14736826 TI - Real-time quantitative PCR of microdissected paraffin-embedded breast carcinoma: an alternative method for HER-2/neu analysis. AB - We studied the feasibility of using real-time quantitative PCR to determine HER-2 DNA amplification and mRNA expression in microdissected formalin-fixed, paraffin embedded breast tumors and compared this with standard immunohistochemistry (IHC) and fluorescent in situ hybridization (FISH) methods. Study cases (27 carcinomas and 3 ductal breast carcinoma in situ (DCIS) cases) showed varying Her-2 expression as determined by IHC (HercepTest). In carcinomas, there was a good correlation between HER-2 DNA amplification and strong HER-2 protein expression detected by FISH and IHC, respectively. A single DCIS case was amplified in FISH, but not in IHC. Both HER-2 gene amplification and expression could be quantified in microdissected paraffin-embedded tumors using real-time PCR, DNA and RNA being successfully detected in 146 of 150 (97%) and 141 of 150 (94%) samples, respectively. PCR analysis for HER-2 DNA amplification using the LightCycler HER2/neu DNA Quantification kit (Roche Molecular Biochemicals, Mannheim, Germany) correlated fairly well with IHC and FISH. All IHC HER-2 3+ tumors were amplified according to the kit, as was the FISH-amplified DCIS case. DNA-PCR identified five additional tumors as being amplified. Interestingly, all these scored 2+ with the HercepTest, but were negative using FISH. We believe that real-time quantitative PCR analysis of HER-2 DNA amplification following microdissection represents a useful supplementary or perhaps even an alternative technique for establishing HER-2 status in paraffin-embedded tumors. PMID- 14736827 TI - Molecular diagnosis of metastasizing oligodendroglioma: a case report. AB - We report the case of a suspicious parotid mass in which molecular determination of loss of heterozygosity (LOH) of chromosome arms 1p and 19q in combination with cytologic and immunohistochemical analysis defined the tumor to be metastatic oligodendroglioma. The patient was a 41-year-old woman who developed a World Health Organization grade II oligodendroglioma in her right frontal lobe at age 32, for which no adjuvant chemo- or radiotherapy was administered. Five years following this diagnosis, radiological assessment revealed a 10-centimeter mass in the tumor bed, suspicious for a recurrence. Resection of this lesion revealed an anaplastic oligodendroglioma (grade III) and adjuvant radiotherapy was given. Eleven months after this surgery the patient presented with a 5.5-cm subcutaneous, non-mobile, non-tender mass in the region of the right parotid gland. Fine needle aspiration (FNA) yielded highly cellular material, morphologically and immunohistochemically suspicious for oligodendroglioma. Molecular analysis of microsatellite loci residing on chromosome arms 1p and 19q was performed using DNA extracted from the patient's recurrent brain oligodendroglioma and the FNA specimen. This analysis revealed evidence of LOH at all eight of the microsatellite loci tested. The combination of cytologic and molecular findings defined the extracranial tumor to be metastatic oligodendroglioma. PMID- 14736829 TI - Expression of the tumor suppressor protein 14-3-3 sigma is down-regulated in invasive transitional cell carcinomas of the urinary bladder undergoing epithelial-to-mesenchymal transition. AB - The 14-3-3 proteins constitute a family of abundant, highly conserved and broadly expressed acidic polypeptides that are involved in the regulation of various cellular processes such as cell-cycle progression, cell growth, differentiation, and apoptosis. One member of this family, the 14-3-3 isoform sigma, is expressed only in epithelial cells and is frequently down-regulated in a variety of human cancers. To determine the prevalence of 14-3-3 sigma silencing in bladder cancer progression, we have studied the expression of this protein in normal urothelium and bladder transitional cell carcinomas (TCCs) of various grades and stages using two-dimensional gel electrophoresis in combination with Western blotting and immunohistochemistry. We show that the expression of 14-3-3 sigma is down regulated in invasive TCCs, particularly in lesions that are undergoing epithelial-to-mesenchymal conversion. Altered expression of 14-3-3 sigma in invasive TCCs is not due to increased externalization of the protein nor to an aberrant proliferative potential of neoplastic cells. Furthermore, we found that impaired 14-3-3 sigma expression is not associated with increased levels of the dominant-negative transcriptional regulator Delta Np63. Down-regulation of 14-3-3 sigma was confirmed by indirect immunofluorescence using a peptide-based rabbit polyclonal antibody specific for this protein. We also show that the expression of 14-3-3 sigma is highly up-regulated in pure squamous cell carcinomas. Taken together, these results provide evidence that deregulation of 14-3-3 sigma may play a key role in bladder cancer progression, in particular in differentiation events leading to epithelial-to-mesenchymal transition and stratified squamous metaplasia. PMID- 14736831 TI - PET/CT today and tomorrow. AB - Accurate anatomic localization of functional abnormalities seen with PET is known to be problematic. Even though nonspecific tracers such as 18F-FDG visualize certain normal anatomic structures, the spatial resolution is generally inadequate for localization of pathology. Combining PET with a high-resolution anatomic imaging modality such as CT can resolve the localization issue, as long as the images from the two modalities are accurately coregistered. However, software-based registration techniques have difficulty accounting for differences in patient positioning and involuntary movement of internal organs, often necessitating labor-intensive nonlinear mapping that may not converge to a satisfactory result. Acquiring both CT and PET images in the same scanner obviates the need for software registration and routinely provides accurately aligned images of anatomy and function in a single scan. DISCUSSION: A CT scanner positioned in tandem with a PET scanner and with a common patient couch and operating console has recently been explored as a solution to anatomic and functional image registration. Axial translation of the couch between the two modalities enables both CT and PET data to be acquired during a single imaging session. In addition, the CT images can be used to generate noiseless attenuation correction factors for the PET emission data. By minimizing patient movement between the CT and PET scans, and after accounting for the axial separation of the two modalities, accurately registered anatomic and functional images can be obtained. Since the introduction of the first PET/CT prototype a little over 5 years ago, several thousand cancer patients have been scanned on combined PET/CT devices. In the past 3 years, a number of commercial designs have become available featuring multidetector spiral CT scanners and high-performance PET devices. Initial experience has demonstrated an increased level of accuracy and confidence in the interpretation of the combined study compared with separate readings, particularly in the ability to distinguish pathology from normal physiologic uptake and to precisely localize abnormal foci. CONCLUSION: Combined PET/CT scanners represent an important evolution in technology that is helping to bring molecular imaging to the forefront in cancer diagnosis, staging, and therapy monitoring. PMID- 14736832 TI - PET/CT: panacea, redundancy, or something in between? AB - In the past decade, the integration of anatomic imaging and functional imaging has emerged as a new and promising diagnostic tool. Developments in software provided methods to integrate various modalities, such as PET, CT, MRI, and MR spectroscopy. The introduction of combined PET/CT scanners has boosted image fusion in this specific field and raised high expectations. Image fusion can be performed at 3 different levels: visual fusion, software fusion, and hardware fusion, each having strengths, weaknesses, and issues inherent to technique. Visual fusion is the traditional side-by-side reviewing of 2 separate modalities. Software image fusion provides evaluation of 2 modalities in 1 integrated image set. True hardware fusion of PET and CT does not exist at present. Currently, hardware fusion refers to a PET/CT scanner that consists of separate scanners, which positioned in line at a fixed distance, with projection of the PET image over the CT image. The suggested superiority of hardware fusion with these so called hybrid PET/CT scanners over software fusion has sparked debate. Because scientific data that unequivocally show that state-of-the-art software fusion is less accurate than hardware fusion (as provided in hybrid PET/CT scanners) are unavailable, the primacy of a combined PET/CT scanner over stand-alone PET and CT is more a matter of belief than of science. Further research comparing the overall performance of PET/CT scanners with that of separate scanners with software for image fusion is much needed. The continuous development of better software for image fusion and respiratory and cardiac gating is also needed, not only for PET and CT imaging but also for fusion of PET with MRI and CT with MRI. PMID- 14736833 TI - Acquisition protocol considerations for combined PET/CT imaging. AB - Since its introduction in 1998, dual-modality PET/CT imaging has received great attention in the medical community. For the first time, patients can be examined with both CT and PET in a single examination. A whole-body survey is the standard mode of acquisition. The CT images are used for anatomic reference of the tracer uptake patterns imaged in PET, as well as for attenuation correction of the PET data. The routine use of CT-based attenuation correction and user preferences for the quality and type of the CT examination have led to the introduction of different PET/CT scanning protocols. DISCUSSION: Two general approaches to PET/CT imaging can be distinguished today. One uses CT as a fast transmission source with little additional information for anatomic labeling. The other uses CT as a fast transmission source as well as a state-of-the-art diagnostic tool to maximize image quality using optimal acquisition parameters together with oral and intravenous contrast agents. Variations of these approaches share common concerns about image artifacts that result from mismatches in respiration and patient positioning between the CT and the PET examinations. Protocol requirements for the more complex radiologic PET/CT scenario also include alternative contrast application schemes or modifications to the attenuation correction procedure to handle CT contrast agents appropriately. CONCLUSION: High quality PET/CT studies can be provided routinely with existing PET/CT technology that is used efficiently by trained and motivated technologists and physicians. Only then will the potential diagnostic benefit of this new imaging modality be explored fully. PMID- 14736834 TI - Software approach to merging molecular with anatomic information. AB - Software image registration is a powerful and versatile tool that allows the fusion of molecular and anatomic information. Image registration can be applied to compare anatomic information with function, localize organs and lesions, and plan radiation therapy, biopsy, or surgery. Automatic volume-based image registration techniques have been devised for both linear and nonlinear image alignment. Challenges remain in the validation of the accuracy of software registration. Image registration has been applied clinically in neurology and oncology and may be particularly practical in radiotherapy applications. Potential new applications in cardiology could allow the combination of CT angiography with perfusion and viability images obtained by PET, SPECT, or MRI. Software methods allow versatility in the choice of modalities and facilitate retrospective and selective application. Fully automatic registration algorithms are needed for routine clinical applications. Connectivity, compatibility, and cooperation between various clinical departments are essential for the successful application of software-based image fusion in a hospital setting. PMID- 14736835 TI - PET/CT image navigation and communication. AB - The advent of multimodality imaging scanners combining PET and CT has led to a new paradigm in image display and presentation that raises new challenges in workstation interpretation software, image navigation, and communication. The essence of multimodality imaging is the ability to overlay imaging information from different modalities in a visually compelling fashion. This is accomplished by combining functional and anatomic data into multidimensional views using color encoding techniques that provide visual clues on the spatial distribution of image data. DISCUSSION: Combined PET/CT scanners provide spatially registered images from the two modalities acquired simultaneously in a single imaging session. Special reconstruction software and image display programs are required to rescale the native images from different spatial resolution into orthogonal or oblique reformatted planes in which data from PET images are color coded and superimposed on corresponding anatomic CT images. The color overlay technique allows the user to visually identify areas of high tracer activity and determine the underlying anatomic structure. Because of the multidimensional nature of the data, visualization requires interactive multidimensional navigation techniques that allow the viewer to move the visualization planes through three spatial directions and two additional dimensions. The fourth dimension is the continuum blend from PET to CT fusion, and the fifth is the dynamic range of the CT images that can be adjusted to display different tissue characteristics, such as bones, soft tissue, and lungs. Software tools currently available are often relatively complex, requiring the user to perform cumbersome maneuvers and time-consuming image manipulation to navigate through all dimensions and obtain adequate image settings and plane positioning for diagnostic interpretation of the image data. Moreover, the ability to convey these images to referring physicians is usually limited because of the lack of adequate viewing software. Distribution of results is usually performed instead through static "snapshots" of the fused images generated by the interpreting radiologist. The ability of the referring physician to navigate through the set of multimodality image data is thus limited. CONCLUSION: The wider adoption of multimodality PET/CT imaging techniques in routine clinical use will depend heavily on the development of more adequate image display and navigation tools that allow interpreting physicians to navigate easily and efficiently through multiple dimensions of data. Distribution of results to referring physicians and care providers also requires new tools for interactively reviewing the multimodality data, and current static images obtained from fused image data remain inadequate for proper visualization of the true content of images. PMID- 14736836 TI - To enhance or not to enhance? 18F-FDG and CT contrast agents in dual-modality 18F FDG PET/CT. AB - In combined PET/CT imaging, functional data provided by 18F-FDG increase diagnostic accuracy over conventional PET and CT in a variety of malignancies. The question to be raised is: how much is CT needed in PET/CT imaging? DISCUSSION: In stand-alone CT imaging, contrast agents are applied to aid differentiation of anatomic structures, improve lesion localization, and support lesion characterization. Based on our experience, the most important benefit when applying CT contrast agents in PET/CT imaging relates to more precise anatomic localization of pathology by differentiation of the lesion from its surrounding structures on CT. This benefit must be considered most profound in the head and neck as well as the abdomen and pelvis, where delineation of pathology from muscles, vascular structures, or the intestine is critical. CT contrast agents can be of additional value in tumors with only mild or no increase in 18F-FDG uptake by supporting lesion detection and characterization. The complementary role of CT contrast agents and molecular contrast for PET/CT imaging is addressed in this review. The authors provide specific indications as to which contrast enhanced CT data provide additional diagnostic value. CONCLUSION: Functional contrast aids lesion detection and characterization, whereas morphologic contrast improves lesion localization. CT contrast agents and PET tracers do not compete but rather are complementary in combined PET/CT imaging. PMID- 14736837 TI - Why most PET of lung and head-and-neck cancer will be PET/CT. AB - Experience in our and other institutions with PET/CT imaging of lung and head and neck cancers has shown that this new modality has higher specificity and sensitivity than PET alone and in certain settings even when compared to PET and CT viewed side by side. The largest experience exists with non-small cell lung cancer (NSCLC), in which it has been demonstrated that PET/CT is superior to PET and CT in T and in N staging. Superiority in M staging has yet to be demonstrated. CT contrast media enhancement is probably only necessary when a substantial mediastinal tumor component is present. In such cases, delineation of tumor from vascular structures is relevant. In ENT tumors, PET/CT also appears to be superior to PET, and probably also to PET and CT viewed side by side. Early information suggests that contrast media enhancement for staging may not be required, but the data available is still limited. In both settings, it is interesting to note that in a number of patients, second metachronous tumors are discovered with PET/CT, mainly localized in the GI tract. PMID- 14736838 TI - PET/CT in oncology: integration into clinical management of lymphoma, melanoma, and gastrointestinal malignancies. AB - PET/CT is a new imaging technology that has already found a number of clinical applications in oncologic imaging. Widespread introduction into clinical practice started approximately 2.5 years ago. Consequently, the available data are largely preliminary. Nevertheless, it can already be stated that the synthesis of structural and metabolic information improves the accuracy of primary staging and the detection of recurrent disease and has the realistic potential to change patient management in 10 to 20% of cases. PET/CT fusion images can directly guide biopsies or surgical interventions. This article summarizes preliminary data of PET/CT studies and highlights potential clinical applications for PET/CT, with particular emphasis on lymphoma, melanoma and gastrointestinal tumors. PMID- 14736839 TI - Why nearly all PET of abdominal and pelvic cancers will be performed as PET/CT. AB - Clinical experience at Johns Hopkins and published literature regarding PET/CT applications in the abdomen and pelvis are reviewed, and the strengths and limitations of this evolving technology are summarized. More than 2,700 whole body PET/CT scans including the abdomen and pelvis were performed for clinical indications by our nuclear medicine service from June 2001 through September 2003. Indications for these studies are reviewed, and our clinical impressions of diagnostic advantages and limitations of PET/CT are reported. Of the >2,700 whole body PET/CT scans performed at our institution, >90% were for known or suspected cancers. Primary abdominopelvic indications were second in frequency to thoracic indications. In addition, a comprehensive literature search was performed, and key articles related to PET/CT in the abdomen and pelvis were identified, reviewed, and summarized. Under the search term "PET/CT," 142 articles were identified under the National Library of Medicine Pub Med database, and a number of general findings are summarized. CONCLUSION: PET/CT allows for the accurate localization of foci of radiotracer uptake and their separation from normal structures. In our experience, the method is quantitatively accurate, rapid, and easily implemented, including contrast studies, in clinical practice in a wide range of abdominopelvic indications. Although artifacts can occur from a variety of causes, close attention to protocol details and patient immobilization reduces their frequency. Where systematically studied, PET/CT improves diagnostic accuracy compared with PET alone. It is anticipated that PET/CT will increasingly become the routine and preferred procedure for abdominopelvic evaluations with PET imaging. It has already become the preferred method at our center. PMID- 14736840 TI - Implementing biologic target volumes in radiation treatment planning for non small cell lung cancer. AB - 18F-FDG PET offers the radiation oncology community the ability to incorporate biologic information into radiation therapy targets. To date, most of the work in this arena has involved patients with non-small cell lung cancer (NSCLC). The literature suggests that biologic targeting with PET alters the radiation treatment volume significantly in 30%-60% of NSCLC patients for whom definitive therapy is planned. This is mostly the result of the incorporation of regional nodes with 18F-FDG avidity that were previously judged to be uninvolved by CT criteria. The development of the integrated PET/CT scanner is a valuable tool that improves diagnostic accuracy for staging this disease and will increase the accessibility of PET for radiation treatment planning. Its implementation into radiation treatment planning requires strong collaboration between radiation oncologists and nuclear physicians. In this report, we will review the literature on PET-based radiation treatment planning, its potential benefits, and future challenges. PMID- 14736842 TI - Reduced serotonin type 1A receptor binding in panic disorder. AB - Recent animal models suggest that disturbances in serotonin type-1A receptor (5 HT(1A)R) function may contribute to chronic anxiety, although it is not clear at all whether such models constitute relevant models for panic disorder (PD) in humans. The selective 5-HT(1A)R radioligand [18F]trans-4-fluoro-N-2-[4-(2 methoxyphenyl)piperazin-1-yl]ethyl]-N-(2-pyridyl)cyclohexanecarboxamide (FCWAY) permits in vivo assessment of central 5-HT(1A)R binding using positron emission tomography (PET). We studied 16 unmedicated symptomatic outpatients with PD and 15 matched healthy controls. Seven patients had an additional diagnosis of a current major depressive episode, however PD was the primary diagnosis. A 120 min PET study of 5-HT(1A)R binding was acquired using a GE Advance scanner in three dimensional mode. Using quantitative PET image analysis, regional values were obtained for [18F]-FCWAY volume of distribution (DV), corrected for plasma protein binding, and K1, the delivery rate of [18F]-FCWAY from plasma to tissue. MRI scanning was performed using a GE Signa Scanner (3.0 Tesla) to provide an anatomical framework for image analysis and partial volume correction of PET data. PD patients showed lower DV in the anterior cingulate (t = 4.3; p < 0.001), posterior cingulate (t = 4.1; p < 0.001), and raphe (t = 3.1; p = 0.004). Comparing patients with PD, patients with PD and comorbid depression, and healthy controls revealed that DVs did not differ between PD patients and PD patients with comorbid depression, whereas both patient groups differed significantly from controls. These results provide for the first time in vivo evidence for the involvement of 5-HT(1A)Rs in the pathophysiology of PD. PMID- 14736843 TI - M channels containing KCNQ2 subunits modulate norepinephrine, aspartate, and GABA release from hippocampal nerve terminals. AB - KCNQ subunits encode for the M current (I(KM)), a neuron-specific voltage dependent K+ current with a well established role in the control of neuronal excitability. In this study, by means of a combined biochemical, pharmacological, and electrophysiological approach, the role of presynaptic I(KM) in the release of previously taken up tritiated norepineprine (NE), GABA, and d-aspartate (d ASP) from hippocampal nerve terminals (synaptosomes) has been evaluated. Retigabine (RT) (0.01-30 microm), a specific activator of I(KM), inhibited [3H]NE, [3H]d-ASP, and [3H]GABA release evoked by 9 mm extracellular K+ ([K+]e). RT-induced inhibition of [3H]NE release was prevented by synaptosomal entrapment of polyclonal antibodies directed against KCNQ2 subunits, an effect that was abolished by antibody preabsorption with the KCNQ2 immunizing peptide; antibodies against KCNQ3 subunits were ineffective. Flupirtine (FP), a structural analog of RT, also inhibited 9 mm [K+]e-induced [3H]NE release, although its maximal inhibition was lower than that of RT. Electrophysiological studies in KCNQ2 transfected Chinese hamster ovary cells revealed that RT and FP (10 microm) caused a -19 and -9 mV hyperpolarizing shift, respectively, in the voltage dependence of activation of KCNQ2 K+ channels. In the same cells, the cognition enhancer 10,10-bis(4-pyridinylmethyl)-9(10H)-anthracenone (XE-991) (10 microm) blocked KCNQ2 channels and prevented their activation by RT (1-10 microm). Finally, both XE-991 (10-100 microm) and tetraethylammonium ions (100 microm) abolished the inhibitory effect of RT (1 microm) on [3H]NE release. These findings provide novel evidence for a major regulatory role of KCNQ2 K+ channel subunits in neurotransmitter release from rat hippocampal nerve endings. PMID- 14736844 TI - Locus ceruleus activation initiates delayed synaptic potentiation of perforant path input to the dentate gyrus in awake rats: a novel beta-adrenergic- and protein synthesis-dependent mammalian plasticity mechanism. AB - Norepinephrine, acting through beta-adrenergic receptors, is implicated in mammalian memory. In in vitro and in vivo studies, norepinephrine produces potentiation of the perforant path-dentate gyrus evoked potential; however, the duration and dynamics of norepinephrine-induced potentiation have not been explored over extended time periods. To characterize the long-term effects of norepinephrine on granule cell plasticity, the present study uses glutamatergic activation of the locus ceruleus (LC) to induce release of norepinephrine in the hippocampus of the awake rat and examines the subsequent modulation of the dentate gyrus evoked potential for 3 hr (short term) and 24 hr (long term) after LC activation. LC activation initiates a potentiation of the field EPSP slope observed 24 hr later. This late-phase potentiation of the synaptic potential is not preceded by early phase potentiation, although spike potentiation can be seen both immediately after, and 24 hr after, LC activation. Intracerebroventricular infusion of the beta-adrenergic antagonist, propranolol, or the protein synthesis inhibitor, anisomycin, before LC activation blocks the potentiation of perforant path input observed at 24 hr. The initiation of late-phase synaptic potentiation observed at 24 hr but not at the 3 hr after LC activation parallels the observation of a cAMP- and protein synthesis-dependent long-lasting synaptic facilitation in Aplysia that is not preceded by short-term synaptic facilitation. Locus ceruleus-initiated synaptic potentiation may selectively support long-term, rather than short-term, memory. The observation of selective initiation of long term synaptic facilitation in a mammalian brain, as in invertebrates, is additional evidence that these two forms of memory depend on separable biological mechanisms. PMID- 14736845 TI - The transition from development to motor control function in the corticospinal system. AB - During early postnatal development, corticospinal (CS) system stimulation, electrical or transcranial magnetic, is minimally effective in producing muscle contraction, despite having axon terminals that excite spinal neurons. Later, after stimulation becomes more effective, the cortical motor representation develops, and movements the system controls in maturity are expressed. We determined whether development of temporal facilitation (response enhancement produced by the second of a pair of pyramidal tract stimuli, or a higher stimulus multiple of a train of stimuli) correlated with these changes. Facilitation of the monosynaptic CS response was larger in older kittens and adults than younger kittens. When facilitation was strong, strong motor responses were evoked by pyramidal stimulation with small currents and few pulses. With strong facilitation in older kittens, corticospinal axon varicosities colocalize synaptophysin like adults, suggesting a presynaptic mechanism. With effective facilitation, control signals from the cortex can be sufficiently effective to provoke muscle contraction for guiding movements. PMID- 14736846 TI - The suprachiasmatic nucleus entrains, but does not sustain, circadian rhythmicity in the olfactory bulb. AB - The suprachiasmatic nucleus (SCN) of the hypothalamus has been termed the master circadian pacemaker of mammals. Recent discoveries of damped circadian oscillators in other tissues have led to the hypothesis that the SCN synchronizes and sustains daily rhythms in these tissues. We studied the effects of constant lighting (LL) and of SCN lesions on behavioral rhythmicity and Period 1 (Per1) gene activity in the SCN and olfactory bulb (OB). We found that LL had similar effects on cyclic locomotor and feeding behaviors and Per1 expression in the SCN but had no effect on rhythmic Period 1 expression in the OB. LL lengthened the period of locomotor and SCN rhythms by approximately 1.6 hr. After 2 weeks in LL, nearly 35% of rats lost behavioral rhythmicity. Of these, 90% showed no rhythm in Per1-driven expression in their SCN. Returning the animals to constant darkness rapidly restored their daily cycles of running wheel activity and gene expression in the SCN. In contrast, the OB remained rhythmic with no significant change in period, even when cultured from animals that had been behaviorally arrhythmic for 1 month. Similarly, we found that lesions of the SCN abolished circadian rhythms in behavior but not in the OB. Together, these results suggest that LL causes the SCN to lose circadian rhythmicity and its ability to coordinate daily locomotor and feeding rhythms. The SCN, however, is not required to sustain all rhythms because the OB continues to oscillate in vivo when the SCN is arrhythmic or ablated. PMID- 14736847 TI - Neurons and astrocytes respond to prion infection by inducing microglia recruitment. AB - The accumulation and activation of microglial cells at sites of amyloid prion deposits or plaques have been documented extensively. Here, we investigate the in vivo recruitment of microglial cells soon after intraocular injection of scrapie infected cell homogenate (hgtsc+) using immunohistochemistry on retinal sections. A population of CD11b/CD45-positive microglia was specifically detected within the ganglion and internal plexiform retinal cell layers by 2 d after intravitreal injection of hgtsc+. Whereas no chemotactism properties were ascribed to hgtsc+ alone, a massive migration of microglial cells was observed by incubating primary cultured neurons and astrocytes with hgtsc+ in a time- and concentration dependent manner. hgtsc+ triggered the recruitment of microglial cells by interacting with both neurons and astrocytes by upregulation of the expression levels of a broad spectrum of neuronal and glial chemokines. We show that, in vitro and in vivo, the microglia migration is at least partly under the control of chemokine receptor-5 (CCR-5) activation, because highly specific CCR-5 antagonist TAK-779 significantly reduced the migration rate of microglia. Activated microglia recruited in the vicinity of prion may, in turn, cause neuronal cell damage by inducing apoptosis. These findings provide insight into the understanding of the cell-cell communication that takes place during the development of prion diseases. PMID- 14736848 TI - Cortical synaptogenesis and motor map reorganization occur during late, but not early, phase of motor skill learning. AB - Extensive motor skill training induces reorganization of movement representations and synaptogenesis within adult motor cortex. Motor skill does not, however, develop uniformly across training sessions. It is characterized by an initial fast phase, followed by a later slow phase of learning. How cortical plasticity emerges during these phases is unknown. Here, we examine motor map topography and synapse number within rat motor cortex during the early and late phases of motor learning. Adult rats were placed in either a skilled or unskilled reaching condition (SRC and URC, respectively) for 3, 7, or 10 d. Intracortical microstimulation of layer V was used to determine the topography of forelimb movement representations within caudal forelimb area of motor cortex contralateral to the trained paw. Quantitative electron microscopy was used to measure the number of synapses per neuron within layer V. SRC animals showed significant increases in reaching accuracy after 3, 7, and 10 d of training. In comparison with URC animals, SRC animals had significantly larger distal forelimb representations after 10 d of training only. Furthermore, SRC animals had significantly more synapses per neuron than URC animals after 7 and 10 d of training. These results show that both motor map reorganization and synapse formation occur during the late phase of skill learning. Furthermore, synaptogenesis precedes map reorganization. We propose that motor map reorganization and synapse formation do not contribute to the initial acquisition of motor skills but represent the consolidation of motor skill that occurs during late stages of training. PMID- 14736849 TI - Columnar specificity of microvascular oxygenation and volume responses: implications for functional brain mapping. AB - Cortical neurons with similar properties are grouped in columnar structures and supplied by matching vascular networks. The hemodynamic response to neuronal activation, however, is not well described on a fine spatial scale. We investigated the spatiotemporal characteristics of microvascular responses to neuronal activation in rat barrel cortex using optical intrinsic signal imaging and spectroscopy. Imaging was performed at 570 nm to provide functional maps of cerebral blood volume (CBV) changes and at 610 nm to estimate oxygenation changes. To emphasize parenchymal rather than large vessel contributions to the functional hemodynamic responses, we developed an ANOVA-based statistical analysis technique. Perfusion-based maps were compared with underlying neuroanatomy with cytochrome oxidase staining. Statistically determined CBV responses localized accurately to individually stimulated barrel columns and could resolve neighboring columns with a resolution better than 400 microm. Both CBV and early oxygenation responses extended beyond anatomical boundaries of single columns, but this vascular point spread did not preclude spatial specificity. These results indicate that microvascular flow control structures providing targeted flow increases to metabolically active neuronal columns also produce finely localized changes in CBV. This spatial specificity, along with the high contrast/noise ratio, makes the CBV response an attractive mapping signal. We also found that functional oxygenation changes can achieve submillimeter specificity not only during the transient deoxygenation ("initial dip") but also during the early part of the hyperoxygenation. We, therefore, suggest that to optimize hemodynamic spatial specificity, appropriate response timing (using < or =2-3 sec changes) is more important than etiology (oxygenation or volume). PMID- 14736850 TI - Changes in the effect of spinal prostaglandin E2 during inflammation: prostaglandin E (EP1-EP4) receptors in spinal nociceptive processing of input from the normal or inflamed knee joint. AB - Inflammatory pain is caused by sensitization of peripheral and central nociceptive neurons. Prostaglandins substantially contribute to neuronal sensitization at both sites. Prostaglandin E2 (PGE2) applied to the spinal cord causes neuronal hyperexcitability similar to peripheral inflammation. Because PGE2 can act through EP1-EP4 receptors, we addressed the role of these receptors in the spinal cord on the development of spinal hyperexcitability. Recordings were made from nociceptive dorsal horn neurons with main input from the knee joint, and responses of the neurons to noxious and innocuous stimulation of the knee, ankle, and paw were studied after spinal application of recently developed specific EP1-EP4 receptor agonists. Under normal conditions, spinal application of agonists at EP1, EP2, and EP4 receptors induced spinal hyperexcitability similar to PGE2. Interestingly, the effect of spinal EP receptor activation changed during joint inflammation. When the knee joint had been inflamed 7-11 hr before the recordings, only activation of the EP1 receptor caused additional facilitation, whereas spinal application of EP2 and EP4 receptor agonists had no effect. Additionally, an EP3alpha receptor agonist reduced responses to mechanical stimulation. The latter also attenuated spinal hyperexcitability induced by spinal PGE2. In isolated DRG neurons, the EP3alpha agonist reduced the facilitatory effect of PGE2 on TTX-resistant sodium currents. Thus pronociceptive effects of spinal PGE2 can be limited, particularly under inflammatory conditions, through activation of an inhibitory splice variant of the EP3 receptor. The latter might be an interesting target for controlling spinal hyperexcitability in inflammatory pain states. PMID- 14736851 TI - Coordinate synaptic mechanisms contributing to olfactory cortical adaptation. AB - Anterior piriform cortex (aPCX) neurons rapidly filter repetitive odor stimuli despite relatively maintained input from mitral cells. This cortical adaptation is correlated with short-term depression of afferent synapses, in vivo. The purpose of this study was to elucidate mechanisms underlying this nonassociative neural plasticity using in vivo and in vitro preparations and to determine its role in cortical odor adaptation. Lateral olfactory tract (LOT)-evoked responses were recorded in rat aPCX coronal slices. Extracellular and intracellular potentials were recorded before and after simulated odor stimulation of the LOT. Results were compared with in vivo intracellular recordings from aPCX layer II/III neurons and field recordings in urethane-anesthetized rats stimulated with odorants. The onset, time course, and extent of LOT synaptic depression during both in vitro electrical and in vivo odorant stimulation methods were similar. Similar to the odor specificity of cortical odor adaptation in vivo, there was no evidence of heterosynaptic depression between independent inputs in vitro. In vitro evidence suggests at least two mechanisms contribute to this activity dependent synaptic depression: a rapidly recovering presynaptic depression during the initial 10-20 sec of the post-train recovery period and a longer lasting (approximately 120 sec) depression that can be blocked by the metabotropic glutamate receptor (mGluR) II/III antagonist (RS)-alpha-cyclopropyl-4 phosphonophenylglycine (CPPG) and by the beta-adrenergic receptor agonist isoproterenol. Importantly, in line with the in vitro findings, both adaptation of odor responses in the beta (15-35 Hz) spectral range and the associated synaptic depression can also be blocked by intracortical infusion of CPPG in vivo. PMID- 14736852 TI - Distance-dependent scaling of calcium transients evoked by backpropagating spikes and synaptic activity in dendrites of hippocampal interneurons. AB - Although interactions between backpropagating action potentials and synaptic stimulations have been extensively studied in pyramidal neurons, dendritic propagation and the summation of these signals in interneurons are not nearly as well known. In this study, two-photon imaging was used to explore the basic properties of dendritic calcium signaling in CA1 stratum radiatum interneurons. In contrast to hippocampal pyramidal neurons, the backpropagating action potential-evoked calcium transients in dendrites of interneurons underwent a distance-dependent increment. Although, in proximal dendrites, an increment could be attributed to a smaller dendrite diameter, distal dendrites did not show such dependence. Calcium responses in interneurons had a smaller amplitude, slower rise time, and decay than in pyramidal neurons. To explore the factors underlying the difference, we compared the calcium-binding capacity in interneurons and in pyramidal neurons. Our finding that endogenous calcium buffers had a higher level in interneurons may primarily explain the different kinetics and amplitudes of calcium transients. Synaptic stimulation-evoked calcium transients were also larger at distant dendritic locations. The spread of these signals was restricted to 12-13 microm long dendritic compartments. Supporting the reported lack of long term potentiation in these interneurons, we found only sublinear or linear summations of calcium responses to coincident synaptic inputs and backpropagating spikes. PMID- 14736853 TI - Role of matrix metalloproteinases in delayed neuronal damage after transient global cerebral ischemia. AB - Mechanisms of selective neuronal death in the hippocampus after global cerebral ischemia remain to be clarified. Here, we explored a possible role for matrix metalloproteinases (MMPs) in this phenomenon. Although many studies have demonstrated detrimental roles for the gelatinase MMP-9 in focal cerebral ischemia, how dysregulated MMP proteolysis influences global cerebral ischemia is less well understood. In this study, CD-1 mice were subjected to transient global ischemia. Transient occlusions of common carotid arteries for periods between 20 and 40 min led to increasing hippocampal neuronal death after 3 d. Gel zymography showed elevations in gelatinase (MMP-2 and MMP-9) activity. In situ zymography showed that gelatinase activity was mostly colocalized with neuron-specific nuclear protein-stained pyramidal neurons. Mice treated with the broad-spectrum metalloproteinase inhibitor BB-94 (50 mg/kg, i.p.) showed reduced hippocampal gelatinase activity after transient global cerebral ischemia and suffered significantly reduced hippocampal neuronal damage compared with vehicle-treated controls (p < 0.01). Additionally, hippocampal gelatinase activity and neuronal damage after transient global ischemia were also significantly reduced in MMP-9 knock-out mice compared with wild-type mice (p < 0.05). These data indicate a potential deleterious role for MMP-9 in the pathogenesis of delayed neuronal damage in the hippocampus after global cerebral ischemia. PMID- 14736854 TI - A C-terminal determinant of GluR6 kainate receptor trafficking. AB - Intracellular trafficking of ionotropic glutamate receptors is regulated predominantly by determinants in the cytoplasmic C-terminal domain of the subunit proteins. Although AMPA receptors are found at the vast majority of excitatory synapses, synaptic kainate receptors exhibit a much more restricted distribution, suggesting that specific mechanisms exist for selective trafficking of these receptor proteins. In this report, we define a critical forward trafficking motif that is necessary for surface expression of the glutamate receptor 6 (GluR6) kainate receptor as well as chimeric proteins containing only the GluR6 C terminal domain. The trafficking determinant was identified by tracking surface expression of green fluorescent protein-tagged GluR6 receptors with confocal immunofluorescence in COS-7 cells and cultured neurons and patch-clamp electrophysiology in human embryonic kidney 293 cells. Serial truncation and alanine site mutagenesis of the GluR6 subunit C terminus localized the critical motif to a seven amino acid stretch of predominantly basic residues. Alanine mutation of the trafficking motif reduced kainate receptor current amplitudes by >90% and resulted in retention of the mutated receptors in the endoplasmic reticulum. This forward trafficking domain is the first such identified for kainate receptors. PMID- 14736855 TI - Overexpression of adenosine kinase in epileptic hippocampus contributes to epileptogenesis. AB - Endogenous adenosine in the brain is thought to prevent the development and spread of seizures via a tonic anticonvulsant effect. Brain levels of adenosine are primarily regulated by the activity of adenosine kinase. To establish a link between adenosine kinase expression and seizure activity, we analyzed the expression of adenosine kinase in the brain of control mice and in a kainic acid induced mouse model of mesial temporal lobe epilepsy. Immunohistochemical analysis of brain sections of control mice revealed intense staining for adenosine kinase, mainly in astrocytes, which were more or less evenly distributed throughout the brain, as well as in some neurons, particularly in olfactory bulb, striatum, and brainstem. In contrast, hippocampi lesioned by a unilateral kainic acid injection displayed profound astrogliosis and therefore a significant increase in adenosine kinase immunoreactivity accompanied by a corresponding increase of enzyme activity, which paralleled chronic recurrent seizure activity in this brain region. Accordingly, seizures and interictal spikes were suppressed by the injection of a low dose of the adenosine kinase inhibitor 5-iodotubercidin. We conclude that overexpression of adenosine kinase in discrete parts of the epileptic hippocampus may contribute to the development and progression of seizure activity. PMID- 14736856 TI - Neural bases of set-shifting deficits in Parkinson's disease. AB - Patients with Parkinson's disease (PD) exhibit impairments in several cognitive functions similar to those observed in patients with prefrontal cortex (PFC) lesions. The physiological origins of these cognitive deficits are not well documented. Two mechanisms have been proposed: disruptions in corticostriatal circuits or a deficiency in frontal dopamine. We previously used functional magnetic resonance imaging (fMRI) in young healthy subjects to separate patterns of PFC and striatum activity during distinct phases of performance of the Wisconsin Card Sorting Task, a set-shifting task that reveals deficits in patients with PD. Here, the same fMRI protocol was used in PD patients and matched controls. Decreased activation was observed in the PD group compared with the matched control group in the ventrolateral PFC when receiving negative feedback and the posterior PFC when matching after negative feedback. In controls, these prefrontal regions specifically coactivated with the striatum during those stages of task performance. In contrast, greater activation was found in the PD group compared with the matched control group in prefrontal regions, such as the posterior and the dorsolateral PFC when receiving positive or negative feedback, that were not coactivated with the striatum in controls. These results suggest that both nigrostriatal dopamine depletion and intracortical dopamine deficiency may play a role in cognitive deficits in PD, depending on the involvement of the striatum in the task at hand. PMID- 14736857 TI - The development of mother-infant interactions after neonatal amygdala lesions in rhesus monkeys. AB - As part of ongoing studies on the neurobiology of socioemotional behavior in the nonhuman primate, we examined the development of mother-infant interactions in 24 macaque monkeys who received either bilateral amygdala or hippocampus ibotenic acid lesions, or a sham surgical procedure at 2 weeks of age. After surgery, the infants were returned to their mothers and reared with daily access to small social groups. Behavioral observations of the infants in dyads (mother-infant pairs alone), tetrads (two mother-infant pairs), and social groups (six mother infant pairs and one adult male) revealed species-typical mother-infant interactions for all lesion conditions, with the exception of increased physical contact time between the amygdala-lesioned infants and their mothers. Immediately after permanent separation from their mothers at 6 months of age, the infants were tested in a mother preference test that allowed the infants to choose between their mother and another familiar adult female. Unlike control and hippocampus-lesioned infants, the amygdala-lesioned infants did not preferentially seek proximity to their mother, nor did they produce distress vocalizations. Given the normal development of mother-infant interactions observed before weaning, we attribute the behavior of the amygdala-lesioned infants during the preference test to an impaired ability to perceive potential danger (i.e., separation from their mother in a novel environment), rather than to a disruption of the mother-infant relationship. These results are consistent with the view that the amygdala is not essential for fundamental aspects of social behavior but is necessary to evaluate potentially dangerous situations and to coordinate appropriate behavioral responses. PMID- 14736858 TI - Intracellular astrocyte calcium waves in situ increase the frequency of spontaneous AMPA receptor currents in CA1 pyramidal neurons. AB - Spontaneous neurotransmitter release and activation of group I metabotropic glutamate receptors (mGluRs) each play a role in the plasticity of neuronal synapses. Astrocytes may contribute to short- and long-term synaptic changes by signaling to neurons via these processes. Spontaneous whole-cell AMPA receptor (AMPAR) currents were recorded in CA1 pyramidal cells in situ while evoking Ca2+ increases in the adjacent stratum radiatum astrocytes by uncaging IP3. Whole-cell patch clamp was used to deliver caged IP3 and the Ca2+ indicator dye Oregon green BAPTA-1 to astrocytes. Neurons were patch-clamped and filled with Alexa 568 hydrazide dye to visualize their morphological relationship to the astrocyte. On uncaging of IP3, astrocyte Ca2+ responses reliably propagated as a wave into the very fine distal processes, synchronizing Ca2+ activity within astrocyte microdomains. The intracellular astrocyte Ca2+ wave coincided with a significant increase in the frequency of AMPA spontaneous EPSCs, but with no change in their kinetics. AMPAR current amplitudes were increased as well, but not significantly (p = 0.06). The increased frequency of AMPAR currents was sensitive to the group I mGluR antagonists LY367385 and 2-methyl-6-(phenylethynyl)-pyridine, suggesting that (1) astrocytes released glutamate in response to IP3 uncaging, and (2) glutamate released by astrocytes activated group I mGluRs to facilitate the release of glutamate from excitatory neuronal presynaptic boutons. These results extend previous studies, which have shown astrocyte modulation of neuronal activity in vitro and suggest that astrocyte-to-neuron signaling in intact tissue may contribute to synaptic plasticity. PMID- 14736859 TI - Firing patterns of type II spiral ganglion neurons in vitro. AB - Type I and type II spiral ganglion neurons convey auditory information from the sensory receptors in the cochlea to the CNS. The numerous type I neurons have been extensively characterized, but the small population of type II neurons with their unmyelinated axons are undetectable with most recording methods. Despite the paucity of information about the type II neurons, it is clear that they must have a significant role in sound processing because they innervate the large number of outer hair cells that are critical for maintaining normal responses to stimuli. To elucidate the function of type II neurons, we have developed an approach for studying their electrophysiological features in vitro. Type II neurons obtained from postnatal day 6-7 mice displayed distinctly different firing properties than type I neurons. They showed slower accommodation, lower action potential thresholds, and more prolonged responses to depolarizing current injection than the type I neurons. These differences were most evident in neurons from the basal, high-frequency region of the cochlea. The basal type I neurons displayed uniformly fast firing features, whereas the basal type II neurons showed particularly slow accommodation and responses to depolarization. Interestingly, neurons from the apical, low-frequency region of the cochlea showed the opposite trend. These data suggest that the type I and type II neurons have specialized electrophysiological characteristics tailored to their different roles in auditory signal processing. In particular, the type II neuron properties are consistent with cells in other sensory systems that receive convergent synaptic input for high-sensitivity stimulus detection. PMID- 14736860 TI - Heterogeneous requirement of NGF for sympathetic target innervation in vivo. AB - The neurotrophin nerve growth factor (NGF) plays a crucial role in the development of the sympathetic nervous system. In addition to being required for sympathetic neuron survival in vivo and in vitro, NGF has been shown to mediate axon growth in vitro. The role of NGF in sympathetic axon growth in vivo, however, is not clear because of its requirement for survival. This requirement can be circumvented by a concomitant deletion of Bax, a pro-apoptotic Bcl-2 family member, thus allowing an examination of the role of neurotrophins in axon growth independently of their function in cell survival. Here, we analyzed peripheral sympathetic target organ innervation in mice deficient for both NGF and Bax. In neonatal NGF-/-; Bax-/- mice, sympathetic target innervation was absent in certain organs (such as salivary glands), greatly reduced in others (such as heart), somewhat diminished in a few (such as stomach and kidneys), but not significantly different from control in some (such as trachea). At embryonic day 16.5, peripheral target sympathetic innervation was also reduced in NGF-/-; Bax-/- mice, with analogous variability for different organs. Interestingly, in some organs such as the spleen the precise location at which sympathetic axons become NGF-dependent for growth was evident. We thus show that NGF is required for complete peripheral innervation of both paravertebral and prevertebral sympathetic ganglia targets in vivo independently of its requirement for cell survival. Remarkably, target organs vary widely in their individual NGF requirements for sympathetic innervation. PMID- 14736861 TI - Posterior triangular thalamic neurons convey nociceptive messages to the secondary somatosensory and insular cortices in the rat. AB - This study investigated the responses of posterior triangular (PoT) thalamic neurons to tactile and noxious calibrated stimuli in anesthetized rats. We report here that 41% of PoT units responded to cutaneous stimulation, in most cases, by increasing strongly their firing. Forty-five percent of the responding units were nociceptive specific (NS), 19% were nociceptive nonspecific (NNS), and 36% were tactile. The NS units responded only to frankly noxious stimuli applied to relatively large receptive fields (several parts of the body). They encoded nociceptive temperatures chiefly in 46-50 degrees C ranges. The NNS units resembled NS units but also responded to innocuous stimuli. Tactile units responded chiefly to repeated innocuous stimuli applied to very small receptive fields (one to two fingers or vibrissae). A representative sample of PoT somatosensory neurons, characterized first by their response to innocuous and noxious cutaneous stimuli, were filled with juxtacellular injection of biotin dextran that made it possible to label adequately the soma, the dendrites, and the entire axon of PoT neurons. We observed that the axons of NS neurons terminated only in secondary somatosensory (S2) cortex, whereas the axons of NNS and tactile neurons projected chiefly to the insular cortex and the amygdala. In conclusion, our results demonstrate a spinal-PoT-S2/insular cortices nociceptive pathway that conveys nociceptive messages arising from lamina I and spinal neurons of deep laminas. Furthermore, our results demonstrate for the first time that projections of PoT neurons are correlated to their physiological properties. PMID- 14736862 TI - Menthol-induced Ca2+ release from presynaptic Ca2+ stores potentiates sensory synaptic transmission. AB - Menthol and many of its derivatives produce profound sensory and mental effects. The receptor for menthol has been cloned and named cold- and menthol-sensitive receptor-1 (CMR1) or transient receptor potential channel M8 (TRPM8) receptor. Using a dorsal root ganglion (DRG) and dorsal horn (DH) coculture system as a model for the first sensory synapse in the CNS, we studied menthol effects on sensory synaptic transmission and the underlying mechanisms. We found that menthol increased the frequency of miniature EPSCs (mEPSCs). The effects persisted under an extracellular Ca2+-free condition but were abolished by intracellular BAPTA and pretreatment with thapsigargin. Menthol-induced increases of mEPSC frequency were blocked by 2-aminoethoxydiphenylborane (2-APB) but not affected by the phospholipase C inhibitor U73122 [GenBank] or by the cADP receptor inhibitor 8-bromo-cADPR (8Br-cADPR). Double-patch recordings from DRG-DH pairs showed that menthol could potentiate evoked EPSCs (eEPSCs) and change the paired-pulse ratio of eEPSCs. A Ca2+ imaging study on DRG neurons demonstrated that menthol could directly release Ca2+ from intracellular Ca2+ stores. Menthol induced Ca2+ release was abolished by 2-APB but not affected by U73122 [GenBank] or 8Br-cADPR. Taken together, our results indicate that menthol can act directly on presynaptic Ca2+ stores of sensory neurons to release Ca2+, resulting in a facilitation of glutamate release and a modulation of neuronal transmission at sensory synapses. Expression of TRPM8 receptor on presynaptic Ca2+ stores, a novel localization for this ligand-gated ion channel, is also strongly suggested. PMID- 14736863 TI - Neuronal activity in the rodent dorsal striatum in sequential navigation: separation of spatial and reward responses on the multiple T task. AB - The striatum plays an important role in "habitual" learning and memory and has been hypothesized to implement a reinforcement-learning algorithm to select actions to perform given the current sensory input. Many experimental approaches to striatal activity have made use of temporally structured tasks, which imply that the striatal representation is temporal. To test this assumption, we recorded neurons in the dorsal striatum of rats running a sequential navigation task: the multiple T maze. Rats navigated a sequence of four T maze turns to receive food rewards delivered in two locations. The responses of neurons that fired phasically were examined. Task-responsive phasic neurons were active as rats ran on the maze (maze-responsive) or during reward receipt (reward responsive). Neither maze- nor reward-responsive neurons encoded simple motor commands: maze-responses were not well correlated with the shape of the rat's path and most reward-responsive neurons did not fire at similar rates at both food-delivery sites. Maze-responsive neurons were active at one or more locations on the maze, but these responses did not cluster at spatial landmarks such as turns. Across sessions the activity of maze-responsive neurons was highly correlated when rats ran the same maze. Maze-responses encoded the location of the rat on the maze and imply a spatial representation in the striatum in a task with prominent spatial demands. Maze-responsive and reward-responsive neurons were two separate populations, suggesting a divergence in striatal information processing of navigation and reward. PMID- 14736864 TI - Cutaneous and colonic rat DRG neurons differ with respect to both baseline and PGE2-induced changes in passive and active electrophysiological properties. AB - This study was designed to test the hypotheses that pain syndromes associated with specific body regions reflect unique properties of sensory neurons innervating these regions and/or unique responses of these afferents to tissue damage. Acutely dissociated adult rat dorsal root ganglia (DRG) neurons retrogradely labeled from either the colon or the glabrous skin of the hindpaw were studied by whole cell patch-clamp recording in current-clamp mode. Two populations of colonic afferent neurons were studied: pelvic afferents (arising from L(6), S(1), and S(2) DRG = LS DRG) and hypogastric/lumbar colonic afferents (arising from T(13), L(1), and L(2) DRG = TL DRG). Passive and active electrophysiological properties were studied before and after application prostaglandin E(2) (PGE(2)). We observed marked differences between cutaneous and colonic sensory neurons with respect to baseline passive and active electrophysiological properties as well as both the magnitude and pattern of PGE(2)-induced changes in excitability, passive, and active properties. There were also significant differences between TL and LS neurons with respect to baseline and PGE(2)-induced changes in several passive and active electrophysiological properties. Our results suggest that differences between cutaneous and colonic neurons reflect differences in pattern and/or density of ionic currents present in the plasma membrane. More interestingly, the ionic currents underlying the PGE(2)-induced sensitization of cutaneous neurons appeared to differ from those underlying the sensitization of colonic neurons. The implication of this observation is that it may be possible, in fact necessary, to treat pain arising from specific body regions with unique therapeutic interventions. PMID- 14736865 TI - Stochastic resonance in muscle receptors. AB - Noise is generally considered to have deleterious effects on the sensitivity of a signal detection system. There are, however, several mechanisms whereby the addition of noise to the input of a system can in fact improve sensitivity. One such mechanism is stochastic resonance. Although first proposed in 1981, conclusive experimental evidence for "fully tuneable stochastic resonance" in biological systems has not previously been reported. Evidence of fully tuneable stochastic resonance in the response of afferents from Golgi tendon organs and the primary and secondary endings of muscle spindles to imposed muscle length changes is presented. PMID- 14736866 TI - Direct evidence that toll-like receptor 9 (TLR9) functionally binds plasmid DNA by specific cytosine-phosphate-guanine motif recognition. AB - Cytosine-phosphate-guanine (CpG) motifs in bacterial DNA are known to activate the mammalian immune system, and this activation is thought to depend on the Toll like receptor 9 (TLR9) signaling pathway. Previous studies strongly suggested that TLR9 is involved as the specific receptor for CpG motifs but did not provide direct evidence of their interaction. In this study, we demonstrate for the first time that murine TLR9 binds an unmethylated CpG-containing plasmid. This interaction is sequence-specific and is influenced by the methylation status of the plasmid. Furthermore, we demonstrate that this interaction leads to the activation of the NF-kappaB pathway in mTLR9-expressing cells. Our results provide a molecular basis for the interaction between CpG-DNA and TLR9. PMID- 14736867 TI - Serine 171, a conserved residue in the gamma-aminobutyric acid type A (GABAA) receptor gamma2 subunit, mediates subunit interaction and cell surface localization. AB - Serine 171 in the GABA(A) receptor gamma2 subunit is highly conserved in the ligand-gated ion channel superfamily. In this paper, we report that mutating serine 171 within gamma2 to glycine or cysteine prevents the interaction of gamma2 with alpha2 and beta1 when these subunits are co-expressed in human embryo kidney 293 cells, resulting in intracellular retention of gamma2. Structure analysis based on a three-dimensional homology model of gamma2 (Ernst, M., Brauchart, D., Boresch, S., and Sieghart, W. (2003) Neuroscience 119, 933-943) reveals that serine 171 may play a critical role in the formation and stabilization of an exposed turn structure that is part of the subunit interaction site. Mutation of serine 171 in the gamma2 subunit could therefore result in alteration of the structure of the subunit interaction site, preventing correct subunit assembly. PMID- 14736869 TI - Hydrogen bonds involved in binding the Qi-site semiquinone in the bc1 complex, identified through deuterium exchange using pulsed EPR. AB - Exchangeable protons in the immediate neighborhood of the semiquinone (SQ) at the Qi-site of the bc1 complex (ubihydroquinone:cytochrome c oxidoreductase (EC 1.10.2.2)) from Rhodobacter sphaeroides have been characterized using electron spin echo envelope modulation (ESEEM) and hyperfine sublevel correlation spectroscopy (HYSCORE) and visualized by substitution of H2O by 2H2O. Three exchangeable protons interact with the electron spin of the SQ. They possess different isotropic and anisotropic hyperfine couplings that allow a clear distinction between them. The strength of interactions indicates that the protons are involved in hydrogen bonds with SQ. The hyperfine couplings differ from values typical for in-plane hydrogen bonds previously observed in model experiments. It is suggested that the two stronger couplings involve formation of hydrogen bonds with carbonyl oxygens, which have a significant out-of-plane character due to the combined influence of bulky substituents and the protein environment. These two hydrogen bonds are most probably to side chains suggested from crystallographic structures (His-217 and Asp-252 in R. sphaeroides). Assignment of the third hydrogen bond is more ambiguous but may involve either a bond between Asn-221 and a methoxy O-atom or a bond to water. The structural and catalytic roles of the exchangeable protons are discussed in the context of three high resolution crystallographic structures for mitochondrial bc1 complexes. Potential H-bonds, including those to water molecules, form a network connecting the quinone (ubiquinone) occupant and its ligands to the propionates of heme bH and the external aqueous phase. They provide pathways for exchange of protons within the site and with the exteriors, needed to accommodate the different hydrogen bonding requirements of different quinone species during catalysis. PMID- 14736868 TI - Capping protein binding to S100B: implications for the tentacle model for capping the actin filament barbed end. AB - S100B binds tightly to a 12-amino acid peptide derived from heterodimeric capping protein. In native intact capping protein, this sequence is in the C terminus of the alpha-subunit, which is important for capping the actin filament. This C terminal region is proposed to act as a flexible "tentacle," extending away from the body of capping protein in order to bind actin. To this hypothesis, we analyzed the interaction between S100B and capping protein in solution. The C terminal 28 amino acids of the alpha-subunit, the proposed tentacle, bound to S100B as a free synthetic peptide or a glutathione S-transferase fusion (K(d) approximately 0.4-1 microm). In contrast, S100B did not bind to whole native capping protein or functionally affect its capping activity. S100B does not bind, with any significant affinity, to the proposed alpha-tentacle sequence of whole native capping protein in solution. In the NMR structure of S100B complexed with the alpha-subunit-derived 12-amino acid peptide, the hydrophobic side of a short alpha-helix in the peptide, containing an important tryptophan residue, contacts S100B. In the x-ray structure of native capping protein, the corresponding sequence of the alpha-subunit C terminus, including Trp(271), interacts closely with the body of the protein. Therefore, our results suggest the alpha-subunit C terminus is not mobile as predicted by the tentacle model. Addition of non-ionic detergent allowed whole capping protein to bind weakly to S100B, indicating that the alpha-subunit C terminus can be mobilized from the surface of the capping protein molecule, presumably by weakening the hydrophobic binding at the contact site. PMID- 14736870 TI - Evidence of an intramolecular interaction between the two domains of the BlaR1 penicillin receptor during the signal transduction. AB - The BlaR1 protein is a penicillin-sensory transducer involved in the induction of the Bacillus licheniformis beta-lactamase. The amino-terminal domain of the protein exhibits four transmembrane segments (TM1-TM4) that form a four-alpha helix bundle embedded in the plasma bilayer. The carboxyl-terminal domain of 250 amino acids (BlaR-CTD) fused at the carboxyl end of TM4 possesses the amino acid sequence signature of penicillin-binding proteins. This membrane topology suggests that BlaR-CTD and the BlaR-amino-terminal domain are responsible for signal reception and signal transduction, respectively. With the use of phage display experiments, we highlight herein an interaction between BlaR-CTD and the extracellular, 63-amino acid L2 loop connecting TM2 and TM3. This interaction does not occur in the presence of penicillin. This result suggests that binding of the antibiotic to BlaR1 might entail the release of the interaction between L2 and BlaR-CTD, causing a motion of the alpha-helix bundle and transfer of the information to the cytoplasm of the cell. In addition, fluorescence spectroscopy, CD, and Fourier transform IR spectroscopy experiments indicate that in contrast to the behavior of the corresponding Staphylococcus aureus protein, the beta lactam antibiotic does not induce a drastic conformational change in B. licheniformis BlaR-CTD. PMID- 14736871 TI - Molecular determinants involved in the allosteric control of agonist affinity in the GABAB receptor by the GABAB2 subunit. AB - The gamma-aminobutyric acid type B (GABAB) receptor is an allosteric complex made of two subunits, GABAB1 (GB1) and GABAB2 (GB2). Both subunits are composed of an extracellular Venus flytrap domain (VFT) and a heptahelical domain (HD). GB1 binds GABA, and GB2 plays a major role in G-protein activation as well as in the high agonist affinity state of GB1. How agonist affinity in GB1 is regulated in the receptor remains unknown. Here, we demonstrate that GB2 VFT is a major molecular determinant involved in this control. We show that isolated versions of GB1 and GB2 VFTs in the absence of the HD and C-terminal tail can form hetero oligomers as shown by time-resolved fluorescence resonance energy transfer (based on HTRF technology). GB2 VFT and its association with GB1 VFT controlled agonist affinity in GB1 in two ways. First, GB2 VFT exerted a direct action on GB1 VFT, as it slightly increased agonist affinity in isolated GB1 VFT. Second and most importantly, GB2 VFT prevented inhibitory interaction between the two main domains (VFT and HD) of GB1. According to this model, we propose that GB1 HD prevents the possible natural closure of GB1 VFT. In contrast, GB2 VFT facilitates this closure. Finally, such inhibitory contacts between HD and VFT in GB1 could be similar to those important to maintain the inactive state of the receptor. PMID- 14736872 TI - The crystal structure of Synechocystis hemoglobin with a covalent heme linkage. AB - The x-ray crystal structure of Synechocystis hemoglobin has been solved to a resolution of 1.8 A. The conformation of this structure is surprisingly different from that of the previously reported solution structure, probably due in part to a covalent linkage between the heme 2-vinyl and His117 that is present in the crystal structure but not in the structure solved by NMR. Synechocystis hemoglobin is a hexacoordinate hemoglobin in which the heme iron is coordinated by both the proximal and distal histidines. It is also a member of the "truncated hemoglobin" family that is much shorter in primary structure than vertebrate and plant hemoglobins. In contrast to other truncated hemoglobins, the crystal structure of Synechocystis hemoglobin displays no "ligand tunnel" and shows that several important amino acid side chains extrude into the solvent instead of residing inside the heme pocket. The stereochemistry of hexacoordination is compared with other hexacoordinate hemoglobins and cytochromes in an effort to illuminate factors contributing to ligand affinity in hexacoordinate hemoglobins. PMID- 14736873 TI - Characterization of four autonomous repression domains in the corepressor receptor interacting protein 140. AB - Receptor interacting protein (RIP) 140 is a corepressor that can be recruited to nuclear receptors by means of LXXLL motifs. We have characterized four distinct autonomous repression domains in RIP140, termed RD1-4, that are highly conserved in mammals and birds. RD1 at the N terminus represses transcription in the presence of trichostatin A, suggesting that it functions by a histone deacetylase (HDAC)-independent mechanism. The repressive activity of RD2 is dependent upon carboxyl-terminal binding protein recruitment to two specific binding sites. Use of specific inhibitors indicates that RD2, RD3, and RD4 are capable of functioning by HDAC-dependent and HDAC-independent mechanisms, depending upon cell type. PMID- 14736874 TI - Cell surface expression of alpha1D-adrenergic receptors is controlled by heterodimerization with alpha1B-adrenergic receptors. AB - alpha(1)-Adrenergic receptors (ARs) belong to the large Class I G protein-coupled receptor superfamily and comprise three subtypes (alpha(1A), alpha(1B), and alpha(1D)). Previous work with heterologously expressed C-terminal green fluorescent protein (GFP)-tagged alpha(1)-ARs showed that alpha(1A)- and alpha(1B)-ARs localize to the plasma membrane, whereas alpha(1D)-ARs accumulate intracellularly. We recently showed that alpha(1D)- and alpha(1B)-ARs form heterodimers, whereas alpha(1D)- and alpha(1A)-ARs do not. Here, we examined the role of heterodimerization in regulating alpha(1D)-AR localization using both confocal imaging of GFP- or CFP-tagged alpha(1)-ARs and a luminometer-based surface expression assay in HEK293 cells. Co-expression with alpha(1B)-ARs caused alpha(1D)-ARs to quantitatively translocate to the cell surface, but co expression with alpha(1A)-ARs did not. Truncation of the alpha(1B)-AR extracellular N terminus or intracellular C terminus had no effect on surface expression of alpha(1D)-ARs, suggesting primary involvement of the hydrophobic core. Co-transfection with an uncoupled mutant alpha(1B)-AR (Delta12alpha(1B)) increased both alpha(1D)-AR surface expression and coupling to norepinephrine stimulated Ca(2+) mobilization. Finally, GFP-tagged alpha(1D)-ARs were not detected on the cell surface when expressed in rat aortic smooth muscle cells that express no endogenous ARs, but were almost exclusively localized on the surface when expressed in DDT(1)MF-2 cells, which express endogenous alpha(1B) ARs. These studies demonstrate that alpha(1B)/alpha(1D)-AR heterodimerization controls surface expression and functional coupling of alpha(1D)-ARs, the N- and C-terminal domains are not involved in this interaction, and that alpha(1B)-AR G protein coupling is not required. These observations may be relevant to many other Class I G protein-coupled receptors, where the functional consequences of heterodimerization are still poorly understood. PMID- 14736875 TI - TIA proteins are necessary but not sufficient for the tissue-specific splicing of the myosin phosphatase targeting subunit 1. AB - We are using the tissue-specific splicing of myosin phosphatase targeting subunit (MYPT1) as a model to investigate smooth muscle phenotypic diversity. We previously identified a U-rich intronic enhancer flanking the 5' splice site (IE1), and a bipartite exonic enhancer/suppressor, that regulate splicing of the MYPT1 central alternative exon. Here we show that T-cell inhibitor of apoptosis (TIA-1) and T-cell inhibitor of apoptosis-related (TIAR) proteins bind to the IE1. Co-transfection of TIA expression vectors with a MYPT1 mini-gene construct increase splicing of the central alternative exon. TIA proteins do not enhance splicing when the palindromic exonic splicing enhancer (ESE) is mutated, indicating that TIAs are necessary but not sufficient for splicing. The ESE specifically binds SRp55 and SRp20 proteins, supporting a model in which both SR and TIA proteins binding to their cis-elements are required for the recruitment of the splicing complex to a weak 5' splice site. Inactivation of TIA proteins in the DT40 cell line (TIA-1(-/-)TIAR(+/-)) reduced the splicing of the central alternative exon of the endogenous MYPT1 as well as stably transfected MYPT1 minigene constructs. Splicing of the MYPT1 3' alternative exon and the MLC(17) alternative exon were unaffected, suggesting that TIA proteins regulate a subset of smooth muscle/nonmuscle alternative splicing reactions. Finally, reduced RNA binding and reduced expression of the TIA and SR proteins in phasic (gizzard) smooth muscle around hatching coincided with the switch from exon inclusion to exon skipping, suggesting that loss of TIA and SR enhancer activity may play a role in the developmental switch in MYPT1 splicing. PMID- 14736876 TI - A WT1 co-regulator controls podocyte phenotype by shuttling between adhesion structures and nucleus. AB - Glomerular podocyte differentiation state is critical for filtration barrier function and is regulated by WT1, a zinc finger transcription factor. A yeast two hybrid assay identified a novel, WT1-interacting protein (WTIP) that maps to human chromosome 19q13.1, a region with genes linked to familial focal segmental glomerulosclerosis. The domain structure of WTIP is similar to the zyxin subfamily of cytosolic LIM domain-containing proteins, which contain three carboxyl-terminal LIM protein-protein interaction domains and a proline-rich, pre LIM region with a nuclear export signal. Other LIM domain-containing proteins (zyxin and mouse muscle LIM protein) did not interact with WT1 in two-hybrid assays, and WTIP did not interact with an unrelated transcription factor, LMX1B. WTIP mRNA was detected in cultured podocytes and was developmentally regulated, with expression peaking in mouse kidney at embryonic day 15-16 (E15-E16) in kidney but persisting into adulthood. In situ hybridization demonstrated WTIP expression in developing E15 glomeruli and in cultured podocytes. The partial WTIP clone, which interacted with WTIP in the two-hybrid assay, co-localized with WT1 in nuclei, co-precipitated with WT1, and inhibited WT1-dependent transcriptional activation of the amphiregulin promoter. In contrast, full-length WTIP was excluded from cell nuclei, but after the addition of leptomycin B, an inhibitor of Crm1-mediated nuclear export, it accumulated in the nucleus and co precipitated with WT1 in whole cell lysates. Epitope-tagged WTIP co-localized with the adaptor protein CD2AP (CMS) in podocyte actin spots and with Mena at cell-cell junctions. We propose that WTIP monitors slit diaphragm protein assembly as part of a multiple protein complex, linking this specialized adhesion junction to the actin cytoskeleton, and shuttles into the nucleus after podocyte injury, providing a mechanism whereby changes in slit diaphragm structure modulate gene expression. PMID- 14736877 TI - An R111C polymorphism in wild turkey cardiac troponin I accompanying the dilated cardiomyopathy-related abnormal splicing variant of cardiac troponin T with potentially compensatory effects. AB - Cardiac muscle contraction is regulated by Ca(2+) through the troponin complex consisting of three subunits: troponin C (TnC), troponin T (TnT), and troponin I (TnI). We reported previously that the abnormal splicing of cardiac TnT in turkeys with dilated cardiomyopathy resulted in a greater binding affinity to TnI. In the present study, we characterized a polymorphism of cardiac TnI in the heart of wild turkeys. cDNA cloning and sequencing of the novel turkey cardiac TnI revealed a single amino acid substitution, R111C. Arg(111) in avian cardiac TnI corresponds to a Lys in mammals. This residue is conserved in cardiac and skeletal muscle TnIs across the vertebrate phylum, implying a functional importance. In the partial crystal structure of cardiac troponin, this amino acid resides in an alpha-helix that directly contacts with TnT. Structural modeling indicates that the substitution of Cys for Arg or Lys at this position would not disrupt the global structure of troponin. To evaluate the functional significance of the different size and charge between the Arg and Cys side chains, protein binding assays using purified turkey cardiac TnI expressed in Escherichia coli were performed. The results show that the R111C substitution lowered binding affinity to TnT, which is potentially compensatory to the increased TnI-binding affinity of the cardiomyopathy-related cardiac TnT splicing variant. Therefore, the fixation of the cardiac TnI Cys(111) allele in the wild turkey population and the corresponding functional effect reflect an increased fitness value, suggesting a novel target for the treatment of TnT myopathies. PMID- 14736878 TI - Macrophage migration inhibitory factor is induced by thrombin and factor Xa in endothelial cells. AB - Macrophage migration inhibitory factor (MIF), a proinflammatory cytokine, has been shown to play a role in wound-healing processes. In this study, we investigated whether protease-activated receptor (PAR)-1 and PAR-2 mediated MIF expression in human endothelial cells. Thrombin, factor Xa (FXa), and trypsin induced MIF expression in human dermal microvascular endothelial cells and human umbilical vein endothelial cells, but other proteases, including kallikrein and urokinase, failed to do so. Thrombin-induced MIF mRNA expression was significantly reduced by the thrombin-specific inhibitor hirudin. Thrombin receptor activation peptide-6, a synthetic PAR-1 peptide, induced MIF mRNA expression, suggesting that PAR-1 mediates MIF expression in response to thrombin. The effects of FXa were blocked by antithrombin III, but not by hirudin, indicating that FXa might enhance MIF production directly rather than via thrombin stimulation. The synthetic PAR-2 peptide SLIGRL-NH(2) induced MIF mRNA expression, showing that PAR-2 mediated MIF expression in response to FXa. Concerning the signal transduction, a mitogen-activated protein kinase kinase inhibitor (PD98089) and a nuclear factor (NF)-kappaB inhibitor (SN50) suppressed the up-regulation of MIF mRNA in response to thrombin, FXa, and PAR-2 agonist stimulation, whereas a p38 inhibitor (SB203580) had little effect. These facts indicate that up-regulation of MIF by thrombin or FXa is regulated by p44/p42 mitogen-activated protein kinase-dependent pathways and NF-kappaB-dependent pathways. Moreover, we found that PAR-1 and PAR-2 mRNA expression in endothelial cells was enhanced by MIF. Furthermore, we examined the inflammatory response induced by PAR-1 and PAR-2 agonists injected into the mouse footpad. As shown by footpad thickness, an indicator of inflammation, MIF-deficient mice (C57BL/6) were much less sensitive to either PAR-1 or PAR-2 agonists than wild-type mice. Taken together, these results suggest that MIF contributes to the inflammatory phase of the wound healing process in concert with thrombin and FXa via PAR-1 and PAR-2. PMID- 14736879 TI - Microarray analysis supports a role for ccaat/enhancer-binding protein-beta in brain injury. AB - CCAAT/enhancer-binding protein-beta (C/EBPbeta) is a transcription factor that plays an important role in regulating cell growth and differentiation. This protein plays a central role in lymphocyte and adipocyte differentiation and hepatic regeneration and in the control of inflammation and immunity in the liver and in cells of the myelomonocytic lineage. Our previous studies suggested that this protein could also have important functions in the brain. Therefore, we were interested in the identification of downstream targets of this transcription factor in cells of neural origin. We performed cDNA microarray analysis and found that a total of 48 genes were up-regulated in C/EBPbeta-overexpressing neuronal cells. Of the genes that displayed significant changes in expression, several were involved in inflammatory processes and brain injury. Northern blot analysis confirmed the up-regulation of ornithine decarboxylase, 24p3/LCN2, GRO1/KC, spermidine/spermine N(1)-acetyltransferase, xanthine dehydrogenase, histidine decarboxylase, decorin, and TM4SF1/L6. Using promoter-luciferase reporter transfection assays, we showed the ornithine decarboxylase and 24p3 genes to be biological downstream targets of C/EBPbeta in neuroblastoma cells. Moreover, the levels of C/EBPbeta protein were significantly induced after neuronal injury, which was accompanied by increased levels of cyclooxygenase-2 enzyme. This strongly supports the concept that C/EBPbeta may play an important role in brain injury. PMID- 14736880 TI - Flexible N-terminal region of prion protein influences conformation of protease resistant prion protein isoforms associated with cross-species scrapie infection in vivo and in vitro. AB - Transmissible spongiform encephalopathy (TSE) diseases are characterized by the accumulation in brain of an abnormal protease-resistant form of the host-encoded prion protein (PrP), PrP-res. PrP-res conformation differs among TSE agents derived from various sources, and these conformational differences are thought to influence the biological characteristics of these agents. In this study, we introduced deletions into the flexible N-terminal region of PrP (residues 34-124) and investigated the effect of this region on the conformation of PrP-res generated in an in vitro cell-free conversion assay. PrP deleted from residues 34 to 99 generated 12-16-kDa protease-resistant bands with intact C termini but variable N termini. The variable N termini were the result of exposure of new protease cleavage sites in PrP-res between residues 130 and 157, suggesting that these new cleavage sites were caused by alterations in the conformation of the PrP-res generated. Similarly truncated 12-16-kDa PrP bands were also identified in brain homogenates from mice infected with mouse-passaged hamster scrapie as well as in the cell-free conversion assay using conditions that mimicked the hamster/mouse species barrier to infection. Thus, by its effects on PrP-res conformation, the flexible N-terminal region of PrP seemed to influence TSE pathogenesis and cross-species TSE transmission. PMID- 14736881 TI - Ca(2+) signaling by TRPC3 involves Na(+) entry and local coupling to the Na(+)/Ca(2+) exchanger. AB - TRPC3 has been suggested as a key component of phospholipase C-dependent Ca(2+) signaling. Here we investigated the role of TRPC3-mediated Na(+) entry as a determinant of plasmalemmal Na(+)/Ca(2+) exchange. Ca(2+) signals generated by TRPC3 overexpression in HEK293 cells were found to be dependent on extracellular Na(+), in that carbachol-stimulated Ca(2+) entry into TRPC3 expressing cells was significantly suppressed when extracellular Na(+) was reduced to 5 mm. Moreover, KB-R9743 (5 microm) an inhibitor of the Na(+)/Ca(2+) exchanger (NCX) strongly suppressed TRPC3-mediated Ca(2+) entry but not TRPC3-mediated Na(+) currents. NCX1 immunoreactivity was detectable in HEK293 as well as in TRPC3-overexpressing HEK293 cells, and reduction of extracellular Na(+) after Na(+) loading with monensin resulted in significant rises in intracellular free Ca(2+) (Ca(2+)(i)) of HEK293 cells. Similar rises in Ca(2+)(i) were recorded in TRPC3-overexpressing cells upon the reduction of extracellular Na(+) subsequent to stimulation with carbachol. These increases in Ca(2+)(i) were associated with outward membrane currents at positive potentials and inhibited by KB-R7943 (5 microm), chelation of extracellular Ca(2+), or dominant negative suppression of TRPC3 channel function. This suggests that Ca(2+) entry into TRPC3-expressing cells involves reversed mode Na(+)/Ca(2+) exchange. Cell fractionation experiments demonstrated co-localization of TRPC3 and NCX1 in low density membrane fractions, and co immunoprecipitation experiments provided evidence for association of TRPC3 and NCX1. Glutathione S-transferase pull-down experiments revealed that NCX1 interacts with the cytosolic C terminus of TRPC3. We suggest functional and physical interaction of nonselective TRPC cation channels with NCX proteins as a novel principle of TRPC-mediated Ca(2+) signaling. PMID- 14736882 TI - The functional role of the binuclear metal center in D-aminoacylase: one-metal activation and second-metal attenuation. AB - Our structural comparison of the TIM barrel metal-dependent hydrolase(-like) superfamily suggests a classification of their divergent active sites into four types: alphabeta-binuclear, alpha-mononuclear, beta-mononuclear, and metal independent subsets. The d-aminoacylase from Alcaligenes faecalis DA1 belongs to the beta-mononuclear subset due to the fact that the catalytically essential Zn(2+) is tightly bound at the beta site with coordination by Cys(96), His(220), and His(250), even though it possesses a binuclear active site with a weak alpha binding site. Additional Zn(2+), Cd(2+), and Cu(2+), but not Ni(2+), Co(2+), Mg(2+), Mn(2+), and Ca(2+), can inhibit enzyme activity. Crystal structures of these metal derivatives show that Zn(2+) and Cd(2+) bind at the alpha(1) subsite ligated by His(67), His(69), and Asp(366), while Cu(2+) at the alpha(2) subsite is chelated by His(67), His(69) and Cys(96). Unexpectedly, the crystal structure of the inactive H220A mutant displays that the endogenous Zn(2+) shifts to the alpha(3) subsite coordinated by His(67), His(69), Cys(96), and Asp(366), revealing that elimination of the beta site changes the coordination geometry of the alpha ion with an enhanced affinity. Kinetic studies of the metal ligand mutants such as C96D indicate the uniqueness of the unusual bridging cysteine and its involvement in catalysis. Therefore, the two metal-binding sites in the d aminoacylase are interactive with partially mutual exclusion, thus resulting in widely different affinities for the activation/attenuation mechanism, in which the enzyme is activated by the metal ion at the beta site, but inhibited by the subsequent binding of the second ion at the alpha site. PMID- 14736883 TI - Beta-cell-targeted overexpression of phosphodiesterase 3B in mice causes impaired insulin secretion, glucose intolerance, and deranged islet morphology. AB - The second messenger cAMP mediates potentiation of glucose-stimulated insulin release. Use of inhibitors of cAMP-hydrolyzing phosphodiesterase (PDE) 3 and overexpression of PDE3B in vitro have demonstrated a regulatory role for this enzyme in insulin secretion. In this work, the physiological significance of PDE3B-mediated degradation of cAMP for the regulation of insulin secretion in vivo and glucose homeostasis was investigated in transgenic mice overexpressing PDE3B in pancreatic beta-cells. A 2-fold overexpression of PDE3B protein and activity blunted the insulin response to intravenous glucose, resulting in reduced glucose disposal. The effects were "dose"-dependent because mice overexpressing PDE3B 7-fold failed to increase insulin in response to glucose and hence exhibited pronounced glucose intolerance. Also, the insulin secretory response to intravenous glucagon-like peptide 1 was reduced in vivo. Similarly, islets stimulated in vitro exhibited reduced insulin secretory capacity in response to glucose and glucagon-like peptide 1. Perifusion experiments revealed that the reduction specifically affected the first phase of glucose-stimulated insulin secretion. Furthermore, morphological examinations demonstrated deranged islet cytoarchitecture. In conclusion, these results are consistent with an essential role for PDE3B in cAMP-mediated regulation of insulin release and glucose homeostasis. PMID- 14736884 TI - Analysis of post-translational CCR8 modifications and their influence on receptor activity. AB - Post-translational modifications of the extracellular portions of receptors located in the cell membrane can contribute to modulating their biological activity. Using a mutagenesis approach in which single or multiple Tyr-to-Phe, Thr-to-Ala, Ser-to-Ala, and Asn-to-Gln substitutions were made at the appropriate positions, we analyzed the sulfation and glycosylation state of the murine CCR8 chemokine receptor, and the way in which these post-translational modifications affect CCR8 activity. A Y14Y15-to-F14F15 CCR8 mutant was less sulfated than the wild-type receptor. An N8-to-Q8 mutant was less glycosylated than wild-type, and a double T10T12-to-A10A12 mutant showed even less glycosylation. We established a flow cytometric analysis with an Fc-fused form of mouse CCL1 to determine precisely the ligand-binding activity of these mutants. Single mutants at amino acid positions 8, 10 or 12 bound CCL1-Fc similarly to wild-type CCR8, whereas the F14F15 double mutant was essentially inactive and the A10A12 double mutant showed about 65% of wild-type ligand-binding activity. Calcium flux activity assays were performed with these mutants, yielding results consistent with those from the ligand binding assays. These data indicate that sulfation at specific positions of the N-terminal domain of mouse CCR8 is critical for its biological activity, whereas glycosylation has a minor influence. PMID- 14736885 TI - Extracellular superoxide dismutase (EC-SOD) binds to type i collagen and protects against oxidative fragmentation. AB - The antioxidant enzyme extracellular superoxide dismutase (EC-SOD) is mainly found in the extracellular matrix of tissues. EC-SOD participates in the detoxification of reactive oxygen species by catalyzing the dismutation of superoxide radicals. The tissue distribution of the enzyme is particularly important because of the reactive nature of its substrate, and it is likely essential that EC-SOD is positioned at the site of superoxide production to prevent adventitious oxidation. EC-SOD contains a C-terminal heparin-binding region thought to be important for modulating its distribution in the extracellular matrix. This paper demonstrates that, in addition to binding heparin, EC-SOD specifically binds to type I collagen with a dissociation constant (K(d)) of 200 nm. The heparin-binding region was found to mediate the interaction with collagen. Notably, the bound EC-SOD significantly protects type I collagen from oxidative fragmentation. This expands the known repertoire of EC SOD binding partners and may play an important physiological role in preventing oxidative fragmentation of collagen during oxidative stress. PMID- 14736886 TI - Dictyostelium macroautophagy mutants vary in the severity of their developmental defects. AB - Macroautophagy is the major mechanism that eukaryotes use to recycle cellular components during stressful conditions. We have shown previously that the Atg12 Atg5 conjugation system, required for autophagosome formation in yeast, is necessary for Dictyostelium development. A second conjugation reaction, Aut7/Atg8 lipidation with phosphatidylethanolamine, as well as a protein kinase complex and a phosphatidylinositol 3-kinase complex are also required for macroautophagy in yeast. In this study, we characterize mutations in the putative Dictyostelium discoideum orthologues of budding yeast genes that are involved in one of each of these functions, ATG1, ATG6, and ATG8. All three genes are required for macroautophagy in Dictyostelium. Mutant amoebae display reduced survival during nitrogen starvation and reduced protein degradation during development. Mutations in the three genes produce aberrant development with defects of varying severity. As with other Dictyostelium macroautophagy mutants, development of atg1-1, atg6( ), and atg8(-) is more aberrant in plaques on bacterial lawns than on nitrocellulose filters. The most severe defect is observed in the atg1-1 mutant, which does not aggregate on bacterial lawns and arrests as loose mounds on nitrocellulose filters. The atg6(-) and atg8(-) mutants display almost normal development on nitrocellulose filters, producing multi-tipped aggregates that mature into small fruiting bodies. The distribution of a green fluorescent protein fusion of the autophagosome marker, Atg8, is aberrant in both atg1-1 and atg6(-) mutants. PMID- 14736887 TI - A signaling role of glutamine in insulin secretion. AB - Children with hypoglycemia due to recessive loss of function mutations of the beta-cell ATP-sensitive potassium (K(ATP)) channel can develop hypoglycemia in response to protein feeding. We hypothesized that amino acids might stimulate insulin secretion by unknown mechanisms, because the K(ATP) channel-dependent pathway of insulin secretion is defective. We therefore investigated the effects of amino acids on insulin secretion and intracellular calcium in islets from normal and sulfonylurea receptor 1 knockout (SUR1-/-) mice. Even though SUR1-/- mice are euglycemic, their islets are considered a suitable model for studies of the human genetic defect. SUR1-/- islets, but not normal islets, released insulin in response to an amino acid mixture ramp. This response to amino acids was decreased by 60% when glutamine was omitted. Insulin release by SUR1-/- islets was also stimulated by a ramp of glutamine alone. Glutamine was more potent than leucine or dimethyl glutamate. Basal intracellular calcium was elevated in SUR1-/ islets and was increased further by glutamine. In normal islets, methionine sulfoximine, a glutamine synthetase inhibitor, suppressed insulin release in response to a glucose ramp. This inhibition was reversed by glutamine or by 6 diazo-5-oxo-l-norleucine, a non-metabolizable glutamine analogue. High glucose doubled glutamine levels of islets. Methionine sulfoximine inhibition of glucose stimulated insulin secretion was associated with accumulation of glutamate and aspartate. We hypothesize that glutamine plays a critical role as a signaling molecule in amino acid- and glucose-stimulated insulin secretion, and that beta cell depolarization and subsequent intracellular calcium elevation are required for this glutamine effect to occur. PMID- 14736888 TI - Specificity of the endonuclease activity of the baculovirus alkaline nuclease for single-stranded DNA. AB - The Autographa californica multiple nucleocapsid nucleopolyhedrovirus (AcMNPV) alkaline nuclease (AN) likely participates in the maturation of virus genomes and in DNA recombination. AcMNPV AN was expressed in a recombinant baculovirus as a His -tagged fusion and obtained in pure form (*AN) or as a (6)complex with the baculoviral single-stranded DNA-binding protein LEF-3 (*AN/L3). Both AN preparations possessed potent 5' --> 3'-exonuclease and weak endonuclease activities. Mutant *AN(S146A)/L3 with a change from serine to alanine at position 146 in a conservative motif was impaired in both activities. This proved that the endonuclease is an intrinsic activity of baculovirus AN. The AN endonuclease showed specificity for single-stranded DNA and converted supercoiled plasmid DNA (replicative form I, RFI) into the open circular form (RFII) by a single strand break. Plasmid DNA relaxed with topoisomerase I was resistant to *AN/L3 indicating that the partially single-stranded regions in negatively supercoiled molecules served as targets for the endonuclease. Unwinding the supercoiled DNA with ethidium bromide also made DNA resistant to AN/L3. In reactions with nicked circular DNA (RFII), AN and AN/L3 hydrolyzed exonucleolytically the broken strand or cut endonucleolytically the intact strand at the position opposite the nick (gap). When LEF-3 was added to the assay, the balance between the exonucleolytic and endonucleolytic modes of hydrolysis shifted in favor of the exonuclease. The data suggest that the AN endonuclease may digest the intermediates in replication and recombination at positions of structural irregularities in DNA duplexes, whereas LEF-3 may further regulate processing of the intermediates by AN via the endonuclease and exonuclease pathways. PMID- 14736889 TI - Outer pore architecture of a Ca2+-selective TRP channel. AB - The TRP superfamily forms a functionally important class of cation channels related to the product of the Drosophila trp gene. TRP channels display an unusual diversity in activation mechanisms and permeation properties, but the basis of this diversity is unknown, as the structure of these channels has not been studied in detail. To obtain insight in the pore architecture of TRPV6, a Ca(2+)-selective member of the TRPV subfamily, we probed the dimensions of its pore and determined pore-lining segments using cysteine-scanning mutagenesis. Based on the permeability of the channel to organic cations, we estimated a pore diameter of 5.4 A. Mutating Asp(541), a residue involved in high affinity Ca(2+) binding, altered the apparent pore diameter, indicating that this residue lines the narrowest part of the pore. Cysteines introduced in a region preceding Asp(541) displayed a cyclic pattern of reactivity to Ag(+) and cationic methylthio-sulfanate reagents, indicative of a pore helix. The anionic methanethiosulfonate ethylsulfonate showed only limited reactivity in this region, consistent with the presence of a cation-selective filter at the outer part of the pore helix. Based on these data and on homology with the bacterial KcsA channel, we present the first structural model of a TRP channel pore. We conclude that main structural features of the outer pore, namely a selectivity filter preceded by a pore helix, are conserved between K(+) channels and TRPV6. However, the selectivity filter of TRPV6 is wider than that of K(+) channels and lined by amino acid side chains rather than main chain carbonyls. PMID- 14736890 TI - Impairment of bone healing by insulin receptor substrate-1 deficiency. AB - Insulin receptor substrate-1 (IRS-1) is an essential molecule for intracellular signaling of insulin-like growth factor (IGF)-I and insulin, both of which are potent anabolic regulators of bone and cartilage metabolism. To investigate the role of IRS-1 in bone regeneration, fracture was introduced in the tibia, and its healing was compared between wild-type (WT) mice and mice lacking the IRS-1 gene (IRS-1(-/-) mice). Among 15 IRS-1(-/-) mice, 12 remained in a non-union state even at 10 weeks after the operation, whereas all 15 WT mice showed a rigid bone union at 3 weeks. This impairment was because of the suppression of callus formation with a decrease in chondrocyte proliferation and increases in hypertrophic differentiation and apoptosis. Reintroduction of IRS-1 to the IRS-1( /-) fractured site using an adenovirus vector significantly restored the callus formation. In the culture of chondrocytes isolated from the mouse growth plate, IRS-1(-/-) chondrocytes showed less mitogenic ability and Akt phosphorylation than WT chondrocytes. An Akt inhibitor decreased the IGF-I-stimulated DNA synthesis of chondrocytes more potently in the WT culture than in the IRS-1(-/-) culture. We therefore conclude that IRS-1 deficiency impairs bone healing at least partly by inhibiting chondrocyte proliferation through the phosphatidylinositol 3-kinase/Akt pathway, and we propose that IRS-1 can be a target molecule for bone regenerative medicine. PMID- 14736891 TI - A novel NADH-linked l-xylulose reductase in the l-arabinose catabolic pathway of yeast. AB - An NADH-dependent l-xylulose reductase and the corresponding gene were identified from the yeast Ambrosiozyma monospora. The enzyme is part of the yeast pathway for l-arabinose catabolism. A fungal pathway for l-arabinose utilization has been described previously for molds. In this pathway l-arabinose is sequentially converted to l-arabinitol, l-xylulose, xylitol, and d-xylulose and enters the pentose phosphate pathway as d-xylulose 5-phosphate. In molds the reductions are NADPH-linked, and the oxidations are NAD(+)-linked. Here we show that in A. monospora the pathway is similar, i.e. it has the same two reduction and two oxidation reactions, but the reduction by l-xylulose reductase is not performed by a strictly NADPH-dependent enzyme as in molds but by a strictly NADH-dependent enzyme. The ALX1 gene encoding the NADH-dependent l-xylulose reductase is strongly expressed during growth on l-arabinose as shown by Northern analysis. The gene was functionally overexpressed in Saccharomyces cerevisiae and the purified His-tagged protein characterized. The reversible enzyme converts l xylulose to xylitol. It also converts d-ribulose to d-arabinitol but has no activity with l-arabinitol or adonitol, i.e. it is specific for sugar alcohols where, in a Fischer projection, the hydroxyl group of the C-2 is in the l configuration and the hydroxyl group of C-3 is in the d-configuration. It also has no activity with C-6 sugars or sugar alcohols. The K(m) values for l-xylulose and d-ribulose are 9.6 and 4.7 mm, respectively. To our knowledge this is the first report of an NADH-linked l-xylulose reductase. PMID- 14736892 TI - TOR complex 1 includes a novel component, Tco89p (YPL180w), and cooperates with Ssd1p to maintain cellular integrity in Saccharomyces cerevisiae. AB - The Tor1p and Tor2p kinases, targets of the therapeutically important antibiotic rapamycin, function as components of two distinct protein complexes in yeast, termed TOR complex 1 (TORC1) and TORC2. TORC1 is responsible for a wide range of rapamycin-sensitive cellular activities and contains, in addition to Tor1p or Tor2p, two highly conserved proteins, Lst8p and Kog1p. By identifying proteins that co-purify with Tor1p, Tor2p, Lst8p, and Kog1p, we have characterized a comprehensive set of protein-protein interactions that define further the composition of TORC1 as well as TORC2. In particular, we have identified Tco89p (YPL180w) and Bit61p (YJL058c) as novel components of TORC1 and TORC2, respectively. Deletion of TOR1 or TCO89 results in two specific and distinct phenotypes, (i) rapamycin-hypersensitivity and (ii) decreased cellular integrity, both of which correlate with the presence of SSD1-d, an allele of SSD1 previously associated with defects in cellular integrity. Furthermore, we link Ssd1p to Tap42p, a component of the TOR pathway that is believed to act uniquely downstream of TORC1. Together, these results define a novel connection between TORC1 and Ssd1p-mediated maintenance of cellular integrity. PMID- 14736893 TI - Stimulation of insulin fibrillation by urea-induced intermediates. AB - Fibrillar deposits of insulin cause serious problems in implantable insulin pumps, commercial production of insulin, and for some diabetics. We performed a systematic investigation of the effect of urea-induced structural perturbations on the mechanism of fibrillation of insulin. The addition of as little as 0.5 m urea to zinc-bound hexameric insulin led to dissociation into dimers. Moderate concentrations of urea led to accumulation of a partially unfolded dimer state, which dissociates into an expanded, partially folded monomeric state. Very high concentrations of urea resulted in an unfolded monomer with some residual structure. The addition of even very low concentrations of urea resulted in increased fibrillation. Accelerated fibrillation correlated with population of the partially folded intermediates, which existed at up to 8 m urea, accounting for the formation of substantial amounts of fibrils under such conditions. Under monomeric conditions the addition of low concentrations of urea slowed down the rate of fibrillation, e.g. 5-fold at 0.75 m urea. The decreased fibrillation of the monomer was due to an induced non-native conformation with significantly increased alpha-helical content compared with the native conformation. The data indicate a close-knit relationship between insulin conformation and propensity to fibrillate. The correlation between fibrillation and the partially unfolded monomer indicates that the latter is a critical amyloidogenic intermediate in insulin fibrillation. PMID- 14736896 TI - Speaking out. PMID- 14736897 TI - Speaking out. PMID- 14736899 TI - Bartonella: a new etiological agent of feline ocular disease. PMID- 14736900 TI - The use of novel stimuli as indicators of aggressive behavior in dogs. AB - To test the predictive value of a doll and an artificial hand, reactions of dogs (n=100) were compared to histories of behavior toward children. Each dog's reaction to the doll and the hand was categorized as normal, fearful, fearfully aggressive, or offensively aggressive. Sixty-five percent (n=37) of the dogs that had a normal or no reaction to the stimuli had a history of being good with children. Eighty-eight percent (n=34) of the dogs that had an aggressive reaction to the doll had a history of aggressive behavior toward a child. Dogs that were fearfully aggressive were significantly more likely to show fearful responses to the doll, and dogs that were either dominant or fearfully aggressive were more likely to exhibit aggression of the same type to the hand. The results of this study indicate that the doll and, to a lesser extent, the hand may be useful components in determining the aggressive tendencies of dogs. The results also point out the major limitations, because the false positives and false negatives are too frequent. PMID- 14736901 TI - The relationship of resting S-T segment depression to the severity of subvalvular aortic stenosis and the presence of ventricular premature complexes in the dog. AB - Electrocardiograms (ECG) from 35 dogs with subvalvular aortic stenosis (SAS) with a left ventricular outflow tract pressure gradient (PG) of > or =50 mm Hg were retrospectively evaluated for S-T segment depression (STD, > or =0.2 mV in lead II). Pressure gradient, age, heart rate (HR), and number of ventricular premature complexes (VPCs) on a 24-hour ambulatory ECG for dogs with STD were not significantly different from those for dogs without STD. The S-T segment deviation did not correlate significantly with PG, age, HR, or VPCs. The significance of STD in the dog with SAS remains uncertain. Long-term prospective studies are needed to fully understand this observation. PMID- 14736902 TI - Influence of antiarrhythmia therapy on survival times of 19 clinically healthy Doberman pinschers with dilated cardiomyopathy that experienced syncope, ventricular tachycardia, and sudden death (1985-1998). AB - Overtly healthy Doberman pinschers, having moderate to severe myocardial failure secondary to dilated cardiomyopathy, which experienced ventricular tachycardia, syncope or collapse, and sudden death were studied to determine the effect of antiarrhythmic medication on their clinical outcome. Antiarrhythmia drug therapy may have retarded sudden death in 13 treated dogs compared to the six dogs not administered antiarrhythmia drugs. PMID- 14736903 TI - Topical 0.1% tacrolimus for the treatment of discoid lupus erythematosus and pemphigus erythematosus in dogs. AB - Topical 0.1% tacrolimus was used for treatment of localized lesions associated with 10 cases of discoid lupus erythematosus (DLE) and two cases of pemphigus erythematosus (PE) either as a sole therapy (n=2) or as an adjunctive treatment (n=10). Eight of 10 dogs with DLE and both dogs with PE were improved following 8 weeks of topical application. In six of the eight dogs that improved, other medications were discontinued. No adverse effects in clinical or laboratory parameters were noted throughout the study. PMID- 14736904 TI - Comparison of body temperature in cats using a veterinary infrared thermometer and a digital rectal thermometer. AB - The purpose of this study was to determine if the temperatures obtained using a veterinary infrared (IR) thermometer agreed with a digital rectal thermometer in a group of research cats, half of which had transient fevers. The thermometers were weakly correlated (r=0.62). The mean difference was 0.13 degrees F (0.07 degrees C), and the limits of agreement were 2.6 degrees F (1.43 degrees C) and 2.5 degrees F (-1.36 degrees C), which were unacceptable for clinical purposes. The results of this study indicate that, while the IR thermometer was easy to use, it cannot be used interchangeably with the rectal thermometer. PMID- 14736905 TI - Vaccine site-associated sarcoma and malignant lymphoma in cats: a report of six cases (1997-2002). AB - Six cats developed malignant lymphoma 3 to 45 months after treatment for vaccine site-associated sarcoma. During the same time period, 184 cats were evaluated in the teaching hospital for vaccine site-associated sarcomas. Feline vaccine site associated sarcoma is not believed to be associated with feline leukemia virus (FeLV) infection. Five of six cats were negative by enzyme-linked immunosorbent assay for FeLV antigens at the times of diagnosis of both sarcoma and lymphoma, and no cats were infected with feline immunodeficiency virus. PMID- 14736906 TI - Chronic diaphragmatic hernia in 34 dogs and 16 cats. AB - Medical records of 34 dogs and 16 cats undergoing surgical repair of diaphragmatic hernia of >2 weeks' duration were reviewed, and long-term follow-up information was obtained. The most common clinical signs were dyspnea and vomiting; however, many of the animals were presented for nonspecific signs such as anorexia, lethargy, and weight loss. Thoracic radiographs revealed evidence of diaphragmatic hernia in only 66% of the animals, and additional imaging tests were often needed to confirm the diagnosis. Thirty-six hernias were repaired through a midline laparotomy; 14 required a median sternotomy combined with a laparotomy. In 14 animals, division of mature adhesions of the lungs or diaphragm to the herniated organs was necessary to permit reduction of the hernia. Fourteen animals required resection of portions of the lungs, liver, or intestine. All hernias were sutured primarily without the use of tissue flaps or mesh implants. Twenty-one of the animals developed transient complications in the postoperative period; the most common of these was pneumothorax. The mortality rate was 14%. Thirty-four (79%) of the animals that were discharged from the hospital had complete resolution of clinical signs, and none developed evidence of recurrent diaphragmatic hernia during the follow-up period. Nine were lost to follow-up. PMID- 14736907 TI - Intravesical instillation of dilute formalin for the treatment of severe hemorrhagic emphysematous cystitis in a diabetic dog. AB - Intravesical formalin is a known treatment for control of hemorrhagic cystitis caused by multiple etiologies in humans and dogs. This case report documents the successful use of intravesical formalin for the treatment of severe hemorrhagic cystitis that occurred secondary to emphysematous cystitis in a diabetic dog. In addition, a review of emphysematous cystitis and the use of intravesical formalin in human and veterinary medicine is discussed. Formalin instillation into the urinary bladder is an option for life-threatening, refractory cases of hemorrhagic cystitis; but clinicians must be familiar with the proper technique and be aware of potential complications prior to its use. PMID- 14736908 TI - Pancreatolithiasis and pancreatic pseudobladder associated with pancreatitis in a cat. AB - Pancreatolithiasis has been documented to occur naturally in humans and cattle. It has been associated with chronic pancreatitis in humans, and, when found, it may signify the presence of chronic pancreatic disease. This is the first report of a case involving a cat that had both an apparent obstruction with pancreatolithiasis as well as concurrent evidence of chronic pancreatic changes on histopathological evaluation. Additionally, this case documents the presence of a suspected congenital abnormality of a feline exocrine pancreas. PMID- 14736909 TI - Electrolyte disturbances and cardiac arrhythmias in a dog following pamidronate, calcitonin, and furosemide administration for hypercalcemia of malignancy. AB - A 13-year-old dog was diagnosed with hypercalcemia of malignancy associated with adenocarcinoma of the anal sacs. Hypercalcemia was treated with intravenous (IV) 0.9% sodium chloride (NaCl), furosemide, calcitonin, and pamidronate. Hypomagnesemia was documented by 72 hours following a single, IV dose of pamidronate. The dog subsequently underwent surgery to remove the primary tumors, and multiple cardiac arrhythmias occurred during anesthesia. This case documents electrolyte abnormalities in a dog following treatment with pamidronate in conjunction with other therapies used to manage hypercalcemia. The authors postulate that hypomagnesemia may have contributed to the arrhythmias that occurred during anesthesia. Electrolyte abnormalities should be anticipated and corrected following pamidronate therapy in canine patients. PMID- 14736910 TI - Ureteral mast cell tumor in a dog. AB - A 6-year-old, castrated male, mixed-breed dog was diagnosed with partial unilateral ureteral obstruction secondary to a ureteral mass. The ureteral mass was surgically resected, and an ureteroneocystostomy was performed. Histopathology of the ureteral mass was consistent with a poorly differentiated mast cell tumor (MCT). The patient recovered well but was euthanized 5 months postoperatively for central nervous system signs. A choroid plexus tumor was diagnosed during necropsy examination. There was no evidence of recurrence or dissemination of the ureteral MCT. Extracutaneous MCTs are rare in dogs, and primary MCT associated with the urinary tract has not previously been reported in the veterinary literature. PMID- 14736911 TI - Reconstruction of a bilateral hypoplastic soft palate in a cat. AB - A 2-year-old, female spayed, domestic shorthair cat was presented with a history of dyspnea, upper respiratory congestion, and aspiration. Physical examination revealed a bilaterally hypoplastic soft palate resembling a uvula. A novel surgical technique was performed to reconstruct the soft palate, utilizing both a hard palate-derived mucoperiosteal flap combined with two pharyngeal wall derived, random-pattern mucosal flaps. This is the first report of a bilaterally hypoplastic soft palate in the cat, as well as a description of a new surgical method for soft palate reconstruction. PMID- 14736912 TI - Lipid-control allies. PMID- 14736913 TI - Getting connected. PMID- 14736914 TI - Dear old dad. AB - The origin and frequency of spontaneous mutations that occur with age in humans have been a topic of intense discussion. The mechanisms by which spontaneous mutations arise depend on the parental germ line in which a mutation occurs. In general, paternal mutations are more likely than maternal mutations to be base substitutions. This is likely due to the larger number of germ cell divisions in spermatogenesis than in oogenesis. Maternal mutations are more often chromosomal abnormalities. Advanced parental age seems to influence some mutations, although it is not a factor in the creation of others. In this review, we focus on patterns of paternal bias and age dependence of mutations in different genetic disorders, and the various mechanisms by which these mutations arise. We also discuss recent data on age and the frequency of these mutations in the human male germ line and the impact of these data on this field of research. PMID- 14736915 TI - Giant plasmid-encoded polyketide synthases produce the macrolide toxin of Mycobacterium ulcerans. AB - Mycobacterium ulcerans (MU), an emerging human pathogen harbored by aquatic insects, is the causative agent of Buruli ulcer, a devastating skin disease rife throughout Central and West Africa. Mycolactone, an unusual macrolide with cytotoxic and immunosuppressive properties, is responsible for the massive s.c. tissue destruction seen in Buruli ulcer. Here, we show that MU contains a 174-kb plasmid, pMUM001, bearing a cluster of genes encoding giant polyketide synthases (PKSs), and polyketide-modifying enzymes, and demonstrate that these are necessary and sufficient for mycolactone synthesis. This is a previously uncharacterized example of plasmid-mediated virulence in a Mycobacterium, and the emergence of MU as a pathogen most likely reflects the acquisition of pMUM001 by horizontal transfer. The 12-membered core of mycolactone is produced by two giant, modular PKSs, MLSA1 (1.8 MDa) and MLSA2 (0.26 MDa), whereas its side chain is synthesized by MLSB (1.2 MDa), a third modular PKS highly related to MLSA1. There is an extreme level of sequence identity within the different domains of the MLS cluster (>97% amino acid identity), so much so that the 16 ketosynthase domains seem functionally identical. This is a finding of significant consequence for our understanding of polyketide biochemistry. Such detailed knowledge of mycolactone will further the investigation of its mode of action and the development of urgently needed therapeutic strategies to combat Buruli ulcer. PMID- 14736916 TI - Gene replacement reveals that p115/SNARE interactions are essential for Golgi biogenesis. AB - Functional characterization of protein interactions in mammalian systems has been hindered by the inability to perform complementation analyses in vivo. Here, we use functional replacement of the vesicle docking protein p115 to separate its essential from its nonessential interactions. p115 is required for biogenesis of the Golgi apparatus, but it is unclear whether its mechanism of action requires its golgin and/or SNARE interactions. Short interfering RNA-mediated knockdown of p115 induced extensive Golgi fragmentation and impaired secretory traffic. Reassembly of a structurally and functionally normal Golgi occurred on expression of a p115 homologue not recognized by the short interfering RNA. Strikingly, versions of p115 lacking its phosphorylation site and the golgin-binding domains also restored the Golgi apparatus in cells lacking endogenous p115. In contrast, the p115 SNARE-interacting domain was required for Golgi biogenesis. This suggests that p115 acts directly, rather than via a tether, to catalyze trans SNARE complex formation preceding membrane fusion. PMID- 14736917 TI - Calmodulin phosphorylation and modulation of endothelial nitric oxide synthase catalysis. AB - The endothelial NO synthase (eNOS) is regulated by diverse protein kinase pathways, yet eNOS activity ultimately depends on the ubiquitous calcium regulatory protein calmodulin (CaM). In these studies, we establish that CaM itself undergoes phosphorylation in endothelial cells and that CaM phosphorylation attenuates eNOS activation. Using [(32)P]orthophosphoric acid biosynthetic labeling, we found that CaM is a phosphoprotein in bovine aortic endothelial cells (BAEC) and that the kinase CK2 promotes CaM phosphorylation in BAEC. Phosphorylation of CaM by purified CK2 in vitro reduces the V(max) of immunopurified eNOS by a factor of 2 but has no effect on the K(A) for CaM or calcium. Additionally, [(32)P]orthophosphoric acid biosynthetic labeling of mutant CaM-transfected BAEC revealed that phosphorylation of Ser-81 to alanine mutant CaM ("phosphonull" S81A mutant) is dramatically reduced relative to WT, whereas phosphorylation of the "phosphomimetic" Ser-81 to aspartate (S81D) mutant is unchanged. Further studies using Escherichia coli-expressed and phenyl Sepharose-purified CaM mutants revealed that the S81A mutation abrogates in vitro CK2-mediated phosphorylation of CaM, whereas phosphorylation of the S81D CaM mutant by CK2 is preserved. Additionally, we found that the phosphomimetic S101D CaM mutant is impaired in its ability to activate eNOS. Taken together, these results suggest that phosphorylation of CaM inhibits eNOS catalysis and proceeds in a hierarchical manner, initially requiring phosphorylation of the CaM Ser-81 residue. We conclude that CaM phosphorylation may represent a unique pathway in the regulation of eNOS signaling and thereby may play a role in modulating NO dependent vascular responses. PMID- 14736918 TI - Genomic identification of direct target genes of LEAFY. AB - The switch from vegetative to reproductive development in plants necessitates a switch in the developmental program of the descendents of the stem cells in the shoot apical meristem. Genetic and molecular investigations have demonstrated that the plant-specific transcription factor and meristem identity regulator LEAFY (LFY) controls this developmental transition by inducing expression of a second transcription factor, APETALA1, and by regulating the expression of additional, as yet unknown, genes. Here we show that the additional LFY targets include the APETALA1-related factor, CAULIFLOWER, as well as three transcription factors and two putative signal transduction pathway components. These genes are up-regulated by LFY even when protein synthesis is inhibited and, hence, appear to be direct targets of LFY. Supporting this conclusion, cis-regulatory regions upstream of these genes are bound by LFY in vivo. The newly identified LFY targets likely initiate the transcriptional changes that are required for the switch from vegetative to reproductive development in Arabidopsis. PMID- 14736919 TI - Duplication, coclustering, and selection of human Alu retrotransposons. AB - Alu and L1 are families of non-LTR retrotransposons representing approximately equal 30% of the human genome. Genomic distributions of young Alu and L1 elements are quite similar, but over time, Alu densities in GC-rich DNA increase in comparison with L1 densities. Here we analyze two processes that may contribute to this phenomenon. First, DNA duplications in the human genome occur more frequently in Alu- and GC-rich than in AT-rich chromosomal regions. Second, most Alu elements tend to be coclustered with each other, but recently retroposed elements are likely to be inserted outside the existing clusters. These "stand alone" elements appear to be rapidly eliminated from the genome. We also report that over time, the densities of recently retroposed Alu families on chromosome Y decline rapidly, whereas Alu densities on chromosome X increase relative to autosomal densities. We propose that these changes in the chromosomal proportions of Alu densities and the elimination of stand-alone Alus represent the same process of paternal Alu selection. We also propose that long-term Alu accumulation in GC-rich DNA is associated with DNA duplication initiated by elevated recombinogenic activities in Alu clusters. PMID- 14736920 TI - Multiple evidence for nucleotide metabolism in the chloroplast thylakoid lumen. AB - The apparatus of photosynthetic energy conversion in chloroplasts is quite well characterized with respect to structure and function. Light-driven electron transport in the thylakoid membrane is coupled to synthesis of ATP, used to drive energy-dependent metabolic processes in the stroma and the outer surface of the thylakoid membrane. The role of the inner (luminal) compartment of the thylakoids has, however, remained largely unknown although recent proteomic analyses have revealed the presence of up to 80 different proteins. Further, there are no reports concerning the presence of nucleotides in the thylakoid lumen. Here, we bring three lines of experimental evidence for nucleotide-dependent processes in this chloroplast compartment. (i) The thylakoid lumen contains a protein of 17.2 kDa, catalyzing the transfer of the gamma-phosphate group from ATP to GDP, proposed to correspond to the nucleoside diphosphate kinase III. (ii) The 33-kDa subunit of photosystem II, bound to the luminal side of the thylakoid membrane and associated with the water-splitting process, can bind GTP. (iii) The thylakoid membrane contains a nucleotide transport system that is suggested to be associated with a 36.5-kDa nucleotide-binding protein. Our results imply, against current dogmas, that the thylakoid lumen contains nucleotides, thereby providing unexpected aspects on this chloroplast compartment from a metabolic and regulatory perspective and expanding its functional significance beyond a pure bioenergetic function. PMID- 14736921 TI - Three facets of sexual differentiation. PMID- 14736922 TI - Poxvirus zoonoses--putting pocks into context. PMID- 14736923 TI - Financial conflicts of interest and the NIH. PMID- 14736924 TI - Health coverage in the states--Maine's plan for universal access. PMID- 14736925 TI - Discordant sexual identity in some genetic males with cloacal exstrophy assigned to female sex at birth. AB - BACKGROUND: Cloacal exstrophy is a rare, complex defect of the entire pelvis and its contents that occurs during embryogenesis and is associated with severe phallic inadequacy or phallic absence in genetic males. For about 25 years, neonatal assignment to female sex has been advocated for affected males to overcome the issue of phallic inadequacy, but data on outcome remain sparse. METHODS: We assessed all 16 genetic males in our cloacal-exstrophy clinic at the ages of 5 to 16 years. Fourteen underwent neonatal assignment to female sex socially, legally, and surgically; the parents of the remaining two refused to do so. Detailed questionnaires extensively evaluated the development of sexual role and identity, as defined by the subjects' persistent declarations of their sex. RESULTS: Eight of the 14 subjects assigned to female sex declared themselves male during the course of this study, whereas the 2 raised as males remained male. Subjects could be grouped according to their stated sexual identity. Five subjects were living as females; three were living with unclear sexual identity, although two of the three had declared themselves male; and eight were living as males, six of whom had reassigned themselves to male sex. All 16 subjects had moderate-to-marked interests and attitudes that were considered typical of males. Follow-up ranged from 34 to 98 months. CONCLUSIONS: Routine neonatal assignment of genetic males to female sex because of severe phallic inadequacy can result in unpredictable sexual identification. Clinical interventions in such children should be reexamined in the light of these findings. PMID- 14736926 TI - The detection of monkeypox in humans in the Western Hemisphere. AB - BACKGROUND: During May and June 2003, an outbreak of febrile illness with vesiculopustular eruptions occurred among persons in the midwestern United States who had had contact with ill pet prairie dogs obtained through a common distributor. Zoonotic transmission of a bacterial or viral pathogen was suspected. METHODS: We reviewed medical records, conducted interviews and examinations, and collected blood and tissue samples for analysis from 11 patients and one prairie dog. Histopathological and electron-microscopical examinations, microbiologic cultures, and molecular assays were performed to identify the etiologic agent. RESULTS: The initial Wisconsin cases evaluated in this outbreak occurred in five males and six females ranging in age from 3 to 43 years. All patients reported having direct contact with ill prairie dogs before experiencing a febrile illness with skin eruptions. We found immunohistochemical or ultrastructural evidence of poxvirus infection in skin-lesion tissue from four patients. Monkeypox virus was recovered in cell cultures of seven samples from patients and from the prairie dog. The virus was identified by detection of monkeypox-specific DNA sequences in tissues or isolates from six patients and the prairie dog. Epidemiologic investigation suggested that the prairie dogs had been exposed to at least one species of rodent recently imported into the United States from West Africa. CONCLUSIONS: Our investigation documents the isolation and identification of monkeypox virus from humans in the Western Hemisphere. Infection of humans was associated with direct contact with ill prairie dogs that were being kept or sold as pets. PMID- 14736927 TI - Cisplatin-based adjuvant chemotherapy in patients with completely resected non small-cell lung cancer. AB - BACKGROUND: On the basis of a previous meta-analysis, the International Adjuvant Lung Cancer Trial was designed to evaluate the effect of cisplatin-based adjuvant chemotherapy on survival after complete resection of non-small-cell lung cancer. METHODS: We randomly assigned patients either to three or four cycles of cisplatin-based chemotherapy or to observation. Before randomization, each center determined the pathological stages to include, its policy for chemotherapy (the dose of cisplatin and the drug to be combined with cisplatin), and its postoperative radiotherapy policy. The main end point was overall survival. RESULTS: A total of 1867 patients underwent randomization; 36.5 percent had pathological stage I disease, 24.2 percent stage II, and 39.3 percent stage III. The drug allocated with cisplatin was etoposide in 56.5 percent of patients, vinorelbine in 26.8 percent, vinblastine in 11.0 percent, and vindesine in 5.8 percent. Of the 932 patients assigned to chemotherapy, 73.8 percent received at least 240 mg of cisplatin per square meter of body-surface area. The median duration of follow-up was 56 months. Patients assigned to chemotherapy had a significantly higher survival rate than those assigned to observation (44.5 percent vs. 40.4 percent at five years [469 deaths vs. 504]; hazard ratio for death, 0.86; 95 percent confidence interval, 0.76 to 0.98; P<0.03). Patients assigned to chemotherapy also had a significantly higher disease-free survival rate than those assigned to observation (39.4 percent vs. 34.3 percent at five years [518 events vs. 577]; hazard ratio, 0.83; 95 percent confidence interval, 0.74 to 0.94; P<0.003). There were no significant interactions with prespecified factors. Seven patients (0.8 percent) died of chemotherapy-induced toxic effects. CONCLUSIONS: Cisplatin-based adjuvant chemotherapy improves survival among patients with completely resected non-small-cell lung cancer. PMID- 14736928 TI - Tanapox infection in a college student. PMID- 14736929 TI - Sex determination and differentiation. PMID- 14736931 TI - Images in clinical medicine. True hermaphroditism. PMID- 14736930 TI - Multidisciplinary management of lung cancer. PMID- 14736932 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 3-2004. A 57-year-old man with invasive transitional-cell carcinoma of the bladder. PMID- 14736933 TI - Adjuvant chemotherapy for lung cancer--a new standard of care. PMID- 14736934 TI - Paying physicians for high-quality care. PMID- 14736935 TI - The danger within. PMID- 14736936 TI - Sirolimus-eluting coronary stents. PMID- 14736937 TI - Extending the interval between cervical-cancer screenings. PMID- 14736938 TI - Schizophrenia, drug therapy, and monitoring. PMID- 14736939 TI - Case 20-2003: Gaucher's disease. PMID- 14736940 TI - Intraabdominal pregnancy after hysterectomy. PMID- 14736941 TI - A 38-year history of natural-killer-cell lymphoma. PMID- 14736942 TI - What made Hanno Buddenbrook sick? PMID- 14736943 TI - Images in clinical medicine. Pneumatosis intestinalis and portal venous gas. PMID- 14736944 TI - Proteinuria and interstitial injury. PMID- 14736945 TI - Deranged removal of apoptotic cells: its role in the genesis of lupus. PMID- 14736946 TI - Cyclosporin A toxicity, and more: vascular endothelial growth factor (VEGF) steps forward. PMID- 14736947 TI - Erythropoiesis-stimulating agents and antibody-mediated pure red-cell aplasia: here are we now and where do we go from here? PMID- 14736948 TI - Which dialyser membrane to choose? PMID- 14736949 TI - Monoclonal antibodies in renal transplantation: old and new. PMID- 14736950 TI - Proteinuria after renal transplantation: pathogenesis and management. PMID- 14736951 TI - Stenosis and thrombosis in haemodialysis fistulae and grafts: the radiologist's point of view. PMID- 14736952 TI - Stenosis and thrombosis in haemodialysis fistulae and grafts: the surgeon's point of view. PMID- 14736953 TI - Dexamethasone regulates AP-1 to repress TNF-alpha induced MCP-1 production in human glomerular endothelial cells. AB - BACKGROUND: Glomerular endothelial cells play a role in the pathogenesis of glomerulonephritis by producing chemotactic factors. We investigated the role of NF-kappa B and AP-1 in tumour necrosis factor alpha (TNF-alpha) induced monocyte chemoattractant protein 1 (MCP-1) production in cultured human glomerular endothelial cells (HGEC). We also examined whether or not these processes could be modified by glucocorticoid. METHODS: MCP-1 protein and mRNA levels were measured by ELISA and northern blot. NF-kappa B and AP-1 binding activity were assessed by electrophoretic mobility shift assay. Cytosolic I kappa B alpha and nuclear p65 protein were evaluated by western blot. For specific inhibition of NF kappa B or AP-1, we used a decoy oligodeoxynucleotide. RESULTS: TNF-alpha (10 ng/ml) increased MCP-1 mRNA expression in HGEC and also the release of MCP-1 protein into culture media. These effects could be partially inhibited by dexamethasone (10 nM). TNF-alpha induced MCP-1 production appeared to be NF-kappa B and AP-1 interdependent, based on the following results. (i) TNF-alpha increased NF-kappa B and AP-1 binding activity. (ii) Both NF-kappa B decoy oligodeoxynucleotide and AP-1 decoy oligodeoxynucleotide partially suppressed TNF alpha induced MCP-1 mRNA expression. On the other hand, dexamethasone decreased TNF-alpha induced DNA-binding activity of AP-1 without an effect on the DNA binding activity of NF-kappa B, cytosolic I kappa B alpha degradation or p65 nuclear translocation. CONCLUSIONS: These data demonstrate that while TNF-alpha induced MCP-1 production is mediated by the cooperative action of NF-kappa B and AP-1 in HGEC, dexamethasone represses TNF-alpha induced MCP-1 production via suppression of AP-1 binding activity. PMID- 14736954 TI - Renal processing of serum proteins in an albumin-deficient environment: an in vivo study of glomerulonephritis in the Nagase analbuminaemic rat. AB - BACKGROUND: Plasma albumin has been considered as important for governing glomerular permselectivity as well as being tubulotoxic in proteinuric states. The purpose of this study was to examine glomerular permselectivity and protein clearance of plasma albumin-deficient Nagase analbuminaemic rats (NAR) in normal and proteinuric states associated with puromycin aminonucleoside nephrosis (PAN) and anti-glomerular basement membrane glomerulonephritis (anti-GBM GN) and to compare the results with those of previous studies using Sprague-Dawley rats. METHODS: Glomerular permselectivity was measured using tritium-labelled polydisperse Ficoll. In vivo fractional clearance (FC) of albumin, transferrin and immunoglobulin G was measured to include both intact and degraded forms of filtered material. Endogenous protein clearance was analysed using two dimensional electrophoresis in combination with matrix-assisted laser desorption ionization (MALDI) mass spectrometry. RESULTS: FCs of proteins and Ficoll in control NAR were similar to those found in Sprague-Dawley rats. Despite the lack of serum albumin in NAR, proteinuria and morphological changes observed were also similar to those found in Sprague-Dawley rats, with total protein excretion increasing 6-fold in PAN rats and 4-fold in anti-GBM GN rats with respect to controls. Two-dimensional electrophoresis in combination with MALDI mass spectrometry identified the major proteins being excreted as transferrin and a group of mildly acidic proteins in the MW range 40-50 kDa, namely antithrombin III, kininogen, alpha-1-antiproteinase, haemopexin and vitamin D-binding protein. CONCLUSIONS: Both diseases exhibited similar effects to those observed in Sprague Dawley rats despite the lack of serum albumin, including inhibition of renal protein degradation. The net changes in protein FC, particularly in the range of radii of 36-55 A, could not be accounted for by changes in size selectivity as Ficoll FC was little affected by the disease states. This emphasizes the need to reassess the relative importance of changes in renal tubular handling vs changes in glomerular capillary barrier in proteinuric states. These studies also demonstrate that albumin is not a critical factor in governing glomerular permselectivity or proteinuria. PMID- 14736955 TI - Mitochondrial DNA and its respiratory chain products are defective in doxorubicin nephrosis. AB - BACKGROUND: Doxorubicin induces a self-perpetuating nephropathy characterized by early glomerular and late-onset tubular lesions in rats. We investigated the potential role of mitochondrial injury in the onset of these lesions. METHODS: Rats were treated with intravenous doxorubicin (1 mg kg(-1) week(-1)) for 7 weeks and were sacrificed either 1 week ('short-term') or 30 weeks ('long-term') following the last dose. Additional rats received a single dose either 6 days or 2 h prior to euthanasia. All rats were killed at 48 weeks of age. Glomerular and tubular injury was monitored and correlated to the activity or expression of respiratory chain components. Finally, we quantified both nuclear and mitochondrial DNA (mtDNA) as well as superoxide production and the 4834 base pair 'common' mtDNA deletion. RESULTS: The 'long-term' group had significant glomerular and tubular lesions, depressed activities of mtDNA-encoded NADH dehydrogenase and cytochrome-c oxidase (COX) and increased citrate synthase activity. In addition, expression of the mtDNA-encoded COX subunit I was reduced and mtDNA levels were decreased. In 'short-term' rats, there were fewer tubular lesions, but similar numbers of glomerular lesions activity. Among all animals, glomerular and tubular injury were inversely correlated with mtDNA levels, mtDNA encoded respiratory chain activities and with the expression of the mtDNA-encoded respiratory chain subunit COX-I. Injury was positively correlated with superoxide production and the activities of nucleus-encoded mitochondrial or cytoplasmic enzymes. Kidneys from the 'long-term' group showed more mtDNA deletions than in 'short-term' animals and these were not observed in the other groups. CONCLUSIONS: These results suggest an important role for quantitative and qualitative mtDNA alterations through the reduction of mtDNA-encoded respiratory chain function and induction of superoxide in doxorubicin-induced renal lesions. PMID- 14736956 TI - Impact of the supplementation of kidney mass on blood pressure and progression of kidney disease. AB - BACKGROUND: To test the hypothesis that nephron mass is an independent determinant of arterial pressure, the effects of augmenting renal mass by isograft transplantation were studied in the model of secondary hypertension. METHODS: The effects of isograft transplantation or sham operation on blood pressure, proteinuria, remnant kidney mass, glomerular filtration rate and glomerulosclerosis were assessed in 5/6 nephrectomized (5/6 NPX) rats. RESULTS: Systolic blood pressure was lowered on average by approximately 35 mmHg and glomerular hyperfiltration was attenuated in the remnant kidneys of transplant recipients. Markedly lower urinary protein excretion rates and glomerulosclerosis scores in the remnant kidney accompanied these supplemental transplants to values roughly one-third of those from sham-operated rats. CONCLUSIONS: The data show that reduced renal mass per se is the major factor in the development and maintenance of arterial hypertension and glomerular injury in 5/6 NPX rats and these changes can be reversed by supplementing renal mass. The data provide strong support for the notion that renal mass is a significant, independent determinant of arterial pressure. PMID- 14736957 TI - Chronic renal denervation prevents glomerular hyperfiltration in diabetic rats. AB - BACKGROUND: The increase in glomerular filtration rate (GFR) induced by amino acid infusion is attenuated in rats with chronic renal denervation. The aim of the present study was to investigate whether renal denervation abrogates glomerular hyperfiltration occurring in the early state of diabetes mellitus. METHODS: Sprague-Dawley rats were subjected to bilateral renal denervation before induction of diabetes mellitus (DM) by streptozotocin. Clearance experiments were performed 2 weeks after onset of moderate DM. Glomerular volume was estimated following paraformaldehyde fixation in rat kidney slices from measurement of cross-sectional area of Bowman's capsule. RESULTS: GFR in non-diabetic rats with intact nerves (CON-INN) was 0.82+/-0.03 ml.min(-1).100 g(-1) body weight. Diabetic animals with innervated kidneys presented a significant glomerular hyperfiltration (1.13+/-0.03 ml.min(-1).100 g(-1) body weight), while renal denervation in diabetic rats lowered GFR towards levels of CON-INN (0.88+/-0.03 ml.min(-1).100 g(-1) body weight). Estimated glomerular volume amounted to 0.69+/ 0.03.10(6) micro m(3) in the CON-INN group and was significantly higher in diabetic animals with intact renal nerves (0.86+/-0.04.10(6) microm(3)). Interestingly, renal denervation prevented the glomerular enlargement due to DM. CONCLUSIONS: Renal nerves appear to be significantly involved in the mediation of glomerular hyperfiltration in experimental DM. If the kidney is prevented from sympathetic nerve stimulation, structural changes due to early diabetic nephropathy, i.e. glomerular enlargement, are abolished. PMID- 14736958 TI - Erythropoietin protects against ischaemic acute renal injury. AB - BACKGROUND: Erythropoietin (EPO) has recently been shown to exert important cytoprotective and anti-apoptotic effects in experimental brain injury and cisplatin-induced nephrotoxicity. The aim of the present study was to determine whether EPO administration is also renoprotective in both in vitro and in vivo models of ischaemic acute renal failure. METHODS: Primary cultures of human proximal tubule cells (PTCs) were exposed to either vehicle or EPO (6.25-400 IU/ml) in the presence of hypoxia (1% O(2)), normoxia (21% O(2)) or hypoxia followed by normoxia for up to 24 h. The end-points evaluated included cell apoptosis (morphology and in situ end labelling [ISEL], viability [lactate dehydrogenase (LDH release)], cell proliferation [proliferating cell nuclear antigen (PCNA)] and DNA synthesis (thymidine incorporation). The effects of EPO pre-treatment (5000 U/kg) on renal morphology and function were also studied in rat models of unilateral and bilateral ischaemia-reperfusion (IR) injury. RESULTS: In the in vitro model, hypoxia (1% O(2)) induced a significant degree of PTC apoptosis, which was substantially reduced by co-incubation with EPO at 24 h (vehicle 2.5+/-0.5% vs 25 IU/ml EPO 1.8+/-0.4% vs 200 IU/ml EPO 0.9+/-0.2%, n = 9, P<0.05). At high concentrations (400 IU/ml), EPO also stimulated thymidine incorporation in cells exposed to hypoxia with or without subsequent normoxia. LDH release was not significantly affected. In the unilateral IR model, EPO pre treatment significantly attenuated outer medullary thick ascending limb (TAL) apoptosis (EPO 2.2+/-1.0% of cells vs vehicle 6.5+/-2.2%, P<0.05, n = 5) and potentiated mitosis (EPO 1.1+/-0.3% vs vehicle 0.5+/-0.3%, respectively, P<0.05) within 24 h. EPO-treated rats exhibited enhanced PCNA staining within the proximal straight tubule (6.9+/-0.7% vs vehicle 2.4+/-0.5% vs sham 0.3+/-0.2%, P<0.05), proximal convoluted tubule (2.3+/-0.6% vs vehicle 1.1+/-0.3% vs sham 1.2+/-0.3%, P<0.05) and TAL (4.7+/-0.9% vs vehicle 0.6+/-0.3% vs sham 0.3+/-0.2%, P<0.05). The frequency of tubular profiles with luminal cast material was also reduced (32.0+/-1.6 vs vehicle 37.0+/-1.3%, P = 0.05). EPO-treated rats subjected to bilateral IR injury exhibited similar histological improvements to the unilateral IR injury model, as well as significantly lower peak plasma creatinine concentrations than their vehicle-treated controls (0.04+/-0.01 vs 0.21+/-0.08 mmol/l, respectively, P<0.05). EPO had no effect on renal function in sham operated controls. CONCLUSIONS: The results suggest that, in addition to its well known erythropoietic effects, EPO inhibits apoptotic cell death, enhances tubular epithelial regeneration and promotes renal functional recovery in hypoxic or ischaemic acute renal injury. PMID- 14736959 TI - Outcome in glomerulonephritis due to systemic small vessel vasculitis: effect of functional status and non-vasculitic co-morbidity. AB - BACKGROUND: Previous studies in systemic vasculitis have defined a number of disease-specific factors including histological parameters and clinical vasculitis activity scores as outcome predictors. This study evaluates the previously neglected role of non-vasculitic factors in determining prognosis in this condition. METHODS: We performed a retrospective study of all patients with a diagnosis of small vessel vasculitis (SVV) presenting with renal impairment to our service over a 12-year period. RESULTS: Eighty-six patients were studied (median age 63.5 years, 64% male). Mean plasma creatinine at presentation was 533 micro M (6.3 mg/dl). Forty-seven patients (55%) required immediate dialysis, 21 (45%) recovered renal function on treatment. The presence of crescent fibrosis (P < 0.05) and interstitial fibrosis (P < 0.01) were significantly associated with a failure to recover renal function. There was a trend towards an increased relapse rate in those with a persistently positive ANCA result or a rising titre. Twelve month patient survival was 85.5% and 5-year survival was 63%. Factors independently associated with mortality were Karnofsky performance score at diagnosis (P < 0.00001), intensity of immunosuppressive treatment (P = 0.0007) and vasculitis classification (P = 0.009). Non-vasculitic co-morbidity was not independently associated with mortality. Patients who were, or became after treatment, dialysis-independent had a significant survival advantage (5-year survival 83 vs 42%, P = 0.001). CONCLUSIONS: Non-vasculitic factors, particularly functional status as indicated by the Karnofsky performance score, play a major role in determining prognosis in SVV with renal involvement and should be an integral component of the decision making process when planning therapy, and in comparing outcomes between centres. PMID- 14736960 TI - The role of flow cytometric ANCA detection in screening for acute pauci-immune crescentic glomerulonephritis. AB - BACKGROUND: Most cases of pauci-immune crescentic glomerulonephritis (PICGN) are associated with serum anti-neutrophil cytoplasmic antibodies (ANCA). This article studied the sensitivity and specificity of serum ANCA, determined by flow cytometry and indirect immunofluorescence (IIF), to identify patients with acute PICGN. METHODS: 577 adults presenting for first biopsy of their native kidneys with serum taken for ANCA (flow cytometry and IIF) determination were studied. A positive ANCA was defined using a flow cytometric ANCA assay as a screening test, followed by a slide-based indirect IIF technique. Pathological confirmation of acute PICGN was used to assess the sensitivity and specificity of this combined approach and its positive predictive value (PPV) and negative predictive value (NPV) in patients presenting for renal biopsy due to abnormal urinary sediment. RESULTS: Forty-nine patients were found to have acute PICGN on renal biopsy. Of these 47 were ANCA positive (sensitivity 95.9%). Overall 93 of the renal biopsy patients were ANCA positive, (specificity 91.3%). A further seven patients (two ANCA positive) had advanced sclerosing disease consistent with PICGN but without evidence of current disease activity. The PPV and NPV of ANCA, assessed by flow cytometry and slide IIF, in predicting that patients presenting with undifferentiated renal disease would have acute PICGN was 50.5 and 99.8%, respectively. CONCLUSIONS: Flow cytometric screening of serum for ANCA in patients undergoing renal biopsy has a high NPV for determining those with acute PICGN. It may provide a rapid, simple screening test for this lesion in laboratories using diagnostic flow cytometry and may complement IIF/ELISA in evaluating ANCA positive patients. PMID- 14736961 TI - Characterization of a highly polymorphic marker adjacent to the SLC4A1 gene and of kidney immunostaining in a family with distal renal tubular acidosis. AB - BACKGROUND: Mutations in the human SLC4A1 (AE1/band 3) gene are associated with hereditary spherocytic anaemia and with distal renal tubular acidosis (dRTA). The molecular diagnosis of AE1 mutations has been complicated by the absence of highly polymorphic genetic markers, and the pathogenic mechanisms of some dRTA associated AE1 mutations remain unclear. Here, we characterized a polymorphic dinucleotide repeat close to the human AE1 gene and performed an immunocytochemical study of kidney tissue from a patient with inherited dRTA with a defined AE1 mutation. METHODS: One CA repeat region was identified in a phage P1-derived artificial chromosome (PAC) clone containing most of the human AE1 gene and the upstream flanking region. We determined its heterozygosity value in multiple populations by PCR analysis. Genotyping of one family with dominant dRTA identified the AE1 R589H mutation, and family member genotypes were compared with the CA repeat length. AE1 and vH(+)-ATPase polypeptides in kidney tissue from an AE1 R589H patient were examined by immunocytochemistry for the first time. RESULTS: This CA repeat, previously reported as D17S1183, is approximately 90 kb upstream of the AE1 gene and displayed considerable length polymorphism, with small racial differences, and a heterozygosity value of 0.56. The allele-specific length of this repeat confirmed co-segregation of the AE1 R589H mutation with the disease phenotype in a family with dominant dRTA. Immunostaining of the kidney cortex from one affected member with superimposed chronic pyelonephritis revealed vH(+)-ATPase-positive intercalated cells in which AE1 was undetectable, and proximal tubular epithelial cells with apparently enhanced apical vH(+)-ATPase staining. CONCLUSIONS: The highly polymorphic dinucleotide repeat adjacent to the human AE1 gene may be useful for future studies of disease association and haplotype analysis. Intercalated cells persist in the end-stage kidney of a patient with familial autosomal dominant dRTA associated with the AE1 R589H mutation. The absence of detectable AE1 polypeptide in those intercalated cells supports the genetic prediction that the AE1 R589H mutation indeed causes dominant dRTA. PMID- 14736962 TI - Expression of human nephrin mRNA in diabetic nephropathy. AB - BACKGROUND: Diabetic nephropathy (DN) is associated with functional changes in the filtration barrier, and microalbuminuria is a strong predictor of the development of overt DN. Nephrin is a novel podocyte-specific protein which localizes at the slit diaphragm. This study examines the expression of nephrin mRNA in the kidneys of type 2 diabetics with DN. METHODS: Renal tissues were obtained from 13 type 2 diabetics with DN. We also examined samples from five patients with minimal change nephrotic syndrome (MCNS) and five normal kidneys (normals) as control. The severity of DN was classified into two grades based on histopathological findings. DN grade 1 (DN1 = seven patients) presented mild mesangial expansion, and DN grade 2 (DN2 = six patients) moderate mesangial expansion. Nephrin mRNA was quantitated and localized by in situ hybridization. RESULTS: Cells positive for nephrin mRNA were detected exclusively in glomerular epithelial cells. The percentage of cells positive for nephrin mRNA in DN2 was significantly lower than in MCNS and normal kidneys. Furthermore, there was an inverse correlation between the percentage of cells positive for nephrin mRNA and extent of proteinuria. CONCLUSION: The low expression of nephrin mRNA may be closely linked to development and/or progression of proteinuria in human diabetic nephropathy. PMID- 14736963 TI - The renal tolerance of low-dose adefovir dipivoxil by lamivudine-resistant individuals co-infected with hepatitis B and HIV. AB - BACKGROUND: Adefovir (ADV), an orally administered nucleotide analogue active against hepadnaviruses, retroviruses and herpes viruses was shown to be effective in HIV-infected patients, but the prevalence of nephrotoxicity with doses of 60 120 mg/day was considered unacceptable. Recently, lower doses of ADV were shown to be effective for the treatment of HIV-1 patients with chronic lamivudine (LAM) resistant hepatitis B. METHODS: In a cohort of 35 patients infected with both HIV 1 and LAM-resistant hepatitis B virus, we investigated the renal tolerance of a once-daily dose of ADV 10 mg over 52 weeks. Their mean baseline creatinine clearance was within the normal range (105 +/- 3 ml/min/1.73 m(2)). No patient had significant changes in renal function or electrolyte balance secondary to ADV treatment. RESULTS: Transient increases in serum creatinine, which resolved by the end of the study were noted in two patients and three developed proteinuria, which was felt to be unrelated to ADV treatment. The cohort's mean serum phosphate level, 2.45 +/- 0.09 mg/dl at baseline, did not change significantly under treatment (2.66 +/- 0.12 mg/dl at week 52, P = NS). CONCLUSIONS: Our study shows that ADV dosed at 10 mg/day for the treatment of LAM-resistant chronic hepatitis B in patients co-infected with HIV is not associated with renal tubular dysfunction or a significant change in renal function. PMID- 14736964 TI - The efficacy and safety of B-type natriuretic peptide (nesiritide) in patients with renal insufficiency and acutely decompensated congestive heart failure. AB - BACKGROUND: Nesiritide (B-type natriuretic peptide) reduces preload and afterload, and causes natriuresis, diuresis and suppression of norepinephrine, endothelin-1 and aldosterone. In this study, we sought to explore the safety and efficacy of nesiritide in patients with acute congestive heart failure (CHF) and renal insufficiency (RI). METHODS: We studied the effects of nesiritide in patients with RI in the VMAC trial database, a multi-centre, randomized controlled trial (n = 489) of patients with acute decompensated CHF. RESULTS: The mean serum creatinine (SCr) in nesiritide-treated patients with RI (SCr > or = 2.0 mg/dl, n = 60, range 2.0-11.1 mg/dl) and without RI (SCr < 2.0 mg/dl, n = 209) was 3.0+/-1.51 and 1.2+/-0.34 mg/dl, respectively. Pulmonary capillary wedge pressure (PCWP) was reduced significantly and similarly in both RI and no RI groups starting at 15 min into nesiritide infusion from a baseline of 29.9+/-8.1 and 26.6+/-6.0 mmHg, respectively. Addition of placebo to standard therapies yielded no further improvement in PCWP in patients with RI; in contrast, nesiritide significantly reduced PCWP at every time point during 24 h. The effects of nitroglycerin were less robust than those of nesiritide, and PCWP was not significantly reduced by nitroglycerin at the 3 h primary end-point. At 24 h, 83% of the RI patients and 91% of patients without RI treated with nesiritide reported improvements in dyspnoea. Nesiritide was well tolerated in patients with RI and no RI, and renal function was preserved in both groups. CONCLUSIONS: In patients with RI, nesiritide was safe and improved haemodynamics and dyspnoea. PMID- 14736965 TI - Evaluation of an in vitro dialysis system to predict drug removal. AB - BACKGROUND: Variation in the extent of drug removal under different dialysis conditions presents a challenge for prediction of drug elimination and dosage regimen adjustment during haemodialysis (HD). Dependence on clinical pharmacokinetic studies in HD patients for dosing guidelines is problematic given the increasing number of dialysers with variable rates of drug removal. Thus, the purpose of this study was to characterize drug removal using an in vitro system and to evaluate its reliability to predict in vivo elimination by HD using vancomycin (VANC) as a model drug. METHODS: In vitro dialysis was performed for 2 h (volume 4.0 l normal saline, initial VANC concentration 30 mg/l, flow rate 300 ml/min, dialysate flow 800 ml/min) using four different dialysers: polymethylmethacrylate (BK-2.1 U), polysulfone (F-80), AN69 (Filtral-20) and hemophan (COBE 700HE). The in vitro dialysis clearance for VANC (CL(D)) for the polysulfone dialyser was compared with values determined in eight HD patients. In vitro VANC CL(D) for all dialysers was compared with the clearance and KoA for B12 reported for each dialyser. RESULTS: In vitro VANC CL(D) values were 93+/-11 ml/min for the polymethylmethacrylate BK-2.1, 136+/-7 ml/min for the AN69, 65+/-9 ml/min for the hemophan COBE 700HE and 143+/-10 ml/min for the polysulfone F80. The CL(D) for the polysulfone F80 was not statistically different from the in vivo CL(D) of 135+/-18 ml/min (P = 0.48). In vitro VANC CL(D) correlated with the B12 CL(D) (r(2) = 0.77) and the B12 KoA (r(2) = 0.63) reported for each dialyser. CONCLUSION: VANC CL(D) in HD patients for the polysulfone dialyser was correctly predicted using the in vitro dialysis system. Use of this system may be superior to estimations of drug CL(D) based on dialyser information provided by the manufacturer for compounds of similar molecular weight. PMID- 14736966 TI - Analysis of factors associated with variability in haemodialysis adequacy. AB - BACKGROUND: Delivered dialysis dose measured as Kt/V is widely accepted as an important factor affecting mortality on haemodialysis. It is currently measured relatively infrequently in most units for pragmatic reasons. We have previously shown that significant variation occurs within individuals which often makes the difference between dialysis that would be considered adequate, and inadequate. The aim of this study is to delineate which factors are responsible for the observed variation. METHODS: We studied 1109 treatments in 109 patients, mean age 59.9 years, mean dry weight 71.8 kg. Depurated volume (Kt) was measured by ionic dialysance for each treatment. Kt and other relevant treatment-related variables were automatically recorded on a central server. Multivariate analysis using mixed models with backwards elimination was used to analyse the determinants of Kt. Kt/V was not used, in order to avoid introducing error in the determination of V. RESULTS: The analysis indicated that the following were independent determinants of Kt: blood pump speed, time, minimum and mean arterial line pressure, both maximum and minimum venous line pressures and total ionic mass balance. CONCLUSION: This analysis suggests that the variation in adequacy that occurs within an individual is multifactorial. It confirms the importance of effective vascular access, prescription of and adherence to adequate time on dialysis, and reinforces the impact of degree of sodium removal. In clinical practice absolute control of these variables is not possible in every dialysis session and some degree of variability in Kt is therefore inevitable. Monitoring of adequacy thus requires more frequent assessment of Kt than is currently performed. Online monitoring of ionic dialysance achieves this. PMID- 14736967 TI - Predicting 1 year mortality in an outpatient haemodialysis population: a comparison of comorbidity instruments. AB - BACKGROUND: A valid and practical measure of comorbid illness burden in dialysis populations is greatly needed to enable unbiased comparisons of clinical outcomes. We compare the discriminatory accuracy of 1 year mortality predictions derived from four comorbidity instruments in a large representative US dialysis population. METHODS: Comorbidity information was collected using the Index of Coexistent Diseases (ICED) in 1779 haemodialysis patients of a national dialysis provider between 1997 and 2000. Comorbidity was also scored according to the Charlson Comorbidity Index (CCI), Wright-Khan and Davies indices. Relationships of instrument scores with 1 year mortality were assessed in separate logistic regression analyses. Discriminatory ability was compared using the area under the receiver-operating characteristics curve (AUC), based on predictions of each regression model. RESULTS: When mortality was predicted using comorbidity and age, the ICED better discriminated between survivors and those who died (AUC 0.72) as compared with the CCI (0.67), Wright-Khan (0.68) and Davies (0.68) indices. Upon addition of race and serum albumin, predictive accuracy of each model improved further (AUCs of the ICED, 0.77; CCI, 0.75; Wright-Khan Index, 0.75; Davies Index, 0.74). CONCLUSIONS: The ICED had greater discriminatory ability than the CCI, Davies and Wright-Khan indices, when age and a comorbidity index were used alone to predict 1 year mortality; however, the differences among instruments diminished once serum albumin, race and the cause of ESRD were accounted for. None of the currently available comorbidity instruments tested in this study discriminated mortality outcomes particularly well. Assessing comorbidity using the ICED takes significantly more time. Identifying the key prognostic comorbid conditions and weighting these according to outcomes in a dialysis population should increase accuracy and, with restriction to a finite number of items, provide a practical means for widespread comorbidity assessment. PMID- 14736968 TI - Associations between plasma ghrelin levels and body composition in end-stage renal disease: a longitudinal study. AB - BACKGROUND: Ghrelin is a newly detected orexigenic gastric hormone that stimulates food intake. Increased levels of ghrelin are often found in disease states associated with wasting. Wasting is a common phenomenon in end-stage renal disease (ESRD) patients in whom elevated ghrelin levels have been reported. However, no data are available on the relationship between body composition and plasma ghrelin levels in this patient group. METHODS: The study population consisted of 108 (71 males) ESRD patients aged 53+/-12 years. Body composition, nutritional status (subjective global assessment), estimated protein intake (nPNA), plasma ghrelin, plasma insulin and serum leptin were evaluated close to the start of dialysis treatment. Twelve healthy subjects (nine males, 44+/-6 years) served as the control group. A longitudinal evaluation of changes in plasma ghrelin and body composition was performed in 52 of the patients after 12 months of dialysis treatment. RESULTS: Markedly elevated plasma ghrelin levels (843+/-485 vs 443+/-302 pg/ml; P<0.01) were observed in ESRD patients compared with controls. Basal plasma ghrelin levels correlated significantly with plasma insulin (R = -0.32; P<0.05), body mass index (R = -0.24; P<0.05), log serum leptin levels (R = -0.23; P<0.05) and truncal fat mass (R = -0.25; P<0.05). The longitudinal analysis of body composition demonstrated that whereas fat mass increased (23.7+/-8.6 to 25.3+/-9.9 kg; P<0.05) and plasma ghrelin levels decreased (855+/-429 to 693+/-408 pg/ml; P<0.05) significantly in peritoneal dialysis patients, no significant changes in either body composition or plasma ghrelin levels were found in patients treated by haemodialysis. CONCLUSION: Markedly elevated plasma ghrelin levels are found in advanced renal failure and correlate with fat mass, plasma insulin and serum leptin levels. Changes in plasma ghrelin during 12 months of peritoneal dialysis treatment are associated with changes in body composition. PMID- 14736969 TI - Underweight rather than overweight is associated with higher prevalence of hypertension: BP vs BMI in haemodialysis population. AB - BACKGROUND: Recent surveys have reported a higher prevalence of hypertension and obesity in the haemodialysis population, suggesting a possible link between these two parameters. Alternatively, malnutrition through proinflammatory and antiendothelial mechanisms may contribute to hypertension in malnourished, underweight patients on haemodialysis, whereas the converse may be true in overweight patients. METHODS: The relationship between blood pressure (BP) and body mass index (BMI) was examined in 1010 patients on chronic haemodialysis using univariate and multivariate analyses. RESULTS: Based on a systolic BP of >150 mmHg, 38% were hypertensive in the post-dialysis period. A similar figure for a diastolic BP of >90 mmHg was 21%. Unlike in the general population, there was no positive correlation between BP and BMI in simple regression analysis (r = -0.13, P = 0.0005). The percentage of patients with systolic hypertension was higher among the underweight (BMI <20) than the normal weight (BMI 20-27.5) or overweight patients (BMI >27.5) and were 35, 29 and 20%, respectively. Furthermore, a negative correlation, albeit weak, existed between systolic BP and markers of nutrition such as serum prealbumin (r = -0.10, P = 0.03) and serum creatinine (r = -0.12, P = 0.008), suggesting a plausible link between impaired nutrition and elevated BP. In multiple regression analysis, the lack of a positive correlation between BP and BMI persisted (r = -0.16, P = 0.0007) in spite of adjusting for case-mix characteristics and parameters related to BP and nutrition. CONCLUSIONS: We demonstrate for the first time that, unlike the general population, no positive correlation exists between BP and increasing BMI in haemodialysis patients. Further analysis is necessary to verify our observation and to test the possibility of whether nutritional improvement would aid in the better control of hypertension in dialysis patients. PMID- 14736970 TI - The value of tuberculin skin testing in haemodialysis patients. AB - BACKGROUND: Chronic haemodialysis patients are at increased risk for developing tuberculosis (TB). Appropriate screening methods to detect latent Mycobacterium tuberculosis infection are required. The aim of this prospective multi-centre study was to evaluate the tuberculin skin test (TST) as a screening method for detection of M.tuberculosis infection in haemodialysis patients. METHODS: A total of 224 patients in two haemodialysis centres were prospectively tested, using 2 units of tuberculin PPD RT23. Up to three booster injections were given with a 7 day interval to patients not responding to the previous test. The results were compared with clinical and radiological data. RESULTS: The cumulative prevalence of a positive TST was 14.7% for the first test, 27.8% for the second test and 32.6% for the fourth test. There was no influence of age, gender, haemodialysis centre, dialysis efficiency, nutritional state, levels of zinc, vitamin D therapy, primary renal disease, (previous or active) immunosuppressive therapy or response to hepatitis B vaccination. There was a significant, but weak, correlation between TST positivity and a history of positive TST or TB. Chest radiography and positive TST were not correlated, yet a positive chest X-ray increased the detection of patients with latent M.tuberculosis infection up to 47.8%. CONCLUSIONS: In haemodialysis patients, a positive response of >30% to repeated TST was obtained. Two consecutive TSTs were sufficient to recruit most of the booster reactions. Since only a weak correlation was found with anamnestic data, regular TST evaluation in combination with a chest X-ray, is a useful tool to detect infection with M.tuberculosis in haemodialysis patients. PMID- 14736971 TI - Association of donor TNFRSF6 (FAS) gene polymorphism with acute rejection in renal transplant patients: a case-control study. AB - BACKGROUND: Genetic factors other than HLA have been reported to be associated with the outcome of organ transplantations. Because binding of FasL to its receptor Fas could play an important role in tubulitis and in the death of graft tubular epithelial cells during kidney allograft rejection, a gene polymorphism recently identified in position -671 in the promoter of the TNFRSF6 gene coding for Fas was investigated in donors. METHODS: A case-control study was performed within a cohort of non-hyperimmunized adult patients who had received cadaveric kidney transplants based on the occurrence or absence of acute cellular rejection in the first 6 months after renal transplantation. Each recipient from the acute rejection group (n = 35) was matched for age (+/- 5 years) and number of HLA-DR mismatches with two recipients within the non-acute rejection group (n = 70). RESULTS: The TNFRSF6-GG genotype was more frequent in donors in the group without rejection episodes. In contrast, patients who received a kidney from a TNFRSF6-A carrier were more likely to experience acute rejection episodes (relative risk nearly 2.1). CONCLUSION: This study suggests that donor TNFRSF6 polymorphism directly or indirectly influences acute kidney rejection episodes. PMID- 14736972 TI - Hyponatraemia and hyperkalaemia are more frequent in renal transplant recipients treated with tacrolimus than with cyclosporin. Further evidence for differences between cyclosporin and tacrolimus nephrotoxicities. AB - BACKGROUND: This study was designed to examine the hypothesis that the nephrotoxicities caused by cyclosporin and tacrolimus might differ in respect of sodium and potassium handling. METHODS: 125 patients were studied retrospectively for the first 90 days after renal transplantation. Eighty were treated initially with cyclosporin and 45 with tacrolimus. RESULTS: A serum sodium level of <135 mmol/l was present for 542/5171 (10.5%) days under tacrolimus treatment compared with 377/5486 (6.9%) days under cyclosporin treatment (P < 0.0001). Severe hyponatraemia, below 120 mmol/l, was also more prevalent under tacrolimus than cyclosporin treatment, P < 0.025. Nine patients, all receiving tacrolimus, were treated with fludrocortisone for fluid depletion and/or hyponatraemia. Serum potassium levels were higher in tacrolimus-treated patients (P < 0.0001), and subjects with hyponatraemia were more likely to experience hyperkalaemia (P < 0.0001). CONCLUSIONS: Hyponatraemia and hyperkalaemia were more frequent in tacrolimus-treated subjects. Taken together with previous work showing that hyperuricaemia is more frequent with cyclosporin treatment, and hypomagnesaemia with tacrolimus treatment, these findings are consistent with qualitative differences between the nephrotoxicities of cyclosporin and tacrolimus. PMID- 14736973 TI - Mercury exposure in protein A immunoadsorption. AB - BACKGROUND: Immunoadsorption is increasingly used to treat antibody-mediated autoimmune diseases. To prevent microbial growth during storage, reusable protein A-Sepharose gel columns are primed with ethyl mercury thiosalicylate (thiomersal, 0.1% solution) and rinsed with phosphate buffer before use. In this study, we tested the hypothesis of systemic mercury exposure in protein A immunoadsorption. METHODS: Whole blood mercury levels were measured by atomic absorption spectroscopy before and after protein A immunoadsorption (11 patients, 26 treatments), anti-IgG immunoadsorption (eight patients, 13 treatments) and LDL apheresis (DALI and Therasorb systems; nine patients, 14 treatments). RESULTS: Patients treated with protein A immunoadsorption had significantly elevated baseline mercury levels compared with the other groups, which were not different from healthy controls. Following protein A immunoadsorption, mercury levels increased from 5.9+/-1.4 microg/l (mean+/-SEM, normal, <5 microg/l) to 32.3+/-5.7 microg/l, P<0.001). In one intensively treated patient, acute neurological toxicity developed at a mercury level of 107 microg/l. Symptoms abated slowly and did not recur after switching to a thiomersal-free system and chelation therapy. No mercury release to patients occurred in anti-IgG immunoadsorption or LDL apheresis treatments. CONCLUSION: This preliminary report suggests that protein A immunoadsorption columns primed with thiomersal during storage may cause a sustained increase of systemic mercury concentrations, which exceed current safety recommendations in a proportion of patients. Considering the potential for mercury-induced toxicity, every effort should be undertaken to reduce systemic mercury exposure, either by adding chelators to the rinsing solution or ideally by replacement of thiomersal. PMID- 14736975 TI - Automated flow cytometry analysis of peritoneal dialysis fluid. AB - BACKGROUND: Recently, the Sysmex UF-100 flow cytometer has been developed to automate urinalysis. We have evaluated this instrument to explore the possibilities of flow cytometry in the analysis of peritoneal dialysis fluid (PD) and have compared the obtained data with those of counting chamber techniques, biochemical analysis and bacterial culture. METHODS: UF-100 data were correlated with microscopy and biochemical data in 135 PD samples. Microbiological analysis was performed in 63 suspected cases of peritonitis. RESULTS: Good agreement (P < 0.001) was obtained between UF-100 and microscopy data for leukocytes (r = 0.825). UF-100 bacterial count correlated (P < 0.001) with UF-100 leukocyte count (r = 0.549). UF-100 bacterial counts were unreliable in samples where interference by blood platelets was observed. Another major problem was the UF 100 'bacterial' background signal in sterile PD samples. Yeast cells were detected by the flow cytometer in spiked samples. CONCLUSIONS: Flow cytometry of PD with the UF-100 offers a rapid and reliable leukocyte count. Sensitivity of the 'bacterial' channel count in predicting positive culture exceeds the sensitivity of conventional Gram stain. Furthermore, additional semi-quantitative information is provided regarding the presence of yeasts. PMID- 14736974 TI - Serum uric acid levels show a 'J-shaped' association with all-cause mortality in haemodialysis patients. AB - BACKGROUND: Although elevated serum levels of uric acid are common in patients with kidney disease or in those receiving maintenance dialysis therapy, the clinical impact of uric acid on mortality in haemodialysis (HD) patients remains unclear. This work was designed to explore the predictive value of serum uric acid levels on all-cause mortality of HD patients. METHODS: We retrospectively analysed mortality rates in 146 chronic HD patients that were treated with HD three times per week at our HD unit for a period of one full year. The analysed parameters included demographic characteristics, aetiology of end-stage renal disease, co-morbid conditions, duration (at least 1 year) and delivered dose of HD, normalized protein catabolic rate, serum albumin concentration, haematocrit, serum uric acid (UA) levels and other laboratory parameters. A multivariate Cox proportional hazards model, which included adjustment for the above factors, was applied to identify the predictive value of UA levels on patient mortality. RESULTS: A Cox proportional hazards model revealed that decreased serum albumin, underlying diabetic nephropathy (DMN) and UA groups (< or =20th, 20-80th and > or =80th percentiles; P = 0.016) were all significant, independent predictors of all cause mortality in HD patients. The hazard ratios of death were: serum albumin (per 0.5 g/dl decrease), 3.10 [95% confidence interval (95% CI), 1.80-5.34, P < 0.001]; DMN (vs non-DMN), 3.47 (95% CI, 1.25-9.59, P = 0.017); and UA groups (vs 20th to 80th percentile): < or =20th percentile, 2.98 (95% CI, 0.82-10.90, P = 0.099); > or = 80th percentile, 5.67 (95% CI, 1.71-18.78, P = 0.004). CONCLUSIONS: These preliminary observations suggest that HD patients in the lowest and highest quintiles of UA levels would face higher risk of mortality. Further studies with larger sample sizes will be needed to confirm these findings. PMID- 14736976 TI - Nephrogenic fibrosing dermopathy: a novel, disabling disorder in patients with renal failure. PMID- 14736977 TI - IgG4-associated idiopathic tubulointerstitial nephritis complicating autoimmune pancreatitis. PMID- 14736978 TI - Nephrotic syndrome: cause of an abnormal response to the rapid ACTH stimulation test. PMID- 14736979 TI - Low-dose cyclosporin therapy for recombinant erythropoietin-induced pure red-cell aplasia. PMID- 14736980 TI - Nephrotic range proteinuria in a renal transplant associated with oncocytoma of the native kidney. PMID- 14736981 TI - Granulomatous interstitial nephritis of the allograft kidney associated with rhodococcal pulmonary infection. PMID- 14736982 TI - Recurrence of anti-GBM disease 8 years after renal transplantation. PMID- 14736983 TI - Renal artery dissection secondary to medial hyperplasia presenting as loin pain haematuria syndrome. PMID- 14736984 TI - Renal functional assessment in the failing renal graft: what to do where clearances show discrepancy. PMID- 14736985 TI - Abdominal gas is not always bowel associated: lessons from an allograft recipient. PMID- 14736986 TI - Uraemic tumoural calcinosis. PMID- 14736989 TI - Albumin and glomerular permselectivity. PMID- 14736990 TI - Renal infarction due to combination of fibromuscular dysplasia and factor V Leiden mutation. PMID- 14736991 TI - Acute intratubular obstructive renal failure after ampicillin treatment. PMID- 14736992 TI - Measles susceptibility in haemodialysis patients in Argentina. PMID- 14736993 TI - Drug interaction between sevelamer and cyclosporin. PMID- 14736994 TI - Use of the QTc interval and QTc dispersion in patients on haemodialysis: assessment of reproducibility. PMID- 14736996 TI - The major 5' determinant in stop codon read-through involves two adjacent adenines. AB - The aim of this approach was to identify the major determinants, located at the 5' end of the stop codon, that modulate translational read-through in Saccharomyces cerevisiae. We developed a library of oligonucleotides degenerate at the six positions immediately upstream of the termination codon, cloned in the ADE2 reporter gene. Variations at these positions modulated translational read through efficiency approximately 16-fold. The major effect was imposed by the two nucleotides immediately upstream of the stop codon. We showed that this effect was neither mediated by the last amino acid residues present in the polypeptide chain nor by the tRNA present in the ribosomal P site. We propose that the mRNA structure, depending on the nucleotides in the P site, is the main 5' determinant of read-through efficiency. PMID- 14736997 TI - Interactive effects of vinclozolin and testosterone propionate on pregnancy and sexual differentiation of the male and female SD rat. AB - In mammals, androgens are essential in directing mammalian sexual differentiation of the male phenotype. Administration of testosterone during this period alters female development in a male-like direction, whereas exposure to an androgen receptor antagonist like vinclozolin (V) demasculinizes and feminizes the male offspring. In the current study, we administered V (gavage at 200 mg/kg/day) and/or testosterone propionate (TP, sc, at 1 mg/rat/day), alone and in combination to Sprague-Dawley (SD) rats on days 14 through 19 of pregnancy, to determine if V would antagonize the effects of TP in the female and, conversely, if TP would antagonize the effects of V in the male offspring. These doses of TP and V were selected because they significantly alter sexual differentiation in the majority of female and male rat offspring, respectively, without producing severe toxicity in the dam or offspring. The study design is a 2 x 2 factorial (7 dams per group) including vehicle control, V, TP, and V + TP groups. As expected, individually, both V and TP reduced maternal weight gain and the V + TP group was affected in a cumulative fashion. Litter size on postnatal day (PND) 2 was reduced only by V + TP, whereas pup body weight was reduced in all three treated groups, the effect of V + TP again being cumulative. In female offspring, TP induced alterations (i.e., increased anogenital distance [AGD] and fewer nipples, vaginal agenesis, hydrometrocolpos, induced prostate and bulbourethral glands, and levator ani muscle tissues) were all reversed by coadministration of V. In male offspring, V-induced alterations were only modestly antagonized by TP. At the dosage levels used herein, V + TP-treated male offspring had less well developed nipples as infants and adults and a lower incidence of ectopic testis than did the V group. However, V-induced changes in reproductive organ weights, AGD, atrophic testes, vaginal pouch, and agenesis of the sex accessory tissues were not antagonized by concurrent TP treatment in male offspring. We observed that the combination of V and TP, two chemicals with opposing endocrine action, antagonized one another during sexual differentiation, especially in the female offspring and induced cumulative effects on maternal and neonatal toxicity. We suspect that antagonism of V by TP would be enhanced in the male if lower dose levels of V were used, but then the antagonism of TP by V in the female would likely be attenuated. PMID- 14736998 TI - Different mechanisms mediate uptake of lead in a rat astroglial cell line. AB - The mechanism by which lead (Pb) enters astrocytes was examined in a rat astroglial cell line in order to characterize specific pathways for transport. Pb uptake was saturable at pH 5.5 and 7.4, although quantitative differences existed in the Michaelis-Menten constants. At pH 7.4, the Vmax and Km were 2700 fmoles/mg protein/min and 13.4 microM, respectively, whereas the Vmax and Km were 329 fmoles/mg and 8.2 microM in the buffer at pH 5.5, respectively. The presence of extracellular iron inhibited uptake in a buffer at pH 5.5 but not at pH 7.4. Cells treated with the iron chelator deferoxamine displayed higher levels of the iron transporter divalent metal transporter 1 (DMT1) mRNA and protein, and consistent with increased DMT1 expression, the treated cells displayed greater uptake of Pb in the buffer at pH 5.5 but not at pH 7.4. Alternatively, at pH 7.4, the transport of Pb was blocked by the anion transporter inhibitor 4,4' diisothiocyanatodihydrostilbene-2,2'-disulfonic acid (DIDS), which bound to cell surface proteins at concentrations that were similar to those that blocked Pb uptake. DIDS did not inhibit uptake of Pb in the buffer at pH 5.5. Greater uptake of Pb was observed in a buffer containing sodium bicarbonate, which was abrogated in the presence of DIDS. In summary, the astroglial cell line displays two distinct pH-sensitive transport mechanisms for Pb. PMID- 14736999 TI - 2,3,7,8-Tetrachlorodibenzo-p-dioxin inhibits regression of the common cardinal vein in developing zebrafish. AB - A role for the aryl hydrocarbon receptor (AHR) pathway in vascular maturation has been implicated by studies in Ahr-null mice. In this study the hypothesis that activation of AHR signaling by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) alters common cardinal vein (CCV) development in the zebrafish embryo was investigated. The CCV is a paired vessel that grows across the yolk, connecting to the heart. It is extensively remodeled and regresses as the heart migrates dorsally within the pericardium. TCDD significantly reduced CCV growth as early as 44 h post fertilization (hpf), and CCV area was reduced to 63% of control at 62 hpf. This vascular response to TCDD was at least as sensitive as previously defined endpoints of TCDD developmental toxicity in zebrafish. TCDD also blocked regression of the CCV (by 80 hpf), possibly contributing to the "string-like" heart phenotype seen in TCDD-exposed zebrafish larvae. Dependence of the block in CCV regression on zebrafish (zf) AHR2 was investigated using a zfahr2 specific morpholino to knock down expression of AHR2. The zfahr2 morpholino had no effect on CCV regression in the absence of TCDD, but did protect against the TCDD induced block of CCV regression. This demonstrates that the TCDD-induced block in CCV regression is AHR2 dependent. It is significant that decreased CCV growth occurs before and inhibition of CCV regression occurs concurrent with overt signs of TCDD developmental toxicity. This suggests that alterations of vascular growth and remodeling may play a role in TCDD developmental toxicity in zebrafish. PMID- 14737000 TI - Epoxide hydrolases in the rat epididymis: possible roles in xenobiotic and endogenous fatty acid metabolism. AB - Epoxide hydrolases play an important role in detoxifying epoxides that arise from the metabolism of xenobiotic and endogenous compounds. Both the soluble and microsomal forms of epoxide hydrolase (sEH and mEH, respectively) have been detected in the rat testis. Because of the important role the epididymis plays in sperm maturation and protection, the present study evaluated the presence and activity of these two epoxide hydrolases in the rat epididymis. Using Western blotting, protein bands consistent in size with both mEH and sEH were detected in the caput, corpus, and cauda of the epididymis. The mEH immunoreactive bands in the epididymis ( approximately 50 kDa) were consistent with mEH detected in the liver and kidney. The sEH immunoreactive bands in the epididymis ( approximately 65 kDa) were consistent with a recombinant sEH standard and sEH detected in the liver, kidney, and testis. The presence of mEH and sEH in the epididymis was supported by observations from substrate-based enzyme assays. Results indicated that epididymal mEH can hydrolyze [(3)H]-cis-stilbene oxide to the corresponding diol at levels approximately 9% of the kidney. Epididymal sEH hydrolyzed the substrate [(3)H]-trans-diphenylpropene oxide to the corresponding diol and this activity was inhibited by cyclohexyl-dodecyl urea. Arachidonic acid epoxygenase activity was detected in epididymal S9 fractions, suggesting that fatty acid metabolism by epididymal cytochrome P450s can form epoxides that subsequently become substrates for epididymal sEH. Results from the present study indicate that the epididymis contains at least two active forms of epoxide hydrolase. The role of these enzymes in the detoxification of xenobiotic epoxides is well known, although it is unclear what cellular role they may play in the formation of biologically active metabolites in the epididymis. PMID- 14737001 TI - In vitro toxicity of kava alkaloid, pipermethystine, in HepG2 cells compared to kavalactones. AB - Kava herbal supplements have been recently associated with acute hepatotoxicity, leading to the ban of kava products in approximately a dozen countries around the world. It is suspected that some alkaloids from aerial kava may have contributed to the problem. Traditionally, Pacific Islanders use primarily the underground parts of the shrub to prepare the kava beverage. However, some kava herbal supplements may contain ingredients from aerial stem peelings. The aim of this study was to test the in vitro effects of a major kava alkaloid, pipermethystine (PM), found mostly in leaves and stem peelings, and kavalactones such as 7,8 dihydromethysticin (DHM) and desmethoxyyangonin (DMY), which are abundant in the roots. Exposure of human hepatoma cells, HepG2, to 100 microM PM caused 90% loss in cell viability within 24 h, while 50 microM caused 65% cell death. Similar concentrations of kavalactones did not affect cell viability for up to 8 days of treatment. Mechanistic studies indicate that, in contrast to kavalactones, PM significantly decreased cellular ATP levels, mitochondrial membrane potential, and induced apoptosis as measured by the release of caspase-3 after 24 h of treatment. These observations suggest that PM, rather than kavalactones, is capable of causing cell death, probably in part by disrupting mitochondrial function. Thus, PM may contribute to rare but severe hepatotoxic reactions to kava. PMID- 14737002 TI - 2,2',4,6,6'-Pentachlorobiphenyl induces mitotic arrest and p53 activation. AB - Polychlorinated biphenyls (PCBs), a class of persistent organic pollutants (POPs), have been considered to be involved in cancers, but the underlying mechanisms are not known well. Various cancers are closely related to genetic alteration; therefore, we investigated the effect of PCBs on genetic stability, through p53, a guardian of genome, in NIH 3T3 fibroblasts. Among several congeners examined, 2,2',4,6,6'-pentachlorobiphenyl (PeCB) specifically activated p53-dependent transcription. It also induced p53 nuclear accumulation, but did not cause DNA strand breakage. On the other hand, cell cycle progression that is closely connected to p53 was affected by 2,2',4,6,6'-PeCB, resulting in mitotic arrest. In the arrested cells, mitotic spindle damage was detected. Moreover, in the absence of functional p53, polyploidy was caused by 2,2',4,6,6'-PeCB. These results imply that 2,2',4,6,6'-PeCB induces mitotic arrest by interfering with mitotic spindle assembly, followed by genetic instability which triggers p53 activating signals to prevent further polyploidization. Taking these findings together, we suggest that 2,2',4,6,6'-PeCB could be involved in cancer development by causing genetic instability through mitotic spindle damage, which brings about aneuploidy in p53-deficient tumor cells. PMID- 14737003 TI - Motorcycle exhaust particles induce airway inflammation and airway hyperresponsiveness in BALB/C mice. AB - A number of large studies have reported that environmental pollutants from fossil fuel combustion can cause deleterious effects to the immune system, resulting in an allergic reaction leading to respiratory tract damage. In this study, we investigated the effect of motorcycle exhaust particles (MEP), a major pollutant in the Taiwan urban area, on airway inflammation and airway hyperresponsiveness in laboratory animals. BALB/c mice were instilled intratracheally (i.t.) with 1.2 mg/kg and 12 mg/kg of MEP, which was collected from two-stroke motorcycle engines. The mice were exposed 3 times i.t. with MEP, and various parameters for airway inflammation and hyperresponsiveness were sequentially analyzed. We found that MEP would induce airway and pulmonary inflammation characterized by infiltration of eosinophils, neutrophils, lymphocytes, and macrophages in bronchoalveolar lavage fluid (BALF) and inflammatory cell infiltration in lung. In addition, MEP treatment enhanced BALF interleukin-4 (IL-4), IL-5, and interferon-gamma (IFN-gamma) cytokine levels and serum IgE production. Bronchial response measured by unrestrained plethysmography with methacholine challenge showed that MEP treatment induced airway hyperresponsiveness (AHR) in BALB/c mice. The chemical components in MEP were further fractionated with organic solvents, and we found that the benzene-extracted fraction exerts a similar biological effect as seen with MEP, including airway inflammation, increased BALF IL-4, serum IgE production, and induction of AHR. In conclusion, we present evidence showing that the filter-trapped particles emitted from the unleaded gasoline-fueled two-stroke motorcycle engine may induce proinflammatory and proallergic response profiles in the absence of exposure to allergen. PMID- 14737004 TI - Sodium selenite-induced apoptosis in murine B-lymphoma cells is associated with inhibition of protein kinase C-delta, nuclear factor kappaB, and inhibitor of apoptosis protein. AB - Selenium (Se) is an essential trace element possessing anticarcinogenic properties and other biological functions. This study determined the role sodium selenite plays on intracellular signaling, including protein kinase C (PKC), nuclear factor-kappa B (NF-kappaB), and inhibitor of apoptosis protein (IAP) in murine B lymphoma (A20) cells. In vitro supplementation of A20 cells with low concentrations of sodium selenite (0.005-5 microM) caused a significant increase in cellular proliferation exclusively at 72 h. Proliferation and cell viability were decreased in response to selenium concentrations of >/= 25 microM and >/= 5 microM at 72 and 96 h, respectively. Flow cytometric analysis of A20 cells exposed to 5 microM Se at 72 and 96 h indicated G(2)-M phase arrest and increased cell death at higher concentrations. Se-induced cytotoxicity was associated with apoptosis indicated by nuclear fragmentation and DNA laddering. Se concentrations, which induced cell cycle arrest and apoptosis, were associated with inhibition of cytosol to membrane translocation of PKCdelta and PKC activity at 72 h. Coincubation of cultures with 0.5 microM phorbol 12-myristate 13-acetate (PMA) and Se (5 and 25 microM) reversed the Se-induced cell death at 72 h. The nuclear NF-kappaB translocation and NF-kappaB DNA-binding were inhibited by increasing concentrations of Se (5 and 25 microM) at 72 h. After 72 h exposure to 5 and 25 microM Se, cIAP-2 concentration was decreased. Differential inhibition of PKCdelta, NF-kappaB, and cIAP-2 by Se may represent important intracellular signaling processes through which Se induces apoptosis and subsequently exerts its anticarcinogenic potential. PMID- 14737005 TI - Evaluating the human relevance of chemically induced animal tumors. AB - Defining the mode(s) of action by which chemicals induce tumors in laboratory animals has become a key to judgments about the relevance of such tumor data for human risk assessment. Frameworks for analyzing mode of action information appear in recent U.S. EPA and IPCS publications relating to cancer risk assessment. This FORUM paper emphasizes that mode of action analytical frameworks depend on both qualitative and quantitative evaluations of relevant data and information: (1) presenting key events in the animal mode of action, (2) developing a "concordance" table for side-by-side comparison of key events as defined in animal studies with comparable information from human systems, and (3) using data and information from mode of action analyses, as well as information on relative sensitivity and exposure, to make weight-of-evidence judgments about the relevance of animal tumors for human cancer assessments. The paper features a systematic analysis for using mode of action information from animal and human studies, based in part on case examples involving environmental chemicals and pharmaceuticals. PMID- 14737006 TI - Platelets and anti-platelet therapy. AB - Platelets play a central role in the hemostatic process and consequently are similarly involved in the pathological counterpart, thrombosis. They adhere to various subendothelial proteins, exposed either by injury or disease, and subsequently become activated by the thrombogenic surface or locally produced agonists. These activated platelets aggregate to form a platelet plug, release agonists which recruit more platelets to the growing thrombus, and provide a catalytic surface for thrombin generation and fibrin formation. These platelet rich thrombi are responsible for the acute occlusion of stenotic vessels and ischemic injury to heart and brain. A range of anti-platelet drugs are currently used, both prophylactically and therapeutically, in regimens to manage thrombo embolic disorders. These include inhibitors of the generation, or effects, of locally produced agonists; several large clinical trials have supported roles for cyclooxygenase inhibitors, which prevent thromboxane generation, and thienopyridine derivatives, which antagonize ADP receptors. Similarly intravenous alpha IIb beta 3 antagonists have been shown to be effective anti-thrombotics, albeit in highly selective situations; in contrast, to date studies with their oral counterparts have been disappointing. Recent advances in understanding of platelet physiology have suggested several novel, if yet untested, targets for anti-platelet therapy. These include the thrombin receptor, the serotonin handling system, and the leptin receptor. PMID- 14737008 TI - New methods to evaluate endothelial function: method for assessing endothelial function in humans using a strain-gauge plethysmography: nitric oxide-dependent and -independent vasodilation. AB - The vascular endothelium is involved in the release of various vasodilators, including nitric oxide (NO), prostaglandins, and endothelium-derived hyperpolarizing factor as well as vasoconstrictors. NO plays an important role in the regulation of vascular tone, the inhibition of platelet aggregation, and the suppression of smooth muscle cell proliferation. Several diseases are associated with changes in endothelial function mediated through reduced NO bioavailability. In addition, endothelial dysfunction is an early feature of atherosclerosis and vascular diseases in humans. Therefore, it is clinically important to estimate the degree of endothelial dysfunction. Several methods have been used to assess endothelial function in humans. Recently, we have evaluated the effects of intra arterial infusion of infusion of NO agonists, such as acetylcholine, methacholine, and bradykinin, and NO antagonists on forearm blood flow using mercury-filled Silastic strain-gauge plethysmography. The response to the intra arterial infusion of vasoactive agents should be considered the gold standard in assessing endothelial function, because the use of agonists to stimulate NO release allow us to draw more specific conclusions concerning the role of basal and stimulated NO release. However, the invasive method is time-consuming and is a burden for patients. A noninvasive method of measuring forearm blood flow response to reactive hyperemia also is useful in assessing endothelial function. In this review we would like to explain in detail the methods of assessing endothelial function in humans using strain-gauge plethysmography. PMID- 14737009 TI - New methods to evaluate endothelial function: Non-invasive method of evaluating endothelial function in humans. AB - Evidence has accumulated that impairment of vascular endothelial function is the initial step in the development of atherosclerosis. One important finding is impairment of the release of endothelium-dependent relaxing factor, which is now thought to be nitric oxide or its related substances, from endothelial cells. Flow-mediated dilatation has been known to be endothelium-dependent, and this can be detected during reactive hyperemia by high-resolution ultrasound in superficial arteries. Several coronary risk factors have been reported to be significantly related with decreased flow-mediated dilatation. We studied the association between the accumulation of coronary risk factors (hyperlipidemia, diabetes mellitus, hypertension, and current smoking habitus) and vascular endothelial function. The lower incidence of atherosclerosis in women before the menopause than that in men is an established epidemiological observation. Short term estrogen therapy improves endothelial function in postmenopausal women. However, there are few reports on its long-term effects on endothelial function. Furthermore, we determined whether a reduced dosage of estrogen may maintain its beneficial effects. A similar improvement was also observed while women were on hormone replacement therapy even at the reduced dosage. Our results indicate that even at half the dose of estrogen, hormone replacement therapy may improve endothelial function in postmenopausal women. PMID- 14737010 TI - New methods to evaluate endothelial function: A search for a marker of nitric oxide (NO) in vivo: re-evaluation of NOx in plasma and red blood cells and a trial to detect nitrosothiols. AB - Although plasma NOx (NO(2)(-) and NO(3)(-)) has been used as an index of nitric oxide (NO) formation in vivo, many unreasonable results appeared even after active elimination of NOx contamination from laboratory ware. For example, plasma NOx concentrations did not increase during vasodilation mediated by the NO/cGMP pathway or after organ perfusion. A possible shift of NOx from plasma to erythrocytes (RBCs) as a cause of these phenomena has been excluded, leaving the destination of NOx (after leaving plasma) unknown. Kinetic analyses have revealed that steady state NOx concentrations in plasma and whole blood did not correlate with the NOx formation rate, but rather with the NOx elimination rate. Therefore, the supposition that the NO status is directly reflected by plasma NOx concentrations appears untenable. As nitrosothiols (R-SNOs), possible carriers of NO bioactivity, have been flagged as alternative indices of NO status in vivo, efforts have been made to detect these substances. When interference by ultrafiltration was eliminated, low molecular weight R-SNOs such as nitrosocystein and nitrosogluthathione were undetectable. However, a high molecular weight R-SNO, nitrosoalbumin, was detected in human blood. Further research is required into the significance and practical use of nitrosoalbumin as a marker of NO in vivo. PMID- 14737011 TI - New methods to evaluate endothelial function: Evaluation of endothelial function by hemoglobin-nitric oxide complex using electron paramagnetic resonance spectroscopy. AB - This minireview describes the practical use of assay systems to detect nitric oxide (NO) by electron paramagnetic resonance (EPR) spectroscopy for evaluation of endothelial functions. The iron(II)-dithiocarbamate complexes, such as iron(II)-(N-methyl-D-glucamine dithiocarbamate), are commonly used in EPR detection of NO both in vivo and in vitro. However, due to their redox activity, these complexes have some drawbacks that limit their usefulness for the detection of NO. On the other hand, the measurement of hemoglobin-NO adduct (HbNO) in whole blood by the EPR method seems relevant for the assessment of systemic NO levels. However, ceruloplasmin and an unknown radical species overlapping the same magnetic field as that of HbNO, which makes it physically impossible to measure small amounts of HbNO. Thus, to reveal the EPR spectrum of HbNO, we developed the EPR signal subtraction method, which is based on the computer-assisted subtraction of the digitized EPR spectrum of HbNO-depleted blood from that of the sample blood using software. Using this technique, we succeeded in measuring the steady blood HbNO level as an index of NO by the EPR HbNO signal subtraction method. We also demonstrated that temocapril reduces abnormalities of NO dynamics in the L-NAME (N(omega)-nitro-L-arginine-methylester)-induced endothelial dysfunction of rats using the EPR HbNO signal subtraction method. PMID- 14737012 TI - Lack of mu-opioid receptor-mediated G-protein activation in the spinal cord of mice lacking Exon 1 or Exons 2 and 3 of the MOR-1 gene. AB - The G-protein activation induced by mu-opioid receptor agonists was determined in spinal cord membranes from two types of mu-opioid receptor knockout mice: mice with a disruption of exon 1 (MOR (Exon 1)-KO) or exons 2 and 3 (MOR (Exons 2 and 3)-KO) of the mu-opioid receptor gene. The G-protein activation induced by the opioid agonists was measured by monitoring the increases of guanosine-5'-O-(3 [(35)S]thio)triphosphate ([(35)S]GTP gamma S) binding. The mu-opioid receptor agonists (D-Ala(2),N-MePhe (4),Gly-ol(5)]enkephalin, endomorphin-1, endomorphin 2, morphine, morphine-6 beta-glucuronide, and fentanyl produced concentration dependent increases of [(35)S]GTP gamma S binding to spinal cord membranes in wild-type mice, but not in MOR (Exon 1)-KO mice or MOR (Exons 2 and 3)-KO mice. On the other hand, the delta-opioid receptor agonist [D-Pen (2,5)]enkephalin, the kappa-opioid receptor agonist (-)U50,488H, or the ORL1-receptor agonist nociception increased [(35)S]GTP gamma S binding in the spinal cord membranes from both MOR (Exon 1)-KO mice and MOR (Exons 2 and 3)-KO mice to the same extent as in the corresponding wild-type mice. The results provide further information about the important roles of the sequences encoded within exon 1 and exons 2 and 3 of mu-opioid receptor gene for the activation of G-proteins by mu-opioid receptor agonists in the mouse spinal cord. PMID- 14737013 TI - Expression of human organic anion transporters in the choroid plexus and their interactions with neurotransmitter metabolites. AB - The purpose of the present study was to elucidate the expression of human organic anion transporter 1 (hOAT1) and hOAT3 in the choroid plexus of the human brain and their interactions with neurotransmitter metabolites using stable cell lines. Immunohistochemical analysis revealed that hOAT1 and hOAT3 are expressed in the cytoplasmic membrane and cytoplasm of human choroid plexus. Neurotransmitter metabolites, namely, 5-methoxyindole-3-acetic acid (5-MI-3-AA), homovanillic acid (HVA), vanilmandelic acid (VMA), 3,4-dihydroxyphenylacetic acid (DOPAC), 5 hydroxyindole-3-acetic acid (5-HI-3-AA), N-acetyl-5-hydroxytryptamine (NA-5-HTT), melatonin, 5-methoxytryptamine (5-MTT), 3,4-dihidroxymandelic acid (DHMA), 5 hydroxytryptophol, and 5-methoxytryptophol (5-MTP), but not methanephrine (MN), normethanephrine (NMN), and 3-methyltyramine (3-MT), at 2 mM, inhibited para aminohippuric acid uptake mediated by hOAT1. On the other hand, melatonin, 5-MI-3 AA, NA-5-HTT, 5-MTT, 5-MTP, HVA, 5-HI-3-AA, VMA, DOPAC, 5-hydroxytryptophol, and MN, but not 3-MT, DHMA, and NMN, at 2 mM, inhibited estrone sulfate uptake mediated by hOAT3. Differences in the IC(50) values between hOAT1 and hOAT3 were observed for DHMA, DOPAC, HVA, 5-HI-3-AA, melatonin, 5-MI-3-AA, 5-MTP, 5-MTT, and VMA. HOAT1 and hOAT3 mediated the transport of VMA but not HVA and melatonin. These results suggest that hOAT1 and hOAT3 are involved in the efflux of various neurotransmitter metabolites from the cerebrospinal fluid to the blood across the choroid plexus. PMID- 14737014 TI - Acquired nasal hyperresponsiveness aggravates antigen-induced rhinitis in the guinea pig. AB - Whether a state of nasal hyperresponsiveness influences antigen-induced biphasic nasal blockage and sneezing were examined using a guinea pig model of allergic rhinitis. Sensitized animals were challenged with an antigen, Japanese cedar pollen, once every week. Before the 13th challenge, the animals were randomly divided into 2 groups, and then the 13th challenge was performed (Groups A-0 and B-0). The 14th challenge was done on day 2 (Group A-2) and on day 7 (Group B-7) after the 13th challenge, on which nasal hyperresponsiveness was present and absent, respectively. Biphasic nasal blockage and sneezing after the challenge in Group A-2 were more severe than those in Group A-0, while those of Group B-7 were almost the same as those of Group B-0. An anti-histaminic, mepyramine, inhibited sneezing but not the biphasic nasal blockage in Group B-7. A cysteinyl leukotriene (CysLT) antagonist, pranlukast, suppressed the late nasal blockage but not the early blockage and sneezing in Group B-7. In contrast, in Group A-2, mepyramine significantly attenuated not only sneezing but also the early nasal blockage. Pranlukast significantly inhibited both nasal blockage and sneezing in Group A-2. In conclusion, nasal hyperresponsiveness aggravated the antigen induced nasal responses, to which histamine and CysLTs considerably contributed. PMID- 14737015 TI - Effects of the serotonergic anxiolytic buspirone on plasma glucose and glucose induced hyperglycemia in mice. AB - Effects of the serotonergic anxiolytic buspirone on plasma glucose and glucose induced hyperglycemia were studied in mice. Buspirone did not affect plasma glucose levels of non-fasted mice, while it increased serum insulin levels. In fasted mice, buspirone significantly reduced glucose-induced hyperglycemia and enhanced insulin release elicited by glucose. This suggests that buspirone enhances insulin release, resulting in inhibition of glucose-induced hyperglycemia. The major metabolite of buspirone, 1-(2-pyrimidinyl)piperazine (1 PP) increased serum insulin levels and induced a slight hypoglycemia in non fasted mice. 1-PP decreases glucose-induced hyperglycemia and amplifies insulin release elicited by glucose in fasted mice. Since buspirone is mainly metabolized to 1-PP and formation of 1-PP occurs quickly, the inhibitory effect of buspirone on glucose-induced hyperglycemia is likely mediated by 1-PP. PMID- 14737016 TI - Effects of acute repetitive transcranial magnetic stimulation on extracellular serotonin concentration in the rat prefrontal cortex. AB - Repetitive transcranial magnetic stimulation (rTMS) changes the function of the cortex. This study clarified the effects of acute rTMS treatment on extracellular serotonin (5-HT) concentrations in the rat prefrontal cortex (PFC) by using in vivo microdialysis methods. Each rat received acute rTMS treatment of the frontal brain at 500 stimuli from twenty trains applied at 25 Hz for 1 s at 1-min intervals between trains. Sham-treated rats received the same handling procedure and sound of the stimulator. Sham treatment increased the extracellular 5-HT levels compared with the non-treated group. However, rTMS treatment using the stimulation intensity of 110% motor threshold eliminated the increase in 5-HT levels induced by the sham treatment. Acute rTMS treatment of the frontal brain is related to the serotonergic neuronal system in the rat PFC, and it may have therapeutic implications for emotional disorders. PMID- 14737017 TI - Anti-stress effects of Ginkgo biloba and Panax ginseng: a comparative study. AB - Stress is a global menace fortified by the advancement of industrialization. Failure of stress management is due to lack of proper evaluation of anti-stress products. We explored the anti-stress potential of the Ginkgo biloba (G. biloba, 30 mg/kg, p.o.) and compared it with that of Panax ginseng (P. ginseng, 100 mg/kg, p.o.) against acute stress (AS) and chronic stress (CS) models in rats. Immediately after AS and CS, the rats were sacrificed, and adrenal glands and stomach were dissected out for weight determination and scoring of the ulcer index (UI), respectively, as well as changes in biochemical parameters like plasma glucose (GL), triglycerides (TG), cholesterol (CL), creatine kinase (CK), and serum corticosterone (CORT) were also estimated. AS significantly increased UI, adrenal gland weight (AGW), GL, CK activity, and CORT, whereas G. biloba significantly reduced them. P. ginseng significantly reverted GL and CK activity. In CS, a significant increase was found in the UI, AGW, CK activity, and CORT with a decrease in the level of CL and TG. G. biloba did not produce any significant effect on CS-induced alterations. P. ginseng reduced the UI, AGW, plasma GL, TG, CK activity, and CORT level significantly. From the above study, G. biloba is more effective in AS, whereas for CS, P. ginseng will be a better option. Hence these extracts possess significant anti-stress properties and can be used for the treatment of stress-induced disorders. PMID- 14737018 TI - Cough reflex induced by microinjection of citric acid into the larynx of guinea pigs: new coughing model. AB - We developed a new coughing model that evoked coughs by microinjection of citric acid into the larynx in unanesthetized unrestrained guinea pigs; additionally, we recorded synchronous sounds and waveforms of coughing utilizing built-in microphones and a whole body plethysmograph. The coughing model was able to distinguish a coughing response from other expiratory responses, such as an expiratory reflex or a sigh, by examining the waveform of the expiratory response and the existence of sound. It was not necessary to distinguish a cough from a sneeze, since the administration site was restricted to the larynx. Microinjection of 0.4 M citric acid, total of 20 microl (10 times, 2 microl at 30 s intervals), induced coughs (27.03 +/- 4.03 coughs in 10-min observation) that were stable and independent of the inhalation volume. In the inhalation studies, animals were exposed to citric acid only once, because the number of coughs remarkably decreased with repeated administration at intervals of 24 h (tachyphylaxis). However our coughing model was able to repeatedly challenge the microinjection of citric acid at an interval of 24 h. These results indicated that this coughing model was highly sensitive and correctly assessed the cough response. PMID- 14737019 TI - Possible role of interleukin-6 in PC12 cell death induced by MPP+ and tetrahydroisoquinoline. AB - Interleukin (IL)-6 has been shown to protect neuronal cells from cell death induced by various stimulants. Although neuronal cells including PC12 cells were shown to produce IL-6, little is known about the effects of dopaminergic neurotoxins, 1,2,3,4-tetrahydroisoquinoline (TIQ) and 1-methyl-4-phenylpyridinium ion (MPP(+)), on IL-6 expression in PC12 cells. In the present study, we investigated the role of IL-6 in the TIQ- and MPP(+)-induced cell death in PC12 cells. Treatment with 3.2 mM TIQ for 24 h caused a delayed cell death (lactate dehydrogenase (LDH) leakage and nuclear DNA fragmentation) markedly 72 h after the addition. Addition of 0.4 mM MPP(+) caused LDH leakage and nuclear DNA fragmentation 24 h after the addition. The cell death induced by MPP(+) was inhibited by an inhibitor of caspases, z-Val-Ala-Asp(OMe)-fluoromethylketone. The cell death induced by TIQ or MPP(+) was inhibited by nerve growth factor and 10% serum and significantly enhanced by the treatment with anti-IL-6 antibody. Both neurotoxins decreased the IL-6 mRNA level in PC12 cells without changing the other tested mRNA levels (IL-1 alpha, beta-actin, etc.). These findings suggest that dopaminergic neurotoxins cause cell death in PC12 cells at least partially by changing IL-6 expression. PMID- 14737020 TI - Altered mRNA expression of ATP-sensitive and inward rectifier potassium channel subunits in streptozotocin-induced diabetic rat heart and aorta. AB - Cardiovascular diseases are the most frequent and costly complication of diabetes. Many previous studies showed that ATP-sensitive potassium channels (K(ATP)) and inward rectifier potassium channels (Kir) play important regulatory roles in functions of cardiovascular tissues. It's still not very clear how these potassium channels are involved in cardiovascular complications of diabetes. We used the streptozotocin (STZ)-induced diabetic rats model to study the expressions of K(ATP) and Kir channel subtypes in diabetic cardiovascular tissues. The mRNA expression levels of Kir2.1, Kir3.1, Kir6.1, Kir6.2, and sulfonylurea receptor (SUR) 2A and 2B subunits in heart and aortal smooth muscles were determined by the reverse-transcription polymerase chain reaction. The results showed that in comparison with the control rats, mRNA expression of SUR 2A was reduced significantly in the diabetic heart (SUR 2A/GAPDH, 1.04 +/- 0.16 vs 0.38 +/- 0.09, P<0.01, n = 3); SUR 2B was reduced markedly in the aortal smooth muscle of diabetic rats (SUR 2B/GAPDH, 1.13 +/- 0.14 vs 0.35 +/- 0.07, P<0.01, n = 3). However, there are no significant expression changes of Kir2.1, Kir3.1, Kir6.1, and Kir6.2 in diabetic rats. These results suggested that expression of specific K(ATP) channel subunits were altered in the heart and aorta of diabetic rats. PMID- 14737021 TI - Effects of OPC-51803, a novel, nonpeptide vasopressin V2-receptor agonist, on micturition frequency in Brattleboro and aged rats. AB - We assessed the effects of OPC-51803 ((5R)-2-[1-(2-chloro-4-(1 pyrrolidinyl)benzoyl)-2,3,4,5-tetrahydro-1H-1-benzazepin-5-yl]-N isopropylacetamide), a nonpeptide vasopressin V(2)-receptor agonist, on micturition frequency in female homozygous Brattleboro rats (strain carries hereditary diabetes insipidus) and aged male Sprague-Dawley rats with polyuria. Female homozygous Brattleboro rats exhibited more diuresis and a larger micturition frequency over a 24-h period than did the heterozygous controls. In Brattleboro rats, an oral administration of OPC-51803 at 0.03 and 0.3 mg/kg significantly decreased urinary frequency and was accompanied by decreased urine volume. However, little effect was seen in the mean and maximal micturition volume. Aged male Sprague-Dawley rats (25-month-old) showed a significant increase in urine volume throughout a 0- to 24-h period compared with mature (6 month-old) rats. Orally administered OPC-51803 at 0.3 mg/kg decreased not only urine volume but also urinary frequency in aged rats. Furthermore, OPC-51803 prolonged the time prior to the first micturition. Therefore, OPC-51803 decreased micturition frequency in both rat species by reducing urine outflow. This suggests that the compound will be useful for treating micturition disorders that result in frequent micturition, such as that from polyuria, nocturnal polyuria, and some kinds of urinary incontinence. PMID- 14737022 TI - N-nitrosocarbofuran induces apoptosis in mouse brain microvascular endothelial cells (bEnd.3). AB - In this study, we investigated whether carbofuran, a commonly used carbamate pesticide, and N-nitrosocarbofuran (NOCF), the N-nitroso metabolite of carbofuran, have cytotoxicity in mouse brain microvascular endothelial cells (bEnd.3). Results from the MTT assay in bEnd.3 cells showed that NOCF but not carbofuran caused a remarkable decrease in cell viability. The cell death induced by NOCF appeared to involve apoptosis, based on our results from annexin V staining and electron microscopy. To investigate the mechanism of the NOCF induced cell death, we examined the effects of selective inhibitors for MAP kinase pathways, PD98059 (for MEK/ERK), SB202190 (for p38 MAP kinase), and SP600125 (for JNK), on the NOCF-induced cell death. The NOCF-induced cell death was significantly reduced by PD98059, but not by SB202190 or SP600125. NOCF increased ERK phosphorylation as early as 15 min after the treatment and this increase was maintained for 2 h. In summary, our results suggest that NOCF can induce apoptotic cell death, at least in part, through the ERK pathway in brain microvascular endothelial cells. PMID- 14737023 TI - Cell-transforming activity of fourteen chemical agents used in dental practice in Syrian hamster embryo cells. AB - Fourteen chemical agents used in dental practice were assessed for their cell transforming activity using the Syrian hamster embryo (SHE) cell transformation assay system. The cell-transforming activity was quantitatively assessed by the frequency of morphological transformation (MT) in SHE cells induced by these agents. MT was induced by m-cresol, guaiacol, formaldehyde, sodium hypochlorite, hydrogen peroxide, sodium arsenite, acid fuchsin, and basic fuchsin, but not by p chlorophenol, p-phenolsulfonic acid, glutaraldehyde, and erythrosine B. Iodine and chlorhexidine exhibited positive and pseudopositive responses, respectively. The chemical agents exhibiting a negative or pseudopositive response neither induced nor enhanced MT even in the presence of exogenous metabolic activation. PMID- 14737024 TI - Metformin protects against carbon tetrachloride hepatotoxicity in mice. AB - In the present study, the hepatoprotective effect of metformin (Met), a dimethylbiguanide anti-hyperglycemic, was examined in a mouse model of liver damage induced by chronic repeated administration of carbon tetrachloride (CCl(4)) (5 microl/kg, twice a week for 12 weeks). Met, when given orally in drinking water at an estimated daily dose of 25 or 50 mg/kg for 10 weeks starting 2 weeks after CCl(4) challenge, protected against CCl(4) hepatotoxicity. The results indicate that the hepatoprotection afforded by Met treatment at a dose of 25 mg/kg against CCl(4) toxicity may at least in part be mediated by the enhancement of mitochondrial glutathione redox status. PMID- 14737025 TI - Effects of 4,4'-diisothiocyanato-stilbene-2,2'-disulfonic acid (DIDS) and chlorpromazine on NO3- transport via anion exchanger in erythrocytes: inertness of DIDS in whole blood. AB - We examined the effects of chlorpromazine on NO(3)(-) transport between erythrocytes (RBCs) and extracellular fluid. Chlorpromazine (10 microg/ml) did not influence NO(3)(-) movement in both whole blood and RBC suspension. Though an anion exchanger (AE1) inhibitor DIDS (4,4'-diisothiocyanato-stilbene-2,2' disulfonic acid, 100 microM) did not alter NO(3)(-) movement in whole blood, it inhibited the movement in a concentration-dependent manner in the RBC suspension. The inhibition was abrogated by plasma and albumin concentration-dependently. Our results indicated that chlorpromazine had no effect on NO(3)(-) transport through AE1 and that the inertness of DIDS on AE1 in whole blood is due to interference by albumin in plasma. PMID- 14737026 TI - Strategic planning for career development. PMID- 14737027 TI - Engaging staff in nursing recruitment and retention initiatives: a multihospital perspective. PMID- 14737028 TI - Improving nurse managers' interviewing skills. PMID- 14737029 TI - Do we really practice relationship-based care? PMID- 14737030 TI - The Meaningful Retention Strategy Inventory: a targeted approach to implementing retention strategies. AB - The present and projected shortage of registered nurses mandates that administrators implement workplace incentives to retain current staff. Although several articles and studies exist on job satisfaction among nurses, few have examined retention strategies. The authors developed, tested, and implemented a tool, the Meaningful Retention Strategy Inventory, in a multihospital system. Results from the Meaningful Retention Strategy Inventory were used to guide decisions in the implementation of site-specific retention strategies. PMID- 14737031 TI - Understanding RN and unlicensed assistive personnel working relationships in designing care delivery strategies. AB - How a registered nurse and unlicensed assistive personnel partner together has implications for care delivery and, ultimately, patient outcomes. The authors summarize findings from a study that examined the characteristics of registered nurse and unlicensed assistive personnel working relationships and the care delivery practices that influence those relationships. Strategies are recommended for deploying registered nurses and unlicensed assistive personnel to promote collaboration and to improve patient care delivery. PMID- 14737032 TI - A partnership between nursing education and practice: using an empowerment model to retain new nurses. AB - A school of nursing and a health system in Massachusetts developed a unique partnership to create a system to attract and retain new nurses in acute care. The structure of this partnership was designed to increase the faculty of the school of nursing and add to the educational expertise in the department of staff development in the health system's major acute care hospital. The process was developed using an empowerment model. The authors describe the structure, process, and outcomes of this partnership. PMID- 14737033 TI - Measurement of organizational culture and climate in healthcare. AB - Although there is increasing interest in the relationship between organizational constructs and health services outcomes, information on the reliability and validity of the instruments measuring these constructs is sparse. Twelve instruments were identified that may have applicability in measuring organizational constructs in the healthcare setting. The authors describe and characterize these instruments and discuss the implications for nurse administrators. PMID- 14737034 TI - Nurse staffing models, nursing hours, and patient safety outcomes. AB - BACKGROUND DATA: Limited research has been conducted examining the effect of nurse staffing models on costs and patient outcomes. OBJECTIVE: The objective of this study was to evaluate the effect of different nurse staffing models on costs and the patient outcomes of patient falls, medication errors, wound infections, and urinary tract infections. METHODS: A descriptive correlational study was conducted in all of the 19 teaching hospitals in Ontario, Canada. The sample comprised hospitals and adult medical, surgical, and obstetric inpatients within those hospitals. RESULTS: The lower the proportion of professional nursing staff employed on a unit, the higher the number of medication errors and wound infections. The less experienced the nurse, the higher the number of wound infections. Nurse staffing models that included a lower proportion of professional nursing staff in the mix used more nursing hours in this study. CONCLUSIONS: The results of this study suggest that a higher proportion of professional nurses in the staff mix (RNs/RPNs) on medical and surgical units in Ontario teaching hospitals are associated with lower rates of medication errors and wound infections. Higher patient complexity was associated with greater patient use of nursing care resources. PMID- 14737035 TI - Community in the workplace: a proven retention strategy. AB - People yearn for a return to community, seeking a sense of connection with others. Community in the workplace is increasingly important, because many of us spend more time at work than we do anywhere else! The authors discuss community at work and the stages of formation that true communities experience. Practical helpful strategies for the nurse leader are shared. PMID- 14737036 TI - Nitric oxide modulates microglial activation. AB - BACKGROUND: Nitric oxide (NO) has important physiological regulatory roles, i.e, vasodilation, neurotransmitter release, etc. Little is known about the processes in neural tissues, which stabilize microglia. This study attempts to answer this question by demonstrating a role for basal NO in maintaining microglia juxtaposed to neurons. MATERIAL/METHODS: Mytilus edulis (a marine bivalve), were used to examine microglia egress from excised pedal ganglia microscopically. Nitric oxide is measured in excised pedal ganglia amperometrically in real-time. RESULTS: Pedal ganglia exhibit basal NO release (1 nM range). Inhibition of basal NO release by L-NAME results in greater numbers of microglia in the incubation medium. This process appears to involve two phases of egress. The first involves a slow egress of microglia, whereas the second, occurring 18 hours later, involves a more rapid release of these cells. Low levels of the NO donor SNAP (1 nM) does not interrupt microglial egress, whereas in the presence of L-NAME it does. Exposing the ganglia to high NO levels for a short period of time inhibits their egress. CONCLUSIONS: Spontaneous ganglionic NO release maintains/stabilizes microglia juxtaposed to neurons. Excised ganglia at the various observation periods reveals a transition of constitutive nitric oxide synthase (NOS) to inducible NOS derived NO. It also appears that the microglia in some unknown manner become insensitive to iNOS derived NO since they exhibit enhanced migration during this last phase of the ganglionic NO response. Taken together, NO is involved with regulating microglial activation. PMID- 14737037 TI - Catalytic nucleotide-hydrolyzing antibodies in milk and serum of clinically healthy human mothers. AB - BACKGROUND: In humans, pregnancy and lactation are associated with the production of catalytically active antibodies (abzymes) in serum and breast milk. However, the substrate specificities of the abzymes in these biological fluids, particularly breast milk, have not been studied MATERIAL/METHODS: IgG fractions were isolated from human milk by subsequent steps of chromatographic purification on Protein-A Sepharose, DEAE-cellulose, and anti-IgG Sepharose. The nucleotide hydrolyzing activity of electrophoretically homogeneous IgG antibodies was measured using 32P-labeled nucleotides and TLC. RESULTS: We demonstrated by different methods that IgG antibodies from the serum and milk of clinically healthy human mothers are able to hydrolyze ribo- and deoxyribonucleoside-5' mono, di- and triphosphates; this nucleotide-hydrolyzing activity was also present in Fab fragments of the IgG molecule. Affinity modification of the milk IgG oligomeric form by chemically reactive derivatives of ATP led to preferential modification of the L-chain. However, after separation of the subunits by SDS electrophoresis, an in-gel assay showed ATP-hydrolyzing activity in various oligomeric forms of IgG subunits (H2L2, H2L and HL), while the separated heavy (H) and light (L) chains were not catalytically active. The Km and Vmax values characterizing the interaction of IgG with nucleotides were estimated. CONCLUSIONS: Our findings speak in favor of the generation of a variety of polyclonal nucleotide-hydrolyzing antibodies by the immune system of clinically healthy mothers. PMID- 14737038 TI - Effects of gastrectomy or colectomy on liver metabolism and liver morphology in an experimental rat model. AB - BACKGROUND: It is sometimes necessary to translocate part of the gastrointestinal tract into the site of the organ earlier resected. This occurs after total esophageal, stomach or colon resection, which can lead to disturbances in the anatomical and functional balance of the gastrointestinal tract and the body as a whole. The aim of this study was to evaluate the effects of different resective surgical procedures performed on the proximal and distal GI tract in terms of total body weight, selected liver function tests, and ultrastructural changes in the liver in laboratory rats. MATERIAL/METHODS: The total of 57 resective procedures consisted of 31 total gastric and 26 total colon resections. The control group consisted of 12 animals on which only laparotomy was performed. Body weight, stool character, and the general condition of the animals were evaluated from the time of the operation. Twenty-six blood samples were collected for analysis of hemoglobin, total plasma protein, and liver aminotransferases levels. Thirty liver tissue samples from the 3 groups were taken and examined. RESULTS: We observed statistically significant decreases in total body weight, hemoglobin, and total plasma protein levels in the group where reconstructive procedures following gastrectomy were performed. In the group in which reconstructive procedures followed colectomy, a statistically significant decrease in total body weight was observed. Aminotransferase level changes were neither characteristic nor significantly different in any of the three groups involved in the investigation. CONCLUSIONS: Abnormalities of liver structure were observed at the ultrastructural level. Any correlation between these changes, general condition, and selected biochemical parameters requires further controlled clinical and experimental studies. PMID- 14737039 TI - Impact of alpha-tocopherol and vitamin C on endothelial markers in rats with streptozotocin-induced diabetes. AB - BACKGROUND: Overloads of reactive oxygen species in diabetes are associated with a number of pathological conditions, such as endothelial damage, which plays an important role in the initial stage of atherosclerosis. Plasma soluble thrombomodulin (sTM) and von Willebrand factor (vWF) have been found to be important markers of endothelial dysfunction. MATERIAL/METHODS: 33 male Wistar rats were used for this experiment. All rats received standard rat chow and water. 25 rats were made diabetic by intra peritoneal injection of streptozotocin (STZ) (65 mg/kg) and divided into three groups. Group I was allowed free access to food and water, Group II received 30 mg vitamin C daily, and Group III received a-tocopherol in a dose of 600 mg/kg body weight. RESULTS: At the end of the experimental period, there was significant elevation of sTM, vWF, fibrinogen, systolic blood pressure, and mean arterial blood pressure (MAP); however, there was a significant decrease in serum HDL cholesterol in the diabetic group in comparison to controls. Administration of antioxidants (vitamin C and a tocopherol) caused attenuation of the endothelial damage, as vitamin C administration caused a significant decrease in sTM, vWF, fibrinogen and increased HDL-cholesterol, while a-tocopherol caused a significant decrease in vWF and sTM. Both vitamin C and a-tocopherol caused a significant decrease in systolic blood pressure and mean arterial pressure. CONCLUSIONS: Our results suggest that administration of antioxidants may ameliorate the risk of thrombo embolism in diabetes associated with hypertension. PMID- 14737040 TI - The C677T methylenetetrahydrofolate reductase gene mutation and nephropathy in type 2 diabetes mellitus. AB - BACKGROUND: Diabetic nephropathy is a common cause of end-stage renal disease, responsible for about 35% of renal replacement therapy cases. The susceptibility to nephropathy is due to environmental and genetic factors and varies among individuals. Mutations of the methylenetetrahydrofolate reductase (MTHFR) gene have been shown to be associated with a predisposition to diabetic nephropathy in some populations. MATERIAL/METHODS: The study population consisted of 326 patients with type 2 diabetes and 170 healthy control subjects. Genotyping for the C677T mutation in the MTHFR gene was performed using polymerase chain reaction (PCR) and gel analysis. RESULTS: Allele and genotype frequencies did not differ significantly between the entire diabetic group and controls. There were statistically significant differences in genotype distribution between patients with nephropathy and those without. The diabetic nephropathy subgroup showed an increased frequency of the T allele as well as the TT genotype of the C677T mutation (p<0.05). The progression of renal failure to end-stage renal disease (ESRD) in dialyzed patients with diabetic nephropathy was influenced by this mutation. The mean time from diagnosis to the onset of ESRD was 3.6 years for patients with the TT genotype compared with 7.3 years for the CC genotype (p<0.01). CONCLUSIONS: Our findings suggest that the C677T mutation in the MTHFR gene predisposes type 2 diabetes patients to the development of diabetic nephropathy. The T allele of this mutation seems to be associated with a faster progression of nephropathy to end-stage renal failure. PMID- 14737041 TI - Treatment of huge uterine tumors thought to be benign in post-menopausal women. AB - BACKGROUND: When we find a huge uterine myoma-like tumor after menopause, we are hesitant whether to recommend surgery or not. MATERIAL/METHODS: In order to treat huge uterine tumors after menopause, we surgically removed a total of 25 uterine tumors over 10 cm in diameter from 25 post-menopausal women, and examined these tumors microscopically. RESULTS: Clinical assessments of the 25 tumors before surgery indicated that 24 were myomas and one was a sarcoma, based on cytology-, sonography- and MR imaging-examinations. However, the postoperative histological results revealed 22 leiomyomas, one leiomyosarcoma, one endometrial stromal sarcoma and one endometrial carcinoma at Ic stage complicated by multiple leiomyomas. Overall, 12% of the tumors were malignant (3/25 cases) and 8.3% were false negative cases (2/24 cases; diagnosed as myoma uteri) among the cases of huge uterine tumors after CONCLUSIONS: When a huge uterine tumor after menopause is diagnosed clinically as myoma uteri, surgery should be recommended since there ia a significant chance of malignancy. PMID- 14737042 TI - The effect of eight weeks of rabeprazole therapy on nitric oxide plasma level and esophageal pH and motility and motility nitric oxide plasma level in patients with erosive esophagitis. AB - BACKGROUND: Esophageal clearance, an important pathogenetic factor in gastroesophageal reflux disease, depends mainly on motility. Motility disturbances can be secondary to gastric output reflux. Nitric oxide influences esophageal motility. The aim of this study was to determine the effect of eight weeks of gastric acid secretion suppression with rabeprazole (20 mg/day) on esophageal motility. MATERIAL/METHODS: 20 patients with erosive esophagitis were studied. At study start and two weeks after the end of therapy, we recorded the results of interview, endoscopy, gastric and esophageal mucosa biopsy, 24-h esophageal pH-metry and manometry, and NO metabolites plasma concentration, determined spectrophotometrically (OXIS). RESULTS: All patients reported improvement and remained free of symptoms two weeks later. In 60% of cases, improvement of esophageal mucosa appearance was observed in endoscopic and histological examination. In follow-up we found a significantly smaller number of acid gastrooesophageal refluxes (p<0.05), reduced DeMeester score for pH range >7 (p<0.05), and greater % of time within the esophageal pH range 6-7 (p<0.05). Other esophageal pH-metry and 24-h manometry parameters did not change significantly. NO metabolites plasma concentration increased significantly (p=0.039). CONCLUSIONS: Clinical improvement after eight weeks of therapy with rabeprazole was connected with endoscopic changes only in 60% of our patients. Rabeprazole therapy did not influence esophageal motility, despite increased plasma levels of NO metabolites. Patients with erosive esophagitis need maintenance therapy, since as soon as two weeks after the end of treatment the % of monitoring time with esophageal pH<4 was similar to study start. PMID- 14737043 TI - Laparoscopic repair of "sportsman's hernia" in soccer players as treatment of chronic inguinal pain. AB - BACKGROUND: To evaluate the clinical course of soccer players affected by chronic groin pain, who were referred for laparoscopic inguinal exploration. MATERIAL/METHODS: Thirty-five professional soccer players affected by chronic groin pain and referred for laparoscopic inguinal exploration were included in this prospective, non-randomized study. Prior to surgery, patients were assessed by history, clinical examination, pelvic X-ray, bone scanning with Technetium and ultrasound of the inguinal region. The pre-peritoneal space was visualized with a laparoscope under general anesthesia. A bilateral polypropylene mesh was attached pre-peritoneally. Recorded data included demographics, history, symptoms, surgical findings and postoperative recovery. RESULTS: All patients were male. The mean age was 24.3 years. Six patients (17.1%) complained of pain affecting their normal life. A true indirect type I hernia was identified in 4 patients (11.4%). In 28 patients (80%), a wide internal ring and peritoneal dimple were found during the procedure. No clear pathology was found in three patients (8.6%). Thirty-four patients (97.1%) returned to their normal activities after surgery. CONCLUSIONS: Laparoscopic inguinal exploration and repair of "sportsman's hernias" in soccer players was associated with rapid recovery and fast return to full training and activity. PMID- 14737044 TI - The effect of hormone replacement therapy on endothelial function in postmenopausal women with hypertension. AB - BACKGROUND: Endothelial dysfunction has been implicated in the pathophysiology of cardiovascular diseases, including arterial hypertension. The present study was undertaken to assess endothelial function in postmenopausal women with arterial hypertension receiving hormone replacement therapy and antihypertensive treatment. MATERIAL/METHODS: A group of 76 women with natural menopause and essential mild to moderate arterial hypertension entered the study. Forty women received a transdermal, combined hormone replacement therapy (HRT) of 17 estradiol and norethisterone acetate, whereas 36 served as controls. At baseline and at 3 and 12 months, all patients underwent 24-hr blood pressure monitoring and an exercise test, before which, at peak exercise, and after a 15-min recovery period venous blood was drawn to measure the level of nitrite/nitrate (NOx) according to the Griess RESULTS: At 3 and 12 months after beginning HRT, the level of NOx at rest was slightly increased, with marked individual differences in response to HRT. In women not receiving HRT, NOx did not change. In the HRT group, 52.5% at 3 months and 47.5% at 12 months had significantly increased levels compared with the baseline values (17.8+/-6.7 vs. 32.8+/-4.5 vs. 28.7+/ 1.1 Kmol/l; p=0.002). The increased NOx level in responders was associated with decreased LDL cholesterol (3.62+/-1.2 vs. 3.53+/-1.3 vs. 2.6+/-0.6 mmol/l; p=0.01). At 12 months, blood pressure values did not differ from those at baseline in either group. CONCLUSIONS: The significant increase of NOx in half of the women receiving HRT suggests that only responders experience the cardioprotective effects of HRT. PMID- 14737045 TI - Factors affecting delays in diagnosis and treatment of pulmonary tuberculosis in a tertiary care hospital in Istanbul, Turkey. AB - BACKGROUND: To investigate delays in the diagnosis and treatment of inpatients with smear-positive pulmonary tuberculosis and to identify factors affecting these delays. MATERIAL/METHODS: 204 hospitalized patients with smear-positive pulmonary tuberculosis were identified. The clinical files of the patients were analyzed and questionnaires were created. RESULTS: Mean application interval was 31.4 days, mean referral interval was 22.1 days, mean diagnosis interval was 3.3 days, and mean initiation of treatment interval was 1.4 days. Patient delay was present in 34.8 percent of the patients. The application interval was shorter for patients having an index case for tuberculosis (p=0.039) and for those with good economic status (p<0.005). 167 patients (81.9%) had institutional delay. The referral interval was longer for female patients than for male patients (p=0.015). The most common causes of institutional delays were a low index of suspicion for tuberculosis, health care system delays, and underutilized chest X ray examinations. One hundred and three patients (50.5%) had delays in diagnosis and 51 patients (25.0%) had delays in treatment. The most frequent reason for diagnostic delay was health care system delays (35.9%). CONCLUSIONS: There were several delays in the diagnosis of tuberculosis patients. For an effective tuberculosis control, efforts should be made to reduce these delays. Physicians and the public should be educated about tuberculosis. Health care system and laboratory delays should be improved. PMID- 14737046 TI - Parameters of the functional and morphological status of the upper digestive tract in alcohol-dependent male patients with depression and alexithymia in the context of autonomic nervous system activity and nitric oxide plasma level. AB - BACKGROUND: Psychosomatic factors play an important role in the pathogenesis of many diseases. The aim of this study was to compare the results of upper digestive tract examinations in alcoholics with depression and/or alexithymia diagnosed with atypical chest pain. MATERIAL/METHODS: In 52 alcohol dependent male patients with atypical chest pain we performed gastroduodenoscopy, esophageal and gastric pH-metry, 24-h esophageal manometry, treadmill stress test, Holter monitoring, and blood sampling. Depression was diagnosed using the Beck Depression Inventory (BDI), and alexithymia according to the Toronto Alexithymia Scale (TAS). RESULTS: Depression and/or alexithymia were diagnosed in 37 patients (71%). Depressive and alexithymic patients, in comparison to subjects with lower BDI and TAS scores, had less intensive Helicobacter pylori antral colonization, higher values of gastro-esophageal acid reflux parameters in 24h pH metry, higher mean amplitude of contraction, longer mean contraction duration, higher percentage of hypertensive contractions, and a greater percentage of effective peristalsis, in spite of a similar percentage of peristaltic and simultaneous contractions. The patient groups did not differ in relation to gastric pH-metry parameters or factors potentially connecting mental status and esophageal function regulation, such as plasma nitric oxide metabolite concentration and heart rate variability (HRV), which correlate with autonomic nervous system activity. CONCLUSIONS: In alcoholic patients with depression and alexithymia there is a tendency to greater functional disturbances in the upper digestive tract. The observed differences between groups were not related to changes in ANS activity and nitric oxide metabolism. PMID- 14737047 TI - Violence in an urban community from the perspective of an accident and emergency department: a two-year prospective study. AB - BACKGROUND: Information about violence in a given community is usually based on crime statistics. The aim of this study was to explore violence in an urban community from the perspective of an accident and emergency department. MATERIAL/METHODS: All assault victims treated at the Bergen Accident and Emergency Department (AED) during a two-year period (1994-1996) were prospectively registered, and data were collected about the patients and the assault incidents. To assess the proportion of unrecognized assault victims treated at the AED, an anonymous questionnaire was sent to all adult patients (first-time consultations) who visited the AED during a ten-day period in 1997. RESULTS: 1803 assault victims were registered, 433 of whom (24%) were females. Most of the victims were young men assaulted at public locations, under the influence of alcohol, often by unknown attackers, and frequently feeling that the attack was unprovoked (and thus defined as street violence). Few victims of child abuse or elder abuse were identified. About 40% of the females were victims of domestic violence. Non-Norwegians, unemployed, and people living in economically deprived areas of the community were over represented. A minority of the assault victims wanted to press legal charges. From the postal survey (n=1264, response rate 43%) few unrecognized victims of violence could be identified among our patients. CONCLUSIONS: An accident and emergency department registration of violence victims will mostly identify male victims of street violence. PMID- 14737048 TI - Three kinds of cosmophysical activity: links to temporal distribution of deaths and occurrence of acute myocardial infarction. AB - BACKGROUND: The purpose of this study was to check for possible links between mortality statistics in a given month, generally from all causes and specifically from selected causes, and three kinds of cosmophysical activity: solar (SA), geomagnetic (GMA), and cosmic ray (CR). We also compared the results obtained for correlation by gender. MATERIAL/METHODS: 504,243 deaths were analyzed from the National Database of Lithuania (1990-2001); 102,604 OP deaths (1988-2000) were also included in this study and compared with data obtained from neutron monitoring data and other physical sources for the same time period. RESULTS: The total monthly number of deaths for 144 consecutive months was significantly and inversely correlated with SA and positively with CR activity, but not with GMA. The number of deaths from cancer, suicide, motor vehicle accidents, and acute myocardial infarction correlated with all three physical factors. SA and GMA show inverse effects as compared to CR. Ischemci heart disease deaths correlated only with GMA, cardiovascular accidents with SA and CR. Non-cardiovascular and suicide deaths were most strongly correlated to CR activity. CONCLUSIONS: The number of deaths is significantly related to physical activity in the space environment. These relationships show differences between kinds of deaths and significant correlation for the number of acute myocardial infarctions. Gender differences in links with physical activity in space were found in some of the compared groups. PMID- 14737049 TI - A trial of intranasal ACTH(1-24) administration to a patient with isolated ACTH deficiency. AB - BACKGROUND: Isolated adrenocorticotropic hormone (ACTH) deficiency is a rare cause of secondary adrenocortical insufficiency. Normally it could be used therapeutically as an alternative to glucocorticoid treatment in these patients. We investigated the possibility of therapeutic approach as intranasal ACTH replacement therapy in patients with ACTH deficiency. CASE REPORT: A 32-year-old woman with general fatigue, weakness of legs and loss of consciousness due to severe hyponatremia was admitted to our hospital. Endocrinological studies showed low levels of plasma ACTH and serum cortisol with the loss of circadian rhythm. Plasma ACTH and serum cortisol levels failed to respond after intravenous injection of human corticotropin releasing hormone (hCRH), however, serum cortisol showed a blunted response to ACTH(1-24) stimulation test. She was diagnosed isolated ACTH deficiency. We performed continuous intranasal administration of ACTH(1-24) to the patient. There were no cortisol, aldosterone and dehydroepiandrosterone (DHEA) responses to a single intranasal ACTH(1-24) administration while these levels increased 6 days after intranasal treatment of ACTH(1-24). CONCLUSIONS: These data demonstrate that adrenocortical steroids production are stimulated by intranasal administration of ACTH(1-24) in this patient with isolated ACTH deficiency. We suggest that intranasal administration of ACTH offers a therapeutic approach as ACTH replacement therapy in patients with ACTH deficiency. The latter may be more physiologic than glucocorticoid replacement. PMID- 14737050 TI - Model of the epidemic of childhood atopy. AB - The reasons for the steep rise in the prevalence of atopic diseases such as asthma, atopic eczema and allergic rhinitis are unexplained. Studies have shown that immune regulation in the intestinal mucosa-associated lymphoid tissue (MALT) plays an important role in atopic sensitization and is influenced by the intestinal microflora. The hypothesis presented here is based on the following assumptions: 1. Mothers transfer some of their intestinal microflora to their children at birth and by close contact. 2. The intestinal microflora has the properties of an ecosystem that may react unexpectedly to changing conditions. 3. Modern lifestyle and diets negatively influence the intestinal ecosystem during the lifespan. Thus, from one generation to the next, there may be cumulative degradation of the intestinal microflora. Children inherit a non-physiological gut flora that is further degraded in later life and contributes to atopic sensitization in the following generation. Linear changes in environmental conditions and lifestyle may lead to non-linear changes in the gut flora and, possibly, to an increasing susceptibility to atopic diseases. Clinical studies using probiotics and dietary intervention should be the focus of future research. PMID- 14737051 TI - Theoretical modeling study of the necrotic field during high-intensity focused ultrasound surgery. AB - BACKGROUND: High-intensity focused ultrasound (HIFU) is a type of micro-invasive treatment of tumor. It is important to create a guideline for the volume and position of the necrotic field by means of a theoretical model, this being one of the key problems in the clinic application of HIFU. MATERIAL/METHODS: Based on cellular thermo-necrotic theory, a computational model of temperature distribution and, therefore, the necrotic field was developed. A multi-transducer system with non-interferential and self-focused property was devised to provide experimental verification of the theoretical model. RESULTS: The necrotic field resulting from the modeling simulation did not show any evident difference from that of the experiment. The necrotic field of the same heating duration (4 seconds) was found with both modeling and experiment to be spheroid, with its center at the system's geometric focus and a volume of 1x1x2 cm3. CONCLUSIONS: The model of this study may predict the practical volume of the necrotic field as well as the time needed for it to form. PMID- 14737052 TI - Correlation of transiently evoked otoacoustic emission measures to auditory thresholds. AB - BACKGROUND: The presence of otoacoustic emissions is objective evidence of normal cochlear status. However, this test cannot be used to predict absolute auditory thresholds with accuracy. The criteria of normal otoacoustic emissions are not yet standardized and different measures of otoacoustic emissions have been used in various settings. In this study, the measures of reproducibility and otoacoustic emission amplitude are compared with regard to their correlation to pure-tone auditory (PTA) thresholds. MATERIAL/METHODS: One hundred and seventeen subjects with normal hearing were included in the study. Subjects with previous audiological, otolaryngologic, or vestibular disease were excluded. A complete ENT and audiological work-up was performed, including transiently evoked otoacoustic emissions (TEOAEs). The equipment used for TEOAE testing was a DP Echoport ILO 292 Otodynamics analyzer connected to a portable personal computer. A full 260 low-noise samples were averaged. Correlation between PTA thresholds and either overall TEOAE amplitude or whole reproducibility was estimated using multiple regression analysis. RESULTS: Correlation between acoustic thresholds at 250, 500, 1000, 2000, 4000 and 8000 Hz and either whole reproducibility or emission amplitude was highly significant for both measures. However, whole reproducibility was better correlated to acoustic thresholds at 250, 500, 1000, 2000, 4000 and 8000 Hz than was emission amplitude, and was more accurately predicted from multiple regression equations. CONCLUSIONS: Reproducibility measures performed better than TEOAE amplitude levels in the prediction of auditory thresholds. PMID- 14737053 TI - Stroke volume variability and heart rate power spectrum in relation to posture changes in healthy subjects. AB - BACKGROUND: Power spectrum analysis of heart rate variability (HRV) can be used to assess the state of sympathovagal balance-modulating sinus node activity. Stroke volume variability (SVV) probably reflects the dynamics of physiological control mechanisms over time. MATERIAL/METHODS: Power spectrum HRV and SVV were simultaneously measured in 44 healthy men. The values of the HRV and SVV parameters were compared for the supine and upright body positions in three age groups (I: 20-40; II: 40-60; III: >60 years old). The total power (TP) of the very low (VLF:<0.05 Hz), low (LF: 0.05-0.15 Hz) and high (HF: 0.15-0.5 Hz) HRV and SVV frequency bands were measured. The LF/HF and lnLF/lnHF ratios for HRV and SVV were calculated. RESULTS: The upright position caused a significant decrease in HRV in the HF and lnHF bands and an increase in the LF/HF and lnLF/HF ratios in all groups. SVV analysis for the supine position revealed significant differences in lnTP and HF values (between groups I and II) and in lnHF values (between groups I and II and I and III). SVV analysis for the upright position revealed differences in lnTP (between groups I and II and I and III), lnVLF (between groups I and III and I and III), lnLF (between groups I and II and I and III) and lnHF (between groups I and II and I and III). CONCLUSIONS: The combined analysis of HRV and SVV revealed different cardiovascular responses to postural stress in the three age groups considered. PMID- 14737054 TI - Autonomic signs and dosing during the initial stages of clozapine therapy. AB - BACKGROUND: Recent reports implicate clozapine in heart rate variability, QTc prolongation, torsade de pointes and sudden death at therapeutic doses, even in physically healthy patients. This study aims to examine whether autonomic (vital) signs are correlated with clozapine dose titration and blood levels of clozapine and nor-clozapine during clozapine therapy. MATERIAL/METHODS: Thirty-seven consecutive patients with diagnosis of schizophrenia treated with clozapine were included in this prospective longitudinal study. The study was restricted to only the first 8 weeks of treatment. After obtaining informed consent, serum concentrations of clozapine and nor-clozapine were determined weekly at trough, as doses were administered q12h and adjusted according to clinical guidelines for clozapine use. Autonomic signs including BP (supine and erect), pulse (supine and erect) and temperature were monitored daily each morning before and one hour after the morning's dose of clozapine was administered. RESULTS: We calculated analyses of covariance (ANCOVAs) to evaluate the changes in vital signs parameters, from baseline to week 8, with clozapine variables as covariates (i.e, the dose of clozapine, as well as the levels of serum clozapine and nor clozapine). The blood pressure and pulse did not change significantly (p<0.01) from baseline to weeks 8. The temperature was inversely related to clozapine dose (p<0.003). Higher nor-clozapine to clozapine ratios were associated with higher BP measures (p=0.002). The magnitude of these relationships is weak (r<0.30). CONCLUSIONS: There is a tendency to autonomic dysregulation during clozapine use. This has cardiac function implications, justifying cautious dose adjustment with frequent monitoring of vital signs. PMID- 14737055 TI - G-CSF in solid tumor chemotherapy: a tailored regimen reduces febrile neutropenia, treatment delays and direct costs. AB - BACKGROUND: Current guidelines do not recommend G-CSF for patients with risk factors for neutropenia. MATERIAL/METHODS: One-hundred patients undergoing chemotherapy were randomized to treatment with G-CSF at 5 Kg/kg for established febrile neutropenia (ANC <1000/microl) (Group A) or G-CSF at 263 Kg/day if ANC was 1500/microl or less on the day of the expected nadir, with the duration of treatment determined by the severity of neutropenia (Group B). RESULTS: The number of doses of G-CSF was similar in the two groups. There were 34 cases of febrile neutropenia in Group A, but none in Group B (p=0.0001). Hospital admission for febrile neutropenia, antibiotic use and delays in chemotherapy were all significantly more common in Group A. Total direct costs were estimated to be 66, 646 for Group A and 47, 119 for Group B. CONCLUSIONS: Tailoring treatment does not increase G-CSF use, but significantly reduces febrile neutropenia and treatment delays and lowers direct costs. PMID- 14737056 TI - Pregnancy complications and perinatal outcome in diabetic women treated with Humalog (insulin lispro) or regular human insulin during pregnancy. AB - BACKGROUND: Pregnancy outcome in diabetic women is strictly related to glycemic control during pregnancy. The aim of our study was to compare pregnancy outcome between patients subjected to intensive insulin therapy using regular human insulin and those treated with insulin lispro (Humalog). MATERIAL/METHODS: Group A (n=25) was treated with Humalog, and the control group B (n=46) with regular human insulin. Mean age, duration of diabetes, presence of chronic diabetic complications (according to the White classification) parity, and BMI did not differ between groups. RESULTS: The mean HbA1c concentrations in groups A and B were respectively: 7.8+/-1.4% vs. 7.5+/-1.5% in the first trimester, 6.4+/-0.8% vs. 6.5+/-1.6% in the second, and 6.7+/-0.7% vs. 6.3+/-1.2% in the third (no significant differences). The duration of pregnancy was 36.4+/-3.9 weeks in group A and 37.1+/-1.9 weeks in group B, while the mean neonatal birth weight was 3467+/-790 and 3367+/-666 g, respectively. Neither the frequency of preterm labor and cesarean section nor the frequency of fetal macrosomia and hypoglycemia differed between groups. There was only one malformed infant in the human insulin treated group, and no statistical difference in the rate of spontaneous abortion between groups. Also, there were no differences in the frequencies of occurrence of hypertension (essential and pregnancy induced) and urinary tract infections. CONCLUSIONS: The course of pregnancy and perinatal outcome is comparable in intensively treated diabetic women regardless of the short-acting insulin used. Humalog appears to be a safe alternative to human insulin in the treatment of diabetes during pregnancy. PMID- 14737057 TI - The perils of paying academic physicians according to the clinical revenue they generate. AB - Proponents of paying academic physicians according to the clinical revenue they generate cite the dangers of failing to reward high-performing faculty members, the unfairness of equally compensating physicians who generate very different amounts of revenue, and the positive role variable compensation can play in promoting the financial health of the organization. Yet there are caveats in revenue-based compensation (RBC). Amount of clinical effort and the revenue it generates are not necessarily tightly linked. Such approaches emphasize revenue producing activities such as patient care at the expense of other academic missions such as education. By focusing attention on pay, RBC may distract our best faculty members from the intrinsic rewards of academic medicine. And by fostering financial competition within the academic health center, RBC jeopardizes mutual dedication to the larger missions of the organization as a whole. Debates over how to compensate medical school faculty members represent nothing less than a battle for the heart and soul of academic medicine. PMID- 14737058 TI - Screening for lung cancer--a review. AB - Despite recent advances in oncology lung cancer remains the most common cause of cancer death in the United States, and its cure rate has not improved for the past 20 years. Lung cancer has been a target for numerous screening strategies, aimed at its earlier detection, and potentially improved cure. We describe the research grounds for screening for malignancies, including types of bias inherent to screening trials and present a brief discussion of potential outcomes of screening. We then discuss the results of trials of chest radiography and sputum analysis. We then comment on the recent and on-going research of computed tomography (CT) screening for lung cancer. Computed tomography offers many advantages over routine radiographs in screening for lung cancer. Recent data indicate an impressive stage shift and improved resectability of lung cancers detected by the CT. Large-scale studies with longer periods of follow-up will show whether these promising results will translate into an improved lung cancer related mortality in the screened population. PMID- 14737059 TI - Strength training among adults aged >/=65 years--United States, 2001. AB - Strength training (also referred to as resistance training) enables adults to improve their overall health and fitness by increasing muscular strength, endurance, and bone density and by improving their insulin sensitivity and glucose metabolism. For older adults (i.e., persons aged >/=65 years), strength training exercises are recommended to decrease the risk for falls and fractures and to promote independent living. The American College of Sports Medicine recommends that adults include strength training as part of a comprehensive physical activity program. A national health objective for 2010 is to increase to 30% the proportion of adults who perform, >/=2 days per week, physical activities that enhance and maintain muscular strength and endurance. To determine the percentage and characteristics of older adults who perform strength training consistent with this objective, CDC analyzed data from the 2001 National Health Interview Survey (NHIS). This report summarizes the results of that analysis, which indicated that approximately 12% of persons aged 65-74 years and 10% of persons aged >/=75 years met the strength-training objective. These findings underscore the need for programs that encourage older adults to incorporate strength training into their lives along with regular physical activity. PMID- 14737060 TI - Measles mortality reduction--West Africa, 1996-2002. AB - The World Health Organization (WHO) estimates that, during 2000, measles accounted for approximately 777,000 deaths worldwide, of which 452,000 (58%) occurred in Africa. In response, in 2000, WHO's African Regional Office (AFRO) adopted a plan to reduce measles mortality >50% by 2005. The plan recommended 1) increasing measles vaccination by strengthening routine health services; 2) providing a second opportunity for measles vaccination for all children, primarily through wide--age-range supplemental immunization activities (SIAs); 3) enhancing measles surveillance; and 4) improving management of measles cases. The initial wide--age-range SIA targets all children aged 9 months-14 years, regardless of history of measles disease or vaccination. Follow-up SIAs are needed 3-5 years after the initial SIA to provide a second opportunity for vaccination to children born since the previous SIA (i.e., those aged 9 months-4 years). During the 1990s, the countries of the Americas and seven countries in southern Africa used this strategy to reduce the number of measles deaths to near zero. This report describes the recent implementation of this strategy in three West African countries, where reported measles cases declined 83%-97% during the first year after SIAs. Successful implementation of this strategy by other African countries should result in achieving the goal of >50% reduction in measles mortality by 2005. PMID- 14737061 TI - Measles outbreak associated with an imported case in an infant--Alabama, 2002. AB - Local transmission of measles is rare in the United States. Since 1997, the majority of measles outbreaks have been caused by imported cases. During October 19-November 15, 2002, an outbreak of 13 confirmed cases of measles occurred, with exposure in Alabama; 11 cases were among day care attendees who had not yet been vaccinated for measles. This was the largest outbreak of measles in the United States since 1999. In response to this outbreak, the Alabama Department of Public Health (ADPH) and CDC conducted an epidemiologic investigation that determined the outbreak was initiated by an imported case in an infant aged 9 months who had returned recently from the Philippines. Health-care providers should continue to include measles in differential diagnoses for febrile rash illnesses in infants, particularly those with recent travel to areas where measles is endemic. PMID- 14737062 TI - Human death associated with bat rabies--California, 2003. AB - Rabies is a rapidly progressive, incurable viral encephalitis that is, with rare exception, transmitted by the bite of an infected mammal. On September 14, 2003, a previously healthy man aged 66 years who resided in Trinity County, California, died from rabies approximately 6 weeks after being bitten by a bat. This report summarizes the investigation by the Trinity and Shasta County Health Departments and the California Department of Health Services (CDHS). Persons should avoid direct contact with bats; however, if such contact occurs, the exposed person should visit a health-care provider immediately, and the exposure should be reported to local public health officials. PMID- 14737063 TI - Update: influenza activity--United States, January 11-17, 2004. AB - The number of states reporting widespread influenza activity continued to decrease during the reporting week of January 11-17, 2004. Health departments in five states reported widespread influenza activity. A total of 31 states and New York City reported regional activity, eight states reported local activity, and sporadic activity was reported by six states, the District of Columbia, Guam, and Puerto Rico. The percentage of outpatient visits for influenza-like illness (ILI) continued to decrease in all surveillance regions during the week ending January 17. For the first time since the reporting week ending November 8, 2003, the national percentage for ILI (2.0%) declined below the national baseline of 2.5%. The percentage of specimens testing positive for influenza also decreased, but the percentage of deaths attributed to pneumonia and influenza (P&I) was unchanged. PMID- 14737064 TI - Identification of a fourth locus (EVR4) for familial exudative vitreoretinopathy (FEVR). AB - PURPOSE: Familial exudative vitreoretinopathy (FEVR) is a genetically heterogeneous inherited blinding disorder of the retinal vascular system. To date three loci have been mapped: EVR1 on chromosome 11q, EVR2 on chromosome Xp, and EVR3 on chromosome 11p. The gene underlying EVR3 remains unidentified whilst the EVR2 gene, which encodes the Norrie disease protein (NDP), was identified over a decade ago. More recently, FZD4, the gene that encodes the Wnt receptor Frizzled 4, was identified as the mutated gene at the EVR1 locus. The purpose of this study was to screen FZD4 in a large family previously proven to be linked to the EVR1 locus. METHODS: PCR products were generated using genomic DNA from affected family members with primers designed to amplify the coding sequence of FZD4. The PCR products were screened for mutations by direct sequencing. Genotyping was performed in all available family members using fluorescently labeled microsatellite markers from chromosome 11q. RESULTS: Sequencing of the EVR1 gene, FZD4, in this family identified no mutation. To investigate this family further we performed high-resolution genotyping with markers spanning chromosome 11q. Haplotype analysis excluded FZD4 as the mutated gene in this family and identified a candidate region approximately 10 cM centromeric to EVR1. This new FEVR locus is flanked by markers D11S1368 (centromeric) and D11S937 (telomeric) and spans approximately 15 cM. CONCLUSIONS: High-resolution genotyping and haplotype analysis excluded FZD4 as the defective gene in a family previously linked to the EVR1 locus. The results indicate that the gene mutated in this family lies centromeric to the EVR1 gene, FZD4, and is also genetically distinct from the EVR3 locus. This new locus has been designated EVR4 and is the fourth FEVR locus to be described. PMID- 14737066 TI - Expression of transthyretin and retinol binding protein mRNAs and secretion of transthyretin by cultured monkey retinal pigment epithelium. AB - PURPOSE: To document the expression of mRNA for transthyretin (TTR) and retinol binding protein (RBP) in native and cultured Rhesus monkey retinal pigmented epithelium (RPE); to compare mRNA transcripts for these two proteins expressed in RPE with those found in whole monkey liver and brain; to demonstrate the secretion of TTR by RPE during short-term maintenance in a protein-free, defined medium, as a manifestation of the differentiated state of these cells in vitro. METHODS: Total RNA was isolated from cultured RPE in first passage, after incubation for eight days in defined, protein-free medium. Conditioned medium was collected for western analysis at this time. Total RNA was also extracted from RPE/choroid freshly dissected from monkey eyes. Using cDNA probes for human TTR and RBP, northern analysis was performed on the total RNA from fresh and cultured RPE samples, together with poly(A+) mRNA purified from monkey liver and brain. RESULTS: Conditioned medium from RPE yielded TTR protein of the expected monomer subunit molecular size. The TTR secreted de novo from the cultured cells was detectable in the absence of biosynthetic labeling. With the exception of some extremely low abundance transcripts expressed in cultured RPE, all samples contained a single 900 bp transcript for TTR. Based on relative amounts of actual message, RPE ranks higher than liver in abundance of TTR mRNA. In contrast, both native monkey RPE and cultured RPE cells expressed comparatively low levels of mRNA for RBP. All samples displayed a single RBP mRNA transcript at 1100 bp. CONCLUSIONS: Our results indicate that TTR is a significant gene product of the RPE, and may be considered as a marker for a differentiated phenotype for these cells in culture. There is increased recognition of various forms of ocular pathology associated with mutations or other malfunctions involving TTR and RBP, warranting a greater understanding of mechanisms of transcriptional and translational control for these two proteins. PMID- 14737065 TI - Diverse NF-kappaB expression in epiretinal membranes after human diabetic retinopathy and proliferative vitreoretinopathy. AB - PURPOSE: Formation of epiretinal membranes (ERMs) after proliferative diabetic retinopathy (PDR) and proliferative vitreoretinopathy (PVR) results in progressive deterioration of vision, but its pathogenic mechanisms are still unknown. This study was conducted to examine the role of nuclear factor kappa B (NF-kappaB) in the formation of ERMs after PDR and PVR. METHODS: ERM samples were obtained by vitrectomy from 10 patients with PDR (aged 53+/-12 years with 14+/-5 years of diabetes), 20 patients with PVR, and 17 patients with idiopathic ERMs. Ten PVR and 17 idiopathic ERM samples were processed for reverse transcription polymerase chain reaction (RT-PCR) analysis. In addition, 10 PDR and 10 PVR membranes were processed for immunohistochemical analysis. RESULTS: NF-kappaB mRNA expression levels were significantly higher (10 of 10 versus 9 of 17 subjects in idiopathic ERM, p=0.0119) in PVR subjects. Immunohistochemical analysis showed NF-kappaB protein expression in 8 of the 10 PDR samples as well as all 10 PVR samples, and NF-kappaB positive cells were partially double labeled with glial cell markers. Interestingly, NF-kappaB protein was also overlapped with angiogenic factor interleukin-8 (IL-8) in glial cells as well as vascular endothelial cells. CONCLUSIONS: These results suggest that NF-kappaB is involved in the formation of both glial and vascular endothelial cell components, and that these two cell types might have functional interactions that lead to the enlargement of intraocular proliferative membranes. PMID- 14737067 TI - Natriuretic peptide system in the human retina. AB - PURPOSE: The natriuretic peptide (NP) family includes atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP), and C-type natriuretic peptide (CNP). Natriuretic peptides are known to inhibit vascular cell growth and regulate vessel tone. There is also much evidence to suggest they modulate vascular permeability and angiogenesis, as well as regulating aqueous humor production in the eye. All these data indicate that the natriuretic peptide system might be involved in the development of diabetic retinopathy and glaucoma. Given the expression pattern of natriuretic peptides (NPs) and their receptors, natriuretic peptide receptor A (NPRA), natriuretic peptide receptor B (NPRB) and natriuretic peptide receptor C (NPRC) in the human retina has not yet been established, the present study was designed to determine ANP, BNP and CNP gene expression and localize the mature peptides in this tissue. The expression pattern of the genes encoding the different NP receptor subtypes was also examined. METHODS: Eyes (n=10) from human donors with no history of eye disease were fixed and processed for routine paraffin embedding. The cellular location of the NPs was established by immunohistochemistry. Real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR) was used to evaluate the expression of NP and NP receptor genes in neural retinas obtained from the contralateral eyes. RESULTS: Immunohistochemistry revealed the presence of NPs in the neural retina and retinal pigment epithelium. Positive NP immunostaining was observed within the astrocytes and in their processes enveloping vessels. In the anterior portion of the optic nerve, NPs were intensely labeled in neural bundles. We were able to detect NP gene expression in the human retina. The levels of NP receptor-encoding transcripts detected indicated no significant differential expression of genes coding for the different receptor subtypes. CONCLUSIONS: Our finding that NP receptor transcripts are expressed along with ANP, BNP, and CNP mRNA in the human retina provides evidence for a local system in this tissue. The expression of NPs in neural retinal, glial, and vascular elements of the normal adult retina suggests a role for these peptides in maintaining both the neural and vascular integrity of the mature retina. PMID- 14737068 TI - Differential distribution of fibroblast growth factor receptors (FGFRs) on foveal cones: FGFR-4 is an early marker of cone photoreceptors. AB - PURPOSE: Relatively little is known of the expression and distribution of FGF receptors (FGFR) in the primate retina. We investigated expression of FGFRs in developing and adult Macaca monkey retina, paying particular attention to the cone rich, macular region. METHODS: One fetal human retina was used for diagnostic PCR using primers designed for FGFR1, FGFR2, FGFR3, FGFR4, and FGFR like-protein 1 (FGFrl1) and for probe design to FGFR3, FGFR4, and FGFrl1. Rat cDNA was used to synthesize probes for FGFR1 and FGFR2 with 90% and 93% homology to human, respectively. Paraffin sections of retina from macaque fetuses sacrificed at fetal days (Fd) 64, 73, 85, 105, 115, 120, and 165, and postnatal ages 2.5 and 11 years were used to detect FGF receptors by immunohistochemistry and in situ hybridization. RESULTS: PCR showed each of the FGF receptors are expressed in fetal human retina. In situ hybridization indicated that mRNA for each receptor is expressed in all retinal cell layers during development, but most intensely in the ganglion cell layer (GCL). FGFR2 mRNA is reduced in the adult inner (INL) and outer (ONL) nuclear layers, while FGFrl1 mRNA is virtually absent from the adult ONL. FGFR4 mRNA is particularly intense in fetal and adult cone photoreceptors. Immunoreactivity to FGFR1-FGFR4 was detected in the interphotoreceptor matrix in what appeared to be RPE microvilli associated with developing photoreceptor outer segments, and generally is high in the GCL and low in the INL. Different patterns of FGFR3 and FGFR4 immunoreactivities in the outer plexiform layer (OPL) suggest localization of FGFR3 to horizontal cell processes, with FGFR4 being expressed by both horizontal and bipolar cell processes. FGFR1, FGFR3, and FGFR4 immunoreactivities are present in the inner segments and somata of adult cones. The pedicles of developing and adult cones are FGFR1 and FGFR3 immunoreactive, and the basal, synaptic region is FGFR4 immunoreactive. FGFR4 labels cones almost in their entirety from early in development and is not detected in rods. The fibers of Henle are intensely FGFR4 immunoreactive in adult cones. CONCLUSIONS: The results show high levels of FGF receptor expression in developing and adult retina. Differential distribution of FGF receptors across developing and adult photoreceptors suggests specific roles for FGF signalling in development and maintenance of photoreceptors, particularly the specialized cones of the fovea. PMID- 14737069 TI - Acute promyelocytic leukemia: where does it stem from? AB - A fundamental issue in cancer biology is the identification of the target cell in which the causative molecular lesion arises. Acute myeloid leukemia (AML) is thought to reflect the transformation of a primitive stem cell compartment. The resultant 'cancer stem cells' comprise only a minor portion of the leukemic clone but give rise through differentiation to more committed progenitors as well as differentiated blasts that constitute the bulk of the tumor. The maintenance of the leukemic clone is dependent on the self-renewal capacity of the cancer stem cell compartment, which is revealed by its ability to re-initiate leukemia in a transplant setting. The cellular basis of acute promyelocytic leukemia (APL) is however less clear. APL has traditionally been considered to be the most differentiated form of AML and to arise from a committed myeloid progenitor. Here we review apparently conflicting evidence pertaining to the cellular origins of APL and propose that this leukemia may originate in more than one cellular compartment. This view could account for many apparent inconsistencies in the literature to date. An understanding of the nature of the target cell involved in transformation of APL has important implications for biological mechanism and for clinical treatment. PMID- 14737070 TI - Identification of a gene element essential for leukemia-specific expression of transgenes. AB - Leukemia-specific promoters and enhancers for gene therapy had never been reported. Since the Wilms' tumor gene WT1 is overexpressed in almost all types of leukemia, WT1 is an ideal target of leukemia-specific therapy. To explore the possibility of gene therapy for leukemia using WT1 promoter and enhancer, their activities in several kinds of cells were analyzed by using the enhanced green fluorescent protein (EGFP) gene as a reporter. First, we identified the best combination (654P/EGFP/int3- enh/3'-enh vector) of the 654-bp WT1 promoter and the two WT1 enhancers located in intron 3 and at the 3' end of the WT1 gene for inducing EGFP expression in K562 cells, which endogenously expressed WT1. When this was transfected into WT1-expressing leukemia cells (K562, HEL), WT1 nonexpressing hematopoietic cells (Daudi, U937), and WT1-expressing nonhematopoietic cells (TYK-nu-CPr, SW480, 293 T), 19.8, 22.9, 1.47, 1.43, 4.50, 4.16, and 1.09 times EGFP expression was induced, respectively, compared to that by the promoter-less EGFP vector. These results showed that the 654P/EGFP/int3 enh/3'-enh vector specifically induced high levels of EGFP expression in WT1 expressing leukemia cells. 654P/int3- enh/3'-enh vector containing transgenes such as suicide genes might become useful tools for leukemia-specific gene therapy. PMID- 14737071 TI - DNA microarray analysis of natural killer cell-type lymphoproliferative disease of granular lymphocytes with purified CD3-CD56+ fractions. AB - Natural killer (NK) cell-type lymphoproliferative disease of granular lymphocytes (LDGL) is characterized by the outgrowth of CD3(-)CD16/56(+) NK cells, and can be further subdivided into two distinct categories: aggressive NK cell leukemia (ANKL) and chronic NK lymphocytosis (CNKL). To gain insights into the pathophysiology of NK cell-type LDGL, we here purified CD3(-)CD56(+) fractions from healthy individuals (n=9) and those with CNKL (n=9) or ANKL (n=1), and compared the expression profiles of >12 000 genes. A total of 15 'LDGL-associated genes' were identified, and a correspondence analysis on such genes could clearly indicate that LDGL samples share a 'molecular signature' distinct from that of normal NK cells. With a newly invented class prediction algorithm, 'weighted distance method', all 19 samples received a clinically matched diagnosis, and, furthermore, a detailed cross-validation trial for the prediction of normal or CNKL status could achieve a high accuracy (77.8%). By applying another statistical approach, we could extract other sets of genes, expression of which was specific to either normal or LDGL NK cells. Together with sophisticated statistical methods, gene expression profiling of a background-matched NK cell fraction thus provides us a wealth of information for the LDGL condition. PMID- 14737072 TI - A novel recombinant bispecific single-chain antibody, bscWue-1 x CD3, induces T cell-mediated cytotoxicity towards human multiple myeloma cells. AB - The development of antibody-based strategies for the treatment of multiple myeloma (MM) has been hampered so far by the fact that suitable plasma cell specific surface antigens have been missing. However, recently a novel monoclonal antibody, designated Wue-1, has been generated that specifically recognizes normal and malignant human plasma cells. Therefore, Wue-1 is an interesting and promising candidate to develop novel immunotherapeutic strategies for the treatment of MM. One variant for an antibody-based strategy is the bispecific antibody approach. Recombinant bispecific single-chain (bsc) antibodies are especially interesting candidates because they show exceptional biological properties. We have generated a novel MM-directed recombinant bsc antibody, bscWue-1 x CD3, and analyzed the biological properties of this antibody using the MM cell line NCI-H929 and primary cells from the bone marrow of patients with MM. We were able to show that bscWue-1 x CD3 induces efficient and selective T-cell mediated cell death of NCI-H929 cells and primary myeloma cells in nine out of 11 cases. The bscWue-1 x CD3 Ab is efficacious even at low E:T ratios, and with or without additional T-cell pre- or costimulation. Target cell lyses were specific for Wue-1 antigen-positive cells and could be blocked by the Wue-1 monoclonal antibody. PMID- 14737073 TI - Differentially expressed genes in adult familial myelodysplastic syndromes. AB - The precise genetic events leading to myelodysplastic syndromes (MDSs) and leukemic transformation remain poorly defined. Even less is known about adult familial MDS. We report an adult MDS family in whom enriched tissue-specific transcripts were derived by subtractive hybridization of cDNA from the mononuclear and CD34+ cells of affected and unaffected family members. These expression libraries were then hybridized to Genome Discovery arrays containing 18 404 genes and expressed sequence tags, and several clusters of differentially expressed genes were identified. A group of 21 genes was underexpressed (>5-fold) in affected vs unaffected family members, and among these were transcription factors and genes involved in myeloid differentiation, such as ZNF140 and myeloid nuclear differentiation antigen (MNDA). Another group of 36 genes was overexpressed (>5-fold), and these encoded proteins belonging to signaling pathways, such as Ras- and Fos-related genes. The top two genes downregulated in this MDS family, ZNF140 and MNDA, were similarly altered in another MDS family, and in some cases of sporadic MDS. Our data suggest that we have identified genes differentially expressed in adult familial MDS, and that alteration of some of these genes may also be important for the evolution of different stages or severity of sporadic MDS. PMID- 14737074 TI - Downmodulation of ERK protein kinase activity inhibits VEGF secretion by human myeloma cells and myeloma-induced angiogenesis. AB - The mitogen-activated protein (MAP) cascade leading to the activation of extracellular signal-regulated kinases 1/2 (ERK1/2) is critical for regulating myeloma cell growth; however, the relationship of ERK1/2 activity with vascular endothelial growth factor (VEGF) production and the effects of its downmodulation in myeloma cells are not elucidated. We found that the treatment with MAP/ERK kinase 1 (MEK1) inhibitors PD98059 or PD184352 produced a reduction of phosphorylated ERK1/2 (p-ERK1/2) levels in myeloma cells of more than 80% and prevented the increase of p-ERK1/2 induced by interleukin-6 (IL-6). MEK1 inhibitors also induced a significant inhibition of myeloma cell proliferation and blunted the stimulatory effect induced by IL-6. A significant inhibition of basal VEGF secretion by myeloma cells as well as a suppression of the stimulatory effect of IL-6 on VEGF was observed by either PD98059 or PD184352. Moreover, we also found that the PI3K kinase inhibitors, but not p38 MAPK inhibitors, reduced VEGF secretion by myeloma cells and increase the inhibitory effect of MEK1 inhibitors. In an 'in vitro' model of angiogenesis, we found that MEK1 inhibitors impair vessel formation induced by myeloma cells and restored by VEGF treatment, suggesting that the downmodulation of ERK1/2 activity reduces myeloma-induced angiogenesis by inhibiting VEGF secretion. PMID- 14737075 TI - Nucleoside transporters in chronic lymphocytic leukaemia. AB - Nucleoside derivatives have important therapeutic activity in chronic lymphocytic leukaemia (CLL). Experimental evidence indicates that in CLL cells most of these drugs induce apoptosis ex vivo, suggesting that programmed cell death is the mechanism of their therapeutic action, relying upon previous uptake and metabolic activation. Although defective apoptosis and poor metabolism often cause resistance to treatment, differential uptake and/or export of nucleosides and nucleotides may significantly modulate intracellular drug bioavailability and, consequently, responsiveness to therapy. Two gene families, SLC28 and SLC29, encode transporter proteins responsible for concentrative and equilibrative nucleoside uptake (CNT and ENT, respectively). Furthermore, selected members of the expanding ATP-binding cassette (ABC) protein family have recently been identified as putative efflux pumps for the phosphorylated forms of these nucleoside-derived drugs, ABCC11 (MRP8) being a good candidate to modulate cell sensitivity to fluoropyrimidines. Sensitivity of CLL cells to fludarabine has also been recently correlated with ENT-type transport function, suggesting that, besides the integrity of apoptotic pathways and appropriate intracellular metabolism, transport across the plasma membrane is also a relevant event during CLL treatment. As long as nucleoside transporter expression in leukaemia cells is not constitutive, the possibility of regulating nucleoside transporter function by pharmacological means may also contribute to improve therapy. PMID- 14737076 TI - Abundant expression of fibronectin is a major feature of leukemic dendritic cells differentiated from patients with acute myeloid leukemia. AB - Dendritic cells (DCs) are the most potent antigen-presenting cells responsible for the initiation of primary immune responses, playing a key role in eliciting effective antitumor immune responses. We reported previously that leukemic blasts from selected patients with acute myeloid leukemia (AML) were able to differentiate in vitro into cells with DC features. In order to identify genes differentially expressed in leukemia-derived DCs (AML-DCs), a polymerase chain reaction (PCR)-based subtraction approach was applied using cDNA from AML-DCs and monocyte-derived DCs from healthy donors as competitors. In the 548 sequences analyzed, 80% corresponded to fibronectin (FN) gene fragments. Overexpression of FN in AML-DCs was demonstrated both by semiquantitative PCR analysis and by immunostaining. In addition, we could show that FN was secreted by AML-DCs. Indeed, FN overexpression was already detectable in AML blasts of M4 and M5 subtype, and was significantly induced during DC differentiation after culture. Although the molecular events leading to overexpression of FN and the in vivo relevance of this phenomenon remain to be resolved, leukemic DCs appear to have specific patterns of differentiation, warranting stringent biological and cellular monitoring for the development and testing of leukemic DC-based immunotherapeutic strategies. PMID- 14737077 TI - Acquisition of FLT3 or N-ras mutations is frequently associated with progression of myelodysplastic syndrome to acute myeloid leukemia. AB - The role of internal tandem duplication of fms-like tyrosine kinase 3 (FLT3/ITD), mutations at tyrosine kinase domain (FLT3/TKD) and N-ras mutations in the transformation of myelodysplastic syndrome (MDS) to AML was investigated in 82 MDS patients who later progressed to AML; 70 of them had paired marrow samples at diagnosis of MDS and AML available for comparative analysis. Five of the 82 patients had FLT3/ITD at presentation. Of the 70 paired samples, seven patients acquired FLT3/ITD during AML evolution. The incidence of FLT3/ITD at diagnosis of MDS was significantly lower than that at AML transformation (3/70 vs 10/70, P<0.001). FLT3/ITD(+) patients progressed to AML more rapidly than FLT3/ITD(-) patients (2.5+/-0.5 vs 11.9+/-1.5 months, P=0.114). FLT3/ITD(+) patients had a significantly shorter survival than FLT3/ITD(-) patients (5.6+/-1.3 vs 18.0+/-1.7 months, P=0.0008). After AML transformation, FLT3/ITD was also associated with an adverse prognosis. One patient had FLT3/TKD mutation (D835Y) at both MDS and AML stages. Additional three acquired FLT3/TKD (one each with D835 H, D835F and I836S) at AML transformation. Five of the 70 matched samples had N-ras mutation at diagnosis of MDS compared to 15 at AML transformation (P<0.001), one lost and 11 gained N-ras mutations at AML progression. Coexistence of FLT3/TKD and N-ras mutations was found in two AML samples. N-ras mutations had no prognostic impact either at the MDS or AML stage. Our results show that one-third of MDS patients acquire activating mutations of FLT3 or N-ras gene during AML evolution and FLT3/ITD predicts a poor outcome in MDS. PMID- 14737078 TI - PKC alpha mediates chemoresistance in acute lymphoblastic leukemia through effects on Bcl2 phosphorylation. AB - Overexpression of protein kinase C alpha (PKC alpha) promotes Bcl2 phosphorylation and chemoresistance in human acute leukemia cells. The contribution of non-Bcl2 mechanisms in this process is currently unknown. In this report, overexpression of PKC alpha was found not to affect cell proliferation, cell cycle, or activation of mitogen-activated protein kinases. The failure of PKC alpha overexpression to activate non-Bcl2 survival pathways suggested that PKC alpha-mediated chemoresistance requires Bcl2. Supporting this notion, REH/PKC alpha transfectants were found to be as sensitive to HA14-1 (a drug that targets Bcl2 function) as parental cells. In addition, HA14-1 abrogated PKC alpha's ability to protect REH cells from etoposide. These findings suggested that Bcl2 is necessary for the protective function of PKC alpha in REH cells. Since Bcl2 phosphorylation status is negatively regulated by protein phosphatase 2A (PP2A) and PP2A regulates PKC alpha, we investigated whether PKC alpha can conversely regulate PP2A. Overexpression of PKC alpha was found to suppress mitochondrial PP2A activity by a mechanism that, at least in part, involves suppressed expression of the regulatory subunit comprising the Bcl2 phosphatase (ie the PP2A/B56 alpha subunit). The ability of PKC alpha to target both Bcl2 and the Bcl2 phosphatase represents a novel mechanism for chemoresistance. PMID- 14737079 TI - Testing the prognostic model of Marin et al in an independent chronic myelogenous leukemia study group. PMID- 14737082 TI - Immunity to adeno-associated virus serotype 2 delivered transgenes imparted by genetic predisposition to autoimmunity. AB - Adeno-associated virus (AAV) is widely considered a promising vector for therapeutic gene delivery. This promise is based on previous studies assessing AAVs safety and toxicity, ability to infect nondividing cells, elicit a limited immune response and provide long-term gene expression. However, we now find that earlier studies underappreciated the degree of AAV immunogenicity as well as the extent to which genetic background, through regulation of immune responsiveness, influences the duration of gene expression and thereby the effectiveness of AAV mediated gene therapy. We evaluated antibody responses in 12 mouse strains to AAV serotype 2 (AAV2) and AAV2-expressed transgene products including green fluorescent protein (GFP), human alpha1-antitrypsin and murine interleukin-10. As expected, all immunocompetent mice administered AAV2 developed serologic evidence of immune responsiveness to the virus. However, a previously unidentified serologic prozone effect was observed suggesting that the concentrations of anti AAV2 antibodies may have historically been subject to marked underestimation. Furthermore, strains with genetic predisposition to autoimmunity (eg, NOD, NZW, MRL-lpr) specifically imparted a functionally deleterious immune response to AAV delivered transgene products. These findings suggest that more thorough studies of anti-AAV immunity should be performed, and that genetic predisposition to autoimmunity should be considered when assessing AAV efficacy and safety in humans. PMID- 14737083 TI - Inhibition of costimulation allows for repeated systemic administration of adenoviral vector in rhesus monkeys. AB - Immunogenicity of recombinant adenoviral (Ad) vectors severely hampers the clinical development of gene therapy protocols using repeated vector administrations. Inhibition of costimulation by APCs was explored as a strategy to circumvent the immune response against Ad particles. This strategy was tested in rhesus monkeys, treated transiently with chimeric anti-human CD40 and anti human CD86 antagonist monoclonal antibodies (MAbs) at the time of systemic administration of a recombinant Ad vector. After Ad vector administration in the absence of immunosuppressive treatment, transgene expression in the serum lasted about 3-4 weeks. All control animals developed a strong neutralizing antibody (NAb) response to the Ad particles, which totally prevented efficient administration of a second vector, as shown by the lack of transgene expression. Treatment with anti-CD40 and anti-CD86 chimeric MAbs delayed or blocked the development of a humoral response against Ad and the infiltration of CD8(+) lymphocytes into the liver. This resulted in (i) increased persistence of Ad transduced cells after injection of a first vector encoding a nonimmunogenic transgene, and (ii) the possibility of readministering a second Ad vector with significant efficacy. In both respects, the combined blockade of CD40 and CD86 was more efficient than treatment with anti-CD40 alone. This study shows for the first time in non-human primates that blocking CD40 and CD86 costimulatory molecules represents a promising strategy to inhibit immune responses against an Ad vector injected systemically. PMID- 14737084 TI - Expression of human coagulation factor VIII in adipocytes transduced with the simian immunodeficiency virus agmTYO1-based vector for hemophilia A gene therapy. AB - We demonstrate that transduction of adipocytes with a simian immunodeficiency virus agm TYO1 (SIVagm)-based lentiviral vector carrying the human coagulation factor VIII gene (SIVhFVIII) resulted in expression of the human FVIII transgene in vitro and in db/db mice in vivo. Cultured human adipocytes were transduced with the SIVagm vector carrying the GFP gene in a dose-dependent manner and transduction of adipocytes with SIVhFVIII resulted in efficient expression of human coagulation factor VIII (hFVIII; 320 +/- 39.8 ng/10(6) adipocytes/24 h) in vitro. Based upon successful transduction of adipocytes by SIV vectors carrying the lacZ gene in vivo in mice, the adipose tissue of db/db mice was transduced with SIVhFVIII. There was a transient appearance of human FVIII in mouse plasma (maximum 1.8 ng/ml) on day 11 after the injection. Transcripts of human FVIII transgene and human FVIII antigen also were detected in the adipose tissue by RT PCR and immunofluorescence, respectively, on day 14. Emergence of anti-human FVIII antibodies 14 days after the injection of SIVhFVIII may explain the disappearance of human FVIII from the circulation. These results suggest that transduction of the adipocytes with vectors carrying the human FVIII gene may be potentially applicable for gene therapy of hemophilia A. PMID- 14737085 TI - A minimally invasive approach for efficient gene delivery to rodent hearts. AB - Transcoronary gene delivery represents a desirable option to achieve global myocardial transgene expression but still requires aggressive surgical preparation in rodents. We therefore developed a catheter-based approach for cardiac gene transfer in the closed chest rat. A double-lumen balloon catheter was used to create aortic occlusion for specific infusion of adenoviral vectors carrying a beta-galactosidase transgene (1 x 10(11) PFU) into the coronaries. Simultaneously, venous return was obstructed by a second balloon catheter in the right atrium. To prolong viral incubation time, we induced a transient cardiac arrest (2 and 5 min) by a combination of acetylcholine and the beta-receptor antagonist, esmolol. At 72 h after transfection, the hearts showed a homogeneous and widespread beta-galactosidase expression, and the transduction efficiency increased and up to about 43% of cardiac myocytes (histochemistry) with a 400 fold increase of beta-galactosidase activity (luminescence assay) compared to sham-operated hearts. Pharmacological treatment aimed at increasing vascular permeability (SNAP and histamine) did not bring about synergistic effects on transfection efficiency. In addition, the method using high intracoronary pressure delivery (>300 mmHg) in a single-pass manner resulted in rather sparse beta-galactosidase expression in the myocardium (3-5% of cardiac myocytes). Therefore, the percutaneous gene delivery system described here provides a simple and minimally invasive procedure that represents a novel strategy for a homogeneous and highly efficient in vivo gene transfer to rodent hearts. Our results also suggest that prolongation of viral incubation time is an effective means for achieving highly efficient myocardial gene transduction. PMID- 14737086 TI - Lentiviral vectors pseudotyped with baculovirus gp64 efficiently transduce mouse cells in vivo and show tropism restriction against hematopoietic cell types in vitro. AB - The envelope glycoprotein from vesicular stomatitis virus (VSV-G) has been used extensively to pseudotype lentiviral vectors, but has several drawbacks including cytotoxicity, potential for priming of immune responses against transgene products through efficient transduction of antigen-presenting cells (APCs) and sensitivity to inactivation by human complement. As an alternative to VSV-G, we extensively characterized lentiviral vectors pseudotyped with the gp64 envelope glycoprotein from baculovirus both in vitro and in vivo. We demonstrated for the first time that gp64-pseudotyped vectors could be delivered efficiently in vivo in mice via portal vein injection. Following delivery, the efficiency of mouse cell transduction and the transgene expression is comparable to VSV-G-pseudotyped vectors. In addition, we found that gp64-pseudotyped lentiviral vectors could efficiently transduce a variety of cell lines in vitro, although gp64 showed a more restricted tropism than VSV-G, with especially poor ability to transduce hematopoietic cell types including dendritic cells (DCs). Although we found that gp64-pseudotyped vectors are also sensitive to inactivation by human complement, gp64 nevertheless has advantages over VSV-G, because of its lack of cytotoxicity and narrower tropism. Consequently, gp64 is an attractive alternative to VSV-G because it can efficiently transduce cells in vivo and may reduce immune responses against the transgene product or viral vector by avoiding transduction of APCs such as DCs. PMID- 14737087 TI - An enhanced autogene-based dual-promoter cytoplasmic expression system yields increased gene expression. AB - The relatively low levels of transfection that can be achieved by current gene delivery systems have limited the therapeutic utility of gene transfer. This is especially true for nonviral gene-delivery systems, where the levels of gene expression achieved are usually below the levels achieved by viral gene transfer systems. One strategy for increasing gene expression is to design a cytoplasmic expression system that does not require nuclear delivery for gene expression to occur. This can be achieved through the use of an autocatalytic cytoplasmic expression system using phage RNA polymerases. Here we describe cytoplasmic expression systems that yield increased levels of gene expression following in vitro transfection. We demonstrate direct evidence for an exponential, autocatalytic increase in gene expression using autogenes, as well as levels of reporter gene expression that are 20-fold higher than standard CMV-based nuclear expression systems. The development of a high-efficiency plasmid-based expression system could significantly improve the gene expression properties of nonviral gene-delivery systems, thereby increasing their clinical utility. PMID- 14737088 TI - Prevention of angiogenesis by naked DNA IL-12 gene transfer: angioprevention by immunogene therapy. AB - IL-12 is thought to induce a cytokine cascade with antiangiogenic effects mediated by IFN-gamma and angiostatic CXCR3 chemokine ligands. Naked DNA intramuscular injection of an expression vector plasmid producing IL-12 resulted in significant, well-tolerated elevation of serum IL-12 levels. Injection of the IL-12 plasmid at least 2 days, and up to 20 days, before subcutaneous injection of matrigel with angiogenic factors resulted in strong prevention of angiogenesis in both C57/bl and nude mice. A single injection of the IL-12 plasmid contemporarily with the matrigel or 2 days after resulted in partial, statistically not significant, inhibition. Control plasmid injection did not affect either angiogenesis or angiogenesis inhibition by IL-12 protein in vivo. Angiogenesis inhibition was observed in NK cell-depleted C57/bl and nude mice as well as in IFN-gamma(-/-) and CXCR3(-/-) knockout mice, indicating that NK- and/or T-cell-initiated IFN-gamma-chemokine cascades were not involved in the angiogenesis inhibition observed in vivo. Finally, IL-12 plasmid DNA gene transfer significantly prevented the growth and vascularization of highly angiogenic KS-Imm Kaposi's sarcoma and TS/A murine mammary carcinoma tumors in nude and/or syngeneic mice. These data suggest that a preventive gene therapy approach using antiangiogenic cytokines can effectively inhibit tumor angiogenesis and KS, representing an example of angioimmunoprevention. PMID- 14737089 TI - Induction of immune tolerance to a transplantation carbohydrate antigen by gene therapy with autologous lymphocytes transduced with adenovirus containing the corresponding glycosyltransferase gene. AB - Induction of tolerance to transplantation carbohydrate antigens is of clinical significance in recipients of ABO-incompatible allografts, or of xenografts. The experimental animal model used for studying such tolerance was that of alpha1,3galactosyltransferase (alpha1,3GT) knockout (KO) mice, which lacks the alpha-gal epitope (Galalpha1-3Galbeta1-4GlcNAc-R) and which can produce the anti Gal antibody against it. In contrast, wild-type (WT) mice synthesize the alpha gal epitope and are immunotolerant to it. KO lymphocytes transduced in vitro with adenovirus containing the alpha1,3GT gene (AdalphaGT) express alpha-gal epitopes. Administration of such lymphocytes into KO mice resulted in tolerization of naive and memory anti-Gal B cells. Mice tolerized by AdalphaGT transduced lymphocytes failed to produce anti-Gal following immunizations with pig kidney membranes (PKM) expressing multiple alpha-gal epitopes. This tolerance was perpetuated by transplanted syngeneic WT mouse hearts expressing alpha-gal epitopes. Transplanted WT hearts survived in the tolerized KO mice for at least 100 days, despite repeated PKM immunizations. Control mice receiving lymphocytes transduced with adenovirus lacking the alpha1,3GT gene were not tolerized, but produced anti Gal and rejected transplanted WT hearts. This study suggests that autologous lymphocytes transduced with adenovirus containing A or B transferase genes may induce a similar tolerance to blood group antigens in humans. PMID- 14737090 TI - Discordant effects of a soluble VEGF receptor on wound healing and angiogenesis. AB - Soluble receptors to vascular endothelial growth factor (VEGF) can inhibit its angiogenic effect. Since angiogenesis is involved in wound repair, we hypothesized that adenovirus-mediated gene transfer of a soluble form of VEGF receptor 2 (Flk-1) would attenuate wound healing in mice. C57Bl/6J and genetically diabetic (db/db) mice (each n=20) received intravenous (i.v.) injections of recombinant adenoviruses (10(9) PFU) encoding the ligand-binding ectodomain of VEGF receptor 2 (Flk-1) or cDNA encoding the murine IgG2alpha Fc fragment (each n=10). At 4 days after gene transfer, two full-thickness skin wounds (0.8 cm) were created on the dorsum of each animal. Wound closure was measured over 9-14 days after which wounds were resected for histological analysis. Prior to killing, fluorescent microspheres were systemically injected for quantitation of wound vascularity. Single i.v. injections of adenoviruses encoding soluble Flk-1 significantly decreased wound angiogenesis in both wild type and diabetic mice. Fluorescence microscopy revealed a 2.0-fold (wild type) and 2.9-fold (diabetic) reduction in wound vascularity in Flk-1-treated animals (p<0.05). Impairment of angiogenesis was confirmed by CD31 immunohistochemistry. Interestingly, despite significant reductions in wound vascularity, wound closure was not grossly delayed. Our data indicates that while VEGF function is essential for optimal wound angiogenesis, it is not required for wound closure. PMID- 14737091 TI - Transient foamy virus vector production by adenovirus vectors. AB - The genome of the prototype foamy virus (PFV) has been introduced into an adenoviral/PFV hybrid vector and tested for stable in vitro gene transfer. Three different adenoviruses are used to encode: (i) the PFV structural genes gag and pol (Ad-GagPolDeltaPacI); (ii) the PFV structural gene env (Ad-Env); and (iii) the PFV vector genome (Ad-MD9) encoding the transgene (the enhanced green fluorescent protein (eGFP) gene). Following cotransduction by the three adenoviruses, the target cells become transient PFV vector-producing cells, resulting in the in situ release of recombinant PFV at a titre of up to 10(3) vector particles/ml, which can then infect surrounding cells, leading to stable integration of the expression cassette. Stable eGFP expression, observed for up to 60 days (11 passages) in cells transduced with all three adenoviral vectors, was shown by PCR to be the result of PFV integration. In contrast, cells transduced with only the adenovirus encoding the PFV vector genome showed a marked decrease in eGFP expression by passage 2 (16 days post-transduction) and did not contain integrated PFV vector. In short, this paper describes the production of a hybrid vector capable of high in vitro transduction and stable transgene expression using adenovirus and PFV vectors. PMID- 14737092 TI - Improvement of dermatitis by iontophoretically delivered antisense oligonucleotides for interleukin-10 in NC/Nga mice. AB - IL-10 is overexpressed in skin lesions of atopic dermatitis (AD) patients and believed to be an important factor in the pathogenesis of the disease. Thus the regulation of IL-10 production is a potential solution for immunotherapeutic intervention in AD. We examined the topical delivery of an antisense oligonucleotide for mouse IL-10 (AS6) and the therapeutic effect on the skin lesions of NC/Nga mice, a human AD model. Using an iontophoresis system, about 30% of the applied dose of AS6 penetrated the skin and was distributed in the epidermis and upper dermis. Topically delivered AS6 decreased the levels of mRNA and protein of IL-10 in the lesions of NC/Nga mice, with no effect on IL-4 levels. The dorsal lesions of NC/Nga mice disappeared with repeated topical application of AS6. Topically delivered AS6 showed an inhibitory effect on the production of IL-10 in the skin lesions of NC/Nga mice and had a therapeutic effect on the established dermatitis. PMID- 14737093 TI - Inhibition of human non-small cell lung tumors by a c-Met antisense/U6 expression plasmid strategy. AB - c-Met is a receptor tyrosine kinase whose activation by hepatocyte growth factor (HGF) can lead to transformation and tumorigenicity in a variety of tumors. We investigated the effects of suppressing c-Met protein expression in human non small cell lung tumors. Expression plasmids containing either sense or antisense sequences of the human c-met gene were constructed under control of the U6 snRNA promoter. A U6 control plasmid was also constructed that did not contain any c met sequence. These constructs have been examined both in vitro and in an in vivo tumor xenograft model. The c-Met protein was downregulated by 50-60% in two lung cancer cell lines that were transiently transfected with the c-Met antisense versus U6 control. Tumor cells treated with the c-Met antisense construct also show decreased phosphorylation of c-Met and MAP kinase when exposed to exogenous HGF. Lung cancer cells were grown as xenografts in mice and treated by intratumoral liposome-mediated transfer of the c-Met sense, antisense or U6 control plasmids. The treatment of lung tumors with c-Met antisense versus U6 control plasmid resulted in the downregulation of the c-Met protein expression, a 50% decrease in tumor growth over a 5-week treatment period and an increased rate of apoptosis. These results suggest that targeting the HGF/c-Met pathway may be an effective novel strategy to treat lung cancer patients. PMID- 14737094 TI - Enhancement of suicidal DNA vaccine potency by delaying suicidal DNA-induced cell death. AB - DNA-based alphaviral RNA replicon vectors, also called suicidal DNA vectors, alleviate the concerns of integration or transformation related to conventional DNA vectors since suicidal DNA vectors eventually cause apoptosis of transfected cells. However, the expression of inserted genes in these vectors is transient and the potency of suicidal DNA vaccines may be compromised because of apoptotic cell death. Therefore, to enhance the immune response to the human papillomavirus type 16 (HPV-16) E7 antigen, we generated a DNA-based Semliki Forest virus vector, pSCA1, encoding E7 fused with BCL-xL, an antiapoptotic member of the BCL 2 family. Our results indicated that pSCA1 encoding E7/BCL-xL fusion protein delayed cell death in the pSCA1-transfected dendritic cell line and generated significantly higher E7-specific CD8(+) T-cell-mediated immune responses and better antitumor effects than pSCA1 encoding wild-type E7 gene in vaccinated mice. The antiapoptotic function of BCL-xL is important for the enhancement of antigen-specific CD8(+) T-cell responses in vaccinated mice, because a point mutant of BCL-xL lacking antiapoptotic function was ineffective. These results suggest that strategies to delay suicidal DNA-induced cell death using antiapoptotic proteins may greatly enhance the potency of suicidal DNA. PMID- 14737095 TI - Rapamycin control of exocrine protein levels in saliva after adenoviral vector mediated gene transfer. AB - Transgene-encoded therapeutic secretory proteins can be efficiently secreted from salivary glands into saliva or the bloodstream after adenoviral (Ad)-mediated gene transfer. Since transgene expression from conventional vectors is typically unregulated, we evaluated the rapamycin-based dimerizer regulation system for control of transgene expression in, and consequent exocrine protein secreted from, rat salivary glands. We used human growth hormone (hGH) as a surrogate exocrine secretory protein. Two Ad vectors, Ad C4ZF3, encoding activation and DNA binding domain fusion polypeptides, and Ad Z12-I-GH-2, encoding hGH, were constructed and shown useful in vitro. Thereafter, both vectors were delivered into submandibular glands by retroductal infusion. After 24 h, rapamycin (0, 1, 3 or 10 mg/kg) was administered, and 20 h later hGH levels in saliva were determined. Salivary hGH levels were rapamycin concentration dependent. At a rapamycin dose of 10 mg/kg, total salivary hGH was 693+/-197 ng and the hGH concentration in saliva was 4.6+/-1.3 microg/ml. Over a 16-day experimental period, three separate administrations of rapamycin (3 mg/kg) induced distinct elevations of salivary hGH (approximately 100-200 ng total hGH) that were entirely rapamycin dependent. This study demonstrates for the first time pharmacological control of transgenic exocrine protein production and presence in saliva after salivary gland gene transfer, and the potential for its application to the management of oral, oropharyngeal and upper gastrointestinal tract disorders. PMID- 14737096 TI - Genetic regulation of immune responses to vaccines in early life. AB - Infant immunization is the most cost-effective strategy to prevent infectious diseases in childhood, but is limited by immaturity of the immune system. To define strategies to improve vaccine immunogenicity in early life, the role of genetic and environmental factors in the control of vaccine responses in infant twins was studied. Immune responses to BCG, polio, hepatitis B, diphtheria, pertussis and tetanus vaccines were measured at 5 months of age in 207 Gambian twin pairs recruited at birth. Intrapair correlations for monozygous and dizygous pairs were compared to estimate the environmental and genetic components of variation in responses. High heritability was observed for antibody (Ab) responses to hepatitis B (77%), oral polio (60%), tetanus (44%) and diphtheria (49%) vaccines. Significant heritability was also observed for interferon-gamma and interleukin-13 responses to tetanus, pertussis and some BCG vaccine antigens (39-65%). Non-HLA genes played a dominant role in responses to Ab-inducing vaccines, whereas responses to BCG were predominantly controlled by genes within the HLA class II locus. Genetic factors, particularly non-HLA genes, significantly modulate immune responses to infant vaccination. The identification of the specific genes involved will provide new targets for the development of vaccines and adjuvants for young infants that work independently of HLA. PMID- 14737097 TI - Both risk alleles for FcgammaRIIA and FcgammaRIIIA are susceptibility factors for SLE: a unifying hypothesis. AB - The aim of this study was to analyze in families with SLE for the presence of linkage and the structure and transmission of haplotypes containing alleles for the low-affinity Fcgamma receptors. The Fcgamma receptor polymorphisms FcgammaRIIA-131R/H, FcgammaRIIIA-176F/V and FcgammaRIIIB-NA1/2 and a polymorphism in the FcgammaRIIB gene were genotyped with RFLP, allele-specific PCR or pyrosequencing. Individual SNPs and haplotypes were tested for linkage in multicase families and for association using contingency tables, transmission disequilibrium test and affected family-based control groups in Swedish and Mexican single-case families. No linkage or association could be detected using the FcgammaR polymorphisms in the multicase families. However, an association was found for both FcgammaRIIA-131R and IIIA-176F alleles in the single-case families, but not for IIIB or IIB. Allelic association to SLE was found for a haplotype that included both risk alleles, but not in haplotypes where only one or the other was present. We propose that FcgammaRIIA-131R and FcgammaRIIIA-176F are both risk alleles for SLE transmitted primarily, but not exclusively on a single major haplotype that behaves functionally in a situation similar to that of compound heterozygozity. PMID- 14737098 TI - p53 protein variants: structural and functional similarities with p63 and p73 isoforms. AB - Since its discovery in 1979, many studies have reported that the p53 tumour suppressor protein could be expressed in the form of products smaller than those predicted by the full-length amino-acid sequence. These products differ from full length p53 in their N- or C-terminal regions, but generally conserve the central, DNA-binding domain. They appear to be expressed at rather low levels and to be restricted to particular cell types and/or physiological circumstances, suggesting that they play very narrow and specific roles. Several mechanisms have been proposed to explain their timely occurrence, including alternative splicing, internal initiation of translation or proteolytic cleavage. A precise assessment of the various 'p53 isoforms' reveals striking similarities with several isoforms of the p53 homologous proteins p63 or p73, suggesting that regulated production of specific, N- or C-terminal variants may be a 'trademark' of all family members. In this review, we summarize the published evidence on the structure, mode of production, expression and function of the p53 isoforms, and discuss their properties in the light of recent data on the structure and function of p63/p73 isoforms. PMID- 14737099 TI - Manipulation of nonsense mediated decay identifies gene mutations in colon cancer Cells with microsatellite instability. AB - Cancer cells showing microsatellite instability (MSI) demonstrate a high frequency of acquired frameshift mutations that result in the generation of nonsense mutations. RNA transcripts carrying these nonsense mutations are usually targeted for degradation through the nonsense mediated decay (NMD) pathway. Blocking this pathway with drugs such as emitine, results in the 'stabilization' of these mutant transcripts, which can now be detected on cDNA arrays. Unfortunately, emetine also induces a stress response that results in upregulation of additional transcripts which contribute to the analysis of the array. As a result, identifying which genes truly carry nonsense mutations is made more difficult. To overcome this, we have combined the emetine treatment with actinomycin D, which effectively prevents the upregulation of stress response genes while still stabilizing mutant transcripts. When we applied this modified approach to the analysis of MSI-positive colon cancer cells, we identified mutations in the UVRAG and p300 genes. PMID- 14737100 TI - Truncated ErbB2 receptor (p95ErbB2) is regulated by heregulin through heterodimer formation with ErbB3 yet remains sensitive to the dual EGFR/ErbB2 kinase inhibitor GW572016. AB - The expression of the NH2 terminally truncated ErbB2 receptor (p95ErbB2) in breast cancer correlates with metastatic disease progression compared with the expression of full-length p185ErbB2. We now show that heregulin (HRG), but not EGF, stimulates p95ErbB2 phosphorylation in BT474 breast cancer cells. Furthermore, phospho-p95ErbB2 forms heterodimers with ErbB3, but not EGFR, while p185ErbB2 heterodimerizes with both EGFR and ErbB3. The predilection of p95ErbB2 to heterodimerize with ErbB3 provides an explanation for its regulation by HRG, an ErbB3 ligand. GW572016, a reversible small molecule inhibitor of EGFR and ErbB2 tyrosine kinases, inhibits baseline p95ErbB2 phosphorylation in BT474 cells and tumor xenografts. Inhibition of p95ErbB2, p185ErbB2, and EGFR phosphorylation by GW572016 resulted in the inhibition of downstream phospho-Erk1/2, phospho-AKT, and cyclin D steady-state protein levels. Increased phosphorylation of p95ErbB2 and AKT in response to HRG was abrogated to varying degrees by GW572016. In contrast, trastuzumab did not inhibit p95ErbB2 phosphorylation or the expression of downstream phospho-Erk1/2, phospho-AKT, or cyclin D. It is tempting to speculate that trastuzumab resistance may be mediated in part by the selection of p95ErbB2-expressing breast cancer cells capable of exerting potent growth and prosurvival signals through p95ErbB2-ErbB3 heterodimers. Thus, p95ErbB2 represents a target for therapeutic intervention, and one that is sensitive to GW572016 therapy. PMID- 14737101 TI - Identification of IGFBP-6 as a significantly downregulated gene by beta-catenin in desmoid tumors. AB - Desmoid tumors (aggressive fibromatosis) are locally invasive soft tissue tumors in which beta-catenin-mediated TCF-3-dependent transcription is activated. To provide more insight into the pathophysiology of these tumors, expression profiles were generated using oligonucleotide arrays (Affymetrix). In total, 69 differentially expressed genes were identified in desmoids compared to normal fibroblasts (fascia) from the same patients. The differential expression of a selection of genes was confirmed using RT-PCR and Northern blotting. We further evaluated the insulin-like growth factor-binding protein 6 (IGFBP-6), a gene that was consistently downregulated in all desmoids tested. Promotor studies and electromobility shift assays revealed two functional beta-catenin/TCF-responsive elements in the human IGFBP-6 promoter. These findings suggest that IGFBP-6 is directly downregulated by the beta-catenin/TCF complex in desmoid tumors, and imply a role for the IGF axis in the proliferation of desmoid tumors. PMID- 14737102 TI - Myeloid leukemia with promyelocytic features in transgenic mice expressing hCG NuMA-RARalpha. AB - Acute promyelocytic leukemia (APL) is characterized by the accumulation of abnormal promyelocytes in the bone marrow (BM), and by the presence of a reciprocal chromosomal translocation involving retinoic acid receptor alpha (RARalpha). To date, five RARalpha partner genes have been identified in APL. NuMA-RARalpha was identified in a pediatric case of APL carrying a translocation t(11;17)(q13;q21). Using a construct containing the NuMA-RARalpha fusion gene driven by the human cathepsin G promoter (hCG-NuMA-RARalpha), two transgenic mouse lines were generated. Transgenic mice were observed to have a genetic myeloproliferation (increased granulopoiesis in BM) at an early age, and rapidly developed a myeloproliferative disease-like myeloid leukemia. This leukemia was morphologically and immunophenotypically indistinguishable from human APL, with a penetrance of 100%. The phenotype of transgenic mice was consistent with a blockade of neutrophil differentiation. NuMA-RARalpha is therefore sufficient for disease development in this APL model. PMID- 14737103 TI - p53 deficiency provokes urothelial proliferation and synergizes with activated Ha ras in promoting urothelial tumorigenesis. AB - Mutation and deletion of the p53 tumor suppressor gene are arguably the most prevalent among the multiple genetic alterations found in human bladder cancer, but these p53 defects are primarily associated with the advanced diseases, and their roles in bladder tumor initiation and in synergizing with oncogenes in tumor progression have yet to be defined. Using the mouse uroplakin II gene promoter, we have targeted into urothelium of transgenic mice a dominant-negative mutant of p53 that lacks the DNA-binding domain but retains the tetramerization domain. Urothelium-expressed p53 mutant binds to and stabilizes the endogenous wild-type p53, induces nuclear abnormality, hyperplasia and occasionally dysplasia, without eliciting frank carcinomas. Concurrent expression of the p53 mutant with an activated Ha-ras, the latter of which alone induces urothelial hyperplasia, fails to accelerate tumor formation. In contrast, the expression of the activated Ha-ras in the absence of p53, as accomplished by crossing the activated Ha-ras transgenic mice with the p53 knockout mice, results in early onset bladder tumors that are either low-grade superficial papillary or high grade in nature. These results provide the first in vivo experimental evidence that p53 deficiency predisposes the urothelium to hyperproliferation, but is insufficient for bladder tumorigenesis; that the mere reduction of p53 dosage, as produced in transgenic mice expressing the dominant-negative p53 or in heterozygous p53 knockouts, is incapable of synergizing with Ha-ras to induce bladder tumors; and that the complete loss of p53 is a prerequisite for collaborating with activated Ha-ras to promote bladder tumorigenesis. PMID- 14737104 TI - Downregulation of Muc1 in MMTV-c-Neu tumors. AB - MUC1 is a tumor antigen, overexpressed in approximately 90% of human breast cancers. In normal glandular epithelia, MUC1 is expressed at the apical surface; however, in carcinomas an aberrantly glycosylated form of MUC1 is upregulated and expressed around the entire surface of the cell. Previously, we have shown that a lack of Muc1 significantly delays tumor progression and/or onset in MMTV-PyV-mT and MMTV-Wnt-1 transgenic mice. Here we show that, unlike the models mentioned above, a loss of Muc1 in MMTV-c-Neu mice (MMTV-c-Neu/Muc1(-/-)) altered neither mammary tumor onset nor progression. Moreover, characterization of MMTV-c Neu/Muc1(+/+) tumors revealed that Muc1 expression was repressed at the level of transcription. In contrast, normal mammary gland tissue adjacent to tumor tissue expressed Muc1 and pregnant mammary glands from c-Neu transgenic animals expressed high levels of Muc1. We found that transient transfection of activated ErbB2 into human embryonic kidney 293/MUC1 cells resulted in the repression of MUC1 expression. Further, transient transfection of activated ErbB2 resulted in the inhibition of Muc1 transcriptional activation in luciferase reporter assays. These data suggest that the activation of ErbB2, which only occurs in c-Neu tumors, selectively inhibits Muc1 expression. PMID- 14737105 TI - Phosphatidylinositol-3-OH kinase/AKT and survivin pathways as critical targets for geranylgeranyltransferase I inhibitor-induced apoptosis. AB - Geranylgeranyltransferase I inhibitors (GGTIs) represent a new class of anticancer drugs. However, the mechanism by which GGTIs inhibit tumor cell growth is still unclear. Here, we demonstrate that GGTI-298 and GGTI-2166 induce apoptosis in both cisplatin-sensitive and -resistant human ovarian epithelial cancer cells by inhibition of PI3K/AKT and survivin pathways. Following GGTI-298 or GGTI-2166 treatment, kinase levels of PI3K and AKT were decreased and survivin expression was significantly reduced. Ectopic expression of constitutively active AKT2 and/or survivin significantly rescue human cancer cells from GGTI-298 induced apoptosis. Previous studies have shown that Akt mediates growth factor induced survivin, whereas p53 inhibits survivin expression. However, constitutively active AKT2 failed to rescue the GGTIs downregulation of survivin. Further, GGTIs suppress survivin expression and induce programmed cell death in both wild-type p53 and p53-deficient ovarian cancer cell lines. These data indicate that GGTI-298 and GGTI-2166 induce apoptosis by targeting PI3K/AKT and survivin parallel pathways independent of p53. Owing to the fact that upregulation of Akt and survivin as well as inactivation of p53 are frequently associated with chemoresistance, GGTIs could be valuable agents to overcome antitumor drug resistance. PMID- 14737106 TI - C/EBPalphap30, a myeloid leukemia oncoprotein, limits G-CSF receptor expression but not terminal granulopoiesis via site-selective inhibition of C/EBP DNA binding. AB - Heterozygous mutations of the CEBPA gene are present in 5% of acute myeloid leukemia (AML) cases and often lead to the expression of an N-terminally truncated, 30 kDa isoform, C/EBPalphap30, from an internal translation start site. We have assessed the effect of C/EBPalphap30 on granulopoiesis utilizing C/EBPalphap30-ER, containing the estradiol receptor ligand-binding domain. In contrast to C/EBPalpha-ER, C/EBPalphap30-ER did not induce 32Dcl3 myeloid cell differentiation in IL-3. However, both isoforms, when expressed at high levels, were capable of inhibiting E2F activity in 32Dcl3 cells and of slowing their G1 to S progression. C/EBPalphap30 repressed expression of the endogenous G-CSF receptor several-fold. To facilitate investigation of the effect of C/EBPalphap30 ER on granulopoiesis downstream of G-CSF signalling, we coexpressed exogenous G CSF receptor. C/EBPalphap30-ER/GR cells expressed several granulocytic markers in G-CSF and demonstrated nuclear maturation. Rat C/EBPalpha-ER and C/EBPalphap30 ER, expressed in 293T cells, bound the C/EBP site from the NE gene with similar affinity, as did human C/EBPalpha and C/EBPalphap30. In contrast, C/EBPalphap30 bound the C/EBP sites in the PU.1 or GR gene with 3-6-fold reduced affinity. Thus, the selective inhibition of GR expression by C/EBPalphap30-ER is due in part to its variable affinity for C/EBP sites. Variation in affinity for selected cis elements among isoforms may affect the biology of basic region-leucine zipper (bZIP) proteins. PMID- 14737107 TI - STAT3 and MITF cooperatively induce cellular transformation through upregulation of c-fos expression. AB - The signal transducer and activator of transcription (STAT) family proteins are transcription factors critical in mediating cytokine signaling. Among them, STAT3 is frequently activated in a number of human cancers and transformed cell lines and is implicated in tumorigenesis. However, although constitutively activated STAT3 mutant (STAT3C) leads to cellular transformation, its transformation potential such as colony-forming activity in soft-agar is much weaker than that of v-src. To identify tumorigenic factors that cooperatively induce cellular transformation with STAT3C, we screened the retroviral cDNA library. We found that the microphthalmia-associated transcription factor (MITF), an essential transcription factor for melanocyte development and pigmentation, induces anchorage-independent growth of NIH-3T3 cells in cooperation with STAT3C. Microarray analysis revealed that c-fos is highly expressed in transformants expressing STAT3C and MITF. Promoter analysis and chromatin immunoprecipitation assay suggested that both STAT3 and MITF can cooperatively upregulate the c-fos gene. In addition, the transformation of NIH-3T3 cells by both MITF and STAT3C was significantly suppressed by a dominant-negative AP-1 retrovirus. These data indicate that MITF and STAT3 cooperatively induce c-fos, resulting in cellular transformation. PMID- 14737108 TI - The absence of p53 is critical for the induction of apoptosis by 5-aza-2' deoxycytidine. AB - The absence of functional p53 has complex consequences on the cellular responses to cytotoxic drugs. Here, we have examined the role of p53 in the response to 5 aza-2'-deoxycytidine (5-aza-dC or decitabine). Primary mouse embryonic fibroblasts deficient for p53 undergo apoptosis after treatment with 5-aza-dC. When compared with other demethylating drugs or chemotherapeutic treatments, 5 aza-dC showed the highest selectivity ratio for triggering apoptosis in p53 deficient cells relative to wild-type cells. Moreover, the apoptotic efficacy of 5-aza-dC is proprietary of p53-deficient cells, not being observed in cells lacking other cell-cycle regulators, such as p19ARF, p16INK4a, p21(CIP1/WAF1), E2F-1, or E2F-2. Interestingly, treatment with 5-aza-dC results in the same degree of global genomic hypomethylation in wild-type and p53-null cells. However, wild-type cells activate p53 and arrest at G2/M, whereas p53-null cells accumulate severe chromosomal aberrations and undergo apoptosis. Significantly, the impact of p53-deficiency on the response to 5-aza-dC is not exclusive of primary non-neoplastic cells, but it is also present in neoplastically transformed cells. Finally, treatment of mice bearing genetically defined tumors with nontoxic doses of 5-aza-dC results in therapeutical responses only on tumors lacking p53, but not on tumors lacking p19ARF. Together, our results put forward the hypothesis that the absence of p53 may determine a higher chemotherapeutic index for 5-aza-dC. PMID- 14737109 TI - A novel time-course cDNA microarray analysis method identifies genes associated with the development of cisplatin resistance. AB - In recent years, most cDNA microarray studies of chemotherapeutic drug resistance have not considered the temporal pattern of gene expression. The objective of this study was to examine systematically changes in gene expression of NCI-H226 and NCI-H2170 lung cancer cells treated weekly with IC10 doses of cisplatin. NCI H226 lung cancer cells were treated weekly with an IC10 dose of cisplatin. Candidate genes with a fold change of 2.0 or more were identified from this study. A second experiment was conducted by exposing NCI-H2170 cells to cisplatin doses that were increased in week 4 and decreased in week 5. Overall, 44 genes were differentially expressed in both the NCI-H226 and NCI-H2170 cell lines. In the NCI-H2170 cell line, 24 genes had a twofold gene expression change from weeks 3 to 4. Real-time PCR found a significant correlation of the gene expression changes for seven genes of interest. This small time-ordered series identified novel genes associated with cisplatin resistance. This kind of analysis should be viewed as a first step towards building gene-regulatory networks. PMID- 14737111 TI - The MAP kinase pathway is required for entry into mitosis and cell survival. AB - In this communication, we examined the role of the MAP kinase pathway in the G2/M phase of the cell cycle. Activation of the Plk1 and MAP kinase pathways was initially evaluated in FT210 cells, which arrest at G2 phase at the restrictive temperature (39 degrees C), due to a mutation in the cdc2 gene. Previous studies had shown that these cells enter mitosis at the nonpermissive temperature upon incubation with okadaic acid, a protein phosphatase 1 and 2A inhibitor. We show that treatment of FT210 cells at 39 degrees C with okadaic acid activated Plk1, as shown by hyperphosphorylation and elevated protein kinase activity, and also induced activation of the MAP kinase pathway. The specific Mek inhibitor PD98059 antagonized the okadaic acid-induced activation of both Plk1 and MAP kinases. This suggests that activation of the MAP kinase pathway may contribute to the okadaic acid-induced activation of Plk1 in FT210 cells at 39 degrees C. We also found that PD98059 strongly attenuated progression of HeLa cells through mitosis, and active Mek colocalizes with Plk1 at mitotic structures. To study the potential function of the MAP kinase pathway during mitosis, RNAi was used to specifically deplete five members of this pathway (Raf1, Mek1/2, Erk1/2). Each of these five protein kinases is required for cell proliferation and survival, and depletion of any of these proteins eventually leads to apoptosis. Treatment with Mek inhibitors also inhibited cell proliferation and caused apoptosis. A dramatic increase of Plk1 activities and a moderate increase of Cdc2 activities in Raf1 depleted cells indicate that Raf1-depleted cells arrest in the late G2 or M phase. Mek1 and Erk1 depletion also caused cell cycle arrest at G2, suggesting that these enzymes are required for the G2/M transition, whereas the loss of Mek2 or Erk2 caused arrest at G1. PMID- 14737110 TI - MYCN-mediated regulation of the MRP1 promoter in human neuroblastoma. AB - In the childhood cancer neuroblastoma (NB), the level of expression of the multidrug resistance-associated protein (MRP1) gene is strongly correlated with expression of the MYCN oncogene in primary NB tumors, suggesting that MRP1 may be a target for MYCN-mediated gene regulation. In this study, we show that MYCN induction in human NB cells results in increased MRP1 mRNA and protein levels, which in turn is accompanied by increased drug resistance and enhanced MRP1 mediated drug efflux. Furthermore, luciferase activity from MRP1 promoter/luciferase gene reporter constructs was significantly increased in NB cells with exogenous overexpression of MYCN, whereas activity was decreased in NB cells stably transfected with MYCN-antisense vectors. Decreased luciferase activity was observed with promoter constructs that lacked one or two E-box sequences or had E-box double point mutations, while a truncated MRP1 promoter lacking all three E-boxes exhibited only basal levels of activity. Specific electrophoretic mobility shifts of MRP1 E-box sequences were detected with nuclear extracts from NB cells with MYCN overexpression, and complex formation was inhibited with the addition of antibodies directed against MYCN or MYC. These findings indicate that by interacting with E-box elements within the promoter, MYCN can upregulate MRP1 expression and modulate drug resistance in NB. PMID- 14737112 TI - High-mobility group A1a protein regulates Ras/ERK signaling in MCF-7 human breast cancer cells. AB - High-mobility group (HMG) A1 proteins are gene regulatory factors whose overexpression is frequently observed in naturally occurring human cancers. The overexpression of transgenic HMGA1 proteins in cells results in neoplastic transformation and promotes progression to malignant cellular phenotypes. To understand the underlying molecular and biological events involved in these phenomena, we used oligonucleotide microarray analyses to generate an HMGA1a induced expression profile for approximately 22,000 genes. This gene expression profile was generated using a well-characterized transgenic human MCF-7 mammary adenocarcinoma cell line in which overexpression of transgenic HMGA1 promotes a transition to a more malignant and metastatic phenotype. Microarray expression analyses, together with independent quantitative real-time reverse transcriptase polymerase chain reaction results, indicate that HMGA1a regulates genes involved in the Ras-extracellular signal-related kinase (Ras/ERK) mitogenic signaling pathway, including KIT ligand and caveolins 1 and 2. We also found that many cholesterol biosynthesis genes were decreased in cells overexpressing HMGA1a. Cholesterol depletion, decreased caveolin, and increased KIT ligand expression, are all independently associated with the activation of Ras/ERK signaling. Upon further analysis, we found that sensitivity to epidermal growth factor activation of ERK phosphorylation was significantly higher, and that cholesterol was significantly depleted, in cells overexpressing HMGA1a. The cumulative evidence indicates that one likely mechanism by which the HMGA1a protein promotes malignant changes in cells is through increased sensitivity to the activation of the Ras/ERK signaling pathway. PMID- 14737113 TI - PTEN inhibits cell proliferation and induces apoptosis by downregulating cell surface IGF-IR expression in prostate cancer cells. AB - PTEN is a tumor suppressor gene that is frequently mutated in human tumors. It functions primarily as a lipid phosphatase and plays a key role in the regulation of phosphatidylinositol-3'-kinase. PTEN appears to play a crucial role in modulating apoptosis by reducing the levels of PtdIns(3,4,5)P3, a phospholipid that activates AKT, a central regulator of apoptosis. To understand the role of PTEN in regulating cell proliferation and apoptosis, we stably overexpressed PTEN in PC3 cells, which are prostate cancer cells that lack PTEN. Overexpression of PTEN in two different clones inhibited cell proliferation and increased serum starvation-induced apoptosis, as compared to control cells. Interestingly, PTEN overexpression resulted in a 44-60% reduction in total insulin-like growth factor I receptor (IGF-IR) protein levels and a 49-64% reduction in cell surface IGF-IR expression. [35S]methionine pulse experiments in PC3 cells overexpressing PTEN demonstrated that these cells synthesize significantly lower levels of the IGF-IR precursor, whereas PTEN overexpression had no effect on IGF-IR degradation. Taken together, our results show that PTEN can regulate cell proliferation and apoptosis through inhibition of IGF-IR synthesis. These results have important implications for understanding the roles of PTEN and the IGF-IR in prostate cancer cell tumorigenesis. PMID- 14737114 TI - Gene expression analysis reveals evidence for inactivation of the TGF-beta signaling cascade in autonomously functioning thyroid nodules. AB - Molecular events that lead to the development of autonomously functioning thyroid nodules (AFTNs) are somatic mutations of the thyrotropin receptor (TSHR) in approximately 60% of the nodules and less frequently, somatic mutations in the Gsalpha protein. However, AFTNs without known mutations indicate that other causes remain to be identified. Moreover, the impact of constitutively activating TSHR mutations on the signal transduction network of the thyroid epithelial cell is unknown. We therefore investigated gene expression in 15 AFTNs and their surrounding tissue using Affymetrix GeneChips. Most prominently, data analysis revealed a changed pattern of gene expression in the TGF-beta signaling cascade and 25 differentially regulated genes in AFTNs, including thyroid peroxidase, type I iodothyronine deiodinase and sialyltransferase (SIAT) 1. Strikingly coexpression of SIAT 1 and TSHR in COS-7 cells increased TSH binding and cell surface expression of the TSHR. Moreover, differences in gene expression patterns for AFTNs with and without TSHR mutations indicate specific alterations of signal transduction in AFTNs without TSHR mutations. These results suggest that AFTNs with TSHR mutations harbor further mechanisms of forward stimulation. Furthermore, they give important leads to elucidate the molecular etiology of AFTNs without TSHR mutations. PMID- 14737115 TI - Src-dependent ERK5 and Src/EGFR-dependent ERK1/2 activation is required for cell proliferation by asbestos. AB - Crocidolite asbestos elicits oxidative stress and cell proliferation, but the signaling cascades linked to these outcomes are unclear. To determine the role of mitogen-activated protein kinases (MAPK) in asbestos-induced cell signaling, we evaluated the effects of crocidolite asbestos, EGF and H2O2, on MAPK activation in murine lung epithelial cells (C10 line). In contrast to rapid and transient activation of extracellular signal-regulated kinase 5 (ERK5) by EGF or H2O2, asbestos caused protracted oxidant-dependent ERK5 activation that was inhibited by an Src kinase inhibitor (PP2), but not by an inhibitor of epidermal growth factor receptor (EGFR) phosphorylation (AG1478). ERK1/2 activation by asbestos was inhibited by either PP2 or AG1478. To confirm the involvement of Src in ERK1/2 and ERK5 activation, a dominant-negative Src construct was used. These experiments showed that Src was essential for ERK1/2 and also ERK5 phosphorylation by asbestos. Time frame studies indicated immediate activation of Src by asbestos fibers, whereas EGFR phosphorylation occurred subsequently. Data suggest that asbestos causes activation of ERK5 through an EGFR-independent pathway, whereas ERK1/2 activation is dependent on Src through a mechanism involving phosphorylation of the EGFR. Furthermore, Src, ERK1/2 and ERK5 activation are essential for cell proliferation by asbestos. The use of a dominant-negative ERK5 construct caused selective downregulation of c-jun expression, whereas inhibition of Src by PP2 or MEK1 by PD98059 caused decreases in c-fos, fra-1 and c-jun expression in asbestos-exposed C10 cells. These observations may have broad relevance to cell proliferation by carcinogenic mineral fibers and oxidants. PMID- 14737116 TI - Differences in stability of repressor complexes at promoters underlie distinct roles for Rb family members. AB - Oncogenic transformation of cells can induce the cyclin-dependent kinase inhibitor, p16, which leads to hypophosphorylation and activation of retinoblastoma (Rb). Rb is capable of causing permanent growth arrest, which may underlie its role as a tumor suppressor. We show that repression by Rb at E2F target gene promoters involves the establishment of a stable repressor complex that is not displaced by the overexpression of E2F-1. Rather than displacing Rb, excess E2F-1 instead recruits more Rb, leading to direct transcriptional repression. In contrast, the Rb family members, p130 and p107, which have not been demonstrated to be tumor suppressors, bind preferentially to target promoters in the absence of growth factors and in proliferating cells, respectively, and these repressor complexes are displaceable by E2F-1. Heterochromatin protein 1 (HP1), which interacts with Rb, is associated with these distinct repressor complexes and follows a similar pattern of stability/displaceability. Efficient growth arrest by p16/Rb is dependent on histone H3 lysine 9 methylation, which provides a binding site for HP1. We propose that these differences in the stability of repressor complexes at promoters may, in part, underlie the different roles of Rb vs p130 and p107 in cell cycle regulation and tumor suppression. PMID- 14737117 TI - Caffeine inhibits homology-directed repair of I-SceI-induced DNA double-strand breaks. AB - We recently reported that two Chinese hamster mutants deficient in the RAD51 paralogs XRCC2 and XRCC3 show reduced radiosensitization after treatment with caffeine, thus implicating homology-directed repair (HDR) of DNA double-strand breaks (DSBs) in the mechanism of caffeine radiosensitization. Here, we investigate directly the effect of caffeine on HDR initiated by DSBs induced by a rare cutting endonuclease (I-SceI) into one of two direct DNA repeats. The results demonstrate a strong inhibition by caffeine of HDR in wild-type cells, and a substantial reduction of this effect in HDR-deficient XRCC3 mutant cells. Inhibition of HDR and cell radiosensitization to killing shows similar dependence on caffeine concentration suggesting a cause-effect relationship between these effects. UCN-01, a kinase inhibitor that effectively abrogates checkpoint activation in irradiated cells, has only a small effect on HDR, indicating that similar to radiosensitization, inhibition of checkpoint signaling is not sufficient for HDR inhibition. Recombination events occurring during treatment with caffeine are characterized by rearrangements reminiscent to those previously reported for the XRCC3 mutant, and immunofluorescence microscopy demonstrates significantly reduced formation of IR-specific RAD51 foci after caffeine treatment. In summary, our results identify inhibition of HDR as a significant contributor to caffeine radiosensitization. PMID- 14737118 TI - Divergence of Genbank and human tumor Bcl-2 sequences and implications for binding affinity to key apoptotic proteins. AB - Heterodimerization of antiapoptotic and pro-apoptotic Bcl-2 family of proteins provides an important mechanism for apoptosis regulation. Knowledge about key amino acids in the binding groove of native Bcl-2 contributing to this interaction will greatly facilitate the design of Bcl-2-specific inhibitors. There are two different Bcl-2 sequences, M13994 and M14745, in Genbank. Chimeric proteins Bcl-2(1) and Bcl-2(2) derived from the above sequences, although similar in structure, showed different binding affinities to Bak and Bad BH3 peptides (Petros et al., 2001). In this study, we show that the Bcl-2(1) sequence in normal and tumor human tissue samples differs from M13994 and M14745, and contains P59, T96, R110, S117 and G237. The actual sequence in the binding pocket matches the Bcl-2-Ig fusion sequence X06487, originally identified in a t(14:18) translocation of the Bcl-2 gene, associated with follicular lymphoma. The possible effects of the observed amino acid differences compared to M13994 and M14745 were investigated by combining structural data with fluorescence anisotropy. G110R substitution confers on Bcl-2(1) substantially increased binding affinity to Bak, Bad and Bax BH3 peptides, demonstrating that R110 is a key contributor to the BH3 binding affinity of Bcl-2. Although NMR structure did not predict R110 involvement in binding to these BH3 peptides, fluorescence anisotropy data clearly points to a critical role for this residue in binding to pro-apoptotic Bcl-2 family members. PMID- 14737119 TI - Transcriptional repressor BCL-6 immortalizes germinal center-like B cells in the absence of p53 function. AB - Chromosomal rearrangements in non-Hodgkin's B-cell lymphoma implicate BCL-6 as an oncogene, yet direct evidence for BCL-6 acting as an oncogene in B cells has been lacking. Here, we show that BCL-6 can immortalize primary B cells, but only in the absence of p53 tumor suppressor function. The expression of BCL-6 led to greatly increased B-cell proliferation, particularly in response to CD40 stimulation. Furthermore, BCL-6-infected p53-deficient B cells gave rise to immortalized cell lines that could be maintained by CD40 stimulation. We found that in primary mouse B cells, BCL-6 repressed expression of the Blimp-1, p27kip1, and cyclin D2 target genes. BCL-6 did not markedly repress the PDCD2 and BCL-XL target genes. The BCL-6 immortalized cell lines had a phenotype consistent with germinal center B cells, they expressed the germinal center-specific M17 gene, and a significant fraction of the cells stained positive with PNA. Our data indicate that BCL-6 may act to maintain B cells in a germinal center-like state, and repression of Blimp-1 by BCL-6 may be particularly crucial for stabilization of the germinal center phenotype. Our data also suggest that disruption of the p53 pathway may be crucial for the development of BCL-6-expressing B-cell lymphomas. PMID- 14737120 TI - Applying the principles of stem-cell biology to cancer. PMID- 14737121 TI - Hedgehog signalling in cancer formation and maintenance. PMID- 14737122 TI - Intravital imaging of cell movement in tumours. PMID- 14737123 TI - The 14-3-3 cancer connection. PMID- 14737124 TI - Opinion: Searching for the elusive targets of farnesyltransferase inhibitors. PMID- 14737125 TI - Opinion: Comparative biology of mouse versus human cells: modelling human cancer in mice. PMID- 14737126 TI - Towards a German elite. PMID- 14737128 TI - NASA's drive to revisit the Moon leaves no scope for Hubble. PMID- 14737127 TI - Back to the moon. PMID- 14737129 TI - Bird flu sparks worldwide bid to prevent human pandemic. PMID- 14737130 TI - Director quits as cash cutbacks hit Californian labs. PMID- 14737131 TI - Feathers fly over welfare of hemmed-in hens. PMID- 14737132 TI - Ministers prepare to back neuroscience network. PMID- 14737133 TI - Resignation threats add steel to French revolt. PMID- 14737134 TI - Europe's researchers up in arms over clinical-trial rules. PMID- 14737136 TI - Telepathic charm seduces audience at paranormal debate. PMID- 14737135 TI - Health concerns prompt US review of exotic-pet trade. PMID- 14737138 TI - Star quality. PMID- 14737139 TI - Microbiology: gut reaction. PMID- 14737140 TI - Milton and Galileo would back BMJ on free speech. PMID- 14737141 TI - X-ray clues to viability of loop quantum gravity. PMID- 14737142 TI - Return of bone archives is a loss to humanity. PMID- 14737143 TI - The hole truth. PMID- 14737148 TI - A meeting with Enrico Fermi. PMID- 14737149 TI - Astronomy: a problem of distance. PMID- 14737150 TI - Cell division: burning the spindle at both ends. PMID- 14737151 TI - Nanotechnology: dreams of a hollow future. PMID- 14737153 TI - Psychology: insight and the sleep committee. PMID- 14737154 TI - Biogeochemistry: carbon budget in the black. PMID- 14737155 TI - Virology: a class act. PMID- 14737157 TI - Neuroplasticity: changes in grey matter induced by training. PMID- 14737158 TI - Animal behaviour: cognitive bias and affective state. PMID- 14737159 TI - Structure of the dengue virus envelope protein after membrane fusion. AB - Dengue virus enters a host cell when the viral envelope glycoprotein, E, binds to a receptor and responds by conformational rearrangement to the reduced pH of an endosome. The conformational change induces fusion of viral and host-cell membranes. A three-dimensional structure of the soluble E ectodomain (sE) in its trimeric, postfusion state reveals striking differences from the dimeric, prefusion form. The elongated trimer bears three 'fusion loops' at one end, to insert into the host-cell membrane. Their structure allows us to model directly how these fusion loops interact with a lipid bilayer. The protein folds back on itself, directing its carboxy terminus towards the fusion loops. We propose a fusion mechanism driven by essentially irreversible conformational changes in E and facilitated by fusion-loop insertion into the outer bilayer leaflet. Specific features of the folded-back structure suggest strategies for inhibiting flavivirus entry. PMID- 14737160 TI - Conformational change and protein-protein interactions of the fusion protein of Semliki Forest virus. AB - Fusion of biological membranes is mediated by specific lipid-interacting proteins that induce the formation and expansion of an initial fusion pore. Here we report the crystal structure of the ectodomain of the Semliki Forest virus fusion glycoprotein E1 in its low-pH-induced trimeric form. E1 adopts a folded-back conformation that, in the final post-fusion form of the full-length protein, would bring the fusion peptide loop and the transmembrane anchor to the same end of a stable protein rod. The observed conformation of the fusion peptide loop is compatible with interactions only with the outer leaflet of the lipid bilayer. Crystal contacts between fusion peptide loops of adjacent E1 trimers, together with electron microscopy observations, suggest that in an early step of membrane fusion, an intermediate assembly of five trimers creates two opposing nipple-like deformations in the viral and target membranes, leading to formation of the fusion pore. PMID- 14737161 TI - A distance of 133-137 parsecs to the Pleiades star cluster. AB - Nearby 'open' clusters of stars (those that are not gravitationally bound) have played a crucial role in the development of stellar astronomy because, as a consequence of the stars having a common age, they provide excellent natural laboratories to test theoretical stellar models. Clusters also play a fundamental part in determining distance scales. The satellite Hipparcos surprisingly found that an extensively studied open cluster--the Pleiades (also known as the Seven Sisters)--had a distance of D = 118 +/- 4 pc (refs 2, 3), about ten per cent smaller than the accepted value. The discrepancy generated a spirited debate because the implication was that either current stellar models were incorrect by a surprising amount or Hipparcos was giving incorrect distances. Here we report the orbital parameters of the bright double star Atlas in the Pleiades, using long-baseline optical/infrared interferometry. From the data we derive a firm lower bound of D > 127 pc, with the most likely range being 133 < D < 137 pc. Our result reaffirms the fidelity of current stellar models. PMID- 14737162 TI - The microscopic nature of localization in the quantum Hall effect. AB - The quantum Hall effect arises from the interplay between localized and extended states that form when electrons, confined to two dimensions, are subject to a perpendicular magnetic field. The effect involves exact quantization of all the electronic transport properties owing to particle localization. In the conventional theory of the quantum Hall effect, strong-field localization is associated with a single-particle drift motion of electrons along contours of constant disorder potential. Transport experiments that probe the extended states in the transition regions between quantum Hall phases have been used to test both the theory and its implications for quantum Hall phase transitions. Although several experiments on highly disordered samples have affirmed the validity of the single-particle picture, other experiments and some recent theories have found deviations from the predicted universal behaviour. Here we use a scanning single-electron transistor to probe the individual localized states, which we find to be strikingly different from the predictions of single-particle theory. The states are mainly determined by Coulomb interactions, and appear only when quantization of kinetic energy limits the screening ability of electrons. We conclude that the quantum Hall effect has a greater diversity of regimes and phase transitions than predicted by the single-particle framework. Our experiments suggest a unified picture of localization in which the single particle model is valid only in the limit of strong disorder. PMID- 14737163 TI - Reburial of fossil organic carbon in marine sediments. AB - Marine sediments act as the ultimate sink for organic carbon, sequestering otherwise rapidly cycling carbon for geologic timescales. Sedimentary organic carbon burial appears to be controlled by oxygen exposure time in situ, and much research has focused on understanding the mechanisms of preservation of organic carbon. In this context, combustion-derived black carbon has received attention as a form of refractory organic carbon that may be preferentially preserved in soils and sediments. However, little is understood about the environmental roles, transport and distribution of black carbon. Here we apply isotopic analyses to graphitic black carbon samples isolated from pre-industrial marine and terrestrial sediments. We find that this material is terrestrially derived and almost entirely depleted of radiocarbon, suggesting that it is graphite weathered from rocks, rather than a combustion product. The widespread presence of fossil graphitic black carbon in sediments has therefore probably led to significant overestimates of burial of combustion-derived black carbon in marine sediments. It could be responsible for biasing radiocarbon dating of sedimentary organic carbon, and also reveals a closed loop in the carbon cycle. Depending on its susceptibility to oxidation, this recycled carbon may be locked away from the biologically mediated carbon cycle for many geologic cycles. PMID- 14737164 TI - Melting of iron at the physical conditions of the Earth's core. AB - Seismological data can yield physical properties of the Earth's core, such as its size and seismic anisotropy. A well-constrained iron phase diagram, however, is essential to determine the temperatures at core boundaries and the crystal structure of the solid inner core. To date, the iron phase diagram at high pressure has been investigated experimentally through both laser-heated diamond anvil cell and shock-compression techniques, as well as through theoretical calculations. Despite these contributions, a consensus on the melt line or the high-pressure, high-temperature phase of iron is lacking. Here we report new and re-analysed sound velocity measurements of shock-compressed iron at Earth-core conditions. We show that melting starts at 225 +/- 3 GPa (5,100 +/- 500 K) and is complete at 260 +/- 3 GPa (6,100 +/- 500 K), both on the Hugoniot curve-the locus of shock-compressed states. This new melting pressure is lower than previously reported, and we find no evidence for a previously reported solid-solid phase transition on the Hugoniot curve near 200 GPa (ref. 16). PMID- 14737165 TI - Chicken welfare is influenced more by housing conditions than by stocking density. AB - Intensive broiler (meat) chicken production now exceeds 800 million birds each year in the United Kingdom and 2 x 10(10) birds worldwide, but it attracts accusations of poor welfare. The European Union is currently adopting standards for broilers aimed at a chief welfare concern--namely, overcrowding--by limiting maximum 'stocking density' (bird weight per unit area). It is not clear, however, whether this will genuinely improve bird welfare because evidence is contradictory. Here we report on broiler welfare in relation to the European Union proposals through a large-scale study (2.7 million birds) with the unprecedented cooperation of ten major broiler producers in an experimental manipulation of stocking density under a range of commercial conditions. Producer companies stocked birds to five different final densities, but otherwise followed company practice, which we recorded in addition to temperature, humidity, litter and air quality. We assessed welfare through mortality, physiology, behaviour and health, with an emphasis on leg health and walking ability. Our results show that differences among producers in the environment that they provide for chickens have more impact on welfare than has stocking density itself. PMID- 14737166 TI - Travelling waves in the occurrence of dengue haemorrhagic fever in Thailand. AB - Dengue fever is a mosquito-borne virus that infects 50-100 million people each year. Of these infections, 200,000-500,000 occur as the severe, life-threatening form of the disease, dengue haemorrhagic fever (DHF). Large, unanticipated epidemics of DHF often overwhelm health systems. An understanding of the spatial temporal pattern of DHF incidence would aid the allocation of resources to combat these epidemics. Here we examine the spatial-temporal dynamics of DHF incidence in a data set describing 850,000 infections occurring in 72 provinces of Thailand during the period 1983 to 1997. We use the method of empirical mode decomposition to show the existence of a spatial-temporal travelling wave in the incidence of DHF. We observe this wave in a three-year periodic component of variance, which is thought to reflect host-pathogen population dynamics. The wave emanates from Bangkok, the largest city in Thailand, moving radially at a speed of 148 km per month. This finding provides an important starting point for detecting and characterizing the key processes that contribute to the spatial-temporal dynamics of DHF in Thailand. PMID- 14737167 TI - A primitive Y chromosome in papaya marks incipient sex chromosome evolution. AB - Many diverse systems for sex determination have evolved in plants and animals. One involves physically distinct (heteromorphic) sex chromosomes (X and Y, or Z and W) that are homozygous in one sex (usually female) and heterozygous in the other (usually male). Sex chromosome evolution is thought to involve suppression of recombination around the sex determination genes, rendering permanently heterozygous a chromosomal region that may then accumulate deleterious recessive mutations by Muller's ratchet, and fix deleterious mutations by hitchhiking as nearby favourable mutations are selected on the Y chromosome. Over time, these processes may cause the Y chromosome to degenerate and to diverge from the X chromosome over much of its length; for example, only 5% of the human Y chromosome still shows X-Y recombination. Here we show that papaya contains a primitive Y chromosome, with a male-specific region that accounts for only about 10% of the chromosome but has undergone severe recombination suppression and DNA sequence degeneration. This finding provides direct evidence for the origin of sex chromosomes from autosomes. PMID- 14737168 TI - Sleep inspires insight. AB - Insight denotes a mental restructuring that leads to a sudden gain of explicit knowledge allowing qualitatively changed behaviour. Anecdotal reports on scientific discovery suggest that pivotal insights can be gained through sleep. Sleep consolidates recent memories and, concomitantly, could allow insight by changing their representational structure. Here we show a facilitating role of sleep in a process of insight. Subjects performed a cognitive task requiring the learning of stimulus-response sequences, in which they improved gradually by increasing response speed across task blocks. However, they could also improve abruptly after gaining insight into a hidden abstract rule underlying all sequences. Initial training establishing a task representation was followed by 8 h of nocturnal sleep, nocturnal wakefulness, or daytime wakefulness. At subsequent retesting, more than twice as many subjects gained insight into the hidden rule after sleep as after wakefulness, regardless of time of day. Sleep did not enhance insight in the absence of initial training. A characteristic antecedent of sleep-related insight was revealed in a slowing of reaction times across sleep. We conclude that sleep, by restructuring new memory representations, facilitates extraction of explicit knowledge and insightful behaviour. PMID- 14737169 TI - Lymphocyte egress from thymus and peripheral lymphoid organs is dependent on S1P receptor 1. AB - Adaptive immunity depends on T-cell exit from the thymus and T and B cells travelling between secondary lymphoid organs to survey for antigens. After activation in lymphoid organs, T cells must again return to circulation to reach sites of infection; however, the mechanisms regulating lymphoid organ exit are unknown. An immunosuppressant drug, FTY720, inhibits lymphocyte emigration from lymphoid organs, and phosphorylated FTY720 binds and activates four of the five known sphingosine-1-phosphate (S1P) receptors. However, the role of S1P receptors in normal immune cell trafficking is unclear. Here we show that in mice whose haematopoietic cells lack a single S1P receptor (S1P1; also known as Edg1) there are no T cells in the periphery because mature T cells are unable to exit the thymus. Although B cells are present in peripheral lymphoid organs, they are severely deficient in blood and lymph. Adoptive cell transfer experiments establish an intrinsic requirement for S1P1 in T and B cells for lymphoid organ egress. Furthermore, S1P1-dependent chemotactic responsiveness is strongly upregulated in T-cell development before exit from the thymus, whereas S1P1 is downregulated during peripheral lymphocyte activation, and this is associated with retention in lymphoid organs. We find that FTY720 treatment downregulates S1P1, creating a temporary pharmacological S1P1-null state in lymphocytes, providing an explanation for the mechanism of FTY720-induced lymphocyte sequestration. These findings establish that S1P1 is essential for lymphocyte recirculation and that it regulates egress from both thymus and peripheral lymphoid organs. PMID- 14737170 TI - The mitochondrial calcium uniporter is a highly selective ion channel. AB - During intracellular Ca2+ signalling mitochondria accumulate significant amounts of Ca2+ from the cytosol. Mitochondrial Ca2+ uptake controls the rate of energy production, shapes the amplitude and spatio-temporal patterns of intracellular Ca2+ signals, and is instrumental to cell death. This Ca2+ uptake is undertaken by the mitochondrial Ca2+ uniporter (MCU) located in the organelle's inner membrane. The uniporter passes Ca2+ down the electrochemical gradient maintained across this membrane without direct coupling to ATP hydrolysis or transport of other ions. Carriers are characterized by turnover numbers that are typically 1,000-fold lower than ion channels, and until now it has been unclear whether the MCU is a carrier or a channel. By patch-clamping the inner mitochondrial membrane, we identified a previously unknown Ca2+-selective ion channel sensitive to inhibitors of mitochondrial Ca2+ uptake. Our data indicate that this unique channel binds Ca2+ with extremely high affinity (dissociation constant < or =2 nM), enabling high Ca2+ selectivity despite relatively low cytoplasmic Ca2+ concentrations. The channel is inwardly rectifying, making it especially effective for Ca2+ uptake into energized mitochondria. Thus, we conclude that the properties of the current mediated by this novel channel are those of the MCU. PMID- 14737171 TI - The MAPK Hog1 recruits Rpd3 histone deacetylase to activate osmoresponsive genes. AB - Regulation of gene expression by mitogen-activated protein kinases (MAPKs) is essential for proper cell adaptation to extracellular stimuli. Exposure of yeast cells to high osmolarity results in rapid activation of the MAPK Hog1, which coordinates the transcriptional programme required for cell survival on osmostress. The mechanisms by which Hog1 and MAPKs in general regulate gene expression are not completely understood, although Hog1 can modify some transcription factors. Here we propose that Hog1 induces gene expression by a mechanism that involves recruiting a specific histone deacetylase complex to the promoters of genes regulated by osmostress. Cells lacking the Rpd3-Sin3 histone deacetylase complex are sensitive to high osmolarity and show compromised expression of osmostress genes. Hog1 interacts physically with Rpd3 in vivo and in vitro and, on stress, targets the deacetylase to specific osmostress responsive genes. Binding of the Rpd3-Sin3 complex to specific promoters leads to histone deacetylation, entry of RNA polymerase II and induction of gene expression. Together, our data indicate that targeting of the Rpd3 histone deacetylase to osmoresponsive promoters by the MAPK Hog1 is required to induce gene expression on stress. PMID- 14737172 TI - Shifting the balance. PMID- 14737173 TI - Mapping opportunities. PMID- 14737174 TI - Graduate journal: a tough challenge. PMID- 14737175 TI - Scientists and societies: giving young European students a voice. PMID- 14737177 TI - Susceptibility to leprosy is associated with PARK2 and PACRG. AB - Leprosy is caused by Mycobacterium leprae and affects about 700,000 individuals each year. It has long been thought that leprosy has a strong genetic component, and recently we mapped a leprosy susceptibility locus to chromosome 6 region q25 q26 (ref. 3). Here we investigate this region further by using a systematic association scan of the chromosomal interval most likely to harbour this leprosy susceptibility locus. In 197 Vietnamese families we found a significant association between leprosy and 17 markers located in a block of approx. 80 kilobases overlapping the 5' regulatory region shared by the Parkinson's disease gene PARK2 and the co-regulated gene PACRG. Possession of as few as two of the 17 risk alleles was highly predictive of leprosy. This was confirmed in a sample of 975 unrelated leprosy cases and controls from Brazil in whom the same alleles were strongly associated with leprosy. Variants in the regulatory region shared by PARK2 and PACRG therefore act as common risk factors for leprosy. PMID- 14737178 TI - JAK/STAT, Raf/MEK/ERK, PI3K/Akt and BCR-ABL in cell cycle progression and leukemogenesis. AB - The roles of the JAK/STAT, Raf/MEK/ERK and PI3K/Akt signal transduction pathways and the BCR-ABL oncoprotein in leukemogenesis and their importance in the regulation of cell cycle progression and apoptosis are discussed in this review. These pathways have evolved regulatory proteins, which serve to limit their proliferative and antiapoptotic effects. Small molecular weight cell membrane permeable drugs that target these pathways have been developed for leukemia therapy. One such example is imatinib mesylate, which targets the BCR-ABL kinase as well as a few structurally related kinases. This drug has proven to be effective in the treatment of CML patients. However, leukemic cells have evolved mechanisms to become resistant to this drug. A means to combat drug resistance is to target other prominent signaling components involved in the pathway or to inhibit BCR-ABL by other mechanisms. Treatment of imatinib-resistant leukemia cells with drugs that target Ras (farnysyl transferase inhibitors) or with the protein destabilizer geldanamycin has proven to be a means to inhibit the growth of resistant cells. This review will tie together three important signal transduction pathways involved in the regulation of hematopoietic cell growth and indicate how their expression is dysregulated by the BCR-ABL oncoprotein. PMID- 14737181 TI - Science on the rise in developing countries. PMID- 14737182 TI - JAMM: a metalloprotease-like zinc site in the proteasome and signalosome. AB - The JAMM (JAB1/MPN/Mov34 metalloenzyme) motif in Rpn11 and Csn5 underlies isopeptidase activities intrinsic to the proteasome and signalosome, respectively. We show here that the archaebacterial protein AfJAMM possesses the key features of a zinc metalloprotease, yet with a distinct fold. The histidine and aspartic acid of the conserved EX(n)HS/THX(7)SXXD motif coordinate a zinc, whereas the glutamic acid hydrogen-bonds an aqua ligand. By analogy to the active site of thermolysin, we predict that the glutamic acid serves as an acid-base catalyst and the second serine stabilizes a tetrahedral intermediate. Mutagenesis of Csn5 confirms these residues are required for Nedd8 isopeptidase activity. The active site-like architecture specified by the JAMM motif motivates structure based approaches to the study of JAMM domain proteins and the development of therapeutic proteasome and signalosome inhibitors. PMID- 14737183 TI - Dorsoventral patterning of the mouse coat by Tbx15. AB - Many members of the animal kingdom display coat or skin color differences along their dorsoventral axis. To determine the mechanisms that control regional differences in pigmentation, we have studied how a classical mouse mutation, droopy ear (de(H)), affects dorsoventral skin characteristics, especially those under control of the Agouti gene. Mice carrying the Agouti allele black-and-tan (a(t)) normally have a sharp boundary between dorsal black hair and yellow ventral hair; the de(H) mutation raises the pigmentation boundary, producing an apparent dorsal-to-ventral transformation. We identify a 216 kb deletion in de(H) that removes all but the first exon of the Tbx15 gene, whose embryonic expression in developing mesenchyme correlates with pigmentary and skeletal malformations observed in de(H)/de(H) animals. Construction of a targeted allele of Tbx15 confirmed that the de(H) phenotype was caused by Tbx15 loss of function. Early embryonic expression of Tbx15 in dorsal mesenchyme is complementary to Agouti expression in ventral mesenchyme; in the absence of Tbx15, expression of Agouti in both embryos and postnatal animals is displaced dorsally. Transplantation experiments demonstrate that positional identity of the skin with regard to dorsoventral pigmentation differences is acquired by E12.5, which is shortly after early embryonic expression of Tbx15. Fate-mapping studies show that the dorsoventral pigmentation boundary is not in register with a previously identified dermal cell lineage boundary, but rather with the limb dorsoventral boundary. Embryonic expression of Tbx15 in dorsolateral mesenchyme provides an instructional cue required to establish the future positional identity of dorsal dermis. These findings represent a novel role for T-box gene action in embryonic development, identify a previously unappreciated aspect of dorsoventral patterning that is widely represented in furred mammals, and provide insight into the mechanisms that underlie region-specific differences in body morphology. PMID- 14737184 TI - Cell-passage activity is required for the malarial parasite to cross the liver sinusoidal cell layer. AB - Liver infection is an obligatory step in malarial transmission, but it remains unclear how the sporozoites gain access to the hepatocytes, which are separated from the circulatory system by the liver sinusoidal cell layer. We found that a novel microneme protein, named sporozoite microneme protein essential for cell traversal (SPECT), is produced by the liver-infective sporozoite of the rodent malaria parasite, Plasmodium berghei. Targeted disruption of the spect gene greatly reduced sporozoite infectivity to the liver. In vitro cell invasion assays revealed that these disruptants can infect hepatocytes normally but completely lack their cell passage ability. Their apparent liver infectivity was, however, restored by depletion of Kupffer cells, hepatic macrophages included in the sinusoidal cell layer. These results show that malarial sporozoites access hepatocytes through the liver sinusoidal cell layer by cell traversal motility mediated by SPECT and strongly suggest that Kupffer cells are main routes for this passage. Our findings may open the way for novel malaria transmission blocking strategies that target molecules involved in sporozoite migration to the hepatocyte. PMID- 14737186 TI - HIV-1 Nef binds the DOCK2-ELMO1 complex to activate rac and inhibit lymphocyte chemotaxis. AB - The infectious cycle of primate lentiviruses is intimately linked to interactions between cells of the immune system. Nef, a potent virulence factor, alters cellular environments to increase lentiviral replication in the host, yet the mechanisms underlying these effects have remained elusive. Since Nef likely functions as an adaptor protein, we exploited a proteomic approach to directly identify molecules that Nef targets to subvert the signaling machinery in T cells. We purified to near homogeneity a major Nef-associated protein complex from T cells and identified by mass spectroscopy its subunits as DOCK2-ELMO1, a key activator of Rac in antigen- and chemokine-initiated signaling pathways, and Rac. We show that Nef activates Rac in T cell lines and in primary T cells following infection with HIV-1 in the absence of antigenic stimuli. Nef activates Rac by binding the DOCK2-ELMO1 complex, and this interaction is linked to the abilities of Nef to inhibit chemotaxis and promote T cell activation. Our data indicate that Nef targets a critical switch that regulates Rac GTPases downstream of chemokine- and antigen-initiated signaling pathways. This interaction enables Nef to influence multiple aspects of T cell function and thus provides an important mechanism by which Nef impacts pathogenesis by primate lentiviruses. PMID- 14737187 TI - Similarities and differences in genome-wide expression data of six organisms. AB - Comparing genomic properties of different organisms is of fundamental importance in the study of biological and evolutionary principles. Although differences among organisms are often attributed to differential gene expression, genome-wide comparative analysis thus far has been based primarily on genomic sequence information. We present a comparative study of large datasets of expression profiles from six evolutionarily distant organisms: S. cerevisiae, C. elegans, E. coli, A. thaliana, D. melanogaster, and H. sapiens. We use genomic sequence information to connect these data and compare global and modular properties of the transcription programs. Linking genes whose expression profiles are similar, we find that for all organisms the connectivity distribution follows a power-law, highly connected genes tend to be essential and conserved, and the expression program is highly modular. We reveal the modular structure by decomposing each set of expression data into coexpressed modules. Functionally related sets of genes are frequently coexpressed in multiple organisms. Yet their relative importance to the transcription program and their regulatory relationships vary among organisms. Our results demonstrate the potential of combining sequence and expression data for improving functional gene annotation and expanding our understanding of how gene expression and diversity evolved. PMID- 14737188 TI - Science and technology communication for development. PMID- 14737189 TI - The proteasome and the delicate balance between destruction and rescue. PMID- 14737190 TI - Protein interaction networks by proteome peptide scanning. AB - A substantial proportion of protein interactions relies on small domains binding to short peptides in the partner proteins. Many of these interactions are relatively low affinity and transient, and they impact on signal transduction. However, neither the number of potential interactions mediated by each domain nor the degree of promiscuity at a whole proteome level has been investigated. We have used a combination of phage display and SPOT synthesis to discover all the peptides in the yeast proteome that have the potential to bind to eight SH3 domains. We first identified the peptides that match a relaxed consensus, as deduced from peptides selected by phage display experiments. Next, we synthesized all the matching peptides at high density on a cellulose membrane, and we probed them directly with the SH3 domains. The domains that we have studied were grouped by this approach into five classes with partially overlapping specificity. Within the classes, however, the domains display a high promiscuity and bind to a large number of common targets with comparable affinity. We estimate that the yeast proteome contains as few as six peptides that bind to the Abp1 SH3 domain with a dissociation constant lower than 100 microM, while it contains as many as 50-80 peptides with corresponding affinity for the SH3 domain of Yfr024c. All the targets of the Abp1 SH3 domain, identified by this approach, bind to the native protein in vivo, as shown by coimmunoprecipitation experiments. Finally, we demonstrate that this strategy can be extended to the analysis of the entire human proteome. We have developed an approach, named WISE (whole interactome scanning experiment), that permits rapid and reliable identification of the partners of any peptide recognition module by peptide scanning of a proteome. Since the SPOT synthesis approach is semiquantitative and provides an approximation of the dissociation constants of the several thousands of interactions that are simultaneously analyzed in an array format, the likelihood of each interaction occurring in any given physiological settings can be evaluated. WISE can be easily extended to a variety of protein interaction domains, including those binding to modified peptides, thereby offering a powerful proteomic tool to help completing a full description of the cell interactome. PMID- 14737185 TI - Loss of olfactory receptor genes coincides with the acquisition of full trichromatic vision in primates. AB - Olfactory receptor (OR) genes constitute the molecular basis for the sense of smell and are encoded by the largest gene family in mammalian genomes. Previous studies suggested that the proportion of pseudogenes in the OR gene family is significantly larger in humans than in other apes and significantly larger in apes than in the mouse. To investigate the process of degeneration of the olfactory repertoire in primates, we estimated the proportion of OR pseudogenes in 19 primate species by surveying randomly chosen subsets of 100 OR genes from each species. We find that apes, Old World monkeys and one New World monkey, the howler monkey, have a significantly higher proportion of OR pseudogenes than do other New World monkeys or the lemur (a prosimian). Strikingly, the howler monkey is also the only New World monkey to possess full trichromatic vision, along with Old World monkeys and apes. Our findings suggest that the deterioration of the olfactory repertoire occurred concomitant with the acquisition of full trichromatic color vision in primates. PMID- 14737191 TI - Multiple apoptotic caspase cascades are required in nonapoptotic roles for Drosophila spermatid individualization. AB - Spermatozoa are generated and mature within a germline syncytium. Differentiation of haploid syncytial spermatids into single motile sperm requires the encapsulation of each spermatid by an independent plasma membrane and the elimination of most sperm cytoplasm, a process known as individualization. Apoptosis is mediated by caspase family proteases. Many apoptotic cell deaths in Drosophila utilize the REAPER/HID/GRIM family proapoptotic proteins. These proteins promote cell death, at least in part, by disrupting interactions between the caspase inhibitor DIAP1 and the apical caspase DRONC, which is continually activated in many viable cells through interactions with ARK, the Drosophila homolog of the mammalian death-activating adaptor APAF-1. This leads to unrestrained activity of DRONC and other DIAP1-inhibitable caspases activated by DRONC. Here we demonstrate that ARK- and HID-dependent activation of DRONC occurs at sites of spermatid individualization and that all three proteins are required for this process. dFADD, the Drosophila homolog of mammalian FADD, an adaptor that mediates recruitment of apical caspases to ligand-bound death receptors, and its target caspase DREDD are also required. A third apoptotic caspase, DRICE, is activated throughout the length of individualizing spermatids in a process that requires the product of the driceless locus, which also participates in individualization. Our results demonstrate that multiple caspases and caspase regulators, likely acting at distinct points in time and space, are required for spermatid individualization, a nonapoptotic process. PMID- 14737195 TI - Channelling evolution: canalization and the nervous system. PMID- 14737196 TI - Role of Saccharomyces single-stranded DNA-binding protein RPA in the strand invasion step of double-strand break repair. AB - The single-stranded DNA (ssDNA)-binding protein replication protein A (RPA) is essential for both DNA replication and recombination. Chromatin immunoprecipitation techniques were used to visualize the kinetics and extent of RPA binding following induction of a double-strand break (DSB) and during its repair by homologous recombination in yeast. RPA assembles at the HO endonuclease cut MAT locus simultaneously with the appearance of the DSB, and binding spreads away from the DSB as 5' to 3' exonuclease activity creates more ssDNA. RPA binding precedes binding of the Rad51 recombination protein. The extent of RPA binding is greater when Rad51 is absent, supporting the idea that Rad51 displaces RPA from ssDNA. RPA plays an important role during RAD51-mediated strand invasion of the MAT ssDNA into the donor sequence HML. The replication-proficient but recombination-defective rfa1-t11 (K45E) mutation in the large subunit of RPA is normal in facilitating Rad51 filament formation on ssDNA, but is unable to achieve synapsis between MAT and HML. Thus, RPA appears to play a role in strand invasion as well as in facilitating Rad51 binding to ssDNA, possibly by stabilizing the displaced ssDNA. PMID- 14737199 TI - PLoS Biology in action. PMID- 14737198 TI - Long-lasting novelty-induced neuronal reverberation during slow-wave sleep in multiple forebrain areas. AB - The discovery of experience-dependent brain reactivation during both slow-wave (SW) and rapid eye-movement (REM) sleep led to the notion that the consolidation of recently acquired memory traces requires neural replay during sleep. To date, however, several observations continue to undermine this hypothesis. To address some of these objections, we investigated the effects of a transient novel experience on the long-term evolution of ongoing neuronal activity in the rat forebrain. We observed that spatiotemporal patterns of neuronal ensemble activity originally produced by the tactile exploration of novel objects recurred for up to 48 h in the cerebral cortex, hippocampus, putamen, and thalamus. This novelty induced recurrence was characterized by low but significant correlations values. Nearly identical results were found for neuronal activity sampled when animals were moving between objects without touching them. In contrast, negligible recurrence was observed for neuronal patterns obtained when animals explored a familiar environment. While the reverberation of past patterns of neuronal activity was strongest during SW sleep, waking was correlated with a decrease of neuronal reverberation. REM sleep showed more variable results across animals. In contrast with data from hippocampal place cells, we found no evidence of time compression or expansion of neuronal reverberation in any of the sampled forebrain areas. Our results indicate that persistent experience-dependent neuronal reverberation is a general property of multiple forebrain structures. It does not consist of an exact replay of previous activity, but instead it defines a mild and consistent bias towards salient neural ensemble firing patterns. These results are compatible with a slow and progressive process of memory consolidation, reflecting novelty-related neuronal ensemble relationships that seem to be context- rather than stimulus-specific. Based on our current and previous results, we propose that the two major phases of sleep play distinct and complementary roles in memory consolidation: pretranscriptional recall during SW sleep and transcriptional storage during REM sleep. PMID- 14737201 TI - RNAi therapeutics: how likely, how soon? PMID- 14737208 TI - Intrarater and Interrater Reliability of the Beighton and Horan Joint Mobility Index. AB - OBJECTIVE: Clinicians may benefit from using a joint mobility index to screen for individuals on the high end of the spectrum of joint laxity (ie, those with generalized joint laxity), which may be associated with musculoskeletal complaints. Reliability of the Beighton and Horan Joint Mobility Index (BHJMI) has not been reported in the literature. Our purpose was to determine intrarater and interrater reliability of (1) composite BHJMI scores (the overall score from 0 to 9), and (2) categorized scores, the BHJMI scores in 3 categories (0 to 2, 3 to 4, and 5 to 9) DESIGN AND SETTING: This was an intrarater and interrater reliability study. Data were collected in an academic physical therapy department and in a high school. SUBJECTS: Forty-two (intrarater) and 36 (interrater) female volunteers, aged 15 to 45 years. MEASUREMENTS: Subjects were screened using the BHJMI. Percentage agreement and the Spearman rho were used to analyze BHJMI composite and category scores. RESULTS: The percentage agreement and the Spearman rho for intrarater and interrater reliability of BHJMI composite scores were 69% and.86 and 51% and.87, respectively. The percentage agreement and the Spearman rho for intrarater and interrater reliability of the category scores were 81% and.81 and 89% and.75, respectively. CONCLUSIONS: Reliability of the BHJMI was good to excellent in screening for generalized joint laxity in females aged 15 to 45 years. PMID- 14737209 TI - Physiologic Screening Test for Eating Disorders/Disordered Eating Among Female Collegiate Athletes. AB - OBJECTIVE: To develop and evaluate a physiologic screening test specifically designed for collegiate female athletes engaged in athletic competition or highly athletic performances in order to detect eating disorders/disordered eating. No such physiologically based test currently exists. METHODS: Subjects included 148 (84.5%) of 175 volunteer, National Collegiate Athletic Association Division I (n = 92), club (n = 15), and dance team (n = 41) athletes 18 to 25 years old who attended a large, Midwestern university. Participants completed 4 tests: 2 normed for the general population (Eating Disorders Inventory-2 and Bulimia Test Revised); a new physiologic test, developed and pilot tested by the investigators, called the Physiologic Screening Test; and the Eating Disorder Exam 12.0D, a structured, validated, diagnostic interview used for criterion validity. RESULTS: The 18-item Physiologic Screening Test produced the highest sensitivity (87%) and specificity (78%) and was superior to the Eating Disorders Inventory-2 (sensitivity = 62%, specificity = 74%) and Bulimia Test-Revised (sensitivity = 27%, specificity = 99%). A substantial number (n = 51, 35%) of athletes were classified as eating disordered/disordered eating. CONCLUSIONS: The Physiologic Screening Test should be considered for screening athletes for eating disorders/disordered eating. The Physiologic Screening Test seems to be a viable alternative to existing tests because it is specifically designed for female athletes, it is brief (4 measurements and 14 items), and validity is enhanced and response bias is lessened because the purpose is less obvious, especially when included as part of a mandatory preparticipation examination. PMID- 14737210 TI - Association of Menstrual-Cycle Hormone Changes with Anterior Cruciate Ligament Laxity Measurements. AB - OBJECTIVE: To determine whether anterior cruciate ligament (ACL) laxity (as evaluated with the KT-2000 and radiographic measures) is associated with concentrations of reproductive hormones during the menstrual cycle and whether the KT-2000 knee arthrometer is a valid measurement technique, compared with radiographic techniques. DESIGN AND SETTING: A within-subjects linear model was used. Venipuncture was conducted in an exercise science laboratory. The KT-2000 and radiographic measurements were performed in a hospital radiology laboratory. SUBJECTS: Twelve females presented with a dominant right leg free of injury. They were mild to moderately active and had a 12-month history of normal menstrual cycles (28-35 days). Subjects had not used hormonal therapy for the previous 3 months. MEASUREMENTS: Subjects were tested at the onset of menses, near ovulation, and on day 23 of the midluteal phase of the menstrual cycle. At each session, 14 mL of blood was obtained by venipuncture. Blood samples were analyzed via radioimmunoassay to determine the relative levels of each hormone. Anterior cruciate ligament laxity was simultaneously measured by KT-2000 and radiographic techniques. RESULTS: Anterior cruciate ligament laxity measurements were significantly greater with the KT-2000 than with radiographic measurement. No significant differences were noted between menstrual-cycle phase and ACL laxity for either method. The concentration of luteinizing hormone was negatively correlated with laxity at the onset of menses using the radiographic technique. CONCLUSIONS: We found no associations between follicular-, ovulatory-, and luteal phase hormonal concentrations and ACL laxity as measured on the KT-2000 and radiographs; no effects of menstrual-cycle phase on ACL laxity as measured by the KT-2000 and radiographs; and significant differences between KT-2000 and radiographic measures of ACL laxity. PMID- 14737211 TI - Reliability of Joint Position Sense and Force-Reproduction Measures During Internal and External Rotation of the Shoulder. AB - OBJECTIVE: To determine the reliability of 2 common measures of proprioception. DESIGN AND SETTING: Joint position sense (JPS) and force reproduction (FR) were measured in the dominant shoulder using internal-rotation (IR) and external rotation (ER) target angles on 2 consecutive days. SUBJECTS: Thirty-one healthy subjects (age = 22.0 +/- 2.8 years, height = 171.8 +/- 9.2 cm, mass = 69.5 +/- 15.9 kg) who did not regularly compete in overhand sports volunteered to participate in the study. MEASUREMENTS: Error scores were calculated at 2 target angles by averaging the absolute difference of 3 trials of JPS and FR. Reliability was determined by comparing the error scores obtained on 2 consecutive days. RESULTS: The inclinometer was found to be a reliable instrument as both intertester (.999) and intratester (.999) intraclass correlation coefficients were high. The JPS and FR measurements were also found to be reliable, with intraclass correlation coefficients ranging from.978 to.984. No differences were observed between trials for either measure. CONCLUSIONS: The inclinometer was a reliable instrument and can provide an affordable and accurate measure of range of motion and JPS. Both JPS and FR were also reliable measures of proprioception in the shoulder. Further research is needed to identify the specific mechanism of proprioception during these tasks. PMID- 14737212 TI - Ground Reaction Forces Among Gymnasts and Recreational Athletes in Drop Landings. AB - OBJECTIVE: To compare vertical ground reaction forces among gymnasts and recreational athletes during drop landings from 30-, 60-, and 90-cm heights. DESIGN AND SETTING: Two subject groups, intercollegiate gymnasts and college-aged recreational athletes, participated in this study. Subjects completed 10 landing trials onto a force platform at each height. SUBJECTS: Ten female competitive gymnasts (height = 1.57 +/- 0.02 m, mass = 55.4 +/- 7.3 kg) and 10 female recreational athletes (height = 1.63 +/- 0.06 m, mass = 59.6 +/- 4.9 kg) volunteered for this study. MEASUREMENTS: Measurements of first peak-force magnitude (F1), time to F1 (T1), impulse to F1, second peak-force magnitude (F2), time to F2 (T2), and impulse to F2 were compiled to describe the ground reaction force profile for each trial at 30-, 60-, and 90-cm platform heights. A 2 x 3 (group x height) mixed-factors analysis of variance was calculated for each of the 6 variables. RESULTS: The group-by-height interaction was significant for F1, F2, and impulse to F2. Tukey post hoc analyses revealed significantly higher values for the gymnasts than for the recreational athletes at 60- and 90-cm heights for F1 and F2 magnitudes. Differences between groups for T1, T2, impulse to F1, and impulse to F2 were not statistically significant at any height. CONCLUSIONS: Drop landings performed by female gymnasts at 60- and 90-cm heights exhibited higher vertical ground reaction forces than drop landings performed by female recreational athletes. High ground reaction forces experienced by gymnasts during landings may contribute to the incidence of lower extremity injuries. PMID- 14737213 TI - Comparison of 3 Methods to Assess Urine Specific Gravity in Collegiate Wrestlers. AB - OBJECTIVE: To investigate the reliability and validity of refractometry, hydrometry, and reagent strips in assessing urine specific gravity in collegiate wrestlers. DESIGN AND SETTING: We assessed the reliability of refractometry, hydrometry, and reagent strips between 2 trials and among 4 testers. The validity of hydrometry and reagent strips was assessed by comparison with refractometry, the criterion measure for urine specific gravity. SUBJECTS: Twenty-one National Collegiate Athletic Association Division III collegiate wrestlers provided fresh urine samples. MEASUREMENTS: Four testers measured the specific gravity of each urine sample 6 times: twice by refractometry, twice by hydrometry, and twice by reagent strips. RESULTS: Refractometer measurements were consistent between trials (R =.998) and among testers; hydrometer measurements were consistent between trials (R =.987) but not among testers; and reagent-strip measurements were not consistent between trials or among testers. Hydrometer (1.018 +/- 0.006) and reagent-strip (1.017 +/- 0.007) measurements were significantly higher than refractometer (1.015 +/- 0.006) measurements. Intraclass correlation coefficients were moderate between refractometry and hydrometry (R =.869) and low between refractometry and reagent strips (R =.573). The hydrometer produced 28% false positives and 2% false negatives, and reagent strips produced 15% false positives and 9% false negatives. CONCLUSIONS: Only the refractometer should be used to determine urine specific gravity in collegiate wrestlers during the weight certification process. PMID- 14737214 TI - Volume Decreases After Elevation and Intermittent Compression of Postacute Ankle Sprains Are Negated by Gravity-Dependent Positioning. AB - OBJECTIVE: Elevation and intermittent compression are routinely prescribed after soft tissue injury. Individuals must, however, resume activity in an upright position. The effect of gravity-dependent positioning after elevation and intermittent compression has not been extensively examined. Our purpose was to examine the effects of gravity-dependent positioning after elevation and intermittent compression on the volume of injured ankles. DESIGN AND SETTING: Ankle-injured subjects were randomly assigned to 1 of 2 treatment groups: elevation or elevation and intermittent compression. Each treatment lasted 30 minutes. SUBJECTS: Twelve college students with inversion ankle sprains 2 to 4 days earlier participated. MEASUREMENTS: Measurements of ankle volume were performed before treatment and at 5, 10, 15, 20, 25, 30, 45, and 60 minutes after treatment. RESULTS: Regardless of treatment, ankle volume decreased (by 17.25 +/- 4.05 mL) between the pretreatment measurement and the immediate posttreatment measurement (P <.05). The effects of both treatments, however, lasted less than 5 minutes after the limb was returned to a gravity-dependent position. CONCLUSIONS: These results suggest that elevation or elevation and intermittent compression do not effectively decrease ankle volume for a prolonged period of time in patients with postacute ankle sprains. PMID- 14737215 TI - Effects of Cool-Water Immersion and High-Voltage Electric Stimulation for 3 Continuous Hours on Acute Edema in Rats. AB - OBJECTIVE: Cool-water immersion (CWI) at 12.8 degrees C (55 degrees F), cathodal high-voltage pulsed current (CHVPC) at 120 pulses per second and 90% of visible motor threshold, or the combination of CWI and CHVPC, applied 30 minutes on, 30 minutes off for 4 hours, are known to curb edema formation after blunt trauma to the hind limbs of rats. Our purpose was to examine the effects of extending treatment times to 3 continuous hours after blunt trauma to the hind limbs of rats. DESIGN AND SETTING: A randomized, parallel-groups design of 22 subjects was used. Volumes of traumatized limbs, randomly assigned to CWI (n = 7), CHVPC (n = 8), or CWI followed by CHVPC (n = 7) were compared with those of injured but untreated limbs with analysis of variance. SUBJECTS: Twenty-two anesthetized Zucker lean rats (mass = 293 +/- 27 g). MEASUREMENTS: We measured limb volumes immediately before and after trauma and every 30 minutes over the 4-hour experiment. RESULTS: Volumes of treated limbs of all 3 experimental groups were smaller than those of untreated limbs (P <.05). No treatment was more effective than another. CONCLUSIONS: Exposure to either 3 hours of CWI or CHVPC or to 1 hour of CWI followed by 2 hours of CHVPC effectively curbed edema after blunt injury. These results suggest that these common treatments are effective only during application and hint that application should be maintained throughout the period during which edema is forming. PMID- 14737216 TI - Efficacy of Rugby Headgear in Attenuating Repetitive Linear Impact Forces. AB - OBJECTIVE: To determine the effectiveness of rugby headgear and the effect of impact site and headgear type on attenuating repetitive linear impact forces. DESIGN AND SETTING: We obtained 10 headgear of 2 different types (n = 20) from 2 popular manufacturers: type I was Honeycomb headgear and type II was Vanguard headgear. Both headgear types were approved by the International Rugby Board. Headgear were tested according to National Operating Committee on Standards for Athletic Equipment protocols with one modification. The headgear were dropped from a height of 30 cm according to the International Rugby Board standards. Headgear were tested at a National Athletic Equipment Reconditioners Association facility. MEASUREMENTS: We recorded peak acceleration of gravity (g) and Gadd Severity Index readings for each impact. The design of the study was 4 one-way, within-subjects, repeated-measures analyses of variance with alpha =.05. Descriptive statistics were calculated for all categoric variables. RESULTS: Peak g and Gadd Severity Index increased with repetitive impacts, resulting in a headgear's decreased ability to attenuate linear impact forces. Attenuation differed significantly between headgear type I and type II at both impact sites and between the parietal-lateral and occipital impact sites for both headgear types. CONCLUSIONS: The headgear initially attenuated impact forces from a drop height of 30 cm. However, over 10 drops, both headgear decreased in their ability to attenuate repetitive linear impact forces. Decreased attenuation in the occipital region as well as significant differences in attenuation abilities between headgear types support the need for further investigation to examine the efficacy of rugby headgear. PMID- 14737217 TI - Program Directors' and Clinical Instructors' Perceptions of Important Clinical Instructor Behavior Categories in the Delivery of Athletic Training Clinical Instruction. AB - OBJECTIVE: To investigate the perceived importance of clinical instructors' behaviors and behavior categories in the roles of program director (PD) and clinical instructor (CI) and to ascertain the relative importance of these items within each role. DESIGN AND SETTING: From the literature, we developed a questionnaire, validated by a panel of experts, to collect data regarding the perceived importance of 30 specific CI behavior statements within 5 categories (instructional, interpersonal, evaluative, professional, and personal). The instrument used in the study had a Cronbach alpha of.92. SUBJECTS: Independent groups of 75 PDs and 242 CIs from Commission on Accreditation of Allied Health Education Programs-accredited entry-level undergraduate athletic training programs returned usable surveys. MEASUREMENTS: We computed mean and variation measures for each behavior and category and analyzed these items for between-role and within-role category differences. RESULTS: Program directors and CIs differed significantly in the perceived importance of the evaluative category of CI behaviors as well as in 3 specific evaluative behaviors. Program directors and CIs did not differ in the other 4 categories. For within-role groupings, we noted significant differences of perceived importance among the behavioral categories. CONCLUSIONS: Program directors and CIs perceived all 5 categories to be very important, and they should work to demonstrate these behaviors in clinical education settings. Collaboration between the groups enhances the understanding of role responsibility in the delivery of athletic training clinical instruction. PMID- 14737218 TI - Educational Reform in Athletic Training: A Policy Analysis. AB - OBJECTIVE: To apply a policy-analysis framework to the athletic training educational reform policy that will be fully implemented by January 2004. DATA SOURCES: Policy analysis is not a specific science. No one framework exists for conducting all policy analyses. I used literature from the education, policy analysis, and athletic training fields as data sources to provide background and to create a framework from which to conduct the policy analysis. DATA SYNTHESIS: Once the policy-analysis framework was selected, I began data synthesis, using several athletic training sources in support of the findings. The tension among the myriad stakeholders in this policy is clear. Although many see the benefits of accreditation, some experience hardships from the imposed policy. CONCLUSIONS/RECOMMENDATIONS: Of the 4 possible alternatives suggested, following the route currently under implementation (Committee on Accreditation of Health Education Programs accreditation) was the most agreeable solution. The goals as stated by the policy makers are attained by the policy. However, issues within the accreditation process itself need to be addressed. Of the many stakeholders in the reform effort, some will see little gain and have many hardships imposed on them. As the policy is implemented, unintended implications will likely arise, as with any new policy. Thus, I recommend that the National Athletic Trainers' Association develop a system dedicated solely to reducing the hardships faced by many of its members as the policy is implemented. PMID- 14737219 TI - Gene expression signature of fibroblast serum response predicts human cancer progression: similarities between tumors and wounds. AB - Cancer invasion and metastasis have been likened to wound healing gone awry. Despite parallels in cellular behavior between cancer progression and wound healing, the molecular relationships between these two processes and their prognostic implications are unclear. In this study, based on gene expression profiles of fibroblasts from ten anatomic sites, we identify a stereotyped gene expression program in response to serum exposure that appears to reflect the multifaceted role of fibroblasts in wound healing. The genes comprising this fibroblast common serum response are coordinately regulated in many human tumors, allowing us to identify tumors with gene expression signatures suggestive of active wounds. Genes induced in the fibroblast serum-response program are expressed in tumors by the tumor cells themselves, by tumor-associated fibroblasts, or both. The molecular features that define this wound-like phenotype are evident at an early clinical stage, persist during treatment, and predict increased risk of metastasis and death in breast, lung, and gastric carcinomas. Thus, the transcriptional signature of the response of fibroblasts to serum provides a possible link between cancer progression and wound healing, as well as a powerful predictor of the clinical course in several common carcinomas. PMID- 14737222 TI - A spectroelectrochemical study on perylene cation radical in polymer microchannel microelectrode chips. AB - Polymer microchannel chips (depth 20 microm and width 100 microm) integrated with band electrodes were fabricated by photolithography and imprinting methods, and applied to a spectroelectrochemical study on the cation radical of perylene (Pe). A propylene carbonate solution of Pe was brought into the channel chip by pressure driven flow and Pe was oxidized at the working band electrode (WE) in the channel. Simultaneously, absorption measurements of the solution phase in the downstream side of the electrode (30 microm from WE) were conducted on the basis of space resolved spectroscopy. The decrease in the absorbance of Pe at 438 nm upon electrolysis accompanied an appearance of the absorption band around 538 nm, which was assigned to that of the Pe cation radical. When the perylene solution was introduced to the microchip at a slow flow velocity, the dimer cation radical of Pe was shown to be produced in the channel chip. The formation and disappearance processes of the monomer and dimer cation radicals of Pe in the channel were followed by flow velocity and position dependencies of the absorption spectra. PMID- 14737221 TI - Force and Compliance Measurements on Living Cells Using Atomic Force Microscopy (AFM). AB - We describe the use of atomic force microscopy (AFM) in studies of cell adhesion and cell compliance. Our studies use the interaction between leukocyte function associated antigen-1 (LFA-1)/intercellular adhesion molecule-1 (ICAM-1) as a model system. The forces required to unbind a single LFA-1/ICAM-1 bond were measured at different loading rates. This data was used to determine the dynamic strength of the LFA-1/ICAM-1 complex and characterize the activation potential that this complex overcomes during its breakage. Force measurements acquired at the multiple- bond level provided insight about the mechanism of cell adhesion. In addition, the AFM was used as a microindenter to determine the mechanical properties of cells. The applications of these methods are described using data from a previous study. PMID- 14737224 TI - Overoxidation of carbon-fiber microelectrodes enhances dopamine adsorption and increases sensitivity. AB - The voltammetric responses of carbon-fiber microelectrodes with a 1.0 V and a 1.4 V anodic limit were compared in this study. Fast-scan cyclic voltammetry was used to characterize the response to dopamine and several other neurochemicals. An increase in the adsorption properties of the carbon fiber leads to an increase in sensitivity of 9 fold in vivo. However the temporal response of the sensor is slower with the more positive anodic limit. Increased electron transfer kinetics also causes a decrease in the relative sensitivity for dopamine vs. other neurochemicals, and a change in their cyclic voltammograms. Stimulated release in the caudate-putamen was pharmacologically characterized in vivo using Ro-04-1284 and pargyline, and was consistent with that expected for dopamine. PMID- 14737223 TI - Development of a dual luciferase reporter screening assay for the detection of synthetic glucocorticoids in animal tissues. AB - Synthetic glucocorticoids belong to the most frequently administered drugs in livestock production. These synthetic hormones are employed for therapeutic purposes against inflammatory reactions, disorders of the musculoskeletal system, bovine ketosis and many other diseases of farm animals. A widespread illegal use of synthetic glucocorticoids to improve feed intake and weight gain has also been observed. To enforce the residue limits imposed on glucocorticoid drugs and preclude their illicit administration as growth promoters, it is necessary to establish high throughput analytical methods that can be applied to the screening of animal tissues. Here, we developed a dual luciferase reporter assay that detects residues or contaminants with glucocorticoid activity. This screening assay is performed by transfection of human cell lines with two reporter constructs followed by the measurement of two distinct luminescence signals, one of which serves as internal control to correct for assay variabilities and unspecific matrix effects. The limit of detection (1.25 microg for dexamethasone in liver) depends on the biological potency of each synthetic glucocorticoid but, with all drugs tested, the maximal response reaches a 20 to 30 fold induction of luciferase activity. In combination with an appropriate sample clean-up method (recovery of 82%), this luciferase assay has been applied to the analysis of liver samples from calves treated with a single therapeutic injection of either dexamethasone or flumethasone. Thus, the dual luciferase reporter assay provides a new screening tool to detect unwanted glucocorticoid activities in animal tissues or other crude biological samples without knowledge of the precise chemical entity of the parent compounds or their metabolites. PMID- 14737225 TI - Nitrite reduction and detection at a carbon paste electrode containing hemoglobin and colloidal gold. AB - A novel renewable reagentless nitrite biosensor based on the direct electron transfer of hemoglobin (Hb) and a new sensing mechanism was proposed by combining the advantageous features of colloidal gold nanoparticle and carbon paste technology. The direct electrochemistry of immobilized Hb displayed a pair of redox peaks with a formal potential of -42 mV (vs. NHE) in 0.2 mol dm(-3) NaAc HAc buffer (pH 5.5). The immobilized Hb displayed an excellent response to the reduction of NO2(-) with one interfacial charge transfer followed by a chemical reaction (EC) mechanism. Under optimal conditions, the interfacial EC process could be used for the sensitive determination of NO2(-) with a linear range from 0.1 to 9.7 micromol dm(-3) and a detection limit of 0.06 [micro sign]mol dm(-3) at 3sigma. The amperometric determination of high concentrations of NO2(-) based on the irreversible reduction of NO could be performed at pH 4.0 with a linear range from 0.1 to 1.2 mmol dm(-3). The surface of biosensor could be renewed quickly and reproducibly by a simple polish step. The biosensor has been used satisfactorily for nitrite determination in native water samples. PMID- 14737226 TI - Moment analysis of retention equilibrium, mass transfer kinetics, and thermodynamic properties in reversed-phase liquid chromatography using phenyl bonded silica gel. AB - Information about retention equilibrium, mass transfer kinetics, and related thermodynamic properties of chromatography using phenyldimethylsilyl (Ph)-silica gel was derived by moment analysis of pulse response peak profiles. The results for the Ph-silica gel were compared with those for octadecyldimethylsilyl (C18) silica gel. Some parameters characterizing the chromatographic behavior of the two stationary phases were correlated with the hydrophobic surface area of sample molecules. Surface diffusion had a predominant role for intraparticle diffusion. An enthalpy-entropy compensation was established for both the retention equilibrium and surface diffusion. A linear correlation was observed between the logarithm of surface diffusion coefficient (Ds) and that of retention equilibrium constant (Ka), suggesting the establishment of a linear free energy relationship. The ratio of Ds to molecular diffusivity (Dm) decreased with increasing Ka and was correlated by a single curved line. The value of Ds was of the same order of magnitude with Dm when Ka became negligible. These results suggest the presence of a sort of correlation between surface diffusion and molecular diffusion and the restriction of the molecular mobility by surface diffusion due to the retention strength. PMID- 14737227 TI - Capillary array electrophoresis with confocal fluorescence rotary scanner. AB - A capillary array electrophoresis system with a rotary confocal fluorescence scanner is reported. A high speed direct current rotary motor, combined with a rotary encoder and a reflection mirror, has been designed to direct the excitation laser beam precisely to a round array of capillaries which are symmetrically distributed around the motor. The rotary encoder is introduced to accurately orientate the position of each capillary and its output signal triggers the data acquisition system to record the fluorescence signal corresponding to each capillary. Separation of enantiomers of glutamic acid, methionine and tryptophan with different additives are demonstrated by this system. The experimental results indicate that this setup can be used to optimize separation methods for capillary electrophoresis as quickly as possible. PMID- 14737228 TI - Trace ion analysis of sea water by capillary electrophoresis: determination of strontium and lithium pre-concentrated by transient isotachophoresis. AB - The applicability of capillary zone electrophoresis (CZE) to ions having relatively low natural occurrences in sea water is limited by method's relatively poor concentration detection sensitivity. A combination of CZE with indirect UV detection and transient isotachophoresis (tITP) pre-concentration was developed to evolve the CZE practical utility towards the quantitative determination of the minor sea water cationic components, strontium and lithium. The ITP stacking criterion at the initial stage of a CZE separation was met by taking a highly mobile sodium, the principle matrix cation, to perform the role of a leading ion, whereas the moderately mobile sample macrocomponents, Ca2+ and Mg2+, acted as the terminating ion. The carrier electrolyte, consisting of 10 mM 4-methylbenzylamine and 1.5 mM citric acid at pH 4.8, was found to be optimal to accommodate both analyte cations in the ITP range and then separate them in the CZE mode, with relative standard deviations for migration times from 0.06-0.15% and for peak areas from 4-8%. The limits of detection were 1.3 mg l(-1) Sr2+ and 0.12 mg l(-1) Li+. The developed method was applied to the analysis of a surface sea water sample and a sea water reference material. The results were in good agreement with those obtained by inductively coupled plasma atomic emission spectroscopy (ICP-AES) and electrothermal atomic absorption spectrometry (ET-AAS). PMID- 14737229 TI - Analysis of benzene ethylamine derivatives in urine using the programmable dynamic liquid-phase microextraction (LPME) device. AB - An automated LPME device for a dynamic LPME method was manufactured and its extraction efficiency was tested using spiked urine samples. The developed home made LPME device was a programmable automated syringe dispenser to overcome deteriorating precision and difficulties in manually manipulating the plunger repeatedly. To establish the optimum parameters for benzene ethylamines, the effects of sampling volume, solvent volume, pH, salt-effect, choice of solvents, plunger speed, and number of samplings were investigated. Good repeatabilities for the extraction of mephentermine, ephedrine, methoxyphenamine, selegiline, and bupropion were obtained and the RSD values were 2.4, 1.9, 1.3, 1.6 and 1.5% at a concentration of 3 microg mL(-1) in spiked urine samples, respectively. The limit of detection was below 0.05 microg mL(-1) for the investigated drugs. This developed device for LPME analysis gave good validation results and improved convenience. PMID- 14737230 TI - Determination of unreacted 2,4-toluene diisocyanate (2,4TDI) and 2,6-toluene diisocyanate (2,6TDI) in foams at ultratrace level by using HPLC-CIS-MS-MS. AB - Isocyanates can cause occupational asthma. By using available HPLC-UVF methods, isocyanates can be quantified only at levels above 1% of the Permissible Exposure Limits (PEL). Once sensitized, workers can react to concentrations below these limits of detection (LOD) making these methods insufficiently sensitive to adequately evaluate trace amounts of isocyanates present in air or in materials at safe levels for sensitized workers. This article describes a novel method for isocyanate analysis allowing the quantification of 2,4TDI and 2,6TDI monomers at very low concentrations using HPLC-CIS-MS-MS. The method's sensitivity increases with a decrease in the alkali radius. The LOD is 0.039 ng mL(-1) for 2,4TDI and 0.100 ng mL(-1) for 2,6TDI in solution when lithium is the alkali adduct, which is 20 times more sensitive than HPLC-UVF method. This new method allows determination in foam at levels of 0.078 ng g(-1) for 2,4TDI and 0.200 ng g(-1) for 2,6TDI respectively, for a 0.5 g foam sample. This is more than 100 times more sensitive than other methods for determining free monomers in solid materials. Analytical reproducibility and precision are better than 92% and 93% for both diisocyanate monomers. The use of HPLC-UVF conventional method failed to detect unreacted isocyanates in foam samples, but TDI monomers were quantified by HPLC-CIS-MS-MS. PMID- 14737231 TI - Matrix-assisted laser desorption/ionization mass spectrometry with re-engineered 2,5-dihydroxybenzoic acid derivative. AB - Dihydroxybenzoic acid was modified to three analogues (M2, M4 and M6). The analogues exhibited specific properties that resulted in enhancement of analyte signal intensity with or without addition of iodine compared to the underivatized parent. Addition of iodine to M2, an ester of dihydroxybenzoic acid that had a terminal double bond in the alkyl chain, resulted in peak intensities comparable to the parent, indicating that iodine interaction across the double bond resulted in enhancement although the exact mechanism is not fully understood. No enhancement on addition of iodine was observed for M4, which had a long alkyl chain that contained no double bonds. The alkyl chain allowed micelle formation in solution, which in turn allowed more uniform analyte-to-matrix mixing. The final analogue combined the long alkyl chain of M4 with the double bond of M2 and exhibited either similar peak intensities to that of dihydroxybenzoic acid or better. Micelle formation in solution was examined using spectroscopy and in the solid by reflective microscopy. The standard deviation from spot to spot was considerably lower relative to dihydroxybenzoic acid (RSD 3.4%vs. 14.2%). Unlike dihydroxybenzoic acid, the novel matrix M6 was able to yield characteristic peaks for analytes such as ubiquitin. PMID- 14737232 TI - An investigation of the chemical stability of arsenosugars in basic environments using IC-ICP-MS and IC-ESI-MS/MS. AB - This paper evaluates the chemical stability of four arsenosugars using tetramethylammonium hydroxide (TMAOH) as an extraction solvent. This solvent was chosen because of the near quantitative removal of these arsenicals from difficult to extract seafood (oysters and shellfish). Four arsenosugars (3-[5' deoxy-5'-(dimethylarsinoyl)-beta-ribofuranosyloxy]-2-hydroxypropylene glycol- As(328), 3-5'-deoxy-5'-(dimethylarsinoyl)-beta-ribofuranosyloxy]-2 hydroxypropanesulfonic acid--As(392), 3-[5'-deoxy-5'-(dimethylarsinoyl)-beta ribofuranosyloxy]-2-hydroxypropyl hydrogen sulfate--As(408), and 3-[5'-deoxy-5' (dimethylarsinoyl)-beta-ribofuranosyloxy]-2-hydroxypropyl-2,3-hydroxypropyl phosphate--As(482)) were evaluated. The stability of these four arsenosugars were studied independently in a solution of 2.5% TMAOH at 60 degrees C over a period of up to 8 h. Two arsenosugars, As(328) and As(392), were found to be relatively stable in this solution for up to 8 h. However, As(408) and As(482) formed detectable quantities of dimethylarsinic acid (DMAA) and As(328) within 0.5 and 2 h, respectively. It was found that 97% of As(408) degrades after 8 h of treatment producing 3.4 times as much DMAA as As(328). This is contrary to As(482), which produces 13 times as much As(328) as DMAA and only 37% of the As(482) was converted by the 8 h treatment at 60 degrees C. These degradation products led to the investigation of weaker TMAOH extraction solvents. Three different concentrations (2.5%, 0.83% and 0.25%) were used to determine the effect of TMAOH concentration on the degradation rate of As(408). By reducing the TMAOH concentration to 0.83%, the conversion of the arsenosugar to As(328) and DMAA is nearly eliminated (less than 5% loss). Arsenosugars, As(408) and As(482), were also studied in 253 mM NaOH to verify the degradation products. The NaOH experiments were conducted to investigate a possible hydroxide based reaction mechanism. Similar degradation plots were found for each arsenosugar when compared to the 2.5% TMAOH data. A mechanism has been proposed for the formation of As(328) from As(408) and As(482) in base via an SN2 reaction (hydroxide attack) at the side chain carbon adjacent to the inorganic ester. The formation of DMAA is observed in all arsenosugars after prolonged exposure. This probably occurs via an SN2 attack at the arsenic atom. PMID- 14737233 TI - Determination of DNA by Rayleigh light scattering enhancement of molecular "light switches". AB - Base on the enhancement of Rayleigh light scattering signals of molecular "light switches" by DNA under acidic condition, a sensitive and convenient method for DNA determination was proposed. The experiments indicated that, under optimum conditions, good linear relationships were obtained between the Rayleigh light scattering intensity and the concentration of nucleic acids. The detect limits of calf thymus DNA (ctDNA) were 13.0 ng ml(-1), 4.2 ng ml(-1), 51.5 ng ml(-1) and 3.0 ng ml(-1) with four "light switches", respectively. Plasmid DNA extracted from Bacillus subtilis were determined by the proposed method with satisfactory results, and the recovery rates of calf thymus DNA were in the range of 94.6 110.7%. PMID- 14737234 TI - In-line monitoring of esterification using a miniaturised mid-infrared spectrometer. AB - A miniaturised mid-infrared (MIR) spectrometer attached directly to an in-line immersion probe with an attenuated total reflectance crystal has been used to monitor the progression of the batch reaction of crotonic acid and 2-butanol in toluene. Univariate calibration, using the signal at 1188 cm(-1) in the 2nd derivative spectrum, gave accurate (average error, 1.6%) and precise (average relative standard deviation, 5.2%) estimation of 2-butyl crotonate concentrations in the range 0.08-0.49 mol dm(-3). Calibration by partial least squares was of no additional benefit in this application. The performance of in-line MIR spectrometry was comparable to that of an off-line reference gas chromatography method and superior to that of other in-line process analysis techniques (near infrared, Raman or UV-visible spectrometries). PMID- 14737235 TI - Simultaneous determination of human serum albumin, gamma-globulin, and glucose in a phosphate buffer solution by near-infrared spectroscopy with moving window partial least-squares regression. AB - Near-infrared (NIR) spectra in the 12,000-4,000 cm(-1) region were measured for phosphate buffer solutions containing human serum albumin (HSA), gamma-globulin, and glucose with various concentrations at 37 degrees C. Five levels of full factorial design were used to prepare a sample set consisting of 125 samples of three component mixtures. The concentration ranges of HSA, gamma-globulin and glucose were 0.00-6.00 g dl(-1), 0.00-4.00 g dl(-1) and 0.00-2.00 g dl(-1), respectively. The 125 sample data were split into two sets, the calibration set with 95 data and the prediction set with 30 data. The most informative spectral ranges of 4648-4323, 4647-4255 and 4912-4304 cm(-1) were selected by moving window partial least-squares regression (MWPLSR) for HSA, [gamma]-globulin, and glucose in the mixtures, respectively. For HSA, the correlation coefficient (R) of 0.9998 and the root mean square error of prediction (RMSEP) of 0.0289 g dl(-1) were obtained. For [gamma]-globulin, R of 0.9997 and RMSEP of 0.0252 g dl(-1) were obtained. The corresponding statistic values of glucose were 0.9997 and 0.0156 g dl(-1), respectively. These statistical values obtained by MWPLSR are highly significant and better than those calculated by using the regions reported in the literature. The results presented here show that MWPLSR can select the informative regions with a simple procedure and increase the power of NIR spectroscopy for simultaneous determination of the concentrations of HSA, [gamma] globulin and glucose in the mixture systems. PMID- 14737236 TI - Microwave-assisted solvent extraction of the herbicide methabenzthiazuron from soils and some soil natural organic and inorganic constituents. Influence of environmental factors on its extractability. AB - A microwave-assisted solvent extraction (MASE) method for the determination of methabenzthiazuron (MBT) in soil samples by HPLC-DAD (diode array detection) was evaluated. Spiked soil samples having different physico-chemical properties, and selected soil-derived matrices with diverse MBT adsorption capacity, characterized by their Freundlich equation Kf values, were used to verify the method applicability to a broad range of different soils. The spiking procedure was considered a crucial point to reproduce as closely as possible the solute soil adsorption taking place in the natural environment. Ageing effects, where the compound could diffuse into inaccessible locations within the soil matrix in view of its great stability, were considered of particular concern. In spite of the heterogeneous physico-chemical properties of soils under study, recoveries were greater than 90%. Performance of the MASE procedure correlated highly with the adsorption capacity of soil-derived matrices: the lowest recoveries were for illite (67-73%), among the mineral surfaces, and for a humic acid (67-72%), among the organic fractions. Intra-assay variation for each type of sample soil range from 0.40 to 3.89%(RSD). Limits of detection and quantification were 0.047 and 0.15 microg g(-1), respectively. Analyte residence time was not a very significant factor on the extractability. PMID- 14737237 TI - An amobarbital molecularly imprinted microsphere for selective solid-phase extraction of phenobarbital from human urine and medicines and their determination by high-performance liquid chromatography. AB - A 40-60 microm amobarbital molecularly imprinted microsphere, used as a solid phase selective sorbent for extracting phenobarbital from human urine and medicines, was prepared by a suspension polymerization method. A series of binding studies was performed in order to find optimal loading, washing and eluting conditions for solid-phase extraction. Under optimal conditions, good recoveries of phenobarbital in samples were obtained. Normally, molecularly imprinted polymers, prepared in bulk, require laborious work. Significant losses occur during the procedure of grinding and washing. In this work all molecularly imprinted polymers made into microsphere could be utilized, and the cost of the template was reduced too (the price of phenobarbital is twice that of amobarbital). As the phenobarbital to be extracted was different from the template molecule amobarbital, the interference caused by template leaking could be avoided in the assay. PMID- 14737307 TI - Materials that naturally assemble themselves. AB - One of key aims of modern chemistry is to explore and exploit the phenomenal complexity of matter that seems to arise spontaneously within ambient environments like that of our planet. Biology, of course, is a ultimate manifestation of a universal chemical canon encompassing interactions both at the molecular and supramolecular level. Inorganic building blocks play a significant role in the self-organised assembly of many biological structures at various scale-lengths but the details of their chemical behaviour and interaction with organic compounds are still not well-understood. Stephen Mann of the University of Bristol is a pioneer in the area; his team is probing the ground rules for biomineralisation in the hope of applying them to the self-assembly of practical materials with complex hierarchical structures. PMID- 14737310 TI - Translocation of bioactive peptides across cell membranes by carbon nanotubes. AB - Functionalised carbon nanotubes are able to cross the cell membrane and to accumulate in the cytoplasm or reach the nucleus without being toxic for the cell up to 10 [micro sign]M. PMID- 14737309 TI - Synthetic hosts via molecular imprinting--are universal synthetic antibodies realistically possible? AB - Of the many ways to make synthetic hosts, one of the most appealing involves molecular imprinting. In the commonest approach monomer units assemble around or are attached to a template (imprint) molecule and then linked together using a cross-linking agent. Template removal ideally leaves cavities within the molecularly imprinted polymer (MIP) that possess a shape and functional group complementarity to the imprint molecule allowing its tight and selective uptake. This review highlights some recent advances in the synthesis of MIPs (often called "synthetic antibodies") and enumerates a "wish list" of properties for the perfect MIP that may guide future studies. PMID- 14737311 TI - Superstructures of donor packing arrangements in a series of molecular charge transfer salts. AB - Three conducting BEDT-TTF charge-transfer salts with tris(oxalato)metallate anions have unit cells containing both[small alpha] and [small beta][double prime] donor packing motifs. PMID- 14737312 TI - The initial stages of aminosilanol polymerisation. AB - For polysiloxanes to be used as a protective coating it is important that proton transfer, a trigger to polymerisation, is a facile process. Here we investigate the initial stages of polycondensation and compare different silanol tail groups and the effect of solvent (isopropanol). In the case of (3 aminopropyl)trihydroxysilane we see the potential for self catalysis as the tail group is a proton acceptor, while thiolpropyltrihydroxysilane and isopropanol do not promote proton transfer. PMID- 14737313 TI - The prediction of inorganic crystal framework structures using excluded regions within a genetic algorithm approach. AB - We have developed a new technique, which is complementary to other procedures, that will have wide applicability for generating new feasible framework structures with defined microporous architectures from the knowledge of only the unit cell dimensions, constituent elements and by defining forbidden regions within the unit cell. PMID- 14737314 TI - Controlled growth of calcium carbonate by poly(ethylenimine) at the air/water interface. AB - Two metastable calcium carbonate polymorphs, hemispherical vaterite and needle like aragonite, are selectively formed at the air/water interface by the mediation of poly(ethyleneimine)(with molecular weights of 25000 and 2000, respectively) dissolved in supersaturated calcium bicarbonate solution. PMID- 14737315 TI - Synthesis of carbon nitride nanotubes with the C(3)N(4) stoichiometry via a benzene-thermal process at low temperatures. AB - In this communication, we first report the direct synthesis of high-quality carbon nitride nanotubes (CNNTs) with inner diameters of 50-100 nm and wall thicknesses of 20-50 nm with the C(3)N(4) stoichiometry on a high-yield of 40%via a simple benzene-thermal process involving the reaction of C(3)N(3)Cl(3) with NaN(3) in a Teflon-lined autoclave at 220 [degree]C without using any catalyst or template. PMID- 14737316 TI - Self-assembly of a molecular figure-of-eight strip. AB - A hexanuclear copper(ii) complex with a figure-of-eight strip topology is formed by metal-directed self-assembly of tritopic ligand L, bis-bidentate glycine hydroxamic acid and Cu(ii) ions in a 2:2:6 ratio. PMID- 14737318 TI - A novel, tandem construction of C-N and C-C bonds: facile and one-pot transformation of the Baylis-Hillman adducts into 2-benzazepines. AB - A novel reaction involving tandem construction of C-N and C-C bonds via the simultaneous Ritter and Houben-Hoesch reactions on Baylis-Hillman adducts leading to a convenient, one-pot synthesis of 2-benzazepine derivatives is described. A facile stereoselective transformation of the Baylis-Hillman adducts into (E)- and (Z)-allyl amides is also presented. PMID- 14737317 TI - Layer-by-layer electrodeposition of redox polymers and enzymes on screen-printed carbon electrodes for the preparation of reagentless biosensors. AB - Layer-by-layer electrodeposition of redox polymer/enzyme composition films on screen-printed carbon electrodes for fabrication of reagentless enzyme biosensors has been proposed and the resulting films were found to be very stable and rigid. PMID- 14737319 TI - Preparation of hybrid thin film modified carbon nanotubes on glassy carbon electrode and its electrocatalysis for oxygen reduction. AB - A hybrid thin film containing Pt nanoparticles and [tetrakis(N methylpyridyl)porphyrinato]cobalt (CoTMPyP) modified multi-walled carbon nanotubes (MWNTs) on a glassy carbon (GC) electrode surface was fabricated. This hybrid film electrode exhibited remarkable electrocatalytic activity for oxygen reduction and high stability with promising applications in fuel cells. PMID- 14737320 TI - Renewable hydrogen by aqueous-phase reforming of glucose. AB - Hydrogen can be produced from aqueous solutions containing 10 wt% glucose with high selectivities through the combined use of a hydrogenation reactor for conversion of glucose to sorbitol, followed by a reforming reactor for conversion of sorbitol to H(2) and CO(2) and then a gas-liquid separator for the removal of high-pressure H(2)-rich reformate gas, ready for use in a fuel cell. PMID- 14737321 TI - Practical synthesis of new and highly efficient ligands for the Suzuki reaction of aryl chlorides. AB - A practical synthesis of a novel class of phosphine ligands, phosphino substituted N-aryl pyrroles (PAP ligands), has been developed. These ligands are applied in the palladium-catalyzed coupling of a variety of aryl and heteroaryl chlorides with phenylboronic acid showing exceedingly high turnover numbers at mild reaction temperatures and even at room temperature. PMID- 14737322 TI - Gas-phase reactions for selective detection of the explosives TNT and RDX. AB - Highly selective gas-phase reactions with ethyl vinyl ether (EVE) of major electron (EI) and chemical ionization (CI) fragment ions of the explosives TNT and RDX have been uncovered. The fragment ion of m/z 210 from TNT undergoes [4(+)+ 2] cycloaddition with EVE to form an oxo-iminium ion of m/z 282, which dissociates by acetaldehyde loss after a [1,5-H] shift to form a quinolynium ion of m/z 238. The fragment ion of m/z 149 from RDX reacts with EVE by a formal vinylation reaction, that is, the elusive cyclic adduct loses ethanol to yield a nitro-iminium ion of m/z 175, which reacts further with EVE to form a second cyclic product ion of m/z 247. Calculations and MS/MS experiments support the proposed structures. These highly characteristic reactions of diagnostic EI and CI fragment ions improve selectivity for TNT and RDX detection. PMID- 14737323 TI - Terminally functionalized polyisobutylene oligomers as soluble supports in catalysis. AB - A new phase selective hydrocarbon soluble polymer support is described. PMID- 14737324 TI - A new route to furanoeremophilane sesquiterpenoids. Synthesis of (+/-)-6[small beta]-hydroxyeuryopsin. AB - The naturally occurring furanoeremophilane 6[small beta]-hydroxyeuryopsin was synthesized by a novel route which involved Stille coupling of a 2-furylstannane with a cyclohexylmethyl bromide, followed by intramolecular formylation of the furan to complete the tricyclic nucleus of the sesquiterpenoid. PMID- 14737325 TI - Synthesis and characterization of large-pore vinyl-functionalized mesoporous silica SBA-15. AB - Ordered mesoporous silicas SBA-15 with high loadings of pendant vinyl groups have been synthesized via co-condensation of tetraethoxysilane (TEOS) and triethoxyvinylsilane (TEVS) templated with a triblock copolymer. PMID- 14737326 TI - Continuous synthesis of CdSe-ZnS composite nanoparticles in a microfluidic reactor. AB - ZnS-coated CdSe composite particles have been continuously synthesized in a microfluidic reactor. By using this system, CdSe particles and a ZnS coating can be produced in sequence, and the particle size and layer thickness can be directly adjusted by the residence time. It demonstrated that the continuous synthesis in the microreactor was a simple and efficient way to prepare composite particles with different structures and determine the optimized experimental conditions. PMID- 14737327 TI - Preparation and electrochemical behaviour of hydrophobic vitamin B(12) covalently immobilized onto platinum electrode. AB - Hydrophobic vitamin B(12) was covalently immobilized onto a platinum electrode surface, and the immobilized complex exhibits Co(ii)/Co(i) redox couple and in situ the Co(i) species reacts with phenethyl bromide to form styrene under irradiation with visible light with a turnover number of over 6000 for 1 h. PMID- 14737328 TI - Free energy of adsorption of water and calcium on the [10 1 4] calcite surface. AB - Molecular dynamics simulations of the calcite-water interface have shown that the free energy of adsorption of water is relatively small compared to the previously calculated enthalpy of adsorption implying a large entropy change and that the free energy profile of a calcium adsorbing on the surface correlates with the solvent density; these calculations allow us to begin to address the rates of adsorption and desorption which are essential for studying growth and dissolution. PMID- 14737329 TI - Very general formation of tetrahydropterin cation radicals during reaction of iron porphyrins with tetrahydropterins: model for the corresponding NO-synthase reaction. AB - Electron transfer from tetrahydropterins to iron porphyrins, with formation of intermediate tetrahydropterin cation radicals, is a very general reaction that was shown to occur not only with tetrahydrobiopterin, as originally found in NO synthases, but also with another important biological cofactor, tetrahydrofolate, and various iron porphyrins, either in their ferric state, or in the Fe(II)O(2) state, as in the first model of the corresponding NO-synthase reaction described in this paper. PMID- 14737330 TI - Synthesis of poly(para-phenylenevinylene) rotaxanes by aqueous Suzuki coupling. AB - PPV-based polyrotaxanes have been prepared by coupling vinyl boronic acids to aryl iodides in the presence of cyclodextrins, and the crystal structure of a [2]rotaxane of this type has been determined. PMID- 14737331 TI - Interactions between a surfactant and cavitand in water blur distinctions between host and guest. AB - The complexation of a water-soluble cavitand and sodium dodecyl sulfate micelles is studied using NMR diffusion ordered spectroscopy. PMID- 14737332 TI - Facile synthesis of a chiral polymeric helix; folding by intramolecular hydrogen bonding. AB - In a single condensation step, a poly-ureidophthalimide is synthesized, which folds into a chiral, helical architecture according to circular dichroism spectroscopy. PMID- 14737333 TI - Structural codons: linearity/helicity interconversion by pyridine/pyrimidine exchange in molecular strands. AB - Pyridine and pyrimidine groups connected through amide functions can be combined into specific sequences that self-organize into either helical or linear structures enforced by the formation of intramolecular hydrogen bonds. PMID- 14737334 TI - Selective 1,3-complexation of p-(t)Bu-calix[4]arene by [TiCp(2)Me(2)]. AB - Reaction of p-(t)Bu-calix[4]arene with dimethyl titanocene results in high yield selective 1,3-dimetallation of the calixarene in the cone conformation by selective cleavage of one methyl group. PMID- 14737335 TI - Disc-shaped triphenylenes in a smectic organisation. AB - A layered phase (SmA) was observed in a novel linked disc-rod mesogen, containing a triphenylene mesogen and attached to three cyanobiphenyl moieties. PMID- 14737336 TI - The role of borole in a fully conjugated electron-rich system. AB - The reaction of the 3,3[prime or minute]-dilithiobithieno complex with TipB(OMe)(2) affords a borole with an extended conjugated electron-rich [small pi]-electron system; the electronic perturbation by the introduction of push-pull substituents is described. PMID- 14737337 TI - Strong fluorescence emission induced by supramolecular assembly and gelation: luminescent organogel from nonemissive oxadiazole-based benzene-1,3,5 tricarboxamide gelator. AB - Supramolecular aggregation of a novel nonfluorescent gelator yields highly luminescent organogels in aprotic organic solvents through intermolecular hydrogen bonding, which is a key motif for both self-assembly and photophysical process control. PMID- 14737338 TI - Photochromism of triangle terthiophene derivatives as molecular re-router. AB - 2,2[prime or minute]-3,3[double prime]-Terthiophene derivatives undergo photochemically reversible cyclization and cycloreversion reactions. The absorption peak wavelength changed systematically with substitution of the phenyl rings at 5-, 5[prime or minute]- and 5[double prime]-positions of the thiophene rings, which indicates re-routing of the [small pi]-conjugation system. PMID- 14737339 TI - Thermally reversible fluorescent polymorphs of alkoxy-cyano-substituted diphenylbutadienes: role of crystal packing in solid state fluorescence. AB - Correlation of fluorescence and crystal packing in thermally interconvertible polymorphic states of octyloxy-cyano-substituted diphenylbutadiene possessing visually distinguishable fluorescence reveals that solid state fluorescence of this class of derivatives depends on their monomer-J-aggregate ratio, controlled by variations in their molecular packing. PMID- 14737340 TI - Relative importance of hydrogen bonding and coordinating groups in modulating the zinc-water acidity. AB - The presence of second-sphere -NH(2) groups in the proximity of a zinc(ii)-bound water molecule enhances its acidity by ca. 2 pK(a) units. PMID- 14737341 TI - Stable liquid crystalline phases of colloidally dispersed exfoliated layered niobates. AB - Colloidally dispersed niobium oxide nanosheets obtained by exfoliation of layered niobates HNb(3)O(8) and HTiNbO(5) formed stable liquid crystalline phases; their liquid crystallinity was dependent on the niobate species exfoliated. PMID- 14737342 TI - 3,6-dioxypyridazine bridged tungsten-tungsten quadruple bonds. Comparisons of electron delocalisation with oxalate bridged compounds. AB - The preparation and characterisation of the tungsten-tungsten quadruply bonded, 3,6-dioxypyridazine bridged complex [((t)BuCO(2))(3)W(2)](2)([micro sign] H(2)C(4)N(2)O(2)) and its single electron oxidised radical cation are reported and, when compared with related bridged dimolybdenum complexes, reveal a different mechanism of electronic coupling from that seen in related oxalate bridged systems. PMID- 14737343 TI - Novel ambient light induced trimerization of a simple copper(II) monomeric Schiff base complex. AB - Room temperature, light promoted trimerization of a monomeric complex [Cu(HL)(L)](NO(3)).H(2)O (HL =N,N-dimethyl-N[prime or minute] propylsalicylideneimine) dissolved in CH(2)Cl(2), was observed with the concomitant generation of new ligand moiety (ClCH(2)L). PMID- 14737344 TI - Carbon-carbon bond cleavage by cytochrome p450(BioI)(CYP107H1). AB - Cytochrome p450(BioI)(CYP107H1) is believed to supply pimelic acid equivalents for biotin biosynthesis in Bacillus subtilis: we report here that the mechanistic pathway adopted by this multifunctional p450 for the in-chain cleavage of fatty acids is via consecutive formation of alcohol and threo-diol intermediates, with the likely absolute configuration of the intermediates also reported. PMID- 14737345 TI - Blue-shifted lithium bonds. AB - A number of lithium bonding systems (X-LiY) have been found in which the X-Li bond is shortened due to the lithium bond formation. PMID- 14737346 TI - Borrowing hydrogen: a catalytic route to C-C bond formation from alcohols. AB - Ruthenium complexes have been shown to perform efficient transfer hydrogenation reactions between alcohols and alkenes; in combination with an in situ Wittig reaction, indirect formation of C-C bonds has been achieved from alcohols. PMID- 14737347 TI - A general entry to linear, dendritic and branched thiourea-linked glycooligomers as new motifs for phosphate ester recognition in water. AB - A blockwise iterative synthetic strategy for the preparation of linear, dendritic and branched full-carbohydrate architectures has been developed by using sugar azido(carbamate) isothiocyanates as key templates; the presence of intersaccharide thiourea bridges provides anchoring points for hydrogen bond directed molecular recognition of phosphate esters in water. PMID- 14737349 TI - Electrostatically arranged cytochrome c-fullerene photoelectrodes. AB - We have developed a molecular-level switch-a C(60)/cytochrome c modified ITO electrode-that reversibly transmits and processes solar energy. PMID- 14737348 TI - Switching extended 1,3-diequatorial and bent 1,3-diaxial states of a disubstituted hinge sugar by ligand exchange reactions on Pt(II). AB - The bent conformation of a trisaccharide containing 2,4-diaminoxylopyranoside, in which both end sugars are presented in 1,3-diaxial orientation, is fixed by chelation of the diamino groups to Pt(II) and unfixed by a ligand exchange reaction with NaCN or thiourea giving its extended conformation. PMID- 14737350 TI - Highly enantioselective spiro cyclization of 1,6-enynes catalyzed by cationic skewphos rhodium(I) complex. AB - A cationic rhodium(I) complex having a skewphos ligand is shown to be a highly enantioselective catalyst for asymmetric carbocyclization of 1,6-enynes with tri substituted olefins to control quaternary stereogenic centers of spiro-rings. PMID- 14737351 TI - Large macroscopic size changes in chemomechanical polymers with binding sites for metal ions. AB - A flexible polymer based on the reaction of polyacrylate with diethylenetriamine shows, under the control of transition metal ions in the external aqueous medium, size expansion or contraction by up to 200%(corresponding to 400% volume change); the macroscopic movements, which are fully reversible, show a distinct maximum at a certain metal salt concentration, and vary significantly with the addition of second effector compounds such as sodium chloride or benzoate. PMID- 14737352 TI - First synthesis of an amythiamicin pyridine cluster. AB - The pyridine-containing central domain of the amythiamicin group of thiopeptide antibiotics is prepared in protected form in 9 steps, 93%ee and 18% overall yield from (S)-2-[1-(tert-butoxycarbonylamino)-2-methylpropyl]thiazole-4-carboxylic acid by Michael addition-cyclodehydration of a 2-(2-thiazolyl)enamine and 1-(2 thiazolyl)propynone. PMID- 14737353 TI - Facile palladium-catalysed synthesis of 1-aryl-1H-indazoles from 2 bromobenzaldehydes and arylhydrazines. AB - 2-Bromobenzaldehydes react with arylhydrazines in toluene at 100 [degree]C in the presence of a catalytic amount of a palladium catalyst and phosphorus chelating ligands such as 1,1[prime or minute]-bis(diphenylphosphino)ferrocene and 1,3 bis(diphenylphosphino)propane along with NaO-t-Bu to afford 1-aryl-1H-indazoles in good yields. PMID- 14737354 TI - Preparation and structure of 2-iodoxybenzoate esters: soluble and stable periodinane oxidizing reagents. AB - Esters of 2-iodoxybenzoic acid (IBX-esters) are potentially valuable oxidizing reagents belonging to a new class of pentavalent iodine compounds with a pseudo benziodoxole structure. PMID- 14737355 TI - Photomodulated molecular recognition of the guanidinium cation. AB - Azobenzene moieties were incorporated into a synthetic receptor allowing its affinity for the guanidinium cation to be modulated ten-fold by photoirradiation and/or heating. PMID- 14737356 TI - Isolation and structural characterization of the first thermally robust and air stable Cr(4+) bent-metallocene complex. AB - The first thermally robust and air stable bent-sandwich chromocene complex with chromium in the +4 oxidation state has been isolated and fully characterized. PMID- 14737357 TI - Reversible carboxylation of N-heterocyclic carbenes. AB - Spectroscopic analysis, thermogravimetric analysis, and crossover experiments performed on a series of imidazolium carboxylates revealed carboxylation was reversible with N-aryl substituted adducts. PMID- 14737358 TI - Directed regio- and stereoselective hydroformylation of mono- and 1,3 disubstituted allylic alcohols: a catalytic approach to the anti-aldol-retron. AB - Regioselective and diastereoselective hydroformylation of mono- and 1,3 disubstituted allylic alcohol o-DPPB esters is described. The products represent synthetically important anti-aldol retrons. PMID- 14737359 TI - Regio- and enantioselective iridium-catalysed allylic aminations and alkylations of dienyl esters. AB - Regio- and enantioselective iridium-catalysed allylic aminations and alkylations of dienyl substrates are presented; using phosphorus amidite L1 as ligand, aminations provided ee values of up to 97% and alkylations of up to 90%. PMID- 14737360 TI - The first SERRS multiplexing from labelled oligonucleotides in a microfluidics lab-on-a-chip. AB - The first simultaneous detection of three dye-labelled oligonucleotides in a microfluidics chip by SERRS is reported. PMID- 14737361 TI - Formation and spectroscopic characterization of the dioxygen adduct of a heme-Cu complex possessing a cross-linked tyrosine-histidine mimic: modeling the active site of cytochrome c oxidase. AB - A binucleating porphyrin with covalently appended copper chelates having a cross linked imidazole-phenol group as the novel active site model of cytochrome c oxidase has been prepared, and the dioxygen adduct of its iron(II)-copper(I) complex was spectroscopically characterized. PMID- 14737362 TI - A new synthesis of 1,3-dihydrobenzo[1,2]oxasiloles by a novel rearrangement of a pentavalent silicon intermediate. AB - A new synthesis of benzo[1,2]oxasiloles is described, wherein an unprecedented intramolecular allylic transposition takes place probably involving a pentavalent silicon intermediate. PMID- 14737363 TI - Novel and efficient synthesis of difficult sequence-containing peptides through O N intramolecular acyl migration reaction of O-acyl isopeptides. AB - A novel and efficient method for the synthesis of difficult sequence-containing peptides has been developed based on the synthesis of O-acyl isopeptides followed by an O-N intramolecular acyl migration reaction, resulting in a remarkable improvement of the yields. PMID- 14737461 TI - Airborne sulfur and nitrogen in Finland-trends and exposure in relation to air transport sector. AB - We present the concentration trends and the atmospheric exposure of sulfur and nitrogen compounds in different transport sectors during the period 1981-2000, based on the air quality data of the Finnish Meteorological Institute. Sulfur and nitrogen concentrations in air and precipitation were assessed from background stations covering the whole country. A significant decrease of more than 60% was observed in the concentrations of all sulfur compounds throughout the country during 1981-2000. In the 1990's, significant trends were detected in all sulfur time series, with the exception of sulfate in precipitation at the northernmost stations. On the other hand, the concentrations of oxidized nitrogen compounds in air and precipitation have declined only slightly or not at all, especially in the 1990's. This is attributed to the fairly small domestic and European NO(x) emission reductions after 1981. The ammonium concentration in precipitation has declined by more than 50% during 1981-2000. The atmospheric sulfur exposure was found to be dominated by transport from the West and South-West sectors during summer, whereas in winter all the southward sectors were important. The largest decline in the sectoral sulfur concentrations has taken place already before 1991, with a more pronounced reduction during winter than during summer. The atmospheric nitrogen exposure was mainly dominated by transport from the sectors between west and south, but in the latter half of the study period there seems to have been a slight shift towards increased transport from the eastern and northern sectors. PMID- 14737464 TI - Evaluation of method of preparation of passive diffusion tubes for measurement of ambient nitrogen dioxide. AB - This study was carried out in response to suggestions that the measurement of NO(2) by Palmes-type passive diffusion tubes (PDT) is affected by the method of preparation of the triethanolamine (TEA) absorbent coating on the grids. The following combinations of factors were investigated: TEA solvent (acetone or water), volume composition of TEA in solvent (50% or 20%), and grid coating method (dipping in solution prior to assembly or pipetting solution on after assembly). Duplicate PDTs prepared by each of the 8 methods were exposed in parallel, in urban air, for a total of 80 separate 1 week exposures. NO(2) concentrations derived from PDTs prepared by pipetting methods were significantly less precise than concentrations from dipping methods, with mean RSDs for duplicate measurements of 13.8% and 8.5%, respectively (n= 316 each category). Pipetting methods using solutions of 50% TEA composition were particularly imprecise (mean RSD 17.2%). Data from PDTs prepared by pipetting methods were systematically more poorly correlated with each other and with data from co located chemiluminescence analysers, than corresponding data from PDTs prepared by dipping methods, indicating that more consistent accuracy was also obtained by the latter PDTs. The statistical evidence suggested that PDTs prepared by pipetting 50% TEA in water generally gave lower NO(2) concentrations. Although this is in agreement with a previous study, it is also possible that such an observation here may be a statistical artefact given the demonstrably poorer precision of this method. The general tendency of PDTs to show positive bias in NO(2) measurement in urban air in 1 week exposures was again evident in this study (mean biases at roadside and urban centre locations of +35% (n= 475) and +18% (n= 112), respectively) consistent with augmentation of within-tube NO(2) flux by chemical reaction between co-diffusing NO and O(3). Overall, it is recommended that the pipetting method of PDT grid preparation is avoided, or at least investigated further, because of the apparent degradation in precision and accuracy of NO(2) measurement. Potential reasons for the effect are discussed. PMID- 14737465 TI - Comparison of wood-dust aerosol size-distributions collected by air samplers. AB - A method has been described previously for determining particle size distributions in the inhalable size range collected by personal samplers for wood dust. In this method, the particles collected by a sampler are removed, suspended, and re-deposited on a mixed cellulose-ester filter, and examined by optical microscopy to determine particle aerodynamic diameters. This method is particularly appropriate to wood-dust particles which are generally large and close to rectangular prisms in shape. The method was used to investigate the differences in total mass found previously in studies of side-by-side sample collection with different sampler types. Over 200 wood-dust samples were collected in three different wood-products industries, using the traditional 37 mm closed-face polystyrene/acrylonitrile cassette (CFC), the Institute of Occupational Medicine (IOM) inhalable sampler, and the Button sampler developed by the University of Cincinnati. Total mass concentration results from the samplers were found to be in approximately the same ratio as those from traditional long-term gravimetric samples, but about an order of magnitude higher. Investigation of the size distributions revealed several differences between the samplers. The wood dust particulate mass appears to be concentrated in the range 10-70 aerodynamic equivalent diameter (AED), but with a substantial mass contribution from particles larger than 100 microm AED in a significant number of samples. These ultra-large particles were found in 65% of the IOM samples, 42% of the CFC samples and 32% of the Button samples. Where present, particles of this size range dominated the total mass collected, contributing an average 53% (range 10-95%). However, significant differences were still found after removal of the ultra-large particles. In general, the IOM and CFC samplers appeared to operate in accordance with previous laboratory studies, such that they both collected similar quantities of particles at the smaller diameters, up to about 30-40 [micro sign]m AED, after which the CFC collection efficiency was reduced dramatically compared to the IOM. The Button sampler collected significantly less than the IOM at particle sizes between 10.1 and 50 microm AED. The collection efficiency of the Button sampler was significantly different from that of the CFC for particle sizes between 10.1 and 40 microm AED, and the total mass concentration given by the Button sampler was significantly less than that given by the CFC, even in the absence of ultra-large particles. The results are consistent with some relevant laboratory studies. PMID- 14737466 TI - Arsenic biomonitoring using a hyperaccumulator fern (Pteris vittata). AB - Samples of Pteris vittata L. (brake fern or ladder brake) collected in Genova and in areas outside urban centres, have been analysed for arsenic content in order to assess if hyper accumulating plants are suitable for monitoring purposes. Hyper accumulation ability of the Ligurian fern populations was evaluated by analysing specimens grown with hydroponic media added with As(v). Arsenic concentrations in the range 2-310 microg g(-1) dry weight have been measured in samples collected in different sites along the Ligurian coast. Arsenic concentrations in fern fronds correlate with the estimated arsenic emission in the area, verifying the applicability of P. vittata as an arsenic biomonitor. PMID- 14737467 TI - An inventory-based carbon budget for forest and woodland ecosystems of Turkey. AB - Environmental monitoring of national-level comparisons of CO(2) emissions is needed to quantify sources and sinks of carbon (C) in national ecosystems. In this study, a national forest inventory database was used to estimate the past and current pools and fluxes of C in deciduous and coniferous forest and woodland ecosystems (20.7 x 10(6) ha) of Turkey. Growing C stock was 12.63 t C ha(-1) in 1960 and 16.55 t C ha(-1) in 1995. Total C store in the whole live woody biomass was estimated at 22.77 t C ha(-1) in 1996. The total flux of C from the atmosphere into the forest and woodland ecosystems driven by primary productivity was about 1.46 t C ha(-1)(or 30.2 Mt C) in 1996. The estimated net release of C from the forest and woodland ecosystems of Turkey to the atmosphere was about 1.34 t C ha(-1)(or 21.5 Mt C) in 1996. When C released was taken into account, net ecosystem sequestration (NES) resulted in 0.12 t C ha(-1) per year. Such analytical tools as national forest C budgets are needed to improve our preventive and mitigative strategies for dealing with global climate change. PMID- 14737468 TI - A study of the elemental leachability and retention capability of compost. AB - In this work a comparison is made between the different approaches that can be taken to evaluate the mobility of elements in compost. The practical consequences of the results obtained are also discussed in terms of methods for cleaning up compost and using compost in environmental remediation. The mobility of potentially toxic elements in compost is evaluated by leaching with four selected eluents, i.e. diluted sulfuric acid, oxalate, citrate and EDTA. In contrast to the chelating agents, diluted sulfuric acid was found to generally have a low leaching capability for removal of heavy metals from compost. This implies that the risk of heavy metal leaching caused by natural rainfall is likely to be low. The results obtained in the leaching experiment were compared with previous results obtained from sequential fractionation. This comparison confirmed that both methods gave similar results for predicting the lability of elements in compost. A non-linear regression analysis of the leaching curves was also conducted. The leaching curves for elements with high lability could be fitted with a two components model. The labile components identified by the kinetic model are approximately in accordance with the fractions obtained from the first step of the sequential extraction method. The kinetic speciation method is shown to be a relatively rapid and simple procedure for compost which gives more information about element lability than simple leaching experiments. The leaching reagents used in this work were not effective enough to be used for cleaning up compost with a high metal content. Compost was however shown to have a high affinity for heavy metals, with the order of affinity of metal for the compost being very similar to that seen for humic acid. Compost may therefore prove to be a good remediation material for metal contaminated waste. PMID- 14737469 TI - Extractable copper, arsenic and antimony by EDTA solution from agricultural Chilean soils and its transfer to alfalfa plants (Medicago sativa L.). AB - Following our research on copper, arsenic and antimony in Chilean ecosystems, a study to understand the mobility and transport of these elements from soil to plants was carried out. So, the aim of this study, which follows on from the previous work, was to demonstrate if the total concentrations of these elements or their fractions extracted by 0.05 M EDTA pH 7 from different Chilean soils correlate with the respective total concentrations in the edible tissue of alfalfa plants collected simultaneously from 20 different sites affected or unaffected by mining activities. The highest copper fractions extracted by EDTA solutions were obtained in contaminated soils from the central region (41-69%); however the northern soils presented the highest extractable fractions of arsenic (9-34%). The antimony fraction was low in all soils (0.4-8.0%). Alfalfa plants from all contaminated sites presented high copper, arsenic and antimony concentrations (19-126 mg kg(-1), 5.7-16.3 mg kg(-1) and 0.16-1.7 mg kg(-1), respectively). Statistically significant correlations were obtained between the total contents of copper and arsenic and their respective extractable fractions in soils. Good correlations were found between elements in alfalfa plants. Correlations were also obtained between the total concentrations of three elements in soils and in alfalfa plants. However, excepting for antimony in the northern samples, higher correlation coefficients were evaluated when the extractable fractions were considered. Samples from the north region presented the highest copper transfer factor and the lowest for arsenic, in spite of the high concentration of this metalloid extracted by EDTA solution in these soils. There was not a clear trend on the transfer factor for antimony, probably due to the low content of this element in alfalfa plants and/or the low recovery obtained for this element by EDTA. PMID- 14737470 TI - Characterisation of trace elements and methylmercury in an estuarine sediment reference material, IAEA-405. AB - An estuarine sediment sample, IAEA-405, was recently characterised for trace elements and methylmercury (MeHg) for ultimate use as a marine reference material. The reference values were calculated using data sets from 120 laboratories that participated in a world-wide intercomparison exercise. The data evaluation is highlighted, and includes comparisons of different analytical methods, as well as the distribution of data. Overall laboratory performance for this exercise was encouraging: 86 laboratories (72%) reported data with <25% outliers. There was sufficient acceptable data to establish recommended values for 17 elements and information values for a further 15 elements. In addition, a recommended value was established for methylmercury. The estuarine sediment sample, IAEA-405, can be used as a reference material for quality control in the determination of trace elements and methylmercury in moderately polluted sediments. PMID- 14737471 TI - The fate of fipronil in modular estuarine mesocosms. AB - The degradation and corresponding product manifold for the pesticide fipronil was determined in three replicate estuarine mesocosms. Aqueous fipronil concentrations rapidly decreased over the 672 h timescale of the experiment (95% removal). Loss was apparently first-order in fipronil, although there appeared to be a change in the removal mechanism after 96 h that corresponded to a dramatic slowdown in its disappearance. The reduction product of fipronil, fipronil sulfide, was not detected in the water column; however, it formed rapidly in sediments and was identified as the major product of fipronil degradation in the system (20% yield at 672 h, with respect to initial fipronil concentration). Fipronil sulfone is thought to form primarily via biological oxidation; and, although it was generated rapidly in the water column (10% yield), only trace amounts were detected in the sediment (1% yield). The direct photolysis product of fipronil, fipronil desulfinyl, was present in all samples; it formed rapidly in the water column (4% yield) and partitioned into the sediment phase (7% yield) over the course of the experiment. The mass balance on fipronil and associated products was 42% at 672 h. PMID- 14737472 TI - Quantification of bacterial lipopolysaccharides (endotoxin) by GC-MS determination of 3-hydroxy fatty acids. AB - A GC-MS method for the quantification of bacterial lipopolysaccharides (LPS, endotoxin) is presented. After hydrolytic cleavage of 3-hydroxy fatty acids (3-OH FAs) from the lipid A region of LPS, derivatisation of both the hydroxyl and the carboxyl group was performed in one step with a mixture of methyl bis(trifluoracetamide)(MBTFA) and N-methyl-N-(tert butyldimethylsilyl)trifluoracetamide (MTBSTFA). Using GC-MS in the EI mode with selected ion monitoring (SIM) for analysis, baseline separation of 3-OH FAs (and of possibly interfering 2-OH FAs) was achieved. The sensitivity of the method (LOD 7-50 pg/injection for the different 3-OH FAs investigated) allows for the efficient quantification of LPS in occupational and environmental samples. Degradation of 3-OH FAs as well as of their derivatives during sample preparation and GC-MS separation as a possible source of errors in analytical methods based on 3-OH FA determination is reported for the first time. Thermal elimination of water from the underivatised 3-OH FAs and of trifluoroacetic acid from the derivatives was identified as the cause of degradation. The resulting alpha,beta unsaturated compounds showing the same mass spectra as the 3-OH FA derivatives were detected as more or less prominent satellite peaks. By using alkaline instead of acidic hydrolysis and cool on-column instead of split/splitless injection, elimination was reduced to an acceptable level. PMID- 14737473 TI - A yeast-based cytotoxicity and genotoxicity assay for environmental monitoring using novel portable instrumentation. AB - An assay capable of simultaneously measuring both general toxicity and more subtle genotoxicity, in aqueous environmental samples, is described. The assay uses eukaryotic (yeast) cells, genetically modified to express a green fluorescent protein (GFP) whenever DNA damage, as a result of exposure to genotoxic agents, is repaired. A measure of the reduction in cell proliferation is used to characterise general toxicity producing familiar EC(50) and LOEC data. The assay protocol has been developed for proposed use in the field and hence employs dedicated, portable instrumentation, the development of which is described. A range of environmentally relevant substances has been evaluated using the assay, including solutions of metal ions, solvents and pesticides. Preliminary data comparing the yeast assay's response to that of a standard Daphnia test in the analysis of the toxicity of 34 varied industrial waste effluents are also presented. The sensitivity to a wide range of substances and effluents suggests the assay should be useful for environmental toxicity monitoring. PMID- 14737474 TI - Analysis of BTEX and other substituted benzenes in water using headspace SPME-GC FID: method validation. AB - The analysis of BTEX and other substituted benzenes in water samples using solid phase microextraction (SPME) and quantification by gas chromatography with flame ionization detection (GC-FID) was validated. The best analytical conditions were obtained using PDMS/DVB/CAR fibre using headspace extraction (HS-SPME) at 50 [degree]C for 20 min without stirring. The linear range for each compound by HS SPME with GC/FID was defined. The detection limits for these compounds obtained with PDMS/DVB/CAR fibre and GC/FID were: benzene (15 ng L(-1)), toluene (160 ng L(-1)), monochlorobenzene (54 ng L(-1)), ethylbenzene (32 ng L(-1)), m-xylene (56 ng L(-1)), p-xylene (69 ng L(-1)), styrene (35 ng L(-1)), o-xylene (42 ng L(-1)), m-dichlorobenzene (180 ng L(-1)), p-dichlorobenzene (230 ng L(-1)), o dichlorobenzene (250 ng L(-1)) and trichlorobenzene (260 ng L(-1)). This headspace SPME-GC-FID method was compared with a previously validated method of analysis using closed-loop-stripping analysis (CLSA). The headspace SPME-GC-FID method is suitable for monitoring the production and distribution of potable water and was used, in field trials, for the analysis of samples from main intakes of water (surface or underground) and from the water supply system of a large area (Lisbon and neighbouring municipalities). PMID- 14737475 TI - Effects of organic ligands and pH on the leaching of copper from brake wear debris in model environmental solutions. AB - Copper leaching from a disc brake wear debris sample was examined in a variety of aqueous solutions to simulate potential leaching processes during rain events and in surface waters. Synthetic rainwater leached 40% of the total copper present in the brake wear debris into solution after 18 h in batch reactors, which was approximately three times more copper than that extracted by the US Environmental Protection Agency's Synthetic Precipitation Leaching Procedure. Formate and acetate were responsible for the enhanced copper leaching, as demonstrated by higher average amounts of leached copper in synthetic rainwater with- versus without the organic acids (40 versus 31% recovery). This observation suggests leaching tests that do not incorporate the appropriate types and concentrations of organic ligands present in rainwater will likely underestimate copper mobilization from brake wear debris during rain events. Leaching of copper from the brake wear debris ranged from 23 to 40% in solutions containing 3 to 15 mg C L(-1) dissolved humic substances, and was higher still in solutions containing relatively high concentrations of the synthetic metal chelating agent ethylenediaminetetraacetic acid. Static pH tests demonstrated that copper leaching from brake wear debris is highly pH dependent, with more leaching occurring at lower solution pH values. Leaching rate studies revealed that equilibrium generally was not attained within 48 h in the model solutions, indicating that additional copper can be expected to be released in environments where brake wear debris is exposed to long-term leaching processes. PMID- 14737505 TI - Polymer brushes via surface-initiated polymerizations. AB - Polymer brushes produced by controlled surface-initiated polymerization provide a route to surfaces coated with well-defined thin polymer films that are covalently bound to the substrate. All of the major controlled polymerization techniques have been applied to the synthesis of polymer brushes and examples of each are presented here. Many examples of brush synthesis in the literature have used the living atom transfer radical polymerization (ATRP) system, and in this tutorial review a particular focus is given to examples of this technique. PMID- 14737504 TI - Sports drug testing--an analyst's perspective. AB - Sport plays a major role in the lives of many people, both for active participation and as entertainment. Sport is now a huge nationally and internationally based industry. The desire to win has led some athletes to resort to the use of performance enhancing drugs. With huge financial rewards now available in some sports the pressure to excel has grown. Some have argued that drug use should be given free rein, however most people are of the view that it is athletic prowess that should be applauded not the efficacy of various performance enhancing drugs. Apart from the obvious aspects of equality and fair play, the use of drugs is associated with significant health risks. In the 1960's the use of stimulants in sports such as cycling led to the death of at least one cyclist. Since 1968 the International Olympic Committee (IOC) has required all Olympic Games' host cities to provide laboratory facilities for the analysis and detection of performance enhancing drugs. There are now 29 IOC accredited laboratories throughout the world that routinely test samples from athletes for the presence of such drugs. The purpose of this tutorial review is to give an overview of drug testing procedures, including those that were used at the last summer Olympic Games in Sydney 2000, and the incorporation of the latest developments in analytical chemistry technology in the drug testing process. More recently, developments in biotechnology mean that the use of whole new classes of drugs are banned in sport, often requiring new methodologies and techniques for their analysis. The contest between those who wish to cheat and those who wish to maintain fair play in sport is an ongoing one. PMID- 14737506 TI - Pulsed laser ablation and deposition of thin films. AB - Pulsed laser ablation is a simple, but versatile, experimental method that finds use as a means of patterning a very diverse range of materials, and in wide areas of thin film deposition and multi-layer research. Superficially, at least, the technique is conceptually simple also, but this apparent simplicity hides a wealth of fascinating, and still incompletely understood, chemical physics. This overview traces our current physico-chemical understanding of the evolution of material from target ablation through to the deposited film, addressing the initial laser-target interactions by which solid material enters the gas phase, the processing and propagation of material in the plume of ejected material, and the eventual accommodation of gas phase species onto the substrate that is to be coated. It is intended that this Review be of interest both to materials scientists interested in thin film growth, and to chemical physicists whose primary interest is with more fundamental aspects of the processes of pulsed laser ablation and deposition. PMID- 14737507 TI - The Pauson-Khand reaction, a powerful synthetic tool for the synthesis of complex molecules. AB - There are still some synthetic chemists who hesitate to use metal-mediated or catalysed reactions. The Pauson-Khand reaction (PKR) is a powerful transformation that has now been sufficiently well developed to be routinely considered when planning a synthesis, especially of polycyclic complex molecules. This tutorial review aims to encourage the use of this process explaining the best ways of performing a PKR both in the stoichiometric and the catalytic version, showing the scope of the process and its limitations. Additionally, asymmetry can be introduced in the reaction using several strategies, which will be discussed. The most recent examples of the synthetic applications of the PKR in natural product synthesis will give the reader an idea of the great usefulness of this reaction. PMID- 14737515 TI - Atropisomers and near-atropisomers: achieving stereoselectivity by exploiting the conformational preferences of aromatic amides. AB - The conformational preferences of aromatic amides are remarkably easy to control with a high degree of selectivity. This article reviews the consequences of this unusual form of stereocontrol, which enables for example the asymmetric synthesis of atropisomers and the ability to achieve remote stereocontrol by conformational relay. PMID- 14737508 TI - Dendrimers in drug research. AB - Dendrimers are versatile, derivatisable, well-defined, compartmentalised chemical polymers with sizes and physicochemical properties resembling those of biomolecules e.g. proteins. The present critical review (citing 158 references) briefly describes dendrimer design, nomenclature and divergent/convergent dendrimer synthesis. The characteristic physicochemical features of dendrimers are highlighted, showing the effect of solvent pH and polarity on their spatial structure. The use of dendrimers in biological systems are reviewed, with emphasis on the biocompatibility of dendrimers, such as in vitro and in vivo cytotoxicity, as well as biopermeability, biostability and immunogenicity. The review deals with numerous applications of dendrimers as tools for efficient multivalent presentation of biological ligands in biospecific recognition, inhibition and targeting. Dendrimers may be used as drugs for antibacterial and antiviral treatment and have found use as antitumor agents. The review highlights the use of dendrimers as drug or gene delivery devices in e.g. anticancer therapy, and the design of different host-guest binding motifs directed towards medical applications is described. Other specific examples are the use of dendrimers as 'glycocarriers' for the controlled multimeric presentation of biologically relevant carbohydrate moieties which are useful for targeting modified tissue in malignant diseases for diagnostic and therapeutic purposes. Finally, the use of specific types of dendrimers as scaffolds for presenting vaccine antigens, especially peptides, for use in vaccines is presented. PMID- 14737516 TI - Polymeric silver(I) coordination tubes. AB - Isolated polymeric Ag(I) coordination tubes are self-assembled from the rigid triamino ligands cis,cis-1,3,5-triaminocyclohexane (cis-tach) and cis,trans-1,3,5 triaminocyclohexane (trans-tach), forming two topologically equivalent framework motifs. PMID- 14737517 TI - Host-guest interactions template: the synthesis of a [3]catenane. AB - Formation of a [3]catenane containing dibenzo-24-crown ether wheels and a large dipyridiniumethane ring is templated by formation of a host-guest adduct between the [3]catenane and the external crown ether. PMID- 14737518 TI - First Br4 four centre-six electron and Se2Br5 seven centre-ten electron bonds in nonionic bromine adducts of selenanthrene. AB - Extended hypervalent Br(4) 4c-6e and Se(2)Br(5) 7c-10e bonds are detected in nonionic bromine adducts of selenanthrene, where the four Br(4) and seven Se(2)Br(5) atoms align linearly. PMID- 14737519 TI - Phosphonate ligands encourage a platonic relationship between cobalt(II) and alkali metal ions. AB - Two new cobalt(ii) cages are reported where the metal core has a high symmetry related to a Platonic solid; the choice of alkali metal used in the base used for deprotonation appears to influence the resulting structures. PMID- 14737520 TI - Poly(2,7-di-n-pentyldibenzofulvene) showing chiroptical properties in the solid state based purely on a chiral conformation. AB - Poly(2,7-di-n-pentyldibenzofulvene)s with no stereocenters, obtained by anionic polymerization using chiral 9-fluorenyllithium ligand complexes, showed intense circular dichroism in film form due purely to the chiral conformation of the main chain, while no chiroptical properties were detectable in solution. PMID- 14737521 TI - Cycloaddition of phosphanylidene-sigma4-phosphoranes ArP=PMe3 and quinones to yield 1,3,2-dioxophospholanes. AB - The reaction of phosphanylidene-[sigma](4)-phosphoranes ArP=PMe(3)(Ar = 2,6 Mes(2)C(6)H(3) or 2,4,6-t-Bu(3)C(6)H(2)) with select ortho-quinones yields 1,3,2 dioxophospholanes, one of which shows interesting [small pi]-stacking of the aromatic groups in the solid state. PMID- 14737522 TI - The structure of poly(ethylene oxide)8: NaBPh4 from a single crystal oligomer and polycrystalline polymer. AB - We show that the structure of a polymer electrolyte may be solved by growing single crystals of an oligomeric (short chain) complex which provided an adequate starting model for refinement of the equivalent polymeric structure using powder diffraction: the efficacy of this method has been demonstrated by determining for the first time the structure of an 8 : 1 complex, poly(ethylene oxide)(8) : NaBPh(4). PMID- 14737523 TI - NMR and computational studies of the chemical reduction of [2.2]paracyclophane: formation of dianionic p-xylenyl oligomers. AB - Reaction of [2.2]paracyclophane with K/Na alloy in THF gives a p-xylylenyl dianion together with its dimer and trimer which are relatively stable at low temperatures; at ambient temperatures further polymerization takes place. PMID- 14737524 TI - Inherent helicity in an extended tris-bipyridyl molecular cage. AB - A new molecular cage incorporating three bipyridyl units has been synthesised by a conventional multi-step procedure as well as, much more efficiently, by a Ni(ii) template procedure; an X-ray structure of the nickel complex shows that it adopts an exo configuration of each of the bridgehead nitrogen lone pairs, the central metal ion acts to promote a triple helical twist that extends [similar]22 [Angstrom] along the axial length of the molecule. PMID- 14737525 TI - Synthesis of new thermotropic liquid crystalline polyurethanes containing biphenyl mesogens using a novel AB-type self-polycondensation. AB - A series of thermotropic main chain liquid crystalline polyurethanes containing biphenyl mesogens and flexible methylene spacers were synthesized using the novel AB-type self-polycondensation approach for the first time. PMID- 14737526 TI - Structure and magnetism of a new pyrazolate bridged iron(II) spin crossover complex displaying a single HS-HS to LS-LS transition. AB - The dinuclear iron(II) complex [(pypzH)(NCSe)Fe([micro sign] pypz)(2)Fe(NCSe)(pypzH)].2H(2)O displays a single, sharp spin crossover transition between the [HS-HS] and [LS-LS] states and is structurally characterised above and below the T(1/2)= 225 K value PMID- 14737527 TI - The regiospecific Fischer indole reaction in choline chloride.2ZnCl2 with product isolation by direct sublimation from the ionic liquid. AB - The Fischer indole synthesis occurs in high yield with one equivalent of the ionic liquid choline chloride[middle dot]2ZnCl(2); exclusive formation of 2,3 disubstituted indoles is observed in the reaction of alkyl methyl ketones, and the products readily sublime directly from the ionic liquid. PMID- 14737529 TI - Facile alkane functionalization in copper-[2.1.1]-(2,6)-pyridinophane-PhINTs systems. AB - Mild catalytic dehydrogenation of cycloalkanes (cyclo-C(5)H(10), cyclo-C(6)H(12), cyclo-C(8)H(16)) and aziridination of resulting olefins is reported with PhINTs and copper-[2.1.1]-(2,6)-pyridinophane (L) complexes LCuX(n)(n= 1, 2; X = Cl, OTf)"activated" with NaBAr(F)(4) in dichloromethane solution. PMID- 14737528 TI - Chloride complexation by heptapeptides: influence of C- and N-terminal sidechains and counterion. AB - The channel-forming diglycolylated heptapeptides containing the amino acid sequence Gly-Gly-Gly-Pro-Gly-Gly-Gly have been found to complex chloride in CDCl(3). The strength of the interaction depends on the terminal alkyl groups and on chloride's countercation. PMID- 14737530 TI - Single-step in situ synthesis of double bond-grafted yttrium-hydroxide nanotube core-shell structures. AB - Novel MMA-Y(OH)(3) nanotube core-shell structures have been successfully prepared with double bonds successfully grafted on the surface through a single-step in situ hydrothermal method. PMID- 14737531 TI - Titanacyclopentene complexes and their application as 1,4-dicarbanion equivalents. AB - The treatment of Ti(OiPr)(4) with 3-butenylmagnesium chloride generates titanacyclopentene complexes which effectively add to carbonyl compounds and nitriles to afford the 1,4-coupling products with high Z:E selectivities. PMID- 14737532 TI - Intramolecular S(N)2' cyclization of an alkyllithium species onto a methoxy allyl ether is syn selective. AB - The preference for syn- or anti-addition of an intramolecular S(N)2[prime or minute] cyclization of an alkyllithium species onto a methoxy allyl ether has been proven unequivocally to take place by a syn S(N)2[prime or minute] mechanism. PMID- 14737533 TI - Selective deintercalation of apex over face-shared oxide ions in the topotactic reduction of Sr7Mn4O15 to Sr7Mn4O12. AB - Sodium hydride selectively deintercalates the apex rather than face-shared oxide ions within the structure of Sr(7)Mn(4)O(15) leading to the formation of the structurally related reduced phase Sr(7)Mn(4)O(12). PMID- 14737534 TI - An outer-sphere ligand for uranyl carbonate. AB - A novel supramolecular host for the uranyl carbonate complex has been designed and synthesized. The modified cyclodextrin host binds uranyl carbonate in water with a stability of 253 M(-1). PMID- 14737535 TI - Microwave-assisted free radical chemistry using the persistent radical effect. AB - Environmentally benign radical carboaminoxylations of various nonactivated olefins and difficult radical cyclization reactions are performed in good to excellent yields and with short reaction times under microwave irradiation. PMID- 14737536 TI - Selective host-guest interaction of single-walled carbon nanotubes with functionalised fullerenes. AB - Exohedrally functionalised fullerenes have been inserted in single-walled carbon nanotubes (SWNTs) with the aid of supercritical carbon dioxide to form peapods; C(61)(COOEt)(2) are encapsulated in SWNTs in high yield, whereas C(61)(COOH)(2) aggregate via hydrogen bonding to form a supramolecular complex, which sterically hinders encapsulation and causes it to adhere to the exterior surface of the SWNTs. PMID- 14737537 TI - Preparation of tripyrrane analogues from resorcinol and 2-methylresorcinol for applications in the synthesis of new benziporphyrin systems. AB - Acid catalyzed condensation of resorcinol or 2-methylresorcinol with 2 equiv. of an acetoxymethylpyrrole gave bis(pyrrolylmethyl)benzene derivatives in moderate yields; these afforded a series of novel aromatic benziporphyrins using the MacDonald "3 + 1" methodology. PMID- 14737538 TI - Synthesis and Bergman cyclization of a beta-extended porphyrenediyne. AB - Condensation of a porphyrin-2,3-dione with a 1,2-diaminoarenediyne affords a [small beta]-extended porphyrinic-enediyne: upon thermal Bergman cyclization the quinoxaline spacer positioned between the macrocycle and the enediyne prevents tandem radical cyclization to a picenoporphyrin. PMID- 14737539 TI - A fluorescent analogue of UDP-N-acetylglucosamine: application for FRET assay of peptidoglycan translocase II (MurG). AB - A direct continuous fluorescence assay for translocase II MurG based on fluorescence resonance energy transfer (FRET) has been developed using a 6 substituted fluorescent analogue of UDP-N-acetylglucosamine. PMID- 14737540 TI - A concise stereocontrolled formal total synthesis of (+/-)-podophyllotoxin using sulfoxide chemistry. AB - A short stereoselective formal total synthesis of (+/-)-podophyllotoxin has been carried out from a sulfoxide, using a one-pot tandem conjugate addition/aldol/electrophilic aromatic substitution reaction to form a tetralin, which was converted into picropodophyllin in two steps. PMID- 14737541 TI - A three dimensional porous metal-organic framework [Fe4L6.(DMF)3.(H2O)10] constructed from neutral discrete Fe4L6 pyramids [H2L = 1,3-benzodihydroxamix acid]. AB - A 3-D porous zeolite-like metal-organic framework surviving guest removal is assembled from a well-defined tetrahedral Fe(4)L(6) cavity by the cooperativity of hydrogen bonds and [small pi]-[small pi] stacking. PMID- 14737542 TI - A simple copper salt catalysed the coupling of imidazole with arylboronic acids in protic solvent. AB - In the presence of a catalytic amount of a simple copper salt, the coupling of imidazole with arylboronic acids was performed in methanol to give corresponding N-arylimidazoles in almost quantitative yields; this coupling reaction could also be performed in aqueous solutions to give N-arylimidazoles in excellent yields. PMID- 14737543 TI - 11-Aminoundecanoic acid: a versatile unit for the generation of low molecular weight gelators for water and organic solvents. AB - The use of 11-aminoundecanoic acid as a synthetic building-block allows the systematic preparation of (oligo)amide organogelators-including chiral ones-which display remarkable gelation properties in organic solvents and water. PMID- 14737544 TI - Synthesis and structural characterization of the first unsymmetrical diarylpalladium complex trans-Pd(C6F5)(2,4,6-C6F3H2)(PEt3)2, derived from transmetallation between 2,4,6-trifluorophenylboronic acid and trans Pd(C6F5)I(PEt3)2. AB - The reactions of 2,4,6-trifluorophenylboronic acid with aryl(iodo)palladium(ii) complexes, trans-Pd(C(6)F(5))I(PR(3))(2)(PR(3)= PEt(3), PMe(2)Ph, PMePh(2)) in the presence of Ag(2)O afforded trans-Pd(C(6)F(5))(2,4,6-C(6)F(3)H(2))(PR(3))(2) which are stabilized by fluorine atoms in the ortho positions. PMID- 14737545 TI - Intramolecular charge separation in a hydrogen bonded tyrosine-ruthenium(II) naphthalene diimide triad. AB - Long-lived charge-separated states in the ns to [micro sign]s range were observed upon laser flash excitation of a donor-chromophore-acceptor triad based on tris(bipyridine) ruthenium(ii) as photo-sensitizer, naphthalene diimide as acceptor, and a hydrogen bonded phenol as donor. PMID- 14737546 TI - Reactivation of aged model Pd/Ce0.68Zr0.32O2 three-way catalyst by high temperature oxidising treatment. AB - A deactivated (aged under redox-cycled model TWC feed-stream) Pd/Ce(0.68)Zr(0.32)O(2) catalyst is remarkably reactivated when subjected to a high temperature oxidising treatment whereas this effect is only marginal for Pd/Al(2)O(3), which indicates the key role of such treatment in restoring the Pd Ce(0.68)Zr(0.32)O(2) interactions leading to highly active catalysts. PMID- 14737547 TI - Cn microspheres as surrogate membranes in glycosidase-catalysed hydrolysis of glycolipids. AB - Glycosidase catalysed hydrolysis of glycolipids non-covalently attached to C(n) microspheres proceeds to completion for appropriate glycolipid-microsphere combinations in contrast with hydrolysis of covalently immobilised analogues which in all cases studied stops significantly short of complete hydrolysis. PMID- 14737548 TI - A chiroptical molecular switch with perfect stereocontrol. AB - A modified version of the first generation unidirectional molecular motor showed >99% stereoselectivity in photo-induced isomerizations in both directions, thus functioning as a perfect chiroptical molecular switch. PMID- 14737549 TI - Preparation of a novel diphosphine-palladium macrocyclic complex possessing a molecular recognition site. Oxidative addition studies. AB - A disphosphine-palladium(0) complex capable of recognising barbiturates has been prepared. Oxidative addition studies with a barbitiurate:aryl iodide conjugate provided new Pd(ii) complexes where the positioning of the Pd-bound aryl group is controlled by the molecular recognition event. PMID- 14737550 TI - Elimination of sulfur from aromatic heterocycles by a water-soluble arene ruthenium cluster: synthesis and molecular structure of [H2S2Ru4(C6H6)4]Cl2. AB - C-S bond cleavage in thiophene, benzothiophene and dibenzothiophene is achieved under biphasic conditions by the water-soluble cluster cation [H(4)Ru(4)(C(6)H(6))(4)](2+) which is converted into the disulfido cluster [H(2)S(2)Ru(4)(C(6)H(6))(4)](2+). PMID- 14737551 TI - Stepwise synthesis of siloxane chains. AB - Siloxane chains of designated lengths can be synthesized with high yields by reacting tris(tert-butoxy)silanol alternately with dichlorosilane and silanediol. PMID- 14737552 TI - Small molecule hydrogels based on a class of antiinflammatory agents. AB - Here we report small molecule hydrogels formed by the combination of two N (fluorenyl-methoxycarbonyl) amino acids, which belong to a novel class of antiinflammatory agents. PMID- 14737553 TI - Purification by HPLC and the UV/Vis absorption spectra of the nitrogen-containing incar-fullerenes iNC60, and iNC70. AB - We report the purification of the nitrogen-containing incar-fullerenes iNC(60) and iNC(70), and their characterisation by UV-Vis absorption spectroscopy. PMID- 14737554 TI - Unprecedented metal template effect on the coupling of dithiafulvene moieties. AB - Coordination of two dithiafulvenyldiphenylphosphines on a Mo(CO)(4) fragment allows a carbon-carbon bond formation upon oxidation, leading to a novel type of metallacycle substituted by a redox active vinylogous tetrathiafulvalene. PMID- 14737555 TI - Nuclear inelastic scattering spectroscopy of iron-sulfur cubane compounds. AB - The potential of NIS spectroscopy to study the iron-sulfur clusters in metalloproteins is illustrated using model compounds. The origin of the intense low energy transfer bands is discussed. PMID- 14737556 TI - An efficient chemical fixation of nitric oxide: convenient and practical synthesis of 1,2,3-oxadiazole 3-oxides. AB - Nitric oxide reacts efficiently with alkynyllithium at low temperature producing 1,2,3-oxadiazole 3-oxides in good yields. PMID- 14737557 TI - The first ligand-modulated oxidative Heck vinylation. Efficient catalysis with molecular oxygen as palladium(0) oxidant. AB - The discovery of the first ligand-supported Pd(ii) catalysed oxidative Heck reaction with molecular oxygen as reoxidant and the scope with diverse arylboronic acids and olefins using only 1-2% of catalyst are reported. PMID- 14737558 TI - Novel organic-inorganic composite coordination polymers generated from new multidentate Schiff-base ligand and Ag(I) salts. AB - Two novel organic-inorganic hybrid coordination polymers were synthesized based on a new multidentate Schiff-base ligand and Ag(i) salts PMID- 14737559 TI - A novel route to polyethylene-polystyrene blends through the fragmentation of porous polystyrene beads supported metallocene in ethylene polymerization. AB - Polyethylene-polystyrene blends were synthesized by in situ ethylene polymerization with polystyrene porous beads supported metallocene; the influence of fragmenting support beads on the morphology and the mechanical performance of the blends was investigated. PMID- 14737560 TI - A novel fluorescent indicator for Ba2+ in aqueous micellar solutions. AB - The highly selective and sensitive fluorescence detection of Ba(2+) among alkali metal and alkaline earth metal cations was successfully achieved in aqueous media by the combination of a novel monoazacryptand type of fluorophore and micelles of Triton X-100. PMID- 14737561 TI - Determination of the Lewis acidity of ionic liquids by means of an IR spectroscopic probe. AB - Pyridine and ethanenitrile can be used as molecular probes to measure the Lewis acidities of ionic liquids by monitoring the shift of IR absorption bands near 1450 cm(-1) for pyridine and in the range 2250-2340 cm(-1) for ethanenitrile. PMID- 14737562 TI - Fast racemisation and slow epimerisation of laterally lithiated amides: stereochemical evidence for the mechanism of inversion of amide-substituted benzyllithiums. AB - Tertiary 1-naphthamides racemise much more slowly than their laterally lithiated derivatives, and the relative rates of racemisation and epimerisation of these derivatives indicate that the lithium-bearing stereogenic centre inverts via a "conducted tour" mechanism, in which the lithium cation is delivered from one face to the other by coordination to the rotating amide group. PMID- 14737563 TI - Polymers of intrinsic microporosity (PIMs): robust, solution-processable, organic nanoporous materials. AB - Microporous materials can be derived directly from soluble polymers whose randomly contorted shapes prevent an efficient packing of the macromolecules in the solid state. PMID- 14737564 TI - Dichloro(dodeca-2,6,10-triene-1,12-diyl)ruthenium(IV): a highly efficient catalyst for the isomerization of allylic alcohols into carbonyl compounds in organic and aqueous media. AB - The catalytic activity of the bis(allyl)-ruthenium(iv) complex [Ru([small eta](3):[small eta](2):[small eta](3)-C(12)H(18))Cl(2)] in the transposition of allylic alcohols into carbonyl compounds, both in THF and H(2)O as solvent, is reported. PMID- 14737565 TI - Polyferrocenes: metallopolymers with tunable and high refractive indices. AB - The refractive index, molar refraction and Abbe number of polyferrocene derivatives are reported and the values indicate that these materials are very promising for a range of photonics applications. PMID- 14737566 TI - The intramolecular Baylis-Hillman reaction: easy preparation of versatile substrates, facile reactions, and synthetic applications. AB - We have developed a general and highly efficient method for the preparation of diverse [small omega]-formyl-[small alpha],[small beta]-unsaturated carbonyl compounds and optimized the conditions for the intramolecular Baylis-Hillman reactions of these compounds to provide various biologically important polycyclic compounds. PMID- 14737567 TI - Synthesis and ring opening reactions of a 2-silabicyclo[2.1.0]pentane. AB - Methyl 2-silabicyclo[2.1.0]pentane-1-carboxylate, obtained by a photochemical intramolecular cyclopropanation reaction of an [small alpha]-allylsilyl-[small alpha]-diazoacetate, undergoes ring opening reactions under different conditions leading to methyl 2-[diisopropyl(methoxy)silylmethyl]cyclopropane-1-carboxylate, a 1-sila-4-cyclopentene-2-carboxylate or an allyl(methoxysilyl)ketene. PMID- 14737568 TI - Synthesis and characterization of organic-inorganic hybrid mesoporous silica materials with new templates. AB - 1-Hexadecane-3-methylimidazolium bromide and 1-hexadecane-2,3-dimethylimidazolium bromide were used as new templates for the syntheses of periodic mesoporous organosilica (PMO) materials; using these new templates, ethane-bridged PMO materials were successfully synthesized and characterized under basic conditions. PMID- 14737569 TI - Photochemical oxidative addition of B-H bonds at ruthenium and rhodium. AB - Metal phosphine hydrides of type RuP(4)H(2) and RhP(3)H(3) react photochemically with HB(pin)(pin = pinacolate) to form metal boryl hydride complexes via 16 electron intermediates generated by H(2) loss; the second order rate constants for reaction of the intermediates with HB(pin) are even larger than those for reaction with Et(3)SiH. PMID- 14737570 TI - New efficient aerobic oxidation of some alcohols with dioxygen catalysed by N hydroxyphtalimide with vanadium co-catalysts. AB - New efficient vanadium co-catalysts have been developed for the oxidation of some alcohols with O(2) catalysed by N-hydroxyphthalimide (NHPI). Various alcohols (primary and secondary) were selectively oxidized by O(2) under mild conditions in the presence of a catalytic amount of NHPI as a radical-producing agent combined with small amounts of vanadium complexes with or without the addition of a simple salt (e.g. LiCl) or base (e.g. pyridine). PMID- 14737571 TI - The different role of Cu++ and Zn++ ions in affecting the interaction of prion peptide PrP106-126 with model membranes. AB - Differential scanning calorimetric (DSC) experiments have shown that the ability of PrP106-126 to perturb 1,3-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) model membranes is differently affected by Cu(++) and Zn(++) ions. PMID- 14737572 TI - On-chip electrochemical measurement of beta-galactosidase expression using a microbial chip. AB - [small beta]-Galactosidase expression in a small number of Escherichia coli cells has been measured in real time with an electrochemical sensor chip. E. coli cells were embedded using collagen gel within a micropore which was microfabricated onto a chip. The activity of the expressed [small beta]-galactosidase was determined using p-aminophenyl [small beta]-d-galactopyranoside (PAPG) as a substrate. PMID- 14737573 TI - Indium(I) trifluoromethanesulfonate and other soluble salts for univalent indium chemistry. AB - Herein we report the synthesis, structure and preliminary reactivity studies of a series of unusually soluble indium(i) salts that are improved alternatives to indium(i) halide reagents. PMID- 14737574 TI - Carbon nanotube screen-printed electrochemical sensors. AB - The fabrication, and evaluation of carbon-nanotube (CNT)-derived screen-printed (SP) electrochemical sensors based on a CNT ink are reported. The fabricated CNT strips combine the attractive advantages of CNT materials and disposable screen printed electrodes. Such thick-film CNT sensors have a well-defined appearance, are mechanically stable, and exhibit high electrochemical reactivity. PMID- 14737575 TI - Frequency-dependent electrical detection of protein binding events. AB - Frequency-dependent electrochemical impedance spectroscopy has been used to characterize the changes in electrical response that accompany specific binding of a protein to its substrate, using the biotin-avidin system as a model. Our results show that avidin, at concentrations in the nanomolar range, can be detected electrically in a completely label-free manner under conditions of zero average current flow and without the use of any auxiliary redox agents. Impedance measurements performed on biotin-modified surfaces of gold, glassy carbon, and silicon were obtained over a wide frequency range, from 5 mHz to 1 MHz. On each biotin-modified surface, binding of avidin is most easily detected at low frequencies, <1 Hz. Electrical circuit modeling of the interface was used to relate the frequency-dependent electrical response to the physical structure of the interface before and after avidin binding. Electrical measurements were correlated with measurements of protein binding using fluorescently labeled avidin. PMID- 14737577 TI - Analysis of erythromycin and tylosin in bovine muscle using disposable screen printed electrodes. AB - A disposable electrochemical enzyme-linked immunosorbent assay (ELISA) for the detection of two macrolides (erythromycin and tylosin) in bovine muscle was developed using a screen printed electrode (SPE) system as a differential pulse voltammetry (DPV) transducer with mouse anti-erythromycin (and anti-tylosin) monoclonal antibodies (MAb) serving as molecular recognition elements. The immunochemical system makes use of the competition assay principle, and employs an erythromycin (or tylosin)-BSA conjugate as coating molecule. After competition between free and coated analyte for the antibodies, the activity of the alkaline phosphatase labelled antiglobulins was measured electrochemically using 1 naphthylphosphate as substrate. Using standard solutions of erythromycin and tylosin, the detection limit of the assay was 0.2 ng mL(-1) determined to be for erythromycin and 2.0 ng mL(-1) for tylosin, while the sensitivity (25% inhibition concentration) was 1.0 ng mL(-1) for erythromycin and 3.0 ng mL(-1) for tylosin. The suitability of the assay for quantification of erythromycin and tylosin in bovine muscle was also studied. Spiked and real samples were analysed using the immunosensor system developed here. The ELISA showed precision values (relative standard deviation, RSD%) ranging from 4 to 9% for erythromycin and from 8 to 15% for tylosin; the accuracy (relative error, RE%) ranged from -11 to 6% and from -4 to 12% for erythromycin and tylosin, respectively. Results obtained on real samples were confirmed by micro-liquid chromatography coupled on line with tandem mass spectrometry (micro-LC-MS-MS), using an atmospheric pressure ionisation (API) source and an ionspray (IS) interface. The latter provides unequivocal identification and quantification of the analytes at the level of interest. PMID- 14737578 TI - Proton "off-on" behaviour of methylpiperazinyl derivative of naphthalimide: a pH sensor based on fluorescence enhancement. AB - In a search for new type pH sensing fluorophores, the possibility of using the proton "off-on" switch behaviour of naphthalimide derivatives for optical pH sensor preparation has been explored. A new compound, N-allyl-4-(4[prime or minute]-methyl-piperazinyl)-1,8-naphthalimide (AMPN), was synthesized. The enhancement of fluorescence of AMPN with the increase of hydrogen ion concentration is based on arresting photo-induced electron transfer to the naphthalimide fluorophore from aliphatic amine group after its protonation. The Stokes Shift of the proposed type of pH sensing fluorophore is significantly larger than that of the fluorescein counterparts. To avoid the leakage of the fluorophore, AMPN was photo-copolymerized with 2-hydroxyethyl methacrylate and acrylamide on the glass surface. The fluorescence intensity of membrane contacted with a pH 3.50 buffer is 4.7 times of that for pH 12.00 buffer solutions. The proposed pH sensor is not susceptible to ionic strength and shows good selectivity, repeatability and short response time. The membrane shows a good stability with a lifetime over two months. The sensor can be used for the determination of pH in the range of pH 4.5-9.0, without interference of most commonly co-existing inorganic ions and some organic species. The sensor has been applied to the analysis of urine samples. PMID- 14737581 TI - Liquid chromatography and tandem mass spectrometry: a powerful approach for the sensitive and rapid multiclass determination of pesticides and transformation products in water. AB - In this paper we will show the results of our research on the direct simultaneous determination of multi-class pesticides and transformation products with different polarities and acid-base properties by applying an on-line trace enrichment coupled to the chromatographic system supplied with electrospray interface (SPE-LC-MS/MS method). The specific chromatographic separation allows the correct determination of almost fifty compounds (37 pesticides and 10 transformation products) using very low sample volume and very little sample handling. Recoveries between 70-120% were obtained for all compounds in drinking and groundwater, meanwhile in surface water 44 compounds were correctly quantified. Relative standard deviations lower than 15% were obtained for all compounds. Even at the lowest concentration level tested (25 ng L(-1)) 40 compounds presented satisfactory recoveries and repeatability. The use of methanol as organic modifier and the increase of injection volume are also studied. The applicability of the developed method to a monitoring programme is demonstrated by applying it to the analysis of hundreds of samples. PMID- 14737579 TI - Integration of combined heteroduplex/restriction fragment length polymorphism analysis on an electrophoresis microchip for the detection of hereditary haemochromatosis. AB - This work describes an integrated method of enzymatic digestion, heteroduplex analysis (HA) and electrophoretic sizing on a microfluidic chip. HA techniques based on microchip electrophoresis are capable of the high sensitivity detection of subtle mutations such as single nucleotide polymorphisms (SNPs) but are not readily able to detect homozygous mutant genotypes. Such homozygous conditions are commonly encountered with the gene implicated in hereditary haemochromatosis, HFE. We employed the restriction fragment length polymorphism (RFLP) method of mutation detection to complement the HA method in a rapid novel on-chip procedure that separated digested PCR fragments to reliably determine the presence or absence of the most important mutations associated with haemochromatosis. This method was able to distinguish the homozygous mutant, heterozygous and homozygous wildtype genotypes. The mutations investigated here (C282Y, H63D and S65C) are often the mutation targets used in the genetic testing for haemochromatosis. This method provides the extremely specific digestion methods needed for the analysis of the known and relatively common mutations that have a significant probability of occurring in a homozygous form. However, the high sensitivity of the HA method is useful in detecting other mutations of lesser likelihood which, by virtue of their rarity, are likely to be present only in a heterozygous form. Although the conventional methods of analysing these mutations require as much as a day to perform, this microchip method, even without robotics or multiplexed operation, can be performed in about 10 min per sample. PMID- 14737582 TI - Sensitive and selective analysis of coenzyme Q10 in human serum by negative APCI LC-MS. AB - Two new sensitive and selective LC-MS methods have been developed for the quantification of the total coenzyme Q(10) concentration in human blood serum. The sensitivity of the methods is based on the very efficient formation of the radical anions of CoQ(10)[M(-)[radical dot]] by negative atmospheric pressure ionisation, APCI(-). The mass detection of the [M(-)[radical dot]] ions, m/z= 862.6, was performed either in selective ion monitoring (SIM) or in MS(2) mode (m/z= 862.6 [rightward arrow]m/z= 847.6) using an LCQ-deca ion-trap mass spectrometer. Two standard serum samples with medium (0.73 [micro sign]g ml(-1)) and high (1.96 [micro sign]g ml(-1)) total CoQ(10) concentrations were analysed by LC-APCI(-)-SIM and LC-APCI(-)-MS(2) and the results compared with a HPLC literature procedure with electrochemical detection (ECD). Both the LC-MS methods were shown to be more selective and with comparable or better sensitivity than the HPLC-ECD method. The LC-MS-SIM and LC-MS(2) chromatograms of the medium concentration sample showed CoQ(10) signal to noise ratios of 25 and 625, respectively. In addition, a simple and fast serum pre-treatment procedure was developed, in which the serum CoQ(10)H(2) content was quantitatively oxidised quantitatively to CoQ(10) in less than 15 min by 1,4-benzoquinone. PMID- 14737583 TI - Binding patterns of vanadium to transferrin in healthy human serum studied with HPLC/high resolution ICP-MS. AB - Vanadium (V) is an essential metal for mammals. It has different valence states. In blood, V is bound to transferrin (Tf), a glycoprotein that has two metal binding sites (C-lobe site and N-lobe site). In the present study, the binding patterns of V to serum Tf were analyzed by combined on-line HPLC and high resolution ICP-MS (HPLC/HR-ICP-MS). The levels of (51)V, (56)Fe and (32)S, which are interfered with polyatomic ions such as (35)Cl(16)O(+), (38)Ar(13)C(+) and (37)Cl(14)N(+), (40)Ar(16)O(+) and (40)Ca(16)O(+), and (16)O(2)(+), respectively, when using quadrupole ICP-MS, could be monitored simultaneously by HR-ICP-MS at a resolution of m/[capital Delta]m= 4000. Sample (a 1 ml portion of serum from a healthy person or 2 mg of human serum Tf (hTf)) was directly subjected to HPLC equipped with an anion-exchange column. V in human serum without any in vitro V spike was detected as V(C)-Tf (V bound to C-lobe site of Tf) and metal(2)-Tf. Since V(iii) was most favorable in terms of the binding to hTf in the presence of bicarbonate and V bound to the C-lobe site of hTf was detected only in the case of V(iii) among the three valence states of V, it was suggested that a part, at least, of V in the V(C)-Tf in healthy human serum may be present as V(iii), in addition to the generally accepted V(iv). PMID- 14737584 TI - On-site monitoring of biogenic emissions from Eucalyptus dunnii leaves using membrane extraction with sorbent interface combined with a portable gas chromatograph system. AB - Membrane extraction with sorbent interface, combined with a portable gas chromatograph system (MESI-Portable GC) for continuous on-line monitoring of biogenic volatile organic compounds (BVOCs) emissions (from leaves of Eucalytus dunnii in a greenhouse), is presented herein. A sampling chamber was designed to facilitate the extraction and identification of the BVOCs emitted by the Eucalytus dunnii leaves. Preliminary experiments, including; enrichment times, microtrap temperatures, stripping gas flow rates, and desorption temperatures were investigated to optimize experimental parameters. The main components of BVOCs released by the Eucalytus dunnii leaves were identified by comparing the retention times of peaks with those of authentic standard solutions. They were then confirmed with solid phase microextraction coupled with gas chromatography and mass spectrometry (SPME-GC-MS). BVOC emission profiles of [small alpha] pinene, eucalyptol, and [gamma]-terpinene emitted by intact and damaged Eucalytus dunnii leaves were obtained. The findings suggest that the MESI-Portable GC system is a simple and useful tool for field monitoring changes in plant emissions as a function of time. PMID- 14737585 TI - Immunoassay using probe-labelling immunogold nanoparticles with silver staining enhancement via surface-enhanced Raman scattering. AB - This paper reports a novel immunoassay based on surface-enhanced Raman scattering (SERS) and immunogold labelling with silver staining enhancement. Immunoreactions between immunogold colloids modified by a Raman-active probe molecule (e.g., 4 mercaptobenzoic acid) and antigens, which were captured by antibody-assembled chips such as silicon or quartz, were detected via SERS signals of Raman-active probe molecule. All the self-assembled steps were subjected to the measurements of ultraviolet-visible (UV-vis) spectra to monitor the formation of a sandwich structure onto a substrate. The immunoassay was performed by a sandwich structure consisting of three layers. The first layer was composed of immobilized antibody molecules of mouse polyclonal antibody against Hepatitis B virus surface antigen (PAb) on a silicon or quartz substrate. The second layer was the complementary Hepatitis B virus surface antigen (Antigen) molecules captured by PAb on the substrate. The third layer was composed of the probe-labelling immunogold nanoparticles, which were modified by mouse monoclonal antibody against Hepatitis B virus surface antigen (MAb) and 4-mercaptobenzoic acid (MBA) as the Raman active probe on the surface of gold colloids. After silver staining enhancement, the antigen is identified by a SERS spectrum of MBA. A working curve of the intensity of a SERS signal at 1585 cm(-1) due to the [small nu](8a) aromatic ring vibration of MBA versus the concentration of analyte (Antigen) was obtained and the non-optimized detection limit for the Hepatitis B virus surface antigen was found to be as low as 0.5 [micro sign]g mL(-1). PMID- 14737587 TI - Application of wavelet transforms and an approximate deconvolution method for the resolution of noisy overlapped peaks in DNA capillary electrophoresis. AB - A new procedure for resolving noisy overlapped peaks in DNA separations by capillary electrophoresis (CE) is developed. The procedure combines both a wavelet-based denoising method that effectively denoises the signal and a novel approximate deconvolution technique that resolves the fragment peaks and improves the ability to separate highly overlapped peaks early in the electrophoresis process. Different kinds of overlapped peaks with and without noise simulated by computer as well as some DNA experimental electropherograms were submitted to the new procedure. A second order differential operator with variable coefficients is applied to the entire electrophoresis signal at any given time and approximate deconvolutions of the individual Gaussian peaks are performed. The operator incorporates the effect of the superposition and gives exact annihilation in the neighborhood of each peak. Overlapped peaks with a resolution higher than 0.46 can be resolved directly. Also, the method can determine the peak components of signals with a signal to noise ratio higher than 1.4 PMID- 14737589 TI - Immunoquantitative analysis for berberine and its related compounds using monoclonal antibodies in herbal medicines. AB - The monoclonal antibody (MAb) against berberine, a bioactive constituent of Coptis japonica M., Phellodendron amurense R. and Hydrastis canadensis L., was produced and characterized. As immunogen, the derivative of berberine, 9-O carboxymethyl berberrubine was synthesized and conjugated to carrier protein, bovine serum albumin (BSA). In order to confirm its immunogenicity, the ratio of hapten in berberine-BSA conjugate was determined by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS). After immunization, hybridomas secreting MAbs against berberine were produced by fusing splenocytes with mouse myeloma cell line, P3-X63-Ag8-653. After the screening, anti-berberine MAb 1D5-3B-7 was obtained. Subsequently, a quantitative ELISA system for berberine and its related compounds using the MAb was established and evaluated comparing with HPLC method. The ELISA method described in this study can be available as an analytical tool for quality control and standardization of medicinal plants and its prescriptions containing berberine and its related compounds. PMID- 14737590 TI - Comment on "Soxhlet extraction of acrylamide from potato chips" by J. R. Pedersen and J. O. Olsson, Analyst, 2003, 128, 332. PMID- 14737591 TI - Comment on "Soxhlet extraction of acrylamide from potato chips" by J. R. Pedersen and J. O. Olsson, Analyst, 2003, 128, 332. PMID- 14737592 TI - Comment on "Soxhlet extraction of acrylamide from potato chips" by J. R. Pedersen and J. O. Olsson, Analyst, 2003, 128, 332. PMID- 14737634 TI - Diastereotopic group selective intramolecular cycloadditions of sulfenic acids to 1,4-dienes. AB - The ratio of diastereomeric cis-fused perhydrobenzothiophene S-oxides formed via the intramolecular addition of a sulfenic acid to a 1,4-diene is controlled by the nature of the protecting group on a chiral alcohol in the connecting chain. PMID- 14737635 TI - The effect of pressure on microwave-enhanced Diels-Alder reactions. A case study. AB - It is demonstrated that microwave-assisted Diels-Alder reactions of substituted 2(1H)-pyrazinones with ethylene are significantly more effective utilizing pre pressurized (up to 10 bar) reaction vessels. PMID- 14737636 TI - The influence of chiral auxiliaries and catalysts on the selectivity of intramolecular conjugate additions of pyrrole to N-tethered Michael acceptors. AB - A series of pyrroles incorporating N-tethered acrylates and related groups has been prepared and examined for their capacity to undergo intramolecular Michael addition reactions to form, in a diastereo- or enantio-selective fashion, the corresponding 8-substituted tetrahydroindolizidine or homologues thereof. PMID- 14737637 TI - Microwave assisted Leimgruber-Batcho reaction for the preparation of indoles, azaindoles and pyrroylquinolines. AB - The development of enhanced conditions for Lewis acid catalysed Leimgruber-Batcho indole synthesis using microwave acceleration is described. This approach has permitted the preparation of a variety of heteroaromatic enamine intermediates in good yield and high purities. Subsequent catalytic hydrogenation reactions, under various conditions including the use of a solid-phase encapsulated catalyst, furnish the corresponding indole derivatives in good yields. PMID- 14737638 TI - Reactive resin facilitated preparation of an enantiopure fluorobicycloketone. AB - A facile preparation of enantiopure ethyl (1S,5S,6S)-6-fluoro-2 oxobicyclo[3.1.0]hexane-6-carboxylate is described. The key feature of the synthesis involves copper-catalyzed enantioselective intramolecular cyclopropanation of a diazoketone to form endo-fluorocyclopropane in a single operation. Removal of a problematic chloroketone impurity using a reactive resin treatment enabled a high throughput enantiopurity upgrade by chiral HPLC. The development of a scalable synthesis of is presented, including details of the selection of catalyst and ligand optimization, incorporation of a reactive resin treatment and selection of chiral HPLC media and conditions. PMID- 14737639 TI - Pi-halogen dimer interactions and the inclusion chemistry of a new tetrahalo aryl host. AB - The preparation of 1,4,8,11-tetrabromo-5b[small alpha],6,12b[small alpha],13 tetrahydropentaleno[1,2-b:4,5-b[prime or minute]]diquinoline is described. This is a further member of the tetrahalo aryl host family, and forms crystalline lattice inclusion compounds with many guests. The X-ray structures of the allyl cyanide, 1,2,3-trichloropropane, chlorobenzene, toluene, benzene-water, methyl chloroform and carbon tetrachloride inclusion compounds are described, and compared with that of the solvent-free apohost. Although several different structural types are produced, the recently reported pi-halogen dimer (PHD) interaction plays an important role in all of these, except for that of pure (where the packing energy is the least favourable of the series). PMID- 14737640 TI - Synthesis and hybridization properties of l-oligodeoxynucleotide analogues fixed in a low anti glycosyl conformation. AB - We have synthesized l-type enantiomers (cU and cA) of nucleoside analogues, whose glycosyl bonds are fixed in a low anti conformation (ap glycosyl conformation, [small chi][approximate] 180[degree]), and incorporated them into oligonucleotides to evaluate the hybridization ability with natural DNA and RNA sequences. Although the incorporation of the modified nucleosides into oligonucleotides decreased the hybridization ability with unmodified complementary DNA sequences, the fully-substituted 12mers (cU(12) and cA(12)) still retained the hybridization ability with the complementary unmodified DNA 12mers, regardless of their unnatural l-chirality. In contrast, cU(12) and cA(12) showed different hybridization behavior with complementary unmodified RNA 12mers. cU(12) forms a more stable duplex with rA(12) than the corresponding natural 12mer (dT(12)), whereas cA(12) cannot hybridize with rU(12). Based on the model structure of cU(12)-rA(12), we discuss these experimental results. PMID- 14737641 TI - Formation of the heterocumulene anion SCCCN(-) by a cyano migration from the radical anion of 1,2-dicyanoethylenedithiolate. AB - The anionic heterocumulene SCCCN(-) was generated in the gas phase by collisional activation of the radical anion of 1,2-dicyanoethylenedithiolate. The mechanism of this reaction, as well as the structures of neutral and anionic products, was investigated by hybrid density functional theory (DFT) calculations. Dissociation to form SCCCN(-) and SCN is proposed to occur by a radical directed cyano migration reaction, with calculations suggesting this is the lowest energy fragmentation pathway available to the precursor anion. In contrast, the even electron protonated 1,2-dicyanoethylenedithiolate anion fragmented by loss of HCN. PMID- 14737642 TI - Ketene-acetylene [2 + 2] cycloadditions: cyclobutenone and/or oxete formation? AB - The [2 + 2] cycloaddition of monosubstituted acetylenes to ketene has been studied by ab initio(G2(MP2,SVP) and DFT (B3LYP/6-31Gd)) methods. The activation barrier decreases with increasing electron-donating ability of the acetylene substituent, and it can be roughly correlated with the energy of the acetylene HOMO. The addition to the C[double bond, length as m-dash]C bond of ketene (giving cyclobutenones) is preferred for the less electron-rich acetylenes, but for the most electron rich ones (X = NH(2) and NMe(2)) the addition to the C[double bond, length as m-dash]O bond (giving oxetes) becomes competitive, with activation barriers as low as ca. 45 (30) kJ mol(-1) for the two computational methods used. The cyclobutenones and oxetes can undergo ring opening to vinylketenes and acylallenes, respectively. Furthermore, the latter two compounds can interconvert by a 1,3-shift of the substituent X. The acylallenes become thermodynamically more stable than the vinylketenes for [small pi]-(lone pair) donating substituents X, and the 1,3-shift barrier also decreases, to ca. 130 kJ mol(-1) for X = NMe(2). In contrast, the 1,3-shifts of CH(3) and H have very high barriers. PMID- 14737643 TI - Ab initio and NMR studies on the effect of hydration on the chemical shift of hydroxy protons in carbohydrates using disaccharides and water/methanol/ethers as model systems. AB - Density functional theory (DFT) and Hartree-Fock (HF) quantum mechanical calculations have been performed on the disaccharides, [small beta]-l-Fucp (1[rightward arrow]4)-[small alpha]-d-Galp-OMe, [small beta]-l-Fucp-(1[rightward arrow]4)-[small alpha]-d-Glcp-OMe, and [small beta]-l-Fucp-(1[rightward arrow]3) [small alpha]-d-Glcp-OMe. The [capital Delta][small delta]-values (difference between the chemical shift in the disaccharide and the corresponding monosaccharide methyl glycoside) for the exchangeable hydroxy protons have been calculated and compared to experimental values previously measured by NMR spectroscopy for samples in aqueous solutions. The calculations performed on molecules in vacuum showed that hydroxy protons hydrogen bonded to the neighboring ring oxygens have large positive [capital Delta][small delta]-values, indicating that they are deshielded relative to those in the corresponding methyl glycoside. The NMR experiments showed instead that these hydroxy protons close to the neighboring ring oxygens were shielded. This discrepancy between calculated and experimental data was attributed to solvent effects, and this hypothesis has been confirmed in this work by monitoring the chemical shift of the hydroxy proton of methanol in water, ethers and water/ether solutions. Shielding of the hydroxy proton of methanol is observed for increased ether concentrations, whereas deshielding is observed for increased concentration of water. The shielding observed for hydroxy protons in disaccharides is a consequence of reduced hydration due to intermolecular hydrogen bonding or steric effects. In strongly hydrated systems such as carbohydrates, the hydration state of a hydroxy proton is the key factor determining the value of the chemical shift of its NMR signal, and the [capital Delta][small delta] will be a direct measure of the change in hydration state. PMID- 14737644 TI - A stereochemical anomaly: the cyclised (R)-AMPA analogue (R)-3-hydroxy-4,5,6,7 tetrahydroisoxazolo[5,4-c]pyridine-5-carboxylic acid [(R)-5-HPCA] resembles (S) AMPA at glutamate receptors. AB - (RS)-3-Hydroxy-4,5,6,7-tetrahydroisoxazolo[5,4-c]pyridine-5-carboxylic acid (5 HPCA)(), which is a conformationally constrained cyclised analogue of AMPA has previously been described as causing glutamate receptor mediated excitations of spontaneously firing cat spinal interneurons in a similar fashion to AMPA. We have now prepared the enantiomers of through chiral chromatographic resolution of (RS)-3-(carboxymethoxy)-4,5,6,7-tetrahydroisoxazolo[5,4-c]pyridine-5-carboxylic acid () followed by a stereoconservative hydrolysis resulting in the enantiomers of with high enantiomeric excess (% ee [greater-than-or-equal] 99). The absolute configurations indicated by an X-ray analysis of (-)- monohydrate were confirmed by comparing observed and ab initio calculated electronic circular dichroism spectra and by stereoconservative synthesis of (S)- from (S)-AMPA, the pharmacologically active form of AMPA. The pharmacological effects at native and cloned (GluR1-4) AMPA receptors were shown to reside exclusively with (R)-(+)-, in striking contrast to the usual stereoselectivity trend among AMPA receptor agonists. The reasons for this anomalous behaviour became clear upon docking both enantiomers of to the agonist binding site of GluR2. PMID- 14737646 TI - Synthesis and properties of bioactive 2- and 3-amino-8-methyl-8H-quino[4,3,2 kl]acridine and 8,13-dimethyl-8H-quino[4,3,2-kl]acridinium salts. AB - Cyclisation of 9-(benzotriazol-1-yl)acridine to the pentacycle 8H-quino[4,3,2 kl]acridine in a range of low-boiling solvents is mechanistically distinct from previously published photochemical (carbene) and thermolytic (radical) cyclisations. Fragmentation of the triazole ring of to a diazonium intermediate, and its subsequent heterolysis (-N(2)) and cyclisation is facilitated by solvation of intermediate zwitterionic species. Derivatives of 2- and 3 aminoquinoacridines methylated in the 8-position can be converted to 8,13 dimethylquino[4,3,2-kl]acridinium iodide salts with methyl iodide and were required for biological examination as potential telomerase inhibitors. The chloro group in 3-chloro-8-methyl-8H-quino[4,3,2-kl]acridine can be replaced efficiently by benzylamino, 4-morpholinyl and cyano substituents in palladium(0) mediated reactions. PMID- 14737645 TI - Facilitated phosphatidylserine flip-flop across vesicle and cell membranes using urea-derived synthetic translocases. AB - Tris(2-aminoethyl)amine derivatives with appended urea and sulfonamide groups are shown to facilitate the translocation of fluorescent phospholipid probes and endogenous phosphatidylserine across vesicle and erythrocyte cell membranes. The synthetic translocases appear to operate by binding to the phospholipid head groups and forming lipophilic supramolecular complexes which diffuse through the non-polar interior of the bilayer membrane. PMID- 14737647 TI - Synthesis and antiviral activity of monofluorinated cyclopropanoid nucleosides. AB - Diastereopure monofluorinated cyclopropanoid nucleosides were synthesized for biological studies. As key intermediates cis- and trans-(+/-)-[1-fluoro-2 (acetoxymethyl)cyclopropyl]methanol were prepared starting from diastereopure fluorinated cyclopropanecarboxylates. The latter were synthesized by copper(i) catalyzed cyclopropanation of [small alpha]-fluorostyrene with ethyl diazoacetate. After reduction and O-acetylation the diastereomeric (2-fluoro-2 phenylcyclopropyl)methyl acetates were obtained. Oxidative degradation using RuO(4) and reduction of the formed carboxyl group with borane gave the fluorinated alcohols, which were coupled with different nucleobases. After deprotection, the corresponding cyclopropanoid nucleosides of adenine, cytosine, guanine, thymine and uracil were obtained. Antiviral tests revealed for the cis configured guanosine a low, but specific activity against HSV-1 and HSV-2. In addition low affinities of the adenine derivatives to adenosine receptors were detected. PMID- 14737648 TI - Probing the stereochemistry of the active site of gamma-glutamyl transpeptidase using sulfur derivatives of l-glutamic acid. AB - Gamma-glutamyl transpeptidase (GGT) catalyses the transfer of a gamma-glutamyl moiety from a donor substrate to different acceptors, such as amino acids and water. GGT is known to display relatively low stereospecificity with respect to the alpha-stereocentre of its donor substrates. In this study we have studied its stereospecificity with respect to the stereocentre at the delta-position of different analogues of L-glutamic acid. Notably, L-methionine sulfoxide is well recognised whereas L-methionine sulfone and L-methionine sulfoximine are not. Furthermore, when the synthetic gamma-diastereoisomers of L-methionine sulfoxide were separated and tested, it was discovered that GGT shows remarkable stereospecificity at the gamma-position, binding the S(C)S(S) diastereoisomer with a K(i) of 3.5 mM, whereas the S(C)R(S) diastereoisomer is not recognised. Finally, using a sulfoxide as a new pharmacophore for GGT, we have synthesized and tested an analogue of glutathione to obtain a very promising competitive inhibitor with a K(i) of (53 +/- 3) microM. PMID- 14737649 TI - Synthesis of 1,3-diazepines and ring contraction to cyanopyrroles. AB - Several tetrazolo[1,5-a]pyridines/2-azidopyridines undergo photochemical nitrogen elimination and ring expansion to 1,3-diazacyclohepta-1,2,4,6-tetraenes, as well as ring cleavage to cyanovinylketenimines, in low temperature Ar matrices. 6,8 Dichlorotetrazolo[1,5-a]pyridine/2-azido-3,5-dichloropridine undergoes ready exchange of the chlorine in position 8 (3) with ROH/RONa. 8-Chloro-6 trifluoromethyltetrazolo[1,5-a]pyridine undergoes solvolysis of the CF(3) group to afford 8-chloro-6-methoxycarbonyltetrazolo[1,5-a]pyridine. Several tetrazolopyridines/2-azidopyridines afford 1H- or 5H-1,3-diazepines in good yields on photolysis in the presence of alcohols or amines. 5-Chlorotetrazolo[1,5 a]pyridines/2-azido-6-chloropyridines and undergo a rearrangement to 1H- and 3H-3 cyanopyrroles and, respectively. The mechanism of this rearrangement was investigated by (15)N-labelling and takes place via transient 1,3-diazepines. The structures of 6,8-dichloro-tetrazolo[1,5-a]pyridine, 6-chloro-8 ethoxytetrazolo[1,5-a]pyridine, dipyrrolylmethane, and 2-isopropoxy-4 dimethylamino-5H-1,3-diazepine were determined by X-ray crystallography. In the latter case, this represents the first reported X-ray crystal structure of a 5H 1,3-diazepine. PMID- 14737650 TI - Lewis acid-promoted cyclization of heteroatom-substituted enynes. AB - Lewis acid-promoted cyclizations of heteroatom-substituted enynes have been examined. The reaction of enynes and bearing silicon substituents on an alkyne afforded the halogenated five-membered gamma-lactones and gamma-lactams as the main products. The reaction of substrates and having 2-phosphonoacrylate instead of malonate also gave halogenated five-membered cyclic compounds and as the major products. The cyclized products are highly substituted and potentially useful for further synthetic transformations. PMID- 14737651 TI - Asymmetric synthesis of N-protected amino acids by the addition of organolithium carboxyl synthons to ROPHy/SOPHy-derived aldoximes and ketoximes. AB - A new asymmetric synthesis of alpha-amino acids is described in which the key step is the highly diastereoselective addition of organolithium carboxyl synthons (2-furyllithium, phenyllithium, vinyllithium) to (R)- and (S)-O-(1-phenylbutyl) oximes to give hydroxylamines, with vinyllithium being the most satisfactory nucleophilic reagent. Subsequent reductive cleavage of the N-O bond in hydroxylamines, followed by N-protection, and oxidative cleavage of the carboxyl precursor gave a range of N-protected amino acids and esters. The method was exemplified by the synthesis of a range of derivatives of non-proteinogenic amino acids such as 4-bromophenylalanine, tert-leucine, norvaline, cyclohexyl- and aryl glycines, 2-amino-8-oxodecanoic acid (Aoda) and alpha-methylvaline. PMID- 14737652 TI - A 'molecular switchboard'--covalent modifications to proteins and their impact on transcription. AB - Proteins undergo a remarkable variety of posttranslational modifications, with more than 200 distinct modifications identified to date. Increasing evidence suggests that many proteins bear multiple, distinct modifications, and the ability of one modification to antagonize or synergize the deposition of another can have significant biological consequences. Here, we illustrate the importance of posttranslational modifications within the context of transcriptional regulation, and we offer a perspective on the emerging role of combinatorial networks of modifications. Finally, we discuss the potential for chemical approaches to transform our understanding of the field. PMID- 14737653 TI - Ru complexes bearing bidentate carbenes: from innocent curiosity to uniquely effective catalysts for olefin metathesis. AB - The discovery and development of a new class of Ru-based catalysts for olefin metathesis is described. These catalysts, particularly those that do not bear a phosphine ligand, have been demonstrated to promote unique levels of reactivity in a variety of olefin metathesis reactions. The design and development of supported and chiral optically pure variants of this class of Ru catalysts for use in enantioselective metathesis are discussed as well. All catalysts are air stable, reusable, and can be employed with unpurified solvents. PMID- 14737654 TI - Rapid methylation of terminal acetylenes by the Stille coupling of methyl iodide with alkynyltributylstannanes: a general protocol potentially useful for the synthesis of short-lived 11CH3-labeled PET tracers with a 1-propynyl group. AB - The Pd(0)-mediated rapid coupling (trapping) reaction of methyl iodide with an excess amount of alkynyltributylstannane has been developed with the aim to incorporate a short-lived (11)C-labeled methyl group into biologically active organic compounds with a 1-propynyl structural unit. PMID- 14737655 TI - A novel and facile synthesis of dienals and substituted 2H-pyrans via the Vilsmeier reaction of alpha-oxo-ketenedithioacetals. AB - A novel and facile synthesis of dienals (3a, 3b) and substituted 2H-pyrans (4c, 4d) from a series of [small alpha]-oxo ketenedithioacetals containing a methyl group adjacent to the carbonyl group (1a-d)via the Vilsmeier reaction has been developed and a mechanism for the reactions has been proposed. PMID- 14737656 TI - Synthesis of an S-linked glycopeptide analog derived from human Tamm-Horsfall glycoprotein. AB - Direct base catalyzed S-glycosylation of a cysteine and a homocysteine containing peptide with O-acetyl protected bromides in DMF-water solution furnished two glycopeptide fragments. The two glycopeptide fragments were linked to the target glycopeptide with two S-glycosyl residues mimicking a part of Tamm-Horsfall glycoprotein. PMID- 14737657 TI - Diamagnetic and paramagnetic ring currents in expanded porphyrins. AB - Ipsocentric current density maps are computed at the coupled Hartree-Fock level in the 6-31G** basis set for the planar C(2v) B3LYP geometries of the expanded porphyrins, sapphyrin and orangarin. Both give clearly dominant global macrocyclic ring currents, but with opposite senses of circulation: in 22[small pi] sapphyrin, a diatropic current runs, with some bifurcation, around the conventional 22-centre delocalisation pathway; in 20[small pi] orangarin, a paratropic current runs around the inner 17-atom pathway. In agreement with the annulene analogy for these macrocycles, analysis of orbital contributions shows that in each case topology, energy and symmetry of the frontier orbitals together determine the macrocyclic ring current. In sapphryrin, 4-electron diamagnetism (aromaticity) arises from translationally allowed HOMO-LUMO excitations as in benzene itself; in orangarin, 2-electron paramagnetism (antiaromaticity) arises from rotationally allowed HOMO-LUMO excitations as in planarised cyclooctatetraene. The active orbitals invoked in the explanation of ring currents are those involved in the longstanding four-orbital model of porphyrin electronic spectra. PMID- 14737658 TI - Total synthesis of (-)-microcarpalide, a novel microfilament disrupting metabolite. AB - The stereoselective total synthesis of (-)-microcarpalide, a recently discovered 10-membered lactone of fungal origin displaying a remarkable disrupting action on actin microfilaments, was accomplished by using ring-closing metathesis (RCM) as the key step for the formation of the medium-sized ring. The diene ester required for the macrocyclization reaction was assembled via DCC-mediated esterification of two suitable partners, each bearing a terminal alkene group. The alcohol fragment was synthesized from n-bromohexane through a seven-step sequence entailing two consecutive stereoselective homologations of chiral boronic esters as strategic transformations for the sequential insertion of the two stereocentres with the final S absolute configuration, using (+)-pinanediol as the chiral director; final elaboration to the desired C(11) framework envisaged treatment with an allyl Grignard reagent and oxidative cleavage of the boronic scaffold. In contrast, the acidic fragment was prepared in ten steps from d tartaric acid, whose C(4) backbone was elongated to the required C(7) skeleton by means of two distinct Swern-Wittig oxidation-homologation sequences. PMID- 14737659 TI - Absolute stereochemistry of dihydrofuroangelicins bearing C-8 substituted double bonds: a combined chemical/exciton chirality protocol. AB - Coumarins are associated with a variety of pharmacological activities which have led to the synthesis of numerous derivatives. However, no general method for determination of the absolute configuration of chiral coumarins is known. This has now been achieved for a series of dihydrofuroangelicins bearing a variety of C-8 substituted double bonds, synthesized in the racemic form and resolved through enantioselective chromatography. A combined chemical/chiroptical protocol has been developed in which the C[double bond, length as m-dash]C double bonds are replaced with a styrenoid chromophore through either (i) cross metathesis, (ii) Heck reaction, or (iii) a combined method of cross metathesis and Heck reaction with about 1 mg sample under mild conditions. The coupling between the styrenoid and coumarin chromophores gives rise to clear-cut exciton coupled CD curves, suitable for assignments of absolute configurations. The solution conformation of the styrenoid derivatives is determined by NMR and DFT molecular modeling; the electronic structure of the 7-hydroxy coumarin chromophore is also clarified by semi-empirical and TDDFT methods. The conformation thus derived, in conjunction with quantitative DeVoe's coupled-oscillator CD calculation, establishes the absolute configurations of the coumarins. The theoretical study described herein justifies the straightforward approach of the current chemical/exciton chirality protocol to this type of dihydrofuroangelicins. PMID- 14737660 TI - New isocysteine building blocks and chemoselective peptide ligation. AB - Boc-, Fmoc- and Cbz-protected isocysteine building blocks were prepared by a concise three-step procedure starting from thiomalic acid. The use of Boc/Trt protected isocysteine provided convenient access to isocysteinyl peptides that allow the chemoselective ligation of unprotected peptide fragments in water. The pH-dependency of the isocysteine-mediated ligation was compared with that of cysteine-mediated native chemical ligation. PMID- 14737661 TI - Stabilisation of the transition state of phosphodiester bond cleavage within linear single-stranded oligoribonucleotides. AB - The effect of base sequence on the stability of the transition state (TS) of phosphodiester bond cleavage within linear single-stranded oligoribonucleotides has been studied in order to better understand why the reactivity of some phosphodiester bonds is enhanced compared to an unconstrained linkage. Molecular dynamics simulations of 3.0 ns were carried out for 14 oligonucleotides that contain in the place of the scissile phosphodiester bond a phosphorane structure mimicking the TS of the bond cleavage. The hydrolytic stability of the same oligonucleotides had previously been reported. Both the non-bridging oxyanions and the leaving 5[prime or minute]-oxygen of the pentacoordinated phosphorane moiety were observed to form hydrogen bonds with solvent water molecules in a similar way with all the compounds studied. In addition, water mediated hydrogen bonds between the phosphorane non-bridging oxyanions and the bases of the 3[prime or minute]-flanking sequence were detected with some of the compounds, but not with the most labile ones. Hence, it seems that the enhanced cleavage of some internucleosidic linkages does not result from the TS stabilisation by hydrogen bonding. With heterooligomers, the stacking of bases next to the cleavage site was observed to be enhanced on going from the initial state to the TS, whereas within uracil homooligomer, having initially negligible stacking, no change in the magnitude of stacking was seen. Accordingly, while strong stacking in the initial state is known to retard the phosphodiester bond cleavage, it may in the TS accelerate the reaction. Therefore, enhanced stacking on going from the initial state to transition state appears to be a factor that markedly contributes to the hydrolytic stability of phosphodiester bonds within oligonucleotides and may, at least partly, explain accelerated cleavage compared to fully unconstrained bonds, such as those in polyuridylic acid. PMID- 14737662 TI - Cyclic PNA-based compound directed against HIV-1 TAR RNA: modelling, liquid-phase synthesis and TAR binding. AB - A cyclic molecule including a hexameric PNA sequence has been designed and synthesized in order to target the TAR RNA loop of HIV-1 through the formation of a "kissing complex". For comparison, its linear analogue has also been investigated. The synthesis of the cyclic and linear PNA has been accomplished following a liquid-phase strategy using mixed PNA and fully N-protected (aminoethylglycinamide) fragments. The interactions of this cyclic PNA and its linear analogue with TAR RNA have been studied and the results indicate clearly that no interaction occurs between the cyclic antisense PNA and TAR RNA, whereas a tenuous interaction has been detected with its linear PNA analogue. PMID- 14737664 TI - Atom-efficient electrophilic aromatic nitration by dinitrogen pentoxide catalysed by zirconium(IV) 2,4-pentanedionate. AB - An atom-efficient, non-acidic, catalytic process is described for the nitration of electron deficient arenes such as o-nitrotoluene using a dinitrogen pentoxide zirconium(iv) 2,4-pentanedionate system in dichloromethane solvent. Kinetic studies showed the nitration process to be first-order with respect to the aromatic substrate and higher than first-order with respect to the catalyst. Addition of the catalyst at ca. 0.1-1 mol% compared with both N(2)O(5) and the organic substrate results in an increase in the first-order rate constant for nitration by a factor of approximately 5000 with a turnover number of at least 500. The orientation of the nitration products (2,4-/2,6-dinitrotoluenes) is consistent with attack of nitronium ion. The apparently high order of reaction with respect to the catalyst suggests a possible heterogeneous process. PMID- 14737663 TI - Oligodeoxynucleotides containing alpha-L-ribo configured LNA-type C-aryl nucleotides. AB - Synthesis of 2[prime or minute]-O,4[prime or minute]-C-methylene-[small alpha]-l ribofuranosyl derivatives containing phenyl and 1-pyrenyl aglycons, i.e., novel [small alpha]-l-ribo configured LNA-type C-aryl nucleosides, has been accomplished. Key synthetic steps included stereoselective Grignard reactions on tetrahydrofuran aldehyde, configurational inversion of the resulting alcohol into alcohol, and concomitant Mitsonobu cyclization furnishing the desired bicyclic furanosyl skeleton with a locked conformation. The phosphoramidite derivatives and were used for automated synthesis of 9-mer DNA and [small alpha]-L-LNA oligonucleotides containing the [small alpha]-L-LNA-type C-aryl monomers ([small alpha]L)Ph(L) and ([small alpha]L)Py(L) containing a phenyl and pyrenyl aglycon, respectively. Thermal denaturation studies showed universal base pairing behavior for the pyrenyl monomer ([small alpha]L)Py(L) when incorporated into a DNA or an [small alpha]-L-LNA oligonucleotide. PMID- 14737665 TI - The fluorine-containing pi-allylmetal complex. The transition metal-catalyzed allylic substitution reaction of fluorinated allyl mesylates with various carbon nucleophiles. AB - The allylic substitution reaction of [small alpha]-fluoroalkylated allyl mesylates with various carbon nucleophiles in the presence of transition metal catalyst (Pd and Mo) proceeded with high regioselectivity to give the corresponding [gamma]-fluoroalkylated products in excellent yields. PMID- 14737666 TI - Esterase-assisted accumulation of 3-carboxy-2,2,5,5-tetramethyl-1 pyrrolidinyloxyl into lymphocytes. AB - The intracellular detection of hydroxyl radical (HO*) through spin trapping/electron paramagnetic resonance (EPR) spectroscopy has been one of the great challenges in studying free radicals in biology. While 5-carboxy-5-methyl-1 pyrroline N-oxide, can specifically spin trap HO* in homogeneous solutions, the ionic nature of nitrone at physiologic pH prevents its entry into cells. We hypothesized that conversion of carboxyl-bearing spin probes such as nitrone into an esterase-hydrolyzable labile ester would permit intracellular localization and accumulation of the spin probes. To test the feasibility of such an approach, we prepared the model compound, 3-acetoxymethoxycarbonyl-2,2,5,5-tetramethyl-1 pyrrolidinyloxyl. This ester enabled ready accumulation of spin label to mM levels in lymphocytes. We suggest that its retention within these cells was the result of intracellular hydrolysis to 3-carboxy-2,2,5,5-tetramethyl-1 pyrrolidinyloxyl. Moreover, our studies show that aminoxyl was stable in the intracellular environment. These model studies suggest a viable strategy for detecting intracellular HO* by using the acetoxymethyl ester of 5-carboxy-5 methyl-1-pyrroline N-oxide. PMID- 14737667 TI - Understanding the puzzling chemistry of bicyclo[2.1.0]pentane. AB - Three thermal reactions of bicyclo[2.1.0]pentane have been studied by CASPT2-g3 and CASSCF electronic structure calculations. They are isomerization to cyclopentene, isomerization to 1,4-pentadiene, and cycloaddition to fumaronitrile. All three of these reactions exhibit unusual features that have prompted mechanistic debate. The present computational results provide a basis for understanding the experimental observations. PMID- 14737668 TI - Direct synthesis of unprotected phenols using palladium-catalysed cross coupling reactions of functionalised organozinc reagents. AB - Palladium-catalysed reaction of unprotected 2-, 3-, and 4-iodophenols with a range of amino acid derived organozinc reagents (not used in excess) gives the expected products in good to excellent yield, demonstrating that carbon-zinc bonds are not protonated by acidic phenols under the conditions of palladium catalysed coupling reactions. PMID- 14737669 TI - NMR and UV studies of 3'-S-phosphorothiolate modified DNA in a DNA : RNA hybrid dodecamer duplex; implications for antisense drug design. AB - High-resolution NMR spectroscopy has been used to establish the conformational consequences of the introduction of a single 3[prime or minute]-S phosphorothiolate link in the DNA strand of a DNA : RNA hybrid. These systems are of interest as potential antisense therapeutic agents. Previous studies on similarly modified dinucleotides have shown that the conformation of the sugar to which the sulfur is attached shifts to the north (C(3[prime or minute]) endo/C(2[prime or minute])-exo). Comparisons made between NOESY cross-peak intensities, and coupling constants from PE-COSY spectra, for both non-modified and modified duplexes confirm that this conformational shift is also present in the double helical oligonucleotide system. In addition it is noted that in both the dinucleotides and the modified duplex, the conformation of the sugar ring 3[prime or minute] to the site of modification is also shifted to the north. That this pattern is observed in the small monomeric system as well as the larger double helix is suggestive of some pre-ordering of the sequences. The conclusion is supported by consideration of the (1)H chemical shifts of the heterocyclic bases near the site of the modification. The enhanced stability that these conformational changes should bring was confirmed by UV thermal melting studies. Subsequently a series of singly and doubly 3[prime or minute]-S-phosphorothiolate modified duplexes were investigated by UV. The results are indicative of an additive effect of the modification with thermodynamic benefit being derived from alternate spacing of two modified linkers. PMID- 14737670 TI - Synthesis and cytotoxicity of 9-(2-deoxy-2-alkyldithio-beta-D arabinofuranosyl)purine nucleosides which are stable precursors to potential mechanistic probes of ribonucleotide reductases. AB - A series of 2[prime or minute]-thionucleosides, as potential inhibitors of ribonucleotide reductases, has been synthesized. Treatment of the 3[prime or minute],5[prime or minute]-O-TPDS-2[prime or minute]-O (trifluoromethanesulfonyl)adenosine with potassium thioacetate gave the arabino epimer of 2[prime or minute]-S-acetyl-2[prime or minute]-thioadenosine which was deacetylated to give 9-(3,5-O-TPDS-2-thio-[small beta]-d-arabinofuranosyl)adenine in high yield. Treatment of the latter with diethyl azodicarboxylate-C(3)H(7)SH THF gave 2[prime or minute]-propyl disulfide which was desilylated to give 9-(2 deoxy-2-propyldithio-[small beta]-d-arabinofuranosyl)adenine. Subsequent tosylation (O5[prime or minute]) and displacement of the tosylate with pyrophosphate afforded the 5[prime or minute]-O-diphosphate in a stable form as propyl mixed-disulfide, which upon treatment with dithiothreitol releases 9-(2 thio-[small beta]-d-arabinofuranosyl)adenine 5[prime or minute]-diphosphate. The arabino 2[prime or minute]-mercapto group might interact with the crucial thiyl radical at cysteine 439 leading to the inhibition of ribonucleotide reductases via formation of a Cys439-2[prime or minute]-mercapto disulfide bridge. The 2,6 diamino-, 2-amino-6-chloro- and 2-amino-6-methoxypurine ribosides were also converted to the corresponding 2[prime or minute]-deoxy-2[prime or minute] propyldithio-[small beta]-d-arabinofuranosyl nucleosides, which might serve as convenient precursors to the arabino epimer of 2[prime or minute]-thioguanosine. Analogously, 2[prime or minute]-deoxy-2[prime or minute]-propyldithioadenosine was prepared from 9-([small beta]-d-arabinofuranosyl)adenine. The nucleoside disulfides show modest cytotoxicity in a panel of human tumor cell lines. PMID- 14737671 TI - Conformation-activity relationships in polyketide natural products. Towards the biologically active conformation of epothilone. AB - The conformation-activity relationships for the biologically active polyketide, epothilone, have been determined. Computer-based molecular modeling and high field NMR techniques have provided the solution preferences for epothilones and. For the C1-C8 polypropionate region, two conformational families, conformers 1 and 2, have been identified as having significant populations in polar and non polar solvents. In the C11-C15 region, additional flexibility was observed and two local conformations have been identified as important, conformers 3 and 4. Epothilone analogues with altered conformational profiles have been designed and synthesized. Conformational analysis and the results of biological assays have been correlated to provide increased understanding of the biologically active conformation for the epothilone class of natural product. Conformation-activity relationships have been shown to be an important complement to structure-activity data. PMID- 14737672 TI - Application of Fmoc-amino acid carrying an unmasked carbohydrate to the synthesis of the epidermal growth factor-like domain of bovine blood coagulation factor IX. AB - The epidermal growth factor (EGF)-like domain of the bovine blood coagulation factor IX (45-87) carrying glucose at Ser(53) was synthesized by a solid-phase method using 9-fluorenylmethoxycarbonyl (Fmoc)-amino acids. The introduction of Ser(53) was carried out using the benzotriazolyl ester of Fmoc-serine carrying an unmasked glucose. The remaining sequence was also introduced using the benzotriazolyl ester. HPLC analysis of the crude peptide shows that acylation of the free hydroxyl group of the glucose was not significant, demonstrating that the amino acid carrying an unmasked carbohydrate is a useful building block for solid-phase synthesis. PMID- 14737673 TI - An organic hydrogel as a matrix for the growth of calcite crystals. AB - The growth of calcite in an aqueous gel of was studied and the appearance of the crystals was found to change over time. Crystals removed from the gel at progressively longer times showed severely affected surfaces resulting from dissolution. If crystals were removed from the gel after 3.5 hours, at which point there were no etch pits, and then placed in either buffer or pure water, etch pits, similar to those observed on crystals that are left in the gel, were observed. Control calcite crystals exposed to similar conditions (water or buffer) show no significant dissolution after equivalent times. A probable cause of the altered dissolution is the non-specific occlusion of gelator aggregates at sites of imperfection. The gel appears to provide a microenvironment in which the molecules that form the matrix also participate in the crystallization. This system allows the study of the unique properties of a gel for influencing the nucleation and growth of inorganic crystals, some of which may be important for better understanding biomineralization. PMID- 14737698 TI - Revision rhinoplasty. AB - Revision rhinoplasty can be one of the most complicated procedures performed by the facial plastic surgeon. As septal cartilage is often not available in revision procedures, grafting material is often needed. This material can come in the form of autogenous bone and cartilage. Allografts also can be used, including mersilene, expanded polytetrafluoroethylene, and porous high-density polyethylene (PHDPE). In this article, emphasis is placed on the senior author's method in evaluating candidates for revision rhinoplasty as well as techniques using PHDPE. In addition, the properties of the more commonly used allografts are described, including the advantages and disadvantages of using each material in revision rhinoplasty procedures. In comparing the various alloplastic materials available, it is shown that PHDPE has properties that make it an excellent implant for revision rhinoplasty. PMID- 14737674 TI - Synthetic anisomycin analogues activating the JNK/SAPK1 and p38/SAPK2 pathways. AB - The synthesis of C(4)H and C(4)Me analogues of the JNK/p38 pathway activator anisomycin, based upon an aldol or Claisen construction of the C(3)-C(4) bond, has been demonstrated. The relative activation of the JNK/SAPK1 and p38/SAPK2 pathways in RAW macrophages by these analogues, and their synthetic precursors, has been assessed using immunoblot assays against phosphorylated c-Jun and MAPKAP K2. These studies demonstrate that some of the synthetic C(4) analogues are also potent activators of these stress kinase pathways. PMID- 14737699 TI - Rhinoplasty in the aging nose. AB - The aging nose presents a difficult challenge for the rhinoplastic surgeon. To best address the patient's wishes, the surgeon must possess a sound understanding of the atrophic changes associated with the aging nose, including tip ptosis, increased nasal bulbosity, a lengthened nasal appearance, and altered nasal airflow patterns. Surgical emphasis is placed on conservative structured reduction to address functional concerns. Overresection risks destabilizing the nasal tip and altering the patient's concrete self-image. This article outlines our approach to rhinoplasty in the aging nose with a special emphasis on the use of porous polyethylene alloimplants to provide added structural support. PMID- 14737700 TI - Reduction structured rhinoplasty. AB - Rhinoplastic surgeons continue to seek technical refinements that will result in a consistently reliable postoperative outcome. Over the past 25 years there has been a steady progression away from the simple Joseph reductive rhinoplasty technique toward the use of various grafts to improve both the functional outcomes and aesthetic end result of this procedure. The original reductive template continues to be a major component of a new conceptual paradigm, which has evolved to incorporate the understanding that structural components must be placed to allow a more precise outcome. Specifically, once the nose is reduced to proportions that correspond to the preoperative analysis, separate grafts and implants are placed to prevent a change in the shape of the nose by the constricting effects of the skin shrink-wrapping around the skeletal framework. This article focuses on the use of implants for structural architecture in the senior author's primary reduction-structured rhinoplasty cases. PMID- 14737701 TI - Nasal dorsal augmentation with silicone implants. AB - Silicone rubber has been used safely and effectively for facial augmentation for nearly 5 decades in eastern Asia. We have used silicone rubber nasal implants in primary ethnic rhinoplasty and have found consistent and long-lasting results with low complication rates. Silicone dorsal nasal augmentation in primary rhinoplasty avoids donor site morbidity and implant resorption as seen with autogenous implants. Silicone nasal implants have a low extrusion and infection rate. In the appropriate patient with proper placement, silicone nasal implant is nearly the ideal implant material. PMID- 14737702 TI - Expanded polytetrafluoroethylene implants in rhinoplasty: literature review, operative techniques, and outcome. AB - Gore-Tex, a form of expanded polytetrafluoroethylene (ePTFE), over the past 30 years has attracted much attention as an alloplast for use in rhinoplasty, both from advocates and opponents of its use. It has many desirable traits as an alloplast implant, but many surgeons harbor hesitation and reluctance for alloplast use in rhinoplasty based on historical data of previous nasal implants. Only when objective data from large series of patients with long-term follow-up become available will such skepticism be resolved. Large series of patients with Gore-Tex implant placement during rhinoplasty are beginning to emerge in the literature. The purpose of this article is twofold. The first is to provide the reader with an up-to-date review of the literature on the host response to polymer implants and, second, of the current indications and operative techniques for use and outcomes of Gore-Tex implants in rhinoplasty. PMID- 14737703 TI - Management of the severe bulbous nasal tip using porous polyethylene alloimplants. AB - Reliable outcomes in correction of the severely bulbous nasal tip remain elusive. Reduction and binding of the tip cartilages result in a partial external nasal valve collapse. If a concurrent dorsal hump reduction is accomplished and osteotomies are completed to close the open roof deformity, then a severe constriction of the external valve may result. This is best avoided by resection of the residual lower lateral cartilages and substitution with external valve batten implants. The tripod of nasal tip supports is then rigidly reconstituted. When autogenous graft materials are considered inadequate or undesirable, the surgeon may safely use porous polyethylene alloimplants to support the external nasal valve. A dependable surgical technique based on extensive experience with these materials is outlined. PMID- 14737704 TI - Adjustment of subtle postoperative nasal defects: managing the "near-miss" rhinoplasty. AB - Rhinoplasty entails several steps that affect the appearance and function of the nose in an interrelated way. Occasionally, an excellent result can be marred by a minor imperfection and lead to dissatisfaction of both patient and surgeon. For significant pathology, revision rhinoplasty can be performed safely if the architecture and physiology of the nose are respected. To correct the "near-miss" result, minor corrections can successfully restore the aesthetic result. This can require a different frame of reference and approach and can often be thought of in terms of soft tissue correction. Subcutaneous fibrosis, skin depressions, asymmetries, and deficits can be corrected with minor procedures and techniques not routinely associated with rhinoplasty. We detail several methods that can be used to restore simply and effectively an otherwise excellent rhinoplasty result. PMID- 14737705 TI - Sclerosing lipogranuloma of the nose: a new treatment using adipose tissue transplantation. AB - We report a series of 45 patients visiting our clinic with distorted contour of the nose after having their noses augmented with injectable substances. The most common clinical presentation was a palpable mass on the dorsum of the nose, erythema with or without telangiectasia on the overlying skin, and nodularity of the nose. The onset of symptoms varied from 1 to 16 years after injection. They were treated by surgical excision, but this was always followed by a saddle-nose deformity. To correct this defect, we propose a new technique of adipose tissue transplantation that yields satisfactory results. Pathological reports of the excised mass showed a foreign body granuloma known as a sclerosing lipogranuloma (paraffinoma or siliconoma). PMID- 14737706 TI - Secondary cleft-lip rhinoplasty utilizing porous high-density polyethylene. AB - Secondary nasal deformity associated with cleft lip is a difficult surgical task. For more than 100 years, numerous surgical methods have been created to address the structural changes that occur over time after the primary surgery. Although early surgical methods have relied on various rotational flaps and local tissue manipulation, the recent advent of synthetic materials has provided facial plastic surgeons with more tools to correct the structural and supportive deficiencies. In this article, we describe our 10-year experience with porous high-density polyethylene (PHDPE) implants in secondary cleft-lip rhinoplasty. PHDPE implants have been well tolerated and achieved excellent long-term aesthetic results through fibrovascular ingrowth to the surrounding tissue. PMID- 14737707 TI - Molecular systematic analysis reveals cryptic tertiary diversification of a widespread tropical rain forest tree. AB - The broad geographic range of many Neotropical rain forest tree species implies excellent dispersal abilities or range establishment that preceded the formation of current dispersal barriers. In order to initiate historical analyses of such widespread Neotropical trees, we sequenced the nuclear ribosomal spacer (ITS) region of Symphonia globulifera L. f. (Clusiaceae) from populations spanning the Neotropics and western Africa. This rain forest tree has left unmistakable Miocene fossils in Mesoamerica (15.5-18.2 Ma) and in South America ( approximately 15 Ma). Although marine dispersal of S. globulifera is considered improbable, our study establishes three marine dispersal events leading to the colonization of Mesoamerica, the Amazon basin, and the West Indies, thus supporting the paleontological data. Our phylogeographic analysis revealed the spatial extent of the three Neotropical S. globulifera clades, which represent trans-Andes (Mesoamerica+west Ecuador), cis-Andes (Amazonia+Guiana), and the West Indies. Strong phylogeographic structure found among trans-Andean populations of S. globulifera stands in contrast to an absence of ITS nucleotide variation across the Amazon basin and indicates profound regional differences in the demographic history of this rain forest tree. Drawing from these results, we provide a historical biogeographic hypothesis to account for differences in the patterns of beta diversity within Mesoamerican and Amazonian forests. PMID- 14737708 TI - Carotenoids, immunocompetence, and the information content of sexual colors: an experimental test. AB - Many male birds use carotenoid pigments to acquire brilliant colors that advertise their health and condition to prospective mates. The direct means by which the most colorful males achieve superior health has been debated, however. One hypothesis, based on studies of carotenoids as antioxidants in humans and other animals, is that carotenoids directly boost the immune system of colorful birds. We studied the relationship between carotenoid pigments, immune function, and sexual coloration in zebra finches (Taeniopygia guttata), a species in which males incorporate carotenoid pigments into their beak to attract mates. We tested the hypotheses that increased dietary carotenoid intake enhances immunocompetence in male zebra finches and that levels of carotenoids circulating in blood, which also determine beak coloration, directly predict the immune response of individuals. We experimentally supplemented captive finches with two common dietary carotenoid pigments (lutein and zeaxanthin) and measured cell-mediated and humoral immunity a month later. Supplemented males showed elevated blood carotenoid levels, brighter beak coloration, and increased cell-mediated and humoral immune responses than did controls. Cell-mediated responses were predicted directly by changes in beak color and plasma carotenoid concentration of individual birds. These experimental findings suggest that carotenoid-based color signals in birds may directly signal male health via the immunostimulatory action of ingested and circulated carotenoid pigments. PMID- 14737709 TI - Inferring process from pattern in plant invasions: a semimechanistic model incorporating propagule pressure and environmental factors. AB - Propagule pressure is intuitively a key factor in biological invasions: increased availability of propagules increases the chances of establishment, persistence, naturalization, and invasion. The role of propagule pressure relative to disturbance and various environmental factors is, however, difficult to quantify. We explored the relative importance of factors driving invasions using detailed data on the distribution and percentage cover of alien tree species on South Africa's Agulhas Plain (2,160 km2). Classification trees based on geology, climate, land use, and topography adequately explained distribution but not abundance (canopy cover) of three widespread invasive species (Acacia cyclops, Acacia saligna, and Pinus pinaster). A semimechanistic model was then developed to quantify the roles of propagule pressure and environmental heterogeneity in structuring invasion patterns. The intensity of propagule pressure (approximated by the distance from putative invasion foci) was a much better predictor of canopy cover than any environmental factor that was considered. The influence of environmental factors was then assessed on the residuals of the first model to determine how propagule pressure interacts with environmental factors. The mediating effect of environmental factors was species specific. Models combining propagule pressure and environmental factors successfully predicted more than 70% of the variation in canopy cover for each species. PMID- 14737710 TI - Flammability is a niche construction trait: canopy architecture affects fire intensity. AB - By affecting local fire intensities or the probability of ignition, traits that influence plant flammability may indirectly control selection for fire-related life-history and physiological traits. The retention of dead branches in the canopy has been cited as contributing to plant flammability. No experiment, however, has demonstrated that differences in plant canopy architecture on the scale of observed variation in nature can affect local fire characteristics. I experimentally manipulated canopies of Adenostoma fasciculatum, a California shrub that naturally retains dead branches, to mimic degrees of self-pruning in four small-scale (4 m x 6 m) treatments: removal of all canopy dead wood, clipping of all dead wood with wood left as litter, an unmanipulated treatment, and a dead wood addition. Treatment plots were burned in large-scale prescribed fires. Fire temperatures and heat release were significantly higher in Unmanipulated and Addition treatments, demonstrating a significant local effect of dead branch retention. Removal and Clip and Leave treatments did not differ significantly; the observed effect is a result of canopy architecture rather than differences in total fuel load. PMID- 14737711 TI - Partitioning species diversity across landscapes and regions: a hierarchical analysis of alpha, beta, and gamma diversity. AB - Species diversity may be additively partitioned within and among samples (alpha and beta diversity) from hierarchically scaled studies to assess the proportion of the total diversity (gamma) found in different habitats, landscapes, or regions. We developed a statistical approach for testing null hypotheses that observed partitions of species richness or diversity indices differed from those expected by chance, and we illustrate these tests using data from a hierarchical study of forest-canopy beetles. Two null hypotheses were implemented using individual- and sample-based randomization tests to generate null distributions for alpha and beta components of diversity at multiple sampling scales. The two tests differed in their null distributions and power to detect statistically significant diversity components. Individual-based randomization was more powerful at all hierarchical levels and was sensitive to departures between observed and null partitions due to intraspecific aggregation of individuals. Sample-based randomization had less power but still may be useful for determining whether different habitats show a higher degree of differentiation in species diversity compared with random samples from the landscape. Null hypothesis tests provide a basis for inferences on partitions of species richness or diversity indices at multiple sampling levels, thereby increasing our understanding of how alpha and beta diversity change across spatial scales. PMID- 14737712 TI - Frequency-dependent inbreeding depression in Amsinckia. AB - If the competitive ability of plants produced by self-pollination differs from that of plants derived by outcrossing, then the magnitude of inbreeding depression may be influenced by the composition of the competitive environment (i.e., the frequency of plants that have arisen from selfing and outcrossing in the neighborhood of "target" plants in which inbreeding depression is expressed). Here, we report the results of experiments designed to examine whether inbreeding depression is influenced by the frequency of inbred plants in the competitive neighborhood. Two species of the annual plant genus Amsinckia were studied, one a near-complete selfer (Amsinckia gloriosa) and the other a partial outcrosser (Amsinckia douglasiana). Competition experiments were conducted in artificial stands composed of different mixtures of inbred and outbred progeny. The fitnesses of progeny were found to be significantly influenced by the composition of the competing neighborhood. The fitness of target plants, however, did not vary monotonically with the frequency of inbred plants in the neighborhood. Rather, for A. gloriosa, maximum performance was observed when there was an intermediate frequency of inbred neighbors. For A. douglasiana, the opposite pattern was found. The results suggest that competition among progeny has the potential to play a role in the selection of self-fertilization and possibly in the maintenance of mixed mating systems. PMID- 14737713 TI - Talkin' 'bout my generation: environmental variability and cohort effects. AB - In variable environments, it is probable that environmental conditions in the past can influence demographic performance now. Cohort effects occur when these delayed life-history effects are synchronized among groups of individuals in a population. Here we show how plasticity in density-dependent demographic traits throughout the life cycle can lead to cohort effects and that there can be substantial population dynamic consequences of these effects. We show experimentally that density and food conditions early in development can influence subsequent juvenile life-history traits. We also show that conditions early in development can interact with conditions at maturity to shape future adult performance. In fact, conditions such as food availability and density at maturity, like conditions early in development, can generate cohort effects in mature stages. Based on these data, and on current theory about the effects of plasticity generated by historical environments, we make predictions about the consequences of such changes on density-dependent demography and on mite population dynamics. We use a stochastic cohort effects model to generate a range of population dynamics. In accordance with the theory, we find the predicted changes in the strength of density dependence and associated changes in population dynamics and population variability. PMID- 14737714 TI - The forager's dilemma: food sharing and food defense as risk-sensitive foraging options. AB - Although many variants of the hawk-dove game predict the frequency at which group foraging animals should compete aggressively, none of them can explain why a large number of group foraging animals share food clumps without any overt aggression. One reason for this shortcoming is that hawk-dove games typically consider only a single contest, while most group foraging situations involve opponents that interact repeatedly over discovered food clumps. The present iterated hawk-dove game predicts that in situations that are analogous to a prisoner's dilemma, animals should share the resources without aggression, provided that the number of simultaneously available food clumps is sufficiently large and the number of competitors is relatively small. However, given that the expected gain of an aggressive animal is more variable than the gain expected by nonaggressive individuals, the predicted effect of the number of food items in a clump-clump richness-depends on whether only the mean or both the mean and variability associated with payoffs are considered. More precisely, the deterministic game predicts that aggression should increase with clump richness, whereas the stochastic risk-sensitive game predicts that the frequency of encounters resulting in aggression should peak at intermediate clump richnesses or decrease with increasing clump richness if animals show sensitivity to the variance or coefficient of variation, respectively. PMID- 14737715 TI - The effects of enrichment on the dynamics of apparent competitive interactions in stage-structured systems. AB - In the absence of other limiting factors, assemblages in which species share a common, effective natural enemy are not expected to persist. Although a variety of mechanisms have been postulated to explain the coexistence of species that share natural enemies, the role of productivity gradients has not been explored in detail. Here, we examine how enrichment can affect the outcome of apparent competition. We develop a structured resource/consumer/natural enemy model in which the prey are exposed to attacks during a vulnerable life phase, the length of which depends on resource availability. With a single prey species, the model exhibits the "paradox of enrichment," with unstable dynamics at high levels of resource productivity. We extend this model to consider two prey species linked by a shared predator, each with their own distinct resource base. We derive invasion and stability conditions and examine how enrichment influences prey species exclusion and coexistence. Contrary to expectations from simpler, prey dependent models, apparent competition is not necessarily strong at high productivity, and prey species coexistence may thus be more likely in enriched environments. Further, the coexistence of apparent competitors may be facilitated by unstable dynamics. These results contrast with the standard theory that apparent competition in productive environments leads to nonpersistent interactions and that coexistence of multispecies interactions is more likely under equilibrial conditions. PMID- 14737716 TI - Fitness components versus total demographic effects: evaluating herbivore impacts on a perennial herb. AB - I examined the long-term effects of cryptic mollusk herbivory on seven fitness components in the perennial herb Lathyrus vernus and also calculated measures of total fitness effects. Natural correlations and experimental exclusion of mollusks showed that herbivory is associated with an increased probability of dying or staying dormant, poorer growth, and a lower probability of flowering. The average yearly reduction in population growth rate (lambda) caused by mollusks in the experiment was 0.14. The largest contribution to this decrease in total fitness occurred through a decreased survival of established plants. In contrast, seedling emergence and probability of flowering were the fitness components that were most affected in terms of relative change. The more important a life-cycle transition was for population growth rate in terms of its elasticity, the less it was affected by herbivore damage. These results suggest that simple analyses of the magnitudes of effects on individual components of plant performance are poor predictors of the magnitude of total fitness effects and tolerance to herbivory. This is because total fitness is differently sensitive to different phases of the life cycle and because plants strive to maintain the functions most important to fitness. PMID- 14737717 TI - Asymmetric mandibles of water-scavenger larvae improve feeding effectiveness on right-handed snails. PMID- 14737718 TI - Measuring nonlinear selection. PMID- 14737719 TI - Is parental care the key to understanding endothermy in birds and mammals? PMID- 14737720 TI - Reproduction: the adaptive significance of endothermy. PMID- 14737722 TI - Uptake of radiolabelled herceptin by experimental mammary adenocarcinoma. AB - BACKGROUND: The aim of this study was to examine the biodistribution of (131)I herceptin in C3H/Bi mice with transplantable mammary adenocarcinoma with a high frequency of C-erbB2 receptor expression. MATERIAL AND METHODS: Mice C3H/Bi with subcutaneously transplanted mammary adenocarcinoma were used as animal model to study the interaction between C-erbB2 receptor and hercepin, a humanized anti-C erbB2 monoclonal antibody. The expression of the gene encoding C-erbB2 receptor in the tumours was studied by the RT-PCR technique. RESULTS: Expression of this gene was found in 66% of the studied cases. Similarly, the presence of the C erbB2 receptor in 77% of the tumours was detected by a Western blot analysis with the use of herceptin. Biodistribution experiments of iodine-labelled herceptin in mice C3H/Bi with adenocarcinoma revealed its maximal accumulation in the tumours at 48 hours since the i.v. injection (7% ID/g). The tumour/muscle radioactivity ratio reached its highest value (above 20) also at 48 hours after the injection. CONCLUSIONS: C3H/Bi mice with this adenocarcinoma may be a good experimental model to study herceptin, or its fragments, labelled with different radionuclids for preliminary evaluation of their usefulness in the therapeutic and diagnostic aspects of breast cancer. PMID- 14737723 TI - Development of radioimmunoassay for the measurement of human leptin in serum. AB - BACKGROUND: Leptin is a 16 kDa polypeptide hormone encoded by the obese gene (ob) and secreted by adipose tissue. This hormone plays a major role in energy homeostasis and regulation of food intake and body weight. It also affects the metabolic, neuroendocrine and reproductive systems. MATERIAL AND METHODS: Labelling of recombinant human leptin with (125)I was best performed by the Chloramine-T method. New Zealand white rabbits were immunised with recombinant human leptin, cross-reaction of obtained antisera was analyzed with 10 different antigens. The separation of bound and free fractions was performed using the second antibody - PEG method. RESULTS: The obtained tracer had specific activities of 2.8-3.3 kBq/ microg and had a stability of 5 weeks. A highly specific polyclonal antibody was obtained without measurable cross-reaction against the analysed antigens. Concentrations of human leptin were measured by a single overnight incubation assay with a sensitivity of 0.5 ng/ml and a measuring range of 0.5-100 ng/ml. The intra-assay and inter-assay coefficient of variation was under 6% and 8%, respectively. Recovery ranged from 88% to 106%. CONCLUSIONS: Serum human leptin concentrations can be accurately and precisely measured by this new radioimmunoassay. Preliminary results obtained from the measurement of serum leptin in lean, overweight and obese patients are presented. Serum leptin concentrations correlated with body mass index and were significantly higher in women than in men, except for obese patients. PMID- 14737724 TI - Application of artificial neural network algorithm to detection of parathyroid adenoma. AB - BACKGROUND: The most common radionuclide procedures for parathyroid imaging are (99m)Tc-MIBI/ (99m)Tc pertechnetate subtraction scintigraphy and (99m)Tc-MIBI double-phase imaging, with estimation of MIBI wash-out rate. Those two methods are by some authors regarded as complementary techniques, yielding the best evaluation of parathyroid gland if performed conjointly. By such an approach it seems reasonable to substitute the visual assessment of neck scintigrams and semiquantitative evaluation of MIBI wash-out rate with a single, common procedure. The aim of this study was application of the Artificial Neural Network (ANN) simulated by the computer program to detection and localisation of pathological parathyroid tissue in the planar neck scintigrams. MATERIAL AND METHODS: The applied algorithm was based on simultaneous data processing in sets of 3 single pixels, each of them belonging to one of the three consecutive neck scintigrams generated 20 min. after (99m)TcO(4 )- administration, 10 min. after (99m)Tc-MIBI injection and 120 min. after (99m)Tc-MIBI injection, respectively. Those scintigrams were aligned which each other according to the same vertical and horizontal co-ordinates. The training patterns were obtained from 25 patients by searching for maximum count numbers within small ROIs drawn in selected scintigraphic areas, arbitrarily classified and coded in a numerical scale. In 10 pts the results of ANN simulation were compared with those obtained by common conventional assessment of two radionuclide parathyroid examinations: subtraction method and (99m)Tc-MIBI double-phase imaging. RESULTS: The training patterns processed by the neural network showed a close relationship with the results of visual assessment of original neck scintigrams, with R square coefficient R(2) = 0.717, and standard error equal to 0.243. Similar comparison between original data and results of multidimensional regression analysis yielded weaker relationship, with R(2) = 0.543 and standard error 0.567. Parametric images obtained by the neural network presented regions with homogeneously distributed, relatively high activity, greater than or equal to 750 cts/pixel, visualized in areas of confirmed abnormal parathyroid location. In all 10 patients with suspected parathyroid adenoma results obtained by ANN simulation agreed with those by conventional methods. In five of these cases no parathyroid abnormalities were found. In the remaining 5 subjects results of both approaches were positive but the abnormalities were depicted more distinctly and visualised more clearly in parametric images received by ANN than in original scans. CONCLUSIONS: Application of trained ANN enables objective and quantitative detection and localisation of parathyroid adenoma and is a good alternative for conventional radionuclide imaging procedures used in diagnosing parathyroid abnormality. Including in neural network simulation not only scintigraphic data, but also clinical symptoms and/or some other indicators of parathyroid abnormality, parathormone level first of all, should be a next step in developing a procedure for assessing parathyroid abnormality, of high diagnostic accuracy. PMID- 14737725 TI - Diagnostic sensitivity of two radio receptor assays (TRAK Assay and TRAK Dyno Human) for detection of TSH receptor antibodies. AB - Radio receptor assays for the detection of TSH receptor antibodies in serum are typically based on binding the competition of TSH-R antibodies and (125)I "labeled" TSH for membrane preparation of thyrocytes (TBII tests). The sensitivity of the available tests utilizing porcine cell membranes was found to be around 80%. A new test (TRAK Dyno human, BRAHMS) utilizes human recombinant TSH receptor and human standard material that is supposed to improve the performance of the test. We have compared the results of these two assays. The sensitivity of the TRAK Assay tested in 356 patients with untreated Graves' disease was found to be 85%, and 97.5% for TRAK Dyno human in 111 newly diagnosed patients. Both tests were performed from the same serum specimen for 60 of the investigated patients. The TRAK Assay was positive in 50 patients (83.2%) and TRAK Dyno human in 59 patients (98.3%). The specificity of the new radio receptor assay was also improved. PMID- 14737726 TI - Radioidine therapy temporarily increases circulating endothelial cells and decreases endothelial progenitor cells. AB - BACKGROUND: Radiotherapy can cause vascular injury. No data on radioiodine therapy and vascular damage are available. MATERIAL AND METHODS: We examined the number of circulating endothelial cells (CEC) and circulating endothelial progenitor cells (CEPC) before therapy 1, 2, 3 and 5 days as well as 1, 2, 3, 4, 6, 8, 10, and 12 weeks after therapy with (131)I at doses ranging from 5-200 mCi. The individual number of CEC and CEPC is associated with the presence of risk factors. RESULTS: Irrespective of prevalues, CEC exhibited a significant dose dependent temporary increase reaching the maximum in weeks 1 and 2. In contrast, CEPC show a decrease at the same time. CONCLUSIONS: These results indicate that (131)I-therapy induces a dose-dependent radiation injury at the vascular wall level enhancing endothelial desquamation and reducing reendothelialization and thereby a proatherogenic stage. The clinical consequence of these findings still needs to be assessed. PMID- 14737727 TI - Use of Tc99m-nanocolloid for sentinel nodes identification in cervical cancer. AB - BACKGROUND: The initial draining lymph node for a primary tumor is referred to as the "sentinel" node. Firstly adopted in the management of patients with cutaneous melanoma and breast cancer, it is now widely tested in cervical cancer. In patients with cervical cancer, lymph node status is the most important prognostic factor for survival. In patients with cervical cancer FIGO stage I and II pelvic lymph node metastases are expected in 0-16 and 24.5-31% and para-aortic lymph node metastases are expected in 0-22 and 11-19% of patients. The removal of pelvic and para-aortic lymph nodes is essential for assessing the biology of the disease. Lymphoscintigraphy enables the visualisation of lymphatic drainage patterns from a great variety of tumour sites prior to surgery. Therefore, the current procedure is to perform the pre-operative mapping of sentinel nodes by static and/or dynamic lymphoscintigraphy, followed by in vivo identification using a gamma detection probe and selective surgical resection. MATERIAL AND METHODS: Between 2001-2003, 37 patients with cervical cancer FIGO stage I-IIa were seemed to be qualified to undergo lymphoscintigraphy. The day before surgery (99m)Tc-nanocolloid (100 MBq; 0.5-1.0 ml in volume) was applied in each quadrant of the cervix or around the tumor. The static scintigraphic scans were performed after 2 hours p.i. using a dual-head large-field-of-view Siemens gamma-camera equipped with high resolution collimators. SNs were identified intra-operatively using a handheld gamma detection probe (Navigator GPS-Tyco) and intra-operative lymphatic mapping with blue dye. After a resection of the SNs, a standard radical hysterectomy with pelvic and low para-aortic lymph node dissection was performed. Tumor characteristics were compared with sentinel node detection and with the histopathological and immunohistochemical results. RESULTS: The scintigraphy showed a focal uptake in 35 of the 37 patients. In all women one or more sentinel lymph nodes were identified intra-operatively. Of them, 24 patients had those located bilaterally. Histologically positive SNs were found in 5 women (13.5%). CONCLUSIONS: A combination pre-operatively administered radioactively labelled albumin with blue dye allows the successful detection of SN in patient with cervical cancer. This technique will result in a real advance in the less aggressive management of patients with early stage cervical cancer. Sentinel lymph node status may be representative of the pelvic lymph nodes status in cervical cancer and thus could provide important information for further treatment. PMID- 14737728 TI - The influence of depth of marker administration on sentinel node detection in cervical cancer. AB - BACKGROUND: Regional lymph node surgical management is an integral part of cervical cancer therapy. In gynaecological oncology, recent studies have confirmed the utility of the sentinel node concept in vulvar and cervical cancer. The method of the marker's administration is considered to play an important role in sentinel node detection. MATERIAL AND METHODS: 60 patients with cervical cancer (stage IB-IIA) underwent SLN detection during radical abdominal hysterectomy. The patients were randomly divided into two groups: the first group of 30 patients with 0.5-1cm deep marker injection, the second with sub-epithelial marker injection. Gamma-camera scanning, as well as hand-held probe detection was applied. RESULTS: All hot nodes visualised on lymphoscintigraphy were "hot" when using the hand-held gamma probe. Deep marker injection revealed a sentinel node in 27 patients (90%) on both sides, in 3 patients (10%) only on one side. Only 40 (67%) sentinel nodes were blue-stained. Sub-epithelial marker administration revealed a sentinel node on both sides in all 30 patients (100%). In 28 patients (93.3%) the sentinel nodes were radioactive and blue-stained, in one case not blue stained on either side, in one case blue stained only on one side. CONCLUSIONS: The sentinel node detection rate in cervical cancer is relatively high and depends on the applied technique. The superficial administration of radiocolloid and the blue dye into the cervix provides a higher sentinel node detection rate than deep administration in cervical cancer patients. PMID- 14737729 TI - The Nuclear Medicine Technologist in Europe - current status and future perspective. PMID- 14737730 TI - FDG PET and alternative imaging in the management of thyroid carcinoma. AB - Differentiated carcinoma of the thyroid are one of rare malignancies that is associated with excellent prognosis. Follow-up with regular thyroglobulin assay and (131)I whole-body scan is capable of detecting residual or recurrent disease with great sensitivity and specificity. However, there is overwhelming evidence to suggest that this approach is not fail-safe due to increasing reports of false negative and false positive results, which may result in missed or unwarranted therapy with (131)I. This article will review the current management of differentiated carcinoma of the thyroid and the possible causes of the reported inadequacy of thyroglobulin and (131)I whole-body scan to detect residual or recurrent disease, and the increasing role of alternative imaging, particularly (18)F-FDG PET in the management of this curable malignancy. PMID- 14737731 TI - Role of myocardial perfusion imaging in risk stratification. AB - This review paper is based on a topic invited for review presented by the first author at the Annual Meeting of Society of Nuclear Medicine of Serbia and Montenegro, September 2003. The optimal management of patients affected by coronary artery disease requires safe and cost-effective techniques for assessing the risk of subsequent cardiac events or the need for surgery. The ideal test should distinguish between high risk patients who will benefit from aggressive management and low risk patients who can be managed conservatively. Stress testing alone is believed to be inadequate for the assessment of risk. Nuclear Medicine techniques have routinely been used in the identification of patients with ischaemic heart disease and those with viable myocardium post myocardial infarction. While the essential methodology for the techniques remains same, the reporting and interpretation criteria for determining future risk are different. For instance, a fixed perfusion defect on stress myocardial perfusion imaging in a patient presenting cardiactype chest pain to the cardiologists for the first time, has a different value from a similar patient being presented to the noncardiologist for major vascular surgery. A review of the literature concerning the current usage of nuclear cardiology techniques in risk stratification is presented. PMID- 14737733 TI - Columnar cell thyroid carcinoma - diagnostic dilemmas and pitfalls. AB - BACKGROUND: Columnar cell carcinoma is a rare variant of papillary thyroid carcinoma associated with aggressive clinical behaviour. A CASE REPORT: of a 34 year-old male patient, who presented with the rapidly growing mass in the neck, extending to the anterior and middle mediastinum, tightly closing the upper thoracic apperture and causing tracheal and oesophageal deviation, with minimal compression and stenosis. A diagnosis of columnar cell carcinoma, arising from the ectopic thyroid tissue just adjacent to the left thyroid lobe was based on histological and intraoperative findings. Near total thyroidectomy and lymph-node dissection were followed by external beam radiotherapy of the neck and mediastinum, chemotherapy and radioiodine ablation of the remaining functional thyroid tissue. Pre-and post-operative radionuclide imaging (99mTc(V)-DMSA, 99mTc MIBI, 123-I-mIBG and Octreoscan findings are discussed, with a special emphasis given to the dilemmas in histological characterisation of the tumor, the problems in therapeutic approach and the dilemmas and pitfalls in the interpretation of radionuclide findings in this patient, especially the ones performed post radiotherapy of the neck and mediastinum. PMID- 14737741 TI - The vasodilatory actions of insulin on resistance and terminal arterioles and their impact on muscle glucose uptake. AB - Whether a discrete vascular action of insulin in skeletal muscle integrally participates in insulin-mediated glucose disposal has been extensively examined but remains a contentious issue. Here, we review some of the data both supporting and questioning the role of insulin-mediated increases in limb blood flow in glucose metabolism. We advance the hypothesis that controversy has arisen, at least in part, from a failure to recognize that insulin exerts at least three separate actions on the peripheral vasculature, each with its own characteristic dose and time responsiveness. We summarize how, viewed in this manner, certain points of contention can be resolved. We also advance the hypothesis that an action on the precapillary arteriole may play the dominant role in mediating perfusion-dependent effects of insulin on glucose metabolism in muscle. PMID- 14737742 TI - Progenitor cells in the adult pancreas. AB - The beta-cell mass in the adult pancreas possesses the ability to undergo limited regeneration following injury. Identifying the progenitor cells involved in this process and understanding the mechanisms leading to their maturation will open new avenues for the treatment of type 1 diabetes. However, despite steady advances in determining the molecular basis of early pancreatic development, the identification of pancreatic stem cells or beta-cell progenitors and the molecular mechanisms underlying beta-cell regeneration remain unclear. Recent advances in the directed differentiation of embryonic and adult stem cells has heightened interest in the possible application of stem cell therapy in the treatment of type 1 diabetes. Drawing on the expanding knowledge of pancreas development, beta-cell regeneration and stem cell research, this review focuses on progenitor cells in the adult pancreas as a potential source of beta-cells. PMID- 14737743 TI - The role of lipids and protein kinase Cs in the pathogenesis of diabetic retinopathy. AB - Diabetic retinopathy is one of the most common complications of diabetes and is a major cause of new blindness in the working-age population of developed countries. While the exact pathogenic basis of this condition remains ill defined, it is clear that hyperglycaemia is a critical factor in its aetiology. Protein kinase C (PKC) activation is one of the sequelae of hyperglycaemia and it is thought to play an important role in the development of diabetic complications. This review questions the currently held dogma that PKC stimulation in diabetes is solely mediated through the overproduction of palmitate and oleate enriched diacylglycerols. Blood glucose concentrations are closely tracked by changes in the levels of free fatty acids and these, in addition to oxidative stress, may account for the aberrant activation of PKCs in diabetes. Little is known about why PKCs fail to downregulate in diabetes and efforts should be directed towards acquiring such information. Considerable evidence implicates the PKCbeta isoform in the pathogenesis of diabetic retinopathy, but other isoforms may also be of relevance. In addition to PKCs, it is evident that novel diacyglycerol-activated non-kinase receptors could also play a role in the development of diabetic complications. Therapeutic agents have been developed to inhibit specific PKC isoforms and PKCbeta antagonists are currently undergoing clinical trials to test their toxicity and efficacy in suppressing diabetic complications. The likely impact of these drugs in the treatment of diabetic patients is considered. PMID- 14737744 TI - All-cause mortality in diabetic patients treated with combinations of sulfonylureas and biguanides. AB - BACKGROUND AND AIMS: Increased mortality in diabetic patients treated with combinations of high doses of sulfonylureas and metformin was recently reported. This study was aimed at the assessment of mortality in patients treated with low dose combinations of sulfonylureas and biguanides. METHODS: An observational cohort study was performed on a consecutive series of 927 outpatients with type 2 diabetes mellitus. Of these patients, 376 were treated with combinations of sulfonylureas (mean daily dose +/- SD: 7.3 +/- 3.2 mg for glibenclamide, and 338 +/- 79 mg for chlorpropamide) and biguanides (daily dose: 1.1 +/- 0.3 g for metformin, 60.1 +/- 19.6 mg for phenformin). Mortality was assessed through a search in the City of Florence Registry Office, with an average follow-up of 55.1 months. RESULTS: After adjusting for other potential confounders (including age, duration of diabetes, BMI, hypertension, lipid profile, HbA1c, and insulin treatment), mortality was significantly higher in patients treated with combinations of sulfonylureas and biguanides than in the rest of the sample, (relative risk, RR: 2.08; 95% confidence interval, CI 1.18-3.67, and RR: 1.68; 95%CI 1.01-2.79 among women and men, respectively). CONCLUSIONS: A higher mortality was observed in patients treated with combinations of sulfonylureas and biguanides, even at low doses. Safety of such combinations deserves further investigation. PMID- 14737745 TI - Soluble adhesion molecules in Finnish schoolchildren with signs of preclinical type 1 diabetes. AB - BACKGROUND: We measured the circulating concentrations of the soluble forms of intercellular adhesion molecule-1, ICAM-1 (sCD54) and L-selectin (sCD62L) in 104 non-diabetic Finnish schoolchildren testing positive for one or more diabetes associated autoantibodies and in 104 autoantibody-negative children to elucidate the relationship between soluble adhesion molecules and humoral, genetic and metabolic markers of preclinical type 1 diabetes. METHODS: Specific enzyme-linked immunosorbent assays were used to analyse serum sICAM-1 and sL-selectin concentrations. RESULTS: The sICAM-1 and sL-selectin levels were comparable in the autoantibody-positive and control children, even when comparing children with multiple autoantibodies with those having one or no autoantibodies. The IA-2A titres in children testing positive for this autoantibody correlated with the sICAM-1 concentrations (rs=0.62, P=0.05), but otherwise no significant associations were seen between the autoantibody specificities and the concentrations of soluble adhesion molecules. Control children with HLA DQB1 genotypes conferring a low or decreased risk of type 1 diabetes had higher levels of sL-selectin than those with high or moderate risk genotypes (P=0.04). sL selectin concentrations were significantly increased in the autoantibody-positive children with a first-phase insulin response (FPIR) below the 5th (n=11;P=0.026) or 10th percentiles (n=17;P=0.009) relative to the children with a normal FPIR. No associations were observed between sICAM-1 concentrations and DQB1 genotypes or FPIR. CONCLUSIONS: The data indicate that there are a few conspicuous signs of endothelial/leukocyte activation reflected in increased circulating levels of soluble adhesion molecules in schoolchildren who are positive for markers of preclinical type 1 diabetes. The correlation between sICAM-1 concentrations and IA-2A levels in the IA-2A-positive children suggests that the former may increase in late preclinical type 1 diabetes, as IA-2A are the last autoantibodies to appear in the prediabetic process. Increased sL-selectin concentrations in subjects with impaired beta-cell function may reflect an active destructive insulitis process. PMID- 14737746 TI - Pramlintide reduces postprandial glucose excursions when added to insulin lispro in subjects with type 2 diabetes: a dose-timing study. AB - BACKGROUND: To assess the postprandial glucose-lowering effect of the human amylin analog pramlintide when given with insulin lispro in subjects with type 2 diabetes, with an emphasis on the optimal dose timing relative to meals. METHODS: In this randomized, single-blind, placebo-controlled, five-way crossover study, 19 subjects with type 2 diabetes using insulin lispro underwent five consecutive mixed-meal tests. In randomized order, subjects received subcutaneous injections of placebo at -15 min or 120-microg pramlintide at -15, 0, +15, or +30 min relative to the standardized breakfast after an overnight fast. Insulin lispro was injected at 0 min at doses that were adjusted appropriately for both the content of the standardized meal and the anticipated effects of pramlintide. Plasma glucose concentrations were measured before and during the 4-h postmeal period. RESULTS: When injected at 0 min, pramlintide reduced the postprandial glucose excursion by 81% compared to insulin lispro + placebo (incremental AUC(0 4 h) (mean +/- SE) 2.0 +/- 1.5 vs. 10.4 +/- 2.2 mmol/h/L, P<0.05). When pramlintide was injected at -15, +15, and +30 min, the postprandial incremental glucose AUC(0-4 h) was also significantly reduced (P<0.05), but to a lesser extent (42 to 73%). Pramlintide treatment was well tolerated and no serious adverse events were reported. CONCLUSIONS: Administration of pramlintide either at or just prior to a meal caused a greater reduction in postprandial glucose than either administration of placebo or postmeal pramlintide injections in subjects with type 2 diabetes treated with a rapid-acting insulin analog, insulin lispro. PMID- 14737747 TI - The role of the sodium hydrogen exchanger-1 in mediating diabetes-induced changes in the retina. AB - BACKGROUND: The sodium hydrogen exchanger (NHE) is a transmembrane protein responsible for alkalinization and control of intracellular acidosis by the removal of hydrogen and the subsequent influx of sodium. Our investigation attempts to determine the role of NHE-1 in the pathogenesis of early retinal microangiopathy due to diabetes. METHODS: Diabetes was induced in male Sprague Dawley rats with a single intravenous streptozotocin injection (65 mg/kg). To examine the duration-dependent changes in NHE-1 expression, retinas from 1-, 6- and 12-week diabetic animals were analyzed. To examine the functional consequences of NHE-1 inhibition in comparison with good blood glucose control, diabetic rats were randomly assigned to poorly controlled diabetic, well controlled diabetic, poorly controlled diabetic with cariporide groups and were compared with nondiabetic controls after six weeks. Cariporide is an orally active inhibitor of NHE-1 (6000 ppm in rat chow). At the end of the treatment period, color Doppler ultrasound was used to determine the resistivity index (RI) of the central retinal artery. The mRNA expression of endothelin (ET) isoforms 1 and 3, inducible and endothelial nitric oxide synthase (iNOS and eNOS respectively) and NHE-1 were examined. NHE-1 distribution was localized with immunohistochemistry. RESULTS: All diabetic animals showed hyperglycemia, increased glycated hemoglobin and lower body weight gain compared to nondiabetic controls. Diabetes caused an increased RI, indicative of retinal vasoconstriction, which was corrected by both cariporide treatment and good glucose control. NHE-1 was localized in the endothelium of the retinal microvasculature and the neuronal and glial components. NHE-1 mRNA expression was unchanged after 1 week and increased after 6 and 12 weeks of diabetes. Furthermore, a diabetes-induced upregulation of ET-1 and ET-3 mRNA expression after six weeks was corrected with cariporide treatment. NHE-1 inhibition of diabetic animals upregulated iNOS mRNA levels, although expression of eNOS and iNOS mRNA were not altered in poorly controlled diabetes. Improved blood glucose control with higher doses of insulin also corrected diabetes-induced increased RI by upregulating eNOS and iNOS mRNA expression. CONCLUSIONS: The results of the study suggest that NHE-1 may be involved in the regulation of several vasoactive modulators that contribute to functional alterations in diabetic retinal microangiopathy. PMID- 14737748 TI - Glutathione metabolism and oxidative stress in neonatal rat tissues from streptozotocin-induced diabetic mothers. AB - BACKGROUND: Diabetes in pregnancy has been reported to cause congenital malformations in the offspring. Hyperglycemia-induced teratogenicity is related to the increased oxidative stress and depletion of glutathione (GSH) in fetal tissues. METHODS: In the present study, we have examined the levels of reactive oxygen species (ROS) and peroxide production in the tissues of neonatal rats and in the pancreas of diabetic mother rats. Diabetes was induced one day after pregnancy by a single injection of streptozotocin (60 mg/kg body weight). Different tissues from one-day-old neonates and the pancreas from the mothers were collected to study the level of oxidative stress and GSH metabolism. RESULTS: An increase in ROS and peroxide production was observed in the pancreas of diabetic rats and in the liver, kidney, brain and skin of the neonates of the diabetic mothers. The catalytic activity of cytochrome P450 2E1 (CYP2E1) was also increased in these tissues from diabetic rats. With the exception of the kidney, the GSH concentration was significantly lower in the tissues of neonates and in the pancreas of diabetic mothers. CONCLUSION: This reduction in GSH concentration was presumably associated with its increased utilization, due to increased production of peroxides and also due to its reduced regeneration. The study may have an implication in understanding the pathology of diabetic complications in pregnancy and sensitivity toward antioxidant therapy during pregnancy. PMID- 14737749 TI - Current literature in diabetes. PMID- 14737750 TI - Using simulation-based inference with panel data in health economics. AB - Panel datasets provide a rich source of information for health economists, offering the scope to control for individual heterogeneity and to model the dynamics of individual behaviour. However the qualitative or categorical measures of outcome often used in health economics create special problems for estimating econometric models. Allowing a flexible specification of the autocorrelation induced by individual heterogeneity leads to models involving higher order integrals that cannot be handled by conventional numerical methods. The dramatic growth in computing power over recent years has been accompanied by the development of simulation-based estimators that solve this problem. This review uses binary choice models to show what can be done with conventional methods and how the range of models can be expanded by using simulation methods. Practical applications of the methods are illustrated using data on health from the British Household Panel Survey (BHPS). PMID- 14737751 TI - The effects of price and policy on marijuana use: what can be learned from the Australian experience? AB - This research examines the responsiveness of the demand for marijuana to changes in its money price and criminal status using data on individuals from the Australian National Drug Strategy's Household Surveys (NDSHS). The results suggest that both the prevalence of marijuana use and the conditional demand for marijuana in the general population are responsive to changes in its money price. Significant differences are found in the effect of price on participation in marijuana use across age-groups, with participation by youth more price sensitive than participation by older age-groups. Similarly, the effect of the legal status of marijuana use on the participation decision is found to differ across age groups and gender. Specifically, decriminalisation is associated with an increases in the prevalence of use by males over the age of 25. There is no evidence that decriminalisation significantly increases participation in marijuana use by either young males or females, or that decriminalisation increases the frequency of use among marijuana users. PMID- 14737752 TI - Does problem drinking affect employment? Evidence from England. AB - In this paper, we use data from the Health Survey of England to show that problem drinking is negatively and significantly associated with the probability of being in work, once the endogenous relationship between these outcomes is accounted for. Being a problem drinker leads to a substantial reduction in the probability of working by between 7 and 31%, the former figure being roughly equivalent to the positive effect of having a degree relative to no qualifications in our data. This finding is robust to a variety of identifying restrictions and definitions of problem drinking. Moreover, we find that problem drinking defined by the observed psychological and physical symptoms of alcohol is an important predictor of employment, and allows for the fact that individuals differ in their tolerance or susceptibility to alcohol. Our results suggest that there may be important labour market benefits from public health policies aimed at the prevention and treatment of problem drinking. PMID- 14737753 TI - Excess capacity and expense preference behaviour in National Health Systems: an application to the Spanish public hospitals. AB - The aim of this paper is to study excess capacity in National Health System (NHS) hospitals, which have the characteristics of a bureaucracy, within a theoretically consistent framework. In this sense, we develop and estimate an empirical model which has not previously been applied to this problem. This model, based on an input distance function, is attractive for at least two reasons. Unlike the production function, the input distance function is valid for multiproduct technologies such as that of hospitals, and in contrast to the cost function it permits testing of the cost minimisation hypothesis and therefore allows expense preference behaviour in the context of NHS hospitals. We present an application to the Spanish public hospital sector and observe persistent allocative inefficiency in variable inputs and overcapitalisation in these hospitals. Hence, our results suggest that this sector is not in long-run equilibrium and that adjustments in variable inputs and capital equipment investments are necessary to control hospital cost. PMID- 14737754 TI - Lack of multiplicative transitivity in person trade-off responses. AB - BACKGROUND: The person trade-off (PTO) is a technique for eliciting preferences for resource allocation across patient groups. In principle PTO responses should satisfy a requirement of multiplicative transitivity, i.e. that if people consider treatment of 1 in state A to be equivalent to treating 10 in state B, and 1 in state B to be equivalent to 10 in state C, then they should find 1 in state A equivalent to 100 in state C. Earlier studies addressing labelled diseases (specific diagnoses), have shown multiplicative intransitivity of the PTO responses. Our purpose was to test multiplicative transitivity in the case of health states described with the EuroQol instrument only and to find a possible framing effect such as the number of persons in the reference intervention. METHODS: Forty-four master degree students were asked to fill in a questionnaire addressing four chronic health states. Their task consisted in (1). ranking the states by severity, (2). valuing each of them by the means of the time trade-off, and (3). doing the PTO for all the 10 possible pairwise combinations of the four chronic states plus a fatal one. In a subsequent questionnaire the number of persons in the reference intervention in the PTO was increased from 10 to 100. Multiplicative transitivity was studied in subjects who demonstrated a willingness to trade off and consistency in ranking individual values. RESULTS: None of the 39 subjects included satisfied a minimum multiplicative transitivity requirement in PTO responses. Internal consistency was not improved when the PTO involved health states close to each other in terms of severity, nor when the prevention of death was not the reference intervention. For the 22 subjects having answered both types of questionnaire, increasing the number of persons in the reference intervention did not improve multiplicative transitivity. CONCLUSIONS: The PTO holds promise as a useful method for determining social preferences for priority setting, inasmuch as it captures distributive concerns that individual utility techniques such as the time trade-off do not address. But the lack of multiplicative transitivity in PTO responses is unsatisfactory, and ways to reduce this problem need to be explored. PMID- 14737755 TI - Evidence of range bias in contingent valuation payment scales. AB - The payment scale format has been widely used in willingness-to-pay studies in health care. Concerns have been expressed that the format is, in theory, prone to range bias, although this proposition has not been tested directly. We report the findings of a contingent valuation questionnaire study of colorectal cancer screening, wherein different subjects were provided with payment scales of two different lengths. Whilst the long-scale instrument included scale values up to pound 1000, the short-scale version extended only to pound 100. After controlling for inter-sample differences in, for example, income, education and health behaviour, it emerged that the long-scale instrument produced a mean willingness to pay more than 30% higher than that resulting from the short-scale version. We believe our findings to be strongly supportive of the likelihood of range bias in payment-scale instruments, with important consequences for the estimation of both average valuation and consumer surplus. PMID- 14737756 TI - Bayesian estimation of cost-effectiveness: an importance-sampling approach. AB - We describe a method for estimating the cost-effectiveness of a new treatment compared to a standard, using data from a comparative clinical trial. We quantify the clinical effectiveness as a binary variable indicating success or failure. The underlying statistical model assumes that costs are uncensored and follow separate gamma distributions in each of the groups defined by the four possible combinations of treatment arm and effectiveness outcome. The method is subjectivist, in that it represents prior uncertainty about model parameters with a probability distribution, which we update via Bayes's theorem to produce a posterior distribution. We approximate the posterior by importance sampling, a straightforward simulation method. We illustrate the method with an analysis of cost (derived from resource usage data) and effectiveness (measured by one-year survival) in a clinical trial in heart disease. The example demonstrates that the method is practical and provides for a flexible data analysis. PMID- 14737757 TI - Managed care and shadow price. AB - A managed-care company must decide on allocating resources of many services to many groups of enrollees. The profit-maximizing allocation rule is characterized. For each group, the marginal utilities across all services are equalized. The equilibrium has an enrollee group shadow price interpretation. The equilibrium spending allocation can be implemented by letting utilitarian physicians decide on service spending on an enrollee group subject to a budget for the group. PMID- 14737758 TI - Formation of defined microporous 3D structures starting from cross-linked hydrogels. AB - A new and simple technique was developed to obtain polysaccharide (hyaluronane, alginate and carboxymethylcellulose) -based hydrogels with a defined porous morphology. The technique consists of stratifying a cross-linked hydrogel on a filter with known pore diameter. CO(2) bubbles, produced by the addition of HCl to a porogen salt NaHCO(3), are forced to pass through the filter, and they induce the hydrogel to assume a porous morphology. The presence and distribution of pores was confirmed by scanning-electron microscopy (SEM). A strict correspondence was found between the porosity of the filter and the pore diameter in the hydrogels. Water uptake measurements showed a decreased amount of water taken up by the porous hydrogels compared with the native hydrogels, due to a compacting of the material. An explanation of the porous material properties of Hyal hydrogel was given on the basis of FTIR spectra. PMID- 14737759 TI - Bone regeneration in rabbit sinus lifting associated with bovine BMP. AB - Autogenous bone is considered the optimal grafting material for sinus lifting, although its harvesting causes great patient discomfort. Various approaches have been taken in order to obtain sinus lifting with preexisting tissue. However, because of the unsuitability of such tissue, additional materials have been required. Alternatively, biomaterials from humans or other animals are used. In this study, the efficacy of using morphogenetic bovine bone protein (BMPb) to augment the maxillary sinus floor was examined. Four grafting materials were employed: lyophilized bovine bone powder, absorbable collagen flakes, natural hydroxylapatite, and synthetic hydroxylapatite. Two groups of rabbits were studied. In one group, graft material only was used. In the other, graft material was combined with 0.5 mg BMPb. During 8 weeks of observation, polyfluorochrome tracers were injected in subcutaneous tissue to evaluate new bone- deposition periods. Following sacrifice, the samples were examined under fluorescent and light microscopes. Results indicated 33.34% more newly formed bone in BMPb animals than in controls. Graft-material resorption increased, but natural HA showed no significant alterations. The results show that the use of BMPb, although providing osteoinduction, might not promote sufficient bone formation. Nonetheless, this material could provide an alternative to autogenous grafts, thereby avoiding patient discomfort. PMID- 14737760 TI - In vivo aging test for a bioactive bone cement consisting of glass bead filler and PMMA matrix. AB - The degradation of a new bioactive bone cement (GBC), comprised of an inorganic filler (bioactive MgO-CaO-SiO(2)-P(2)O(5)-CaF(2) glass beads) and an organic matrix [high-molecular-weight polymethyl methacrylate (PMMA)], was evaluated in an in vivo aging test. Hardened rectangular specimens (20 x 4 x 3 mm) were prepared from two GBC formulations (containing 50% w/w [GBC50] or 60% w/w [GBC60] bioactive beads) and a conventional PMMA bone cement control (CMW-1). Initial bending strengths were measured with the use of the three-point bending method. Specimens of all three cements were then implanted into the dorsal subcutaneous tissue of rats, removed after 3, 6, or 12 months, and tested for bending strength. The bending strengths (MPa) of GBC50 at baseline (0 months), 3, 6, and 12 months were 136 +/- 1, 119 +/- 3, 106 +/- 5 and 104 +/- 5, respectively. Corresponding values were 138 +/- 3, 120 +/- 3, 110 +/- 2 and 109 +/- 5 for GBC60, and 106 +/- 5, 97 +/- 5, 92 +/- 4 and 88 +/- 4 for CMW-1. Although the bending strengths of all three cements decreased significantly from 0 to 6 months, those of GBC50 and GBC60 did not change significantly thereafter, whereas that of CMW-1 declined significantly between 6 and 12 months. Thus, degradation of GBC50 and GBC60 does not appear to continue after 6 months, whereas CMW-1 degrades progressively over 12 months. Moreover, the bending strengths of GBC50 and GBC60 (especially GBC60) were significantly higher than that of CMW-1 throughout. It is believed that GBC60 is strong enough for use under weight bearing conditions and that its mechanical strength is retained in vivo; however, its dynamic fatigue behavior will need assessment before application in the clinical setting. PMID- 14737761 TI - Migration of polyethylene debris along well-fixed cemented implants. AB - Implants, consisting of smooth Inox cylinders, were cemented into the lower femur and upper tibia of nine sheep to study the distal migration of polyethylene particles. Some implants had a titanium-bead porous coat at the proximal end. These were of three types: In the first type, the porous coat was covered with hydroxyapatite to obtain a bony seal; the second type was prepared for a polymethylmethacrylate seal; in the third type, the porous zone was surrounded by a 2-mm-thick space to allow the formation of a fibrous seal. Small polyethylene particles were injected into the knees once a week during the third and fourth months after implantation. The animals were euthanized 2 months later. Major longitudinal sections of the implants and the surrounding bone were examined under a polarized light microscope. Birefringent particles were counted at the cement-bone and cement-implant interfaces. Osteolysis was not observed. None of the seals significantly decreased the migration of particles around the cemented part of the implants. Particles were observed in cement fissures and vacuoles. They migrated at both interfaces and in the bone itself. They were visible in marrow spaces between bone trabeculae. PMID- 14737762 TI - Biological properties of acid etched titanium implants: effect of sandblasting on bone anchorage. AB - The SLA (sandblasted with large grit and acid etched) surface is a textured surface that has been documented to lead to a rapid and strong implant fixation. The purpose of the present study was to determine the contribution of sandblasting in addition to etching to implant anchorage. It was also aimed to determine if the pits carved during etching alone have a bone-interlocking capacity that leads to microanchorage between the implant and bone. SLA implants and machined-and-acid-etched (MA) implants were placed in the maxilla of Land Race pigs. After 10 weeks of healing, they were reverse torqued. The reverse torque of the SLA and MA implants was 157.29 +/- 38.04 N cm and 105.33 +/- 25.12 N cm, respectively. Sandblasting increased bone anchorage by 49.3%; the difference was statistically significant (p =.028). Bone was found attached to both surfaces; bone ingrowth was found in the pits of both surfaces. It is suggested that the two surfaces are able to generate bone interlocking and mechanical coupling at the interface. When finite-element modeling is performed with these surfaces, it is suggested that the bound mode be used instead of the slip mode. PMID- 14737763 TI - Electric-current-induced detachment of Staphylococcus epidermidis strains from surgical stainless steel. AB - Infection of percutaneous biomaterials implants, such as fixation frames used for the repair of complicated fractures in orthopedics, is a major complication that almost inevitably leads to replacement of the implant. As antibiotic therapy usually has little impact on biomaterial-associated infections, it is the aim of this article to examine whether implant-associated Staphylococcus epidermidis and Staphylococcus aureus strains could be stimulated to detach from a surgical stainless steel anode during application of an electric current. First, bacteria were allowed to adhere from a flowing suspension of physiological ionic strength in a parallel plate flow chamber to a stainless-steel surface, after which the suspension was replaced by a bacterium-free solution with a specified ionic strength (0.5-150-mM potassium phosphate). DC currents ranging from 15 to 125 microA were applied to induce bacterial detachment. Initial detachment decreased with increasing ionic strength at 100 microA. The percentage detachment achieved by application of an electric current after 2.5 h was highest (95%) in 1-mM potassium phosphate and decreased to 15% when the ionic strength exceeded 40 mM. The electric current did not significantly affect the percentage detachment, but initial detachment rates increased with increasing current from 1000 cm(-2) s(-1) at 15 microA to 7000 cm(-2) s(-1) at 125 microA. Although different isolates of S. epidermidis and S. aureus showed different patterns of current-induced detachment, all strains could be stimulated to detach. The results of this study define ionic-strength conditions and electric currents yielding staphylococcal detachment from surgical stainless steel and therewith point to a pathway for the treatment and prevention of percutaneous metal-implant infection. PMID- 14737764 TI - SEM/EDS evaluation of porcelain adherence to gold-coated cast titanium. AB - The adhesion between titanium and dental porcelain is related to the diffusion of oxygen to the reaction layer formed on cast-titanium surfaces during porcelain firing. The diffusion of oxygen could be suppressed by coating the titanium surface with a thin gold layer. This study characterized the effects of gold coating on titanium-ceramic adhesion. ASTM grade II CP titanium was cast into a MgO-based investment (Selevest CB, Selec). The specimen surfaces were air abraded with 110-microm Al(2)O(3) particles. Gold coating was applied on titanium surfaces by three methods: gold-paste (Deck Gold NF, Degussa-Ney) coating and firing at 800 degrees C for three times, single gold-paste coating and firing followed by sputter coating (40 mA, 500 s), and sputter coating (40 mA, 1000 s). Surfaces only air abraded with Al(2)O(3) particles were used as controls. An ultra-low-fusing dental porcelain (Vita Titankeramik, Vident) was fused on titanium surfaces. Specimen surfaces were characterized by SEM/EDS and XRD. The titanium-ceramic adhesion was evaluated by a biaxial flexure test (N = 8), and area fraction of adherent porcelain (AFAP) was determined by EDS. Numerical results were statistically analyzed by one-way ANOVA and the Student-Newman-Keuls test at alpha = 0.05. SEM fractography showed a substantial amount of porcelains remaining on the gold-sputter-coated titanium surfaces. A new Au(2)Ti phase was found on gold-coated titanium surface after the firing. Significantly higher (p <.05) AFAP values were determined for the gold-sputter-coated specimens compared to the others. No significant differences were found among the other groups and the control. Results suggested that gold coatings used in this study are not effective barriers to completely protect titanium from oxidation during the porcelain firing, and porcelain adherence to cast titanium can be improved by gold-sputter coating used in the present study. PMID- 14737765 TI - Effect of post-processing heat treatment on the fracture strength of a heat pressed dental ceramic. AB - The purpose of this study was to evaluate the effect of post-processing heat treatment on the fracture strength of a heat-pressed lithium disilicate dental ceramic (OPC 3G). Bar-shaped wax patterns (2 x 4 x 22 mm; n = 60) were invested and heat pressed in an automated pressing machine according to manufacturer's recommendations. The bars were polished to 1200 grit and annealed at 600 degrees C for 1 h. Two groups were left untreated as controls, one of which was Vickers indented in oil prior to testing in three-point bending. Two groups were indented and then heat treated at either 800 or 825 degrees C for 30 min. The corresponding control groups were heat treated, then indented prior to testing. Optical micrographs of the indentations were taken before and after heat treatment under differential interference contrast. The fracture toughness and/or fracture strength of the specimens was determined by the indentation-strength technique in three-point bending. Indentation-generated cracks were not discernible by optical microscopy after heat treatment at either 800 or 825 degrees C for 30 min, even under 1000 x magnification. ANOVA and Tukey's test showed that heat treatment in air at 825 degrees C for 30 min significantly increased the fracture strength of a heat-pressable lithium disilicate ceramic. This result was attributed to healing of the flaws present in the material. PMID- 14737766 TI - The development of theoretical relationships between some handling parameters (setting time and setting temperature), composition (relative amounts of initiator and activator) and ambient temperature for acrylic bone cement. AB - Commercially available acrylic bone cements are two-component systems based on the polymerization of methyl methacrylate around poly(methyl methacrylate) particles. When benzoyl peroxide (BPO), which is the initiator, in the powder component meets accelerator (N,N-dimethyl-p-toluidine (DMpT)) in the liquid component, radicals are produced, initiating the polymerization. This solidifies the cement. In this work, kinetic expressions have been developed that describe the relationship between bone cement setting time on the one hand, and BPO and DMpT concentrations on the other. Changes in setting time with ambient temperature follow a complex relationship, because both the polymerization process (initiation, propagation, and termination) and the swelling and dissolution of the polymer particles contribute to setting. The contribution of polymer swelling and dissolution to the setting process was determined by developing a relationship between the doughing time, which is substantially independent of DMpT or BPO concentrations, and ambient temperature. A value of 64 kJ mol(-1) was found for the activation energy for this process. An activation energy for the overall setting process of 68 kJ mol(-1) was determined from setting-time measurements over several ambient temperatures. This indicates that the sensitivity of setting time to temperature depends more on swelling and dissolution than on the polymerization process. PMID- 14737767 TI - Effects of the initial temperature of acrylic bone cement liquid monomer on the properties of the stem-cement interface and cement polymerization. AB - It has been shown that preheating the femoral stem prior to insertion minimizes interfacial porosity at the stem-cement interface. In this study, the effects of methylmethacrylate monomer temperature prior to mixing on the properties of stem cement interface and cement polymerization were evaluated for 4 degrees C, room temperature, and 37 degrees C using a test model and cementing techniques that simulated a clinical situation. The nature and extent of interfacial porosity of stem-cement interface was quantified, the static shear strength of the stem cement interface determined, and the time and temperature of polymerization at the cement-bone interface were measured. Compared to RT monomer, preheating monomer to 37 degrees C produced higher polymerization temperatures and greater initial interfacial shear strength with an unchanged amount of interfacial porosity. Precooling monomer to 4 degrees C produced lower polymerization temperatures and decreased initial interfacial shear strength, with the amount of interfacial porosity unchanged compared to the RT group. Although clinical techniques of preheating or precooling bone cement have some effects on the properties of the stem-cement interface and cement polymerization, they do not appear to enhance implant fixation. PMID- 14737768 TI - Segmented detachable structure of cochlear-implant electrodes for close-hugging engagement with the modiolus. AB - Surface and bulk modification techniques of polydimethylsiloxane (PDMS) polymers were used to develop a new intracochlear electrode that can closely hug the inner wall of scala tympani. Laser-induced surface grafting of poly (2-hydroxyethyl methacrylate) (PHEMA) and sequential method for preparation of interpenetrating polymer networks (IPNs) of PDMS/PHEMA were, respectively, used for surface and bulk modifications. The hydrogel content and water-uptake capability of the modified samples were optimized by Taguchi method for experimental design. The modified PDMS samples were examined by performing attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy, water contact-angle measurement, and peel strength tests. The performances of actual-sized fabricated electrodes were assessed inside a transparent model of scala tympani, which was filled with saline. After a swelling process, the hydrophilic branch begins to elongate and detach from hydrophobic branch and pushes it toward the inner wall. PMID- 14737769 TI - Bone-defect healing with calcium-sulfate particles and cement: an experimental study in rabbit. AB - Calcium sulfate (CaS) has been shown to be a reasonable alternative to autogenous bone graft for treating bone lesions in dentistry. The aim of this work was an histological study of the bone healing of defects treated with calcium sulfate in the form of cement or beads, in animal. Eight New Zealand rabbits, weighing about 2.5 Kg were used in this study. In each rabbit, four 6 mm bone defects were created in the tibial metaphysis. The 2 defects in the right tibia were filled with calcium sulfate as cement, while the 2 defects in the left one were filled with calcium sulfate as beads. Four rabbits were killed after respectively 2 and 4 weeks, with an intravenous injection of Tanax, and the block sections, containing the bone defects, were retrieved. A total of 16 defects filled by cement and a total of 16 defects filled by beads were retrieved. The specimens were processed to obtain thin ground sections with the Precise 1 Automated System. In the first phases of healing it was possible to observe an intense osteoblastic activity, and in some areas osteoid matrix was present. After two weeks the calcium sulfate (both cement and beads) was still present, and biological fluids and cells were present inside the material. Newly formed bone surrounded the calcium sulfate and filled about 10% of the defect. After four weeks the calcium sulfate was almost completely resorbed and substituted by new bone. Approximately 34% of the defects were filled by newly formed bone. BEI and XRM evaluations showed the structural components of the filled defects. In none of the specimens were inflammatory cells present. No significant differences were found using both calcium sulfate as cement and beads, and they both have shown a high biocompatibility, appearing to promote newly bone formation in the rabbit model, and they did not induce any untoward effect on the bone regeneration processes. PMID- 14737770 TI - Properties and biocompatibility of polypropylene graft copolymer films. AB - Modifying the surfaces of polymers has received a great deal of attention, because it could bring about specific surface characteristics such as antithrombogenic property. Therefore, N-vinyl pyrrolidone/sodium acrylate (NVP/Na AAc) binary monomers were introduced onto polypropylene (PP) films by a radiation grafting method. The effect of solvent and comonomer composition on the degree of grafting was determined. Studies of the mechanical properties and water content of such graft copolymers showed that as the grafting yield increases the elongation percent decreases. However, the water content increases with increasing grafting yield. The blood compatibility of the original PP and PP-g NVP/Na-AAc films was evaluated by determination of the extent of platelet adsorption and thrombus formation. The blood compatibility of PP-g-NVP/Na-AAc seems to be better than that of original PP. PMID- 14737771 TI - Change of properties during storage of a UDMA/TEGDMA dental resin. AB - The aim of this study was to evaluate the changes in viscoelastic properties of a UDMA-based dental resin as a function of time after initial light exposure. Specimens of a UDMA/TEGDMA (70:30 wt%) resin were irradiated by a visible-light curing unit. Immediately after the irradiation, the light-cured specimen was stored in the dark for different times from 1 to 120 h at 37 degrees C, and characterized by means of DMA, DSC, and FTIR spectroscopy. The irradiated specimen exhibited a bimodal shape in the form of two rapid declines in log E' corresponding to glass transition with a plateau between the two declines. Two distinct peaks were seen in tan delta versus temperature. The thermal reaction of the incompletely cured sample with residual groups trapped by the fast reaction during irradiation is responsible for the plateau. After storage, significant changes were observed in dynamic mechanical parameters, DSC exotherm, and degree of conversion. Storage modulus continued to increase during the 4 h of storage and leveled off thereafter. Peak heights of tan delta versus temperature were also influenced by storage. Degree of conversion increased from 75 +/- 2% immediately after irradiation to 87 +/- 3% after 120 h storage. The changes of the properties of this dental resin system when stored at 37 degrees C after irradiation are clinically important in terms of stability, durability, and performance after initial polymerization. PMID- 14737772 TI - The effect of tranexamic acid in fibrin sealant on adhesion formation in the rat. AB - The objective of the research was to determine the effect of the type, dose, and volume of anti-fibrinolytic agents (tranexamic acid, aprotinin) added to fibrin formulations, on adhesion development. Adhesions were induced in 228 male rats by creating apposing parietal and visceral peritoneal defects. Animals were randomized to receive no treatment or a fibrin formulation containing aprotinin or tranexamic acid. Seven days later the incidence of adhesions, and the force and energy required to detach them, were determined. Adhesions developed in 13/13 rats in the control and aprotinin groups. Treatment with fibrin (100 mg/ml tranexamic acid) resulted in adhesions in 4/14 rats (as strips, p < or = 0.0005), 4/10 rats (as spray, p < or = 0.0036), and 12/15 rats (by drip). The reduction of adhesions was dependent on the concentration of tranexamic acid with strip and spray application. Using commercial formulations, tranexamic-acid-containing fibrin (10/15, p = 0.042), but not aprotinin-containing fibrin (13/15), reduced the incidence of side-wall adhesions from 15/15 in controls. Fibrin containing either tranexamic or aprotinin reduced the incidence and severity of adhesions. This effect was greater when tranexamic acid was used and was dependent on the mode of administration, the volume, and to a degree, the concentration of tranexamic acid. PMID- 14737773 TI - Drs. Dobkin and Bernatsky reply. Publication of articles pertaining to fibromyalgia. PMID- 14737774 TI - [Transfusion and hemostasis: unavoidable commitment for modern anesthesiology]. PMID- 14737775 TI - [Update on transfusion practice among anesthesiologists and its impact on the surgical patient]. AB - OBJECTIVE: To determine the effectiveness of a new educational program on transfusion practice, directed to the staff of an anesthesia and postoperative recovery service, by evaluating its impact on intraoperative transfusion (IOT). MATERIAL AND METHODS: We reviewed the incidence of IOT during the first semesters of 1996 and 2001 for general, urologic, otolaryngologic, maxillofacial, thoracic, and vascular surgery. Other factors such as sex, age, type of intervention, emergency status, duration of operation, and use of blood products were also taken into consideration. RESULTS: A statistically significant overall reduction in IOT occurred between 1996 (4.9%) and 2001 (3.6%). The decrease in transfusions (a reduction of 18.8% in transfused patients) was even greater in general surgery and urology, particularly in scheduled surgery (4.1% in 1996 vs 2.6% in 2001). However, the total use of packed red blood cells did not change inasmuch as the number of units per patient was higher in 2001 (2.8 units/patient) than in 1996 (2.4 units/patient). In emergency surgery, the IOT rate increased from 7.6% in 1996 to 8.1% in 2001. We also noticed a higher rate of multiple transfusions (defined as the use of 5 or more units of packed red blood cells during surgery) in 2001. CONCLUSION: The introduction of an educational program directed to anesthesiologists has been useful for reducing IOT, although the overall use of blood products has not decreased. PMID- 14737776 TI - [Spinal cord compression caused by hematoma related to neuroaxial anesthesia in Spain]. AB - OBJECTIVES: Spinal cord compression from a hematoma is a rare serious complication of neuroaxial anesthesia. Our objective was to investigate cases reported and published by Spanish authors. MATERIAL AND METHODS: Cases of spinal cord compression related to neuroaxial hematomas (epidural, subarachnoid, or both) reported at congreses and meetings in Spain or published in Medline-indexed journals from 1989 through December 2002 were reviewed. The clinical characteristics, risk factors, treatments, and outcomes were described for each case. RESULTS: Since 1996, when the first cases were reported, 20 cases in all have appeared: 8 related to sub-arachnoid anesthesia, 8 to epidural anesthesia, 1 to a combination, and 1 to diagnostic lumbar puncture. In 2 cases, the neuroaxial technique used was poorly defined. Factors that might have caused the complication could be identified in 11 cases, among which there were 7 cases of anesthetic puncture or manipulation of an epidural catheter during a period of hemorrhagic risk because of antithrombotic therapy. The hematoma was evacuated surgically in 11 cases, and medical treatment was provided in 9 cases. The neurologic outcome was satisfactory in 14 cases. CONCLUSIONS: The number of compressive spinal hematomas reported or published by Spanish authors is fairly high, and there are cases related to both sub-arachnoid and epidural anesthesia. Nonsurgical treatment was provided in 45% of the cases and the outcome was satisfactory in 70%. Risk factors were identified in over half of the reported cases. PMID- 14737777 TI - [Peripheral nerve block for surgical anesthesia and postoperative analgesia of the legs]. AB - Even though peripheral nerve blocks (PNB) on the lower limb offer advantages over neuroaxial blockades and general anesthesia, their use has not been fully established. The infrequency with which PNBs are used may be due to inadequate learning, the need to make several injections, the time until onset of block, or anesthesiologists' and surgeons' lack of familiarity with the benefits of regional blocks. Interest seems to have increased in recent years, as numerous publications have focused on lower limb PNBs for surgery and postoperative analgesia. Our aim was to review the main approaches used and the indications for each. PMID- 14737778 TI - [Cardiac tamponade caused by aortic pseudoaneurysm with fistula in the right ventricle]. AB - A 69-year-old woman, with an aortic replacement valve for rheumatic aortic insufficiency suffered a sudden cardiac tamponade caused by a ruptured pseudoaneurysm of the ascending aorta which had its origin in the suture of the aorta performed 10 years earlier. The cardiac tamponade presented as syncope, absent central pulse, and respiratory failure. Hemodynamic variables recovered with early detection and start of cardiopulmonary resuscitation maneuvers. After fluids were replaced and following catecholamine treatment, a transesophageal echocardiogram revealed a pseudoaneurysm and continuous flow between the aorta and the right ventricle. The aneurysm and an unusual right ventricular fistula were repaired surgically through an emergency exploratory sternotomy. Cardiovascular recovery was satisfactory although hospital discharge was delayed because of pulmonary complications leading to severe hypoxia, pleural effusion, and pneumonia, with tracheal aspirate and bronchoalveolar lavage cultures positive for Pseudomonas aeruginosa, Acinetobacter spp, and Aspergillus flavus. We stress that immediate start of cardiopulmonary resuscitation prevented multiorgan failure and allowed for satisfactory recovery from surgery in spite of the presence of risk factors for mortality. PMID- 14737779 TI - [Treatment with desmopressin before epidural anesthesia in a patient with type I von Willebrand disease]. AB - A 33-year-old primipara with von Willebrand disease type I was admitted in labor at 37 weeks, requesting epidural analgesia. The consultant hematologist advised treating with desmopressin acetate (DDAVP) before inserting an epidural catheter. Desmopressin at a dose of 0.3 microgram/Kg was administered intravenously and the catheter was inserted to L3-L4 to infuse 0.1% bupivacaine with 2 micrograms/mL of fentanyl at a rate of 12 mL/h. Four hours later the patient was brought to the operating room for forceps delivery of a healthy boy. One hour later, she had recovered normal motor tone followed by normal sensitivity in the lower extremities. The catheter was then withdrawn with no signs of bleeding. A woman with von Willebrand's disease can receive an epidural block for analgesia during childbirth. The decision to perform the block should be individualized, based on coagulation tests. DDAVP may play a role in improving hemostasis. PMID- 14737780 TI - [Peripartum cardiomyopathy and preeclampsia: an association with fatal outcome]. AB - A 33-year-old obese gypsy woman was admitted in labor with signs of preeclampsia followed by sudden pulmonary edema and cardiac arrest. Cesarean delivery was performed during resuscitation maneuvers, but both mother and fetus died. The diagnosis of peripartum cardiomyopathy was confirmed by pathology. The association of latent peripartum cardiomyopathy and episodes of hypertension greatly increases the risk of maternal and fetal death. Careful management of anesthesia and rigorous measures are necessary with a view to possible litigation. PMID- 14737781 TI - [A case of "ventilation difficulty-intubation difficulty" solved with transtracheal jet ventilation]. AB - A 56-year-old man recovering from a glossectomy and radical neck dissection presented severe oral bleeding, tracheal deviation with an asphyxiating hematoma and cyanosis. When 2 attempts at orotracheal intubation with the patient awake failed, transtracheal jet ventilation was used temporarily until a definitive airway could be established. Transtracheal jet ventilation is highly useful for managing an airway and maintaining gas exchange in life-threatening situations in which intubation and ventilation has become impossible, yet it is rarely used for that purpose. An easy, fast procedure that has not been widely used in Spain, this technique provides effective ventilation and oxygen while a definitive resolution of the emergency is sought. PMID- 14737782 TI - [Two cases of severe sepsis successfully treated with activated protein C]. AB - A 64-year-old man and a 52-year-old woman in shock with multiple organ failure and worsening of sepsis related organ failure assessment SOFA scores in the early days of care were treated with recombinant human activated protein C (rhAPC). In the woman sepsis was associated with reversible heart failure, with decreased ejection fraction, biventricular dilatation, and a sharp increase of troponin I, observations that have been linked to a higher rate of multiorgan failure and higher mortality. The man began to improve after 24 hours and the woman after 48 hours of rhAPC treatment, with both continuing to improve after withdrawal of treatment. Severe sepsis remains a therapeutic challenge. Among the many treatments based on the physiopathology of the disease, so far only rhAPC seems to improve outcome and reduce mortality. PMID- 14737783 TI - [Central nervous system complication of spinal surgery in prone position]. PMID- 14737784 TI - [Fissure of the azygos vein. Infrequent complication of chamber catheter]. PMID- 14737785 TI - [Anesthesia in nasofacial Wegener's disease]. PMID- 14737786 TI - [Intraoperative thyrotoxicosis in a female patient without diagnosed hyperthyroidism undergoing abdominal surgery]. PMID- 14737787 TI - [Special precautions for anesthesia in women reproductive age]. PMID- 14737788 TI - [Systemic monitoring versus multimodal neuromonitoring]. PMID- 14737789 TI - [In memory of Dr. Jesus Sahagun de la Lastra, one of the pioneers of obstetric analgesia-anesthesia in our country]. PMID- 14737790 TI - Case report: management of the flabby ridge: re-visiting the principles of complete denture construction. AB - Displaceable tissue on edentulous ridges may present a considerable clinical challenge to dental practitioners when providing complete dentures. Surgical excision techniques or use of dental implants has provided clinicians with methods of addressing this particular difficulty. However careful consideration and application of the principles of complete denture construction can provide a less invasive and less complex form of treatment. This article describes one such clinical scenario, and demonstrates the use of a suitable impression technique that is based on successful manipulation of the thermoplastic nature of the impression material. PMID- 14737791 TI - Influence on dimensional accuracy of volume of wash material introduced into pre spaced putty/wash impressions. AB - This study compared the accuracy of stone models obtained from two-stage, pre spaced putty/wash impressions under conditions in which known volumes of wash material were introduced during the second stage of the impression: Group I, a quantity of wash material corresponding to the space provided; Group II, double the quantity of wash material as the space provided; Group III, double the quantity of wash material, but with V-shaped vents in the putty for escape of excess material. Percentage deviations of the vertical dimensions of stone dies with respect to the master model were significantly different between Groups I and II (the latter being shorter) (p < 0.05). For horizontal dimensions, differences were less consistent, although the deviations for Groups I and II and Groups II and III, with respect to the master model, were significantly different from each other for two of the three dimensions measured (Group II inter-abutment distances were generally larger) (p < 0001 and p < 0.05, respectively). It is concluded that putty recoil, resulting from compression by excess wash material, plays a significant role in the undersizing of working dies, although the level of clinical relevance is less clear. PMID- 14737792 TI - A comparison of three hard chairside denture reline materials. Part I. Clinical evaluation. AB - Three chairside reline materials were each used to reline ten complete dentures. Comfort, fit, colour change, staining and integrity were assessed at 3, 6, and 12 months post-placement. Working and setting times were obtained using an oscillating rheometer. Tokuso Rebase Fast Set had the shortest working and setting times. It was the most colour stable, stain resistant and durable. Coe Kooliner was difficult to use due to high flow. Total Hard had the longest working and setting times, and the poorest stain resistance and durability. Severe loss of reline materials only occurred 12 months after placement, and then mostly from within lower dentures. No differences between the materials were noted for the other parameters investigated. PMID- 14737793 TI - A comparison of three hard chairside denture reline materials. Part II. Changes in colour and hardness following immersion in three commonly used denture cleansers. AB - This study investigated the effects of daily use of three denture cleansers on three hard chairside reline materials over a 6-month period. Two controls were used; one dry and one involving storage in water. Colour and hardness were measured at 0 (baseline) and 6-months. There was no significant colour change with Tokuso Rebase Fast Set. Coe Kooliner and Total Hard changed colour more with the short acting peroxide cleanser (Steradent 3 Minutes) than with the overnight peroxide cleanser (Steradent Fresh) and the hypochlorite cleanser (Dentural). The hardness of all materials remained unchanged over the whole test period. PMID- 14737794 TI - Reducing microleakage in composite resin restorations: an in vitro study. AB - The purpose of this in vitro study was to evaluate the effect of a resin lining on microleakage around composite resin restorations. Standard slot cavities were prepared on the approximal surfaces of 48 extracted human molar teeth. The cavities were restored with two conventional or two condensable composite resins, placed with or without a flowable composite resin lining. The teeth were subjected to thermal stress and immersed in dye solution. Dye penetration was viewed with both a stereomicroscope and a scanning electron microscope. Flowable resin lining material under condensable composites may be beneficial in reducing microleakage around cervical margins. PMID- 14737795 TI - Quality of materials supplied to dental laboratories for the fabrication of cobalt chromium removable partial dentures in Ireland. AB - The adequacy of prescription for fabrication of cobalt chromium removable partial dentures is often regarded as being less than ideal. This study examines the nature and quality of written instructions and master impression sent to dental laboratories in Ireland for fabrication of cobalt chromium removable partial denture frameworks. Questionnaires were issued to dental laboratories seeking specific information relating to the materials (impression materials and trays) and written instructions supplied, as well as the technicians' opinion regarding the suitability of these materials. One hundred completed questionnaires were returned. One-third of master impressions were made using a plastic stock tray and alginate; technicians felt that one-fifth of master impressions were unsuitable; almost three-fifths of written instructions were inadequate. The quality of clinical information examined was found to be less than adequate. PMID- 14737796 TI - Health teams take to the road to update patients with spinal injuries. PMID- 14737797 TI - Contraceptive choices that work. PMID- 14737798 TI - Raising awareness and reducing the risk of needlestick injuries. AB - More than 100,000 needlestick injuries occur in the UK each year. Preventive strategies include training, education, modification of practice, hepatitis B vaccination and the use of needle-protective devices. This paper examines the literature to investigate why injuries occur, why staff do not always report them and what can be done about these problems. PMID- 14737799 TI - Achieving a safer health service: Part 1. Making safety a way of life. AB - Patient safety incidents are a major problem in the health service and it is acknowledged that blaming and reprimanding staff for actual or potential mistakes does not encourage an open and safe culture in which to work. In the first paper in a three-part series the National Patient Safety Agency's seven key to steps to achieving patient safety are discussed. PMID- 14737800 TI - The properties of hydrogel dressings and their impact on wound healing. AB - In recent decades, the management of difficult wounds has been enhanced by the introduction of amorphous hydrogels. This paper examines reports in the literature on the outcomes of studies into the efficacy of this group of dressings, discusses the types of wounds that are most suitable for hydrogel application and poses questions that merit further investigation. PMID- 14737801 TI - A campaign to raise awareness of microalbuminuria screening. AB - Both patients with diabetes and health-care professionals need to be aware that diabetes can cause kidney nephropathy. A simple urine test can detect problems with renal function at a very early stage. The DEMAND campaign has involved diabetes clinic nurses in a study to persuade them to perform this test routinely and to encourage patients to request it. PMID- 14737802 TI - The patient experience of receiving bad news from health professionals. AB - Patients at an NHS trust were questioned about receiving bad news. Despite many positive experiences, problems arose in relation to the setting where the news is delivered and aspects of non-verbal communication, such as the fact that consultants stood when giving the news. This study also examined the patients' interaction with health professionals after the first diagnosis. PMID- 14737803 TI - Reflective thinking: turning a critical incident into a topic for research. AB - Clinical nurses encounter critical incidents every day. While these may be a source of frustration they also have the potential to be turned into research projects so that problems can be examined and others can learn from them. This paper describes the reflective process used to generate a research project from a critical incident encountered in the clinical area. PMID- 14737804 TI - The need to protect children from second-hand tobacco smoke. AB - Second-hand tobacco smoke has serious effects on health--particularly that of children. Voluntary measures to protect people from exposure to second-hand smoke in public places have failed. This paper argues that legislation is needed to ban smoking in public places. PMID- 14737805 TI - A fracture liaison service for patients with osteoporotic fractures. AB - A fracture liaison service, led by a specialist nurse, has been set up in one trust to offer an assessment for osteoporosis to all men and women over 50 with new fractures. The aim is to identify those who have osteoporosis because they are at greatest risk of further fractures. The service delivers strategies to reduce future fracture risk, targeted at those patients at highest risk. PMID- 14737806 TI - How a personal account contributes to nurse knowledge: Rachel's story. AB - This personal account concerns a mother whose sick baby was treated in hospital for a congenital heart anomaly. This mother was inspired by her own experience to train as a children's nurse. Her co-author begins by describing how such personal stories and insights can help raise awareness of parents' feelings and improve nursing practice. PMID- 14737807 TI - Surgical management of aortic and mitral valve disease: an overview. AB - This paper examines aortic and mitral valve disease. It considers how nurses can treat patients with these conditions and provide support and education to help them understand their disease and the risks and benefits of surgery. Nurses also have a role in secondary prevention and in helping patients adapt to living with a prosthetic valve. PMID- 14737808 TI - [Infections with helminths and/or protozoa in cats in animal shelters in the Netherlands]. AB - To determine the prevalence of infections with helminths and protozoa in cats in animal shelters, faecal samples from 305 cats from 22 animal shelters in the Netherlands were examined, using a centrifugation-sedimentation-flotation technique. The association between potential risk factors and the occurrence of an infection was also tested. Infections with helminths and/or protozoa were found in 160 samples (52.5%). Toxocara cati was found in 86 cats (28.2%), Cystoisospora felis in 59 cats (19.3%), Cystoisospora rivolta in 43 cats (14.1%), Capillaria spp. in 34 cats (11.2%), Ancylostoma tubaeforma in 9 cats (3.0%), Taenia taeniaeformis in 9 cats (3.0%), Aelurostrongylus abstrusus in 8 cats (2.6%), Giardia intestinalis in 3 cats (1.0%), Dipylidium caninum in 2 cats (0.7%) and Toxoplasma gondii in 1 cat (0.3%). The highest prevalence was seen in kittens and stray cats. The main preventive factor against infection was a short stay in a shelter. PMID- 14737809 TI - [VWA incidence and crisis organisation]. PMID- 14737810 TI - [More practices on line, better practice]. PMID- 14737811 TI - [WSAVA Congress, Bangkok, October 2003]. PMID- 14737812 TI - [Borderline cases]. PMID- 14737813 TI - [Splenectomy in dogs with immune-mediated hemolytic anemia]. PMID- 14737814 TI - [Misleading clinical experience?]. PMID- 14737815 TI - [A day in the life of a veterinarian]. PMID- 14737816 TI - [Phytotherapy]. PMID- 14737817 TI - [Pseudo-science and illegal remedies]. PMID- 14737819 TI - Staying current in a changing environment. PMID- 14737818 TI - [Homeopathy: evidence-based or not?]. PMID- 14737820 TI - Reconnecting: the client experience of recovery from psychosis. AB - PROBLEM: To understand the experience of recovery from psychosis from the consumer/client perspective. METHODS: A naturalistic, qualitative design with an ethnographic method for data analysis. Subjects (N = 10) were interviewed prior to and during the initial year of treatment with clozapine or risperidone. FINDINGS: Participants described recovery from psychosis as a process that started with improvements in their thinking and feeling, and extended to a series of reconnections with their environment. These reconnections icluded staff and family. Thinking moved from being focused on their internal self to a larger world. CONCLUSION: A person's recovery from pschosis involves the entire self, bringing all components of physical, emotional, mental, and spiritual aspects of themselves into their experiences of life. PMID- 14737821 TI - Gaps in service in the recognition and treatment of depression and suicidal ideation within a four-county area. AB - PURPOSE: To examine gaps in service in the treatment of depression and suicidal ideation. METHODS: A phenomenological investigation of clients' (n = 10) and service providers' (n = 40) perspectives. FINDINGS: Compartmentalized systems of care and lack of continuity between inpatient and outpatient treatment contributed to a sense of frustration in both groups. Receiving mental health care was likened to being given a broken-apart puzzle with individual pieces shuffling around in a box. Little bits of help are available, but none of them mesh. CONCLUSIONS: Comprehensive solutions are needed if the human and social implications of depression are to be addressed. PMID- 14737822 TI - Augmenting treatment of obsessive-compulsive disorder with Antonovsky's sense of coherence theory. AB - TOPIC: The complex nature of obsessive-compulsive disorder (OCD) necessitates a multifaceted approach to treatment. PURPOSE: To examine Antonovsky's sense of coherence theory as a framework for psychotherapy to optimize treatment for individuals with OCD. SOURCES: Selective literature review. CONCLUSIONS: Antonovsky's sense of coherence theory provides a theoretical framework for understanding the multiple pathways by which cognitive behavioral therapy can maintain and improve outcomes for people struggling with OCD. PMID- 14737823 TI - The ABCs of psychopharmacology. AB - The forgoing represents an attempt to reduce psychopharmacology to a presynaptic neuron-synapse-post-synaptic neuron event. Obviously other strategies might be employed resulting in equal or perhaps greater clarity. Limitations of this approach include the lack of discussion about neuronal modulation, second messenger systems (which most NTs discussed here use), expansion of the ion channel discussion, and the implications of excitatory vs. inhibitory physiological effects. Nonetheless, we find this template for understanding psychotropic drug mechanism of action a fruitful lens through which to guide further study of these drugs. We feel like we are moving toward a silver dollar size circle. PMID- 14737824 TI - Treatment considerations for the client with interferon-alpha-induced depression. PMID- 14737825 TI - The rise and fall of the Carbonaria form of the peppered moth. AB - The evidence for change in frequency of the melanic carbonaria morph in the peppered moth Biston betularia (L.) (Lepidoptera: Geometridae) in England and Wales is reviewed. At mid-20th century a steep cline of melanic phenotype frequency running from the north of Wales to the southern coast of England separated a region of 5% or less to west from 90% or more to northeast. By the 1980s the plateau of 90% frequency had contracted to northern England. The frequency has since continued to drop so that the maximum is now less than 50% and in most places below 10%. There have been similar declines in Europe and North America. Evidence from surveys and from two-point records shows the change to require 5% to 20% selection against the melanic. The melanic is more disadvantageous in regions where its frequency was initially high than in regions where it was low. Experiments to investigate predation by birds show a net advantage to carbonaria morphs in regions where typical frequencies were low at the time of the experiment, and a disadvantage where typical frequencies were high. This would be expected if environment and frequency were associated, and selective predation played a part in generating the association. The cryptic advantage of carbonaria was large in areas of heavy pollution where typical frequencies were 20% or less. The moth usually has a low density but is relatively highly mobile. The ability of present information to explain the patterns has been tested in simulations. They indicate a system under strong selection that has always been in a dynamic state without equilibria. PMID- 14737826 TI - Predicting the geography of species' invasions via ecological niche modeling. AB - Species' invasions have long been regarded as enormously complex processes, so complex as to defy predictivity. Phases of this process, however, are emerging as highly predictable: the potential geographic course of an invasion can be anticipated with high precision based on the ecological niche characteristics of a species in its native geographic distributional area. This predictivity depends on the premise that ecological niches constitute long-term stable constraints on the potential geographic distributions of species, for which a sizeable body of evidence is accumulating. Hence, although the entire invasion process is indeed complex, the geographic course that invasions are able to take can be anticipated with considerable confidence. PMID- 14737827 TI - The chickpea, summer cropping, and a new model for pulse domestication in the ancient near east. AB - The widely accepted models describing the emergence of domesticated grain crops from their wild type ancestors are mostly based upon selection (conscious or unconscious) of major features related either to seed dispersal (nonbrittle ear, indehiscent pod) or free germination (nondormant seeds, soft seed coat). Based on the breeding systems (self-pollination) and dominance relations between the allelomorphs of seed dispersal mode and seed dormancy, it was postulated that establishment of the domesticated forms and replacement of the wild ancestral populations occurred in the Near East within a relatively short time. Chickpea (Cicer arietinum L.), however, appears as an exception among all other "founder crops" of Old World agriculture because of its ancient conversion into a summer crop. The chickpea is also exceptional because its major domestication trait appears to be vernalization insensitivity rather than pod indehiscence or free germination. Moreover, the genetic basis of vernalization response in wild chickpea (Cicer reticulatum Ladiz.) is polygenic, suggesting that a long domestication process was imperative due to the elusive phenotype of vernalization nonresponsiveness. There is also a gap in chickpea remains in the archaeological record between the Late Prepottery Neolithic and the Early Bronze Age. Contrary to the common view that Levantine summer cropping was introduced relatively late (Early Bronze Age), we argue for an earlier (Neolithic) Levantine origin of summer cropping because chickpea, when grown as a common winter crop, was vulnerable to the devastating pathogen Didymella rabiei, the causal agent of Ascochyta blight. The ancient (Neolithic) conversion of chickpea into a summer crop required seasonal differentiation of agronomic operation from the early phases of the Neolithic revolution. This topic is difficult to deal with, as direct data on seasonality in prehistoric Old World field crop husbandry are practically nonexistent. Consequently, this issue was hardly dealt with in the literature. Information on the seasonality of ancient (Neolithic, Chalcolithic, and Early Bronze Age, calibrated 11,500 to 4,500 years before present) Near Eastern agriculture may improve our understanding of the proficiency of early farmers. This in turn may provide a better insight into Neolithic agrotechniques and scheduling. It is difficult to fully understand chickpea domestication without a Neolithic seasonal differentiation of agronomic practice because the rapid establishment of the successful Near Eastern crop package which included wheats, barley, pea, lentil, vetches, and flax, would have preempted the later domestication of this rare wild legume. PMID- 14737828 TI - [The mechanisms of development and progression of hypertensive target organ damages]. PMID- 14737829 TI - [Current state and perspective of studies on hypertensive complications]. AB - Studies on cardiovascular complications in hypertensive patients have been extensively progressed in basic and clinical researches. The molecular mechanism of sensing system of mechanical stress have not been fully understood. Rho/Rho kinase and oxidative stress are molecules activated by stretch and are highly possible to contribute to cardiovascular complications. One of clinical progress in this area is the development and wide distribution of noninvasive diagnostic devices for atherosclerosis. Some of these devices are used as an surrogate markers in large scale randomized clinical trials. Currently, new therapeutic targets and strategies have been investigated and tailored medicine using pharmacogenetics is expected in the near future to manage hypertensive complications. PMID- 14737830 TI - [Treatment strategy on hypertensive cardiovascular damage observed in various guidelines]. AB - Since the publication of the Japanese guidelines for the treatment of hypertension in 2000, a lot of large-scale clinical trials on the treatment of hypertension have been reported. Furthermore new guidelines for the treatment of hypertension, such as JNC-VII and ESH-ESC guidelines have been reported in 2003. On the consideration of results of these trials and new guidelines, treatment strategy on hypertensive cardiovascular damage was described in this manuscript. The initiation criteria of drug therapy, modifications of life-style, selection of antihypertensive drugs and target blood pressure are various in each guideline. However, strict control of blood pressure is the most important to protect cardiovascular damages, regardless differences in the treatment strategy in each guideline. In the drug therapy Ca channel blockers, inhibitors of the renin-angiotensin system and diuretics are mainly used as monotherapy or a combination therapy to obtain the target blood pressure. PMID- 14737831 TI - [Angiotensin II in organ organopathy]. AB - Angiotensin II induces the organ derangements is not generated by the so-called classic rennin-angiotensin system but by the tissue angiotensin II generating system. We have confirmed this evidences by using different types of animal models such as hypertension, arteriosclerosis, vascular narrowing by balloon injury and in vein graft disease. In addition, we found that the ACE and chymase activities are increased in human aneurismal aorta. ACE inhibitor is effective to protect the development of hypertension and arteriosclerosis, but not to other models because the angiotensin II produced by chymase is involved in such diseases. Angiotensin II produced separately by ACE and chymase, participates independently in the development of vascular derangements. PMID- 14737832 TI - [Angiotensin II receptor]. AB - The peptide angiotensin(Ang) II exerts hemodynamic, and renal as well as cardiovascular structural effects. Multiple lines of evidence have suggested the existence of several Ang II receptor subtypes. Recent evidence has revealed that the functions of the AT1 and AT2 receptors are mutually antagonistic in various cells and tissues. The effect of AT1 blocker(ARB) may not be entirely due to the blockade of the AT1 receptor. When AT1 receptor is blocked and unbound Ang II may act on AT2 receptor and Ang 1-7 and Ang IV via AT4 receptor might be involved in the effects of ARB. If the AT2 receptor contributes to the pathogenesis and consequent remodeling of cardiovascular diseases in human, ARB may have some specific effects in the treatment of cardiovascular diseases. A more extensive knowledge of the AT2 receptor could therefore contribute to the understanding of the clinical beneficial effects of ARB. PMID- 14737833 TI - [Aldosterone]. AB - Nonepithelial effect of aldosterone has been recognized as a cardiovascular risk factor independent of elevation of blood pressure. Pro-inflammatory effect of aldosterone in vasculature appears to be an initiating event for the target organ injuries. Recent clinical trials of aldosterone receptor blockers in congestive heart failure indicate the efficacy of aldosterone antagonism to prevent cardiac death. In addition to elevated plasma levels of aldosterone, mineralocorticoid receptor expression in cardiovascular tissues may contribute to the progression of heart failure. In this review, emerging role of aldosterone as a cardiovascular risk is highlighted. PMID- 14737834 TI - [Rho-Rho kinase pathway]. AB - The serine/threonine protein kinase Rho kinase, a downstream effector of the small GTPase Rho, regulates actin cytoskeletal organization, cell adhesion, cell motility, smooth muscle contraction and gene expression. Rho kinase constitutes the essential regulatory system for myosin phosphatase, controlling myosin light chain phosphorylation and thereby vascular tone. Inhibitors of Rho kinase were shown to be effective in ameliorating hypertension, vasospasm, vascular injury and associated organ damages in various animal disease models. These recent observations imply potential usefulness of the Rho kinase inhibitors as novel therapeutic and preventive agents for vascular disorders and accompanying organ injury. PMID- 14737835 TI - [Oxidative stress, reactive oxygen species]. AB - Hypertension is a risk factor for cardiovascular and cerebrovascular outcome. Hypertension is associated with oxidative stress. Alteration in endothelial function is an initial step in the pathogenesis of atherosclerosis. A balance between ambient levels of super oxide and released nitric oxide(NO) plays an important role in the maintenance of endothelial function. It is well known that reactive oxygen species, including hydroxy radicals, directly scavenge NO and produce toxic peroxynitrite. Angiotensin II and mechanical stress generate the reactive oxygen species through the activation of NADH/NADPH oxidase in hypertension. Several investigators have shown that oxidative stress is involved in enhanced vascular growth, vascular inflammation, and impaired endothelium dependent in hypertension. In this review, we would like to explain the role of oxidative stress in hypertensive organ damages. PMID- 14737836 TI - [Significance of insulin resistance to atherosclerotic complications in essential hypertension]. AB - Insulin resistance and compensatory hyperinsulinemia are recognized not only in type 2 diabetes mellitus(DM) but also in essential hypertension(EHT), hyperlipidemia and obesity; these are known as the components of metabolic syndrome and accumulation of these components increase risk of cardiovascular diseases(CVD). When coronary angiographic findings were evaluated in patients with coronary artery disease(CAD), the severity was higher in CAD with DM than that without DM. Even in CAD without DM, the severity of coronary angiographic findings was higher in CAD with insulin resistance than that without insulin resistance. When residents of rural communities in Japan were followed 8 years, the incidence of CVD was 3.5 times higher in subjects with insulin resistance than those without insulin resistance. One of the intracellular signal transduction of insulin receptor; MAP kinase may be concerned atherosclerotic mechanisms of insulin resistance. These findings suggest that insulin resistance is a significant background of atherosclerosis, and insulin resistance is one of the major facilitation factors of genesis and progression of CVD. PMID- 14737837 TI - [klotho gene]. AB - The klotho gene, originally identified by insertional mutagenesis in mice, suppresses multiple aging phenotypes(e.g. arteriosclerosis, pulmonary emphysema, osteoporosis, infertility, short lifespan). We have shown that mice deficient for the klotho gene show endothelial dysfunction as manifested by an attenuated response of aortic relaxation in response to acetylcholine stimulation. Nitric oxide production was also significantly reduced in klotho deficient mice. A decrease in klotho gene expression in animals under sustained circulatory and metabolic stress(e.g. atherosclerosis). The klotho gene delivery improves endothelial dysfunction through a pathway involving nitric oxide, and is involved in modulating vascular function(e.g. hypertension, vascular remodeling). Our findings establish the basis for the therapeutic potential of klotho gene delivery in atherosclerotic disease. PMID- 14737838 TI - [Diagnosis of left ventricular diastolic dysfunction by Doppler echocardiography]. AB - Left ventricular(LV) diastolic dysfunction with preserved LV systolic function is common among patients with hypertension, especially with LV hypertrophy. Doppler echocardiography is one of the most useful clinical tools for the evaluation of diastolic function. Mitral inflow and pulmonary venous flow velocities are used not only for the assessment of diastolic function but also for predicting prognosis. Recently, tissue Doppler echocardiography has been also applied to evaluate diastolic function. Accurate assessment of diastolic function has been demonstrated by measuring both mitral annulus and mitral inflow velocity. In this article, We review the diagnosis of diastolic dysfunction by Doppler echocardiography using mitral inflow velocity, pulmonary venous flow velocity and mitral annulus velocity measured by tissue Doppler imaging. PMID- 14737839 TI - [Arteriosclerosis and pulse wave velocity]. AB - Impairment of the arterial compliance or loss of Windkessel effect of elastic arteries causes increased afterload to the heart and increased pulsatile flow to the peripheral vasculatures. The former induces left ventricular hypertrophy or dysfunction and the latter induces small vessel damage or end organ dysfunction. Thus, the arterial compliance plays important roles in the course of hypertension. Therefore; it is worthwhile to measure the elastic properties of aortoarterial system in patient with hypertension. The velocity of the pressure wave along an arterial system, known as pulse wave velocity(PWV), is related to the average stiffness of an arterial segment between measurement sites. The measurement of PWV is inversely related to arterial wall distensibility, which offers a simple and potential approach. There are numerous reports which PWV is a forceful marker and predictor of the cardiovascular risk in hypertensive or other arteriosclerotic disorders. Thus, PWV measurement is recommended in patients with hypertension for early detection of organ damages or estimation of the cardiovascular risk, as well as for the evaluation of the effectiveness of the treatment as a surrogate marker. PMID- 14737840 TI - [Evaluation of carotid artery lesion by echography]. AB - The recent advance of the carotid artery echotomography indicates the intima media thickness(IMT) of the carotid artery as one of surrogate endpoints of atherosclerosis in subjects with multiple risk factors, such as hypertension, hyperlipidemia, and diabetes mellitus. IMT is shown to be increased in subjects with hyperlipidemia, type 1 and 2 diabetes mellitus, and other several diseases. Also, increase in IMT is related with the prevalence of cerebral infarction and coronary artery diseases. The carotid artery plaques including soft or calcified plaques were shown to predict appearance of strokes. Several drugs, such as anti diabetogenic drugs(insulin-sensitizer, biguanides, and alfa-glucosidase inhibitor), hypotensive drugs(ACEI, Ca-blocker), and anti-platelet drugs were shown to attenuate the progression of IMT. Recently, we have shown that an anti platelet drug arrested the progression of IMT and significantly reduced appearance of asymptomatic cerebral infarction in subjects with type 2 diabetes. These data clearly indicate the usefulness of the carotid artery echography in subjects with atherosclerosis and IMT could evaluate the effects of treatment for atherosclerosis. PMID- 14737841 TI - [Microalbuminuria in hypertension]. AB - Microalbuminuria is a sensitive marker of early renal damage in essential hypertension as well as diabetes mellitus. Elevated urinary albumin excretion has positive correlations with other cardiovascular risk factors. Microalbuminuria has an independent value to predict cardiovascular disease in the patients with not only diabetes mellitus but also with essential hypertension and even in general population. The seventh report of the joint national committee on prevention, detection, evaluation and treatment of high blood pressure (JNC 7) has adopted microalbuminuria or estimated glomerular filtration rate < 60 ml/min as one of the major cardiovascular risk factors. It is an important future task whether decrease in urinary albumin excretion with treatment improves prognosis of individual patients. PMID- 14737842 TI - [NOS gene polymorphism]. AB - Nitric oxide(NO), a potent vasodilator produced by endothelial cells, plays an important role in the regulation of blood pressure and regional blood flow, and also inhibits platelet aggregation and leukocyte adhesion to vascular endothelium and inhibits vascular smooth muscle cell proliferation. Various genetic polymorphisms of the endothelial nitric oxide synthase(eNOS) gene have been reported as susceptibility genes in a number of cardiovascular diseases. The functional significance of these polymorphisms has not yet been demonstrated, but recent study revealed that eNOS Glu298Asp variant is likely to have a functional effect on the eNOS protein. The missense Glu298Asp variant was correlated with increased coronary spasm, myocardial infarction, essential hypertension, left ventricular hypertrophy, carotid atherosclerosis, and cerebral infarction. PMID- 14737843 TI - [Genes in rennin-angiotensin system]. AB - A large number of association studies revealed the importance of gene polymorphisms in renin-angiotensin system(RAS) in the pathogenesis of hypertension. Among these many polymorphisms of RAS genes, M235T or G-6A polymorphism of angiotensinogen gene increases the risk for salt sensitivity, lacunar infarction, non-dipper type of blood pressure variation, etc. and plasma angiotensinogen level. On the other hand, homozygous deletion polymorphism of angiotensin converting enzyme gene(ACE DD) increases the risk for male hypertension, ischemic heart disease and renal disease via increase of plasma and tissue ACE levels, and shows poor response to medication. Gene targeting experiments in mice enable us to examine whether quantitative changes in gene expression in RAS affect blood pressure, resulted that a simple computer simulation concerning RAS and the kallikrein-kinin system(KKS) emphasize the importance of interaction between RAS and KKS. This mini-review also describes the effect of renin or AT1 receptor gene polymorphisms, and the role of RAS genes in the correlation between inflammation and atherosclerosis. PMID- 14737844 TI - [Prevention and treatment of the hypersensitive organopathy--the necessity of cardiovascular protection]. AB - In our country, cardiovascular disease, such as ischemic heart disease and apoplexy, occupies the epistasis of mortality rate. One of the causes of cardiovascular disease is arteriosclerotic change and hypertension. It is a common understanding that the prevention of the arteriosclerotic change and antihypertensive treatment are very important. Lately, it is found angiotensin converting-enzyme inhibitor has anti-arteriosclerotic action that cannot be explained only in a blood pressure control. Reexamination of the strategy is needed in antihypertensive treatment. The close relation exists between antihypertensive treatment and cardiovascular protection. The cardiovascular protection is important in the new antihypertensive therapy. PMID- 14737845 TI - [Anti-hypertensive drugs associated with the improvement of endothelial function]. AB - Vascular endothelium is not merely a barrier between circulating blood and tissue, but also regulates vascular tone through the secretion of numerous vasoactive substances in response to hemodynamic changes. Thus endothelial injuries by hypertension and atherogenic risk factors play important pathogenic roles in the vascular disease state, and the protection of endothelial function leads to amelioration of progressive vascular disease. In this review, we describe the pathophysiology of endothelial dysfunction in vascular disease, and therapeutic significance of vaso-protective agents, especially in the cotex of endothelial protection. PMID- 14737846 TI - [Therapeutic strategy for the treatment with hypertensive renal injury]. AB - Hypertension constitutes a pivotal determinant of the progression of renal disease, which would raise the risk for cardiovascular events. From the standpoint of renal micro-circulation, correction of glomerular hypertension could retard the development of renal injury, which is attainable by the reduction in renal efferent arteriolar resistance as well as systemic blood pressure. Although both ACE inhibitors and angiotensin receptor blockers are established as a tool for improving glomerular hypertension, whether calcium antagonists ameliorate this abnormality remains unclear. However, recent clinical trials including ALLHAT and INSIGHT demonstrate a beneficial action of amlodipine and nifedipine on the development of renal injury. Therefore, calcium antagonists can be used not only as a first line drug, but also as add-on therapy that could potentiate the hypotensive action of underlying medication. PMID- 14737847 TI - [Anti-hypertensive treatment for primary and secondary prevention of brain attack and failure]. AB - Although anti-hypertensive treatment is known to be effective for preventing first-ever stroke, there has been a long debate, whether this treatment is safe and effective for preventing recurrent stroke and subsequent disabled conditions. PROGRESS trial was designed to clarify these concerns about the efficacy and safety of blood-pressure-lowering treatments for the chronic stroke patients and clearly showed that anti-hypertensive treatment should be quite safe and effective for preventing recurrent stroke. The results of other clinical trials such as HOPE, Syst-Eur2 and SCOPE also suggested the at most importance of anti hypertensive treatment for primary prevention of cognitive impairment and dementia. In this review, the significance of recent clinical trials were fully discussed in relation to the hypertension-induced pathophysiological changes of the brain. PMID- 14737848 TI - [Treatment for hypertensive patients with diabetes mellitus]. AB - The frequency of diabetes and hypertension is increasing worldwide. Diabetes mellitus doubles the risk of cardiovascular diseases, even in hypertensive patients who are already at high risk because of their high blood pressure. Combination of 2 or more drugs is usually needed to achieve the target BP goal of less than 130/85 mmHg. Thiazide diuretic, beta-blockers, ACE inhibitor, ARBs and Ca blockers are beneficial in reducing cardiovascular events. However, the ACE inhibitors- or ARB-based treatments favorably affect the progression of diabetic nephrology and reduce albuminuria. PMID- 14737849 TI - [Cardiovascular protective action of Rho-kinase inhibitors]. AB - The Rho/Rho-kinase pathway plays an important role in pathophysiology and formation of hypertensive vascular diseases. This pathway contributes to both increased vascular resistance and atherosclerotic changes. The latter includes inflammatory and proliferative changes. In patients with hypertension, inhibition of Rho-kinase has been shown to decrease the increased vascular resistance. Therefore, Rho-kinase will be a new therapeutic target for hypertension and hypertensive vascular diseases. Thus, Rho-kinase inhibitors may have a cardioprotective action. PMID- 14737850 TI - [Anti-inflammatory effect of HMG-CoA reductase inhibitors]. AB - It has been suggested that HMG-CoA reductase inhibitors(statins) have additional cholesterol-independent pleiotropic effects on preventing coronary events and strokes. One of major pleiotropic actions of statins, which have been proposed, is anti-inflammatory effect. Statins have been shown to reduce infiltration of inflammatory cells into atherosclerotic lesions. It has been also reported that statins increase production of nitric oxide, reduce expression of proinflammatory cytokines and adhesion molecules, and lower plasma C-reactive protein levels. Although these effects may partially account for anti-inflammatory property of statins, its mechanisms are not fully understood. Therefore, further studies are expected to elucidate process of anti-inflammatory effect of statins. PMID- 14737851 TI - [Anti-atherosclerotic effects of PPAR gamma ligands]. AB - Peroxisome proliferator-activated receptor gamma(PPAR gamma) belongs to a nuclear receptor super family which is activated by insulin sensitizer, thiazolidinediones. The expression of PPAR gamma was detected in the vascular tissues and PPAR gamma ligands have various effects on all cells which constitute the vasculature including endothelial cells, vascular smooth muscle cells and monocytes/macrophages. The net effect by PPAR gamma ligands is proven to be anti atherosclerosis. In various atherosclerogenic mouse models and balloon injury model, PPAR gamma ligands attenuate atherosclerotic lesion formation and intimal hyperplasia. In human study, various beneficial effects by PPAR gamma ligands were reported in terms of atherogenesis. PPAR gamma ligands attenuate the increase in intima-media thickness in diabetic patients. However, long-term effects remain to be seen. PMID- 14737852 TI - [Lipoxygenase inhibition and protection of cardiovascular system]. AB - The arachidonic cascade involves three types of metabolic pathways; cyclo oxygenase, lipoxygenase(LO), and cytochrome P450. The products of LO pathway participate in the pathogenesis of variety of disease such as allergic diseases and hypertension. In particular, 12-hydroxyeicosatetraenoic acid(12-HETE); product of 12-LO pathway, is concerned in the development of hypertension induced by angiotensin-II. In fact, several investigators have reported that 12-HETE has a critical role in hypertension and LO inhibitors have antihypertensive effects in experimental animals. And so, we can expect organ-protective effects of LO inhibitors as well as antihypertensive effects. Besides 12-LO, several investigators mentioned that 5- and 15-LO also have influences to cardiovascular systems. Therefore, we can expect further elucidation of the mechanism of 12-LO's participation in the organ damage and the clinical roles of 12-LO inhibitors to prevent from organ failure in future. PMID- 14737853 TI - [Clinical profiles of new angiotensin II receptor blockers: telmisartan, olmesartan medoxomil, and irbesartan]. AB - Clinical use of type 1 angiotensin II receptor blockers(ABRs) is rapidly increasing because of their high safety as well as excellent efficacy. Recent clinical trials have demonstrated that telmisartan at a daily dose of 20-80 mg, olmesartan medoxomil at 10-40 mg, and irbesartan at 150-300 mg are effective and safe for the treatment of essential hypertension, severe hypertension and hypertension associated with renal diseases. These ARBs are similar to ACE inhibitors in terms of antihypertensive efficacy, but lack the adverse effect of cough. Long-term effects should be compared among ARBs, ACE inhibitors, and other antihypertensive drugs. PMID- 14737854 TI - [Effect of angiotensin II receptor antagonist on heart failure]. AB - An angiotensin II receptor antagonist, candesartan has been shown to improve left ventricular dysfunction and exercise tolerance. The assessment of response to candesartan heart failure in Japan(ARCH-J) is a randomized, double-blind, placebo controlled 6-month study enrolled 305 patients who has not been previously treated with ACE inhibitors, who had not been adequately controlled with ACE inhibitors or who were intolerant of ACE inhibitors. The incidence of confirmed progression of heart failure was significantly lower in the candesartan group(7.4%) than in the placebo group(22.2%), with risk reduction 63.8. Cardiovascular events were also significantly lower during treatment with candesartan than with placebo(10.8% vs 22.9%) with risk reduction of 50.2%. ARCH J study showed that candesartan cilexetil, 8 mg/day, significantly improved the progression of heart failure when compared with placebo. PMID- 14737855 TI - [Benefits of candesartan in the treatment of symptomatic heart failure--CHARM programme]. AB - The CHARM programme, which recruited 7,601 patients, compared the angiotensin receptor blocker, candesartan, with a placebo in three different populations with class II-VI heart failure. Candesartan reduced cardiovascular mortality and/or hospitalization for heart failure in patients with heart failure and ejection fractions of greater than 40% or less, and total mortality was reduced in these patients as well. Similar benefits were seen whether or not background therapy with ACE inhibitors, beta blockers', or spironolactone was used. Pooled analysis of the three studies showed that candesartan provided a significant reduction in cardiovascular death and also demonstrated a positive trend in the overall reduction in all cause mortality. The major adverse effects were hyperkalemia, increase in creatinine concentration and hypotension. PMID- 14737856 TI - [Treatment of heart failure in the 21st century--the position of ARB in view of therapy and preventive medicine: discussion]. PMID- 14737857 TI - [Child neurology in borderless age]. PMID- 14737858 TI - [A new challenge for activities in the Japanese Society of Child Neurology. Interviw by Ikuro Konishi]. PMID- 14737859 TI - [Epidemiology of subacute sclerosing panencephalitis in Okinawa, Japan--the second report 1977-1999]. AB - There were 16 cases (11 males and 5 females) of SSPE in Okinawa from 1977 to 1999. The incidence was 0.58 per million population per year for the last 23 years, being higher than in other reports in Japan. Six of the 16 cases contracted measles in 1990. The measles antibody of SSPE cases after 1989 became lower than previously. PMID- 14737860 TI - [Adaptive behavior scale for persons with severe physical and mental disability]. AB - An adaptive behavior scale for persons with severe physical and mental disability was proposed. This scale consisted of 43 items from five areas; interpersonal relationship, perception, expression, interest and play, and daily life. Each item was scored on a 0-2 scale. Eighty-two persons (52 males, 30 females) with severe physical and mental disability, aged 1-49 (mean 20.2) years, were examined with this scale. They were institutionalized or attended a day care center. The rating for each item was 1.60-0.09 (mean 0.75). The Cronbach's alpha value was calculated to be 0.95, indicating good internal consistency. For 39 persons attending a center, estimates were consistent between the mother and care staff on only 61% of the items, showing insufficient inter-rater reliability. On the whole, this scale is useful in assessing adaptive behaviors of the subjects. PMID- 14737861 TI - [Memory functions in children with attention deficit/hyperactivity disorder--the effects of methylphenidate on them]. AB - The memory functions or capacities in attention-deficit/hyperactivity disorder (AD/HD) are still not clear, though it has been pointed that the working memory in AD/HD could be impaired due to difficulties of motor inhibition or self regulation. We examined the Auditory Verbal Learning Test (AVLT) and the Rey Osterrieth Complex Figure Test (RCFT) in addition to the memory tasks of ordinary intelligence tests (WISC-III and K-ABC) in children with AD/HD. Whether these results could be improved by methylphenidate administration or not was also evaluated. Over the half cases had normal results without medication. Some cases in whom methylphenidate were clinically effective showed improved memory functions, especially in the auditory long-term memory, after methylphenidate administration. In conclusion, memory capacities seem normal in AD/HD. Methylphenidate does not have an effect on the memory capacities, but may improve the strategies in which the short-term memory can be effectively transferred to the long-term memory. PMID- 14737862 TI - [Surgical procedure for gastroesophageal reflux disease in patients with severe motor and intellectual disabilities: problems and prognosis]. AB - Gastroesophageal reflux disease (GERD) is a complications of patients with severe motor and intellectual disabilities. We examined on 17 patients who operated with GERD. They were divided into 2 groups. Group 1 was comprised of young cases with severe spasticity and chronic respiratory insufficiency. They were under 23 years old. Group 2 with severe intellectual disabilities, aerophagia and/or rumination. They were around 30 years old. Older patients had deformities of the stomach and esophagus. After the operations, fourteen patients had a fair prognosis, three had persistent gastroesophageal reflux (GER), and six patients had subsequent relapse of their GER, and two died. PMID- 14737863 TI - [Serial median nerve SEPs and SSEPs in patients with West syndrome]. AB - We studied serial median nerve somatosensory evoked potentials (SEPs) and short latency somatosensory evoked potentials (SSEPs) in 17 patients with West syndrome. Four of the 7 patients with absent SEPs in the initial examination showed recognizable SEPs in the follow-up studies, associated with improvement of electroencephalogram (EEG). This indicated that SEPs were variable with condition of epilepsy and lack of initial SEPs was not always a poor prognostic factor for seizure control and developmental outcome. Persistent lack of SEPs, however, indicated poor outcome of seizures, EEG and development. Central conduction time in SSEPs did not correlate with seizure or developmental outcome. PMID- 14737864 TI - [Evaluation of sympathetic skin response in patients with attention deficit/hyperactivity disorder presenting with comorbid disorders]. AB - We studied sympathetic skin response (SSR) to visual emotional stimuli in two children with attention deficit/hyperactivity disorder (AD/HD) and nine healthy controls, and correlated them with comorbid disorders. They were diagnosed as having conduct disorder at administration. Two years after interventions of medical care, counseling and education, they were improving in behavior, one with oppositional defiant disorder (ODD) and another without ODD at this study. SSR were evoked in the patient without ODD (a 10-year-old boy), but not in the patient with ODD (a 12-year-old boy). These findings indicate failure of a patient with both AD/HD and ODD to respond autonomically to social stimuli, and suggest association between emotional signal and empathy. PMID- 14737865 TI - [Immunohistochemical studies of a variant of congenital muscular dystrophy]. AB - Three Japanese patients from 2 families had a phenotype indistinguishable from that of Fukuyama-type congenital muscular dystrophy (FCMD). A full mutational analysis of the fukutin gene, however, revealed neither a 3 kb insertion (the Japanese founder mutation) nor a point mutation. A RT-PCR analysis of one of the patients revealed a normal expression of the fukutin transcript, suggesting that they have a new variant of CMD. An immunohistochemical analysis of the muscle of one case showed that the immunoreaction to alpha-dystroglycan (DG) was barely detectable on the surface membranes of muscle fibers. Immunoreactions to beta-DG, dystrophin, laminin alpha-2 chain and sarcoglycan were normal. These findings raise the possibility that the abnormality of alpha-DG is integral to the pathology seen in this variant of CMD. Analysis of POMGnT1 gene, which is causative of muscle-eye-brain disease, revealed no mutation in this case. PMID- 14737866 TI - [A case of chronic cerebellitis with anti-glutamate receptor delta 2 antibody]. AB - A 1-year-8-month-old patient developed cerebellar ataxia following a prodromal infection. Despite initial diagnosis of acute cerebellar ataxia, his symptoms lasted for more than 30 days. High-dose intravenous immunoglobulin and steroid pulse therapy failed to ameliorate his cerebellar symptoms, which fluctuated in association with infections. At the age of 3 years and 8 months, he had mental retardation with cerebellar symptoms. Findings of MR imaging and single photon emission computed tomography were normal. Neuron-specific enolase of cerebrospinal fluid (CSF) ranged from 10.4-17.6 ng/ml, correlating with the cerebellar symptoms. Serum and CSF anti-glutamate receptor delta 2 antibodies were detected in the serum and CSF. We diagnosed him as having chronic cerebellitis associated with anti-glutamate receptor delta 2 antibody. PMID- 14737867 TI - [A case of bilateral paramedian thalamic infarction in childhood with the sensory disturbance and the sensory loss of taste]. AB - Bilateral paramedian thalamic infarcts are characterized by disturbance of consciousness, followed by persisting dementia, decreased spontaneity, apathy, amnesia and paralysis of eye movement. We report a 15-year-old boy with this syndrome, who exhibited transient coma at the onset. In addition to the typical symptoms, he complained of sensory disturbance in the lower extremities and face and the loss of taste sense. MRI showed symmetric paramedian thalamic infarction. There was no lesion in the midbrain. The etiology of infarct in this boy remained unknown despite extensive laboratory and neuroradiological examination. His sensory disturbance in the extremities and face may be due to extensive involvement of the inferolateral area of the thalamus by infarction of the paramedian thalamic artery. This patient illustrates that bilateral paramedian thalamic infarction can occur in a previously healthy child. PMID- 14737868 TI - [Five patients with subacute sclerosing panencephalitis treated with intraventricular alpha-interferon and inosinpranobex]. AB - We followed up 5 patients with subacute sclerosing panencephalitis (SSPE) for 14 to 81 months. They were treated with alpha-interferon (INF-alpha) and oral inosinpranobex (INP) in an early stage of Jabbour stage II and within 5 months after the onset. On admission, Ommaya reservoir was implanted for the intrathecal administration of INF-alpha. The dose was 1 x 10(5) U/m2 initially and daily increased to 1 x 10(6) U/m2. A total dose of 30 x 10(6) U/m3 was given to them over a 4-weeks to 6-weeks period. After discharge, a dose of 15 x 10(6) U/m2 in three patients was given weekly and a dose of 30 x 10(6) U/m2 in the other patients. In addition, all patients received oral INP. One patient showed mild progression and remained in early stage of Jabbour stage II. In the remaining 4 patients, the disease progressed to Jabbour stage III. Despite the small number of patients studied here, the results suggest that treatment with INF-alpha plus oral INP is ineffective in an early stage of SSPE. PMID- 14737869 TI - [Improvement of action myoclonus in a patient with dentatorubral-pallidoluysian atrophy by piracetam]. AB - We report a 13-year-old girl with dentatorubal-pallidoluysian atrophy (DRPLA), presenting clinically as progressive myoclonic epilepsy. The action myoclonus, which severely impaired her daily life, was markedly improved by administration of piracetam, a drug reportedly useful for myoclonus of cortical origin. In our case, piracetam effectively suppressed severe subcortical myoclonus of DRPLA, suggesting that the drug may be useful in the treatment of both cortical, and subcortical myoclonus. PMID- 14737870 TI - [Relationship between acute encephalopathy and febrile convulsions]. PMID- 14737871 TI - Explanations change, therapies remain the same for the time being.... PMID- 14737872 TI - Re: Yatani et al. Comparison of sleep quality and clinical and psychologic characteristics in patients with temporomandibular disorders 2002;16:221-228. PMID- 14737873 TI - Interventions for the treatment of burning mouth syndrome: a systematic review. AB - AIMS: To carry out a systematic review of previous studies to determine the effectiveness of any intervention vs placebo for relief of symptoms and improvement in quality of life of patients with burning mouth syndrome (BMS) and to assess the quality of the studies. METHODS: Electronic databases, conference proceedings, and bibliographies of identified publications were searched (up to September 2001) to identify relevant literature, irrespective of language of publication. Randomized controlled trials and controlled clinical trials of interventions used for the treatment of BMS in comparison to a placebo were included. The primary outcome was relief of burning/discomfort. The screening of studies, validity assessment, and data extraction were undertaken independently and in duplicate. Since statistical pooling of data was inappropriate, a qualitative assessment was undertaken. RESULTS: Seven trials, evaluating antidepressants, cognitive behavioral therapy, analgesics, hormone replacement therapy, and vitamin complexes, met the inclusion criteria. None of the trials was able to provide conclusive evidence of effectiveness. However, cognitive behavioral therapy may be beneficial in reducing the intensity of the symptoms. CONCLUSION: Given that the research evidence is, as yet, unable to provide clear, conclusive evidence of an effective intervention, clinicians need to provide support and understanding when dealing with BMS sufferers. Psychological interventions that help patients to cope with symptoms may be of some use, but promising and new approaches to treatment still need to be evaluated in good quality randomized controlled trials. PMID- 14737874 TI - Pharmacologic interventions in the treatment of temporomandibular disorders, atypical facial pain, and burning mouth syndrome. A qualitative systematic review. AB - AIMS: To carry out a systematic review of the literature in order to assess the pain-relieving effect and safety of pharmacologic interventions in the treatment of chronic temporomandibular disorders (TMD), including rheumatoid arthritis (RA), as well as atypical facial pain (AFP), and burning mouth syndrome (BMS). METHODS: Study selection was based on randomized clinical trials (RCTs). Inclusion criteria included studies on adult patients (> or = 18 years) with TMD, RA of the temporomandibular joint (TMJ), AFP, or BMS and a pain duration of > 3 months. Data sources included Medline, Cochrane Library, Embase, and Psych Litt. RESULTS: Eleven studies with a total of 368 patients met the inclusion criteria. Four trials were on TMD patients, 2 on AFP, 1 on BMS, 1 on RA of the TMJ, and 3 on mixed groups of patients with TMD and AFP. Of the latter, amitriptyline was effective in 1 study and benzodiazepine in 2 studies; the effect in 1 of the benzodiazepine studies was improved when ibuprofen was also given. One study showed that intra-articular injection with glucocorticoid relieved the pain of RA of the TMJ. In 1 study, a combination of paracetamol, codeine, and doxylamine was effective in reducing TMD pain. No effective pharmacologic treatment was found for BMS. Only minor adverse effects were reported in the studies. CONCLUSION: The common use of analgesics in TMD, AFP, and BMS is not supported by scientific evidence. More large RCTs are needed to determine which pharmacologic interventions are effective in TMD, AFP, and BMS. PMID- 14737875 TI - Symptoms, signs, and clinical diagnoses according to the research diagnostic criteria for temporomandibular disorders among Finnish multiprofessional media personnel. AB - AIMS: To apply the Finnish version of the Research Diagnostic Criteria for Temporomandibular Disorders (RDC/TMD) Axis I to assess the occurrence of symptoms, signs, and specific subgroups of TMD, and to study the associations between the most common diagnoses and categoric demographic characteristics (gender, age group, marital status, type of work). METHODS: All 30- to 55-year old employees of the Finnish Broadcasting Company with at least 5 years at their current employment received postal questionnaires (n = 1784). Of the 1339 respondents (75%), a randomly selected one fifth were clinically examined according to the RDC/TMD Axis I (n = 241, males 48%). RESULTS: Pain symptoms in the face or jaw regions were perceived by 14.9% and pain with 1 or more jaw movements by 9.1%. Diagnoses by the RDC/TMD criteria were: Group I: myofascial pain in 12.9%, myofascial pain with limited opening in 0.4%; Group II: disc displacement with reduction in the right temporomandibular joint (TMJ) in 9.1% and in the left TMJ in 10.8%; Group III: arthralgia in 0.4% and 0.8%, osteoarthritis in 0% and 0.4%, and osteoarthrosis in 1.2% and 1.2%, respectively, in the right and left TMJs. The most common diagnoses were found more often among women than among men. No TMD diagnosis based on the RDC/TMD was obtained in 73% of the subjects. CONCLUSION: The RDC/TMD appear to be of benefit in diagnosing TMD among these multiprofessional media personnel and thus may be suggested for use among nonpatient populations. PMID- 14737876 TI - The effect of glutamate-evoked masseter muscle pain on the human jaw-stretch reflex differs in men and women. AB - AIMS: To compare jaw-stretch reflex responses in male and female subjects and to determine whether injection of glutamate into the masseter muscle facilitates these responses in both sexes. METHODS: Jaw-stretch reflex responses were evoked with a muscle stretcher, and pain intensity was scored by 11 men and 13 women before and after the injection of glutamate (1.0 mol/L, 0.2 ml) into the masseter muscle. The subjects rated glutamate-evoked pain intensity on a visual analog scale. RESULTS: Baseline jaw-stretch reflex responses were larger and glutamate injections into the masseter muscle were significantly more painful in women than in men, however, glutamate significantly facilitated jaw-stretch reflex responses in men but not in women. CONCLUSION: These results suggest that there is a significant sex-related difference in human jaw-stretch reflex responses and their modulation by painful stimuli. Since one possible function of facilitated jaw-stretch reflex responses during jaw muscle pain may be to reduce jaw mobility and thus protect against further exacerbation of an existing injury, the finding of a sex-related difference in modulation of jaw-stretch reflex responses may prove to be important in clarifying why the prevalence of temporomandibular disorders is greater in women than in men. PMID- 14737877 TI - Pressure pain thresholds in the craniofacial region of female patients with rheumatoid arthritis. AB - AIMS: To determine the temporomandibular joint (TMJ) pressure pain threshold (PPT) in female patients with rheumatoid arthritis (RA) and TMJ involvement in comparison with healthy females, in order to determine its clinical usefulness for local pain assessment. METHODS: Forty-two female patients with the diagnosis of RA, 17 of them positive and 25 negative for rheumatoid factor were investigated, as well as 17 healthy females. A pressure algometer was used to assess the PPT over the TMJ and (as a reference) the center of the glabella. The mean of the second and third TMJ PPT was used in the analysis, and the ratio between the TMJ PPT and the PPT of the reference site (PPT ratio) was calculated. Temporomandibular joint resting pain and pain upon maximum voluntary mouth opening was assessed by a visual analog scale on each side. RESULTS: The TMJ PPT (median/10th to 90th percentile) and PPT ratio were significantly lower in the RA patients (148/64 to 220 and 0.63/0.40 to 1.01, respectively) than in the healthy individuals (217/111 to 352 and 0.85/0.51 to 1.25), but the overlap was considerable. CONCLUSION: This study shows that the PPT of the TMJ in RA patients is lower than in healthy individuals and that it can be used for pain assessment. However, the clinical use of the TMJ PPT and PPT ratio measurements alone is limited from a diagnostic point of view. PMID- 14737878 TI - A human model of intraoral pain and heat hyperalgesia. AB - AIM: To examine, in a double-blind and placebo-controlled crossover manner, the effect of topical application of capsaicin on the alveolar mucosa with a battery of intraoral quantitative sensory testings (QST) in 16 healthy volunteers. METHODS: Thirty microL of 5 mg/mL capsaicin or vehicle (control) was applied to a 3 x 3-mm paper disk and applied to the alveolar mucosa under an oral bandage. The subjects rated the perceived pain intensity on a 0 to 10 electronic visual analog scale (VAS) for 15 minutes. Quantitative sensory testings were performed before and immediately after the 15-minute application and consisted of assessments of cold detection threshold, warmth detection threshold (WDT), cold pain threshold, beat pain threshold (HPT), mechanical sensitivity to single and repeated punctate mechanical stimulation with von Frey filaments and to single and repeated brush stimulation with a cotton swab, and detection and pain thresholds to electrical stimulation of the alveolar mucosa and maxillary first premolar tooth. Analysis of variance was used to test the data. RESULTS: Application of capsaicin caused moderate levels of pain (VASpeak scores 5.0 +/- 1.9) whereas the vehicle was practically painless (VASpeak 0.9 +/- 2.4). No significant effects of vehicle on QST could be detected (P > .143). In contrast, capsaicin application was associated with significant decreases in WDT and HPT (P < .001). No other significant changes in QST were observed for capsaicin application. CONCLUSION: The intraoral capsaicin pain model is associated with signs of heat hyperalgesia, but not mechanical hyperalgesia. Since the somatosensory sensitivity is not well characterized in most orofacial pain conditions, mainly due to lack of tradition and techniques, intraoral QST may provide a better description of the somatosensory sensitivity and underlying mechanisms in orofacial pain conditions. PMID- 14737879 TI - Self-registration of mandibular opening capacity and vertical overbite. A method study. AB - AIMS: To evaluate a method for self-registration of maximum mandibular opening capacity by means of a spatula and estimation of vertical overbite from photos. METHODS: The study group consisted of 50 adults. Each participant received written instructions, photos, a measuring spatula, and a pencil. The first task was to register maximum interincisal distance. The participant opened up his or her mouth as wide as possible, looked in a mirror, and marked the distance on the spatula. The second task was to estimate the vertical overbite in the intercuspal position. A clinician then estimated the type of overbite and measured the maximum interincisal distance and the vertical overbite with a ruler in millimeters. RESULTS: The normal overbites showed a mean value of 2.4 mm, and a standard value of 2 mm was set. The deep bites showed a mean value of 5.2 mm, and a standard value of 5 mm was set. The standard overbite value, added to the value measured on the spatula, was compared with the clinical values made by the clinician for maximum mandibular opening. The limits of agreement for 95% of the mean difference were -4.2 mm and 4.4 mm. Six people missed the correct diagnosis for the vertical overbite. CONCLUSION: The self-registration method of measuring maximum mandibular opening capacity seems valid for studying major differences in opening capacity when clinical measurements are not possible to perform. PMID- 14737880 TI - Parents' ability to perceive pain experienced by their child with Down syndrome. AB - AIMS: To investigate parents' ability to perceive pain experienced by their offspring with Down syndrome (DS). METHODS: Data were gathered by the use of the Oral Assessment in Down Syndrome Questionnaire in a cross-sectional survey design in France. A sample of parents of 204 children with DS and 161 of their siblings without DS was accrued. RESULTS: Parental reports of difficulty discerning if their child with DS was in pain did not change with age of the child, remaining at a prevalence of 28% to 32%. Reports of difficulty discerning where that child felt pain diminished with older age from 74% to 27%. The likelihood of parents reporting difficulty discerning if and where their child with DS had pain was greater than for a sibling without DS. However, reports of pain experience for the 2 groups were the same. Moreover, different functional and dysfunctional behavioral variables were found to be predictors of these 2 pain perception variables. CONCLUSION: Parental perception of pain is less discriminant for children with DS than for their siblings without DS. PMID- 14737881 TI - European Academy of Craniomandibular Disorders. September 19-20, 2003. Bordeaux, France. PMID- 14737882 TI - Guidelines for curriculum of undergraduate and postgraduate education in orofacial pain and temporomandibular disorders in Europe. PMID- 14737885 TI - [Surgical treatment of pertrochanteric fractures using dynamic methods of internal and external fixation]. AB - Pertrochanteric fractures usually occur in patients over 65 years of age, with greater loss of skeletal mass (osteoporosis). Nonsurgical methods of treatment are accompanied by relatively high lethality rate. Moreover, they do not produce satisfactory anatomical and functional results. Surgical treatment by using dynamic implants represents a method of choice in the fixation of pertrochanteric fractures. This paper presents the treatment results of 110 patients, 61 with pertrochanteric fractures, who were surgically treated by the dynamic method of internal fixation, and 49 patients who were treated by the method of external fixation. Dynamical implants enabled both dynamization and compression of the fracture in the axis of the neck, as well as the diaphysis of the femur, which lowered the risk of mechanical complications, and, at the same time, provided effective healing of the fracture, early activation, and mobilization of the patients on whom the surgery was performed. In patients infected by various diseases, for whom surgical trauma represents a life threat, the external fixation is recommended as a method of choice. PMID- 14737884 TI - Influence of the elevated ambient temperature on immunoglobulin G and immunoglobulin G subclasses in sera of Wistar rats. AB - The aim of our research was to examine changes in the immune system of the rats influenced by the elevated ambient temperature. Male Wistar rats were divided, into 2 groups and housed at 20 +/- 2 degrees C (n = 64, control group) and 35 +/- 1 degrees C (n = 74, experimental group), during precise timing of 1, 4, 7, 14, 21, and 30 days. All the animals were given food and water ad libitum, and were lighted during 12 hours per day. We have measured IgG, IgG1, IgG2a, IgG2b and IgG2c. The obtained results showed significant elevation in the level of IgG after 4 and 7 days (+32%), IgG2a after 7th (+88%), 14th and 21nd day (+110%), IgG2b after 14 days (+60%) at 35 +/- 1 degrees C compared with the control group at 20 +/- 2 degrees C. IgG1 level was not affected and IgG2c showed significant decrease after 21st day at 35 +/- 1 degrees C. In conclusion, during the elevated ambient temperature the immune system is activated as one of the regulation mechanisms in homeostasis and survival of the population. PMID- 14737887 TI - [Diagnostic value of the F-reaction in localization of the level of peripheral motor neuron lesions]. AB - F-wave is a secondary muscle response, which occurs by antidromic activation of the limited number of alphamotoneurons. Based on M- and F-response and the nerve length, the conduction time, conduction velocity, and the ratio of F-response can be determined. These parameters help in assessing the data about functional condition of peripheral motor neurons along the most proximal segment in relatively simple and non-invasive way. In routine practical work, minimal latency of F-wave is measured, while F-wave conduction time, F-wave conduction velocity and F-ratio are not measured. The aim of this research was to examine the importance of F-wave latency, F-wave conduction time, F-wave conduction velocity, and F-ratio in the localization of level of lesion of peripheral motor neurons. Electromyoneurography examination included 50 patients with radiculopathies of lumbosacral part of spinal cord, 50 patients with neuropathy, and 25 healthy persons. Minimal latency of F-wave for tibial nerve and peroneal nerve was determined after 10 stimulations. F-wave latency, F-wave conduction time, F-wave conduction velocity, and F-ratio were determined in accordance with the formulas defined by Kimura. The results of our test have shown that F-wave latency, F wave conduction time, F-wave conduction velocity, were important electrophysiological parameters, while F-ratio was found to be highly sensitive, which made it the most valid electro-physiological parameter in the differentiation of proximal and distal lesion of peripheral motor neuron. PMID- 14737886 TI - [Microvascular free flaps and care of war injuries with tissue defects]. AB - BACKGROUND: War wounds caused by modern infantry weapons or explosive devices are very often associated with the defects of soft and bone tissue. According to their structure, tissue defects can be simple or complex. In accordance with war surgical doctrine, at the Clinic for Plastic Surgery and Burns of the Military Medical Academy, free flaps were used in the treatment of 108 patients with large tissue defects. With the aim of closing war wounds, covering deep structures, or making the preconditions for reconstruction of deep structures, free flaps were applied in primary, delayed, or secondary term. The main criteria for using free flaps were general condition of the wounded, extent, location, and structure of tissue defects. The aim was also to point out the advantages and disadvantages of the application of free flaps in the treatment of war wounds. METHODS: One hundred and eleven microvascular free flaps were applied, both simple and complex, for closing the war wounds with extensive tissue defects. The main criteria for the application of free flaps were: general condition of the wounded, size, localization, and structure of tissue defects. For the extensive defects of the tissue, as well as for severely contaminated wounds latissimus dorsi free flaps were used. For tissue defects of distal parts of the lower extremities, scapular free flaps were preferred. While using free tissue transfer for recompensation of bone defects, free vascularized fibular grafts were applied, and in skin and bone defects complex free osteoseptocutaneous fibular, free osteoseptocutaneous radial forearm, and free skin-bone scapular flaps were used. RESULTS: After free flap transfer 16 (14.4%) revisions were performed, and after 8 unsuccessful revisions another free flaps were utilized in 3 (37.5%) patients, and cross leg flaps in 5 (62.5%) patients. CONCLUSION: The treatment of war wounds with large tissue defects by the application of free microvascular flaps provided shorter wound-closing period, earlier beginning of physical therapy, as well as the treatment of great number of patients with the extensive tissue defects in the conditions of massive influx of the wounded. PMID- 14737888 TI - [Effect of a two-week program of individually monitored physical activity on insulin resistance in obese non-insulin-dependent diabetics]. AB - It is well known that under the influence of regular, individually measured aerobic physical activity, it is possible to raise the biological efficiency of insulin by several mechanisms: by increasing the number of insulin receptors, their sensitivity and efficiency, as well as by increasing glucose transporters GLUT-4 on the level of cell membrane. The aim of this research was to examine whether decreased insulin resistance could be achieved under the influence of the program of individually measured aerobic physical activity in the 2-week period, in the obese type 2 diabetes patients with the increased aerobic capacity (VO2)max. In 10 type 2 diabetes patients 47.6 +/- 4.6 years of age (group E), in the 14-days period, program of aerobic training was applied (10 sessions--35 min session of walking on treadmill, intensity 60.8 +/- 5.7% (VO2)max, frequency 5 times a week), as well as 1,600 kcal diet. At the same time, other 10 type 2 diabetes patients 45.9 +/- 5.5 years of age (group C) were on 1,600 kcal diet. Before and after this period the following was measured in both groups: insulin sensitivity (M/I) by the method of hyperinsulin euglycemic clamp, and (VO2)max by Astrand test on ergocycle. In contrast to the group C, in the second testing of E group subjects a significant increase was obtained in M/I (1.23 +/- 0.78 vs. 2.42 +/- 0.95 mg/kg/min/mU p < 0.001, 96.75%) as well as the increase of (VO2)max (26.34 +/- 4.26 vs. 29.16 +/- 5.01 ml/kg/min p < 0.05, 10.7%). The results had shown that 2-week program of aerobic training had had significant influence on the increased aerobic capacity and insulin sensitivity in the tested patients. PMID- 14737889 TI - [Acute poisoning with cardiovascular agents]. AB - In order to determine the frequency, severity of poisoning, and the efficacy of the applied therapeutic measures, retrospective study of 391 patients treated for acute drug poisoning was performed during one-year period at the Clinic for Emergency and Clinical Toxicology and Pharmacology. In 49 (12.5%) patients cardiovascular agents were the cause of poisoning, most frequently beta-blockers and calcium antagonists (77.5%). Poisoning with antihypertensive agents was registered in 12.2% of patients, antiarrhythmics in 8.2%, and cardiotonics in 2.1%. Beta-blockers and calcium antagonists caused severe poisoning in over 40% of cases. Predominant clinical manifestations were registered on cardiovascular system, while central nervous system effects occurred secondary to cardiotoxicity. Symptomatic and supportive measures were performed most frequently, while specific agents, glucagon, calcium salts, and others, were used less often. PMID- 14737890 TI - [Diet and gastric cancer]. AB - The aim of this case-control study, conducted in Serbia during the period 1998 2000, was to investigate whether diet was associated with the development of gastric cancer. The case group consisted of 131 patients with histologically confirmed gastric cancer, and the control group of 131 patients with orthopedics diseases and injuries. Cases and controls were individually matched by age (+/- 2 years), gender, and place of residence. On the basis of multivariate logistic regression analysis, following factors were found as independent risk factors for gastric cancer: more frequent consumption of high-fat milk [Odds ratio (OR) = 1.45, 95% confidence interval (CI) = 0.99-2.16]; mutton, lamb and/or calf meat (OR = 2.46, 95% CI = 1.11-5.47), sugar (OR = 2.13, 95% CI = 1.43-3.18), semi white bread (OR = 2.09, 95% CI = 1.25-3.50), and salting food (OR = 5.72, 95% CI = 2.63-12.42). Factors found as protective were: more frequent consumption of margarine (OR = 0.41, 95% CI = 0.25-0.69), "other" cheeses (OR = 0.47, 95% CI = 0.29-0.77), and fish (OR = 0.39, 95% CI = 0.19-0.76). PMID- 14737891 TI - [Current treatment of cardiogenic shock]. PMID- 14737892 TI - [Colchicine in dermatology]. PMID- 14737893 TI - [Autologous pluripotent progenitor cells in the treatment of ischemic heart disease]. PMID- 14737894 TI - [Capillary electrophoresis--a new method for analysis of molecules and a challenge for biochemists]. PMID- 14737895 TI - [Management of war burns of the lower leg using free flaps]. AB - Free flaps are used in the surgical treatment of burns for wound closure where the burn is too deep, and in case, when after necrotic tissue excision, the bones, tendons, nerves, and blood vessels remain bare. Covering of the exposed structures is commonly performed in the primary delayed, or in the secondary wound treatment. The possibilities of covering the defects of the lower leg with local flaps are limited. Free flaps are used when all the possibilities of the other reconstructive procedures have been exhausted. The defect of the soft tissue of the lower leg was covered with free flaps in the injured soldiers with deep burns, treated at the Clinic for Plastic Surgery and Burns, Military Medical Academy, Belgrade. In one patient the wound closing was performed immediately after excision of necrotic tissues, and in the other two in the secondary management. The application of free microvascular flaps enabled the closure of large post excision defects of the lower leg in one operation. Our experience in the treatment of these soldiers point to the possibility of coverage of the exposed deep structures with free flaps as early as possible. PMID- 14737896 TI - [Menetrier's disease associated with ulcerative colitis]. AB - In this paper a 21 year old patient was presented with Menetrier's disease, associated with ulcerative colitis. The first symptoms of ulcerative colitis occurred at the age of eleven, since when the patient has been conservatively treated several times because of the exacerbations of the disease. During control examinations presence of polyploid changes in stomach was discovered by upper endoscopy. Gastrectomy was suggested because the patient had excessive anemic syndrome which required weekly substitutional therapy with deplasmatic eritrocytes, as well as hypoproteinemia, while multiple polyploid changes suspect for malignancy were gastroscopically identified. Patient accepted surgical treatment, and was transferred to the Clinic of Surgery. Total gastrectomy was performed, and patohistological finding confirmed Menetrier's disease. After two weeks, the patient was released from the hospital in good general condition, with regular clinical and laboratory findings. PMID- 14737897 TI - [Changes in the oral caused by scurvy]. AB - This report presents a case of a psychiatric patient with scurvy. Upon the clinical examination of the oral cavity, dry and pale lips were noted. Gingiva was highly edematous, soft, purple blue with ulcerated margins. Bleeding was noted upon slightest provocation. Teeth were also affected by the vitamin C deficiency with multiple caries. The patient was asthenic and adynamic. Petechial bleeding and hematomas were present on the skin of extremities. Therapy consisted of rehydration and vitamin C compensation--2 g daily i.v. during 10 days period, and 1.5 g daily orally during the following 2 months. The patient maintained only partial plaque control, and complete recovery of the oral cavity was not established because the patient didn't comply. Concerning that scurvy is a rare disease nowadays, it is very important to recognize its clinical signs, and to establish a valid and prompt diagnosis, because, when untreated, scurvy can have severe consequences on the entire organism. PMID- 14737898 TI - [Serbian military surgical experience (1876-1918) part II: military surgery in Serbia during the Serbian-Bulgarian war]. PMID- 14737899 TI - [DNA: 50 years' of the double helix]. PMID- 14737900 TI - Healthy teeth means better students. PMID- 14737902 TI - A place to go where someone cares. PMID- 14737901 TI - On the frontlines. Fighting the war against abuse from the dental office. PMID- 14737903 TI - Don't call me Bernice. PMID- 14737904 TI - Final summit. Transferring the goodwill of the practice. PMID- 14737905 TI - Taxing problems. California dentist lands in hot water over tax dispute. PMID- 14737906 TI - Election reform proposals run into a landmine in San Francisco. PMID- 14737907 TI - Hygiene, budget issues top ISDS House deliberations. PMID- 14737908 TI - Mind your chairside manners. PMID- 14737910 TI - Bed-wetting, ear infections linked to mouth. PMID- 14737909 TI - Hidden sugars, dental dangers. PMID- 14737911 TI - Lasers gain acceptance. PMID- 14737912 TI - Study: "mother's milk" no longer safe. PMID- 14737913 TI - Recipient of first tongue transplant recovering. PMID- 14737914 TI - Wiping unintended egg off your legislative face. PMID- 14737915 TI - The impact (factor) of biological research for nursing. PMID- 14737916 TI - Emerging genetic technologies in clinical and research settings. AB - With the rapid expansion of genomic health care, nurses are exposed to emerging genetic technologies in a wide variety of clinical and research settings; however, nurses have limited knowledge about these technologies. The polymerase chain reaction procedure, which is the foundation of current molecular genetic technologies, real-time polymerase chain reaction, and microarray analysis are described in this article. The applications, strengths, and limitations of each technology are discussed. PMID- 14737917 TI - Cognitive executive dysfunction in children with mild sleep-disordered breathing. AB - In children, moderate or severe sleep-disordered breathing (SDB) may impair cognitive executive functions (EFs), including working memory, attention, and mental flexibility. The main objective of this study was to assess EFs in children with mild levels of SDB. Subjects for this descriptive study were 12 children (5 girls, 7 boys) aged 8.0 to 11.9 years (M = 9.0 +/- 0.85) participating in an ongoing study of the effects of adenotonsillectomy on behavior. Each subject had a nocturnal polysomnogram (PSG) and a multiple sleep latency test (MSLT). Mild SDB was considered present if the child's apnea/hypopnea index (AHI) was > or = 1 and < 10. Between MSLT nap attempts, each child completed standardized tests of EFs. The sample showed significant impairment of sustained attention and vigilance on a computerized continuous performance test. Children with low mental flexibility scores on the Children's Category Test (CCT) spent more time in stage 1 sleep (12.2% v. 9.5%, P = 0.028 on PSG) and showed a marginally higher arousal index (9.7 v. 6.5, P = 0.06 on PSG) than children with average or above-average CCT scores. AHI accounted for a significant proportion of the variance in CCT scores when 1 outlier was removed (N = 11, Rsq = 0.67, P = 0.002). Mild levels of SDB and associated sleep architecture disruptions may be associated with impairment of EFs in children. PMID- 14737918 TI - Antepartum bed rest: maternal weight change and infant birth weight. AB - Despite lack of evidence for effectiveness, obstetricians in the United States prescribe antepartum bed rest for more than 700,000 women per year. However, in nonpregnant samples, bed rest treatment produces weight loss. This study assessed maternal weight change (gain) during antepartum hospitalization for bed rest treatment; compared appropriateness of infant birth weights for gestational age, race, and gender; and determined whether maternal weight change predicted infant birth weight. The convenience sample for this longitudinal study consisted of 141 women with high-risk pregnancies who were treated with hospital bed rest. Weekly rate of pregnancy weight change by body mass index was compared with Institute of Medicine recommendations for rate of pregnancy weight gain. Infant birth weight was compared with current US infant birth weights for matching gestational age, gender, and race. Weekly antepartum weight change was significantly lower than Institute of Medicine recommendations (P < 0.001). Infant birth weights were also significantly lower than the national mean when matched for each infant's gestational age, race, and gender (P < 0.001). Maternal weight change predicted infant birth weight (P = 0.05). Bed rest treatment is ineffective for improving pregnancy weight gain. Lower infant birth weights across all gestational ages suggest that maternal weight loss during bed rest may be associated with an increased risk of fetal growth restriction. A randomized trial comparing women with high-risk pregnancies who are ambulatory with those on bed rest is needed to determine whether bed rest treatment, underlying maternal-fetal disease, or both influence inadequate maternal weight gain and poor intrauterine growth. PMID- 14737919 TI - Axillary and thoracic skin temperatures poorly comparable to core body temperature circadian rhythm: results from 2 adult populations. AB - Data from 2 separate studies were used to examine the relationships of axillary or thoracic skin temperature to rectal temperature and to determine the phase relationships of the circadian rhythms of these temperatures. In study 1, axillary skin and rectal temperatures were recorded in 19 healthy women, 21 to 36 years of age. In study 2, thoracic skin and rectal temperatures were recorded in 74 healthy women, 39 to 59 years of age. In both studies, temperatures were recorded continuously for 24 h while subjects carried out normal activities. Axillary and thoracic probes were insulated purposely to prevent ambient effects. Cosinor analysis was employed to estimate circadian rhythm mesor, amplitude, and acrophase. In addition, correlations between temperatures at various measurement sites were calculated and agreement determined. The circadian timing of axillary and skin temperature did not closely approximate that of rectal temperature: the mean acrophase (clock time) for study 1 was 18:57 h for axillary temperature and 16:12 h for rectal; for study 2, it was 03:05 h for thoracic and 15:05 h for rectal. Across individual subjects, the correlations of axillary or thoracic temperatures with rectal temperatures were variable. Results do not support the use of either axillary or skin temperature as a substitute for rectal temperature in circadian rhythm research related to adult women. PMID- 14737920 TI - The effects of non-weight bearing on skeletal muscle in older rats: an interrupted bout versus an uninterrupted bout. AB - Age-related changes in skeletal muscle, in combination with bed rest, may result in a poorer rehabilitation potential for an elderly patient. The purpose of this study was to determine the effects of non-weight bearing (hind limb unweighting [HU]) on the soleus and extensor digitorum longus (EDL) in older rats. Two non weight bearing conditions were used: an uninterrupted bout of HU and an interrupted bout of HU. Twenty-one rats were randomly placed into 1 of 3 groups: control, interrupted HU (2 phases of 7 days of HU, separated by a 4-day weight bearing phase) and an uninterrupted HU (18 uninterrupted days of HU). Following non-weight bearing, the soleus and EDL muscles were removed. Fiber type identification was performed by myofibrillar ATPase and cross-sectional area was determined. The findings suggest that any period of non-weight bearing leads to a decrease in muscle wet weight (19%-45%). Both type I and type II fibers of the soleus showed atrophy (decrease in cross-sectional area, 35%-44%) with an uninterrupted bout of non-weight bearing. Only the type II fibers of the soleus showed recovery with an interrupted bout of weight bearing. In the EDL, type II fibers were more affected by an uninterrupted bout of non-weight bearing (15% decrease in fiber size) compared to the type I fibers. EDL type II fibers showed more atrophy with interrupted bouts of non-weight bearing than with a single bout (a 40% compared to a 15% decrease). This study shows that initial weight bearing after an episode of non-weight bearing may be damaging to type II fibers of the EDL. PMID- 14737921 TI - Levels of fatigue compared to levels of cytokines and hemoglobin during pelvic radiotherapy: a pilot study. AB - Cancer-related fatigue (CRF) is a prevalent and distressing symptom experienced by patients during cancer therapy. One proposed mechanism for the development of fatigue is the increased secretion of proinflammatory cytokines and/or the development of anemia. The major purpose of this pilot study was to investigate the levels of fatigue and cytokines during radiation therapy and determine whether there was a correlation between the two. A secondary purpose was to explore the relationships among hemoglobin values, cytokines, and fatigue. Participants included 15 women diagnosed with uterine cancer, who received curative external radiation therapy. Fatigue was assessed by a self-report instrument (Multidimensional Fatigue Inventory [MFI-20]) and hemoglobin and cytokines (Il-1, Il-6, and TNF-alpha) were measured before, during, and after radiotherapy. The degree of fatigue increased during radiotherapy without a significant change in IL-1, IL-6, or TNF-alpha levels. There was no significant correlation between changes in general fatigue and the changes in IL-1 and TNF alpha. There was a significant negative correlation between the change in IL-6 and general fatigue. The hemoglobin levels did decrease significantly during radiotherapy, but there was no significant correlation between general fatigue and hemoglobin after 3 weeks of therapy or after the completion of therapy. In conclusion, pelvic radiotherapy in women with uterine cancer is associated with increased fatigue. There were no significant relationships between anemia or cytokine levels and fatigue. The pathogenesis of fatigue during radiation therapy remains to be elucidated. PMID- 14737922 TI - Linear and nonlinear approaches to the analysis of R-R interval variability. AB - Analysis techniques derived from linear and non-linear dynamics systems theory qualify and quantify physiological signal variability. Both clinicians and researchers use physiological signals in their scopes of practice. The clinician monitors patients with signal-analysis technology, and the researcher analyzes physiological data with signal-analysis techniques. Understanding the theoretical basis for analyzing physiological signals within one's scope of practice ensures proper interpretation of the relationship between physiolgical function and signal variability. This article explains the concepts of linear and nonlinear signal analysis and illustrates these concepts with descriptions of power spectrum analysis and recurrence quantification analysis. This article also briefly describes the relevance of these 2 techniques to R-to-R wave interval (i.e., heart rate variability) signal analysis and demonstrates their application to R-to-R wave interval data obtained from an isolated rat heart model. PMID- 14737923 TI - The association between moderate drinking and heart rate variability in healthy community-dwelling older women. AB - The relationships among moderate alcohol use, autonomic tone, and arrhythmogenesis in older adults have not been adequately studied. Knowledge about these relationships is of increasing importance in light of population aging and recent epidemiological findings that associate moderate alcohol use with decreased rates of coronary artery disease. The purpose of this study was to assess the association between moderate drinking and autonomic tone in older women. Fifty-two Caucasian female participants (age 69 +/- 5.2) were enrolled in the study. Autonomic tone was estimated by time-domain and frequency-domain measures of heart rate variability. Multivariate analysis revealed that alcohol consumption rate in the sample accounted for approximately one third of the 24-h variability in the SDNN and the SDANN, measures of variability cycle lengths of 24-h and more than 5-min, respectively. Significant contributions of alcohol consumption rate to the shorter-term time-domain measures rMSSD and ASDNN, all frequency-domain measures, and HR were not confirmed. However, repeated measures ANOVA revealed that, between the hours of 0000 and 0600, women who drank approximately 0.5 to 3 standard drinks per day had significantly lower [log] HF and [log] LF power compared to abstainers and a tendency toward sympathetic predominance during the evening and nighttime hours. The authors discuss the implications of these findings. PMID- 14737924 TI - Techniques used in high-scoring and low-scoring 'Roche' vaults performed by elite male gymnasts. AB - The 16 highest-scored Roche vaults (G1) performed during the 2000 Olympic Games were compared with those receiving the 16 lowest-scores (G2). A 16-mm motion picture camera operating at 100 Hz recorded the vaults during the competition. The results of t tests (p < .05) indicated G1, compared to G2, had (a) shorter time of board support, greater normalised average upward vertical force and backward horizontal force exerted by the board, greater change in the vertical velocity while on the board, and greater vertical velocity at board take-off, (b) comparable linear and angular motions in pre-flight, (c) smaller backward horizontal impulse exerted by the horse, smaller loss of the horizontal velocity while on the horse, and greater horizontal and vertical velocities at horse take off, (d) greater height and larger horizontal distance of post-flight, (e) higher body mass centre at knee release, and (f) higher mass centre, greater normalised moment of inertia, and smaller vertical velocity at mat touchdown. Therefore, gymnasts and coaches should focus on sprinting the approach; blocking and pushing off the take-off board rapidly and vigorously; departing the board with a large vertical velocity; exerting large downward vertical force and small forward horizontal force from the hand-stand position while on the horse; departing the horse with large horizontal and vertical velocities; and completing the majority of the double salto forward near the peak of trajectory and releasing the knees above the top of the horse to prepare for a controlled landing. PMID- 14737925 TI - Growth in body size affects rotational performance in women's gymnastics. AB - National and state representative female gymnasts (n = 37), aged initially between 10 and 12 years, completed a mixed longitudinal study over 3.3 years, to investigate the effect of body size on gymnastic performance. Subjects were tested at four-monthly intervals on a battery of measures including structural growth, strength and gymnastic performance. The group were divided into 'high growers' and 'low growers' based on height (> 18 cm or < 14 cm/37 months, respectively) and body mass (> 15 kg or < 12 kg/37 months, respectively) for comparative purposes. Development of gymnastic performance was assessed through generic skills (front and back rotations, a twisting jump and a V-sit action) and a vertical jump for maximum height. The results show that the smaller gymnast, with a high strength to mass ratio, has greater potential for performing skills involving whole-body rotations. Larger gymnasts, while able to produce more power and greater angular momentum, could not match the performance of the smaller ones. The magnitude of growth experienced by the gymnast over this period has a varying effect on performance. While some activities were greatly influenced by rapid increases in whole-body moment of inertia (e.g. back rotation), performance on others like the front rotation and vertical jump, appeared partly immune to the physical and mechanical changes associated with growth. PMID- 14737926 TI - Role of joint torques generated in an optimised Yurchenko layout vault. AB - The purpose of the study was to quantify the muscle torques required in the performance of an optimised Yurchenko layout vault based on a five-segment rigid link model and using input data from an elite female gymnast. At impact, the wrist torque trajectory indicated an extension-flexion action while the shoulder was characterised by extension. The approximate 100 Nm (wrist flexor) and 125 Nm (shoulder extensor) respective peak torque magnitudes indicated that the impact action is not passive in nature. The contribution of joint torques to the adjoining segments was apportioned to the relative components namely; centripetal, gravity and net joint torque components. Despite the presence of both large wrist and shoulder joint torques, the net turning effect on the upper limb and hand segments about their centre of mass (CM) was small. The principal role of the upper limb joint torques was therefore to effect the appropriate joint motions and to support the weight of the gymnast. The performance of the optimum vault was primarily the result of the interplay between the centripetal and the net joint torque components at the wrist, hip and shoulder joints. This has implications to the performer in that successful execution of the vault is principally concerned with the ability to create a high angular momentum for horse impact and to then apply an appropriate level of joint torques that will make optimal use of the initial kinetic condition. PMID- 14737927 TI - A kinematic analysis of a judo leg sweep: major outer leg reap--osoto-gari. AB - Twenty male judo players (10 black belt; 10 novice) executed the major outer leg reap, osoto-gari, with maximal effort. Each throw was recorded within the two dimensional sagittal plane using a 60 Hz video camera. Kinematic data that best described the power of the 'tori's' (thrower's) sweeping leg and the velocity of the 'uke's' (recipient's) falling body were analysed using a Peak Performance Technologies Inc. Motus system. Data from black belt and novice groups were statistically analysed using a one-way MANOVA (p = .05). Statistical analysis found only two variables for the 'tori' (peak angular velocity of the trunk; TTRK, and peak angular velocity of the ankle; TANK) to be significantly different. Large TTRK values for the black belt group indicated a proficient ability to create large momentum on the upper body of the 'uke'. Significantly larger differences in trunk rotation velocities of the 'uke' by the black belt group reinforced this notion. These differences were attributed to good upper body to upper body contact or impact, which is considered an important aspect of the 'osoto-gari'. Significant differences in TANK values stressed the importance of executing plantar flexion near sweep contact. The results emphasised the importance of using the sweeping leg in a sequential kinetic link motion rather than as a single rigid segment. PMID- 14737928 TI - A comparison of knee joint laxity among male and female collegiate soccer players and non-athletes. AB - Female athletes are at least twice as likely to sustain an anterior cruciate ligament injury than male athletes. The underlying cause of ACL injury is multifactorial. However, several researchers have identified knee joint laxity as a possible contributing factor. The purpose of this study was to provide a comparison of knee joint laxity between male and female collegiate soccer players and male and female non-athletes. Thirty-nine (19 male, 20 female) apparently healthy, collegiate athletes from the University of Texas at El Paso's 2001-2002 women's intercollegiate soccer team and men's club soccer team, and forty (20 male, 20 female) non-athletic students volunteered to participate. All participants were tested bilaterally using the KT-1000 MEDmetric knee joint ligament arthrometer. Three tests were used to determine anterior laxity: passive displacement, active displacement, and the Lachman test. The mean passive displacement, mean active displacement, and Lachman (only for the left leg) were significantly lower for the athletic group than for the non-athletic group. There were no significant differences found between males and females for the passive and active drawer tests. However, females had significantly more laxity than males in the Lachman test. These findings suggest that strength and conditioning may play a more significant role in knee joint laxity than the sex of the individual. PMID- 14737929 TI - Accuracy of qualitative analysis for assessment of skilled baseball pitching technique. AB - Baseball pitching must be performed with correct technique if injuries are to be avoided and performance maximized. High-speed video analysis is accepted as the most accurate and objective method for evaluation of baseball pitching mechanics. The aim of this research was to develop an equivalent qualitative analysis method for use with standard video equipment. A qualitative analysis protocol (QAP) was developed for 24 kinematic variables identified as important to pitching performance. Twenty male baseball pitchers were videotaped using 60 Hz camcorders, and their technique evaluated using the QAP, by two independent raters. Each pitcher was also assessed using a 6-camera 200 Hz Motion Analysis system (MAS). Four QAP variables (22%) showed significant similarity with MAS results. Inter-rater reliability showed agreement on 33% of QAP variables. It was concluded that a complete and accurate profile of an athlete's pitching mechanics cannot be made using the QAP in its current form, but it is possible such simple forms of biomechanical analysis could yield accurate results before 3-D methods become obligatory. PMID- 14737930 TI - Pole length and ground reaction forces during maximal double poling in skiing. AB - The purpose of the investigation was to study the relationship between thrust phase duration, ground reaction force, velocity increase after pole thrust and pole angles versus pole length during double poling in roller skiing. Seven male regional elite cross-country skiers volunteered as subjects for the study. The subjects performed a maximal double pole thrust on roller skis with each of the three different pole lengths: 'short', self-selected (normal) and 'long'. The short and long poles were 7.5 cm shorter and 7.5 cm longer than the self-selected pole length. The subjects made seven maximal pole thrusts with each pole length, which were randomly selected during 21 trials. For each trial the subjects accelerated from a 1.2 m high downhill slope attaining a speed of 3.92 m.s-1 before making a maximal double pole thrust on a force plate placed at the bottom of the slope. The vertical (F2), anterior-posterior (Fy) and mediolateral (Fx) reaction forces of the left pole were measured by the force plate. The positions of the pole were recorded in 3-D by an opto-electronic system. Thrust phase duration, impulse, mean force, velocity increase after pole thrust and pole angles were calculated from the recorded data. Double poling with long poles produced a significantly larger propulsive anterior-posterior reaction force impulse and velocity increase than normal (p < .05) and short poles (p < .05). This was in spite of a larger mean anterior-posterior reaction force being produced with short poles. Thus, thrust phase duration was a primary factor in determining propulsive anterior-posterior impulse. For the practitioner, the results can be useful in the selection of pole length when the aim is to increase thrust phase duration, anterior-posterior force impulse and velocity. PMID- 14737931 TI - Cycling on rollers: influence of tyre pressure and cross section on power requirements. AB - The resistance against a cyclist while riding on rollers is due mainly to rolling resistance produced by the deformation of the tyre as it rolls against small diameter drums. Resistance is then combined with wheel speed to set power output. The effect of tyre pressure and cross-section on power was investigated by systematically altering the pressure (552 kPa, 690 kPa, and 827 KPa) in a 20c, 23c, 25c, and 28c tyre of the same design while riding at a wheel speed of 45 kph. Average power over 1 minute was measured with a Power Tap Hub (Tune Corporation, Cambridge, Massachusetts, USA) on five occasions. Statistical significance was evaluated at p < 0.05. Power requirements increased significantly with each reduction in tyre pressure for all tyres and pressures except the 25c between 690 and 827 kPa. The 20c tyre required significantly more power from the cyclist at each tested tyre pressure when compared to the other tyres (which were not different from each other). The differences in resistance from tyre size were not observed when ridden on the road. Additionally, a slightly different tyre design from the same manufacturer responded similarly in the 20c, but was significantly different in the 23c size. It was also observed that power requirements increased significantly when both the wheels were ridden on the rollers as compared to just the rear wheel. These results indicate that the power requirements may be significantly altered by the cyclist by adjusting tyre pressure, tyre cross-section size, tyre type, and with the number of wheels contacting the rollers. However, the magnitude of these power requirements may not be suitable for intense workouts of trained cyclists. PMID- 14737932 TI - Technique and timing in women's inward two and one half somersault tuck (405C) and the men's inward two and one half somersault pike (405B) 3 m springboard dives. AB - The purpose of this study was to compare the inward two and one half somersault dive in a tuck position (405C) performed by females (n = 22) and the inward two and one half somersault dive in a pike position (405B) performed by males (n = 24) to determine changes required by females to successfully perform 405B. Key performance variables in inward dives were also compared to those of backward dives. Video data of the dives performed at the 1999 FINA World Diving Cup were captured and digitized to obtain times and postures of the divers at specific events including takeoff and entry. Estimates of flight height and mass normalised work done on the springboard were obtained from flight times. Some females worked the springboard strongly enough to generate sufficient height and rotation to perform 405B. Males performed 405B comfortably because they achieved better height and rotation than the females. A comparison of backward and inward dives revealed that divers are able to attain greater height in backward dives than inward dives. PMID- 14737933 TI - Development and evaluation of a biomechanics concept inventory. AB - To help instructors in evaluating innovations in biomechanics instruction, a standardised test of the key concepts taught in the introductory biomechanics course was developed. The Biomechanics Concept Inventory (BCI) consists of 24 questions that test four prerequisite competencies and eight biomechanics competencies. Three hundred and sixty seven students from ten universities throughout the United States took the test at the beginning and the end of the introductory biomechanics course. Analysis of a sub-sample of the students showed that the BCI was reliable with typical errors in internal consistency and test retest conditions of 1.4 and 2.0 questions, respectively. Mean pre-test scores (8.5 +/- 2.0) significantly (p < 0.0001) improved to 10.5 +/- 3.2 in the posttest (n = 305). Typical biomechanics students could correctly answer half of the prerequisite questions on the pre-test. Instruction resulted in a mean normalised gain (g) of 13.0% of maximum possible improvement that was consistent with research on traditional instruction in introductory physics courses. It was concluded that the BCI could be an effective tool to evaluate the overall effect of pedagogical strategies on student learning of key biomechanical concepts in the introductory biomechanics course. PMID- 14737934 TI - Chemoprevention of lung cancer: from concept to reality. AB - Among all cancer, lung cancer is the leading cause of cancer death in the western world. The poor lung cancer survival figures argue powerfully for new approaches to control this disease such as chemoprevention, that has been defined as the use of agents that reverse, suppress or prevent lung carcinogenesis. Over 80% of lung cancers are attributed to tobacco and carcinogens from cigarette smoke form the unquestionable link between nicotine addiction and lung cancer. Confoundedly epidemiological studies show that not more than 15% of heavy smokers will ultimately develop lung cancer. The fact that 85% of heavy smokers will not develop lung cancer points to differences in susceptibility. Dietary and genetically determined factors seem to play an important role in modulating individual susceptibility and are closely linked to the chemoprevention approach. Developments in tumor biology and chest radiology have given rise to optimism regarding to earlier diagnosis and improved possibilities for the management of preneoplastic- and early invasive lesions. The current article summarizes the chemoprevention efforts in lung cancer during the last two decades and identifies novel strategies for chemoprevention studies in high-risk individuals based on the developments in molecular targeted therapies and developments in response evaluation by using intermediate biomarker endpoints. PMID- 14737935 TI - Epidemiology of viral hepatitis in the Mediterranean basin. AB - The prevalence of viral hepatitis is high and remains a serious public health challenge throughout the world. New molecular biology techniques provided a better understanding of the viruses over the last decades. Novel therapeutic options seem to be promising but preventing measures including donor screening, immunization against hepatitis A virus (HAV) and hepatitis B virus (HBV), universal use of disposable syringes and implementation of better hygienic conditions play a major role in the control of viral hepatitis. The Mediterranean basin has special demographic and socioeconomic features. We reviewed in this article the seroepidemiological features of viral hepatitis in this particular region. Improving general conditions led to a tendency to be infected in older ages with HAV. Hepatitis B and C virus still remain to be the major causes of chronic hepatitis. The seroprevalence of hepatitis D virus, which was once endemic in the Mediterranean region seem to decrease nowadays whereas hepatitis E virus is still prevalent in some areas. Other viruses such as hepatitis G virus (HGV), TT virus (TTV) and SEN virus do not seem to be a major problem and their clinical importance remains to be determined in further studies. PMID- 14737936 TI - IGF-I triple helix gene therapy of rat and human gliomas. AB - PURPOSE: IGF-I anti-gene technology was applied in treatment of rat and human gliomas using IGF-I triple helix approach. MATERIAL AND METHODS: CNS-1 rat glioma cell and primary human glioblastoma cell lines established from surgically removed glioblastomas multiforme were transfected in vitro with IGF-I antisense (pMT-Anti-IGF-I) or IGF-I triple helix (pMT-AG-TH) expression vectors. The transfected cells were examined for immunogenicity (immunocytochemistry and flow cytometry analysis) and apoptosis phenomena (electron microscopy). 3 x 10(6) transfected cells were inoculated subcutaneously either into transgenic Lewis rats or in patients with glioblastoma. The peripheral blood lymphocytes (PBL) derived from "vaccinated" patients were immunophenotyped for the set of CD antigens (CD4, CD8 etc). RESULTS: Using immunocytochemistry and Northern blot techniques, the transfected "antisense" and "triple-helix" cells showed total inhibition of IGF. Transfected cultures were positively stained either for both MHC-I and B7 antigens--60% of cloned lines, or for MHC-I only--40% of cloned lines. Moreover "triple helix" cells as compared to "antisense" cells showed slightly higher expression of MHC-I or B7. Transfected cells also showed the feature of apoptosis in 60%-70% of cells. In in vivo experiments with rats bearing tumors, the injection of "triple helix" cells expressing both MHC-I and B7 interrupted tumor growth in 80% of cases. In contrast, transfected cells expressing only MHC-I stopped development in 30% of tumors. In five patients with surgically resected glioblastoma who were inoculated with "triple helix" cells, PBL showed an increased percentage of CD4 + CD25+ and CD8 + CD11b-cells, following two vaccinations. CONCLUSIONS: The anti-tumor effectiveness of IGF-I anti-gene technology may be related to both MHC-I and B7 expression in cells used for therapy. The IGF-I antigene therapy of human glioblastoma multiforme increases immune response of treated patients. PMID- 14737937 TI - Growth hormone treatment in pituitary insufficiency: selected cases of children with craniopharyngioma and medulloblastoma. AB - PURPOSE: The work concerns the substitution treatment with growth hormone (GH) in hypopituitary children, including cases that occurred in the course of tumor disease, craniopharyngioma (CP) and medulloblastoma (MB). MATERIAL AND METHODS: The studied population concerned 117 children who presented either somatotropic or polyhormonal pituitary insufficiency (the average age was 12.6 years for girls and 13.6 years for boys). The diagnosis of somatotropic pituitary insufficiency (SPI) was based on insulin and clonidin stimulation tests evaluating GH reserve of hypophysis. The computer tomography (CT) and nuclear magnetic resonance (NMR) examinations were carried out before GH substitution in all children. The tumors (four CP cases and one case of MB) were all found in boys and they were treated with surgery and/or radiotherapy. All studied children, including CP and MB operated patients were treated with human GH (hGH)--Genotropin 16 IU, administered in subcutaneous injections. The daily dose was calculated as 0.5 IU/kg/week. RESULTS: The annual increase of children height before GH therapy was about 3.2 cm. In the first year of GH therapy the difference in children growth between the CP/MB group as compared with the rest of patients was less than 1.0 cm: 9.4 and 10.2 cm/year, resp. During the second year of hormone substitution the growth became slower: average values were 8.2 cm and 7.4 cm/year, resp. In CP and MB patients the height increase calculated as SDS values was significant (2.7 and 1.0 resp.). Control NMR examination performed in CP/MB patients treated with surgery with subsequent hGH therapy did not demonstrate any recurrence of tumor. CONCLUSIONS: After two years of hGH therapy the final height of hypopituitary children, including CP patients, nearly reached the values observed in healthy children. GH therapy did not induce a recurrence of neoplasm in CP and MB patients. PMID- 14737938 TI - Serum DNA as a tool for cancer patient management. AB - PURPOSE: Genetic analysis has shown that cell-free circulating DNA in plasma or serum of cancer patients shares similar genetic alterations to those described in the corresponding tumor. One of the most important alterations involved in carcinogenesis is aberrant promoter methylation. The interest in this field has grown due to the implementation of the methylation-specific PCR (MSP) assay. The main objective of this study is to analyze the methylation status of different genes in tumor and serum DNA obtained at the time of surgery in two different tumor models (glioblastoma [GBM] and non-small-cell lung cancer [NSCLC]) and their relationship to clinico-pathological characteristics and response to chemotherapy. MATERIAL AND METHODS: Using MSP assay, we assessed the methylation status of MGMT, RASSF1A, p16, DAPK, TMS-1 in tumor and serum DNA obtained at time of surgery or stereotactic biopsy from 28 GBM patients and from 51 NSCLC patients. RESULTS: In GBM patients, the prevalence of MGMT, p16, DAPK, and RASSF1A promoter methylation was 38.1%, 66.7%, 52.4%, 57.1%, respectively, in glioma tissue, and 39.3%, 53.6%, 34.3%, 50%, respectively, in serum. A high correlation between methylation in tumor and serum (Spearman test p = 0.0001) was observed. In NSCLC patients, RASSF1A, DAPK and TMS-1 were methylated in 34%, 45% and 35% tumors, respectively, and in 34%, 40% and 34% serum, respectively. A good correlation was found between alterations found in tumor and serum (Spearman test p = 0.0001). CONCLUSIONS: The study of serum or plasma DNA has opened new roads for translational research and new strategies for molecular diagnosis. Due to the similarities of alterations found in serum DNA and primary tumor, we can use this tool to calculate the risk of local or distant recurrence and its relationship with survival and its value in patient follow-up to evaluate response to therapy. PMID- 14737939 TI - Importance of nitric oxide (NO) and adenosine in the mechanism of gastric preconditioning induced by short ischemia. AB - PURPOSE: Gastric mucosa subjected to repeated brief episodes of ischemia exhibits an increased resistance to damage caused by a subsequent prolonged ischemic insult and this is called gastric preconditioning. In this study, L-NNA, a non selective NO-synthase inhibitor, and aminoguanidine, a relative inhibitor of inducible NO-synthase (iNOS), were applied prior to short ischemia (occlusion of celiac artery 1-5 times for 5 min) followed by a subsequent exposure to 0.5 h of ischemia and 3 h of reperfusion (I/R). MATERIAL AND METHODS: Male Wistar rats were used in all studies. RESULTS: Short ischemia reduced significantly I/R induced lesions while raising significantly the GBF and luminal NO content. These effects were attenuated by L-NNA and aminoguanidine and restored by addition of L arginine and SNAP to L-NNA and aminoguanidine. Pretreatment of with adenosine (10 mg/kg i.p.) significantly reduced I/R lesions and accompanying fall in the GBF induced by I/R. These protective and hyperemic effects of standard preconditioning and adenosine were significantly attenuated by pretreatment with 8-phenyl theophylline (SPT, 10 mg/kg i.g.), an antagonist of adenosine A1 and A2 receptors. CONCLUSIONS: We conclude that gastric ischemic preconditioning is considered as one of the major protective mechanism in the stomach that involves key vasodilatory mediators such as NO and adenosine. PMID- 14737940 TI - Diethylnitrosamine may induce esophageal dysplasia after local intramural administration. AB - PURPOSE: To assess the possibility of promoting esophageal squamous cell carcinoma after direct administration of diethylnitrosamine (DEN) into the wall of the esophagus. MATERIAL AND METHODS: Adult male Wistar rats weighing 250-300 g were used in the studies. Via laparotomy, solution of DEN (at the volume of 0.1 ml) was injected directly into the esophageal wall. Animals were divided into 3 groups: CONTROL group--injected with saline, DEN1 group--injected with DEN 100 mg, DEN2 group--injected twice with DEN at the dose of 100 mg with 7 days interval (total dose of 200 mg). RESULTS: Microscopic evaluation after 180 days revealed signs of esophagitis in 20% and 30% subjects in DEN1 and DEN2 group respectively. In 30% of animals from DEN1 and 50% animals from DEN2 group, low grade dysplasia was recognized. The difference between DEN2 and control animals was statistically significant with p < 0.03. Neither high-grade dysplasia nor invasive carcinoma were found in both experimental groups. None of the liver specimens showed the evidence of pathology. CONCLUSIONS: These initial results may indicate the possibility of development of premalignant lesions after local administration of carcinogen into esophageal wall. Observed changes were limited exclusively to esophagus which became the "target organ" in this model. PMID- 14737941 TI - The experimental distention of dissected bile duct for the restoration of its continuity in dogs using a device of own construction. AB - PURPOSE: The segmental resection of constricted bile duct and end-to-end biliary anastomosis could be an attractive alternative in the treatment of benign biliary tract stricture. The aim of this study was to restore the anatomical integrity of the hepatic-common bile duct after an artificially produced defect while maintaining the large duodenal papilla, using microsurgical technique. MATERIAL AND METHODS: The experiments were carried out on 25 mongrel dogs. The common bile duct was ligated in all of the animals during laparotomy, as a model of bile duct obstruction in humans. Relaparotomy was performed 3 days after the initial operation. The segment of bile duct, 4 cm in length was resected together with the ligature. The continuous bile flow into the duodenum was assured by a polyvinyl catheter introduced into both ends of dissected bile duct. The proximal end of the hepatic-common bile duct was fixed to a device constructed by us for the distention of the bile duct (DDBD). The anterior part of the device was exteriorized through a separate fistula and fixed to the abdominal wall. The hepatic-common bile duct distention was gradually continued during 18 days, by pulling out the mobile part of the device. After 18 days the device was removed and the distended proximal end of the hepatic-common bile duct was anastomosed end-to-end with its distal end. The sequels of this procedure were observed for up to 6 months. RESULTS: The hepatic-common bile duct was distended 4 cm within 18 days. The histopathological examination has shown partial damage of the duct framework due to the distention and tension. However the patency of the duct was preserved and the recovery of normal structures were observed after the device was removed and anastomosis fashioned. CONCLUSION: This method, developed by us, offers the possibility of restoring the integrity of injured extrahepatic bile ducts, allowing effective treatment of benign biliary strictures. PMID- 14737942 TI - The role of adenosine receptors for pancreatic blood flow in caerulein-induced acute pancreatitis. AB - PURPOSE: The aim of our study was to evaluate the possible influence of adenosine receptors agonists and antagonists on pancreatic blood flow (PBF) and the development of acute pancreatitis (AP) in rats. MATERIAL AND METHODS: Ninety male Wistar rats were subdivided into ten equal groups, nine rats in each. The study was carried out in two stages. In the first one the first group (control) received i.v. saline infusion for 12 hours. The groups 2-5 (the first stage) received i.v. caerulein infusion as in the first group, but with pretreatment with: in the second group--DPCPX (A1 receptor antagonist), in the third group- CGS 21680 (A2 receptor agonist), in the fourth group--ZM 241385 (A2 receptor antagonist), in the fifth group--IB-MECA (A3 receptor agonist). In the second stage the first group received i.v. caerulein infusion at the dose of 5 micrograms/kg/h for 12 hours. The groups 2-5 (the second stage) received i.v. caerulein infusion as in the first group, but with pretreatment with: in the second group--DPCPX (A1 receptor antagonist), in the third group--CGS 21660 (A2 receptor agonist), in the fourth group--ZM 241385 (A2 receptor antagonist), in the fifth group--IB-MECA (A3 receptor agonist). Pancreatic blood flow was measured by laser Doppler flowmetry. Pancreatic inflammation was evaluated by serum alpha-amylase activity, pancreatic weight and histological changes in the pancreatic tissue. RESULTS: We observed a significant attenuation of serum alpha amylase activity increase (19.1 +/- 2.8 kIU/L vs 30.12 +/- 2.64 kIU/L), pancreatic weight (expressed as percentage of rat's body weight--0.85 +/- 0.16% vs 1.25 +/- 0.14%), and improvement of PBF (79.8 +/- 6.1% vs 60.1 +/- 3.6%), a reduced degree of pancreatic tissue damage (oedema, leukocyte infiltration, vacuolisation of acinar cells) in the third group (CGS 21680 + caerulein) compared with the first group in the second stage (only caerulein infusion). Neither agonists nor antagonists exerterd any appreciable effects on measured parameters in healthy rats. CONCLUSIONS: Pretreatment with A2 receptors agonist seems to be protective against the damage to the pancreas during the course of caerulein-induced acute pancreatitis in rats. This effect could be due to improvement of pancreatic blood flow. This finding could have some therapeutic implications. PMID- 14737943 TI - Chymotrypsin-like activity in rat tissues in experimental acute pancreatitis. AB - PURPOSE: Increase in intracellular chymotrypsin activity was reported during acute pancreatitis. Beside chymotrypsin, there are at least two enzymes with chymotrypsin-like activity: proteasome and lysosomal cathepsin A. Until now it is not known whether and to what extent they contribute to increases in chymotrypsin activity in acute pancreatitis. Our aim was to study organ chymotrypsin-like activities during experimental acute pancreatitis. MATERIAL AND METHODS: Rat cerulein model of acute pancreatitis was used. The chymotrypsin-like activities were assessed in pancreas, liver, lung, heart, spleen and kidney using highly selective synthetic substrates of the proteasome and the cathepsin A, at neutral and acidic pH. Determinations after addition of selective inhibitor were also performed. RESULTS: During acute pancreatitis we found in the pancreas an increase only in neutral chymotrypsin-like activity, as compared to the control animals. In other organs neutral chymotrypsin-like activity did not increase, and in kidney it even decreased. There were no changes in acidic chymotrypsin-like activity in any of organs studied. The studies using the inhibitor of the proteasome showed that the neutral chymotrypsin-like activity in the pancreas of the rats with acute pancreatitis should not be attributed to the proteasome activity, but rather to the chymotrypsin. CONCLUSIONS: Our results did not confirm any significant contribution of proteasome or cathepsin A to increased chymotrypsin-like activity in acute pancreatitis. We showed a decrease in neutral chymotrypsin-like activity of proteasome in the kidney, but the significance of this finding remains to be established. PMID- 14737944 TI - Effect of different treatment methods on survival in patients with pancreatic cancer. AB - PURPOSE: To assess the benefit of treatment modalities on the survival in patients with pancreatic cancer. MATERIAL AND METHODS: Eighty-five patients with pancreatic cancer were treated by surgery, radiotherapy, chemotherapy and combined therapy. The data was reviewed retrospectively and the benefit of various treatment methods to the median survival time of the patients was assessed. RESULTS: Median survival time of the patients diagnosed with local disease treated by radical resection and adjuvant treatment was 21.5 months; with radical resection only--12.6 months (p = 0.6). In patients with locally advanced disease and treated by radical resection and adjuvant therapies the median survival time was 12.1 months and by radical resection only 7.7 months (p = 0.6). For patients treated by palliative surgery, chemotherapy and radiotherapy median survival was 8.8 months and by palliative surgery alone 1.8 month (p = 0.015). 1 year actual survival of patients treated with radical resection and adjuvant therapies was 11.7%. For patients with radical resection only--10.5%. 3-year actual survival for the same groups of patients was 3.5% and 2.3%, respectively. 1-year actual survival of patients treated by palliative methods was 2.3%. CONCLUSIONS: Surgery with adjuvant treatment seems to be beneficial for pancreatic adenocarcinoma patients. In locally advanced and metastatic pancreatic cancer palliative chemotherapy or radiotherapy statistically significantly improved survival. PMID- 14737945 TI - Model of single left rat lung transplantation. Relation between surgical experience and outcomes. AB - PURPOSE: The model of unilateral orthotropic left rat lung transplantation is well known and established experimental procedure. The author's personal learning curve of mastery process of this microsurgical procedure is presented. MATERIAL AND METHODS: During 18 months the author has performed 197 single left lung transplantations on the Thoracic Surgery Ward in University Hospital, Berne, Switzerland. There were 147 allogeneic and 50 isogeneic transplantations done. The allogeneic transplantations were carried out from Brown-Norway to Fischer F344 rats whereas isogeneic transplantations were done among Fischer F344 rats solely. Grafted lung was obtained from the intravenously anaesthetised, oxygen ventilated donor. The implantation was carried out through left posterolateral thoracotomy on the gas anaesthetised, respirator ventilated recipient. The anastomoses of the vessels were done using the cuff technique, bronchi were sutured using continuous running over-and-over suture. Recipients were sacrificed on day 5 post-transplant. All recipients were divided into four consecutive groups. Warm ischaemia time and presence of perioperative pure technical complications were observed. RESULTS: We observed time dependent decline of complications number of consecutive recipient groups, respectively 20, 5, 4, 1. The warm ischaemia time in minutes decreased from 35.6 +/- 5.4 in group I through 26.7 +/- 4.4 in group II, 24.8 +/- 2.3 in group III to 22.0 +/- 3.1 in group IV. CONCLUSIONS: Continuous training of the procedure shortens the average warm ischaemia time and reduces the number of complications. This tedious microsurgical procedure is possible to master by the surgeon. PMID- 14737946 TI - Impact of interferon-alpha therapy on the serum level of alpha-fetoprotein in patients with chronic viral hepatitis. AB - PURPOSE: Assessment of the long-term effect of interferon-alpha (IFN-alpha) treatment on the serum level of hepatocarcinogenesis marker--alpha-fetoprotein (AFP), in patients (pts) with chronic viral hepatitis (c.v.h.) type B and C. MATERIAL AND METHODS: Thirty seven pts (21 with HCV and 16 with HBV infection; 20 women, 17 men, aged 24-62) were included in the study. The pts were administered IFN-alpha in the dose of 9-15 MU per week, thrice weekly, for 16 weeks (HBV group) and 24-52 weeks (HCV group). The effectiveness of IFN-alpha treatment was evaluated on the basis of the HBV DNA and HCV RNA level in the blood. The serum AFP values were determined before and 4-7 years after IFN-alpha treatment. RESULTS: The baseline serum AFP level was increased in 26 out of 37 pts (70%) (14/21 from HCV group; 12/16 from HBV group). After the 4-7 years' follow-up it remained elevated only in 2 out of 37 pts (5%). AFP values significantly decreased after IFN-alpha treatment (17.58 +/- 19.09 IU/ml vs 7.95 +/- 21.78 IU/ml; p < 0.05; normal range 0-5 IU/ml) in both HBV and HCV, responder and non responder groups. CONCLUSIONS: IFN-alpha therapy significantly decreases the serum AFP level in patients with chronic viral hepatitis B and C. Its beneficial clinical effects have been observed both in responders and in non-responders. It could diminish the risk of liver carcinogenesis, however further studies are required to elucidate this issue. PMID- 14737947 TI - IL-15 in the culture supernatants of PMN and PBMC and the serum of patients with Lyme disease. AB - PURPOSE: The aim of this study was to estimate the production of interleukin-15 (IL-15) by neutrophils (PMNs) and peripheral blood mononuclear cells (PBMC) confronted with the serum levels of IL-15 in patients with Lyme disease. MATERIAL AND METHODS: PMN and PBMC were isolated from heparinized whole blood of patients. The cells were incubated for 18 hs at 37 degrees C in a humidified incubator with 5% CO2. After 18 hs incubation, supernatant was removed and assessed for IL-15 using ELISA kits. RESULTS: The results obtained showed significant increase in the ability of patient's PMNs and peripheral blood mononuclear cells (PBMC) to release IL-15. Although PBMC produced higher concentrations of IL-15 than PMN, the quantitative dominance of PMN in the peripheral blood suggest a significant role for these cells in the defense reactions controlled by this cytokine. Similar changes in the secretion of IL-15 by PMN and PBMC in patient group may be caused by the same regulatory mechanisms which influence the functional abnormalities of the cells examined. CONCLUSIONS: A change in the ability of PMN and PBMC to release IL-15 may have various implications for the cellular and humoral response to the Borrelia burgdorferi (B.b.) infection in patients with Lyme disease. PMID- 14737948 TI - Concentration of interleukin-6 (IL-6), interleukin-8 (IL-8) and interleukin-10 (IL-10) in blood serum of breast cancer patients. AB - PURPOSE: Interleukins may stimulate cancer cells growth and contribute to locoregional relapse as well as metastasis. Permanent synthesis and release of these cytokines leads to augmentation of their serum concentration that might be utilized as a marker of immunity status and immune system activation in prognosis and monitoring of the course of cancer. MATERIAL AND METHODS: Therefore, in the present study we assessed the concentration of IL-6, IL-8 and IL-10 in blood serum of breast cancer patients to determine whether it correlates with the disease progression. RESULTS: We showed statistically higher serum concentrations of IL-6, IL-8 and IL-10 in breast cancer patients in comparison with healthy women, which also correlated with clinical stage of breast cancer. CONCLUSIONS: The present study indicates that elevated IL-6, IL-8, and IL-10 serum concentration, are strongly associated with breast cancer and correlate with clinical stage of disease. It was feasible that it can be used to diagnose women with breast cancer and to identify patients with a poor prognosis who may benefit from more aggressive management. PMID- 14737949 TI - An evaluation of laryngeal cancer morbidity time trends in Lithuania. AB - PURPOSE: To make assessments of the rates of cases of larynx cancer in Lithuania in the years 1978-2001 as well as possible trends of changes in the future. MATERIAL AND METHODS: The data contained in the Lithuanian Cancer Register for the period 1978-2001 about new cases as well as the data compiled by the Lithuanian Department of Statistics on the average number of population of Lithuania within the same period in the same age groups have been used in the course of the study. The data have been standardized by age using direct method, in accordance with the European standard; a regression analysis of larynx cancer case rates was made. RESULTS: After standardization of data for the period 1978 2001, tendencies of increase have been registered both among men and women: in 1978 the case rate per 100,000 population was 10.73 among men and 0.26 among women, in 2001 the corresponding data were 11.6 among men and 0.7 among women. Throughout the period the investigated case rate for men was higher than for women. An increasing average age of men and women suffering from this disease has been noticed: average age for men is annually increasing by 0.1566 years and for women--0.0602 years. The forecast for men in the year 2006 is 13.88 cases per 100,000 population and 0.54 for women. CONCLUSIONS: The increase of larynx cancer case rates is growing more rapidly among women than among men, and also average age of the patients is increasing. The forecast is that in 2006 the case rate will be growing up, and both men and women will get ill at older age. PMID- 14737950 TI - Cytokine and adhesion molecule concentrations in blood of patients with B-cell chronic lymphocytic leukaemia with regard to disease progression. AB - PURPOSE: To examine concentrations of IL-8, E-selectin and VCAM-1 in blood plasma, culture supernatant and isolated broken lymphocytes in patients with chronic lymphocytic leukaemia (CLL), at I and III stage of the disease according to Rai's classification and in healthy individuals constituting controls. Two types of lymphocyte cultures were carried out--nonstimulated and stimulated with mitogen. MATERIAL AND METHODS: A concentration assay of substances mentioned above was made using ready immuno-enzymatic sets of the ELISA type. RESULTS: In all cases, plasma concentrations of IL-8, E-selectin and VCAM-1 were the highest in III stage of CLL and moderately increased in I stage comparing to control. The concentrations of IL-8 in culture supernatants and in broken lymphocytes and concentration of VCAM-1 in lymphocytes were increased in both stages of CLL. In contrast concentrations of E-selectin were decreased in lymphocytes in both types of culture. Significant decrease of E-selectin and VCAM-1 concentrations were observed in supernatants of nonstimulated cultures. CONCLUSION: Significant increase of E-selectin, IL-8 and VCAM-1 concentration in blood plasma may support higher activity and suggest the higher ability of leukaemic lymphocytes to migration. Increased IL-8 concentration in isolated, broken, stimulated lymphocytes may be a characteristic feature of the biochemical processes of leukaemic lymphocytes. The lowest values of E-selectin and VCAM-1 concentrations in culture supernatants and broken lymphocytes may suggest their degradation during in culture. PMID- 14737951 TI - Peripheral blood lymphocyte population in children infected with Helicobacter pylori. AB - PURPOSE: Helicobacter pylori infection in children is associated with a chronic inflammatory process of gastric and duodenal mucosa, which may have a various clinical course ranging from asymptomatic and chronic inflammatory condition to gastric ulceration. The immune system may contribute especially to chronic gastric mucosa inflammation. The aim of our study was to assess the levels of peripheral blood T (CD3+, CD4+, CD8+) and B lymphocyte subpopulation (CD19+) in children with Helicobacter pylori infection and to evaluate their relation to degree of antrum mucosa inflammation. MATERIAL AND METHODS: The study was performed in 32 children aged 7-18 years, hospitalized due to dyspeptic symptoms. The endoscopic examination of upper gastrointestinal tract was performed and gastric and duodenal mucosa was estimated in all patients. The endoscopic and histological evaluation of gastric mucosa was performed according to the Sydney System [4]. The urease test (CLO-test-H. pylori) was made to estimate the severity of the infection. RESULTS: Moderate antrum mucosa inflammation was found in 41.2% of the examined. The highest percentage of children (58.8%) presented marked inflammation. No mild inflammation was found in children examined. CONCLUSIONS: No correlation was found between lymphocyte levels and the degree of the inflammatory changes in antrum mucosa. The evaluation of peripheral blood lymphocytes performed in children with Helicobacter pylori infection suggests that T lymphocytes may play a predominant role in this infection. PMID- 14737952 TI - Comparative evaluation of gastric mucosa morphological changes in children and adults with positive IgG antibodies to Helicobacter pylori. AB - PURPOSE: Helicobacter pylori colonization of gastric epithelium causes a local and systemic, cellular and humoral immune response. Despite this immune response involvement in the infection, its elimination from the organism does not take place and the process usually becomes chronic. The purpose of the study was to establish the prevalence of gastric mucosa inflammation in children and adults with serum positive anti-Helicobacter pylori antibodies IgG. MATERIAL AND METHODS: The study included 171 patients comprising 109 (63.7%) children and 62 (36.3%) adults with IgG positive titre against Helicobacter pylori, who were qualified to the study basing on epidemiological examinations estimating the prevalence of Helicobacter pylori infection in the population of north-eastern Poland living in the country, town and city. All patients reported dyspeptic symptoms. The evaluation was performed basing on the morphological (endoscopy) and histopathological examinations estimating the changes in gastric mucosa of these patients. RESULTS: The evaluation of antrum and corpus gastric mucosa proved normal gastric mucosa in 34 children (31.1%) and 10 adults (16.1%) with positive IgG antibodies against Helicobacter pylori. The evaluation of the severity showed the predominance of moderate inflammation within corpus in children (37.6%) and marked inflammation in adults (45.1%). CONCLUSIONS: The concentration of IgG antibodies against Helicobacter pylori in both groups was highest in patients with marked antrum gastric mucosa inflammation. PMID- 14737953 TI - The incidence of some civilization diseases in families of children with food allergy/intolerance. AB - PURPOSE: The aim of the study was to evaluate the incidence of chosen civilization diseases in families of children with food allergy/intolerance. We also wanted to indicate the need for developing and implementing activities preventing these diseases among children. MATERIAL AND METHODS: On the basis of information from questionnaires, two groups of children were distinguished: a group of 80 children suffering from food allergy/intolerance on elimination diet (GR1) and a group of 67 healthy children (GR2) on regular diet. In GR1, the elimination diet with soya bean preparations or casein hydrolysates was introduced before the age of 6 months and continued for at least 12 months. A high risk of hypercholesterolemia according to extended American Academy of Pediatrics criteria including hypertension, diabetes and obesity was determined for children in both groups. RESULTS: The research showed that 31.25% of children examined according to AAP criteria and 46.25% according to extended criteria had a positive family history of premature diseases of the circulatory system. The study proved that hypertension was the most frequent cause of morbidity in families of children from a high risk group and it was found in 67.7% of families with children on elimination diet and with a positive family history and in 78.7% of families with children from GR2 with a positive family history. Obesity, coronary heart disease, hypercholesterolemia, atherosclerosis and diabetes were listed consecutively. CONCLUSIONS: Once a positive family history of cardiovascular diseases is discovered, systematic education promoting health in a family and complex evaluation of physical and psychomotor development of the children should follow. Arterial blood pressure and lipid profile in serum ought to be monitored to eliminate risk factors of these diseases for children. PMID- 14737954 TI - The usefulness of testicular atrophy index in the assessment of undescended testicle--preliminary report. AB - PURPOSE: Cryptorchidism affects 2-8% of male newborns. There is a controversy regarding timing of surgery as well as indications for orchiopexy in boys with retractile testicle. The aim of this study was to evaluate the clinical usefulness of testicular atrophy index (TAI) as a criterion of qualifying patients with undescended testes for surgery as well as of monitoring the results of treatment. MATERIAL AND METHODS: In 1999-2000, 105 cryptorchid boys, aged 1 to 15 years (mean 4.8) underwent unilateral orchiopexy. Dimensions and volume of testes were measured by means of scrotal US and TAI was calculated before and 1 year after surgery. RESULTS: Pre- and postoperative scrotal US measurements were analyzed in 35 boys divided into five age dependent groups. The preoperative TAI values ranged from 27.1% to 52.8%. The biggest loss in volume of affected testis was found in boys aged 4 to 10 years (35.4% to 52.8%). The TAI values measured one year after orchiopexy were lower than preoperative ones. Significant difference in TAI values, ranging from 18.16% to 36.43% were observed in boys between 2 and 10 years (p < 0.001). In the youngest (0-2 yrs) and the oldest boys (> 10 yrs) the difference was not statistically significant. CONCLUSIONS: The testicular atrophy index (TAI) proved to be a valuable and objective tool for qualifying patients with undescended testes for surgery as well as for monitoring the results of treatment. Its value of 20% and more should be considered an indication for surgery in boys with retractile testes. PMID- 14737955 TI - Oxyuriasis-induced intestinal obstruction in a child--case report. AB - PURPOSE: The presentation of an unusual case of the tumor of ileum wall induced by pinworm infection in a 5-years-old child. MATERIAL AND METHODS: The record of a 4-years-old boy treated in the department of pediatric surgery was analyzed concerning the diagnostic difficulties. After 6 month from an episode of ileo cecal intusussception successfully treated with a barium colon enema, the diagnosis of lymphoma was made and the resection of distant segment of small intestine was performed. RESULTS: No clinical and laboratory features of oxyuriasis could be stated before the onset of disease, during hospitalization and in the follow-up period. The hypertrophied and activated lymphatic tissue with a non-specific inflammatory reaction to the pinworms were seen in the wall of ileum, appendix and mesenteric lymph nodes. No neoplastic cells were found in the microscopic study of ileum, appendix, mesenteric lymph nodes and peritoneal lavage fluid. CONCLUSIONS: The proper diagnosis of oxyuriasis may be difficult when the course is atypical. The enterobius vermicularis infestation as an etiologic factor should be taken into account in any case of abdominal pathology. However, the methods routinely used in "acute abdomen" including examinations of blood, urine and stool, repeated ultrasound and CT, are not reliable. As the infestation may mimic neoplasm, the surgical treatment and microscopic examination can be necessary for the final diagnosis in some cases. PMID- 14737956 TI - High cholesterol in patients with ECG signs of no-reflow after myocardial infarction. AB - PURPOSE: Despite successful restoration of blood flow in epicardial artery after myocardial infarction (MI), some patients do not benefit sufficiently from modern revascularisation methods due to the impairment of microcirculation, also called no-reflow phenomenon. Hyperlipidaemia is well established risk factor of coronary heart disease and its detrimental actions on vessels are widely acknowledged. We attempted to investigate possible relations between hyperlipidaemia and electrocardiographic signs of no-reflow in myocardial infarction after successful primary angioplasty. MATERIAL AND METHODS: A total of 150 consecutive patients with acute myocardial infarction (AMI) with ST elevation who underwent successful primary angioplasty were studied. ECG was obtained directly before and 30 minutes after successful reperfusion. ST segment deviation was measured. Lack of 50% reduction of ST-segment elevation in the lead with maximal initial elevation, 30 minutes after angioplasty was defined as ECG sign of no-reflow. RESULTS: ST segment resolution occurred in 116 patients (77%), whereas 34 presented ECG signs of no-reflow (23%). Patients with persistent ST-segment elevation had higher blood LDL and total cholesterol (TC) levels than group with ST-segment restoration (146.5 vs. 128.7 p < 0.01 and 219.5 vs. 200.9, p < 0.05 respectively). Triglyceride, HDL, glucose on admission and fasting glucose levels did not differ significantly between groups. ECG signs of no-reflow were observed more often in patients with anterior AMI, history of prior myocardial infarction and longer pain-to-balloon time (p < 0.05). CONCLUSIONS: Positive relation between impaired tissue perfusion and high TC and LDL blood levels suggests that lipids may play a role in the pathogenesis of no-reflow phenomenon, possibly by impairment of endothelial function. PMID- 14737957 TI - Evaluation of skin barrier function in allergic contact dermatitis and atopic dermatitis using method of the continuous TEWL measurement. AB - PURPOSE: The aim of study was to determine usefulness of the method of continuous TEWL measurement in the evaluation of skin barrier function in physiological conditions and in allergic contact dermatitis (ACD) and atopic dermatitis (AD). MATERIAL AND METHODS: Study was conducted on a group of 86 persons: 48 patients with allergic contact dermatitis, 18 with atopic dermatitis and 20 healthy individuals. Measurements of transepidermal water loss were made using custom constructed device for continuous TEWL measurement. In each person the measurements of TEWL were made 4 times: measurement 0 (baseline)--before occlusion with 1% lauryl sulphate for 24 h, measurement 1-15 minutes after SLS patch removal, measurement 2-30 minutes after measurement 1 and measurement 3-30 minutes after measurement 2. Obtained data were statistically analyzed. RESULTS: TEWL ratio values obtained in measurement 0 were as follows in individual groups of patients: 13.20 +/- 8.25 in the AD patients, 10.09 +/- 8.29 in ACD patients and 9.02 +/- 5.99 in control group. Analogous TEWL values in the subsequent measurements were: in measurement 1--16.08 +/- 11.17; 11.63 +/- 6.43; 17.39 +/- 12.41, in measurement 2--23.72 +/- 14.58; 14.71 +/- 6.46; 17.55 +/- 8.25, measurement 3--24.09 +/- 14.93; 16.34 +/- 6.32; 18.44 +/- 8.26. TEWL ratio values were higher in both groups of patients as compared to control group but not statistically significant (p = 0.1778). After 24 h exposition to SLS, TEWL ratio values increased in all examined groups as compared to baseline (0) measurement. All measurements, except for measurement No 1 in AD group of patients, showed statistically significant differences. The highest increase of TEWL values were observed in group of AD patients. CONCLUSIONS: Delay in skin reaction to SLS in patients with atopic dermatitis provides evidence for different properties of water barrier of the skin in this group as compared to healthy individuals. Increasing tendency in TEWL values 1 hour after SLS removal might reflect persistent damage to water barrier of the skin by detergent. Method of continuous assessment of water barrier of epidermis, through the possibility of multiple measurement by TEWL in examined periods of time, decreased the risk of mistake and increased accuracy of measurement. Measurement of TEWL values allows for assessment of otherwise unnoticed damage to water barrier of the skin. PMID- 14737958 TI - Enhanced release of platelet factor 4 into the circulation in patients with atopic eczema/dermatitis syndrome. AB - PURPOSE: The active participation of platelet in IgE-mediated inflammatory response is well documented. Platelet factor 4 (PF4), a platelet-specific protein may play an important role in the development of the atopic eczema/dermatitis syndrome (AEDS). OBJECTIVE: The aim of the study was to evaluate the activation state of circulating platelets in patients suffering from AEDS. MATERIAL AND METHODS: In vivo platelet activity was assessed by measuring plasma level of PF4 (enzyme-linked immunoassay method) in 9 males AEDS patients and 11 healthy, nonatopic subjects. RESULTS: Plasma PF4 was significantly increased (p < 0.05) in AEDS (31.88 +/- 20.48 IU/ml) patients compared with control subjects (2.95 +/- 0.6 IU/ml). CONCLUSIONS: This result suggests that patients with AEDS may have increased in vivo platelet activation expressed by PF4 release. The aim of the study was to evaluate the activation state of circulating platelets in patients suffering from AEDS. PMID- 14737959 TI - Incidence of elevated LH/FSH ratio in polycystic ovary syndrome women with normo- and hyperinsulinemia. AB - PURPOSE: The aim of this study was to determine the incidence of abnormal LH/FSH ratio in women with polycystic ovary with normo- and hyperinsulinemia and to assess the influence of elevated LH/FSH ratio on selected endocrine and biochemical parameters. MATERIAL AND METHODS: One hundred nineteen polycystic ovary syndrome women in reproductive age hospitalized between 1996 and 2000 in Division of Infertility and Reproductive Endocrinology at Poznan University of Medical Sciences were selected for the study. In all selected women LH and FSH serum levels were determined and LH/FHS ratio was calculated. These groups became the subject of a detailed clinical, hormonal and metabolic analysis, which was performed between 6th and 10th day of a natural or induced menstrual period. RESULTS: LH/FSH ratio greater than 2 was accepted as abnormal, and it was found in 54 women (45.4%; I group). Normal gonadotropin ratio was detected in 65 women (55%; group II). Statistically significant differences were noted between groups with normal and elevated LH/FSH ratio in the following parameters: BMI (body mass index), serum insulin, and LH levels. Further analysis revealed that the majority of women with elevated insulin concentrations belong to the group with normal LH/FSH ratio. CONCLUSIONS: LH/FSH ratio is not a characteristic attribute of all PCOS women: in the present study this abnormality was detected in a subpopulation smaller than 50%. Most of the PCOS women with normal gonadotropin ratio belong to a group of patients suffering from hyperinsulinemia and obesity. Patients with hyperinsulinemia and excess of LH constitute a selected and distinct subgroup with increased adrenal androgenic activity. PMID- 14737960 TI - Uterine contractility signals--an introduction to wavelet analysis. AB - PURPOSE: Analysis of the uterine contractility in the nonpregnant states has provided information about physiological changes during menstrual cycle. There is need to develop methods of recording uterine activity as well as mathematical interpretation of recorded time series. Wavelets are a new powerful tool for signal and image processing. The aim of this study is an introductory view of Fourier (one of the fundamental methods of investigating of biomedical signals) and wavelet transforms applications in the analysis of uterine contractions. MATERIAL AND METHODS: Spontaneous uterine activity of healthy patient and patient with dysmenorrhea was recorded by micro-tip two sensors catheter (Millar Instruments, Inc. USA). After amplification analogue signals were converted to digital. Signals were analysed using Fourier and wavelet transforms. RESULTS: Contrary to the Fourier decomposition, which is global and provides the information integrated over the whole signal, the continuous and discrete wavelet transforms allow to extract local and global variations of the recorded contractions. From the analysis of the coefficients of the wavelet transform we can assess various pattern of propagation: normal propagation, simultaneous propagation and inverted propagation. CONCLUSIONS: This study is the introduction to the wavelet analysis of the uterine contraction signals. Wavelet transform provides insight into the structure of the time series at various scales. It allows to localise changes of the signal in time, providing additional information in comparison with the Fourier transform. PMID- 14737961 TI - The evaluation of dentition status in HIV-infected patients. AB - PURPOSE: The aim of the study was the evaluation of dentition status in patients infected with HIV. MATERIAL AND METHODS: We examined 30 HIV + patients, aged 20 46 and 30 non-infected subjects as the control group. Oral hygiene and dentition status were estimated. Oral hygiene status using simplified OHI-plague index according to Green and Vermilion. Dentition status was analysed using decay intensity index (DMF) as well as teeth loss index according to Rogowiec. The results were analysed in dependence on HIV infection with regard to infection time. RESULTS: The results point to a high intensity of decay in HIV+ patients (23.66). There was a positive correlation between infection time and decay intensity and teeth loss evaluated using Rogowiec index. Unsatisfactory oral hygiene status (OHI > or = 1) was observed in 53.33% of infected patients. There is a relation between infection time and oral hygiene status. OHI-plague index increased in patients with infection time longer than 5 years up to 2.99 (patients with shorter than 5 years infection time--1.17 and the control group- 0.57). CONCLUSIONS: 1. There is a positive correlation between HIV infection and dentition status and oral hygiene. 2. Infection time influences index values: decay intensity, teeth missing, and oral hygiene. 3. HIV+ subjects are patients of high necessity of therapy and because of their basic disease they should come within broadened health education and prophylactic activities. PMID- 14737962 TI - Release of hydroxyl ions from calcium hydroxide preparations used in endodontic treatment. AB - PURPOSE: The purpose of our study was to compare the in vitro release of hydroxyl ions from several calcium hydroxide preparations used in endodontic treatment. MATERIAL AND METHODS: Equal quantities of the materials--nonsetting (pure calcium hydroxide, Biopulp, Calcicure), setting--canal sealers (Sealapex, Apexit) and points were placed in dialysis tubes which were then immersed in deionized water. The release of hydroxyl ions from the preparations was measured by the median pH of the deionized water used for dialysis, by means of a pH-meter. The results of our study were analyzed by means of Tukey's reasonable correlation. Significance difference (one-way variance analysis ANOVA) and Pearson's linear correlation coefficient (r2). RESULTS: Nonsetting preparations of calcium hydroxide have a significantly higher capability of hydroxyl ions release in comparison with sealers and points, irrespective of time (p < 0.05). Sealapex and "plus" points released hydroxyl ions to a much greater extent than both Apexit and "regular" points at most periods of the experiment (p < 0.05). Apexit released significantly more of hydroxyl ions than "regular" points, and Sealapex more than "plus" points in the later periods of the experiment (p < 0.05). The pH values of dialysis samples of all materials correlated positively with time and the pH. Almost all materials reached a maximum on the 8-th day of the experiment. CONCLUSIONS: To achieve maximum concentration of hydroxyl ions in tissues: for temporary root fillings nonsetting preparations of calcium hydroxide should be chosen rather than points and they should be placed for at least one week, for permanent root fillings it is more recommended to use Sealapex than Apexit as a sealer. PMID- 14737963 TI - Effects of ebelactone B on cathepsin A activity in intact platelets and on platelet activation. AB - PURPOSE: Previous in vitro studies have demonstrated that a potent antihypertensive agent ebelactone B inhibits cathepsin A/deamidase activity. The aim of our studies was to assess the effects of this inhibitor on cathepsin A activity in intact platelets and on platelet activation events. MATERIAL AND METHODS: PRP or washed human platelets from healthy volunteers were pre-incubated with different concentrations of ebelactone B (1-10 microM) for 10-60 min. Cathepsin A activity in platelets was assayed colorimetrically using Cbz-Phe-Ala at pH 5.5. Expression of platelet activation markers GpIIb/IIIa and P-selectin on non-activated or agonist-activated platelets (ADP, TRAP) was measured by flow cytometry. RESULTS: Pre-treatment of platelets for up to 60 minutes with 10 mumol/l ebelactone B, that effectively inhibits cathepsin A activity in platelet lysate, did not affect this activity in intact platelets. Exposure of PRP to 10 mumol/l ebelactone B alone, or before platelet activation with ADP or TRAP caused only a small but non-significant increase in P-selectin and GpIIb/IIIa expression on the platelet surface, as demonstrated by flow cytometry analysis. CONCLUSIONS: The lack of cathepsin A inhibition by ebelactone B in intact platelets indicates that this inhibitor does not enter cells. Therefore, a potential antihypertensive significance of this compound may be through the inhibition of cathepsin A/deamidase released from activated or damaged cells. In vitro ebelactone B seems to exert no effect on platelet activation. Further studies are underway to determine whether ebelactone B administration affects platelet activation events in experimental model of hypertension in rats. PMID- 14737964 TI - The pattern--reversal visual evoked potentials in children with migraine with aura and without aura. AB - PURPOSE: Studies of the visual evoked potentials (VEP) in migraine have yielded contradictory results. Several investigators suggested that VEP may be helpful test in diagnosis of a child with headache. The aim of our study was to compare interictal pattern-reversal visual evoked potentials (PR-VEP) in children and adolescents with migraine and tension-type headaches and to evaluate VEP parameters in migraine with and without aura. MATERIAL AND METHODS: The study was carried out in 93 children and adolescents aged 7-18 years with attack headaches. RESULTS: 51 children had diagnosed migraine. In this group 30 children (59%) had migraine without aura (MO), 12 children (23.5%) migraine with aura (MA) and 9 (17.5%) patients other variants of migraine (MV): hemiplegic, ophthalmoplegic, basilar. In control group 42 children were classified as tension-type headaches. All children had PR-VEP performed in headache-free period, without prophylactic treatment. The P100 mean latency was significantly longer in migraine than in tension-type headache. Amplitudes N1-P100 and P100-N2 were significantly larger in migraneurs compared with tension-type headache. The mean amplitudes of N1-P100 and P100-N2 were significantly lower in MA compared with group MO. There were no statistically significant differences of other PR-VEP parameters between MA, MO, MV. If we compare individual results of each patient with migraine with mean value +/- 2 standard deviations (SD) of tension-type headaches group, only 25% have VEP abnormalities of latency or amplitude above 2SD value in tension-type headache group. CONCLUSIONS: The diagnosis of migraine in children actually remains predominantly based on medical history, due to low sensitivity and specificity of electrodiagnostic tests in headaches. However PR-VEP may support the diagnosis of migraine in some cases. VEP could be also helpful method in studying the pathogenesis of different forms of migraine. VEP abnormalities in migraine can be related to a cortical spreading depression and a central neurotransmitter alterations. PMID- 14737965 TI - [Glycine as an inherent component of bacterial lipopolysaccharides]. AB - The amino-acylation of bacterial polysaccharide antigens and the biological role of this phenomenon are poorly understood, although it might be relevant in the processes of the infection and immunity. Due to the lability of ester linked substituents on glyco-conjugate antigens, usually such groups escaped detection during routine structural investigation. Among the only few data available, these on the occurrence of glycine in endotoxic lipopolysaccharides of Gram-negative bacteria are well documented. This work summarises these data on glycine as an integral constituent of bacterial LPS and also on some other amino acid esters in teichoic acids and phosphatidylglycerol of Gram-positive bacteria. The possible functions are discussed of such noncarbohydrate ester linked substituents in bacterial antigens. PMID- 14737966 TI - [The role of GABA-ergic system in carcinogenesis]. AB - The gamma-aminobutyric acid (GABA) is the major and the best known neurotransmitter with inhibitory properties in the central nervous system (CNS). Outside the CNS, GABA acts as a regulator of muscle tension and controls the secretion of hormones. In the peripheral region, as in the CNS GABA's function is known to be mediated by GABAA and GABAB receptors. Several recent reports have suggested a relationship between the GABA-ergic system and oncogenesis. It has been confirmed that both GABA content and GAD activity are increased in material from colon, breast, digestive tract, and ovarian cancer. In the light of theory of dynamic balance between stimulating and inhibitory amino acids, disturbances in GABA metabolism may be a sign of the cell's defensive reaction during carcinogenesis. PMID- 14737967 TI - [Dendritic cells and their applications in cancer immunotherapy--achievements and future prospects]. AB - Dendritic cells (DC) are generally believed to play a key role in the initiation of immune response. A potential usefulness of these cells in antitumor immunotherapy is strongly considered in every stage of cancer treatment. Studies on tumor tissue infiltration by immune cells shown, that DC represented only a small percentage of leukocytes. The influence of tumor environment resulted in reduction of DC number, their inability to migrate across endothelial barriers, or impaired maturation and efficiency of tumor antigens presentation. Thus, decreased number of DC in tumors could be associated with a bad prognosis. Many attempts concentrate on the creation of the DC-based vaccines, which would generate strong anticancer cell mediated immunity. They include studies on stage of differentiation of the administered DC; effective way for antigen loading; optimum route and schedule of DC delivery into tumor bearing host; effective vectors for the therapeutic genes; effects of the therapy based on cytokine secreting DC; influence of DC on co-operation between innate and acquired immunity as well as on the generation of specific antitumor response. This review is focused on two important areas aiming for the preparation of DC vaccine for effective stimulation of immune response: loading of DC with tumor antigens (or other ways of DC preparation to successful antigen presentation)--as an encouraging evidence of therapeutic efficacy, and genetic modification of DC with cytokines resulting in the stimulation or alteration of the antitumor DC activity -as a promising anticancer strategy. PMID- 14737968 TI - [Structure and function of lymphocyte TCR/CD3 complex]. AB - The multi-chain T cell receptor/CD3 complex (TCR/CD3) plays a key role in antigen recognition, T cell activation and in consequence in triggering an antigen specific immune response. This process is induced by direct interaction of the TCR receptor with an antigen bound to the major histo-compatible complex on antigen-presenting cells. Upon the structural and functional cooperation of TCR receptor with CD3 complex, the activating signal is transmitted through the cell membrane to the nucleus. The pivotal role in signaling cascade plays CD3-zeta (zeta) chain, which triggers many biochemical events and second messenger activation, leading to the transcriptional factors expression and further T cell proliferation, effector function augmentation and cytokine production. PMID- 14737969 TI - [The pathogenesis and the adaptive value of fever]. AB - Fever is a part of the acute phase response to infection and inflammation. We now understand that fever is a complex physiological response that is aimed at facilitating survival of the host. The fever is induced by endogenous inflammatory mediators, such as prostaglandins and pyrogenic cytokines, that are released by immune cells activated by exogenous pyrogens. Although the pathways (humoral and/or neuronal) responsible for transfer of the pyretic signals from the blood to the brain are still under discussion, it is generally accepted that they act on the level of the anterior hypothalamus to raise the thermoregulatory set-point. Results of studies of the adaptive value of fever demonstrate an association between a rise in body temperature and a decrease in mortality and morbidity during infection. These data along with data from evolutionary studies provide a strong support for the concept that fever is a beneficial during infection in endotherms and ectotherms, vertebrates as well as in invertebrates. There are also evidence showing that fever may be used as a therapeutic tool, especially in cancer therapy. Based on the data reviewed in this article, it can be concluded that fever has evolved as a host defense mechanism which was preserved within the animal kingdom through hundreds of millions of years of evolution. PMID- 14737970 TI - [Methods of collagenous tissue fixation in the preparation of bioprostheses]. AB - The use of biological materials in construction of bioprostheses requires the application of different chemical or physical procedures of fixation increasing bioprostheses resistance to enzymatic or chemical degradation and reducing their antigenicity. Methods typically concentrate on creating additional intra- and intermolecular chemical bonds between collagen molecules. This review focuses on the various methods of stabilization of collagenous tissues including chemical fixatives and physical agents. PMID- 14737971 TI - [Role of adiponectin--a protein secreted by adipose tissue in preventing atherosclerosis]. AB - Adiponectin is an adipocyte-specific secretory protein, which seems to play a protective role in different models of vascular injury. Adiponectin infiltrates in the subendothelial space of injured vascular walls and suppresses the expression of adhesion molecules on endothelial cells, thus inhibiting the inflammatory processes that occur during the early phases of atherosclerosis. Adiponectin also suppresses lipid accumulation in macrophages and macrophage-to foam cell transformation. The ability of adiponectin to act as an anti inflammatory and anti-atherogenic factor has made this novel adipocytokine a promising therapeutic tool for the future. PMID- 14737973 TI - [Fresh blood! A mandatory support for nephrology]. PMID- 14737972 TI - [Leukotrienes as inflammation mediators]. AB - Leukotrienes are biologically active metabolites derived from arachidonic acid playing an important role in inflammatory responses. There are two main groups of leukotrienes: dihydroxyleukotrienes (LTB4) and cysteinyl-leukotrienes (LTC4, LTD4, LTE4). By activating specific G-protein coupled receptors, leukotrienes take part in immune responses, like activation and chemotaxis of leukocytes. Several studies have shown that leukotrienes may play a significant role in pathomechanisms of inflammatory diseases of human airways, skin, digestive tract and heart. PMID- 14737974 TI - [Medical demography today and tomorrow]. PMID- 14737975 TI - [Demography of nephrologists in France in 2002]. AB - This survey was performed using data generated by a mailing sent with the collaboration of regional coordinators in 2002 to all the nephrologists identified in France. 1326 nephrologists were included in the pool, with an average of 22 nephrologists per million population (pmp), ranging from 14 to 29 pmp according to the different regions. Their mean (and median) age was 46.6 years, 30% were female. 63.5% of the nephrologists were working in a public hospital, 19.3% in private clinics, 13.3% in non-profit associations, 2% and 0.4% in research units or with industry, respectively. The data were used to generate a register of all the French nephrologists. 47 retirements per year are anticipated between 2010 and 2019, which yields an indication for the number of new nephrologists to be certified in this time-span. The gap between the future retirees and the newly trained nephrologists is very deep and cannot be bridged with the currently operating modes of recruitment. The increasing incidence and prevalence of patients with end stage renal failure will considerably increase the need of nephrologists, which is all the more amplified by the recent modification of the French law concerning the weekly upper limit of working time for physicians. Urgent measures have to be taken for preventing the consequences of dearth of nephrologists clearly anticipated for the next 10 years. PMID- 14737976 TI - [Pierre J. Desault and the birth of nephrology 1785-1795]. AB - Desault's genius and methods opened up a new field, the pathology of those medical conditions which afflict the urinary apparatus. Though he was a surgeon and therefore an anatomically based clinician, his most important discoveries were in pathological physiology, the polyuria of atrophic kidneys, the fatal dehydratation which follows the various forms of polyuric diabetes, and the oligo anurias which follow excess water loss from the gut, the lungs ... etc. In the last two conditions the kidneys might look normal and therefore might be presumed normal. If Desault's work had survived this physiological eruption into renal disorders would have been the first decisive step of modern nephrology. His message, though excellent, was delivered too soon and on barren soil, to poorly educated physicians who paid little attention. But then, one has to remember that Desault was just a surgeon! PMID- 14737977 TI - [Long term experience with the LifeSite hemodialysis device]. AB - We present the first report on the long term use of the LifeSite hemodialysis access system in patients who have no possibility of arteriovenous fistula creation. This system consist of a titanium alloy valve and silicone catheter which is placed in a central vein. Conventionally two devices are placed for optimal blood draw and return but single needle dialysis is possible using only one device. We have used this system in six patients since February 1999. Four patients continue to use the LifeSite for dialysis while one patient transferred to peritoneal dialysis because of hemodynamic instability during dialysis and one patient died of mesenteric infarction. Blood flow using two devices is excellent (300-345 ml/min). The infection rate is low (about 1.7 episodes per 1,000 days) and most patients were successfully treated without the need to remove the device. Two patients required temporary removal of one device because of local infection. While this, they were dialysed via a single needle using the remaining device. In our experience the LifeSite system appears a useful alternative in patients in whom a native fistula is not possible and it is able to provide effective dialysis for at least three years. PMID- 14737978 TI - [HLA-G: immunoregulatory molecule involved in allograft acceptance]. AB - The Human Leucocyte Antigen-G (HLA-G) is a non-classical MHC class I molecule of low polymorphism, restricted tissue distribution and tolerogeneic functions. It is clearly demonstrated that HLA-G contributes to fetal graft tolerance by the maternal immune system. The tolerogeneic properties of HLA-G act via specific inhibitory receptors present on immunocompetents cells: HLA-G inhibits natural killer cells (NK) and CD8+ T cell cytotoxicity, suppresses CD4+ T cell proliferation in response to allogeneic stimulation and promotes T helper 2 (Th2) type responses. The soluble HLA-G protein is spontaneously secreted by allo sensitized CD4+ T cells during mixed lymphocyte reactions (MLR), and inhibits their proliferative response. Finally, inhibition of dendritic cell maturation has been observed in HLA-G transgenic mice. In human organ transplantation, our group has reported in cardiac and liver-kidney transplanted patients, a positive correlation between the de novo ectopic expression of HLA-G in both patient's serum and graft biopsies, and a lower rate of acute rejection episodes of the grafts. Moreover no chronic graft rejection has been detected in those populations. These results support the involvement of HLA-G in regulatory mechanisms that may occur during human allotransplantation. PMID- 14737979 TI - [Cystatin C in the evaluation of renal function]. AB - Glomerular filtration rate (GFR) is the best indicator of renal function. GFR is usually estimated by serum creatinine or the creatinine clearance calculated on urine collected over 24 hours or with the Cockcroft formula. These methods are however limited. Serum creatinine has a very poor sensitivity and urine collection is difficult. Cystatin C is a protease inhibitor produced in a constant manner by nucleated cells. This molecule is freely filtrated by the glomerule and quite completely catabolized in the proximal tubules. Its plasmatic concentration might thus be used to estimate GFR. Presently available data allow to conclude that plasmatic cystatin C is at least as good as serum creatinine to estimate GFR. It is less sensible to changes in body mass. Its determination appears more sensitive to detect early mild changes in GFR. Reference values are presently available for the different methods of determination. Cystatin C plasma level determination is more expensive than routine creatinine plasma determination. In the absence of very significant advantages, this might explain its limited use in daily clinical practice. PMID- 14737980 TI - Renal disease in the acquired immunodeficiency syndrome in north central Nigeria. AB - BACKGROUND: The brunt of the human immunodeficiency virus infection/the acquired immunodeficiency syndrome is largely borne by communities in sub-Saharan Africa. We describe renal disease in Nigerians with the acquired immunodeficiency syndrome. METHODS: Consecutive patients with the acquired immunodeficiency syndrome (AIDS) seen in the infections unit of the Jos University Teaching Hospital and a similar group of healthy controls were evaluated for renal disease. Subjects with past history of renal disease, hypovolemia, hypertension, diabetes mellitus and/or a documented fever were excluded from the study. RESULTS: Of the 79 patients with the acquired immunodeficiency syndrome and 57 controls studied, renal disease was present in 41 (51.8%) of the patients in the AIDS group and 7 (12.2%) of controls. While 15 (19%) of the AIDS group had azotemia alone and 20 (25.3%) had proteinuria alone, 6 (7.6%) had azotemia and proteinuria. The mean protein excretion/24 hours was significantly higher in the AIDS group compared to controls, (2.99 +/- 54 g and 0.56 +/- 0.12 g respectively, p = 0.001), while the GFR was significantly higher in controls compared to the study group (103.30 +/- 37.78 and 68.03 +/- 37.55 respectively, p = 0.004). Subjects in the AIDS group with renal disease had a significantly longer duration of illness compared to those without (12.33 +/- 8.67 months and 7.28 +/- 7.78 months respectively, p = 0.008). Age and serum CD4+ cell counts were similar in patients with and without renal disease in the AIDS group. CONCLUSION: Renal disease is a common complication of acquired immunodeficiency syndrome, the duration of illness being strongly associated with its presence. PMID- 14737981 TI - Performance of normal Nigerian students on the mini-mental state examination. AB - BACKGROUND: Cognitive dysfunction is a feature of many diseases and the impact is especially relevant in the young. While there are many sophisticated tests of cognitive function, the shorter Mini Mental State examination (MMSE) is the most widely used screening test. In Nigeria where cognitive function is not routinely assessed in the management of cases, the MMSE is probably the most practical test of cognitive function to be promoted. This is because it is easy to administer and to interpret. There is however a need to define the performance of normal Nigerian subjects on the MMSE to make its use more relevant. METHODOLOGY: Seventy normal adolescents attending a secondary school in Ibadan, Nigeria, were administered the MMSE. Excluded were subjects with a history of mental impairment or medical disorders known to influence cognitive function. All subjects were tested only once. RESULTS: Normal MMSE scores were obtained by 88.6% of subjects while 11.4% of the subjects were found to be mildly or moderately cognitively impaired. More than 55% of the subjects did not obtain full scores with test of orientation in place while 30% and 34.3% respectively did not obtain full scores with tests of orientation in time and of attention respectively. Males and females performed equally with regard to the test items except for a non statistically significant better performance of the females with test of three item recall (p = 0.09). CONCLUSION: Abnormalities on the MMSE in Nigerians should be interpreted with caution as more than 10% of subjects may have abnormal scores. Tests of orientation and of attention are particularly likely to indicate abnormal cognitive function even in subjects with overall scores in the normal range. There may be a need to modify the MMSE to make it more applicable to young Nigerian subjects. PMID- 14737982 TI - Haematological profile and malarial parasitaemia in Nigerian children requiring emergency blood transfusion. AB - BACKGROUND: This study was carried out to determine the haematological profile and malarial parasitaemia of children requiring emergency blood transfusion. METHODS: This prospective study was carried out from 1st August to 30th of November, 1999 at the Children Emergency Ward (CHEW) of the Obafemi Awolowo University Teaching Hospital (OAUTH) Ile-Ife, Nigeria. All children requiring emergency blood transfusion seen at the CHEW were included in the study. Blood samples were taken on admission for haematocrit, white cell count and differentials, thin and thick blood films. Other data recorded include age, sex, clinical impression on admission. RESULTS: Four hundred children were admitted into the CHEW during the study period and 173(43%) required emergency blood transfusion. One hundred and forty-five (84%) of the children who required emergency transfusion were below the age of five years. There was no sex predilection. Although, clinical impression of malaria was made either singly or in combination with other diseases in almost all the patients, malaria parasitaemia was confirmed in 86 (50%) of the patients. The mean haematocrit was 0.14 L/L. In 60% of the children, there were mixed microcytic hypochromic and macrocytic red bloodcells. CONCLUSION: We conclude that severe anaemia requiring emergency blood transfusion is a major cause of morbidity and mortality in under five children. We also confirm that malaria and nutritional deficiencies are the major cause of severe anaemia in Nigerian children. We therefore recommend urgent need for prevention and prompt treatment of malaria in children under the age of five years. We also recommend the need for improvement in the nutritional intake of children under the age of five. PMID- 14737983 TI - Perceptions of occupational hazards amongst office workers at the Obafemi Awolowo University, Ile-Ife. AB - BACKGROUND: A Survey of 350 office workers at the Obafemi Awolowo University (OAU), Ile-Ife was conducted to determine respondents' perceptions of the occurrence of work-related hazards and to characterize the pattern of hazards found prevalent in the population. METHODS: The survey employed a descriptive cross-sectional design. Data were generated through the use of a structured self administered questionnaire and a non-participant observation checklist. RESULTS: Between 39% and 47% of the respondents expressed dissatisfaction with the level of control of the factors of the work environment (heat, noise, illumination, space, ergonomics). Two-thirds of respondents presented with work-related low backache, and this was more so for respondents who had spent at least 10 years on the job than those who had spent less. Of the 227 keyboard operators interviewed, 130 (57%) experienced symptoms suggestive of the overuse syndrome and 70% of the 82 computer operators interviewed had experienced varying degrees of visual fatigue since their appointments. CONCLUSION: The study concluded that the present design and layout of offices/workstations and access to equipment at the Obafemi Awolowo University, Ile-Ife is not optimal and tend to promote unnecessary physical efforts. This, in turn, might reduce efficiency and productivity. Suggestions for improvement were made as a way forward. PMID- 14737984 TI - Cervical vertigo and cervical spondylosis--a need for adequate evaluation. AB - BACKGROUND: Cervical spine spondylotic changes are known to cause vertigo. The mechanism by which cervical vertigo is induced is very debatable. This study was to highlight the prevalence of vertigo and other vestibulocochlear organ affectation in patients with cervical spondylosis and to serve as a template for further research in this area. METHODOLOGY: A six-year retrospective review of vertiginous patients with radiological evidence of cervical spondylosis seen at the Otorhinolaryngology Clinic of University College Hospital (UCH), Ibadan. RESULTS: Forty-three patients, 23 males (53.5%) and 20 females (46.5%) had vertigo as a symptom and radiological evaluation for cervical spondylosis. The radiological findings were normal in 11 patients (26%) and abnormal in 32 patients (74%). Of the twenty-five patients who had pure tone audiometry, 40% were normal and 60% abnormal. Out of 20 affected ears, seven patients (47%) had right, 3 patients (20%) left and 5 patients (33%) both ear involved respectively. High frequency range was affected in 8 (40%), low in 2 (10%) and all in 10 (50%) ears respectively. Decibel hearing level (dBHL) loss ranged from 30-90 dBHL with a mean binaural of 43 (+/- 8.5) dBHL. CONCLUSION: We advocate early multidisciplinary approach in the management of all established cases of cervical vertigo. PMID- 14737985 TI - Childhood morbidity and treatment pattern at the multipurpose health centre, Ilesa, Nigeria. AB - BACKGROUND: This is a descriptive case series study that attempt to identify the common presenting complaints why children are brought to the Multipurpose Primary Health Centre (MPHC) at Ilesa and also to examine the treatment pattern of the common illnesses from birth to the first five years of life, from October 2000 to March 2001. METHODS: Information was collected by reviewing the case records of all under five children who presented for treatment at the MPHC, Ilesa from October 2000 to March 2001 using a spreadsheet. The age, sex and presenting complaint as reported by their mothers were extracted from the records. The treatment given as recorded by the attending physician was also recorded. RESULTS: The findings of the study reveals that for every 20 outpatient visits to the centre, 11 were children under five years. The male to female ratio was 1:1. Overall, fever, cough and diarrhoea were the commonest presenting complaint (77.2% of all presenting complaints) with fever taking the highest percentage (48.8% of all presenting complaints) except for children under 6 months where majority present with cough (31.6% of the presenting complaint of that age group). The disease conditions most commonly treated by the physicians at the centre were malaria, acute respiratory tract infection and diarrhoea. The drugs commonly used are chloroquine, cotrimoxazole, and oral rehydration therapy. CONCLUSION: This study has shown that the major burdens of diseases in children under 5 years of age are preventable and are amenable to cost-effective and simple treatment. PMID- 14737986 TI - A monoblock resection for malignant phaeochromocytoma. AB - BACKGROUND: Phaeochromocytoma is a rare surgically treatable cause of hypertension. The aim of this paper is to present a case of phaeochromocytoma treated in Port Harcourt. METHOD: The case record of a patent with phaeochromocytoma and a review of the relevant literature. RESULT: A 40 year-old man presented with episodic malignant hypertension resistant to several anti hypertensive drugs. A 24-hour urinary Vanillyl Mandelic Acid estimation was high at 68 mmol. An ultrasound scan revealed a huge right suprarenal mass. Preoperative medication was given to reduce the blood pressure and prevent perioperative arrhythmias. Under general anaesthesia with propofol, the tumour was explored. It appeared to invade the kidney and there were multiple hepatic secondaries. It was resected in block with the kidney. Intra- and postoperatively he had episodes of hypertension which were successfully controlled with a combination of intravenous chlorpromazine 50 mg, tramadol 100 mg and lorazepam 4 mg. Histopathology examination showed that the suprarenal mass and hepatic lesions were identical showing malignant phaeochromocytoma. The post-operative period was satisfactory. Cytotoxic drugs were not given because they were not available. On review 8 weeks later, the patient remained well. CONCLUSION: Meticulous anaesthetic and surgical skills are essential in the resection of a phaeochromocytoma. PMID- 14737987 TI - Renal cell carcinoma presenting as a scalp swelling. AB - BACKGROUND: Renal cell carcinoma occasionally presents in an unusual manner. AIM: To report the rare presentation of renal cell carcinoma as a scalp swelling. METHOD: The case record of a patient with renal cell carcinoma and the literature on the disease were reviewed. RESULT: A 42-year old woman presented with a scalp swelling. Haematuria was missed at the first visit until she volunteered it on a subsequent visit. Investigations suggested renal cell carcinoma. This was confirmed following a right nephrectomy. The scalp lesion was a metastasis extending through the skull into the intracranial compartment. She was not given any systemic drugs because none was available. She survived until she was lost to follow up. CONCLUSION: Haematuria remains an important symptom of renal cell cancer and should be elicited during systemic enquiry. PMID- 14737988 TI - Drug errors in anaesthetic practice: case reports. AB - BACKGROUND: Error is inherent in any complex human endeavour. It is therefore not unusual for these to occur in drug administration. We report three cases of such errors. METHOD: The anaesthetic record of 3 patients who had errors in drug administration and discussion of the relevant literature. RESULT: Through the years in anaesthetic practice at the University of Port Harcourt Teaching Hospital (UPTH), our Pharmacy has always supplied standard ketamine hydrochloride in a dark brown bottle containing 10 mls of the 5% solution. Suxamethonium chloride on the other hand was always supplied in a 2 ml clear glass ampoule. A change in this known packaging without prior notification resulted in near misses, all of which had the potential for serious morbidity. CONCLUSION: A strict adherence to standard procedure is necessary to minimize the risk, since there is no substitute for vigilance if morbidity and mortality is to be prevented. PMID- 14737989 TI - From backward to forward clinical reasoning: time for a change. PMID- 14737990 TI - [Questionnaire-based survey on renal biopsy in Japan]. PMID- 14737991 TI - [Water metabolism and vasopressin]. PMID- 14737992 TI - [Salt intake and the progression of chronic renal diseases]. AB - To what extent dietary salt intake is involved in the pathogenesis of progressive renal diseases has never been fully understood in humans. To this aim, we investigated the relationship between urinary sodium excretion (under a low salt & low protein diet) and urinary protein excretion/renal function in patients with three major renal diseases: chronic glomerulonephritis(GN), diabetic nephropathy(DN) and nephrosclerosis(NS). The results were as follows; 1) A significant positive correlation was found between urinary sodium excretion (equivalent to the daily salt intake) and daily urinary protein excretion in patients with a GN and DN. However, no relationship was found between the two parameters in patients with NS. 2) Reduction in salt intake led to a significant decrease in daily protein excretion, the effect of which was prominent in patients with GN and DN. 3) A significant positive correlation was found between urinary sodium excretion and estimated protein intake(EPI) in all three groups. 4) There was a significant positive correlation between EPI and urinary protein excretion in DN, but not in GN. 5) Reduction in salt and protein intake(calculated as an EPI) ameliorates the slope of reciprocal creatinine concentration(1/Cr) in patients with GN and DN. These results indicate that slat restriction is strongly associated with the preservation of renal function in patients with GN and DN, suggesting that this dietary strategy can be a useful measure for retarding the progressive nature of these diseases. Of note is that both salt and protein restriction was renoprotective only in patients with DN. Thus, patients with GN and DN must be followed-up on the basis of a salt restricted diet throughout their clinical course. PMID- 14737993 TI - [Effect of oral intake of an enteric capsule preparation containing Bifidobacterium longum on the progression of chronic renal failure]. AB - Since the accumulation of intestinal putrefactive products, such as indole and phenol, is known to play a role in the exacerbation of chronic renal failure, reduction of these intestinal putrefactive products can be expected to retard the progression of renal failure. In the present study, an enteric capsule preparation containing Bifidobacterium longum(Bifidus HD) was administered orally to 27 patients with chronic renal failure(CRF) for 6 months. Though no significant effect was found in the whole group, a significant retardation of the progression of renal failure was found in patients with an initial serum creatinine level > or = 4.0 mg/dl or those with an initial serum inorganic phosphate level > or = 4.0 mg/dl. There was no adverse effect observed in any case. Bifidus HD is considered a useful tool for suppressing the progression of chronic renal failure(CRF) in the conservative period. PMID- 14737994 TI - [Two cases of lupus cystitis complicated by lupus nephritis treated successfully with steroid therapy]. AB - In patients with systemic lupus erythematosus(SLE), interstitial cystitis(lupus cystitis) is an uncommon, but important manifestation. We report two Japanese patients with lupus cystitis. Case 1 was a 49-year-old woman diagnosed as having rheumatoid arthritis and membranous nephropathy. She was treated with prednisolone(5 mg daily). Case 2 was a 41-year-old woman also diagnosed as having rheumatoid arthritis previously and treated with a non-steroidal anti inflammatory drug. Both cases presented abdominal pain, vomiting, dysuria and frequency of micturition. We diagnosed these cases as SLE on the basis of arthritis, renal disorder(proteinuria and hematuria), and positive antinuclear and anti-dsDNA antibodies. In addition, bilateral hydronephrosis was found in both cases. Thus, they were also diagnosed as probable lupus cystitis. The patients were treated with one cycle of methylprednisolone pulse therapy. Thereafter they were treated with 60 mg/day of prednisolone and their symptoms resolved promptly. Furthermore, no abnormal finding was found by abdominal ultrasonography and/or the intravenous pyelogram after therapy. Renal biopsies were performed and both cases showed lupus glomerulopathy (case 1: WHO class Vb, case II: WHO class IVb). Abdominal pain and/or dysuria, which is common in SLE patients, requires further examinations to evaluate the lupus cystitis. PMID- 14737995 TI - [A case of membranous nephropathy with ANCA-associated necrotizing glomerulonephritis during oral administration of PTU for Graves' disease]. AB - We experienced a coincidental case of two types of glomerulopathy associated with Graves' disease. A 64-year-old man, who had been treated with propylthiouracil(PTU) for Graves' disease for 15 years, was admitted to our hospital for macroscopic hematuria and rapidly progressive deterioration of renal function. Although his thyroid function had been within the normal range during treatment, the level of thyrotropin receptor antibody(TRAb) gradually increased from a year before admission. Serological tests revealed that he was positive for myeloperoxidase-antineutrophil cytoplasmic antibody(MPO-ANCA). The renal biopsy specimen showed necrotizing and crescentic glomerulonephritis(GN) superimposed on membranous nephropathy(MN). This is a rare case of MN complicated with ANCA associated crescentic GN in a Graves' disease patient. Association of these two renal alterations was not clearly defined. MN involved with Graves' disease also has been rarely reported. Some reports demonstrated deposition of thyroglobulin and other thyroid related antigens in the glomeruli. In the present case, long term impairment of Graves' disease and elevation of TRAb might have been responsible for the formation and deposition of thyroid-associated immune complex in the glomeruli. As for crescentic GN, PTU might have induced ANCA-associated GN independently of MN. This case is instructive for considering the relation between Graves' disease and renal injury. PMID- 14737996 TI - Medication management in low care residential aged care facilities. PMID- 14737997 TI - Australians want health care not welfare. PMID- 14737999 TI - [Transfer of medical competences, an opportunity for the profession?]. PMID- 14738000 TI - [An opening for paramedics. Interview by Celine Dekussche]. PMID- 14738007 TI - [I--Phases of scarring 3/3 Granulation tissue]. PMID- 14738008 TI - [Note on wounds and dressings. 5--Hydrogels]. PMID- 14738009 TI - [Integrating relationship management with clinical management]. PMID- 14738010 TI - [In rehabilitation, consider the patient more than the illness]. PMID- 14738011 TI - [Relationship care or relation of care?]. PMID- 14738012 TI - [Hospitalization, a space for possibilities]. PMID- 14738013 TI - [Communicating without words, using the five senses]. PMID- 14738014 TI - [Using the body to communicate]. PMID- 14738015 TI - [Psychological care in intensive care]. PMID- 14738017 TI - [Recognize the fragility of the patient]. PMID- 14738016 TI - [The erl king or the therapeutic relationship]. PMID- 14738018 TI - [All the colors of a smile]. PMID- 14738019 TI - [9/10 Coffee]. PMID- 14738020 TI - [II--Relationship care]. PMID- 14738021 TI - Types of hysterectomy. Comparison of characteristics, hospital costs, utilization and outcomes. AB - OBJECTIVE: To compare hospital costs, patient characteristics and outcomes of 3 hysterectomy techniques--abdominal, vaginal and laparoscopically assisted vaginal (LAVH). STUDY DESIGN: A cross-sectional analysis was performed using patients discharged from Florida hospitals in 2000 with hysterectomy as the primary procedure. To avoid differences due to unrelated complications, records indicating cancer or other major non-hysterectomy-related procedure were excluded from the analysis. A total of 23,191 records were used to compare the 3 techniques on hospital costs and length of stay, controlling for patient differences in complicating diagnoses and related procedures. RESULTS: Consistent with previous studies, patients undergoing LAVH had higher hospital costs, shorter lengths of stay and no difference in surgical complications from either vaginal or total abdominal hysterectomy, even after controlling for patient comorbidities. Other factors affecting hospital costs and length of stay were patient race, type of insurance, hospital ownership and location. CONCLUSION: LAVH is still more expensive than vaginal and total abdominal hysterectomy but offers a speedier recovery, with no measurable difference in the rate of complications. Further research is warranted to ascertain differences in readmission rates across the techniques and to investigate the process used to select which hysterectomy technique is used for a given patient. PMID- 14738022 TI - Bilateral ascending uterine artery ligation vs. tourniquet use for hemostasis in cesarean myomectomy. A comparison. AB - OBJECTIVE: To comparatively investigate the effects of using bilateral ascending uterine artery ligation and tourniquet use on intraoperative and postoperative blood loss during myomectomy in cesarean cases. STUDY DESIGN: A total of 70 pregnant women diagnosed with myomas in the prenatal period were included in this randomized, prospective study. They were admitted to our department during the study period. Fifty-two patients who underwent cesarean myomectomy were randomly divided into 2 equal groups. In the first group bilateral ascending uterine artery ligation and myomectomy were performed after lower uterine segment transverse cesarean section. The second group served as the control group; myomectomy was performed with a tourniquet. For statistical analysis, Mann Whitney U, chi 2 and Wilcoxon Rank tests were used. Spearman correlation analysis (rs, n, p) was used for analysis of correlation between the duration of the myomectomy operation and blood loss and number of enucleated myoma nuclei during myomectomy. RESULTS: Total intraoperative blood loss, total operation duration, number of enucleated myoma nuclei (Mann Whitney U test) and febrile morbidity (chi 2 test) were similar in the 2 groups (P > .05). A significant positive correlation was established between the duration of the myomectomy operation and loss of blood and number of enucleated myoma nuclei during myomectomy (rs = .9, n = 52, P = .000). Urgent laparotomy and bilateral internal iliac artery ligation had to be performed in 1 patient in the tourniquet group who had a postoperative hemorrhage. CONCLUSION: Despite the fact that bilateral ascending uterine artery ligation and tourniquet use had similar outcomes with regard to intraoperative blood loss in cesarean myomectomy cases, the efficacy of ligation on blood loss in the postoperative period continues owing to its permanence. The tourniquet method is not effective in the postoperative period since the tourniquet is removed at the end of the operation. Therefore, bilateral ascending uterine artery ligation may be preferable in cesarean myomectomy cases. PMID- 14738023 TI - Familial aggregation of type 2 diabetes and chronic hypertension in women with gestational diabetes mellitus. AB - OBJECTIVE: To perform a case-control study to assess the extent to which women with a positive parental history of type 2 diabetes and/or chronic hypertension experienced an increased risk of developing gestational diabetes mellitus (GDM). STUDY DESIGN: Participants (133 GDM cases and 373 controls) provided information on first-degree family history of the 2 conditions and other covariates of interest in interviews. Logistic regression procedures were used to derive odds ratios and 95% confidence intervals adjusted for confounding by maternal age, race/ethnicity and prepregnancy adiposity. RESULTS: As compared with women with no parental history of diabetes, women with a maternal-only (odds ratio = 2.0), paternal-only (odds ratio = 2.3) or both maternal and paternal history of diabetes (odds ratio = 3.8) experienced a statistically significant increased risk of GDM. The odds ratio for women with a positive parental history of diabetes and hypertension was 2.6 (95% confidence interval, 1.3-5.3). Women with a diabetic sibling had an 8.4-fold increased risk of GDM (95% confidence interval, 2.1-33.4). First-degree family history of chronic hypertension was predictive of GDM risk but only when hypertension was associated with a diagnosis of diabetes. CONCLUSION: Our results are consistent with the thesis that family history of diabetes (alone or when associated with hypertension) reflects genetic and behavioral factors whereby women may be predisposed to an increased GDM risk. PMID- 14738024 TI - Relationship of paternal factors to birth weight. AB - OBJECTIVE: To investigate the relationship between paternal characteristics and birth weight. STUDY DESIGN: A total of 241 gravidas with uncomplicated, singleton, term pregnancies were studied. Maternal demographic and pregnancy specific characteristics were used to calculate the expected birth weight for each fetus using a previously validated birth weight prediction equation. The additional independent predictive value of 4 paternal variables was assessed using multiple regression. RESULTS: Before adjustment for other variables, paternal height and weight significantly correlated with birth weight, but paternal age and body mass index did not. After controlling for maternal and pregnancy-specific factors that are known to influence fetal weight, only paternal height was significant as a predictive variable. The proportion of variance in birth weight that could be independently explained by paternal height was 2%. A 10-g gain in fetal weight was associated with each centimeter of increase in paternal height (P < .02). Using the resulting combination equation that included paternal height as a variable, 31% of the variance in term birth weight could be explained, and birth weights could be accurately predicted to within +/- 8.3% (+/- 288 g). Fathers with heights 2 SD above and below the mean had the term birth weight of their offspring increased and diminished by 125 g, respectively. CONCLUSION: Paternal height explains an independent portion of the variance in term birth weight among normal newborns of up to 250 g that cannot be explained by other maternal or pregnancy-specific factors. Paternal age, weight and body mass index do not independently influence birth weight. PMID- 14738025 TI - Maternal views and acceptance of antenatal HIV screening in a university teaching hospital in Hong Kong. AB - OBJECTIVE: To assess maternal acceptance, knowledge, attitude, perceived risks and barriers toward antenatal HIV screening. STUDY DESIGN: Prospective anonymous survey of 1,519 pregnant women recruited in a university teaching hospital RESULTS: Women demonstrated fairly good knowledge of and a positive attitude toward HIV screening. Condom usage was 61.4%, and 25.3% of women had at least 1 risk factor for HIV infection. Support for mandatory and universal screening was 31.4% and 48.8%, respectively. A total of 82.6% women agreed to HIV testing. The major reason for declining the test was that women considered themselves to be at low risk (84.3%). Women with risk factors tended to prefer more aggressive methods of antenatal testing (P < .001) and more readily accepted HIV screening (89.8% vs. 73.1%, P < .001). CONCLUSION: Given the high acceptance rate in our local population, universal offering but voluntary testing is the optimal mode of antenatal HIV screening in Hong Kong. PMID- 14738026 TI - Emergency cesarean delivery for nonreassuring fetal heart rate tracings. Compliance with ACOG guidelines. AB - OBJECTIVE: To review the English-language literature from 1990 to 2000 on cesarean delivery for "fetal distress" and assess compliance with American College of Obstetricians and Gynecologists (ACOG) guidelines. STUDY DESIGN: A PubMed search with the search items cesarean, fetal distress, cesarean, nonreassuring fetal heart rate, cesarean, neonatal acidosis and cesarean, umbilical arterial pH was undertaken. Excluded from the search were case reports, letters to the editor, focus on fetal anomaly, combinations with other reasons for operative delivery (either abdominally or vaginally) or absence of pertinent information. RESULTS: Among 392 articles from the search, 169 met the inclusion criteria. Three reports provided detailed information on use of scalp pH; use occurred in 5% (60/1,128) of emergency cesareans. Three reports provided data on the use of tocolytics for intrauterine resuscitation; the combined result was 16% (201/1,261). Five reports assessed the decision-to-incision interval within 30 minutes; in 59% (262/446) of cases it was achieved. Five reports contained cord pH information on 340 emergency cesarean deliveries; umbilical arterial pH was < 7.00 in 10%. CONCLUSION: Physician use of, and compliance with, ACOG guidelines for emergency cesarean deliveries is difficult to assess, and incomplete compliance appears commonplace. PMID- 14738027 TI - Clinical value of a single serum CA-125 level in women with symptoms of imminent abortion during the first trimester of pregnancy. AB - OBJECTIVE: To assess the clinical value of a single serum CA-125 level in women presenting with symptoms of imminent abortion from 5 to 12 weeks' gestation. STUDY DESIGN: Two hundred women with symptoms of imminent abortion from 5 to 12 weeks' gestation were allocated to 3 subgroups: group I (n = 50), abdominal pain only; group II (n = 80), abdominal pain and vaginal bleeding of less than 3 days' duration; and group III (n = 70), abdominal pain and vaginal bleeding of > or = 3 days' duration. The control group (n = 55) was women undergoing a normal pregnancy of comparable gestational age. In all patients, CA-125 and beta-human chorionic gonadotropin (beta-hCG) levels were determined with commercially available enzymatic immunoassay tests. RESULTS: A significant increase in serum CA-125 levels was observed in patients with vaginal bleeding (groups II and III) in comparison with the control group. In contrast to CA-125, serum beta-hCG levels were significantly lower in patients who went on to miscarry. Receiver operating characteristic curve analysis revealed that all women with symptoms of imminent abortion who have a CA-125 level of > or = 43.1 U/mL should be considered as having a greater risk of miscarriage. Patients with bleeding of > or = 3 days should have a threshold value of 66.5 U/mL applied as a risk determinant. CONCLUSION: A single serum CA-125 level determination is valuable in women with imminent abortion presenting with abdominal pain, vaginal bleeding or both. PMID- 14738028 TI - Conservative management of vesicouterine fistula. A report of 2 cases. AB - BACKGROUND: Vesicouterine fistulas usually require laparotomy for repair. CASES: A vesicouterine fistula occurring after cesarean section was successfully managed hormonally. In another case it developed in association with an intrauterine device and was repaired translaparoscopically. CONCLUSION: Laparotomy may be avoidable in the management of a vesicouterine fistula. PMID- 14738029 TI - Uterine incarceration in a 9-week multifetal pregnancy resulting from in vitro fertilization. A case report. AB - BACKGROUND: Although retroversion of the uterus is a common finding, incarceration of the gravid uterus is a rare complication. It occurs usually between the 12th and 16th weeks of gestation and can lead to severe complications. No case of incarceration has been reported before in a multifetal gestation resulting from in vitro fertilization. CASE: A 35-year-old woman with a triplet pregnancy presented at 9 weeks' gestation with acute urinary retention and was diagnosed with uterine incarceration. This diagnosis was confirmed by ultrasound and was resolved using transvaginal pressure under epidural anesthesia. CONCLUSION: This is the first reported case of uterine incarceration in a triplet pregnancy that resulted from in vitro fertilization. There is no universal protocol for treatment of incarceration, but several techniques have been described. PMID- 14738030 TI - Association between cervical schistosomiasis and cervical cancer. A report of 2 cases. AB - BACKGROUND: HPV is now considered the most important risk factor for the development of cervical intraepithelial neoplasia (CIN) and cancer. Although CIN and cancer have been previously reported in association with cervical schistosomiasis, those reports failed to control for the potential coexistence of high-risk HPV. CASES: Two women, 1 with high grade CIN and 1 with invasive cervical cancer, were negative for high-risk HPV subtypes. Evidence of cervical and systemic schistosomal infestation was evident in both cases. CONCLUSION: In support of prior published studies, cervical schistosomiasis seems to be a possible risk factor for the development of CIN and cancer. As populations around the world migrate, North American colposcopists need to become aware of this association. PMID- 14738032 TI - Key issues in transforming health care organizations for quality: the case of advanced access. AB - BACKGROUND: The 2001 Institute of Medicine (IOM) report highlighted the need for transformation of the U.S. health care system. This rigorous qualitative evaluation of transformational change for patient access in one large multispecialty group practice identifies the major issues facing organizations addressing the IOM challenge. METHODS: Semistructured depth interviews were conducted with the medical and administrative leaders at all levels, physicians, and nurses from 17 primary care clinics in one integrated medical group two years after they began to transform their approach to primary care patient appointment access. RESULTS: The mean time to third-next-available appointment was reduced by 76% during one year, from 17.8 days to 4.2 days. Nine important issues related to the change process were identified from clinic interviews. When combined with issues identified by central leaders, 13 themes stood out as lessons in transformational change. A major issue is the tension between physician autonomy and both effective organizational function and putting patients first. Physician autonomy is also diminished by the need to standardize and systematize care. CONCLUSIONS: Transformational change in care delivery is possible in large and complex group practices. Changes that directly affect care delivery and physician autonomy present particular challenges to physicians that need to be attended to if the changes are to be successful. PMID- 14738031 TI - Standardization as a mechanism to improve safety in health care. AB - BACKGROUND: A growing body of literature shows that when patterns of care are widely divergent, clinical outcomes suffer and, as a result, safety may be compromised. A multispecialty group at Luther Midelfort, Mayo Health System (LM, MHS) initiated efforts to reduce variance in the clinical practice patterns of providers. The pilot initiative, which entailed standardization of a sliding scale insulin protocol, served as a template throughout the LM, MHS for reducing variance and enhancing safety. STANDARDIZING INSULIN ADMINISTRATION: A single sliding-scale insulin protocol for regular insulin use in appropriate patients was intended to decrease the number of hypoglycemic events. A six-week comparison revealed that in the protocol-driven standardized sliding-scale insulin group, two episodes of hypoglycemia occurred in 134 dosages administered versus 20 hypoglycemic events in 519 dosages administered in the traditional group (1.49 versus 3.85%, p < .04). Subsequent 30-month data months revealed a reduction in hypoglycemic episodes from 2.95% to 1.1%. MEDICATION USE PROBLEM: A reconciliation of medications initiative focused on clarifying, correcting, and specifying the medications patients were consuming at different intervals of their hospitalization and then amending the data in the medical record. In a seven-month chart audit, errors per 100 admissions decreased from 213 to fewer than 50 errors. DISCUSSION: Standardization efforts to increase uniformity of practice are worth considering in other practice areas to increase safety and possibly reduce costs. PMID- 14738033 TI - Which strategies facilitate improvement in quality of care for elderly hospitalized pneumonia patients? AB - BACKGROUND: A retrospective cohort study was conducted to elucidate which hospital-based quality improvement (QI) strategies are most effective in facilitating improvement in care for patients with community-acquired pneumonia. METHODS: In 1999 telephone interviews were conducted with 29 acute care hospitals in Connecticut regarding their use of QI strategies for 1,234 patients at baseline and 1,081 patients at follow-up. RESULTS: Hospital-based QI strategies were grouped into two domains of implementation approach (epidemiologic and social influence). Hospitals scoring a 4 in either the epidemiologic or social influence approach (versus a score of < or = 3) experienced a greater-than average increase in percentage of patients with blood culture collection within 24 hours of hospital presentation. Hospitals applying all four social influence QI strategies showed a greater-than-average increase in delivery of antibiotics within 8 hours of patients' hospital arrival when compared with all the other hospitals combined. DISCUSSION: The finding that an increased proportion of patients receiving antibiotics within 8 hours and blood cultures within 24 hours of hospital arrival when the greatest numbers of hospital-based QI strategies were implemented in suggestive of a possible "dose effect" of QI. PMID- 14738034 TI - Improving ambulatory surgical pain management. AB - A surgical pain improvement team used an improvement model to standardize pain management at two hospital-based ambulatory surgery facilities. PMID- 14738035 TI - Using patient waiting-time data to improve the hospital bed-assignment process. AB - An improvement activity involving use of real-time waiting-time data resulted in reductions in bed-assignment times and overall diversion hours. PMID- 14738036 TI - The roles of government in improving health care quality and safety. AB - BACKGROUND: Discussions surrounding the role of government have been and continue to be a favorite American pastime. A framework is provided for understanding the 10 roles that government plays in improving health care quality and safety in the United States. Examples of proposed federal actions to reduce medical errors and enhance patient safety are provided to illustrate the 10 roles: (1) purchase health care, (2) provide health care, (3) ensure access to quality care for vulnerable populations, (4) regulate health care markets, (5) support acquisition of new knowledge, (6) develop and evaluate health technologies and practices, (7) monitor health care quality, (8) inform health care decision makers, (9) develop the health care workforce, and (10) convene stakeholders from across the health care system. CONCLUSION: Government's responsibility to protect and advance the interests of society includes the delivery of high-quality health care. Because the market alone cannot ensure all Americans access to quality health care, the government must preserve the interests of its citizens by supplementing the market where there are gaps and regulating the market where there is inefficiency or unfairness. The ultimate goal of achieving high quality of care will require strong partnerships among federal, state, and local governments and the private sector. Translating general principles regarding the appropriate role of government into specific actions within a rapidly changing, decentralized delivery system will require the combined efforts of the public and private sectors. PMID- 14738037 TI - Explaining low ratings of patient satisfaction among Asian-Americans. AB - Asian-Americans report lower levels of satisfaction with health care than other racial or ethnic groups. To determine potential reasons for this observation, we analyzed data from a national household telephone survey examining various aspects of health and health care across racial or ethnic groups. As in previous studies, Asians reported lower satisfaction with their physicians and health care services than other racial or ethnic groups, even after adjusting for demographic and health-related variables. However, Asians also reported lower satisfaction with their health insurance and with life in general and were less likely than non-Asians to have changed physicians due to dissatisfaction. We conclude that lower satisfaction ratings among Asians may be due to different response tendencies rather than to differences in quality of care. Further research is needed to explain this phenomenon and assess its potential impact on quality-of care evaluations for clinicians and health care organizations providing care for large numbers of Asian-Americans. PMID- 14738038 TI - By the numbers. Et cetera. PMID- 14738039 TI - [Hexavalent vaccines and fatalities. Causal relationship?]. PMID- 14738040 TI - [Therapy of hypertension. Lasting organ protection moves into the foreground]. PMID- 14738041 TI - [Call for attention to hemophagocytic syndrome]. PMID- 14738042 TI - [We need more general internists in China]. PMID- 14738043 TI - [Community hospital should set up general internal medical unit]. PMID- 14738044 TI - [The use of glucocorticosteroids in the treatment of severe acute respiratory syndrome]. PMID- 14738045 TI - [Be vigilant of the adverse reactions of glucocorticosteroids therapy]. PMID- 14738046 TI - [Some opinions on the use of glucocorticosteroids in severe acute respiratory syndrome]. PMID- 14738047 TI - [Personal views on treating severe acute respiratory syndrome with glucocorticosteroids]. PMID- 14738048 TI - [Coexistence of normal pregnancy and gestational trophoblastic neoplasia]. PMID- 14738049 TI - [Current classification an appellation of pregnancy induced hypertension]. PMID- 14738050 TI - [Genome research on susceptibility loci in inflammatory bowel disease]. PMID- 14738051 TI - [Radioactive synoviorthesis for the treatment of chronic haemophilic synovitis]. PMID- 14738052 TI - [What can we learn from irritable bowel syndrome?]. PMID- 14738054 TI - [Open up a new prospect on research and clinical application of mechanical ventilation]. PMID- 14738053 TI - [Controversial effects of aspirin on coronary artery disease and chronic heart failure]. PMID- 14738055 TI - [The need to enhance general internal medicine]. PMID- 14738056 TI - [Introduction of related indices in the evaluation of periodicals]. PMID- 14738061 TI - Managing sickle-cell crisis. PMID- 14738062 TI - Drawing blood with vacuum tubes. PMID- 14738066 TI - What is osteonecrosis? PMID- 14738067 TI - Administering smallpox vaccine: a two-pronged risk. PMID- 14738068 TI - Myths and facts...about bipolar disorder. PMID- 14738069 TI - [The language of cook books of the late middle ages and early modern times]. PMID- 14738070 TI - Elements of the medical emergency's epistemological alignment: 18-20th-century perspectives. AB - This paper develops the concept of epistemological alignment as a means of understanding how the medical emergency entered into biomedical classification. The working hypothesis is that the medical emergency could not develop as a paradigm from which knowledge could proliferate until a series of related concepts has been elucidates, and its epistemology became aligned. This argument is advanced by exploring the historically constructed lexicon of the medical emergency's key components. In the process, a semantic network created by 18th century attempts to problematize sudden death is elucidates and explored. The paper concludes by arguing that the First World War served as a catalyst for the alignment of concepts central to the epistemology of the medical emergency. PMID- 14738071 TI - Professionalization, gender and female-dominated professions: dental hygiene in Ontario. AB - This paper explores the influence of gender and feminism on the professional projects of female-dominated professions, through a case study of dental hygiene in Ontario. Full professional status has eluded many female-dominated professions, including dental hygiene. Historically, dental hygiene was defined as work for women, to be performed strictly under the control of male dentists. In recent years, dental hygiene has pursued a professional project and struggled for greater independence from dentistry. Ideas about gender and, particularly, feminism have been central to their professional project. PMID- 14738072 TI - "Would you let your child die rather than experiment on animals?" A comparative questions approach. AB - By placing the title question alongside five comparative questions and offering answers to the whole set as given by seven imaginary respondents, this paper analyzes the question's deceptiveness and the inconsistency of its implied claims. Apart from ambiguities of situation, history, and agency, the question's demand for a choice between "your child" and "nonhuman animals" obscures a field of other values regarding (1) species, (2) family ties, and (3) the wrongness-in itself (or otherwise) of the experiments envisioned. This paper argues that while a "No" answer to the title question does not, as intended by the questioner, support the experimental status quo, even a "Yes" answer does not reflect a choice between one's own child and animals. PMID- 14738073 TI - Ann Taylor's "The maniac's song": an unacknowledged source for Keats's "La belle dame sans merci". PMID- 14738074 TI - Clinical features suggestive of lymphadenopathy in a painting by Marinus Van Reymerswaele. PMID- 14738075 TI - [The medical plants and the description of the "enfermedad de montana" in the Natural and moral history of India (1590) of Jose de Acosta]. PMID- 14738076 TI - Identifying secondary skin lesions. PMID- 14738078 TI - The force of ideas. PMID- 14738079 TI - Are research schools necessary? Contrasting models of 20th century research at Yale led by Ross Granville Harrison, Grace E. Pickford and G. Evelyn Hutchinson. AB - This paper compares and contrasts three groups that conducted biological research at Yale University during overlapping periods between 1910 and 1970. Yale University proved important as a site for this research. The leaders of these groups were Ross Granville Harrison, Grace E. Pickford, and G. Evelyn Hutchinson, and their members included both graduate students and more experienced scientists. All produced innovative research, including the opening of new subfields in embryology, endocrinology and ecology respectively, over a long period of time. Harrison's is shown to have been a classic research school; Pickford's and Hutchinson's were not. Pickford's group was successful in spite of her lack of department or institutional position or power. Hutchinson and his graduate and post-graduate students were extremely productive but in diverse areas of ecology. His group did not have one focused area of research or use one set of research tools. The paper concludes that new models for research groups are needed, especially for those, like Hutchinson's, that included much field research. PMID- 14738080 TI - Organ weight effects of drowning and asphyxiation on the lungs, liver, brain, heart, kidneys, and spleen. AB - An examination of the organ weights associated with victims of drowning, asphyxiation and trauma was undertaken to determine (a) the effects of asphyxiation compared to a trauma group, and in turn, (b) the effects of drowning compared to an asphyxiation group. Included in the study were 217 drowning deaths, 166 pure asphyxiation deaths and 381 trauma deaths. The effects of asphyxiation (compared to trauma) resulted in elevated mean organ weights for the lungs, liver, kidneys and spleen (with mean increases of 17.8, 10.5, 10.3 and 23.4%, respectively). Effects of drowning (compared to asphyxiation) resulted in elevated mean organ weights only with the lungs and kidneys (with mean increases of 30.0 and 4.4%, respectively). Only the mean heart and brain weight remained constant across all experimental groups. A picture of drowning is suggested in which elevated lung and kidney weights are the result of both asphyxiation and the aspiration of water that occurs with drowning, whereas elevated spleen and liver weights in drowning victims are associated with only the effects of asphyxiation. In addition, the common autopsy finding of a small, anemic spleen in drowning, rather than caused by some pathophysiological mechanism of death, is hypothesized to be a postmortem phenomenon. PMID- 14738081 TI - Percutaneous drainage and explanation of an infected aortic endoluminal stent graft. AB - Endoluminal repair of abdominal aortic aneurysms (AAAs) has undergone explosive growth in the last decade. Although immediate benefits are attractive to both the patient and the treating physician, concerns regarding long-term success have dampened sustained enthusiasm for this technique. A rare but catastrophic complication is stent graft infection. This case report describes an early infection of an endoluminal aortic stent graft treated initially with percutaneous drainage and ultimately with staged extraanatomic bypass and graft explantation. PMID- 14738082 TI - Science as ideology: the rejection and reception of sociobiology in China. AB - The spread of sociobiology in China is not simply an internal event in the development of science. From the day it was introduced to China, its destiny was closely bound up with the development and change of Chinese society. Although it did not create as great disturbance as in America, it did have a significant impact in academic circles. However, scholars have paid little attention to these historical events. Today, sociobiology seems outdated and Wilson's grand agenda seems to have faded with the passing of time, but the mark that he made on the history of science is still striking. A review of the process of reception of sociobiology in China is helpful for understanding this theory as well as the society of China. PMID- 14738083 TI - Multiple mycotic aortic aneurysms due to Streptococcus pneumoniae. AB - Primary pneumococcal aortic mycotic aneurysms are rare clinical entities. Only a few cases have been reported in the literature. Extremely rare presentation is the occurrence of three simultaneous aneurysms. Treatment usually necessitates intravenous antibiotherapy combined with staged surgical interventions. This report highlights the case of a 52-year-old man with multiple Streptococcus pneumoniae mycotic aneurysms that were simultaneously and successfully treated during a one-stage surgical procedure. The aorta may be prone to infection, especially when its intima is structurally altered by pathologic processes like atherosclerosis, inflammation or trauma. Mycotic aneurysm is a rare but serious vascular condition needing urgent medical and surgical attention because of potential lethal complications. PMID- 14738084 TI - Measuring adherence to antihypertensive drug therapy. PMID- 14738085 TI - Endovascular exclusion of a postendarterectomy carotid pseudoaneurysm. AB - Carotid pseudoaneurysms are rare occurrences. They often result from trauma, but can also present following carotid endarterectomy. Treating such pseudoaneurysms can be difficult due to previous surgery and limited access to the high internal carotid artery. A case involving a postendarterectomy carotid pseudoaneurysm treated via a femoral approach with a covered stent using endoluminal techniques is presented. PMID- 14738086 TI - A true aneurysm of axillary-subclavian artery with cystic medionecrosis: an unusual manifestation of Marfan syndrome. AB - We report a case of a 69-year-old female patient diagnosed with an axillary subclavian artery(ASA) aneurysm, 7 cm long and 4 cm in diameter. The aneurysm had recently developed during follow-up for aortic sinus dilation associated with Marfan syndrome, which had been diagnosed in 1987. The patient underwent corrective surgery for the ASA aneurysm, and the aneurysm was histologically diagnosed as a true type with cystic medionecrosis. PMID- 14738087 TI - Leiomyosarcoma of the major peripheral arteries: case report and review of the literature. AB - We describe a case of primary intramural, extraluminal leiomyosarcoma of the common iliac artery. The patient presented with pelvic and leg pain and pheochromocytoma-like symptoms. The diagnosis was made intraoperatively, the tumor was resected, vascular reconstruction was performed, and postoperative radiation therapy was administered. Primary leiomyosarcoma of a major peripheral artery is extremely rare, and this report examines the clinical presentation, treatment, and prognosis of these patients. PMID- 14738088 TI - Arteriovenous PTFE dialysis access in the lower extremity: a new approach. AB - Advances in hemodialysis technology have allowed application of this treatment modality to an ever-increasing number of patients. Maintaining patients on dialysis for more than a decade is not unusual and the treatment of such patients may be limited primarily by the availability of vascular access. The aim of this study was to assess the efficacy and safety of a PTFE bridge graft between the superficial femoral artery and vein when more conventional arteriovenous fistulas cannot be accomplished. Use of the lower extremity had previously been avoided because of a greatly increased risk of infection. Over the period October 1, 1992 to March 31,1997, we performed 14 of these grafts in 11 patients (6 men and 5 women) whose median age was 56 (range 34-85) years. The patients were on long term hemodialysis and vascular access in the upper extremity was not possible. The median time from start of hemodialysis to receiving a suprapatellary bridge graft was 8 months (range 0-67). The suprapatellary graft was performed on two patients who were just starting on hemodialysis. Six patients received PTFE prosthetic implants in both arms and the other three patients underwent both native arteriovenous fistulas and PTFE prosthetic implants in the arm before the thigh was used. Only one patient had diabetes mellitus. Primary patency at 1 year was 54% and at 2 years, 18%. Secondary patency at 1 year was 64% and at 2 years, 18%. A PTFE bridge graft between the superficial femoral artery and vein for hemodialysis is a promising alternative when upper extremity arteriovenous fistulas cannot be constructed. The procedure is easily and rapidly performed with good access, and the patient clientele is easily selected. The technique has not been described previously. PMID- 14738089 TI - Stenosis at the internal carotid artery origin. PMID- 14738090 TI - Complex questions embedded in tracheotomy decisions. PMID- 14738091 TI - Conservative management of MRSA periaortic graft abscess. AB - Following repair of ruptured abdominal aortic aneurysm a patient developed methicillin-resistant Staphylococcus aureus perigraft abscess. The patient's clinical condition did not permit conventional open surgical correction. Ultrasound-guided aspiration and insertion of percutaneous drain enabled local antibiotic irrigation accompanied by systemic therapy. At 2 years follow-up the patient remains well with no evidence of sepsis. PMID- 14738092 TI - Endovascular management of a peroneal artery injury due to a military fragment wound. AB - A healthy 29-year-old male suffered bilateral lower extremity fragment wounds resulting in a tense right calf shortly following a wound that required a four compartment fasciotomy. During treatment of other injuries after medical evacuation he complained of increasing right calf pain. A peroneal pseudoaneurysm (PSA) with a concomitant arteriovenous fistula (AVF) was diagnosed. The distal one-third of the peroneal artery was not visualized, and the posterior tibial and anterior tibial arteries were normal. The patient underwent percutaneous coil embolization of the pseudoaneurysm. Completion arteriography demonstrated no evidence of the PSA and AVF. The patient's bruit and pain resolved. PMID- 14738093 TI - Rupture of abdominal aortic aneurysms in Behcet's disease. AB - Behget's disease is a systemic disease of unknown etiology with a chronic relapsing course,characterized by oral aphtous, genital ulcers, ocular lesions, and occasionally vasculitis. Major asymptomatic vascular complications should always be considered in patients with Behget'sdisease. We present the surgical treatment of two male Behget's patients of 41 and 30 years of age with ruptured infrarenal abdominal aortic aneurysms. The urgent repairs of ruptured abdominal aortic aneurysms were performed successfully in both patients. Because Behget's disease is usually seen at young ages, vascular assessment should also be done routinely for early diagnosis and therapy, which can be life-saving. PMID- 14738094 TI - Reply to: Anatomic variants associated with newborn circumcision complications. PMID- 14738095 TI - Inferior vena cava thrombus removal using hypothermic circulatory arrest in two patients with osteosarcoma. AB - In this case report, the technique of deep hypothermia and circulatory arrest (DHCA) is used to successfully extract extensive symptomatic tumor thrombus from the inferior vena cava (IVC) in two patients with osteosarcoma. This is the first description of the use of this modality for treatment of osteosarcoma involving the IVC. Osteosarcoma and the use of DHCA are discussed. PMID- 14738096 TI - Post-traumatic arteriovenous fistula presented with hypertrophic palmaris digitalis II. AB - A 19-year-old female was admitted to our department because of a hypertrophic palmaris digitalis II of the left hand displaying color changes. A thrill was easily palpated over the medial part of the finger. A review of her history revealed that this hypertrophy occurred after a blunt trauma to the left hand. Angiography showed arteriovenous fistulas between the digital branches of the radial and ulnar arteries and cephalic vein at the palmaris digitalis II of the left hand. These arteriovenous connections were closed with simple ligation under regional anesthesia. Post-operatively she had no sign of ischemia, and no thrill was palpated or auscultated. PMID- 14738097 TI - Preoperative embolization of carotid body tumors with microsphere particles. PMID- 14738098 TI - Patient adherence with hypertension medication. PMID- 14738099 TI - Terbinafine is more effective than itraconazole in treating toenail onychomycosis: results from a meta-analysis of randomized controlled trials. AB - BACKGROUND: Toenail onychomycosis is a challenge for clinicians to treat, and this challenge is compounded by conflicting information in the medical literature concerning the efficacy of the two principal agents used in its treatment: terbinafine and itraconazole. OBJECTIVE: The purpose of this meta-analysis is to compare the efficacy of terbinafinewith that of itraconazole in the treatment of toenail onychomycosis caused by dermatophytes. METHODS: A Medline search was performed for all English language publications from 1966 to June 1999 on the use of terbinafine and itraconazole in the treatment of toenail onychomycosis. Included were randomized studies in which subjects received no less than 3 months (or cycles) and no more than 4 months (or cycles) of either terbinafine or itraconazole. Data were abstracted and statistical analyses (random effects model, fixed effects model, and Peto's method) were applied. RESULTS: Thirteen studies were included from the original literature review of 1636 total referenced reports; four studies did not fulfill our inclusion or exclusion criteria. The primary analysis of six studies directly comparing terbinafine to itraconazole resulted in an odds ratio ranging from 1.8 (95% CI = 1.8, 2.8) to 2.9 (1.9, 4.1). The secondary analysis of three studies comparing either itraconazole or terbinafine to placebo estimated an odds ratio of 1.1-1.7. The former shows that terbinafine is 80%-190% more likely to result in mycologic cure than is itraconazole; the latter demonstrates a 10%-70% greater likelihood. The difference between the relative efficacies of terbinafine and itraconazole was highly statistically significant (p < 0.0001). CONCLUSION: Meta-analysis of the published worldwide literature finds that terbinafine is significantly more effective than itraconazole at achieving mycologic cure of toenail onychomycosis. PMID- 14738100 TI - Axillary granular parakeratosis. AB - BACKGROUND: Granular parakeratosis is a hyperkeratotic eruption of unknown etiology that occurs in the axilla and other intertriginous areas. The histologic features are unique for the retention of keratohyalin granules in the stratum corneum. OBJECTIVE: The aim of the article is to review the clinical manifestations and treatment of granular parakeratosis. METHODS: A case report and literature review are presented. RESULTS: Twenty-six cases of granular parakeratosis now have been reported and reviewed. CONCLUSION: Granular parakeratosis occurs predominantly in middle age women and most frequently in the axilla. A number of treatments have been applied to the 26 cases reported to date, without consistent responses. No evidence-based therapy can be suggested at present. PMID- 14738101 TI - Necrolytic migratory erythema: case report and clinical review. AB - BACKGROUND: Necrolytic Migratory Erythema (NME) is a rarely encountered dermatologic condition. It is the characteristic feature of a paraneoplastic syndrome associated with neuroendocrine pancreatic tumors. A case of NME initially diagnosed and treated as psoriasis is reviewed. A review of the current literature regarding NME is also included. OBJECTIVE: The purpose of this report is to familiarize dermatologists with a rare and potentially fatal disorder. Early recognition of NME can lead to possible cure, while delayed identification of the disease is associated with metastatic disease and a poor prognosis for the patient. CONCLUSION: When evaluating the patient who presents with a dermatitis and weight loss, it is important to exercise great caution. The differential diagnosis should be re-evaluated in a disease that is not responsive to first line therapy before further treatment is considered. PMID- 14738102 TI - Aggressive tumors of the concha: treatment with postauricular island pedicle flap. AB - BACKGROUND: The concha is a partially hidden anatomical area characterized by difficult access and reconstruction. OBJECTIVE: We describe the use of the postauricular (revolving door) island pedicle flap in the treatment of aggressive tumors of the concha. METHODS: We present two patients with aggressive cutaneous tumors localized in auricular concha. One of the patients presented with a large ulcerated basal cell carcinoma. The second patient had received a heart transplant, was receiving immunosuppressive therapy, and was affected by a squamous cell carcinoma. RESULTS: Both defects were closed using a posterior auricular island flap that was advanced through cartilage with excellent cosmetic results. CONCLUSION: The postauricular island pedicle flap is well suited for conchal reconstruction because of its proximity to the defect. The flap was transferred from posterior to anterior toward the concha, followed by primary closure of the retroauricular donor site. PMID- 14738103 TI - Traumatic intrapericardial herniation of the stomach. PMID- 14738104 TI - Pulmonary tumor embolism secondary to osteosarcoma. PMID- 14738105 TI - The risk of lethals for hypermutating bacteria in stationary phase. PMID- 14738106 TI - A doctor discusses...advance care directives. PMID- 14738107 TI - Crime and punishment and treatment: latest findings in the treatment of drug related offenders. PMID- 14738108 TI - [Eosinophilia]. PMID- 14738109 TI - Early prenatal diagnosis of familial intestinal polyatresia (FIPA) in a 19 weeks old fetus with sonographic and postmortem findings. AB - Familial intestinal polyatresia (FIPA) is a rare autosomal recessive disorder. In this article we present a new prenatally diagnosed case with FIPA from consanguineous parents with two affected daughters. The fourth pregnancy was diagnosed prenatally with FIPA at 18 weeks sonographically and these findings were confirmed by postmortem examination. PMID- 14738110 TI - Moderate hearing loss and pseudodominant inheritance due to L90P/35delG mutations in the GJB2 (connexin 26) gene. AB - Mutations in the GJB2 (connexin 26-Cx26) gene are responsible for 20-50% of cases with prelingual non-syndromic deafness in a large part of the world including Turkey. Although most of the cases with Cx26 deafness have a recessive mode of inheritance, a small group of families demonstrated dominant or pseudodominant inheritance. In this report we present a Turkish family in which the proband had congenital profound deafness and was found to be homozygous for the 35delG mutation, whereas the father and a paternal uncle who had milder, late-onset sensorineural hearing loss had compound heterozygous 35delG and L90P mutations. This family and previous reports with the L90P mutation demonstrate that the hearing loss associated with the L90P/35delG genotype is consistently milder than that of 35delG homozygotes. GJB2 gene screening should be considered in families with seemingly dominant inheritance and late-onset moderate hearing loss. PMID- 14738111 TI - A case with Pyle type metaphyseal dysplasia: clinical, radiological and histological evaluation. AB - Pyle type metaphyseal dysplasia is a rare autosomal recessive disease that is primarily affect metaphyses. Here we present a case with Pyle type metaphyseal dysplasia. The characteristic features of the case were metapyhseal broadening with undertubulation and Erlenmeyer flask sign at distal femoral and proximal tibial metaphyses. There were also platyspondyly with biconcave lens appearance of the vertebral bodies, congenital hip dislocation and normal cranium. Bone histopathology showed decreased number of osteoclasts. To the best of our knowledge, this is the first reported case of Pyle type metaphyseal dysplasia from Turkey. PMID- 14738112 TI - A low rate of trisomy 21 in twin-pregnancies: a cytogenetics retrospective study of 278 cases. AB - From January 1st 1990 until December 31st 2001, we collected 19686 prenatal diagnosis (on amniocentesis and chorius villus sampling). Five hundred twelve samples (2.6%) concerned 278 twin pregnancies. The most frequent indications were maternal age > or = 35 years (108/278 = 38.8%), medically assisted procreation (34/278 = 12.3%), positive ultrasound (20/278 = 7.2%). Chromosome abnormalities were found in eight twin-pregnancies (2.9%): five with only one fetus affected [47,XX,+ 18; 45,XX,t( 13;14); 47,XYY; 47,XXX; 45,XX, t(13;14)], two with both fetuses showing the same chromosomal abnormality [inv(11)(q21q25); 47,XX,+ 18] and one with only one fetus tested [47,XX,+ 18]. In total, we found eight autosomal abnormalities, four inherited balanced rearrangements (two robertsonian translocations and two paracentric inversions of chromosome 11) and four trisomies 18. We also observed two sex chromosome abnormalities interesting only one of the two fetuses. Surprisingly, we did no detect any Down Syndrome among this population. The frequency of Down Syndrome was significantly (p < 0.05) lower in our population of twin pregnancies (0.0%) as compared to the observed incidence in singleton pregnancies during the same period (163/19162 = 0.9%). PMID- 14738113 TI - Sporadic familial polydactyly. AB - Polydactyly is one of the most common congenital deformities of the hands. It can occur as an Isolated disorder, in association with other malformations of the hands or feet, or as part of a syndrome. It can occur sporadically but it can also be inherited with a mainly autosomal dominant inheritance. We present a family with three out of four members of the last generation with no previous history of polydactyly. Although the affected children presented different phenotypes of polydactyly, probably all three cases were the result of the same mutation seen in one of the parents. In addition no syndromic association could be found. Our experience with the management of this family's members is presented and possible etiological factors and treatment modalities are discussed. PMID- 14738114 TI - A new case of dup(1)(q21.2q12) in an individual with mild mental retardation. AB - Proximal duplications of the long arm of chromosome 1 are rare and the few patients that have been described in literature have multiple congenital abnormalities and/or mental retardation. The present paper describes the clinical and cytogenetic findings of an adult patient with only mild mental retardation and some minor malformations. The patient carries an inverted duplication of 1q12q21.2. PMID- 14738115 TI - Pregnancy outcome of 30 fetuses with cystic hygroma diagnosed during the first 15 weeks of gestation. AB - The goal of the study was to assess the prognostic value of ultrasound finding of fetal cystic hygromas. Thirty cases of septated cystic hygromas were diagnosed at 10-15 weeks gestation by transabdominal ultrasound and followed through pregnancy. The rate of abnormal karyotype was found to be 61% and the global rate of unfavorable outcome, independently of karyotype result, as high as 96%. These data suggest that cautious genetic counselling should be offered when such cystic hygromas are noticed during the first 15 weeks of gestation, even with a normal karyotype. PMID- 14738116 TI - Paracentric inversion in the short arm of chromosome 1. Report of a family and review of the literature. AB - In general, carriers of paracentric inversions are phenotypically normal, although individual reports describe like occurrence of infertility, miscarriages and mental retardation in inversion carriers. We present a family with paracentric inversion of 1p [karyotype: 46,XY/XY, inv(1)(p13.2p36.2)] in 7 of the 12 investigated family members. The index patient, a four year-old boy was referred for motor and mental retardation. The possible relationship between the paracentric inversion and the MR/MCA syndrome in the index patient of this family is briefly discussed. PMID- 14738117 TI - Second polar body inclusion results in diploid/triploid mixoploidy. AB - We report three cases with a typical diploid/triploid mixoploidy. Cytogenetic analysis showed a normal diploid karyotype in peripheral blood lymphocytes and a mixture of diploid and triploid cells in skin fibroblasts. We analysed microsatellite markers in patients blood lymphocytes and skin fibroblasts and compared the results with the microsatellite markers in the parents. The extra haploid set was in all three cases of maternal origin. In one case the markers were not very informative but in two cases pericentromeric markers showed a single dose of one paternal allele and a double dose of one maternal allele, more telomeric markers showed one paternal allele and two different maternal alleles. These observations can only be explained by the inclusion of the second polar body in one of the blastomeres at the cleavage stage. PMID- 14738118 TI - Floating-Harbor syndrome in two sisters: autosomal recessive inheritance or germinal mosaicism? AB - We describe the clinical histories and physical findings most compatible with the diagnosis of Floating-Harbor syndrome in two sisters. The genetic basis of the Floating-Harbor syndrome is still unclear, and family data are in favour of autosomal recessive inheritance although germinal mosaicism for an autosomal dominant gene mutation cannot be fully excluded. PMID- 14738119 TI - Genetic testing-genetic counseling: to keep thalassemia at bay. PMID- 14738120 TI - Double trisomy (1q32-->qter and 21pter-->q22) in a newborn female resulting from a maternal t(1;21)(q32;q22). PMID- 14738121 TI - Second-trimester diagnosis of mosaic idic(20)(p11) confined to amniocytes without an abnormal phenotype. PMID- 14738122 TI - Desquamative gingivitis: a sign of mucocutaneous disorders--a review. AB - Desquamative gingivitis is a clinical term to describe red, painful, glazed and friable gingivae which may be a manifestation of some mucocutaneous conditions such as lichen planus or the vesiculobullous disorders. It is important to be aware of this rare clinical entity so as to distinguish desquamative gingivitis from plaque induced gingivitis which is an extremely common condition, easily recognized and treated daily by the dental practitioner. This article gives an overview of desquamative gingivitis, its presentation, the possible causes, diagnosis and treatment. Early recognition of these lesions may prevent delayed diagnosis and inappropriate treatment of potentially serious dermatological diseases. PMID- 14738123 TI - Dental implants: maintenance, care and treatment of peri-implant infection. AB - Osseointegration is becoming increasingly routine in the rehabilitation of partially or fully edentulous patients. However, the surrounding tissues may be subject to inflammatory conditions similar to periodontal disease and so require maintenance. This article discusses the background, aetiology, diagnosis of peri implant diseases, and the maintenance, care and treatment of peri-implant infection in osseointegrated implants. Three case studies are presented to illustrate points in the care of implants. PMID- 14738124 TI - Clinicians' choices of restorative materials for children. AB - BACKGROUND: Recently, there has been an expansion in the range of tooth-coloured restorative materials available. In 1999, the National Health and Medical Research Council recommended clinicians use alternatives to amalgam in children 'where appropriate'. METHODS: A three-part 29-item questionnaire was developed, tested in a focus group, and distributed to members of the Australasian Academy of Paediatric Dentistry (AA; paediatric dentists and paediatric dentistry postgraduate students; n=55), and the Australian and New Zealand Society of Paediatric Dentistry, Victorian Branch (SPD; general dentists and dental therapists; n=50). Participant information, material choices, and six hypothetical clinical scenarios were addressed. RESULTS: The overall response rate was 74 per cent. For both groups, the first ranked factor influencing choice of restorative material for vital primary teeth was child age, and caries experience for vital first permanent molars. For moderate-sized Class I and II restorations in primary molars, a tooth-coloured material was chosen by 92 and 84 per cent respondents respectively. For restoring two separate proximal lesions in a primary molar, 65 per cent chose a tooth-coloured material followed by a stainless steel crown (27 per cent; all AA members), then amalgam (8 per cent). The SPD respondents were significantly more likely to choose glass ionomer cement for Class I and II restorations and for restoring two proximal lesions (all p=0.000) in primary molars than AA respondents, who were more likely to choose composite resins/compomers or amalgam/stainless steel crowns for these restorations. Younger respondents (21-40 years) were significantly more likely to choose composite resins/compomers or amalgam/stainless steel crowns (p=0.048) than older respondents (41-65 years), who were likely to choose glass ionomer cement. CONCLUSIONS: For Class I and II restorations in primary molars, glass ionomer cement was the material chosen most frequently (SPD respondents); preference for amalgam or stainless steel crowns was low (both SPD and AA groups). The wide range of materials chosen for the hypothetical clinical scenarios suggests the need for guidelines on selection of restorative materials, and the need for longitudinal studies to follow actual clinical outcomes of the materials chosen. PMID- 14738125 TI - Precision ablation of dental enamel using a subpicosecond pulsed laser. AB - In this study we report the use of ultra-short-pulsed near-infrared lasers for precision laser ablation of freshly extracted human teeth. The laser wavelength was approximately 800nm, with pulsewidths of 95 and 150fs, and pulse repetition rates of 1kHz. The laser beam was focused to an approximate diameter of 50microm and was scanned over the tooth surface. The rise in the intrapulpal temperature was monitored by embedded thermocouples, and was shown to remain below 5 degrees C when the tooth was air-cooled during laser treatment. The surface preparation of the ablated teeth, observed by optical and electron microscopy, showed no apparent cracking or heat effects, and the hardness and Raman spectra of the laser-treated enamel were not distinguishable from those of native enamel. This study indicates the potential for ultra-short-pulsed lasers to effect precision ablation of dental enamel. PMID- 14738126 TI - Remineralization of enamel subsurface lesions in situ by sugar-free lozenges containing casein phosphopeptide-amorphous calcium phosphate. AB - BACKGROUND: The anticariogenic potential of casein phosphopeptide-amorphous calcium phosphate nanocomplexes (CPP-ACP) has been demonstrated using laboratory, animal and human in situ caries models. The aim of this study was to determine the effect of CPP-ACP incorporation into a sugar-free lozenge (pressed mint tablet) on enamel remineralization in a human in situ model. METHODS: The study utilized a double-blind, randomized, cross-over design with four treatments: (i) a lozenge containing 56.4mg (3 per cent w/w) CPP-ACP; (ii) a lozenge containing 18.8mg (1 per cent w/w) CPP-ACP; (iii) a lozenge not containing CPP-ACP; and (iv) a no lozenge nil-treatment control. Ten subjects wore removable palatal appliances with four, human-enamel, half-slab insets containing subsurface lesions. Lozenges were consumed, without chewing, four times per day for 14 days duration. After each treatment period the enamel slabs were removed, paired with their respective demineralized control, embedded, sectioned and subjected to microradiography and computer-assisted densitometric image analysis to determine the level of remineralization. RESULTS: The incorporation of CPP-ACP into the lozenge significantly increased enamel subsurface lesion remineralization with 18.8 and 56.4mg of CPP-ACP increasing remineralization by 78 and 176 per cent respectively, relative to the control sugarfree lozenge. CONCLUSION: This study demonstrates that lozenges are a suitable vehicle for the delivery of CPP-ACP to promote enamel remineralization. PMID- 14738127 TI - Efficacy of cardiopulmonary resuscitation performed in a dental chair. AB - BACKGROUND: Within the dental setting, historically there has been some concern as to whether cardiopulmonary resuscitation (CPR) can be performed effectively in the dental chair. This study tested the hypothesis that there is no difference in the efficacy of CPR performed in the dental chair or on the floor. METHODS: Four cycles of two-person CPR were performed by three health professionals on a manikin positioned alternately on the floor and in a dental chair. Ventilation was performed using a Laerdal pocket mask, without oxygen supplementation. Compression and ventilation performance was recorded using a computerized manikin skill meter. RESULTS: Each of the participants was able to achieve a mean cardiac compression depth of between 41 and 50cm, irrespective of the CPR surface. The only statistically significant difference found in expired air resuscitation (EAR) and external cardiac compression performance was that 37 per cent of ventilations performed on the floor were deemed to be too shallow, compared to only 15 per cent in the dental chair (p=0.001). CONCLUSIONS: It is possible for those trained in basic life support to perform CPR effectively in the dental chair. Each of the participants agreed that CPR, in particular EAR, was easier to perform when the manikin was in the dental chair compared with the floor. Dentists are encourage to regularly update their CPR knowledge and skills, including the practice of CPR in the dental chair. PMID- 14738128 TI - Microscopic investigation of artificially demineralized surface enamel exposed to controlled intra-oral periods. AB - BACKGROUND: Arrest and regression of enamel caries is generally followed by clinical surface changes in enamel. The aim of this study was to evaluate microscopic surface changes in demineralized enamel submitted to toothbrushing with a fluoridated dentifrice or to a fluoridated gel in situ. METHODS: Enamel blocks demineralized by an acidic gel, were built in plates of five volunteers during three periods of 14 days each: exposure to the oral cavity (C+); brushing with a fluoridated dentifrice three times a days (B); application of a 2 per cent sodium fluoride gel and exposure to the oral cavity (F). Eight blocks were kept in 100 per cent humidity under refrigeration as negative controls (C-). RESULTS: Stereomicroscopic examination revealed that 15, 47.4 and 30 per cent of samples from C+, B and F presented enamel cavitation after the experimental period. The demineralized enamel area in these groups decreased considerably compared to C-, both in cavitated and non-cavitated samples, although it was statistically significant only in cavitated samples from B (p=0.003; t test). Scanning electron microscopy revealed that C+ and F retained irregularities caused by the acid challenge, while B presented a more leveled profile of the enamel surface. CONCLUSIONS: Exposure of demineralized enamel to intra-oral periods reduced the demineralized enamel area. Tooth brushing enhanced this reduction, because of surface abrasion. PMID- 14738129 TI - Non-surgical management of an oro-antral fistula in a patient with HIV infection. AB - BACKGROUND: The risk of post-extraction complications is higher in patients who are immunosuppressed compared to other patients with normal immune function. In addition, invasive dental procedures are more likely to have serious complications in these patients. This case report demonstrates an effective non surgical procedure to treat an oro-antral fistula in an HIV-infected man. METHODS: The oro-antral fistula was de-epithelialized under local anaesthesia and the patient wore a surgical splint continuously, removing it only for cleaning, for an eight week period. Chlorhexidine gel was regularly applied to the fitting surface of the splint and the oro-antral communication. The patient was reviewed on a regular basis. RESULTS: This procedure resulted in resolution of the patient's symptoms within two weeks. Complete healing of the oro-antral fistula was evident following eight weeks of wearing the surgical splint. CONCLUSIONS: This procedure provided an effective method of treating an oro-antral fistula in an immunocompromised patient without causing any detrimental effects to the patient's overall health. Adequate pre-surgical assessment of patients prior to extractions is important in all patients to help prevent the occurrence of such complications. PMID- 14738130 TI - A model for the prediction of tooth wear in individuals. AB - BACKGROUND: Easily applied, age-specific standards are useful in assisting clinicians to decide whether the extent of tooth wear observed in a given patient at a specific age is acceptable or whether intervention is indicated. METHOD: A simple method of scoring tooth wear and previously established mathematical relationships between tooth wear scores and age, provide the basis of a method for predicting tooth wear scores. In its most specific form, the method can be used to predict tooth wear scores for individual patients at subsequent ages. Alternatively, tables or graphs can be consulted to provide a less precise prediction of tooth wear that can assist clinicians to decide whether the amount of wear reflects high, moderate or low rates of wear at the specified age. CONCLUSION: In cases where there is no reason to believe that aetiological factors have changed significantly over time, data obtained from the methods described can assist clinicians to plan appropriate management for patients presenting with unacceptable levels of tooth wear. PMID- 14738131 TI - Destructive nocturnal bruxing. PMID- 14738132 TI - Minor oral surgical procedures in patients on oral anticoagulants. PMID- 14738133 TI - Dental extractions and anticoagulants. PMID- 14738134 TI - Bisphosphonates and avascular necrosis of the jaws. PMID- 14738135 TI - Multicolor karyotyping in acute myeloid leukemia. AB - Cytogenetic data have significantly contributed to our understanding of the heterogeneity of acute myeloid leukemia (AML). In AML, numerous recurrent chromosomal aberrations have been identified, and several of them, e.g. t(8;21)(q22;q22), t(15;17)(q22;q11-12), inv(16)(p13q22), are specific for distinct subgroups. Furthermore, chromosomal aberrations have proved to be of paramount prognostic importance for remission induction and survival. Chromosome analysis using classical cytogenetic banding techniques often fails to completely resolve complex karyotypes and cryptic translocations not identifiable by these techniques have been detected using molecular cytogenetic methods. While fluorescence in situ hybridization (FISH) has become an indispensable tool for screening and follow-up of known aberrations, the techniques of spectral karyotyping (SKY) and multiplex-fluorescence in situ hybridization (M-FISH) allow for the simultaneous visualization of all chromosomes of a metaphase in a single hybridization step, and thereby enable screening for the aberrations present without their prior knowledge. Therefore, with the introduction of these techniques in 1996 the comprehensive analysis of complex karyotypes and the identification of new, hitherto cryptic translocations and, ultimately, the identification of new disease subgroups seemed possible. Since, more than 600 cases of AML and MDS have been analyzed. Herein, we attempt to summarize the data published and discuss what has been achieved towards realization of these goals. PMID- 14738136 TI - The role of DNA hypermethylation in the pathogenesis and prognosis of acute lymphoblastic leukemia. AB - The hallmark of acute lymphoblastic leukemia (ALL) is a progressive appearance of malignant cell behavior that is triggered by the evolution of altered gene function. ALL has traditionally been viewed as a genetic disease, however, epigenetic defects also play an important role. DNA promoter methylation has gained increasing recognition as an important mechanism for transcriptional silencing of cancer related genes. The hypermethylation-associated inactivation affects virtually all of the pathways in the ALL cellular network, such as the cell cycle, apoptosis and adhesion. The identification of these methylation abnormalities and elucidation of the mechanistic events surrounding them are of prime importance, as the methylation status of ALL cells can be used as prognostic biomarker and also can be manipulated in vivo with demethylating agents. PMID- 14738137 TI - Protein kinases in the regulation of apoptosis in B-cell chronic lymphocytic leukemia. AB - The involvement of several protein kinase pathways in the regulation of apoptosis and cell survival has been analyzed in a wide range of models. This article reviews current understanding of the protein kinases involved in the control of apoptosis in B-cell chronic lymphocytic leukemia (B-CLL) cells. Protein kinase C (PKC), phosphatidylinositol 3-kinase (P13K) and nuclear factor-kappa B (NF kappaB) play important roles in the survival of these leukemic cells. These survival pathways affect proteins involved in the control of apoptosis by altering their expression or function. The elucidation of the signal transduction network involved in the survival of B-CLL cells could provide novel pharmacological targets for the therapy of B-CLL. PMID- 14738138 TI - Depletion of host reactive T cells by photodynamic cell purging and prevention of graft versus host disease. AB - Graft versus Host Disease (GVHD) is the principal cause of morbidity and mortality in patients undergoing allogeneic stem cell transplant. T cell depletion has been recognized as a method of reducing the incidence of GVHD in allogeneic transplants. Until recently, most T cell depletion methods were non selective in reducing lymphocytes. Rhodamine purging is one method, which selectively reduces alloreactive T cells preventing GVHD. We review here the methods of non-selective and selective T cell depletion, particularly the newer method of photodynamic purging utilizing rhodamine. PMID- 14738139 TI - Hematopoiesis and retinoids: development and disease. AB - Retinoids function as activating ligands for a class of nuclear receptors that control gene expression programs for a wide range of tissues and organs during embryogenesis and throughout life. Over the years, three sets of observations have spurred interest in the function of retinoids with respect to development and disease of hematopoietic cells. Since the 1920s, epidemiological studies indicated altered hematopoiesis in vitamin A-deficient (VAD) human populations. More recently, the ability of retinoids to affect various aspects of hematopoietic development has been demonstrated in vitro. Finally, it was discovered that the gene encoding a retinoid receptor is a key target for chromosomal translocations that cause acute promyelocytic leukemia (APL). More recent investigations using targeted gene disruptions, VAD animal models, and mouse models of leukemia have continued to shed light on the function of the retinoid pathway in blood cells. It is now clear that retinoids are required for normal hematopoiesis during both yolk sac and fetal liver stages of hematopoiesis, while the pathway has at least modulatory functions for bone marrow derived progenitors. Studies of normal development and APL have provided complementary insight into the molecular control of blood cell differentiation. Here we review the evidence for retinoid requirements in hematopoiesis and also summarize current ideas regarding how this pathway is subverted in leukemia. PMID- 14738140 TI - cAMP/PKA-mediated regulation of erythropoiesis. AB - The role of cyclic AMP (cAMP) as second messenger in erythropoiesis has been suggested in the early 1980s. However, careful analysis showed that cAMP is not generated in direct response to the main erythropoiesis-controlling cytokines such as erythropoietin (Epo). As a result, cAMP disappeared from the central stage in research of erythropoiesis. Instead, other signal transduction pathways, including the Ras/extracellular regulated kinase (ERK)-pathway, the phosphatidylinositol 3-kinase (P13K) and the signal transducer and activator of transcription (STAT5)-pathways, have been found and explored. In concert, these signaling pathways control the transcriptional machinery of erythroid cells. Although cAMP is not directly generated in response to Epo stimulation, it has recently been demonstrated that increased cAMP-levels and in particular the cAMP dependent protein kinase A (PKA) can modulate erythroid signal transduction pathways. In some cases, like the ERK-signaling pathway, PKA affects signal transduction by regulating the balance between specific phosphatases and kinases. In other cases, such as the STAT5 pathway, PKA enhances Epo signaling by inducing recruitment of additional co-regulators of transcription. In addition to STAT5, PKA also activates other transcription factors that are required for erythroid gene expression. This review discusses the impact of cAMP/PKA on Epo-mediated signaling pathways and summarizes the role of cAMP in malignant erythropoiesis. PMID- 14738141 TI - Diffuse large B-cell lymphoma arising in nodular lymphocyte predominant hodgkin lymphoma. A report of 21 cases from the Nebraska Lymphoma Study Group. AB - We sought to investigate the clinical characteristics and pathologic features and survival outcome of patients with diffuse large B-cell lymphoma (DLBCL) arising in nodular lymphocyte predominant Hodgkin's disease (NLPHL), since controversy regarding their prognosis exists in the literature. Twenty-one patients with DLBCL arising either concurrently with (n = 7) or subsequent to (n = 14) a diagnosis of NLPHL were identified in the Nebraska Lymphoma Study Group Registry. The clinical and pathologic features of the cases were evaluated, and survival analysis was performed from the time of diagnosis of DLBCL. The median time to the development of DLBCL in those with prior NLPHL was only one year (range, 0.5 24 years). The median age of the patients at the time of diagnosis of DLBCL was 46 years (range, 18-72 years) and the male to female ratio was 17:4. Ten patients presented with nodal DLBCL only, six patients presented with both nodal and extranodal involvement, and five patients presented with only extranodal DLBCL. Eleven patients had limited stage (I/II) disease and 10 had advanced stage (III/IV) disease. The median overall survival (OS) and failure-free survival (FFS) of the entire group was 35 months and 11 months, respectively, and the predicted five-year OS and FFS was 31 and 18%, respectively. There were no significant differences in the survival outcomes between patients with DLBCL arising in NLPHL and age- and sex- matched patients with de novo DLBCL. In conclusion, our findings suggest that patients with DLBCL arising in NLPHL have a prognosis similar to those with de novo DLBCL and should be treated aggressively. PMID- 14738142 TI - The role of anthracyclines in combination chemotherapy for the treatment of follicular lymphoma: retrospective study of the Intergruppo Italiano Linfomi on 761 cases. AB - In order to elucidate the role of anthracycline based combination chemotherapy regimens for the treatment of follicular lymphoma we conducted a retrospective study on a large series of patients with a histologically confirmed diagnosis of follicular lymphoma. The Italian lymphoma intergroup (ILI) promoted a retrospective study of patients with follicular lymphoma treated in cooperative trials between 1985 and 1996. Six hundred and thirty three cases were treated with an anthracycline-containing regimen and 128 patients were treated without anthracyclines. The two groups were prognostically comparable; in particular, no difference was observed according to both IPI and ILI prognostic index. Results showed a complete remission (CR) rate for patients treated with anthracyclines was 69.2% and overall response rate was 92.5%. After a median follow-up of 51 months (54 months for patients still alive), the 5- and 10-year overall survival (OS) rates were 80 and 66%, respectively. Disease-free survival (DFS) and failure free survival (FFS) rates at 5 years were 61 and 49%, respectively. In the group of patients treated with combination chemotherapy not including anthracyclines, the CR rate was 67.5% and the overall response rate was 85.4%. A longer OS (80% at 5 years) was observed in patients treated with anthracyclines compared to 67% OS rate in patients treated without anthracyclines (p = 0.0004). FFS was significantly longer in patients treated with anthracyclines (49 vs. 34% p = 0.006). Patients treated with anthracyclines with low or intermediate risk according to ILI prognostic index showed a significantly longer OS (p = 0.0001 andp = 0.0009, respectively); those in the high-risk group showed a trend for a longer survival. In conclusion, this retrospective study shows that patients with follicular lymphoma treated with an anthracycline containing regimen had a better outcome compared to patients treated with other combination regimens non including anthracyclines in terms of CRs, OS and FFS. On the basis of these results anthracycline-containing regimens (ACR) should be considered as the standard treatment of patients with advanced follicular lymphoma. PMID- 14738143 TI - Efficacy of vinblastine, bleomycin, methotrexate (VBM) combination chemotherapy with involved field radiotherapy in early stage (I-IIA) Hodgkin disease patients. AB - Vinblastine, bleomycin, methotrexate (VBM) combination chemotherapy (CT) with involved field radiotherapy (IFRT) was first described by the Stanford group as an active regimen in early stage Hodgkin's disease (HD). Here, we report our retrospective experience of a modified VBM schedule + IFRT in a similar group of patients. From 1988, 49 patients with stage I-IIA HD received vinblastine (VBL) 6 mg/m2, bleomycin (BLM) 10 IU/m2, methotrexate (MTX) 30 mg/m2 day 1,8 every four weeks for three cycles; IFRT was delivered four weeks later followed by three additional cycles of VBM with a dose reduction of BLM (6 IU/m2). The regimen was well tolerated, with grade 3-4 neutropenia occurring in 20 patients. No acute or late pulmonary toxicity was recorded in our series. Estimated Freedom from Progression (FFP) and Overall Survival (OS) at five years are 75% (95% CI, 60.1% 92.2%) and 85% (95% CI, 73.6%-98.1%), respectively. In this retrospective analysis, VBM + IFRT treatment with bleomycin dose reduction seems safe and active. Such combination could be considered as first line treatment for early stage HD patients with favorable prognosis and/or not suitable for anthracyclines containing regimens. PMID- 14738144 TI - Post-traumatic stress disorder and quality of life in long-term survivors of Hodgkin's disease and non-Hodgkin's lymphoma in Israel. AB - Post-traumatic stress disorder (PTSD) has not been examined systematically in long-term survivors of lymphoma. In this study, PTSD and health related quality of life (HRQoL) were assessed in 44 patients with Hodgkin's disease (n = 8) or non-Hodgkin's lymphoma (n = 36). Forty-four individuals who had experienced traumatic events as defined by the Diagnostic and Statistical Manual-IV (DSM-IV) as possible triggers for PTSD served as controls. The study participants were administered two questionnaires-the PTSD inventory scale and the Short Form-36 (SF-36) HRQoL instrument measuring physical and mental HRQoL. Full PTSD was defined as meeting the DSM-IV criteria for the diagnosis in all three symptom groups measured on the PTSD inventory scale-intrusion, avoidance and hyper arousal and partial PTSD as meeting the diagnostic criteria in two of the symptom groups. There was a significant increase in the hyper-arousal scale in the lymphoma survivor group (F 5, P < 0.05). Overall, full or partial PTSD was found in 14 lymphoma survivors (32%) and in 11 individuals (25%) in the control group (difference not significant). Survivors whose disease had started at an earlier age suffered significantly more intensive intrusion and avoidance symptoms. The lymphoma survivor group had a significantly lower physical HRQoL than the control group independent of PTSD symptoms. In both groups, the presence of PTSD symptoms correlated with a lower HRQoL. These results suggest that lymphoma is a trauma similar to other more accepted definitions of trauma which can lead to PTSD, and is associated with more severe hyper-arousal symptoms. Psychological interventions in the early stages of treatment or follow-up may help reduce the morbidity from PTSD and improve quality of life. PMID- 14738145 TI - Mutation analysis of the Nijmegen breakage syndrome gene (NBS1) in nineteen patients with acute myeloid leukemia with complex karyotypes. AB - The chromosomal instability disorder Nijmegen Breakage Syndrome (NBS) is caused by germ line mutations in the NBS1 gene. It is associated with immune deficiency, cellular hypersensitivity to ionizing radiation, and high susceptibility to lymphoid malignancies due to a defect in DNA double strand break repair. Since genetic instability has been discussed as a cause in acute myeloid leukemia (AML) with complex chromosomal aberrations, mutations in the NBS1 gene might be found in this AML subgroup. In this study, we analyzed 19 patients with AML and complex chromosomal aberrations for mutations in the NBS1 gene. Tumor DNA was analyzed by dHPLC analysis and all amplicons showing shifts were directly sequenced. One sample was found to be heterozygous for a novel 5 bp deletion in intron 12 (IVS12 53del5). By RT-PCR analysis the expected transcript and an additional faint product with skipped exon 13 was observed, indicative of aberrant splicing. This exon codes for part of the binding site of the NBS1 gene product, nibrin, to MRE11. However, we also found that all controls showed this phenomenon. Thus, the IVS12-53del5 is not responsible for the skipping of exon 13 and most probably represents a rare polymorphism. We found no further NBS1 mutations among the AML samples. Although the number of the analyzed samples is small, our study indicates that NBS1 mutations are not common in AML with a complex karyotype. PMID- 14738146 TI - Aberrant expression of HOXA9, DEK, CBL and CSF1R in acute myeloid leukemia. AB - Previous gene function analyses have indicated that HOXA9, DEK, CBL and CSF1R are aberrantly expressed in acute myeloid leukemia (AML). We analyzed the expression of these genes in a series of 41 adult patients with AML using quantitative real time RT-PCR, and tested the association of the expression with the following hematologic and clinical parameters: age, FAB, immunophenotype and karyotype aberrations. A high proportion of the patients showed over- or underexpression of the analyzed genes. DEK was overexpressed in 98% of the cases, whereas CBL, CSF1R and HOXA9 were either overexpressed in 20%, 17% and 78% or underexpressed in 20%, 42% and 15% of the cases, respectively. Patients whose karyotype contained t(8;21)(q22;q22), showed lower relative expression of HOXA9 at a statistically significant level (p < 0.05). Bone marrow samples without expression of CD34 antigen were associated with either overexpression of DEK or HOXA9. Furthermore, an association was found between the AML-M2 subtype and lower expression of CBL, CSF1R or HOXA9, and between the AML-M5 subtype and CBL or CSF1R overexpression. PMID- 14738147 TI - Low dose thalidomide in patients with relapsed or refractory multiple myeloma. AB - Remarkable results of the treatment of refractory multiple myeloma with thalidomide have been reported. In most preceding studies, the given thalidomide dose was escalated to a maximum tolerated dose of up to 800 mg/d. The frequency of adverse effects correlates with dose intensity. Since a significant gain of therapeutic effects could not be observed as thalidomide dosage was escalated, the optimal dose of thalidomide remains to be determined. We report the results of a study with low dose thalidomide (median administered dose 100 mg/d, range 50 400 mg/d). Twenty-four relapsed (n = 19) or resistant (n = 5) multiple myeloma patients were included in the study. Twelve patients (50%) received thalidomide as monotherapy, 8 patients (33%) received a combination of thalidomide and dexamethasone (every 4 weeks 40 mg/day for 4 days) and 4 patients (17%) who were resistant to vincristine, doxorubicin, dexamethasone (VAD) received VAD combined with thalidomide. Overall, a response was observed in 12 patients (50%). Of the 12 patients treated with low dose thalidomide alone 5 (42%) responded, of the 8 patients who received a combination of thalidomide and dexamethasone 5 (63%) responded and of the 4 patients who had thalidomide in addition to VAD 2 patients (50%) responded. In 3 patients, thalidomide treatment had to be discontinued because of side effects and 1 patient died before response could be assessed. We conclude that low dose thalidomide is an effective and safe rescue therapy in relapsing or refractory multiple myeloma. Response to thalidomide might be dependent on prognostic parameters and tumor burden. To answer these questions larger prospective studies are necessary. PMID- 14738148 TI - Conventional dose fludarabine-based regimens are effective but have excessive toxicity in elderly patients with refractory chronic lymphocytic leukemia. AB - The best approach to elderly patients with relapsing chronic lymphocytic leukemia (CLL) or disease refractory to conventional therapy with alkylating agents has not yet been established. Fludarabine and its combination with mitoxantrone and/or cyclophosphamide, which is the most effective treatment in younger patients, has not been extensively utilized in the elderly CLL. Here we report our results with fludarabine-based chemotherapy in 32 previously treated patients over the age of 65 years. The overall response rate was 59% with no complete remission, 3 nodular partial remissions and 16 partial remissions. The median time to progression of disease was 7 months. Only 10 patients completed the entire treatment program, because of poor compliance due to toxicity. Eight patients developed neutropenic fever, 14 severe bacterial infections and 2 patients showed progressive encephalopathy. For comparison, in a younger group of patients with refractory CLL (< 65 years), 38 of 50 patients completed the treatment plan, and the ORR was 80% (10 CR, 11 PR-nodular, 19 PR) with a median response of 12 months. Neutropenic fever was diagnosed in 10 and severe bacterial infection in 4 patients. In conclusion, fludarabine-based chemotherapy is effective for refractory CLL, however, excessive toxicity such as severe infections and neurological complications, do not allow completion of treatment in the majority of the elderly patients. Because maintenance of a good quality of life should be the main goal in the elderly CLL population, dose reduction of fludarabine and the appropriate use of myeloid growth factors and prophylactic antibiotics appear mandatory in this group of patients. PMID- 14738149 TI - Anti-CD20 therapy for chronic lymphocytic leukemia-associated autoimmune diseases. AB - Rituximab is active in chronic lymphocytic leukemia (CLL) and may interfere with autoantibodies production in some immune diseases. We report the results of rituximab treatment in 7 patients with CLL-associated symptomatic autoimmune diseases refractory to standard immunosuppressive therapies: warm antibody hemolytic anemia (AHA) 4 patients, cold agglutinin disease (CAD) 1, immune thrombocytopenia (IT) 1, axonal degenerating neuropathy (ADN) 1. Rituximab was given at the dose of 375 mg/m2 per week for 4 weeks. One patient with AHA and one with CAD achieved complete normalization of hemoglobin levels and laboratory signs of haemolysis, with response duration (RD) of 8+ and 38+ months, respectively. In the patient with IT, complete remission was reached after the first week of treatment and RD was 6 months. The patient with ADN achieved a marked neurological improvement after rituximab therapy, with RD of 12 months. Retreatment of both patients with IT and ADN was effective. Rituximab may be an alternative agent for the treatment CLL-associated autoimmune diseases. PMID- 14738150 TI - Appraisal of the MTT-based assay as a useful tool for predicting drug chemosensitivity in leukemia. AB - The MTT-based assay relies upon the cellular reduction of tetrazolium salts to their intensely colored formazans. The test is easy to perform in hematological malignancies and is adaptable for high throughput of samples, although there are some minor limitations in its application resulting from metabolic interference. This class of assays are highly accurate for predicting drug resistance, whereas their predictive value for drug sensitivity depends on the type of disease and drug or drug combination used. They have been found to predict clinical response to fludarabine FLD in B-CLL and were useful for predetermining clinical potential of a single drug or drug combination in AML patients. Extensive studies with ALL patients have supported their advantage for selecting effective drug treatment of the disease. To conclude, pretreatment chemosensitivity assays may help in the selection of chemotherapeutic drugs with the greatest likelihood for clinical effectiveness, and in the exclusion of uneffective therapy. This can lead to improved disease management, response, survival and use of financial resources. PMID- 14738151 TI - Effects of preactivated autologous T lymphocytes on CD80, CD86 and CD95 expression by chronic lymphocytic leukemia B cells. AB - Profound immune dysfunction is a constant feature in B-cell chronic lymphocytic leukemia (B-CLL) patients. Immunological abnormalities include hypogammaglobulinemia, impaired immunoglobulin class switching and diminished germinal center formation. This state of immune suppression renders B-CLL patients highly susceptible to infections, which contribute greatly to morbidity and mortality in this disease. Impaired T cell function in B-CLL is well documented and has been suggested to result from inhibitory effects exerted by malignant B lymphocytes. Because the presence of leukemic cells may represent a major obstacle to efficient T cell activation, T lymphocytes were separated from CLL B cells, stimulated with phorbol 12-myristate 13-acetate (PMA) and ionomycin for 4h, and then cocultured with autologous leukemic B cells both at a 1:1 ratio or at the same ratio as in vivo for 24-40 h. CLL B cell expression of CD86 and CD95 was markedly upregulated using this approach, whereas CD80 expression was augmented only in a minority of patients; these effects were partially preserved even when preactivated T cells were rechallenged with CLL B cells at the same low T/B cell ratio as that observed in vivo. Finally, CD80 upregulation on CLL B cells appeared to be mainly dependent on CD40L-mediated stimulation, whereas CD86 and CD95 expression was efficiently augmented by soluble factors released by preactivated T lymphocytes. In conclusion, efficient activation of T lymphocytes in B-CLL may be achieved which, in turn, may result in enhanced antigen presenting capacity and susceptibility to apoptosis of leukemic cells via CD86 and CD95 upregulation, respectively. PMID- 14738152 TI - Murine bone marrow stromal cells: implications for their use in gene modified cell therapy. AB - The objectives of this study were to demonstrate that bone marrow stromal cells (BMSC) can be an attractive novel target of genetic modification in gene and cell therapy of hematologic disease. Here, we investigated the therapeutic effects of gene modified BMSC using a murine lymphoma model. BMSC of Balb/c AnN mice were encoded with the human IL-2 gene (hIL-2) using an adenoviral vector. About 1 x 10(6) cells of a murine B lymphoma (A20) cell line, were injected into the mice via tail vein. One week after injection of A20 cells, the mice were divided into 4 groups for BMSC therapy: No BMSC (control), unmodified BMSC, BMSC with Ad/deltaE1, and BMSC with Ad/hIL-2. Mice were observed for 8 weeks. The results demonstrated that all mice treated with BMSC (Ad/hIL-2) survived with no evidence of disease during this period of observation. All mice treated with unmodified BMSC or BMSC (Ad/deltaE1) as well as the controls developed disseminated lymphoma, and 80% of mice survived less than 4 weeks. In conclusion, the IL-2 gene-modified stromal cell is a promising therapeutic tool for murine lymphoma. PMID- 14738153 TI - Cellular pharmacology of P-ethoxy antisense oligonucleotides targeted to Bcl-2 in a follicular lymphoma cell line. AB - A P-ethoxy oligonucleotide (oligo), 20 bases long and specific for the translation initiation site of human Bcl-2 mRNA, was incorporated into liposomes to increase its intracellular delivery. This oligo selectively inhibited Bcl-2 protein expression and induced growth inhibition in t(14;18)-positive transformed follicular lymphoma (FL) cell lines. We studied the inhibitory effects of shorter liposomal P-ethoxy oligos (7, 9, 11 or 15 mer) in order to determine the activity of different oligo chain lengths targeted to the same Bcl-2 mRNA. At 12 microM, all the oligos inhibited the growth of a FL cell line. We compared the 7-mer oligo with the 20-mer oligo. The two oligos inhibited Bcl-2 protein expression similarly: 66% and 60% for the 7- and 20-mer, respectively. The uptake and retention of both oligos were also very similar. Our results indicate that the Bcl-2 inhibitory activity is maintained with P-ethoxy antisense oligos ranging from 7 to 20 bases. PMID- 14738154 TI - Aminopeptidase inhibitors inhibit proliferation and induce apoptosis of K562 and STI571-resistant K562 cell lines through the MAPK and GSK-3beta pathways. AB - A tyrosine kinase inhibitor, STI571, has been demonstrated to be effective for the treatment of chronic myelogenous leukemia (CML). STI571 inhibits tyrosine kinase activity of ABL and induces apoptosis of CML cells. However, drug resistance develops commonly in patients with blast phase CML, and has become a significant therapeutic problem. We examined the effects of aminopeptidase inhibitors on CML cell line (K562) and a STI571-resistant subline of K562. Ubenimex and the more potent aminopeptidase inhibitor, actinonin, inhibited proliferation of both K562 cells and STI571-resistant K562 cells and also induced their apoptosis in dose- and time-dependent manners. Ubenimex and actinonin induced the activation of caspase-3, and the induction of apoptosis was inhibited by pan-caspase inhibitor, indicating this apoptosis is caspase-dependent. We found that serine phosphorylation of both MAPK and glycogen synthase kinase-3beta were suppressed by aminopeptidase inhibitors in parent K562 and STI571-resistant K562 cells. The expression level of cyclin D1 protein was also reduced by ubenimex and actinonin in both cell lines. These results indicated STI571 resistance does not confer the cross-resistance to aminopeptidase inhibitors in K562 cells and revealed the new findings of aminopeptidase inhibitor-induced intracellular signaling pathways. PMID- 14738155 TI - A rare case of intravascular lymphoma diagnosed on bone marrow trephine. AB - Intravascular lymphoma (IVL) is an extremely rare form of extra-nodal non Hodgkins lymphoma characterised by the proliferation of neoplastic lymphocytes within the lumina of small arteries, veins and capillaries. The great majority of reported cases appear to be of B cell lineage. There is a wide variation in clinical presentation, and multiple organs are usually affected. We report a case of a 67-year-old man who presented with constitutional symptoms and neurological deficit and was diagnosed following bone-marrow trephine. His disease responded to polychemotherapy treatment but he died 15 months after diagnosis. This case in unusual in that it is generally felt that bone marrow is relatively spared until late in the disease and is often not clearly demonstrable histologically. In addition, this case supports the limited data that responses can be obtained following polychemotherapy treatment, although the prognosis remains generally poor. PMID- 14738156 TI - Chronic myelomonocytic leukemia coexisting with B-cell chronic lymphocytic leukemia. AB - Coexistence of B-cell chronic lymphocytic leukemia (B-CLL) and chronic myelomonocytic leukemia (CMML) is an unusal event, and to our knowledge, only four such cases have been reported in the literature. We report a 68-year-old white woman in whom these two diseases were diagnosed concomitantly. The diagnosis was made on the basis of peripheral blood count, morphology and immunophenotyping, and bone marrow cytology and histology. Interphase FISH analysis detected a 13q14.3 deletion in lymphocytes nuclei and no such abnormality in monocytes nuclei. The PCR analysis of IgH gene rearrangement in the bone marrow, as well as the peripheral blood lymphocytes, showed two different monoclonal IgH configurations as the result of biallelic clonal rearrangement of IgH genes suggesting an origin of lymphocytes from B-cell progenitors. The patient was originally treated with prednisone 1 mg/kg/day because of progressive significant thrombocytopenia, without improvement. Subsequently, she received one course of cladribine (2-CdA). Significant reduction of lymphocytes in the peripheral blood was observed. However, rapid increase of monocytes was seen shortly after the 2-CdA treatment. Subsequently, she received hydroxyurea (1.5 g/day) without hematological improvement. The patient died in January 2003, three months after diagnosis because of progression of both leukemias and associated pneumonia. Possible etiopathogenic relationship between both disorders is discussed. PMID- 14738157 TI - Genital ulcers during treatment with ALL-trans retinoic acid for acute promyelocytic leukemia. AB - Scrotal ulcer is a unique adverse effect of all-trans retinoic acid (ATRA) in patients with acute promyelocytic leukemia (APL). The pathogenesis of scrotal ulceration remains unknown. We describe genital ulcers that developed in four patients with APL who were undergoing ATRA therapy (45 mg/m2 per day p.o.). Two of the patients were female, in whom this condition is quite rare. Genital ulcers with concomitant fever appeared between 17 and 32 days of therapy in all four patients. Genital ulcers healed in three of the patients while another patient developed Fournier's gangrene and underwent left testectomy. Ulcer healing was brought by either local or intravenous corticosteroids. Intravenous dexamethasone actually enabled continued ATRA administration in one patient, while ATRA was discontinued in other two patients. If corticosteroids cannot control progression of genital ulcers nor concomitant fever, ATRA administration should be discontinued so as not to induce Fournier's gangrene nor retionic acid syndrome. Our experience indicates the importance of recognizing genital ulcers associated with ATRA in order that appropriate countermeasures can be taken. PMID- 14738158 TI - PET scan evaluation of thymic mass after autologous peripheral blood stem-cell transplantation in an adult with non-Hodgkin's lymphoma. AB - We report the case of a 31-year-old man with anaplastic large-cell lymphoma successfully treated with chemotherapy who showed mediastinal widening 5 months after autologous stem-cell transplantation. CT scan and PET evaluations were consistent with the diagnosis of benign thymic hyperplasia. Because of the rapid and aggressive course of this type of lymphoma, and the progressive widening of the mass at CT scan, we performed a mediastinal biopsy that confirmed these findings, showing normal thymic tissue. This is the first case of benign thymic hyperplasia defined with FDG-PET and confirmed by histologic evaluation. PMID- 14738159 TI - Human herpes virus-8-negative primary effusion lymphoma in a patient with common variable immunodeficiency. AB - Primary effusion lymphoma (PEL) is a newly described high-grade B cell lymphoma developing in association with human herpes virus type 8 (HHV-8) in human immunodeficiency virus (HIV)-infecting individuals. Common variable immunodeficiency (CVID) is a primary immunodeficiency disease characterized by reduced serum immunoglobulin and heterogeneous clinical features. The risk of cancer in CVID patients is increased. Here, we describe a PEL that developed in the pleural and pericardial cavities of an HIV-negative and HHV-8-negative patient with CVID. PMID- 14738160 TI - Sense of humor, childhood cancer stressors, and outcomes of psychosocial adjustment, immune function, and infection. AB - The diagnosis, treatment, and side effects of childhood cancer have been described as extremely stressful experiences in the life of a child. Anecdotally, children report that a sense of humor helps them cope with the daily experiences of living with cancer; however, no research has examined sense of humor and childhood cancer stressors. This study investigated the effect of sense of humor on the relationship between cancer stressors and children's psychosocial adjustment to cancer, immune function, and infection using Lazarus and Folkman's theory of stress, appraisal, and coping. A direct relationship was observed between sense of humor and psychosocial adjustment to cancer, such that children with a high sense of humor had greater psychological adjustment, regardless of the amount of cancer stressors. A moderating effect was observed for incidence of infection. As childhood cancer stressors increase, children with high coping humor scores reported fewer incidences of infection than low scorers. PMID- 14738161 TI - Protective isolation in hemopoietic stem cell transplants: a review of the literature and single institution experience. AB - Princess Margaret Hospital for Children, Perth, Western Australia, is a pediatric bone marrow transplant center. This center has both laminar flow and HEPA- (high efficiency particulate air-)filtered rooms for children undergoing allogeneic and autologous transplantation. HEPA-filtered rooms on negative pressure are used to nurse oncology children with infectious diseases. Over the winter months of 2001, there was an increased demand for single rooms for children with infectious diseases. Over the same period, a number of transplants were planned. Consequently, to guide practice decisions, a review of the literature and a survey of nursing practice in Australian and North American pediatric oncology units were undertaken. Findings showed that protective isolation measures such as positive-pressure single rooms, low microbial diets, and strict hand washing should be used only for children requiring allogeneic transplants. Use of other isolation measures were found to be of no added value for transplantation. As autologous transplants are increasingly performed in outpatient clinics, these children should not require the same level of protective isolation. PMID- 14738162 TI - Parental involvement and family-centered care in the diagnostic and treatment phases of childhood cancer: results from a qualitative study. AB - Few research studies have addressed the ways parents participate in their childs medical care, particularly in relation to the cancer experience. The purpose of this study was to explore parents descriptions of their participation in medical care for their children with cancer. For this study, seven focus groups were conducted with 45 parents of 26 children who had completed cancer treatment at least one year prior, and who were still alive. Data were coded using thematic analysis procedures. It was found that parents emphasized their role as advocates during the diagnosis and treatment phases, by informing themselves about their childrens medical conditions, making medical care decisions, limiting the actions of medical professionals, and affirming and supporting medical professionals. These results emphasize the need to employ a family-centered approach in cancer care medical settings, by fostering and supporting the active inclusion of parents in their child's treatment and management. PMID- 14738163 TI - Superfecundation and superfetation: lessons from the past on early human development. PMID- 14738164 TI - East and West: from opposition to integration. PMID- 14738165 TI - Routine hemoglobin testing following an elective Cesarean section: is it necessary? AB - OBJECTIVE: To determine the clinical relevance of routine hemoglobin testing following an elective Cesarean section. METHODS: The charts of all women who underwent elective Cesarean section at a public teaching hospital during 2001 were reviewed for pre- and postoperative hemoglobin values, demographic data, indication for surgery and need for blood transfusion. RESULTS: The study group included 383 patients. Mean (+/- SD) hemoglobin level was 12.24 +/- 1.09 g/dl prior to surgery and 10.87 +/- 1.2 g/dl after, a mean loss of 1.37 +/- 0.87 g/dl (p < 0.001). There was no significant difference in blood loss by indication for surgery. None of the patients needed an intraoperative or postoperative blood transfusion. CONCLUSION: Routine postoperative hemoglobin measurement after an uncomplicated elective Cesarean section in asymptomatic low-risk women is not necessary and should be eliminated. PMID- 14738166 TI - Misoprostol administration in first-trimester pregnancies with embryonic demise reduces uterine arterial blood flow. AB - OBJECTIVES: This study was designed to evaluate the effect of misoprostol on the blood flow in uterine arteries of pregnant women with first-trimester embryonic demise. METHODS: Transvaginal ultrasonographic Doppler examinations (resistance index, pulsatility index, systolic/diastolic ratio) of the uterine arteries in 61 pregnant women were performed before misoprostol administration and 90 min later. Following baseline Doppler measurements, each woman received 200 microg misoprostol intravaginally and 200 microg misoprostol orally. RESULTS AND CONCLUSION: All Doppler indices increased significantly after misoprostol administration (p < 0.0001), suggesting an increase in flow resistance. PMID- 14738167 TI - Oral misoprostol is rapidly absorbed in postpartum women at term. AB - OBJECTIVE: To determine the rate of bioavailability of oral misoprostol in the tablet and a new capsule form in women with term pregnancies in the postpartum period. METHODS: Twenty-seven women received 400 microg of misoprostol orally after delivery of the fetal vertex in either the standard tablet form or crushed in methylcellulose capsules prepared in our pharmacy. Serum levels of misoprostol free acid, the principal metabolite, were measured at 5-, 15- and 30-min intervals after administration of the medication. The pharmacokinetics of the tablet and capsule groups were then compared. RESULTS: Twenty patients were included in the analysis. At 5 min, there was a trend towards a statistically significant difference in the concentration of misoprostol acid in the tablet group (89 pg/ml) versus the capsule group (20 pg/ml) (p = 0.007). No significant difference in plasma concentration was noted in the two groups at 15 min (tablet group, 256 pg/ml; capsule group, 245 pg/ml; p = 0.85) or 30 min (tablet group, 381 pg/ml; capsule group, 455 pg/ml; p = 0.45). CONCLUSION: Oral misoprostol is rapidly absorbed and bioavailable in the postpartum period. Misoprostol may prove useful in postpartum management. The novel packaging of misoprostol in capsule form allows for double-blinded studies with similar pharmacokinetics to the standard tablet. PMID- 14738168 TI - Comparing ambulatory spot urine protein/creatinine ratios and 24-h urine protein measurements in normal pregnancies. AB - OBJECTIVE: To determine whether a single specimen protein/creatinine ratio correlated with a 24-h urine protein measurement in ambulatory women with normal pregnancies. METHODS: A prospective cohort was evaluated of healthy women performing a 24-h urine collection and spot urine measurement for protein and creatinine in each trimester and the postpartum period. Data were analyzed to establish the correlation of values and the reliability of cut-off values. RESULTS: Fifty-eight women were enrolled in the study. Thirty-three completed at least one measurement. Three (5.1%) developed pre-eclampsia. With the use of linear regression, the spot protein/creatinine ratio correlated with the 24-h protein measurement in the first trimester and postpartum periods only (p < 0.001 and p < 0.043, respectively). It did not correlate in the second or third trimester (p = 0.637 and p = 0.290, respectively). CONCLUSION: In this population, the protein/creatinine ratio correlated only with the 24-h urine protein measurement in the first trimester and postpartum periods. In the periods of pregnancy where physiological changes are most prevalent, the ratio does not predict total urinary protein reliably. PMID- 14738169 TI - Comparison of obstetric outcomes in twin pregnancies after in vitro fertilization, ovarian stimulation and spontaneous conception. AB - OBJECTIVE: To compare the outcomes of liveborn twin gestations conceived after in vitro fertilization (IVF) or ovarian stimulation with spontaneously conceived twin pregnancies. METHODS: A review of all twin gestations delivered at Winthrop University Hospital from 1 January 1999 to 31 December 2000. Women who underwent fetal reduction or had a demise of one twin were excluded. Maternal demographics, antepartum complications, mode of delivery and perinatal outcome were compared. RESULTS: Sixty pregnancies were conceived after IVF, 34 were conceived by ovarian stimulation and 101 were spontaneously conceived. Women in the IVF group were older (p < 0.001), were more often 35 years or older (p < 0.001) and primiparous (p = 0.005). More women in the ovarian stimulation group had a poor obstetric history (p = 0.04). Spontaneous gestations had a higher incidence of monochorionic placentations (p = 0.002). There were no differences in gestational age at delivery, antepartum complications, or mode of delivery. There were fewer low-birth-weight neonates in the IVF group (odds ratio 0.59, 95% confidence interval 0.35-0.98; p = 0.03) than in the spontaneous group, but the difference disappeared when only the dichorionic pregnancies were compared. Other neonatal outcomes studied were the same among groups. CONCLUSION: Twin gestations conceived following IVF and ovarian stimulation appear to have similar outcomes to spontaneously conceived twin gestations. PMID- 14738170 TI - Unexplained fetal death is associated with changes in the adaptive limb of the maternal immune response consistent with prior antigenic exposure. AB - OBJECTIVE: The causes of fetal death are largely unknown. CD4 T cells have been classified according to the expression of the CD45 isoforms into 'naive-like' T cells (CD45RA) and 'memory-like' T cells (CD45RO). An increase in the percentage of the CD45RO has been interpreted as indicating prior antigenic exposure of the host and, in newborns, evidence of infection. The purpose of this study was to determine whether unexplained fetal death was associated with a change in the proportion of 'naive-like' and 'memory-like T cells' in the maternal blood, as determined by the CD45 isoforms on the surface of CD4+ lymphocytes. STUDY DESIGN: A prospective study was conducted to compare the CD45 sub-population of lymphocytes in patients with intrauterine fetal death (n = 26) and normal pregnancy (n = 89). The percentages of CD45RA+, CD45RO+ and CD45RA+/CD45RO+ on CD4+ T lymphocytes were determined in maternal blood using flow cytometry and monoclonal antibodies. Results were reported as a percentage of CD4+ lymphocytes. Non-parametric statistics were used for analysis. A p value of < 0.05 was considered significant. RESULTS: Patients with intrauterine fetal death had a higher percentage of CD45RO+ CD4+ T lymphocytes than normal pregnant women (fetal death: median 57.7%, range 35.4-78.6 vs. normal pregnancy: median 49.9%, range 19.1-86.8; p = 0.004). Fetal death was associated with a lower median percentage of CD45RA+ CD4+ lymphocytes than in normal pregnant women (fetal death: median 32.3%, range 15.3-58.0 vs. normal pregnancy: median 40.2%, range 11.2-67.3; p = 0.01). There was no significant difference in the percentage of cells with dual expression (CD45RA+/CD45RO+) between the study groups. CONCLUSION: Prior exposure to microbial products (bacterial or viral) or other unidentified antigens may result in a shift of the sub-population of 'naive-like' T cells to 'memory-like' T cells in mothers with unexplained fetal death. PMID- 14738171 TI - In vivo endoscopic assessment of arterioarterial anastomoses: insight into their hemodynamic function. AB - OBJECTIVE: To assess endoscopically the hemodynamic function of arterioarterial (AA) anastomoses in twin-twin transfusion syndrome (TTTS) and monochorionic selective intrauterine growth restriction (IUGR). MATERIALS AND METHODS: The videotapes of TTTS and IUGR patients undergoing laser surgery between July 1997 and December 2001 were reviewed for the presence of AA anastomoses. The hemodynamic equator was defined as the site within the AA anastomosis with color flashing. AA anastomoses were classified as having unidirectional flow, having bi directional flow, or being non-functional, depending on whether the hemodynamic equator reached a returning vein to one, both, or neither twin, respectively. TTTS was classified in stages as previously described. RESULTS: AA anastomoses were present in 35/183 (19.1%) of TTTS and in 12/24 (50%) IUGR patients. Of these, the hemodynamic equator was visible in 8/35 (22.8%) TTTS patients (all in stage III, and mostly in atypical stage III) and in 6/12 (50%) IUGR patients (overall 14/47, 29.8%). Of the 14 patients with a visible hemodynamic equator, 13 (92.8%) AA anastomoses showed unidirectional (9/13, 69.2% from the smaller to the larger twin) flow, and only 1/14 (7.1%) showed bi-directional flow. CONCLUSION: The hemodynamic equator is visible in approximately 30% of patients with AA anastomoses. Within this group, most AA anastomoses behave as functional arteriovenous anastomoses, and the direction of flow can be from the smaller to the larger twin or vice versa. The data suggest a correlation between sonographic findings and placental vascular design, also implying possible interfetal oxygenation differences. Further assessment of the functional behavior of AA anastomoses is warranted to understand the pathophysiology of TTTS and selective IUGR. PMID- 14738172 TI - Continuous glucose monitoring for the evaluation and improved control of gestational diabetes mellitus. AB - OBJECTIVES: To compare the daily glycemic profile reflected by continuous versus self-monitoring of blood glucose in women with gestational diabetes mellitus (GDM), and to evaluate possible differences in treatment strategy based on the two monitoring methods. MATERIALS AND METHODS: The study sample consisted of 57 women with gestational diabetes, 47 in Israel and ten in California. Gestational age ranged from 24 to 32 weeks in the Israeli women, and 32 to 36 weeks in the American women. Data derived from the Continuous Glucose Monitoring (CGM) System (MiniMed) for 72 h were compared to fingerstick glucose measurements (6-8 times a day). During continuous monitoring, patients documented the timing of food intake, insulin injections and hypoglycemic events. RESULTS: In the Israeli group, 23 women were treated by diet alone, and 24 by diet plus insulin. An average of 763 +/- 62 glucose measurements was recorded for each patient with continuous glucose monitoring. The mean total time of hyperglycemia (glucose level > 140 mg/dl) undetected by the fingerstick method was 132 +/- 31 min/day in the insulin-treated group and 94 +/- 23 min/day in the diet-treated group. Nocturnal hypoglycemic events (glucose levels < 50 mg/dl) were recorded in 14 patients, all insulin-treated. On the basis of the additional information provided by continuous monitoring, the therapeutic regimen (insulin therapy, diet adjustment, or both) was changed in 36 of the 47 patients. All ten American women were treated with insulin. The mean time of undetected hyperglycemia for a total group monitoring time of 30 days was 78 +/- 13 min/day. Eight women had nocturnal hypoglycemia on at least one of the three nights of monitoring for a total of 12 nights. A change in insulin dosage was made in all women on the basis of the data provided by continuous glucose monitoring. CONCLUSION: Continuous glucose monitoring is helpful for monitoring women with GDM and for adjusting diabetes therapy. It can accurately detect high postprandial blood glucose levels and nocturnal hypoglycemic events that may go unrecognized by intermittent blood glucose monitoring. A large prospective study on maternal and neonatal outcome is needed to determine the clinical implications of this new monitoring technique. PMID- 14738173 TI - Cocaine affects prostaglandin production in human umbilical cord cell cultures. AB - OBJECTIVE: To investigate the possible effects of cocaine on prostacyclin and prostaglandin (PG) E2 production from endothelial cells derived from human umbilical cord. STUDY DESIGN: First-passaged endothelial cells derived from the umbilical vein were incubated with various doses of cocaine, procaine and lidocaine and 24 h later the supematants were assayed for prostacyclin metabolites 6-keto-PGF1alpha and PGE2. Cocaine concentrations tested were 0, 10, 100, 500 and 1000 microg/ml. RESULTS: Cocaine produced a dose-dependent reduction in prostacyclin and PGE2 production from endothelial cells (p) < 0.05). Acetylcholinesterase (a possible detoxifier of cocaine) abolished the effect of cocaine on prostacyclin production. Procaine, an esterol-type anesthetic, produced a similar effect on prostacyclin production, an effect not observed with lidocaine. CONCLUSION: It is speculated that, when present in high concentrations, cocaine may affect vascular tone by inhibition of endothelial cell prostacyclin and PGE2 release. PMID- 14738175 TI - Umbilical cord transection in twin-reverse arterial perfusion syndrome with the use of a coaxial bipolar electrode (Versapoint). AB - Umbilical-cord occlusion has been proposed for the treatment of twin-reverse arterial perfusion syndrome (TRAP). Transection of the umbilical cord is necessary in patients with monoamniotic or 'pseudomonoamniotic' TRAP to avoid subsequent cord entanglement and demise of the pump twin. We present a case of TRAP in which the umbilical cord was successfully ligated with suture and transected with the Versapoint electrode. PMID- 14738174 TI - Suboptimal care and perinatal mortality in ten European regions: methodology and evaluation of an international audit. AB - BACKGROUND: A European concerted action (the EuroNatal study) investigated differences in perinatal mortality between countries of Europe. This report describes the methods used in the EuroNatal international audit and discusses the validity of the results. METHODS: Perinatal deaths between 1993 and 1998 in regions of ten European countries were identified. The categories of death chosen for the study were singleton fetal deaths at 28 or more weeks of gestational age, all intrapartum deaths at 28 or more weeks of gestational age and neonatal deaths at 34 or more weeks of gestational age. Deaths with major congenital anomalies were excluded. An international audit panel used explicit criteria to review all cases, which were blinded for region. Subjective interpretation was used in cases of events or interventions where explicit criteria did not exist. Suboptimal factors were identified in the antenatal, intrapartum and neonatal periods, and classified as 'maternal/social', due to 'infrastructure/service organization', or due to 'professional care delivery'. The contribution of each suboptimal factor to the fatal outcome was listed and consensus was reached on a final grade using a procedure that included correspondence and plenary meetings. RESULTS: In all regions combined, 90% of all known or estimated cases in the selected categories were included in the audit. In total, 1619 cases of perinatal death were audited. Consensus was reached in 1543 (95%) cases. In 75% of all cases, the grade was based on explicit criteria. In the remaining cases, consensus was reached within subpanels without reference to predefined criteria. There was reasonable to good agreement between and within subpanels, and within panel members. CONCLUSIONS: The international audit procedure proved feasible and led to consistent results. The results that relate to suboptimal care will need to be studied in depth in order to reach conclusions about their implications for assessing the quality of perinatal care in the individual regions. PMID- 14738176 TI - Twin-twin transfusion syndrome in a dizygotic monochorionic-diamniotic twin pregnancy. AB - We present a case of twin-twin transfusion syndrome with discordant gender. Monochorionicity was confirmed by surgical pathology. Cytogenetic analysis showed normal 46,XX and 46,XY karyotypes. Microsatellite analysis using reliable pericentromeric markers was consistent with dispermic fertilization of two separate ova. This suggests that monochorionicity, rather than zygosity, may be responsible for the development of placental vascular anastomoses. PMID- 14738177 TI - A chart review study of the inattentive and combined types of ADHD. AB - Studies of the clinical correlates of the subtypes of Attention Deficit/Hyperactivity Disorder (ADHD) have identified differences in the representation of age, gender, prevalence, comorbidity, and treatment. We report retrospective chart review data detailing the clinical characteristics of the Inattentive (IA) and Combined (C) subtypes of ADHD in 143 cases of ADHD-IA and 133 cases of ADHD-C. The children with ADHD-IA were older, more likely to be female, and had more comorbid internalizing disorders and learning disabilities. Individuals in the ADHD-IA group were two to five times as likely to have a referral for speech and language problems. The children with ADHD-IA were rated as having less overall functional impairment, but did have difficulty with academic achievement. Children with ADHD-IA were less likely to be treated with stimulants. One eighth of the children with ADHD-IA still had significant symptoms of hyperactivity/impulsivity, but did not meet the DSM-IV threshold for diagnosis of ADHD-Combined Type. The ADHD-IA subtype includes children with no hyperactivity and children who still manifest clinically significant hyperactive symptomatology but do not meet DSM-IV criteria for Combined Type. ADHD-IA children are often seen as having speech and language problems, and are less likely to receive medication treatment, but respond to medical treatment with improvement both in attention and residual hyperactive/impulsive symptoms. PMID- 14738178 TI - Diagnostic utility of DSM-IV-TR symptoms in the prediction of DSM-IV-TR ADHD subtypes and ODD. AB - OBJECTIVE: Examined the diagnostic utility of DSM-IV-TR symptoms in the prediction of DSM-IV-TR ADHD subtypes and ODD to: (a) Assess the relative utility of parent and teacher ratings, and (b) Examine the utility of individual symptoms as inclusion and exclusion criteria. METHOD: Parent and teacher versions of the Disruptive Behavior Disorders Rating Scale were completed for 440 children (217 boys; 223 girls) ranging in age from 9 to 13 years and were used to classify children into groups. Individual symptoms were used to compute corrected conditional probability statistics. RESULTS: Corrected positive predictive power (cPPP) statistics indicated that parent and teacher ratings had similar diagnostic utility in the prediction of the presence of ADHD, Inattentive Type and ADHD, Hyperactive/Impulsive Type, and that their combination did not provide incremental utility over either informant alone. However, cPPP statistics for ADHD, Combined Type, and ODD indicated that teacher ratings, and the use of combined parent and teacher ratings, provided an advantage over parent ratings alone. Results support the use of a multi-method, multi-informant assessment approach. PMID- 14738179 TI - Parenting of children with attention-defecit/hyperactivity disorder (ADHD): the role of parental ADHD symptomatology. AB - OBJECTIVE: The present study examined the relation between parental ADHD symptomatology and parent-child behavior among 46 mothers and 26 fathers of ADHD children. RESULTS: Fathers' self-reports of inattention and impulsivity were strongly associated with self-reports of lax parenting both before and after parent training, and with self-reports of overreactivity after parent training. Fathers' impulsivity was also associated with more arguing during audiotaped observations of parent-child interactions prior to parent training. Mothers' self reports of inattention were modestly associated with self-reports of laxness before and after parent training. Prior to parent training, there were non-linear relations between mothers' inattention and observations of mother-child behavior, with mothers who reported moderate levels of inattention engaging in the most negative parent-child interactions. After parent training, these relations were linear, with the mothers who reported the most inattention engaging in the most negative parent-child interactions. These results were weakened but were generally still significant when parental depression and alcohol use were controlled. PMID- 14738180 TI - EEG biofeedback vs. placebo treatment for attention-deficit/hyperactivity disorder: a pilot study. AB - METHOD: Seven children diagnosed with attention-deficit hyperactivity disorder (ADHD) were trained using a standard EEG biofeedback treatment protocol designed to alter SMR/theta ratios and reduce behavioral symptomatology diagnostic of ADHD. During alternate periods they were also trained using a placebo protocol that was identical to the treatment protocol, save that the association between EEG patterns and feedback to the participants was random. Single-case design elements were used to control for the effects of internal validity threats such as maturation, history, and treatment order. Two participants failed to complete all training sessions, and the effects of training on behavior were analyzed both including and excluding these non-completers. RESULTS: When all participants were included in analyses that controlled for overall trend, EEG biofeedback was found to be no more effective than the placebo control condition involving non contingent feedback, and neither procedure resulted in improvements relative to baseline levels. When overall behavioral trends unrelated to training were not controlled for and non-completers were excluded from the analysis, it could be mistakenly concluded that EEG biofeedback is significantly more effective than placebo and that the effect sizes involved are moderate to large. These results indicate that many previous reports of the efficacy of EEG biofeedback for ADHD, particularly those presenting series of single cases, might well have been based on spurious findings. PMID- 14738181 TI - Integration of clinical practice, publicity, and policy: a shot in the arm for influenza control. PMID- 14738182 TI - The physician's office: can it influence adult immunization rates? AB - OBJECTIVE: To determine which office and patient factors affect adult influenza and pneumococcal vaccination rates. STUDY DESIGN: Patient interviews and self administered surveys of office managers. PATIENTS AND METHODS: In a 2-stage random cluster sample, 22 practices in 4 strata (Veterans' Affairs, rural, urban/suburban, and inner city) and 15 patients per physician in each practice (n = 946) were selected. Office managers completed a questionnaire regarding office practices and logistics affecting immunizations. Data were examined using chi2 and regression analyses without and with patient factors in the models. RESULTS: Practice factors significantly related to influenza vaccination status were stratum (VA OR = 2.04; 95% CI = 1.18, 3.53; P < .05 vs inner-city), time allotted for acute care visits (16-20 min vs 10-15 min OR = 2.49; 95% CI = 1.68, 3.09; P < .001), the practice not having a source of free vaccines (OR = .43; 95% CI = .3, .62; P < .001), and the interaction between being an urban/suburban practice and having a source of free flu vaccines (OR = 4.0; 95% CI = 2.63, 6.09; P < .001). Practice factors related to pneumococcal vaccination status were the number of immunization promotion activities (> or = 3 vs 0-2 OR = 1.97; 95% CI = 1.33, 2.94; P = .002) and the time allotted for acute care visits (16-20 min vs 10-15 min OR = 1.94; 95% CI = 1.18, 3.19; P = .011). When practice and patient factors were combined in the analyses, patient factors were more important. CONCLUSION: Although patient factors are more important than practice factors, practices that allot more time for acute care visits and use more immunization promotion activities have higher vaccination rates. PMID- 14738183 TI - The rate, pattern, and cost of use of antiparkinsonian agents among patients treated for schizophrenia in a managed care setting. AB - OBJECTIVE: To examine the use and economic cost of antiparkinsonian agents during the medication management of patients diagnosed with schizophrenic disorders in a naturalistic healthcare setting. DESIGN: Cross-sectional retrospective analysis of 1-year (1999) administrative data from a large managed care organization. PATIENTS AND METHODS: Patients were 1938 adults who were treated for a schizophrenic disorder. Monthly per patient utilization rate and cost of antiparkinsonian agents, and the proportion of the total psychiatric medication costs attributed to the antiparkinsonian agents, were compared across antipsychotic medications (typical, atypical, olanzapine, risperidone), age, sex, and diagnostic subtype. RESULTS: About one third (39%) of the patients were treated with typical antipsychotics only, and half of them (51%) received antiparkinsonian agents. Those treated with atypical antipsychotics only (41%) were considerably less likely to receive antiparkinsonian agents (25%), and the rate of use differed by atypical type such that risperidone-treated patients were more likely to receive antiparkinsonian agents than those treated with olanzapine. Lower utilization rates of antiparkinsonian agents were also found among patients age 75 years or older, and among those diagnosed with a schizoaffective disorder. Average monthly per patient cost of antiparkinsonian agents was 3.0 dollars, constituting 2.6% of the monthly expenditure on all psychiatric medications. CONCLUSIONS: Adjunctive use of antiparkinsonian agents differs widely among patients who are treated with typical or atypical antipsychotic drugs, and differs between types of atypical antipsychotics. The choice of atypical antipsychotics that have a lower liability for extrapyramidal symptoms may assist in optimizing the long-term functional outcomes of schizophrenia patients. PMID- 14738184 TI - Asthma population management: development and validation of a practical 3-level risk stratification scheme. AB - OBJECTIVE: To define and validate a practical risk stratification scheme based on administrative data for use in identifying patients at high, medium, and low risk of requiring emergency hospital care for asthma. STUDY DESIGN: Retrospective cohort. PATIENTS AND METHODS: Predictors in 1999 were evaluated in relation to 2000 asthma emergency hospital care (any asthma hospitalization or emergency department visit) in a training set (n = 8789, 2000 emergency hospital care = 5.5%) and a testing set (n = 6104, 2000 emergency hospital care = 7.9%). Logistic regression was used to assign risk points in the training set, and positive and negative predictive values, sensitivities, and specificities were calculated in the training and testing sets. RESULTS: High risk was defined as asthma emergency hospital care in the previous year or use of >14 beta-agonist canisters and oral corticosteroid use; medium risk was defined as no emergency hospital care but use of either >14 beta-agonist canisters or oral corticosteroids; and low risk was defined as none of the above. For the high-risk groups in the training and testing sets, positive predictive values were 12.9% and 22.0%, sensitivities were 24.8% and 25.4%, specificities were 90.3% and 92.0%, and negative predictive values were 95.4% and 93.2%, respectively. The medium-risk groups identified another 32.6% of patients in the training set and 28.3% in the testing set requiring subsequent asthma emergency hospital care. CONCLUSION: This simple risk stratification scheme is useful for identifying patients from administrative data who are at increased risk of experiencing emergency hospital care for asthma. PMID- 14738185 TI - Determinants of hospital length of stay for cervical dysplasia and cervical cancer: does managed care matter? AB - OBJECTIVE: To examine whether type of health insurance plan, among other variables, affects the length of stay for cervical cancer-related hospitalizations. STUDY DESIGN, PATIENTS, AND METHODS: Inpatient admission claims records for cervical dysplasia and cervical cancer were selected for 1994-1997 from the MarketScan private health insurance claims database. After identifying records by stage of disease and deleting records for pregnant women, 1145 unique patient records were used in a truncated count regression model to analyze the predictors of hospital length of stay. RESULTS: All later stages of disease were associated with a longer hospital stay. After controlling for other variables, the coefficients showed an increase in predicted length of admission ranging from 2.5 days for stage I to 6.3 days for stage IV cervical cancer compared with dysplasia/carcinoma in situ (all stages, P < .01). There was no significant statistical difference in the lengths of stay for patients covered under comprehensive fee-for-service plans vs other types of health insurance plans, including managed care. CONCLUSIONS: Managed care plans are often thought to contain healthcare costs by shortening the hospital length of stay. Our findings show no association between managed care plans and hospital length of stay for women with cervical cancer or its precursors. PMID- 14738186 TI - Employee responses to health insurance premium increases. AB - OBJECTIVE: To determine the sensitivity of employees' health insurance decisions- including the decision to not choose health maintenance organization or fee-for service coverage--during periods of rapidly escalating healthcare costs. STUDY DESIGN: A retrospective cohort study of employee plan choices at a single large firm with a "cafeteria-style" benefits plan wherein employees paid all the additional cost of purchasing more generous insurance. METHODS: We modeled the probability that an employee would drop coverage or switch plans in response to employee premium increases using data from a single large US company with employees across 47 states during the 3-year period of 1989 through 1991, a time of large premium increases within and across plans. RESULTS: Premium increases induced substantial plan switching. Single employees were more likely to respond to premium increases by dropping coverage, whereas families tended to switch to another plan. Premium increases of 10% induced 7% of single employees to drop or severely cut back on coverage; 13% to switch to another plan; and 80% to remain in their existing plan. Similar figures for those with family coverage were 11%, 12%, and 77%, respectively. Simulation results that control for known covariates show similar increases. When faced with a dramatic increase in premiums--on the order of 20%--nearly one fifth of the single employees dropped coverage compared with 10% of those with family coverage. CONCLUSIONS: Employee coverage decisions are sensitive to rapidly increasing premiums, and single employees may be likely to drop coverage. This finding suggests that sustained premium increases could induce substantial increases in the number of uninsured individuals. PMID- 14738187 TI - Ionic dialysance. PMID- 14738188 TI - Influence of hepatitis B virus virema upon serum aminotransferase activity in dialysis population. AB - BACKGROUND: The control of the spread of hepatitis B virus (HBV) infection within dialysis units has been one of the major advances in the management of patients with end-stage renal disease (ESRD). However, clinical and biochemical expression of HBV in dialysis patients have not been adequately addressed. Elevated values of serum aminotransferase activity are a sensitive measure of hepatocellular injury, but the role of HBV infection in the development of liver disease among dialysis patients has not been adequately analysed. Also, the clinical impact related to the virological characteristics of HBV in dialysis has not been evaluated. METHODS: Demographic, biochemical and virological data from 727 patients undergoing chronic dialysis in seven dialysis units in northern Italy were collected in order to assess the biochemical consequences related to the presence of HBV infection in this population. We have measured by RT-PCR technology the titers of HBV viremia in HBsAg positive patients receiving dialysis. RESULTS: Univariate analysis showed that AST and ALT values were significantly higher in HBsAg positive/HBV DNA positive than HBsAg negative patients on dialysis; AST, 22.86+/-31.34 vs. 14.19+/-9.7 IU/L (P=0.00001); and ALT, 25.07+/-41.59 vs. 13.9+/-41.59 IU/L (P=0.00001). In the subgroup of HBsAg positive patients, the frequency of detectable HBeAg in serum was 14.9% (7/47). The median value of HBV DNA in patients with detectable HBV DNA in serum was 2.160 x 10(3) copies/mL (range, 2.5 x 10(2)-4 x 10(6) copies/mL). HBsAg positive/HCV positive patients had higher aminotransferase activity than other subgroups (P=0.0001). Multivariate analysis showed a significant and independent association between detectable HBsAg/HBV DNA in serum and AST (P=0.00001) and ALT (P=0.0001) activity AST and ALT levels were lower in dialysis than healthy individuals--this finding persisted in age- and gender-matched comparisons. CONCLUSIONS: The HBV viral load in HBsAg positive patients receiving maintenance dialysis is not high. HBsAg positivity with detectable HBV DNA in serum is a strong and independent predictor of raised aminotransferase activity among dialysis patients. HBsAg positive patients had greater aminotransferase activity than HBsAg negative individuals even if both the groups had mean aminotransferase levels within the normal range considered for healthy population. Clinical trials aimed at identifying the best cut-off value to enhance the diagnostic yield of AST/ALT for detecting HBV in dialysis population are under way. PMID- 14738189 TI - Comparison of different techniques of hemodialysis vascular access flow evaluation. AB - Measurement of vascular access flow (QVA) has been suggested as a method of choice for vascular access quality (VAQ) monitoring. Besides traditional duplex Doppler, a number of bedside methods based mostly on the Krivitski principle of QVA evaluation from recirculation at reversed needles (RX), have been developed. This work compares ultrasonic dilution (UD), taken as a reference, HD01, Transonic Systems; duplex Doppler (DD); thermodilution (TD), BTM, Fresenius; optodilutional RX measurement (ORX), Critline III, R-mode, HemaMetrics; direct optodilutional QVA evaluation from jumpwise changes in ultrafiltration rate at both normal and reversed needles connection (OABF), Critline III, ABF-mode; and direct transcutaneous optodilutional QVA evaluation (TQA), Critline III TQA. Firstly, reproducibility of each method was assessed by duplicate measurement at unchanged conditions. This was followed by paired measurement with each method performed at controlled change in relevant measurement condition (two different extracorporeal blood flows in UD and TD, changed sensor position in TQA). Finally paired measurements by each method and the reference method performed at identical conditions were evaluated to assess accuracy of each method. The simple Krivitski formula QVA=QB(1-RX)/RX was used wherever manual QVA calculation was needed. Very high reproducibility was seen in UD, both for measurement at the same extra corporeal blood flow (QB) (correlation coefficient of duplicate measurement r=0.9702, n=58) and for measurement at two different QB (r=0.9735, n=24), justifying its current status of a reference method in QVA evaluation. Slightly lower reproducibility of TD measurement at the same QB (r=0.9197, n=40) and at two different QB (r=0.8508, n=168) can be easily overcome by duplicate measurement with averaging. High correlation of TD vs. UD (r=0.9543, n=54) makes TD a viable clinical alternative in QVA evaluation. Consistently different QVA obtained at two different QB should prompt closer investigation of anatomical conditions of the access. Use of the simple Krivitski formula in TD (which measures total recirculation, i.e. sum of access recirculation and cardiopulmonary recirculation) brings about underestimation of QVA, which progressively increases from QVA of about 600 mL/min up. Good correlation, although with significant scatter (r=0.8691, n=27) was found between the DD- and UD-based QVA. By far the worst reproducibility at the same QB from among the investigated methods was found in ORX (0.6430, n 23). Also the correlation of ORX vs. UD was lower than in other methods (r=0.702, n=33) and general overestimation of QVA by about 25% was noted. Correlation of OABF vs. UD (r=0.6957, n=26) was slightly better than that of ORX and it gave less overestimated values. The TQA method showed very high reproducibility (r=0.9712, n=85), however only for unchanged sensor position. Correlation of QVA measured at two different sensor positions was much worse (r=0.7255, n=22). Correspondence of TQA vs. UD was satisfactory (r=0.8077, n=36). Skilled and experienced operators are a must with this method. PMID- 14738190 TI - Preadipocyte-loaded collagen scaffolds with enlarged pore size for improved soft tissue engineering. AB - Extended soft tissue defects after extensive deep burns or tumor resections are still an unresolved problem in plastic and reconstructive surgery. There is a clinical need for an adequate solution to this problem but currently, no adequate implant material is available for the correction of these defects. Since the autologous transplantation of mature adipose tissue gives poor results, this study explores the advantages of using human preadipocytes in collagen sponges for tissue reconstruction purposes. Human preadipocytes of young adults were isolated, cultured, seeded onto collagen sponges with uniform pore size, and implanted into immunodeficient mice. After 24 hours of incubation in vitro and after explantation at 3, 8, and 12 weeks, sponges were examined for macroscopic appearance, weight, thickness, histology, immuno-histochemistry, and ultrastructure. We find good penetration of cells into the scaffold, layers of adipose tissue, and new vessels on all grafts while controls appear unchanged. These results are promising for improving the reconstruction of soft tissue defects. PMID- 14738191 TI - Tailoring biomaterial compatibility: in vivo tissue response versus in vitro cell behavior. AB - Biocompatibility relies essentially on surface phenomena, represented by cell cell, cell-material and material (polymer)-protein interactions. An in vivo and in vitro experimental investigation was carried out on the biomaterials of two different classes with a good potential for in situ utilisation. Non-resorbable (Polypyrrole, Polyaniline, Polyimide) and resorbable (PLLA-PDXO-PLLA) materials for tissue engineering were studied for their overall tissue tolerance and cellular interactions. These non-resorbable polymers conceived for biosensor applications and implantable drug-delivery systems are intrinsically conductive. The PLLA-PDXO-PLLA triblock copolymer showed interesting tensile properties for bone and cartilage tissue engineering due to the presence of 1,5-dioxepan-2-one. In vitro and in vivo parallel studies showed an interesting correspondence: a) the cells in contact with the resorbable material that appeared to be capable of migratory-regenerative aspects in vitro exhibited good compatibility in vivo; whereas b) the non-resorbable materials, which are designed to remain in situ in vivo, were seen to have the potential to represent an adverse factor (inflammation, fibrotic reactions) that correlated with some aspects of cell behaviour in vitro. PMID- 14738192 TI - Platelet dysfunction in cardiopulmonary bypass: an experimental comparative study between a centrifugal and a new pulsatile pump. AB - The aim of this investigation was to study the effect of a new pulsatile pump for extracorporeal circulation (ECC) on platelet count and platelet function with respect to a Biomedicus centrifugal pump. Thirteen pigs, 8 in the pulsatile group (PG) and 5 in the centrifugal group (CG), underwent a partial extracorporeal circulation lasting 3 h. The animals were sacrificed 3 h post-ECC. The platelet study was both quantitative (platelet count) and qualitative (platelet function analysis) by assessing the closure time (CT) with a PFA-100 system. The decrease in platelet number from basal to 3 h post ECC was only significant in CG (p = 0.009). The platelet function was impaired in both groups, but the value of CT with col/ADP increased significantly only in CG (p < 0.001). The increase of CT with col/EPI was greater in CG (p = 0.07) than in PG (p = 0.2). The results indicated that the new pulsatile pump preserves platelets quantitatively and qualitatively well compared to a Biomedicus pump. PMID- 14738193 TI - The ex-vivo-shunt-model: novel approach for assessing the thrombogenicity of vascular implants. AB - OBJECTIVE: Disadvantages associated with commercially available vascular implants necessitate alternative strategies to develop new vascular prostheses. Although many tissue characterizing strategies have been defined, no valid test for thrombogenicity exists. Here we introduce a novel concept for thrombogenicity testing of vascular implants METHODS: Silastic tubes were implanted into the carotid arteries of 12 sheep. After placing these shunts, tc99m-labeled platelets were administered and test-vessels were put in between the shunts. Native autologous (n=6), as well as native/acellularized allogeneic (n=6/n=6), and xenogeneic (n=6/n=6) carotid arteries and allogeneic (n=6/n=6) and xenogeneic (n=6/n=6) carotid arteries reseeded with allogeneic endothelial-cells, fibroblasts and myocytes were evaluated. Number and time course of intra operatively deposited platelets were evaluated with a Geiger-counter; certain areas of platelet deposition located, envisioned and characterized by a gamma camera and scanning electron-microscopy afterwards. RESULTS: Counter results revealed no significant different platelet depositions when comparing silastic tubes with either autologous or allogeneic native carotid arteries. However, starting 5 minutes after placement, acellularized/reseeded allogeneic (p=0.001/p=0.00004), and xenogeneic (p=0.0001/p=0.01) carotid arteries showed significantly more platelet depositions than native autologous carotides. Moreover, it was possible to show that almost no platelets adhere to native vessels or silastic tubes, thus proving the test method itself. CONCLUSION: The Ex-Vivo-Shunt-Model is a valid method to measure and envision the intrinsic thrombogenicity of vascular implants. PMID- 14738194 TI - Modelling of cardiovascular system: development of a hybrid (numerical-physical) model. AB - Physical models of the circulation are used for research, training and for testing of implantable active and passive circulatory prosthetic and assistance devices. However, in comparison with numerical models, they are rigid and expensive. To overcome these limitations, we have developed a model of the circulation based on the merging of a lumped parameter physical model into a numerical one (producing therefore a hybrid). The physical model is limited to the barest essentials and, in this application, developed to test the principle, it is a windkessel representing the systemic arterial tree. The lumped parameters numerical model was developed in LabVIEW environment and represents pulmonary and systemic circulation (except the systemic arterial tree). Based on the equivalence between hydraulic and electrical circuits, this prototype was developed connecting the numerical model to an electrical circuit--the physical model. This specific solution is valid mainly educationally but permits the development of software and the verification of preliminary results without using cumbersome hydraulic circuits. The interfaces between numerical and electrical circuits are set up by a voltage controlled current generator and a voltage controlled voltage generator. The behavior of the model is analyzed based on the ventricular pressure-volume loops and on the time course of arterial and ventricular pressures and flow in different circulatory conditions. The model can represent hemodynamic relationships in different ventricular and circulatory conditions. PMID- 14738195 TI - Robin Heart 2003--present state of the Polish telemanipulator project for cardiac surgery assistance. AB - The Polish telemanipulator (Robin Heart), for use in cardiac surgery, has been realized by the Foundation of Cardiac Surgery Development in Zabrze, Poland, in cooperation with specialists from the Technical University of Lodz and Warsaw University of Technology. The brief history of robotic surgery and fundamental advantages of employing robots in this field--safe, reliable and repeatable operative results with less patient pain, trauma and recovery time--follow the assumptions of the Polish Cardio-Robot project. The cardiac surgery robot, Robin Heart, is an original construction with a segment type structure which allows the various combination of its parts for different types of surgery. The telemanipulator for cardiac surgery will consist of two arms equipped with tools and one arm holding the camera. Several models suitable for surgeon contact systems, using the experience of centers designing the artificial hand and haptic systems have been worked out. The detailed mechanical analysis and original construction of main parts of the robot and development of the surgical planning system are presented in further sections. PMID- 14738196 TI - Understanding the tropospheric transport and fate of agricultural pesticides. AB - Many field monitoring studies have indicated the substantial role of the troposphere as both a sink and transport medium for pesticides. At the same time, this is the least studied and understood environmental compartment in regards to pesticide fate. Although the fundamental principles behind volatilization and tropospheric reactivity are well understood, it is becoming increasingly apparent that the ability to quantitatively measure flux and reaction rates in the air will continue to pose problems for researchers. To date, most deterministic models that try to simulate real world conditions generally fail to provide tropospherically relevant flux and reaction rates because it is virtually impossible to scale down all possible interactions occurring in a near-infinite reservoir. Better field methods for determining flux are emerging and more ambient air monitoring studies are being conducted. This growing database of information, together with an understanding of physicochemical properties and use of expert systems, has increased the predictive capability for estimating volatilization flux of pesticides. Unfortunately, there are very limited environmental data on the tropospheric reaction rates of pesticides, and more experimental studies using semivolatile to low-volatility pesticides or their more volatile homologues are required to validate existing structure-activity relationship (SAR) model predictions. The development of new analytical strategies using elevated temperatures for assessing semivolatile to low volatility pesticide reaction rates and products may provide an alternative approach to the need for controlled environmental temperature data. Recent international workshops organized by the Health Council of The Netherlands for developing uniform approaches for assessing exposure risks to pesticides in air exemplify efforts to synchronize flux with environmental fate information for determining human health and ecological risks. When more detailed pesticide information is desired, especially in high-use agricultural areas, where exposure to humans and nontarget ecological communities is a major concern, field flux measurements and downwind monitoring, together with experimental fate studies, should be considered. The integration of models, empirical testing, and real world monitoring will provide the ultimate safety net needed for assessing exposure risks to airborne pesticides. PMID- 14738197 TI - Environmental lead in Mexico, 1990-2002. AB - From the data presented here, it can be concluded that despite some important progress in recent years, in particular, the phaseout of the use of lead in gas, environmental exposure to lead remains a particularly severe problem in Mexico. The lack of recent and adequate information on the presence of lead in air, dust, soil, surface waters, the marine environment, and food is noteworthy. In particular, there are no data on the presence of lead in the environment of industrial cities or in areas where mining and metallurgical activities are predominant, including their nearby environment, such as river sediments or air, and in their inhabitants. As a result, the official conclusion that the major present source of lead for Mexicans is the manufacture and use of lead-glazed ceramics, and the few official actions in this regard lack a solid basis because they rely on data from a few limited studies carried out mainly in Mexico City and its Metropolitan Zone. Although there have been few studies in Mexico on the adverse effects of long-term exposure to lead, the data provided on its presence in human milk, lungs, and bones and the results of the scant studies on its neuropsychological effects on children should be taken into consideration to establish more effective control and protective measures and to support the additional studies required to document the present situation. In particular, more studies should be carried out in lead glazed-ceramic-producing communities and on the hazards of lead to the communities established near industrial areas and along river banks. In future studies, special attention should be paid to establishing a common protocol, with adequate analytical control measures, if possible, with intercalibration of the participating laboratories; this is essential, because at present many of the available data cannot be compared or extrapolated due to their deficiencies in this regard. The situation around the only primary lead smelter in Mexico, Met Mex Penoles, is a relevant example. Although to date the government and the company have supported the determination of lead in more than 11,000 samples of children's blood, the analyses were carried out in several laboratories, without the minimum requirements to allow them to be valid or compared; thus, the number of children with lead above the different levels recommended by the CDC to establish their risk is not known. In closing, there is still much to be done to document the presence of lead in the environment in Mexico and its adverse effects for health and ecological equilibrium in order to control and reduce its sources; this will require a sustained effort from research institutions and continued support of the authorities, particularly those responsible for health and environment matters. Until these goals are achieved, it will not be possible to state that lead in the Mexican environment is no longer a problem or that it is restricted to particular sources or populations. It is also important to strengthen the legal framework of control to provide adequate enforcement and oversight measures; otherwise, both the existing standards and the new ones will be useless to improve the situation. PMID- 14738198 TI - Pesticides in Uruguay. AB - This is a review, from an independent and scientific point of view, of the different aspects involved in the issue of pesticides in Uruguay. In its preparation, the University, responsible official institutions, nongovenment environmental organizations, and independent experts have been consulted. As to the legal framework, the responsibilities of the institutions are described and evaluated. The earlier and current regulations for the registration and sale of pesticides are presented, as well as the available information on the active ingredients most used in Uruguay. Official control is evaluated in reference to pesticide residues in food, drinking water, and the environment and to final waste disposal. Maximum allowed residue limits and the responsibilities of the corresponding governmental institutions are presented. Emphasis is placed on the fact that there are no publicly available data. Several research programs are presented, mainly from academia, and usually are not completed due to lack of financial support. In the conclusions the most problematic aspects are pointed out, emphasizing the need to improve national regulations for the country to establish an effective system of control. The importance of financial support to achieve this control and to conduct interdisciplinary studies to determine the real situation is discussed. All compounds are listed in Table 19. PMID- 14738199 TI - Mercury hazards from gold mining to humans, plants, and animals. AB - Mercury contamination of the environment from historical and ongoing mining practices that rely on mercury amalgamation for gold extraction is widespread. Contamination was particularly severe in the immediate vicinity of gold extraction and refining operations; however, mercury, especially in the form of water-soluble methylmercury, may be transported to pristine areas by rainwater, water currents, deforestation, volatilization, and other vectors. Examples of gold mining-associated mercury pollution have been shown for Canada, the U.S., Africa, China, the Philippines, Siberia, and South America. In parts of Brazil, for example, mercury concentrations in all abiotic materials, plants, and animals, including endangered species of mammals and reptiles, collected near ongoing mercury amalgamation gold mining sites were far in excess of allowable mercury levels promulgated by regulatory agencies for the protection of human health and natural resources. Although health authorities in Brazil are unable to detect conclusive evidence of human mercury intoxication, the potential exists in the absence of mitigation for epidemic mercury poisoning of the mining population and environs. In the U.S., environmental mercury contamination is mostly from historical gold mining practices, and portions of Nevada remain sufficiently mercury contaminated to pose a hazard to reproduction of carnivorous fishes and fish-eating birds. Concentrations of total mercury lethal to sensitive representative natural resources range from 0.1 to 2.0 microg/L of medium for aquatic organisms; from 2,200 to 31,000 microg/kg BW (acute oral) and from 4,000 to 40,000 microg/kg (dietary) for birds; and from 100 to 500 microg/kg BW (daily dose) and from 1,000 to 5,000 microg/kg diet for mammals. Significant adverse sublethal effects were observed among selected aquatic species at water concentrations of 0.03-0.1 microg Hg/L. For some birds, adverse effects, mainly on reproduction, have been associated with total mercury concentrations (microg/kg FW) of 5,000 in feathers, 900 in eggs, and 50-100 in diet, and with daily intakes of 640 microg/kg BW. Sensitive nonhuman mammals showed significant adverse effects of mercury when daily intakes were 250 microg/kg BW, when dietary levels were 1,100 microg/kg, or when tissue concentrations exceeded 1,100 microg/kg. Proposed mercury criteria for protection of aquatic life range from 0.012 microg/L for freshwater life to 0.025 microg/L for marine life; for birds, less than 100 microg/kg diet FW; and for small mammals, less than 1,100 microg/kg FW diet. All these proposed criteria provide, at best, minimal protection. PMID- 14738200 TI - Laboratory assessment of gastrointestinal function. AB - Over the past decade, several laboratory tests have been introduced to veterinary medicine that allow the minimally invasive assessment of diseases of the gastrointestinal tract, liver, and pancreas. Although some of those tests have limited clinical use in a practice setting and have more use as research tools, other tests, such as serum cobalamin and folate concentrations, find wide application in everyday veterinary practice. In some cases, definitive diagnosis requires invasive or expensive procedures, but these new laboratory tests have greatly aided veterinarians in diagnosing gastrointestinal diseases. This discussion provides an overview of new diagnostic tests that will allow the minimally invasive assessment of gastrointestinal function. PMID- 14738201 TI - Nutritional management of gastrointestinal disease. AB - The gastrointestinal (GI) tract is primarily responsible for acquiring and digesting food, absorbing nutrients and water, and expelling wastes from the body as feces. A proper diet and normally functioning GI tract are integral for the delivery of nutrients, prevention of nutrient deficiencies and malnutrition, repair of damaged intestinal epithelium, restoration of normal luminal bacterial populations, promotion of normal GI motility, and maintenance of normal immune functions (eg, both tolerance and protection from pathogens). The amount of food, its form, the frequency of feeding, and the composition of diet each have important effects on GI function and may be used to help ameliorate signs of GI disease. Although both nutrients and nonnutritional components of a diet are important to GI health, they also may cause or influence the development of GI pathology (eg, antibiotic responsive diarrhea, inflammatory bowel disease, dietary intolerance, or sensitivity and/or allergy). The appropriate diet may have a profound effect on intestinal recovery and successful management of chronic or severe GI disease. PMID- 14738202 TI - Optimal fecal assessment. AB - Fecal testing is a common component of most gastrointestinal work-ups. A multitude of diagnostic techniques are available for identifying parasites and pathogens, or showing abnormalities of flora. Optimal fecal assessment involves careful formulation of a differential list based on signalment, history, and clinical signs. Tests should be selected and interpreted based on their relative sensitivity and specificity for specific conditions. It is essential to use effective testing methods for the etiologies of concern. This article reviews the plethora of diagnostic techniques available for fecal assessment. Indications, limitations, and issues of specimen handling for each technique are discussed. The optimal approach to the diagnosis of some common parasites, pathogens, abnormalities of flora, and metabolic conditions are covered. PMID- 14738203 TI - Diagnosis and management of reflux esophagitis. AB - Esophagitis has generally been considered rare in dogs and cats. However, it may not be as uncommon as previously thought due to the increased awareness of the disease and the increasing availability of endoscopy. Esophagitis can be caused by gastroesophageal reflux, trauma, foreign bodies, ingestion of caustic substances, structural abnormalities (ie, hiatal hernia, neoplasms), and chronic vomiting. This article will focus on the diagnosis and treatment of esophagitis secondary to gastroesophageal reflux. This is a diagnosis based on clinical signs, exclusion of other causes of esophagitis, and typical radiographic, endoscopic, and histopathologic findings. PMID- 14738204 TI - Current concepts in the treatment of canine chronic hepatitis. AB - Chronic hepatitis is a common disorder in dogs seen by general practitioners. Several new drugs have been marketed for treating this disease. Unfortunately, there are few controlled studies that examine the efficacy of these medications for the treatment of canine chronic hepatitis. A rational therapeutic approach can be implemented based on histopathologic findings of a liver biopsy. A liver biopsy is essential for establishing a definitive diagnosis and guiding the optimal therapy. The biopsy allows characterizing the inflammatory process, quantitating hepatic copper concentrations, and determining if fibrosis is present. Copper associated hepatopathy is treated with zinc and copper chelators. Idiopathic chronic hepatitis is thought to be immune mediated. The treatment of idiopathic chronic hepatitis consists of controlling inflammation (prednisone, azathioprine), reversing fibrosis (colchicine), and protecting against oxidant damage (vitamin E, ursodeoxycholic acid, S-adenosylmethionine). The prognosis for chronic hepatitis is quite variable. Dogs with end-stage disease have a poor prognosis, while dogs diagnosed earlier can have a mean survival of years. Early diagnosis and intervention are key to the successful treatment of dogs with chronic hepatitis. PMID- 14738205 TI - Diagnostic approaches to acute pancreatitis. AB - Acute pancreatitis is a challenging medical condition, with the potential for high mortality and comorbidity in companion animal patients. The anatomic position and relationships of the pancreas make direct observation of pancreatic pathology difficult, so the diagnosis of pancreatic disease is typically based on the measurement of biochemical markers in serum and imaging studies of the abdomen. The digestive enzymes typically used in the diagnosis of pancreatitis, amylase and lipase, have poor sensitivity and specificity. Recently, pancreas specific lipases have been purified from dogs and cats, leading to the availability of specific immunoassays for these proteins that show great promise in the diagnosis of acute pancreatitis. After the diagnosis of acute pancreatitis is established, there is a need to assess the severity of the disease to give an accurate prognosis for the patient. This article will review currently available methods used to diagnose acute pancreatitis and the ability of these methods to assess accurately the severity of the disease. PMID- 14738206 TI - The advantages and disadvantages of endoscopy. AB - Endoscopy is generally a very safe and effective tool in the diagnosis and therapy of various gastrointestinal (GI) disorders, and must be used in conjunction with other diagnostic modalities. Endoscopy should not be a substitute for a complete work-up. There are many advantages of endoscopy, including minimal morbidity and mortality, and the sensitivity of this modality in the diagnosis of mucosal disorders of the GI tract. However, complications may occur, and there are limitations to endoscopy. This article will provide an overview of when to choose GI endoscopy and when other procedures might provide more information. PMID- 14738207 TI - Maintenance of gastrointestinal endoscopes. AB - Endoscopes are used for visualization and biopsy of gastrointestinal lesions, as well as therapeutic procedures, such as foreign body retrieval. In the past, they were primarily used in large institutional settings where specialized personnel could focus on their maintenance. Today, they are becoming increasingly common in general practice. The maintenance of gastrointestinal endoscopes in the veterinary setting involves many challenges, including safe handling, reprocessing, and storage. Meeting these challenges requires well-trained personnel and strict protocols. Reprocessing, which includes cleaning and disinfection, offers the greatest challenge. The complex structure of flexible endoscopes, particularly the long, narrow channels, makes them difficult to clean. Gastrointestinal endoscopes operate in a contaminated environment, exposing them to high levels of organic matter and bacteria. High-level disinfection is necessary for infection control, but liquid germicides available for endoscope disinfection can be toxic to medical personnel and patients. PMID- 14738208 TI - Health effects of acute exposure to air pollution. Part I: Healthy and asthmatic subjects exposed to diesel exhaust. AB - The purpose of this study was to assess the impact of short-term exposure to diluted diesel exhaust on inflammatory parameters in human airways. We previously exposed control subjects for 1 hour to a high ambient concentration of diesel exhaust (particle concentration 300 pg/m3--a level comparable with that found in North Sea ferries, highway underpasses, etc). Although these exposures did not have any measurable effect on standard indices of lung function, there was a marked neutrophilic inflammatory response in the airways accompanied by increases in blood neutrophil and platelet counts. Endothelial adhesion molecules were upregulated, and the expression of interleukin 8 messenger RNA (IL-8 mRNA*) was increased in a pattern consistent with neutrophilia. Individuals with asthma have inflamed airways and are clinically more sensitive to air pollutants than are control subjects. The present study was designed to assess whether this clinical sensitivity can be explained by acute neutrophilic inflammation or an increase in allergic airway inflammation resulting from diesel exhaust exposure. For this study, we used a lower concentration of diesel exhaust (100 microg/m3 PM10) for a 2-hour exposure. At this concentration, both the control subjects and those with asthma demonstrated a modest but statistically significant increase in airway resistance following exposure to diesel exhaust. This increase in airway resistance was associated with an increased number of neutrophils in the bronchial wash (BW) fluid obtained from control subjects (median after diesel exhaust 22.0 vs median after air 17.2; P = 0.015), as well as an increase in lymphocytes obtained through bronchoalveolar lavage (BAL) (15.0% after diesel exhaust vs 12.3% after air; P = 0.017). Upregulation of the endothelial adhesion molecule P-selectin was noted in bronchial biopsy tissues from control subjects (65.4% of vessels after diesel exhaust vs 52.5% after air). There was also a significant increase in IL-8 protein concentrations in BAL fluid and IL-8 mRNA gene expression in the bronchial biopsy tissues obtained from control subjects after diesel exhaust exposure (median IL-8 expression 65.7% of adenine phosphoribosyl transferase [APRT] gene expression value after diesel exhaust vs 51.0% after air; P = 0.007). There were no significant changes in total protein, albumin, or other soluble inflammatory markers in the BW or BAL fluids. Red and white blood cell counts in peripheral blood were unaffected by diesel exhaust exposure. Airway mucosal biopsy tissues from subjects with mild asthma (defined as forced expiratory volume in 1 second [FEV1] greater than or equal to 70% of the predicted value) showed eosinophilic airway inflammation after air exposure compared with the airways of the corresponding control subjects. However, among the subjects with mild asthma, diesel exhaust did not induce any significant change in airway neutrophils, eosinophils, or other inflammatory cells; cytokines; or mediators of inflammation. The only clear effect of diesel exhaust on the airways of subjects with asthma was a significant increase in IL-10 staining in the biopsy tissues. This study demonstrated that modest concentrations of diesel exhaust have clear-cut inflammatory effects on the airways of nonasthmatic (or control) subjects. The data suggest a direct effect of diesel exhaust on IL-8 production leading to upregulation of endothelial adhesion molecules and neutrophil recruitment. Despite clinical reports of increased susceptibility of patients with asthma to diesel exhaust and other forms of air pollution, it does not appear that this susceptibility is caused either directly by induction of neutrophilic inflammation or indirectly by worsening of preexisting asthmatic airway inflammation. The increased level of IL 10 after diesel exhaust exposure in airways of subjects with asthma suggests that this pollutant may induce subtle changes in airway immunobiology. This is an important topic for further investigation. Other possible explanations for the apparent lack of response to diesel exhaust among subjects with asthma include (1) the time course of the response to diesel may differ from the response to allergens, which peaks 6 to 8 hours after exposure; (2) a different type of inflammation may occur that was not detectable by the standard methods used in this study; and (3) the increased sensitivity of patients with asthma to particulate air pollution may reflect the underlying bronchial hyperresponsiveness found in asthma rather than any specific increase in inflammatory responses. PMID- 14738209 TI - Health effects of acute exposure to air pollution. Part II: Healthy subjects exposed to concentrated ambient particles. AB - The purpose of this study was to assess the impact of short-term exposure to concentrated ambient particles (CAPs*) on lung function and on inflammatory parameters in blood and airways of healthy human subjects. Particles were concentrated from the ambient air in Chapel Hill, North Carolina, using a Harvard/EPA (US Environmental Protection Agency) ambient fine particle concentrator (HAPC). Each of 38 subjects was exposed either to filtered air (n = 8) or to CAPs (n = 30) for two hours, during which all subjects intermittently exercised. Blood was obtained immediately before and 18 hours after exposure. Also at 18 hours after exposure, viable bronchial biopsy tissues and lavage samples were obtained from 10 CAPs-exposed and 7 control subjects by fiberoptic bronchoscopy. To balance these two groups, additional biopsy tissues were obtained from 4 control subjects participating in an identical protocol for another study. For the CAPs-exposed group, the concentration of particulate matter measuring 2.5 microm or less in aerodynamic diameter (PM2.5) in the exposure aerosols varied from 23.1 to 311.1 microg/m3; for the filtered air group, mean particle concentration was 2.9 microg/m3. For comparative analyses, the CAPs-exposed subjects were separated into three tertiles on the basis of the final concentration of particles to which they were exposed. Lung function, assessed by spirometry and plethysmography before and immediately after exposure, was unaffected by CAPs. Of the inflammatory parameters studied in blood, subjects exposed to CAPs showed mean increases in fibrinogen of 40 to 48 mg/dL with no obvious differentiation by dose, whereas subjects exposed to filtered air showed no change; red and white blood cell counts were unaffected by CAPs exposure. In bronchoalveolar lavage fluid from CAPs-exposed subjects, neutrophils showed a dose-dependent increase both when analyzed as an absolute cell count and as a percentage of total lavaged cells. Bronchial biopsy tissues from 10 CAPs-exposed subjects and 11 control subjects did not show any consistent effect of CAPs exposure on cell counts or adhesion molecule expression. We conclude that CAPs induced a modest degree of airway inflammation as judged by lavage, but this effect was not reflected in biopsy tissues from proximal airways. This discrepant finding may mean that the inflammatory effect of CAPs occurs in more distal airways or that the health effects of PM are driven by processes other than those investigated in this study. PMID- 14738210 TI - Controlled exposures of healthy and asthmatic volunteers to concentrated ambient particles in metropolitan Los Angeles. AB - Epidemiologic studies report health effects associated with exposure to ambient particulate matter (PM), but underlying biologic mechanisms remain unclear. We evaluated pulmonary and systemic effects in twelve healthy human adult and twelve asthmatic volunteers exposed once for 2 hours in a whole-body chamber to approximately 200 microg/m3 concentrated ambient particles (CAPs) in the fine (< 2.5 microm) size range and once to filtered air. Neither healthy nor asthmatic subjects showed significant changes in symptoms, spirometry, or routine hematologic measurements attributable to CAPs exposure compared with filtered air. Both groups showed CAPs-related (1) decreases of columnar cells in induced sputum after exposure, (2) increases in certain blood mediators of inflammation (ie, soluble intercellular adhesion molecule 1 [ICAM-1] and interleukin [IL] 6 [marginally significant in asthmatic subjects only]), and (3) parasympathetic stimulation of heart rate variability (HRV). In the asthmatic group, systolic blood pressure modestly increased during filtered air exposure and decreased during CAPs exposure, whereas the pattern was reversed in the healthy group. In summary, this study measured a large number and wide range of biologic endpoints on a relatively small number of healthy and asthmatic volunteers and found few biologic endpoints that responded to CAPs and filtered air exposures with significantly different values. However, observed changes in some mediators of inflammation in blood and changes in HRV were consistent with PM-related effects reported from epidemiologic studies. They suggest that exposure to concentrated PM 2.5 pm or smaller in aerodynamic diameter (PM2.5) tends to elicit more systemic than pulmonary effects. This investigation is one of the first to apply concentrator-exposure technology in a high-risk group (subjects with asthma). Further studies of responses to CAPs that involve other biologic endpoints, other PM size modes, and subjects with other risk factors are needed. PMID- 14738211 TI - Let's ask them: a national survey of definitions of quality of life and its enhancement among people aged 65 and over. AB - This study aimed to explore older peoples' definitions of, and priorities for, a good quality of life for themselves and their peers. Nine hundred and ninety-nine people aged 65 and over, living at home in Britain, were interviewed for the study. Good social relationships were the most commonly mentioned constituent that gave respondents' lives quality (mentioned by 81 percent). Other important factors were social roles and activities, health, psychological outlook and well being, home and neighborhood, finances, and independence. Poor health was most often mentioned as taking quality away from life (by 50 percent). Social relationships and health were judged to be the most important areas. Having health and enough money were the two most frequently mentioned things that would improve the quality of their own lives and those of their peers (though in different order of magnitude). The need for dynamic, multidimensional, and integrated models of quality of life in older age is suggested by these results. PMID- 14738212 TI - The relationship between mastery and depression among Japanese family caregivers. AB - The present study examined the relationship between caregiver mastery and depressive symptoms among family stroke caregivers in western Japan (N = 100). Family caregivers were identified from a sample of rehabilitation hospitals; participation rate was 100 percent for all eligible caregivers. Caregivers with high mastery were found to have significantly fewer depressive symptoms than low mastery caregivers and were significantly more likely to use a respite caregiver. They also reported significantly less burden, yet paradoxically were significantly more likely to rate their care-recipients as more functionally dependent on them. Caregiver age, health status, and caregiving duration did not relate to mastery. However, men had a significantly higher sense of mastery. In general, findings parallel those for Western family caregivers, although mean mastery scores for Japanese caregivers were lower than those reported for American family caregivers. PMID- 14738213 TI - Looking for signs of Alzheimer's disease. AB - This study examined the correlates of symptom-seeking behavior for Alzheimer's disease (AD) among middle-aged persons. Symptom seeking, the tendency to search for signs of disease, is one manifestation of an individual's concern about developing AD. The data were obtained from a survey of two subsamples of 40-60 year old adults: 1) 108 adult children with a living parent with a diagnosis of probable AD; and 2) 150 adults in a matched group with no parental history of AD. Bivariate and multivariate analyses were used to identify significant predictors of symptom seeking, which was measured by a composite index comprised of responses from three questions about checking for signs of AD, interpreting signs as symptoms of AD, and soliciting external validation for concerns. Four clusters of predictors were examined: memory assessment, AD experience, sociodemographics, and well-being. Within these clusters, the constellations of significant predictors varied by subsample, but the most robust predictors were aspects of subjective assessments of memory functioning and AD experience. An understanding of the correlates of symptom seeking for AD has implications for early detection of the disease as well as identifying populations under stress from excessive worry about their own future health. PMID- 14738214 TI - The meaning of the future for the oldest old. AB - The aim of the study was to highlight the oldest old people's view of their future from a perspective of philosophy of life. Data was collected by means of life story interviews with 15 persons. The analysis was performed by utilizing a phenomenological hermeneutic method and the interpretation was guided by the conceptual framework of philosophy of life as designated by Jeffner (1988). The following themes emerged: future seen as everyday life; future-oriented values; and thoughts about life and death. The oldest old were found to view their future in ways that ranged from a tangible positive approach via a wait-and-see policy to a negative approach. Their perception of their future implied two different time perspectives, their immediate future and a more long-term perspective of the future. Furthermore, the future was experienced on three different levels, the individual level, the intergenerational level, and the metaphysical level. PMID- 14738215 TI - Epitope identification and discovery using phage display libraries: applications in vaccine development and diagnostics. AB - Antigenic epitopes are the part (contact points) of an antigen involved in specific interaction with the antigen-binding site (the paratope) of an antibody or a T-cell receptor. Detailed analysis of epitopes is important both for the understanding of immunological events and for the development of more effective vaccine and diagnostic tools for various diseases. Identification and characterization of epitopes is a complex process. Although various methods have been developed in this area, there still lacks a simple common approach which can be applied to all epitopes. Since its first introduction more than a decade ago, phage display technology has made a major impact in this area of research. With the exponential growth in this area, it is impractical to review the entire literature detailing all possible applications. Instead, this review aims to focus on specific applications related to the discovery and identification of epitopes which have potential as vaccine candidates or can be used in disease diagnosis. PMID- 14738216 TI - Dendritic cells in tumor immunology and immunotherapy. AB - Despite rapid advances in cancer therapeutics, relapsed disease due to failed immunosurveillance remains a major problem in many cancers. Dendritic cells are recognized as key to the induction of immune responses to cancer and intensive study is underway to facilitate their use in cancer immunotherapy. In initial clinical trials, dendritic cell preparations were, with the benefit of hindsight, largely sub-optimal, yet encouraging results have been seen. The challenge now is to expand our knowledge of the interactions between tumors and the immune system, through basic scientific research and coordinated large-scale clinical studies. This review focuses on the anti-tumor immune response, human dendritic cell biology and the results of recent clinical studies of dendritic cell immunotherapy for cancer. PMID- 14738217 TI - Neutralising antibody, CTL and dendritic cell responses to hepatitis C virus: a preventative vaccine strategy. AB - Hepatitis C virus (HCV) accounts for the majority of cases of transfusion acquired hepatitis and hepatitis transmitted by injecting drug use. The patients who do not clear the infection become chronic carriers of HCV and form a reservoir of infection within human populations. Furthermore, these carriers are at serious risk of developing cirrhosis of the liver and hepatocellular carcinoma. The disease is of major concern in developing as well as in developed countries and yet there are no vaccines against HCV, treatment is confined to the use of chemotherapy which is expensive and not always effective. The major obstacle in vaccine development is a limited understanding of the type of immune response that is necessary for viral clearance and the occurrence of various genotypes and quasispecies of HCV. The problems are further compounded by difficulties in growing the virus in vitro and the lack of a suitable and economical animal model. PMID- 14738218 TI - Vaccine development for group A streptococcus infections and associated disease. AB - Group A streptococcus (GAS) is responsible for a number of diseases ranging from uncomplicated pharyngitis through to life-treating invasive and post-infectious diseases such as necrotizing fasciitis and rheumatic heart disease. GAS associated diseases occur globally and are serious problems in many developing nations and indigenous populations of many developed nations. This, and the resurgence in industrialized countries, and increased virulence of GAS in the 1980s highlight the need of cost-effective control strategies. Here we highlight the GAS diseases that are still a problem in many populations and discuss potentially useful strategies to combat GAS infections and disease. PMID- 14738219 TI - Vaccines and vaccine strategies against HIV. AB - The HIV/AIDS pandemic is a global emergency and a preventive HIV vaccine is urgently needed. HIV has, however, proved a difficult pathogen to vaccinate against. This is largely because HIV has a very high mutation rate and can escape immune responses, it has a latent stage where it can rest silently integrated into host DNA, and neutralising antibodies that can neutralise diverse field strains have so far proved difficult to induce. There is however, considerable evidence now that HIV-specific CD4 and CD8 T cells can provide partial control of HIV replication and delay or prevent disease. Technologies to quantify and analyse HIV-specific T cells have advanced recently, and in particular ELISpot, intracellular cytokine staining and tetramer studies have provided clear analyses of the ability of HIV vaccines to induce T cell responses. The use of pools of overlapping HIV peptides as in vitro antigens has also provided a standardised reagent for accurate measurement of T cell responses. HIV protein vaccines have not induced broad neutralising antibodies or T cell responses and failed to protect humans in the only phase III efficacy trial yet completed. Viral vectors, such as canarypox, engineered to express HIV genes, have induced HIV-specific CD8 T cell responses in a minority of subjects in phase II trials and are proceeding to human efficacy trials. Currently, the most effective method of inducing CD8+ CTL immunity in non-human primates utilises priming with naked plasmid DNA and then boosting with recombinant viral vectors both encoding various parts of the HIV genome. Such vaccines have induced non-sterilising immunity to virulent Simian/Human immunodeficiency virus exposure in macaques and have entered phase I trials. Multiple other approaches are also being evaluated in what has become a global effort for a vaccine to prevent AIDS. Although an HIV vaccine is still a long way off, there is reason to be optimistic that a vaccine to prevent AIDS will eventually be developed. PMID- 14738220 TI - Alternatives to conventional vaccines--mediators of innate immunity. AB - Vaccines have been described as "weapons of mass protection". The eradication of many diseases is testament to their utility and effectiveness. Nevertheless, many vaccine preventable diseases remain prevalent because of political and economic barriers. Additionally, the effects of immaturity and old age, therapies that incapacitate the adaptive immune system and the multitude of strategies evolved by pathogens to evade immediate or sustained recognition by the mammalian immune system are barriers to the effectiveness of existing vaccines or development of new vaccines. In the front line of defence against the pervasiness of infection are the elements of the innate immune system. Innate immunity is under studied and poorly appreciated. However, in the first days after entry of a pathogen into the body, our entire protective response is dependant upon the various elements of our innate immune repertoire. In spite of its place as our initial defence against infection, attention is only now turning to strategies which enhance or supplement innate immunity. This review examines the need for and potential of innate immune therapies. PMID- 14738221 TI - Does "A" stand for alveolization? PMID- 14738222 TI - Improvement in volitional tests of muscle function alone may not be adequate evidence that inspiratory muscle training is effective. PMID- 14738223 TI - The interface: crucial for successful noninvasive ventilation. PMID- 14738224 TI - Chronic eosinophilic pneumonia after radiation therapy for breast cancer. AB - The priming of bronchiolitis obliterans organising pneumonia by radiation therapy (RT) to the breast is now a well recognised syndrome. This study describes the occurrence of chronic eosinophilic pneumonia following RT after surgery for breast cancer in five female patients, with a mean age of 68 yrs (range 49-77). All patients had a history of asthma and/or allergy. At the onset of eosinophilic pneumonia, all patients were symptomatic. Chest radiograph showed pulmonary infiltrates, unilateral and limited to the irradiated lung in three patients, and bilateral in two. Pulmonary opacities were migratory in one patient. All patients had blood eosinophilia >1.0 10(9) x L(-1) and/or eosinophilia >40% at bronchoalveolar lavage differential cell count. The median time interval between the end of radiation therapy and the onset of eosinophilic pneumonia was 3.5 months (range 1-10). All patients rapidly improved with oral corticosteroids without sequelae. Relapse occurred in two patients after treatment withdrawal. Priming of alveolitis by radiation therapy to the breast might promote either bronchiolitis obliterans organising pneumonia or chronic eosinophilic pneumonia, with the latter depending on genetic or acquired characteristics of patients and/or further stimulation that may trigger a T-helper cell type 2 form of lymphocyte response, especially in patients with asthma or other atopic manifestations. PMID- 14738225 TI - Nuclear YB-1 expression as a negative prognostic marker in nonsmall cell lung cancer. AB - The human Y-box binding protein, YB-1, is a multifunctional protein that regulates gene expression. Nuclear expression of YB-1 has been associated with chemoresistance and poor prognosis of tumour patients. Representative samples from autopsied material of primary tumours from 77 patients with NSCLC were investigated by immunohistochemistry for subcellular distribution of YB-1 and p53, in order to evaluate the prognostic role of nuclear expression of YB-1. Cytoplasmic YB-1 expression was found in all tumour samples, whereas nuclear expression was only observed in 48%. There was no correlation with histological classification, clinical parameters or tumour size, stage and metastasis status. However, patients with positive nuclear YB-1 expression in tumours showed reduced survival times when compared with patients without nuclear expression. Including information about the histology and mutational status for p53 increased the prognostic value of nuclear YB-1. Patients with nuclear YB-1 expression and p53 mutations had the worst prognosis (median survival 3 months), while best outcome was found in patients with no nuclear YB-1 and wildtype p53 (median survival 15 months). This suggests that the combined analysis of both markers allows a better identification of subgroups with varying prognosis. Nuclear expression of Y-box binding protien seems to be an independent prognostic marker. PMID- 14738226 TI - Retinoic acid induces alveolar regeneration in the adult mouse lung. AB - Recent data suggests that exogenous retinoic acid (RA) can induce alveolar regeneration in a mouse and a rat model of experimental emphysema and disrupted alveolar development. This may be because RA is required during normal alveolar development and the subsequent provision of RA reawakens the gene cascades used during development. Here, additional evidence that RA is required during alveologenesis in the mouse is provided by showing that disulphiram disrupts this process. A further model of disrupted alveolar development using dexamethasone administered postnatally is then described, and it is further shown that RA administered to these adult mice restores the lung architecture to normal. Alveolar regeneration with retinoic acid may therefore be an important novel therapeutic approach to the treatment of respiratory diseases characterised by a reduced gas-exchanging surface area, such as bronchopulmonary dysplasia and emphysema. PMID- 14738227 TI - The 6-min walk distance: change over time and value as a predictor of survival in severe COPD. AB - The 6-min walk distance (6MWD) is used to evaluate the functional capacity of patients with chronic obstructive pulmonary disease (COPD). The change in 6MWD over time and its correlation with changes in spirometry and survival are unclear. Patients (n=198) with severe COPD and 41 age-matched controls were followed for 2 yrs, and anthropometrics, spirometry, 6MWD and comorbidities were measured. The 6MWD decreased in the COPD group from 238 +/- 107 m to 218 +/- 112 m (-26 +/- 37 m x yr(-1)), and increased in the control group from 532 +/- 82 m to 549 +/- 86 m (12 +/- 25 m x yr(-1)). In both groups, there was a poor correlation with changes in forced expiratory volume in one second (FEV1). Nonsurvivors in the COPD group (42%) had a more pronounced change in the 6MWD ( 40 versus -22 m x yr(-1)) but a similar change in FEV1 (118 versus 102 mL x yr( 1)). The 6MWD independently predicted survival, after accounting for age, body mass index, FEV1 and comorbidities. In severe chronic obstructive pulmonary disease, the 6-min walk distance predicts mortality better than other traditional markers of disease severity. Its measurement is useful in the comprehensive evaluation of patients with severe disease. PMID- 14738228 TI - Acute effects of inspiratory pressure support during exercise in patients with COPD. AB - The aim of the present study was to evaluate the acute effects of inspiratory pressure support (IPS) of 5 cmH2O (IPS5) and 10 cmH2O (IPS10) on exercise endurance. Forty-five patients with COPD (mean forced expiratory volume in one second (FEV1) = 39 +/- 14% pred) performed three constant-load endurance tests on a cycle ergometer at 75% of maximal workload. One test was without IPS, one test with IPSs, and one with IPS10. No statistically significant difference was found in exercise endurance between tests without IPS and IPS5 (4.2 +/- 2.6 versus 4.4 +/- 2.9 min). In contrast, IPS10 resulted in a statistically significant increase in endurance compared with exercise without IPS (6.3 +/- 6.7 versus 4.2 +/- 2.6 min), as well as compared with exercise with IPS5 (6.3 +/- 6.7 versus 4.4 +/- 2.9 min). A wide scatter in individual responses to IPS was found, ranging from a deterioration of 1.6 min (-36%) to an improvement of 16.3 min (+445%). In only 15 patients, the increase in endurance exceeded the upper limit of the 95%, confidence interval. Stepwise multiple regression analysis showed that maximal inspiratory pressure was the most important determinant of the increase in exercise endurance due to the application of IPS10. It was concluded that in contrast to inspiratory pressure support of 5 cmH2O, the application of inspiratory pressure support of 10 cmH2O during exercise resulted in a statistically significant improvement in exercise endurance in patients with COPD compared with exercise without inspiratory pressure support. However, on an individual basis, large differences in responses were found. Inspiratory muscle weakness was revealed as a determinant of improvement in exercise endurance due to the application of inspiratory pressure support of 10 cmH2O, explaining only 24% of the variance in outcome. PMID- 14738229 TI - Sleep-related hypoxaemia and excessive erythrocytosis in Andean high-altitude natives. AB - To determine whether nocturnal hypoxaemia contributes to the excessive erythrocytosis (EE) in Andean natives, standard polysomnographies were performed in 10 patients with EE and in 10 controls (mean haematocrit 76.6 +/- 1.3% and 5.4 +/- 0.8%, respectively) living at an altitude of 4,380 m. In addition, the effect of O2 administration for 1 h prior to sleep, and the relationship between the hypoxic/hypercapnic ventilatory response and the apnoea/hypopnoea index (AHI) during sleep were studied. Awake arterial oxygen saturation (Sa,O2) was significantly lower in patients with EE than in controls (83.7 +/- 0.3% versus 85.6 +/- 0.4%). In both groups, the mean Sa,O2 significantly decreased during sleep (to 80.0 +/- 0.8% in EE and to 82.8 +/- 0.5% in controls). The mean Sa,O2 values remained significantly lower in patients with EE than in controls at all times of the night, and patients with EE spent significantly more time than the controls with an Sa,O2 of <80%. There were no differences between the two groups in the number and duration of the apnoeas/hypopnoeas. None of these variables were affected by O2 administration. In both groups the AHI positively correlated with the hypercapnic ventilatory response. Andean natives undergo minor respiratory disorders during sleep. The reduction in oxygen saturation found in subjects with excessive erythrocytosis was small, yet consistent and potentially important, as it remained below the threshold known for the increase in erythropoietin stimulation. This may be an important factor promoting erythropoiesis, but its relevance needs to be further explored. PMID- 14738230 TI - Theophylline and acetazolamide reduce sleep-disordered breathing at high altitude. AB - A randomised, double-blind, placebo-controlled study was conducted to evaluate the effects of theophylline and acetazolamide in the treatment of sleep disordered breathing (SDB) after fast ascent to high altitude (3,454 m). The study was conducted at a high-altitude research laboratory and included 30 healthy male volunteers. Study medication was either oral slow release theophylline (2x250 mg x day(-1)), oral acetazolamide (2x250 mg x day(-1)) or a matched placebo tablet. Polysomnographic measurements were performed during two consecutive nights, and acute mountain sickness, pulse rate, oxyhaemoglobin saturation and arterial blood gases were assessed three times a day. Without active medication, the apnoea/hypopnoea index (AHI) was highly pathological (median 16.2 x h(-1), range 2-92). Both theophylline and acetazolamide normalised SDB (median AHI 2.5 x h(-1), range 0-11; 4.2 x h(-1), range 0-19, respectively) and reduced oxyhaemoglobin desaturations during sleep (median desaturation index placebo 41.5 x h(-1), range 6-114; acetazolamide 6.5 x h(-1), range 3-28; theophylline 8.5 x h(-1), range 3-32). Compared with the high amount of central apnoeas or hypopnoeas, the number of obstructive events during sleep was very low in all groups (<4 x h(-1)). In contrast to theophylline, acetazolamide significantly improved basal oxyhaemoglobin saturation during sleep (86.2 +/- 1.7% versus 81.0 +/- 3.0%). The authors conclude that both oral slow release theophylline and acetazolamide are effective to normalise high-altitude sleep disordered breathing. PMID- 14738231 TI - Economic arguments for the immediate management of moderate-to-severe obstructive sleep apnoea syndrome. AB - The objective of this study was to measure the impact of a 6-month delay in the diagnosis and treatment of patients with moderate obstructive sleep apnoea syndrome (OSAS) (apnoea/hypopnoea index (AHI) < 30) or severe OSAS (AHI > or = 30) on daytime sleepiness, cognitive functions, quality of life and healthcare expenditure (hospitalisations, medical visits, complementary tests, biological tests and drug prescriptions). In addition, this study aimed to analyse the incremental cost effectiveness ratios related to daytime sleepiness or quality of life following immediate introduction of treatment in these two populations. This study was conducted as a multicentre randomised controlled trial and carried out at two teaching hospitals in France. A total of 171 patients were followed for 6 months, with 82 patients randomised to group 1 "immediate polysomnography" and 89 in group 2 "polysomnography within 6 months". Patients with severe OSAS were deprived of a significant improvement of their daytime sleepiness (5.1 +/- 5.0 at the Epworth Sleepiness Scale score in group 1 versus 0.2 +/- 3.4 in group 2) and quality of life (12.4 +/- 13.3 at the Nottingham Health Profile score in group 1 versus 0.7 +/- 10.1 in group 2) during the waiting time. The impact of delayed management in subjects with less severe OSAS only concerned daytime sleepiness (1.1 +/- 3.3 in group 1 versus 0.3 +/- 4.3 in group 2). Delayed treatment did not affect cognitive functions or healthcare expenditure regardless of the severity of the disease. Incremental cost effectiveness ratios related to rapid introduction of treatment were significantly lower in the patients with more severe OSAS. These results provide fairly clear medical and economic arguments in favour of early management of patients with more severe forms of obstructive sleep apnoea syndrome. PMID- 14738232 TI - Maintenance of inspiratory muscle training in COPD patients: one year follow-up. AB - In most chronic obstructive pulmonary disease (COPD) patients, dyspnoea and functional exercise capacity may improve as a result of inspiratory muscle training (IMT). However, the long-term benefits of IMT have been investigated to a much lesser extent. The present study investigated the short-term and long-term benefits of IMT on inspiratory muscle performance (strength and endurance), exercise capacity and the perception of dyspnoea. Thirty-eight patients with significant COPD had 3 months of basic IMT and were then randomised into a group that received maintenance IMT for the next year, and a group that got training with very low load. Following the basic training there was a statistically significant increase in inspiratory muscle performance, 6-min walk test (6MWT), and a decrease in the dyspnoea. During the second stage of the study, the training group continued to maintain the improvement in all parameters, while there was already deterioration in the inspiratory muscle performance, exercise capacity and dyspnoea in the low intensity group during the 6-12 month period. The present study concludes that, in patients with significant chronic obstructive pulmonary disease, inspiratory muscle training results in improvement in performance, exercise capacity and in the sensation of dyspnoea. The benefits of 12-weeks of inspiratory muscle training decline gradually over 1 yr of follow up if maintenance training is not performed. PMID- 14738233 TI - Childhood factors that predict asthma in young adulthood. AB - Predicting adult asthma, using childhood characteristics, is important for advising on prognosis and, potentially, for secondary prevention. A novel use of multivariate likelihood ratios (LRs) to quantify prognosis is described here. Of 718 subjects of a community-based cohort, 575 (80%) members were recruited at age 8-10 yrs and were re-assessed 15-17 yrs later. At baseline, information about symptoms, spirometry, histamine challenge and skin-prick tests were collected. At follow-up "asthma symptoms" were defined as wheeze, sleep disturbance from asthma or inhaled steroid use within the previous year. LRs were calculated for significant predictors of this outcome. Shinkage factors were applied to yield multivariate LRs. Childhood characteristics that independently predicted asthma symptoms in adulthood were obstructive spirometry (adjusted (adj)LR 2.9, 95% confidence interval (CI) 1.3-6.5), airway hyperresponsiveness (adjLR 2.6, 95% CI 1.8-3.7), atopy (adjLR 2.0, 95% CI 1.5-2.7), recent wheeze (adjLR 1.9, 95% CI 1.5 2.5) and being female (adjLR 1.29, 95% CI 0.8-2.1). Children with all five characteristics had a cumulative LR of 36.9 for asthma symptoms in adulthood. Most adults who had asthma symptoms did not have manifestations of asthma as children. However, the presence of obstructive spirometry, airway hyperresponsiveness and atopy in childhood identifies individuals with increased likelihood of having asthma in adulthood. Cumulative likelihood ratios are more valuable than odds ratios for quantifying risk in individuals and for identifying people with most to gain from preventive interventions. PMID- 14738234 TI - Formoterol protects against platelet-activating factor-induced effects in asthma. AB - Platelet-activating factor (PAF) is an inflammatory mediator that provokes neutropaenia, bronchoconstriction and gas exchange defects due to exudation of bulk plasma within the airways. While the inhibitory effects of short-acting beta2-agonists on PAF-induced disturbances have been consistently shown, those of long-acting beta2-agonists are less convincing. To further explore the mechanisms involved in PAF challenge in asthma, 12 patients (forced expiratory volume in one second, 90 +/- 4% predicted) were investigated 2 h after inhaled formoterol (18 microg), in a double-blind, placebo-controlled, crossover design following PAF (18 microg) inhalation. Compared with the placebo, at 5 min, premedication with formoterol reduced PAF-induced cough and dyspnoea, and attenuated increased respiratory system resistance (by 67%) and arterial deoxygenation (by 50%). Likewise, ventilation-perfusion (V'A/Q') inequality improved, as reflected by the dispersion of pulmonary blood flow (by 63%) and an overall index of V'A/Q' heterogeneity (by 71%). In contrast, PAF-induced facial flushing, neutropaenia and subsequent rebound neutrophilia remained unchanged. The improvement in gas exchange abnormalities shown after platelet-activating factor in patients with asthma pretreated with formoterol at the recommended clinical dose may reflect, in addition to its class effects, an anti-exudative effect of formoterol in the airways. PMID- 14738235 TI - Dual tachykinin NK1/NK2 antagonist DNK333 inhibits neurokinin A-induced bronchoconstriction in asthma patients. AB - Inhalation of neurokinin A (NKA) causes bronchoconstriction in patients with asthma. In vitro both tachykinin NK1 and NK2 receptors can mediate airway contraction. In this study the authors examined the effects of a single dose of the dual tachykinin NK1/NK2 receptor antagonist, DNK333, on NKA-induced bronchoconstriction in asthma. A total of 19 male adults with mild asthma completed a randomised, double-blind, placebo-controlled crossover trial. Increasing concentrations of NKA (3.3x10(-9) to 1.0x10(-6) mol x mLP(-1)) were inhaled at 1 and 10 h intervals after a single oral dosing with either DNK333 (100 mg) or a placebo. It was observed that DNK333 did not affect baseline lung function but did protect against NKA-induced bronchoconstriction in those patients. The mean log10 provocative concentration causing a 20% fall in forced expiratory volume in one second for NKA was -5.6 log10 mol x mL(-1) at 1 h after DNK333 treatment and -6.8 log10 mol x mL(-1) after placebo. This was equivalent to a difference of 4.08 doubling doses, which decreased to a difference of 0.90 doubling doses 10 h after treatment. The results shown in this report indicate that DNK333 blocks neurokinin A-induced bronchoconstriction in patients with asthma. PMID- 14738236 TI - Different airway inflammatory responses in asthmatic and healthy humans exposed to diesel. AB - Particulate matter (PM) pollution adversely affects the airways, with asthmatic subjects thought to be especially sensitive. The authors hypothesised that exposure to diesel exhaust (DE), a major source of PM, would induce airway neutrophilia in healthy subjects, and that either these responses would be exaggerated in subjects with mild allergic asthma, or DE would exacerbate pre existent allergic airways. Healthy and mild asthmatic subjects were exposed for 2 h to ambient levels of DE (particles with a 50% cut-off aerodynamic diameter of 10 microm (PM10) 108 microg x m(-3)) and lung function and airway inflammation were assessed. Both groups showed an increase in airway resistance of similar magnitude after DE exposure. Healthy subjects developed airway inflammation 6 h after DE exposure, with airways neutrophilia and lymphocytosis together with an increase in interleukin-8 (IL-8) protein in lavage fluid, increased IL-8 messenger ribonucleic acid expression in the bronchial mucosa and upregulation of the endothelial adhesion molecules. In asthmatic subjects, DE exposure did not induce a neutrophilic response or exacerbate their pre-existing eosinophilic airway inflammation. Epithelial staining for the cytokine IL-10 was increased after DE in the asthmatic group. Differential effects on the airways of healthy subjects and asthmatics of particles with a 50% cut-off aerodynamic diameter of 10 microm at concentrations below current World Health Organisation air quality standards have been observed in this study. Further work is required to elucidate the significance of these differential responses. PMID- 14738237 TI - Prevalence of asthma and exhaled nitric oxide are increased in bleachery workers exposed to ozone. AB - The aims of the present study were to determine whether exposure to high peaks of ozone resulted in an increased prevalence of asthma or respiratory symptoms among bleachery workers and whether nitric oxide (NO) was elevated in the exhaled air of these workers. Bleachery workers (n=228) from three Swedish pulp mills who had been exposed to ozone, together with 63 unexposed control subjects, were investigated by means of spirometry, Phadiatop, exhaled and nasal NO and answers to a questionnaire concerning respiratory symptoms and exposure. Exposure to an ozone peak that gave rise to respiratory symptoms was defined as a "gassing". Bleachery workers reporting four or more gassings involving ozone had an increased prevalence of adult-onset asthma, wheeze, and current asthma symptoms. They also had a higher median concentration of exhaled NO in comparison with those who reported no such gassings (19.2 versus 15.7 parts per billion). No such associations were found in respect of nasal NO. The results from this study show that bleachery workers who have been repeatedly exposed to ozone gassings have an increased prevalence of adult-onset asthma. The results also indicate exhaled nitric oxide may be a marker of airway inflammation in bleachery workers who have been exposed to high peaks of ozone. PMID- 14738238 TI - Progressive damage on high resolution computed tomography despite stable lung function in cystic fibrosis. AB - For effective clinical management of cystic fibrosis (CF) lung disease it is important to closely monitor the start and progression of lung damage. The aim of this study was to investigate the ability of high-resolution computed tomography (HRCT) scoring systems and pulmonary function tests (PFT) to detect changes in lung disease. CF children (n=48) had two HRCT scans in combination with two PFT 2 yrs apart. Their scans were scored using five scoring systems (Castile, Brody, Helbich, Santamaria and Bhalla). "Sensitivity" was defined as the ability to detect disease progression. In this group of children, HRCT scores worsened. PFT remained unchanged or improved. Of the HRCT parameters, mucous plugging and the severity, extent and peripheral extension of bronchiectasis worsened significantly. Relationships between changes in HRCT scores and PFT were weak. Substantial structural lung damage was evident in some children who had normal lung function. These data show that high-resolution computed tomography is more sensitive than pulmonary function tests in the detection of early and progressive lung disease, and suggest that high-resolution computed tomography may be useful in the follow up of cystic fibrosis children and as an outcome measure in studies that aim to reduce lung damage. PMID- 14738239 TI - Case-control study of Stenotrophomonas maltophilia acquisition in cystic fibrosis patients. AB - The aims of this case-control study were to describe the characteristics of cystic fibrosis (CF) patients who isolated Stenotrophomonas maltophilia in sputum, to determine risk factors for acquisition, to assess persistence of the organism and clinical outcomes postacquisition. Data were collected from 1991 1999. CF patients and controls (who had never isolated S. maltophilia) were matched for age (+/- 1 yr), sex and forced expiratory volume in one second (+/- 10%). Data were collected from the year prior and for 2 yrs postacquisition of S. maltophilia. The prevalence of S. maltophilia increased from 3.3% to 15%. Factors associated with S. maltophilia acquisition were more than two courses of intravenous antibiotics, isolation of Aspergillus fumigatus in sputum and oral steroid use. The effect of A. fumigatus was independent of steroid use. Clinical status did not change postacquisition. The majority of patients cleared the organism from the sputum. Long-term infection or an accelerated deterioration in lung function or nutrition is not likely post-Stenotrophomonas maltophilia acquisition in cystic fibrosis. This is the first documentation of an association between Aspergillus fumigatus isolation and Stenotrophomonas maltophilia acquisition in cystic fibrosis, independently of steroid therapy. PMID- 14738240 TI - Deaths in risk classes I-III: a measure of quality of care in patients hospitalised with CAP? AB - All deaths occurring in patients with community-acquired pneumonia in risk classes I-III were reviewed as a quality-of-care measure. The immediate and underlying causes of death were classified according to the World Health Organization protocol. Eleven (1.8%) of the 608 low-risk patients died. Three of the patients did not have pneumonia, one of whom was admitted with atypical pulmonary oedema due to stenosis of a prosthetic heart valve. Failure to include chronic lung disease in the severity-of-illness scoring system resulted in misclassification of seven patients. The most common underlying causes of death were pulmonary fibrosis at 27%, chronic obstructive lung disease at 18% and cancer at 27%. Respiratory failure was the immediate cause of death in 64% of patients, cardiac causes in 27%, and pneumonia in 9%. To conclude, the review of deaths in patients at low risk for mortality is useful for monitoring the quality of care received by patients who require admission for the treatment of community acquired pneumonia, and that the pneumonia-specific severity-of-illness scoring system results in misclassification of patients with chronic obstructive lung disease and pulmonary fibrosis. PMID- 14738241 TI - Pro- and anti-inflammatory responses in respiratory syncytial virus bronchiolitis. AB - Respiratory syncytial virus (RSV) bronchiolitis is an important cause of severe respiratory disease in infants. This study aimed to characterise changes in pulmonary pro- and anti-inflammatory responses in infants with RSV bronchiolitis over the course of the illness. On the day of intubation (Day 1) and the day of extubation (Day X), nonbronchoscopic bronchoalveolar lavage was performed on term and preterm infants ventilated for RSV bronchiolitis and on control infants on Day 1. Tumour necrosis factor (TNF)-alpha, soluble TNF receptor (sTNFR) and interleukin (IL)-6 messenger ribonucleic acid (mRNA) and protein were measured. Twenty-four infants, born at term and 23 infants born preterm with RSV bronchiolitis and 10 controls were recruited. TNF-alpha and IL-6 mRNA and protein in infants with bronchiolitis were greater than the control group on Day 1. In preterm infants, who were ventilated for longer than term infants, TNF-alpha and IL-6 proteins decreased between Day 1 and Day X. Concentrations of sTNFRs differed between groups on Day 1, but levels did not change between Day 1 and Day X. Large amounts of tumour necrosis factor-alpha and interleukin-6 in the respiratory syncytial virus-infected lung suggest important roles for these cytokines in the pathogenesis of respiratory syncytial virus bronchiolitis. The decrease in tumour necrosis factor-alpha and interleukin-6 protein in preterm infants may reflect the prolonged clinical course seen in these infants. PMID- 14738242 TI - Apoptosis and proliferation in lungs of ventilated and oxygen-treated preterm infants. AB - Apoptosis and proliferation and the effect of exogenous surfactant on these processes were investigated in the lungs of mechanically ventilated/oxygen treated preterm infants with respiratory distress syndrome and stillborn foetuses. Apoptotic and proliferation indices were determined in lung tissue sections from 27 ventilated/oxygen-treated preterm infants and 29 stillborn foetuses. The effect of exogenous surfactant on apoptosis and proliferation was studied in 16 ventilated preterm infants; 11 untreated infants served as control. Apoptotic and proliferating cells were identified by double labelling combining terminal deoxynucleotidyltransferase-mediated deoxyuridine triphosphate nick end labelling or Ki-67 with cell marker proteins. Pathways to cell death were explored by immunolabelling of cleaved caspases-3, -8 and -9. In the lungs of ventilated/oxygen-treated preterm infants, the numbers of apoptotic and proliferating cells increased significantly compared to the respective numbers in the lungs of stillborn foetuses. Apoptosis was detected in alveolar epithelial cells, whereas epithelial, endothelial and smooth muscle cells proliferated. Surfactant treatment reduced apoptosis induced by ventilation/oxygen-treatment; however, the decrease was not significant. Caspases-8 and -9 do not contribute to ventilation-induced apoptosis, whereas caspase-3 is involved. In conclusion, ventilation/oxygen-treatment induces epithelial cell apoptosis and proliferation of epithelial, endothelial and smooth muscle cells in the lungs of preterm infants. PMID- 14738243 TI - Mechanical ventilation of healthy rats suppresses peripheral immune function. AB - This study was designed to investigate the possible effect of injurious mechanical ventilation on peripheral immune function of healthy rats. Three ventilation strategies were compared: 1) low peak inspiratory pressure (PIP)/positive end-expiratory pressure (PEEP); 2) high PIP/PEEP; and 3) high PIP/zero PEEP (ZEEP). As a reference group, healthy, nonventilated, sham operated, anaesthetised rats were used. After 4 h, rats were sacrificed and macrophage inflammatory protein (MIP)-2 levels in lung and plasma were determined. Peripheral immune function was determined by measurement of splenic natural killer (NK) activity, mitogen-induced splenocyte proliferation and in vitro cytokine production. All immune measurements in the low PIP/PEEP group did not differ from the immune measurements in the reference group. High PIP strategies, irrespective of applied PEEP, enhanced MIP-2 levels in lung and plasma. NK cell activity, mitogen-induced splenocyte proliferation and MIP-2 and interleukin (IL)-10 production significantly decreased after high PIP/PEEP ventilation. In the high PIP/ZEEP-ventilated group, the decrease in splenocyte proliferation, MIP-2 and IL-10 production and NK cell activity was more pronounced and interferon-gamma production was also significantly lower than in the low PIP/PEEP group. These data show that high positive inspiratory pressure ventilation induces an inflammatory response in the lung, whereas at the same time the peripheral immune response is downregulated. Ventilator-induced peripheral immune suppression may contribute to poor outcome in acute respiratory distress syndrome patients. PMID- 14738244 TI - Dynamic dead space in face masks used with noninvasive ventilators: a lung model study. AB - The aim of this study was to determine what the influence of different designs of face masks and different noninvasive ventilator modes would be upon total dynamic dead space. Using a spontaneous breathing model, total dynamic dead space was measured when using 19 commercially available face masks and a range of ventilators in various ventilation modes. Total dynamic dead space during spontaneous ventilation was increased above physiological dead space from 32% to 42% of tidal volume by using face masks. The use of noninvasive ventilation modes such as bilevel and continuous positive airway pressure, with continuous pressure throughout the expiratory phase, reduced total dynamic dead space to approach physiological dead space with most face masks. Pressure assist and pressure support ventilation decreased total dynamic dead space to a lesser degree, from 42% to 39% of tidal volume. Face masks with expiratory ports over the nasal bridge resulted in beneficial flow characteristics within the face mask and nasal cavity, so as to decrease total dynamic dead space to less than physiological dead space from 42% to 28.5% of tidal volume. Exhaust ports over the nasal bridge in face masks effect important decreases in dynamic dead space provided positive pressure throughout the expiratory phase is used. PMID- 14738245 TI - Home mechanical ventilation in Hong Kong. AB - Home mechanical ventilation (HMV) is increasingly used to treat chronic respiratory failure. This present study was aimed to examine the trend, the disease categories treated and the outcomes of HMV use in Hong Kong. In the year 2002, all adult respiratory units in Hong Kong were invited to report to a multicentre retrospective survey of HMV use. A total of 249 patients (156 males, mean age 62.7 +/- 13.8 yrs) were treated since 1980, with 197 (79%) continuing with HMV at the time of the survey. Cumulative number of HMV grew as a cubic function of time since 1980. Currently, there are 2.9 users per 100,000 population. The predominant mode of HMV was noninvasive ventilation by bilevel pressure support ventilators (n=236). Chronic obstructive pulmonary disease accounted for 48.6% of all cases. The overall 3-yr HMV continuation rate was 66.2%. Death was the main reason for discontinuation. A rapidly rising trend of home mechanical ventilation use is observed in Hong Kong. In contrast to other series, chronic obstructive pulmonary disease was the major group treated and bilevel pressure support ventilation was the predominant mode chosen. Most patients tolerate home mechanical ventilation reasonably well, with approximately two-thirds continuing with its use at 36 months. PMID- 14738246 TI - KL-6 levels are elevated in plasma from patients with acute respiratory distress syndrome. AB - The acute respiratory distress syndrome (ARDS) is an extreme form of lung injury characterised by disruption to the alveolar epithelium. KL-6 is a mucin-like glycoprotein expressed on type II pneumocytes. Circulating levels of KL-6 have diagnostic and prognostic significance in a number of interstitial lung diseases, and when elevated are thought to indicate disruption of the alveolar epithelial lining. In this study, the authors sought to determine whether plasma KL-6 levels were elevated in patients with ARDS and whether these were associated with aetiology, disease severity, outcome or ventilatory strategy. Plasma samples were collected from 28 patients with ARDS, nine ventilated controls of matched illness severity and 10 healthy individuals. KL-6 concentrations were measured by enzyme linked immunosorbent assay. Patients with ARDS had higher plasma levels of KL-6 (median 537 U x mL(-1), interquartile range (IQR) 383-1,119), as compared to ventilated controls (median 255 U x mL(-1), IQR 83-338) and normal individuals (median 215 U x mL(-1), IQR 149-307). In patients with ARDS, plasma KL-6 levels were higher in nonsurvivors than survivors, and correlated positively with oxygenation index and negatively with arterial oxygen tension:inspiratory oxygen fraction ratio. There were also significant positive correlations with mean and peak airway pressures. Elevated levels of plasma KL-6 may provide a useful marker for acute respiratory distress syndrome in ventilated patients and have possible prognostic significance. Alveolar epithelial cell damage may be influenced by the nature of mechanical ventilatory support. PMID- 14738247 TI - New concepts of the pathogenesis of cystic fibrosis lung disease. AB - Although there has been impressive progress in the elucidation of the genetic and molecular basis of cystic fibrosis (CF), the pathogenesis of CF lung disease remains obscure. The elucidation of the pathogenesis of CF lung disease requires both a full description of normal innate airway defence and how absent function of the cystic fibrosis transmembrane regulator protein (CFTR) adversely perturbs this activity. Recent data have linked the abnormal ion transport properties of CF airway epithelia to depleted airway surface liquid (ASL) volume, reflecting the combined defects of accelerated Na+ transport and the failure to secrete Cl-. Depletion of a specific compartment of the ASL, i.e. the periciliary liquid (PCL), appears to abrogate both cilia-dependent and cough clearance. Subsequent to PCL depletion, mucus adheres to airway surfaces and persistent mucin secretion generates the formation of "thickened" mucus plaques and plugs, which become the nidus for bacterial infection. The paucity of liquid in these plaques/plugs, and the hypoxia in this environment, appear to promote biofilm bacterial infection. Therapeutic agents that restore airway surface liquid volume, i.e. blockers of Na+ transport, initiators of Cl- transport and osmolytes, are reviewed, as are strategies that may be required to use volume-restoring agents safely in patients with cystic fibrosis. PMID- 14738248 TI - Immunosuppressive therapy after human lung transplantation. AB - In 2002, equal numbers of lung transplantation (LTx) were performed with or without induction therapy with antilymphocyte antibodies, monoclonal anti-CD3 antibody or anti-interleukin-2-receptor monoclonal antibodies. It remains to be established if induction therapy after LTx is beneficial or deleterious for long term graft and patient survival. The vast majority of lung transplant recipients receive a triple-drug maintenance regimen including a calcineurin inhibitor, a cell-cycle inhibitor and steroids. Equal proportions receive cyclosporin A (CsA) and tacrolimus (Tac). There is also a trend to prescribe mycophenolate mofetil (MMF) instead of azathioprine (Aza). Steroid withdrawal is uncommon even 5 yrs after transplantation. The superiority of Tac over CsA as a maintenance agent has not been established to date, and the administration of MMF instead of Aza in combination with CsA and steroids did not improve graft or patient survival in a recent international, prospective, randomised, controlled trial. Shift from cyclosporin A to tacrolimus has emerged as the first treatment step of refractory acute rejection followed by high-dose steroids or antilymphocyte agents, total lymphoid irradiation or photopheresis. The treatment of chronic rejection remains deceptive and includes varied strategies such as modification of the maintenance regimen, addition of inhaled immunosuppressants and/or total lymphoid irradiation and photopheresis. PMID- 14738249 TI - Ciprofloxacin-induced acute interstitial pneumonitis. AB - The current authors present the case of a 68-yr-old female patient who developed severe respiratory failure after medication with ciprofloxacin for acute urinary tract infection. A chronic subdural haematoma was surgical evacuated. Postoperatively, an acute urinary tract infection was treated with ciprofloxacin. Six days later, C-reactive protein was rising and the patient was suffering from intermittent high fever, dyspnoea and severe hypoxaemia. The high-resolution computed tomography (HRCT) showed an interstitial lung disease in the anterior upper lobe on the left side as well as in the lingula. Assuming a bacterial infection amoxyl/clavulanic acid was started which did not improve the clinical symptoms. Bronchoalveolar lavage revealed a marked lymphocytosis (87%). Analysis for typical bacterial infections, Tuberculosis, Mycoplasma, Chlamydia and Legionella spp. were all negative. Another HRCT scan was made because of worsening of symptoms and this showed rapidly progressive infiltrates in most lobes. An open lingular biopsy showed an interstitial lymphoplasmocytotic infiltrate with some eosinophilic granulocytes and a few scattered giant cell granulomas, consistent with hypersensitivity pneumonitis. The patient's symptoms rapidly improved with systemic corticosteroid therapy and another HRCT scan revealed complete remission of pulmonary infiltrates. Ciprofloxacin can induce interstitial pneumonitis with acute respiratory failure. This is an important fact considering that ciprofloxacin is a widely used antibiotic agent in treatment of urinary tract infection. PMID- 14738250 TI - Fever, rigors and sweats in an immunocompromised male. PMID- 14738251 TI - Asthma: surfactant eliminates the early allergen-induced response. PMID- 14738252 TI - Role of interleukin-10 in idiopathic pulmonary fibrosis. PMID- 14738253 TI - Spirometry in young children. PMID- 14738254 TI - Injection safety in Pakistan: auto disable (AD) syringes in the curative sector. PMID- 14738255 TI - Socio-economic and demographic factors associated with injecting drug use among drug users in Karachi, Pakistan. AB - OBJECTIVE: To identify the socio-economic and demographic factors associated with injecting drug users (IDUs) in Karachi. METHODS: We recruited 242 IDUs (taking drugs through sub-dermal routes) and 231 non-IDUs (taking drugs other than sub dermal routes) from February through June 1996. IDUs were interviewed regarding sociodemographic factors, economic condition, and social network (marital status, living with spouse). In addition, information regarding location of drug users within the city (districts of Karachi) and current history of sexually transmitted diseases (STDs) were gathered. Moreover, blood samples were also obtained for HIV testing. RESULTS: Multivariate analysis showed that the income generation via illegal modes [AOR 1.6, 95% CI 1.0-2.6], non-sharing of income with family [AOR 1.7, 95% CI 1.1-2.7] and presence of suicidal thoughts [AOR 2.0, 95% CI 1.1-3.8] were associated with the use of drugs through injections. Further, drug users from districts West, East and Central were more likely to use drug through injection as compared to drug users from district South. The history of genital herpes was also found to be associated with injecting drug use. One IDU was found seropositive for HIV. CONCLUSION: The high-risk behaviors, such as illegal modes of earning and presence of suicidal thoughts, among IDUs suggest that the group needs rehabilitation programme. Moreover, non-sharing of income suggest that IDUs are isolated from social network, therefore primary prevention activities with focus on improving socio-economic conditions and social networking can reduce drug use through injections. Focused interventions on target districts would be helpful in reducing IDU. PMID- 14738256 TI - Frequency of Campylobacter jejuni in diarrhoea/dysentery in children in Rawalpindi and Islamabad. AB - OBJECTIVE: To determine the frequency of Campylobacter jejuni infection in children suffering from diarrhoea/dysentery in the Department of Microbiology, Army Medical College and Military Hospital, Rawalpindi, from 29 August 2002 to 29 November 2002. METHODS: The study was carried out on one hundred stool samples of children up to the age of twelve years admitted with diarrhoea/dysentery in Military hospital, Rawalpindi. The samples were collected in clean polypropylene containers containing Cary Blair medium. These were transported to the Microbiology Department, Army Medical College, Rawalpindi within 1-2 hours. The samples were inoculated on Modified Preston (Oxoid) and Karmali media (Oxoid) beside other routine stool culture media. The cultures were incubated at 42 degrees C under microaerophilic conditions. The growth after 48 hours was provisionally identified by colonial morphology, oxidase test, Gram staining and motility. The organisms were identified to species level by hippurate hydrolysis, urease test, nitrate reduction, catalase test, H2S production and resistance to cephalothin. RESULTS: Eighteen percent of samples yielded the growth of Campylobacter jejuni. Mean age of children with Campylobacter jejuni infection was 18 months with peak incidence from 12 to 21 months. Male female ratio was 1.7:1. All the children had loose motions. Seven out 18 (39%) had a combination of symptoms of loose motions, vomiting and pain abdomen. Those having fever with or without other complaints constituted 11 out of 18 (61.11%) i.e. more than 50% of all the children yielding C. jejuni had fever. About 90% of diarrhoeal stools had blood and fifty percent also had mucous. There was either history of chicken meat consumption or contact with cattle and pets in most of the cases and both in some of them. CONCLUSION: Campylobacter jejuni is a frequent cause of diarrhoea/dysentery in children in our set up. In children it is often related to pets keeping and chicken meat consumption. In the remaining, untreated drinking water may be the source. Campylobacter jejuni frequently presents with blood and mucous in stools with sporadic cases presenting with watery diarrhoea. PMID- 14738257 TI - Factors affecting unsuccessful referral by the Lady Health Workers in Karachi, Pakistan. AB - OBJECTIVE: To estimate the proportion of patient referral and to identify the factors associated with unsuccessful referral in Karachi, Pakistan. METHODS: In a cross-sectional survey, a total of 347 patients referred to different health care facilities by the LHWs were interviewed. Data was collected using a structured questionnaire, on demographic characteristics of patients (age, sex, income etc) and potential factors (seriousness of medical condition, distance of health facility, attitude of health center staff etc) that can affect patient referral outcome. RESULTS: An overall referral rate of 55% was found in this study. Out of a total of 347 patients interviewed, 265 (76.4%) were successful while 82 (23.6%) were unsuccessful referrals. Multivariate logistic regression analysis showed that objection to referral (Adjusted OR, 2.96; CI: 1.44-5.52), never referred before (Adjusted OR, 1.25; CI: 1.34-6.90), not visited the referral site before (Adjusted OR, 4.04; CI: 2.50-6.08) and no knowledge of who to meet at the referral site (Adjusted OR, 1.30; CI: 1.01-2.96) were the factors associated with unsuccessful referral. Other factors found significant were duration of the illness of the patient, anyone not known at the referral site and failure of LHW to follow up. CONCLUSION: Our study found that a significant proportion of patients seen by the LHWs are referred to different health facilities. Improved management skills of LHWs for simple medical problems would probably reduce the number of referrals. Efforts should also be directed to enhance the communication and counseling skills of LHWs, which may reduce a significant proportion of unsuccessful referrals. PMID- 14738258 TI - Blood lead levels during pregnancy and pregnancy outcome in Karachi women. AB - OBJECTIVE: To evaluate association of blood lead levels with pregnancy outcome in the obstetrics and gynaecology unit. METHODS: Blood lead levels were measured in 73 pregnant women at the time of delivery and assessed its association with pregnancy outcome. RESULTS: Mean maternal lead level was 9.91+/-4.44 mg/dL (range 2.28-36.35 mg/dL). Mothers of boys had significantly higher (p=0.013, one-tailed t test) blood lead levels (11.05+/-5.19) when compared to mothers of girls (8.74+/-3.18 mg/dL. CONCLUSION: Maternal lead levels at the time of delivery showed no association with gestational age, birth weight, recumbent length, or head circumference. PMID- 14738259 TI - Comparison of double disc and combined disc method for the detection of extended spectrum beta lactamases in enterobacteriaceae. AB - OBJECTIVE: To compare double disc approximation and combined disc method for their ability to detect extended spectrum b lactamase (ESBL) production in enterobacteriaceae and determine the percentage of isolates which are falsely reported as sensitive in absence of ESBL detection, in a clinical microbiology laboratory of a tertiary care hospital between September-October 2002. METHODS: Selected isolates were identified according to standard biochemical tests. Disc susceptibility tests were performed according to NCCLS. ESBL detection by combined disc [cefotaxime (30 ug) versus cefotaxime plus clavulanate (30+10 ug)] was compared with detection using double discs [amoxy-clavulanic acid (20+10 ug) and aztreonam (30 ug) applied 10 mm apart]. Results were interpreted according to NCCLS and analysed on SPSS version 10. RESULTS: ESBL production was detected in 140 (30%) isolates by combined disc method and 139 (29.5%) by double disc method. There was no significant difference between two methods. Of the ESBL positive isolates 41 (29%) gave zone diameters that were within the sensitivity range cutoff and would have been falsely reported as being beta lactam sensitive in absence of ESBL detection. CONCLUSION: ESBL detection should be routinely performed in clinical laboratories as false reporting would result in treatment failure despite in vitro sensitivity. No difference was found between the combined disc and double disc methods hence either of two could be used. PMID- 14738261 TI - Cutaneous manifestations of systemic lupus erythematosus in Pakistani patients. AB - OBJECTIVE: Systemic Lupus Erythematosus (SLE) is an autoimmune process in which cutaneous lesions occur in majority of patients. This study from Karachi, Pakistan was conducted to determine the pattern and prevalence of such lesions in SLE in Pakistani patients. METHODS: One hundred ninety eight patients with SLE fulfilling the clinical and laboratory criteria of the American Rheumatology Association were examined between 1986 and 2001 for the presence of cutaneous manifestations. RESULTS: Skin changes noted were: noncicatricial diffuse alopecia (22%), malar rash (31%), mucosal lesions (20%), discoid eruptions (15%), photosensitivity (33%), vascular lesions (20%), pruritus (17%), and pigmentary changes (22%). Peripheral gangrene, chronic ulcers, Raynauds phenomenon, urticaria, chilblains, thrombophlebitis, palmar erythema, and erythema multiform were rare. Anti ANA and anti dsDNA were positive in 93% and 83% patients respectively. CONCLUSION: A different clinical pattern was noted in our patients than reported previously. PMID- 14738260 TI - Use of intravenous anti-D in patients with refractory and relapsed immune thrombocytopenic purpura. AB - OBJECTIVE: To determine the response to i.v. anti-D and its comparison with splenectomy as second line therapy in refractory and relapsed cases of ITP, in the Aga Khan University Hospital, Karachi. METHODS: A total of 23 patients with chronic ITP were treated with either anti-D or splenectomy as second line treatment. The patients were assessed for time to achieve a response to second line treatment, duration of response and adverse events. RESULTS: There were 12 patients in the anti-D group and 11 in the splenectomy group. The mean platelet count at presentation was 9,000/cumm. The mean age was 8.9 years and 13.0 years and the male to female ratio was 1:1 and 1:1.2 in anti-D and splenectomy group respectively. 54.5% of the patient in the anti-D group responded compared to 81.8% in the splenectomy group. Median time to achieve a response was 7 days in the anti-D group and 1 day in the splenectomy group. Mean time to relapse was 87.8 days in the anti-D group and 55.4 days in the splenectomy group. No adverse events were recorded for any of the infusions of anti-D and none of the patients had more than 0.5 gm/dl fall in the hemoglobin level following anti-D infusion. CONCLUSION: It was thus concluded that Anti-D is a relatively safe, convenient and effective therapy for chronic ITP and can be used as a splenectomy sparing agent when treatment is clinically indicated. PMID- 14738262 TI - Midazolam and thiopentone co-induction: looking for improvement in quality of anaesthesia. AB - OBJECTIVE: To evaluate improvement in quality of anaesthesia induction using thiopentone and midazolam for co-induction of anaesthesia. An additional end point was taken as loss of response to a tetanic stimulation (50 Hz) delivered for 5 seconds after the loss of verbal contact and eyelash reflex. METHODS: Ninety ASA I and II patients, within the age range of 20-60 years were studied. Patients were randomly divided into three equal groups; A, B and group C. RESULTS: Onset of induction and loss of response to a tetanic stimulation was achieved earlier in group B as compared to the other study groups (p<0.05). CONCLUSION: Co-induction with midazolam 0.02 mg x kg(-1) followed by thiopentone 3 mg x kg(-1) was superior to other two groups. Induction of anaesthesia in this group was found to be smoother and faster, provided better hemodynamic stability, better airway maintenance and with lesser incidence of untoward effects. PMID- 14738263 TI - Impact of Gemcitabine and Cisplatin with radiotherapy in locally advanced or metastatic transitional cell carcinoma of urinary bladder. AB - OBJECTIVE: The primary object of this study is to evaluate the efficacy and safety of Gemcitabine and Cisplatin along with radiotherapy in transitional cell carcinoma of urinary bladder. PATIENTS AND METHODS: Twenty patients with locally advanced or metastatic TCC of urinary bladder were enrolled during the 22-months period from January, 1999 to October, 2000 and followed up till March 2002. Three patients received 4 cycles, five patients received 5 cycles and twelve patients received 6 cycles of Gemcitabine 1250 mg/m2 on day 1 and day 8 and Cisplatin 80 mg/m2 on day 1; administered every 3 weeks. No patient received prior chemotherapy, radiotherapy or surgery. However, four patients received prior intravesical chemotherapy. All patients received radiotherapy after completion of chemotherapy regimen. RESULTS: Nineteen patients achieved complete response at the end of the treatment. The complete response rate was 95%. The confidence interval was at 95%, level of confidence ranged from 85% to 100%. Median duration of clinical benefit was 21 months. Six patients (30%) were documented neutropenia, three patients (15%) documented thrombocytopenia. No life threatening toxicity was observed. CONCLUSION: Gemcitabine and Cisplatin along with radiotherapy in locally or metastatic Transitional cell carcinoma of urinary bladder, exhibited pronounced response rate among all the patients. The toxicity profile remained extremely low and disease free survival enhanced. The above investigation may further be continued at a larger scale encompassing a wide band of subjects. PMID- 14738264 TI - Cost of acute stroke care at a tertiary care hospital in Karachi, Pakistan. AB - OBJECTIVE: To evaluate cost of acute stroke care and its determinants at a tertiary care hospital in Karachi and to find out predictors of high cost care. Acute stroke is a leading cause of morbidity and mortality. Cost of care is the single most important determinant in availability of acute stroke care at a tertiary care hospital in Pakistan. It is also an important factor in development of public health policies and medical insurance plans. Average annual income in Pakistan is 4881 rupees (85 U dollars). METHODS: Medical and billing records of 443 patients with acute stroke were retrospectively reviewed from 1998-2001 at The Aga Khan University Hospital (AKUH), Karachi. Acute stroke care at AKUH usually includes routine laboratory investigation including Lipid profile, Magnetic resonance imaging/angiography (MRI/MRA), Echocardiogram, Carotid Doppler's ultrasound and medical management in the Stroke care unit. RESULTS: 443 patients were included in study. Age range was 25-98 years (Mean 58 years). 269 (61%) were male. Length of hospital stay was 1 day; 67 patients, 2 days; 83 patients, 3 days; 70 patients, 4-5 days; 87 patients, 6-10 days; 75 patients, 11 30 days; 49 patients and more than 30 days; 12 patients. Average length of stay was five days and median length was three days. Average total cost was 70,714 rupees (1179 U dollars) which included average radiology cost; 12,507 rupees (208 U dollars), average laboratory cost; 8365 rupees (139 U dollars), average pharmacy cost; 13,320 rupees (222 U dollars) and average bed/room charges; 27,552 rupees (459 U dollars). Length of hospital stay is the most important determinant of cost. Average total cost for patients who stayed for 1 day was 19,597 rupees (326 U dollars), 2-3 days; 25,568 rupees (426 U dollars), 4-7 days; 49,705 rupees (828 U dollars), 8-30 days; 153,586 rupees (2559 U dollars), more than 30 days; 588,239 rupees (9804 U dollars). Average cost for general ward was 60,574 rupees (1010 U dollars), private ward was 74,880 rupees (1248 U dollars) and intensive care unit was 155,010 rupees (2583 U dollars). CONCLUSION: Cost of acute stroke care is extremely high as compared to average national income at our hospital. Most important determinant of cost is length of hospital stay. Cost cutting measures and increased funding from state are necessary to increase the availability of acute stroke care. PMID- 14738265 TI - Food insecurity in metropolis of the developing world--observations from central district of Karachi, Pakistan. AB - OBJECTIVE: To assess the prevalence of household food insecurity at various income levels in urban areas of Karachi. METHODS: A cross sectional survey of different localities in the central district of Karachi was carried out. The data was collected through household interviews of housewives conducted by trained nutritionist. Housewives were interviewed about food security status of the household. RESULTS: A total of 797 families were visited from the central district of Karachi. Mean Food Security Score increased with income level. The difference was statistically significant (ANOVA) between 1st and second (P=0.000) and 2nd and 3rd group (P=0.000) but not between 3rd and fourth group. At the very low and low income levels 83% & 51% families respectively were food insecure in any degree, while this percentage was very low at the middle (6.3%) and high income level (1.8%). The difference in prevalence of food insecurity between the VLI and LI, and, LI and MI income groups was statistically significant (chi.sq. test, p<0.001 in each case). Hunger (because of lack of money) was experienced in the preceding year only by "very low income" (37%) and "low income" families (17%). Use of coping strategies was most frequently mentioned for parents and then for children and only occasionally for infants or grand parents. Females were more likely to be effected by food insecurity than males. Meat, milk and fruits were the food groups which were preferred and considered healthy but were avoided because of lack of money by a majority (51%-86%) of families. CONCLUSIONS: In spite of having an impression of being an affluent city of Pakistan, the prevalence of food insecurity with and without hunger is rampant not only among very low income (slum dwellers) but also among low income families of Karachi. PMID- 14738266 TI - Genetic aspects of human obesity: a review. PMID- 14738267 TI - Resolution of intracranial tuberculoma with medical therapy. PMID- 14738268 TI - Blue rubber bleb nevus syndrome: associated with severe GI bleeding requiring one hundred blood transfusions. PMID- 14738269 TI - Giant choroid plexus papilloma of the third ventricle. PMID- 14738270 TI - Environmental contamination and health studies: conflicts of interest and reasons for community-based participatory studies. PMID- 14738271 TI - Toxicology of metals--science confused by poor use of terminology. PMID- 14738272 TI - Cancers among residents downwind of the Hanford, Washington, plutonium production site. AB - A community-based health survey for the time period between 1944 and 1995 was collected from 801 individuals who had lived downwind of the U.S. plutonium production facility located in Hanford, Washington. The results of the survey revealed high incidences of all cancers, including thyroid cancer. There were greater than expected numbers of central nervous system tumors and cancers that invaded the female reproductive system (e.g., cancers of the uterus, ovary, cervix, and breast). The authors argue that the greater-than-expected numbers found cannot be accounted for by selection bias alone. Comparisons of crude incidence rates, as well as of occurrence ratios between pairs of cancer types among Downwinders and reasonably similar populations, suggested that the excess neoplasms may be associated with radioactive contamination of food, water, soil, and/or air. In addition, a synergistic effect may exist with agricultural toxins. Previously neglected biophysical and physiological properties of internally lodged, long-lived 129I may be a significant etiological factor in the development of thyroid diseases, including cancer, and other malignancies in exposed populations. PMID- 14738273 TI - Microbial exposure and mold-specific serum IgG levels among children with respiratory symptoms in 2 school buildings. AB - In this study, the authors determined the association between serum mold-specific immunoglobulin G (IgG) levels in primary school children (N = 181) with asthma, wheezing, or cough symptoms and exposure to indoor mold in 2 schools, with and without mold damage. Microbial exposure was determined with environmental sampling. Serum IgG antibody concentrations against 24 microbial strains were determined with enzyme-linked immunosorbent assay. Children's microbe-specific IgG levels were often higher in the reference school. There was an association between elevated serum IgG antibody levels to Penicillium notatum and moisture damage in the school. In addition, moisture damage both in school and at home was associated with Penicillium notatum and Eurotium amstelodami IgG levels. These observations comported with microbial findings in the index school. No other positive associations between IgG antibodies and microbial findings for the school buildings were observed. IgG levels in sera of school children did not provide a specific means for assessing the microbial exposure associated with moisture and microbial problems in the school buildings. Thus, IgG analysis cannot be readily suggested as a routine method for the evaluation of these exposures. PMID- 14738274 TI - Case series: use of induced sputum in the evaluation of occupational lung diseases. AB - The authors recently reported that analysis of induced sputum may reveal the status of hazardous dust exposure (e.g., silica, hard metals) as effectively as does bronchoalveolar lavage. In the current study, the authors describe how induced sputum can assist in the evaluation and diagnosis of suspected occupational lung diseases. The 3 patients who underwent induced sputum testing included a miner with silicosis, a dental technician with berylliosis, and a teacher who suffered from undefined interstitial fibrosis and in whom induced sputum analysis revealed the presence of a high burden of calcium sulfate and silica. The data reported indicate that induced sputum--a known safe and simple procedure--can serve as a useful tool in the evaluation of patients with suspected occupational lung diseases. PMID- 14738275 TI - Ventilatory function of progressive massive fibrosis among bituminous coal miners in Taiwan. AB - Geographic and ethnic differences exist for the effects of respirable coal-mine dust on the lung function of miners. In this study, the authors compared 177 coal workers who had radiological evidence of progressive massive fibrosis (PMF) with 87 healthy male control subjects. The authors performed maximal expiratory flow volume measurements, single-breath carbon monoxide diffusing capacity (DLco) measurements, and arterial blood gas analysis on each subject. The data revealed that miners with early PMF (category A) had significantly reduced, but well preserved, vital capacity (VC) and forced expiratory volume in 1 sec (FEV1.0), whereas FEV1.0/VC and DLco were decreased in both nonsmokers and smokers. Abnormally low (i.e., < 80% of predicted values) VC and FEV1.0, and further decreases in DLco, were observed in miners with late PMF (categories B and C). The predominant impairment patterns for workers in categories A, B, and C were obstructive, obstructive and mixed, and mixed and restrictive, respectively. Smoking increased the magnitude of airway obstruction. The authors concluded that diversity in functional impairment was present among bituminous coal miners, even among those with PMF. PMID- 14738276 TI - Polycyclic aromatic hydrocarbon-deoxyribonucleic acid (PAH-DNA) adduct levels and exposure to coke oven emissions among workers in Taiwan. AB - In this study, the authors evaluated the relationship between polycyclic aromatic hydrocarbon-deoxyribonucleic acid (PAH-DNA) adduct levels in workers' peripheral white blood cells (WBCs) and their occupational exposures to coke oven emissions (measured by air and urinary 1-hydroxypyrene [1-OHP]). Personal exposures to smoking, charbroiled food, changing clothes in the workplace, and respirator use were determined by questionnaire. Eighty-nine coke oven workers were divided into 3 exposure groups on the basis of job description: topside workers, cokeside workers, and plant office staff. Referent subjects comprised 63 individuals from the same company who worked at a site remote from the coking plant. The geometric mean (GM) PAH-DNA adduct levels determined from the WBCs of the exposed groups were 6.86, 1.56, and 0.90 adducts/10(8) nucleotides, respectively (referents = 0.38 adducts/10(8) nucleotides). GM personal benzene soluble fraction (BSF) exposures for the exposed groups were 483.2 microg/m3, 70.1 microg/m3, and 43.2 microg/m3, respectively (referents = 10.7 microg/m3). There was a significant correlation (p < 0.05) between individual BSF and PAH-DNA adduct levels for the exposed groups. The authors also found a significant correlation (p < 0.05) between urinary 1-OHP levels from the day 2 samples of the exposed groups and their PAH-DNA adduct levels. The logistic-regression model revealed that PAH-DNA adduct levels were significantly different between job categories. The results of this study indicate that BSF exposure is the primary contributor to PAH-DNA adduct levels determined from WBCs. PMID- 14738277 TI - Serum fluoride levels in a group of Egyptian infants and children from Cairo city. AB - In this study, the authors investigated fluoride levels in the serum of infants and children (n = 296) and in the breast milk from nursing mothers (n = 60) in Cairo city. Their goal was to evaluate the necessity and safety of implementing a fluoride supplementation program. The authors used an ion-selective electrode to assay fluoride by direct potentiometry. Also, 2- to 12-yr-old participants underwent clinical dental examinations to detect caries and/or fluorosis. The serum fluoride levels of infants were significantly lower than levels found in preschoolers and school-age children. Serum fluoride correlated positively with age; it was significantly lower during the 1st than 2nd yr of infancy (p = 0.005). Breast or formula feeding did not influence serum fluoride status; the fluoride levels in mothers' milk reflected the serum levels of their own infants. Dental examinations revealed that 81% of the children had caries, whereas there was no evidence of fluorosis. Serum fluoride levels did not vary with the presence or absence of dental caries and did not correlate with the number of decayed, missing, or filled teeth. Gender did not influence serum fluoride expression, and the percentile values were unrelated to height, weight, or head circumference. These findings suggest the necessity and safety of improving the fluoride consumption levels of infants and children in Cairo city. Wider-scale studies are needed to obtain better insight into the problem. PMID- 14738278 TI - There is an alternative reason for lower-than-expected rates of lung cancer in farmers. PMID- 14738279 TI - Neuropsychological effects of treatments for adults with cancer: a meta-analysis and review of the literature. AB - A meta-analysis was conducted to evaluate possible neuropsychological effects of treatments for cancer in adults. A search revealed 30 studies, encompassing 29 eligible samples, and leading to inclusion of a total of 838 patients and control participants. A total of 173 effect sizes (Cohen's d) were extracted across 7 cognitive domains and as assessed in the literature via 3 methods of comparison (post-treatment compared with normative data, controls, or baseline performance). Statistically significant negative effect sizes were found consistently across both normative and control methods of comparison for executive function, verbal memory, and motor function. The largest effects were for executive function and verbal memory normative comparisons (-.93 and -.91, respectively). When limiting the sample of studies in the analyses to only those with relatively "less severe" diagnoses and treatments, the effects remained. While these results point toward some specific cognitive effects of systemic cancer therapies in general, no clear clinical implications can yet be drawn from these results. More research is needed to clarify which treatments may produce cognitive decrements, the size of those effects, and their duration, while ruling out a wide variety of possible mediating or moderating variables. PMID- 14738280 TI - Caloric stimulation in neglect: evaluation of response as a function of neglect type. AB - Contralesional neglect may be induced by either unawareness of contralesional stimuli (attentional neglect, AN) or failure to act in contralesional space (intentional neglect, IN). We examined whether contralesional cold caloric stimulation differentially affects AN versus IN. Patients with left-sided neglect (n = 16) from right-hemisphere lesions performed target cancellation and line bisection tasks. Using a video-based apparatus that reverses the right and left side of stimuli, patients with abnormal cancellation performance were divided into those with AN and those with IN. The 5 subjects with normal cancellation performance but rightward bisection bias were also separated into 2 neglect groups. Subjects performed cancellation or bisection tasks before and immediately after irrigation of the left auditory canal with ice water. Caloric stimulation induced brisk rightward nystagmus in all subjects. Subjects with AN cancelled more left-sided targets after stimulation than those with IN (p = .02). Whereas caloric stimulation significantly shifted bisection error leftward for both IN and AN groups (p < .0001), AN patients exhibited a greater magnitude of shift than the IN patients. While the basis for differential performance remains undefined, the data indicate that caloric stimulation influences AN more than IN. PMID- 14738281 TI - Intra-individual measures of association in neuropsychology: inferential methods for comparing a single case with a control or normative sample. AB - Performance on some neuropsychological tests is best expressed as an intra individual measure of association (such as a parametric or non-parametric correlation coefficient or the slope of a regression line). Examples of the use of intra-individual measures of association (IIMAs) include the quantification of performance on tests designed to assess temporal order memory or the accuracy of time estimation. The present paper presents methods for comparing a patient's performance with a control or normative sample when performance is expressed as an IIMA. The methods test if there is a significant difference between a patient's IIMA and those obtained from controls, yield an estimate of the abnormality of the patient's IIMA, and provide confidence limits on the level of abnormality. The methods can be used with normative or control samples of any size and will therefore be of particular relevance to single-case researchers. A method for comparing the difference between a patient's scores on two measures with the differences observed in controls is also described (one or both measures can be IIMAs). All the methods require only summary statistics (rather than the raw data from the normative or control sample); it is hoped that this feature will encourage the development of norms for tasks that use IIMAs to quantify performance. Worked examples of the statistical methods are provided using data from a clinical case and controls. A computer program (for PCs) that implements the methods is described and made available. PMID- 14738282 TI - Predictors of postconcussion symptom complex in community dwelling male veterans. AB - The presence of a persistent postconcussion symptom complex (PPCSC) was examined in a non-referred sample of male veterans with a history of mild head injury and a comparison group without a history of head injury. Hierarchical logistic regression procedures were used to determine possible predictors of PPCSC using variables supported by previous research (i.e., preexisting psychiatric difficulties, demographic and social support variables, and history of an accidental injurious event). Although PPCSC was common in all groups (23% of the total sample), a significantly greater proportion of individuals in the mild head injury with loss of consciousness group (37.2%) had PPCSC compared with three other groups (head injury without loss of consciousness = 26.1%; motor vehicle accident without head injury = 23%; and control = 17.3%). However, the most salient predictors of PPCSC were early life psychiatric difficulties such as anxiety or depression, limited social support, lower intelligence, and interactions among these variables. The predictive value of loss of consciousness was significant, but low (1.4% of unique variance). The findings provide support for the premise that PPCSC is mediated in part by individual resilience, preexisting psychological status, and psychosocial support. PMID- 14738283 TI - Learning in children and sleep disordered breathing: findings of the Tucson Children's Assessment of Sleep Apnea (tuCASA) prospective cohort study. AB - We examined the relationship between nocturnal respiratory disturbance and learning and compared learning in children with and without nocturnal respiratory disturbance. Subjects were 149 participants in a prospective cohort study examining sleep in children ages 6-12: The Tucson Children's Assessment of Sleep Apnea study (TuCASA). Sleep was assessed via home polysomnography. Intelligence, learning and memory, and academic achievement were assessed. Parents rated attention. Group comparisons were used to test the hypothesis that the group with an apnea/hypopnea index (AHI) of 5 or more (n = 77) would have weaker performance than the group with AHI less than 5 (n = 72). The group with AHI of 5 or more had weaker learning and memory though differences between groups decreased when arousals were taken into account. There was a greater percentage of Stage 1 sleep in the AHI 5 or more group, and Stage 1 percentage was negatively related to learning and memory in the sample (n = 149). There were negative relationships between AHI and immediate recall, Full Scale IQ, Performance IQ, and math achievement. Hypoxemia was associated with lower Performance IQ. Thus, findings suggest that nocturnal respiratory disturbance is associated with decreased learning in otherwise healthy children, that sleep fragmentation adversely impacts learning and memory, and that hypoxemia adversely influences nonverbal skills. PMID- 14738284 TI - Neural hybrid model of semantic object memory: implications from event-related timing using fMRI. AB - Previous studies by our group have demonstrated fMRI signal changes and synchronized gamma rhythm EEG oscillations between thalamus and cortical regions as subjects recall objects from visually presented features. Here, we extend this work by estimating the time course of fMRI signal changes in the cortical and subcortical regions found to exhibit evidence for task-related activation. Our results indicate that there are separate loci of signal changes in the thalamus (dorsomedial and pulvinar) that exhibit notable differences in times of onset, peak and return to baseline of signal changes. The signal changes in the pulvinar demonstrate the slowest transients of all the cortical and subcortical regions we examined. Evaluation of cortical regions demonstrated salient differences as well, with the signal changes in Brodmann area 6 (BA6) rising, peaking, and returning to baseline earlier than those detected in other regions. We conclude that BA6 mediates early designation or refinement of search criteria, and that the pulvinar may be involved in the binding of feature stimuli for an integrated object memory. PMID- 14738285 TI - The basal ganglia and semantic engagement: potential insights from semantic priming in individuals with subcortical vascular lesions, Parkinson's disease, and cortical lesions. AB - The impact of basal ganglia dysfunction on semantic processing was investigated by comparing the performance of individuals with nonthalamic subcortical (NS) vascular lesions, Parkinson's disease (PD), cortical lesions, and matched controls on a semantic priming task. Unequibiased lexical ambiguity primes were used in auditory prime-target pairs comprising 4 critical conditions; dominant related (e.g., bank-money), subordinate related (e.g., bank-river), dominant unrelated (e.g., foot-money) and subordinate unrelated (e.g., bat-river). Participants made speeded lexical decisions (word/nonword) on targets using a go no-go response. When a short prime-target interstimulus interval (ISI) of 200 ms was employed, all groups demonstrated priming for dominant and subordinate conditions, indicating nonselective meaning facilitation and intact automatic lexical processing. Differences emerged at the long ISI (1250 ms), where control and cortical lesion participants evidenced selective facilitation of the dominant meaning, whereas NS and PD groups demonstrated a protracted period of nonselective meaning facilitation. This finding suggests a circumscribed deficit in the selective attentional engagement of the semantic network on the basis of meaning frequency, possibly implicating a disturbance of frontal-subcortical systems influencing inhibitory semantic mechanisms. PMID- 14738286 TI - Syntactic language processing: ERP lesion data on the role of the basal ganglia. AB - The role of the basal ganglia in syntactic language processing was investigated with event-related brain potentials in fourteen neurologically impaired patients. Seven of these patients had basal ganglia lesions while 7 other patients primarily had lesions of the left temporo-parietal region excluding the basal ganglia. All patients listened to sentences that were either correct or included a verb argument structure violation. In previous experiments this type of violation elicited a biphasic pattern of an N400-P600 complex in young healthy participants. While the N400 may result from incorrect semantic-thematic role assignment, the P600 reflects the fact that verb information does not license the syntactic structure at present. Results of the patient experiment revealed a double dissociation: patients with left temporo-parietal lesions only show a P600, whereas patients with lesions of the basal ganglia showed no P600, but a negativity with extended duration that resembled an N400. The latter pattern not only confirms previous reports that the basal ganglia modulate the P600 but extends these results by showing that the N400 as a late semantic-thematic integration process appears partially modulated by the basal ganglia. PMID- 14738287 TI - Left and right basal ganglia and frontal activity during language generation: contributions to lexical, semantic, and phonological processes. AB - fMRI was used to determine the frontal, basal ganglia, and thalamic structures engaged by three facets of language generation: lexical status of generated items, the use of semantic vs. phonological information during language generation, and rate of generation. During fMRI, 21 neurologically normal subjects performed four tasks: generation of nonsense syllables given beginning and ending consonant blends, generation of words given a rhyming word, generation of words given a semantic category at a fast rate (matched to the rate of nonsense syllable generation), and generation of words given a semantic category at a slow rate (matched to the rate of generating of rhyming words). Components of a left pre-SMA-dorsal caudate nucleus-ventral anterior thalamic loop were active during word generation from rhyming or category cues but not during nonsense syllable generation. Findings indicate that this loop is involved in retrieving words from pre-existing lexical stores. Relatively diffuse activity in the right basal ganglia (caudate nucleus and putamen) also was found during word generation tasks but not during nonsense syllable generation. Given the relative absence of right frontal activity during the word generation tasks, we suggest that the right basal ganglia activity serves to suppress right frontal activity, preventing right frontal structures from interfering with language production. Current findings establish roles for the left and the right basal ganglia in word generation. Hypotheses are discussed for future research to help refine our understanding of basal ganglia functions in language generation. PMID- 14738288 TI - Mental object rotation in Parkinson's disease. AB - Deficits in visual-spatial ability can be associated with Parkinson's disease (PD), and there are several possible reasons for these deficits. Dysfunction in frontal-striatal and/or frontal-parietal systems, associated with dopamine deficiency, might disrupt cognitive processes either supporting (e.g., working memory) or subserving visual-spatial computations. The goal of this study was to assess visual-spatial orientation ability in individuals with PD using the Mental Rotations Test (MRT), along with other measures of cognitive function. Non demented men with PD were significantly less accurate on this test than matched control men. In contrast, women with PD performed similarly to matched control women, but both groups of women did not perform much better than chance. Further, mental rotation accuracy in men correlated with their executive skills involving mental processing and psychomotor speed. In women with PD, however, mental rotation accuracy correlated negatively with verbal memory, indicating that higher mental rotation performance was associated with lower ability in verbal memory. These results indicate that PD is associated with visual-spatial orientation deficits in men. Women with PD and control women both performed poorly on the MRT, possibly reflecting a floor effect. Although men and women with PD appear to engage different cognitive processes in this task, the reason for the sex difference remains to be elucidated. PMID- 14738289 TI - Neural basis for impaired time reproduction in Parkinson's disease: an fMRI study. AB - Studies involving brain-lesioned subjects have used the paced finger tapping (PFT) task to investigate the neural systems that govern motor timing. Patients with Parkinson's disease (PD), for example, demonstrate abnormal performance on the PFT, characterized by decreased accuracy and variability changes, suggesting that the basal ganglia may play a critical role in motor timing. Consistent with this hypothesis, an fMRI study of healthy participants demonstrated that the medial frontostriatal circuit (dorsal putamen, ventrolateral thalamus, SMA) correlated with explicit time-dependent components of the PFT task. In the current fMRI study, PD patients and healthy age-matched controls were imaged while performing the PFT. PD patients underwent 2 imaging sessions, 1 on and the other off dopamine supplementation. Relative to controls, PD patients were less accurate and showed greater variability on the PFT task relative to controls. No PFT performance differences were observed between the on and off medication states despite significantly greater motor symptoms on the Unified Parkinson's Disease Rating Scale (UPDRS) in the off medication state. Functional imaging results demonstrated decreased activation within the sensorimotor cortex (SMC), cerebellum, and medial premotor system in the PD patients compared to controls. With dopamine replacement, an increase in the spatial extent of activation was observed within the SMC, SMA, and putamen in the PD patients. These results indicate that impaired timing reproduction in PD patients is associated with reduced brain activation within motor and medial premotor circuits. Despite a lack of improvement in PFT performance, PD patient's brain activation patterns were partially "normalized" with dopamine supplementation. These findings could not be attributed to greater head movement artifacts or basal ganglia atrophy within the PD group. PMID- 14738290 TI - Dual monoclonal antibody therapy for the treatment of PTLD? PMID- 14738291 TI - Preventing acute and chronic rejection after diagnosis of post-transplantation lymphoproliferative disorder. PMID- 14738292 TI - Is decreased bone mineral density in pediatric transplant recipients really a problem? PMID- 14738293 TI - The potential impact of SARS on organ transplantation: exercise caution. PMID- 14738294 TI - Facilitating cells as a venue to establish mixed chimerism and tolerance. AB - Graft rejection and the toxicity associated with the use of non-specific immunosuppression remain the major limitations in pediatric solid organ transplantation. The induction of tolerance in transplant recipients is an elusive but achievable goal that will decrease the dependence on immunosuppressive agents. BMT is associated with a robust form of donor-specific transplantation tolerance. It achieves a state of chimerism, defined as the presence of donor marrow cells in the recipient. The two major toxicities in conventional bone marrow transplantation that have prevented its clinical application to induce tolerance are the toxicity of ablative conditioning and GVHD. Two forms of chimerism exist: full chimerism and mixed chimerism. In full chimerism, the hematopoietic system of the recipient is replaced by that of the donor following ablative conditioning. Full chimerism is associated with a relatively impaired immunocompetence for primary immune responses and an increased risk of GVHD. In addition, the 7-10% regimen-related mortality associated with ablation could not be accepted in solid organ allograft recipients. In mixed chimerism the donor hematopoietic system co-exists with that of the recipient. Mixed chimerism induces donor-specific tolerance and is associated with superior immunocompetence and a relative resistance to GVHD compared with full chimerism. Moreover, it can be achieved with partial conditioning, thereby reducing the regimen-related morbidity associated with myeloablation. Approaches to establish mixed chimerism using non-myeloablative conditioning regimens have been aggressively pursued over the past decade. Mixed chimerism can be safely established with minimal conditioning, resulting in a significant reduction in risk compared with ablative conditioning. GVHD is the final hurdle that has prevented the widespread application of chimerism to induce tolerance. Donor T cells are the primary effector cells for GVHD. Although T cell depletion of the donor marrow avoids GVHD, it results in an increase in the rate of graft failure in MHC-disparate recipients. The dichotomy between GVHD and T cell depletion graft failure has recently been dissociated by the discovery of CD8+/TCR- graft FC. Purified HSC engraft readily in syngeneic recipients but not in MHC-disparate allogeneic recipients. The addition of small numbers of facilitating cells permits durable HSC engraftment in allogeneic recipients and avoids GVHD. Using FC to promote HSC engraftment following non-myeloablative conditioning could be a promising approach to establish tolerance in solid organ transplantation. This invited review focuses on recent developments in stem cell chimerism and tolerance that could bring the use of this approach to induce tolerance to solid organ transplantation one step closer to reality. PMID- 14738295 TI - Decreased bone mineral density in the pediatric renal transplant population. AB - All renal transplant recipients at our centre have had bone mineral density assessment (BMD) by DEXA scans of their lumbar spine while on the transplant waitlist and at 6-month intervals post-transplant over the past 7 yr. Risk factors for osteopenia and osteoporosis including donor source, dialysis status prior to transplantation, prior renal disease, and biopsy confirmed rejection events and their relationship to BMD of the lumbar spine were assessed. Thirty nine children transplanted over the past 7 yr were included in this study. In total, 127 BMD longitudinal assessments were performed. From 1990 to 1997, ATG/ALG was used as antibody induction therapy. From 1997 to 2002, Basiliximab was utilized. Cyclosporin A (CyA) was the primary immunosuppressant for most children with tacrolimus as primary (n = 2) and switch for CyA failure or toxicity (n = 16). Prednisone was administered at a dose of 1 mg/kg/day for the first week and tapered to 10 mg/m2/alternate day by 1 month post-transplant. Azathioprine 1.5 mg/kg/day was continued for 1 yr and discontinued in children who were rejection free. All rejections were biopsy confirmed and treated with a prednisone pulse. Using a repeated measures regression analysis, we have found that L1-L4 BMD z score is affected by height and transplant number. It is also related to time relative to transplant in a quadratic fashion. There was an inverse relationship between advancing patient age and L1-L4 BMD z score. L1-L4 BMD z score was not related to weight, pre-existing renal disease, gender, donor source, type of renal replacement therapy prior to transplantation, or rejection events. PMID- 14738296 TI - Survival of childhood polycystic kidney disease following renal transplantation: the impact of advanced hepatobiliary disease. AB - Childhood PKD encompasses the diagnoses of AR and ADPKD, glomerulocystic disease, and syndromes such as tuberous sclerosis or Jeune's syndrome. Given the fact that a majority of PKD children with ESRD carry the diagnosis of ARPKD, natural history studies assessing the long-term prognosis of PKD patients following renal transplantation must focus on morbidity and mortality issues related to complications from congenital hepatic fibrosis. Using the NAPRTCS registry, we analyzed the patient and graft survival rates of 203 PKD patients and 7044 non PKD patients undergoing renal transplantation between 1987 and 2001. Deceased PKD patients, all with a diagnosis of ARPKD, were further identified and characterized using a special questionnaire submitted to the principal investigators. Overall graft and patient survival rates were not significantly different between PKD and non-PKD patients. No differences in rates of acute rejection or time to first rejection were noted between PKD and non-PKD patients. The relative risk of living longer than 3 yr in the PKD patients was not significantly different from non-PKD patients (RR = 0.70, p = 0.28). Sepsis was identified as a likely factor in the cause of death in nine (64%) ARPKD patients and was comfirmed with a positive blood culture in four patients. Despite similar graft and patient survival rates among PKD and non-PKD children following renal transplantation, our results suggest that ARPKD transplant recipients appear to be at increased risk for sepsis that may be related to hepatic fibrosis and ascending cholangitis. The utility of early liver transplantation in ARPKD patients with significant hepatobiliary disease is discussed. PMID- 14738297 TI - Lymphocyte subsets may discern treatment effects in children and young adults with post-transplant lymphoproliferative disorder. AB - To identify potential alternatives to Epstein-Barr virus (EBV)-specific cytotoxic T-cell responses, peripheral lymphocyte subsets (PLS) (CD4+, CD8+, CD3+, CD19+, CD56+) were measured by flow cytometry in children with abdominal transplants (n = 22) and heart transplants (n = 2), with (n = 14) and without (n = 10, group C) post-transplant lymphoproliferative disorder (PTLD). PTLD resolved with reduced immunosuppression and antiviral therapy in eight children (group B). Recalcitrant PTLD was observed in six children (group A). Recalcitrant PTLD followed prior antilymphocyte therapy [monoclonal anti-CD3 antibody (OKT3) and thymoglobin (n = 3) and thymoglobin (n = 1)] for refractory rejection in four of these six children, and resolved after treatment with rituximab (anti-CD20 monoclonal antibody). Ten children without PTLD served as a control group (group C). Between group comparisons showed a numeric increase in CD8 + cells and significantly lower CD4:CD8 ratios in both PTLD groups (A and B) compared with group C. Group A children also demonstrated significant depletion of natural killer (NK) cells, and post-rituximab depletion of B-cells compared with group B (no rituximab treatment). We conclude that NK cell depletion with a reversed CD4:CD8 ratio may represent a persistent immunosuppressed state, which may result from prior antilymphocyte therapy and may predispose to recalcitrant EBV-PTLD. Clinical remission with rituximab is accompanied by B-cell depletion. Serial monitoring of PLS from the time of diagnosis of PTLD will be necessary to confirm these observations. PMID- 14738298 TI - Use of prophylactic lamivudine and mycophenolate mofetil in renal transplant recipients with chronic hepatitis B infection. AB - Chronic HBsAg carriers are known to have a higher risk of hepatitis-related mortality and morbidity when undergoing kidney transplantation. Immunosuppressants might flare up the infection that could be fulminating. Lamivudine and mycophenolate mofetil (MMF) have been shown to be effective in inhibiting replication of hepatitis B virus (HBV). With these two drugs, hepatitis related adverse outcome might be preventable when these patients are being transplanted. Four Chinese adolescents with chronic HBV infection were transplanted in our Department from 1999 to 2001. Immunosuppresants included prednisolone, cyclosporin A and MMF; azathioprine was not used for its potentially liver toxic effect. Prophylactic lamivudine 3 mg/kg and maximum 100 mg daily was given just before transplantation and was continued afterwards. HBV status and liver enzymes were monitored serially. Patients were followed up for 26.0 +/- 10.3 (11-34) months post-transplant and no mortality was reported. All grafts were functioning and no rejection was noted. MMF and lamivudine were well tolerated. Alanine transaminase was only transiently elevated in the first 2 months post-transplant in all patients and became normal afterwards. The patients were clinically well and liver function was normal at the last follow-up. However, HBV DNA became positive in three patients after the transplantation. YMDD mutant HBV was negative in one patient and undeterminable in the other three due to low virus load. In summary, with prophylactic lamivudine and MMF, short term follow-up showed that renal transplant might be feasible and safe in chronic HBV carriers. PMID- 14738299 TI - Immediate tracheal extubation of pediatric liver transplant recipients in the operating room. AB - Keeping patients on mechanical ventilation after orthotopic liver transplantation (OLT) has been a standard anesthetic approach since the first utilization of liver transplantation. Advances in anesthetic management, surgical techniques and patient preparation, in addition to improved postoperative care and the reported advantages of early postoperative tracheal extubation of liver recipients. encouraged us to extubate most recipients at the end of the operation. The aim of the present study is to evaluate the pediatric liver recipients who were extubated immediately at the end of transplantation, in terms of respiratory complications and allograft function during their stay in the ICU. We retrospectively reviewed the records of 40 pediatric recipients who had undergone OLT at the Ege University Organ Transplantation Center between December 1997 and July 2002. Twelve out of 40 patients who had consecutively undergone OLT were extubated immediately at the end of the operation and were included this study. Mean Child Pugh scores of the patients were 9 +/- 2.3 (range 6-12) and the mean PELD score was 23.1 +/- 12.3 (range 7-41). The mean age of the patients was 8.4 +/- 5.2 (range 0.8-16.8 yr). Five of the 12 extubated patients received a cadaveric and seven a living donor liver graft. The mean ICU stay of the patients was 49.1 +/- 24.2 h (6-120 h). No patients required reintubation or mechanical ventilation in the ICU. Respiratory complications diagnosed in the 12 extubated patients were hypercapnia without hypoxemia in three, atelectasis in one and pleural effusion in two. No primary non-function or delayed graft function was detected. The aspartate transaminase (AST), alanine transaminase (ALT) and protrombin time (PT) were normalized within a week. We believe that immediate tracheal extubation in the operating room is a safe procedure for selected cadaveric and living-related liver transplant recipients and will facilitate the patients' recovery and mobilization leading to reduction in complications and a reduced ICU stay. PMID- 14738300 TI - Cognitive and emotional outcome after pediatric liver transplantation. AB - The aim of the study was to evaluate the cognitive and emotional development after pediatric liver transplantation. A total of 21 patients, aged 4-16.9 yr (median 9.6 yr) were tested 1-9 yr (median 4.2 yr) after the transplantation. The pretransplant diagnoses included biliary atresia (eight patients), various metabolic diseases (n = 6), acute liver failure (n = 3), and miscellaneous (n = 4). The cognitive functions were tested with Wechsler preschool and primary scale of intelligence (WPPSI)-R or Wechsler intelligence scale for children (WISC)-III according to age. The Piers-Harris self-concept scale and the evaluation of human figure drawings according to Koppitz were used to detect emotional problems. All tests in all patients were performed by the same psychologist. A significantly lower result on cognitive tests was seen when compared with the expected normal values (p < 0.01). The number of patients with results within or under the lower normal range was higher than expected. Although the mean value of the Piers Harris self-concept scale was normal, there was a large spread within the group. Indicators of emotional problems were found in the human figure drawings of 50% of the patients. To some extent, low cognitive scores coincided with low scores on self-concept scale and indicators of emotional difficulties. We conclude that the high degree of cognitive and emotional problems after liver transplantation is an important argument for routine psychologic follow-up and support in these patients. PMID- 14738301 TI - Psychological functioning of pediatric lung transplant candidates/recipients: a review of the literature. AB - Although lung transplants are performed in children, experience with the pediatric population remains limited. There is growing interest in studying the psychological functioning and quality of life in these patients following transplant. There is a body of literature about quality of life in adult lung transplant recipients, but little is known about how pediatric patients and their families function psychologically after transplant. The current article summarizes the pediatric literature with respect to psychological outcomes for transplant recipients and their parents and points to areas where additional research is needed. PMID- 14738302 TI - Nephrotic syndrome after conversion to alternate day steroids in two children with a history of recurrent FSGS. AB - FSGS is a common indication for kidney transplantation in children. However, transplantation is often complicated by recurrence of FSGS in the transplanted kidney, resulting in nephrotic syndrome and an increased risk of graft loss. Acute treatment strategies for recurrent FSGS include plasmapheresis and increased immunosuppressive therapy. There is little information on the long-term management of immunosuppression in these patients. We describe two children who were successfully treated with plasmapheresis for recurrent FSGS that occurred immediately post-transplant. Nephrotic syndrome reappeared years later when the patients were converted from daily to alternate day prednisone. In children with a history of FSGS, caution is necessary when altering the dosing schedule of prednisone. PMID- 14738303 TI - Fatal pulmonary thromboembolism after a second course of high-dose chemotherapy with autologous peripheral blood stem cell transplantation. AB - We describe a 4-yr-old boy with an occult primitive neuroectodermal tumor, who suffered fatal PTE after a second course of HDC with autologous PBSCT. On day + 52 after a second PBSCT, he was admitted because of respiratory distress. Respiratory failure rapidly progressed and he died within 4 days. The diagnosis of PTE was confirmed by a lung perfusion study with technetium-99m macroaggregated albumin, but too late to allow treatment. Although rare, PTE must be recognized as an important differential diagnosis when respiratory symptoms are observed after HDC with PBSCT. PMID- 14738304 TI - Basiliximab monotherapy following B-cell lymphoma after pediatric liver transplantation and anti-CD20 therapy. AB - The chimeric, monoclonal antibody basiliximab inhibits the proliferation and clonal expansion of activated T cells. To date basiliximab has been used only in combination with other immunosuppressive agents for rejection prophylaxis after solid organ transplantation. An infant underwent liver transplantion (LTx) at the age of 5 months because of biliary atresia. The primary immunosuppression consisted of cyclosporine and prednisolone. As a result of a steroid resistant rejection episode on day 26 post-LTx we had to switch the initial immunosuppressive regiment to tacrolimus and steroids. Because of severe cholestasis and assumed impaired enteral resorption we were forced to administer an unusually high dosage (2 mg/kg/day) of tacrolimus. Four months after LTx an intestinal B-cell lymphoma was diagnosed when the patient suffered from a small bowel perforation. After stopping the immunosuppressive medication we started treatment with the anti-CD20 monoclonal antibody rituximab for B-cell depletion. During the 12 wk no B cells were detectable in the peripheral blood by flow cytometry. In this setting we started a monotherapy with repetitive doses of basiliximab for immunosuppression. During the following course there was no further rejection and no recurrence of the tumor. From this experience we conclude that monotherapy with basiliximab after LTx and anti-CD20 treatment for B-cell lymphoma is efficient and safe. PMID- 14738305 TI - End-stage renal disease after high-dose carboplatinum in preparation of autologous stem cell transplantation. AB - Stem cell transplantation is one therapy employed in the management of children with high-risk solid tumors. However, this therapy is not without risk, having been associated with multiple end-organ toxicities. Both acute renal failure and chronic renal insufficiency have been reported in marrow transplant recipients, primarily in the context of the use of calcineurin inhibitors and radiation therapy. This report reviews our experience in managing an adolescent with metastatic Ewing's sarcoma who developed rapid progression to end-stage renal disease following a pretransplant conditioning regimen with high-dose carboplatinum. She had not received radiation or prior cisplatinum therapy. The possible reasons for the patient's highly unusual course and recommendations on ways to prevent this complication are discussed. PMID- 14738306 TI - Bordetella bronchiseptica infection in pediatric lung transplant recipients. AB - Bordetella bronchiseptica are small, pleomorphic Gram-negative coccobacilli which are commensal organisms in the upper respiratory tract of many wild and domestic animals ('kennel cough' in dogs). While it is common for health care providers to ask about exposure to ill family/friends, most do not routinely inquire about the health or immunization status of household pets. We report two cases of B. bronchiseptica pneumonia in lung transplant recipients [cystic fibrosis (CF); ages 10 and 15 yr; one male] who contracted B. bronchiseptica from pet dogs. We compared their course and outcome to four children (two CF, one congenital heart disease and one Duchenne's muscular dystrophy; four males, age range 6 months to 14 yr) with B. bronchiseptica cultured from the respiratory tract. Two of the four patients also acquired their illnesses from pet dogs and two from unknown sources. One lung transplant recipient expired from progressive respiratory failure. We conclude that B. bronchiseptica can cause serious infections in both immunosuppressed and immunocompetent children. We speculate that a detailed history of exposure to ill pets (particularly dogs), and the immunization status of all pets should be included in the routine evaluation of all pediatric transplant recipients. PMID- 14738307 TI - The carboxylesterase gene family from Arabidopsis thaliana. AB - Carboxylesterases hydrolyze esters of short-chain fatty acids and have roles in animals ranging from signal transduction to xenobiotic detoxification. In plants, however, little is known of their roles. We have systematically mined the genome from the model plant Arabidopsis thaliana for carboxylesterase genes and studied their distribution in the genome and expression profile across a range of tissues. Twenty carboxylesterase genes (AtCXE) were identified. The AtCXE family shares conserved sequence motifs and secondary structure characteristics with carboxylesterases and other members of the larger alpha/beta hydrolase fold superfamily of enzymes. Phylogenetic analysis of the AtCXE genes together with other plant carboxylesterases distinguishes seven distinct clades, with an Arabidopsis thaliana gene represented in six of the seven clades. The AtCXE genes are widely distributed across the genome (present in four of five chromosomes), with the exception of three clusters of tandemly duplicated genes. Of the interchromosomal duplication events, two have been mediated through newly identified partial chromosomal duplication events that also include other genes surrounding the AtCXE loci. Eighteen of the 20 AtCXE genes are expressed over a broad range of tissues, while the remaining 2 (unrelated) genes are expressed only in the flowers and siliques. Finally, hypotheses for the functional roles of the AtCXE family members are presented based on the phylogenetic relationships with other plant carboxylesterases of known function, their expression profile, and knowledge of likely esterase substrates found in plants. PMID- 14738308 TI - Phylogenetic analysis of vertebrate fibrillar collagen locates the position of zebrafish alpha3(I) and suggests an evolutionary link between collagen alpha chains and hox clusters. AB - Type I collagen in tetrapods is usually a heterotrimeric molecule composed of two alpha1 and one alpha2 chains. In some teleosts, a third alpha chain has been identified by chromatography, suggesting that type I collagen should also exist as an alpha1(I)alpha2(I)alpha3(I) heterotrimer. We prepared, from zebrafish, three distinct cDNAs identified to be those of the collagen alpha1(I), alpha2(I), and alpha3(I) chains. In this study on the evolution of fibrillar collagen alpha chains and their relationships, an exhaustive phylogenetic analysis, using vertebrate fibrillar collagen sequences, showed that each alpha chain constitutes a monophyletic cluster. Results obtained with the newly isolated sequences of the zebrafish showed that the alpha3(I) chain is phylogenetically close to the alpha1(I) chain and support the hypothesis that the alpha3(I) chain arose from a duplication of the alpha1(I) gene. The duplication might occur during the duplication of the actinopterygian genome, soon after the divergence of actinopterygians and sarcopterygians, a hypothesis supported by the demonstration of a syntenic evolution between a set of fibrillar collagen genes and Hox clusters in mammals. An evolutionary scenario is proposed in which phylogenetic relationships of the alpha chains of fibrillar collagens of vertebrates could be related to Hox cluster history. PMID- 14738309 TI - EVG, the remnants of a primordial bilaterian's synteny of functionally unrelated genes. AB - Extant genomes are the result of repeated duplications and subsequent divergence of primordial genes that assembled the genomes of the first living beings. Increased information on genome maps of different species is revealing conserved syntenies among different vertebrate taxa, which allow to trace back the history of current chromosomes. However, inferring neighboring relationships between genes of more primitive genomes has proven to be very difficult. Most often, the ancestral arrangements of genes have been lost by multiple histories of internal duplications, chromosomal breaks, and large-scale genomic rearrangements. Here we describe a gene arrangement of nonrelated genes that seems to have endured evolution, at least from the separation of the two major clades of bilateria: deuterostomia and protostomia, approximately 1 billion years ago. In its simplest conception, this gene cluster, named EVG, groups the genes for a glucose transporter, an enolase, and a vesicle-associated membrane protein (VAMP). EVG might represent the evolutionary remnants of the gene organization of an ancient bilaterian genome. PMID- 14738310 TI - Concerted changes in the nucleotide sequences of the intragenic promoter regions of eukaryotic genes for tRNAs of all specificities. AB - RNA polymerase III promoter is located within the coding region in all eukaryotic tRNA genes, whereas in prokaryotic tRNA genes, the promoter is located upstream of the transcription initiation site. We analyzed the nucleotide sequence context of the A and B boxes of RNA polymerase III promoters from different unicellular eukaryotes, plants, and animals and the homologous sequences in the tRNA genes of prokaryotic species (Archaea, Eubacteria). The long and short sequence variants of the A box are nonrandomly distributed across different types of the eukaryotic tRNA genes. In contrast, the sequences from the prokaryotic counterparts of the long and short variants are randomly scattered among prokaryotic tRNA genes. The sequence diversity of the 3' portion of the A box in the eukaryotic tRNA genes was sharply reduced compared to that of the homologous segments of the prokaryotic tRNA genes. Analysis of the frequencies of oligonucleotide variants from different parts of the A box suggested that the prominent changes in the structure of the A box occurred at about the same time in all tRNA genes during the prokaryote eukaryote evolutionary transition. PMID- 14738311 TI - Codon usage decreases the error minimization within the genetic code. AB - The genetic code is not random but instead is organized in such a way that single nucleotide substitutions are more likely to result in changes between similar amino acids. This fidelity, or error minimization, has been proposed to be an adaptation within the genetic code. Many models have been proposed to measure this adaptation within the genetic code. However, we find that none of these consider codon usage differences between species. Furthermore, use of different indices of amino acid physicochemical characteristics leads to different estimations of this adaptation within the code. In this study, we try to establish a more accurate model to address this problem. In our model, a weighting scheme is established for mistranslation biases of the three different codon positions, transition/transversion biases, and codon usage. Different indices of amino acids' physicochemical characteristics are also considered. In contrast to pervious work, our results show that the natural genetic code is not fully optimized for error minimization. The genetic code, therefore, is not the most optimized one for error minimization, but one that balances between flexibility and fidelity for different species. PMID- 14738312 TI - Nonrandom intragenic variations in patterns of codon bias implicate a sequential interplay between transitional genetic drift and functional amino acid selection. AB - Although most codon third bases appear to be functionless, the synonymous codons so defined exhibit a strikingly nonrandom distribution (codon bias) within human and other genes. To examine this phenomenon further, we generated a database of DNA sequences encoding human transmembrane cell-surface receptor proteins. Using this database we show here that the guanine and cytosine content of codon third bases (GC3) varies intragenically with the nature of the specified receptor domains (transmembrane > extracellular > intracellular domains; p < 0.001), the phenotype of the encoded amino acids (hydrophobic > hydrophilic > neutral amino acids; p < 0.001), and the receptor affiliation of the transmembrane (G-protein coupled receptors > receptor tyrosine kinases; p < 0.001). Within gene regions specifying transmembrane domains, GC3 declines as domain functionality becomes redundant with increasing hydrophobicity (p < 0.001). Codons containing the second-base cytosine (XCZ, which encodes neutral amino acids) are selectively depleted of third-base adenine content (A3: XCA codons) when encoding transmembrane domain residues, consistent with positive selection for transitional mutation of XCG to XTG (which encodes hydrophobic amino acids) rather than to the synonymous XCA. Supporting this XCG --> XTG mechanism of codon bias, the G3:A3 ratio of codons specifying the transmembrane amino acid glycine (GGZ) is intermediate between that of its functional homolog alanine (GCZ) and that of hydrophobic valine (GTZ), even though the C3:T3 ratios are similar. Conversely, nearest-neighbor analysis of third bases 5' to codons specifying valine and leucine (CTZ) confirms a significant difference in C3:T3 but not G3:A3 ratios (i.e., C3/G1 --> T3/G1 > C3/A1; p < 0.001), consistent with the functionally advantageous retention of hydrophobic residues. These data raise the possibility that patterns of intragenic codon bias reflect a balance between negative and positive selection, suggesting in turn that analysis of codon third base usage may help to predict the functional significance of encoded products. PMID- 14738313 TI - Interisland mutation of a novel phospholipase A2 from Trimeresurus flavoviridis venom and evolution of Crotalinae group II phospholipases A2. AB - Trimeresurus flavoviridis (Crotalinae) snakes inhabit the southwestern islands of Japan: Amami-Oshima, Tokunoshima, and Okinawa. Affinity and conventional chromatographies of Amami-Oshima T. flavoviridis venom led to isolation of a novel phospholipase A2 (PLA2). This protein was highly homologous (91%) in sequence to trimucrotoxin, a neurotoxic PLA2, which had been isolated from T. mucrosquamatus (Taiwan) venom, and exhibited weak neurotoxicity. This protein was named PLA-N. Its LD50 for mice was 1.34 microg/g, which is comparable to that of trimucrotoxin. The cDNA encoding PLA-N was isolated from both the Amami-Oshima and the Tokunoshima T. flavoviridis venom-gland cDNA libraries. Screening of the Okinawa T. flavoviridis venom-gland cDNA library with PLA-N cDNA led to isolation of the cDNA encoding one amino acid-substituted PLA-N homologue, named PLA-N(O), suggesting that interisland mutation occurred and that Okinawa island was separated from a former island prior to dissociation of Amami-Oshima and Tokunoshima islands. Construction of a phylogenetic tree of Crotalinae venom group II PLA2's based on the amino acid sequences revealed that neurotoxic PLA2's including PLA-N and PLA-N(O) form an independent cluster which is distant from other PLA2 groups such as PLA2 type, basic [Asp49]PLA2 type, and [Lys49]PLA2 type. Comparison of the nucleotide sequence of PLA-N cDNA with those of the cDNAs encoding other T. flavoviridis venom PLA2's showed that they have evolved in an accelerated manner. However, when comparison was made within the cDNAs encoding Crotalinae venom neurotoxic PLA2's, their evolutionary rates appear to be reduced to a level between accelerated evolution and neutral evolution. It is likely that ancestral genes of neurotoxic PLA2's evolved in an accelerated manner until they had acquired neurotoxic function and since then they have evolved with less frequent mutation, possibly for functional conservation. PMID- 14738314 TI - A novel family of unconventional actins in volvocalean algae. AB - The unicellular green alga Chlamydomonas reinhardtii has two actin genes, one encoding a conventional actin (90% amino acid identity with mammalian actin), the other a highly divergent actin (64% identity) named novel actin-like protein (NAP). To see whether the presence of conventional and unconventional actins in a single organism is unique to C. reinhardtii, we searched for genomic sequences related to the NAP sequence in several other species of volvocalean algae. Here we show that Chlamydomonas moewusii and Volvox carteri also have, in addition to a conventional actin, an unconventional actin similar to the C. reinhardtii NAP. Analyses of the deduced protein sequences indicated that the NAP homologues form a distinct group derived from conventional actin. PMID- 14738315 TI - Molecular phylogeny of the Drosophila melanogaster species subgroup. AB - Although molecular and phenotypic evolution have been studied extensively in Drosophila melanogaster and its close relatives, phylogenetic relationships within the D. melanogaster species subgroup remain unresolved. In particular, recent molecular studies have not converged on the branching orders of the D. yakuba-D. teissieri and D. erecta-D. orena species pairs relative to the D. melanogaster-D. simulans-D. mauritiana-D. sechellia species complex. Here, we reconstruct the phylogeny of the melanogaster species subgroup using DNA sequence data from four nuclear genes. We have employed "vectorette PCR" to obtain sequence data for orthologous regions of the Alcohol dehydrogenase (Adh), Alcohol dehydrogenase related (Adhr), Glucose dehydrogenase (Gld), and rosy (ry) genes (totaling 7164 bp) from six melanogaster subgroup species (D. melanogaster, D. simulans, D. teissieri, D. yakuba, D. erecta, and D. orena) and three species from subgroups outside the melanogaster species subgroup [D. eugracilis (eugracilis subgroup), D. mimetica (suzukii subgroup), and D. lutescens (takahashii subgroup)]. Relationships within the D. simulans complex are not addressed. Phylogenetic analyses employing maximum parsimony, neighbor-joining, and maximum likelihood methods strongly support a D. yakuba-D. teissieri and D. erecta-D. orena clade within the melanogaster species subgroup. D. eugracilis is grouped closer to the melanogaster subgroup than a D. mimetica-D. lutescens clade. This tree topology is supported by reconstructions employing simple (single parameter) and more complex (nonreversible) substitution models. PMID- 14738317 TI - Molecular evolution of cycloidea-like genes in Fabaceae. AB - The cycloidea (CYC) gene controls floral symmetry in snapdragon (Antirrhinum majus). We investigated the evolution of CYC-like genes in some species of legumes that have zygomorphic flowers. Two to four CYC-like genes were isolated from a single species. The results of NJ and ML analyses indicate that CYC-like genes in legumes group into two monophyletic clades; one group consists of eight CYC-like genes (Clade 1) and the other contains three CYC-like genes and TB1 of maize (Clade 2). These phylogenetic trees and the Shimodaira-Hasegawa test suggest that Clade 1 is a sister of the original CYC group (Clade 3). Moreover, the result of the GeneTree analysis showed that the CYC-like genes experienced repeated duplication events during the evolution of legumes. We herein speculate as to the role of CYC-like genes in legumes and discuss the evolutionary processes that these genes have undergone. PMID- 14738316 TI - Mitochondrial genome of Ciona savignyi (Urochordata, Ascidiacea, Enterogona): comparison of gene arrangement and tRNA genes with Halocynthia roretzi mitochondrial genome. AB - The complete nucleotide sequence of the urochordate Ciona savignyi (Ascidiacea, Enterogona) mitochondrial (mt) genome (14,737 bp) was determined. The Ciona mt genome does not encode a gene for ATP synthetase subunit 8 but encodes an additional tRNAGly gene (anticodon UCU), as is the case in another urochordate, Halocynthia roretzi (Ascidiacea, Pleurogona), mt genome. In addition, the Ciona mt genome encodes two tRNAMet genes; anticodon CAT and anticodon TAT. The tRNACys gene is thought to lack base pairs at the D-stem. Thus, the Ciona mt genome encodes 12 protein, 2 rRNA, and 24 tRNA genes. The gene arrangement of the Ciona mt genome differs greatly from those of any other metazoan mt genomes reported to date. Only three gene boundaries are shared between the Halocynthia and the Ciona mt genomes. Molecular phylogenetic analyses based on amino acid sequences of mt protein genes failed to demonstrate the monophyly of the chordates. PMID- 14738318 TI - The evolution of parasite recognition genes in the innate immune system: purifying selection on Drosophila melanogaster peptidoglycan recognition proteins. AB - Genes involved in the recognition of parasites by the acquired immune system are often subject to intense selection pressures. In some cases, selection to recognize a diverse range of parasites has resulted in high levels of polymorphism, while elsewhere the protein sequence has changed rapidly under directional selection. We tested whether parasite recognition genes in the innate immune system show similar patterns of evolution. We sequenced seven peptidoglycan recognition protein genes (PGRPs) from 12 lines of Drosophila melanogaster and one line of D. simulans and used a variety of tests to determine whether the observed mutations were selectively neutral. We were unable to detect either balancing or directional selection. This suggests that the molecular cues used by insects to detect parasites are highly conserved and probably under strong functional constraints which prevent their evolving to evade the host immune response. Therefore, interactions between these genes are unlikely to be the focus of host-parasite coevolution, at least in Drosophila. We also found evidence of gene conversion occurring between two genes, PGRP-SC1A and PGRP-SC1B. PMID- 14738319 TI - Gene-based deafness research: ion transport and hearing. AB - The cochlea is a sensory organ that converts physical (sound) stimulation into electrical signals. This process is fundamentally and substantially based upon the ion transport system. Here, I summarize the physiological and molecular biological aspects of transporters, channels and receptors expressed in the cochlea. With reference to these findings, recent advances in genetic research on hereditary deafness are discussed. PMID- 14738320 TI - Altered distribution of metaplastic Paneth, gastrin and pancreatic acinar cells in atrophic gastritic mucosa with endocrine cell lesions. AB - The mechanism of progression from gastric endocrine cell hyperplasias (ECHs) to carcinoid tumor (GCT) is still unknown. In these lesions, the distribution of metaplastic Paneth, gastrin and pancreatic acinar cells developing due to consequences of corporal mucosal atrophy has not been investigated in detail. In this study, 33 gastric endoscopic biopsies with endocrine cell lesions were examined. In all cases except 6 with solitary GCT, complete-type (small intestine) intestinal metaplasia (IM) with Paneth cells was observed. The density of lysozyme-positive Paneth cells in IMs in cases with GCTs was less than those in ECH alone. The density of gastrin-positive cells in IMs and average number of micronodules of ECHs were similar. Pancreatic acinar metaplasia (PAM) was observed in 6 cases of GCTs with ECH. The size of GCTs with ECH was smaller than those without ECH. By image analysis, the percentage of Ki67 (MIB-1, proliferation marker) expressing cells of GCTs with ECH was 5.1+/-0.6%, and GCT without ECH 7.8+/-1%. Our results indicate that few Paneth cells and many PAMs in atrophic corporal mucosa are seen more frequently in cases of GCTs with ECH, compared to those in ECH alone. Gastrin-positive cells in the corporal IM may stimulate enterochromaffin-like (ECL) cells, which may induce hyperplasia, dysplasia or neoplasia by augmenting the effects of hypergastrinemia through a paracrine mechanism on local gastrin-sensitive cells. PMID- 14738321 TI - Susceptibility patterns of enterococci causing infections. AB - Enterococci are among the common organisms associated with hospital-acquired infections. We examined in vitro activities of different antibiotics to 103 enterococcal isolates. Minimal inhibitory concentrations (MICs) of penicillin G, ampicillin, gentamicin, ciprofloxacin, ofloxacin, levofloxacin, grepafloxacin, trovafloxacin and gemifloxacin were determined by broth microdilution testing method. Among the isolates 71 (69%) were identified as E. faecalis and 32 (31%) as E. faecium. While over 75% of E. faecium isolates were resistant to penicillin and ampicillin, approximately 25% of E. faecalis isolates were resistant to penicillin and ampicillin. None of the E. faecalis and E. faecium isolates were resistant to vancomycin. While 17 (52%) of E. faecium isolates exhibited high level gentamicin resistance (HLGR), high level streptomycin resistance (HLSR) was detected in 24 (74%) of the isolates. In contrast, HLGR and HLSR rates for E. faecalis were 14 (20%) and 22 (31%), respectively. Both HLGR and HLSR were detected with higher frequency in ampicillin resistant isolates. Among fluoroquinolones, gemifloxacin and trovafloxacin were the most potent antibiotics tested. There was no increase in MIC90 values of the fluoroquinolones in ampicillin resistant isolates in comparison with ampicillin susceptible isolates. Our data suggest newer fluoroquinolones would be good alternative agents to use especially for combination drug therapy where enterococci with ampicillin resistance and HLAR are prevalent. PMID- 14738322 TI - Effects of alcohol consumption on hepatocellular injury in Japanese men. AB - To clarify the effects of alcohol consumption on hepatocellular injury, we examined aspartate and alanine aminotransferases (AST and ALT), and gamma glutamyltransferase (GGT), together with weekly alcohol consumption calculated from a self-rating questionnaire, in 1113 Japanese salesmen. The thresholds of associations between alcohol consumption and liver markers were estimated by the benchmark dose (BMD) method. The AST, ALT and GGT were positively correlated with alcohol intake (p<0.001), as well as age and body mass index (BMI); the relations to alcohol were statistically significant even when controlling for age, BMI and smoking habit. Although the AST and GGT were associated with four types of alcoholic beverage (p<0.01), it was only whiskey that had close relation to the ALT (p<0.05). The thresholds of alcohol consumption (ethanol g/week), i.e., 95% lower confidence limits of the BMD, were 362 for AST, 660 for ALT, and 252 for GGT. The thresholds for GGT and AST in Japanese men seem to be somewhat higher than those reported in Western countries. It is suggested that hepatocellular injury (i.e., AST elevation) in Japanese men may emerge at the ethanol level of more than 50 g/day. PMID- 14738323 TI - STAT1 knockout mice are highly susceptible to pulmonary mycobacterial infection. AB - This study was designed to determine the roles of STAT1 protein in defense against mycobacterial infection. Airborne infection of STAT1 knockout (KO) mice with a Mycobacterium tuberculosis Kurono strain induced multiple necrotic lesions in lungs, spleen and liver, while that in wild-type (WT) mice did not. The STAT1 KO mice succumbed to mycobacterial infection by the 35th day after infection. Compared with the levels in WT mice, inducible nitric oxide synthase (iNOS), tumor necrosis factor-alpha, interferon-gamma and IL-12 mRNA levels were significantly lower in the lung of STAT1 KO mice. Interestingly, granulomatous lesion development in STAT1 KO mice was inhibited significantly by treatment with exogenous recombinant murine IL-12. Therefore, STAT1 regulates IL-12 expression and appears to be a critical transcription factor in controling mycobacterial infection. PMID- 14738324 TI - Effects of open vs. laparoscopic cholecystectomy on oxidative stress. AB - Elective laparoscopic cholecystectomy is established as the treatment of choice for symptomatic cholecystolithiasis and is now proposed for the treatment of acute cholecystitis. The aim of this study is to evaluate biochemical aspects of open (OC) and laparoscopic cholecystectomy (LC). We measured the levels of malondialdehyde (MDA) and the levels of nitrite+nitrate as stable end products of nitric oxide (NO). MDA and nitrite+nitrate levels were increased at both surgical procedures compared to preoperative period, but the rise was more significant in OC than LC. These results showed that both OC and LC caused an increase in oxidative stress. However LC caused significantly less oxidative stress and the changes during surgery returned to preoperative values after LC in a shorter period. The beneficial effects of laparoscopic surgery may be related, partially, to less oxidative stress in the immediate postoperative period. PMID- 14738325 TI - Effect of intra-arterial cisplatin on multiple liver metastases from rectal cancer associated with ulcerative colitis. AB - We report a patient with synchronous multiple liver metastases from rectal cancer associated with ulcerative colitis. Because the liver tumors were unresectable, we performed total proctocolectomy and hepatic intra-arterial cisplatin infusion with systemic oral administration of fluorouracil. A complete response was obtained. The patient is alive without sign of recurrence 5 years postoperatively. Hepatic intra-arterial administration of cisplatin should be considered in the treatment of unresectable liver metastases from colorectal cancer. PMID- 14738326 TI - Transmission electron microscopy of Schistosoma mansoni cercariae treated with hinokitiol (beta-thujaplicin), a compound for potential skin application against cercarial penetration. AB - Since skin is the only route of entry of the parasite in schistosomiasis patients, intervention at the level of skin penetration should control the infection. Several compounds were screened for their ability to protect against cercarial penetration. Hinokitiol (beta-thujaplicin) was found to have a significant cercaricidal effect in vitro, although there is no information on its cercaricidal mechanisms. To study the kinetics of morphological changes in Schistosoma mansoni associated with exposure to hinokitiol in vitro, cercariae were incubated in media containing hinokitiol at different concentrations and examined by transmission electron microscopy (TEM). TEM revealed that ultrastructural changes occurred by 15 minutes post exposure, at a concentration of 25 microg/ml. Degenerative changes involving both tegument and deeper parenchymal structures were progressive with duration of exposure at the concentration of 50 microg/ml. These structural changes may account for the inability of hinokitiol-treated cercariae to infect the host. PMID- 14738327 TI - Identification of sounds from traffic. AB - Listeners' ability to identify road-traffic, aircraft, or train sounds in environmental sound recordings was studied in a psychoacoustical experiment involving 16 participants. In free-labeling identification, excerpt traffic sounds were described in terms of "object" (sound-producing source) rather than in terms of perceptual attribute. The main sounds identified were traffic sounds, but a few references were also made to machine-related or water-related sources. Sounds from aircraft were easier to identify than the sounds from trains, which in turn were easier to identify than the sounds from road-traffic. This identification order was confirmed in multiple-choice and dominant-source identification tasks. Compared to free-labeling, multiple-choice identifications produced considerably more false alarms, i.e., identification of a sound source not present. For multiple-choice, several sound sources were particularly identified in the excerpt of road-traffic and train sounds although the (recorded) sound was typically clearly discerned in the joint dominant-source identification task. A comparison of the acoustic properties of the traffic sounds suggested that spectral rather than temporal cues were used in sound source identification. PMID- 14738328 TI - Perceptions about exercise and intrinsic motivation of students attending a health-related physical education program. AB - The present study examined effects of sex, attitude towards physical activity, perceived barriers for participation in physical activity, and students' perception of their parents' participation in physical activity on the intrinsic motivation of students participating in a health related program in physical education. 643 students (303 boys and 340 girls) responded to questionnaires measuring intrinsic motivation, attitudes towards physical activity, perceived barriers to exercise and perceived parents' participation in physical activity. Mean age was 12.9 yr. (SD=1.2, range 11-14 years). Analysis indicated that students' intrinsic motivation towards the program was influenced by perceived barriers to exercise. Sex, attitudes towards physical activity, and perceived parents' participation in physical activity seem to be less important. PMID- 14738329 TI - Expression of emotions in dance: relation between arm movement characteristics and emotion. AB - This study was designed to investigate the relations between emotional expression and the movement characteristics. For this purpose, we used kinematic data related to three factors of the movement characteristics: Speed, Force, and Directness. In Exp. 1, we examined how the dancers expressed emotions when they used a certain body action and body part, and how they altered the movement characteristics. In Exp. 1, 10 female dancers were instructed to express three emotions, joy, sadness, and anger, by altering arm-movement characteristics. Analysis of variance indicated that the three exhibited emotional expressions had different movement characteristics. Discriminant analysis indicated that kinematic data for evaluation of movement characteristics are useful for discrimination of the three emotional expressions in dance. In Exp. 2, we investigated how naive observers perceived the type of emotion from the arm movement characteristics. Analysis of variance showed that 22 observers accurately perceived each emotion distinguished from other emotions. Multiple regression analysis showed that specific movement characteristics influenced the perception of particular emotion. PMID- 14738330 TI - Using the Delphi technique to select items for use in assessing patients' satisfaction with dysphagia services. AB - Items for potential use in a survey for the assessment of patients' satisfaction with dysphagia services was developed using the Delphi technique. A panel of speech-language pathologists was provided with 105 potential survey items. They were asked to rate the items with respect to perceived importance in assessing patients' satisfaction. 45 items were rated as "very important." Items concerning the issues of information giving and the conduct and characteristics of the clinician were most common among the selected items. No items focusing on accessibility of services or quality of the treatment environment were in the "very important" group. PMID- 14738331 TI - What difference does it make? Implications of the size of the difference between the means of two groups. AB - This study employed a model with two normal distributions of scores to study group differences and similarities as a function of the distance between group means. The mean of one distribution was systematically moved further and further away from the mean of the other. A table was developed of associated measures of group overlap and separation, e.g., r, eta, percent correct classification, percentage of overlap, for several levels of d' (difference between means expressed in standard deviation units). The implications of various d' values for the selection of extreme groups, e.g., top 1% of scorers, were discussed. It was noted that even small differences between means could result in an extremely disproportionate inclusion of members from the two original groups in a high scoring subgroup. PMID- 14738332 TI - Factors affecting bone mineral density in the general adult female population in Japan. AB - The factors affecting bone mineral density were evaluated in the general adult female population of Shibukawa City, Japan. Bone mineral density at the radius site of the wrist was measured by dual-energy X-ray absorptiometry. 749 women between the ages of 39 and 52 years were selected from the general population by a complete enumeration method. Age, Body Mass Index, age of menarche, past history of treated chronic disease, symptoms of lower back pain, exercise habits which included any exercise at all, medication for chronic disease, diet and history of dieting, smoking habits, alcohol consumption, menstruation, childbirth, breast-feeding, content of meals which was taken at least once per week, and past history of bone fracture were used to predict bone mineral density compared with the average values for young females. The mean bone mineral density of subjects with no experience of low back pain was significantly lower than that of subjects reporting lower back pain symptoms both at rest and with moving (p<.05). Body Mass Index was significantly related to bone mineral density (r=.43, p<.001). Aging, past history of bone fracture, low Body Mass Index, no experience of low back pain at rest, menopause, and no history of dieting were significantly positively associated with bone mineral density using a multiple regression analysis. PMID- 14738333 TI - Instructions emphasizing velocity, accuracy, or both in performance and kinematics of overarm throwing by experienced team handball players. AB - On motor tasks like the overarm throw, velocity and accuracy are two important parameters of performance that may be incompatible and require different strategies in the execution of the motor task. The purpose was to investigate the effects of instruction (emphasizing velocity, accuracy, or both) on performance and kinematics of overarm throwing. Results for 9 experienced male team handball players (M age = 24 +/- 2.2 yr.) showed type of instruction affected the maximal ball velocity. The difference in ball velocity reflected the significant difference in maximal linear velocity of the wrist, elbow, and hip segments together with their absolute timing before ball release. The subjects did not seem to change their throwing technique, i.e., the relative timing of movement initiation of the different body segments. PMID- 14738334 TI - Plasma cholesterol levels and Irlen syndrome: preliminary study of 10- to 17-yr. old students. AB - The preliminary study investigated metabolic anomalies in children and teenagers with Irlen Syndrome, particularly in relation to the levels of n-3 and n-6 essential fatty acids, plasma cholesterol levels, and the relative abundance of plasma saturated fatty acids. The experimental group involved 13 subjects with Irlen Syndrome (M=13.3 yr., SD=2.5 yr.), with a comparison group of 16 age- and sex-matched controls (M=13.8 yr., SD=2.4 yr.). The Irlen Syndrome group were selected from people referred for help with reading and writing problems. The control group were primarily recruited from the general public. All subjects were screened for symptoms of the syndrome using the Scotopic Sensitivity Syndrome Screening Manual. Samples of whole blood were collected and plasma extracted. Metabolites were compared using the Student t test. There were no differences in n-3 and n-6 essential fatty acids between Irlen Syndrome and control groups, although the former group had lower mean levels in most of these essential fatty acids. Total plasma cholesterol level was significantly decreased for the Irlen Syndrome group, and there was a significant increase in the relative abundance of the odd-chain fatty acid, heptadecanoic acid. The differences in heptadecanoic acid may have implications for altered membrane function and neurotransmission. The differences in plasma cholesterol levels, as well as heptadecanoic acid, may also point to the presence of viral or bacterial infection. PMID- 14738335 TI - Web-based communication patterns of physical education teachers. AB - Web-based bulletin board use and topics were examined for 48 physical education student teachers during their 16-wk. student teaching experience. Analysis showed web-based communication was positive in fostering discussion from peers, feedback, and support. PMID- 14738336 TI - Scores on test of gross motor development of children with dysphasia: a pilot study. AB - The purpose of this preliminary investigation was to assess the gross motor skill and trainability of children with dysphasia using the Test of Gross Motor Development. 27 children from 7 to 12 years (M=9.6 yr., SD=1.3) from four different classes in one special school participated. The gross motor skill scores of a sample of children with dysphasia, their trainability of the gross motor skills, and the intrarater reliability of the test were examined. Based on the videotaped performances the intrarater reliability was .93 to .97 for total scores, but lower for individual test items. Mean motor skill scores of these children with dysphasia were interpreted as 'poor' relative to those of the standardization and the reference group. The 8-wk. intervention showed statistically significant improvement from pre- to posttest values. Mean locomotor skills improved from 2.7 (SD=.4) to 2.8 (SD=.4), and the object control skills from 2.4 (SD=.7) to 2.8 (SD=.7). It appears these children with language disorders do not possess a similar basic gross motor level as their peers without language problems. PMID- 14738337 TI - Validation with a French sample of the four scales of Switras's Survey of Mental Imagery. AB - The present study investigated the psychometric properties of the Switras questionnaire, the Survey of Mental Imagery, with a French sample of 162. Only Form A and four subtests of the questionnaire were given. The present factor analyses confirm the factorial composition observed by Switras in 1978. Three subcapacities of imagery were evaluated by these subtests: controllability, vividness, and formation in four specific modalities of imagery, namely, visual, auditory, somesthetic, and kinaesthetic. PMID- 14738338 TI - Survey of preferences for feedback style in high school athletes. AB - This study asked high school athletes what kinds of desirable traits or styles they would like to see in coaches when they provided correct feedback. High school athletes (M age=14.5 yr., SD=2.1), 43 girls and 70 boys, in both individual and team sports were recruited from 2 junior high and 2 senior high schools. A questionnaire measuring preferences for feedback style was developed with 12 9-point Likert scale items. Each item dealt with one aspect of feedback style. The survey covered several characteristics of feedback style: character, timing, frequency, precision, intent, informational style versus controlling style, descriptive versus prescriptive, and context. Analyses of variance, performed on responses to each question to assess the effects of sex, age, and sport, indicated high uniformity in responses to the qualities of feedback style, with no significant differences for sex, age, or sport. PMID- 14738339 TI - Changes in self-reported incidence of nocturnal bruxism in college students: 1966 2002. AB - The frequencies of self-reported nocturnal bruxism in four large samples of college students were compared to assess change in the incidence of this disorder since 1966. Analysis showed incidence of nocturnal bruxism increased from 5.1% to 22.5% over this period. However, the increase from our 1989 sample was not statistically significant. PMID- 14738340 TI - Left- versus right-hand tracking performance by right-handed boys and girls: examination of performance asymmetry. AB - This study compared left- versus right-hand performance within healthy, right handed, 8- or 9-yr.-old boys and girls on a finger-movement tracking task. 38 boys and 38 girls were randomly assigned to use either the left hand first and right hand second or vice versa in tracking a sine wave target with extension and flexion movements of the index finger. The data were analyzed with a three-way analysis of variance with repeated measures followed by pair-wise comparisons with a Bonferroni correction. Analysis yielded a significant hand x test interaction and a significant improvement for subjects tracking with the right hand on Test 1 and left hand on Test 2. No significant change occurred for subjects tracking with the left hand on Test 1 and right hand on Test 2. No interaction was observed with sex as a factor. This study suggests that asymmetry of performance favoring the left hand occurs in right-handed boys and girls during finger-movement tracking. PMID- 14738341 TI - Relationship between psychological type and sensitivity to anxiety. AB - The Myers-Briggs Type Indicator and the Anxiety Sensitivity Index were administered to 143 undergraduate students. Although there was no significant difference between Extraverts and Introverts in Anxiety Sensitivity scores, those classified as Sensing and Thinking types exhibited higher mean scores on Anxiety Sensitivity than those classified as Intuitive and Feeling types. These findings suggest that knowledge of psychological type may be helpful in predicting sensitivity to anxiety. PMID- 14738342 TI - Home field advantage: new stadium construction and team performance in professional sports. AB - To identify the relations between new stadiums and home team performance and attendance for professional baseball (MLB) (n=14), basketball (NBA) (n=13), and football (NFL) (n=25) teams in the USA since 1950 dependent t tests assessed significance of increases in attendance in both MLB and the NBA and a significantly improved home winning percentage in MLB following the building of new stadiums. Implications include a better understanding of the rationales used by owners, fans, and players for building new stadiums. PMID- 14738343 TI - Sign language and Chinese characters on visual-spatial memory: a literature review. AB - Long-term use of a sign language may enhance visual abilities. Deaf signers consistently outscore their hearing nonsigning counterparts on visual memory tests. In a similar vein, the long-term use of a logographic script, Chinese characters appears to boost visual memory. Both learning a sign language and mastering Chinese characters involve a relatively complex task of visual recognition and memory. This review of the literature suggests that daily use of either a sign language, one not based on sound, or Chinese characters, a script which maps language at the word level rather than at the phonemic level, may facilitate recall of visual forms. PMID- 14738344 TI - Relations between sport team identification and optimism. AB - Previous research indicated a positive relation between identification with a local sport team and psychological well-being. The current study was done to extend this line of research by examining the relation between identification and optimism. However, contrary to the hypothesis, the two variables were not significantly correlated for these 137 undergraduates. PMID- 14738345 TI - Sex differences in public restroom handwashing behavior associated with visual behavior prompts. AB - Handwashing after using the restroom is generally poor across the population, and one common method used to increase the frequency of handwashing is to place signs reminding individuals to wash their hands. The current study examined the association between the absence and presence of signs reminding one to wash their hands and handwashing in public restrooms. Signs prompting handwashing behavior remind restroom patrons of acceptable behavior, and the presence of these signs is hypothesized to be associated with an increase in handwashing. Observation of 175 individuals (95 women and 80 men) using public restrooms on a university campus indicated that 61% of the women and 37% of the men observed washed their hands, e.g., washing hands with soap, in the absence of the sign, and 97% of the women and 35% of the men observed washed their hands in the presence of the sign. Further, 53% of the men and 38% of the women observed rinsed their hands, e.g., washing hands without soap, in the absence of the sign, and 55% of the men and 2% of the women observed rinsed their hands in the presence of the sign. Results are discussed in terms of possible factors associated with sex differences in handwashing and the absence and presence of visual prompts for handwashing behavior. PMID- 14738346 TI - Bright light as a sleepiness prophylactic: a laboratory study of subjective ratings and EEG. AB - Sleepiness is a major problem when driving a vehicle and contributes to 15 to 30% of all road accidents. One possible countermeasure may be exposure to light. This study was designed to test whether 30 min. of exposure to a bright light would reduce subjective sleepiness and EEG indicators of sleepiness, such as alpha and theta power density. 10 female and 10 male university students (recruited through advertisements) participated in a laboratory study, consisting of 30 min. of dim light followed by 30 min. of bright light (or red light in the control condition) exposures and then by 30 min. of dim light exposure. In the dim light exposure, the luminance was 20 cd/m2, and the illuminance was about 5 Lux. In the bright exposure, the luminance was 500 cd/m2 and the illuminance about 2000 Lux. In the red light exposure, the luminance was 10 cd/m2 and the illuminance about 30 Lux. The subject sat in a chair with a seating comfort corresponding to that of a driver's seat. Analysis showed that the subjects became subjectively sleepier during the pre-exposure condition, and alpha and theta power density increased in their EEGs. The intervention significantly reduced subjective sleepiness but did not have significant effects on alpha or theta power density. The results suggest that a short (30 min.) exposure to bright light may not be a useful prophylactic against sleepiness for the period following the exposure. PMID- 14738347 TI - Effects of haptic feedback and turbulence on landing performance using an immersive cave automatic virtual environment (CAVE). AB - An investigation was conducted in which subjects had to land a simulated F-16 aircraft using a CAVE (Cave Automatic Virtual Environment) facility. This was a three-dimensional virtual setting consisting of multiple mirrors, 3-D video projected displays in a highly stressful environment employing a haptic joystick. 6 subjects learned a task which required landing in wind turbulence with a reduced visual scene. Analyses indicated that during landing, performance error variables which occurred in the same direction as the haptic forces were significantly reduced. This was true, especially when the visual scene was occluded and more reliance on the proprioceptive condition was beneficial. PMID- 14738348 TI - Neural mechanism for noise exclusion in spatial cueing. AB - Spatial cueing in an orientation discrimination task with targets embedded in high or low external noise indicates noise exclusion as a primary mechanism for attentional modulation. To implement noise exclusion in a neural network, a new mechanism is proposed based on a dendritic computation of difference between self inhibition and lateral inhibition signals. A computer simulation illustrates that the model exhibits a strong cueing effect for high noise input and no effect when the noiseless input is presented, as is consistent with behavioral signatures of noise exclusion. It is argued that the model could also exhibit object-based selection if uniform activity distribution is assumed for all cells representing the object. PMID- 14738349 TI - Ratings of contracting infectious diseases and of the risk of sexual HIV transmission by health care students from Germany, Lithuania, and Namibia. AB - In Germany, Lithuania, and Namibia different types of the HIV epidemic are present. HIV testing of health care students and their ratings of the risk of sexually acquired HIV infections and factors influencing the risk of contracting infectious diseases were analyzed. 182 German medical and dental students, 176 Lithuanian medical students, and 135 Namibian student nurses participated by completing anonymous questionnaires. The Namibian student nurses rated sexual HIV transmission risks higher than the Lithuanian and German medical students did. In general, the Lithuanian medical students rated risks of HIV transmission lowest, compared to the students in the other two samples. Risk behavior and environmental factors were most emphasized by the three student samples as having an influence on the risk of contracting infectious diseases. Thus, national prevalence of HIV influences ratings of sexual HIV transmission risks, while rating of factors influencing the risk of contracting infectious diseases are similar. PMID- 14738350 TI - Asymmetric activation in the prefrontal cortex by sound-induced affect. AB - This study is based on previous information regarding asymmetric activation in the prefrontal cortex by film-induced affects, as well as the inverse proportionality of prefrontal cortex activity to power in the alpha band of EEG. To search for a specific EEG band where the asymmetric activation in the prefrontal cortex by sound-induced affects is mainly reflected, we measured 32 college students' EEGs; 11 bands ranged from 6.5 to 35.0 Hz, at Fp1 and Fp2 sites. The power in the alpha band (8.0 to 13.0 Hz) at Fp2, especially in the alpha-2 band (9.0 to 11.0 Hz) increased while the students listened to music, during which participants reported positive affect. In contrast, the power at Fp1 increased while the students listened to noise, during which participants reported negative affect. These results imply that sound-induced positive affect increases relative left-sided activation in the prefrontal cortex, whereas induced negative affect elicits the opposite pattern of asymmetric activation. PMID- 14738351 TI - Screening for depression in clinical practice: reliability and validity of a five item subset of the CES-Depression. AB - Individuals with chronic disease are not screened routinely for depression. Availability of an abbreviated test with demonstrated reliability and validity might encourage screening so we explored the reliability and validity of a 5-item subset of the 20-item Center for Epidemiological Studies Depression Scale among inner-city outpatients with chronic asthma or diabetes. Most patients were female (73.1%) and Hispanic (61.8%). Acceptable reliability was shown by Cronbach alpha (.76) for the subset of 5 items. Validity was supported by the high correlation of .91 between patients' scores on the 5-item subset and the full 20 items. The 5 items reflected a single factor (eigenvalue = 2.66). Receiver operating characteristic curve analysis identified cut-points for the 5 items that were sensitive (> .84) and specific (> or = .80) in identifying patients classified as depressed by full 20 items. The reduced patient and clinician burden of the subset of 5 items, as well as its desirable psychometric properties, support broader application of this subset as a screening tool for depression. PMID- 14738352 TI - Self-perceived and actual indicators of motor abilities in children and adolescents. AB - The aim of the study was to estimate the correlations between self-perceived and actual indicators of fitness in children and adolescents of different age and sex. A total of 525 boys and girls, ages 10 to 17 years (divided into the four age groups) were studied. Self-perceived fitness (fitness, endurance, strength, flexibility, and body composition) was assessed using a slightly modified version of a questionnaire by Delignieres, Marcellini, Brisswalter, and Legros, which was described by Lamb and Haworth. The following EUROFIT tests were used: 20-m endurance shuttle-run, hand-grip dynamometry, and sit-and-reach. Nine skinfolds were summarized as a body-composition parameter. The subjects significantly perceived their endurance, flexibility (except 10- to 11-yr.-olds), and body composition (except 10- to 11-yr.-old boys). Correlations between handgrip dynamometry and perceived strength were not significant. PMID- 14738353 TI - Upper and lower body strength in relation to ball speed during a serve by male collegiate tennis players. AB - The relation of leg, shoulder, and grip strength to ball speed in the tennis serve was investigated. For 15 collegiate male tennis players, leg and shoulder strength were measured using a Lido Active isokinetic dynamometer, grip strength with a handgrip dynamometer, and ball speed with a radar gun. Regression analysis showed no significant (p<.05) relationship among the strength variables and ball speed. Results were explained in terms of strength not being the only factor involved in producing ball speed during the tennis serve. PMID- 14738354 TI - A study of field independence versus field dependence of school teachers and university students in mathematics. AB - The Group Embedded Figures Test was administered to 72 secondary school teachers and 54 university students in mathematics to measure their cognitive styles of field independence-dependence. A mean difference was found between the teachers and students as teachers scored more field-independent than the students. There was also a group-by-sex interaction, which indicated that the female teachers scored more field-independent than the male teachers, whereas the male students scored as more field-independent than the female students. Implications of the findings are reflected in the discussion. PMID- 14738355 TI - Enhanced power within a specific band of theta activity in one person while another receives circumcerebral pulsed magnetic fields: a mechanism for cognitive influence at a distance? AB - Four pairs of adult siblings served once as either the stimulus or the response person in two sessions separated by one week. While the brain of the stimulus person, who was seated in a closed chamber, was exposed successively to six different complex magnetic fields for 5 min. each quantitative monopolar electroencephalographic measurements over the frontal, temporal, parietal, and occipital lobes were collected by computer for the response person who was seated in another room. The six configurations of fields served as different stimulus patterns and had been designed to affect consciousness. As predicted theoretically, a significant increase in electroencephalographic power within the 5.0-Hz to 6.0-Hz band over the frontal and occipital lobes was noted for the response persons when the stimulus persons received one of the six specific patterns of weak (1 microTesla range) magnetic fields. This magnetic stimulus was presented for 100 msec. with changes in rate of 20 msec. to each of the eight solenoids that were equally spaced in the horizontal plane around the head of the stimulus person. Derivatives within this narrow frequency band had been hypothesized to be a source of the "binding factor" for the cohesive cerebrogenic electromagnetic fields producing consciousness. The results suggest that an appropriate altered state of one brain can effect specific predictable frequencies of the electroncephalographic activity of another distant brain which is genetically related. PMID- 14738356 TI - Somatosensory event-related potentials (ERPs) associated with stopping ongoing movement. AB - The somatosensory event-related potentials (ERPs) associated with stopping ongoing movement and increasing muscular tension were examined. 14 healthy right handed volunteers, 10 men and 4 women (21-29 years old, M age +/- SD, 24.1 +/- 2.5 yr.) performed a stop/increase reaction task. They were requested to perform an elbow extension movement with the right arm and to maintain 20% of the maximum voluntary contraction forces (MVC) before the electrical stimuli were delivered to either the left index finger or the left little finger. They executed one of two movements from the sustained contraction state: they had to stop the muscular tension following the left little finger stimulus or increase the muscular tension from 20% to 40% of the maximum voluntary contraction forces following the left index finger stimulus. The reaction time and somatosensory sequence P100 N140-P300 components of event-related potentials were recorded for each electrical stimulus, respectively. The reaction time was longer to the increase reaction condition than to the stop reaction condition. Neither P100 nor N140 components showed significant differences between stop and increase reaction conditions. The P300 to the stop reaction condition was of greater amplitude and latency than those of the increase reaction condition. These results suggest that stopping the ongoing movement processing requires a longer stimulus evaluation time and is more demanding than increasing reaction processing. PMID- 14738357 TI - On the relation between power and reliability of difference scores. AB - The potential problems which may arise from the use of difference (a.k.a., gain) scores in the measurement of change are well documented, including (a) difference scores are often negatively correlated with initial ability, and (b) they often tend to be unreliable. Conversely, they make excellent dependent variables in a true experiment since they tend to reduce variability due to individual differences among persons. This brief didactic paper presents a conclusion similar to Nicewander and Price, using a perhaps more straightforward argument based on difference scores. We argue that the same reason difference scores provide powerful significance tests, namely, reduction of "true score" variance, is also the reason they tend to be unreliable. Further, we make the point that reducing true score variance will increase the power of a significance test (since it will reduce the denominator or "error term" of the observed statistic) but will decrease reliability (since it is the numerator and a component of the denominator of the reliability coefficient). PMID- 14738358 TI - Using microswitches with persons who have profound multiple disabilities: evaluation of three cases. AB - This study evaluated the effectiveness of microswitch programs in promoting and maintaining responding of persons with profound multiple disabilities. Three participants were provided one or two microswitches. Their responding (microswitch activation) produced favorite stimuli. Analysis showed that all participants increased responding significantly, albeit with clear differences among them, during the intervention period of a few weeks. Two participants continued with the microswitch program in the daily context and had 3-mo. follow up data similar to the intervention data. One participant did not continue with the program (reportedly because of inconsistent/low responding) and so did not receive a follow-up assessment. Links between responding and vigilance conditions and stereotypy are discussed. PMID- 14738359 TI - Quantification of human spatial behavior in an open field-locomotor maze. AB - To obtain and analyze the strategies of human spatial behavior, the Kiel Locomotor Maze, a maze-like analogue, was used. The Kiel Locomotor Maze automatically records different types of spatial memory errors, distances and rotation angles, decision time, and reaction times for each move. 18 patients with cerebral tumors within the frontal, temporal, or parietal lobes and 16 patients with Parkinson's disease were investigated with respect to "sense of direction." These parameters provide information beyond error scores. It was expected that patients with focal cortical lesions would be characterised by loss of directional sense whereas basal ganglia-related brain disease is characterised by preserved directional sense but poor memory for cue-defined target locations. Based on the neuropsychological theory of head-direction sense, especially the 1991 model of McNaughton, the functionality of storage and update of the head direction sense may be reflected with the help of these parameters. "Direction sense" parameters provide information about success or failure of the acquisition of a spatial task much earlier than error scores. PMID- 14738360 TI - Role of movement amplitude to timing control in bimanual coordination. AB - A bimanual coordinative pattern consists of the movement frequency, movement amplitude, and relative phase between the two hands. This study examined a role of the movement amplitude to timing in the bimanual coordination. 12 subjects (8 men and 4 women; mean age of 21.8 yr.) reproduced 1:1, 2:1, 3:1, 3:2, and 5:3 polyrhythmic tapping patterns. No difference was shown between the amplitudes of the two hands in each pattern. However, the amplitude of complex (3:2 and 5:3) patterns was larger than of simple (1:1, 2:1, and 3:1) patterns. It is concluded that the complicated timing control is attained by increasing the amplitude. PMID- 14738361 TI - Relations of women exercisers' mastery and performance goals to traits, fitness, and preferred styles of instructors. AB - This study addressed the influence of both mastery and performance goals of 144 women (M=21.8 yr.) exercising at a campus fitness center. Five goal patterns were compared: mastery alone, performance alone, both-low, mastery-performance, and, performance-mastery. These five patterns were compared in terms of optimism, social physique anxiety, and fitness as well as preferences for instructors' mastery, performance, or supportive responses. Those with dual motivational goals (mastery-performance and performance-mastery) preferred instructors to emphasize mastery and supportive responses, and they had higher fitness than the mastery or low groups. However, the performance-mastery group endorsed significantly lower optimism than all other groups. There were no group differences on the measure of anxiety about physique. Results support the importance of addressing both mastery and performance goals when seeking to understand and influence the attitudes of women who work out in a fitness center. Instructors would do best to emphasize autonomy and supportiveness and avoid comparisons among women exercisers. PMID- 14738362 TI - Increased proportions of sensed presences and occipital spikes with 1- and 10 msec. point duration of continuous 7-Hz transcerebral magnetic fields. AB - To test the hypothesis that the numbers of "sensed presences" induced by the application of weak (1 microTesla) magnetic fields over the right hemisphere was subject to an optimal time of presentation, digital-to-analogue computer generated 7-Hz sine-waves were applied with point ("pixel") durations of 1, 3, 5, 7, 10, or 20 msec. for 5 min. each in a sequential counterbalanced order for six subjects. The proportion of numbers of sensed presences reported along the subjects' left sides and the proportions of electroencephalographic spikes over the occipital regions were greatest when 1- or 10-msec. point durations were employed. These results indicate that the specific duration of each successive field strength composing a complex sequence should be considered when designing optimal configurations for intercalating with neurocognition. PMID- 14738363 TI - Sex-typing and prestige ratings of occupations as indices of occupational stereotypes. AB - In the present study, an ethnically diverse convenience sample (N=295; 56% female) of working adults (47%) and college students (age M=23.7, SD=6.7) rated a list of 25 occupations on a 9-point sex-typing scale, with anchors of 1: mostly associated with men and 9: mostly associated with women. They also rated the status or prestige of these occupations on an 11-point Prestige scale, with anchors of 1: low prestige/status and 11: high prestige/status. Findings were consistent with results reported by Panek, Rush, and Greenawalt in 1977 and O'Connor in 1982. Little has changed: respondents still hold sex-based opinions about occupations. Moreover, occupational titles perceived as mostly associated with men were rated higher in prestige than occupational titles perceived as mostly associated with women. This pattern was found for both high and low prestige jobs. Men and women did not differ in their evaluations of these occupations. PMID- 14738364 TI - Effect of teaching with mirrors on ballet dance performance. AB - This study assessed the effect of mirrors on the dance performance of beginning college ballet students in the classroom setting, using an evaluation methodology developed for this study. 13 women enrolled in one ballet class were taught using mirrors, 14 women in a second beginning ballet class were taught without mirrors. Both classes were taught by the same instructor. All students were videotaped performing the same adagio and grand allegro phrase during Weeks 5 and 14 of the 14-wk. term. At the end of the semester two ballet teachers viewed the videotapes for both classes. One evaluator was the instructor, and the other was a blind reviewer who had no knowledge of which was the mirror and nonmirror class during the evaluation process. They were instructed to choose a score for each dancer with anchors of 1: low skill and 5: high skill for both the adagio and allegro phrases. High interrater reliability was noted for both the adagio and allegro phrase data. In the nonmirror class, there was a significant increase in adagio scores, but no significant increases in adagio and allegro scores for the mirror class. These results suggest that the use of the mirror in a ballet classroom may negatively affect skill acquisition of the dancer. PMID- 14738365 TI - Generation and sex differences in sensation seeking: results of the family study. AB - Previous research regarding sex and age differences for sensation seeking in English, American, Canadian, and Australian samples showed clear sex differences and a decline of sensation seeking across ages. The 1978 Form V of the Sensation Seeking Scale was administered to a Croatian sample of 151 high school students and their 226 parents. Analysis showed that parents had lower mean scores than their children, consistent with the hypothesis that sensation seeking declines with age. Male subjects scored higher on the total Sensation Seeking, and Thrill and Adventure Seeking, Disinhibition, and Boredom Susceptibility subscales than female subjects, while there was no difference between male and female subjects on the Experience Seeking scale. Also, several sex-by-generation interactions were significant. PMID- 14738366 TI - Perceptual defense in anxiety disorders. AB - Prior research by MacLeod and Rutherford (1992) indicates that anxious subjects could have perceptual strategies different from nonanxious subjects. 42 verbal stimuli of six types (disease, social anxiety, panic, agoraphobia, obsessive compulsive, and neutral) were tachistoscopically presented to three groups of subjects, aged 18 to 60 years: Panic Disorder group (n=21: 13 women and 8 men), and Obsessive-Compulsive Disorder group (n=20: 14 women and 6 men), recruited from an outpatient clinic, and a Control group (n=22: 14 women and 8 men), recruited among students and hospital staff. The times required for correct identification were generally longer for anxious subjects but quicker for stimuli specifically related to their disorder. The data could indicate a two-step perceptual strategy or two distinct ways of perceiving, usually, a generalized perceptual defense for a majority of anxiety-loaded stimuli, but also a selectively facilitated processing for stimuli specific to the disorder. PMID- 14738367 TI - Qualitative analysis of the overhand throw by undergraduates in education using a distance learning computer program. AB - The purpose was to (a) examine whether computer-based distance learning could enhance the qualitative analysis skills (error detection in the overhand throw) of undergraduates in education and (b) examine the effectiveness of several methods of information presentation (video file and text) on distance learning. Participants were randomly assigned to 1 of 4 groups, to detect errors in an incorrect throwing motion of a model on the computer screen. Group 1 (n=13) was the control; Group 2 (n=13) viewed a video of the appropriate throwing mechanics; Group 3 (n=13) viewed text information describing the appropriate mechanics of the overhand throw; and Group 4 (n=16) received a combination of video and text information. On Day 1 participants took a pretest. Treatment and testing occurred on Days 2 through 8. Then 5 days later participants took a retention test. One way analysis of variance confirmed no significant differences between groups at Pretest (Day 1). An analysis of variance with repeated measures indicated learning over practice. Paired-sample t tests between Days 1 and 8 showed the video plus text group without significant change. PMID- 14738368 TI - Distortion-product otoacoustic emissions in ears with normal hearing sensitivity: test-retest variability. AB - 4 women and 4 men with normal hearing sensitivity were tested for variability in distortion-product otoacoustic emissions, with two measurement sessions per day for 4 days spread over a 2-wk. period. Most test-retest variations were small, on the order of 2 dB. Extreme variations of 6 dB or greater occurred most often at the highest test frequency and should not be considered clinically positive unless such differences are found at lower frequencies also. PMID- 14738369 TI - The PsycARTICLES database: attributes and limitations. AB - This paper presents an overview on the main features of the new APA database, PsycARTICLES, with a focus on the citation searching option. Both positive aspects and limitations of the file are noted. PMID- 14738370 TI - Motivation, stress, and cortisol responses in skydiving. AB - This study examined metamotivational dominance, metamotivational states, and cortisol responses in skydiving participants. Data were obtained from 23 experienced skydivers 15 min. prior to and following a skydive. Respondents were mainly paratelic-conformist dominant and most occupied the conformist and arousal seeking states prior to skydiving, assessed respectively, with Apter, et al.'s Motivational Style Profile and Cook, et al.'s measure of metamotivational states. Paratelic-conformist dominance indicates a predisposition towards conformity and a desire to be spontaneous, and the conformist and arousal seeking states reported prior to completing the skydive represent a desire to conform to expected norms but also to seek arousal. There was no significant change in scores for metamotivational state or stress following skydiving. Contrary to expectations, cortisol level prior to skydiving was negatively associated with external stress. These results support the paratelic, but not the negativistic, dominance found in previous samples of risk sport participants (no skydivers). The conformist dominance and pre-dive conformist metamotivational state scores of this sample may be fundamental for adhering to safety regulations imposed on skydivers. To obtain better understanding of this phenomenon, researchers should attempt to measure these variables during, rather than prior to and after, participation in risk sports. PMID- 14738371 TI - Will safety release cords on children's cameras prevent strangulation? AB - This study investigated the automatic safety release mechanisms on neck cords for children's cameras. The purpose of the safety release mechanism is to reduce accidental strangulation associated with children's toys that have neck cords. The horizontal pull forces that activate the automatic safety release mechanism were measured for two different styles of release mechanisms for neck cords. When these forces were compared to forces associated with child strangulation, the automatic safety mechanisms released at higher forces than the forces associated with accidental strangulation or death, which is contrary to what was intended and creates a false sense of security for parents who select a toy camera with such a device. PMID- 14738372 TI - Improved performance on clerical tasks associated with administration of peppermint odor. AB - Previous research indicates the presence of certain odors is associated with enhanced task performance. The present study investigated use of peppermint odor during typing performance, memorization, and alphabetization. Participants completed the protocol twice--once with peppermint odor present and once without. Analysis indicated significant differences in the gross speed, net speed, and accuracy on the typing task, with odor associated with improved performance. Alphabetization also improved significantly under the odor condition but not typing duration or memorization. These results suggest peppermint odor may promote a general arousal of attention, so participants stay focused on their task and increase performance. PMID- 14738373 TI - The cancer/testis genes: review, standardization, and commentary. AB - Cancer/testis (CT) antigens are immunogenic in cancer patients, exhibit highly tissue-restricted expression, and are considered promising target molecules for cancer vaccines. To date, 44 CT gene families have been identified and their expression studied in numerous cancer types. For example, bladder cancer, non small cell lung cancer, and melanoma are high CT gene expressors, with 11/20 (55%), 17/33 (51%) and 17/32 (53%) of the CT transcripts examined by RT-PCR detected in 20% or more of the specimens examined, respectively. Breast and prostate cancer can be considered moderate CT gene expressors, with 12/32 (37%) and 6/20 (30%) CT transcripts having an expression frequency >20%, respectively, while renal and colon cancer are low CT gene expressors, with only 3/33 (9%) and 4/25 (16%) CT transcripts having an expression frequency >20%, respectively. In normal tissues, standardized RT-PCR experiments showed that 19/43 CT genes were testis-restricted, 10/43 CT genes were tissue-restricted (mRNA detected in 2 or fewer non-gametogenic tissues), 9/43 CT genes were differentially expressed (mRNA detected in 3-6 non-gametogenic tissues), and 5/43 CT genes were ubiquitously expressed. With the exception of testis-restricted CT transcripts, all remaining CT transcripts were expressed in normal pancreas. In terms of immunogenicity, 14/29 testis/tissue-restricted CT gene families have been shown to induce a cellular and/or humoral immune response in humans. In view of the expanding list of CT genes, a CT gene database was created to standardize CT nomenclature and accumulate relevant data regarding their expression profiles, immunogenicity, function (where known), gene structure and location, and orthologous groups. PMID- 14738375 TI - Ponicidin and oridonin are responsible for the antiangiogenic activity of Rabdosia rubescens, a constituent of the herbal supplement PC SPES. AB - Antiangiogenic activity has been identified in an aqueous EtOH extract of Rabdosia rubescens, a component of the dietary supplement PC SPES. Bioassay guided fractionation using a novel in vitro human endothelial cell-based assay for angiogenesis afforded the diterpenoids ponicidin (1) and oridonin (2), with significant antiangiogenic activity at subcytotoxic concentrations, suggesting that these constituents may strongly contribute to the demonstrated clinical efficacy of PC SPES as a treatment for advanced prostate cancer. PMID- 14738376 TI - Spirostanol and furostanol glycosides from the fresh tubers of Polianthes tuberosa. AB - Six new steroid glycosides--two spirostanols, polianthosides B and C (1, 2), and four furostanols, polianthosides D-G (3-6)--were isolated from the fresh tubers of Polianthes tuberosa, together with seven known spirostanols (7-13) and a known furostanol (14) saponins. Their structures were elucidated on the basis of spectroscopic analysis and the results of acidic and enzymatic hydrolysis. The cytotoxic activities of 1-14 against HeLa cells are reported. PMID- 14738377 TI - Saponins from the flower buds of Buddleja officinalis. AB - Five new saponins, mimengosides C-G (1-5), were isolated from the flower buds of Buddleja officinalis along with five known compounds, namely, songaroside A, acteoside, phenylethyl 2-glucoside, echinacoside, and phenylethyl alcohol 8-O beta-D-glucopyranosyl-(1-->2)-beta-D-glucopyranoside. The structures of 1-5 were elucidated using spectroscopic and chemical methods, and these compounds were evaluated for their inhibitory effects against HL-60 leukemia cells. PMID- 14738378 TI - Insecticidal sesquiterpene pyridine alkaloids from Maytenus chiapensis. AB - The new sesquiterpene pyridine alkaloids chiapenines ES-I (1), ES-II (2), ES-III (3), and ES-IV (4), in addition to the known alkaloids wilfordine (5), alatamine (6), wilforidine (7), alatusinine (8), euonine (9), euonymine (10), ebenifoline E I (11), forrestine (12), mayteine (13), and 4-hydroxy-7-epi-chuchuhuanine E-V (14), were isolated from the leaves of Maytenus chiapensis. Their structures were elucidated by 1D and 2D NMR spectroscopy, including homonuclear and heteronuclear correlation (COSY, ROESY, HSQC, and HMBC) experiments. Wilfordine, alatusinine, and euonine exhibited strong antifeedant activity against Spodoptera littoralis. PMID- 14738379 TI - Antifungal 3-butylisocoumarins from Asteraceae-Anthemideae. AB - Seven new naturally occurring 3-butylisocoumarins were isolated and identified from lipophilic extracts of aerial as well as underground organs: corfin (17) and 3'-hydroxycorfin (18) from the roots of Chamaemelum mixtum and (-)-(R)-2' methoxydihydroartemidin (5), (+)-(S,R)-epoxyartemidin (6a), dracumerin (12), (+) (R)-(E)-3'-hydroxyartemidin (13), and capillarin isovalerate (20) from various organs of Artemisia dracunculus (tarragon). Furthermore, six known derivatives, artemidiol (7), (E/Z)-artemidin (11), capillarin (19), artemidinol (21), 8 hydroxyartemidin (22), and 8-hydroxycapillarin (23), were obtained. The antifungal activities of all naturally occurring derivatives were determined in a germ-tube inhibition test against a susceptible strain of rice blast fungus Pyricularia grisea. The 3-butyl side-chain is a prerequisite for high activity. Eleven structurally related synthetic derivatives were additionally tested to explore the influence of structural characteristics on activity. Benlate, blasticidin S, kresoxim-methyl, griseofulvin, and the carrot phytoalexin 6 methoxymellein all served as positive controls. PMID- 14738380 TI - New geranyl stilbenes from Dalea purpurea with in vitro opioid receptor affinity. AB - Three new geranyl stilbenes, pawhuskins A, B, and C (1, 2, and 3), were isolated from organic extracts of Dalea purpurea. The structures of the three compounds were determined by NMR and HRMS methods. The activities of these compounds, along with that of the known compound petalostemumol (4), were evaluated in an opioid receptor assay in vitro. Pawhuskin A (1) exhibited the strongest activity of the four compounds with a K(i) value of 0.29 +/- 0.11 microM. PMID- 14738381 TI - Africane-type sesquiterpenoids from the Argentine liverwort Porella swartziana and their antibacterial activity. AB - Seven africanes (1, 2a,b, 3-6), two of them new (1, 2a), three secoafricanes (7 9), one of them new (7), and two norsecoafricanes (10, 11a), one of them new (10), all of them swartzianin-type, have been isolated from an Argentine collection of the endemic liverwort Porella swartziana. The structures of the new compounds were established by extensive 1D and 2D NMR spectroscopic data. Absolute configurations of compounds 2a, 2b, and 10 were derived on the basis of CD spectra. The compounds were tested for activity against a variety of microbes, but none were found to exhibit significant antibacterial activity. PMID- 14738382 TI - Polyoxygenated eudesmanes and trans-chrysanthemanes from the aerial parts of Santolina insularis. AB - The eudesmane sesquiterpenoids 1-3 and the trans-chrysanthemyl monoterpenoid 4 have been isolated from the aerial parts of Santolina insularis, a bush endemic to Sardinia. The absolute stereostructures of these novel compounds and of two known but incompletely characterized chrysanthemanes (5, 6) were established by spectroscopic techniques and by application of the modified Mosher method. The presence of the p-menthane aldehyde eucamalol (7) gives credit to the widespread use of S. insularis to fend off mosquitoes. PMID- 14738383 TI - Tetrahydrofuran lignans from Nectandra megapotamica with trypanocidal activity. AB - The phytochemical investigation of the crude ethanol extract of the leaves of Nectandra megapotamica afforded three new (2d, 3b, and 3c) and eight known (1a-d, 2a-c, and 3a) tetrahydrofuran lignans. Regarding the in vitro trypanocidal activity, the lignan-rich fraction, at a concentration of 2.0 mg/mL, was 100% active. Compound 1a was the most active, showing an IC(50) value of 2.2 microM and lysis of 94% of the parasites at 32 microM. Compounds 1b, 1d, 2a, 2c, and 3a displayed moderate activity, while compound 1c was inactive. PMID- 14738384 TI - Five new maleic and succinic acid derivatives from the mycelium of Antrodia camphorata and their cytotoxic effects on LLC tumor cell line. AB - Five new maleic and succinic acid derivatives were isolated from the mycelium of Antrodia camphorata. Their structures were determined by various spectroscopic means. Maleimide derivatives 2 and 3 showed appreciable cytotoxic activity against LLC cells. PMID- 14738385 TI - Micromide and guamamide: cytotoxic alkaloids from a species of the marine cyanobacterium Symploca. AB - Two new cytotoxins have been isolated from a species of marine cyanobacterium belonging to the genus Symploca that was collected in Guam. These new compounds, micromide (1) and guamamide (2), were accompanied by the known lipopeptides apramides A (3), B (4), and G (5). The planar structures of both alkaloids were elucidated by standard 2D NMR techniques, and the configurations of the amino acid-derived units in 1 were determined by chiral HPLC. The stereochemistry of the beta-methoxyhexanoic acid in 1 was determined by derivatization with methyl d mandelate, after acid hydrolysis, and comparison with synthetic standards. PMID- 14738386 TI - New alkaloids from Pandanus amaryllifolius. AB - Three new alkaloids, the two pyrrolidine type alkaloids (1 and 2) and 6E pandanamine (3), together with five known alkaloids (4-8), were isolated from the leaves of Pandanus amaryllifolius collected in West Java, Indonesia. All the new alkaloids have two alpha-methyl alpha,beta-unsaturated gamma-lactone moieties, while compound 2 also has an additional seven-membered ring, which has not been encountered before in Pandanus alkaloids. Two different extraction methods, namely, a solvent partitioning extraction and acid-base treatment, were tested, giving secondary and tertiary amines, respectively. Spectroscopic and chemical studies showed that the tertiary amines isolated from the acid-base treatment were artifacts formed during the extraction process. This finding suggests that the use of conventional acid-base treatment in isolating Pandanus alkaloids should be reviewed since it can introduce artifacts. PMID- 14738387 TI - Production of biologically active taxoids by a callus culture of Taxus cuspidata. AB - Ten known taxoids, paclitaxel, 7-epi-taxol, taxol C, baccatin VI, taxayuntin C, taxuyunnanine C and its analogues (2-5), and yunnanxane (6), and an abietane, taxamairin A, were produced in the callus culture of Taxus cuspidata cultivated on a modified Gamborg's B5 medium in the presence of 0.5 mg/L NAA. After stimulation with 100 microM methyl jasmonate, five more taxoids, cephalomannine, 1beta-dehydroxybaccatin VI, taxinine NN-11 (1), baccatin I, and 2alpha acetoxytaxusin, and one more abietane, taxamairin C, were found in addition to the above-mentioned compounds. It was also observed that the content of the products increased over three times. Taxinine NN-11 (1) is a new taxane whose structure was assigned as 5alpha,13alpha-diacetoxy-9alpha-cinnamoyloxy-4(20),11 taxadien-10beta-ol by analysis of its spectral data. Taxinine NN-11 (1) exhibited significant MDR reversal activity toward 2780 AD tumor cells. The results of primary screening based on 39 human cancer cell lines suggest that 1 also belongs to a new mechanistic class. Efficient production of 1 was investigated using the callus culture of T. cuspidata. PMID- 14738388 TI - The microbiological transformation of two 15beta-hydroxy-ent-kaurene diterpenes by Gibberella fujikuroi. AB - The incubation of 15beta-hydroxy-3-oxo-ent-kaur-16-ene (1) with the fungus Gibberella fujikuroi afforded 11beta-hydroxy-3,15-dioxo-ent-kaurane (6), 11beta,15beta-dihydroxy-3-oxo-ent-kaur-16-ene (8), 7beta,11beta,15beta-trihydroxy 3-oxo-ent-kaur-16-ene (9), 7alpha,11beta-dihydroxy-3,15-dioxo-ent-kaurane (7), and 7alpha,11beta,15beta-trihydroxy-3-oxo-ent-kaur-16-ene (10). The incubation of 15beta-hydroxy-ent-kaur-2,16-diene (3) with the same fungus yielded 7alpha,11beta dihydroxy-15-oxo-ent-kaur-2-ene (12), 7alpha,11beta,15beta-trihydroxy-ent-kaur 2,16-diene (13), 7beta,15beta-dihydroxy-ent-kaur-2,16-dien-19,6-olide (14), 1beta,7beta,15beta-trihydroxy-ent-kaur-2,16-dien-19-oic acid (15), 7alpha,11beta,16alpha-trihydroxy-15-oxo-ent-kaur-2-ene (17), and 7alpha,15beta,17 trihydroxy-11beta,16beta-epoxy-ent-kaur-2-ene (19). These results indicated that a 3-oxo group in ent-kaur-16-ene derivatives inhibits the oxidation at C-19, typical of the biosynthetic pathway of gibberellins and kaurenolides, while a 2,3 double bond or a 15beta-OH does not. In both substrates a 15beta-alcohol directs hydroxylations at C-11(beta) and C-7(alpha), while in those with a 2,3-double bond the functionalization of C-1(beta) is favored. PMID- 14738389 TI - Caulibugulones A-F, novel cytotoxic isoquinoline quinones and iminoquinones from the marine bryozoan Caulibugula intermis. AB - An extract of the marine bryozoan Caulibugula intermis, collected in the Indo Pacific off Palau, produced a distinct pattern of differential cytotoxicity in the National Cancer Institute's 60 cell line antitumor screen. Bioactivity directed fractionation of the extract provided six new compounds, caulibugulones A-F (1-6). The structures of these novel metabolites were determined by spectrochemical analyses including LC-MS, HRFABMS, 1-D and 2-D NMR experiments, and by comparison with related compounds. The structures of compounds 2 and 3 were confirmed by chemical interconversion. The isolated compounds exhibited IC(50)'s of 0.03-1.67 microg/mL against murine tumor cells in an in vitro cytotoxicity assay. PMID- 14738390 TI - Vibsane diterpenoids from the leaves and flowers of Viburnum odoratissimum. AB - In addition to the five known compounds 5-epi-vibsanin H, vibsanins C, H, and G, and aldovibsanin B, four new diterpenes, 5-epi-vibsanin G (1), 18-O methylvibsanin G (2), vibsanin M (3), and aldovibsanin C (4), were isolated from an acetone extract of the leaves and flowers of Viburnum odoratissimum by bioassay-directed fractionation. In addition, two acetyl derivatives 5 and 6 were obtained from the naturally occurring diterpenes. The structures of the new compounds were established on the basis of NMR spectral analysis, including COSY, HMQC, HMBC, and NOESY correlations. The compounds were evaluated for cytotoxicity against human nasopharyngeal carcinoma (HONE-1) tumor cells and human gastric cancer (NUGC-3) cells. PMID- 14738391 TI - New communesin derivatives from the fungus Penicillium sp. derived from the Mediterranean sponge Axinella verrucosa. AB - The ethyl acetate extract of Penicillium sp., derived from the Mediterranean sponge Axinella verrucosa, yielded the known compound communesin B (1) and its new congeners communesins C (2) and D (3), as well as the known compounds griseofulvin, dechlorogriseofulvin, and oxaline. All structures were unambiguously established by 1D and 2D NMR and MS data. In several bioassays performed on different leukemia cell lines, the communesins exhibited moderate antiproliferative activity. PMID- 14738392 TI - A pair of new C-21 steroidal glycoside epimers from the roots of Cynanchum paniculatum. AB - Two new C-21 steroidal glycoside epimers, paniculatumosides A and B (1 and 2), based on a 13,14:14,15-disecopregnane-type skeleton, with neocynapanogenin B (3) and neocynapanogenin C (4) as two new aglycons, were isolated from the roots of Cynanchum paniculatum. Their structures were determined by spectral data interpretation. PMID- 14738393 TI - Two new analogues of the cytotoxic substance BE-52211 from Streptomyces sp. AB - Two new beta-hydroxy acetamides, BE-52211 B and BE-52211 C, which were structural analogues of BE-52211, were obtained as an inseparable mixture from an actinomycete, Streptomyces sp. Their structures were elucidated on the basis of spectroscopic data. They inhibited cell division of starfish embryos at a concentration of 2.5 microg/mL or greater. PMID- 14738394 TI - Syncarpamide, a new antiplasmodial (+)-norepinephrine derivative from Zanthoxylum syncarpum. AB - A new (+)-norepinephrine derivative, syncarpamide (1), along with a known coumarin, (+)-S-marmesin (2), and one known alkaloid, decarine (3), have been isolated from the stem of Zanthoxylum syncarpum. The structure of compound 1 was elucidated on the basis of 1D and 2D NMR, MS, IR, optical rotation, and CD analyses. Its absolute stereochemistry was elucidated by synthesis of its enantiomer and subsequent comparison of CD data. Characterizations of compounds 2 and 3 were based on spectral analysis and comparison with reported data. Compounds 1 and 3 showed antiplasmodial activity, with IC(50) values of 2.04 and 1.44 microM against Plasmodium falciparum D(6) clone and 3.06 and 0.88 microM against P. falciparum W(2) clone, respectively. Compound 3 showed cytotoxicity at 56.42 microM, whereas compound 1 was not cytotoxic at 10.42 microM. Compound 1 was tested for hypotensive activity, but no activity was observed. Compound 2 showed no antiplasmodial or antimicrobial activities. PMID- 14738395 TI - Three new oleanane-type triterpenes from Ludwigia octovalvis with cytotoxic activity against two human cancer cell lines. AB - Three new oleanane-type triterpenes, (23Z)-coumaroylhederagenin (1), (23E) coumaroylhederagenin (2), and (3Z)-coumaroylhederagenin (3), together with two known triterpene acids, oleanolic acid and ursolic acid, have been isolated from the whole plant of Ludwigia octovalvis, and their structures have been elucidated by spectroscopic methods. All three new triterpenes showed significant cytotoxicity against two human tumor cell lines, namely, oral epidermoid carcinoma KB and colorectal carcinoma HT29, and gave IC(50) values in the range 1.2-3.6 microM. PMID- 14738396 TI - Lancifodilactones B-E, new nortriterpenes from Schisandra lancifolia. AB - Four new nortriterpenes, lancifodilactones B-E (1-4), were isolated from the leaves and stems of Schisandra lancifolia. Their structures were determined by analysis of 1D and 2D NMR spectroscopic data. These compounds are members of a new highly oxygenated cycloartane skeletal class with a biosynthetically modified eight-membered ring D. PMID- 14738397 TI - Immunomodulatory constituents from an Ascomycete, Chaetomium seminudum. AB - A screening study focusing on immunomodulatory activity of the EtOAc extract of an Ascomycete, Chaetomium seminudum, has afforded a known epipolythiodioxopiperazine, chetomin (1), together with three new chetomin related metabolites named chetoseminudins A (2), B (3), and C (4). Among these four metabolites, 1 and 2 have been deduced as the immunosuppressive features of this fungus. PMID- 14738398 TI - A new phloroglucinol derivative from the brown alga Eisenia bicyclis: potential for the effective treatment of diabetic complications. AB - A new phloroglucinol derivative (1) and two known compounds (2 and 3) were isolated from the brown alga Eisenia bicyclis. These isolates exhibit inhibitory activity on glycation and alpha-amylase. Their structures were determined on the basis of spectroscopic data. PMID- 14738399 TI - Asmarines G and H and barekol, three new compounds from the marine sponge Raspailia sp. AB - Four asmarines, the known A and F and two new ones G and H, four diterpenes, the known chelodane, barekoxide, and zaatirin and a new one, barekol, and methyl 3 oxo-cholan-24-oate were isolated from the Kenyan sponge Raspailia sp. The structures of all these compounds were established on the basis of MS and NMR data, and the absolute configuration of barekol was determined from the CD measurements of its keto-derivative. The (15)N chemical shifts of the N atoms of the heterocycles of the asmarines were measured from (15)N HMBC experiments, and the influence of various structural features on the delta(N) values studied. PMID- 14738400 TI - Plakolide A, a new gamma-lactone from the marine sponge Plakortis sp. AB - Plakolide A (1), a new alpha-exomethylene-gamma-lactone isolated from the marine sponge Plakortis sp., was found to inhibit inducible nitric oxide synthase (iNOS) activity. The isolation, structure elucidation, and biological activity of plakolide A is described. PMID- 14738401 TI - Tasnemoxides A-C, new cytotoxic cyclic norsesterterpene peroxides from the Red Sea sponge Diacarnus erythraenus. AB - Tasnemoxides A-C (1-3), three new cytotoxic cyclic norsestertepene peroxides, were isolated from the Red Sea sponge Diacarnus erythraenus, together with the known compound sigmosceptrellin B (4). The structural determination of the isolated compounds was based on extensive 1D and 2D NMR studies and mass spectral determinations. Compounds 1-3 showed moderate cytotoxicity against three cancer cell lines. PMID- 14738402 TI - Structure of new carotenoids with the 6-oxo-kappa end group from the fruits of paprika, Capsicum annuum. AB - New carotenoids 1 and 2 were isolated as minor components from the ripe fruits of paprika (Capsicum annuum). The structures of 1 and 2 were determined to be (3R,5'R)-3-hydroxy-beta,kappa-caroten-6'-one and (5'R)-3,4-didehydro-beta,kappa caroten-6'-one, respectively, from UV-vis, NMR, CD, HRFABMS, and FABMS/MS spectra. PMID- 14738403 TI - Highly hydroxylated triterpenes from Salvia kronenburgii. AB - The three new triterpenes (1-3) and five known triterpenes and a sterol were isolated from the acetone extract of a Turkish collection of Salvia kronenburgii. The structures of the new triterpenes were established as 1beta,2alpha-dihydroxy 3beta-acetoxy-11-oxours-12-ene (1), 2alpha,20beta-dihydroxy-3beta-acetoxyurs 9(11),12-diene (2), and 1beta,2alpha-dihydroxy-3beta-acetoxyurs-9(11),12-diene (3) on the basis of spectral analyses, including 1D and 2D NMR and mass spectroscopy. It is probable that compounds 2 and 3 are artifacts from dehydration of the corresponding allylic alcohols. 1beta,2alpha,3beta,11alpha Tetrahydroxyurs-12-ene (5), the most abundant compound in the extract, was found to be highly cytotoxic to renal, non-small cell lung, and breast cancer cell lines. PMID- 14738405 TI - Chemokines in liver disease. A new field of study of genetic polymorphism? PMID- 14738406 TI - Role of chemokines in the pathogenesis of liver diseases. AB - OBJECTIVE: while chemokines appear to be important in certain inflammatory disorders, little is known about the role of these proteins in chronic hepatitis C. MATERIAL AND METHODS: firstly, an extensive review of the expression pattern of chemokines in liver diseases with special emphasis in hepatitis C has been performed. Then, expression of selected CXC and CC chemokines and their receptors was assessed by immunohistochemistry and flow cytometry in a number of patients with chronic hepatitis C. RESULTS: evidence that certain CXC and CC chemokines are markedly expressed in chronic hepatitis C has been recently reported. In agreement with this, we have shown that the majority of liver-derived T lymphocytes from patients with chronic hepatitis C expressed CXCR3 and CCR5 chemokine receptors. In addition, an intense intrahepatic expression of their respective ligands, Mig and RANTES, was detected in the same patients studied. Furthermore, an association between the intrahepatic chemokine expression level and the inflammatory activity of chronic hepatitis C was found. CONCLUSIONS: all of these findings strongly suggest that intra hepatic chemokine signaling could play a key role regulating significant pathological events during chronic hepatitis C, opening new avenues for therapeutic interventions based on chemokines activities. PMID- 14738407 TI - Healing of Helicobacter pylori gastric ulcers: only eradication matters. AB - INTRODUCTION: some authors suggest that Helicobacter pylori eradication favors gastric ulcer healing. OBJECTIVE: to study which factors influence ulcer healing in patients suffering from gastric ulcer with H. pylori infection. SUBJECTS AND METHODS: a prospective study of 230 patients with gastric ulcer associated to H. pylori infection. Chronic ingestion of non-steroidal anti-inflammatory drugs was considered as an exclusion. In an initial endoscopy, malignancy was histologically excluded and two biopsies each of antrum and body were obtained. Also, ELISA IgG serology and a 13C-urea breath test were performed. Eradication therapy with omeprazole (20 mg twice a day), clarithromycin (500 mg twice a day) and amoxicillin (1 g twice a day) was administered for seven days, followed by omeprazole 20 mg once a day for five more weeks. Endoscopy was repeated after 6 weeks of treatment and breath test was repeated 2 month after completing therapy. RESULTS: overall gastric ulcer healing was achieved in 80.8% (95% CI: 75-85%) of cases by intention-to-treat, and in 82.6% (77-87%) per protocol. Ulcer healing was achieved in 94.3% (90-97%) of patients with eradication success, but only in 40.8% (28-54%) of patients with eradication failure (p<0.0001). In the multivariate analysis, H. pylori eradication was the only variable that correlated with ulcer healing (odds ratio 24; 95% CI: 10-56; p<0.0001) (x2 model: 64.4; p<0.0001). Additional variables (age, sex, sporadic ingestion of NSAIDs, smoking, previous ulcer disease, ulcer size and location) were not related to healing. CONCLUSION: H. pylori eradication favors ulcer healing in patients with gastric ulcer, which is an argument in favor of the etiological role of the microorganism in this disease. Other factors did not influence ulcer healing. PMID- 14738408 TI - Clinical, anorectal manometry and surface electromyography in the study of patients with fecal incontinence. AB - OBJECTIVE: to demonstrate the role of the clinical, anorectal manometry and surface electromyography in the assessment of patients with fecal incontinence. PATIENTS AND METHODS: ninety-three patients with fecal incontinence are retrospectively reviewed and the data obtained from the directed clinical history, physical examination of the anal region, digital rectal examination, anorectal manometry and surface electromyography are analyzed. A treatment was administered in accordance with the alterations encountered and the results evaluated at 3 and 12 months. RESULTS: fecal incontinence was predominant (91.4%) in women age 59.7+/-11. A background of obstetric risks (48.2%) was frequent in women. Also, 73.1% of the patients presented diarrhea. The anorectal manometry (ARM) demonstrated some alterations in 90.3% of the patients, whereas a hypotonic sphincter was the most common finding (85.7%). Rectal sensitivity or distensibility alterations were present in the rest of the patients. In 79.2% ofthe cases, hypotonic sphincter was associated with rectal sensitivity or distensibility alterations. In 65.2% of patients with hypotonic external anal sphincter, damage of the pudendal nerve was found and therefore biofeedback was indicated in 41.9% of them. CONCLUSIONS: the clinical study of the patients, together with the anorectal manometry and surface electromyography enables the identification of the cause of FI and its treatment. These studies demonstrate that in most cases the origin of the incontinence is due to multiple etiologies, however the treatment of some of the factors involved frequently improves the symptomatology. PMID- 14738409 TI - Approach to a colorectal polyp. PMID- 14738410 TI - Intestinal T-cell lymphoma associated with celiac disease masked by cavernous lymphangioma. AB - We present the case of a patient admitted to our emergency ward with a clinical setting of acute abdominal pain and a history of cavernous lymphangioma, diagnosed in another center by exploratory lapartomy. The patient presented complete analysis including serology tests, as well as an abdominal CT scan that revealed multiple large size retroperitoneal cysts. In view of the clinical symptomatology and results of the tests, a second CT scan was carried out upon admission. As a result of the findings obtained, a second exploratory laparotomy was carried out in which intestinal resection of the perforated jejunal loop and largest cysts was performed. Pathological anatomy diagnosed an intestinal lymphoma associated with enteropathy and abdominal cysts compatible with cavernous lymphangioma. In this work we describe both pathologies, the most characteristic aspects are analyzed and the etiology and possible relation between both entities is discussed. PMID- 14738411 TI - Meckel's diverticulum adenocarcinoma: a rare malignant degeneration. AB - The case of a 40 year old male that, after the etiological study performed due to rectorrhages that was the only symptom he presented, was referred to an exploratory laparotomy. A disseminated adenocarcinoma arising in Meckels diverticulum was detected and palliative segmentary resection of the small bowel was performed. The lack of previous symptoms lead to the dissemination of the disease and to the impossibility of curative surgical resection of the tumor. PMID- 14738412 TI - Liver adenomatosis. PMID- 14738413 TI - [Atypical endoscopic appearance in ulcerative colitis]. PMID- 14738414 TI - Inspiratory flow limitation in obstructive sleep apnoea patients. AB - Patients suffering from the obstructive sleep apnoea syndrome (OSAS) experience nocturnal episodes of upper airway obstruction resulting in recurrent oxygen desaturations and arousals. Methods to quantify the nocturnal obstructive events are of interest for characterizing this prevalent sleep disorder. In a study published in this issue of Clinical Science, Bloch and co-workers propose the computation of a new index for objectively quantifying the degree of flow limitation in patients with OSAS. The results obtained in a bench test and in a pilot study in patients suggest that the flow limitation index proposed may help to better characterize the disturbed breathing events undergone by patients with OSAS. PMID- 14738416 TI - Stopping antiepileptic drugs after successful surgery: what do we know? And what do we still need to learn? PMID- 14738417 TI - Sulthiame in the primary therapy of West syndrome: a randomized double-blind placebo-controlled add-on trial on baseline pyridoxine medication. AB - PURPOSE: West syndrome (WS) is still one of the most difficult to treat epilepsies in infancy. Sulthiame (STM), which is commonly used in some countries in the treatment of benign focal epilepsies in childhood, has been suggested to be effective in WS too. This prospective, randomized placebo-controlled study was designed to prove or refute this hypothesis. METHODS: Thirty-seven infants aged between 3.5 and 15 months with newly diagnosed WS received baseline therapy with pyridoxine (PDX). The children were randomized in a double-blind fashion to STM (n = 20) or placebo (n = 17), starting at day 4 at a moderate dose of 5 mg/kg body weight. Without complete cessation of infantile spasms (ISs) and resolution of hypsarrhythmia as the definition criteria of a response, the dose was doubled at day 7. The final examination was undertaken at the end of day 9. RESULTS: Based on the intention to treat, six (30%) of 20 patients responded to STM (p < 0.025), as did six (35%) of 17 infants fulfilling the study criteria (p < 0.01). Patients with tuberous sclerosis did not respond to STM (n = 3) No patient responded to placebo. One patient in the verum group was withdrawn because of reversible somnolence. CONCLUSIONS: As no child in the placebo group responded to the baseline PDX therapy, nor did any child in the STM group during the first 3 days of baseline therapy, PDX does not seem to be effective in WS. STM has a positive effect in the primary therapy of WS, comparable to that of vigabatrin. PMID- 14738419 TI - Evaluation with alpha-[11C]methyl-L-tryptophan positron emission tomography for reoperation after failed epilepsy surgery. AB - PURPOSE: Reoperation after failed cortical resection can alleviate seizures in patients with intractable neocortical epilepsy, provided that previously nonresected epileptic regions are accurately defined and removed. Most imaging modalities have limited value in identifying such regions after a previous surgery. Positron emission tomography (PET) using alpha-[11C]methyl-L-tryptophan (AMT) can detect epileptogenic cortical areas as regions with increased tracer uptake. This study analyzed whether increased cortical AMT uptake can detect nonresected epileptic foci in patients with previously failed neocortical resection. METHODS: Thirty-three young patients (age 3-26 years; mean age, 10.8 years) with intractable epilepsy of neocortical origin, and a previously failed cortical resection performed at various epilepsy centers, underwent further presurgical evaluation for reoperation. AMT-PET scans were performed 6 days to 7 years after the first surgery. Focal cortical areas with increased AMT uptake were objectively identified and correlated to ictal EEG data as well as clinical variables (age, postsurgical time, etiology). RESULTS: Cortical increases of AMT uptake were detected on the side of the previous resections in 12 cases. In two patients scanned shortly (within a week) after surgery, diffuse hemispheric increases were observed, without any further localization value. In contrast, in 10 (43%) of 23 patients scanned >2 months but within 2.3 years after surgery, focal cortical increases occurred, concordant with seizure onset on ictal EEG. Age, etiology (lesional vs. cryptogenic), epileptiform EEG activity during PET, or time of the last seizure were not significantly related to the presence of increased AMT uptake. All patients with localizing AMT-PET, who underwent reoperation, became seizure free (n = 5) or showed considerable improvement of seizure frequency (n = 2). CONCLUSIONS: AMT-PET can identify nonresected epileptic cortex in patients with a previously failed neocortical epilepsy surgery and, with proper timing for the scan, can assist in planning reoperation. PMID- 14738418 TI - Stimulus-induced rhythmic, periodic, or ictal discharges (SIRPIDs): a common EEG phenomenon in the critically ill. AB - PURPOSE: To describe an underrecognized EEG phenomenon in critically ill patients undergoing continuous EEG monitoring (cEEG). We named these EEG patterns stimulus induced rhythmic, periodic, or ictal discharges (SIRPIDs). METHODS: We reviewed 150 consecutive patients undergoing cEEG during a 9-month period and compared those with and without SIRPIDs. SIRPIDs were defined as periodic, rhythmic, or ictal-appearing discharges that were consistently induced by alerting stimuli. RESULTS: We identified 33 patients with SIRPIDs (22%). SIRPID patterns included periodic epileptiform discharges in 21 patients (nine lateralized) and rhythmic patterns with evolution that fulfilled criteria for ictal discharges in 18 patients (12 unilateral). Eight patients had prior epilepsy; 24 had acute brain injury. Half the patients (17 of 33) had seizures, clinical or subclinical, during the acute illness in addition to SIRPIDs, and half (16 of 33) did not. No significant difference was found in the incidence of clinical seizures in patients with SIRPIDs (30%) compared with those without (45%). Clinical status epilepticus was more common in patients with focal (43%) or ictal-appearing (33%) SIRPIDs than in those without SIRPIDs (17%). CONCLUSIONS: Rhythmic, periodic, or ictal-appearing EEG patterns are commonly elicited by stimulation in critically ill, encephalopathic patients. Recording video, documenting patient stimulation, or repetitively examining patients during cEEG is necessary to recognize these patterns and to differentiate SIRPIDs from spontaneous seizures. Further research is necessary to determine the pathophysiologic, prognostic, and therapeutic significance of SIRPIDs. PMID- 14738420 TI - Resective surgery for intractable focal epilepsy in patients with low IQ: predictors for seizure control and outcome with respect to seizures and neuropsychological and psychosocial functioning. AB - PURPOSE: To investigate possible predictive factors for seizure control in a group of children and adults with low IQs (IQ, < or =70) who underwent resective surgery for intractable focal epilepsy and to study outcome with respect to seizures and neuropsychological functioning. We also studied psychosocial outcome in the adult patients. METHODS: Thirty-one patients (eight children younger than 18 years) with a Wechsler Full Scale IQ of 70 or less underwent comprehensive neuropsychological assessments before and 2 years after surgery. Adults also completed the Washington Psychosocial Seizure Inventory (WPSI). Univariate analyses were used to identify variables differentiating between patients who became seizure free and those who did not. Pre- and postoperative test results were compared by t test for dependent samples. RESULTS: Forty-eight percent of the patients became seizure free, 52% of those with temporal lobe resection and 38% of those with extratemporal resection. Only one variable was predictive for seizure outcome: duration of epilepsy. In one third of the patients, who had the shortest duration of epilepsy (<12 years), 80% became seizure free. Significant improvement was seen regarding vocational adjustment in adults (WPSI). Seizure free adults improved their Full Scale IQ scores. No cognitive changes were found in seizure-free children or in patients who did not become seizure free. CONCLUSIONS: A good seizure outcome was obtained after resective surgery in patients with intractable focal epilepsy and low IQ, provided that treatment was done relatively shortly after onset of epilepsy. No adverse effects were seen on cognitive and psychosocial functioning. PMID- 14738421 TI - Mutations of neuronal voltage-gated Na+ channel alpha 1 subunit gene SCN1A in core severe myoclonic epilepsy in infancy (SMEI) and in borderline SMEI (SMEB). AB - PURPOSE: Severe myoclonic epilepsy in infancy (SMEI) is a distinct epilepsy syndrome. Patients with borderline SMEI (SMEB) are a subgroup with clinical features similar to those of core SMEI but are not necessarily consistent with the accepted diagnostic criteria for core SMEI. The aim of this study was to delineate the genetic correlation between core SMEI and SMEB and to estimate the frequency of mutations in both phenotypes. METHODS: We examined 96 healthy volunteers and 58 unrelated individuals whose clinical features were consistent with either core SMEI (n = 31) or SMEB (n = 27). We screened for genetic abnormalities within exons and their flanking introns of the genes encoding major subunits of the Na+ channels (SCN1A, SCN2A, SCN1B, and SCN2B) by using a direct sequencing method. RESULTS: In both core SMEI and SMEB, various mutations of SCN1A including nonsense and missense mutations were identified, whereas no mutations of SCN2A, SCN1B, and SCN2B were found within the regions examined. All mutations were heterozygous and not found in 192 control chromosomes. Mutations were identified in 26 (44.8%) of the 58 individuals and were more frequent (p < 0.05) in core SMEI (19 of 31) than in SMEB (seven of 27), as assessed by the continuity-adjusted chi2 test. Mutations resulting in a molecular truncation were found only in core SMEI. Among the mutations, two missense mutations were found in both core SMEI and SMEB. CONCLUSIONS: Our findings confirm that SMEB is part of the SMEI spectrum and may expand the recognition of SMEI and suggest other responsible or modifying genes. PMID- 14738423 TI - Status epilepticus in a population-based Virginia twin sample. AB - PURPOSE: To characterize status epilepticus (SE) and estimate its frequency of first occurrence, as well as to assess the contribution of genetic factors to risk of SE occurrence in a sample of Virginia-born twins ascertained from the population-based Mid-Atlantic Twin Registry. METHODS: The occurrence of SE was determined in 13,506 unselected Virginia-born twin pairs ascertained from birth records. Twins included in the study were between ages 2 and 75 years when surveyed. History of seizures and SE was validated through medical records and by detailed personal or parental interviews. RESULTS: Among 381 twins included in 332 pairs with a verified history of seizures, 70 (18.4%) were validated to have had at least one episode of SE. The frequency of first SE in this sample was 309 per 100,000 twins. First SE occurred in conjunction with 21 of 158 febrile and 49 of 223 afebrile seizure cases, respectively. Mean length of SE episode was 76.2 +/- 14.9 min. Age at first SE occurrence ranged from 2 months to 59 years. All concordant twin pairs in the sample were monozygotic (MZ), with a proband-wise concordance rate estimated for SE in this population of 0.31 [95% confidence interval (CI), 0.14-0.52) overall, and 0.67 (95% CI, 0.35-0.90) in pairs concordant for epilepsy. CONCLUSIONS: These results provide a direct estimate of the frequency of SE in a defined population of twins and afford further evidence for a genetic contribution to risk for SE. PMID- 14738422 TI - Generalized epilepsy with febrile seizures plus (GEFS+): clinical spectrum in seven Italian families unrelated to SCN1A, SCN1B, and GABRG2 gene mutations. AB - PURPOSE: We describe seven Italian families with generalized epilepsy with febrile seizures plus (GEFS+), in which mutations of SCN1A, SCN1B, and GABRG2 genes were excluded and compare their clinical spectrum with that of previously reported GEFS+ with known mutations. METHODS: We performed a clinical study of seven families (167 individuals). The molecular study included analysis of polymerase chain reaction (PCR) fragments of SCN1A and SCN1B exons by denaturing high-performance liquid chromatography (DHPLC) and direct sequencing of GABRG2 in all families. We excluded SCN1A, SCN1B, and GABRG2 genes with linkage analysis in a large pedigree and directly sequenced SCN2A in a family with neonatal-infantile seizures onset. We compared the epilepsy phenotypes observed in our families with those of GEFS+ families harboring mutations of SCN1A, SCN1B, and GABRG2 and estimated the percentage of mutations of these genes among GEFS+ cases by reviewing all published studies. RESULTS: Inheritance was autosomal dominant with 69% penetrance. Forty-one individuals had epilepsy: 29 had a phenotype consistent with GEFS+; seven had idiopathic generalized epilepsy (IGE); in three, the epilepsy type could not be classified; and two were considered phenocopies. Clinical phenotypes included FS+ (29.2%), FS (29.2%), IGE (18.2%), FS+ with focal seizures (13%) or absence seizures (2.6%), and FS with absence seizures (2.6%). Molecular study of SCN1A, SCN2A, SCN1B, and GABRG2 did not reveal any mutation. Results of our study and literature review indicate that mutations of SCN1A, SCN2A, SCN1B, and GABRG2 in patients with GEFS+ are rare. CONCLUSIONS: The most frequently observed phenotypes matched those reported in families with mutations of the SCN1A, SCN1B, and GABRG2 genes. IGE and GEFS+ may overlap in some families, suggesting a shared genetic mechanism. The observation that 13% of affected individuals had focal epilepsy confirms previously reported rates and should prompt a reformulation of the "GEFS+" concept to include focal epileptogenesis. PMID- 14738424 TI - Recurrence after a first unprovoked cryptogenic/idiopathic seizure in children: a prospective study from Sao Paulo, Brazil. AB - PURPOSE: To evaluate the recurrence risk after a first unprovoked seizure in a large population of children and adolescents of a developing country. METHODS: This prospective study was conducted at two tertiary hospitals, between September 1989 and August 1998. Children were enrolled if they had a first unprovoked cryptogenic/idiopathic seizure and maximal interval to the enrollment < or =90 days. EEG and computed tomography (CT) were performed in most patients. Potential predictors of recurrence were compared by using the Cox proportional hazards model in univariate and multivariate analyses. Survival analysis was performed by using the Kaplan-Meier curves. RESULTS: Two hundred thirteen children were included. Recurrence occurred in 34% of the patients, and mean time for recurrence was 12 months. Statistical analysis showed significance for seizure recurrence only for patients with abnormal EEGs. CT was performed in 182 patients, and abnormalities were found in 9.5%. Small calcifications were the most frequent finding, and this was not a predictor for recurrence. CONCLUSIONS: The risk of recurrence after a first unprovoked seizure in children from a developing country is similar to that found in developed countries. An abnormal EEG is a risk factor for seizure recurrence in children with a cryptogenic/idiopathic seizure. Calcifications on CT do not increase the risk of recurrence. PMID- 14738425 TI - Direct cost of medical management of epilepsy among adults in Italy: a prospective cost-of-illness study (EPICOS). AB - PURPOSE: To investigate the costs of epilepsy from a nationwide survey comparing adult patients included in different prognostic categories. METHODS: A 12-month prospective observational study was conducted in 15 epilepsy centers from Northern, Central, and Southern Italy. The study population included a random sample of individuals aged 18 years and older with newly diagnosed (ND) epilepsy, seizure remission (R), occasional seizures (OS), active non-drug-resistant (NDR) seizures, drug-resistant (DR) seizures, or surgical candidates (SC). Estimates of the direct costs of care of epilepsy were based on the use of diagnostic examinations, laboratory tests, specialist consultations, hospital admissions, day-hospital days, and drugs, taking the Italian National Health Service perspective. RESULTS: The sample included 631 patients (ND 62, R 158, OS 155, NDR 114, DR 128, and SC 14). The SC group had the highest total cost per patient (3,619 euros) followed by DR (2,190 euros), ND (976 euros), NDR (894 euros), OS (830 euros), and R (561 euros). For each epilepsy group, the main components of the total cost were drugs and hospital admissions. Drug costs increased from the R group to the DR group. The new antiepileptic drugs (AEDs) were the largest part of the cost of treatment. CONCLUSIONS: The costs of epilepsy in referral patients vary significantly according to the time course of the disease and the response to treatment. Hospital admissions and drugs are the major sources of expenditure. PMID- 14738426 TI - Seizure recurrence after planned discontinuation of antiepileptic drugs in seizure-free patients after epilepsy surgery: a review of current clinical experience. AB - PURPOSE: Although epilepsy surgery, especially temporal lobe epilepsy surgery, is well established to control seizures in patients remaining on antiepileptic drug (AED) treatment, less information is available about how many seizure-free surgical patients will relapse after discontinuation of AEDs under medical supervision. METHODS: A literature review yielded six retrospective clinical observations. RESULTS: After planned discontinuation of AEDs in patients rendered seizure free after epilepsy surgery, most often various forms of temporal lobe surgery, the mean percentage recurrence rate in adults in four studies was 33.8%[95% confidence interval (CI), 32.4-35.2%], with maximum follow-up ranging from 1 to 5 years. Seizure recurrence increased during the follow-up of 1 to 3 years and occurred within 3 years of AED discontinuation. In one study of children with temporal lobe epilepsy, the recurrence rate was 20%. More than 90% of adult patients with seizure recurrence regained seizure control with reinstitution of previous AED therapy. Seizure recurrence was unaffected by the duration of postoperative AED treatment; as a consequence, delaying discontinuation beyond 1 to 2 years of complete postoperative seizure control seems to have no added benefit. The occurrence of rare seizures or auras after surgery did not eliminate the possibility of eventual successful AED discontinuation. CONCLUSIONS: AED discontinuation is associated with a seizure recurrence in one in three patients rendered seizure free by epilepsy surgery. These results will be useful in counseling patients about discontinuing AED treatment after successful epilepsy surgery. PMID- 14738427 TI - Factor XIII deficiency associated with valproate treatment. AB - PURPOSE: We present two children who developed a deficiency of factor XIII with valproate (VPA) treatment. This coagulation disorder has not been described in association with VPA treatment in children, and only very recently in one adult patient. RESULTS: Both patients showed recurrent epistaxis as major clinical sign of a combination of decreased coagulation parameters (factor XIII deficiency with thrombocytopenia and decreased von Willebrand factor, respectively). A few days after reduction or withdrawal of VPA treatment, clinical symptoms disappeared, and laboratory findings were within normal range. CONCLUSIONS: VPA is known to influence the synthetic function of the liver and the number and function of megakaryocytes. Therefore an alteration of the factor XIII level by VPA is conceivable. Our case reports suggest that bleeding symptoms during VPA treatment may be caused or aggravated by a decreased factor XIII activity. A determination of factor XIII activity should be considered before surgical procedures during VPA treatment to minimize the risk of (severe) postsurgical bleeding complications. PMID- 14738428 TI - A new benign adult familial myoclonic epilepsy (BAFME) pedigree suggesting linkage to chromosome 2p11.1-q12.2. AB - Benign adult familial myoclonic epilepsy (BAFME) is an autosomal dominant condition characterized by cortical tremor and generalized seizures, mapped on chromosome 8q24 by Japanese authors. Recently the same phenotype also was reported in European families, with linkage on chromosome 2. We present a new family with suggestion of linkage to chromosome 2p11.1-2q12.2 (lod score value, 1.55). This observation would confirm that BAFME is a worldwide, genetically heterogeneous condition, probably with Japanese families linked to 8q24 and European families to 2p11.1-q12.2. PMID- 14738431 TI - Diagnostic performance of the variant lymphocyte flag of the Abbott Cell-Dyn 4000 haematology analyser. AB - BACKGROUND: In addition to differential cell counts, modern haematology analysers generate suspect flags if abnormal cells are detected. Reports on validation of suspect flags are scarce. We have routine experience with the Abbott Cell-Dyn 4000 analyser for over 5 years and have previously demonstrated the utility of the blast flag. Here we report a similar study on the performance of the analyser's Variant Lymphocyte (VL) flag. AIM OF THE STUDY: Evaluation of the diagnostic performance of the Cell-Dyn 4000 VL flag, as compared with lymphocyte morphology in blood smears. In addition, we investigated the usefulness of the numerical VL flag confidence index as provided by the analyser. MATERIALS AND METHODS: All samples generating a VL flag were reviewed over a 5-month period. We also reviewed smears from patients with known lymphoid disorders, even if the analyser did not flag the sample. Two experienced investigators assessed lymphocyte morphology independently. RESULTS: In total, 187 samples were included in the study, of which 183 had a VL flag and four had not. Of the 183 flagged samples, 83 appeared to have abnormal lymphocyte morphology and 100, normal lymphocyte morphology. The sensitivity of the VL flag for detecting abnormal lymphocytes was 0.95 and the positive predictive value was 0.44. Using ROC analysis of the VL flag confidence index, the area under the ROC curve was 0.58 (95% confidence interval 0.50-0.65). CONCLUSIONS: The Cell-Dyn VL flag has reasonable sensitivity but a high false-positive rate. In addition, its performance is insufficient for detecting clinically relevant abnormal lymphocytes. As the VL flag appeared to rely mainly on numerical criteria, it has no added value over numerical criteria defined by the laboratory. PMID- 14738430 TI - Evaluation of the coulter LH 750 haematology analyzer compared with flow cytometry as the reference method for WBC, platelet and nucleated RBC count. AB - The Coulter LH 750 is a new haematology analyser with several new features: a count of nucleated red blood cells (NRBCs), automated WBC correction in presence of a flag indicating a cellular interference and a lower incidence of platelet or WBC interference flags when compared with the GEN.S, our current instrument. We had three main goals in our study: evaluating the LH 750 WBC counts when a GEN.S flag suggests a risk of WBC interference, ascertaining whether the platelet counts not flagged by the LH 750 were accurately assessed in samples flagged by the GEN.S and evaluating the NRBC assay provided by the LH 750. Flow cytometry, using CD45 and CD41, respectively for WBC and platelet labelling, was used as a reference method to assess the accuracy of the LH 750 counts. NRBC were identified by double labelling with propidium iodide (PI) and CD45, NRBCs being CD45-/PI+. A significant relationship was found between LH 750 and flow cytometric WBC counts, whether a WBC correction was made by the LH 750 (r = 0.9809, n = 54) or not (r = 0.9901, n = 23). A highly significant relationship was observed for platelets not only in the range from 0 to 450 x 10(9)/l (r = 0.981, n = 108) but also in cases of thrombocytopenia (range: 0-80 x 10(9)/l; r = 0.956, n = 51). In samples with NRBCs, the NRBC percentages given by the LH 750 and by flow cytometry were highly correlated (r = 0.977, n = 60) and WBC counts were accurate. In conclusion, the reduction in flagging by the LH 750, the accuracy of the results, and the availability of a NRBC count, constitute major advantages. PMID- 14738432 TI - Newborn screening for sickle cell disease in Brazil: the Campinas experience. AB - Newborn screening for sickle cell disease commenced in 1992 in Sao Paulo State and by the end of 2000, the programme covered 78 institutions in 36 municipalities with the screening of 281,884 babies. Initially based on liquid cord blood samples, these are being replaced by dried filter paper capillary samples to ease handling and avoid diagnostic confusion from maternal contamination. The prevalence of sickle cell trait (2.0%) and HbC trait (0.6%) increased significantly between 1996 and 2000, apparently because of improved detection rather than the later introduction of institutions serving populations with higher trait frequencies. There were 29 babies with homozygous sickle cell SS disease and 26 with sickle cell-haemoglobin C (SC) disease, the latter significantly exceeding expectation and possibly attributable to a nonrandom selection of partners. Sickle cell-beta thalassaemia syndromes were proportionately more common than in Jamaica, and it is possible that this results from interaction with other Brazilian populations carrying higher beta thalassaemia gene frequencies. The frequency of abnormal haemoglobins in this population is lower than in Jamaica, but clinically significant sickle cell disease occurred once in every 5527 births, comparable with the frequencies of other significant inborn errors of metabolism. PMID- 14738433 TI - Automated computer result reporting for haemoglobinopathy screening. AB - The anticipated introduction of universal antenatal screening can be expected to increase the workload of haemoglobinopathy laboratories throughout the country. We have devised a rule-based system to process those results that does not require skilled interpretation, thereby freeing staff time for more specialized work. The system relies on a calculated test to create a total haemoglobin peak value, which incorporates the values for HbA, HbA2 and HbF, the MCV and MCH from the full blood count. Each parameter has a series of defined ranges which, when subjected to an interpretation process within the laboratory computer system, generates an automated result text for the sample. During a 6-month verification period, the automated result interpretation system in conjunction with laboratory information systems (LIS) validation reduced the number of samples requiring manual review by 60%. The system was found to be 100% sensitive and 61.8% specific. We feel that the current specificity is acceptable in order to maintain a safe system. The ability to concentrate on potentially abnormal results will allow laboratories and health care workers more time to develop appropriate and timely frameworks to deal with abnormal results. PMID- 14738434 TI - Near-triploidy and near-tetraploidy in hematological malignancies and mutation of the p53 gene. AB - Hyperdiploidy of > or =58 chromosomes is reported in 0.5-3% of hematological malignancies, but reports of near-triploidy (58-80 chromosomes) and near tetraploidy (81-103 chromosomes), are few. We examined these chromosome abnormalities and analyzed the relationship with the mutation of the p53 gene. Thirty-one of 979 adult patients (3.2%) with hematological malignancies were identified as having near-triploid or near-tetraploid (tri-/tetraploid) chromosomes. These included 11 with B-cell neoplasms, seven with Hodgkin's lymphoma, five with T-cell neoplasms, four with myelodysplastic syndromes and four with acute myeloid leukemias. All patients had concurrent complex chromosome aberrations. Deletion of one allele of the p53 gene was found in two patients and a point mutation of the p53 gene was detected in five patients. Although abnormalities of the p53 gene have been reported in about 10% of hematological malignancies, these were found in seven of 31 (23%) patients with tri /tetraploidy. These findings suggest that the abnormality of the p53 gene may be closely related with tri-/tetraploidy. The four myelodysplastic syndrome (MDS) patients with tri-/tetraploidy had a significantly worse prognosis than those with diploid cytogenetics (n = 35; P < 0.002). In B-cell neoplasms (n = 3), triploidy was associated with a worse prognosis than tetraploidy (n = 8) and diploidy (n = 130; P < 0.02). PMID- 14738435 TI - The use of fluorescent molecular beacons in real time PCR of IgH gene rearrangements for quantitative evaluation of multiple myeloma. AB - BACKGROUND AND OBJECTIVES Fluorescent molecular beacons have been employed as hybridization probes in real time quantitative PCR to quantify residual disease in multiple myeloma (MM). DESIGN AND METHODS: After clinical diagnosis of MM, the CDR1, CDR2 and CDR3 regions of the IgH gene were analysed and sequenced to identify its clonal nature. Unique sequences of the clonal IgH rearrangement were used to design specific molecular beacon probes for each MM patient. A molecular beacon probe for the beta-globin gene was used as a reference control to calculate relative amounts of the clonal B-cell population. RESULTS: Optimization of probe design resulted in the use of a competitive sequence at the IgH area target between the loop and part of the stem of the molecular beacon. Cycling conditions and fluorescence temperature acquisition were optimized for a Light Cycler. To validate this method for the follow-up of treated MM patients, we investigated accuracy, as well as interassay and intrassay reproducibility. CONCLUSIONS: Our results indicated that real time PCR with specific molecular beacons provides a feasible, accurate and reproducible method for the determination of minimal residual disease in MM. PMID- 14738436 TI - Diagnosis of plasma cell leukaemia: findings of the UK NEQAS for Leucocyte Immunophenotyping scheme. AB - The diagnosis of plasma cell leukaemia, a rare disorder with an aggressive clinical course and poor prognosis, is not always straightforward and may be dependent on the results of immunophenotyping. Samples from two cases of plasma cell leukaemia have been issued by the UK NEQAS for Leucocyte Immunophenotyping Scheme during the last 5 years and on each occasion a significant number of laboratories failed to make the correct diagnosis. The details of the two samples issued and the results of both surveys are presented. The data highlights the need to adhere to guidelines for immunophenotyping, with respect to using the correct antibody panels, the importance of data interpretation in conjunction with morphological appearance as well as the need to participate in external quality assurance schemes. PMID- 14738438 TI - Accuracy, reproducibility and clinical utility of the CoaguChek S portable international normalized ratio monitor in an outpatient anticoagulation clinic. AB - The accuracy and reproducibility of the CoaguChek S, and its clinical agreement with conventional laboratory international normalized ratio (INR) determination, were evaluated in an outpatient anticoagulation clinic setting. Forty-three patients provided 248 paired INR measurements for analysis. The paired results were highly correlated (r = 0.90). The mean coefficient of variation for the CoaguChek S for a random sample of 21 patients with three repeated tests each, was 4%. Clinical applicability was also measured by discrepant INR values, as defined in the literature by expanded and narrow agreement, and by INR values resulting in a different clinical decision by a blinded haematology registrar. Expanded agreement and narrow agreement between the two INR values occurred 90 and 88% of the time, respectively. The stricter criteria set down by the clinician resulted in 73% of paired results producing the same dosage decision. The CoaguChek S displayed good correlation with laboratory determination of INR and compared relatively well with expanded and narrow clinical agreement criteria. PMID- 14738437 TI - Safe introduction of warfarin for thrombotic prophylaxis in atrial fibrillation requiring only a weekly INR. AB - A new protocol for initiating warfarin therapy was introduced to reduce the workload in the Anticoagulant Clinic. A total of 200 outpatients, with a median age of 74 years, requiring anticoagulation for atrial fibrillation, commenced warfarin 3 mg daily for 1 week. Patients were initially seen weekly, and subsequent warfarin doses were dictated by the International Normalized Ratio (INR) on days 8 and 15; 84% of patients followed the protocol correctly: of these 86% had an INR > or =2 by day 15 and >98% had INR >2 by day 22. By day 22, 58% of patients achieved a stable dose, 85% by day 29 and >95% by day 36. Day 8 INR was predictive of the final maintenance dose required. No patient suffered any thrombotic or haemorrhagic complications in the first month: only three patients had an INR >3 on day 8, and 11 patients had an INR >4 on day 15. Patient age and sex were not sufficiently related to warfarin requirement to provide useful predictive information. This protocol, requiring only weekly INRs, has proved safe and effective for outpatient warfarinization, and has reduced clinic attendances in this population. PMID- 14738439 TI - Validation of a quantitative SPR assay for recombinant FVIII. AB - Surface plasmon resonance was employed to establish a quantitative assay for recombinant FVIII (rFVIII) products using rFVIII as standard. The anti-FVIII monoclonal antibody ESH4 was immobilized onto a carboxymethyldextran surface. A range of rFVIII concentrations were injected over the surface and the binding response enhanced by the addition of a further monoclonal antibody ESH8. Validation using National Institute of Biological Standards and Controls (NIBSC) sixth International rFVIII concentrate standard gave inter- and intra-assay coefficient of variations (CVs) of 7.5 and 3.68% respectively for ESH4-rFVIII binding alone. Enhancement of the binding signal by secondary addition of ESH8 produced inter- and intra-assay CVs of 2.75 and 1.5%. The ESH4 immobilized chip was found to retain binding capacity following regeneration for at least 75 cycles. The assay was found to be unsuitable for quantitation of plasma derived FVIII product but may prove useful for monitoring of rFVIII production. PMID- 14738440 TI - Excellent response of refractory life-threatening thrombotic thrombocytopenic purpura to cyclosporine treatment. AB - The introduction of plasma exchange has significantly improved the outcome of thrombotic thrombocytopenic purpura (TTP), and survival has increased from 10 to 80-90%. TTP refractory to plasma exchange therapy, however, is still a therapeutic challenge. We describe here a patient who partially responded to plasma exchange therapy, but remained totally dependent on plasma infusions. Several attempts to discontinue plasma therapy repeatedly lead to relapses. TTP did not response to vincristine, either. After 3 months treatment with plasma therapy, cyclosporine was started. Plasma therapy could be discontinued after 3 weeks on cyclosporine, and serum LDH and blood platelet count were gradually normalized during 2 months. Cyclosporine was tapered off after 6 months treatment, and the patient has stayed in remission ever since. We conclude that cyclosporine is a worthwhile treatment option in patients with refractory TTP. PMID- 14738441 TI - Protein 4.1 deficiency and deletion of chromosome 20q are associated with acquired elliptocytosis in myelodysplastic syndrome. AB - We report a case of myelodysplastic syndrome (MDS), associated with prominent elliptocytosis. A 66-year-old male presented with peripheral pancytopenia, and was diagnosed with MDS [refractory anaemia (RA)]. Apart from marked elliptocytosis, dyshaematopoietic features were not evident in his peripheral blood or hypercellular bone marrow. After 18 months, he had progressed to RA with excess blasts in transformation. Analysis of red blood cell membrane proteins by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) showed a reduced quantity of protein 4.1 (30% of control). Deletion of chromosome 20q was identified by conventional cytogenetic analysis and fluorescence in situ hybridization. Marked elliptocytosis, persistent for more than 17 months, decreased strikingly after chemotherapy with idarubicin and Ara-C. These findings suggest that acquired elliptocytosis occurred as an unusual morphological feature of MDS, associated with abnormalities of protein 4.1 and chromosome 20q. PMID- 14738442 TI - Aleukaemic leukaemia cutis: case report and review of the literature. AB - 'Aleukaemic leukaemia cutis' or acute leukaemia confined to the skin is extremely rare, although skin involvement with underlying leukaemia is well recognized, and is associated with a poor prognosis. We report a case of isolated acute myeloid leukaemia (AML) in the skin. A literature review shows this to be commonly misdiagnosed. Its recognition is important, because early diagnosis should lead to more appropriate chemotherapy, and a better prognosis. These patients probably require therapy directed specifically to the skin, as well as to other extramedullary sites, such as the central nervous system, to prevent early relapse. PMID- 14738443 TI - Phagocytosis of opsonized apoptotic cells: roles for 'old-fashioned' receptors for antibody and complement. AB - Efficient phagocytic clearance of apoptotic cells is crucial in many biological processes. A bewildering array of phagocyte receptors have been implicated in apoptotic cell clearance, but there is little convincing evidence that they act directly as apoptotic cell receptors. Alternatively, apoptotic cells may become opsonized, whereby naturally occurring soluble factors (opsonins) bind to the cell surface and initiate phagocytosis. Evidence is accumulating that antibodies and complement proteins opsonize apoptotic cells, leading to phagocytosis mediated by well-defined 'old-fashioned' receptors for immunoglobulin-Fc and complement. In this review we summarize the evidence that opsonization is necessary for high capacity clearance of apoptotic cells, which would render putative direct apoptotic cell receptors redundant. PMID- 14738444 TI - Fluvastatin treatment inhibits leucocyte adhesion and extravasation in models of complement-mediated acute inflammation. AB - Complement activation plays a relevant role in the development of tissue damage under inflammatory conditions, and clinical and experimental observations emphasize its contribution to inflammatory vasculitides. Statins have recently been shown to reduce cardiovascular morbidity independently of plasma cholesterol lowering and in vitro studies support a direct anti-inflammatory action of these drugs. The aim of this study was to verify the in vivo effect of fluvastatin on complement-mediated acute peritoneal inflammation. The effect of oral treatment with fluvastatin was investigated in normo-cholesterolaemic rats that received intraperitoneal injection of either yeast-activated rat serum (Y-act RS) or lipopolysaccharide to induce peritoneal inflammation monitored by the number of PMN recruited in peritoneal fluid washes. In addition, vascular adherence and extravasation of leucocytes were evaluated by direct videomicroscopy examination on mesentery postcapillary venules topically exposed to Y-act RS. The number of PMN in the peritoneal washes of rats treated with fluvastatin was 38% lower than that of untreated animals (P < 0.05) 12 h after LPS injection, and was even lower (56%) in rats treated with Y-act RS already 8 h after injection (P < 0.02). Firm adhesion to endothelium and extravasation of leucocytes evaluated under direct videomicroscopy observation were significantly inhibited in fluvastatin treated rats (77% and 72%, respectively; P < 0.01), 120 min after treatment with Y-act RS. Our results demonstrate that fluvastatin inhibits in vivo complement dependent acute peritoneal inflammation and suggest a role for statins in preventing the inflammatory flares usually associated with complement activation in chronic diseases, such as SLE or rheumatoid arthritis. PMID- 14738445 TI - Desferrioxamine modulates chemically induced T helper 2-mediated autoimmunity in the rat. AB - A rise in interleukin (IL) 4-dependent immunoglobulin E (IgE) is a hallmark of the mercuric chloride (HgCl2)-induced Th2-mediated autoimmune syndrome in the Brown Norway (BN) rat, and one of the mediators in allergic asthma in human. Oxidative stress, a potential factor related to the pathogenesis of allergy and asthma, has been shown to up-regulate IL-4 in mast cells and predispose to degranulation in vitro. However, it remains unknown whether oxidative/antioxidative imbalance plays a role in this Th2-driven model of autoimmunity in the rat. Here we show that administration of the non-sulphydryl containing antioxidant desferrioxamine i.p. and s.c. to BN rats reduces HgCl2 enhanced IL-4 gene expression and inhibits HgCl2-induced Th2-mediated autoimmunity. Desferrioxamine treatment suppresses significantly IgE production and lymphoproliferation, and reduces tissue injury in the form of caecal vasculitis in the HgCl2-induced autoimmune syndrome. These results support a role for oxidative stress in the pathogenesis of the HgCl2-induced Th2-dominated autoimmune syndrome. This finding might have implications for understanding the mechanisms involved in Th2 cell responses as seen in allergy and asthma and thereby aid the development of new therapeutic strategies for these diseases. PMID- 14738446 TI - Generation of B220low B cells and production of autoantibodies in mice with experimental amyloidosis: association of primordial T cells with this phenomenon. AB - To investigate the immunological state in amyloidosis, mice were twice intraperitoneally injected (2-week interval) with casein emulsified in complete Freund's adjuvant. Two weeks after the treatment, amyloid deposits were detected in the spleen and other organs of these mice. The number of lymphocytes yielded by the liver and spleen increased significantly. The most affected lymphocyte subset was found to be B cells, namely, the total number of B cells increased and unusual B220low B cells were newly generated in the liver and spleen. In other words, not only normal B220high B cells but also unusual B220low B cells were detected in these organs of mice with amyloidosis. In parallel with this phenomenon, autoantibodies against denatured DNA were detected in sera. Since such autoantibodies are known to accompany the functional activation of NKT cells, NKT cell-deficient mice were used for the induction of amyloidosis. Such mice showed less formation of amyloidosis and lower levels of autoantibodies in sera. Athymic nude mice were NKT cell-deficient but NK1.1- TCRint cells were present. These athymic mice showed an intermediate induction of amyloidosis. The cytokine profile seen in mice with amyloidosis was the Th0 type, showing simultaneous production of IL-4 and IFNgamma. These results suggest that the generation of B220low B cells and the production of autoantibodies in aid of primordial T cells may be major immunological mechanisms in amyloidosis mice. PMID- 14738447 TI - Qualitative and quantitative abnormalities in splenic dendritic cell populations in NOD mice. AB - The phenotype and function of splenic DC populations from diabetes-prone NOD mice were characterized and compared to DC from diabetes-resistant strains in the presence or absence of Flt3 ligand (FL) treatment. NOD mice were found to have significantly fewer CD8alpha+ DC than both B10.BR and C57BL/6 mice, and this defect was reversed by FL treatment. Freshly isolated CD8alpha+ and CD8alpha- DC from all three strains were found to express similar levels of costimulatory molecules and this was similar in both FL-treated and untreated animals. IL-12 p40 production was significantly lower in purified CD11c+ DC from NOD mice compared to DC from C57BL/6 or B10.BR mice. CD8alpha+ DC isolated from NOD mice produced lower levels of IL-12p40 than CD8alpha+ DC from C57CBL/6 and this was dependent on the nature of the stimulus given. In contrast both CD8alpha+ and CD8alpha- DC from FL-treated mice produced high levels of IL-12p40 following activation, but only the CD8alpha- DC produced IL-12p70. Functionally, freshly isolated CD8alpha- DC were more stimulatory than CD8alpha+ DC in a primary allogeneic mixed lymphocyte reaction. However, DC maturation resulted in increased T cell stimulatory capacity for both DC subsets, and this pattern was seen in all strains. These results demonstrate significant differences in phenotype and function of splenic NOD CD8alpha+ DC, and further suggest that FL treatment may reverse some of these abnormalities. PMID- 14738448 TI - Dietary soy phytoestrogens and ERalpha signalling modulate interferon gamma production in response to bacterial infection. AB - Diets rich in soy phytoestrogens have many potential health benefits but isoflavones such as genistein may suppress cell mediated immune function. The effect of dietary phytoestrogens on the host response to infection has not been extensively examined. Mice were fed a diet containing soy phytoestrogens and infected with Mycobacterium avium to establish a chronic infection and inflammatory response. As phytoestrogens may act through classical oestrogen receptors (ER), mice deficient in ERalpha signalling and wild type mice were evaluated for a panel of Type 1-associated cytokines (IFNgamma, IL-12 and IL-18) in the spleen. IFNgamma production in the spleen was increased approximately 4 fold in ERalpha-deficient mice fed a casein-based diet over wild type mice fed a casein-based diet (P < 0.05), suggesting a role for ERalpha in suppressing IFNgamma production. IL-18 levels in spleens of wild type mice were decreased compared to ERalpha-deficient mice on a casein diet. Splenic IL-12 and IL-18 levels were not affected in wild type and ERalpha-deficient mice on the phytoestrogen containing diets, with the exception that whole soy increased IL-12 levels in the tissues of ERalpha deficient mice. We conclude that ERalpha and dietary phytoestrogens can influence production of key regulatory cytokines in response to chronic bacterial infection. PMID- 14738449 TI - Endotoxin stimulates monocyte-endothelial cell interactions in mouse intestinal Peyer's patches and villus mucosa. AB - Although monocyte-endothelial cell interactions represent an initial step in controlling the recruitment of monocytes in inflamed tissues, their dynamic processes in microvessels of lymphoid (Peyer's patches) and non-lymphoid (villus) regions in gut-associated lymphoid tissue remain poorly understood. We monitored the migration of fluorescence-labelled monocytes derived from the spleen in intestinal microvessels with or without lipopolysaccharide (LPS) treatment and investigated the role of adhesion molecules, P-selectin, vascular cell adhesion molecule (VCAM-1) and intercellular adhesion molecule-1 (ICAM-1). In control mice, there were few interactions between infused monocytes and the endothelium of intestinal microvessels. The monocyte-endothelial interactions (both rolling and adhesion) were significantly increased in intestinal microvessels of LPS treated mice compared with those in controls. Anti-P-selectin monoclonal antibody (MoAb) significantly suppressed the LPS-induced increase in monocyte rolling in postcapillary venules of Peyer's patches and submucosal venules. Anti-VCAM-1 MoAbs significantly suppressed the LPS-induced increase in monocyte adhesion to postcapillary venules (PCVs) of Peyer's patches, submucosal venules, and villus capillaries. In contrast, anti-ICAM-1 MoAb significantly suppressed the number of adherent monocytes in PCV of Peyer's patches but not in submucosal venules or villus capillaries. These observations demonstrated that LPS treatment resulted in a significant increase in recruitment of monocytes both in microvessels of lymphoid and non-lymphoid regions and that P-selectin, VCAM-1 and ICAM-1 appeared to play important roles in LPS-induced interactions. PMID- 14738450 TI - High dose interleukin-12 exacerbates Bordetella pertussis infection and is associated with suppression of cell-mediated immunity in a murine aerosol challenge model. AB - The in-vivo clearance of Bordetella pertussis infections in murine models in naive mice and animals vaccinated with whole-cell vaccine is considered to be via a Th-1-dependent mechanism in which interleukin-12 (IL)-12 may play a prominent role. It has also been demonstrated clearly that the treatment of animals with macrophage-derived IL-12 administered with an acellular vaccine can increase the efficacy of this vaccine preparation to levels seen with the whole-cell vaccine. However, the effects of exogenously added IL-12 on immune responses in non vaccinated B. pertussis-challenged mice remain unclear, with two studies giving contradictory findings. In this study we have treated mice with escalating doses of mIL-12 (0.1-10 microg/mouse) prior to challenge with B. pertussis (using an aerosol challenge model of infection). The ability of mice to clear infection was assessed in IL-12 treated and in phosphate buffered saline (PBS) control animals at days 6 and 13 post-challenge. Lymphoid cells were isolated from spleen and cell-mediated immune responses assessed at days 1, 6 and 13 post-challenge. In addition, the direct effects of high-dose IL-12 on challenged mice was assessed by checking natural killer (NK) activity from isolated lung and spleen lymphoid cells as well as interferon-gamma (IFN-gamma) generation from isolated cells and serum at day 1 post-challenge. The results from this study show that bacterial colonization of the lungs is actually enhanced following treatment with high-dose IL-12. This is associated with impaired cellular immune responses. The mechanisms associated with the immunosuppressive effects of IL-12 are discussed. PMID- 14738451 TI - Stimulation of innate immunity by susceptible and multidrug-resistant Pseudomonas aeruginosa: an in vitro and in vivo study. AB - In attempt to investigate the stimulatory effect of Pseudomonas aeruginosa on innate immunity and to correlate it to its level of resistance to antimicrobials, 20 isolates were applied; 8 isolates were susceptible and 12 multidrug-resistant. Genetic diversity was defined by PFGE. Human monocytes of two healthy volunteers were in vitro stimulated by the isolates for the production of pro-inflammatory (TNF-alpha, IL-1beta, IL-6, IL-8 and IL-12) and anti-inflammatory cytokines (IL 10), of malondialdehyde and of procalcitonin. Cytokines were estimated by EIA, malondialdehyde by the thiobarbiturate assay and procalcitonin by an immunochemiluminometric assay. Survival of 48 Wistar rats was recorded after induction of sepsis by the intraperitoneal injection of three susceptible and three multidrug-resistant isolates. To test whether comparative effect of the latter isolates on survival correlates with any difference of monocyte-mediated release of pro-inflammatory mediators, monocytes of two rats were in vitro stimulated for the production of TNF-alpha and of malondialdehyde. In vitro stimulation of human monocytes by the susceptible isolates elicited elevated production of malondiadeheyde, of IL-1beta and of IL-6 compared to stimulation by multidrug-resistant isolates. Similar differences were found for TNF-alpha and IL 8, but they were not statistically significant. Production of IL-10 and IL-12 was not detected after stimulation with any isolate. Levels of procalcitonin were similar after induction with either susceptible or multidrug-resistant isolates. Mean survival of animals was 7.56, 21.80 and 55.20 h, respectively, after challenge by the susceptible isolates and 28.89, 61.8 and more than 120 h, respectively, after challenge by the multidrug-resistant isolates. Differences of survival were accompanied by greater rodent monocyte-release of TNF-alpha and malondialdehyde after stimulation by the susceptible isolates compared to multidrug-resistant ones. It is concluded that considerable differences are encountered on the stimulation of human monocytes by susceptible and resistant isolates of Pseudomonas aeruginosa. These results correlate with in vivo evidence and might influence decision on therapeutics. PMID- 14738452 TI - Detection of HLA class II-dependent T helper antigen using antigen phage display. AB - Major histocompatibility complex (MHC) class II-dependent antigens not only activate CD4+ T helper (Th) cells, but also cytolytic T lymphocytes and effector cells of the innate immune system. These antigens therefore are candidate vaccines against cancer and infectious agents. We have developed a novel approach using a model antigen, tetanus toxoid (TT), which provides the basis for the establishment of a novel strategy of cloning Th antigens. In the TT model system, a cDNA library encoding part of the TT light chain which contained a TT associated Th epitope recognized by TT-specific Th clones was displayed on a phage vector (TT-phage) and presented to TT-specific Th cells by autologous Epstein-Barr virus-transformed B cells (APC). These TT-phages were able to specifically activate two different TT-specific CD4+ Th cell lines as demonstrated both in [3H]thymidine incorporation and cytokine release assays. Th cell stimulation by TT-phages was significant at a ratio of one TT-phage in 50 irrelevant phages. The described approach provides the basis for the development of a novel strategy of cloning MHC class II-dependent Th antigens, using available Th cells. This strategy has several potential advantages over existing antigen cloning methods or biochemical peptide isolation. PMID- 14738453 TI - Blocking B7 and CD40 co-stimulatory molecules decreases antiviral T cell activity. AB - Inhibition of co-stimulatory signals for T cells by interrupting CD80/CD86-CD28 and CD40-CD154 interactions is a promising approach to prevent transplant rejection and to induce graft tolerance. However, this tolerizing treatment might affect T cell reactivity towards all the antigens to which the immune system is exposed during treatment. We addressed the question whether such inhibition of co stimulatory ligands on human antigen presenting cells (APC) would affect T cell reactivity against a virus. This was tested in an in vitro system with freshly isolated human monocytes transduced with adenovirus (ad) containing either murine interferon-gamma (mIFN-gamma) or green fluorescent protein (GFP) as marker transgene. T cells co-cultured with transduced monocytes proliferated and produced cytokines. These 'primed' T cells had strong antiviral activity as they subsequently killed ad/GFP-transduced monocytes and reduced mIFN-gamma accumulation in coculture with ad/mIFN-transduced monocytes. However, if priming had occurred in the presence of blocking anti-CD40/CD80/CD86 MoAbs, generation of this antiviral activity was completely prevented. Moreover, T cells primed in the absence of co-stimulatory cells failed to proliferate upon restimulation with adenovirus-transduced monocytes. The results confirm that co-stimulatory signals from APC are required for efficient induction of antiviral T cell activity and point to a potential infectious risk of blocking co-stimulatory signals. PMID- 14738454 TI - Expression of CTLA-4 in nonhuman primate lymphocytes and its use as a potential target for specific immunotoxin-mediated apoptosis: results of in vitro studies. AB - T-cell-mediated immunoregulation is one of the main mechanisms implicated in induction and maintenance of transplantation tolerance. In this regard, deletion or modulation of xeno/alloantigen-specific T cells, as well as blocking of their interactions with other cell populations, are currently being pursued for tolerance induction in humans as well as nonhuman primates. In order to investigate whether cytotoxic T-lymphocyte antigen-4 (CTLA-4) may represent a suitable target for a T cell depletion approach in nonhuman primate models, we analysed CTLA-4 expression in peripheral blood mononuclear cells (PBMCs) from nonhuman primates and the potential role of two anti-CTLA-4 saporin-conjugated immunotoxins. The analysis was performed in PBMCs from 8 cynomolgus monkeys from Philippines and from Mauritius both at protein level by flow cytometry and at transcriptional level by RT-PCR. In addition, the apoptotic role of the immunotoxins was investigated. The results showed that CTLA-4 was expressed at variable levels depending on the origin of the cynomolgus monkeys and the resting or activated cell condition. CTLA-4 was not expressed on resting Mauritius PBMCs and showed a lower up-regulation upon PMA/PHA activation compared to the Philippines PBMCs that expressed CTLA-4 also before activation. Two CTLA-4 RNA transcripts (672 and 550 bp) were detected with levels variations after cell stimulation. Two anti-CTLA-4 immunotoxins induced in vitro apoptosis of activated PBMCs from both sources of cynomolgus monkeys. This is the first report that documents CTLA-4 expression both at protein and transcriptional level by nonhuman primate PBMCs and provides novel perspectives of xeno/allograft rejection immunotherapy based on CTLA-4 targeting. PMID- 14738455 TI - Increased Toll-like receptor 4 expression in infants with respiratory syncytial virus bronchiolitis. AB - The fusion protein of the respiratory syncytial virus (RSV) binds to the pattern recognition receptors, TLR4 and CD14, and initiates innate immunity response to the virus. The aim of the study was to investigate the expression of TLR4 on peripheral blood lymphocytes and monocytes in peripheral blood of infants in both acute and convalescent phase of RSV bronchiolitis (n = 26). In addition, TNF alpha expression in lipopolysaccharide-stimulated monocytes was also assessed. The results showed TLR4 to be expressed predominantly by monocytes in both sick infants and controls. During the acute phase of infection monocytes up-regulated TLR4 in eight infants, which returned to the levels recorded in controls 4-6 weeks from infection. There was no difference in the percentage of TNF-alpha secreting monocytes. Of the clinical parameters tested, minimal oxygen saturation was found to correlate negatively with this expression in the group of infants with increased TLR4. Additional studies are under way to correlate this finding with the outcome of the immune response to RSV. PMID- 14738456 TI - Low plasma level of adiponectin is associated with stavudine treatment and lipodystrophy in HIV-infected patients. AB - This study tested the hypothesis that in patients with HIV-associated lipodystrophy, adiponectin levels were related to insulin resistance, TNF-alpha and IL-6 and treatment with nucleoside analogues. HIV seropositive men undergoing highly active antiretroviral treatment were enrolled into three predetermined clinical groups: lipodystrophy with central fat accumulation (n = 12); lipodystrophy without central fat accumulation (n = 15); no lipodystrophy (n = 15). HIV-negative healthy men served as controls (n = 12). Both lipodystrophic groups had a low percentage of limb fat compared to the two control groups. Patients with lipodystrophy with fat accumulation had increased truncal fat compared with controls. Levels of adiponectin did not correlate with either TNF alpha or IL-6. Low levels of adiponectin were found in both lipodystrophic groups and were associated with current or previous treatment with stavudine. Furthermore, the adiponectin level correlated with the percentage of limb fat. Patients with lipodystrophy with fat accumulation were more insulin resistant, measured by HOMA-IR, compared with controls. However, HOMA-IR did no correlate to adiponectin or other cytokines. In conclusion, the finding of no difference between the two lipodystrophic groups with regard to adiponectin, indicates that low levels of adiponectin reflects fat atrophy, whereas the insulin resistance was best explained by increased truncal fat mass. PMID- 14738457 TI - Interleukin-15 production by monocyte-derived dendritic cells and T cell proliferation in HIV-infected patients with discordant response to highly active antiretroviral therapy. AB - A discordant response to highly active antiretroviral therapy (HAART) occurs when CD4 T cell counts are stable or increased over time despite persistently detectable HIV-RNA levels. In order to identify immunological factors affecting discordant treatment responses, a total of 27 HIV-infected patients were studied: (a) 10 naive patients (mean CD4+ = 101.5 cells/microl; mean HIV-RNA = 4.8 log10 copies/ml); (b) seven responder patients (mean CD4+ = 908.9 cells/microl); and (c) 10 discordant patients (mean CD4+ = 396.1 cells/microl; mean HIV-RNA = 5.4 log10 copies/ml). Five healthy blood donors were included as HIV-seronegative controls. The following parameters were evaluated: interleukin (IL)-15 production by monocyte-derived dendritic cells (MDDC) after stimulation with lypopolysaccaride (LPS) and Candida albicans; recall and HIV-1-specific antigen lymphocyte proliferation (LP). Increased levels of IL-15 production by MDDC after stimulation with LPS and C. albicans were found both in discordant patients and responder patients. Conversely, a strong reduction of IL-15 levels was observed in naive patients. Discordant patients developed positive LP responses to C. albicans and HIV-1 p24. LP in response to C. albicans and HIV-1 p24 was also positive in responder patients. Decreased LP response was found in naive patients. In conclusion, HIV-infected patients with discordant viro-immunological responses to HAART present increased levels of IL-15 production by MDDC and enhanced recall and HIV-1-specific antigen LP responses, suggesting an improvement in indices of immune function. PMID- 14738458 TI - Cellular immunity induced by the recombinant Plasmodium falciparum malaria vaccine, RTS,S/AS02, in semi-immune adults in The Gambia. AB - Vaccination of malaria-naive humans with recombinant RTS,S/AS02, which includes the C-terminus of the circumsporozoite protein (CS), has been shown to induce strong T cell responses to both the whole protein antigen and to peptides from CS. Here we show that strong T cell responses were also observed in a semi-immune population in The Gambia, West Africa. In a Phase I study, 20 adult male volunteers, lifelong residents in a malaria-endemic region, were given three doses of RTS,S/AS02 at 0, 1 and 6 months. Responses to RTS,S, hepatitis B surface antigen and peptides from CS were tested using lymphocyte proliferation, interferon (IFN)-gamma production in microcultures, and IFN-gamma ex vivo and cultured ELISPOT, before and after vaccination. Cytotoxic responses were tested only after vaccination and none were detected. Before vaccination, the majority of the volunteers (15/20) had detectable responses in at least one of the tests. After vaccination, responses increased in all assays except cytotoxicity. The increase was most marked for proliferation; all donors responded to RTS,S after the third dose and all except one donor responded to at least one peptide after the second or third dose. There was a lack of close association of peptide responses detected by the different assays, although in microcultures IFN-gamma responses were found only when proliferative responses were high, and responses by cultured ELISPOT and proliferation were found together more frequently after vaccination. We have therefore identified several peptide-specific T cell responses induced by RTS,S/AS02 which provides a mechanism to investigate potentially protective immune responses in the field. PMID- 14738459 TI - High levels of type 2 cytokine-producing cells in chronic fatigue syndrome. AB - The aetiology of chronic fatigue syndrome (CFS) is not known. However, it has been suggested that CFS may be associated with underlying immune activation resulting in a Th2-type response. We measured intracellular production of interferon (IFN)-gamma and interleukin (IL)-2; type 1 cytokines), IL-4 (type 2) and IL-10 (regulatory) by both polyclonally stimulated and non-stimulated CD4 and CD8 lymphocytes from patients with CFS and control subjects by flow cytometry. After polyclonal activation we found evidence of a significant bias towards Th2- and Tc2-type immune responses in CFS compared to controls. In contrast, levels of IFN-gamma, IL-2 and IL-10-producing cells were similar in both study groups. Non stimulated cultures revealed significantly higher levels of T cells producing IFN gamma or IL-4 in CFS patients. Concluding, we show evidence for an effector memory cell bias towards type 2 responsiveness in patients with CFS, as well as ongoing type 0 immune activation in unstimulated cultures of peripheral blood cells. PMID- 14738460 TI - Paneth cell granule depletion in the human small intestine under infective and nutritional stress. AB - Paneth cells are important contributors to the intestinal antimicrobial barrier through synthesis and release of antimicrobial peptides and proteins. Animal studies indicate that Paneth cell numbers, location and granule morphology are altered by infection and zinc status. We examined human tissue to determine whether Paneth cell numbers, distribution or granule morphology are altered in infective, inflammatory and nutritional disorders. Archival sections from infective disorders (giardiasis, cryptosporidiosis, HIV, helminth infection) were compared with active inflammatory conditions (coeliac, Crohn's and graft-versus host diseases) and histologically normal tissues. A subset of tissues was studied by electron microscopy and TUNEL staining for apoptosis. Human defensin-5 (HD5) peptide and mRNA was analysed by immunohistochemistry, in situ hybridization and quantitative reverse transcription polymerase chain reaction. Sections from a tropical population cohort study were then analysed to determine the relationship of granule depletion to infection, nutritional status and plasma zinc concentration. In HIV-related cryptosporidiosis, but not other disorders, Paneth cells were reduced in number and markedly depleted of granules. Paneth cell granule depletion was associated with reduced HD5 immunoreactivity, but this was not due to apoptosis and there was no reduction in mRNA transcripts. In the tropical population studied, depletion of granules was associated with reduced body mass index, reduced plasma zinc levels and HIV infection. Paneth cell granules in human small intestine may be depleted in response to infective and nutritional stress. We postulate that this is one mechanism through which zinc status influences host susceptibility to intestinal infection. PMID- 14738461 TI - Involvement of interleukin 18 in Crohn's disease: evidence from in vitro analysis of human gut inflammatory cells and from experimental colitis models. AB - An imbalance of immunoregulatory factors and/or cells contributes to uncontrolled mucosal T cell activation and inflammation in Crohn's disease (CD). Bioactive interleukin (IL)-18 has been shown to be produced by macrophages in CD lesions. We report here that T cells freshly isolated from inflamed tissue of CD patients (and not T cells from control intestinal tissue) were responsive to IL-18. In the presence of IL-18, these T cells produced more interferon (IFN)-gamma and less IL 10. To analyse further the role of IL-18 in this disease, an acute and a chronic model of murine colitis were used. IL-18 mRNA was significantly enhanced in trinitrobenzene sulphonic acid (TNBS) induced colitis, and treatment with IL-18 binding protein (IL-18BPa), which neutralizes IL-18 bioactivity, significantly reduced the severity of colitis. However, IL-18BPa did not affect the course of chronic colitis in CD45RBhighCD4+ T cell reconstituted SCID mice. Production of IFN-gamma in lamina propria mononuclear cell cultures from IL-18BPa-treated SCID mice was decreased, but at the same time fewer lamina propria CD4+ T cells harvested from IL-18BPa-treated mice compared to non-treated mice were in apoptosis. We conclude that IL-18 clearly has a modulatory role in the inflammatory cascade of CD and experimental colitis by affecting IFN-gamma and IL 10 production, and apoptosis. In view of the divergent effects of IL-18 neutralization in the two different murine colitis models, it is unlikely that IL 18 is at the top of this cascade. PMID- 14738462 TI - Immune responses to glutamic acid decarboxylase and insulin in patients with gestational diabetes. AB - Pregnancy is a natural state of immunoprotection and tolerance. We studied subjects with gestational diabetes (GDM) to evaluate the influence of pregnancy on the humoral immune response to the autoantigen GAD and to injected insulin. Antibodies against glutamic acid decarboxylase (GADA) subclasses and epitope reactivity were determined in 34 GADA-positive pregnant patients with GDM, in 20 GADA-positive relatives of people with TID and in 25 GADA-positive patients with newly diagnosed TID. Partum levels of insulin antibodies (IA), IgG1- and IgG4-IA were measured in 131 women with GDM treated with human insulin from the time of diabetes diagnosis (including 22 with GADA) and were compared to 19 patients with TID after 3 months of insulin treatment. GADA titre and subclasses were similar among all groups. GADA in GDM patients bound fewer epitopes than GADA in relatives of patients with TID (all epitopes being present in 23%versus 65%, P < 0.01). In particular, antibodies to the minor GADA epitopes GAD6596-249, GAD651 100 and GAD67 were less frequent in patients with GDM compared to relatives (P < 0.01). Antibodies to insulin (IA) were found in 17% of patients with GDM. They were more frequent in GDM patients with GADA compared to GADA-negative patients (41%versus 12%, P < 0.005). IgG1 was the dominant insulin antibody subclass response in both patients with GDM and TID but levels of IgG1-IA and IgG4-IA were significantly lower in patients with GDM compared to patients with TID (P < 0.004). Antibody responses in women with gestational diabetes appear to be dampened and restricted, but without change in subclass usage. PMID- 14738463 TI - IL-12 enhances the generation of tumour antigen-specific Th1 CD4 T cells during ex vivo expansion. AB - CD4+ T cells are essential for the immune response against cancer. Vaccination against cancer will likely only be effective at preventing growth of micrometastatic disease while adoptive T cell therapy will be better suited for eradication of bulky pre-existing disease (Knutson et al. Expert Opin Biol Ther 2002; 2:55-66). Problems with the use of adoptive T cell therapy include lack of CD4+ T cell help, low frequency of antigen-specific T cells, and lack of effective ex vivo expansion techniques. In this study, we focused on improving ex vivo expansion of CD4+ T helper cells. The effects of IL-12, along with IL-2, on the ex vivo generation of HER-2/neu antigen-specific T cells were examined. Patients were immunized with a peptide-based vaccine that contained a helper epitope, p776-790, derived from the intracellular domain of HER-2/neu. While T cell immunity to p776-790, assessed by proliferation assays, could be readily measured in short-term cultures, cell line generation by multiple in vitro stimulation with peptide and IL-2 as the only added cytokine resulted in loss of antigen-specific proliferation. The inclusion of IL-12, along with IL-2, restored antigen-specific proliferation in a dose-dependent fashion. The resulting p776 790-specific T cells responded readily to antigen by proliferating and producing type I cytokines (IFN-gamma and TNF-alpha). The increased proliferative response of the cultures was due in part to an increase in the number of HER-2/neu specific T cells. These results suggest that IL-12 is an important cytokine for ex vivo recovery and maintenance of antigen-specific CD4+ T lymphocytes that would otherwise be lost by using IL-2 alone in combination with antigen. Furthermore, these results have important implications for ex vivo expansion of CD4+ T cell for use in anti-tumour adoptive immunotherapy. PMID- 14738464 TI - Toll-like receptor (TLR) 2 and 4 mutations in periodontal disease. AB - Toll-like receptors (TLR) are signal molecules essential for the cellular response to bacterial cell wall components. Different functional effective polymorphisms for the TLR 4 gene (Asp299Gly; Thr399Ile) and for the TLR 2 gene (Arg677Trp, Arg753Gln) have recently been described that are associated with impaired lipopolysaccharide signal transduction. A total of 122 patients with chronic periodontal disease and 122 healthy unrelated controls were genotyped for the Asp299Gly and Thr399Ile polymorphism of the TLR 4 gene and the Arg677Trp and Arg753Gln mutation of the TLR 2 gene. The mutations were identified with polymerase chain reaction followed by restriction fragment length polymorphism (RFLP) analysis. The prevalence of the Asp299Gly and the Thr399Ile mutant allele was 4.1% (10/244) and 4.5% (11/244) among periodontitis patients. For the healthy controls the prevalence was 3.3% (8/244) for the Asp299Gly (P = 0.810) and 3.7% (9/244) for the Thr399Ile mutant allele (P = 0.819). The Arg753Gln mutant allele was found in 2.9% (7/244) of the periodontitis subjects as compared to 4.1% (10/244) in the control group (P = 0.622). The Arg677Trp mutant allele was not found in any of the study subjects. Unlike in ulcerative colitis there was not observed an association between chronic periodontitis and the various mutations of the TLR 2 and 4 gene. PMID- 14738465 TI - Mycobacterial heat shock protein 70 induces interleukin-10 production: immunomodulation of synovial cell cytokine profile and dendritic cell maturation. AB - Cytokines are key modulators of the immune responses that take place in the inflamed synovium of arthritis patients. Consequently, substances that can reverse the inflammatory profile of the inflamed joint are potential tools for clinical management of the disease. Mycobacterial heat shock protein 70 (MTBHSP70) has been found to protect rats from experimentally induced arthritis through the induction of interleukin (IL)-10-producing T cells. In this study, we have demonstrated that MTBHSP70 induces IL-10 production in synoviocytes from arthritis patients and peripheral blood monoculear cells (PBMCs) from both patients and healthy controls. IL-10 production was accompanied by a decrease in tumour necrosis factor (TNF)-alpha production by synovial cells. Separation studies showed that the target cells were mainly monocytes. Accordingly, we observed that MTBHSP70 delayed maturation of murine bone marrow-derived dendritic cells. Our results suggest that MTBHSP may act on antigen-presenting cells (APCs) to modulate the cytokine response in arthritis and support an anti-inflammatory role for this protein, suggesting that it may be of therapeutic use in the modulation of arthritis. PMID- 14738470 TI - The effect of physical activity on endothelial function in man. AB - Endothelial dysfunction occurs early in the atherosclerosis in response to elevated atherosclerotic risk factors, and endothelial dysfunction itself may exacerbate the atherosclerotic process. Treatments that reduce atherosclerotic risk factors also generally improve endothelial function. The present review seeks to summarize the effect of exercise training on endothelial function in human subjects. Cross-sectional studies comparing healthy physically active and inactive subjects as well as longitudinal exercise training studies of healthy individuals show little effect of exercise training on endothelial function. In contrast, both cross-section and longitudinal studies document improved endothelial function with exercise training in subjects with abnormal baseline endothelial function, including the elderly and patients with heart failure or coronary artery disease. Improvements in endothelial function with physical activity may explain some of the benefits of exercise in subjects with, or at risk for, vascular disease. PMID- 14738471 TI - Effect of the cytoskeletal fixation agent phalloidin on transcapillary albumin transport and interstitial fluid pressure following subdermal prostaglandin E1 administration in the rat. AB - AIM: Lowering of interstitial fluid pressure (Pif) facilitates fluid filtration across the capillary membrane and oedema formation in acute inflammation. The cellular mechanism behind this lowering of Pif involves beta1-integrins mediating contact between dermal cells and the extracellular matrix fibres, and also the cell cytoskeleton as disruption of actin filaments using cytochalasin-D induced a lowering of Pif and oedema formation. Fixation of actin with phalloidin attenuates oedema formation and abolishes lowering of Pif in anaphylaxis in the rat. The objective of this study was to determine whether phalloidin modifies lowering of Pif and albumin extravasation in rat skin also after prostaglandin E1 (PGE1). METHODS: Pif was measured using micropipettes connected to a servo controlled counterpressure system. Microvascular permeability was estimated as the albumin extravasation (Ealb) using radiolabelled human serum albumin. RESULTS: Subdermal injection of PGE1 (0.85 mg mL-1) lowered Pif from -0.8 +/- 0.8 mmHg (SD) in control to -3.5 +/- 0.9 mmHg (P < 0.05) within 30 min. Pre-treatment with phalloidin (500 microg kg-1) before PGE1 resulted in Pif of -1.7 +/- 1.0 mmHg (P < 0.05 compared with PGE1). Ealb after subdermal saline was 0.07 +/- 0.04 mL g-1 DW and increased to 0.32 +/- 0.32 mL g-1 DW with PGE1 (P < 0.05) but was unaffected by pre-treatment with phalloidin given before PGE1 0.32 +/- 0.35 mL g 1 DW (P > 0.05 compared with PGE1 alone). CONCLUSION: These results are consistent with the concept that the cytoskeleton actin filaments participate in control of Pif. PMID- 14738472 TI - Leukaemia inhibitory factor alters expression of genes involved in rat cardiomyocyte energy metabolism. AB - AIM: Cardiac remodelling is associated with changes in contractile proteins and their performance, alterations in energy production and intracellular calcium homeostasis, as well as changes in extracellular matrix proteins. Some of these processes may be mediated through the gp130 receptor complex. Patients with heart failure have increased cardiac gene expression of leukaemia inhibitory factor (LIF), a cytokine that signals through the gp130 receptor. The aim of this study was to identify alterations in gene expression in LIF-stimulated neonatal cardiomyocytes. METHODS: Cardiomyocytes were isolated from 1- to 3-day-old Wistar rats and stimulated for 48 h with LIF. Gene expression was examined by repeated cDNA filter array analysis (n = 5) and key results verified by complementary methods. RESULTS: In LIF-stimulated cultures we observed increased cell area and changes in gene expression. The intracellular signal regulators signal transducer and activator of transcription 3, calcium/calmodulin-dependent protein kinase IV, protein kinase Cdelta and the transcription factor ID1 were upregulated. Adenylyl cyclase V was downregulated. LIF also induced altered expression of tissue inhibitor of metalloproteinase-1. Receptor genes for tumour necrosis factor, interleukin-4, neurotensin and somatostatin were upregulated. Finally, LIF reduced the expression of components in the adenosine triphosphate synthase complex, epidermal fatty acid-binding protein and insulin-like growth factor binding proteins 1 and 6. CONCLUSIONS: Array analysis revealed changes in mRNA levels of several genes not previously associated with activation of the gp130/LIF receptor complex. Our findings indicate a role for LIF in regulation of cardiomyocyte energy metabolism. PMID- 14738473 TI - Immune system alteration in response to two consecutive soccer games. AB - AIM: Changes in leucocyte and monocyte subpopulations were investigated in 10 elite male soccer players aged 16-19 years. The purpose was to perform a descriptive study of immunological alterations in elite soccer players in response to two consecutive games separated by 20 h. It was hypothesized that in response to two games the players would show signs of short-term immunosuppression. METHODS: Blood samples were taken before the first soccer game, immediately after the second game and after 6, 24, 48 and 72 h. Cell surface antigens, testosterone and cortisol were investigated. RESULTS: During the first 6 h after the second game there was a significant increase in number of circulating neutrophils, mature (CD20+ CD5+) B cells and CD4/CD8 ratio. A significant decrease was observed in the number of natural killer (NK) cells, monocytes and adhesion on lymphocytes and monocytes. In a delayed phase, 48 h after the second game the expression of both adhesion and signalling molecules increased on lymphocytes and monocytes. Changes in adhesion and signalling molecules at 48 h correlated negatively to the subjects VO2max, suggesting larger immunological response to similar exercise in subjects with lower aerobic exercise capacity. CONCLUSION: In response to competitive soccer exercise some immunological variables are enhanced while others are depressed. Observed changes may serve a purpose in adaptation to exercise by signalling via adhesion. PMID- 14738474 TI - Responses of rat myocardial antioxidant defences and heat shock protein HSP72 induced by 12 and 24-week treadmill training. AB - AIM: The purpose of this study was to determine the effects of both a short (12 weeks) and a long-term (24 weeks) endurance treadmill-training programme on the levels of oxidative stress markers, the activity of the enzymatic antioxidants, and the content of the 72 kDa heat shock protein (HSP72) in rat myocardium. METHODS: Thirty male Wistar rats were randomly assigned to exercise trained (n = 16) and sedentary (n = 14) groups. After 12 week of training, eight rats were killed while the remaining eight continued the training programme until 24 week. RESULTS: Seven sedentary controls were killed together with each trained group. Levels of thiobarbituric acid-reactive substances (TBARS), protein carbonyls, and total and oxidized glutathione (tGSH and GSSG) in myocardial homogenates were unchanged by training irrespective of the protocol duration. However, an increased content of the oxidative stress biomarkers was detected in hearts from both the 24-week trained rats and their sedentary controls when compared with their corresponding 12-week groups. The antioxidant enzymatic activities total and mitochondrial superoxide dismutase (tSOD and mtSOD, respectively), glutathione peroxidase (GPX) and glutathione reductase (GR), remained unchanged after the 12-week training period whereas a significant increase in tSOD and mtSOD activities (18%, P < 0.05) was observed in heart homogenates of 24-week trained animals as compared with their sedentary controls. HSP72 expression levels were not significantly modified after 12 week of training but a threefold increase was detected after 24 week (P < 0.05). CONCLUSION: These results demonstrate that a long-term endurance training (24 weeks) induced discrete increases in antioxidant enzyme activities in rat myocardium and elicited a marked enhancement in HSP72 expression levels. However, a shorter training programme (12 weeks), was not effective in increasing heart antioxidant defences. PMID- 14738475 TI - Inhibition of nitric oxide synthesis potentiates the colonic permeability increase triggered by luminal bile acids. AB - AIM: Experiments were performed in anaesthetized rats to clarify the role of nitric oxide (NO) in the control of colonic permeability. METHODS: Colonic luminal pressure, the transmucosal potential difference (PD) and the clearance of [3H] mannitol and [14C] urea from blood to lumen were measured. NO synthesis was blocked with Nomega-nitro-L-arginine (L-NNA) i.v. and mucosal permeability was increased by deoxycholic acid (DCA, 4 mm). The involvement of histamine in the response was studied by giving the histamine H1 receptor blocker pyrilamine. RESULTS: In proximal colon, L-NNA per se increased luminal pressure and PD but had no significant effect on clearance. DCA per se increased luminal pressure, had no significant effect on PD, but increased mannitol and urea clearance and the clearance ratio. L-NNA and pyrilamine both blocked the luminal pressure effect of DCA but L-NNA had no significant effect on the clearance response to DCA. In distal colon, L-NNA per se had no significant effect on pressure and clearance, but increased PD like in proximal colon. DCA had no significant effect on luminal pressure, but markedly reduced PD and increased both clearance and clearance ratio. In this segment, L-NNA significantly potentiated the clearance response to DCA, and further increased clearance ratio to a value not significantly different from unity (1.00 +/- 0.05). CONCLUSION: The data suggest that in vivo, moderate concentrations of bile acids increase colonic permeability in rats via a mechanism that is inhibited by NO in distal but not in proximal colon. In distal colon, NO may contribute to the maintenance of epithelial barrier function. PMID- 14738476 TI - Evidence for intestinal oxidative stress in obstructive jaundice-induced gut barrier dysfunction in rats. AB - AIM: An important factor that promotes bacterial and endotoxin translocation in obstructive jaundice is intestinal injury that causes increased permeability. However, little is known of the submicroscopic biochemical events leading to defects of the intestinal barrier. This study was undertaken to investigate the effect of experimental obstructive jaundice on intestinal lipid peroxidation, protein oxidation and thiol redox state. METHODS: Rats were randomly divided into controls, sham operated and bile duct ligated (BDL). After 10 days, intestinal barrier function was assessed by measuring endotoxin in portal and aortic blood. Tissue samples from the terminal ileum were examined histologically and morphometrically, while other samples were homogenized for the determination of lipid peroxidation, protein oxidation and thiol redox state [reduced glutathione (GSH), oxidized glutathione (GSSG), total non-protein mixed disulphides (NPSSR), protein thiols (PSH) and protein disulphides (PSSP)]. RESULTS: Obstructive jaundice compromised intestinal barrier function leading to significant portal and systemic endotoxaemia. The intestinal mucosa in jaundiced rats was atrophic with significantly decreased villous density and total mucosal thickness. Determination of biochemical parameters of oxidative stress in the intestine showed increased lipid peroxidation and protein oxidation in BDL-rats. Thiol redox state revealed the presence of intestinal oxidative stress in jaundiced rats, indicated by a decrease in GSH and increased GSSG, NPSSR and PSSP. CONCLUSIONS: This study shows that experimental obstructive jaundice induces intestinal oxidative stress, which may be a key factor contributing to intestinal injury and leading to endotoxin translocation. PMID- 14738477 TI - Changes in PCr/Cr ratio in single characterized muscle fibre fragments after only a few maximal voluntary contractions in humans. AB - AIM: This methodological study investigated the number of brief maximal voluntary isometric contractions (MVC) needed to show evidence of fibre activation, as indicated by changes in the phosphocreatine to creatine (PCr/Cr) ratio. METHODS: Subjects performed series of four, seven and/or 10 MVC (1 s on, 1 s off) of the m. quadriceps (60 degrees -flexion angle). Biopsy samples of the m. vastus lateralis were taken at rest and immediately post-exercise. Single muscle fibres were dissected from the freeze-dried samples and classified as types I, IIA or IIAX, using mATPase stainings. Fragments of characterized fibres were analysed for PCr and Cr content. Analyses of variance were performed to investigate changes in PCr/Cr per fibre group over time, followed by Bonferroni post-hoc test (P < 0.01). The fifth percentile of resting values of each fibre group was determined. RESULTS: Mean PCr/Cr ratio after four, seven and 10 MVCs were significantly lower for all fibre groups (P < 0.01). The mean decreases were 44, 64 and 76%, respectively. However, only after seven and 10 contractions PCr/Cr ratios of all, but three type I and two type IIAX fibres, individual fibres were below the fifth percentile. CONCLUSION: In very short duration exercise, involving seven brief maximal voluntary contractions, changes in the PCr/Cr ratio indicated activation of different characterized muscle fibre fragments. The results suggest that this approach may be useful for investigating the pattern of fibre type activation in exercise of very short duration. PMID- 14738478 TI - Effects of prolonged exercise and recovery on sarcoplasmic reticulum Ca2+ cycling properties in rat muscle homogenates. AB - AIM: To examine the effects of exercise and exercise plus active and passive recovery on sarcoplasmic reticulum (SR) Ca2+-handling properties. METHODS: Crude muscle homogenates were prepared from adult rat gastrocnemius muscle from two experiments. In one experiment, the muscle was extracted immediately after prolonged treadmill running (RUN), after a 45 min period of reduced exercise intensity (RUN+) following RUN and compared with controls (CON). In the second experiment, muscle was extracted during passive recovery following the same run protocol at 10 min (REC10), 25 min (REC25) and 45 min (REC45) and compared with CON. RESULTS: Sarcoplasmic reticulum Ca2+-uptake was 31% higher (P < 0.05) in RUN+ compared with CON and RUN. Higher values (P < 0.05) were also found in REC25 (48%) and REC45 (50%) compared with CON. Maximal Ca2+-ATPase was increased by 23% (P < 0.05) in RUN+ compared with CON and RUN and by 65-68% (P < 0.05) in REC25 and REC45 compared with CON. A higher (P < 0.05) Hill coefficient for Ca2+-ATPase activity was observed in RUN+ (2.3 +/- 0.2) compared with CON (1.7 +/- 0.2) or RUN (1.6 +/- 0.2), but not for any REC conditions. In addition, the coupling ratio (Ca2+-uptake/Ca2+-ATPase activity) was higher (P < 0.05) in RUN+ (2.2 +/- 0.10) compared with CON (1.9 +/- 0.05) and RUN (1.9 +/- 0.08). CONCLUSIONS: It is concluded that in crude homogenates, SR Ca2+-uptake and Ca2+-ATPase activity are elevated in recovery following prolonged running and that the elevation in these properties is more pronounced during passive compared with active recovery. PMID- 14738479 TI - Evidence that the Na+-K+ leak/pump ratio contributes to the difference in endurance between fast- and slow-twitch muscles. AB - AIM: Muscles containing predominantly fast-twitch (type II) fibres [ext. dig. longus (EDL)] show considerably lower contractile endurance than muscles containing mainly slow-twitch (type I) fibres (soleus). To assess whether differences in Na+-K+ fluxes and excitability might contribute to this phenomenon, we compared excitation-induced Na+-K+ leaks, Na+ channels, Na+-K+ pump capacity, force and compound action potentials (M-waves) in rat EDL and soleus muscles. METHODS: Isolated muscles were mounted for isometric contractions in Krebs-Ringer bicarbonate buffer and exposed to direct or indirect continuous or intermittent electrical stimulation. The time-course of force decline and concomitant changes in Na+-K+ exchange and M-waves were recorded. RESULTS: During continuous stimulation at 60-120 Hz, EDL showed around fivefold faster rate of force decline than soleus. This was associated with a faster loss of excitability as estimated from the area and amplitude of the M-waves. The net uptake of Na+ and the release of K+ per action potential were respectively 6.5- and 6.6-fold larger in EDL than in soleus, which may in part be due to the larger content of Na+ channels in EDL. During intermittent stimulation with 1 s 60 Hz pulse trains, EDL showed eightfold faster rate of force decline than soleus. CONCLUSION: The considerably lower contractile endurance of fast-twitch compared with slow-twitch muscles reflects differences in the rate of excitation-induced loss of excitability. This is attributed to the much larger excitation-induced Na+ influx and K+ efflux, leading to a faster rise in [K+]o in fast-twitch muscles. This may only be partly compensated by the concomitant activation of the Na+-K+ pumps, in particular in fibres showing large passive Na+-K+ leaks or reduced content of Na+ K+ pumps. Thus, endurance depends on the leak/pump ratio for Na+ and K+. PMID- 14738480 TI - Regulatory gene expression in skeletal muscle of highly endurance-trained humans. AB - AIM AND BACKGROUND: Changes in regulatory and structural gene expression provide the molecular basis for the adaptation of human skeletal muscle to endurance exercise. HYPOTHESIS: The steady-state levels of multiple mRNAs mainly involved in regulatory functions differ between highly endurance-trained and untrained subjects in a muscle heavily recruited during the exercise. METHODS: Biopsies from musculus vastus lateralis of seven untrained (UT) subjects [maximal oxygen consumption (VO2max) = 39 mL kg-1 min-1] and seven trained (T) professional cyclists (VO2max = 72 mL kg-1 min-1) were analysed for the contents of 597 different mRNAs using commercially available cDNA arrays (Clontech no. 7740-1). Intra-individual expression profiles were compared by least-square linear regression analysis. Differences in gene expression between the two groups were tested for statistical significance using L1 regression analysis combined with the sign test on all permutations of scatter plots of log raw values from UT vs. T subjects. RESULTS: Transcripts for 144 of 597 genes were sufficiently abundant to be analysed quantitatively. The expression profiles of the T group had a better intragroup correlation (R2) than those of the UT group (0.78 vs. 0.65, P < 0.05). An intergroup (T vs. UT) correlation of expression profiles gave an R2 of 0.71. Statistical analysis at a false discovery rate of 5% identified differential expression of nine cell-regulatory genes between T and UT. The mRNA levels of eight genes, including two DNA repair enzymes, transcription factors, signal transducers, a glycolytic enzyme and a factor involved in steroid hormone metabolism were increased in T vs. UT. Conversely, the mRNA of the tumour suppressor APC was downregulated with endurance training. Selective reverse transcriptase polymerase chain reaction experiments confirmed the signal estimates from the array analysis. CONCLUSIONS: The repetitive impact of the complex exercise stimuli in professional cyclists attenuated the interindividual differences in regulatory gene expression in skeletal muscle. Long-term nuclear reprogramming of regulatory gene expression seems to be characteristic of human musculus vastus lateralis in a highly endurance-trained steady state. PMID- 14738482 TI - Histocompatibility and grafting. PMID- 14738483 TI - Features of the development of a welfare system for visually disabled people in Denmark. AB - PURPOSE: To describe the development of a formal system of welfare for blind and partially sighted people in Denmark. METHODS: The principal laws in Denmark relating to a formal system of welfare for blind and partially sighted people are noted and commented on. No such collection of laws has been found in the literature. The history of aid to visually disabled people in Denmark is described, as are the Danish classifications of visual impairment. DEVELOPMENT: Formalized welfare for blind and partially sighted people was started in Denmark in 1811, as a private initiative by the Kjaede Order. Later, the Danish state took over responsibility, and in 1858 the Royal Danish Institute for the Blind was established. Since then a series of laws concerning welfare for blind and partially sighted people have been issued in accordance with economic and social development in Denmark. In 1858 the Braille writing system for blind people was introduced. The Danish Association of the Blind was founded in 1911 and has profoundly influenced subsequent legislation. During the last 10 years, several visual centres have been established. Since 1968 prevention of blindness has also played a prominent role in Denmark. CONCLUSION: Great steps have been taken towards improving the welfare of blind and partially sighted people. However, being blind is still very difficult. Modern technologies and hectic lifestyles have created new problems for blind people. The obligations of the Danish state towards visually disabled people have, therefore, increased in recent years. PMID- 14738484 TI - HLA class I/II matching and chronic endothelial cell loss in penetrating normal risk keratoplasty. AB - OBJECTIVE: Chronic endothelial cell loss of the graft is very common after penetrating keratoplasty. The aetiology of this is unknown. Clinically, non identifiable immune reactions have been suspected. Recently, we were able to demonstrate that proper human leucocyte antigen (HLA) matching is a suitable means to reduce classical immune reactions in normal risk keratoplasty patients. In this study, we therefore investigated whether HLA-matched grafts also experience less chronic endothelial cell loss. METHODS: A homogenous group of 223 normal risk keratoplasty patients was divided into six groups with different degrees of HLA matching (group 1 with unknown HLA data, group 2 with up to two mismatches, group 3 with three mismatches, group 4 with four mismatches, group 5 with five mismatches and group 6 with six mismatches on the HLA A, B, DR loci). All serological HLA A, B, C and all moleculargenetic HLA DRB, DRQB typings of donors and recipients were performed in a single laboratory accredited by the American Society for Histocompatibility and Immunogenetics. Only patients with at least three postoperative endothelial cell density values were included in the study. The slopes of the regression lines for each individual scatterplot of endothelial cell density values plotted against postoperative time (linear regression, lost cells/mm2/day), and after logarithmic transformation (exponential regression, annual relative loss of cells) were evaluated, respectively. RESULTS: There were no statistically significant differences between the six groups. CONCLUSION: Whereas proper HLA matching at present standards is already a suitable means to reduce identifiable immune reactions and to prolong graft survival even in normal risk keratoplasty patients, the same HLA matching procedures are not effective in reducing the extent of chronic endothelial cell loss. For several reasons this does not yet exclude, however, the possibility that the underlying cause of chronic endothelial cell loss is immunological. PMID- 14738485 TI - Visual acuity and refractive errors in a suburban Danish population: Inter99 Eye Study. AB - PURPOSE: The present study was performed as part of an epidemiological study, the Inter99 Eye Study. The aim of the study was to describe refractive errors and visual acuity (VA) in a suburban Danish population. METHODS: The Inter99 Eye Study comprised 970 subjects aged 30-60 years and included a random control group as well as groups at high risk for ischaemic heart disease and diabetes mellitus. The present study presents VAs and refractive data from the control group (n = 502). All subjects completed a detailed questionnaire and underwent a standardized general physical and ophthalmic examination including determination of best corrected VA and subjective refractioning. RESULTS: Visual acuity 0.4) differences between the lesion sizes and DTH responses of the tUCA, vehicle only or sham treatment groups. These results demonstrate that topical exposure to UVR-UCA promotes M. ulcerans infection and suppresses DTH responses to M. uclerans antigens in infected animals. These results lend credence to the hypothesis that UVR-mediated enhancement of Buruli ulcer disease in the Crl:IAF(HA)-hrBR hairless guinea-pig model occurs via modulation of cis-urocanic acid-susceptible immune pathways. PMID- 14738529 TI - Photodynamic therapy with chlorin e(6) for skin metastases of melanoma. AB - BACKGROUND: Photodynamic therapy (PDT) has been successfully applied in clinical settings to destroy neoplasms, but the efficacy of such a treatment is dependent on the type of neoplasm and the photosynthesizer used. Here, we perform a clinical assessment of PDT for skin metastases of pigmented melanoma using chlorin e(6). STUDY DESIGN/MATERIALS AND METHODS: PDT with chlorin e(6) photosensitizer was administered to 14 patients with skin metastases from melanoma (10 females, four males, mean age 49.6 years). Chlorin e(6) at a dose of 5 mg/kg of patient's weight was intravenously injected. The treatment course consisted of two courses of PDT exposure 1 h after intravenous chlorin e(6) injection and 24 h post-injection. The light energy density for each skin tumor was 80-120 J/cm(2) per treatment, with a light power density of 250-300 mW/cm(2). RESULTS: All skin melanoma metastases that received PDT showed complete regression with no recurrence during the study period. The complete response of all skin metastases from melanoma occurred in eight cases after one PDT treatment. In the remaining six individuals, tumors required multiple PDT courses prior to complete regression. No cases of photodermatitis were registered. The Karnofsky performance scale score of the patients with skin metastases from melanoma showed no significant difference before and after PDT. No patients had significant changes in blood cell counts that would indicate chlorin e(6) systemic toxic effect. Blood chemistry and urinalysis did not show any evidence of chlorin e(6) renal and hepatic injury. CONCLUSIONS: PDT with chlorin e(6) for skin metastases from melanoma is effective and well tolerated. Further clinical investigation of PDT with chlorin e(6) is warranted. PMID- 14738530 TI - Effects of repeated sunbed exposures on the human skin. In vivo measurements with confocal microscopy. AB - BACKGROUND: Ultraviolet (UV) lamps used in commercial sunbeds are usually defined as UVA sources. Although it is well accepted that sunbed exposure significantly increases melanin pigmentation, its capacity to induce epidermal thickening is discussed controversially. OBJECTIVES: The aim of this study was to assess non invasively the effects of repeated sunbed exposures on epidermal thickness, cell size, and pigmentation by means of confocal laser-scanning microscopy (CLSM) in vivo. METHODS: Eight volunteers had sunbed exposures six times in a 3-week period (cumulative dose: 126 J/cm(2) UVA). During irradiation, a small site (2 cm x 2 cm) on the lateral aspect of the inner forearm was covered with a UV-opaque sheet (non-exposed site). CLSM was performed with the Vivascope (Lucid, Henrietta, NY, USA) 24 h after the last UVA exposure on non-exposed sites and UVA-exposed sites that were on the medial aspect of the inner forearm at a distance of 2 cm to the non-exposed measurement site. The following parameters were assessed: thickness of the horny layer (DSC), minimal thickness of the epidermis (E(min)), minimal thickness of the viable epidermis (VE(min)), cell size of the granular layer (A(gran)), and the epidermal melanin content (MI). Additionally, colorimetric measurements have been carried out on non-exposed and UVA-exposed sites. RESULTS: DSC of the UVA-exposed skin was significantly higher than the one of non-exposed sites (mean+/-SD: 15+/-2.9 microm vs. 12.8+/-3 microm). Although E(min) was significantly higher in UVA-exposed sites (mean+/-SD: 40.4+/-3.6 microm vs. 39+/ 2.9 microm), a slight but not statistically significant (P>0.05) decrease of VE(min) was observed (25.5+/-2.1 microm vs. 26.2+/-2.4 microm). The median of cell size of the granular layer (A(gran)) significantly (P=0.008) differed between non-exposed (752.1 microm(2)) and UVA-exposed sites (600 microm(2)). MI was significantly (P=0.014) higher for the UVA-exposed skin (1.12 vs. 1.34). Accordingly, colorimetry revealed significantly (P< 0.01) lower skin brightness for UVA-exposed sites (L*=60.2+/-4.3) as compared with non-exposed sites (L*=63.4+/-3.9). CONCLUSIONS: Sunbed exposures seem to induce photoadaptation not only by skin pigmentation but also by epidermal thickening that is predominantly due to an increase in thickness of the horny layer. Moreover, our data indicate that UVA radiation has an influence on the cell size of the granular layer. CLSM is a promising tool for photobiological studies in vivo. PMID- 14738531 TI - Skin autofluorescence as a biological UVR dosimeter. AB - BACKGROUND/PURPOSE: Collagen is one of the major endogenous skin fluorophores. Alteration in the structure of collagen due to chronic ultraviolet radiation (UVR) exposure may influence the intensity of the autofluorescence. The aim of this study was to investigate the relation between collagen-linked autofluorescence and sun exposure to clarify whether the skin can be used as a biological UVR dosimeter. METHODS: We conducted an in vivo study with 131 healthy volunteers. Fluorescence was measured from sun-exposed (dorsal forearm, forehead and shoulder) and sun-protected (buttock) skin and corrected for the impact of pigmentation and redness. The excitation wavelengths (Ex) and emission wavelengths (Em) were: Ex330:Em370, Ex330:Em455 and Ex370:Em455 nm. Individual UVR exposure data were collected both retrospectively and prospectively using questionnaires and electronic personal UVR dosimeters for a summer period. RESULTS: Age, but not sex, skin type or smoking habits correlated significantly positively with skin autofluorescence at Ex370:Em455 at all body sites (P<0.001, r(2)=0.08-0.26), and at Ex330:Em455 only at the buttock (P=0.001, r(2)=0.08), whereas age was not correlated with Ex330:Em370. Sun-protected buttock skin had significantly higher autofluorescence than sun-exposed skin (P-values<0.0001). Because of great between-subject differences in autofluorescence at different body sites, and because the autofluorescence at the unexposed buttock represents the baseline value, individual correction of skin autofluorescence measurement with that of the buttock was performed. Different measures of individual chronic cumulative UVR doses correlated significantly negatively with the skin autofluorescence ratio (F(ratio)), but the correlations were poor (r(2)=0.03 0.10). CONCLUSION: The results indicate that the collagen-linked skin F(ratio) might be best to use as a measure of individual photodamage, a UVR dose effect, and that it is also a better marker of individual cumulative UVR dose than the used UVR exposure measurements. The methods used to obtain UVR exposure data might not be sensitive and specific enough. PMID- 14738532 TI - Ocular lens blue autofluorescence cannot be used as a measure of individual cumulative UVR exposure. AB - BACKGROUND/PURPOSE: The accumulation of fluorophores in the ocular lens with age might be caused by ultraviolet solar radiation (UVR) exposure, but evidence of a relation between individual cumulative UVR exposure and lens autofluorescence is lacking. Individually determined UVR exposure has never before been related to lens autofluorescence, and the aim of this study was to investigate if ocular lens blue autofluorescence can be used as a biological UVR dosimeter. METHODS: Ocular lens autofluorescence was quantified in vivo by fluorescence spectroscopy in 145 volunteers (108 healthy subjects, 18 with basal cell carcinoma (BCC) and 19 with cutaneous malignant melanoma (MM)). The excitation wavelength was 350 nm and the fluorescence emission was 450 nm. Individual UVR exposure data were collected both retrospectively and prospectively using questionnaires and electronic personal UVR dosimeters. RESULTS: Lens blue autofluorescence increased significantly with age (P=0.01), and females had significantly higher autofluorescence than males (P=0.024); the two factors explained 10% of the total variation in lens autofluorescence. Neither smoking habits nor use of glasses/contact lenses or sunglasses influenced autofluorescence. No correlations between autofluorescence and UVR exposure measurements were found, and neither was there a difference in autofluorescence between groups with high and low UVR exposure (P-values>0.1), respectively. MM patients had significantly (P=0.019) higher autofluorescence than healthy subjects when age and sex differences were taken into account; no such difference (P=0.097) was detected between BCC patients and healthy subjects. CONCLUSION: The results indicate that age and gender only play a minor role in the level of lens blue autofluorescence. Exposure to UVR has been suggested to be responsible for a part of the age related increase in autofluorescence, but this could not be confirmed in this study. The higher level of lens autofluorescence found in MM patients might be due to genetics rather than higher cumulative UVR exposure. In conclusion, ocular lens blue autofluorescence cannot be used as a biological UVR dosimeter. PMID- 14738533 TI - Iron chelation can modulate UVA-induced lipid peroxidation and ferritin expression in human reconstructed epidermis. AB - BACKGROUND/PURPOSE: As ferritin has been identified as an important factor in antioxidant defense in cultured human skin cells, we evaluated UVA-induced lipid hydroperoxides (LPO) production and ferritin expression in reconstructed human epidermis in vitro. RESULTS: Ferritin is regularly present in the basal layer of unirradiated epidermis both in the human skin in vivo and in the reconstructed human epidermis in vitro. Following acute UVA exposure, ferritin expression increased in basal epidermal cells in both models. Quantitative analysis showed that, in reconstructed human epidermis, LPO and ferritin levels increased linearly with the UVA dose. An iron chelator, OR10141, inhibited these inductions. CONCLUSION: These findings demonstrate that reconstructed human epidermis is a useful in vitro model to study UVA-induced oxidative stress and protection afforded by iron chelators, antioxidants or UVA absorbers. PMID- 14738534 TI - Effect of clothing varieties on solar photosynthesis of previtamin D3: an in vitro study. AB - BACKGROUND/PURPOSE: Vitamin D3 plays important roles in the absorption of calcium and phosphorus from the gastrointestinal tract and in the treatment of rickets; in addition, it facilitates the deposition of minerals in bones, thus minimizing the possibility of developing osteomalacia. Sunlight naturally induces vitamin D3 photosynthesis. Such a process is affected by a number of factors such as age, geographical location, skin color, sunscreen application and clothing. It is intended in the present investigation to study in vitro the effect of clothing on the solar photoproduction of vitamin D3. METHODS: Fifteen different fabric samples were tested for their effect on the efficiency of the in vitro solar conversion of 7-dehydrocholesterol (7-DHC) to vitamin D3. 7-DHC was dissolved in methanol to give a concentration of 2.6 x 10(-4) M. Solutions were exposed to sunlight in quartz containers for predetermined periods either uncovered or covered with the fabric sample under test. Changes in the concentrations of 7-DHC and the photoproducts were monitored by HPLC. Fabrics were graded as the number of threads per square inch (in(2)), and their sunlight attenuation was determined. RESULTS: 7-DHC is transformed to previtamin D3 upon exposure to sunlight, and the amount generated exhibited an almost linear relationship. When fabric-covered samples of 7-DHC were irradiated, photoproducts were also detected and their concentrations depended on the degree of sunlight attenuation imposed by the fabric. Generally, the higher the number of threads per in(2) the more the light attenuation produced. CONCLUSION: Clothing plays an important role in attenuating sunlight, thus leading to diminished vitamin D3 production to an extent that would require dietary compensation. PMID- 14738535 TI - Erythrodermic chronic actinic dermatitis responding only to topical tacrolimus. AB - Chronic actinic dermatitis (CAD) is a disorder characterized by an often severe persistent eczematous eruption induced by exposure to ultraviolet radiation. Treatment involves photoprotective measures and topical corticosteroid therapy and in more severe cases, systemic immunosuppression. Occasionally, however, the condition can prove very resistant to all therapy and be severely disabling. We report a patient with CAD who resisted standard topical and systemic treatments, but responded to topical tacrolimus ointment 0.1% (Protopic). PMID- 14738536 TI - Light-induced seborrhoeic eczema: severe photoprovocation from subclinical disease. PMID- 14738537 TI - Narrow band UVB phototherapy and oral acyclovir for pityriasis rosea. PMID- 14738538 TI - Chronic actinic dermatitis developing during narrowband UVB phototherapy for psoriasis. PMID- 14738540 TI - Genetic epidemiology: some special contributions of birth cohorts. AB - Birth cohorts that collect detailed phenotypic and environmental information, particularly if multigenerational, have an important contribution to make as we seek to understand the genetic and environmental determinants of common disease. Provided such multigenerational population cohorts are unselected by disease, trait or exposure, they can also contribute important information that is needed for optimal statistical analysis in genetic epidemiology generally. One example of this added value is an assessment, at specified loci, of whether or not there is distortion of the expected Mendelian 50 : 50 transmission of alleles to study subjects (e.g. due to differential loss of embryos of one genotype). Another example is to test for heterosis, i.e. whether heterozygotes have a greater or lesser effect than either homozygote. Finally, phenome scans (a fixed format analysis of the associations between a genotype of interest and thousands of outcome variables from the cohort database) could be used as a screening tool to test whether certain classes of genetic variation have more impact than others on human health and development, and so inform genotyping strategies generally. PMID- 14738541 TI - Fever - time for a reprise? PMID- 14738542 TI - Preventing mother-to-child HIV transmission in the new millennium: the challenge of breast feeding. AB - Short courses of antiretroviral drugs have greatly enhanced the prospect of reducing mother-to-child HIV transmission. Yet transmission by breast feeding clouds hopes for this seemingly simple intervention. We revisit mathematical models to assess the competing risks associated with feeding by breast vs. formula. These indicate that, in the less developed world where the HIV epidemic predominates, neither option, unmodified, offers a reasonable choice for HIV positive women. Where infant mortality rates are greater than about 40 per 1000 live births, if formula were made available to HIV-infected women only, the excess number of deaths that would result from formula use would be approximately the same or greater than the number of HIV infections that might be prevented. Only at lower infant mortality rates, less than about 40 per 1000, is the risk greater on the breast. There are thus no good grounds for the total avoidance of breast feeding under all conditions. Research to develop and test safer infant feeding alternatives is an urgent priority. On the one hand, ways to reduce HIV transmission while preserving breast feeding, as exclusive breast feeding could do, need to be fully tested. On the other hand, ways to reduce non-HIV morbidity and mortality associated with formula feeding, as educational or sanitary interventions could do, equally need testing. With either approach, a necessary foundation for implementing all the core components of preventing mother-to-child HIV transmission is competent counselling for mothers. Innovative approaches are needed to mobilise and train effective counsellors among health care workers and, as appropriate, community members. PMID- 14738543 TI - Effect of indoor environmental factors on development of atopic symptoms in children followed up to 4 years of age. AB - The increased prevalence of atopic diseases reported in many Western countries is thought to be caused by changes in living conditions. For a long time attention was focused on indoor environmental factors and early feeding patterns, but during recent years other factors have received more attention. An unselected, Swedish, population-based cohort of 904 children were followed from the age of 3 months to 4 years. Questionnaires were sent to the mothers of the children when they were 3 months, 18 months and 4 years of age. A blood sample was taken from a subgroup of the children when they were 4 years old which was analysed for IgE specific antibodies to food and inhalant allergens. The prevalences of asthma, allergic eye-nose reactions, eczema, and food reactions at 4 years of age and a positive blood test were analysed as outcome factors. Features of housing and early feeding patterns were found to have limited effect on the development of allergies in the children. Presence of older siblings resulted in a decreased tendency to produce IgE antibodies. Early exposure to furry animals seemed to prevent the development of asthma. Children who were frequently infected up to 18 months of age showed an increased prevalence of allergic symptoms at 4 years of age. Children of mothers with a higher prevalence of complaints about indoor factors, or who had increased general symptoms or skin or mucous membrane symptoms ran a greater risk of developing various allergic symptoms. However, these children did not have higher levels of IgE antibodies. The reported allergic symptoms in the children may be due to non-immunological reactions, although it is also possible that the mothers may have over-reported such symptoms. PMID- 14738544 TI - Association of head circumference at birth among sibling pairs. AB - The objective of the study was to estimate and compare the correlation coefficients of head circumference and weight at birth among sibling pairs. Pairs of singleton siblings were ascertained among children born in Norway to the same mother between 1978 and 1997. Head circumference, birthweight and other perinatal factors were registered in the Medical Birth Registry of Norway. Head circumference measurements were obtained for first- and second-born in 287 448 sibling pairs. The correlation coefficient of head circumference among first- and second-born siblings was 0.343 (standard error 0.002) compared with 0.477 (standard error 0.002) for birthweight. These results were similar for later-born sibling pairs and only slightly influenced by other factors such as sex, fatherhood and time between pregnancies. Sensitivity analyses showed that substantial measurement error is required to explain the lower correlation for head circumference. In conclusion, the sibling correlation of head circumference was consistently weaker than that of birthweight. PMID- 14738545 TI - The Belgrade childhood diabetes study: prenatal and social associations for type 1 diabetes. AB - The aim of this study was to investigate whether prenatal and social factors are associated with the development of type 1 diabetes. A case-control study was conducted in Belgrade during the period 1994-97. A total of 105 children 41 weeks (11.11, [1.80, 68.52]); mother's consumption of foods containing nitrosamines during pregnancy (3.14, [1.68, 5.87]); and consumption of alcohol by father (4.54, [2.34, 8.79]). PMID- 14738546 TI - Risks of adverse pregnancy outcomes among Inuit and North American Indian women in Quebec, 1985-97. AB - We used Statistics Canada's linked stillbirth, live birth and infant death files to assess the risks of adverse pregnancy outcomes among Inuit and North American Indian vs. other ethnic women in Quebec, 1985-97 (1 125 462 singleton births). Mother tongue was used to define ethnicity, with the largest French language group as the reference. Main outcome measures are adjusted odds ratios (AOR) for preterm birth, small-for-gestational-age (SGA), stillbirth, neonatal and postneonatal death controlled for maternal age, education, marital status, parity, infant sex, community size, and community-level random effects using multilevel logit models. Inuit women had higher risks of preterm birth (AOR = 1.49, 95% CI [1.25, 1.78]) and immaturity-related infant mortality (AOR = 3.03 [1.36, 6.74]), while Indian women did not. Infants of Inuit (AOR = 0.39 [0.31, 0.49]) and Indian (AOR = 0.27 [0.24, 0.31]) women had substantially lower risks of SGA. Elevated risks of stillbirth were observed among Indian women [AOR = 1.53 (1.09, 2.15)], and of postneonatal death among both Inuit (AOR = 4.45 [2.74, 7.22]) and Indian (AOR = 1.86 [1.28, 2.70]) infants. Both Inuit and Indian infants had much higher risks of sudden infant death syndrome (SIDS) and infection-related mortality. Although the absolute risks of adverse outcomes declined from 1985-87 to 1995-97, the relative disparities between aboriginal and non-aboriginal women changed little over this period. We conclude that Inuit and Indian women have different risk profiles for adverse pregnancy outcomes, and that prevention of preterm birth among Inuit women, and of SIDS and infection related infant mortality in both aboriginal groups, are important targets for future research and intervention. PMID- 14738547 TI - Racial differences in leading causes of infant death in the United States. AB - We used linked birth/infant death records of over 23 million singletons belonging to six birth cohorts (1989-91 and 1995-97) and examined changes in race differentials in the overall and cause-specific infant mortality risks across time in the United States. Results show that infant mortality declined for all races during the time period, with disproportionately greater declines among non Hispanic American Indians (AIs). Among the leading causes of infant death, declines in mortality from sudden infant death syndrome (SIDS), respiratory distress syndrome (RDS) and congenital anomalies contributed the most to the overall decline in infant mortality in the 1995-97 cohorts, compared with the 1989-91 cohorts. Disproportionately greater reductions in mortality resulting from SIDS and congenital anomalies led to more rapid mortality declines among non Hispanic AIs than for other races. There are disturbing findings that infants of almost every race experienced increases in mortality from newborn affected by maternal complications of pregnancy (maternal complications) and that none of the race groups experienced a significant decline in mortality from disorders resulting from short gestation/low birthweight. PMID- 14738548 TI - Growth, development and health from early fetal life until young adulthood: the Generation R Study. AB - In this paper the Generation R Study is presented. This study examines growth, development and health in urban children from fetal life until young adulthood. With an integrated approach of epidemiological, clinical and basic research, it focuses on four primary areas of research: (1) growth and physical development; (2) behavioural and cognitive development; (3) diseases in childhood; and (4) health and health care for pregnant women and children. The general aims of the study are: 1. to describe normal and abnormal growth, development and health from fetal life until young adulthood in a multiethnic population-based cohort; 2. to identify biological, social and environmental determinants of normal and abnormal growth, development and health from fetal life until adulthood; 3. to examine the utilisation and effectiveness of current strategies for prevention and early identification of groups at risk. Eventually, this study will contribute to the development of strategies for optimising health and health care for pregnant women and children. The Generation R Study is a prospective population-based cohort study in Rotterdam, the Netherlands. In this urban setting, 10 000 children will be examined from early fetal life until young adulthood. Data are collected by physical examinations, questionnaires, interviews, ultrasound and biological samples. The study entered its pilot phase to test the logistics in December 2001. Full participant recruitment and complete data collection started in 2002. PMID- 14738549 TI - Repeated validation of parental self-reported smoking during pregnancy and infancy: a prospective cohort study of infants at high risk for allergy development. AB - Exposure to environmental tobacco smoke (ETS) during fetal life and infancy is closely related to the smoking habits of the parents. Estimates of exposure to ETS require valid and detailed information on changes in cigarette smoking over time. The objective was to test the validity of self-reported smoking among parents during pregnancy and early childhood in a cohort of children at high risk for allergy development by measurement of exhaled carbon monoxide (CO). The cohort comprised 117 families enrolled from the general population of pregnant women at admission to antenatal care. Data on parental tobacco smoking were obtained by interview and exhaled CO was measured (Micro-Smokerlyzer(R)) in parents twice during pregnancy and when the child was 6 and 18 months old. The median (range) exhaled CO levels were 3 (0-10) parts per million (ppm) for non smokers and 15 (1-39) ppm for smokers (P < 0.0005). A receiver operating characteristic (ROC) analysis was performed at each examination. The areas under the ROC curve were high for both mothers (between 0.88 and 0.99) and fathers (between 0.87 and 0.89), indicating exhaled CO as a good diagnostic tool for determining smoking status. Comparing the ROC areas obtained for mothers from late pregnancy and during infancy with the area from early pregnancy showed no statistical differences (P = 0.21, 0.43 and 0.44 respectively) and the same was true for fathers during infancy (P = 0.81). The level of 8 ppm was used as the cut-off between smokers and non-smokers, based on data from a pilot study. Using CO as a diagnostic tool for smoker status showed very high specificity (between 97 and 100%), indicating that very few persons claiming to be non-smokers had CO levels higher than 8 ppm. In conclusion, the validity of interview-obtained self reported smoking among parents during pregnancy and early childhood was high. Repeated interviews and CO measurements in a prospective study design did not change the validity, indicating a low risk of information bias. A structured interview combined with measurement of exhaled CO is a valid and reliable method for estimating ETS exposure to the fetus and young infant. PMID- 14738550 TI - Inter-observer reliability of radiological signs of necrotising enterocolitis in a population of high-risk newborns. AB - Radiological examination of the abdomen is critical to the diagnosis of necrotising enterocolitis (NEC). Previous studies on the reproducibility of radiological findings have been limited in size and based only on infants with the disease. We conducted a study among a sample of high-risk infants with and without the diagnosis of NEC: (1) to measure the degree of inter-observer agreement of NEC radiological diagnosis and signs and (2) identify a profile of radiological signs which led the observers to make a diagnosis of NEC. We collected 297 X-rays from a sample of 57 newborns admitted in 1999 to neonatal intensive care and neonatal surgery units in Rome, Italy. Three specialists in paediatric radiology examined the films independently and without any clinical information about patients. The analyses were conducted on a total of 891 forms filled in by the observers. Kappa values were calculated to measure the inter observer reliability. To identify the profiles of radiological signs, a multidimensional analysis, binary segmentation, was carried out. The reproducibility of radiographic signs was 0.55 (P < 0.01) for diffuse gaseous intestinal distention, 0.22 (P < 0.01) for bowel wall thickening, 0.10 (P < 0.01) for presence of portal venous gas and 0.29 (P < 0.01) for pneumatosis intestinalis. The agreement for radiographic diagnosis suspected/confirmed of NEC was 0.31 (P < 0.01). Among the 23 possible combinations of radiographic signs, the three radiologists indicated four profiles that produced a diagnosis of NEC containing, respectively, two, three, four and five signs. Our study found, in a large sample of radiographs selected from a population of infants with and without NEC, a poor reliability for NEC diagnosis and individual radiological signs among three expert radiologists. Clinical information and the presence of more than one radiological sign can reduce the margin of observer's error that inevitably exists when dealing with a diagnosis as difficult as NEC. PMID- 14738551 TI - New evidence for the theory of the stork. AB - Data from Berlin (Germany) show a significant correlation between the increase in the stork population around the city and the increase in deliveries outside city hospitals (out-of-hospital deliveries). However, there is no correlation between deliveries in hospital buildings (clinical deliveries) and the stork population. The decline in the number of pairs of storks in the German state of Lower Saxony between 1970 and 1985 correlated with the decrease of deliveries in that area. The nearly constant number of deliveries from 1985 to 1995 was associated with an unchanged stork population (no statistical significance). However, the relevance of the stork for the birth rate in that part of Germany remains unclear, because the number of out-of-hospital deliveries in this area is not well documented. A lack of statistical information on out-of-hospital deliveries in general is a severe handicap for further proof for the Theory of the Stork. PMID- 14738553 TI - Consensus conference on chronic viral hepatitis and HIV infection: updated Spanish recommendations. AB - Chronic hepatitis B and C represent a leading cause of morbidity and mortality among human immunodeficiency virus (HIV)-infected patients worldwide. New treatment options against both hepatitis B (HBV) and C (HCV) viruses have prompted us to update previous recommendations for the management of coinfected individuals. Fifteen topics (nine related to HCV, five to HBV and one to both viruses) were selected for this purpose. A panel of Spanish experts in the field was invited to review these areas and propose specific recommendations, which were scored according to the Infectious Disease Society of America (IDSA) grading system. These guidelines represent a comprehensive and updated overview on the management of hepatitis B and C in HIV-infected patients. PMID- 14738554 TI - DNA immunization encoding the secreted nonstructural protein 3 (NS3) of hepatitis C virus and enhancing the Th1 type immune response. AB - To induce a sustained and specific cellular immune response to hepatitis C virus (HCV), DNA immunization of mice was performed using plasmids containing the HCV nonstructural gene 3 (HCV/NS3). Plasmids were constructed such that the NS3 gene was expressed in a secreted form, a nonsecreted form or as a membrane-bound antigen. The plasmid encoding the secreted antigen induced the strongest humoral and cellular immunity and favoured the T-helper type 1 (Th1) pathway as shown by cytokine profiles and switching of antibody subclasses. Our study indicates that DNA immunization with a secreted form of HCV/NS3 is an effective means of inducing primary Th1 immune responses in the murine model. PMID- 14738555 TI - Detection and quantitation of HCV core protein in single hepatocytes by means of laser capture microdissection and enzyme-linked immunosorbent assay. AB - Immunohistochemistry provides valuable information concerning the localization and distribution of hepatitis C virus (HCV)-related proteins in histological sections of liver tissue, but does not readily permit their quantitation in individual cells and the staining intensity of cell immunodeposits cannot be calibrated with the current number of antigen molecules. We specifically detected and quantitated HCV core protein in single hepatocytes by coupling laser capture microdissection (LCM) with a sensitive enzyme-linked immunosorbent assay (ELISA). Quantitation of HCV core protein per cell was carried out on liver tissue cells obtained by LCM from fixed and stained frozen sections of 10 HCV-positive patients with chronic active hepatitis (CAH). Macromolecules from captured cells were solubilized in an extraction buffer and directly assayed for core protein using a sandwich ELISA. Calibration was achieved by developing a standard curve based on known concentrations of HCV core protein. Precision, linearity and sensitivity were verified for known numbers of microdissected tissue cells. In this study, the concentration of HCV core protein in single hepatocytes ranged from 7 to 56 pg/cell. Specificity was verified on 10 replicates of 10 HCV negative liver tissues. Immunohistochemical staining of HCV core protein was compared with the results of the soluble immunoassay for the adjacent liver tissue sections. Independent scoring of HCV immunostaining failed to parallel the LCM quantitative immunoassay. LCM-based immunoassay significantly expands our ability to investigate function-related antigens in apparently pure cell populations in HCV infection. PMID- 14738556 TI - Effect of interferon alpha and cell cycle progression on translation mediated by the hepatitis C virus 5' untranslated region: a study using a transgenic mouse model. AB - The effect of interferon alpha (IFN alpha) and the progression of the cell cycle on translation mediated by the 5' untranslated region (5'UTR) of hepatitis C virus (HCV) was evaluated in a transgenic mouse model containing the beta galactosidase (beta-gal) gene under the control of the mouse albumin promoter and HCV 5'UTR. The transgene was exclusively expressed in the liver and specifically in hepatocytes around the periportal area. IFN alpha significantly suppressed the expression of both the beta-gal gene product and its enzymatic activity at 6 h after the treatment of the mice. The mRNA level of the transgene and endogenous albumin gene expression were not affected, so this suppression was considered to be specific to 5'UTR-directed translation. Phosphorylation of the Stat1 protein was observed in the liver extract 20 min after the treatment, thus confirming a specific known effect of IFN alpha in vivo. We suggest that suppression of 5'UTR directed translation may be one of the mechanisms whereby IFN alpha exerts its anti-viral activity. We further investigated whether the restriction of 5'UTR directed translation in periportal hepatocytes may be explained by the proliferative state of the cell. Transgene expression was slightly enhanced in the liver 48 h after partial hepatectomy when a substantial number of hepatocytes entered cell cycle progression. However, 5'UTR-directed translation could not be detected in hepatocellular carcinoma lesions in transgenic mice that were induced to develop such tumours. We suggest that the state of differentiation of the cell, and not its proliferative capacity, is important for supporting HCV expression. This animal model may be a useful tool to dissect the control of HCV expression and to search for ways to block viral replication. PMID- 14738557 TI - Subtype mutations in the envelope 2 region including phosphorylation homology domain of hepatitis C virus do not predict effectiveness of antiviral therapy. AB - The aim of this study was to determine whether specific sequences of the phosphorylation homology domain (PePHD) region could be correlated with differences in response to antiviral therapy in patients infected with hepatitis C virus subtypes 1b, 2c, 3a and 4c/d. We included 43 patients (22 sustained responders and 21 nonresponders or relapsers) in the study, who were classified according to early viral decline during the first weeks of antiviral treatment and response at end of follow up. Type of mutations, mutation frequency, genetic diversity and phylogenetic relationships were compared at the PePHD and flanking regions. Phylogenetic trees showed that each sequence clustered together with those of the same subtype. Sequences from subtypes 1b and 4c/d resembled more closely the phosphorylation sites of protein kinase R and eIF2 alpha than sequences from genotypes 2c and 3a, the latter with higher response rates to interferon-alpha (IFN alpha) treatment. However, within specific subtypes, no separate clusters of responders and nonresponders were observed either at the beginning or at the end of follow up. We were not able to find any particular sequence or mutation in the PePHD region or in any other subregion of the fragment studied that allowed prediction of treatment response. PMID- 14738558 TI - High hepatitis B virus (HBV) DNA viral load is an important risk factor for HBV reactivation in breast cancer patients undergoing cytotoxic chemotherapy. AB - Hepatitis B virus (HBV) reactivation during cytotoxic chemotherapy for cancer may complicate treatment and cause liver damage. The complication has been reported to occur in 10% to over 50% of HBV carriers, but the factors that determine which patients will develop reactivation remain unclear. The objective of the study is to test the hypothesis that the prechemotherapy HBV DNA level is a risk factor for the development of HBV reactivation. We studied 41 women undergoing cytotoxic chemotherapy for breast cancer, 17 of whom developed reactivation and 24 who did not. We developed a novel, ultra-sensitive, real-time polymerase chain reaction assay for the measurement of HBV DNA. The sera of 37 patients (16 who developed reactivation and 21 who did not) were available for measurement of HBV DNA using this technique. The results showed that patients in the reactivation group had a significantly higher median HBV DNA load (1.03 x 10(6) copies/mL; range <2.9 x 10(3) to 8.723 x 10(7)) than did the nonreactivation group (<2.9 x 10(3) copies/ml; range <2.9 x 10(3) to 6.331 x 10(7)) (P < 0.001). The optimal cut-off between the two groups was found to be at serum HBV DNA level of 3 x 10(5), which gave a sensitivity of 81.0% and a specificity of 85.0%. In conclusion, for breast cancer patients receiving standard cytotoxic chemotherapy, a high HBV viral load prior to the administration of cytotoxic chemotherapy is a significant predictive factor for the development of HBV reactivation. Such information may be useful in determining which patients would benefit most from prophylactic antiviral therapy during cytotoxic chemotherapy. PMID- 14738559 TI - Interferon alfa2a induction therapy in combination with ribavirin and amantadine for the treatment of naive patients with chronic HCV infection. AB - Pilot studies have suggested that the addition of amantadine to interferon (IFN) is effective against hepatitis C virus (HCV). Furthermore, IFN induction therapy seems to improve virological response rates. In this open, randomized, multicentre trial we compared safety and efficacy of a triple therapy comprising IFN alpha 2a, ribavirin and amantadine using high induction doses (6 MU IFN alpha daily for the first 6 weeks) against a therapy with standard IFN alpha dosages over the entire treatment period plus amantadine and ribavirin. A total of 158 naive patients with chronic HCV infection were randomized 1:1. Group A (n = 81): induction therapy with 6 MU IFN alpha daily for 6 weeks, followed by 6 MU three times a week (tiw) for 18 weeks and then 3 MU tiw until week 48. Group B (n = 77): standard therapy with 6 MU IFN alpha tiw for 24 weeks, followed by 3 MU until week 48. All patients received oral ribavirin (10 mg/kg/day) and amantadine (200 mg/day). The triple therapy was safe and well tolerated. There were no significant differences between the groups with respect to biochemical response rates. Groups A and B did not differ in virological response rates at the end of treatment (33%vs 35%) or at the end of the 6 month follow up period (37%vs 39%). We could not detect favourable effects on sustained virological response rates using induction therapy, in either genotype 1 or non-1 infected patients. In summary, induction therapy with 6 MU IFN alpha daily did not result in increased overall response rates compared with standard IFN alpha dosages of 6 MU tiw. PMID- 14738560 TI - Thymosin-alpha 1 plus interferon-alpha for naive patients with chronic hepatitis C: results of a randomized controlled pilot trial. AB - In this pilot study, we evaluated the efficacy of interferon-alpha (IFN) plus Thymosin-alpha 1 (TA1) to that of IFN alone in naive patients with chronic hepatitis C. Twenty-two patients were randomized to receive interferon-alpha 2b (3 million units three times a week) plus thymosin-alpha l (900 microg/m2 body surface area) and 19 received interferon-alpha 2b alone at the same dose. Patients were treated for 6 months and followed up for another 6 months. Biochemical (alanine aminotransferase values) and virological (hepatitis C virus RNA) responses to treatment were determined. Combination treatment showed significantly higher efficacy than monotherapy in achieving virological end-of treatment response (P = 0.03). At 6-month follow up, the sustained biochemical and virological response was not different between the two groups. Our results indicate that the immune modulator TA1 may enhance the end-of-treatment response in naive patients with chronic hepatitis C. Higher doses and/ore more prolonged courses as well as the association with new interferon formulation such as pegylated interferons could improve the sustained response rates to this treatment. PMID- 14738562 TI - Evidence that plasma concentration rather than dose per kilogram body weight predicts ribavirin-induced anaemia. AB - Ribavirin in combination with interferon alpha-2 or pegylated interferon is the standard treatment for chronic hepatitis C. The current dosage recommendations for ribavirin are based on body weight (bw). Ribavirin is mainly eliminated by the kidneys and we have recently shown that ribavirin plasma concentrations are determined primarily by renal function. It is therefore reasonable to hypothesize that side-effects of ribavirin, i.e. anaemia, should be more closely related to plasma concentrations of ribavirin than to the dose per kg bw. A total of 108 consecutive patients eligible for treatment of chronic hepatitis C were studied. Ribavirin concentrations in plasma were measured by high-performance liquid chromatography (HPLC)-UV after solid-phase extraction in trough samples taken 4, 8 and 12 weeks after the treatment commenced. A total of 213 samples were obtained and the change in the haemoglobin level and the creatinine concentration was measured in addition to ribavirin. The dose of ribavirin per kg bw did not correlate with the drop in haemoglobin level induced by ribavirin. The concentration of ribavirin was non-linearly related to the drop in the haemoglobin level as revealed by fitting a standard Hill equation type dose response curve. The half maximal drop in haemoglobin was obtained at 4.4 microm. The results from this study suggest that the anaemia induced by ribavirin depends primarily on the concentration of ribavirin, and not on the dose per kg bw. This lends further support to the idea that ribavirin should be dosed according to renal function. PMID- 14738561 TI - Evolution of hepatitis B viral load and viral genome sequence during adefovir dipivoxil therapy. AB - Phase II and III clinical trials of adefovir dipivoxil (ADV) for the treatment of chronic hepatitis B have shown that this hepadnavirus polymerase inhibitor is well tolerated and effectively suppresses hepatitis B virus (HBV) replication. We therefore analysed the evolution of viral load and the emergence of HBV polymerase mutants in a 22-patient subgroup from a phase III clinical trial of ADV for the treatment of HBeAg-positive chronic hepatitis B. HBV DNA serum titres were quantified using a real-time polymerase chain reaction (PCR) assay with molecular hybridization probes. Emergence of polymerase mutants was assessed by direct sequencing of the viral reverse transcriptase domain after PCR amplification of HBV DNA isolated from serum. Our results indicated that ADV therapy effectively suppressed HBV replication in these patients (median serum HBV decrease at week 48 of treatment = 4.3 log10 copies/mL). The initial drop of HBV DNA titres in serum at week 12 of ADV therapy seemed to be predictive of subsequent HBe seroconversion (P = 0.059). Neither viral breakthrough nor the selection of drug resistant mutants were observed during the study period. Our results showed that ADV administration for 48-72 weeks effectively suppresses HBV replication without the emergence of resistant viral mutants. PMID- 14738563 TI - Hepatitis B surface antigenaemia following vaccination with a combined vaccine against hepatitis A and B. AB - We report a case of transient hepatitis B surface antigenaemia (HBsAg) following vaccination with a combined vaccine against hepatitis A and B in healthy adults. This phenomenon has been observed following administration of recombinant hepatitis B (monovalent) vaccine, mainly in newborns or dialysis patients. Reports on healthy adults are much less frequent and mostly concern blood donors. The frequency of its occurrence is largely unknown but its duration does not exceed 28 days. It is not detected by all available assays. It is caused by a passive transfer of antigen by vaccination, and not by viral replication; hence there is no risk for vaccination-induced infection. An important implication resulting from our findings is that the results of HBsAg assays should be interpreted according to the time elapsed since the last administration of a recombinant monovalent vaccine against hepatitis B or a combined vaccine against hepatitis A and B. PMID- 14738564 TI - The severity of liver fibrosis is associated with high leptin levels in chronic hepatitis C. AB - Recent attention has focused on the liver profibrogenic role of leptin in animal models. The purpose of this study was to evaluate the role of leptin and TNF alpha in the severity of liver fibrosis in patients with chronic hepatitis C (CHC). We used a radioimmunoassay to determine serum leptin concentrations in 77 consecutive patients with CHC and 22 healthy controls. Leptin was correlated with liver histological (METAVIR) and metabolic indices. Sixty five patients had none to moderate liver fibrosis (F0-F2) and twelve severe fibrosis (F3-F4). Steatosis was observed in all but 27 patients. Leptin was significantly increased in patients compared with controls and was significantly more elevated in females both in patients and controls. The age, age at infection, prothrombin index, body mass index (BMI), triglycerides, glycaemia, ferritin, leptin and TNF-alpha, were associated with severe fibrosis. Steatosis was significantly more pronounced in patients with severe than those without or moderate fibrosis (P = 0.04). Only leptin was significantly and independently associated with severe fibrosis (OR = 1.2, CI 95%: 1.1-1.4, P = 0.03). Leptin was significantly associated with BMI (r = 0.64, P < 0.001) and glycaemia (r = 0.43, P < 0.001). Significant correlations were found between steatosis and BMI (r = 0.30, P < 0.01) and glycaemia (r = 0.30, P < 0.01). In patients with CHC and higher BMI and glycaemia levels, the severity of liver fibrosis is associated with serum leptin. TNF-alpha is a putative candidate involved in the mechanism. PMID- 14738565 TI - Human cell-derived microparticles promote thrombus formation in vivo in a tissue factor-dependent manner. AB - BACKGROUND: Circulating microparticles of various cell types are present in healthy individuals and, in varying numbers and antigenic composition, in various disease states. To what extent these microparticles contribute to coagulation in vivo is unknown. OBJECTIVES: To examine the in vivo thrombogenicity of human microparticles. METHODS: Microparticles were isolated from pericardial blood of cardiac surgery patients and venous blood of healthy individuals. Their numbers, cellular source, and tissue factor (TF) exposure were determined using flow cytometry. Their in vitro procoagulant properties were studied in a fibrin generation test, and their in vivo thrombogenicity in a rat model. RESULTS: The total number of microparticles did not differ between pericardial samples and samples from healthy individuals (P = 0.786). In both groups, microparticles from platelets, erythrocytes, and granulocytes exposed TF. Microparticle-exposed TF antigen levels were higher in pericardial compared with healthy individual samples (P = 0.036). Pericardial microparticles were strongly procoagulant in vitro and highly thrombogenic in a venous stasis thrombosis model in rats, whereas microparticles from healthy individuals were not [thrombus weights 24.8 (12.2-41.3) mg vs. 0 (0-24.3) mg median and range; P < 0.001]. Preincubation of pericardial microparticles with an inhibitory antibody against human TF abolished their thrombogenicity [0 (0-4.4) mg; P < 0.01], while a control antibody had no effect [19.6 (12.6-53.7) mg; P > 0.05]. The thrombogenicity of the microparticles correlated strongly with their TF exposure (r = 0.9524, P = 0.001). CONCLUSIONS: Human cell-derived microparticles promote thrombus formation in vivo in a TF dependent manner. They might be the direct cause of an increased thromboembolic tendency in various patient groups. PMID- 14738566 TI - The validity of the MacNew Quality of Life in heart disease questionnaire. AB - BACKGROUND: A previous review suggested that the MacNew Quality of Life Questionnaire was the most appropriate disease-specific measure of health-related quality of life among people with ischaemic heart disease. However, there is ambiguity about the allocation of items to the three factors underlying the MacNew and the factor structure has not been confirmed previously among the people in the UK. METHODS: The MacNew Questionnaire and the SF-36 were administered to 117 newly admitted patients to a tertiary referral centre in Northern Ireland. All patients had been diagnosed with ischaemic heart disease. RESULTS: A confirmatory factor analysis was conducted on the factor structure of the MacNew and the model was found to be an inadequate fit of the data. A quantitative and qualitative analysis of the items suggested that a five factor solution was more appropriate and this was validated by confirmatory factor analysis. This new structure also displayed strong evidence of concurrent validity when compared to the SF-36. CONCLUSION: We recommend that researchers should submit scores obtained from items on the MacNew to secondary analyses after being grouped according to the factor structure proposed in this paper, in order to explore further the most appropriate grouping of items. PMID- 14738567 TI - The mRNA expression of hTERT in human breast carcinomas correlates with VEGF expression. AB - BACKGROUND: Telomerase is a ribonucleoprotein enzyme that synthesises telomeres after cell division and maintains chromosomal stability leading to cellular immortalisation. hTERT (human telomerase reverse transcriptase) is the rate limiting determinant of telomerase reactivation. Telomerase has been associated with negative prognostic indicators in some studies. The present study aims to detect any correlation between hTERT and the negative prognostic indicators VEGF and PCNA by quantitatively measuring the mRNA expression of these genes in human breast cancer and in adjacent non-cancerous tissue (ANCT). MATERIALS AND METHODS: RNA was extracted from 38 breast carcinomas and 40 ANCT. hTERT and VEGF165, VEGF189 and PCNA mRNA expressions were estimated by reverse transcriptase-PCR (RT PCR) and Taqman methodology. RESULTS: The level of expression of VEGF-165 and PCNA was significantly higher in carcinoma tissue than ANCT (p = 0.02). The ratio of VEGF165/189 expression was significantly higher in breast carcinoma than ANCT (p = 0.025). hTERT mRNA expression correlated with VEGF-189 mRNA (p = 0.008) and VEGF165 (p = 0.07). CONCLUSIONS: hTERT mRNA expression is associated with the expression of the VEGF189 and 165 isoforms. This could explain the poorer prognosis reported in breast tumours expressing high levels of hTERT. The relative expression of the VEGF isoforms is significantly different in breast tumour to ANCT, and this may be important in breast carcinogenesis. PMID- 14738569 TI - Antibody responses to Brugia malayi antigens induced by DNA vaccination. AB - BACKGROUND: DNA vaccination is a convenient means of immunizing animals with recombinant parasite antigens. DNA delivery methods are believed to affect the qualitative nature of immune responses to DNA vaccines in ways that may affect their protective activity. However, relatively few studies have directly compared immune responses to plasmids encoding the same antigens after injection by different routes. Therefore, the purpose of this study was to explore the influence of the route of administration on antibody responses to plasmids encoding antigens from the filarial nematode parasite Brugia malayi. METHODS: Four B. malayi genes and partial genes encoding paramyosin (BM5), heat shock protein (BMHSP-70), intermediate filament (BMIF) and a serodiagnostic antigen (BM14) were inserted in eukaryotic expression vectors (pJW4303 and pCR trade mark 3.1). BALB/c mice were immunized with individual recombinant plasmids or with a cocktail of all four plasmids by intramuscular injection (IM) or by gene gun intradermal inoculation (GG). Antibody responses to recombinant antigens were measured by ELISA. Mean IgG1 to IgG2a antibody ratios were used as an indicator of Th1 or Th2 bias in immune responses induced with particular antigens by IM or GG immunization. The statistical significance of group differences in antibody responses was assessed by the non-parametric Kruskal-Wallis test. RESULTS: Mice produced antibody responses to all four filarial antigens after DNA vaccination by either the IM or GG route. Antibody responses to BM5 paramyosin were strongly biased toward IgG1 with lower levels of IgG2a after GG vaccination, while IM vaccination produced dominant IgG2a antibody responses. Antibody responses were biased toward IgG1 after both IM and GG immunization with BMIF, but antibodies were biased toward IgG2a after IM and GG vaccination with BMHSP-70 and BM14. Animals injected with a mixture of four recombinant plasmid DNAs produced antibodies to all four antigens. CONCLUSIONS: Our results show that monovalent and polyvalent DNA vaccination successfully induced antibody responses to a variety of filarial antigens. However, antibody responses to different antigens varied in magnitude and with respect to isotype bias. The isotype bias of antibody responses following DNA vaccination can be affected by route of administration and by intrinsic characteristics of individual antigens. PMID- 14738568 TI - Expression of the pro-angiogenic factors vascular endothelial growth factor and interleukin-8/CXCL8 by human breast carcinomas is responsive to nutrient deprivation and endoplasmic reticulum stress. AB - BACKGROUND: The expression of pro-angiogenic cytokines, such as vascular endothelial growth factor (VEGF) and interleukin-8/CXCL8 (IL-8), plays an important role in tumor growth and metastasis. Low oxygen tension within poorly vascularized tumors is thought to be the prime stimulus causing the secretion of VEGF. The expression of IL-8 by solid tumors is thought to be primarily due to intrinsic influences, such as constitutive activation of nuclear factor kappa B (NF-kappaB). However, VEGF expression is responsive to glucose deprivation, suggesting that low concentrations of nutrients other than oxygen may play a role in triggering the pro-angiogenic phenotype. Glucose deprivation causes endoplasmic reticulum (ER) stress and alters gene expression through the unfolded protein response (UPR) signaling pathway. A branch of the UPR, known as the ER overload response (EOR), can cause NF-kappaB activation. Thus, we hypothesized that treatments that cause ER stress and deprivation of other nutrients, such as amino acids, would trigger the expression of angiogenic cytokines by breast cancer cell lines. RESULTS: We found that glutamine deprivation and treatment with a chemical inducer of ER stress (tunicamycin) caused a marked induction of the secretion of both VEGF and IL-8 protein by a human breast adenocarcinoma cell line (TSE cells). Glutamine deprivation, glucose deprivation and several chemical inducers of ER stress increased VEGF and IL-8 mRNA expression in TSE and other breast cancer cell lines cultured under both normoxic and hypoxic conditions, though hypoxia generally diminished the effects of glucose deprivation. Of all amino acids tested, ambient glutamine availability had the largest effect on VEGF and IL-8 mRNA expression. The induction of VEGF mRNA expression, but not IL-8, was sustained and closely corresponded with the upregulated expression of the ER stress-responsive genes glucose-regulated protein 78 (GRP78) and growth arrest and DNA damage inducible gene 153 (GADD153). CONCLUSION: These results suggest that nutrient deprivation within the solid tumor microenvironment might contribute to the activation of a pro-angiogenic phenotype. The angiogenic switch may act to increase blood supply in response to nutrient deprivation as well as hypoxia. PMID- 14738570 TI - Fas induces apoptosis in human coronary artery endothelial cells in vitro. AB - BACKGROUND: Published work suggests that some types of endothelial cells undergo apoptosis in response to ligation of the receptor Fas (CD95, APO1) but other types are resistant. Because heterogeneity among endothelial cells from different tissues, has been demonstrated, the purpose of this study was to determine, if Fas ligation and/or activation by human Fas ligand induces apoptosis and caspase activities, in cultured human coronary artery endothelial cells, and the differences between TNF-a and FAS induced apoptosis in these cells. RESULTS: Cultured human coronary artery endothelial cells (HCAEC) were exposed to the monoclonal Fas-activating antibody CH-11, to purified recombinant human Fas ligand, to the Fas-neutralizing antibody ZB4, or to purified recombinant human TNF-alpha. Apoptosis was detected by assessment of chromatin condensation and nuclear fragmentation and by assay of the enzymatic activities of Caspase 1 and Caspase 3 with membrane-permeable substrates applied to intact cells. Fas protein was detected by immunoblotting of HCAEC lysates. Apoptosis was induced in HCAEC by purified Fas ligand or by the monoclonal activating antibody CH-11 at concentrations of 25 or 200 ng/ml, but not by nonspecific isotype-matched immunoglobulins. The apoptotic index elicited by either Fas activator was equal to that induced by TNF-a (3.0-3.6-fold versus control, p < 0.01). The Fas neutralizing antibody ZB4 abrogated HCAEC apoptosis induced by CH-11, but had no inhibitory effect on apoptosis in response to TNF-a. Fas ligation significantly increased the activities of both Caspase 1 and Caspase 3 at 20 hours of stimulation (1.7- and 2.0-fold versus control, both p < 0.05); in contrast, purified TNF-a increased the activity of Caspase 3 but not Caspase 1 (2.1-fold, p < 0.05). Western blotting of HCAEC lysates with antibody CH-11 identified a single immunoreactive protein of 90 kDa. CONCLUSIONS: Cultured human coronary artery endothelial cells express functional Fas capable of inducing apoptosis in response to either purified Fas ligand or receptor-activating monoclonal antibodies, at levels equal to those inducible by purified TNF-alpha. Immunologic studies and differential kinetics of caspase activation suggest that Fas and TNF alpha induce apoptosis in HCAEC by signaling pathways that are distinct but equal in potency. PMID- 14738572 TI - Management of meningitis in resource-poor settings. PMID- 14738571 TI - Antibiotic treatment for bacterial meningitis in children in developing countries. AB - Bacterial meningitis causes 125,000 deaths each year in infants and young children and 96% of these occur in less developed countries where up to 50% of children with this disease die and 25-50% of survivors have neurological sequelae. Although 3rd-generation cephalosporins are optimal empirical therapy for bacterial meningitis, they are unaffordable in many developing countries. The majority of children worldwide are currently treated with cheaper alternatives. This paper reviews the challenges facing clinicians treating bacterial meningitis in developing countries, highlighting the problem of changing patterns of antibiotic resistance. In particular, it details the evidence for the use of chloramphenicol and 3rd-generation cephalosporins. PMID- 14738573 TI - Pyrexia of unknown origin in children: a review of 102 patients from Turkey. AB - Pyrexia of unknown origin (PUO) has not been appropriately investigated in Turkish children and therefore a study was undertaken to determine the causes of PUO and to evaluate which clinical procedures are useful in establishing a diagnosis. A total of 102 children fitting the classical PUO criteria seen in our clinic between 1995 and 2002 were investigated retrospectively. Infections, collagen vascular disorders, malignancy and miscellaneous conditions constituted 44.2%, 6.8%, 11.7% and 24.5% of cases, respectively, while 12.8% of the cases remained undiagnosed. Enteric fever, brucellosis and respiratory tract infections were the most commonly encountered infections, whereas familial Mediterranean fever was the commonest non-infectious disorder. Biopsy, aspiration, serology, bacteriology, radiology and observation of the clinical course were the most useful diagnostic procedures. PMID- 14738574 TI - Leukaemia in children in Papua New Guinea: an unusual pattern. AB - We report data on 110 children aged <15 years diagnosed with leukaemia during two periods covering 13.25 years. The data sets were consistent. The reported incidence of leukaemia was low. Only 34 (31%) of the children were diagnosed with acute lymphoblastic leukaemia (ALL) compared with 54 (49%) children with acute myeloid leukaemia (AML). The overall mean (SD) age was 6.6 (3.5) years, 6.1 (3.5) for ALL and 6.9 (3.5) for AML. There was no evidence of an early childhood peak of ALL. The male : female ratio was 1.2 : 1 for all leukaemias, 1.3 for ALL and 1.25 for AML. Only eight (22%) of those diagnosed with ALL were classified as type L1. Our figures reflect a relative absence of the common (cALL) cell type in early childhood leukaemia and support the role of infection and its effect on the immune system in the aetiology of childhood leukaemia. Our data also revealed an unusually high proportion of chronic myeloid leukaemia (CML). PMID- 14738575 TI - Human immunodeficiency virus and urinary tract infections in children. AB - This was a retrospective study of HIV-infected children aged 0-12 years attending the King Edward VIII Hospital in Durban, South Africa over a 5-year period (January 1996 to December 2001) with culture-proven urinary tract infection (UTI). UTI was defined as the presence of a single bacterial growth of >10(5) colony-forming units/ml in a clean-catch, mid-stream urine sample or >10(3) organisms/ml in a catheter or suprapubic aspirate of urine. HIV/AIDS was diagnosed in accordance with World Health Organization and/or Centers for Disease Control criteria. Comparison between HIV-positive and HIV-negative children with UTI was done using the chi2 and Wilcoxon rank sum tests. Of the 55 children recruited into the study, 29 (52.1%) were HIV-positive and 26 (47.3%) HIV negative. Escherichia coli was isolated in 50 (87.2%) children. Clinical presentation, aetiological agents, response to therapy and renal function were similar in both groups. This study showed no significant impact of HIV/AIDS on the presentation of UTI in children. PMID- 14738576 TI - Immune response in HIV-1-infected children with thalassaemia given a primary course of DPT vaccine before acquiring HIV-1 infection. AB - The effect of HIV infection on immune response to diphtheria and tetanus primary immunisation was investigated in 24 HIV-1-positive multi-transfused (MT) children with thalassaemia and compared with 48 HIV-1-negative MT thalassaemic children and 36 HIV-1-negative non-transfused (NT) children in the community. Diphtheria and tetanus antibody levels in the HIV-1-positive MT group were comparable with the two HIV-negative groups. The proportions of children with antibody titres below the protective level (i.e. <0.01 IU/ml) for antidiphtheria antibodies were 20.8, 16.6 and 16.6%, and 12.5, 12.5 and 13.9% for anti-tetanus antibodies in the three groups, respectively. On the other hand, delayed-type hypersensitivity (DTH) response to diphtheria and tetanus antigens was significantly depressed in the HIV-1-positive group compared with the HIV-negative controls. The mean percentages of both mature (CD20+) and immature (CD10+) B-cell counts were significantly higher in the HIV-1-positive group than in the HIV-negative MT and NT groups (p<0.05). Levels of serum immunoglobulins and spontaneously secreted immunoglobulins were significantly higher in the HIV-1-positive group compared with both HIV-negative groups. The HIV-1-positive group showed a mean (SD) IL-6 of 52.9 (28.8) pg/ml compared with 23.7 (12.1) pg/ml and a detection rate of 54.2% in the HIV-negative MT group, and 23.6 (8.2) pg/ml and a 50% detection rate in the HIV-negative NT group. The IL-2 level was significantly lower (p<0.05) in the HIV-1-positive group [41.7% detection rate and mean (SD) 28.8 (17.1) pg/ml] than in the HIV-negative MT and NT groups [75% and 83.3% detection rates and mean (SD) 57.2 (42.3) pg/ml and 99.3 (51.1) pg/ml, respectively]. During follow-up for 3 years, the frequency of major infections was significantly higher in the HIV-1 positive group than in the other two groups. Acute pneumonia and acute sinusitis were the predominant infections regardless of HIV status while primary bacteraemia, osteomyelitis, pyogenic meningitis and septic arthritis were common in the HIV-1-positive group. We conclude that, in HIV-1-infected children pre immunised with DPT, DTH response to diphtheria and tetanus antigens might be more reliable than anti-diphtheria and anti-tetanus antibody levels in predicting susceptibility to major bacterial infections. PMID- 14738577 TI - Neonatal morbidity and mortality in a Tanzanian tertiary care referral hospital. AB - In developing countries, neonatal mortality accounts for 50-70% of infant mortality. The purpose of this study was to describe morbidity and mortality patterns, with a focus on neonatal infections, in a Tanzanian special care baby unit (SCBU). During a 3-month period, 246 consecutive admissions to the SCBU at Kilimanjaro Christian Medical Centre were audited. Prematurity, low birthweight and suspected infection accounted for 61% of all admissions. The overall mortality rate was 19%, but varied considerably according to gestational age, birthweight and diagnosis. Thirty-one neonates (two-thirds of all deaths) died during the 1st 24 hours of life. Of 27 infants admitted on grounds of perinatal asphyxia, 11 (41%) died, and, of 19 infants with a gestational age <31 weeks, 13 (68%) died. More than two-thirds of all infants were treated with antibiotics. Septicaemia confirmed by blood culture was found in 16 cases. The susceptibility pattern of bacterial isolates did not indicate high rates of resistance to commonly used antibacterial agents. A reduction in the number of preterm deliveries and improved perinatal care to avoid and treat perinatal asphyxia would be the two most important measures in reducing neonatal mortality in this setting. PMID- 14738578 TI - Werdnig-Hoffmann disease with congenital hypothyroidism. AB - Congenital hypothyroidism is often associated with other congenital anomalies. In some instances it is difficult to differentiate congenital hypothyroidism from Werdnig-Hoffmann's disease. We report a case of congenital hypothyroidism associated with Werdnig-Hoffmann's disease that appears to be the first of its kind in the literature. PMID- 14738579 TI - Gastric teratoma--a rare benign tumour of neonates. AB - We describe our experience of two neonates with gastric teratoma, one a low birthweight, premature baby who had a massive, immature teratoma. Complete excision is the appropriate treatment. PMID- 14738580 TI - Central nervous system involvement in neonatal rotavirus infection. AB - Rotavirus is an ubiquitous virus associated with severe gastro-enteritis in children under 2 years of age. However, serious conditions, including central nervous system involvement, have been associated with rotavirus infection. We investigated the clinical signs and symptoms of rotavirus infection in 156 neonates, specifically focusing on neurological signs. Neurological signs identified were apnoea, jitteriness, convulsions and poor sucking. Of the 156 babies, 66 were rotavirus-positive and 90 rotavirus-negative. In the rotavirus positive group, 19.7% manifested apnoea compared with 22.2% in the rotavirus negative group. The other three symptoms occurred in <10% of the neonates. Rotavirus dsRNA was detected by RT-PCR in the CSF of two. One had apnoea, the other no CNS symptoms. Although rotavirus infection in neonates is hardly ever asymptomatic and may cause death, it has not been associated with CNS symptomatology in our studies. PMID- 14738581 TI - A large cerebral hydatid cyst associated with liver cyst. AB - Hydatid disease in childhood is a serious health problem in Turkey and other places where the parasite is endemic. An 8-year-old girl from a rural area was admitted with headache, recurrent abdominal pain, abdominal distension and hypo aesthesia and paresis of the right arm. Computerised tomography (CT) scans demonstrated a large intracranial hydatid cyst. A hydatid cyst was detected in the liver also. Her mother had been treated previously for disseminated hydatid disease. The cerebral cyst was removed but only subtotal excision of the hepatic cyst was possible. She was treated with albendazole, has fully recovered and, after 12 months of follow-up, there is no sign of recurrence. PMID- 14738582 TI - Primary central nervous system Burkitt's lymphoma presenting with proptosis. PMID- 14738583 TI - Rational design of powder formulations for tamp filling processes. AB - Tamp filling processes are widely used for the filling of powders into hard gelatin capsules, whereby capsule fill weight is controlled by the formation of a loosely packed plug of material that is dispensed into the capsule shell. To rationally design formulations for tamp filling processes the formulator must have an intimate knowledge of the synergy between machine parameters and powder properties and the corresponding effect on product quality. However, despite ubiquitous use throughout the pharmaceutical industry, relatively little is understood about the design of powders for tamp filling processes. The aim of the following review is to summarize the published literature to date from a formulation design perspective and to provide a framework for future scientific research. PMID- 14738584 TI - In-vitro and in-vivo studies of cefpirom using bile salts as absorption enhancers. AB - Cephalosporins have to be administered by injection because of the poor intestinal absorption of the orally delivered drugs. Because of the obvious drawbacks of drug delivery by injection, the development of alternatives with enhanced oral bioavailability is receiving much attention in pharmaceutical research. Cefpirom (Cp) is a new semi-synthetic amino-2-thiazolyl-methoxyimino cephalosporin that has been substituted in position 3 with a cyclopenteno pyridinium group in order to create a zwitterionic compound. It exhibits highly hydrophilic properties, as shown from its extremely low partition coefficient, and therefore its lipophilicity was increased using bile salts. The effect of this on the partition coefficients determined in the n-octanol/buffer system was confirmed using an in-vitro transport model with artificial and biological membranes. The pharmacokinetic properties of Cp were investigated in rabbits after intraduodenal administration with and without bile salts. Furthermore, the physiological compatibility of the bile salts was investigated using active D glucose transport. PMID- 14738585 TI - Floating microspheres bearing acetohydroxamic acid for the treatment of Helicobacter pylori. AB - This investigation is part of our ongoing effort to develop effective drug delivery systems for the treatment of Helicobacter pylori infection using polycarbonate (PC) floating microspheres as drug carriers. In an effort to augment the anti-H. pylori effect of acetohydroxamic acid (AHA), floating PC microspheres, which have the ability to reside in the gastrointestinal (GI) tract for an extended period, were prepared by emulsion (O/W) solvent evaporation technique. The effect of PC concentration on the morphology, particle size, entrapment efficiency and drug release rate was studied. In-vitro studies confirmed the excellent floating properties of PC microspheres. In-vitro and in vivo growth inhibition studies were performed on developed system(s) taking isolated cultures of H. pylori and H. pylori-infected Mongolian gerbils, respectively. The drug and PC microspheres both showed anti-H. pylori activity in vivo, but the required dose of AHA was effectively reduced by a factor of 10 in the case of PC microspheres. In conclusion, the floating microspheres more effectively cleared H. pylori from the GI tract than the drug because of the prolonged gastric residence time resulting from the excellent buoyancy of the PC. PMID- 14738586 TI - Use of a cholesterol-rich microemulsion that binds to low-density lipoprotein receptors as vehicle for etoposide. AB - A cholesterol-rich microemulsion (LDE) that binds to low-density lipoprotein (LDL) receptors is selectively taken up by malignant cells that overexpress those receptors and may be used as vehicle for antineoplastic agents. This study aimed to develop the association of etoposide with LDE. It was firstly observed that etoposide poorly associates with the microemulsion, therefore the experiments were performed with a lipophilic fatty acid derivative of the drug. The association of etoposide oleate with LDE was almost 100% and was tested for physical and chemical stability, as well as for cellular uptake, toxicity in mice and cytotoxic activity against a neoplastic cell line (NCI-H292). Uptake and cytotoxic activity of LDE-etoposide oleate by NCI-H292 cells was mediated by LDL receptors. The anti-proliferative activity of LDE-etoposide oleate against the neoplastic cells was smaller than that of etoposide oleate (IC50 (drug concentration required to inhibit 50% of the cell growth) = 0.48 and 0.19 mM, respectively). This difference, however, can be ascribed to the activity of the commercially used vehicle and not the drug itself because when this vehicle was added to the cultures with LDE-etoposide oleate, the IC50 decreased. On the other hand, the tolerability of LDE-etoposide oleate to mice was remarkable, such that its lethal dose (LD50) was about five-fold that of the commercial formulation (LD50 = 315 and 58 mg kg(-1), respectively). In conclusion, LDE-etoposide oleate association is stable and the cytostatic activity of the drug is preserved while its toxicity to animals is small. By diminishing the side effects and directing etoposide to neoplastic tissues, LDE may be regarded as an advance in chemotherapy with this drug. PMID- 14738587 TI - Chitosan salts as nasal sustained delivery systems for peptidic drugs. AB - The aim of this study was to describe a sustained drug release system based on chitosan salts for vancomycin hydrochloride delivery. Chitosan lactate, chitosan aspartate, chitosan glutamate and chitosan hydrochloride were prepared by spray drying technique. Vancomycin hydrochloride was used as a model peptidic drug, the nasal sustained release of which should avoid first-pass metabolism in the liver. This in-vitro study evaluated the influence of chitosan salts on the release behaviour of vancomycin hydrochloride from the physical mixtures at pH 5.5 and 7.4. In-vitro release of vancomycin was retarded by chitosan salts and, in particular, chitosan hydrochloride provided the lowest release of vancomycin. PMID- 14738588 TI - Exposure of the yeast Candida albicans to the anti-neoplastic agent adriamycin increases the tolerance to amphotericin B. AB - Cancer patients experience a high incidence of fungal infections due to their immuno-suppressed condition. This work has investigated the interaction of an anti-neoplastic agent, adriamycin (doxorubicin), with the yeast Candida albicans and examined whether this drug altered the susceptibility of the yeast to amphotericin B - an anti-fungal agent used for the treatment of systemic fungal infections in cancer patients. Exposure to adriamycin for 24h increased the growth of C. albicans and increased the tolerance to amphotericin B by a small, but statistically significant, extent. Growth in adriamycin-supplemented medium suppressed the respiration rate of C. albicans, which resulted in a decrease in the ergosterol content of the fungal cell membrane. The tolerance to amphotericin B was lost after exposure to adriamycin for 48 h, which coincided with a restoration in the respiration rate and the ergosterol content of the fungal cell membrane. This work demonstrated that short-term exposure (24 h) to adriamycin increased the tolerance of C. albicans for amphotericin B, which may be mediated by a decrease in the ergosterol content as a result of an adriamycin-induced disruption of oxidative phosphorylation. PMID- 14738589 TI - Effect of physiological media on the stability of surface-adsorbed DNA-dendron gold nanoparticles. AB - Plasmid DNA was adsorbed onto 87-nm gold nanoparticles to which were adsorbed a layer of novel cationic dendrons. The behaviour of this DNA-dendron-gold system in cell culture media has been described. Adsorption onto the gold nanoparticles of lipophilic cationic dendrons, with either 8 [(C12)(3)Lys7(NH2)(8)] or 16 [(C12)(3)Lys15(NH2)(16)] free amino groups on their outer surfaces and incorporating a nuclear localization signal peptide (NLS), resulted in positively charged nanoparticles with a corresponding small increase in particle size. Evidence suggested that the interaction between the gold nanoparticles and the dendron was mediated by hydrophobic forces. With an increase in ionic strength, the apparent particle size of the dendron-stabilized-gold particles increased, but at higher salt concentrations than plain gold sols. Addition of plasmid DNA did not markedly reduce the surface potential of the dendron-gold complex but resulted in an approximately 10-20% increase in hydro-dynamic diameter. Increasing ionic strength increased the apparent size of the DNA-dendron-gold particles, up to a maximum diameter of approximately 900 nm. Importantly, in cell culture media the size of the DNA-dendron-gold nanoparticles increased markedly, as surface potential was reduced. The presence of serum components partially ameliorated these effects, possibly due to steric stabilization of the particles. Release of the DNA from the complex was compromised in cell culture media (compared with water). This, coupled with the flocculation of the carrier, demonstrated the importance of testing delivery systems in the presence of relevant physiologically based fluids before cell culture or in-vivo studies. PMID- 14738590 TI - Effect of ketoprofen and its enantiomers on the renal disposition of methotrexate in the isolated perfused rat kidney. AB - Non-steroidal anti-inflammatory drugs (NSAIDs) have been shown to inhibit the renal tubular secretion of methotrexate. However, the relative contribution of the active S- and inactive R-enantiomers is unknown. This study examined the effect of racemic ketoprofen and its enantiomers on the renal disposition of methotrexate in the isolated perfused rat kidney (IPK). Nineteen kidneys were divided between a control and three treatment groups. Controls were perfused with methotrexate alone (25 micrograms mL-1, n = 5) over three 30-min periods. Treatment groups were perfused with methotrexate (25 micrograms m-1) for the first period, followed by a second period of methotrexate (25 micrograms mL-1) plus R- (n = 5), S- (n = 5) or RS-ketoprofen (n = 4) at 25 micrograms mL-1, and a third period of methotrexate (25 micrograms mL-1) plus R-, S- or RS-ketoprofen (50 micrograms mL-1). Perfusate and urine were collected over 10-min intervals. Methotrexate was measured by HPLC and its binding in perfusate by ultrafiltration. The clearance ratio (CR) for methotrexate was obtained by dividing the renal clearance by the product of its fraction unbound and the glomerular filtration rate. During control experiments, there was no significant change in the CR over 90 min. R-, S- and RS-ketoprofen at 50 micrograms mL-1 reduced the CR of methotrexate significantly, but there was no difference between the three groups. While the enantiomers of ketoprofen reduced the renal excretion of methotrexate, the interaction was not enantioselective. PMID- 14738591 TI - Absorption, distribution and excretion of 14C-chloroquine after single oral administration in albino and pigmented rats: binding characteristics of chloroquine-related radioactivity to melanin in-vivo. AB - Chloroquine is an antimalarial agent that has been reported to have distinct affinity to melanin. After single oral administration of 14C-chloroquine at a dose of 20mg kg-1 under non-fasting conditions, the absorption, distribution and excretion of 14C-chloroquine-related radioactivity were studied in albino and pigmented rats. The objectives of the study were to investigate differences in the disposition of chloroquine between albino and pigmented rats and to define its in-vivo binding characteristics to melanin-containing ocular tissues. Extensive uptake of radioactivity into tissues was indicated by higher concentrations in most tissues compared with serum and there was no quantitative differences in the distribution of radioactivity found between albino and pigmented rats except for melanin-containing tissues, such as the uveal tract of the eye and perhaps hair follicles. There was selective and strong binding of drug-related compounds to these tissues in pigmented rats. The uveal tract concentrations reached the maximum value of 158.42 +/- 7.86 micrograms equiv g-1 (mean+/-s.e.) at 1 week and decreased very slowly with a terminal half life of 4476 h (187 day). The uveal tract concentrations at 24 weeks were still high (67.75 +/- 6.19 micrograms equiv g-1). The AUC for uveal tract was 842.3 mg.h g 1. A relatively high concentration was still determined in the uveal tract even at 48 weeks after single oral dosing by whole-body autoradiography. The uveal tracts separated from one eye of each rat were extracted with 0.067 M phosphate buffer (pH 7.4) and 1 M HCl-EtOH (30:70) successively. In pigmented rats, almost all radioactivity was released from the tissue with 1 M HCl-EtOH (30:70), indicating that the strong binding by melanin was reversible, and that hydrophobic or electrostatic interaction would play a critical role in the binding of chloroquine and its metabolites with the melanin-containing ocular tissues. Approximately 70% of the radioactivity given was recovered in urine and faeces up to 144 h after dosing both in pigmented and albino rats. The excretion pattern in pigmented rats was similar to that seen in albino rats. PMID- 14738592 TI - Free radical scavenging and antioxidative activity of melatonin derivatives. AB - This article describes the synthesis and antioxidative properties of melatonin derivatives. Tryptamines and cysteinyl or mercaptopropionyl derivatives were deliberately condensed with coupling reagents to give melatonin derivatives 4a-d and 5a, b. The preliminary evaluation indicated that compound 4c showed improved scavenging activity compared with vitamin C (IC50 43 microM vs 65 microM, where IC50 is the concentration of the test compound that induced a change of 50% in absorbance during the 30 min observation) on diphenyl-p-picrylhydrazyl (DPPH) tests. Derivative 5b, which possesses the thiolactyl moiety, showed moderate potency compared with melatonin (IC50 235 microM vs 690 microM) in the H(2)O(2) scavenging test. Intriguingly, 4c displayed 2-fold more potency than melatonin (IC50 51 microM vs 125 microM) in scavenging NO in the macrophage model. These results suggested that the cysteinyl-conjugated derivative 4c may be a suitable lead to further optimize potent antioxidants for certain oxidative stress conditions. PMID- 14738593 TI - Effect of methotrexate on Th1 and Th2 immune responses in mice. AB - We investigated the effect of the anti-rheumatic drug methotrexate (MTX) on Th1 and Th2 immune responses in mice. For this investigation, mice were immunized subcutaneously at the base of the tail with ovalbumin (OVA) emulsified with complete Freund's adjuvant (day 0). Varying doses of MTX were orally administered daily from days 0 to 20. On day 21, anti-OVA IgG2a and interferon-gamma (IFN gamma) as indicators of Th1 responses and anti-OVA IgG1 and interleukin-10 (IL 10) as those of Th2 responses were measured. The results showed that treatment with MTX was followed by decreases in OVA-specific IgG and proliferation of spleen cells to the antigen. The anti-rheumatic drug inhibited both anti-OVA IgG2a and IgG1 production, although the inhibitory effect of MTX on the antigen specific IgG2a production appeared to be greater than that on IgG1 production. IFN-gamma, but not IL-10, secretion was markedly downregulated by MTX. Administration of MTX resulted in suppression of antigen (OVA)-induced arthritis in mice. The suppression of the joint inflammation by MTX was associated with inhibition of OVA-specific proliferative responses of spleen cells, anti-OVA IgG, IgG2a and IgG1 production, and IFN-gamma and IL-10 secretion, although more pronounced decreases in IgG2a and IFN-gamma were observed compared with those in IgG1 and IL-10 in MTX-treated mice. These results indicate that MTX appears to suppress Th1 and, to a lesser extent, Th2 immune responses and its anti-arthritic effect on human rheumatoid arthritis might be at least in part explained by down regulation of Th1 responses involved in the disease. PMID- 14738594 TI - Study of inhibition of CYP2A6 by some drugs derived from quinoline. AB - CYP2A6 metabolizes coumarin to 7-hydroxycoumarin showing fluorescence, as measured by fluorometry. Firstly, we measured the inhibition of coumarin 7 hydroxylase of cDNA-expressed human CYP2A6 and in bovine liver microsomes, by quinoline and fluoroquinolines (FQ). Quinoline, 5-FQ, 6-FQ and 8-FQ inhibited activity but 3-FQ showed little inhibition. This suggests that the position 3 of quinoline is a recognition site for CYP2A6. We found similar patterns of coumarin 7-hydroxylase activity with human pooled liver microsomes. The level of CYP2A6 in human and bovine microsomes is the same as that detected by immunological titration with monoclonal antibody against CYP2A6. Secondly, we studied the inhibition of CYP2A6 with clinically used drugs of quinoline compounds, such as norfloxacin as an antibacterial agent, quinidine as an antiarrhythmic agent, quinine and chloroquine as antimalaria agents and rebamipide as an anti-ulcer agent. IC50 values (concentration producing 50% inhibition in activity) of norfloxacin, rebamipide and chloroquine at mM concentrations showed them to possess almost no inhibitory activity or influence on drug interaction. Meanwhile, the IC50 value of quinidine was 1.12 mM. The IC50 value of quinine was 160 microM with weak inhibition, suggesting that quinine, at a high dose, influences the metabolism of substrates for CYP2A6 by drug-drug interaction. These results also show that CYP2A6 discriminates the structure difference between the diastereoisomers quinidine and quinine. PMID- 14738595 TI - Effects of microcrystalline plant sterol suspension and a powdered plant sterol supplement on hypercholesterolemia in genetically obese Zucker rats. AB - Because dietary fat appears to be an effective vehicle for dispensing plant sterols into the diet, a special plant-sterol-containing ingredient has recently been developed. This ingredient is a plant sterol suspension in oil in which the sterols are in microcrystalline form. The objective of the present study was to analyse the cholesterol-lowering effects and safety of two different plant sterol preparations, an orally administered microcrystalline plant sterol suspension (MPS) in rapeseed oil and a powdered plant sterol supplement, in obese Zucker rats. Dietary plant sterol supplements (0.5%, w/w) were given concurrently with a high cholesterol diet (HCD, 1% cholesterol and 18% fat, w/w). No significant changes in serum triglyceride, blood glucose, serum glutamate oxaloacetic transaminase and glutamic pyruvic transaminase values or body and liver weights were observed. The powdered plant sterol supplement lowered the serum cholesterol by 25% (P < 0.05) and the MPS diet by 35% (P < 0.001) compared with HCD by the end of the 12-week experiment. Interestingly, the plant sterol supplements also produced a marked reduction in serum ubiquinone levels, suggesting a possible effect on isoprene synthesis. Unlike the powdered plant sterol, both MPS and plain rapeseed oil decreased the serum baseline diene conjugation values, suggesting that they protect against oxidative stress-induced lipid peroxidation in rats. This lipid peroxidation diminishing effect is probably due to some antioxidative components in rapeseed oil. These findings indicate that an unesterified plant sterol, such as the microcrystalline suspension in oil, effectively prevents cholesterol absorption in obese Zucker rats. PMID- 14738596 TI - In-vitro and in-vivo antioxidant activity of different extracts of the leaves of Clerodendron colebrookianum Walp in the rat. AB - The in-vitro antioxidant activities of different concentrations of the water, alcoholic, petroleum ether and ethyl acetate extracts of the dried leaves of Clerodendron colebrookianum Walp, and in-vivo antioxidant activity of the water extract was studied in experimental rat models. The results obtained from in vitro lipid peroxidation induced by FeSO4-ascorbate in rat liver homogenate showed a significant inhibition of lipid peroxidation by different extracts of C. colebrookianum leaf. Water extracts at concentrations (w/v) of 1:30, 1:50, 1:200 and 1:1000 showed the strongest inhibitory activity over the other organic extracts, suggesting maximum antioxidant effect. Chronic feeding of the water extract to Wistar albino rats (both sexes, 150-200 g) in 1 or 2 g kg-1/day doses for 14 days significantly increased the ferric reducing ability of plasma by 19% and 40% on the seventh day, and by 45% and 57% on the fourteenth day of treatment, respectively. Thiobarbituric acid reactive substances (TBARS), as a marker of lipid peroxidation, and some cellular antioxidants (superoxide dismutase, catalase and reduced glutathione) were estimated in heart, liver and kidney. There was a significant reduction in hepatic and renal TBARS with both the doses, without any change in myocardial TBARS. There was no change in the level of antioxidants in heart, liver and kidney, except for the hepatic superoxide dismutase. The findings of this study showed that the leaf extract of C. colebrookianum increased the antioxidant capacity of blood and had an inhibitory effect on the basal level of lipid peroxidation of liver and kidney. This lends scientific support to the therapeutic use of the plant leaves, as claimed by the tribal medicine of North-East India. PMID- 14738597 TI - Medicinal cannabis: is delta9-tetrahydrocannabinol necessary for all its effects? AB - Cannabis is under clinical investigation to assess its potential for medicinal use, but the question arises as to whether there is any advantage in using cannabis extracts compared with isolated Delta9-trans-tetrahydrocannabinol (Delta9THC), the major psychoactive component. We have compared the effect of a standardized cannabis extract (SCE) with pure Delta9THC, at matched concentrations of Delta9THC, and also with a Delta9THC-free extract (Delta9THC free SCE), using two cannabinoid-sensitive models, a mouse model of multiple sclerosis (MS), and an in-vitro rat brain slice model of epilepsy. Whilst SCE inhibited spasticity in the mouse model of MS to a comparable level, it caused a more rapid onset of muscle relaxation, and a reduction in the time to maximum effect compared with Delta9THC alone. The Delta9THC-free extract or cannabidiol (CBD) caused no inhibition of spasticity. However, in the in-vitro epilepsy model, in which sustained epileptiform seizures were induced by the muscarinic receptor agonist oxotremorine-M in immature rat piriform cortical brain slices, SCE was a more potent and again more rapidly-acting anticonvulsant than isolated Delta9THC, but in this model, the Delta9THC-free extract also exhibited anticonvulsant activity. Cannabidiol did not inhibit seizures, nor did it modulate the activity of Delta9THC in this model. Therefore, as far as some actions of cannabis were concerned (e.g. antispasticity), Delta9THC was the active constituent, which might be modified by the presence of other components. However, for other effects (e.g. anticonvulsant properties) Delta9THC, although active, might not be necessary for the observed effect. Above all, these results demonstrated that not all of the therapeutic actions of cannabis herb might be due to the Delta9THC content. PMID- 14738598 TI - In-vitro and in-vivo anti-inflammatory effect of oxyresveratrol from Morus alba L. AB - The antioxidative effects of mulberroside A and oxyresveratrol obtained from Mori Cortex were examined. Mulberroside A and oxyresveratrol showed an inhibitory effect against FeSO4/H2O2-induced lipid peroxidation in rat microsomes and a scavenging effect on 1,1-diphenyl-2-picrylhydrazyl radical. The anti-inflammatory effects of mulberroside A and oxyresveratrol using the carrageenin-induced model of inflammation were investigated in rats. Mulberroside A and oxyresveratrol significantly reduced paw edema. To investigate the mechanism of the anti inflammatory action of these compounds, we examined the effects of oxyresveratrol on lipopolysaccharide (LPS)-induced responses in murine macrophage cell line RAW 264.7. Exposure of LPS-stimulated cells to oxyresveratrol inhibited nitrite accumulation in the culture medium. Oxyresveratrol also inhibited the LPS stimulated increase of inducible nitric oxide synthase (iNOS) expression in a concentration-dependent manner; however, it had little effect on iNOS enzyme activity, suggesting that the inhibitory activity of oxyresveratrol is mainly due to the inhibition of iNOS expression rather than iNOS enzyme activity. Oxyresveratrol significantly inhibited LPS-evoked nuclear translocation of NF kappaB and cyclooxygenase-2 (COX-2) activity in RAW 264.7 cells. The results suggest that the anti-inflammatory properties of oxyresveratrol might be correlated with inhibition of the iNOS expression through down-regulation of NF kappaB binding activity and significant inhibition of COX-2 activity. PMID- 14738599 TI - Pharmacokinetics of thalidomide in patients with impaired renal function and while on and off dialysis. AB - There is a renewed interest in thalidomide for use in malignancies and systemic inflammatory diseases. Reduced renal function is not uncommon among patients with these disease states but the pharmacokinetics has not been fully investigated. The aim of this study was to investigate the pharmacokinetics of thalidomide in haemodialysis patients while on and off dialysis and in myeloma patients with varying degrees of renal function. Two studies were performed. To establish the pharmacokinetics of thalidomide in patients with mild to moderate renal failure, blood samples were taken over 12 weeks from 40 patients with multiple myeloma. A second study was performed in six patients with end-stage renal disease both on a non-dialysis day and before and during a haemodialysis session. Thalidomide concentration was determined by HPLC. A one-compartment open model with first order absorption and elimination was used to fit total thalidomide concentration to population pharmacokinetics and statistical models using the NONMEM program. Clearance and volumes were slightly below 10 L h-1 and 1 L kg-1, respectively, in both patient groups. The inter- and intra-patient variability was low. Clearance was doubled during dialysis. There was no correlation between thalidomide clearance and renal function. In conclusion, the pharmacokinetics of thalidomide in patients with renal failure are very similar to values reported by others for patients with normal renal function. Although clearance during dialysis is doubled, thalidomide dose need not be changed for patients with decreased kidney function. There is also no need for a supplementary dose due to haemodialysis. PMID- 14738601 TI - Management of lateral sinus thrombosis: update and literature review. AB - The management of four cases of lateral sinus thrombosis (LST) over a four-year period at the Royal Darwin Hospital is presented in this retrospective review. The patients were aboriginal and presented with otalgia, otorrhoea and sepsis. Two cases had an associated complication of an otitic hydrocephalus and a subperiosteal abscess. Cholesteatoma was found in three cases. Computed tomography (CT) scan confirmed the LST in three cases. Three patients were anaemic and thrombocytopenic. All patients had positive blood cultures. The organisms were predominantly mixed anaerobes and Bacteroides species. Three patients were managed surgically as a two-stage procedure. One patient was managed as a single-stage procedure with a modified radical mastoidectomy. Therapeutic anticoagulation was utilized in one case. There were no deaths. The prognosis of LST is good if treatment is instituted early with broad-spectrum intravenous antibiotics and surgery. The role of clot removal at surgery and the use of anticoagulation are discussed in this paper. PMID- 14738602 TI - Allergen immunotherapy in ENT: historical perspective. AB - The origins of immunology and allergy are founded upon the early 19th century microbiological studies of Jenner and Pasteur. It was discovered that the immune system could cause harm. The subspecialty of allergy began with the coining of the term by Von Pirquet in 1906 to describe disorders resulting from hyper reaction to normally innocuous environmental agents. Understanding the scientific basis of the immune system and allergy allowed Noon and Freeman, and later Cooke, to develop allergen immunotherapy. Initially the technique was crude, but with the subsequent key discovery of IgE, more accurate methods of diagnosis (such as the radioallergosorbent test (RAST)) and treatment ensued. The efficacy of specific immunotherapy has been demonstrated by many double-blind trials culminating in the WHO position paper. DNA recombinant technology has provided detailed molecular understanding of allergic disorders, which has resulted in several novel methods of immunotherapy that are potentially safer and more effective. Use of recombinant allergens, T-cell peptides, DNA vaccination with CpG motifs or plasmid vectors and anti-IgE strategies with monoclonal antibodies are showing promise. PMID- 14738603 TI - Infection, allergy and the hygiene hypothesis: historical perspective. AB - The 'hygiene hypothesis' was popularized in the late 1980s to explain the high prevalence of atopic disorders in the developed countries. It links atopic disorders and the lack of early life infections. An association between the two is not novel and dates back to the beginnings of allergy, immunology and microbiology. Allergy and infection have always been closely related and the study of one has often provided new insights into the pathobiology of the other. Early research into bacterial infections led to the discovery of the human immune system and the concept of allergy. An important relationship exists between parasite infections and the development of atopic disorders. This review traces the long and intimate historical relationship between infection and allergy. PMID- 14738604 TI - Mode of parotid involvement in external auditory canal carcinoma. AB - Until now, little was known about the mode of parotid involvement in external auditory canal (EAC) carcinoma. The incidence of parotid node metastasis and direct parotid invasion was examined in patients with EAC carcinoma. The study comprised 11 patients with squamous cell carcinomas (SCC) and 10 patients with adenoid cystic carcinomas (ACC). A retrospective review of the surgical specimens was undertaken with specific reference to parotid node metastasis and parotid invasion. Parotid node metastasis was noted only in two cases of advanced staged SCC, whereas none of the ACC patients showed parotid node metastasis. Direct parotid invasion occurred only in advanced staged SCC, however, it did occur in early stage ACC. Our data indicated that elective parotidectomy for control of occult parotid node metastasis is necessary only in advanced SCC carcinoma, whereas parotid management to secure adequate safety margins is mandatory for advanced SCC and all cases of ACC. PMID- 14738605 TI - Long-term socio-economic impact of vestibular schwannoma for patients under observation and after surgery. AB - This study describes and compares the long-term socio-economic impact for patients diagnosed with a vestibular schwannoma and either operated on or observed. A consecutive sample of patients diagnosed with vestibular schwannoma in Denmark and either operated on (748 patients) or observed by the wait-and-re scan policy (272 patients) during the period 1976-2000 were studied retrospectively. The consequences of operation/diagnosis (and observation) on vocational status, ability to handle daily chores and some psycho-social aspects were studied by means of a prospective postal questionnaire. Ninety-six per cent of the operated and 83 per cent of the observed patients answered the questionnaire. Overall, 34 per cent of operated patients resumed their daily activities within one to two months, and 76 per cent within four to six months. Patients operated on for a large tumour resumed their daily activities later than patients with a small tumour. Regardless of tumour size, employment was unchanged for the majority of observed and operated patients. The vocational consequences were significantly worse for operated patients with a large tumour, than for observed patients. However, no difference existed between the observed group and operated patients with a tumour below 20 mm in size. A change in vocational status was most frequent for assisting spouses, unskilled manual workers and the self-employed. The majority of both observed and operated patients experienced no change in their ability to handle daily chores. The changed ability of operated patients was worse than that of observed patients. Among various changes in their psycho-social well-being, decrease in social ability was the most frequent complaint in both groups, followed by increased fatigue, decreased concentration, increased irritability, depression and headache, decreased intellect and libido. Regardless of tumour size, the change in social ability, concentration and fatigue was worse for operated patients. Concerning headache, patients operated on for a large tumour were better off than observed patients and patients operated on for a small tumour. There was no difference between the operated and observed groups concerning irritability, intellect and libido. Deterioration of vocational status, ability to handle daily chores and several aspects of psycho social well-being are reported both by patients operated on and observed for vestibular schwannoma. However, the negative changes were more frequent among the operated patients, although the differences were surprisingly modest, especially when comparing observed patients with patients operated on for a small tumour. PMID- 14738606 TI - Technique to prevent post-operative CSF leak in the translabyrinthine excision of vestibular schwannoma. AB - Cerebrospinal fluid (CSF) leak following VS removal is a challenging complication. With experience, it can be seen that anatomical factors such as the extent of pneumatization of the petrous temporal bone and posterior wall of the internal auditory canal play an important role in contributing to CSF leak. Nevertheless, the risk of post-operative CSF leak remains a major concern for both the surgeon and patient. This paper describes a technique, which we have used since 1994, and which has helped us to achieve the lowest reported rate of post-operative CSF leak in translabyrinthine excision of vestibular schwannoma in the world literature. PMID- 14738607 TI - Incidental magnetic resonance image sinus abnormalities in asymptomatic Australian children. AB - Plain X-rays, computed tomography (CT) and magnetic resonance imaging (MRI) scans performed for non-ENT reasons often reveal incidental sinus mucosal changes. These changes need to be correlated clinically before diagnosing rhinosinusitis. This study examined the prevalence of such changes in MRI scans in children up to age 16. Scans were scored using an adapted Lund-Mackay classification and were positive when one or more sinuses showed abnormalities. Randomly selected scans in the retrospective arm revealed a prevalence of 20 of 62 (32.3 per cent). In the prospective arm 45 of 60 children were defined as truly asymptomatic, of which 14 scans (31 per cent) were positive. Other studies in adults and children using CT and MRI report a prevalence range of roughly 30 to 45 per cent. This variability may be attributed to differences of study design, definitions of population age, definitions of asymptomatic and definition of abnormal sinus. Other plausible factors to explain regional differences are climate and frequency of upper respiratory tract infections. PMID- 14738608 TI - Fine needle aspiration in chronic tonsillitis: reliable and valid diagnostic test. AB - Fine needle aspiration (FNA) of the tonsil as a diagnostic tool in evaluating the microflora in chronic tonsillitis has not been popularized. A prospective study of 30 patients with chronic tonsillitis undergoing tonsillectomy was undertaken. FNA of the tonsil core was done under local/general anaesthesia. The reliability of the culture by FNA of the tonsil core was then validated with the reference (gold) standard which is the dissected tonsil core. The sensitivity of FNA culture as compared to core culture was 100 per cent and 93 per cent under general and local anaesthesia respectively. The positive predictive value of FNA culture as compared to core culture was 92 per cent and 82 per cent for general and local anaesthesia respectively. These factors indicate that FNA of the tonsil core is reliable and valid. It can safely be performed as an out-patient procedure under local anaesthesia. This is reported for the first time. Identifying the bacterial organism within the infected tonsil for appropriate antibiotic therapy could revolutionize the management of chronic tonsillitis. PMID- 14738609 TI - Pre- and intra-operative staging of the neck in a developing world practice. AB - Most patients with squamous cancer of the head and neck treated at Groote Schuur Hospital are from a poor socio-economic background with a high prevalence of tuberculosis (TB), human immunodeficiency virus (HIV) and other infections that may cause cervical lymphadenopathy resulting in overstaging of the neck. A retrospective review of 186 patients requiring therapeutic and elective neck dissection was undertaken and the sensitivity and specificity of clinical and intra-operative staging of the neck determined. Results showed overall sensitivity of staging at 80.1 per cent. Specificity was 52.2 per cent. Staging of the N(1), N(2b) and N(2c) necks had positive predictive values of 53.2, 65.8 and 68.2 per cent respectively. Occult nodal metastases were present in 32 per cent elective neck dissections (END)s. Specificity of intra-operative staging of the N(0) neck was 33.3 per cent and sensitivity was 72.4 per cent. Conclusions were that our indications for elective neck dissection are appropriate. The high false positive rates for staging of the N(1), N(2b) and N(2c) necks, necessitate a change in management strategy. PMID- 14738610 TI - Prevalence of back and neck pain amongst ENT consultants: national survey. AB - Back and neck pain is an established occupational disease for many professions. No previous studies have shown the association of back and neck pain with ENT surgery. Our objective was to determine the prevalence of neck and back pain in ENT consultants in the United Kingdom and which sub-specialties were most at risk. A national survey from 325 ENT consultants in the UK revealed that 72 per cent had either back pain or neck pain or even both. Of those with pain 53 per cent attributed their symptoms directly to ENT surgery. Highest figures were received from otologists, related to microscope work and prolonged sitting. The importance of these findings has demonstrated the lack of awareness and adequate training to prevent long-term disability in ENT surgeons. PMID- 14738611 TI - Management of an infected preauricular sinus, using a lacrimal probe. AB - This paper describes a technique for managing a preauricular abscess that allows drainage of the pus, with little or no disturbance to the underlying sinus, making any subsequent surgery more straight forward. PMID- 14738612 TI - Emergency management of tracheo-brachiocephalic artery fistula. AB - The authors present a technique for emergency management of a tracheo brachiocephalic artery fistula. PMID- 14738613 TI - Chorda tympani neuroma masquerading as cholesteatoma. AB - Facial nerve neuromas occur throughout the course of the facial nerve and its branches, however lesions occurring on the chorda tympani branch are exceptionally rare. We present a case where the diagnosis was made intra operatively; the patient was pre-operatively thought to have had a cholesteatoma. Total resection is the treatment of choice for these cases. Early diagnosis, aided by high resolution computed tomography (CT) scanning, will facilitate complete excision without damage to the facial nerve itself or the ossicular chain. The slow growing nature of the neuroma is likely to allow compensatory mechanisms to occur without the patient experiencing dysgeusia. As with any rarity the diagnosis can only be made with a high index of suspicion. PMID- 14738614 TI - Aggressive papillary tumour of the nasopharynx followed by an aggressive papillary tumour of the middle ear. A multiple site tumour? AB - We report the case of a 72-year-old male presenting with a papillary adenocarcinoma of the middle ear. He had had a similar tumour excised from the ipsilateral nasopharynx seven years previously with no evidence of local recurrence. We conclude that this middle ear lesion possibly represents a second tumour. No record of such a case has been reported previously in the literature. We discuss presentation and management and highlight the need for close follow-up of these patients. PMID- 14738615 TI - Septal perforation secondary to Mycobacterium kansasii infection. AB - We report the first recorded case of a septal perforation caused by Mycobacterium kansasii. This atypical mycobacterium is finding increasing prevalence with the increasing incidence and longevity of human immunodeficiency (HIV) infections. Cases of chest infection, sinusitis, septic arthritis, osteomyelitis, pericarditis, brain abscess, cutaneous and oral lesions have all now been reported. This discovery represents a rare but important differential in the aetiology of septal perforation. PMID- 14738616 TI - Relapsing acute adult epiglottitis following hypophysectomy. AB - Adult epiglottitis is an uncommon disease that may become fatal because of sudden airway obstruction. Recurrent adult epiglottitis has been reported rarely in the literature. We present a case of relapsing epiglottitis in a patient with corticosteroid insufficiency secondary to pituitary surgery. It is the first case to highlight this infection occurring in a relapsing manner in a patient with pituitary-adrenocortical insufficiency on long-term steroid replacement therapy. PMID- 14738617 TI - Mucoepidermoid carcinoma of the larynx: report of three cases. AB - Mucoepidermoid carcinoma of the larynx is very rare, with only a few individual cases or studies in small patient groups reported in the literature. Treatment modalities for this type of carcinoma are not uniform; in addition, errors in the recognition and pathologic diagnosis are not uncommon. During the period 1991 2000, 771 cases of malignant laryngeal tumours were recorded and histologically verified at the University Department of Otorhinolaryngology and Cervicofacial Surgery, Zagreb University Hospital Centre, three of them were mucoepidermoid supraglottic carcinomas. The experience acquired in the treatment of these tumours is presented along with a review of the literature. PMID- 14738618 TI - Angiomyoma in the submandibular gland: a rare location for a ubiquitous tumour. AB - Angiomyoma is a common soft tissue tumour of the head and neck that sometimes presents to the otolaryngologist; however, it seldom occurs in the major salivary glands. We present a case of angiomyoma arising in the submandibular gland, a tumour not described previously in the English literature. PMID- 14738619 TI - Osteoradionecrosis of the hyoid presenting as a cause of intractable neck pain following radiotherapy and the role of magnetic resonance image scanning to aid diagnosis. AB - Osteoradionecrosis of the hyoid has been reported rarely in the worldwide literature. We present the case of a 56-year-old gentleman who presented with intractable neck pain, following surgery and radiotherapy for a T(2)N(2c)M(0) tongue base carcinoma, to highlight the need to consider osteoradionecrosis of the hyoid rather than recurrence of the carcinoma as the cause of such symptoms. The previously unreported appearance of osteoradionecrosis of the hyoid on a magnetic resonance image (MRI) scan and the use of this investigation to aid diagnosis is discussed. PMID- 14738620 TI - Tinnitus due to clarithromycin. PMID- 14738621 TI - Analytical and clinical evaluation of CYFRA 21-1 by electrochemiluminescent immunoassay in head and neck squamous cell carcinoma. PMID- 14738622 TI - Cholesterol granuloma of the frontal sinus. PMID- 14738623 TI - Naseptin nasal cream 'contains peanut oil'. PMID- 14738625 TI - Marked hydroureter misdiagnosed as a hydrosalpinx by ultrasound. PMID- 14738627 TI - What your statistician never told you about P-values. AB - We provide a non-technical overview of what P-values are and what they are not. To determine how P-values ought to be used, reported, and interpreted, we must first clarify the often-overlooked differences between, and proper usages of, significance testing and hypothesis testing. Several clinical examples are given to illustrate these differences, and failure to distinguish between them is seen to be problematic. Common misinterpretations of P-values are explained. Confidence intervals provide essential information where P-values are deficient in doing so and they therefore play an essential role in reporting and interpreting study results. PMID- 14738628 TI - Factors affecting capacitive current diversion with a uterine resectoscope: an in vitro study. AB - STUDY OBJECTIVE: To evaluate electrosurgical waveform, generator type, and electrode integrity as variables in capacitive induction of current on the external sheath of a resectoscope in open-circuit conditions. DESIGN: In vitro, laboratory, comparative study (Canadian Task Force classification CII-1). SETTING: Surgical laboratory. INSTRUMENTATION: Three ValleyLab radiofrequency (RF) electrosurgical generators (ESU), Force-2, Force-4, and Force-F/X; a resectoscope (Storz 50 series); and rollerball electrodes, both intact and with two types of standard insulation defects (lateral and circumferential) placed in two locations: distally, beyond the end of the telescope, and proximally, beside the distal aspect of the telescope. A Dynatek ESU analyzer was used to record current and wattage on electrodes and the external sheath. MEASUREMENTS AND MAIN RESULTS: With intact insulation, current was not disproportionately induced on the external sheath of the resectoscope regardless of ESU, power, or waveform. Proximally located electrode insulation defects allowed induction of most of the generator's output to the external sheath when high-voltage modulated outputs were used, and the risk varied somewhat with the ESU. There was no such induction at any power setting or with any insulation defect when low-voltage (cutting) outputs were tested. CONCLUSION: In the presence of proximal electrode defects, high-voltage currents may contribute to thermal injury to the lower genital tract during RF resectoscopic surgery. PMID- 14738629 TI - Laparoscopic excision of very large myomas. AB - STUDY OBJECTIVE: To evaluate the feasibility, complications, and conversion rate of laparoscopic excision of very large myomas. DESIGN: Prospective study (Canadian Task Force classification II-2). SETTING: Private endoscopy center. PATIENTS: Fifty-one women with at least one myoma larger than 9 cm. INTERVENTION: Laparoscopic myomectomy. MEASUREMENTS AND RESULTS: We removed 78 myomas laparoscopically in these 51 patients. Three patients had two myomas larger than 9 cm, three had two myomas between 5 and 9 cm (in addition to 1 > 9 cm), and one had three myomas between 5 and 9 cm (in addition to 1 > 9 cm). Mean number of myomas removed/patient was 1.53 +/- 1.17 (range 1-6); 12 women (23.5%) had multiple myomectomy. The largest myoma removed was 21 cm. Mean myoma weight was 698.47 +/- 569.13 g (range 210-3400 g). Mean operating time was 136.67 +/- 38.28 minutes (range 80-270 min). Mean blood loss was 322.16 +/- 328.2 ml (range 100 2000 ml). One patient developed a broad ligament hematoma, two developed postoperative fever, and one underwent open subtotal hysterectomy 9 hours after surgery for dilutional coagulopathy. CONCLUSION: Myomectomy by laparoscopy is a safe alternative to laparotomy for very large myomas. PMID- 14738630 TI - Risk factors for conversion to laparotomy during gynecologic laparoscopy. AB - STUDY OBJECTIVE: To identify risk factors and short-term outcomes associated with conversion from laparoscopic surgery to laparotomy. DESIGN: Case control study (Canadian Task Force classification II-2). SETTING: Large urban, academic medical center. SUBJECTS: All 2530 gynecologic laparoscopies, including those converted to laparotomy. INTERVENTION: Laparoscopic surgery. MEASUREMENTS AND MAIN RESULTS: Statistical analysis was performed to compare risk factors in converted versus non-converted cases. Multiple logistic regression analysis was performed using variables found to be significant on univariate analysis. Of 2530 laparoscopies identified, 159 (6.3%) were converted to laparotomy; 323 were selected by random number assignment from the remaining cases for the control group. The following factors were significantly associated with unintended laparotomy (OR; 95% CI): surgeon inexperience (2.43; 1.38, 4.17), level of laparoscopic complexity (3.19; 1.20, 5.10), body mass index greater than 30 kg/m(2) (2.45; 1.40, 4.41), suspicion of malignancy (17.45; 7.32, 43.57), history of laparotomy (1.34; 1.02, 1.78), presence of adhesions (2.30; 1.37, 3.76), and intraoperative technical difficulty (17.86; 7.32, 43.57). When stratified by level of complexity, experience in laparoscopy does not appear to confer protection during level 1 laparoscopic procedures (0.42; 0.12, 1.37) but does significantly reduce the frequency of conversion during level 2 procedures (0.39; 0.72, 0.93). Age, parity, bowel injury, vascular injury, and history of pelvic inflammatory disease and endometriosis did not increase the risk of conversion. Compared with controls, patients who were converted experienced significantly greater blood loss (p < 0.001), longer operating room time (p < 0.001), and longer hospital stay (p < 0.001). CONCLUSION: All women undergoing laparoscopy should be counseled that unintended laparotomy is a known risk and has additional morbidity over laparoscopy alone. Less-experienced surgeons attempting complicated procedures significantly increase the risk of conversion. PMID- 14738631 TI - Comparison of laparoscopy and laparotomy in managing hemodynamically stable patients with ruptured corpus luteum with hemoperitoneum. AB - STUDY OBJECTIVE: To compare differences between laparoscopy and laparotomy in managing women ruptured corpus luteum with hemoperitoneum. DESIGN: Three-year, prospective, nonrandomized study (Canadian Task Force classification II-2). SETTING: University-affiliated regional hospital. PATIENTS: Sixty hemodynamically stable women. INTERVENTION: Laparoscopic surgery (30 women) and laparotomy (30). MEASUREMENTS AND MAIN RESULTS: Laparoscopic surgery had significant advantages over laparotomy, including shorter hospital stay (55.33 +/- 7.67 vs 97.77 +/- 14.45 hrs, p < 0.001) without increased adverse events. Laparoscopic surgery also showed trends of shorter operating time, improved wound care, and less postoperative pain. CONCLUSION: Laparoscopy surgery for diagnosis and treatment of women with ruptured hemorrhagic corpus luteum appears superior to laparotomy. We suggest that surgeons try laparoscopy first as a diagnostic and probably therapeutic procedure. PMID- 14738632 TI - Laparoscopic appendectomy in an outpatient setting. AB - STUDY OBJECTIVE: To determine the ability to perform laparoscopic appendectomy on an outpatient basis. DESIGN: Prospective study (Canadian Task Force classification II-2). SETTING: University ambulatory surgery center. PATIENTS: Forty-two consecutive patients. INTERVENTION: Laparoscopic appendectomy. MEASUREMENTS AND MAIN RESULTS: After performing five laparoscopic appendectomies, three women required hospitalization. Thus a new multimodal protocol was implemented and admission and recovery times were monitored. After the new protocol was implemented, only 1 of 37 patients required hospitalization. Discharge times were similar to those for women undergoing gynecologic laparoscopy without appendectomy. CONCLUSION: With attention to surgical and anesthetic protocols, appendectomy can be performed at the time of gynecologic laparoscopy in an ambulatory setting without a significant delay in discharge. PMID- 14738633 TI - Outpatient total laparoscopic hysterectomy. AB - STUDY OBJECTIVE: To assess the safety of and patient satisfaction with total laparoscopic hysterectomy as an outpatient procedure. DESIGN: Retrospective case study (Canadian Task Force classification III). SETTING: Tertiary care hospital. PATIENTS: Sixty-six consecutive women. INTERVENTION: Outpatient total laparoscopic hysterectomy. MEASUREMENTS AND MAIN RESULTS: Of 66 procedures completed, 6 patients (7.5%) required overnight hospitalization, with 4 of them discharged the next day. Seven (11%) minor postoperative complications occurred. One patient required hospitalization to receive intravenous antibiotics and one for drainage of a cuff hematoma. There were three (4.5%) minor intraoperative complications. One woman developed Clostridium difficile diarrhea as well as deep vein thrombosis and pulmonary embolus. Sixty-three women (95%) were satisfied with the procedure and would recommend it to others. CONCLUSION: Outpatient total laparoscopic hysterectomy is well tolerated, safe, and cost effective. PMID- 14738634 TI - A retrospective study evaluating the impact of formal laparoscopic training on patient outcomes in a residency program. AB - STUDY OBJECTIVE: To evaluate the impact of a formal laparoscopic training program on patient outcomes in an obstetrics and gynecology residency. DESIGN: Retrospective cohort study (Canadian Task Force classification II-1). SETTING: University-affiliated public hospital. INTERVENTION: Comparison of nonlaparoscopy trained group (A) with a formally trained group (B). SUBJECTS: Three hundred sixty-eight women (group A) and 154 women (group B) undergoing operative laparoscopy. MEASUREMENTS AND MAIN RESULTS: Beginning October 1, 1999, residents, regardless of postgraduate year status, participated in six 4-hour sessions/year in a committed laparoscopic training program consisting of didactics, bench exercises (designed to mimic laparoscopic gynecologic technical skills), instrumentation instruction, animate tissue model surgery, and supervised gynecologic operating experience. Two certified gynecology laparoscopists facilitated each training session, and gynecologists with various laparoscopic skill levels supervised patient surgeries. Operating room time, blood loss, hospital stay, and conversion to laparotomy were less for group B than for group A, but the groups did not differ in complication rates. Adhesions and bleeding were the main reasons for conversion to laparotomy. CONCLUSION: Formal laparoscopic training of gynecologic residents improved patient outcomes. PMID- 14738635 TI - Lateral distribution of ovarian dermoid cyst. AB - STUDY OBJECTIVE: To determine the distribution of ovarian dermoid cysts in a large series of women. DESIGN: Observational study (Canadian Task Force classification II-3). SETTING: University teaching hospitals. PATIENTS: Three hundred thirty-six women. INTERVENTION: Evaluation of operative reports. MEASUREMENTS AND MAIN RESULTS: Dermoid cysts had a mean diameter of 5.6 cm. They were present in the right ovary (48.8 %), left ovary (37.8%), and bilaterally (13.4%). Unilateral cysts were more frequent in the right ovary (56.4%) than in the left (p = 0.002, odds ratio 1.7, confidence interval 1.2-2.3). This was significantly different from the expected 50% (4.45, chi2). A similar predisposition was found when we included women with bilateral cysts (p < 0.01, OR 1.5, CI 1.1-1.9). Patient age and size of cyst did not differ between those with left and right lesions. CONCLUSION: In this large cohort of women, ovarian dermoid cysts had a predisposition to a right lateral location. PMID- 14738636 TI - Quality of life should be considered the primary outcome for measuring success of endometrial ablation. AB - STUDY OBJECTIVE: To compare clinical and quality of life outcomes by validated instruments after endometrial ablation by four methods, and determine the primary outcome that most reflects procedure success. DESIGN: Descriptive cohort study (Canadian Task Force classification II-2). SETTING: Minimal-access gynecologic surgery unit in northeast England. PATIENTS: One hundred thirty-nine women. INTERVENTION: Endometrial ablation by one of four methods: Cavaterm, endometrial laser interstitial thermotherapy (ELITT), endometrial laser ablation (ELA), or NovaSure impedance-controlled system. MEASUREMENTS AND MAIN RESULTS: Clinical outcomes were assessed according to menstrual outcome. Whereas amenorrhea rates were significantly different among the four techniques (chi2 = 9.42, p = 0.02), combined amenorrhea and hypomenorrhea rates were not significantly different (67%, 69%, 69%, 62%, chi2 = 0.38, NS) for Cavaterm, ELITT, ELA, and NovaSure, respectively. Quality of life, measured preoperatively by the EQ-5D index and EQ 5D vas, SF-12 physical and mental component scores, and sexual activity questionnaire pleasure, habit, and discomfort scores, were substantially improved 12 months after treatment. No significant differences were seen in quality of life scores between women after endometrial ablation and the general population. CONCLUSION: Quality of life for women with menorrhagia is improved to a normal level by treatment with either first- or second-generation endometrial ablation. PMID- 14738637 TI - Laparoscopic suburethral sling procedure. AB - STUDY OBJECTIVE: To describe the technique of laparoscopic suburethral sling and discuss patient satisfaction and subjective continence rates over 7 years. DESIGN: Retrospective analysis (Canadian Task Force classification III). SETTING: Private practice. PATIENTS: Sixty-three consecutive patients. INTERVENTION: Laparoscopic sling procedure using either autologous or donor fascia. MEASUREMENTS AND MAIN RESULTS: The space of Retzius is entered laparoscopically. Then from a vaginal approach, a small midline vertical incision is made below the urethra, and vaginal epithelium is dissected off underlying periurethral and paravesical fascia. Material for the sling is passed underneath the urethra and secured laparoscopically to Cooper's ligaments bilaterally. Over 7 years, 63 laparoscopic suburethral sling procedures were performed. Overall, patients reported 88.9% satisfaction rate and 81.0% subjective continence rate. CONCLUSION: The laparoscopic approach is an acceptable alternative to other suburethral sling procedures. PMID- 14738638 TI - Bilateral laparoscopic transperitoneal ligation of ovarian veins for treatment of pelvic congestion syndrome. AB - STUDY OBJECTIVE: To evaluate the efficacy and safety of bilateral laparoscopic transperitoneal ligation of the ovarian veins in women with symptomatic pelvic varices. DESIGN: Prospective pilot study performed in a small series (Canadian Task Force classification III). SETTING: Urban hospital in Turin and University hospital in Cagliari, Italy. PATIENTS: Twenty-three women. INTERVENTION: Bilateral laparoscopic transperitoneal ligation of the ovarian veins. The right ovarian vein was reached by incising posterior peritoneum below the mesentericoparietal fossa in all women. The left ovarian vein was reached by reflecting medially the left colon in 10 women and by incising the posterior peritoneum covering the aorta 2 cm below the inferior duodenal fold in 13. MEASUREMENTS AND MAIN RESULTS: After the plateau of the learning curve was reached, average operating time was about 60 minutes without complications. Complete remission of pain and absence of pelvic varicosities lasted for 12 months. CONCLUSION: The laparoscopic transperitoneal paraaortic approach to bilateral ligation of ovarian veins near their origin could be a new treatment option for pelvic congestion syndrome. PMID- 14738639 TI - Evaluation of HydroThermAblator and rollerball endometrial ablation for menorrhagia 3 Years after treatment. AB - STUDY OBJECTIVE: To compare the safety and efficacy of endometrial ablation using HydroThermAblator (HTA) and rollerball (RB) for treatment of menorrhagia. DESIGN: Prospective, randomized, multicenter study (Canadian Task Force classification I). SETTING: Nine private practice and university centers in the United States. PATIENTS: Two hundred seventy-six women. INTERVENTION: Hysteroscopic endometrial ablation with the HTA (187 women) and RB (89). MEASUREMENTS AND MAIN RESULTS: Bleeding was assessed by pictorial diaries for 12 months, with patient interviews at 24 and 36 month. Amenorrhea rates, reduction of bleeding to normal levels or less, and patient satisfaction were tracked for 36 months, with rates in the HTA group of 53%, 94%, and 98% and in the RB group of 46%, 91%, and 97%, respectively. CONCLUSION: Endometrial ablation with the HTA is a safe, effective, and durable treatment of menorrhagia in a broad patient population. It offers advantages over RB by reducing anesthesia requirements, reducing operating time, and eliminating risks of excessive fluid absorption, and is more easily learned. PMID- 14738640 TI - Insertion of mirena after endometrial resection in patients with adenomyosis. AB - STUDY OBJECTIVE: To evaluate the efficacy of Mirena, a levonorgestrel-releasing intrauterine device, after endometrial resection for treatment of menorrhagia caused by adenomyosis. DESIGN: Open, randomized, observational study (Canadian Task Force classification II-2). SETTING: Private hospital. PATIENTS: Ninety-five women. INTERVENTION: Endometrial resection, after which control patients received no further treatment and study patients had Mirena inserted immediately after the procedure. MEASUREMENTS AND MAIN RESULTS: The rate of amenorrhea after 1 year was significantly higher in the Mirena group. Nineteen percent of women in the control group had a second procedure to control bleeding compared with none in the Mirena group. CONCLUSION: Insertion of Mirena after endometrial resection is effective treatment for menorrhagia caused by adenomyosis and has very few adverse effects. PMID- 14738641 TI - Long-term follow-up of endometrial ablation by modified loop resection. AB - STUDY OBJECTIVE: To describe endometrial ablation by rollerbar-loop-rollerbar (RLR) technique and determine the procedure's long-term results. DESIGN: Prospective observational study (Canadian Task Force classification II-2). SETTING: Private gynecology practice. PATIENTS: One hundred seventeen women with menorrhagia not related to malignancy who were not interested in future fertility. INTERVENTION: After performing rollerbar ablation through a short large Graves speculum, loop resection was carried out down to and throughout superficial myometrium. Rollerbar redesiccation of the cornua and supracervical areas of the uterus completed the procedure. MEASUREMENTS AND MAIN RESULTS: Patients were contacted by telephone or mail 4 to 5 years after RLR and asked to complete a questionnaire to assess their bleeding status, level of satisfaction with the procedure, and whether or not they had undergone hysterectomy. Of 109 patients responding, 60 (55%) were amenorrheic or menopausal with no bleeding, 21 (19%) had spotting, and 17 (16%) had undergone hysterectomy. Eighty-four (79%) were very satisfied. CONCLUSION: RLR endometrial ablation was associated with a high rate of amenorrhea and substantial long-term patient satisfaction, but the rate of postablation hysterectomy remained significant. PMID- 14738642 TI - The HydroThermAblator system for management of menorrhagia in women with submucous myomas: 12- to 20-month follow-up. AB - STUDY OBJECTIVE: To assess the safety and efficacy of the HydroThermAblator (HTA) system to treat patients with menorrhagia and submucous myomas up to 4 cm in diameter. DESIGN: Retrospective, multicenter, cohort study (Canadian Task Force classification II-2). SETTING: Two medical centers from a large, prepaid, multispecialty health maintenance organization. PATIENTS: Twenty-two women. INTERVENTION: HTA endometrial ablation. MEASUREMENTS AND MAIN RESULTS: The HTA circulates free-flowing, heated (90 degrees C) saline in the uterine cavity for 10 minutes under direct hysteroscopic control. All procedures were performed in the office under local anesthesia after premedication with oral anxiolytic and analgesic agents. At least 1 year after treatment (average follow-up 15.4 mo, range 12-20 mo), women were contacted regarding their menstrual history. Success was defined as no additional medical or surgical intervention. Amenorrhea was defined as no bleeding, eumenorrhea as normal periods, and oligomenorrhea as less than a normal period. Twelve patients (54%) reported complete amenorrhea; 7 were premenopausal (41% amenorrhea rate) and 5 were postmenopausal (100% amenorrhea rate). Five (23%) reported oligomenorrhea and three (14%) eumenorrhea, and were very satisfied with the result. The overall success rate was 91%. Of two failures (9%), one woman had a repeat HTA 13 months later for menorrhagia. The other had a repeat HTA for persistent spotting and sustained a false passage and small perforation before the procedure began. She elected to have a vaginal hysterectomy 1 month later. Although the results are not statistically significant, patients were extremely satisfied with the outcomes, and no serious complications were related to the device or its use for this indication. CONCLUSION: HTA is probably safe and effective in treating women with menorrhagia and submucous myomas up to 4 cm in diameter. It offers advantages over hysterectomy and electrosurgical resection since it is an office-based procedure, and significant advantages over other global ablation technologies that, by their design, are unable to treat significantly distorted uterine cavities. PMID- 14738643 TI - Vessel sealing using a pulsed bipolar system and open forceps. AB - We evaluated the vessel-sealing ability of one application of a pulsed plasma kinetic (PK) electrosurgical device on skeletonized vessels up to 7 mm in diameter to withstand sustained pressure of 300 mm Hg using bench and animal models. Forty-six porcine specimens and 66 abattoir vessels (112 total) were pressure tested after sealing with a pulsed PK system. Of these, 108 (96%) met the criterion of 300 mm Hg pressure held for 10 seconds without leaking. The PK system open forceps effectively closes vessels and maintains closure at a pressure level sufficient for clinical use. The device is more effective than clips, sutures, and staples. PMID- 14738644 TI - Measurement of trocar insertion force using a piezoelectric transducer. AB - We attempted to establish a model to measure the force required for trocar insertion at laparoscopy. A 3-cm, circular transducer was constructed from piezoresistive material that changes its impedance as force is exerted on its surface. The transducer is connected by an interface box to a personal computer to record surface contact pressure digitally (pressure = force/area) profile continuously during trocar insertion. Each subject had three trocars inserted: a 10-mm trocar at the umbilicus after creation of pneumoperitoneum, and 5-mm trocars at corresponding sites on the left and right sides of the lower abdomen. All insertions were performed by the same operator using reusable trocar with a conical tip. Each subject acted as her own control. Recordings were successfully obtained from eight women. There was no instance of transducer failure. The mean (SE) peak contact surface pressure for the 10-mm and 5-mm left and right trocars were 5.3 (0.32), 6.4 (0.51), and 6.81 (0.27) pounds/square inch, respectively. Placement of the 10-mm trocar required less insertion force than placement of the 5-mm trocars. There was a strong negative correlation (r = -0.97, p < 0.001) between body weight and peak insertion force for the 10-mm trocar. PMID- 14738645 TI - Laparoscopic management of a translocated intrauterine device perforating the bowel. AB - A woman underwent operative laparoscopy to manage a dislocated intrauterine device that perforated her bowel. The surgical procedure was uncomplicated, and postoperative recovery was uneventful. PMID- 14738646 TI - Endometrial intraepithelial neoplasia diagnosed at endometrial resection. AB - The literature has no reported cases of isolated endometrial intraepithelial neoplasia found at endometrial resection. If endometrial cancer is occult it might not be detected at diagnostic hysteroscopy or during resection, especially if destructive techniques are used. A 51-year-old woman had history of menorrhagia, with diagnostic hysteroscopy showing benign functional endometrium and diagnosed as dysfunctional uterine bleeding. Endometrial resection was performed and the pathologic examination in one stripe of resected tissue found focal, isolated endometrial intraepithelial neoplasia. This case reinforces the importance of pathologic tissue examination after endometrial ablation. Care must be taken when performing second-generation nonhysteroscopic ablation techniques, as even with direct visualization a premalignant lesion can be missed. PMID- 14738647 TI - Laparoscopic approach to an uncommon adnexal neoplasm associated with infertility: serous cystadenofibroma of the fallopian tube. AB - A rare case of serous cystadenofibroma of the fallopian tube was discovered during evaluation for in vitro fertilization-embryo transfer. Bilateral tubal occlusion was noted on hysterosalpingogram, and a right adnexal cyst, initially thought to be of ovarian origin, was identified by office transvaginal sonography. Laparoscopy revealed a 5.5-cm, fluid-filled mass involving the distal aspect of the right fallopian tube. Both ovaries and uterine exterior appeared grossly normal. The cyst was decompressed and removed intact without incident through a 5-mm laparoscopic cannula. The mass showed histologic features consistent with benign serous cystadenofibroma. The patient had an uncomplicated postoperative convalescence and continued to do well 3 months after surgery. PMID- 14738648 TI - The utility of urodynamic testing. AB - The utility of urodynamic testing has been questioned for decades. However, this diagnostic tool has been widely adopted in the evaluation of patients who suffer from urinary incontinence despite the associated costs to society. A review of the literature and the consensus of the authors with regard to the indications of urodynamic testing, its application as a diagnostic and prognostic tool, and its effect on surgical management and outcome are presented here. PMID- 14738649 TI - The time course of myometrial ischemia and reperfusion after laparoscopic uterine artery occlusion--theoretical implications. PMID- 14738650 TI - How frequently should cervical screening be conducted - important new evidence. PMID- 14738651 TI - Is there a case for hepatitis C infection screening in the antenatal period? AB - Based on research evidence the UK National Screening Committee recently recommended that routine antenatal screening for hepatitis C virus (HCV) infection should not be introduced into the UK antenatal screening programme. In this paper we review the evidence on which this decision was based, addressing the criteria that need to be considered before the introduction of a new screening programme. PMID- 14738652 TI - A pilot study of 'fast track' antenatal screening for haemoglobinopathies. AB - The NHS Plan launched in 2000 advocated a linked antenatal and neonatal screening programme for haemoglobinopathies. Currently screening practices vary widely across the UK and patient sampling is generally performed in a hospital setting. The process is flawed and frequently fails to provide accurate and timely information. In this study we demonstrate that organisational changes can improve the efficiency and quality of screening. The primary care screening process described here has increased partner testing rates and allowed early identification of at risk couples. PMID- 14738653 TI - Testing for toxoplasmosis in pregnancy: a study of the routines in primary antenatal care. AB - OBJECTIVES: The question of benefit of screening for antibodies against Toxoplasma gondii in pregnancy to prevent impairment in the infant has not been settled. Despite this, it is believed that such testing is common. The aim of this study was to describe the test routines and to identify factors associated with testing in primary antenatal care. SETTING: Ulleval University Hospital, Oslo, Norway. METHODS: All women who gave birth at Ulleval University Hospital, Oslo, Norway, in May 2000 were included. The number of antibody tests during pregnancy was obtained from the patient registry at the department of microbiology at the same hospital. RESULTS: 81% (361/446) had been tested for antibodies against Toxoplasma gondii during pregnancy. Women attending primary antenatal care at a specialist in obstetrics and gynaecology were most likely to be tested. Among women with no signs of prior toxoplasmosis, 35% (110/317) were tested only once. DISCUSSION: The majority of pregnant women are being tested for toxoplasmosis even though such testing has no documented preventive effect and has not been recommended. PMID- 14738654 TI - The quality of routine examinations of the newborn performed by midwives and SHOs; an evaluation using video recordings. AB - OBJECTIVE: To evaluate the quality of the routine examination of the newborn as carried out by senior house officers (SHOs) and midwives. DESIGN: Randomised controlled trial set in a district general hospital in Southeast England. Eligible babies were randomised to a midwife (11) or an SHO (8) who were then video recorded undertaking the routine newborn examination. MAIN OUTCOME MEASURES: Quality of 62 observed technical and communication components of the newborn examination, as agreed between independent consultant paediatrician and senior midwife raters. RESULTS: Major differences were found in the rated quality of examinations between midwives and SHOs for the examinations of the heart and lungs, for the overall quality of the examination, and in areas of communication skills. Quality of examination of the hips was assessed as poor for both professional groups. Where there were significant differences between examiners, the quality of the midwives' examinations was higher. Inter-rater agreement between the consultant paediatricians and senior midwives ranged from excellent to poor for different items of the examination with a mean kappa value of 0.42 across all items indicating moderate agreement. CONCLUSIONS: The quality of midwife examinations exceeded that of SHOs. All midwives who examine receive formalized training in the examination of the newborn; SHOs may benefit from similar specific training. Training for both professional groups may need to be reviewed, particularly in relation to agreement on 'gold standards' for the screening of developmental dysplasia of the hip. PMID- 14738655 TI - Neonatal birth parameters of positive newborns at PKU screening as predictors of false-positive and positive results at recall-testing. AB - OBJECTIVES: To investigate the predictive role that neonatal birth parameters of positive newborns at phenylketonuria (PKU) screening have on false-positive and positive results. We reviewed 195 newborns (115 males and 80 females) that had false-positive results between 1998 and 2001. A total of 4386 randomly selected neonates (2191 males and 2195 females) who tested normal at the first investigation in the same period, were used as negative-controls. A total of 38 PKU neonates (17 males and 21 females) diagnosed between 1990 and 2001 were used as positive-controls. METHODS: Phenylalanine concentration was measured with a fluorometric multitask plate counter Wallac 1420 VICTOR F (Perkin Elmer, Finland) and the fluorescent ninhydrine method (EG&G Wallac neonatal phenylalanine kit) using a recall cut-off level >120 micromol/l (2 mg/dl) of phenylalanine on dried blood spots. A multivariate logistic regression analysis was performed to evaluate the predictive role that body parameters (sex, gestational age, parity, weight, length and head circumference) of positive newborns at PKU screening had on false-positive and positive results at recall PKU tests. RESULTS: The risk of false-positive results is higher (~48%) in females than in males. Moreover, for each 100g of body weight reduction, the risk of false positive is around 4.2% higher. The risk of confirmation increased by 39% per week of gestational age. CONCLUSIONS: In conclusion, our results suggest that preterm or low-birth-weight neonates recalled at the first investigation are more likely to be due to false positives, whereas the risk of confirmation is higher in at-term neonates. By implication, the phenylalanine cut-off value for premature or low-body-weight infants could be higher. PMID- 14738656 TI - The burden of prevention: downstream consequences of Pap smear testing in the elderly. AB - CONTEXT: Although cervical cancer is an unusual cause of death among women 65 and older, most elderly women in the US report continuing to undergo periodic Pap smear screening. OBJECTIVE: To describe the incidence of Pap smears and downstream testing among elderly women. SETTING: Claims-based analysis of female Medicare enrollees age 65 and older. METHODS: Using three years of Medicare Part B 5% Files (1995-1997), we differentiated between women undergoing screening Pap smears and those undergoing Pap smears for surveillance of previous abnormalities or Pap smear follow-up. We determined the proportion of elderly women undergoing Pap smear testing and rates of downstream testing and procedures after an initial Pap smear. RESULTS: Four million female Medicare beneficiaries over 65 years underwent Pap smear testing between 1995 and 1997, representing 25% of the eligible population. After adjusting for underbilling for Pap smears under Medicare, 43% of women over 65 are estimated to have undergone Pap smear testing during the 3-year period. The large majority (90%) of Pap smears were for screening, while 10% were done for surveillance or follow-up. For every 1000 women with a screening Pap smear, 39 had at least one downstream intervention within eight months of the initial Pap smear, including seven women who underwent colposcopy and two women who had other surgical procedures. Rates of downstream interventions were considerably higher for women undergoing Pap smear follow-up (302 per 1000 with at least one downstream intervention), and surveillance of previous abnormalities (209 per 1000 with a downstream intervention). CONCLUSION: Cervical cancer screening is widespread among elderly American women, and follow up testing is not uncommon, particularly among the ten percent of women who appear to be in a cycle of repeated testing. This substantial volume of testing occurs despite the rarity of cervical cancer deaths and unknown benefits of screening in this age group. PMID- 14738657 TI - Risk of invasive cervical cancer after three consecutive negative Pap smears. AB - OBJECTIVES: To determine the factors that influence risk of cervical cancer after three consecutive negative Pap smears. METHODS: A cohort study was conducted using data from the British Columbia Cervical Cancer Screening Program and British Columbia Cancer Registry. Analysis was based on a one percent sample of women aged 20-69 years with Pap smears enriched with all invasive cervical cancer cases diagnosed between 1994-99. Screening intervals, after three negative screens, were created with the following variables: age at beginning of interval, interval length, previous cytologic abnormality and previous cervical procedure. The risk of cervical cancer by histologic type was calculated using survival analysis methods. RESULTS: The sample consisted of 10,509 women, who contributed 28,309 intervals, and 371 cervical cancer cases. The incidence rate of invasive squamous cervical cancer increased with time since last screen up to six years. Women with a history of dysplasia remained at elevated risk for squamous cancer, hazard ratio=2.6 (95% confidence interval [CI]=1.9, 3.4) but age or previous procedure were not related to risk. No relationship between time since last screen and non-squamous cancer risk was found although history of a previous procedure was significant. The marginal effectiveness of Pap smears declined with increasing frequency of use. CONCLUSIONS: This study confirmed the preventive effect of Pap smear screening and its dependency on frequency of use. Women with a history of dysplasia, prior to three consecutive negatives, were at increased risk of developing invasive squamous cervical cancer compared with women with no such history. PMID- 14738658 TI - An assessment of smear taker training in the Northern & Yorkshire cervical screening programme. AB - OBJECTIVE: To assess the impact of a regional smear taker training scheme within the Northern and Yorkshire Cervical Screening Programme. DESIGN: An analysis of cervical screening KC activity returns to the Department of Health before and after the introduction of a structured training programme for smear takers. SETTING: Northern and Yorkshire Region of England. SUBJECTS: Women participating in cervical screening. RESULTS: The introduction of a region-wide smear taker training programme has resulted in fewer inadequate cervical smears, fewer opportunistic smears and fewer women aged below 20 having cervical smears. The training programme is cost effective. CONCLUSIONS: It is recommended that all cervical screening programmes consider the introduction of smear taker training. It is a cost-effective approach to running the cervical screening programme. PMID- 14738659 TI - Screening for liver cancer: results of a randomised controlled trial in Qidong, China. AB - OBJECTIVES: To investigate the effectiveness of screening for liver cancer in reducing mortality from the disease in a high-risk population in China. SETTING: A randomised controlled trial was carried out among men aged 30-69 who were chronic carriers of hepatitis-B virus (HBsAg positive) during the period 1989 1995 in Qidong county, Jiangsu Province, China. METHODS: 5581 HBsAg carriers were identified by population screening and randomly assigned to a screening group (group A, 3712 men), and controls (group B, 1869 men). Screening was planned to be six monthly alpha-fetoprotein (AFP) assays, with follow-up of subjects having an abnormal (>/=20 micrograms/l) test. All subjects were followed up for liver cancer and/or death until 31 December 1995. RESULTS: The overall sensitivity and specificity of the programme was 55.3% and 86.5%, respectively; in subjects who complied with all scheduled screening tests, the values were 80.0% and 80.9%. Three hundred and seventy-four primary liver cancer (PLC) cases were diagnosed. The percentage of cases in stage I was significantly higher in group A (29.6%) than in group B (6.0%). The one-, three-, and five-year relative survival rates were 23.7%, 7.0%, and 4.0% in group A, and 9.7%, 4.0%, and 4.1% in group B respectively, with no difference in five-year survival between the groups. The mortality rate in the screened group (1138 per 100,000 person-years) was not significantly different from that in the controls (1114 per 100,000). A Poisson regression model showed that the probability of death (rate ratio) in the screening group was 0.83 (95% CI 0.68-1.03) relative to the control group. CONCLUSIONS: Screening with AFP resulted in earlier diagnosis of liver cancer, but the gain in lead time did not result in any overall reduction in mortality, because therapy for the patients found by screening was ineffective. Further studies using improved methods of screening, diagnosis and treatment are indicated. PMID- 14738660 TI - British Chief Medical Officer recommendations for genital chlamydial infection screening may not be applicable outside the UK. PMID- 14738663 TI - Fatal pulmonary embolism after bariatric operations for morbid obesity: a 24-year retrospective analysis. AB - BACKGROUND: Pulmonary embolism (PE) is a leading cause of death following gastric bypass operations for morbid obesity. Although its incidence appears to be stable, the number of bariatric operations performed annually is increasing considerably; hence, the isolated fatal PE is no longer a rare occurrence. The records of patients undergoing bariatric surgical operations since 1979 were reviewed to determine specific factors that increased the risk of developing a fatal PE. Both recommended and optional indications for prophylactic inferior vena cava (IVC) filter placement in patients considered at high risk were also examined. MATERIALS AND METHODS: Between September, 1979 and March, 2003, 5,554 operations were performed for clinically severe obesity. These operations included jejuno-ileal bypass, horizontal gastroplasty, Roux-en-Y gastric bypass with a 30-cc pouch, modified biliopancreatic diversion, the Sapala-Wood Micropouch gastric bypass (Micropouch(SM)), Lap-Band, and revisions. 12 fatal pulmonary emboli (0.21 %) were identified. All but 1 embolus occurred within 30 days following surgery. RESULTS: In 11 of 12 patients, at least 1 co-morbidity known to increase the risk of postoperative venous thromboembolism (VTE) was identified. 4 co-morbidites were common to 4 patients (33%): venous stasis disease (VSD), BMI >/= 60, truncal obesity, and obesity hypoventilation syndrome/sleep apnea syndrome (OHS/SAS). 6 of 12 patients (50%) had a BMI >/= 60. Another 6 had chronic leg swelling with stasis dermatitis. 2 patients experienced a previous PE, and 1 patient reported a history of deep vein thrombosis (DVT). CONCLUSION: 4 patients (33%) demonstrated a combination of risk factors (VSD, BMI >/= 60, truncal obesity, OHS/SAS) recognized as significant for the development of postoperative VTE. In such patients, prophylactic IVC filter placement is highly recommended. Filter placement for other factors, such as age, body build, hypercoagulable state, etc., should be considered on an individual basis. PMID- 14738664 TI - Bariatric surgery in adolescence. is this the best age to operate? AB - BACKGROUND: Bariatric surgery in morbidly obese adolescents is controversial. Many argue that morbidly obese individuals should be of adult age before undergoing bariatric operations, despite the progressive and debilitating course of this increasingly common disease. MATERIALS AND METHODS: 19 consecutive adolescent patients, aged 13-17, underwent vertical banded gastroplasty-Roux-en-Y gastric bypass between May 1990 and August 2001. Average BMI was 49 kg/m(2), range 38-67. All had one or more co-morbidities. Follow-up was obtained up to 10 years. RESULTS: Postoperative BMI at the maximum time of follow-up, mean 5.5 years (range 1-10 years), was 28 (range 23 to 45). Only one patient did not lose enough weight and was considered a failure. There were two revisions and no mortality or morbidity. All co-morbidities disappeared. Family and patients were pleased with the surgery. CONCLUSIONS: Early surgical intervention should be offered to a greater number of adolescents to minimize the emotional and physical consequences of morbid obesity. PMID- 14738665 TI - Depression score predicts weight loss following Roux-en-Y gastric bypass. AB - BACKGROUND: The prevalence of obesity is increasing in the United States. Bariatric surgery is the only intervention that can reliably induce and maintain significant weight loss in obese patients. The association between pre-surgical severity of depression and success at weight loss following Roux-en-Y gastric bypass (RYGBP) has not yet been fully elucidated. METHODS: 145 charts of patients who underwent RYGBP for morbid obesity were reviewed. 47 patients who filled out the Beck Depression Inventory (BDI) before surgery and completed 1 year of follow up were studied. The relationship between pre-surgical severity of depression and success at weight loss was examined through multivariate regression analysis using percent excess weight loss (%EWL) as a dependent variable and BDI score as one of the predictors. RESULTS: Weight loss at 1 year was significantly related to the BDI score before surgery (P =0.014). BDI score was also found to be a significant predictor of the amount of weight lost (kg) 1 year after surgery (P =0.027). Age (P =0.03) and initial body mass index (BMI) (P =0.011) were the only other variables with significant independent relations to %EWL. CONCLUSIONS: Our data show a positive correlation between pre-surgical severity of depression as measured by BDI score and the 1-year success at weight loss after RYGBP as measured by %EWL. More depressed individuals tend to lose greater amounts of weight compared with less depressed individuals. Future prospective studies should examine possible mechanisms and effects of depression and other psychiatric disturbances on long-term weight loss after RYGBP. PMID- 14738666 TI - Clinical experience using the computerized digital stapling system in open gastric bypass surgery for morbid obesity. AB - BACKGROUND: Roux-en-Y gastric bypass is currently the preferred method of surgical treatment for morbid obesity. Complications associated with this technique include anastomotic leaks and anastomotic strictures. Our technique employs a computerized digital stapling system (SurgASSIST). This stapling system has been utilized in affecting both the end-to-end gastrojejunostomy and end-to side enteroenterostomy anastomoses. METHODS: SurgASSIST is a computerized stapling system consisting of a power console and a 2.13-meter flexshaft. Using a remote control, various stapling cartridges can be opened and closed, and the tip of the flexshaft can be steered in four directions prior to firing. Because the firing mechanism and force are generated away from the surgical field, torque applied to the anastomotic site is minimized. The system is designed to approximate tissue to the correct staple height, based on tissue thickness, which precludes excessive compression of the tissues. RESULTS: We have employed the SurgASSIST computerized digital stapling system in a series of 403 cases with 558 applications. We have experienced no anastomotic leaks. There were 7 strictures (1.7%) requiring balloon dilatation. There were no strictures requiring surgical intervention. CONCLUSION: Our extensive clinical experience using the SurgASSIST computerized digital stapling system has shown it to be a safe and effective method for performing stapled anastomoses. PMID- 14738667 TI - Management of acute bleeding after laparoscopic Roux-en-Y gastric bypass. AB - BACKGROUND: The authors reviewed the incidence of hemorrhage after laparoscopic Roux-en-Y gastric bypass (LRYGBP). The purpose of this study was to determine the incidence of this complication and to evaluate various treatment options. MATERIAL AND METHODS: The records for 450 consecutive patients who had undergone LRYGBP over a 30-month period, were retrospectively reviewed. In all patients, the abdominal cavity had been drained with 2 19-Fr closed suction drains. The charts of patients who had developed an intraluminal or an intraabdominal bleed were chosen for further review. RESULTS: 20 patients (4.4%) developed an acute postoperative hemorrhage. The bleeding was intraluminal in 12 cases (60%), manifested by a drop in hematocrit, tachycardia and melena. The other 8 patients (40%) developed intra-abdominal hemorrhage, confirmed by large bloody output from the drains. 3 patients (15%) with intraluminal bleeding were unstable and required a reoperation. All others were successfully treated with observation, and 15 patients (75%) required blood transfusions. CONCLUSIONS: The diagnosis and treatment of acute intraluminal bleeding after LRYGBP represents a surgical dilemma, mainly due to the inaccessibility of the bypassed stomach and the jejuno jejunostomy, as well as the risks associated with early postoperative endoscopy. The presence of large intra-abdominal drains allows for bleeding site localization (intraluminal vs intraabdominal) and for more accurate monitoring of the bleeding rate. Most cases respond to conservative therapy. Failure of conservative management of intraluminal bleeding, however, is more problematic and may require operative intervention. A treatment algorithm is proposed. PMID- 14738668 TI - DaVinci robotic-assisted laparoscopic bariatric surgery: is it justified in a routine setting? AB - BACKGROUND: Laparoscopic silicone adjustable gastric banding (SAGB) has gained popularity for the surgical treatment of morbid obesity. The implantable gastric stimulator (IGS) system represents a novel surgical alternative. We aimed to assess the feasibility of robotic-assisted laparoscopic bariatric operations and to critically elucidate the technical and financial advantages and patient outcome. METHODS: Robotic-assisted laparoscopic bariatric procedures were performed on 10 consecutive patients using the daVinci robot system (4 SAGB, 4 IGS, 2 SAGB revisions). 10 conventional laparoscopic-operated patients (4 SAGB, 4 IGS, 2 SAGB revisions) during the learning curve served as controls. Equipment, operative technique and procedural time were evaluated. A cost analysis was calculated. RESULTS: The personnel equipment, numbers of trocars and operation technique were comparable in both groups. The mean operative time was 137 min (range 110-175) and 97 min (60-140) in robotic-assisted and conventional laparoscopy, respectively (P =0.04). Establishment of the pneumoperitoneum and placement of trocars and robotic arms took a mean of 30 min (15-45) in the robotic-assisted group, compared with 5 min in the control group (P <0.001). In 1 patient, intraoperative gastric injury was suspected and led to band removal in the robotic-assisted group. There was no postoperative complication. Average procedural costs were significantly higher in the robotic-assisted group. CONCLUSION: Primary and revisional robotic-assisted bariatric surgery is technically simple, with the benefit of precise instrument handling. However, it is still expensive, the set-up of the system is time-consuming, and a limited variety of instruments are available presently. PMID- 14738669 TI - Prospective randomized comparison of linear staplers during laparoscopic Roux-en Y gastric bypass. AB - BACKGROUND: The development of laparoscopic linear staplers has enabled minimally invasive approaches to bariatric surgery, but there have been no comparison studies of the two current 6-row devices. We report our experience with a prospective randomized comparison of 6-row linear staplers during laparoscopic Roux-en-Y gastric bypass (LRYGBP). METHODS: From January to March 2003, 100 patients were randomly assigned to undergo LRYGBP with either an Endo-GIA Universal 6-row stapler (USSC) or the ETS-Flex 6-row stapler (Ethicon). Mean preoperative BMI was 49+/-8 for 50 Endo-GIA patients, and 49+/-7 for 50 ETS-Flex patients. Parameters measured included quantity of cartridges, handles, hemoclips, estimated blood loss, misfires, OR time, postoperative leaks and bleeds, and cost. RESULTS: Mean follow-up was 135 days (range 90- 180). The ETS Flex group experienced significantly more misfires (28% vs 2%, P <.001), hemoclips applied (30+/-9 vs 21+/-7, P <.001), estimated blood loss (132+/-56 vs 100+/-32 ml, P <.001) and OR time (66+/-19 vs 58+/-13 mins, P <.02) compared with the Endo-GIA group respectively. There was one postoperative leak associated with the ETS-Flex group and two postoperative bleeds with the Endo-GIA group, which were not a significant differences. The Endo-GIA group averaged $319 more per case for staple cost. CONCLUSION: While the ETS-Flex stapler was less expensive, it was associated with more technical failures requiring surgeon intervention to reduce potential patient morbidity, compared with the Endo-GIA. PMID- 14738671 TI - Early experience with two-stage laparoscopic Roux-en-Y gastric bypass as an alternative in the super-super obese patient. AB - BACKGROUND: Surgical management of the supersuper obese patient (BMI >60 kg/m2) has been a challenging problem associated with higher morbidity, mortality, and long-term weight loss failure. Current limited experience exists with a two-stage biliopancreatic diversion and duodenal switch in the supersuper obese patient, and we now present our early experience with a two-stage gastric bypass for these patients. METHODS: We completed a retrospective bariatric database and chart review of super-super obese patients who underwent laparoscopic sleeve gastrectomy as a first-stage procedure followed by laparoscopic Roux-en-Y gastric bypass as a second-stage for more definitive treatment of obesity. RESULTS: During a two-year period, 7 patients with BMI 58-71 kg/m2 underwent a two-stage laparoscopic Roux-en-Y gastric bypass by two surgeons at the Mount Sinai Medical Center. 3 patients were female, 4 patients were male, and the average age was 43. Prior to the sleeve gastrectomy, the mean weight was 181 kg with a BMI of 63. Average time between procedures was 11 months. Prior to the second-stage procedure, the mean weight was 145 kg with a BMI of 50 and average excess weight loss of 37 kg (33% EWL). Six patients have had follow-up after the second-stage procedure with an average of 2.5 months. At follow-up the mean weight was 126 kg with a BMI of 44 and average excess weight loss of 51 kg (46% EWL). The mean operative times for the two procedures were 124 and 158 minutes respectively. The average length of stay for all procedures was 2.7 days. 4 patients had 5 complications, which included splenic injury, proximal anastomotic stricture, left arm nerve praxia, trocar site hernia, and urinary tract infection. There were no mortalities in the series. CONCLUSIONS: Laparoscopic sleeve gastrectomy with second-stage Roux-en-Y gastric bypass are feasible and effective procedures based on short-term results. This two-stage approach is a reasonable alternative for surgical treatment of the high-risk supersuper obese patient. PMID- 14738672 TI - The use of simvastatin for the prevention of gallstones in the lithogenic prairie dog model. AB - BACKGROUND: Surgery for morbid obesity is rapidly increasing. Patients undergoing bariatric surgery are prone to gallstone development during the rapid weight loss. These patients are often given medications such as ursodeoxycholic acid to prevent gallstone formation; however, these medications are often poorly tolerated by patients, who subsequently discontinue them. We performed a study in a lithogenic animal model to assess the effectiveness of a potential alternate medication for gallstone prevention. METHODS: 20 male prairie dogs were randomly separated into 2 groups and fed a lithogenic diet for 28 days. The study group animals were given 2.5 mg of the HMG-CoA reductase inhibitor simvastatin. Total cholesterol and triglycerides were measured and an open cholecystectomy was performed on each animal at the conclusion of the study period. The gallbladder was visually inspected for gallstones and microscopic biliary cholesterol crystal formation. RESULTS: There was a decrease of 36% in the total cholesterol of the study animals compared to controls. The animals treated with simvastatin showed gallstone formation in 5/10 (50%) of animals, compared with 6/10 (60%) of control animals. The study animals demonstrated microscopic cholesterol crystal formation in 80%, identical to the number found in the control animals. CONCLUSION: Despite a reduction in cholesterol, simvastatin prevented neither gallstone formation nor biliary cholesterol crystals in this animal model. Given the rapid increase in the number of bariatric surgical procedures coupled with the poor tolerance of ursodeoxycholic acid, viable alternatives should continue to be sought for these patients. PMID- 14738673 TI - Effects of weight loss on QT interval in morbidly obese patients. AB - BACKGROUND: Obesity causes structural changes to the heart that may influence its function. Furthermore, morbid obesity is associated with an acquired prolongation of the QTc interval that may lead to potentially hazardous arrhythmias. The present study investigated the effect of body weight loss following vertical banded gastroplasty (VBG) on the QTc interval. METHODS: 17 morbidly obese patients, scheduled for elective VBG, were studied before the operation and 8-10 months postoperatively, when each patient had achieved a weight loss of >/= 25% of the preoperative body weight. RESULTS: 15 patients achieved significant body weight loss of >/= 25% within the first 8-10 postoperative months (P <0.001). This weight loss, corresponding to an excess weight loss of 48.7% and a mean body mass index (BMI) reduction from 49.7 kg/m2 to 36.6 kg/m2, was followed by significant shortening of the QTc interval from 428 msec to 393 msec (P <0.001). CONCLUSIONS: The significant postoperative weight loss following VBG was accompanied by shortening of the QTc interval. This effect is expected to reduce the incidence of fatal conditions associated with the long QT syndrome, such as malignant ventricular arrhythmias and sudden death, and therefore improve morbidity and mortality. PMID- 14738674 TI - Plasma leptin levels after vertical banded gastroplasty for morbid obesity: effects of an acidified meal. AB - BACKGROUND: Leptin is considered one of the anorectic messengers to the central nervous system in lean subjects. Although it is secreted by the gastric mucosa, there are contradictory evidences of its involvement in mediating the acute satiety effect of the meal in obese patients. The effects of restrictive operations on meal-stimulated leptin secretion are unknown. METHODS: The effects of a standard acidified (pH 3) meal on leptin release were investigated in obese patients, before and after vertical banded gastroplasty (VBG). 8 morbidly obese patients (BMI 49.1+/-6.5) had serum leptin determination after an overnight fast. Samples were taken basally and every 30 minutes after the meal for 3 hours. The test was repeated after 20% BMI reduction. 5 lean volunteers (BMI 22.5+/-1.7) served as the control group. RESULTS: In obese patients, basal serum leptin fell from 62+/-20.4 to 23.8+/-15.7 ng/ml after the operation (P <0.01) but still with significant differences vs the control group (5.6+/-3 ng/ml). The meal was associated with a significant decrease of serum leptin (ANOVA test, P <0.01), and significant differences between obese patients after surgery and lean subjects were found. CONCLUSION: Serum leptin was reduced by the meal in obese patients and VBG did not attain satiety through serum leptin changes. PMID- 14738675 TI - Effect of significant intermediate-term weight loss on serum leptin levels and body composition in severely obese subjects. AB - BACKGROUND: Leptin, produced by adipose tissue, signals body fat content to the hypothalamus. Serum leptin levels (SLL), elevated in obese humans, decrease with weight loss. This study investigated the reduction of SLL and fat mass following restrictive bariatric surgery. METHODS: Obese subjects (body mass index [BMI] >35 kg/m2, n=154) undergoing gastric banding (weight-reduced subjects) were investigated for SLL and body composition before surgery and for 2 years after. Overweight subjects matched for fat mass and gender (fat mass-matched overweight controls, n=194) and subjects who had never been obese (normal weight controls, n=158) were studied for comparison. RESULTS: SLL were highest in weight-reduced subjects and decreased with weight loss (P <0.001), remaining elevated compared with normal weight controls (P <0.001) but lower than fat mass-matched overweight controls (women: P <0.04). At 2 years, SLL normalized for fat mass (allowing comparison between various levels of adiposity) were lower in weight-reduced subjects compared with fat mass-matched overweight controls (women: P =0.003), yet were similar for weight-reduced subjects at 2 years compared with normal weight controls despite 14 kg greater fat mass. Relative lean mass of extremities in weight-reduced subjects increased with weight loss (P <0.001). CONCLUSION: SLL decreased after considerable weight loss more than could be accounted for by fat mass or BMI reduction alone. This disproportionate decrease in SLL might point to a mechanism that evolved as adaptation to starvation during times of famine. Thus, post-obese subjects may be at risk of weight-regain due to disproportionately low SLL and increased appetite via the leptin-melanocortin pathway. PMID- 14738676 TI - Plasma ghrelin concentratin in the short-term following biliopancreatic diversion. AB - BACKGROUND: Ghrelin is a hormone that increases food intake in rodents and in humans. After gastric bypass surgery, a marked decrease in circulating ghrelin levels has been observed, and it was suggested that this may contribute to the weight-reducing effect of gastric bypass. In this study, the changes in circulating ghrelin levels following biliopancreatic diversion (BPD) were investigated. MATERIALS AND METHODS: Serum ghrelin concentration was measured in obese patients prior to and 5 days and 2 months following BPD. RESULTS: At the short-term following BPD, marked reduction of serum ghrelin was observed, while thereafter the values returned to initial levels. CONCLUSION: Unlike following reducing diet or gastric bypass, following BPD only an initial reduction of serum ghrelin concentration was observed, while at 2 months following the operation, when food intake had nearly completely resumed, the values returned to the preoperative levels. This is consistent with the hypothesis that ghrelin production from the stomach is greatly influenced by the direct contact of ingested food with the gastric cells. PMID- 14738677 TI - Calcium and vitamin D depletion and elevated parathyroid hormone following biliopancreatic diversion. AB - BACKGROUND: Biliopancreatic diversion (BPD) is associated with a 70% excess weight loss (EWL) at 10 years, but there are concerns regarding long-term nutritional sequelae. Metabolic bone disease has been documented following Roux en-Y gastric bypass. METHODS: Patients who underwent a BPD from 1998 to 2001 were studied. A questionnaire was designed to review BPD patients and collect information on weight loss, frequency of gastrointestinal disturbances and compliance with multivitamin recommendations. The review included a blood test for vitamin D, parathyroid hormone (PTH), alkaline phosphatase (ALP) and calcium. RESULTS: Of the 82 patients who underwent BPD during this period, the median %EWL at 36 months was 73.0%. 75.6% suffered diarrhea. At median follow-up of 32 months (18-50), 25.9% of patients were hypocalcemic, 50% had low vitamin D, 23.8% had elevated ALP, and 63.1% had elevated PTH, despite 82.9% taking multivitamins. CONCLUSION: BPD results in significant weight loss. However, 1 in 4 patients are hypocalcemic, and 1 in 2 have a low vitamin D, despite multivitamin supplementation. BPD patients require routine calcium and vitamin D supplementation for life. Long-term sequelae from these abnormal serum levels are not known. PMID- 14738678 TI - Achalasia in the context of morbid obesity: a rare but important association. AB - BACKGROUND: The simultaneous occurrence of achalasia and morbid obesity is rare. Nevertheless, the surgical therapy of morbid obesity may be harmful, if undiagnosed achalasia were left untreated. We report the clinical presentation and response to treatment of achalasia in the context of morbid obesity. METHODS: From 1998 to 2002, 638 patients underwent surgery for morbid obesity. Preoperative upper gastrointestinal radiography was performed in all patients. Three patients had manometric confirmation of achalasia. The characteristic symptoms were recurrent episodes of regurgitation, chronic cough and aspiration. No patient reported dysphagia or recent weight loss. RESULTS: All patients had a duodenal switch procedure and in two a concurrent Heller myotomy was added. The other patient required a Heller myotomy after a duodenal switch had been performed, because the motility study was initially misinterpreted. All patients reported gradual resolution of presenting symptoms after myotomy. CONCLUSIONS: A careful symptomatic history focusing on aspiration, regurgitation and cough may identify the unusual combination of achalasia and morbid obesity. Treatment of morbid obesity alone may lead to progression of pulmonary symptoms. PMID- 14738679 TI - Radiology for laparoscopic adjustable gastric banding: a simplified follow-up examination method. AB - BACKGROUND: The aim of this study was to identify factors which allow a reliable, standardized and simplified approach for the radiologic follow-up of obese patients who have undergone surgery for laparoscopic adjustable gastric banding (LAGB) placement. METHODS: A study was made of 370 consecutive single-contrast upper gastrointestinal series in 159 consecutive patients (122 women, 31 men; mean age 40.6 years; mean body weight 135.8 kg) who had undergone LAGB for morbid obesity and were symptomatic or had inadequate weight loss. The control group consisted of 38 subjects who had undergone LAGB, had satisfactory weight loss and were asymptomatic at follow-up. RESULTS: Each gastric portion above the band was satisfactorily evaluated by measuring its vertical diameter. The optimal projection for measurement of stoma size was predicted before administration of the contrast agent and was achieved when the band was visualized in profile. The clinical pictures were not always related to stoma size. Surgical complications were gastric herniation (8 patients, twice in 1 patient, 5.7%), malpositioned band (1 patient, 1.6%), port twisting (13 patients, 8.2%), and disconnection or leakage of the device (8 patients, 5.0%). CONCLUSION: Essential criteria for the radiologic evaluation of LAGB are: position of port and tubing; stoma size; and volume of each upper gastric portion. PMID- 14738680 TI - Optimization of access-port placement for the lap-band system. AB - BACKGROUND: The technical aspects of access-port (AP) placement are not generally described in Lap-Band series. METHODS: From November 2000 to April 2002, we performed Lap-Band procedures laparoscopically on 180 patients. A retrospective review was conducted of 3 consecutive AP-placement techniques in nonselected and demographically identical groups. In Group A (n=48, Nov. 2000 to July 2001), the AP was placed at the left subcostal margin. In Group B (n=23, Aug. 2001 to Sept. 2001), the AP was tunneled over the subcostal fascia towards the subxiphoid area. In Group C (n=109, Oct. 2001 to Apr. 2002), the AP tubing was tunneled over the subcostal fascia and connected to the AP, which was inserted through a 3-cm subxiphoid incision. RESULTS: AP-related problems occurred within the first few months following surgery. In Group A, 24 of the APs (50%) were tilted, and 14 (29%) were completely flipped over. 11 APs (23%) were found to be broken. 19 patients (40%) underwent an additional AP-related procedure. In group B, 12 APs (52%) were tilted and 1 patient required surgery to turn the AP. In Group C, 8 APs (7%) were turned slightly. 1 AP was found to be broken and required surgery to replace it. In this group, all APs were accessible for adjustment in the office. CONCLUSIONS: Tunneling the AP along the left subcostal area is an important technique to protect the AP system from breakage, by changing AP-tube position from vertical to horizontal in relation to abdominal wall movement. This technique also keeps the AP-tube connection over the fascia and protects it from "wear and tear" forces. The addition of fixation at the subxiphoid location helps maintain a straight orientation of the AP for easier adjustments. PMID- 14738681 TI - Band leakage after laparoscopic adjustable gastric banding. AB - BACKGROUND: Laparoscopic adjustable gastric banding is effective in inducing weight loss, as well as being minimally invasive, totally reversible, and adjustable to the patient's needs. Nevertheless, leakage of the adjustable balloon is a known complication. The aim of this study was to assess the incidence and reasons for balloon leakage of the Swedish adjustable gastric band (SAGB). PATIENTS AND METHODS: Between January 1996 and December 2002, 566 patients (475 women, 91 men) underwent a laparoscopic SAGB implantation. Two groups of patients were analyzed: patients with early postoperative leakage (Group E) and patients with late postoperative leakage (Group L). All data (age, gender, pre- and postoperative weight, time of weight gain, band filling status) were prospectively collected in a computerized data bank. For the detection of gastric band leakage, radiography and the technetium-99m colloid scintigraphy was used. RESULTS: 25 band leakages were observed in 22 patients (4.4%). All these patients had a silent presentation of band leakage, with weight regain and an ability to eat without major restriction. Band leakages in group E were detected during the band filling period after a median follow-up of 8 months and after 30.3 months (P <0.0001) in group L. In group E, all 13 leakages possibly resulted from inappropriate handling of the device during surgery. In 2 cases in group L, a tear of the balloon had occurred where it is fixed to the band. The other 10 bands showed breaks at the edges of the inner side of the balloon. All leakages could be detected by (99m)Tc colloid scintigraphy, whereas only 58% of the leakages could be detected by radiography. CONCLUSION: Band leakage is a rare complication and should be considered if a patient starts to regain weight. Operative failure as well as material defects may account for this complication. The balloon leakage can effectively be detected by (99m)Tc colloid-scintigraphy. PMID- 14738682 TI - Initial experience with Swedish adjustable gastric band at Al-noor hospital. AB - BACKGROUND: The aim of this 4-year study is to give an initial account of the weight loss and complications after applying the Swedish adjustable gastric band (SAGB) at Al-Noor Specialist Hospital-Makkah. MATERIALS AND METHODS: 97 morbidly obese patients underwent SAGB. RESULTS: 94 patients benefited by losing weight. 1 patient had a minor complication (wound infection) and 1 did not return for follow-up. 2 early patients died of pulmonary embolism. The 64 females (68%) and 30 males had mean BMI 50.8 kg/m2 before SAGB and 35.0 at 2 years after SAGB. CONCLUSION: Gastric banding generally was a valuable procedure for morbidly obese patients. It was easy to perform, with low operative risk and few longterm complications. PMID- 14738683 TI - Clinical significance of central obesity in laparoscopic bariatric surgery. AB - BACKGROUND: Laparoscopic surgery had increased the interest and growth of bariatric surgery. Whether central obesity has any adverse effect in laparoscopic bariatric surgery is not clear. METHODS: 612 morbidly obese patients received laparoscopic bariatric surgery,in a prospectively controlled clinical trial of the outcome of the bariatric surgery. For comparison, subjects were dichotomized into either a central obesity group or peripheral obesity group, based on waist/hip ratio (WHR). Various biochemistry and blood count variables, and perioperative and postoperative results were measured. RESULTS: There were more female (458) than male patients (154). Male patients had higher BMI, and female patients were younger. 56 of 154 male patients (36.4%) belonged to the central obesity (WHR >1.0), and 321 of 458 female patients (70.1%) had central obesity (WHR >0.85). Central obesity was associated with age but not with BMI in males. In females, central obesity was associated with increased BMI. Central obesity predicted increased hyperglycemia and triglyceride levels in both male and female patients. Male patients with central obesity had higher WBC counts than the other patients. Although central obesity was associated with more intra-operative blood loss and prolonged recovery in female patients, there was no increase in complication rate or difference in postoperative weight loss. CONCLUSION: Central obesity is associated with a higher degree of hyperglycemia, hyperlipidemia and leukocytosis in morbidly obese patients who undergo bariatric surgery. Although there is increased technical difficulty in patients with severe central obesity, laparoscopic bariatric surgery is safe and effective in producing weight loss. PMID- 14738684 TI - Bariatric surgery and bariatric psychology: general overview and the Dutch approach. AB - BACKGROUND: Obesity is a chronic, multifactorially caused disease with serious somatic and psychosocial comorbidity as well as economical consequences. In the Netherlands, between 1993 and 1997, the prevalence of morbid obesity was 0.2% for men and 0.6% for women. Although bariatric surgery generally is an effective intervention, it does not lead to equal results in every patient. The long-term efficacy is predominantly determined by compliance to adequate dietary rules in which psychosocial factors can play a major role. METHODS: Questionnaires were sent to the surgery departments of all hospitals in the Netherlands. Subsequently, a second questionnaire was sent to clinical psychology departments of hospitals which perform bariatric surgery. RESULTS: In 28 Dutch hospitals (19%), bariatric surgery is being performed, mostly using restrictive procedures. Almost all hospitals have a multidisciplinary selection-process, and all surgeons and psychologists use multiple selection-criteria. Regarding these criteria, there is more consensus between surgeons than between psychologists. In most hospitals, patients are psychologically assessed prior to surgery. However, postoperative assessment is relatively rare, as is preoperative and postoperative psychological treatment. CONCLUSION: In the Netherlands, bariatric surgery is still relatively uncommon and mostly limited to restrictive procedures. Irrespective of BMI and eating behavior, the majority of patients will be offered a restrictive procedure. The involvement by the psychological and/or psychiatric discipline is not optimal yet; especially, postoperative assessment and pre- and postoperative treatment are not frequently performed, in spite of the fact that these programs can enhance the success rate of bariatric surgery. PMID- 14738685 TI - Use of anti-emetics after intragastric balloon placement: experience with three different drug treatments. AB - BACKGROUND: Tropisetron treatment was compared with alizapride treatment. The secondary aim was to assess whether droperidol supplement would still improve the therapeutic outcome of tropisetron. MATERIALS AND METHODS: A series of 51 obese patients was treated with an intragastric balloon to obtain weight reduction. Patients were divided at random into 3 groups. Each group received a different antiemetic and spasmolytic regimen to control postoperative nausea and vomiting for 24 hours. Statistical analysis of both parameters showed that all 3 populations are comparable and the studied incidence of vomiting was only influenced by the choice of the antiemetics used. A specially developed form was completed during the recovery period every 6 hours until 24 hours postoperatively and recorded all episodes of vomiting. The incidence of vomiting was then calculated as number of episodes/24 hours RESULTS: The incidence of vomiting was significantly lower in the tropisetron group compared to the alizapride group. There was no significant difference between the tropisetron group and the tropisetron plus droperidol group. CONCLUSION: To decrease the incidence of vomiting in patients undergoing intragastric balloon placement, tropisetron proved to be the most effective antiemetic. A supplement of droperidol gave no better results but impaired postoperative mood and wellbeing. Alizapride was least effective. PMID- 14738686 TI - Ruptured appendicitis after laparoscopic Roux-enY gastric bypass: pitfalls in diagnosing a surgical Abdomen in the morbidly obese. AB - A recent gastric bypass can mask the symptoms of an acute abdomen. Physical examination is generally unreliable and subtle clinical symptoms or signs should alert clinicians to a significant postoperative problem. In morbidly obese patients, the presence of overt peritoneal findings is usually ominous, leading to sepsis, organ failure and death. We report a case of ruptured appendicitis following a laparoscopic Roux-en-Y gastric bypass. The patient developed tachycardia, fever, and leukocytosis in the absence of abdominal pain or positive upper GI contrast studies. Eventually, a CT scan revealed a large pelvic abscess and inflammation. A subsequent exploratory laparotomy confirmed a perforated appendicitis with pelvic peritonitis. Her recovery was rapid and uneventful. This case highlights the pitfalls in promptly diagnosing an unrelated acute surgical abdomen postoperatively in the morbidly obese patient. The need for extreme vigilance and a low threshold for aggressive intervention in the period after bariatric surgery is emphasized. PMID- 14738687 TI - Postoperative rhabdomyolysis with bariatric surgery. AB - Rhabdomyolysis has been reported in all postoperative patients including those in prone, supine, lithotomy and lateral decubitus positions. Only a few reports suggest that bariatric surgical patients are at risk for rhabdomyolysis. We describe a male (BMI 69 kg/m2) who underwent an uneventful open Roux-en-Y gastric bypass for weight reduction lasting 5 hours. Postoperatively the patient suffered oliguria. Evaluation included subjective pain in both hips, a normal temperature and physical examination, creatinine increase to 3.5 mg/dl, CPK levels as high as 41,000 IU/L, and urinalysis showing a large amount of occult blood with 5-7 RBCs/HPF. Intravenous hydration with 0.9% normal saline, bicarbonate, and mannitol demonstrated initial success, but the patient eventually developed renal failure, respiratory distress, and tachycardia leading to cardiac arrest. Prior to his death, intraoperative evaluation demonstrated intact anastomoses. Obese patients undergoing bariatric surgery should be considered at risk for rhabdomyolysis, especially in view of prolonged surgeries, difficult physical examination, low volume status, and larger or immobile patients. PMID- 14738688 TI - Massive pericardial effusion following a laparoscopic adjustable gastric banding. AB - Laparoscopic adjustable gastric banding (LAGB) is currently the most common bariatric surgical procedure. The most frequent complications of LAGB surgery are pouch dilatation, gastric prolapse, band erosion, stoma obstruction and access port problems. We report a rare life-threatening case of massive pericardial effusion as a complication of an infected Lap-Band. The management of this condition included subxiphoid pericardial window, removal of the band and later conversion to gastric bypass. PMID- 14738689 TI - Gastric bezoar complicating laparoscopic adjustable gastric banding, and review of literature. AB - Gastric bezoars may be formed in the normal stomach as a result of foreign body consecrations of various objects with inability to pass through the pylorus. Classically, most bezoars occur as a complication of gastric surgery which creates a low acid environment, decreased peristalsis, and abnormal pyloric function. Bariatric surgery has been associated with a low incidence of bezoar formation. However, to date there has been no documentation of bezoars occurring after laparoscopic adjustable gastric banding, which is one of the surgical options available for the treatment of morbid obesity. We report a case of a gastric bezoar that occurred 8 months after gastric banding. PMID- 14738690 TI - Green urine after intragastric balloon placement for the treatment of morbid obesity. AB - BACKGROUND: The intragastric balloon is filled with saline and methylene blue dye, to detect balloon deflation early and prevent bowel obstruction, by monitoring the patient's urine for changes in color. METHODS: An intragastric balloon filled with 590 ml of saline plus 10 ml of methylene blue was endoscopically placed under sedation in a 22-year-old man with morbid obesity (BMI 42 kg/m2). 3 days later, the patient's urine changed to dark green, and, suspecting a leaking balloon, endoscopy was repeated under sedation. RESULTS: No signs of balloon deflation were seen, and the urine returned to normal color. The next day, the urine turned green again. 7 days later, the urine discoloration finally disappeared. CONCLUSION: Propofol, a sedative commonly used by anesthesiologists during endoscopic procedures, is known to have several side effects, and urine discoloration is one of them, albeit rare. This benign side effect must be known to obesity surgeons to avoid pointless medical expenditure, unnecessary balloon removal and distress for patients and clinicians. PMID- 14738691 TI - Measuring outcomes following bariatric surgery: weight loss parameters, improvement in co-morbid conditions, change in quality of life and patient satisfaction. AB - Restrictive and particularly malabsorptive bariatric operations achieve significant sustained weight loss. Results from different operations have been difficult to compare. The aims of this review are: 1) to indicate the limitations of outcomes reported as weight-related parameters; 2) to document some of the patient characteristics that impact weight loss; 3) to assess the literature documenting improvement in obesity-related medical conditions; and 4) to review studies that quantitate changes in health-related quality of life (QoL). Weight related parameters such as body mass index and % excess weight inconsistently correlate with body fat. Direct determination of body fat with bioelectric impedance may offer more reliable outcome parameters. Patient characteristics such as gender, age, weight, body mass index, ethnicity, race and socioeconomic status affect weight loss following bariatric operations. Improvements in co morbid conditions are poorly documented in many studies. Standardized instruments that assess health-related QoL have shown differing values. SF-36 has given inconsistent results following bariatric operations. Both BAROS and IWQoL-Lite have demonstrated significant improvements after surgery. Bariatric surgeons have rarely used patient satisfaction as an outcome parameter. This review suggests that bariatric operations should be judged by change in fat mass or fat mass index, improvement in obesity-related medical conditions, change in health related QoL as judged by standardized instruments, and level of patient satisfaction. In addition, surgeons should characterize their study population and report outcomes for sub-populations. PMID- 14738692 TI - The BAROS and the Moorehead-Ardelt quality of life questionnaire. PMID- 14738693 TI - Limb measurements in duodenal switch. PMID- 14738696 TI - Intermittent explosive disorder and impulsive aggression: the time for serious study is now. PMID- 14738697 TI - Amphetamine salt compound treatment for adults with attention deficit hyperactivity disorder. PMID- 14738698 TI - Late-life depression in the primary care setting. PMID- 14738699 TI - Evidence-based medicine and geriatric psychiatry. AB - The vast amount of clinical information available today and the difficulties that clinicians have in applying this information have given rise to interest in evidence-based medicine. An additional impetus for the development of the evidence-based medicine movement has been the increasing recognition that much of medical practice is not evidence-based. This article presents a brief overview of the principles of evidence-based medicine and its historical development, discusses some common criticisms of evidence-based practice, and illustrates its application to address a clinical problem. Suggestions are also provided for clinicians interested in developing evidence-based medicine skills. PMID- 14738700 TI - Assessing and predicting functional impairment in Alzheimer's disease: the emerging role of frontal system dysfunction. AB - Alzheimer's disease (AD) is associated with neuropsychologic and neuropsychiatric dysfunction and is a leading cause of disability among the elderly. Impairments in activities of daily living (ADL) contribute significantly to the disability reported among patients with AD and diminish quality of life for patients and their families. ADL assessment represents an important component of the diagnosis, tracking, and management of AD. Further, an understanding of the determinants of ADL dysfunction is critical for the early identification of individuals at risk for functional disability and for improved patient care. This manuscript reviews methods for assessing ADL in patients with AD and summarizes the available literature on the neuropsychologic and neuropsychiatric correlates of functional impairment in AD. The emerging role of frontal system dysfunction as an important determinant of ADL impairment is discussed, and recommendations for clinical practice and future research are provided. PMID- 14738701 TI - The significance of subsyndromal depression in geriatrics. AB - Clinically significant non-major depression has been underinvestigated, despite its high prevalence and public health impact. Although there is increasing recognition of the importance of subsyndromal forms of depression, their nosologic boundaries and neurobiologic mechanisms remain largely unknown. This review discusses the literature pertaining to the current concepts, phenomenology, neurobiology, and treatment approaches for geriatric non-major clinically significant depression. The authors examine the similarities and differences between various subtypes of depressive disorders, and compare non major clinically significant depression in the elderly with non-geriatric adult populations. The authors draw conclusions from the published literature and present clinical criteria for the diagnosis of clinically significant non-major depression in the elderly. PMID- 14738703 TI - Divalproex for cluster B personality disorders. PMID- 14738704 TI - Borderline personality disorder therapy with omega-3 fatty acids. PMID- 14738702 TI - Current status of psychotherapy for mental disorders in the elderly. AB - Research on psychotherapy for older adults with psychiatric disorders has demonstrated its effectiveness, although the majority of research has been conducted on major depression. Recent advances in extending this research to additional diagnostic categories and treatment settings are reviewed. Psychotherapy appears promising in the treatment of minor depression, dysthymia, anxiety disorders, depression with comorbid personality disorders, depression with comorbid cognitive impairment, and as an adjunctive treatment in psychotic disorders. Psychotherapy also has been successfully transported to the primary care setting, and shows potential in long-term care and in-home settings. Many of these studies are preliminary, however; additional research is needed with larger, more diverse samples across a variety of psychiatric diagnoses and treatment settings. PMID- 14738706 TI - Depression and personality disorder. AB - Research into the relationship between depression and personality disorder is compromised by a number of methodologic factors, including differing concepts of personality disorder, the validity of a personality disorder diagnosis, the effect of mood on diagnosis, and the overlap between some personality disorder symptoms and mood symptoms. Personality pathology is common in patients with personality disorder. Reasons for this include a "scar" effect of chronic low mood on attitudes and behaviors, as well as possible risk factors via certain personality traits. The negative effect of comorbid personality disorder on treatment outcome in depressed patients may be less than previously believed. Possible reasons include treatment bias in non-controlled trials and the increasing use of selective serotonin reuptake inhibitors rather than tricyclic antidepressants. Many personality traits and disorders may be part of the psychopathology of depression and share a common origin. PMID- 14738705 TI - New developments in the neurobiology of borderline personality disorder. AB - Within the past several years, research on the clinical phenomena and neurobiology of borderline personality disorder has increased substantially. Borderline personality disorder is currently best thought of in terms of dimensions rather than as a categorical disorder. This article reviews the most recent findings on two of the core dimensions--affective dysregulation and impulsivity. Most of the neuropsychologic, physiologic, endocrinologic, and neuroimaging data support the theory that a dual brain pathology, affecting prefrontal and limbic circuits, may underlie this hyperarousal-dyscontrol syndrome. PMID- 14738707 TI - Compulsive disorders. AB - Compulsive disorders include a diverse group of conditions characterized by excessive thoughts or preoccupations combined with poorly controlled behaviors. They include trichotillomania, kleptomania, pathologic gambling, compulsive buying disorder, compulsive sexual behavior, and compulsive computer use. Some investigators have suggested that these conditions constitute a spectrum of disorders linked to obsessive-compulsive disorder. Others have questioned the validity of this conceptualization, and have debated the relationship between these disorders. Nevertheless, much has been learned about compulsive disorders, and there have been some successes with psychotherapeutic and psychopharmacologic treatments. Recent therapy-based interventions have moved from psychodynamic treatments toward cognitive-behavioral modalities. Serotonin reuptake inhibitors remain the best-studied pharmacologic treatment, but researchers have also explored other antidepressants, opioid agonists, mood stabilizers, and atypical antipsychotics. PMID- 14738708 TI - Update on pharmacotherapy of borderline personality disorder. AB - Pharmacotherapy is a very common form of treatment for borderline personality disorder or its concomitant disorders. This paper reviews all the open-label and placebo-controlled trials of second generation medications studied in samples of well-defined borderline patients. Most of the medications studied in double blind, placebo-controlled trials were efficacious. Most of these medications were also useful in treating symptoms of affective dysregulation and impulsive aggression, which have been suggested to be the core dimensions of psychopathology of underlying borderline personality disorder. Taken together, the results of these studies suggest that the choice of medication can be guided as much by tolerability and safety as by symptom presentation. It also suggests that the common practice of polypharmacy, which has no empiric support, may be unnecessary for most patients with borderline personality disorder. PMID- 14738709 TI - Gender differences in personality traits and disorders. AB - Personality disorders differ in prevalence by gender. The most striking findings concern antisocial personality disorder, which is more common in men, and borderline personality disorder, which is more common in women. These differences are not artifacts, but reflect gender differences in the personality traits that underlie Axis II diagnoses. PMID- 14738710 TI - Chemotherapy induced gastrointestinal toxicity in rats: involvement of mitochondrial DNA, gastrointestinal permeability and cyclooxygenase -2. AB - PURPOSE: The gastrointestinal damage induced by xenobiotics is occurring more frequently and with greater toxicological significance than previously thought. Although there are some preliminary clinical studies and reports, there does not appear to be an extensive examination of gastrointestinal toxicity of various chemotherapeutic agents in the rat. This study was undertaken to examine the suitability of a rat model to detect the gastrointestinal damage after administration of various anti-neoplastic agents including etoposide, teniposide, melphalan, 5-fluorouracil, methotrexate and cisplatin. METHODS: Acute toxic doses of indomethacin and chemotherapeutic agents were administered to rats. The urinary excretion of orally administered sucrose and 51(Cr)-EDTA were measured as markers of gastroduodenal and intestinal permeability, respectively. Cyclooxygenase-2 messenger RNA and mitochondrial DNA damage were measured as toxicological endpoints. RESULTS: Each anti-neoplastic agent examined induced appreciable and significant dose-dependent increase in gastrointestinal permeability that correlated with gross toxicological and pathological changes to the gastrointestinal tract including ulceration and bleeding. COX-2 mRNA was upregulated > 2 fold in intestinal mucosa with enteropathy and dose-dependent mitochondrial oxidative damage was apparent in gastric and intestinal mucosa. After administration of each drug, the rats presented with histological evidence of drug-induced gastroenteropathy, ulceration and increased cecal hemoglobin. CONCLUSIONS: The rat appears to be a suitable model to study gastrointestinal toxicity of chemotherapeutic agents and non-steroidal anti-inflammatory drugs. Damage to mitochondrial DNA occurs in both the gastric and intestinal epithelium after the administration of these agents and may be an important factor in the pathogenesis and resolution of gastrointestinal toxicity. PMID- 14738711 TI - Permeation prediction of M100240 using the parallel artificial membrane permeability assay. AB - PURPOSE: Kansy et al first introduced the Parallel artificial membrane permeation assay (PAMPA) in 1998. In this system, the permeability through a membrane formed by a mixture of lecithin and an inert organic solvent on a filter support is assessed. PAMPA shows definite trends in the ability of molecules to permeate membranes by transcellular passive diffusion. Its simplicity, low cost, high throughput, and wide pH range make it very attractive in modern drug discovery. Based on this concept, Whohnsland et al, Sugano et al and Zhu et al modified the assay and used it to screen compound permeability. We used PAMPA for the permeation prediction of M100240, which was unable to be determined by cell-based assays due to compound instability. METHODS: In this study, 92 commercially available agents provided the structural diversity used to generate a mathematical prediction model for human fraction absorbed, M100240--an acetate thioester of MDL 100,173. Permeation of M100240 and MDL 100,173 was evaluated using the parallel artificial membrane permeability assay (PAMPA). The donor and recipient solutions consisted of 0.5N HCl (pH 1.5) or phosphate-buffered saline (pH 5.5 or 7.4) with 2% dimethyl sulfoxide. The donor solution also contained 200 mM M100240 or MDL 100,173. RESULTS: M100240 had a medium permeation at pH 5.5 (2.99%), corresponding to a high predicted Fa in humans (92%). Permeation of MDL 100,173 was low at this pH (0.72%), corresponding to a medium-to-low predicted Fa (46). At pH 7.4, the permeation of M100240 was low (approximately 1%) and no permeation was apparent for MDL 100,173. CONCLUSIONS: We predicted M100240 is likely to be well absorbed via passive diffusion across the human gastrointestinal tract following oral administration. PMID- 14738712 TI - Preparation and in vitro transfection efficiency of chitosan microspheres containing plasmid DNA:poly(L-lysine) complexes. AB - PURPOSE: Studies on DNA complexes with cationic polymers are prompted by the search for nonviral DNA carriers for gene therapy. Among them, poly(L-Lysine) (PLL) has been extensively studied. On the other hand, these systems deliver DNA as a bolus without long-term release. The aims of this study were to encapsulate plasmid DNA:poly(L-lysine) (pDNA:PLL) complexes into chitosan microspheres as an alternative to the PLL based gene delivery and investigate its in vitro release and transfection characteristics as well as plasmid DNA integrity and stability against serum and DNase I challenge. METHODS: pUC18 plasmid DNA that encoded beta galactosidase was used as a model. The microspheres were prepared by complex coacervation method and the release and in vitro transfection properties were investigated. pDNA:PLL complexes were prepared at two different mass ratios. In vitro release studies were performed at 37 +/- 0.5 degrees C and drug release was monitored both spectrophotometrically and fluorometrically. Structural integrity of the pDNA:PLL complexes were determined by Southern blotting analysis. Protective effect of encapsulation of pDNA:PLL complexes against DNase I and serum treatment were also studied. In vitro transfection studies were performed by using 3T3 cell line. RESULTS: According to our in vitro release data, the mass ratio of pDNA:PLL significantly affected the release of pDNA:PLL complexes from chitosan microspheres, and the structure of the plasmid DNA did not change during the experiments. pDNA:PLL-loaded chitosan microspheres indicated high stability against fetal bovine serum and DNase I treatment for a week. In vitro transfection data showed that pDNA:PLL-loaded chitosan microspheres could be effectively transfected 3T3 cells in vitro. CONCLUSION: As a conclusion, pDNA:PLL complexes could be encapsulated into chitosan microspheres with maintaining their structural and functional integrity and this system may be a good alternative for polycation based gene carriers. PMID- 14738713 TI - Protection of pancreatic beta-cell by the potential antioxidant bis-o hydroxycinnamoyl methane, analogue of natural curcuminoid in experimental diabetes. AB - PURPOSE: To evaluate the antioxidant defense by bis-o-hydroxycinnamoylmethane, analogue of the naturally occurring curcuminoid bis-demethoxycurcumin in streptozotocin induced diabetes in male Wistar rats and its possible protection of pancreatic beta-cell against gradual loss under diabetic condition. METHODS: Male wistar rats were divided into five groups. Group1 served as control rats. Group2 was control rats treated intragastrically with bis-o-hydroxycinnamoyl methane at a dose of 15 mg/kg body weight for 45 days. Group3, 4 and 5 rats were injected with 40 mg /kg body weight of streptozotocin to induce diabetes. Group4 rats were treated with the drug similar to group2 and group5 rats treated with the reference drug glibenclamide intragastrically for a similar period. After 45 days, the levels of plasma glucose, glycated hemoglobin, enzymic antioxidants (SOD, CAT) and non-enzymic antioxidants Vit C, Vit E was determined. Histopathological sections of the pancreas were examined. RESULTS: The levels of plasma glucose and glycated hemoglobin which were elevated in group3 diabetic rats were reduced after treatment with the drug. The antioxidant levels showed an increase in the case of treated diabetic rats as compared to group3 diabetic rats. The islets were shrunken in group3 diabetic rats in comparison to normal rats. In the treated diabetic rats there was expansion of islets. CONCLUSIONS: The experimental drug bis-o-hydroxycinnamoylmethane enhances the antioxidant defense against reactive oxygen species produced under hyperglycemic conditions and thus protects the pancreatic b -cell against loss and exhibits antidiabetic property. PMID- 14738714 TI - Influence of storage conditions and type of plasticizers on ethylcellulose and acrylate films formed from aqueous dispersions. AB - PURPOSE: To investigate the influence of storage conditions and types of plasticizers on the properties and stability of ethylcellulose and polymethacrylate films and to elucidate the mechanism for the changes observed. METHODS: Films were prepared from Surelease, Aquacoat and Eudragit L 30D dispersions by the casting method. The effects of different plasticizers on the morphology, transparency, mechanical property and water vapour permeability of the prepared films were studied. The film samples were exposed to storage conditions of 30 degrees C and 50 or 75 %RH. Samples were removed at pre determined time intervals for mechanical testing and analysis of plasticizer content in the films. RESULTS: It was found that films prepared from aqueous ethylcellulose dispersions were relatively weaker and more brittle than acrylate films. Acrylate films did not show any significant change in mechanical property when stored at high humidity. However, the properties of ethylcellulose films stored at high humidity varied depending on the type of plasticizers present. CONCLUSIONS: The changes in mechanical property of ethylcellulose films on storage were mainly attributed to the loss of plasticizers during storage, causing further coalescence of ethylcellulose films and to a smaller extent, reduction in moisture content of the film. PMID- 14738715 TI - Application of a new high performance liquid chromatography method to the pharmacokinetics of dibudipine in rats. AB - PURPOSE: To develop a HPLC method for assay of dibudipine in biological fluids and to study its pharmacokinetics in the rat. METHODS: HPLC: 2 microl (20 microg/ml) mebudipine as internal standard, 0.2 ml NaOH 1 M and 2 ml ethyl acetate were added to 0.2 ml of rat plasma. The mixture was shaken for 10 min, centrifuged, and the supernatant was dried under nitrogen. The dissolved residue was injected to a C18 analytical column. Mobile phase flowed at 1 ml/min with a composition of methanol--water-acetonitrile (70-25-5). The eluent was monitored at 238 nm. Pharmacokinetic study: plasma samples were collected periodically after intravenous (0.5 mg/kg) or oral (10 mg/kg) administration of dibudipine to rats (n = 4/group). In addition, separate groups of animals were administered 0.5 mg/kg doses of the drug for serial collection of brain, heart, kidney and liver (n = 4/time). The concentration of the drug in tissue or plasma was assayed using the above HPLC method. RESULTS: Calibration curves were linear over a concentration of 10-1000 ng/ml and CV was less than 10%. Dibudipine showed a bi exponential decline after IV injection in the rats with a t1/2 beta of 2.5 +/- 0.5(mean +/- SE) hr. Oral bioavailability was low. Distribution of dibudipine to the examined tissues was rapid, and with the exception of the brain, the concentrations of the drug in all tissues were higher than the plasma levels CONCLUSIONS: The HPLC method was simple and convenient. Moreover, it could be applied to investigations of the pharmacokinetics of dibudipine in the rat. PMID- 14738716 TI - A comparison of gastrointestinal permeability induced by diclofenac-phospholipid complex with diclofenac acid and its sodium salt. AB - PURPOSE: Gastrointestinal (GI) side effects of the nonsteroidal anti-inflammatory drug (NSAID) diclofenac may be reduced if it is administered as a complex with phospholipid. The upper and lower GI permeability induced by a diclofenac dipalmitoyl phosphatidyl choline (DPPC) complex were compared with those of diclofenac acid and its sodium salt in rats. METHODS: Pharmacokinetic studies were carried out to assess bioavailability of diclofenac preparations. Adult male Sprague-Dawley rats were dosed orally (equivalent to 15 mg/kg diclofenac sodium) as the acid or its sodium salt as well as diclofenac-DPPC complex. Upper and lower GI permeability, as surrogate markers of toxicity were determined using sucrose and 51Cr-EDTA, respectively. RESULTS: At 1 h post-dose only diclofenac sodium induced a significant increased upper GI permeability. Three h post-dose all formulations significantly increased upper GI permeability although the diclofenac acid had the least effect. In the lower GI tract, the induced increase in permeability was significant at 1 and 3 h post-dose for all formulations with no significant differences between them. CONCLUSION: The induced upper and lower GI toxicity of diclofenac was formulation and time dependent. The lack of effect of diclofenac acid was due to the decreased availability of the drug. In the upper GI tract, up to 1 h post-dose, the diclofenac-DPPC complex demonstrated reduced upper gastroduodenal permeability as measured by sucrose. However, the protective effect of DPPC did not last and was not extended to the lower GI tract due to the systemic effect, contribution from the enterohepatic recirculation and/or dissociation of the complex. In assessing diclofenac GI toxicity, the effect of the different formulations on the entire GI tract at various times after drug administration must be considered. PMID- 14738717 TI - The antinociceptive activities of 1-(4-aryl-2-thiazolyl)-3,5-disubstituted-2 pyrazolines in mouse writhing test. AB - PURPOSE: Sympathetic nervous system stimulation, which releases noradrenalin, influences the nociceptive activity that develops after tissue injury. The alpha2 adrenergic agonist, clonidine, produces analgesia through a central mechanism but also inhibits noradrenalin release at terminal nerve fiber endings. METHODS: In this study the effects of some 1-(4-aryl-2-thiazolyl)-3,5-disubstituted-2 pyrazolines (compounds 1a-1e) as analogues of clonidine with some modifications were studied by writhing test, a visceral pain model in mice. RESULTS: Compounds 1a and 1c induced significant reduction in writhing response when compared to control. Regarding the dose-response relationship of compounds 1a and 1c, it is evident that the activity of these compounds is in direct proportion to their dose and all compounds show activities comparable to clonidine. In addition compounds 1a and 1c, having methoxy and nitro groups respectively at para position of 4-aryl showed better antinociception than compounds having similar groups in meta position namely 1b and 1e. This might be due to better interaction of these compounds with the alpha2-receptors. CONCLUSIONS: The antinociceptive properties of these newly synthesized compounds is comparable to clonidine. Further studies are needed to explore the differences in the efficacy and safety of synthesized compounds. PMID- 14738718 TI - Allergy to corticosteroids: update and review of epidemiology, clinical characteristics, and structural cross-reactivity. PMID- 14738719 TI - Factors influencing the induction phase of skin sensitization. PMID- 14738720 TI - Contact dermatitis education in dermatology residency programs: can (will) the American Contact Dermatitis Society be a force for improvement? AB - BACKGROUND: Contact dermatitis accounts for a considerable portion of outpatient clinic visits to dermatologists. The state of education in contact dermatitis at the level of dermatology residency training in the United States has not been examined. OBJECTIVE: To assess the state of education in contact dermatitis in dermatology residency programs in the United States. METHOD: Cross-sectional survey of directors and chief residents of 105 dermatology training programs accredited by the American College of Graduate Medical Education. RESULTS: Seventy-seven percent of directors and 74% of chief residents responded to the survey. In general, both sets of respondents gave concordant responses although responses from directors were more positive. With respect to didactic education, the vast majority of programs (> 73%) held lecture conferences on contact dermatitis. Less than one-third included contact dermatitis-focused journals in journal club conferences. A bare majority of programs (57% of directors, 53% of chief residents) identified a faculty expert in contact dermatitis, with almost all experts conducting patch-test clinics and providing lectures on contact dermatitis. Seventy-five percent of experts were members of the American Contact Dermatitis Society (ACDS). Although residents in most programs (> 78%) performed patch tests to diagnose contact dermatitis, there were 14 programs in which none of their graduates performed such tests. Moreover, only 27% of programs had rotations dedicated to contact dermatitis and/or patch testing. Finally, directors and chief residents predicted that most graduates will incorporate the TRUE Test and not the more extensive or customized patch tests in their practices. CONCLUSIONS: Several opportunities for improving contact dermatitis education in residency programs were identified, including recruitment or development of more faculty experts in contact dermatitis, creation of rotations dedicated to contact dermatitis, and greater inclusion of contact dermatitis focused journals in journal club conferences. As the principal interest group for contact dermatitis in the United States, the ACDS is the logical organization to spearhead improvement of contact dermatitis education in residency programs. PMID- 14738721 TI - Contact dermatitis in adolescents. AB - BACKGROUND: Adolescents between the ages of 10 and 19 are exposed to a series of substances capable of causing contact dermatitis. OBJECTIVES: (1) To study the frequency and characteristics of allergic contact dermatitis in adolescents, (2) to characterize the group being studied, and (3) to verify the main sensitizing substances among this age group. METHODS: From 1996 to 2001, 1,027 patients with a suspicion of contact dermatitis were analyzed, and patients between 10 and 19 years of age were selected. These patients were submitted to contact tests. RESULTS: Among the 102 adolescents, 93 (91%) were female and 9 (9%) were male. The face was the area most affected by dermatosis. The contact tests were positive in 64 patients (56%), whereas in 45 (44%) they were negative. The main location of the contact dermatitis was the face (36%). The substances with higher frequencies of sensitization were nickel sulfate in 33 (31%) patients and tosylamide-formaldehyde resin in 13 (12%) patients. CONCLUSION: Contact dermatitis in adolescents was more frequent in white girls and on the face. The substances with greater frequency of sensitization were nickel sulfate and tosylamide-formaldehyde resin. These two substances are related to adolescent habits and behavior. PMID- 14738722 TI - Disperse dyes in fabrics of patients patch-test-positive to disperse dyes. AB - BACKGROUND: Little evidence exists that dyes to which individuals are patch test positive are in those garments that they suspect cause their dermatitis, which makes diagnosis and management of colored-textile allergic contact dermatitis difficult. OBJECTIVE: We determined whether disperse dyes to which a patient suspected of having a colored garment-dye contact allergy were in the garment that the patient suspected to be the cause of his or her skin lesions. METHODS: Each patient was patch-tested with 12 disperse dyes in a commercial patch test series. Disperse dyes in the submitted garment(s) were identified. The dyes to which each patient was patch test positive were compared with the dyes identified in that patient's submitted fabric(s). When a dye appeared in both lists, a correlation was found. RESULTS: Twenty-two of 32 garments received contained disperse dyes. In all, 35 different disperse dyes were identified. Twelve dyes elicited a reaction in at least one patient. Only nine patients were patch test positive to at least one dye identified in the fabric submitted. Dyes appearing in both lists were Blue 106 (8 times), Blue 124 (2 times), Yellow 3 (once), and Red 1 (once). CONCLUSION: Dyes to which a patient was patch test positive were infrequently identified in the fabric suspected to be the cause of the skin lesions, which means that the next step will be to patch-test with fabric swatches and extracts of dye from the submitted garments. PMID- 14738723 TI - Agave contact dermatitis. PMID- 14738724 TI - Reimbursement for patch testing at academic centers: the Achilles' heel of academic contact dermatitis specialists. PMID- 14738725 TI - Occupational allergic contact dermatitis in an obstetrics and gynecology resident. PMID- 14738726 TI - Painful dermatitis of the fingertip. PMID- 14738727 TI - Lidocaine hydrochloride. PMID- 14738728 TI - Uncoupling of brain activity from movement defines arousal States in Drosophila. AB - BACKGROUND: An animal's state of arousal is fundamental to all of its behavior. Arousal is generally ascertained by measures of movement complemented by brain activity recordings, which can provide signatures independently of movement activity. Here we examine the relationships among movement, arousal state, and local field potential (LFP) activity in the Drosophila brain. RESULTS: We have measured the correlation between local field potentials (LFPs) in the brain and overt movements of the fruit fly during different states of arousal, such as spontaneous daytime waking movement, visual arousal, spontaneous night-time movement, and stimulus-induced movement. We found that the correlation strength between brain LFP activity and movement was dependent on behavioral state and, to some extent, on LFP frequency range. Brain activity and movement were uncoupled during the presentation of visual stimuli and also in the course of overnight experiments in the dark. Epochs of low correlation or uncoupling were predictive of increased arousal thresholds even in moving flies and thus define a distinct state of arousal intermediate between sleep and waking in the fruit fly. CONCLUSIONS: These experiments indicate that the relationship between brain LFPs and movement in the fruit fly is dynamic and that the degree of coupling between these two measures of activity defines distinct states of arousal. PMID- 14738729 TI - Protein kinase D-mediated anterograde membrane trafficking is required for fibroblast motility. AB - BACKGROUND: Locomoting cells exhibit a constant retrograde flow of plasma membrane (PM) proteins from the leading edge lamellipodium backward, which when coupled to substrate adhesion, may drive forward cell movement. However, the intracellular source of these PM components and whether their continuous retrograde flow is required for cell motility is unknown. RESULTS: To test the hypothesis that the anterograde secretion pathway supplies PM components for retrograde flow that are required for lamellipodial activity and cell motility, we specifically inhibited transport of cargo from the trans-Golgi network (TGN) to the PM in Swiss 3T3 fibroblasts and monitored cell motility using time-lapse microscopy. TGN-to-PM trafficking was inhibited with a dominant-negative, kinase dead (kd) mutant of protein kinase D1 (PKD) that specifically blocks budding of secretory vesicles from the TGN and does not affect other transport pathways. Inhibition of PKD on the TGN inhibited directed cell motility and retrograde flow of surface markers and filamentous actin, while inhibition of PKD elsewhere in the cell neither blocked anterograde membrane transport nor cell motile functions. Exogenous activation of Rac1 in PKD-kd-expressing cells restored lamellipodial dynamics independent of membrane traffic. However, lamellipodial activity was delocalized from a single leading edge, and directed cell motility was not fully recovered. CONCLUSIONS: These results indicate that PKD-mediated anterograde membrane traffic from the TGN to the PM is required for fibroblast locomotion and localized Rac1-dependent leading edge activity. We suggest that polarized secretion transmits cargo that directs localized signaling for persistent leading edge activity necessary for directional migration. PMID- 14738730 TI - A spectraplakin is enriched on the fusome and organizes microtubules during oocyte specification in Drosophila. AB - BACKGROUND: During Drosophila oogenesis a membranous organelle called the fusome has a key function in the establishment of oocyte fate and polarity, ultimately leading to the establishment of the major body axes of the animal. The fusome is necessary for the microtubule-driven restriction of markers of oocyte fate to the oocyte, but the mechanism by which the fusome organizes the microtubules is not known. RESULTS: We have identified the spectraplakin Short stop (Shot) as a new component of the fusome. Spectraplakins are giant cytoskeletal linker proteins, with multiple isoforms produced from each gene. Shot is the sole spectraplakin in Drosophila. The phenotype caused by the absence of Shot is not similar to that of other components of the fusome but instead is similar to the absence of the downstream components that interact with microtubules: the dynein/dynactin complex-associated proteins Egalitarian and BicaudalD. Shot is required for the association of microtubules with the fusome and the subsequent specification of the oocyte in 16-cell cysts. Shot is also required for the concentration of centrosomes into the oocyte, a process thought to be independent of microtubules because it still occurs in the presence of microtubule depolymerizing drugs. This suggests that Shot may protect some microtubules from depolymerization and that these microtubules are sufficient for this process. CONCLUSIONS: Shot provides the missing link between the fusome and microtubules within meiotic cysts, which is essential for the establishment of the oocyte. Shot associates with the fusome and is required for microtubule organization. We suggest that it does this directly, via its microtubule binding GAS2 domain. PMID- 14738731 TI - Genes required for systemic RNA interference in Caenorhabditis elegans. AB - RNA interference (RNAi) in the nematode worm, Caenorhabditis elegans, occurs systemically. Double-stranded RNA (dsRNA) provided in the diet can be absorbed from the gut lumen and distributed throughout the body, triggering RNAi in tissues that are not exposed to the initial dsRNA trigger. This is in marked contrast to other animals, in which RNAi does not spread from targeted tissues to neighboring cells. Here, we report the characterization of mutants defective in the systemic aspect of RNAi, but not in the core RNAi process itself. Analysis of these mutants suggests that dsRNA uptake is a specific process involving several unique proteins. PMID- 14738732 TI - The human premotor cortex is 'mirror' only for biological actions. AB - Previous work has shown that both human adults and children attend to grasping actions performed by another person but not necessarily to those made by a mechanical device. According to recent neurophysiological data, the monkey premotor cortex contains "mirror" neurons that discharge both when the monkey performs specific manual grasping actions and when it observes another individual performing the same or similar actions. However, when a human model uses tools to perform grasping actions, the mirror neurons are not activated. A similar "mirror" system has been described in humans, but whether or not it is also tuned specifically to biological actions has never been tested. Here we show that when subjects observed manual grasping actions performed by a human model a significant neural response was elicited in the left premotor cortex. This activation was not evident for the observation of grasping actions performed by a robot model commanded by an experimenter. This result indicates for the first time that in humans the mirror system is biologically tuned. This system appears to be the neural substrate for biological preference during action coding. PMID- 14738733 TI - Early vision impairs tactile perception in the blind. AB - Researchers have known for more than a century that crossing the hands can impair both tactile perception and the execution of appropriate finger movements. Sighted people find it more difficult to judge the temporal order when two tactile stimuli, one applied to either hand, are presented and their hands are crossed over the midline as compared to when they adopt a more typical uncrossed hands posture. It has been argued that because of the dominant role of vision in motor planning and execution, tactile stimuli are remapped into externally defined coordinates (predominantly determined by visual inputs) that takes longer to achieve when external and body-centered codes (determined primarily by somatosensory/proprioceptive inputs) are in conflict and that involves both multisensory parietal and visual cortex. Here, we show that the performance of late, but not of congenitally, blind people was impaired by crossing the hands. Moreover, we provide the first empirical evidence for superior temporal order judgments (TOJs) for tactile stimuli in the congenitally blind. These findings suggest a critical role of childhood vision in modulating the perception of touch that may arise from the emergence of specific crossmodal links during development. PMID- 14738734 TI - Condensin regulates rDNA silencing by modulating nucleolar Sir2p. AB - The tandem array of ribosomal DNA (rDNA) in Saccharomyces cerevisiae is subjected to transcriptional silencing of RNA polymerase II-transcribed genes. This form of silencing depends on SIR2 and has been tightly linked to the suppression of rDNA recombination and the control of cellular lifespan. Paradoxically, rDNA silencing takes place in the context of an extremely high rate of RNA polymerase I transcription. Because rDNA silencing requires different factors than HMR and telomere silencing, the chromatin structure and the mechanisms of silencing must be fundamentally different. Here we show that yeast condensin organizes the specialized topology of rDNA chromatin. We then demonstrate that this function is necessary for maintaining the correct balance of telomeric and nucleolar Sir2p. Condensin mutants relocalize telomeric Sir2p to rDNA and show histone hyperacetylation at telomeres. Our data reveal the implication of yeast condensin in the arrangement of rDNA repeats into a heterochromatic-like structure that is important for the correct delineation of silencing domains in the nucleus. PMID- 14738735 TI - The vertebrate Ndc80 complex contains Spc24 and Spc25 homologs, which are required to establish and maintain kinetochore-microtubule attachment. AB - How kinetochores bind to microtubules and move on the mitotic spindle remain unanswered questions. Multiple systems have implicated the Ndc80/Hec1 (Ndc80) kinetochore complex in kinetochore-microtubule interaction and spindle checkpoint activity. In budding yeast, Ndc80 copurifies with three additional interacting proteins: Nuf2, Spc24, and Spc25. Although functional vertebrate homologs of Ndc80 and Nuf2 exist, extensive sequence similarity searches have not uncovered homologs of Spc24 and Spc25. We have purified the xNdc80 complex to homogeneity from Xenopus egg extracts and identified two novel interacting proteins. Although the sequences have greatly diverged, we have concluded that these are the functional homologs of the yeast Spc24 and Spc25 proteins based on limited sequence similarity, common coiled-coil domains, kinetochore localization, similar phenotypes, and copurification with xNdc80 and xNuf2. Using both RNAi and antibody injection experiments, we have extended previous characterization of the complex and found that Spc24 and Spc25 are required not only to establish, but also to maintain kinetochore-microtubule attachments and metaphase alignment. In addition, we show that Spc24 and Spc25 are required for chromosomal movement to the spindle poles in anaphase. PMID- 14738737 TI - The actin-interacting protein AIP1 is essential for actin organization and plant development. AB - Cell division, growth, and cytoplasmic organization require a dynamic actin cytoskeleton. The filamentous actin (F-actin) network is regulated by actin binding proteins that modulate actin dynamics. These actin binding proteins often have cooperative interactions. In particular, actin interacting protein 1 (AIP1) is capable of capping F-actin and enhancing the activity of the small actin modulating protein, actin depolymerising factor (ADF) in vitro. Here, we analyze the effect of the inducible expression of AIP1 RNAi in Arabidopsis plants to assess AIP1s role in vivo. In intercalary growing cells, the normal actin organization is disrupted, and thick bundles of actin appear in the cytoplasm. Moreover, in root hairs, there is the unusual appearance of actin cables ramifying the root hair tip. We suggest that the reduction in AIP1 results in a decrease in F-actin turnover and the promotion of actin bundling. This distortion of the actin cytoskeleton causes severe plant developmental abnormalities. After induction of the Arabidopis RNAi lines, the cells in the leaves, roots, and shoots fail to expand normally, and in the severest phenotypes, the plants die. Our data suggest that AIP1 is essential for the normal functioning of the actin cytoskeleton in plant development. PMID- 14738736 TI - Abstrakt, a DEAD box protein, regulates Insc levels and asymmetric division of neural and mesodermal progenitors. AB - Asymmetric cell division generates cell diversity in bacteria, yeast, and higher eukaryotes. In Drosophila, both neural and muscle progenitors divide asymmetrically. In these cells the Inscuteable (Insc) protein complex coordinates cell polarity and spindle orientation. Abstrakt (Abs) is a DEAD-box protein that regulates aspects of cell polarity in oocytes and embryos. We use a conditional allele of abs to investigate its role in neural and muscle progenitor cell polarity. In neuroblasts we observe loss of apical Insc crescents, failure in basal protein targeting, and defects in spindle orientation. In the GMC4-2a cell we observe loss of apical Insc crescents, defects in basal protein targeting, and equalization of sibling neuron fates; muscle precursors show a similar equalization of sibling cell fates. These phenotypes resemble those of insc mutants; indeed, abs mutants show a striking loss of Insc protein levels but no change of insc RNA levels. Furthermore, we find that the Abs protein physically interacts with insc RNA. Our results demonstrate a novel role for Abs in the posttranscriptional regulation of insc expression, which is essential for proper cell polarity, spindle orientation, and the establishment of distinct sibling cell fates within embryonic neural and muscle progenitors. PMID- 14738738 TI - Selection on codon usage in Drosophila americana. AB - Synonymous codons are not used at random, significantly influencing the base composition of the genome. The selection-mutation-drift model proposes that this bias reflects natural selection in favor of a subset of preferred codons. Previous estimates in Drosophila of the intensity of selective forces involved seem too large to be reconciled with theoretical predictions of the level of codon bias. This probably results from confounding effects of the demographic histories of the species concerned. We have studied three species of the virilis group of Drosophila, which are more likely to satisfy the assumptions of the evolutionary models. We analyzed the patterns of polymorphism and divergence in a sample of 18 genes and applied a new method for estimating the intensity of selection on synonymous mutations based on the frequencies of unpreferred mutations among polymorphic sites. This yielded estimates of selection intensities (N(e)s) of the order of 0.65, which is more compatible with the observed levels of codon bias. Our results support the action of both selection and mutational bias on codon usage bias and suggest that codon usage and genome base composition in the D. americana lineage are in approximate equilibrium. Biased gene conversion may also contribute to the observed patterns. PMID- 14738739 TI - Polarized light helps monarch butterflies navigate. AB - During their spectacular migratory journey in the fall, North American monarch butterflies (Danaus plexippus) use a time-compensated sun compass to help them navigate to their overwintering sites in central Mexico. One feature of the sun compass mechanism not fully explored in monarchs is the sunlight-dependent parameters used to navigate. We now provide data suggesting that the angle of polarized skylight (the e-vector) is a relevant orientation parameter. By placing butterflies in a flight simulator outdoors and using a linear polarizing filter, we show that manipulating the e-vector alters predictably the direction of oriented flight. Butterflies studied in either the morning or afternoon showed similar responses to filter rotation. Monarch butterflies possess the anatomical structure needed for polarized skylight detection, as rhabdoms in the dorsalmost row of photoreceptor cells in monarch eye show the organization characteristic of polarized-light receptors. The existence of polarized-light detection could allow migrants to accurately navigate under a variety of atmospheric conditions and reveals a critical input pathway into the sun compass mechanism. PMID- 14738740 TI - A noncoding RNA is required for the repression of RNApolII-dependent transcription in primordial germ cells. AB - RNApolII-dependent transcription is repressed in primordial germ cells of many animals during early development and is thought to be important for maintenance of germline fate by preventing somatic differentiation. Germ cell transcriptional repression occurs concurrently with inhibition of phosphorylation in the carboxy terminal domain (CTD) of RNApolII, as well as with chromatin remodeling. The precise mechanisms involved are unknown. Here, we present evidence that a noncoding RNA transcribed by the gene polar granule component (pgc) regulates transcriptional repression in Drosophila germ cells. Germ cells lacking pgc RNA express genes important for differentiation of nearby somatic cells and show premature phosphorylation of RNApolII. We further show that germ cells lacking pgc show increased levels of K4, but not K9 histone H3 methylation, and that the chromatin remodeling Swi/Snf complex is required for a second stage in germ cell transcriptional repression. We propose that a noncoding RNA controls transcription in early germ cells by blocking the transition from preinitiation to transcriptional elongation. We further show that repression of somatic differentiation signals mediated by the Torso receptor-tyrosine kinase is important for germline development. PMID- 14738741 TI - The radial positioning of chromatin is not inherited through mitosis but is established de novo in early G1. AB - The organization of chromatin in the nucleus is nonrandom. Different genomic regions tend to reside in preferred nuclear locations, relative to radial position and nuclear compartments. Several lines of evidence support a role for chromatin localization in the regulation of gene expression. Therefore, a key problem is how the organization of chromatin is established and maintained in dividing cell populations. There is controversy about the extent to which chromatin organization is inherited from mother to daughter nucleus. We have used time-lapse microscopy to track specific human loci after exit from mitosis. In comparison to later stages of interphase, we detect increased chromatin mobility during the first 2 hr of G1, and during this period association of loci with nuclear compartments is both gained and lost. Although chromatin in daughter nuclei has a rough symmetry in its spatial distribution, we show, for the first time, that the association of loci with nuclear compartments displays significant asymmetry between daughter nuclei and therefore cannot be inherited from the mother nucleus. We conclude that the organization of chromatin in the nucleus is not passed down precisely from one cell to its descendents but is more plastic and becomes refined during early G1. PMID- 14738742 TI - Business pressure mounts. PMID- 14738743 TI - A degree of profit. PMID- 14738744 TI - Q & A. PMID- 14738745 TI - Systems biology versus molecular biology. PMID- 14738746 TI - ELAV. PMID- 14738748 TI - Recombination: Holliday junction resolution and crossover formation. AB - The heterodimeric nuclease Mus81-Eme1 has been proposed to be a Holliday junction resolvase and has now been found to be responsible for nearly all meiotic crossovers in fission yeast. The intriguing substrate preference of this enzyme for nicked Holliday junctions opens the possibility that crossover formation may not always involve double Holliday junctions. PMID- 14738747 TI - Linking transcriptional mediators via the GACKIX domain super family. PMID- 14738749 TI - Protein degradation: CUL-3 and BTB--partners in proteolysis. AB - In early C. elegans embryos, the transition from meiosis to mitosis requires degradation of the MEI-1 protein. A novel class of SCF-like ubiquitin ligases has been identified that mediates this process. These ligases contain the CUL-3 scaffold at their core and use a BTB-domain protein in substrate recognition. PMID- 14738750 TI - Neuronal development: specifying a hard-wired circuit. AB - The formation of neuronal circuits that relay distinct olfactory information is thought to depend on cues provided by pre-synaptic receptor neurons. But direct visualization of second order neurons in Drosophila now suggests that dendritic targeting occurs independently of interactions with incoming sensory neurons. PMID- 14738751 TI - Microbial genomics: tropical treasure? AB - A Brazilian consortium has unveiled the genomic DNA sequence of the purple pigmented bacterium Chromobacterium violaceum, a dominant component of the tropical soil microbiota. The sequence provides insight into the abundant potential of this organism for biotechnological and pharmaceutical applications. PMID- 14738752 TI - Hedgehog signaling: Costal-2 bridges the transduction gap. AB - Drosophila genetics has provided the Hedgehog signaling pathway with roughly ordered components, but few mechanistic connections. Several studies now show that the kinesin-related protein Costal-2 provides a physical link between the transmembrane transducer Smoothened and the transcriptional effector Cubitus interruptus. PMID- 14738753 TI - Cell division: AAAtacking the mitotic spindle. AB - Cell division requires the assembly of a microtubule-based spindle which captures and segregates sister chromatids. But how is this spindle broken down once chromosome segregation is complete? New evidence implicates a highly conserved AAA-ATPase in spindle disassembly at the end of mitosis. PMID- 14738754 TI - Evolution of cooperation: two for one? AB - How can cooperation thrive in a selfish world? Recent evolution experiments show how bacteria themselves can generate conditions that make cooperation a winning strategy. At least in the short term. PMID- 14738755 TI - Genomes: a helpful cousin for our favourite worm. AB - The recently published genome of the nematode Caenorhabditis briggsae provides a drastic improvement in structural annotation of the C. elegans genome, as well as a promising source of evolutionary comparisons. PMID- 14738756 TI - Molecular chaperones: structure of a protein disaggregase. AB - The ring-forming molecular chaperone Hsp104/ClpB is a member of the AAA+ protein family which rescues proteins from aggregated states. The newly determined crystal structure of ClpB provides new insights into the mechanism of protein disaggregation, suggesting a crowbar activity mediated by a unique coiled-coil domain. PMID- 14738757 TI - Something in the air? New insights into mammalian pheromones. AB - Olfaction is the dominant sensory modality for most animals and chemosensory communication is particularly well developed in many mammals. Our understanding of this form of communication has grown rapidly over the last ten years since the identification of the first olfactory receptor genes. The subsequent cloning of genes for rodent vomeronasal receptors, which are important in pheromone detection, has revealed an unexpected diversity of around 250 receptors belonging to two structurally different classes. This review will focus on the chemical nature of mammalian pheromones and the complementary roles of the main olfactory system and vomeronasal system in mediating pheromonal responses. Recent studies using genetically modified mice and electrophysiological recordings have highlighted the complexities of chemosensory communication via the vomeronasal system and the role of this system in handling information about sex and genetic identity. Although the vomeronasal organ is often regarded as only a pheromone detector, evidence is emerging that suggests it might respond to a much broader variety of chemosignals. PMID- 14738758 TI - "Homing to Niche," a new criterion for hematopoietic stem cells? AB - By combining cell surface staining with fluorochrome-conjugated monoclonal antibodies and Hoechst 33342 dye supravital staining, Matsuzaki et al. have succeeded in enriching hematopoietic stem cells (HSCs) essentially to homogeneity. When single-cell transplantation analysis was performed using the isolated cells, over 95% of the recipient mice showed long-term multilineage engraftment. The work demonstrates unexpectedly high marrow seeding efficiency of HSCs and proposes high marrow homing capacity as a new criterion for HSCs. PMID- 14738759 TI - IL-6 trans-signaling: the heat is on. AB - The molecular consequence of the fever response has been illuminated by a recent study showing that a temperature shift to 40 degrees C resulted in increased leukocyte adhesion to tissue sections, which was mediated by L-selectin activation in lymphocytes. This L-selectin activation during heat responses was dependent on IL-6 trans-signaling via the soluble IL-6R. PMID- 14738760 TI - Plasticity of T cell memory responses to viruses. AB - Virus-specific memory T cell populations demonstrate plasticity in antigenic and functional phenotype, in recognition of antigen, and in their ability to accommodate new memory T cell populations. The adaptability of complex antigen specific T cell repertoires allows the host to respond to a diverse array of pathogens and accommodate memory pools to many pathogens in a finite immune system. This is in part accounted for by crossreactive memory T cells, which can be employed in immune responses and mediate protective immunity or life threatening immunopathology. PMID- 14738761 TI - Dendritic cells: immunobiology and cancer immunotherapy. AB - A recent workshop on "Dendritic Cells: Biology and Therapeutic Applications," sponsored by the Juan March Foundation, brought together basic and clinical research scientists to discuss the mechanisms underlying the control of immune responses and tolerance by dendritic cells (DCs), as well as recent research in cancer immunotherapy based on DC vaccination. Particular emphasis was placed on antigen processing and presentation by DCs, C-type lectin antigen receptors, DC maturation and polarization of T cell responses, the control of immunity versus tolerance by DCs, the developmental origin of DCs, and the use of DCs in cancer immunotherapy. PMID- 14738762 TI - RhoA and zeta PKC control distinct modalities of LFA-1 activation by chemokines: critical role of LFA-1 affinity triggering in lymphocyte in vivo homing. AB - Chemokines regulate rapid leukocyte adhesion by triggering a complex modality of integrin activation. We show that the small GTPase RhoA and the atypical zeta PKC differently control lymphocyte LFA-1 high-affinity state and rapid lateral mobility induced by chemokines. Activation of LFA-1 high-affinity state and lateral mobility is controlled by RhoA through the activity of distinct effector regions, demonstrating that RhoA is a central point of diversification of signaling pathways leading to both modalities of LFA-1 triggering. In contrast, zeta PKC controls LFA-1 lateral mobility but not affinity triggering. Blockade of the 23-40 RhoA effector region prevents induction of LFA-1 high-affinity state as well as lymphocyte arrest in Peyer's patch high endothelial venules. Thus, RhoA controls the induction of LFA-1 high-affinity state by chemokines independently of zeta PKC, and this is critical to support chemokine-regulated homing of circulating lymphocytes. PMID- 14738764 TI - Dendritic cells transfected with cytopathic self-replicating RNA induce crosspriming of CD8+ T cells and antiviral immunity. AB - A potential shortcoming of nonlive vaccines is their relative inefficiency in generating T cell responses, thus limiting their application in infections requiring cellular immunity. Here, we present a system to induce cellular immunity and to study the immunological implications of time-delayed dendritic cell (DC) apoptosis and antigen reprocessing in vivo. We generated a self replicating cytopathic pestivirus RNA to enhance production and presentation of hepatitis C virus (HCV) antigens and to induce apoptosis in DC 24-48 hr after transfection. Replicon-transfected H-2(b) DCs used to immunize HLA-A2 transgenic mice induced protection upon challenge with a vaccinia virus expressing HCV antigens. Induction of cell death enhanced the immunogenicity of DC-associated antigen. Transfer of cellular material from vaccine DCs to endogenous antigen presenting cells was visualized in lymph nodes and spleen, and crossprimed CD8(+) T cells were characterized. The findings are relevant for the rational design of vaccines against noncytopathic pathogens like HCV. PMID- 14738763 TI - Regulation of ZAP-70 activation and TCR signaling by two related proteins, Sts-1 and Sts-2. AB - T cells play a central role in the recognition and elimination of foreign pathogens. Signals through the T cell receptor (TCR) control the extent and duration of the T cell response. To ensure that T cells are not inappropriately activated, signaling pathways downstream of the TCR are subject to multiple levels of positive and negative regulation. Herein, we describe two related proteins, Sts-1 and Sts-2, that negatively regulate TCR signaling. T cells from mice lacking Sts-1 and Sts-2 are hyperresponsive to TCR stimulation. The phenotype is accompanied by increased Zap-70 phosphorylation and activation, including its ubiquitinylated forms. Additionally, hyperactivation of signaling proteins downstream of the TCR, a marked increase in cytokine production by Sts1/2(-/-) T cells, and increased susceptibility to autoimmunity in a mouse model of multiple sclerosis is observed. Therefore, Sts-1 and Sts-2 are critical regulators of the signaling pathways that regulate T cell activation. PMID- 14738765 TI - Central role of IL-6 receptor signal-transducing chain gp130 in activation of L selectin adhesion by fever-range thermal stress. AB - The physiological benefit of the febrile response is poorly understood. Here we show that fever-range thermal stress enhances the function of the L-selectin lymphocyte homing receptor through an interleukin-6 (IL-6)-dependent signaling mechanism. Thermal stimulation of L-selectin adhesion in vitro and in vivo is mediated by engagement of the gp130 signal-transducing chain by IL-6 and a soluble form of the IL-6 receptor-alpha (sIL-6Ralpha) binding subunit. Thermal control of adhesion is maintained in IL-6-deficient mice through a gp130 dependent compensatory mechanism mediated by IL-6-related cytokines (i.e., oncostatin M [OSM], leukemia inhibitory factor [LIF], and IL-11). Combined biochemical and pharmacological inhibitor (PD98059, U0126, SB203580, SP600125) approaches positioned MEK1/ERK1-2, but not p38 MAPK or JNK, in the IL-6/sIL 6Ralpha signaling pathway upstream of activation of L-selectin/cytoskeletal interactions and L-selectin avidity/affinity. These results highlight a role for gp130-linked IL-6/sIL-6Ralpha transsignaling in amplifying lymphocyte trafficking during febrile inflammatory responses. PMID- 14738766 TI - Human cytomegalovirus inhibits tapasin-dependent peptide loading and optimization of the MHC class I peptide cargo for immune evasion. AB - The immune evasion protein US3 of human cytomegalovirus binds to and arrests MHC class I molecules in the endoplasmic reticulum (ER). However, substantial amounts of class I molecules still escape US3-mediated ER retention, suggesting that not all class I alleles are affected equally by US3. Here, we identify tapasin inhibition as the mechanism of MHC retention by US3. US3 directly binds tapasin and inhibits tapasin-dependent peptide loading, thereby preventing the optimization of the peptide repertoire presented by class I molecules. Due to the allelic specificity of tapasin toward class I molecules, US3 affects only class I alleles that are dependent on tapasin for peptide loading and surface expression. Accordingly, tapasin-independent class I alleles selectively escape to the cell surface. PMID- 14738767 TI - Unexpectedly efficient homing capacity of purified murine hematopoietic stem cells. AB - Single-cell transplantation analysis revealed that the cells that had the strongest dye efflux activity ("Tip"-SP cells) and had the phenotype CD34- c-Kit+ Sca-1+ Lin- (CD34- KSL cells) exhibited very strong proliferation and multilineage differentiation capacity. Ninety-six percent of the lethally irradiated mice that received a single "Tip"-SP CD34- KSL cell showed significant donor cell engraftment for long term. These findings support the hypothesis that "Tip"-SP CD34- KSL cells represent the most primitive hematopoietic stem cells that are capable of migrating into the primary site and surviving and/or proliferating with nearly absolute efficiency. This led us to propose high marrow seeding efficiency as a specific characteristic of primitive HSCs, in addition to their self-renewal and multipotent capacity. PMID- 14738768 TI - HSP70 peptide binding mutants separate antigen delivery from dendritic cell stimulation. AB - Microbial heat shock proteins (HSPs) have been implicated in the induction of both the innate and adaptive arms of the immune response. We now show that human dendritic cells (DC) pulsed with peptide-loaded mycobacterial HSP70 complexes generate potent antigen-specific cytotoxic lymphocyte (CTL) responses, which are dependent on an HSP70-stimulated calcium signaling cascade. From the calculated peptide binding affinity of mycobacterial HSP70 (K(D) = 14 microM) we show that 120 pM HSP70 bound peptide is sufficient to generate a peptide-specific CTL response that is up to four orders of magnitude more efficient than peptide alone. The minimal 136 amino acid, mycobacterial HSP70 peptide binding domain can generate CTL responses, and a single amino acid mutant HSP70 designed to prevent peptide binding but retain stimulatory capacity has allowed us to separate antigen delivery from DC immunostimulation. PMID- 14738769 TI - Tumor-specific human CD4+ regulatory T cells and their ligands: implications for immunotherapy. AB - Regulatory T cells play an important role in the maintenance of immunological self-tolerance by suppressing immune responses against autoimmune diseases and cancer. Little is known, however, about the nature of the physiological target antigens for CD4(+) regulatory T (Treg) cells. Here we report the identification of the LAGE1 protein as a ligand for tumor-specific CD4(+) Treg cell clones generated from the tumor-infiltrating lymphocytes (TILs) of cancer patients. Phenotypic and functional analyses demonstrated that they were antigen-specific CD4(+) Treg cells expressing CD25 and GITR molecules and possessing suppressive activity on the proliferative response of naive CD4(+) T cells to anti-CD3 antibody stimulation. Ligand-specific activation and cell-cell contact were required for TIL102 Treg cells to exert suppressive activity on CD4(+) effector cells. These findings suggest that the presence of tumor-specific CD4(+) Treg cells at tumor sites may have a profound effect on the inhibition of T cell responses against cancer. PMID- 14738770 TI - Category coherence and category-based property induction. AB - One important property of human object categories is that they define the sets of exemplars to which newly observed properties are generalized. We manipulated the causal knowledge associated with novel categories and assessed the resulting strength of property inductions. We found that the theoretical coherence afforded to a category by inter-feature causal relationships strengthened inductive projections. However, this effect depended on the degree to which the exemplar with the to-be-projected predicate manifested or satisfied its category's causal laws. That is, the coherence that supports inductive generalizations is a property of individual category members rather than categories. Moreover, we found that an exemplar's coherence was mediated by its degree of category membership. These results were obtained across a variety of causal network topologies and kinds of categories, including biological kinds, non-living natural kinds, and artifacts. PMID- 14738771 TI - A unique look at face processing: the impact of masked faces on the processing of facial features. AB - This experiment utilized a masked priming paradigm to explore the early processes involved in face recognition. The first experiment investigated implicit processing of the eyes and mouth in an upright face, using prime durations of 33 and 50 ms. The results demonstrate implicit processing of both the eyes and mouth, and support the configural processing theory of face processing. The second experiment used the same method with inverted faces and the third experiment was a combination of Experiments 1 and 2. The fourth experiment utilized misaligned faces as the primes. Based on the pattern of results from these experiments, we suggest that, when a face is inverted, the eyes and mouth are initially processed individually and are not linked until a later stage of processing. An upright face is proposed to be processed by analysis of its configuration, whereas an inverted face is initially processed using first-order relational information, and then converted to an upright representation and transferred to face specific regions for configural analysis. PMID- 14738772 TI - Infants chunk object arrays into sets of individuals. AB - Research suggests that, using representations from object-based attention, infants can represent only 3 individuals at a time. For example, infants successfully represent 1, 2, or 3 hidden objects, but fail with 4 (Developmental Science 6 (2003) 568), and a similar limit is seen in adults' tracking of multiple objects (see Cognitive Psychology 38 (1999) 259). In the present experiments we used a manual search procedure to ask whether infants can overcome this limit of 3 by chunking individuals into sets. Experiments 1 and 2 replicate infants' failure to represent a total of 4 objects. We then show that infants can exceed this limit when items are spatiotemporally grouped into two sets of 2 prior to hiding, leading infants to successfully represent a total of 4 objects. Experiment 3 demonstrates that infants tracked the 4 objects as two sets of 2, searching for each set in its correct hiding location. That infants represented the number of individuals in each set is demonstrated by their reaching for the correct number of objects in each location. These results suggest that by binding individuals into sets, infants can increase their representational capacity. This is the first evidence for chunking abilities in infants. PMID- 14738773 TI - Commanding the direction of passive whole-body rotations facilitates egocentric spatial updating. AB - In conditions of slow passive transport without vision, even tenuous inertial signals from semi-circular canals and the haptic-kinaesthetic system should provide information about changes relative to the environment provided that it is possible to command the direction of the body's movements voluntarily. Without such control, spatial updating should be impaired because incoming signals cannot be compared to the expected sensory consequences provided by voluntary command. Participants were seated in a rotative robot (Robuter) and learnt the positions of five objects in their surroundings. They were then blindfolded and assigned either to the active group (n=7) or to the passive group (n=7). Members of the active group used a joystick to control the direction of rotation of the robot. The acceleration (25 degrees /s2) and plateau velocity (9 degrees /s) were kept constant. The participants of the passive group experienced the same stimuli passively. After the rotations, the participants had to point to the objects whilst blindfolded. Participants in the active group significantly outperformed the participants in the passive group. Thus, even tenuous inertial cues are useful for spatial updating in the absence of vision, provided that such signals are integrated as feedback associated with intended motor command. PMID- 14738774 TI - Lexical orthographic knowledge develops from the beginning of literacy acquisition. AB - This study reports two experiments assessing the spelling performance of French first graders after 3 months and after 9 months of literacy instruction. The participants were asked to spell high and low frequency irregular words (Experiment 1) and pseudowords, some of which had lexical neighbours (Experiment 2). The lexical database which children had been exposed to was strictly controlled. Both a frequency effect in word spelling accuracy and an analogy effect in pseudoword spelling were obtained after only 3 months of reading instruction. The results suggest that children establish specific orthographic knowledge from the very beginning of literacy acquisition. PMID- 14738775 TI - Baby arithmetic: one object plus one tone. AB - Recent studies using a violation-of-expectation task suggest that preverbal infants are capable of recognizing basic arithmetical operations involving visual objects. There is still debate, however, over whether their performance is based on any expectation of the arithmetical operations, or on a general perceptual tendency to prefer visually familiar and complex displays. Here we provide new evidence that 5-month-old infants recognize basic arithmetic operations across sensory modalities. Using a violation-of-expectation task that eliminated the possibility of the familiarity and complexity preference, 5-month-old infants were presented alternatively with two types of arithmetical events: the expected, correct outcomes of operations (1 object+1 tone=2 objects and 1 object+2 tones=3 objects) and the unexpected, incorrect ones (1 object+2 tones=2 objects and 1 object+1 tone=3 objects). Results showed that subjects looked significantly longer at the unexpected events than at the expected events, suggesting that infants are able to recognize basic arithmetic operations across sensory modalities. PMID- 14738776 TI - Development of a monoclonal antibody based competitive-ELISA for detection and titration of antibodies to peste des petits ruminants (PPR) virus. AB - Peste des petits ruminants (PPR) is an acute febrile, viral, disease of small ruminants with great economic importance. A competitive-ELISA (c-ELISA) test was developed for detection of antibodies to PPR virus in the sera samples of goats and sheep. The test uses monoclonal antibody to a neutralizing epitope of haemagglutinin protein of the virus. Based on the distribution of known negative sera samples (n=933) in respect of PPR virus antibodies in the test, a cut-off value was set as 38%. This value was the result of mean of negative population added with two times the standard deviations. A total of 1668 sera samples from goat and sheep and 32 sera from cattle were screened by c-ELISA and virus neutralization test (VNT). Efficacy of c-ELISA compared very well with VNT having high relative specificity (98.4%) and sensitivity (92.4%). The sensitivity of c ELISA for PPR sero-surveillance could further be increased (95.4%), if the target population is non-vaccinated. c-ELISA test correlated well with VNT (r=0.845) for end-point titration of PPR virus antibody in 64 goat sera samples. It could clearly separate infected population from uninfected in field sera. Using c-ELISA test paired sera samples from 13 goats provided a clear diagnosis of PPR virus infection. Furthermore, antibodies to PPR virus could be successfully detected during 1 year after vaccination in four goats inoculated with an experimental PPR vaccine. Findings suggest that the c-ELISA test developed can easily replace VNT for sero-surveillance, sero-monitoring, diagnosis from paired sera samples and end-point titration of PPR virus antibodies. PMID- 14738777 TI - Evaluation of molecular and immunological techniques for the diagnosis of mammary aspergillosis in ewes. AB - Three techniques for the diagnosis of mammary aspergillosis in ewes were compared: indirect ELISA to detect the level of anti-Aspergillus IgG in serum, determination of galactomannan (Platelia procedure), and detection of DNA of Aspergillus in serum by a nested PCR. Twenty sera from proven cases of aspergillosis in ewes were positive using ELISA (100%), 80% were positive using PCR, but only 55% were positive using Platelia. All 20 control sera were negative using ELISA and PCR, whereas using Platelia methodology one was positive and the other doubtful. The detection of antibody by ELISA in sera is therefore a reliable criterion for the diagnosis of mammary aspergillosis in ewes. Platelia showed the same deficiencies reported in humans, with the appearance of false positives and negatives. The use of PCR was promising and might have valuable application in human medicine. PMID- 14738778 TI - Transmission of multiple antimicrobial-resistant Staphylococcus intermedius between dogs affected by deep pyoderma and their owners. AB - The occurrence of antimicrobial-resistant Staphylococcus intermedius strains was investigated in 13 dogs affected by deep pyoderma, their owners and 13 individuals without daily contact with dogs (control group). A total of 90 canine and 33 human S. intermedius isolates were typed by pulsed field gel electrophoresis (PFGE) to determine their possible identity. The occurrence of S. intermedius in dog-owners was significantly higher compared with the control group (Fisher's exact test, P=0.03), with S. intermedius being detected in seven dog-owners and in one individual not exposed to dogs. The results of the PFGE analysis showed that six out of 13 (46%) owners carried strains identical to those isolated from their dogs. Strains detected in both dogs and humans were resistant up to five different antimicrobial classes, including penicillins, fusidic acid, macrolides/lincosamides, tetracycline and chloramphenicol. Based on the results of this study, owners of dogs affected by deep pyoderma often carry multiple antimicrobial-resistant strains of S. intermedius occurring in their dogs. Independent of the direction and modalities of transmission, this finding raises questions concerning the possible transfer of resistance genes from canine S. intermedius to human pathogenic staphylococci. PMID- 14738779 TI - No foot-and-mouth disease virus transmission between individually housed calves. AB - The foot-and-mouth disease outbreak in The Netherlands in 2001 most likely started on a mixed veal-calf/dairy-goat farm. The outbreak among the 74 calves on this farm appeared to be limited to four animals, and no clinical signs of FMD were reported. Also on a second veal-calf farm minor clinical signs and limited virus transmission were observed. Since FMD is known to be a very contagious disease, and can cause severe lesions, these observations were disputed. Therefore, we carried out two experiments to determine whether the Dutch FMD virus isolate from 2001 does spread among individually housed calves with limited contacts, either indirect (experiment 1) or direct (experiment 2). In experiment 1, four pairs of calves were housed in an individual box at 1m distance from each other. In experiment 2, two groups of three calves were housed in individual boxes, directly bordering each other. We infected one animal per pair in experiment 1, and the calf in the middle in experiment 2. We recorded clinical signs, virus shedding in saliva and the development of antibodies. In addition, we determined whether the virus was transmitted from the inoculated calves to the neighbour(s). All inoculated calves showed mild signs of FMD--fever, and some vesicles on hooves and/or in the mouth--but only one calf showed signs that were visible without physical examination. All inoculated calves shed virus in the saliva and developed neutralising antibodies. None of the contact animals seroconverted, indicating that virus transmission did not occur. These experiments showed that no virus transmission among individual housed calves can occur. This finding supports the hypothesis of the route of virus introduction to The Netherlands in 2001 and show that the observations on the two veal-calf farms were not impossible. PMID- 14738780 TI - Intestinal colonisation-inhibition and virulence of Salmonella phoP, rpoS and ompC deletion mutants in chickens. AB - Administration of live Salmonella strains to day-old chicks provides profound protection against superinfection with a related strain within a matter of hours by a colonisation-inhibition mechanism, which is primarily a bacterial physiological process. Although currently available, commercial, live attenuated Salmonella vaccines induce protection by adaptive immunity, none of them is able to induce protection against Salmonella organisms by colonisation-inhibition and, therefore, they are unable to protect newly-hatched birds immediately after oral vaccination. In this study, mutants of Salmonella Typhimurium and Enteritidis with deletions in phoP and rpoS, either alone or in combination with ompC, were characterised and tested for their level of attenuation and their ability to inhibit the intestinal colonisation of the isogenic parent strains in chickens. Mutants with deletions only in rpoS demonstrated an unaffected potential to inhibit the intestinal colonisation of the challenge strain but were still fully virulent for the chickens. Mutants with deletions in phoP, either alone or in combination with rpoS, resulted in a high level of attenuation, unimpaired ability to colonise the gut and a nearly unaffected potential to inhibit the challenge strain from caecal colonisation. Mutants with an additional deletion in ompC revealed a reduced capacity of intestinal colonisation-inhibition when compared to the control strains and both the single rpoS and the phoP deletion mutants. Mutations in phoP- or phoP-regulated genes may therefore be used for the development of live attenuated Salmonella vaccines possessing these novel characteristics. PMID- 14738781 TI - Influence of porcine intestinal pH and gastric digestion on antigenicity of F4 fimbriae for oral immunisation. AB - Newly weaned piglets can be orally immunised against F4+ enterotoxigenic Escherichia coli (ETEC) infection with F4 fimbriae. However, to efficiently develop a vaccine against ETEC induced postweaning diarrhoea, knowledge of the stability of the F4 fimbriae to different pH and gastric digestion is needed. The gastrointestinal pH in suckling and recently weaned piglets was measured and the stability of F4 fimbriae to different pH and to pepsin was assessed in vitro. In the stomach the lowest pH was found in the fundus gland region. Gastric pH values below 2.5 were not found in suckling piglets or at the day of weaning, in contrast to piglets 1 and 2 weeks postweaning. Along the first half of the small intestine and in the caecum, a negative correlation was found between pH and age. The F4 fimbriae were stable to pH 1.5 and 2 for 2 h, whereas longer incubation periods resulted in conversion of the multimeric forms into monomers. The F4 fimbriae were partially degraded by incubation for 15-30 min in simulated gastric fluid at pH 1.5 and 2, and completely digested from 3 h onwards. At pH 3, the fimbriae maintained their antigenicity for at least 4h. The results demonstrate that gastric digestion will only have a limited impact on oral immunisation since liquid passes through the stomach relatively quickly (50% within 2 h). However, we previously demonstrated that the transit times are prolonged shortly after weaning. Shortly after weaning it could be necessary to protect the F4 fimbriae against gastric digestion to obtain efficient oral immunisation of the piglets. PMID- 14738782 TI - Antimicrobial resistance of commensal Escherichia coli from dairy cattle associated with recent multi-resistant salmonellosis outbreaks. AB - The use of antimicrobial drugs in livestock is suspected to contribute to bacterial antimicrobial resistance (AR) development. Dairy farms experiencing recent outbreaks of salmonellosis involving multi-resistant (MR) Salmonella strains were compared to control farms with respect to AR among bovine commensal E. coli isolates. For most antimicrobials tested, the percentage of AR E. coli isolated from salmonellosis-affected farms was significantly higher than that from control farms. Calf E. coli from both case and control farms had greater levels of AR than cow isolates. Commensal E. coli isolates from case farms and calves tended to more frequently be MR. These data are consistent with the existence of higher antimicrobial selection pressure on farms with recent salmonellosis outbreaks, however, the directionality of the relationship remains to be elucidated. An improved understanding of the epidemiology of AR bacteria in livestock production, both at the herd and molecular level, is essential to mitigate risk to public health and food safety. PMID- 14738783 TI - Identification of Bartonella strains isolated from wild and domestic ruminants by a single-step PCR analysis of the 16S-23S intergenic spacer region. AB - Of the 20 species or subspecies of Bartonella currently known, 7 cause various diseases in humans with many being zoonotic. However, some Bartonella species appear only to cause asymptomatic bacteraemia in their hosts. In ruminants, three Bartonella species (B. bovis, B. capreoli and B. schoenbuchensis) have recently been described. However, limited or no information has yet been published concerning their mode of transmission and their possible pathogenicity for domestic cattle. The phylogenetic relationship of these species with other bacteria of the Bartonella genus has only been recently investigated. It is therefore necessary to develop appropriate tools that will easily allow identification of these ruminant strains for epidemiological and clinical studies. A single-step PCR assay, based on the amplification of a fragment of the 16S-23S rRNA intergenic spacer (ITS), was evaluated for identification of Bartonella isolated from domestic cattle and from free-ranging or captive cervids. For each Bartonella species tested, the PCR assay led to a product that was unique either for its length or its sequence. All ruminant isolates tested could be easily differentiated among themselves and from the other Bartonella species. Furthermore, sequence analysis of the PCR products revealed a close relationship between all ruminant Bartonella strains. Therefore, ITS PCR testing appears to be a convenient tool for a quick diagnosis of ruminant Bartonella species. PMID- 14738784 TI - 2004 US election campaign: health at home and abroad. PMID- 14738785 TI - Schistosoma japonicum: modern tools for an ancient disease. PMID- 14738786 TI - Treatment of Guillain-Barre syndrome with corticosteroids: lack of benefit? PMID- 14738787 TI - Physical activity and obesity. PMID- 14738788 TI - The beta-agonist controversy revisited. PMID- 14738789 TI - Research and development costs for drugs. PMID- 14738790 TI - Sudden unexplained infant death in 20 regions in Europe: case control study. AB - BACKGROUND: After striking changes in rates of sudden unexplained infant death (SIDS) around 1990, four large case-control studies were set up to re-examine the epidemiology of this syndrome. The European Concerted Action on SIDS (ECAS) investigation was planned to bring together data from these and new studies to give an overview of risk factors for the syndrome in Europe. METHODS: We undertook case-control studies in 20 regions. Data for more than 60 variables were extracted from anonymised records of 745 SIDS cases and 2411 live controls. Logistic regression was used to calculate odds ratios (ORs) for every factor in isolation, and to construct multivariate models. FINDINGS: Principal risk factors were largely independent. Multivariately significant ORs showed little evidence of intercentre heterogeneity apart from four outliers, which were eliminated. Highly significant risks were associated with prone sleeping (OR 13.1 [95% CI 8.51-20.2]) and with turning from the side to the prone position (45.4 [23.4 87.9]). About 48% of cases were attributable to sleeping in the side or prone position. If the mother smoked, significant risks were associated with bed sharing, especially during the first weeks of life (at 2 weeks 27.0 [13.3-54.9]). This OR was partly attributable to mother's consumption of alcohol. Mother's alcohol consumption was significant only when baby bed-shared all night (OR increased by 1.66 [1.16-2.38] per drink). For mothers who did not smoke during pregnancy, OR for bed-sharing was very small (at 2 weeks 2.4 [1.2-4.6]) and only significant during the first 8 weeks of life. About 16% of cases were attributable to bed-sharing and roughly 36% to the baby sleeping in a separate room. INTERPRETATION: Avoidable risk factors such as those associated with inappropriate infants' sleeping position, type of bedding used, and sleeping arrangements strongly suggest a basis for further substantial reductions in SIDS incidence rates. PMID- 14738791 TI - Effect of methylprednisolone when added to standard treatment with intravenous immunoglobulin for Guillain-Barre syndrome: randomised trial. AB - BACKGROUND: Despite available treatment with intravenous immunoglobulin (IVIg), morbidity and mortality are considerable in patients with Guillain-Barre syndrome (GBS). Our aim was to assess whether methylprednisolone, when taken with IVIg, improves outcome when compared with IVIg alone. METHODS: We did a double-blind, placebo-controlled, multicentre, randomised study, to which we enrolled patients who were unable to walk independently and who had been treated within 14 days after onset of weakness with IVIg (0.4 g/kg bodyweight per day) for 5 days. We assigned 233 individuals to receive either intravenous methylprednisolone (500 mg per day; n=116) or placebo (n=117) for 5 days within 48 h of administration of first dose of IVIg. Because age is an important prognostic factor, we split treatment groups into two age-groups-ie, younger than age 50 years, or 50 years and older. Our primary outcome was an improvement from baseline in GBS disability score of one or more grades 4 weeks after randomisation. Analysis was by intention to treat. FINDINGS: We analysed 225 patients. GBS disability scores increased by one grade or more in 68% (76 of 112) of patients in the methylprednisolone group and in 56% (63 of 113) of controls (odds ratio [OR] 1.68, 95% CI 0.97-2.88; p=0.06). After adjustment for age and degree of disability at entry, treatment OR was 1.89 (95% CI 1.07-3.35; p=0.03). Side effects did not differ greatly between groups. INTERPRETATION: We noted no significant difference between treatment with methylprednisolone and IVIg and IVIg alone. Because of the relevance of prognostic factors and the limited side effects of methylprednisolone, the potential importance of combination treatment with the drug and IVIg, however, warrants further investigation. PMID- 14738792 TI - Prediction of total and hip fracture risk in men and women by quantitative ultrasound of the calcaneus: EPIC-Norfolk prospective population study. AB - BACKGROUND: A quarter of fractures needing admission happen in men, but few data are available that show the value of bone measures for prediction of fracture risk in men. We aimed to assess quantitative ultrasound of the calcaneum and fracture incidence in a prospective observational population study. METHODS: Calcaneum broadband ultrasound attenuation (BUA) was measured in men and women in the Norfolk cohort of the European Prospective Investigation into Cancer (EPIC Norfolk) between 1997 and 2000. Incident fractures were ascertained by hospital record linkage. FINDINGS: In 14824 men and women aged 42-82 years, during mean follow-up of 1.9 years (SD 0.7), there were 121 incident fractures that needed admission, including 31 hip fractures. Men and women in the lowest 10% of the calcaneum BUA distribution had a relative risk of fracture of 4.44 (95% CI 2.24 8.89, p<0.0001) compared with those in the upper 30% of the distribution. A fall of about 1 SD in BUA (20 db/MHz) was associated with a relative risk of fracture of 1.95 (95% CI 1.50-2.52, p<0.0001), independent of age, sex, weight, height, cigarette smoking habit, and past history of fracture. BUA predicted fractures with consistent magnitude in subgroups stratified by sex, age 65 years or older and younger than 65 years, smoking habit, past history of fracture, and hip and non-hip fractures separately. The sex difference in fracture risk was largely accounted for by differences in BUA. INTERPRETATION: Quantitative calcaneum ultrasound predicts total and hip fracture risk in men and women in a continuous relation. The challenge now is to identify interventions to improve bone health in the whole population. PMID- 14738793 TI - Role of interleukin 6 in myocardial dysfunction of meningococcal septic shock. AB - BACKGROUND: Myocardial failure has a central role in the complex pathophysiology of septic shock and contributes to organ failure and death. During the sepsis induced inflammatory process, specific factors are released that depress myocardial contractile function. We aimed to identify these mediators of myocardial depression in meningococcal septic shock. METHODS: We combined gene expression profiling with protein and cellular methods to identify a serum factor causing cardiac dysfunction in meningococcal septic shock. We identified genes that were significantly upregulated in blood after exposure to meningococci. We then selected for further analysis those genes whose protein products had properties of a myocardial depressant factor--specifically a 12-25 kDa heat stable protein that is released into serum shortly after onset of meningococcal infection. FINDINGS: We identified 174 significantly upregulated genes in meningococcus-infected blood: six encoded proteins that were of the predicted size and had characteristics of a myocardial depressant factor. Of these, interleukin 6 caused significant myocardial depression in vitro. Removal of interleukin 6 from serum samples of patients with meningococcaemia and from supernatants of inflammatory cells stimulated by meningococci in vitro abolished the negative inotropic activity. Furthermore, concentrations in serum of interleukin 6 strongly predicted degree of myocardial dysfunction and severity of disease in children with meningococcal septic shock. INTERPRETATION: Interleukin 6 is a mediator of myocardial depression in meningococcal disease. This cytokine and its downstream mediators could be a target for future treatment strategies. PMID- 14738794 TI - Congestive heart failure and a swollen breast. PMID- 14738795 TI - Total energy expenditure and physical activity in young Scottish children: mixed longitudinal study. AB - Childhood obesity has been attributed to a decline in total energy expenditure (TEE). We measured TEE, physical activity, and sedentary behaviour in a representative sample of young children from Glasgow, UK, at age 3 years (n=78), and we did a follow-up study at age 5 years (n=72). Mean physical activity level (TEE/resting energy expenditure) was 1.56 (SD 0.39) at age 3 years and 1.61 (0.22) at age 5 years. Median time in sedentary behaviour was 79% of monitored hours at age 3 years (IQR 74-84) and 76% (71-80) at age 5 years. Median time spent in moderate to vigorous physical activity represented only 2% of monitored hours at age 3 years (IQR 1-4) and 4% at age 5 years (2-6). Modern British children establish a sedentary lifestyle at an early age. PMID- 14738796 TI - Proportion of tuberculosis transmission that takes place in households in a high incidence area. AB - The prevalence of infection among household contacts of people with tuberculosis is high. This information frequently guides active case finding. We analysed DNA fingerprints of Mycobacterium tuberculosis from 765 tuberculosis patients in Ravensmead and Uitsig, adjacent suburbs of Cape Town, South Africa. In 129 households in which DNA fingerprints were available for more than one patient, we identified 313 patients, of whom 145 (46%) had a fingerprint pattern matching that of another member of the household. The proportion of transmission in the community that took place in the household was 19%, and therefore, in this high incidence area, tuberculosis transmission occurs mainly outside the household. PMID- 14738797 TI - Cholera. AB - Intestinal infection with Vibrio cholerae results in the loss of large volumes of watery stool, leading to severe and rapidly progressing dehydration and shock. Without adequate and appropriate rehydration therapy, severe cholera kills about half of affected individuals. Cholera toxin, a potent stimulator of adenylate cyclase, causes the intestine to secrete watery fluid rich in sodium, bicarbonate, and potassium, in volumes far exceeding the intestinal absorptive capacity. Cholera has spread from the Indian subcontinent where it is endemic to involve nearly the whole world seven times during the past 185 years. V cholerae serogroup O1, biotype El Tor, has moved from Asia to cause pandemic disease in Africa and South America during the past 35 years. A new serogroup, O139, appeared in south Asia in 1992, has become endemic there, and threatens to start the next pandemic. Research on case management of cholera led to the development of rehydration therapy for dehydrating diarrhoea in general, including the proper use of intravenous and oral rehydration solutions. Appropriate case management has reduced deaths from diarrhoeal disease by an estimated 3 million per year compared with 20 years ago. Vaccination was thought to have no role for cholera, but new oral vaccines are showing great promise. PMID- 14738798 TI - Wet markets--a continuing source of severe acute respiratory syndrome and influenza? AB - CONTEXT: Live-animal markets (wet markets) provide a source of vertebrate and invertebrate animals for customers in tropical and subtropical regions of the world. Wet markets sell live poultry, fish, reptiles, and mammals of every kind. Live-poultry markets (mostly chicken, pigeon, quail, ducks, geese, and a wide range of exotic wild-caught and farm-raised fowl) are usually separated from markets selling fish or red-meat animals, but the stalls can be near each other with no physical separation. Despite the widespread availability of affordable refrigeration, many Asian people prefer live animals for fresh produce. Wet markets are widespread in Asian countries and in countries where Asian people have migrated. Live-poultry markets were the source of the H5N1 bird-influenza virus that transmitted to and killed six of 18 people in Hong Kong. STARTING POINT: Yi Guan and colleagues (Science 2003; 302: 276-78) recently reported the isolation of severe acute respiratory syndrome (SARS) coronavirus (CoV) from Himalayan palm civets (Paguna larvata) in wet markets in Shenzen, southern China. These researchers also found serological evidence of infection in raccoon dogs (Nyctereutes procuyoinboides). Serological evidence for SARS CoV in human beings working in these markets, taken together with the earliest cases of SARS in restaurant workers, supports the contention of a potential zoonotic origin for SARS. WHERE NEXT? Will SARS reappear? This question confronts public-health officials worldwide, particularly infectious disease personnel in those regions of the world most affected by the disease and the economic burden of SARS, including China, Taiwan, and Canada. Will the virus re-emerge from wet markets or from laboratories working with SARS CoV, or are asymptomatic infections ongoing in human beings? Similar questions can be asked about a pandemic of influenza that is probably imminent. Knowledge of the ecology of influenza in wet markets can be used as an early-warning system to detect the reappearance of SARS or pandemic influenza. PMID- 14738799 TI - WHO, the Global Fund, and medical malpractice in malaria treatment. PMID- 14738800 TI - Intravenous fluids for seriously ill children. PMID- 14738801 TI - Intravenous fluids for seriously ill children. PMID- 14738803 TI - Intravenous fluids for seriously ill children. PMID- 14738804 TI - HIVNET 012 and Petra. PMID- 14738805 TI - HIVNET 012 and Petra. PMID- 14738806 TI - HIVNET 012 and Petra. PMID- 14738808 TI - HIVNET 012 and Petra. PMID- 14738810 TI - Thought for food in the Philippines. PMID- 14738811 TI - Treatment of multidrug-resistant tuberculosis in Russian prisons. PMID- 14738812 TI - Possible cases of sexual and congenital transmission of sleeping sickness. PMID- 14738813 TI - El Nino and health. PMID- 14738814 TI - The hidden patient. PMID- 14738815 TI - Energy intake and obesity in Japanese patients with type 2 diabetes. PMID- 14738816 TI - Lyme borreliosis. PMID- 14738817 TI - Sonography-guided injection of botulinum toxin in children with cerebral palsy. PMID- 14738818 TI - Underestimation of allergies in elderly patients. PMID- 14738819 TI - 20 most-cited countries in clinical medicine ranked by population size. PMID- 14738821 TI - Tenley Albright. PMID- 14738827 TI - Bankole A Johnson interview. PMID- 14738829 TI - Tooth colour: a review of the literature. AB - OBJECTIVES: To review current knowledge with respect to tooth colour and its measurement. METHODS: 'Medline' database for the period 1966 to the present day and 'ISI Web of Science' database for the period 1974 to the present day were searched electronically with key words tooth, teeth, colour and color. CONCLUSIONS: The colour and appearance of teeth is a complex phenomenon, with many factors such as lighting conditions, translucency, opacity, light scattering, gloss and the human eye and brain influencing the overall perception of tooth colour. The measurement of tooth colour is possible via a number of methods including visual assessment with shade guides, spectrophotometry, colourimetry and computer analysis of digital images. These methods have successfully been used to measure longitudinal tooth colour changes when the dentition has undergone tooth whitening procedures. PMID- 14738830 TI - Clinical evaluation of a novel whitening gel, containing 6% hydrogen peroxide and a standard fluoride toothpaste. AB - OBJECTIVES: The aim of this study was to determine the effect on tooth colour (after 1 and 2 weeks use) of a self-applied tooth-whitening gel containing 6% hydrogen peroxide using a novel applicator, compared to a standard fluoride toothpaste. METHODS: A 2-week, examiner-blind, stratified, parallel design clinical trial was conducted. Efficacy was measured by comparing the Vita shade scores recorded at baseline and after 1 and 2 weeks of product application. To qualify for the study, subjects were required to have at least one upper incisor/canine tooth measuring Vita shade A3 or darker. All upper incisors/canines of qualifying subjects were evaluated for change in Vita shade irrespective of their level of whiteness at baseline, allowing for the whitening gel to be tested on the full range of Vita tooth shades. One hundred and seventeen subjects were divided into two groups, balanced according to their darkest tooth shade. One group followed a 2-week, twice-daily regimen of brushing with toothpaste followed by the application of the hydrogen peroxide whitening gel. The other group brushed twice daily with toothpaste for 2 weeks. RESULTS: Subjects using the whitening gel showed a statistically significant improvement, (p<0.05) in mean tooth shade score of 0.75 and 1.02 units after 1 and 2 weeks, compared to baseline. These improvements from baseline were significantly greater than those observed for the toothpaste only group (0.14 and 0.35 units change after 1 and 2 weeks). CONCLUSIONS: The self-applied tooth-whitening gel containing 6% hydrogen peroxide has been shown to significantly improve the whiteness of teeth after 1 and 2 weeks of product use, compared to the baseline and the toothpaste only group. PMID- 14738831 TI - In vitro evaluation of a novel 6% hydrogen peroxide tooth whitening product. AB - OBJECTIVES: The aims of this study were to evaluate the in vitro tooth whitening effects of a novel 6% hydrogen peroxide containing tooth whitener Xtra White (XW) on extrinsic stain and intrinsic tooth colour and the effects on enamel and dentine microhardness. In addition, to determine the levels of peroxide found in tooth pulp chambers after treatment with XW in vitro. METHODS: Extrinsic stain changes were determined by measuring colour changes of tea stained hydroxyapatite discs after treatment with water, placebo gel, XW or Colgate Simply White (CSW). Intrinsic tooth colour changes were determined by treating extracted human teeth in vitro with a placebo gel, XW or CSW for a simulated two weeks use and measuring colour differences with a chroma meter. Teeth treated with placebo gel and XW were cross-sectioned and the subsurface enamel and dentine was polished and the microhardness determined. The peroxide concentration found in extracted tooth pulp chambers was determined spectrophotometrically. RESULTS: XW gave significantly more extrinsic stain removal (p<0.0001) and an increase in tooth whiteness (p<0.05) than a placebo gel, but was not significantly different to CSW. There were no significant differences in subsurface enamel and dentine microhardness values for the placebo and XW treated teeth. The mean concentration of peroxide found in the tooth pulp chambers was 0.44 mM, which is over 3000 times below the concentration reported to cause pulpal enzyme damage. CONCLUSIONS: XW has been shown to be an effective tooth whitening product in vitro with support for its safety on subsurface enamel and dentine together with its safety on tooth pulp tissue. PMID- 14738832 TI - Evaluation of a 6% hydrogen peroxide tooth whitening gel on enamel and dentine microhardness in vitro. AB - OBJECTIVES: The aims of this study were to evaluate the effects of a novel 6% hydrogen peroxide containing tooth whitener, Xtra White (XW), on enamel and dentine microhardness in vitro. METHODS: Polished human enamel and dentine specimens were prepared and baseline microhardness determined. In study 1, enamel specimens were exposed to 20 min cycles of either water, XW or Sprite Light for up to 28 cycles. In studies 2 and 3, enamel specimens were treated with 20 min cycles of either XW or water and exposed to whole saliva at all other times. In study 3, an additional exposure to a fluoride containing toothpaste was conducted. In total, 28 treatments were conducted in order to simulate a 2 weeks product use. In study 4, dentine specimens were treated as per study 3. Final microhardness measurements were taken and for studies 3 and 4 colour measurements were additionally taken. RESULTS: XW and water gave no statistically significant (p>0.05) changes in enamel and dentine microhardness after 28 treatments. Sprite Light gave a significant (p<0.00002) reduction in enamel microhardness after one 20 min treatment. XW showed significant bleaching of enamel and dentine specimens as compared to the water control. CONCLUSIONS: XW does not have any significant effect on enamel and dentine microhardness. PMID- 14738833 TI - A SEM evaluation of a 6% hydrogen peroxide tooth whitening gel on dental materials in vitro. AB - OBJECTIVES: The aim of this study was to evaluate the effects of a novel 6% hydrogen peroxide gel on the surface morphology of dental gold, amalgam, porcelain and composite. METHODS: Admixed high-copper amalgam and hybrid resin composite specimens were prepared in extracted teeth using standard clinical procedures. Feldspathic porcelain and type III gold specimens were prepared in a mould using standard laboratory procedures. One half of the specimens were covered with nail varnish to serve as the control side, leaving the other half exposed. The specimens were treated with pooled whole saliva (1 h), followed by the peroxide gel (20 min), rinsing with water and returning to saliva. This cycling protocol was continued until a total of 28 treatments with the peroxide gel were completed. The samples were prepared for SEM. RESULTS: There were no observable differences at 200x and 2000x magnifications between the control and peroxide gel treated sides on any of the materials tested. CONCLUSIONS: There were no significant effects of the 6% hydrogen peroxide gel on the surface morphology of any of the dental materials tested. PMID- 14738834 TI - The measurement of degradation and wear of three glass ionomers following peroxide bleaching. AB - OBJECTIVES: To investigate, for three glass ionomer dental materials, the effects of a hydrogen peroxide solution on the elution of key elements, and the effects of a novel 6% hydrogen peroxide tooth whitening gel on subsequent wear rates with a toothpaste slurry. METHODS: Discs specimens of three glass ionomers (ChemFlex, Fuji II and Ketac-Fil) were prepared. One set of discs was exposed to either 6% hydrogen peroxide in phosphate buffer, phosphate buffer, water, a carbonated beverage or 38% orthophosphoric acid for 60 min. The ions eluted from the discs were determined by inductively coupled plasma-optical emmision spectroscopy. A second set of discs were conditioned with either a 6% hydrogen peroxide gel (30 min), water (30 min), a carbonated beverage (2 min) or 38% orthophosphoric acid gel (2 min) followed by a three-body abrasive wear treatment (2 min) with a toothpaste slurry. Changes in height of the samples were measured with a digital comparitor. The conditioning and abrasive cycles were repeated thirteen times. RESULTS: For the elution study, the solutions gave only minor release of sodium except orthophosphoric acid which showed increased elution levels of aluminium, silicon and sodium ions. For the wear study, the orthophosphoric acid gave the highest level of wear whilst for the other treatments there were no statistical significant differences in the level of wear (one-way ANOVA). CONCLUSIONS: A 6% hydrogen peroxide solution did not cause significant dissolution, and a novel 6% hydrogen peroxide tooth whitening gel did not significantly increase the three body wear rate of any of the materials tested. PMID- 14738835 TI - Safety evaluation of a novel whitening gel, containing 6% hydrogen peroxide and a commercially available whitening gel containing 18% carbamide peroxide in an exaggerated use clinical study. AB - OBJECTIVES: The aim of this study was to compare the effect on oral soft tissue of a self-applied tooth whitening gel containing 6% hydrogen peroxide (Xtra White) with a marketed paint-on whitening gel containing 18% carbamide peroxide (Colgate Simply White) after 2-weeks of using products four times daily. METHODS: A 2-week, examiner-blind, stratified, parallel design clinical trial was conducted. Twenty subjects were divided into two groups, balanced according to age and gender. Subjects followed a 2-week, twice-daily regimen of brushing with standard fluoride toothpaste and applied gel product to facial aspects of six upper and six lower incisors/canines, twice in succession with 30 min between applications. Soft tissue examinations were performed on Day 1, before the first application of the test products, and on Day 2, 5, 8 and 15. RESULTS: During the 2-week treatment period, twelve adverse reactions were recorded as potentially attributable to the study products, evenly split between the two test groups. All reports were mild in symptoms and resolved without the need of medical intervention. None of the subjects experiencing an adverse event requested to be withdrawn from the study. CONCLUSIONS: Under the exaggerated use conditions of this test, there was no evidence to suggest that either of the whitening gels produced irritation that was building or developing during the course of the study. It is concluded that both products are safe for their intended use. PMID- 14738836 TI - Plasma-treated polystyrene surfaces: model surfaces for studying cell-biomaterial interactions. AB - Biocompatibility of biomaterials relates, amongst others, to the absence of adverse cellular reactions and modulation of cell adhesion and subsequent responses. With respect to tissue-engineering applications, most materials need to evoke cell adhesion and spreading, while potentially displaying differential cell function. Adhesion has frequently been studied in a controlled fashion, using adhesion-supporting and -inhibiting substrata. The aim of this study is to create a panel of related materials with gradually changing surface characteristics in order to sustain similar individual cell adhesion and spreading, yet different cell population behaviour. A series of polystyrene materials was created with increasing oxygen surface incorporation and, concurrently, decreasing water-contact angles. Individual cells adhered and spread on all surfaces whilst showing well-developed focal adhesions and stress fibres. Cell populations demonstrated a decreased growth on surfaces with lower wettability. The biochemical activity of cell populations was not influenced by the surface treatment, but cell proliferation on surfaces increased with increasing oxygen incorporation. Furthermore, surface coverage with assembled fibronectin matrix was higher on the substrata with higher wettability. Finally, the expression of the adhesion-related proteins cadherin-5, focal adhesion kinase and RhoA was increased on surfaces with higher wettability. Further explorations of the cell biological basis of the observed differential behaviour will give more detailed answers on the rules governing cell-material interactions. PMID- 14738837 TI - Early gene expression analyzed by cDNA microarray and RT-PCR in osteoblasts cultured with water-soluble and low molecular chitooligosaccharide. AB - Chitosan has a variety of biological activities. However, little is known about how chitosan modulates the hard tissue forming cells. When we cultured an osteoblastic cell line in alpha-MEM supplemented with 10% FBS and 0.005% chitooligosaccharide for 3 days, alkaline phosphatase (ALP) activity was significantly high compared with the control culture group (p<0.05). This study was focused on gene expression in osteoblasts cultured with water-soluble chitooligosaccharide. cDNA probes were synthesized from isolated RNA and labeled with fluorescent dye. They were hybridized with Human 1.0((R)) cDNA microarray, and fluorescent signal was analyzed. cDNA microarray analysis revealed that 16 genes were expressed at >/=1.5-fold higher signal ratio levels in the experimental group compared with the control group after 3 days. RT-PCR analysis showed that chitosan oligomer induced an increase in the expression of two genes, CD56 antigen and tissue-type plasminogen activator. Furthermore, the expression of mRNAs for BMP-2 was almost identical in the experimental and control groups after 3 days of culture, but slightly increased after 7 days of culture with chitosan oligomer. These results suggest that a super-low concentration of chitooligosaccharide could modulate the activity of osteoblastic cells through mRNA levels and that the genes concerning cell proliferation and differentiation can be controlled by water-soluble chitosan. PMID- 14738838 TI - Mechanism of apatite formation on wollastonite coatings in simulated body fluids. AB - The formation mechanism of apatite on the surface of wollastonite coating was examined. Plasma-sprayed wollastonite coatings were soaked in a lactic acid solution (pH=2.4) to result in the dissolution of calcium from the coating to form silanol (triple bond Si-OH) on the surface. Some calcium-drained samples were soaked in a trimethanol aminomethane solution (pH=10) for 24h to create a negatively charged surface with the functional group (triple bond Si-O(-)). These samples before and after treatment in a trimethanol aminomethane solution were immersed in simulated body fluids (SBF) to investigate the precipitation of apatite on the coating surface. The results indicate that the increase of calcium in the SBF solution is not the critical factor affecting the precipitation of apatite on the surface of the wollastonite coating and the apatite can only form on a negatively charged surface with the functional group (triple bond Si-O(-)). The mechanism of apatite formation on the wollastonite coating is proposed. After the wollastonite coatings are immersed into the SBF, calcium ions initially exchange with H(+) leading to the formation of silanol (triple bond Si-OH) on the surface of the layer and increase in the pH value at the coating-SBF interface. Consequently, a negatively charged surface with the functional group (triple bond Si-O(-)) forms on the surface. Due to the negatively charged surface, Ca(2+) ions in the SBF solution are attracted to the interface between the coating and solution, thereby increasing the ionic activity of the apatite at the interface to the extent that apatite precipitates on the coating surface. PMID- 14738839 TI - Influence of synthesis and sintering parameters on the characteristics of carbonate apatite. AB - A new method to synthesise carbonate-substituted hydroxyapatite (CHA) powder has been set up introducing a CO(2) flux, as a source of carbonate, in the HA synthesis process based on the neutralisation reaction. The reactants are abundant and inexpensive. The yield is good compared to other CHA powder synthesis. The reaction may be performed at low temperature and without pH control and does not produce any by-products. The influence of the synthesis parameters (temperature, H(3)PO(4) solution dropping rate, i.e. reaction time, CO(2) flux, ageing time) has been tested to optimise the process conditions in order to obtain the highest carbonation degree and favour the B-type CHA precipitation with respect to A-type one. The prepared powder (5.8wt% of total carbonate with an A/B ratio of 0.78) was thermally treated at various temperatures in the range 500-1400 degrees C in different atmospheres (air, wet and dry carbon dioxide). The thermal treatments were performed with a double aim, to eliminate selectively the carbonate groups in A-position maintaining the B type substitution, and to evaluate the thermal stability of the CHA and the total loss of carbonate as a function of temperature. The thermal treatment at 900 degrees C in wet CO(2) gave the best result in terms of a high carbonate residue and a low A/B ratio. We also investigate the use of different techniques (inductively coupled plasma, TGA, Fourier transformed infrared spectroscopy, X ray diffraction) for characterising CHA and calculating sensitivity and accuracy in the quantification of carbonate ions for each molecular site. PMID- 14738840 TI - Preparation and characterization of cationic PLGA nanospheres as DNA carriers. AB - Nanoparticles formulated from biodegradable polymers such as poly(lactic acid) (PLA) and poly(lactide-co-glycolide) (PLGA) are being extensively investigated as non-viral gene delivery systems due to their controlled release characteristics and biocompatibility. PLGA nanoparticles for DNA delivery are mainly formulated by an emulsion-solvent evaporation technique using PVA as a stabilizer generating negatively charged particles and heterogeneous size distribution. The objective of the present study was to formulate cationically modified PLGA nanoparticles with defined size and shape that can efficiently bind DNA. An Emulsion-diffusion evaporation technique to make cationic nanospheres composed of biodegradable and biocompatible co-polyester PLGA has been developed. PVA-chitosan blend was used to stabilize the PLGA nanospheres. The nanospheres were characterized by atomic force microscopy (AFM), photon-correlation spectroscopy (PCS), and Fourier transform infrared spectroscopy (FTIR). Zeta potential and gel electrophoresis studies were also performed to understand the surface properties of nanospheres and their ability to condense negatively charged DNA. The designed nanospheres have a zeta potential of 10mV at pH 7.4 and size under 200nm. From the gel electrophoresis studies we found that the charge on the nanospheres is sufficient to efficiently bind the negatively charged DNA electrostatically. These cationic PLGA nanospheres could serve as potential alternatives of the existing negatively charged nanoparticles. PMID- 14738841 TI - TEM study of calcium phosphate precipitation on bioactive titanium surfaces. AB - Formation of calcium phosphate on alkali- and heat-treated titanium surfaces was investigated using transmission electron microscopy (TEM). The samples were prepared by immersing the alkali- and heat-treated titanium in a revised simulated body fluid (R-SBF) with the same HCO(3)(-) concentration level as in human blood plasma. The deposition and precipitation on treated titanium surfaces were extracted for TEM examination without thinning process. Electron diffraction of the precipitates revealed that octacalcium phosphate (OCP), instead of hydroxyapatite (HA), directly nucleates from amorphous calcium phosphate. The OCP crystals continuously grew on the titanium surfaces rather than transforming into apatite. Calcium titanate was also identified by electron diffraction. Its role in the formation bioactive calcium phosphate, however, is not clear from this experiment. PMID- 14738842 TI - Mutual influence of cholesterol esterase and pseudocholinesterase on the biodegradation of dental composites. AB - It has been demonstrated that human saliva contains cholesterol esterase (CE)- and pseudocholinesterase (PCE)-like hydrolase activities. While PCE has been shown to preferentially degrade triethylene glycol dimethacrylate (TEGDMA) and its derivatives, CE has a greater catalytic effect on the breakdown of bis-phenol A-diglycidyl dimethacrylate (bisGMA) components in composite dental resins. The current study seeks to determine if there is a mutual influence between the different esterases with respect to the biodegradation of resin composite. Photopolymerized model composite resin samples (containing 60% by weight fraction of silanated barium glass filler) based on bisGMA/TEGDMA (bis) or urethane modified bisGMA/TEGDMA/bisEMA (ubis) monomers were incubated in buffer, CE and/or PCE solutions (pH=7.0, 37 degrees C) for 8 and 16 days. The incubation solutions were analyzed for degradation products using high-performance liquid chromatography, UV spectroscopy and mass spectrometry. In the bis system, higher amounts (p<0.05) of a bisGMA derived product, bishydroxy-propoxyphenyl-propane (bisHPPP), were detected in the combined enzyme group as compared to the sum of the two individual enzyme groups. In the ubis system, similar comparisons showed that higher levels (p<0.05) of bisHPPP were detected in the combined group at 8 days while higher amounts (p<0.05) of a bisEMA derived product, ethoxylated bis phenol A, were detected in the combined group at 16 days. The study concluded that CE and PCE act synergistically to increase the biodegradation of both composite resin materials. PMID- 14738843 TI - Long-term stability of bone tissues induced by an osteoinductive biomaterial, recombinant human bone morphogenetic protein-2 and a biodegradable carrier. AB - The long-term stability of bone tissues induced by recombinant human bone morphogenetic protein-2 (rhBMP-2) and poly[L-lactide-co-glycolide] copolymer coated gelatin sponge (PGS) was examined. In 16 dogs, 2.5 cm unilateral bone defects were created in the left tibial diaphyses. Tibia was fixed with metal plate, and PGS impregnated with (0.4 mg/cm(3)) or without rhBMP-2 was implanted into 15 or one defects, respectively. The metal plates of rhBMP-2-treated limbs were removed 16 weeks after the implantation. The bilateral tibiae of five animals each of the rhBMP-2-treated group were harvested at 32, 52 or 104 weeks, and served for biomechanical testing and histology. Although the defect that received PGS alone resulted in nonunion at 16 weeks, all defects treated with rhBMP-2 achieved radiographic bony union by 8 weeks. Biomechanical properties of the regenerated bones restored to the levels of intact tibiae at 32 weeks, but torsional stiffness was significantly higher. No statistical significances were detected in all parameters between regenerated and intact tibiae at 104 weeks. No radiographic and histological findings suggesting enhanced resorption to the regenerated bones were observed. These results suggest the long-term stability of the bone tissues induced by rhBMP-2, and the usefulness of rhBMP-2-impregnated PGS as a biomaterial for long bone defect filling. PMID- 14738844 TI - Glow discharge plasma pretreatment enhances osteoclast differentiation and survival on titanium plates. AB - Despite the fact that several reports have demonstrated osteoclast activity on various bioactive ceramics, osteoclast functions on surface-modified titanium have not come under focus. This study aimed to examine whether the increasing surface energy of glow discharge plasma (GDP) involved in protein adhesion containing the RGD (Arg-Gly-Asp) sequence affects osteoclast responses on titanium plates. We examined osteoclast differentiation and survival rates on titanium plates with and without GDP. The amounts of osteoclasts on titanium plates were not increased by GDP after 1 week. However, osteoclast differentiation was greatly activated by GDP pretreatment, as tartrate-resistant acid phosphatase synthesis significantly increased on the titanium plates with GDP. Additionally, since the presence of osteoclasts was detected only on the titanium plates with GDP, even after 4h cultivation in a coculture test, the osteoclasts survival rate was increased by GDP pretreatment. As osteoclast responses were affected even on surface modified metallic materials, we concluded that novel approaches are needed not only for osteoclastic resorption on ceramic materials but also for osteoclast responses on surface-modified metallic materials. PMID- 14738845 TI - Hepatocyte behavior on synthetic glycopolymer matrix: inhibitory effect of receptor-ligand binding on hepatocyte spreading. AB - The interaction of carbohydrate-based polymers with asialoglycoprotein receptors (ASGPRs) on the surface of hepatocytes has been used to design hepatocyte adhesion matrices. Therefore, we have characterized the interaction of ASGPR on the surface of hepatocytes with glycopolymer-coated surfaces. Since ASGPRs bound to glycopolymer surfaces escape from internalization and degradation, they were quantified by western blot analysis. The amount of hepatocyte ASGPRs that initially adhered to the glycopolymer surface was proportional to the concentration of the coated glycopolymer. We found that the initial adhesion of hepatocytes to the glycopolymer surface was enhanced by interactions with ASGPR, whereas interactions with ASGPR inhibited the post-adhesion process, a cell adhesion phenomenon that occurs following the initial adhesion. Furthermore, hepatocytes are much more spread on glycopolymer surfaces with lower coating density. Taken together, we suggest that the post-adhesion process triggered hepatocyte spreading on glycopolymer surfaces, and ASGPR-carbohydrate interactions act negatively on the post-adhesion mechanism as well as on hepatocyte spreading on glycopolymer surfaces depending on the density of coated glycopolymers. PMID- 14738846 TI - An amino acid-modified and non-HEMA containing glass-ionomer cement. AB - It is known that unreacted 2-hydroxyethyl methacrylate (HEMA) in current resin modified glass ionomer cements (RMGICs) shows potential cytotoxicity to pulp and surrounding tissues. Elimination of HEMA could make RMGICs more attractive for dental applications. In this research, novel six acrylate and methacrylate derivatives of amino acids were synthesized, characterized and used for replace HEMA in RMGICs. The experimental RMGICs were formulated with vinyl-containing polymer, amino acid derivative, water, and commercial Fuji II LC glass. Among all the derivatives, methacryloyl beta-alanine (MBA) was selected for further formulations due to its relatively low solution viscosity and high CS. Effects of polymer content and powder/liquid, P/L, ratio were significant. The formulation with liquid composition of 50/25/25 (polymer/MBA/water) and P/L ratio of 2.7/1 was found the optimal. It appears that this novel non-HEMA-containing RMGIC system based on amino acid derivatives will be a better dental restorative because it demonstrated improved mechanical strengths and may eliminate potential cytotoxicity in current RMGICs caused by leached HEMA. The optimal MBA-modified GIC were 20% higher in CS, 70% higher in DTS and 93% higher in FS, compared to Fuji II LC. PMID- 14738847 TI - Healing of periodontal defects treated with enamel matrix proteins and root surface conditioning--an experimental study in dogs. AB - Application of enamel matrix proteins has been introduced as an alternative method for periodontal regenerative therapy. It is claimed that this approach provides periodontal regeneration by a biological approach, i.e. creating a matrix on the root surfaces that promotes cementum, periodontal ligament (PDL) and alveolar bone regeneration, thus mimicking the events occurring during tooth development. Although there have been numerous in vitro and in vivo studies demonstrating periodontal regeneration, acellular cementum formation and clinical outcomes via enamel matrix proteins usage, their effects on the healing pattern of soft and hard periodontal tissues are not well-established and compared with root conditioning alone. In the present study, the effects of Emdogain (Biora, Malmo, Sweden), an enamel matrix derivative mainly composed of enamel matrix proteins (test), on periodontal wound healing were evaluated and compared with root surface conditioning (performed with 36% orthophosphoric acid) alone (control) histopathologically and histomorphometrically by means of the soft and hard tissue profile of periodontium. An experimental periodontitis model performed at premolar teeth of four dogs were used in the study and the healing pattern of periodontal tissues was evaluated at days 7, 14, 21, 28 (one dog at each day), respectively. At day 7, soft tissue attachment evaluated by means of connective tissue and/or epithelial attachment to the root surfaces revealed higher connective tissue attachment rate in the test group and the amount of new connective tissue proliferation in the test group was significantly greater than the control group (p<0.01). New bone formation by osteoconduction initiated at day 14 in the test and control group. At day 21, the orientation of supra alveolar and PDL fibers established, and new cementum formation observed in both groups. At day 28, although regenerated cementum was cellular in all of the roots in the control samples, an acellular type of cementum (1.32+/-0.83 mm in length and 3.16+/-0.23 microm in width) was also noted in six roots of test samples with an inconsistent distribution on the root surfaces. The amount of new cementum was significantly higher in the test group than the control group samples (p<0.01). The width of the cellular cementum in the control group was more than the cellular cementum in the test group, but the difference was not statistically significant (p>0.05). A firm attachment of acellular cementum to the root dentin with functional organization of its collagen fibers was noted, and, the accumulation and organization of cellular cementum in the control group was more irregular than the cellular cementum formed in the test group. The amount of new bone was 2.41+/-0.75 mm in the test and 1.09+/-0.46 mm in the control group at day 28. The rate of bone maturation (the number of osteons) was found higher in the test group (10.75+/-0.85) than the control group (5.50+/-0.86). Under the limitations of the study, our results reveal that when compared with root surface conditioning, enamel matrix proteins have more capacity for stimulating periodontal regeneration via their positive effects on root surfaces, i.e. inhibition of gingival epithelium down growth and stimulation of connective tissue proliferation and attachment to the root surfaces during wound healing. An acellular type of cementum regeneration and new alveolar bone formation by an accelerated osteoconductive mechanism are also achieved with application of enamel matrix proteins. PMID- 14738848 TI - Engineering a bifunctional starch-cellulose cross-bridge protein. AB - Biodegradable starch- and cellulose-based polymers have a range of properties which make them suitable for use in a wide array of biomedical applications ranging from bone replacement to engineering of tissue scaffolds and drug delivery systems. A novel polysaccharide cross-bridging protein was designed which was comprised of a cellulose-binding domain from Clostridium cellulovorans (CBD(clos)) and a starch-binding domain from Aspergillus niger B1 (SBD(Asp)). The two genes were fused in-frame via a synthetic elastin gene to construct a Cellulose/Starch Cross bridging Protein (CSCP). Recombinant CSCP was expressed in Escherichia coli, and successfully refolded from inclusion bodies. CSCP demonstrated cross-bridging ability in different model systems composed of insoluble or soluble starch and cellulose. The aspect that different carbohydrate binding module maintain their binding capacity over a wide range of conditions, without the need for chemical reactions, makes them attractive domains for designing new classes of chimeric polysaccharide-binding domains which demonstrate potential for use in a wide range of biomaterials. PMID- 14738849 TI - Microcontact printing of proteins on oxygen plasma-activated poly(methyl methacrylate). AB - This paper describes a method for microcontact printing protein solutions onto polymer substrates temporarily activated by oxygen plasma. Following plasma treatment, poly(dimethyl siloxane) (PDMS) stamps were coated with an aqueous laminin solution then placed in direct contact with plasma-treated poly(methyl methacrylate) (PMMA) substrates. This process resulted in well defined laminin stripes on the PMMA surface when printing was performed within 45min of the plasma treatment. Axonal outgrowth from embryonic chick dorsal root ganglia (DRG) was largely confined to the stamped pattern, while over 90% of primary rat Schwann cells adhered to the protein stamped areas on the PMMA substrates. Oxygen plasma treatment of the PMMA surface was necessary to deposit proteins that direct axonal outgrowth from chick DRG and Schwann cell adherence. PMID- 14738850 TI - Surface tailoring of poly(DL-lactic acid) by ligand-tethered amphiphilic polymer for promoting chondrocyte attachment and growth. AB - The ligand-tethered poly(ethylene oxide-propylene oxide-ethylene oxide) (PEO-PPO PEO) triblock copolymer was explored to engineer poly(DL-lactic acid) (PDL-LA) material to promote cell attachment and growth. The PEO-PPO-PEO was activated by methyl sulfonyl chloride and the amino acid, and peptide were attached. By blending the PDL-LA with the ligand-tethered PEO-PPO-PEO derivatives, the surface of modified PDL-LA film was investigated by ATR-FTIR, XPS and contact angle. The chondrocytes test on different PDL-LA films indicated that the PEO-PPO-PEO amino acid and RGD derivatives modified PDL-LA films could promote chondrocyte attachment and growth. This simple surface treatment method may have potentials for tissue engineering and other biomedical applications. PMID- 14738851 TI - Neointima formation on vascular elastic laminae and collagen matrices scaffolds implanted in the rat aortae. AB - Synthetic polymers, including polytetrafluoroethylene and Dacron, and biomatrix proteins, including collagen and fibrin, have been used for the construction of vascular substitutes. However, these materials induce inflammatory reactions, contributing to thrombosis, smooth muscle cell (SMC) proliferation, and neointima formation, processes leading to the failure of vascular substitutes. Thus, a pressing issue in vascular reconstruction is to construct vascular substitutes with surface materials that are inflammation-resistant. Here, we demonstrate that the vascular elastic laminae exhibit such a property. Aortic specimens from donor rats were treated with 0.1M NaOH for various times, resulting in elastic lamina collagen matrix scaffolds with and without the basal lamina. Matrix scaffolds were implanted into the host aorta with three different surface materials, including the elastic lamina, basal lamina, and adventitial collagen, and observed for leukocyte adhesion, endothelial migration, cell proliferation, and neointimal formation on these surfaces. It was found that the elastic lamina was associated with significantly lower leukocyte adhesion, BrdU incorporation, and neointima formation than the basal lamina and adventitial collagen, while the migration of endothelial cells was comparable on all three surfaces. The adventitial collagen matrix was associated with leukocyte infiltration from blood and subsequent SMC migration from the host aorta, whereas the elastic laminae were resistant to such processes. The morphology of the implanted elastic laminae appeared normal at all times. These observations suggest that the vascular elastic laminae exhibit inflammation-resistant properties and inhibit SMC mitogenic activities compared with collagen-containing matrices and may be considered a potential surface material for vascular reconstruction. PMID- 14738852 TI - Electrospun P(LLA-CL) nanofiber: a biomimetic extracellular matrix for smooth muscle cell and endothelial cell proliferation. AB - Poly(L-lactide-co-epsilon-caprolactone) [P(LLA-CL)] with L-lactide to epsilon caprolactone ratio of 75 to 25 has been electrospun into nanofibers. The relationship between electrospinning parameters and fiber diameter has been investigated. The fiber diameter decreased with decreasing polymer concentration and with increasing electrospinning voltage. The X-ray diffractometer and differential scanning colorimeter results suggested that the electrospun nanofibers developed highly oriented structure in CL-unit sequences during the electrospinning process. The biocompatibility of the nanofiber scaffold has been investigated by culturing cells on the nanofiber scaffold. Both smooth muscle cell and endothelial cell adhered and proliferated well on the P(LLA-CL) nanofiber scaffolds. PMID- 14738853 TI - Fabrication of nano-structured porous PLLA scaffold intended for nerve tissue engineering. AB - Nerve tissue engineering (NTE) is one of the most promising methods to restore central nerve systems in human health care. Three-dimensional distribution and growth of cells within the porous scaffold are of clinical significance for NTE. In this study, an attempt was made to develop porous polymeric nano-fibrous scaffold using a biodegradable poly(L-lactic acid) (PLLA) for in vitro culture of nerve stem cells (NSCs). The processing of PLLA scaffold has been carried out by liquid-liquid phase separation method. The physico-chemical properties of the scaffold were fully characterized by using differential scanning calorimetry and scanning electron microscopy. These results confirmed that the prepared scaffold is highly porous and fibrous with diameters down to nanometer scale. As our nano structured PLLA scaffold mimics natural extracellular matrix, we have intended this biodegradable scaffold as cell carrier in NTE. The in vitro performance of NSCs seeded on nano-fibrous scaffold is addressed in this study. The cell cultural tests showed that the NSCs could differentiate on the nano-structured scaffold and the scaffold acted as a positive cue to support neurite outgrowth. These results suggested that the nano-structured porous PLLA scaffold is a potential cell carrier in NTE. PMID- 14738854 TI - Importance of integrin beta1-mediated cell adhesion on biodegradable polymers under serum depletion in mesenchymal stem cells and chondrocytes. AB - To evaluate the predominant mechanism of chondrogenic cell [mesenchymal stem cells (MSCs) and chondrocytes] adhesion under serum free conditions, we measured the surface roughness and wettability of poly(lactic acid:polyglycolic acid=75:25) (PLGA), poly(lactic acid) (PLA), and poly(-epsilon-caprolactone) (PCL)-coated glass plates. Also to evaluate the biological reactions involved in cell-polymer interactions, integrin beta1, one of the cell adhesion molecules, was blocked with monoclonal antibody. In cell attachment test, MSCs and chondrocytes adhesion to synthetic polymers in 1h were very low and ranged from 2.8% to 8.0%. In present study, the correlation between attachment rate and surface roughness, contact angle, or integrin beta1 blocking on PLGA, PLA and PCL coated plates could not be proved. However, we found that L-arginine-coated PLA highly increased the attachment rates of MSCs (30.2%) and of chondrocytes (26%), whereas integrin beta1 blocking significantly decreased these attachment rates to 5.6% and 7.4%, respectively, suggesting that increased cell adhesion to L arginine-coated plates is mediated by integrin beta1. In this study, we showed that polymer characteristics such as roughness and wettability did not play an important role in cell adhesion under serum free conditions, because there was no significant difference according to polymer characteristics, whereas biological interactions mediated by integrin beta1 were critical during the early period of cell adhesion. The results suggest that L-arginine could be useful for facilitating early cell adhesion to synthetic polymers in cartilage tissue engineering. PMID- 14738855 TI - Dermal wound healing processes with curcumin incorporated collagen films. AB - The wound healing process involves extensive oxidative stress to the system, which generally inhibits tissue remodeling. In the present study, an improvement in the quality of wound healing was attempted by slow delivery of antioxidants like curcumin from collagen, which also acts as a supportive matrix for the regenerative tissue. Curcumin incorporated collagen matrix (CICM) treated groups were compared with control and collagen treated rats. Biochemical parameters and histological analysis revealed that increased wound reduction, enhanced cell proliferation and efficient free radical scavenging in CICM group. The higher shrinkage temperature of CICM films suggests increased hydrothermal stability when compared to normal collagen films. Spectroscopic studies revealed that curcumin was bound to the collagen without affecting its triple helicity. Further we adopted the antioxidant assay using 2,2'-azobisisobutyronitrile to assess in vitro antioxidant activity of CICM. The antioxidant studies indicated that CICM quenches free radicals more efficiently. This study provides a rationale for the topical application of CICM as a feasible and productive approach to support dermal wound healing. PMID- 14738856 TI - Mechanistic evaluation of the glucose-induced reduction in initial burst release of octreotide acetate from poly(D,L-lactide-co-glycolide) microspheres. AB - One major obstacle for development of injectable biodegradable microspheres for controlled peptide and protein delivery is the high initial burst of drug release occurring over the first day of incubation. We describe here the significant reduction in initial burst release of a highly water-soluble model peptide, octreotide acetate, from poly(D,L-lactide-co-glycolide) microspheres by the co encapsulation of a small amount of glucose (e.g., 0.2%w/w), i.e., from 30+/-20% burst - glucose to 8+/-3% + glucose (mean+/-SD, n=4). This reduction is unexpected since hydrophilic additives are known to increase porosity of microspheres, causing an increase in permeability to mass transport and a higher burst. Using the double emulsion-solvent evaporation method of encapsulation, the effect of glucose on initial burst in an acetate buffer pH 4 was found to depend on polymer concentration, discontinuous phase/continuous phase ratio, and glucose content. Extensive characterization studies were performed on two microsphere batches, +/-0.2% glucose, to elucidate the mechanism of this effect. However, no significant difference was observed with respect to specific surface area, porosity, internal and external morphology and drug distribution. Continuous monitoring of the first 24-h release of octreotide acetate from these two batches disclosed that even though their starting release rates were close, the microspheres + glucose exhibited a much lower release rate between 0.2 and 24h compared to those - glucose. The microspheres + glucose showed a denser periphery and a reduced water uptake at the end of 24-h release, indicating decreased permeability. However, this effect at times was offset as glucose content was further increased to 1%, causing an increase in surface area and porosity. In summary, we conclude that the effect of glucose on initial burst are determined by two factors: (1) increased initial burst due to increased osmotic pressure during encapsulation and drug release, and (2) decreased initial burst due to decreased permeability of microspheres. PMID- 14738857 TI - A one-pot synthesis of oleic acid end-capped temperature- and pH-sensitive amphiphilic polymers. AB - A one-pot synthesis of an amphiphilic oleic acid-end capped random poly(N isopropylacrylamide-co-N,N-dimethylacrylamide) (2) is reported. In aqueous media, the solubility of 2 was temperature- and pH-sensitive. Both the lower critical solution temperature and critical micelle concentration (CMC) of 2 were pH dependent. The LCST of 2 was around 35.2 degrees C in acidic buffer solutions (pH=2.00-5.00), and it was increased significantly to around 38.4 degrees C in neutral and alkaline buffer solutions (pH 7.00-11.00). Polymer 2 exhibited a phase transition pH around 6.7, below which the polymer became significantly less water-soluble. The CMC of 2 was 40 mg/l in pH 2.0 buffer solution, and it was increased markedly to 60-67 mg/l in pH 7.0 and pH 11.0 buffer solutions. Micelle solutions of 2 in different pH conditions were prepared by a membrane-dialysis method. In aqueous solution, dynamic light scattering studies revealed that the size of micelles was 50-90 nm with the particle size being larger in acidic solutions. In solid state, transmission electron microscope studies showed that micelles were roughly spherical, their sizes were 25-90 nm and it decreased with the increase of solution pH. The pH-sensitivity of 2 was triggered by the -COOH in the hydrophobic segment. The temperature- and pH-sensitivity of the novel polymeric micelles would make an interesting drug delivery system. PMID- 14738858 TI - Enzyme immobilization in novel alginate-chitosan core-shell microcapsules. AB - Alginate-chitosan core-shell microcapsules were prepared in order to develop a biocompatible matrix for enzyme immobilization, where the protein is retained either in a liquid or solid core and the shell allows permeability control over substrates and products. The permeability coefficients of different molecular weight compounds (vitamin B2, vitamin B12, and myoglobin) were determined through sodium tripolyphosphate (Na-TPP)-crosslinked chitosan membrane. The microcapsule core was formed by crosslinking sodium alginate with either calcium or barium ions. The crosslinked alginate core was uniformly coated with a chitosan layer and crosslinked with Na-TPP. In the case of calcium alginate, the phosphate ions of Na-TPP were able to extract the calcium ions from alginate and liquefy the core. A model enzyme, beta-galactosidase, was immobilized in the alginate core and the catalytic activity was measured with o-nitrophenyl-beta-D galactopyranoside (ONPG). Change in the activity of free and immobilized enzyme was determined at three different temperatures. Na-TPP crosslinked chitosan membranes were found to be permeable to solutes of up to 17,000Da molecular weight. The enzyme loading efficiency was higher in the barium alginate core (100%) as compared to the calcium alginate core (60%). The rate of ONPG conversion to o-nitrophenol was faster in the case of calcium alginate-chitosan microcapsules as compared to barium alginate-chitosan microcapsules. Barium alginate-chitosan microcapsules, however, did improve the stability of the enzyme at 37 degrees C relative to calcium alginate-chitosan microcapsules or free enzyme. This study illustrates a new method of enzyme immobilization for biotechnology applications using liquid or solid core and shell microcapsule technology. PMID- 14738859 TI - Hemocompatibility of polyacrylonitrile dialysis membrane immobilized with chitosan and heparin conjugate. AB - Chitosan (CS)/heparin (HEP) polyelectrolyte complex (PEC) was covalently immobilized onto the surface of polyacrylonitrile (PAN) membrane. The effect of surface modification on the protein adsorption and platelet adhesion, metabolites permeation and anticoagulation activity of the resulting membrane was investigated. Surface characterization such as water contact angle, and X-ray photoelectron spectroscope were performed. The immobilization of PEC caused the water contact angle to reduce, thereby indicating the increase in the hydrophilicity. Protein adsorption, platelet adhesion, and thrombus formation were all reduced by the immobilization of HEP. Anticoagulant activity was evaluated with activated partial thrombin time (APTT), prothrombin time (PT), fibrinogen time, and thrombin time (TT). The results revealed that PEC immobilizing membrane can improve antithrombogenicity of PAN membrane. In addition, the PEC-immobilized membranes can suppress the proliferation of Pseudomonas aeruginosa. In vitro cytotoxicity test showed leachable substance released was below cytotoxic level. The pure water permeability results show little variation due to PEC-immobilization. Thus PEC-immobilization can endow the PAN membrane hemocompatibility and antibacterial activity while retaining the original permeability. PMID- 14738860 TI - Ultrastructural characterization of the implant/bone interface of immediately loaded dental implants. AB - Primary stability and an optimized load transfer are assumed to account for an undisturbed osseointegration process of implants. Immediate loaded newly designed titanium dental implants inserted in the mandible of minipigs were used for the characterization of the interfacial area between the implant surface and the surrounding bone tissue during the early healing phase. Histological and electron microscopical studies were performed from implant containing bone specimens. Two different load regimens were applied to investigate the load related tissue reaction. Histological and electron microscopical analysis revealed a direct bone apposition on the implant surfaces, as well as the attachment of cells and matrix proteins in the early loading phase. A striking finding of the ultrastructural immunocytochemical investigations was the synthesis and deposition of bone related proteins (osteonectin, fibronectin, fibronectin receptor) by osteoblasts from day one of bone/biomaterial interaction. Calcium-phosphate needle-like crystallites were newly synthesized in a time-related manner directly at the titanium surface. No difference in the ultrastructural appearance of the interface was found between the two loading groups. Our experimental data suggest that loading of specially designed implants can be performed immediately after insertion without disturbing the biological osseointegration process. PMID- 14738861 TI - Biomimetic glucose recognition using molecularly imprinted polymer hydrogels. AB - Non-covalent molecular imprinting of poly(allylamine hydrochloride) (PAA.HCl) with D-glucose 6-phosphate monobarium salt (GPS-Ba) produced molecularly imprinted polymer hydrogels (MIP) having an affinity to glucose over fructose. The hydrogels were formed by ionic association of the template molecule, GPS-Ba, to the polymer, prior to covalent crosslinking using epichlorohydrin (EPI). The template was removed by an aqueous base wash. Batch equilibration studies using different MIP hydrogels and non-molecularly imprinted polymers (NIPs) were performed in aqueous and buffered media to determine the binding capacities and isomeric selectivities with respect to the sugars, glucose and fructose. MIP glucose hydrogels exhibited binding capacities in excess of 0.6g of glucose per g of dry gel in a 100% DI H(2)O glucose solution, and in a 50-50% glucose-fructose solution mixture. Equilibrium binding capacities of fructose were lower than those observed with respect to glucose, indicating an isomeric preference for the binding of glucose over fructose. These hydrogels demonstrated a remarkable degree of biomimetic sugar recognition to specifically and selectively bind glucose in their swollen state in environments mimicking physiological conditions. PMID- 14738862 TI - Contrasting roles for CXCR2 during experimental colitis. AB - Neutrophil recruitment into the colon is believed to play a crucial pathogenic role in the progression of clinical and experimental inflammatory bowel diseases (IBDs). The chemokine receptor CXCR2 is highly expressed on neutrophils, and promotes neutrophil recruitment in several inflammatory diseases. The present study determined the biological role of CXCR2 during trinitrobenzene sulfonic acid (TNBS)-induced colitis in the rat by assessing effects of CXCR2 antibody neutralization on neutrophil accumulation during the early (8 h) and late phase (day 7) of TNBS-induced colitis. CXCR2 expression was elevated (>3-fold above control) within 8 h and remained elevated to day 7 of colitis induction, in parallel with significant increases in neutrophil infiltration. Treatment of colitic rats with a single dose of CXCR2 neutralizing antibody significantly reduced colonic neutrophil accumulation during the early (8 h) phase of TNBS induced colitis. However, chronic administration of CXCR2 antibody did not reduce colonic neutrophil accumulation during the late phase (day 7) of TNBS-induced colitis. In summary, the present findings suggest a functional role for CXCR2 in initiating neutrophil recruitment during the early phase of TNBS-induced acute colitis, and demonstrate that: early colonic neutrophil accumulation is CXCR2 dependent and the late phase colonic neutrophil accumulation is CXCR2 independent. PMID- 14738863 TI - The proteasome inhibitor, PS-341, causes cytokeratin aggresome formation. AB - Mallory body (MB) experimental induction takes 10 weeks of drug ingestion. Therefore, it is difficult to study the dynamics and mechanisms involved in vivo. Consequently, an in vitro study was done using primary tissue culture of hepatocytes from drug-primed mice livers in which MBs had already formed. The hypothesis to be tested was that MBs are cytokeratin aggresomes, which form when hepatocytes have a defective ubiquitin-proteasome pathway by which turnover of cytokeratin proteins is prevented. To test this hypothesis, primary tissue cultures of the hepatocytes from normal and MB-forming livers were incubated with the proteasome inhibitor PS-341 and then the cytokeratin filaments and the filament connecting proteins, that is, beta-actin, and ZO1, were visualized by immunofluorescence microscopy. PS-341 caused detachment of the cytokeratins from the cell surface plasma membrane. The cytokeratin filaments retracted toward the nucleus and cytokeratin aggresomes formed. In human livers, MBs showed colocalization of cytokeratin-8 (CK-8) with ubiquitin but not with beta-actin or ZO1. Mouse hepatoma cell lines were studied using PS-341 to induce cytokeratin aggresome formation. In these cell lines, the cytokeratin filaments first retracted toward the nucleus then formed cytokeratin-ubiquitin aggresomes polarized at one side of the nucleus. At the same time, the cells became dissociated from each other, however. The results simulated MB formation. MBs differ from cytokeratin aggresomes both morphologically and in ultrastructure. PMID- 14738864 TI - C5b-9 terminal complement complex assembly on apoptotic cells in human arterial wall with atherosclerosis. AB - Apoptosis plays an important role in atherosclerosis. The factors regulating this process are not well defined. We examined the relation of apoptotic cells with the terminal complement complex C5b-9 in human atherosclerotic lesions. The extent of apoptosis was determined using TdT dUTP nick-end labeling (TUNEL) and immunohistochemistry of apoptosis regulators caspase-3, caspase-9, Bax, and Bcl 2. C5b-9 was localized by immunohistochemistry and immunoelectron microscopy. The apoptotic index was higher in fibrous plaques when compared with intimal fatty streaks and intimal thickenings. Bax expression was present in TUNEL+ apoptotic cells, and Bcl-2 was rarely present in the atherosclerotic wall. Active caspase 9 and caspase 3 deposits were present in the same areas, suggesting an involvement of the mitochondrial pathway. C5b-9 deposits colocalized with TUNEL+ cells, and the percent of double-positive cells was 2% in fatty streaks, 12% in intimal thickenings, and 35% in fibrous plaques. Colocalization of apoptotic cells with C5b-9 was also confirmed by immunoelectron microscopy. In conclusion, some apoptotic cells carry C5b-9 deposits, suggesting that complement might be activated by apoptotic cells and involved in the promotion of apoptosis, contributing to the progression of atherosclerotic lesions. PMID- 14738865 TI - RXRalpha mRNA expression is associated with cell proliferation and cell cycle regulation in Hep3B cell. AB - Retinoids are well-characterized differentiation and anti-proliferation agents. The functional role of retinoid x receptor alpha (RXRalpha) in cell proliferation and cell cycle regulation is not well understood. Using human Hep3B cell line, we showed that the mRNA level of RXRalpha was closely associated with cell growth. RXRalpha mRNA expression elevated along with the proliferation of Hep3B cells. This association was most evident in RXRalpha and was also noted with retinoic acid (RA) receptor alpha (RARalpha), but not found in other RARs and RXRs. The expression of RXRalpha and cyclin A mRNA was co-regulated when Hep3B cells were cultured in serum-free medium. The mRNA levels of RXRalpha and cyclin A appeared to be highest in G1/S phase in Hep3B cells treated by aphidicolin. Taken together, our data suggest that RXRalpha may be actively involved in cell proliferation and cell cycle regulation in Hep3B cells. PMID- 14738866 TI - Description of a flow cytometry approach based on SYBR-14 staining for the assessment of DNA content, cell cycle analysis, and sorting of living normal and neoplastic cells. AB - RATIONALE: This study aimed to expand the utilization of a simplified flow cytometric approach that employing SYBR-14/PI staining into broader flow cytometry applications, including (i) measurement of the DNA content; (ii) performing cell cycle analysis on mammalian cells; and (iii) sorting of live SYBR 14-stained mammalian cells based on DNA content. MATERIAL AND METHODS: Cell lines of human origin were stained with SYBR-14 and propidium iodide (PI) and assessed by a dual-color flow cytometry. Finally, sorting of living SYBR-14-stained human cell lines was performed. RESULTS: Dual staining with SYBR-14 and PI of human cells followed by flow cytometry analysis demonstrates that in addition to quality assessment, this staining could be utilized to determinate the DNA content on mammal cells. In addition, it resolves the diploid, tetraploid, and aneuploid DNA content. Furthermore, the SYBR-14-stained mammal cells were efficiently sorted based on DNA content and live cells were obtained. All these features have not been previously described with the utilization of this staining approach. CONCLUSIONS: Results of this study demonstrate that this flow cytometric approach not only allows assessment of the viability of cells, but also the DNA content of mammal cells. In addition, this approach allows one to sort viable cells stained with SYBR-14. These findings open-up unexpected and unrestricted avenues for sorting of living mammal cells and provide significant advantages over the traditionally cumbersome sorting approaches for living cells, which demand very specialized and expensive UV light sources as well as sophisticated sorting procedures. PMID- 14738867 TI - Gene expression in human osteoblastic cells from normal and heterotopic ossification. AB - Heterotopic ossification (HO), a possible complication of head injury, develops in sites where it is not normally present like at the vicinity of joints. It may cause pain, decrease motion and in severe cases complete joint ankylosis requiring surgical intervention. To our knowledge, no study has been made to analyze HO at the molecular level on human biopsies, whereas its etiology remains to be determined. We defined a procedure of cell fractionation from bone resections and developed quantitative RT-PCR to compare genetic expression patterns between human normal osteoblasts and heterotopic ossification forming cells. This quantitative study demonstrated a specific and strong overexpression of osteocalcin mRNA in HO-isolated cells associated with a significant upregulation of type 1 collagen and osteonectin mRNA while histological analysis showed only small cellular variations. Our results give a first molecular characterization of heterotopic ossification and we conclude that such overexpressions in HO-isolated cells could be associated with the high activity of this pathological bone. PMID- 14738868 TI - Transgenic mice expressing hepatitis B virus X protein are more susceptible to carcinogen induced hepatocarcinogenesis. AB - The hepatitis B virus X (HBx) protein is thought to be implicated in the development of human hepatocellular carcinoma (HCC), but its exact function remains controversial. To investigate whether the expression of the HBx gene alone can induce HCC on an inbred C57BL/6 strain that displays a lower spontaneous rate of liver cancer, and to determine if HBx transgenic mice are more susceptible to the effects of hepatocarcinogens, C57-TgN (HBx) X transgenic mice were bred with normal C57BL/6 mice strain. The F1 mice (about 50% HBx positive and 50% HBx negative) were treated with a single dose of diethylnitrosamine (DEN) at 7 days of age, or were untreated. Mice were killed at appropriate time points and were analyzed for histological change in the liver. The expression of HBx protein were examined by using immunohistochemical staining. Glycogen storage foci were examined by using periodic acid-Schiff (PAS) staining. In HBx transgenic mice untreated with DEN, HBx expression and glycogen storage foci were always observed in the liver after 8 weeks, but not obvious histological pathologic changes. Histological examination of liver tissue confirmed that DEN-treated HBx mice developed approximately twice as many focal lesions of basophilic hepatocytes as treated wild-type littermates. Hepatocellular adenomas and carcinomas were also more frequent in DEN-treated HBx positive than HBx-negative mice. Taken together, our results suggest that HBx gene expression alone is not sufficient for carcinogenesis, but may act as a promoter for malignant transformation. PMID- 14738869 TI - Islet cell tumor arising from a heterotopic pancreas in the duodenal wall with ulceration. AB - A rare case of symptomatic islet cell tumor arising from heterotopic pancreas in the duodenum with ulceration is described. Gastrointestinal bleeding was the only sign observed in this patient. Tagged red blood cell scan, upper endoscopy, and computed tomography scan showed active bleeding ulcer from a periampullary mass. Removal of the submucosal tumor was done to prevent future re-bleeding. Histologic and immunohistochemical characterization of the tumor showed an endocrine tumor that expressed a variety of endocrine peptides. PMID- 14738870 TI - Contradictory effects of short- and long-term hyperglycemias on ischemic injury of myocardium via intracellular signaling pathway. AB - Although clinical diabetes mellitus is obviously a high risk factor for myocardial infarction, there is disagreement about the sensitivity of ischemic injury of an infarcted myocardium in experimental studies. The present study evaluated the influences of different durations of hyperglycemia on ischemic and reperfusion injuries of the myocardium, and focused on extracellular signal regulated kinase 1/2 (ERK1/2), which plays an important role in the intracellular signaling pathway and is reported to be associated with myocardial protection against heart injury. Short- and long-term hyperglycemias were induced in rats by streptozotocin (STZ) injection and the rats were examined 4 (4WDM) and 20 weeks (20WDM) after the treatment. Ischemia and reperfusion were induced by occlusion and reperfusion (I/R) of the left coronary artery (LCA). I/R-induced infarct size was determined using triphenyltetrazolium chloride (TTC) staining. After 20 weeks of STZ treatment (20WDM+I/R), the infarct size in the rat heart increased by 65.2 +/- 4.3%, whereas after 4 weeks of STZ treatment (4WDM+I/R), the infarct size decreased compared with the time-matched I/R group (43.1 +/- 3.6% and 59.5 +/- 5.6%, respectively). The number of dead myocytes including necrotic and apoptotic cells was determined using horseradish peroxidase (HRP) and terminal deoxynucleotide nick-end labeling (TUNEL) methods. The number of dead myocytes decreased in the 4WDM+I/R group, while the number of dead myocytes increased markedly in the 20WDM+I/R group, compared with the time-matched I/R group. The increment of ERK1/2 phosphorylation in the 4WDM group and the slight enhancement of this phosphorylation by I/R treatment were observed by western blotting. However, in the 20WDM group, the level of ERK1/2 phosphorylation reduced by approximately 1/3 compared with the time-matched control group; moreover, I/R treatment did not enhance the phosphorylation level. This study demonstrated that short- and long-term hyperglycemias exert opposite influences on ischemic myocardial injury, and these contradictory influences may depend on an ERK1/2 mediated intracellular signaling pathway. PMID- 14738871 TI - Fluvastatin prevents nephropathy likely through suppression of connective tissue growth factor-mediated extracellular matrix accumulation. AB - Diabetic nephropathy is related to glomerular extracellular matrix (ECM) accumulation that leads to glomerulosclerosis. Fluvastatin as a lipid-lowering medicine significantly prevents diabetic nephropathy, probably not only through its lipid-lowering action, but also mainly through its direct suppression of glomerular ECM accumulation. To test this hypothesis, in the present study, a five-sixths nephrectomized (5/6Nx) rat model to induce a renal ECM accumulation without coexistence of hyperlipidemia was used to investigate the effect of fluvastatin on renal function, glomerular ECM accumulation and expression of connective tissue growth factor (CTGF). 5/6Nx induced a significant nephropathy in rats at 13 weeks, indicated by renal dysfunction including increases in blood urine nitrogen, creatinine and urinary protein excretion, and renal histopathological changes. Administration of fluvastatin significantly prevented the renal dysfunction and histological abnormalities in the 5/6Nx rats. Furthermore, both significant suppression of matrix metalloproteinases (MMPs) activity such as MMP-2 and significant activation of tissue inhibitors of MMP (TIMPs) such as TIMP-2 observed in the 5/6Nx rats were almost completely prevented by fluvastatin, resulting in a significant prevention of glomerular ECM accumulation. For upstream mediator of ECM accumulation, 5/6Nx significantly up regulated CTGF mRNA expression, but fluvastatin treatment prevented CTGF up regulation. These results suggest that fluvastatin, as one of well-known lipid lowering agents, plays an important role in the prevention of nephropathy, likely through suppression of CTGF-mediated ECM accumulation. Therefore, fluvastatin may be a potential candidate for developing a pharmaceutical approach to the prevention of diabetic nephropathy due to its both lipid-lowering and direct anti renal ECM accumulation actions. PMID- 14738872 TI - Systemic up-regulation of sTNFR2 and IL-6 in Porphyromonas gingivalis pneumonia in mice. AB - Aspiration pneumonia is a common cause of death in older people, and the pathophysiology is a chronic respiratory failure with a mild airway inflammation. In this study, we established a mild inflammatory pneumonia model using Porphyromonas gingivalis (Pg) pathogen-infected mice. It elucidated the effects of Pg-infected pneumonia on proinflammatory cytokines tumor necrosis factor (TNF) alpha, interleukin-6 (IL-6), and IL-1beta production in both lung tissue and serum. We also elucidated production of soluble (s) TNF receptor (R) s, because TNF-alpha is considered to be a dominant inflammatory mediator. Lung TNF-alpha levels significantly increased at 2 h after infection, and rapidly returned to basal level at 24 h. Consistent with increase of TNF-alpha, remarkable increase of sTNFR2 but not sTNFR1 was detected in lung tissue from 2 to 72 h. Interestingly, sTNFR2/sTNFR1 ratio was significantly enhanced at 2 h in serum. In addition, lung IL-1beta and IL-6 levels also significantly increased from 2 to 24 h. Importantly, we found that IL-6 levels in serum reflected its local level. These results may suggest that systemically produced sTNFR2 and IL-6 could be a key role to modulate proinflammatory activities of TNF-alpha in Pg-induced lung inflammation simulated aspiration pneumonia. PMID- 14738873 TI - Heparin-binding EGF-like growth factor regulates human extravillous cytotrophoblast development during conversion to the invasive phenotype. AB - Cytotrophoblasts of the anchoring villi convert during human placentation from a transporting epithelium to an invasive, extravillous phenotype that expresses a distinct repertoire of adhesion molecules. Developing extravillous trophoblasts accumulate heparin-binding EGF-like growth factor (HB-EGF), a multifunctional cytokine, which binds HER1 and HER4 of the human EGF receptor (HER/ErbB) family. HB-EGF is downregulated in placentae of women with preeclampsia, a disorder associated with deficient trophoblast invasion, raising important questions about its physiological impact on cytotrophoblasts. Addition of HB-EGF during explant culture of first-trimester chorionic villi enhanced extravillous trophoblast differentiation and invasive activity. Using a first-trimester human cytotrophoblast line, the potential for autocrine and paracrine regulation of the developing trophoblast was established based on the expression of all four HER isoforms, as well as HB-EGF and related growth factors. HB-EGF did not alter proliferation, but initiated extravillous differentiation, with decreased alpha6 integrin expression, increased alpha1, and elevated cell migration. Function blocking antibodies against EGF family members reduced basal cell motility and antibody inhibition of either HER1 or HER4 ligation prevented HB-EGF-induced integrin switching. We conclude that HER-mediated autocrine and paracrine signaling by HB-EGF or other EGF family members induces cytotrophoblast differentiation to an invasive phenotype. PMID- 14738874 TI - The nuclear orphan receptor COUP-TFI is important for differentiation of oligodendrocytes. AB - We report here that a member of the nuclear hormone receptor superfamily, chicken ovalbumin upstream promoter-transcription factor 1 (COUP-TFI), plays a critical role in glial cell development and subsequent central nervous system myelination. We demonstrate that COUP-TF1 is expressed in cells of oligodendrocyte lineage. Furthermore, we demonstrate that COUP-TFI null mutant mice exhibit delayed axon myelination and increased dysmyelination in the central nervous system. Using in vitro differentiation assays, we show that these myelination defects are due to delays in oligodendrocyte differentiation. Finally, in situ hybridization and transfection analysis suggests that COUP-TFI acts as an upstream regulator of SCIP/Oct-6/Tst-1, a transcription factor involved in axon myelination. Taken together, these results suggest that COUP-TFI is an important regulator of oligodendrocyte differentiation. PMID- 14738875 TI - Combined intrinsic and extrinsic influences pattern cranial neural crest migration and pharyngeal arch morphogenesis in axolotl. AB - Cranial neural crest cells migrate in a precisely segmented manner to form cranial ganglia, facial skeleton and other derivatives. Here, we investigate the mechanisms underlying this patterning in the axolotl embryo using a combination of tissue culture, molecular markers, scanning electron microscopy and vital dye analysis. In vitro experiments reveal an intrinsic component to segmental migration; neural crest cells from the hindbrain segregate into distinct streams even in the absence of neighboring tissue. In vivo, separation between neural crest streams is further reinforced by tight juxtapositions that arise during early migration between epidermis and neural tube, mesoderm and endoderm. The neural crest streams are dense and compact, with the cells migrating under the epidermis and outside the paraxial and branchial arch mesoderm with which they do not mix. After entering the branchial arches, neural crest cells conduct an "outside-in" movement, which subsequently brings them medially around the arch core such that they gradually ensheath the arch mesoderm in a manner that has been hypothesized but not proven in zebrafish. This study, which represents the most comprehensive analysis of cranial neural crest migratory pathways in any vertebrate, suggests a dual process for patterning the cranial neural crest. Together with an intrinsic tendency to form separate streams, neural crest cells are further constrained into channels by close tissue apposition and sorting out from neighboring tissues. PMID- 14738876 TI - Xvelo1 uses a novel 75-nucleotide signal sequence that drives vegetal localization along the late pathway in Xenopus oocytes. AB - Vegetally localized RNAs in Xenopus laevis oocytes are involved in the patterning of the early embryo as well as in cell fate specification. Here we report on the isolation and characterization of a novel, vegetally localized RNA in Xenopus oocytes termed Xvelo1. It encodes a protein of unknown biological function and it represents an antisense RNA for XPc1 over a length of more than 1.8 kb. Xvelo1 exhibits a localization pattern reminiscent of the late pathway RNAs Vg1 and VegT; it contains RNA localization elements (LE) which do not match with the consensus structural features as deduced from Vg1 and VegT LEs. Nevertheless, the protein binding pattern as observed for Xvelo1-LE in UV cross-linking experiments and coimmunoprecipitation assays is largely overlapping with the one obtained for Vg1-LE. These observations suggest that the structural features recognized by the protein machinery that drives localization of maternal mRNAs along the late pathway in Xenopus oocytes must be redefined. PMID- 14738877 TI - TGF-beta superfamily members modulate growth, branching, shaping, and patterning of the ureteric bud. AB - Protein-rich fractions inhibitory for isolated ureteric bud (UB) growth were separated from a conditioned medium secreted by cells derived from the metanephric mesenchyme (MM). Elution profiles and immunoblotting indicated the presence of members of the transforming growth factor-beta (TGF-beta) superfamily. Treatment of cultured whole embryonic kidney with BMP2, BMP4, activin, or TGF-beta1 leads to statistically significant differences in the overall size of the kidney, the number of UB branches, the length and angle of the branches, as well as in the thickness of the UB stalks. Thus, the pattern of the ureteric tree is altered. LIF, however, appeared to have only minimal effect on growth and development of the whole embryonic kidney in organ culture. The factors all directly inhibited, in a concentration-dependent fashion, the growth and branching of the isolated UB, albeit to different extents. Antagonists of some of these factors reduced their inhibitory effect. Detailed examination of TGF-beta1-treated UBs revealed only a slight increase in the amount of apoptosis in tips by TUNEL staining, but diminished proliferation throughout by Ki67 staining. These data suggest an important direct modulatory role for BMP2, BMP4, LIF, TGF-beta1, and activin (as well as their antagonists) on growth and branching of the UB, possibly in shaping the growing UB by playing a role in determining the number of branches, as well as where and how the branches occur. In support of this notion, UBs cultured in the presence of fibroblast growth factor 7 (FGF7), which induces the formation of globular structures with little distinction between the stalk and ampullae [Mech. Dev. 109 (2001) 123], and TGF beta superfamily members lead to the formation of UBs with clear stalks and ampullae. This indicates that positive (i.e., growth and branch promoting) and negative (i.e., growth and branch inhibiting) modulators of UB morphogenesis can cooperate in the formation of slender arborized UB structures similar to those observed in the intact developing kidney or in whole embryonic kidney organ culture. Finally, purification data also indicate the presence of an as yet unidentified soluble non-heparin-binding activity modulating UB growth and branching. The data suggest how contributions of positive and negative growth factors can together (perhaps as local bipolar morphogenetic gradients existing within the mesenchyme) modulate the vectoral arborization pattern of the UB and shape branches as they develop, thereby regulating both nephron number and tubule/duct caliber. We suggest that TGF-beta-like molecules and other non heparin-binding inhibitory factors can, in the appropriate matrix context, facilitate "braking" of the branching program as the UB shifts from a rapid branching stage (governed by a feed-forward mechanism) to a stage where branching slows down (negative feedback) and eventually stops. PMID- 14738878 TI - Male infertility due to germ cell apoptosis in mice lacking the thiamin carrier, Tht1. A new insight into the critical role of thiamin in spermatogenesis. AB - A mouse model of thiamin-responsive megaloblastic anemia (diabetes mellitus, deafness, megaloblastic anemia) lacking functional Slc19a2 has been generated and unexpectedly found to have a male-specific sterility phenotype. We describe here the characterization of the testis-specific effects of absence of the high affinity thiamin transporter, Tht1. Null males were found to have hypoplastic testes secondary to germ cell depletion. Morphologic and expression analysis revealed that under conditions of standard thiamin intake, tissues affected in the syndrome (pancreatic beta-cell, hematopoietic cells, auditory nerve) maintained normal function but pachytene stage spermatocytes underwent apoptosis. Under conditions of thiamin challenge, the apoptotic cell loss extended to earlier stages of germ cells but spared Sertoli cells and Leydig cells. Injection of high-dose thiamin was effective in reversing the spermatogenic failure, suggesting that the absence of the thiamin carrier could be overcome by diffusion mediated transport at supranormal thiamin concentrations. These observations demonstrated that male germ cells, particularly those with high thiamin transporter expression beyond the blood-testis barrier, were more susceptible to apoptosis triggered by intracellular thiamin deficiency than any other tissue type. The findings described here highlight an unexpected and critical role for thiamin transport and metabolism in spermatogenesis. PMID- 14738879 TI - Alpha-endosulfine, a potential regulator of insulin secretion, is required for adult tissue growth control in Drosophila. AB - Alpha-endosulfine is a small protein that has been proposed to regulate ion channel activity and insulin secretion, but in vivo studies have been lacking. We have previously established the Drosophila ovary as a model system in which to study adult tissue growth regulation, and demonstrated a role of the insulin pathway in the proliferative response of ovarian cells to nutritional changes. Here, we find that the Drosophila alpha-endosulfine (dendos) gene, whose protein is expressed in germline and somatic cells of the ovary, as well as in the brain and certain regions of the intestine, is also required for this response. This requirement is non-cell autonomous, which is consistent with a role of dendos in secretion of Drosophila insulin-like peptides (DILPs), required for the proliferative response to nutritional changes. Our results show that dendos is also required for a distinct process in oogenesis, namely, the osmotic regulation of stage 14 oocytes, and that this requirement is cell autonomous, consistent with the role in ion channel regulation suggested by studies of the mammalian homologues. PMID- 14738880 TI - Pristionchus pacificus vulva formation: polarized division, cell migration, cell fusion, and evolution of invagination. AB - Tube formation is a widespread process during organogenesis. Specific cellular behaviors participate in the invagination of epithelial monolayers that form tubes. However, little is known about the evolutionary mechanisms of cell assembly into tubes during development. In Caenorhabditis elegans, the detailed step-to-step process of vulva formation has been studied in wild type and in several mutants. Here we show that cellular processes during vulva development, which involve toroidal cell formation and stacking of rings, are conserved between C. elegans and Pristionchus pacificus, two species of nematodes that diverged approximately 100 million years ago. These cellular behaviors are divided into phases of cell proliferation, short-range migration, and cell fusion that are temporally distinct in C. elegans but not in P. pacificus. Thus, we identify heterochronic changes in the cellular events of vulva development between these two species. We find that alterations in the division axes of two equivalent vulval cells from Left-Right cleavage in C. elegans to Anterior Posterior division in P. pacificus can cause the formation of an additional eighth ring. Thus, orthogonal changes in cell division axes with alterations in the number and sequence of cell fusion events result in dramatic differences in vulval shape and in the number of rings in the species studied. Our characterization of vulva formation in P. pacificus compared to C. elegans provides an evolutionary-developmental foundation for molecular genetic analyses of organogenesis in different species within the phylum Nematoda. PMID- 14738881 TI - Expression of activin subunits and receptors in the developing human ovary: activin A promotes germ cell survival and proliferation before primordial follicle formation. AB - The formation of the essential functional unit of the ovary, the primordial follicle, occurs during fetal life in humans. Factors regulating oogonial proliferation and interaction with somatic cells before primordial follicle formation are largely unknown. We have investigated the expression, localisation and functional effects of activin and its receptors in the human fetal ovary at 14-21 weeks gestation. Expression of mRNA for the activin betaA and betaB subunits and the activin receptors ActRIIA and ActRIIB was demonstrated by RT PCR. Expression of betaA mRNA increased 2-fold across the gestational range examined. Activin subunits and receptors were localised by immunohistochemistry. The betaA subunit was expressed by oogonia, and the betaB subunit and activin receptors were expressed by both oogonia and somatic cells. BetaA expression was increased in larger oogonia at later gestations, but was low in oocytes within newly formed primordial follicles. Treatment of ovary fragments with activin A in vitro increased both the number of oogonia present and oogonial proliferation, as detected by bromodeoxyuridine (BrdU) incorporation. These data indicate that activin may be involved in the autocrine and paracrine regulation of germ cell proliferation in the human ovary during the crucial period of development leading up to primordial follicle formation. PMID- 14738882 TI - DSCR1 gene expression is dependent on NFATc1 during cardiac valve formation and colocalizes with anomalous organ development in trisomy 16 mice. AB - The Down syndrome critical region 1 (DSCR1) gene is present in the region of human chromosome 21 and the syntenic region of mouse chromosome 16, trisomy of which is associated with congenital heart defects observed in Down syndrome. DSCR1 encodes a regulatory protein in the calcineurin/NFAT signal transduction pathway. During valvuloseptal development in the heart, DSCR1 is expressed in the endocardium of the developing atrioventricular and semilunar valves, the muscular interventricular septum, and the ventricular myocardium. Human DSCR1 contains an NFAT-rich calcineurin-responsive element adjacent to exon 4. Transgenic mice generated with a homologous regulatory region of the mouse DSCR1 gene linked to lacZ (DSCR1(e4)/lacZ) show gene activation in the endocardium of the developing valves and aorticopulmonary septum of the heart, recapitulating a specific subdomain of endogenous DSCR1 cardiac expression. DSCR1(e4)/lacZ expression in the developing valve endocardium colocalizes with NFATc1 and, endocardial DSCR1(e4)/lacZ, is notably reduced or absent in NFATc1(-/-) embryos. Furthermore, expression of the endogenous DSCR1(e4) isoform is decreased in the outflow tract of NFATc1(-/-) hearts, and the DSCR1(e4) intragenic element is trans-activated by NFATc1 in cell culture. In trisomy 16 (Ts16) mice, expression of endogenous DSCR1 and DSCR1(e4)/lacZ colocalizes with anomalous valvuloseptal development, and transgenic Ts16 hearts have increased beta-galactosidase activity. DSCR1 and DSCR1(e4)/lacZ also are expressed in other organ systems affected by trisomy 16 in mice or trisomy 21 in humans including the brain, eye, ear, face, and limbs. Together, these results show that DSCR1(e4) expression in the developing valve endocardium is dependent on NFATc1 and support a role for DSCR1 in normal cardiac valvuloseptal formation as well as the abnormal development of several organ systems affected in individuals with Down syndrome. PMID- 14738883 TI - Roles of erbB4, rhombomere-specific, and rhombomere-independent cues in maintaining neural crest-free zones in the embryonic head. AB - Within the developing vertebrate head, the migration of neural tube-derived neural crest cells (NCCs) through the cranial mesenchyme is patterned into three streams, with mesenchyme adjacent to rhombomeres (r)3 and r5 maintained NCC-free. The receptor tyrosine kinase erbB4 is expressed within r3 and r5 and is required to maintain the r3-adjacent NCC-free zone in mouse embryos. In this study, we demonstrate that the extent of r3 involvement in patterning mouse NCC migration is restricted to the same dorsolateral region regulated by erbB4. In chick embryos, we show that erbB4 signaling similarly maintains the r3-adjacent NCC free zone. However, although r5 expresses erbB4, this is insufficient to maintain the r3-adjacent NCC-free zone in grafting experiments where r5 replaced r3, indicating that erbB4 requires additional factors at the A-P level of r3 to pattern NCC migration. Furthermore, we show that the r5-adjacent NCC-free zone is maintained independently of r5, but requires surface ectoderm. Finally, we demonstrate that avian cranial surface ectoderm is patterned molecularly, with dorsolateral surface ectoderm at the levels of r2/3 and r7 expressing the sulfatase QSulf1 in quail, or the orthologue CSulf1 in chick. Aberrant NCC migration into r3-adjacent mesenchyme correlated with more focused QSulf1 expression in r2/3 surface ectoderm. PMID- 14738884 TI - Transcriptional control of glial and blood cell development in Drosophila: cis regulatory elements of glial cells missing. AB - In Drosophila, glial cell differentiation requires the expression of glial cells missing (gcm) in multiple neural cell lineages, where gcm acts as a binary switch for glial vs. neuronal fate. Thus, the primary event controlling gliogenesis in neural progenitors is the transcription of gcm. In addition, gcm is also required for the differentiation of macrophages, and is expressed in the hemocyte lineage. This dual role of gcm in glial cell and blood cell development underscores the need for the precise temporal and spatial regulation of gcm transcription. To understand how gcm transcription is regulated, we have undertaken an analysis of the cis-regulatory DNA elements of gcm using lacZ reporter activity in transgenic embryos, testing the activity of approximately 35 kilobases of DNA from the gcm locus. We have identified several distinct DNA regions that promote most of the elements of gcm expression. These include elements for general neural expression, gcm-independent and gcm-dependent glial-specific expression, as well as early and late hemocyte expression. We show that expression of an abdominal glial-specific element is dependent on the homeotic gene abdominal-A. Our results indicate that gcm transcription is controlled by a combination of general and lineage-specific elements, positive autoregulation, and neuronal repression. PMID- 14738885 TI - An early pharyngeal muscle enhancer from the Caenorhabditis elegans ceh-22 gene is targeted by the Forkhead factor PHA-4. AB - Caenorhabditis elegans pharyngeal muscle development involves ceh-22, an NK-2 family homeobox gene related to genes controlling heart development in other species. ceh-22 is the earliest known gene expressed in the pharyngeal muscles and is likely regulated directly by factors specifying pharyngeal muscle fate. We have previously implicated the ceh-22 distal enhancer in initiating ceh-22 expression. Here we analyze the distal enhancer using functional and comparative assays. The distal enhancer contains three subelements contributing additively to its activity, and functionally important regulatory sequences are highly conserved in Caenorhabditis briggsae. One subelement, termed DE3, is strongly active in the pharyngeal muscles, and we identified two short oligonucleotides (de199 and de209) contributing to DE3 activity. Multimerized de209 enhances transcription similarly to DE3 specifically in the pharyngeal muscles, suggesting it may be an essential site regulating ceh-22. de209 binds the pan-pharyngeal Forkhead factor PHA-4 in vitro and responds to ectopic pha-4 expression in vivo, suggesting that PHA-4 directly initiates ceh-22 expression through de209. Because de209 enhancer activity is primarily limited to the pharyngeal muscles, we hypothesize that de209 also binds factors functioning with PHA-4 to specifically activate ceh-22 expression in pharyngeal muscle. PMID- 14738886 TI - Expression and function of conserved nuclear receptor genes in Caenorhabditis elegans. AB - The Caenorhabditis elegans genome encodes 284 nuclear receptor (NR) genes. Among these 284 NR genes are 15 genes conserved among the Metazoa. Here, we analyze the expression and function of eight heretofore uncharacterized conserved C. elegans NR genes. Reporter gene analysis demonstrates that these genes have distinct expression patterns and that a majority of the C. elegans cell types express a conserved NR gene. RNA interference with NR gene function resulted in visible phenotypes for three of the genes, revealing functions in various processes during postembryonic development. Five of the conserved NR genes are orthologs of NR genes that function during molting and metamorphosis in insects. Functional studies confirm a role for most of these 'ecdysone cascade' NR orthologs during the continuous growth and dauer molts. Transcript levels for these genes fluctuate in a reiterated pattern during the molting cycles, reminiscent of the expression hierarchy observed in the insect ecdysone response. Together, these analyses provide a foundation for further dissecting the role of NRs in nematode development as well as for evaluating conservation of NR functions among the Metazoa. PMID- 14738887 TI - Toll-like receptor-2 is essential in murine defenses against Candida albicans infections. AB - In this work, we studied the role of toll-like receptor-2 (TLR2) in murine defenses against Candida albicans. TLR2-deficient mice experimentally infected intraperitoneally (i.p.) or intravenously (i.v.) in vivo had very significant impaired survival compared with that of control mice. In vitro production of TNF alpha and macrophage inhibitory protein-2 (MIP-2) by macrophages from TLR2-/- mice in response to yeasts and hyphae of C. albicans were significantly lower (80% and 40%, respectively; P <0.05) than production by macrophages from wild type mice. This impaired production of TNF-alpha and MIP-2 probably contributed to the 41% decreased recruitment of neutrophils to the peritoneal cavity of i.p. infected TLR2-/- mice. In contrast, in vitro phagocytosis of yeasts and production of reactive oxygen intermediates (ROI) were not affected in macrophages from TLR2-/- animals. Our data indicate that TLR2 plays a major role in the response of macrophages to C. albicans, triggering cytokine and chemokine expression, and it is essential for in vivo protection against infection. PMID- 14738888 TI - Antigen-specific CD8+ T cell responses in intestinal tissues during murine listeriosis. AB - Infection of mice with Listeria monocytogenes induces a strong CD8+ T cell response, which is critical for the control of bacteria and for protection against re-infection. We analyzed the CD8+ T cell response in different intestinal tissues following oral and intravenous (i.v.) L. monocytogenes infection. After oral infection, bacterial titers in small intestine and large intestine, and the listeria-specific CD8+ T cell response in the mucosa of both parts of the intestine, were highly correlated. Oral infection of CD28-deficient mice revealed that this response was strictly dependent on CD28 costimulation. Significant listeria-specific CD8+ T cell responses also occurred in all intestinal tissues analyzed after i.v. infection or after DNA vaccination, indicating that the accumulation of listeria-specific CD8+ T cells in these tissues only partially depends on local antigen presentation and inflammation. PMID- 14738889 TI - Cytomegalovirus infection aggravates atherogenesis in apoE knockout mice by both local and systemic immune activation. AB - Since the 1970s, cytomegalovirus (CMV) infection has been associated with atherosclerotic disease. However, the exact contribution of the virus remains uncertain. In this article we describe both a direct and indirect immune-mediated effect of the virus on the disease process. Eight-week-old apolipoprotein E (apoE) knockout mice were infected with mouse CMV (MCMV) or mock injected, and they were sacrificed at 2 and 20 weeks post-injection (p.i.) to study atherosclerosis, vascular wall IFNgamma and TNFalpha expression and MCMV spread. To study plasma IFNgamma and TNFalpha levels, blood was collected at 1, 2, 4 and 6 days p.i. in addition to days of sacrifice. Plasma cytokine levels were increased after MCMV infection at early time points and decreased to mock levels at 2 and 20 weeks p.i. At 2 weeks p.i., more aortic arch samples showed local cytokine expression after MCMV infection. The number of early atherosclerotic lesions and the percentage of mice containing early lesions were increased at 2 weeks p.i., while at 20 weeks p.i., the MCMV-induced effect on atherogenesis was seen on the late lesions. In conclusion, MCMV infection induces a systemic immune response reflecting an indirect effect of MCMV infection on atherosclerosis in addition to a local aortic immune response reflecting a direct effect of the virus on the atherosclerotic process. PMID- 14738890 TI - Monocyte cytokine secretion in patients with pulmonary tuberculosis differs from that of healthy infected subjects and correlates with clinical manifestations. AB - Cell-mediated immunity, leading to Mycobacterium tuberculosis (Mtb)-constraining granuloma formation, is the major component of host defense against tuberculosis and is regulated by the balance of cytokines secreted mostly by mononuclear phagocytes and lymphocytes. To better understand the role of monocytes in the regulation of the immune response against pulmonary tuberculosis, we examined IL 10, IL-12 and TNF-alpha release by monocytes from healthy purified protein derivative (PPD) reactors and pulmonary tuberculosis patients with or without systemic reactions (e.g., fever, weight loss, asthenia). Our study shows that, probably as a result of in vivo priming by circulating antigens, monocytes from patients, especially those with systemic manifestations, have a biased ex vivo cytokine secretion, with high IL-10 and TNF-alpha but low IL-12, in contrast with PPD reactors. Higher spontaneous IL-10 and TNF-alpha release persisted when monocytes were co-cultured with autologous lymphocytes. Challenge of patients' monocytes with a virulent Mtb strain led to a further enhancement of IL-10 and TNF-alpha, but not of IL-12. When lymphocytes were added to these cultures, IL-10 and TNF-alpha elevation persisted and, in the patients with a systemic reaction, both IL-12 and IFN-gamma were significantly reduced compared to PPD reactors. Intragroup comparisons revealed that in the patients with systemic reactions, the lymphocyte-monocyte interaction resulted in a positive feedback for IL-10 secretion, while in the patients without systemic reaction and PPD reactors, the feedback was positive for IL-12 secretion. Thus, in tuberculosis, there appears to exist a relationship between the immunological findings and the distinct clinical manifestations. PMID- 14738891 TI - Effects of quorum-sensing on immunoglobulin G responses in a rat model of chronic lung infection with Pseudomonas aeruginosa. AB - Levels of serum antibodies against Pseudomonas aeruginosa were observed for 106 days in a rat model of chronic lung infection. Significantly weaker responses of serum IgG and IgG1 and a lower ratio of IgG1/IgG2a were found in the rats infected with the quorum-signal-deficient mutant, PAO1 (rhlI, lasI), compared with the wild-type PAO1. Four out of 15 rats infected with wild-type PAO1 contained bacteria in the lungs on day 106, whereas no bacteria were found in the mutant PAO1 group. The results indicate that quorum signals contribute to the persistence of the infection and influence the immune response. PMID- 14738892 TI - Enteropathogenic Escherichia coli outer membrane proteins induce changes in cadherin junctions of Caco-2 cells through activation of PKCalpha. AB - Enteropathogenic Escherichia coli (EPEC) is a Gram-negative bacterial pathogen that adheres to human intestinal epithelial cells, resulting in watery, persistent diarrhoea. Despite the advances made in understanding EPEC-host cell interactions, the molecular mechanisms underlying watery diarrhoea have not been understood fully. Loss of transepithelial resistance and increased monolayer permeability by disruption of tight junctions has been implicated in this process. Apart from disruption of tight junctions, an important factor known to regulate monolayer permeability is E-cadherin and its interaction with beta catenin, both of which constitute the adherens junctions. Our previous studies using HEp-2 cells demonstrated the morphological and cytoskeletal changes caused by cell-free outer membrane preparations (OMPs) of EPEC. In this study, we have shown that EPEC and its OMP induce significant changes in the adherens junctions of Caco-2 monolayers. We also observed significant phosphorylation of protein kinase Calpha (PKCalpha) in cells treated with either whole EPEC or its OMP. Immunoprecipitation of cell lysates with anti-E-cadherin and probing with phospho PKCalpha monoclonal antibodies and anti-beta-catenins revealed that in these cells, phosphorylated PKCalpha is associated with cadherins, leading to the dissociation of the cadherin/beta-catenin complex. Immunofluorescence showed beta catenins dissociated from the membrane-bound cadherins and redistributed into the cytoplasm. Expression of dominant negative PKCalpha reversed these effects caused by either whole EPEC or its OMP and also reduced the associated increase in monolayer permeability. It is possible that this mechanism may complement the earlier known pathways for loss of barrier function involving myosin light chain kinase activation and also may play a role in causing host cell death by apoptosis. PMID- 14738893 TI - Hepatocyte growth factor inhibits intrinsic antibacterial activity of Madin-Darby canine kidney cells. AB - We investigated whether or not polarized renal epithelial cells produce antibacterial factors, which aid in host defense at the cell surface of renal epithelium. A model of polarized Madin-Darby canine kidney (MDCK) epithelial cells grown on filters was used to test for the presence of apically or basolaterally secreted factors on the growth of non-virulent (XL1-Blue) and uropathogenic (J96) strains of Escherichia coli (E. coli). Growth of both XL1 Blue and J96 strains of E. coli in medium on the apical and basolateral surface of MDCK cells was inhibited as compared to bacterial growth in medium not exposed to MDCK cells. The inhibition of bacterial growth was similar in both apical and basolateral surface medium. Pretreatment of MDCK cells with hepatocyte growth factor (HGF) blunted the inhibition of XL1-Blue and J96 growth in apical and basolateral surface medium as compared to growth in medium on the surfaces of untreated MDCK cells. Immunofluorescent analysis demonstrated the presence of beta-defensin isoforms 1-3 in MDCK cells, with isoform 1 being the most prevalent form observed. HGF treatment reduced the amount of immunoreactive beta-defensin-1 in MDCK cells. These data demonstrate that polarized renal epithelium produce antibacterial factors. The renotropic growth factor HGF inhibits these antibacterial factors. beta-defensins may contribute to this antibacterial activity and play an important role in renal epithelial resistance to bacterial infections. PMID- 14738894 TI - Characterization of two lipoproteins in Pasteurella multocida. AB - An in vivo expression technology (IVET) system was previously developed and used to identify Pasteurella multocida genes, which are upregulated during infection of the host. Of the many genes identified, two encoded products which showed similarity to the Haemophilus influenzae lipoproteins, protein D and PCP, which have been shown to stimulate heterologous immunity against infection with H. influenzae. Therefore, the lipoprotein homologues in P. multocida, designated GlpQ and PCP, were investigated. GlpQ and PCP were shown to be lipoproteins by demonstrating that post-translational processing of the proteins was inhibited by globomycin. The P. multocida GlpQ homologue showed glycerophosphodiester phosphodiesterase enzyme activity, indicating that it is a functional homologue of other characterized GlpQ enzymes. Using surface immunoprecipitation, PCP was found to be surface exposed, but GlpQ was not. Non-lipidated forms of GlpQ and PCP were expressed and purified from Escherichia coli and used to vaccinate mice. However, mice were not protected from challenge with live P. multocida. The lipoproteins were then expressed in E. coli in the lipidated form and used to vaccinate mice and chickens. Protection against challenge with live P. multocida was not observed. PMID- 14738895 TI - Role of environment and behaviour in familial resemblances of Plasmodium falciparum infection in a population of Senegalese children. AB - Despite the importance of both environment and behaviour in vector-borne disease epidemiology, these factors are unable to explain alone the distribution of cases in a community and the diversity of clinical presentations, suggesting the involvement of more individual factors such as age, sex, immunity or genetic background. The existence of a genetic factor involved in the susceptibility/resistance to a disease can be suspected by the demonstration of a familial aggregation of cases or by the stability over time of infectious status (infected vs. uninfected; mean level of parasite density (PD), etc.). These familial resemblances can be explained by shared environment, family habits and behaviours (use of bed nets, field activities, etc.). In this preliminary study, we essentially investigated the influence of environment and behaviour on Plasmodium falciparum infection levels and reported the effects of these factors on the existence of familial resemblances. Our results are consistent with the existence of familial resemblances for both the level of P. falciparum infection and the qualitative infection status (QIS) (infected vs. uninfected) that seem to be more related to shared behaviour and environment than to a genetic factor. However, although familial resemblances decreased significantly when adjusted for shared behaviour and environment, this decrease is around 12% for the variability between families, against only 4.5% of that within families. Furthermore, we also demonstrated that the QIS is remarkably stable over time. Both these results are consistent with the hypothesis of the existence of a strong and complex individual factor involved in the control of infection status. PMID- 14738896 TI - Expression of small hairpin RNA by lentivirus-based vector confers efficient and stable gene-suppression of HIV-1 on human cells including primary non-dividing cells. AB - RNA interference (RNAi) is a sequence-specific RNA degradation process mediated by short double-stranded RNAs termed small interfering RNAs. Here, we describe the lentivirus-based vector small interfering RNA system expressing. As a pilot study, we generated constructs expressing small hairpin RNA (shRNA) specific for luciferase gene (shLuc) or green fluorescence protein (shGFP) under the control of human H1 RNA polymerase III promoter. The effect of the shRNA was evaluated against HIV-1 infection in a single-round or multiple-round infectious system using an HIV-1 molecular clone carrying the luc or GFP gene. In the single-round infectious system, cells transduced with shLuc by lentiviral vector significantly reduced (approximately 90% reduction) viral gene expression after challenge infection at a multiplicity of infection of 10. These transduced cells continued to resist against at least four sequentially repeated challenge infections. Importantly, this efficient antiviral activity persisted over 35 days in culture. In a multiple-round infectious system using a replication-competent HIV-1 molecular clone carrying the GFP gene, we also observed that a lentiviral vector expressing shGFP could inhibit HIV-1 replication for at least 1 week. The profound effect of lentiviral shRNA was also observed in human primary monocyte derived macrophages. Thus, shRNA introduced through the lentiviral vector can be useful for efficient and stable gene suppression in human cells including primary non-dividing cells. Moreover, quantitative analysis of viral cDNA synthesis on challenge infection showed that viral genomic RNAs packaged in incoming virus core might not be targeted by shLuc. Instead, the degradation of transcripts from integrated proviral DNAs might be a major cause of the profound reduction in HIV 1 gene expression by shRNA in our system. PMID- 14738897 TI - Phytoestrogen genistein as an anti-staphylococcal agent. AB - The soybean-derived isoflavone genistein has been shown to exert beneficial effects on many disorders, including cancer and cardiovascular diseases. The effects of genistein on mammalian cells are mediated by its abilities to inhibit topoisomerase II and protein tyrosine kinase. In order to examine the potential antibacterial activities of genistein, we incubated the bacteria with various concentrations of this compound for different periods of time and assessed the viable counts. Exposure to genistein exhibited an inhibitory effect on all staphylococcal strains tested, including methicillin-resistant strains. Furthermore, the growth of Streptococcus pasteurianus, Bacillus cereus, and Helicobacter pylori was clearly inhibited by genistein, whereas Escherichia coli growth was not suppressed. Daidzein, which is structurally similar to genistein, but deficient in topoisomerase II inhibitory activity, also inhibited the growth of Staphylococcus aureus, albeit with lower potency than genistein. Our results indicate that genistein exerts potent antibacterial properties in vitro, which are possibly mediated by the stabilization of the covalent topoisomerase II-DNA cleavage complex. PMID- 14738898 TI - Interaction between Brucella abortus and cellular prion protein in lipid raft microdomains. AB - A wide variety of pathogens employ lipid raft microdomains to infect host cells. Here, we review selected aspects of interaction between Brucella abortus and cellular prion protein, one of the lipid raft-associated molecules on the plasma membrane, when bacteria infect macrophages, and discuss the correlates of proliferation in mice. PMID- 14738899 TI - Adhesins and invasins of pathogenic bacteria: a structural view. AB - Adhesion and invasion of pathogenic bacteria represent the important initial step of infection. Pathogens utilize surface-located adhesins/invasins for specific interaction with host cell receptors. The three-dimensional structures of a number of adhesins/invasins show that many are elongated molecules containing domains commonly found in eukaryotic proteins. Similar folds are employed repeatedly to target different receptors. PMID- 14738900 TI - Pathogenesis and host responses in human onchocerciasis: impact of Onchocerca filariae and Wolbachia endobacteria. AB - Onchocerca volvulus is a tissue-invasive parasitic nematode causing skin and eye pathology in human onchocerciasis. The filariae habour abundant intracellular Wolbachia bacteria, now recognised as obligatory symbionts, and therefore emerging as a novel target for chemotherapy. Recent research demonstrates that both the filariae and endobacteria contribute to the pathogenesis of onchocerciasis, and molecules have been identified that promote inflammatory or counter-inflammatory immune mechanisms, divert the host's immune response or procure evasion of the parasite. PMID- 14738901 TI - Cell biology of the intracellular infection by Legionella pneumophila. AB - Legionella pneumophila has become a paradigm for facultative intracellular pathogens that modulate biogenesis of their phagosomes into replicative niches. The ability to alter host cell biology and tailor it into a hospitable host for intracellular proliferation is at the crux of the mechanism of pathogenesis of Legionnaires' disease. PMID- 14738902 TI - Age and monosodium glutamate treatment cause changes in the stimulation-induced [3H]-norepinephrine release from rat nucleus tractus solitarii-dorsal vagal nucleus slices. AB - In nucleus tractus solitarii-dorsal vagal nucleus slices prepared from young adult rats (180-260 g) 10(-3) M L-glutamate and 10(-5) M baclofen caused a 2-3 fold increase of field stimulation-induced [3H]-norepinephrine release without affecting the resting release. In slices prepared from rats treated neonatally with monosodium glutamate neither L-glutamate nor baclofen had any effect on stimulation-induced norepinephrine release, tested between postnatal days 74-99 (350-530 g). In untreated littermates used in the same period (460-580 g) L glutamate was fully effective whereas baclofen was ineffective. The tritium content in tissue extracts did not differ significantly in the three experimental groups. It is concluded that i) the loss of GABA(B) receptor-mediated disinhibitory stimulation of norepinephrine release is an age-related phenomenon and ii) neonatal monosodium glutamate treatment causes a damage in the local neural circuitry characterized by the loss of glutamate receptor-mediated mechanism that stimulates the release of norepinephrine. PMID- 14738903 TI - Dopaminergic ventral tegmental neurons modulated by methylphenidate. AB - Treatment of psychostimulants leads to the development of behavioral sensitization, an augmented behavioral response to drug re-administration. The induction of behavioral sensitization to psychostimulants such as amphetamine and cocaine occurs at the ventral tegmental area's dopaminergic neurons (VTA-DA). Currently, there is limited experimental data about the physiological properties of methylphenidate (MPD) on VTA-DA neurons. Behavioral and electrophysiological experiments using male rats were performed before and after MPD treatment. The behavioral experiment included dose-response (0.6, 2.5, and 10.0 mg/kg MPD) study to select the most effective dose for the electrophysiological study. Methylphenidate increased locomotion in typical dose response characteristics. Based on this experiment, the 10.0 mg/kg MPD was used in two types of electrophysiological recordings: 1) intracellular recording of neuronal activity performed on horizontal 275-300 microm brain slices and 2) whole-cell patch clamping before and after electrical stimulation to study post-synaptic currents on neurophysiologically identified VTA-DA neurons. Methylphenidate suppressed the neuronal activity of these neurons for 210 +/- 30 sec. Stimulation of the prefrontal cortex afferent fibers to these VTA-DA neurons in the presence of TTX, saclofen, and picrotoxin led to the conclusion that this input is mediated via NMDA and kainate/AMPA receptors and may participate to induce behavioral sensitization to psychostimulants. PMID- 14738904 TI - Nitric oxide of the supraoptic nucleus influences the salivary secretion, sodium renal excretion, urinary volume and arterial blood pressure induced by pilocarpine. AB - Male Holtzman rats weighting 200-250 g were anesthetized with zoletil 50 mg/Kg (tiletamine chloridrate 125.0 mg and zolazepan chloridrate 125.0 mg) into quadriceps muscle and stainless steel cannulas were implanted into their supraoptic nucleus (SON). We investigated the effects of the injection into the supraoptic nucleus (SON) of FK 409, a nitric oxide donor, and NW-nitro-L-arginine methyl ester (L-NAME), a nitric oxide synthase inhibitor (NOS), on the salivary secretion, arterial blood pressure, sodium excretion and urinary volume induced by pilocarpine, which was injected into SON. The drugs were injected in 0.5 microl volume over 30-60 s. Controls was injected with a similar volume of 0.15 M NaCl. FK 409 and L-NAME were injected at doses of 20 microg/0.5 microl and 40 microg/0.5 microl respectively. The amount of saliva secretion was studied over a five-minute period after injection of pilocarpine into SON. Injection of pilocarpine (10, 20, 40, 80, 160 microg/microl) into SON produced a dose dependent increase in salivary secretion. L-NAME was injected into SON prior to the injection of pilocarpine into SON, producing an increase in salivary secretion due to the effect of pilocarpine. FK 409 injected into SON attenuating the increase in salivary secretion induced by pilocarpine. Mean arterial pressure (MAP) increase after injections of pilocarpine into the SON. L-NAME injected into the SON prior to injection of pilocarpine into SON increased the MAP. FK 409 injected into the SON prior to pilocarpine attenuated the effect of pilocarpine on MAP. Pilocarpine (0.5 micromol/0.5 microl) injected into the SON induced an increase in sodium and urinary excretion. L-NAME injected prior to pilocarpine into the SON increased the urinary sodium excretion and urinary volume induced by pilocarpine. FK 409 injected prior to pilocarpine into the SON decreased the sodium excretion and urinary volume induced by pilocarpine. All these roles of pilocarpine depend on the release of nitric oxide into the SON. In summary the present results show: a) SON is involved in pilocarpine-induced salivation; b) that mechanism involves increase in MAP, sodium excretion and urinary volume. PMID- 14738905 TI - Postnatal development of hepatic oxidative, hydrolytic and conjugative drug metabolizing enzymes in female horses. AB - Little is known about the effects of aging on the hepatic drug metabolizing capacity of horses despite the relatively long lifespan characterizing this species. A wide array of cytochrome P450 (CYP)-dependent monooxygenases, carboxylesterases and transferases were assayed in liver microsomes from 50 female horses in an age range between less than 1 year to over 12 years. Rather unexpectedly, both the CYP content and the activity of NADPH cytochrome c reductase rose as a function of age. Accordingly, a general increasing trend was recorded in the rate of the in vitro metabolism of the substrates reported to be related to CYP2B-, CYP2E- or CYP3A, although, as detected by Western immunoblotting, only the levels of proteins recognized by anti-rat CYP3A- and CYP2B antibodies appeared to increase consistently. Also the carboxylesterases and uridindiphosphoglucuronyl-transferase (UGT) activity toward 1-naphthol displayed a similar trend, glutathione S-transferase accepting 3,4 dichloronitrobenzene as a substrate being the only enzyme activity showing an age related decline. A positive correlation was also found between liver cadmium content and CYP amount as well as the activities of most monooxygenases (except for those related to CYP1A), carboxylesterases, and UGT. While confirming that a number of enzyme activities are less expressed in foals, our results contradict the general view that the drug metabolizing capacity drops in elder individuals. Although several other factors can influence the kinetics of foreign compounds in aged animals, data from this study may provide insight in understanding possible age-related differences in drug efficacy and the response to toxic substances in horses. PMID- 14738906 TI - Naringin alters the cholesterol biosynthesis and antioxidant enzyme activities in LDL receptor-knockout mice under cholesterol fed condition. AB - The purpose of the current study was to evaluate the lipid lowering and antioxidant capacity of naringin in LDL receptor knockout (LDLR-KO) mice fed a cholesterol (0.1 g/100 g) diet. As such, naringin or lovastatin (0.02 g/100 g) was supplemented in a cholesterol diet for 6 weeks. The naringin and lovastatin supplementation significantly lowered the plasma total cholesterol level compared to the control group. The plasma and hepatic triglyceride level was only lowered by the lovastatin supplement, while the hepatic cholesterol content was lowered by both the naringin and lovastatin supplements compared to the control group. The hepatic HMG-CoA reductase activity was significantly lower in the naringin and lovastatin supplemented groups than in the control group, whereas the ACAT activity was unaffected. The excretion of total sterol was significantly higher in the naringin and lovastatin groups compared to the control group due to significant changes in the acidic and neutral sterol, respectively. When comparing the hepatic antioxidant enzyme activities, the superoxide dismutase, catalase, and glutathione reductase activities were all significantly higher in the naringin-supplemented group than in the control group, while only the lovastatin supplement increased the glutathione reductase activity. Accordingly, the current results confirmed that naringin lowers the plasma cholesterol level via the inhibition of hepatic HMG-CoA reductase activity and increases the excretion of fecal sterol. Naringin was also found to improve the activities of hepatic antioxidant enzymes against oxidative stress in a hypercholesterolemic animal model, i.e. cholesterol-fed LDLR-KO mice. PMID- 14738907 TI - Alteration in amino-glycerophospholipids levels in the plasma of children with autism: a potential biochemical diagnostic marker. AB - Currently, there is no biochemical test to assist in the behavioral diagnosis of autism. We observed that levels of phosphatidylethanolamine (PE) were decreased while phosphatidylserine (PS) were increased in the erythrocyte membranes of children with autism as compared to their non-autistic developmentally normal siblings. A new method using Trinitrobenezene sulfonic acid (TNBS) for the quantification of PE and PS (amino-glycerophospholipids, i.e., AGP) in the plasma of children was developed and standardized. Wavelength scans of TNBS-PE and TNBS PS complexes gave two peaks at 320 nm and 410 nm. When varying concentrations of PS and PE were used, a linear regression line was observed at 410 nm with TNBS. Using this assay, the levels of AGP were found to be significantly increased in the plasma of children with autism as compared to their non-autistic normal siblings. It is proposed that plasma AGP levels may function as a potential diagnostic marker for autism. PMID- 14738909 TI - Capacitative calcium entry in guinea pig gallbladder smooth muscle in vitro. AB - This study investigates the involvement of capacitative Ca2+ entry in excitation contraction coupling in guinea pig gallbladder smooth muscle. Thapsigargin (0.1 nM-1 microM, a sarcoplasmic reticulum Ca(2+)-ATPase inhibitor) produced slowly developing sustained tonic contractions in guinea pig isolated gallbladder strips. All contractions approached 50% of the response to carbachol (10 microM) after 55 min. Contractile responses to thapsigargin (1 microM) were abolished in a Ca(2+)-free medium. Subsequent re-addition of Ca2+ (2.5 mM) produced a sustained tonic contraction (99 +/- 6% of the carbachol response). The contractile response to Ca2+ re-addition following incubation of tissues in a Ca(2+)-free bathing solution in the absence of thapsigargin was significantly less than in its presence (79 +/- 4 % vs 100 +/- 7 % of carbachol; p < 0.05). Contractile responses to Ca2+ re-addition following treatment with thapsigargin were attenuated by (a) the L-type voltage-operated Ca2+ channel antagonist, nifedipine (10 microM) and (b) the general inhibitor of Ca2+ entry channels including store-operated channels, SK&F96365 (50 microM and 100 microM). In separate experiments, responses to Ca2+ re-addition were essentially abolished by the tyrosine kinase inhibitor, genistein (100 microM). These results suggest that capacitative Ca2+ entry provides a source of activator Ca2+ for guinea pig gallbladder smooth muscle contraction. Contractile responses to Ca2+ re-addition following depletion of sarcoplasmic reticulum Ca2+ stores with thapsigargin, are mediated in part by Ca2+ entry through voltage-operated Ca2+ channels and by capacitative Ca2+ entry through store-operated Ca2+ channels which can be blocked by SK&F96365. Furthermore, capacitative Ca2+ entry in this tissue may be modulated by tyrosine kinase. PMID- 14738908 TI - Antifibrotic effect of the Chinese herbs, Astragalus mongholicus and Angelica sinensis, in a rat model of chronic puromycin aminonucleoside nephrosis. AB - Nephrotic syndrome has long been treated in China with two herbs, Astragalus mongholicus and Angelica sinensis, which may have antifibrotic effects. METHODS: Rats with chronic puromycin-induced nephrosis were treated with Astragalus and Angelica 3 mL/d (n = 7) or enalapril 10 mg/kg/d (n = 7). Normal control rats (n = 7) received saline rather than puromycin, and an untreated control group (n = 7) received puromycin but no treatment. After 12 weeks, stained sections of the glomerulus and tubulointerstitium were evaluated for injury. Immunohistochemistry staining measured extracellular matrix components, transforming growth factor beta1 (TGFbeta1), osteopontin, ED-1-positive cells, and alpha-actin. TGFbeta1 mRNA was assessed by in situ hybridization. Renin, ACE activity, angiotensin, and aldosterone were measured by radioimmunoassay or colorimetry. In the untreated rats, chronic renal injury progressed to marked fibrosis at 12 weeks. Astragalus and Angelica significantly reduced deterioration of renal function and histologic damage. Expressions of type III and IV collagen, fibronectin, and laminin also decreased significantly. This anti-fibrotic effect was similar to that of enalapril. The herbs had no effect on the renin-angiotensin system but did reduce the number of ED-1-positive, and alpha-actin positive cells and expression of osteopontin compared to untreated controls. The combination of Astragalus and Angelica retarded the progression of renal fibrosis and deterioration of renal function with comparable effects of enalapril. These effects were not caused by blocking the intrarenal renin-angiotensin system, but associated with suppression of the overexpression of TGFbeta1 and osteopontin, reduction of infiltrating macrophages, and less activation of renal intrinsic cells [corrected]. PMID- 14738910 TI - Interleukin-18 induces serum amyloid A (SAA) protein production from rheumatoid synovial fibroblasts. AB - Interleukin-18 (IL-18) is a novel proinflammatory cytokine that was recently found in synovial fluids and synovial tissues from patients with rheumatoid arthritis (RA). To investigate the role of IL-18 in rheumatoid synovitis, the levels of IL-18 and serum amyloid A (SAA) were measured in synovial fluids from 24 patients with rheumatoid arthritis (RA) and 13 patients with osteoarthritis (OA). The levels of IL-18 and SAA in the synovial fluids were elevated in RA patients. In contrast, the levels of IL-18 in synovial fluids from OA patients were significantly lower compared to those of RA patients. SAA was not detected in synovial fluids from OA patients. The expression of SAA mRNA in rheumatoid synovial cells was also examined. SAA4 mRNA, which was constitutively expressed by rheumatoid synovial cells, was not affected by IL-18 stimulation. Although acute phase SAA (A-SAA, SAA1 + 2) mRNA was not detected in unstimulated synovial cells, its expression was induced by IL-18 stimulation. By immunoblot, we demonstrated that IL-18 induced the SAA protein synthesis from rheumatoid synovial cells in a dose-dependent manner. These results indicate a novel role for IL-18 in rheumatoid inflammation through the synovial SAA production. PMID- 14738911 TI - Effect of nitric oxide on prolactin secretion and hypothalamic biogenic amine contents. AB - Involvement of nitric oxide (NO) in the episodic secretion of prolactin was studied in conscious freely moving adult rats. Prolactin secretion was pulsatile in all animals of either group during the bleeding period (from 10:30 h to 13:30 h). Administration of N(omega)-nitro-L-arginine methyl ester (L-NAME), a NO synthase inhibitor, increased mean plasma levels of prolactin, and the absolute amplitude of prolactin peaks during the whole bleeding period as compared to values found in the control group. L-NAME increased norepinephrine (170%), dopamine (58.27%) and serotonin contents (30%) in the anterior hypothalamus. In the median eminence, dopamine and serotonin contents decreased (19.79% and 33.9% respectively) after L-NAME as compared to the values found in controls. In addition, norepinephrine content increased in mediobasal hypothalamus (79.6%) of rats treated with L-NAME. The results indicate that changes in NO production may modify the episodic secretion of prolactin. These effects were associated with changes in hypothalamic and median eminence biogenic amines. PMID- 14738912 TI - Inhibition of xanthine oxidase by phytic acid and its antioxidative action. AB - We examined if phytic acid inhibits the enzymatic superoxide source xanthine oxidase (XO). Half inhibition of XO by phytic acid (IC50) was about 30 mM in the formation of uric acid from xanthine, but generation of the superoxide was greatly affected by phytic acid; the IC50 was about 6 mM, indicating that the superoxide generating domain of XO is more sensitive to phytic acid. The XO activity in intestinal homogenate was also inhibited by phytic acid. However, it was not observed with intestinal homogenate that superoxide generation was more sensitive to phytic acid compared with the formation of uric acid as observed with XO from butter milk. XO-induced superoxide-dependent lipid peroxidation was inhibited by phytic acid, but not by myo-inositol. Reduction of ADP-Fe3+ caused by XO was inhibited by superoxide dismutase, but not phytic acid. The results suggest that phytic acid interferes with the formation of ADP-iron-oxygen complexes that initiate lipid peroxidation. Both phytic acid and myo-inositol inhibited XO-induced superoxide-dependent DNA damage. Mannitol inhibited the DNA strand break. Myo-inositol may act as a hydroxyl radical scavenger. The antioxidative action of phytic acid may be due to not only inhibiting XO, but also preventing formation of ADP-iron-oxygen complexes. PMID- 14738913 TI - Integrated high capacity solid phase extraction-MS/MS system for pharmaceutical profiling in drug discovery. AB - A method is described for use in analysis of samples from pharmaceutical profiling of early drug discovery compounds. The method consists of a high capacity autosampler which injects samples into one of two solid phase extraction columns operated in parallel for alternating trapping, washing and elution into a tandem quadrupole mass spectrometer operated in multiple reaction monitoring (MRM) MS/MS mode. A primary method, which is useful for 80-90% of compounds, and a secondary method, which is useful for a majority of the remaining compounds, are described. No analytical HPLC column is used and the analysis rate is approximately 50 samples/h. Specificity is obtained using MRM analysis. Application of the method for high capacity analysis of metabolic stability samples is described. PMID- 14738914 TI - HPLC and LC-MS studies on stress degradation behaviour of tinidazole and development of a validated specific stability-indicating HPLC assay method. AB - The objective of the current investigation was to study the degradation behaviour of tinidazole under different ICH recommended stress conditions by HPLC and LC MS, and to establish a validated stability-indicating HPLC method. The drug was subjected to stress conditions of hydrolysis, oxidation, photolysis and thermal decomposition. Extensive degradation was found to occur in alkaline medium, under oxidative stress and in the photolytic conditions. Mild degradation was observed in acidic and neutral conditions. The drug was stable to thermal stress. Successful separation of drug from degradation products formed under stress conditions was achieved on a C-18 column using water-acetonitrile (88:12) as the mobile phase. The flow rate was 0.8 ml x min(-1) and the detection wavelength was 310 nm. The method was validated with respect to linearity, precision, accuracy, specificity and robustness. The utility of the procedure was verified by its application to marketed formulations that were subjected to accelerated stability studies. The method well separated the drug and degradation products even in actual samples. The products formed in marketed liquid infusions were similar to those formed during stress studies. PMID- 14738915 TI - Validated specific HPLC methods for determination of prazosin, terazosin and doxazosin in the presence of degradation products formed under ICH-recommended stress conditions. AB - The present paper describes development of stability-indicating high-performance liquid chromatographic (HPLC) assay methods for three alpha-adrenergic-blocker drug substances, namely, prazosin, terazosin and doxazosin, in the presence of degradation products generated from forced decomposition studies. Resolution of drugs from degradation products was obtained using a reversed-phase C-18 column using water/acetonitrile/methanol/glacial acetic acid/diethylamine (25:35:40:1:0.017) as mobile phase for prazosin and terazosin and acetonitrile/water/glacial acetic acid/diethylamine (65:35:1:0.02) for doxazosin. The detection was done at 254 nm. The methods were validated with respect to linearity, precision, accuracy, specificity and robustness. PMID- 14738916 TI - N-way PLS applied to simultaneous spectrophotometric determination of acetylsalicylic acid, paracetamol and caffeine. AB - In this work, a simple and rapid analytical procedure was proposed for simultaneous determination of acetylsalicylic acid (ASA), paracetamol (PRC, also known as acetaminophen) and caffeine (CAF) in pharmaceutical formulations based on multivariate calibration and UV spectrophotometric measurements (210-300 nm). The calibration set was constructed with nine solutions in the concentration ranges from 10.0 to 15.0 microg x ml(-1) for ASA and PRC and from 2.0 to 6.0 microg x ml(-1) for CAF, according to an experimental design. The procedure was repeated at four different pH values: 2.0, 3.0, 4.0 and 5.0. Partial least squares (PLS) models were built at each pH and used to determinate a set of synthetic mixtures. The best model was obtained at pH 5.0. An N-way PLS model was applied to a three-way array constructed using all the pH data sets and enabled better results. This calibration model provided root mean squares errors of prediction (RMSEP) from 11.5 to 35% lower than those obtained with PLS at pH 5.0, depending on the analyte. The results achieved for the determination of these drugs in commercial tablets were in agreement to the values specified by the manufactures and the recovery was between 94.7 and 104.5%. PMID- 14738917 TI - Determination of lisinopril in dosage forms and spiked human plasma through derivatization with 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole (NBD-Cl) followed by spectrophotometry or HPLC with fluorimetric detection. AB - Two sensitive, simple and specific methods based on spectrophotometry and reversed-phase HPLC with fluorimetric detection are described for the determination of lisinopril in dosage forms as well as in spiked human plasma using solid phase extraction (SPE) procedures. Both methods are based on the derivatization of lisinopril with 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole (NBD Cl) in borate buffer of pH 9 to yield a yellow, fluorescent product. The spectrophotometric method depends on measuring the formed yellow color at 470 nm after optimization of the reaction conditions. The HPLC method is based on measurement of the derivatized product using fluorescence detection at 540 nm (excitation at 470 nm). The separation of the derivatized drug, the excess reagent and the internal standard (bumetanide) was performed on a reversed-phase ODS column using isocratic elution with methanol-0.02 M sodium dihydrogen phosphate, pH 3.0 (55:45, v/v) at a flow rate of 1.0 ml/min. The calibration graphs were linear over the concentration ranges 2-20 or 0.02-3.2 microg/ml of lisinopril with minimum detectability of 0.3 and 0.008 microg/ml (6.1 x 10(-7) and 1.7 x 10(-8)M) for the spectrophotometric and the HPLC methods, respectively. The proposed methods were applied without any interference from the tablet excipients for the determination of lisinopril in dosage forms, either alone or co-formulated with hydrochlorothiazide. Furthermore, the use of the HPLC method was extended to the in vitro determination of the drug in spiked human plasma. Interference from endogenous amino acids has been overcomed by using the solid phase extraction technique, the percentage recovery (n=6) was 101.6+/-3.35. PMID- 14738918 TI - Validation of a chiral HPLC assay for (R)-salbutamol sulfate. AB - A fast, reliable and specific method for the screening, confirmation, determination and quantitation of salbutamol enantiomers was developed and validated. The described procedure includes a single robust chiral HPLC determination employing a Teicoplanin stationary phase. The method was evaluated for specificity, robustness, linearity, precision and accuracy. Under the chromatographic conditions of the method, known impurities were separated from the active principle. PMID- 14738919 TI - API-ionspray MS and MS/MS study on the structural characterization of bisbenzylisoquinoline alkaloids. AB - API-ionspray MS and MS/MS techniques have been utilized to elucidate the structures of 20 bisbenzylisoquinoline alkaloids, consisting of 17 diether and three monoether links of two benzyltetrahydroisoquinoline units, which were isolated and identified previously from a variety of Thalictrum sp. (Ranunculaceae family). Apparent protonated molecular ions ([M+H](+)) and very intense doubly-protonated molecular ion ([M+2H](++), 100% of relative abundance) in Q1 Scan MS spectra and prominent as well as diagnostic product ions for the structural information in MS/MS spectra were observed in nanogram quantities for all investigated alkaloids. PMID- 14738920 TI - Validated HPLC determination of 2-[(dimethylamino)methyl]cyclohexanone, an impurity in tramadol, using a precolumn derivatisation reaction with 2,4 dinitrophenylhydrazine. AB - A new method for the determination of 2-[(dimethylamino)methyl]cyclohexanone (DAMC) in Tramadol (as active substance or active ingredient in pharmaceutical formulations) is described. The method is based on the derivatisation of 2 [(dimethylamino)methyl]cyclohexanone with 2,4-dinitrophenylhydrazine (2,4-DNPH) in acidic conditions followed by a reversed-phase liquid chromatographic separation with UV detection. The method is simple, selective, quantitative and allows the determination of 2-[(dimethylamino)methyl]cyclohexanone at the low ppm level. The proposed method was validated with respect to selectivity, precision, linearity, accuracy and robustness. PMID- 14738921 TI - Voltammetric behavior and quantification of the sedative-hypnotic drug chlordiazepoxide in bulk form, pharmaceutical formulation and human serum at a mercury electrode. AB - Chlordiazepoxide is a sedative-hypnotic drug widely employed as a transquilizer and anti-depressant. Its electrochemical behavior in Britton-Robinson (B-R) buffers of pH 2-11 at a mercury electrode has been investigated using dc polarography, cyclic voltammetry and controlled-potential coulometry. Polarograms of the drug in B-R buffers of pH 2-10 exhibited three 2-electron waves, while at pH>10, only a single 4-electron wave was observed. The first, second, and third waves in buffers of pH10) may be due to the reduction of both the N-oxide and C=N centers in a one step. The shift of the E(1/2,) values to more negative potentials upon the increase of pH indicated the involvement of protons in the electrode reaction and that the proton-transfer reaction precedes the electrode process proper. The estimated data indicated that, one proton and two electrons are participated in the rate-determining step of each of the reduced centers. The general sequence of chlordiazepoxide reduction processes via each of its reactant centers may be expressed as: H(+), e, e, H(+)((fast)).Based on the interfacial adsorptive character of the drug onto the mercury electrode, a validated direct square-wave adsorptive cathodic stripping (SWAdCS) voltammetric procedure has been described for the trace determination of the drug in bulk form, tablets and human serum. The procedure did not require sample pretreatment or time-consuming extraction or evaporation steps prior to the assay of the drug. The optimized operational conditions of the proposed procedure have been found to be: accumulation potential E(acc.)=-0.9 V, accumulation time t(acc.)=30s, pulse-amplitude=50 mV, scan increment=10 mV and frequency=120 Hz. The proposed procedure is much more simple, fast, sensitive, costly low and achieved much more lower limits of detection (LOD) (4.4 x 10(-10)M and 6.6 x 10(-10)M) and limits of quantitation (LOQ) (1.5 x 10(-9)M and 2.2 x 10( 9)M), respectively in pharmaceutical formulation and spiked human serum, compared to previously reported methods. PMID- 14738922 TI - Determination of ketotifen in human plasma by LC-MS. AB - Analytical validation of a new liquid chromatographic-mass spectrometric (LC-MS) method for determination of total amount of ketotifen (unchanged and conjugated) in human plasma is presented. Pizotifen was used as an internal standard. An enzyme hydrolysis of conjugated ketotifen was conducted with a combination of beta-glucuronidase and aryl sulfatase. After enzyme hydrolysis a liquid-liquid extraction was performed as a cleaning step. The quantitative determination was obtained using selected ion monitoring (SIM) LC-MS. Chromatographic condition was a combination of reverse phase gradient system and a switching column technique. A satisfactory hydrolysis, acceptable accuracy, improved precision in the linear range from 0.5 to 20.0 ng/ml plasma, absolute recovery of 98.04% for ketotifen and 95.13% for pizotifen and stability for 7 months at -20 degrees C have been achieved. PMID- 14738923 TI - Electrochemical determination of oxytetracycline in veterinary drugs. AB - Conductometric, potentiometric and cyclic voltammetric (CV) titration methods are proposed for determination of oxytetracycline (OTC), commonly used in veterinary. The electrochemical titration of OTC hydrochloride with NH(4)Mo(7)O(24), NaVO(3), NaOH, AgNO(3) and FeCl(3) as titrants are reported. The proposed methods were found to be highly precise, having a relative standard deviation (R.S.D.) below 1.0%. Proposed electrochemical titrations were successfully applied to the assay of commercial preparations: Tetrox, Tetramutin OT and Neox, containing the above mentioned antibiotics. The validity of the methods was tested by the recovery studies of standard addition to pharmaceuticals and results were found to be satisfactory. PMID- 14738924 TI - Microscopic determination of tetracycline based on aluminum-sensitized fluorescence of a self-ordered ring formed by a sessile droplet on glass slide support. AB - A fluorescent microscopic determination of trace amount of tetracycline is reported based on the aluminum-sensitized fluorescence effect of a self-ordered ring formed by a sessile droplet on glass slide support. Since the evaporative loss of the solvent from the edge wedge of the droplet that is spotted on a hydrophobic-treated glass slide, an outward capillary flow of the interior solvent of the droplet occurs. The resulted outward capillary flow then carries the solute to the perimeter of the droplet spot where the solute accumulates to form a fluorescent self-ordered ring (SOR). Depending on the spotted volume of the aluminum-tetracycline chelate solution, different size of SOR with the outer diameter (o.d.) less than 1.1mm and the ring belt width less than 21.6 microm can be obtained. Data analysis for the imaged SOR by using a digitalized CCD camera showed that the chelate molecule across the fluorescent SOR belt section follows a Gaussian distribution, and the maximum fluorescent intensity (I(max)) was found to be proportional to tetracycline content. When a 0.1 microl droplet was spotted on the solid surface, tetracycline in the range of 7.5-800.0 fmol (or 7.9 x 10( 8) to 800.0 x 10(-8)moll (-1)) can be detected, and the limit of detection can reach 0.8 fmol (or 7.9 x 10(-9) moll (-1)). With present method, the contents of tetracycline in capsule, tablet, urine and fresh milk were satisfactorily detected with the recoveries of 97.0-106.5% and RSD of 1.2-4.2%, correspondingly. PMID- 14738925 TI - Sensitive fluorimetric method based on sequential injection analysis technique used for dissolution studies and quality control of prazosin hydrochloride in tablets. AB - This report introduces a fully automated flow system for drug-dissolution studies based on the coupling of the sequential injection analysis (SIA) technique with a conventional dissolution apparatus. The methodology described was used for monitoring of dissolution profiles of prazosin hydrochloride (PRH) in pharmaceutical formulation. The very sensitive fluorimetric detection of PRH was performed at lambda(ex)=244 nm (lambda(em)>or=389 nm). Under the optimal conditions, the calibration curve was linear over the range 0.02-2.43 mg x l(-1) of PRH with R.S.D. 1.89, 1.23, and 1.80% (n=10) when determining 0.02, 1.22, and 2.43 mg x l(-1) of PRH in standard solutions, respectively. Equation of the calibration curve was calculated giving the following values: F=4.108 c-3.9 (n=6), r=0.9996. Detection limit was calculated 0.007 mg x l(-1) of PRH. The dissolution test of Deprazolin tablets was programmed for 60 min, with a continuous sampling rate of 70 h(-1) under conditions required by USP 26. Results obtained by SIA technique compared well with HPLC standard method. PMID- 14738926 TI - A validated, sensitive HPLC method for the determination of trace impurities in acetaminophen drug substance. AB - A high-performance liquid chromatography (HPLC) method has been developed and validated for the simultaneous determination of n-propionyl-p-aminophenol, 3 chloro-4-hydroxyacetanilide, 4'-hydroxyacetophenone, 4-hydroxyacetophenone oxime, 4-acetoxyacetanilide and 4'-chloroacetanilide, the main impurities in acetaminophen drug substance. The chromatographic separation was achieved on an Eclipse XDB-18 reversed-phase column using a gradient elution, being solvent A: 0.01 M phosphate buffer at pH 3.0 and solvent B: methanol. The limit of quantitation (S/N=10:1) was 0.1 microg/ml for each impurity. The coefficients of variation were less than 4% for intra-day and inter-day analyses. The individual recovery of acetaminophen spiked samples ranged from 94 to 104% and the mean recovery for each level from 99 to 103% in the 1-150 microg/ml range for all impurities. The proposed method was successfully applied to the analyses of different lots and different manufactures of acetaminophen drug substance. The proposed method can be used for the routine quality control of acetaminophen. PMID- 14738927 TI - Development and validation of a sensitive, specific, and rapid hybridization ELISA assay for determination of concentrations of a ribozyme in biological matrices. AB - Ribozymes are RNA or modified RNA polymers capable of catalyzing cleavage reactions in target strands RNA, and are under development as human therapeutics. Previous methods used for quantitation of nucleic acid polymers in serum or plasma required extraction of the polymer followed by capillary electrophoresis, HPLC, or gel electrophoresis. These methods are time consuming and lack sensitivity. A bioanalytical method has been developed that does not require extraction of the ribozyme analyte from serum. This technique relies on hybridization of the ribozyme molecule to two complementary biotin and digoxigenin labeled oligonucleotide probes. Serum containing the ribozyme is mixed with the labeled probes, and the mixture is heated at 75 degrees C for 5 min to disrupt the ribozyme secondary structure. Samples are then cooled to permit probe annealing and are added to a streptavidin-coated 96-well plate. The bound complex is detected with an anti-digoxigenin alkaline phosphatase (AP) conjugate using PNPP (p-nitrophenyl phosphate) as a substrate. The amount of colored product is measured on a microtiter plate reader at a wavelength of 405 nm. Concentrations of unknown ribozyme samples are estimated based on a standard curve (0.37-270 ng/ml) prepared in serum. The validated lower and upper limits of quantification are 5.0 and 120 ng/ml, respectively. The assay can be completed in approximately 5h and does not require extraction procedures or electrophoretic/chromatographic separation. It is therefore a simple, sensitive and rapid technique. This assay has been validated and has been used for quantitation of serum levels of the HEPTAZYME ribozyme in mouse, monkey, and human pharmacokinetic studies. PMID- 14738928 TI - Determination of loratadine in human plasma by liquid chromatography electrospray ionization ion-trap tandem mass spectrometry. AB - A sensitive and specific liquid chromatography electrospray ionization ion-trap mass spectrometry (LC-ESI-IT-MS/MS) method has been developed and validated for the identification and quantitation of loratadine in human plasma. After the addition of the internal standard (IS), plasma samples were extracted using isooctane:isoamyl alcohol mixture. The compounds were separated on a prepacked Zorbax phenyl column using a mixture of acetonitrile, 0.20% formic acid as mobile phase. A Finnigan LCQ(DUO) ion-trap mass spectrometer connected to a Waters Alliance high performance liquid chromatography (HPLC) was used to develop and validate the method. The results were within the accepted criteria as stated in the FDA bioanalytical method validation guidance. The method was proved to be sensitive and specific by testing six different plasma batches. Linearity was established for the range of concentrations 0.10-10.0 ng/ml with a coefficient of determination (r(2)) of 0.9998. Accuracy for loratadine ranged from 105.00 to 109.50% at low, mid and high levels. The intra-day precision was better than 10.86%. The lower limit of quantitation (LLOQ) was identifiable and reproducible at 0.10 ng/ml with a precision of 9.84%. The proposed method enables the unambiguous identification and quantitation of loratadine for pharmacokinetic, bioavailability or bioequivalence studies. PMID- 14738929 TI - Method development for determining the iohexol in human serum by micellar electrokinetic capillary chromatography. AB - Iohexol is widely used in clinical laboratories as a non-ionic radiographic contrast medium. Determination of its concentration in blood has a vital meaning in preventing its side effects caused by its retention in the system. A method for determining iohexol in serum by micellar electrokinetic capillary chromatography (MECC) requiring no pretreatment is developed. Electrophoresis is performed for serum samples at 25 kV with a borate buffer (50mM; pH 9.5) containing sodium dodecyl sulfate (50mM) and detection is carried out at 245 nm. Migration time of iohexol is 7.4 min. Linearity (0-1000 mg/l) is good and detection limit is 0.5mg/l (S/N=3). CV of intra-assay precision at a measurement concentration range of 6.2-200.1mg/l is 1.38-4.68% and recovery rate is 96-102%. CV of inter-assay precision is 2.06-5.94% at a measurement concentration range of 10.3-155.4 mg/l. This method is characterized by determination through direct injection of serum samples of super micro-quantity into the capillary, which simplifies the determination procedure in a significant manner and improves the precision and accuracy of determination. PMID- 14738930 TI - LC-MS/MS determination of naringin, hesperidin and neohesperidin in rat serum after orally administrating the decoction of Bulpleurum falcatum L. and Fractus aurantii. AB - To identify and quantify biologically active components in rat serum after orally administrating the decoction of Bulpleurum falcatum L. and Fractus aurantii, one of traditional Chinese medicines (TCMs), high-performance liquid chromatography (HPLC)-tandem mass spectrometry method was developed and validated. The HPLC separation was carried out on a Waters Nova Pak C(18) column using acetonitrile and water as mobile phase after the sample of rat serum was cleaned up with solid phase extraction. Atmospheric pressure chemical ionization in the negative ion mode and selected reaction monitoring (SRM) method was developed to determine the active components. Three flavonoids of hesperidin, neohesperidin and naringin were identified in the serum by comparing their retention times and three independent SRM precursor/product ion transitions with those of corresponding reference standards. The concentrations of naringin, hesperidin and neohesperidin in rat serum determined by SRM measurement were 16.3, 11.9 and 14.3 ng/ml, respectively, after orally administrating the decoction of B. falcatum L. and F. aurantii. This method was validated in terms of recovery, linearity, accuracy and precision (intra- and interday variation). The recoveries from spiked control samples were 93.0, 89.3 and 91.2% for hesperidin, neohesperidin and naringin, respectively. Linearity in rat serum was observed over the range of 2.0-50.0 ng/ml. Percent bias (accuracy) and precision were well within the acceptable range and the relative standard deviation (R.S.D.) of the measured rat serum samples was less than 10% (n=5). PMID- 14738931 TI - Determination of celecoxib in human plasma using solid-phase extraction and high performance liquid chromatography. AB - A simple reversed phase high-performance liquid chromatography (HPLC) method was developed for determination of celecoxib levels in human plasma. The procedure involves solid-phase extraction of celecoxib and the internal standard (SC-236) from plasma using C(18) extraction cartridges. The chromatographic separation of celecoxib and SC-236 was achieved with a Nova Pak C(8) column (3.8 mm x 150 mm) eluted with a mobile phase consisting of acetonitrile-tetrahydrofuran-sodium acetate buffer (pH 5.0) in the ratio of 30:8:62. An ultraviolet light detector with the wavelength set at 215 nm was employed for detection. Celecoxib was well resolved from the plasma constituents and the internal standard. The extraction recovery of celecoxib and SC-236 from human plasma was greater than 88%. Linear calibration curves were established over a concentration range of 40-4000 ng/ml when 0.25 ml aliquots of plasma were used. The inter- and intra-day R.S.D. for the assay was less than 12 and 5%, respectively. This assay has been applied to the analysis of celecoxib levels in plasma samples collected from healthy participants entered into a Phase II clinical study. PMID- 14738932 TI - Quantitative analysis of simvastatin and its beta-hydroxy acid in human plasma using automated liquid-liquid extraction based on 96-well plate format and liquid chromatography-tandem mass spectrometry. AB - An assay based on automated liquid-liquid extraction (LLE) and liquid chromatography-tandem mass spectrometry (LC/MS/MS) has been developed and validated for the quantitative analysis of simvastatin (SV) and its beta-hydroxy acid (SVA) in human plasma. A Packard MultiProbe II workstation was used to convert human plasma samples collected following administration of simvastatin and quality control (QC) samples from individual tubes into 96-well plate format. The workstation was also used to prepare calibration standards and spike internal standards. A Tomtec Quadra 96-channel liquid handling workstation was used to perform LLE based on 96-well plates including adding solvents, separating organic from aqueous layer and reconstitution. SV and SVA were separated through a Kromasil C18 column (50 mm x 2 mm i.d., 5 microm) and detected by tandem mass spectrometry with a TurboIonspray interface. Stable isotope-labeled SV and SVA, 13CD(3)-SV and 13 CD(3)-SVA, were used as the internal standards for SV and SVA, respectively. The automated procedures reduced the overall analytical time (96 samples) to 1/3 of that of manual LLE. Most importantly, an analyst spent only a fraction of time on the 96-well LLE. A limit of quantitation of 50 pg/ml was achieved for both SV and SVA. The interconversion between SV and SVA during the 96-well LLE was found to be negligible. The assay showed very good reproducibility, with intra- and inter-assay precision (%R.S.D.) of less than 7.5%, and accuracy of 98.7-102.3% of nominal values for both analytes. By using this method, sample throughput should be enhanced at least three-fold compared to that of the manual procedure. PMID- 14738933 TI - Fluorescence spectroscopy studies on 5-aminosalicylic acid and zinc 5 aminosalylicylate interaction with human serum albumin. AB - The interaction between 5-aminosalicylic acid (5-ASA) or zinc 5-aminosalylicylate (5-ASZ) and human serum albumin (HSA) was studied by fluorescence spectroscopic technique. The binding constants of 5-ASA or 5-ASZ with HSA were determined at different temperatures under the optimum conditions. The binding sites were obtained and the acting force were suggested to be mainly hydrophobic. The effect of common ions on the binding constants was also investigated. A fluorescence spectroscopy assay of the proteins is presented in the paper. The determination results of the proteins in human serum by this method are very close to those obtained using Coomassie Brilliant Blue G-250 colorimetry, with relative standard deviations of 0.8-2.9%. A practical method was proposed for the determination of 5-ASA or 5-ASZ in human serum samples. PMID- 14738934 TI - HPLC electrospray mass spectrometric characterization of trimeric building blocks for oligonucleotide synthesis. AB - Trimeric nucleotide building blocks are valuable in synthesis of randomized oligonucleotides. In this study, we have developed HPLC-MS and HPLC-MS/MS methods for quality control of protected trinucleotides. C18 reversed-phase HPLC was used for purity evaluation, and base sequences were verified using negative ion electrospray ionization mass spectrometry (ESI-MS) and collision induced dissociation (CID) MS/MS. The principal dissociation pathway was formation of w ions, which represent the 3'-5' direction. The other major fragments were d-ions, which are formed by cleavage of 5' C-O bonds of the sugars. The developed method was suitable for verification of purity and structure of the 29 trinucleotides studied. PMID- 14738935 TI - Determination of combined p-hydroxy benzoic acid preservatives in a liquid pharmaceutical formulation by HPLC. AB - This paper describes a reversed-phase high performance liquid chromatographic (RP HPLC) assay method for the determination of combined p-hydroxy benzoic acid (ethylparaben (EP), methylparaben (MP) and propylparaben (PP)) preservatives in a liquid pharmaceutical formulation. The chromatographic separation was achieved with potassium phosphate buffer (pH 7.05)-methanol (47.5:52.5, v/v) as mobile phase, a Spherisorb C(18) column (250 mm x 4.6mm) and UV detection at 254 nm. The analysis time was <8 min. The method was validated with respect to linearity, precision, accuracy, selectivity, specificity and ruggedness. The calibration curves showed good linearity over the concentration range of 2-140 microg/ml. The correlation coefficient were >0.9999 in each case. The relative standard deviation (R.S.D.) values for intra- and inter-day precision studies were <1%. The procedure describe here is simple, selective and is suitable for routine quality control analysis and stability tests. PMID- 14738936 TI - The effect of eluent pH and compound acid-base character on the design of generic gradient reversed-phase high-performance liquid chromatography (RP-HPLC) methods for use in drug discovery. AB - The design of generic reversed-phase high-performance liquid chromatography (RP HPLC) gradient methods for the analysis of compound mixtures or 'cocktails' has been investigated with particular reference to the eluent pH and the type of compound (acid, base or neutral) analysed. The use of eluents with an acidic eluent pH, an approach which is widely employed, can lead to non-retention of polar bases resulting in 'failure' of the method. This problem is aggravated where the majority of compounds submitted for analysis are bases, which is typical of many drug discovery programs. The problem can be ameliorated through the use of eluents with near neutral pH. Although these neutral pH eluents can lead to co-elution when cocktails are analysed and possibly ion-suppression where mass spectrometry (MS) is the detection method, this can be avoided through optimisation of the gradient shape. PMID- 14738937 TI - Spectrofluorimetric determination of 2-aminopyridine as a potential impurity in piroxicam and tenoxicam within the pharmacopoeial limit. AB - The British Pharmacopoeia defines 2-aminopyridine (2-AP) as a potential impurity in piroxicam (PX) and tenoxicam (TX). Selective spectrofluorimetric determination of 2-AP in PX and TX, within or near the pharmacopoeial level, 0.2%, was developed, based on the measurement of the native fluorescence either in aqueous 0.1N sulfuric acid or in dioxane. Accordingly, this approach was followed for confirming purity of PX and TX in bulk and pharmaceutical preparations. The study was also extended to include simultaneous determinations of PX/2-AP and TX/2-AP systems based on selective fluorescence measurements in the cited solvents. PMID- 14738938 TI - Determination of butein in rat serum by high performance liquid chromatography. AB - A simple high performance liquid chromatography (HPLC) method was developed for the determination of butein in rat serum. The method involved the deproteinization followed by injection into a Luna C8 column. Butein was eluted at 3.8 min at a flow rate of 0.2 ml/min with the mobile phase of acetonitrile ammonium formate (10mM, pH 3.0) (35:65, v/v). The standard curve was linear (r(2)=0.995) over the concentration range of 0.1-10 microg/ml. The coefficient of variation (CV) of intra- and inter-assay ranged from 2.7 to 7.5% and 6.0 to 7.5%, respectively. The limit of quantification was 0.1 microg/ml using a serum sample of 50 microl. This method was applied to a pharmacokinetic study after intravenous injection of butein (5mg/kg) to rats. PMID- 14738939 TI - Quantitative determination of alginic acid in pharmaceutical formulations using capillary electrophoresis. AB - A capillary electrophoresis (CE) method has been developed and validated for the quantitative determination of alginic acid, which is used as a rafting agent in complex antacid formulations. The method involves a preliminary separation of the alginic acid from the formulation by washing the sample matrix with methanol, diluted HCl and water. This is followed by electrophoresis within a fused silica capillary using borate/boric acid buffer as the electrolyte, and the quantification is performed by a UV detector monitoring at 200 nm, where the intrinsic absorption of alginic acid is measured. An assay precision of better than 3% was achieved in intra- and interday determinations. No interference was found from the matrix of the antacid formulations. PMID- 14738940 TI - HPLC determination of sertraline in bulk drug, tablets and capsules using hydroxypropyl-beta-cyclodextrin as mobile phase additive. AB - A sensitive and stereospecific high-performance liquid chromatography (HPLC) method for determination of sertraline in bulk drug, tablets and capsules was developed. Chromatography resolution of the sertraline enantiomeric forms and trans diastereoisomers was performed on Alltima C18 (250 mm x 4.6mm i.d., 5 microm) column with hydroxypropyl-beta-cyclodextrin (HP-beta-CD) as mobile phase additive. The composition of the mobile phase was 68:32 (v/v) aqueous 170 mM phosphate buffer, pH 3.0 (adjusted with 85% phosphoric acid) containing 18 mM HP beta-CD/acetonitrile at a flow rate of 1.0 ml x min(-1). The UV detector was set at 225 nm. Calibration curves were linear (r=0.9999, n=9) in the range of 1-120 microgml (-1) for sertraline. Limit of detection and quantitation for sertraline was 0.029 and 0.097 microg x ml (-1). The values of R.S.D. of repeatability and intermediate precision for bulk drug, tablets and capsules of sertraline hydrochloride were less than 1.0%. PMID- 14738942 TI - Psychogenic movement disorders in children. AB - A common problem in neurology is the existence of disorders that present with neurologic symptoms but do not have an identifiable neurologic basis. These disorders are often thought to have a psychologic basis. Abnormal movements are among the most frequent symptoms in psychogenic neurologic disorders. Although these disorders have not been studied extensively in children, clinical experience in our busy pediatric movement disorders clinic and many case reports support their existence in this age group. Elements of history, physical examination, and therapeutic intervention must be combined to construct a clear diagnosis of a psychogenic movement disorder. This article reviews the diagnosis and treatment of these disorders and includes two illustrative cases. Review of the current literature reveals a need for prospective trials to provide a solid foundation for better diagnosis and treatment of these disorders. PMID- 14738943 TI - Presence of filamin in the astrocytic inclusions of Aicardi syndrome. AB - Aicardi syndrome affects only females and has been hypothesized to be an X-linked dominant male-lethal disorder. Because no familial cases can be studied for genetic linkage analysis, the mutated gene has remained elusive. With the goal of selecting genes for mutation analysis by a functional candidate approach, a detailed pathologic analysis of two brains from deceased Aicardi syndrome patients was performed. The presence of micrencephaly, absent or hypoplastic corpus callosum, polymicrogyria, heterotopia, ventriculomegaly, intracerebral cyst, and intracytoplasmic eosinophilic inclusions was confirmed in glial fibrillary acidic protein-positive astrocytes in the cortex and heterotopias, but not in white matter. The inclusions demonstrated strong immunolabeling with antibodies to filamin and vimentin but weak labeling with antibodies to proteins S100 and microtubule-associated protein 1. These findings suggested that an underlying defect in the cytoskeleton, which involves filamin, may cause this condition. Because the filamin A gene in Xq28 is mutated in another disorder with heterotopia, familial bilateral periventricular heterotopia, mutation analysis of filamin A in Aicardi syndrome patients was pursued. No mutations were found, and the full-length protein was expressed in both brain samples. Future studies will focus on investigation of X-linked genes that may affect function of filamin or other cytoskeletal proteins. PMID- 14738944 TI - Agyria-pachygyria complex: MR findings and correlation with clinical features. AB - The aim of this study was to determine the spectrum of clinical abnormalities in the agyria-pachygyria complex, to identify possible causes, and to correlate the clinical features with the extent of the lesions on magnetic resonance imaging. On the basis of the magnetic resonance imaging findings, 37 patients (22 males, 15 females; mean age 21.1 +/- 31.2 months) with agyria-pachygyria complex were separated into two groups: Group 1 (18 children) manifested generalized or bilateral gyral malformation, and Group 2 (19 children) manifested localized or unilateral gyral malformation. The ratio of generalized seizures in Group 1 was significantly higher, whereas partial seizures were more common in Group 2. Group 1 patients had seizures significantly more frequently than Group 2 patients. Diffuse electroencephalographic abnormalities were significantly more common in Group 1, as were the localized abnormalities in Group 2. Hemipareses were the most frequent neurologic deficit among Group 2 patients. Spastic quadriparesis and microcephaly were more common in Group 1. In conclusion, the extent of agyria pachygyria complex varies widely and the clinical features are accordingly diverse. Patients with bilateral or generalized gyral anomalies have poor prognosis for outcome of epilepsy and neurologic disability. The recognition of these lesions with higher-resolution techniques of magnetic resonance imaging is important for planning proper treatment and genetic counseling. PMID- 14738945 TI - Idiopathic occipital and absence epilepsies appearing in the same children. AB - Our aim is to report the association between idiopathic occipital epilepsy and childhood absence epilepsy in the same children. Six children met the diagnostic criteria for both idiopathic occipital epilepsy and childhood absence epilepsy, five patients with idiopathic occipital epilepsy Gastaut type and another with Panayiotopoulos type. All patients were monitored for 2 to 10 years with repeated electroencephalograms when awake and during sleep. Age at onset of seizures ranged from 4.6 to 8 years. Five patients had focal sensory visual seizures, all with migraine-like episodes. One patient presented ictal vomiting followed by oculocephalic deviation. All patients presented typical absences, with onset at least 1 year after having had idiopathic occipital epilepsy Gastaut type in three patients. In the other two patients with idiopathic occipital epilepsy Gastaut type and the patient with idiopathic occipital epilepsy Panayiotopoulos type, both types of epilepsy appeared at the same time. The electroencephalograms documented occipital paroxysms in all cases, with positive reactivity to the eye closure in five patients. All children presented spike-wave discharges at 3 cycles per second activated by hyperventilation. More genetic information would be necessary to demonstrate either a close genetic relationship between these syndromes or common markers with variable phenotypes. PMID- 14738946 TI - Physician preference for antiepileptic drug concentration testing. AB - A four-item questionnaire asked active U.S. members of the Child Neurology Society to value painless antiepileptic drug concentration monitoring, whether members had ordered a saliva level (the best established painless method) in the last year, and whether such levels were available. Value was quantified by time per patient that the physician would willingly expend to arrange for the test. Of 945 questionnaires sent, 544 (58%) were returned. When asked the value of a painless method for children, 286/522 (55%) reported willingness to expend 10 to 30 minutes to arrange the test; 498/522 (95%) would use a painless method if available. When asked the value of an immediate sample at home during a seizure or adverse event, a substantial majority, 370/526 (70%), would make an important donation of their own time to arrange for the sample. Only 5% would not use it. Just 2/544 respondents had obtained a painless (saliva) concentration, and merely 33/544 (6%) perceived such tests as being available. We conclude that child neurologists put a high value on painless antiepileptic monitoring. These data suggest that a painless method of measuring antiepileptic drug concentrations- especially if it could be performed at home--would fulfill an unmet need in the care of children with epilepsy. PMID- 14738947 TI - Not enough of a good thing. PMID- 14738948 TI - Strategies to attract medical students to the specialty of child neurology. AB - The decline in the number of medical students choosing to enter the field of child neurology is a concern. We undertook this study to learn more about the qualities of highly regarded medical schools that may play a role in attracting students to the field of child neurology. We surveyed child neurologists at top U.S. medical schools that were most successful and least successful at attracting students to child neurology to determine what factors influenced the number of students entering the field of child neurology. We determined that the medical schools that produced the most child neurologists had stronger neuroscience curricula, stronger academic reputations, and larger Divisions of Child Neurology. Our findings suggest that our attention should be focused on academic centers that have more resources to create an atmosphere that is appealing to prospective applicants. These schools should implement a curriculum in neuroscience and child neurology that specifically exposes students early and maintains their interest in the field of child neurology. PMID- 14738949 TI - Brainstem calcification in Mobius syndrome. AB - Mobius syndrome is characterized by congenital facial diplegia, and may be associated with limb or orofacial malformations. A number of mechanisms have been proposed to explain the pathogenesis, including prenatal ischemia. We identified seven children with Mobius syndrome over the 10-year interval 1992-2001, all of whom manifested incomplete bilateral facial palsy. Associated limb and orofacial anomalies were observed in six cases. Computed tomographic scans were available in six children, and five of them manifested brainstem calcification which was most prominent in the floor of the fourth ventricle. The calcification was detected as early as 7 days of age and did not change with time, suggesting a static condition of prenatal onset. These observations support the hypothesis that the pathology in Mobius syndrome is secondary to prenatal brain ischemia. PMID- 14738950 TI - Outcome of very low birth weight infants with sonographic enlarged occipital horn. AB - The objective of this study is to compare the neurodevelopmental outcome between very low birth weight infants with and without sonographic disproportionate enlargement of occipital horn. We retrospectively reviewed the brain sonography of all very low birth weight infants born at National Taiwan University Hospital between June 1997 and June 1999. Brain sonography was routinely performed at the age of the third, seventh, twenty-first, and later days as clinically indicated. Intracranial hemorrhage, periventricular leukomalacia, congenital hydrocephalus, and Stage III retinopathy of prematurity were excluded from our study because of the association with neurodevelopmental impairment. Patients with disproportional dilatation of occipital horn more than 15 mm in width were included in the study group, and those with less than 15 mm were in the control group. Both groups received developmental evaluation by the Bayley Scales of Infant Development II at corrected age of 6, 12, 18, and 24 months, respectively. Socioeconomic status and detailed medical history were obtained at assessments. Independent-samples t test was used for comparison. A total of 81 very low birth weight infants were included in this study: 49 infants (female 18, male 31) in the study group and 32 infants (female 23, male 9) in the control group. The mean gestation in these two groups was 30 +/- 2 weeks and 31.1 +/- 2.2 weeks (P = 0.156), and the mean birth body weight was 1290 +/- 269 gm and 1282 +/- 219 gm (P = 0.877), respectively. At corrected age of 24 months, there was no significant difference in muscle tone, neuromotor impairment, hearing impairment, vision, or speech development. Assessment with the mental development index (88.9 +/- 15.6 vs 93 +/- 13.2) (P = 0.238) and the psychomotor development index (93.3 +/- 10.3 vs 89.6 +/- 12.1) (P = 0.149) between these two groups was also comparable. This retrospective analysis suggests that ultrasonographic disproportionate enlargement of the occipital horn in very low birth weight infants does not affect the neuromotor development at corrected ages of 6, 12, 18, and 24 months. PMID- 14738951 TI - Subependymal veins in premature neonates: implications for hemorrhage. AB - The germinal matrix contains a concentrated network of blood vessels. The unusual structural qualities of these vessels are implicated as a factor underlying the high incidence of hemorrhage that occurs in the germinal matrix of prematurely born neonates. The present study is a histologic analysis of an postmortem examination series of brains collected from neonates born between 23 weeks gestation and term and is designed to determine if subependymal veins can be recognized in neonates born at the limits of viability, approximately 23 weeks gestation. Alkaline phosphatase histochemistry is used to differentiate cerebral afferent from efferent vessels. The results demonstrate that precursors of the subependymal veins can be recognized as early as the twenty-third gestational week. These veins increase progressively in diameter from 23 weeks to term, but the wall of the veins, which at early stages consists of endothelial cells only, does not thicken until after postconception week 36. Thus in all premature neonates, including the youngest capable of independent existence, the subependymal veins are present and appear vulnerable to rupture. These data support our suggestion that the structural immaturity of these veins in premature neonates is causally related to the high incidence of germinal matrix hemorrhage in these patients. PMID- 14738952 TI - Cervical myelitis from herpes simplex virus type 1. AB - Although subacute ascending paralysis without sensory involvement is typically evocative of Guillain-Barre syndrome, it can alternatively be due to infection or inflammation of the spinal cord. We describe a 16-month-old female who presented with ascending flaccid paresis after an upper respiratory tract infection. She then developed signs of upper motor neuron involvement of the lower limbs associated with upper motor neuron involvement of the upper limbs. Motor nerve conduction and electromyographic studies of upper limbs demonstrated anterior horn cell involvement. Neuroimaging was consistent with cervical myelitis, and cerebrospinal fluid polymerase chain reaction was positive for herpesvirus-1. Although association with the primary infection of the respiratory tract may be fortuitous, possible neurotropic or hematogenous spread of herpesvirus-1 to the cervical spinal cord cannot be excluded. She then developed signs of upper motor neuron involvement of the lower limbs associatred with lower motor neuron involvement of the upper limbs [corrected]. PMID- 14738953 TI - Atypical childhood Wilson's disease. AB - Wilson's disease is a genetic disorder of copper metabolism with a hepatic or neurologic presentation. A hepatic presentation is more common in young children. Neurologic Wilson's disease often manifests as a movement disorder with dystonia, tremor, and dysarthria. Psychiatric or behavioral symptoms can also be a presenting feature of Wilson's disease. We describe an atypical neurologic presentation in a prepubertal child with minimal hepatic involvement; in which transient hemiparesis and encephalopathy dominated her initial neurologic presentation. Brain magnetic resonance imaging revealed extensive cortical and subcortical signal change, in addition to the classical basal ganglia signal abnormality observed in Wilson's disease. She was treated with oral tetrathiomolybdate anticopper therapy, followed by zinc maintenance. Her clinical status and brain imaging improved considerably at 1 year after treatment initiation. Neurologic Wilson's disease may have diverse presentations, and should be considered in children who present with cortical features and signal change on magnetic resonance imaging. PMID- 14738954 TI - Cohen syndrome with insulin resistance and seizure. AB - Cohen syndrome is a rare, genetic, connective-tissue disorder with the genetic abnormality linked to chromosome 8q22. The diagnosis of Cohen syndrome is based on the recognition of certain clinical findings, which include mental retardation, typical morphologic stigmata (e.g., truncal obesity, hypotonia, short philtrum, prominent frontal incisors, high-arched palate, narrow hands and feet), and characteristic ophthalmologic abnormalities. We report a patient manifesting the typical characteristics of Cohen syndrome with seizure and hyperinsulinemia. PMID- 14738955 TI - Ophthalmoplegic migraine. AB - Ophthalmoplegic migraine is a rare presentation of migraine complicated by an isolated oculomotor paresis. Vasodilation of extracranial vessels is believed to underlie the headache, and vasoconstriction to account for the ophthalmoplegia. Whether the vascular insult involves the central or peripheral portions of the oculomotor nerve is still uncertain. We describe a child who presented with ophthalmoplegic migraine and was demonstrated to have a deficiency of the near triad documented by eye movement and pupillary recordings. Voluntary conjugate eye movements--saccades, smooth pursuit, and optokinetic nystagmus--were normal. Vergence amplitudes appropriate to fixation distance were elicited for Snellen optotypes but not to a point source of light. Concurrent measures of pupillary diameter failed to reveal significant modulation to either stimulus. Brain magnetic resonance imaging scan was normal, and there was no contrast enhancement of the oculomotor nerve at its exit from the midbrain. Both the oculomotor paresis and concurrent presence of a deficiency of the near triad localized the vascular insult to the oculomotor nerve complex in the brainstem. PMID- 14738956 TI - Temporo-occipital spikes: a typical EEG finding in Kabuki syndrome. AB - Kabuki syndrome is a rare dysmorphogenic disorder. The central nervous system is often involved, and epilepsy is a common symptom. The diagnosis is clinical, and no typical electroencephalographic findings have thus far been reported. We have documented temporo-occipital spikes in sleep electroencephalogram in all our three Kabuki patients. The location of the spikes was similar in all cases although their occurrence varied from continuous spiking to single spikes. We suggest that temporo-occipital spikes are typical in Kabuki syndrome and discuss the possible cause of this finding. PMID- 14738957 TI - Transient cerebellar mutism in the course of acute cerebellitis. AB - Transient mutism after posterior fossa surgery in children or associated with cerebellar hemorrhage or trauma is a recognized phenomenon. However, its association with parainflammatory cerebellitis has been rarely described. We report on a previously healthy 3-year-old child with severe cerebellitis after acute gastroenteritis of unidentified cause. Severe ataxia and transient mutism were the prevailing clinical features. Magnetic resonance imaging revealed swelling of the cerebellum with protruding cerebellar tonsils at the level of the occipital foramen. Recovery from the acute illness was slow and incomplete. Residual cerebellar dysfunction manifested with dysphonic and dysarthric speech, as well as motor coordination problems and was associated with atrophy of the vermis and cerebellar hemispheres in follow-up studies. PMID- 14738958 TI - Duane's syndrome and giant parietal foramina. AB - We report a 14-month-old male with Duane's syndrome and bilateral parietal calvarial defects. This child had no other known medical conditions. The Duane's syndrome involved the left eye. Imaging was demonstrative of bilateral enlarged parietal foramina. We believe this to be the first reported case of simultaneous Duane's syndrome and bilateral giant parietal foramina. Further case reports of our current association are required to determine if these simultaneous findings are spurious or a low-frequency association. PMID- 14738960 TI - A potential link between measles virus and autism: age-matched control groups are essential. PMID- 14738961 TI - The interaction of glycine, aspartic acid, and lysine by the protonated macrocyclic ligand 6,19-bis(2-hydroxypropyl)-3,6,9,16,19,22-hexaaza-tricyclo [22.2.2.2(11,14)]triaconta-11,13,24,26,27,29-hexaene. AB - A new hexaaza macrocyclic ligand (L) bearing two 2-hydroxypropyl pendants, 6,19 bis(2-hydroxypropyl)-3,6,9,16,19,22-hexaaza-tricyclo-[22.2.2.2(11,14)]triaconta 11,13,24,26,27,29-hexaene has been synthesized and characterized. The macrocyclic ligand was isolated as a colorless crystal, monoclinic, P2(1)/n, with a=10.757(2), b=14.214(3), c=13.746(3) A, beta=101.40(3) degrees, V=2060.3(7) A3, Z=2, R1=0.0695, and wR2=0.1538 [I>2sigma(I)]. Potentiometric studies of the macrocyclic ligand and three types of amino acids, glycine (equal numbers of carboxylate and amino groups), aspartic acid (more carboxylate groups than amino group), and lysine (more amino groups than carboxylate group) have been performed. The stability constants for the new macrocycle and binary complexes of the amino acid with the macrocyclic ligand are reported. Binary complexes are formed in aqueous solution as a result of hydrogen bonding interaction and electrostatic attraction between the host and the guest. The binding Schemes for the recognition of amino acids are suggested. From the results, it seems that this new macrocyclic ligand is able to bind three different amino acids with selectivity in aqueous solution, and the strength of binding is of the order lysine < glycine < aspartic acid. PMID- 14738962 TI - Cytotoxic and antimalarial constituents from the roots of Eurycoma longifolia. AB - Sixty-five compounds were isolated from the roots of Eurycoma longifolia and characterized by comprehensive analyses of their 1D and 2D NMR, and mass spectral data. Among these isolates, four quassinoid diterpenoids were reported from natural sources for the first time, namely eurycomalide A (1), eurycomalide B (2), 13beta, 21-dihydroxyeurycomanol (3), and 5alpha, 14beta, 15beta trihydroxyklaineanone (4). Screening of cytotoxicity, anti-HIV and antimalarial activity of these isolated compounds was also furnished by in vitro assays. Compounds 12, 13, 17, 18, 36, 38, 59, and 62 demonstrated strong cytotoxicity toward human lung cancer (A-549) cell lines, however, 12, 13, 17, 38, 57, 58, and 59 exhibited strong cytoxicity toward human breast cancer (MCF-7) cell lines. Compounds 57 and 58 displayed potent antimalarial activity against the resistant Plasmodium falciparum. The thorough studies on the stereochemistry of the different quassinoid diterpenoids provide a clear reference to the scientists who are interested on this field. PMID- 14738963 TI - General strategy for the preparation of membrane permeable fluorogenic peptide ester conjugates for in vivo studies of ester prodrug stability. AB - To study ester prodrug stability properties in living cells we have conjugated fluorogenic esters to the cell membrane permeable peptide Arg9. The desired conjugates are prepared by coupling N-maleoyl amino acid esters of monoalkylated fluoresceins or fluorescein to TyrArg9Cys. The photophysical properties of the monoalkylated fluorescein derivatives are described. PMID- 14738964 TI - Synthesis and pharmacological evaluation of 5H-[1]benzopyrano[4,3-d]pyrimidines effective as antiplatelet/analgesic agents. AB - Synthesis and pharmacological screening of new 2-methylthio/2-methanesulfonyl/2 methoxy-5H-[1]benzopyrano[4,3-d]pyrimidines were planned in order to study the effects of the 5-substitution with alkoxy/phenoxy/alkylthio and phenylthio groups both on in vitro antiplatelet and in vivo antinociceptive activities. Antiplatelet activity was assessed in vitro against ADP, Arachidonic acid and U46619 induced aggregation, in rabbit plasma. Anti-inflammatory, analgesic and antipyretic activities were tested in rat paw edema, mouse writhing test and LPS induced rat fever, respectively. Amongst test compounds, 2-methylthio derivatives displayed an ASA-like antiplatelet activity whereas 2-methoxy and, particularly, 2-methanesulfonyl derivatives showed a broad spectrum of antiplatelet action, inhibiting both the ADP- and the AA- and U46619-induced aggregation. With regard to the in vivo pharmacological activities, mainly the 2-methoxy derivatives showed a significant analgesic effect comparable to that of indomethacin. SAR considerations, also in comparison with a number of previously described compounds, were performed. PMID- 14738965 TI - Investigations using immunization to attenuate the psychoactive effects of nicotine. AB - Despite the enormous health risks, people continue to smoke and use tobacco primarily as a result of nicotine addiction. As part of our immunopharmacotherapy research, the effects of active and passive immunizations on acute nicotine induced locomotor activity in rats were investigated. To this end, rats were immunized with either a NIC-KLH immunoconjugate vaccine designed to elicit an antinicotine immune response, or were administered an antinicotine monoclonal antibody, NIC9D9, prior to a series of nicotine challenges and testing sessions. Vaccinated rats showed a 45% decrease in locomotor activity compared to a 16% decrease in controls. Passive immunization with NIC9D9 resulted in a 66.9% decrease in locomotor activity versus a 3.4% decrease in controls. Consistent with the behavioral data, much less nicotine was found in the brains of immunized rats. The results support the potential clinical value of immunopharmacotherapy for nicotine addiction in the context of tobacco cessation programs. PMID- 14738966 TI - In vitro study of some medicinally important Mannich bases derived from antitubercular agent. AB - Biologically active Mannich bases with heteroaromatic ring system have been synthesised employing Mannich reaction of isonicotinyl hydrazide with various sulphonamides/secondary amines. They were analysed by elemental analysis and characterized by uv, ir, 1H nmr spectroscopic studies. The Mannich bases were screened for their antibacterial activity against various gram positive and gram negative bacteria and were analyzed statistically. The results have shown that the compounds are quiet active against pathogens under study and were nontoxic. PMID- 14738967 TI - The role of the conserved histidine-aspartate pair in the 'base-off' binding of cobalamins. AB - The conserved cobalamin-binding domain of glutamate mutase exists as a separate dissociable subunit, MutS. The results obtained from BIAcore analysis indicate that MutS alone, in the absence of E component of glutamate mutase (MutE, catalytic subunit), is capable of binding hydroxocobinamide (OHCbi) with a Kd of 15.4+/-1.6 microM, but fails to bind adenosylcobalamin (AdoCbl). The UV-visible spectrum indicates that histidine ligation to the cobalt atom only occurs when both MutE and MutS are present in the solution. MutS mutants, MutS-D14N and MutS H16G, are also capable of binding OHCbi, but their binding kinetics altered. Our experimental results show that the electrostatic interaction between histidine aspartate pair is important in the binding of OHCbi or AdoCbl, no matter whether histidine coordinates to the cobalt atom or not. The catalytic subunit is also involved in histidine ligation to the cobalt atom. Meanwhile, mutation of either His16 or Asp14 significantly impairs the enzyme to cleave the cobalt-carbon bond of AdoCbl. PMID- 14738968 TI - Synergistic effects of anacardic acids and methicillin against methicillin resistant Staphylococcus aureus. AB - The synergistic effects of 6-alk(en)ylsalcylic acids, also known as anacardic acids, in combination with methicillin against Staphylococcus aureus ATCC 33591 (MRSA) was investigated. The double bond in C15-anacardic acids is not essential in eliciting the antibacterial activity but is associated with increasing the activity. The synergistic effects decreased with increasing the number of double bonds in the alkyl chain. On the other hand, the antibacterial activity of anacardic acids possessing different alkyl chain lengths against the same MRSA strain was found to be a parabolic function of their lipophilicity and maximized with the alkyl chain length of C10 and C12. Notably, the synergistic effects were noted to increase with increasing the alkyl chain length. PMID- 14738969 TI - Design, synthesis, and in vitro evaluation of inhibitors of human leukocyte elastase based on a functionalized cyclic sulfamide scaffold. AB - The design of novel functionalized templates capable of binding to the active site of serine proteases could potentially lead to the development of potent and highly selective non-covalent inhibitors of these enzymes. Using the elastase turkey ovomucoid inhibitor complex and insights gained from earlier work based on the 1,2,5-thiadiazolidin-3-one 1,1 dioxide scaffold (I), a surrogate cyclosulfamide scaffold (II) was used for the first time in the design of reversible inhibitors of human leukocyte elastase. Compounds 7 and 8 were found to be micromolar reversible inhibitors of the enzyme. PMID- 14738970 TI - A colormetric assay for catechol-O-methyltransferase. AB - A series of catechol diazo dyes were synthesized and tested as substrates for the enzyme catechol-O-methyltransferase (COMT) with the aim of developing a sensitive HPLC assay method using visible wavelength light detection. A method was developed which allowed for the determination of the two regioisomeric methylated products of the COMT catalyzed reaction of 4-[(3,4 dihydroxyphenyl)azo]benzenesulfonate with S-adenosylmethionine (AdoMet). Separation of the assay components was achieved by reverse phase chromatography using an isocratic mobile phase. No pre-preparation of the assay samples was required. PMID- 14738971 TI - 11C-Radiosynthesis and preliminary human evaluation of the disposition of the ACE inhibitor [11C]zofenoprilat. AB - (4S)-1-[(S)-3-Mercapto-2-methylpropanoyl]-4-phenylthio-L-proline (Zofenoprilat, 2), the active metabolite of the potent ACE inhibitor Zofenopril Calcium (1), was labelled with carbon-11 (t1/2=20.4 min) to evaluate its pharmacokinetics behaviour in human body using Positron Emission Tomography (PET). [11C]2 labelling procedures were based on the use of immobilized Grignard reagent and the acylation of (S)-4-phenylthio-L-proline methyl ester (5) with 11C-labelled methacryloyl chloride, followed by a Michael addition with thiobenzoic acid. The radiochemical yield was 5-10% (EOB, decay corrected) and specific radioactivity ranged from 0.5 to 1.5 Ci/micromol (18.5-55.5 GBq/micromol). Preliminary in vivo human evaluation of [11C]2 showed that the drug accumulates in organs which express high levels of ACE, like lungs and kidneys, and in organs involved in drug metabolism such as the liver and gall bladder. Results of the distribution of [11C]2 showed a measurable concentration of the drug in the target tissues such as the kidney and to a minor extent, the heart, where it can afford organ protection. PMID- 14738972 TI - Structure-activity relationships of thiazole and thiadiazole derivatives as potent and selective human adenosine A3 receptor antagonists. AB - 4-(4-Methoxyphenyl)-2-aminothiazole and 3-(4-methoxyphenyl)-5-aminothiadiazole derivatives have been synthesized and evaluated as selective antagonists for human adenosine A3 receptors. A methoxy group in the 4-position of the phenyl ring and N-acetyl or propionyl substitutions of the aminothiazole and aminothiadiazole templates displayed great increases of binding affinity and selectivity for human adenosine A3 receptors. The most potent A3 antagonist of the present series, N-[3-(4-methoxy-phenyl)-[1,2,4]thiadiazol-5-yl]-acetamide (39) exhibiting a Ki value of 0.79 nM at human adenosine A3 receptors, showed antagonistic property in a functional assay of cAMP biosynthesis involved in one of the signal transduction pathways of adenosine A3 receptors. Molecular modeling study of conformation search and receptor docking experiments to investigate the dramatic differences of binding affinities between two regioisomers of thiadiazole analogues, (39) and (42), suggested possible binding mechanisms in the binding pockets of adenosine receptors. PMID- 14738973 TI - Investigation into new anticonvulsant derivatives of alpha-substituted N benzylamides of gamma-hydroxy- and gamma-acetoxybutyric acid. Part 5: search for new anticonvulsant compounds. AB - A series of four N-benzylamides of gamma-hydroxybutyric acid (GHB), that contain N-(4-phenylpiperazine)-, N-(4-benzylpiperazine)rings, N-benzylamino-, or N-(2 phenylethylamine)-groups in the alpha-position of GHB were selected as model compounds, for determining the structural elements responsible for their potential anticonvulsant action. Based on the results of pharmacological, physicochemical, and molecular modelling investigations, the pharmacophore model for anticonvulsant N-substituted amides of GHB was defined. In this model, the presence of the N-benzylamide fragment is essential for activity. In addition, all of the amides contained another hydrophobic unit (aryl ring) as a distal binding site and H-bond donor. In consideration of these model parameters, a number of N-substituted amides of GHB, containing a hydrophobic moiety such as: N benzylamino or N-(4-chlorobenzylamino) group in the alpha-position of GHB, and a lipophilic substituent in the amide portion, were prepared. It has been shown that the anticonvulsant activities of the newly synthesized compounds might partially be explained on the basis of their lipophilicity (calculated log P values) and the presence of a hydroxyl group in the molecule. PMID- 14738974 TI - Hydropathic interaction analyses of small organic activators binding to antithrombin. AB - Recently we designed the first small organic ligands, sulfated flavanoids and flavonoids, that act as activators of antithrombin for accelerated inhibition of factor Xa, a key proteinase of the coagulation cascade [Gunnarsson and Desai, Bioorg. Med. Chem. Lett. (2003) 13:579]. To better understand the binding properties of these activators at a molecular level, we have utilized computerized hydropathic interaction (HINT) analyses of the sulfated molecules interacting in two plausible electropositive regions, the pentasaccharide- and extended heparin-binding sites, of antithrombin in its native and activated forms. HINT analyses indicate favorable multi-point interactions of the activators in both binding sites of the two forms of antithrombin. Yet, HINT predicts better interaction of most activators, except for (-)-catechin sulfate, with the activated form of antithrombin than with the native form supporting the observation in solution that these molecules function as activators of the inhibitor. Further, whereas (+)-catechin sulfate recognized the activated form of antithrombin better in both the pentasaccharide- and extended heparin- binding sites, the native form was better recognized by (-)-catechin sulfate, thus explaining its weaker binding and activation potential in solution. A reasonable linear correlation between the overall HINT score and the solution free energy of binding of the sulfated activators was evident. This investigation indicates that HINT is a useful tool in understanding interactions of antithrombin with small sulfated organic ligands at a molecular level, has some good predictive properties, and is likely to be useful for rational design purposes. PMID- 14738975 TI - Synthesis of condensed quinolines and quinazolines as DNA ligands. AB - Among new condensed quinolines and quinazolines the design of which were inspired by anti-cancer DNA-binding alkaloids such as camptothecin and batracyclin, DNA binding tests identify the 8-methoxy-7-piperazinylpropoxyindeno[1,2-b]quinolin-11 one tetracyclic system as a new motif for DNA recognition. PMID- 14738976 TI - Development and application of a capsid VP1 (region D) based reverse transcription PCR assay for genotyping of genogroup I and II noroviruses. AB - Noroviruses (NoV), previously called "Norwalk-like viruses", have emerged as the single most important cause of acute gastroenteritis worldwide. Most diagnostic reverse transcription-polymerase chain reaction (RT-PCR) assays target the viral RNA-dependent RNA polymerase; however, the major capsid protein (VP1) is the reference genomic region for establishing genotypes. In this study, we analyzed complete NoV VP1 sequences (n=100) and determined a region (region D) that was most suitable to differentiate between genotypes. Within region D, we designed two genogroup specific, broadly reactive, degenerate primer sets (GI and GII). The region D primers were evaluated in a single-tube one-step RT-PCR assay using a panel of 81 (31 GI, 50 GII) NoV strains from both outbreaks and sporadic cases. In total, 95% of the samples tested positive using the new region D primer sets. Phylogenetic analysis of region D sequences (36 deduced amino acids for GI, 56 deduced amino acids for GII), revealed 19 clusters (7 within GI and 12 within GII) including three new genetically distinct clusters, two of which were unresolved using region A sequences. Phylogenetic analysis of the complete VP1 sequences revealed identical grouping of strains and confirmed the newly identified clusters using region D. In summary, we successfully developed and evaluated a broadly reactive RT-PCR assay for reliable genotyping of GI and GII noroviruses. PMID- 14738977 TI - Adaptation of the plaque assay methodology for dengue virus infected HepG2 cells. AB - The HepG2 cell line is a useful tool for studying dengue virus-cell interactions but as it grows in clumps rather than monolayers, it does not readily adapt itself to the standard plaque assay technique. We therefore sought to develop an indirect plaque assay methodology. Initially HepG2 cells were infected with dengue virus serotype 2 and post-infection incubated for between 0 and 16 h before being treated with trypsin to separate the cells, followed by dilution and plating onto pre-grown monolayers of Vero cells in six well plates. After 7 days incubation and crystal violet staining, plaques were observed at all time points, although there was a relationship between number of plaques and post-infection incubation time, with the longest post-infection incubation time giving the highest number of plaques. To validate the assay with respect to virus input, the experiment was repeated at both the 0 and 16 h post-infection incubation times with different virus: cell levels. At both post-infection incubation times the response of input virus to plaque number was linear. This is a useful adaptation of the plaque assay methodology and one that may be applicable to other virus/cell line combinations. PMID- 14738978 TI - A simple and efficient method for testing Lettuce mosaic virus resistance in in vitro cultivated lettuce. AB - The potential of a new in vitro inoculation and propagation method developed for Lettuce mosaic virus (LMV) on lettuce (Lactuca sativa L.) was evaluated by studying LMV infection on in vitro cultivated seedlings or on newly regenerated plantlets. Lettuce cultivars differing by their LMV-resistance status were inoculated with various natural LMV isolates as well as with Green Fluorescent Protein (GFP)-tagged recombinant virus isolates. A good correlation was observed between the known resistance status of the cultivars and the results obtained by in vitro screening. The results show that this resistance assay can be greatly improved by the use of GFP-tagged virus isolates. The main advantages of this method are reduced space requirements and an improved environmental safety, especially for the handling of recombinant virus, of quarantine virus or of transgenic plants. PMID- 14738979 TI - Development of an in situ hybridisation procedure for the detection of sole aquabirnavirus in infected fish cell cultures. AB - An in situ hybridisation (ISH) technique has been developed to detect sole aquabirnavirus in infected fish cell lines bluegill fibroblast (BF-2), EPC, and chinook salmon embryo cells (CHSE-214). A 613 bp cDNA probe for viral RNA coding for a fragment of VP2 protein was generated by reverse transcription polymerase chain reaction (RT-PCR) using infectious pancreatic necrosis virus (IPNV) specific DNA primers. Infected cells were strongly labelled, and no non-specific reaction was observed in non-infected cells used as negative controls. The specificity of the probe was examined by testing it against a range of IPNV serotypes such as Ab, Sp and VR-299. The ISH technique was compared with the immunofluorescence procedure to determine the sensitivity of detection of sole aquabirnavirus in BF-2 cells. The probe used in the ISH technique detected weak positivity at 8h post-inoculation (p.i.) in the cytoplasm of infected BF-2 cells inoculated with 10(3) TCID50/ml, whilst the labelling appears at 24h p.i. when the immunofluorescence technique was applied. At all other time intervals the results were equivalent. PMID- 14738980 TI - Development of a real-time RT-PCR assay for the detection of potato spindle tuber viroid. AB - Potato spindle tuber viroid (PSTVd) is a quarantine pathogen in the European Union and causes damaging diseases of solanaceous crops. Under the EU Plant Health directive 2000/29/EC, countries must have the ability to detect and identify accurately and rapidly the introduction of harmful organisms in plants or plant products; furthermore, if the quarantine pathogen is found, be able to survey extensively for it. In this respect, PSTVd poses an interesting technical problem, since its RNA does not code for any proteins and thus any diagnostic method must be based on the detection of the RNA and be suitable for scaling up to testing large sample numbers. With this in mind a one-tube real-time RT-PCR assay based on TaqMan chemistry was developed. Investigations were carried out into various aspects of the assay relevant to the efficient amplification of targets that have a significant amount of secondary structure such as viroids. Thus comparisons were made of reverse transcription temperature, concentration and type of reverse transcriptase, RNA denaturation, sample purity and single versus two-tube reaction format. The assay developed was shown to be able to detect a wide range of isolates of PSTVd and in comparison with a chemi luminescent hybridisation system was shown to be 1000-fold more sensitive. A further significant advantage of this assay format compared with hybridisation is that it is suitable for scaling up to large sample numbers using robotic liquid handling systems. PMID- 14738981 TI - Detection and quantitation of akabane and aino viruses by multiplex real-time reverse-transcriptase PCR. AB - A multiplex, quantitive reverse-transcriptase real-time PCR, using MGB TaqMan chemistry, for detecting akabane virus (AKAV) and aino virus (AINV) is described. Each specific probe was labeled with a different fluorescent dye--VIC for detecting AKAV and 6-carboxy-fluorescein (FAM) for detecting AINV. All available sequences of viral S RNA were aligned and primers and probes were designed so that AKAV primers and probes would recognize all AKAVs but not AINV, and vice versa. The parameters for multiplex reactions enabled the detection of both viruses in one tube reaction with similar efficiency. To quantitate the viruses, cDNA amplicons containing the real-time amplicon were prepared using forward primers carrying the T7 promoter sequences. The cDNAs were used directly as templates for run-off transcription and 10-fold dilutions of the products served as standards to quantitate unknown viral samples. Using this system had shown that it could detect approximately 3-30 copies of viral S genome. PMID- 14738982 TI - Comparison of ELISA for the detection of porcine serum antibodies to non structural proteins of foot-and-mouth disease virus. AB - Three foot-and-mouth disease virus non-structural protein antibody detection kits, CHEKIT FMD-3ABC, UBI FMD NS EIA and DVIVR NSP ELISA, were compared in the study. The results showed that the specificity of the kits ranged from 96.7 to 100% in nai;ve pigs and from 93.6 to 98.1% in vaccinated pigs, and that the DVIVR kit had the highest analytical sensitivity. The kappa statistics for the detection of 612 sera were 0.582, 0.447 and 0.658 for CHEKIT/UBI, CHEKIT/DVIVR and UBI/DVIVR, respectively. This study also revealed that measurable non structural protein specific antibodies in some of infected pigs were sustained either for shorter periods or in intermittent patterns, thus aggravating the difficulties associated with the removal of pre-exposed pigs in the field. PMID- 14738983 TI - Expression and purification of turkey coronavirus nucleocapsid protein in Escherichia coli. AB - Purification of turkey coronavirus (TCoV) nucleocapsid (N) protein, expressed in a prokaryotic expression system as histidine-tagged fusion protein is demonstrated in the present study. Turkey coronavirus was partially purified from infected intestine of turkey embryo by sucrose gradient ultracentrifugation and RNA was extracted. The N protein gene was amplified from the extracted RNA by reverse transcription-polymerase chain reaction and cloned. The recombinant expression construct (pTri-N) was identified by polymerase chain reaction and sequencing analysis. Expression of histidine-tagged fusion N protein with a molecular mass of 57 kd was determined by Western blotting analysis. By chromatography on nickel-agarose column, the expressed N protein was purified to near homogeneity as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis. The protein recovery could be 2.5 mg from 100 ml of bacterial culture. The purified N protein was recognized by antibody to TCoV in Western blotting assay. The capability of the recombinant N protein to differentiate positive serum of turkey infected with TCoV from normal turkey serum was evident in enzyme-linked immunosorbent assays (ELISA). These results indicated that the expressed N protein is a superior source of TCoV antigen for development of antibody-capture ELISA for detection of antibodies to TCoV. PMID- 14738984 TI - Evaluation of real-time reverse transcription polymerase chain reaction assays for the detection of swine vesicular disease virus. AB - Differential detection of swine vesicular disease virus (SVDV) from the other vesicular disease viruses of foot-and-mouth disease (FMD), vesicular stomatitis (VS) and vesivirus is important as the vesicular lesions produced by these viruses are indistinguishable in pigs. Two independent sets of primers and probe, designed from nucleotide sequences within the 5' untranslated region (UTR) of the SVDV genome, were evaluated in a real-time (5' nuclease probe-based or fluorogenic) PCR format. Although both primers/probe sets failed to detect one isolate, the assays successfully amplified RNA extracted from epithelial suspensions (ES) and cell culture grown virus preparations from clinical samples representing all currently designated phylogenetic groups of SVDV. Furthermore, no cross-reactivity was demonstrated when these primer/probe sets were tested with RNA prepared from all seven serotypes of FMD virus (FMDV) and from selected isolates of VS virus (VSV), vesivirus and teschoviruses. These assays provide sensitive and rapid alternatives to supplement the routine procedures of ELISA and virus isolation for SVDV diagnosis. The two independent sets of primers/probe can be used routinely while only one of the primers/probe sets would typically be used in SVDV diagnosis during an outbreak. PMID- 14738985 TI - Identification of peptide sequences that induce the transport of phage across the gastrointestinal mucosal barrier. AB - To investigate whether specific peptide sequences could induce virion transport across the intestinal barrier, we used phage display to both identify signalling peptides capable of inducing trans-intestinal transport and also provide a suitable model of virion translocation. We utilised simple, single-round high input in vivo biopanning protocol using a 7-mer random amino acid phage display library. Phage were applied by gavage and translocation across the intestinal barrier assessed by phage recovery from the spleen 2h later. Following isolation, a number of phage were sequenced and several homologies with HIV gp120 were identified. Immunocytochemical analysis of phage translocation across the intestinal barrier by a phage bearing the peptide YPRLLTP demonstrated that phage were actively transported along specific channels. It is concluded that utilisation of in vivo phage display (IVPD) has provided evidence for a specific peptide-guided transport of undegraded cargo across the intestinal barrier, modelled by M13 phage. PMID- 14738986 TI - Use of reverse transcription and PCR to discriminate between infectious and non infectious hepatitis A virus. AB - Hepatitis A virus (HAV) is a major cause of infectious hepatitis worldwide. Detection of HAV in contaminated food or water is a priority research area in laboratories worldwide. Our laboratory has reported previously the development of reverse transcription-polymerase chain reaction (RT-PCR) based detection and typing methods for HAV in contaminated shellfish and produce. It is commonly held that RT-PCR can detect viral genome, but cannot distinguish between infectious and inactivated virus. Therefore, signals obtained after PCR should be considered as false positives unless it can be shown that the sample contains virus capable of infecting a suitable host cell line in culture. We present data to show that this general assumption is not valid. Evidence is provided that demonstrate that signals generated after RT-PCR amplification of viral genome correlated well with the presence of infectious virus in the sample. Viral samples inactivated by heat or UV treatment produced significantly lower signal strength that paralleled infectivity of the sample in cultured cells. The loss of signal strength is most likely the result of damage to the viral RNA that renders it unsuitable for RT PCR. The correlation between PCR signal and infectivity was better following UV inactivation than heat treatment. The procedure may be adapted to other viruses and inactivating agents. PMID- 14738987 TI - An RT-PCR primer pair for the detection of Pospiviroid and its application in surveying ornamental plants for viroids. AB - A primer pair for reverse transcription-polymerase chain reaction (RT-PCR), based on the conserved sequences of the members of genus Pospiviroid was designed to yield a fragment of about 200 base pairs (bp). Since pospiviroids infect a large number of plants species and a few members of the genus Pospiviroid have been already detected in some ornamental plants, the primer pair was evaluated for its efficacy using ornamental plants. The method of return-polyacrylamide gel electrophoresis (R-PAGE) was used to determine the general presence of viroids in the test samples. Efficacy of the primer pair for members of genus Pospiviroid was demonstrated by the detection of Potato spindle tuber viroid (PSTVd) and Tomato chlorotic dwarf viroid (TCDVd) in potato, Chrysanthemum stunt viroid and Iresine viroid in Verbena and Vinca species, and Citrus exocortis viroid in Impatiens species. Specificity of the primer pair became evident, where additional viroids were detected by R-PAGE in Coleus and Magilla species, but they were not amplified by the Pospiviroid primer. This primer pair would be of benefit in indexing ornamental plants in quarantine samples or in viroid-free certification schemes, irrespective of their actual identity. PMID- 14738988 TI - Transformation studies with a human T-cell leukemia virus type 1 molecular clone. AB - In in vitro studies human T-cell leukemia virus type 1 (HTLV-1) may be produced by stable or transient transfection of target cells with an infectious molecular clone. Studies using primary human T cells, the natural targets of HTLV-1 infection, are hampered by difficulty in achieving significant infection with cell-free virus and a poor efficiency of transfection of primary cells. A method is described for the generation of stable cell lines expressing HTLV-1 from an infectious proviral clone. The stably transfected cells can be irradiated and cocultured with human peripheral blood mononuclear cells (PBMC) resulting in infected primary cells. These cells become immortalized, IL-2 dependent lines, which contain integrated copies of provirus and express a full spectrum of viral proteins. Analysis of cellular markers indicates that immortalized cell lines consist of CD3+/CD4+ T cells, matching the most common adult T-cell leukemia (ATL) cell phenotype. The method described has great utility in the study of the replication and transformation capacity of HTLV and HTLV mutant viruses in their natural targets, primary human T lymphocytes. PMID- 14738989 TI - Activation of bovine herpesvirus 4 lytic replication in a non-permissive cell line by overexpression of BoHV-4 immediate early (IE) 2 gene. AB - Bovine herpesvirus 4 (BoHV-4) is a gammaherpesvirus with no clear disease association, it establishes persistent infections in its natural host, the bovine, and in an experimental host, the rabbit. BoHV-4 immediate early 2 (IE2) RNA is the less abundant, spliced, 1.8 kb RNA. The predicted amino acid sequence, of the IE2 protein, reveals that it could encode a 61 kDa protein with amino acid sequence homology to the Epstein-Barr virus (EBV) transactivator R protein and its homologues including, herpesvirus saimiri (HVS), equine herpesvirus 2 (EHV 2), murine herpesvirus 68 and Kaposi's sarcoma-associated herpesvirus (KSHV). We examined recently the interaction of BoHV-4 with a human rhabdomyosarcoma cell line, RD-4, and found that although some infectious viruses can be produced, no cytopathic effect (CPE) was observed [J. Gen. Virol. 81 (2000) 1807]. Because IE2 could play a critical role in BoHV-4 productive infection and its overexpression in RD-4 cells could switch the non-permissive RD-4 status to a permissive one. RD 4 cells expressing stably BoHV-4 IE2 gene were generated. BoHV-4 IE2 induced an increased production of infectious viral particles sufficient to obtain an apparent cytopathic effect. It is concluded that BoHV-4 IE2 is a key factor in determining the outcome of BoHV-4 infection. PMID- 14738990 TI - A simple method for cloning the complete begomovirus genome using the bacteriophage phi29 DNA polymerase. AB - The bacteriophage phiDNA polymerase amplifies circular DNA in a rolling circle amplification mechanism. This characteristic was applied to amplify and clone the complete circular DNA genome of a begomovirus. Total DNA extracted from infected tissue was used as the template of an amplification reaction using the commercial kit TempliPhi (Amersham Biosciences). The amplified DNA could be used for direct sequencing and was cloned after digestion with a single cutting restriction endonuclease. The use of this enzyme simplified the cloning steps and increased the cloning efficiency of the complete genome of a circular plant DNA virus. PMID- 14738991 TI - Chemically modified polysaccharide schizophyllan for antisense oligonucleotides delivery to enhance the cellular uptake efficiency. AB - Schizophyllan is a natural beta-(1-->3)-D-glucan existing as a triple helix in water and as a single chain in dimethylsulfoxide (DMSO), respectively. As we already reported, when some homo-phosphodiester polynucleotide (for example, poly(dA) or poly(C)) is added to the schizophyllan/DMSO solution and subsequently DMSO is exchanged for water, the single chain of schizophyllan forms a complex with the polynucleotide. Furthermore, we have already demonstrated that one of the potential applications of this novel complex is an antisense-oligonucleotide (AS ODN) carrier. This work describes a versatile and universal modification technique which enables us to introduce various functional groups only to the side chain of schizophyllan. This technique consists of periodate oxidation of the glucose side chain (it does not react with the main chain because of the absence of the 1,2-diol group in beta-(1-->3)-glucan) and subsequent introduction of the functional groups into the formyl terminate. In the present work, the introduced functional groups were spermine, octa-arginine (R8), arginine-glycine aspartic acid tripeptide (RGD) and some amino or alpha-amino acid compounds. Using these compounds, we made the complexes and carried out an in vitro antisense assay for them, administrating a phosphorothioate AS ODN to the melanoma A375 or leukemia HL-60 cell lines to depress their c-myb mRNA. When we used the R8 or RGD modified schizophyllan as the antisense carrier, the antisense effect was most enhanced among others. Their superiority can be ascribed to enhancement of endocytosis due to these functional peptides. Furthermore, the cytotoxicity for these two modified schizophyllans was negligibly as small as the natural (unmodified) schizophyllan. One of the peculiar features of our system is that the complex (i.e., carrier+AS ODN) is charged negatively in total, which is different from the conventional systems. The present work has thus clarified that schizophyllan can act as a new potential candidate for AS ODN carriers. PMID- 14738992 TI - Beta-lactoglobulin/polysaccharide interactions during in vitro gastric and pancreatic hydrolysis assessed in dialysis bags of different molecular weight cut offs. AB - The effects of gum arabic, low methylated (LM) pectin or xylan at levels of 0 and 50 wt.% on beta-lactoglobulin (beta-lg) digestibility were studied as well as the interactions between the two macromolecules during in vitro hydrolysis. The proteolysis was performed in a system involving a two-step hydrolysis: either pepsin alone, or pepsin followed by a trypsin/chymotrypsin (T/C) mixture in dialysis bags with molecular weight cut-offs (MWCO) 1000 or 8000 Da. Digestibility was estimated by the N release and by a SDS-PAGE electrophoresis of retentates from the two dialysis bags after hydrolysis. Turbidimetric measurements monitored the structural evolution of mixtures during the two-step hydrolysis. Results showed that beta-lg was almost resistant to peptic digestion and that polysaccharides increased the N release despite a reduction of pepsin activity. This is due to the formation of electrostatic complexes between polysaccharides and beta-lg, which reduced beta-lg aggregation, increasing its solubility. The polysaccharides reduced significantly the beta-lg T/C digestibility as determined using a dialysis bag with a MWCO 1000 Da, without a modification of their enzymatic activities. No significant effect of polysaccharides on the beta-lg digestibility was detected using the dialysis bag with a MWCO 8000 Da. The electrophoresis pattern did not show differences in the profile of retentates in relation with the dialysis bag used. This suggests that non-specific interactions could occur during the second step of hydrolysis between polysaccharides and amino acids or peptides smaller than 8000 Da. PMID- 14738993 TI - A novel water-soluble nanoparticles of hypocrellin B and their interaction with a model protein: C-phycocyanin. AB - A type of novel hypocrellin B gelatin nanoparticles (HB-G-NP), with size of 20 200 nm, was prepared and characterized. The nanoparticles are readily soluble in water or phosphate-buffered saline (PBS). The interaction between HB-G-NP and a fluorescence protein, C-phycocyanin (C-PC) from Spirulina platensis, was studied. It was found that the energy transfer from HB to C-PC was quite efficient, suggesting adsorption of C-PC on surface of the nanoparticles; secondly, the photosensitization of HB resulted in not only the photo-damage of C-PC but also the photobleaching of HB in the presence of oxygen while it did not in the absence of oxygen, suggesting that the movable reactive oxygen species, instead of the immovable anionic radicals of the photosensitizer, should be responsible for the photo-induced processes. Considering the short free diffusion path length of the reactive oxygen species, it can be deduced that smaller or ring-like particles should be more effective for photo-damage of biomolecules or target tissues. PMID- 14738994 TI - Studies on the nature of interaction of iron(III) with alginates. AB - The interactions between the polysaccharide alginate and iron(III) were investigated. The solution properties were studied through pH-metry, viscometry, zeta potential and particle size measurements. In the presence of alginate, iron(III) was stabilized and no precipitation was observed. Studies indicate that iron(III)-alginate system was more stable than iron(III) or alginate alone. The binding constant is of the order of 10(4) M(-1). A case for 'site binding model' for the interaction between alginate and Fe(III) has been made based on the studies using circular dichroism and zeta potential experiments. The number of binding sites per molecule of alginate has been estimated to be 66. This indicates that the alginate can bind more number of Fe(III) ions and thus provide a stable complex which can find wide industrial applications. PMID- 14738995 TI - Food restriction affects energy metabolism in rat liver mitochondria. AB - To examine the effect of 50% food restriction over a period of 3 days on mitochondrial energy metabolism, liver mitochondria were isolated from ad libitum and food-restricted rats. Mitochondrial enzyme activities and oxygen consumption were assessed spectrophotometrically and polarographically. With regard to body weight loss (-5%), food restriction decreased the liver to body mass ratio by 7%. Moreover, in food-restricted rats, liver mitochondria displayed diminished state 3 (-30%), state 4-oligomycin (-26%) and uncoupled state (-24%) respiration rates in the presence of succinate. Furthermore, "top-down" elasticity showed that these decreases were due to an inactivation of reactions involved in substrate oxidation. Therefore, it appears that rats not only adapt to food restriction through simple passive mechanisms, such as liver mass loss, but also through decreased mitochondrial energetic metabolism. PMID- 14738996 TI - Nucleophosmin is a component of the fructoselysine-specific receptor in cell membranes of Mono Mac 6 and U937 monocyte-like cells. AB - The monocyte-like cell lines Mono Mac 6 (MM6) and U937 bind Amadori-modified proteins via fructoselysine (FL)-specific sites with molar masses of 110, 150 and 200 kDa, which can specifically be isolated by an affinity method with magnetobeads coated with glycated polylysine. Using Western blots developed with different anti-nucleophosmin antisera, MS-analysis and immunohistochemistry, we show that the nucleolar protein nucleophosmin is also localized in the cell membrane and is part of the 150- and 200-kDa membrane protein fractions of FL specific binding membrane proteins. This is the first evidence that nucleophosmin is not only existing in the nucleolus and cytoplasm, but also, like nucleolin, is in the cell membrane. PMID- 14738997 TI - Low molecular weight chitosans: preparation with the aid of papain and characterization. AB - Low molecular weight chitosans (LMWC) of different molecular weight (4.1-5.6 kDa) were obtained by the depolymerization of chitosan using papain (from Carica papaya latex, EC. 3.4.22.2) at optimum conditions of pH 3.5 and 37 degrees C for 1-5 h. Scanning electron microscopy (SEM) showed approximately 15-fold decrease in the particle size after depolymerization. Decrease in the molecular weight was associated with decrease in the degree of acetylation (DA) as evidenced by circular dichroism (CD), FT-IR and solid-state CP-MAS 13C-NMR data. X-ray diffraction pattern revealed slight decrease in the crystallinity index (CrI) whereas the 13C-NMR data showed molecular inhomogeneity. LMWC showed lytic effect towards Bacillus cereus and Escherichia coli more efficiently than native chitosan. The growth inhibitory effect was maximal towards B. cereus, with minimum inhibitory concentration (MIC) of 0.01% (w/v). PMID- 14738998 TI - Potentiation of beta-amyloid polymerisation by low-density lipoprotein enhances the peptide's vasoactivity. AB - Alzheimer's disease (AD) is characterised by the accumulation of insoluble beta amyloid (A beta) fibrils in the brain. Factors that promote A beta fibrillogenesis may influence the pathogenesis of AD and represent targets for therapeutic intervention. Some A beta deposited in AD may originate in the circulation and plasma factors could promote A beta deposition, particularly in the cerebrovasculature. We investigated the effects of plasma low-density lipoprotein (LDL), in both its native and oxidised forms, on A beta(1-40) fibrillogenesis and vasoactivity. LDL enhanced A beta fibrillogenesis in a process dependent on LDL concentration and the oxidative state of the lipoprotein, as indicated by measurements of thiobarbituric acid reactive substances (TBARS) and conjugated dienes. LDL's actions were inhibited by the iA beta 5 peptide, suggesting that LDL-induced A beta polymerisation involved beta pleated sheet formation. Potentiated A beta polymerisation was reflected by enhanced A beta-mediated vascular responses. Human endothelial cells exposed to fibrillar A beta generated with LDL, especially oxidised LDL, exhibited decreased 20S proteasome activity. Rat aortic ring constriction induced by noradrenaline was enhanced by A beta fibrils generated with LDL, with oxidised LDL producing the more marked effects. Should plasma lipoproteins prove to play a role in cerebral A beta deposition their modification with statins or antioxidants may offer therapeutic benefit. PMID- 14738999 TI - Expression of human arginine decarboxylase, the biosynthetic enzyme for agmatine. AB - Agmatine, an amine formed by decarboxylation of L-arginine by arginine decarboxylase (ADC), has been recently discovered in mammalian brain and other tissues. While the cloning and sequencing of ADC from plant and bacteria have been reported extensively, the structure of mammalian enzyme is not known. Using homology screening approach, we have identified a human cDNA clone that exhibits ADC activity when expressed in COS-7 cells. The cDNA and deduced amino acid sequence of this human ADC clone is distinct from ADC of other forms. Human ADC is a 460-amino acid protein that shows about 48% identity to mammalian ornithine decarboxylase (ODC) but has no ODC activity. While naive COS-7 cells do not make agmatine, these cells are able to produce agmatine, as measured by HPLC, when transfected with ADC cDNA. Northern blot analysis using the cDNA probe indicated the expression of ADC message in selective human brain regions and other human tissues. PMID- 14739000 TI - Hemispheric specialization for language. AB - Hemispheric specialization for language is one of the most robust findings of cognitive neuroscience. In this review, we first present the main hypotheses about the origins of this important aspect of brain organization. These theories are based in part on the main approaches to hemispheric specialization: studies of aphasia, anatomical asymmetries and, nowadays, neuroimaging. All these approaches uncovered a large inter-individual variability which became the bulk of research on hemispheric specialization. This is why, in a second part of the review, we present the main facts about inter-individual variability, trying to relate findings to the theories presented in the first part. This review focuses on neuroimaging as it has recently given important results, thanks to investigations of both anatomical and functional asymmetries in healthy subjects. Such investigations have confirmed that left-handers, especially "familial left handers", are more likely to have an atypical pattern of hemispheric specialization for language. Differences between men and women seem less evident although a less marked hemispheric specialization for language was depicted in women. As for the supposed relationship between anatomical and functional asymmetries, it has been shown that the size of the left (not the right) planum temporale could explain part of the variability of left hemispheric specialization for language comprehension. Taken as a whole, findings seem to vary with language tasks and brain regions, therefore showing that hemispheric specialization for language is multi-dimensional. This is not accounted for in the existing models of hemispheric specialization. PMID- 14739001 TI - A potentially critical role of phospholipases in central nervous system ischemic, traumatic, and neurodegenerative disorders. AB - Phospholipases are a diverse group of enzymes whose activation may be responsible for the development of injury following insult to the brain. Amongst the numerous isoforms of phospholipase proteins expressed in mammals are 19 different phospholipase A2's (PLA2s), classified functionally as either secretory, calcium dependent, or calcium independent, 11 isozymes belonging to three structural groups of PLC, and 3 PLD gene products. Many of these phospholipases have been identified in selected brain regions. Under normal conditions, these enzymes regulate the turnover of free fatty acids (FFAs) in membrane phospholipids affecting membrane stability, fluidity, and transport processes. The measurement of free fatty acids thus provides a convenient method to follow phospholipase activity and their regulation. Phospholipase activity is also responsible for the generation of an extensive list of intracellular messengers including arachidonic acid metabolites. Phospholipases are regulated by many factors including selective phosphorylation, intracellular calcium and pH. However, under abnormal conditions, excessive phospholipase activation, along with a decreased ability to resynthesize membrane phospholipids, can lead to the generation of free radicals, excitotoxicity, mitochondrial dysfunction, and apoptosis/necrosis. This review evaluates the critical contribution of the various phospholipases to brain injury following ischemia and trauma and in neurodegenerative diseases. PMID- 14739002 TI - An overview of new pharmacological treatments for cerebrovascular dysfunction after experimental subarachnoid hemorrhage. AB - Cerebral vasospasm and the resulting cerebral ischemia occurring after subarachnoid hemorrhage (SAH) are still responsible for the considerable morbidity and mortality in patients affected by cerebral aneurysms. Mechanisms contributing to the development of vasospasm, abnormal reactivity of cerebral arteries and cerebral ischemia after SAH have been intensively investigated in recent years. It has been suggested that the pathogenesis of vasospasm is related to a number of pathological processes, including endothelial damage, smooth muscle cell contraction resulting from spasmogenic substances generated during lyses of subarachnoid blood clots, changes in vascular responsiveness and inflammatory or immunological reactions of the vascular wall. A great deal of experimental and clinical research has been conducted in an effort to find ways to prevent these complications. However, to date, the main therapeutic interventions remain elusive and are limited to the manipulation of systemic blood pressure, alteration of blood volume or viscosity, and control of arterial dioxide tension. Even though no single pharmacological agent or treatment protocol has been identified which could prevent or reverse these deadly complications, a number of promising drugs have been investigated. Among these is the hormone erythropoietin (EPO), the main regulator of erythropoiesis. It has recently been found that EPO produces a neuroprotective action during experimental SAH when its recombinant form (rHuEPO) is systemically administered. This topic review collects the relevant literature on the main investigative therapies for cerebrovascular dysfunction after aneurysmal SAH. In addition, it points out rHuEPO, which may hold promise in future clinical trials to prevent the occurrence of vasospasm and cerebral ischemia after SAH. PMID- 14739003 TI - Fas ligand/Fas system in the brain: regulator of immune and apoptotic responses. AB - Apoptosis, also known as programmed cell death, is the major type of cell death involved in normal development, regeneration, proliferation and pathologic degeneration in the central nervous system (CNS). The apoptotic process can be divided further into two pathways depending on the involvement of mitochondria and related biochemical cascades. The internal pathway of apoptosis is initiated by a variety of cytotoxic stimuli and mediated by the release of cytochrome c and subsequent activation of downstream caspases. The external pathway is mainly triggered by ligation of death receptors such as Fas, tumor necrosis factor (TNF) related apoptosis inducing ligand-R1 (TRAIL-R1), TRAIL-R2 and TNFRp55, and mediated by direct activation of upstream caspases. The Fas-FasL system has been known as a prototypic inducer of extrinsic cell death responsible for cell mediated cytotoxicity, peripheral immune regulation, immune privilege and "counterattack" of malignant tumor cells against the host immune system. Fas and FasL are expressed in the normal CNS, and expression increases in inflamed and degenerated brains. Like other specialized tissues such as the eye and testis, the Fas-FasL system is thought to be involved in immune suppressed status in the CNS. Expression of Fas and FasL is significantly elevated in a variety of the neurologic disorders, suggesting the possibility that this system may play roles in degenerative and inflammatory responses in the CNS. Therefore, the FasL-Fas system should be considered as a double-edged sword in the CNS: maintaining the immune suppressed status in normal brain and inducing neuronal cell death and inflammation in a variety of neurologic disorders. PMID- 14739004 TI - Visual categorization and the inferior temporal cortex. AB - We investigated the effects of categorization on the representation of stimulus features in combined psychophysical-electrophysiological experiments. We used parameterized line drawings of faces and fish as stimuli, and we varied the relevance of the different features for the categorization task. The psychophysical and electrophysiological data support an exemplar-based framework for visual object recognition. We recorded from visual neurons in the anterior inferior temporal (IT) cortex of macaque monkeys, while they were performing a categorization task. The visual neurons did not respond selectively to one stimulus set, or to one category. The majority of the anterior IT feature selective neurons were tuned for features that were diagnostic for the categorization task. We argue that this fine-tuning of the neurons reflects the perceptual sensitization to the diagnostic features. PMID- 14739005 TI - Haloperidol versus risperidone on rat "early onset" vacuous chewing. AB - Similarly to acute rat catalepsy, "early onset" vacuous chewing movements (VCMs) induced by subchronic treatment with antipsychotic have recently been proposed as a model of human extrapyramidal symptoms. In the present study, the propensities of haloperidol and risperidone in inducing rat "early onset" VCMs were compared using doses of the two antipsychotics that acutely induce similar catalepsy. Comparable rat catalepsy states were observed when the effects produced by 0.1, 0.5, and 1mg/kg of haloperidol were compared with those induced by 1, 4, and 10mg/kg of risperidone, respectively. These doses of the two antipsychotics were then administered twice a day for 4 weeks and VCMs scored after 12h, 5 days, or 3 weeks of drug withdrawal. Among the haloperidol-treated groups, only those rats injected with 0.5 and 1mg/kg showed high levels of VCMs after 12h and 5 days of drug withdrawal when compared to vehicle-treated rats, while basal levels of VCMs were reached after 3 weeks from the last injection. High VCMs levels were observed in risperidone-treated rats only at the dose of 10mg/kg and after 12h of drug withdrawal, but not after 5 days or 3 weeks. The present results indicated that haloperidol possessed a much higher propensity to induce rat "early onset" VCMs than risperidone. PMID- 14739006 TI - The role of hippocampal signaling cascades in consolidation of fear memory. AB - We investigated the involvement of hippocampal protein kinase A (PKA), protein kinase C (PKC), calcium/calmodulin-dependent protein kinases (CaMK II), and mitogen-activated extracellular signal-regulated kinases 1 and 2 (Mek-1/2) in the phosphorylation of their downstream targets extracellular signal-regulated kinases 1 and 2 (Erk-1/2), cAMP-response element-binding protein (CREB), Elk-1, and p90 ribosomal S6 kinase 1 (p90Rsk-1). The role of these processes in memory consolidation of conditioned fear was determined. C57BL/6N mice were injected into the dorsal hippocampus with inhibitors of PKA, PKC, CaMK II, Mek-1/2, or vehicle before training consisting of a single exposure to a context, tone, and footshock. Freezing behavior of mice reflecting fear memory was scored after their re-exposure to the conditioned stimuli. Inhibition of PKA impaired context- and tone-dependent fear conditioning and significantly reduced the phosphorylation of Elk-1, p90Rsk-1, Erk-1/2, and CREB. PKC inhibition also impaired context- and tone-dependent fear conditioning and prevented the phosphorylation of Erk-1/2, Elk-1, and CREB, without affecting p90Rsk-1. Inhibition of CaMK II did not affect fear conditioning and reduced the phosphorylation of Erk-1/2, Elk-1, CREB, and p90Rsk-1 only transiently, whereas Mek-1/2 inhibition was ineffective in all experiments. It was concluded that hippocampal PKA and PKC play crucial roles in one-trial fear conditioning. Erk 1/2, Elk-1, and CREB were identified as common targets of PKA, PKC, and CaMK II during memory consolidation, however, the time window and sequence of their phosphorylation was specific for the individual kinase. PMID- 14739007 TI - Increased amphetamine-induced locomotion during inactivation of the basolateral amygdala. AB - At low doses, amphetamine has been shown to produce reliable increases in locomotor activity through its actions on the mesolimbic dopamine (DA) terminals in the nucleus accumbens (NAC). The basolateral amygdala (BLA) has recently been reported to have anatomical projections to the NAC, suggesting that it might serve to alter or modulate the function of the NAC. To test this hypothesis, the current experiment produced lidocaine-reversible lesions of the BLA and assessed changes in NAC function by examining alterations in locomotor activity in response to s.c. amphetamine (2mg/kg). While BLA inactivation alone was found to have no effect on spontaneous or basal locomotor activity, it produced a significant potentiation of amphetamine-induced hyperactivity. These results suggest that BLA inactivation removes a system that inhibits the locomotor response to amphetamine. The data are, therefore, consistent with the view that the BLA may serve to modulate NAC function. PMID- 14739008 TI - Visually guided locomotion in the gerbil: a comparison of open- and closed-loop control. AB - Two experiments were conducted in order to explore the effects of visual feedback on control of locomotion in the gerbil. In the first experiment, gerbils were trained to run down an alleyway towards a visual target in order to obtain food reward. One group of animals was trained to run to a target whose size never varied while another group was trained to run to a target whose size varied randomly from trial to trial. On some trials, the target's size was changed dynamically during running to determine whether gerbils used this dynamic size change information to compute time to collision (tau). Results suggested that neither group used retinal image size information but both groups seemed able to compute tau. In a second experiment, gerbils were trained in the same way as in experiment 1, but on some trials the target was extinguished during the run. In this condition, probe trials showed that gerbils used retinal image size to compute target distance. Collectively, the results showed that gerbils were able to rapidly and flexibly utilize available information to complete a visually guided running task. The results are discussed in the context of the psychophysics of cue combination and its neural underpinnings. PMID- 14739009 TI - Cognitive deficits in adult rats by lead intoxication are related with regional specific inhibition of cNOS. AB - It is well known that lead can affect several cognitive abilities in developing animals. In this work, we investigate the effects of different sub-chronic lead doses (0, 65, 125, 250 and 500 ppm of lead acetate in their drinking water for 14 days) in the performance of male adult rats in a water maze, cue maze and inhibitory avoidance tasks. We found that the acquisition of these tasks was not affected by lead, however, the highest dosage of lead (500 ppm) impaired memory consolidation in spatial and inhibitory avoidance tasks, but not in cue maze task while the 250 ppm dose only affected retrieval of spatial memory. Additionally, hippocampal long-term potentiation (LTP) induction in the perforant path after exposing adult rats to different doses of lead was studied. LTP induction was affected in a dose-dependent manner, and treatments of 250 and 500 ppm completely blocked LTP. We investigated the effects of lead intoxication on the activity of constitutive nitric oxide synthase (cNOS) in different brain regions of adult animals. The activity of cNOS was significantly inhibited in the hippocampus and cerebellum but not in the frontal cortex and brain stem, although lead had accumulated in all brain regions. These results suggest that lead intoxication can impair memory in adult animals and this impairment might be related with region-specific effects on cNOS activity. PMID- 14739010 TI - Reversible lesion of the rat's orbitofrontal cortex interferes with hippocampus dependent spatial memory. AB - In this study, tetrodotoxin (TTX) inactivation was employed to evaluate the involvement of the rat's orbitofrontal cortex (OFC) in hippocampus-dependent spatial memory using Morris water maze (MWM) and place avoidance learning (PAL) tasks. In Experiment 1, rats trained in MWM task with two blocks of four trials per day for 3 consecutive days received bilateral injections of either TTX or saline into the OFC 60 min before each daily training session. The acquisition of spatial memory was evaluated 24h after the last training day and it was shown an impairment by the TTX. In Experiment 2, bilateral intra-OFC injections of TTX or saline were made immediately after two blocks of four trials. Testing 24h later, it was revealed that TTX also impairs spatial memory consolidation. In Experiments 3 and 4, rats were trained in a single 30-min session to avoid a 60 degrees segment of the stable circular (80-cm diameter) arena, entering which was punished by a mild shock (PAL task) and retention was tested 24h later in a 30 min extinction session. Bilateral injections of TTX or saline were made into the OFC 60 min before training or immediately after training. Again, TTX impaired the place avoidance retention when it was injected into the OFC either before (acquisition phase) or after (consolidation phase) training. These findings indicate that functional integrity of the OFC is necessary for both the acquisition and the consolidation of hippocampus-dependent spatial memory in rats. PMID- 14739011 TI - Experience and cortical control over the pubertal transition to rougher play fighting in rats. AB - With the onset of puberty, play fighting in rats decreases in frequency and the tactics of attack and defense that are used are rougher. Previous studies have shown that the changes in the frequency of play and in the use of defensive tactics arise independently of social experience. Furthermore, while the former involves subcortical regulation, the latter depends on cortical mechanisms. In this study, the possible mechanisms regulating the developmental changes in the tactics of attack were examined. Two experiments were conducted using male rats. In the first study, rats reared in isolation from weaning were compared to rats reared in pairs, and were tested in the juvenile and early post-pubertal phases (30 and 60 days postnatally). In the second experiment, rats with their cortex removed shortly after birth were compared to sham-treated controls, and were tested in pairs at both the juvenile and early adult phases (30 and 90 days). Two measures of 'roughness,' derived from previous studies, were measured. Results showed that isolation-reared rats had the typical age-related changes in roughness of attack, whereas decorticated rats failed to show this age-related modulation, maintaining, or even exaggerating, the juvenile-typical pattern of attack. These findings suggest that social experience is not needed for this developmental change to occur, and that an intact cortex is needed to regulate this change in behavior. PMID- 14739012 TI - The non-steroidal anti-inflammatory drug diclofenac sodium attenuates lipopolysaccharide-induced alterations to reward behavior and corticosterone release. AB - Non-steroidal anti-inflammatory drugs (NSAIDs) have been shown to counteract stress hormone and pro-inflammatory cytokine activation. To extend these findings, we tested whether the NSAID diclofenac sodium would attenuate lipopolysaccharide (LPS)-induced reductions in reward behavior. In the first experiment, male, Wistar rats pressed a lever for food reward and subsequently received 10 days treatment of saline (1 ml/kg, s.c.) or diclofenac (2.5mg/kg, s.c.). On the subsequent test day, rats were given a final injection of saline or diclofenac 30 min prior to LPS (20 micrograms/kg, i.p.). LPS significantly reduced rate of food self-administration and total reinforcers obtained and increased corticosterone levels in saline-treated rats, while these effects were significantly attenuated in diclofenac-treated rats. In the second experiment, rats pressed a lever for sweetened milk. In contrast to food self-administration, acute LPS exposure did not reduce rate of responding or total reinforcers obtained in either saline- or diclofenac-treated rats. In the third experiment, rats trained to press a lever for sweetened milk were pre-exposed to a high dose of LPS (250 micrograms/kg, i.p.) 2 weeks prior to a challenge injection of LPS. In this case, LPS challenge significantly reduced rate of sweetened milk self administration, but not total reinforcers obtained, in saline-treated rats. Rats treated with diclofenac did not exhibit reductions in rate of responding or total reinforcers obtained. Overall, the data indicate that the NSAID diclofenac sodium counteracts LPS-induced reductions in reward behavior and corticosterone release, and may therefore have therapeutic potential for specific components of endotoxin induced sickness behavior, including anhedonia. PMID- 14739013 TI - Stress alters caffeine action on investigatory behaviour and behavioural inhibition in the mouse. AB - A lot of studies have demonstrated that the physiological action of drugs can be modified by stressors. The present study investigates the effects of stressors on caffeine action on investigatory behaviour and behavioural inhibition in C57Bl6/J mice. For 6 consecutive days, the mice were subjected to one stressful procedure each day consisting on days 1 and 3 of immersion in cold water for 10 periods of 10s each, on days 2 and 5 of a restraint stress for 2 periods of 5 min each, and on days 3 and 6 of placing the animals in a lit openfield for a 10-min period. Saline or caffeine (30, 60 or 120 mgkg-1) were i.p. administered immediately after the last stressful experience, and the animals were tested behaviourally 10 min later. Their behaviour was compared to that of unstressed mice injected with either saline or caffeine. The results show that: (1) in saline-treated mice, stress decreased the investigatory behaviour and increased the behavioural inhibition; (2) in unstressed mice, caffeine decreased the investigatory behaviour in a dose-dependent manner; moreover, behavioural disinhibition, which appeared at low doses of the drug, did not at higher doses; (3) in stressed mice, the dose-dependent action of caffeine was almost abolished and the drug elicited, whatever the dose, a slight increase of the investigatory behaviour and a strong behavioural disinhibition. It is concluded that stress antagonizes the inhibitory action of caffeine on the investigatory behaviour and potentiates its action on behavioural disinhibition. The results are discussed in terms of interaction of stress and caffeine on the dopaminergic system. PMID- 14739014 TI - Chronic aerial exposure to glucorticoids or beta-agonists affects avoidance learning and exploratory motivation in rats. AB - The purpose of the present work was to examine if the conventional asthma treatments in humans (inhalation of glucocorticoids or beta-agonists, administered in a chronic regimen) might affect behavioral processes (learning and exploratory motivation) in rats. Adult male rats were exposed to an atmosphere saturated with either saline, budesonide (a glucocorticoid), or salbutamol (a beta-adrenergic receptor agonist) in a forced ventilation cage, connected to a nebulizer for 5 min twice a day for 15 days at the same hours of the day. Doses of budesonide in the nebulizing solution were 0.116, 1.16, and 11.6mM. Doses of salbutamol in the nebulizing solution were 1.3, 13, and 130 mM. Forty-eight hours after treatment, the different groups were subjected to exploration of an elevated asymmetric plus-maze (APM, model of exploratory motivation), and 24h later to learning of an avoidance response to an ultrasonic tone in a two-compartment cage (model of memory and learning). Results showed that budesonide induces moderate effects on exploratory motivation. In one of the fear-inducing arms (single wall arm), exploration decreased and this effect was not dose dependent. In the cognitive model, glucocorticoids affected slightly the latency to escape but with no interference in memory efficiency. On the other hand, at the lower dose in the APM, salbutamol increased significantly the exploration of both fear-inducing arms (no walls and single wall arms). In the learning model, the beta-agonist induced two opposing effects. The lower dose (1.3mM) facilitated learning and the higher dose (13 mM) inhibited learning instead. In conclusion, results are compatible with the notion that inhaled glucocorticoids or beta-agonists might cross the lung aerial barrier into the blood compartment, exerting effects on learning and motivation functions. PMID- 14739015 TI - Development of configural 3D object recognition. AB - There is evidence for the late development in humans of configural face and animal recognition. We show that the recognition of artificial three-dimensional (3D) objects from part configurations develops similarly late. We also demonstrate that the cross-modal integration of object information reinforces the development of configural recognition more than the intra-modal integration does. Multimodal object representations in the brain may therefore play a role in configural object recognition. PMID- 14739016 TI - Rapid screening method for Schistosoma japonicum infection using questionnaires in flood area of the People's Republic of China. AB - A rapid, simple, and effective method using questionnaires is described for screening of high-risk individuals of schistosomiasis japonica after the occurrence of major flooding. A case-control study design was adopted in two randomly selected villages from Dongting Lake area in the People's Republic of China that are endemic for Schistosoma japonicum. Information about contagious water exposure history and several other risk factors were collected retrospectively from 246 individuals, aged 9-60 years. A probability model was developed by logistic regression analysis, which included six variables, namely (i) knowledge of Schistosoma transmission, (ii) education attainment, (iii) annual income before flood year per person, (iv) duration of contagious water exposure due to swimming and paddling, (v) intensity of contagious water exposure due to occupational activities, and (vi) duration of contagious water exposure due to recreational activities. The receiver-operating characteristic (ROC) curve was plotted and the sensitivity and specificity were calculated. The internal consistency of the probability model was good. The area below the ROC curve was 0.90. If the probability cutoff value of diagnosing an infection with S. japonicum was defined as 0.35, both sensitivity and specificity were above 82%, whereas positive and negative predictive values were 70 and 91%, respectively. We conclude that questionnaires are a viable tool for screening of high-risk individuals of S. japonicum infections in lake communities of China after flooding occurred, opening new avenues for schistosomiasis control. PMID- 14739017 TI - Visceral leishmaniasis in immunocompromised patients with and without AIDS: a comparison of clinical features and prognosis. AB - Visceral leishmaniasis is basically a disease of healthy infants and adults. However, in the last decade an increasing number of cases of kala azar in immunocompromised patients have been reported with emphasis on atypical manifestations of the disease. During a period of 11 years, 20 immunocompromised patients with AIDS (12 patient), haematological neoplasia (3 patients), corticosteroid therapy (3 patients) or renal transplantation (2 patients) were studied by one or more of the authors. We did not find differences in the presentation of leishmaniasis between patient with or without AIDS and most patients had fever, enlargement of the liver and spleen, blood cytopenias and biochemical abnormalities. Serology was more frequently positive in HIV-negative than in HIV-positive patients (100% versus 63.6%; P=0.13). Bone marrow biopsy was diagnostic in 66% and 87% of patients with and without AIDS, respectively. Failure of anti-leishmanial therapy occurred in 6 of 19 patients treated (31.5%), and 3 patients with AIDS and another 3 without AIDS died during the first episode of leishmaniasis. Of 12 survivors, relapses occurred in five (41.6%). Only patients in whom immunosuppression was ameliorated by means of antiretroviral therapy or by reduction of corticosteroid and other immunosuppressive drugs did not relapse. Treatment of kala azar in immunocompromised host is in satisfactory and new drugs or strategies are urgently needed. PMID- 14739018 TI - Which children come to the health centre for treatment of malaria? AB - Malaria remains one of the major public health problems for children in Guinea Bissau and the major reason for using the health services. Little is known about factors with impact on whether the mothers seek malaria treatment for their children at a health centre. Mothers of children coming to the Bandim Health Centre in Bissau and age-matched controls selected from the registration system of the Bandim Health Project were interviewed about treatment habits, socio economic standards and other factors associated with the mother seeking treatment for her child at a health centre. In a multivariate analysis, the controls were more often found to have chloroquine at home and had more often taken home treatment for suspected malaria. On average 2.97 persons among cases shared bed as opposed to only 2.02 among controls, bed-crowding thus, being a factor determining health seeking behaviour (P=0.0001). Low socio-economic status, as measured by the non-availability of electricity, was also a significant predicting factor (P=0.02). The possibilities of promoting home treatment should be further evaluated, not only to avoid unnecessary visits to the professional health system, but also to decrease the morbidity from acute malaria. PMID- 14739019 TI - Human transportation influences Aedes aegypti gene flow in Southeast Asia. AB - Estimates of population genetic organization and gene flow of Aedes aegypti, the main vector of dengue viruses, have provided insights into dengue epidemiology. To assess the role of the vector in the changing pattern of dengue in Southeast Asia, extensive studies on the genetic differentiation of Aedes aegypti have been carried out. Among them, vector differentiation has been estimated for Cambodia, Thailand and South Vietnam using microsatellite markers. Analysis showed that there was less genetic differentiation between mosquito populations from Ho Chi Minh City and Phnom Penh than from either of them and Thailand, suggesting that passive migrations through human transportation help to explain this pattern of differentiation. PMID- 14739020 TI - PCR-RFLP to identify Leishmania (Viannia) braziliensis and L. (Leishmania) amazonensis causing American cutaneous leishmaniasis. AB - A PCR-RFLP based method was developed to diagnose and identify the Leishmania species causing American cutaneous leishmaniasis (ACL) in a panel of clinical samples obtained from an endemic region of Brazil. The comparison of the results obtained by PCR-RFLP and PCR-hybridization in the identification of Leishmania (Viannia) braziliensis and L. (Leishmania) amazonensis were highly concordant (kappa=91.5%). The PCR-RFLP method was reliable, fast and easy to conduct on biopsies and presents potential value of utmost importance for the diagnosis and identification of Leishmania in clinical specimens, infected reservoirs and vectors. PMID- 14739021 TI - Microsatellite DNA polymorphism and heterozygosity in the malaria vector mosquito Anopheles funestus (Diptera: Culicidae) in east and southern Africa. AB - There has been an increase in malaria cases in southern African countries in recent years due to the presence of populations of Anopheles funestus that are resistant to the pyrethroid class of insecticides. Since A. funestus is one of the major African malaria vectors, knowledge of its genetic structure will benefit control strategies, such as the management of insecticide resistance, by allowing predictions to be made of possible spread of the resistance. This study uses microsatellite DNA markers to analyze samples from five countries in east (Kenya and Uganda), central (Malawi) and southern (South Africa and Mozambique) Africa. There were deviations from Hardy-Weinberg expectations for some loci in all population samples but this was probably due to the presence of null alleles. High levels of genetic diversity were observed (mean alleles per locus = 6.5-10; unbiased H=0.23-0.89). Low differentiation was observed between Kenya and Uganda (average F(ST)=0.002, R(ST)=0.0001) and between Mozambique and South Africa (F(ST)=0.0004, R(ST)=0.02), contrary to high differentiation among the central and southern Africa samples (average F(ST)=0.023, R(ST)=0.027). High differentiation was measured across the region (mean F(ST)=0.04, R(ST)=0.08), east versus Malawi (F(ST)=0.067, R(ST)=0.089) or southern Africa populations (F(ST)=0.068, R(ST)=0.15). A test of isolation by distance along the east-central south transect gave evidence (R(2)=0.50, P<0.001) that geographic distance limits gene flow in A. funestus. PMID- 14739022 TI - Eosinophilic meningitis in a child raising snails as pets. AB - The rat lungworm (Angiostrongylus cantonensis) is the principal cause of eosinophilic meningitis or meningoencephalitis worldwide. It is endemic in Taiwan and the Asia Pacific area. We report the case of a 10-year-old boy who was referred to us suffering from intermittent headache, low-grade fever and blurred vision of 4-5 days' duration. He had been treated for gastroenteritis just prior to referral. The patient's history was unremarkable, except that he raised snail (Ampullarium canaliculatus) as pet at home. On physical examination, the patient's consciousness was alert and well oriented. No papilledema was found on fundal examination. The neurological examination revealed normal cranial nerve function, mild weakness of both lower limbs and normal deep tendon reflexes, but positive Babinski and Kernig signs. Laboratory findings showed peripheral eosinophilia, elevated immunoglobulin E level, cerebrospinal fluid eosinophilic pleocytosis and the presence of stage 3 A. cantonensis larvae, which confirmed the diagnosis of eosinophilic meningitis. A 2-week course of mebendazole combined the glucocorticosteroids was beneficial in relieving headache, paresthesia and the other eosinophilic meningitis symptoms in the patient. PMID- 14739023 TI - In vitro activity of quinolines against Plasmodium falciparum in Gabon. AB - The assessment of drug sensitivity of Plasmodium falciparum to antimalarial drugs is of vital interest for malaria endemic regions. We conducted a follow-up study to monitor the in vitro activity of the most commonly used quinolines against fresh P. falciparum isolates in Lambarene, Gabon by measuring schizont maturation inhibition in 2002. Mean 50% effective concentration levels for chloroquine, quinine, and mefloquine were 5.5micromol/l blood, 286nmol/l blood medium mixture (BMM), and 1.1micromol/l blood, respectively. All isolates (n=40) were found to be highly resistant to chloroquine. One isolate was resistant to mefloquine and five isolates were presenting borderline-resistance. All isolates were inhibited by quinine concentrations below the threshold of resistance (n=43). Besides the observation of an increasing number of borderline resistant isolates to mefloquine, an extremly high parasite resistance to chloroquine-still officially the first line antimalarial in Gabon-seems to be of particular concern. PMID- 14739024 TI - High prevalence of Plamodium malariae infections in a Brazilian Amazon endemic area (Apiacas-Mato Grosso State) as detected by polymerase chain reaction. AB - Plasmodium malariae is commonly confounded with Plasmodium vivax at the microscopic examination of thick blood smear. In the present study, we used a nested PCR assay to amplify a species-specific sequence of the 18S SSU rRNA gene of Plasmodium in blood samples of 497 individuals living in an endemic region of the Brazilian Amazon basin. We have found that, while the microscopic examination of thick blood smears showed a P. malariae prevalence of 1.2% (6 out of 497), the nested PCR revealed 11.9% (59 out of 497) of positive cases for this specie. These results point to the need of the development or use of a more accurate diagnosis method to distinguish between P. malariae and P. vivax, which is particularly important in view of the fact that the choice of drug for the antimalarial therapy depends on the parasite species. PMID- 14739025 TI - Proinflammatory and anti-inflammatory cytokines in pregnant women chronically infected with Trypanosoma cruzi. AB - Mother-to-child transmission of intracellular parasites could be related to the production of immunoregulatory cytokines. The levels of gamma interferon (IFN gamma), tumor necrosis factor-alpha (TNF-alpha) and lnterleukin (IL)-10 were evaluated during pregnancy in sera of women chronically infected with Trypanosoma cruzi that delivered infected or non-infected children. The levels of IL-10 increased in both, women only pregnant and only infected, compared to non infected non-pregnant women. However, in pregnant women chronically infected with T. cruzi, IL-10 did not increase significantly, neither in the mothers of infected nor in the mothers of non-infected children. The levels of the inflammatory cytokine TNF-alpha were not affected in normal pregnancy but increased in the infected mothers of non-infected children. The levels of IFN gamma did not increase in the groups studied, indicating that the production of this pro-inflammatory cytokine was controlled, even when the levels of IL-10 did not increase, as in pregnant women chronically infected with T. cruzi. PMID- 14739026 TI - Mapping the potential distribution of Phlebotomus martini and P. orientalis (Diptera: Psychodidae), vectors of kala-azar in East Africa by use of geographic information systems. AB - The distribution of two principal vectors of kala-azar in East Africa, Phlebotomus martini and Phlebotomus orientalis were analysed using geographic information system (GIS) based on (1) earth observing satellite sensor data: Normalized Difference Vegetation Index (NDVI) and midday Land Surface Temperature (LST) derived from advanced very high resolution radiometer (AVHRR) of the global land 1km project of United States Geological Survey (USGS), (2) agroclimatic data from the FAO Crop Production System Zone (CPSZ) of the Intergovernmental Authority on Development (IGAD) sub-region, and (3) the FAO 1998 soils digital map for the IGAD sub-region. The aim was to produce a predictive risk model for the two vectors. Data used for the analysis were based on presence and absence of the two species from previous survey collections in the region (mainly Ethiopia, Kenya and Somalia). Annual, wet season and dry season models were constructed. Although all models resulted in more than 85% positive predictive values for both species, the best fit for the distribution of P. martini was the dry season composite (NDVI 0.07-0.38 and LST 22-33 degrees C) with a predictive value of 93.8%, and the best fit for P. orientalis was the wet season composite (NDVI 0.01 to 0.34 and LST 23-34 degrees C) with a predictive value of 96.3%. The two seasonal composites models derived from satellite data were largely similar with best fit models developed based on the CPSZ climate data: average altitude (12 1900m), average annual mean temperature (15-30 degrees C), annual rainfall (274 1212mm), average annual potential evapotranspiration (1264-1938mm) and readily available soil moisture (62-113mm) for P. martini; and average altitude (200 2200m), annual rainfall (180-1050mm), annual mean temperature (16-36 degrees C) and readily available soil moisture (67-108mm) for P. orientalis. Logistic regression analysis indicated LST dry season composite of the satellite data, average altitude, mean annual temperature and readily available soil moisture of the CPSZ data as the best ecological determinants for P. martini while LST annual composite was the only important ecological determinant for P. orientalis. Spearman's rank correlation revealed several factors to be important determinants for the distribution of the two vectors. None of the soil types analysed appeared to be important determinant for the two species in East Africa, unlike in Sudan where P. orientalis is mainly associated with eutric vertisol (black cotton clay soil). PMID- 14739027 TI - Dengue virus infection in travellers returning to Berlin, Germany: clinical, laboratory, and diagnostic aspects. AB - BACKGROUND: Dengue is a mosquito-borne viral infection endemic throughout the tropics and subtropics. The global prevalence of dengue has grown dramatically in recent years and it has been recognized as a potential hazard to tourists. OBJECTIVE: In this study, we analyzed the epidemiology, clinical manifestations, laboratory features and serological/virological results in a series of German travellers returning to Berlin with acute dengue virus infection. STUDY DESIGN: Laboratory-confirmed dengue virus infections among German travellers returning to Berlin were studied retrospectively during the period of 1993-2001. Seventy-one patients tested positive for dengue fever and were included in this study. RESULTS: The majority of patients (77.5%) contracted the disease in South Central and South East Asia. The most important clinical characteristics were fever and prostration (100%), headache, predominantly frontal or retroorbital (86%), arthralgia (79%), morbilliform rash (66%) and myalgia (48%). The most meaningful laboratory results were: marked leucopenia (72%), thrombocytopenia (70-89%), hyponatremia (41%) and increased hepatic enzymes ALAT (41%), ASAT (45%) and LDH (62%). Dengue virus infection was diagnosed by means of a matching clinico epidemiological history and positivity of specific serology and/or virus isolation. Hemorrhagic phenomena appeared in 10 of the 71 patients (14%), out of which one was diagnosed with DHF according to WHO criteria. All patients recovered fully. CONCLUSION: Pretravel advice should be given to all travellers to dengue-endemic areas. DF must be included in the differential diagnosis of patients returning febrile from tropical areas. PMID- 14739028 TI - Trypanosoma cruzi transmission in a captive primate unit, Rio de Janeiro, Brazil. AB - A breeding in captivity program of neotropical primates for subsequent reintroduction in nature is in progress at the Primatology Center of Rio de Janeiro (CPRJ). Almost 200 animals of 20 species that include both wild captured animals and specimens born in captivity are maintained in CPRJ. Here, we examined 198 primates of CPRJ for infection with the protozoan parasite Trypanosoma cruzi. The animals included 18 species of eight genera. We also performed an "ad lib" search for triatomines that could be incriminated as putative transmitters of the protozoan in this scenario. Anti-T. cruzi antibodies were observed (by indirect immunofluorescence assay-IFA) in 40 monkeys (26.5%). Four Panstrongylus megistus were collected in the monkey's food storage room near the cages and in human dwellings in the proximity to CPRJ. T. cruzi were isolated from nine primates of two genera (Leontopithecus and Saguinus) and from two individuals of the vector P. megistus. The transmission inside the cages could be attested by the isolation of the T. cruzi from primates born in captivity. Multi-locus enzyme electrophoresis (MLEE) demonstrated that the two isolates from Saguinus bicolor bicolor displayed a zymodeme 1 profile in four out of five tested enzymes, while all isolates derived from Leontopithecus showed zymodeme 2 for four out of the five tested enzymes. Mini-exon gene analysis genotyped all isolates as T. cruzi II, which is associated with human disease in Brazil. A wild primate unit such as CPRJ, located inside the forest and near to human dwellings and with T. cruzi II infected animals, deserves a careful surveillance in order to prevent expansion of the infection. PMID- 14739029 TI - Haptoglobin phenotypes and malaria infection in pregnant women at delivery in western Cameroon. AB - Plasmodium falciparum-infected erythrocytes have been reported to sequester in the placenta by adhering to chondroitin 4-sulfate during pregnancy. Earlier studies have highlighted higher susceptibility of primigravidae to P. falciparum compared to multigravidae living within the same endemic areas. The haptoglobin phenotype (Hp1-1) has been associated with susceptibility to severe P. falciparum malaria and the presence of Hp in human endometrium has been reported. The possible role of different Hp phenotypes in susceptibility to or protection from placental infection by P. falciparum in both primigravid and multigravid women at delivery in western Cameroon was investigated in this study. Only the three major haptoglobin phenotypes; Hp1-1, Hp2-1 and Hp2-2, were found in the study population with the Hp1-1 phenotype being the predominant (53%). There was no significant difference in the distribution of the three Hp phenotypes between the two gravidity groups. Women carrying the Hp1-1 phenotype had higher parasite prevalences in both peripheral blood (21.6% against 9.1%) and placentas (42% against 16.7%) when compared to those with the Hp2-2 phenotype. The difference in the parasite density between women carrying the Hp1-1 and Hp2-2 phenotypes was statistically significant for placental infection (P=0.001) but not for maternal peripheral blood infection. Placental parasitaemias without peripheral blood parasitaemias were detected in 42.6% of all the P. falciparum positive women while 27.7% of the women had peripheral blood parasitaemias in the absence of placental infection and 29.8% of the women had both placental and peripheral blood parasitaemias. A statistically significant difference was observed between the primigravidae and multigravidae in the parasite density in placental biopsies (P=0.02) but not for maternal peripheral blood parasitaemia. Our data suggest that the Hp1-1 phenotype may play a role in susceptibility to placental infection by P. falciparum during pregnancy. PMID- 14739030 TI - Temperature preference in Rhodnius prolixus, effects and possible consequences. AB - The present work examines the thermal preference of adult Rhodnius prolixus along a temperature gradient. The mean preferred temperature differed slightly between sexes: 25.0 degrees C for males versus 25.4 degrees C for females. This preference was not constant, but varied daily by about 0.2 degrees C for both sexes, and reached its highest value at the onset of the dark phase and was lowest during the light phase. A change in the preferred temperature with the level of starvation was also observed (about 1 degrees C lower after 20 days of starvation). Changes in environmental temperature strongly affected the rate of weight loss for both sexes. When insects were maintained for 20 days in a chamber at 32 degrees C, they lost significantly more weight than when kept at 24 degrees C; both water loss and nutrient conversion processes are involved. This increase in weight loss rate with increasing temperature would cause a higher biting rate and consequently higher probability of Chagas' disease transmission. Females oviposit across a range of temperatures from 22 to 33 degrees C with a peak at 25 26 degrees C. These results are compared with patterns of thermopreference in other species of triatomine, as related to differences in their distribution and tolerance to starvation. PMID- 14739031 TI - Influence of the inoculation route in BALB/c mice infected by Leishmania infantum. AB - Mice of the BALB/c strain are frequently used, due to their high susceptibility to Leishmania infection. Most of the studies in visceral leishmaniasis use the endovenous or the intraperitoneal routes to inoculate the parasites. In this study, the development of experimental visceral leishmaniasis infection was evaluated in BALB/c mice inoculated with Leishmania infantum parasites by endovenous (EV group) or intraperitoneal (IP group) routes. The results shows that both inoculation routes were able to produce progressive infection in mice. However, a higher dispersion of the values of parasite density was detected among the animals of the EV group than the IP group. Our results indicate that the intraperitoneal inoculation results in a higher homogeneity of infections, so we suggest that this route should be preferentially used in experimental infections in the mice model, particularly when pools of samples are required for immune cellular studies. PMID- 14739032 TI - Environmental tobacco smoke exposure in women with lung cancer. AB - BACKGROUND: Investigations on environmental tobacco smoke (ETS) exposure that include source intensity, childhood exposure, and association with histologic subtypes among never smoking lung cancer cases are limited. We report the patterns of ETS exposure history in a clinical cohort of women with newly diagnosed lung cancer. METHODS: From 1997 to 2001, 810 women with lung cancer were interviewed to obtain data including the source, intensity, and duration of ETS exposure. In this descriptive study, relationships between smoking history, ETS exposure, and lung cancer histologic subtypes were analyzed. RESULTS: Among the 810 patients, 773 (95.4%) reported personal smoking or ETS exposure including 170 of 207 (82%) never smokers. Among the never smokers with a history of ETS exposure, the mean years of exposure were 27 from a smoking spouse, 19 from parents, and 15 from co-workers. For each major subtype of lung cancer (adenocarcinoma, squamous cell, unclassified non-small cell lung cancer, small cell, or carcinoids) among never smokers, 75-100% of patients had ETS exposure. Trends for adenocarcinoma, squamous, and small cell carcinoma are statistically significant using the Cochran-Armitage Test for Trend (P<0.001) among never smokers without ETS exposure, never smokers with ETS exposure, former smokers, and current smokers. CONCLUSIONS: Over 95% of women with lung cancer in our study were exposed to tobacco smoke through a personal smoking history or ETS. The cumulative amount of tobacco smoke exposure may be significantly underestimated if only personal smoking history is considered. Our results add to the public health implications of exposure to tobacco smoke and highlight the importance of eliminating tobacco smoking in public and private settings. PMID- 14739033 TI - Autocrine activation of ErbB2/ErbB3 receptor complex by NRG-1 in non-small cell lung cancer cell lines. AB - Our prior studies identified co-expression of the human epidermal growth factor like receptors 2 (ErbB2) and 3 (ErbB3), as well as the growth factor neuregulin-1 (NRG-1) in normal lung epithelium and lung cancers. As ErbB2 and ErbB3 dimerize to produce a high affinity receptor for NRG-1, we postulated that an autocrine growth loop was present in transformed and non-transformed pulmonary epithelial cells. To test this hypothesis, we examined four cell lines derived from human non-small cell carcinomas for: (1) ErbB2 and ErbB3 expression and endogenous activation; (2) NRG-1 expression and secretion/shedding; and (3) the effect of receptor blockade on autocrine receptor activation. Our studies found that ErbB2 and ErbB3 were expressed by each of these cell lines. In addition, the NRG-1 gene was also expressed with both major isoforms of NRG-1 (NRG-1alpha and NRG-1beta) found intracellularly. Only the NRG-1alpha isoform, however, was found secreted/shed into the culture medium. The secreted/shed NRG-1alpha was capable of activating the ErbB2/ErbB3 receptor complex expressed on the breast adenocarcinoma cell line MCF-7. Basal ErbB2 phosphorylation was identified in all lung cancer cell lines and was inhibited with an antibody that blocked the NRG-1 binding site on ErbB3. Taken together, these data show that secreted NRG-1alpha can activate the ErbB2/ErbB3 heterodimer in an autocrine fashion. The identification of a NRG-1alpha/ErbB2/ErbB3 autocrine loop raises the possibility that interruption of this loop may have therapeutic potential in lung cancer. PMID- 14739034 TI - Expression of thymidylate synthase is correlated with proliferative activity in non-small cell lung cancer (NSCLC). AB - Many experimental studies have revealed that enhanced thymidylate synthase (TS) expression is significantly correlated with higher proliferative activity of tumor cells, which may account for a poor prognosis of high-TS patients. However, only a few clinical studies have focused on the correlation between TS status and cell proliferation. Therefore, we assessed the correlation between TS expression and proliferative index (PI) as a marker of cell proliferation or p53 status in a total of 109 patients with completely resected pathologic stage I, non-small cell lung cancer (NSCLC). PI was defined as the percentage of tumor cells with positive staining against proliferative cell nuclear antigen (PCNA). The mean PIs of TS-high and TS-low tumors were 48.2% and 34.4% respectively, showing a significantly higher proliferative activity of TS-high tumor (P=0.020); when stratified according to histological type, the difference was significant in adenocarcinoma (P=0.038), but not in squamous cell carcinoma. There was no significant correlation between TS expression and p53 status. In conclusion, tumor cells with higher TS expression have higher proliferative activity in NSCLC, especially in adenocarcinoma. PMID- 14739035 TI - Survival and characteristics of lymph node involvement in patients with N1 non small cell lung cancer. AB - N1 non-small cell lung cancer (NSCLC) encompasses a heterogeneous subgroup with differential lymph node involvement. Among 738 patients with NSCLC who underwent surgical resection, including 579 patients (78.5%) with systematic hilar and mediastinal lymph nodal dissection, from 1992 to 2001, 82 patients were pathologically defined as having N1 disease. We retrospectively analyzed the factors influencing survival, including the characteristics of lymph node involvement; the location of involved stations, the number of involved stations, the number of involved nodes, and the status of nodal involvement (microscopic N1, nodal involvement first defined by postoperative histological examination; or macroscopic N1, nodal involvement obviously recognized by preoperative examinations or surgical explorations). The overall 5-year survival rate of the 82 patients with N1 disease was 50.9%. No significant differences in the overall survival were found with regard to gender, age, histologic type, type of resection, or adjuvant therapies. Pathologic T status significantly influenced the overall survival (T1 versus T2 disease, P=0.008). According to the characteristics of lymph node involvement, the prognosis of patients with multiple-node N1 involvement was significantly poorer than that of those with single-node N1 involvement (5-year survival: 29.6% versus 61.5%, p=0.003). The prognosis of patients with macroscopic N1 disease was significantly poorer than that of those with microscopic N1 disease (5-year survival: 43.0% versus 65.0%, P=0.046). By comparison with the survival of patients who underwent surgical resection during the same period for pathologic N0 (pN0) and pathologic N2 (pN2) diseases, no survival differences were observed between microscopic N1/single node N1 and pN0, or between multiple-node N1 and pN2 diseases. In patients with pathologic N1 disease, microscopic N1 and single-node N1 diseases may be an early stage, whereas multiple-node N1 disease behaves like an advanced stage. PMID- 14739036 TI - Impaired diffusion capacity predicts for decreased treatment tolerance and survival in limited stage small cell lung cancer patients treated with concurrent chemoradiation. AB - PURPOSE: To determine if stratification of limited stage small cell lung cancer (LSCLC) patients by pre-treatment pulmonary function test (PFT) prognostic indicators predicts for treatment-related toxicity risks and survival following concurrent chemoradiation. MATERIALS AND METHODS: From 1989 to 1999, 215 LSCLC patients received six cycles of alternating cyclophosphamide/doxorubicin/vincristine and etoposide/cisplatin (EP). Thoracic radiation (RT) was initiated only with EP and at cycle 2 or 3. RT dose was: 40 Gy/15 fractions/3 weeks or 50 Gy/25 fractions/5 weeks. RT fields encompassed gross and suspected microscopic disease with a 2 cm margin. Pre-treatment PFT values analyzed included forced expiratory volume in 1s (FEV1) (in liter and as % predicted) and diffusion capacity for carbon monoxide (DLCO) (as % predicted). The "marker" for toxicity during concurrent chemoradiation was the duration of any RT breaks initiated for severe hematologic or locoregional symptomatology. Patient outcomes were analyzed for associations between recognized PFT cut-offs (FEV1 <2l, > or =2l; FEV1 <60%, > or =60% predicted; DLCO <60%, > or =60% predicted), toxicity rates, and survival. RESULTS: For the whole study cohort, median, 2- and 5-year overall survivals were: 14.7 months, 22.7 and 7.2%, respectively. Fifty-six patients (26%) required treatment breaks due to toxicity. FEV1 and DLCO results were available for 96 (45%) and 86 (40%) patients, respectively. Two thirds of FEV1s measured were <2l. On statistical analysis, the incidence of toxicity-related interruptions was significant for DLCO<60% (P=0.043), suggestive for FEV1<2l (P=0.1) and non-significant for FEV1<60%. Patients with simultaneous DLCO<60% and FEV1<2l showed a trend toward increase toxicity risk (P=0.1). For selected PFT measures, median overall survivals were: 12.7 months versus 14.8 months for DLCO<60% versus > or =60%; 13.4 months versus 17.7 months for FEV1<2l versus > or =2l; 15.4 months versus 19.9 months for DLCO<60% + FEV1<2l versus DLCO> or =60% + FEV1> or =2l. Although absolute differences favored all patients with PFT values above the prognostic cut-offs, differences were not statistically significant on this analysis. Patients with both a treatment break and a DLCO<60% had the poorest median survival of all patient subsets, at 11.4 months (P=0.09). CONCLUSIONS: Impaired DLCO (i.e. <60%) is a novel predictor of increased treatment-related toxicity leading to interruptions. The present study suggests a probable role for DLCO and FEV1 (in l) as prognostic factors for predicting survival but larger patient samples are required for confirmation. Patients with impaired DLCOs experiencing treatment interruptions have the poorest survival. Assessment of pre-treatment PFTs contributes to determining optimal management strategies for LSCLC patients receiving definitive chemoradiation. PMID- 14739037 TI - A randomized trial comparing adjuvant chemotherapy versus surgery alone for completely resected pN2 non-small cell lung cancer (JCOG9304). AB - The purpose of this study was to evaluate the efficacy of adjuvant chemotherapy with three courses of cisplatin and vindesine, in comparison to observation only, for N2 non-small cell lung cancer that had been completely resected. Patients with pathologically demonstrated mediastinal lymph node metastasis (N2), who had undergone complete resection, were randomized to observation or adjuvant chemotherapy (cisplatin 80 mg/m2 on day 1; vindesine 3 mg/m2 on days 1 and 8: x3 courses). Cycles started within 6 weeks after complete resection and were repeated every 4 weeks. This trial was terminated before accumulation of the planned numbers for registration because of a slow accrual rate. A total of 119 patients were randomized (59 patients in the adjuvant arm and 60 with surgery alone). The median survival was 36 months for both groups. Postoperative cisplatin with vindesine chemotherapy was not shown to be efficacious in cases of completely resected N2 non-small cell lung cancer in this setting of timing, dose and agents studied. PMID- 14739038 TI - Quality of life in lung cancer: does disclosure of the diagnosis have an impact? AB - BACKGROUND: Quality of life (QOL) is an important component of evaluation in oncology. Usually, QOL is used in phase III studies to compare two treatments. The aim of this trial was to evaluate the impact of the disclosure of the diagnosis of cancer on QOL by using the European Organisation for Research and Treatment of Cancer core Quality of Life Questionnaire (EORTC QLQ)-C30 questionnaire and the supplemental lung cancer-specific module QLQ-LC13. PATIENTS AND METHODS: Patients hospitalised for exploration of an abnormal chest X-ray, with no previous history of cancer, a performance status < or =2, and able to fulfil the questionnaire were eligible. The patients answered the questionnaire two times: before (Q1) and after (Q2) the disclosure of the diagnosis. RESULTS: Seventy patients answered at Q1 and Q2. After the disclosure, some scores deteriorated: arm pain (P=0.009), physical functioning (P=0.01), role functioning (P=0.008), emotional functioning (P=0.0001) and social functioning (P=0.012), whereas the patients' own assessment of global QOL (item global QOL in functioning scales) did not show the same evolution. CONCLUSION: Disclosure of the diagnosis had an impact on social and emotional QOL. Patients with lung cancer need psychological support at the beginning of their disease. PMID- 14739039 TI - Quality of life assessment of second-line docetaxel versus best supportive care in patients with non-small-cell lung cancer previously treated with platinum based chemotherapy: results of a prospective, randomized phase III trial. AB - PURPOSE AND METHODS: Docetaxel second-line chemotherapy for non-small-cell lung cancer (NSCLC) has previously been shown to improve survival significantly compared with best supportive care (BSC). This multicenter phase III trial prospectively investigated quality of life (QOL) in NSCLC patients treated with either second-line docetaxel or BSC. Patients with stage IIIB/IV NSCLC, performance status < or =2, and adequate biochemistry and hematology were eligible if they had received > or =1 platinum-based chemotherapy regimens. Patients were randomized to docetaxel 100 mg/m2 (n=49) or, after protocol amendment, to docetaxel 75 mg/m2 (n=55), or to BSC (n=100), with Lung Cancer Symptom Scale (LCSS) and/or QLQ-C30 (with LC13 module) assessment every 3 weeks. RESULTS: Longitudinal analysis including all available assessments over time showed statistically significant differences in patient-rated pain scores in favor of docetaxel overall (P=0.005) or docetaxel 100 mg/m2 (P=0.003) compared to BSC. Trends in favor of docetaxel were noted on observer-rated scales for fatigue and pain for all docetaxel patients, and for total LCSS score, appetite and fatigue with docetaxel 100 mg/m2. Changes from baseline to the last available assessment (end point) showed significantly (P<0.05) less deterioration in LCSS pain score in patients with docetaxel (75 and 100 mg/m2 combined) than with BSC. An improved mean pain score with docetaxel 100 mg/m2 was contrasted with a deterioration in mean pain score with BSC (P<0.01). There was also significantly less deterioration in the global QOL today score with docetaxel 100 mg/m2 (P<0.01). Similar trends were recorded with QLQ-C30 assessments. CONCLUSIONS: Second-line docetaxel therapy for advanced NSCLC significantly improves survival with a trend towards less deterioration in QOL compared with BSC. PMID- 14739040 TI - Phase II study of bi-weekly docetaxel and carboplatin with concurrent thoracic radiation therapy followed by consolidation chemotherapy with docetaxel plus carboplatin for stage III unresectable non-small cell lung cancer. AB - OBJECTIVE: Docetaxel and carboplatin (DC) have demonstrated activity as radiation sensitizers in pre-clinical studies. The aim of this phase II study was to evaluate the efficacy and toxicity of DC with concurrent thoracic radiation therapy (TRT) followed by consolidation chemotherapy with DC for stage III unresectable non-small cell lung cancer (NSCLC). PATIENTS AND METHODS: Thirty three previously untreated patients with inoperable, locally advanced (LA) NSCLC received docetaxel 30 mg/m2 over 1 h and carboplatin at an AUC of 3 every 2 weeks for six courses--four courses during concurrent chemoradiotherapy and two courses following completion of radiotherapy. Concurrent TRT was performed in 2-Gy daily fractions to a total dose of 60 Gy. RESULTS: Among 32 evaluable patients, the overall response rate was 91%, with two complete responses (CR) and 27 partial responses (PR). Median survival time by intention-to-treat analysis was 27 months, with survival rates of 76% at 1 year and 61% at 2 years. Serious side effects were generally limited to grade 3 neutropenia in 6%, grades 3 and 4 pulmonary toxicity in 6 and 3%, respectively, and grade 3 esophagitis in 3% of patients. CONCLUSIONS: DC with concurrent TRT followed by consolidation chemotherapy was highly active with manageable toxicity in patients with stage III unresectable NSCLC. PMID- 14739041 TI - Phase I study of escalating doses of oxaliplatin in combination with fixed dose gemcitabine in patients with non-small cell lung cancer. AB - PURPOSE: Oxaliplatin (OHP) is a new platinum antineoplastic, while gemcitabine (GEM) is one of the most active drugs against non-small cell carcinoma (NSCLC). The OHP/GEM combination is interesting because the drugs have different mechanisms of action and toxicity profiles. The primary endpoint of this study was to determine the maximum tolerated dose (MTD) and dose-limiting toxicity (DLT) of OHP/GEM combination in non-small cell carcinoma of the lung. METHODS: Patients with relapsed NSCLC were treated with fixed dose i.v. GEM (1250 mg/m2) on days 1 and 8; followed on day 1 by i.v. OHP over 3 h, starting from 70 mg/m2 with 20 mg/m2 increments, up to 130 mg/m2. We enrolled 19 patients with eastern cooperative oncology group (ECOG) status 0/1=13/6; male/female=13/6. All had received first-line and four second-line chemotherapy. RESULTS: Four patients dropped out. At dose level 2, one patient died of pulmonary embolism; at level 3, two patients died of disease progression. One patient at level 3, refused to continue treatment after allergic reaction (high fever episode) during infusion of third cycle. Fifteen patients were evaluable for toxicity and response. According to a priori statistical considerations, three patients in each of the first three treatment levels and six in the last level were evaluable. No G3-4 toxicity was observed at levels 1 and 2. G3 neutropenia and anemia occurred in 8% of cycles at level 3. Six patients entered level 4 (OHP 130 mg/m2) with 22 courses delivered: G3-4 neutropenia occurred in 9%, G1-2 thrombocytopenia in 18%; other toxicities were mainly limited to G1-2 flu-like syndrome in about one third of patients and G1-2 nausea and vomiting in 5% of courses. There was no myelo-DLT at the highest dose level. There was no neurotoxicity at any level. Treatment was delayed in 12% and dose reduced in 26% of courses. There were 2/15 PR. CONCLUSIONS: MTD was not reached. OHP and GEM can probably be administered safely at 130 and 1250 mg/m2, respectively, as first-line therapy. The schedule is being used in a phase II trial. PMID- 14739042 TI - Treatment of malignant superior vena cava syndrome by endovascular stent insertion. Experience on 52 patients with lung cancer. AB - BACKGROUND: Superior vena cava syndrome (SVCS) is a frequent presentation of malignancies involving the mediastinum and can seriously compromise treatment options and prognosis. Stenting of superior vena cava is a well-known but not so commonly used technique to alleviate this syndrome. PATIENTS AND METHODS: Between August 1993 and December 2000 we performed 52 stenting procedures in patients affected by non-small cell lung cancer (NSCLC). RESULTS: Phlebographic resolution of the obstruction was achieved in 100% of cases with symptomatic and subjective improvement in more than 80%. One major complication was observed due to bleeding during anticoagulation. Re-obstruction of the stent occurred in only 17% of the cases, the majority due to disease progression. Improvement of the syndrome allowed hydration necessary for full dose platinum treatment when indicated in patients affected by lung cancer. CONCLUSIONS: Stenting of the superior vena cava syndrome is a safe and effective procedure achieving a rapid alleviation of symptoms in almost all patients, and allowing for full dose treatment in lung cancer patients. This procedure could change the traditional poorer prognosis attributed to non-small cell lung cancer patients presenting with this syndrome. PMID- 14739043 TI - Detection of p53 gene mutations in exhaled breath condensate of non-small cell lung cancer patients. AB - Early diagnosis of lung carcinoma is greatly desired. A potential source of early information regarding the process of cancerisation in the airways is exhaled breath condensate (EBC). The direct approach to detecting cancerisation is examining DNA from the area of chronic damage, i.e. airways and lung parenchyma. We therefore investigated DNA in EBC of patients with NSCLC and healthy volunteers. Human DNA was amplified by PCR in exhaled breath condensate and used to detect p53 mutations. A PCR of the beta-actin gene fragment was used to detect human DNA in each of the EBC samples. In 65.7% of the samples, the beta-actin gene was found. Extracted DNA as well as native EBC were equally suited as starting material for amplification. Mutations of the p53 gene were investigated in all EBC samples of NSCLC patients. p53 exons 5-8 were amplified using nested PCR and subsequently sequenced. Mutations were found in four of the patients (n=11; 36.4%) while no mutation was found in volunteers (n=10). Mutations detected in EBC were also compared with those of corresponding tumor tissue. Different point mutations in EBC and tumor tissue were revealed in all cases. Our findings demonstrate that exhaled breath condensate may be used for analysis of somatic gene mutations in an area of direct tobacco-related DNA damage. PMID- 14739044 TI - Practice guideline for the role of combination chemotherapy in the initial management of limited-stage small-cell lung cancer. AB - An evidence-based practice guideline was developed to identify the optimal combination chemotherapy regimen, schedule of administration, and duration of therapy for the first-line treatment of adults with limited-stage small-cell lung cancer. The guideline is based on a systematic search and review of literature published between 1985 and December 2002. Three reviewers selected studies for inclusion in the guideline according to pre-defined criteria. Fifty randomized controlled trials, five in abstract form, were included in the review, and feedback on a draft version of the guideline was obtained from medical oncologists in the province of Ontario, Canada. The most commonly used regimens in clinical trials are cyclophosphamide-doxorubicin(Adriamycin)-vincristine, and etoposide-cisplatin. No combination chemotherapeutic regimen has been conclusively shown to be superior to either of these regimens. Most studies comparing chemoradiation regimens used sequential rather than concurrent thoracic radiotherapy. When treating for cure with chemoradiation, there is evidence from one randomized controlled trial to support the use of etoposide-cisplatin over an anthracycline-containing regimen. There is conflicting evidence concerning a survival advantage for a regimen that alternates cyclophosphamide-doxorubicin vincristine with etoposide-cisplatin compared with either regimen alone. If bolus etoposide-cisplatin is the treatment of choice, evidence from one randomized trial suggests that the optimal sequence of administration is cisplatin followed by etoposide. The use of maintenance chemotherapy is not indicated. There is insufficient evidence to support the routine use of dose-intensive regimens outside a clinical trial, to determine the optimal duration of chemotherapy, or to support the routine substitution of carboplatin for cisplatin in combination chemotherapy regimens in this patient population. RECOMMENDATIONS: Etoposide cisplatin is the preferred chemotherapy regimen for patients with limited-stage small-cell lung cancer when concurrent thoracic radiotherapy is used. It is reasonable to offer the alternation of etoposide-cisplatin with cyclophosphamide doxorubicin-vincristine, provided the administration of radiotherapy concurrent with an anthracycline is avoided. PMID- 14739045 TI - Two-dimensional radiotherapy and docetaxel in treatment of stage III non-small cell lung carcinoma: no good survival due to radiation pneumonitis. PMID- 14739047 TI - Adipose tissue compartments and insulin resistance in overweight-obese Caucasian men. AB - We examined the association between insulin resistance and adipose tissue compartments in overweight/obese men. Total intra-abdominal, intraperitoneal, retroperitoneal, total subcutaneous, anterior subcutaneous and posterior subcutaneous abdominal adipose tissue (PSAAT) masses (total intra-abdominal fat TIAATM, IPATM, RPATM, TSAATM, ASAATM and PSAATM, respectively) were quantified in 51 overweight/obese men using magnetic resonance imaging (MRI). Total adipose tissue mass (TATM) was also determined using bioelectrical impedance. Insulin resistance was estimated by homeostasis model assessment (HOMA) score. In univariate regression analysis, all fat compartments, with the exception of RPATM, were significantly and positively correlated with the HOMA score. The positive correlation between HOMA score and both IPATM and anterior subcutaneous adipose tissue mass was independent of obesity status. After adjusting total for TATM, only IPATM was significantly correlated with HOMA score (partial r=0.38, P<0.01). In stepwise regression, IPATM was the best predictor of HOMA score. In multivariate regression models including age, obesity status, non-esterified fatty acid (NEFAs) levels, triglycerides and energy expenditure, IPATM remained a significant positive predictor (P<0.05) of HOMA score. In overweight/obese men intraperitoneal AT is the fat compartment that best predicts the degree of insulin resistance. This association appears to be independent of age, total body fat mass and obesity status. PMID- 14739048 TI - Pancreatic beta-cell defects in women at risk of type 2 diabetes. AB - We evaluated insulin release and insulin sensitivity in women with basal and/or postprandial hyperglycemia but normal oral glucose tolerance test (OGTT) in previous pregnancy (GHG). These women were individually matched with females without previous hyperglycemia (NGT). Both groups consisted of normal glucose tolerant women at the time of this study. They underwent OGTT (75 g; n=32 pairs) and hyperglycemic clamp experiments (10 mmoll(-1); n=27 pairs) with plasma glucose, insulin, and C-peptide measurements and calculation of insulinogenic index, first- and second-phase insulin release, and insulin sensitivity index (ISI). The GHG group showed higher glycosylated hemoglobin levels (6.2+/-0.6% versus 5.8+/-0.8%; P<0.05); lower insulinogenic index at 30 min (134.03+/-62.69 pmol mmol(-1) versus 181.59+/-70.26 pmol mmoll(-1); P<0.05) and diminished C peptide response in relation to glucose (4.05+/-0.36 nmol mmol(-1) versus 4.23+/ 0.36 nmol mmol(-1); P<0.05) at OGTT. Both groups did not show difference in insulin secretion and ISI by hyperglycemic clamp technique. We concluded that in up to 12 years from index pregnancy, women with previous GHG, presenting normal glucose tolerance and well-matched with their controls, showed beta-cell dysfunction without change in ISI. As women with previous GHG are at risk of type 2 diabetes, beta-cell dysfunction may be its primary defect. PMID- 14739049 TI - Tissue characterization identifies subjects with high risk of cardiovascular diseases. AB - OBJECTIVE: This study was conducted to examine whether tissue characterization of the carotid artery wall by determining integrated backscatter (IBS) can identify subjects with a recent history of acute coronary syndrome (ACS) or atherothrombotic infarction (ATI). RESEARCH DESIGN AND METHODS: The maximum thickness (Max-IMT) and IBS value (corrected-IBS) of the carotid artery intima media complex were measured ultrasonographically for 132 type 2 diabetic subjects (62.9+/-8.5 (+/-S.D.) years old, 87 men and 45 women) with or without cardiovascular diseases. RESULTS: The diabetic patients with recent ACS or ATI had a significantly lower corrected-IBS and higher Max-IMT than those with Max IMT > or = 1.1mm but without cardiovascular diseases. The product of Max-IMT and corrected-IBS (IMT-IBS product) of the patients with recent ACS was significantly lower than that of the patients with chronic stable angina. The multiple logistic model showed that only the IMT-IBS product was associated with a recent history of ACS (odds ratio 0.94, P<0.05) and ATI (odds ratio 0.90, P<0.005). CONCLUSIONS: Diabetic patients who had a recent history of ACS or ATI showed a lower IBS value and higher IMT for their carotid artery. Both the size and tissue characteristics of the carotid plaque may identify subjects with a high risk of cardiovascular disease. PMID- 14739050 TI - Long-term effects of a reduced fat diet intervention on cardiovascular disease risk factors in individuals with glucose intolerance. AB - The long-term effects on cardiovascular disease risk factors of a reduced fat (RF), ad libitum diet were compared with usual diet (control, CD) in glucose intolerance individuals. Participants were 136 adults aged > or =40 years with 'glucose intolerance' (2h blood glucose 7-11.0 mmol/l) detected at a Diabetes Survey who completed at 1 year intervention study of reduced fat, ad libitum diet versus usual diet. They were re-assessed at 2, 3 and 5 years. Main outcome measures were blood pressure, serum concentrations of total cholesterol, HDL and LDL cholesterol, total cholesterol:HDL ratio, triglycerides and body weight. The reduced fat diet lowered total cholesterol (P<0.01), LDL cholesterol (P< or =0.05), total cholesterol:HDL ratio (P< or =0.05), body weight (P<0.01) and systolic blood pressure (P< or =0.05) initially and diastolic blood pressure (P<0.01) long-term. No significant changes occurred in HDL cholesterol or triglycerides. In the more compliant 50% of the intervention group, systolic and diastolic blood pressure levels and body weight were lower at 1, 2 and 3 years (P<0.05). It was concluded that a reduced fat ad libitum diet has short-term benefits for cholesterol, body weight and systolic blood pressure and long-term benefits for diastolic blood pressure without significantly effecting HDL cholesterol and triglycerides despite participants regaining their lost weight. PMID- 14739051 TI - Death Certification and New Zealand Health Information Service (NZHIS) statistics for diabetes mellitus: an under-recognised health problem. AB - The main aims were to determine how much diabetes is under-recorded on death certificates, the influence of demographic variables and to what extent New Zealand Health Information Service (NZHIS) coding is able to compensate for deficiencies. A retrospective review of 600 death certificates (during 1999) from the Mortality Review Database in Christchurch Hospitals was carried out. Clinical data, laboratory results, coroner's reports, hospital clinical coding and NZHIS coding were collated and analysed. One hundred and four cases (17%) had clinically documented diabetes, of which only 47 (45%) had diabetes recorded on either the death certificate or the coroner's report. Diabetes was recognised in NZHIS coding in 45 (43%) of cases and in 94 (90%) cases in hospital clinical coding. Diabetes was more likely to be recorded on the death certificate in males and in subjects treated with insulin (P<0.05). Of the 496 subjects without documented diabetes, 159 (32%) had a highest random plasma glucose >11.1 mmol/l. Documented diabetes is thus under-reported on more than 50% of death certificates and this is not compensated by NZHIS coding. In addition, there is likely to be a significant number of cases with undiagnosed diabetes. This may adversely affect the reliability of mortality statistics. PMID- 14739052 TI - Predicting factors associated with costs of diabetic patients in Taiwan. AB - This study examined healthcare costs for medication-using diabetic patients in Taiwan and predicted which factors were associated with costs. We analyzed claims data from the National Health Insurance (NHI) program in Taiwan from 1998 to 1999. The approach included estimates of costs attributable to diabetes, diabetes related complications, comorbidity incurred by diabetic patients. A multiple regression model was used to assess the contribution of patients' characteristics in 1998 on outpatient, inpatient, and total costs in 1998 and 1999. The prevalence of medication-using patients with diabetes was 2.6% in 1998 and 2.8% in 1999. Costs of healthcare were 13.3% of total costs of NHI in 1998 versus 13.0% in 1999. Health services delivered near the end of life consumed large portions of medical dollars. The three most prevalent clinical associations of diabetes were congestive heart failure, neuropathy, and ischemic heart disease. Adjusted for demographic and clinical characteristics in 1998, this model could explain 8.0, 9.3, and 12.5%, respectively, of the cost variation in outpatient, inpatient, and total costs in 1999. PMID- 14739053 TI - Treatment of type 2 diabetes with a combination regimen of repaglinide plus pioglitazone. AB - The efficacy and safety of combination therapy (repaglinide plus pioglitazone) was compared to repaglinide or pioglitazone in 24-week treatment of type 2 diabetes. This randomized, multicenter, open-label, parallel-group study enrolled 246 adults (age 24-85) who had shown inadequate response in previous sulfonylurea or metformin monotherapy (HbA(1c) > 7%). Prior therapy was withdrawn for 2 weeks, followed by randomization to repaglinide, pioglitazone, or repaglinide/pioglitazone. In the first 12 weeks of treatment, repaglinide doses were optimized, followed by 12 weeks of maintenance therapy. Pioglitazone dosage was fixed at 30 mg per day. Baseline HbA(1c) values were comparable (9.0% for repaglinide, 9.1% for pioglitazone, 9.3% for combination). Mean changes in HbA(1c) values at the end of treatment were -1.76% for repaglinide/pioglitazone, 0.18% for repaglinide, +0.32% for pioglitazone. Fasting plasma glucose reductions were -82 mg/dl for combination therapy, -34 mg/dl for repaglinide, -18 mg/dl for pioglitazone. Minor hypoglycemia occurred in 5% of patients for the combination, 8% for repaglinide, and 3% for pioglitazone. Weight gains for combination therapy were correlated to individual HbA(1c) reductions. In summary, for patients who had previously failed oral antidiabetic monotherapy, the combination repaglinide/pioglitazone had acceptable safety, with greater reductions of glycemic parameters than therapy using either agent alone. PMID- 14739054 TI - Relationship between serum adiponectin and leptin concentrations and body fat distribution. AB - The aim of this study was to investigate the relationship between adiponectin and leptin and body fat distribution. One hundred and ninety-seven women participated in this study. Subjects were grouped based on their visceral adipose tissue area (VAT). Body fat distribution was determined by computed tomography. The numbers in the subcutaneous fat dominant group (SFDG) and visceral fat dominant group (VFDG) were 79 and 118, respectively. The VFDG showed lower adiponectin levels than the SFDG (8.9+/-0.4 microg/ml versus 11.4+/-0.7 microg/ml, P=0.006), but leptin levels did not differ significantly between groups (18.8+/-1.1 ng/ml versus 17.7+/-1.8 ng/ml, P=0.111). Adiponectin levels were inversely correlated with fasting insulin, HOMA-IR, triglyceride, SBP and DBP, subcutaneous adipose tissue area (SAT) and VAT, and waist-to-hip ratio (WHR). Leptin levels were positively correlated with fasting glucose and insulin, HOMA-IR, triglyceride, SBP and DBP, VAT and SAT, and WHR (all values of P<0.05). VAT and HDL-cholesterol were independent variables of adiponectin concentrations (R(2)=0.207, P<0.0001), and SAT, fasting insulin, and HOMA-IR were independent variables of leptin concentrations (R(2)=0.498, P<0.0001) In conclusion, adiponectin and leptin concentrations, although associated with metabolic parameters, were more strongly influenced by VAT in the case of adiponectin, and by SAT in the case of leptin. PMID- 14739055 TI - Non-alcoholic steatohepatitis in patients cared in metabolic units. AB - The clinical significance of liver disease is frequently underestimated in patients with metabolic disorders. In patients followed up in a metabolic unit for diabetes, obesity or hyperlipidemia (n=147), we studied the prevalence and the severity of liver disease, and its relationship with the metabolic syndrome (MS). Cases cared for in a liver unit (n=179) were used as controls. Patients in the metabolic series were older and had a higher prevalence of coronary heart disease. Criteria for the metabolic syndrome were fulfilled in 64% and 22% of cases, respectively (P<0.0001). Liver biopsy was obtained in 44 and 66% of cases. Metabolic patients had a more severe steatosis score (P<0.0001), whereas the scores of fibrosis and necroinflammation were less severe (P=0.0059 and 0.0007, respectively). Histological criteria for non-alcoholic steatohepatitis (NASH) were present in 82% of metabolic cases and 68% cases in the liver series (P=0.057). Liver disease in patients routinely cared for in metabolic units is similar to that observed in patients cared for in liver units, and potentially may progress to terminal liver failure. Liver biopsy is recommended for diagnostic and prognostic purposes, as well as for testing treatment effects in controlled trials. PMID- 14739056 TI - Prediction of hormone sensitivity by DNA microarray. AB - Endocrine-therapy continues to be extensively developed for treatment of breast cancer, and accurate therapeutic prediction of this hormone-associated cancer is strongly desired. Moreover, the role of estrogen and its receptor on the estrogen dependent growth of breast cancer cells has not been clarified hitherto. Thus, to develop a new diagnostic tool for endocrine-therapy, and to address the molecular mechanism of estrogen-dependent breast carcinogenesis, we investigated the gene expression profile of estrogen-responsive genes in breast cancer using DNA microarray technique. We first comprehensively analyzed the profile of estrogen responsiveness among several estrogen receptor (ER)-positive cancer cell lines by a large-scale DNA microarray. Based on the obtained information, a total of 138 genes which showed high induction or repression of the expression by estrogen stimulation were selected and provided for custom microarray. The results of the custom microarray analysis were consistent with those of large-scale microarray analysis, and revealed that they were clearly categorized into early- or late response types. Further analysis of these genes may provide new clues in the elucidation of the estrogen-dependent growth mechanisms of cancer. Furthermore, the custom microarray analysis of ER-positive breast cancer tissues also showed similar but not identical profiles to those of cell lines, indicating the potential of this custom microarray to predict the response to endocrine-therapy in the breast cancer. Moreover, in order to discover the new predictive factors for endocrine therapy in breast cancer patients, several candidate genes were selected and their expressions in breast cancer tissues were analyzed by real time RT-PCR and by immunohistochemical technique. These studies could provide new clues for elucidation of the estrogen-dependent mechanisms of cancer and clinical benefit for patients. PMID- 14739057 TI - Novel molecular aspects of prostate carcinogenesis. AB - Prostate adenocarcinoma is one of the most frequently diagnosed forms of cancer in the male population of the Western world. The pivotal role of androgen and its receptor in this disease has been abundantly documented and indeed, chemical castration and treatment with antiandrogens are now standard therapies. However, relapse is often observed after 18-24 months, due to the remarkable ability of prostate tumour cells to adapt to low or undetectable androgen levels. Amplification and mutations of the androgen receptor (AR) gene have been described as well as alterations in cofactor expression and crosstalk with other signalling pathways. Another recent line of research focused on the re programming of gene expression taking place in prostate tumours. Global expression profiling of normal and cancerous prostate tissues led to the identification of tumour-distinctive patterns. Validation studies are currently underway to identify novel drug targets as well as diagnostic and outcome prediction markers. PMID- 14739058 TI - The use of cytogenetics in understanding ovarian cancer. AB - The future of cancer research is no longer limited to epidemiological data and clinical management, but rather encompasses a new dimension of understanding, that involves genetics of the tumors themselves. This has been exemplified most prominently in hematological tumors where alterations at the DNA level have been found to play key roles in the pathophysiology, diagnosis, monitoring and prognosis of these tumors. It has been shown over the last 20 years that recurrent chromosomal rearrangements are strongly associated with the activation of oncogenes, acquisition of drug resistance and loss of tumor suppressor gene function. Chromosomal alterations have also been shown to characterize many solid tumors, including epithelial ovarian cancer [Cancer Res. 62 (2002) 3466; Cancer 91 (2001) 534; Genes Chromosomes Cancer 25 (1999) 290]. Despite these findings, however, there are currently few examples of specific cytogenetic studies that have contributed to the clinical management of solid tumors such as ovarian cancer. The limiting factor to date is the resolution of available techniques. With time, as the technology improves, so will our ability to focus on specific findings that may be applicable to future clinical management. The intention of this report is to familiarize the reader with the evolution of cytogenetic and molecular cytogenetic techniques used in the study of ovarian cancer, the early formulations from these studies and their use in answering specific clinical questions such as association with pathologic subtype, the relevance of drug resistance, the impact of BRCA mutations, and finally to guide the reader into the future of this ever growing field. PMID- 14739059 TI - Serum tumor markers in the management of ovarian, endometrial and cervical cancer. AB - CA 125 is the most reliable serum marker for ovarian carcinoma. Whereas its role in the screening of the malignancy is controversial, serum CA 125 assay is very useful for both the differential diagnosis of ovarian masses, particularly in postmenopause, and the monitoring of the response to chemotherapy and follow-up of patients with histologically proven ovarian carcinoma. Tumor-associated antigens other than CA 125, such as CA 19.9, CA 15.3 and TAG.72, firstly identified in gastro-intestinal or breast malignancies, have been detected also in tissue and serum samples from patients with ovarian carcinoma. In particular CA19.9 offers the advantage of high sensitivity for mucinous histotype, which often fails to express CA 125. Serum CA 125 correlates with the clinical course of disease better than the other antigens, and in patients with positive CA 125 assay at diagnosis the concomitant evaluation of CA 19.9 or CA 72.4 or CA 15.3 does not offer any additional benefit for monitoring ovarian carcinoma. Conversely, the serial measurements of these other antigens may represent an interesting biochemical tool for the management of patients with negative CA 125 assay. Serum alphaFP and betaHCG are very useful in the preoperative evaluation and management of nondysgerminomatous ovarian germ cell tumors, whereas elevated serum inhibin levels can be detected in patients with granulosa cell tumors of the ovary. As for endometrial carcinoma, preoperative serum CA 125 levels correlate with stage, depth of myometrial invasion, histologic grade, cervical invasion, peritoneal cytology, lymph node status and clinical outcome. Moreover, serial CA 125 assay is a good indicator of disease activity and a useful biochemical tool for post-treatment surveillance of patients with endometrial carcinoma. SCC is the most reliable serum marker for squamous cell cervical carcinoma, and in patients with this malignancy pretreatment SCC levels are related to tumor stage, tumor size, depth of cervical invasion, lymph-vascular space involvement, lymph node status and clinical outcome. Serial SCC measurements parallel the response to radiotherapy and chemotherapy as well as the clinical course of disease after the completion of treatment. Serum CYFRA 21.1 seems to be less sensitive than serum SCC for squamous cell cervical carcinoma. Elevated CA 125 levels can be often detected in patients with cervical adenocarcinoma. The future for tumor marker research is represented by the emerging technologies of transcriptional profiling and proteomics. PMID- 14739060 TI - Neurodegenerative diseases and oxidative stress. AB - Oxidative stress is now recognized as accountable for redox regulation involving reactive oxygen species (ROS) and reactive nitrogen species (RNS). Its role is pivotal for the modulation of critical cellular functions, notably for neurons astrocytes and microglia, such as apoptosis program activation, and ion transport, calcium mobilization, involved in excitotoxicity. Excitotoxicity and apoptosis are the two main causes of neuronal death. The role of mitochondria in apoptosis is crucial. Multiple apoptotic pathways emanate from the mitochondria. The respiratory chain of mitochondria that by oxidative phosphorylation, is the fount of cellular energy, i.e. ATP synthesis, is responsible for most of ROS and notably the first produced, superoxide anion (O(2)(;-)). Mitochondrial dysfunction, i.e. cell energy impairment, apoptosis and overproduction of ROS, is a final common pathogenic mechanism in aging and in neurodegenerative disease such as Alzheimer's disease (AD), Parkinson's disease (PD) and amyotrophic lateral sclerosis (ALS). Nitric oxide (NO(;)), an RNS, which can be produced by three isoforms of NO-synthase in brain, plays a prominent role. The research on the genetics of inherited forms notably ALS, AD, PD, has improved our understanding of the pathobiology of the sporadic forms of neurodegenerative diseases or of aging of the brain. ROS and RNS, i.e. oxidative stress, are not the origin of neuronal death. The cascade of events that leads to neurons, death is complex. In addition to mitochondrial dysfunction (apoptosis), excitotoxicity, oxidative stress (inflammation), the mechanisms from gene to disease involve also protein misfolding leading to aggregates and proteasome dysfunction on ubiquinited material. PMID- 14739061 TI - Sulfur containing amino acids and human disease. AB - Sulfur containing amino acids contribute substantially to the maintenance and integrity of cellular systems by influencing cellular redox state and cellular capacity to detoxify toxic compounds, free radicals and reactive oxygen species. Methionine and cysteine are the two primary sulfur-containing amino acids in mammals. Methionine is an essential amino acid, obtained by dietary intake while cysteine is non-essential and a metabolite of methionine metabolism. Each of these amino acids contributes significantly to the cellular pool of organic sulfur and generally to sulfur homeostasis as well as playing a significant role in regulation of one carbon metabolism. Genetic defects in the enzymes regulating sulfur pools produce a variety of human pathologies, including homo- and cystinuria, homo- and cysteinemia, and neural tube defects. In addition, thiol imbalance has been associated with multiple disorders, including vascular disease, Alzheimer's, HIV and cancer. Possible treatments to restore the thiol balance are also discussed. PMID- 14739062 TI - Accuracy of fine-needle aspiration cytology and frozen-section examination in patients with thyroid cancer. AB - In patients with solitary thyroid nodules (TN) undergoing surgery both fine needle aspiration (FNA) and frozen-section examination (FSE) are currently performed, but their role is still controversial. We retrospectively analyzed a series of 606 patients with a nontoxic solitary thyroid nodule who underwent both FNAB and FSE prior to thyroidectomy. There were 118 (19.5%) men and 488 (80.5%) women, with a median age of 44 years (range 16-81 years). The results of both FNAB and FSE were compared against the final pathological examination. Definitive histology showed 500 (82.5%) benign nodules, including 239 (39.4%) follicular adenomas, and 106 (17.5%) carcinomas, of which 18 (17.0%) were follicular cancer. In differentiating between benign TN and thyroid tumors the sensitivity, specificity, and accuracy of FNAB were 93.6%, 98.9% and 95.9%, while in the detection of malignancy were 81.1%, 99.4% and 96.2% for FNAB, and 83.0%, 100% and 97.0% for FSE (P = NS), respectively. The combination of both techniques did not improve significantly (P = NS) the results. Both medullary and undifferentiated cancer, and 76 of 80 (95.0%) papillary carcinomas were correctly detected by FNAB. Three (0.5%) false-positive smears suggesting cancer were found. In conclusion, in patients with FNAB revealing thyroid cancer or a benign TN, the extent of thyroidectomy should be established by performing FSE. When FNAB suggests the presence of a follicular tumor, FSE may affect rarely the final intraoperative decision-making, and should be considered unnecessary. However, in patients with FNA revealing thyroid cancer, FSE is useful in confirming malignancy, and may avoid surgical overtreatments. PMID- 14739063 TI - Identification of retroviral conserved pol sequences in serum of mixed connective tissue disease and systemic sclerosis patients. AB - Human endogenous retroviruses (HERVs) expression has been suspected as a factor participating in the development of autoimmune diseases. Employing the dot blot hybridisation, we observed that 90.9% of mixed connective tissue disease (MCTD) (n = 22) patients exhibited the presence of human immunodeficiency virus type 1 (HIV-1) conserved pol sequence in DNA isolated from serum. The same investigation conducted in the group of systemic sclerosis (SSc) (n = 79) patients identified a conserved retroviral pol sequence in 34.6% of antinuclear antibodies (ANA) positive (n = 52) and 40.7% of ANA negative (n = 27) patients. However, 100% of anti-U1-ribonucleoprotein (RNP) positive SSc (n = 9) patients exhibited the presence of HIV-1 conserved pol sequence. Our findings suggest that the presence of retroviral sequence in serum of MCTD and SSc patients may correlate with development of autoimmune response directed against U1-70 kDa polypeptide antigen. PMID- 14739064 TI - Effects of L-fucose and fucose-rich oligo- and polysaccharides (FROP-s) on collagen biosynthesis by human skin fibroblasts. Modulation of the effect of retinol, ascorbate and alpha-tocopherol. AB - The action of L-fucose and a fucose-rich oligosaccharide (FROP-3) on skin explant cultures and fibroblast cell cultures, alone or together with three vitamins (A, C, E, often used in topical preparations) was studied, using as criteria collagen accumulation and collagen and non-collagen protein biosynthesis. In order to confirm the results observed, several variants of the chemical procedure were used. Collagen accumulation in cell cultures and (3)H-proline incorporation in proteins and collagen gave comparable results. L-Fucose produced a slight but significant inhibition of collagen accumulation and of incorporation of radioactive proline in cell associated collagen and also modified the ratio of excreted (soluble) to cell- or tissue-bound collagen and non-collagen proteins. The oligosaccharide (FROP-3) did not produce a comparable inhibition of collagen biosynthesis. Both L-fucose and FROP-3 modulated the action of the two above mentioned vitamins in most experimental conditions used. The combined action of the three vitamins (all-trans retinol, ascorbate, alpha-tocopherol) with L-fucose and even more so with FROP-3 can be considered as favourable for the modulation of the biosynthetic activity of fibroblasts. PMID- 14739070 TI - New developments in nutrition and diabetes: glyceroneogenesis comes of age. PMID- 14739071 TI - Glyceroneogenesis revisited. AB - Glyceroneogenesis is the synthesis of 3-glycerol phosphate by an abbreviated version of gluconeogenesis. The research that led to the discovery of glyceroneogenesis in white adipose tissue is presented. This pathway is active during fasting in white and brown adipose tissue and in the liver as part of the triglyceride/fatty acid cycle. Glyceroneogenesis is critical for the extensive recycling of free fatty acid (FFA) back to triglyceride that occurs in mammals, including humans, after lipolysis, when up to 65% of the fatty acids are re esterified back to triglyceride. The rate-limiting enzyme in this pathway is the cytosolic form of phosphoenolpyruvate carboxykinase (GTP) (4.1.1.32) (PEPCK-C). Transcription of this gene is induced in adipose tissue and liver during fasting. Ablation of expression of the gene for PEPCK-C in white adipose tissue of mice results in lipodsytrophy, while overexpression of the gene for this enzyme in adipose tissue causes obesity. The critical role of glyceroneogenesis in diabetes was suggested by experiments in which the gene for PEPCK-C is induced in white adipose tissue by rosiglitazone, a drug used to control diabetes in humans. This was accompanied by a marked decrease in FFA release from adipose tissue due to an induction in glyceroneogenesis in the tissue. Since the chronic release of FFA by adipose tissue is a critical factor in the development Type 2 diabetes, it is likely that rosiglitazone acts in part by stimulating transcription of the gene for PEPCK-C, thereby increasing rate of glyceroneogenesis and lowering the rate of FFA release from adipose tissue. PMID- 14739072 TI - Regulation of cytosolic phosphoenolpyruvate carboxykinase gene expression in adipocytes. AB - Cytosolic phosphoenolpyruvate carboxykinase (EC 4.1.1.32; PEPCK-C) catalyzes the critical regulated step in adipocyte glyceroneogenesis. Numerous studies have shown that hormones and nutrients regulate PEPCK-C at the transcriptional level. We identified two upstream cis-acting DNA elements, gAF1/PCK1 and PCK2, that control adipocyte specific transcription of the PEPCK-C gene (Pck1). Both elements are direct repeat hexanucleotides separated by 1 bp (DR1 elements; variations of the sequence AGGTCAnAGGTCA). PCK2 is located 1 kbp upstream and is the essential element of an adipocyte specific enhancer. It is a peroxisome proliferator activated receptor gamma response element (PPRE) and directs the activation of the PEPCK-C gene during adipogenesis. In addition, it is a thiazolidinedione response element in mature adipocytes. In contrast, gAF1/PCK1, centered 445 bp upstream, is a pleiotropic element that mediates tissue specific glucocorticoid action-repression in adipocytes and induction in hepatocytes. It is a negative response element for PPARgamma, RXRalpha, COUP-TFII, and several unidentified proteins in some cell types, and a positive element for COUP-TFI and HNF4 in other cells type. The purpose of this presentation is to review the discovery and characterization of these two elements in adipocytes and describe how our work has contributed to understanding the mechanisms that control adipocyte glyceroneogenesis. PMID- 14739073 TI - Regulation of glyceroneogenesis and phosphoenolpyruvate carboxykinase by fatty acids, retinoic acids and thiazolidinediones: potential relevance to type 2 diabetes. AB - Recent studies brought adipocyte glyceroneogenesis back to the fore as an important pathway in fatty acid homeostasis and underlined the key role played by cytosolic phosphoenolpyruvate carboxykinase (PEPCK-C) in this pathway. The present review analyses the mechanisms by which a series of hormones and nutrients affect PEPCK-C gene transcription and glyceroneogenesis and describes evidence for disregulation of this pathway in type 2 diabetes. PMID- 14739074 TI - Central role of the adipocyte in the insulin-sensitising and cardiovascular risk modifying actions of the thiazolidinediones. AB - Insulin resistance is a key metabolic defect in type 2 diabetes that is exacerbated by obesity, especially if the excess adiposity is located intra abdominally/centrally. Insulin resistance underpins many metabolic abnormalities collectively known as the insulin resistance syndrome-that accelerate the development of cardiovascular disease. Thiazolidinedione anti-diabetic agents improve glycaemic control by activating the nuclear receptor peroxisome proliferator activated receptor-gamma (PPARgamma). This receptor is highly expressed in adipose tissues. In insulin resistant fat depots, thiazolidinediones increase pre-adipocyte differentiation and oppose the actions of pro-inflammatory cytokines such as tumour necrosis factor-alpha. The metabolic consequences are enhanced insulin signalling, resulting in increased glucose uptake and lipid storage coupled with reduced release of free fatty acids (FFA) into the circulation. Metabolic effects of PPARgamma activation are depot specific-in people with type 2 diabetes central fat mass is reduced and subcutaneous depots are increased. Thiazolidinediones increase insulin sensitivity in liver and skeletal muscle as well as in fat, but they do not express high levels of PPARgamma, suggesting that improvement in insulin action is indirect. Reduced FFA availability from adipose tissues to liver and skeletal muscle is a pivotal component of the insulin-sensitising mechanism in these latter two tissues. Adipocytes secrete multiple proteins that may both regulate insulin signalling and impact on abnormalities of the insulin resistance syndrome--this may explain the link between central obesity and cardiovascular disease. Of these proteins, low plasma adiponectin is associated with insulin resistance and atherosclerosis- thiazolidinediones increase adipocyte adiponectin production. Like FFA, adiponectin is probably an important signalling molecule regulating insulin sensitivity in muscle and liver. Adipocyte production of plasminogen activator inhibitor-1 (PAI-1), an inhibitor of fibrinolysis, and angiotensin II secretion are partially corrected by PPARgamma activation. The favourable modification of adipocyte-derived cardiovascular risk factors by thiazolidinediones suggests that these agents may reduce cardiovascular disease as well as provide durable glycaemic control in type 2 diabetes. PMID- 14739075 TI - Immortalization of human preadipocytes. AB - Primary human preadipocytes in culture are characterized by a low proliferative capacity associated with a rapid decline of differentiation ability during subculturing; thereby limiting their use as cellular model. Cellular immortalization constitutes an interesting approach for establishing cell lines presenting an unlimited life span and a maintained differentiation capacity. Different procedures for developing immortalized human preadipocytes are discussed in this review. Transformation of human preadipocytes with the simian virus 40 large T-antigen (SV40 T-Ag) permitted the development of immortalized cells; however these cells could not maintain their capacity to differentiate into adipocytes. This limitation may be explained by the ability of SV40 T-Ag to inhibit transcriptional factors involved in the differentiation of preadipocyte. Reconstitution of the telomerase activity by stable expression of the hTERT (human telomerase catalytic subunit) gene was able to partially extend the lifespan of primary preadipocytes but not to promote cellular immortalization. However, a combined expression of hTERT and the E7 oncoprotein of human papillomavirus type 16, generated human preadipocytes with both an unlimited life span and a preserved adipogenic potential. This approach appears to be an effective method for establishing human preadipose cell lines for studying adipocyte differentiation and metabolism. PMID- 14739076 TI - Protein and lipid analysis of detergent-resistant membranes isolated from bovine kidney. AB - Detergent-resistant membranes (DRM) were prepared from bovine kidney cortex. The criterion used to test their purification was the increase in the activity of a GPI membrane-anchored protein, the alkaline phosphatase. Its association with specific proteins and lipids was tested. Two successive Triton X-100 treatments followed by purification on sucrose gradient at 4 degrees C were necessary to obtain DRM with a maximum of alkaline phosphatase activity and a typical protein pattern. A third Triton treatment did not alter this DRM composition. Among the enriched protein, we identified, by mass spectrometry, a microsomal dipeptidase, which was GPI membrane-anchored. Protein-kinase activities, mainly serine-kinase, were enriched during the DRM purification. Using the typical FTIR olefinic =C-H bands of the acyl chains, a global decrease in the unsaturation level of DRM lipids was observed as compared with total membranes. Three main phospholipids were identified in DRM. Their fatty acid compositions were determined by gas chromatography and compared with those of total membranes. The most enriched saturated fatty acid was palmitic acid (+44% for phosphatidylethanolamine, +52% for phosphatidylcholine and +49% for sphingomyelin), agreeing with a selection of specific phospholipids among the saturated ones during the DRM purification. PMID- 14739078 TI - Expression of phosphoenolpyruvate carboxykinase gene in human adipose tissue: induction by rosiglitazone and genetic analyses of the adipocyte-specific region of the promoter in type 2 diabetes. AB - Cytosolic phosphoenolpyruvate carboxykinase (PEPCK-C) is the key enzyme in glyceroneogenesis, an important metabolic pathway in adipocytes for reesterification of fatty acids during fasting. Dysregulation of glyceroneogenesis could play a role in the increase in plasma non-esterified fatty acids that accompanies type 2 diabetes. In rodent adipocyte transcription of the PEPCK-C gene is induced by thiazolidinediones (TZDs) through an element, named PCK2, in its promoter. PCK2 binds a peroxisome proliferator activated receptor gamma (PPARgamma), retinoid X receptor alpha (RXRalpha) heterodimer. We demonstrated that in cultured human subcutaneous adipose tissue explants, PEPCK-C specific activity and mRNA were induced by 1 microM of the TZD rosiglitazone, respectively, about twofold in 8 h and fivefold in 5 h. Using gel shift experiments, we show that this effect is likely to involve the human PCK2 (hPCK2) element, which binds a protein complex that contains PPARgamma and RXRalpha. We analyzed hPCK2 (position -1031 to -1015 base pairs) and nearby sequences in the PEPCK-C promoter in 403 subjects with type 2 diabetes and 123 non-diabetic controls. The sequence of hPCK2 was not polymorphic, but we detected two C/T single nucleotide polymorphisms (SNPs), in complete linkage disequilibrium, at positions -1097 and -967 bp. Allele and genotype frequencies were not significantly different in patients and controls. However, our results suggest co dominant effects of C and T-alleles on fasting plasma glucose and glycosylated hemoglobin A1c levels in obese type 2 diabetic patients. PMID- 14739077 TI - Analysis of lipolysis in adipocytes using a fluorescent fatty acid derivative. AB - For facilitation of the experimental analysis of the mechanism and regulation of mobilization of fatty acids from adipose triacylglycerol (TAG) stores, which also represents important targets for pharmacological intervention with the pathogenesis of diabetes and obesity, we developed a convenient and reliable non radioactive cell-based assay. Isolated rat adipocytes are incubated with the fluorescent fatty acid derivative, 12-((7-nitrobenz-2-oxa-1,3-diazol-4 yl)amino)dodecanoic acid (NBD-FA), in the presence of insulin. The resulting NBD FA-labeled TAG is efficiently cleaved by hormone-sensitive lipase (HSL) in vitro. After removal of insulin and excess of free NBD-FA, lipolysis is initiated by addition of isoproterenol and/or adenosine deaminase. The amount of NBD-FA generated in total or released into the incubation medium in the presence of modulatory hormones or compounds is then monitored by thin layer chromatography and fluorescence imaging. Release of NBD-FA, glycerol and [3H]oleic acid from TAG follows similar kinetics and concentration dependence in response to various lipolytic and anti-lipolytic stimuli as well as inhibitors of HSL. Release of NBD FA from adipocytes correlates well to translocation of HSL from the cytosol to TAG droplets. In addition, we found that a cell-free system consisting of NBD-FA labeled TAG droplets with endogenous associated HSL closely reflects the lipolytic state of the adipocytes used for its preparation. In conclusion, release of NBD-FA from TAG in vivo and in vitro can be used as accurate index for (regulation of) lipolysis in primary and cultured adipocytes. PMID- 14739079 TI - 1964: The first model for the shape of a transfer RNA molecule. An account of an unpublished small-angle X-ray scattering study. AB - The shape of non-fractionated Escherichia coli transfer RNA molecules in solution was investigated using small-angle X-ray scattering during the years 1960-1962 at the Centre de Recherche sur les Macromolecules in Strasbourg. The innermost region of the scattering curve yielded the average molecular weight (Mr) and the radius of gyration (Rg) of the particles, whereas the experimental data at large angles could be approximated at best by the scattering curve of a kinked rod shaped molecule. The simplest model that was compatible with Mr, Rg, and the mass per unit length of the rod was a boomerang-shaped particle made of two double helical stems connected by a sharp kink. This model that eventually proved similar to the high-resolution L-shaped structure, was presented in my Ph.D. dissertation (J. Witz, Etude de la structure de quelques polynucleotides en solution par diffusion centrale des rayons X, Ph.D. dissertation, University of Strasbourg, France, 1964) but has never been published in detail. It is the purpose of this note to recall this story. PMID- 14739080 TI - Effects of education, vaccination and treatment on HIV transmission in homosexuals with genetic heterogeneity. AB - Genetic studies report the existence of a mutant allele Delta32 of CCR5 chemokine receptor gene at high allele frequencies (approximately 10%) in Caucasian populations. The presence of this allele is believed to provide partial or full resistance to HIV. In this study, we look at the impact of education, temporarily effective vaccines and therapies on the dynamics of HIV in homosexually active populations. In our model, it is assumed that some individuals possess one or two mutant alleles (like Delta32 of CCR5) that prevent the successful invasion or replication of HIV. Our model therefore differentiates by genetic and epidemiological status and naturally ignores the reproduction process. Furthermore, HIV infected individuals are classified as rapid, normal or slow progressors. In this complex setting, the basic reproductive number R0 is derived in various situations. The separate or combined effects of therapies, education, vaccines, and genetic resistance are analyzed. Our results support the conclusions of Hsu Schmitz that some integrated intervention strategies are far superior to those based on a single approach. However, treatment programs may have effects which counteract each other, as may genetic resistance. PMID- 14739081 TI - Comparative methods based on species mean values. AB - Comparative methods that use simple linear regression based on species mean values introduce three difficulties with respect to the standard regression model. First, species values may not be independent because they form part of a hierarchically structured phylogeny. Second, variation about the regression line includes two sources of error: 'biological error' due to deviations of the true species mean values from the regression line and sampling error associated with the estimation of these mean values [B. Riska, Am. Natural. 138 (1991) 283]. Third, sampling error in the independent variable results in an attenuated estimate of the regression slope. We consider estimation and hypothesis testing using two statistical models which explicitly justify the use of the species mean values, without the need to account for phylogenetic relationships. The first (random-effects) is based on an evolutionary model whereby species evolve to fill a bivariate normal niche space, and the second (fixed-effects) is concerned with describing a relationship among the particular species included in a study, where the only source of error is in the estimation of species mean values. We use a modification of the maximum-likelihood method to obtain an unbiased estimate of the regression slope. For three real datasets we find a close correspondence between this slope and that obtained by simply regressing the species mean values on each other. In the random effects model, the P-value also approximates that based on the regression of species mean values. In the fixed effects model, the P value is typically much lower. Simulated examples illustrate that the maximum likelihood approach is useful when the accuracy in estimating the species mean values is low, but the traditional method based on a regression of the species mean values may often be justified provided that the evolutionary model can be justified. PMID- 14739082 TI - A resistor interpretation of general anisotropic cardiac tissue. AB - This paper describes a spatial discretization scheme for partial differential equation systems that contain anisotropic diffusion. The discretization method uses unstructured finite volumes, or the boxes, that are formed as a secondary geometric structure from an underlying triangular mesh. We show how the discretization can be interpreted as a resistive circuit network, where each resistor is assigned at each edge of the triangular element. The resistor is computed as an anisotropy dependent geometric quantity of the local mesh structure. Finally, we show that under certain conditions, the discretization gives rise to negative resistors that can produce non-physical hyperpolarizations near depolarizing stimuli. We discuss how the proper choice of triangulation (anisotropic Delaunay triangulation) can ensure monotonicity (i.e. all resistors are positive). PMID- 14739083 TI - The asymptotic behavior of a chemostat model with the Beddington-DeAngelis functional response. AB - In this paper, the global asymptotic behavior of a chemostat model with Beddington-DeAngelis functional response is studied. The conditions for the global asymptotical stability of the model with time delays are obtained via monotone dynamical systems. Our results demonstrate that those time delays affect the competitive outcome of the organisms. PMID- 14739084 TI - A phase transition for a random cluster model on phylogenetic trees. AB - We investigate a simple model that generates random partitions of the leaf set of a tree. Of particular interest is the reconstruction question: what number k of independent samples (partitions) are required to correctly reconstruct the underlying tree (with high probability)? We demonstrate a phase transition for k as a function of the mutation rate, from logarithmic to polynomial dependence on the size of the tree. We also describe a simple polynomial-time tree reconstruction algorithm that applies in the logarithmic region. This model and the associated reconstruction questions are motivated by a Markov model for genomic evolution in molecular biology. PMID- 14739085 TI - A growth-controlled model of the shape of a sunflower head. AB - A mathematical model is presented which predicts the shape of a sunflower receptacle (or the compact receptacle of various other taxa) and the pattern of its floral parts (florets) from the time of their initiation to maturity. The model assumes that the expansion and curving of the receptacle surface is just sufficient to accommodate the development of the florets, thus minimizing the quantity of plant tissue involved. The model assumes a fixed angular separation (divergence) between successive florets, an S-shaped (sigmoidal) growth function followed by each florets, and a fixed time delay (period) between the initiation of successive florets. It is further assumes that the shape and relative position occupied by the florets on the receptacle surface are invariant in time. By this theory, the shape of the receptacle surface is fully determined once the mathematical form of the growth function is specified. Using the logistic growth function, the theory is tested against the measured shapes of plant receptacles from different taxa at various points in their development. The least-squares adjusted fits to the theory are, in most cases, very good indeed. PMID- 14739087 TI - Liver transplantation in adults with acute liver failure. PMID- 14739088 TI - The unexpected outcomes of medical research: serendipity and the microsomal ethanol oxidizing system. PMID- 14739089 TI - Lack of evidence for reovirus infection in tissues from patients with biliary atresia and congenital dilatation of the bile duct. AB - BACKGROUND/AIMS: To clarify the association between the reovirus infection of the hepatobiliary tree and the development of infantile obstructive cholangiopathy (IOC) including biliary atresia (BA) and congenital dilatation of the bile duct (CBD). METHODS: We designed reovirus common primers for nested RT-PCR based on the L3 gene segment. The spectrum and the sensitivity of common primers were evaluated with purified reoviral RNAs and reovirus mixed with stool samples. Then, nested RT-PCRs were performed with hepatobiliary and fecal samples obtained from patients with BA, CBD, and control diseases. Additionally, electron microscopy of stool samples was performed. RESULTS: The L3 common primers could amplify cDNAs synthesized from RNAs of three prototypes of reovirus, and detect as much as 5.0x10(3) plaque forming unit of serotype 3 Dearing strain in 100 mg of fecal samples. However, no amplification product was detected in 136 hepatobiliary tissues taken from 67 patients including 26 BAs and 28 CBDs, or in 65 fecal samples obtained from 15 patients including 10 BAs and 1 CBD. Additionally, viral particles were not found in any stool specimens by the electron microscope. CONCLUSIONS: These data do not suggest that reoviruses play a major role in the etiology of IOC or BA. PMID- 14739090 TI - The human organic anion transporting polypeptide 8 (SLCO1B3) gene is transcriptionally repressed by hepatocyte nuclear factor 3beta in hepatocellular carcinoma. AB - BACKGROUND/AIMS: The organic anion transporting polypeptides (OATPs) mediate the uptake of numerous amphipathic compounds into hepatocytes. Our aim was to study the expression and regulation of OATP8 (OATP1B3, SLC21A8/SLCO1B3) and OATP-C (OATP1B1, SLC21A6/SLCO1B1) in hepatocellular carcinomas (HCC). METHODS: RNA and protein levels in 13 paired HCC and adjacent non-tumor liver samples were quantified by real-time polymerase chain reaction or Western blot, respectively. The OATP8 and OATP-C gene promoters were characterized by luciferase reporter assays and electrophoretic mobility shift assays (EMSA). RESULTS: The expression of OATP8 was decreased in 60% of HCC compared to surrounding non-tumor liver tissue, on both the mRNA and protein levels. Expression of the liver-enriched transcription factor hepatocyte nuclear factor 3beta (HNF3beta) was increased in 70% of HCC and correlated inversely with OATP8 mRNA (r=-0.75, P<0.05) and protein. In contrast to OATP8, expression of OATP-C was not significantly decreased in HCC. In transfected Huh7 cells, OATP8 promoter activity was inhibited by 70% when HNF3beta was cotransfected. An HNF3beta binding site was located at nt -39/-23 by EMSA. The OATP-C promoter was not inhibited by HNF3beta. CONCLUSIONS: HNF3beta represses transcription of the OATP8 but not the OATP-C gene, providing a mechanism for reduced expression of OATP8 in HCC. PMID- 14739091 TI - Large-scale gene profiling of the liver in a mouse model of chronic, intragastric ethanol infusion. AB - BACKGROUND/AIMS: The mechanisms underlying alcohol-induced liver injury are not fully elucidated. An approach in this direction would consist of an all-inclusive assessment of gene expression in the liver. The purpose of this study was to perform a comprehensive analysis of gene expression in the livers of mice treated with ethanol by means of intragastric infusion. METHODS: An ethanol- or glucose enriched liquid diet was fed to animals for 4 weeks via a long-term gastrostomy catheter. The animals were killed and plasma alanine:2-oxoglutarate aminotransferase (ALT) assay, liver histology and total RNA analysis by microarray gene technology were performed. RESULTS: Alcohol increased ALT, induced steatosis, necrosis and inflammation. A total of 12,423 genes were analyzed for expression out of which 4867 were expressed by the liver. Alcohol repressed expression of 11 genes, induced expression of 13 genes, and up- or down regulated expression of 44 and 42 genes >2-fold, respectively. Gene expression analysis identified several genes that have not previously been tested for alcohol effects. CONCLUSIONS: This study: (i) expands the knowledge of mechanism(s) of action of ethanol; (ii) indicates novel pathways of ethanol action on the liver, and (iii) illustrates the utility of microarray gene analysis in hepatology research. PMID- 14739092 TI - Weight gain after transjugular intrahepatic portosystemic shunt is associated with improvement in body composition in malnourished patients with cirrhosis and hypermetabolism. AB - BACKGROUND/AIMS: To search for changes in body composition and energy metabolism associated with the repeatedly observed weight gain of cirrhotic patients after portosystemic shunting. METHODS: Twenty-one patients were studied prospectively before and 6 and 12 months after transjugular intrahepatic portosystemic shunt (TIPS) to assess body cell mass by two independent methods (total body potassium counting: body cell mass determined by TBP, BCMTBP, bioelectric impedance analysis: body cell mass determined by BIA, BCMBIA), muscle mass (anthropometry), resting energy expenditure (REECALO) by indirect calorimetry, and nutritional intake by dietary recall analysis. RESULTS: Prior to TIPS patients were hypermetabolic in terms of measured vs. predicted REE (REECALO median 1423 (range 1164-1838) vs. REEPRED 1279 (1067-1687) kcal; P<0.05) and their body cell mass was lower (19.1 (10.9-33.4) vs. 31.7 (16.8-47.1) kg; P=0.001). After TIPS body cell mass (BCMBIA) increased to 23.5 (12.7-44.3) (P<0.025) and 25.7 (14.2-39.7) kg (P=0.05) at 6 and 12 months after TIPS and this was confirmed by total potassium counting (BCMTBP before TIPS: 18.8 (10.6-26.7) vs. 22.4 (12.9-28.5) kg at 6 months; P<0.01). Hypermetabolism persisted throughout the study period. Energy and protein intake increased significantly by 26 and 33%. CONCLUSIONS: An increase of prognostically relevant variables body cell and muscle mass contributes to the weight gain after TIPS in malnourished patients with cirrhosis and hypermetabolism. PMID- 14739093 TI - Chronic inhibition of nitric oxide increases the collateral vascular responsiveness to vasopressin in portal hypertensive rats. AB - BACKGROUND/AIMS: Nitric oxide (NO), a potent vasodilator, plays a significant role in the vascular hyposensitivity to vasoconstrictors related to portal hypertension. Chronic NO inhibition ameliorates portal-systemic collaterals in portal hypertensive rats. This study investigated whether chronic NO inhibition by NG-nitro-L-arginine methyl ester (L-NAME) improves the portal-systemic collateral vascular responsiveness to arginine vasopressin (AVP) in portal hypertensive rats. METHODS: Partially portal vein-ligated (PVL) rats received L NAME in tap water (approximately 25 mg/kg per day) or tap water only (control) since 2 days prior to until 7 days after PVL. Mean arterial pressure was measured on the 8th day. By in situ perfusion model, different concentrations of AVP (10( 10)-10(-7) M) with a constant flow rate (20 ml/min) were applied to assess the perfusion pressure of collateral vessels. In another series, perfusion with different flow rates (5-30 ml/min) was used to obtain flow-pressure curves: the slopes represent collateral vascular resistances and higher resistances indicate less collaterals. RESULTS: Mean arterial pressure was higher in the L-NAME treated group than that of the control group (P<0.05). As compared with the controls, L-NAME-treated rats achieved significantly higher perfusion pressures in response to AVP. In addition, chronic L-NAME treatment also induced an increase of collateral vascular resistance, suggesting the attenuation of portal systemic shunting. CONCLUSIONS: Chronic NO inhibition ameliorates portal-systemic shunting and improves the collateral vascular responsiveness to AVP in portal hypertensive rats. PMID- 14739094 TI - Acute non-selective beta-adrenergic blockade reduces prolonged frequency-adjusted Q-T interval (QTc) in patients with cirrhosis. AB - BACKGROUND/AIMS: Earlier studies have shown a prolonged frequency-adjusted Q-T interval (QTc>0.440 s(1/2)) in a substantial fraction of patients with cirrhosis. The effect of beta-blockade on QTc is unknown, and its determination was the aim of the study. METHODS: Seventeen patients with cirrhosis received 80 mg propranolol orally during a haemodynamic investigation with measurements at baseline and 90 min after propranolol ingestion. RESULTS: Beta-blockade reduced cardiac output (-21%, P<0.001), heart rate (-20%, P<0.001), and the hepatic venous pressure gradient (HVPG, -17%, P<0.02). The mean QTc=0.460 s(1/2) was prolonged compared to 0.410 s(1/2) in age-matched controls (P<0.01). Whereas QTc decreased during beta-blockade in the cirrhotic patients (from 0.460 to 0.440 s(1/2), P<0.01), no effect was found in the subgroup with normal QTc (0.429 vs. 0.422 s(1/2), ns), and a reduction was seen in the patients with prolonged QTc (from 0.488 to 0.456 s(1/2), P<0.01). The percentage decrease in QTc was related to the reduction in HVPG (r=0.48, P=0.03) and cardiac output (r=0.56, P=0.02). CONCLUSIONS: Acute non-selective beta-blockade reduces prolonged QTc towards normal values in patients with cirrhosis. The clinical significance of QTc reduction in arrhythmia is a topic for future research. PMID- 14739095 TI - Systemic inflammatory response exacerbates the neuropsychological effects of induced hyperammonemia in cirrhosis. AB - BACKGROUND/AIMS: Studies in acute liver failure show correlation between evidence of a systemic inflammatory response syndrome (SIRS) and progression of hepatic encephalopathy (HE). We tested the hypothesis that SIRS mediators, such as nitric oxide and proinflammatory cytokines, may exacerbate the neuropsychological effects of hyperammonemia in cirrhosis. METHODS: Ten patients with cirrhosis were studied, 24-36 h after admission with clinical evidence of infection, and following its resolution. Hyperammonemia was induced by oral administration of an amino-acid (aa) solution mimicking hemoglobin composition. Inflammatory mediators, nitrate/nitrite, ammonia, aa profiles and a battery of neuropsychological tests were measured. RESULTS: The hyperammonemia generated in response to the aa solution was similar prior to, and after resolution, of the inflammation (P=0.77). With treatment of the infection there were significant reductions in white blood cell count (WBC), C-reactive protein (CRP), nitrate/nitrite, interleukin-6, interleukin-1beta and tumour necrosis factor alpha. Induced hyperammonemia resulted in significant worsening of the neuropsychological scores when patients showed evidence of SIRS but not after its resolution. CONCLUSIONS: The significant deterioration of neuropsychological test scores following induced hyperammonemia during the inflammatory state, but not after its resolution, suggests that the inflammation and its mediators may be important in modulating the cerebral effect of ammonia in liver disease. PMID- 14739096 TI - Commitment of bone marrow cells to hepatic stellate cells in mouse. AB - BACKGROUND/AIMS: Recently, several cells found within the liver have been reported to derive from bone marrow (BM). This study sought to examine the commitment of BM cells to hepatic stellate cell (HSC) lineage in mouse liver. METHODS: We transplanted BM cells from green fluorescent protein (GFP) transgenic mice into age-matched C57BL/J mice. Hepatic nonparenchymal cells were isolated from the livers of BM-transplanted mice using density gradient centrifugation with Nycodenz. The expression of lineage markers by the isolated cells was evaluated by RT-PCR and immunostaining. We then examined the histology of liver tissues obtained from BM-transplanted mice with and without carbon tetrachloride induced injury. RESULTS: GFP-expressing cells with intracytoplasmic lipid droplets comprised 33.4 +/- 2.3% of the cells isolated by density gradient centrifugation. These cells expressed the HSC lineage markers, such as desmin and glial fibrillary acidic protein (GFAP), by both RT-PCR and immunostaining. During a 7-day culture, GFP-positive cells began to express alpha-smooth muscle actin, a marker of activated HSC. In the liver of BM-transplanted mice, GFP-positive nonparenchymal cells expressed GFAP and extended their process around hepatocytes. Upon liver injury, these cells also co-expressed desmin and alpha smooth muscle actin. CONCLUSIONS: Nonparenchymal cells, derived from transplanted BM, acquired HSC characteristics in both quiescent and activated states. PMID- 14739097 TI - Searching for common stem cells of the hepatic and hematopoietic systems in the human fetal liver: CD34+ cytokeratin 7/8+ cells express markers for stellate cells. AB - BACKGROUND/AIMS: The hematopoietic and hepatic systems are intertwined in the liver during fetal life. Cells expressing the hematopoietic stem cell marker CD34 and cytokeratin 7/8 (CK7/8) are hypothesized to be common stem cells for the hematopoietic and hepatic systems. Our aim was to determine if human fetal liver cells expressing CD34 and CK7/8 represent a common stem cell for both the hematopoietic and hepatic systems. METHODS: CD34+CK7/8+ cells from midgestation livers were analyzed for the expression of various markers by flow cytometry and isolated based on their expression of CD34, nerve growth factor receptor (NGFR) and lack of CD45 expression. CD34+CD38- hematopoietic stem cells were also isolated and cultured in the presence of various hepatopoietins. RESULTS: CD34+CK7/8+ cells comprised 3.4-8.5% of the erythrocyte-depleted liver. CD34+CK7/8+ cells had unique light-scatter properties compared to hematopoietic precursors and did not express most markers associated with hematopoietic cells. They did stain with CD13, CD59, NGFR, desmin and alpha-smooth muscle actin. In culture, these cells had a stellate appearance. Cultured hematopoietic stem cells failed to generate hepatocytes. CONCLUSIONS: CD34+CK7/8+ cells are not common stem cells but rather appear to be hepatic stellate cells. A link between the hematopoietic and hepatic systems during fetal life requires further investigation. PMID- 14739098 TI - Halofuginone, an inhibitor of collagen synthesis by rat stellate cells, stimulates insulin-like growth factor binding protein-1 synthesis by hepatocytes. AB - BACKGROUND/AIMS: Halofuginone, an inhibitor of collagen synthesis, prevented and caused resolution of established hepatic fibrosis. A genomic approach in vivo was used to search for additional genes responsible for halofuginone mode of action. METHODS: Fibrosis was induced in rats by thioacetamide (TAA) and evaluated by collagen type I gene expression and the levels of collagen, tissue inhibitors of metalloproteinases-2 and smooth-muscle actin. Halofuginone was given in the diet. cDNA from liver biopsies was hybridized on Atlas arrays comprising of 588 genes. The results were confirmed by Northern blots and in situ hybridization. RESULTS: Insulin-like growth factor binding protein-1 (IGFBP-1) was one of the 13 genes differentially expressed in the fibrotic liver after halofuginone treatment. After 2 and 4 weeks, halofuginone prevented the TAA-induced down-regulation of IGFBP-1 gene expression. Halofuginone also prevented the TAA-dependent changes in IGFBP-3 gene expression. Halofuginone affected IGFBP-1 synthesis in rat hepatocytes and cells of hepatocyte origin and caused time- and dose-dependent increases in the IGFBP-1 gene expression and synthesis by HepG2 cells. The IGFBP 1 secreted by HepG2-inhibited stellate cell motility. CONCLUSIONS: Halofuginone is an anti-fibrotic drug that inhibits collagen synthesis by stellate cells and preventing alteration in the synthesis of IGFBPs by hepatic cells. PMID- 14739099 TI - Sodium butyrate enhances Fas-mediated apoptosis of human hepatoma cells. AB - BACKGROUND/AIMS: Human hepatoma cells have been reported to be resistant to Fas mediated apoptosis. Sodium butyrate (SB) induced apoptosis of several cancer cells. We investigated the effects of SB on Fas-mediated apoptosis of hepatoma cells. METHODS: In hepatoma cells (HuH-6, HuH-7, Hep-G2, and PLC/PRF/5), susceptibility to Fas-mediated apoptosis and Fas expression were assessed. Caspase-3 activation and cell cycle progression were evaluated in HuH-6. A cDNA microarray assay was performed to screen the changes in the expression of mRNAs. RESULTS: Pretreatment with SB caused an enhancement of the sensitivity to anti Fas-mediated cytotoxicity, though it did not increase the expression of Fas. The cDNA microarray assay revealed up-regulation of pro-apoptotic Bik, Bak, Bid and c Jun N-terminal protein kinase-1, and down-regulation of anti-apoptotic Bag-1 and cellular Fas-associated death domain-like interleukin-1beta-converting enzyme inhibitor protein. In some molecules, expression of the proteins was confirmed by Western blotting. An increase in truncated-Bid accompanying the reduction in Bid was also observed. CONCLUSIONS: SB enhances the susceptibility of hepatoma cells to anti-Fas-mediated cytotoxicity by altering the mRNA and protein expression and/or the activation status of proteins that could be involved in the Fas signaling pathway. SB may have an important role in the elimination of hepatoma cells. PMID- 14739100 TI - Involvement of natural killer T cells and granulocytes in the inflammation induced by partial hepatectomy. AB - BACKGROUND/AIMS: Natural killer T (NKT) cells are present in the liver of mice. We examined whether NKT cells and other leukocytes were associated with hepatic inflammation after partial hepatectomy. METHODS: Approximately 70% of the liver was removed from mice using the method described by Higgins and Anderson. RESULTS: Partial hepatectomy induced the expansion of NKT cells in the liver and the elevation of transaminase. These responses were completely suppressed by the administration of tacrolimus. NKT cell-deficient mice showed a decreased level of transaminase after partial hepatectomy. Perforin (-/-) mice showed an elevation of transaminase while B6-gld/gld mice (Fas ligand-) showed a decreased elevation of transaminase. In TAP-1(-/-) mice which lacked CD8+ cytotoxic T cells, inflammation remained at a normal level after partial hepatectomy. Since NKT cell deficient mice showed up to 50% decrease in the level of inflammation, we examined the association of other leukocytes with the remaining inflammation. The number and proportion of granulocytes were increased by partial hepatectomy. CONCLUSIONS: Both NKT cells and granulocytes participated in the hepatic inflammation after partial hepatectomy. The function of NKT cells, but not of granulocytes, was found to be sensitive to the immunosuppressive effect of tacrolimus. PMID- 14739101 TI - Herpes simplex virus thymidine kinase-mediated suicide gene therapy for hepatocellular carcinoma using HIV-1-derived lentiviral vectors. AB - BACKGROUND/AIMS: Gene therapy is a promising approach for treatment of hepatocellular carcinoma (HCC). However, transduction of non-tumoral hepatocytes may lead to severe hepatitis when using suicide gene therapy approaches. The aim of our study was to evaluate the gene transfer efficiency into HCC cells and normal hepatocytes using human immunodeficiency virus (HIV)-derived lentiviral vectors in vitro and in vivo. METHODS: Lentiviral vectors encoding for the LacZ gene or the fusion gene HSV-Tk/GFP were tested in vitro in human HCC cells and human hepatocytes in primary culture and in vivo in a chemically induced rat model of HCC. RESULTS: We show that HIV-1-derived lentiviral vectors are efficient in transducing HCC cells in vitro and in vivo. No significant transduction of non-tumorous hepatocytes was observed in vivo whatever the route of administration used. Measurement of tumor growth following direct intratumoral injection of a lentiviral vector containing the HSV-Tk gene and GCV treatment showed a strong antitumoral efficacy in the absence of normal liver toxicity. CONCLUSIONS: These observations suggest that lentiviral vectors allow an antitumoral effect with low liver toxicity when using suicide gene therapy approach and could be efficient tools for HCC gene therapy. PMID- 14739102 TI - Primary liver carcinoma of intermediate (hepatocyte-cholangiocyte) phenotype. AB - BACKGROUND/AIMS: Recent evidence of hepatic progenitor cells with the bipotential to differentiate into hepatocytes and cholangiocytes gives rise to the suggestion that primary hepatic carcinomas with features intermediate between hepatocellular carcinoma (HCC) and cholangiocarcinoma (CC) may originate from hepatic progenitor cells. METHODS: Fifty-four cases of primary liver carcinomas were selected and an immunohistochemical analysis was performed using hepatocytic markers (alpha fetoprotein, hepatocyte), cholangiocytic markers (carcinoembryonic antigen, cytokeratin 19) and progenitor cell marker (c-kit). RESULTS: Thirteen cases designated 'intermediate' carcinomas demonstrated strands/trabeculae of small, uniform, round-to-oval cells with scanty cytoplasm and hyperchromatic nuclei embedded within a thick desmoplastic stroma. Six were designated transitional type combined hepatocellular-cholangiocarcinoma (CHC). Ten were named HCC small cell type, demonstrating similar features to typical HCC, but composed of smaller cells. Simultaneous expression of hepatocytic and cholangiocytic markers was demonstrated in 8/13 (61.5%), 4/6 (66.7%), and 3/10 (30%) cases of intermediate carcinomas, transitional CHCs, and HCC small cell type, respectively, and c-kit expression was noted in 10/13 (76.9%), 4/6 (66.7%) and 7/10 (70%) cases, in the same order. CONCLUSIONS: Intermediate carcinoma may be a distinct type of primary liver carcinoma, morphologically and phenotypically intermediate between HCC and CC, which originates from transformed hepatic progenitor cells. PMID- 14739103 TI - Effect of vascular endothelial growth factor on functional recovery after hepatectomy in lean and obese mice. AB - BACKGROUND/AIMS: Liver regeneration is dependent upon coordinated proliferation of hepatocytes and endothelial cells. Vascular endothelial growth factor (VEGF) promotes angiogenesis. Hepatic steatosis delays regeneration and increases liver resection morbidity. We hypothesized that VEGF overexpression stimulates hepatic regeneration. METHODS: Recombinant adenovirus expressing human VEGF165 or adenovirus control-vector (LacZ) were administered before 2/3 hepatectomy in lean and ob/ob mice. Galactose elimination capacity, a quantitative liver function test, was repeatedly measured before and after hepatectomy. Expression of VEGF receptors (flt1, flk1), endoglin and hypoxia inducible factor-1alpha (HIF-1alpha) was assessed by quantitative RT-PCR and for endoglin also by immunohistochemistry. RESULTS: After 2/3 hepatectomy, VEGF gene transfer increased galactose elimination capacity in lean and ob/ob mice. HIF-1alpha, endoglin and VEGF receptor mRNA increased during regeneration in lean but not in obese mice. Staining of endothelial cells by endoglin immunohistochemistry returned to baseline reactivity in lean mice by day 6 and remained decreased in ob/ob mice. VEGF treatment decreased HIF-1alpha and increased flk1 response in lean mice. CONCLUSIONS: Hepatic resection elicits an angiogenic response in the remnant liver, which is impaired in case of steatosis. Adenovirus-mediated transfer of VEGF hastens functional hepatic recovery in lean, and more importantly also, in obese mice after partial hepatectomy. PMID- 14739104 TI - Analysis of T cell responses against hepatitis C virus genotype 4 in Egypt. AB - BACKGROUND/AIMS: Hepatitis C virus (HCV) remains the most common cause of chronic liver disease in Egypt. Despite the high prevalence of HCV and Schistosoma mansoni (S. mansoni) in Egypt, the effect of co-infection on the immune response against HCV genotype 4a has not been extensively examined. METHODS: We evaluated the HCV 4a-specific responses against the core and non-structural 5B proteins in chronic HCV with or without S. mansoni co-infection in 38 volunteers from Egypt. RESULTS: HCV 4a-specific responses were detected in 8/15 and 13/23 individuals with HCV alone or with concomitant schistosomiasis, respectively. Despite the alteration in the Th1 cytokine profile caused by schistosomiasis, the overall immune response rate against HCV was not affected (P=0.11). Seven individuals demonstrated HCV-specific responses against conserved regions of the Core that were previously identified for genotypes 1, 2 and 3 despite differences in HLA class I distribution. CONCLUSIONS: Egyptian patients infected with HCV genotype 4 can mount HCV-specific T cell responses, both CD4+ and CD8+ T cell-mediated, despite the prevalence of concomitant schistosomiasis. These findings suggest that S. mansoni co-infection may not represent a major obstacle to developing an HCV vaccine in this population. PMID- 14739105 TI - Comparing the public health burden of chronic hepatitis C and HIV infection in France. AB - BACKGROUND/AIMS: In France, the prevalence of hepatitis C virus (HCV) exceeds that of HIV, but in the absence of treatment, HIV infection progresses more rapidly than HCV. More HIV-infected patients, however, have received treatment. Using reported public health data in France and natural history models, we applied the back-calculation method to project future mortality from HCV and HIV incorporating current therapies. METHODS: The HCV model was based on literature data for the natural history of HCV and reports of hepatocellular carcinoma mortality. The HIV model used estimates from the French Hospital Database on HIV and reported AIDS cases and deaths. RESULTS: Peak annual mortality from HIV at 5000 occurred in 1994 and was 1000 in 1998, but HCV mortality likely increased through the 1990s and reached 3000 in 1998. Considering only HCV infections occurring until 1998 and currently available therapy, our model suggested that annual HCV-related mortality would continue to rise and would reach 4500 deaths in 2022. In contrast, AIDS-related deaths began to decrease in 1997. CONCLUSIONS: The public health burden of HCV is likely on the rise, while the burden of HIV, given the fairly widespread use of effective medications, may be on the decline. These results may help health policymakers in planning their responses to these epidemics. PMID- 14739106 TI - Infection, inflammation and hepatic encephalopathy, synergism redefined. PMID- 14739107 TI - On the origin of stellate cells: mesodermal, endodermal or neuro-ectodermal? PMID- 14739108 TI - Sensitizing to apoptosis--sharpening the medical sword. PMID- 14739109 TI - The promise of lentiviral gene therapy for liver cancer. PMID- 14739110 TI - Extrahepatic disease manifestations of HCV infection: some current issues. PMID- 14739111 TI - Erythropoietic protophyria in an adolescent female. PMID- 14739112 TI - Radiofrequency ablation for life-threatening ruptured hepatocellular carcinoma. PMID- 14739113 TI - Diagnosis of biliary duct cysts in transgenic mice expressing the hepatitis B virus X-protein. PMID- 14739114 TI - Ineffectiveness of hepatitis B vaccination in lamivudine-treated cirrhotics. PMID- 14739118 TI - Changes in dietary fat intake alter plasma levels of oxidized low-density lipoprotein and lipoprotein(a). AB - OBJECTIVE: To assess the effects of dietary modifications on oxidized low-density lipoprotein (LDL). METHODS AND RESULTS: Thirty-seven healthy women were fed two diets. Both diets contained a reduced amount of total and saturated fat. In addition, one diet was low in vegetables and the other was high in vegetables, berries, and fruit. The dietary intake of total fat was 70 g per day at baseline and decreased to 56 g (low-fat, low-vegetable diet) and to 59 g (low-fat, high vegetable diet). The saturated fat intake decreased from 28 g to 20 g and to 19 g, and the amount of polyunsaturated fat intake increased from 11 g to 13 g and to 19 g (baseline; low-fat, low-vegetable; low-fat, high-vegetable; respectively). The amount of oxidized LDL in plasma was determined as the content of oxidized phospholipid per ApoB-100 using a monoclonal antibody EO6 (OxLDL EO6). The median plasma OxLDL-EO6 increased by 27% (P<0.01) in response to the low-fat, low-vegetable diet and 19% (P<0.01) in response to the low-fat, high vegetable diet. Also, the Lp(a) concentration was increased by 7% (P<0.01) and 9% (P=0.01), respectively. CONCLUSIONS: Alterations in the dietary fat intake resulted in increased plasma concentrations of lipoprotein(a) and OxLDL-EO6. PMID- 14739119 TI - Mouse models of abdominal aortic aneurysms. AB - Many mouse models of abdominal aortic aneurysms have been developed that use a diverse array of methods for producing the disease, including genetic manipulation and chemical induction. These models could provide insight into potential mechanisms in the development of this disease. Although experimental studies on abdominal aortic aneurysms (AAAs) have used a variety of mammalian and avian approaches, there is an increasing reliance on the use of mice. The models recapitulate some facets of the human disease including medial degeneration, inflammation, thrombus formation, and rupture. Most of the mouse models of AAA are evoked either by genetically defined approaches or by chemical means. The genetic approaches are spontaneous and engineered mutations. These include defects in extracellular matrix maturation, increased degradation of elastin and collagen, aberrant cholesterol homeostasis, and enhanced production of angiotensin peptides. The chemical approaches include the intraluminal infusion of elastase, periaortic incubations of calcium chloride, and subcutaneous infusion of AngII. A common feature of these models is the reduction of AAA incidence and severity by the prophylactic administration of matrix metalloproteinase (MMP) inhibitors or genetically engineered deficiencies of specific members of this proteolytic protein family. The validation of mouse models of AAAs will provide insight into the mechanisms of progression of the human disease. PMID- 14739120 TI - Deleterious impact of "high normal" glucose levels and other metabolic syndrome components on arterial endothelial function and intima-media thickness in apparently healthy Chinese subjects: the CATHAY study. AB - OBJECTIVE: Endothelial vasodilator dysfunction and carotid intima-media thickening are useful surrogate markers of cardiovascular disease, a major cause of morbidity and mortality in type 2 diabetic patients. However, because most studies reporting the relationships between endothelial function, intima-media thickness (IMT), and hyperglycemia have compared diabetic patients with healthy controls, we report their relationship with glycemia as a continuum. METHODS AND RESULTS: Brachial artery endothelial function (flow-mediated dilatation [FMD]) and carotid IMT were measured noninvasively by high-resolution ultrasound B-mode imaging in 228 apparently healthy Chinese subjects recruited from Hong Kong and Macau. FMD and IMT were significantly associated with increasing levels of glycemia, particularly in the "high normal" glycemic range, with IMT increasing and endothelium-independent dilatation decreasing linearly across the glucose tertiles, and endothelium-dependent dilatation significantly lower in the upper glucose tertile compared with the other 2 groups (P<0.01). Using multiple linear regression, fasting glucose level was identified as an independent predictor of each of these markers of vascular function (P<0.004). Additionally, other conventional cardiovascular risk factors, including obesity, blood pressure, and an adverse lipid profile, were also related to levels of glycemia (P<0.05), further contributing to impaired vascular function. CONCLUSIONS: Increasing levels of glycemia and the coexistence of other cardiovascular risk factors in apparently healthy subjects are adversely associated with arterial endothelial dysfunction and intima-media thickening. PMID- 14739121 TI - Role of bone marrow-derived progenitor cells in cuff-induced vascular injury in mice. AB - OBJECTIVE: Arterial injury results in vascular remodeling associated with proliferation and migration of smooth muscle cells (SMCs) and the development of intimal hyperplasia, which is a critical component of restenosis after angioplasty of human coronary arteries and an important feature of atherosclerotic lesions. However, the origin of SMCs and other cells in the development of vascular remodeling is not yet fully understood. METHODS AND RESULTS: We utilized a cuff-induced vascular injury model after transplantation of the bone marrow (BM) from green fluorescent protein (GFP)-transgenic mice. We found that macrophages were major cells recruited to the adventitia of the vascular injury lesion along with SMCs and endothelial cells (ECs). While investigating whether those cells are derived from the donor, we found that most of the macrophages were GFP-positive, and some of the SMCs and ECs were also GFP positive. Administration of the anti-c-fms antibody resulted in a marked decrease in macrophages and a relative increase of SMCs, while administration of antibodies against the platelet-derived growth factor receptor-beta caused a prominent decrease in SMCs and a relative increase in macrophages. CONCLUSIONS: The current study indicates that BM-derived cells play an important role in vascular injury, and that differentiation of macrophages and SMCs might be dependent on each other. PMID- 14739122 TI - Monocyte chemoattractant protein-1 is an essential inflammatory mediator in angiotensin II-induced progression of established atherosclerosis in hypercholesterolemic mice. AB - OBJECTIVE: Chronic inflammatory processes might be involved in the progression and destabilization of atherosclerotic plaques. Therefore, identification of the mechanism underlying arterial inflammatory function might lead to the development of novel therapeutic strategies. Angiotensin II (AngII) is implicated in atherogenesis by activating the vascular inflammation system, mainly through monocyte chemotaxis. Therefore, we hypothesized that AngII increases plaque size and promotes destabilization of established atheromas by activating the monocyte chemoattractant protein-1 (MCP-1) pathway. METHODS AND RESULTS: We report here that 4-week infusion of AngII not only increased plaque size but also induced a destabilization phenotype (ie, increased macrophages and lipids and decreased collagen and smooth muscle cells) of pre-existing atherosclerotic lesions of hypercholesterolemic mice. AngII also enhanced the gene expression of inflammatory cytokines (TNFalpha, IL-6, etc.) and chemokines (MCP-1, CCR2, etc). Blockade of MCP-1, by transfecting the deletion mutant of the human MCP-1 gene into the skeletal muscles, limited AngII-induced progression and destabilization of established atherosclerotic lesions and suppressed the induction of proinflammatory genes. CONCLUSIONS: These data suggest that MCP-1 functions as a central inflammatory mediator in the AngII-induced progression and changes in plaque composition of established atheroma. PMID- 14739123 TI - Antioxidative activity of HDL particle subspecies is impaired in hyperalphalipoproteinemia: relevance of enzymatic and physicochemical properties. AB - OBJECTIVE: Hyperalphalipoproteinemia (HALP) is characterized by elevated plasma levels of high-density lipoprotein (HDL) particles with altered composition, metabolism, and function. The impact of such modification on antioxidative activities of HDL subfractions is indeterminate. METHODS AND RESULTS: Gradient fractionation revealed that buoyant HDL2b and 2a and small dense HDL3b and 3c levels were elevated up to 2.0-fold in HALP subjects (n=9; mean plasma HDL cholesterol, 79 mg/dL) with low hepatic lipase activity. HDL2a, 3a, 3b, and 3c displayed lower specific antioxidative activity (sAA) during low-density lipoprotein (LDL) oxidation (-15% to -86%, on a unit particle mass basis) than their normolipidemic counterparts (n=13). LDL oxidation was delayed by control HDL3a, 3b, and 3c (up to -79%) but specifically by HDL3c (-54%) in HALP. Paraoxonase activity was deficient in all HALP HDL subfractions. Paraoxonase, PAF AH, and LCAT activities together accounted for approximately 50% of variation in sAA. Abnormal chemical composition of HDL3b and 3c (cholesterol-deficient, triglyceride-enriched) in HALP was associated with impaired sAA. Systemic oxidative stress (as plasma 8-isoprostanes) tended to be elevated (1.5-fold) in HALP and negatively correlated with sAA (as TBARS). CONCLUSIONS: Intrinsic antioxidative activity of HDL subspecies is impaired in HALP, reflecting altered enzymatic and physicochemical properties. PMID- 14739124 TI - Inverse association between birth weight and C-reactive protein concentrations in the MIDSPAN Family Study. AB - OBJECTIVE: Inflammation markers and low birth weight each predict elevated risk of cardiovascular events and type 2 diabetes. However, potential associations between the low-grade inflammatory response as represented by C-reactive protein (CRP) concentrations and low birth weight have been sparsely examined. METHODS AND RESULTS: In the MIDSPAN Family Study, 1663 individuals had birth weight data and CRP concentrations measured as adults (age 30 to 59). The relationship between these parameters was examined after adjusting for factors known to influence CRP concentrations inclusive of age, body mass index, smoking, socio economic deprivation, and hormone use in women. After adjusting for potential confounders, there was a negative association between birth weight and CRP, whereby a 1-kg increase in birth weight is associated with a 10.7% decrease in CRP (95% CI: 3.0% to 17.8% decrease). There was no strong evidence that the effects differed in men and women (P=0.32). CONCLUSIONS: Low birth weight contributes to elevated CRP concentration in adult life. Future studies are required to determine to what extent this association reflects catch-up centile crossing, in utero programming, or genetic factors. PMID- 14739125 TI - Raising high-density lipoprotein in humans through inhibition of cholesteryl ester transfer protein: an initial multidose study of torcetrapib. AB - OBJECTIVE: The ability of the potent cholesteryl ester transfer protein (CETP) inhibitor torcetrapib (CP-529,414) to raise high-density lipoprotein cholesterol (HDL-C) levels in healthy young subjects was tested in this initial phase 1 multidose study. METHODS AND RESULTS: Five groups of 8 subjects each were randomized to placebo (n=2) or torcetrapib (n=6) at 10, 30, 60, and 120 mg daily and 120 mg twice daily for 14 days. Torcetrapib was well tolerated, with all treated subjects completing the study. The correlation of plasma drug levels with inhibition (EC50=43 nM) was as expected based on in vitro potency (IC50 approximately 50 nM), and increases in CETP mass were consistent with the proposed mechanism of inhibition. CETP inhibition increased with escalating dose, leading to elevations of HDL-C of 16% to 91%. Total plasma cholesterol did not change significantly because of a reduction in nonHDL-C, including a 21% to 42% lowering of low-density lipoprotein cholesterol at the higher doses. Apolipoprotein A-I and E were elevated 27% and 66%, respectively, and apoB was reduced 26% with 120 mg twice daily. Cholesteryl ester content decreased and triglyceride increased in the nonHDL plasma fraction, with contrasting changes occurring in HDL. CONCLUSIONS: These effects of CETP inhibition resemble those observed in partial CETP deficiency. This work serves as a prelude to further studies in subjects with low HDL, or combinations of dyslipidemia, in assessing the role of CETP in atherosclerosis. PMID- 14739126 TI - Vascular heme oxygenase-1 induction suppresses microvascular thrombus formation in vivo. AB - OBJECTIVE: By heme degradation, heme oxygenase-1 (HO-1) provides endogenous carbon monoxide and bilirubin, both of which play major roles in vascular biology. The current study aimed to examine whether induction of HO-1 and its byproducts modulate the process of microvascular thrombus formation in vivo. METHODS AND RESULTS: In individual microvessels of mouse cremaster muscle preparations, ferric chloride-induced thrombus formation was analyzed using intravital fluorescence microscopy. When mice were pretreated with an intraperitoneal injection of hemin, a HO-1 inducer, immunohistochemistry and Western blot protein analysis of cremaster muscle tissue displayed a marked induction of HO-1. In these animals, superfusion with ferric chloride solution induced arteriolar and venular thrombus formation, which, however, was significantly delayed when compared with thrombus formation in animals without HO 1 induction. The delay in thrombus formation in hemin-treated mice was completely blunted by tin protoporphyrin-IX, a HO-1 inhibitor, but not by copper protoporphyrin-IX, which does not inhibit the enzyme. Coadministration of the vitamin E analogue Trolox in HO-1-blocked animals almost completely restored the delay in thrombus formation, implying that, besides CO, the antioxidant HO pathway metabolite bilirubin mainly contributes to the antithrombotic property of HO-1. This was further supported by the fact that bilirubin was found as effective as hemin in delay of ferric chloride-induced thrombus formation. Animals with HO-1 induction revealed reduced P-selectin protein expression in cremaster muscle tissue, which most probably presented the molecular basis for delayed thrombus growth. CONCLUSIONS: Local induction of HO-1 activity may be of preventive and therapeutic value for clinical disorders with increased risk of thrombotic events. PMID- 14739127 TI - Inhibition of plasmin activity by tranexamic acid does not influence inflammatory pathways during human endotoxemia. AB - OBJECTIVE: Plasmin activates several proinflammatory pathways at the cellular level in vitro. Lipopolysaccharide (LPS) administration to healthy humans results in a rapid generation of plasmin activity, accompanied by activation of a number of inflammatory systems. METHODS AND RESULTS: To determine the role of early plasmin activity in LPS-induced inflammation in vivo, 16 healthy males received an intravenous bolus injection with LPS (from Escherichia coli, 4 ng/kg) directly preceded by a 30-minute intravenous infusion of tranexamic acid (2 g, n=8), a plasmin activation inhibitor, or placebo (n=8). LPS injection induced marked increases in the plasma levels of D-dimer and plasmin-alpha2-antiplasmin complexes, indicative of plasmin activation and generation, respectively, which were strongly attenuated by tranexamic acid (both P<0.01 versus placebo). However, tranexamic acid did not influence LPS-induced coagulation activation, granulocytosis, neutrophil activation (expression of CD11b, CD66b, and L selectin) or degranulation (plasma concentrations of elastase-alpha1-antitrypsin and bactericidal permeability-increasing protein), endothelial cell activation (plasma levels of von Willebrand factor and soluble E-selectin), or cytokine release. CONCLUSIONS: These data argue against a role of early plasmin generation in the subsequent activation of other inflammatory pathways during human endotoxemia. PMID- 14739128 TI - Blood cultures for community-acquired pneumonia: piecing together a mosaic for doing less. PMID- 14739129 TI - Vascular dysfunction in sleep apnea: a reversible link to cardiovascular disease? PMID- 14739130 TI - Obstructive sleep apnea and heart failure: two unhappy bedfellows. PMID- 14739131 TI - Natural history of adult-onset asthma: insights from model of occupational asthma. PMID- 14739132 TI - Modulation of alveolar fluid clearance by acute inflammation: the plot thickens. PMID- 14739133 TI - Fluoroquinolones in the treatment of tuberculosis: a study in mice. PMID- 14739134 TI - Ventilator-induced diaphragmatic dysfunction. PMID- 14739135 TI - Hyperglycemia, bronchial artery sclerosis, and lung function. PMID- 14739136 TI - Evidence is lacking for link between rhinitis and hypertension. PMID- 14739137 TI - Interferon-gamma toxicity in idiopathic pulmonary fibrosis. PMID- 14739138 TI - Progressive apoptotic cell death triggered by transient oxidative insult in H9c2 rat ventricular cells: a novel pattern of apoptosis and the mechanisms. AB - Many pathophysiological processes are associated with oxidative stress and progressive cell death. Oxidative stress is an apoptotic inducer that is known to cause rapid cell death. Here we show that a brief oxidative insult (5-min exposure to 400 microM H(2)O(2)), although it did not kill H9c2 rat ventricular cells during the exposure, triggered an intracellular death cascade leading to delayed time-dependent cell death starting from 1 h after the insult had been withdrawn, and this post-H(2)O(2) cell death cumulated gradually, reaching a maximum level 8 h after H(2)O(2) withdrawal. By comparison, sustained exposure to H(2)O(2) caused complete cell death within a narrow time frame (2 h). The time dependent post-H(2)O(2) cell death was typical of apoptosis, both morphologically (cell shrinkage and nuclear condensation) and biochemically (DNA fragmentation, extracellular exposure of phosphatidylserines, and caspase-3 activation). A dichlorofluorescein fluorescent signal showed a time-dependent endogenous increase of reactive oxygen species (ROS) production, which was almost abolished by inhibition of the mitochondrial electron transport chain. Application of antioxidants (vitamin E or DTT) before H(2)O(2) addition or after H(2)O(2) withdrawal prevented the H(2)O(2)-triggered progressive ROS production and apoptosis. Sequential appearance of events associated with activation of the mitochondrial death pathway was found, including progressive dissipation of mitochondrial membrane potential, cytochrome c release, and late activation of caspase-3. In conclusion, transient oxidative stress triggers an intrinsic program leading to self-sustained apoptosis in H9c2 cells via cumulative production of mitochondrial ROS and subsequent activation of the mitochondrial death pathway. This pattern of apoptosis may contribute to the progressive and long-lasting cell loss in some degenerative diseases. PMID- 14739139 TI - Autonomic regulation of calcium and potassium channels is oppositely modulated by microtubules in cardiac myocytes. AB - We recently showed that colchicine treatment of rat ventricular myocytes increases the L-type Ca2+ current (I(Ca)) and intracellular Ca2+ concentration ([Ca2+](i)) transients and interferes with adrenergic signaling. These actions were ascribed to adenylyl cyclase (AC) stimulation after G(s) activation by alpha,beta-tubulin. Colchicine depolymerizes microtubules into alpha,beta-tubulin dimers. This study analyzed muscarinic signals in myocytes with intact or depolymerized microtubules. Myocytes were loaded with the Ca2+ indicator fluo 3 and were field stimulated at 1 Hz or voltage clamped. In untreated cells, carbachol (CCh; 1 microM) induced ACh-activated K(+) current [I(K(ACh))], which happens via betagamma-subunits from the activation of G(i). Carbachol also reduced [Ca2+](i) transients and contractions. Once G(i) is activated by muscarinic agonist, the alpha(i)-subunit is released from the betagamma-subunits, but it is silent, and its inhibition of the AC/cAMP cascade, manifested by I(Ca) reduction, is not seen unless AC has been previously activated. In colchicine treated cells, CCh caused greater reductions of [Ca2+](i) transients and contractions than in untreated cells. The alpha(i)-subunit became effective in signaling through the AC/cAMP cascade and reduced I(Ca) without changing its voltage-dependence. Isoproterenol (Iso) regained its efficacy and reversed I(Ca) inhibition by CCh. Stimulation of I(Ca) by forskolin persisted in colchicine treated cells when Iso was ineffective. The effect of CCh on I(K(ACh)) was occluded in colchicine-treated cells. Colchicine treatment, per se, may increase I(K(ACh)) by betagamma-subunits released from G(s) to mask this effect of CCh. Microtubules suppress I(Ca) regulation by alpha(i); their disruption releases restraints that unmask muscarinic inhibition of I(Ca). Summarily, colchicine treatment reverses regulation of ventricular excitation-contraction coupling by autonomic agents. PMID- 14739140 TI - Inhibitory effects of galanin on evoked [Ca2+]i responses in cultured myenteric neurons. AB - Galanin modulates gastrointestinal motility by inhibiting the release of ACh from enteric neurons. It is, however, not known whether galanin also inhibits neuronal cholinergic transmission postsynaptically and whether galanin also reduces the action of other excitatory neurotransmitters. The aim of the present study was thus to investigate the effect of galanin on the evoked intracellular Ca(2+) concentration ([Ca(2+)](i)) responses in myenteric neurons. Cultured myenteric neurons from small intestine of adult guinea pigs were loaded with the Ca(2+) indicator fluo-3 AM, and the [Ca(2+)](i) responses following the application of different stimuli were quantified by confocal microscopy and expressed as a percentage of the response to high-K(+) solution (75 mM). Trains of electrical pulses (2 s, 10 Hz) were applied to stimulate the neuronal fibers before and after a 30-s superfusion with galanin (10(-6) M). Substance P (SP), 5-HT, 1,1 dimethyl-4-phenyl-piperazinium iodide (DMPP), and carbachol were used as direct postsynaptic stimuli (10(-5) M, 30 s) and were applied alone or after galanin perfusion. Galanin significantly reduced the responses induced by electrical fiber stimulation (43 +/- 2 to 35 +/- 3%, P = 0.01), SP (15.4 +/- 1 to 8.0 +/- 0.3%, P < 0.01), and 5-HT (26 +/- 2 to 21.4 +/- 1.5%, P < 0.05). On the contrary, galanin did not affect the responses induced by local application of DMPP and carbachol. We conclude that in cultured myenteric neurons, galanin inhibits the excitatory responses induced by electrical stimulation, SP, and 5-HT. Finally, the inhibitory effect of galanin on electrical stimulation, but not on DMPP- and carbachol-induced responses, suggests that, at least for the cholinergic component, galanin acts at the presynaptic level. PMID- 14739141 TI - Coactivation of capacitative calcium entry and L-type calcium channels in guinea pig gallbladder. AB - We have evaluated the presence of capacitative Ca(2+) entry (CCE) in guinea pig gallbladder smooth muscle (GBSM), including a possible relation with activation of L-type Ca(2+) channels. Changes in cytosolic Ca(2+) concentration induced by Ca(2+) entry were assessed by digital microfluorometry in isolated, fura 2-loaded GBSM cells. Application of thapsigargin, a specific inhibitor of the Ca(2+) store pump, induced a transient Ca(2+) release followed by sustained entry of extracellular Ca(2+). Depletion of the stores with thapsigargin, cyclopiazonic acid, ryanodine and caffeine, high levels of the Ca(2+)-mobilizing hormone cholecystokinin octapeptide, or simple removal of external Ca(2+) resulted in a sustained increase in Ca(2+) entry on subsequent reapplication of Ca(2+). This entry was attenuated by 2-aminoethoxydiphenylborane, L-type Ca(2+) channel blockade, pinacidil, and Gd(3+). Accumulation of the voltage-sensitive dye 3,3' dipentylcarbocyanine and direct intracellular recordings showed that depletion of the stores is sufficient for depolarization of the plasma membrane. Contractility studies in intact gallbladder muscle strips showed that CCE induced contractions. The CCE-evoked contraction was sensitive to 2-aminoethoxydiphenylborane, L-type Ca(2+) channel blockers, and Gd(3+). We conclude that, in GBSM, release of Ca(2+) from internal stores activates a CCE pathway and depolarizes plasma membrane, allowing coactivation of voltage-operated L-type Ca(2+) channels. This process may play a role in excitation-contraction coupling in GBSM. PMID- 14739142 TI - Salmonella typhimurium SipA-induced neutrophil transepithelial migration: involvement of a PKC-alpha-dependent signal transduction pathway. AB - Salmonella typhimurium elicits an intense proinflammatory response characterized by movement of polymorphonuclear neutrophils (PMN) across the epithelial barrier to the intestinal lumen. We previously showed that S. typhimurium, via the type III secretion system effector protein SipA, initiates an ADP-ribosylation factor 6- and phospholipase D-dependent lipid-signaling cascade that directs activation of protein kinase C (PKC) and subsequent transepithelial movement of PMN. Here we sought to determine the specific PKC isoforms that are induced by the S. typhimurium effector SipA in model intestinal epithelia and to link the functional consequences of these isoforms in the promotion of PMN transepithelial migration. In vitro kinase PKC activation assays performed on polarized monolayers of T84 cells revealed that S. typhimurium and recombinant SipA induced activation of PKC-alpha, -delta, and -epsilon. To elucidate which of these isoforms play a key role in mediating epithelial cell responses that lead to the observed PMN transepithelial migration, we used a variety of PKC inhibitors with different isoform selectivity profiles. Inhibitors selective for PKC-alpha (Go 6976 and 2,2',3,3',4,4'-hexahydroxyl-1,1'-biphenyl-6,6'-dimethanoldimethyl ether) markedly reduced S. typhimurium- and recombinant SipA-induced PMN transepithelial migration, whereas inhibitors to PKC-delta (rottlerin) or PKC-epsilon (V1-2) failed to exhibit a significant decrease in transepithelial movement of PMN. These results were confirmed biochemically and by immunofluorescence coupled to confocal microscopy. Our results are the first to show that the S. typhimurium effector protein SipA can activate multiple PKC isoforms, but only PKC-alpha is involved in the signal transduction cascade leading to PMN transepithelial migration. PMID- 14739143 TI - Calcium-switch technique and junctional permeability in native rabbit esophageal epithelium. AB - The Ca(2+)-switch technique was used to investigate the nature of the barrier governing (paracellular) permeability across the junctions of "native" rabbit esophageal epithelium. This was done by mounting esophageal epithelium in Ussing chambers to monitor transepithelial electrical resistance (R(T)), a marker of junctional permeability. When exposed to Ca(2+)-free Ringer solutions containing EDTA, R(T) declined approximately 35% below baseline over 2 h, and this decline reversed within 2 h by restoration of (1.2 mM) Ca(2+)-containing, normal Ringer solution ("Ca(2+)-switch technique"). Junctional resealing, i.e., increased R(T) on Ca(2+) replacement, was assessed by the Ca(2+)-switch technique and shown to be 1) specific for Ca(2+), with only Mn(2+) among substituted divalent cations yielding partial resealing; 2) a function of extracellular Ca(2+) levels because maneuvers (BAPTA/AM or A23187 exposure) to alter intracellular Ca(2+) had no effect; 3) dose dependent, requiring as a minimum > or =0.5 mM Ca(2+) and 1.2 mM Ca(2+) for optimization; and 4) independent of protein synthesis because it was not inhibited by cycloheximide. Resealing was also inhibited by luminal antibodies or synthetic peptides to the extracellular domain of E-cadherin. Immunohistochemistry revealed E-cadherin within all layers of stratum corneum in Ca(2+)-free but not Ca(2+)-containing solution. The present investigation documents, using the Ca(2+)-switch technique, that esophageal epithelial junctions contain a major Ca(2+)-dependent component and that this component reflects adhesion between the extracellular domains of E-cadherin containing a histidine-alanine-valine recognition sequence. PMID- 14739144 TI - Rapid induction of GATA transcription factors in developing mouse intestine following glucocorticoid administration. AB - In the developing intestine, transcription of alpha-glucosidase genes such as sucrase-isomaltase and trehalase is stimulated by glucocorticoid administration. The consequent increase of their respective mRNAs is characterized by a 12-h lag, suggesting that the response to glucocorticoids represents a secondary effect. We hypothesized that the primary response of the tissue to glucocorticoids includes induction of one or more intestinal transcription factors. To investigate this hypothesis, we identified a region in the mouse trehalase promoter (located at nucleotides -406 to -377 from the transcription start site) with potential binding sites for three transcription factors: Cdx-2, GATA, and C/EBP. Gel shifts were performed using labeled oligonucleotides from this region with nuclear extracts from jejunums of either control 8-day-old mouse pups or littermates treated with dexamethasone (DEX) 4 h before death. A specific shifted band was observed with DEX extracts but not with control extracts. Supershift assays indicated the presence of GATA-4 and GATA-6 but not GATA-5 nor Cdx-2, C/EBP alpha, C/EBP beta, or C/EBP delta. GATA binding was further implicated by competition studies with mutated oligonucleotides. Finally, Western blot analysis showed GATA-4 and GATA-6 proteins in DEX but not control nuclear extracts. For GATA-4, the same pattern was demonstrated with whole cell extracts and with the cytosol fraction. We conclude that expression of GATA-4 and GATA-6 proteins in the suckling mouse jejunum is stimulated by DEX. This novel finding constitutes an important first step in understanding the molecular mechanism of glucocorticoid action on the developing intestine. PMID- 14739145 TI - Inflammation of the cystic fibrosis mouse small intestine. AB - The CFTR null mouse [cystic fibrosis (CF) mouse] has a severe intestinal phenotype that serves as a model for CF-related growth deficiency, meconium ileus, and distal intestinal obstructive syndrome. DNA microarray analysis was used to investigate gene expression in the CF mouse small intestine. Sixty-one genes exhibited a statistically significant twofold or greater increase in expression, and 98 genes were downregulated twofold or greater. Of the upregulated genes, most were associated with inflammation and included markers for cells of the innate immune system (mast cells and neutrophils) and for acute phase genes (serum amyloid A and complement factors). The downregulated genes include 10 cytochrome P-450 genes; several are involved in lipid metabolism, and several are involved in various transport processes. Confirmation by quantitative RT-PCR showed gene expression was significantly increased for mast cell protease 2 (27-fold), hematopoietic cell transcript 1 (17-fold), serum amyloid A3 (2.9 fold), suppressor of cytokine signaling 3 (2.0-fold), leucine-rich alpha(2) glycoprotein (21-fold), resistin-like molecule-beta (49-fold), and Muclin (2.5 fold) and was significantly decreased for cytochrome P-450 4a10 (28-fold) and cubilin (114-fold). Immune cell infiltration was confirmed histologically by staining for mast cells and neutrophils. These data demonstrate that the CF intestine exhibits an inflammatory state with upregulation of components of the innate immune system. PMID- 14739146 TI - Human cytomegalovirus double resistance in a donor-positive/recipient-negative lung transplant patient with an impaired CD4-mediated specific immune response. AB - BACKGROUND: Emergence of human cytomegalovirus (HCMV) resistance to ganciclovir in solid-organ transplant recipients has been found to be mostly associated with primary HCMV infection. MATERIALS AND METHODS: The case of a donor positive/recipient-negative (D(+)/R(-)) lung transplant patient developing ganciclovir and cidofovir resistance is described. HCMV infection was monitored by weekly determination of antigenaemia, viraemia and DNAaemia. HCMV-specific CD4 cell immunity was determined by cytokine flow cytometry. The emergence of drug resistant HCMV strains was documented by sequencing of UL97 and UL54 genes of HCMV directly in blood samples. RESULTS: Following primary HCMV infection, the patient showed repeated reactivations for over a year, eventually resulting in the selection of a ganciclovir-resistant HCMV strain with a mutation in the UL97 gene product (A594V). Determination of HCMV-specific CD4 cell immunity showed a persistently impaired immune response. Subsequent foscarnet treatment allowed only transitory virus clearance from blood owing to renal toxicity. Further ganciclovir treatment induced a new mutation in both UL97 (H520Q) and UL54 (P522S) with final emergence of double resistance to both ganciclovir and cidofovir. The patient eventually died of lung failure. DISCUSSION: Determination of HCMV-specific CD4 cell immunity could be of help in predicting the emergence of drug-resistant strains in D(+)/R(-) transplant recipients. PMID- 14739147 TI - Overexpression of the multidrug efflux pump SmeDEF impairs Stenotrophomonas maltophilia physiology. AB - OBJECTIVES: The use of antibiotics for the treatment of infectious diseases has led to important changes in the structure of pathogenic bacterial populations. However, these changes could be buffered if the expression of antibiotic resistance genes were to lead to the counter-selection of antibiotic-resistant strains in antibiotic-free environments. To test the effect of antibiotic resistance on bacterial fitness, we analysed the effect of the overproduction of the multidrug efflux pump SmeDEF on the physiology of Stenotrophomonas maltophilia. SmeDEF confers resistance to antibiotics belonging to different structural families, and its overexpression is associated with an antibiotic resistance phenotype in clinical isolates of S. maltophilia. RESULTS: Two S. maltophilia isogenic strains were analysed: the wild-type strain D457 and strain D457R, which is a SmeDEF overproducer. In co-culture experiments, under non selective pressure the wild-type strain displaced the mutant strain D457R. Metabolic profiling showed that SmeDEF overproduction leads to several changes in S. maltophilia metabolism. Using a Dictyostelium discoideum model of bacterial virulence, we found overexpression of SmeDEF to be associated with a reduction in S. maltophilia virulence. CONCLUSIONS: Together, these data indicate that overexpression of the multidrug efflux pump SmeDEF causes a metabolic burden for S. maltophilia. PMID- 14739148 TI - Amphotericin B lipid complex versus no treatment in the secondary prophylaxis of visceral leishmaniasis in HIV-infected patients. AB - OBJECTIVES: Visceral leishmaniasis (VL) in HIV-positive patients is characterized by a chronic course with frequent relapse. The aim of this study was to evaluate the efficacy and safety of amphotericin B lipid complex (ABLC) in preventing VL relapses in HIV-infected patients. METHODS: This was a multicentre, open-label (with blinded centralized randomization), parallel, no-treatment, controlled clinical trial. HIV-infected patients, with at least one previous treated episode of VL and with negative bone marrow aspirate for Leishmania parasites prior to the study, were randomized to receive either ABLC 3 mg/kg/day every 21 days (ABLC) or no treatment (NT). Patients were followed-up every 9 weeks for up to 12 months, and the efficacy was measured as the proportion of patients remaining free (non-relapse) of VL at 1 year of follow-up. The primary analysis was performed on an intention-to-treat basis. RESULTS: One hundred and fifteen patients were screened, but only 17 were randomized: eight in the ABLC group and nine in the NT group. The intention-to-treat analysis of data showed 50% of patients remaining free of VL at 12 months of follow-up (95% CI = 15.7%, 84.3%) in the ABLC group, and 22.2% (95% CI = 2.8%, 60.0%) in the NT group. The non relapse odds ratio was 3.5 (95% CI = 0.30%, 52.0%) favouring ABLC. ABLC was well tolerated: patients only presented infusion-related mild adverse events. No patients from either group discontinued treatment or died during follow-up. CONCLUSIONS: ABLC, administered every 21 days for 12 months, is useful as secondary prophylaxis in preventing VL relapse in HIV-infected patients, and is well tolerated. PMID- 14739149 TI - Chemical and biological characterization of a polysaccharide biological response modifier from Aloe vera L. var. chinensis (Haw.) Berg. AB - Three purified polysaccharide fractions designated as PAC-I, PAC-II, and PAC-III were prepared from Aloe vera L. var. chinensis (Haw.) Berg. by membrane fractionation and gel filtration HPLC. The polysaccharide fractions had molecular weights of 10,000 kDa, 1300 kDa, and 470 kDa, respectively. The major sugar residue in the polysaccharide fractions is mannose, which was found to be 91.5% in PAC-I, 87.9% in PAC-II, and 53.7% in PAC-III. The protein contents in the polysaccharide fractions was undetectable. NMR study of PAC-I and PAC-II demonstrated the polysaccharides shared the same structure. The main skeletons of PAC-I and PAC-II are beta-(1-->4)-D linked mannose with acetylation at C-6 of manopyranosyl. The polysaccharide fractions stimulated peritoneal macrophages, splenic T and B cell proliferation, and activated these cells to secrete TNF alpha, IL-1 beta, INF-gamma, IL-2, and IL-6. The polysaccharides were nontoxic and exhibited potent indirect antitumor response in murine model. PAC-I, which had the highest mannose content and molecular weight, was found to be the most potent biological response modifier of the three fractions. Our results suggested that the potency of aloe polysaccharide fraction increases as mannose content and molecular weight of the polysaccharide fraction increase. PMID- 14739150 TI - Gene-altered islets for transplant: giant leap or small step? PMID- 14739151 TI - Brain aromatase: dyed-in-the-wool homosexuality. PMID- 14739153 TI - Endocrine-related resources from the National Institutes of Health. PMID- 14739152 TI - Adiponectin and HIV-lipodystrophy: taking HAART. PMID- 14739154 TI - National Hormone & Peptide Program (NHPP): new recombinant hormones, hypothalamic peptides, natural hormones, new antisera, expanded hormone Assay services available. PMID- 14739155 TI - Genome-wide scan identifies novel QTLs for cholesterol and LDL levels in F2[Dahl RxS]-intercross rats. AB - Hypercholesterolemia is a significant risk factor for coronary artery disease development. Genes influencing nonmonogenic hypercholesterolemia susceptibility in humans remain to be identified. Animal models are key investigative systems because major confounding variables such as diet, activity, and genetic background can be controlled. We performed a 121-marker, total genome-analysis of an F2[Dahl RxS]-intercross selected for contrasting parental strain susceptibilities for hyperlipidemia on regular rat diets at 6 months of age. Quantitative traits studied were plasma total cholesterol, triglyceride, HDL, and LDL levels adjusted for obesity. Genome-wide analysis of 200 F2-intercross male rats detects two QTLs with highly significant linkage for total cholesterol (TC) on chromosome (chr) 5-133.3 Mbp (LOD 5.8), and chr5-54.2 Mbp (LOD 4.8), and two QTLs with significant linkage for TC: on chromosome 8, chr8-60.4 Mbp (LOD 3.8), and chromosome 2, chr2-243.5 Mbp (LOD 3.4). A QTL for LDL with significant linkage is detected on chromosome 5, chr5-104 Mbp (LOD 3.7). These QTLs contribute from 7% to 12% of total trait variance, respectively, with Dahl-S allele effects resulting in increased TC and LDL levels consistent with hyperlipidemia susceptibility in the parental Dahl-S rat strain. Predicted QTL peaks do not coincide with previous genome scans. Human homologues of two TC-QTLs span genes listed in a LocusLink profile for cholesterol. Only suggestive loci were detected for HDL and total triglyceride levels. Altogether, the data demonstrates the contribution of multiple QTLs to hypercholesterolemia making a multipathway pathogenic framework imperative. QTL-peak candidate genes delineated are syntenic between rat and human genomes, increasing clinical relevance and mandating further study. PMID- 14739157 TI - Increased intimal hyperplasia and smooth muscle cell proliferation in transgenic mice with heparan sulfate-deficient perlecan. AB - Smooth muscle cell (SMC) proliferation is a critical process in vascular disease. Heparan sulfate (HS) proteoglycans inhibit SMC growth, but the role of endogenous counterparts in the vessel wall in control of SMC function is not known in detail. Perlecan is the major HS proteoglycans in SMC basement membranes and in vessel wall extracellular matrix (ECM). In this study, transgenic mice with HS deficient perlecan were analyzed with respect to vascular phenotype and intimal lesion formation. Furthermore, SMC cultures were established and characterized with respect to morphology, immunocytochemical features, proteoglycan synthesis, proliferative capacity, and ECM binding of basic fibroblast growth factor (FGF 2). In vitro, mutant SMCs formed basement membranes with perlecan core protein, but with decreased levels of HS, they showed diminished secretion of HS containing perlecan into the medium and a defective ECM-binding capacity of FGF 2. In vitro, mutant SMCs showed increased proliferation compared with wild-type cells, and in vivo, enhanced SMC proliferation and intimal hyperplasia were observed after flow cessation of the carotid artery in mutant mice. The results indicate that the endogenous HS side-chains of perlecan contribute to SMC growth control both in vitro and during intimal hyperplasia, possibly by sequestering heparin-binding mitogens such as FGF-2. PMID- 14739156 TI - Enhanced S-nitroso-albumin formation from inhaled NO during ischemia/reperfusion. AB - In the present study, we investigated whether inhaled nitric oxide (NO) was transported by plasma proteins, such as S-nitroso-albumin (SNO-Alb), in the feline circulation and whether this molecule delivers NO to the periphery under conditions of stress, specifically ischemia/reperfusion (I/R). A flow probe was interposed between the femoral and superior mesenteric artery for blood flow measurements, and a branch of the superior mesenteric vein was cannulated for arterial-venous sampling. In animals breathing room air, SNO-Alb was below detection level in arterial or venous blood. NO inhalation resulted in a significant arterial-venous gradient for SNO-Alb. Concomitant with this loss of SNO-Alb across the intestinal vasculature was an increase in nitrite (NO2-). However, this release of NO was not sufficient to alter intestinal blood flow. I/R during NO inhalation caused a very large increase in arterial SNO-Alb that permitted a 5-fold increase in SNO-Alb consumption and significant generation of NO2- within the postischemic intestinal vasculature. The increased SNO-Alb consumption was sufficient to dramatically improve intestinal blood flow. The very large burst of arterial SNO-Alb during I/R was completely blocked by the administration of superoxide dismutase, suggesting that oxidative stress contributed to the increased SNO-Alb formation. Our data suggest that inhaled NO can increase nitrosothiol production and these molecules may be a functional NO delivery system during cardiovascular disease. PMID- 14739158 TI - Muscarinic M2 receptor stimulation of Cav1.2b requires phosphatidylinositol 3 kinase, protein kinase C, and c-Src. AB - This study investigated regulation of L-type calcium channels (Cav1.2b) by acetylcholine (ACh) in rabbit portal vein myocytes. Whole-cell currents were recorded using 5 mmol/L barium as charge carrier. ACh (10 micromol/L) increased peak currents by 40%. This effect was not reversed by the selective muscarinic M3 receptor antagonist 4-DAMP (100 nmol/L) but was blocked by the M2 receptor antagonist methoctramine (5 micromol/L). The classical and novel protein kinase C (PKC) antagonist calphostin C (50 nmol/L) abolished ACh responses, whereas the classical PKC antagonist Go6976 (200 nmol/L) had no effect. ACh responses were also abolished by the phosphatidylinositol 3-kinase (PI3K) inhibitor LY294002 (20 micromol/L), by the c-Src inhibitor PP2 (10 micromol/L) (but not the inactive analogue PP3), and by dialyzing cells with an antibody to the G-protein subunit Gbetagamma. Cells dialyzed with c-Src had significantly greater currents than control cells. Current enhancement persisted in the presence of LY294002, suggesting that c-Src is downstream of PI3K. Phorbol 12,13-dibutyrate (PDBu, 0.1 micromol/L) increased currents by 74%. This effect was abolished by calphostin C and reduced by Go6976. The PDBu response was also reduced by PP2, and the PP2 insensitive component was blocked by Go6976. In summary, these data suggest that ACh enhances Cav1.2b currents via M2 receptors that couple sequentially to Gbetagamma, PI3K, a novel PKC, and c-Src. PDBu stimulates the novel PKC/c-Src pathway along with a second pathway that is independent of c-Src and involves a classical PKC. PMID- 14739159 TI - Calcineurin promotes the expression of monocyte chemoattractant protein-1 in vascular myocytes and mediates vascular inflammation. AB - Although the role of the calcineurin-dependent pathway in the development of cardiac hypertrophy has been intensively studied, little is known of its role in vascular inflammatory diseases such as atherosclerosis and restenosis after angioplasty. To help elucidate the role of calcineurin in vascular inflammation, we infected cultured vascular smooth muscle cells (VSMCs) with an adenovirus construct expressing a constitutively active mutant of calcineurin, and examined its effect on the expression of monocyte chemoattractant protein-1 (MCP-1). We also examined the role of calcineurin in vivo using a transluminal wire injury model of the rat femoral artery. Forced activation of calcineurin significantly increased the expression of MCP-1 both at the transcriptional and protein levels. Angiotensin II (Ang II) also significantly stimulated MCP-1 expression, and this increase was significantly inhibited by cyclosporin A (CyA). Constitutive activation of calcineurin stabilized MCP-1 mRNA without enhancing MCP-1 promoter activity. In accordance with the results, Ang II-induced increase of MCP-1 promoter activity was not suppressed by CyA. Ang II stabilized MCP-1 mRNA, and this effect of Ang II was diminished by CyA. CyA suppressed MCP-1 expression in the femoral artery after the transluminal mechanical injury. CyA also inhibited macrophage infiltration and neointimal formation in the wire-injured femoral arteries. These results suggested that calcineurin mediates vascular inflammation via stimulation of MCP-1 expression in VSMCs and macrophage infiltration. PMID- 14739160 TI - MCIP1 overexpression suppresses left ventricular remodeling and sustains cardiac function after myocardial infarction. AB - Pathological remodeling of the left ventricle (LV) after myocardial infarction (MI) is a major cause of heart failure. Although cardiac hypertrophy after increased loading conditions has been recognized as a clinical risk factor for human heart failure, it is unknown whether post-MI hypertrophic remodeling of the myocardium is beneficial for cardiac function over time, nor which regulatory pathways play a crucial role in this process. To address these questions, transgenic (TG) mice engineered to overexpress modulatory calcineurin-interacting protein-1 (MCIP1) in the myocardium were used to achieve cardiac-specific inhibition of calcineurin activation. MCIP1-TG mice and their wild-type (WT) littermates, were subjected to MI and analyzed 4 weeks later. At 4 weeks after MI, calcineurin was activated in the LV of WT mice, which was significantly reduced in MCIP1-TG mice. WT mice displayed a 78% increase in LV mass after MI, which was reduced by 38% in MCIP1-TG mice. Echocardiography indicated marked LV dilation and loss of systolic function in WT-MI mice, whereas TG-MI mice displayed a remarkable preservation of LV geometry and contractility, a pronounced reduction in myofiber hypertrophy, collagen deposition, and beta-MHC expression compared with WT-MI mice. Together, these results reveal a protective role for MCIP1 in the post-MI heart and suggest that calcineurin is a crucial regulator of postinfarction-induced pathological LV remodeling. The improvement in functional, structural, and molecular abnormalities in MCIP1-TG mice challenges the adaptive value of post-MI hypertrophy of the remote myocardium. The full text of this article is available online at http://circres.ahajournals.org. PMID- 14739161 TI - Transforming growth factor-beta-induced inhibition of myogenesis is mediated through Smad pathway and is modulated by microtubule dynamic stability. AB - The expression of muscle-specific genes associated with myogenesis is controlled by several myogenic transcription factors, including myogenin and MEF2D. Transforming growth factor-beta (TGF-beta) has been shown to inhibit myogenesis, yet the molecular mechanisms underlying such inhibition are not known. In the present study, TGF-beta was shown to inhibit myogenin and MEF2D expression and myotube formation in C2C12 myoblasts cultured in differentiation medium in a cell density-dependent manner. Transfection of C2C12 cells with Smad7, an antagonist for TGF-beta/Smad signaling, restored the capacity of these cells to differentiate in the presence of TGF-beta or when cultured in growth medium at low confluence, conditions that hinder muscle differentiation. Moreover, nocodazole, a microtubule-destabilizing agent, enhanced the inhibition of myogenesis exerted by TGF-beta, an effect that could be restored by tubulin polymerizing agent taxol, both of which have been shown to affect Smad microtubule interaction and regulate TGF-beta/Smad signaling. Our results indicate that TGF-beta inhibits myogenesis, at least in part, via Smad pathway, and provide evidence that low-dose pharmacological agents taxol and nocodazole can be used as a means to modulate myogenesis without affecting cell survival. PMID- 14739162 TI - Comparative evaluation of FGF-2-, VEGF-A-, and VEGF-C-induced angiogenesis, lymphangiogenesis, vascular fenestrations, and permeability. AB - Several endothelial growth factors induce both blood and lymphatic angiogenesis. However, a systematic comparative study of the impact of these factors on vascular morphology and function has been lacking. In this study, we report a quantitative analysis of the structure and macromolecular permeability of FGF-2-, VEGF-A-, and VEGF-C-induced blood and lymphatic vessels. Our results show that VEGF-A stimulated formation of disorganized, nascent vasculatures as a result of fusion of blood capillaries into premature plexuses with only a few lymphatic vessels. Ultrastructural analysis revealed that VEGF-A-induced blood vessels contained high numbers of endothelial fenestrations that mediated high permeability to ferritin, whereas the FGF-2-induced blood vessels lacked vascular fenestrations and showed only little leakage of ferritin. VEGF-C induced approximately equal amounts of blood and lymphatic capillaries with endothelial fenestrations present only on blood capillaries, mediating a medium level of ferritin leakage into the perivascular space. No endothelial fenestrations were found in FGF-2-, VEGF-A-, or VEGF-C-induced lymphatic vessels. These findings highlight the structural and functional differences between blood and lymphatic vessels induced by FGF-2, VEGF-A, and VEGF-C. Such information is important to consider in development of novel therapeutic strategies using these angiogenic factors. PMID- 14739163 TI - Marrow-derived stromal cells express genes encoding a broad spectrum of arteriogenic cytokines and promote in vitro and in vivo arteriogenesis through paracrine mechanisms. AB - We recently demonstrated that marrow stromal cells (MSCs) augment collateral remodeling through release of several cytokines such as VEGF and bFGF rather than via cell incorporation into new or remodeling vessels. The present study was designed to characterize the full spectrum of cytokine genes expressed by MSCs and to further examine the role of paracrine mechanisms that underpin their therapeutic potential. Normal human MSCs were cultured under normoxic or hypoxic conditions for 72 hours. The gene expression profile of the cells was determined using Affymetrix GeneChips representing 12 000 genes. A wide array of arteriogenic cytokine genes were expressed at baseline, and several were induced >1.5-fold by hypoxic stress. The gene array data were confirmed using ELISA assays and immunoblotting of the MSC conditioned media (MSC(CM)). MSC(CM) promoted in vitro proliferation and migration of endothelial cells in a dose dependent manner; anti-VEGF and anti-FGF antibodies only partially attenuated these effects. Similarly, MSC(CM) promoted smooth muscle cell proliferation and migration in a dose-dependent manner. Using a murine hindlimb ischemia model, murine MSC(CM) enhanced collateral flow recovery and remodeling, improved limb function, reduced the incidence of autoamputation, and attenuated muscle atrophy compared with control media. These data indicate that paracrine signaling is an important mediator of bone marrow cell therapy in tissue ischemia, and that cell incorporation into vessels is not a prerequisite for their effects. PMID- 14739164 TI - Chronic disease care. PMID- 14739165 TI - Reconfiguration of surgical, emergency, and trauma services in the United Kingdom. PMID- 14739166 TI - Treatment of acute pyelonephritis in children. PMID- 14739167 TI - Health claims for functional foods. PMID- 14739168 TI - Suspension of doctors. PMID- 14739170 TI - Parents convicted of killing to have their cases reviewed. PMID- 14739171 TI - Thousands of families to sue over retained organs. PMID- 14739172 TI - Claim of human reproductive cloning provokes calls for international ban. PMID- 14739173 TI - US government rejects WHO's attempts to improve diet. PMID- 14739176 TI - New infectious diseases will continue to emerge. PMID- 14739177 TI - Australia accused of blocking doctors visit to asylum seekers. PMID- 14739179 TI - Doctor accused of misrepresenting evidence in child abuse cases. PMID- 14739180 TI - Suspended consultant is reinstated. PMID- 14739181 TI - Whistleblower vows to fight on. PMID- 14739182 TI - Scientists beg EU to repeal new rules for clinical trials. PMID- 14739184 TI - MPs question effectiveness of Food Standards Agency on obesity. PMID- 14739185 TI - Drug company sues Spanish bulletin over fraud claim. PMID- 14739186 TI - Using an electrocautery strategy or recombinant follicle stimulating hormone to induce ovulation in polycystic ovary syndrome: randomised controlled trial. AB - OBJECTIVE: To compare the effectiveness of an electrocautery strategy with ovulation induction using recombinant follicle stimulating hormone in patients with polycystic ovary syndrome. DESIGN: Randomised controlled trial. SETTING: Secondary and tertiary hospitals in the Netherlands. PARTICIPANTS: 168 patients with clomiphene citrate resistant polycystic ovary syndrome: 83 were allocated electrocautery and 85 were allocated recombinant follicle stimulating hormone. INTERVENTION: Laparoscopic electrocautery of the ovaries followed by clomiphene citrate and recombinant follicle stimulating hormone if anovulation persisted, or induction of ovulation with recombinant follicle stimulating hormone. MAIN OUTCOME MEASURE: Ongoing pregnancy within 12 months. RESULTS: . The cumulative rate of ongoing pregnancy after recombinant follicle stimulating hormone was 67%. With only electrocautery it was 34%, which increased to 49% after clomiphene citrate was given. Subsequent recombinant follicle stimulating hormone increased the rate to 67% at 12 months (rate ratio 1.01, 95% confidence interval 0.81 to 1.24). No complications occurred from electrocautery with or without clomiphene citrate. Patients allocated to electrocautery had a significantly lower risk of multiple pregnancy (0.11, 0.01 to 0.86). CONCLUSION: The ongoing pregnancy rate from ovulation induction with laparoscopic electrocautery followed by clomiphene citrate and recombinant follicle stimulating hormone if anovulation persisted, or recombinant follicle stimulating hormone, seems equivalent to ovulation induction with recombinant follicle stimulating hormone, but the former procedure carries a lower risk of multiple pregnancy. PMID- 14739187 TI - Comparison of three methods for estimating rates of adverse events and rates of preventable adverse events in acute care hospitals. AB - OBJECTIVES: To compare the effectiveness, reliability, and acceptability of estimating rates of adverse events and rates of preventable adverse events using three methods: cross sectional (data gathered in one day), prospective (data gathered during hospital stay), and retrospective (review of medical records). DESIGN: Independent assessment of three methods applied to one sample. SETTING: 37 wards in seven hospitals (three public, four private) in southwestern France. PARTICIPANTS: 778 patients: medical (n = 278), surgical (n = 263), and obstetric (n = 237). MAIN OUTCOME MEASURES: The main outcome measures were the proportion of cases (patients with at least one adverse event) identified by each method compared with a reference list of cases confirmed by ward staff and the proportion of preventable cases (patients with at least one preventable adverse event). Secondary outcome measures were inter-rater reliability of screening and identification, perceived workload, and face validity of results. RESULTS: The prospective and retrospective methods identified similar numbers of medical and surgical cases (70% and 66% of the total, respectively) but the prospective method identified more preventable cases (64% and 40%, respectively), had good reliability for identification (kappa = 0.83), represented an acceptable workload, and had higher face validity. The cross sectional method showed a large number of false positives and identified none of the most serious adverse events. None of the methods was appropriate for obstetrics. CONCLUSION: The prospective method of data collection may be more appropriate for epidemiological studies that aim to convince clinical teams that their errors contribute significantly to adverse events, to study organisational and human factors, and to assess the impact of risk reduction programmes. PMID- 14739188 TI - Vaccinations against influenza and pneumococcus in children with diabetes: telephone questionnaire survey. PMID- 14739189 TI - Cohort study of examination performance of undergraduate medical students learning in community settings. AB - OBJECTIVES: To determine whether moving clinical medical education out of the tertiary hospital into a community setting compromises academic standards. DESIGN: Cohort study. SETTING: Flinders University four year graduate entry medical course. In their third year, students are able to choose to study at the tertiary teaching hospital in Adelaide, in rural general practices, or at Royal Darwin Hospital, a regional secondary referral hospital. PARTICIPANTS: All 371 medical students who did their year 3 study from 1998-2002. MAIN OUTCOME MEASURES: Mean student examination score (%) at the end of year 3. RESULTS: The unadjusted mean year 3 scores at each location differed significantly (P < 0.001); the mean score was 65.2 (SE = 0.43) for Adelaide students, 68.2 (0.83) for Darwin students, and 69.3 (0.97) for students on the rural programme. Mean year 2 scores were similar for each location. Post hoc tests of means adjusted for sex, age, year 2 score, and cohort year showed that the rural and Darwin groups had a significantly improved score in year 3 compared with the Adelaide group (adjusted mean difference = 3.08, 95% confidence interval 1.25 to 4.90, P < 0.001 for rural group; 1.91, 0.47 to 3.36, P = 0.001 for Darwin group). CONCLUSIONS: These findings show that the concern that student academic performance in the tertiary hospital would be better than that of students in the regional hospital and community settings is not justified. This challenges the orthodoxy of a tertiary hospital education being the gold standard for undergraduate medical students. PMID- 14739191 TI - Is folic acid the ultimate functional food component for disease prevention? PMID- 14739192 TI - Misinterpretation of serum cortisol in a patient with hyponatraemia. PMID- 14739193 TI - The problem of tobacco smoking. PMID- 14739194 TI - Rethinking management of chronic diseases. PMID- 14739195 TI - Can the NHS learn from US managed care organisations? PMID- 14739196 TI - Autism seems to be increasing worldwide, if not in London. PMID- 14739197 TI - Diagnosis of autism: adequate funding is needed for assessment services. PMID- 14739198 TI - Diagnosis of autism: use of autistic spectrum shows undisciplined thinking. PMID- 14739199 TI - Diagnosis of autism: current epidemic has social context. PMID- 14739200 TI - Children need international formulary to guarantee rational use of drugs. PMID- 14739201 TI - Guidelines on neuraminidase inhibitors in children are not supported by evidence. PMID- 14739202 TI - Prescription of heroin to treatment resistant heroin addicts: Dutch heroin trials show retention is better with methadone alone. PMID- 14739203 TI - Prescription of heroin to treatment resistant heroin addicts: double blinding is not possible. PMID- 14739204 TI - Prescription of heroin to treatment resistant heroin addicts: treatment needs to be multifaceted. PMID- 14739205 TI - Prescription of heroin to treatment resistant heroin addicts: replacement therapies need to be tested on a level playing field. PMID- 14739206 TI - Britain's failure to tackle research misconduct: Britain is ahead of most countries. PMID- 14739207 TI - Britain's failure to tackle research misconduct: UK bodies are now working together to set up panel on misconduct. PMID- 14739208 TI - Prescription of heroin to treatment resistant heroin addicts: heroin handouts are flawed policy. PMID- 14739209 TI - Editors and their priorities about libel and fraud: external checks must be imposed to protect the public. PMID- 14739210 TI - Editors and their priorities about libel and fraud: everyone in science must be active in its process. PMID- 14739211 TI - Editors and their priorities about libel and fraud: we need whistleblowers. PMID- 14739212 TI - Low-molecular-weight heparin versus low-dose aspirin in women with one fetal loss and a constitutional thrombophilic disorder. AB - The prospective evaluation of the effect of thromboprophylaxis in women with one unexplained pregnancy loss from the 10th week of amenorrhea was performed. A total of 160 patients with heterozygous factor V Leiden mutation, prothrombin G20210A mutation, or protein S deficiency were given 5 mg folic acid daily before conception, to be continued during pregnancy, and low-dose aspirin 100 mg daily or low-molecular-weight heparin enoxaparin 40 mg was taken from the 8th week. Twenty-three of the 80 patients treated with low-dose aspirin and 69 of the 80 patients treated with enoxaparin had a healthy live birth (odds ratio [OR], 15.5; 95% confidence interval [CI], 7-34, P <.0001). Enoxaparin was superior to low dose aspirin in each subgroup defined according to the underlying constitutional thrombophilic disorder. An associated protein Z deficiency and/or positive antiprotein Z antibodies were associated with poorer outcomes. The neonate weight was higher in the women successfully treated with enoxaparin, and neonates small for gestational age were more frequent in patients treated with low-dose aspirin. No significant side effects of the treatments could be evidenced in patients or newborns. As there is no argument to prove that low-dose aspirin may have been deleterious, these results support enoxaparin use during such at-risk pregnancies. PMID- 14739214 TI - PU.1 determines the self-renewal capacity of erythroid progenitor cells. AB - PU.1 is a hematopoietic-specific transcriptional activator that is absolutely required for the differentiation of B lymphocytes and myeloid-lineage cells. Although PU.1 is also expressed by early erythroid progenitor cells, its role in erythropoiesis, if any, is unknown. To investigate the relevance of PU.1 in erythropoiesis, we produced a line of PU.1-deficient mice carrying a green fluorescent protein reporter at this locus. We report here that PU.1 is tightly regulated during differentiation-it is expressed at low levels in erythroid progenitor cells and down-regulated upon terminal differentiation. Strikingly, PU.1-deficient fetal erythroid progenitors lose their self-renewal capacity and undergo proliferation arrest, premature differentiation, and apoptosis. In adult mice lacking one PU.1 allele, similar defects are detected following stress induced erythropoiesis. These studies identify PU.1 as a novel and critical regulator of erythropoiesis and highlight the versatility of this transcription factor in promoting or preventing differentiation depending on the hematopoietic lineage. PMID- 14739213 TI - Superior survival in primary systemic amyloidosis patients undergoing peripheral blood stem cell transplantation: a case-control study. AB - Primary systemic amyloidosis (AL) is a plasma cell dyscrasia resulting in multisystem failure and death. High-dose chemotherapy with peripheral blood stem cell transplantation (PBSCT) has been associated with higher response rates and seemingly higher overall survival than standard chemotherapy. Selection bias, however, confounds interpretation of these results. We performed a case-match control study comparing overall survival of 63 AL patients undergoing transplantation with 63 patients not undergoing transplantation. Matching criteria included age, sex, time to presentation, left ventricular ejection fraction, serum creatinine, septal thickness, nerve involvement, 24-hour urine protein, and serum alkaline phosphatase. According to design, there was no difference between the groups with respect to sex (57% males), age (median, 53 years), left ventricular ejection fraction (65%), number of patients with peripheral nerve involvement (17%), cardiac interventricular septal wall thickness (12 mm), serum creatinine (1.1 mg/dL [97.24 micromol/L]), and bone marrow plasmacytosis (8%). Sixty-six patients have died (16 cases and 50 controls). For PBSCT and control groups, respectively, the 1-, 2-, and 4-year overall survival rates are 89% and 71%; 81% and 55%; and 71% and 41%. Outside a randomized clinical trial, these results present the strongest data supporting the role of PBSCT in selected patients with AL. PMID- 14739215 TI - A fungal multicopper oxidase restores iron homeostasis in aceruloplasminemia. AB - Mutations that lead to a loss of the copper-containing plasma enzyme ceruloplasmin disrupt mammalian iron homeostasis. The mechanism by which ceruloplasmin mobilizes iron from cell stores has been controversial. We demonstrate that injection of a soluble copper-containing yeast protein Fet3p can restore iron homeostasis in phlebotomized mice with a deletion of the ceruloplasmin gene. These results show the conservation of function of copper containing proteins in eukaryotic iron metabolism. PMID- 14739216 TI - Hepatic low-density lipoprotein receptor-related protein deficiency in mice increases atherosclerosis independent of plasma cholesterol. AB - The low-density lipoprotein (LDL) receptor-related protein (LRP) has a well established role in the hepatic removal of atherogenic apolipoprotein E (APOE) rich remnant lipoproteins from plasma. In addition, LRP recognizes multiple distinct pro- and antiatherogenic ligands in vitro. Here, we investigated the role of hepatic LRP in atherogenesis independent of its role in removal of APOE rich remnant lipoproteins. Mice that allow inducible inactivation of hepatic LRP were combined with LDL receptor and APOE double-deficient mice (MX1Cre(+)LRP(flox/flox)LDLR(-/-)APOE(-/-)). On an LDLR(-/-)APOE(-/-) background, hepatic LRP deficiency resulted in decreased plasma cholesterol and triglycerides (cholesterol: 17.1 +/- 5.2 vs 23.4 +/- 6.3 mM, P =.025; triglycerides: 1.1 +/- 0.5 vs 2.2 +/- 0.8 mM, P =.002, for MX1Cre(+)LRP(flox/flox)-LDLR(-/-)APOE(-/-) and control LRP(flox/flox)-LDLR(-/-)APOE(-/-) mice, respectively). Lower plasma cholesterol in MX1Cre(+)LRP(flox/flox)-LDLR(-/-)APOE(-/-) mice coincided with increased plasma lipoprotein lipase (71.2 +/- 7.5 vs 19.1 +/- 2.4 ng/ml, P =.002), coagulation factor VIII (4.4 +/- 1.1 vs 1.9 +/- 0.5 U/mL, P =.001), von Willebrand factor (2.8 +/- 0.6 vs 1.4 +/- 0.3 U/mL, P =.001), and tissue-type plasminogen activator (1.7 +/- 0.7 vs 0.9 +/- 0.5 ng/ml, P =.008) compared with controls. Strikingly, MX1Cre(+)LRP(flox/flox)LDLR(-/-)APOE(-/-) mice showed a 2 fold higher atherosclerotic lesion area compared with controls (408.5 +/- 115.1 vs 219.1 +/- 86.0 10(3)microm(2), P =.003). Our data indicate that hepatic LRP plays a clear protective role in atherogenesis independent of plasma cholesterol, possibly due to maintaining low levels of its proatherogenic ligands. PMID- 14739218 TI - Autoimmune anemia in macaques following erythropoietin gene therapy. AB - We delivered the homologous erythropoietin (Epo) cDNA driven from a doxycycline regulated promoter via recombinant adeno-associated virus in skeletal muscle of 9 cynomolgus macaques. Upon induction, rapid supraphysiologic levels of Epo were obtained. Unexpectedly, some individuals developed a profound anemia that correlated with the appearance of neutralizing antibodies against the endogenous Epo. Both the endogenous erythropoietin and vector sequences were identical. This is the first example of the inadvertent development of an autoimmune disease in primates as a result of gene transfer of a gene expressing a self-antigen. It raises some concerns when a therapeutic protein is produced at high levels from an ectopic site. PMID- 14739217 TI - Rituximab and ICE as second-line therapy before autologous stem cell transplantation for relapsed or primary refractory diffuse large B-cell lymphoma. AB - Patients with relapsed or primary refractory diffuse large B-cell lymphoma (DLBCL) who achieve complete response (CR) before autologous stem cell transplantation (ASCT) generally have better outcomes than those who achieve only partial response (PR). We investigated whether adding rituximab to the ifosfamide carboplatin-etoposide (ICE) chemotherapy regimen (RICE) could increase the CR rate of patients with DLBCL under consideration for ASCT. Thirty-six eligible patients were treated with RICE, and 34 received all 3 planned cycles. The CR rate was 53%, significantly better than the 27% CR rate (P =.01) achieved among 147 similar consecutive historical control patients with DLBCL treated with ICE; the PR rate was 25%. Febrile neutropenia was the most frequent grade 3 or 4 nonhematologic toxicity; it occurred in 7.5% of delivered cycles. No patient had RICE-related toxicity that precluded ASCT. The median number of CD34(+) cells per kilogram mobilized was 6.3 x 10(6). Progression-free survival rates of patients who underwent transplantation after RICE were marginally better than those of 95 consecutive historical control patients who underwent transplantation after ICE (54% vs 43% at 2 years; P =.25). RICE appears to induce very high CR rates in patients with relapsed and refractory DLBCL; however, further studies are necessary to determine whether this treatment regimen will improve outcomes after ASCT. PMID- 14739219 TI - Retraction: The triterpenoid CDDO-Imidazolide induces apoptosis and enhances fludarabine-induced apoptosis of CLL B-cells. PMID- 14739221 TI - Induction of human T-cell tolerance to porcine xenoantigens through mixed hematopoietic chimerism. AB - Xenotransplantation from pigs could provide a potential solution to the severe shortage of allogeneic donor organs. Because xenogeneic tissues are subject to vigorous immune rejection, tolerance induction is likely to be essential to the success of clinical xenotransplantation. Here we explore the possibility of inducing human T-cell tolerance to porcine xenografts through mixed chimerism. We previously showed that NOD/SCID-Tg mice expressing porcine cytokine transgenes permit the induction of durable porcine hematopoietic chimerism. In this study we achieved human T-cell development in these mice by engrafting human fetal thymus/liver tissues. In porcine hematopoietic chimeras, human thymus grafts were populated with porcine class II(high) cells in addition to human cells, and human T cells were tolerant of the porcine hematopoietic donor as measured by mixed lymphocyte reaction assay and skin grafting. This study proves the principle that porcine chimerism induces tolerance of xenoreactive human T cells. PMID- 14739220 TI - Genetic variation responsible for mouse strain differences in integrin alpha 2 expression is associated with altered platelet responses to collagen. AB - As mouse models have become commonplace for studying hemostasis and thrombosis, we considered whether the mouse system had utility for assessing genetic alterations in platelet receptors. Platelets from 5 mouse strains (C57BL/6 [C57], FVB/N [FVB], BALB/c, C3H/He, and 129Sv) showed only minor differences in the expression of integrin alpha(IIb), integrin beta(3), glycoprotein (GP) Ib alpha, or GPVI across strains. However, FVB platelets expressed approximately 50% the level of integrin alpha(2) as platelets from other strains (P <.0001). We bred FVB mice with C57 and assessed alpha(2) expression in FVB/C57xFVB/C57 (F2) offspring. Linkage analysis demonstrated the gene responsible for alpha(2) levels is tightly linked to the D13mit260 marker (log odds [lod] score 6.7) near the alpha(2) gene. FVB platelets showed reduced aggregation and a longer lag phase to collagen. FVB and C57 platelets aggregated similarly to collagen-related peptide, but FVB platelets showed a reduction in rhodocytin-induced Syk and PLC gamma 2 tyrosine phosphorylation. Thus, FVB platelets express half the level of alpha(2) as other mouse strains, a trait linked to the alpha(2) gene and seemingly responsible for reduced platelet aggregation to collagen. These strain differences serve as a useful model for the 2-fold difference in human platelet alpha(2)beta(1) expression and demonstrate that alpha(2)beta(1) participates in signaling during platelet activation. PMID- 14739222 TI - Atypical features of familial hemophagocytic lymphohistiocytosis. AB - Familial hemophagocytic lymphohistiocytosis (FHLH) is a rare, rapidly progressive disorder of early childhood characterized by uncontrolled activation of T cells and macrophages. Although perforin gene mutations have been described in a proportion of patients with FHLH, the genotype/phenotype correlation is still limited. Only a few patients with late onset clinical manifestations have been reported. The biochemical and immunologic alterations in the asymptomatic phase are not well known. We report on a family in which 2 fraternal twins both homozygous for a perforin mutation previously described as causative of the disease, markedly differed in phenotypic expression of FHLH. The twins also had a second novel heterozygous mutation. Natural killer (NK) activity was severely impaired in the patient and was normal in the asymptomatic fraternal twin. Our report highlights that FHLH may present after a long disease-free interval during which biochemical or immunologic alterations may be not evident, thus implying a role for interfering factors. PMID- 14739223 TI - Thrombin-activatable fibrinolysis inhibitor and the risk for recurrent venous thromboembolism. AB - The impact of fibrinolysis for predicting the risk for recurrent venous thromboembolism (VTE) is low. We prospectively followed up 600 patients with a first VTE and evaluated the thrombin-activatable fibrinolysis inhibitor (TAFI) as a risk factor for recurrence. A high TAFI level (75th or higher percentile in thrombosis patients) was associated with a 2-fold higher risk for recurrence compared with lower levels. The probability of recurrence 2 years after anticoagulation was 14.5% (95% confidence interval [CI], 8.6-20.4) among patients with high TAFI levels and 6.8% (95% CI, 4.3-9.3) among patients with lower levels (P =.006). Our data also support the concept of a linkage between fibrinolysis and the coagulation system. Patients with high TAFI levels had significantly higher levels of factors XI, VIII, and IX, and a high risk of recurrence was seen among patients with high TAFI levels and high levels of one of these factors. The relative risk (RR) for recurrence was highest among patients with high TAFI and high factor XI (RR, 2.9; 95% CI, 1.3-6.9), high factor VIII (RR, 6.5; 95% CI, 2.9 14.8), or high factor IX (RR, 2.0; 95% CI, 1.0-3.9) levels compared with patients with low levels of TAFI and one of these factors. PMID- 14739224 TI - Gamma-catenin contributes to leukemogenesis induced by AML-associated translocation products by increasing the self-renewal of very primitive progenitor cells. AB - Acute myeloid leukemia (AML) is characterized by the block of differentiation, deregulated apoptosis, and an increased self-renewal of hematopoietic precursors. It is unclear whether the self-renewal of leukemic blasts results from the cumulative effects of blocked differentiation and impaired apoptosis or whether there are mechanisms directly increasing self-renewal. The AML-associated translocation products (AATPs) promyelocytic leukemia/retinoic acid receptor alpha (PML/RAR alpha), promyelocytic leukemia zinc finger (PLZF)/RAR alpha (X-RAR alpha), and AML-1/ETO block hematopoietic differentiation. The AATPs activate the Wnt signaling by up-regulating gamma-catenin. Activation of the Wnt signaling augments self-renewal of hematopoietic stem cells (HSCs). Therefore, we investigated how AATPs influence self-renewal of HSCs and evaluated the role of gamma-catenin in the determination of the phenotype of HSCs expressing AATPs. Here we show that the AATPs directly activate the gamma-catenin promoter. The crucial role of gamma-catenin in increasing the self-renewal of HSCs upon expression of AATPs is demonstrated by (i) the abrogation of replating efficiency upon hindrance of gamma-catenin expression through RNA interference, and (ii) the augmentation of replating efficiency of HSCs upon overexpression of gamma-catenin itself. In addition, the inoculation of gamma-catenin-transduced HSCs into irradiated recipient mice establishes the clinical picture of AML. These data provide the first evidence that the aberrant activation of Wnt signaling by the AATP decisively contributes to the pathogenesis of AML. PMID- 14739225 TI - Human immunodeficiency virus facilitates infection/replication of hepatitis C virus in native human macrophages. AB - Hepatitis C virus (HCV) was found to replicate in monocytes/macrophages particularly in patients with human immunodeficiency virus type 1 (HIV-1) infection. This study was undertaken to determine whether HIV facilitates HCV infection of native human macrophages in vitro. Monocytes/macrophages were collected from healthy donors, infected with HIV M-tropic molecular clone, and then exposed to HCV-positive sera. Presence of positive and negative HCV RNA strands was determined with a novel strand-specific quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR). Preceding as well as near-simultaneous infection with HIV made the macrophages more susceptible to infection with HCV; in particular, an HCV RNA-negative strand was detectable almost exclusively in the setting of concomitant HIV infection. Furthermore, HCV RNA load correlated with HIV replication level in the early stage of infection. The ratio of positive to negative strand in macrophages was lower than in control liver samples. HIV infection was also found to facilitate HCV replication in a Daudi B-cell line with engineered CD4 expression. It seems that HIV infection can facilitate replication of HCV in monocytes/macrophages either by rendering cells more susceptible to HCV infection or by increasing HCV replication. This could explain the presence of extrahepatic HCV replication in HIV-coinfected individuals. PMID- 14739226 TI - Inappropriately low reticulocytosis in severe malarial anemia correlates with suppression in the development of late erythroid precursors. AB - Inappropriately low reticulocytosis may exacerbate malarial anemia, but the under lying mechanism is not clear. In this study, naive and infected mice were treated with recombinant murine erythropoietin (EPO), and the upstream events of erythropoiesis affected by blood-stage Plasmodium chabaudi AS were investigated. Malaria infection, with or without EPO treatment, led to a suboptimal increase in TER119(+) erythroblasts compared with EPO-treated naive mice. Furthermore, a lower percentage of TER119(+) erythroblasts in infected mice were undergoing terminal differentiation to become mature hemoglobin-producing erythroblasts. The impaired maturation of erythroblasts during infection was associated with a shift in the transferrin receptor (CD71) expression from the TER119(+) population to B220(+) population. Moreover, the suboptimal increase in TER119(+) erythroblasts during infection coincided with a blunted proliferative response by splenocytes to EPO stimulation in vitro, although a high frequency of these splenocytes expressed EPO receptor (EPOR). Taken together, these data suggest that during malaria, EPO-induced proliferation of early EPOR-positive erythroid progenitors is suppressed, which may lead to a suboptimal generation of TER119(+) erythroblasts. The shift in CD71 expression may result in impaired terminal maturation of these erythroblasts. Thus, inadequate reticulocytosis during malaria is associated with suppressed proliferation, differentiation, and maturation of erythroid precursors. PMID- 14739227 TI - Efficient lentiviral gene transfer to canine repopulating cells using an overnight transduction protocol. AB - The use of lentiviral vectors for the transduction of hematopoietic stem cells has evoked much interest owing to their ability to stably integrate into the genome of nondividing cells. However, published large animal studies have reported highly variable gene transfer rates of typically less than 1%. Here we report the use of lentiviral vectors for the transduction of canine CD34(+) hematopoietic repopulating cells using a very short, 18-hour transduction protocol. We compared lentiviral transduction of hematopoietic repopulating cells from either stem cell factor (SCF)- and granulocyte-colony stimulating factor (G CSF)-primed marrow or mobilized peripheral blood in a competitive repopulation assay in 3 dogs. All dogs engrafted rapidly within 9 days. Transgene expression was detected in all lineages (B cells, T cells, granulocytes, and red blood cells as well as platelets) indicating multilineage engraftment of transduced cells, with overall long-term marking levels of up to 12%. Gene transfer levels in mobilized peripheral blood cells were slightly higher than in primed marrow cells. In conclusion, we show efficient lentiviral transduction of canine repopulating cells using an overnight transduction protocol. These results have important implications for the design of stem cell gene therapy protocols, especially for those diseases in which the maintenance of stem cells in culture is a major limitation. PMID- 14739228 TI - Identification and characterization of a very low density lipoprotein receptor binding peptide from tissue factor pathway inhibitor that has antitumor and antiangiogenic activity. AB - Tissue factor pathway inhibitor (TFPI) is the major physiologic inhibitor of the extrinsic coagulation pathway. We have previously shown that TFPI is also a potent inhibitor of endothelial proliferation in vitro and of primary and metastatic tumor growth in vivo. Surprisingly, the antitumor activity of TFPI was demonstrated to be independent of its anticoagulant activity, suggesting a possible nonhemostatic mechanism of action for TFPI in these models. This antitumor mechanism may involve the very low density lipoprotein (VLDL) receptor because the in vitro antiproliferative activity of TFPI is mediated through interaction with the VLDL receptor. In the current study, we identify a 23-amino acid fragment of TFPI (TFPIc23) localized to the C-terminus, which mediates binding to the VLDL receptor. The TFPIc23 peptide inhibits endothelial cell proliferation through an apoptotic mechanism and blocks vessel outgrowth in the in vitro assays, and this activity is mediated through interaction with the VLDL receptor. In vivo, this peptide potently inhibits angiogenesis in Matrigel and chick chorioallantoic membrane models and also inhibits metastatic tumor growth. Our data demonstrate that this VLDL receptor-binding fragment of the TFPI molecule has apoptotic, antiangiogenic, and antitumor activity and suggests a possible mechanism whereby TFPI can regulate angiogenesis and tumor growth independently of its anticoagulant activity. PMID- 14739229 TI - The granzyme B-serglycin complex from cytotoxic granules requires dynamin for endocytosis. AB - Cytotoxic T lymphocytes and natural killer cells destroy target cells via the directed exocytosis of lytic effector molecules such as perforin and granzymes. The mechanism by which these proteins enter targets is uncertain. There is ongoing debate over whether the most important endocytic mechanism is nonspecific or is dependent on the cation-independent mannose 6-phosphate receptor. This study tested whether granzyme B endocytosis is facilitated by dynamin, a key factor in many endocytic pathways. Uptake of and killing by the purified granzyme B molecule occurred by both dynamin-dependent and -independent mechanisms. However most importantly, serglycin-bound granzyme B in high-molecular-weight degranulate material from cytotoxic T lymphocytes predominantly followed a dynamin-dependent pathway to kill target cells. Similarly, killing by live cytotoxic T lymphocytes was attenuated by a defect in the dynamin endocytic pathway, and in particular, the pathways characteristically activated by granzyme B were affected. We therefore propose a model where degranulated serglycin-bound granzymes require dynamin for uptake. PMID- 14739230 TI - Pericapillary hemorrhage as criterion of severe human digestive graft-versus-host disease. AB - In an experimental model we demonstrated that endothelial cells of all organs are targets of the alloimmune reaction. Here, in 68 digestive biopsies, we found endothelial lesions by immunohistochemistry and ultrastructure in patients with severe acute graft-versus-host disease (GVHD). In contrast, no such endothelial cell alterations were found either in patients without GVHD or in nongrafted controls. In the biopsies with severe GVHD lesions, ultrastructure showed rupture of the capillary basal membrane and extravased red blood cells. These pericapillary hemorrhages were highly correlated with GVHD severity. In a separate cohort of 39 patients who underwent an allogeneic transplantation after a nonmyeloablative conditioning, 8 patients had intestinal biopsies. Three of these latter patients had both severe pathologic lesions of GVHD and similar endothelial lesions, thus, strengthening the concept that endothelial lesions are linked to GVHD severity and not to the intensity of the conditioning regimen. (Blood. 2004;103:4681-4684) PMID- 14739231 TI - A novel approach for evaluating the efficiency of siRNAs on protein levels in cultured cells. AB - An important aim in the post-sequencing age of functional genomics is to translate gene sequences into protein functions. This shift of focus is particularly necessary for a very large number of human genes, referred to as novel genes, where we have no or very rudimentary information about their biochemical functions. Recently, a new method for investigating human gene functions using small interfering RNA molecules (siRNAs) has become available. siRNAs are powerful reagents for post-transcriptional silencing, where mRNA targeted by the siRNAs is degraded in vivo and the level of the encoded protein is reduced. However, the lack of antibodies against proteins encoded by novel genes restricts the general value of siRNAs as functional tools, as only the mRNA levels can be measured for these genes. We report a method that combines measurements of protein levels in cell culture for novel, exogenously expressed genes, with parallel measurements of the endogenous mRNA levels of the same genes. We find that this combinatorial approach correctly predicts siRNAs that efficiently reduce mRNA and protein levels in cultured cells. Furthermore, this method identifies proteins that have a slow turnover, which weakens the value of the RNA interference method as a tool for functional studies of such genes. The described method should prove to be valuable for large-scale functional studies of novel human genes. PMID- 14739232 TI - Construction of long DNA molecules using long PCR-based fusion of several fragments simultaneously. AB - A procedure for precise assembly of linear DNA constructs as long as 20 kb is proposed. The method, which we call long multiple fusion, has been used to assemble up to four fragments simultaneously (for a 10.8 kb final product), offering an additional improvement on the combination of long PCR and overlap extension PCR. The method is based on Pfu polymerase mix, which has a proofreading activity. We successfully assembled (and confirmed by sequencing) seven different linear constructs ranging from 3 to 20 kb, including two 20 kb products (from fragments of 11, 1.7 and 7.5 kb), two 10.8 kb constructs, and two constructs of 6.1 and 6.2 kb, respectively. Accuracy of the PCR fusion is greater than or equal to one error per 6.6 kb, which is consistent with the expected error rate of the PCR mix. The method is expected to facilitate various kinds of complex genetic engineering projects that require precise in-frame assembly of multiple fragments, such as somatic cell knockout in human cells or creation of whole genomes of viruses for vaccine research. PMID- 14739233 TI - Impedance-based detection of DNA sequences using a silicon transducer with PNA as the probe layer. AB - Electrochemical impedance measurements were used for the detection of single strand DNA sequences using a peptide nucleic acid (PNA) probe layer immobilized onto Si/SiO2 chips. An epoxysilane layer is first immobilized onto the Si/SiO2 surface. The immobilization procedure consists of an epoxide/amine coupling reaction between the amino group of the PNA linker and the epoxide group of the silane. A 20-nucleotide sequence of PNA was used. Impedance measurements allow for the detection of the changes in charge distribution at the oxide/solution interface following modifications to the oxide surface. Due to these modifications, there are significant shifts in the semiconductor's flat-band potential after immobilization and hybridization. The results obtained using this direct and rapid approach are supported by fluorescence measurements according to classical methods for the detection of nucleic acid sequences. PMID- 14739234 TI - Specific inhibitors of HCV polymerase identified using an NS5B with lower affinity for template/primer substrate. AB - The interaction of the hepatitis C virus (HCV) RNA-dependent RNA polymerase with RNA substrate is incompletely defined. We have characterized the activities of the HCV NS5B polymerase, modified by different deletions and affinity tags, with a routinely used homopolymeric substrate, and established apparent affinities of the various NS5B constructs both for the NTP and the template/primer substrates. We identified a uniquely tagged HCV NS5B RNA polymerase construct with a lower affinity (higher K(m)) than mature HCV NS5B for template/ primer substrate and highlighted the use of such a polymerase for the identification of inhibitors of NS5B activity, particularly inhibitors of productive RNA binding. The characterization of specific benzimidazole-5-carboxamide-based inhibitors, identified in a screening campaign, revealed that this class of compounds was non competitive with regard to NTP incorporation and had no effect on processive elongation, but inhibited an initiation phase of the HCV polymerase activity. The potency of these compounds versus a panel of different NS5B polymerase constructs was inversely proportional to the enzymes' affinities for template/primer substrate. The benzimidazole-5-carboxamide compounds also inhibited the full length, untagged NS5B de novo initiation reaction using HCV 3'-UTR substrate RNA and expand the diversifying pool of potential HCV replication inhibitors. PMID- 14739235 TI - Formation of an intramolecular triple-stranded DNA structure monitored by fluorescence of 2-aminopurine or 6-methylisoxanthopterin. AB - The parallel (recombination) 'R-triplex' can accommodate any nucleotide sequence with the two identical DNA strands in parallel orientation. We have studied oligonucleotides able to fold back into such a recombination-like structure. We show that the fluorescent base analogs 2-aminopurine (2AP) and 6 methylisoxanthopterin (6MI) can be used as structural probes for monitoring the integrity of the triple-stranded conformation and for deriving the thermodynamic characteristics of these structures. A single adenine or guanine base in the third strand of the triplex-forming and the control oligonucleotides, as well as in the double-stranded (ds) and single-stranded (ss) reference molecules, was substituted with 2AP or 6MI. The 2AP*(T.A) and 6MI*(C.G) triplets were monitored by their fluorescence emission and the thermal denaturation curves were analyzed with a quasi-two-state model. The fluorescence of 2AP introduced into an oligonucleotide sequence unable to form a triplex served as a negative control. We observed a remarkable similarity between the thermodynamic parameters derived from melting of the secondary structures monitored through absorption of all bases at 260 nm or from fluorescence of the single base analog. The similarity suggests that fluorescence of the 2AP and 6MI base analogs may be used to monitor the structural disposition of the third strand. We consider the data in the light of alternative 'branch migration' and 'strand exchange' structures and discuss why these are less likely than the R-type triplex. PMID- 14739236 TI - Exon repetition: a major pathway for processing mRNA of some genes is allele specific. AB - Exon repetition describes the presence of tandemly repeated exons in mRNA in the absence of duplications in the genome. Its existence challenges our understanding of gene expression, because the linear organization of sequences in apparently normal genes must be subverted during RNA synthesis or processing. It is restricted to a small number of genes in some of which over half of the mRNA contains specific patterns of repetition. Although it is sometimes assumed to arise by trans-splicing, there is no evidence of this and the efficiency is very much higher than for examples of bona fide trans-splicing in mammals. Furthermore, a potentially ubiquitous reaction such as trans-splicing is not consistent with a phenomenon that involves such a high proportion of the products of so few genes. Instead, it seems more probable that exon repetition is caused by a specific trans-acting factor. We have tested this and demonstrate for the two best characterized examples that the property is restricted to specific alleles of the affected genes and is determined in cis. It is not determined by exonic splicing signals, as had been suggested previously. In heterozygotes, RNA transcribed from the two alleles of an affected gene can have fundamentally different fates. PMID- 14739237 TI - Statistical resynchronization and Bayesian detection of periodically expressed genes. AB - We propose a periodic-normal mixture (PNM) model to fit transcription profiles of periodically expressed (PE) genes in cell cycle microarray experiments. The model leads to a principled statistical estimation procedure that produces more accurate estimates of the mean cell cycle length and the gene expression periodicity than existing heuristic approaches. A central component of the proposed procedure is the resynchronization of the observed transcription profile of each PE gene according to the PNM with estimated periodicity parameters. By using a two-component mixture-Beta model to approximate the PNM fitting residuals, we employ an empirical Bayes method to detect PE genes. We estimate that about one-third of the genes in the genome of Saccharomyces cerevisiae are likely to be transcribed periodically, and identify 822 genes whose posterior probabilities of being PE are greater than 0.95. Among these 822 genes, 540 are also in the list of 800 genes detected by Spellman. Gene ontology annotation analysis shows that many of the 822 genes were involved in important cell cycle related processes, functions and components. When matching the 822 resynchronized expression profiles of three independent experiments, little phase shifts were observed, indicating that the three synchronization methods might have brought cells to the same phase at the time of release. PMID- 14739238 TI - Crystal structure of the catalytic fragment of murine poly(ADP-ribose) polymerase 2. AB - Poly(ADP-ribose) polymerase-1 (PARP-1) has become an important pharmacological target in the treatment of cancer due to its cellular role as a 'DNA-strand break sensor', which leads in part to resistance to some existing chemo- and radiological treatments. Inhibitors have now been developed which prevent PARP-1 from synthesizing poly(ADP-ribose) in response to DNA-breaks and potentiate the cytotoxicity of DNA damaging agents. However, with the recent discoveries of PARP 2, which has a similar DNA-damage dependent catalytic activity, and additional members containing the 'PARP catalytic' signature, the isoform selectivity and resultant pharmacological effects of existing inhibitors are brought into question. We present here the crystal structure of the catalytic fragment of murine PARP-2, at 2.8 A resolution, and compare this to the catalytic fragment of PARP-1, with an emphasis on providing a possible framework for rational drug design in order to develop future isoform-specific inhibitors. PMID- 14739240 TI - Four new avian mitochondrial genomes help get to basic evolutionary questions in the late cretaceous. AB - Good phylogenetic trees are required to test hypotheses about evolutionary processes. We report four new avian mitochondrial genomes, which together with an improved method of phylogenetic analysis for vertebrate mt genomes give results for three questions in avian evolution. The new mt genomes are: magpie goose (Anseranas semipalmata), an owl (morepork, Ninox novaeseelandiae); a basal passerine (rifleman, or New Zealand wren, Acanthisitta chloris); and a parrot (kakapo or owl-parrot, Strigops habroptilus). The magpie goose provides an important new calibration point for avian evolution because the well-studied Presbyornis fossils are on the lineage to ducks and geese, after the separation of the magpie goose. We find, as with other animal mitochondrial genomes, that RY coding is helpful in adjusting for biases between pyrimidines and between purines. When RY-coding is used at third positions of the codon, the root occurs between paleognath and neognath birds (as expected from morphological and nuclear data). In addition, passerines form a relatively old group in Neoaves, and many modern avian lineages diverged during the Cretaceous. Although many aspects of the avian tree are stable, additional taxon sampling is required. PMID- 14739239 TI - A primordial RNA modification enzyme: the case of tRNA (m1A) methyltransferase. AB - The modified nucleoside 1-methyladenosine (m(1)A) is found in the T-loop of many tRNAs from organisms belonging to the three domains of life (Eukaryota, Bacteria, Archaea). In the T-loop of eukaryotic and bacterial tRNAs, m(1)A is present at position 58, whereas in archaeal tRNAs it is present at position(s) 58 and/or 57, m(1)A57 being the obligatory intermediate in the biosynthesis of 1-methylinosine (m(1)I57). In yeast, the formation of m(1)A58 is catalysed by the essential tRNA (m(1)A58) methyltransferase (MTase), a tetrameric enzyme that is composed of two types of subunits (Gcd14p and Gcd10p), whereas in the bacterium Thermus thermophilus the enzyme is a homotetramer of the TrmI polypeptide. Here, we report that the TrmI enzyme from the archaeon Pyrococcus abyssi is also a homotetramer. However, unlike the bacterial site-specific TrmI MTase, the P.abyssi enzyme is region-specific and catalyses the formation of m(1)A at two adjacent positions (57 and 58) in the T-loop of certain tRNAs. The stabilisation of P.abyssi TrmI at extreme temperatures involves intersubunit disulphide bridges that reinforce the tetrameric oligomerisation, as revealed by biochemical and crystallographic evidences. The origin and evolution of m(1)A MTases is discussed in the context of different hypotheses of the tree of life. PMID- 14739241 TI - Maternal and paternal lineages in cross-breeding bovine species. Has wisent a hybrid origin? AB - The tribe Bovini comprises cattle and cattle-like species. Reconstructions of their phylogeny have so far been incomplete and have yielded conflicting conclusions about the relationship of American bison and wisent (European bison). We have compared the sequences of three mitochondrial and two Y-chromosomal DNA segments. Mitochondrial DNA indicates that four distinct maternal lineages diverged after an early split-off of the buffalo species, leading to (1) taurine cattle and zebu, (2) wisent, (3) American bison and yak, and (4) banteng, gaur, and gayal, respectively. At a higher level, lineages (1) and (2) and lineages (3) and (4) are probably associated. In contrast, Y-chromosomal sequences indicate a close association of American and European bison, which is in agreement with their morphological similarity, complete fertility of hybrid offspring, and amplified fragment length polymorphism (AFLP) fingerprints of nuclear DNA. One explanation for the anomalous divergence of the mitochondrial DNA from the two bison species is lineage sorting, which implies that two distinct mitochondrial lineages coexisted in the bison-yak branch until the recent divergence of American bison and wisent. Alternatively, the wisent may have emerged by species hybridization initiated by introgression of bison bulls in another ancestral species. This "transpatric" mode of species formation would be consistent with the recent appearance of the wisent in the fossil record without clearly identifiable ancestors. PMID- 14739242 TI - Endosymbiont phylogenesis in the dryophthoridae weevils: evidence for bacterial replacement. AB - Intracellular symbiosis is widespread in the insect world where it plays an important role in evolution and adaptation. The weevil family Dryophthoridae (Curculionoidea) is of particular interest in intracellular symbiosis evolution with regard to the great economical and ecological features of these invasive insects, and the potential for comparative studies across a wide range of host plants and environments. Here, we have analyzed the intracellular symbiotic bacteria of 19 Dryophthoridae species collected worldwide, representing a wide range of plant species and tissues. All except one (Sitophilus linearis) harbor symbiotic bacteria within specialized cells (the bacteriocytes) assembled as an organ, the bacteriome. Phylogenetic analysis of the 16S rDNA gene sequence of the Dryophthoridae endosymbionts revealed three endosymbiotic clades belonging to gamma3-Proteobacteria and characterized by different GC contents and evolutionary rate. The genus name Candidatus Nardonella was proposed for the ancestral clade infesting Dryophthoridae 100 MYA and represented by five of nine bacterial genera studied. For this clade showing low GC content (40.5% GC) and high evolutionary rate (0.128 substitutions/site per 100 Myr), a single infection and subsequent cospeciation of the host and the endosymbionts was observed. In the two other insect lineage endosymbionts, with relatively high GC content (53.4% and 53.8% GC), competition with ancestral pathogenic bacteria might have occurred, leading to endosymbiont replacement in present-day last insects. PMID- 14739243 TI - Recent intron gain in elongation factor-1alpha of colletid bees (Hymenoptera: Colletidae). AB - We discovered the presence of a unique spliceosomal intron in the F1 copy of elongation factor-1alpha (EF-1alpha) restricted to the bee family Colletidae (Hymenoptera: Apoidae). The intron ranges in size from 101 to 1044 bp and shows no positional sliding. Our data also demonstrate the complete absence of this intron from exemplars representing all other bee families, as well as from close hymenopteran relatives. A review of the literature finds that this intron is likewise absent from all other arthropods for which data are available. This provides unambiguous evidence for a relatively recent intron insertion event in the colletid common ancestor and, at least in this specific instance, lends support to the introns-late hypothesis. The comparative distribution of this novel intron also supports the monophyly of Colletidae and the exclusion of the Stenotritidae from this family, providing an example of the potential of some introns to act as robust markers of shared descent. PMID- 14739244 TI - Deriving the genomic tree of life in the presence of horizontal gene transfer: conditioned reconstruction. AB - The horizontal gene transfer (HGT) being inferred within prokaryotic genomes appears to be sufficiently massive that many scientists think it may have effectively obscured much of the history of life recorded in DNA. Here, we demonstrate that the tree of life can be reconstructed even in the presence of extensive HGT, provided the processes of genome evolution are properly modeled. We show that the dynamic deletions and insertions of genes that occur during genome evolution, including those introduced by HGT, may be modeled using techniques similar to those used to model nucleotide substitutions that occur during sequence evolution. In particular, we show that appropriately designed general Markov models are reasonable tools for reconstructing genome evolution. These studies indicate that, provided genomes contain sufficiently many genes and that the Markov assumptions are met, it is possible to reconstruct the tree of life. We also consider the fusion of genomes, a process not encountered in gene sequence evolution, and derive a method for the identification and reconstruction of genome fusion events. Genomic reconstructions of a well-defined classical four genome problem, the root of the multicellular animals, show that the method, when used in conjunction with paralinear/logdet distances, performs remarkably well and is relatively unaffected by the recently discovered big genome artifact. PMID- 14739245 TI - Rapid sequence turnover at an intergenic locus in Drosophila. AB - Closely related species of Drosophila tend to have similar genome sizes. The strong imbalance in favor of small deletions relative to insertions implies that the unconstrained DNA in Drosophila is unlikely to be passively inherited from even closely related ancestors, and yet most DNA in Drosophila genomes is intergenic and potentially unconstrained. In an attempt to investigate the maintenance of this intergenic DNA, we studied the evolution of an intergenic locus on the fourth chromosome of the Drosophila melanogaster genome. This 1.2-kb locus is marked by two distinct, large insertion events: a nuclear transposition of a mitochondrial sequence and a transposition of a nonautonomous DNA transposon DNAREP1_DM. Because we could trace the evolutionary histories of these sequences, we were able to reconstruct the length evolution of this region in some detail. We sequenced this locus in all four species of the D. melanogaster species complex: D. melanogaster, D. simulans, D. sechellia, and D. mauritiana. Although this locus is similar in size in these four species, less than 10% of the sequence from the most recent common ancestor remains in D. melanogaster and all of its sister species. This region appears to have increased in size through several distinct insertions in the ancestor of the D. melanogaster species complex and has been shrinking since the split of these lineages. In addition, we found no evidence suggesting that the size of this locus has been maintained over evolutionary time; these results are consistent with the model of a dynamic equilibrium between persistent DNA loss through small deletions and more sporadic DNA gain through less frequent but longer insertions. The apparent stability of genome size in Drosophila may belie very rapid sequence turnover at intergenic loci. PMID- 14739246 TI - Rapid evolution of a pollen-specific oleosin-like gene family from Arabidopsis thaliana and closely related species. AB - It has been shown in a variety of species that genes expressed in reproductive tissues evolve rapidly, which often appears to be the result of positive Darwinian selection. We investigated the evolution of a family of seven pollen specific oleosin-like proteins (or oleopollenins) in Arabidopsis thaliana and two closely related species. More than 30 kb of a genomic region that harbors the complete, tandemly repeated oleopollenin cluster were sequenced from Arabidopsis lyrata ssp. lyrata, and Boechera drummondii. A phylogenetic analysis of the complete gene cluster from these three species and from Brassica oleracea confirmed its rapid evolution resulting from gene duplication and gene loss events, numerous amino acid substitutions, and insertions/deletions in the coding sequence. Independent duplications were inferred in the lineages leading to Arabidopsis and to Brassica, and gene loss was inferred in the lineage leading to B. drummondii. Comparisons of the ratio of nonsynonymous (d(N)) and synonymous (d(S)) divergence revealed that the oleopollenins are among the most rapidly evolving proteins currently known from Arabidopsis and that they may evolve under positive Darwinian selection. Reverse transcriptase polymerase chain reaction analysis demonstrated the expression of oleopollenins in flowers of the outcrossing A. lyrata, the selfing B. drummondii, and the apomictic Boechera holboellii, suggesting that oleopollenins play an important role in species with different breeding systems. These results are consistent with a putative function in species recognition, but further analyses of protein function and sequence variation in species with different breeding systems are necessary to reveal the underlying causes for the rapid evolution of oleopollenins. PMID- 14739247 TI - Bacterial proteins predisposed for targeting to mitochondria. AB - Mitochondria evolved from an endosymbiotic proteobacterium in a process that required the transfer of genes from the bacterium to the host cell nucleus, and the translocation of proteins thereby made in the host cell cytosol into the internal compartments of the organelle. According to current models for this evolution, two highly improbable events are required to occur simultaneously: creation of a protein translocation machinery to import proteins back into the endosymbiont and creation of targeting sequences on the protein substrates themselves. Using a combination of two independent prediction methods, validated through tests on simulated genomes, we show that at least 5% of proteins encoded by an extant proteobacterium are predisposed for targeting to mitochondria, and propose we that mitochondrial targeting information was preexisting for many proteins of the endosymbiont. We analyzed a family of proteins whose members exist both in bacteria and in mitochondria of eukaryotes and show that the amino terminal extensions occasionally found in bacterial family members can function as a crude import sequence when the protein is presented to isolated mitochondria. This activity leaves the development of a primitive translocation channel in the outer membrane of the endosymbiont as a single hurdle to initiating the evolution of mitochondria. PMID- 14739248 TI - Evolutionary genomics of chromoviruses in eukaryotes. AB - The diversity, origin, and evolution of chromoviruses in Eukaryota were examined using the massive amount of genome sequence data for different eukaryotic lineages. A surprisingly large number of novel full-length chromoviral elements were found, greatly exceeding the number of the known chromoviruses. These new elements are mostly structurally intact and highly conserved. Chromoviruses in the key Amniota lineage, the reptiles, have been analyzed by PCR to explain their evolutionary dynamics in amniotes. Phylogenetic analyses provide evidence for a novel centromere-specific chromoviral clade that is widespread and highly conserved in all seed plants. Chromoviral diversity in plants, fungi, and vertebrates, as shown by phylogenetic analyses, was found to be much greater than previously expected. The age of plant chromoviruses has been significantly extended by finding their representatives in the most basal plant lineages, the green and the red algae. The evolutionary origin of chromoviruses has been found to be no earlier than in Cercozoa. The evolutionary history and dynamics of chromoviruses can be explained simply by strict vertical transmission in plants, followed by more complex evolution in fungi and in Metazoa. The currently available data clearly show that chromoviruses indeed represent the oldest and the most widespread clade of Metaviridae. PMID- 14739249 TI - Leptin receptor isoform 219.1: an example of protein evolution by LINE-1-mediated human-specific retrotransposition of a coding SVA element. AB - Phylogenetically new insertions of repetitive sequences may contribute to genome evolution by altering the function of preexisting proteins. One example is the SVA sequence, which forms the C-terminal coding exon of the human leptin receptor isoform 219.1. Here, we report that the SVA insertion into the LEPR locus has occurred after divergence of humans and chimpanzees. The SVA element was inserted into a Hal-1/LINE element present in all monkeys and apes tested. Structural features point toward an integration event that was mediated by the L1 protein machinery acting in trans. Thus, our findings add evidence to the hypothesis that retrotransposition events are a driving force in genomic evolution and that the presence or absence of specific retroelements are one distinguishing feature that separates humans from chimpanzees. PMID- 14739250 TI - Evolution of the multifunctional protein tyrosine phosphatase family. AB - The protein tyrosine phosphatase (PTP) family plays a central role in signal transduction pathways by controlling the phosphorylation state of serine, threonine, and tyrosine residues. PTPs can be divided into dual specificity phosphatases and the classical PTPs, which can comprise of one or two phosphatase domains. We studied amino acid substitutions at functional sites in the phosphatase domain and identified putative noncatalytic phosphatase domains in all subclasses of the PTP family. The presence of inactive phosphatase domains in all subclasses indicates that they were invented multiple times in evolution. Depending on the domain composition, loss of catalytic activity can result in different consequences for the function of the protein. Inactive single-domain phosphatases can still specifically bind substrate and protect it from dephosphorylation by other phosphatases. The inactive domains of tandem phosphatases can be further subdivided. The first class is more conserved, still able to bind phosphorylated tyrosine residues and might recruit multiphosphorylated substrates for the adjacent active domain. The second has accumulated several variable amino acid substitutions in the catalytic center, indicating a complete loss of tyrosine-binding capabilities. To study the impact of substitutions in the catalytic center to the evolution of the whole domain, we examined the evolutionary rates for each individual site and compared them between the classes. This analysis revealed a release of evolutionary constraint for multiple sites surrounding the catalytic center only in the second class, emphasizing its difference in function compared with the first class. Furthermore, we found a region of higher conservation common to both domain classes, suggesting a new regulatory center. We discuss the influence of evolutionary forces on the development of the phosphatase domain, which has led to additional functions, such as the specific protection of phosphorylated tyrosine residues, substrate recruitment, and regulation of the catalytic activity of adjacent domains. PMID- 14739251 TI - Differential enzyme targeting as an evolutionary adaptation to herbivory in carnivora. AB - Not all members of the order Carnivora are carnivorous. Some are omnivorous, and a few, such as the giant panda, Ailuropoda melanoleuca, are almost exclusively herbivorous. Although a number of adaptations to increased plant-eating are recognized within Carnivora, few have been studied at the molecular level. One molecular adaptation to diet that is spread widely across Mammalia is the differential intracellular targeting of the intermediary metabolic enzyme alanine:glyoxylate aminotransferase (AGT), which tends to be mitochondrial in carnivores, peroxisomal in herbivores, and both mitochondrial and peroxisomal in omnivores. In the present study, we have analyzed the targeting of AGT in Carnivora in relation to species' natural diets. We show not only that there has been an adaptive shift in AGT targeting from the mitochondrion toward the peroxisome as diets have shifted from being mainly carnivorous to ones that are more omnivorous and herbivorous but also that in one lineage, namely that of the giant panda, there is evidence for positive selection pressure at the molecular level on the AGT mitochondrial targeting sequence to decrease its efficiency, thereby allowing more AGT to be targeted to the peroxisomes. PMID- 14739252 TI - The low evolutionary rate of human T-cell lymphotropic virus type-1 confirmed by analysis of vertical transmission chains. AB - The evolutionary rate of the human T-cell lymphotropic virus type-1 (HTLV-1) is considered to be very low, in strong contrast to the related human retrovirus HIV. However, current estimates of the HTLV-1 rate rely on the anthropological calibration of phylogenies using assumed dates of human migration events. To obtain an independent rate estimate, we analyzed two variable regions of the HTLV 1 genome (LTR and env) from eight infected families. Remarkable genetic stability was observed, as only two mutations in LTR (756 bp) and three mutations in env (522 bp) occurred within the 16 vertical transmission chains, including one ambiguous position in each region. The evolutionary rate in HTLV-1 was then calculated using a maximum-likelihood approach that used the highest and lowest possible times of HTLV-1 shared ancestry, given the known transmission histories. The rates for the LTR and env regions were 9.58 x 10(-8)-1.25 x 10(-5) and 7.84 x 10(-7) -2.33 x 10(-5)nucleotide substitutions per site per year, respectively. A more precise estimate was obtained for the combined LTR-env data set, which was 7.06 x 10(-7)-1.38 x 10(-5)substitutions per site per year. We also note an interesting correlation between the occurrence of mutations in HTLV-1 and the age of the individual infected. PMID- 14739253 TI - Are combined analyses better than single gene phylogenies? A case study using SSU rDNA and rbcL sequence comparisons in the Zygnematophyceae (Streptophyta). AB - Although the combination of different genes in phylogenetic analyses is a promising approach, the methodology is not well established and analyses often suffer from inadequate, noncongruent taxon sampling, long-branch attraction, or conflicting evolutionary models of the genes analyzed. Conflicts or congruence between multigene and single-gene phylogenies, as well as the assumed superiority of the multigene approach, are often difficult to assess solely because of incongruent taxon sampling. In the present study, a data set of 43 nuclear encoded SSU rDNA and plastid-encoded rbcL gene sequences was generated from the same strains of conjugating green algae (Zygnematophyceae, Streptophyta). Phylogenetic analyses used the genes individually and in combination, either as concatenated sequences or with the log-likelihood summation method. Single-gene analyses, although mostly congruent, revealed some conflicting nodes and showed different patterns of statistical support. Combined analyses confidently resolved the conflicts between the single-gene analyses, enhanced phylogenetic resolution, and were better supported by morphological information. Long-branch taxa were not the same for the two genes analyzed, and, thus, their effect on phylogenetic resolution was minimized in the combined analyses. PMID- 14739254 TI - Regulation of the aldo-keto reductase gene akr1b7 by the nuclear oxysterol receptor LXRalpha (liver X receptor-alpha) in the mouse intestine: putative role of LXRs in lipid detoxification processes. AB - Liver X receptors (LXRs) regulate the expression of a number of genes involved in cholesterol and lipid metabolism after activation by their cognate oxysterol ligands. AKR1-B7 (aldo-keto reductase 1-B7) is expressed in LXR target tissues such as intestine, and because of its known role in detoxifying lipid peroxides, we investigated whether the AKR1-B7 detoxification pathway was regulated by LXRs. Here we show that synthetic LXR agonists increase the accumulation of AKR1-B7 mRNA and protein levels in mouse intestine in wild-type but not lxr(-/-) mice. Regulation of akr1b7 by retinoic X receptor/LXR heterodimers is dependent on three response elements in the proximal murine akr1b7 promoter. Two of these cis acting elements are specific for regulation by the LXRalpha isoform. In addition, in duodenum of wild-type mice fed a synthetic LXR agonist, we observed an LXR dependent decrease in lipid peroxidation. Our results demonstrate that akr1b7 is a direct target of LXRs throughout the small intestine, and that LXR activation plays a protective role by decreasing the deleterious effects of lipid peroxides in duodenum. Taken together, these data suggest a new role for LXRs in lipid detoxification. PMID- 14739255 TI - The glucocorticoid receptor and the orphan nuclear receptor chicken ovalbumin upstream promoter-transcription factor II interact with and mutually affect each other's transcriptional activities: implications for intermediary metabolism. AB - Glucocorticoids exert their metabolic effect via their intracellular receptor, the glucocorticoid receptor (GR). In a yeast two-hybrid screening, we found the chicken ovalbumin upstream promoter transcription factor II (COUP-TFII), an orphan nuclear receptor that plays important roles in glucose, cholesterol, and xenobiotic metabolism, as a partner of GR. In an in vitro glutathione-S transferase pull-down assay, COUP-TFII interacted via its DNA-binding domain with the hinge regions of both GRalpha and its splicing variant GRbeta, whereas COUP TFII formed a complex with GRalpha, but not with GRbeta, in an in vivo chromatin immunoprecipitation and a regular immunoprecipitation assay. Accordingly, GRalpha, but not GRbeta, enhanced COUP-TFII-induced transactivation of the simple COUP-TFII-responsive 7alpha-hydroxylase promoter through the transcriptional activity of its activation function-1 domain, whereas COUP-TFII repressed GRalpha induced transactivation of the glucocorticoid-responsive promoter by attracting the silencing mediator for retinoid and thyroid hormone receptors. Importantly, mutual protein-protein interaction of GRalpha and COUP-TFII was necessary for glucocorticoid-induced enhancement of the promoter activity and the endogenous mRNA expression of the COUP-TFII-responsive phosphoenolpyruvate carboxykinase, the rate-limiting enzyme of hepatic gluconeogenesis. We suggest that COUP-TFII may participate in some of the metabolic effects of glucocorticoids through direct interactions with GRalpha. These interactions influence the transcription of both COUP-TFII- and GRalpha-responsive target genes, seem to be promoter specific, and can be in either a positive or negative direction. PMID- 14739256 TI - Overexpression of follistatin-like 3 in gonads causes defects in gonadal development and function in transgenic mice. AB - Activin has numerous biological activities including regulation of follicular development, spermatogenesis, and steroidogenesis within the gonads. Activities of activin are regulated by follistatin (FST), an activin binding protein, and perhaps follistatin-like 3 (FSTL3; also known as FLRG and FSRP). FSTL3 is a recently described member of the FST family having an overall structure and activity profile similar to that of FST, including binding and neutralization of activin. FSTL3 is most highly expressed in the placenta and testis, whereas FST is highest in the ovary and kidney, suggesting that FSTL3 has biological actions that do not entirely overlap those of FST. To investigate the role of local FSTL3 as a potential regulator of activin action in gonad development and function, we examined FSTL3 expression in the mouse testis. FSTL3 protein was localized to Leydig cells, spermatagonia, and mature spermatids in normal male mice. We then created transgenic mice using a human FSTL3 cDNA driven by the mouse alpha inhibin promoter. Three of five transgenic founders were fertile and were bred to establish lines. In the highest expressing line 3, transgene expression was largely restricted to gonads, with pituitary, adrenal, brain, and uterine expression being substantially lower. Gonad weights, sperm counts, and fertility were significantly reduced in transgenic males, and reduced litter size was evident in line 3 females. Within the testis, highest transgene expression was observed in Sertoli cells, and although most tubules showed evidence of normal spermatogenic development, degenerating tubules devoid of germ cells and Leydig cell hyperplasia were also evident in every line 3 animal examined. Ovaries from line 3 females contained fewer antral follicles and more apparent follicular atresia. Although circulating human FSTL3 levels were undetectable, FSH and LH levels in adult transgenic mice were not significantly different from wild-type animals. However, testosterone levels were significantly increased at d 21 and significantly reduced at d 60 compared with wild-type males. These results suggest that FSTL3 is likely to be a local regulator of activin action in gonadal development and gametogenesis and, further, that activin appears to have important actions in gonadal development and function that are critical for normal reproduction. PMID- 14739257 TI - A high-throughput approach for subcellular proteome: identification of rat liver proteins using subcellular fractionation coupled with two-dimensional liquid chromatography tandem mass spectrometry and bioinformatic analysis. AB - Four fractions from rat liver (a crude mitochondria (CM) and cytosol (C) fraction obtained with differential centrifugation, a purified mitochondrial (PM) fraction obtained with nycodenz density gradient centrifugation, and a total liver (TL) fraction) were analyzed with two-dimensional liquid chromatography tandem mass spectrometry analysis. A total of 564 rat proteins were identified and were bioinformatically annotated according to their physicochemical characteristics and functions. While most extreme alkaline ribosomal proteins were identified in the TL fraction, the C fraction mainly included neutral enzymes and the PM fraction enriched alkaline proteins and proteins with electron transfer activity or oxygen binding activity. Such characteristics were more apparent in proteins identified only in the TL, C, or PM fraction. The Swiss-Prot annotation and the bioinformatic prediction results proved that the C and PM fractions had enriched cytoplasmic or mitochondrial proteins, respectively. Combination usage of subcellular fractionation with two-dimensional liquid chromatography tandem mass spectrometry was proved to be a high-throughput, sensitive, and effective analytical approach for subcellular proteomics research. Using such a strategy, we have constructed the largest proteome database to date for rat liver (564 rat proteins) and its cytosol (222 rat proteins) and mitochondrial fractions (227 rat proteins). Moreover, the 352 proteins with Swiss-Prot subcellular location annotation in the 564 identified proteins were used as an actual subcellular proteome dataset to evaluate the widely used bioinformatics tools such as PSORT, TargetP, TMHMM, and GRAVY. PMID- 14739258 TI - An approach to the genetics of nitrogen use efficiency in maize. AB - To study the genetic variability and the genetic basis of nitrogen (N) use efficiency in maize, a set of recombinant inbred lines crossed with a tester was studied at low input (N-) and high input (N+) for grain yield and its components, grain protein content, and post-anthesis nitrogen uptake and remobilization. Other physiological traits, such as nitrate content, nitrate reductase, glutamine synthetase (GS), and glutamate dehydrogenase activities were studied at the level of the lines. Genotypexnitrogen (GxN) interaction was significant for yield and explained by variation in kernel number. In N-, N-uptake, the nitrogen nutrition index, and GS activity in the vegetative stage were positively correlated with grain yield, whereas leaf senescence was negatively correlated. Whatever N-input, post-anthesis N-uptake was highly negatively related to N-remobilization. As a whole, genetic variability was expressed differently in N+ and N-. This was confirmed by the detection of QTLs. More QTLs were detected in N+ than in N- for traits of vegetative development, N-uptake, and grain yield and its components, whereas it was the reverse for grain protein content and N-utilization efficiency. Several coincidences between genes encoding for enzymes of N metabolism and QTLs for the traits studied were observed. In particular, coincidences in three chromosome regions of QTLs for yield and N-remobilization, QTLs for GS activity and a gene encoding cytosolic GS were observed. This may have a physiological meaning. The GS locus on chromosome 5 appears to be a good candidate gene which can, at least partially, explain the variation in nitrogen use efficiency. PMID- 14739259 TI - Phylogenetic variation in the shoot mineral concentration of angiosperms. AB - The calcium (Ca) concentration of plant shoot tissues varies systematically between angiosperm orders. The phylogenetic variation in the shoot concentration of other mineral nutrients has not yet been described at an ordinal level. The aims of this study were (1) to quantify the shoot mineral concentration of different angiosperm orders, (2) to partition the phylogenetic variation in shoot mineral concentration between and within orders, (3) to determine if the shoot concentration of different minerals are correlated across angiosperm species, and (4) to compare experimental data with published ecological survey data on 81 species sampled from their natural habitats. Species, selected pro rata from different angiosperm orders, were grown in a hydroponic system under a constant external nutrient regime. Shoots of 117 species were sampled during vegetative growth. Significant variation in shoot carbon (C), calcium (Ca), potassium (K), and magnesium (Mg) concentration occurred between angiosperm orders. There was no evidence for systematic differences in shoot phosphorus (P) or organic-nitrogen (N) concentration between orders. At a species level, there were strong positive correlations between shoot Ca and Mg concentration, between shoot P and organic-N concentration, and between shoot K concentration and shoot fresh weight:dry weight ratio. Shoot C and cation concentration correlated negatively at a species level. Species within the Poales and the Caryophyllales had distinct shoot mineralogies in both the designed experiment and in the ecological survey. PMID- 14739260 TI - Regulation of K+ channel activities in plants: from physiological to molecular aspects. AB - Plant voltage-gated channels belonging to the Shaker family participate in sustained K+ transport processes at the cell and whole plant levels, such as K+ uptake from the soil solution, long-distance K+ transport in the xylem and phloem, and K+ fluxes in guard cells during stomatal movements. The attention here is focused on the regulation of these transport systems by protein-protein interactions. Clues to the identity of the regulatory mechanisms have been provided by electrophysiological approaches in planta or in heterologous systems, and through analogies with their animal counterparts. It has been shown that, like their animal homologues, plant voltage-gated channels can assemble as homo- or heterotetramers associating polypeptides encoded by different Shaker genes, and that they can bind auxiliary subunits homologous to those identified in mammals. Furthermore, several regulatory processes (involving, for example, protein kinases and phosphatases, G proteins, 14-3-3s, or syntaxins) might be common to plant and animal Shakers. However, the molecular identification of plant channel partners is still at its beginning. This paper reviews current knowledge on plant K+ channel regulation at the physiological and molecular levels, in the light of the corresponding knowledge in animal cells, and discusses perspectives for the deciphering of regulatory networks in the future. PMID- 14739261 TI - Breeding for micronutrients in staple food crops from a human nutrition perspective. AB - Over three billion people are currently micronutrient (i.e. micronutrient elements and vitamins) malnourished, resulting in egregious societal costs including learning disabilities among children, increased morbidity and mortality rates, lower worker productivity, and high healthcare costs, all factors diminishing human potential, felicity, and national economic development. Nutritional deficiencies (e.g. iron, zinc, vitamin A) account for almost two thirds of the childhood death worldwide. Most of those afflicted are dependent on staple crops for their sustenance. Importantly, these crops can be enriched (i.e. 'biofortified') with micronutrients using plant breeding and/or transgenic strategies, because micronutrient enrichment traits exist within their genomes that can to used for substantially increasing micronutrient levels in these foods without negatively impacting crop productivity. Furthermore, 'proof of concept' studies have been published using transgenic approaches to biofortify staple crops (e.g. high beta-carotene 'golden rice' grain, high ferritin-Fe rice grain, etc). In addition, micronutrient element enrichment of seeds can increase crop yields when sowed to micronutrient-poor soils, assuring their adoption by farmers. Bioavailability issues must be addressed when employing plant breeding and/or transgenic approaches to reduce micronutrient malnutrition. Enhancing substances (e.g. ascorbic acid, S-containing amino acids, etc) that promote micronutrient bioavailability or decreasing antinutrient substances (e.g. phytate, polyphenolics, etc) that inhibit micronutrient bioavailability, are both options that could be pursued, but the latter approach should be used with caution. The world's agricultural community should adopt plant breeding and other genetic technologies to improve human health, and the world's nutrition and health communities should support these efforts. Sustainable solutions to this enormous global problem of 'hidden hunger' will not come without employing agricultural approaches. PMID- 14739262 TI - Overexpression of NtHAL3 genes confers increased levels of proline biosynthesis and the enhancement of salt tolerance in cultured tobacco cells. AB - The Hal3 protein of Saccharomyces cerevisiae inhibits the activity of PPZ1 type-1 protein phosphatases and functions as a regulator of salt tolerance and cell cycle control. In plants, two HAL3 homologue genes in Arabidopsis thaliana, AtHAL3a and AtHAl3b, have been isolated and the function of AtHAL3a has been investigated through the use of transgenic plants. Expressions of both AtHAL3 genes are induced by salt stress. AtHAL3a overexpressing transgenic plants exhibit improved salt and sorbitol tolerance. In vitro studies have demonstrated that AtHAL3 protein possessed 4'-phosphopantothenoylcysteine decarboxylase activity. This result suggests that the molecular function of plant HAL3 genes is different from that of yeast HAL3. To understand the function of plant HAL3 genes in salt tolerance more clearly, three tobacco HAL3 genes, NtHAL3a, NtHAL3b, and NtHAL3c, from Nicotiana tabacum were identified. NtHAL3 genes were constitutively expressed in all organs and under all conditions of stress examined. Overexpression of NtHAL3a improved salt, osmotic, and lithium tolerance in cultured tobacco cells. NtHAL3 genes could complement the temperature-sensitive mutation in the E. coli dfp gene encoding 4'-phosphopantothenoyl-cysteine decarboxylase in the coenzyme A biosynthetic pathway. Cells overexpressing NtHAL3a had an increased intracellular ratio of proline. Taken together, these results suggest that NtHAL3 proteins are involved in the coenzyme A biosynthetic pathway in tobacco cells. PMID- 14739263 TI - Structure and expression profile of the sucrose synthase multigene family in Arabidopsis. AB - The release of the complete genome sequence of Arabidopsis enabled the largest sucrose synthase family described to date, comprising six distinct members, for which expression profiles were not yet available, to be identified. Aimed at understanding the precise function of each AtSUS member among the family, a comparative study of protein structure was performed, together with an expression profiling of the whole gene family using the technique of real-time quantitative reverse transcriptase-polymerase chain reaction. Transcript levels were analysed in several plant organs, including both developing and germinating seeds. A series of treatments such as oxygen deprivation, dehydration, cold treatment, or various sugar feedings were then carried out to characterize the members of the family further. The AtSUS genes exhibit distinct but partially redundant expression profiles. Under anaerobic conditions, for instance, both AtSUS1 and AtSUS4 mRNA levels increase, but in a distinct manner. AtSUS2 is specifically and highly induced in seeds at 12 d after flowering and appears as a marker of seed maturation. AtSUS3 seems to be induced in various organs under dehydration conditions including leaves deprived of water or submitted to osmotic stress as well as late-maturing seeds. AtSUS5 and AtSUS6 are expressed in nearly all plant organs and do not exhibit any transcriptional response to stresses. These results add new insights on the expression of SUS genes and are discussed in relation to distinct functions for each member of the AtSUS family. PMID- 14739264 TI - Gating of water channels (aquaporins) in cortical cells of young corn roots by mechanical stimuli (pressure pulses): effects of ABA and of HgCl2. AB - Hydraulic properties (half-time of water exchange, T1/2, and hydraulic conductivity, Lp; T1/2 approximately 1/Lp) of individual cells in the cortex of young corn roots were measured using a cell pressure probe for up to 6 h to avoid variations between cells. When pulses of turgor pressure of different size were imposed, T1/2 (Lp) responded differently depending on the size. Pulses of smaller than 0.1 MPa, which induced a small proportional water flow, caused no changes in T1/2 (Lp). Medium-sized pulses of between 0.1 and 0.2 MPa caused an increase in T1/2 (decrease in Lp) by a factor of 4 to 23. The effects caused by medium-sized pulses were reversible within 5-20 min. When larger pulses of more than 0.2 MPa were employed, changes were not reversible within 1-3 h, but could be reversed within 30 min in the presence of 500 nM of the stress hormone ABA. Cells with a short T1/2 responded to the aquaporin blocker mercuric chloride (HgCl2). The treatment had no effect on cells which exhibited long T1/2 following a mechanical inhibition by the large-pulse treatment. Step decreases in pressure resulted in the same inhibition as step increases. Hence, the treatment did not cause a stretch-inhibition of water channels and was independent of the directions of both pressure changes and water flows induced by them. It is concluded that inhibition is caused by the absolute value of intensities of water flow within the channels, which increased in proportion to the size of step changes in pressure. Probable mechanisms by which the mechanical stimuli are perceived are (i) the input of kinetic energy to the channel constriction (NPA motif of aquaporin) which may cause a conformational change of the channel protein (energy input model) or (ii) the creation of tensions at the constriction analogous to Bernoulli's principle for macroscopic pores (cohesion-tension model). Estimated rates of water flow within the pores were a few hundred micro m s-1, which is too small to create sufficient tension. They were much smaller than those proposed for AQP1. Based on literature data of single-channel permeability of AQP1, a per channel energy input of 200 kBxT (kB=Boltzmann constant) was estimated for the energy-input model. This should be sufficient to initiate changes of protein conformation and an inactivation of channels. The data indicate different closed states which differ in the amount of distortion and the rates at which they relax back to the open state. PMID- 14739265 TI - Zinnia elegans uses the same peroxidase isoenzyme complement for cell wall lignification in both single-cell tracheary elements and xylem vessels. AB - The nature of the peroxidase isoenzyme complement responsible for cell wall lignification in both Zinnia elegans seedlings and Z. elegans tracheary single cell cultures have been studied. Results showed that both hypocotyls and stems from lignifying Z. elegans seedlings express a cell wall-located basic peroxidase of pI approximately 10.2, which was purified to homogeneity. Molecular mass determination under non-denaturing conditions showed an M(r) of about 43 000, similar to that of other plant peroxidases. The purified Z. elegans peroxidase showed absorption maxima at 403 (Soret band), and at 496-501 and 632-635 (alpha and beta absorption bands), indicating that this enzyme is a high spin ferric haem protein, belonging to the plant peroxidase superfamily, the prosthetic group being ferric protoporphyrin IX. The N-terminal amino acid sequence of this Z. elegans basic peroxidase was KVAVSPLS (peptide motif in bold), which shows strong homologies with the N-amino acid terminus of other strongly basic plant peroxidases. Isoenzyme and western blot analyses showed that this peroxidase isoenzyme is also expressed in trans-differentiating Z. elegans tracheary single cell cultures. The results also showed that Z. elegans tracheary single-cell cultures not only express the same peroxidase isoenzyme as the Z. elegans lignifying xylem, but that this peroxidase isoenzyme acts as a marker of tracheary element differentiation in Z. elegans mesophyll single-cell cultures. From these results, it may be concluded that Z. elegans uses a single programme, i.e. an identical peroxidase isoenzyme complement, for lignification of the xylem, regardless of the existence of different ontogenesis pathways from either mesophyll cells (in the case of tracheary elements) or cambial derivatives (in the case of xylem vessels). PMID- 14739266 TI - Water permeability and reflection coefficient of the outer part of young rice roots are differently affected by closure of water channels (aquaporins) or blockage of apoplastic pores. AB - The relative contribution of the apoplastic and cell-to-cell paths to the overall hydraulic conductivity of the outer part of rice roots (LpOPR) was estimated using a pressure perfusion technique for 30-d-old rice plants (lowland cultivar, IR64, and upland cultivar, Azucena). The technique was based on the perfusion of aerenchyma of root segments from two different zones (20-50 mm and 50-100 mm from the root apex) with aerated nutrient solution using precise pump rates. The outer part of roots (OPR) comprised an outermost rhizodermis, an exodermis, sclerenchyma fibre cells, and the innermost unmodified cortical cell layer. No root anatomical differences were observed for the two cultivars used. Development of apoplastic barriers such as Casparian bands and suberin lamellae in the exodermis were highly variable. On average, matured apoplastic barriers were observed at around 50-70 mm from the root apex. Lignification of the exodermis was completed earlier than that of sclerenchyma cells. Radial water flow across the OPR was impeded either by partially blocking off the porous apoplast with China ink particles (diameter 50 nm) or by closing water channels (aquaporins) in cell membranes with 50 micro M HgCl2. The reduction of LpOPR was relatively larger in the presence of an apoplastic blockage with ink ( approximately 30%) than in the presence of the water channel blocker ( approximately 10%) suggesting a relatively larger apoplastic water flow. The reflection coefficient of the OPR (sigmasOPR) for mannitol significantly increased during both treatments. It was larger when pores of the apoplast were closed, but absolute values were low (overall range of sigmasOPR=0.1-0.4), which also suggested a large contribution of the non-selective, apoplastic path to overall water flow. The strongest evidence in favour of a predominantly apoplastic water transport came from the comparison between diffusional (PdOPR, measured with heavy water, HDO) and osmotic water permeability (PfOPR) or hydraulic conductivity (LpOPR). PfOPR was larger by a factor of 600-1400 compared with P(dOPR). The development of OPR along roots resulted in a decrease of PdOPR by a factor of three (segments taken at 20-50 and 50-100 mm from root apex, respectively). Heat-killing of living cells resulted in an increase of PdOPR for both immature (20-50 mm) and mature (50-100 mm) root segments by a factor of two. Even though both pathways (apoplast and cell-to-cell) contributed to the overall water flow, the findings indicate predominantly apoplastic water flow across the OPR, even in the presence of apoplastic barriers. Low diffusional water permeabilities may suggest a low rate of oxygen diffusion across the OPR from aerenchyma to the outer anaerobic soil medium (low PO2OPR). To date, there are no data on PO2OPR. Provisional data of radial oxygen losses (ROL) across the OPR suggest that, unlike water, rice roots efficiently retain oxygen within the aerenchyma. This ability strongly increases as roots/OPR develop. PMID- 14739267 TI - A cohesion/tension mechanism explains the gating of water channels (aquaporins) in Chara internodes by high concentration. AB - Isolated internodes of Chara corallina have been used to study the gating of aquaporins (water channels) in the presence of high concentrations of osmotic solutes of different size (molecular weight). Osmolytes were acetone and three glycol ethers: ethylene glycol monomethyl ether (EGMME), diethylene glycol monomethyl ether (DEGMME), and triethylene glycol monoethyl ether (TEGMEE). The 'osmotic efficiency' of osmolytes was quite different. Their reflection coefficients ranged between 0.15 (acetone), 0.59 (EGMME), 0.78 (DEGMME), and 0.80 (TEGMEE). Bulk water permeability (Lp) and diffusive permeabilities (Ps) of heavy water (HDO), hydrogen peroxide (H2O2), acetone, and glycol ethers (EGMME, DEGMME, and TEGMEE) were measured using a cell pressure probe. Cells were treated with different concentrations of osmotic solutes of up to 800 mM ( approximately 2.0 MPa of osmotic pressure). Inhibition of aquaporin activity increased with both increasing concentration and size of solutes (reflection coefficients). As cell Lp decreased, Ps increased, indicating that water and solutes used different passages across the plasma membrane. Similar to earlier findings of an osmotic gating of ion channels, a cohesion/tension model of the gating of water channels in Chara internodes by high concentration is proposed. According to the model, tensions (negative pressures) within water channels affected the open/closed state by changing the free energy between states and favoured a distorted/collapsed rather than the open state. They should have differed depending on the concentration and size of solutes that are more or less excluded from aquaporins. The bigger the solute, the lower was the concentration required to induce a reversible closure of aquaporins, as predicted by the model. PMID- 14739268 TI - Actin-dependent fluid-phase endocytosis in inner cortex cells of maize root apices. AB - The fluorescent dye Lucifer Yellow (LY) is a well-known and widely-used marker for fluid-phase endocytosis. In this paper, both light and electron microscopy revealed that LY was internalized into transition zone cells of the inner cortex of intact maize root apices. The internalized LY was localized within tubulo vesicular compartments invaginating from the plasma membrane at actomyosin enriched pit-fields and individual plasmodesmata, as well as within adjacent small peripheral vacuoles. The internalization of LY was blocked by pretreating the roots with the F-actin depolymerizing drug latrunculin B, but not with the F actin stabilizer jasplakinolide. F-actin enriched plasmodesmata and pit-fields of the inner cortex also contain abundant plant-specific unconventional class VIII myosin(s). In addition, 2,3 butanedione monoxime, a general inhibitor of myosin ATPases, partially inhibited the uptake of LY into cells of the inner cortex. Conversely, loss of microtubules did not inhibit fluid-phase endocytosis of LY into these cells. In conclusion, specialized actin- and myosin VIII-enriched membrane domains perform a tissue-specific form of fluid-phase endocytosis in maize root apices. The possible physiological relevance of this process is discussed. PMID- 14739269 TI - Catalase activity and expression in developing sunflower seeds as related to drying. AB - Changes in catalase (CAT) activity and in CAT isoform pattern and expression were investigated in developing sunflower (Helianthus annuus L.) seeds during desiccation on the mother plant and after artificial drying on the flowerheads. Seeds regularly desiccated during their development on the mother plant and reached mass maturity at c. 42 d after flowering (DAF). Freshly harvested seeds did not germinate at any stage of development because they were dormant, but their dormancy was broken after 5-6 months of dry storage. Immature seeds were desiccation-tolerant at 24 DAF since they were able to germinate fully after artificial drying on the flowerheads followed by dry storage. CAT activity increased in non-dehydrated seeds during their development, reaching a maximum a little after seed mass maturity and after artificial drying in immature seeds. This stimulation of CAT activity by natural and artificial drying was related to changes in CAT isoform pattern. Of the four constitutive CAT subunits, that of 59 kDa was always present, but dehydration induced the synthesis of a 55 kDa subunit. This synthesis of the CAT 55 kDa subunit resulted from an activation of the CATA1 gene, suggesting that the regulation of catalase activity and synthesis by drying occurred at the transcriptional level. The increase in CAT activity induced by seed drying was associated with a decrease in hydrogen peroxide level and in lipid peroxidation. These results suggest that CAT plays a role during seed desiccation by preventing dehydration-related oxidative damage and that H(2)O(2) may play a role in the regulation of CAT gene expression and the transduction pathway of the dehydration signal. PMID- 14739270 TI - High temperature stress of Brassica napus during flowering reduces micro- and megagametophyte fertility, induces fruit abortion, and disrupts seed production. AB - High temperature stress (HTS), during flowering, decreases seed production in many plants. To determine the effect of a moderate HTS on flowering, fruit and seed set in Brassica napus, plants were exposed to a HTS (8/16 h dark/light, 18 degrees C night, ramped at 2 degrees C h-1, over 6 h, to 35 degrees C for 4 h, ramped at 2 degrees C h-1 back to 23 degrees C for 6 h) for 1 or 2 weeks after the initiation of flowering. Although flowering on the HTS-treated plants, during both the 1 week and 2 week HTS treatments, was equal to that of control-grown plants, fruit and seed development, as well as seed weight, were significantly reduced. Under HTS, flowers either developed into seedless, parthenocarpic fruit or aborted on the stem. At the cessation of the HTS, plants compensated for the lack of fruit and seed production by increasing the number of lateral inflorescences produced. During the HTS, pollen viability and germinability were slightly reduced. In vitro pollen tube growth at 35 degrees C, from both control pollen and pollen developed under a HTS, appeared abnormal, however, in vivo tube growth to the micropyle appeared normal. Reciprocal pollination of HTS or control pistils with HTS or control pollen indicated that the combined effects of HTS on both micro- and megagametophytes was required to knock out fruit and seed development. Expression profiles for a subset of HEAT SHOCK PROTEINs (HSP101, HSP70, HSP17.6) showed that both micro- and megagametophytes were thermosensitive despite HTS-induced expression from these genes. PMID- 14739271 TI - Photoperiod and temperature differentially regulate the expression of two dehydrin genes during overwintering of birch (Betula pubescens Ehrh.). AB - The overwintering of trees in northern areas depends on processes regulated by photoperiod and temperature. To identify the physiological and genetic factors involved in this environmental control, three latitudinal ecotypes of pubescent birch (Betula pubescens Ehrh.) growing in a common garden experiment were used. Each ecotype responded to the shortening of the photoperiod according to its specific critical daylength, resulting in the induction of freezing tolerance and dehydration of buds first in the northern ecotype, followed by the central and southern ecotypes, respectively. By contrast, there was no clear difference in the timing of dormancy release, bud rehydration, and deacclimation in the spring, suggesting that these traits were controlled mainly by temperature. To elucidate the role of dehydrins (DHN) in the overwintering process, two DHN genomic clones were isolated from pubescent birch and expression of the corresponding genes, both in field and under controlled conditions, was characterized. BpuDhn1 was found to encode an Y(n)K(n)-type of basic DHN, while BpuDhn2 encoded an acidic, SK(n)-type of DHN. In field-grown trees the level of BpuDhn1 increased in buds during the autumn, while the level of BpuDhn2 was highest during the coldest winter months. Under controlled conditions BpuDhn1 increased in response to the combined effect of short daylength and low, non-freezing temperatures whereas the expression of BpuDhn2 was mainly controlled by low temperature while photoperiod had less effect on its expression. These results suggest that DHNs participate in the sensitive environmental regulation of the overwintering process in birch. PMID- 14739272 TI - A transgene encoding a blue-light receptor, phot1, restores blue-light responses in the Arabidopsis phot1 phot2 double mutant. AB - Phototropins (phot1 and phot2) are suggested to be multifunctional blue-light (BL) receptors mediating phototropism, chloroplast movement, stomatal opening, and leaf expansion. The Arabidpsis phot1 phot2 double mutant lacks all of these responses. To confirm the requirement of phototropins in BL responses, the Arabidopsis phot1 phot2 double mutant was transformed with PHOT1 cDNA and the phenotypic restoration was analysed in the transformants. It was found that all BL responses were restored, although differentially, by the transformation of the Arabidopsis phot1 phot2 double mutant with PHOT1 cDNA. The results showed that phot1 was an essential component for all these BL responses in planta, and that the cellular level of phot1 might determine the individual BL responses. PMID- 14739273 TI - Impact of temperature on the arbuscular mycorrhizal (AM) symbiosis: growth responses of the host plant and its AM fungal partner. AB - The growth response of the hyphae of mycorrhizal fungi has been determined, both when plant and fungus together and when only the fungus was exposed to a temperature change. Two host plant species, Plantago lanceolata and Holcus lanatus, were grown separately in pots inoculated with the mycorrhizal fungus Glomus mosseae at 20/18 degrees C (day/night); half of the pots were then transferred to 12/10 degrees C. Plant and fungal growth were determined at six sequential destructive harvests. A second experiment investigated the direct effect of temperature on the length of the extra-radical mycelium (ERM) of three mycorrhizal fungal species. Growth boxes were divided in two equal compartments by a 20 micro m mesh, allowing only the ERM and not roots to grow into a fungal compartment, which was either heated (+8 degrees C) or kept at ambient temperature. ERM length (LERM) was determined on five sampling dates. Growth of H. lanatus was little affected by temperature, whereas growth of P. lanceolata increased with temperature, and both specific leaf area (SLA) and specific root length (SRL) increased independently of plant size. Percentage of colonized root (LRC) and LERM were positively correlated with temperature when in symbiosis with P. lanceolata, but differences in LRC were a function of plant biomass. Colonization was very low in H. lanatus roots and there was no significant temperature effect. In the fungal compartment LERM increased over time and was greatest for Glomus mosseae. Heating the fungal compartment significantly increased LERM in two of the three species but did not affect LRC. However, it significantly increased SRL of roots in the plant compartment, suggesting that the fungus plays a regulatory role in the growth dynamics of the symbiosis. These temperature responses have implications for modelling carbon dynamics under global climate change. PMID- 14739274 TI - Proteolytic processing and translation initiation: two independent mechanisms for the expression of the Sendai virus Y proteins. AB - The four Sendai virus C-proteins (C', C, Y1, and Y2) represent an N-terminal nested set of non-structural proteins whose expression modulates both the readout of the viral genome and the host cell response. In particular, they modulate the innate immune response by perturbing the signaling of type 1 interferons. The initiation codons for the four C-proteins have been mapped in vitro, and it has been proposed that the Y proteins are initiated by ribosomal shunting. A number of mutations were reported that significantly enhanced Y expression, and this was attributed to increased shunt-mediated initiation. However, we demonstrate that this arises due to enhanced proteolytic processing of C', an event that requires its very N terminus. Curiously, although Y expression in vitro is mediated almost exclusively by initiation, Y proteins in vivo can arise both by translation initiation and processing of the C' protein. To our knowledge this is the first example of two apparently independent pathways leading to the expression of the same polypeptide chain. This dual pathway explains several features of Y expression. PMID- 14739275 TI - Neuronal calcium sensor protein visinin-like protein-3 interacts with microsomal cytochrome b5 in a Ca2+-dependent manner. AB - Visinin-like protein-3, which is one of the neuronal calcium sensors, has been shown to be mainly expressed in cerebellar Purkinje cells, but cellular function of this protein has not yet been elucidated. We examined the tissue distribution of murine visinin-like protein-3 transcripts using real-time reverse transcription-PCR. Visinin-like protein-3 mRNA was found to be expressed in peripheral tissues. Particularly, the expression of the transcript in the thymus was significantly higher than in other peripheral tissues. In addition, B6RVTC1 thymoma cells robustly expressed visinin-like protein-3 mRNA. To identify a target protein of visinin-like protein-3, we performed a pull-down experiment using glutathione S-transferase-tagged visinin-like protein-3 and two-dimensional electrophoresis. We demonstrated that microsomal cytochrome b(5) was a Ca(2+) dependent binding partner of visinin-like protein-3. In a co-immunoprecipitation experiment, it was observed that hippocalcin, as well as visinin-like protein-3, could interact with cytochrome b(5). Furthermore, we confirmed that the sequence Val(114)-Tyr(127) at the C-terminal tail of cytochrome b(5) is the minimal structural requirement for binding to visinin-like protein-3. In addition, the loop His(19)-His(25) at the N terminus of visinin-like protein-3 is essential for binding to cytochrome b(5). Microsomal cytochrome b(5) was also shown to be a potential activator of cytochrome P450. The present findings raise the possibility that visinin-like protein-3 may link Ca(2+) signaling to the machinery of microsomal monooxygenase complex composed of cytochrome b(5), cytochrome P450, and some reductases. This report provides the first evidence of an interaction between visinin-like protein-3 and microsomal cytochrome b(5). PMID- 14739276 TI - Conformational states of the small G protein Arf-1 in complex with the guanine nucleotide exchange factor ARNO-Sec7. AB - Arf1 is a small G protein involved in vesicular trafficking, and although it is only distantly related to Ras, it adopts a similar three-dimensional structure. In the present work, we study Arf1 bound to GDP and GTP and its interactions with one of its guanosine nucleotide exchange factors, ARNO-Sec7. The (31)P NMR spectra of Arf1.GDP.Mg(2+) and Arf1.GTP.Mg(2+) share the general features typical for all small G proteins studied so far. Especially, the beta-phosphate resonances of the bound nucleotide are shifted strongly downfield compared with the resonance positions of the free magnesium complexes of GDP and GTP. However, no evidence for an equilibrium between two conformational states of Arf1.GDP.Mg(2+) or Arf1.GTP.Mg(2+) could be observed as it was described earlier for Ras and Ran. Glu(156) of ARNO-Sec7 has been suggested to play as "glutamic acid finger" an important role in the nucleotide exchange mechanism. In the millimolar concentration range used in the NMR experiments, wild type ARNO-Sec7 and ARNO-Sec7(E156D) do weakly interact with Arf1.GDP.Mg(2+) but do not form a strong complex with magnesium-free Arf1.GDP. Only wild type ARNO-Sec7 competes weakly with GDP on Arf1.GDP.Mg(2+) and leads to a release of GDP when added to the solution. The catalytically inactive mutants ARNO-Sec7(E156A) and ARNO Sec7(E156K) induce a release of magnesium from Arf1.GDP.Mg(2+) but do not promote GDP release. In addition, ARNO-Sec7 does not interact or only very weakly interacts with the GTP-bound form of Arf1, opposite to the observation made earlier for Ran, where the nucleotide exchange factor RCC1 forms a complex with Ran.GTP.Mg(2+) and is able to displace the bound GTP. PMID- 14739277 TI - Furin is a chemokine-modifying enzyme: in vitro and in vivo processing of CXCL10 generates a C-terminally truncated chemokine retaining full activity. AB - Chemokines comprise a class of structurally related proteins that are involved in many aspects of leukocyte migration under basal and inflammatory conditions. In addition to the large number of genes, limited processing of these proteins by a variety of enzymes enhances the complexity of the total spectrum of chemokine variants. We have recently shown that the native chemokine CXCL10 is processed at the C terminus, thereby shedding the last four amino acids. The present study was performed to elucidate the mechanism in vivo and in vitro and to study the biological activity of this novel isoform of CXCL10. Using a combination of protein purification and mass spectrometric techniques, we show that the production of C-terminally truncated CXCL10 by primary keratinocytes is inhibited in vivo by a specific inhibitor of pro-protein convertases (e.g. furin) but not by inhibition of matrix metalloproteinases. Moreover, CXCL10 is processed by furin in vitro, which is abrogated by a mutation in the furin recognition site. Using GTPgammaS binding, Ca(2+) mobilization, and chemotaxis assays, we demonstrate that the C-terminally truncated CXCL10 variant is a potent ligand for CXCR3. Moreover, the inverse agonist activity on the virally encoded receptor ORF74 and the direct antibacterial activity of CXCL10 are fully retained. Hence, we have identified furin as a novel chemokine-modifying enzyme in vitro and most probably also in vivo, generating a C-terminally truncated CXCL10, which fully retains its (inverse) agonistic properties. PMID- 14739278 TI - Preferential translation of internal ribosome entry site-containing mRNAs during the mitotic cycle in mammalian cells. AB - A cell synchronization protocol was established in which global and individual mRNA translational efficiencies could be examined. While global translational efficiency was reduced in mitotic cells, approximately 3% of mRNAs remained predominantly associated with large polysomes during mitosis, as determined by cDNA microarray analyses. The 5'-non-coding regions of six mRNAs were shown to contain internal ribosome entry sites (IRES). However, not all known mRNAs that contain IRES elements were actively translated during mitosis, arguing that specific IRES sequences are differentially regulated during mitosis. PMID- 14739279 TI - The global transcriptional response to transient cell wall damage in Saccharomyces cerevisiae and its regulation by the cell integrity signaling pathway. AB - In the yeast Saccharomyces cerevisiae, environmental stress conditions that damage the cell wall lead to activation of the so-called "compensatory mechanism," aimed at preserving cell integrity through a remodeling of this extracellular matrix. Here we used DNA microarrays to investigate the molecular basis of this response to two agents that induce transient cell wall damage; namely Congo Red and Zymolyase. Treatment of the cells with these two agents elicited the up-regulation of 132 and 101 genes respectively, the main functional groups among them being involved in cell wall construction and metabolism. The main response does not occur until hours after exposure to the cell wall perturbing agent. In some cases, this response was transient, but more sustained in others, especially in the case of the genes involved in cell wall remodeling. Clustering of these data together with those from the response to constitutive cell wall damage, revealed the existence of a cluster of co-regulated genes that was strongly induced under all conditions assayed. Those genes induced by cell wall damage showed an enrichment in DNA binding motifs for Rlm1p, Crz1p, SBF (Swi4p/Swi6p), Msn2p/Msn4p, Ste12p, and Tec1p transcription factors, suggesting a complex regulation of this response together with the possible involvement of several signaling pathways. With the exception of PHO89 and FKS2, none of the genes induced by Congo Red was up-regulated in a slt2 strain. Moreover, characterization of the transcriptional response to Congo Red in a rlm1 mutant strain revealed that only a few genes (i.e. PHO89, FKS2, YLR042C, and CHA1) were induced at least partially independently of the transcription factor Rlm1p, the rest being totally dependent on this transcription factor for their activation. Our findings consistently demonstrate that the cell integrity signaling pathway regulates the cell wall damage compensatory response, mainly through transcriptional activation mediated by Rlm1p. PMID- 14739280 TI - Identification and characterization of a novel human myeloid inhibitory C-type lectin-like receptor (MICL) that is predominantly expressed on granulocytes and monocytes. AB - Inhibitory and activatory C-type lectin-like receptors play an important role in immunity through the regulation of leukocytes. Here, we report the identification and characterization of a novel myeloid inhibitory C-type lectin-like receptor (MICL) whose expression is primarily restricted to granulocytes and monocytes. This receptor, which contains a single C-type lectin-like domain and a cytoplasmic immunoreceptor tyrosine-based inhibitory motif, is related to LOX-1 (lectin-like receptor for oxidized low density lipoprotein-1) and the beta-glucan receptor (Dectin-1) and is variably spliced and highly N-glycosylated. We demonstrate that it preferentially associates with the signaling phosphatases SHP 1 and SHP-2, but not with SHIP. Novel chimeric analyses with a construct combining MICL and the beta-glucan receptor show that MICL can inhibit cellular activation through its cytoplasmic immunoreceptor tyrosine-based inhibitory motif. These data suggest that MICL is a negative regulator of granulocyte and monocyte function. PMID- 14739281 TI - Helix orientation of the functional domains in apolipoprotein e in discoidal high density lipoprotein particles. AB - Human apolipoprotein E (apoE) mediates high affinity binding to the low density lipoprotein receptor when present on a lipidated complex. In the absence of lipid, however, apoE does not bind the receptor. Whereas the x-ray structure of lipid-free apoE3 N-terminal (NT) domain is known, the structural organization of its lipid-associated, receptor-active conformation is poorly understood. To study the organization of apoE amphipathic alpha-helices in a lipid-associated state, single tryptophan-containing apoE3 variants were employed in fluorescence quenching studies. The relative positions of the Trp residues with respect to the phospholipid component of apoE/lipid particles were established from the degree of quenching by phospholipids bearing nitroxide groups at various positions along their fatty acyl chains. Four apoE3-NT variants bearing Trp reporter groups at positions 141, 148, 155, or 162 within helix 4 and two apoE3 variants containing single Trp at positions 257 or 264 in the C-terminal (CT) domain, were reconstituted into phospholipid-containing discoidal complexes. Parallax analysis revealed that each engineered Trp residue in helix 4 of apoE3-NT, as well as those in the CT domain of apoE, localized approximately 5 A from the center of the bilayer. Circular dichroism studies revealed that lipid association induces additional helix formation in apoE. Protease protection assays suggest the flexible loop segment between the NT and CT domains may transition from unstructured to helix upon lipid association. Taken together, these data support a model wherein the alpha-helices in the receptor-binding region and the CT domain of apoE align perpendicular to the fatty acyl chains of the phospholipid bilayer. In this alignment, the residues of helix 4 are arrayed in a positively charged, curved helical segment for optimal receptor interaction. PMID- 14739282 TI - The role of the C-terminal extension (CTE) of the estrogen receptor alpha and beta DNA binding domain in DNA binding and interaction with HMGB. AB - HMGB-1/-2 are coregulatory proteins that facilitate the DNA binding and transcriptional activity of steroid receptor members of the nuclear receptor family of transcription factors. We investigated the influence and mechanism of action of HMGB-1/-2 (formerly known as HMG-1/-2) on estrogen receptor alpha (ERalpha) and ERbeta. Both ER subtypes were responsive to HMGB-1/-2 with respect to enhancement of receptor DNA binding affinity and transcriptional activity in cells. Responsiveness to HMGB-1/-2 was dependent on the C-terminal extension (CTE) region of the ER DNA binding domain (DBD) and correlated with a direct protein interaction between HMGB-1/-2 and the CTE. Thus the previously reported higher DNA binding affinity and transcription activity of ERalpha as compared with ERbeta is not due to a lack of ERbeta interaction with HMGB-1/-2. Using chimeric receptor DBDs, the higher intrinsic DNA binding affinity of ERalpha than ERbeta was shown to be due to a unique property of the ERalpha CTE, independent of HMGB-1/-2. The CTE of both ER subtypes was also shown to be required for interaction with ERE half-sites. These studies reveal the importance of the CTE and HMGB-1/-2 for ERalpha and ERbeta interaction with their cognate target DNAs. PMID- 14739283 TI - Voltage-dependent anion-selective channels VDAC2 and VDAC3 are abundant proteins in bovine outer dense fibers, a cytoskeletal component of the sperm flagellum. AB - Outer dense fibers (ODF) are specific subcellular components of the sperm flagellum. The functions of ODF have not yet been clearly elucidated. We have investigated the protein composition of purified ODF from bovine spermatozoa and found that one of the most abundant proteins is a 30-32-kDa polypeptide. This protein was analyzed by sequencing peptides derived following limited proteolysis. Peptide sequences were found to match VDAC2 and VDAC3. VDACs (voltage-dependent, anion-selective channels) or eukaryotic porins are a group of proteins first identified in the mitochondrial outer membrane that are able to form hydrophilic pore structures in membranes. In mammals, three VDAC isoforms (VDAC1, -2, -3) have been identified by cDNA cloning and sequencing. Antibodies against synthetic peptides specific for the three mammal VDAC isoforms were generated in rabbits. Their specificity was demonstrated by immunoblotting using recombinant VDAC1, -2, and -3. In protein extracts of bovine spermatozoa, VDAC1, 2, and -3 were detected by specific antibodies, while only VDAC2 and -3 were found as solubilized proteins derived from purified bovine ODFs. Immunofluorescence microscopy of spermatozoa revealed that anti-VDAC2 and anti VDAC3 antibodies clearly bound to the sperm flagellum, in particular to the ODF. Transmission electron immunomicroscopy supported the finding that VDAC2 protein is abundant in the ODF. Since the ODF does not have any known membranous structure, it is tempting to speculate that VDAC2 and VDAC3 might have an alternative structural organization and different functions in ODF than in mitochondria. PMID- 14739284 TI - Sorting motifs in the intracellular domain of the low density lipoprotein receptor interact with a novel domain of sorting nexin-17. AB - The low density lipoprotein (LDL) receptor plays a major role in maintaining human plasma cholesterol levels and mutations in the gene cause familial hypercholesterolemia. The LDL receptor (LDLR) pathway has been well characterized, but little is known of proteins involved in its complex intracellular sorting and trafficking. Sorting nexin 17 (SNX17) has recently been implicated in LDLR intracellular trafficking. We show here that endogenous SNX17 is highly expressed in several cell types and is localized partially in early endosomes. We found that the PX domain of SNX17 is required for its endosomal localization but does not interact directly with the LDL receptor. A novel domain containing a FERM-like domain of SNX17 is needed for its interaction with the LDL receptor. Mutations in the NPXY motif of the LDL-receptor cytoplasmic tail that disrupt internalization also disrupt its interaction with SNX17, whereas mutations elsewhere had little effect. When transiently overexpressed in Chinese hamster ovary cells, SNX17 localized to large vesicular structures and disrupted normal trafficking of the LDL receptor in a PX domain-dependent manner. These results suggest that SNX17 plays a role in the cellular trafficking of the LDL receptor through interaction with the NPVY motif in its cytoplasmic domain and interaction of the PX domain with subcellular membrane compartments. PMID- 14739285 TI - Fatty acids regulate pigmentation via proteasomal degradation of tyrosinase: a new aspect of ubiquitin-proteasome function. AB - Fatty acids are common components of biological membranes that are known to play important roles in intracellular signaling. We report here a novel mechanism by which fatty acids regulate the degradation of tyrosinase, a critical enzyme associated with melanin biosynthesis in melanocytes and melanoma cells. Linoleic acid (unsaturated fatty acid, C18:2) accelerated the spontaneous degradation of tyrosinase, whereas palmitic acid (saturated fatty acid, C16:0) retarded the proteolysis. The linoleic acid-induced acceleration of tyrosinase degradation could be abrogated by inhibitors of proteasomes, the multicatalytic proteinase complexes that selectively degrade intracellular ubiquitinated proteins. Linoleic acid increased the ubiquitination of many cellular proteins, whereas palmitic acid decreased such ubiquitination, as compared with untreated controls, when a proteasome inhibitor was used to stabilize ubiquitinated proteins. Immunoprecipitation analysis also revealed that treatment with fatty acids modulated the ubiquitination of tyrosinase, i.e. linoleic acid increased the amount of ubiquitinated tyrosinase whereas, in contrast, palmitic acid decreased it. Furthermore, confocal immunomicroscopy showed that the colocalization of ubiquitin and tyrosinase was facilitated by linoleic acid and diminished by palmitic acid. Taken together, these data support the view that fatty acids regulate the ubiquitination of tyrosinase and are responsible for modulating the proteasomal degradation of tyrosinase. In broader terms, the function of the ubiquitin-proteasome pathway might be regulated physiologically, at least in part, by fatty acids within cellular membranes. PMID- 14739286 TI - SRP-2 is a cross-class inhibitor that participates in postembryonic development of the nematode Caenorhabditis elegans: initial characterization of the clade L serpins. AB - High molecular weight serpins are members of a large superfamily of structurally conserved proteins that inactivate target proteinases by a suicide substrate-like mechanism. In vertebrates, different clades of serpins distribute predominantly to either the intracellular or extracellular space. Although much is known about the function, structure, and inhibitory mechanism of circulating serpins such as alpha(1)-antitrypsin (SERPINA1) and antithrombin III (SERPINC1), relatively little is known about the function of the vertebrate intracellular (clade B) serpins. To gain a better understanding of the biology of the intracellular serpins, we initiated a comparative genomics study using Caenorhabditis elegans as a model system. A screen of the C. elegans genomic and cDNA databases revealed nine serpin genes, tandemly arrayed on chromosome V. Although the C. elegans serpins represent a unique clade (L), they share significant functional homology with members of the clade B group of intracellular serpins, since they lack typical N-terminal signal peptides and reside intracellularly. To determine whether nematode serpins function as proteinase inhibitors, one family member, srp-2, was chosen for further characterization. Biochemical analysis of recombinant SRP-2 protein revealed SRP-2 to be a dual cross-class inhibitor of the apoptosis-related serine proteinase, granzyme B, and the lysosomal cysteine proteinases, cathepsins K, L, S, and V. Analysis of temporal and spatial expression indicated that SRP-2 was present during early embryonic development and highly expressed in the intestine and hypoderm of larval and adult worms. Transgenic animals engineered to overexpress SRP-2 were slow growing and/or arrested at the first, second, or third larval stages. These data suggest that perturbations of serpin-proteinase balance are critical for correct postembryonic development in C. elegans. PMID- 14739287 TI - Regulation of phospholipid synthesis in the yeast cki1Delta eki1Delta mutant defective in the Kennedy pathway. The Cho1-encoded phosphatidylserine synthase is regulated by mRNA stability. AB - In the yeast Saccharomyces cerevisiae, the most abundant phospholipid phosphatidylcholine is synthesized by the complementary CDP-diacylglycerol and Kennedy pathways. Using a cki1Delta eki1Delta mutant defective in choline kinase and ethanolamine kinase, we examined the consequences of a block in the Kennedy pathway on the regulation of phosphatidylcholine synthesis by the CDP diacylglycerol pathway. The cki1Delta eki1Delta mutant exhibited increases in the synthesis of phosphatidylserine, phosphatidylethanolamine, and phosphatidylcholine via the CDP-diacylglycerol pathway. The increase in phospholipid synthesis correlated with increased activity levels of the CDP diacylglycerol pathway enzymes phosphatidylserine synthase, phosphatidylserine decarboxylase, phosphatidylethanolamine methyltransferase, and phospholipid methyltransferase. However, other enzyme activities, including phosphatidylinositol synthase and phosphatidate phosphatase, were not affected in the cki1Delta eki1Delta mutant. For phosphatidylserine synthase, the enzyme catalyzing the committed step in the pathway, activity was regulated by increases in the levels of mRNA and protein. Decay analysis of CHO1 mRNA indicated that a dramatic increase in transcript stability was a major component responsible for the elevated level of phosphatidylserine synthase. These results revealed a novel mechanism that controls phospholipid synthesis in yeast. PMID- 14739288 TI - Identification and characterization of human glucose transporter-like protein-9 (GLUT9): alternative splicing alters trafficking. AB - The recently cloned human GLUT9 gene, which maps to chromosome 4p15.3-p16, consists of 12 exons coding for a 540-amino acid protein. Based on a sequence entry (NCBI accession number BC018897) and screening of expressed sequence tags, we have cloned an alternative splice variant of GLUT9 from human kidney cDNA. The RNA of this splice variant consists of 13 exons and codes for a putative protein of 512 amino acids (GLUT9DeltaN). The predicted proteins differ only in their N terminus, suggesting a different subcellular localization and possible physiological role. Screening human tissue RNA by reverse transcription-PCR showed that GLUT9 is expressed mainly in kidney, liver, placenta, and leukocytes, whereas GLUT9DeltaN was detected only in kidney and placenta. The GLUT9 protein localized by immunohistochemistry to human kidney proximal tubules, and subcellular fractionation of human kidney revealed the GLUT9 protein in plasma membranes and high density microsomal membranes. Treatment of kidney membrane proteins with peptide N-glycosidase F showed that GLUT9 and GLUT9DeltaN are expressed in vivo. Localization of GLUT9 and GLUT9DeltaN in three kidney-derived cell lines revealed a plasma membrane distribution for GLUT9 in COS-7 and HEK293 cells, whereas GLUT9DeltaN showed a perinuclear pattern and plasma membrane staining in COS-7 and HEK293 cells, respectively. In polarized Madin-Darby canine kidney cells, GLUT9 trafficked to the basolateral membrane, whereas GLUT9DeltaN localized to the apical membrane. Using heterologous expression of GLUT9 in Xenopus oocytes, GLUT9 appears to be a functional isoform with low affinity for deoxyglucose. Deoxyglucose transport mediated by GLUT9 was not inhibited by cytochalasin B. GLUT9 did not bind cytochalasin B as shown by a cytochalasin B binding assay, indicating a similar behavior of GLUT9 compared with GLUT5. PMID- 14739289 TI - tRNA-triggered ATP hydrolysis and generation of membrane potential by the leishmania mitochondrial tRNA import complex. AB - Translocation of tRNAs across mitochondrial membranes is a receptor-mediated active transport process requiring ATP. A large tRNA import complex from the inner membrane of Leishmania mitochondria catalyzes translocation into phospholipid vesicles. In this reconstituted system, the import substrate tRNA(Tyr)(GUA) specifically stimulated hydrolysis of ATP within the vesicles, with the subsequent generation of a membrane potential by pumping out of protons, as shown by the protonophore-sensitive uptake of the potential-sensitive dye rhodamine 123. Generation of membrane potential was dependent on ATP hydrolysis, and inhibited by oligomycin, recalling the proton-translocation mechanism of the respiratory F(1)-F(0)-ATPase. For translocation of tRNA, ATP could be replaced by low pH of the medium, but proton-dependent import was resistant to oligomycin. Moreover, ATP hydrolysis, generation of membrane potential and tRNA uptake were inhibited by carboxyatractyloside, a specific inhibitor of mitochondrial ATP-ADP translocase, implying an ATP requirement within the vesicles. These observations imply a gating mechanism in which tRNA, on binding to its receptor, triggers the energetic activation of the complex, leading to the opening of import channels. PMID- 14739290 TI - Complete inhibition and partial Re-activation of single F1-ATPase molecules by tentoxin: new properties of the re-activated enzyme. AB - During hydrolysis of ATP, the gamma subunit of the rotary motor protein F(1) ATPase rotates within a ring of alpha(3)beta(3) subunits. Tentoxin is a phyto pathogenic cyclic tetrapeptide, which influences F(1)-ATPase activity of sensitive species. At low concentrations, tentoxin inhibits ATP hydrolysis of ensembles of F(1) molecules in solution. At higher concentrations, however, ATP hydrolysis recovers. Here we have examined how tentoxin acts on individual molecules of engineered F(1)-ATPase from the thermophilic Bacillus PS3 (Groth, G., Hisabori, T., Lill, H., and Bald, D. (2002) J. Biol. Chem. 277, 20117-20119). We found that inhibition by tentoxin caused a virtually complete stop of rotation, which was partially relieved at higher tentoxin concentrations. Re activation, however, was not simply a reversal of inhibition; while the torque appears unaffected as compared with the situation without tentoxin, F(1) under re activating conditions was less susceptible to inhibitory ADP binding but displayed a large number of short pauses, indicating infringed energy conversion. PMID- 14739291 TI - Cbfa1/RUNX2 directs specific expression of the sclerosteosis gene (SOST). AB - Loss-of-function mutations in the sclerosteosis gene (SOST) cause a rare sclerosing bone dysplasia characterized by skeletal overgrowth. Cbfa1/RUNX2 is a key transcriptional regulator of osteoblast function. Here we link these two pathways by demonstrating, via gel shift and transient transfection analyses, that Cbfa1 binding to the proximal SOST promoter contributes to differential SOST expression in two osteosarcoma cell lines. Additionally, an E-box binding motif in the 1.8-kb proximal SOST promoter appears to be functional in SAOS-2 cells, but does not account for SAOS-specific expression of SOST. The regulation of SOST expression by Cbfa1 suggests a potential role for the sclerosteosis gene in homeostatic regulation of osteoblast differentiation and function. Furthermore, the juxtaposition of Cbfa1, E-box, and C/EBP binding sites in the SOST proximal promoter bears an intriguing resemblance to the promoter for osteocalcin, another osteoblast-specific gene with a loss-of-function phenotype of bone overgrowth. PMID- 14739292 TI - DNA and RNA binding by the mitochondrial lon protease is regulated by nucleotide and protein substrate. AB - The ATP-dependent Lon protease belongs to a unique group of proteases that bind DNA. Eukaryotic Lon is a homo-oligomeric ring-shaped complex localized to the mitochondrial matrix. In vitro, human Lon binds specifically to a single-stranded GT-rich DNA sequence overlapping the light strand promoter of human mitochondrial DNA (mtDNA). We demonstrate that Lon binds GT-rich DNA sequences found throughout the heavy strand of mtDNA and that it also interacts specifically with GU-rich RNA. ATP inhibits the binding of Lon to DNA or RNA, whereas the presence of protein substrate increases the DNA binding affinity of Lon 3.5-fold. We show that nucleotide inhibition and protein substrate stimulation coordinately regulate DNA binding. In contrast to the wild type enzyme, a Lon mutant lacking both ATPase and protease activity binds nucleic acid; however, protein substrate fails to stimulate binding. These results suggest that conformational changes in the Lon holoenzyme induced by nucleotide and protein substrate modulate the binding affinity for single-stranded mtDNA and RNA in vivo. Co immunoprecipitation experiments show that Lon interacts with mtDNA polymerase gamma and the Twinkle helicase, which are components of mitochondrial nucleoids. Taken together, these results suggest that Lon participates directly in the metabolism of mtDNA. PMID- 14739293 TI - Adhesion or plasmin regulates tyrosine phosphorylation of a novel membrane glycoprotein p80/gp140/CUB domain-containing protein 1 in epithelia. AB - Suspension of cultured human foreskin keratinocytes (HKs) with trypsin phosphorylates tyrosine residues on an 80-kDa membrane glycoprotein, p80 (Xia, Y., Gil, S. G., and Carter, W. G. (1996) J. Cell Biol. 132, 727-740). Readhesion dephosphorylates p80. Sequencing of a p80 cDNA established identity to CUB domain containing protein 1 (CDCP1), a gene elevated in carcinomas. CDCP1/p80 cDNA encodes three extracellular CUB domains, a transmembrane domain, and two putative cytoplasmic Tyr phosphorylation sites. Treatment of adherent HKs with suramin, a heparin analogue, or inhibitors of phosphotyrosine phosphatases (PTPs; vanadate or calpeptin) increases phosphorylation of p80 and a novel 140-kDa membrane glycoprotein, gp140. Phosphorylated gp140 was identified as a trypsin-sensitive precursor to p80. Identity was confirmed by digestion and phosphorylation studies with recombinant gp140-GFP. Plasmin, a serum protease, also converts gp140 to p80, providing biological significance to the cleavage in wounds. Phosphorylation of gp140 and p80 are mediated by Src family kinases at multiple Tyr residues including Tyr(734). Dephosphorylation is mediated by PTP(s). Conversion of gp140 to p80 prolongs phosphorylation of p80 in response to suramin and changes in adhesion. This distinguishes gp140 and p80 and explains the relative abundance of phosphorylated p80 in trypsinized HKs. We conclude that phosphorylation of gp140 is dynamic and balanced by Src family kinase and PTPs yielding low equilibrium phosphorylation. We suggest that the balance is altered by conversion of gp140 to p80 and by adhesion, providing a novel transmembrane phosphorylation signal in epithelial wounds. PMID- 14739294 TI - Systematic peptide engineering and structural characterization to search for the shortest antimicrobial peptide analogue of gaegurin 5. AB - As part of an effort to develop new, low molecular mass peptide antibiotics, we searched for the shortest bioactive analogue of gaegurin 5 (GGN5), a 24-residue antimicrobial peptide. Thirty-one kinds of GGN5 analogues were synthesized, and their biological activities were analyzed against diverse microorganisms and human erythrocytes. The structural properties of the peptides in various solutions were characterized by spectroscopic methods. The N-terminal 13 residues of GGN5 were identified as the minimal requirement for biological activity. The helical stability, the amphipathic property, and the hydrophobic N terminus were characterized as the important structural factors driving the activity. To develop shorter antibiotic peptides, amino acid substitutions in an inactive 11 residue analogue were examined. Single tryptophanyl substitutions at certain positions yielded some active 11-residue analogues. The most effective site for the substitution was the hydrophobic-hydrophilic interface in the amphipathic helical structure. At this position, tryptophan was the most useful amino acid conferring favorable activity to the peptide. The introduced tryptophan played an important anchoring role for the membrane interaction of the peptides. Finally, two 11-residue analogues of GGN5, which exhibited strong bactericidal activity with little hemolytic activity, were obtained as property-optimized candidates for new peptide antibiotic development. Altogether, the present approach not only characterized some important factors for the antimicrobial activity but also provided useful information about peptide engineering to search for potent lead molecules for new peptide antibiotic development. PMID- 14739295 TI - Doxorubicin paradoxically protects cardiomyocytes against iron-mediated toxicity: role of reactive oxygen species and ferritin. AB - The cardiotoxicity induced by the anticancer anthracycline doxorubicin (DOX) is attributed to reactions between iron and reactive oxygen species (ROS) that lead to oxidative damage. We found that DOX forms ROS in H9c2 cardiomyocytes, as shown by dichlorodihydrofluorescein oxidation and the expression of stress-responsive genes such as catalase or aldose reductase. DOX also increased ferritin levels in these cells, particularly the H subunit. A considerable increase in ferritin mRNA levels showed that DOX acted at transcriptional level, but an additional potential mechanism was identified as the down-regulation of iron regulatory protein-2, post-transcriptional inhibitor of ferritin synthesis. Pretreatment with DOX protected H9c2 cells against the damage induced by subsequent exposure to ferric ammonium citrate, and experiments with (55)Fe revealed that the protection was due to the deposition of iron in ferritin. Cytoprotection was also observed when DOX was replaced by glucose/glucose oxidase, a source of H(2)O(2), thus suggesting that DOX increases ferritin synthesis through the action of ROS. This concept was supported by three more lines of evidence. (i) DOX-induced ferritin synthesis was blocked by N-acetylcysteine, a scavenger of ROS. (ii) Mitoxantrone, a ROS-forming analogue, similarly induced ferritin expression and protected the cells against iron toxicity. (iii) 5-Iminodaunorubicin, an analogue lacking ROS-forming activity, did not induce ferritin synthesis or protect the cells against iron toxicity. These results characterize a paradoxically beneficial link between anthracycline-derived ROS, increased ferritin synthesis, and resistance to iron-mediated damage. The role of iron and ROS in anthracycline induced cardiotoxicity may, therefore, be more complex than previously believed. PMID- 14739296 TI - Inactivation of the 25-hydroxyvitamin D 1alpha-hydroxylase and vitamin D receptor demonstrates independent and interdependent effects of calcium and vitamin D on skeletal and mineral homeostasis. AB - We employed a genetic approach to determine whether deficiency of 1,25 dihydroxyvitamin D (1,25(OH)2D) and deficiency of the vitamin D receptor (VDR) produce the same alterations in skeletal and calcium homeostasis and whether calcium can subserve the skeletal functions of 1,25(OH)2D and the VDR. Mice with targeted deletion of the 25-hydroxyvitamin D 1alpha-hydroxylase (1alpha(OH)ase-/ ) gene, the VDR gene, and both genes were exposed to 1) a high calcium intake, which maintained fertility but left mice hypocalcemic; 2) this intake plus three times weekly injections of 1,25(OH)2D3, which normalized calcium in the 1alpha(OH)ase-/- mice only; or 3) a "rescue" diet, which normalized calcium in all mutants. These regimens induced different phenotypic changes, thereby disclosing selective modulation by calcium and the vitamin D system. Parathyroid gland size and the development of the cartilaginous growth plate were each regulated by calcium and by 1,25(OH)2D3 but independent of the VDR. Parathyroid hormone secretion and mineralization of bone reflected ambient calcium levels rather than the 1,25(OH)2D/VDR system. In contrast, increased calcium absorption and optimal osteoblastogenesis and osteoclastogenesis were modulated by the 1,25(OH)2D/VDR system. These studies indicate that the calcium ion and the 1,25(OH)2D/VDR system exert discrete effects on skeletal and calcium homeostasis, which may occur coordinately or independently. PMID- 14739297 TI - Resistance of B16 melanoma cells to CD47-induced negative regulation of motility as a result of aberrant N-glycosylation of SHPS-1. AB - The adhesion receptor SHPS-1 activates the protein-tyrosine-phosphatase SHP-2 and thereby promotes integrin-mediated reorganization of the cytoskeleton. SHPS-1 also contributes to cell-cell communication through association with CD47. Although functional alteration of SHPS-1 is implicated in cellular transformation, the role of the CD47-SHPS-1 interaction in carcinogenesis has been unclear. A soluble SHPS-1 ligand (CD47-Fc) has now been shown to bind to Melan-a non-tumorigenic melanocytes but not to syngeneic B16F10 melanoma cells. Treatment of B16F10 cells with 1-deoxymannojirimycin, which prevents N-glycan processing, restored the ability of SHPS-1 derived from these cells to bind CD47 Fc in vitro, indicating that aberrant N-glycosylation of SHPS-1 impairs CD47 binding in B16F10 cells. CD47-Fc inhibited the migration of Melan-a cells but not that of B16F10 cells. However, a monoclonal antibody that reacts with SHPS-1 on both Melan-a and B16F10 cells inhibited the migration of both cell types similarly. CD47 binding induced proteasome-mediated degradation of SHPS-1 in a tyrosine phosphorylation-independent manner. Furthermore, overexpression of SHPS 1 reduced the level of tyrosine phosphorylation of focal adhesion kinase, and this effect was reversed by CD47 binding. These results suggest that CD47 binds to and thereby down-regulates SHPS-1 on adjacent cells, resulting in inhibition of cell motility. Resistance to this inhibitory mechanism may contribute to the highly metastatic potential of B16 melanoma. PMID- 14739298 TI - A cleaved form of MAGE-A4 binds to Miz-1 and induces apoptosis in human cells. AB - Gankyrin, a recently discovered oncoprotein, is a promising target for drug therapy because it is overexpressed in most hepatocellular carcinomas. Since gankyrin interacts with MAGE-A4, we made several MAGE-A4 mutants and assessed their effects on cell growth. We found that the C-terminal 107 amino acids of MAGE-A4 (MAGE-A4DeltaN1) induced p53-dependent and p53-independent apoptosis. MAGE-A4DeltaN1 increased the p53 protein level, but decreased the p21(Cip1) transcript and protein levels. During apoptosis Bcl-xL was down-regulated and mitochondrial integrity was disrupted. A yeast two-hybrid screen identified Miz-1 as a MAGE-A4DeltaN1-binding protein. MAGE-A4DeltaN1 was recruited through association with Miz-1 to the p21(Cip1) promoter and down-regulated transcription of p21(Cip1). In 293T cells and U-2 OS cells, full-length MAGE-A4 was processed to generate a C-terminal fragment of 104 amino acids with activities similar to MAGE-A4DeltaN1. Processing was inhibited with a broad range caspase inhibitor Z VAD-FMK, but not by site-directed mutagenesis of aspartic acids in MAGE-A4, suggesting an indirect involvement of caspase(s) in the processing. The amount of the processed form was increased by exposure of cells to adriamycin. Transduction with a HIV Tat-MAGE-A4DeltaN1 fusion protein suppressed anchorage-independent growth of gankyrin-overexpressing cells in vitro and in vivo. These results demonstrate that the C-terminal fragment of MAGE-A4 induces apoptosis at least partly by binding to Miz-1, and that the fragment may be exploited as an anticancer agent. Furthermore, the finding that a C-terminal fragment with pro apoptotic activity is generated from full-length MAGE-A4 after genotoxic stress in human cells suggests a novel function for MAGE-A4. PMID- 14739299 TI - A critical intramolecular interaction for protein kinase Cepsilon translocation. AB - Disruption of intramolecular interactions, translocation from one intracellular compartment to another, and binding to isozyme-specific anchoring proteins termed RACKs, accompany protein kinase C (PKC) activation. We hypothesized that in inactive epsilonPKC, the RACK-binding site is engaged in an intramolecular interaction with a sequence resembling its RACK, termed psiepsilonRACK. An amino acid difference between the psiepsilonRACK sequence in epsilonPKC and its homologous sequence in epsilonRACK constitutes a change from a polar non-charged amino acid (asparagine) in epsilonRACK to a polar charged amino acid (aspartate) in epsilonPKC. Here we show that mutating the aspartate to asparagine in epsilonPKC increased intramolecular interaction as indicated by increased resistance to proteolysis, and slower hormone- or PMA-induced translocation in cells. Substituting aspartate for a non-polar amino acid (alanine) resulted in binding to epsilonRACK without activators, in vitro, and increased translocation rate upon activation in cells. Mathematical modeling suggests that translocation is at least a two-step process. Together our data suggest that intramolecular interaction between the psiepsilonRACK site and RACK-binding site within epsilonPKC is critical and rate limiting in the process of PKC translocation. PMID- 14739300 TI - Src kinase activity is essential for osteoclast function. AB - Deletion of the c-src gene impairs osteoclast bone resorbing activity, causing osteopetrosis. Although it has been concluded that restoring only the Src adaptor function at least partly rescues the cell attachment and skeletal phenotypes, the contribution of Src kinase activity remains controversial. Src forms a complex with Pyk2 and Cbl after adhesion-induced stimulation of alpha(V)beta(3) integrin. To demonstrate the importance of the Pyk2-Src association in osteoclasts and to distinguish the contributions of the Src adaptor and kinase activities in cytoskeletal organization and osteoclast function, we expressed mutants of Src and Pyk2 in osteoclasts using adenovirus vectors. Eliminating the Src-binding site on Pyk2 (Pyk2(Y402F)) markedly inhibited bone resorption by osteoclast-like cells, whereas kinase-dead Pyk2 had little effect. Kinase-dead Src, unlike kinase dead Pyk2, markedly inhibited the bone-resorbing activity of wild type osteoclasts and failed to significantly restore bone-resorbing activity to Src(-/ ) osteoclast-like cells. Activation of Src kinase by overexpressing kinase-dead Csk failed to reverse the inhibitory effect of Pyk2(Y402F), suggesting that osteoclastic bone resorption requires both c-Src kinase activity and the targeting of Src kinase by Pyk2. Src-catalyzed phosphorylation of Cbl on Tyr-731 is reported to induce the activation and recruitment of phosphatidylinositol 3 kinase to the cell membrane in a signaling pathway that is critical for osteoclast function. Expressing the Cbl(Y731F) mutant in osteoclasts markedly reduced their bone resorbing activity, suggesting that phosphorylation of Cbl(Y731) and the subsequent recruitment and activation of phosphatidylinositol 3 kinase may be critical signaling events downstream of Src in osteoclasts. PMID- 14739301 TI - The low density lipoprotein receptor-1, LRP1, interacts with the human frizzled-1 (HFz1) and down-regulates the canonical Wnt signaling pathway. AB - Members of the low density lipoprotein receptor family (LDLR), LRP5/6, were shown to interact with the Frizzled (Fz) receptors and to function as Wnt coreceptors. Here we show that mLRP4T100, a minireceptor of LRP1, another member of the LDLR family, interacts with the human Fz-1 (HFz1), previously shown to serve as a receptor transmitting the canonical Wnt-3a-induced signaling cascade. However, in contrast to LRP5/6, mLRP4T100, as well as the full-length LRP1, did not cooperate with HFz1 in transmitting the Wnt-3a signaling but rather repressed it. mLRP4T100 inhibitory effect was displayed also by endocytosis-defective mLRP4T100 mutants, suggesting that LRP1 repressive effect is not attributable to LRP1-mediated enhanced HFz1 internalization and subsequent degradation. Enforced expression of mLRP4T100 decreased the capacity of HFz1 cysteine-rich domain (CRD) to interact with LRP6, in contrast to HFz1-CRD/Wnt-3a interaction that was not disrupted by overexpressing mLRP4T100. These data suggest that LRP1, by sequestering HFz1, disrupts the receptor/coreceptor complex formation, leading to the repression of the canonical Wnt signaling. Thus, this study implies that the ability to interact with Fz receptors is shared by several members of the LDLR family. However, whereas some members of the LDLR family, such as LRP5/6, interact with Fz and serve as Wnt coreceptors, others negatively regulate Wnt signaling, presumably by sequestering Fz. PMID- 14739302 TI - Nerve growth factor withdrawal-mediated apoptosis in naive and differentiated PC12 cells through p53/caspase-3-dependent and -independent pathways. AB - Programmed cell death is regulated in response to a variety of stimuli, including the tumor suppressor protein p53, that can mediate cell cycle arrest through p21/Waf1 and apoptosis through the Bcl-2/Bax equilibrium and caspases. Neuronal cell apoptosis has been reported to require p53, whereas other data suggest that neuronal cell death may be independent of p53. Comparison of wild type PC12 to a temperature-sensitive PC12 cell line that depresses the normal function of p53 has permitted investigation of the importance of p53 in a variety of cell functions. This study examined the role of p53 in trophic factor withdrawal mediated apoptosis in both naive and differentiated PC12 cells. Our data show that as PC12 cells differentiate they are more poised to undergo apoptosis than their undifferentiated counterparts. Survival assays with XTT (sodium 3'-1 (phenylaminocarbonyl)-3,4-tetrazolium-bis(4-methoxy-6-nitro)benzene sulfonic acid) and TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling) demonstrated that lack of p53 is initially protective against apoptosis. The window of protection is about 20 h for naive and 36 h for differentiated cells. Apoptosis involved caspases 3, 6, and 9. However, caspase 3 activation was absent in cells lacking p53, concomitant with the delayed apoptosis. When the expression of caspase 3 was silenced with interference RNA, wild type PC12 cells revealed a morphology and biochemistry similar to PC12[p53ts] cells, indicating that caspase 3 accounts for the observed delay in apoptosis in p53 dysfunction. These results suggest that p53 is important, but not essential, in factor withdrawal-mediated apoptosis. Parallel pathways of caspase-mediated apoptosis are activated later in the absence of functional p53. PMID- 14739303 TI - Mechanisms of the TRIF-induced interferon-stimulated response element and NF kappaB activation and apoptosis pathways. AB - Toll-like receptor-3 is critically involved in host defense against viruses through induction of type I interferons (IFNs). Recent studies suggest that a Toll/interleukin-1 receptor domain-containing adapter protein (TRIF) and two protein kinases (TANK-binding kinase-1 (TBK1) and IkappaB kinase (IKK)-epsilon) are critically involved in Toll-like receptor-3-mediated IFN-beta production through activation of IFN regulatory factor (IRF)-3 and IRF-7. In this study, we demonstrate that TRIF interacts with both IRF-7 and IRF-3. In addition to TBK1 and IKKepsilon, our results indicate that IKKbeta can also phosphorylate IRF-3 and activate the IFN-stimulated response element. TRIF-induced IRF-3 and IRF-7 activation was mediated by TBK1 and its downstream kinases IKKbeta and IKKepsilon. TRIF induced NF-kappaB activation through an IKKbeta- and tumor necrosis factor receptor-associated factor-6-dependent (but not TBK1- and IKKepsilon-dependent) pathway. In addition, TRIF also induced apoptosis through a RIP/FADD/caspase-8-dependent and mitochondrion-independent pathway. Furthermore, our results suggest that the TRIF-induced IFN-stimulated response element and NF kappaB activation and apoptosis pathways are uncoupled and provide a molecular explanation for the divergent effects induced by the adapter protein TRIF. PMID- 14739304 TI - A novel proteomic approach for specific identification of tyrosine kinase substrates using [13C]tyrosine. AB - Proteomic studies to find substrates of tyrosine kinases generally rely on identification of protein bands that are "pulled down" by antiphosphotyrosine antibodies from ligand-stimulated samples. One can obtain erroneous results from such experiments because of two major reasons. First, some proteins might be basally phosphorylated on tyrosine residues in the absence of ligand stimulation. Second, proteins can bind non-specifically to the antibodies or the affinity matrix. Induction of phosphorylation of proteins by ligand must therefore be confirmed by a different approach, which is not always feasible. We have developed a novel proteomic approach to identify substrates of tyrosine kinases in signaling pathways studies based on in vivo labeling of proteins with "light" (12C-labeled) or "heavy" (13C-labeled) tyrosine. This stable isotope labeling in cell culture method enables the unequivocal identification of tyrosine kinase substrates, as peptides derived from true substrates give rise to a unique signature in a mass spectrometry experiment. By using this approach, from a single experiment, we have successfully identified several known substrates of insulin signaling pathway and a novel substrate, polymerase I and transcript release factor, a protein that is implicated in the control of RNA metabolism and regulation of type I collagen promoters. This approach is amenable to high throughput global studies as it simplifies the specific identification of substrates of tyrosine kinases as well as serine/threonine kinases using mass spectrometry. PMID- 14739305 TI - Interaction with leukocytes: phospholipid-stabilized versus albumin-shell microbubbles. AB - PURPOSE: To confirm that BR14 microbubbles (MBs) can be phagocytosed by activated leukocytes, to determine their stability after phagocytosis, and to evaluate how such characteristics influence the fate of neutrophils containing MBs after insonation. MATERIALS AND METHODS: BR14 and human albumin MBs (2 x 10(7)/mL) were incubated with activated human neutrophils (2 x 10(6)/mL) to allow phagocytosis. Deflation rate of the phagocytosed MBs after pulsed insonation (one burst per second for 5 seconds) at 1.8 MHz with peak negative pressure of -540 kPa or 1,340 kPa, lactate dehydrogenase (LDH) leakage, and terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling stain-positive cell count after insonation were compared between the two agents. RESULTS: At -540 kPa, phagocytosed MBs remained nearly unchanged for both agents after insonation. At 1,340 kPa, although human albumin MBs were disrupted on the first or second burst, BR14 MBs remained undisrupted. After -540-kPa insonation, a similar number of apoptotic cells appeared in neutrophils containing human albumin and BR14 MBs. At -540 kPa, LDH leakage was limited in human albumin MBs and BR14 MBs. At -1,340 kPa, LDH leakage was significantly increased in human albumin MBs and BR14 MBs (P <.01, both vs -540 kPa). Apoptotic cells were significantly decreased in human albumin MBs and BR14 MBs (P <.01, both vs -540 kPa). LDH leakage was lower and apoptotic cell count was greater in BR14 MB-containing neutrophils than in human albumin MB-containing neutrophils (both P <.01). CONCLUSION: Compared with human albumin MBs, BR14 MBs were more stable after phagocytosis with insonation. This stability is associated with less disruption and greater induction of apoptosis in leukocytes after relatively high-pressure insonation in the range for diagnostic use. PMID- 14739306 TI - T2-weighted MR imaging in the assessment of cirrhotic liver. AB - PURPOSE: To assess if T2-weighted magnetic resonance (MR) imaging provides added diagnostic value in combination with dynamic gadolinium-enhanced MR imaging in the detection and characterization of nodular lesions in cirrhotic liver. MATERIALS AND METHODS: Two readers retrospectively and independently analyzed 54 MR imaging studies in 52 patients with cirrhosis. In session 1, readers reviewed T1-weighted and dynamic gadolinium-enhanced images. In session 2, readers reviewed T1-weighted, dynamic gadolinium-enhanced, and respiratory-triggered T2 weighted fast spin-echo images. Readers identified and characterized all focal lesions by using a scale of 1-4 (1, definitely benign; 4, definitely malignant). Multireader correlated receiver operating characteristic (ROC) analysis was employed to assess radiologist performance in session 2 compared with session 1. The difference in the areas under the ROC curves for the two sessions was tested. In a third session, readers assessed conspicuity of biopsy-proved lesions on T2 weighted MR images by using a scale of 1-3 (1, not seen; 3, well seen) and identified causes of reduced conspicuity. RESULTS: Two additional benign lesions were detected by each reader in session 2. Fifty-five lesions had pathologic verification, including 32 malignant, three high-grade dysplastic, and 20 benign nodules. There was no significant difference in the area under the ROC curves between the two sessions (P =.48). Thirty-two lesions were inconspicuous on T2 weighted MR images because of parenchymal heterogeneity, breathing artifacts (particularly in patients with ascites), and lesion isointensity with liver parenchyma. T2-weighted MR imaging was useful in the evaluation of cysts and lymph nodes. CONCLUSION: T2-weighted MR imaging does not provide added diagnostic value in the detection and characterization of focal lesions in cirrhotic liver. PMID- 14739307 TI - Myocardial tissue tracking with two-dimensional cine displacement-encoded MR imaging: development and initial evaluation. AB - A breath-hold two-dimensional cine magnetic resonance (MR) pulse sequence based on displacement encoding with stimulated echoes (DENSE) for quantitative myocardial motion tracking was developed and evaluated. In the sequence, complementary spatial modulation of magnetization was used for time-independent artifact suppression, and echo-planar imaging was used for rapid data sampling. Twelve healthy volunteers underwent cine DENSE MR imaging, and six of them also underwent conventional MR imaging myocardial tagging. The circumferential shortening component of strain (E(cc)) was measured on cine DENSE MR images and conventional tagged MR images. With complementary spatial modulation of magnetization, 10% or less of the total cine DENSE MR image energy was attributed to an artifact-generating echo during systolic imaging. Two-dimensional intramyocardial displacement and strain were measured at cine DENSE MR imaging with spatial resolution and temporal resolution of 2.7 x 2.7 mm and 60 msec, respectively. E(cc) measured at cine DENSE MR imaging correlated well with that measured at conventional MR imaging tagging (slope = 0.88, intercept = 0.00, R = 0.87). PMID- 14739308 TI - Patellar articular cartilage lesions: in vitro MR imaging evaluation after placement in gadopentetate dimeglumine solution. AB - PURPOSE: To evaluate T1-weighted magnetic resonance (MR) imaging after diffusion of gadopentetate dimeglumine for visualization of articular cartilage lesions. MATERIALS AND METHODS: MR imaging was performed in eight human cadaveric patella specimens immediately and 4 hours after placement into a vessel filled with gadopentetate dimeglumine solution (2.5 mmol/L). T1-weighted spin-echo and inversion-recovery turbo spin-echo MR sequences with nulled cartilage signal (inversion time of 300 msec) were used. In a total of 128 articular cartilage areas, MR imaging findings were compared with macroscopic and histopathologic findings. Pathologic evaluation was performed by one musculoskeletal pathologist. With knowledge of pathologic observations, MR images were analyzed by one musculoskeletal radiologist with regard to intrinsic signal intensity characteristics and surface abnormalities of articular cartilage. RESULTS: Histopathologic findings demonstrated 67 areas of normal articular cartilage and 66 cartilage lesions (grade 1, n = 19; grade 2, n = 15; grade 3, n = 26; grade 4, n = 6). All grade 3 and 4 lesions could be identified on MR images obtained immediately after submersion and after 4 hours. Ninety-four percent of grade 1 and 2 lesions were identified as areas of predominantly decreased contrast enhancement on delayed MR images obtained with both sequences. MR images obtained immediately after submersion demonstrated abnormal signal intensity in only 9% and 12% of grade 1 and 2 lesions, respectively. CONCLUSION: T1-weighted MR images obtained in vitro after gadopentetate dimeglumine diffusion allow demonstration of articular cartilage surface lesions and early stages of cartilage degradation. PMID- 14739309 TI - Value of transcatheter arterial embolization with coils and n-butyl cyanoacrylate for long-term hepatic arterial infusion chemotherapy. AB - PURPOSE: To assess the value of transcatheter arterial embolization (TAE) of splanchnic arterial branches to allow continuous application of repeat hepatic arterial infusion chemotherapy (HAIC). MATERIALS AND METHODS: In 128 patients with unresectable advanced liver cancer, percutaneous implantation of a port catheter system and TAE of splanchnic arteries with coils and/or n-butyl cyanoacrylate (NBCA) were performed. Parameters included (a) methods selected for catheter placement; (b) embolic materials used (coils and/or NBCA, number of coils, administration rate of NBCA-iodized oil) for TAE of splanchnic arteries, details of embolized arteries, and frequency of recanalization; (c) ability to prevent gastrointestinal symptoms by avoiding inflow of anticancer drugs into extrahepatic adjacent organs and to maintain distribution of contrast agents in liver, as well as management of difficulties encountered; (d) complications related to catheter system implantation or to long-term HAIC and management of such complications; and (e) final success in performing scheduled HAIC while maintaining distribution over liver via a single route without gastrointestinal symptoms caused by inflow of anticancer drugs. Fisher exact test was used to compare recanalization rate between coil-embolized and NBCA- or NBCA-coil embolized arteries, and frequency of heterogeneously poor distribution was compared between patients with single and those with multiple hepatic arteries. RESULTS: Embolization was successful during first catheterization in 326 arteries and during follow-up in 10. In 119 (93.0%) of 128 patients, repeat HAIC was effective until death or the time of this writing (observation period, 2-47 months). HAIC was continued in two patients, although anticancer drugs did not distribute to all liver tumors. Arteries once embolized with coils alone spontaneously recanalized at a significantly higher rate than those with NBCA (eight of 192 vs one of 144, P =.048). Rate of heterogeneously poor distribution was significantly higher in those with two or more hepatic arteries than in those with one (seven of 17 vs nine of 111, P =.001). CONCLUSION: TAE for various splanchnic organs is useful for efficient performance of long-term HAIC. PMID- 14739310 TI - Bjork-Shiley convexoconcave valves: susceptibility artifacts at brain MR imaging and mechanical valve fractures. AB - PURPOSE: To assess the relationship between heart valve history and susceptibility artifacts at magnetic resonance (MR) imaging of the brain in patients with Bjork-Shiley convexoconcave (BSCC) valves. MATERIALS AND METHODS: MR images of the brain were obtained in 58 patients with prosthetic heart valves: 20 patients had BSCC valve replacements, and 38 had other types of heart valves. Two experienced neuroradiologists determined the presence or absence of susceptibility artifacts in a consensus reading. Artifacts were defined as characteristic black spots that were visible on T2*-weighted gradient-echo MR images. The statuses of the 20 explanted BSCC valves-specifically, whether they were intact or had an outlet strut fracture (OSF) or a single-leg fracture (SLF) had been determined earlier. Number of artifacts seen at brain MR imaging was correlated with explanted valve status, and differences were analyzed with nonparametric statistical tests. RESULTS: Significantly more patients with BSCC valves (17 [85%] of 20 patients) than patients with other types of prosthetic valves (18 [47%] of 38 patients) had susceptibility artifacts at MR imaging (P =.005). BSCC valve OSFs were associated with a significantly higher number of artifacts than were intact BSCC valves (P =.01). No significant relationship between SLF and number of artifacts was observed. CONCLUSION: Susceptibility artifacts at brain MR imaging are not restricted to patients with BSCC valves. These artifacts can be seen on images obtained in patients with various other types of fractured and intact prosthetic heart valves. PMID- 14739311 TI - Colorectal polyps and cancers in asymptomatic average-risk patients: evaluation with CT colonography. AB - PURPOSE: To compare thin-section multi-detector row computed tomographic (CT) colonography with conventional colonoscopy in the evaluation of colorectal polyps and cancer in asymptomatic average-risk patients. MATERIALS AND METHODS: Sixty eight asymptomatic men (age > 50 years) scheduled to undergo screening colonoscopy were enrolled in this study. CT colonography was followed by conventional colonoscopy, performed on the same day. Supine and prone CT colonography were performed after colonic insufflation with room air. A gastroenterologist measured all polyps, which were categorized as 1-5, 6-9, or over 10 mm. Biopsy and histologic evaluation were performed of all polyps. CT colonography and colonoscopy results were compared for location, size, and morphology of detected lesions. Point estimates and 95% CIs were provided for specificity and sensitivity of CT by using results at conventional colonoscopy as the reference standard. RESULTS: At colonoscopy, 98 polyps were identified in 39 patients; 21 (21.4%) of 98 were detected at CT colonography. Sensitivity was 11.5% (nine of 78) for polyps 1-5 mm, 52.9% (nine of 17) for polyps 6-9 mm, and 100% (three of three) for polyps over 10 mm. Results at colonoscopy were normal in 29 (42.6%) of 68 patients; at CT colonography, results were correctly identified as normal in 26 of these 29 patients. In one of these patients, a lesion larger than 10 mm was detected at CT colonography. The per-patient specificity of CT was 89.7% (26 of 29; 95% CI: 72.7%, 97.8%). The mean time for CT image interpretation was 9 minutes. CONCLUSION: In patients at average risk for colorectal cancer, CT colonography is a sensitive and specific screening test for detecting polyps 10 mm or larger; the sensitivity for detecting smaller polyps is decreased. Examination findings can be interpreted in a clinically feasible amount of time. PMID- 14739312 TI - Strategies for CT radiation dose optimization. AB - Recent technologic advances have markedly enhanced the clinical applications of computed tomography (CT). While the benefits of CT exceed the harmful effects of radiation exposure in patients, increasing radiation doses to the population have raised a compelling case for reduction of radiation exposure from CT. Strategies for radiation dose reduction are difficult to devise, however, because of a lack of guidelines regarding CT examination and scanning techniques. Various methods and strategies based on individual patient attributes and CT technology have been explored for dose optimization. It is the purpose of this review article to outline basic principles of CT radiation exposure and emphasize the need for CT radiation dose optimization based on modification of scanning parameters and application of recent technologic innovations. PMID- 14739313 TI - Low-grade gliomas and focal cortical developmental malformations: differentiation with proton MR spectroscopy. AB - PURPOSE: To assess proton magnetic resonance (MR) spectroscopy in differentiating between low-grade gliomas and focal cortical developmental malformations (FCDMs). MATERIALS AND METHODS: Eighteen patients with seizures and a cortical brain lesion on MR images were studied with proton MR spectroscopy. A metabolite ratio analysis was performed, and the metabolite signals in the lesion core were compared with those in the contralateral centrum semiovale and in the corresponding brain sites in 18 control subjects to separately obtain the changes in N-acetylaspartate (NAA), choline-containing compounds (Cho), and creatine phosphocreatine (Cr). Ten patients had a low-grade glioma (three, oligodendrogliomas; three, oligoastrocytomas; three, astrocytomas; and one, pilocytic astrocytoma), and eight had FCDM (five, focal cortical dysplasias and three, dysembryoplastic neuroepithelial tumors). Linear discriminant analysis and Student t test were used for statistical comparisons. RESULTS: Loss of NAA and increase of Cho were more pronounced in low-grade gliomas than in FCDMs (NAA, 72% +/- 15 [+/- SD] vs -29% +/- 22, P <.001; Cho, 117% +/- 56 vs 21% +/- 66, P <.01). Changes in NAA and Cho helped differentiate low-grade gliomas from FCDMs, and changes in Cho and Cr helped differentiate astrocytomas from oligodendrogliomas and oligoastrocytomas. Metabolite NAA/Cho and NAA/Cr ratios helped differentiate low-grade gliomas from FCDMs but did not differentiate glioma subtypes. CONCLUSION: MR spectroscopy allows distinction between low-grade gliomas and FCDMs and between low-grade glioma subtypes. Metabolite changes are more informative than are metabolite ratios. PMID- 14739314 TI - CT pulmonary angiography: quantification of pulmonary embolus as a predictor of patient outcome--initial experience. AB - PURPOSE: To determine whether quantification of pulmonary embolus (PE) with computed tomographic (CT) pulmonary angiography by using a standardized index is a predictor of patient outcome. MATERIALS AND METHODS: Multi-detector row CT was performed in 59 hospitalized patients (mean age, 61 years; age range, 22-89 years). PE was identified retrospectively by two radiologists who were blinded to patient outcome. A pulmonary arterial obstruction index was derived for each set of images on the basis of embolus size and location. By using logistic regression, PE indexes were compared with patient outcome-survival or death-to determine if there was a correlation between PE volume and survival. RESULTS: The PE index is a significant predictor of patient outcome (P =.002). One of 53 patients (1.9%) with an index of less than 60% died. Cause of death was end-stage malignancy. Five of six patients (83%) with an index of 60% and higher died. All five deaths were related to the presence of PE. The one survivor with a PE index higher than 60% received thrombolytic therapy. By using a cutoff of 60%, the PE index was used to identify 52 of 53 (98%) patients who survived and five of six (83%) patients who died. CONCLUSION: Preliminary evidence suggests that quantification of clot with CT pulmonary angiography is an important predictor of patient death in the setting of PE. PMID- 14739315 TI - Computer-aided classification of BI-RADS category 3 breast lesions. AB - PURPOSE: To evaluate a system for computer-aided classification (CAC) of lesions assigned to Breast Imaging Reporting and Data System (BI-RADS) category 3 at conventional mammographic interpretation. MATERIALS AND METHODS: A CAC system was used to analyze 106 cases of lesions (42 malignant) that at blinded retrospective interpretation were assigned to BI-RADS category 3 by at least two of four radiologists. The CAC system automatically extracted from the digitized mammograms quantitative features that characterized the lesions. The system then used a classification scheme to score the lesions by the likelihood of their malignancy on the basis of these features. The classification scheme was trained with 646 pathologically proved cases (323 malignant), and the results were tested with receiver operating characteristic (ROC) analysis by using the jackknife method. Sensitivity, specificity, positive predictive value, and accuracy were calculated. Category 3 lesions were stratified among BI-RADS categories 2-5 according to CAC-assigned lesion score, and this classification was compared with the results of pathologic analysis. RESULTS: Jackknife analysis of CAC results in the training data set yielded a sensitivity of 94%, specificity of 78%, positive predictive value of 81%, and area under the ROC curve of 0.90. Of the 42 malignant lesions that had been classified at conventional interpretation as probably benign, nine were assigned by the CAC system to BI-RADS category 4, and 29 were assigned to category 5. The CAC system correctly upgraded the BI-RADS classification of these 38 lesions (sensitivity, 90%) and incorrectly upgraded the classification of only 20 benign lesions (specificity, 69%). CONCLUSION: The CAC system scored 38 of the 42 malignant lesions initially assigned to BI-RADS category 3 as BI-RADS category 4 or 5, and thus correctly upgraded the category in 90% of these lesions. PMID- 14739316 TI - Variations in pulmonary venous drainage to the left atrium: implications for radiofrequency ablation. AB - PURPOSE: To evaluate and classify the various drainage patterns of the pulmonary veins as depicted with thin-section chest computed tomography (CT). MATERIALS AND METHODS: Thin-section (2.5-mm collimation) contrast material-enhanced CT scans of 201 consecutive patients obtained over a 3-month period for diagnosis of pulmonary embolism (n = 197), pulmonary vein stenosis (n = 2), or aortic injury (n = 2) were routinely reviewed in transverse and (if necessary) coronal and coronal-oblique imaging planes. A classification was formulated based on both the number of venous ostia on each side and the drainage patterns of pulmonary veins. The frequency of each pattern was determined, and association with atrial arrhythmia was assessed with the chi(2) and Fisher exact tests. RESULTS: Most patients (n = 142, 71%) had two ostia on the right side for upper and lower lobe veins. Fifty-six patients (28%) had three to five ostia on the right side, which were due to one or two separate middle lobe vein ostia in 52 (26%) patients. Three patients (2%) had a single venous ostium on the right side. Most patients (n = 173, 86%) had two ostia on the left side for upper and lower lobe veins. The remainder (n = 28, 14%) had a single ostium. There was no significant association between any particular venous drainage pattern and atrial arrhythmia; however, patients with a separate ostia for the right middle lobe pulmonary vein(s) tended to have a higher frequency of atrial arrhythmia than those with other patterns (P =.053). CONCLUSION: A classification system to succinctly describe pulmonary venous drainage patterns was developed. Right-sided venous drainage was more variable than left-sided venous drainage. One-quarter of patients had more than two venous ostia on the right side. PMID- 14739317 TI - Myoglobin cavities provide interior ligand pathway. AB - The myoglobin protein binds oxygen and catalyzes NO oxidation. As a key model protein, its dynamics have been well studied by spectroscopy and by crystallography as well as by simulation. Nonetheless, visualization of the mechanism of movement of ligands within myoglobin has been difficult. Coordinates of the A1 and A3 taxonomic spectral states of myoglobin from the 1 A crystal structure (1a6g) are generated as consistent sets of correlated clusters of residues with A or B crystal alternates. Analysis of cavities in these A1 and A3 conformations clarifies the pathway of ligand motion from distal entry through interior movement to the proximal side of the heme. Cavities opened up by buried alternate conformations link the distal to the proximal side of the heme. Structural conservation highlights the relevance of this pathway to human neuroglobin. Cavity migration via myoglobin crystal alternates provides a specific link of protein structure to protein dynamics and protein function and demonstrates the relevance of substates (discrete disorder) to function for all proteins. PMID- 14739318 TI - Disabling the folding catalyst is the last critical step in alpha-lytic protease folding. AB - Alpha-Lytic protease (alphaLP) is an extracellular bacterial pro-protease marked by extraordinary conformational rigidity and a highly cooperative barrier to unfolding. Although these properties successfully limit its proteolytic destruction, thereby extending the functional lifetime of the protease, they come at the expense of foldability (t(1/2) = 1800 yr) and thermodynamic stability (native alphaLP is less stable than the unfolded species). Efficient folding has required the coevolution of a large N-terminal pro region (Pro) that rapidly catalyzes alphaLP folding (t(1/2) = 23 sec) and shifts the thermodynamic equilibrium in favor of folded protease through tight native-state binding. Release of active alphaLP from this stabilizing, but strongly inhibitory, complex requires the proteolytic destruction of Pro. alphaLP is capable of initiating Pro degradation via cleavage of a flexible loop within the Pro C-terminal domain. This single cleavage event abolishes Pro catalysis while maintaining strong native-state binding. Thus, the loop acts as an Achilles' heel by which the Pro foldase machinery can be safely dismantled, preventing Pro-catalyzed unfolding, without compromising alphaLP native-state stability. Once the loop is cleaved, Pro is rapidly degraded, releasing active alphaLP. PMID- 14739319 TI - Thermal stability of human alpha-crystallins sensed by amide hydrogen exchange. AB - The alpha-crystallins, alphaA and alphaB, are major lens structural proteins with chaperone-like activity and sequence homology to small heat-shock proteins. As yet, their crystal structures have not been determined because of the large size and heterogeneity of the assemblies they form in solution. Because alpha crystallin chaperone activity increases with temperature, understanding structural changes of alpha-crystallin as it is heated may help elucidate the mechanism of chaperone activity. Although a variety of techniques have been used to probe changes in heat-stressed alpha-crystallin, the results have not yet yielded a clear understanding of chaperone activity. We report examination of native assemblies of human lens alpha-crystallin using hydrogen/deuterium exchange in conjunction with enzymatic digestion and analysis by mass spectrometry. This technique has the advantage of sensing structural changes along much of the protein backbone and being able to detect changes specific to alphaA and alphaB in the native assembly. The reactivity of the amide linkages to hydrogen/deuterium exchange was determined for 92% of the sequence of alphaA and 99% of alphaB. The behavior of alphaA and alphaB is remarkably similar. At low temperatures, there are regions at the beginning of the alpha-crystallin domains in both alphaA and alphaB that have high protection to isotope exchange, whereas the C termini offer little protection. The N terminus of alphaA also has low protection. With increasing temperatures, both proteins show gradual unfolding. The maximum percent change in exposure with increasing temperatures was found in alphaA 72-75 and alphaB 76-79, two regions considered critical for chaperone activity. PMID- 14739320 TI - High-temperature solution NMR structure of TmCsp. AB - Cold shock proteins (Csps) are assumed to play a central role in the regulation of gene expression under cold shock conditions. Acting as single-stranded nucleic acid-binding proteins, they trigger the translation process and are therefore involved in the compensation of the influence of low temperatures (cold shock) upon the cell metabolism. However, it is unknown so far how Csps are switched on and off as a function of temperature. The aim of the present study is the study of possible structural changes responsible for this switching process. (1)H-(15)N HSQC spectra recorded at different temperatures and chemical-shift analysis have indicated subtle conformational changes for the cold-shock protein from the hyperthermophilic bacterium Thermotoga maritima (TmCsp) when the temperature is elevated from 303 K to its physiological temperature (343 K). The three dimensional structure of TmCsp was determined by nuclear magnetic resonance (NMR) spectroscopy at 343 K to obtain quantitative information concerning these structural changes. By use of residual dipolar couplings, the loss of NOE information at high temperature could be compensated successfully. Most pronounced conformational changes compared with room-temperature conditions are observed for amino acid residues closely neighbored to two characteristic beta bulges and a well-defined loop region of the protein. Because the residues shown to be responsible for the interaction of TmCsp with single-stranded nucleic acids can almost exclusively be found within these regions, nucleic acid-binding activity might be down-regulated with increasing temperature by the described conformational changes. PMID- 14739321 TI - Analysis of forces that determine helix formation in alpha-proteins. AB - A model for prediction of alpha-helical regions in amino acid sequences has been tested on the mainly-alpha protein structure class. The modeling represents the construction of a continuous hypothetical alpha-helical conformation for the whole protein chain, and was performed using molecular mechanics tools. The positive prediction of alpha-helical and non-alpha-helical pentapeptide fragments of the proteins is 79%. The model considers only local interactions in the polypeptide chain without the influence of the tertiary structure. It was shown that the local interaction defines the alpha-helical conformation for 85% of the native alpha-helical regions. The relative energy contributions to the energy of the model were analyzed with the finding that the van der Waals component determines the formation of alpha-helices. Hydrogen bonds remain at constant energy independently whether alpha-helix or non-alpha-helix occurs in the native protein, and do not determine the location of helical regions. In contrast to existing methods, this approach additionally permits the prediction of conformations of side chains. The model suggests the correct values for ~60% of all chi-angles of alpha-helical residues. PMID- 14739322 TI - Solvent-amino acid interaction energies in three-dimensional-lattice Monte Carlo simulations of a model 27-mer protein: Folding thermodynamics and kinetics. AB - Amino acid residue-solvent interactions are required for lattice Monte Carlo simulations of model proteins in water. In this study, we propose an interaction energy scale that is based on the interaction scale by Miyazawa and Jernigan. It permits systematic variation of the amino acid-solvent interactions by introducing a contrast parameter for the hydrophobicity, C(s), and a mean attraction parameter for the amino acids, omega. Changes in the interaction energies strongly affect many protein properties. We present an optimized energy parameter set for best representing realistic behavior typical for many proteins (fast folding and high cooperativity for single chains). Our optimal parameters feature a much weaker hydrophobicity contrast and mean attraction than does the original interaction scale. The proposed interaction scale is designed for calculating the behavior of proteins in bulk and at interfaces as a function of solvent characteristics, as well as protein size and sequence. PMID- 14739323 TI - An automated in vitro protein folding screen applied to a human dynactin subunit. AB - The preparation of proteins for structural and functional analysis using the Escherichia coli expression system is often hampered by the formation of insoluble intracellular protein aggregates (inclusion bodies). Transferring those proteins into their native states by in vitro protein folding requires screening for the best buffer conditions and suitable additives. However, it is difficult to assess the success of such a screen if no biological assay is available. We established a fully automated folding screen and a system to detect folded protein that is based on analytical hydrophobic interaction chromatography and tryptophan fluorescence spectroscopy. The system was evaluated with two model enzymes (carbonic anhydrase II and malate dehydrogenase), and was successfully applied to the folding of the p22 subunit of human dynactin, which is expressed in inclusion bodies in E. coli. The described screen allows for high-throughput folding analysis of inclusion body proteins for structural and functional analyses. PMID- 14739324 TI - Accurate and efficient loop selections by the DFIRE-based all-atom statistical potential. AB - The conformations of loops are determined by the water-mediated interactions between amino acid residues. Energy functions that describe the interactions can be derived either from physical principles (physical-based energy function) or statistical analysis of known protein structures (knowledge-based statistical potentials). It is commonly believed that statistical potentials are appropriate for coarse-grained representation of proteins but are not as accurate as physical based potentials when atomic resolution is required. Several recent applications of physical-based energy functions to loop selections appear to support this view. In this article, we apply a recently developed DFIRE-based statistical potential to three different loop decoy sets (RAPPER, Jacobson, and Forrest-Woolf sets). Together with a rotamer library for side-chain optimization, the performance of DFIRE-based potential in the RAPPER decoy set (385 loop targets) is comparable to that of AMBER/GBSA for short loops (two to eight residues). The DFIRE is more accurate for longer loops (9 to 12 residues). Similar trend is observed when comparing DFIRE with another physical-based OPLS/SGB-NP energy function in the large Jacobson decoy set (788 loop targets). In the Forrest-Woolf decoy set for the loops of membrane proteins, the DFIRE potential performs substantially better than the combination of the CHARMM force field with several solvation models. The results suggest that a single-term DFIRE-statistical energy function can provide an accurate loop prediction at a fraction of computing cost required for more complicate physical-based energy functions. A Web server for academic users is established for loop selection at the softwares/services section of the Web site http://theory.med.buffalo.edu/. PMID- 14739325 TI - An accurate, residue-level, pair potential of mean force for folding and binding based on the distance-scaled, ideal-gas reference state. AB - Structure prediction on a genomic scale requires a simplified energy function that can efficiently sample the conformational space of polypeptide chains. A good energy function at minimum should discriminate native structures against decoys. Here, we show that a recently developed, residue-specific, all-atom knowledge-based potential (167 atomic types) based on distance-scaled, finite ideal-gas reference state (DFIRE-all-atom) can be substantially simplified to 20 residue types located at side-chain center of mass (DFIRE-SCM) without a significant change in its capability of structure discrimination. Using 96 standard multiple decoy sets, we show that there is only a small reduction (from 80% to 78%) in success rate of ranking native structures as the top 1. The success rate is higher than two previously developed, all-atom distance-dependent statistical pair potentials. Applied to structure selections of 21 docking decoys without modification, the DFIRE-SCM potential is 29% more successful in recognizing native complex structures than an all-atom statistical potential trained by a database of dimeric interfaces. The potential also achieves 92% accuracy in distinguishing true dimeric interfaces from artificial crystal interfaces. In addition, the DFIRE potential with the C(alpha) positions as the interaction centers recognizes 123 native structures out of a comprehensive 125 protein TOUCHSTONE decoy set in which each protein has 24,000 decoys with only C(alpha) positions. Furthermore, the performance by DFIRE-SCM on newly established 25 monomeric and 31 docking Rosetta-decoy sets is comparable to (or better than in the case of monomeric decoy sets) that of a recently developed, all-atom Rosetta energy function enhanced with an orientation-dependent hydrogen bonding potential. PMID- 14739326 TI - Phylogenetic and evolutionary analysis of the PLUNC gene family. AB - The PLUNC family of human proteins are candidate host defense proteins expressed in the upper airways. The family subdivides into short (SPLUNC) and long (LPLUNC) proteins, which contain domains predicted to be structurally similar to one or both of the domains of bactericidal/permeability-increasing protein (BPI), respectively. In this article we use analysis of the human, mouse, and rat genomes and other sequence data to examine the relationships between the PLUNC family proteins from humans and other species, and between these proteins and members of the BPI family. We show that PLUNC family clusters exist in the mouse and rat, with the most significant diversification in the locus occurring for the short PLUNC family proteins. Clear orthologous relationships are established for the majority of the proteins, and ambiguities are identified. Completion of the prediction of the LPLUNC4 proteins reveals that these proteins contain approximately a 150-residue insertion encoded by an additional exon. This insertion, which is predicted to be largely unstructured, replaces the structure homologous to the 40s hairpin of BPI. We show that the exon encoding this region is anomalously variable in size across the LPLUNC proteins, suggesting that this region is key to functional specificity. We further show that the mouse and human PLUNC family orthologs are evolving rapidly, which supports the hypothesis that these proteins are involved in host defense. Intriguingly, this rapid evolution between the human and mouse sequences is replaced by intense purifying selection in a large portion of the N-terminal domain of LPLUNC4. Our data provide a basis for future functional studies of this novel protein family. PMID- 14739328 TI - Automated selection of positions determining functional specificity of proteins by comparative analysis of orthologous groups in protein families. AB - The increasing volume of genomic data opens new possibilities for analysis of protein function. We introduce a method for automated selection of residues that determine the functional specificity of proteins with a common general function (the specificity-determining positions [SDP] prediction method). Such residues are assumed to be conserved within groups of orthologs (that may be assumed to have the same specificity) and to vary between paralogs. Thus, considering a multiple sequence alignment of a protein family divided into orthologous groups, one can select positions where the distribution of amino acids correlates with this division. Unlike previously published techniques, the introduced method directly takes into account nonuniformity of amino acid substitution frequencies. In addition, it does not require setting arbitrary thresholds. Instead, a formal procedure for threshold selection using the Bernoulli estimator is implemented. We tested the SDP prediction method on the LacI family of bacterial transcription factors and a sample of bacterial water and glycerol transporters belonging to the major intrinsic protein (MIP) family. In both cases, the comparison with available experimental and structural data strongly supported our predictions. PMID- 14739327 TI - Zymogenic and enzymatic properties of the 70-80 loop mutants of factor X/Xa. AB - The Ca(2+) binding 70-80 loop of factor X (fX) contains one basic (Arg(71)) and three acidic (Glu(74), Glu(76), and Glu(77)) residues whose contributions to the zymogenic and enzymatic properties of the protein have not been evaluated. We prepared four Ala substitution mutants of fX (R71A, E74A, E76A, and E77A) and characterized their activation kinetics by the factor VIIa and factor IXa in both the absence and presence of cofactors. Factor VIIa exhibited normal activity toward E74A and E76A and less than a twofold impaired activity toward R71A and E77A in both the absence and presence of tissue factor. Similarly, factor IXa in the absence of factor VIIIa exhibited normal activity toward both E74A and E76A; however, its activity toward R71A and E77A was impaired approximately two- to threefold. In the presence of factor VIIIa, factor IX activated all mutants with approximately two- to fivefold impaired catalytic efficiency. In contrast to changes in their zymogenic properties, all mutant enzymes exhibited normal affinities for factor Va, and catalyzed the conversion of prothrombin to thrombin with normal catalytic efficiencies. However, further studies revealed that the affinity of mutant enzymes for interaction with metal ions Na(+) and Ca(2+) was impaired. These results suggest that although charged residues of the 70-80 loop play an insignificant role in fX recognition by the factor VIIa-tissue factor complex, they are critical for the substrate recognition by factor IXa in the intrinsic Xase complex. The results further suggest that mutant residues do not play a specific role in the catalytic function of fXa in the prothrombinase complex. PMID- 14739329 TI - Role of Phe283 in enzymatic reaction of cyclodextrin glycosyltransferase from alkalophilic Bacillus sp.1011: Substrate binding and arrangement of the catalytic site. AB - Cyclodextrin glycosyltransferase (CGTase) belonging to the alpha-amylase family mainly catalyzes transglycosylation and produces cyclodextrins from starch and related alpha-1,4-glucans. The catalytic site of CGTase specifically conserves four aromatic residues, Phe183, Tyr195, Phe259, and Phe283, which are not found in alpha-amylase. To elucidate the structural role of Phe283, we determined the crystal structures of native and acarbose-complexed mutant CGTases in which Phe283 was replaced with leucine (F283L) or tyrosine (F283Y). The temperature factors of the region 259-269 in native F283L increased >10 A(2) compared with the wild type. The complex formation with acarbose not only increased the temperature factors (>10 A(2)) but also changed the structure of the region 257 267. This region is stabilized by interactions of Phe283 with Phe259 and Leu260 and plays an important role in the cyclodextrin binding. The conformation of the side-chains of Glu257, Phe259, His327, and Asp328 in the catalytic site was altered by the mutation of Phe283 with leucine, and this indicates that Phe283 partly arranges the structure of the catalytic site through contacts with Glu257 and Phe259. The replacement of Phe283 with tyrosine decreased the enzymatic activity in the basic pH range. The hydroxyl group of Tyr283 forms hydrogen bonds with the carboxyl group of Glu257, and the pK(a) of Glu257 in F283Y may be lower than that in the wild type. PMID- 14739331 TI - A simple formalism on dynamics of proteins on potential energy landscapes. AB - We present a simple formalism for the dynamics of proteins on a potential energy landscape, using connectedness of configurational domains as an order parameter. This formalism clearly shows that the energy bias required to form a unit correct contact toward the native configuration of a two-state folder, to overcome Levinthal's paradox, is E(bias) congruent with RT ln 2. This result agrees well with earlier studies and indicates that the bias is mainly due to hydrophobic interaction. Further investigations have shown that the landscape funnel could be experimentally mapped onto a two-dimensional space formed by denaturant concentration and the connectedness of configurational domains. The theoretical value of the depth-of-folding funnel in terms of denaturant concentration has been calculated for a model protein (P450cam), which agrees well with the experimental value. Using our model, it is also possible to explain the turnover nature of heat-capacity change upon unfolding of proteins and the existence of enthalpy and entropy convergence temperatures during unfolding without any strict assumptions as proposed in earlier studies. PMID- 14739330 TI - Hydrogen/deuterium exchange studies of native rabbit MM-CK dynamics. AB - Creatine kinase (CK) isoenzymes catalyse the reversible transfer of a phosphoryl group from ATP onto creatine. This reaction plays a very important role in the regulation of intracellular ATP concentrations in excitable tissues. CK isoenzymes are highly resistant to proteases in native conditions. To appreciate localized backbone dynamics, kinetics of amide hydrogen exchange with deuterium was measured by pulse-labeling the dimeric cytosolic muscle CK isoenzyme. Upon exchange, the protein was digested with pepsin, and the deuterium content of the resulting peptides was determined by liquid chromatography coupled to mass spectrometry (MS). The deuteration kinetics of 47 peptides identified by MS/MS and covering 96% of the CK backbone were analyzed. Four deuteration patterns have been recognized: The less deuterated peptides are located in the saddle-shaped core of CK, whereas most of the highly deuterated peptides are close to the surface and located around the entrance to the active site. Their exchange kinetics are discussed by comparison with the known secondary and tertiary structures of CK with the goal to reveal the conformational dynamics of the protein. Some of the observed dynamic motions may be linked to the conformational changes associated with substrate binding and catalytic mechanism. PMID- 14739332 TI - The role of hydrogen bonding in the enzymatic reaction catalyzed by HIV-1 protease. AB - The hydrogen-bond network in various stages of the enzymatic reaction catalyzed by HIV-1 protease was studied through quantum-classical molecular dynamics simulations. The approximate valence bond method was applied to the active site atoms participating directly in the rearrangement of chemical bonds. The rest of the protein with explicit solvent was treated with a classical molecular mechanics model. Two possible mechanisms were studied, general-acid/general-base (GA/GB) with Asp 25 protonated at the inner oxygen, and a direct nucleophilic attack by Asp 25. Strong hydrogen bonds leading to spontaneous proton transfers were observed in both reaction paths. A single-well hydrogen bond was formed between the peptide nitrogen and outer oxygen of Asp 125. The proton was diffusely distributed with an average central position and transferred back and forth on a picosecond scale. In both mechanisms, this interaction helped change the peptide-bond hybridization, increased the partial charge on peptidyl carbon, and in the GA/GB mechanism, helped deprotonate the water molecule. The inner oxygens of the aspartic dyad formed a low-barrier, but asymmetric hydrogen bond; the proton was not positioned midway and made a slightly elongated covalent bond, transferring from one to the other aspartate. In the GA/GB mechanism both aspartates may help deprotonate the water molecule. We observed the breakage of the peptide bond and found that the protonation of the peptidyl amine group was essential for the peptide-bond cleavage. In studies of the direct nucleophilic mechanism, the peptide carbon of the substrate and oxygen of Asp 25 approached as close as 2.3 A. PMID- 14739333 TI - Crystal structure of a tetrameric GDP-D-mannose 4,6-dehydratase from a bacterial GDP-D-rhamnose biosynthetic pathway. AB - d-Rhamnose is a rare 6-deoxy monosaccharide primarily found in the lipopolysaccharide of pathogenic bacteria, where it is involved in host-bacterium interactions and the establishment of infection. The biosynthesis of d-rhamnose proceeds through the conversion of GDP-d-mannose by GDP-d-mannose 4,6-dehydratase (GMD) to GDP-4-keto-6-deoxymannose, which is subsequently reduced to GDP-d rhamnose by a reductase. We have determined the crystal structure of GMD from Pseudomonas aeruginosa in complex with NADPH and GDP. GMD belongs to the NDP sugar modifying subfamily of the short-chain dehydrogenase/reductase (SDR) enzymes, all of which exhibit bidomain structures and a conserved catalytic triad (Tyr-XXX-Lys and Ser/Thr). Although most members of this enzyme subfamily display homodimeric structures, this bacterial GMD forms a tetramer in the same fashion as the plant MUR1 from Arabidopsis thaliana. The cofactor binding sites are adjoined across the tetramer interface, which brings the adenosyl phosphate moieties of the adjacent NADPH molecules to within 7 A of each other. A short peptide segment (Arg35-Arg43) stretches into the neighboring monomer, making not only protein-protein interactions but also hydrogen bonding interactions with the neighboring cofactor. The interface hydrogen bonds made by the Arg35-Arg43 segment are generally conserved in GMD and MUR1, and the interacting residues are highly conserved among the sequences of bacterial and eukaryotic GMDs. Outside of the Arg35-Arg43 segment, residues involved in tetrameric contacts are also quite conserved across different species. These observations suggest that a tetramer is the preferred, and perhaps functionally relevant, oligomeric state for most bacterial and eukaryotic GMDs. PMID- 14739334 TI - The interface of a membrane-spanning leucine zipper mapped by asparagine-scanning mutagenesis. AB - An oligo-leucine sequence has previously been shown to function as an artificial transmembrane segment that efficiently self-assembles in membranes and in detergent solution. Here, a novel technique, asparagine-scanning mutagenesis, was applied to probe the interface of the self-assembled oligo-leucine domain. This novel approach identifies interfacial residues whose exchange to asparagine leads to enhanced self-interaction of transmembrane helices by interhelical hydrogen bond formation. As analyzed by the ToxR system in membranes, the interface formed by the oligo-leucine domain is based on a leucine-zipper-like heptad repeat pattern of amino acids. In general, the strongest impacts on self-assembly were seen with asparagines located around the center of the sequence, indicating that interaction is be more efficient here than at the termini of the transmembrane domains. PMID- 14739335 TI - Breakdown of Bell's theorem for certain objective local parameter spaces. AB - We show that the known proofs of Bell's inequalities contain algebraic manipulations that are not appropriate within the syntax of Kolmogorov's axioms for probability theory without detailed justification. Such justification can be achieved by a variant of the techniques used in Bell-type proofs but only for a subclass of objective local parameter spaces. It cannot be achieved for an extended parameter space that is still objective local and that includes instrument parameters correlated by both time and setting dependencies. PMID- 14739336 TI - Normalization and analysis of cDNA microarrays using within-array replications applied to neuroblastoma cell response to a cytokine. AB - The quantitative comparison of two or more microarrays can reveal, for example, the distinct patterns of gene expression that define different cellular phenotypes or the genes that are induced in the cellular response to certain stimulations. Normalization of the measured intensities is a prerequisite of such comparisons. However, a fundamental problem in cDNA microarray analysis is the lack of a common standard to compare the expression levels of different samples. Several normalization protocols have been proposed to overcome the variabilities inherent in this technology. We have developed a normalization procedure based on within-array replications via a semilinear in-slide model, which adjusts objectively experimental variations without making critical biological assumptions. The significant analysis of gene expressions is based on a weighted t statistic, which accounts for the heteroscedasticity of the observed log ratios of expressions, and a balanced sign permutation test. We illustrated the use of the techniques in a comparison of the expression profiles of neuroblastoma cells that were stimulated with a growth factor, macrophage migration inhibitory factor (MIF). The analysis of expression changes at mRNA levels showed that approximately 99 genes were up-regulated and 24 were reduced significantly (P <0.001) in MIF-stimulated neuroblastoma cells. The regulated genes included several oncogenes, growth-related genes, tumor metastatic genes, and immuno related genes. The findings provide clues as to the molecular mechanisms of MIF mediated tumor progression and supply therapeutic targets for neuroblastoma treatment. PMID- 14739337 TI - Analysis of Pseudomonas aeruginosa 4-hydroxy-2-alkylquinolines (HAQs) reveals a role for 4-hydroxy-2-heptylquinoline in cell-to-cell communication. AB - Bacterial communities use "quorum sensing" (QS) to coordinate their population behavior through the action of extracellular signal molecules, such as the N-acyl l-homoserine lactones (AHLs). The versatile and ubiquitous opportunistic pathogen Pseudomonas aeruginosa is a well-studied model for AHL-mediated QS. This species also produces an intercellular signal distinct from AHLs, 3,4-dihydroxy-2 heptylquinoline (PQS), which belongs to a family of poorly characterized 4 hydroxy-2-alkylquinolines (HAQs) previously identified for their antimicrobial activity. Here we use liquid chromatography (LC)/MS, genetics, and whole-genome expression to investigate the structure, biosynthesis, regulation, and activity of HAQs. We show that the pqsA-E operon encodes enzymes that catalyze the biosynthesis of five distinct classes of HAQs, and establish the sequence of synthesis of these compounds, which include potent cytochrome inhibitors and antibiotics active against human commensal and pathogenic bacteria. We find that anthranilic acid, the product of the PhnAB synthase, is the primary precursor of HAQs and that the HAQ congener 4-hydroxy-2-heptylquinoline (HHQ) is the direct precursor of the PQS signaling molecule. Significantly, whereas phnAB and pqsA-E are positively regulated by the virulence-associated transcription factor MvfR, which is also required for the expression of several QS-regulated genes, the conversion of HHQ to PQS is instead controlled by LasR. Finally, our results reveal that HHQ is itself both released from, and taken up by, bacterial cells where it is converted into PQS, suggesting that it functions as a messenger molecule in a cell-to-cell communication pathway. HAQ signaling represents a potential target for the pharmacological intervention of P. aeruginosa-mediated infections. PMID- 14739338 TI - Genome-wide analysis of gene expression reveals function of the bZIP transcription factor HY5 in the UV-B response of Arabidopsis. AB - The light environment is a key factor that governs a multitude of developmental processes during the entire life cycle of plants. An important and increasing part of the incident sunlight encompasses a segment of the UV-B region (280-320 nm) that is not entirely absorbed by the ozone layer in the stratosphere of the earth. This portion of the solar radiation, which inevitably reaches the sessile plants, can act both as an environmental stress factor and an informational signal. To identify Arabidopsis genes involved in the UV response, we monitored the gene expression profile of UV-B-irradiated seedlings by using high-density oligonucleotide microarrays comprising almost the full Arabidopsis genome (>24,000 genes). A robust set of early low-level UV-B-responsive genes, 100 activated and 7 repressed, was identified. In all cases analyzed, UV-B induction was found to be independent of known photoreceptors. This group of genes is suggested to represent the molecular readout of the signaling cascade triggered by the elusive UV-B photoreceptor(s). Moreover, our analysis identified interactions between cellular responses to different UV-B ranges that led us to postulate the presence of partially distinct but interacting UV-B perception and signaling mechanisms. Finally, we demonstrate that the bZIP transcription factor HY5 is required for UV-B-mediated regulation of a subset of genes. PMID- 14739339 TI - Herpes simplex virus 1 interaction with Toll-like receptor 2 contributes to lethal encephalitis. AB - Human neonates infected with herpes simplex virus 1 (HSV-1) develop one of three distinct patterns of infection: (i) infection limited to the skin, eye or mouth; (ii) infection of the CNS; or (iii) disseminated infection. The disseminated form usually involves the liver, adrenal gland, and lung, and resembles the clinical picture of bacterial sepsis. This spectrum of symptoms in HSV-1-infected neonates suggests that inflammatory cytokines play a significant role in the pathogenesis of the disease. Recent studies suggest that the Toll-like receptors (TLRs) may play an important role in the induction of inflammatory cytokines in response to viruses. TLRs are mammalian homologues of Toll, a Drosophila protein that is essential for host defense against infection. Engagement of TLRs by bacterial, viral, or fungal components leads to the production and release of cytokines and other antimicrobial products. Here, we demonstrate that TLR2 mediates the inflammatory cytokine response to HSV-1 by using both transfected cell lines and knockout mice. Studies of infected mice revealed that HSV-1 induced a blunted cytokine response in TLR2(-/-) mice. Brain levels of monocyte chemoattractant protein 1 chemokine were significantly lower in TLR2(-/-) mice than in either wild-type or TLR4(-/-) mice. TLR2(-/-) mice had reduced mortality compared with wild-type mice. The differences between TLR2(-/-) mice and both wild-type and TLR4(-/-) mice in the induction of monocyte chemoattractant protein 1, brain inflammation, or mortality could not be accounted for on the basis of virus levels. Thus, these studies suggest the TLR2-mediated cytokine response to HSV-1 is detrimental to the host. PMID- 14739340 TI - Cooperation of the ErbB2 receptor and transforming growth factor beta in induction of migration and invasion in mammary epithelial cells. AB - MCF10A mammary epithelial cells form growth-arrested structures when cultured in three-dimensional basement membrane gels. Activation of the receptor tyrosine kinase ErbB2 induces formation of proliferative structures that share properties with noninvasive early stage lesions. We conducted a genetic screen to identify cDNAs that can cooperate with ErbB2 to induce migration in these cells, with the hypothesis that they would represent candidate "second hits" in the development of invasive breast carcinomas. We found that expression of transforming growth factor (TGF)beta1 and TGFbeta3 in cells expressing activated ErbB2 induces migration in transwell chambers and invasive behavior in both basement membrane cultures and invasion chambers. The ability of ErbB2 to cooperate with TGFbeta correlated with sustained, elevated activation of extracellular signal-regulated kinase (Erk)-mitogen-activated protein kinase. Pharmacological reduction of Erk activity inhibited the cooperative effect of TGFbeta and ErbB2 on migration and expression of activated Erk kinase was sufficient to cooperate with TGFbeta to induce migration and invasion, suggesting that sustained Erk activation is critical for ErbB2/TGFbeta cooperation. In addition, we show that costimulation of ErbB2 and TGFbeta induces autocrine secretion of factors that are sufficient to induce migration, but not invasion, by means of both epidermal growth factor receptor-dependent and -independent processes. These results support the role of TGFbeta as a pro-invasion factor in the progression of breast cancers with activated ErbB2 and suggest that activation of the Erk and epidermal growth factor receptor pathways are key in mediating these events. PMID- 14739341 TI - Differentiation and developmental pathways of uropathogenic Escherichia coli in urinary tract pathogenesis. AB - Uropathogenic Escherichia coli (UPEC) are capable of forming complex intracellular bacterial communities (IBC) within the superficial umbrella cells of the bladders of C3H and BALB/c mice. By using time-lapse fluorescence videomicroscopy to observe infected mouse bladder explants, we discovered that IBCs formed by uropathogenic E. coli progressed through four distinct developmental stages that differed with respect to growth rate, bacterial length, colony organization, motility, and its eventual dispersal. In the first phase, bacteria in the IBC were nonmotile, rod shaped, and grew rapidly in loosely organized colonies free in the cytoplasm of the bladder superficial umbrella cells. In the second phase, the loose collection of bacteria in the IBC matured into a slower growing, highly organized biofilm-like community consisting of coccoid bacteria that ultimately filled most of the cytoplasm. In the third phase, bacteria in the biofilm-like state in the IBC switched to a motile rod shaped phenotype allowing detachment from the community and eventual fluxing out of the host cell. During the fourth phase, the bacteria filamented. Filamentation appeared to be in response to a Toll-like receptor 4-mediated innate defense mechanism. Bacteria that fluxed out of the superficial umbrella cells were able to reenter the IBC developmental cascade but with slower kinetics and ultimately a quiescent reservoir was established. Intracellular growth and filamentation provided an advantage to the bacteria in evading infiltrating polymorphonuclear leukocytes. This work has developed a technique to observe live infected organs and revealed a complex differentiation pathway that facilitates bacterial persistence in the urinary tract. PMID- 14739342 TI - DNA A-tract bending in three dimensions: solving the dA4T4 vs. dT4A4 conundrum. AB - DNA A-tracts have been defined as four or more consecutive A.T base pairs without a TpA step. When inserted in phase with the DNA helical repeat, bending is manifested macroscopically as anomalous migration on polyacrylamide gels, first observed >20 years ago. An unsolved conundrum is why DNA containing in-phase A tract repeats of A(4)T(4) are bent, whereas T(4)A(4) is straight. We have determined the solution structures of the DNA duplexes formed by d(GCAAAATTTTGC) [A4T4] and d(CGTTTTAAAACG) [T4A4] with NH(4)(+) counterions by using NMR spectroscopy, including refinement with residual dipolar couplings. Analysis of the structures shows that the ApT step has a large negative roll, resulting in a local bend toward the minor groove, whereas the TpA step has a positive roll and locally bends toward the major groove. For A4T4, this bend is nearly in phase with bends at the two A-tract junctions, resulting in an overall bend toward the minor groove of the A-tract, whereas for T4A4, the bends oppose each other, resulting in a relatively straight helix. NMR-based structural modeling of d(CAAAATTTTG)(15) and d(GTTTTAAAAC)(15) reveals that the former forms a left handed superhelix with a diameter of approximately 110 A and pitch of 80 A, similar to DNA in the nucleosome, whereas the latter has a gentle writhe with a pitch of >250 A and diameter of approximately 50 A. Results of gel electrophoretic mobility studies are consistent with the higher-order structure of the DNA and furthermore depend on the nature of the monovalent cation present in the running buffer. PMID- 14739343 TI - Identification and functional analysis of a locus for improvement of lodging resistance in rice. AB - We identified a new locus responsible for increased pushing resistance of the lower part of rice (Oryza sativa) and analyzed its physiological function to understand how to improve lodging resistance in rice. Quantitative trait loci (QTLs) controlling pushing resistance of the lower part were analyzed in a population of backcross inbred lines of japonica Nipponbare x indica Kasalath plants cut out at 40 cm to exclude the effect of the weight of the upper parts. Five QTLs for pushing resistance were detected; only one QTL from Kasalath on chromosome 5 (prl5) had a positive effect. The likelihood odds ratio curve of prl5 echoed that for lodging resistance by typhoon. We selected three near isogenic lines (NILs) in which the chromosomal region of prl5 was substituted with that of Kasalath in the Nipponbare background. The dry weights and densities and the contents of accumulated carbohydrate in stems below 40 cm (lower stems) in each NIL were significantly higher than those of Nipponbare. There was no difference between Nipponbare and the NILs in yield, root characteristics, or the weights of the upper parts. Pushing resistance of the lower part and lodging resistance in the NILs were up to twice as high as in Nipponbare. These results suggest that prl5 might affect the characteristics of the lower stems of the NILs, thus increasing lodging resistance. PMID- 14739344 TI - Osmotically induced cell swelling versus cell shrinking elicits specific changes in phospholipid signals in tobacco pollen tubes. AB - Pollen tube cell volume changes rapidly in response to perturbation of the extracellular osmotic potential. This report shows that specific phospholipid signals are differentially stimulated or attenuated during osmotic perturbations. Hypo-osmotic stress induces rapid increases in phosphatidic acid (PA). This response occurs starting at the addition of 25% (v/v) water to the pollen tube cultures and peaks at 100% (v/v) water. Increased levels of PA were detected within 30 s and reached maximum by 15 to 30 min after treatment. The pollen tube apical region undergoes a 46% increase in cell volume after addition of 100% water (v/v), and there is an average 7-fold increase in PA. This PA increase appears to be generated by phospholipase D because concurrent transphosphatidylation of n-butanol results in an average 8-fold increase in phosphatidylbutanol. Hypo-osmotic stress also induces an average 2-fold decrease in phosphatidylinositol phosphate; however, there are no detectable changes in the levels of phosphatidylinositol bisphosphates. In contrast, salt-induced hyperosmotic stress from 50 to 400 mm NaCl inhibits phospholipase D activity, reduces the levels of PA, and induces increases in the levels of phosphatidylinositol bisphosphate isomers. The pollen tube apical region undergoes a 41% decrease in cell volume at 400 mm NaCl, and there is an average 2 fold increase in phosphatidylinositol 3,5-bisphosphate and 1.4-fold increase in phosphatidylinositol 4,5-bisphosphate. The phosphatidylinositol 3,5-bisphosphate increase is detected within 30 s and reaches maximum by 15 to 30 min after treatment. In summary, these results demonstrate that hypo-osmotic versus hyperosmotic perturbation and the resultant cell swelling or shrinking differentially activate specific phospholipid signaling pathways in tobacco (Nicotiana tabacum) pollen tubes. PMID- 14739345 TI - Iron-regulated expression of a cytosolic ascorbate peroxidase encoded by the APX1 gene in Arabidopsis seedlings. AB - Iron (Fe) is an essential element for living organisms. However, under aerobic conditions, its use is complicated because of its high insolubility and its potential toxicity through reactivity with reduced forms of oxygen. In plants, Fe overload can lead to intracellular concentrations beyond the storage and detoxification capacities of cells. Such a displacement toward a pro-oxidant state can activate antioxidant defenses, including Fe-mediated expression of ascorbate peroxidase genes. In this work, we demonstrate that Fe overload specifically induces the AtAPX1 gene encoding a cytosolic ascorbate peroxidase in Arabidopsis leaves. The strong constitutive expression of the AtAPX1 gene in roots is unaffected by Fe and depends on the first 5'-untranslated region intron. Presence of an AtAPX1 expressed sequence tag in the Arabidopsis database, longer in its 5' region than what could be predicted from the published AtAPX1transcription initiation site, leads to define a new transcription initiation region for this gene. A minimal promoter sequence enabling Fe-induced expression of the AtAPX1 gene is defined by following expression of various AtAPX1::beta-glucuronidase constructs in transformed Arabidopsis plantlets. This 118-bp minimal promoter sequence contains an Fe-dependent regulatory sequence like cis-element known to be necessary for maize (Zea mays) and Arabidopsis ferritin gene derepression in response to Fe overload. Site-directed mutagenesis of this element within the AtAPX1 promoter sequence does not abolish the Fe dependent activation of a reporter gene, indicating that it is likely not involved in the Fe-regulated expression of the AtAPX1 gene. PMID- 14739346 TI - Nitric oxide functions as a signal in salt resistance in the calluses from two ecotypes of reed. AB - Calluses from two ecotypes of reed (Phragmites communis Trin.) plant (dune reed [DR] and swamp reed [SR]), which show different sensitivity to salinity, were used to study plant adaptations to salt stress. Under 200 mm NaCl treatment, the sodium (Na) percentage decreased, but the calcium percentage and the potassium (K) to Na ratio increased in the DR callus, whereas an opposite changing pattern was observed in the SR callus. Application of sodium nitroprusside (SNP), as a nitric oxide (NO) donor, revealed that NO affected element ratios in both DR and SR calluses in a concentration-dependent manner. N(omega)-nitro-l-arginine (an NO synthase inhibitor) and 2-phenyl-4,4,5,5-tetramethyl-imidazoline-1-oxyl-3-oxyde (a specific NO scavenger) counteracted NO effect by increasing the Na percentage, decreasing the calcium percentage and the K to Na ratio. The increased activity of plasma membrane (PM) H(+)-ATPase caused by NaCl treatment in the DR callus was reversed by treatment with N(omega)-nitro-l-arginine and 2-phenyl-4,4,5,5 tetramethyl-imidazoline-1-oxyl-3-oxyde. Western-blot analysis demonstrated that NO stimulated the expression of PM H(+)-ATPase in both DR and SR calluses. These results indicate that NO serves as a signal in inducing salt resistance by increasing the K to Na ratio, which is dependent on the increased PM H(+)-ATPase activity. PMID- 14739347 TI - High pigment1 mutation negatively regulates phototropic signal transduction in tomato seedlings. AB - Phototropins and phytochromes are the major photosensory receptors in plants and they regulate distinct photomorphogenic responses. The molecular mechanisms underlying functional interactions of phototropins and phytochromes remain largely unclear. We show that the tomato (Lycopersicon esculentum) phytochrome A deficient mutant fri lacks phototropic curvature to low fluence blue light, indicating requirement for phytochrome A for expression of phototropic response. The hp1 mutant that exhibits hypersensitive responses to blue light and red light reverses the impairment of second-positive phototropic response in tomato in phytochrome A-deficient background. Physiological analyses indicate that HP1 functions as a negative regulator of phototropic signal transduction pathway, which is removed via action of phytochrome A. The loss of HP1 gene product in frihp1 double mutant allows the unhindered operation of phototropic signal transduction chain, obviating the need for the phytochrome action. Our results also indicate that the role of phytochrome in regulating phototropism is restricted to low fluence blue light only, and at high fluence blue light, the phytochrome A-deficient fri mutant shows the normal phototropic response. PMID- 14739348 TI - Accumulation of carotenoids and expression of carotenoid biosynthetic genes during maturation in citrus fruit. AB - The relationship between carotenoid accumulation and the expression of carotenoid biosynthetic genes during fruit maturation was investigated in three citrus varieties, Satsuma mandarin (Citrus unshiu Marc.), Valencia orange (Citrus sinensis Osbeck), and Lisbon lemon (Citrus limon Burm.f.). We cloned the cDNAs for phytoene synthase (CitPSY), phytoene desaturase (CitPDS), zeta-carotene (car) desaturase (CitZDS), carotenoid isomerase (CitCRTISO), lycopene beta-cyclase (CitLCYb), beta-ring hydroxylase (CitHYb), zeaxanthin (zea) epoxidase (CitZEP), and lycopene epsilon-cyclase (CitLCYe) from Satsuma mandarin, which shared high identities in nucleotide sequences with Valencia orange, Lisbon lemon, and other plant species. With the transition of peel color from green to orange, the change from beta,epsilon-carotenoid (alpha-car and lutein) accumulation to beta,beta carotenoid (beta-car, beta-cryptoxanthin, zea, and violaxanthin) accumulation was observed in the flavedos of Satsuma mandarin and Valencia orange, accompanying the disappearance of CitLCYe transcripts and the increase in CitLCYb transcripts. Even in green fruit, high levels of beta,epsilon-carotenoids and CitLCYe transcripts were not observed in the juice sacs. As fruit maturation progressed in Satsuma mandarin and Valencia orange, a simultaneous increase in the expression of genes (CitPSY, CitPDS, CitZDS, CitLCYb, CitHYb, and CitZEP) led to massive beta,beta-xanthophyll (beta-cryptoxanthin, zea, and violaxanthin) accumulation in both the flavedo and juice sacs. The gene expression of CitCRTISO was kept low or decreased in the flavedo during massive beta,beta-xanthophyll accumulation. In the flavedo of Lisbon lemon and Satsuma mandarin, massive accumulation of phytoene was observed with a decrease in the transcript level for CitPDS. Thus, the carotenoid accumulation during citrus fruit maturation was highly regulated by the coordination of the expression among carotenoid biosynthetic genes. In this paper, the mechanism leading to diversity in beta,beta-xanthophyll compositions between Satsuma mandarin and Valencia orange was also discussed on the basis of the substrate specificity of beta-ring hydroxylase and the balance of expression between upstream synthesis genes (CitPSY, CitPDS, CitZDS, and CitLCYb) and downstream synthesis genes (CitHYb and CitZEP). PMID- 14739349 TI - Plant and bacterial symbiotic mutants define three transcriptionally distinct stages in the development of the Medicago truncatula/Sinorhizobium meliloti symbiosis. AB - In the Medicago truncatula/Sinorhizobium meliloti symbiosis, the plant undergoes a series of developmental changes simultaneously, creating a root nodule and allowing bacterial entry and differentiation. Our studies of plant genes reveal novel transcriptional regulation during the establishment of the symbiosis and identify molecular markers that distinguish classes of plant and bacterial symbiotic mutants. We have identified three symbiotically regulated plant genes encoding a beta,1-3 endoglucanase (MtBGLU1), a lectin (MtLEC4), and a cysteine containing protein (MtN31). MtBGLU1 is down-regulated in the plant 24 h after exposure to the bacterial signal, Nod factor. The non-nodulating plant mutant dmi1 is defective in the ability to down-regulate MtBGLU1. MtLEC4 and MtN31 are induced 1 and 2 weeks after bacterial inoculation, respectively. We examined the regulation of these two genes and three previously identified genes (MtCAM1, ENOD2, and MtLB1) in plant symbiotic mutants and wild-type plants inoculated with bacterial symbiotic mutants. Plant (bit1, rit1, and Mtsym1) and bacterial (exoA and exoH) mutants with defects in the initial stages of invasion are unable to induce MtLEC4, MtN31, MtCAM1, ENOD2, and MtLB1. Bacterial mutants (fixJ and nifD) and a subset of plant mutants (dnf2, dnf3, dnf4, dnf6, and dnf7) defective for nitrogen fixation induce the above genes. The bacA bacterial mutant, which senesces upon deposition into plant cells, and two plant mutants with defects in nitrogen fixation (dnf1 and dnf5) induce MtLEC4 and ENOD2 but not MtN31, MtCAM1, or MtLB1. These data suggest the presence of at least three transcriptionally distinct developmental stages during invasion of M. truncatula by S. meliloti. PMID- 14739350 TI - Adenylate gradients and Ar:O(2) effects on legume nodules: I. Mathematical models. AB - Mathematical models were developed to test the likelihood that large cytosolic adenylate concentration gradients exist across the bacteria-infected cells of legume nodules. Previous studies hypothesized that this may be the case to account for the unusually low adenylate energy charge (AEC; 0.65) measured in the plant fraction of metabolically active nodules (M.M. Kuzma, H. Winter, P. Storer, I. Oresnik, C.A. Atkins, D.B. Layzell [1999] Plant Physiol 119: 399-407). Simulations coupled leghemoglobin-facilitated O(2) diffusion into the infected cell, through bacteroid nitrogenase activity, with the ATP demand for transport and ammonia assimilation in the plant fraction of ureide- and amide-producing nodules. Although large cytosolic adenylate gradients were predicted to exist in both nodule types, amide nodules were predicted to have steeper AEC gradients (0.82-0.52) than ureide nodules (0.82-0.61). The differences were attributed to an additional ATP demand for Asn synthesis in the amide nodule. Simulations for nodules transferred to an Ar:O(2) atmosphere predicted a major reduction in the magnitude of adenylate gradients and an increase in the AEC of the plant fraction. Results were consistent with a number of experimental studies and were used to propose an experimental test of the models. PMID- 14739351 TI - Wounding enhances expression of AtSUC3, a sucrose transporter from Arabidopsis sieve elements and sink tissues. AB - The Arabidopsis AtSUC3 gene encodes a sucrose (Suc) transporter that differs in size and intron number from all other Arabidopsis Suc transport proteins. Each plant species analyzed so far possesses one transporter of this special type, and several functions have been discussed for these proteins, including the catalysis of transmembrane Suc transport, and also Suc sensing and regulation of other Suc transporters. Here, we show that the AtSUC3 protein is localized in the sieve elements of the Arabidopsis phloem and is not colocalized with the companion cell specific AtSUC2 phloem loader. Even stronger AtSUC3 expression is observed in numerous sink cells and tissues, such as guard cells, trichomes, germinating pollen, root tips, the developing seed coat, or stipules. Moreover, AtSUC3 expression is strongly induced upon wounding of Arabidopsis tissue. The physiological role of AtSUC3 in these different cells and tissues is discussed. PMID- 14739352 TI - CSF removal in infantile posthemorrhagic hydrocephalus results in significant improvement in cerebral hemodynamics. AB - Rational intervention in infants with posthemorrhagic hydrocephalus (PHH) would be facilitated greatly by bedside measure of impaired cerebral perfusion, as there is substantial evidence that impaired perfusion and oxidative metabolism contribute to irreversible brain injury in hydrocephalus. Near-infrared spectroscopy (NIRS) measures changes in the cerebral concentration of oxygenated and deoxygenated hemoglobin and oxidized cytochrome oxidase at the bedside of infants continuously and noninvasively. The total hemoglobin and the hemoglobin difference signal are derived from the sum and difference, respectively, of oxygenated and deoxygenated hemoglobin. Changes in total hemoglobin reflect changes in cerebral blood volume; our previous work has shown that changes in hemoglobin difference signal reflect changes in cerebral blood flow. We hypothesized that cerebrospinal fluid (CSF) removal in infants with PHH would result in significant increases in cerebral perfusion, cerebral blood volume, and oxidative metabolism, as measured by NIRS. Continuous NIRS recordings were performed during CSF removal on 16 infants with PHH. There was a statistically significant increase in oxygenated hemoglobin (p < 0.001), total hemoglobin (p = 0.001), and hemoglobin difference signal (p = 0.006), but not oxidized cytochrome oxidase, accompanying CSF removal. There was no significant correlation between either the volume of CSF removed (in milliliters per kilogram body weight) or the opening pressure and the change in any of the measured or calculated NIRS signals. These findings demonstrate the pronounced effect of CSF removal on cerebral perfusion in infants with PHH. NIRS may be a useful technique to detect impending cerebral ischemia in such infants and thereby provide a means to guide the rational management of PHH. PMID- 14739353 TI - Sonographic biometry of the frontal lobe in normal and growth-restricted neonates. AB - Assessing the impact of restricted intrauterine growth on neonatal frontal lobe (FL) dimensions is important. We aimed to create a sonographic nomogram of FL dimensions in neonates at different gestational ages (GA) and evaluate the impact of small head circumference (HC) on FL dimensions. We conducted sonographic biometry of the FL at birth. We included 218 newborn infants born at GA of 24-43 wk: appropriate for GA and normal HC (n = 178), and small for GA and small HC (n = 23). Infants with a 5-min Apgar score <7, severe congenital malformations, or chromosomal abnormalities were excluded. Through a coronal ultrasound scan via the anterior fontanelle at the level where the most lateral point of the left Sylvian fissure was best demonstrated, we drew a triangle connecting the most lateral point of the Sylvian fissure, the corpus callosum, and the subcalvarian point of the interhemispheric fissure. We measured the three sides of the triangle, Sylvian-fontanellar distance, Sylvian-callosal distance, and fontanellar-callosal distance, and calculated the frontal triangular area. All four FL dimensions increased significantly between 24 and 43 wk of gestation in both appropriate for GA-normal HC and small for GA-small HC neonates, and were strongly correlated with HC and birth weight. Regression lines of GA against Sylvian-fontanellar distance, Sylvian-callosal distance, fontanellar-callosal distance, and frontal triangular area in the appropriate for GA-normal HC group differed significantly from those of the small for GA-small HC group (p < 0.05). Male neonates had significantly larger Sylvian-fontanellar and Sylvian-callosal distances than females (p < 0.01 and p < 0.015, respectively). In conclusion, FL measures increased significantly between 24 and 43 wk of gestation, and were strongly correlated with HC. We speculate that a sonographically small fetal HC implies growth restriction of the fetal FL. PMID- 14739354 TI - Prolonged neural expiratory time induced by mechanical ventilation in infants. AB - Mechanical ventilation may interfere with the spontaneous breathing pattern in infants because they have strong reflexes that play a large role in the control of breathing. This study aimed to answer the following questions: does a ventilator-assisted breath 1) reduce neural inspiratory time, 2) reduce the amplitude of the diaphragm electrical activity, and 3) prolong neural expiration, within the delivered breath? In 14 infants recovering from acute respiratory failure (mean age and weight were 2.3 +/- 1.3 mo and 3.95 +/- 0.82 kg, respectively), we measured 1) the electrical activity of the diaphragm with a multiple-array esophageal electrode, and 2) airway opening pressure, while patients breathed on synchronized intermittent mandatory ventilation (mean rate, 11.2 +/- 6.5 breaths/min). We compared neural inspiratory and expiratory times for the mandatory breaths and for the spontaneous breaths immediately preceding and following the mandatory breath. Although neural inspiratory time was not different between mandatory and spontaneous breaths, neural expiratory time was significantly increased (p < 0.001) for the mandatory breaths (953 +/- 449 ms) compared with the premandatory and postmandatory spontaneous breaths (607 +/- 268 ms and 560 +/- 227 ms, respectively). Delivery of the mandatory breath resulted in a reduction in neural respiratory frequency by 28.6 +/- 6.4% from the spontaneous premandatory frequency. The magnitude of inspiratory electrical activity of the diaphragm was similar for all three breath conditions. For the mandatory breaths, ventilatory assist persisted for 507 +/- 169 ms after the end of neural inspiratory time. Infant-ventilator asynchrony (both inspiratory and expiratory asynchrony) was present in every mandatory breath and constituted 53.4 +/- 26.2% of the total breath duration. PMID- 14739355 TI - Clinical features of the metabolic syndrome in adolescents: minor role of the Trp64Arg beta3-adrenergic receptor gene variant. AB - Obesity and hypertension are increasing medical problems in adolescents. We evaluated the association between being overweight-particularly abdominal fat-and having hypertension and assessed the contribution of the Trp64Arg beta3 adrenergic receptor gene variant. In a population-based study, we determined family history, anthropometric variables, and arterial blood pressure of 934 high school students, out of whom we selected 121 normotensive and 54 hypertensive students. Biochemical measurements included circulating renin and angiotensin converting enzyme activities, leptin, glucose, insulin and lipid levels, and beta3-adrenergic receptor genotypes. We used Mann-Whitney U test, chi2-test, and Spearman rank-order correlation. In the total population, hypertension prevalence increased across the entire range of body mass index (BMI) percentiles. In the sample, hypertensive students showed higher BMI, waist-to-hip ratio, triglycerides, and insulin resistance and lower HDL-cholesterol than normotensive students did. Age- and sex-adjusted systolic arterial blood pressure was correlated with BMI, waist-to-hip ratio, insulin resistance, and leptin. Leptin was correlated with BMI and homeostasis model assessment method. We found no association among hypertension, BMI, and leptin levels with beta3-adrenergic receptor genotypes. Especially in girls, the waist-to-hip ratio was, however, suggestively higher in Arg64 variant carriers than in noncarriers, independent of hypertension. In fact, there was a significantly (p < 0.01) higher frequency of carriers of the Arg64 variant across the waist-to-hip ratio quartiles. In adolescents of European origin, hypertension is associated with an increased degree of obesity among other characteristics of the metabolic syndrome; the Trp64Arg variant of the beta3-adrenergic receptor gene may favor the central adiposity gain. PMID- 14739356 TI - Abnormal heart rate characteristics are associated with neonatal mortality. AB - Estimating the risk of in-hospital mortality in the newborn intensive care unit can provide important information for health-care providers, and illness severity scores have been devised to provide mortality risk estimates. Calculation of illness severity scores is time-consuming, and the information used to predict mortality is collected only for the first 12 to 24 h of life. A noninvasive continuous measure that uses information collected throughout the hospitalization and that requires no data entry could be less costly and more informative. We have previously shown that the abnormal heart rate characteristics (HRC) of reduced variability and transient decelerations accompany neonatal illness such as late-onset sepsis. We hypothesized that more frequent and severe abnormal HRC are associated with an increased risk of death. We tested this hypothesis in two ways. Using data on infants older than 7 d of age, we first determined the association of the HRC index with death in the next week. Second, we devised a cumulative HRC score and determined its association with in-hospital death. There were 37 deaths in the 685 patients. The major findings were 1) the HRC index showed highly significant association with death in the succeeding 7 d (receiver operating characteristic area > 0.7, p < 0.001), and 2) the cumulative HRC was highly significantly associated with neonatal in-hospital mortality (receiver operating characteristic area > 0.80, p < 0.001). In both analyses, HRC added information to birth weight, gestational age, and postnatal age (p < 0.01). The HRC index provides independent information about the risk of neonatal death in the upcoming 7 d, and the cumulative HRC is an estimate of the risk of in hospital neonatal mortality. PMID- 14739357 TI - Stereospecific regulation of tyrosine hydroxylase and proenkephalin genes by short-chain fatty acids in rat PC12 cells. AB - Circulating short-chain fatty acids (SCFAs) are primarily derived from bacterial fermentation of carbohydrates in the colon where they function as physiologic modulators of epithelial cell maturation. Butyrate has been shown to induce tyrosine hydroxylase, the rate-limiting enzyme of catecholamine synthesis, and enkephalin neuropeptide gene transcription, suggesting a role in perinatal sympathoadrenal stress-adaptation. We sought to determine whether there were SCFA structural requirements for this effect. Nine biologically relevant SCFAs and butyrate derivatives were tested in an in vitro model (PC12, rat pheochromocytoma cells) for their ability to regulate neurotransmitter-related gene expression. Our results revealed that among all the studied SCFAs, only propionate and butyrate increased tyrosine hydroxylase and proenkephalin mRNA levels. The functional activity was selective to the carbon atom chain length and associated with the presence of an ethyl moiety in the carbon atom backbone chain. Modifications or absence of this domain affected the gene induction response, suggesting a receptor-mediated mechanism(s). Moreover, propionate, butyrate, and the drug 4-phenylbutyrate were each shown to regulate transmitter genes via at least three independent mechanisms: histone hyperacetylation, cAMP signaling, or peroxisome proliferator-activated receptor gamma-mediated pathways. Thus, the biologic impact of SCFAs on catecholaminergic and opioid systems depend on the activation of SCFA-specific, dose-specific, and gene-specific molecular mechanisms. We speculate that 1) circulating levels of SCFAs may influence sympathoadrenal transmitter biosynthesis and hence whole animal stress-adaptive responsiveness after birth, and 2) the adverse effects of antibiotics on delayed acquisition of postnatal gut flora may affect this apparent evolutionary advantage of gut colonization. PMID- 14739360 TI - Evidence of exercise-induced arterial hypoxemia in prepubescent trained children. AB - Exercise-induced arterial hypoxemia (EIAH) is a recognized phenomenon in highly trained adults. Like adult athletes, prepubescent trained children may develop high-level metabolic demand but with a limited lung capacity in comparison with adults. The purpose of this investigation was to search for evidence of EIAH in prepubescent trained children. Twenty-four prepubescent (age: 10.3 +/- 0.2 y) trained children (10.0 +/- 0.7 h of weekly physical activity) performed pulmonary function tests and a graded maximal exercise test on a cycle ergometer. EIAH was defined as a drop of at least 4% from resting level arterial oxygen saturation (Sao(2)) measured by pulse oximetry. EIAH was observed in seven children. Forced vital capacity (FVC), ventilatory response to exercise (Delta(E)/Deltaco(2)), and breathing reserve at maximal exercise were significantly lower, whereas tidal volume relative to FVC was higher in hypoxemic children than in nonhypoxemic children; weekly physical activity and maximal oxygen uptake were similar. Moreover, positive relationships were found between Sao(2) at maximal exercise and breathing reserve (r = 0.56; p < 0.05) or volume relative to FVC (r = 0.70; p < 0.01). EIAH may occur in prepubescent trained children with a relatively low maximal oxygen uptake (42 mL. min(-1). kg(-1)); however, the mechanisms remain unclear and need to be investigated more accurately. PMID- 14739359 TI - Genetics and early disturbances of breathing control. AB - Early disturbances in breathing control, including apneas of prematurity and apparently life-threatening events, account for some cases of sudden infant death syndrome and for a rare disorder called congenital central hypoventilation syndrome (CCHS). Data suggesting a genetic basis for CCHS have been obtained. Recently, we found heterozygous de novo mutations of the PHOX2B gene in 18 of 29 individuals with CCHS. Most mutations consisted of five to nine alanine expansions within a 20-residue polyalanine tract, probably resulting from nonhomologous recombination. Other mutations, generally inherited from one of the parents, in the coding regions of genes involved in the endothelin and RET signaling pathways and in the brain-derived-neurotrophic factor (BDNF) gene have been found in a few CCHS patients. Interestingly, all these genes are involved in the development of neural crest cells. Targeted disruption of these genes in mice has provided information on the pathophysiological mechanisms underlying CCHS. Despite the identification of these genes involved in breathing control, none of the genetically engineered mice developed to date replicate the full human CCHS respiratory phenotype. Recent insights into the genetic basis for CCHS may shed light on the genetics of other early disturbances in breathing control, such as apnea of prematurity and sudden infant death syndrome. PMID- 14739361 TI - A short history of pediatric endocrinology in North America. AB - Pediatric endocrinology evolved as a subspecialty from the era of biochemical and metabolic clinical investigation led by John Howland, Edwards Park, and James Gamble at Johns Hopkins; Allan Butler at Boston University and Harvard University; Daniel Darrow at Yale University; and Irving McQuarrie at the University of Rochester and the University of Minnesota during the early 20th century. The father of the new subspecialty was Lawson Wilkins, a private pediatric practitioner in Baltimore, Maryland, who was invited by Dr. Edwards Park to establish an endocrine clinic at the Harriet Lane Home at Johns Hopkins in 1935. Dr. Wilkins managed his practice and the clinic until 1946, when, at the age of 52, he accepted a full-time position at the University. Dr. Nathan Talbot was invited to develop a pediatric endocrine clinic at Massachusetts General Hospital by Allan Butler in 1942. These units and their associated subspecialty training programs during the 1950s and 1960s provided the large majority of the second-generation pediatric endocrinologists who went on to establish endocrine subspecialty programs in university medical centers in North America as well as Europe and South America. Diabetes as a clinical pediatric discipline evolved in parallel from the early clinics of Elliott Joslin and Priscilla White in Boston, M.C. Hardin and Robert Jackson at the University of Iowa, George Guest at the University of Cincinnati Children's Hospital, and Alex Hartman at the St. Louis Children's Hospital. The Lawson Wilkins Pediatric Endocrine Society was founded in 1971, and the Council on Diabetes and Youth was established within the American Diabetes Association in 1980. Medical and economic factors led to increasing integration of pediatric diabetes and general endocrine care and training, and diabetes care now is a major activity within the subspecialty of pediatric endocrinology. The growth of pediatric endocrinology in North America has paralleled the growth of academic medicine during the past half-century. In 2002, there were 72 training programs in North America: 65 in the United States and seven in Canada. The endocrinology sub-board of the American Board of Pediatrics was established in 1978 to certify training and competence in endocrinology, including diabetes. By 2002, the board had certified 927 pediatric endocrinologists. Pediatric endocrine subspecialists during the past half-century have contributed major advances in our understanding of the ontogeny of endocrine systems and the diagnosis and treatment of fetal-perinatal endocrine disorders; newborn screening for endocrine and metabolic disorders; the physiology and therapies for disorders of sexual differentiation and pubertal maturation; the development of anthropometric standards for childhood growth and development; the characterization and physiology of hormone systems, including receptors and hormone actions; the molecular genetics of a number of congenital endocrine disorders and heritable endocrine diseases; development of pediatric endocrine diagnostics and reference standards; the pathophysiology and management of autoimmune endocrine disease; and development of a growing armamentarium of therapeutic agents for treatment of endocrine and metabolic diseases. PMID- 14739362 TI - Different responses of myocardial and cerebral blood flow to cord occlusion in exteriorized fetal sheep. AB - Type and duration of fetal asphyxial insult affect the distribution of blood flow to the heart and brain. The purpose of this study was to describe dynamic and quantitative changes in regional myocardial and cerebral blood flow (CBF) during fetal asphyxia induced by total occlusion of the umbilical cord. Eleven exteriorized fetal sheep were subjected to total umbilical cord occlusion and five fetal sheep served as sham controls. Regional blood flow (BF) to the brain and heart was quantified using radioactive microspheres before and after 5 min of occlusion and finally when fetal mean arterial blood pressure had decreased below 25 mm Hg, 9.8 (0.8) [mean (SD)] min after occlusion. Right coronary arterial (RCA) blood flow velocity and carotid BF were registered continuously. Mean values of arterial pH and oxygen content (mL O(2)/100 mL) were 7.08 (0.11) and 4.4 (2.9) before cord occlusion and decreased to 6.83 (0.05) and 1.4 (0.9) at 5 min after occlusion (p < 0.01, respectively). Carotid BF was significantly below preocclusion values by 2.5 min (p < 0.05), whereas RCA velocity time integral per minute remained above preocclusion values for 9 min. CBF decreased from 316 (24) before cord occlusion to 156 (30) mL/min/100 g at 5 min (p < 0.01), whereas right myocardial BF was maintained at 792 (125) and 751 (183) mL/min/100 g, respectively. CBF decreased rapidly after total cord occlusion whereas myocardial BF increased and was maintained until shortly before cardiac arrest, suggesting the myocardium to be better preserved during this type of insult in already partially asphyxiated fetuses. PMID- 14739363 TI - Neonatal hypoxia triggers transient apoptosis followed by neurogenesis in the rat CA1 hippocampus. AB - Continuous generation of new neurons has been demonstrated in the adult mammalian brain, and this process was shown to be stimulated by various pathologic conditions, including cerebral ischemia. Because brain oxygen deprivation is particularly frequent in neonates and represents the primary event of asphyxia, we analyzed long-term consequences of transient hypoxia in the newborn rat. Within 24 h after birth, animals were exposed to 100% N(2) for 20 min at 36 degrees C, and temporal changes in the vulnerable CA1 hippocampus were monitored. Cell density measurements revealed delayed cell death in the pyramidal cell layer reflecting apoptosis, as shown by characteristic nuclear morphology and expression levels of Bcl-2, Bax, and caspase-3. Neuronal loss was confirmed by reduced density of neuron-specific enolase (NSE)-labeled cells, and peaked by 1 wk post insult, to reach 27% of total cells. A gradual recovery then occurred, and no significant difference in cell density could be detected between controls and hypoxic rats at postnatal d 21. Repeated injections of bromodeoxyuridine (50 mg/kg) showed that newly divided cells expressing neuronal markers increased by 225% in the germinative subventricular zone, and they tended to migrate along the posterior periventricle toward the hippocampus. Therefore, transient hypoxia in the newborn rat triggered apoptosis in the CA1 hippocampus followed by increased neurogenesis and apparent anatomical recovery, suggesting that the developing brain may have a high capacity for self-repair. PMID- 14739364 TI - Test-retest reliability of swept visual evoked potential measurements of infant visual acuity and contrast sensitivity. AB - The aim of the study was to describe variations in swept visual evoked potential (SWEEP-VEP) assessment of visual acuity and contrast sensitivity in infants and to evaluate the best way to estimate visual performance from obtained SWEEP-VEP data. The visual performance of 92 infants (6-40 wk of age) was measured in two separate visits. Results were verified with repeated tests in seven adults. There was a strong association between the two measurements of infant visual acuity (r = 0.91, p < 0.001), with no constant bias and an inter-assay coefficient of variation of 8.4%. The intra-assay coefficient of variation was 17% and in repeated sessions all obtained acuity measures were normally distributed, indicating that the mean and not the maximum threshold best estimates visual acuity. This estimate of visual acuity also had lower test-retest variability than those calculated from the maximum threshold or threshold from the average EEG signals (p = 0.001). Test-retest measures of infant contrast sensitivity had a correlation coefficient of 0.72 (p < 0.001) and an inter-assay coefficient of variation of 23%. With the observed test-retest variability, SWEEP-VEP is less valid for estimating the visual performance of individual subjects, but it can give reliable group means. This method was well suited to describe visual development in the infants, which for acuity as well as contrast sensitivity increased by 0.64 octave per doubling in age. However, the variability of the SWEEP-VEP method can be a limiting factor, for example, in the assessment of the potential effect of dietary docosahexaenoic acid in a homogeneous group of infants. PMID- 14739365 TI - Timing of neutrophil depletion influences long-term neuroprotection in neonatal rat hypoxic-ischemic brain injury. AB - In neonatal rats, neutrophils do not accumulate in ischemic brain parenchyma to the extent that they do in adult rodents. They are also confined to the intravascular compartment during the first few hours of recovery. However, neonatal rats rendered neutropenic have less brain swelling after a hypoxic ischemic (HI) insult. In this study, we used the Rice-Vannucci model of HI brain injury in 7-d-old rats, and we depleted neutrophils before injury in one group and 4-8 h after injury in another group to determine 1) whether neutrophils contribute to cerebral atrophy, 2) whether neutropenia induced within 8 h after recovery from HI is neuroprotective, and 3) whether neutropenia preserved energy metabolites during the HI insult. Brain energy metabolites were measured at 0 h and 6 h of recovery. Brain atrophy was measured morphometrically on brain slices at 2 wk of recovery. In 67 rats, we found that neutropenia induced before the HI insult, but not after HI, reduced brain swelling at 42 h of recovery by about 75% (p < 0.001). In another 60 rats, we found that cerebral atrophy was reduced by 61% provided that neutropenia was induced before HI (p < 0.05). Total adenine nucleotides were better preserved in the neutropenic rats at the end of the HI insult (0 h recovery); p < 0.05. We conclude that neutrophils do contribute to vascular dysfunction either during the HI insult or early hours (<4-8 h) of recovery. Antineutrophil strategies initiated after this time are unlikely to be protective in the neonatal rat. PMID- 14739366 TI - Tie2-Cre-induced inactivation of a conditional mutant Nf1 allele in mouse results in a myeloproliferative disorder that models juvenile myelomonocytic leukemia. AB - Neurofibromatosis type one (NF1) is a common genetic disorder affecting 1:4000 births and is characterized by benign and malignant tumors. Children with NF1 are predisposed to juvenile myelomonocytic leukemia. The Nf1 gene encodes neurofibromin, which can function as a Ras GTPase-activating protein. Neurofibromin deficiency in mice leads to mid-gestation lethality due to cardiovascular defects. We have previously shown that conditional inactivation of Nf1 using Tie2-Cre recapitulates the heart defects seen in Nf1(-/-) embryos. Tie2 Cre transgenic mice express Cre recombinase in all endothelial cells. Here, we show that Tie2-Cre-mediated deletion of Nf1 also leads to excision of Nf1 in the hematopoietic lineage. Surviving mice exhibit a myeloproliferative disorder similar to juvenile myelomonocytic leukemia seen in NF1 patients. These mice provide a useful model to study neurofibromin deficiency in hematopoiesis. Furthermore, defects in Tie2-Cre-expressing progenitors that result in heart and blood defects suggest that related heart and blood disorders in NF1 and other syndromes represent disorders of the hemangioblast. PMID- 14739367 TI - Genetic screening for susceptibility to infection in the NICU setting. PMID- 14739368 TI - Lung inflammatory responses to intratracheal interleukin-1alpha in ventilated preterm lambs. AB - Interleukin-1alpha is an early response proinflammatory cytokine that has been associated with chorioamnionitis and preterm labor, brain injury, and bronchopulmonary dysplasia. However, IL-1alpha also can increase expression of surfactant proteins and induce lung maturation in the preterm fetus. We measured the effects of IL-1alpha given by intratracheal instillation (IT) and compared the responses with injection of i.v. IL-1alpha in surfactant-treated and ventilated premature lambs. IT recombinant ovine IL-1alpha at doses of 5 and 50 microg/kg caused a similar large recruitment of neutrophils into the bronchoalveolar lavage fluid. The neutrophils expressed CD11b, CD14, and CD44, but did not produce increased amounts of H(2)O(2). Cells from the bronchoalveolar lavage fluid had increased expression of proinflammatory cytokines, which also were increased in mRNA from lung tissue. The IT IL-1alpha also suppressed the expression of surfactant protein-C mRNA. Systemic effects were decreased neutrophils in blood, decreased lung function, increased heart rate, and hypotension or death in the 50 microg/kg IL-1alpha IT group and only decreased neutrophils in the blood in the 5 microg/kg IL-1alpha IT group. The i.v. IL 1alpha caused no lung inflammation or injury but did result in severe neutropenia and hypotension leading to early death. IT IL-1alpha can cause intense lung inflammation and systemic shock in ventilated preterm lungs. PMID- 14739369 TI - Prenatal exposure to IL-1beta results in disturbed skeletal growth in adult rat offspring. AB - Events occurring early in life or prenatally are able to play important roles in the pathogenesis of diseases in adult life. Different sorts of stress or hormonal influences, during particular periods of pregnancy, may result in persistent or transient changes in physiology. IL-1 is a multifunctional cytokine that is involved in bone metabolism. The aim of the present study was to investigate whether exposure to IL-1beta during fetal life has any effect on skeletal growth or bone mineral density in adult rat offspring. Pregnant rats were given intraperitoneal injections of IL-1beta, 1 microg/rat, or saline on days 8, 10, and 12 of gestation. Male IL-1-exposed offspring showed reduced height, areal bone mineral density, and bone mineral content at vertebra L5. Tibial length was reduced in both male and female offspring. Peripheral quantitative computed tomography analyses revealed reduced cortical bone mineral content caused by a decreased cortical cross-sectional area as a result of a decreased cortical thickness, whereas there was no reduction in the amount of trabecular bone in the tibia of male offspring. Our results demonstrate that prenatal exposure to IL-1 can induce specific programming of skeletal tissue. In conclusion, prenatal IL-1 exposure results in decreased skeletal growth and a reduced amount of cortical bone but unchanged trabecular bone mineral density in adult rat offspring. PMID- 14739370 TI - Mutations of genes involved in the innate immune system as predictors of sepsis in very low birth weight infants. AB - Mutations of genes involved in the innate immune system have been reported to be associated with an increased sepsis rate in adults. We determined the -159T mutation of the CD14 gene, the 896G mutation of the toll-like receptor 4 gene, the 3020insC mutation of the NOD2 gene (NOD2-3020insC), the IL-6 174G/C promoter polymorphism (IL6-174G/C), and the mannose-binding lectin genotype and their association to the subsequent development of neonatal sepsis in a large cohort of very low birth weight (VLBW) infants. Fifty (14%) of 356 VLBW infants developed blood culture-proven sepsis during their stay in the hospital. VLBW infants carrying the NOD2-3020insC allele (n =15) and the IL6-174G allele (n =121) had a significantly higher rate of blood culture-proven sepsis (33% and 19.8%, respectively) than VLBW infants without these genotypes (p = 0.046 and 0.035, respectively). In a multivariate logistic regression analysis, gestational age less than 28 wk (odds ratio, 3.2; 95% confidence interval, 1.7-6.0; p < 0.001) and the homozygous IL6-174G allele (odds ratio, 1.9; 95% confidence interval, 1.0 3.9; p = 0.039) were predictive for the development of sepsis, whereas the NOD2 3020insC allele was only of borderline significance (odds ratio, 3.2; 95% confidence interval, 1.0-10.4; p = 0.052). VLBW infants with repeated episodes of sepsis had higher frequencies of the NOD2-3020insC and IL6-174G allele. The increased sepsis rate of homozygous IL6-174G carriers was especially related to an increase in Gram-positive infections, and was not observed in VLBW infants who received prophylaxis with teicoplanin (frequency of Gram-positive sepsis in homozygous IL6-174G carriers without prophylaxis 16.5% versus 2.4% in homozygous IL6-174G carriers with prophylaxis; p = 0.033). PMID- 14739371 TI - Does in utero exposure to heavy maternal smoking induce nicotine withdrawal symptoms in neonates? AB - Maternal drug use during pregnancy is associated with fetal passive addiction and neonatal withdrawal syndrome. Cigarette smoking-highly prevalent during pregnancy is associated with addiction and withdrawal syndrome in adults. We conducted a prospective, two-group parallel study on 17 consecutive newborns of heavy-smoking mothers and 16 newborns of nonsmoking, unexposed mothers (controls). Neurologic examinations were repeated at days 1, 2, and 5. Finnegan withdrawal score was assessed every 3 h during their first 4 d. Newborns of smoking mothers had significant levels of cotinine in the cord blood (85.8 +/- 3.4 ng/mL), whereas none of the controls had detectable levels. Similar findings were observed with urinary cotinine concentrations in the newborns (483.1 +/- 2.5 microg/g creatinine versus 43.6 +/- 1.5 microg/g creatinine; p = 0.0001). Neurologic scores were significantly lower in newborns of smokers than in control infants at days 1 (22.3 +/- 2.3 versus 26.5 +/- 1.1; p = 0.0001), 2 (22.4 +/- 3.3 versus 26.3 +/- 1.6; p = 0.0002), and 5 (24.3 +/- 2.1 versus 26.5 +/- 1.5; p = 0.002). Neurologic scores improved significantly from day 1 to 5 in newborns of smokers (p = 0.05), reaching values closer to control infants. Withdrawal scores were higher in newborns of smokers than in control infants at days 1 (4.5 +/- 1.1 versus 3.2 +/- 1.4; p = 0.05), 2 (4.7 +/- 1.7 versus 3.1 +/- 1.1; p = 0.002), and 4 (4.7 +/- 2.1 versus 2.9 +/- 1.4; p = 0.007). Significant correlations were observed between markers of nicotine exposure and neurologic-and withdrawal scores. We conclude that withdrawal symptoms occur in newborns exposed to heavy maternal smoking during pregnancy. PMID- 14739372 TI - Impaired fibrinolytic activity is present in children with dyslipidemias. AB - Dyslipidemias are major risk factors for atherosclerosis and cardiovascular disease. Abnormalities of fibrinolytic and coagulation components are considered useful predictors of cardiovascular morbidity and mortality in adults. This study examined whether fibrinolytic and coagulation components are abnormal in children with dyslipidemia. Thirty-six children with asymptomatic dyslipidemia, and 26 control subjects underwent venous occlusion stress testing with collection of preocclusion and postocclusion blood samples. All samples were assayed for tissue plasminogen activator, plasminogen, plasminogen activator inhibitor-1, alpha(2) antiplasmin, alpha(2)-macroglobulin, D-dimer, fibrinogen, and von Willebrand factor. Children with dyslipidemia had significantly decreased levels of tissue plasminogen activator in both preocclusion and postocclusion samples compared with control subjects, reflecting decreased fibrinolytic activity. Children with dyslipidemia also had significantly increased levels of plasminogen, alpha(2) macroglobulin, and fibrinogen in preocclusion and postocclusion samples compared with control subjects. In conclusion, decreased fibrinolytic activity is present in asymptomatic children with dyslipidemias, potentially reflecting endothelial dysfunction and increased risk of cardiovascular disease in early adult life. Further studies are required to determine the usefulness of this marker in predicting disease progression or response to therapy. PMID- 14739373 TI - One hundred years of anthrax. PMID- 14739375 TI - The dissemination of anthrax from imported wool: Kidderminster 1900-14. AB - BACKGROUND: A century ago anthrax was a continuing health risk in the town of Kidderminster. The distribution of cases in people and in animals provides an indication of the routes by which spores were disseminated. The response to these cases provides an insight into attitudes to an occupational and environmental risk at the time and can be compared with responses in more recent times. AIMS: To assess the distribution of anthrax cases associated with the use of contaminated wool and to review the response to them. METHODS: The area studied was Kidderminster, Worcestershire, England, from 1900 to 1914. Data sources were national records of the Factory Inspectorate and local records from the infirmary, Medical Officer of Health and inquest reports, and county agricultural records, supplemented by contemporary and later review articles. Case reports and summary data were analysed, and discussions and actions taken to improve precautions reviewed. RESULTS: There were 36 cases of anthrax, with five deaths, one of which was the sole case of the internal form of the disease. Cases of cutaneous anthrax were most frequently found in those handling raw wool, but they also occurred in workers at later stages of the spinning process and in people with little or no recorded exposure to contaminated wool. Limited precautionary measures were in place at the start of the study period. Some improvements were made, especially in the treatment of infections, but wool with a high risk of anthrax contamination continued to be used and cases continued to arise. Major changes were made to the disposal of waste and to agricultural practice in contaminated areas to curtail outbreaks in farm animals. CONCLUSIONS: The introduction of anthrax as a contaminant of imported wool led not only to cases in the highly exposed groups of workers but also to cases in other members of the population and in farm animals. The measures taken during the study period reduced fatalities from cutaneous anthrax but did not eliminate the disease. Public concern about the cases was muted. PMID- 14739374 TI - The transmission of tuberculosis in the light of new molecular biological approaches. AB - This review briefly summarises the recent achievements in tuberculosis epidemiology associated with the introduction of molecular methods, and considers the implications of these methods for the understanding of occupational tuberculosis transmission. Special attention is paid to the relative contribution of recently transmitted tuberculosis; risk factors for recent transmission; and the occurrence and frequency of exogenous reinfection. There is a need for occupational epidemiological studies, which should combine the methods of "classical" epidemiology with those of molecular epidemiology. PMID- 14739376 TI - Lung cancer mortality in UK nickel-cadmium battery workers, 1947-2000. AB - AIMS: To investigate mortality from lung cancer in nickel-cadmium battery workers in relation to cumulative exposure to cadmium hydroxide. METHODS: The mortality of a cohort of 926 male workers from a factory engaged in the manufacture of nickel-cadmium batteries in the West Midlands of England was investigated for the period 1947-2000. All subjects were first employed at the plant in the period 1947-75 and employed for a minimum period of 12 months. Work histories were available for the period 1947-86; the factory closed down in 1992. Two analytical approaches were used, indirect standardisation and Poisson regression. RESULTS: Based on serial mortality rates for the general population of England and Wales, significantly increased mortality was shown for cancers of the pharynx (observed (Obs) 4, expected (Exp) 0.7, standardised mortality ratio (SMR) 559, p<0.05), non malignant diseases of the respiratory system (Obs 61, Exp 43.0, SMR 142, p<0.05), and non-malignant diseases of the genitourinary system (Obs 10, Exp 4.1, SMR 243, p<0.05). Non-significantly increased SMRs were shown for lung cancer (Obs 45, Exp 40.7, SMR 111) and cancer of the prostate (Obs 9, Exp 7.5, SMR 116). Estimated cumulative cadmium exposures were not related to risks of lung cancer or risks of chronic obstructive pulmonary diseases, even when exposure histories were lagged first by 10, then by 20 years. CONCLUSIONS: The study findings do not support the hypotheses that cadmium compounds are human lung carcinogens. PMID- 14739377 TI - Cutaneous melanoma: hints from occupational risks by anatomic site in Swedish men. AB - AIMS: To improve knowledge of the epidemiology of melanoma by comparing occupational risks of cutaneous melanoma (CM) by anatomic site in Swedish workers. METHODS: Male workers employed in 1970 and living in the country in 1960 were followed up from 1971 to 1989 using the Swedish Registers of Death and Cancer. A more specifically exposed subcohort included men reporting the same occupation in 1960 and 1970. For each location, occupational risk ratios (RRs) were extracted from Poisson regression models adjusted by age, period, town size, and geographical area. To diminish the influence of socioeconomic factors, intrasector analyses, comparing only jobs belonging to the same occupational sector, were performed. Risk patterns for different locations were compared. RESULTS: High RRs for different sites were found among workers exposed to UV sources (dentists, physiotherapists, and lithographers), and sun exposed workers (harbour masters, and lighthouse/related work). Risk excesses were seen in fur tailors, tanners/fur dressers, patternmakers/cutters, electrical fitters/wiremen, telephone/telegraph installers/repairmen, and some glass/pottery/tile workers. Results for lower and upper limbs were significantly correlated but somewhat independent of those found in thorax, the most frequent location. Correlation between head/neck and thorax was moderate. Specific risk excesses were found for rolling mill workers in head/neck, for chimney sweeps in upper limbs, and for aircraft pilots/navigators/flight engineers in lower limbs. CONCLUSIONS: High RRs in the trunk among occupations with UV exposure from artificial sources suggest an effect not restricted to exposed sites. An unusual distribution of cases and RRs in chimney sweeps, rolling-mill, or glass/pottery/tile workers suggests local effects of exposures. The not previously reported risk excess in this job and in fur related processes, and the RR in electrical fitters and telephone/telegraph installers deserve further investigation. Disparities between locations, as RRs in thorax and limbs, may reflect differences in aetiological mechanisms. PMID- 14739378 TI - A six year follow up study of the subclinical effects of carbon disulphide exposure on the cardiovascular system. AB - AIMS: A six year prospective cohort study was conducted to clarify whether the current carbon disulphide (CS2) exposure level is low enough to prevent subclinical health impairment and/or to ameliorate health effects due to previous high exposure. This paper describes the effects on the cardiovascular systems. METHODS: The study subjects were 432 male workers exposed to CS2 and 402 non exposed workers in Japan, all of whom were examined in 1992-93. A total of 251 CS2 exposed, 140 formerly exposed, and 359 non-exposed workers participated in the follow up survey (follow up rate 89.9%) in 1998-99. Mean duration of exposure was 19.3 years at the end the study. Mean CS2 and 2-thiothiazolidine-4-carboxylic acid (TTCA) concentrations were 5.0 ppm and 1.6 mg/g creatinine. Health items examined were serum biochemical indices including lipids and coagulation fibrinolysis factors, blood pressure, aortic stiffness, ophthalmography, and electrocardiography at rest and after Master's double 2 step test. Potential confounding factors were adjusted for. RESULTS: Incidence of ischaemic findings, defined as Minnesota codes I, IV(1-3), V(1-3) (at rest and after the load), or receiving treatment for ischaemia, was significantly higher in the exposed workers, especially for the spinning/refining workers (adjusted OR 2.1; 95% CI 1.1 to 4.0) or the highest quartile of six year mean TTCA (adjusted OR 3.9; 95% CI 1.8 to 8.7), although the observed increase in risk was diminished when rigorous ECG criteria were applied. Incidence of retinal microaneurysm was increased with marginal significance. Among cardiovascular risk factors we examined, only blood pressure values were significantly increased in the exposed workers. CONCLUSIONS: Increased risk of ischaemic electrocardiogram findings among Japanese viscose rayon workers was observed. Although its clinical significance is to be discussed, the current Japanese occupational exposure limit for CS2, 10 ppm, would be high to prevent subclinical cardiovascular effects in this study population. PMID- 14739379 TI - Accuracy of task recall for epidemiological exposure assessment to construction noise. AB - AIMS: To validate the accuracy of construction worker recall of task and environment based information; and to evaluate the effect of task recall on estimates of noise exposure. METHODS: A cohort of 25 construction workers recorded tasks daily and had dosimetry measurements weekly for six weeks. Worker recall of tasks reported on the daily activity cards was validated with research observations and compared directly to task recall at a six month interview. RESULTS: The mean L(EQ) noise exposure level (dBA) from dosimeter measurements was 89.9 (n = 61) and 83.3 (n = 47) for carpenters and electricians, respectively. The percentage time at tasks reported during the interview was compared to that calculated from daily activity cards; only 2/22 tasks were different at the nominal 5% significance level. The accuracy, based on bias and precision, of percentage time reported for tasks from the interview was 53-100% (median 91%). For carpenters, the difference in noise estimates derived from activity cards (mean 91.9 dBA) was not different from those derived from the questionnaire (mean 91.7 dBA). This trend held for electricians as well. For all subjects, noise estimates derived from the activity card and the questionnaire were strongly correlated with dosimetry measurements. The average difference between the noise estimate derived from the questionnaire and dosimetry measurements was 2.0 dBA, and was independent of the actual exposure level. CONCLUSIONS: Six months after tasks were performed, construction workers were able to accurately recall the percentage time they spent at various tasks. Estimates of noise exposure based on long term recall (questionnaire) were no different from estimates derived from daily activity cards and were strongly correlated with dosimetry measurements, overestimating the level on average by 2.0 dBA. PMID- 14739380 TI - Psychosocial work environment and indoor air problems: a questionnaire as a means of problem diagnosis. AB - AIMS: To examine the relation between the psychosocial work environment and the perceived indoor air problems measured by a questionnaire survey; and to discuss the role of a questionnaire as a means to enhance collaboration in the challenging multiprofessional process of solving indoor air problems. METHODS: The research material comprises surveys conducted in 1996-99 in 122 office workplaces with 11 154 employees. RESULTS: The association between the psychosocial work environment measured by the Indoor Air Questionnaire (MM-40) and the occupants' complaints concerning indoor air as well as symptoms attributed to indoor air was significant. Those who perceived their psychosocial work environment more negatively had more complaints regarding the indoor environment and more symptoms attributed to the indoor air. The association was detected among both genders, in every age group, among smokers and non-smokers, and respondents with an allergic or a non-allergic background. CONCLUSIONS: Results support the hypothesis that psychosocial factors in the work environment play a significant role in indoor air problems at workplaces. The survey data can be used as a reference database for future studies, and in occupational health care practice when the working conditions of individual workplaces are estimated. The MM-40 could be useful as a practical screening method in field work for analysing the role of the psychosocial work environment among the different background factors of an indoor air problem. However, in order to interpret and evaluate the significance of the results concerning a single workplace, more information on the organisation is needed, as well as cooperation and discussions with the staff. Further studies of the reliability and validity of the psychosocial questions in MM-40 are also needed. PMID- 14739381 TI - Update of a prospective study of mortality and cancer incidence in the Australian petroleum industry. AB - AIMS: To update the analysis of the cohort mortality and cancer incidence study of employees in the Australian petroleum industry. METHODS: Employees from 1981 to 1996 were traced through the Australian National Death Index and the National Cancer Statistics Clearing House. Cause specific mortality and cancer incidence were compared with those of the Australian population by means of standardised mortality ratios (SMRs) and standardised incidence ratios (SIRs). Associations between increased incidence of specific cancers and employment in the petroleum industry were tested by trends according to period of first employment, duration of employment, latency, and hydrocarbon exposure, adjusting for personal smoking history where appropriate. Total follow up time was 176 598 person-years for males and 10 253 person-years for females. RESULTS: A total of 692 of the 15 957 male subjects, and 16 of the 1206 female subjects had died by the cut off date, 31 December 1996. In males, the all-cause SMR and the SMRs for all major disease categories were significantly below unity. There was a non-significant increase of the all-cancer SIR (1.04, 95% CI 0.97 to 1.11). There was a significant increase of the incidence of melanoma (SIR 1.54, 95% CI 1.30 to 1.81), bladder cancer (SIR 1.37, 95% CI 1.00 to 1.83), and prostate cancer (SIR 1.19, 95% CI 1.00 to 1.40), and a marginally significant excess of pleural mesothelioma (SIR 1.80, 95% CI 0.90 to 3.22), leukaemia (SIR 1.39, 95%CI 0.91 to 2.02), and multiple myeloma (SIR 1.72, 95% CI 0.96 to 2.84). CONCLUSIONS: Most cases of mesothelioma are probably related to past exposure to asbestos in refineries. The melanoma excess may be the result of early diagnosis. The excess bladder cancer has not been observed previously in this industry and is not readily explained. The divergence between cancer incidence and cancer mortality suggests that the "healthy worker effect" may be related to early reporting of curable cancers, leading to increased likelihood of cure and prolonged mean survival time. PMID- 14739382 TI - Lung cancer and dust exposure: results of a prospective cohort study following 3260 workers for 50 years. AB - AIMS: To study the lasting health impact of occupational dust exposure on life expectancy and specific causes of death. METHODS: Male Viennese workers, selected at age > or =40 (mean 54) years during preventive check-ups between 1950 and 1960, were followed prospectively until death. Half of them (1630) were exposed at work to (non-fibrous) particulates, while the non-exposed workers were matched for year, age, and smoking status at the start of observation. RESULTS: Average life expectancy of those exposed was 1.6 years less than that of those non exposed. Only a small part of this decrease in life expectancy (hazards ratios in brackets) was related to acknowledged occupational diseases such as silicosis and silicotuberculosis (67.12). Chronic obstructive lung disease (1.82) and cancer of the lung (1.42) and stomach (1.77) were found more frequently among those exposed. CONCLUSIONS: Results support the hypothesis that high exposure to insoluble particulates such as silica in the metal, glass, ceramics, and stone industries promotes bronchial cancer and chronic obstructive pulmonary disease. The finding of an increased incidence of stomach cancer might be related to particles swallowed after clearance from the airways. PMID- 14739383 TI - Integrating qualitative methods into occupational health research: a study of women flight attendants. AB - AIMS: To identify possible work related sources of psychosocial stress in order to develop pertinent questionnaire items for a cross-sectional health survey of 3000 Italian women flight attendants, and to solicit suggestions on improving survey participation. METHODS: Qualitative study in which 26 current and former women flight attendants participated: three focus groups of 6-7 participants each and six in-depth individual interviews. The session themes included positive and negative aspects of the job; relationships with colleagues, superiors, and passengers; perception of occupational risk for serious diseases; compatibility of work and family; and experiences of work related sources of stress and their effect on health. A transcript based analysis of the focus groups and interviews was used to identify emerging themes related to risk factors for mental health problems. RESULTS: The participants indicated that mental health was a major concern. Several work related risk factors possibly related to adverse outcomes, such as depression and anxiety, were highlighted. These included isolation and solitude, fears of being inadequate partners and mothers due to job demands, passenger relationships, and lack of protection by employers with respect to workplace exposures and violent passengers. The information gained was used to develop a mental health module for inclusion in the health survey questionnaire which included questions on history of severe depression or anxiety, suicidal ideation or attempt, substance abuse, workplace sexual harassment, social support, leisure time activities, relationship with a partner, and role as mother. CONCLUSIONS: Employing qualitative methods to identify work related sources of psychosocial stress enabled development of pertinent questionnaire items for a cross-sectional epidemiological study of women flight attendants. Follow up qualitative research may be necessary in order to put the cross sectional study findings into context and to explore actions or strategies for preventing work related health problems evidenced from the survey. PMID- 14739384 TI - Attention and working memory in resident anaesthetists after night duty: group and individual effects. AB - AIMS: To investigate the effects of a single period of night duty on measures of attention and working memory in a group of residents (registrars) in anaesthesiology. Emphasis was placed on individual deficits using a reference point of the equivalent effect of a blood alcohol concentration (BAC) >0.05% determined by other researchers. METHODS: There were 33 subjects aged 26-42 years. Night duty was performed on a weekly basis. Baseline assessments were conducted at either 08 15 or 08 55 preceding night duty and repeated 24-25 hours later, just after the completion of duty. Questionnaires included items regarding duration of sleep and the Stanford Sleepiness Scale. A battery of four reaction time (RT) tasks of increasing difficulty, lasting approximately 35 minutes, was administered on a personal computer. These ranged from simple RT to progressively more complex RT tasks incorporating working memory. A significant change was regarded as >15% deterioration in respect of speed or accuracy. RESULTS: The mean duration of sleep preceding night duty was 7.04 hours and 1.66 hours during the period of night duty. Intergroup comparisons revealed significant prolongation in mean response speed in the first three tests. Mean accuracy was significantly reduced only in respect of the two more complex tests. A >15% deterioration in response speed occurred in up to 30% of subjects on a single task, rising to 52% (17/33) overall. Deterioration occurred in a patchy distribution in most subjects, involving no more than one or two of the four tasks. As regards accuracy, the prevalence of deterioration increased with task complexity. CONCLUSIONS: Results are in general agreement with previous group analyses. A new dimension was added by the analysis of a broad spectrum of individual response to sleep deprivation. The effects of sleep loss in residents cannot be overlooked, even in a relatively benign work schedule. PMID- 14739385 TI - Incidence of cancer among Swedish military and civil personnel involved in UN missions in the Balkans 1989-99. AB - Leukaemia cases among European UN soldiers in the Balkans have been related hypothetically to exposure to depleted uranium. This study was performed to investigate the risk of cancer among Swedish personnel (8750 men and 438 women) involved in UN missions in the Balkans 1989-99. The overall incidence of cancer was slightly higher than expected; 34 cancers were observed and 28.1 were expected based on national cancer rates. Among military men, there were eight cases of testicular cancer versus 4.6 expected. There was one case of chronic myeloid leukaemia, and no cases of acute leukaemia. The overall risk of cancer was increased in a subgroup of 648 men taking part in convoy operations, based on only five cancers at four different sites. The study gives no support for the hypothesis that UN service in the Balkans could lead to haematolymphatic malignancies after short latency. However, no exposure assessment was performed, and future follow up is necessary for evaluation of long term risks. PMID- 14739386 TI - Risk of selected birth defects by maternal residence close to power lines during pregnancy. AB - AIMS: To evaluate selected birth outcomes from a published Norwegian cohort study in a nested case-control design with improved exposure data. METHODS: Two controls matched for sex, year of birth, and municipality were selected randomly for children with the following defects: central nervous system (CNS) defects, cardiac defects, respiratory system defects, oesophageal defects, and clubfoot. The distances between maternal addresses, during pregnancy, and power lines were obtained from maps mainly of scale 1:5000. The magnetic fields in the residences were estimated based on distance, current, voltage, and configuration. RESULTS: The highest increased risks were seen for hydrocephalus (OR 1.73, 95% CI 0.26 to 11.64) and for cardiac defects (OR 1.54, 95% CI 0.89 to 2.68). CONCLUSION: This study does not support the hypothesis that residential exposure to electromagnetic fields from power lines causes any of the investigated outcomes. PMID- 14739387 TI - Temporal and seasonal variation in the occurrence of chemical incidents. AB - AIMS: To determine whether there were temporal or seasonal patterns in the occurrence of chemical incidents reported to the West Midlands Chemical Incident Surveillance System, UK. METHODS: A retrospective analysis of reports maintained on a computerised database was carried out for information received from January 1997 to December 2001. RESULTS: Annual numbers of events increased significantly from 77 (7.5%) in 1997 to 282 (27.4%) in 2001. Compared to the average of other seasons, proportionately more incidents occurred in the summer (29.3%). Compared with the average of other days, incidents were more likely to occur on Thursdays (17.1%) and least likely on Saturdays (8.9%). When grouped together on a six hourly basis incidents were most frequent between 12 00 and 17 59 (31.1%) and least frequent between 00 00 and 05 59 (21.2%). CONCLUSIONS: The surveillance system shows that chemical incidents do not occur randomly but have marked temporal and seasonal variation. These results have implications for service provision, training, and the development of preventive strategies. PMID- 14739388 TI - World at work: the electronics industry. PMID- 14739389 TI - Inhaling volcanic ash on Montserrat. PMID- 14739390 TI - Sick building syndrome. PMID- 14739391 TI - The association between sex segregation, working conditions, and sickness absence among employed women. AB - AIMS: To analyse the association between sickness absence and sex segregation of occupation and of work site, respectively, and to analyse work environmental factors associated with high sickness absence. METHODS: The study group consisted of 1075 women employed as nurses, assistant nurses, medical secretaries, or metal workers who answered a questionnaire comprising 218 questions on women's health and living conditions. Sickness absence was collected from employers' and social insurance registers. RESULTS: Women working in the male dominated occupation had in general higher sickness absence compared to those working in female dominated occupations. However, metal workers at female dominated work sites had 2.98 (95% CI 2.17 to 3.79) sick-leave spells per woman and year compared to 1.70 (95% CI 1.29 to 2.10) among those working with almost only men. In spite of a better physical work environment, female metal workers at a female dominated work site had a higher sickness absence than other women, which probably could be explained by the worse psychosocial work environment. Working with more women also had a positive association to increased frequency of sick-leave spells in a multivariate analysis including several known indicators of increased sick-leave. CONCLUSIONS: There was an association between sickness absence and sex segregation, in different directions at the occupational and work site level. The mechanism behind this needs to be more closely understood regarding selection in and out of an occupation and a certain work site. PMID- 14739392 TI - The inhibition of mercury absorption by dietary ethanol in humans: cross sectional and case-control studies. AB - BACKGROUND: Since the inhibition of mercury absorption by ethanol was serendipitously discovered in 1965,(1) a limited number of small number studies with both animal and human subjects have reported results consistent with this finding. AIMS: To investigate this phenomenon in a large scale human study with low level Hg exposed dentists. METHODS: Data were collected for a sample of 1171 dentists, and both cross sectional and case-control methods were utilised to examine the data. RESULTS: Abstainers (n = 345) had significantly higher urinary mercury concentrations (HgU) than drinkers (n = 826): 5.4 microg/l v 4.8 microg/l. Multiple linear regression showed a significant effect of ethanol dose on HgU after adjusting for potential confounders. A case-control analysis in which cases were defined as those individuals with urinary Hg concentrations of > or =15 microg/l (approximately top 5%), and controls as those with concentrations of <1.0 microg/l ( approximately bottom 5%), showed a clear protective dose response relation; there was a decreasing risk of being a "case" (having an HgU > or =15 microg/l) with increasing ethanol consumption. The significance of the adjusted model is p<0.001, and the chi2 test for trend across ethanol consumption categories in the adjusted model is p<0.05, confirming the dose-response relation. CONCLUSION: We believe that this straightforward investigation provides the first specific confirmation in a large scale human study of the inhibitory effect of ethanol on urinary mercury concentration, and by inference, on mercury absorption. PMID- 14739393 TI - A mortality cohort study among workers in a graphite electrode production plant in Italy. AB - BACKGROUND: Graphite electrode manufacturing workers are exposed to coal tar and its volatiles containing a variety of polycyclic aromatic hydrocarbons (PAH), silica and graphite dusts, and asbestos. AIMS: To investigate mortality from cancer and other diseases among workers in a graphite electrode production plant in Italy. METHODS: A total of 1291 males actively employed between 1 January 1950 and 31 December 1989 who had worked at the plant for at least one year were studied. The follow up extended from 1950 to 1997. Standardised mortality ratios (SMR) and their 95% confidence intervals (CI) were computed using mortality rates for the Italian and regional male population. RESULTS: Excess mortality was observed for all causes (SMR 1.44, CI 1.32 to 1.56), all cancers (SMR 1.27, CI 1.07 to 1.50), liver cancer (SMR 4.19, CI 2.68 to 6.23), silicosis (SMR 66.39, CI 52.56 to 82.7), and cirrhosis and other chronic diseases of the liver (SMR 1.87, CI 1.31 to 2.59) in comparison with the national male population. When regional rates were used to calculate the number of expected deaths, SMRs remained higher for silicosis (SMR 57.32, 42.11 to 76.22), and liver cancer (SMR 2.57, 1.57 to 3.97). Mortality from silicosis was increased in workers hired at young ages (<25 years, SMR 81.79; 25-34 years, SMR 82.73), and in workers aged <45 at death (SMR 333.3, CI 159.8 to 613). Mortality from liver cancer increased threefold (SMR 3.11, CI 1.78 to 5.05) in workers with more than 10 years of employment at the plant during the manufacture of Karbate products. CONCLUSIONS: Results support the association between excess mortality from silicosis and occupational exposure to siliceous sands experienced during graphite electrode manufacturing. The observed excess mortality from liver cancer is compatible, to some extent, with exposures that may have occurred during the manufacture of phenolic and furfuryl resins treated products, although a role of lifestyle factors and viral infections cannot be excluded. PMID- 14739394 TI - The physiological regulation of thirst and fluid intake. AB - Thirst is important for maintaining body fluid homeostasis and may arise from deficits in either intracellular or extracellular fluid volume. Neural signals arising from osmotic and hormonal influences on the lamina terminalis may be integrated within the brain, with afferent information relayed from intrathoracic baroreceptors via the hindbrain to generate thirst. PMID- 14739395 TI - Why do we not all have proteinuria? An update of our current understanding of the glomerular barrier. AB - The key question is not why some patients have proteinuria but rather why not all people have it. In the present review, we will present an update on the glomerular barrier after the recent breakthroughs in podocyte biology. In particular, we will discuss the role of the endothelium, which seems to be a neglected part of the glomerular membrane. PMID- 14739396 TI - On heat and cells and proteins. AB - Two principal forms of temperature-control strategies have evolved, i.e., poikilothermic and homeothermic life. Even in homeothermic animals, the temperature field of the body is not homogeneous. These observed temperature differences can affect cellular function directly or via the expression of heat shock or cold shock proteins. PMID- 14739398 TI - Selectins facilitate carcinoma metastasis and heparin can prevent them. AB - Selectins are cell adhesion molecules mediating attachment of leukocytes to activated endothelium as well as the adhesion reaction of tumors during malignancy. Heparin, which is known to attenuate metastasis, is a potent blocker of selectins. Here, the role of selectins in metastasis and the potential of heparin to modulate malignancy are discussed. PMID- 14739399 TI - Hypothalamic and midbrain circuitry that distinguishes between escapable and inescapable pain. AB - Characteristics of emotional, behavioral, and physiological responses to pain are determined to a large extent by the behavioral significance of the pain, in particular to the degree to which the pain can be escaped. This review presents evidence that these different patterns of response depend on the activation of distinct pathways within the brain. PMID- 14739400 TI - Tracking the moveable feast: sonomicrometry and gastrointestinal motility. AB - Ultrasonomicrometry measures distance between piezoelectric crystals based on transmission time of ultrasound bursts. It allows monitoring of coordinated motion of small and delicate tissues, including gastrointestinal sphincters. Its suitability for motility studies in small animals such as mice suggests that its use in gastrointestinal studies will increase in coming years. PMID- 14739401 TI - Proteoglycans and brain repair. AB - Proteoglycans are complex molecules composed of long, unbranched sugar chains attached to a protein core. In the mammalian central nervous system, they are a major component of the extracellular matrix and of the cellular surface. After a central nervous system injury, their expression in the lesion area changes strongly and contributes to the inhibition of axon regrowth and brain repair. PMID- 14739402 TI - Cognitive deficits in hyperacute stroke. PMID- 14739403 TI - Stroke in china. PMID- 14739405 TI - Editorial comment--can MRI reliably detect hyperacute intracerebral hemorrhage? Ask the medical student. PMID- 14739406 TI - Parametric perfusion imaging with contrast-enhanced ultrasound in acute ischemic stroke. AB - BACKGROUND AND PURPOSE: Color-coded perfusion maps can be calculated from ultrasound harmonic gray-scale imaging data after ultrasound contrast agent bolus injection to analyze brain tissue perfusion. First reports indicate that this method can display cerebral perfusion deficits in acute ischemic stroke. We performed a prospective patient study to evaluate this approach. METHODS: Thirty consecutive patients suffering from acute middle cerebral artery infarction who presented to our department within 12 hours after symptom onset were investigated with ultrasound perfusion harmonic imaging (PHI) after Levovist bolus injection. Color-coded perfusion maps were calculated from the ultrasound data. In addition, the original gray-scale images were analyzed in cine mode. Findings were compared with those of cranial CT. RESULTS: All 30 patients suffered from acute ischemic stroke of the middle cerebral artery territory (median National Institutes of Health Stroke Scale score, 16 points). Twenty-three of the 30 patients (76.7%) had sufficient PHI insonation conditions. In 19 of these 23 patients (82.6%), a marked deficit in contrast enhancement could be visualized by initial PHI with the color-coded parameter images and cine-mode images. In 17 of the 23 (73.9%), the perfusion deficit was found on the parameter images. The area of hypoperfusion in the initial PHI investigation corresponded to the definite area of infarction in follow-up cranial CT. In 3 of 23 patients (13.0%), a perfusion deficit could be demonstrated in PHI, although the supplying artery was found patent by transcranial color-coded duplex sonography. CONCLUSIONS: With PHI, it is possible to display cerebral perfusion deficits in acute ischemic stroke. PHI yields additional information on the perfusion state of the human brain compared with extracranial and transcranial color-coded duplex sonography. PMID- 14739407 TI - NF-kappaB activation and Fas ligand overexpression in blood and plaques of patients with carotid atherosclerosis: potential implication in plaque instability. AB - BACKGROUND AND PURPOSE: Apoptosis is present in human atherosclerotic lesions. Nuclear factor-kappaB (NF-kappaB) is involved in the transcriptional regulation of the proapoptotic protein Fas ligand (FasL). We have analyzed NF-kappaB activation and FasL expression in atherosclerotic plaques and peripheral blood mononuclear cells (PBMCs) of patients with carotid stenosis. METHODS: NF-kappaB activation and FasL and active caspase-3 expression were analyzed in 32 human carotid plaques. NF-kappaB activation and FasL mRNA were tested in PBMCs of patients and healthy volunteers. We analyzed whether the NF-kappaB inhibitor parthenolide regulates FasL expression and cytotoxicity in human T cells. RESULTS: The inflammatory region of plaques showed an increase in NF-kappaB activation (3393+/-281 versus 1029+/-100 positive nuclei per mm(2), P<0.001) and FasL (16+/-1.4% versus 13+/-1.8%, P<0.05) and active caspase-3 (3.3+/-0.6 versus 1.5+/-0.3%, P<0.05) expression compared with the fibrous area. Activated NF kappaB and FasL protein were colocalized in plaque cells. In PBMCs obtained from those patients the day of endarterectomy, NF-kappaB activation and FasL expression were significantly increased compared with healthy controls (1.5+/-0.1 versus 0.5+/-0.1 and 2.1+/-0.1 versus 1.2+/-0.1 arbitrary units, respectively; P<0.001). There was a significant correlation between NF-kappaB activation and FasL expression. In activated T cells, parthenolide decreased NF-kappaB activation, FasL promoter activity, and mRNA expression. Parthenolide also decreased cytotoxicity of activated Jurkat cells on FasL-sensitive cells. CONCLUSIONS: NF-kappaB activation and FasL overexpression occur in PBMCs and atherosclerotic lesions of patients with carotid stenosis. The NF-kappaB-FasL pathway could be involved in the mechanisms underlying plaque instability in humans. PMID- 14739408 TI - Predictors of neurocardiogenic injury after subarachnoid hemorrhage. AB - BACKGROUND AND PURPOSE: Subarachnoid hemorrhage (SAH) frequently results in myocardial necrosis with release of cardiac enzymes. Historically, this necrosis has been attributed to coronary artery disease, coronary vasospasm, or oxygen supply-demand mismatch. Experimental evidence, however, indicates that excessive release of norepinephrine from the myocardial sympathetic nerves is the most likely cause. We hypothesized that myocardial necrosis after SAH is a neurally mediated process that is dependent on the severity of neurological injury. METHODS: Consecutive patients admitted with SAH were enrolled prospectively. Predictor variables reflecting demographic (age, sex, body surface area), hemodynamic (heart rate, systolic blood pressure), treatment (phenylephrine dose), and neurological (Hunt-Hess score) factors were recorded. Serial cardiac troponin I measurements and echocardiography were performed on days 1, 3, and 6 after enrollment. Troponin level was treated as a dichotomous outcome variable. We performed univariate and multivariate analyses on the relationships between the predictor variables and troponin level. RESULTS: The study included 223 patients with an average age of 54 years. Twenty percent of the subjects had troponin I levels >1.0 microg/L (range, 0.3 to 50 microg/L). By multivariate logistic regression, a Hunt-Hess score >2, female sex, larger body surface area and left ventricular mass, lower systolic blood pressure, and higher heart rate and phenylephrine dose were independent predictors of troponin elevation. CONCLUSIONS: The degree of neurological injury as measured by the Hunt-Hess grade is a strong, independent predictor of myocardial necrosis after SAH. This finding supports the hypothesis that cardiac injury after SAH is a neurally mediated process. PMID- 14739409 TI - Predictors of apparent diffusion coefficient normalization in stroke patients. AB - BACKGROUND AND PURPOSE: We sought to describe the frequency of normalization of apparent diffusion coefficient (ADC) values that are decreased in hyperacute stroke and to identify characteristics of tissue demonstrating normalization. METHODS: Sixty-eight acute ischemic stroke patients underwent MRI examination (including diffusion/perfusion imaging and MR angiography) within 6 hours (mean, 2.8 hours) after symptom onset, after 24 hours, and again 4 to 7 days later. Lesion volumes with decreased ADC and delayed time to peak in perfusion imaging were determined. In patients showing ADC normalization, volumes with ADC decrease graded as <50%, 50% to 60%, 60% to 70%, and 70% to 80% of the contralateral value were determined by thresholding. Patients were categorized as normalizers (demonstrating ADC normalization in >5 mL tissue with initially decreased ADC) or nonnormalizers (demonstrating ADC normalization in <5 mL tissue). RESULTS: Fourteen patients (19.7%) were classified as normalizers. Eleven of 31 patients (35.5%) initially imaged <3 hours after stroke onset and 3 of 37 (7.5%) of those imaged 3 to 6 hours after onset were normalizers. ADC normalization occurred predominantly in the basal ganglia and white matter after thrombolytic therapy in patients with more distal vessel occlusions. All normalizers demonstrated at least partial tissue reperfusion. Tissue with more severe initial decrease in ADC was less likely to demonstrate normalization. CONCLUSIONS: ADC normalization is not a rare event in acute stroke after tissue reperfusion. Brain tissue with initially decreased ADC, especially within 3 hours after stroke onset, may include "tissue at risk." PMID- 14739410 TI - Stroke magnetic resonance imaging is accurate in hyperacute intracerebral hemorrhage: a multicenter study on the validity of stroke imaging. AB - BACKGROUND AND PURPOSE: Although modern multisequence stroke MRI protocols are an emerging imaging routine for the diagnostic assessment of acute ischemic stroke, their sensitivity for intracerebral hemorrhage (ICH), the most important differential diagnosis, is still a matter of debate. We hypothesized that stroke MRI is accurate in the detection of ICH. To evaluate our hypotheses, we conducted a prospective multicenter trial. METHODS: Stroke MRI protocols of 6 university hospitals were standardized. Images from 62 ICH patients and 62 nonhemorrhagic stroke patients, all imaged within the first 6 hours after symptom onset (mean, 3 hours 18 minutes), were analyzed. For diagnosis of hemorrhage, CT served as the "gold standard." Three readers experienced in stroke imaging and 3 final-year medical students, unaware of clinical details, separately evaluated sets of diffusion-, T2-, and T2*-weighted images. The extent and phenomenology of the hemorrhage on MRI were assessed separately. RESULTS: Mean patient age was 65.5 years; median National Institutes of Health Stroke Scale score was 10. The experienced readers identified ICH with 100% sensitivity (confidence interval, 97.1 to 100) and 100% overall accuracy. Mean ICH size was 17.3 mL (range, 1 to 101.5 mL). The students reached a mean sensitivity of 95.16% (confidence interval, 90.32 to 98.39). CONCLUSIONS: Hyperacute ICH causes a characteristic imaging pattern on stroke MRI and is detectable with excellent accuracy. Even raters with limited film-reading experience reached good accuracy. Stroke MRI alone can rule out ICH and demonstrate the underlying pathology in hyperacute stroke. PMID- 14739411 TI - Magnetic resonance imaging improves detection of intracerebral hemorrhage over computed tomography after intra-arterial thrombolysis. AB - BACKGROUND AND PURPOSE: Unenhanced CT is routinely performed after intra-arterial (IA) thrombolysis. The presence of residual contrast causing staining of injured brain may mimic intracerebral hemorrhage (ICH). We evaluated MRI with diffusion weighted imaging (DWI) and susceptibility-weighted imaging (SWI) for detection of ICH after IA thrombolysis, specifically in equivocal areas of hyperdensity seen on postprocedure CT, to help guide the decision to use anticoagulation or antiplatelet therapy after the IA thrombolysis. METHODS: We performed a retrospective analysis of 15 consecutive patients who underwent IA thrombolysis for acute stroke between September 2000 and March 2003. Inclusion criteria required an immediate postprocedure CT with a questionable hyperdensity and, within the next 48 hours, an MRI with DWI and/or SWI. RESULTS: All patients had CT regions of hyperdensity that were equivocal for the presence of ICH. All patients subsequently underwent DWI, and 11 also underwent SWI. Eleven of 15 patients had magnetic susceptibility-induced hypointensity in DWI hyperintensity regions, signifying the presence of acute deoxyhemoglobin. Nine of these patients also received SWI, which confirmed the presence of blood within these regions. Follow-up CT on all 11 patients confirmed ICH. In the 4 patients without DWI susceptibility change, 0 were found to have ICH on either SWI (performed in 2 patients) or follow-up CT. MRI reliably detected the presence of ICH in all patients, whereas CT failed to differentiate contrast staining from hemorrhage in 4 of the 15 patients. CONCLUSIONS: MRI is an effective means to detect the presence of blood within an equivocal region on post-IA thrombolysis CT. This may influence the decision to use anticoagulation or antiplatelet therapy. PMID- 14739412 TI - Acute basilar artery occlusion: diffusion-perfusion MRI characterization of tissue salvage in patients receiving intra-arterial stroke therapies. AB - BACKGROUND AND PURPOSE: Diffusion-perfusion MRI in patients with anterior circulation occlusions has demonstrated salvage of threatened tissue after thrombolytic therapy. Similar studies have not been reported with posterior circulation occlusions. METHODS: Patients with acute basilar artery occlusion treated with intra-arterial thrombolytics were studied with multimodal MRI before treatment, several hours after treatment, and at day 7. RESULTS: Ten patients were studied (9 men, 1 woman). Mean age was 70 years, and median pretreatment National Institutes of Health Stroke Scale (NIHSS) score was 14. In 6 patients imaged before treatment and at day 7, mean pretreatment diffusion-weighted imaging (DWI) lesion volume was 11 cm(3), and day 7, lesion volume was 2.6 cm(3). Significant mismatch was visualized in all 5 patients with pretreatment perfusion diffusion imaging (mean, 73%; range, 49% to 99%). Late imaging obtained in 4 of these 5 patients demonstrated that mean posttreatment DWI lesion volume (21 cm(3)) was less than the mean initial perfusion lesion volume (62 cm(3)). Although there was no direct correlation between pretreatment DWI volume and initial NIHSS (r=-0.113), there was good correlation between pretreatment perfusion-weighted imaging volume and initial NIHSS (r=0.72). CONCLUSIONS: In this first report of diffusion-perfusion MRI in patients with acute basilar artery occlusions treated with intra-arterial thrombolysis, significant mismatch was visualized on pretreatment studies, suggesting that large volumes of salvageable tissue were present. Final infarct volumes were smaller than pretreatment perfusion volumes, suggesting that substantial volumes of tissue were salvaged by thrombolytic reperfusion. PMID- 14739413 TI - Neuroendoscopic management of intraventricular hemorrhage. AB - BACKGROUND AND PURPOSE: We reviewed our 7-year experience in neuroendoscopic management of severe intraventricular hemorrhage (IVH) to evaluate its safety, efficiency, and efficacy. METHODS: Thirteen patients with spontaneous primary or secondary tetraventricular IVH underwent neuroendoscopy. In all procedures, we used a flexible instrument. CT scans obtained before and after surgery were compared for Graeb score and ventriculocranial ratio. Glasgow Outcome Scale was assessed at 12 months. RESULTS: In all patients, the procedure resulted in a substantial removal of ventricular blood. Graeb score was reduced by 65%, and ventriculocranial ratio was reduced by 30% (P<0.002). The procedure was carried out safely even in the presence of a vascular malformation, and no rebleeding or delayed hydrocephalus was observed in any case. Mortality at 12 months was 30.7%. Favorable outcome (Glasgow Outcome Scale, 3 to 5) was observed in 61.5% of cases. CONCLUSIONS: Neuroendoscopic management of severe IVH in this cohort of patients was safe, efficiently reduced the amount of ventricular blood and ventricular dilatation, and effectively produced an outcome profile that compares very favorably with other more conventional treatments. PMID- 14739414 TI - Uncomplicated rapid posthypothermic rewarming alters cerebrovascular responsiveness. AB - BACKGROUND AND PURPOSE: Recently, we focused on the cerebrovascular protective effects of moderate hypothermia after traumatic brain injury, noting that the efficacy of posttraumatic hypothermia is related to the rate of posthypothermic rewarming. In the current communication, we revisit the use of hypothermia with varying degrees of rewarming to ascertain whether, in the normal cerebral vasculature, varying rates of rewarming can differentially affect cerebrovascular responsiveness. METHODS: Pentobarbital-anesthetized rats equipped with a cranial window were randomized to 3 groups. In 1 group, a 1-hour period of hypothermia (32 degrees C) followed by slow rewarming (over 90 minutes) was used. In the remaining 2 groups, either a 1- or 2-hour period of hypothermia was followed by rapid rewarming (within 30 minutes). Vasoreactivity to hypercapnia and acetylcholine was assessed before, during, and after hypothermia. Additionally, the vascular responses to sodium nitroprusside (SNP) and pinacidil, a K(ATP) channel opener, were also examined. RESULTS: Hypothermia itself generated modest vasodilation and reduced vasoreactivity to all utilized agents. The slow rewarming group showed restoration of normal vascular responsivity. In contrast, hypothermia followed by rapid rewarming was associated with continued impaired responsiveness to acetylcholine and arterial hypercapnia. These abnormalities persisted even with the use of more prolonged (2-hour) hypothermia. Furthermore, posthypothermic rapid rewarming impaired the dilator responses of SNP and pinacidil. CONCLUSIONS: Posthypothermic rapid rewarming caused cerebral vascular abnormalities, including a diminished response to acetylcholine, hypercapnia, pinacidil, and SNP. Our data with acetylcholine and SNP suggest that rapid rewarming most likely causes abnormality at both the vascular smooth muscle and endothelial levels. PMID- 14739415 TI - Noninvasive quantification of brain edema and the space-occupying effect in rat stroke models using magnetic resonance imaging. AB - BACKGROUND AND PURPOSE: Brain edema is a life-threatening consequence of stroke and leads to an extension of the affected tissue. The space-occupying effect due to brain edema can be quantified in rat stroke models with the use of MRI. The present study was performed to test 2 hypotheses: (1) Can quantification of the space-occupying effect due to brain edema serve as a noninvasive measure for brain water content? (2) Does morphometric assessment of brain swelling allow determination of true infarct size on MRI after correction for the space occupying effect of edema? METHODS: Thirty rats were subjected to permanent suture middle cerebral artery occlusion. MRI was performed after 6 or 24 hours, and hemispheric swelling was assessed morphometrically. Interobserver and intraobserver agreements were determined for MRI measurements. In study I, the space-occupying effect due to brain edema was correlated with the absolute brain water content by the wet/dry method. In study II, lesion volumes corrected and uncorrected for edema were calculated on MRI and on TTC staining and compared. RESULTS: Interobserver and intraobserver agreements for MRI measurements were excellent (r>or=0.97). Brain water content and hemispheric swelling correlated well after 6 and 24 hours (r>or=0.95). Corrected lesion volumes correlated with r=0.78 between TTC staining and MRI. Without edema correction, lesion volumes were overestimated by 20.3% after 6 hours and by 29.6% after 24 hours of ischemia. CONCLUSIONS: Morphometric assessment of hemispheric swelling on MRI can determine the increase in absolute brain water content noninvasively and can also provide ischemic lesion volumes corrected for brain edema. PMID- 14739416 TI - Increased NADPH-oxidase activity and Nox4 expression during chronic hypertension is associated with enhanced cerebral vasodilatation to NADPH in vivo. AB - BACKGROUND AND PURPOSE: We examined the importance of NADPH-oxidase in reactive oxygen species production in cerebral arteries and its effect on vascular tone in vivo. Furthermore, we investigated whether chronic hypertension affects function or expression of this enzyme in cerebral vessels. METHODS: Superoxide generation was detected in isolated rat basilar arteries with the use of lucigenin-enhanced chemiluminescence. mRNA expression of NADPH-oxidase subunits was assessed by real time polymerase chain reaction. Basilar artery diameter was measured with the use of a cranial window preparation in anesthetized rats. RESULTS: NADPH-stimulated superoxide production was 2.3-fold higher in arteries from spontaneously hypertensive rats (SHR) versus normotensive Wistar-Kyoto rats (WKY) and could be blocked by the NADPH-oxidase inhibitor diphenyleneiodonium. Higher NADPH-oxidase activity was also reflected at the molecular level as mRNA expression of the NADPH-oxidase subunit Nox4 was 4.1-fold higher in basilar arteries from SHR versus WKY. In contrast, expression of Nox1, gp91phox, p22phox, and p47phox did not differ between strains. Application of NADPH to basilar arteries caused larger vasodilatation in SHR than WKY. Vasodilatation to NADPH could be attenuated by diphenyleneiodonium, as well as diethyldithiocarbamate (Cu(2+)/Zn(2+)-superoxide dismutase inhibitor), catalase (H(2)O(2) scavenger), or tetraethylammonium (BK(Ca) channel inhibitor). CONCLUSIONS: Activation of NADPH oxidase in cerebral arteries generates superoxide, which is dismutated by Cu(2+)/Zn(2+)-superoxide dismutase to H(2)O(2). H(2)O(2) then elicits vasodilatation via activation of BK(Ca) channels. Upregulation of Nox4 during chronic hypertension is associated with elevated cerebral artery NADPH-oxidase activity. PMID- 14739417 TI - Antithrombotic drugs for carotid artery dissection. PMID- 14739418 TI - Functional magnetic resonance imaging before and after aphasia therapy: shifts in hemodynamic time to peak during an overt language task. AB - BACKGROUND AND PURPOSE: Comparing the temporal characteristics of hemodynamic responses in activated cortical regions of aphasic patients before and after therapy would provide insight into the relationship between improved task performance and changes in blood oxygenation level-dependent (BOLD) functional MRI (fMRI) signal. This study investigated differences in the time to peak (TTP) of hemodynamic responses in activated regions of interest (ROIs), before and after therapy, and related them to changes in task performance. METHODS: Three aphasic patients and 3 controls overtly generated a single exemplar in response to a category. For the patients, TTP of hemodynamic responses in selected ROIs was compared before and after language therapy. The timing differences between auditory cues and verbal responses were compared with TTP differences between auditory and motor cortices. RESULTS: The selected ROIs were significantly activated in both aphasic patients and controls during overt word generation. In the aphasic patients, both the timing difference from auditory cues to verbal responses and the TTP difference between auditory and motor cortices decreased after rehabilitation, becoming similar to the values found in controls. CONCLUSIONS: Findings indicate that (1) rehabilitation increased the speed of word-finding processes; (2) TTP analysis was sensitive to this functional change and can be used to represent improvement in behavior; and (3) it is important to monitor the behavioral performance that might correlate with the temporal pattern of the hemodynamic response. PMID- 14739419 TI - Sex differences in carotid plaque and stenosis. AB - BACKGROUND AND PURPOSE: Women are relatively protected from cardiovascular events; they are 3 times as likely as men to survive to age 90 years. Although clinical trials show an excess of thrombotic events with estrogen/progestin hormone replacement therapy, much experimental and epidemiological evidence suggests that estrogen may have beneficial effects on endothelial function and atherosclerosis, raising the possibility of sex differences in arterial remodeling. We studied sex differences in carotid plaque and stenosis in relation to survival free of stroke, death, and myocardial infarction. METHODS: A total of 1686 patients from an atherosclerosis prevention clinic were followed annually for up to 5 years (mean, 2.5+/-1.3 years) with baseline and follow-up measurements; there were 45 strokes, 94 myocardial infarctions, and 41 deaths. RESULTS: Carotid stenosis and plaque increased with age. Women had greater stenosis compared with men (P=0.001), whereas men had greater plaque area than did women at all ages (P<0.0001). Stroke, myocardial infarction, and death combined were predicted significantly by plaque area (P=0.004) but not by stenosis (P=0.042). CONCLUSIONS: Women have more stenosis but less plaque than men, suggesting that differences in sex hormones may affect remodeling of atherosclerosis. Plaque area was a stronger predictor of outcomes than was stenosis. PMID- 14739420 TI - Collagen type I alpha2 (COL1A2) is the susceptible gene for intracranial aneurysms. AB - BACKGROUND AND PURPOSE: The collagen alpha2(I) gene (COL1A2) on chromosome 7q22.1, a positional and functional candidate for intracranial aneurysm (IA), was extensively screened for susceptibility in Japanese IA patients. METHODS: Twenty one single nucleotide polymorphisms (SNPs) of COL1A2 were genotyped in genomic DNA from 260 IA patients (including 115 familial cases) (mean age, 59.9 years) and 293 controls (mean age, 61.6 years). Differences in allelic and genotypic frequencies between the patients and controls were evaluated with the chi(2) test. Circular dichroism spectrometry was monitored with collagen-related peptides that mimic triple-helical models of type I collagen with Ala-459 and Pro 459 to estimate the conformation and stability of alterations. RESULTS: Significant genotypic association in the dominant model was observed between an exonic SNP of COL1A2 and familial IA patients (chi(2)=11.08; df=1; P=0.00087; odds ratio=3.19; 95% CI, 2.22 to 6.50). This SNP induces Ala to Pro substitution at amino acid 459, located on a triple-helical domain. Circular dichroism spectra showed that the Pro-459 peptide had a higher thermal stability than the Ala-459 peptide. CONCLUSIONS: The variant of COL1A2 could be a genetic risk factor for IA patients with family history. PMID- 14739421 TI - Benefit of clopidogrel over aspirin is amplified in patients with a history of ischemic events. AB - BACKGROUND AND PURPOSE: The goal of this study was to examine the influence of preexisting symptomatic atherosclerotic disease on subsequent ischemic event rates and compare the efficacy of clopidogrel versus aspirin (acetylsalicylic acid, ASA) in patients with such disease. METHODS: Using the CAPRIE database, we performed multivariate analyses for patients who had symptomatic atherosclerotic disease (ischemic stroke [IS] or myocardial infarction [MI]) in their medical history before enrollment in the Clopidogrel Versus Aspirin in Patients at Risk of Ischemic Events (CAPRIE) trial. Two composite end points were used: (1) IS, MI, or vascular death and (2) IS, MI, or rehospitalization for ischemia. RESULTS: In the CAPRIE population, prior IS and MI each were statistically significant predictors of subsequent ischemic events. Compared with the overall population, patients with preexisting symptomatic atherosclerotic disease had elevated event rates for the end point of IS, MI, or vascular death; 3-year rates were 20.4% with clopidogrel and 23.8% with ASA (absolute risk reduction, 3.4%; 95% CI, -0.2 to 7.0; number needed to treat, 29; relative risk reduction, 14.9%; P=0.045). Similar results were obtained for the end point of IS, MI, or rehospitalization for ischemia; 3-year event rates were 32.7% with clopidogrel and 36.6% with ASA (absolute risk reduction, 3.9%; 95% CI, -0.4 to 8.1; number needed to treat, 26; relative risk reduction, 12.0%; P=0.039). CONCLUSIONS: CAPRIE patients with a history of prior symptomatic atherosclerotic disease had a high rate of subsequent ischemic events. The absolute benefit of clopidogrel over ASA seemed to be amplified in such high-risk patients. PMID- 14739422 TI - Crossed cerebellar diaschisis in patients with cortical infarction: logistic regression analysis to control for confounding effects. AB - BACKGROUND AND PURPOSE: Crossed cerebellar diaschisis (CCD) refers to reduced metabolism and blood flow in the cerebellar hemisphere contralateral to a cerebral lesion. Many cortical areas have been reported to cause CCD without consideration of confounding factors. We performed single-photon emission computed tomography (SPECT) in patients with cortical infarction to identify regions independently related to CCD, controlling for possible confounding effects. METHODS: Patients with unilateral cortical infarction (n=113; 75 male, 38 female; mean+/-SD age, 66+/-13 years) underwent SPECT of the brain with N isopropyl-p-[(123)I]iodoamphetamine ((123)I-IMP). Regional cerebral blood flow was measured autoradiographically. Asymmetry indices (AIs) were calculated on the basis of ratios representing symmetrical regional cerebral blood flow in the cerebellum and 16 cerebral regions. CCD was defined as AI for cerebellum >0.1. AIs for 16 cortical regions were considered for both dichotomous and continuous variables for analysis of CCD occurrence by means of backward logistic regression. RESULTS: For dichotomized variables, hypoperfusion of postcentral (odds ratio [OR]=7.607; 95% CI, 2.299 to 25.174) and supramarginal (OR=3.916; 95% CI, 1.394 to 11.003) regions independently influenced CCD. For continuous variables, hypoperfusion of postcentral (OR=1.044; 95% CI, 1.019 to 1.068) and supramarginal (OR=1.021; 95% CI, 1.001 to 1.041) regions (and, as a negative factor, medial occipital regions; OR=0.942; 95% CI, 0.895 to 0.991) independently influenced CCD. CONCLUSIONS: Many cortical areas apparently do not contribute to CCD. Correspondence of CCD between dichotomized and continuous analyses suggests that location of a lesion, not severity, is the main determinant of CCD. PMID- 14739424 TI - Global warming and nuclear power. PMID- 14739423 TI - Eligibility for recombinant tissue plasminogen activator in acute ischemic stroke: a population-based study. AB - BACKGROUND AND PURPOSE: Acute ischemic stroke patients are infrequently treated with recombinant tissue plasminogen activator (rtPA). We present unique population-based data regarding the eligibility of ischemic stroke patients for rtPA treatment. METHODS: All ischemic strokes presenting to an emergency department (ED) within a biracial population of 1.3 million were identified. The patient was considered eligible for rtPA on the basis of exclusion criteria from the National Institute of Neurological Disorders and Stroke rtPA trial. RESULTS: Of 2308 ischemic strokes, 1849 presented to an ED. Only 22% of all ischemic strokes in the population arrived in the ED in <3 hours from symptom onset; of these, 209 (51%) were ineligible for rtPA on the basis of mild stroke severity, medical and surgical history, or blood tests. CONCLUSIONS: In our population in 1993 to 1994, 8% of all ischemic stroke patients presented to an ED within 3 hours and met other eligibility criteria for rtPA. Even if time were not an exclusion for rtPA, only 29% of all ischemic strokes in our population would have otherwise been eligible for rtPA. PMID- 14739425 TI - Space exploration. Scientists add up gains, losses in Bush's new vision for NASA. PMID- 14739426 TI - Infectious diseases. Stopping Asia's avian flu: a worrisome third outbreak. PMID- 14739427 TI - Genetically modified organisms. Europe takes tentative steps toward approval of commercial GM crops. PMID- 14739428 TI - DOE national labs. University of California inches closer to preparing bids. PMID- 14739429 TI - Genetically modified organisms. Experts recommend a cautious approach. PMID- 14739430 TI - Biomedical politics. Democrats blast a sunny-side look at U.S. health disparities. PMID- 14739431 TI - Biomedical funding. NIH keeps grants staff but braces for change. PMID- 14739432 TI - Nonproliferation. North Korea's nuclear shell game. PMID- 14739433 TI - Genomics. New sequence boosts rats' research appeal. PMID- 14739434 TI - Arthur Carty profile. Canada's new science adviser hopes for good chemistry. PMID- 14739436 TI - American Astronomical Society meeting. Star formation--an extreme sport? PMID- 14739435 TI - American Astronomical Society meeting. Early galaxies baffle observers, but theorists shrug. PMID- 14739437 TI - American Astronomical Society meeting. A hot Jupiter sears its parent star. PMID- 14739438 TI - American Astronomical Society meeting. Snapshots from the meeting. PMID- 14739439 TI - Retraction of an interpretation. PMID- 14739440 TI - Brightening depression. PMID- 14739441 TI - The difficulties of testing for SARS. PMID- 14739442 TI - Comment on "Impact ejecta layer from the mid-Devonian: possible connection to global mass extinctions". PMID- 14739444 TI - Education. Bonus pay for research faculty. PMID- 14739445 TI - Cell biology. BAR domains go on a bender. PMID- 14739446 TI - Biochemistry. Mimicking posttranslational modifications of proteins. PMID- 14739447 TI - Materials science. Making nanoscale materials with supercritical fluids. PMID- 14739448 TI - Plant sciences. Imprinting--a green variation. PMID- 14739449 TI - Immunology. The Robin Hood of antigen presentation. PMID- 14739450 TI - Ferroelectricity at the nanoscale: local polarization in oxide thin films and heterostructures. AB - Ferroelectric oxide materials have offered a tantalizing potential for applications since the discovery of ferroelectric perovskites more than 50 years ago. Their switchable electric polarization is ideal for use in devices for memory storage and integrated microelectronics, but progress has long been hampered by difficulties in materials processing. Recent breakthroughs in the synthesis of complex oxides have brought the field to an entirely new level, in which complex artificial oxide structures can be realized with an atomic-level precision comparable to that well known for semiconductor heterostructures. Not only can the necessary high-quality ferroelectric films now be grown for new device capabilities, but ferroelectrics can be combined with other functional oxides, such as high-temperature superconductors and magnetic oxides, to create multifunctional materials and devices. Moreover, the shrinking of the relevant lengths to the nanoscale produces new physical phenomena. Real-space characterization and manipulation of the structure and properties at atomic scales involves new kinds of local probes and a key role for first-principles theory. PMID- 14739451 TI - Stable SNPs in malaria antigen genes in isolated populations. PMID- 14739452 TI - Detection of a red supergiant progenitor star of a type II-plateau supernova. AB - We present the discovery of a red supergiant star that exploded as supernova 2003gd in the nearby spiral galaxy M74. The Hubble Space Telescope (HST) and the Gemini Telescope imaged this galaxy 6 to 9 months before the supernova explosion, and subsequent HST images confirm the positional coincidence of the supernova with a single resolved star that is a red supergiant of 8(+4)(-2) solar masses. This confirms both stellar evolution models and supernova theories predicting that cool red supergiants are the immediate progenitor stars of type II-plateau supernovae. PMID- 14739453 TI - Phase diagram of degenerate exciton systems. AB - Degenerate exciton systems have been produced in quasi-two-dimensional confined areas in semiconductor coupled quantum well structures. We observed contractions of clouds containing tens of thousands of excitons within areas as small as (10 micron)2 near 10 kelvin. The spatial and energy distributions of optically active excitons were determined by measuring photoluminescence as a function of temperature and laser excitation and were used as thermodynamic quantities to construct the phase diagram of the exciton system, which demonstrates the existence of distinct phases. Understanding the formation mechanisms of these degenerate exciton systems can open new opportunities for the realization of Bose Einstein condensation in the solid state. PMID- 14739454 TI - Mesoporous silicates prepared using preorganized templates in supercritical fluids. AB - Well-ordered mesoporous silicate films were prepared by infusion and selective condensation of silicon alkoxides within microphase-separated block copolymer templates dilated with supercritical carbon dioxide. Confinement of metal oxide deposition to specific subdomains of the preorganized template yields high fidelity, three-dimensional replication of the copolymer morphology, enabling the preparation of structures with multiscale order in a process that closely resembles biomineralization. Ordered mesoporous silicate films were synthesized with dielectric constants as low as 1.8 and excellent mechanical properties. The films survive the chemical-mechanical polishing step required for device manufacturing. PMID- 14739455 TI - Electron transfer-induced dynamics of oxygen molecules on the TiO2(110) surface. AB - Diffusion of oxygen molecules on transition metal oxide surfaces plays a vital role for the understanding of catalysis and photocatalysis on these materials. By means of time-resolved scanning tunneling microscopy, we provide evidence for a charge transfer-induced diffusion mechanism for O2 molecules adsorbed on a rutile TiO2(110) surface. The O2 hopping rate depended on the number of surface donors (oxygen vacancies), which determines the density of conduction band electrons. These results may have implications for the understanding of oxidation processes on metal oxides in general. PMID- 14739456 TI - Viscosity of fluids in subduction zones. AB - The viscosities of aqueous fluids with 10 to 80 weight percent dissolved silicates have been measured at 600 degrees to 950 degrees C and 1.0 to 2.0 gigapascals by in situ observation of falling spheres in the diamond anvil cell. The viscosities at 800 degrees C range from 10(-4) to 10(0.5) pascal seconds. The combination of low viscosities with a favorable wetting angle makes silicate-rich fluid an efficient agent for material transport at low-volume fractions. Our results therefore suggest that there may be a direct relationship between the position of the volcanic front and the onset of complete miscibility between water and silicate melt in the subducting slab. PMID- 14739457 TI - Evidence that nitric acid increases relative humidity in low-temperature cirrus clouds. AB - In situ measurements of the relative humidity with respect to ice (RHi) and of nitric acid (HNO3) were made in both natural and contrail cirrus clouds in the upper troposphere. At temperatures lower than 202 kelvin, RHi values show a sharp increase to average values of over 130% in both cloud types. These enhanced RHi values are attributed to the presence of a new class of HNO3-containing ice particles (Delta-ice). We propose that surface HNO3 molecules prevent the ice/vapor system from reaching equilibrium by a mechanism similar to that of freezing point depression by antifreeze proteins. Delta-ice represents a new link between global climate and natural and anthropogenic nitrogen oxide emissions. Including Delta-ice in climate models will alter simulated cirrus properties and the distribution of upper tropospheric water vapor. PMID- 14739458 TI - T cell activation by lipopeptide antigens. AB - Unlike major histocompatibility proteins, which bind peptides, CD1 proteins display lipid antigens to T cells. Here, we report that CD1a presents a family of previously unknown lipopeptides from Mycobacterium tuberculosis, named didehydroxymycobactins because of their structural relation to mycobactin siderophores. T cell activation was mediated by the alphabeta T cell receptors and was specific for structure of the acyl and peptidic components of these antigens. These studies identify a means of intracellular pathogen detection and identify lipopeptides as a biochemical class of antigens for T cells, which, like conventional peptides, have a potential for marked structural diversity. PMID- 14739459 TI - Role of LBPA and Alix in multivesicular liposome formation and endosome organization. AB - What are the components that control the assembly of subcellular organelles in eukaryotic cells? Although membranes can clearly be distorted by cytosolic factors, very little is known about the intrinsic mechanisms that control the biogenesis, shape, and organization of organellar membranes. Here, we found that the unconventional phospholipid lysobisphosphatidic acid (LBPA) could induce the formation of multivesicular liposomes that resembled the multivesicular endosomes that exist where this lipid is found in vivo. This process depended on the same pH gradient that exists across endosome membranes in vivo and was selectively controlled by Alix. In turn, Alix regulated the organization of LBPA-containing endosomes in vivo. PMID- 14739460 TI - Snapshots of DsbA in action: detection of proteins in the process of oxidative folding. AB - DsbA, a thioredoxin superfamily member, introduces disulfide bonds into newly translocated proteins. This process is thought to occur via formation of mixed disulfide complexes between DsbA and its substrates. However, these complexes are difficult to detect, probably because of their short-lived nature. Here we show that it is possible to detect such covalent intermediates in vivo by a mutation in DsbA that alters cis proline-151. Further, this mutant allowed us to identify substrates of DsbA. Alteration of the cis proline, highly conserved among thioredoxin superfamily members, may be useful for the detection of substrates and intermediate complexes in other systems. PMID- 14739461 TI - Extensive gene traffic on the mammalian X chromosome. AB - Mammalian sex chromosomes have undergone profound changes since evolving from ancestral autosomes. By examining retroposed genes in the human and mouse genomes, we demonstrate that, during evolution, the mammalian X chromosome has generated and recruited a disproportionately high number of functional retroposed genes, whereas the autosomes experienced lower gene turnover. Most autosomal copies originating from X-linked genes exhibited testis-biased expression. Such export is incompatible with mutational bias and is likely driven by natural selection to attain male germline function. However, the excess recruitment is consistent with a combination of both natural selection and mutational bias. PMID- 14739462 TI - Multimodal signals: enhancement and constraint of song motor patterns by visual display. AB - Many birds perform visual signals during their learned songs, but little is known about the interrelationship between visual and vocal displays. We show here that male brown-headed cowbirds (Molothrus ater) synchronize the most elaborate wing movements of their display with atypically long silent periods in their song, potentially avoiding adverse biomechanical effects on sound production. Furthermore, expiratory effort for song is significantly reduced when cowbirds perform their wing display. These results show a close integration between vocal and visual displays and suggest that constraints and synergistic interactions between the motor patterns of multimodal signals influence the evolution of birdsong. PMID- 14739463 TI - The ubiquitin ligase SCFFbw7 antagonizes apoptotic JNK signaling. AB - Jun N-terminal kinases (JNKs) are essential for neuronal microtubule assembly and apoptosis. Phosphorylation of the activating protein 1 (AP1) transcription factor c-Jun, at multiple sites within its transactivation domain, is required for JNK induced neurotoxicity. We report that in neurons the stability of c-Jun is regulated by the E3 ligase SCF(Fbw7), which ubiquitinates phosphorylated c-Jun and facilitates c-Jun degradation. Fbw7 depletion resulted in accumulation of phosphorylated c-Jun, stimulation of AP1 activity, and neuronal apoptosis. SCF(Fbw7) therefore antagonizes the apoptotic c-Jun-dependent effector arm of JNK signaling, allowing neurons to tolerate potentially neurotoxic JNK activity. PMID- 14739464 TI - Human De-etiolated-1 regulates c-Jun by assembling a CUL4A ubiquitin ligase. AB - Arabidopsis thaliana De-etiolated-1 (AtDET1) is a highly conserved protein, with orthologs in vertebrate and invertebrate organisms. AtDET1 negatively regulates photomorphogenesis, but its biochemical mechanism and function in other species are unknown. We report that human DET1 (hDET1) promotes ubiquitination and degradation of the proto-oncogenic transcription factor c-Jun by assembling a multisubunit ubiquitin ligase containing DNA Damage Binding Protein-1 (DDB1), cullin 4A (CUL4A), Regulator of Cullins-1 (ROC1), and constitutively photomorphogenic-1. Ablation of any subunit by RNA interference stabilized c-Jun and increased c-Jun-activated transcription. These findings characterize a c-Jun ubiquitin ligase and define a specific function for hDET1 in mammalian cells. PMID- 14739465 TI - Selective differentiation of neural progenitor cells by high-epitope density nanofibers. AB - Neural progenitor cells were encapsulated in vitro within a three-dimensional network of nanofibers formed by self-assembly of peptide amphiphile molecules. The self-assembly is triggered by mixing cell suspensions in media with dilute aqueous solutions of the molecules, and cells survive the growth of the nanofibers around them. These nanofibers were designed to present to cells the neurite-promoting laminin epitope IKVAV at nearly van der Waals density. Relative to laminin or soluble peptide, the artificial nanofiber scaffold induced very rapid differentiation of cells into neurons, while discouraging the development of astrocytes. This rapid selective differentiation is linked to the amplification of bioactive epitope presentation to cells by the nanofibers. PMID- 14739466 TI - Long-term treatment of rheumatoid arthritis with tumour necrosis factor alpha blockade: outcome of ceasing and restarting biologicals. PMID- 14739467 TI - Anti-CD20 monoclonal antibody (rituximab) for refractory autoimmune thrombocytopenia in a girl with systemic lupus erythematosus. PMID- 14739468 TI - Polyarteritis nodosa as a presenting feature of angiomatoid fibrous histiocytoma. PMID- 14739469 TI - Leflunomide (Arava)-induced cystoid macular oedema. PMID- 14739470 TI - Sporothrix schenckii infection mimicking sarcoidosis. PMID- 14739471 TI - Terbinafine as a cause of cutaneous lupus erythematosus. PMID- 14739472 TI - Lower level of synovial fluid interferon-gamma in HLA-B27-positive than in HLA B27-negative patients with Chlamydia trachomatis reactive arthritis. PMID- 14739474 TI - Mononeuritis in Churg-Strauss syndrome in Asians responding to intravenous cyclophosphamide. PMID- 14739476 TI - Plasma endothelin-1 levels in chronic fatigue syndrome. PMID- 14739479 TI - Systemic juvenile idiopathic arthritis, Kikuchi's disease and haemophagocytic lymphohistiocytosis. PMID- 14739481 TI - Semiotics and fibromyalgia. PMID- 14739482 TI - Central nervous system involvement in perceived joint stiffness. PMID- 14739483 TI - Comment on 'Fibromyalgia and the therapeutic domain. A philosophical study on the origins of fibromyalgia in a specific social setting.' by Hazemeijer and Rasker. PMID- 14739487 TI - Practical markers used in the diagnosis of neuroendocrine tumors. AB - Although there are many broad-spectrum neuroendocrine markers, chromogranin and synaptophysin are the principal ones used in diagnostic pathology. Other broad spectrum neuroendocrine markers, transcription factors, and specific peptide markers used in the diagnosis of neuroendocrine tumors are reviewed. The use of different keratins in the differential diagnosis of endocrine tumors is also presented. The importance of using low-molecular-weight keratins such as CAM5.2 to avoid false-negative results in the workup of some neuroendocrine tumors is emphasized. Finally, the use of in situ hybridization in diagnostic pathology is briefly summarized. PMID- 14739486 TI - Oxygen sensing in neuroendocrine cells and other cell types: pheochromocytoma (PC12) cells as an experimental model. AB - A steady supply of oxygen is an absolute requirement for mammalian cells to maintain normal cellular functions. To answer the challenge that oxygen deprivation represents, mammals have evolved specialized cell types that can sense changes in oxygen tension and alter gene expression to enhance oxygen delivery to hypoxic areas. These oxygensensing cells are rare and difficult to study in vivo. As a result, pheochromocytoma (PC12) cells have become a vital in vitro model system for deciphering the molecular events that confer the hypoxia resistant and oxygen-sensing phenotypes. Research over the last few years has revealed that the hypoxia response in PC12 cells involves the interactions of several signal transduction pathways (Ca2+/calmodulin-dependent kinases, Akt, SAPKs, and MAPKs) and transcription factors (HIFs, CREB, and c-fos/junB). This review summarizes the current understanding of the role these signal transduction pathways and transcription factors play in determining the hypoxic response. PMID- 14739488 TI - Paraneoplastic endocrine syndromes: a review. AB - The paraneoplastic endocrine syndromes ("ectopic" or "inappropriate" hormone production) comprise a wide array of symptom complexes associated with malignant or less commonly benign neoplasms. Most of the syndromes are associated with the production of peptide hormones, which, in some instances, have autocrine stimulatory effects. Hypercalcemia, the most common paraneoplastic endocrine syndrome, may be due to the systemic release of parathyroid hormone-related protein (PTHrP), factors that may be produced locally (cytokines), or by a combination of these mechanisms. A spectrum of other syndromes may be related to the production of specific hormones or growth factors, including insulin-like growth factor and fibroblast growth factor 23. Molecular mechanisms responsible for the development of these syndromes are poorly understood. Mutational events not only may initiate neoplastic transformation but may also lead to the activation (re-expression) of genes responsible for hormone production. Additionally, epigenetic events such as methylation may also be responsible for the development of these syndromes. It is likely that a multiplicity of genetic and epigenetic events may contribute to the development of paraneoplastic endocrine syndromes. PMID- 14739489 TI - Potential applications of molecular biology in neuroendocrine tumors. AB - The impact of molecular biology procedures on neuroendocrine (NE) tumor biology is gradually evolving from purely academic and research studies to clinical applications. This review deals with applications of molecular techniques in neuroendocrine tumors, with special reference to their potential for diagnostic, prognostic, or therapeutic impact. Since the cloning of the genes involved in inherited endocrine tumor syndromes, molecular analysis of the responsible genetic alterations has become a routine diagnostic tool to select affected patients and their relatives, and also an interesting approach to investigate the pathogenesis of neuroendocrine tumors. Assessment of the clonal composition of endocrine tumors could be useful to differentiate hyperplastic versus either adenomatous or carcinomatous conditions, as well as to better understand the clonal relationship between different neoplastic populations in mixed tumors. In addition, molecular approaches allow high sensitivity both in defining the neuroendocrine phenotype in poorly differentiated tumors and in searching for micrometastasis during the follow up of patients with endocrine tumors. Finally, the detection of peptide hormone receptors (e.g., oxytocin and somatostatin receptors) and the development of potent synthetic analogs of such peptides, are opening promising applications in the diagnosis and therapy of endocrine tumors. PMID- 14739492 TI - "Honeycomb Golgi" in pituitary adenomas: not a marker of gonadotroph adenomas. AB - The vacuolar change in Golgi complexes known as "honeycomb Golgi" has been described as the ultrastructural hallmark of a specific tumor that has been called the "female gonadotroph" adenoma of the human pituitary. Recently, a few adenomas presenting with Cushing's disease have been reported to exhibit this feature. To clarify the significance of a "honeycomb Golgi" in the classification of pituitary adenomas, we studied clinically nonfunctioning adenomas with or without "honeycomb Golgi" using immunohistochemistry for adenohypophysial hormones and RT-PCR for the cell-specific transcription factors Tpit that identifies corticotrophs and SF-1 that identifies gonadotrophs. All adenomas were from women. Among 20 adenomas with complete "honeycomb Golgi" change, gonadotrophin subunits were totally immunonegative, but ACTH was positive in a few cells of 12 adenomas. Among eight adenomas with partial vacuolar change of the Golgi complex, five were positive for gonadotrophins and two were positive for ACTH. A subgroup of these lesions were examined by RT-PCR and among eight adenomas with typical "honeycomb Golgi" one case expressed both Tpit and SF-1, probably due to contamination with normal pituitary and another expressed neither Tpit nor SF-1. Of the remaining six cases, Tpit was expressed in two cases and SF 1 in four. These findings indicate that "honeycomb Golgi" change can been seen in corticotroph adenomas as well as gonadotroph adenomas. The reason why this vacuolar change occurs only in females remains to be clarified. PMID- 14739491 TI - Ret protein expression in adrenal medullary hyperplasia and pheochromocytoma. AB - Ret is a developmentally regulated tyrosine kinase involved in formation and maintenance of the nervous system. Ret mutations predisposing to pheochromocytomas and medullary thyroid carcinomas occur in multiple endocrine neoplasia (MEN) syndromes 2A and 2B. Biochemical studies have demonstrated overexpression of Ret mRNA and protein in pheochromocytomas compared to normal adrenal medulla. However, the cellular distribution of Ret in the normal human adrenal and in hyperplastic lesions that antecede pheochromocytomas are unclear. The present investigation was undertaken to resolve the histological distribution of Ret in the normal human adrenal, in pheochromocytomas evolving from adrenal medullary hyperplasia in MEN2A and in sporadic pheochromocytomas. Ret expression was studied by immunohistochemistry using both a polyclonal and a monoclonal antibody, with confirmation by immunoblotting of representative cases. Only occasional cells stained for Ret in the normal adrenal, consistent with the distribution in adult adrenals of other species. Heterogeneous, progressively increased Ret expression was observed during the evolution of pheochromocytomas. In both normal and neoplastic adrenal, the most intense immunoreactivity was observed in cells with neuron-like features. Our finding that Ret is not expressed at high levels in the early stages of disease suggests that elucidation of mechanisms that regulate Ret expression is required for understanding the pathobiology of MEN2A. The association of high-level Ret expression with neuronal morphology suggests that the variable overexpression of Ret in pheochromocytomas might in part be an epiphenomenon, reflecting the known phenotypic plasticity of these tumors. PMID- 14739493 TI - Comparison of fine-needle-nonaspiration with fine-needle-aspiration technique in the cytologic studies of thyroid nodules. AB - INTRODUCTION: Fine needle aspiration (FNA) has been adopted as a simple and cost effective technique for cytologic studies of thyroid nodules. The recently introduced fine needle nonaspiration (FNNA) technique, however, is claimed to be superior because it provides specimens with larger number of cells and better preserved cytomorphology and architecture of thyroid papillae and follicles. In this study, we compare the efficacy of the two methods. MATERIALS AND METHODS: Two hundred patients with thyroid nodules, 1-4 cm in size, were recruited. FNA and FNNA techniques were applied to each patient. In a single-blind setting, all specimens were examined by a single cytopathologist. The specimens were scored (0, 1, or 2) on the basis of background blood or clot, number of obtained cells, preserved architecture of papillae and follicles, and cellular degeneration. Nonparametric methods were then used to compare the scores of the two techniques. RESULTS: Two hundred patients (162 female, 38 male) were entered into the study. Specimens from 43 patients were inadequate. In the remaining 157 patients, no statistically significant difference was seen between FNNA and FNA average scores in each parameter (p > 0.05). CONCLUSION: We concluded that FNNA is not superior to FNA in the cytopathologic studies of thyroid nodules. PMID- 14739490 TI - Molecular genetic alterations in adrenal and extra-adrenal pheochromocytomas and paragangliomas. AB - Pheochromocytomas and paragangliomas are neuroendocrine neoplasias of neural crest origin. Genetic mutations that are characterized in other human neoplasms are rarely seen in these tumors. About 10% of the patients with pheochromocytomas and paragangliomas present with a family history of von Hippel-Lindau disease (VHL), Multiple endocrine neoplasia type 2 (MEN2), one of the three familial paraganglioma syndromes (PGL; PGL1, PGL3, PGL4), or neurofibromatosis type 1 (NF1). In an even higher percentage, a genetic predisposition is involved in the development of these tumors. The genes of hereditary tumor syndromes such as the aforementioned ones are also ideal to study the molecular pathogenesis in the sporadic counterparts. Many studies have been undertaken to identify important secondary genetic events that contribute to the tumorigenesis of pheochromocytoma or paraganglioma, but a comprehensive review of these data is lacking. Recent findings of CGH and LOH studies provided new starting points to unravel the pathogenesis and progression of these tumors. This review presents an overview of our current understanding of the molecular pathogenesis of pheochromocytoma and paraganglioma. PMID- 14739495 TI - Sphingosine may have cytotoxic effects via apoptosis on the growth of keloid fibroblasts. AB - Keloids are often resistant to treatment, causing much suffering to the patient. Our previous work found that ceramide (Cer) inhibits growth of fibroblasts via apoptosis. However, when compared to normal fibroblasts (NFs), which are quiescent, keloid fibroblasts (KFs) rapidly proliferate and are reported to be resistant to apoptosis via Cer. Sphingosine (Sph) is a metabolite product of ceramide that has some different biochemical properties. Thereofore, we investigated the cytotoxic effects of Sph on cultured fibroblasts from keloid lesions and normal skin in order to evaluate the possibility of using Sph in the treatment of keloid. We used the lactic dehydrogenase (LDH) method, MTT method, and propidium iodide (PI) method. Sph had cytotoxic effects via apoptosis on both the KFs and NFs. Our results indicate that Sph may be applicable to the future treatment of keloid. PMID- 14739494 TI - Long-term follow up of a "sporadic" unilateral pheochromocytoma revealing multiple endocrine neoplasia MEN2A-2 in an elderly woman. AB - A unilateral, apparently sporadic pheochromocytoma was removed from the right adrenal of a 73-yr-old Caucasian woman. At the time of surgery, germline DNA from the patient was not available. However, a continuous cell line (KNA) established from the tumor showed a heterozygous sequence variant TGC (cysteine) to TGG (tryptophan) in exon 10, codon 611 of the RET proto-oncogene. Subsequent genetic testing of the patient and her offspring revealed the same base-change in herself, one daughter, one son, and the only grandson, confirming hereditary disease classified as MEN2A-2. Clinical follow up of the patient revealed elevated serum calcitonin after 6 yr. Thyroidectomy was performed and revealed a small medullary thyroid carcinoma. The patient's children thus far show no evidence of MEN2, but C-cell hyperplasia has been diagnosed in the grandson. Our serendipitous finding of a MEN2A-2 mutation in a patient with initial diagnosis of late onset, unilateral, "sporadic" pheochromocytoma would argue for routine mutation screening of even elderly patients presenting with a pheochromocytoma. PMID- 14739496 TI - Clinical evaluation of roxithromycin: a double-blind, placebo-controlled and crossover trial in patients with acne vulgaris. AB - We clinically evaluated roxithromycin (ROM) in a double blind, placebo controlled, and crossover trial in patients with inflammatory acne. Patients with inflammatory acne who were attending our outpatient clinic for treatment and who had not received topical or systemic treatment for the previous month were enrolled in this study. Patients were randomly separated into two groups. Group I consisted of 26 patients. The patients received 2x150 mg/day ROM orally in the first period and 2x1 placebo tablets/day in the second period. Group II consisted of 20 patients. These patients received 2x1 placebo tablets/day in the first period and 2x150 mg/day ROM orally in the second period. The first period was the first four weeks, and then there was a washout period of two weeks (5th and 6th). The second period was the next four weeks (7th to 10th weeks) after the washout period. Median acne scores had clearly decreased in both groups at the end of the study. Differences of median acne scores were statistically significant in both groups between at baseline and at the end of the study (p<0.001). The results showed that ROM is a safe and effective alternative in the treatment of inflammatory acne with few side effects and good compliance. PMID- 14739497 TI - A case of anti-epiligrin cicatricial pemphigoid associated with lung carcinoma and severe laryngeal stenosis: review of Japanese cases and evaluation of risk for internal malignancy. AB - A 68-year-old Japanese male with a five-year-history of lung carcinoma showed recurrent blisters and erosions on the oral and genital mucosae and the skin. The patient complained of dyspnea due to severe laryngeal stenosis and underwent a tracheostomy. A skin biopsy specimen showed a subepidermal blister and linear deposits of IgG and C3 at the basement membrane zone of the epidermis. Indirect immunofluorescence examination demonstrated circulating IgG anti-basement membrane zone autoantibodies that reacted to epiligrin on immunoblotting. Based on a diagnosis of anti-epiligrin cicatricial pemphigoid, he was treated with prednisolone, minocycline hydrochloride and nicotinamide. Although no new skin lesions appeared, he died of lung carcinoma five months after the tracheostomy. A review of reported cases with anti-epiligrin cicatricial pemphigoid in Japan disclosed that 5 of 16 cases (31.2%) were complicated by internal malignancies. PMID- 14739498 TI - A case of generalized pustular psoriasis associated with Turner syndrome. AB - We report a 27-year-old Japanese woman with Turner syndrome who had generalized pustular psoriasis of the von Zumbusch type. She developed a febrile diffuse erythema and pustular eruption without any history of preceding psoriasis vulgaris or drug ingestion. Oral treatment with 3.2 mg/kg cyclosporin per day successfully resulted in rapid improvement, followed by a complete remission. To our knowledge, this is the first report describing the unusual coexistence of these two systemic disorders. We discuss a hormone imbalance that might have contributed to the predisposition to pustular psoriasis and difficulties in the management of the patent's treatment. ABBREVIATIONS: TS: Turner syndrome, GPP: generalized pustular psoriasis PMID- 14739500 TI - Benign fibrous histiocytoma of the eyelid with an unusual clinical presentation. AB - Benign fibrous histiocytoma is a common soft tissue tumor that can be deep or superficially located. Although the deep type of fibrous histiocytoma has a predilection for the orbit, the eyelids are an unusual location for the cutaneous type. A 42-year-old woman had bilateral yellowish nodular masses of the eyelids for two years. Pathological examination after excision revealed benign fibrous histiocytoma. Our case is an unusual clinical presentation of cutaneous fibrous histiocytoma as well as a rare location such as the eyelids. PMID- 14739499 TI - Eosinophilic pustular folliculitis following autologous peripheral blood stem cell transplantation. AB - We describe a case of eosinophilic pustular folliculitis (EPF) occurring after autologous peripheral blood stem-cell transplantation for non-Hodgkin's lymphoma and successfully treated with oral and topical indomethacin. Including ours, the reported cases of EPF with HIV-negative hematological disease have shown clinical features similar to those of HIV-associated EPF, however the prognosis is better than that for HIV-associated EPF, which usually shows a persistent course. EPF with HIV-negative hematological disease can be classified as a unique group in EPF. PMID- 14739501 TI - Brooke-Spiegler syndrome associated with cylindroma, trichoepithelioma, spiradenoma, and syringoma. AB - Brooke-Spiegler syndrome is a rare, autosomally dominant disease characterized by the development of multiple cylindromas, trichoepitheliomas, and occasional spiradenomas. We report herein a case of Brooke-Spiegler syndrome in a 55-year old woman associated with multiple cylindromas, spiradenomas, trichoepitheliomas, and syringomas that developed on her face. Her 52-year-old sister also suffered from multiple trichoepitheliomas. Although there is variable clinical and histological expression in Brooke-Spiegler syndrome, all of the tumors encountered in our case were related to folliculosebaceous-apocrine units. PMID- 14739502 TI - Unilateral angiokeratoma scroti: a rare manifestation of a vascular tumor. AB - A 21-year-old patient with a six-year history of asymptomatic papular lesions on his scrotum is reported. The lesions were limited to one half of the scrotum and associated with subclinical varicocele on the same side. The clinical and histopathological findings were consistent with a diagnosis of angiokeratoma scroti. Increased venous pressure caused by the varicocele may have been responsible for the unilateral development of angiokeratoma in this patient. PMID- 14739503 TI - A case of a large dermatofibrosarcoma protuberans successfully treated with radiofrequency ablation and transcatheter arterial embolization. AB - We present a large dermatofibrosarcoma protuberans on a 52 years-old woman's back that was successfully treated with transcatheter arterial embolization and radiofrequency ablation. The tumor developed on her back at the age of 14. Surgical treatment was not complete, and it relapsed at age 17 and again at age 24. The tumor enlarged to over 20x20x10 cm and flooded intermittently. The tumor cells with atypical nuclei were CD34 positive and proliferated in a storiform pattern. These findings were compatible with dermatofibrosarcoma protuberans. She was treated with blood transfusion for severe anemia. Two large feeding arteries were embolized, and the bottom of the tumor was treated with radiofrequency ablation. The tumor reduced in volume by more than 50%. It was then surgically removed and reconstructed with a free mesh skin graft. The combination pretreatment enabled radical resections of a large DFSP without severe complications. PMID- 14739504 TI - Two cases of angiosarcoma of the face. AB - Angiosarcoma is a rare vascular malignant tumor most commonly seen on the scalp of elderly people. We report here two cases of angiosarcoma of the face in 74- and 75-year-old males. It is very unusual to find the development of an angiosarcoma with a rosacea on the face. To the best of our knowledge, only three such cases have been reported. PMID- 14739505 TI - Polypoid Basal cell carcinoma on the perianal region: a case report and review of the literature. AB - We report the case of an 88-year-old Japanese woman with polypoid BCC on the perianal region. The basaloid, solid aggregations of BCC were limited to the upper to middle part of the polypoid area. A review of the English and Japanese literature that excluded cases of fibroepithelioma of Pinkus found 21 total cases in which the polypoid BCC could be recognized as a variant of nodular BCC. The following points were used to categorize the lesions: 1) characteristic polypoid appearance that often mimics clinically benign or malignant adnexal neoplasms, melanocytic nevus, or malignant melanoma; 2) the peculiar favorable locations such as scalp (23.8% of the 21 cases), genital area (23.8% of the cases), and back-buttock areas (19% of the cases). These areas suggest that etiologic factors other than ultraviolet radiation exposure are involved in such cases. These characteristic locations may also be the main reason why fibroepithelioma of Pinkus type BCCs frequently manifest with a polypoid appearance, because these types of BCCs also have similar preferred sites such as the back-buttock and genital areas. Polypoid BCCs appear to affect more women than men than do nodular BCCs (7 men and 14 women). Although the polypoid BCCs have a large average size, they are not considered aggressive because the lesions are well-circumscribed and the growth patterns are non-infiltrative. PMID- 14739506 TI - Delayed diagnosis in a case of lupus vulgaris with unusual localization. AB - Lupus vulgaris is the most common form of cutaneous tuberculosis, and the usual sites of involvement are the head and neck. We present a forty-six-year-old woman with lupus vulgaris on the external surface of the left leg and foot, an unusual site. Based on histopathological and clinical features, this case was diagnosed as lupus vulgaris with unusual localization. PMID- 14739507 TI - Eczema herpeticum localized to area of tinea cruris. AB - Eczema herpeticum (EH) has been reported in association with various skin abnormalities. We experienced a patient with tinea cruris who developed EH within a fungal infection. To our knowledge, dermatophyte infection as an underlying cutaneous disorder for EH has not been previously described in the English literature. PMID- 14739508 TI - Distal phalangeal metastasis of extramammary Paget's disease. AB - A rare case of phalangeal metastasis of extramammary Paget's disease in a 68-year old man is described. The patient developed an erythematous, slightly elevated area in the pubic region. A biopsy specimen demonstrated numerous, large, rounded cells with ample pale-staining cytoplasm proliferating in the epidermis. With a diagnosis of extramammary Paget's disease, he underwent wide local excision and inguinal node dissection. Eleven months postoperatively, the patient developed a tender, red, swollen right ring finger. Bone X-ray showed that the distal phalanx of the ring finger had completely dissolved. Histopathological examination demonstrated proliferation of tumor cells in the adipose tissue. They had poorer and darker cytoplasm than the Paget's cells in the epidermis of the pubic region. Immunohistochemically, these cells showed the same staining pattern as did the Paget's cells at the primary site. Accordingly, the patient was diagnosed with distal phalangeal metastasis of extramammary Paget's disease. Two weeks after the appearance of the distal phalangeal metastasis, the patient died of cancerous pleurisy. It has been reported that patients with phalangeal metastasis have a very poor prognosis. PMID- 14739509 TI - Primary tuberculosis cases presenting with erythema nodosum. AB - Erythema nodosum (EN) is seen only in the primary tuberculosis (TB) form of tuberculous diseases. Among the etiologies of EN, TB is the most frequent disorder in developing countries. We aimed to assess our patients with EN in reference to primary TB. We evaluated 335 patients with the diagnosis of TB during last 20 years; retrospectively 61 (18%) of these cases had pulmonary and 274 (82%) had extrapulmonary TB. Ten (16%) of the pulmonary TB cases were primary. All 10 patients with primary TB presented with EN. Among 50 patients with EN diagnosed and followed during the last 10 years, the etiology was determined in 56%, and primary TB was the most frequent: 20%. PMID- 14739510 TI - Coexistence of psoriasis vulgaris, systemic sclerosis, and annular erythema in association with Sjogren's syndrome. PMID- 14739511 TI - Multiple squamous cell carcinomas arising in lesions of discoid lupus erythematosus. PMID- 14739512 TI - Cutaneous toxicity of chemical irritants on hairless Guinea pigs. AB - To evaluate the toxicity of irritant chemicals on animal skin, investigators have frequently had to apply high concentrations, owing to the fact that its susceptibility is less than that of human skin. High concentrations are so damaging to tissue that specific effects are obscured on the various layers. The aim of the present study was to elucidate the effects of a variety of irritating chemicals on the skin of hairless guinea pigs. Graded concentrations of these irritating substances were applied to the back for varying periods. Histologic changes were analyzed by light and electron microscopy. The structural alterations varied greatly among the chemicals, reflecting quite different mechanisms of action. Hairless guinea pigs are quite susceptible to chemical injury, especially to their hair follicles and dermal components. The hairless guinea pig appears to be an advantageous model to assess the acute and chronic effects of chemical irritants. PMID- 14739513 TI - Roles of stress, stress perception and trait-anxiety in the onset and course of alopecia areata. AB - Clinical observations suggest that the nervous system, including psychological factors, can influence the onset and course of alopecia areata (AA). The aim of this study was to determine whether stressful life events, stress perception, and trait-anxiety are risk factors in the onset and course of AA. A group of 45 patients diagnosed with AA and a group of 45 healthy controls were participants in the study. The patients with AA were divided into two subgroups: patients with a first episode of AA and patients with recidivism of the disease. All participants completed questionnaires addressing sociodemographic, clinical and psychological aspects of their disorder. The frequency and types of stressful life events experienced over the previous six months were recorded. Lemyre and Tessier's Mesure de Stress Psychologique was used to measure emotional, cognitive, behavioral, and physiological aspects of distress. Anxiety was evaluated by the Spielberg's Trait Anxiety Inventory. The subgroups of AA and the control group, using the same numbers of subjects matched for age and sex, education level, marital and employment status, were statistically compared. The number of patients with four stressful life events over the previous 6 months was significantly higher in the group of AA patients with recidivism of disease compared to the control group (P=0.004). There were no differences among the other groups with respect to the frequency of life events. Examination of the relationships between the two groups regarding anxiety, as well as perceived distress, revealed that the groups differed significantly with respect to psychosocial variables studied. A significantly higher degree of trait-anxiety and perceived distress were observed among patients in both AA subgroups (first onset and recidivism of AA) than in the healthy control group. The highest scores for anxiety and stress perception among examined groups were obtained in the group with recidivism of AA (33.42 +/- 12.71 and 90.32 +/- 50.74, respectively). Trait-anxiety and stress perception constitutes risk factors that may influence the onset and exacerbation of AA. The present study does not provide evidence of a significant role of stress in the onset of AA. Life events may play an important role in triggering of some episodes. PMID- 14739514 TI - Inhibition of the proliferation of a malignant peripheral nerve sheath tumor cell line by gamma interferon gene transfection. AB - A solitary malignant peripheral nerve sheath tumor was resected, and a tumor cell line was obtained using an explant culture. The morphology of this cell line was quite similar to those of cell lines of dermal neurofibromas of neurofibromatosis 1, except for the fact that the malignant peripheral nerve sheath tumor cell line could be cultured for more than two months. When the tumor cell line was transfected with human gamma interferon gene, the growth rate was remarkably abolished, as previously observed in neurofibroma cell lines transfected with the gamma interferon gene. This new finding suggests the potential for treating either solitary or neurofibroma-related, inoperable, malignant peripheral nerve sheath tumors by local gamma interferon gene transfection in vivo. PMID- 14739515 TI - Two Brazilian cases of IgA pemphigus. AB - IgA pemphigus is a rare, neutrophilic, acantholytic skin disorder with approximately 70 cases described in the literature. We report two patients with the subcorneal pustular dermatosis (SPD) type of IgA pemphigus. Initially, both patients were misdiagnosed as subcorneal pustular dermatosis of Sneddon and Wilkinson. The correct diagnosis was only made after detecting intercellular IgA depositions in the epidermis by direct immunofluorescence. Immunoblotting (IB) of normal human epidermal extracts, performed on both sera, was negative for Dsg 1, Dsg 3, BP 230, BP 180, 210 kDa envoplakin, and 190 kDa periplakin. ELISA for desmogleins (Dsg 1 and Dsg 3) showed that neither of the cases had IgA antibodies to Dsg. The c-DNA transfection test for desmocollins (Dsc) revealed that the IgA antibodies of both patients reacted with desmocollin 1. This result supports the hypothesis that the autoantigen in SPD type IgA pemphigus is desmocollin 1. PMID- 14739516 TI - Three cases of 'mechanic's hands' associated with interstitial pneumonia: possible involvement with foot lesions. AB - We describe three cases of the rare combination of lateral erythema and hyperkeratosis of the fingers that typify a condition known as 'Mechanic's Hands'. The first and the third cases were unusual in that the condition was associated with cutaneous involvement of the feet and interstitial pneumonia but not with myositis, or with only mild muscular involvement, while the second case was typical, being accompanied by myositis and detectable antibodies against histidyl transferase. We propose that Mechanic's Hands can occur in association with foot lesions and interstitial pneumonia, even if it is not accompanied by myositis. PMID- 14739517 TI - Tinea barbae (tinea sycosis): experience with nine cases. AB - Tinea barbae is a rare dermatophytosis that affects the hair and hair follicles of the beard and mustache. This paper presents 9 cases of tinea barbae observed over an 18-year period of time and classified as follows: 1 was superficial and 8 were deep (6 folliculitis-like and 2 kerion-like). Most of the cases (4) were associated with topical steroid therapy, others with pet contact (3 cases) and one with diabetes. The causal agents isolated were: Trichophyton rubrum in 3; Microsporum canis in 3; Trichophyton mentagrophytes in 2; and Trichophyton tonsurans in one. The involvement of the hair was observed and classified in all cases. The trichophytin skin reaction was positive in all 9 patients. All the patients were treated with systemic antimycotics, 3 cases with griseofulvin, 1 with ketoconazole, 3 with itraconazole, and 2 with terbinafine. Clinical and mycologic cures were achieved at 6 to 8 weeks of treatment at the usual doses. PMID- 14739518 TI - Tinea capitis among children and adolescents in the Farwaniya region of Kuwait. AB - Analysis of 325 patients (182 males, 143 females) of mycologically proven tinea capitis (TC) seen over a period of two years from January 2001--December 2002 in the Farwaniya region of Kuwait is presented in this study. The age range was 8 months to 17 years. Peak incidence was observed in the 3-14 year age group (79.6%). Positive family history and contact with pets were noted in 22% and 36.7% of the cases, respectively. The non-inflammatory 'gray patch' variety was the most common clinical type, seen in 163 (50.2%) children, followed by the black-dot variant in 100 (30.2%) patients. A significant proportion of the cases (16.6%) had the uncommonly reported seborrheic dermatitis or dandruff like pattern. Highly inflammatory kerion was encountered infrequently (2.5%). Seven species of dermatophytes were isolated; Trichophyton violaceum in 135 (41.5%), followed by Microsporum canis in 89 (27.4%), Microsporum audouinii in 48 (14.8%), Trichophyton mentagrophytes var. mentagrophytes in 31 (9.5%), Trichophyton verrucosum in 15 (4.6%), Trichophyton tonsurans in 6 (1.9%) and Microsporum gypseum in 1 (0.3%) patient. T. violaceum was the most common fungus responsible for the black-dot variety (89/100) and kerion (4/8) patients. Three cases of kerion (33.3%) grew T. verrucosum. M. canis was the most common species isolated from the 'gray patch' cases (79/163; 48.5%) followed by almost equal prevalences of T. violaceum (16.6%), M. audouinii (15.3%), and T. mentagrophytes var. mentagrophytes (12.3%). Among the seborrheic type of cases, M. audouinii was the most common fungus isolated in 20/54 (37%) followed by T. violaceum in 15 (27.8%), T. mentagrophytes var. mentagrophytes in 9 (16.7%), M. canis in 8 (14.8%) and T. verrucosum and T. tonsurans in one (0.3%) patient each. The results are significantly different from those in earlier studies in Kuwait. PMID- 14739519 TI - A case of intravascular large B-cell lymphoma with multiple organ involvement. AB - Intravascular large B-cell lymphoma (IVL) is a rare systemic disease characterized by clonal proliferation of neoplastic lymphoid cells within the capillary lumina of small blood vessels. The most common sites of involvement are cerebrovascular or sinusoid areas, resulting in a wide variety of neurologic deficits. Herein we describe a case of IVL that presented with multiple internal and external organ involvements, including the skin, bilateral kidneys, and lung, and with an aggressive clinical course. The confirmative diagnosis was based on the microscopic findings of the skin lesion with a B-cell immunophenotype. The patient suffered from repeated severe dyspnea, hypoxemia, and high-grade fever with a simultaneous increase in serum lactate dehydrogenase and soluble IL-2 receptor levels. These manifestations responded dramatically to intravenous injection of corticosteroid (500 mg/day for 3 days). Despite various clinical presentations, no neurological abnormality was observed until the patient died 12 months after the initial combination chemotherapy. PMID- 14739520 TI - Weight loss-induced calciphylaxis: potential role of matrix metalloproteinases. AB - Calciphylaxis is an uncommon and often devastating syndrome of calcification of small vessels, leading to tissue infarction. The mechanism of how calcium deposits on small vessels is unknown. Recently, metalloproteinase digestion of elastin has been shown to enhance deposition of calcium, suggesting a possible mechanism of calciphylaxis. We describe a case of a patient who developed calciphylaxis after rapid weight loss, but had normocalcemia and normal renal function. She was found to have high levels of matrix metalloproteinases, which may have chemically altered elastin, allowing deposition of calcium on small vessels. Inhibitors of matrix metalloproteinases may be useful in the prevention and treatment of calciphylaxis. PMID- 14739521 TI - P-cadherin expression in skin peeled with phenol or trichloroacetic acid (TCA). AB - P-cadherin expression patterns were studied in trichloroacetic acid (TCA) or phenol treated skin. The expression was absent or very weak on the basal cell surfaces by day 2. Seven days after peeling, P-cadherin was clearly distributed in a continuous granular pattern over the cell surface of the entire epidermis of the 40% TCA treated skin, in a weak granular pattern on a few suprabasal cells and basal cells of the phenol treated skin, and very weakly expressed on the lateral surfaces and in the cytoplasm of basal cells of the 60% TCA treated skin. Based on the present results and previous reports, it is likely that there are distinct patterns of P cadherin expression. Furthermore, a specific type of P cadherin expression might be involved in wound healing in general, which could provide new insights into tissue repair mechanisms after chemical peeling. PMID- 14739522 TI - A case of localized bilateral lipodystrophy associated with self-injection of xenogenous material. PMID- 14739523 TI - A case of embolia cutis medicamentosa. PMID- 14739524 TI - Zosteriform herpetic folliculitis involving eccrine gland. PMID- 14739525 TI - Steroid 5alpha-reductase 1 polymorphisms and testosterone/dihydrotestosterone ratio in male patients with hypospadias. AB - BACKGROUND/AIM: Defects in the steroid 5alpha-reductase type 2 (SRD5A2) activity cause decreased formation of dihydrotestosterone (DHT) from testosterone (T), resulting in defective masculinization of external genitalia; the T/DHT ratio is increased. We investigated 10 patients with elevated T/DHT ratios in whom mutations in the SRD5A2 and AR genes had been excluded to find out whether structural alterations of the SRD5A1 gene could contribute to their genital malformations. METHODS: Single-strand conformation polymorphism analysis and direct sequencing were used to detect variations in the SRD5A1 gene of the patients and of 49 adult fertile men who served as controls. RESULTS: The sequence analysis of exon 3 of the SRD5A1 gene indicated an adenine-to-guanine change (ACA vs. ACG), both triplets encoding the amino acid residue threonine. The ACG sequence was detected in 57% of all subjects and was equally distributed in patients and controls. The T/DHT ratio was significantly higher in controls with the ACG variant as compared with those having the ACA variant. However, no particular sequence aberration was found in the SRD5A1 genes of either group. CONCLUSION: Mutant SRD5A1 isoenzyme does not seem to play a crucial role in the development of hypospadias. PMID- 14739526 TI - Five-year follow-up of a 13-year-old boy with a pituitary adenoma causing gigantism--effect of octreotide therapy. AB - BACKGROUND/AIM: In children, there is little experience with octreotide therapy for pituitary tumors, especially growth hormone (GH) producing adenomas. We report on a 13-year-old boy with gigantism due to a GH-producing pituitary adenoma caused by a Gsalpha mutation on the basis of McCune-Albright syndrome. METHODS: At the age of 6.5 years a GH- and prolactin-producing pituitary adenoma was diagnosed. The adenoma was surgically removed. Immediately thereafter, the small adenoma residuum was treated with octreotide (2 x 100 microg/day s.c.). RESULTS: During therapy with octreotide, the growth rate dropped to normal values; however, rose again after 2 years of treatment. The insulin-like growth factor I (IGF-I) levels remained above the 95th percentile, the GH level mostly >2 microg/l. After 5 years of octreotide therapy, GH (6.9 microg/l), IGF-I (620 microg/l), IGF-binding protein 3 (5.4 mg/l), and prolactin (17.0 ng/ml) levels were still elevated. The growth velocity was +2.4 SDS (standard deviation score), the pubertal status was mature, and the bone age was 14.3 years (prospective final height 208 cm). A magnetic resonance imaging scan showed an unchanged residual 4-mm rim of adenoma at the pituitary site. Side effects from octreotide therapy were not reported by the patient or his family. The therapy was changed to the long-acting release octreotide analog octreotide-LAR. After 1 year of treatment with octreotide-LAR, the GH level was 1.0 microg/l, and the prospective final height dropped by 10 cm. CONCLUSIONS: This case demonstrates that combined surgical and medical treatment can influence the prognosis of childhood gigantism; however, the prognosis of this rare condition remains uncertain. PMID- 14739527 TI - Pituitary metastasis of follicular thyroid carcinoma. AB - OBJECTIVE: The case of a 60-year-old male patient with follicular thyroid cancer who developed a pituitary mass proved to be a metastasis from thyroid cancer. METHODS: Assessment with whole-body scan, ultrasound, computed tomography and thyroglobulin measurements. RESULTS: Despite surgery and repeated doses of radioiodine, the patient developed diplopia and ptosis of the right eyelid, along with increasing thyroglobulin levels. A pituitary mass was discovered, with no signs of pituitary deficiency. The mass was removed and found to consist of neoplastic cells immunohistochemically positive to thyroglobulin. CONCLUSIONS: Distant metastases may develop in cases of follicular thyroid carcinoma, even after repeated doses of (131)I. Metastatic follicular thyroid carcinoma to the pituitary is a rare entity. PMID- 14739528 TI - The link between exposure to dioxin and endometriosis: a critical reappraisal of primate data. AB - Endometriosis is a common and enigmatic disease affecting women of reproductive age. In 1993, Dr. Sherry Rier and her colleagues reported a serendipitous finding that quickly sent a shock wave through the endometriosis research community. They found that rhesus monkeys exposed daily for 4 years to dioxin developed endometriosis, with incidence and severity related to dose. The study prompted more animal and epidemiologic studies regarding the link between dioxin exposure and endometriosis. Yet, 10 years after the first piece of evidence was reported, the primate data are still equivocal, and the human data supporting the dioxin endometriosis association are scanty and conflicting. While many reviewers of the subject recognize the need for more data, other reviewers tend to discount negative studies when reviewing positive studies. In this paper, a critical reappraisal of all evidence from human and primate data is presented. While there is evidence suggesting that exposure to dioxin may facilitate the short-term survival of endometrial implants in non-human primates, this evidence is not supported by both human and non-human primate studies evaluating the relationship between dioxin exposure and the development of spontaneous endometriosis. Weighing all converging evidence, it seems that there are no solid, credible data available at this moment to support the hypothesis that dioxin exposure may lead to the development of endometriosis. PMID- 14739529 TI - Paradigms for mechanical signal transduction in the intestinal epithelium. Category: molecular, cell, and developmental biology. AB - Diverse physical forces including deformation or strain, pressure, and shear stress affect the intestinal mucosa during normal function, and mucosal biology is altered in pathological states in which these forces alter. Taken together with evidence in other tissues and cell types that physical forces can affect cell biology, this has led to the hypothesis that repetitively applied physical forces can initiate intracellular signals that alter intestinal epithelial proliferation and phenotype. This review outlines the nature of such forces and summarizes in vivo and in vitro evidence in support of the paradigm that repetitive force is trophic for the intestinal mucosa via a complex cascade of intracellular signals. PMID- 14739530 TI - Patients with Alzheimer's disease may be particularly susceptible to adverse effects of statins. AB - In epidemiological, cross-sectional studies, treatment with 3-hydroxy-3 methylglutaryl coenzyme A reductase inhibitors (statins) prevented to a large extent the development of Alzheimer's disease (AD), but the results of randomized, placebo-controlled studies, focused on statin therapy in patients with ischemic heart disease (IHD), are at variance. Nonetheless, data from epidemiological, longitudinal studies in humans as well as studies on transgenic mouse models and cultured neuronal cell lines indicate that cholesterol may contribute to the pathogenesis of AD. Statins have proven therapeutic and preventive effects in IHD and other vascular diseases in man. They generally are well tolerated, but some adverse effects, probably due to antiproliferative and proapoptotic properties of the statins, are matters of concern. AD patients may be extrasusceptible to adverse effects of statins due to preexisting aberrations in signal transduction and energy metabolism in the neurons and a perturbed cholesterol metabolism in the brain. This problem might be addressed in randomized, double-blind studies with statins in AD. The statins differ from each other in several aspects, and they are not considered to be therapeutically interchangeable. It could be fruitful to use both a placebo and two different types of statins, i.e. an essentially hydrophilic statin and a lipophilic statin, in a double-blinded fashion, and to compare the effects on the cognitive decline in AD. PMID- 14739531 TI - Efficacy of fluvoxamine as a treatment for behavioral symptoms in frontotemporal lobar degeneration patients. AB - Patients with frontotemporal lobar degeneration (FTLD) present a profound personality change, social misconduct, overeating, and stereotyped behavior. We examined the hypothesis that many of the behavioral symptoms of FTLD will respond to selective serotonin reuptake inhibitors (SSRIs). Sixteen FTLD patients were treated with an SSRI (fluvoxamine maleate) in an open 12-week trial. Treatment responses for stereotyped behavior and other neurobehavioral symptoms were evaluated by the Stereotypy Rating Inventory and the Neuropsychiatric Inventory. The behavioral symptoms, especially stereotyped behaviors of FTLD, significantly improved after treatment. Randomized, placebo- and other SSRI-controlled trials may improve available treatments. PMID- 14739532 TI - Regional analysis of differently phosphorylated tau proteins in brains from patients with Alzheimer's disease. AB - Neurofibrillary tangles (NFT) in Alzheimer's disease (AD) are composed of abnormally phosphorylated tau proteins. Many phosphorylation sites have been reported in the AD brain, and NFT distribution was now roughly classified into 3 stages by Braak stage; this classification is based on pathological studies using the specific silver impregnation technique. The aim of our study was to examine the regional distribution of differently phosphorylated tau proteins with 5 site specific monoclonal antibodies against the tau proteins, AT8, AT180, HT7, Tau2 and Tau5. We then compared our findings with those obtained from silver-stained NFT in an attempt to clarify the relationship between abnormal phosphorylation sites of the tau protein and NFT development. AT180 and AT8 labeled the highest and Tau2 the lowest density of NFT in any regions, while Tau5 and HT7 showed inconsistent distribution. In the limbic cortex, cornu ammonis, entorhinal cortex and cingulate cortex, silver-stained NFT density significantly correlated with density of NFT labeled with the 5 anti-tau antibodies, but cerebral isocortices showed heterogenous patterns of tau-positive NFT. Quantification of tau-positive regional NFT density showed that the AD-associated phosphorylation process progresses from the C-terminal to the N-terminal of the amino acid sequence, and correlation of Gallyas-stained NFT density with tau-labeled NFT density was more significant in the limbic cortices than the cerebral isocortices, which implies that stereotypical phosphorylation occurs in the limbic structures. PMID- 14739533 TI - Apolipoprotein E gene polymorphism in Indian patients with Alzheimer's disease and vascular dementia. AB - The association of apolipoprotein E (ApoE) gene polymorphism with Alzheimer's disease (AD) has been reported in several populations including one from a rural community in North India. However, the association of ApoE polymorphism with vascular dementia (VaD) is yet to be established in this population. In a case control study involving 54 cases of dementia (29 AD and 25 VaD) and 76 age matched healthy controls, the frequency of epsilon4 allele was significantly higher among cases of AD and VaD compared with controls (p < 0.001). The epsilon3epsilon3 (p < 0.05) and epsilon2epsilon3 (p < 0.001) genotypes were found to be protective. The odds of developing AD or VaD were 4.4 and 3.7 times higher, respectively, in the presence of even a single epsilon4 allele. Our results suggest that the increased risk of developing AD or VaD is similar among Asian Indians with ApoE epsilon4 compared with the Caucasian population. PMID- 14739534 TI - The course of cognitive decline in older persons: results from the longitudinal aging study amsterdam. AB - The course of cognitive functioning in older persons was studied over a period of 6 years. The first aim was to distinguish cognitive decline as a temporary state from progressive cognitive decline. The second aim was to identify predictors which may be useful in discriminating persons with temporary cognitive decline from persons with progressive cognitive decline at an early stage. Data were derived from the Longitudinal Aging Study Amsterdam (LASA), a population-based study in the Netherlands. The results show that 18.2% of the sample of older persons showed cognitive decline over a period of 3 years. Of this group, 44% recovered from cognitive decline or stayed stable in the next 3 years. Especially older age, memory complaints and an increase of cardiovascular diseases at follow up predict further deterioration. PMID- 14739535 TI - The Fas promoter polymorphism at position -670 is not associated with late-onset sporadic Alzheimer's disease. AB - The Fas antigen is a cell surface receptor-mediating cell apoptosis. Recent studies have demonstrated that Fas-associated apoptosis is involved in the pathogenesis of Alzheimer's disease (AD). Moreover, the Fas gene is located on chromosome 10q24.1, a region of linkage to late-onset AD (LOAD). These two criteria, pathobiological and positional, make the Fas antigen an interesting candidate for an association with AD. We performed a case-control association study between the common A/G polymorphism at position -670 in the Fas gene (TNFSRF6) promoter and sporadic AD in Jews, investigating whether this locus acts as a risk factor or whether it has a modifying effect. An association has recently been detected by Feuk et al. in the Scottish population between this locus and the risk of early-onset AD (EOAD), but not of LOAD. In agreement with Feuk et al., we found no association between this locus and the risk of LOAD (n = 86). However, in our small sample of patients with EOAD (n = 19), no association was found either. No interactive effect was found between the Fas promoter polymorphism at position -670 and the known risk factor of LOAD, apolipoprotein E epsilon4, and no association was detected with disease progression. These findings show no evidence for an association between the Fas promoter polymorphism at position -670 and AD in our population. PMID- 14739536 TI - The regulatory region polymorphisms of the MTHFR gene are not associated with Alzheimer's disease. AB - Recent epidemiological studies have emphasized the impact of elevated blood homocysteine levels on the pathogenesis of Alzheimer's disease (AD). In spite of a significant impact of a MTHFR C677T polymorphism on the blood homocysteine levels, the association between the C677T polymorphism and AD remains controversial. Therefore, other unidentified genetic factor(s) that regulate blood homocysteine levels may exist. Here, we have analyzed the 5'-upstream region of the MTHFR gene and examined AD patients (n = 223) and nondemented individuals (n = 323) for polymorphisms in the 5'-upstream region of the MTHFR gene. We identified two polymorphisms (-713G/A and -393C/A, upstream of the start codon). We found no significant relationship between AD and the 5'-upstream region polymorphisms of the MTHFR gene. Thus, our study does not reinforce the hypothesis of an independent involvement of the MTHFR gene upstream region polymorphisms in AD risk. PMID- 14739537 TI - Naringenin from Citrus junos has an inhibitory effect on acetylcholinesterase and a mitigating effect on amnesia. AB - This study was performed to identify safe and more effective acetylcholinesterase (AChE) inhibitors in the treatment of Alzheimer's disease. The total methanol extract of Citrus junos had a significant inhibitory effect on AChE in vitro. By sequential fractionation of C.junos, the active component was finally identified as naringenin. Naringenin inhibited AChE activity in a dose-dependent manner. In this study, we also evaluated the anti-amnesic activity of naringenin, a major flavanone constituent isolated from C. junos, in vivo using ICR mice with amnesia induced by scopolamine (1 mg/kg body weight). Naringenin, when administered to mice at 4.5 mg/kg body weight, significantly ameliorated scopolamine-induced amnesia as measured in both the passive avoidance and the Y-maze test. These results suggest that naringenin may be a useful chemopreventive agent against Alzheimer's disease. PMID- 14739538 TI - Results of quinacrine administration to patients with Creutzfeldt-Jakob disease. AB - Several chemicals inhibit the accumulation of abnormal prion proteins in vitro. We administered one, the antimalarial agent quinacrine, to three patients with sporadic Creutzfeldt-Jakob disease (CJD) and to one with iatrogenic CJD. Quinacrine at 300 mg/day was given enterally for 3 months. Within 2 weeks of administration, the arousal level of the patient with akinetic mutism improved. The other 3 patients, insensible before treatment, had integrative responses such as eye contact or voluntary movement in response to verbal and/or visual stimuli restored. Clinical improvement was transient, lasting 1-2 months during treatment. Quinacrine was well tolerated, except for liver dysfunction and yellowish pigmentation. Although its antiprion activity in the human brain has yet to be proved, these modest effects of quinacrine suggest the possibility of using chemical intervention against prion diseases. PMID- 14739539 TI - Association of the Neprilysin gene with susceptibility to late-onset Alzheimer's disease. AB - Neprilysin (NEP), also known as neutral endopeptidase, enkephalinase, CD 10, and common acute lymphoblastic leukemia antigen, is a 97-kD protein. NEP can degrade amyloid beta peptides, and its mRNA and protein levels are known to be reduced in the brains of patients with Alzheimer's disease (AD), making the NEP gene a substantial candidate for an AD risk factor. We examined the genetic association of three NEP polymorphisms, a GT-repeat polymorphism and two single nucleotide polymorphisms (SNPs, -1075A>G and -1284G>C) in its promoter region, with AD in a Japanese case-control sample (240 patients and 163 controls). The GT-repeat polymorphism, but not the SNPs, was significantly associated with late-onset AD (p = 0.0007). Our findings suggest that the GT-repeat polymorphism in the promoter region of the NEP gene or some other unknown polymorphisms, which are in a linkage disequilibrium, confer a susceptibility to late-onset AD. PMID- 14739540 TI - An interleukin-6 promoter variant is not associated with an increased risk for Alzheimer's disease. AB - Recent studies have implicated interleukin-6 (IL-6) in the pathogenesis of Alzheimer's disease (AD). Neuro-inflammatory processes surrounding the amyloid plaques contribute to the progression of AD-related neurodegeneration. IL-6 is a multifunctional inflammatory cytokine which possibly acts as a mediator in the local immune response in the brain of AD patients. In this study we investigated whether the risk of developing AD is altered in carriers of the C allele of a G/C polymorphism at position -174. 113 AD patients and 108 age- and gender-matched nondemented control subjects were analysed. Genotyping of IL-6 was performed using standard PCR and restriction fragment length polymorphism methods. The results were adjusted for age, gender and apolipoprotein E epsilon4 status. There was no evidence for an association between the polymorphism and the risk of developing AD. No evidence of an earlier age at onset for carriers of the C allele was evaluated. We conclude that IL-6 (-174) polymorphism does not influence the risk of developing AD in our cohort. PMID- 14739541 TI - Getting lost: directed attention and executive functions in early Alzheimer's disease patients. AB - This study explores the link between directed attention (DA) and getting lost behavior (GLB) in early Alzheimer's disease (AD) using a cross-sectional design with 3 groups. Based on their dementia levels, 116 community-dwelling participants were recruited from a teaching hospital in Taiwan and classified as the non-demented control, questionably demented, and mild AD groups. Statistical analyses include Pearson correlations, one-way ANOVA, and multiple regressions. Attentional impairments, consisting of distractibility, impulsivity, and executive function problems, significantly predict GLB in familiar and unfamiliar environments. Irritability and executive function problems are associated with mental difficulties in choosing a turn, whereas the use of way-finding strategies reduces GLB. Future interventions may include: (a) mental hygiene of aging; (b) programs targeted at improving attentional function and effective way-finding, and (c) inclusion of DA tests in a routine clinical neuropsychological examination for early detection and accurate diagnosis of dementia. PMID- 14739542 TI - Awareness of deficits in mild cognitive impairment and Alzheimer's disease: do MCI patients have impaired insight? AB - In this study we investigated impaired awareness of cognitive deficits in patients with mild cognitive impairment (MCI) and Alzheimer's disease (AD). Very few studies have addressed this topic, and methodological inconsistencies make the comparison of previous studies difficult. From a prospective research program 36 consecutive patients with mild AD (MMSE above 19), 30 with amnesic MCI and 33 matched controls were examined. Using three methods for awareness assessment we found no significant differences in the level of awareness between MCI and AD. Both groups had impaired awareness and significant heterogeneity in the clinical presentation of awareness. The results demonstrate that subjective memory problems should not be a mandatory prerequisite in suspected dementia or MCI, which makes reports from informants together with thorough clinical interview and observation central when assessing suspected dementia disorders. PMID- 14739543 TI - Semantic knowledge of famous people in mild cognitive impairment and progression to Alzheimer's disease. AB - Patients with dementia of Alzheimer's type (DAT) show severe impairment in recognizing famous people. The aim of the current study was to investigate if this well-known memory impairment of famous faces is already present in the preclinical phase of DAT and if the famous faces test can help to differentiate patients with mild cognitive impairment (MCI) who progress to dementia and those who do not. We compared baseline performance in a task of famous face identification in a sample of 116 patients with subjective memory complaints classified in three groups: 17 participants with no evidence of cognitive impairment; 26 patients with MCI who had not developed dementia, and 27 patients with MCI who had developed probable DAT 2 years later. The remaining patients were excluded because they abandoned or did not meet the applied restrictive criteria for DAT, MCI or control. MCI patients who were diagnosed 2 years later with DAT performed significantly worse in the preclinical phase than MCI and control participants (p < 0.004). Patients with MCI but not DAT obtained intermediate results between control subjects and MCI patients who develop Alzheimer's disease. A neuropsychological task of semantic knowledge of famous people may be useful in the early diagnosis of Alzheimer's disease. PMID- 14739544 TI - Incidence and risk factors for mild cognitive impairment: a population-based three-year follow-up study of cognitively healthy elderly subjects. AB - BACKGROUND: Mild cognitive impairment (MCI) has attracted considerable interest as a potential predictor of Alzheimer's disease (AD). Both the apolipoprotein E (ApoE) epsilon4 allele and vascular factors have been associated with a higher risk for AD, recently they have also been linked to the risk of MCI. OBJECTIVES: To estimate the incidence of MCI among cognitively healthy elderly subjects during a 3-year follow-up, and to evaluate the impact of demographic and vascular factors as well as the ApoE epsilon4 allele on the conversion to MCI. METHODS: At baseline, the cognitive abilities of 806 out of 1,150 eligible subjects (aged 60 76 years) from a population-based sample were examined. Cognitively intact subjects (n = 747) were followed for an average of 3 years. RESULTS: 66 subjects (8.8%) had converted to MCI. The global incidence rate of MCI was 25.94/1,000 person-years. Persons with higher age (OR 1.08, 95% CI 1.01-1.16), ApoE epsilon4 allele carriers (OR 2.04, 95% CI 1.15-3.64) and persons with medicated hypertension (OR 1.86, 95% CI 1.05-3.29) were more likely to convert to MCI than those individuals of lower age and without an ApoE epsilon4 allele or medicated hypertension. Persons with high education (OR 0.79, 95% CI 0.70-0.89) were less likely to convert to MCI than persons with low or no education. In subjects with both the ApoE epsilon4 allele and medicated hypertension, the crude OR for conversion was 3.92 (95% CI 1.81-8.49). In subjects with cardiovascular disease, the crude OR for conversion was 2.13 (95% CI 1.26-3.60). Gender, elevated blood pressure, diabetes or cerebrovascular disease had no significant effect on the conversion to MCI. CONCLUSION: Higher age, the presence of at least one ApoE epsilon4 allele and medicated hypertension are independent risk factors, but high education is a protective factor for MCI. The results suggest that vascular factors may have an important role in the pathogenesis of MCI. PMID- 14739545 TI - CSF lactate dehydrogenase activity in patients with Creutzfeldt-Jakob disease exceeds that in other dementias. AB - The diagnosis of Creutzfeldt-Jakob disease (CJD) is still made by exclusion of other dementias. We now evaluated lactate dehydrogenase (LDH) in the cerebrospinal fluid (CSF) as a possible additional diagnostic tool. CSF LDH levels of patients with CJD (n = 26) were compared with those in other dementias (n = 28). LDH isoenzymes were determined in a subset (n = 9). Total LDH and isoenzyme LDH-1 were significantly higher, whereas the fractions of LDH-2 and LDH 3 were significantly lower in CJD patients. We conclude that in addition to established CSF parameters, LDH and its isoenzymes might serve as a further help to discriminate between CJD and other dementias. PMID- 14739546 TI - Regional cerebral blood flow in Alzheimer's disease: classification and analysis of heterogeneity. AB - Neural networks have been successfully applied to brain perfusion images to classify patients with Alzheimer's disease from normal or other patient populations. Given the recognition that Alzheimer's disease constitutes a heterogeneous disorder, the identification of subgroups sharing common functional brain deficits would constitute a further improvement in the utility of such methods. Therefore, we aimed to investigate whether neural networks could discriminate cortical perfusion deficits of patients with Alzheimer's disease from normal brain perfusion. A second step was to identify subgroups of patients sharing similar perfusion deficits. The study population consisted of one group of 92 normal healthy subjects and one group of 132 patients with mild-to-moderate Alzheimer's disease. The patients were diagnosed according to established criteria (DSM-IV and NINCDS-ADRDA). Regional cerebral blood flow was assessed by the non-invasive (133)Xe inhalation method, using a 64-detector system for measurements of blood flow in superficial cortical areas. The regional blood flow values were used as the only input to artificial neural networks with multilayer Perceptron architecture. The networks were trained using the back-propagation updating algorithm. A fourfold cross validation procedure was used in order to obtain the most reliable performance of the networks. The performance of the neural network, measured as the area under the receiver-operating characteristic curve, was 0.94, with a sensitivity for Alzheimer's disease of 86% at a specificity of 90%. An analysis of the relative importance of cortical areas in the discrimination showed that left parietal areas were more important than the right homologous ones. A clustering analysis of the Alzheimer patients identified three or four subgroups of patients with clearly different combinations of blood flow pathology. A consistent finding in all subgroups was a significant deficit in temporoparietal blood flow of both hemispheres. Distinct group differences were seen in frontal, central and occipital areas with different combinations of involvement. This is the first study in which neural networks have been applied to brain perfusion images obtained with the (133)Xe inhalation method. The results demonstrate that a classification of patients with Alzheimer's disease obtained with this method is compatible with the best results obtained with other brain imaging methods. The identification of clearly distinguishable patterns of blood flow pathology in subgroups of patients lends further support to the notion that Alzheimer's disease is a heterogeneous disorder. PMID- 14739547 TI - Transcobalamin polymorphism and serum holo-transcobalamin in relation to Alzheimer's disease. AB - Isoforms of the vitamin B(12) carrier protein transcobalamin (TC) might influence its cellular availability and contribute to the association between disrupted single-carbon metabolism and Alzheimer's disease (AD). We therefore investigated the relationships between the TC 776C>G (Pro259Arg) genetic polymorphism, total serum cobalamin and holo-TC levels, and disease onset in 70 patients with clinically diagnosed AD and 74 healthy elderly controls. TC 776C>G polymorphism was also determined for 94 histopathologically confirmed AD patients and 107 controls. Serum holo-TC levels were significantly higher in TC 776C homozygotes (p = 0.04). Kaplan-Meier survival functions differed between homozygous genotypes (Cox's F-Test F(42, 46) = 2.1; p = 0.008) and between 776C homozygotes and heterozygotes (Cox's F test F(46, 108) = 1.7; p = 0.02). Proportionately fewer TC 776C homozygotes appear to develop AD at any given age, but this will require confirmation in a longitudinal study. PMID- 14739548 TI - The fuzzy frontier between subjective memory complaints and early dementia. A survey of patient management in German memory clinics. AB - BACKGROUND: Mild cognitive decline is frequent in the elderly population. Whether it is related to normal aging or an early phase of evolving dementia is difficult to ascertain with confidence, and accordingly there is a lack of consensus guidelines for diagnosis and therapy in such patients. We assessed the variability with which memory clinics deal with this problem in everyday practice. METHODS: We sent three fictitious case histories to all 85 German memory clinics that contained the results of clinical examination and neuropsychological test scores and asked for diagnosis and patient management. Patient 1 presented with complaints of mental decline but normal neuropsychological and neurological evaluation and normal daily living activities. Patient 2 came in as a control subject for a study and had impaired test scores but preserved daily living activities, and patient 3 was brought in by relatives with slight impairment of daily living activities and decline in some neuropsychological test scores but relatively spared memory scores. RESULTS: Most of the 51 respondents agreed in recommending further neuropsychological testing, a basic laboratory work-up, brain imaging, and a re-examination after 3 6 months. Yet, there was a high variability in the diagnostic terms used, in the additional diagnostic procedures proposed, and in the recommendations concerning therapeutic intervention and driving. CONCLUSIONS: The results reveal a need of practice guidelines for the use of diagnostic terms, therapeutic interventions and driving recommendations in patients between subjective memory complaints and early dementia. PMID- 14739549 TI - The interrelatedness of wandering and wayfinding in a community sample of persons with dementia. AB - The aim of this study was to evaluate the relationship of wandering and wayfinding and validate the Revised Algase Wandering Scale - Community Version (RAWS-CV) using a community sample of persons with dementia. Adult caregivers (n = 266) completed the RAWS-CV and the Wayfinding Effectiveness Scale (WES). Four aspects of wandering were confirmed (persistent walking, repetitive walking, spatial disorientation, eloping behavior), and two new aspects were also validated (negative outcomes, mealtime impulsivity). The spatial disorientation subscale of the RAWS-CV had significant (p < 0.01) negative correlations with all WES subscales. The global strategies and simple wayfinding goals subscales of the WES correlated significantly with all RAWS-CV subscales except repetitive walking and mealtime impulsivity. ANOVAs comparing wayfinding at 4 levels of wandering revealed differences only for the simple wayfinding goals subscale. Studies examining the relationship of wandering and wayfinding at various levels of cognitive impairment are suggested to further understand these phenomena. PMID- 14739550 TI - Activation of the inflammatory reaction within minutes after birth in ventilated preterm lambs with neonatal respiratory distress syndrome. AB - To study the activation of the inflammatory reaction within minutes after birth, we measured parameters of inflammation before and immediately after birth. To assess whether respiratory distress syndrome (RDS) or birth itself initiates activation, we compared preterm ventilated lambs with term nonventilated lambs. Preterm lambs were delivered by cesarean section at 132 days gestational age (term 145 days) and were ventilated by conventional ventilation (n = 9). Before clamping the cord, 5, 10 and 15 min after birth, blood was sampled from umbilical catheters. Term lambs (n = 9) were born spontaneously after 140-145 days gestational age. Immediately after birth, a venous umbilical catheter was inserted. Blood was sampled before the first breath and 5, 10, 15 and 20 min after birth while the lamb was breathing spontaneously. Blood was analyzed for AP50 (complement activation), number of polymorphonuclear leukocytes (PMNs) and beta-glucuronidase (released from activated PMNs). In preterm lambs, we found a decreased number of PMNs and increased levels of beta-glucuronidase already at 5 min after birth. In the term lambs, we found only a short-term mild decrease in PMNs and short-term increase in beta-glucuronidase. We conclude that systemic activation of the inflammatory reaction can be found in ventilated preterm lambs with RDS within 5 min after birth. This very early activation is mild, transient and less pronounced in term-born spontaneously breathing lambs compared with preterm, ventilated lambs with RDS. PMID- 14739551 TI - Correlation between plasma and urine phenylalanine concentrations. AB - In this pilot study, we show that plasma phenylalanine concentration can be predicted from urine concentration if the age of the patient is taken into consideration. This observation could open the way to a new monitoring of phenylketonuric patients in which painful frequent blood sampling, mandatory to adapt the low phenylalanine diet, could be mostly replaced by urinalysis. Compliance to treatment would be improved and hence also the ultimate mental development. Since this study was based on a small number of patients, validation of the model in a large multicentric survey is needed before it can be recommended. PMID- 14739552 TI - Hematological parameters and cobalamin status in infants born to smoking mothers. AB - Hematological parameters, serum cobalamin and folate levels, and the concentrations of the functional markers plasma methylmalonic acid and total homocysteine were determined in 173 newborns and 46 infants at 6 weeks to see whether maternal smoking influences the hematological parameters and the vitamin status of the newborn. At birth, there was a strong inverse correlation between the number of cigarettes smoked per day during pregnancy and red blood cell count (r = -0.56, p = 0.001) and hemoglobin level (r = -0.52, p = 0.003) in the newborns. Neonates born to smoking mothers had lower red blood cell counts and lower hemoglobin and serum cobalamin levels as compared with infants born to nonsmoking mothers. At 6 weeks, maternal smoking significantly predicted the methylmalonic acid and total homocysteine levels, suggesting an influence from smoking on the cobalamin function in these infants. PMID- 14739553 TI - Gender determines long-lasting effects on adult offspring heart after early-life malnourishment. AB - This study evaluates the effect of insulin in the association between nutrition and gender during lactation on inducing changes in heart morphology of pups when adults. Two groups of lactating rat dams were fed a normal diet containing 22% protein or 0% protein during the first 10 days of lactation. Eight-day-old rats from the female control group (FC) displayed a heart weight/body weight (H/B) ratio 100% greater when compared with the male control group (MC). An increased H/B ratio (65%) was found in the MC group when compared with the male malnourished group (MM). An effect on the development was present in the FC group in which, at 8 days, the H/B ratio was 70% greater than in this group at 60 days of age. A similar effect was observed in the female malnourished group (FM), where the H/B ratio was 100% greater. The effect on the development, at 8 days, in the FC group, when compared with the females at 60 days, displayed a larger cardiomyocyte volume [QA(c)] and a higher cardiomyocyte density [Vv(c)]: QA[c] 220% greater (p < 0.05) and Vv[c] 530% greater (p < 0.01). The FM group, at 8 days of age, had the QA[c] and Vv[c] greater than at the age of 60 days: QA[c] 100% greater (p < 0.05) and Vv[c] 290% greater (p < 0.05). The insulin/glucose ratio reflected the variable gender associated with the malnourishment in the male animals at 8 days of age. In these animals the insulin/glucose ratio of the malnourished animals was found to be reduced by 42% when compared with the control groups. At 60 days of age, a difference in the insulin/glucose ratio was present between FC and FM groups (33% greater in the FC group, p < 0.05). Gender was associated with a high insulin/glucose ratio in MC and MM groups when compared with the FC and FM groups. Also this association was observed in the animals at 60 days of age when the insulin/glucose ratio of the MM animals was 45% greater than that observed in the FM group. PMID- 14739554 TI - Ciprofloxacin treatment in newborns with multi-drug-resistant nosocomial Pseudomonas infections. AB - OBJECTIVES: To evaluate the efficacy and acute side effects of ciprofloxacin treatment in newborns who developed nosocomial Pseudomonas aeruginosa infection. METHODS: Intravenous ciprofloxacin treatment was given to 30 newborns who developed nosocomial P. aeruginosa infection as proven by culture antibiogram results. Initial doses of 10 mg/kg/day were given and increased up to 40 mg/kg/day according to clinical response, laboratory and culture results. During therapy, complete white blood cell counts, urinalysis, liver and renal function tests were performed weekly. All patients were examined daily during treatment for possible symptoms of joint toxicity such as erythema and swelling. The patients were evaluated by general physical examination, with special attention to joints, 1 week after discharge. RESULTS: Two of the patients (6.6%) died due to pseudomonas infection, but the bacteria were successfully eradicated in 28 patients (93.4%). Four patients died from other causes. No laboratory abnormality related to ciprofloxacin was observed during treatment. Swelling and hyperemia of the joints were not encountered during treatment and the 1-week period after discharge. Ciprofloxacin-resistant P. aeruginosa isolates were not grown during the study. CONCLUSION: Ciprofloxacin treatment is effective in life-threatening multi-drug-resistant P. aeruginosa infections. PMID- 14739555 TI - Cord blood cardiac troponin T and troponin I levels in multiple-gestation neonates. AB - The increased mortality and morbidity rates in multiple-gestation neonates are not completely understood. Troponin measurements have a role in situations where the evaluation of the cardiac damage is difficult, such as in cases of unexplained intrauterine fetal growth restriction or death. These conditions, along with perinatal hypoxic risk and in utero ischemic damage, are frequently found in multiple gestations. In this context, a myocardial damage could be expected more frequently in multiple than in singleton births. We hypothesized that cord blood cardiac troponin T and troponin I, markers of myocardial damage, could be different between singleton and multiple pregnancies and, among twins, between the first- and the second-born twin. Troponins T and I and creatine kinase MB concentrations were not increased in twins at birth and were not different between the first- and the second-born twin. These data suggest that myocardial damage, evaluated by cardiac troponin T, troponin I, and creatine kinase MB measurements, does not seem to be a relevant problem in multiple gestation neonates. PMID- 14739556 TI - Lung microvascular adaptation in infants with chronic lung disease. AB - Microvascular development is critical for normal lung maturation. The aims of this study were (1) to quantitatively and qualitatively assess lung microvascular growth in the human fetus, from 22 to 40 weeks' gestation, and (2) to compare development in these infants to those with mild, moderate and severe chronic lung disease (CLD). Using 1- and 4-microm thick sections and electron microscopy, lungs were morphometrically assessed for surface density of distal air spaces; volume density of parenchymal vessels having an air-blood barrier (ABB); percent of distal air space wall having an ABB, and capillary loading, defined as ABB/mm2 of epithelial surface area. The percent of vessels with ABB increased in controls during development in parallel with increasing lung parenchyma. Infants with severe CLD had fewer ABBs and less capillary loading than controls up to 34 weeks' post-conceptional age (PCA), but by 36-40 weeks, showed catch-up growth. Microvasculature vessel diameter, septal thickness, and air sac diameter at 36-40 weeks' PCA were increased with severe CLD, and vessels were more distant from the air surface. We conclude that infants with severe CLD have both stunted secondary septation and microvascular development, but over time, the primary septal wall adapts by thinning and increasing the number of ABBs, thereby taking on the function of secondary septa. PMID- 14739557 TI - Circadian disorganization in experimental arthritis. AB - This review discusses the experimental evidence indicating that arthritis disrupts circadian organization, which was mainly derived from animal studies employing Freund's complete mycobacterial adjuvant (FCA). The defense response to antigenic challenge, mediated in part by cytokines, includes changes in chronobiological central nervous system function, like depressed daily activity, superficial sleep or anorexia. Interferon (IFN)-gamma receptors are detectable in the central circadian pacemaker, the hypothalamic suprachiasmatic nuclei, at a time when the capacity for photic entrainment of the pacemaker became established. The disruptive effects of the systemic injection of IFN on the circadian rhythms of locomotor activity, body temperature and clock-gene mRNA expression have been documented. In the last few years we have examined a number of immune and neuroendocrine circadian rhythms in FCA-injected rats, both in the preclinical phase of arthritis (2-3 days after FCA injection) as well as in the acute phase of the disease (18 days after FCA injection). In arthritic rats, the 24-hour organization of immune and neuroendocrine responses becomes altered. A hormonal pathway involving the circadian secretion of melatonin and a purely neural pathway including, as a motor leg, the autonomic nervous system innervating the lymph nodes were identified. The significant effects of the immune-mediated inflammatory response on the diurnal rhythmicity of adenohypophysial and hypophysiotropic hormones occurred in arthritic rats. Melatonin treatment prevented the alteration in 24-hour rhythms of serum ACTH, prolactin and luteinizing hormone in rats injected with FCA. In addition, melatonin pretreatment prevented the alteration in the 24-hour variation in hypothalamic serotonin and dopamine turnover during the preclinical phase of Freund's adjuvant arthritis in rats. Some pinealectomy-induced immune changes in arthritic rats were also prevented by physiological concentrations of melatonin. Melatonin may play the role of an 'internal synchronizer' for the immune system. PMID- 14739558 TI - Translational activity of N-methyl-D-aspartate receptor subunit NR1 mRNA in PC12 cells. AB - PC12 cells contain NR1 mRNA but lack significant expression of NR1 protein suggesting translational or posttranslational regulation. Translational activity of NR1 mRNA in PC12 cells was examined by sucrose gradient fractionation and by heterologous luciferase NR1 gene expression studies. The cosedimentation and association of NR1 mRNA with large polyribosomes (polysomes) confirmed the translatability of NR1 message in PC12 cells. Possible initiation and/or elongation defects during the translation of NR1 mRNAs were investigated by cyclohexamide treatment. The marked decline in the number of ribosomes associated with NR1 mRNA after prolonged exposure to cyclohexamide suggested that initiation was limiting translation of NR1 mRNA in PC12 cells. Consequently, the effect of the 5' and 3' untranslated regions (UTRs) on translation was examined using fusion constructs consisting of the luciferase coding region fused to either or both the 5' UTR and 3' UTR of NR1. The transfection of PC12 cells with the luciferase NR1-UTR fusion constructs revealed that the 3' UTR of NR1 had a significant inhibitory effect on luciferase expression. In contrast, the 5' UTR of NR1 had no inhibitory effect on mRNA translation in PC12 cells. The results from this study indicate that the translation of NR1 mRNA in PC12 cells may be impeded at initiation and this inhibition may be regulated at least in part through the 3' UTR of NR1. PMID- 14739559 TI - Differential roles of spinal protein kinases C and a in development of primary heat and mechanical hypersensitivity induced by subcutaneous bee venom chemical injury in the rat. AB - It has been demonstrated that subcutaneous injection of bee venom (BV) can produce different types of pain and hypersensitivity including persistent spontaneous nociception (PSN), primary heat and mechanical hypersensitivity (hyperalgesia) and mirror-image heat (MIH) hypersensitivity in an individual animal, and the changes of spinal neurons are likely to be responsible for the production of these pain-related behaviors. In this study, we examined the roles of spinal protein kinase C (PKC) and protein kinase A (PKA) in the BV-induced different types of pain and hypersensitivity in conscious rats. We found that: (1). BV-induced primary heat hypersensitivity could be blocked by intrathecal pre or posttreatment with a PKC inhibitor, chelerythrine chloride (CH), while a PKA inhibitor, N-(2-[P-bromocinnamylamino]ethyl)-5-isoquinolinesulfonamide hydrochloride (H89), had no effect. (2). BV-induced primary mechanical hypersensitivity could be blocked by pre- or posttreatment with H89, whereas CH had no effect. (3). Both pre- and posttreatment with H89 produced suppressive effects on both induction and maintenance of the BV-induced PSN and MIH hypersensitivity. Based on the present findings, we proposed that spinal PKC might be activated during the central processes of primary heat hypersensitivity, while spinal PKA is likely to be involved in primary mechanical hypersensitivity induced by subcutaneous BV chemical injury. Taken together with our previous report however, spinal PKC and PKA are likely to be simultaneously involved in the central processes of both PSN and MIH hypersensitivity. PMID- 14739560 TI - Differential expression of alpha-amino-3-hydroxy-5-methyl-4-isoxazole-propionate glutamate receptors in the rat striatum during postnatal development. AB - Alpha-amino-3-hydroxy-5-methyl-4-isoxazole-propionate (AMPA)-type glutamate receptors (GluR1-4) are one of the most important ionotropic glutamate receptors in the striatum, a key region of the basal ganglia. The present study investigated the trend of developmental expression of AMPA receptor subunits in the striatum of rats in different developmental stages, i.e., at postnatal day 7 (PND7), PND21 and adult. Perfuse-fixed striatal sections were used. The expression of AMPA subunits was studied by immunofluorescence and reverse transcriptase-polymerase chain reaction (RT-PCR). RT-PCR revealed that the levels of expression of the GluR1 and GluR3 mRNAs were the same among the age groups. The level of GluR2 mRNA was highest in PND21 rats and lowest in adult. The highest level of GluR4 mRNA was detected in rats at PND7. Similar trends of GluR1, GluR2 and GluR2/3 immunoreactivity expression were observed using commercially available specific antibodies. In addition, a large proportion of parvalbumin-immunoreactive GABAergic interneurons in the striatum were found to display GluR1 immunoreactivity in PND21 and adult rats. In contrast, most of the choline acetyltransferase-immunoreactive cholinergic interneurons were found to display GluR2 immunoreactivity but less GluR1 and no GluR2/3 immunoreactivity in PND21 and adult rats. The present study suggests that there is a distinct pattern of expression of AMPA-type receptor mRNAs and proteins in the rat striatum at different stages of development. PMID- 14739561 TI - Lack of evidence of an interaction between leu-enkephalin and muscarinic-like responses in the frog semicircular canal. AB - Using multiunit recording of action potentials from the whole vestibular nerve, we studied whether opioid peptide leu-enkephalin (Enk) may modulate muscarinic like responses in semicircular organs of the frog. When acetylcholine (ACh) (0.1 1 mM) was applied with 1 nM Enk, the maximal frequency increase induced by ACh was reduced.However,the frequency decrease of ACh responses under Enk did not differ from the frequency decline in basal spike discharge induced by Enk alone. Administration of atropine (1 microM) left the response to Enk intact and blocked the excitatory effect of ACh. No modification of the ACh response under Enk was observed in the presence of the non-selective opioid receptor antagonist naloxone (10 microM). This study suggests that no interaction exists between the ACh mediated excitatory action on resting activity in the isolated semicircular canal preparation and the suppressive action of Enk. PMID- 14739562 TI - Salsolinol causing parkinsonism activates endoplasmic reticulum-stress signaling pathways in human dopaminergic SK-N-SH cells. AB - The endoplasmic reticulum (ER) is a small intracellular organelle to which one third of cellular proteins are translocated after translation and post translational modification, folding and the formation of a three- or four dimensional structure. ER also has a role in the transportation of proteins to other intracellular organelles, the cell surface or the outer space of the cell membrane. Thus, ER is an important intermediate which maintains intracellular homeostasis through complex control systems. Once these control systems are disrupted, serious disturbances occur. Many neurodegenerative diseases including Parkinson's disease involve aggregation and deposition of misfolded proteins such as alpha-synuclein. Endogenously occurring neurotoxins such as Salsolinol and 1 benzyl-1,2,3,4-tetrahydroisoquinoline (1BnTIQ) causing Parkinsonism may foster misfolded proteins and bring forth ER stress in dopaminergic neurons. In the present study we examined translational changes fostered by ER stress and mediated by the Parkinsonian endogenous neurotoxins, salsolinol and 1BnTIQ, in dopaminergic cell line. Treatment with salsolinol and 1BnTIQ induced several genes involved in ER stress and unfolded protein response (UPR), such as ER chaperones and GADD153 (CHOP). Immunoblotting confirmed phosphorylation of the key endoplasmic reticulum stress kinase PERK (PKR-like-ER kinase) and eIF2alpha and induction of their downstream targets such as Bip and GADD153. These findings suggest a widespread involvement of ER stress and unfolded protein response in the pathophysiology of Parkinson's disease. PMID- 14739563 TI - A systematic review of several potential non-genetic risk factors for multiple sclerosis. AB - We reviewed the literature published in the English language to determine the weight of evidence for several potential non-genetic risk factors for multiple sclerosis, including solar ultraviolet radiation (UVR), sex hormones and dietary fat/fatty acids. We ranked the plausibility of each factor and graded the methodological rigour of case-control and cohort studies to determine whether there was a sufficient number of high-quality studies to weigh the evidence. Based on our criteria, the plausibility for solar UVR and sex hormones is good and fair for dietary fat/fatty acids. However, the body of epidemiologic evidence is insufficient for these three sets of risk factors. We did not find a sufficient number of methodologically rigorous studies to weigh the evidence for any of the potential risk factors we examined. PMID- 14739564 TI - Comparison of environmental and genetic factors for Parkinson's disease between Chinese and Caucasians. AB - This review paper compares the differences in prevalence, and environmental and genetic risk factors for Parkinson's disease between Chinese and Caucasian subjects. Comparison of age-specific prevalence between Chinese people and Caucasians suggests that the prevalence is lower in the Chinese (at least in the past), although the prevalence rate in China appears to be rising. Distinctions in environmental risk factors and genetic factors are discussed. The difference in prevalence may be due to distinctions in environmental and genetic risk factors as well as the complex interaction between these environmental and genetic factors, although discrepancies in methodology for prevalence surveys can also be an explanation. PMID- 14739565 TI - Association between symptoms reported in a population questionnaire and future ischemic stroke: the ARIC study. AB - The association between computer diagnosis derived from a symptom questionnaire and onset of first hospitalized ischemic stroke during follow-up for up to 11 years has been assessed for 11,804 participants in the Atherosclerosis Risk in Communities Study who had no baseline history of stroke. Of these participants, 578 reported prior positive transient ischemic attack/stroke symptoms, and 265 strokes occurred during the years 1987-1998. Adjusted for age, locale, sex, and race, persons with self-reported baseline symptoms had 2.8 times the hazard rate for incident ischemic stroke of those without symptoms, with 95% confidence interval 1.9-4.1. Greater relative risk was found among younger individuals, women, African-Americans, persons not current smokers, and those with lower white blood cell count. PMID- 14739566 TI - Cohort study of exposure to environmental tobacco smoke and risk of first ischemic stroke and transient ischemic attack. AB - The independent effect of exposure to environmental tobacco smoke (ETS; passive smoking) on the risk of stroke is not well established. We performed a cohort study among 27,698 lifelong nonsmokers with no prior history of stroke, 62% women, aged 30-85 years at enrollment (1979-1985). Self-reported ETS exposure at home and outside home (in hours/week) and stroke risk factors were collected at a health plan in San Francisco and Oakland. Follow-up for hospitalization and death was available through the end of 2000 (median = 16 years). In multivariate analysis adjusting for age, race/ethnicity, educational attainment, marital status, hypertension, diabetes and serum total cholesterol, ETS exposure at home of 20 h or more/week (in relation to <1 h/week) was associated with a 1.29-fold (95% CI 0.75-2.20) and a 1.50-fold (95% CI 1.07-2.09) increased risk of first ischemic stroke among men and women, respectively. No significant associations were found between ETS exposure outside home and ischemic stroke or between exposure to ETS at home or out of home and the risk of transient ischemic attack. Although potentially important confounders (such as dietary habits) were not included in the analysis, high-level ETS exposure at home was independently associated with increased risk of first ischemic stroke among never-smoking women. PMID- 14739567 TI - Cognitive impairment and dementia 20 months after stroke. AB - BACKGROUND AND PURPOSE: Dementia is common after stroke, but the dementia syndrome does not cover the whole spectrum of cognitive impairment. Our aim was to quantify and compare dementia and cognitive impairments in elderly patients 1.5 years after stroke and a matched normal population. SUBJECTS AND METHODS: We examined dementia and cognitive impairments in 149 out of an initial total of 243 acute stroke patients after a mean 20 months. Inclusion criteria were age > or =70 years, not living in an institution and no previous cerebral lesion. The patients' mean age was 81 years. Five controls matched by age and gender and fulfilling the same exclusion criteria were selected for each patient (n = 745) from a population-based survey in the same area. Dementia was diagnosed according to the DSM-III-R criteria, and impairments in different dimensions of cognitive function were assessed. RESULTS: The prevalence of dementia was 28% in the stroke patients and 7.4% in the controls (OR 4.7; 95% CI 3.0-7.5). Seventy-two percent of the patients had cognitive impairments compared to 36% in the controls. Cognitive impairments were more common in nondemented stroke patients than in nondemented controls: 61 vs. 31% (OR 3.5; 95% CI 2.3-5.3). The risk increase attributable to stroke was highest for patients below 80 years of age. CONCLUSIONS: Stroke confers an increased risk of dementia and cognitive impairments in the elderly, especially in the younger ones. Apraxia is the most frequent neuropsychiatric impairment after stroke. The concept of dementia does not describe cognitive impairments well, since it underestimates their extent not only after stroke but also in normal ageing. PMID- 14739568 TI - A follow-up analysis of transient ischemic attack patients suggests unsatisfactory disease management and possible underutilization of carotid endarterectomy in Lazio, Italy. AB - We evaluated the disease management of transient ischemic attack in patients admitted to Lazio hospitals from July 1997 to June 1998. We assessed the effects of patient characteristics including chronic comorbidities on the use of diagnostic procedures, endarterectomy, and on the risk of adverse cerebrovascular outcome or death. There were 2,608 patients in the study who were followed up over a 18- to 30-month period. Carotid surgery was performed on 1.15% of the subjects, total mortality was 34.7 per 1,000 person-years and adverse cerebrovascular outcome was observed in 38.1 per 1,000 person-years. Chronic comorbidities did affect the mortality rate and the rate of adverse outcome, but not the rate of endarterectomies. Carotid surgery was infrequently performed in study subjects. It seems that this potentially stroke-preventive treatment was not offered to suitable candidates in many instances. PMID- 14739569 TI - Attrition and non-compliance in secondary stroke prevention trials. AB - BACKGROUND: Attrition and non-compliance of subjects in secondary stroke prevention trials due to study drug-induced adverse events and loss to follow-up could lead to bias and loss of information, thus affecting the analysis of study results. METHODS: We reviewed results from ten antiplatelet stroke prevention clinical trials: CAN TIA, DUTCH TIA, SWED ASA, SALT, UK TIA, CATS, TASS, ESPS, ESPS-2, and CAPRIE to tabulate the frequencies for total subject discontinuation, voluntary withdrawal, and loss to follow-up. RESULTS: Forty thousand seven hundred and thirty (40,730) subjects participated in the aforementioned secondary stroke prevention trials. The range of outcomes was 11.8-52.0% for subjects discontinued for any reason (n = 9 trials); 3.0-20.9% for study drug-induced adverse events (n = 9 trials), and 4.2-7.8% for voluntary withdrawal (n = 10 trials). CONCLUSION: There is a substantial discrepancy (up to 20%) between the frequencies of total subject discontinuation for any reason and the sum of study drug-induced adverse events, voluntary withdrawal and loss to follow-up. Underestimation of these important outcomes may limit the ability of clinicians to translate results from clinical trials into medical practice. PMID- 14739570 TI - Validity and reliability of the 'Ten Questions' questionnaire for detecting moderate to severe neurological impairment in children aged 6-9 years in rural Kenya. AB - BACKGROUND: The 'Ten Questions' Questionnaire (TQQ) is used to detect severe neurological impairment in children living in resource-poor countries. Its usefulness has been established in Asia and the Caribbean, but there are a few published studies from Africa. We evaluated the TQQ as part of a larger study of neurological impairment in a rural community, on the coast of Kenya. METHODS: The study was conducted in two phases from June 2001 to May 2002; in phase one, a community household screening of 10,218 children aged 6-9 years using the TQQ was performed. Phase two involved a comprehensive clinical and psychological assessment of all children testing positive on the TQQ (n = 810) and an equivalent number of those testing negative (n = 766). Data were interpreted using the impairment-specific approach. RESULTS: Overall, the sensitivity rates for screening the different impairments were: cognitive (70.0%), motor (71.4%), epilepsy (100%), hearing (87.4%) and visual (77.8%). All the specificity rates were greater than 96%. However, the positive predictive values were low, and ranged from 11 to 33%. CONCLUSIONS: These results are similar to those from other continents and provide evidence that the TQQ can be used to compare the epidemiology of moderate/severe impairment in different parts of the world. Furthermore, the TQQ can be used to screen for moderately/severely impaired children in resource-poor countries; however, the low positive predictive values mean that other assessments are required for confirmation. PMID- 14739571 TI - Accuracy of death certificates for amyotrophic lateral sclerosis varies significantly from north to south of Italy: implications for mortality studies. AB - OBJECTIVE: To evaluate the accuracy of death certificates (DCs) for amyotrophic lateral sclerosis (ALS) in different parts of Italy. Studies based on DC diagnosis for ALS have shown a reduced mortality comparing northern with southern Italy. These data are in contrast with results from other surveys on the incidence of ALS performed in Italy and other countries. METHODS: Archives of neurological clinics from northern (Milano, Monza, Pavia, and Bologna) and southern Italy including islands (Napoli, Sassari, Palermo, and Messina) were searched for patients discharged with a diagnosis of ALS in the period 1970-1995. Subjects affected by definite/probable ALS according to the Scottish Motor Neuron Disease Research Group diagnostic criteria were included. DCs were obtained from the vital statistic bureau. True positive rates (TPRs) and 95% confidence intervals (CIs) for proportions were calculated for northern and southern Italy separately. Multiple logistic regression analysis was performed according to gender, age at onset, age and year of death, and interval between onset and death. RESULTS: We found 651 patients affected by definite/probable ALS; 573 of them had died by December 31, 1996. DCs were available for 566 subjects (411 from northern Italy and 155 from southern Italy). TPR was 66.7% (95% CI 61.9-71.2) for northern Italy and 51.6% (95% CI 43.5-59.7) for southern Italy (chi(2) = 10.9, p = 0.001). Logistic regression analysis showed an association between a lower accuracy of DCs and the interval between onset of symptoms and death. TPR calculations considering different death periods (1970-1982 and 1983-1996) showed comparable rates of accuracy over time. CONCLUSIONS: Mortality statistics based on official death records do not accurately reflect interregional mortality for ALS in Italy. PMID- 14739572 TI - A meta-analysis of maternal cured meat consumption during pregnancy and the risk of childhood brain tumors. AB - OBJECTIVE: N-Nitroso compounds (NOCs) are recognized neural carcinogens in animal models and are suspected human carcinogens. A meta-analysis was performed examining the possible association of maternal intake of cured meat (an important source of dietary NOCs) during pregnancy and the risk of pediatric brain tumors. METHODS: Data from epidemiological studies were pooled using a general variance based meta-analytic method employing confidence intervals described by Greenland in 1986. The outcome of interest was a summary relative risk (RR) reflecting the risk of childhood brain tumor (CBT) development associated with maternal intake of cured meats during pregnancy. Sensitivity analyses were performed when necessary to explain any observed statistical heterogeneity. RESULTS: Seven observational studies were found that met the protocol-specified inclusion criteria. Analysis for heterogeneity demonstrated a lack of statistical heterogeneity (p = 0.59), indicating that the data could be statistically combined. Pooling data from the 6 reports containing data on maternal cured meat intake of all types yielded an RR of 1.68 (1.30- 2.17), being a statistically significant result. Analyzing CBT risk by type of cured meat ingested showed that hot dog consumption increased CBT risk by 33% (1.08-1.66), with a similar increase shown by frequent ingestion of sausage, i.e. 44%. CONCLUSION: The data provide support for the suspected causal association between ingestion of NOCs from cured meats during pregnancy and subsequent CBT in offspring. Limitations in study design preclude definitive conclusions, but the relationship warrants exploration via additional observational and laboratory-based studies. PMID- 14739573 TI - Consistency of primary brain tumor diagnoses and codes in cancer surveillance systems. AB - High-quality cancer registry data are essential for assessing trends in incidence rates. This study evaluated the consistency of brain tumor surveillance data using a random sample of cases from the Connecticut Tumor Registry. Three neuropathologists independently and blindly reviewed tumor slides from 204 cases and a nosologist blindly reviewed and assigned International Classification of Diseases for Oncology (ICD-O) codes to 326 cases. For the pathology review, absolute concordance was as high as 81% for all primary brain tumors. Absolute concordance rates were high for nerve sheath (89%), meningioma (95%), and pituitary (95%) tumors. Rates were much lower for malignant tumors. ICD-O coding of malignant brain tumors is of relatively high quality with the exception of mixed gliomas and unspecified tumors. A high level of consistency for nonmalignant brain tumor diagnoses suggests that rates for these tumors, when actively reported to a surveillance system, can be of high quality. PMID- 14739574 TI - Association between statin use and Alzheimer's disease. AB - CONTEXT: 3-Hydroxy-3-methylglutaryl coenzyme A reductase inhibitors (statins) have been associated with a decreased risk for Alzheimer's disease (AD). OBJECTIVE: To evaluate the association between statin use and AD adjusted for comorbid medical conditions. DESIGN: A nested case-control study. PATIENTS: Patients at the Veterans Affairs Medical Center in Birmingham, Ala., USA with a new diagnosis of AD (cases) between 1997 and 2001 (n = 309) and age-matched non AD controls (n = 3,088). MAIN OUTCOME MEASURE: Odds ratio for association between AD and statin use. RESULTS: Statin users had a 39% lower risk of AD relative to nonstatin users (odds ratio 0.61, 95% confidence interval 0.42-0.87). This association appeared to be modified by the presence of certain chronic medical conditions (i.e., hypertension, ischemic heart disease and cerebrovascular disease) in that the reduced risk was observed among those with these diseases, whereas no association was observed among those without any of these conditions. CONCLUSIONS: In this study, following adjustment for confounding factors, a statistically significant inverse association between statin use and AD was observed. The results lend support to looking at AD outcomes in trials of statins to further evaluate their possible beneficial effects. PMID- 14739575 TI - Use of the capture-recapture method for determining the prevalence of neurological parasitic diseases. PMID- 14739577 TI - Relationship of pregnancy-associated plasma protein-a to renal function and dialysis modalities. AB - BACKGROUND: The aim of the study was to determine pregnancy-associated plasma protein-A (PAPP-A), which was recently described as a new marker of cardiovascular events, in patients with chronic renal insufficiency/failure and to find out its relationship to renal function and to prominent markers of oxidative stress (advanced oxidation protein products--AOPP) and inflammation (C reactive protein--CRP). METHODS: The studied group consisted of 36 chronic hemodialysis patients (HD), 10 patients treated with continuous ambulatory peritoneal dialysis (CAPD) and 38 patients with chronic renal insufficiency (CHRI) not yet dialyzed. PAPP-A was measured by Time Resolved Amplified Cryptate Emission technology. Determination of AOPP is based on a spectrophotometric method. RESULTS: PAPP-A levels are statistically significantly elevated in the both groups of dialyzed patients in comparison with healthy subjects (27.0 +/- 16.5 mIU/l in HD and 14.07 +/- 6.73 mIU/l in CAPD vs. 8.22 +/- 2.7 mIU/l in the control group, p < 0.0001 and p < 0.001, respectively, p < 0.05 HD vs. CAPD). The mean serum PAPP-A levels in the CHRI patients not yet dialyzed were not significantly higher in comparison with the control group (9.72 +/-4.44 vs. 8.22 +/- 2.7 mIU/l, n.s.). In the CHRI not dialyzed patients, we found a significant positive correlation between serum creatinine and PAPP-A levels (r = 0.68, p < 0.05). In comparison with controls, AOPP and CRP levels were significantly higher in HD patients [AOPP 155.0 +/- 37.9 micromol/l, p < 0.0001 vs. controls, CRP 10.0 (4.6- 26.9) mg/l (median, interquartile range), p < 0.0001 vs. controls], CAPD patients [AOPP 118.5 +/- 25.8 micromol/l, p < 0.0001 vs. controls, CRP 7.7 (2.0 18.8) mg/l, p < 0.01 vs. controls] and AOPP levels in chronic renal failure patients not yet dialyzed (98.5 +/- 43.24 micromol/l, p < 0.01 vs. controls). The correlations between PAPP-A and AOPP (r = 0.49, p < 0.05) and PAPP-A and CRP (r = 0.48, p < 0.05) serum concentration were statistically significant in HD patients. In CAPD patients, neither a correlation between PAPP-A and AOPP nor a correlation between PAPP-A and CRP were found. CONCLUSION: We can conclude that serum PAPP-A levels sensitively reflect the changes in renal function, depend on dialysis modality, and may represent a novel marker associated with inflammation and oxidative stress in chronic renal failure patients. PMID- 14739578 TI - Statistical evaluation of local alignment features predicting allergenicity using supervised classification algorithms. AB - BACKGROUND: Recently, two promising alignment-based features predicting food allergenicity using the k nearest neighbor (kNN) classifier were reported. These features are the alignment score and alignment length of the best local alignment obtained in a database of known allergen sequences. METHODS: In the work reported here a much more comprehensive statistical evaluation of the potential of these features was performed, this time for the prediction of allergenicity in general. The evaluation consisted of the following four key components. (1) A new high quality database consisting of 318 carefully selected, non-redundant allergens and 1,007 sequences carefully selected to be non-allergens. (2) Three different supervised algorithms: the kNN classifier, the Bayesian linear Gaussian classifier, and the Bayesian quadratic Gaussian classifier. (3) A large set of local alignment procedures defined using the FASTA3 alignment program by means of a wide range of different parameter settings. (4) Novel performance curves, alternative to conventional receiver-operating characteristic curves, to display not only average behaviors but also statistical variations due to small data sets. RESULTS: The linear Gaussian classifier proved most useful among the tested supervised machine learning algorithms, closely followed by the quadratic Gaussian equivalent and kNN. The overall best classification results were obtained with a novel feature vector consisting of the combined alignment scores derived from local alignment procedures using different substitution matrices. CONCLUSIONS: The models reported here should be useful as a part of an integrated assessment scheme for potential protein allergenicity and for future comparisons with alternative bioinformatic approaches. PMID- 14739579 TI - Ginsenoside Rh1 possesses antiallergic and anti-inflammatory activities. AB - BACKGROUND: Ginseng (the root of Panax ginseng C.A. Meyer, Araliaceae) has been reported to possess various biological activities, including anti-inflammatory and antitumor actions. In this study, we investigated the antiallergic activity of ginsenosides isolated from ginseng. METHOD: We isolated ginsenosides by silica gel column chromatography and examined their in vitro and in vivo antiallergic effect on rat peritoneal mast cells and on IgE-induced passive cutaneous anaphylaxis (PCA) in mice. The in vitro anti-inflammatory activity of ginsenoside Rh1 (Rh1) in RAW264.7 cells was investigated. RESULTS: Rh1 potently inhibited histamine release from rat peritoneal mast cells and the IgE-mediated PCA reaction in mice. The inhibitory activity of Rh1 (87% inhibition at 25 mg/kg) on the PCA reaction was found to be more potent than that of disodium cromoglycate (31% inhibition at 25 mg/kg); Rh1 was also found to have a membrane-stabilizing action as revealed by differential scanning calorimetry. It also inhibited inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) protein expression in RAW 264.7 cells, and the activation of the transcription factor, NF kappaB, in nuclear fractions. CONCLUSION: The antiallergic action of Rh1 may originate from its cell membrane-stabilizing and anti-inflammatory activities, and can improve the inflammation caused by allergies. PMID- 14739580 TI - Parietaria profilin shows only limited cross-reactivity with birch and grass profilins. AB - BACKGROUND: In patients showing multiple reactivity to seasonal airborne allergens, a skin prick test (SPT) with Parietaria pollen scores frequently negative, suggesting that profilin from Parietaria might not share IgE-binding epitopes with profilin from botanically unrelated airborne allergens. This study investigated the immunologic cross-reactivity between profilins from Parietaria, grass and birch pollen. METHODS: 36 patients hypersensitive to birch profilin, Bet v 2, underwent SPT with Parietaria, and IgE to both whole Parietaria pollen and Phleum profilin were sought in their sera by ELISA. In ELISA inhibition studies, IgE reactivity to Phleum profilin was measured before and after absorption of sera from Parietaria reactors with both whole Parietaria pollen extract, and Par j 1/Par j 2 fraction. Further, their IgE reactivity to whole Parietaria pollen was measured before and after absorption with the Par j 1/Par j 2 fraction. RESULTS: All sera showed IgE reactivity to Phleum profilin. Only 14/36 (39%) patients had a positive SPT to Parietaria and 17/36 (47%) showed IgE to Parietaria. Absorption of sera from Parietaria reactors with whole Parietaria extract caused a marked reduction in IgE reactivity to grass profilin; in contrast, absorption of sera with the Par j 1/Par j 2 fraction did not exert any inhibitory effect on IgE reactivity to grass profilin. Absorption of sera with the Par j 1/Par j 2 fraction markedly reduced IgE reactivity to whole Parietaria extract in 8/9 cases. CONCLUSION: Less than 50% of patients sensitized to birch and grass profilin cross-react to Parietaria profilin. In most cases, cross reactors are hypersensitive to major, specific Parietaria allergens as well. This findings may be of practical relevance when the prescription of specific immunotherapy is considered. PMID- 14739581 TI - Zebrafish as an experimental model: strategies for developmental and molecular neurobiology studies. AB - Zebrafish provide a rapid and effective means for assessing gene function in the vertebrate nervous system. By employing gain- and loss-of-function techniques it is possible to obtain insights into the roles of both wild-type and heterologously expressed genes. Such approaches enable rapid progression from gene discovery to gene expression and finally to gene function even when examining development of a tissue as complex as the nervous system. Exploiting the full potential of zebrafish as a bioassay for the nervous system will require, not only an understanding of the molecular and cellular basis of normal zebrafish development, but also an appreciation of comparative processes in other species. When applied to mutant animals, classic morphological approaches and contemporary molecular genetic techniques are providing a wealth of information on the development of the nervous system at the molecular, cell, system and behavioural levels. Zebrafish are now emerging as an important tool, supporting mouse genetical approaches for understanding neural function in vertebrates. PMID- 14739582 TI - Neural cell fate analysis in zebrafish using olig2 BAC transgenics. AB - We describe here simple methods for producing transgenic zebrafish reporter lines using BAC clones. The use of BAC clones facilitates creation of useful transgenics as the large amounts of genomic DNA they contain increase the likelihood that reporter gene expression will be properly regulated. Combined with recent advances in live embryo image analysis, this strategy has the potential to greatly advance the investigation of neural cell behavior during development. PMID- 14739583 TI - Neuron-specific gene manipulations to transparent zebrafish embryos. AB - To investigate the molecular basis of neural network formation, we introduced a novel double-cassette vector approach for visualizing and manipulating neuronal development in living zebrafish embryos. Two genes are physically linked in the double-cassette vector system, which ensures co-expression of an effector-protein and an EGFP-reporter in the same neuron. By generating transgenic enhanced green fluorescent protein (EGFP) expressing zebrafish lines, we first established that EGFP under control of either the olfactory marker protein (OMP) gene promoter or the nicotinic acetylcholine receptor beta3 (nAChRbeta3) gene promoter, directed strong EGFP expression to the olfactory sensory neurons and the retinal ganglion cells (RGCs), respectively. These transgenic lines allowed the visualization of the development of the entire olfactory sensory neurons and RGCs in vivo. By injection of vectors with EGFP under control of either the OMP or the nAChRbeta3 gene promoter, we followed the development of individual olfactory sensory neurons and RGCs. The double-cassette expression vector strategy enabled us to clarify the roles of protein kinase A (PKA) and glycogen synthase kinase-3beta (GSK-3beta) in the development of olfactory sensory neurons and RGCs. The combination of visualization and neuron-specific gene manipulation provides a powerful reverse genetic in vivo approach for the study of genes of interest in neural differentiation, axonal pathfinding, and synaptogenesis. PMID- 14739584 TI - Practical procedures for ectopic induction of gene expression in zebrafish embryos using Bhc-diazo-caged mRNA. AB - We previously reported mRNA caging technology as a novel and simple technique for photo-mediated temporal and spatial control of gene activation in zebrafish embryos and as an alternative to the 'gene knockdown' approach using antisense morpholino oligonucleotides. The caging reagent used is 6-bromo-4-diazomethyl-7 hydroxycoumarin (Bhc-diazo), which forms a covalent bond with the phosphate moiety of the sugar-phosphate backbone of RNA. Mainly because of the reduced solubility of caged mRNA in aqueous solutions, special care in handling is needed. The Bhc-diazo group binds to the phosphate moieties of RNA and abolishes the translational activity of the latter. The translational activity of Bhc-caged mRNA is restored by photolysis/uncaging when exposed to long-wave UV light (350 approximately 365 nm). In this paper we describe the technique and detailed procedures for spatially and temporally controlled induction of gene expression in zebrafish embryos. PMID- 14739585 TI - Identifying axon guidance defects in the embryonic zebrafish brain. AB - The method described here outlines a simple protocol to investigate the in vivo function of axon guidance molecules during the development of the embryonic zebrafish brain. By 24 hours postfertilization, a simple scaffold of axon tracts and commissures can be visualised in the brain using acetylated alpha-tubulin, a panaxonal marker that stains all axons. The highly stereotypical trajectory of axons in the embryonic zebrafish brain provides an ideal system in which to study the molecular mechanisms of axon guidance, as defects in the axon scaffold can be clearly visualised. We describe here our approach to identify defects in the trajectory of axons that establish the initial template of tracts in the embryonic fore- and mid-brain. By combining immunohistochemical techniques and confocal microscopy on dissected wholemounts of embryonic brains we are able to observe at high resolution the complete scaffold of axon tracts. This approach provides a rapid and simple means of assessing axon guidance defects in the developing brain. Given the advantages of the zebrafish as a model system, and the range of molecular perturbation methods now available, this technique provides a valuable tool for assessing the phenotypic effects of gene perturbations in a biologically relevant context. PMID- 14739587 TI - Zebrafish retinal slice preparation. AB - This paper describes the protocol for generating thin (approximately 100 microm) slices of the zebrafish retina. Retinal slices retain the cytoarchitecture and synaptic contacts found in vivo, allowing neurons to be identified prior to physiological recordings. These characteristics distinguish retinal slices from both isolated cell and eyecup preparations. Studies using the zebrafish retinal slice have classified different retinal cell types, documented voltage- and ligand-gated current responses in distal bipolar neurons, and correlated physiological responses with neuronal morphology. Data collected using this protocol have provided baseline information about retinal circuitry that can be directly applied to behavioral studies examining visual function and/or mutants with visual system defects. PMID- 14739586 TI - Assessment of neuronal maturation and acquisition of functional competence in the developing zebrafish olfactory system. AB - Olfactory coding at the level of the olfactory bulb is thought to depend upon an ensemble response of mitral cells receiving input from chemotopically-organized projections of olfactory sensory neurons and regulated by lateral inhibitory circuits. Immunocytochemical methods are described to metabolically classify neurons in the developing zebrafish olfactory system based on the relative concentrations of taurine, glutamate, GABA (and potentially other small biogenic amines) and a small guanidium-based cation, agmatine, which labels NMDA-sensitive cells by permeating through active ionotropic glutamate receptor channels. Using metabolic profiling in conjunction with activity dependent labeling we demonstrate that neuronal differentiation in the developing olfactory bulb, as assessed by acquisition of a mature neurochemical profile, and sensitivity to an ionotropic glutamate receptor agonist, NMDA, occurs during the second day of development. This experimental approach is likely to be useful in studies concerned with the development of glutamatergic signaling pathways. PMID- 14739588 TI - Whole-cell patch-clamp recordings from identified spinal neurons in the zebrafish embryo. AB - We describe a preparation for obtaining patch-clamp recordings from identified embryonic spinal cord interneurons, motoneurons and sensory neurons in an in vivo zebrafish preparation. This preparation is used to study the spatial and temporal patterns of spontaneous and touch-evoked electrical activity during the initial development of circuitry in the spinal cord. The combination of these physiological techniques with the powerful genetic and molecular tools available in the zebrafish has the potential to increase our understanding of the complex interactions between genes and electrical activity during the development of the vertebrate nervous system. PMID- 14739589 TI - Double labeling of neurons by retrograde axonal tracing and non-radioactive in situ hybridization in the CNS of adult zebrafish. AB - A number of genes affecting axonal projections are currently being identified in zebrafish mutant screens. Analyzing the expression of these genes in the adult brain in relation to specific neuronal populations could yield insights into new functional contexts, such as the successful axonal regeneration in adult zebrafish. Here, we provide a relatively simple procedure for non-radioactive in situ hybridization in sections of adult zebrafish brains in combination with retrograde axonal tracing using the fluorescent neuronal tracer rhodamine dextran amine (RDA). A lesion is inflicted on the spinal cord of adult zebrafish and a crystal of RDA is then applied to the lesion site resulting in retrograde labeling of neurons in the brain through their spinal axons. Six to eighteen days later fish are perfusion-fixed, and in situ hybridization is carried out on vibratome-cut floating sections using a protocol simplified from that used for whole-mounted zebrafish embryos. This procedure leads to robust double labeling of axotomized neurons with RDA and an in situ hybridization signal for the growth associated protein 43 (GAP-43). This method can be used to identify gene expression in specific populations of projection neurons and to detect changes in gene expression in axotomized neurons in the CNS of adult zebrafish. PMID- 14739590 TI - Using the adult zebrafish visual system to study cadherin-2 expression during central nervous system regeneration. AB - Zebrafish is a good model organism for the study of vertebrate development due to numerous experimental advantages. Here we show that zebrafish are also suitable for the study of cellular and molecular mechanisms underlying tissue regeneration in the vertebrate central nervous system. In this report we describe simple protocols to introduce injuries to the adult zebrafish visual structures and analyze cadherin-2 expression in the regenerating visual system using immunocytochemistry and immunoblotting. PMID- 14739591 TI - Immunocytochemistry as a tool for zebrafish developmental neurobiology. AB - Two methods are presented here that allow clear visualization of antibody localization in zebrafish whole mount preparations, both for immunocytochemistry (ICC) alone and in combination with in situ hybridization (ISH). The first protocol describes ICC performed using a modified permeabilization technique and the chromogen AEC (3-Amino-9-ethylcarbazole). The second protocol describes the co-localization of transcriptional and translational products using a combined ISH/ICC protocol. A fluorescing chromogen (Fast Red, FR) is used to detect mRNA transcripts by ISH, and is combined with ICC that uses a secondary antibody conjugated to a different fluorescent molecule (Alexa 488). These procedures allow the identification of gene expression patterns in cell types identifiable with known antibodies. PMID- 14739592 TI - Comparing protein stabilities during zebrafish embryogenesis. AB - The stabilities of many key proteins are regulated, e.g. via ubiquitination and proteasomal degradation, with important biological consequences. We present a convenient method that allows the analysis and comparison of protein stabilities during embryogenesis using early zebrafish development as a model system. Basically, this method involves ectopic overexpression of epitope-tagged proteins via mRNA injections in one-to-four-cell stage embryos and subsequent protein detection after various time points. Indeed, the protein stability of the ubiquitin ligase RLIM, which is able to autoubiquitinate and target itself for proteasomal degradation, was much shorter when compared to a protein consisting of a Myc epitope-tag and a nuclear localization domain. Thus, this method may be used more widely for the study of developmental protein stability. PMID- 14739594 TI - TMS1/ASC: the cancer connection. AB - TMS1/ASC is a bipartite protein comprising two protein-protein interaction domains, a pyrin domain (PYD) and a caspase recruitment domain (CARD). Proteins containing these domains play pivotal roles in regulating apoptosis and immune response pathways, and mutations in a number of PYD- and CARD-containing proteins have been linked to autoinflammatory diseases and cancer. Indeed, one of the ways in which TMS1/ASC was identified was as a target of methylation-mediated silencing in breast cancer cells. This review discusses the mounting evidence supporting a correlation between the silencing of TMS1/ASC expression and cancer. In addition, it addresses the reported functions of TMS1/ASC that include apoptosis, activation of inflammatory caspases and regulation of NF-kappa B, and discusses the potential ways in which loss of TMS1/ASC contributes to carcinogenesis. PMID- 14739595 TI - Induction of apoptosis by tumor cell-targeted toxins. AB - Targeted toxins are fusion proteins that combine a targeting molecule that selectively binds to and enters tumor cells with a protein toxin that kills the target cells. These molecules represent an exciting approach to develop effective cancer-specific therapeutics that have few side effects on normal tissues and numerous such toxins are in various stages of pre-clinical and clinical development to treat a wide variety of tumors. In this review, we discuss this strategy, describe ways that the toxins activate the apoptosis machinery and discuss future developments in this field. PMID- 14739596 TI - Antiapoptotic role of heme oxygenase (HO) and the potential of HO as a target in anticancer treatment. AB - Heme oxygenase-1 (HO-1) is an inducible enzyme that catalyzes oxidative degradation of heme to form biliverdin, carbon monoxide (CO), and free iron. Biliverdin is subsequently reduced to bilirubin by the enzyme biliverdin reductase. Increasing evidence has indicated the critical role of HO-1 in cytoprotection and more diverse biological functions. Induction of HO-1 by various chemical inducers that are primarily cell stress inducers or by HO-1 gene transfection confers a protective capacity to cultured cells as well as to cells in several in vivo animal models. In addition, HO-1-deficient mice exhibit a significant increase in susceptibility to tissue injury. The cytoprotective action of HO-1 seems to be mainly a function of the antiapoptotic effects of the enzyme. HO-1 is believed to exert this antiapoptotic action by multiple mechanisms: (a) decreased intracellular pro-oxidant levels, (b) increased bilirubin levels, and (c) elevated CO production. CO may produce an antiapoptotic effect by inhibiting both expression of p53 and release of mitochondrial cytochrome c. HO-1 may also be a target in antitumor therapy because the growth of most tumors depends on HO-1. Our preliminary studies with an HO inhibitor showed a promising antitumor effect. This preliminary work warrants continued investigation for possible novel anticancer chemotherapy. PMID- 14739597 TI - The dependence receptor hypothesis. AB - A new family of functionally-related receptors has recently been proposed, dubbed dependence receptors. These proteins, only some of which share sequence similarities, display the common property that they transduce two different intracellular signals: in the presence of ligand, these receptors transduce a positive signal leading to survival, differentiation or migration; conversely, in the absence of ligand, the receptors initiate or amplify a signal for programmed cell death. Thus cells that express these proteins at sufficient concentrations manifest a state of dependence on their respective ligands. The signaling that mediates cell death induction upon ligand withdrawal is in large part uncharacterized, but typically includes a required interaction with, and cleavage by, specific caspases. Here, we review the current knowledge concerning dependence receptors, including the shared mechanisms for cell death induction and their potential relevance in nervous system development and regulation of tumorigenesis. PMID- 14739598 TI - SiRNA based strategies for inhibition of apoptotic pathways in vivo--analytical and therapeutic implications. AB - Apoptosis is a genetically controlled mechanism of cell death which is important for embryogenesis, metamorphosis, tissue homeostasis and tumor regression of multicellular organisms. In normal cells as well as in transformed cells signals released from the cytoplasm and/or the cell membrane can trigger the activation of caspases which in turn cleave many cellular substrates, leading to the characteristic morphology of apoptosis. Systematic analysis and dissection of apoptotic pathways was obtained by the use of knock out or transgenic organisms, expressing dominant active or negative proteins. The use of antisense oligonucleotides (ASO) for analysis of apoptotic pathways was commonly restricted to cultured cells, because of the low efficacy of ASO in in vivo experiments. In order to investigate the contribution of specific apoptotic pathways in the onset and maintenance of disease in vivo experiments are needed. This approach allows the analysis of apoptotic pathways within their physiological/pathophysiological environment. The combination of recent advances in in vivo gene delivery with siRNA technique for efficient gene silencing provides new, unique possibilities to study apoptotic pathways thereby evaluating new molecular therapeutic strategies in vivo. In this minireview we will focus on the use of RNA interference for analytical and therapeutical suppression of apoptotic pathways in vivo with special consideration of the liver. PMID- 14739599 TI - Apoptosis in serum-deprived vascular smooth muscle cells: evidence for cell volume-independent mechanism. AB - Shrinkage is the earliest hallmark of cells undergoing apoptosis. This study examines the role of this phenomenon in the onset of vascular smooth muscle cell (VSMC) apoptosis triggered by growth factor withdrawal. In hyperosmotic media, VSMC showed the same amplitude of shrinkage but were more resistant to apoptosis than endothelial, epithelial and immune system cells. As with growth factor withdrawal, apoptosis in hyperosmotically-shrunken VSMC was sharply potentiated by transfection with E1A-adenoviral protein and was suppressed by activation of cAMP signaling as well as by the pan-caspase inhibitor z-VAD.fmk. Both cell shrinkage and apoptosis in VSMC-E1A treated with hyperosmotic medium were potentiated under sustained Na+, K+ pump inhibition with ouabain that was in contrast to inhibition of apoptosis documented in ouabain-treated, serum-deprived cells. After 1-hr incubation in serum-deprived medium, VSMC-E1A volume declined by approximately 15%. Transfer from hypotonic to control medium decreased VSMC E1A volume by approximately 25% without any induction of apoptosis. Neither swelling in hyposmotic medium nor dissipation of the transmembrane gradient of K+ and major organic osmolytes protected serum-deprived VSMC-E1A from apoptosis. Thus, our results show that similarly to immune system, endothelial and epithelial cells, extensive VSMC shrinkage in hyperosmotic medium leads to the development of apoptosis. In contrast to hyperosmotic medium, the modest cell volume decrease occurring in serum-deprived VSMC does not contribute to triggering of the apoptotic machinery. PMID- 14739601 TI - Stress-induced apoptosis: toward a symmetry with receptor-mediated cell death. AB - Apoptosis is a form of programmed cell death executed by caspases activated along signalling pathways initiated by ligation of cell-surface death receptors ( extrinsic pathway ) or by perturbation of the mithocondrial membrane promoted by physical or chemical stress agents ( intrinsic pathway ). In metazoans, this evolutionary conserved, genetically controlled process has a role in a variety of physiological settings, as development, homeostasis of tissues and maintenance of the organism integrity. When deranged by impaired regulation or inappropriate activation apoptosis contributes to the pathogenesis of diseases as autoimmunity, cancer, restenosis, ischaemia, heart failure and neurodegenerative disorders. In this review we will present a survey of the stress-induced intrinsic, mithochondrial, pathway and, based on recent experimental data, we will propose a view compatible with an emergent conceptual symmetry between the two apoptogenic extrinsic and intrinsic pathways. Elements of symmetry present in both the apoptogenic signalling pathways include: early activation of initiator caspases (feed-forwarded by a direct or post-mitocondrial effector caspase-mediated amplification loop in some cell types) and mitochondrial membrane permeabilization with required release of antagonists of active caspase inhibitors (IAPs) in high-level IAPs-expressing cells and apoptosome-mediated amplification of the caspase cascade more or less needed in different cell types. PMID- 14739600 TI - Degradation of poly(A)-binding protein in apoptotic cells and linkage to translation regulation. AB - We have recently shown that poly(A)-binding protein (PABP) is cleaved during poliovirus and Coxsackievirus infection by viral 3Cprotease and that 3Cprotease modification of a subset of PABP can result in significant translation inhibition. During apoptosis, translation undergoes significant down-regulation that correlates with caspase-3 mediated cleavage of several translation factors, including eIF4G, 4EBP1 and eIF2alpha. The fate of PABP in apoptotic cells has not yet been examined. Here we show that PABP levels decline significantly via proteolytic degradation in apoptotic HeLa, Jurkat and MCF7 cells. The degradation of PABP correlated with translation inhibition but lagged behind cleavage of eIF4GI. In apoptotic MCF7 cells translation inhibition occurred without modification of most translation factors and correlated with PABP degradation. PABP was not cleaved during incubation with several caspases, yet caspase 3 induced weak PABP degradative activity in cells lysates. Both the caspase inhibitor zVAD and calpain inhibitors blocked PABP cleavage in vivo, while the proteosome inhibitor MG132 induced PABP degradation. Protease(s) activated during apoptosis preferentially degraded PABP associated with ribosomes and translation factors, but not PABP in other cellular compartments. The data suggest that targeted degradation of PABP contributes to translation inhibition in apoptotic cells. PMID- 14739602 TI - The Siva-1 putative amphipathic helical region (SAH) is sufficient to bind to BCL XL and sensitize cells to UV radiation induced apoptosis. AB - The human Siva gene is localized to chromosome 14q32-33 and gives rise to the full-length predominant form, Siva-1 and a minor alternate form, Siva-2 that appears to lack the proapoptotic properties of Siva-1. Our recent work has shown that the missing region in Siva-2 encodes a unique twenty amino acid putative amphipathic helical region (SAH, residues 36-55 in Siva-1). Despite the fact that Siva-1 does not belong to the BCL-2 family, it specifically interacts with the anti-apoptotic protein BCL-XL and sensitizes MCF7 breast cancer cells expressing BCL-XL to UV radiation induced apoptosis. Deletion mutagenesis has mapped the necessary region to the SAH in Siva-1. In this paper we demonstrate that the SAH region in Siva-1 is sufficient to specifically interact with the anti-apoptotic members of the BCL2 family such as BCL-XL and BCL-2 but not its apoptotic member BAX. Using transient transfections and direct microinjection of synthetic SAH peptides, we also demonstrate that the SAH region is sufficient to inhibit the BCL-XL mediated cell survival and render MDA-MB-231 and MCF7 breast cancer cells expressing BCL-XL highly susceptible to UV radiation induced apoptosis. The underlying mechanism of action of SAH mediated inhibition of BCL-XL (and/or BCL2) cell survival appears to be due to loss of mitochondrial integrity as reflected in enhanced cytochrome c release leading to the activation of caspase 9 and finally caspase 3. PMID- 14739603 TI - Differential control of eosinophil survival by glucocorticoids. AB - Glucocorticoids are effective drugs for eosinophil-related disorders, such as asthma and allergy. Previous studies have demonstrated that glucocorticoids increase eosinophil apoptosis and block the survival effect of submaximal concentrations of interleukin-5 (IL-5). We investigated the effect of glucocorticoids on eosinophil survival in the presence of a higher concentration of IL-5 (1 ng/ml), comparable to IL-5 levels in bronchoalveolar lavage and sputum specimens from patients with asthma. In contrast to incubation in the presence of submaximal concentrations of IL-5, the addition of dexamethasone (DEX) to media containing 1 ng/ml IL-5 led to a significant increase in eosinophil cell viability from 58 +/- 6.9% to 87 +/- 2.4% ( p < 0.005) after 72 hours in culture. We found that RU486 blocked the DEX effect on cell viability confirming that glucocorticoid receptor functions are required. We investigated the possibility that the glucocorticoid enhancement of eosinophil survival may be due to an effect on IL-5 receptor expression. Our results show that the IL-5 associated decrease in IL-5 receptor alpha-subunit expression was blocked significantly after 24 hrs in culture with media containing IL-5 plus DEX compared to IL-5 alone. It is tempting to speculate that the observed glucocorticoid enhancement of eosinophil survival in the presence of elevated concentrations of IL-5 could be a mechanism that contributes to glucocorticoid resistance in asthma. PMID- 14739604 TI - Induction of necrotic tumor cell death by TRAIL/Apo-2L. AB - A great deal of enthusiasm is being generated for TRAIL (TNF-related apoptosis inducing ligand)/Apo-2L as a tumor therapeutic agent because it is cytotoxic to a variety of tumor cell types but not normal cells. Moreover, it is well documented that TRAIL/Apo-2L-induced tumor cell death is a caspase-dependent apoptotic process. Through the use of a transfected cell line expressing murine TRAIL/Apo 2L and a recombinant adenovirus encoding the murine TRAIL/Apo-2L cDNA (Ad5 mTRAIL) against two murine tumor cell lines [TRAMP-C2 (prostate adenocarcinoma) and Renca (renal adenocarcinoma)], we found that mTRAIL/Apo-2L also can kill tumor cells by inducing necrosis. Specifically, we observed the default method of mTRAIL/Apo-2L-induced death in TRAMP-C2 cells was via a necrotic process, characterized by the complete lack of an annexin V(+)/PI(-) population, SAPK/JNK phosphorylation, caspase activation, Bid cleavage, or cytochrome c release. Moreover, the inclusion of zVAD-fmk, an inhibitor of caspase activation, markedly enhanced mTRAIL/Apo-2L-mediated killing of TRAMP-C2. In contrast, apoptosis was induced in TRAMP-C2 using TNF, as measured by the criteria listed above, as was Renca by mTRAIL/Apo-2L. These results demonstrate the natural occurrence of both TRAIL/Apo-2L-induced apoptotic and necrotic signaling mechanisms within tumor cells. PMID- 14739605 TI - Potential mechanisms of leukemia cell resistance to TRAIL-induced apopotosis. AB - There are many factors contributing to the resistance to TRAIL (Tumor necrosis factor-related apoptosis-inducing ligand)-induced apoptosis. However, it is not clear whether the mechanism of resistance to TRAIL is constitutive or inductive. Therefore, the purpose of this study was to investigate the resistant mechanisms to TRAIL at different levels in the apoptotic pathway. The human T-lymphoblastic leukemic CEM cell line showed more resistant to TRAIL-induced apoptosis compared with the human chronic myeloid leukemic K562 cell line. Lower level of constitutive caspase-8 expression in the CEM cell line led to a poor response to both TRAIL-induced activation of caspase-3 and reduction in the mitochondrial membrane potential (DeltaPsim). There was no significant difference in the constitutive levels of NF-kappaB in CEM and K562 cell lines. However, CEM cells showed a faster response to TRAIL-induced NF-kappaB activation than K562 cells. TRAIL-induced regulation of Bcl-2 family of proteins included an up-regulation in Bcl-2/Bcl-XL and a down-regulation in Bax. IAPs, such as XIAP, cIAP-1, cIAP-2 and Survivin were all up-regulated during the treatment with TRAIL. In summary, our data suggest that the leukemic cells resistance to TRAIL-induced apoptosis might be due to the deficiency in the constitutive caspase-8 expression. Development of potential resistance to apoptosis by TRAIL can occur in both TRAIL-resistant and TRAIL-sensitive leukemic cells. PMID- 14739606 TI - Early events in Bcl-2-enhanced apoptosis. AB - Transfection of PC12 pheochromocytoma cells with bcl-2 potentiates apoptosis induced by the antimitotic agent, neocarzinostatin (NCS). The mechanism of potentiation involves caspase 3-dependent cleavage of Bcl-2 to its pro-apoptotic counterpart, but the cellular events proximal to caspase 3 activation in this system are not known. Two min after initiation of NCS treatment, Bax begins to translocate from cytosol to the mitochondria; the mitochondrial localization of Bax persists for 30 min after NCS treatment. At the same time, cytochrome C is released from the mitochondria to cytosol. The mitochondrial membrane potential exhibits differential change in mock- and bcl-2 -transfected PC12 cells. In mock transfected PC12 cells, the mitochondrial membrane potential increases immediately, peaks at 15 min following initiation of NCS treatment, and drops thereafter. In contrast, in bcl-2 -transfected PC12 cells, the membrane potential drops immediately following NCS treatment. Caspase 9 is activated and peaks at 10 min in both mock- and bcl-2 transfected PC12 cells, however, the peak activity of caspase 9 is higher and caspase 9 activation lasts longer (30 min) after the treatment in bcl-2 transfectants. Not until 30 min after initiation of a 1 h treatment with NCS is Bcl-2 protein cleaved in bcl-2 -transfected cells. Thus, in bcl-2 -transfected cells, the mitochondrial membrane potential drops and cytochrome C is released from the mitochondria despite the presence of large amounts of intact mitochondrial Bcl-2. This makes it unlikely that cleavage of Bcl-2 is the only factor involved in potentiation of NCS-induced apoptosis by Bcl 2. PMID- 14739607 TI - Apoptosis modulatory activities of transiently expressed Bcl-2: roles in cytochrome C release and Bax regulation. AB - Bcl-2 and Bcl-X(L) are pro-survival members of the Bcl-2 family. These proteins have been shown to antagonize the pro-apoptotic activity of Bax and promote cell survival through blocking Bax translocation from the cytosol to mitochondria and by preventing the release of cytochrome c. However, it has been recently reported that transiently expressed Bcl-2 unexpectedly leads to significant cell toxicity. To study this intriguing phenomenon, we have carried out further analyses into the properties of transiently expressed Bcl-2. We found that various isoforms of human and different species of Bcl-2 were equally capable of inducing apoptosis. In addition, we discovered that transient expression of Bcl-2, unlike its pro survival homolog Bcl-X(L), can lead to the release of cytochrome c from mitochondria and that the resulting cell death can be inhibited by caspase and calpain inhibitors. Moreover, we have shown that unlike the pro-apoptotic protein Bid, the toxicity associated with the transient expression of Bcl-2 occurs independent of the activity of the endogenous Bax. Finally, we found that in spite of its intrinsic toxicity, transiently expressed Bcl-2 is fully capable of blocking the ectopically expressed Bax from localizing to mitochondria. Taken together, these studies demonstrate that transiently expressed Bcl-2 displays opposing functional properties. PMID- 14739608 TI - MTS-conjugated-antiactive caspase 3 antibodies inhibit actinomycin D-induced apoptosis. AB - Caspase 3 is critically involved in the pathway of apoptosis. We have conjugated a MTS-transport-peptide to monoclonal and polyclonal anti-caspase-3 antibodies to suppress Actinomycin D-induced apoptosis in human lymphoma T cells. The advantage of using trans-membrane antibodies compared to conventional apoptosis inhibitors is their specific target recognition in the living cell and their lower toxicity compared to conventional apoptosis inhibitors. We could show that a MTS-transport peptide modified monoclonal anti-caspase-3 antibody reduces Actinomycin D induced apoptosis, as shown by DNA ladder electrophoresis and cell death ELISA. These results indicate that antibodies have a therapeutic potential to inhibit apoptosis in a variety of diseases. PMID- 14739609 TI - Camptothecin-induced apoptosis in non-small cell lung cancer is independent of cyclooxygenase expression. AB - Recent observations show a positive correlation between the expression of cyclooxygenase (COX), especially COX-2), and cancer development. Here we tested the hypothesis that expression of COX-2 could influence apoptosis in lung cancer cell lines. To address this question, we determined the effects of camptothecin induced apoptosis on three lung cancer cell lines which over express COX-1 (CORL23), COX-2 (MOR-P) and neither isoform (H-460), and determine if these effects were prostaglandin mediated. We also compared the effects of non selective and isoenzyme selective COX-2 inhibitors on camptothecin-induced apoptosis in these three cell lines. Camptothecin induced apoptosis in all three cell lines independently of COX-1 or COX-2 expression. Indomethacin, a non selective COX inhibitor and NS398, a selective COX-2 inhibitor had no effect on camptothecin-induced apoptosis at concentrations that abolished prostaglandin production. In conclusion, these finding suggest that the COX pathway is not involved in camptothecin-induced apoptosis of non-small cell lung cancer cell lines. PMID- 14739610 TI - Cox-2 is needed but not sufficient for apoptosis induced by Cox-2 selective inhibitors in colon cancer cells. AB - The role of Cox-2 in NSAID-induced apoptosis is debated. We studied the role of Cox-2 inhibition in apoptosis induced by a selective Cox-2 inhibitor, SC236 (a structural analogue of celecoxib) in two colon cancer cell lines, HT29 (expressing Cox-2 protein) and HCT116 (not expressing Cox-2 protein). Apoptosis was quantified by flow cytometry. SC236 0-75 microM decreased cell numbers and induced apoptosis to identical levels in HT29 and HCT116 cells. However, SC236, concentrations >75 microM reduced Cox-2 protein expression in HT29 cells and induced greater levels of apoptosis in HT29 than in HCT116 cells. In contrast, sulindac sulfide (SSD) (which inhibits Cox-1 and Cox-2) 0-200 microM or sulindac sulfone (SSN) 0-500 microM (without significant activity against Cox-1 or Cox-2) caused identical decreases in cell number and increases in apoptosis in HT29 and HCT116 cells. Neither SSD nor SSN altered the expression of Cox-2 in HT29 cells. To determine that the higher levels of apoptosis in HT29 cells with SC236 >75 microM were related to decreased Cox-2 protein levels, we decreased Cox-2 protein expression in HT29 cells with curcumin (diferuloylmethane) and studied its effect on SC236-induced apoptosis. Curcumin augmented apoptosis induced by SC236 in HT29 cells but not in Cox-2 lacking HCT116 cells. In conclusion, selective Cox-2 inhibitors can induce apoptosis independent of Cox-2 expression. However they may selectively target cells that express Cox-2 by decreasing their Cox-2 protein expression. PMID- 14739611 TI - Apoptosis participates to liver damage in HSV-induced fulminant hepatitis. AB - BACKGROUND: HSV fulminant hepatitis is a rare pathology. Rapid hepatic failure, as a consequence of extended liver damage, has generally been attributed to necrosis. As apoptosis can constitute another way for hepatocytes to die, we decided to investigate whether programmed cell death took place during HSV fulminant hepatitis. METHODS: Liver sections were obtained from two cases of fulminant herpetic hepatitis as well as from hepatitis B virus and Rickettsia infected livers. Herpes simplex virus infection was confirmed using in situ hybridization. Apoptosis was assessed by histopathological examination, p53, activated-caspase 3 and Fas immunohistochemistry and TUNEL labeling. RESULTS: We report that the number of cells expressing activated-caspase 3 was largely increased in fulminant herpes simplex virus hepatitis, when compared to livers chronically infected by hepatitis B virus or from a Rickettsial acute hepatitis. Apoptosis of hepatocytes was confirmed by a positive double-staining for activated-caspase 3 and hepatocytes. Finally, the apoptotic process has progressed beyond the step of nuclear DNA cleavage as demonstrated by TUNEL labeling. CONCLUSION: These data as a whole show that apoptosis is responsible, at least partially, for liver damage during HSV fulminant hepatitis. PMID- 14739612 TI - Apoptosis does not affect the vasculature of the corpus luteum of pregnancy in the rat. AB - There has been much research into the mechanics of angiogenesis and many studies have demonstrated that newly formed vessels regress during angiogenesis. This vascular involution has been shown to involve basement membrane dissolution and endothelial cell apoptosis. The corpus luteum provides an ideal in vivo model to study physiologic angiogenesis and studies have shown that involution of newly formed vessels occurs during corpus luteum regression. However, few studies to date have investigated the role of apoptosis on the vasculature which develops during pregnancy. By the use of the TUNEL technique to detect apoptotic cells and immunohistochemistry to distinguish between endothelial cells and pericytes, this present study demonstrated that the vasculature of the corpus luteum of pregnancy in the rat does not undergo apoptosis. PMID- 14739613 TI - Tissue transglutaminase mRNA expression in apoptotic cell death. AB - INTRODUCTION: Tissue transglutaminase (t.TG) is an enzyme that catalyzes the cross-linking of intracellular proteins, thus assembling a protein scaffold that prevents leakage of intracellular components. t.TG is activated during the apoptotic cell death cascade and plays a key role in the formation of apoptotic bodies. The aim of this study was to determine to what amount t.TG-mRNA becomes expressed during apoptosis and whether the t.TG-mRNA expression level could be used as trace marker of recent apoptosis and in individual cases for quantification of apoptosis. METHODS: Expression of t.TG-mRNA was determined using TaqMan based, real-time RT-PCR, a semi-quantitative RT-PCR technique. The t.TG-mRNA expression was measured in cultured cells (MCF-7, human endothelial cells) and in peripheral blood mononuclear cells (PBMCs) before and after induction of apoptosis in vitro. RESULTS: The TaqMan RT-PCR of t.TG proved to be reliable, reproducible (CV's inter and intraassay precisions of 0.8-2.8%, measured at two levels), and specific for apoptotic cell death. t.TG-mRNA expression increases in response to apoptosis induction and is not expressed during the process of necrotic cell death. The expression during apoptotic cell death changes in the dose dependent manner in cultured cells as well as in the PBMCs, treated in vitro. The increase t.TG-mRNA expression level was up to 20 times, depending on the intensity of the apoptosis induction treatment and incubation time afterwards. PBMCs of patients with myelodysplasia showed spontaneous expression of t.TG-mRNA in agreement with their increased apoptotic cell death in vivo. CONCLUSION: t.TG-mRNA expression increases significantly in response to apoptosis inducing treatment. The observed changes are dose and time dependent. This leads to the conclusion that t.TG expression can be used as a trace marker for detection and quantification of apoptosis. PMID- 14739614 TI - Caspase-9 activation in hypoxic human corneal epithelial cells. AB - The purpose of this study was to determine the effect of hypoxia on caspase-8 and -9 gene and protein expression and activity in corneal epithelium. Non transformed human corneal epithelial cells (HCEC) were cultured in 2% oxygen. A cDNA expression array coupled with densitometric analysis was used to compare relative mRNA expression levels of 96 apoptosis-related genes in hypoxic and normoxic HCEC. Caspase-8, caspase-9, FLIP, Fas, FasL, and TNFalpha protein expression was assessed further using Western blot analysis and ELISA. Caspase-8 and -9 activities were measured using a fluorometric activity assay. Hypoxia did not affect caspase-8 or -9 gene or protein expression in HCEC, however caspase-9 activity was significantly increased. Hypoxia significantly suppressed the activity of caspase-8. FLIP and Fas gene and protein expression were not significantly altered in hypoxic cells compared to normoxic controls. mRNA and protein levels of TNFalpha and TNFR-1 were significantly decreased, while FasL mRNA and proteins levels were significantly increased in hypoxic HCEC. In corneal epithelium stressed by hypoxia caspase-9 activity is upregulated, suggesting that apoptosis proceeds via the mitochondrial pathway. Caspase-8 activity may be suppressed because the loss of TNFalpha and TNFR-1 gene and protein expression inhibits the initial formation of a death signaling complex. PMID- 14739615 TI - Discovery and development of anti-angiogenic peptides: A structural link. AB - Cancer is a disease promoted by excess angiogenesis. Interference with this process poses an attractive approach to controlling aberrant tumor growth, a hypothesis first proposed in the early 1970s that led to world-wide focus on identifying and developing angiogenesis inhibitors, which currently number in the hundreds. This review surveys the discovery and development of anti-angiogenic protein fragments and peptides, with a slant towards understanding their structure-function relationships to aid in the design of better therapeutic agents. PMID- 14739616 TI - Generation and use of a tailored gene array to investigate vascular biology. AB - Vasculogenesis, angiogenesis and vascular remodelling are complex processes where the fate of several cell types is determined by different signalling networks. Many of these networks ultimately function by changing the abundance of RNA transcripts within the cells which constitute blood vessel walls. Researchers can now map these transcript abundance changes using gene array technology. In this review, we describe the design, production and use of a gene array specifically tailored to investigate vascular biology. We describe the advantages of tailored gene arrays, and give detailed protocols based on our experience to allow the reader to use such gene arrays to generate meaningful data. We list the issues to consider when choosing and verifying the genes and splice variants included in an array, and describe our use of Arabidopsis sp. RNA spikes for quality control. We present data that illustrates the absolute necessity for both technical and biological replicates to be incorporated in the design of gene array experiments using primary cells such as HUVECS. Finally, we describe methods for the normalisation and interpretation of the data that gene arrays produce. The approach to gene array technology described here is easily within reach of the budget and expertise of most academic research groups. PMID- 14739617 TI - alphav beta 3 and alphav beta 5 integrin antagonists inhibit angiogenesis in vitro. AB - Although angiogenesis is believed to require cell-extracellular matrix interactions which are mediated in part via integrins alphav beta 3 and alphav beta 5, a formal demonstration that alphav beta 3 and alphav beta 5 are involved in endothelial-cell invasion and capillary-like tube formation is still required. This has arisen from the cellular complexities which occur in vivo and the difficulty in finding appropriate in vitro model systems. Here we have used a three-dimensional assay which employs bovine aortic and microvascular endothelial cells, to show that alphav beta 3 and alphav beta 5 regulate angiogenesis in vitro. We cloned and characterized 350-450 bp regions of the bovine homologues of alphav, beta 3 and beta 5, covering much of the beta -propeller and A-domain regions, and show that they are >95% identical to their human orthologues. We used cyclic peptides EMD 121974, 85189 and 66203, which selectively inhibit alphav beta 3 and alphav beta 5, but not gpIIbIIIa or alpha5 beta 1, to probe in vitro angiogenesis induced by angiogenic cytokines in three-dimensional fibrin or collagen gels. We found that these peptides are potent inhibitors of endothelial cell invasion and differentiation induced by vascular endothelial growth factor-A or fibroblast growth factor-2 but do not affect the unstimulated cells in 3D culture. Inhibition was greatest when cells were grown on fibrin, but also occurred on collagen I which is not a recognized ligand for alphav beta 3. These findings demonstrate the requirement for endothelial cell alphav beta 3 and alphav beta 5 integrins during angiogenesis in vitro, and are in accord with the proposed therapeutic application of alphav beta 3 and alphav beta 5 antagonists. PMID- 14739618 TI - Preliminary studies on the anti-angiogenic potential of pomegranate fractions in vitro and in vivo. AB - We previously showed pomegranate seed oil and fermented juice polyphenols to retard oxidation and prostaglandin synthesis, to inhibit breast cancer cell proliferation and invasion, and to promote breast cancer cell apoptosis. Here we evaluated the anti-angiogenic potential of these materials in several ways. We checked a possible effect on angiogenic regulation by measuring vascular endothelial growth factor (VEGF), interleukin-4 (IL-4) and migration inhibitory factor (MIF) in the conditioned media of estrogen sensitive (MCF-7) or estrogen resistant (MDA-MB-231) human breast cancer cells, or immortalized normal human breast epithelial cells (MCF-10A), grown in the presence or absence of pomegranate seed oil (SESCO) or fermented juice polyphenols (W). VEGF was strongly downregulated in MCF-10A and MCF-7, and MIF upregulated in MDA-MB-231, overall showing significant potential for downregulation of angiogenesis by pomegranate fractions. An anti-proliferative effect on angiogenic cells was shown in human umbilical vein endothelial cell (HUVEC) and in myometrial and amniotic fluid fibroblasts, and inhibition of HUVEC tubule formation demonstrated in an in vitro model employing glass carrier beads. Finally, we showed a significant decrease in new blood vessel formation using the chicken chorioallantoic membrane (CAM) model in vivo. 'In sum, these varied studies employing different models in different laboratories overall demonstrate for the first time an anti-angiogenic potential of pomegranate fractions, suggesting further in vivo and clinical investigations (for updates: info@rimonest.com). PMID- 14739619 TI - Verotoxin sensitivity of ECV304 cells in vitro and in vivo in a xenograft tumour model: VT1 as a tumour neovascular marker. AB - Binding of verotoxin 1 (VT1) via its receptor (globotriaosylceramide/Gb3) to both endothelial and tumour cell subsets suggests that VT1 may have both antineoplastic and antiangiogenic potential. We investigated this potential using the ECV304 cell line, which, although identified as a bladder carcinoma cell line, displays some endothelial characteristics, including tubule formation (differentiation) following appropriate stimulation. Differentiated ECV304 cells retained Gb3 expression/VT1 sensitivity. VT1 internalization and retrograde transport through the Golgi to the ER was observed. ECV304 xenografts in immunocompromised mice were invasive but surprisingly poorly vascularized. Intratumoural VT1 injection significantly reduced ECV304 xenograft growth and enhanced mouse survival. Gb3 expression was decreased in residual tumour, likely due to cell cycle arrest. Untreated ECV304 xenograft sections bound VT1 throughout the tumour. Anti-von Willebrand factor (vWF) antibody staining for neovasculature showed only morphologically atypical, indistinct structures, unlabelled by VT1. Human bladder carcinoma samples were, in contrast, highly vascular and blood vessels were 100% co-labelled by anti-vWF antibody and VT1, with no extravascular staining. These results suggest that ECV304 xenografts are not characteristic of bladder carcinoma in terms of Gb3 expression, and that VT1 staining may provide a new reliable index of tumour neovasculature. We conclude that ECV304 cells are not an appropriate in vivo model of either tumour angiogenesis or bladder carcinoma. These studies, nevertheless, further demonstrate the in vivo antineoplastic and antiangiogenic potential of VT1, and show that Gb3 is expressed in cells undergoing in vitro 'vascular' differentiation, and in the neovasculature of human bladder carcinomas. PMID- 14739620 TI - Inhibition of angiogenic initiation and disruption of newly established human vascular networks by juice from Morinda citrifolia (noni). AB - noni, the juice of the fruit from the Morinda citrifolia plant, has been used for centuries as a medicinal agent. We tested the effects of noni juice in a three dimensional fibrin clot matrix model using human placental vein and human breast tumor explants as sources for angiogenic vessel development. Noni in concentrations of 5% (vol/vol) or greater was highly effective in inhibiting the initiation of new vessel sprouts from placental vein explants, compared with initiation in control explants in media supplemented with an equivalent amount of saline. These concentrations of noni were also effective in reducing the growth rate and proliferation of newly developing capillary sprouts. When used at a concentration of 10% in growth media, noni was able to induce vessel degeneration and apoptosis in wells with established capillary networks within a few days of its application. We also found that 10% noni juice in media was an effective inhibitor of capillary initiation in explants from human breast tumors. In tumor explants which did show capillary sprouting, the vessels rapidly degenerated (2-3 days) in those exposed to media supplemented with 10% noni. PMID- 14739621 TI - Induction of focal angiogenesis through adenoviral vector mediated vascular endothelial cell growth factor gene transfer in the mature mouse brain. AB - Vascular endothelial growth factor (VEGF) is a potent endothelial cell mitogen and morphogen, which stimulates angiogenesis in a wide variety of tissues and lesions in vivo. In this study, we applied adenoviral vector delivered human VEGF165 cDNA to develop focal non-tumor angiogenesis in the mature mouse brain. Seventy-two adult CD-1 mice underwent Ad h VEGF, Ad lacZ, and saline injection for up to fourweeks. An adenoviral suspension containing 1 x 10(9) particles was injected stereotactically into the right hemisphere of the brain. The results showed that VEGF expression was increased in the Ad h VEGF transduced mice compared to Ad lacZ or saline injected mice ( P < 0.05). VEGF-positive cells were mainly located in the injection hemisphere of Ad h VEGF transduced mice. Quantitative vessel counting showed that microvessels in the Ad h VEGF transduced mice increased following 2 weeks of Ad h VEGF gene transfer compared to the other two groups (Ad h VEGF:241 +/- 19 vs. Ad lacZ :148 +/- 17 and Saline:150 +/- 14 vessels/mm2, P < 0.05). Morphology showed typical angiogenic changes. PCNA positive staining confirmed these microvessels were actively proliferating. Our study demonstrates that Ad h VEGF-induced VEGF hyper-stimulation causes focal angiogenesis in the mature mouse brain. This novel method of inducing in vivo brain focal angiogenesis provides an opportunity to study the molecular mechanisms independent of the confounding effects of upstream inciting stimuli such as ischemia or tumor. PMID- 14739622 TI - Observer variability in ECG interpretation for thrombolysis eligibility: experience and context matter. AB - BACKGROUND: Despite the known benefit of thrombolysis it remains underutilized among eligible patients with acute myocardial infarction. We sought to determine whether potential errors in ECG interpretation might be a contributing factor and to what extent clinical history, a checklist outlining recognized inclusion criteria and a computerized interpretation would influence reliability and accuracy. METHODS: Seventy-five ECGs were interpreted on 8 separate occasions by 9 clinicians (3 cardiologists, 3 cardiology fellows, 3 medical residents) according to a 2 x 2 x 2 factorial design. RESULTS: The overall level of agreement among all raters was substantial with a kappa (kappa) of 70.4%. Intra observer ECG reading reliability was stronger among cardiologists (CC) as compared with cardiology fellows (CF) and medical residents (MR). Similarly, inter-observer reliability was substantial to very good and a gradient was seen with greater reliability among CC, followed by CF, then MR ( P = 0.0013). CC recommended thrombolysis significantly more frequently ( p < 0.001) than either CF or MR. Trainees were biased by the presence of a computerized ECG interpretation resulting in a decision to recommend thrombolysis administration less often. CONCLUSION: The reliability of ECG interpretation for deciding to administer thrombolysis was substantial; there was a gradient from lowest to highest commensurate with training and experience. Errors in thrombolysis eligibility are influenced by clinical history and the presence of a computerized ECG interpretation among less experienced clinicians. PMID- 14739623 TI - Acquired von Willebrand syndrome: its pathophysiology, laboratory features and management. AB - Acquired von Willebrand syndrome is a bleeding disorder associated with various underlying diseases. The clinical manifestations are similar to congenital von Willebrand disease. Diagnosis is mainly confirmed by decrease of ristocetin cofactor activity (vWF:RCo) and collagen binding activity (vWF:CBA) and by vWF multimeric analysis usually with selective loss of large multimers. Various pathogenic mechanisms have been proposed, including development of autoantibodies to von Willebrand factor (vWF), adsorption of vWF onto tumor cells or activated platelets, increase of vWF proteolysis, and mechanical destruction of vWF under high shear stress. Following the treatment of underlying disorders, desmopressin (DDAVP) is a first intention therapeutic option. Factor VIII/vWF concentrates and high dose immunoglobulin infusions are reserved for patients unresponsive to DDAVP. PMID- 14739625 TI - Synergism of aspirin and heparin with a low-frequency non-invasive ultrasound system for augmentation of in-vitro clot lysis. AB - BACKGROUND: Aspirin, glycoprotein IIb/IIIa inhibitors and heparin are routinely used in acute coronary syndromes. Previously we showed that there is synergism between ultrasound and heparin and tirofiban in augmenting blood clot disruption. However, there is a little data on a possible synergism of low-frequency ultrasound with aspirin for in-vitro clot dissolution, and especially on the combination of aspirin with heparin and/or glycoprotein IIb/IIIa inhibitors. MATERIALS AND METHODS: Human blood clots (n = 320) were incubated for 10 or 20 minutes in saline containing aspirin alone or combined with heparin and/or tirofiban and/or eptifibatide. Clots were randomly treated with low-frequency ultrasound (27.3 kHz) or incubation only. The percent clot weight loss and the incremental effect of ultrasound were calculated. RESULTS: The most significant incremental effect of ultrasound on clot weight reduction was detected with aspirin alone (5.2 +/- 2.3% and 5.2 +/- 2.6% after 10' and 20', p = 0.04 and p = 0.06, respectively) and in combination with heparin (8.8 +/- 2.5% and 11.5 +/- 2.7% after 10' and 20', p = 0.01 and p = 0.0001, respectively). The greatest absolute magnitude of clot weight reduction was observed with ultrasound combined with aspirin and heparin (48.5 +/- 9.5% after 20'). The addition of tirofiban or eptifibatide to aspirin, heparin and ultrasound did not increase clot lysis. However, eptifibatide had significantly better synergism than tirofiban (p = 0.025 and p = 0.015, after 10 and 20 minutes, respectively). CONCLUSIONS: Aspirin alone or in combination with heparin results in significant augmentation of clot lysis and is synergistic with application of low-frequency ultrasound for 10 and 20 minutes only. These results may have important implications for a possible use of low-frequency ultrasound in treatment algorithms of acute coronary syndromes. PMID- 14739624 TI - Thrombin domains: structure, function and interaction with platelet receptors. AB - Thrombin plays a pivotal role in different biological phenomena, such as hemostasis, thrombosis, and cell differentiation. Indeed this protease catalyzes the conversion of fibrinogen into fibrin, the activation of coagulation factors V, VIII, XI, and XIII, but is also involved in the activation of many cell types and platelets. Thrombin bears some recognition domains and insertion loops, not found among other serine proteases of the coagulation system. In this review the properties of these thrombin domains, which regulate the specificity of the enzyme's interaction with substrates and inhibitors, are particularly emphasized. The example of thrombin interaction with the platelet membrane receptors, namely GpIb and PAR1, shows how the concerted action of the insertion loops and recognition domains is the key to solve the apparent enigma as to how thrombin can be at the same time a very efficient and specific enzyme for different substrates and inhibitors. PMID- 14739626 TI - Efficacy and safety of a new streptokinase regimen with enoxaparin in acute myocardial infarction. AB - OBJECTIVE: To compare a new streptokinase regimen combined with either enoxaparin or unfractionated heparin (UFH) and the traditional streptokinase regimen combined with UFH in patients with acute myocardial infarction (AMI). METHODS: 412 patients (<75 years), hospitalized within 6 hours of the onset of chest pain, were allocated thrombolytic therapy by the treating physician: streptokinase 0.75 MU/10 minutes, repeated if no coronary reperfusion after one dose, plus enoxaparin 40 mg intravenously followed by 1 mg/kg bodyweight subcutaneously at 12-hour intervals for 5-7 days (n = 102); the same streptokinase regimen plus UFH 1000 IU/60 minutes intravenously for 48-72 hours ( n = 106); or streptokinase 1.5 MU/60 minutes plus the same UFH regimen (n = 204). All patients received 250-325 mg aspirin/day. Coronary reperfusion rates, 30-day mortality and hemorrhagic complications were recorded. RESULTS: Coronary reperfusion rates with 0.75 streptokinase + enoxaparin (78.4%) and 0.75 streptokinase + UFH (74.5%) were significantly higher than those with 1.5 streptokinase + UFH (62.2%), but there was no significant difference between the groups receiving the new regimen. Overall 30-day mortality (6.3%) was significantly lower than with 1.5 streptokinase + UFH (12.7%) ( p = 0.037). The incidence of major and minor hemorrhagic events was similar in all groups. CONCLUSIONS: The accelerated streptokinase regimen was well tolerated and resulted in a significantly higher coronary reperfusion rate and significantly lower mortality compared with the traditional regimen. The 0.75 streptokinase + enoxaparin combination was at least as efficacious as the 0.75 streptokinase + UFH combination and is preferred because of its ease of administration and predictable anticoagulant effect. PMID- 14739627 TI - Pre-hospital fibrinolysis followed by angioplasty or primary angioplasty in acute myocardial infarction: the long-term clinical outcome. AB - BACKGROUND: Randomized trials comparing primary angioplasty and in-hospital fibrinolysis in acute myocardial infarction (AMI) have shown an advantage for primary angioplasty. The long-term follow-up of pre-hospital fibrinolysis followed by elective or rescue coronary angioplasty versus primary angioplasty is not well established after acute myocardial infarction. This study sought to assess the long-term clinical outcome of patients with AMI having either received pre-hospital fibrinolysis optimized by coronary angioplasty or primary angioplasty. METHODS: We conducted a retrospective analysis involving 318 patients who either underwent primary angioplasty ( n = 157) or received pre hospital fibrinolysis followed by an angioplasty (rescue or elective) ( n = 161) within 6 hours of the onset of chest pain. RESULTS: The groups were similar regarding their baseline characteristics except for the ages. No difference was noted for in-hospital mortality (primary PTCA group: 2.48%, combined group: 2.54%; p = ns) with no increased risk of hemorrhage. The 3-year mortality was not significantly different in the two groups (9.7% vs. 4.9%; p = 0.15). Regarding major adverse cardiac events (29.5% vs. 37.5%; p = 0.23), reintervention (22.5% vs. 23.2%; p = 0.99) or target lesion revascularization (16.1% vs. 14.7%; p = 0.68), the groups were statistically similar. CONCLUSION: These data from real life practice emphasize the safety and similar benefits on the long-term clinical outcome of AMI patients having undergone either pre-hospital fibrinolysis followed by angioplasty or primary angioplasty. PMID- 14739628 TI - Distance from the coronary ostium to the culprit lesion in acute ST-elevation myocardial infarction and its implications regarding the potential prevention of proximal plaque rupture. AB - BACKGROUND: Shorter distances from the coronary ostia to culprit lesions have been associated with a higher incidence of adverse outcomes in ST elevation acute myocardial infarction (STEMI). As drug-eluting stents are associated with low rates of restenosis and formation of a stable intima, we sought to develop a mathematical model to estimate how far down the coronary artery a drug-eluting stent would have to be placed to theoretically mitigate the risk of proximal plaque rupture. OBJECTIVES AND METHODS: Distances from the ostia to the end of the culprit lesions were planimetered in 1,914 patients from the TIMI 14, INTEGRITI, FASTER and ENTIRE/TIMI 23 trials. RESULTS: The first 60 mm of the coronary artery contained 75% of STEMI culprit lesions. The median distance from the vessel ostium to the end of the culprit lesion was 43 mm (mean 50 +/- 34) and the relative distance from the vessel ostium to the end of the lesion was 29% (mean 33 +/- 17%) of the total culprit artery length. Diabetes was the only baseline clinical characteristic associated with a longer absolute distance to the end of the culprit lesion (46 mm vs. 43 mm, p = 0.03) as well as relative to total artery length (31% vs. 29%, p = 0.04). Median distances from the artery ostium to the end of the culprit lesion were shortest among the left anterior descending culprits (40 mm), followed by circumflex lesions (43 mm) and then right coronary artery lesions (47 mm, 3-way p < 0.0001). CONCLUSION: The majority of culprit lesions in STEMI are contained within the proximal 30% of the major epicardial coronary arteries, but the distance varies depending upon which epicardial artery is involved. Cumulative distribution functions are presented to allow estimation of the percent of culprit lesions lying proximal to any given distance from the ostium to model the feasibility of prophylactic drug-eluting stenting to minimize the risk of subsequent proximal plaque rupture. PMID- 14739629 TI - Age and first INR after initiation of oral anticoagulant therapy with acenocoumarol predict the maintenance dosage. AB - BACKGROUND: This retrospective study was performed to develop a model to predict the maintenance dosage of the vitamin K antagonist acenocoumarol, based upon the first INR after a standard initial dosage regimen. MATERIAL AND METHODS: Outpatients with atrial fibrillation ( n = 284) and initial regimens of 6-4 or 6 4-2 mg acenocoumarol on day 1, 2 and 3, respectively, were included. The maintenance dosage of the period 3-6 months after the installment was related to the first INR after those standard initial dosage regimens, because in that period the INR was 76% of the time within the therapeutic range and therefore considered suitable to perform the analysis. RESULTS: A clear relation was found between the first INR, the maintenance dosage and the age. A model that predicts the maintenance dosage immediately after the standard initial dosage and based on the first INR and adjusted for age, has been developed, according to the formula: required dosage = 5.03-1.65 * ln (first INR) - 0.01 * age. CONCLUSION: We have developed a formula to predict the maintenance dosage of acenocoumarol. With this formula it is possible to install this maintenance dosage and thus achieve oral anticoagulant therapy within the therapeutic range at an earlier stage. This will have to be shown in a prospective study. PMID- 14739630 TI - Polymorphism induced sensitivity to warfarin: a review of the literature. AB - Warfarin is a widely prescribed anticoagulant used for prophylaxis and treatment of venous and arterial thrombosis. Although warfarin is considered very efficacious, it has substantial risks associated with its use, specifically the risk of hemorrhage. Genetic variants associated with the metabolism of (S) warfarin by cytochrome P450 2C9 may have specific implications on untoward effects. Twelve CYP2C9 allelic variants have been identified, of which CYP2C9*3 and CYP2C9*2 are the most clinically important. Studies have demonstrated that initial dosing of warfarin with CYP2C9*3 with a five-milligram dose caused an increase in the international normalized ratio and significant risk of bleeding. Studies conducted with CYP2C9*2, on the other hand are conflicting. Some data suggest that the CYP2C9*2 variant is associated with an increased propensity for bleeding whereas other studies do not demonstrate a substantial risk of adverse events. Researchers suggest that detection of genetic variants in susceptible individuals will not only decrease the risks associated with warfarin therapy but also decrease costs of adverse events. PMID- 14739631 TI - Quality assessment of anticoagulation dose management: comparative evaluation of measures of time-in-therapeutic range. AB - BACKGROUND: The results of clinical trials often hinge on the quality of oral anticoagulation management, yet the quality of such management is frequently not mentioned or measured. Time-in-therapeutic range (TTR) is one measure of quality of anticoagulation dose management, but various methodologies exist for measuring TTR. OBJECTIVE: This study was initiated to compare three commonly used methodologies for measuring TTR to see how they compare within the same cohort of patients. PATIENTS/METHODS: Three common methodologies of calculating time-in therapeutic range were analyzed retrospectively in a cohort of patients over six two-month intervals. Additional analysis was performed for three and six-month intervals. The methodologies included fraction of INRs in range; cross-section of the files; and linear interpolation. RESULTS: Fraction of the INRs in range and cross-section of the files methodologies gave similar results, while linear interpolation yielded significantly shorter time-in-range for the two-month, three-month, and six-month intervals measured. The advantages and disadvantages of each methodology are discussed. CONCLUSIONS: The decision of which method to use should be based on clinic size, information desired, and clinic resources for ease of applying either of the methods in clinical practice. Each of these methodologies has their limitations and the question remains as to which method best reflects the quality of anticoagulation management. Regardless of these limitations, investigators are urged to employ at least one method of measuring the quality of oral anticoagulation management so as to better assess the validity of the clinical outcomes. PMID- 14739632 TI - Effects of angiotensin converting enzyme inhibitors on thrombotic mediators: potential clinical implications. AB - Angiotensin-converting enzyme inhibitors (ACE-I) were initially developed as therapeutic agents targeted for the treatment of hypertension. Since the initial application of these agents, several additional clinical indications have been identified such as coronary artery disease, stroke, congestive heart failure and prevention of diabetes-related complications. In addition to the blood pressure lowering effects, this class of agents has the ability to restore endothelial function, decrease oxidative stress and enhance endogenous fibrinolysis. Moreover, ACE-I possesses antiplatelet effects as well as antiproliferatory and antimigratory effects on smooth muscle cells. This article links the effects of ACE-I on thrombotic mediators to the potential clinical implications in the setting of coronary artery disease. PMID- 14739633 TI - The clinical thrombosis center and clinical thrombologist: a new US health systems paradigm for the management of venous thromboembolic disease. AB - New paradigms for the diagnosis, prophylaxis, acute treatment, and ongoing management of patients with venous thromboembolic disease (VTE), a better understanding of the genotypic and phenotypic mechanisms of thrombophilic states, and the possibility of a greatly expanded armamentarium of antithrombotic therapies are necessitating a more formalized and systematic approach to VTE management. This has required many US healthcare institutions to develop piecemeal approaches in management models for VTE utilizing local champions from a variety of subspecialties. Development of a formalized Clinical Thrombosis Center from an already established Anticoagulant Management Service utilizing a clinical thrombologist, a new role for a physician who has developed expertise in anticoagulation and VTE management, presents a new paradigm in which this disease may be approached at a formalized, institutional level. Thus the clinical thrombologist working through a Clinical Thrombosis Center can develop a system of-care approach to link the rapid advances in the field of thromboembolism to clinical applications, formulate evidence-based disease management guidelines, and conduct patient-oriented translational clinical research in VTE. PMID- 14739635 TI - BACUS: A Bayesian protocol for the identification of protein NOESY spectra via unassigned spin systems. AB - NMR frequency assignments are usually considered a prerequisite for the analysis of NOESY spectra, in turn required for the calculation of biomolecular structures. In contrast, as we propose here, relatively high numbers of unambiguous NOE identities can be consistently achieved in an automated manner by relying only on grouping resonances into connected spin systems. To achieve this goal, we have developed for proteins two protocols, SPI and BACUS, based on Bayesian inference. SPI (Grishaev and Llinas, 2002c) produces a list of the (1)H resonance frequencies from homo- and hetero-nuclear multidimensional spectra, grouped into effective spin systems. BACUS automatically establishes probabilistic identities of NOESY cross-peaks in terms of the chemical shifts provided by SPI. BACUS requires neither assignment of resonances nor an initial structural model. It successfully copes with chemical shift overlap and does so without cycling through 3D structure calculations. The method exploits the self consistency of the NOESY graph by taking advantage of a network of J- as well as NOE-connected "reporter" protons sorted via SPI. BACUS was validated by tests on experimental NOESY data recorded for the col 2 and kringle 2 domains. PMID- 14739636 TI - Rotational-echo double-resonance NMR-restrained model of the ternary complex of 5 enolpyruvylshikimate-3-phosphate synthase. AB - The 46-kD enzyme 5-enolpyruvylshikimate-3-phosphate (EPSP) synthase catalyzes the condensation of shikimate-3-phosphate (S3P) and phosphoenolpyruvate to form EPSP. The reaction is inhibited by N-(phosphonomethyl)-glycine (Glp), which, in the presence of S3P, binds to EPSP synthase to form a stable ternary complex. We have used solid-state NMR and molecular modeling to characterize the EPSP synthase-S3P Glp ternary complex. Modeling began with the crystal coordinates of the unliganded protein, published distance restraints, and information from the chemical modification and mutagenesis literature on EPSP synthase. New inter ligand and ligand-protein distances were obtained. These measurements utilized the native (31)P in S3P and Glp, biosynthetically (13)C-labeled S3P, specifically (13)C and (15)N labeled Glp, and a variety of protein-(15)N labels. Several models were investigated and tested for accuracy using the results of both new and previously published rotational-echo double resonance (REDOR) NMR experiments. The REDOR model is compared with the recently published X-ray crystal structure of the ternary complex, PDB code 1G6S. There is general agreement between the REDOR model and the crystal structure with respect to the global folding of the two domains of EPSP synthase and the relative positioning of S3P and Glp in the binding pocket. However, some of the REDOR data are in disagreement with predictions based on the coordinates of 1G6S, particularly those of the five arginines lining the binding site. We attribute these discrepancies to substantive differences in sample preparation for REDOR and X ray crystallography. We applied the REDOR restraints to the 1G6S coordinates and created a REDOR-refined xray structure that agrees with the NMR results. PMID- 14739637 TI - Improvement of hydrogen bond geometry in protein NMR structures by residual dipolar couplings--an assessment of the interrelation of NMR restraints. AB - We have examined how the hydrogen bond geometry in three different proteins is affected when structural restraints based on measurements of residual dipolar couplings are included in the structure calculations. The study shows, that including restraints based solely on (1)H(N)-(15)N residual dipolar couplings has pronounced impact on the backbone rmsd and Ramachandran plot but does not improve the hydrogen bond geometry. In the case of chymotrypsin inhibitor 2 the addition of (13)CO-(13)C(alpha) and (15)N-(13)CO one bond dipolar couplings as restraints in the structure calculations improved the hydrogen bond geometry to a quality comparable to that obtained in the 1.8 A resolution X-ray structure of this protein. A systematic restraint study was performed, in which four types of restraints, residual dipolar couplings, hydrogen bonds, TALOS angles and NOEs, were allowed in two states. This study revealed the importance of using several types of residual dipolar couplings to get good hydrogen bond geometry. The study also showed that using a small set of NOEs derived only from the amide protons, together with a full set of residual dipolar couplings resulted in structures of very high quality. When reducing the NOE set, it is mainly the side-chain to side chain NOEs that are removed. Despite of this the effect on the side-chain packing is very small when a reduced NOE set is used, which implies that the over all fold of a protein structure is mainly determined by correct folding of the backbone. PMID- 14739638 TI - An evaluation of detergents for NMR structural studies of membrane proteins. AB - Structural information on membrane proteins lags far behind that on soluble proteins, in large part due to difficulties producing homogeneous, stable, structurally relevant samples in a membrane-like environment. In this study 25 membrane mimetics were screened using 2D (1)H-(15)N heteronuclear single quantum correlation NMR experiments to establish sample homogeneity and predict fitness for structure determination. A single detergent, 1-palmitoyl-2-hydroxy-sn-glycero 3-[phospho-RAC-(1-glycerol)] (LPPG), yielded high quality NMR spectra with sample lifetimes greater than one month for the five proteins tested - R. sphaeroides LH1 alpha and beta subunits, E. coli and B. pseudofirmus OF4 ATP synthase c subunits, and S. aureus small multidrug resistance transporter - with 1, 2, or 4 membrane spanning alpha-helices, respectively. Site-specific spin labeling established interhelical distances in the drug transporter and genetically fused dimers of c subunits in LPPG consistent with in vivo distances. Optical spectroscopy showed that LH1 beta subunits form native-like complexes with bacteriochlorophyll a in LPPG. All the protein/micelle complexes were estimated to exceed 100 kDaltons by translational diffusion measurements. However, analysis of (15)N transverse, longitudinal and (15)N[(1)H] nuclear Overhauser effect relaxation measurements yielded overall rotational correlation times of 8 to 12 nsec, similar to a 15-20 kDalton protein tumbling isotropically in solution, and consistent with the high quality NMR data observed. PMID- 14739639 TI - New applications of 2D filtered/edited NOESY for assignment and structure elucidation of RNA and RNA-protein complexes. AB - NMR spectra of large RNAs are difficult to assign because of extensive spectral overlap and unfavorable relaxation properties. Here we present a new approach to facilitate assignment of RNA spectra using a suite of four 2D-filtered/edited NOESY experiments in combination with base-type-specific isotopically labeled RNA. The filtering method was developed for use in 3D filtered NOESY experiments (Zwahlen et al., 1997), but the 2D versions are both more sensitive and easier to interpret for larger RNAs than their 3D counterparts. These experiments are also useful for identifying intermolecular NOEs in RNA-protein complexes. Applications to NOE assignment of larger RNAs and an RNA-protein complex are presented. PMID- 14739640 TI - New NMR experiments for RNA nucleobase resonance assignment and chemical shift analysis of an RNA UUCG tetraloop. PMID- 14739641 TI - Solution structure of hypothetical protein TA1414 from Thermoplasma acidophilum. PMID- 14739642 TI - Backbone 1H, 13C and 15N resonance assignments of the response regulator HP1043 from Helicobacter pylori. PMID- 14739643 TI - 1H, 13C and 15N resonance assignments for the N-cadherin prodomain. PMID- 14739644 TI - Assignments of 1H and 15N resonances of the bacteriophage lambda capsid stabilizing protein gpD. PMID- 14739645 TI - Backbone 1H, 15N and 13C assignments for the 21 kDa Caenorhabditis elegans homologue of "brain-specific" protein. PMID- 14739646 TI - Backbone resonance assignment of an aminoglycoside-3'-phosphotransferase type IIa. PMID- 14739647 TI - 1H, 13C and 15N backbone resonance assignments of the dimeric yeast peroxiredoxin YLR109w. PMID- 14739648 TI - The changes in the T helper 1 (Th1) and T helper 2 (Th2) cytokine balance during HIV-1 infection are indicative of an allergic response to viral proteins that may be reversed by Th2 cytokine inhibitors and immune response modifiers--a review and hypothesis. AB - The HIV-1 infection in humans induces an early cellular immune response to react to the viral proteins with a cytotoxic T cell (CTL) response that fails to inhibit virus replication and the spread of the virus. It became evident that the progression of the disease causes chronic changes to the immune system of which a gradual increase in IgE antibodies is one of its features. When the HIV-1 epidemic began, the relation between the gradual increase in IgE content and AIDS was not understood, but later it became a marker for disease prognosis. The advances in the knowledge on T helper 1 (Th1) and T helper 2 (Th2) cells revealed that Th1 cells produce cytokines that stimulate the proliferation of CTLs. Th2 cells produce cytokines that are responsible for the activation of the humoral immune response in healthy people. Studies on both Th1 and Th2 cytokine synthesis revealed an aberration in HIV-1 infected people. Clerici and Shearer presented a hypothesis (1993) whereby Th1 cell activity declines and Th2 activity increases (the Th1 --> Th2 switch hypothesis) in HIV-1 infected people. In fact, experiments concerning this hypothesis ultimately supported the premise that the switch involves a critical change in the cytokine balance, which leads to the contraction of AIDS. However, the research community must still discern why such a Th1 --> Th2 switch takes place in infected people and how it can be reversed. The present review points to the fact that a similar Th1 --> Th2 switch constitutes the response of allergic people to environmental allergens. HIV-1 patients and allergic people that are exposed to allergens respond with an increased synthesis of Th2 cytokines and IgE, together with a decrease in Th1 cytokines. The studies on allergen-induced Th2 cells revealed that the Th2 cytokine IL-4 induces B cells to synthesize IgE, and cytokine IL-5 is the inducer of eosinophilia, just as in HIV-1 infection. The difference between the HIV-1 infection and allergies is the ability of IL-4 to induce the synthesis in T cells of the HIV-1 coreceptor CXCR4 that selects from the replicating virus a syncytium inducing (SI) virus, a variant virus that replicates rapidly. The present hypothesis implicates the viral proteins in the induction of Th2 cytokine synthesis. This suggests that in viral proteins, allergen-like domains may be responsible for the activation of Th2 cytokine synthesis. Based on the analogy of the responses of humans to allergens and HIV-1, the following hypotheses is suggested: (a) Removal of allergen-like domains from viral genes by genetic engineering may provide viral proteins for vaccine development. (b) Attempts to treat allergic patients with IL-4 receptor inhibitors suggests that the "Th2 --> Th1 Reversion" constitutes a possible approach to inhibiting the Th2 cytokines and inducing a revival of the anti-viral Th1 response. PMID- 14739649 TI - Investigations on the ORF 167L of lymphocystis disease virus (Iridoviridae). AB - The predicted open reading frame 167L (ORF 167L) of lymphocystis disease virus (LCDV, Iridoviridae ) isolated from plaice, dab and flounder was investigated. The ORF 167L corresponding genes of the three LCDV isolates were amplified, cloned and sequenced. A comparison of the LCDV strains showed that the nucleotide sequence of ORF 167L and its deduced amino acid sequence were highly conserved in the genus lymphocystivirus (a homology of 80% in dab and flounder/plaice, 97% in plaice and flounder). The N-terminus protein predicted from the ORF 167L suggests similarities to the tumor necrosis factor receptor (TNFR)-family, and to TNFR like proteins, which play an important role in various poxvirus species. Further, homology to the CUB-domain was shown at the C-terminus of the LCDV protein. Phylogenetic analyses of partial LCDV protein sequences identified two clusters: one cluster containing the flounder and plaice LCDV isolate (LCDV-1), and another cluster, containing the dab LCDV isolate (LCDV-2). The ORF 167L of plaice LCDV was expressed in Escherichia coli, and in fish cells. The expressed ORF resulted in a 30-kDa cytoplasmic protein lacking a signal peptide. An established monoclonal antibody (mAb 18) was used to detect LCDV proteins in skin explants of flounders and cryosections of dab skin. Specific fluorescence was found in the cytoplasm of intact epitheloid cells of the lymphocystis capsule and in the epidermis skin covering the lymphocystic nodules. LCDV-specific labelling of mAb 18 was also shown in spleen and liver tissue of LCDV-positive flounders. The ORF 167L protein seemed not to have the extracellular receptor function predicted from the usual cellular TNFR. The myxomavirus M-T2 protein, a poxviral TNFR homologue, was also shown not to have TNFR-like functions but to be involved in the apoptosis signal cascade. PMID- 14739650 TI - Base usage and dinucleotide frequency of infectious bursal disease virus. AB - Base usage and dinucleotide frequency have been extensively studied in many eukaryotic organisms and bacteria, but not for viruses. In this paper, a comprehensive analysis of these aspects for infectious bursal disease virus (IBDV) was presented. The analysis of base usage indicated that all of the IBDV genes possess equivalent overall nucleotide distributions. However when the base usage at each codon positions was analysed by using cluster analysis, the VP5 open reading frame (ORF) formed a different cluster isolated from the other genes. The unusual base usage of VP5 ORF may indicate that the gene was originated by the virus "overprinting strategy", a strategy in which virus may create novel gene by utilizing the unused reading frames of its existing genes. Meanwhile, the GC content of the IBDV genes and the chicken's coding sequences was comparable; suggesting the virus imitation of the host to increase its translational efficiency. The analysis of dinucleotide frequency indicated that IBDV genome had dinucleotide bias: the frequencies of CpG and TpA were lower and the TpG was higher than the expected. Classical methylation pathway, a process where CpG converted to TpG, may explain the significant correlation between the CpG deficiency and TpG abundance. "Principal component analysis of the dinucleotide frequencies" (DF-PCA) was used to analyse the overall dinucleotide frequencies of IBDV genome. DF-PCA on the hypervariable region and polyprotein (VPX-VP4-VP3) gene showed that the very virulent IBDV (vvIBDV) was segregated from other strains; which meant vvIBDV had a unique dinucleotide pattern. In summary, the study of base usage and dinucleotide frequency had unravelled many overlooked genomic properties of the virus. PMID- 14739651 TI - Genetic rearrangements in the gC gene of the feline herpesvirus type 1. AB - In the field isolate, 91-58, of feline herpesvirus type 1 (FHV-1), one of the major immunogenic proteins was found to have different molecular masses of 75 and 130 kDa from those in the other field isolates (Maeda et al., J Vet Med Sci 57, 147-150, 1995). Immunoblot analysis using monoclonal antibodies (MAbs) indicated that the protein is glycoprotein C (gC). The gC gene of 91-58 was amplified by polymerase chain reaction (PCR) and shown to have an inserted fragment of approximately 160 base pairs (bp). Restriction endonuclease analysis of the PCR product with various restriction enzymes was carried out, indicating that the insertion located within 262 bp between Eco RV and DraI sites. Nucleotide sequence analysis indicated that the inserted fragment was 156 bp encoding 52 amino acids and composed repeat sequences. Next, five recent isolates were also examined by immunoblot analysis using anti-FHV-1 cat serum or MAbs. The result showed that one isolate, 98-064, also had the gC with different molecular weights. PCR and nucleotide sequence analyses indicated that 98-064 had an inserted sequence of 78 bp at the corresponding region identified in the gC gene of 91-58, although the inserted sequence was different from that of 91-58. These results indicated that some of FHV-1 isolates had the genetic rearrangements in the gC gene and detection of such mutations would be useful for differentiation among FHV-1 field isolates. PMID- 14739652 TI - Evidence of chromosomal integration of AAV DNA in human testis tissue. AB - Persistent infection with adeno-associated virus (AAV) has been demonstrated in human tissues, most frequently in the female and male genital tract. The clinical significance of latent AAV infection remains, however, uncertain to date. The mode of latency of AAV is not known, i.e., it is unclear whether the viral genome is integrated in the cellular genome, and if integration occurs site-specifically in chromosome 19 as has been observed in cell culture. Therefore we investigated if viral DNA in AAV DNA-positive human testis samples from two patients, is integrated in the cellular genome. Using two different molecular approaches, uni directional PCR and Walking Primer PCR, we could demonstrate that AAV DNA is present in an integrated form in testis tissue. Virus-cell DNA junction fragments were cloned and sequenced. A detailed analysis revealed integration within sequences of the so-called AAVS1 region on chromosome 19. These data demonstrate that AAV DNA can integrate also after natural infection, and that integration occurs within the AAVS1 region, at least in some cases. PMID- 14739653 TI - Simian cytomegalovirus encodes five rapidly evolving chemokine receptor homologues. AB - Many herpesviruses, poxviruses and retroviruses encode proteins related to chemokines and chemokine receptors. The first one discovered, US28 of human cytomegalovirus (HCMV), is a 7-transmembrane domain G protein-coupled chemokine receptor able to activate diverse cellular responses, including cell migration and gene expression. A related ORF named US27 is adjacent to US28, but no functions have been defined yet. Recently ORFs 3-7, a cluster of five concatenated ORFs with highest homology to US28 and mammalian chemokine receptors, were sequenced from a prototype "stealth virus", an African green monkey simian CMV (SCMV)-related entity with unusual fungal, bacterial and mammalian gene homologues. Stealth viruses have not yet been independently replicated in tissue culture, and therefore their biological significance remains unclear. ORF3, ORF4, ORF5 and ORF6 are complete ORFs whereas the sequence of ORF7 is incomplete. In the present study, we identified five corresponding ORFs in the genome of a clinical isolate of bonafide simian CMV (SCMV), strain 9610. We found substantial differences between the SCMV and "stealth virus" ORFs, especially for ORF5 where there are 31% non-identities at the amino acid level. Four conserved genes unrelated to chemokines (64K/CAP, DNBI, UL32, and IE2) in SCMV and HCMV had on average 52% identity at the deduced amino acid level, whereas the corresponding values for the SCMV ORFs versus US28 ranged from 21% to 30%, suggesting rapid gene diversification in this cluster. Consistent with this, the amino acid identity for any pairwise comparison among the SCMV ORFs is only 21 52%. The chemokine receptor homologues are estimated to comprise approximately 2 3% of the SCMV genome. HCMV US27 and US28 homologues have also been identified in the chimpanzee CMV genome, whereas mouse and rat CMV lack chemokine receptor homologues. This genomic analysis indicates that SCMV has an unusually high concentration of US28-related chemokine receptor homologues that have arisen by gene duplication and have diverged extensively from their closest relatives in mammals and other beta herpesviruses. The rate of divergence appears to be very rapid compared to other known SCMV genes, suggesting strong positive selection. PMID- 14739654 TI - Genetic vaccination of mice with plasmids encoding the NS1 non-structural protein from tick-borne encephalitis virus and dengue 2 virus. AB - Although there is a safe, inexpensive and efficacious vaccine against yellow fever, vaccination against other flavivirus diseases is less successful. There is no licensed vaccine against dengue fever and current vaccines against tick-borne encephalitis (TBE) and Japanese encephalitis are expensive and require several injections. Furthermore novel vaccines containing only virus envelope proteins may raise fears over antibody mediated enhancement (ADE) of disease. Here we report the successful use of genetic vaccination against TBE in an experimental animal model using a plasmid containing the coding sequence of a non-structural protein (NS1). Such vaccines would provide inexpensive protection against disease, without raising concerns over inducing ADE on subsequent exposure to heterotypic infectious virus. Attempts to generate chaemeric plasmids to protect against both TBE and dengue fever were less successful. Although these chaemeric plasmids directed the synthesis and secretion of the virus NS1 protein normally, no protection was observed against either TBE or dengue fever. PMID- 14739655 TI - Re-evaluation and in silico annotation of the Tupaia herpesvirus proteins. AB - Herpesviruses represent an exceptionally suitable model to analyze evolutionary old pathogens, their competency to adapt to existing and changing molecular niches in host species, and the modulation of the gene content and function to comply with the requirements of life. The basis for numerous studies dealing with these questions are reliable statements about the gene content of herpesviral genomes and the functions of viral proteins. The recent determination of the coding strategy of the chimpanzee cytomegalovirus genome and the re-evaluation of the gene content of the human cytomegalovirus genome made it also necessary to restructure the putative transcription map of the Tupaia herpesvirus (THV) genome. Twenty-three THV-specific ORFs formerly predicted to be coding for viral proteins were deleted from the THV transcription map resulting in a gene layout that is now characterized by the presence of conserved genes in the genome center, that probably reflect the genome structure of common herpesviral ancestors, and species-specific genes at the termini. The conserved regions in the THV genome are characterized by high G + C contents between 60% and 80%, a high CpG dinucleotide frequency, and the presence of densely packed putative CpG islands. The genome termini seem to provide the requirements of large scale rearrangements and complements of the gene content to adapt to new environmental demands. With the help of the recently designed method of dictionary-driven, pattern-based protein annotation it was possible to assign putative functions to almost all potential THV proteins, e.g. 123 were found to be putative membrane or secreted proteins, putative signal domains were identified in 69, and 29 proteins were predicted to be glycosylated. The present study adds new aspects to the knowledge about the precise gene composition of herpesvirus genomes and viral protein functions that are of exceptional importance for studies dealing with the phylogeny, the evolution, vaccine vector development, virus-host interactions, pathogenesis and the determination of protein functions of herpesviruses. PMID- 14739656 TI - Upregulation of LMP1 expression by histone deacetylase inhibitors in an EBV carrying NPC cell line. AB - OBJECTIVES: In about 60% of Epstein-Barr virus (EBV) carrying nasopharyngeal carcinomas (NPC) LMP1 expressing cells can be detected. The frequency of LMP1 positive cells and the expression level varies from cell to cell in the different tumors. Cell lines derived from EBV positive NPCs loose the virus during in vitro culture. The in vitro infected NPC cell line TWO3-EBV used in our study carries the neomycin-resistance gene containing EBV and expresses low level of LMP1. With this cell line it was thus possible to study the regulation of LMP1 expression by modification of chromatin acetylation state. STUDY DESIGN: The TWO-EBV cell line was treated with n -butyrate (NB) or trichostatin A (TSA). RESULTS: Shown by immunoblotting, the LMP1 level was elevated in the treated samples. Already 2 h after TSA exposure LMP1 expression was higher and it increased up to 24 h. Immunofluorescence staining showed that nearly all cells were LMP1 positive. Neither EBNA2 nor BZLF1 were induced. Tested first 2 h after the treatment, acetylated histone H3 and H4 were already detectable, and their level increased up to 8 h. Chromatin immunoprecipitation (ChIP) verified that the LMP1-promoter (LMP1p) (ED-L1) was acetylated after TSA treatment. CONCLUSION: EBV carrying epithelial cells do not express EBNA-2. We showed that LMP1 expression was upregulated by histone deacetylase inhibitors in an in vitro infected, EBV carrier NPC cell line. PMID- 14739657 TI - Intratypic variability of a tandem repeat locus within the DNA polymerase gene of human herpes simplex virus type 2. AB - In this study, the irntratypic variability of a tandem repeat locus within the DNA polymerase (pol) gene of human herpes simplex virus type 2 (HSV2) was uncovered. The locus contained variable numbers of tandem dodecanucleotide (5' GAC GAG GAC GGG-3') repetitive units. Our result showed that approximately 95% of analyzed HSV2 clinical isolates and the current GenBank HSV2 strains contained two copies of the repetitive units. From genital herpes specimens, three new HSV2 strains, which respectively contained 1, 3, and 4 copies of the repetitive units, were identified. This variable number of tandem repeat (VNTR) locus is absent in HSV1, and thus it also contributes to the intertypic variability of HSV1 and HSV2. The intratypic variability of the locus may be useful for HSV2 strain genotyping and this application is discussed. PMID- 14739658 TI - Complexity of dsRNA mycovirus isolated from Fusarium graminearum. AB - Fusarium graminearum is the causal agent of a serious scab disease of small grains in Korea. We screened 827 isolates of F. graminearum from diseased barley and maize and tested for the presence of double-stranded RNA (dsRNA) mycovirus. Of them, 19 isolates contained various sizes of dsRNAs. A dsRNA associated with pronounced morphological changes including reduction in mycelial growth, increase in dark orange to red pigmentation, reduced sporulation and virulence was previously observed in nine dsRNA-containing Fusarium isolates (Chu et al., Appl Env Microbiol 68, 2529-2534, 2002). Ten additional isolates were found infected with dsRNA mycoviruses. These mycoviruses contain 2-4 different segments of dsRNAs with the size-range of approximately 1.7-10 kbp in length. The presence of dsRNAs did not affect colony morphology and were transmissible through conidia and ascospore with incidence of 30-100%. Interestingly, dsRNA mycovirus found in F. graminearum isolates, JB33 and JNKY19, that show the pattern of mixed infection of two different viruses were transmitted to all progeny conidia and ascospores. These results indicate that there is genomic diversity of dsRNA mycoviruses that infect F. graminearum isolates and that impact of virus infection on host's morphology and virulence is determined by the interaction between dsRNAs and the fungal host, not by the mere presence of the dsRNAs. PMID- 14739659 TI - Resveratrol antagonizes EGFR-dependent Erk1/2 activation in human androgen independent prostate cancer cells with associated isozyme-selective PKC alpha inhibition. AB - The development of androgen-independent prostate cancer (AI PrCa) involves constitutive Erk1/2 activation sustained by the epidermal growth factor/transforming growth factor-alpha/EGF receptor (EGF/TGFalpha/EGFR) axis and other trophic signaling mechanisms in neoplastic human prostate epithelial cells in vivo. In this report, we show that growth-inhibitory concentrations of the dietary phytochemical resveratrol suppress EGFR-dependent Erk1/2 activation pathways stimulated by EGF and phorbol ester (12- O -tetradecanoyl phorbol 13 acetate, TPA) in human AI PrCa PC-3 cells in vitro. Because protein kinase C (PKC) is the major cellular receptor for phorbol esters and taking into consideration that resveratrol is PKC-inhibitory, we investigated resveratrol effects on cellular PKC isozymes associated with the suppression of TPA-induced Erk1/2 activation. The PKC isozyme composition of PC-3 cells was defined by Western analysis of the cell lysate with a comprehensive set of isozyme-selective PKC Ab's. PC-3 cells expressed PKCalpha, epsilon, zeta, iota, and PKD (PKCmicro), as did another human AI PrCa cell line of distinct genetic origin, DU145. The effects of resveratrol on TPA-induced PKC isozyme activation were defined by monitoring PKC isozyme translocation and autophosphorylation. Under conditions where resveratrol suppressed TPA-induced Erk1/2 activation, the phytochemical produced isozyme-selective interference with TPA-induced translocation of cytosolic PKCalpha to the membrane/cytoskeleton and selectively diminished the amount of autophosphorylated PKCalpha in the membrane/cytoskeleton of the TPA treated cells. These results demonstrate that resveratrol abrogation of a PKC mediated Erk1/2 activation response in PC-3 cells correlates with isozyme selective PKCalpha inhibition. The results provide evidence that resveratrol may have value as an adjuvant cancer therapeutic in advanced prostate cancer. PMID- 14739660 TI - The new isothiocyanate 4-(methylthio)butylisothiocyanate selectively affects cell cycle progression and apoptosis induction of human leukemia cells. AB - We investigated proliferation and apoptosis induction in Jurkat T-leukemia cells by the new isothiocyanate 4-(methylthio)butylisothiocyanate (MTBITC). To help elucidate whether the effects of MTBITC are specific for cancer cells, we tested MTBITC on freshly isolated, non-transformed human peripheral T lymphocytes. The effects of MTBITC are leukemic-cell-specific and consist of derangements in a critical point of cell-cycle control (G2/M transition). In fact, an increase in the proportion of G2 cells (from about 18% to 50%) was apparent following 24 h of treatment, associated with a decrease in the protein expression of cyclin B1. The expression of cyclin-dependent kinase (CDK) 1 was more mildly attenuated by MTBITC. Our results demonstrated that high concentrations of MTBITC can also induce apoptosis, through an increase of p53 and bax, but not bcl-2, protein expression. No effects of MTBITC were demonstrated on non-transformed T lymphocytes. Taking into account its in vitro antineoplastic activity and selectivity toward leukemia cells, MTBITC can be viewed as a conceptually promising agent in cancer therapy. PMID- 14739661 TI - Phase I trial of the polyamine analog N1,N14-diethylhomospermine (DEHSPM) in patients with advanced solid tumors. AB - BACKGROUND: This phase I study was conducted to determine maximal tolerated dose (MTD) and dose-limiting toxicities (DLT) in patients with advanced solid tumors treated with the polyamine analog N1, N14-diethylhomospermine (DEHSPM). METHODS: Patients were treated with DEHSPM administered as a subcutaneous (SC) injection daily for five consecutive days repeated every 4 weeks. Three dose levels were examined starting at 12.5 mg/m2/day, escalating to 37.5 mg/m2/day. RESULTS: A total of 15 patients were enrolled. Dose limiting toxicities (grade 3 or 4) included nausea, vomiting, constipation, ileus, elevations of aspartate aminotransferase (AST) and alkaline phosphatase, hyperbilirubinemia, and ventricular bigeminy. CONCLUSION: DEHSPM given as a SC injection is overall well tolerated at lower doses, but significant toxicities were observed at the 37.5mg/m2/day dose level. MTD was established at 25 mg/m2/day but further investigation with this study drug is not recommended secondary to the potential for neurotoxicities and hepatic damage as a result of cumulative doses. PMID- 14739662 TI - A phase I trial of daily oral 4'- N -benzoyl-staurosporine in combination with protracted continuous infusion 5-fluorouracil in patients with advanced solid malignancies. AB - PURPOSE: 4'- N -Benzoyl-staurosporine (PKC412) is an orally available staurosporine derivative that inhibits protein kinase C. The objectives of this phase I trial were to determine the maximum tolerated dose (MTD), the dose limiting toxicities (DLTs), and the pharmacokinetics of PKC412 when co administered with 5-Fluorouracil (5-FU). EXPERIMENTAL DESIGN: PKC412 was given daily with a 21-day continuous i.v. infusion of 5-FU 200 mg/m2/day, repeated every 4 weeks. The PKC412 dose was escalated by a modified continual reassessment method. The steady-state plasma pharmacokinetics of 5-FU, PKC412, and two of its circulating metabolites were determined during the first cycle of therapy. RESULTS: A total of 33 patients were treated with 70 cycles (median: 2, range: 1 4) of PKC412 at doses ranging from 25 to 225 mg/day. No significant toxicities were encountered with doses up to 150 mg/day. Among nine patients treated with 225 mg/day of PKC412, one experienced grade 3 fatigue and nausea, another developed grade 3 hyperglycemia, and three had grade 2 emesis and stomatitis, leading to early treatment discontinuation. Minor responses consisting of a 40 45% tumor reduction were observed in two patients, one with gall bladder carcinoma and one with breast cancer. Mean values of steady-state pharmacokinetic variables for both PKC412 and 5-FU were comparable to single agent studies. CONCLUSIONS: The recommended phase II dose of PKC412 is 150 mg/day when combined with a continuous infusion of 200 mg/m2/day 5-FU. The dose limiting toxicity was grade 2 emesis and stomatitis and the regimen showed indications of activity. There was no evidence of a pharmacokinetic interaction between the two drugs. PMID- 14739663 TI - Pharmacokinetic drug-drug interaction of the novel anticancer agent E7070 and acenocoumarol. AB - E7070 is a novel sulfonamide anticancer agent that arrests cancer cells at the G1/S boundary of the cell cycle. Three patients receiving chronic therapy with the oral anticoagulant acenocoumarol experienced bleeding and/or a prolonged prothrombin time after treatment with E7070 at a dose of 700 mg/m2 given as a 1-h infusion. In vitro studies have shown that E7070 has the potential to inhibit several cytochrome P450 (CYP)-enzymes, including CYP2C9, CYP2C19, CYP2D6, CYP2E1, and CYP3A4. The major enzyme involved in the metabolism of acenocoumarol in man is CYP2C9. This study was performed to investigate the interaction between E7070 and acenocoumarol. Blood samples were obtained from two patients receiving daily oral maintenance treatment with acenocoumarol both prior to and following treatment with E7070. In addition, we incubated acenocoumarol enantiomers with pooled human microsomes with and without E7070 and measured the in vitro plasma protein binding of acenocoumarol after incubation with E7070. Pharmacokinetic parameters of acenocoumarol were calculated by noncompartmental analysis and revealed that in both patients the area under the concentration-time curve up to 24 h after the acenocoumarol administration was higher following E7070 (2.56 and 1.58 h*micromol/L) compared to the systemic exposure in the absence of E7070 (1.87 and 1.23 h*micromol/l). The formation of acenocoumarol metabolites was retarded by E7070 at already low concentrations (2.1 microM). The plasma protein binding of acenocoumarol was reduced at higher concentrations of E7070 (259 microM). These results indicate that E7070 may primarily interact with acenocoumarol by reducing its systemic clearance. Displacement of acenocoumarol's plasma protein binding by E7070 may also occur but to a minor extent. In the absence of careful monitoring this drug-drug interaction may result in hypoprothrombinemia and a hemorrhagic tendency. PMID- 14739664 TI - Phase I and pharmacokinetic study of fostriecin given as an intravenous bolus daily for five consecutive days. AB - Fostriecin (CI-920) is a potent inhibitor of protein phosphatase 2A (PP2A) and protein phosphatase 4(PP4) found to have anticancer activity in preclinical testing. A phase I study was conducted to evaluate the maximum-tolerated dose (MTD), toxicity profile, and pharmacokinetics (PK) of this drug. Forty-six patients were treated with escalating doses of fostriecin (2-47 mg/m2) administered as a daily bolus infusion for five consecutive days. PK studies were performed at different time points following administration of fostriecin. Dose limiting toxicities included: elevation of creatinine, bilirubin, and hepatic transaminases; nausea, anorexia, lethargy, and hypotension. PK studies were compatible with a two-compartment model. Regression analysis revealed a significant relationship between dose and clearance; however, the r2 value was only 0.168 indicating a low predictive value for the model. No significant difference was seen in PK parameters with repeated dosing during the same cycle. Although no tumor responses were seen, 16 patients had stable disease with a median duration response of 2.6 months. The study was closed before reaching MTD due to problems with the supply of fostriecin from the National Cancer Institute of the United States (NCI US). New methods for synthesizing fostriecin have recently been described and therefore further development of this unique anticancer agent may be warranted. PMID- 14739665 TI - Phase II trial of intra-arterial chemotherapy using a novel lipophilic platinum derivative (SM-11355) in patients with hepatocellular carcinoma. AB - SM-11355, a lipophilic platinum derivative, is a novel intra-arterial chemotherapeutic agent for hepatocellular carcinoma (HCC). A phase II study of SM 11355 was conducted to evaluate the antitumor activities and the toxicity in chemotherapy-naive patients with HCC. Sixteen patients were treated with transcatheter arterial injection of SM-11355-lipiodol emulsion (20-120 mg/body). The responses were evaluated by computed tomography 3 months after treatment. Complete response (CR) was defined as disappearance or 100% necrosis of all tumors, and lipiodol accumulation in tumors was regarded as indicating necrosis. Nine patients achieved CR (56%; 95% confidence interval, 29.9-80.2%). The grade 3 toxicities were neutropenia (19%), total bilirubin elevation (19%), AST elevation (44%), and ALT elevation (19%). None of the patients showed grade 4 toxicities or episodes of renal dysfunction. Other common adverse effects were eosinophilia (100%) and pyrexia (94%). Intra-arterial chemotherapy with SM-11355, which was well tolerated, showed promising antitumor activity in patients with HCC. PMID- 14739666 TI - Phase II trial of temozolomide in patients with cisplatin-refractory germ cell tumors. AB - Fourteen patients with cisplatin-refractory germ cell tumors (GCT) were treated with temozolomide on a phase II trial. Temozolomide was given orally at 150 mg/m2/day on days 1-5. The cycle length was 28 days. No patient experienced a grade 3 or 4 toxicity, and none of the 14 evaluable patients achieved a complete or partial response. Temozolomide is not efficacious in the treatment of cisplatin-refractory GCT patients. PMID- 14739667 TI - Phase II study of CI-958 in patients with hormone refractory prostate carcinoma. AB - A phase II trial of CI-958 (NSC #635371), a new benzothiopyranoindazole was performed in patients with hormone refractory prostate carcinoma using prostate specific antigen (PSA) levels for response assessment. Twenty-two patients were entered on this study and twenty one were eligible. Toxicity consisted mainly of granulocytopenia (71% grade 3 or 4), but there were no significant infections. Two patients were removed from study due to asymptomatic decreases in cardiac ejection fraction. Of 21 evaluable patients, there were four responders (19%, CI 0-35%). PMID- 14739668 TI - A novel combination of cisplatin, irinotecan, and capecitabine in patients with advanced cancer. AB - BACKGROUND: We conducted a dose escalation study combining cisplatin, irinotecan, and capecitabine (CIC), aiming to establish the maximum tolerated doses (MTD), side effect profile, and dose-limiting toxicity (DLT) of this novel regimen. PATIENTS AND METHODS: Intravenous cisplatin and irinotecan were to be administered on days 1 and 8, and oral capecitabine on days 1-14 of a 3-week cycle. The study was conducted in three parts. Part A: escalating doses of irinotecan (40 --> 80 mg/m2) and capecitabine (1000 --> 3300 mg/d) combined with a fixed dose of cisplatin (30 mg/m2). Part B: escalating doses of irinotecan (MTD A --> MTD-A + 40 mg/m2) with fixed doses of cisplatin (20 mg/m2) and capecitabine (MTD-A level). Part C: escalating doses of capecitabine (1300 mg/d-->2600 mg/d) with fixed doses of cisplatin (20 mg/m2) and irinotecan (60 mg/m2). RESULTS: Of 51 eligible patients 27 (53%) were male, median age was 58 years and 88% had PS 0 1. Major primary disease sites were colorectal (24%), unknown (14%), stomach (14%), and pancreas (12%). MTD-A was cisplatin 30 mg/m2, irinotecan 60 mg/m2, capecitabine 1000 mg/d and MTD-B was cisplatin 20 mg/m2, irinotecan 90 mg/m2, capecitabine 1000 mg/d. An MTD was not formally established for part C. DLTs consisted of infection with neutropenia (1), diarrhea and fatigue (1), hypokalemia (1), diarrhea and febrile neutropenia (1) and C2 delay of > or = 2 weeks or 25% dose reduction in C1 due to neutropenia or thrombocytopenia (6). Seven patients had a partial response to treatment (four colorectal, one SCLC, one NSCLC, one unknown primary), twenty seven SD (53%), twelve PD (24%) and five NE (10%). CONCLUSION: CIC was associated with moderate toxicity and only modest antitumor activity. We conclude that this regimen has insufficient activity to justify further study in the phase II setting. PMID- 14739669 TI - Weekly gemcitabine for the treatment of biliary tract and gallbladder cancer. AB - OBJECTIVES: To evaluate the efficacy and safety of weekly administration of gemcitabine treatment in chemotherapy-naive patients with advanced biliary tract and gallbladder cancer. PATIENTS AND METHODS: Gemcitabine at a dose of 800 mg/m2 was administered weekly as a 30-min infusion to patients with previously operated, histologically confirmed, metastatic, or unresectable locally advanced cholangiocarcinoma. Treatment was continued until unacceptable toxicity or disease progression. RESULTS: A total of 30 patients (median age 66 years; range 54-72 years) were included in the study. A median of 14 (range, 4-33) weekly doses was administered. Out of 30 patients evaluable for response, nine partial responses were observed (30.0%), while a further 11 patients demonstrated stable disease (36.7%). The median time to disease progression was 7 months (range, 5 34). Overall response rate was superior in patients with cancer of the gallbladder (ORR = 35.7%) compared with those patients with biliary duct cancer (ORR = 27.3%). This correlated to a significantly longer time to progression of 6.4 months (95% confidence interval (CI), 5.6-7.1 months) versus 3.6 months (95% CI, 2.9-4.3 months; p = 0.03) and a significantly better overall survival of 17.1 months (95% CI, 15.8-18.5 months) versus 11.4 months (95% CI, 10.2-12.6 months, p = 0.021). Toxicities were generally mild with only one case of grade 3 neutropenia. There were no cases of febrile neutropenia and no treatment-related deaths. CONCLUSIONS: Weekly administration of gemcitabine provides a safe, well tolerated, and effective treatment for chemotherapy naive patients with advanced cholangiocarcinoma, particularly with a gallbladder origin. PMID- 14739670 TI - Molecular methods clarify morphometric variation in triactinomyxon spores (Myxozoa) released from different oligochaete hosts. AB - Thirty-nine freshwater tubificid oligochaetes were isolated, each of which harboured a triactinomyxon infection. Spore characteristics include the typical triactinomyxon anchor shape, eight germ cells within the sporoplasm and three unequal (two long and one shorter) caudal processes with square tips. Despite morphological similarities between the spores from the different hosts, their morphometrical data varied considerably; significantly, the ranges of dimensions of the smallest and largest exemplars were mutually exclusive. In order to ascertain the true number of spore types present, molecular methods were employed. Samples of waterborne spores, including the smallest and largest representatives, were selected from 11 host oligochaetes (all Tubifex tubifex Muller) and a nested PCR-RFLP 'riboprint' analysis performed. The small subunit ribosomal DNA gene (18S rDNA) was targetted and amplified through two rounds of PCR, then digested with the restriction enzymes Dde I and Hha I. The resultant major cleavage patterns produced by both enzymes indicated a single triactinomyxon form; however, the pattern of several less intense bands varied between the samples. From a subset of five samples drawn from across the full spectrum of spore sizes, a 327 bp region near the 5' was sequenced and was identical for all five samples. Comparison of this 327 bp region with that of 12 other triactinomyxons in GenBank showed 68.7-96.9% similarity (at least 9 base differences). A further 469 bp generated for each of the smallest, largest and mid-range (= reference) spore samples was identical also. The reference sample was sequenced further to yield 1,554 bp of 18S rDNA (GenBank accession number AY162270); comparison with other Myxozoa indicated this sequence was novel. The morphometrics of our triactinomyxon did not correlate with any published description. The morphometrical variation exhibited by spores of the triactinomyxon type in this study raises questions about the validity of using morphometrical data to distinguish spore types and suggests that there could be taxonomic redundancy in the diversity of actinosporeans recorded in the literature. The additional information provided by molecular data in this study was pivotal in the clarification of morphometrical variation exhibited by morphologically similar triactinomyxon spores released from different oligochaete hosts. PMID- 14739671 TI - Some comments on the genus Lernaeopodina Wilson, 1915 (Copepoda: Siphonostomatoida: Lernaeopodidae). AB - The status of Lernaeopodina Wilson, 1915 is discussed. It is considered that L. spinacis (Brian, 1908) is a species inquirenda and that the transfer of L. pectinata Ho, 1985 to Kabatahoia Kazachenko, 2001 was justified. The genus is currently restricted to four species. PMID- 14739672 TI - Two species of parasitic copepods (Caligidae) new to Japan. AB - Two species of parasitic copepods (Siphonostomatoida: Caligidae), Caligus sclerotinosus Roubal, Armitage & Rohde, 1983, parasitic on red seabream Pagrus major (Temminck & Schlegel), and Lepeophtheirus longiventralis Yu & Wu, 1932, parasitic on spotted halibut Verasper variegatus (Temminck & Schlegel), are redescribed based on material found on their respective hosts cultured in Japan. Both species are new to Japan. Preliminary observation on the occurrence of L. longiventralis indicates that the larval development takes place on the body surface of the host and only the post-mated female migrates into the host's gill cavities. Whilst the occurrence of L. longiventralis in Japan can be considered as due to natural causes, the occurrence of C. sclerotinosus is likely due to anthropogenic activities. PMID- 14739673 TI - The identity of Limnoncaea diuncata Kokubo, 1914 (Copepoda: Poecilostomatoida) from Hokkaido, Japan, with the relegation of Diergasilus Do, 1981 to a junior synonym of Thersitina Norman, 1905. AB - Both sexes of an ergasilid copepod, Limnoncaea diuncata Kokubo, 1914, are redescribed based on planktonic specimens collected from the type-locality in Hokkaido, Japan. Comparison of this species with Thersitina gasterostei (Pagenstecher, 1861) revealed that they are conspecific. Another ergasilid genus with two claws on the antenna, Diergasilus Do, 1981, is relegated to synonymy with Thersitina Norman, 1905. The diagnosis of Thersitina is amended. PMID- 14739674 TI - New species of Tereancistrum Kritsky, Thatcher & Kayton, 1980 (Monogenea: Dactylogyridae: Ancyrocephalinae) from the gills of Prochilodus lineatus (Osteichthyes: Prochilodontidae) from the upper Parana River floodplain, Brazil. AB - Two new species of Tereancistrum Kritsky, Thatcher & Kayton, 1980 are described from the gills of Prochilodus lineatus (Prochilodontidae) collected from the upper Parana River floodplain, Brazil. T. toksonum n. sp. is characterised by a slim ventral bar with a sclerotised membrane along the anterior margin. Another characteristic is the dorsal anchor with an elongated deep root. T. curimba n. sp. is similar to T. ornatus Kritsky, Thatcher & Kayton, 1980 but possesses a sclerotised structure between the accessory sclerites of the ventral anchors. Moreover, the accessory anchor sclerites of this new species are longer than those described for T. ornatus. PMID- 14739675 TI - A new species of Nasicola Yamaguti, 1968 (Monogenea: Capsalidae) from the nasal cavities of Thunnnus obesus and a redescription of N. klawei (Stunkard, 1962) from T. albacares off Brazil. AB - Two species of Nasicola Yamaguti, 1968 are described from the nasal cavities of tunas ( Thunnus spp.) from off the coast of Brazil: N. brasiliensis n. sp. from T. obesus (Lowe) and the type-species, N. klawei (Stunkard, 1962), from T. albacares (Bonn.). The new species is differentiated from N. klawei on the basis of the large number of testes and from N. hogansi Wheeler & Beverley-Burton, 1986 by its greater body-size, proportionately smaller haptor and smaller number of marginal spines. The host-specificity of Nasicola spp. is commented upon. PMID- 14739676 TI - Moravecia australiensis n. g., n. sp. (Dracunculoidea: Guyanemidae) from the gills of the green porcupine fish Tragulichthys jaculiferus (Cuvier) in Australia. AB - A new dracunculoid genus and species, Moravecia australiensis, is described from gill-filaments of the green porcupine fish Tragulichthys jaculiferus (Cuvier) (Tetraodontiformes: Diodontidae) from Moreton Bay, Queensland, Australia. Abundant mobile larvae and a few adult males with females occurred in the gill filament between the epithelial basement membrane and efferent artery. Gills of all 69 fish examined contained larvae. Eleven harboured adult nematodes of a previously undescribed species belonging to the family Guyanemidae. The new species is placed within a newly proposed genus because it differs from the four existing genera in the family in possessing fine cuticular transverse striations, two forward protruding cephalic elevations, a circumoral elevation, a small triangular mouth surrounded by six cephalic papillae arranged in two lateral clusters of three each and a pair of large lateral amphids. Males have two pairs of pedunculate caudal papillae supporting the caudal alae. A key to the genera of the Guyanemidae is presented. PMID- 14739677 TI - The genus Biuterina Fuhrmann, 1902 (Cestoda, Paruterinidae) in the Old World: redescriptions of three species from Palaearctic Passeriformes. AB - The syntypes of Biuterina passerina Fuhrmann, 1908 from Alauda arvensis and Galerida cristata (Passeriformes, Alaudidae) from an unknown locality are redescribed. B. fuhrmanni Schmelz, 1941 is redescribed on the basis of its syntypes from Emberiza aureola from China; its type-material contains, in addition to a scolex and pre-gravid and gravid fragments of Biuterina, fragments of mature proglottides from a dilepidid cestode, which were erroneously used in the original description. Specimens, which correspond morphologically to B. clerci Spasskii, 1946, from Miliaria calandra, E. citrinella and E. cirlus from Bulgaria and from E. citrinella from the Czech Republic, are studied. The synonymy of B. clerci with B. passerina is rejected; however, it is recognised as a synonym of B. fuhrmanni. Available data suggest that B. passerina is a specific parasite of birds of the family Alaudidae, while B. fuhrmanni is specific to the Emberizidae. B. collurionis Matevosyan, 1950 is considered a species inquirenda, pending confirmation of apparent differences from B. passerina and B. fuhrmanni based on further material. Biuterina cordifera Murai & Sulgostowska, 1983 is redescribed on the basis of specimens, previously identified by Rysavy (1965) as B. triangula (Krabbe, 1869), from Acrocephalus scirpaceus (Muscicapidae, Sylviinae) in the Czech Republic (new geographical record) and from Erithacus megarhynchos (Muscicapidae, Turdinae) in Bulgaria (new host and geographical records). PMID- 14739678 TI - Newborn screening for cystic fibrosis: science, legislation, and human values. PMID- 14739679 TI - Neonatal screening for cystic fibrosis: France rises to the challenge. AB - This paper describes the adjustments to the French neonatal screening programme required by the introduction of systematic screening for cystic fibrosis (CF), taking into account both the legal and statutory framework and the lessons of a pilot study carried out 10 years ago. The French association for the screening and prevention of infant handicaps (AFDPHE) has been mandated by its regulatory agencies to organize screening for CF in France (metropolitan and overseas territories). During the year 2001, expert groups (Technical Aspects, Information, Ethics and Genetics, Criteria for CF Centres, Protocol for the Care of a Newborn with CF) issued recommendations for the establishment of a national programme that would guarantee efficiency and adequate patient care from the time of diagnosis onward. The programme is based on a strategy combining immunoreactive trypsin (IRT) assay and the analysis of DNA mutations in dried blood samples obtained at 3 days of age. When an elevated IRT value is found, DNA analysis is performed on the same sample. Owing to the relative regional heterogeneity existing in France, 30 selected mutations are used, which provide 85% coverage. The Ethics and Genetics Committee recommended that, in order to avoid arousing anxiety by a recall, informed consent, according to the French legislation on bioethics, should be obtained for all neonates at birth by having the parents sign directly on the sampling paper. Information brochures for parents and health professionals have been designed. A new organization of patient care, involving the creation of CF centres recognized by the Ministry of Health, has been decided; all children diagnosed are to be referred to such centres, where they can be well cared for by a trained staff with sufficient means. The programme was implemented region by region in France, from the beginning of the year 2002 to early 2003. The expert groups still meet periodically to evaluate the implementation of the programme and to check that the terms of the agreement between the AFDPHE and the Social Security Agency are complied with. PMID- 14739680 TI - Glycine N -methyltransferase deficiency: a new patient with a novel mutation. AB - We report studies of a Greek boy of gypsy origin that show that he has severe deficiency of glycine N -methyltransferase (GNMT) activity due to apparent homozygosity for a novel mutation in the gene encoding this enzyme that changes asparagine-140 to serine. At age 2 years he was found to have mildly elevated serum liver transaminases that have persisted to his present age of 5 years. At age 4 years, hypermethioninaemia was discovered. Plasma methionine concentrations have ranged from 508 to 1049 micro mol/L. Several known causes of hypermethioninaemia were ruled out by studies of plasma metabolites: tyrosinaemia type I by a normal plasma tyrosine and urine succinylacetone; cystathionine beta synthase deficiency by total homocysteine of 9.4-12.1 micro mol/L; methionine adenosyltransferase I/III deficiency by S -adenosylmethionine (AdoMet) levels elevated to 1643-2222 nmol/L; and S -adenosylhomocysteine (AdoHcy) hydrolase deficiency by normal AdoHcy levels. A normal plasma N -methylglycine concentration in spite of elevated AdoMet strongly suggested GNMT deficiency. Molecular genetic studies identified a missense mutation in the coding region of the boy's GNMT gene, which, upon expression, retained only barely detectable catalytic activity. The mild hepatitis-like manifestations in this boy are similar to those in the only two previously reported children with GNMT deficiency, strengthening the likelihood of a causative association. Although his deficiency of GNMT activity may well be more extreme, his metabolic abnormalities are not strikingly greater. Also discussed is the metabolic role of GNMT; several additional metabolite abnormalities found in these patients; and remaining questions about human GNMT deficiency, such as the long-term prognosis, whether other individuals with this defect are currently going undetected, and means to search for such persons. PMID- 14739681 TI - Homocystinuria due to cystathionine beta-synthase deficiency: novel biochemical findings and treatment efficacy. AB - To explore the pathogenesis of cystathionine beta-synthase (CBS) deficiency and to test the efficacy of pharmacological therapy we examined a panel of metabolites in nine homocystinuric patients under treated and/or untreated conditions. Off pharmacological treatment, the biochemical phenotype was characterized by accumulation of plasma total homocysteine (median 135 micromol/L) and blood S -adenosylhomocysteine (median 246 nmol/L), and by normal levels of guanidinoacetate and creatine. In addition, enhanced remethylation was demonstrated by low serine level (median 81 micromol/L), and by increased concentration of methionine (median 76 micromol/L) and N -methylglycine (median 6.8 micromol/L). Despite the substantially blocked transsulphuration, which was evidenced by undetectable cystathionine and severely decreased total cysteine levels (median 102 micromol/L), blood glutathione was surprisingly not depleted (median 1155 micromol/L). In 5 patients in whom pharmacological treatment was withdrawn, the differences of median plasma total homocysteine levels (125 micromol/L after withdrawal versus 33 micromol/L under treatment conditions), total cysteine levels (139 versus 211 micromol/L) and plasma serine levels (53 versus 103 micromol/L) on and off treatment demonstrated the efficacy of long term pyridoxine/betaine administration ( p <0.05). The treatment also decreased blood S -adenosylhomocysteine level (133 versus 59 nmol/L) with a borderline significance. In summary,our study shows that conventional treatment of CBS deficiency by diet and pyridoxine/betaine normalizes many but not all metabolic abnormalities associated with CBS deficiency. We propose that the finding of low plasma serine concentration in untreated CBS-deficient patients merits further exploration since supplementation with serine might be a novel and safe component of treatment of homocystinuria. PMID- 14739682 TI - Niemann-Pick disease: sixteen-year follow-up of allogeneic bone marrow transplantation in a type B variant. AB - Allogenic bone marrow transplantation (BMT) was carried out on a 3-year-old white caucasian girl with Niemann-Pick disease (NPD) type B. The donor was her unaffected brother. The patient was neurologically normal at the time of transplantation. Engraftment was based on cytogenetic studies and increased leukocyte acid sphingomyelinase (ASM) activity. However, liver biopsies taken up to 33 months post transplantation showed only a moderate reduction in stored sphingomyelin and no significant increase in ASM activity. The post transplantation period was complicated by severe graft-versus-host disease and a respiratory arrest. By 6 years of age, neurological involvement was observed, including bilateral cherry red spots. The proband is now severely mentally and physically disabled. Liver cirrhosis has continued to progress despite the BMT, and haematemesis due to portal hypertension occurred at 17 years of age. However, pulmonary infiltration regressed after BMT and there has been no clinical evidence of pulmonary insufficiency. PMID- 14739683 TI - Hearing loss in Fabry disease: the effect of agalsidase alfa replacement therapy. AB - The aim of this study was to describe the nature and prevalence of hearing loss in Fabry disease (McKusick 301500), a rare X-linked lysosomal storage disorder, and its response to enzyme replacement therapy with agalsidase alfa. Fifteen hemizygous male Fabry patients (aged 25-49 years) were randomized to receive placebo or enzyme replacement therapy for 6 months; all have received open-label enzyme replacement therapy for an additional 24 months thus far. Pure-tone audiometry, impedance audiometry and otoacoustic emission testing were performed at 0 (baseline), 6, 18 and 30 months. Four patients (27%) had bilateral and 7 (47%) had unilateral high-frequency sensorineural hearing loss (SNHL). Two (13%) had unilateral middle ear effusions with conductive losses persisting beyond 6 months. Only 3 (20%) had normal hearing. High-frequency SNHL deteriorated over the first 6 months in both placebo and active treatment groups by a median 4.3 dB ( p =0.002, Wilcoxon matched pairs). This hearing loss subsequently improved above baseline by 2.1 dB at 18 months ( p =0.02) and by 4.9 dB at 30 months ( p =0.004). In conclusion, significant hearing loss, usually high-frequency SNHL, is a common manifestation of Fabry disease in adults. alpha-Galactosidase A replacement therapy with agalsidase alfa appears to reverse the hearing deterioration in these patients. This improvement is gradual, however, suggesting the need for long-term enzyme replacement therapy. PMID- 14739684 TI - Analysis of organic acids after incubation with (16-2H3)palmitic acid in fibroblasts from patients with mitochondrial beta-oxidation defects. AB - The analysis of acylcarnitines as products of incubation of intact fibroblasts with isotope-labelled precursors, usually (16-(2)H(3))hexadecanoic acid, is an advanced in vitro method for the study of mitochondrial beta-oxidation defects. We propose a technique for the measurement of the organic acid intermediates after hydrolysis of the acylcarnitines using electron-impact gas chromatography mass spectrometry. For some mitochondrial beta-oxidation deficiencies, the characteristic profile enables us to approach the diagnosis with clear differentiation. PMID- 14739685 TI - Markedly elevated serum biotinidase activity may indicate glycogen storage disease type Ia. AB - We report two children who presented with symptoms suggestive of biotinidase deficiency. Rather than deficiency, markedly elevated serum biotinidase activities were found. Based upon literature reports of elevated biotinidase activities in children with glycogen storage disease (GSD) type Ia, we considered the latter in our differential diagnosis and subsequently confirmed GSD type Ia in both patients by enzymatic testing. GSD type Ia should be considered in children with markedly elevated serum biotinidase activity. PMID- 14739686 TI - Vaginal spasm, pain, and behavior: an empirical investigation of the diagnosis of vaginismus. AB - This study investigated the roles of vaginal spasm, pain, and behavior in vaginismus and the ability of psychologists, gynecologists, and physical therapists to agree on a diagnosis of vaginismus. Eighty-seven women, matched on age, relationship status, and parity, were assigned to one of three groups: vaginismus, dyspareunia resulting from vulvar vestibulitis syndrome (VVS), and no pain with intercourse. Diagnostic agreement was poor for vaginismus; vaginal spasm and pain measures did not differentiate between women in the vaginismus and dyspareunia/VVS groups; however, women in the vaginismus group demonstrated significantly higher vaginal/pelvic muscle tone and lower muscle strength. Women in the vaginismus group also displayed a significantly higher frequency of defensive/avoidant distress behaviors during pelvic examinations and recalled past attempts at intercourse with more affective distress. These data suggest that the spasm-based definition of vaginismus is not adequate as a diagnostic marker for vaginismus. Pain and fear of pain, pelvic floor dysfunction, and behavioral avoidance need to be included in a multidimensional reconceptualization of vaginismus. PMID- 14739687 TI - The psychological and interpersonal relationship scales: assessing psychological and relationship outcomes associated with erectile dysfunction and its treatment. AB - Erectile dysfunction (ED) is associated with complex psychological and interpersonal issues. A new measure of treatment outcome, the Psychological and Interpersonal Relationship Scales (PAIRS), was developed to evaluate the broader psychological and interpersonal outcomes associated with erectile dysfunction and its treatment. Initial items were generated based on literature review, focus groups and market research, interviews with patients and partners, and consultation with expert clinicians. Domains of Sexual Self-Confidence, Spontaneity, and Time Concerns were identified and subsequently confirmed by factor analysis. A series of validation studies was performed with four separate samples, including assessment of internal consistency and test-retest reliability, convergent and discriminant validity, and treatment responsiveness. Samples for these studies included men recruited from clinical trials for ED in several countries ( N =413) and a community sample ( N =801). Findings from these studies indicate that PAIRS demonstrates adequate psychometric properties and appears well suited for use in further clinical studies of treatments for ED. It provides a broader assessment of treatment outcome than current measures of erectile function. PMID- 14739688 TI - The effects of hysterectomy on sexual arousal in women with a history of benign uterine fibroids. AB - Research indicates hysterectomy surgery may adversely affect the pelvic autonomic nerves and autonomic mechanisms are integral to the sexual arousal response in women. This study explored the possibility that women who undergo hysterectomy may experience an impaired vasocongestive response to erotic stimulation. Thirty two women with a history of benign uterine fibroids who had ( n = 15) or had not ( n = 17) undergone hysterectomy participated in two experimental sessions in which self-report and physiological (vaginal pulse amplitude; VPA) sexual responses were recorded during an erotic film presentation. In one of the sessions, the women exercised on a treadmill for 20 min prior to viewing the erotic films as a means inducing autonomic arousal. Exercise significantly increased VPA but not subjective sexual responses in both groups of women. VPA responses were marginally higher among the fibroid than hysterectomy group in the no-exercise condition. The hypothesis that physiological sexual arousal may be impaired with hysterectomy surgery was only partially supported. PMID- 14739689 TI - Classical conditioning of sexual arousal in women and men: effects of varying awareness and biological relevance of the conditioned stimulus. AB - Classical conditioning of sexual arousal has previously been demonstrated in human males but not in females. This study explored the role of classical (Pavlovian) conditioning in the activation of genital sexual arousal in both women and men, and assessed the effects of varying conditioned stimulus (CS) duration (subliminal/conscious) and relevance (sexually relevant/irrelevant). Twenty-seven female and 29 male participants received either subliminal or conscious presentations of a photograph of either a sexually relevant (abdomen of the opposite sex) or irrelevant (gun) CS+, which was followed by the unconditioned stimulus (US-erotic film clip). A CS-, a stimulus not paired with the US, was also included in the 11 conditioning trials. Ten participants were assigned to a control group that received unpaired presentations of the CS+, CS-, and the US. Both women and men showed more evidence of conditioning to the abdomen than to the gun when the CS was presented subliminally. When consciously perceived CSs were used, however, gender differences emerged. Men again showed the expected cue-to-consequence specificity but women showed the opposite effect, that is, conditioned arousal to the sexually irrelevant rather than to the relevant CS. The latter finding may be due to increased autonomic nervous system arousal associated with the irrelevant CS (gun). Skin conductance responses indicated more general arousal to the gun than to the male abdomen in women. This is the first study to compare the effects of a subliminal and conscious CS and to find classical conditioning of sexual arousal in women. PMID- 14739690 TI - The association of sexual dysfunction and substance use among a community epidemiological sample. AB - This study examines the prevalence of DSM-III sexual dysfunctions and their association with comorbid drug and alcohol use in a community epidemiologic sample. The data for these analyses are based on the Epidemiological Catchment Area Project, a multistage probability study of the incidence and prevalence of psychiatric disorders in the general population conducted in 1981-83. Only the sample of 3,004 adult community residents in the St. Louis area was queried on DSM-III sexual dysfunctions of inhibited orgasm, functional dyspareunia (painful sex), inhibited sexual excitement (i.e., lack of erection/arousal), and inhibited sexual desire. There was a prevalence rate of 11% for inhibited orgasm, 13% for painful sex, 5% for inhibited sexual excitement, 7% for inhibited sexual desire, and 26% for any of these sexual dysfunctions (14% for men and 33% for women). The prevalence of qualifying lifetime substance use among the population was 37%, with males meeting more drug and alcohol use criteria than females. After controlling for demographics, health status variables, and psychiatric comorbidity (depression disorder, generalized anxiety disorder, antisocial personality disorder, and residual disorders), inhibited orgasm was associated with marijuana and alcohol use. Painful sex was associated with illicit drug use and marijuana use. Inhibited sexual excitement was more likely among illicit drug users. Inhibited sexual desire was not associated with drug or alcohol use. PMID- 14739691 TI - Complex demodulation of cardiac interbeat intervals: increased cardiac sympathovagal interaction during human sexual activity. AB - In this study, we analyzed heart rate variability during sexual activity using complex demodulation, which is a useful technique to study continuous changes in different frequency bands over short periods of time. Complex demodulation has the advantage of having higher time resolution compared to spectral analysis. We quantified the low frequency (LF: 0.04-0.15 Hz) and high frequency (HF: 0.15-0.5 Hz) amplitudes, and the LF/HF ratio before, during, and after the occurrence of orgasm. LF power is dually mediated by cardiac sympathetic and parasympathetic systems while HF power is mainly influenced by cardiac vagal function. We found a significant increase of cardiac sympathovagal interaction (LF/HF ratios) during the phase of orgasm. These findings are discussed in light of normal sexual physiology and the effect of sex on cardiac autonomic function in patients with heart disease, such as myocardial infarction. PMID- 14739693 TI - Mapping multiple quantitative trait Loci for ordinal traits. AB - Many complex traits in humans and other organisms show ordinal phenotypic variation but do not follow a simple Mendelian pattern of inheritance. These ordinal traits are presumably determined by many factors, including genetic and environmental components. Several statistical approaches to mapping quantitative trait loci (QTL) for such traits have been developed based on a single-QTL model. However, statistical methods for mapping multiple QTL are not well studied as continuous traits. In this paper, we propose a Bayesian method implemented via the Markov chain Monte Carlo (MCMC) algorithm to map multiple QTL for ordinal traits in experimental crosses. We model the ordinal traits under the multiple threshold model, which assumes a latent continuous variable underlying the ordinal phenotypes. The ordinal phenotype and the latent continuous variable are linked through some fixed but unknown thresholds. We adopt a standardized threshold model, which has several attractive features. An efficient sampling scheme is developed to jointly generate the threshold values and the values of latent variable. With the simulated latent variable, the posterior distributions of other unknowns, for example, the number, locations, genetic effects, and genotypes of QTL, can be computed using existing algorithms for normally distributed traits. To this end, we provide a unified approach to mapping multiple QTL for continuous, binary, and ordinal traits. Utility and flexibility of the method are demonstrated using simulated data. PMID- 14739694 TI - Using the Mplus computer program to estimate models for continuous and categorical data from twins. AB - Historically, the focus of behavior genetic research was to obtain estimates of the sources of familial resemblance for a single phenotype. Current research strategies have moved beyond heritability estimates to the search for physiological and behavioral mechanisms by which genetic risk is translated into individual differences in behavior and disease liability. Such research questions often require multivariate designs and complex analytic models, including the analysis of continuous and categorical dependent variables within the same model. Recent advances in computer software for categorical data analysis have increased the tools available for researchers in behavior genetics. This paper describes how to use the Mplus software program (Muthen and Muthen, 1998, 2002) for the analysis of data obtained from twins. Example analyses include two- and five group twin models for univariate and bivariate continuous and categorical variables. Data on alcoholism and age at first drink drawn from the Virginia Adult Twin Study of Psychiatric and Substance Use Disorders are used to illustrate how Mplus can be used to analyze multiple-category variables, recode and transform variables, select subgroups for analysis, handle subjects with incomplete data, include constraints to ensure non-negative loadings, include model covariates, model sex differences, and test alternative hypotheses about mediation of genetic risk by measured variables. PMID- 14739695 TI - A genetic investigation of the covariation among inspection time, choice reaction time, and IQ subtest scores. AB - Information processing speed, as measured by elementary cognitive tasks, is correlated with higher order cognitive ability so that increased speed relates to improved cognitive performance. The question of whether the genetic variation in Inspection Time (IT) and Choice Reaction Time (CRT) is associated with IQ through a unitary factor was addressed in this multivariate genetic study of IT, CRT, and IQ subtest scores. The sample included 184 MZ and 206 DZ twin pairs with a mean age of 16.2 years (range 15-18 years). They were administered a visual (pi figure) IT task, a two-choice RT task, five computerized subtests of the Multidimensional Aptitude Battery, and the digit symbol substitution subtest from the WAIS-R. The data supported a factor model comprising a general, three group (verbal ability, visuospatial ability, broad speediness), and specific genetic factor structure, a shared environmental factor influencing all tests but IT, plus unique environmental factors that were largely specific to individual measures. The general genetic factor displayed factor loadings ranging between 0.35 and 0.66 for the IQ subtests, with IT and CRT loadings of -0.47 and -0.24, respectively. Results indicate that a unitary factor is insufficient to describe the entire relationship between cognitive speed measures and all IQ subtests, with independent genetic effects explaining further covariation between processing speed (especially CRT) and Digit Symbol. PMID- 14739696 TI - Familial aggregation patterns in mathematical ability. AB - Mathematical talent is an asset in modern society both at an individual and a societal level. Environmental factors such as quality of mathematics education undoubtedly affect an individual's performance, and there is some evidence that genetic factors also may play a role. The current study was performed to investigate the feasibility of undertaking genetics studies on mathematical ability. Because the etiology of low ability in mathematics is likely to be multifactorial and heterogeneous, we evaluated families ascertained through a proband with high mathematical performance in grade 7 on the SAT to eliminate, to some degree, adverse environmental factors. Families of sex-matched probands, selected for high verbal performance on the SAT, served as the comparison group. We evaluated a number of proxy measures for their usefulness in the study of clustering of mathematical talent. Given the difficulty of testing mathematics performance across developmental ages, especially with the added complexity of decreasing exposure to formal mathematics concepts post schooling, we also devised a semiquantitative scale that incorporated educational, occupational, and avocational information as a surrogate for an academic mathematics measure. Whereas several proxy measures showed no evidence of a genetic basis, we found that the semiquantitative scale of mathematical talent showed strong evidence of a genetic basis, with a differential response as a function of the performance measure used to select the proband. This observation suggests that there may be a genetic basis to specific mathematical talent, and that specific, as opposed to proxy, investigative measures that are designed to measure such talent in family members could be of benefit for this purpose. PMID- 14739697 TI - A twin study of competence and behavioral/emotional problems among adolescents in taiwan. AB - This work reports on a study to evaluate the relative contributions of genetic and environmental factors to both competence scales and behavioral/emotional syndromes as assessed by the Child Behavior Checklist (CBCL). A total of 279 pairs of twins and same-sex sib-pairs aged 12-16 years were recruited from 51 junior high schools in Taipei City, Taiwan. Twins' zygosity was determined by a combination of DNA typing and physical similarity. The Mx program was used to estimate parameters for a full model that contains effects from sex-specific additive genes, shared environment, and nonshared environment for the majority of the scales. The shared environment in the full model was replaced with nonadditive genetic factors for some scales when indicated. All girls' competence and behavioral/emotional syndromes exhibited a substantial heritability (h2 > 0.4), except for Social Competence and Withdrawn. For boys, though the heritability was also >0.4 for some scales (Social and School Competence, Thought Problems, Attention Problems, Delinquent Behavior, and Total Behavior Problems), environmental influences, especially shared environment, were predominant for most of the scales (10 out of 15 scales). Genetic factors are important for explaining adolescent behavioral problems, especially for girls, while shared environmental influences cannot be ignored for boys. Gender differences in heritability exist for various CBCL-based competence and behavioral/emotional problems. PMID- 14739698 TI - Are personality disorders psychological manifestations of executive function deficits? Bivariate heritability evidence from a twin study. AB - This study tested whether personality disorders may be the psychological manifestations of executive function deficits by examining their bivariate heritability in a community sample of 314 twins (ages 5-17 years; M age = 9.7; 96 monozygotic pairs and 61 dizygotic pairs). The parents of the twins completed the Coolidge Personality and Neuropsychological Inventory (Coolidge, 1998; Coolidge et al., 2002). Heritability was estimated by structural equation modeling. Executive function deficits and personality disorders were significantly heritable (executive function deficits,.77; 11 out of 12 personality disorders, median =.69). The proportion of the observed correlation attributable to heritable factors or bivariate heritability between executive function deficits and the personality disorder scales ranged from.27 for schizoid to.64 for histrionic. These findings may provide some insight as to why individuals diagnosed with specific personality disorders frequently exhibit chronic difficulties with everyday decisions, selective attention and inhibition, judgments, choices, planning, and flexibility. PMID- 14739699 TI - Association between dependent smoking and a polymorphism in the tyrosine hydroxylase gene in a prospective population-based study of adolescent health. AB - This study reports pilot data on an association between tobacco dependence and a five-allele tetranucleotide repeat polymorphism in the first intron of the tyrosine hydroxylase (TH) gene. One hundred and twenty-six Australian adolescents who had participated in the Health in Transition Study (1993-1997), and who showed patterns of either dependent or nondependent smoking across four waves of data collection, consented to participation in the pilot study. The smoking status of those recruited was confirmed using a telephone-administered drug use questionnaire during 2000. Tobacco dependence was defined as smoking more than 6 days per week and more than 10 cigarettes per day during wave 5 (year 2000) and at least one prior wave ( n = 58). A second, more stringent phenotype included smoking within an hour of waking ( n = 37). The control group comprised adolescents who had used tobacco but had remained low-level social smokers across each wave of data ( n = 56). DNA was collected using a mouthwash procedure. Using the more strictly defined tobacco dependence phenotype, and after adjusting for sex, a significant protective association was found between the K4 allele and tobacco dependence (OR 0.27, 95% confidence interval [CI] 0.09, 0.82). No association was found using the liberal criteria of tobacco dependence (OR 0.51, 95% confidence interval [CI] 0.23, 1.2). These preliminary results replicate a previous association between tobacco use and the K4 allele of the TH gene (Lerman et al., 1997). The potential significance of including time to first cigarette in definitions of tobacco dependence and the possible role that these TH variants might play in tobacco dependence are discussed. PMID- 14739700 TI - Identification of quantitative trait Loci for anxiety and locomotion phenotypes in rat recombinant inbred strains. AB - Anxiety disorders are phenotypically complex and may involve multiple genetic influences on many neurotransmitter systems. Rodent tests used to investigate genetic influences on anxiety-like phenotypes have face and predictive validity as models for anxiety in humans. If multiple genes contribute additively to a trait, the trait will be continuously distributed and be amenable to detection of associations between allelic variation at specific chromosomal loci and the phenotypes being studied via quantitative trait loci (QTL) mapping. The elevated plus-maze test provides quantitative measures of both anxiety-like and locomotion phenotypes. Using this test, we assessed four phenotypes in a set of 22 rat recombinant inbred (RI) strains derived from Brown Norway (BN.Lx /Cub) and Spontaneously Hypertensive rat (SHR/Ola) progenitors. QTL analyses were used to determine whether allelic variation at specific chromosomal loci contribute significantly to RI strain-dependent variance in each phenotype. Significant QTL for an anxiety phenotype were found on chromosomes 2, 5, 6, and 7. For a phenotype reflecting both anxiety and locomotion, QTL were found on chromosomes 2, 7, and 8, while for a locomotion phenotype, significant QTL were found on chromosomes 3 and 18. PMID- 14739701 TI - Operant conditioning in the Ts65Dn mouse: learning. AB - Ts65Dn and littermate controls were trained to respond (nose-poke) under operant schedules of reinforcement. A small difference was observed in the initial operant training of the Ts65Dn mouse that disappeared with training under a fixed ratio 15 schedule of milk presentation. No difference was observed in a position reversal task in which mice initially trained to respond upon one photocell had to learn to respond on a previously inactive photocell. Under an incremental repeated acquisition of behavioral chains schedule, no difference was observed when one or two responses were required to complete the sequence. However, when three or four responses were required to complete the sequence, a marked deficit was observed in the Ts65Dn mice. These results show that both Ts65Dn and littermate controls can be trained to respond under operant schedules of reinforcement, but that Ts65Dn mice have a learning deficit that is correlated with task difficulty. PMID- 14739702 TI - Major growth QTLs in fowl are related to fearful behavior: possible genetic links between fear responses and production traits in a red junglefowl x white leghorn intercross. AB - The aim of this work was to study fear responses and their relation to production traits in red junglefowl ( Gallus gallus spp.), White Leghorn ( Gallus domesticus ), and their F2-progeny. Quantitative trait locus (QTL) analyses were performed for behavioral traits to gain information about possible genetic links between fear-related behaviors and production. Four behavioral tests were performed that induce different levels of acute fear (open field [OF], exposure to a novel object, tonic immobility, and restraint). Production traits, that is, egg production, sexual maturity (in females), food intake, and growth, were measured individually. A genome scan using 105 microsatellite markers was carried out to identify QTLs controlling the traits studied. In the OF and novel object tests (NO), Leghorns showed less fear behavior than junglefowl, whereas junglefowl behaved less fearfully in the tonic immobility test (TI) and were more active in the restraint test. In the F2 progeny, only weak phenotypic associations were found between production traits and fear behavior. A significant QTL for TI duration was found on chromosome 1 that coincided with a QTL for egg weight and growth in the same animals. Another QTL for NO in males coincided with another major growth QTL. These two known growth QTLs affected a wide range of reactions in different tests. Several other significant and suggestive QTLs for behavioral traits related to fear were found. These QTLs did not coincide with QTLs for production traits, indicating that these fear variables may not be genetically linked to the production traits we measured here. The results show that loci affecting important production traits are located in the same chromosomal region as loci affecting different fear-related behaviors. PMID- 14739703 TI - Number of arginine-vasopressin neurons in the suprachiasmatic nuclei is not related to level or circadian characteristics of wheel-running activity in house mice. AB - House mouse lines bidirectionally selected for nest-building behavior show a correlation between number of AVP cells in the suprachiasmatic nuclei (SCN), the master circadian clock in mammals, and level of nest-building behavior as well as a correlation between wheel-running activity and SCN AVP content. Similar genetic correlations between wheel-running activity and nest-building behavior have been reported in house mouse lines selected for increased voluntary wheel-running behavior. These similarities in genetic correlation structure in independently selected mouse lines allowed us to test whether AVP in the SCN and wheel running activity are truly correlated traits under identical testing procedures. In the mouse lines selected for voluntary wheel-running, no difference was found between the lines selected for high levels of voluntary wheel-running and randomly-bred control lines in the number of AVP immunoreactive neurons in the SCN ( F1,6 = 0.09, NS; replicate line effect: F1,22 = 0.05, NS). This finding was confirmed at the level of individual variation, which revealed no relationship between number of AVP neurons in the SCN and total daily activity ( R = -0.086, NS, n = 24), or circadian organization (i.e., the chi-squared periodogram waveform amplitude; R = -0.071, NS). Therefore our data do not support the hypothesis that differences in activity level and the circadian expression of activity in young adult mice are related to differences in the number of AVP-immunoreactive cells in the SCN. PMID- 14739704 TI - Effect of increasing age on tissue dolichol levels in ad libitum fed and food restricted rats. AB - In order to test the hypothesis that the ageing-related alteration in membrane lipids might reflect the biological age of rodents, we studied the effects of age in ad libitum fed (AL) and food-restricted (FR) male Sprague-Dawley rats on the levels of dolichol in different organs involved [liver (L) and kidney (K)] or not involved [brain (B), sciatic nerve (SN), heart (H), soleus (S) and extensor digitorum longus (EDL) muscles] in dolichol excretion. At the given age, tissue dolichol was extracted and assayed by HPLC procedure. Results show that the levels of dolichol were significantly different in different tissues and increased dramatically with increasing age. The anti-ageing FR regimen had significant preventive effects on dolichol accumulation in the excretory organs. The effect of FR on the liver was much bigger than that of kidney. The effect of FR retarding dolichol accumulation in the liver co-varied with the effects of FR on longevity. In conclusion, these data show that the quantity of dolichol in the hepatic tissue might be used as a marker of the biological age of the animal. PMID- 14739705 TI - The fate of dolichol in rat cells and tissues. AB - Dolichol (D) levels increase dramatically in older tissue. A better understanding of the fate of cell D and exchange between tissues could be essential for understanding the mechanism of the abnormal accumulation. The fate of red blood cell D was investigated by the use of phenylhydrazine-induced hyperhaemolysis. The effect of atrophy on D tissue levels was studied in the perineal muscles of castrated rats. Influence of D transportation between tissues on the levels of D was studied by the use of age-mismatched heterotopic transplantation of D-rich hearts from older (22 months old) donor rats in younger (3 months old) D-poor syngenic recipients. Increased red blood cell destruction by splenic macrophages did not cause accumulation but rather a significant depletion of the D content of the spleen. The shrinkage of tissues by endocrine or disuse atrophy did not affect the D content of muscle, where D concentration increased. No significant net redistribution of D was observed from the transplanted older heart to liver and tissues of younger recipients. In conclusion, phagocytosis appears to be the only process resulting in the disposal of tissue D. PMID- 14739706 TI - New perspectives for (S)-dolichol and (S)- nor dolichol synthesis and biological functions. AB - A procedure is described for the preparation of (S)-dolichol and (S)- nor dolichol starting from the polyprenyl fraction extracted from Gingko biloba integer or exhausted leaves. The procedure appears extremely valuable in obtaining the two chiral isoprenoid compounds in good chemical yields and retention of a high degree of enantiomeric excess. Also, the (S)- nor dolichol represents a good chiral precursor for the preparation of (14)C-labelled (S) dolichol to be used in biological investigations into the (S)-dolichol catabolism in the functional living cell. Furthermore, the possible role of (S)-dolichol as a free radical scavenger in the cell membrane was preliminarily evaluated by means of a (1)H-NMR analytical method. Apparently, experimental results substantiate this hypothesis. PMID- 14739707 TI - The effect of carbon tetrachloride and ultraviolet radiation on dolichol levels in liver cells isolated from 3- and 24-month-old male Sprague-Dawley rats. AB - Dolichol (D) is a long-chain polyprenoid broadly distributed in the cell membranes, possibly endowed with a free-radical scavenging activity, whose concentration in tissues increases with increasing age. No enzyme pathway for D degradation has been discovered. In order to test the hypothesis that D might undergo a non-enzymatic free-radical mediated decomposition the effects of a xenobiotic agent (carbon tetrachloride, CCl(4)) and ultraviolet-B (UV-B) radiation on D levels were studied in liver cells isolated from male ad libitum fed Sprague-Dawley rats aged 3 or 24 months. Liver cells (90 mg/ml) were incubated in sealed flasks (6 ml cell suspension each) for 0, 5, 10 and 20 min after the addition of 25, 50 or 200 microl CCl(4) in the central well. 50 ml of a 6 mg/ml liver cell suspension were poured in a 120 cm(2) Petri dish and the sediment liver cell monolayer was exposed to UVB radiation for 0, 5, 10, 20 and 40 min. At the given time, cells were taken and D was extracted and assayed by the HPLC procedure. D levels were remarkably higher in older than in younger cells as expected ( P < 0.001). Treatment with CCl(4) and UVB caused a highly significant decrease in D ( P < 0.001) whose percentage was larger in younger than in older cells. The conclusions are that free-radicals generated either by chemical or by physical agents cause a very rapid depletion of D in liver cells, and that the effect of the free radical attack on D decomposition may be lower percentage wise in older than in younger cells, which might account at least in part for the accumulation of D in older tissues. PMID- 14739708 TI - Age-related changes of isoprenoid biosynthesis in rat liver and brain. AB - The physiological role of dolichol is not yet known but its accumulation in several tissues has been extensively reported in various physiological states or pathological conditions. Increased dolichol concentration in mammalian tissues during ageing has been also reported; in particular, we have previously indicated dolichol accumulation in liver as a new biomarker of ageing. However, the mechanism and the role of this accumulation is unknown. The aim of this work was to study the mechanism of the age-dependent dolichol accumulation analysing, in the liver and in the brain, the activity of the rate-limiting enzyme of isoprenoid biosynthesis, the 3-hydroxy 3-methylglutaryl CoA reductase, the dolichol and cholesterol synthesis on aged rats both fed ad libitum and caloric restricted. Furthermore, the dolichol and cholesterol levels in the plasma were assayed. The data shows that during ageing, the tissue dolichol accumulation is connected with the increase of 3-hydroxy 3-methylglutaryl CoA reductase activity and only in liver affected by diet restriction. In addition the aged rats maintain the capability to regulate their tissue cholesterol content by modifying cholesterol delivery into the blood. The amount of the 3-hydroxy 3-methylglutaryl CoA reductase enzyme detectable in liver and brain by Western blot analysis does not show significant changes during ageing. The presented data show that the accumulation of dolichol is related to the loss of enzymatic regulation characteristic of ageing. In fact, a higher mevalonate availability deriving from an increased expressed activity of HMGCoA-R could cause an increased production of dolichol. PMID- 14739710 TI - Reproduction and survival in Mediterranean fruit flies: a "protein and energy" free radical model of aging. AB - We propose a "protein and energy" free radical model of aging that predicts patterns of survival and fertility curves for Mediterranean fruit flies. Mathematical and simulation models of individual physiological processes were constructed in terms of stochastic differential equations. The free radical theory of aging was used as a basis for construction of the model and was extended to describe the dynamics of the protein and energy balance in the functioning organism, including the concept of a maximum capacity for energy production. We fit the model to observed patterns of survival and fertility in flies given the basic diet schemes used in respective experiments. Age patterns of fertility and survival were predicted under various diet scenarios using the model, revealing that the predictions of the model are in good agreement with experimental data. PMID- 14739709 TI - Dolichol: a solar filter with UV-absorbing properties which can be photoenhanced. AB - Dolichol, the polyisoprenoid lipid found in all eukaryotic cells and suggested to represent a biomarker of aging, is inserted into cell membranes, also in tissues exposed to light such as the skin. A general question about its physiological role is whether dolichol may play the role of a natural barrier for the noxious components of solar radiation. In order to clarify this point, we established that dolichol is a component of human sebum and we performed an " in vitro " study of the effects of UV radiation on the spectral properties of dolichol in isopropanol. Our data clearly show that, following UV irradiation, the optical absorption spectrum of dolichol undergoes remarkable modifications below 400 nm: a significant, strongly dose-dependent, increase of the optical density around 320 nm and a minor, very slightly dose-dependent, raise of the absorbance at 250 nm. On the contrary, UV irradiation causes only minor changes in HPLC profiles and the formation of photooxidative products can be considered negligible in our experimental conditions. These results suggest that dolichol can be considered an innate, unusually efficient and promising UV screen for skin protection. PMID- 14739711 TI - Errors, mitochondrial dysfunction and ageing. AB - Experiments in yeast have shown that increasing the accuracy of mitochondrial ribosomes increases cellular lifespan, which suggests that mitochondrial ribosomal inaccuracy could be a potential source age-related mitochondrial dysfunction. It is argued that experiments used to test the error-catastrophe theory of ageing could therefore be invalid because only proteins synthesized on 80S cytoplasmic ribosomes were analyzed. It is speculated that in elderly mammals, when growth and mitochondrial replacement rates decline, the increased organelle dwell-time allows manifestation of any loss of mito-ribosomal accuracy. The resultant aberrant polypeptides could then overload the Lon protease, which also degrades oxidatively damaged polypeptides, and thereby contribute to the accumulation of damaged macromolecules, mitochondrial dysfunction and senescence. It is suggested that a search for age-related changes in the accuracy of mitochondrial ribosomes be carried out to test the validity or otherwise of these ideas. PMID- 14739712 TI - "I thought, thought, thought for four months in vain and suddenly the idea came"- an interview with Denham and Helen Harman. Interview by K. Kitani and G.O. Ivy. PMID- 14739713 TI - Significance of atrial fibrillation during acute myocardial infarction, and its current management: insights from the GUSTO-3 trial. AB - The Global Use of Strategies to Open Occluded Coronary Arteries (GUSTO)-3 atrial fibrillation (AF) substudy assessed the prognostic significance of AF during acute myocardial infarction (AMI), the use of antiarrhythmic therapies, and whether different antiarrhythmic therapies were associated with different outcomes. The timing of the onset of AF relative to other post-AMI complications was recorded in the study. Of the 13,858 patients who were in sinus rhythm at the time of enrolment into GUSTO-3, 906 (6.5%) developed AF and 12,952 did not. Worsening heart failure, hypotension, third-degree heart block, and ventricular fibrillation were independent predictors of new-onset AF. The risks of 30-day and 1-year mortality were increased among patients with AF versus patients without AF before (odds ratio [OR] 2.74, 95% confidence interval [CI] 2.56-3.34; and OR 2.93, 95% CI 2.48-3.46, respectively) and after adjustment for baseline factors and pre-AF complications (OR 1.49, 95% CI 1.17-1.89; and OR 1.64, 95% CI 1.35 2.01, respectively). A total of 1,138 patients had data available on the management of their AF, including 117 with a history of paroxysmal AF and 138 with chronic AF prior to AMI. Of these 1,138 patients, 317 (28%) received antiarrhythmic therapies: class I antiarrhythmic drugs in 12%, sotalol in 5% and amiodarone in 15%. Electrical cardioversion was attempted in 116 patients (10%). Sinus rhythm was restored in 72% of patients given class I drugs, 67% of those given sotalol, 79% of those given amiodarone, and 64% of those who underwent electrical cardioversion. After adjustment for baseline characteristics and pre AF complications, none of the specific antiarrhythmic therapies was associated with a higher chance of having sinus rhythm at discharge or before deterioration to in-hospital death. However, the use of class I antiarrhythmic drugs or sotalol was associated with lower unadjusted 30-day and 1-year mortality rates. After adjustment for baseline factors and pre-AF complications, the ORs for 30-day and 1-year mortality were 0.42 (95% CI 0.19-0.89) and 0.58 (95% CI 0.33-1.04), respectively, with class I agents, and 0.31 (95% CI 0.07-1.32) and 0.31 (95% CI 0.09-1.02), respectively, with sotalol. In contrast, there was no association between the use of amiodarone or electrical cardioversion and 30-day or 1-year mortality. New AF is often secondary to other post-AMI complications, but is in itself an independent predictor of a worse outcome. Clinical management of AF is variable, but in GUSTO-3 there was a strong trend towards lower mortality associated with the use of class I antiarrhythmic agents or sotalol. Randomized trials are needed to investigate this observation further. PMID- 14739714 TI - Rationale for the Atrial Fibrillation and Congestive Heart Failure (AF-CHF) trial. AB - Congestive heart failure and atrial fibrillation (AF) are two important and growing problems in medicine and cardiology. Both conditions often coexist and complicate each other's management. Two therapeutic strategies are available for patients with AF and congestive heart failure: the first aims at restoring and maintaining sinus rhythm, whereas the second focuses exclusively on optimizing ventricular rate. Prior studies of AF and congestive heart failure were not randomized and most were retrospective. Although some studies suggested that AF had no effect on survival, in most recent large congestive heart failure trials, AF was reported to be an independent risk factor for mortality or major morbidity. The primary objective of the Atrial Fibrillation in Congestive Heart Failure (AF-CHF) trial is to compare the two widely used treatment strategies with respect to cardiovascular mortality. AF-CHF is a prospective, multicenter trial that will randomize 1450 CHF patients with left ventricular ejection fraction (LVEF) < or =35% and a documented recent episode of atrial fibrillation to either a rhythm control or a rate control strategy. From recent trial data, we anticipate an 18.75% 2-year cardiovascular mortality in the rate control arm and a 25% event reduction in the rhythm control group. As of December 2003, 960 patients have been randomized. Enrollment is expected to be completed in September 2004 with a minimum follow-up of 2 years. PMID- 14739715 TI - Cost evaluation of rhythm control methods for atrial fibrillation: evidence from CTAF. AB - Atrial fibrillation (AF) is a highly prevalent arrhythmia that is difficult to treat and generates important health care costs. One consideration in the selection of various therapeutic options is the cost of a given treatment compared to that of alternatives. The Canadian Trial of Atrial Fibrillation (CTAF) evaluated the effectiveness of sinus rhythm maintenance with amiodarone compared to propafenone or sotalol in a prospective, randomized fashion. A subsequent CTAF substudy of the medical costs associated with amiodarone vs. propafenone/sotalol found that amiodarone decreased AF-related costs. This paper reviews the results of the CTAF cost-analysis substudy in the context of other analyses in the literature of the cost effectiveness of amiodarone in AF. The costs associated with amiodarone therapy are no greater than for other sinus rhythm maintenance drugs, and for some cost categories and some patient subgroups are likely to be less, despite amiodarone's greater therapeutic efficacy. However, additional considerations are important in evaluating the clinical place of amiodarone, including its adverse effect and pharmacokinetic profile. As well, the results of recent randomized clinical trials have highlighted the limitations of sinus rhythm maintenance as a primary therapeutic objective in AF. The decision about whether and at what point to use amiodarone in a given patient requires a careful analysis of the individual case, in terms of symptomatology during AF, the response to previous treatment regimes, and risk factors for various forms of adverse drug reactions. PMID- 14739716 TI - Azimilide for atrial fibrillation: clinical trial results and implications. AB - Azimilide dihydrochloride (or azimilide) is a class III antiarrhythmic drug currently under investigation that has been tested in atrial fibrillation in four randomized, placebo-controlled clinical trials to assess efficacy and dose range. These investigational trials showed that doses of azimilide 100 and 125 mg once daily prolonged the time to symptomatic arrhythmia recurrence in patients with a history of symptomatic atrial fibrillation, atrial flutter or both. Doses of 75 mg or less were not useful in this indication. Safety of azimilide has been examined in several different types of studies. In a large randomized clinical trial of post-infarct patients, azimilide neither increased nor decreased mortality risk. In patients with supraventricular arrhythmias, the most common adverse effects reported by patients on azimilide were approximately equal in frequency with those on placebo: headache, asthenia, infection, diarrhea and dizziness. Infrequent cases of torsade de pointes and severe neutropenia were reported in patients taking azimilide. Azimilide is an investigational antiarrhythmic drug that has shown efficacy in patients with atrial fibrillation. PMID- 14739717 TI - Does conversion and prevention of atrial fibrillation enhance survival in patients with left ventricular dysfunction? Evidence from the Danish Investigations of Arrhythmia and Mortality ON Dofetilide/(DIAMOND) study. AB - BACKGROUND: Atrial fibrillation is a common arrhythmia in patients with left ventricular dysfunction associated with increased morbidity and mortality. The present study investigated the potential of dofetilide to restore and maintain sinus rhythm in patients with left ventricular dysfunction, which might reduce mortality and hospitalizations. METHODS AND RESULTS: In the Danish Investigations of Arrhythmia and Mortality ON Dofetilide (DIAMOND) studies, 506 patients were in atrial fibrillation (AF) or atrial flutter (AFl) at baseline. Over the course of study, cardioversion occurred in 148 (59%) dofetilide- and 86 (34%) placebo treated patients. In these patients, the probability of maintaining sinus rhythm for 1 year was 79% with dofetilide versus 42% with placebo ( P < 0.001). Dofetilide had no effect on all-cause mortality, but restoration and maintenance of sinusrhythm (independent of study treatment) was associated with a significant reduction in mortality (risk ratio [RR], 0.44; 95% CI, 0.30 to 0.64; P < 0.0001). In addition, dofetilide therapy was associated with a significantly lower risk ratio versus placebo for either all-cause (RR, 0.70; 95% CI, 0.56 to 0.89; P < or = 0.005) or congestive heart failure (RR, 0.69; 95% CI, 0.51 to 0.93; P < or = 0.02) rehospitalization. CONCLUSIONS: Dofetilide is safe and increases the probability of obtaining and maintaining sinus rhythm in patients with structural heart disease. The present study suggests that restoration of sinus rhythm--on placebo or dofetilide--is associated with improved survival. PMID- 14739718 TI - Role of dofetilide in patients with atrial fibrillation. Insights from the Symptomatic Atrial Fibrillation Investigative Research on Dofetilide (SAFIRE-D) study. AB - Dofetilide is a class III antiarrhythmic drug (potassium channel blocker) that has been approved by the regulatory agencies in the United States and throughout the world to convert atrial fibrillation and maintain sinus rhythm. Therapy is initiated in-hospital during heart rhythm monitoring. Doses are selected according to the QT interval and estimated creatinine clearance. In patients with heart failure and prior myocardial infarction, dofetilide was very effective in converting atrial fibrillation and maintaining normal sinus rhythm. In patients with persistent atrial fibrillation, dofetilide compared to placebo was significantly better in converting atrial fibrillation and maintaining sinus rhythm. This was especially true for the highest dose of 500 microg twice-a-day. Caution must be used when initiating dofetilide therapy to avoid torsade de pointes ventricular tachycardia, especially in patients with heart failure, hypertrophy, bradycardia and female gender. PMID- 14739719 TI - Beta blockers improve outcome in patients with heart failure and atrial fibrillation: U.S. carvedilol study. AB - Atrial fibrillation (AF) is present in a significant number of patients with heart failure (HF) caused by left ventricular systolic dysfunction and is associated with increased morbidity and mortality. The deleterious interaction of AF and HF is mediated through a number of mechanisms including hemodynamic alterations and activation of the sympathetic nervous system. Beta-blockers have been shown to improve symptoms and survival in patients with HF. In addition, beta-blockers have been used in patients with AF, primarily for rate control. A retrospective analysis of the U.S. Carvedilol Heart Failure Trial demonstrated that carvedilol improves outcomes in the high-risk subgroup of patients with HF and concomitant AF. PMID- 14739720 TI - Beta-blocker efficacy according to heart rate and rhythm in patients with heart failure. Commentary on the Cardiac Insufficiency Bisoprolol Study II analysis. AB - Large randomized trials have demonstrated that beta-blocker treatment reduces morbidity and mortality in patients in chronic heart failure. Questions remain about the influence of individual characteristics on the magnitude of the benefit of beta-blockers in patients with heart failure including the influence of heart rate and cardiac rhythm. In the Cardiac Insufficiency Bisoprolol Study II, baseline heart rate and heart rate change over time had prognostic value but treatment with bisoprolol was associated with a benefit at all levels of baseline heart rate and additional benefit related to heart rate slowing was observed. In the subgroup of patients with atrial fibrillation, morbidity and mortality rates were similar in placebo and bisoprolol treated patients. It is possible that patients with atrial fibrillation had a higher level of sympathetic stimulation that would have required higher doses of bisoprolol to achieve a similar level of beta-blockade. Alternatively, the failure to observe improved outcome in the subgroup with atrial fibrillation may have been due to chance. However, because this finding was not observed in other large trials, and because there was no clear explanation, it should not be concluded that patients with chronic heart failure and atrial fibrillation do not benefit from beta-blockade. PMID- 14739721 TI - Role of beta-blocker therapy in heart failure and atrial fibrillation. AB - Heart failure is a serious disorder associated with substantial morbidity and mortality. Approximately 15-30% patients with systolic heart failure are in atrial fibrillation and the proportion increases with severity of heart failure. Patients with heart failure and atrial fibrillation have worse outcome than those in sinus rhythm. Beta-blockers, together with angiotensin-converting enzymes inhibitors, are the standard therapy in patients with chronic heart failure. Retrospective studies have suggested that despite the improvement in left ventricular systolic function after treatment with beta-blockers, the exercise capacity and symptoms in those heart failure patients with atrial fibrillation was not improved as much as those in sinus rhythm. Moreover, the use of bisoprolol in the Cardiac Insufficiency Bisoprolol Study II, unlike those in sinus rhythm, failed to produce any survival benefit in patients with poor systolic function and atrial fibrillation. It seems that those patients with heart failure and atrial fibrillation may have different response to beta-blocker therapy. Prospective trials to clarify the impact of beta-blocker therapy and the optimal therapeutic strategy in this high-risk group of patients are warranted. PMID- 14739722 TI - Angiotensin receptor blocker as adjunctive therapy for rhythm control in atrial fibrillation: results of the irbesartan-amiodarone trial. AB - Atrial fibrillation (AF) is a common arrhythmia associated with increased risk of stroke and mortality. The early appearance of electrical remodeling is followed by structural remodeling of the atrial tissue. Direct current cardioversion of persistent AF is the most effective treatment for the restoration of sinus rhythm, but it is hampered by a high percentage of recurrences. Recurrences may be the consequence of both electrical and structural remodeling. A study on the use of irbesartan to maintain sinus rhythm in patients with long-lasting persistent AF showed that this angiotensin II receptor blocker combined with amiodarone prolonged sinus rhythm after cardioversion. Irbesartan may have antifibrotic effects due not only to the ability to diminish the synthesis of collagen type I molecules but also to its capacity to stimulate the degradation of collagen type I fibers, as has been demonstrated with losartan, another angiotensin II receptor blocker. This suggests that efforts to reduce the structural changes that occur during AF may be more useful in preventing recurrences than efforts designed to minimize the electrical changes alone. The AFFIRM trial compared two approaches to the treatment of AF: cardioversion with antiarrhythmic drugs to maintain sinus rhythm and the use of rate-controlling drugs. The results show that management of AF with the rhythm-control strategy offers no survival advantage over the rate-control strategy. However, non antiarrhythmic drugs to prevent recurrences, like irbesartan, were not controlled and amiodarone was used in a low percentage of the patients. The treatment strategies proposed in both AFFIRM and RACE, in our opinion, may not be the optimal. The modern clinical approach to AF involves an early intervention to restore sinus rhythm, therefore preventing atrial remodeling. The pretreatment of patients with AF who undergo electrical cardioversion is very important and will be the subject for continuous improvement. PMID- 14739723 TI - Conducting randomized trials in the electrophysiology laboratory: lessons from a randomized comparison of recording methods during pulmonary vein isolation by segmental ostial ablation. AB - Accurate identification of pulmonary vein (PV) potentials during segmental ostial ablation for PV isolation is very important to completely isolate the PVs and also to avoid unnecessary applications of radiofrequency energy. A prior post hoc analysis of unipolar and bipolar electrograms recorded from successful and unsuccessful ablation sites demonstrated that unipolar electrograms recorded at successful sites were more likely to have a rapid intrinsic deflection, larger amplitude, and earlier activation than electrograms recorded at unsuccessful sites. Moreover, unipolar electrograms recorded from the ablation and circular ostial mapping catheters were almost identical at successful sites. Based on these observations, a prospective, randomized study was conducted to test whether unipolar and bipolar electrograms would facilitate the ablation procedure when compared to bipolar electrograms alone during PV isolation in patients with atrial fibrillation (AF). In 44 consecutive patients with paroxysmal AF, 114 PVs were randomized to segmental ostial ablation guided by unipolar and bipolar electrograms (61) or by bipolar electrograms only (53). Segmental ostial ablation guided by unipolar and bipolar electrograms was associated with a approximately 20-30% decrease in the procedure and fluoroscopy times necessary for isolation of a PV and also in the duration of radiofrequency energy application required for complete isolation of a PV. Although the sample size was not sufficient to detect a 5% change, the success rate for complete electrical isolation and the risk of symptomatic PV stenosis were similar between the 2 groups. Online analysis of unipolar electrograms facilitated the PV isolation procedure and was incremental to the analysis of bipolar electrograms alone. However, because segmental ostial ablation has only modest efficacy in achieving long-term freedom from recurrent AF, alternative ablation strategies that may or may not target PVs will eventually evolve. The role of unipolar electrograms in these new methods remains to be determined. PMID- 14739724 TI - Left versus bi-atrial intraoperative saline-irrigated radiofrequency modified maze procedure for atrial fibrillation. AB - BACKGROUND: This study was conducted to evaluate the effectiveness of the saline irrigated radiofrequency modified maze operation for the treatment of chronic atrial fibrillation (AF) and compare the results of the left and bi-atrial procedures. MATERIAL AND METHOD: During a period of two years, 105 patients with chronic AF having concomitant cardiac surgery underwent the procedure. Patients underwent either a bi-atrial ( n = 48) or left atrial ( n = 57) maze procedure. The first twenty patients underwent a bi-atrial maze procedure regardless of the pathology. In the following patients we adopted the bi-atrial approach in patients with a history of atrial flutter and where the right atrium has to be opened. Otherwise the procedure is restricted to the left atrial side. Mean age was 52 +/- 11 years in bi-atrial group and 54 +/- 9 years in left atrial group. RESULTS: Three patients died early postoperatively (2.9%). There were 4 revisions for bleeding (3.8%). Two patients in bi-atrial group received a permanent pacemaker (4.1%). Patients in both groups were free of AF at the end of the procedure. (Bi-atrial group: sinus: 79.2%, pacemaker: 20.8%), (Left atrial group: sinus: 82.5%, pacemaker: 17.5%) ( p > 0.05). During the last follow-up, sinus rhythm was maintained in 79.6% of cases in bi-atrial group, while this rate was 75.6% in left atrial group ( p > 0.05). CONCLUSION: Saline irrigated radiofrequency modified maze procedure was performed safely and efficiently. Both the left and bi-atrial procedures were successful in terms of restoring sinus rhythm. Our current policy is to adopt the bi-atrial approach in patients with a history of atrial flutter and where the right atrium has to be opened. Otherwise the procedure is restricted to the left atrial side. PMID- 14739725 TI - Antiarrhythmic surgery to cure atrial fibrillation--subgroups and postoperative management. AB - Antiarrhythmic surgical procedures to cure atrial fibrillation (AF) are widely used in cardiac surgery. Whereas the Cox maze procedure remains the highly effective gold-standard a variety of different antiarrhythmic procedures aim at reducing the extent and duration of the procedure. Antiarrhythmic procedures are especially effective in patients undergoing mitral valve surgery. In 110 patients with permanent AF undergoing various surgical procedures sinus rhythm was re established in 75%. Subgroup analyses revealed no significant differences in rhythm or survival after antiarrhythmic intraoperative ablation indicating the usefulness and feasibility of this procedure in patients with a wide range of characteristics. Because conversion usually occurs spontaneously within the first 6 months and antiarrhythmic medication does not increase the incidence of conversion it seems reasonable to wait for spontaneous occurrence of sinus rhythm after antiarrhythmic intraoperative ablation. In patients with permanent AF undergoing open heart surgery additional antiarrhythmic procedures have been shown to be safe and effective. PMID- 14739726 TI - How should subgroup analyses affect clinical practice? Insights from the Metoprolol Succinate Controlled-Release/Extended-Release Randomized Intervention Trial in Heart Failure (MERIT-HF). AB - CONTEXT: The Metoprolol CR/XL Randomized Intervention Trial in Chronic Heart Failure (MERIT-HF), the Cardiac Insufficiency Bisoprolol Study II (CIBIS-II), and the Carvedilol Prospective Randomized Cumulative Survival Study (COPERNICUS) have all demonstrated highly significant positive effects on total mortality as well as total mortality plus all-cause hospitalization in patients with heart failure. While none of these trials are large enough to provide definitive results in any particular subgroup, it is of interest for physicians to examine the consistency of results as regards efficacy and safety for various subgroups or risk groups. OBJECTIVE: To summarize results from both predefined as well as post-hoc subgroup analyses performed in the MERIT-HF trial, and to provide guidance as to whether any subgroup is at increased risk, despite an overall strongly positive effect, and to discuss the difficulties and limitations in conducting such subgroup analyses. For some subgroups we performed metaanalyses with data from the CIBIS II and COPERNICUS trials in order to obtain more robust data on mortality in subgroups with a small number of deaths (e.g. for women). SETTING: MERIT-HF was run in 14 countries, and randomized a total of 3,991 patients with symptomatic systolic heart failure (NYHA class II to IV with ejection fraction < or =0.40). Treatment was initiated with a very low dose with careful titration to a maximum target dose of 200 mg metoprolol succinate controlled release/extended release (CR/XL), or highest tolerated dose. MAIN OUTCOME MEASURES: Total mortality (first primary endpoint), total mortality plus all-cause hospitalization (second primary endpoint), and total mortality plus hospitalization for heart failure (first secondary endpoint) analyzed on a time to first event basis. RESULTS: Overall, MERIT-HF demonstrated a 34% reduction in total mortality ( p = 0.00009 nominal) and a 19% reduction for mortality plus all-cause hospitalization ( p = 0.00012). The first secondary endpoint of mortality plus hospitalization for heart failure was reduced by 31% ( p = 0.0000008). The results were remarkably consistent for both primary outcomes and the first secondary outcome across all predefined subgroups as well as nearly all post-hoc subgroups. Metoprolol CR/XL has been very well tolerated, overall as well as in all subgroups analyzed. Overall 87% of the patients reached a dose of 100 mg or more of metoprolol CR/XL once daily, and 64% reached the target dose of 200 mg once daily. CONCLUSION: Our results show that when carefully titrated, metoprolol CR/XL can safely be instituted for the overwhelming majority of outpatients with clinically stable systolic heart failure, with minimal side effects or deterioration. The time has come to overcome the barriers that physicians perceive to beta-blocker treatment, and to provide it to the large number of patients with heart failure in need of this therapy, including also high risk patients like elderly patients, patients with severe heart failure, and patients with diabetes. Because of the increased risk, these are the patients in whom treatment will have the greatest impact as shown by number of lives saved and number of hospitalizations avoided. The target dose should be strived for in all patients who tolerate this dose. We should expect some variation of the treatment effect around the overall estimate as we examine a large number of subgroups due to small sample size in subgroups and due to chance. However, we believe that the best estimate of treatment effect for any particular subgroup should be the overall effect observed in the trial. PMID- 14739727 TI - Quality of life variables in the selection of rate versus rhythm control in patients with atrial fibrillation: observations from the Canadian Trial of Atrial Fibrillation. AB - Many patients with atrial fibrillation develop symptoms attributable to the cardiac arrhythmia itself. These symptoms may be improved either by restoring sinus rhythm or by controlling the rapid and irregular ventricular response that often accompanies this arrhythmia. One of the principal goals of therapy of atrial fibrillation management is improvement of patient symptoms; it is important to quantify these symptoms by some form of quality of life analysis. The Canadian Trial of Atrial Fibrillation (CTAF) was a multi-centre randomized clinical trial of amiodarone compared with either propafenone or sotalol in patients with recent atrial fibrillation. The quality of life (QOL) substudy of CTAF was a prospective, comprehensive assessment of quality of life of patients enrolled in CTAF. Summary measures of physical and mental health on the generic QOL scale (SF-36) improved significantly with treatment from baseline to 3 months (41.9 +/- 9.6 to 43.7 +/- 9.2, p = 0.001 for the physical component and 47.5 +/- 10.4 to 49.0 +/- 9.8, p = 0.023 for the mental component). On an arrhythmia specific scale (SCL), a significant and larger improvement was noted from baseline to 3 months in both arrhythmia symptom frequency and severity (symptom frequency from 20.4 +/- 9.4 to 16.2 +/- 9.5, symptom severity from 16.7 +/- 8.2 to 12.9 +/- 7.6, both p < 0.001). The quality of life improvements were similar in the amiodarone group compared to the sotalol or propafenone groups, both for the SF-36 and the disease-specific symptom checklist (SCL) measures. In contrast, an atrial fibrillation severity scale (AFSS) did show differences between the assigned drug therapies, which were associated with different rates of arrhythmia recurrence in the parent study. By 3 months global well-being was significantly worse for patients who had recurrent atrial fibrillation compared to those who did not (6.9 +/- 1.8 versus 7.4 +/- 1.8, p = 0.04). Similarly, symptom severity at 3 months was 11.8 +/- 7.4 for patients without recurrence, compared to 14.8 +/ 7.4 for those with recurrence ( p = 0.001). Interestingly, none of the usual clinical variables that might be perceived to be associated with quality of life, e.g., male versus female sex, age, NYHA class, beta blocker use, and ejection fraction, had much impact on subjective quality of life measures. Quality of life improves with treatment atrial fibrillation and at least some of these improvements are related to the restoration and maintenance of sinus rhythm. PMID- 14739728 TI - Time to first recurrence as a trial endpoint: time to change? AB - There has been a rapidly increasing interest in developing new therapies for management of atrial fibrillation. The optimal method for evaluating their efficacy is by measuring changes in outcomes such as mortality, quality of life or cost of care; requiring trials of large sample size. In order to reduce the sample size, there is a need to develop appropriate surrogate endpoints. "Is time to first recurrence of symptomatic atrial fibrillation" an appropriate surrogate endpoint for quality of life? Since a surrogate endpoint must capture the net effect of the treatment, it was assumed (a) that frequency of symptomatic episodes captures the net effect on quality of life and (b) "time to first recurrence" is a measure of the frequency of symptomatic episodes. The effect of frequency of symptomatic episodes or their duration and symptom severity on quality of life has not been evaluated. "Time to first symptomatic recurrence" was proposed because data from a few patients demonstrated that symptomatic atrial fibrillation episodes arose independently and randomly and could be represented mathematically by a Poisson distribution. Recent data from a greater number of patients with implantable devices that detect symptomatic and asymptomatic episodes of atrial tachyarrhythmias indicate that these episodes tend to cluster in time and cannot be well represented by a Poisson distribution. Because all atrial tachyarrhythmia episodes do not follow a Poisson distribution, it is unlikely that only symptomatic episodes would follow the same distribution, although this needs to be proved. A non-Poisson distribution requires a larger sample to detect differences in clinical trials. This should be taken into consideration when designing prospective trials. Alternative methods may include measuring frequency of symptomatic episodes as well their severity and duration. In patients with implanted devices, total duration of time spent in atrial tachyarrhythmias or objective measures of rate control should also be evaluated as surrogate endpoints. If surrogate endpoints cannot be developed, changes in clinical outcomes will need to be demonstrated to evaluate therapeutic efficacy. PMID- 14739729 TI - An update on electrical cardioversion of atrial fibrillation. AB - Atrial fibrillation is the most frequently encountered sustained arrhythmia in clinical practice. Electrical cardioversion of atrial fibrillation using damped sine wave shocks has been a mainstay of therapy for nearly 4 decades; its limitation remains a failure rate that approaches 20%. Although several alternatives have been proposed, including delivering 720 J shocks using dual monophasic defibrillators, ibutilide pretreatment and internal cardioversion, each of these approaches has significant limitations, which preclude its routine use. Recent data demonstrate that routine use of biphasic shocks for cardioversion of atrial fibrillation is associated with a marked improvement in cardioversion efficacy and suggest that biphasic shocks may be the preferred method for the transthoracic electrical cardioversion of atrial fibrillation. PMID- 14739730 TI - Biphasic versus monophasic shock waveform for conversion of atrial fibrillation. AB - Cardioversion of atrial fibrillation (AF) using traditional monophasic shock waveform is unsuccessful in up to 20% of cases, and often requires several shocks of up to 360 J. Based on the success with biphasic shock waveform in converting ventricular fibrillation, it was postulated that biphasic shocks would allow cardioversion with lower energy. In a international multicenter, double-blind, randomized trial of 203 patients, damped sine wave monophasic shocks were compared with impedance-compensated truncated exponential biphasic waveform shocks. Patients received up to five shocks: 100 J, 150 J, 200 J, a fourth shock at maximum output for the initial waveform (200 J biphasic, 360 J monophasic) and a final cross-over shock at maximum output of the alternate waveform. For each energy level, the biphasic waveform compared favorably to the monophasic waveform in successful cardioversion (100 J: 60% versus 22%, P < 0.0001; 150 J: 77% versus 44%, p < 0.0001; 200 J: 90% versus 53%, p < 0.0001). Success with 200 J biphasic was equivalent to 360 J monophasic shock (91% versus 85%, p = 0.29). Patients randomized to biphasic waveform required fewer shocks and lower total energy delivered; in addition, this waveform was associated with less dermal injury and no blistering. Biphasic shocks converted AF present for less than 48 hours with 80% efficacy, but conversion of AF present for more than 48 hours and more than 1 year the success rate was only 63 and 20%, respectively. The results of this study is similar to other investigations comparing biphasic and monophasic shock waveforms for conversion of atrial fibrillation. We recommend starting with biphasic energy of 100 J for atrial fibrillation of less than 48 hours duration, but using higher energies (150 J, 200 J or greater) when AF has been present for longer periods. PMID- 14739731 TI - Effect of electrode position on the outcome of cardioversion. AB - Acute termination of atrial fibrillation is the starting point of any therapy aimed at preventing atrial fibrillation and improves cardiac output in patients with hemodynamic compromise due to the arrhythmia. External electrical cardioversion is a simple procedure to terminate persistent atrial fibrillation in the majority of cases. Initially, Lown et al. used an anterior-lateral position of electrodes to apply a cardioversion shock. Recent pathophysiological studies have shown that atrial fibrillation is maintained by functional re-entry circuits anchored in the left atrium. As the left atrium is located posteriorly in the thorax, an anterior-posterior electrode position may be more efficient for external cardioversion of atrial fibrillation than the commonly used anterior lateral electrode position. Several recent studies have confirmed that an anterior-posterior electrode position is superior to an anterior-lateral electrode position for external cardioversion of atrial fibrillation. There are no indications that an anterior-posterior electrode position is less safe than an anterior-lateral electrode position. We therefore suggest that an anterior posterior electrode position should initially be used for external cardioversion of atrial fibrillation. PMID- 14739732 TI - Amiodarone for the restoration of sinus rhythm in patients with atrial fibrillation. AB - Although the use of amiodarone for the treatment of atrial fibrillation has increased, reports of its use for the restoration of sinus rhythm have been conflicting. In a recent prospective, randomized, single-blind, placebo controlled study, we examined the efficacy and safety of amiodarone as initial treatment to restore sinus rhythm in patients with atrial fibrillation of varying duration. We studied 335 patients (169 men, 166 women), aged 27-78 years (mean age 65 +/- 10 years), with symptomatic atrial fibrillation, who presented to the emergency room or to our clinic. Patients randomized to amiodarone ( n = 173) received 300 mg intravenously over 1 hour followed by 20 mg/kg over 24 hours. Oral administration was initiated simultaneously at 600 mg/day in 3 divided doses for one week followed by 400 mg/day, in two doses, for three weeks. Amiodarone was almost twice as likely as placebo to restore sinus rhythm (85% vs. 43%, p < 0.0001). Larger left atrial size and longer duration of atrial fibrillation decreased amiodarone's effectiveness. These factors were also associated with longer duration of treatment before conversion to sinus rhythm. We observed relatively few and only minor complications despite relatively high doses of amiodarone administered to achieve high serum concentrations. In conclusion, our findings indicate that amiodarone is an effective and safe antiarrhythmic drug even when administered in large doses. It may be used for the restoration of sinus rhythm when rapid cardioversion is not needed. PMID- 14739733 TI - Facilitating electrical cardioversion of persistant atrial fibrillation by antiarrhythmic drugs: update on clinical trial results. AB - Results from clinical trials suggest that antiarrhythmic drugs (AD) can facilitate electrical cardioversion (EC) for persistent atrial fibrillation (AF) (duration >48 hours, no spontaneous termination) by suppression of immediate reinitiation of AF following the procedure. Class IC agents may increase the atrial defibrillation threshold (DFT) by significantly reducing the availability of Na+-channel for depolarization. In contrast, class III agents may decrease the atrial DFT by markedly prolonging atrial refractoriness. Among all AD, ibutilide and amoidarone have been shown to be most effective in enhancing the acute outcome of EC. In patients who are over 65 years of age at high risks of stroke (e.g., atherosclerotic cardiovascular disease, diabetes, hypertension, previous thromboembolism, etc.), the rhythm control strategy offers no survival advantage over the rate control strategy and frequently subjects patients to serious adverse effects of AD therapy. It can not be overemphasized that adequate anticoagulation (INR 2.0-3.0) with warfarin is needed regardless of whichever strategy is chosen unless there are contraindications. On the other hand, in patients who are under 65 years of age without structural heart disease or other risk factors of stroke, rhythm control can be the treatment of choice. Specifically, if a patient has failed EC alone or if the patient has characteristics (e.g., duration of AF >6 months, left atrium >50 mm, etc.) that EC could fail, AD may be given before the procedure to facilitate EC. In the subgroup of patients who are symptomatic with hypertrophic cardiomyopathy and severe diastolic dysfunction requiring maintenance of sinus rhythm to have sufficient ventricular function for optimization of cardiac output, an aggressive approach for rhythm control with amiodarone along with adequate anticoagulation with warfarin should be encouraged. PMID- 14739734 TI - Do population studies confirm the benefit of oral anticoagulation in atrial fibrillation demonstrated in clinical trials? AB - Atrial fibrillation (AF) is associated with a significant morbidity and mortality, mainly due to an increased risk of thromboembolic stroke. Several large randomized trials have demonstrated the efficacy of oral anticoagulation to reduce this risk of ischemic events in patients with non-valvular AF. Despite the translation of these results into clear practice guidelines, oral anticoagulation remains underused. This reflects doubts about the real effectiveness of oral anticoagulation in daily practice. This paper gives an overview of the available evidence of the effectiveness of oral anticoagulation in population studies and tries to identify some remaining barriers in the prescription of oral anticoagulation in real life. PMID- 14739735 TI - Barriers to the use of warfarin: potential solutions. PMID- 14739736 TI - Why is warfarin underutilized in patients with atrial fibrillation? PMID- 14739737 TI - Trials of newer approaches to anticoagulation in atrial fibrillation. AB - Atrial fibrillation comprises a large and growing epidemic in the aging population. Stroke is the most feared complication of atrial fibrillation, and the risk of stroke increases markedly with age. Multiple clinical trials have proven that warfarin anticoagulation is effective in reducing this risk. However, the complex pharmacokinetics and narrow therapeutic window of warfarin make its use in clinical practice challenging. Novel approaches to anticoagulation, including more potent antiplatelet agents and direct thrombin inhibitors, are currently undergoing clinical trials. In addition, nonpharmacological approaches to stroke prevention in atrial fibrillation are also in development. These newer approaches may revolutionize the treatment of this common disorder. PMID- 14739739 TI - Randomized trials of rate vs. rhythm control for atrial fibrillation. AB - Recent randomized trials have not demonstrated mortality or stroke risk reduction benefits from a rhythm-control compared to rate-control strategy in the treatment of atrial fibrillation. These studies reinforce the need for continued anticoagulation in both strategies for patients with atrial fibrillation and risk factors for stroke. Although rate control can be rationalized as a first line approach, rhythm control strategies may be justified for patients who are younger, who remain symptomatic or functionally impaired, or who have a first episode of atrial fibrillation. PMID- 14739738 TI - Alternatives to warfarin for thromboembolism prophylaxis in nonrheumatic atrial fibrillation. AB - Decision-making regarding thromboembolism prophylaxis in atrial fibrillation remains a major clinical challenge. While evidence of the beneficial effect of anticoagulation for patients participating in clinical trials is well established, only half of eligible individuals in the general population are currently treated with warfarin. Using an evidence-based approach, this review covers major therapeutic approaches in practice today and many of those expected to be released in the near future. Pharmacologic agents evaluated include warfarin, aspirin, other antiplatelets agents, direct thrombin inhibitors and antiarrhythmic drugs. Nonpharmacologic treatments reviewed include surgical and catheter ablation, pacing, left atrial appendage ligation and occlusion methods, and atrial defibrillators. PMID- 14739740 TI - Clinical trials of pacing for maintenance of sinus rhythm. AB - Several prospective randomized clinical trials have demonstrated that atrial based pacing prevents the development of paroxysmal and persistent atrial fibrillation (AF) in patients with symptomatic bradycardia as the indication for a pacemaker. The greatest benefit appears to be in patients with sinus node dysfunction as the primary indication for pacing. Studies of site specific atrial pacing have not shown a consistent benefit for prevention of AF. A number of prospective studies have been undertaken to assess the role of selective atrial pacing algorithms designed for prevention of AF. These studies suggest that these AF pace prevention algorithms have modest to minimal incremental benefit compared to atrial based pacing alone for the prevention of AF. Atrial antitachycardia pacing (ATP) algorithms have been incorporated into some pacemakers and defibrillators. Overall, atrial ATP therapy has not been shown to substantially reduce the burden of AF in one randomized trial. However, there are subsets of patients e.g. those with atrial flutter as well as AF who are likely to benefit from this therapy. PMID- 14739741 TI - Trials of pacing to control ventricular rate during atrial fibrillation. AB - Pharmacologic therapy to achieve rate control in patients with atrial fibrillation is often difficult and inadequate. For this reason, ventricular pacing strategies have been developed as an alternative to drug therapy to alleviate symptoms due to rapid and irregular ventricular rates. Ventricular pacing in combination with AV junctional ablation provides palliative improvement in a wide range of clinical outcomes. Because of the irreversible complete AV block associated with this procedure, strategies to control the ventricular response to atrial fibrillation by ventricular pacing alone have been investigated. These strategies are primarily directed at regularizing the ventricular response by pacing at or near the mean intrinsically conducted ventricular rate. These specialized ventricular pacing algorithms provide striking ventricular regularity at rest but may be less effective during activity. No study has yet demonstrated clinically significant improvements in clinical outcomes with these algorithms. The clinical benefits of rate regularization alone without the strict rate control provided by AV junctional ablation are likely to be very limited. Other device based approaches to control ventricular rate in atrial fibrillation include transvenous vagal stimulation. This strategy is in early stages of development but may be promising. PMID- 14739742 TI - Trials of new antiarrhythmic drugs for maintenance of sinus rhythm in patients with atrial fibrillation. AB - Despite the recent results of the AFFIRM trail showing that either strategy of rate control or rhythm control is acceptable, there is general agreement that therapy must be individualized. Antiarrhythmic drugs while being effective in maintaining sinus rhythm are not without side effects. Thus, there continues to be a search for safer and better tolerated agents. Azimilide is a novel class 3 agent that blocks both components of the delayed rectifier current, while dofetilide blocks only the slow component. Dronedarone is somewhat similar to aminodarone, but perhaps with the side effects. Tedisamil is a complex class 3 agent, blocking multiple channels. Piboserod is a selective 5-HT receptor antagonist that is being investigated in clinical trials. PMID- 14739743 TI - Advances in the treatment of atrial fibrillation: the future is now. PMID- 14739744 TI - Variability of AV nodal potentials recorded, in vivo: direct demonstration of dual AV nodal physiology. AB - OBJECTIVES OF STUDY: We developed a method to record extracellular A-V nodal potentials in the beating dog heart, in vivo. METHODS: In eleven Na-pentobarbital anesthetized, open-chest dogs, an octapolar electrode catheter (2 mm rings, 2 mm spacing) was inserted through a purse-string suture in the coronary sinus (CS) distal to the ostium and positioned electrographically so that the tip electrode recorded a His bundle (Hb) potential. RESULTS: Stable recordings of A-V nodal potentials (amplitude, 178 +/- 94 microV; duration 78 +/- 26 msec) were consistently made during sinus rhythm from the second and/or third bipolar pairs of electrodes. Programmed atrial stimulation and vagal stimulation resulted in loss of amplitude and increased duration of the A-V nodal potentials associated with A-H prolongation. In another series of experiments, crushing the sinus node in 6 dogs resulted in AV nodal rhythms with AV nodal potentials of varying amplitudes (132 to 840 microV) and durations (range 25 to 71 msec) as the earliest activation which preceded the Hb, atrial and ventricular deflections. One dog, showing dual AV nodal physiology as documented from the AV nodal function curve, had two distinctly different AV nodal potentials. The low-level, longer duration potentials were associated with longer (slow pathway) A-H intervals; whereas the shorter higher amplitude potentials (fast pathway) showed shorter A-H intervals, each occurring at a critical paced cycle length. CONCLUSION: We conclude that consistent extracellular AV nodal electrograms can be recorded in vivo although the configuration of these potentials varies depending on heart rate, autonomic stimulation and different arrhythmic conditions such as AV nodal escape rhythms and dual AV nodal physiology. PMID- 14739745 TI - Relative utility of magnetic resonance imaging and right ventricular angiography to diagnose arrhythmogenic right ventricular cardiomyopathy. AB - INTRODUCTION: Arrhythmogenic right ventricular cardiomyopathy (ARVC) is characterized by fibrofatty replacement of the RV myocardium. Two imaging techniques used to assess patients suspected of having ARVC are magnetic resonance imaging (MRI) and right ventricular angiography (RVA). Traditionally, RVA has played a central role in the diagnosis of ARVC, but the non-invasive nature of MRI and its unique ability to detect fatty tissue infiltration has increased its popularity as a diagnostic tool. The objective of this study was to assess the relative diagnostic accuracy of MRI and RVA for ARVC. METHODS AND RESULTS: Seventeen patients (9 men, 8 women; ages 42 +/- 17 [range 16-78] years) with documented ventricular arrhythmias were investigated for ARVC. A positive diagnosis of ARVC was based on criteria set forth by the ISFC Working Group on Cardiomyopathies and Dysplasia. ECG-gated spin-echo and gradient-echo MR images in multiple planes and RAO/LAO RV angiograms were compared for diagnostic concordance. Based on working group criteria, 7 patients were diagnosed with ARVC. In ten patients, MRI suggested ARVC. The remaining 7 patients had no MRI findings suggestive of the disease. Four patients with MRI findings of ARVC were incorrectly diagnosed based on Task Force criteria. Conversely, 1 patient with a normal MRI met Task Force criteria for the diagnosis of ARVC. Based on RV angiograms, 7 patients had findings suggestive of ARVC. The 10 patients without AVRD (based on RVA) also did not meet the necessary criteria for diagnosis of ARVC using Task Force standards. RVA was 100% specific and 100% sensitive compared to MRI that was only 86% sensitive and 60% specific. MRI proved to be most reliable when the images demonstrated gross, lipomatous infiltration, evidenced by a large area of hyperintensity. When the results of MRI and RVA were congruent, the diagnosis was always accurate. CONCLUSION: RVA is more sensitive and specific to diagnose ARVC diagnosis than MRI, at least until MRI protocols are better developed. MRI results are most robust when indicators of ARVC are grossly apparent. False-positive diagnosis by MRI was primarily related to perceived motion abnormalities that were not seen by RVA. One of its greatest potential assets (fat detection) did not enhance diagnostic specificity. PMID- 14739746 TI - Magnetic resonance imaging in the diagnosis of arrhythmogenic right ventricular cardiomyopathy: the gold standard or just another imaging modality? PMID- 14739747 TI - Feasibility and accuracy of pre-procedure imaging of the proximal cephalic vein by duplex ultrasonography in pacemaker and defibrillator implantation. AB - BACKGROUND: Failure of the cephalic venous approach in pacemaker and defibrillator implantation is always due to the small size and difficulty in isolation of the cephalic vein. We propose that pre-procedure imaging of the proximal cephalic vein is valuable to achieve successful access of cephalic vein. However, the feasibility and accuracy of duplex ultrasonographic imaging of the proximal cephalic vein are unknown. METHODS: The study enrolled 30 consecutive patients who underwent new implantation of permanent pacemakers or defibrillators at our institute. An ultrasound probe scanned along the plane 2 cm beneath the inferior margin of the clavicle to locate the cephalic vein before device implantation. If the vein was well visualized, the venous diameter and the vertical depth were measured. The corresponding surface location of the vein on the chest wall was also identified and recorded by duplex ultrasonography. The echo-derived vertical depths and vascular findings were compared with those measured during surgery. RESULTS: All proximal cephalic veins were well visualized in the infraclavicular region by duplex ultrasonography. They were compressible, patent in color Doppler ultrasound imaging, and displayed phasic change of Doppler signal during respiration, indicating patency in all study veins. The average diameter of the target cephalic vein was 7.7 +/- 1.6 mm (range, 5.0-11.1 mm). The echo-derived vertical depth of the proximal cephalic veins was highly correlated with the depth measured during surgery (28.4 +/- 5.5 vs. 28.4 +/- 5.6 mm, r = 0.93, P < 0.0001). All target cephalic veins were isolated after exploration via the estimated surface location of the chest wall by pre-procedure duplex ultrasonography. Seven (23%) of the studied patients did not have their cephalic vein cannulated successfully. CONCLUSION: The target proximal cephalic vein in pacemaker and defibrillator implantation can be precisely imaged and localized by duplex ultrasonography. Although further studies are needed, our findings pave a way to further study and clarify the implantation problems of cephalic vein approach. PMID- 14739748 TI - Triple-site pacing in patients with biventricular device-incidence of the phenomenon and cardiac resynchronization benefit. AB - BACKGROUND: In patients with biventricular pacing (BIV), triple-site pacing (TSP), i.e. standard biventricular cathodal pacing of the right and the left ventricle plus additional anodal capture of the right ventricle, is sometimes present. AIMS: To evaluate the incidence of TSP phenomenon, to examine TSP related QRS changes, and to assess the effect of TSP on intraventricular resynchronization by means of tissue Doppler imaging (TDI). METHODS AND RESULTS: 23 patients with a first generation biventricular device (Medtronic 8040) and 16 patients with a new generation device (Medtronic 8042) were evaluated to look for the presence of TSP. TSP was found in 6 patients (26%) with the Medtronic 8040 (group I) and in 13 patients (81%) with the Medtronic 8042 device (group II). QRS duration decreased by 10 to 20 ms and QRS amplitude of leads I and aVL increased in almost all patients in group I during TSP modality. In group II, QRS morphology, duration and amplitude did not change as obviously. TDI analysis of the left ventricular (LV) basal segments showed significant shortening of the systole, together with a corresponding prolongation of the diastole, at the inferior wall of the LV, during TSP compared to standard BIV in all patients ( p < 0.01). Other LV segments did not show any change. Qualitative TDI electro mechanical activation pattern of all LV segments improved in 22%, while it remained unchanged in 72%. CONCLUSIONS: TSP phenomenon can be identified in approximately a quarter of patients with the first-generation biventricular devices on the basis of the QRS morphology changes. In the second-generation biventricular pacemakers it can be demonstrated in the vast majority of patients. TSP may increase the effectiveness of cardiac resynchronization therapy by counteracting the regional activation delay located at the inferior wall of the LV. PMID- 14739749 TI - Loss of left bundle branch block following biventricular pacing therapy for heart failure: evidence for electrical remodeling? AB - Cardiac resynchronization therapy has been proven to improve symptoms and indices of left ventricular function in patients with heart failure and intraventricular conduction delays. We present a case of a patient with New York Association Class III heart failure and left bundle branch block, who received a biventricular pacemaker in order to achieve cardiac resynchronization. Her symptoms improved markedly, as did left ventricular ejection fraction and dimensions. In addition, her intrinsic QRS duration normalized. This may represent a salutory effect of biventricular pacing on electrical remodeling. PMID- 14739750 TI - Conduction patterns in the cardiac veins: electrophysiologic characteristics of the connections between left atrial and coronary sinus musculature. AB - INTRODUCTION: Fractionated electrograms and double potentials have been well described within the coronary sinus (CS) in humans. The pattern of circumferential activation in the CS has not been investigated. Furthermore, no data exist on conduction characteristics within the great cardiac vein (GCV) or the middle cardiac vein (MCV). METHODS AND RESULTS: Twenty patients underwent catheter mapping of the CS, the MCV, and the GCV. Anatomical areas were verified by cannulation of the left superior pulmonary vein. The pattern of circumferential muscle activation within the proximal CS was also studied with a circular mapping catheter (Lasso 12 mm). At conventional mapping during sinus rhythm and high right atrial pacing, discrete double potentials or fractionated electrograms were recorded during left, right atrial and CS pacing at the CS ostium, mid-CS, and distal CS-ligament of Marshall area, in 2 (10%), 1 (5%), and 9 (45%) patients, respectively, whereas no patient displayed such signals in the MCV or GCV ( p < 0.001). Proximal CS mapping with the Lasso was accomplished in 10 patients, 7 of whom had no evidence of multicomponent potentials in the CS at conventional mapping. Specific CS potentials dissociated from the atrial electrograms were recorded in all patiens with the use of circumferential mapping. The perimetric distribution of electrograms within the CS suggested an oblique course of conduction across the CS musculature. CONCLUSION: Potentials representing activation of the CS musculature, with an oblique course of conduction across the CS, can be recorded in human CS but not in the GCV or MCV. This is compatible with anatomical observations of sinus venosus musculature covering the CS but not other cardiac veins, and supports the rationale for the role of CS musculature in the generation of atrial arrhythmias. PMID- 14739751 TI - Effect of left atrial-coronary sinus musculature connections on the coronary sinus activation pattern via retrograde conduction in patients with WPW syndrome. AB - BACKGROUND: Double potential (DP) activation patterns observed in coronary sinus (CS) electrograms recorded during left lateral atrial pacing, were explained by an initial low-frequency left atrial (LA) activation potential and secondary high frequency CS musculature activation potential in canine hearts. Moreover, the connections between the LA and CS musculature vary greatly in size and location in the human heart. The purpose of this study was to investigate the relationship between the CS activation pattern during retrograde conduction via an accessory pathway (AP) and the location of left-sided APs. METHODS AND RESULTS: Fifty-one patients (31 males, mean age 48.6 years) who underwent radiofrequency catheter ablation of left-sided APs were divided into two groups according to the successful ablation site. The CS electrograms during retrograde AP conduction were classified into 3 types; single, fractionated, and DP activation patterns. A DP pattern was identified in 10 of 12 patients (83.3%) with posteroseptal to posterolateral APs, and in particular, 9 had a divergent sequence. Twenty-six of 39 patients (66.7%) with lateral to anterolateral APs, demonstrated a single pattern. The number of radiofrequency applications was significantly higher in patients with a DP pattern than in those with a single pattern (3.4 +/- 3.3 vs. 7.8 +/- 6.8, p < 0.01). CONCLUSION: Misleading information obtained when mapping for optimal ablation sites might result from DP patterns with a divergent sequence produced by discrete muscular connections between the LA and CS musculature. Ablation around left posterior APs may require meticulous observation of the CS activation patterns. PMID- 14739752 TI - Relative timing of isolated potentials during postinfarction ventricular tachycardia and sinus rhythm. AB - BACKGROUND: In postinfarction patients, isolated potentials separated by an isoelectric segment from the ventricular electrogram indicate areas of block. Isolated potentials can be recorded during both sinus rhythm and ventricular tachycardia (VT). In an attempt to differentiate bystander pathways from critical sites within a reentry circuit, we compared the relative timing of isolated potentials during VT and sinus rhythm. METHODS: In 19 patients (mean age 68 +/- 6 years) with postinfarction VT who were referred for VT ablation, mapping was performed in the presence and absence of VT. Forty-three sites at which there was concealed entrainment during 35 VT's (mean cycle length 469 +/- 74 ms) displayed an isolated potential separated from the main portion of the ventricular electrogram by an isoelectric segment of >/=30 msec in the presence and absence of VT. The interval between the ventricular electrogram and the isolated potential was measured during VT and baseline rhythm, and the absolute difference (DeltaIPI) was calculated. The DeltaIPI was significantly greater at effective ablation sites (119 +/- 69 ms) than at ineffective ablation sites (30 +/- 28 ms, p < 0.001). The positive predictive value of a DeltaIPI > 85 ms for an effective ablation site was 100%. CONCLUSION: At sites of concealed entrainment, an absolute difference >85 ms between the isolated potential intervals during sinus rhythm and VT is highly specific for a critical area of the VT reentry circuit in post-infarction patients. PMID- 14739753 TI - Persistent tachycardia with a 2:1 exit block within an isolated pulmonary vein. AB - We describe a patient with drug-resistant, paroxysmal atrial fibrillation who underwent segmental pulmonary vein (PV) isolation. After complete isolation of the right superior PV, a persistent regular tachycardia was recorded within the vein. A tachycardia focus with a cycle length of 114 ms was found 2.5 cm away from the ostium of the PV. The cycle lengths of the PV tachycardia near the ostium and near the focus were 400 ms and 200 ms, respectively, which indicates the presence of a 2:1 exit block within the vein. This PV tachycardia was completely eliminated with the application of radiofrequency energy at the focus. PMID- 14739754 TI - The circadian variation of atrial defibrillation thresholds. AB - BACKGROUND: A circadian variation exists for ventricular defibrillation thresholds (DFTs) with a morning peak and a corresponding decrease in therapy success rates from implantable cardioverter defibrillators. Such a variation in atrial DFTs may have implications for the timing of internal cardioversion of atrial arrhythmias. The aim of this study was therefore to determine the circadian variation of atrial DFTs in patents with recurrent atrial fibrillation (AF). METHODS AND RESULTS: Data were collected as part of the worldwide Jewel AF only study. Patients had recurrent persistent AF and no history of ventricular arrhythmias. The atrial DFT was assessed at device implantation using a step-up protocol and was recorded for 100 patients (age 63.0 +/- 11.7, 74% male, ejection fraction 49.6 +/- 17.8%, left atrial diameter 46 +/- 9 mm). The mean atrial DFT was 6.3 +/- 4.3 J. For the most commonly tested lead configuration (right atrium to coronary sinus in 56 patients), the atrial DFT for patients implanted in the morning (3.3 +/- 1.5 J) was significantly lower than for both the DFT measured in the afternoon (5.8 +/- 3.4 J, p < 0.01) and the DFT measured in the evening (7.4 +/- 5.9 J, p < 0.01). CONCLUSION: There may be a significant variation in measured atrial DFT for the right atrium to coronary sinus configuration, with a nadir in the morning. This is the converse to measurements of ventricular DFTs suggesting different regulatory electrophysiological mechanisms. Further investigation of this possible variation is warranted. PMID- 14739755 TI - Dual defibrillator improves quality of life and decreases hospitalizations in patients with drug refractory atrial fibrillation. AB - AIM OF THE STUDY: to evaluate the impact of dual defibrillator implantation on quality of life and resource utilization in patients with drug refractory atrial fibrillation (AF) without prior ventricular arrhythmias. METHODS: Forty patients (28 M, mean age 64 +/- 10) received a dual defibrillator Medtronic 7250. AF was persistent in 60% and paroxysmal in 40%. RESULTS: The follow-up lasted 15 +/- 4 months (range 12-30). Eighty-five percent of patients had atrial tachyarrhythmia recurrences. Among 1366 treated episodes, overall success rate was 60.1% for antitachy pacing and 88.2% for atrial shock. Within one year after implant, arrhythmia related hospitalization number decreased from 1.5 +/- 2.0 to 0.4 +/- 0.8 ( p < 0.01) and 77% of patients were free from hospitalization. As regard to quality of life, Symptom Checklist/Frequency and Severity Scale improved after implant for all items and SF-36 questionnaire showed significant improvements in physical activities because of health problems and social activities. The patients assigned to early delivery of atrial shock after AF onset, when compared with the patients who did not accept atrial shock, showed a significant reduction of AF burden, a higher reduction of hospitalization number and a greater improvement of quality of life. CONCLUSION: Dual defibrillator improved quality of life and decreased resource utilization in patients with drug refractory AF. Early delivering of atrial shock seems to be the most effective option. PMID- 14739756 TI - Demetrio Sodi-Pallares (1913-2003). PMID- 14739757 TI - Swedish medical students' views of the changing professional role of medical doctors and the organisation of health care. AB - Medical students will influence future health care considerably. Their professional orientation while at medical school will be related to their future professional development. Therefore, it is important to study this group's view of the role of medical doctors, especially because Swedish health care is currently undergoing major changes and financial cut backs. Here, the theoretical framework was contemporary theories of competence development, which has shown that people's understanding of their work influences their actions. The aim of this study was to describe medical students' views of their future professional role in health care. In total, 57 fourth-year medical students at a Swedish university were asked to write a short essay about how they conceptualised their professional role in future health care. Fifty-three students (93%) replied. The essays were analysed qualitatively in three steps and four themes were subsequently identified: the professional role in change, organisation of health care, working conditions and the possibilities of having a balanced life. Some factors mentioned that would strongly influence the professional role were being team leader, increased specialisation, supporting the patient and computer science and technology. The students expressed ambiguous feelings about power and leadership. The results indicate that the students share a rather dark view of both the medical profession and health care, which seems to be related to stress and financial cut backs. Mentoring, time for reflection and changes in the curricula might be needed. PMID- 14739758 TI - A controlled trial of active versus passive learning strategies in a large group setting. AB - OBJECTIVE: To compare the effects of active and didactic teaching strategies on learning- and process-oriented outcomes. DESIGN: Controlled trial. SETTING: After hours residents' teaching session. PARTICIPANTS: Family and Community Medicine, Internal Medicine, and Pediatrics residents at two academic medical institutions. INTERVENTIONS: We randomly assigned residents to two groups. One group received a didactic lecture on effective use of diagnostic tests; during this session, the teacher spent a full hour delivering content. The other group received the same content in a session structured to foster resident-to-resident interactions. In the latter session, the teacher spent only 30 minutes directly delivering content to residents. MEASURES AND MAIN RESULTS: We measured residents' knowledge about and attitudes toward the session content before, immediately after, and one month after each session. We measured residents' perceptions of engagement and session value immediately after each session. We employed blinded observers who used a structured instrument to observe residents' activities during each session. Both teaching methods led to improvements in residents' scores on both knowledge and attitude assessments. The amount of improvement was not statistically different between groups. Residents in the active learning session perceived themselves, and were observed to be, more engaged with the session content and each other than residents in the didactic session. Residents in the didactic session perceived greater educational value from the session compared to residents in the active session. CONCLUSIONS: We reduced the amount of time spent in teacher driven content delivery by 50 percent and covered the same amount of content with no detrimental effects on knowledge acquisition or attitude enhancement. Teaching strategies that foster learner-to-learner interactions will lead to more active engagement among learners, however, these learners may value the session less. Further research is needed to explore learner perceptions of the teaching process and other outcomes of active learning in medical education. PMID- 14739759 TI - Detecting score drift in a high-stakes performance-based assessment. AB - Although studies have been conducted to examine the effects of a variety of factors on the comparability of scores obtained from standardized patient examinations (SPE), little research has been conducted to specifically investigate the challenge of detecting drift in case difficulty estimates over time, particularly for large-scale, performance-based, assessments. The purpose of the current study was to investigate the use of a procedure to detect drift in the difficulty estimates for a large-scale, high stakes SPE. The results of this investigation suggest that, for particular performance tasks, there was some variation in mean scores over time. These findings indicate that, although it is feasible to create a bank of case-SP means and link scores back to these fixed estimates, special attention must be paid to the standardization of exam materials over time. This is essential to ensure comparability of scores and pass fail decisions for candidates who are assessed on multiple test forms throughout the year. PMID- 14739760 TI - Using a longitudinal database to assess the validity of preceptors' ratings of clerkship performance. AB - PURPOSE: To examine the validity of using scores from a clinical evaluation form as an assessment of clinical competence. METHOD: Investigators collected a longitudinal clinical skills assessment database that included scores reflecting performance on standardized patient interactions, case-based learning performance, scores on multiple-choice clinical examinations, and preceptors' ratings of students during their clerkships. Pearson correlation coefficients and coefficients corrected for attenuation were calculated between the mean preceptor rating CEF score and the other measures collected during years one and two. RESULTS: Estimates from an earlier across-clerkship generalizability study of the CEF demonstrated that a mean rating computed across 28 forms and four clerkships (the average measurement frequencies observed in this study), yielded an estimated G-coefficient of 0.62 (Kreiter and Ferguson, 2001). Reliabilities for the other measures ranged from 0.23 to 0.56. Nine of the twelve clinical skill measures correlated with the CEF. For those significantly correlated measures where reliabilities could be calculated, correlations corrected for attenuation ranged from 0.46 to 0.58. CONCLUSION: This study indicates that the skills measured by the CEF are related to other clinical performance measures and, conversely, that pre-clinical measures of skills that are believed to be important during clinical years are in fact predictors of preceptors' ratings of clinical performance later in medical school. In addition, the magnitude of the disattenuated coefficients suggests that ratings on the CEF are dependent on important aspects such as clinical knowledge. This study suggests that when averaged over a large number of observations, mean CEF scores demonstrate validity coefficients large enough to support their use as part of an evaluation of students' clinical performance. PMID- 14739761 TI - Assessing the written communication skills of medical school graduates. AB - The ECFMG Clinical Skills Assessment (CSA) was developed to evaluate whether graduates of international medical schools (IMGs) are ready to enter graduate training programs in the United States. The patient note (PN) exercise, conducted after a 15-minute interview with a standardized patient (SP), is specifically used to assess a candidate's ability to summarize and synthesize the data collected. On a yearly basis, approximately 75,000 patient notes are reviewed and scored by physician raters. Recent changes to the PN scoring rubric, combined with enhancements to quality assurance procedures, mandate that additional evidence be provided to support the intended use of PN scores. The purpose of this study was to further investigate the psychometric adequacy of PN scores. Generalizability analyses suggest that while variability in PN ratings can be attributed to the choice of rater, candidate scores are reproducible over the 10 encounter CSA. The relationship of PN scores with other related ability measures and select candidate characteristics provides additional evidence to support the validity of the written exercise. PMID- 14739762 TI - Self-directed learning and student pragmatism. AB - OBJECTIVE: To explore the experience and practice of students entering a problem based (PBL) medical undergraduate course and to identify contributory social, curricular and contextual factors. DESIGN: A multiple case study exploiting the natural experimental features of the setting. Qualitative data collection methods, including participant observation, interviews and focus groups were supplemented by a survey. SETTING: The medical school at the University of Liverpool between 1996 and 1998, which launched an integrated PBL curriculum in line with recommendations of the General Medical Council of the UK. PARTICIPANTS: Two successive cohorts of first year medical students entering the PBL course. RESULTS: The first case study findings demonstrated student insecurity and dependence on the faculty resources rather than conforming to PBL principles. Uncertainty gradually diminished but resource dependence persisted, endorsed by peer interaction. Survey results confirmed the qualitative findings. The hypotheses raised for the second case study predicted replications of resource dependent learning, social influences and initial insecurity. Only the first two were upheld. Second year students' advice on textbooks undercut initial uncertainty but insecurity and social comparison reappeared following the changeover of PBL groups and tutors in the second semester. Survey results were replicated. CONCLUSION: Student learning was socially agreed amongst the peer group and directed by faculty given resources. It was not self-directed. Claims made for PBL type curricula may not be justified and GMC recommendations may remain unfulfilled without rigorous attention to educational principles. PMID- 14739763 TI - Back to the future. AB - I attended all 42 hours of classes in the Cardiovascular System Course in order to help the course coordinator set the examination. What makes this unusual is that I am not a physician. As a PhD educator supporting faculty and the educational programs at our medical school I became a learner as well as a co course co-coordinator. Attending all the classes has led me to discover what might become the future of faculty development and course renewal. I now know who teaches what and how so I can approach them to talk about any changes they may want to make in their teaching. I have also learned what the specific content of the course is which will allow me to connect with coordinators of concurrent and of supporting courses from earlier years in the hope of achieving some higher degree of integration. Taking the course with medical students has been one of the most interesting and professionally valuable tasks that I have ever undertaken, one that will be repeated. PMID- 14739764 TI - MMP induction and inhibition in myocardial infarction. AB - Short-term survival following a myocardial infarction (MI) has greatly improved, due in part to therapeutic interventions that restore blood flow and limit infarct size. The increased incidence of infarct-stimulated left ventricular (LV) remodeling that advances to congestive heart failure (CHF), however, is a significant long-term complication and a leading cause of mortality. Changes to ECM structure and function are primary components of LV remodeling and are precipitated by the early increase in infarct area collagen levels that replace necrotic myocytes and form a scar. ECM turnover is coordinated through the synthesis and degradation of ECM and non-ECM components, particularly the matrix metalloproteinases (MMP), a family of proteolytic enzymes that cleave ECM. MMPs have multiple roles in remodeling events that lead to LV dilation. The inhibition or targeted deletion of specific MMPs attenuates LV remodeling events post-MI. MMP inhibitors have been used in animal models to delineate LV remodeling mechanisms and to evaluate the pharmacologic potential of targeting the ECM to modify LV remodeling post-MI. This review summarizes the current knowledge and limitations of MMP inhibition in the post-MI myocardium. PMID- 14739766 TI - The dynamic interaction between matrix metalloproteinase activity and adverse myocardial remodeling. AB - The process of cardiac remodeling in response to cardiac injury and/or persistent elevations in wall stress generally relates to the progressive changes that occur in ventricular chamber dimensions and the various components of the myocardium, in particular the cardiomyocytes and the extracellular matrix. Volume overload, pressure overload or myocardial injury produces a sustained abnormal elevation in myocardial wall stress which initiates cardiac remodeling that frequently results in ventricular decompensation and heart failure. Regardless of the inciting cause, there appear to be three distinct phases to this process. In the initial phase, fibrillar collagen is partially degraded secondary to increased matrix metalloproteinase (MMP) activity. Following this, there is a chronic compensatory phase during which MMP activity and collagen concentration return to normal while cardiomyocyte size continues to progressively increase. The final phase is attained once the compensatory hypertrophic mechanisms are exhausted and is characterized by elevated MMP activity, marked ventricular dilatation and prominent fibrosis. Details of this progressive, dynamic remodeling process and its effect on ventricular function during chronic volume overload, chronic pressure overload and following myocardial infarction will be the focus of this article. PMID- 14739765 TI - Myocardial remodeling in viral heart disease: possible interactions between inflammatory mediators and MMP-TIMP system. AB - Matrix metalloproteinases (MMP), a family of proteases, are involved in the degradation of extracellular matrix proteins and hence in the determination of interstitial architecture. In the heart, MMPs have been found to play a significant role in the development of myocardial remodeling and congestive heart failure. Tissue inhibitors of matrix metalloproteinases (TIMPs) represent a family of proteins which are known to regulate the expression and activity of MMPs. TIMPs are endogenous physiological inhibitors of MMPs and their concomitant downregulation in heart failure suggests the existence of a critical balance between MMPs and TIMPs in the normal maintenance of myocardial interstitial homeostasis. In addition, cytokines regulate expression of both MMPs and TIMPs besides eliciting a direct effect on myocardial cell function. Therefore, myocardial inflammation may also contribute to the development of cardiac remodeling along with other stimuli like mechanical stress and humoral factors. Viral myocarditis, a predisposing factor for dilated cardiomyopathy, is a condition in which extent of intramyocardial inflammation is thought to determine the progression of disease. Inflammatory events in the heart following viral infection are speculated to be responsible for the transition of myocarditis to dilated cardiomyopathy. In viral myocarditis and other inflammatory heart diseases, the inflammatory cells and their battery of cytokines may also alter the myocardial MMP-TIMP system and eventually lead to dilation of the heart and ventricular dysfunction. The objective of this review is to present an overall picture of the inflammatory phase in viral myocarditis and discuss the possible interactions between inflammation and myocardial MMP profiles which may lead to the evolution of dilated cardiomyopathy. PMID- 14739767 TI - Regulation of matrix metalloproteinases by cytokines and reactive oxygen/nitrogen species in the myocardium. AB - Dysregulation of the myocardial extracellular matrix contributes to abnormal cardiac muscle function. Changes in the balance between matrix deposition and matrix degradation by matrix metalloproteinases (MMPs) can lead to cardiac fibrosis and dilation. In this review, we discuss the regulation of MMPs, their endogenous inhibitors (TIMPs) and collagen synthesis by inflammatory cytokines and reactive oxygen/nitrogen species (ROS/RNS). Inflammatory cytokines, such as interleukin-1beta and tumor necrosis factor-alpha, and ROS activate mitogen activated protein kinases and stress-responsive protein kinases in cardiac cells. In non-cardiac tissues, inflammatory cytokine activation of these kinases is redox sensitive, suggesting ROS may also be involved in cytokine signaling in the heart. Subsequent activation of transcription factors including AP-1, Ets, and nuclear factor kappa-B leads to increased transcription of MMPs. ROS also directly activate MMPs post-translationally. In addition, inflammatory cytokines and ROS lead to decreased TIMP levels and collagen synthesis. Work in animal models suggests that inhibition of inflammatory cytokine or ROS signaling leads to less myocardial remodeling. Further study of the signaling of regulation of the cardiac extracellular matrix may lead to new approaches for the treatment of myocardial remodeling and failure. PMID- 14739770 TI - [Optimal topical treatments for atopic dermatitis]. PMID- 14739768 TI - Matrix metalloproteinases: pathways of induction by bioactive molecules. AB - Regulation of the extracellular matrix (ECM) is an important therapeutic target that can potentially attenuate the adverse ventricular remodeling seen in the progression of heart failure. Matrix metalloproteinases (MMPs) degrade numerous ECM proteins. Importantly, the activation of MMPs and their endogenous inhibitors (TIMPs) are associated with ventricular remodeling. Bioactive-molecules (vasoactive peptides) become activated in proportion to the magnitude of heart failure and have been demonstrated to affect directly collagen degradation as well as collagen synthesis in the myocardium. Pro-fibrotic factors such as norepinephrine, angiotensin II, and endothelin-1 stimulate fibrosis by modulating collagen synthesis and MMP/TIMP activity. Antagonism of these bioactive-molecules has produced improved hemodynamic performance concomitant with modulation of MMP/TIMP activity and in association with reverse remodeling. The natriuretic peptides and nitric oxide, both of which function via the second messenger cGMP, demonstrate anti-fibrotic actions by inhibiting collagen synthesis and by stimulating MMP activity. Furthermore, bioactive-molecules along with certain cytokines are reported to amplify MMP activity, suggesting that different signaling systems work together to modulate ECM turnover. Taken together, the evidence supports an important functional role for bioactive-molecules in the regulation of ECM turnover and suggests that pharmacological intervention at the level of such bioactive molecules may provide potential therapeutic strategies for attenuation of the adverse ventricular remodeling associated with the progression of heart failure. PMID- 14739771 TI - [Various problems of elimination diet and future to oral tolerance in childhood food allergy]. PMID- 14739769 TI - Matrix metalloproteinase inhibitor development and the remodeling of drug discovery. AB - Collagen turnover is a slow process on a biologic timescale with a t$??frac12$ of 20-27 days that is mediated primarily by the matrix metalloproteinases (MMPs). Low collagen metabolism is not due to an intrinsically low Km of MMPs, but rather due to a highly regulated system of activity. Despite the stability of collagen and MMPs, the articles in this special addition illustrate the importance of this enzyme family in the disease process leading to congestive heart failure. Like MMPs, drug development is a tightly regulated process, and the successful turnover of MMP inhibitors into a marketed drug has also been a slow process on a pharmaceutical timescale. Since the discovery of the archetypal MMP (type 1 collagenase) over four decades ago by Gross and Lapierre, most major pharmaceutical companies have had MMP inhibitor programs for a variety of indications. Despite decades of research, tens of thousands of compounds synthesized and screened, and billions of dollars spent in clinical studies Periostat (doxycycline hyclate, CollaGenex Pharmaceuticals Inc.) is the only collagenase inhibitor to be successfully launched. In addition, Periostat's approval is currently limited to periodontal disease. This article focuses on some of the lessons to be learned from the failure of so many MMP inhibitors across so many indications, and what potential exists for MMP inhibitors as a drug class, especially for heart failure. PMID- 14739772 TI - [Raynaud-like phenomenon in infants with atopic dermatitis: correlation with serum endothelin-1 levels]. AB - Infants with severe atopic dermatitis (AD) sometimes have cold and pale fingers and toes as observed in patients with Raynaud-like phenomenon (RP). We tried to clarify the physiological mechanism of secondary RP in patients with AD. The correlation between serum endothelin-1 (ET-1) or nitrate (NO3) levels and the severity of AD was examined in 37 patients. As a result, RP was observed in 5 boys younger than 6 months of age and with severe AD. These 5 infants had high serum ET-1 levels. However, serum NO3 levels were only mildly elevated. These results suggest that secondary RP in AD may occur with elevated ET-1 caused by stressed and/or damaged endothelium in infants with severe AD. PMID- 14739773 TI - [Construction and operation of Internet Search Engine specialized in information on asthma. A Search Engine-based investigation to identify asthma-related information needed by Internet users]. AB - To support asthmatic patients in collecting information through the Internet, we have constructed and operated a search engine specialized in asthma-related information making use of the search engine software available free of charge and other programs. A questionnaire was attached to the bottom of the Web page presenting the search results, asking the users to respond to several questions. During the three-year period since its start of operation on June 1, 2000, there was 66689 visits to this site and 786 responses to the questionnaire were collected. Of all respondents, 19.3% were medical professionals, 63.7% were patients or their family members, and 11.3% belonged to the other categories (5.3% did not specify their position). In each of these three user groups, only about half of the users were able to find a route to the information they needed. This seems to reflect the absence of adequate asthma-related information sources on the Internet in Japanese language. However, more than 70% of all users in each group answered that this search engine site was useful. PMID- 14739774 TI - [Clinical evaluation of urinary leukotriene e4 levels in children with respiratory syncytial virus infection]. AB - The levels of leukotriene E4 (LTE4) of the urine were determined in 24 pediatric patients with infectious diseases due to respiratory syncytial virus (RSV), i.e., bronchitis, pneumonia, and bronchiolitis, and compared with those in controls without allergic disease. The level for LTE4 of the acute-phase urine was 620+/ 562 pg/mg. cr in the pediatric patients infected with RSV, being significantly higher than 190+/-67 pg/mg. cr in controls (P<0.005). The levels for LTE4 of the urine in the recovery phase showed a tendency toward decrease, as compared to those in the acute phase. However, there was no significant difference in the level for LTE4 of the acute-phase urine between the presence and the absence of each of the following conditions: expiratory wheezing; the association of pneumonia; family history of allergic diseases; the association of atopic dermatitis; and a past history of expiratory wheezing. An allergological study also revealed that there was no significant difference in LTE4 level between the presence and the absence of peripheral eosinophilia or between the presence and the absence of the high total level for IgE of the serum or positivity for the specific IgE level in the serum. These results suggest that LT is involved with the pathological conditions of RSV infection, but there are no direct relation between atopic diathesis or expiratory wheezing and the amounts of LT production. PMID- 14739775 TI - [A case of zaltoprofen induced aseptic meningitis in Sjogren's syndrome]. PMID- 14739776 TI - Pinning down transcription: regulation of RNA polymerase II activity during the cell cycle. AB - Pin1 is a peptidyl-prolyl cis/trans isomerase that was initially discovered as an important mitotic regulator. In keeping with this, it targets specifically phosphorylated ser- or thr-pro dipeptides, which are found in numerous mitotic proteins as a result of cyclin-dependent kinase activity. But Pin1 also modulates the activity of numerous other proteins, and a strong candidate for such regulation has been RNA polymerase II (RNAP II). The unique C-terminal domain of the RNAP II largest subunit contains as many as 100 ser-pro dipeptides laid out in an essentially linear array, and which are subject to different patterns of phosphorylation. Here we summarize findings indicating that Pin1 can indeed directly modulate the structure and function of RNAP II, and propose that this activity plays a role both in mitotic gene silencing and during the transcription cycle. PMID- 14739777 TI - MdmX represses E2F1 transactivation. AB - Based on knockout mouse studies, Mdm2 and MdmX have been identified as critical regulators of the p53 tumor suppressor protein, at least during early development. While many of the functions attributed to Mdm2 and MdmX involve p53 and overexpression of each gene appears to have oncogenic activities, a number of studies have suggested that each protein also possesses p53-independent functions. While examining the effect of Mdm2 overexpression on E2F1 transactivation we uncovered a novel MdmX function, the ability to inhibit E2F1 transactivation in a p53 and Mdm2 independent manner. Using a series of MdmX deletion mutants the central region of MdmX, amino acids 128-444 appears to possess the repressive domain. While an in vivo association of MdmX with either E2F1 or DP1 was not observed, a slight reduction in DP1 and an increased cytoplasmic localization of E2F1 were seen in cells overexpressing MdmX. These results suggest that elevated MdmX expression may repress E2F1-regulated genes like p14ARF and thus represent another regulatory mechanism in the Rb-p53 signaling pathway. PMID- 14739778 TI - Reassessing the usual suspects causing a commotion in the blood (and vessels?). PMID- 14739779 TI - Inhibition of Tie-2 signaling induces endothelial cell apoptosis, decreases Akt signaling, and induces endothelial cell expression of the endogenous anti angiogenic molecule, thrombospondin-1. AB - Small molecule inhibitors of endothelial cell specific tyrosine kinases are currently under investigation as potential means to block tumor angiogenesis. We have investigated the utility of blocking Tie-2 signaling in endothelial cells as a potential anti-angiogenic strategy. We have found that interruption of Tie-2 signaling either via RNAi or overexpression of a kinase-dead Tie-2 led to loss of endothelial cell viability, even in the presence of serum. Mechanistically, this is linked to a block in Akt signaling and increased thrombospondin expression. Thrombospondins are endogenous anti-angiogenic matricellular proteins known to regulate tumor growth and angiogenesis. We observed that both Tie-2 and subsequent PI3Kinase signaling regulates thrombospondin-1 expression. These data have lead to the model that Angiopoietin signaling through Tie-2 activates PI3Kinase/Akt, which represses thrombospondin expression. Thus, targeting Tie-2 with small molecules maybe efficacious as an anti-angiogenic therapy. PMID- 14739780 TI - Flow cytometric analysis of electronic nuclear volume and DNA content in normal mouse tissues. AB - Light scatter is used in flow cytometry for identification of cells based on their size and/or granularity. However, forward light scatter is not an accurate measure of cell size. The measurement of Electronic Volume (EV) by Coulter principle is more accurate. However, EV cannot be measured on most of the commercially available flow cytometers. We have described the development and applications of a flow cytometer that can simultaneously measure Electronic Nuclear Volume (ENV) and DNA content. In the present study we have used a commercially available NPE Quanta for measuring EV and DNA content of different normal mice tissues. Fresh/frozen or formalin fixed-paraffin embedded tissues from mice were processed for isolation of nuclei, which were then analyzed for EV versus DNA content. By using these two parameters, distinct sub-populations were identified in liver, thymus, small intestine and bone marrow. Dual parametric analysis of EV versus DNA content can be a valuable technique for identification of sub-populations in heterogeneous cell mixtures such as those of complex tissues like bone marrow, intestine and tumors. The methods established are rapid and can provide valuable data for identification and characterization of sub populations for cell cycle analysis by flow cytometry. PMID- 14739781 TI - Family feud in chemosensitvity: p73 and mutant p53. AB - The importance of p53 in chemotherapy-induced apoptosis of cancer cells is well established. p53 plays a critical role in the cellular response to DNA damage by regulating genes involved in cell cycle progression, apoptosis, and genomic stability. As a result, p53 tumor status is a critical determinant of both responses to anti-cancer treatment and clinical prognosis. Interestingly, tumors expressing certain mutant forms of p53 ("gain of function") are particularly resistant to chemotherapy, even when compared to cells that lack any detectable p53. Until recently, the explanation for this enhanced chemoresistance was not clear. Recent studies have shown that the p53 homologues, p73 and p63, are also activated by chemotherapies, leading to tumor cell death. Now the discovery that mutant p53 interacts with p73, and that regulation of this interaction by a p53 polymorphism can modulate chemosensitvity provide a new model for how p53-family interactions can influence the response of tumors to anti-cancer therapies. Since p53 mutations are found in more than 50% of human tumors, strategies aimed at manipulating these interactions may prove useful in enhancing the chemotherapy response, and perhaps, overcoming chemoresistance. PMID- 14739782 TI - Wnt signaling and breast cancer. AB - Secreted signaling factors of the Wnt protein family regulate many cellular processes, including cell fate decisions and cell proliferation, and aberrant Wnt signaling is associated with tumorigenesis. Many Wnt proteins act via a signaling pathway that results in stabilization of beta-catenin and consequent transcriptional activation of specific target genes. Mutations in beta-catenin or other Wnt pathway components, which result in beta-catenin accumulation, are found in a wide range of human cancers. In contrast, such mutations have been found only rarely in breast cancer. Nevertheless there is strong evidence of stabilization of beta-catenin protein in a majority of human breast tumors. Moreover, studies in mouse model systems clearly demonstrate that activated Wnt signaling leads to mammary tumorigenesis. This review summarizes the current evidence implicating Wnt/beta-catenin signaling in breast cancer and discusses several possible mechanisms by which the pathway may become activated. PMID- 14739783 TI - Targeting PAK etk... PMID- 14739784 TI - Targeting vascular and avascular compartments of tumors with C. novyi-NT and anti microtubule agents. AB - Current approaches for treating cancer are limited, in part, by the inability of drugs to affect the poorly vascularized regions of tumors. We have found that C. novyi-NT in combination with anti-microtubule agents can cause the destruction of both the vascular and avascular compartments of tumors. The two classes of microtubule inhibitors were found to exert markedly different effects. Some agents that inhibited microtubule synthesis, such as HTI-286 and vinorelbine, caused rapid, massive hemorrhagic necrosis when used in combination with C. novyi NT. In contrast, agents that stabilized microtubules, such as the taxanes docetaxel and MAC-321, resulted in slow tumor regressions that killed most neoplastic cells. Remaining cells in the poorly perfused regions of tumors could be eradicated by C. novyi-NT. Mechanistic studies showed that the microtubule destabilizers, but not the microtubule stabilizers, radically reduced blood flow to tumors, thereby enlarging the hypoxic niche in which C. novyi-NT spores could germinate. A single intravenous injection of C. novyi-NT plus selected anti microtubule agents was able to cause regressions of several human tumor xenografts in nude mice in the absence of excessive toxicity. PMID- 14739785 TI - Topotecan in small cell lung cancer: first or second line? PMID- 14739786 TI - Regulation of tumor progression by anti-neoplastic T cell responses. AB - A central paradox in tumor immunology is the seeming coexistence of a developing or progressing tumor burden in the absence of a therapeutically effective host anti-neoplastic response. It is now generally thought, however, that the host, whether mouse or human, can mount a T cell response against a developing malignancy without any prior immunization. Ostensibly, the nature or potency of that antitumor response is frequently insufficient to alter the course of neoplastic progression. Consequently, cancer immunotherapy was born from the idea that a preexisting or newly induced immune response may be intensified to achieve clinical regression. At the heart of that matter is the capacity of the host immune system to not only phenotypically distinguish normal from malignant cells, but also to functionally mediate antitumor activity. In both preclinical models and clinical settings, a plethora of tumor-associated or tumor-specific antigens have now been identified that can induce anti-neoplastic CD4+ or CD8+ T cell responses. Thus, immunization against neoplastic disease is attainable and, in several cases, overcomes potential mechanisms of immune tolerance and escape. In terms of objective clinical responses, however, the results have been less dramatic, and only a smaller proportion of patients have undergone significant disease regression. These responses have been observed mainly in clinical settings of adoptive T cell therapy, as compared with vaccination approaches. Nonetheless, these studies collectively provide the rationale for the further development and application of active and adoptive immunotherapies, perhaps in combination with each other or with oncological interventions, to promote more meaningful and sustainable anti-neoplastic immune responses and clinical outcomes. PMID- 14739788 TI - Discharge criteria for children sedated by nonanesthesiologists: is "safe" really safe enough? PMID- 14739789 TI - Long-lasting changes in brain protein expression after exposure to an anesthetic. PMID- 14739790 TI - Genetic testing for malignant hyperthermia in North America. PMID- 14739791 TI - Peter J Pronovost, M.D., Ph.D. Recipient of the 2003 Presidential Scholar Award. PMID- 14739792 TI - Can we improve the assessment of discharge readiness?: A comparative study of observational and objective measures of depth of sedation in children. AB - BACKGROUND: Current recommended discharge criteria might not be rigorous enough to detect residual sedation. This study evaluated the use of the Bispectral Index (BIS monitor), the University of Michigan Sedation Scale (UMSS; i.e., 0-4 observational scale), and a Modified Maintenance of Wakefulness Test (MMWT; visual observation of the time the child is able to stay awake) in assessing return to baseline status. METHODS: Twenty-nine children sedated for echocardiographic examination were studied. Nurses administered sedatives and monitored and discharged children according to institutional guidelines. Children were monitored with the BIS(R) throughout the study. Trained observers assigned UMSS scores every 10-15 min until revised discharge criteria were met (i.e., UMSS score of 0 or 1, MMWT duration >/= 20 min). The MMWT value was recorded at each observation following the procedure. Subsequently, blinded observers recorded average BIS values for the 5 min before each UMSS observation. RESULTS: There were moderate correlations between the BIS, MMWT, and UMSS scores (r = 0.68-0.78; P < 0.01). Revised criteria correctly identified children who were awake and alert (BIS value >/= 90) in 88% of the cases. Only 55% of the children had returned to baseline BIS values when discharged by the nurse, compared with 92% when revised criteria were met (P < 0.05). It took longer to meet revised criteria compared with standard criteria (75.3 +/- 76.2 min vs. 16.4 +/- 13.1 min; P = 0.001). CONCLUSIONS: The incorporation of specific, objective discharge criteria (i.e., UMSS score of 0 or 1, MMWT duration >/= 20 min) may ensure a status closer to baseline (BIS value >/= 90) compared with nursing judgment using standard criteria. However, such assurance may delay the discharge of sedated children. PMID- 14739793 TI - Effects of epsilon-aminocaproic acid and aprotinin on leukocyte-platelet adhesion in patients undergoing cardiac surgery. AB - BACKGROUND: The administration of aprotinin during cardiopulmonary bypass (CPB) is hypothesized to decrease activation of leukocytes and platelets and possibly reduce their adhesion. Although epsilon-aminocaproic acid (EACA) shares the ability of aprotinin to inhibit excessive plasmin activity after CPB, its effect on leukocyte and platelet activation and leukocyte-platelet (heterotypic) adhesion is largely unknown. This study was performed to determine the relative effectiveness of the antifibrinolytics, aprotinin and EACA, at reducing leukocyte and platelet activation and leukocyte-platelet conjugate formation in patients undergoing CPB. METHODS: Thirty-six patients scheduled to undergo cardiac surgery with CPB were randomized in a double-blind fashion to receive EACA, aprotinin, or saline (placebo). Markers of plasmin activity (D-dimer concentrations), platelet activation (CD62P), leukocyte activation (CD11b), and leukocyte-platelet adhesion (monocyte- and neutrophil-platelet conjugates) were measured before, during, and after CPB. RESULTS: Platelet CD62P (P-selectin), monocyte CD11b, and monocyte platelet conjugates were all significantly increased (compared with baseline) in the saline group during and after CPB. Despite equivalent reductions in D-dimer formation in patients receiving EACA (P < 0.0001) and aprotinin (P < 0.0001), decreases in platelet CD62P and monocyte CD11b expression were incomplete (not significantly different from saline control). In contrast, peak monocyte-platelet conjugate formation was significantly reduced by both EACA (P = 0.026) and aprotinin (P = 0.039) immediately after CPB. CONCLUSIONS: EACA seems to be as effective as aprotinin at reducing peak monocyte-platelet adhesion after CPB. Furthermore, inhibition of excessive plasmin activity seems to influence monocyte platelet adhesion. The findings suggest that monocyte-platelet conjugate formation may be a useful marker of monocyte and platelet activation in this clinical setting. PMID- 14739794 TI - Early onset pneumonia: risk factors and consequences in head trauma patients. AB - BACKGROUND: Early onset pneumonia occurs frequently in head trauma patients, but the potential consequences and the risk factors of this event have been poorly studied. METHODS: This prospective observational study was undertaken in the surgical intensive care unit of a university teaching hospital in Clichy, France. Head trauma patients requiring tracheal intubation for neurologic reasons and ventilation for at least 2 days were studied to assess the risk factors and the consequences of early onset pneumonia. RESULTS: During a 2-yr period, 109 head trauma patients were studied. The authors found an incidence of early onset pneumonia of 41.3%. Staphylococcus aureus was the most common bacteria involved in early onset pneumonia. Patients with early onset pneumonia had a lower worst arterial oxygen tension:fraction of inspired oxygen ratio, more fever, more arterial hypotension, and more intracranial hypertension, factors known to worsen the neurologic prognosis of head trauma patients. Nasal carriage of S. aureus on admission (odds ratio, 5.1; 95% confidence interval, 1.9-14.0), aspiration before intubation (odds ratio, 5.5; 95% confidence interval, 1.9-16.4) and barbiturate use (odds ratio, 3.9; 95% confidence interval, 1.2-12.8) were found to be independent risk factors of early onset pneumonia. CONCLUSIONS: The results suggest that early onset pneumonia leads to secondary injuries in head-injured patients. Nasal carriage of S. aureus, aspiration before intubation, and use of barbiturates are specific independent risk factors for early onset pneumonia and must be assessed to find and evaluate strategies to prevent early onset pneumonia. PMID- 14739795 TI - Mixed-effects modeling of the intrinsic ventilatory depressant potency of propofol in the non-steady state. AB - BACKGROUND: Despite the ubiquitous use of propofol for anesthesia and conscious sedation and numerous publications about its effect, a pharmacodynamic model for propofol-induced ventilatory depression in the non-steady state has not been described. To investigate propofol-induced ventilatory depression in the clinically important range (at and below the metabolic hyperbola while carbon dioxide is accumulating because of drug-induced ventilatory depression), the authors applied indirect effect modeling to Paco2 data at a fraction of inspired carbon dioxide of 0 during and after administration of propofol. METHODS: Ten volunteers underwent determination of their carbon dioxide responsiveness by a rebreathing design. The parameters of a power function were fitted to the end expiratory carbon dioxide and minute ventilation data. The volunteers then received propofol in a stepwise ascending pattern with use of a target-controlled infusion pump until significant ventilatory depression occurred (end-tidal pressure of carbon dioxide > 65 mmHg and/or imminent apnea). Thereafter, the concentration was reduced to 1 microg/ml. Propofol pharmacokinetics and the Paco2 were determined from frequent arterial blood samples. An indirect response model with Bayesian estimates of the pharmacokinetics and carbon dioxide responsiveness in the absence of drug was used to describe the Paco2 time course. Because propofol reduces oxygen requirements and carbon dioxide production, a correction factor for propofol-induced decreasing of carbon dioxide production was included. RESULTS: The following pharmacodynamic parameters were found to describe the time course of hypercapnia after administration of propofol (population mean and interindividual variability expressed as coefficients of variation): F (gain of the carbon dioxide response), 4.37 +/- 36.7%; ke0, CO2, 0.95 min-1 +/- 59.8%; baseline Paco2, 40.9 mmHg +/- 12.8%; baseline minute ventilation, 6.45 l/min +/- 36.3%; kel, CO2, 0.11 min-1 +/- 34.2%; C50,propofol, 1.33 microg/ml +/- 49.6%; gamma, 1.68 +/- 21.3%. CONCLUSION: Propofol at common clinical concentrations is a potent ventilatory depressant. An indirect response model accurately described the magnitude and time course of propofol-induced ventilatory depression. The indirect response model can be used to optimize propofol administration to reduce the risk of significant ventilatory depression. PMID- 14739796 TI - Children undergoing repeated exposures for radiation therapy do not develop tolerance to propofol: clinical and bispectral index data. AB - BACKGROUND: The purpose of this study was to apply clinical criteria and Bispectral Index monitor data for evaluating the development of tolerance to propofol in children undergoing repeated drug exposure. METHODS: Children undergoing multiple sessions of radiation therapy during anesthesia for various malignancies were given a predetermined dose of propofol at each session. Heart rate, blood pressure, oxygen saturation, respiratory rate, requirement of additional propofol, and time to emergence and discharge were recorded. The Bispectral Index was monitored continuously, and parameters were extracted and averaged for each week of therapy. RESULTS: Fifteen children (aged 2.5-10 yr) were treated for an average of 5 weeks (24 +/- 6 sessions). There were no significant differences in physiologic parameters or requirements of additional propofol between the weeks of treatment. Bispectral Index data analysis showed that although a nonlinear change with time for each parameter could not be rejected, the differences between the first and last intervals were nonsignificant. CONCLUSIONS: Overall changes with time resulted from random fluctuations without a consistent trend. Combined with clinical data, Bispectral Index parameters showed that tolerance to propofol does not develop in children undergoing repeated exposures to the drug during radiation therapy. PMID- 14739797 TI - Effects of two target-controlled concentrations (1 and 3 ng/ml) of remifentanil on MAC(BAR) of sevoflurane. AB - BACKGROUND: The aim of this prospective, randomized, double-blind study was to determine the effects of two different target-controlled concentrations of remifentanil (1 and 3 ng/ml) on the sevoflurane requirement for blunting sympathetic responses after surgical incision (MACBAR). METHODS: Seventy-four patients aged 20-50 yr, with American Society of Anesthesiologists physical status I, were anesthetized with propofol, cisatracurium, and sevoflurane with a mixture of 60% nitrous oxide in oxygen. Then, patients were randomly allocated to receive no remifentanil infusion (n = 27) or a target-controlled plasma concentration of 1 ng/ml (n = 27) or 3 ng/ml remifentanil (n = 20). Sympathetic responses to surgical incision (presence or absence of an increase in either heart rate or mean arterial blood pressure of 15% or more above the mean of the values measured during the 2 min before skin incision) were determined after a 20 min period of stable end-tidal sevoflurane and target-controlled remifentanil concentrations. Predetermined end-tidal sevoflurane concentrations and the MACBAR for each group were determined using an up-and-down sequential-allocation technique. RESULTS: The MACBAR of sevoflurane was higher in the group receiving no remifentanil (2.8% [95% confidence interval: 2.5-3.0%]) as compared with patients of the groups receiving 1 ng/ml (1.1% [0.9-1.3%]; P = 0.012) and 3 ng/ml remifentanil (0.2% [0.1-0.3%]; P = 0.006). When considering a minimum anesthetic concentration (MAC) value in this age population and the contribution of 60% nitrous oxide (0.55 MAC), the combined MACBAR values, expressed as multiples of the MAC, were 1.95 MAC, 1.1 MAC, and 0.68 MAC, in the three groups, respectively. CONCLUSION: A target-controlled concentration of 1 ng/ml remifentanil results in a 60% decrease in the MACBAR of sevoflurane combined with 60% nitrous oxide. Increasing the target concentration of remifentanil to 3 ng/ml produces a further 30% decrease in the MACBAR values of sevoflurane. PMID- 14739798 TI - Antichemical protective gear prolongs time to successful airway management: a randomized, crossover study in humans. AB - BACKGROUND: Airway management is the first step in resuscitation. The extraordinary conditions in mass casualty situations impose special difficulties in airway management, even for experienced caregivers. The authors evaluated whether wearing surgical attire or antichemical protective gear made any difference in anesthetists' success of airway control with either an endotracheal tube or a laryngeal mask airway. METHODS: Fifteen anesthetists with 2-5 yr of residency and wearing either full antichemical protective gear or surgical attire intubated or inserted laryngeal masks in 60 anesthetized patients. The study was performed in a prospective, randomized, crossover manner. The duration of intubation/insertion was measured from the time the device was grasped to the time a normal capnography recording was obtained. RESULTS: Endotracheal tubes were introduced significantly (P < 0.01) faster when the anesthetist wore surgical attire (31 +/- 7 vs. 54 +/- 24 s for protective gear), but the mean times necessary to successfully insert laryngeal masks were similar (44 +/- 20 s for surgical attire vs. 39 +/- 11 s for protective gear). Neither performance failure nor incidences of hypoxemia were recorded. CONCLUSIONS: This first report in humans shows to what extent anesthetists' wearing of antichemical protective gear slows the time to intubate but not to insert a laryngeal mask airway compared with wearing surgical attire. Laryngeal mask airway insertion is faster than tracheal intubation when wearing protective gear, indicating its advantage for airway management when anesthetists wear antichemical protective gear. If chances for rapid and successful tracheal intubation under such chaotic conditions are poor, laryngeal mask airway insertion is a viable choice for airway management until a proper secured airway is obtainable. PMID- 14739799 TI - Laryngeal mask airway insertion by anesthetists and nonanesthetists wearing unconventional protective gear: a prospective, randomized, crossover study in humans. AB - BACKGROUND: Mass casualty situations impose special difficulties in airway management, even for experienced caregivers. The laryngeal mask airway is part of the difficult airway algorithm. The authors evaluated the success rate and the time to secure airways by mask by anesthetists, surgeons, and novices when wearing either surgical attire or full antichemical protective gear that included butyl rubber gloves and a filtering antigas mask. METHODS: Twenty anesthetists and 22 surgeons with 2-5 yr of residency inserted a laryngeal mask airway in 84 anesthetized patients, and 6 novices repetitively inserted masks in 57 patients under both conditions in a prospective, randomized, crossover manner. The duration of insertion was measured from the time the device was first grasped until a normal capnography recording was obtained. RESULTS: Anesthetists needed 39 +/- 14 s to insert the masks when wearing surgical attire and 40 +/- 12 s with protective gear. In contrast, surgery residents needed 64 +/- 40 and 102 +/- 40 s (P = 0.0001), respectively. Anesthetists inserted masks in a single attempt, whereas the surgeons needed up to four attempts with no hypoxia or failure associated. The initial attire-wearing novices' insertions took as long as the surgeons'; three of them then reached the mean performance time of the anesthetists after four (protective gear) and two (surgical attire) trials, with only one occurrence of hypoxia and a failure rate similar to that of the surgeons. CONCLUSIONS: Anesthesia residents insert laryngeal mask airways at a similar speed when wearing surgical attire or limiting antichemical protective gear and two to three times faster than surgical residents or novices wearing either outfit. Novices initially perform at the level of surgical residents, but their learning curve was quick under both conditions. PMID- 14739800 TI - Obesity decreases perioperative tissue oxygenation. AB - BACKGROUND: Obesity is an important risk factor for surgical site infections. The incidence of surgical wound infections is directly related to tissue perfusion and oxygenation. Fat tissue mass expands without a concomitant increase in blood flow per cell, which might result in a relative hypoperfusion with decreased tissue oxygenation. Consequently, the authors tested the hypotheses that perioperative tissue oxygen tension is reduced in obese surgical patients. Furthermore, they compared the effect of supplemental oxygen administration on tissue oxygenation in obese and nonobese patients. METHODS: Forty-six patients undergoing major abdominal surgery were assigned to one of two groups according to their body mass index: body mass index less than 30 kg/m2 (nonobese) or 30 kg/m2 or greater (obese). Intraoperative oxygen administration was adjusted to arterial oxygen tensions of approximately 150 mmHg and approximately 300 mmHg in random order. Anesthesia technique and perioperative fluid management were standardized. Subcutaneous tissue oxygen tension was measured with a polarographic electrode positioned within a subcutaneous tonometer in the lateral upper arm during surgery, in the recovery room, and on the first postoperative day. Postoperative tissue oxygen was also measured adjacent to the wound. Data were compared with unpaired two-tailed t tests and Wilcoxon rank sum test; P < 0.05 was considered statistically significant. RESULTS: Intraoperative subcutaneous tissue oxygen tension was significantly less in the obese patients at baseline (36 vs. 57 mmHg; P = 0.002) and with supplemental oxygen administration (47 vs. 76 mmHg; P = 0.014). Immediate postoperative tissue oxygen tension was also significantly less in subcutaneous tissue of the upper arm (43 vs. 54 mmHg; P = 0.011) as well as near the incision (42 vs. 62 mmHg; P = 0.012) in obese patients. In contrast, tissue oxygen tension was comparable in each group on the first postoperative morning. CONCLUSION: Wound and tissue hypoxia were common in obese patients in the perioperative period and most pronounced during surgery. Even with supplemental oxygen tissue, oxygen tension in obese patients was reduced to levels that are associated with a substantial increase in infection risk. PMID- 14739801 TI - Epidural anesthesia, hypotension, and changes in intravascular volume. AB - BACKGROUND: The most common side effect of epidural or spinal anesthesia is hypotension with functional hypovolemia prompting fluid infusions or administration of vasopressors. Short-term studies (20 min) in patients undergoing lumbar epidural anesthesia suggest that plasma volume may increase when hypotension is present, which may have implications for the choice of treatment of hypotension. However, no long-term information or measurements of plasma volumes with or without hypotension after epidural anesthesia are available. METHODS: In 12 healthy volunteers, the authors assessed plasma (125I albumin) and erythrocyte (51Cr-EDTA) volumes before and 90 min after administration of 10 ml bupivacaine, 0.5%, via a thoracic epidural catheter (T7 T10). After 90 min (t = 90), subjects were randomized to administration of fluid (7 ml/kg hydroxyethyl starch) or a vasopressor (0.2 mg/kg ephedrine), and 40 min later (t = 130), plasma and erythrocyte volumes were measured. At the same time points, mean corpuscular volume and hematocrit were measured. Systolic and diastolic blood pressure, heart rate, and hemoglobin were measured every 5 min throughout the study. Volume kinetic analysis was performed for the volunteers receiving hydroxyethyl starch. RESULTS: Plasma volume did not change per se after thoracic epidural anesthesia despite a decrease in blood pressure. Plasma volume increased with fluid administration but remained unchanged with vasopressors despite that both treatments had similar hemodynamic effects. Hemoglobin concentrations were not significantly altered by the epidural blockade or ephedrine administration but decreased significantly after hydroxyethyl starch administration. Volume kinetic analysis showed that the infused fluid expanded a rather small volume, approximately 1.5 l. The elimination constant was 56 ml/min. CONCLUSIONS: Thoracic epidural anesthesia per se does not lead to changes in blood volumes despite a reduction in blood pressure. When fluid is infused, there is a dilution, and the fluid initially seems to be located centrally. Because administration of hydroxyethyl starch and ephedrine has similar hemodynamic effects, the latter may be preferred in patients with cardiopulmonary diseases in which perioperative fluid overload is undesirable. PMID- 14739802 TI - Wrist hyperextension leads to median nerve conduction block: implications for intra-arterial catheter placement. AB - BACKGROUND: It is common practice to hyperextend the wrist to facilitate insertion of a radial intra-arterial catheter. This position may be maintained for prolonged periods. Although there has been much discussion about optimal patient management to protect the ulnar nerve and brachial plexus, little attention has been paid to the median nerve during wrist hyperextension. The authors report the effects of wrist hyperextension on conduction in the median nerve. METHODS: Median nerve conduction was studied in 12 awake, healthy volunteers using standard nerve conduction tests consisting of the measurement of compound sensory and motor action potentials, as well as their amplitudes and latencies. With the contralateral hand as a control, the right wrist was placed in hyperextension (angled between 65 and 80 degrees), and compound action potentials were recorded to determine the onset and magnitude of effects. Subsequently, the hand was released from hyperextension and recovery was recorded. RESULTS: In 83% of subjects, hyperextension resulted in a significant decrease in compound sensory action potential amplitudes, sufficient to qualify as conduction block (16.6% of baseline). The average time to conduction block was 43 +/- 13.2 min. All subjects who manifested conduction block showed marked improvement 5 min after release from hyperextension. CONCLUSIONS: Wrist hyperextension for arterial line placement and stabilization is likely to result in profound impairment of median nerve function. Although the effects were transient in this study, the results suggest that prolonged hyperextension may be associated with significant changes in median nerve conduction. To minimize the chance for nerve injury, the authors recommend that wrists be returned promptly to the neutral position following arterial line placement. PMID- 14739803 TI - An investigation to dissociate the analgesic and anesthetic properties of ketamine using functional magnetic resonance imaging. AB - BACKGROUND: Anatomic sites within the brain, which activate in response to noxious stimuli, can be identified with the use of functional magnetic resonance imaging. The aim of this study was to determine whether the analgesic effects of ketamine could be imaged. METHODS: Ketamine was administered to eight healthy volunteers with use of a target-controlled infusion to three predicted plasma concentrations: 0 (saline), 50 (subanalgesic), and 200 ng/ml (analgesic, subanesthetic). Volunteers received noxious thermal stimuli and auditory stimuli and performed a motor task within a 3-T human brain imaging magnet. Activation of brain regions in response to noxious and auditory stimuli and during the motor task was compared with behavioral measures. RESULTS: The analgesic subanesthetic dose of ketamine significantly reduced the pain scores, and this matched a decrease in activity within brain regions that activate in response to noxious stimuli, in particular, the insular cortex and thalamus. A different pattern of activation was observed in response to an auditory task. In comparison, smaller behavioral and imaging changes were found for the motor paradigm. The lower dose of ketamine gave similar but smaller nonsignificant effects. CONCLUSION: The analgesic effect can be measured within a more global effect of ketamine as shown by auditory and motor tasks, and the analgesia produced by ketamine occurs with a smaller degree of cortical processing in pain-related regions. PMID- 14739804 TI - Alterations in rat brain proteins after desflurane anesthesia. AB - BACKGROUND: Volatile anesthetics disappear from an organism after the end of anesthesia. Whether changes of protein expression persist in the brain for a longer period is not known. This study investigates the question of whether the expression of proteins is altered in the rat brain after the end of desflurane anesthesia. METHODS: Three groups (n = 12 each) of rats were anesthetized with 5.7% desflurane in air for 3 h. Brains were removed directly after anesthesia, 24 h after anesthesia, or 72 h after anesthesia. Two additional groups (n = 12 each) served as naive conscious controls, in which the brains were removed without previous anesthesia 3 or 72 h after the start of the experiment. Cytosolic proteins were isolated. A proteome-wide study was performed, based on two dimensional gel electrophoresis and mass spectrometry. RESULTS: Compared with conscious controls, significant (P < 0.05) increase/decrease was found: 3 h of anesthesia, 5/2 proteins; 24 h after anesthesia, 13/1 proteins; 72 h after anesthesia, 6/4 proteins. The overall changes in protein expression as quantified by the induction factor ranged from -1.67 (decrease to 60%) to 1.79 (increase by 79%) compared with the controls (100%). Some of these regulated proteins play a role in vesicle transport and metabolism. CONCLUSION: Desflurane anesthesia produces changes in cytosolic protein expression up to 72 h after anesthesia in the rat brain, indicating yet unknown persisting effects. PMID- 14739805 TI - Long-term impairment of acquisition of a spatial memory task following isoflurane nitrous oxide anesthesia in rats. AB - BACKGROUND: The authors demonstrated previously that isoflurane-nitrous oxide anesthesia attenuates performance improvement on an already-learned spatial memory task and that the effect persists for weeks. This experiment was designed to test the hypothesis that learning of new information is particularly susceptible to prolonged disruption after general anesthesia. METHODS: Six- (n = 5) and 20- (n = 5) month-old male Fischer 344 rats were anesthetized for 2 h with 1.2% isoflurane, 70% nitrous oxide, and 30% oxygen. Age-matched control rats received 30% oxygen and 70% nitrogen (n = 5 per group). Rats breathed spontaneously, and anesthetic and oxygen concentrations were measured. Spatial learning was assessed daily for 21 days on a 12-arm radial maze (RAM) beginning 48 h after anesthesia. In a post hoc experiment to examine locomotion, swim speed was assessed in a separate group of identically treated rats (n = 3 per group) for 4 days beginning 48 h after anesthesia. RESULTS: Aged rats were slower to complete the maze, made fewer correct choices before first error, and made more errors at baseline than young rats (P < 0.05). Anesthesia worsened maze performance in both age groups, as evidenced by increased time to complete the maze and a decreased number of correct choices before first error (P < 0.05), but there were no statistically significant differences in total number of errors. Interestingly, there were no age-by-anesthesia interactions. Aged rats swam slower than adult rats (P < 0.001), but there were no differences between the control and anesthesia groups. CONCLUSIONS: Isoflurane-nitrous oxide anesthesia is associated with a persistent deficit in RAM performance that is not explained by impaired locomotion. This impairment occurs in adult and aged rats, indicating that it is not an age-specific phenomenon. Thus, RAM performance is altered after general anesthesia for longer than predicted by the pharmacology of the drugs used, which, by inference, suggests a long-term deficit in learning/memory. PMID- 14739806 TI - Pharmacokinetics of intrathecal oligodeoxynucleotides. AB - BACKGROUND: Intrathecal administration of antisense oligonucleotides is a frequently used technique to alter gene expression for research purposes. However, in the future, antisense oligonucleotides will likely be administered intrathecally to humans for therapeutic purposes. To date, there have been no systematic studies of the pharmacokinetics of intrathecal oligonucleotides. This study was designed to fill that knowledge gap. METHODS: Microdialysis probes were placed intrathecally at the L4, L1, and T11 vertebral levels and epidurally at the L4 vertebral level in pigs. One of the study oligodeoxynucleotides (10-, 18-, or 30-nucleotide-long sequences of the human MDR-1 gene) was injected intrathecally at the L4 level at time 0. Microdialysis samples were obtained for measurement of oligodeoxynucleotide samples at 5-min intervals until 20 min, 10 min intervals until 60 min, and 20-min intervals until 180 min. Noncompartmental pharmacokinetic analysis was performed using PK Solutions software. RESULTS: Mean residence time and terminal elimination half-life did not differ significantly among the three oligodeoxynucleotides at any sampling site. In contrast, area under the concentration-time curve differed significantly among the oligodeoxynucleotides at all sampling sites and was inversely related to oligodeoxynucleotide length at the L4 and L1 intrathecal sites but not the T11 or epidural sampling sites. Similarly, clearance and volumes of distribution at the L4 level differed significantly among the oligodeoxynucleotides and were directly related to oligodeoxynucleotide length. CONCLUSION: The intrathecal pharmacokinetics of oligodeoxynucleotides are largely determined by oligodeoxynucleotide length. This contrasts with smaller drug molecules, such as opioids, for which intrathecal and epidural pharmacokinetics are largely determined by lipid solubility, not size. The potential clinical utility of this information is that oligodeoxynucleotide distribution within the central nervous system may be controllable to some degree by varying oligodeoxynucleotide length. PMID- 14739807 TI - Mechanism by which a sustained inflation can worsen oxygenation in acute lung injury. AB - BACKGROUND: Sustained lung inflations (recruitment maneuvers [RMs]) are occasionally used during mechanical ventilation of patients with acute lung injury to restore aeration to atelectatic alveoli. However, RMs do not improve, and may even worsen, gas exchange in a fraction of these patients. In this study, the authors sought to determine the mechanism by which an RM can impair gas exchange in acute lung injury. METHODS: The authors selected a model of acute lung injury that was unlikely to exhibit sustained recruitment in response to a lung inflation. In five sheep, lung injury was induced by lavage with 0.2% polysorbate 80 in saline. Positron emission tomography and [13N]nitrogen were used to assess regional lung function in dependent, middle, and nondependent lung regions. Physiologic data and positron emission scans were collected before and 5 min after a sustained inflation (continuous positive airway pressure of 50 cm H2O for 30 s). RESULTS: All animals showed greater loss of aeration and higher perfusion and shunting blood flow in the dependent region. After the RM, Pao2 decreased in all animals by 35 +/- 22 mmHg (P < 0.05). This decrease in Pao2 was associated with redistribution of pulmonary blood flow from the middle, more aerated region to the dependent, less aerated region (P < 0.05) and with an increase in the fraction of pulmonary blood flow that was shunted in the dependent region (P < 0.05). Neither respiratory compliance nor aeration of the dependent region improved after the RM. CONCLUSIONS: When a sustained inflation does not restore aeration to atelectatic regions, it can worsen oxygenation by increasing the fraction of pulmonary blood flow that is shunted in nonaerated regions. PMID- 14739808 TI - Hypothermic preconditioning increases survival of purkinje neurons in rat cerebellar slices after an in vitro simulated ischemia. AB - BACKGROUND: A period of hypothermia before ischemia (hypothermic preconditioning) induces a delayed phase of ischemic tolerance in rat brain. However, whether hypothermic preconditioning induces an acute phase (within a few hours after the hypothermia) of ischemic tolerance remains unknown. This study was designed to determine the time window of the hypothermic preconditioning-induced acute phase of neuroprotection, which is useful information for situations during surgery with anticipated ischemic episodes, and its involved mechanisms. METHODS: Survival of Purkinje cells in rat cerebellar slices was evaluated after a 20-min oxygen-glucose deprivation (OGD, in vitro simulated ischemia) followed by a 4-h recovery. Mild hypothermia (33 degrees C) for 20 min was applied at various times before the OGD. RESULTS: The hypothermia applied immediately to 3 h before the OGD equally effectively reduced OGD-induced Purkinje cell death/injury. Glibenclamide, a selective KATP channel blocker; 8-cyclopentyl-1,3 dipropylxanthine, a selective adenosine A1 receptor antagonist; and farnesyl protein transferase inhibitor III, a selective inhibitor to reduce Ras farnesylation, abolished hypothermic preconditioning-induced neuroprotection when applied during the hypothermia. OGD increased the expression of high-mobility group I(Y) proteins, which are nuclear transcription factors to enhance the expression of putatively damaging proteins such as cyclooxygenase-2, in cerebellar slices. This increase was attenuated by hypothermic preconditioning. CONCLUSIONS: Hypothermic preconditioning induces an acute phase of neuroprotection. This neuroprotection depends on activation of the signaling molecules, adenosine A1 receptors, KATP channels, and Ras. Inhibition of putatively damaging proteins via the effects of hypothermic preconditioning on high-mobility group I(Y) expression may also be involved in hypothermic preconditioning-induced neuroprotection. PMID- 14739809 TI - Molecular mechanisms of the inhibitory effects of propofol and thiamylal on sarcolemmal adenosine triphosphate-sensitive potassium channels. AB - BACKGROUND: Both propofol and thiamylal inhibit adenosine triphosphate-sensitive potassium (KATP) channels. In the current study, the authors investigated the effects of these anesthetics on the activity of recombinant sarcolemmal KATP channels encoded by inwardly rectifying potassium channel (Kir6.1 or Kir6.2) genes and sulfonylurea receptor (SUR1, SUR2A, or SUR2B) genes. METHODS: The authors used inside-out patch clamp configurations to investigate the effects of propofol and thiamylal on the activity of recombinant KATP channels using COS-7 cells transfected with various types of KATP channel subunits. RESULTS: Propofol inhibited the activities of the SUR1/Kir6.2 (EC50 = 77 microm), SUR2A/Kir6.2 (EC50 = 72 microm), and SUR2B/Kir6.2 (EC50 = 71 microm) channels but had no significant effects on the SUR2B/Kir6.1 channels. Propofol inhibited the truncated isoform of Kir6.2 (Kir6.2DeltaC36) channels (EC50 = 78 microm) that can form functional KATP channels in the absence of SUR molecules. Furthermore, the authors identified two distinct mutations R31E (arginine residue at position 31 to glutamic acid) and K185Q (lysine residue at position 185 to glutamine) of the Kir6.2DeltaC36 channel that significantly reduce the inhibition of propofol. In contrast, thiamylal inhibited the SUR1/Kir6.2 (EC50 = 541 microm), SUR2A/Kir6.2 (EC50 = 248 microm), SUR2B/Kir6.2 (EC50 = 183 microm), SUR2B/Kir6.1 (EC50 = 170 microm), and Kir6.2DeltaC36 channels (EC50 = 719 microm). None of the mutants significantly affects the sensitivity of thiamylal. CONCLUSIONS: These results suggest that the major effects of both propofol and thiamylal on KATP channel activity are mediated via the Kir6.2 subunit. Site-directed mutagenesis study suggests that propofol and thiamylal may influence Kir6.2 activity by different molecular mechanisms; in thiamylal, the SUR subunit seems to modulate anesthetic sensitivity. PMID- 14739810 TI - Determinants of the sensitivity of AMPA receptors to xenon. AB - BACKGROUND: There is substantial and growing literature on the actions of general anesthetics on a variety of neurotransmitter-gated ion channels, with the greatest attention being focused on inhibitory gamma-amino butyric acid type A receptors. In contrast, glutamate receptors, the most important class of fast excitatory neurotransmitter-gated receptor channels, have received much less attention, and their role in the production of the anesthetic state remains controversial. METHODS: alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors formed from a variety of different subunits were expressed in Xenopus oocytes and HEK-293 cells, and their sensitivities to the inhalational general anesthetics xenon, isoflurane, and halothane were determined using two electrode voltage clamp and patch clamp techniques. The effects of desensitization on anesthetic sensitivity were investigated using cyclothiazide and site-directed mutagenesis. An ultrarapid application system was also used to mimic rapid high-concentration glutamate release at synapses. RESULTS: The authors show that xenon can potently inhibit AMPA receptors when assayed using bath application of kainate. However, when the natural neurotransmitter l glutamate is used under conditions in which the receptor desensitization is blocked and the peak of the glutamate-activated response can be accurately measured, the pattern of inhibition changes markedly. When desensitization is abolished by a single-point mutation (L497Y in GluR1 and the equivalent mutation L505Y in GluR4), the xenon inhibition is eliminated. When AMPA receptors are activated by glutamate using an ultrarapid application system that mimics synaptic conditions, sensitivity to xenon, halothane, and isoflurane is negligible. CONCLUSIONS: AMPA receptors, when assayed in heterologous expression systems, showed a sensitivity to inhalational anesthetics that was minimal when glutamate was applied rapidly at high concentrations. Because these are the conditions that are most relevant to synaptic transmission, the authors conclude that AMPA receptors are unlikely to play a major role in the production of the anesthetic state by inhalational agents. PMID- 14739811 TI - Effects of the alpha2-adrenoceptor agonist dexmedetomidine on bronchoconstriction in dogs. AB - BACKGROUND: Tracheal intubation can elicit reflex bronchoconstriction in patients with asthma or chronic obstructive pulmonary disease, complicating mechanical ventilation and weaning from mechanical support. In vitro studies of human and animal bronchial tissue indicate that alpha2-adrenoceptor stimulation can lead to smooth muscle relaxation and prevention of bronchoconstriction. Dexmedetomidine is a selective alpha2-adrenoceptor agonist approved for sedation in the intensive care unit. Whether dexmedetomidine can affect reflex bronchoconstriction is unknown. METHODS: After the approval of the institutional animal care and use committee, five mongrel dogs were anesthetized with thiopental, endotracheally intubated, and ventilated, and their airways were challenged with histamine. High resolution computed tomography was used to measure airway luminal areas at baseline and after nebulized histamine. After recovery to baseline, on separate days, dexmedetomidine (0.5 microg/kg) was administered either intravenously or as an aerosol, and the histamine challenge was repeated. RESULTS: At baseline, histamine constricted the airways to 66 +/- 27% (mean +/- SD) (P < 0.0001) and 59 +/- 30% (P < 0.0001) of maximum on the days dexmedetomidine was administered by intravenous and inhalational means, respectively. After recovery, intravenous administration of dexmedetomidine blocked the histamine-induced bronchoconstriction (87 +/- 30.4% of maximum, compared with histamine alone (P < 0.0001), whereas dexmedetomidine administered by inhalation showed no protective effect (45 +/- 30% of maximum; P < 0.0001 compared with histamine alone). CONCLUSION: alpha2-Adrenoceptor stimulation with intravenous dexmedetomidine completely blocked histamine-induced bronchoconstriction in dogs. Therefore, dexmedetomidine might be beneficial to decrease airway reactivity in patients with chronic obstructive pulmonary disease or asthma, particularly during weaning from mechanical ventilation, when neurally mediated airway reflexes may be elicited. PMID- 14739812 TI - Upregulation of spinal cyclooxygenase-2 in rats after surgical incision. AB - BACKGROUND: Although upregulation of cyclooxygenase (COX)-2 in spinal cord after peripheral inflammation has been well documented, the effect of surgery on spinal COX-2 has not been examined in detail. The present study uses a bilateral foot incision in rats to examine the magnitude and duration of surgically induced changes in spinal COX-2 protein. METHODS: A longitudinal incision was made in both plantar hind paws of isoflurane-anesthetized rats. Spinal cords were removed at various postoperative times (1-48 h), and spinal COX-2 protein levels were compared with the results of Western blot analysis. Ropivacaine-induced blockade of sciatic nerve function was used to determine the importance of afferent nerve activity on spinal COX-2 after incision. Dexamethasone and the COX-2-selective inhibitor L-745,337 were administered intrathecally to modulate spinal COX-2 after incision. RESULTS: COX-2 protein levels increased in the lumbar spinal cord at 3 (1.32-fold) and 6 (1.26-fold) h after bilateral foot incision. At later times, lumbar COX-2 levels were no different than in control animals not undergoing surgery. Cervical COX-2 protein levels remained unchanged. Sciatic nerve blockade with ropivacaine did not prevent the increase in lumbar spinal COX 2 protein levels after incision. Intrathecal dexamethasone decreased lumbar spinal COX-2 levels after incision, and an intrathecal COX-2-selective inhibitor did not reduce the COX-2 upregulation. CONCLUSIONS: After bilateral foot incision in rats, lumbar spinal COX-2 protein levels increase, although the magnitude and duration are less than reported in models of peripheral inflammation. This COX-2 upregulation does not seem to be mediated by afferent nerve activity. PMID- 14739813 TI - Ophthalmic regional anesthesia: medial canthus episcleral (sub-tenon) single injection block. AB - BACKGROUND: The purpose of this study was to evaluate the efficacy and safety of episcleral single-injection anesthesia in a large number of patients. METHODS: Over a period of 5 yr, in four institutions, anesthesiologists involved in this prospective study completed a standardized form to evaluate single-injection medial canthus high-volume episcleral anesthesia. The success rate of the block was rated according to an akinesia score. The study parameters included demographic data, surgical procedure, and anesthetic management. All patients were followed up at least until postoperative day 1, and all complications, pain, and discomfort were noted. Statistical analysis was done to assess the risk factors for complications. RESULTS: A total of 2,031 patients were included in the study. The most frequent surgical procedures performed were phacoemulsification and posterior chamber artificial lens implantation (91.0%). A total of 66 complications (3.3%) occurred in 60 patients. One patient had a retrobulbar hemorrhage, and 59 had one or two more minor incidents or pain/discomfort with the procedure. The complications consisted of subconjunctival hematoma (1.3%), ocular hypertonia (0.4%), and chemosis (0.30%). Statistical analysis revealed that inexperience in the technique represented a risk factor for complications. CONCLUSIONS: This is the first survey of a large experience in episcleral single-injection anesthesia, a form of anesthesia that does not preclude sharp-needle complications and does require training. Only one complication occurred among 2,031 patients; however, a larger number of patients is needed to definitively evaluate the safety of episcleral single-injection anesthesia. PMID- 14739814 TI - Paravertebral analgesia with levobupivacaine increases postoperative flap tissue oxygen tension after immediate latissimus dorsi breast reconstruction compared with intravenous opioid analgesia. AB - BACKGROUND: Directly measured tissue oxygen tension (Pto2) reflects the adequacy of local tissue oxygenation and influences surgical wound healing. Epidural analgesia increases Pto2 compared with intravenous morphine analgesia after abdominal surgery. The authors tested the hypothesis that paravertebral regional anesthesia and analgesia would increase Pto2 compared with intravenous opioid based anesthesia and analgesia. METHODS: Twenty patients scheduled to undergo mastectomy with immediate latissimus dorsi breast reconstruction were randomized to receive either general anesthesia with postoperative intravenous morphine analgesia or combined general-paravertebral anesthesia with continuous paravertebral postoperative analgesia using levobupivacaine in this prospective, cohort study. All patients had a local tissue oxygen sensor implanted in the flap muscle. Data were downloaded continuously for 20 h postoperatively. RESULTS: The mean Pto2 over the 20-h period was significantly higher in patients receiving paravertebral anesthesia (75 +/- 38 vs. 44 +/- 23 mmHg [mean +/- SD]; P = 0.03). Intraoperative blood loss was less in paravertebral patients (1.2 +/- 0.4 vs. 1.7 +/- 0.5 l; P = 0.04). Dynamic visual analog scale pain scores were significantly lower in paravertebral patients. Intraoperative and postoperative fluids administered, hemoglobin, core temperature, intraoperative end-tidal carbon dioxide, and mean arterial pressure were similar in both groups. CONCLUSION: The postoperative latissimus dorsi flap Pto2 was higher for 20 h after breast reconstruction with paravertebral analgesia compared with intravenous morphine analgesia. PMID- 14739816 TI - Dose-dependent effects of propofol on the central processing of thermal pain. AB - BACKGROUND: Anatomic and physiologic data show that multiple regions of the forebrain are activated by pain. However, the effect of anesthetic level on nociceptive input to these regions is not well understood. METHODS: The authors used positron emission tomography to measure the effect of various concentrations of propofol on pain-evoked changes in regional cerebral blood flow. Fifteen volunteers were scanned while warm and painful heat stimuli were presented to the volar forearm using a contact thermode during administration of target propofol concentrations of 0.0 microg/ml (alert control), 0.5 microg/ml (mild sedation), 1.5 microg/ml (moderate sedation), and 3.5 microg/ml (unconsciousness). RESULTS: During the 0.5-microg/ml target propofol concentration (mild sedation), the subjects' pain ratings increased relative to the alert control condition; correspondingly, pain-evoked regional cerebral blood flow increased in the thalamus and the anterior cingulate cortex. In contrast, when subjects lost consciousness (3.5 microg/ml), pain-evoked responses in the thalamus and the anterior cingulate cortex were no longer observed, whereas significant pain evoked activation remained in the insular cortex. CONCLUSION: These data show that propofol has a dose-dependent effect on thalamocortical transfer of nociceptive information but that some pain-evoked cortical activity remains after loss of consciousness. PMID- 14739815 TI - Epidural neostigmine produces analgesia but also sedation in women after cesarean delivery. AB - BACKGROUND: Intrathecal neostigmine produces analgesia but also nausea, limiting its utility. In contrast, epidural administration of neostigmine has been suggested to produce postoperative analgesia without nausea in nonpregnant patients. The purpose of this study was to examine the dose range for efficacy and side effects of epidural neostigmine in women at cesarean delivery receiving combined spinal-epidural anesthesia. METHODS: After institutional approval and informed consent, 80 patients for elective cesarean delivery were given combined spinal-epidural anesthesia with 8 mg hyperbaric bupivacaine plus 10 microg fentanyl. Patients were randomized to receive either saline or 75, 150, or 300 microg neostigmine (n = 20 per group) in 10 ml saline after cord clamping. Pain, morphine consumption, and side effects were monitored for 24 h. RESULTS: Global pain assessment for the first 24 h was reduced from 5.4 +/- 0.2 in the saline group to 3.0-3.5 +/- 0.3 in the neostigmine groups, dose independently. Correspondingly, global satisfaction with neostigmine was also improved (P < 0.05). Nausea and morphine consumption were similar among groups. Intraoperative shivering and sedation were increased in the 300-microg neostigmine group only (P < 0.05), and postoperative sedation was increased by neostigmine in a dose independent fashion (P < 0.05). CONCLUSIONS: Epidural neostigmine produced modest analgesia in women after cesarean delivery. In contrast with previous reports, which focused primarily on nausea, these data suggest that epidural neostigmine can also produce mild sedation for several hours. These data suggest a limited role for single bolus-administration epidural neostigmine for analgesia after cesarean delivery. They also support future study of epidural neostigmine for obstetric analgesia. PMID- 14739817 TI - Infraclavicular perineural local anesthetic infusion: a comparison of three dosing regimens for postoperative analgesia. AB - BACKGROUND: In this randomized, double-blind study, the authors investigated the efficacy of continuous and patient-controlled ropivacaine infusions via an infraclavicular perineural catheter in ambulatory patients undergoing moderately painful orthopedic surgery at or distal to the elbow. METHODS: Preoperatively, patients (n = 30) received an infraclavicular perineural catheter and nerve block. Postoperatively, patients were discharged home with both oral analgesics and a portable infusion pump delivering 0.2% ropivacaine (500-ml reservoir) in one of three dosing regimens: the basal group (12 ml/h basal, 0.05-ml patient controlled bolus dose), the basal-bolus group (8 ml/h basal, 4 ml bolus), or the bolus group (0.3 ml/h basal, 9.9 ml bolus). Investigators and patients were blinded to random group assignment. RESULTS: The basal group (n = 10) required more oral analgesics than the basal-bolus group (P = 0.002) and had a shorter median infusion duration than the other two groups (P < 0.001 for both). The bolus group had the longest median infusion duration (P < 0.001 for both) but experienced an increase in breakthrough pain incidence (P = 0.004) and intensity (P = 0.04 vs. basal-bolus group) as well as sleep disturbances (P < 0.001 for both) compared with the other groups. Overall satisfaction was greatest in the basal-bolus group (9.7 +/- 0.5 vs. 7.9 +/- 1.7 and 8.1 +/- 1.5; P < 0.05 for both). CONCLUSIONS: After moderately painful orthopedic surgery at or distal to the elbow, 0.2% ropivacaine delivered as a continuous infusion combined with patient-controlled bolus doses via an infraclavicular perineural catheter optimizes analgesia while minimizing oral analgesic use compared with basal- or bolus-only dosing regimens. PMID- 14739818 TI - Quantifying net staffing costs due to longer-than-average surgical case durations. AB - BACKGROUND: Anesthesiology departments incur staffing costs that are not covered by revenue because the operating room (OR) time allocation and case scheduling are not done to maximize OR efficiency and because surgical durations are longer than average. The purpose of this article is to demonstrate a method to quantify net anesthesia staffing costs due to longer-than-average surgical durations and evaluate the factors that influence staffing costs. METHODS: Data collected from two anesthesiology departments in academic hospitals for 1 yr included date of surgery, time that patients entered the OR, time that patients exited the OR, surgical service, and the Current Procedural Terminology code for the primary surgical procedure. Anesthesia care performed outside the main surgical suite and services not billed with American Society of Anesthesiologists units were excluded. National average surgical durations were determined from the Current Procedural Terminology code from the Centers for Medicare and Medicaid Services' database. Actual surgical durations were then used to determine staffing solutions to maximize OR efficiency; national average surgical durations were then used to determine a second solution. The difference in staffing costs between these two staffing solutions represented the staffing costs attributable to longer surgical durations. Costs were converted to dollar amounts using compensation values reported in a national compensation survey. The differences in revenue were determined by applying conversion factors to the differences in surgical durations. The annual net cost attributable to longer surgical durations equaled the staffing costs minus the revenue produced by longer durations. Net staffing costs were estimated for two hospitals using median staffing compensation and median payer mix. Net staffing costs were then recalculated by varying the parameters (conversion factors, limits on differences between actual and average surgical duration, levels of compensation, surgical service size of OR allocation). RESULTS: Using the median compensation of staff and an average conversion factor, the net annual staffing costs attributable to longer surgical durations were $672,100 for the first hospital. However, if staff members were highly compensated and the payer mix was unfavorable, the net staffing costs were $1,688,000. Reducing the difference between actual and average duration resulted in lower staffing costs. Net staffing costs were less in a second hospital studied that had many low-volume surgical services. CONCLUSIONS: Longer-than average surgical durations can increase net staffing costs for anesthesiology groups. The increase is dependent on factors such as staffing compensation and payer mix. PMID- 14739819 TI - Chronobiology and anesthesia. PMID- 14739820 TI - Consciousness unbound: toward a paradigm of general anesthesia. PMID- 14739821 TI - Catecholamine-induced changes in the splanchnic circulation affecting systemic hemodynamics. PMID- 14739822 TI - Cerebral sinus thrombosis in a trauma patient after recombinant activated factor VII infusion. PMID- 14739823 TI - Argatroban as anticoagulant in cardiopulmonary bypass in an infant and attempted reversal with recombinant activated factor VII. PMID- 14739824 TI - Epidural granuloma and intracranial hypotension resulting from cervical epidural steroid injection. PMID- 14739825 TI - Methemoglobinemia after a blast injury. PMID- 14739826 TI - Effect of morphine on small bowel propulsion after intestinal ischemia. PMID- 14739827 TI - Obligate acceleromyography and pharmacologic reversal of all neuromuscular blocking agents: really, and where is the clinical outcome? PMID- 14739828 TI - Evidence-based practice and neuromuscular monitoring. PMID- 14739829 TI - Neuromuscular monitoring advancement. PMID- 14739830 TI - Alpha-stat induced alkalosis: cause of neuronal apoptosis after deep hypothermic perfusion. PMID- 14739832 TI - Genetic markers: not yet ready for prime time. PMID- 14739831 TI - Residual neuromuscular blockade: importance of upper airway integrity. PMID- 14739833 TI - Base excess and strong ion difference: clinical limitations related to inaccuracy. PMID- 14739834 TI - Alternative formula for Laryngeal Mask Airway trade mark size selection. PMID- 14739835 TI - Delayed postoperative arousal following remifentanil-based anesthesia in a myasthenic patient undergoing thymectomy. PMID- 14739836 TI - [Radiological geriatrics: an emergent specialty and a future specialty]. PMID- 14739837 TI - [The elderly and dementia: clinical approach]. AB - When you take care of elderly, you need to evaluate cognitive functions due to the frequency of dementia. However, before this evaluation, you must be aware of difficulties that you will encounter. You have to determine general characteristics of elderly patients and use geriatric reasoning in order to list pathologies that can interfere with cognitive functions. Thereafter, you need to precise frontiers between normal cognitive functions and dementia. When dementia is confirmed, CT scan or MRI appears very useful to exclude a cause of reversible dementia or to precise the nature of dementia. CT scan or MRI are useful during dementia to define the etiology of dementia or acute deterioration during evolution. A close collaboration between radiologist and clinician is mandatory to avoid diagnostic errors. PMID- 14739838 TI - [Dementia: role of imaging]. AB - This article summarizes the role of imaging in the evaluation of cognitive disorders of the elderly. The primary goal of imaging is to exclude potentially reversible dementia due to intracranial lesions such as tumor, subdural hematoma or normal pressure hydrocephalus. The value of neuroimaging in the diagnosis of degenerative disorders is then discussed: Alzheimer's disease, frontotemporal dementias, dementia with Lewy bodies, dementia associated with parkinsonian syndromes, vascular dementias and mild cognitive impairment. PMID- 14739839 TI - [Chest diseases in elderly: role of imaging and clinical features]. AB - Chest x-ray is the most useful examination in the elderly as community acquired pneumonia is very frequent in this high risk population. Technical details have to be considered due to the clinical status of elderly patients. Raising arms above the head may help in the radiographic diagnosis of pneumonia. Guidelines for good radiological practice indicate that a chest x-ray should be done in patients with acute respiratory illness and dementia. Clinical and biological findings of pneumonia may be misleading in the elderly and treatment should be instituted rapidly to avoid complications. Lung cancer of elderly patients has to be investigated and treated without consideration for age. Surgery should be done whenever the tumor is removable by lobectomy. Chemotherapy may be safe even in patients older than 80 with an acceptable quality of life. PMID- 14739840 TI - [Gastrointestinal tract in the elderly]. AB - Patient's age no longer is a major factor when discussing the diagnosis and treatment of hepatobiliary and gastrointestinal diseases. However, discussing with the physician in charge of the patient is the only adequate way to propose the quickest, least invasive and most comfortable imaging modality that will give a proper answer to the clinical problem. Cross sectional imaging, especially ultrasound, CT and in some cases MRI, plays an important role in the diagnosis of these diseases. PMID- 14739841 TI - [Micturition disorders in elderly patients]. AB - After clinical evaluation, imaging is extremely important in determining the type of treatment and approach of micturition disorders in elderly. In this article, the advantages and limitations of each of the relevant techniques are reviewed and highlighting, with correlation to clinical evaluation. Sonography has become the most widely used technique to study bladder dysfunction. CT and MRI have specific indications. PMID- 14739842 TI - [The lower limb of elderly patients (venous and arterial diseases)]. AB - Elderly patients frequently suffer from vascular pathologies of the leg. In these more fragile patients, diagnostic work-up must cause as little trauma as possible. The clinical status and morphological appearance of the vessels make vascular investigations sometimes difficult to perform in the elderly. With arterial pathology, vascular imaging relies on numerous non invasive techniques (Duplex scan and vascular imaging techniques by reconstruction: helical CT and magnetic resonance angiography). The clinical exam, the evolution stage of arterial disease and these non invasive investigations must allow to select the patients that should benefit from an arteriography, more or less associated to revascularization. With thrombo-embolic pathology, venography has now been replaced by venous duplex scan of the lower limbs irrespective of the underlying thrombotic etiology. PMID- 14739843 TI - [Practical approach to usual rheumatologic and traumatic diseases in elderly patients]. AB - Rheumatologic and traumatic pathology is particularly important in elderly patients because it may compromise autonomy. The severity of the loss of autonomy is variable and it may be temporary or permanent. Therefore rapid and suitable care is necessary, old age in itself being an emergency factor. The purpose of this article is to develop an approach that based on presenting symptoms such as pain, mild or severe disability, or a modification of behavior in an elderly patient will enable:--to correctly evaluate the clinical findings which will guide the imaging--to not exclude a traumatic origin even if it doesn't seem obvious--to adapt the radiological examinations depending on the therapeutic possibilities--to demonstrate that plain radiographs are usually sufficient--to not deny access to CT or MRI systematically on the basis of old age. Knowledge of specific geriatric issues and correlation with imaging findings will usually improve the quality and quantity of life of elderly patients. PMID- 14739844 TI - [Current problems in geriatrics and radiology. Teleradiology: organization and evaluation]. AB - The realization of radiological examinations in the elderly population must be adjusted to the needs of patients that usually have health problems and very little patience. Its organization must be thought as a whole: appointment, reception of an awaited patient, human accompaniment allowing good cooperation, nursing support, and prompt availability of reports. Adapted equipment facilitates the examination: stretchers to raise and carry invalid patients, digital radiography. Structural linking of radiology and geriatric units with a departmental support avoids the isolation of medical and ancillary staff, reinforce the bonds with geriatricians and facilitates the access to CT before any contrast-enhanced (barium, iodinated) examination. Teleradiology is a complementary tool of making the medical support available and reinforce the interactivity with geriatricians. The simplicity of the questions in current geriatrics makes it possible to use teleradiology in daily practice. PMID- 14739845 TI - Lymphangiogenesis is pivotal to the trials of a successful cancer metastasis. PMID- 14739846 TI - Effect of surgery for chronic pancreatitis on pancreatic function: pancreatico jejunostomy and duodenum-preserving resection of the head of the pancreas. AB - BACKGROUND: Resection and drainage procedures are performed for chronic pancreatitis. After resection, pancreatic function deteriorates; however, little is known about the effect of drainage procedures. METHODS: Pancreatic function was evaluated prospectively before and after surgery in 27 patients with duodenum preserving resection of the head of the pancreas (DPRHP), and in 12 patients with pancreatico-jejunostomy (P-JS); 18 patients with chronic pancreatitis served as controls. Results of the 2 groups were not compared because of differences in patient characteristics and indications for surgery. Endpoints were exocrine function (fecal fat excretion, urinary PABA recovery), endocrine function (oral glucose tolerance test, serum C-peptide concentrations), and pancreatic polypeptide secretion. RESULTS: Groups were not different with respect to age and duration of symptoms. Median urinary PABA recovery was not altered significantly after surgery: DPRHP, from 40% to 31%; P-JS, from 52% to 44%; and controls, from 43% to 48%. Median fecal fat also did not change significantly: DPRHP, from 6 to 12 g/24 h; P-JS, from 9 to 5 g/24 h; and controls, from 6 to 7 g/24 h. Although the integrated blood glucose value did not change after DPRHP, the integrated serum C-peptide value decreased after DPRHP (P<.02). After P-JS, the integrated blood glucose value decreased (P<.02), but there was no change in integrated serum C-peptide secretion. Neither integrated blood glucose nor C peptide values were affected in controls. Insulin dependency increased (22% to 33%) after DPRHP. Pancreatic polypeptide secretion decreased only after DPRHP (P=.003). CONCLUSIONS: Surgery for chronic pancreatitis does not influence exocrine pancreatic function after either a drainage (P-JS) or a resection procedure (DPRHP). Clinical endocrine function is not affected after DPRHP but improves after P-JS. PMID- 14739847 TI - Effects of heparin in experimental models of acute pancreatitis and post-ERCP pancreatitis. AB - BACKGROUND: Acute pancreatitis (AP) is a complication of diagnostic or therapeutic endoscopic retrograde cholangiopancreatography (ERCP). In a recent clinical trial, a decreased rate of post-ERCP pancreatitis was shown after prophylactic heparin treatment. The aim of this study was to evaluate the effects of prophylactic heparin application in various experimental models of AP and pancreatic duct obstruction and to assess the underlying mechanisms. METHODS: In various experimental models, pancreatic injury of graded severity was induced in Wistar rats: (1) mild pancreatitis by IV cerulein infusion over 6 hours; (2) severe pancreatitis by infusion of glycodeoxycholic acid into the pancreatic duct plus IV cerulein application over 6 hours. The clinical ERCP situation was imitated in groups (3) obstruction of the pancreatic duct and (4) infusion of contrast medium into the pancreatic duct plus obstruction. In every group the animals received either no heparin (n=six per group) or continuous IV heparin (n=six per group) starting before pancreatic injury. Histologic changes, amylase, and lipase in plasma were evaluated 12 hours after induction of pancreatic injury. Additional animals were treated to investigate pancreatic microcirculation by intravital microscopy (n=six per group). RESULTS: In groups 1, 3, and 4 (mild AP/duct obstruction/duct obstruction plus contrast medium), IV heparin-treated animals showed reduced edema, inflammation, and peak amylase values compared with the corresponding non-heparin-treated animals (P<.05). Moreover, mean erythrocyte velocity was significantly higher and leukocyte endothelium interaction was reduced in these groups after prophylactic administration of heparin. In contrast, group 2 (severe AP) did not show any difference between control animals and animals that received heparin as assessed by histology and intravital microscopy. CONCLUSIONS: Prophylactic systemic application of heparin provides a protective effect in mild AP and in experimental post-ERCP pancreatitis. The mechanism of the protective effects of heparin seems to be the reduction of leukocyte-endothelium interaction and the normalization of pancreatic microcirculation. PMID- 14739848 TI - A novel classification system for patients with PTC: addition of the new variables of large (3 cm or greater) nodal metastases and reclassification during the follow-up period. AB - BACKGROUND: Several factors have been proven to be useful for classifying patients with papillary thyroid carcinoma (PTC) into either low- or high-risk groups. However, the relative importance of prognostic factors, including lymph nodal metastasis, remains unclear. METHODS: A total of 604 patients who underwent initial surgery for PTC (diameter of tumor>1 cm) were analyzed. The mean duration of follow-up was 10.7 years. RESULTS: By multivariate analysis for disease specific survival, distant metastasis was the only significant risk factor (risk ratio=65.1) for younger patients (age<50). For older patients (age> or =50), distant metastasis (risk ratio=6.7), extrathyroidal invasion (risk ratio=2.4), and large nodal metastasis (> or =3 cm; risk ratio=5.3) had relative importance. From the results, younger patients with distant metastasis and older patients with any of the 3 factors were defined as at high risk, whereas the other patients were defined as at low risk. Overall, 106 patients at high risk (18%) and 498 patients at low risk (83%) had 10-year survival rates of 69% and 99%, respectively. Only 3 patients of the low-risk group died from the disease. Among postoperative factors, recurrence within 3 years after initial surgery was the most important risk factor for cancer death. Of the high-risk group, patients with a disease-free interval of >3 years showed an excellent outcome (96% of a 10 year survival rate), similar to patients in the low-risk group. CONCLUSIONS: A novel classification system, in which large nodal metastases and postoperative reclassification were added, was devised. This was useful for choosing proper therapeutic strategies, offering rational information, and determining adequate postoperative follow-up schemes for individual patients with PTC. PMID- 14739849 TI - Comparison of parathyroid hormone levels from the intact and whole parathyroid hormone assays after parathyroidectomy for primary and secondary hyperparathyroidism. AB - BACKGROUND: Most commercial intact parathyroid hormone (intact PTH) assays cross react with non-(1-84) PTH (likely 7-84 PTH). Using a whole-molecule PTH (whole PTH) assay that specifically measured only 1-84 PTH, we compared the kinetics of whole PTH and intact PTH after parathyroidectomy in patients with primary hyperparathyroidism (HPT) and secondary HPT. METHODS: This study comprised 74 patients with primary HPT caused by a single adenoma and 18 patients with secondary HPT who underwent parathyroidectomy. Blood samples were drawn after anesthesia, just before excision of a single adenoma in primary HPT, and just before excision of the last parathyroid gland in secondary HPT, and at 5, 10, and 15 minutes after excision. The 7-84 PTH level was calculated by subtracting the whole PTH value from the intact PTH value. RESULTS: There was a difference between the percentage of 7-84 PTH/intact PTH in plasma samples from patients with primary HPT and secondary HPT (28%+/-12% vs 35%+/-9%; P<.05). Plasma whole PTH decreased more rapidly than intact PTH after parathyroidectomy in patients in both the primary HPT (P<.0001) and secondary HPT groups (P<.0001). Decline of intact PTH was slower in patients with secondary HPT than in patients with primary HPT; however, there was no significant difference in the decline of whole PTH between the 2 groups. CONCLUSIONS: The quick intact PTH assay is not used frequently during surgery in patients with secondary HPT; however, our results suggest that a quick whole PTH assay may be a more useful adjunct to parathyroidectomy in both secondary HPT and primary HPT. PMID- 14739850 TI - CT-MIBI image fusion: a new preoperative localization technique for primary, recurrent, and persistent hyperparathyroidism. AB - BACKGROUND: Successful minimally invasive or imaging-guided operations in patients with primary, recurrent, and persistent hyperparathyroidism are based on the reliability of preoperative parathyroid localization studies. The CT-MIBI image fusion promises a higher diagnostic accuracy than current imaging procedures. The aim of our study was to assess its reliability in correctly detecting enlarged parathyroid glands. METHODS: In a prospective study 24 consecutive patients underwent CT-MIBI image fusion as preoperative parathyroid localization procedure. The results of technetium 99m sestamibi single photon emission computed tomography (MIBI-SPECT) alone, today the standard method in parathyroid imaging, and CT-MIBI image fusion were analyzed by a blinded reviewer, and the imaging results were compared with the intraoperative findings. RESULTS: For CT-MIBI image fusion a sensitivity of 93% and a specificity of 100% in correctly detecting the position of enlarged parathyroid glands was calculated and compared with a sensitivity of MIBI-SPECT of 31% and a specificity of 87% (P<.001). This new imaging technique enabled us to successfully treat 22 of our patients (92%) with imaging-guided surgery. Twenty (83%) underwent unilateral or minimally invasive operations. CONCLUSIONS: CT-MIBI image fusion appears to be superior to MIBI-SPECT in preoperative parathyroid imaging. CT-MIBI image fusion can be performed on existing CT- and MIBI-SPECT units. We recommend this method for preoperative localization in patients with primary, recurrent and persistent hyperparathyroidism. PMID- 14739851 TI - Protrusion of prosthetic meshes in repair of inguinal hernias. AB - BACKGROUND: Although techniques with prosthetic meshes have improved results of inguinal hernia repair, the problem of recurrence remains. In addition to patient and surgeon-related causes, protrusion of prosthetic mesh through the hernia defect can be considered as a factor of recurrence. METHODS: To simulate inguinal hernia, porcine tissue with standardized defects (3, 4, and 5 cm) was used. Nine different meshes (9 cm diameter) were positioned on the defects. In a pressure chamber the depth of protrusion through the defect was assessed. Both sides of each mesh were measured. Protrusion at a pressure representing Valsalva pressure was used for statistical analysis. RESULTS: Protrusion, including the incidence of collapse, increased with larger defect sizes. Significant protrusion differences were found between different meshes and occasionally between the 2 sides of the same mesh. CONCLUSIONS: The differences between the effects of the various meshes emphasize the importance of material characteristics when developing new surgical techniques. The 3 meshes showing least protrusion are considered especially suitable when repairing large defects or when preferring a small mesh when repairing inguinal hernias with a preperitoneal sublay approach. PMID- 14739852 TI - Palliation of anastomotic obstructions in recurrent gastric carcinoma with the use of covered metallic stents: clinical results in 25 patients. AB - BACKGROUND: The purpose of this study was to investigate the technical feasibility and the clinical effectiveness of the placement of covered self expandable metallic stents for the treatment of anastomotic obstructions in recurrent gastric carcinoma. METHODS: With fluoroscopic guidance, covered stents were placed in 25 patients with recurrent gastric carcinoma for the palliation of obstructions at anastomotic sites (14 gastrojejunostomy, 11 esophagojejunostomy). All patients had severe nausea and recurrent vomiting before the stent placement. RESULTS: Stent placement was technically successful in 24 patients (96%). After stent placement, symptoms improved in all 24 patients. During the follow-up of 2 to 65 weeks (mean, 13.7 weeks), stent migration occurred in 1 patient 16 days after the procedure. He needed percutaneous catheter drainage because of an abscess, which was followed by esophagojejunostomy site rupture during a second stent trial. Stricture recurred in 4 patients because of tumor overgrowth 10 to 55 weeks after the procedure; all patients underwent coaxial placement of a second stent and had good oral intake. CONCLUSIONS: The placement of covered expandable metallic stents seems to be both technically feasible and an effective means for the palliation of anastomotic obstructions in recurrent gastric carcinoma. This procedure can be considered to be the primary choice for the palliation in those patients. PMID- 14739853 TI - Modification of xenoantigens on porcine erythrocytes for xenotransfusion. AB - BACKGROUND: Problems of supply and disease transmission with blood transfusion may be controlled by the use of an isolated animal donor pool. However, porcine erythrocytes (PRBCs) usually are destroyed rapidly by preformed antibodies in human serum. We examined the impact on PRBC antigenicity by the removal of cell membrane alpha-gal(1-3)beta-galGlcNac epitopes (called alpha-gal) and chemical masking of other xenoantigens. METHODS: From porcine "low expressors" of alpha gal, PRBCs were subjected to (1) enzymatic removal of membrane alpha-gal with alpha-galactosidase, (2) covalent attachment of cyanuric acid-linked methoxypolyethylene glycol, or (3) both processes. PRBC integrity was assessed by light microscopy, scanning electron microscopy, osmotic fragility, and determination of oximetric p50. The effects of treatment were measured by hemagglutination, complement fixation, flow cytometric assay of immunoglobulin G/M binding, and clinical cross-match testing to human sera. RESULTS: Cyanuric acid-linked methoxypolyethylene glycol reduced hemagglutination titers moderately, although alpha-galactosidase treatment reduced hemagglutination titers to levels similar to negative controls. The combination of the treatments was most effective, by the reduction of binding of human immunoglobulin M by 61% compared with controls. RBC morphologic condition, stability, and p50 values were maintained. Clinically used cross-match tests between PRBCs and human sera demonstrated increased compatibility. CONCLUSIONS: These data suggest that strategies to remove or mask xenoantigens on PRBCs reduce antigenicity sufficiently to allow in vitro cross-match compatibility to human sera. PMID- 14739854 TI - Intra-abdominal sepsis attenuates local inflammation-mediated increases in microvascular permeability at remote sites in mice in vivo. AB - BACKGROUND: Given that leukocyte delivery to remote sites is diminished in states of systemic inflammation, such as sepsis, and activated leukocytes may be responsible for endothelial injury leading to vascular leakage, we hypothesized that intra-abdominal sepsis would diminish microvascular leakage at remote sites by altering leukocyte-endothelial interactions. METHODS: Using a murine intravital microscopy model, we examined leukocyte-endothelial interactions and vascular leakage at a peripheral site in the presence of local and/or systemic inflammation. Forty mice were randomized to 1 of 4 study groups: local infection (orchitis), systemic infection (intra-abdominal sepsis by cecal ligation and puncture), local and systemic infection, and control. Postcapillary venules of the cremaster muscle were examined by bright light and fluorescence intravital microscopy. Microvascular leakage was determined after intravenous administration of fluorescent albumin. RESULTS: Systemic infection attenuated the increases in both leukocyte adherence and local infection-induced microvascular permeability. Neutrophil cell-surface expression of L-selectin, as determined by flow cytometry, diminished with both local and systemic infection, whereas expression of CD11b increased with systemic, but not local, infection. CONCLUSIONS: These data suggest that systemic (intra-abdominal) sepsis diminishes local inflammation mediated vascular leakage by attenuating leukocyte adherence. PMID- 14739855 TI - Insulin decreases hepatic acute phase protein levels in severely burned children. AB - BACKGROUND: Severe burn induces the hepatic acute phase response. In this study, we wondered whether continuous insulin treatment decreases acute phase protein levels in the severely burned. METHODS: Eighteen children aged 2 to 17 years with burns >40% of total body surface area were randomized to receive either insulin (n=9) or no treatment (n=9) within 72 hours after injury until the wounds were 95% healed. Insulin was given at a continuous rate of > or =1.5 microU/kg/min to maintain euglycemia (serum glucose 100-140 microg/dL). Plasma was examined at days 7, 14, 21, and 28 for acute phase protein levels including C-reactive protein, C3 complement, alpha1-acid glycoprotein, haptoglobin, alpha2 macroglobulin, prealbumin, transferrin, and retinol-binding protein. Statistical analysis was by ANOVA and t test. RESULTS: With insulin treatment, alpha1-acid glycoprotein, C3 complement, alpha2-macroglobulin, and haptoglobin levels decreased (P<.05) after a severe burn compared with control, especially at days 21 and 28. Additionally, the hepatic constitutive proteins (prealbumin, transferrin, and retinol-binding protein) were lower in the insulin-treatment group than those of the control group at day 21 (P<.05). CONCLUSIONS: Continuous insulin treatment decreases acute phase protein levels after a severe burn. The results suggest insulin downregulation of the hepatic acute phase response to injury. PMID- 14739856 TI - Ischemic preconditioning and methylprednisolone both equally reduce hepatic ischemia/reperfusion injury. AB - BACKGROUND: Ischemic preconditioning (I/P) and methylprednisolone (MP) have been suggested to protect against ischemia-reperfusion (IR) injury, which results in an increased tolerance against organ hypoxia. METHODS: Before 45 minutes of hepatic ischemia, male Wistar rats were pretreated with either I/P (5/30 minutes) or MP (30 mg/kg BW). The degree of IR injury and the postischemic inflammatory (leukocyte infiltration, myeloperoxidase, intercellular adhesion molecule-1) and apoptotic (TUNEL, caspase 3, cytochrome C) activity was measured in both groups and compared with non-pretreated (ischemic) animals. RESULTS: Histology and enzyme release revealed that I/P and MP treatment provided significant protection as compared with ischemic controls. TUNEL-positive cells, as well as caspase 3 and cytochrome C expression, were clearly reduced in hepatic tissue of MP-treated animals and partially reduced in I/P-treated animals when compared with ischemic animals. The inflammatory response was considerably reduced in MP- and I/P treated animals, especially in the early period after ischemia. NF-kappaB/Rel binding activity was increased after I/P and decreased in MP-treated animals, whereas ischemic controls showed a constant binding activity. CONCLUSIONS: MP (probably by downregulation of NF-kappaB-binding activity) and I/P attenuated the postischemic apoptotic and inflammatory response. Both treatments equally reduced IR-related hepatocellular damage, and, thus, may also be applied equally in surgery involving warm organ hypoxia. PMID- 14739857 TI - Taurodeoxycholate increases intestinal epithelial cell proliferation through c myc expression. AB - BACKGROUND: Bile salts have been shown to modulate gastrointestinal epithelial restitution, differentiation, and other functions. Prior studies have shown that the bile salt taurodeoxycholate increases cell migration after injury. The purpose of this experiment was to determine the effect that taurodeoxycholate has on intestinal epithelial cell growth, c-myc expression and function. METHODS: IEC 6 or Caco-2 cells were treated with varying concentrations of taurodeoxycholate (.05 to 1 mmol/L) and proliferation determined. Apoptosis was measured by use of DNA fragmentation assay and nuclear staining. Cell phase was determined with propidium iodide flow cytometry. C-myc expression was determined by Northern and Western blot analysis, and c-myc function was inhibited by specific c-myc antisense. RESULTS: There was no change in cell structure. Apoptosis was not induced. Six days after exposure to taurodeoxycholate, IEC-6 cell proliferation was significantly increased. Flow cytometry showed a significant increase in S phase concentration and a significant decrease in G1-phase concentration of the cell cycle. Taurodeoxycholate also increased c-myc protein and mRNA expression, and inhibition of c-myc function prevented taurodeoxycholate-induced cell proliferation. CONCLUSIONS: Exposure to physiological concentrations of the bile salt taurodeoxycholate increases intestinal epithelial cell proliferation. This effect is at least partially mediated through a c-myc-dependent mechanism. Bile salts can have a beneficial effect on the intestinal mucosa. PMID- 14739858 TI - Tapering duodenoplasty and Roux-en-Y duodenojejunostomy in the management of adult megaduodenum. PMID- 14739859 TI - Wilkie's syndrome. PMID- 14739860 TI - Retroperitoneal benign cystic peritoneal mesothelioma. PMID- 14739862 TI - Protected time. PMID- 14739863 TI - Physician bias about participation in clinical trials? PMID- 14739864 TI - Prognostic factors in papillary thyroid carcinoma and implications of large nodal metastasis. PMID- 14739869 TI - Biomechanical and neurophysiological responses to spinal manipulation in patients with lumbar radiculopathy. AB - OBJECTIVE: The purpose of this study was to quantify in vivo vertebral motions and neurophysiological responses during spinal manipulation. METHODS: Nine patients undergoing lumbar decompression surgery participated in this study. Spinal manipulative thrusts (SMTs) ( approximately 5 ms; 30 N [Sham], 88 N, 117 N, and 150 N [max]) were administered to lumbar spine facet joints (FJs) and spinous processes (SPs) adjacent to an intraosseous pin with an attached triaxial accelerometer and bipolar electrodes cradled around the S1 spinal nerve roots. Peak baseline amplitude compound action potential (CAP) response and peak-peak amplitude axial (AX), posterior-anterior (PA), and medial-lateral (ML) acceleration time and displacement time responses were computed for each SMT. Within-subject statistical analyses of the effects of contact point and force magnitude on vertebral displacements and CAP responses were performed. RESULTS: SMTs (>/= 88 N) resulted in significantly greater peak-to-peak ML, PA, and AX vertebral displacements compared with sham thrusts (P <.002). SMTs delivered to the FJs resulted in approximately 3-fold greater ML motions compared with SPs (P <.001). SMTs over the SPs resulted in significantly greater AX displacements compared with SMTs applied to the FJs (P <.05). Seventy-five percent of SMTs resulted in positive CAP responses with a mean latency of 12.0 ms. Collectively, the magnitude of the CAP responses was significantly greater for max setting SMTs compared with sham (P <.01). CONCLUSIONS: Impulsive SMTs in human subjects were found to stimulate spinal nerve root responses that were temporally related to the onset of vertebral motion. Further work, including examination of the frequency and force duration dependency of SMT, is necessary to elucidate the clinical relevance of enhanced or absent CAP responses in patients. PMID- 14739870 TI - Adverse reactions to chiropractic treatment and their effects on satisfaction and clinical outcomes among patients enrolled in the UCLA Neck Pain Study. AB - BACKGROUND: Minor side effects associated with chiropractic are common. However, little is known about their predictors or the effects of reactions on satisfaction and clinical outcomes. OBJECTIVE: The objectives of this study are to compare the relative effects of cervical spine manipulation and mobilization on adverse reactions and to estimate the effects of adverse reactions on satisfaction and clinical outcomes among patients with neck pain. METHODS: Neck pain patients were randomized to receive cervical spine manipulation or mobilization. At 2 weeks, subjects were queried about possible treatment-related adverse reactions and followed for 6 months with assessments for pain and disability at 2, 6, 13, and 26 weeks. Numerical rating scales and the Neck Disability Index were used to measure pain and disability. Perceived improvement and satisfaction with care were assessed at 4 weeks. RESULTS: Of 960 eligible patients, 336 enrolled and 280 responded to the adverse event questionnaire. Thirty percent of respondents reported at least 1 adverse symptom, most commonly increased pain and headache. Patients randomized to manipulation were more likely than those randomized to mobilization to report an adverse reaction (adjusted odds ratio = 1.44, 95% confidence interval = 0.85, 2.43). Subjects reporting adverse reactions were less satisfied with care and less likely to have clinically meaningful improvements in pain and disability. CONCLUSIONS: Adverse reactions are more likely to be reported following cervical spine manipulation than mobilization. Chiropractors may reduce iatrogenesis and increase satisfaction and perhaps clinical outcomes by mobilizing rather than manipulating their neck pain patients. PMID- 14739871 TI - Assessing the clinical significance of change scores recorded on subjective outcome measures. AB - BACKGROUND: To date, clinical trials have relied almost exclusively on the statistical significance of changes in scores from outcome measures in interpreting the effectiveness of treatment interventions. It is becoming increasingly important, however, to determine the clinical rather than statistical significance of these change scores. OBJECTIVE: To determine cutoff values for change scores that distinguish patients who have clinically improved from those who have not. METHOD: Data were obtained from 165 back and 100 neck patients undergoing chiropractic treatment. Patients completed the Bournemouth Questionnaire (BQ) before treatment and the BQ and Patient's Global Impression of Change (PGIC) scale after treatment. Three statistical methods were applied to individual change scores on the BQ. These were (1) the Reliable Change Index (RCI); (2) the effect size (ES); and (3) the raw and percentage change scores. The PGIC scale was used as the "gold standard" of clinically significant change. RESULTS: The RCI, using the cutoff value of >1.96, appropriately identified clinical improvement in back patients but not in neck patients. An individual ES of approximately 0.5 had the highest sensitivity and specificity in distinguishing back and neck patients who had undergone clinically significant improvement from those who had not. In terms of raw score changes, percentage BQ change scores [(raw change score/baseline score) x 100] of 47% and 34% were identified as having the highest sensitivity and specificity in distinguishing clinically significant improvement from nonimprovement in back and neck patients, respectively. CONCLUSION: This study provides a methodological framework for identifying clinically significant change in patients. This approach has important implications in providing clinically relevant information about the effect of a treatment intervention in an individual patient. PMID- 14739872 TI - Guidance hypothesis with verbal feedback in learning a palpation skill. AB - OBJECTIVE: To investigate whether a force application of a novel motor skill was simple or complex and to determine the appropriate level of feedback during training. The objective was to determine the effects of various amounts of knowledge of results (KR) on learning a novel skill that is frequently taught in chiropractic for the assessment of vertebral motor unit dysfunction. METHODS: Thirty-five active subjects were taught the novel skill of spring testing to a particular force range through 9 or 10 teaching sessions over a 2-week period (a school holiday limited some to only 9 sessions). To determine the particular force range, an initial phase of the study involved a licensed and practicing clinician performing motion palpation spring testing of the thoracic spine of a prone subject. The data from a total of 47 pushes throughout the thoracic spine were recorded, and the mean force and standard deviation were calculated. The second phase of the study used the obtained mean and standard deviation for teaching the force of prone thoracic spine motion palpation to 35 active subjects by administering spring testing to 35 passive subjects. The active subjects were randomly divided into 4 groups, with each receiving a varying amount of verbal feedback to move toward the target force of their learned skill. Each passive subject was laid prone on an instrumented adjusting table. Group 1 received the least amount of feedback while learning the novel skill. Groups 2 and 3 received more frequent, intermittent feedback while learning the skill, and group 4 received constant, frequent feedback during each of the 10 teaching sessions. All subjects returned within 2 weeks for 2 retention trials to determine the efficacy of the learned skill. RESULTS: The mean force determined was 143 N with a standard deviation of 14 N. Each subject was taught spring testing within this target range. The 2 final retention trials showed group 3 to have demonstrated the most targeted retention of the learned motor skill. These subjects exhibited the closest force range to the target for the motor skill learned and fewer changes in standard deviation compared with their acquisition trials and thus the highest retention. Group 4, receiving 100% feedback, demonstrated the most accurate spring testing during the acquisition trials, but the subjects did not effectively demonstrate when compared on acquisition and retention trials. CONCLUSION: The results of this study show data to be consistent with the guidance hypothesis in learning a novel motor skill. The constant KR feedback is beneficial for learning when used to reduce error during practice but detrimental when relied upon for retention and learning. These data suggest the necessity of using motor skill development learning theory in the teaching of chiropractic. PMID- 14739873 TI - Response rates for surveys of chiropractors. AB - BACKGROUND: Survey response rates may vary by type of practitioner studied and may have declined over time. Response rates for surveys of complementary practitioners have not been studied. OBJECTIVE: To describe the response rates in published surveys of chiropractors and explore for secular trends in response rates and for methodologic and geographic correlates of response rates. METHODS: Secondary analysis of data extracted from published English language reports of surveys of chiropractors. Response rates were calculated as the total number of persons from whom a questionnaire was returned divided by the total number of persons who were sent a questionnaire. RESULTS: Sixty-two surveys represented by 79 articles published in the interval 1980 to 2000 met inclusion criteria for analysis. We were able to calculate a response rate for 46 postal surveys. The mean response rate was 52.7%. There was no significant association between geographic setting and response rate, and there was no evidence of secular trend in response rates. None of the studies employed incentives. The strongest predictor of response rate was number of contacts with the target population. CONCLUSION: Response rates for surveys of chiropractors are similar to those observed for surveys of medical doctors. The key to obtaining high response rates is the use of evidence-based methods in design and conduct of the surveys. PMID- 14739874 TI - The forces applied by female and male chiropractors during thoracic spinal manipulation. AB - BACKGROUND: On average, women weigh less, have a smaller frame, and are less muscular than men. Since the peak thrust force applied during spinal manipulative treatments can be quite high and must be reached in a very short period of time, one might question the physical ability of women to generate such high forces. OBJECTIVE: To study the forces generated by male and female chiropractors as they deliver spinal manipulation to the thoracic spine. METHODS: Fourteen male and 14 female experience-matched chiropractors participated in this study. They each manipulated 1 of 9 asymptomatic male adult subjects of similar height and weight. The clinicians were asked to manipulate a transverse process in the vicinity of T4 and T9. Any technique could be used as long as the treatment thrust was in a posterior to anterior direction and the hand contact fit onto the sensor pad (area = 100 cm2). RESULTS: There were no significant differences (P<.05) between male and female chiropractors for any measurements in the upper thoracic area. For the lower thoracic manipulations, the preload forces for the male chiropractors were significantly greater (P<.05) than those for the female chiropractors. The remaining variables were the same between the 2 groups. CONCLUSIONS: Female chiropractors produce, from a mechanical point of view, similar manual treatments as their male colleagues. PMID- 14739876 TI - Biomechanical assessments of lumbar spinal function. How low back pain sufferers differ from normals. Implications for outcome measures research. Part I: kinematic assessments of lumbar function. AB - OBJECTIVE: To review new and advanced biomechanical assessment techniques for the lumbar spine and illustrate the differences in lumbar function in patients with low back pain and asymtomatic subjects. DATA SOURCES: The biomedical literature was searched for research and reviews on spinal kinematic differences between low back pain subjects and healthy controls. A data search for articles indexed on MEDLINE until April 2002 was performed. RESULTS: Kinematic measurements of lumbar function were categorized into 3 areas where low back patients may differ from normals: (1) end range of motion during simple movements; (2) higher order kinematics (displacement, velocity, and acceleration) during complex movement tasks; and (3) spinal proprioception. The assessment of higher order kinematics during complex movement tasks is the most highly researched and the most successful in describing differences between the populations. The use of simple end range of motion appears questionable, while assessing spinal proprioception is the least researched, yet shows potential in highlighting differences between low back sufferers and asymptomatics. CONCLUSION: Current kinematic biomechanical assessment techniques are capable of identifying functional differences between low back pain populations and controls. The use and validity of the majority of these techniques as outcome measures are currently unknown, yet may be valuable in generating functional diagnoses, evaluating the mechanisms of current therapies, and prescribing specific rehabilitation programs. PMID- 14739877 TI - Routine screening for abuse: opening Pandora's box? AB - OBJECTIVE: To review the case of a patient who had been physically abused by a previous partner and to discuss the importance of routine screening for abuse. CLINICAL FEATURES: A 33-year-old female had neck pain, right shoulder pain, and low back pain. During the history taking, she disclosed that she had been in an abusive relationship, and her current complaints were thought to be a direct result of one of the violent episodes she suffered. Radiographs of her neck and shoulder were obtained to rule out a traumatic etiology. In the absence of any gross abnormalities, her symptoms were found to be biomechanical in origin. INTERVENTION AND OUTCOME: Therapy included spinal manipulation and soft tissue techniques to the symptomatic areas. During the treatments, much consideration was given to the fact that she had been a victim of domestic violence and the emotional and physical impact this had on her current situation. Manipulative techniques were adapted so that the patient would not perceive them as threatening, and a considerable amount of time was spent on counseling. After a number of treatments, she reported significant improvement in her overall condition. CONCLUSIONS: For practitioners of spinal manipulative therapy, routine screening for abuse is an essential part of history taking. Awareness of a patient's experiences with domestic violence is required to make appropriate adjustments in the management plan. PMID- 14739878 TI - Internal forces sustained by the vertebral artery during spinal manipulative therapy. PMID- 14739879 TI - Internal forces by the vertebral artery during spinal manipulative therapy. PMID- 14739880 TI - User's guide to the chiropractic literature-1A: how to use an article about therapy. PMID- 14739881 TI - Several pathways in the evolution of chiropractic manipulation. PMID- 14739882 TI - Can patient reactions to the first chiropractic treatment predict early favorable treatment outcome in persistent low back pain? PMID- 14739884 TI - Evaluation of Chapman's neurolymphatic reflexes via applied kinesiology: a case report of low back pain and congenital intestinal abnormality. AB - OBJECTIVE: To describe the applied kinesiologic evaluation of Chapman's neurolymphatic (NL) reflexes in the management of a person with an unusual congenital bowel abnormality and its role in the manifestation of low back pain. The theoretical foundations of these reflexes will be elaborated on and practical applications discussed. CLINICAL FEATURES: A 29-year-old man had chronic low back pain. Radiographs of the patient's lumbar spine and pelvis were normal. Magnetic resonance imaging (MRI) demonstrated a mild protrusion of the fifth lumbar disk. Oral anti-inflammatory agents, cortisone injections, and chiropractic manipulative therapy provided little relief. Though generally in robust health, the patient was aware of a congenital intestinal abnormality diagnosed when he was a child; it was thought to be of no consequence with regard to his current back condition. INTERVENTION AND OUTCOME: The patient's history, combined with applied kinesiology examination, indicated a need to direct treatment to the large bowel. The essential diagnostic indicators were the analysis of the Chapman's neurolymphatic reflexes themselves, coupled with an evaluation of the traditional acupuncture meridians. The primary prescribed therapy was the stimulation of these reflexes by the patient at home. This intervention resulted in the resolution of the patient's musculoskeletal symptomatology, as well as improved bowel function. CONCLUSION: The rather remarkable outcome from the application of this relatively simple, yet valuable, diagnostic and therapeutic procedure represents a thought-provoking impetus for future study and clinical application. PMID- 14739885 TI - Occlusion in removable partial prosthodontics. 1972. PMID- 14739886 TI - Restoration of unfavorably positioned implants for a partially endentulous patient by using an overdenture retained with a milled bar and attachments: a clinical report. AB - The restorative dentist may encounter patients with previously placed misaligned implants that are well integrated. Often, these patients have deficient alveolar ridge contour further complicating treatment options. This clinical report describes a treatment approach for a partially edentulous patient with unfavorably positioned implants that incorporated: (1) an implant-supported milled bar with a removable, metal-acrylic resin, partial overdenture, (2) implant-supported single crowns, and (3) crowns retained by natural teeth. PMID- 14739887 TI - Use of polyethylene ribbon to create a provisional fixed partial denture after immediate implant placement: a clinical report. AB - Provisional restorations are generally necessary to restore lost function and esthetics during the implant integration period. This article describes the fabrication of an esthetic, economic, and conservative bonded provisional fixed partial denture for a patient restored with implants. A reinforcing polyethylene ribbon was used to bond the natural lateral incisors to the adjacent teeth. An acceptable esthetic outcome was obtained, and the result was satisfactory to the patient. PMID- 14739888 TI - Observation of horizontal mandibular positions in an edentulous patient using a digital gothic arch tracer: a clinical report. AB - Dentures were fabricated for a 73-year-old woman using porcelain 20-degree maxillary posterior teeth and acrylic resin flat planes in the mandibular posterior region. A digital gothic arch tracing device was used to observe the horizontal mandibular positions before insertion and to evaluate the therapeutic effect of the diagnostic dentures at 1 and 3 months after insertion. PMID- 14739889 TI - Influence of implant length and diameter on stress distribution: a finite element analysis. AB - STATEMENT OF PROBLEM: Masticatory forces acting on dental implants can result in undesirable stress in adjacent bone, which in turn can cause bone defects and the eventual failure of implants. PURPOSE: A mathematical simulation of stress distribution around implants was used to determine which length and diameter of implants would be best to dissipate stress. MATERIAL AND METHODS: Computations of stress arising in the implant bed were made with finite element analysis, using 3 dimensional computer models. The models simulated implants placed in vertical positions in the molar region of the mandible. A model simulating an implant with a diameter of 3.6 mm and lengths of 8 mm, 10 mm, 12 mm, 14 mm, 16 mm, 17 mm, and 18 mm was developed to investigate the influence of the length factor. The influence of different diameters was modeled using implants with a length of 12 mm and diameters of 2.9 mm, 3.6 mm, 4.2 mm, 5.0 mm, 5.5 mm, 6.0 mm, and 6.5 mm. The masticatory load was simulated using an average masticatory force in a natural direction, oblique to the occlusal plane. Values of von Mises equivalent stress at the implant-bone interface were computed using the finite element analysis for all variations. Values for the 3 most stressed elements of each variation were averaged and expressed in percent of values computed for reference (100%), which was the stress magnitude for the implant with a length of 12 mm and diameter of 3.6 mm. RESULTS: Maximum stress areas were located around the implant neck. The decrease in stress was the greatest (31.5%) for implants with a diameter ranging from of 3.6 mm to 4.2 mm. Further stress reduction for the 5.0 mm implant was only 16.4%. An increase in the implant length also led to a decrease in the maximum von Mises equivalent stress values; the influence of implant length, however, was not as pronounced as that of implant diameter. CONCLUSIONS: Within the limitations of this study, an increase in the implant diameter decreased the maximum von Mises equivalent stress around the implant neck more than an increase in the implant length, as a result of a more favorable distribution of the simulated masticatory forces applied in this study. PMID- 14739890 TI - Passive tactile sensibility in edentulous subjects treated with dental implants: a pilot study. AB - STATEMENT OF PROBLEM: Edentulous patients treated with implant-supported prostheses have shown increased passive tactile sensibility compared with those using conventional complete dentures. This is thought to be due to the close mechanical coupling between the implant and bone via the osseointegrated interface, yet the phenomenon has received little attention. PURPOSE: The purpose of this study was to measure passive tactile sensibility in a group of edentulous subjects treated with dental implants, and to relate the measured sensibility to a range of factors thought to be of possible relevance, namely, patient age, gender, time since implant placement, implant length, and implant separation. MATERIAL AND METHODS: Twenty edentulous subjects successfully treated with 2 or more Nobel Biocare dental implants in the anterior mandible were studied. The inclusion criteria were : (1) age of less than 50 years, (2) a period of at least 12 months since implant placement, (3) implant length of at least 10 mm and of standard diameter (excluding narrow and wide platform designs), and (4) implant separation of at least 18 mm. Using a computer-controlled custom-made device, pushing forces (2.1, 2.4, 2.7, and 3.0 N/s) were applied directly and perpendicular to the long axes of the implant abutments until the subjects felt the first sensation of pressure. The magnitude of these forces was measured with an integral transducer. The applied force had a ramped staircase pattern, and force application rates were varied between 2.1 and 3.0 N/s. Multilevel modeling was used to analyze the collected data (alpha=.05). RESULTS: The threshold values of passive tactile sensibility ranged between 3.1 and 15.7 N (mean 10.9; SD 3.9). Analysis failed to show any significant association between passive tactile sensibility and the variables studied. CONCLUSION: Within the limitations of this study, which included a small sample size, no relationship was found between passive tactile sensibility associated with long-standing implants and any of the variables studied (age, gender, time since implant placement, implant length, and implant separation). PMID- 14739891 TI - The effectiveness of auxiliary features on a tooth preparation with inadequate resistance form. AB - STATEMENT OF PROBLEM: No study has evaluated the efficacy of auxiliary tooth preparation elements for crowns with originally reduced resistance form. PURPOSE: This study evaluated the effects of different auxiliary preparation features on the resistance form of crowns with reduced axial wall and total occlusal convergence. MATERIAL AND METHODS: An Ivorine tooth was prepared on a milling machine with 20-degree total occlusal convergence (TOC), 2.5 mm of occlusocervical dimension, and a shoulder finish line. This design lacked geometric resistance form. The crown preparation was subsequently modified to include mesiodistal grooves, mesiodistal boxes, buccolingual grooves, occlusal inclined planes, an occlusal isthmus, and reduced TOC in the axial wall from 20 to 8 degrees TOC in the cervical 1.5 mm of the axial wall. The grooves and boxes were placed into the tooth with the same 20-degree TOC as the initial axial walls. Ten standardized metal dies were used for each preparation design. Standardized complete metal crowns were fabricated for all specimens. The metal crowns were cemented on metal dies with resin-modified glass ionomer cement. A strain gauge was placed at the mid-lingual cervical area of each crown preparation margin. The resistance of each specimen was evaluated when force was applied at a 45-degree angulation to the long axis of the die in a lingual to buccal direction. The peak loads during crown dislodgment, as well as the tensile stress at the mid-lingual cervical area, were measured using a universal testing machine (Kgs) for each specimen. The control group consisted of 10 dies, with the original crown preparation having no geometric resistance form and no auxiliary preparation features. Strain gauges provided the force (Kgs) that resulted in electric currency disrupt at the crown/die interface, thus providing data regarding the force required for slight crown micromovement (2 microm). Data between control and experimental groups were compared using the Mann-Whitney U test (alpha=.05). RESULTS: Proximal grooves, proximal boxes, buccolingual grooves, occlusal inclined planes, and occlusal isthmuses were not effective at increasing a crown's resistance to dislodgement when the tooth preparation lacked resistance. The only crown modification that offered enhanced resistance form when compared with the control group was the reduced TOC in the cervical half of the axial wall. CONCLUSION: Within the limitations of this in vitro study the crown preparation modification that significantly enhanced the resistance form of a compromised tooth preparation was reducing the TOC at the cervical aspect of the axial wall. Placing auxiliary retentive features such as grooves and boxes into a compromised tooth preparation (2.5 mm occlusocervical dimension and 20 degree TOC) was not effective when these retentive features possessed the same 20 degree TOC as the prepared axial walls. PMID- 14739892 TI - Microleakage of Class V resin composite restorations placed with self-etching adhesives. AB - STATEMENT OF PROBLEM: Although they are widely available, there is insufficient information about the capability of self-etching adhesives in sealing the margins of resin composite restorations. PURPOSE: The purpose of this study was to compare the in vitro microleakage of Class V resin composite restorations placed using a strong pH self-etching adhesive, an intermediate pH self-etching adhesive, and an adhesive with a separate etchant and primer. MATERIAL AND METHODS: Class V resin composite restorations (n = 8) were placed in prepared cavities in extracted human third molars using 2 self-etching dentin adhesives, Prompt L-Pop (Strong pH) or One-Up Bond F (Intermediate pH), and an adhesive with a separate etchant and primer, ScotchBond Multi-Purpose. The restored teeth received 1000 thermal cycles between 5 degrees C and 55 degrees C water baths with a 1-minute dwell time and were subsequently subjected to a methylene blue dye challenge and sectioned. The sectioned specimens were scored as demonstrating none, slight, or severe leakage (n=16). Ranked data were analyzed using a 1-way analysis of variance at a 5% confidence level. RESULTS: The only leakage observed was along gingival margins, with an incidence of 31% for Prompt L-Pop and One-Up Bond F, and 50% for ScotchBond Multi-Purpose. CONCLUSION: No significant differences in marginal leakage were found among the adhesives tested. PMID- 14739893 TI - Effects of surface treatment and joint shape on microtensile bond strength of reattached root dentin segments. AB - STATEMENT OF PROBLEM: An effective bonding technique for reattaching vertical root fracture segments has not yet been established. PURPOSE: The purpose of this study was to investigate the effects of surface treatment and joint shape on bond strength of reattached root dentin segments. MATERIAL AND METHODS: Microtensile dentin specimens (n=120, cross-sectional area=1.5 mm2) were obtained from 12 bovine teeth roots, and fractured under a tensile force. The segments of each group (n=6) were reattached by a 4-META/MMA-TBB adhesive resin after different treatments (etching or polishing, a 50-microm or 500-microm gap space, butt or wedge joint). The group with a 50-microm gap space and butt joint, which was not etched or polished, served as the control. After 24 hours of storage in 0.9% normal saline solution or thermal cycling (3000 cycles, 4 degrees C and 60 degrees C), the specimens were fractured in a universal testing machine at a crosshead speed of 1.0 mm/min. The microtensile bond strengths (MPa) were recorded. The bonded interface and fractured surface were observed by SEM (original magnification x 1000). The results were analyzed by a 2-way analysis of variance and the Fisher protected least significant difference post-hoc test (alpha=.05). RESULTS: Significantly higher bond strengths (P=.0001) were shown for the nonpolished group (9.78 +/- 4.44 MPa) and the etched group (9.31 +/- 1.23 MPa) than for the polished group (fractured before test). The wedge-shaped joint exhibited the highest bond strength (18.57 +/- 3.40 MPa, P=.0017). SEM observations showed that the penetration of resin tags along the oblique dentinal tubules was within the wedge joint. CONCLUSION: Etching and oblique tubule orientation contributed to significantly high bond strength of reattached segments to dentin after vertical root fracture. PMID- 14739894 TI - Pre-extraction records for complete denture fabrication: a literature review. AB - Dentists use arbitrary methods in determining the vertical dimension of occlusion, and arranging the maxillary anterior teeth. Some dentists have difficulty in making maxillomandibular records for completely edentulous patients. Therefore the use of pre-extraction records has been recommended. This article reviews the pre-extraction records proposed for determining the vertical dimension of occlusion, recording centric relation, and arranging the maxillary anterior teeth. A MEDLINE search, from 1966-2002, for the key words "pre extraction records" was supplemented with a hand search to identify relevant peer reviewed English articles published in dental journals. On the basis of the review, it appeared that pre-extraction records proposed for determining the original vertical dimension of occlusion and arranging the maxillary anterior teeth for a completely edentulous patient were useful and preferred to arbitrary methods, whereas pre-extraction records proposed for recording centric relation may not be necessary. PMID- 14739895 TI - A comparison of the accuracy of two methods used by pre-doctoral students to measure vertical dimension. AB - STATEMENT OF PROBLEM: Measuring vertical dimension is a soft-tissue measurement. Therefore, inaccuracy may occur. PURPOSE: The purpose of this study is to compare the accuracy of the Willis gauge method with the caliper method. MATERIALS AND METHODS: The Willis gauge measures the distance between the septum of the nose and the chin. The caliper method measures the distance between reference points on the tip of the nose and the chin. Twenty predoctoral students applied both methods 10 times in measuring the rest vertical dimension (RVD) and the occlusal vertical dimension (OVD) of a single edentulous patient. The measurements obtained from one experienced clinician were selected as controls for the interocclusal distances (IOD) for the Willis and the caliper methods, respectively. One-sided t tests and a 1-sided nonparametric test were used to determine significant differences between the 2 methods (alpha=.05). RESULTS: The variances in the RVD values for the Willis gauge method were higher than for the caliper method for most students. A Wilcoxon signed rank test showed that the accuracy of the OVD measurements for the caliper method was significantly better than for the Willis gauge method (P=.001). This was not the case for the RVD measurements (P=.073). The average IOD for the Willis method was significantly higher than the control IOD (P=.026). The average IOD for the caliper method was not significantly larger than the control (P=.1303). CONCLUSION: This study showed that the use of the caliper method by predoctoral students was a significantly more reliable method of measuring the OVD for the patient evaluated. PMID- 14739896 TI - The efficacy of palatal augmentation prostheses for speech and swallowing in patients undergoing glossectomy: a review of the literature. AB - This article provides a review of the literature to assist the clinician in determining the efficacy of palatal augmentation prosthesis regarding speech and swallowing for the patient undergoing glossectomy. A MEDLINE search was conducted. Peer-reviewed articles published from 1966 to July 2002 that addressed the question of the efficacy of this prosthesis to improve speech and swallowing after partial or total glossectomy were included. Studies reviewed had to incorporate an objective evaluation of one or both functions. Nine of 130 studies met the selection criteria (4 retrospective, 1 case control, and 4 case reports). A total of 50 subjects were studied, 42 for swallowing and 37 for speech. In 36/42 subjects, treatment was advantageous for swallowing and in 32/37 subjects, it was advantageous for speech. On the basis of the limited evidence available, the functional efficacy of the palatal augmentation prosthesis is supported. PMID- 14739897 TI - Obturation of the partial soft palate defect. AB - A partially resected soft palate represents a more significant challenge for effective prosthodontic obturation than for complete soft palate resections. In fact, for some patients it may not be possible to prosthetically rehabilitate with partial soft palate resections, resulting in velopharyngeal dysfunction, which could include hypernasal voice quality and nasal regurgitation of food and liquids. This article presents a technique for recording the contours of a partial soft-palate defect for prosthetic obturation. PMID- 14739898 TI - Optical modeling of extraoral defects. AB - In order to reduce the stress caused to patients by conventional methods of modeling using computed tomography (CT) or magnetic resonance imaging (MRI), an optical modeling process has been developed for extraoral defects and body areas. The selected body part is digitized using optical 3-coordinate measuring technology, providing an extensive data record. This is adapted for further use by equalizing the point clouds to obtain a Computer Aided Design (CAD) model, which is converted to a physical model by means of a stereolithographic process. With this technology, the patient's physical and psychological stress may be reduced. This article describes a technique for optical modeling of an ocular prosthesis. PMID- 14739899 TI - A technique for fabricating patterns for removable partial denture frameworks using digitized casts and electronic surveying. AB - Although computer-aided design and manufacture techniques have shown some promising applications in the fabrication of crowns, inlays, and maxillofacial and oral surgery, the field of removable prosthodontics has not embraced these technologies so far. This article describes the development and investigation of computer-aided techniques that may eventually enable prosthodontic procedures such as surveying and the production of sacrificial patterns to be performed digitally. A 3-dimensional computer model of a conventional cast from a patient was obtained using an optical surface capture device (a scanner). The shape of a number of components of a removable partial denture framework was modeled on the 3-dimensional scan electronically, using computer-aided design software. A physical plastic shape of the components was produced using a Rapid Prototyping machine and used as a sacrificial pattern. Techniques to allow digital cast surveying before the production of sacrificial patterns were also developed. The results show that digital dental surveying and machine-produced sacrificial patterns can be accomplished. This article forms a basis for further developments leading to a fully integrated approach to the computer-aided design and fabrication of removable partial denture frameworks. PMID- 14739900 TI - Use of carbon-epoxy frameworks for reinforcing provisional fixed partial dentures. AB - This article describes a technique for reinforcing provisional fixed partial dentures. Solid carbon-epoxy rods were shaped and coated with opaque resin to strengthen them for PMMA restorations of 3 or more units with at least 1 pontic. Because of the rigid nature of the prepolymerized carbon-epoxy material, only straight fixed partial dentures can be fabricated with this technique that allows for simplified fabrication of reinforced metal-free and cost-effective restorations. PMID- 14739901 TI - A double casting technique to minimize distortion when constructing fixed partial dentures on implants. AB - The casting of extensive implant superstructures that have a good passive fit may be technically demanding. A simple, 2-stage casting technique is presented that avoids common problems associated with the casting of large superstructures. The fabrication of a 7-unit metal-ceramic screw-retained fixed partial denture supported by 5 implants is used to illustrate this technique. PMID- 14739902 TI - A locating splint for placing implant abutments. PMID- 14739904 TI - Altered E-cadherin and epidermal growth factor receptor expressions are associated with patient survival in lung cancer: a study utilizing high-density tissue microarray and immunohistochemistry. AB - E-cadherin (E-cad) and epidermal growth factor receptor (EGFR) are important cell adhesion and signaling pathway mediators. This study aimed to assess their expression in lung adenocarcinoma (AdC) and squamous cell carcinoma (SCC) and their association with clinicopathologic variables. In all, 130 resectable lung cancers (stages I-IIIA) were studied using a high-density tissue microarray. Two to three cores from each case were arrayed into three blocks using a Beecher system. Immunohistochemistry was performed using an avidin-biotin complex method and monoclonal antibodies against E-cad and EGFR. Unequivocal membrane staining in >10% of tumor cells was considered as a positive expression of E-cad and EGFR. Markers expression and coexpression were analyzed against clinicopathologic variables (age, gender, smoking status, performance status, weight loss, histology, grade, stage, and lymph node involvement) and patient survival. There were 118, 126, and 115 cases that were fully assessable for E-cad, EGFR, and both markers, respectively. For E-cad, 65 cases (55%) were positive (+), 53 (45%) were negative (-); 23 cases of the negative group had only cytoplasmic staining. For EGRF, 43 cases (34%) were (+), and 83 (66%) were (-). There was no significant association between E-cad or EGFR, and any of the clinicopathologic variables except for an association between EGFR(+) and SCC histologic type. Both negative and cytoplasmic staining of E-cad correlated with shorter patient survival with P=0.008 and 0.002, respectively. EGFR expression did not correlate with patient survival; however, patients with E-cad(-)/EGFR(+) phenotype had poorer survival than those with E-cad(+)/EGFR(-) (P=0.026). Our study suggests that lung AdC and SCC may be stratified based on expression of E-cad and EGFR with the E-cad( )/EGFR(+) expression having a worse disease outcome. Moreover, the cytoplasmic expression of E-cad may represent an altered localization of this protein in association with tumorigenicity. PMID- 14739905 TI - Diagnosis and reporting of limited adenocarcinoma of the prostate on needle biopsy. AB - The diagnosis of limited adenocarcinoma of the prostate is one of the more difficult challenges in surgical pathology. This paper highlights the methodological approach to diagnosing limited cancer, based on a constellation of features more commonly present in adenocarcinoma than benign glands. In assessing small foci of atypical glands on needle biopsy, one looks for differences between the benign glands and the atypical glands in terms of nuclear features, cytoplasmic features, and intraluminal contents. Only a few features, such as glomerulations, mucinous fibroplasia (collagenous micronodules), and perineural invasion are diagnostic in and of themselves for prostate cancer. Immunohistochemistry may be a useful adjunct in the diagnosis of limited adenocarcinoma of the prostate, although as with any immunohistochemical studies, there are problems with both sensitivity and specificity. Basal cell markers, such as high molecular weight cytokeratin and more recently, p63, highlight basal cells found in benign glands, yet are absent in adenocarcinoma of the prostate. However, not all benign glands label uniformly with basal cell markers. Certain mimickers of adenocarcinoma of the prostate are even less frequently labeled uniformly with these stains. Consequently, negative staining in a small focus of atypical glands for basal cell markers is not diagnostic of adenocarcinoma of the prostate. More recently, a marker has been identified that relatively selectively labels adenocarcinoma of the prostate. AMACR will label the cytoplasm of approximately 80% of limited adenocarcinoma of the prostate cases on needle biopsy. In positive cases, not all of the glands will be positive and those that are positive are often not intensely positive. Certain variants of adenocarcinoma of the prostate that are a little more difficult to recognize, such as foamy glands adenocarcinoma, pseudohyperplastic adenocarcinoma, and atrophic adenocarcinoma, are labeled with AMACR in only approximately 60-70% of cases. In addition to problems with sensitivity, AMACR is not entirely specific for adenocarcinoma, and will label almost all cases of high-grade prostatic intraepithelial neoplasia, some foci of adenosis, and even some entirely benign glands. Finally, this paper will briefly cover the significance of atypical or suspicious prostate needle biopsies, and how to report the key diagnostic and prognostic information on needle biopsy. PMID- 14739906 TI - High-grade prostatic intraepithelial neoplasia. AB - High-grade prostatic intraepithelial neoplasia (PIN) is now accepted as the most likely preinvasive stage of adenocarcinoma, almost two decades after its first formal description. PIN has a high predictive value as a marker for adenocarcinoma, and its identification warrants repeat biopsy for concurrent or subsequent invasive carcinoma. The only method of detection is biopsy; PIN does not significantly elevate serum prostate-specific antigen (PSA) concentration or its derivatives and cannot be detected by current imaging techniques, including ultrasound. Most patients with PIN will develop carcinoma within 10 years. PIN is associated with progressive abnormalities of phenotype and genotype, which are similar to cancer rather than normal prostatic epithelium, indicating impairment of cell differentiation with advancing stages of prostatic carcinogenesis. Androgen deprivation therapy decreases the prevalence and extent of PIN, suggesting that this form of treatment may play a role in chemoprevention. PMID- 14739907 TI - LIFE and SCOPE: what guidance do they offer alongside ALLHAT and ANBP2? PMID- 14739908 TI - Blood pressure and pulse responses to three stressors: associations with sociodemographic characteristics and cardiovascular risk factors. AB - Cardiovascular reactivity is hypothesized to increase the risk of hypertension and other CVD-related conditions. However, studies to date are inconclusive. We compared the association of blood pressure and pulse responses to three stressors (postural challenge, handgrip test, mental arithmetic) with sociodemographic characteristics and CVD risk factors. We included 782 participants from the Hypertension Genetic Epidemiology Study. Blood pressure and pulse responses to stressors were defined as the difference between post- and pre-stress measurements. Stepwise regression analyses examined change in SBP and pulse in response to stressors as a function of sociodemographic and CVD risk factors. Age, race, and gender were forced into models and other variables (education, BMI, waist circumference, resting SBP and DBP, cigarette smoking, LDL and HDL cholesterol, glucose, and antihypertensive medications (beta-blockers, calcium channel blockers, diuretics, ace inhibitors)) were retained if P<0.10. Age was a significant predictor of SBP response to all stressors. The SBP response to a change in posture was not related to other variables. The SBP response to mental arithmetic was significantly higher among men, those with larger waists, higher SBP, beta-blocker users, and lower among smokers. SBP response to the handgrip was significantly higher among those with higher SBP and beta-blocker users. Similarly, the association of the pulse response to the risk factors varied considerably across the stressors. Overall, the socio-demographic and CVD risk factors accounted for between 9 and 14% of the variance in the SBP response to the stressors and from between 4 and 12% of the variance in the pulse response to the three stressors. The associations between sociodemographic and CVD risk factors and the SBP and pulse response to stress were modest and inconsistent across stressors. The findings suggest that cardiovascular reactivity is a concept that needs to be defined in reference to specific stressors so that mechanisms leading to responses can be better understood. PMID- 14739909 TI - Alcohol consumption, homeostasis model assessment indices and blood pressure in middle-aged healthy men. AB - A total of 1595 middle-aged healthy men consuming alcohol up to 120 ml per day and 538 without alcohol consumption were recruited from an occupational population, and their insulin resistance (IR) and beta-cell function (BC) were measured using the homeostasis model assessment (HOMA-IR and HOMA-BC), and the associations with alcohol consumption, blood pressure (BP), and serum gamma glutamyltransferase (GGT) levels were analysed cross-sectionally. Both HOMA-IR and HOMA-BC were decreased with increasing alcohol consumption, but HOMA-BC corresponding to a level of HOMA-IR was 4-10 and 8-20% lower in drinkers consuming less than 60 ml of alcohol per day and those consuming more, respectively, than in nondrinkers, suggesting an altered fasting serum insulin glucose relationship in alcohol consumers. Although BP was higher and HOMA-IR was lower in alcohol consumers than in nonconsumers, BP was higher at higher HOMA-IR irrespective of alcohol consumption. Elevations of serum GGT were positively associated with BP and HOMA-IR in both alcohol consumers and nonconsumers. Multiple regression analyses in the subjects showed that elevated serum GGT was an independent contributor to HOMA-IR elevations, and both serum GGT and HOMA-IR were significantly related to BP elevations after adjusting for alcohol consumption, age, body mass index, cigarette consumption, and physical activity at leisure. Although cross-sectional observations do not provide evidence of causal association, the results suggest that elevated serum GGT in alcohol consumers relates to elevations of IR and that the elevated insulin resistance relates, at least partly, to BP elevations in alcohol consumers. PMID- 14739911 TI - Traumatic superior orbital fissure syndrome. PMID- 14739912 TI - Cataract surgery in latex allergy patients. PMID- 14739913 TI - Angiographic pattern of recurrent choroidal neovascularization in age-related macular degeneration. AB - PURPOSE: To evaluate the angiographic characteristics of recurrent choroidal neovascularization (R-CNV) in age-related macular degeneration (AMD). METHODS: A prospective investigation on 107 consecutive patients with exudative AMD and CNV not involving the fovea was conducted. Fluorescein angiography (FA) and indocyanine green angiography (ICGA) were planned before krypton laser treatment, and after 3 weeks, 2, 3, 4, 6, 9, 12, 18, and 24 months from photocoagulation. Laser treatment was FA-guided in eyes with classic CNV, and ICGA-guided in eyes with occult CNV on FA. RESULTS: At baseline on FA, 23.3% had classic CNV, whereas, 76.6% showed occult CNV. On ICGA, CNV assumed a focal and a plaque pattern in 81.3 and 18.6% of cases, respectively. Overall, post-laser CNVs occurred in 56 eyes. FA identified well-defined and ill-defined R-CNV in 25 and 75% of cases, respectively. ICGA identified three different R-CNV patterns: focal, annular, and plaque. Focal R-CNV was defined as a single dot-like hyperfluorescence, which was detected in 69.6% of cases, with subfoveal location in half of them. Annular R-CNV was identified by a hyperfluorescent lesion, partially or completely encircling treated area, which was visible in 19.6% of cases, all with subfoveal involvement. Plaque R-CNV was defined as a hyperfluorescent lesion larger than 1 disc diameter in size, and was seen in 10.7% of cases, all with subfoveal location. CONCLUSIONS: ICGA is able to improve R-CNV visualization identifying three different R-CNV patterns. Focal R-CNV is the most frequent pattern and can be re-treated in half of the cases. PMID- 14739914 TI - Periorbital oedema and epiphora as ocular side effects of imatinib mesylate (Gleevec). PMID- 14739915 TI - Detection of oestrogen receptors (ER) alpha and beta in conjunctiva, lacrimal gland, and tarsal plates. AB - PURPOSE: Lack of Oestrogen and androgen may be of importance in the pathogenesis of keratoconjunctivitis sicca (KCS). The signal of Oestrogens is transmitted via specific Oestrogen receptors (ER). It was the purpose of this study to evaluate the expression of ER alpha and ER beta in tear-producing tissues. METHODS: Reverse transcriptase-polymerase chain reaction (RT-PCR, ER alpha + beta) and immunohistochemical evaluation (ER alpha only) were performed for ER detection and localization in tissue samples of bulbar conjunctiva (20 samples of 20 patients undergoing cataract surgery), tarsal plates (14 samples of 12 patients undergoing eye lid surgery), and lacrimal glands (20 samples of 13 cornea donors). RESULTS: Messenger RNA ER alpha was identified via RT-PCR in all tissue samples with variable expression, ER beta predominantly in lacrimal gland tissue. Immunohistochemical staining for ER alpha was negative in most cases, probably due to the thermolability of ERs and very small sample sizes. CONCLUSIONS: The detection of ER alpha and ER beta mRNA expression supports the concept of a receptor-based effect of Oestrogen in these tissues contributing to KCS. This may encourage therapeutical efforts including topical Oestrogen administration. PMID- 14739916 TI - Iatrogenic macula hole and consequent macular detachment caused by intravitreal trypan blue injection. PMID- 14739917 TI - Subluxation of intraocular lens within the capsular bag following a contusion injury. PMID- 14739918 TI - Bilateral scotomas associated with retinal hemorrhages following endoscopic spinal surgery. PMID- 14739919 TI - Penetrating keratoplasty in adults with congenital glaucomas. AB - PURPOSE: To report the problems and outcomes of penetrating keratoplasty (PKP) in patients with corneal oedema due to congenital glaucoma. METHOD: Case note review of nine such consecutive patients treated with PKP. A Kaplan - Meir survival curve was plotted for the series. RESULTS: Nine patients, ranging in age from 27 to 71 years at the time of their surgery were followed up for a mean of 28 months. All had undergone previous surgery for glaucoma. Preoperatively all patients had controlled intraocular pressures, and four were on ocular hypotensives. Two patients developed corneal graft failure at 15 and 41 months postoperative. Raised pressure was the cause in both. Final visual acuity improved in five patients, was the same in three, and deteriorated in only one. Complications included raised intraocular pressure, cataract, retinal detachment, and endophthalmitis secondary to an infected Molteno implant. CONCLUSION: Despite multiple impediments to good postoperative vision including previous surgery and pre-existing glaucomatous damage, a successful outcome was achieved by careful patient selection, and an awareness of, and meticulous attention to postoperative complications. PMID- 14739920 TI - Delayed recurrence of ciliary body melanoma following local resection. PMID- 14739921 TI - Intramuscular lipoma of the eyelid: a case report. PMID- 14739922 TI - Keratocyte cytotoxicity of riboflavin/UVA-treatment in vitro. AB - PURPOSE: Collagen crosslinking using ultraviolet- A (UVA) -irradiation combined with the photosensitizer riboflavin is a new technique for treating progressive keratoconus. It has been shown to increase effectively the biomechanical strength of the cornea and to stop or even reverse the progression of keratoconus. As part of a safety evaluation, the present study was undertaken to investigate in vitro the possible cytotoxic effect of combined riboflavin/UVA-treatment on corneal keratocytes and to compare it to UVA-irradiation alone. METHODS: Cell cultures established from porcine keratocytes were treated with 0.025% riboflavin solution and various UVA (370 nm)-irradiances ranging from 0.4 to 1.0 mW/cm2 and with UVA alone between 2 and 9 mW/cm2 for 30 min. The cell cultures were evaluated for cell death 24 h after irradiation using trypan-blue and Yopro-fluorescence staining. RESULTS: An abrupt cytotoxic irradiance level was found at 0.5 mW/cm2 for keratocytes after UVA-irradiation combined with the photosensitizer riboflavin, which is 10-fold lower than the cytotoxic irradiance of 5 mW/cm2 after UVA-irradiation alone. CONCLUSIONS: A cytotoxic effect of combined riboflavin/UVA-treatment on keratocytes is to be expected at 0.5 mW/cm2, which is reached in the clinical setting in human corneas down to a depth of 300 microm using the standard surface UVA-irradiance of 3 mW/cm2. PMID- 14739923 TI - Diurnal variation in microcirculation of ocular fundus and visual field change in normal-tension glaucoma. AB - PURPOSE: Diurnal variations in microcirculation of the ocular fundus in normal tension glaucoma (NTG) were examined to compare with the normal control eyes. The correlation between progression of visual field impairment and diurnal variations in ocular circulation was also studied. METHODS: The subjects were 12 patients with NTG and 12 normal controls. Blood pressure (BP), intraocular pressure (IOP), ocular perfusion pressure (OPP), and square blur rate (SBR), an index of microcirculation acquired by the laser speckle method, were measured at 9 a.m. (morning), 3 p.m. (afternoon), and 9 p.m. (night). Diurnal variations in SBR were tested using the Friedman test and Wilcoxon signed ranks test. On the other hand, diurnal variations in BP, IOP, and OPP were tested by the analysis of variance. The visual field was evaluated at the same time as determining diurnal variation and again about 9 months later to calculate the change. The correlation between variation ratio in SBR and the change in visual field was examined by simple regression. RESULTS: A significant decrease (P=0.04) was found in SBR at night, as compared with the morning value, in the optic nerve head (ONH) of NTG, although no significant diurnal variations were found in SBR either in the choroid-retina or in normal control eyes. Other parameters showed no significant diurnal variations. The larger diurnal variation was in SBR of the ONH, and the more exacerbated visual field impairment was (r=0.59, P=0.04). CONCLUSION: These findings suggest that diurnal variations in the microcirculation of the ONH may play a role in the progression of NTG. PMID- 14739924 TI - Subnormal visual perception in school-aged ex-preterm patients in a paediatric eye clinic. AB - PURPOSE: The aim of this study was to assess visual perception at school age of children born preterm with known lesions to the posterior visual pathways or with ophthalmologic signs that might indicate such lesions. METHODS: The study group consisted of 91 patients born before the 37th gestational week. Visual perception was assessed using the TVPS-R (Test of Visual Perceptual Skills - Revised) and a structured interview. In addition, ophthalmologic and orthoptic examinations were performed. RESULTS: On the test of visual perception, 67% of the patients had results below the third percentile of the American reference group. This is to be compared with 10% of Swedish full-term controls. Scores below the third percentile were observed in 87% of the patients with known brain lesions, 48% of those with strabismus without known brain lesion, and 86% of those with reduced visual acuity in the absence of strabismus and known brain lesion. CONCLUSIONS: Reduced visual perception is common among children born preterm who have strabismus and/or reduced visual acuity, as well as in those with known brain lesions. This study emphasises the need to find tools to identify and assess those patients who have visual perceptual problems that may restrict their ability to meet the demands of daily life. PMID- 14739925 TI - Extrafoveal seafan neovascularization associated with unilateral recurrent retinal capillary haemangiomas: a known association or a mere coincidence. PMID- 14739926 TI - Transpupillary thermotherapy for treatment of exudative age-related macular degeneration in Japanese patients. AB - PURPOSE: To report the therapeutic outcome of transpupillary thermotherapy (TTT) for subfoveal choroidal neovascularizarion (CNV) in brown retina using a diode laser with the setting of lower energy level compared to the previous studies on light-pigmented Caucasian patients. METHODS: A total of 19 subfoveal CNVs in 18 patients were treated with TTT. The power of diode-laser was set 160 mW for 1.2 mm beam, 270 mW for 2.0 mm beam, and 400 mW for 3.0 mm beam, and the laser was delivered for 1 min through a slit-lamp mounted-delivery system. Patients were followed up for a mean of 8.8 months (4-12 months). Visual acuity and the fundus change as judged by funduscopic examination and simultaneous fluorescein and indocyanine green angiography were evaluated. Visual acuity was measured by a Japanese standard Landolt visual acuity chart and converted to logarithm of the minimal angle resolution (log MAR) visual acuity for statistical analysis. Improvement or decline in vision was defined as change of more than 0.2 in log MAR visual acuity. RESULTS: In eyes with minimally classic or occult only CNV, visual acuity improved in two eyes (18%) stabilized in seven eyes (64%) and worsened in two eyes (18%). In eyes with predominantly classic CNV, visual acuity improved in two eyes (25%), stabilized in four eyes (50%) and worsened in two eyes (25%). In all, 15 (84%) eyes of all studied subjects had improvement in exudation. Two (11%) and one (5%) eye(s) were noted to have a significant post treatment haemorrhage and retinal pigment epithelial tear, respectively. CONCLUSION: In patients with brown retinal colour, the treatment outcome of TTT was comparable to that of light-pigmented Caucasian patients with approximately half the laser power energy. Further randomized control studies are warranted. PMID- 14739927 TI - Regulation of checkpoint kinases through dynamic interaction with Crb2. AB - ATR/Rad3-like kinases promote the DNA damage checkpoint through regulating Chk1 that restrains the activation of cyclin-dependent kinases. In fission yeast, Crb2, a BRCT-domain protein that is similar to vertebrate 53BP1, plays a crucial role in establishing this checkpoint. We report here that Crb2 regulates DNA damage checkpoint through temporal and dynamic interactions with Rad3, Chk1 and replication factor Cut5. The active complex formation between Chk1 and Crb2 is regulated by Rad3 and became maximal during the checkpoint arrest. Chk1 activation seems to need two steps of interaction changes: the loss of Rad3-Chk1 and Rad3-Crb2 interactions, and the association between hyperphosphorylated forms of Chk1 and Crb2. Chk1 is the major checkpoint kinase for the arrest of DNA polymerase mutants. The in vitro assay of Chk1 showed that its activation requires the presence of Crb2 BRCT. Hyperphosphorylation of Crb2 is also dependent on its intact BRCT. Finally, we show direct interaction between Rad3 and Crb2, which is inhibitory to Rad3 activity. Hence, Crb2 is the first to interact with both Rad3 and Chk1 kinases. PMID- 14739928 TI - Nhp6 facilitates Aft1 binding and Ssn6 recruitment, both essential for FRE2 transcriptional activation. AB - We found Nhp6a/b yeast HMG-box chromatin-associated architectural factors and Ssn6 (Cyc8) corepressor to be crucial transcriptional coactivators of FRE2 gene. FRE2 encoding a plasma membrane ferric reductase is induced by the iron responsive, DNA-binding, transcriptional activator Aft1. We have shown that Nhp6 interacts directly with the Aft1 N-half, including the DNA-binding region, to facilitate Aft1 binding at FRE2 UAS. Ssn6 also interacts directly with the Aft1 N half and is recruited on FRE2 promoter only in the presence of both Aft1 and Nhp6. This Nhp6/Ssn6 role in Aft1-mediated transcription is FRE2 promoter context specific, and both regulators are required for activation-dependent chromatin remodeling. Our results provide the first in vivo biochemical evidence for nonsequence-specific HMG-box protein-facilitated recruitment of a yeast gene specific transactivator to its DNA target site and for Nhp6-mediated Ssn6 promoter recruitment. Ssn6 has an explicitly coactivating role on FRE2 promoter only upon induction. Therefore, transcriptional activation in response to iron availability involves multiple protein interactions between the Aft1 iron responsive DNA-binding factor and global regulators such as Nhp6 and Ssn6. PMID- 14739929 TI - Crystal structures of histone Sin mutant nucleosomes reveal altered protein-DNA interactions. AB - Here we describe 11 crystal structures of nucleosome core particles containing individual point mutations in the structured regions of histones H3 and H4. The mutated residues are located at the two protein-DNA interfaces flanking the nucleosomal dyad. Five of the mutations partially restore the in vivo effects of SWI/SNF inactivation in yeast. We find that even nonconservative mutations of these residues (which exhibit a distinct phenotype in vivo) have only moderate effects on global nucleosome structure. Rather, local protein-DNA interactions are disrupted and weakened in a subtle and complex manner. The number of lost protein-DNA interactions correlates directly with an increased propensity of the histone octamer to reposition with respect to the DNA, and with an overall destabilization of the nucleosome. Thus, the disruption of only two to six of the approximately 120 direct histone-DNA interactions within the nucleosome has a pronounced effect on nucleosome mobility and stability. This has implications for our understanding of how these structures are made accessible to the transcription and replication machinery in vivo. PMID- 14739930 TI - Transitions in RNA polymerase II elongation complexes at the 3' ends of genes. AB - To understand the factor interactions of transcribing RNA polymerase II (RNApII) in vivo, chromatin immunoprecipitations were used to map the crosslinking patterns of multiple elongation and polyadenylation factors across transcribed genes. Transcription through the polyadenylation site leads to a reduction in the levels of the Ctk1 kinase and its associated phosphorylation of the RNApII C terminal domain. One group of elongation factors (Spt4/5, Spt6/Iws1, and Spt16/Pob3), thought to mediate transcription through chromatin, shows patterns matching that of RNApII. In contrast, the Paf and TREX/THO complexes partially overlap RNApII, but do not crosslink to transcribed regions downstream of polyadenylation sites. In a complementary pattern, polyadenylation factors crosslink strongly at the 3' ends of genes. Mutation of the 3' polyadenylation sequences or the Rna14 protein causes loss of polyadenylation factor crosslinking and read-through of termination sequences. Therefore, transcription termination and polyadenylation involve transitions at the 3' end of genes that may include an exchange of elongation and polyadenylation/termination factors. PMID- 14739931 TI - Phosphorylation of MRF4 transactivation domain by p38 mediates repression of specific myogenic genes. AB - Skeletal myogenesis is associated with the activation of four muscle regulatory factors (MRFs): Myf5, MyoD, Myogenin and MRF4. Here we report that p38 mitogen activated protein kinase represses the transcriptional activity of MRF4 (involved in late stages of myogenesis), resulting in downregulation of specific muscle genes. MRF4 is phosphorylated in vitro and in vivo by p38 on two serines (Ser31 and Ser42) located in the N-terminal transactivation domain, resulting in reduced MRF4-mediated transcriptional activity. In contrast, nonphosphorylatable MRF4 mutants display increased transcriptional activity and are able to advance both myoblast fusion and differentiation. We also show that expression of desmin and alpha-actin, but not muscle creatin kinase, decreased at late stages of muscle differentiation, correlating with the induction of MRF4 and p38 activation. Accordingly, inhibition of p38 during late myogenesis results in the upregulation of both desmin and alpha-actin. We propose that repression of MRF4 activity by p38 phosphorylation may represent a new mechanism for the silencing of specific muscle genes at the terminal stages of muscle differentiation. PMID- 14739932 TI - Dopamine modulates the plasticity of mechanosensory responses in Caenorhabditis elegans. AB - Dopamine-modulated behaviors, including information processing and reward, are subject to behavioral plasticity. Disruption of these behaviors is thought to support drug addictions and psychoses. The plasticity of dopamine-mediated behaviors, for example, habituation and sensitization, are not well understood at the molecular level. We show that in the nematode Caenorhabditis elegans, a D1 like dopamine receptor gene (dop-1) modulates the plasticity of mechanosensory behaviors in which dopamine had not been implicated previously. A mutant of dop-1 displayed faster habituation to nonlocalized mechanical stimulation. This phenotype was rescued by the introduction of a wild-type copy of the gene. The dop-1 gene is expressed in mechanosensory neurons, particularly the ALM and PLM neurons. Selective expression of the dop-1 gene in mechanosensory neurons using the mec-7 promoter rescues the mechanosensory deficit in dop-1 mutant animals. The tyrosine hydroxylase-deficient C. elegans mutant (cat-2) also displays these specific behavioral deficits. These observations provide genetic evidence that dopamine signaling modulates behavioral plasticity in C. elegans. PMID- 14739933 TI - Hfq, a new chaperoning role: binding to messenger RNA determines access for small RNA regulator. AB - The Sm-like protein Hfq is involved in post-transcriptional regulation by small, noncoding RNAs in Escherichia coli that act by base pairing. Hfq stabilises the small RNAs and mediates their interaction with the target mRNA by an as yet unknown mechanism. We show here a novel chaperoning use of Hfq in the regulation by small RNAs. We analysed in vitro and in vivo the role of Hfq in the interaction between the small RNA RyhB and its sodB (iron superoxide dismutase) mRNA target. Hfq bound strongly to sodB mRNA and altered the structure of the mRNA, partially opening a loop. This gives access to a sequence complementary to RyhB and encompassing the translation initiation codon. RyhB binding blocked the translation initiation codon of sodB and triggered the degradation of both RyhB and sodB mRNA. Thus, Hfq is a critical chaperone in vivo and in vitro, changing the folding of the target mRNA to make it subject to the small RNA regulator. PMID- 14739934 TI - Plasticity in eucaryotic 20S proteasome ring assembly revealed by a subunit deletion in yeast. AB - The 20S proteasome is made up of four stacked heptameric rings, which in eucaryotes assemble from 14 different but related subunits. The rules governing subunit assembly and placement are not understood. We show that a different kind of proteasome forms in yeast when the Pre9/alpha3 subunit is deleted. Purified pre9Delta proteasomes show a two-fold enrichment for the Pre6/alpha4 subunit, consistent with the presence of an extra copy of Pre6 in each outer ring. Based on disulfide engineering and structure-guided suppressor analyses, Pre6 takes the position normally occupied by Pre9, a substitution that depends on a network of intersubunit salt bridges. When Arabidopsis PAD1/alpha4 is expressed in yeast, it complements not only pre6Delta but also pre6Delta pre9Delta mutants; therefore, the plant alpha4 subunit also can occupy multiple positions in a functional yeast proteasome. Importantly, biogenesis of proteasomes is delayed at an early stage in pre9Delta cells, suggesting an advantage for Pre9 over Pre6 incorporation at the alpha3 position that facilitates correct assembly. PMID- 14739935 TI - Structural basis for recruitment of the ATPase activator Aha1 to the Hsp90 chaperone machinery. AB - Hsp90 is a molecular chaperone essential for the activation and assembly of many key eukaryotic signalling and regulatory proteins. Hsp90 is assisted and regulated by co-chaperones that participate in an ordered series of dynamic multiprotein complexes, linked to Hsp90s conformationally coupled ATPase cycle. The co-chaperones Aha1 and Hch1 bind to Hsp90 and stimulate its ATPase activity. Biochemical analysis shows that this activity is dependent on the N-terminal domain of Aha1, which interacts with the central segment of Hsp90. The structural basis for this interaction is revealed by the crystal structure of the N-terminal domain (1-153) of Aha1 (equivalent to the whole of Hch1) in complex with the middle segment of Hsp90 (273-530). Structural analysis and mutagenesis show that binding of N-Aha1 promotes a conformational switch in the middle-segment catalytic loop (370-390) of Hsp90 that releases the catalytic Arg 380 and enables its interaction with ATP in the N-terminal nucleotide-binding domain of the chaperone. PMID- 14739936 TI - Escherichia coli YidC is a membrane insertase for Sec-independent proteins. AB - YidC is a recently discovered bacterial membrane protein that is related to the mitochondrial Oxa1p and the Alb3 protein of chloroplasts. These proteins are required in the membrane integration process of newly synthesized proteins that do not require the classical Sec machinery. Here we demonstrate that YidC is sufficient for the membrane integration of a Sec-independent protein. Microgram amounts of the purified single-spanning Pf3 coat protein were efficiently inserted into proteoliposomes containing the purified YidC. A mutant Pf3 coat protein with an extended hydrophobic region was inserted independently of YidC into the membrane both in vivo and in vitro, but its insertion was accelerated by YidC. These results show that YidC can function separately from the Sec translocase to integrate membrane proteins into the lipid bilayer. PMID- 14739938 TI - Knockdown of survivin expression by small interfering RNA reduces the clonogenic survival of human sarcoma cell lines independently of p53. AB - Survivin, a member of the inhibitors-of-apoptosis gene family, is overexpressed in many tumor types. Survivin is a prognostic marker of soft-tissue sarcomas, but the downregulation of survivin expression and the possible dependency of survivin downregulation on p53 in these tumors have not been investigated. Therefore, we applied small interfering RNA (siRNA) to knock down the expression of survivin in five human sarcoma cell lines with wild-type or mutant p53 alleles. Compared with survivin mRNA expression in the nonsense siRNA-treated sarcoma cell lines, expression after treatment with survivin-specific siRNA was reduced by 73-88%; survivin protein expression was reduced by 52-81%. This finding was coupled with a reduction in clonogenic survival ranging from 65-86%. However, less than 10% of cells treated with survivin-specific siRNA underwent apoptosis. Cell-cycle and morphologic analyses showed that after a dramatic increase in the number of treated cells in the G2/M phase, some of the cells became polyploid; this result indicates that mitosis of a substantial number of treated cells was incomplete. Our findings suggest that survivin-specific siRNA could be a selective treatment to kill sarcoma cells regardless of the presence or absence of wild-type p53 alleles. PMID- 14739937 TI - Synergy and antagonism between Notch and BMP receptor signaling pathways in endothelial cells. AB - Notch and bone morphogenetic protein signaling pathways are important for cellular differentiation, and both have been implicated in vascular development. In many cases the two pathways act similarly, but antagonistic effects have also been reported. The underlying mechanisms and whether this is caused by an interplay between Notch and BMP signaling is unknown. Here we report that expression of the Notch target gene, Herp2, is synergistically induced upon activation of Notch and BMP receptor signaling pathways in endothelial cells. The synergy is mediated via RBP-Jkappa/CBF-1 and GC-rich palindromic sites in the Herp2 promoter, as well as via interactions between the Notch intracellular domain and Smad that are stabilized by p/CAF. Activated Notch and its downstream effector Herp2 were found to inhibit endothelial cell (EC) migration. In contrast, BMP via upregulation of Id1 expression has been reported to promote EC migration. Interestingly, Herp2 was found to antagonize BMP receptor/Id1-induced migration by inhibiting Id1 expression. Our results support the notion that Herp2 functions as a critical switch downstream of Notch and BMP receptor signaling pathways in ECs. PMID- 14739939 TI - In vivo rejection of tumor cells dependent on CD8 cells that kill independently of perforin and FasL. AB - Perforin/granzyme B- and Fas/FasL-mediated killing pathways are the main effector mechanisms of CTL and NK cells in antitumor immune responses. In this study, we investigated the relative role of these two lytic mechanisms in protection of the host from tumor progression, as well as spontaneous metastasis, using the D122 Lewis lung carcinoma and its gene-modified cells. Utilizing perforin knockout mice (B6-PKO) and Fas and FasL mutant (B6-MRL and B6-Smn) mice, we found that perforin expression in the host plays a crucial function in the prevention of metastasis. However, local tumor rejection of an H-2K(b) and B7-1 transfectant, 39.5-B7 cells, was not dependent either on perforin or Fas/FasL expression in vivo. In addition, CTL lysis of 39.5-B7 cells was independent of perforin and Fas/FasL interactions in 18-hour in vitro assays. We also confirmed that CD8 T cells were responsible for rejecting 39.5-B7 local tumors, yet cytokines, TNF alpha and gammaIFN were not involved in tumor rejection in vivo. Furthermore, blocking assays using caspase inhibitors (zVAD-fmk, zLETD-fmk and zLEHD-fmk) showed that, whereas caspase activation was partially required to induce 39.5-B7 lysis mediated by the perforin-dependent pathway, 39.5-B7 lysis by CTLs through the perforin-independent mechanism required caspase activation. Thus, these results suggested that perforin, Fas/FasL, gammaIFN and TNF-alpha independent lytic mechanisms, mediated by CD8 T cells, have a crucial role in rejection of 39.5-B7 cells in vivo. Caspase activation is a pre requisite for apoptosis of targets by CTLs. PMID- 14739940 TI - Protective effects of estradiol on TRAIL-induced apoptosis in a human oligodendrocytic cell line: evidence for multiple sites of interactions. AB - Demyelinating diseases are high impact neurological disorders. Steroids are regarded as protective molecules in the susceptibility to these diseases. Here, we studied the interactions between tumour necrosis factor-related apoptosis inducing ligand (TRAIL), a potent proapoptotic molecule toxic to oligodendrocytes, and 17-beta-estradiol (E-17-beta), in human oligodendrocytic MO3.13 cells. Exposure of cells to TRAIL resulted in the upregulation of both death receptors DR4 and DR5 and apoptosis, as well as the activation of caspase-8 and -3, increased phosphorylation of Jun-N-terminal kinase and p38 kinase, and the reduction of bcl-2 and bcl-xL proteins. TRAIL-mediated MO3.13 cell apoptosis was abrogated by the dominant-negative form of the adaptor protein FADD and by caspase inhibitors. Preincubation with E-17-beta completely prevented both TRAIL induced DR4 and DR5 upregulation and apoptosis. Estrogen-induced cytoprotection was time and concentration dependent and reverted by antiestrogens. Estrogen treatment per se reduced kinase phosphorylation, and upregulated bcl-2 and bcl-xL proteins. In conclusion, our data show that the detrimental role of TRAIL on oligodendrocytes can be effectively counteracted by estrogens, thus suggesting that the underlying molecular interactions can be of potential relevance in characterizing novel targets for therapy of demyelinating disorders. PMID- 14739941 TI - CD95/phosphorylated ezrin association underlies HIV-1 GP120/IL-2-induced susceptibility to CD95(APO-1/Fas)-mediated apoptosis of human resting CD4(+)T lymphocytes. AB - CD95(APO-1/Fas)-mediated apoptosis of bystander uninfected T cells exerts a major role in the HIV-1-mediated CD4+ T-cell depletion. HIV-1 gp120 has a key role in the induction of sensitivity of human lymphocytes to CD95-mediated apoptosis through its interaction with the CD4 receptor. Recently, we have shown the importance of CD95/ezrin/actin association in CD95-mediated apoptosis. In this study, we explored the hypothesis that the gp120-mediated CD4 engagement could be involved in the induction of susceptibility of primary human T lymphocytes to CD95-mediated apoptosis through ezrin phosphorylation and ezrin-to-CD95 association. Here, we show that gp120/IL-2 combined stimuli, as well as the direct CD4 triggering, on human primary CD4(+)T lymphocytes induced an early and stable ezrin activation through phosphorylation, consistent with the induction of ezrin/CD95 association and susceptibility to CD95-mediated apoptosis. Our results provide a new mechanism through which HIV-1-gp120 may predispose resting CD4(+)T cell to bystander CD95-mediated apoptosis and support the key role of ezrin/CD95 linkage in regulating susceptibility to CD95-mediated apoptosis. PMID- 14739942 TI - Cathepsin D links TNF-induced acid sphingomyelinase to Bid-mediated caspase-9 and -3 activation. AB - Acidic noncaspase proteases-like cathepsins have been introduced as novel mediators of apoptosis. A clear role for these proteases and the acidic endolysosomal compartment in apoptotic signalling is not yet defined. To understand the role and significance of noncaspases in promoting and mediating cell death, it is important to determine whether an intersection of these proteases and the caspase pathway exists. We recently identified the endolysosomal aspartate protease cathepsin D (CTSD) as a target for the proapoptotic lipid ceramide. Here, we show that tumor necrosis factor (TNF) induced CTSD activation depends on functional acid sphingomyelinase (A-SMase) expression. Ectopic expression of CTSD in CTSD-deficient fibroblasts results in an enhanced TNF-mediated apoptotic response. Intracellular colocalization of CTSD with the proapoptotic bcl-2 protein family member Bid in HeLa cells, and the ability of CTSD to cleave directly Bid in vitro as well as the lack of Bid activation in cathepsin-deficient fibroblasts indicate that Bid represents a direct downstream target of CTSD. Costaining of CTSD and Bid with Rab5 suggests that the endosomal compartments are the common 'meeting point'. Caspase-9 and -3 activation also was in part dependent on A-SMase and CTSD expression as revealed in the respective deficiency models. Our results link as novel endosomal intermediates the A-SMase and the acid aspartate protease CTSD to the mitochondrial apoptotic TNF pathway. PMID- 14739943 TI - Tissue transglutaminase triggers oligomerization and activation of dual leucine zipper-bearing kinase in calphostin C-treated cells to facilitate apoptosis. AB - Although tissue transglutaminase (tTG) has been recognized as a mediator of apoptosis in various experimental models, little is currently known about the molecular mechanisms by which this protein modulates cell death. Recent work from our laboratory has shown that activation of tTG in cells exposed to the apoptotic inducer calphostin C triggers the crosslinking of dual leucine zipper-bearing kinase (DLK), a proapoptotic kinase acting as an essential component of the c-Jun amino-terminal kinase (JNK) signaling pathway. As a consequence of this observation, we have undertaken experiments to investigate the functional relevance of DLK oligomerization in tTG-mediated apoptosis. Our results indicate that, in cells undergoing calphostin C-induced apoptosis, tTG-dependent DLK oligomerization occurs early in the apoptotic response. Both immunocomplex kinase assays and immunoblotting with phosphospecific antibodies revealed that oligomer formation by tTG-mediated crosslinking reactions significantly enhanced the kinase activity of DLK and its ability to activate the JNK pathway. Moreover, functional studies demonstrate that tTG-mediated oligomerization of wild-type DLK sensitizes cells to calphostin C-induced apoptosis, while crosslinking of a kinase-inactive variant of DLK does not. Collectively, these data strongly suggest that tTG facilitates apoptosis, at least partly, by oligomerization and activation of the proapoptotic kinase DLK. PMID- 14739944 TI - Tell it how it is. PMID- 14739945 TI - Primary dentition. PMID- 14739946 TI - Primary dentition. PMID- 14739947 TI - Disappearing lesions. PMID- 14739957 TI - Orthodontics. Part 6: Risks in orthodontic treatment. AB - Orthodontics has the potential to cause significant damage to hard and soft tissues. The most important aspect of orthodontic care is to have an extremely high standard of oral hygiene before and during orthodontic treatment. It is also essential that any carious lesions are dealt with before any active treatment starts. Root resorption is a common complication during orthodontic treatment but there is some evidence that once appliances are removed this resorption stops. Some of the risk pointers for root resorption are summarised. Soft tissue damage includes that caused by archwires but also the more harrowing potential for headgears to cause damage to eyes. It is essential that adequate safety measures are included with this type of treatment. PMID- 14739958 TI - Necrotizing sialometaplasia: an unusual bilateral presentation associated with antecedent anaesthesia and lack of response to intralesional steroids. Case report and review of the literature. AB - Necrotizing sialometaplasia is a self-limiting, variably ulcerated benign process affecting minor salivary glands. Accurate histological diagnosis is paramount, as it has been mistaken for malignancy, which has resulted in excessively aggressive and unnecessary radical surgery. A unique case of bilateral necrotizing sialometaplasia, presenting with anaesthesia of the greater palatine nerves, is described. An attempt at active therapy with intralesional steroids had no effect on the course of the condition. PMID- 14739959 TI - Temporary mental nerve paraesthesia secondary to orthodontic treatment--a case report and review. AB - Patients complaining of facial paraesthesia may present to the dental practitioner. The differential diagnosis includes malignant neoplastic disease and therefore warrants prompt hospital referral. Mental nerve paraesthesia during orthodontic treatment is a very rare occurrence. A case of mental paraesthesia during fixed upper and lower orthodontic treatment is presented. This report highlights the importance of thorough investigation of patients presenting with a history of facial numbness, and provides a review of the aetiology and management of this problem for the dental practitioner. Guidelines for referral have been suggested where orthodontic treatment is suspected as the cause. PMID- 14739966 TI - Dental caries experience in older people over time: what can the large cohort studies tell us? AB - BACKGROUND: Little was known of the natural history of dental caries among older adults until recently, but reports from a number of large cohort studies have now enabled better understanding of the nature and determinants of dental caries in older people. The aim of this review is to examine and compare findings from established population-based longitudinal studies of older adults in order to determine their preventive implications. METHODS: The dental literature was reviewed in order to identify reports on dental caries incidence from large, population-based dental longitudinal studies of older adults (age 50+) with at least 3 years of follow-up. RESULTS: Reports were identified from four studies (in Iowa, North Carolina, Ontario and South Australia) which met the criteria; four reports dealt with coronal caries, and five with root surface caries. When annualised, coronal and root surface caries increments were combined and compared with those reported for adolescents, the caries experience of older people over time (between 0.8 and 1.2 new surfaces affected per year) exceeded that reported from cohort studies of adolescents (between 0.4 and 1.2 surfaces per year). The only caries risk factor common to all four studies was the wearing of a partial denture (for root surface caries only). CONCLUSIONS: Older people are a caries active group, experiencing new disease at a rate which is at least as great as that of adolescents. PRACTICE IMPLICATIONS: Dentate older people should be the target of intensive monitoring and preventive efforts at both the clinical practice and public health levels. There is no easily identifiable 'magic bullet' for preventing caries in that age group, but the use of evidence-based preventive interventions (such as fluoride) should suffice. PMID- 14739967 TI - A survey of the opinions of consultant anaesthetists in Scotland of sedation carried out by dentists. AB - OBJECTIVES: To elicit the attitudes and opinions of consultant anaesthetists working in Scotland, with regard to conscious sedation carried out by dental practitioners. METHOD: A questionnaire was designed to gauge opinion of consultant anaesthetists in Scotland on the practice of conscious sedation by dentists. The questionnaire was sent to 353 consultant anaesthetists working in 49 hospitals within the 15 health boards in Scotland. RESULTS: Of the 366 questionnaires sent, 249 were returned of which 235 were valid. This gave a response rate of 64%. In general, those questioned felt that the provision of sedation in a hospital setting was more appropriate than in general dental practice. A majority (65%) thought that it was unrealistic for anaesthetists to provide all sedation for dental treatment, although many (58%) felt that anaesthetists should take more responsibility in this area. Again, a majority (60%) agreed that dentists should be trained to use sedation techniques for their patients but a significant number (63%) disagreed with the practice of operator/sedationist. CONCLUSION: It is of concern to the dental profession that a significant number of anaesthetists do not feel that it is appropriate for dentists to be administering even the most simple methods of sedation. At present there are no clear, recognised guidelines as to the level of formal training required for the practice of conscious sedation by dentists. It is in the interests of the dental profession and the public to ensure that those choosing to practice sedation do so safely by following recognised guidelines in the training and practice of sedation. PMID- 14739968 TI - Effectiveness of a communication skills training programme for the management of dental anxiety. AB - The aim of this study was to develop and evaluate a communication skills training programme for the management of dental anxiety. The aims of the training were directed at the enhancement of knowledge and communication skills. The research design consisted of a pre-test-post-test-control group design. The instruments were a knowledge test, a behavioural role-play test and a learner report. Thirty four graduate students participated in the study. The results showed that the communication skills training had an effect on the knowledge and a substantial effect on the behaviour of the students. Moreover, the results from the learner report showed that the students acquired important insights in their own capacities and limitations. The conclusions are that the course as a whole is effective for dealing with anxious patients. Finally, it is recommended that knowledge and behaviour examinations are introduced as a regular part of the curricula for dentistry students. PMID- 14739985 TI - Non-homologous DNA end joining. AB - DNA double-strand breaks (DSBs) are a serious threat for the cell and when not repaired or misrepaired can result in mutations or chromosome rearrangements and eventually in cell death. Therefore, cells have evolved a number of pathways to deal with DSB including homologous recombination (HR), single-strand annealing (SSA) and non-homologous end joining (NHEJ). In mammals DSBs are primarily repaired by NHEJ and HR, while HR repair dominates in yeast, but this depends also on the phase of the cell cycle. NHEJ functions in all kinds of cells, from bacteria to man, and depends on the structure of DSB termini. In this process two DNA ends are joined directly, usually with no sequence homology, although in the case of same polarity of the single stranded overhangs in DSBs, regions of microhomology are utilized. The usage of microhomology is common in DNA end joining of physiological DSBs, such as at the coding ends in V(D)J (variable(diversity) joining) recombination. The main components of the NHEJ system in eukaryotes are the catalytic subunit of DNA protein kinase (DNA PK(cs)), which is recruited by DNA Ku protein, a heterodimer of Ku70 and Ku80, as well as XRCC4 protein and DNA ligase IV. A complex of Rad50/Mre11/Xrs2, a family of Sir proteins and probably other yet unidentified proteins can be also involved in this process. NHEJ and HR may play overlapping roles in the repair of DSBs produced in the S phase of the cell cycle or at replication forks. Aside from DNA repair, NHEJ may play a role in many different processes, including the maintenance of telomeres and integration of HIV-1 genome into a host genome, as well as the insertion of pseudogenes and repetitive sequences into the genome of mammalian cells. Inhibition of NHEJ can be exploited in cancer therapy in radio sensitizing cancer cells. Identification of all key players and fundamental mechanisms underlying NHEJ still requires further research. PMID- 14739986 TI - Effects of distortions by A-tracts of promoter B-DNA spacer region on the kinetics of open complex formation by Escherichia coli RNA polymerase. AB - A-tracts in DNA due to their structural morphology distinctly different from the canonical B-DNA form play an important role in specific recognition of bacterial upstream promoter elements by the carboxyl terminal domain of RNA polymerase alpha subunit and, in turn, in the process of transcription initiation. They are only rarely found in the spacer promoter regions separating the -35 and -10 recognition hexamers. At present, the nature of the protein-DNA contacts formed between RNA polymerase and promoter DNA in transcription initiation can only be inferred from low resolution structural data and mutational and crosslinking experiments. To probe these contacts further, we constructed derivatives of a model Pa promoter bearing in the spacer region one or two An (n = 5 or 6) tracts, in phase with the DNA helical repeat, and studied the effects of thereby induced perturbation of promoter DNA structure on the kinetics of open complex (RPo) formation in vitro by Escherichia coli RNA polymerase. We found that the overall second-order rate constant ka of RPo formation, relative to that at the control promoter, was strongly reduced by one to two orders of magnitude only when the A tracts were located in the nontemplate strand. A particularly strong 30-fold down effect on ka was exerted by nontemplate A-tracts in the -10 extended promoter region, where an involvement of nontemplate TG (-14, -15) sequence in a specific interaction with region 3 of sigma-subunit is postulated. A-tracts in the latter location caused also 3-fold slower isomerization of the first closed transcription complex into the intermediate one that precedes formation of RPo, and led to two-fold faster dissociation of the latter. All these findings are discussed in relation to recent structural and kinetic models of RPo formation. PMID- 14739987 TI - UV- and MMS-induced mutagenesis of lambdaO(am)8 phage under nonpermissive conditions for phage DNA replication. AB - Mutagenesis in Escherichia coli, a subject of many years of study is considered to be related to DNA replication. DNA lesions nonrepaired by the error-free nucleotide excision repair (NER), base excision repair (BER) and recombination repair (RR), stop replication at the fork. Reinitiation needs translesion synthesis (TLS) by DNA polymerase V (UmuC), which in the presence of accessory proteins, UmuD', RecA and ssDNA-binding protein (SSB), has an ability to bypass the lesion with high mutagenicity. This enables reinitiation and extension of DNA replication by DNA polymerase III (Pol III). We studied UV- and MMS-induced mutagenesis of lambdaO(am)8 phage in E. coli 594 sup+ host, unable to replicate the phage DNA, as a possible model for mutagenesis induced in nondividing cells (e.g. somatic cells). We show that in E. coli 594 sup+ cells UV- and MMS-induced mutagenesis of lambdaO(am)8 phage may occur. This mutagenic process requires both the UmuD' and C proteins, albeit a high level of UmuD' and low level of UmuC seem to be necessary and sufficient. We compared UV-induced mutagenesis of lambdaO(am)8 in nonpermissive (594 sup+) and permissive (C600 supE) conditions for phage DNA replication. It appeared that while the mutagenesis of lambdaO(am)8 in 594 sup+ requires the UmuD' and C proteins, which can not be replaced by other SOS-inducible protein(s), in C600 supE their functions may be replaced by other inducible protein(s), possibly DNA polymerase IV (DinB). Mutations induced under nonpermissive conditions for phage DNA replication are resistant to mismatch repair (MMR), while among those induced under permissive conditions, only about 40% are resistant. PMID- 14739988 TI - Non-random distribution of GATC sequences in regions of promoters stimulated by the SeqA protein of Escherichia coli. AB - The SeqA protein of Escherichia coli is not only the main negative regulator of DNA replication initiation but also a specific transcription factor. It binds to hemimethylated GATC sequences and, with somewhat different specificity, to fully methylated GATC regions. Recently, a microarray analysis was reported, in which transcriptomes of wild-type and DeltaseqA strains were compared. Although in the seqA mutant the levels of some transcripts were significantly decreased while certain transcripts were evidently more abundant relative to wild-type bacteria, no correlation between the presence of GATC motifs in promoter sequences and transcription activity was found. However, here we show that when larger DNA fragments, encompassing positions from -250 to +250 relative to the transcription start site, are analyzed, some common features of GATC distribution near the promoters activated by SeqA can be demonstrated. Nevertheless, it seems that the GATC pattern is not the only determinant of SeqA-dependence of promoter activity. PMID- 14739989 TI - Analogs of diadenosine tetraphosphate (Ap4A). AB - This review summarizes our knowledge of analogs and derivatives of diadenosine 5',5"'-P1,P4-tetraphosphate (Ap4A), the most extensively studied member of the dinucleoside 5',5"'-P1,Pn-polyphosphate (NpnN) family. After a short discussion of enzymes that may be responsible for the accumulation and degradation of Np4)N's in the cell, this review focuses on chemically and/or enzymatically produced analogs and their practical applications. Particular attention is paid to compounds that have aided the study of enzymes involved in the metabolism of Ap4A (Np4N'). Certain Ap4A analogs were alternative substrates of Ap4A-degrading enzymes and/or acted as enzyme inhibitors, some other helped to establish enzyme mechanisms, increased the sensitivity of certain enzyme assays or produced stable enzyme:ligand complexes for structural analysis. PMID- 14739990 TI - Degradation of extracellular nucleotides and their analogs in HeLa and HUVEC cell cultures. AB - The use of nucleotides and their analogs in the pharmacological studies of nucleotide receptors (P2 class) should be preceded by detailed studies on their degradation connected with ecto-enzymes of a given cell type. In the present studies we have analyzed stability of some phosphorothioate and phosphonate analogs of ATP and ADP in the HeLa epitheloid carcinoma and endothelial HUVEC cells cultures. Our studies have revealed that ecto-nucleotide pyrophosphatase (E NPP) is one of the main enzymes involved in the extracellular degradation of ATP and other nucleotides in the HeLa cells. On the other hand, the ecto-ATPDase is responsible for the hydrolysis of extracellular nucleotides in human endothelial cell cultures, while the E-NPP-like enzymes of the HUVEC cells are not essential to this degradation. The concerted action of the aforementioned ecto-enzymes and nucleotide pyrophosphatase, 5'-nucleotidase and adenosine deaminase present in fetal bovine serum (FBS) supplied to the culture medium, results in partial or complete degradation of the phosphorothioate (ATPgammaS) and phosphonate analogs of adenosine nucleotides (alpha,beta-methylene-ATP and beta,gamma-methylene-ATP) in the cell cultures. Only ADPbetaS appears to be resistant to these enzymes. The influence of some nucleotides and their analogs on the proliferation of the HeLa cells in presence or absence of FBS is also discussed. PMID- 14739991 TI - PDZ domains - common players in the cell signaling. AB - PDZ domains are ubiquitous protein interaction modules that play a key role in cellular signaling. Their binding specificity involves recognition of the carboxyl-terminus of various proteins, often belonging to receptor and ion channel families. PDZ domains also mediate more complicated molecular networks through PDZ-PDZ interactions, recognition of internal protein sequences or phosphatidylinositol moieties. The domains often form a tandem of multiple copies, but equally often such tandems or single PDZ domain occur in combination with other signaling domains (for example SH3, DH/PH, GUK, LIM, CaMK). Common occurrence of PDZ domains in Metazoans strongly suggests that their evolutionary appearance results from the complication of signaling mechanisms in multicellular organisms. Here, we focus on their structure, specificity and role in signaling pathways. PMID- 14739992 TI - The roles of annexins and alkaline phosphatase in mineralization process. AB - In this review the roles of specific proteins during the first step of mineralization and nucleation are discussed. Mineralization is initiated inside the extracellular organelles-matrix vesicles (MVs). MVs, containing relatively high concentrations of Ca2+ and inorganic phosphate (Pi), create an optimal environment to induce the formation of hydroxyapatite (HA). Special attention is given to two families of proteins present in MVs, annexins (AnxAs) and tissue nonspecific alkaline phosphatases (TNAPs). Both families participate in the formation of HA crystals. AnxAs are Ca2+ - and lipid-binding proteins, which are involved in Ca2+ homeostasis in bone cells and in extracellular MVs. AnxAs form calcium ion channels within the membrane of MVs. Although the mechanisms of ion channel formation by AnxAs are not well understood, evidence is provided that acidic pH or GTP contribute to this process. Furthermore, low molecular mass ligands, as vitamin A derivatives, can modulate the activity of MVs by interacting with AnxAs and affecting their expression. AnxAs and other anionic proteins are also involved in the crystal nucleation. The second family of proteins, TNAPs, is associated with Pi homeostasis, and can hydrolyse a variety of phosphate compounds. ATP is released in the extracellular matrix, where it can be hydrolyzed by TNAPs, ATP hydrolases and nucleoside triphosphate (NTP) pyrophosphohydrolases. However, TNAP is probably not responsible for ATP dependent Ca2+/phosphate complex formation. It can hydrolyse pyrophosphate (PPi), a known inhibitor of HA formation and a byproduct of NTP pyrophosphohydrolases. In this respect, antagonistic activities of TNAPs and NTP pyrophosphohydrolases can regulate the mineralization process. PMID- 14739993 TI - Prohibitins and Ras2 protein cooperate in the maintenance of mitochondrial function during yeast aging. AB - The yeast Saccharomyces cerevisiae has a finite replicative life span. Yeasts possess two prohibitins, Phb1p and Phb2p, in similarity to mammalian cells. These proteins are located in the inner mitochondrial membrane, where they are involved in the processing of newly-synthesized membrane proteins. We demonstrate that the elimination of one or both of the prohibitin genes in yeast markedly diminished the replicative life span of cells that lack fully-functional mitochondria, while having no effect on cells with functioning mitochondria. This deleterious effect was suppressed by the deletion of the RAS2 gene. The expression of PHB1 and PHB2 declined gradually up to 5-fold during the life span. Cells in which PHB1 was deleted in conjunction with the absence of a mitochondrial genome displayed remarkable changes in mitochondrial morphology, distribution, and inheritance. This loss of mitochondrial integrity was not seen in cells devoid of PHB1 but possessing an intact mitochondrial genome. In a subset of the cells, the changes in mitochondrial integrity were associated with increased production of reactive oxygen species, which co-localized with the altered mitochondria. The mitochondrial deficits described above were all suppressed by deletion of RAS2. Our data, together with published information, are interpreted to provide a unified view of the role of the prohibitins in yeast aging. This model posits that the key initiating event is a decline in mitochondrial function, which leads to progressive oxidative damage that is exacerbated in the absence of the prohibitins. This aggravation of the initial damage is ameliorated by the suppression of the production of mitochondrial proteins in the absence of Ras2p signaling of mitochondrial biogenesis. PMID- 14739994 TI - Immunodetection of surfactant proteins in human organ of Corti, Eustachian tube and kidney. AB - The presence of surfactant proteins was investigated in the human organ of Corti, Eustachian tube and kidney tissues. It has previously been shown that lamellar bodies are present in hairy cells of organ of Corti, in the cytoplasm of secretory and lumen of tubal glands of Eustachian tube and kidney renal basement membrane. No evidence for the presence of surfactant proteins in the organ of Corti and kidney has been presented until now. The aim of this study was to find out if surfactant proteins were expressed in other epithelia such as organ of Corti, Eustachian tube and kidney. Surfactant proteins were identified using one dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting. On one-dimensional Western blots, bands for surfactant protein A in human Eustachian tube (SP-A, 34 kDa) and in kidney extracts, and for surfactant protein D (SP-D, 43 kDa) in Eustachian tube and in kidney extracts (SP-D, 86 kDa), and for surfactant protein B (SP-B, 8 kDa) in human Eustachian tube and organ of Corti extracts were detected. Bands corresponded to monomeric forms of lung surfactant proteins. These results indicate the presence of SP-A and SP-D in kidney epithelium, SP-A, SP-B and SP-D in Eustachian tube and SP-B in the organ of Corti. PMID- 14739995 TI - Expression level of Ubc9 protein in rat tissues. AB - Ubc9 is a homologue of the E2 ubiquitin conjugating enzyme and participates in the covalent linking of SUMO-1 molecule to the target protein. In this report we describe a simple and efficient method for obtaining pure human recombinant Ubc9 protein. The purified Ubc9 retained its native structure and was fully active in an in vitro sumoylation assay with the promyelocytic leukaemia (PML) peptide as a substrate. In order to better understand the physiology of Ubc9 protein we examined its levels in several rat tissues. Immunoblot analyses performed on tissue extracts revealed quantitative and qualitative differences in the expression pattern of Ubc9. The Ubc9 protein was present at a high level in spleen and lung. Moderate level of Ubc9 was detected in kidney and liver. Low amount of Ubc9 was observed in brain, whereas the 18 kDa band of Ubc9 was barely visible or absent in heart and skeletal muscle. In heart and muscle extracts the Ubc9 antibodies recognized a 38 kDa protein band. This band was not visible in extracts of other rat tissues. A comparison of the relative levels of Ubc9 mRNA and protein indicated that the overall expression level of Ubc9 was the highest in spleen and lung. In spleen, lung, kidney, brain, liver and heart there was a good correlation between the 18 kDa protein and Ubc9 mRNA levels. In skeletal muscle the Ubc9 mRNA level was unproportionally high comparing to the level of the 18 kDa protein. The presented data indicate that in the rat the expression of the Ubc9 protein appears to have some degree of tissue specificity. PMID- 14739997 TI - Isozymes delta of phosphoinositide-specific phospholipase C and their role in signal transduction in the cell. AB - Phospholipase C (PLC, EC 3.1.4.11) is an enzyme crucial for the phosphoinositol pathway and whose activity is involved in eukaryotic signal transduction as it generates two second messengers: diacylglycerol (DAG) and inositol 1,4,5 trisphosphate (IP3). There are four major types of phospholipase C named: beta, gamma, delta and the recently discovered epsilon, but this review will focus only on the recent advances for the delta isozymes of PLC. So far, four delta isozymes (named delta1-4) have been discovered and examined. They differ with regard to cellular distribution, activities, biochemical features and involvement in human ailments. PMID- 14739998 TI - The effect of cAMP and cGMP on the activity and substrate specificity of protein kinase A from methylotrophic yeast Pichia pastoris. AB - Cyclic AMP dependent protein kinase (PKA) from Pichia pastoris yeast cells was found to be activated by either cAMP or cGMP. Analogs of cAMP such as 8-chloro cAMP and 8-bromo-cAMP were as potent as cAMP in PKA activation while N6,2'-O dibutyryl-cAMP did not stimulate the enzyme activity. It was shown that protamine sulfate was almost equally phosphorylated in the presence of 1-2 x 10(-6)M cAMP or cGMP while other substrates such as Kemptide, ribosomal protein S6, were phosphorylated to a lower extent in the presence of cGMP. It was demonstrated that pyruvate kinase is a substrate of PKA which co-purified with the P.pastoris enzyme. Moreover, pyruvate kinase was phosphorylated by PKA in the presence of cAMP and cGMP to comparable levels. PMID- 14739996 TI - Photoreceptor guanylate cyclase variants: cGMP production under control. AB - Changes in the Ca2+ concentration are thought to affect many processes, including signal transduction in a vast number of biological systems. However, only in few cases the molecular mechanisms by which Ca2+ mediates its action are as well understood as in phototransduction. In dark-adapted photoreceptor cells, the equilibrium level of cGMP is maintained by two opposing activities, such as phosphodiesterase (PDE) and guanylate cyclase (GC). Upon absorption of photons, rhodopsin-G-protein-mediated activation of PDE leads to a transient decrease in [cGMP] and subsequently to lowering of [Ca2+]. In turn, lower [Ca2+] increases net production of cGMP by stimulation of GC until dark conditions are re established. This activation of GC is mediated by Ca2+ -free forms of Ca2+ binding proteins termed GC-activating proteins (GCAPs). The last decade brought the molecular identification of GCs and GCAPs in the visual system. Recent efforts have been directed toward understanding the properties of GC at the physiological and structural levels. Here, we summarize the recent progress and present a list of topics of ongoing research. PMID- 14739999 TI - Inhibition of collagen-induced platelet reactivity by DGEA peptide. AB - Direct interactions between collagen, the most thrombogenic component of the extracellular matrix, and platelet surface membrane receptors mediate platelet adhesion and induce platelet activation and aggregation. In this process two glycoproteins are crucial: integrin alpha2beta1, an adhesive receptor, and GPVI, which is especially responsible for signal transduction. Specific antagonists of the collagen receptors are useful tools for investigating the complexity of platelet-collagen interactions. In this work we assessed the usefulness of DGEA peptide (Asp-Gly-Glu-Ala), the shortest collagen type I-derived motif recognised by the collagen-binding integrin alpha2beta1, as a potential antagonist of collagen receptors. We examined platelet function using several methods including platelet adhesion under static conditions, platelet function analyser PFA-100TM, whole blood electric impedance aggregometry (WBEA) and flow cytometry. We found that DGEA significantly inhibited adhesion, aggregation and release reaction of collagen activated blood platelets. The inhibitory effect of DGEA on static platelet adhesion reached sub-maximal values at millimolar inhibitor concentrations, whereas the specific blocker of alpha2beta1 - monoclonal antibodies Gi9, when used at saturating concentrations, had only a moderate inhibitory effect on platelet adhesion. Considering that 25-30% of total collagen binding to alpha2beta1 is specific, we conclude that DGEA is a strong antagonist interfering with a variety of collagen-platelet interactions, and it can be recognised not only by the primary platelet adhesion receptor alpha2beta1 but also by other collagen receptors. PMID- 14740000 TI - Melatonin as an antioxidant: biochemical mechanisms and pathophysiological implications in humans. AB - This brief resume enumerates the multiple actions of melatonin as an antioxidant. This indoleamine is produced in the vertebrate pineal gland, the retina and possibly some other organs. Additionally, however, it is found in invertebrates, bacteria, unicellular organisms as well as in plants, all of which do not have a pineal gland. Melatonin's functions as an antioxidant include: a), direct free radical scavenging, b), stimulation of antioxidative enzymes, c), increasing the efficiency of mitochondrial oxidative phosphorylation and reducing electron leakage (thereby lowering free radical generation), and 3), augmenting the efficiency of other antioxidants. There may be other functions of melatonin, yet undiscovered, which enhance its ability to protect against molecular damage by oxygen and nitrogen-based toxic reactants. Numerous in vitro and in vivo studies have documented the ability of both physiological and pharmacological concentrations to melatonin to protect against free radical destruction. Furthermore, clinical tests utilizing melatonin have proven highly successful; because of the positive outcomes of these studies, melatonin's use in disease states and processes where free radical damage is involved should be increased. PMID- 14740001 TI - Selenium, glutathione and glutathione peroxidases in blood of patients with chronic liver diseases. AB - Disturbances in the antioxidant system could play a role in pathogenesis of chronic liver disease. The aim of our study was to evaluate the levels/activities of antioxidants in blood of patients with chronic liver disease. We estimated selenium and glutathione concentrations and glutathione peroxidase activities in blood of 59 patients with chronic hepatitis B or C virus infection (group 1) and 64 patients with alcoholic, autoimmune or cryptogenic chronic liver disease (group 2). The results were compared with 50 healthy controls. Whole blood and plasma selenium and red cell glutathione concentrations were significantly lower in the patients compared with the controls. Red cell glutathione peroxidase activity was slightly reduced in both subgroups of group 1 and in group 2 with normal alanine aminotransferase values. Plasma glutathione peroxidase activity was slightly but significantly higher in patients with elevated aminotransferase values. The findings suggest that disturbances in antioxidant parameters in blood of patients with chronic liver disease may be the cause of the peroxidative damage of cells. PMID- 14740002 TI - Application of electrophoretic methods for detection of protein-porphyrin complexes. AB - Simple methods for detection and isolation of protein-porphyrin complexes were elaborated in our laboratory. They are based on the separation of protein porphyrin complexes in native polyacrylamide gel and measurement of their fluorescence, with the use of two detection systems: the commercially available Gel Doc(TM) 2000 system, and a system specially designed for the purpose of these investigations, concerning protein-porphyrin interactions. The fluorescent complexes can be electro-transferred from the gel onto PVDF membrane, eluted and analyzed in order to identify the protein interacting with porphyrins. PMID- 14740003 TI - Photodynamic effects of two water soluble porphyrins evaluated on human malignant melanoma cells in vitro. AB - Two water soluble porphyrins: meso-tetra-4-N-methylpyridyl-porphyrin iodide (P1) and 5,10-di-(4-acetamidophenyl)-15,20-di-(4-N-methylpyridyl) porphyrin (P2) were synthesised and evaluated in respect to their photochemical and photophysical properties as well as biological activity. Cytotoxic and phototoxic effects were evaluated in human malignant melanoma Me45 line using clonogenic assay, cytological study of micronuclei, apoptosis and necrosis frequency and inhibition of growth of megacolonies. Both porphyrins were characterised by high UV and low visible light absorptions. Dark toxicity measured on the basis of the clonogenic assay and inhibition of megacolony growth area indicated that P1 was non-toxic at concentrations up to 50 microg/ml (42.14 microM) and P2 at concentrations up to 20 microg/ml (16.86 microM). The photodynamic effect induced by red light above 630 nm indicated that both porphyrins were able to inhibit growth of melanoma megacolonies at non-toxic concentrations. Cytologic examination showed that the predominant mode of cell death was necrosis. PMID- 14740004 TI - A study of the influence of newly synthesized acyclonucleosides and 1,2,3,4 tetrahydroisoquinoline derivatives on deoxythymidine and deoxycytidine kinase activities in human neurofibrosarcoma and ovarian cancer. AB - The influence of nine newly synthesized uracil acyclonucleosides, and 36 derivatives of 1,2,3,4-tetrahydroisoquinoline on the activity of enzymes catalysing dTMP and dGMP synthesis, on the content of dTTP and dGTP in acid soluble fraction and on the incorporation of [14C]dThd and [14C ]dGuo into DNA in tumour homogenates was studied. The influence of the compounds was studied in the cytosol from intraoperatively excised human tumours - neurofibrosarcoma and ovarian cancer. It was shown that dTMP and dGMP synthesis is inhibited competitively by 34.1+/-4.0% in both types of tumours by 0.2 mM 1-N-(3' hydroxypropyl)-6-methyluracil (1) and 0.2 mM 1-N-(3'-hydroxypropyl)- 5,6- tetramethyleneuracil (2). The mentioned acyclonucleosides reduced the content of dTTP and dGTP in the acid soluble fraction of tumours (59.7+/-3.1% of control). 1 (4-chlorophenyl)-6,7-dihydroxy- 1,2,3,4-tetrahydroisoquinoline (3), 1-(2,3 dichlorophenyl)-6,7-dihydroxy 1,2,3,4-tetrahydroisoquinoline (4) and 1-(3 methoxyphenyl)-6,7-dihydroxy 1,2,3,4-tetrahydroisoquinoline (5) at 0.2 mM concentration caused a mixed type inhibition of the synthesis of dTMP and dGMP by, on average, 33.2+/-4.4%, and reduced the content of dTTP and dGTP in the acid soluble fraction (52.6+/-3.7% of control) but were active only in the cytosol of neurofibrosarcoma. While acyclonucleosides undergo phosphorylation in the cytosol by cellular kinases, with their triphosphates being active acyclonucleoside metabolites, active 1,3,4,5-tetrahydroisoquinoline derivatives (compounds not containing a deoxyribose moiety), cannot be phosphorylated. ACN and THI derivatives which inhibit dThd and dCyd kinase activities, inhibit also the incorporation of [14C]dThd and [14C]dGuo (ACN - 50.2+/-2.7%, THI - 53.4+/-3.9% of incorporation inhibition) into tumour DNA. The obtained results point to the mechanism of uracil acyclonucleosides and 1,2,3,4-tetrahydroisoquinoline biological activity consisting in inhibiting the synthesis of DNA components. PMID- 14740005 TI - Comparative analysis of RCAS1 level in neoplasms and placenta. AB - The tumor associated antigen RCAS1 (receptor-binding cancer antigen expressed on SiSo cells) expressed with high frequency in various cancer and trophoblast cells, inhibits growth of estrogen receptor-expressing cells and induces apoptosis. Because previous reports demonstrated RCAS1 presence only by non quantitative immunocytochemistry methods, we decided to use a Western blotting with anti-RCAS1 monoclonal antibodies for estimation of the relative content of the tumor-associated antigen. One hundred tissue samples were assayed (neoplasms, chronic inflammatory diseases, healthy tissues, trophoblasts and placentas at term). RCAS1 was present in all neoplastic, placental and trophoblast tissue samples and its level in malignant samples was statistically significantly higher than in benign neoplasms. The amount of RCAS1 in chronic inflammations was also significantly increased in immune mediated diseases, like allergic nasal polyps and sarcoidosis. The RCAS1 protein was not revealed in healthy mucous membrane and in muscle tissues. The presented results suggest that RCAS1 might play an important role in tumor escape from host immunological surveillance and carry weight in the down regulation of the maternal immune response, thereby maintaining pregnancy. PMID- 14740006 TI - Analysis of microsatellite instability and loss of heterozygosity in breast cancer with the use of a well characterized multiplex system. AB - Analysis of microsatellite instability (MI) and loss of heterozygosity (LOH) is recommended for screening patients with sporadic and hereditary malignancies. This study shows an application of a fluorescent hexaplex PCR system for microsatellite typing on A.L.F. DNA Sequencer (Pharmacia Biotech). This technique detects changes in microsatellites providing a time-efficient, reliable and accurate method for MI and LOH analyses. The Fragment Manager software was used for automated size calculation and quantitation of DNA fragments, enabling rapid and precise measurement of allelic ratios. We examined 70 breast cancer and 70 control DNA specimens, classified all the patterns of microsatellite alterations, and set up MI and LOH assessment criteria for the automated multiplex fluorescent method. PMID- 14740007 TI - Paul-Bunnell antigen and a possible mechanism of formation of heterophile antibodies in patients with infectious mononucleosis. AB - Sera of patients with infectious mononucleosis contain heterophile anti-Paul- Bunnell (PB) antibodies to erythrocytes of numerous mammalian species. Evidence is presented that the corresponding antigen of bovine erythrocytes is not, as previously described, a single molecule, but a series of glycoproteins with glycans terminated with N-glycolylneuraminic acid (Neu5Gc). The latter compound should be an important part of the PB epitope because, in agreement with the results of others, we found that desialylation of the PB antigen abolishes almost completely its activity. We examined three different preparations of GM3 ganglioside for their capacity to bind anti-PB and found that only GM3 from horse erythrocytes containing Neu5Gc exhibited a low although ELISA measurable PB activity. The other two GM3 preparations, from bovine milk and dog erythrocytes, containing N-acetylneuraminic acid (Neu5Ac) bound little if any anti-PB antibodies. This finding confirms a previous report that human erythrocyte Neu5Ac containing sialoglycoprotein with similar O-linked glycans as the PB-antigen of bovine erythrocytes exhibits only very low PB activity (Patarca & Fletcher, 1995, Crit Rev Oncogen., 6: 305). In conclusion, we present a hypothesis that anti-PB antibodies in patients with infectious mononucleosis are formed against infection induced cell membrane glycoconjugates containing highly immunogenic Neu5Gc. PMID- 14740008 TI - The structure and protein binding of amyloid-specific dye reagents. AB - The self-assembling tendency and protein complexation capability of dyes related to Congo red and also some dyes of different structure were compared to explain the mechanism of Congo red binding and the reason for its specific affinity for beta-structure. Complexation with proteins was measured directly and expressed as the number of dye molecules bound to heat-aggregated IgG and to two light chains with different structural stability. Binding of dyes to rabbit antibodies was measured indirectly as the enhancement effect of the dye on immune complex formation. Self-assembling was tested using dynamic light scattering to measure the size of the supramolecular assemblies. In general the results show that the supramolecular form of a dye is the main factor determining its complexation capability. Dyes that in their compact supramolecular organization are ribbon shaped may adhere to polypeptides of beta-conformation due to the architectural compatibility in this unique structural form. The optimal fit in complexation seems to depend on two contradictory factors involving, on the one hand, the compactness of the non-covalently stabilized supramolecular ligand, and the dynamic character producing its plasticity on the other. As a result, the highest protein binding capability is shown by dyes with a moderate self-assembling tendency, while those arranging into either very rigid or very unstable supramolecular entities are less able to bind. PMID- 14740009 TI - Neural impacts on the regeneration of skeletal muscles. AB - The regeneration of skeletal muscles is a suitable model to study the development and differentiation of contractile tissues. Neural effects are one of the key factors in the regulation of this process. In the present work, effects of different reinnervation protocols (suture or grafting) were studied upon the regenerative capacity of rat soleus muscles treated with the venom of the Australian tiger snake, notexin, which is known to induce complete necrosis and subsequent regeneration of muscles. Morphological and motor endplate analysis indicated that the regenerative capacity of denervated, and thereafter surgically reinnervated muscles remains impaired compared to that of normally innervated muscles, showing differences in the muscle size, fiber type pattern and motor endplate structure, even 35 days after the notexin injection. A lack or deficiency of secreted neural factors, deterioration of satellite cells and/or incomplete recovery of the sutured or grafted nerves may be the cause of these discrepancies in the regeneration process. PMID- 14740010 TI - Evidence for the presence of the Kennedy and Bremer- Greenberg pathways in Caenorhabditis elegans. AB - Nematodes were found to synthesize phosphorylcholine-containing molecules not present in higher organisms, i.e. phosphorylcholine-substituted glycosphingolipids and (glyco)proteins. Investigations on the biosynthesis of these structures provided first biochemical evidence for the presence of the Kennedy and Bremer-Greenberg pathways in the model organism Caenorhabditis elegans. PMID- 14740011 TI - Plant purple acid phosphatases - genes, structures and biological function. AB - The properties of plant purple acid phosphatases (PAPs), metallophosphoesterases present in some bacteria, plants and animals are reviewed. All members of this group contain a characteristic set of seven amino-acid residues involved in metal ligation. Animal PAPs contain a binuclear metallic center composed of two irons, whereas in plant PAPs one iron ion is joined by zinc or manganese ion. Among plant PAPs two groups can be distinguished: small PAPs, monomeric proteins with molecular mass around 35 kDa, structurally close to mammalian PAPs, and large PAPs, homodimeric proteins with a single polypeptide of about 55 kDa. Large plant PAPs exhibit two types of structural organization. One type comprises enzymes with subunits bound by a disulfide bridge formed by cysteines located in the C terminal region around position 350. In the second type no cysteines are located in this position and no disulfide bridges are formed between subunits. Differences in structural organisation are reflected in substrate preferences. Recent data reveal in plants the occurrence of metallophosphoesterases structurally different from small or large PAPs but with metal-ligating sequences characteristic for PAPs and expressing pronounced specificity towards phytate or diphosphate nucleosides and inorganic pyrophosphate. PMID- 14740012 TI - Alternative oxidase in higher plants. AB - Plant respiratory chain branches at the level of ubiquinone from where the electrons flow through the cytochrome pathway or to alternative oxidase. Transfer of electrons from ubiquinone to oxygen by alternative oxidase has a non protonmotive character and, by bypassing two sites of H+ pumping in complexes III and IV, lowers the energy efficiency of respiration. In this paper we review theoretical and experimental studies about the structure and possible function of alternative oxidase. The evidence for specific gene expression dependent on the physiological, developmental and environmental conditions is also described. We underline the physiological role of alternative oxidase as a "survival" protein that allows plants to cope with the stressful environment. PMID- 14740013 TI - Characterisation of Mesorhizobium huakuii cyclic beta-glucan. AB - Periplasmic and extracellular glucans of Mesorhizobium huakuii were isolated and characterized by compositional and MALDI-TOF analyses, as well as 1H and 13C NMR spectroscopy. It was shown that M. huakuii produces a cyclic beta-glucan composed entirely of nonbranched glucose chains and unmodified by nonsugar substituents. The degree of polymerisation of the cyclic oligosaccharides was estimated to be in the range from 17 to 28. The most abundant glucan molecules contained 22 glucose residues. Glucose residues within the glucan were connected by beta-(1,2) glycosidic linkages. The cyclic glucan produced by M. huakuii is quite similar to the periplasmic beta-(1,2) glucans synthesized by Agrobacterium and Sinorhizobium genera. The synthesis of beta-glucan in M. huakuii is osmoregulated and this glucan could function as an osmoprotectant in free living cells. PMID- 14740014 TI - Paris street names. PMID- 14740015 TI - Art and biography. PMID- 14740017 TI - A waning comet: John Sheldon, FRS (1752-1808). AB - John Sheldon, an eccentric surgeon-anatomist, of considerable brilliance, was the first Englishman to ascend in a balloon. He became insane in 1787, after which he moved from London. He re-established himself at Exeter and was elected surgeon to the hospital in 1797. PMID- 14740018 TI - Plaques on London houses of medico-historical interest. Sir Alexander Fleming (1881-1955). PMID- 14740019 TI - Observations on Continental hospitals and practice as described by the English surgeon James Fitzjames Fraser West (1833-1883). AB - During his travels to Continental Europe, the Birmingham surgeon James Fitzjames Fraser West (1833-1883) observed innovative procedures and sound hospital practice. He also witnessed things that dismayed and sometimes disgusted him. West was renowned for recording these experiences in his diaries and sharing them with colleagues in medical society meetings and published reports. This paper provides an insight into some of these observations. PMID- 14740020 TI - Hugh Cairns (1896-1952) and the mobile neurosurgical units of World War II. AB - In World War II, Hugh Cairns, Oxford Nuffield Professor of Surgery and brigadier in the Royal Army Medical Corps, designed and administered the mobile neurosurgical units that treated casualties with head injuries in the various campaigns fought by the British Army. Cairns also created the Combined Services Hospital for Head Injuries at St Hugh's College, Oxford, where the staff of the units were trained and where evacuated casualties were received. The excellent outcome of the head-injured in World War II and the impetus to the expansion of neurosurgery in the UK during and after that war were, in large measure, due to Cairns. Others had knowledge of neurosurgery but Cairns inspired surgeons, neurologists and nursing sisters to perform neurosurgery at the highest level on the battlefield. PMID- 14740021 TI - Thomas Hodgkin's journey through Spain in 1863 and 1864. AB - In 1863 and 1864, Thomas Hodgkin travelled to Morocco, accompanying his close friend Sir Moses Montefiore, who stopped in Madrid on the way there and back to attend an audience with the Queen of Spain. Thomas Hodgkin's impressions of Spain were recounted in his book Narrative of a Journey to Morocco. The book reveals new aspects of Hodgkin's personality as well as his capacity for observation. It also describes Spain in the second half of the nineteenth century. Thus Thomas Hodgkin was a witness to one of the most vivid periods of the history of the country. The paper considers both aspects of Hodgkin's commentaries on Spain. PMID- 14740022 TI - Sir David Monro, MD (1813-1877), of New Zealand: his antecedents and his descendants. AB - David Monro, MD, was a son of the third Professor Monro of Edinburgh. He went into practice in Edinburgh and then emigrated to the Nelson settlement in New Zealand. There he practised a little medicine, but was mostly a gentleman sheep farmer. He was a member of the first parliament of New Zealand in 1854 and served as the second Speaker, when he did much to set up proper standing orders for the colonial legislature. He was knighted in 1866. PMID- 14740023 TI - Some presidents of the Medical Society of London. AB - John Coakley Lettsom (1744-1815) helped found the Medical Society of London in 1773. Its headquarters are in an elegant Regency building in central London, which contains a medical historical treasure trove. It celebrated its three hundredth session of lectures and social functions in 2003. Since 1850 the Presidency has been for one year only. In 1971 a group photograph of Presidents was arranged and a brief account of each of these distinguished men is given in the order of his seniority in the office. PMID- 14740024 TI - Medical statues, Erastus B Wolcott (1804-1880). PMID- 14740025 TI - Roman emperors suffering from apoplexy: the medical and historical significance of classical literary sources. AB - According to various Greek and Latin texts, several Roman emperors died of "apoplexy". This paper presents a systematic collection and evaluation of these sources. The contents of the texts are compared with contemporary knowledge as well as present-day perspectives. In retrospect, few of the "royal cases" can be classified as cerebrovascular disorders. PMID- 14740026 TI - Medical street names in Paris. AB - In Paris, the blue enamel signs for streets named after a person also bear the dates of birth and death, and a word or two to designate the person's field of activity. In this paper an alphabetical listing is given of 166 Parisian streets that have been named after medically qualified men who achieved distinction in medicine or elsewhere. PMID- 14740027 TI - Medical memorials. Sir Charles Hastings (1794-1866). PMID- 14740028 TI - Dignity. PMID- 14740029 TI - The tethered Biginelli condensation in natural product synthesis. AB - This review describes the development of the tethered Biginelli condensation and its application to the total synthesis of structurally complex, bioactive guanidine alkaloids. PMID- 14740030 TI - Continuous laminar evaporation: micron-scale distillation. PMID- 14740031 TI - Fluorescence properties of 2-aminopurine-cytidine-7-deazaguanine (5'-ApCdzG-3') trimer in B- and Z-DNA. AB - The electron transfer quenching of 2-aminopurine by guanine and 7-deazaguanine was investigated in B- and Z-DNA, and an increase in the fluorescence intensity of 2-aminopurine upon B- to Z-DNA transition was demonstrated. PMID- 14740032 TI - Self-assembled metal-organic squares derived from linear templates as exemplified by a polydentate ligand that provides access to both a polygon and polyhedron. PMID- 14740033 TI - Spin labeling monitors weak host-guest interactions. AB - A resorcinarene bearing four TEMPO units recognizes small molecules in solutions. PMID- 14740034 TI - Thioglycosynthases: double mutant glycosidases that serve as scaffolds for thioglycoside synthesis. AB - A double mutant, retaining glycosidase that lacks both the catalytic nucleophile and the catalytic acid/base residues efficiently catalyzes thioglycoside formation from a glycosyl fluoride donor and thiosugar acceptors. PMID- 14740035 TI - Room-temperature one-step immobilization of rod-like helical polymer onto hydrophilic substrates. PMID- 14740036 TI - Polymorphism in butylated hydroxy anisole (BHA). PMID- 14740037 TI - In flagrante metallo-cyclophane self-assembly? PMID- 14740038 TI - Parallel nucleic acid recognition by the LNA (locked nucleic acid) stereoisomers beta-L-LNA and alpha-D-LNA; studies in the mirror image world. AB - Two LNA (locked nucleic acid) stereoisomers (beta-L-LNA and alpha-D-LNA) are evaluated in the mirror-image world, that is by the study of two mixed sequences of LNA and alpha-L-LNA and their L-DNA and L-RNA complements. Both are found to display high-affinity RNA-recognition by the formation of duplexes with parallel strand orientation. PMID- 14740039 TI - Artificial muscle: movement and position control. AB - The movement of an all polymeric triple-layer artificial muscle based on polypyrrole (PPy) that work in aqueous solution was characterised. The magnitude and direction of the experimental current control both rate and direction of the movement. The described angle is a linear function of the consumed charge. PMID- 14740040 TI - Oxygenation of thiolates to S-bonded sulfinate in an iron III complex related to nitrile hydratase. AB - Air oxidation of the iron(III) complex derived from (2-mercaptoisobutyryl)-4,5 dichloro-O-phenylenediamine in the presence of Et4NCN afforded the corresponding disulfinato species. With two carboxamido nitrogens trans to two S-bonded sulfinates, this complex mimics the disulfinate inactive form of Fe-NHase. PMID- 14740041 TI - Alteration of room temperature phosphorescence lifetimes of quinine and quinidine by chiral additives. PMID- 14740042 TI - A new approach to stereospecific synthesis of P-chiral phosphorothioates. Preparation of diastereomeric dithymidyl-(3'-5') phosphorothioates. AB - A new method for stereospecific synthesis of P-chiral phosphorothioates based on intramolecular nucleophile catalysis was developed. PMID- 14740043 TI - A versatile and concise route to carbocycles using a 1,6-electrocyclic reaction. PMID- 14740044 TI - Oxidative coupling of epigallocatechin gallate amplifies antioxidant activity and inhibits xanthine oxidase activity. AB - Oxidative coupling of epigallocatechin gallate resulted in great improvement in antioxidant activity such as radical scavenging activity against superoxide anion and in activity to inhibit xanthine oxidase, offering high potential as a therapeutic agent for prevention of xanthine oxidase-induced diseases such as gout. PMID- 14740045 TI - Scintillation proximity assays for the real-time detection and quantification of the progress of reactions upon solid supports. PMID- 14740046 TI - Glyoxal bis-hydrazones: a new family of nitrogen ligands for asymmetric catalysis. PMID- 14740047 TI - Microwave-accelerated synthesis of lengthy and defect-free poly(m phenyleneethynylene)s via AB' and A2 + BB' polycondensation routes. PMID- 14740048 TI - Towards conformationally-locked difluorosugar analogues: an unexpected sense of dihydroxylation. AB - Difluorinated cyclooctenones, synthesised using RCM, can be used as templates for stereoselective oxidative transformations to products that undergo transannular reactions to afford conformationally-locked analogues of 2-deoxy-2,2 difluorosugars with different stereochemical relationships between the C-2 and C 3 hydroxyl groups. PMID- 14740049 TI - Stereoselective [2,3] sigmatropic rearrangement of acyclic allylic phosphinites. PMID- 14740050 TI - Anti-viral cyclam macrocycles : rapid zinc uptake at physiological pH. AB - NMR and UV-vis spectroscopic studies show that Zn(II) binds to cyclam rapidly at micromolar concentrations, an observation relevant to the anti-viral activity and co-receptor binding of anti-HIV cyclams. PMID- 14740051 TI - A readily-prepared electrocatalytic coating that is more active than platinum for hydrogen generation in 1 M strong acid. PMID- 14740052 TI - A powerful novel strategy for the preparation of discrete inorganic carbon-free rings containing alkaline cations. PMID- 14740053 TI - An inorganic iron complex that inhibits wild-type and an isoniazid-resistant mutant 2-trans-enoyl-ACP (CoA) reductase from Mycobacterium tuberculosis. AB - The in vitro kinetics of inactivation of both wild-type and I21V InhA enzymes by [FeII(CN)5(INH)]3- indicate that this process requires no activation by KatG, and no need for the presence of NADH. This inorganic complex may represent a new class of lead compounds to the development of anti-tubercular agents aiming at inhibition of a validated target. PMID- 14740054 TI - An FeIII wheel with a zwitterionic ligand: the structure and magnetic properties of [Fe(OMe)2(proline)]12[ClO4]12. AB - The structure and magnetic properties of a dodecanuclear iron(III) wheel are reported, featuring a bridging zwitterionic ligand. PMID- 14740055 TI - Fast, long-range electron-transfer reactions of a "blue" copper protein coupled non-covalently to an electrode through a stilbenyl thiolate monolayer. AB - A self-assembled monolayer (SAM), formed by the insitu saponification of a stilbenyl thioacetate on a gold electrode, yields fast electron transfer (ET)(the exchange rate at zero driving force exceeds 1600 s-1) with adsorbed molecules of the blue copper protein, azurin, over a distance exceeding 15 angstroms . PMID- 14740056 TI - New supramolecular polymers containing both terpyridine metal complexes and quadruple hydrogen bonding units. PMID- 14740057 TI - Photocontrol of the adsorption behavior of phenol for an azobenzene montmorillonite intercalation compound. PMID- 14740058 TI - Contrasting coordination behaviour of 22-membered chalcogenaaza (Se, Te) macrocylces towards Pd(II) and Pt(II): isolation and structural characterization of the first metallamacrocyle with a C-Pt-Se linkage. PMID- 14740059 TI - A new biotinylated tris bipyridinyl iron(II) complex as redox biotin-bridge for the construction of supramolecular biosensing architectures. AB - The bioaffine immobilization of several avidin layers on an electrode modified by a biotinylated polymer was accomplished by the first biotinylated redox bridge consisted of a tris(bipyridyl)iron(II) complex bearing six pre-oriented biotin groups. PMID- 14740060 TI - Discovery of two high-energy N2O2 isomers. PMID- 14740061 TI - Polyhedral monocarbaborane chemistry. Carboxylic acid derivatives of the [closo-2 CB9H10]- anion. PMID- 14740062 TI - Stereoselective ring-opening polymerization of racemic lactide using alkoxy-amino bis(phenolate) group 3 metal complexes. PMID- 14740063 TI - Definitive identification of lead(II)-amino acid adducts and the solid state structure of a lead-valine complex. AB - Electrospray ionization mass spectra of lead(II) nitrate-amino acid mixtures enable unequivocal identification of lead complexes for each of the essential amino acids and a valine complex is reported as the first crystallographically characterized lead-amino acid complex. PMID- 14740064 TI - Indium sulfide nanorods from single-source precursor. PMID- 14740065 TI - Partially stripped insulated nanowires: a lightly substituted hexa-peri hexabenzocoronene-based columnar liquid crystal. PMID- 14740066 TI - Polysubstituted phthalocyanines by nucleophilic substitution reactions on hexadecafluorophthalocyanines. PMID- 14740067 TI - Excited state properties of monomeric and dimeric axially bridged indium phthalocyanines upon UV-Vis laser irradiation. PMID- 14740068 TI - A chemical route from PTFE to amorphous carbon nanospheres in supercritical water. PMID- 14740069 TI - Formation of polypyrrole from 2,5-bis(2-pyrrolyl)pyrrolidine. PMID- 14740070 TI - Unexpected cleavage of tetrahydrofuran by catalytic reductive lithiation. PMID- 14740071 TI - Aligned carbon nanotube-DNA electrochemical sensors. AB - Single-strand DNA chains were chemically grafted onto aligned carbon nanotube electrodes, leading to novel aligned carbon nanotube-DNA sensors of a high sensitivity and selectivity for probing complementary DNA and target DNA chains of specific sequences. PMID- 14740072 TI - Preparation of thermosensitive PNIPAAm hydrogels with superfast response. PMID- 14740073 TI - Survival advantage with cardiovascular drugs: are they real? PMID- 14740074 TI - Atherosclerosis in type 2 diabetes mellitus: the role of insulin resistance. AB - Type 2 diabetes mellitus is associated with a marked increase in the risk of atherosclerotic diseases, including coronary heart disease, peripheral arterial disease, and cerebrovascular disease. Insulin resistance is a key factor in the pathogenesis of type 2 diabetes mellitus. Insulin resistance and its attendant metabolic abnormalities may cause much of the increased cardiovascular risk of type 2 diabetes mellitus. Among the abnormalities associated with insulin resistance are dyslipidemia, hypertension, systemic inflammation, and a prothrombotic state. This review discusses the role that each of these disorders plays in the cardiovascular risk of type 2 diabetes mellitus. PMID- 14740075 TI - Elevated levels of plasma homocysteine in hypertensive patients with diabetes mellitus. AB - BACKGROUND: Homocysteine generates oxygen radicals (superoxide anion and hydrogen peroxide) that are known to produce vasoconstriction. Hypertension is a common problem in individuals with diabetes mellitus. It is possible that hypertension in diabetic patients may be due to increased levels of plasma homocysteine. We investigated the plasma levels of homocysteine, factors involved in homocysteine metabolism (serum folic acid and vitamin B12) and lipid peroxidation product in the serum of diabetic patients with hypertension. METHODS AND RESULTS: The studies were conducted in three groups: 1) healthy controls, and diabetic patients who were 2) normotensive and 3) hypertensive. Plasma homocysteine, serum malondialdehyde (a lipid peroxidation product), vitamin B12, and folic acid were measured in these patients. Plasma homocysteine and serum malondialdehyde levels were elevated in diabetic patients compared to the control group. Plasma levels of homocysteine and serum levels of malondialdehyde were higher in the hypertensive diabetic patients than in those who were normotensive. Levels of serum folate were lower in hypertensive diabetic patients compared to the normotensive group. Levels of serum vitamin B12 were similar in both the normotensive and hypertensive diabetic patients. CONCLUSIONS: Levels of plasma homocysteine and serum malondialdehyde are elevated in hypertensive diabetic patients. Hyperhomocysteinemia may be involved in the induction and sustaining of hypertension in diabetic patients. PMID- 14740076 TI - Acid-base and electrolyte abnormalities observed in patients receiving cardiovascular drugs. AB - Cardiovascular drugs can cause a variety of acid-base and electrolyte abnormalities that need to be considered when clinicians manage the large number of patients who receive these agents. Diuretic-induced metabolic alkalosis is the most common acid-base disorder observed and is associated with hypokalemia. Drug induced hyperkalemia is the most important cause of increased potassium levels in everyday clinical practice. Multifactorial-origin diuretic-induced hyponatremia is mostly due to thiazides and should be carefully managed. This review focuses on the pathogenetic mechanisms as well as on the treatment of these metabolic derangements that are commonly encountered in patients who receive cardiovascular drugs. PMID- 14740077 TI - Effects of acetaminophen on myocardial infarct size in rats. AB - OBJECTIVE: Acetaminophen is widely prescribed as an analgesic agent in hospitals and clinics. However, acetaminophen theoretically could influence myocardial infarct size by reducing prostaglandin synthesis in vivo. To date, the effect of acetaminophen on myocardial infarct size is unknown. The present study investigated (1) whether acetaminophen has any effect on myocardial infarct size when given in an analgesic dose and (2) whether acetaminophen can affect the cardioprotective effect of the early phase of ischemic preconditioning in rats. METHODS: Female Sprague-Dawley rats were randomly assigned to four groups (n=12 each). Group 1 (no preconditioning): Vehicle (intravenous ethanol, 0.9 mL/kg) was given 39 minutes prior to ischemia. Group 2 (acetaminophen plus no preconditioning): intravenous acetaminophen (125 mg/kg) was given 39 minutes prior to ischemia. Group 3 (preconditioning): The heart was preconditioned before ischemia, and the vehicle (intravenous ethanol, 0.9 mL/kg) was given 39 minutes prior to the ischemia. Group 4 (acetaminophen plus preconditioning): The heart was preconditioned before ischemia, and intravenous acetaminophen (125 mg/kg) was given 39 minutes prior to the ischemia. The preconditioning protocol consisted of three cycles of 3 minutes of coronary occlusion and 5 minutes of reperfusion. The left coronary artery was then occluded for 60 minutes, followed by 3 hours of reperfusion. The end points were hemodynamics, body temperature, and risk area and area of necrosis of the left ventricle. RESULTS: The area of risk was similar among the four groups. The area of necrosis, expressed as a percentage of the area at risk, was 55.7% +/- 6.1% in the no-preconditioning group, 62.8% +/- 2.4% in the acetaminophen plus no-preconditioning group, 24.7% +/- 7.3% in the preconditioning group, and 17.2% +/- 6.4% in the acetaminophen plus preconditioning group. The area of necrosis/area at risk was decreased significantly in the preconditioning group and in the acetaminophen plus preconditioning group compared with the no-preconditioning group (P<.05); but there were no significant differences between the no-preconditioning group and the acetaminophen plus no-preconditioning group (P=.29), or between the preconditioning group and acetaminophen plus preconditioning group (P=.45). Among the four groups, heart rate and body temperature were similar. The infusion of the vehicle or acetaminophen increased blood pressure in the four groups, but to a lesser extent in the acetaminophen group. However, during coronary artery occlusion and reperfusion, the four groups had comparable blood pressures. CONCLUSION: Acetaminophen had no beneficial or adverse effects on infarct size in nonpreconditioned rats, and the beneficial effects of preconditioning were not blocked or prevented by acetaminophen at this analgesic dose. PMID- 14740078 TI - Effect of angiotensin II type 2 receptor blockade on activation of mitogen activated protein kinases after ischemia-reperfusion in isolated working rat hearts. AB - BACKGROUND: The stress-responsive mitogen-activated protein kinases (MAPKs) (p38 MAPK, c-Jun NH2-terminal kinase [JNK-1 and JNK-2], and extracellular signal regulated kinases [ERK-1 and ERK-2]) might be involved in angiotensin II (AII) induced ischemia-reperfusion injury. Cardioprotection induced by AII type 1 (AT1) and type 2 (AT2) receptor blockade during ischemia-reperfusion is associated with protein kinase Cepsilon (PKCepsilon), nitric oxide, and cyclic guanosine monophosphate (cGMP) signaling. Our aim was to assess the effect of selective AT1 and AT2 receptor blockade with losartan and PD123,319, respectively, on MAPK expression after ischemia-reperfusion in isolated working rat hearts. METHODS: Groups of six hearts were subjected to global ischemia (30 minutes) followed by reperfusion (30 minutes) and exposed to no drug/no ischemia-reperfusion (control), ischemia-reperfusion/no drug, and ischemia-reperfusion with losartan (1 microM), or PD123,319 (0.3 microM) and additional groups. AT1/AT2 receptor expression, MAPKs, PKCepsilon, and cGMP, and changes in mechanical function were measured. Western blotting was done on left ventricular tissue for AT1/AT2, p38/phosphorylated-p38 (p-p38), phosphorylated (p)-JNK-1/-2, phosphorylated (p) ERK-1/-2, and PKCepsilon proteins; Northern blots for AT1/AT2 mRNA; and enzyme immunoassay for cGMP. RESULTS: Compared with controls, ischemia-reperfusion induced significant left ventricular dysfunction, decreased AT2 protein and mRNA, increased p-p38 and p-JNK-1/-2, did not change p-ERK-1/-2 or PKCepsilon, and decreased cGMP. PD123,319 improved left ventricular recovery after ischemia reperfusion, increased AT2 protein and mRNA, mildly increased p-p38, normalized p JNK-1, did not change p-ERK-1/-2, and increased PKCepsilon and cGMP. Losartan did not change p-p38, increased p-JNK-1, and did not change pERK-1/-2, PKCepsilon, or cGMP. CONCLUSIONS: The overall results suggest that the activation of p38-MAPK and JNK might be linked to AII signaling and play a significant role in acute ischemia-reperfusion injury as well as in the cardioprotective effect of AT2 receptor blockade. PMID- 14740079 TI - Differential effects of peroxisome proliferator activator receptor-alpha and gamma ligands on intimal hyperplasia after balloon catheter-induced vascular injury in Zucker rats. AB - BACKGROUND: Patients with type 2 diabetes mellitus have a higher rate of restenosis following angioplasty. Peroxisome proliferator activator receptor alpha (PPAR) and gamma ligands such as fenofibrate and rosiglitazone, respectively, have been shown to have protective effects on the vessel wall. We studied the effect of fenofibrate and rosiglitazone on intimal hyperplasia in the Zucker rat, a model for insulin resistance and type 2 diabetes mellitus, following balloon catheter-induced injury. METHODS AND RESULTS: Three groups of 13-week-old female fatty Zucker rats were administered an aqueous suspension of either 3 mg/kg/d rosiglitazone (n=7) or 150 mg/kg/d fenofibrate (n=6) by gavage, or served as controls (n=9). In addition, two groups of 13-week-old female lean Zucker rats were either administered 3 mg/kg/d rosiglitazone (n=6) or served as controls (n=6). Carotid balloon injury was induced 1 week after the drugs were started. The drug administration was continued for 3 weeks. A 2-mm balloon catheter was introduced through the femoral artery to the left carotid. The balloon was inflated to 4 atmospheres for 20 seconds and then was deflated to 2 atmospheres and dragged down to the aorta. The rats were killed 3 weeks after the injury. The carotid intima/media ratio was calculated. Intimal hyperplasia after carotid balloon-induced injury in the fatty Zucker rats was significantly reduced in the group treated with rosiglitazone (0.18 +/- 0.29) compared with the untreated group (0.97 +/- 0.13; P<.01). Plasma glucose, triglyceride, and insulin levels were elevated, indicative of the presence of insulin resistance; rosiglitazone treatment significantly reduced insulin and triglyceride levels without decreasing glucose. Rosiglitazone treatment also reduced, but to a lesser extent, the intimal hyperplasia in the lean Zucker rats (0.57 +/- 0.10 vs 1.06 +/ 0.12 treated and untreated, respectively; P<.01); however, it had no effect on insulin, triglyceride, or glucose levels in this group. The intimal hyperplasia in the fatty Zucker rats treated with fenofibrate was not reduced compared with controls (0.84 +/- 0.26 vs 0.97 +/- 0.13, respectively); fenofibrate reduced insulin and triglyceride, but not glucose levels, in these animals. CONCLUSIONS: The PPAR-gamma ligand rosiglitazone, but not the PPAR-alpha ligand fenofibrate, decreases intimal hyperplasia following balloon injury in both fatty and lean Zucker rats. This effect of the PPAR-gamma ligand was independent of glycemia, insulin, and lipid levels, and was more pronounced in insulin-resistant rats. PMID- 14740080 TI - Right atrial and ventricular thrombus infiltrated with osteoblastic osteosarcoma. AB - The presenting signs of osteogenic osteosarcoma are commonly pain, local swelling, local warmth, pathologic fracture, and metastatic disease. Deep venous metastasis of osteoblastic osteosarcoma is most often a postmortem diagnosis. This paper describes the case of a previously healthy 18-year-old woman who presented with dyspnea and lower extremity edema. This is a rare, and to our knowledge, a previously unreported case of right atrial and ventricular tumor thrombus infiltrated with osteoblastic osteosarcoma. PMID- 14740081 TI - Reversal of refractory congestive heart failure after thiamine supplementation: report of a case and review of literature. AB - Patients with refractory heart failure comprise a very important subgroup of patients with congestive heart failure. Before assuming that this condition simply reflects advanced, perhaps terminal, myocardial dysfunction, potentially reversible factors should be sought carefully. We describe a 58-year-old Hispanic man with a diagnosis of idiopathic dilated cardiomyopathy who presented with symptoms of severe congestive heart failure, glossitis, and peripheral neuropathy. His hemodynamic profile was characterized by refractory low-output cardiac failure and decreased vascular resistance. Thiamine deficiency was documented by a high thiamine pyrophosphate effect. His clinical condition was quickly reversed with thiamine administration. This response to thiamine administration supports the diagnosis and indicates that thiamine deficiency may play an important etiologic role in the deterioration of cardiac function in some patients with congestive heart failure. PMID- 14740082 TI - Advanced backcross QTL analysis of a Lycopersicon esculentum x L. pennellii cross and identification of possible orthologs in the Solanaceae. AB - In this study, the advanced backcross QTL (AB-QTL) mapping strategy was used to identify loci for yield, processing and fruit quality traits in a population derived from the interspecific cross Lycopersicon esculentum E6203 x Lycopersicon pennellii accession LA1657. A total of 175 BC(2) plants were genotyped with 150 molecular markers and BC(2)F(1) plots were grown and phenotyped for 25 traits in three locations in Israel and California, U.S.A. A total of 84 different QTLs were identified, 45% of which have been possibly identified in other wild-species derived populations of tomato. Moreover, three fruit-weight/size and shape QTLs ( fsz2b.1, fw3.1/ fsz3.1 and fs8.1) appear to have putative orthologs in the related solanaceous species, pepper and eggplant. For the 23 traits for which allelic effects could be deemed as favorable or unfavorable, 26% of the identified loci had L. pennellii alleles that enhanced the performance of the elite parent. Alleles that could be targeted for further introgression into cultivated tomato were also identified. PMID- 14740085 TI - Identification and molecular characterization of a novel y-type Glu-Dt 1 glutenin gene of Aegilops tauschii. AB - A novel y-type high-molecular-weight glutenin subunit possessing a slightly faster mobility than that of subunit 1Dy12 in SDS-PAGE, designated 1Dy12.1(t) in Aegilops tauschi, was identified by one- and two-dimensional gel and capillary electrophoresis. Its coding gene at the Glu-D(t) 1 locus was amplified with allele-specific-PCR primers, and the amplified products were cloned and sequenced. The complete nucleotide sequence of 2,807 bp containing an open reading frame of 1,950 bp and 857 bp of upstream sequence was obtained. A perfectly conserved enhancer sequence and the -300 element were present at positions of 209-246 bp and 424-447 bp upstream of the ATG start codon, respectively. The deduced mature protein of 1 Dy12.1(t) subunit comprised 648 amino acid residues and had a Mr of 67,518 Da, which is slightly smaller than the 1Dy12 (68,695 Da) but larger than the 1Dy10 (67,495 Da) subunits of bread wheat, respectively, and corresponds well with their relative mobilities when separated by acid-PAGE. The deduced amino acid sequence indicated that the 1Dy12.1(t) subunit displayed a greater similarity to the 1Dy10 subunit, with only seven amino acid substitutions, suggesting that this novel gene could have positive effect on bread-making quality. A phenetic tree produced by nucleotide sequences showed that the x- and y-type subunit genes were respectively clustered together and that the Glu-D(t) 1y12.1 gene of Ae. tauschii is closely related to other y type subunit genes from the B and D genomes of hexaploid bread wheat. PMID- 14740087 TI - QTL mapping for resistance against non-parasitic leaf spots in a spring barley doubled haploid population. AB - Phenotypic variability for resistance against non-parasitic leaf spots (NPLS) has been observed between varieties. For the genetic characterization of NPLS resistance, a population with 430 doubled haploid (DH) lines was developed from the cross between the NPLS-resistant Hordeum vulgare breeding line IPZ24727 and the NPLS-sensitive barley cultivar Barke. A molecular map was constructed based on 164 AFLPs, 30 SSRs and one STS marker derived from the mlo gene. Field trials were performed over four environments in which NPLS and other agronomic traits were assessed. Estimates of genotypic variance were highly significant for NPLS. Moreover, no transgression was found for the trait. Quantitative trait loci (QTLs) for NPLS resistance were mapped in the DH population on chromosomes 1H, 4H, and 7H, with the most important effect on chromosome 4H. The QTLs for NPLS explained together 39% of the phenotypic and 49% of the genotypic variance, thereby showing additive gene action. Consequently, marker-assisted selection for improving NPLS resistance is possible. PMID- 14740086 TI - Development of PCR-based SNP markers for rice blast resistance genes at the Piz locus. AB - We assessed the utility of single-nucleotide polymorphisms (SNPs) and small insertion/deletion polymorphisms (InDels) as DNA markers in genetic analysis and breeding of rice. Toward this end, we surveyed SNPs and InDels in the chromosomal region containing the Piz and Piz-t rice blast resistance genes and developed PCR based markers for typing the SNPs. Analysis of sequences from a blast-susceptible Japanese cultivar and two cultivars each containing one of these genes revealed that SNPs are abundant in the Piz and Piz-t regions (on average, one SNP every 248 bp), but the number of InDels was much lower. The dense distribution of SNPs facilitated the generation of SNP markers in the vicinity of the genes. For typing these SNPs, we used a modified allele-specific PCR method. Of the 49 candidate allele-specific markers, 33 unambiguously and reproducibly discriminated between the two alleles. We used the markers for mapping the Piz and Piz-t genes and evaluating the size of DNA segments introgressed from the Piz donor cultivar in Japanese near-isogenic lines containing Piz. Our findings suggest that, because of its ability to generate numerous markers within a target region and its simplicity in assaying genotypes, SNP genotyping with allele specific PCR is a valuable tool for gene mapping, map-based cloning, and marker assisted selection in crops, especially rice. PMID- 14740088 TI - Identification of SNPs in the waxy gene among glutinous rice cultivars and their evolutionary significance during the domestication process of rice. AB - Common non-waxy ( Wx) rice cultivars contain two different alleles at the waxy locus, designated Wx(a) and Wx(b), which encode different levels of granule-bound starch synthases and are hence involved in the control of endosperm amylose content. The Wx(a) allele was predominant in non-waxy indica cultivars, whereas the Wx(b) allele was common to the non-waxy japonica variety. Recently, some of the molecular mechanisms underlying the differentiation of Wx(a) from Wx(b) have been characterized. One structural difference between these two alleles was shown to be due to alternative splicing caused by a single-base substitution (AG GT to AG TT) at a donor site of the first intron within the Wx gene. In the case of waxy ( wx) rice, it was not possible to distinguish whether the each wx allele was derived from Wx(a) or Wx(b) alleles by phenotypic analysis. However, we succeeded in developing a derived cleaved amplified polymorphic sequence (dCAPS) marker for the detection of the one-base splicing mutation without the need for sequencing. A mismatch primer was used to generate a restriction site in the Wx(a) allele (AGGT) but not in the Wx(b) allele (AGTT). Three hundred fifty-three waxy rice strains that are widely found in Asia were then employed for analysis using this dCAPS marker. Our findings suggested that waxy rice strains have both Wx(a)- and Wx(b)-derived alleles, but that the Wx(b)-derived allele was predominant, and its distribution was independent of indica- japonica differentiation. The wild relatives of cultivated rice all possessed the AGGT allele. It was concluded that the waxy mutations, and the corresponding rice cultivation, originated from japonica during the evolution and domestication process of rice and was preferentially selected by most Asian peoples. PMID- 14740089 TI - Optimal sampling of a population to determine QTL location, variance, and allelic number. AB - In a population intended for breeding and selection, questions of interest relative to a specific segregating QTL are the variance it generates in the population, and the number and effects of its alleles. One approach to address these questions is to extract several inbreds from the population and use them to generate multiple mapping families. Given random sampling of parents, sampling strategy may be an important factor determining the power of the analysis and its accuracy in estimating QTL variance and allelic number. We describe appropriate multiple-family QTL mapping methodology and apply it to simulated data sets to determine optimal sampling strategies in terms of family number versus family size. Genomes were simulated with seven chromosomes, on which 107 markers and six QTL were distributed. The total heritability was 0.60. Two to ten alleles were segregating at each QTL. Sampling strategies ranged from sampling two inbreds and generating a single family of 600 progeny to sampling 40 inbreds and generating 40 families of 15 progeny each. Strategies involving only one to five families were subject to variation due to the sampling of inbred parents. For QTL where more than two alleles were segregating, these strategies did not sample QTL alleles representative of the original population. Conversely, strategies involving 30 or more parents were subject to variation due to sampling of QTL genotypes within the small families obtained. Given these constraints, greatest QTL detection power was obtained for strategies involving five to ten mapping families. The most accurate estimation of the variance generated by the QTL, however, was obtained with strategies involving 20 or more families. Finally, strategies with an intermediate number of families best estimated the number of QTL alleles. We conclude that no overall optimal sampling strategy exists but that the strategy adopted must depend on the objective. PMID- 14740090 TI - Quantitative trait loci affecting plant regeneration from protoplasts of Brassica oleracea. AB - Quantitative trait loci (QTLs) controlling the plant-regeneration ability of Brassica oleracea protoplasts were mapped in a population of 128 F(2) plants derived from a cross between the high-responding, rapid-cycling line and a low responding, broccoli breeding line of B. oleracea. A modified bulked segregant analysis with AFLP markers identified two QTLs for plant regeneration. In a multiple regression analysis, the two QTLs explained 83% of the total genetic variation for regeneration recorded 15 weeks after initial transfer of microcalli to regeneration medium. Both QTLs showed additive effects, and the alleles contributing to the high regeneration frequencies were derived from the high responding, rapid-cycling line. Using microsatellites with known location, the two QTLs were mapped to linkage groups O2 and O9 on the map published by Sebastian et al. [(2000) Theor Appl Genet 100:75-81] or to chromosomes C8 and C7 on the map published by Saal et al. [(2001) Theor Appl Genet 102:695-699]. QTLs for the early flowering trait of the rapid-cycling parent have previously been mapped to the same two linkage groups. Association between flowering time and regeneration ability was, however, not found in the present material, indicating that plant-regeneration ability can be transferred between cultivars independently of the early flowering trait. The detection of two major QTLs for plant regeneration in B. oleracea may provide the initial step towards the identification of markers suitable for marker-assisted selection of regeneration ability. PMID- 14740091 TI - Genetic diversity in cultivated plants-loss or stability? AB - Human activities like urbanisation, the replacement of traditional agriculture systems by modern industrial methods or the introduction of modern high-yielding varieties may pose a danger to the biological diversity. Using microsatellite markers, we analysed samples of cultivated wheat ( Triticum aestivum L.) collected over an interval of 40-50 years in four comparable geographical regions of Europe and Asia. No significant differences in both the total number of alleles per locus and in the PIC values were detected when the material collected in the repeated collection missions in all four regions were compared. About two thirds of the alleles were common to both collection periods, while one-third represented collection mission-specific alleles. These findings demonstrate that an allele flow took place during the adaptation of traditional agriculture to modern systems, whereas the level of genetic diversity was not significantly influenced. PMID- 14740092 TI - [Clinical MR at 3 Tesla: current status]. AB - Clinical MRI is mostly performed at field strengths up to 1.5 Tesla (T). Recently, approved clinical whole-body MR-systems with a field strength of 3 T became available. Its installation base is more rapidly growing than anticipated. While site requirements and operation of these systems do not differ substantially from systems with lower field strength, there are differences in practical applications. Imaging applications can use the gain in signal-to-noise for increased spatial resolution or gain in speed. This comes at a trade off in increased sensitivity to field inhomogeneities and changes in relaxation times, which lead to changes in image contrast. The benefit of high field for spectroscopy consists in increased signal-to-noise-ratio and improvement in frequency resolution. The increase in energy deposition necessitates the use of special strategies to reduce the specific absorption rate (SAR). This paper summarizes the current state of MR at 3 T. PMID- 14740093 TI - [Clinical high- and ultrahigh-field MR and its interaction with biological systems]. AB - The field strength of the static field in MRI has increased from 0.015 to 12 Tesla (T) during the last 25 years, which is about an 800 fold increase. In addition to low- and high field systems (1.5-4 T), ultra-high field systems with field strengths above 4 T are now available for human MRI. The extension of non significant risk status for clinical fields up to 8 T by the FDA in July 2003 facilitates the further growth of this technology. The increase in field strength creates the need for a better understanding of the safety challenges to ensure safety for human imaging applications. This encompasses understanding the effects of the strong magnetic field at the atomic and molecular level and from biological tissue to organ systems. Moreover, in addition to the effects of a static magnetic field, the effects of radio-frequency- and gradient-fields have to be considered. This paper reviews the safety relevant issues for high- and ultrahigh field MR. PMID- 14740094 TI - [Feasibilities and limitations of high field parallel MRI]. AB - In medical magnetic resonance imaging (MRI) it is standard to use MR scanners with a field strength of 1.5 Tesla. Recently, an ongoing development to higher field strength can be observed and a new clinical standard at 3.0 Tesla seems to be established. High field MRI with its intrinsic higher signal to noise ratio (SNR) can enable new applications of MRI in medical diagnosis, or can serve to improve existing methods. It is important to note, that the use of high field MRI is not without its limitations. Besides the SNR, other unwanted effects increase with a higher field strength. Without correction, these high field problems cause a serious loss in image quality. An elegant way to address these problems is the use of parallel imaging. In many clinical applications, parallel MRI (pMRI) is part of the standard protocol, because pMRI can enhance virtually every MRI application, without necessarily affecting the contrast behavior of the underlying imaging sequence. In high field MRI, besides the speed advantage of pMRI, the positive influence on high field specific problems and therefore on the image quality will be of major importance. PMID- 14740095 TI - [Use of contrast agent in high-field MRI (3 T)]. AB - The basic diagnostic efficacy of MR contrast medium in the evaluation of primary brain tumors and its clinical usefulness in the detection of brain metastases with single and cumulative triple-dose was compared using a high-field 3 T MR unit and a 1.5 T MR unit. Additionally, the effect of contrast agent on high resolution MR venography based on the BOLD effect was evaluated at both field strengths. Tumor-brain contrast after gadodiamide administration, as assessed by means of statistical evaluation of MP-RAGE scans and T1-SE images, was significantly higher at 3 T than at 1.5 T. The subjective assessment of cumulative triple-dose 3 T images obtained the best results in the detection of brain metastases, followed by 1.5 T cumulative triple-dose enhanced images. Due to higher spatial resolution, contrast-enhanced MR venography at 3 T showed more details in and around tumors than at 1.5 T, additionally enhanced by stronger susceptibility weighting and higher signal-to-noise ratio at 3 T. In summary, administration of gadolinium-based contrast agent produces higher contrast between tumor and normal brain at 3 T than at 1.5 T, helps to detect more cerebral metastases at 3 T than at 1.5 T in single and cumulative triple dose, and improves MR venography at 3 T with increase in spatial resolution within the same measurement time, thus providing more detailed information. PMID- 14740096 TI - [Hospital radiology--what is the way forward in view of the structural changes in health care]. PMID- 14740097 TI - [Unclear ascites in a young male patient]. PMID- 14740099 TI - New Early Cretaceous fossil from China documents a novel trophic specialization for Mesozoic birds. AB - We report on a new Mesozoic bird, Longirostravis hani, from the Early Cretaceous Jehol Biota of northeastern China. The new taxon has a long, slender rostrum and mandible, and a small number of rostralmost teeth. Postcranial characters such as a furcular ramus wider ventrally than dorsally, a centrally concave proximal margin of the humeral head, and a minor metacarpal that projects distally more than the major metacarpal, support the placement of Longirostravis within euenantiornithine Enantiornithes, the most diverse clade of Mesozoic birds. The morphology of the skull, however, suggests that Longirostravis had a probing feeding behavior, a specialization previously unknown for Enantiornithes. Indeed, this discovery provides the first evidence in support of the existence of such a foraging behavior among basal lineages of Mesozoic birds. PMID- 14740098 TI - Early human development: new data raise important embryological and ethical questions relevant for stem cell research. AB - Recent research has considerably changed our views about the developmental biology of early mammalian embryos compared with the ideas that were predominant throughout the previous 30 years or so. New data obtained recently suggest that the mammalian embryo uses traits of axes determination mechanisms that are not too different from the modes used by other vertebrates. In particular, it appears that asymmetry cues derived from the oocyte cytoplasm and modified/specified during sperm penetration appear to be crucial in normal embryogenesis, rather than the environmental influences exerted, e.g. at embryo implantation in the uterus. On the other hand, recent advances in research on the equivalents of a Spemann-Mangold organizer in species other than amphibia (including mammals) provide a background for new discussions of early embryonic patterning (axis formation) processes in the embryonic disc. In combination, these new views appear to be of considerable interest in the debate on the developmental properties and the ethical status of embryonic stem cells. The present review focuses specifically on the new aspects of axis determination and pattern formation processes in early mammalian embryos and relates this to questions about the developmental potential of embryonic stem cells (totipotency vs pluripotency/omnipotency), i.e. facts that appear to be worth considering in the recent debate about the ontological status of the early human embryo as well as of human embryonic stem cells. PMID- 14740100 TI - The eyes of oilbirds (Steatornis caripensis): pushing at the limits of sensitivity. AB - An extreme example of a low light-level lifestyle among flying birds is provided by the oilbird, Steatornis caripensis (Steatornithidae, Caprimulgiformes). Oilbirds breed and roost in caves, often at sufficient depth that no daylight can penetrate, and forage for fruits at night. Using standard microscopy techniques we investigated the retinal structure of oilbird eyes and used an ophthalmoscopic reflex technique to determine the parameters of these birds' visual fields. The retina is dominated by small rod receptors (diameter 1.3+/-0.2 microm; length 18.6+/-0.6 microm) arranged in a banked structure that is unique among terrestrial vertebrates. This arrangement achieves a photoreceptor density that is the highest so far recorded (approximately 1,000,000 rods mm(-2)) in any vertebrate eye. Cone photoreceptors are, however, present in low numbers. The eye is relatively small (axial length 16.1+/-0.2 mm) with a maximum pupil diameter of 9.0+/-0.0 mm, achieving a light-gathering capacity that is the highest recorded in a bird (f-number approximately 1.07). The binocular field has a maximum width of 38 degrees and extends vertically through 100 degrees with the bill projecting towards the lower periphery; a topography that suggests that vision is not used to control bill position. We propose that oilbird eyes are at one end of the continuum that juxtaposes the conflicting fundamental visual capacities of sensitivity and resolution. Thus, while oilbird visual sensitivity may be close to a maximum, visual resolution must be low. This explains why these birds employ other sensory cues, including olfaction and echolocation, in the control of their behaviour in low-light-level environments. PMID- 14740101 TI - Influence of CO2 on the micro-structural properties of spider dragline silk: X ray microdiffraction results. AB - The mechanical properties of spider dragline fibres are altered by CO(2) exposure under anaesthetizing conditions during the spinning process. In order to relate these macroscopic changes to a microscopic model, the extrusion of dragline silk was studied by synchrotron radiation microdiffraction. A brief exposure of a female Nephila senigalensis spider to CO(2 )results after an incubation time of less than 7 min in the extrusion of a thread (two fibres) swollen with water. The data are interpreted for a model of crystalline beta-sheet domains containing nanofibrils, which reinforce a network of protein chains. The protein network absorbs water, leaving the nanofibrils unaffected. A continuous flow of CO(2) results in a co-extrusion of a dragline thread and an isotropic silk fraction, which probably has a glycine-rich sequence. Long CO(2) exposure reduces the axial alignment of nanofibrils, presumably due to a partial destruction of the amorphous network. PMID- 14740102 TI - A new C12 alcohol identified as a sex pheromone and a trail-following pheromone in termites: the diene (Z,Z)-dodeca-3,6-dien-1-ol. AB - The diunsaturated C12 alcohol (Z,Z)-dodeca-3,6-dien-1-ol (dodecadienol) has been characterized by GC-MS and FTIR as a novel releaser pheromone in termites. This alcohol identified in Ancistrotermes pakistanicus (Termitidae, Macrotermitinae) possesses a double pheromonal function which again illustrates the chemical parsimony of termites compared with other social insects. In workers, dodecadienol elicits trail-following at a very low concentration (activity threshold at 0.1 pg/cm of trail); in male alates it induces trail-following at a low concentration (1-10 pg/cm) and sexual attraction at a higher concentration (about 1 ng). Traces of the monounsaturated C12 alcohol (Z)-dodec-3-en-1-ol (dodecenol), known as a trail pheromone of several Macrotermitinae, were also found in the sternal gland extracts of A. pakistanicus, although only dodecadienol was present at the surface of the sternal gland. Workers of A. pakistanicus are not sensitive to dodecenol, but they are as sensitive to dodecatrienol as to dodecadienol. However, in the study area (Vietnam), A. pakistanicus is living in sympatry only with those Macrotermitinae using dodecenol as a trail pheromone, the foraging populations therefore being well isolated through their respective trail pheromones. The presence of three types of unsaturated C12 alcohols as releaser pheromones in the only Macrotermitinae subfamily is discussed, and a possible biosynthetic pathway from linoleic acid is proposed for dodecadienol. PMID- 14740103 TI - A unique strategy of host colony exploitation in a parasitic ant: workers of Polyrhachis lama rear their brood in neighbouring host nests. AB - The formicine ant Polyrhachis lama is a social parasite, exploiting its ponerine host ant species Diacamma sp. In most social parasitic associations, the parasitic species are closely related to their host species group, evolving directly from independent ancestors of the host species. However, in the Polyrhachis lama- Diacamma sp. association, the associated species belong to different ant subfamilies. Based on preliminary field surveys, we had presumed that P. lama might have given up its reproductive division of labour, i.e. workers would be able to produce males as well as workers and females parthenogenetically. In this study, this hypothesis was disproved: Polyrhachis lama workers cannot be fertilized and are only able to produce males. In the host parasite association originating from nests possessing a P. lama queen, workers penetrate surrounding Diacamma sp. nests, carrying brood for rearing within these satellite nests. In this peculiar way, a single P. lama colony is able to exploit several Diacamma sp. colonies simultaneously. PMID- 14740104 TI - Multiple paternity in Rana dalmatina, a monogamous territorial breeding anuran. AB - Polyandry and sperm competition in anurans have rarely been documented. We investigated the genetic paternity inferred from allozyme variations in 650 tadpoles from four natural ponds in a territorial breeding anuran, Rana dalmatina. Multiple paternity was demonstrated, although R. dalmatina is regarded as a monogamous species. Polyandrous mating was not a common event, occurring only in 17.9% of clutches, with no significant differences among clutches. The proportions of tadpoles fathered by a second male did not significantly differ among ponds, showing that multipaternity was not restricted to a single site. Such a polyandry may result from synchronous multiple amplexus and should reduce the heterozygote deficit related to the breeding-pond fidelity usually exhibited by most anurans. PMID- 14740107 TI - Sudden cardiac death is a very serious matter. PMID- 14740108 TI - [Gastroesophageal reflux (GER)--cause of otitis media with effusion in childhood?]. PMID- 14740109 TI - [Paraganglioma in the area of the head and neck. A review of molecular genetic research]. AB - Paragangliomas of the head and neck region are usually benign tumors that develop from chemoreceptors of paraganglionic origin in the majority of patients. These receptors play an important role in sensing and regulation of the blood CO(2) level. Genetic alterations in the mitochondrial enzyme complex II (SDH), which is involved in respiratory chain and citric acid cycle reactions, have been shown to lead to sporadic as well as familial cases of these tumors. The gene encoding the subunit SDHD shows mutations in up to 50% of these cases. In addition, loss of heterozygosity (LOH) was demonstrated in these tumor samples and has been shown to be connected with oncogenesis of paragangliomas. Thus, SDHD is the first known tumor suppressor gene encoding a mitochondrial protein. In this article we summarize the current state of knowledge concerning the development of paragangliomas. PMID- 14740110 TI - [Effect of high frequency application on the tongue base. In vitro results of "radiofrequency application" on the porcine tongue base]. AB - BACKGROUND AND OBJECTIVE: A newly developed radio-frequency monopolar needle electrode was evaluated in vitro on porcine tongues. PATIENTS AND METHODS: Porcine tongues with different tissue temperatures (20+/-1) degrees C and (32+/ 2) degrees C were coagulated for 90 s. In a second step, 23 coagulations at 7 W were applied (34+/-2) degrees C. RESULTS: The volume of the lesion correlated well with increasing temperature. In step two, the mean energy was 238 J and the mean volume of the lesion was 507 mm(3) (100+/-15)%. Prolonged energy application did not correlate with a larger volume of the lesion. CONCLUSION: The new radio frequency needle electrode can reduce tongue volume in a precise and controlled manner (SD+/-15%). PMID- 14740111 TI - [Histomorphologic and salivary gland scintigraphic findings in radiation-induced sialadenitis due to fractionated irradiation of the head and neck region of rats. A model for evaluating potentially radioprotective substances]. AB - INTRODUCTION: The aim of this study was to correlate structural, histomorphological damage of the salivary gland with scintigraphic findings during fractioned radiotherapy. METHODS: The head and neck area of 27 WAG/RijH rats was irradiated with (60)Co-gamma-rays (60 Gy/30f/6 weeks). A port-system was implanted and (99m)Tc-pertechnetat applied at different stages of irradiation (0, 16, 30, 46, 60 Gy and 6 months post irradiation). RESULTS: After the application of 16 Gy an intra- and extra-cellular oedema developed in the salivary glands. The progressive vacuolisation (30 Gy) passed over into lipomatosis (46 Gy) and necrosis (60 Gy) in the parotid and mandibular glands. Six months after irradiation treatment, the chronic histomorphological damage corresponded to stage II according to Seifert. The corresponding loss in gland function was 13% (16 Gy); 26% (30 Gy); 57% (46 Gy); 75% (60 Gy) and 66.5% (6 months post irradiation). CONCLUSION: This animal model demonstrates the correlation between histomorphological and scintigraphic findings. PMID- 14740112 TI - [Surgical therapy of malignant melanoma of the external ear]. AB - METHOD: The data from our patients of the last 10 years were evaluated in relation to survival and recurrence. RESULTS: Very different surgical therapy-in some cases due to refusal of the therapy suggested-was performed in eight patients (including wedge shaped excision as well as ablation of the auricle). The mean survival ranged from 40 months up to the present. One patient died due to a local recurrence and regional and systemic metastases. One patient experienced local recurrence but is still well 16 months after he underwent surgical revision. Two patients died independently of the tumor. DISCUSSION: Comparison of our results with those of other studies confirms that the extent of the applied resection has of minor effect on recurrence and survival for this disease. Tumor thickness is the most important independent prognostic factor. PMID- 14740114 TI - ["Off-label use" of botulinum toxin preparations in treatment of spasmodic dysphonia. Position of the German Society of Phoniatrics and Pediatric Audiology]. PMID- 14740113 TI - [Diagnosis with (18)F-FDG PET scan after larynx preservation by primary radiochemotherapy]. AB - PATIENTS AND METHODS: In a group of 20 patients undergoing chemoradiation for larynx organ preservation after diagnosis of laryngeal and hypopharyngeal carcinoma, (18)F-fluordeoxyglucose positron emission tomography ((18)F-FDG-PET) was performed before the start of therapy. After i.v. application of 240 MBq FDG, a dynamic PET in 3-D-mode was performed over 90 min (Siemens CTI ECAT EXACT HR(+)). Analysis was done visually and semiquantitatively (60-90 min p.i.) following iterative reconstruction. Additional (18)F-FDG-PET investigations were done and correlated with the clinical outcome in 16/20 patients at 3 months and in 14/20 patients at 6 months after the end of therapy. RESULTS: In 17/20 patients (85%), the preclinical (18)F-FDG-PET correlated well with the histologically confirmed primary tumor. Three cases were false negatives. In one case this was due to an increased glucose value (203 mg%). After 3 months, 8/13 (62%) patients showed a positive correlation between clinical and PET results (sensitivity 100%, specificity 70%). After 6 months, 9/11 (82%) patients presented clinically normal PET results. PET results were false negative in one case (sensitivity 67%, specificity 88%). CONCLUSION: The data of our trial slightly reduce the enthusiasm of early (18)F-FDG-PET detection of residual disease after chemoradiation in resectable laryngeal or hypopharyngeal cancer. Further trials should optimize the calculation integrating the exact quantification of glucose metabolism with the aim of improving sensitivity and specificity. PMID- 14740115 TI - [Post-partum rapidly growing maxillary sinus tumor]. PMID- 14740116 TI - [First use of botulinum toxin type B in ENT patients with secondary therapy failure of botulinum toxin type A]. AB - The first botulinum toxin type B (BT-B) formulation, NeuroBloc, has been licensed in Germany for the treatment of cervical dystonia since March 2001. This allows the treatment of patients with secondary failure of botulinum type A (BT-A) injections for the first time. Three patients with such a secondary failure were successfully treated with BT-B. A total of 1,000 mouse units (MU) NeuroBloc per eye were injected in a patient with blepharospasm. The treatment was effective for 4 months. In a patient with spasmodic dysphonia, 250 MU were injected into each vocal cord. This was effective for 3.5 months. The third patient suffered from bilateral Frey's syndrome. A total of 7,500 MU were used on the right side and 6,000 MU on the left side to stop the gustatory sweating for 9 months. Overall, BT-B showed similar intervals of effectiveness as BT-A. Side effects were not seen using the above dosage regimes. PMID- 14740118 TI - [Hyperbaric oxygen therapy after unilateral idiopathic sudden deafness]. AB - The use of hyperbaric oxygenation therapy (HBOT) after acute one-sided deafness is a treatment option if conventional methods fail. Five cases have been reported in which an improvement in hearing after HBOT was achieved following unsuccessful conventional therapy. In view of this, and after a careful study of the literature, we suggest that timely treatment with HBOT should be used in every case of unilateral idiopathic deafness. PMID- 14740117 TI - [Reconstruction of the frontal sinus with a calvarium split galea periosteum transplant after inflammatory complications]. AB - After reconstruction of the frontal sinus with alloplastic material, with or without obliteration of the lumen, inflammatory complications may occur. The authors present a plastic-surgical functional reconstruction technique for revision surgery in cases of inflammatory complications following frontal sinus implantation of alloplastic material, by means of a modified autologous calvarian split graft. The inflammatory complications after primary reconstruction or obliteration of the frontal sinus occurred after a period of 1-10 years. The previously implanted material was removed from the frontal sinus and the anterior wall was reconstructed with a combined calvarian split/galea periost-transplant. During follow-up, which ranged from 1-5 years, no further inflammatory complications occurred. Furthermore, an appealing cosmetic result was achieved in all cases. We conclude that the functional and aesthetic result depends primarily on (1) embedding and protection of the graft by an additional covering with a galea-periost-flap, and (2) reconstruction of the frontal drainage pathways. PMID- 14740119 TI - [Late auditory potentials (NC-ERP) in children with symptoms of auditory processing and perception disorder. With and without attention deficit disorder]. AB - BACKGROUND: Auditory processing disorders (APD) result from dysfunctions of processes dedicated to audition and affect processing of information in the auditory modality. Children with APD exhibit symptoms similar to those with attention deficit disorders (ADHD). With regard to therapeutic strategies it seems mandatory to employ diagnostic procedures able to differentiate between APD and ADHD. Recently it was found that auditory evoked neutral condition event related potentials (NC-ERP) correlate with results from psychoacoustic measures suitable to unveil dysfunctions of auditory processing, thus confirming APD. Here we investigated whether NC-ERP may be used as a tool to differentiate between APD and ADHD. METHODS: In a retrospective analysis, NC-ERP data from 99 children with (+ADHD) and 43 without ADHD symptoms (-ADHD) were examined. All patients suffered from symptoms consistent with information processing deficits in the auditory modality. Analysis of variance was applied. RESULTS: +ADHD children and -ADHD children did not show a significantly different distribution pattern of pathological NC-ERP. DISCUSSION: Since the probability of pathological NC-ERP is equally distributed in children with or without ADHD, all of whom suffered from auditory processing deficits, it seems fair to conclude that NC-ERP are not suitable to differentiate between APD and ADHD. This may be due to methodological limitations. Alternatively, APD and ADHD may indeed be overlapping entities with a common yet unidentified origin. PMID- 14740120 TI - [Facial nerve rehabilitation after cerebellopontile angle surgery]. PMID- 14740121 TI - [Imaging modalities in the preoperative staging of rectal carcinoma]. AB - Thorough clinical examination, endorectal ultrasound, and magnetic resonance tomography (MRI) are decisive tools in the pretherapeutic work-up of patients with rectal cancer. Depth of infiltration to the rectal wall as well as involvement of perirectal lymph nodes by the tumor are the key questions to be answered. To receive adequate information from imaging procedures, the right questions need to be asked. The extent of invasion of the rectal wall and exact location of tumor infiltration to neighboring structures can be demonstrated by MRI very well, particularly if imaging planes are acquired at 90 degrees to the position of the rectum. Recent developments in computed tomography (CT) using isotrope voxels allow three-dimensional reconstructions of tumors without loss of imaging quality. Assessment of the primary tumor and its nodal metastases after preoperative radio-(chemo-)therapy is still seriously limited. Fusion of positron emission tomography and CT could be a step towards solving the problem of response assessment in the near future. PMID- 14740122 TI - [Reconstructive surgery after anterior resection of the rectum]. AB - Sphincter preservation is a goal in the treatment of rectal cancer and should be considered in all patients with intact analsphincter. For tumors of the upper rectum, this is easily achieved by end-to-end decendorectostomy. Total mesorectal excision is obligatory for tumors of the mid and lower third of the rectum. Reconstruction of intestinal continuity can be achieved by colonal anastomosis, if the oncologic situation allows a sphincter-preserving procedure. Creation of a colonic J-pouch can improve functional results. This leads to a reduction of stool frequency and urgency without negative effects on continence and complication rate. The functional effects are maximal during the initial months, but are still significant up to 9 years postoperatively. The pouch should not exceed a length of 6 cm in order to prevent evacuation disorders. The transverse coloplasty pouch may allow pouch reconstruction in patients in whom this is currently impossible, but long-term follow-up is not yet available. PMID- 14740123 TI - [Anal sphincter replacement]. AB - Presently, deep rectal carcinoma is usually treated by deep anterior rectal resection and colonal anastomosis. Abdominoperineal resection is needed only for the very few patients whose tumors infiltrate the pelvic base or sphincter musculature. This means the loss of normal anal function and thus of normal defecation. Many patients find the idea of a stoma unacceptable. In our experience, the construction of a functional neoanus after abdominoperineal rectal resection is a suitable option for patients in good general health and who are highly motivated. PMID- 14740124 TI - [Quality of life after rectal surgery]. AB - During recent years, quality of life has become an increasingly important parameter in the outcome assessment of therapeutic protocols in oncology. In rectal cancer treatment, the influence of different surgical techniques on quality of life can be evaluated and compared by a variety of standardized and clinically validated instruments. As yet, sphincter-saving surgical techniques were considered to result in higher quality of life than those sacrificing the sphincter. Recent data from clinical trials indicates that stoma patients have comparable or even higher quality of life than patients of sphincter-saving surgery. However, prospective clinical trials assessing not only classic quality of life items but also unconventional parameters, i.e., patient expectations etc., are still lacking. Thus, detailed interpretation of the differences assessed by the classic quality of life scores is profoundly hampered. The realisation of such a prospective clinical trial will be a further important step to improve quality of life in patients suffering from rectal cancer by improving not only surgical techniques but also appropriate psychological patient management. PMID- 14740125 TI - [Principles of postoperative therapy in rectal carcinoma]. AB - In stage II (T3-4N0) and III (TxN1-2) rectal cancer, adjuvant radiochemotherapy improves overall survival and decreases the rate of local failure compared to only surgical therapy and is regarded as standard for patients with carcinoma of the lower and intermediate rectum. (Preoperative) radiotherapy also decreases the local failure rate following total mesorectal excision. Postoperative radiotherapy has no proven influence on distant metastasis rates or on overall survival. In adjuvant therapy, continuous infusion of 5-flourouracil compared to bolus application increases long-term survival. However, additional administration of leucovorin or levamisole results in increased toxicity and not improved survival. Results of randomized trials of adjuvant therapy with new drugs such as capecitabine, UFT, irinotecan, or oxaliplatin are not yet available. These drugs should not be used outside clinical trials. Elderly patients benefit from adjuvant therapy to the same extent as younger patients and should receive adjuvant radiochemotherapy, if no contraindication exists. PMID- 14740126 TI - [Locally recurrent rectal carcinoma]. AB - There is no doubt that surgical treatment with curative intent is warranted in patients with recurrent colorectal cancer. This is also true for locally recurrent rectal cancer. Operative procedures are demanding but may be curative if complete excision can be achieved. Preoperative radiochemotherapy has helped to improve resectability, but operative morbidity is still high. Procedure related mortality on the other hand is low, at around 3%, and 25% 5-year survival can be expected after complete excision. PMID- 14740127 TI - [Graduated compression stockings in surgery -- optional or obligatory?]. AB - BACKGROUND: Graduated compression stockings (GCS) can effectively reduce postoperative deep vein thrombosis (DVT) and their use is recommended by expert committees. However, it appears that GCS are not frequently used. The objectives of this study are to evaluate the customary use of GCS in surgical settings in the City of Hamburg, Germany, and to present evidence on the effectiveness of GCS. METHODS: A questionnaire on the use of thromboprophylaxis was sent to 48 surgeons in Hamburg. In addition, a comprehensive search for randomized controlled trials, reviews, and meta-analyses indexed in MEDLINE (1984-06/2002) and the Cochrane Library (Issue 2, 2002) was conducted to show the effectiveness of GCS compared to nontreatment, other antithrombotic methods, or combined treatment. RESULTS: Of 48 surgeons 39 responded. Seven surgeons dismissed the use of GCS for thromboprophylaxis, 3 used GCS alone, 25 GCS in combination with heparin, and 4 used GCS only for patients at high risk. The review of the literature revealed the effectiveness of GCS in general and for abdominal surgical patients. Enhanced benefit is suggested when combining GCS with another intervention such as low-dose unfractionated heparin. Single application of GCS in orthopedic surgical or neurosurgical patients using venography showed no effect when compared to combined treatment of GCS and low molecular weight heparin. Trials with patients undergoing gynecological and urological surgeries are rare. There is a lack of trials investigating health-related quality of life and costs associated with the use of GCS. Complications are poorly reported. A determination as to the appropriate length of stockings is presently not possible. CONCLUSIONS: GCS should be integral part of DVT prophylaxis in surgical departments. Their ineffectiveness is likely in some surgical populations. PMID- 14740128 TI - [Graduated compression stockings in surgery -- optional or obligatory? Commentary invited by the editorship]. PMID- 14740129 TI - [Hilar cholangiocarcinoma -- results of en bloc resection of tumor and liver]. AB - The increasingly performed en bloc resection of liver and hilar tumor has contributed to the improvement of long-term survival in patients with hilar cholangiocarcinoma. Based on preoperative definition of operative strategy we tried to avoid any traumatization of the hilar region. Between September 1997 and September 2002, 82 patients with hilar cholangiocarcinoma were treated at our department. Three patients were excluded from any surgery. The resection rate was 75% (59 of 79); 79% (38 of 48) of en bloc resections of the hilar tumor and adjacent liver were formally curative. The hospital mortality was 7%. The 1- and 3-year survival rates of patients after explorative laparotomy, palliative and curative resection was 27 and 7%, 67 and 26%, 89 and 45% ( p<0.001), respectively. The 1- and 3-year survival rates of patients after en bloc resection were 78 and 49%, respectively. In patients with formally curative en bloc resection ( n=38), the 3-year survival rate was 63%; in patients with N0/R0 resection ( n=31) it was 71%. Lymph node involvement proved to be the only independent prognostic marker if patients who underwent hilar and en bloc resection were included in the multivariate analysis. The R situation was the only significant predictor for patients after en bloc resection. These data justify the extended diagnostic work-up and the principal liver resection in hilar cholangiocarcinoma. PMID- 14740130 TI - [Modification of the liver resection technique with radiofrequency coagulation]. AB - INTRODUCTION: Since the introduction of liver resection, intraoperative blood loss is considered to be a major risk factor for perioperative morbidity and mortality. Radiofrequency energy-an established technique for in situ ablation was used to facilitate liver resection by creating a nearly bloodless tissue plane. METHODS: After thorough manual and sonographic exploration of the liver, the planned and marked resection plane was coagulated by radiofrequency and then transected. Seven patients with liver metastases of colorectal carcinoma were operated on employing this technique. RESULTS: A nearly bloodless transection of the parenchyma could be achieved in all cases. Only two patients received blood transfusions: one due to a low preoperative hemoglobin and one due to extended additional abdominal surgery. Procedure-related complications included one case of a second-degree burn to the thigh and one case of postoperative bile leakage requiring hemihepatectomy. CONCLUSIONS: Radiofrequency-assisted resection offers a valuable additional option in liver surgery, especially in patients with an increased risk of intraoperative bleeding (cirrhosis, coagulopathy). PMID- 14740131 TI - [Angiosarcoma as a rare differential diagnosis in spontaneous rupture of the spleen]. AB - A splenectomy was performed in a 37-year-old woman because of spontaneous rupture of the spleen. Primary angiosarcoma of the spleen was diagnosed postoperatively based on histology. Primary angiosarcoma of the spleen is a very rare neoplasm with disastrous prognosis. Immunohistochemical and ultrastructural examinations are necessary to verify the diagnosis because the histopathological picture of the tumor is variable. Due to the small number of reported cases, there are no guidelines for adjuvant or palliative therapy, and up to today all adjuvant means of radiotherapy and chemotherapy have not improved the prognosis of the patients. PMID- 14740133 TI - [Does Kehr's sign derive from Hans Kehr? A critical commentary on its documentation?]. AB - Kehr's sign (pain in the left shoulder) is a well-known sign for a rupture of the spleen. After a thorough investigation into the body of research conducted by Hans Kehr, one must come to the conclusion that there is insufficient evidence that the term refers to his own work. The question of why the violent pain in the left shoulder during rupture of the spleen is named after the German gall bladder surgeon will remain a mystery of medical history. PMID- 14740132 TI - [Prolapsed malignant polyp of the stomach causing acute pancreatitis]. AB - Polyps of the stomach are rare lesions with an incidence between 1.7% and 3.9%. Until now, most cases were asymptomatic and found during endoscopic examination. Only three cases of prolapsing gastric polyps causing acute pancreatitis have been reported. Prolapsed polyps of the stomach should be included in the differential diagnosis of secondary pancreatitis. If endoscopic polypectomy is impossible, surgical treatment is necessary. In case early gastric cancer of the mucosa is detected in a polyp, limited surgery is possible. Otherwise, gastrectomy with lymphadenectomy of the D2 compartment is required PMID- 14740134 TI - [Concerning the article "Extirpation of the rectum in deep-seated rectal carcinoma" by F. Marusch et al, Chirurg (2003) 74:341-352]. PMID- 14740136 TI - Multicommutation as an environmentally friendly analytical tool in the hydride generation atomic fluorescence determination of tellurium in milk. AB - The aim of this study is to show the advantages of the emerging multicommutation methodology based on the use of solenoid valves for Te determination in milk by hydride generation atomic fluorescence spectrometry (HG-AFS). The delivery of a series of alternating sequential insertions of small volumes of samples and reagents gives rise to new hydrodynamic processes and exciting analytical potentials by controlling the time of flow through the on/off-switched solenoid valves. This drastically reduces the reagent consumption by a factor of 4 and the generation of effluents (590 mL h(-1) instead of 750 mL h(-1) generated by the continuous-mode measurement) and also provides an improvement in the laboratory productivity by an increase of the sample throughput (85 h(-1) compared to 20 h( 1) found in the continuous mode). So, multicommutation is an environmentally and economically sustainable alternative to the methodology based on continuous measurements. The multicommutation-based method developed was applied to tellurium determination in commercially available milk samples; a calibration range of 0.0-0.5 ng mL(-1) and a detection limit of 0.20 ng L(-1) with average relative standard deviation of 2.1% were found. Comparable results were obtained for a series of samples using both continuous and multicommutation HG-AFS modes. PMID- 14740137 TI - Characterisation of selected drugs with nitrogen-containing saturated ring structures by use of electrospray ionisation with ion-trap mass spectrometry. AB - The electrospray ionisation-ion-trap mass spectrometry (ESI-MS(n)) of selected drugs with nitrogen-containing saturated ring structures has been investigated. Sequential product-ion fragmentation experiments (MS(n)) have been performed to elucidate degradation pathways for the [M+H](+) ions and their predominant fragment ions. These MS(n) experiments result in characteristic fragmentations in which functional groups are generally cleaved from the ring systems as neutral molecules such as H(2)O, amines, alkenes, esters, carboxylic acids, etc. When such a nitrogen-containing drug molecule also contains a functional group, such as an ester, that on liberation as a neutral molecule has a significantly lower Delta H(f) degrees value than that of the corresponding amine then the former is preferentially liberated. Furthermore, when an aromatic entity is present in these drug molecules together with the nitrogen-containing saturated ring structure fragmentation of the latter ring occurs with the former, predictably, being resistant to fragmentation. The structures of fragment ions proposed for ESI-MS(n) can be supported by electrospray ionisation-quadrupole time-of-flight mass spectrometry (ESI-QTOFMS). The data presented in this paper therefore provide useful information on the structure of these heterocyclic compounds which could be used to characterise unknown drug compounds isolated from natural sources, for example. PMID- 14740139 TI - Characterization of honey amino acid profiles using high-pressure liquid chromatography to control authenticity. AB - Amino acid analysis of honey by high-performance liquid chromatography (HPLC) was used first to discriminate different botanical origins and then to combat adulteration. Pure honeys of seven selected floral varieties were examined. A principal component analysis (PCA) was carried out on the results after selection of the most discriminating parameters. Lavender honeys were thus perfectly characterized, but complete satisfaction was not obtained with the six other varieties. This method (analysis by HPLC and statistical processing by PCA) enabled us to detect the addition of sugar syrup to rape and fir honeys. PMID- 14740141 TI - Continuous ultrasound-assisted extraction coupled to a flow injection-flame atomic absorption spectrometric system for calcium determination in seafood samples. AB - Calcium was extracted on-line from solid seafood samples by a simple and rapid continuous ultrasound-assisted extraction system. This system is connected to a flow injection manifold, which allows the on-line flame atomic absorption spectrometric determination of calcium. This method enables the analysis of solid samples avoiding time-consuming traditional sample preparation methods and their inherent errors. The on-line manifold for calcium determination is the simplest possible, because a volume of 250 microL of acid extract is injected into an ultrapure water carrier stream. The acid extract was diluted on-line with lanthanum, which also acts as masking agent in order to avoid chemical interferences. The continuous monitoring of the calcium signal was accomplished by flame atomic absorption spectrometry. A Plackett-Burman experimental design was used for the optimisation of the continuous leaching procedure. The method allowed a total sampling frequency of 40 samples per hour, with a relative standard deviation for the complete procedure of 0.9% (for a sample containing 3414.35 mg/kg calcium (dry mass)). The limit of detection was found to be 44.4 mg/kg (dry mass) for 5 mg of sample. The analytical procedure was applied to real seafood samples. PMID- 14740142 TI - Use of a simplified generalized standard additions method for the analysis of cement, gypsum and basic slag by slurry nebulization ICP-OES. AB - The simplified generalized standard additions method (GSAM) was investigated as an alternative method for the ICP-OES analysis of solid materials, introduced into the plasma in the form of slurries. The method is an expansion of the conventional standard additions method. It is based on the principle of varying both the sample mass and the amount of standard solution added. The relationship between the sample mass, standard solution added and signal intensity is assumed to be linear. Concentration of the analyte can be found either geometrically from the slope of the two-dimensional response plane in a three-dimensional space or mathematically from the ratio of the parameters estimated by multiple linear regression. The analysis of a series of certified reference materials (CRMs) (cement CRM-BCS No 353, gypsum CRM-Gyp A and basic slag CRM No 382/I) introduced into the plasma in the form of slurry is described. The slurries contained glycerol and hydrochloric acid and were placed in an ultrasonic bath to ensure good dispersion. "Table curve 3D" software was used to fit the data. Results obtained showed that the method could be successfully applied to the analysis of cement, gypsum and slag samples, without the need to dissolve them. In this way, we could avoid the use of hazardous chemicals (concentrated acids), incomplete dissolution and loss of some volatiles. The application of the simplified GSAM for the analysis did not require a CRM with similar chemical and mineralogical properties for the calibration of the instrument. PMID- 14740143 TI - Direct observation of reconstruction induced changes of surface stress for Sb on Si(111). AB - With the combination of high resolution low energy electron diffraction and a bending sample technique we have simultaneously studied surface stress and surface structure during adsorption and desorption of antimony on the Si(111) surface. During desorption, several surface reconstructions with significantly different effects on the stress signal evolve. The surface stress of all observed structures has been obtained. PMID- 14740144 TI - Shipboard analysis of picomolar levels of manganese in seawater by chelating resin concentration and chemiluminescence detection. AB - A new shipboard analytical method for determining picomolar levels of manganese in seawater has been developed. The method is based on a combination of chelating resin column extraction and improved chemiluminescence (CL) detection in a closed flow system. In this method, manganese in sample solution is selectively collected on newly-developed iminodiacetate-immobilized chelating resin, and then eluted with acidic solution containing hydrogen peroxide. The resulting eluent is mixed with luminol solution and aqueous ammonia after removal of iron ions by a chelating resin column, and then the mixture is introduced into the CL cell. The manganese concentration is obtained from the CL intensity. The detection limit (3SD) of manganese is 5 pmol L(-1) from 9 mL of seawater sample. The method was applied to seawater samples collected at the Okinawa Trough. PMID- 14740146 TI - Effects of pregnanolone alone and in combination with other positive GABAA modulators on complex behavior in rats. AB - RATIONALE: Although positive modulators of gamma-aminobutyric acid(A) (GABA(A)) receptors generally produce similar behavioral effects, regardless of which modulatory site on the GABA(A) receptor complex mediates these effects, some differences have been observed between the effects of neuroactive steroids and those of other positive GABA(A) modulators. OBJECTIVE: The current study was designed to compare the behavioral effects of a neuroactive steroid to those of other positive GABA(A) modulators. METHODS: Rats responded under a multiple schedule of repeated acquisition and performance of response chains, with responding maintained under a second-order fixed-ratio 2 schedule of food presentation. RESULTS: Pregnanolone, flunitrazepam, pentobarbital and ketamine, an antagonist at NMDA receptors, dose-dependently decreased response rates and increased the percentage of errors in both components of the multiple schedule. Although the rate-decreasing and error-increasing effects of pregnanolone, pentobarbital and ketamine were quantitatively similar to each other, flunitrazepam was less effective in decreasing response rates and more effective in increasing errors than the other three drugs. A dose of 3.2 mg/kg pregnanolone potentiated the effects of flunitrazepam and pentobarbital, producing 2- to 3 fold shifts to the left in the dose-effect curves. In contrast, pregnanolone did not alter the ketamine dose-effect curves. CONCLUSIONS: The disruptive effects of the neuroactive steroid pregnanolone are qualitatively similar to those of other positive GABA(A) modulators as well as ketamine; however, the potentiation of the effects of flunitrazepam and pentobarbital, and not ketamine, emphasizes the importance of GABA(A) receptors in the behavioral effects of pregnanolone. PMID- 14740147 TI - A comparative analysis of the potential of cannabinoids and ondansetron to suppress cisplatin-induced emesis in the Suncus murinus (house musk shrew). AB - RATIONALE: The 5-HT3 antagonist, ondansetron (OND), and the cannabinoid, delta9 tetrahydrocannabinol (delta9-THC), have been shown to interfere with emesis; however, their relative and/or combined effectiveness in suppressing vomiting produced by the chemotherapeutic agent, cisplatin, is unknown. OBJECTIVE: To evaluate the potential of: 1) a broad range of doses of delta9-THC and OND to prevent cisplatin-induced vomiting and retching in the Suncus murinus (house musk shrew), 2) combined treatment with ineffective individual doses of delta9-THC and OND to prevent cisplatin-induced vomiting and retching, 3) the CB1 receptor antagonist, SR141716, to reverse the antiemetic effects of OND, and 4) cannabidiol (CBD), the principal non-psychoactive component of marijuana, to reverse cisplatin-induced vomiting in the shrew. METHODS: Shrews were injected with various doses of OND (0.02-6.0 mg/kg), delta9-THC (1.25-10 mg/kg) and a combination of ineffective doses of each (0.02 mg/kg OND+1.25 mg/kg delta9-THC) prior to being injected with cisplatin (20 mg/kg) which induces vomiting. Shrews were also injected with CBD (5 mg/kg and 40 mg/kg) prior to an injection of cisplatin. RESULTS: OND and delta9-THC both dose-dependently suppressed cisplatin induced vomiting and retching. Furthermore, a combined pretreatment of doses of the two drugs that were ineffective alone completely suppressed vomiting and retching. CBD produced a biphasic effect, suppressing vomiting at 5 mg/kg and potentiating it at 40 mg/kg. CONCLUSIONS: A low dose of the non-intoxicating cannabinoid CBD may be an effective anti-emetic treatment and combined doses of OND and delta9-THC that are ineffective alone suppresses cisplatin-induced emetic reactions in shrews. PMID- 14740148 TI - Nantenine: an antagonist of the behavioral and physiological effects of MDMA in mice. AB - RATIONALE: No selective antagonists for the effects of MDMA have yet been identified. The structurally-similar, naturally-occurring plant alkaloid nantenine (9,10-methylenedioxy-1,2 dimethoxyaporphine) may represent such a compound. OBJECTIVES: To investigate the capacity of nantenine to block and/or reverse MDMA-induced hyperthermia, lethality, locomotor stimulation, and head twitches in mice, and to compare these actions with those of the selective alpha1 antagonist prazosin and the selective 5-HT2A antagonist M100907. METHODS: Pretreatments of either 10 mg/kg nantenine or 1 mg/kg prazosin were administered 15 min before 32 mg/kg MDMA; core temperature and locomotor stimulation were then monitored via radiotelemetry for at least 3 h. In further hyperthermia studies, 32 mg/kg MDMA was administered first and temperature was allowed to rise for 30 min; 10 mg/kg nantenine, 1 mg/kg prazosin, or 1 mg/kg M100907 was then administered in an attempt to reverse MDMA-induced hyperthermia. In lethality assays, percent lethality was quantified 2 h after MDMA injection in two distinct housing conditions, one or 12 mice per cage, with or without 15 min pretreatments of 10 mg/kg nantenine or 1 mg/kg prazosin. Drug elicited head twitches were quantified for 10 min following administration of either MDMA enantiomer, with and without pretreatments of 1 mg/kg nantenine, 0.1 mg/kg prazosin, or 0.001 mg/kg M100907. RESULTS: Nantenine blocked and rapidly reversed MDMA-induced hyperthermia, attenuated lethality in both housing conditions, and reduced MDMA induced locomotor stimulation and head twitches in mice. Prazosin blocked, but did not reverse, MDMA-induced hyperthermia, attenuated lethality (more effectively in singly-housed animals), and reduced MDMA-induced locomotor stimulation and head twitches. M100907 did not reverse MDMA-induced hyperthermia, but effectively blocked drug-elicited head twitches. CONCLUSIONS: Nantenine functions as an effective antagonist against a wide range of MDMA-induced effects in mice. The antagonist actions of this compound at serotonin and adrenergic receptors may be differentially implicated across endpoints. PMID- 14740149 TI - Fluoxetine, but not sertraline or citalopram, potentiates the locomotor stimulant effect of cocaine: possible pharmacokinetic effects. AB - RATIONALE: The selective serotonin reuptake inhibitor (SSRI) fluoxetine enhances some of the behavioural effects of cocaine, including locomotor stimulation. While this effect has often been interpreted as evidence for a serotonergic component to the behavioural effects of cocaine, direct evidence for this hypothesis is lacking. One alternative explanation is that fluoxetine, by inhibiting cytochrome P450 (CYP) enzymes, interferes with the metabolism of cocaine. OBJECTIVES: These experiments were undertaken to: 1) compare the effects of fluoxetine with those of two other SSRIs, sertraline and citalopram, on cocaine-induced locomotor activity, 2) examine the effects of fluoxetine on cocaine-stimulated locomotion in rats depleted of serotonin (5-hydroxytryptamine; 5-HT), and 3) determine the effect of fluoxetine on cocaine levels in the brain. METHODS: Locomotor activity was measured, using photocell based activity monitors, in rats habituated to those monitors. Depletion of 5-HT was achieved by injecting 5,7-dihydroxytryptamine (5,7-DHT) into the dorsal and median raphe nuclei. Cocaine levels in whole brain were measured using high-performance liquid chromatography with ultraviolet detection. RESULTS: In experiment 1, 5 mg/kg fluoxetine enhanced the ability of 10 and 15 mg/kg cocaine to increase locomotor activity. Neither citalopram nor sertraline (5 and 10 mg/kg) altered the stimulant effect of 10 mg/kg cocaine. Experiment 2 showed that this effect of fluoxetine was also apparent in rats with large and widespread depletion of brain 5-HT levels. The 5-HT depletion also failed to alter the response to cocaine itself. In experiment 3, brain levels of cocaine were elevated in rats pretreated with fluoxetine compared with rats that received cocaine alone. CONCLUSION: Fluoxetine enhanced the ability of cocaine to increase locomotor activity. This effect appears not to depend upon increasing 5-HT function since fluoxetine was also effective in rats with substantial 5-HT depletions, and two other SSRIs did not alter the effects of cocaine. Fluoxetine-treated rats had higher brain levels of cocaine than rats treated with cocaine alone. This effect suggests that fluoxetine slows the metabolism of cocaine, perhaps by inhibition of CYP enzymes involved in metabolizing cocaine. The results also indicate that 5-HT reuptake inhibition may not play a prominent role in mediating the stimulant effects of cocaine. PMID- 14740150 TI - Age-dependent effects of nicotine on locomotor activity and conditioned place preference in rats. AB - RATIONALE: Most adult smokers start smoking during their adolescence. This adolescent initiation may be due to multiple factors, but little evidence is available regarding whether their brains are differentially sensitive to the addictive effects of nicotine during adolescence. OBJECTIVE: To test the hypothesis that adolescents are more sensitive than adults to nicotine's rewarding actions. METHODS: An unbiased, counterbalanced, place-conditioning procedure was used to examine drug-induced reward and locomotor activity. Early adolescent (postnatal day 28), late adolescent (P38) and adult (P90) rats received either saline or nicotine (0.125, 0.25 or 0.5 mg/kg, s.c.) and were tested for place conditioning. RESULTS: During early adolescence, a single nicotine injection (0.5 mg/kg) induced significant conditioned place preference (CPP). In contrast, during late adolescence or adulthood, nicotine did not induce CPP after either one or four conditioning trials. Initial locomotor responses to acute nicotine administration during the first conditioning trial also differed with age, with no effect at P28, but substantial inhibitory responses at all doses studied (0.125-0.5 mg/kg) at later ages. Although not differing in their initial locomotor response to nicotine, there was a significantly greater tolerance/sensitization during the second and subsequent drug exposures in late adolescents than in adults. CONCLUSIONS: These findings provide evidence that adolescent brain is differentially sensitive to both the acute and repeated effects of nicotine relative to adult brain. Furthermore, there are significant differences in nicotine sensitivity between early and late phases of adolescence. PMID- 14740151 TI - Can geometry-based parameters from pQCT and material parameters from quantitative ultrasound (QUS) improve the prediction of radial bone strength over that by bone mass (DXA)? AB - The diagnosis of osteoporosis is generally based on the assessment of bone mineral content with dual X-ray absorptiometry (DXA) but does not account for the spatial distribution and inherent material properties of the tissue. Peripheral quantitative computed tomography (pQCT) permits one to measure the compartment specific density and geometry-based parameters of cortical bone. Quantitative ultrasound (QUS) parameters are associated with material properties of cortical bone. The purpose of this study was to test the hypothesis that pQCT and cortical QUS provide additional information to DXA in predicting structural strength of the distal radius. The intact right arm and the isolated left radius were harvested from 70 formalin-fixed cadavers (age 79+/-11 years). The bone mineral content (BMC) was assessed with DXA at the radial metaphysis and shaft. pQCT was also used at the metaphysis and the shaft, while QUS was employed only at the shaft. The failure loads of the radius were assessed by use of a 3-point bending test (isolated radius) and a complex fall simulation (intact arm). The BMC (DXA) displayed a correlation of r=0.96 with the failure moments in 3-point bending ( P<0.001). The correlation between failure load and geometry-based parameters (pQCT) ranged from r=0.85 to r=0.96 and was r=0.64 for the speed of sound (QUS) ( P <0.001). Cortical thickness (pQCT) improved the prediction marginally (r=0.964) in combination with DXA. For the fall simulation, the correlation coefficients were r=0.76 for BMC (DXA) of the shaft, r=0.83 for metaphyseal bone content (pQCT), r=0.55 for QUS, and ranged from r=0.59 to r=0.74 for geometry-based parameters at the shaft (pQCT). pQCT and QUS parameters provided no significant improvement versus DXA alone. Measurement of bone mass by DXA or pQCT thus appears to be sufficient as a surrogate of mechanical strength and fracture risk of the distal radius. PMID- 14740154 TI - Tibial plateau fracture following anterior cruciate ligament reconstruction. AB - A case is presented of a tibial plateau fracture after previous anterior cruciate ligament reconstruction using patellar tendon autograft. The tibial plateau fracture occurred through the transosseous tibial tunnel and followed a torsional injury to the involved extremity. The stress riser effect of the transosseous tibial tunnel and the anatomic location of the cortical defect probably facilitated development of the fracture. Minimally invasive fixation of the fracture was effective in preserving knee stability without need for revision anterior cruciate ligament reconstruction. PMID- 14740155 TI - BTB ACL reconstruction: femoral nerve block has no advantage over intraarticular local anaesthetic infiltration. AB - Fifty patients were randomly recruited to receive either femoral nerve block (0.375% Bupivacaine) or an intraarticular local anaesthetic injection for pain control for arthroscopically-assisted ACL reconstruction. Both groups were evenly matched for age ( t-test p >0.05). Tourniquet time did not differ significantly between the groups ( t-test p=0.24). The VAS pain levels were not significantly different at 4 h and the first morning postoperatively in both groups. Femoral block (Median VAS: 20 & 18.5) did not confer a significant advantage (Mann Whitney U test p =0.36, 0.67) over intraarticular injection of bupivacaine (Median VAS: 18 & 20). There was no correlation between tourniquet time and postoperative pain ( r=0.19, 0.08). All patients but one were discharged home on the first postoperative morning. Our study demonstrates that pain levels can be sufficiently controlled by intraarticular infiltration of bupivacaine coupled with oral analgesia. The level of pain relief achieved could allow this procedure to be performed in a day surgery setting. PMID- 14740153 TI - Efficacy of alphacalcidol and calcitriol in primary and corticosteroid-induced osteoporosis: a meta-analysis of their effects on bone mineral density and fracture rate. AB - Vitamin D metabolites alphacalcidol and calcitriol (D-hormones) have been investigated for two decades, but few and conflicting results are available from high-quality randomized controlled trials. Our objectives were to provide an evidence-based update quantitatively summarizing their efficacy on bone mineral density (BMD) and fracture rate. We performed a systematic research of any randomized controlled trial containing relevant data, peer review, data extraction and quality scoring blinded for authors and data sources, and comprehensive meta-analyses of the relevant data. Inclusion criteria were: randomized controlled study, calcitriol or alphacalcidol, BMD or fractures in healthy/osteopenic/osteoporotic patients exposed or not to corticosteroids (CS). Analyses were performed in a conservative fashion using professional dedicated softwares and stratified by outcome, target patients, study quality, and control group type. Results were expressed as effect size (ES) for bone loss or relative risk (RR) for fracture while allocated to D-hormones vs control. Publication bias and robustness were investigated. Of the trials that were retrieved and subsequently reviewed, 17 papers fitted the inclusion criteria and were assessed. Quality scores ranged from 20 to 100%, the mean (standard deviation) being 72 (22)%. Calcitriol and alphacalcidol were found to have the same efficacy on all outcomes at p>0.13. We globally assessed D-hormones effects in preventing bone loss in patients not exposed to CS, and found positive effect: ES=0.39 ( p<0.001). For lumbar spine, this particular effect was 0.43 ( p<0.001). D hormones significantly reduced the overall fracture rates: RR=0.52 (0.46; 0.59) and both vertebral and non-vertebral fractures: RR=0.53 (0.47; 0.60) and RR=0.34 (0.16; 0.71), respectively. No statistical difference in response was observed between results from studies on healthy and osteoporotic patients or depending on the fact that controls were allowed to calcium supplementation. Treatment with D hormones was evaluated for maintaining spinal bone mass in five trials of patients with CS-induced osteoporosis, and provided ES=0.43 at p<0.001. Only two studies specifically addressed the effects of calcitriol on spinal fracture rate. None of them provided significant results, and the global RR did not reach the significance level as well: RR=0.33 (0.07; 1.51). Our data demonstrated efficacy for DH on bone loss and fracture prevention in patients not exposed to CS and on bone loss in patients exposed to CS, in the light of the most reliable scientific evidence. Their efficacy in reducing the number of fractures in patients exposed to CS remains to be determined. PMID- 14740156 TI - Short-term outcome of critically ill patients with severe acute respiratory syndrome. AB - OBJECTIVE: To document the outcome and determine prognostic factors for patients with severe acute respiratory syndrome who require admission to an intensive care unit. DESIGN: Observational cohort study involving retrospective analysis of demographic, clinical, laboratory and radiological data. SETTING: Adult intensive care unit in a tertiary referral university hospital involved in a major outbreak of severe acute respiratory syndrome (SARS). PATIENTS: The first 54 patients admitted with SARS to an intensive care unit (ICU). All were treated with corticosteroids, ribavirin, broad spectrum antimicrobials and supportive therapy. INTERVENTIONS: None. MEASUREMENTS AND RESULTS: All patients were admitted for respiratory failure. The median APACHE II score was 11 (interquartile range 8 13). At 28 days 34 patients (63%; 95% CI 49.6-74.6) were alive and not mechanically ventilated. Six patients were alive but ventilated (11.3%; 95% confidence interval 5.3-22.6) and 14 had died (25.9%; CI 16.1-38.9). Seven of 27 ventilated patients developed evidence of barotrauma (25.9%; 95% CI 13.2-44.7). Median maximal multiple-organ dysfunction score was 5 (interquartile range 3.3 9). Median maximal respiratory dysfunction score was 3 (interquartile range 3-4). Increased age, severity of illness, lymphocyte count, decreased steroid dose, positive fluid balance, chronic disease or immunosuppression and nosocomial sepsis were associated with poor outcome on univariate analysis. Poor outcome was defined as death or need for mechanical ventilation at 28 days after ICU admission. CONCLUSIONS: Mortality amongst critically ill patients with SARS is high. It causes predominantly severe respiratory failure, with little other organ failure, and a high incidence of barotrauma amongst those requiring mechanical ventilation. PMID- 14740157 TI - Infectious and inflammatory dissemination are affected by ventilation strategy in rats with unilateral pneumonia. AB - OBJECTIVE: To evaluate the effect of V(T) reduction and alveolar recruitment on systemic and contralateral dissemination of bacteria and inflammation during right-side pneumonia. DESIGN: Interventional animal study. SETTING. University hospital research laboratory. SUBJECTS: A total of 54 male Wistar rats. INTERVENTIONS: One day after right lung instillation of 1.4x10(7) Pseudomonas aeruginosa, rats were left unventilated or ventilated for 2 h at low V(T) (6 ml/kg) with different strategies of alveolar recruitment: no PEEP, 8 cm H(2)O PEEP, 8 cm H(2)O PEEP in a left lateral position, 3 cm H(2)O PEEP with partial liquid ventilation, or high V(T) (set such as end-inspiratory pressure was 30 cm H(2)O) without PEEP (ZEEP). After ventilation the lungs, spleen and liver were cultivated for bacterial counts. Global bacterial dissemination was scored considering the percentage of positive spleen, liver and left lung cultures. TNF alpha was assayed in plasma before and after mechanical ventilation. MEASUREMENTS AND RESULTS: All rats had right-side pneumonia with similar bacterial counts. All mechanical ventilation strategies, with the exception of low V(T)-PEEP 8, promoted contralateral lung dissemination. Overall bacterial dissemination was less in non-ventilated controls (22%) and low V(T)-PEEP 8 (22%) than in high V(T) ZEEP (67%), low V(T)-PEEP 8 in left lateral position (59%) and low V(T)-ZEEP (56%) ( p<0.05). Partial liquid ventilation prevented systemic bacterial translocation, but at the expense of contralateral bacterial seeding. Plasma TNF alpha concentration increased significantly after mechanical ventilation with no PEEP at both high and low V(T). CONCLUSIONS: Our results suggest that PEEP might reduce the risk of ventilation-induced bacterial and inflammatory mediator dissemination during pneumonia. PMID- 14740158 TI - Long-chain CoA esters activate human pancreatic beta-cell KATP channels: potential role in Type 2 diabetes. AB - AIMS/HYPOTHESIS: The ATP-regulated potassium (KATP) channel in the pancreatic beta cell couples the metabolic state to electrical activity. The primary regulator of the KATP channel is generally accepted to be changes in ATP/ADP ratio, where ATP inhibits and ADP activates channel activity. Recently, we showed that long-chain CoA (LC-CoA) esters form a new class of potent KATP channel activators in rodents, as studied in inside-out patches. METHODS: In this study we have investigated the effects of LC-CoA esters in human pancreatic beta cells using the inside-out and whole-cell configurations of the patch clamp technique. RESULTS: Human KATP channels were potently activated by acyl-CoA esters with a chain length exceeding 12 carbons. Activation by LC-CoA esters did not require the presence of Mg2+ or adenine nucleotides. A detailed characterization of the concentration-dependent relationship showed an EC50 of 0.7+/-0.1 micromol/l. Furthermore, in the presence of an ATP/ADP ratio of 10 (1.1 mmol/l total adenine nucleotides), whole-cell KATP channel currents increased approximately six-fold following addition of 1 micro mol/l LC-CoA ester. The presence of 1 micro mol/l LC-CoA in the recording pipette solution increased beta-cell input conductance, from 0.5+/-0.2 nS to 2.5+/-1.3 nS. CONCLUSION/INTERPRETATION: Taken together, these results show that LC-CoA esters are potent activators of the KATP channel in human pancreatic beta cells. The fact that LC-CoA esters also stimulate KATP channel activity recorded in the whole-cell configuration, points to the ability of these compounds to have an important modulatory role of human beta-cell electrical activity under both physiological and pathophysiological conditions. PMID- 14740159 TI - Common genetic polymorphisms in the promoter of resistin gene are major determinants of plasma resistin concentrations in humans. AB - AIMS/HYPOTHESIS: Resistin is thought to be an important link between obesity and insulin resistance. It has been suggested that genetic polymorphism in the promoter of resistin gene is a determinant of resistin mRNA expression and possibly associated with obesity and insulin resistance. In this study, we investigated the association between the genotype of resistin promoter and its plasma concentrations. METHODS: We examined g.-537A>C and g.-420C>G polymorphisms in the resistin promoter and measured plasma resistin concentrations in Korean subjects with or without Type 2 diabetes. We also did haplotype-based promoter activity assays and the gel electrophoretic mobility shift assay. RESULTS: The 420G and the -537A alleles, which were in linkage disequilibrium, were associated with higher plasma resistin concentrations. Individuals with haplotype A-G (-537A and -420G) had significantly higher plasma resistin concentrations than the others. Haplotype A-G had modestly increased promoter activity compared to the other haplotypes. Electrophoretic mobility shift assay showed that the -420G allele is specific for binding of nuclear proteins from adipocytes and monocytes. However, none of the two polymorphisms were associated with Type 2 diabetes or obesity in our study subjects. CONCLUSIONS/INTERPRETATION: Polymorphisms in the promoter of resistin gene are major determinants of plasma resistin concentrations in humans. PMID- 14740160 TI - Extracorporeal shock wave lithotripsy in children: experience using a mpl-9000 lithotriptor. AB - Our objectives were to assess the value of extracorporeal shock wave lithotripsy (ESWL) in treating pediatric urolithiasis, and to determine the factors that may affect treatment success. Between January 1993 and August 2002, 129 children with upper urinary tract calculi (134 renoureteral units) were treated using a Dornier MPL-9000 lithotriptor. The series consisted of 77 boys and 52 girls with an age range from 20 months to 14 years (average age: 8.7 years). All ESWL procedures took place under general anaesthesia or sedation with ketamin or fentanyl. Under ultrasonic or fluoroscopic guidance, children were treated with a maximum 2,550 shocks at an average of 19.5 kV. Success was defined as the lack of any visible stone fragments on post-treatment radiological evaluation. The patients were assessed 3 months after ESWL treatment and the results were compared using chi(2) tests to detect factors that might be associated with treatment success. There were 105 renal, 20 ureteral, four bilateral renal and one unilateral renal plus contralateral ureteral calculi. The mean sizes were 15.7 mm for pelvic, 17.8 mm for renal and 10.2 mm for ureteral stones. One or two lithotripsy sessions were sufficient in most cases (71.6%). In 15 (11.6%) patients, double J stents introduced before lithotripsy were left indwelling until all stone fragments were voided. Overall success rates were 89.5% for pelvic, 85.5% for renal and 75% for ureteral stones. Complications such as urinary tract infection, Steinstrasse and small subcapsular hematoma occurred in 19 (14.7%) patients. The only significant factor associated with the stone-free rate was the diameter of the stone ( P=0.022). This study confirmed that the stone-free rate is significantly influenced by stone size. Because children with stone disease are at risk for a longer period than adults, their cumulative likelihood of stone recurrences may be higher. Thus, we agree with other authorities that minimally invasive treatment, such as ESWL, is mandatory in children with urolithiasis. PMID- 14740161 TI - Lymphangiosis carcinomatosa of the liver deriving from gastric carcinoma with a unique branching calcification. PMID- 14740162 TI - Mediastinal sarcoidosis induced by high-dose alpha-2-interferon therapy in a patient with malignant melanoma. PMID- 14740164 TI - Exophytic pilocytic astrocytoma of the brain stem in an adult with encasement of the caudal cranial nerve complex (IX-XII): presurgical anatomical neuroimaging using MRI. AB - We describe a rare case of adult pilocytic astrocytoma in which exophytic growth from the brain stem presented as a right cerebellopontine angle mass. An initial MRI examination using T2- and T1-weighted images without and with contrast suggested the diagnosis of schwannoma. Subsequent use of 3D CISS (three dimensional constructive interference in steady state) and T1-weighted contrast enhanced 3D MP-RAGE (three-dimensional magnetization prepared rapid acquisition gradient echo) sequences led to the diagnosis of an exophytic brain stem tumor, documented the precise relationships of the tumor to cranial nerve VIII, revealed encasement of cranial nerves IX-XII (later confirmed intraoperatively), and provided the proper basis for planning surgical management. PMID- 14740163 TI - Assessment of pulmonary hypertension by CT and MR imaging. AB - In the recent World Health Organization (WHO) classification the group of pulmonary arterial hypertension (PH) comprises the classic primary pulmonary hypertension and several conditions with definite or very high risk factors to develop pulmonary arterial hypertension. Therapeutic advances drive the need for a comprehensive pre-therapeutic evaluation for optimal treatment. Furthermore, follow-up examinations need to be performed to monitor changes in disease status and response to therapy. Up to now, the diagnostic imaging work-up of PH comprises mainly echocardiography, invasive right heart catheterization and ventilation/perfusion scintigraphy. Due to technical advances helical computed tomography (CT) and magnetic resonance imaging (MRI) became more important in the evaluation and for differential diagnosis of pulmonary arterial hypertension. Both modalities are reviewed and recommendations for clinical use are given. PMID- 14740165 TI - Management of acute adverse reactions to contrast media. AB - When anaphylactoid and other severe adverse reactions to contrast media occur, prompt recognition and immediate treatment are essential. Simple guidelines for treatment have been requested by many radiologists, and therefore the Contrast Media Safety Committee has produced guidelines for treatment of acute adverse reactions to contrast media. The committee made an extensive review of the literature on treatment of adverse reactions to contrast media. Based on this, a report and guidelines were prepared. The resulting report was discussed at the 10th European Symposium on Urogenital Radiology in Uppsala. Sweden, September 2003. Guidelines for treatment of acute adverse reactions and a list of first line drugs and equipment that should be available in the room where contrast medium is given are provided. PMID- 14740166 TI - MRI of hyperacute stroke in the AChA territory. AB - The purpose of our study was to derive from the anatomical literature an easy-to use map of the brain areas supplied by the anterior choroidal artery (AChA) and to assess the correspondence between damage within the putative AChA areas and clinical symptoms. A thorough review of the literature led to the recognition of 16 anatomical areas which could be delineated on routine diffusion-weighted MR images. A database of 138 consecutive ischemic stroke patients examined with MRI less than 6 h after symptoms onset was thereafter processed in a retrospective way. Patients presenting with at least one damaged AChA area were selected so as to assess the prevalence of AChA infarction and the clinical correlates of the condition. Fifteen patients (11%) had at least one damaged AChA area. Only two of them had "pure" AChA-restricted infarction. Contralateral hemiparesis and contralateral hemianesthesia were best predicted by lesions within the tail of the caudate nucleus with a sensitivity of 87% and 83%, respectively. Homonymous hemianopsia best correlated with lesions within the posterior limb of the internal capsule and within the retrolenticular part of the internal capsule, with a sensitivity of 100% and a specificity of 70% for both areas. We concluded that the clinical-radiological correlations did not match the neurophysiological standards, thereby highlighting the limitation of this study, which involved a cohort of acute stroke patients recruited from clinical practice and investigated the clinical impact of these brain lesions, even when documented with the most sensitive imaging modality. PMID- 14740167 TI - Improvement of cotton fiber quality by transforming the acsA and acsB genes into Gossypium hirsutum L. by means of vacuum infiltration. AB - A novel method for the genetic transformation of cotton pollen by means of vacuum infiltration and Agrobacterium-mediated transformation is reported. The acsA and acsB genes, which are involved in cellulose synthesis in Acetobacter xylinum, were transferred into pollen grains of brown cotton with the aim of improving its fiber quality by incorporating useful prokaryotic features into the colored cotton plants. Transformation was carried out in cotton pollen-germinating medium, and transformation was mediated by vector pCAMBIA1301, which contains a reporter gene beta-glucuronidase (GUS), a selectable marker gene, hpt, for hygromycin resistance and the genes of interest, acsA and acsB. The integration and expression of acsA, acsB and GUS in the genome of transgenic plants were analyzed with Southern blot hybridization, PCR, histochemical GUS assay and Northern blot hybridization. We found that following pollination on the cotton stigma transformed pollen retained its capability of double-fertilization and that normal cotton seeds were produced in the cotton ovary. Of 1,039 seeds from 312 bolls pollinated with transformed pollen grains, 17 were able to germinate and grow into seedlings for more than 3 weeks in a nutrient medium containing 50 mg/l hygromycin; eight of these were transgenic plants integrated with acsA and acsB, yielding a 0.77% transformation rate. Fiber strength and length from the most positive transformants was 15% greater than those of the control (non transformed), a significant difference, as was cellulose content between the transformed and control plants. Our study suggests that transformation through vacuum infiltration and Agrobacterium mediated transformation can be an efficient way to introduce foreign genes into the cotton pollen grain and that cotton fiber quality can be improved with the incorporation of the prokaryotic genes acsA and acsB. PMID- 14740168 TI - A large-scale Agrobacterium-mediated transformation procedure with a strong positive-negative selection for gene targeting in rice (Oryza sativa L.). AB - A large-scale transformation procedure handling an adequate number of stable transformants with highly efficient positive-negative selection is a necessary prerequisite to successful gene targeting by homologous recombination, as the integration of a transgene by somatic homologous recombination in higher plants has been reported to be 10(-3) to 10(-5) compared with random integration by non homologous end joining. We established an efficient and large-scale Agrobacterium mediated rice transformation protocol that generated around 10(3) stable transformants routinely from 150 seeds and a strong positive-negative selection procedure that resulted in survivors at 10(-2) using the gene for diphtheria toxin A fragment as a negative marker. The established transformation procedure provides a basis for efficient gene targeting in rice. PMID- 14740169 TI - Varicella glomerulonephritis mimicking microscopic polyangiitis. AB - Varicella in childhood is usually a self-limiting illness with few complications. Varicella nephritis is an uncommon entity and seen mostly in immunocompromized individuals. We report a 14-year-old boy with juvenile rheumatoid arthritis who developed varicella nephritis and in whom the renal manifestations preceded the skin lesions by 1 week. This is an extremely unusual occurrence, and only one case has been described before. Such a presentation can mimic the clinical features of microscopic polyangiitis. PMID- 14740170 TI - Seizure associated with chloroquine therapy in a patient with systemic lupus erythematosus. PMID- 14740173 TI - Iron status in 358 apparently healthy 80-year-old Danish men and women: relation to food composition and dietary and supplemental iron intake. AB - In Denmark, the intake of dietary iron has decreased since 1987, when the mandatory iron fortification of flour (30 mg carbonyl iron/kg) was stopped. Since there have been no studies of iron status in elderly Danes after the abolishment of iron fortification, there is a need to assess actual iron status in the elderly population. The objective was to evaluate iron status and the relationship with food composition and dietary and supplemental iron intake in an elderly population in Copenhagen County. Participants in this health examination survey were 358 subjects (171 men, 187 women) 80 years of age from a 1914 cohort study. Blood samples included serum ferritin and hemoglobin (Hb). A dietary survey was performed in 232 subjects (120 men, 112 women) using a dietary history method. Median serum ferritin was 100 microg/l in men and 78 microg/l in women ( p<0.001). Ferritin concentrations <16 microg/l (i.e., depleted iron stores) were found in three men (2%) and in ten women (5%). Median Hb was 140 g/l in men and 131 g/l in women ( p<0.001). Three subjects (0.84%) had iron deficiency anemia (i.e., ferritin <13 microg/l and Hb <5th percentile for iron-replete subjects (121 g/l in men, 114 g/l in women). Ferritin concentrations >300 microg/l (i.e., iron overload) were found in 15 (9%) men and in 5 (3%) women. Median dietary iron intake was higher in men (8.7 mg/day) than in women (7.3 mg/day) ( p<0.001). Serum ferritin was positively correlated to dietary intake of iron, meat, and alcohol and to body mass index in men. Serum ferritin displayed a negative correlation to the consumption of tea. The use of vitamin-mineral supplements containing iron had no influence on iron status. Dietary intake of iron and/or the bioavailability of dietary iron were adequate to maintain a favorable iron status in 80-year-old subjects displaying a low prevalence of iron deficiency and a moderate prevalence of iron overload. PMID- 14740174 TI - Ii-Key/HER-2/neu MHC class-II antigenic epitope vaccine peptide for breast cancer. AB - PURPOSE: Cytotoxic T lymphocytes (CTL)- and T-helper cell-specific, and major histocompatibility complex (MHC) class-I and class-II peptides, respectively, of the HER-2/ neu protein, induce immune responses in patients. A major challenge in developing cancer peptide vaccines is breaking tolerance to tumor-associated antigens which are functionally self-proteins. An adequate CD4+ T-helper response is required for effective and lasting responses. METHODS: Stimulating anti-cancer CD4+ T cell responses by MHC class-II epitope peptides has been limited by their weak potency, at least compared with tight-binding MHC class-I epitope peptides. Previously, a potent T-cell response to a MHC class-II epitope was engineered by coupling the N-terminus of the pigeon cytochrome C [PGCC(95-104)] MHC class-II epitope to the C-terminus of an immunoregulatory segment of the Ii protein (hIi77 81, the Ii-Key peptide) through a polymethylene spacer. RESULTS: In vitro presentation of the MHC class-II epitope to a T hybridoma was enhanced greatly (>250 times). Now, an Ii-Key/HER-2/neu (777-789) MHC class-II epitope hybrid peptide stimulated lymphocytes from both a healthy donor and a patient with metastatic breast carcinoma. The in vitro primary stimulation with the hybrid peptide strongly activated IFN-gamma release, whereas the epitope-only peptide was weakly active. In fact, the hybrid stimulated IFN-gamma release as well as the wild-type peptide when augmented with IL-12; however, the hybrid was comparable to free peptide in stimulating IL-4 release. This pattern is consistent with preferential activation along a non-tolerogenic Th1 pathway. CONCLUSION: Such Ii-Key/MHC class-II epitope hybrid peptides have both diagnostic and therapeutic applications. PMID- 14740175 TI - A primer on cancer immunology and immunotherapy. AB - The role of immunity in cancer has been abundantly demonstrated in murine tumor models as well as in man. Induction of clinically effective antitumor immune responses, based on this information, in patients with cancer however, remains elusive. This is not because tumors lack recognizable antigens [in fact there is evidence that there are thousands of potential novel targets in each tumor cell] but rather due to the fact that the induction of responses is not adequate nor particularly well understood. Tumors seem to be rather effective at limiting immune responses. Many of the molecularly defined antigens that have been detected on tumor cells are derived from self-proteins and as such are subject to tolerizing mechanisms. Such tumors have also developed escape mechanisms capable of evading or suppressing immune responses. Understanding the role of dendritic cells during the effector phase of the immune response and the complex interactions of stromal, immune, and tumor cells in the tumor microenvironment represent the next challenges to be understood for tumor immunology. PMID- 14740176 TI - Dendritic cells are dysfunctional in patients with operable breast cancer. AB - BACKGROUND: Dendritic cells (DCs) play a crucial role in presenting antigens to T lymphocytes and inducing cytotoxic T cells. DCs have been studied in patients with breast cancer to define the factors leading to failure of an effective systemic and locoregional anticancer host response. METHODS: Purified DCs were obtained from peripheral blood (PB) and lymph nodes (LNs) of women with operable breast cancer, using immunomagnetic bead selection. The stimulatory capacity of DCs in the allogeneic mixed leukocyte reaction (MLR) and autologous T cell proliferation test (purified protein derivative (PPD) as stimulator), the expression of surface markers on DCs and the production of cytokines in vitro by DCs from patients with operable breast cancer and from healthy donors (controls) were studied. RESULTS: 70-75% purified DCs were isolated from PB and LNs. PBDCs and LNDCs from patients with operable breast cancer demonstrated a reduced capacity to stimulate in an MLR, compared with PBDCs from normal donors (p<0.01). Autologous T cell proliferation in patients had a decreased ability to respond to PPD, when compared with controls (p<0.01). However, T cells from patients responded as well as control T lymphocytes in the presence of control DCs. PBDCs and LNDCs from patients expressed low levels of HLA-DR and CD86, and induced decreased interleukin-12 (IL-12) secretion in vitro, compared with DCs from normal donors (p<0.01). CONCLUSION: These data suggest a defective DC function in patients with operable breast cancer. Switched-off DCs in patients with early breast cancer and decreased IL-12 production may be important factors for progressive tumour growth. PMID- 14740177 TI - Long-term prognostic value of exercise 99mTc-MIBI SPET myocardial perfusion imaging in patients after percutaneous coronary intervention. AB - The purpose of this study was to evaluate the long-term prognostic value of exercise technetium-99m methoxyisobutylisonitrile ((99m)Tc-MIBI) single-photon emission tomography (SPET) imaging in patients after percutaneous coronary intervention (PCI). Three hundred and eighteen consecutive post-PCI patients who underwent exercise and rest (99m)Tc-MIBI SPET myocardial perfusion imaging (MPI) were followed up for 38+/-27 months. Patients with early revascularisation (<3 months after MPI) were excluded. A semiquantitative visual analysis employing a 20-segment and four-point scoring system was used to define the summed stress score (SSS), summed rest score (SRS) and summed difference score (SDS). Death and non-fatal myocardial infarction (MI) were considered as hard events, and late revascularisation procedures (>/=3 months after MPI) as soft events. Fifty-one patients (16.0%) suffered from cardiac events during follow-up, including 1 (0.3%) death, 13 (4.1%) non-fatal MIs, 9 (2.8%) coronary artery bypass grafting procedures and 28 (8.8%) PCIs. According to the SPET results, patients were classified into three groups: patients with normal MPI (SSS=0, n=153), patients with irreversible defects (SDS<3 and SRS>1, n=100) and patients with reversible defects (SDS>/=3, n=65). The annual hard cardiac event rate in patients with reversible defects was 3.9%, which was significantly higher than that in patients with normal MPI (0.2%, chi(2)=7.71; P<0.01). The annual soft cardiac event rate in patients with reversible defects was 10.7%, which was significantly higher than that in patients with irreversible defects (2.5% chi(2)=17.69; P<0.001), and also significantly higher than that in patients with normal MPI (1.5%, chi(2)=33.89; P<0.001). In patients with normal and reversible defects, there was no significant difference in soft and hard cardiac event rates according to whether patients were symptomatic or asymptomatic ( P>0.05). However, the annual soft event rate in patients with irreversible defects and symptoms was 5.0%, which was higher than that of 0.6% in asymptomatic patients (chi(2)=6.11, P<0.05). Multivariate Cox analysis showed that SSS was the best independent predictor for hard cardiac events (chi(2)=12.70; P<0.001) and SDS was the strongest independent predictor for soft cardiac events (chi(2)=11.72; P<0.001). Post-PCI patients who have normal exercise (99m)Tc-MIBI SPET MPI have a good long term prognosis, while those with reversible defects are at a higher risk for future cardiac events, without correlation to the chest pain symptoms. However, symptomatic patients with irreversible defects have a higher risk for repeat revascularisation, but not for hard events, compared with asymptomatic patients. Exercise (99m)Tc-MIBI SPET MPI has important clinical value for risk stratification and management decision-making in post-PCI patients. PMID- 14740178 TI - Estimating the input function non-invasively for FDG-PET quantification with multiple linear regression analysis: simulation and verification with in vivo data. AB - A novel statistical method, namely Regression-Estimated Input Function (REIF), is proposed in this study for the purpose of non-invasive estimation of the input function for fluorine-18 2-fluoro-2-deoxy- d-glucose positron emission tomography (FDG-PET) quantitative analysis. We collected 44 patients who had undergone a blood sampling procedure during their FDG-PET scans. First, we generated tissue time-activity curves of the grey matter and the whole brain with a segmentation technique for every subject. Summations of different intervals of these two curves were used as a feature vector, which also included the net injection dose. Multiple linear regression analysis was then applied to find the correlation between the input function and the feature vector. After a simulation study with in vivo data, the data of 29 patients were applied to calculate the regression coefficients, which were then used to estimate the input functions of the other 15 subjects. Comparing the estimated input functions with the corresponding real input functions, the averaged error percentages of the area under the curve and the cerebral metabolic rate of glucose (CMRGlc) were 12.13+/-8.85 and 16.60+/ 9.61, respectively. Regression analysis of the CMRGlc values derived from the real and estimated input functions revealed a high correlation (r=0.91). No significant difference was found between the real CMRGlc and that derived from our regression-estimated input function (Student's t test, P>0.05). The proposed REIF method demonstrated good abilities for input function and CMRGlc estimation, and represents a reliable replacement for the blood sampling procedures in FDG PET quantification. PMID- 14740179 TI - Relation of coronary vasoreactivity and coronary calcification in asymptomatic subjects with a family history of premature coronary artery disease. AB - Electron-beam computed tomography (EBCT) allows non-invasive imaging of coronary calcification and has been promoted as a screening tool for coronary artery disease (CAD) in asymptomatic high-risk subjects. This study assessed the relation of coronary calcifications to alterations in coronary vascular reactivity by means of positron emission tomography (PET) in asymptomatic subjects with a familial history of premature CAD. Twenty-one subjects (mean age 51+/-10 years) underwent EBCT imaging for coronary calcifications expressed as the coronary calcium score (CCS according to Agatston) and rest/adenosine-stress nitrogen-13 ammonia PET with quantification of myocardial blood flow (MBF) and coronary flow reserve (CFR). The mean CCS was 237+/-256 (median 146, range 0 915). The CCS was <100 in eight subjects and >100 units in 13. As defined by age related thresholds, 15 subjects had an increased CCS (>75th percentile). Overall mean resting and stress MBF and CFR were 71+/-16 ml 100 g(-1) min(-1), 218+/-54 ml 100 g(-1) min(-1) and 3.20+/-0.77, respectively. Three subjects with CCS ranging from 114 to 451 units had an abnormal CFR (<2.5). There was no relation between CCS and resting or stress MBF or CFR ( r=0.17, 0.18 and 0.10, respectively). In asymptomatic subjects a pathological CCS was five times more prevalent than an abnormal CFR. The absence of any close relationship between CCS and CFR reflects the fact that quantitative myocardial perfusion imaging with PET characterises the dynamic process of vascular reactivity while EBCT is a measure of more stable calcified lesions in the arterial wall whose presence is closely related to age. PMID- 14740180 TI - Variation in scanning line source sensitivity: a significant source of error in simultaneous emission-transmission tomography. AB - Transmission measurement is recommended in order to accurately correct for attenuation in myocardial single-photon emission tomography (SPET) studies. It is important that transmission studies are artefact-free, otherwise the attenuation corrected SPET studies may also be affected. An assumption in transmission studies is that the measured transmission in air used as a reference scan is valid for any camera orientation. Variation in transmission source sensitivity (both source efficiency and detector sensitivity) with rotation negates this assumption and can produce errors that result in significant reconstructed artefacts. The aim of this study was to investigate the variation in transmission source sensitivity with a view to defining action thresholds for routine quality control tests. Transmission measurements in air were recorded on two commercial scanning line source installations for the 180 degrees arc normally used in myocardial SPET. Significant variation in transmission source sensitivity was observed on one system (exceeding 30%). Comparison was also made with the reference scan recorded at a different time at a fixed angular location. Both systems demonstrated measurable variation between transmission counts and the corresponding reference scan. A simulation study was undertaken using patient data to determine the influence of transmission sensitivity variation on reconstructed myocardial counts. To maintain reconstructed counts to within 15% of that obtained with artefact-free transmission data, the variation in transmission sensitivity with rotation needed to be within 5%. These results have necessitated the addition of quality control procedures and specific maintenance procedures to attempt rectification of the problem. Variation in transmission source sensitivity with rotation is a potential source of error in attenuation corrected SPET. Steps should be taken to stabilise transmission source mountings so as to minimise this potential source of error. PMID- 14740181 TI - Cyclic sciatica caused by infiltrative endometriosis: MRI findings. AB - Endometriosis, an important gynecological disorder of reproductive women, affects most commonly the ovaries and less frequently the gastrointestinal tract, chest, urinary tract, and soft tissues. Endometriosis classically appears on MRI as a mass with a large cystic component and variable signal intensities on T1- and T2 weighted images due to the presence of variable degradation of hemorrhagic products. Endometriosis in an atypical location, an infiltrative appearance and without cystic-hemorrhagic components has rarely been described. We report on a 33-year-old woman with cyclic sciatica due to histologically documented infiltrative endometriosis involving the area of the left sciatic notch. PMID- 14740182 TI - A solitary lesion of talus with mixed sclerotic and lytic changes: Rosai-Dorfman disease of 25 years' duration. AB - Rosai-Dorfman disease (sinus histiocytosis with massive lymphadenopathy) is an unusual form of histiocytic disorder predominantly affecting children and young adults. Bone lesions are infrequent. We describe a 63-year-old woman with a solitary lesion of the talus which was misdiagnosed for 25 years. The patient never had adenopathy or other organ involvement. The radiographic findings were a mixture of sclerotic and lytic changes, an extremely rare and unusual presentation of the disease. PMID- 14740183 TI - Whole spine MRI in the assessment of acute vertebral body trauma. AB - OBJECTIVE: To determine the incidence and types of multilevel vertebral body injury in association with acute spinal trauma as assessed by whole spine MRI. DESIGN AND PATIENTS: All acute admissions to a regional spinal injury unit had whole spine MRI carried out, to detect occult vertebral body injury. Two radiologists assessed 127 cases prospectively, over a period of 3 years. All cases had T2-weighted sagittal imaging of the whole spine (where possible using a T2-weighted fat-suppressed sequence), with T1-weighted imaging in both sagittal and axial planes covering the primary injury. The incidence of secondary injury (defined as either bone bruising, wedge compression fracture or burst fracture) was determined and defined by type, site and relationship to the primary injury. RESULTS: Seventy-seven per cent of cases had a secondary injury level. Of these, bone bruising was the commonest but often occurred in combination with secondary wedge compression fracture or burst fracture. MRI detected 27 non-contiguous wedge compression fractures and 16 non-contiguous burst fractures, giving an incidence of secondary level, non-contiguous fracture of approximately 34%. CONCLUSION: A higher frequency of secondary vertebral body injury may be defined by MRI than has been described in previous studies based on radiographic evaluation of the whole spine. Whole spine MRI in assessment for occult vertebral body fracture enables increased confidence in the conservative or surgical management of patients with severe spinal injury. PMID- 14740184 TI - Intracortical epidermoid cyst of the tibia. AB - Epidermoid cyst in a long bone is an extremely rare condition. The authors describe such a tumor located in the cortex of the tibia in a 21-year-old woman. She was successfully treated with curettage and autogenous bone grafting. PMID- 14740185 TI - Osteoid osteoma simulating an osteocartilaginous exostosis. AB - We describe a case of osteoid osteoma in the tibia of a 3-year-old patient who presented with a clinical and radiographic picture that suggested an exostosis. The formation of osteoid osteoma with a radiographic picture similar to that of osteophytes or exostosis has been previously documented only rarely. The authors hypothesize that the exostosis-like formation observed was actually the calcification of soft tissues that formed after the intense periosteal inflammatory reaction caused by the osteoid osteoma. As a result of its peculiar clinical and radiographic presentation, diagnosis of this lesion was delayed. Being located close to the medial growth plate of the tibia, it caused lengthening of the limb with a pronounced valgus deviation of the knee. An excisional biopsy provided histological evidence, clinical resolution and immediate pain relief, but incomplete resolution of the valgus deformity of the knee. PMID- 14740186 TI - Biotechnological production and applications of the omega-3 polyunsaturated fatty acid docosahexaenoic acid. AB - Docosahexaenoic acid (DHA) is a polyunsaturated fatty acid composed of 22 carbon atoms and six double bonds. Because the first double bond, as counted from the methyl terminus, is at position three, DHA belongs to the so-called omega-3 group. In recent years, DHA has attracted much attention because of its beneficial effect on human health. At present, fish oil is the major source of DHA, but alternatively it may be produced by use of microorganisms. Marine microorganisms may contain large quantities of DHA and are considered a potential source of this important fatty acid. Some of these organisms can be grown heterotrophically on organic substrates without light. These processes can be well controlled and DHA with constant quality can be produced all year round. This paper reviews recent advances in the biotechnological production of DHA by marine microorganisms. PMID- 14740187 TI - Studies on molecular mechanisms of Ginkgo biloba extract. AB - In the past decade, interest by the general public in the use of herbal dietary supplements has risen exponentially. As throughout history, individuals are now turning to the use of "natural" therapies for the prevention, treatment and cure of almost every ailment and aging malady imaginable. often without substantial proof of safety or efficacy. One of the most popular herbal supplements is Ginkgo biloba extract, taken for its perceived "memory enhancing" properties. Given the inordinate popularity, growing use, and substantial number of pharmaceutical products containing G. biloba, coupled with demands for product safety and "hard evidence," science has followed this trend closely with an ever-expanding body of pharmacological and clinical data on such preparations. Claims that standardized G. biloba extract (EGb 761) can modulate the cellular environment of an organism under both physiological and stress conditions may be attributed to its multivalent or totipotent properties, and can now be substantiated by the availability of modern molecular techniques. As opposed to pharmacologically manufactured or synthetic drugs, which provide a single target for a single receptor as the mechanism of action, EGb 761 is able to up- or down-regulate signaling pathways, gene transcription, cellular metabolism, etc., and thus assist in the regulation of the general physiological status of the cell and/or organism in response to stressors posed by both intracellular and extracellular conditions. Presumably, this is one of the biggest advantages of using natural products for the prevention and treatment of infirmity, as well as the maintenance of health in an organism. PMID- 14740188 TI - Cerebrosides of Candida lipolytica yeast. AB - Candida lipolytica yeast was grown batchwise on glucose medium. Cerebrosides were isolated from the sphingolipid fraction of total lipids using column chromatography and separated into two compounds by high-performance thin-layer chromatography. Glucose was detected as the sole sugar constituent in cerebrosides. The fatty acid composition of cerebrosides was characterised by a predominance of saturated fatty acids and by a high proportion of fatty acids with 16 carbon atoms. The dominant fatty acid was h16:0. The principal long-chain base components of both cerebroside species were trihydroxy bases, 18- and 20 phytosphinosine. The unique characteristic of cerebrosides was the presence of a high proportion of sphingosine (one-fourth of the total long-chain bases), which is a common characteristic of mammalian sphingolipids and rarely occurs in yeast cerebrosides. The ceramide moiety profile of cerebrosides is similar to that of epidermal ceramides, which implies a possibility for their application in care cosmetics. PMID- 14740189 TI - Physico-chemical and transglucosylation properties of recombinant sucrose phosphorylase from Bifidobacterium adolescentis DSM20083. AB - Clones of a genomic library of Bifidobacterium adolescentis were grown in minimal medium with sucrose as sole carbon source. An enzymatic fructose dehydrogenase assay was used to identify sucrose-degrading enzymes. Plasmids were isolated from the positive colonies and sequence analysis revealed that two types of insert were present, which only differed with respect to their orientation in the plasmid. An open reading frame of 1,515 nucleotides with high homology for sucrose phosphorylases was detected on these inserts. The gene was designated SucP and encoded a protein of 56,189 Da. SucP was heterologously expressed in Escherichia coli, purified, and characterized. The molecular mass of SucP was 58 kDa, as estimated by SDS-PAGE, while 129 kDa was found with gel permeation, suggesting that the native enzyme was a dimer. The enzyme showed high activity towards sucrose and a lower extent towards alpha-glucose-1-phosphate. The transglucosylation properties were investigated using a broad range of monomeric sugars as acceptor substrate for the recombinant enzyme, while alpha-glucose-1 phosphate served as donor. D- and L-arabinose, D- and L-arabitol, and xylitol showed the highest production of transglucosylation products. The investigated disaccharides and trisaccharides were not suitable as acceptors. The structure of the transglucosylation product obtained with D-arabinose as acceptor was elucidated by NMR. The structure of the synthesized non-reducing dimer was alpha Glcp(1-->1)beta-Araf. PMID- 14740190 TI - Increased antifungal and chitinase specific activities of Trichoderma harzianum CECT 2413 by addition of a cellulose binding domain. AB - Trichoderma harzianum is a widely distributed soil fungus that antagonizes numerous fungal phytopathogens. The antagonism of T. harzianum usually correlates with the production of antifungal activities including the secretion of fungal cell walls that degrade enzymes such as chitinases. Chitinases Chit42 and Chit33 from T. harzianum CECT 2413, which lack a chitin-binding domain, are considered to play an important role in the biocontrol activity of this strain against plant pathogens. By adding a cellulose-binding domain (CBD) from cellobiohydrolase II of Trichoderma reesei to these enzymes, hybrid chitinases Chit33-CBD and Chit42 CBD with stronger chitin-binding capacity than the native chitinases have been engineered. Transformants that overexpressed the native chitinases displayed higher levels of chitinase specific activity and were more effective at inhibiting the growth of Rhizoctonia solani, Botrytis cinerea and Phytophthora citrophthora than the wild type. Transformants that overexpressed the chimeric chitinases possessed the highest specific chitinase and antifungal activities. The results confirm the importance of these endochitinases in the antagonistic activity of T. harzianum strains, and demonstrate the effectiveness of adding a CBD to increase hydrolytic activity towards insoluble substrates such as chitin rich fungal cell walls. PMID- 14740191 TI - Properties and applications of microbial transglutaminase. AB - Some properties and applications of the transglutaminase (TGase) referred to as microbial TGase (MTGase), derived from a variant of Streptomyces mobaraensis (formerly classified as Streptoverticillium mobaraense), are described. MTGase cross-linked most food proteins, such as caseins, soybean globulins, gluten, actin, myosins, and egg proteins, as efficiently as mammalian TGases by forming an epsilon-(gamma-glutamyl)lysine bond. However, unlike many other TGases, MTGase is calcium-independent and has a relatively low molecular weight. Both of these properties are of advantage in industrial applications; a number of studies have illustrated the potential of MTGase in food processing and other areas. The crystal structure of MTGase has been solved. It provides basic structural information on the MTGase and accounts well for its characteristics. Moreover, an efficient method for producing extracellular MTGase has been established using Corynebacterium glutamicum. MTGase may be expected to find many uses in both food and non-food applications. PMID- 14740192 TI - Degradation of polycyclic aromatic hydrocarbons by a newly isolated dibenzofuran utilizing Janibacter sp. strain YY-1. AB - The dibenzofuran (DF)-utilizing bacterium strain YY-1 was newly isolated from soil. The isolate was identified as Janibacter sp. with respect to its 16S rDNA sequence and fatty acid profiles, as well as various physiological characteristics. In addition to DF, strain YY-1 could grow on fluorene and dibenzothiophene as sole sources of carbon and energy. It was also able to cometabolize a variety of polycyclic aromatic hydrocarbons including dibenzo- p dioxin, phenanthrene, and anthracene. The major metabolites formed from DF, biphenyl, dibenzothiophene, and naphthalene were identified by using gas chromatography-mass spectrometry as 2,3,2'-trihydroxybiphenyl, biphenyl dihydrodiol, dibenzothiophene 5-oxide, and coumarin, respectively. These results indicate that strain YY-1 can catalyze angular dioxygenation, lateral dioxygenation, and sulfoxidation. PMID- 14740193 TI - Heterologous expression of metabotropic glutamate receptor subtype 1 in Saccharomyces cerevisiae. AB - The upstream region of the isocitrate lyase gene (UPR-ICL) from the n-alkane utilizing yeast Candida tropicalis serves as a useful promoter of gene expression in the yeast Saccharomyces cerevisiae. The production of rat metabotropic glutamate receptor 1alpha (mGluR1alpha), which belongs to the G-protein-coupled receptor (GPCR) family, was tested under the control of UPR-ICL. Expression of mGluR1alpha was found in recombinant clones and enhanced by replacing the signal sequence of mGluR1alpha with the corresponding region of the alpha-factor receptor (Ste2), which is a GPCR found in S. cerevisiae. Moreover, the membrane fraction from a recombinant clone associated with Vesl-1S/Homer-1a protein binds the mGluR1alpha in rat cerebellum. These results suggest that the UPR-ICL controlled gene expression system is useful for heterologous GPCRs in S. cerevisiae. PMID- 14740194 TI - Treatment of the yeast Rhodotorula glutinis with AlCl(3) leads to adaptive acquirement of heritable aluminum resistance. AB - When aluminum (Al) was added to a culture, growth of Rhodotorula glutinis IFO1125 was temporarily arrested, showing longer lag phases, depending on the Al concentrations (50-300 microM) added, but the growth rates were not affected at all. Resistant strains obtained by one round of plate treatment containing Al reverted the resistance level to the wild-type level when cultivated without Al. Repeated Al treatments, however, induced heritable and stable Al resistance, the level of which was increased up to 4,000 microM by stepwise increments in Al concentrations. Thus, the heritable Al resistance adaptively acquired was due neither to adaptation nor to mutation, but to a mechanism which has yet to be studied. Heritable Al resistance seemed to release the Al inhibition of magnesium uptake. PMID- 14740195 TI - Enhanced secretion of Bacillus stearothermophilus L1 lipase in Saccharomyces cerevisiae by translational fusion to cellulose-binding domain. AB - The secretion of Bacillus stearothermophilus L1 lipase in Saccharomyces cerevisiae was investigated by employing a fusion partner, a cellulose-binding domain (CBD) from Trichoderma harzianum endoglucanase II (THEG). The CBD was connected to the N-terminal of L1 lipase through an endogenous linker peptide from THEG. The expression cassette for the fusion protein in S. cerevisiae was constructed using the alpha-amylase signal peptide and the galactose-inducible GAL10 promoter. Secretion of CBD-linker-L1 lipase by this fusion construct was dramatically 7-fold enhanced, compared with that of the mature L1 lipase without CBD-fusion. The fusion protein was secreted into the culture medium, reaching levels of approximately 1.3 g/l in high-cell-density fed-batch cultures. Insertion of a KEX2 cleavage site into the junction between CBD-linker and L1 lipase resulted in the same level of enhanced secretion, indicating that the CBD linker fusion probably plays a critical role in secretion from endoplasmic reticulum to Golgi apparatus. Therefore, the CBD from THEG can be used both as an affinity tag and as a secretion enhancer for the secretory production of heterologous proteins in S. cerevisiae, since in vivo breakage at the linker was almost negligible. PMID- 14740196 TI - Improved xylanase production by Trichoderma reesei grown on L-arabinose and lactose or D-glucose mixtures. AB - Trichoderma reesei Rut C-30 was grown on eight different natural or rare aldopentoses as the main carbon source and on mixtures of an aldopentose with D glucose or lactose. The fungal cells consumed all aldopentoses tested, except L xylose and L-ribose. The highest total xylanase and cellulase activities were achieved when cells were grown on L-arabinose as the main carbon source. The total xylanase activity produced by cells grown on L-arabinose was even higher than that produced by cells grown on an equal amount of lactose. In co-metabolism of D-glucose (15 g l(-1)) and L-arabinose (5 g l(-1)), the total volumetric and specific xylanase productivities were improved (derepressed) approximately 23- and 18-fold, respectively, compared to a cultivation on only D-glucose (20 g l( 1)). In a similar experiment, in which cells were grown on a mixture of lactose and L-arabinose, the xylanase productivity was approximately doubled, compared to a cultivation on only lactose. The cellulase productivities, however, were not improved by the addition of L-arabinose. Compared with a typical industrial fungal enzyme production process with lactose as the main carbon source, better volumetric and specific xylanase productivities were achieved both on a lactose/arabinose mixture and on a glucose/arabinose mixture. PMID- 14740197 TI - Over-expression system for secretory phospholipase D by Streptomyces lividans. AB - The structural gene for phospholipase D (PLD) of an actinomycete, Streptoverticillium cinnamoneum, together with its promoter region was introduced into Streptomyces lividans using a shuttle vector-pUC702-for Escherichia coli and S. lividans. The transformant was found to secrete a large amount of PLD (about 2.0x10(4) U/l, 42 mg/l) when cultured in a jar fermentor. Both an initial glucose concentration of 17.5 g/l and the feeding of carbon and nitrogen sources are effective for efficient secretion of PLD; under these culture conditions, the amount of PLD secreted reached a maximum level (about 5.5x10(4) U/l, 118 mg/l) after about 60 h. In contrast to the original producer, Stv. cinnamoneum, which secretes only a small amount of PLD (about 1.1x10(3) U/l, 2 mg/l) along with other extracellular proteins, this heterologous expression system is markedly more efficient in production of secretory PLD. PMID- 14740199 TI - Oropharyngeal fetus in fetu. AB - Fetus in fetu is an extremely rare entity and refers to the growth of a "parasitic" twin within a more mature fetus due to its inclusion within cells of the blastocyst. The presence of a vertebral column is considered essential for the diagnosis of a fetus in fetu. The retroperitoneum is the most common location for a fetus in fetu and its location in the mouth is rare. We report a case of oropharyngeal mass detected on antenatal sonography with imaging features consistent with a fetus in fetu. Complete surgical removal of the mass was aided by imaging. Follow-up of these patients is recommended to detect the rare incidence of malignant degeneration. PMID- 14740198 TI - Structural and functional studies of the nicotinic acetylcholine receptor by solid-state NMR. AB - Over the last seven years, solid-state NMR has been widely employed to study structural and functional aspects of the nicotinic acetylcholine receptor. These studies have provided detailed structural information relating to both the ligand binding site and the transmembrane domain of the receptor. Studies of the ligand binding domain have elucidated the nature and the orientation of the pharmacophores responsible for the binding of the agonist acetylcholine within the agonist binding site. Analyses of small transmembrane fragments derived from the nicotinic acetylcholine receptor have also revealed the secondary structure and the orientation of these transmembrane domains. These experiments have expanded our understanding of the channel's structural properties and are providing an insight into how they might be modulated by the surrounding lipid environment. In this article we review the advances in solid-state NMR applied to the nicotinic acetylcholine receptor and compare the results with recent electron diffraction and X-ray crystallographic studies. PMID- 14740200 TI - Age-related normal ranges for the Haller index in children. AB - PURPOSE: The Haller index is an accepted CT method for evaluating thoracic dimensions in patients with pectus excavatum. The purpose of this study is to establish age- and gender-related norms for the Haller index in childhood. MATERIALS AND METHODS: We retrospectively reviewed 574 consecutive chest CT examinations (M=285, F=289) performed at our institution from August 2001 through March 2002. Seventeen patients with a history of chest-wall deformity, trauma, or syndrome were excluded, for a total sample size of 557 patients. The Haller index was calculated for each patient, using electronic calipers. The sample population was then separated by gender and placed into 2-year age groupings. Two-way analysis of variance and Tukey's multiple comparisons were performed to determine significance at a=0.05. The least-square mean Haller index values for each age group and gender were calculated with 95% confidence intervals. RESULTS: In both males and females, the 0- to 2-year age group showed a significantly smaller mean Haller index than older children. In addition, females had significantly greater Haller index values than males in the 0- to 6- and 12- to 18-year age groups. CONCLUSION: The Haller index, a quantitative measurement of chest-wall configuration, demonstrates significant age- and gender-related variability. This should be considered when evaluating the patient with suspected chest-wall deformity. PMID- 14740201 TI - Extra-osseous involvement of Langerhans' cell histiocytosis in children. AB - The predominant clinical and radiological features of Langerhans' cell histiocytosis (LCH) in children are due to osseous involvement. Extra-osseous disease is far less common, occurring in association with bone disease or in isolation; nearly all anatomical sites may be affected and in very various combinations. The following article is based on a multicentre review of 31 children with extra-osseous LCH. The objective is to summarise the diverse possibilities of organ involvement. The radiological manifestations using different imaging modalities are rarely pathognomonic on their own. Nevertheless, familiarity with the imaging findings, especially in children with systemic disease, may be essential for early diagnosis. PMID- 14740202 TI - Is the frontal radiograph alone sufficient to evaluate for pneumonia in children? AB - BACKGROUND: In our cost- and radiation-conscious environment, the feasibility of performing only a frontal radiograph for the diagnosis of pneumonia in children needs to be reassessed. OBJECTIVE: To determine the diagnostic efficacy of the frontal radiograph alone in comparison to the frontal and lateral combined radiographs for the radiographic diagnosis of pneumonia in children. MATERIALS AND METHODS: Three radiologists retrospectively and independently reviewed the frontal radiographs alone and separately reviewed the frontal and lateral radiographs of 1,268 children referred from the emergency room for chest radiographs. A majority interpretation of at least two radiologists for the frontal views alone was compared with majority interpretation of the frontal and lateral combined views for the radiographic diagnosis of pneumonia. "Pneumonia" was defined as a focus of streaky or confluent lung opacity. RESULTS: For the radiographic diagnosis of pneumonia, the sensitivity and specificity of the frontal view alone were 85% and 98%, respectively. For the confluent lobar type of pneumonia, the sensitivity and specificity increased to 100%. CONCLUSION: When the frontal view alone yields a diagnosis of confluent lobar pneumonia, this is highly reliable. However, nonlobar types of infiltrates will be underdiagnosed in 15% of patients using the frontal view alone. The clinical impact of these radiographically underdiagnosed pneumonias needs to be assessed prior to implementing the practice of using only frontal radiographs for diagnosing pneumonia. PMID- 14740203 TI - In vivo amyloid imaging in Alzheimer's disease. AB - Targeted approaches to therapy for Alzheimer's disease have evolved based on detailed understanding of the genetic, molecular biologic, and neuropathologic basis of the disease. Given the potential for greater treatment efficacy in the earlier stages of the disease, the notion of early diagnosis has become more relevant. Current clinical and imaging diagnostic approaches lack reliability in the preclinical and prodromal phases of the disease. We review emerging studies on imaging of the molecular substrate of the disease, most notably the amyloid peptide, which hope to increase early diagnostic efficacy. We offer a brief overview of the demographics, diagnostic criteria, and current imaging tests, followed by a review of amyloid biology and developments in cerebral amyloid imaging yielded by recent in vitro, in vivo and human studies. PMID- 14740204 TI - Amplitude changes of unconditioned eyeblink responses in patients with cerebellar lesions. AB - Timing and amplitude parameters of unconditioned eyeblink responses were investigated in 24 patients with unilateral cerebellar lesions following infarcts within the territory of the superior cerebellar artery (SCA, n=12) and of the posterior inferior cerebellar artery (PICA, n=12). The extent of cortical cerebellar lesions, i.e., which lobules were affected and possible involvement of cerebellar nuclei, was determined by three-dimensional magnetic resonance imaging (3D MRI). Amplitude parameters of eyeblink responses were normalized and expressed as percentage of the unaffected side in patients and the second tested side in age-matched controls. Normalized peak amplitudes, burst area and burst duration were significantly increased in SCA patients with lesions restricted to cortical areas. Burst onset and time to peak were not significantly different compared with controls. Temporal and amplitude parameters of eyeblink responses were unchanged in SCA patients with additional involvement of cerebellar nuclei and in patients with lesions of the PICA territory. Consistent with animal lesion and recording studies and a recent human functional magnetic resonance imaging (fMRI) study, the present data suggest that cortical areas of the superior cerebellum are of importance in eyeblink control in humans. These areas partly overlap with areas known to be critical in eyeblink conditioning. PMID- 14740205 TI - Asymmetrically acting lycopene beta-cyclases (CrtLm) from non-photosynthetic bacteria. AB - Carotenoids have important functions in photosynthesis, nutrition, and protection against oxidative damage. Some natural carotenoids are asymmetrical molecules that are difficult to produce chemically. Biological production of carotenoids using specific enzymes is a potential alternative to extraction from natural sources. Here we report the isolation of lycopene beta-cyclases that selectively cyclize only one end of lycopene or neurosporene. The crtLm genes encoding the asymmetrically acting lycopene beta-cyclases were isolated from non photosynthetic bacteria that produced monocyclic carotenoids. Co-expression of these crtLm genes with the crtEIB genes from Pantoea stewartii (responsible for lycopene synthesis) resulted in the production of monocyclic gamma-carotene in Escherichia coli. The asymmetric cyclization activity of CrtLm could be inhibited by the lycopene beta-cyclase inhibitor 2-(4-chlorophenylthio)-triethylamine (CPTA). Phylogenetic analysis suggested that bacterial CrtL-type lycopene beta cyclases might represent an evolutionary link between the common bacterial CrtY type of lycopene beta-cyclases and plant lycopene beta- and epsilon-cyclases. These lycopene beta-cyclases may be used for efficient production of high-value asymmetrically cyclized carotenoids. PMID- 14740206 TI - Immunity in rainbow trout, Oncorhynchus mykiss, against the monogenean Discocotyle sagittata following primary infection. AB - Rainbow trout (Oncorhynchus mykiss) were experimentally infected by continuous or single exposure with the monogenean Discocotyle sagittata. To determine whether immunity follows primary infection, fish were exposed to a secondary challenge by one of two modes: (1) primary infections were cleared with praziquantel (PZQ) and hosts re-infected with 100 oncomiracidia; (2) parasites were allowed to reach maturity and hosts super-infected with 100 oncomiracidia. Fish challenged after initial continuous exposure developed significant partial resistance to re infection, carrying burdens 35% lower than controls. PZQ treatment controls demonstrated that the drug did not account for the protection observed. Single exposure did not significantly modify the outcome of secondary challenges. Super infection experiments suggested that no concomitant immunity develops. No correlation was found between initial burdens and the outcome of secondary challenges in the same individual. Significantly elevated anti-D. sagittata antibodies were detected in infected fish, but there was no correlation between immunoglobulin levels and parasite burdens. PMID- 14740207 TI - Chemoembolization of rat liver metastasis with irinotecan and quantification of tumor cell reduction. AB - PURPOSE: Hematogenic metastasis of patients with colorectal cancer most frequently effects the liver; the prognosis of affected patients is dramatically worsened by the presence of this lesion. The aim of this study was to evaluate the effect of hepatic arterial chemoembolization (HACE) with irinotecan versus 5 fluorouracil as a standard agent in a rat liver metastasis model. MATERIALS AND METHODS: Diffuse liver metastasis was induced by injecting 4 x 10(6) CC531-lac-Z rat colorectal carcinoma cells into the portal vein of male Wag/Rij rats. Irinotecan (10 mg/kg, 30 mg/kg, and 60 mg/kg) and 5-fluorouracil (40 mg/kg, 60 mg/kg, and 90 mg/kg) were administered concomitantly with degradable starch microspheres (30 mg/kg) for temporary embolization. The tumor cell load was determined quantitatively using a chemoluminescence assay. RESULTS: HACE with irinotecan induced a complete remission in 44% of the animals and the highest dose reduced the mean tumor cell load by 66% (P < 0.001). In contrast, the highest dose of 5-FU caused a reduction of only 18% (P = 0.026) and altogether 23% complete remissions were observed in response to 5-FU. The sensitivity of CC531-lac-Z cells versus irinotecan (IC50 32 pM after 72 h) and 5-FU (IC50 80 microM) mirrored the effects observed in vivo on a qualitative basis. CONCLUSION: In conclusion, the effect of HACE with irinotecan surpassed that of HACE with 5 FU and prompts further investigation in clinical trials. PMID- 14740209 TI - The evolutionary origin of animal cellulose synthase. AB - Urochordates are the only animals that produce cellulose, a polysaccharide existing primarily in the extracellular matrices of plant, algal, and bacterial cells. Here we report a Ciona intestinalis homolog of cellulose synthase, which is the core catalytic subunit of multi-enzyme complexes where cellulose biosynthesis occurs. The Ciona cellulose synthase gene, Ci-CesA, is a fusion of a cellulose synthase domain and a cellulase (cellulose-hydrolyzing enzyme) domain. Both the domains have no animal homologs in public databases. Exploiting this fusion of atypical genes, we provided evidence of a likely lateral transfer of a bacterial cellulose synthase gene into the urochordate lineage. According to fossil records, this likely lateral acquisition of the cellulose synthase gene may have occurred in the last common ancestor of extant urochordates more than 530 million years ago. Whole-mount in situ hybridization analysis revealed the expression of Ci-CesA in C. intestinalis embryos, and the expression pattern of Ci-CesA was spatiotemporally consistent with observed cellulose synthesis in vivo. We propose here that urochordates may use a laterally acquired "homologous" gene for an analogous process of cellulose synthesis. PMID- 14740210 TI - Cloning and developmental expression of MARK/Par-1/MELK-related protein kinase xMAK-V in Xenopus laevis. AB - MAK-V/Hunk is a recently identified MARK/Par-1-related mammalian protein kinase. Although the precise function of this protein kinase is yet to be established, available data suggest its involvement in animals' development and in the physiology of the nervous system. Here we report characterization of a cDNA encoding Xenopus laevis orthologue of MAK-V/Hunk protein kinase, xMAK-V. The in silico analysis also revealed MAK-V/Hunk orthologues in the fish Fugu rubripes and primitive chordate Ciona intestinalis but not in invertebrate species such as Drosophila melanogaster and Caenorhabditis elegans, suggesting that MAK-V/Hunk is a chordate-specific protein kinase. The expression of xmak-v in X. laevis embryos was analyzed using whole-mount in situ hybridization. Expression of xmak-v has been detected in all developmental stages studied including maternal expression in unfertilized eggs. The xmak-v mRNA has a predominant occurrence on the animal hemisphere of the egg, and this pattern of expression is sustained throughout cleavage and blastula stages. At the gastrula stage xmak-v expression is restricted to the ectoderm. In the later stage embryos xmak-v is expressed over the entire embryonic surface including the open neural plate at stage 15 and also in neural tube at stage 22. At tadpole stage xmak-v expression is strong in embryonic epidermis, nervous system and sensory organs, and is also obvious in perisomitic mesoderm and brachial arches. PMID- 14740211 TI - Biosynthesis of methionine-derived glucosinolates in Arabidopsis thaliana: recombinant expression and characterization of methylthioalkylmalate synthase, the condensing enzyme of the chain-elongation cycle. AB - The major class of glucosinolates in Arabidopsis thaliana (L.) Heynh. are biosynthesized from methionine involving a three-step chain-elongation cycle. Each passage through the cycle results in the net addition of a single methylene group, with up to six cycles of elongation occurring in A. thaliana. The first reaction of the cycle is catalyzed by a methylthioalkylmalate synthase (MAMS), which condenses a omega-methylthio-2-oxoalkanoic acid with acetyl-CoA. Here we have demonstrated that MAM1, one of two similar genes in the A. thaliana ecotype Columbia, encodes a MAMS catalyzing the condensing reactions of the first two elongation cycles but not those of further cycles. The Columbia ecotype is dominated by compounds that have undergone only two elongation cycles. The A. thaliana MAM1 protein exhibits basic sequence similarity to other previously described enzymes catalyzing the condensation of 2-oxo acids and acetyl-CoA, such as isopropylmalate synthase (EC 2.3.3.13), an enzyme of leucine biosynthesis, and homocitrate synthase (EC 2.3.3.14). It also shares similar properties with them, including the catalytic requirements for a divalent metal ion and an adenine nucleotide. However, the MAM1 protein does not show activity with the substrates of any of these other enzymes, and was chromatographically separable from isopropylmalate synthase in extracts of A. thaliana. Thus, MAM1 is exclusively an enzyme of secondary metabolism, distinct from primary metabolic enzymes catalyzing similar reactions. PMID- 14740212 TI - Molecular cloning and expression analysis of three genes encoding starch synthase II in rice. AB - Three starch synthase (SS) genes, OsSSII-1, OsSSII-2 and OsSSII-3, were identified in rice (Oryza sativa L.) and localized to chromosomes 10, 2 and 6, respectively. The three OsSSII full-length cDNAs were cloned, and the predicted amino acid sequences were found to share 52-73% similarity with other members of the plant SSII family. The SS activity of each OsSSII was confirmed by expression and enzyme activity assay in Escherichia coli. Expression profile analysis revealed that OsSSII-1 was expressed in endosperms, leaves and roots; OsSSII-2 was mainly expressed in leaves, while OsSSII-3 was mainly expressed in endosperms. Similar to the OsSSI proteins, the OsSSII-2 and OsSSII-3 proteins were found in the soluble as well as the starch-granule-bound fractions in rice. The roles of the OsSSII proteins in starch biosynthesis in rice and the evolutionary relationships of the genes encoding monocotyledonous and dicotyledonous class-II SS enzymes are discussed. PMID- 14740213 TI - Cloning and functional expression of an ( E, E)-alpha-farnesene synthase cDNA from peel tissue of apple fruit. AB - Increased production of terpenes and many other aroma-related volatiles occurs with the onset of ripening in apple ( Malus domestica Borkh.) fruit. The gaseous plant hormone ethylene plays a key role in the induction of volatile synthesis, but the mechanism is not yet understood. Using a degenerate primer based on a short conserved sequence shared by several sesquiterpene synthases, reverse transcription-polymerase chain reaction with RNA isolated from peel tissue of 'Law Rome' apples yielded an approx. 800-bp gene fragment. This was used to screen a cDNA library generated from the peel tissue mRNA. A full-length terpene synthase (TS) cDNA 1,931 nucleotides long was isolated. The 1,728-bp open reading frame encodes a protein 576 amino acids long with a molecular mass of 66 kDa. Sequence analysis of the apple TS showed it to be most similar to several monoterpene synthases. Oddly, the TS includes an RR(X(8))W motif near the N terminus that is common among monoterpene synthases but it lacks the plastid transit peptide sequence typically associated with genes of that group. Expression of the apple TS gene in Escherichia coli gave myc-epitope-tagged and untagged proteins estimated at approx. 68 and approx. 66 kDa, respectively. In assays of sesquiterpene synthase activity, with farnesyl diphosphate as substrate, the untagged bacterially expressed TS gene product synthesized ( E, E) alpha-farnesene almost exclusively. In monoterpene synthase assays, with geranyl diphosphate as substrate, the untagged apple TS produced only ( E)-beta-ocimene, albeit at much reduced levels. Addition of a C-terminal myc tag appeared to completely prevent production of soluble protein under all of the expression conditions tested. This is the first report of an ( E, E)-alpha-farnesene synthase gene ( AFS1; GenBank accession number AY182241) from a flowering plant. RNA gel blots showed that AFS1 transcript increased about 4-fold in peel tissue of apple fruit during the first 4 weeks of storage at 0.5 degrees C. In contrast, when fruit were treated at harvest with 1-methylcyclopropene, a blocker of ethylene action, AFS1 mRNA declined sharply over the initial 4 weeks of cold storage, and fell to nearly undetectable levels by 8 weeks. PMID- 14740214 TI - NADH-dependent metabolism of nitric oxide in alfalfa root cultures expressing barley hemoglobin. AB - Transgenic alfalfa ( Medicago sativa L.) root cultures expressing sense and antisense barley ( Hordeum vulgare L.) hemoglobin were examined for their ability to metabolize NO. Extracts from lines overexpressing hemoglobin had approximately twice the NO conversion rate of either control or antisense lines under normoxic conditions. Only the control line showed a significant increase in the rate of NO degradation when placed under anaerobic conditions. The decline in NO was dependent on the presence of reduced pyridine nucleotide, with the NADH-dependent rate being about 2.5 times faster than the NADPH-dependent rate. Most of the activity was found in the cytosolic fraction of the extracts, while only small amounts were found in the cell wall, mitochondria, and 105,000- g membrane fraction. The NADH-dependent NO conversion exhibited a broad pH optimum in the range 7-8 and a strong affinity to NADH and NADPH ( K(m) 3 microM for both). It was sensitive to diphenylene iodonium, an inhibitor of flavoproteins. The activity was strongly reduced by applying antibodies raised against recombinant barley hemoglobin. Extracts of Escherichia coli overexpressing barley hemoglobin showed a 4-fold higher rate of NO metabolism as compared to non-transformed cells. The NADH/NAD and NADPH/NADP ratios were higher in lines underexpressing hemoglobin, indicating that the presence of hemoglobin has an effect on these ratios. They were increased under hypoxia and antimycin A treatment. Alfalfa root extracts exhibited methemoglobin reductase activity, using either cytochrome c or recombinant barley hemoglobin as substrates. There was a correspondence between NO degradation and nitrate formation. The activity was eluted from a Superose 12 column as a single peak with molecular weight of 35+/-4 kDa, which corresponds to the size of the hemoglobin dimer. The results are consistent with an NO dioxygenase-like activity, with hemoglobin acting in concert with a flavoprotein, to metabolize NO to nitrate utilizing NADH as the electron donor. PMID- 14740215 TI - Microtubules and microfilaments coordinate to direct a fountain streaming pattern in elongating conifer pollen tube tips. AB - This study investigates how microtubules and microfilaments control organelle motility within the tips of conifer pollen tubes. Organelles in the 30-microm long clear zone at the tip of Picea abies (L.) Karst. (Pinaceae) pollen tubes move in a fountain pattern. Within the center of the tube, organelles move into the tip along clearly defined paths, move randomly at the apex, and then move away from the tip beneath the plasma membrane. This pattern coincides with microtubule and microfilament organization and is the opposite of the reverse fountain seen in angiosperm pollen tubes. Application of latrunculin B, which disrupts microfilaments, completely stops growth and reduces organelle motility to Brownian motion. The clear zone at the tip remains intact but fills with thin tubules of endoplasmic reticulum. Applications of amiprophosmethyl, propyzamide or oryzalin, which all disrupt microtubules, stop growth, alter organelle motility within the tip, and alter the organization of actin microfilaments. Amiprophosmethyl inhibits organelle streaming and collapses the clear zone of vesicles at the extreme tip together with the disruption of microfilaments leading into the tip, leaving the plasma membrane intact. Propyzamide and oryzalin cause the accumulation of membrane tubules or vacuoles in the tip that reverse direction and stream in a reverse fountain. The microtubule disruption caused by propyzamide and oryzalin also reorganizes microfilaments from a fibrillar network into pronounced bundles in the tip cytoplasm. We conclude that microtubules control the positioning of organelles into and within the tip and influence the direction of streaming by mediating microfilament organization. PMID- 14740216 TI - Hairy root induction of Papaver somniferum var. album, a difficult-to-transform plant, by A rhizogenes LBA 9402. AB - Two strains of Agrobacterium rhizogenes (15834, LBA 9402) and one Agrobacterium tumefaciens strain [GV 3101 (PMP90RK, p35SGUS-2)] and four culture media were tested and compared for their ability to induce hairy root formation on wounded Papaver somniferum L. hypocotyls. Five weeks after the infection with A. rhizogenes LBA 9402, hairy roots appeared on 80% of the hypocotyls maintained in the hormone-free liquid medium. Six hairy-root cultures were established. Transformation was confirmed by polymerase chain reaction analysis. One clone was analysed for its alkaloid production. The total alkaloid content was higher in the transformed roots (0.46+/-0.06% DW) than in the untransformed roots (0.32+/ 0.05% DW). The transformed roots accumulated three times more codeine (0.18+/ 0.02% DW) than intact roots (0.05+/-0% DW). Moreover, morphine (0.255+/-0.03% DW) and sanguinarine (0.014+/-0% DW) were found in the liquid culture medium. PMID- 14740217 TI - Training effect on performance, substrate balance and blood lactate concentration at maximal lactate steady state in master endurance-runners. AB - Training effects on time-to-exhaustion, substrate and blood lactate balances at the maximal lactate steady state velocity (MLSSv) were examined. Eleven male, veteran, long-distance runners performed three tests before and after 6 weeks of training at MLSSv: an incremental test to determine maximum O2 uptake (VO(2,max)) and the velocity at the lactate threshold (vLT), a sub-maximal test of two stages of 20 min at 95 and 105% of vLT separated by 40 min rest to determine the MLSSv and the corresponding lactate concentration (MLSSc) and a time-to-exhaustion run at MLSSv for which the substrate balance was calculated. Duration and distance run at MLSSv increased dramatically respectively from 44+/-10 to 63+/-12 min and from 10.4 to 15.7 km respectively (P<0.01). MLSSv increased significantly with training but the relative fraction of VO(2,max) remained the same (85.2+/-4.5 vs. 85.3+/-5.2%, P=0.93). MLSSc was unaffected by training as determined from the percentage of energy yielded by carbohydrates (80%) during the exhaustive run at MLSSv. These findings show that training at MLSS elicits small increases in MLSSv and VO(2,max), but enhances time-to-exhaustion (endurance) at MLSSv substantially (+50%). Training does not change the proportion of carbohydrate oxidized, which is the major substrate used during an exhaustive run at MLSS lasting 1 h. PMID- 14740218 TI - Electrophysiological characterization and functional importance of calcium activated chloride channel in rat uterine myocytes. AB - In order to better understand the mechanisms underlying excitation of the uterus, we have elucidated the characteristics and functional importance of Ca(2+) activated Cl(-) currents ( I(Cl-Ca)) in pregnant rat myometrium. In 101/320 freshly isolated myocytes, there was a slowly inactivating tail current (162+/-48 pA) upon repolarization following depolarising steps. This current has a reversal potential close to that for chloride, and was shifted when [Cl(-)] was altered. It was activated by Ca(2+) (but not Ba(2+)) entry through L-type Ca(2+) channels, enhanced by the Ca(2+) channel agonist Bay K8644 (2 microM), and inhibited by the Cl(-) channel blockers, niflumic acid (10 microM) and anthracene-9-carboxylic acid (9-AC, 100 microM). We therefore conclude that the pregnant rat myometrium contains Ca(2+)-activated Cl(-) channels producing inward current in ~30% of its cells. When these channels were inhibited by niflumic acid or 9-AC in intact tissues, the frequency of spontaneous contractions, was significantly reduced. Niflumic acid was also shown to inhibit oxytocin-induced contractions and Ca(2+) transients. Neither 9-AC nor niflumic acid had any effect on high-K-invoked contractions. Taken together these data suggest that Ca(2+)-activated Cl(-) channels are activated by Ca(2+) entry and play a functionally important role in myometrium, probably by contributing to membrane potential and firing frequency (pacemakers) in these cells. PMID- 14740219 TI - European surgical community: the unification of surgical sciences anticipated by German-Polish surgeons in the 19th century. PMID- 14740220 TI - Cork workers' occupational asthma: lack of association with allergic sensitisation to fungi of the work environment. AB - OBJECTIVES: To evaluate allergic sensitisation to Chrysonilia sitophila, Penicillium glabrum, and Trichoderma longibrachiatum in cork workers with asthma. METHODS: Skin prick tests with a battery of common allergens and with the three fungi were performed on ten cork workers with asthma and eight non-exposed asthmatics. Based on serial peak expiratory flow measurements, five were classified as having occupational asthma (AO) and five as having non-occupational asthma (NOA). In exposed patients, specific antibodies for the three fungi were also studied by immunoblotting RESULTS: Two out of ten patients with occupational exposure and four out of eight of the control group showed positive results for skin prick tests for common allergens. Moreover, two out of five patients with OA and three out of eight controls exhibited sensitisation to storage mites. All exposed patients (with OA or NOA) had negative skin prick test results for the fungal extracts. In patients with asthma and occupational exposure, immunoblotting results confirmed the absence of specific IgE. However, specific IgG4 was present in some cases. CONCLUSIONS: Atopy does not seem to characterise occupational asthma in cork workers. Despite their long exposure to moulds, we could not find evidence of IgE sensitisation to the three most prevalent cork fungi in patients with OA, which points to the search for other causative agents, such as cork chemical compounds or contaminants. PMID- 14740221 TI - Trans-placental exposure of neonates to acrylamide--a pilot study. AB - OBJECTIVES: Acrylamide (AA) (CAS No 79-06-1) has most recently been identified as a food-borne toxicant generated during the heating process of starch-containing foods. It was the aim of the present study to investigate the trans-placental exposure of newborn infants to this possible human carcinogen by analysis of the specific haemoglobin adduct of AA ( N-2-carbamoylethylvaline, AAV) in the blood of mothers and the corresponding umbilical cord blood of neonates as a parameter of biochemical effects. METHODS: We investigated the blood of 11 women advanced in pregnancy (one smoker, ten non-smokers) and the corresponding umbilical cord blood of neonates for the N-terminal haemoglobin adducts of AA (AAV) and the smoking-specific adduct of acrylonitrile (CAS No 107-13-1) ( N-cyanoethylvaline, CEV). The limit of detection (LOD) was 5 pmol/g globin for AAV and 4 pmol/g globin for CEV. RESULTS: AAV could be determined in all blood samples of the mothers (median 21 pmol/g globin, range 18-104 pmol/g globin) as well as in the umbilical cord blood of neonates (median 10 pmol/g globin, range 6-43 pmol/g globin). The highest values were detected in the blood of the smoking mother and her child. CEV was detected only in the blood of the smoking mother (185 pmol/g globin) and the corresponding umbilical cord blood (69 pmol/g globin). DISCUSSION: AAV adduct levels in non-smoking mothers and neonates showed a good correlation (r=0.859). The concentration of AA adducts in the blood of neonates is approximately 50% of the adduct level found in the blood of the mother. In view of the shorter life span of neonatal erythrocytes and the lower body weight of newborn infants, the relative internal dose of AA in neonates (in microgrammes per kilogramme body weight) must be assumed to be at least equal to that of the mother. Because of the high cell-replication rates during foetal development, trans-placental exposure of neonates to AA might raise concerns. Neonates of smoking mothers take up much higher doses of AA than those of non-smoking mothers. PMID- 14740222 TI - Subcellular immunolocalisation of fatty acid translocase (FAT)/CD36 in human type 1 and type-2 skeletal muscle fibres. AB - Limited information exists about the putative role and expression in human skeletal muscle cells of the 88-kDa integral membrane protein fatty acid translocase (FAT), highly homologous to the human leucocyte differentiation factor CD36. Therefore, we investigated in healthy male individuals the muscle (m. vastus lateralis) fibre type specific expression and subcellular localisation of FAT/CD36. For this purpose four different monoclonal antibodies raised against human and mouse FAT/CD36 were used. Acetone or methanol/acetone fixation were tested. Serial cryosections were cut at -20 degrees C, thaw-mounted on uncoated glass slides and air-dried before processing indirect immunofluorescence assays. Images were examined in a Nikon ER800 microscope, digitally captured, processed and analysed by LUCIA laboratory software. Three antibodies showed that FAT/CD36 was: (1) most abundantly expressed in capillary endothelium, (2) colocalised with caveolin-3, which indicates that FAT/CD36 is in the sarcolemma, or its close vicinity, and (3) abundantly expressed in (or in the close vicinity) of the sarcolemma and intracellular structures of type-1 muscle fibres, and much less abundantly in the sarcolemma of type-2 muscle fibres. One of the antibodies raised against mouse CD36 also detected myosin heavy chain 1, which makes it unsuitable in skeletal muscle research. The fixation (acetone or methanol/acetone) was found to be highly important for the result. PMID- 14740223 TI - Evidence for a membrane carbonic anhydrase IV anchored by its C-terminal peptide in normal human pancreatic ductal cells. AB - The high concentration of HCO(3)(-) ions (150 mM) in the human pancreatic ducts raises the question of the membrane proteins responsible for their secretion in addition to the Cl(-)/HCO(3)(-) exchanger. In this study, we investigated the expression of carbonic anhydrase IV (CA IV), a possible candidate. Experiments were carried out on specimens of normal human pancreas obtained from brain-dead donors ( n=9) as well as on isolated human ductal cells. Two antibodies were generated: CA IV NH(2) antibody directed against the NH(2) terminal of human glycosyl phosphatidylinositol (GPI)-anchored CA IV and CA IV COOH antibody directed against the COOH terminal of the same protein before its association with a GPI in the rough endoplasmic reticulum. A 35-kDa CA IV was detected in the homogenates of human pancreas. Immunocytochemistry demonstrated the expression of CA IV in centroacinar cells and in intercalated, intralobular, and interlobular ductal cells. The immunoreactivity observed with the CA IV COOH antibody was mainly localized on luminal membranes of ductal cells. Treatment of purified plasma membranes with phosphatidylinositol-phospholipase C indicated that the CA IV expressed in pancreatic ducts was not GPI-anchored. Its detection in the same extracts by the CA IV COOH antibody indicated that it was anchored by a hydrophobic segment at the carboxy terminal. Taken together, these results suggest that normal human pancreatic ductal cells express a 35-kDa CA IV anchored in their luminal plasma membrane by a hydrophobic segment of the COOH terminus. In view of its localization and its mode of anchorage in luminal plasma membranes, this CA IV may participate in the maintenance of luminal pH. PMID- 14740224 TI - Expression of intermediate filaments of podocytes within nephrotic syndrome glomerulopathies in children. AB - The study attempted to define characteristics of renal podocytes in nephrotic syndrome glomerulopathies in children with and without glomerular immaturity based on the histochemical expression of cytokeratin 18 (CK 18) and vimentin. Material consisted of 29 renal biopsies performed in the Department of Pediatric Nephrology, Poznan University of Medical Sciences, between 1991 and 2000. The study group included 16 children with mesangial glomerulonephritis (MesGN) and signs of glomerular immaturity and 13 children with MesGN without signs of glomerular immaturity. The control tissue was derived from macroscopically normal renal cortex taken from kidneys resected for localised neoplasms ( n=3). In the control samples, the immunocytochemical expression of CK 18 was found only in epithelial cells of proximal and distal tubules. Vimentin was present in all podocytes, some mesangial cells and endothelium. In all cases of children with MesGN with signs of immaturity, both CK 18-positive and vimentin-positive podocytes were found. In all cases of MesGN without immaturity we revealed CK 18 negative podocytes but with distinct vimentin-positive expression. Reorganisation of cytoskeletal proteins within immature podocytes may be associated with the unfavourable clinical course of nephrotic syndrome in children. The application of antibodies against intermediate filaments may help to differentiate between mature and immature forms of MesGN. PMID- 14740225 TI - Association between lens opacities and mortality in the Priverno Eye Study. AB - BACKGROUND: Lens opacities are associated with a higher risk of death, although there are some discrepancies regarding the specific types of cataract representing risk. The purpose of the present study was to further investigate the relationships between different types of lens opacity and patient survival. METHODS: In 1987, 860 residents of Priverno, Italy, aged 45-69 years underwent an ophthalmologic examination. Based on patient histories and the findings of the slit-lamp examination, each of the 860 patients was classified according to the type of opacity (pure cortical, pure nuclear, pure posterior subcapsular, mixed, and surgical aphakia). The survivors of the original cohort were re-examined in 1994. Death and survival rates were computed by the Kaplan-Meier method. Associations between mortality and significant factors were included in a stepwise Cox proportional-hazards regression model. RESULTS: Forty-four members of the original cohort had died during the 7-year follow-up. Age-adjusted survival curves based on Kaplan-Meier estimates showed significantly lower survival in those whose baseline examinations had revealed pure nuclear opacity (log rank test: P=0.020) and aphakia (log rank test: P<0.001). When adjusted for other mortality risk factors (age, sex, diabetes, cardiovascular diseases), the hazard ratio was 4.32 for pure nuclear opacity (95% CI 1.13-16.4) and 18.3 for aphakia (95% CI 3.21-104.0). CONCLUSIONS: The analysis of the Priverno data seems to confirm an association between lower survival and cataracts, particularly those confined to the lens nucleus and those that had already prompted surgery. PMID- 14740226 TI - Comparison of the growth potential of retinal pigment epithelial cells obtained during vitrectomy in patients with age-related macular degeneration or complex retinal detachment. PMID- 14740227 TI - Population genetics of the 15 AmpF lSTR Identifiler loci in Kosovo Albanians. AB - The 15 AmpF lSTR Identifiler loci (D8S1179, D21S11, D7S820, CSF1PO, D3S1358, THO1, D13S317, D16S539, D2S1338, D19S433, VWA, TPOX, D18S51, D5S818 and FGA) were analyzed in a sample of 136 unrelated Albanian adults from Kosovo. The agreement with HWE was confirmed for all loci with the exception of TPOX (based on the exact test only). The combined power of discrimination (PD) and the combined power of exclusion (PE) for the 15 studied loci were 0.99999999999999997 and 0.9999995, respectively. According to the presented data, FGA proved to be the most informative marker. An interpopulation comparison between Kosovo Albanians and Croatians (as an example of a population from the Balkans) revealed significant differences in four out of nine loci. PMID- 14740229 TI - Influence of spinal hypotension on fetal oxidative status during elective cesarean section in uncomplicated pregnancies. AB - METHODS: We examined the relation between spinal hypotension (systolic blood pressure: <100 mmHg or <80% of the baseline value) and fetoplacental oxygen free radicals during elective cesarean section. Plasma xanthine, serum uric acid and plasma malondialdehyde levels in umbilical venous blood and blood gases in the umbilical artery were measured in patients receiving spinal anesthesia for elective cesarean section complicated with (n=26) and without (n=26) spinal hypotension. Patients with spinal hypotension were divided to two groups on the duration of the hypotension: those with the duration of <2 min (n=19) and those with the duration of > or =2 min (n=7). RESULTS: There were no measurable differences in these variables between the control and the hypotension with the duration of <2 min groups. While, the plasma xanthine, serum uric acid and plasma malondialdehyde levels in the hypotension group with the duration of > or =2 min were significantly higher than those in the control group without the significant differences in blood gases and pH-levels. PMID- 14740228 TI - Nuclear bodies and compartmentalization of pre-mRNA splicing factors in higher plants. AB - We studied the fine structural organization of nuclear bodies in the root meristem during germination of maize and Arabidopsis thaliana using electron microscopy (EM). Cajal bodies (CBs) were observed in quiescent embryos and germinating cells in both species. The number and distribution of CBs were investigated. To characterize the nuclear splicing domains, immunofluorescence labelling with antibodies against splicing factors (U2B" and m3G-snRNAs) and in situ hybridisation (with U1/U6 antisense probes) were performed combined with confocal microscopy. Antibodies specific to the Arabidopsis SR splicing factor atRSp31 were produced. AtRSp31 was detected in quiescent nuclei and in germinating cells. This study revealed an unexpected speckled nuclear organization of atRSp31 in root epidermal cells where micro-clusters of interchromatin granules were also observed by EM. Therefore, we examined the distribution of green fluorescent protein (GFP)-tagged atRSp31 in living cells after Agrobacterium -mediated transient expression. When expressed transiently, atRSp31-GFP exhibited a speckled distribution in leaf cells. Treatments with alpha-amanitin, okadaic acid, staurosporine or heat shock induced the speckles to reorganize. Furthermore, we generated stable Arabidopsis transgenics expressing atRSp31-GFP. The distribution of the fusion protein was identical to that of endogenous atRSp31. Three-dimensional time-lapse confocal microscopy showed that speckles were highly dynamic domains over time. PMID- 14740230 TI - Expression of cyclin D1 in normal, hyperplastic and neoplastic endometrium and its correlation with Ki-67 and clinicopathological variables. AB - METHODS: We investigated cyclin D1 expression in proliferative endometrium, endometrial hyperplasia and endometrioid adenocarcinoma, and examined the correlation of cyclin D1 expression with Ki67 as a cell proliferation associated marker. Immunohistochemical expression of cyclin D1 and Ki67 were studied in 30 cases with endometrial carcinoma, 14 cases with atypical hyperplasia, 15 cases with simple hyperplasia and 30 cases with proliferative endometrium. RESULTS: One out of 30 patients (3.3%) with proliferative endometrium, 1 out of 14 patients (7.1%) with atypical hyperplasia, and 8 out of 30 patients (26.6%) with endometrial carcinoma were found to have immunoreactivity to cyclin D1. All cases of simple hyperplasia had negative staining for cyclin D1. A positive immunoreaction for Ki67 was obtained in all cases. Statistically significant difference was found in cyclin D1 immunoreactivity between both proliferative endometrium and adenocarcinoma, and simple hyperplasia and adenocarcinoma (p<0.05). In patients with adenocarcinoma, cyclin D1 immunoreactive cases had higher mean Ki67 values compared with the non-immunoreactive ones (p<0.05). Ki67 and cyclin D1 immunoreactivity had no impact on overall survival. Univariate analysis revealed a significant relationship between survival and grade and stage (p<0.01). Cyclin D1 expression was not correlated with age, depth of myometrial invasion, lymphovascular space involvement, grade, lymph node metastasis and stage. CONCLUSION: Cyclin D1 expression in endometrial carcinoma is higher than proliferative endometrium and simple hyperplasia. These findings support that cyclin D1 may play a role in endometrial carcinogenesis. PMID- 14740231 TI - Glutathione S-transferase polymorphisms in patients with drug eruption. AB - Glutathione S-transferase (GST) enzymes play an important role in drug metabolism. GST is a multigene family of enzymes involved in the detoxification and in a few instances activation of a wide variety of chemicals. Detoxification features make it plausible to search for GST polymorphism in patients with drug eruption. The GSTM (mu), GSTT (theta) and GSTP (pi) have been shown to be polymorphically distributed. The GSTT1, GSTM1 and GSTP1 gene polymorphism were detected using real-time PCR. GSTM1 and GSTT1 null genotypes were found to be associated with an increased risk of drug eruption (OR 2.27, 95% CI 1.20-5.21; OR 2.48, 95% CI 1.12-6.39, respectively). No relationship was observed between the null combination of the GSTM1 and GSTT1 genotype polymorphisms and drug eruption risk (OR 2.65, 95% CI 0.62-11.25). Our results show that GSTP1 polymorphism is not a significant contributor to drug eruption risk. The GSTM1 and GSTT1 gene polymorphisms seem to be associated with the development of drug eruption. Further studies may shed additional light on the role of GSTM1, GSTT1 and GSTP1 in drug eruption. PMID- 14740232 TI - Total hip arthroplasty after avascular necrosis of the femoral head: does etiology affect the results? AB - INTRODUCTION: The aim of the study was to evaluate the results of total hip arthroplasty (THA) due to avascular necrosis (AVN) of the femoral head and to establish whether the cause of AVN affects the results. MATERIAL AND METHODS: A group of 68 patients, 17-82 years of age (mean: 49.9 years) underwent 84 total hip arthroplasties due to AVN. The patients were divided into subgroups according to the etiology of AVN of the hip joint. The results of each group were evaluated by the Harris Hip score (HHS) at 3-18 years (mean: 6.2 years). The complication rate was also assessed. RESULTS: The mean preoperative HHS was 28.5+/-4.5 as opposed to a postoperative HHS of 86+/-10. The revision rate was 16.7%. Etiology does not affect the final outcome, but less favorable long-term results were found in the steroid-induced AVN patients. CONCLUSIONS: Despite the previously reported, less favorable results, THA is a good option for the younger population, even with AVN, especially in bilateral disease. Etiology did not affect the final outcome. However, patients with steroid-induced AVN should be informed that although their final functional results will equal those of other groups, the longevity of the implants is limited. PMID- 14740235 TI - Lipid management with statins. The lower the better? AB - The primary and not yet accomplished goal remains to treat all patients with coronary heart disease to the LDL cholesterol target < 100 mg/dl. To date there is no conclusive evidence for a recommendation of a LDL cholesterol goal lower than 100 mg/dl for all patients. Patients with high vascular risk benefit from statin therapy irrespective of cholesterol levels, underlining the importance of the assessment of global vascular risk as the basis of modern lipid therapy. PMID- 14740236 TI - Atheromatous disease of the thoracic aorta and systemic embolism. Clinical picture and therapeutic challenge. AB - Systemic embolism is a frequent cause of stroke. At the beginning of the last decade by introduction of transesophageal echocardiography and other imaging techniques atheromatosis of the aortic arch has been recognized as an important source of embolism. Formerly in the pre-TEE era, this entity was included into cryptogenic strokes. Aortic atheromas are found in about one quarter of patients presenting with embolic events. The severity of atherosclerosis graded by TEE correlates with the risk for future embolism, especially if mobile lesions or superimposed thrombi are present. Independent of plaque extension, patients with unstable plaques characterized by echo-lucency, inhomogenity, lacking of calcifications, ulceration, mobile parts and concomitant spontaneous echo contrast within the aorta have a higher risk for embolic events. However, the diagnosis of aortic atheromatosis is mostly established if an embolic event has already occurred. Therefore, it is important to identify patients at risk, especially before they undergo interventions with manipulation at the aorta like coronary bypass surgery. Risk factors are age above 70, diabetes mellitus, hyperlipidemia, arterial hypertension, aortic calcifications on standard chest X ray, elevated serum levels of C-reactive protein, other inflammatory markers, and an activated coagulation. Randomized studies for treatment of patients with severe aortic atheromatosis are not yet existing. Warfarin has been shown to prevent stroke in patients with mobile atheromas and superimposed thrombi, but there are case reports about aggravation of cholesterol embolism under warfarin treatment. It is concluded from other atherosclerotic manifestations that plaque stabilizing treatment with statins and ACE inhibitors is also beneficial. PMID- 14740237 TI - Perspectives and new approaches for improving cardiopulmonary resuscitation in adults beyond current guidelines. AB - Setting clear priorities for the sequence and importance of actions during cardiopulmonary resuscitation (CPR) is of utmost importance for future guidelines. Unless performed under the rare condition of hypoxic arrest, combined compression and ventilation is usually not necessary in one-rescuer resuscitation of adults. After notifying the emergency medical services (EMS), precordial compression at a rate of 100/min is just as effective or may even be preferable in the majority of cases caused by arrhythmic arrest. Considering the pathophysiological and experimental evidence, chest compression has proven to be more important, even in multi-rescuer settings, than resuscitation ventilation with its problems and risks. International recommendations for compression without respiration for rescuers unwilling to perform resuscitation ventilation or for so-called telephone CPR were not included in the guidelines of the European Resuscitation Council (ERC) probably for reasons of brevity and simplification. However, training for basic cardiopulmonary resuscitation of adults with cardiac arrest should also stress the importance of chest compression over ventilation. Moreover, current studies controversially discuss the optimal time point of defibrillation after collapse. These findings point to the enormous demand for research in the field of cardiopulmonary resuscitation. PMID- 14740238 TI - Catheter-based therapy for hypertrophic obstructive cardiomyopathy. First in hospital outcome analysis of the German TASH Registry. AB - BACKGROUND: Registry results of the new catheter-based method in the treatment for HOCM are missing so far. In 1997, the Transcoronary Ablation of Septal Hypertrophy Registry (TASH Registry) was established by the German Cardiac Society (GCS) as a multicenter, national registry of patients with HOCM undergoing the new catheter interventional therapy. This is the report of the in hospital outcome of patients who underwent the procedure during the first two years of data collection in the registry. METHODS AND RESULTS: Information was based on three standard forms for each patient, with a total of 86 variables. Information was collected on an "intention to treat" basis. The TASH Registry includes the establishment of a data base in the data collecting center. Ten centers participated. Enrollment forms were received for 264 patients out of 279 patients registered up to January 2000. There was a history of medical treatment of 3.6+/-3.9 years. The vast majority of patients (91%) were treated in three centers. The Vasalva maneuver and the exercise Doppler echocardiography were used for noninvasive stress testing. Exercise Doppler echocardiography induced a significantly higher augmentation of the baseline gradient (70.1% vs 133.4%; p<0.01). The echo-contrast guided technique was used for the intervention in 50.8% and the pressure angiography guided technique in 49.2%. On the average 2.8+/-1.3 ml of alcohol were injected. Before the procedure, the gradient measured by catheterization was 60.4+/-38.6 mmHg at baseline and 142.7+/-46.2 mmHg following the extrasystolic beat. At the end of the session it was reduced significantly by 75% and 67%. The peak phosphocreatine kinase activity was 482.5+/-246.4 U/L. Major complications occurred in 15.6% including a mortality rate of 1.2% and a permanent pacemaker implantation rate because of total heart block in 9.6%. There was an early in-hospital improvement of dyspnoe corresponding to a significant decrease of NYHA functional class from 2.8+/-0.7 to 1.8+/-0.6 (p<0.001). Similar hemodynamic and clinical benefit was found in patients with and without resting gradient at baseline. CONCLUSION: This analysis for the first time gives a comprehensive overview of clinical characteristics, technique, procedural data, in-hospital outcome and complications in a large number of patients with HOCM who were treated by the new catheter-based method and prospectively enrolled in a registry. The results contribute considerably to critical evaluation and validation of the new technique. This analysis supports the catheter-based method to constitute a new therapeutic option for very symptomatic patients, to be effective both in patients with and without intraventricular pressure gradient at rest and to be an alternative to surgical treatment, as has been stated recently. PMID- 14740239 TI - Prognostic significance of the signal averaged electrocardiogram in patients with chronic stable coronary artery disease. Analysis in the time domain and by spectral temporal mapping. AB - BACKGROUND: Ventricular late potentials detected by the signal averaged electrocardiogram (SAECG) have been used to predict cardiac death in patients after recent myocardial infarction. The goal of this study was to investigate the prognostic significance of the SAECG in a population of chronic coronary artery disease, with and without previous myocardial infarction. METHODS: SAECG was recorded in 698 patients with angiographically proven coronary artery disease and analyzed by time domain analysis (TDA) and by spectral temporal mapping (STM). Cardiac death or ventricular fibrillation (= cardiac event) were used as the primary endpoint for follow-up (25 to 33 months). RESULTS: A cardiac event occurred in 46 out of 698 patients (6.6%). An abnormal SAECG using TDA was found in 43% of patients with a cardiac event, as compared to 21.7% in those without (p<0.0005). The probability of a cardiac event during follow-up was 4.4% when TDA and STM were both normal, 9.5% and 10.2% when either STM or TDA were abnormal and 28.5% when both were abnormal. A duration of the averaged QRS complex of more than 120 ms and a left ventricular ejection fraction of less than 45% were the only independent predictors of a cardiac event. Logistic regression analysis could predict a cardiac event with a sensitivity of 54% and a specificity of 88%. CONCLUSIONS: In patients with chronic coronary artery disease the duration of the signal averaged QRS complex and left ventricular ejection fraction are independent predictors of cardiac death or ventricular fibrillation. PMID- 14740240 TI - Does statin therapy influence steroid hormone synthesis? AB - Statins reduce cholesterol and isoprenoid de novo biosynthesis as well as receptor mediated uptake of cholesterol for steroidogenesis. The present randomized placebo-controlled trial investigated whether pravastatin (40 mg/day) reduces the plasma concentrations of steroid hormones as well as of gonadotropins. Patients (n = 22; 15 males, 7 females) were treated with pravastatin (40 mg/day) or placebo. Levels of total and LDL cholesterol, the steroid hormones estradiol, testosterone, cortisol and dehydroepiandrosterone sulphate (DHEAS) as well as FSH and LH were studied. Pravastatin led to a significant reduction of total cholesterol and LDL cholesterol. There was no significant change in estradiol, testosterone, cortisol or DHEAS plasma concentrations. There was no compensatory change in FSH or LH. It is concluded that pravastatin does not alter steroid hormones or gonadotropins in a clinically applicable dose, which significantly reduces total and LDL cholesterol. PMID- 14740241 TI - [Simultaneous carotid endarterectomy and cardiac surgery-additional risk factor or safe procedure?]. AB - BACKGROUND: The occurrence of severe carotid artery disease in more than 12% of patients requiring ACB results in a discrepancy concerning the best treatment for both diseases. We reviewed the early outcome of patients with ACB and/or valve replacement and simultaneous carotid endarterectomy (TEA). METHODS: We evaluated retrospectively 244 patients operated simultaneously between 7/94 and 10/2001: 209 patients received ACB, 35 patients ACB and/or valve replacement. Mean age was 68 years. 188 patients were male. We analyzed risk factors, morbidity, incidence of neurological complications and 30 day mortality. RESULTS: Perioperative stroke with hemiplegia occurred in 3.3% (8 patients). Of these patients, 4 showed contralateral carotid artery occlusion, 2 contralateral severe stenosis. Two patients (0.8%) experienced prolonged reversible ischemic neurological deficit (PRIND), 4 patients (1.6%) transient ischemic attack (TIA). 30-day morbidity was 4.5%. Three patients died due to low cardiac output, 6 patients due to extracardial reasons, 2 patients (0.8) due to cerebral death. CONCLUSION: Simultaneous TEA and cardiac surgery can be performed with an acceptable risk for neurological complications and mortality. Occlusion of the contralateral carotid artery could be identified as an evident predictor for increased neurological complications. Compared to two-stage procedures, combined operations yield a reduction of hospital costs. PMID- 14740242 TI - [Bland-White-Garland syndrome in an adult]. AB - We report about a 47-year-old woman, who presented with a history of cardiac failure. Echocardiography showed an impaired left ventricular function, clinically significant mitral regurgitation and pulmonary hypertension. Diagnosis of a Bland- White-Garland syndrome was made by coronary angiography. Subsequent therapy consisted of ligation of the anomalus origin of the left coronary artery, implantation of a Mammaria interna graft to the left coronary artery and replacement of the mitral valve by a mechanical prosthesis. One year after operation, left ventricular function was still impaired. At a 3-year follow-up, left ventricular function improved continuously. PMID- 14740243 TI - [Persistent angina pectoris in spite of successful myocardial revascularisation]. AB - A 73-year-old obese woman underwent coronary artery-bypass operation in 11/1995 because of a coronary two vessel disease. The left coronary artery was bypassed by the left mammarial internal artery. In 2 and 3/2002, balloon-dilatation of stenoses of the right coronary artery and the circumflex was performed. Angina pectoris relapsed and in 9/2002 the patient was admitted to our hospital with tentative diagnosis of restenosis. Physical investigation showed a blood pressure of the right arm of 160/80 and of the left arm of 120/ 80 mmHg. Coronarography showed the three vessel disease known since 2/2002 with a restenosis of the right coronary artery which was immediately treated by balloon-dilatation and stent implantation. Colour duplex-sonography of the carotid and subclavian arteries revealed extraordinary plaques and a reduced flow of the left vertebral artery. The left subclavian artery could only be seen distal to the discharge of the vertebral artery and showed a poststenotic flow. The patient had angina pectoris when carrying out personal hygiene already 2 days after balloon-dilatation and stent-implantation. ECG showed new aspects. Coronarography showed no relapse of stenosis, but 70% stenosis of the left subclavian artery with a marked coronary steal-syndrome. In 10/ 2002, the patient underwent balloon-dilatation and stent implantation of the subclavian stenosis and became free of complaints. Coronary steal-syndrome can be the reason for persistent angina pectoris in spite of successful coronary artery-bypass operation with a mammarial internal bypass. It is absolutely necessary to take blood pressure from both arms to recognise a possible stenosis of the subclavian artery which can be the key to all. PMID- 14740244 TI - Neovascularization in left atrial myxoma. AB - Myxoma is the most common benign tumor of the heart. Tumor neovascularization at the angiographically detectable level is thought to be rare. We report on a 78 year-old female patient with a left atrial myxoma showing marked tumor vascularity. This observation had consequences to the therapeutic management: during tumor excision, the large supplying artery had to be ligated. PMID- 14740245 TI - [Guidelines: acute coronary syndrome (ACS). 1: ACS without persistent ST segment elevations]. PMID- 14740248 TI - Intralesional bleomycin injection (IBI) treatment for haemangiomas and congenital vascular malformations. AB - Successful treatment of vascular anomalies has eluded the physician until now, despite various treatments utilised. Bleomycin has been successfully used in intralesional injection treatment of cystic hygromas and haemangiomas, based specifically on a high sclerosing effect on vascular endothelium. In a prospective study of 95 patients, the effectiveness of intralesional bleomycin injection (IBI) treatment in haemangiomas and vascular malformations was evaluated and documented. Complete resolution or significant improvement occurred in 80% of all patients treated. Complete resolution occurred in 49% of haemangiomas, 32% of venous malformations, and 80% of cystic hygromas. Significant improvement occurred in 38% of haemangiomas, 52% of venous malformations, 13% of cystic hygromas and 50% of lymphatic malformations. Of the six patients who presented with a painful lesion, four experienced complete resolution and two had significant improvement to treatment. Local complications encountered were superficial ulceration occurring in 2 patients, and cellulitis in 1 of the 95 patients. Systemic complications were flu-like symptoms in three patients and partial, transient hair loss in two patients. None of the patients presented with haematological toxic effects or signs of pulmonary involvement (fibrosis, hypertension). IBI is an effective treatment in haemangiomas and vascular malformation lesions, obviating the need for invasive primary surgery or systemic treatment regimens in 80% of cases, and allowing for limited need of secondary surgical or adjunctive procedures in cases with a moderate result. PMID- 14740249 TI - Horner's syndrome resulting from agenesis of the internal carotid artery: report of a third case. AB - INTRODUCTION: Only two previous reports of agenesis of the internal carotid artery with a coexistent Horner's syndrome are to be found in the medical literature. CASE REPORT: We report a case of a male child seen initially for what was presumed to be a traumatic superficial temporal artery aneurysm. MRA incidentally revealed ipsilateral agenesis of the internal carotid artery. This child's physical exam included ipsilateral Horner's syndrome. CONCLUSION: This case report illustrates that agenesis of the internal carotid artery does indeed cause Horner's syndrome in some cases. The clinician who diagnoses a patient with Horner's syndrome should remember that agenesis of the internal carotid artery may be the cause as this entity can be associated in itself with other pathologies of the central nervous system. PMID- 14740250 TI - Kinetic analyses of waterborne Ca and Cd transport and their interactions in the gills of rainbow trout (Oncorhynchus mykiss) and yellow perch (Perca flavescens), two species differing greatly in acute waterborne Cd sensitivity. AB - We evaluated the differential nature of interactions between waterborne Ca and Cd transport in the gills of yellow perch (Perca flavescens) and rainbow trout (Oncorhynchus mykiss), two species with a more than 400-fold difference in acute waterborne Cd tolerance. The Jmax (maximum rate of uptake) and K(m) (inverse of affinity) for Ca uptake, in the absence of Cd, were significantly lower in yellow perch (120.48 nM g(-1) wet wt h(-1) and 92.17 microM, respectively) relative to rainbow trout (188.68 nM g(-1) wet wt h(-1) and 243.90 microM, respectively). Similarly, the Jmax for Cd uptake, at the lowest waterborne Ca level (100 microM) tested, was significantly lower in yellow perch (0.27 nM g(-1) wet wt h(-1)) relative to rainbow trout (0.40 nM g(-1) wet wt h(-1), but no significant difference was observed in the K(m) values between the two species (yellow perch: 32.47 nM; rainbow trout: 31.27 nM). Waterborne Cd (0-890 nM) as well as waterborne Ca (100-1,000 microM) competitively inhibited branchial uptake of each other in both species. However, analyses of inhibitor constants for branchial Ca uptake by waterborne Cd (Ki[Cd2+]) revealed that the inhibition was about 1.8 times more potent in rainbow trout compared to yellow perch. In contrast, analyses of inhibitor constants for branchial Cd uptake by waterborne Ca (Ki[Cd2+]) indicated that the inhibition was more than three fold more potent in yellow perch than in rainbow trout. Higher branchial Ca uptake and more potent inhibition by Cd as well as higher branchial Cd uptake and less potent inhibition by Ca were also reflected in whole-body measurements of Ca and Cd influx in trout relative to perch. Overall, whole-body effects were in accord with the branchial kinetic analyses. These results further strengthen the conclusion that branchial influxes of Ca and Cd occur through common pathways. Moreover, interspecific differences in acute waterborne Cd sensitivity can be explained, at least in part, by the differential nature of interactions between waterborne Ca and Cd transport in fish gills. PMID- 14740251 TI - Hierarchy and social status in Budongo chimpanzees. AB - The status hierarchy is fundamental in the lives of male chimpanzees. This study describes the dominance interactions and social status among adult male chimpanzees of the Sonso community in the Budongo Forest Reserve, Uganda, during the period that they were first studied (1994 and 1995). Social dominance is typically measured using the behaviour of either the subordinate or the dominant individual, but a relationship is dependent on the behaviour of both parties and this study explicitly used both subordinate and dominant behaviours to investigate the status hierarchy. Among adult males of the Sonso community, agonistic interactions occurred at a low rate and pant-grunts were rare, but males could be ranked into separate hierarchies of agonistic dominance and pant grunting (labelled 'respect') using ratios of behaviour performed/behaviour received. These hierarchies were combined to form a single hierarchy of social status that divided the males among five distinct status levels. The highest status level was held by an alliance between two males who replaced the previous alpha male during the first part of the study. Neither male in this alliance partnership pant-grunted to the other, although the reason for cooperative behaviour was unclear. Although the nominally beta male was treated as such by other adult males, he achieved surprisingly little mating success. Budongo Forest chimpanzees do not warrant the sometimes-expressed view that they are non aggressive and peaceable and the broad pattern of their status interactions matches with that seen in other chimpanzee populations. PMID- 14740252 TI - Lovastatin inhibits its own synthesis in Aspergillus terreus. AB - Lovastatin suppresses its own synthesis in the microfungus Aspergillus terreus. The inhibitory effect was documented by spiking identical batch cultures with pure lovastatin (0, 50, 100 and 250 mg/l) 24 h after initiation from spores. PMID- 14740253 TI - Hsp70 and Hsp90--a relay team for protein folding. AB - Molecular chaperones are a functionally defined set of proteins which assist the structure formation of proteins in vivo. Without certain protective mechanisms, such as binding nascent polypeptide chains by molecular chaperones, cellular protein concentrations would lead to misfolding and aggregation. In the mammalian system, the molecular chaperones Hsp70 and Hsp90 are involved in the folding and maturation of key regulatory proteins, like steroid hormone receptors, transcription factors, and kinases, some of which are involved in cancer progression. Hsp70 and Hsp90 form a multichaperone complex, in which both are connected by a third protein called Hop. The connection of and the interplay between the two chaperone machineries is of crucial importance for cell viability. This review provides a detailed view of the Hsp70 and Hsp90 machineries, their cofactors and their mode of regulation. It summarizes the current knowledge in the field, including the ATP-dependent regulation of the Hsp70/Hsp90 multichaperone cycle and elucidates the complex interplay and their synergistic interaction. PMID- 14740255 TI - Comparative DNA sequence analysis of mapped wheat ESTs reveals the complexity of genome relationships between rice and wheat. AB - The use of DNA sequence-based comparative genomics for evolutionary studies and for transferring information from model species to related large-genome species has revolutionized molecular genetics and breeding strategies for improving those crops. Comparative sequence analysis methods can be used to cross-reference genes between species maps, enhance the resolution of comparative maps, study patterns of gene evolution, identify conserved regions of the genomes, and facilitate interspecies gene cloning. In this study, 5,780 Triticeae ESTs that have been physically mapped using wheat ( Triticum aestivum L.) deletion lines and segregating populations were compared using NCBI BLASTN to the first draft of the public rice ( Oryza sativa L.) genome sequence data from 3,280 ordered BAC/PAC clones. A rice genome view of the homoeologous wheat genome locations based on sequence analysis shows general similarity to the previously published comparative maps based on Southern analysis of RFLP. For most rice chromosomes there is a preponderance of wheat genes from one or two wheat chromosomes. The physical locations of non-conserved regions were not consistent across rice chromosomes. Some wheat ESTs with multiple wheat genome locations are associated with the non-conserved regions of similarity between rice and wheat. The inverse view, showing the relationship between the wheat deletion map and rice genomic sequence, revealed the breakdown of gene content and order at the resolution conferred by the physical chromosome deletions in the wheat genome. An average of 35% of the putative single copy genes that were mapped to the most conserved bins matched rice chromosomes other than the one that was most similar. This suggests that there has been an abundance of rearrangements, insertions, deletions, and duplications eroding the wheat-rice genome relationship that may complicate the use of rice as a model for cross-species transfer of information in non-conserved regions. PMID- 14740256 TI - 3' rapid amplification of cDNA ends (RACE) walking for rapid structural analysis of large transcripts. AB - The 3' rapid amplification of cDNA ends (3' RACE) is widely used to isolate the cDNA of unknown 3' flanking sequences. However, the conventional 3' RACE often fails to amplify cDNA from a large transcript if there is a long distance between the 5' gene-specific primer and poly(A) stretch, since the conventional 3' RACE utilizes 3' oligo-dT-containing primer complementary to the poly(A) tail of mRNA at the first strand cDNA synthesis. To overcome this problem, we have developed an improved 3' RACE method suitable for the isolation of cDNA derived from very large transcripts. By using the oligonucleotide-containing random 9mer together with the GC-rich sequence for the suppression PCR technology at the first strand of cDNA synthesis, we have been able to amplify the cDNA from a very large transcript, such as the microtubule-actin crosslinking factor 1 (MACF1) gene, which codes a transcript of 20 kb in size. When there is no splicing variant, our highly specific amplification allows us to perform the direct sequencing of 3' RACE products without requiring cloning in bacterial hosts. Thus, this stepwise 3' RACE walking will help rapid characterization of the 3' structure of a gene, even when it encodes a very large transcript. PMID- 14740259 TI - Incidence of residual intracranial AVMs after surgical resection and efficacy of immediate surgical re-exploration. AB - BACKGROUND: The true incidence of residual lesions after surgical resection of AVMs is not well documented in the literature. Partial surgical resection is thought to not confer any improvement over the natural history risk of hemorrhage of AVMs, and in certain cases may actually increase the risk of hemorrhage. Over the past 11 years, we have adopted a policy of immediate postoperative angiography with immediate surgical re-exploration if a residual lesion is seen. The purpose of the present study was to review our experience to determine the incidence of residual lesions and subsequent outcome. METHODS: From June 1991 to June 2002, 324 patients underwent craniotomy and surgical AVM resection. As per protocol, all patients underwent immediate postoperative angiography. We have a protocol for immediate surgical re-exploration if a residual lesion is seen on postoperative angiographic exam. FINDINGS: There were total six patients (1.8% of patients operated with intracranial AVMs) with residual lesions on postoperative angiography. All six patients underwent immediate surgical re-exploration with complete 100% obliteration; two patients required two re-exploration procedures. There was one operative complication: posterior cerebral artery and superior cerebellar artery infarcts after re-exploration of residual lesion after surgical resection of a large occipito-temperal-parietal AVM. There were no other morbidities and no mortalities. CONCLUSIONS: The incidence of residual lesions seen on postoperative angiography after AVM surgery at an experienced center is 1.8%. Because of the potential imminent danger of hemorrhage from a residual lesion, we recommend a policy of immediate postoperative angiography (or intraoperative angiography if image quality is satisfactory) for all AVM surgery and early surgical re-exploration if a residual lesion is seen. PMID- 14740260 TI - Surgical resection of gliomas WHO grade II and III located in the opercular region. AB - BACKGROUND: Surgery in the opercular region especially in the dominant hemisphere impose a major challenge for the neurosurgeon due to the close vicinity to functional important motor and speech areas. The purpose of the present study is to analyse on a homogenous patient group pre- and postoperative functional deficits with regard to different speech qualities (e.g. aphasia, apraxia), and to correlate these data with MR and intraoperative monitoring results. METHOD: Fourteen patients with suspected low grade astrocytomas in the opercular region consecutively treated by surgery were eligible for this study (histology revealed 3 WHO grade III tumours). Degree and duration of postoperative deficits were retrospectively evaluated according to tumour location and boundaries on MR, intraoperative neuromonitoring results and extent of tumour resection. FINDINGS: Postoperatively, 8 patients showed speech or language disturbances, in 4 patients combined with motor deficits mainly of the contralateral upper extremity. Fifty percent of the neuropsychologically tested patients exhibited speech apraxia while the other 50% had a true aphasic syndrome. Recovery of the latter deficits was in general faster and more complete. The severity and duration of postoperative deficits was in good correlation with the distance of the resection margin to the next positive stimulation point(s), and a distance of more than 0.5 cm proved to avoid major impairments. The distribution of functional important stimulation points in relation to the tumour extension was not predictable, and - unexpectedly -- up to 50% of these sites were found overlaying the tumour. INTERPRETATION: Surgery for WHO grade II and III gliomas in the opercular region can result in speech apraxia or an aphasic syndrome with or without concomitant motor deficits. Intraoperative cortical electrical stimulation is essential in resecting tumours in the opercular region to avoid permanent morbidity. PMID- 14740254 TI - Arabidopsis kinome: after the casting. AB - Arabidopsis thaliana is used as a favourite experimental organism for many aspects of plant biology. We capitalized on the recently available Arabidopsis genome sequence and predicted proteome, to draw up a genome-scale protein serine/threonine kinase (PSTK) inventory. The PSTKs represent about 4% of the A. thaliana proteome. In this study, we provide a description of the content and diversity of the non-receptor PSTKs. These kinases have crucial functions in sensing, mediating and coordinating cellular responses to an extensive range of stimuli. A total of 369 predicted non receptor PSTKs were detailed: the Raf superfamily, the CMGC, CaMK, AGC and STE families, as well as a few small clades and orphan sequences. An extensive relationship analysis of these kinases allows us to classify the proteins in superfamilies, families, sub-families and groups. The classification provides a better knowledge of the characteristics shared by the different clades. We focused on the MAP kinase module elements, with particular attention to their docking sites for protein-protein interaction and their biological function. The large number of A. thaliana genes encoding kinases might have been achieved through successive rounds of gene and genome duplications. The evolution towards an increasing gene number suggests that functional redundancy plays an important role in plant genetic robustness. PMID- 14740261 TI - Distinct supratentorial lesions mimicking cerebral gliomas. AB - BACKGROUND: Various supratentorial pathological conditions can mimic neoplastic cerebral lesions clinically as well as radiologically. Analysis of the neuroradiological findings, the clinical history, laboratory and other paraclinical data mostly help to narrow down the diagnosis of cerebral pathologies. Sometimes, however, histopathological analysis of the operative specimen after surgery reveals unexpected findings. PATIENTS AND FINDINGS: In a series of 197 operative procedures performed for glioma surgery between August 2000 and August 2002 we found six distinct cases mimicking supratentorial tumours. Clinicoradiological findings had suggested a neoplastic gliomatous process in all cases. But histopathological examination revealed that in reality one patient had been affected by a stroke, two by hypertensive encephalopathy, and one by radiation necrosis; multiple sclerosis was the underlying pathology in two patients. INTERPRETATION: Conditions which show features similar to those of neoplastic cerebral lesions require advanced magnetic resonance imaging (MRI). The benefit of such sophisticated imaging in relation to the costs is an important issue in this context. Further research in the field of modern image modalities is necessary to evaluate these noninvasive techniques for specification of intracerebral lesions. PMID- 14740262 TI - Impact of primary surgery on pituitary function in patients with non-functioning pituitary adenomas -- a study on 721 patients. AB - INTRODUCTION: The aim of this study was to define the impact of surgery on pituitary function in a large consecutive series of patients harbouring non functioning pituitary adenomas. MATERIALS AND METHOD: Between December 1982 and December 2000, a total of 822 patients underwent primary surgery in the authors' department. In 721 cases a complete set of endocrinological data was available. Functions of the pituitary-gonadal, pituitary-thyroid and pituitary-adrenal axes were assessed immediately before surgery and again one week, 3 months and 1 year after the operation, utilizing standardized tests and commercially available assays. RESULTS: There was some degree of pre-operative hypopituitarism in 561 (85%) and 53 (86.3%) of the patients belonging to the transsphenoidal and the transcranial groups, respectively. Prior to transsphenoidal [transcranial] surgery, 163 (31%) [34 (55.7%)] of the patients had secondary adrenal deficiency, 463 (76.6%) [49 (89%)] had hypogonadism and 105 (19.1%) [14 (25.4%)] were hypothyroid. Preoperatively, prolactin levels were mildly elevated in 167 patients (25.3%), whereas 1 year after surgery, levels were elevated in only 5 patients. Permanent diabetes insipidus occurred in 4 patients, 2 from the transsphenoidal group (0.3%) and 2 from the transcranial group (3.2%). Following transsphenoidal surgery 110 (19.6%) of patients had normal pituitary function [versus 0% after transcranial surgery], 169 (30.1%) [6 (11.3%)] showed improvement, 274 (48.9%) [49 (73.7%)] had persistent deficits and 8 (1.4%) [8 (15%)] showed deterioration of pituitary function. DISCUSSION: These data indicate that transsphenoidal surgery for non-functioning pituitary adenomas in expert hands is, relatively, far less detrimental to patients compared with transcranial surgery. The latter carries a much greater risk of post-operative deterioration in pituitary function. PMID- 14740263 TI - The Ki-67 proliferation antigen in meningiomas. Experience in 600 cases. AB - BACKGROUND: Meningiomas are mostly benign tumours that can be cured by surgical resection. Because meningiomas tend to recur, long term management in patients with subtotal tumour resection remains controversial. Previous studies have shown that the proliferation potential of meningiomas by Ki-67 labelling indices (LI) might predict their natural history. The purpose of this study was to analyse the reliability of Ki-67-labelling index in predicting the behaviour of meningiomas, and to help the neurosurgeon in establishing better follow up criteria and long term management strategies for these patients. METHOD: From 1990 to 2000 1328 meningiomas have been operated in our Neurosurgical Department. A total of 600 tumours were examined immunohistochemically using the Mib-1 monoclonal antibody. Clinical charts of the patients including surgical, histological and follow up records, as well as imaging studies were analysed retrospectively. Ki-67 LI were correlated with neuroradiological findings, 3D volumetric studies, histological subtype, recurrence-free survival, grade of resection, consistency of tumour tissue, location, osseous involvement, en plaque appearance, vascularity and progesterone-receptor status. FINDINGS: Among the 600 patients analysed, there were 66% females (mean LI 3.8%) and 34% males (mean LI 5.7%), including 20 neurofibromatosis-type-2 (NF-II) patients with a mean LI of 5.2%. Histological grading revealed 91% WHO degrees I meningiomas (mean LI 3.28%), 7% WHO degrees II (mean LI 9.95%) and 2% WHO degrees III (mean LI 12.18%). Labelling indices in recurrent meningiomas increased from initial resection to a fourth local resection. A significant correlation between negative progesteron-receptor status and high tumour vascularity with high Ki-67 LI was seen. Ki67 was not a statistically significant predictor of survival time in totally excised WHO degrees I meningiomas. INTERPRETATION: Mib-1 is one important tool in addition to routine histological evaluation, but a combination of clinical factors and particularly the extent of surgical resection, along with the biological features of the tumour, should influence the decision of the neurosurgeon to the patient follow up. PMID- 14740264 TI - Observation of arterial and venous thrombus formation by scanning and transmission electron microscopy. AB - BACKGROUND: In order to examine the process of thrombosis formation in artery and vein, the reactions of the arterial and venous endothelial surfaces were examined by scanning electron microscopy (SEM) and transmission electron microscopy (TEM) in the photothrombosis model. MATERIALS AND METHODS: Thirty-nine rats were divided into the following 4 groups: 1) Sham group (n = 5) without illumination, 2) group A (n = 10) irradiated for 1 min, 3) group B (n = 10) irradiated for 5 min, 4) group C (n = 14) irradiated for 10 min at the level of the left common carotid artery and internal jugular vein. RESULTS: SEM provided no evidence of damage or adhesion of blood platelets to the endothelium of either the artery or vein in shams or group A animals. In group B, evidence of damage to endothelial cell membrane (e.g., plasmalemmal pits, crater-like structures associated with tears between endothelial cells, and decreased number of microvilli) was obtained in the arterial wall but not in the vein. In group C, there was extensive or widespread adhesion of blood platelets and other cells, tears between arterial endothelial cells, and a decrease in the number of microvilli in the artery but not in the vein (p < 0.05). CONCLUSIONS: Cell membrane injuries, tears between the endothelial cells, and endothelial detachment occur before adhesion of blood platelets and thrombus formation in the blood vessel occlusion model by photochemical reaction. These changes occur significantly earlier in the artery than in the vein. PMID- 14740265 TI - Three-layer reconstruction with fascia lata and vascularized pericranium for anterior skull base defects. AB - BACKGROUND: We report an assessment of the efficiacy of a triple layer graft composed of fascia lata and vascularized pericranium for anterior skull base reconstruction. This technique is based on the concept that vascularized tissue over a free flap may promote vascularization and rapid wound healing. METHOD: A large fascial graft is prepared from the fascia lata and divided in two pieces and trimmed to a size larger than the bone and dural defect. Vascularized pericranium is harvested after bicoronal incision and elevating the bifrontal scalp flap down to the supraorbital rims. First is dural repair, which is performed with fascia lata placed between the brain and remaining dura. Second, fascia lata is placed over the skull base defect and secured with mini titanium screws over the cranial surface of the orbital ridges. Third, vascularized pericranium is laid between the two layers of fascia lata. FINDINGS: We studied 17 patients of whom 2 had malignancy, 6 had olfactory groove meningioma, 6 had skull base fracture and rhinorrhea, 1 case had orbital meningioma, 1 had invasive pituitary adenoma and 1 had basal encephalocele. The transbasal approach was used as a single procedure in 13 cases. The extended transbasal approach combined with a transfacial approach was used in 3 cases and with a pterional approach in 1 case. In each patient, reconstruction of the cranial base was performed with triple layer graft of fascia lata and vascularized pericranium. The patients were followed-up 2 months to 5 years. None of the patients experienced postoperative cerebrospinal fluid leakage, meningitis, abscess, brain herniation and tension pneumocephalus. INTERPRETATION: Fascia lata with vascularized pericranium is highly reliable, tensile and well suited for reconstruction of the anterior skull base. PMID- 14740266 TI - Development of malignant glioma 15 months after anterior temporal lobectomy in a patient with temporal lobe epilepsy. AB - We report a 36-year-old woman, who had previously undergone anterior temporal lobectomy for intractable temporal lobe seizures; fifteen months later, magnetic resonance (MR) images showed a space-occupying lesion in the temporal lobectomy cavity. After a second operation, a histopathological examination showed a grade III astrocytoma. The fortuitous co-occurrence of temporal lobe epilepsy and a tumour was suspected, but histopathological and immunohistochemical examination of original resected temporal lobe parenchyma did not show evidence of neoplasm. The patient had not undergone postoperative radiotherapy and had not experienced viral infections. We propose that two factors possibly associated with the development of glioma were chemical exposure from anticonvulsant agents and trauma from resection of the anterior temporal lobe during initial surgery. PMID- 14740267 TI - Primary-isolated optic nerve sarcoidosis. AB - Sarcoidosis is a systemic, idiopathic granulomatous disorder with occasionally surprising clinical presentations. A primary involvement of the optic nerve is particularly important due to visual prognosis. We report here a patient with occult sarcoidosis who presented to us with progressive visual loss as the first and primary manifestation of the disease. The patient underwent surgery for histopathological diagnosis and decompression of the optic nerve. This case demonstrated that sarcoidosis should be considered in the differential diagnosis of any lesion involving the optic nerve. PMID- 14740268 TI - Mixed meningeal and brain plasma-cell granuloma: an example of an unusual evolution. AB - We report a study of a 22-year-old woman with a plasma-cell granuloma (PCG), a rare intracranial lesion characterized by a non-neoplastic polyclonal proliferation of plasma cells and other mononuclear cells. She presented after a generalized seizure and CT-scan and magnetic resonance images demonstrated a left temporo-basal tumour mass involving both the meningeal layers and the brain parenchyma. Histopathological examination of a biopsy led to the diagnosis of a typical PCG. After a short course of steroid administration, the clinical and radiological features improved and complete regression of the lesion was shown after one year and persisted at four-year follow-up. This dramatic regression of an intracranial PCG shows that neither surgical removal nor radiation therapy is required to treat a broad skull base PCG. PMID- 14740269 TI - Primary multiple cerebral hydatid cysts with unusual features. AB - We present a case of 45 years old female patient with two primary cerebral hydatid cysts, each localised in separate hemispheres. Detailed studies revealed no evidence of hydatid disease elsewhere in the body. The cysts were removed by two separate operations. Though the cyst in the right posterior parietal lobe was delivered unruptured, seven months later a new cyst was detected in the same cavity, which seems to show relatively rapid growth in a year, and was also removed unruptured. This case is presented here for its unusual features such as having two primary cerebral hydatid cysts simultaneously in separate hemispheres, one of which partly localised intraventricularly, and the occurrence of a new cyst in the same cavity following intact removal of the previous cyst. PMID- 14740270 TI - Multiple basilar artery trunk aneurysms associated with fibromuscular dysplasia. AB - An association between fibromuscular dysplasia (FMD) and intracranial aneurysms has been proposed but the occurrence of multiple aneurysms on the trunk of basilar artery (BA) associated with FMD has not been previously reported. PMID- 14740271 TI - A glioma with an arteriovenous malformation: an association or a different entity? AB - A 58-year-old male was admitted with headache to our neurosurgery clinic. His neurological examination revealed slight left hemiparesis. The radiological evaluation with contrast administred magnetic resonance imaging (MRI) scan demonstrated a right temporo-parietal ring enhancing mass lesion surrounded by edema which was resembling a typical glioma (Fig. 1). The patient was operated on via a temporo-parietal craniotomy and an arteriovenous malformation surrounded by abnormal glial tissue was observed during the exposure. A nidus supplied by several branches arising from the middle cerebral artery (MCA) was obvious. The venous drainage of the malformation was to the superficial venous system. The observed arterial feeders and the draining vein were coagulated and the nidus was macroscopically totally excised. The frozen examination from surrounding glial tissue revealed a high grade glioma. The tumor was also macroscopically totally excised. Postoperatively, the cerebral angiogram demonstrated a right temporal arteriovenous malformation with a centrally excised nidus. The remaining major feeders involved the angular gyrus and the posterior temporal arteries. The venous drainage was to the straight and sigmoid sinuses (Fig. 2). The final histopathological examination of the specimen revealed an arteriovenous malformation surrounded by a high grade glioma (Fig. 3). The patient refused a second operation for total removal of the AVM. Postoperatively, he is doing well with improvement of his left hemiparesis. PMID- 14740272 TI - Trispan-assisted coiling of a wide-necked Persistent Trigeminal Artery aneurysm. PMID- 14740273 TI - Biochemical markers of types I and III collagen and limited joint mobility in type 1 diabetic patients. AB - Limited joint mobility (LJM), a long-term complication of diabetes, has been shown to be associated with microvascular complications of diabetes. Connective tissue alterations may contribute to the development of LJM and other diabetic complications. We tested whether biochemical markers of types I and III collagen metabolism are associated with LJM in type 1 diabetes. We studied 28 male patients of mean age 43.4 years (SD=9.5) and with a duration of diabetes of 25.2 years (SD=9.7) years. LJM assessment included goniometric measurements of the joints and classification by Rosenbloom's method. We measured serum concentrations of aminoterminal propeptide of type III procollagen (PIIINP), carboxyterminal propeptide of type I procollagen (PICP) and carboxyterminal crosslinked telopeptide of type I collagen (ICTP); urinary excretion of crosslinked N-telopeptides of type I collagen (NTX) and deoxypyridinoline crosslinks (DPyr) was also measured. Although average serum PIIINP tended to be higher in subjects with moderate-severe LJM (3.1 +/- 1.3 microg/l) than in subjects with mild LJM (2.5 +/- 0.7 microg/l) or without LJM (2.6 +/- 0.4 microg/l), no significant association was found (p<0.27). Concentrations of the other collagen markers were not different in subjects with or without LJM. We conclude that synthesis and degradation of types I and III collagen in diabetic subjects with LJM did not differ from those without LJM to reflect changes in the biochemical markers of these proteins. PMID- 14740274 TI - Equipotency of insulin glargine and regular human insulin on glucose disposal in healthy subjects following intravenous infusion. AB - The absolute glucose disposal of insulin glargine (Lantus) was compared to that of regular human insulin in healthy subjects (n=20) using the euglycaemic clamp technique in a single-dose, double-blind, randomized, two-way crossover design. Subjects received 30-minute intravenous infusions of insulin glargine (0.1 IU/kg) or human insulin (0.1 IU/kg) and a 20% glucose solution infused at a variable rate to maintain euglycaemia at the subject's baseline glucose level. At equal baseline blood glucose levels (4.42 mmol/l [range, 4.00-5.16 mmol/l] and 4.42 mmol/l [range, 4.01-4.94 mmol/l], respectively), the area under the glucose infusion rate (GIR) time curves from 0-6 hours (AUC(0-6h)) was within the bioequivalence range (insulin glargine, 663.92 mg/kg; human insulin, 734.85 mg/kg). Both the time to maximum GIR and the suppression of serum C-peptide were similar with insulin glargine and human insulin. The resulting maximum serum insulin concentrations (Cmax) were 151.16 microIU/ml and 202.23 microIU/ml, and the time to Cmax (Tmax) was 30 minutes (the duration of the infusion). The observed differences in the Cmax (the mean value for insulin glargine was about 25% lower than that of human insulin) could be explained by lower cross reactivity of insulin glargine in the human insulin radioimmunoassay. The employed intravenous route, though definitely not the intended clinical use of insulin glargine, provided the clinical evidence in healthy subjects that on a molar basis insulin glargine is equipotent to regular human insulin regarding glucose disposal. PMID- 14740275 TI - A 25-year follow-up study of glucose tolerance in first-degree relatives of type 2 diabetic patients: association of impaired or diabetic glucose tolerance with other components of the metabolic syndrome. AB - A follow-up study of first-degree relatives of type 2 diabetic patients presented the opportunity to study the association of components of the metabolic syndrome with oral glucose tolerance in these subjects. In 1992, 25 years after the first analysis of the cohort, we performed 75-g oral glucose tolerance tests and measured anthropometric data (body mass index, waist-hip ratio), insulin and C peptide concentrations, and parameters of lipoprotein metabolism (free fatty acids, triglycerides, cholesterol, HDL cholesterol). Of 135 participants, 71 had normal glucose tolerance (GT), 22 had impaired GT, and 42 had diabetic GT (WHO 1985 criteria). Impaired glucose tolerance and diabetes were significantly (Kruskal-Wallis test) associated with advanced age (p=0.001), higher body mass index (p=0.005) and waist-hip ratio (p=0.027), systolic hypertension (p=0.031), elevated basal insulin concentrations (p<0.001), higher free fatty acids (p<0.001) and triglycerides (p=0.017), and lower HDL cholesterol (p=0.003); no associations were found with total and LDL cholesterol levels (Friedewald's formula, p=0.25). Abnormalities (obesity, hypertriglyceridemia, low HDL cholesterol, hypertension, pathological oral glucose tolerance) were associated with significant deterioriations in all other components of the metabolic syndrome, if their number exceeded three. Disturbances of oral glucose tolerance are present in a high percentage of first-degree relatives after 25 years of follow-up (51% of those tested). Impaired or diabetic glucose tolerance in such a cohort was associated with overweight, hypertension and disturbances of lipoprotein metabolism characteristic of the metabolic syndrome. Hypercholesterolemia (LDL-cholesterol) is not a component of the metabolic syndrome in a German population with a high hereditary burden regarding type 2 diabetes. A metabolic syndrome should certainly be diagnosed if three components are present, although even in the presence of only two components, an elevated risk is evident. PMID- 14740276 TI - Patients' health education and diabetes control in a developing country. AB - We previously reported poor metabolic control in type 2 diabetic patients attending 2 primary care clinics in Trinidad. In an attempt to explain the poor metabolic control, we assessed primary care patients' theoretical knowledge of diabetes control and risk factors. Two hundred fifty-four diabetic out-patients recruited consecutively were asked by questionnaire: (i) if they were aware that family history of diabetes, obesity, physical inactivity and cigarette smoking were diabetes risk factors; (ii) if they knew the benefits of weight loss, exercise and healthy diet in diabetes management, and (iii) what where their common sources of diabetes health information. Although the majority of the patients (81.1%) were unaware that cigarette smoking is a diabetes risk factor, a majority were aware that obesity (66.3%), physical inactivity (73.5%) and being a relative of a diabetic patient (78.7%) constitute diabetes risk factors. Again, the majority of the patients were aware that healthy diet (94.9%), exercise (94.5%) and weight loss (87.4%) are beneficial in diabetes control. While media (48.6%) was the commonest source of diabetes information, doctors and nurses were consulted by 39.9% and 11.0% of patients, respectively. Type 2 diabetic patients in these clinics were well informed about diabetes risk factors and benefits of healthy lifestyle. Given our recent reports on poor metabolic control, application of this theoretical knowledge in controlling their diabetes remains doubtful. PMID- 14740277 TI - Silent coronary artery disease in patients with type 2 diabetes mellitus. AB - The purpose of this study was to estimate the prevalence and risk factors of silent CAD in asymptomatic type 2 diabetic patients aged over 40 years. A total of 172 asymptomatic type 2 diabetic patients, mean age 54.42 years, with normal resting electrocardiogram were included in the study. Technetium-99m (Tc-99m) tetrofosmin cardiac single photon emission computed tomography myocardial scintigraphy with exercise testing or dipyridamole injection was performed on all patients. If this test was positive, coronary angiography was carried out and was considered to be positive with a stenosis of > or =70%. Abnormal perfusion pattern was found in 14 patients (8.14%). Significant coronary artery stenosis was found in 13 subjects (7.56%), confirming a high positive predictive value (92.86%) of this diagnostic procedure. A significant correlation was observed between silent CAD and male sex, retinopathy, hypertension, post-prandial blood glucose level, and low HDL-cholesterol level. Sex (OR=4.026; 95% CI, 1.187 13.659), hypertension (OR=5.564; 95% CI, 1.446-21.400) and retinopathy (OR=3.766; 95% CI, 1.096-12.948) were risk factors for CAD. Overall, 14.06% of asymptomatic male patients with type 2 diabetes mellitus presented silent CAD with significant angiographically documented coronary stenosis. This finding, along with the high positive predictive value of a noninvasive technique, indicates that routine screening for silent CAD would be useful in this patient subgroup especially when they have retinopathy or hypertension. PMID- 14740278 TI - Fasting plasma glucose and glycated haemoglobin in the screening of diabetes and impaired glucose tolerance. AB - The use of fasting plasma glucose (FPG) only has been proposed for the screening and diagnosis of diabetes, but its sensitivity has been reported to be unsatisfactory. The use of HbA1C, alone or combined with FPG, has been suggested for the screening of diabetes and impaired glucose tolerance (IGT). In a sample of 1215 adult subjects without previously known diabetes, we assessed the sensitivity and specificity of FPG and HbA1C in diagnosing diabetes and IGT, determined by oral glucose tolerance test (OGTT). All lean diabetic patients, and 85% of overweight and obese diabetic individuals, had FPG > or =7 mmol/l. FPG >6.1 mmol/l had a sensitivity of 98.8% and a specificity of 32.9%; HbA1C had a lower specificity and sensitivity for the screening of diabetes. A screening strategy for diabetes based on FPG, with OGTT in all overweight subjects with FPG >6.1 mmol/l, is suggested. Neither FPG nor HbA1C is effective in the screening of IGT; although combined FPG and HbA1C could be useful for case finding, screening for IGT with OGTT is advisable in all subjects at high risk. PMID- 14740279 TI - Importance of premeal injection time in insulin therapy: Humalog Mix25 is convenient for improved post-prandial glycemic control in type 2 diabetic patients with Italian dietary habits. AB - We investigated the use, in a short period, of Humalog Mix25 (Mix25) in a twice daily administration regimen compared to a twice-daily injection therapy with Humulin 30/70 (30/70) in diabetic patients with Italian dietary habits. We studied 33 type 2 diabetic patients aged 59.1 +/- 8.1 years, BMI 29.8 +/- 2.7 kg/m2, duration of diabetes and insulin therapy of 14.4 +/- 9.8 and 4.2 +/- 4.6 years, respectively. After a 4-day lead-in period of twice-daily human insulin 30/70 treatment, patients were randomized to one of two treatment sequences: (1) a twice-daily regimen with Mix25 just 5 minutes before the morning and evening meals for 12 days, followed by a twice-daily therapy with human insulin 30/70 given 30 minutes before the morning and evening meals for an additional 12 days; or (2) the alternate sequence. Each patient underwent a mixed meal test: Humulin 30/70 was administered 30 minutes before the meal, while Mix25 was given 5 minutes before. The 2-hour post-prandial glucose concentration after breakfast was significantly lower during treatment with Mix25 than with Humulin 30/70 (157 +/- 43.2 vs. 180 +/- 43.2 mg/dl, p<0.05). The glycemic excursion after dinner on Mix25 treatment was significantly lower than with Humulin 30/70 (12.2 +/- 48.01 vs. 35.5 +/- 36.92 mg/dl, p<0.05). AUCglucose after Mix25 was lower than after Humulin 30/70. Glycemia after test meal was significantly lower with Mix25 than with Humulin 30/70. Insulin and free insulin concentrations after the test meal were significantly higher with Mix25 in comparison to Humulin 30/70. AUC serum insulin and free insulin curves after Mix25 were significantly higher than after Humulin 30/70 (p=0.028 and p=0.005, respectively). Twice-daily injections of Humalog Mix25, compared to human insulin 30/70 in type 2 diabetic patients with Italian dietary habits, provide improved and lasting post-prandial glycemic control, with the great convenience of the injection just before the meal. PMID- 14740281 TI - Is a specific oncological scoring system better at predicting the prognosis of cancer patients admitted for an acute medical complication in an intensive care unit than general gravity scores? AB - OBJECTIVE: To evaluate the effectiveness of a specific oncologic scoring system the ICU Cancer Mortality model (ICM)-in predicting hospital mortality in comparison to two general severity scores-the Acute Physiology and Chronic Health Evaluation (APACHE II) and the Simplified Acute Physiology Score (SAPS II). PATIENTS AND METHODS: All 247 patients admitted for a medical acute complication over an 18-month period in an oncological medical intensive care unit were prospectively registered. Their data, including type of complication, vital status at discharge and cancer characteristics as well as other variables necessary to calculate the three scoring systems were retrospectively assessed. RESULTS: Observed in-hospital mortality was 34%. The predicted in-hospital mortality rate for APACHE II was 32%; SAPS II, 24%; and ICM, 28%. The goodness of fit was inadequate except for the ICM score. Comparison of the area under the ROC curves revealed a better fit for ICM (area 0.79). The maximum correct classification rate was 72% for APACHE II, 74% for SAPS II and 77% for ICM. APACHE II and SAPS II were better at predicting outcome for survivors to hospital discharge, although ICM was better for non-survivors. Two variables were independently predicting the risk of death during hospitalisation: ICM (OR=2.31) and SAPS II (OR=1.05). CONCLUSIONS: Gravity scores were the single independent predictors for hospital mortality, and ICM was equivalent to APACHE II and SAPS II. PMID- 14740282 TI - Reliability and validity of the Functional Assessment of Chronic Illness Therapy Spiritual (FACIT-Sp) for Japanese patients with cancer. AB - The reliability and validity of the Japanese version of the Functional Assessment of Chronic Illness Therapy-Spiritual (FACIT-Sp) scale were assessed. This scale was developed in the United States to assess quality of life (QOL) in relation to spirituality. Two surveys were conducted on each of 306 cancer patients. In addition to the FACIT-Sp, the Hospital Anxiety and Depression Scale (HADS) was administered. Cronbach's alpha reliability coefficient, an evaluation of internal consistency, for the FACIT-Sp subscales ranged from 0.81 to 0.91. There were no significant differences between the patients evaluated using the FACIT-Sp subscale and the HADS subscale with regard to degree of religious feelings. The correlation coefficients between the FACIT-Sp and the HADS depression and anxiety scales indicated a moderate correlation. These findings suggest that the Japanese version of the FACIT-Sp scale is satisfactory in terms of reliability and validity and is a useful tool in the study of spirituality among Japanese cancer patients. PMID- 14740283 TI - Darbepoetin alfa in lung cancer patients on chemotherapy: a retrospective comparison of outcomes in patients with mild versus moderate-to-severe anaemia at baseline. AB - GOALS: Currently, there is some debate concerning the haemoglobin level at which treatment of anaemia with erythropoiesis-stimulating agents should be initiated in cancer patients on chemotherapy. We report several analyses of data from a phase III trial of darbepoetin alfa versus placebo, comparing outcomes for patients with mild and moderate-to-severe anaemia. PATIENTS AND METHODS: Data were obtained from a phase III trial of darbepoetin alfa versus placebo in anaemic patients with lung cancer receiving chemotherapy ( n=314). Outcomes were compared for patients with baseline haemoglobin > or =10-11 g/dl and <10 g/dl. RESULTS: Darbepoetin alfa significantly reduced transfusions compared with placebo, irrespective of haemoglobin level at treatment initiation. For patients with baseline haemoglobin <10 g/dl, 31% and 59% of those receiving darbepoetin alfa and placebo, respectively, required a transfusion from week 5 to the end of the treatment phase ( P<0.038). For patients with baseline haemoglobin > or =10 g/dl, the proportions were 15% and 41%, respectively ( P<0.001). Darbepoetin alfa also improved fatigue compared with placebo in both haemoglobin categories. CONCLUSIONS: These findings show that initiating treatment at haemoglobin levels both <10 g/dl and > or =10-11 g/dl results in substantial clinical benefits, supporting the use of erythropoietic therapy also in patients with mild anaemia. PMID- 14740284 TI - A boy with consecutive development of SLE and Wegener granulomatosis. AB - An 11-year-old boy with consecutive development of systemic lupus erythematosus (SLE) and Wegener granulomatosis (WG) is presented. He was first admitted to the hospital with the findings of SLE, including crescentic glomerulonephritis, Coombs' test-positive hemolytic anemia, hypocomplementemia, antinuclear antibody (ANA) positivity, and elevated levels of anti-double-stranded (ds) DNA antibodies. He was treated successfully with steroids, cyclophosphamide, and peritoneal dialysis. One month after his discharge he developed an apparent viral infection. Three weeks afterwards he was readmitted with the findings of lower respiratory tract involvement, maxillary sinusitis, nasal septum perforation, p- and c-antineutrophil cytoplasmic antibody (ANCA) positivity, but normal complement, ANA, and anti-ds DNA levels, suggesting the diagnosis of WG. He did not respond to anti-infectious and immunosuppressive treatment, and he died of Pseudomonas sepsis. PMID- 14740285 TI - Effect of oral camostat mesilate on hematuria and/or proteinuria in children. AB - The effects of camostat mesilate (CM), a derivative of gabexate mesilate developed for oral use, on primary glomerulonephritis (GN) and chance hematuria and/or proteinuria were evaluated. Fourteen patients (6 males, 8 females) with a mean age of 11 years and 3 months (range 4-16 years) were enrolled. Histological and clinical diagnoses of the 14 patients were as follows: IgA nephropathy 3, non IgA GN 2, and asymptomatic significant microscopic hematuria [more than 100 red blood cells per high-power field (x400)] with or without proteinuria 9. They were consecutively treated with oral CM (100 mg twice a day) when they were confirmed to have continuous significant microscopic hematuria and/or proteinuria after a few months of observational follow-up. Urinary findings were normalized in 10 of the 14 patients (85.7%) between 1 month 1 week and 10 months (mean 4 months) after administration of CM. Hematuria cleared in 11 of 13 patients, and proteinuria disappeared in 4 of 5 patients. The mean duration of CM administration was 21.7 +/- 9.1 months (range 4-37 months). At present, 3-12 years after discontinuation of CM therapy, their urinary findings remain normal at 9 years 10 months to 26 years 6 months of age. In conclusion, there appears to be an association between the oral use of CM and reduction in significant microscopic hematuria and/or proteinuria. Oral CM therapy could represent a practical primary care approach to chance hematuria and/or proteinuria in children. PMID- 14740286 TI - Hemichorea with antiphospholipid antibodies in a patient with lupus nephritis. AB - Although central nervous system involvement is an important manifestation of systemic lupus erythematosus (SLE), chorea is a relatively uncommon complication. A strong association between chorea and the presence of antiphospholipid antibodies (aPLs) has been reported in patients with SLE, lupus-like disease, or primary antiphospholipid syndrome. We describe a patient with lupus nephritis and cerebral infarction, who subsequently developed recurrent hemichorea associated with increased aPLs levels. A 7-year-old boy suffered from lupus nephritis and a left middle cerebral artery infarction associated with aPLs. He subsequently experienced two episodes of right hemichorea associated with increased aPLs levels without any evidence of further neurological lesions by brain computed tomography or magnetic resonance imaging. The previous left cerebral artery infarction might have increased the susceptibility of the left basal ganglia to the effects of aPLs that contributed to the development of the right hemichorea in this patient. PMID- 14740287 TI - Solar UV-B radiation affects leaf quality and insect herbivory in the southern beech tree Nothofagus antarctica. AB - We examined the effects of solar ultraviolet-B (UV-B) radiation on plant-insect interactions in Tierra del Fuego (55 degrees S), Argentina, an area strongly affected by ozone depletion because of its proximity to Antarctica. Solar UV-B under Nothofagus antarctica branches was manipulated using a polyester plastic film to attenuate UV-B (uvb-) and an Aclar film to provide near-ambient UV-B (uvb+). The plastic films were placed on both north-facing (i.e., high solar radiation in the Southern Hemisphere) and south-facing branches. Insects consumed 40% less leaf area from north- than from south-facing branches, and at least 30% less area from uvb+ branches than from uvb- branches. The reduced herbivory on leaves from uvb+ branches occurred for both branch orientations. Leaf mass per area increased and relative water content decreased on north- versus south-facing branches, while no differences were apparent between the UV-B treatments. Solar UV-B did lead to lower gallic acid concentration and higher flavonoid aglycone concentration in uvb+ leaves relative to uvb- leaves. Both the flavonoid aglycone and quercetin-3-arabinopyranoside were higher on north-facing branches. In laboratory preference experiments, larvae of the dominant insect in the natural community, Geometridae "Brown" (Lepidoptera), consumed less area from field-grown uvb+ leaves than from uvb- leaves in 1996-97, but not in 1997-98. Correlation analyses suggested that the reduction in insect herbivory in the field under solar UV-B may be mediated in part by the UV-B effects on gallic acid and flavonoid aglycone. PMID- 14740288 TI - Seed dispersal by white-tailed deer: implications for long-distance dispersal, invasion, and migration of plants in eastern North America. AB - For many plant species in eastern North America, short observed seed dispersal distances (ranging up to a few tens of meters) fail to explain rapid rates of invasion and migration. This discrepancy points to a substantial gap in our knowledge of the mechanisms by which seeds are dispersed long distances. We investigated the potential for white-tailed deer ( Odocoileus virginianus Zimm.), the dominant large herbivore in much of eastern North America, to disperse seeds via endozoochory. This is the first comprehensive study of seed dispersal by white-tailed deer, despite a vast body of research on other aspects of their ecology. More than 70 plant species germinated from deer feces collected over a 1 year period in central New York State, USA. Viable seeds included native and alien herbs, shrubs, and trees, including several invasive introduced species, from the full range of habitat types in the local flora. A mean of >30 seeds germinated per fecal pellet group, and seeds were dispersed during all months of the year. A wide variety of presumed dispersal modes were represented (endo- and exozoochory, wind, ballistic, ant, and unassisted). The majority were species with small-seeded fruits having no obvious adaptations for dispersal, underscoring the difficulty of inferring dispersal ability from diaspore morphology. Due to their broad diet, wide-ranging movements, and relatively long gut retention times, white-tailed deer have tremendous potential for effecting long-distance seed dispersal via ingestion and defecation. We conclude that white tailed deer represent a significant and previously unappreciated vector of seed dispersal across the North American landscape, probably contributing an important long-distance component to the seed shadows of hundreds of plant species, and providing a mechanism to help explain rapid rates of plant migration. PMID- 14740289 TI - Decomposition rate of organic substrates in relation to the species diversity of soil saprophytic fungi. AB - Despite the great interest concerning the relationship between species diversity and ecosystem functioning, there is virtually no knowledge as to how the diversity of decomposer microbes influences the decomposition rate of soil organic matter. We established a microcosm study in which the number of soil fungi was investigated in relation to the system's ability to (i) degrade raw coniferous forest humus, and (ii) use resources that were either added to the systems or released into the soils after a disturbance (drought). With the exception of the most diverse treatment, in each of the six replicates of each of the six diversity treatments (1, 3, 6, 12, 24 or 43 taxa), fungal taxa were randomly chosen from a pool of 43 commonly isolated fungal species of raw humus. Two months after initiation of the study CO2 production increased as fungal diversity increased, but in the species-poor end of the diversity gradient only. Addition of various energy resources to the microcosms generally increased the level of soil respiration but did not affect the shape of the diversity-CO2 production curve. Rewetting the soil after severe drought resulted in a rapid flush of CO2, particularly in the most diverse communities. The biomass of the fungi in the non-disturbed soils, and soil NH4-N concentration and soil pH in both disturbed and non-disturbed systems were slightly but significantly higher in the diverse than in the simple systems. Fungal species richness had no influence on the organic matter content of the humus at the end of the experiment. The results suggest that the functional efficiency of fungal communities can increase with the number of fungal taxa. This diversity effect was, however, significant at the species-poor end of the diversity gradient only, which implies considerable functional equivalency (redundancy) among the decomposer fungi. PMID- 14740290 TI - Effects of dung and seed size on secondary dispersal, seed predation, and seedling establishment of rain forest trees. AB - Seeds dispersed by tropical, arboreal mammals are usually deposited singly and without dung or in clumps of fecal material. After dispersal through defecation by mammals, most seeds are secondarily dispersed by dung beetles or consumed by rodents. These post-dispersal, plant-animal interactions are likely to interact themselves, as seeds buried by dung beetles are less likely to be found by rodents than unburied seeds. In a series of three experiments with seeds of 15 species in central Amazonia (Brazil), we determined (1) how presence and amount of dung associated with seeds influences long-term seed fate and seedling establishment, (2) how deeply dung beetles bury seeds and how burial depth affects seedling establishment, and (3) how seed size affects the interaction between seeds, dung beetles, and rodents. Our overall goal was to understand how post-dispersal plant-animal interactions determine the link between primary seed dispersal and seedling establishment. On average, 43% of seeds surrounded by dung were buried by dung beetles, compared to 0% of seeds not surrounded by dung ( n=2,156). Seeds in dung, however, tended to be more prone than bare seeds to predation by rodents. Of seeds in dung, probability of burial was negatively related to seed size and positively related to amount of dung. Burial of seeds decreased the probability of seed predation by rodents three-fold, and increased the probability of seedling establishment two-fold. Mean burial depth was 4 cm (0.5-20 cm) and was not related to seed size, contrary to previous studies. Probability of seedling establishment was negatively correlated with burial depth and not related to seed size at 5 or 10 cm depths. These results illustrate a complex web of interactions among dung beetles, rodents, and dispersed seeds. These interactions affect the probability of seedling establishment and are themselves strongly tied to how seeds are deposited by primary dispersers. More generally, our results emphasize the importance of looking beyond a single type of plant-animal interaction (e.g., seed dispersal or seed predation) to incorporate potential effects of interacting interactions. PMID- 14740291 TI - Long-term effects of defoliation on quaking aspen in relation to genotype and nutrient availability: plant growth, phytochemistry and insect performance. AB - This research tested the long-term effects of defoliation on aspen chemistry and growth in relation to genotype and nutrient availability. We grew saplings of four aspen genotypes in a common garden under two conditions of nutrient availability, and subsequently subjected them to two levels of artificial defoliation. Artificial defoliation suppressed plant growth, and saplings of the four genotypes did not show evidence of genetic variation in tolerance to defoliation. Phenolic glycoside concentrations did not respond to defoliation, but were influenced by genotype and nutrient availability. Condensed tannins responded to defoliation and varied among genotypes. Although defoliation affected condensed tannins, plant quality was not altered in a manner important for gypsy moth performance. Regression analyses suggested that phenolic glycoside concentrations accounted for most of the variation in insect performance. The lack of a strong response important for herbivores was surprising given the severity of the defoliation treatment (nearly 100% of leaf area was removed). In this study, plant genotype was of primary importance, nutrient availability was of secondary importance and long-term induced responses were unimportant as determinants of insect performance. PMID- 14740292 TI - Delineating the effects of a plant trait on interactions among associated insects. AB - Plant traits can affect ecological interactions between plants, herbivores, and predators. Our study tests whether reduced leaf wax in peas alters the interaction between the pea aphid ( Acyrthosiphon pisum), a foliar-foraging predator (a lady beetle, Hippodamia convergens) and a ground-foraging predator (a ground beetle, Poecilus scitulus). We performed a 2 x 2 x 2 factorial experiment in which wax level, presence of H. convergens, and presence of P. scitulus were manipulated. Experimental arenas consisted of a cage surrounding three pea plants. One plant in each cage was stocked with 15 pea aphids. In greenhouse and field cage experiments, we assessed the effect of each factor and their interactions on aphid density. As in previous studies, H. convergens foraged for aphids more effectively on reduced wax peas than on normal peas. Other interactions among H. convergens, P. scitulus, and A. pisum were the same on both types of peas. We consider how aphid movement, plant growth, and a high frequency of predation by P. scitulus on H. convergens influenced pea aphid density. PMID- 14740293 TI - Platelet-derived growth factor exerts disparate effects on odontoblast differentiation depending on the dimers in rat dental pulp cells. AB - Platelet-derived growth factor (PDGF) has recently been demonstrated to control the expression of alkaline phosphatase and proteoglycan synthesis of odontoblastic cells in dental pulp tissues. Although PDGF appears to be closely related to dentinogenesis, much about the mode of action of PDGF on odontoblast differentiation remains unclear. In this study, we examined the effects of three PDGF dimers (PDGF AA, AB, and BB) on odontoblastic differentiation of dental pulp cells in long-term mineralized cultures. Dental pulp cells isolated from rat lower incisors were continuously treated with each of PDGF AA, AB, and BB in separate cultures for 20 days. The three PDGF dimers suppressed alkaline phosphatase activity, osteocalcin and calcium content, and the formation of dentin-like nodules. The expression of mRNA for dentin sialoprotein (DSP) in the cells was inhibited by PDGF AA treatment, whereas PDGF AB and BB treatment stimulated the expression of DSP, even though the dentin-like nodule formation was inhibited. Although the effects of PDGF on odontoblastic differentiation varied among the dimers, the cells expressed both PDGF alpha and beta receptors, whose quantities were similar. These results suggest that PDGF exerts diverse effects on odontoblastic differentiation depending on its dimeric form. These in vitro findings explain, at least in part, the in vivo action of PDGF in dentinogenesis during the repair process of damaged dental pulp. PMID- 14740294 TI - Contrasting patterns of Y chromosome variation in Ashkenazi Jewish and host non Jewish European populations. AB - The molecular basis of more than 25 genetic diseases has been described in Ashkenazi Jewish populations. Most of these diseases are characterized by one or two major founder mutations that are present in the Ashkenazi population at elevated frequencies. One explanation for this preponderance of recessive diseases is accentuated genetic drift resulting from a series of dispersals to and within Europe, endogamy, and/or recent rapid population growth. However, a clear picture of the manner in which neutral genetic variation has been affected by such a demographic history has not yet emerged. We have examined a set of 32 binary markers (single nucleotide polymorphisms; SNPs) and 10 microsatellites on the non-recombining portion of the Y chromosome (NRY) to investigate the ways in which patterns of variation differ between Ashkenazi Jewish and their non-Jewish host populations in Europe. This set of SNPs defines a total of 20 NRY haplogroups in these populations, at least four of which are likely to have been part of the ancestral Ashkenazi gene pool in the Near East, and at least three of which may have introgressed to some degree into Ashkenazi populations after their dispersal to Europe. It is striking that whereas Ashkenazi populations are genetically more diverse at both the SNP and STR level compared with their European non-Jewish counterparts, they have greatly reduced within-haplogroup STR variability, especially in those founder haplogroups that migrated from the Near East. This contrasting pattern of diversity in Ashkenazi populations is evidence for a reduction in male effective population size, possibly resulting from a series of founder events and high rates of endogamy within Europe. This reduced effective population size may explain the high incidence of founder disease mutations despite overall high levels of NRY diversity. PMID- 14740295 TI - Patterns of linkage disequilibrium in the MHC region on human chromosome 6p. AB - Single nucleotide polymorphisms (SNPs) in the human genome are thought to be organised into blocks of high internal linkage disequilibrium (LD), separated by intermittent recombination hotspots. Since understanding haplotype structure is critical for an accurate assessment of inter-individual genetic differences, we investigated up to 968 SNPs from a 10-Mb region on chromosome 6p21, including the human major histocompatibility complex (MHC), in five different population samples (45-550 individuals). Regions of well-defined block structure were found to coexist alongside large areas lacking any clear structure; occasional long range LD was observed in all five samples. The four white populations analysed were remarkably similar in terms of the extend and spatial distribution of local LD. In US African Americans, the distribution of LD was similar to that in the white populations but the observed haplotype diversity was higher. The existence of large regions without any clear block structure renders the systematic and thorough construction of SNP haplotype maps a crucial prerequisite for disease association studies. PMID- 14740296 TI - TP53 haplotype-based analysis and incidence of post-angioplasty restenosis. AB - The tumor suppressor gene product, in particular tumor suppressor protein p53 (TP53), has been suggested to play a role in post-angioplasty restenosis. However, no genetic-epidemiological studies relating to TP53 gene polymorphism(s) and the incidence of post-angioplasty restenosis are available. TP53 11951_11966dup16bp, R72P, and 13494G>A polymorphisms were characterized in a cohort of 779 patients, of whom 342 cases had developed restenosis (as defined by >50% loss of lumen compared with immediate post-procedure results) at repeat quantitative coronary angiography at six months post angioplasty. The haplotype frequency distribution was marginally different between cases and controls with restenosis risk (chi(2)(7df)=13.08, P=0.070). Multivariable haplotype-based logistic regression indicated that haplotypes 16bp(-) -P72-G13494 [corrected], and 16bp(+) -P72-A13494 [corrected] exhibit protective effects on restenosis risk (odds ratio=0.58, 95%CI=0.40-0.83, P=0.0033; odds ratio=0.69, 95%CI=0.48-0.99, P=0.049, respectively). Multivariable haplotype-based linear regression again showed similar, significant association with degree of lumen loss. The present findings indicate protective effects of TP53 16bp(-) -P72-G13494 [corrected], and 16bp(+) -P72-A13494 [corrected] haplotypes in the incidence of restenosis after angioplasty. Furthermore, our study demonstrates that a haplotype-based approach can be more informative than a single-marker or marker-by-marker analysis. PMID- 14740297 TI - Cloning, expression and characterization of a lipase gene (lip3) from Pseudomonas aeruginosa LST-03. AB - A lipase gene (lip3) was cloned from the Pseudomonas aeruginosa strain LST-03 (which tolerates organic solvents) and expressed in Escherichia coli. The cloned sequence includes an ORF consisting of 945 nucleotides, encoding a protein of 315 amino acids (Lip3 lipase, 34.8 kDa). The predicted Lip3 lipase belongs to the class of serine hydrolases; the catalytic triad consists of the residues Ser-137, Asp-258, and His-286. The gene cloned in the present study does not encode the LST-03 lipase, a previously isolated solvent-stable lipase secreted by P. aeruginosa LST-03, because the N-terminal amino acid sequence of the Lip3 lipase differs from that of the LST-03 lipase. Although the effects of pH on the activity and stability of the Lip3 lipase, and the temperature optimum of the enzyme, were similar to those of the LST-03 lipase, the relative activity of the Lip3 lipase at lower temperatures (0-35 degrees C) was higher than that of the LST-03 lipase. In the absence of organic solvents, the half-life of the Lip3 lipase was similar to that of the LST-03 lipase. However, in the presence of most of the organic solvents tested in this study (the exceptions were ethylene glycol and glycerol), the stability of the Lip3 lipase was lower than that of the LST-03 lipase. PMID- 14740299 TI - [Predictors and mechanisms of tinnitus distress - a longitudinal analysis]. AB - BACKGROUND: The available cross-sectional and retrospective evidence does not provide a clear answer to the question whether the multiple psychological problems and disturbances found in patients with chronic decompensated tinnitus are cause or consequence of the tinnitus. The present research took a longitudinal approach to examine this question. METHOD: Psychopathological symptoms, personality, socio-demographical variables and otological features were assessed in 48 patients which had suffered from tinnitus for a period of no longer than 4 weeks. Six months after first assessment 92 % of the original sample (n = 44) were re-examined. 34 of these patients displayed chronic tinnitus. Univariate and multiple regression analyses were conducted in order to identify variables at first assessment which predicted tinnitus distress at second assessment for the 34 patients with chronic tinnitus. RESULTS: Tinnitus attributed sleep disturbance, anxiousness, and life satisfaction, each assessed at first investigation, independently predicted tinnitus distress at second assessment six months later. These three variables together predicted 56 % of the variance of tinnitus distress at second assessment. CONCLUSIONS: Our results support the model that the symptom tinnitus may develop on the basis of an enhanced psychophysiological tension and become a condensational core of preexisting psychological distress as a facilitatory process. Our results suggest that early psychotherapeutic interventions in patients at risk may prevent decompensation. PMID- 14740300 TI - [Expression of osteoclast stimulating and differentiating factors in a murine model of localized inflammatory bone resorption]. AB - BACKGROUND: The pathology associated to cholesteatoma is predominantly a consequence of osteoclast-mediated bone resorption within the middle ear. To assess its pathogenesis a murine model for dermal-implant induced osteolysis was evaluated for the expression of osteoclast stimulating and differentiating factors. METHODS: Mouse calvaria were analysed for the expression of osteoprotegerin ligand (OPGL), osteoprotegerin (OPG) and macrophage-colony stimulating factor (M-CSF) using immunohistochemistry. The detection of osteoclast cell lineage was acquired by immunohistochemistry using markers CD 4, CD 11a, CD 11b, CD 14, CD 51, CD 68 and TRAP. RESULTS: An increased expression of the investigated cytokines M-CSF, OPG and OPGL was demonstrated by immunohistochemistry. The presence of osteoclast precursor cells and mature resorbing osteoclasts was confirmed in time-dependent manner triggered by dermal implantation. CONCLUSIONS: This study reveals the basic events in osteoclast biology in localized inflammatory bone resorption and provides new insights into the comprehension of cholesteatoma-induced bone resorption. PMID- 14740301 TI - [Bilateral cochlea implantation in varying duration of deafness]. AB - The effectiveness of cochlear implants in profoundly deaf patients could be demonstrated in the last years. One of the problems of these patients is the decreased speech understanding in noise. Therefore bilateral cochlear implantation seems to be obvious. Another problem of unilateral cochlear implantation is to predict the better side for implantation concerning speech understanding. Only a few factors seem to help for the choice of the side for implantation. One of these is duration of deafness. In other studies short duration of deafness is a significant predictor of good word recognition. We present a case of a deaf patient with a time gap of 23 years for deafness between both ears. The patient underwent bilateral cochlear implantation (Med-el Combi 40+) as one stage procedure. After activation of the speech processor he shows better speech understanding on that ear, which is deaf for more than 25 years. In addition he has further improvement of speech understanding in noise by using both implants compared to one. In unilateral cochlear implantation selection of the side might still be difficult. In bilateral cochlear implantation the better side is always implanted, which is an other advantage of bilateral cochlear implantation. PMID- 14740302 TI - [Complications after endonasal surgery of the paranasal sinuses for inflammatory diseases]. AB - BACKGROUND: There are sufficient reports on the incidence of complications after endonasal surgery of the paranasal sinuses. It is the aim of this study to calculate the incidence of typical complications after primary and revision surgery, after "functional" and "extended" operations, together with the influence of expertise and of different optical devices on the complications rate. Preoperative computer tomography of the paranasal sinuses are the golden standard. There exists a historical interest to compare the complication rate in patients operated without and those with a CT, especially in those countries where the availability of CTs is low or not existent. METHODS: 266 consecutive patients who underwent an endonasal surgery of the paranasal sinuses from January 1989 to December 1990 were revised to study the incidence and relative risk rate of typical complications after primary and revision surgery, after "functional" or "extended" surgery, the rate for more experienced or less experienced surgeons, as well as the differences between endoscopic or microscopic surgery. The study also compares the complication rate of patients operated on with and without a preoperative computed tomography. RESULTS: 207 out of 263 patients (78.7 %) had no complication. 58 minor and three severe complication were registered for 56 patients. 6 patients had two or more minor complications. No surgeon showed a higher complication rate compared to others (p = 0.33), and there was no statistically significant difference (p = 0.279) in the complication rate between experienced and less experienced surgeons. Endoscopic or microscopic surgery showed no statistically significant differences (p = 0.117) and extended sinus surgery displayed more complications compared to minor surgery (p = 0.041), as well as primary surgery compared to revision surgery (p = 0.003). No statistically significant differences could be found for surgeries performed with or without computed tomography (p = 0.097). CONCLUSIONS: A higher complication rate is to be expected for primary surgery and after extended sinus surgery, especially when an infundibulotomy was performed. In our series no learning curve was observed, especially as the variable "experience" showed no statistically significant difference. Endonasal surgery without CAT scan is not recommended, however, no higher complication rate was found in surgery without CAT scan. PMID- 14740303 TI - [Management of nasal fistulas with intracranial extension]. AB - BACKGROUND: Medial nasal fistulas are rare congenital malformations that mostly become obvious by recurrent infections such as secretion from the fistula opening or swelling of the nasal tissues. In rare cases, these fistulas may extend intracranially with meningeal irritations. PATIENT PRESENTATION: Three patients with medial nasal fistulas were operated during the last two years in our department. In two children, the fistulas ended at the glabella. In one older boy, the fistula revealed an intracranial extension. In the last case, a combined rhino- and neurosurgical approach was applied to perform a complete resection of the fistula. DISCUSSION: During the second embryonic month of facial development, the medial nasal processes approach each other with a fusion of their epithelial covering that completely disappears during later stages. If parts of this epithelial fusion zone persist, epithelial remnants may develop into nasal fistulas or dermoids. The adequate therapy of these nasal malformations consists of a complete surgical removal. PMID- 14740304 TI - [Malignomas of the nasal cavity and the paranasal sinuses: clinical characteristics, therapy and prognosis of different tumor types]. AB - BACKGROUND: Malignomas of the nasal cavity and the paranasal sinuses count for less than 3 % of the ENT-malignancies. PATIENTS AND METHODS: This retrospective chart review reports about 46 patients that were first diagnosed and treated with a nasal cavity or paranasal sinus malignancy between 1998 and 2002 in the ENT department of the Ruhr-Universitat Bochum. RESULTS: 18 of 46 malignomas were squamous cell carcinomas. At initial diagnose, already 52 % of malignomas were staged T4. Accordingly the origin in 17 % of malignomas could not be determined anymore. Additional 43 % of malignomas originated from the nasal cavity, another 20 % from the maxillary sinus. 11 of 46 patients (24 %) showed histological proven nodal involvement at initial diagnose. 92 % of patients underwent surgical therapy. 25 patients were radiated postoperatively. Over-all 1-year-survival-rate was 75 %, over-all 2-year-survival-rate was 47 %. Beside patients age, tumor free margins at initial surgery and absence of nodal involvement correlated to improved survival. 16 (76 %) of the 21 patients who died had local recurrence. Maxillary sinus malignomas recurred more often than malignomas of the ethmoid and the nasal cavity. CONCLUSIONS: Malignancies of the nasal cavity and paranasal sinuses are very often diagnosed in advanced T-stages because of unspecific symptoms. The limited prognosis mainly depends on free surgical margins at the first resection and nodal involvement at the first diagnose. For improvement in outcome of nasal and paranasal sinuses malignancies, prospective multi-center trials are necessary. PMID- 14740305 TI - [Dacryocystorhinostomy - state of the art, indications, results]. AB - BACKGROUND: In the ophthalmological literature external dacryocystorhinostomy (DCR) is considered the gold standard for the treatment of lacrimal duct stenoses. Rhinologists, on the other hand, favour the endonasal approach. MATERIALS AND METHODS: On the basis of an extensive review of the literature and our own longstanding experience we present an overview of the causes, the necessary diagnostic procedures and the surgical management of lacrimal duct stenosis. The outcomes of the two operative approaches are compared and special operative techniques and aids such as the use of laser, endoscopic dacryoplasty, silicone stenting, silicone cones with lacrimal duct stenting and the use of mitomycin C are discussed. RESULTS: The results published for endonasal DCR are slightly worse than those for the external operation. The success rates are around 90 %. Laser-assisted DCR and endoscopic dacryoplasty do not currently appear to yield better results than the conventional methods. Silicone stenting is not necessary in conventional endonasal DCR except in the case of presaccal stenosis. There are no established indications or treatment regimens for mitomycin C to date. Postoperative care after endonasal DCR should consist in the removal of fibrin, crusts and granulations and the administration of eye drops (antibiotic + cortisone) and nasal steroids. DISCUSSION: With appropriate operative technique and in experienced hands, the success rates of endonasal DCR are practically equal to those of the classical external approach. Major advantages of the endonasal approach are shorter operation times, lower complication rates and reduced patient morbidity. Neither silicone stenting nor the application of mitomycin C are routinely indicated. Laser-assisted techniques do not currently appear to improve results. Appropriate postoperative care is essential to prevent endonasal synechiae and subsequent recurrences. PMID- 14740306 TI - [Collision tumor of a hypopharyngeal adenoidcystic carcinoma and a laryngeal squamous cell carcinoma]. AB - INTRODUCTION: Malignancies in head and neck cancer are mainly squamous cell carcinomas. Adenoid cystic carcinomas are rare lesions of this site. Laryngeal adenoid cystic carcinoma is estimated to occur in 0.1 - 0.7 % of all laryngeal carcinomas. Adenoid cystic carcinomas are rarely located in the hypopharynx. To our knowledge there is no case report of adenoid cystic carcinoma of the hypopharynx as part of a collision tumor of the larynx. CASE REPORT: A 47-year old male patient was diagnosed with an adenoid cystic carcinoma of the hypopharynx and a squamous cell carcinoma of the larynx. Because of local extension of both tumors laryngectomy and partial pharyngotomy with bilateral neck dissection was performed followed by radiation therapy. Clinical aspects as well as histomorphological and immunohistochemical criteria of both tumor entities are discussed. DISCUSSION AND CONCLUSIONS: Immunohistochemical characteristics showed two different carcinoma entities in the larynx and hypopharynx. Only by complete histological investigation of a carcinoma those rare cases of a collision tumor can be detected. Both tumor entities need to be considered for therapy strategy and oncological follow-up planning. PMID- 14740308 TI - [To our Readers]. PMID- 14740307 TI - [Biallelic inactivation of the p16-Gen in a metachronous triple carcinoma in the oropharyngeal region]. AB - AIMS: Recent studies have shown that most Dutch families with atypical multiple mole melanoma (FAMMM) have a 19-bp deletion (p16-Leiden) in exon 2 of the p16 gene. Apart from reports on metachronous pancreatic tumors, other cancer types have never been described in such families. Due to heterozygous p16-Leiden constitution, our proband with multiple head and neck carcinomas was a suitable model for studying the type of p16 inactivation according to the Knudson-two-hit model. METHODS: p16 mutations in exons 1 and 2 were determined using PCR-SSCP Sequencing analysis. p16 methylation was assessed by methylation-specific PCR. RESULTS: All three metachronous (larynx, pharynx, oral cavity) tumors had a methylated p16 promotor. The p16 protein loss detected by immunohistochemistry clearly confirmed a complete loss of p16 tumor suppressor function. Thus, all three tumors exhibited biallelic inactivation of p16, caused by aberrant methylation of the p16 promotor. CONCLUSIONS: This is the first report on p16 Leiden mutation in head and neck cancer. We provide evidence that the somatic methylation of p16 promotor is associated with the germline transmission of p16 Leiden mutation. This is an example for the rare event of in which aberrant methylation acting as the 'second hit' in a familial cancer syndrome. Our results show that this epigenetic event is equivalent to genetic alterations (mutation/LOH) confirming the Knudson's hypothesis for tumor suppressor gene inactivation. PMID- 14740309 TI - [Treatment of anaphylactic reactions: a review of guidelines and recommendations]. AB - In the present review, guidelines and recommendations for the treatment of anaphylactic reactions are presented and evaluated. Herefore, publications of German and non-German societies and expert groups have been selected and, in addition to that, we have chosen a number of articles that were cited frequently and, as far as we are concerned, have a relevant meaning for this theme. None of the traditionally applied remedies - epinephrine and intravascular volume, histamine receptor blockade, inhaled betamimetics and steroids - have been proven efficacious by means of evidence-based medicine. Due to the variability of the symptoms of anaphylaxis, it is widely accepted that such a clinical proof is unlikely to come. In contrast to this, there is lack of a consense as to substantial elements of the therapy, above all with regard to dosage and application mode for epinephrine and likewise concerning amount and kind of intravenous fluids. Hence, the physician in charge has to face the necessity either to follow the guidelines of his institution (if there are any) or to blend his own therapeutic regime according to his own convictions. PMID- 14740310 TI - [Propofol-LCT versus propofol-MCT/LCT with or without lidocaine - a comparison on pain on injection]. AB - OBJECTIVE: A disadvantage of Propofol (commercial preparation containing long chain triglycerides; Propofol-LCT) is pain on injection, which is reported by 54 100 % of the patients. Many different approaches have been used in an attempt to decrease the pain on injection of propofol. The currently most efficacious treatment is lidocaine given intravenously with a tourniquet prior to Propofol injection. In previous studies, larger concentrations of free propofol in the aqueous phase of an emulsion were associated with more pain on injection. A new formulation of propofol (a mixture of long-chain and medium-chain triglycerides in the carrier emulsion; Propofol-MCT/LCT) reduces the incidence of pain on bolus injection. This study was designed to investigate, whether the use of Propofol MCT/LCT alleviates pain on injection to a similar degree as pretreatment with lidocaine. METHODS: Eighty patients (ASA I - III) were randomly assigned to four groups according to a double-blinded protocol, to receive either lidocaine 2 % or normal saline given iv. with a 60 seconds tourniquet time before the injection of Propofol-LCT or Propofol-MCT/LCT. (group 1: 2 ml NaCl 0,9 %, Propofol-LCT; group 2: 2 ml NaCl 0,9 %, Propofol-MCT/LCT; group 3: 2 ml lidocaine 2 %, Propofol-LCT; group 4: 2 ml lidocaine 2 %, Propofol-MCT/LCT). Assessment of pain on injection was performed after 30 % of the induction dose was given. RESULTS: Pain on injection caused by Propofol-LCT with pre-treatment of lidocaine and Propofol MCT/LCT alone is shown to be equivalent. Comparison of Propofol-MCT/LCT with pre treatment of lidocaine and Propofol-LCT alone shows a statistical noticeable p value of 0.035. Propofol-MCT/LCT with pre-treatment of lidocaine suggests a tendency of causing less pain compared to Propofol-MCT/LCT. Analysis of the postoperative questionnaire supplies no significant difference. CONCLUSION: The results suggest that pain on injection is reduced equivalent using either Propofol-MCT/LCT alone or Propofol-LCT with pre-treatment of lidocaine. Pre treatment with lidocaine before Propofol-MCT/LCT seems to have an additional effect. PMID- 14740311 TI - [Microbiological isolates associated with nosocomial infections in intensive care units: data of 274 intensive care units participating in the German Nosocomial Infections Surveillance System (KISS)]. AB - OBJECTIVE: To describe the frequency and distribution of microbiological isolates in association with nosocomial infections (NI) in intensive care units (ICUs) in Germany. METHODS: The hospital infection surveillance system (KISS = Krankenhaus Infektions-Surveillance-System) started in Germany in January 1997 as a nationwide surveillance project for the voluntary registration of nosocomial infections. For ICUs reporting focuses on device-associated primary bloodstream infections, urinary tract infections, as well as on pneumonia and bronchitis. The data collected for each patient who acquires a nosocomial infection include the site of infection and patient demographics. Up to 4 pathogens can be recorded for each infection. RESULTS: From 1997 until 2002 274 ICUs were participating. The data of 590 695 patients with 18 073 device associated infections (ventilator associated pneumonia and bronchitis, central line-associated bloodstream infection (BSI) and urinary catheter-associated urinary tract infection (UTI)) were analyzed. In the group of device-associated NI S. aureus was most frequently isolated (16.5/100NI) followed by Ps. aeruginosa (14.2/100NI), E. coli (13.9/100NI), enterococci (13.4/100NI) and C. albicans (11.2/100NI). Of all S. aureus isolates 20.0 % were MRSA. There was an increase of the MRSA rate from 8 % in 1997 to 26.9 % in 2002. The most frequent pathogen in BSI was coagulase negative staphylococcus (31.0/100NI). In ventilator-associated pneumonia and bronchitis S. aureus was the most frequent pathogen (24.2 and 24.9/100NI). In more than half of all catheter-associated UTI E. coli (25.7/100NI) and enterococci (25.4/100NI) were found. After stratification for the type of ICU differences in the distribution of pathogens could be shown. CONCLUSIONS: The KISS data give an overview of the most important pathogens associated with nosocomial infections on ICUs in Germany. PMID- 14740312 TI - [Prehospital emergency setting caused by hemorrhagic oropharyngeal bullae in a child with dystrophic epidermolysis bullosa Hallopeau-Siemens]. AB - Two painful large hemorrhagic bullae developed within minutes of each other in a four-month-old infant with autosomal recessive inherited epidermolysis bullosa dystrophica Hallopeau-Siemens. There was no significant impairment of spontaneous respiration. The emergency physician sedated the infant with midazolam administered by the nasal route. A community pediatrician called for assistance opened a bulla, which impressively improved the symptoms and avoided the necessity of an admission to hospital. The presented case demonstrates the importance of exchange of information within the group of emergency physicians regarding rare diseases not only in rural areas. In our opinion its also helpful to involve colleagues in private practice into the emergency system. PMID- 14740313 TI - [Airbag associated deceleration trauma with complete infradiaphragmatic dissection of the inferior vena cava (IVC) and tear of liver veins]. AB - Numerous investigations have demonstrated that the ability of passenger airbag systems to reduce both injury severity and mortality in car accidents is restricted to the simultaneous application of seatbelts and airbags. Depending on the impact force during the accident and also on the driver's position to the airbag an isolated airbag deployment can even produce severe injuries. We describe a complete infradiaphragmatic dissection of the IVC and a tear of liver veins after an airbag deployment without seatbelt use. We discuss different airbag associated injuries, the possible pathomechanism of the described injury, frequency and prognosis of abdominal vascular injuries and their emergency treatment. In the presented case the isolated severe vascular injury resulted in an intraoperative death caused by exsanguination. In blunt trauma with accompanying hypotension pre-arrival alarming of the ER, rapid transport to the hospital and immediate surgery are most important for the outcome. PMID- 14740314 TI - [Neuromonitoring for estimating the depth of narcosis]. PMID- 14740315 TI - Both rare and common polymorphisms contribute functional variation at CHGA, a regulator of catecholamine physiology. AB - The chromogranin/secretogranin proteins are costored and coreleased with catecholamines from secretory vesicles in chromaffin cells and noradrenergic neurons. Chromogranin A (CHGA) regulates catecholamine storage and release through intracellular (vesiculogenic) and extracellular (catecholamine release inhibitory) mechanisms. CHGA is a candidate gene for autonomic dysfunction syndromes, including intermediate phenotypes that contribute to human hypertension. Here, we show a surprising pattern of CHGA variants that alter the expression and function of this gene, both in vivo and in vitro. Functional variants include both common alleles that quantitatively alter gene expression and rare alleles that qualitatively change the encoded product to alter the signaling potency of CHGA-derived catecholamine release-inhibitory catestatin peptides. PMID- 14740316 TI - Evidence of a novel quantitative-trait locus for obesity on chromosome 4p in Mexican Americans. AB - Although several genomewide scans have identified quantitative-trait loci influencing several obesity-related traits in humans, genes influencing normal variation in obesity phenotypes have not yet been identified. We therefore performed a genome scan of body mass index (BMI) on Mexican Americans, a population prone to obesity and diabetes, using a variance-components linkage analysis to identify loci that influence BMI. We used phenotypic data from 430 individuals (26% diabetics, 59% females, mean age +/- SD = 43 +/- 17 years, mean BMI +/- SD = 30.0 +/- 6.7, mean leptin (ng/ml) +/- SD = 22.1 +/- 17.1) distributed across 27 low-income Mexican American pedigrees who participated in the San Antonio Family Diabetes Study (SAFDS) for whom a 10-15-cM map is available. In this genomewide search, after accounting for the covariate effects of age, sex, diabetes, and leptin, we identified a genetic region exhibiting the most highly significant evidence for linkage (LOD 4.5) with BMI on chromosome 4p (4p15.1) at 42 cM, near marker D4S2912. This linkage result has been confirmed in an independent linkage study of severe obesity in Utah pedigrees. Two strong positional candidates, the human peroxisome proliferator-activated receptor gamma coactivator 1 (PPARGC1) and cholecystokinin A receptor (CCKAR) with major roles in the development of obesity, are located in this region. In conclusion, we identified a major genetic locus influencing BMI on chromosome 4p in Mexican Americans. PMID- 14740317 TI - Pleiotropic effects of a chromosome 3 locus on speech-sound disorder and reading. AB - Speech-sound disorder (SSD) is a complex behavioral disorder characterized by speech-sound production errors associated with deficits in articulation, phonological processes, and cognitive linguistic processes. SSD is prevalent in childhood and is comorbid with disorders of language, spelling, and reading disability, or dyslexia. Previous research suggests that developmental problems in domains associated with speech and language acquisition place a child at risk for dyslexia. Recent genetic studies have identified several candidate regions for dyslexia, including one on chromosome 3 segregating in a large Finnish pedigree. To explore common genetic influences on SSD and reading, we examined linkage for several quantitative traits to markers in the pericentrometric region of chromosome 3 in 77 families ascertained through a child with SSD. The quantitative scores measured several processes underlying speech-sound production, including phonological memory, phonological representation, articulation, receptive and expressive vocabulary, and reading decoding and comprehension skills. Model-free linkage analysis was followed by identification of sib pairs with linkage and construction of core shared haplotypes. In our multipoint analyses, measures of phonological memory demonstrated the strongest linkage (marker D3S2465, P=5.6 x 10(-5), and marker D3S3716, P=6.8 x 10(-4)). Tests for single-word decoding also demonstrated linkage (real word reading: marker D3S2465, P=.004; nonsense word reading: marker D3S1595, P=.005). The minimum shared haplotype in sib pairs with similar trait values spans 4.9 cM and is bounded by markers D3S3049 and D3S3045. Our results suggest that domains common to SSD and dyslexia are pleiotropically influenced by a putative quantitative trait locus on chromosome 3. PMID- 14740318 TI - Null leukemia inhibitory factor receptor (LIFR) mutations in Stuve Wiedemann/Schwartz-Jampel type 2 syndrome. AB - Stuve-Wiedemann syndrome (SWS) is a severe autosomal recessive condition characterized by bowing of the long bones, with cortical thickening, flared metaphyses with coarsened trabecular pattern, camptodactyly, respiratory distress, feeding difficulties, and hyperthermic episodes responsible for early lethality. Clinical overlap with Schwartz-Jampel type 2 syndrome (SJS2) has suggested that SWS and SJS2 could be allelic disorders. Through studying a series of 19 families with SWS/SJS2, we have mapped the disease gene to chromosome 5p13.1 at locus D5S418 (Zmax=10.66 at theta =0) and have identified null mutations in the leukemia inhibitory factor receptor (LIFR or gp190 chain) gene. A total of 14 distinct mutations were identified in the 19 families. An identical frameshift insertion (653_654insT) was identified in families from the United Arab Emirates, suggesting a founder effect in that region. It is interesting that 12/14 mutations predicted premature termination of translation. Functional studies indicated that these mutations alter the stability of LIFR messenger RNA transcripts, resulting in the absence of the LIFR protein and in the impairment of the JAK/STAT3 signaling pathway in patient cells. We conclude, therefore, that SWS and SJS2 represent a single clinically and genetically homogeneous condition due to null mutations in the LIFR gene on chromosome 5p13. PMID- 14740319 TI - Matching strategies for genetic association studies in structured populations. AB - Association studies in populations that are genetically heterogeneous can yield large numbers of spurious associations if population subgroups are unequally represented among cases and controls. This problem is particularly acute for studies involving pooled genotyping of very large numbers of single-nucleotide polymorphism (SNP) markers, because most methods for analysis of association in structured populations require individual genotyping data. In this study, we present several strategies for matching case and control pools to have similar genetic compositions, based on ancestry information inferred from genotype data for approximately 300 SNPs tiled on an oligonucleotide-based genotyping array. We also discuss methods for measuring the impact of population stratification on an association study. Results for an admixed population and a phenotype strongly confounded with ancestry show that these simple matching strategies can effectively mitigate the impact of population stratification. PMID- 14740320 TI - Mutations within the MGC4607 gene cause cerebral cavernous malformations. AB - Cerebral cavernous malformations (CCM) are hamartomatous vascular malformations characterized by abnormally enlarged capillary cavities without intervening brain parenchyma. They cause seizures and focal neurological deficits due to cerebral hemorrhages. CCM loci have already been assigned to chromosomes 7q (CCM1), 7p (CCM2), and 3q (CCM3) and have been identified in 40%, 20%, and 40%, respectively, of families with CCM. Loss-of-function mutations have been identified in CCM1/KRIT1, the sole CCM gene identified to date. We report here the identification of MGC4607 as the CCM2 gene. We first reduced the size of the CCM2 interval from 22 cM to 7.5 cM by genetic linkage analysis. We then hypothesized that large deletions might be involved in the disorder, as already reported in other hamartomatous conditions, such as tuberous sclerosis or neurofibromatosis. We performed a high-density microsatellite genotyping of this 7.5-cM interval to search for putative null alleles in 30 unrelated families, and we identified, in 2 unrelated families, null alleles that were the result of deletions within a 350-kb interval flanked by markers D7S478 and D7S621. Additional microsatellite and single-nucleotide polymorphism genotyping showed that these two distinct deletions overlapped and that both of the two deleted the first exon of MGC4607, a known gene of unknown function. In both families, one of the two MGC4607 transcripts was not detected. We then identified eight additional point mutations within MGC4607 in eight of the remaining families. One of them led to the alteration of the initiation codon and five of them to a premature termination codon, including one nonsense, one frameshift, and three splice-site mutations. All these mutations cosegregated with the disease in the families and were not observed in 192 control chromosomes. MGC4607 is so far unrelated to any known gene family. Its implication in CCMs strongly suggests that it is a new player in vascular morphogenesis. PMID- 14740321 TI - Mutations in the VLGR1 gene implicate G-protein signaling in the pathogenesis of Usher syndrome type II. AB - Usher syndrome type II (USH2) is a genetically heterogeneous autosomal recessive disorder with at least three genetic subtypes (USH2A, USH2B, and USH2C) and is classified phenotypically as congenital hearing loss and progressive retinitis pigmentosa. The VLGR1 (MASS1) gene in the 5q14.3-q21.1 USH2C locus was considered a likely candidate on the basis of its protein motif structure and expressed sequence-tag representation from both cochlear and retinal subtracted libraries. Denaturing high-performance liquid chromatography and direct sequencing of polymerase-chain-reaction products amplified from 10 genetically independent patients with USH2C and 156 other patients with USH2 identified four isoform specific VLGR1 mutations (Q2301X, I2906FS, M2931FS, and T6244X) from three families with USH2C, as well as two sporadic cases. All patients with VLGR1 mutations are female, a significant deviation from random expectations. The ligand(s) for the VLGR1 protein is unknown, but on the basis of its potential extracellular and intracellular protein-protein interaction domains and its wide mRNA expression profile, it is probable that VLGR1 serves diverse cellular and signaling processes. VLGR1 mutations have been previously identified in both humans and mice and are associated with a reflex-seizure phenotype in both species. The identification of additional VLGR1 mutations to test whether a phenotype/genotype correlation exists, akin to that shown for other Usher syndrome disease genes, is warranted. PMID- 14740322 TI - PBAT: tools for family-based association studies. PMID- 14740324 TI - Intradialytic parenteral nutrition: where do we go from here? PMID- 14740325 TI - Assessment and monitoring of uremic malnutrition. AB - Assessment and monitoring of protein and energy nutritional status are essential to prevent, diagnose, and treat uremic malnutrition, a condition highly prevalent and associated with increased morbidity and mortality in patients with advanced kidney failure. Comprehensive assessments of protein and energy nutritional status can be achieved by several measurements to quantitatively and qualitatively estimate protein content in visceral and somatic body compartments, in addition to measurements of energy balance. However, uremic malnutrition is a complex metabolic disorder in which not only net nutrient intake is lower than nutrient requirements, leading to decreased tissue function and loss of body mass, but it is also associated with many comorbid conditions. Therefore, a clinically meaningful assessment of uremic malnutrition should include methods that are able to assess clinical outcome, identify the underlying diseases, and determine whether there is potential of benefit from nutritional interventions. Such assessment usually requires using multiple measurements concomitantly, with no definitive single method that can be considered as a "gold standard." In this review, we describe the various types of methods to assess uremic malnutrition, expanding and updating data on the readily available methods, and discuss more precise techniques to estimate protein and energy homeostasis. Special considerations of specific methods related to their clinical and/or research applicability as they pertain to renal failure are also addressed. PMID- 14740326 TI - Dietary habits and nutritional status of renal transplant patients. AB - BACKGROUND: Although dialysis nutritional problems are well described, nutritional problems after renal transplantation (RT) have received little attention. METHODS: Body composition as assessed by dual-energy x-ray absorptiometry in 115 stable patients 6.6 +/- 5.9 years after RT and repeated 2.9 years later, when a 3-day dietary history was obtained in 79 patients. RESULTS: Patients diet was generally sufficient, but was characterized by a high fat intake and deficiencies in folic acid, vitamin D, thiamine, iodine, selenium, and iron intake. Patients were often overweight, and at any given weight had a 4% to 5% higher proportion of body fat than normal. Loss of fat weight was related to high initial fat weight, long RT duration, and low plasma bicarbonate, but not steroid dose. CONCLUSION: Dietary advice concerning fat intake is indicated for RT patients, and nutritional supplements with folic acid and vitamin D are generally required. Their main nutritional problem is obesity. This is not adequately measured by body mass index, which should be supplemented by dual energy x-ray absorptiometry. Attention should be paid to the prevention of acidosis. PMID- 14740327 TI - Is subjective global assessment a reliable measure of nutritional status in hemodialysis? AB - OBJECTIVE: Subjective global assessment (SGA) is recommended in US and European guidelines for the nutritional assessment of patients with end-stage renal failure (ESRF). SGA identifies patient groups with abnormal nutritional parameters, but may fail to identify patients with malnutrition as identified by other techniques, such as total body nitrogen. We sought to compare SGA with a composite nutritional score. METHODS: HD patients were assessed by SGA, anthropometry, 3-day food diary, serum albumin, Kt/V urea, and normalized protein catabolic rate (nPCR). A composite nutritional score was derived from SGA, body mass index, percent of reference weight, triceps skinfold, midarm muscle circumference, and serum albumin. RESULTS: In 72 HD patients an abnormal SGA identified a patient group with reduced midarm circumference, midarm muscle circumference and serum creatinine and an increased composite nutritional score. However, overlap of nutritional scores was considerable between the normal and abnormal SGA groups, suggesting that SGA misclassified a large number of subjects. Serum albumin correlated with C-reactive protein (r = -0.473, P <.0001), not nutritional status. The composite nutritional score correlated with all of its components except for serum albumin. CONCLUSIONS: SGA may not reliably identify hemodialysis patients with abnormal nutrition. Serum albumin is related to inflammation and not to nutrition status. PMID- 14740328 TI - Effect of a soy protein diet on serum lipids of renal transplant patients. AB - OBJECTIVE: The aim of the present study was to evaluate the effect of a soy protein diet on plasma lipid levels of renal transplant recipients with moderate hypercholesterolemia. DESIGN: Dietary intervention case-control observational study. SETTING: Renal transplantation outpatient clinic. PATIENTS: Fifteen stable patients who had renal transplantation (serum creatinine < 2 mg/dL) with moderate hypercholesterolemia (low-density lipoprotein [LDL] cholesterol > 140 mg/dL). INTERVENTION: After a baseline dietary interview, dietary counseling was given individually with the goal of substituting 25 g of animal protein with 25 g of soy protein for a 5-week period, using commercially available soy foods, according to each patient's own preference. Main outcome measures Before and after the soy-diet period, plasma lipid profiles including total, LDL, and high density lipoprotein (HDL) cholesterol, triglycerides, apolipoprotein A1 and B were determined. Protein catabolic rate was assumed as a measure of dietary protein intake. RESULTS: Two patients dropped out. After the soy diet, total cholesterol (254 +/- 22 to 231 +/- 31 mg/dL, P <.05) and LDL cholesterol (165 +/- 20 versus 143 +/- 20 mg/dL, P <.01) decreased significantly. No significant changes were observed regarding HDL cholesterol and triglycerides. Dietary protein intake did not differ at baseline (73.2 +/- 22.9 g/day) and during the soy diet (72.6 +/- 15.6 g/day), when the reported actual soy protein intake resulted 26 +/- 8 g/day. CONCLUSIONS: This study shows that soy proteins given as part of the daily protein intake have beneficial effects on serum LDL cholesterol levels of renal transplant recipients with moderate hypercholesterolemia. Soy proteins could be of use in the nutritional management of renal transplant recipients. PMID- 14740329 TI - The effect of diet education on the laboratory values and knowledge of hemodialysis patients with hyperphosphatemia. AB - OBJECTIVE: The purpose of this study was to evaluate the effectiveness of 20 to 30 minutes per month of additional diet education on monthly laboratory values (phosphorus, calcium, parathyroid hormone, and calcium/phosphorus product) and knowledge of dietary phosphorus management in hemodialysis patients with hyperphosphatemia. DESIGN: A quasi-experimental design. SETTING: Three outpatient dialysis centers owned by the same corporation in 1 southern state. PATIENTS: Based on a 3-month average serum phosphorus >6.0 mg/dL, 70 patients were selected for participation; 63 dialysis patients completed the study, 32 in the experimental group and 31 in the control group. INTERVENTION: All patients completed a before-and-after knowledge test and had monthly blood samples drawn. Each month, the same registered dietitian provided the routine laboratory results review with control group. The experimental group received the routine laboratory review plus 20 to 30 minutes of additional diet education specifically targeting phosphorus. Main outcome measures Before-and-after knowledge test results and baseline and final serum calcium, phosphorus, parathyroid hormone, and calcium/phosphorus product levels. RESULTS: At baseline, there were no significant differences in any of the laboratory values, but the knowledge level of the experimental group was greater (P <.05) After 6 months, gains in knowledge were significantly higher in the intervention group, and the serum phosphorus and calcium/phosphorus product levels were significantly lower (P <.01) than in the control group. CONCLUSION: Based on this research, those patients who received extra education monthly showed positive changes, which may be beneficial in reducing hyperphosphatemia. PMID- 14740330 TI - Postdialysis albumin: a better nutrition marker in pediatric hemodialysis patients? AB - BACKGROUND: Prehemodialysis serum albumin is considered an important indicator of nutritional status in pediatric hemodialysis (HD) patients. Study aim To determine whether serum albumin levels changed significantly from predialysis to postdialysis, and correlated with fluid changes over HD treatment. METHODS: We prospectively measured pre-HD and post-HD albumin levels in 9 pediatric patients (ages 10 to 20 years; mean age, 16.3 years) on HD over an 8-month period. Changes in weight and albumin over 45 HD treatments were compared. RESULTS: A total of 66.7% of pre-HD and 100% of post-HD albumin levels were within the normal range. Post-HD albumin was significantly higher than pre-HD in 42 of 45 observations (Mean, 4.7 +/- 0.39 g/dL versus 3.8 +/- 0.37; P <.0001). Delta albumin/treatment inversely correlated with weight loss (r = 0.741, P <.0001) and prescribed ultrafiltration rate (r = 0.764, P <.0001) CONCLUSIONS: Serum albumin levels increase significantly postdialysis and inversely correlate with fluid removal. Pre-HD albumin level may not be a valid indicator of protein status because of the confounding effect of interdialytic weight gain, and may instead be a marker for fluid overload. Further studies are needed to assess the value of postdialysis albumin level as an indicator of nutritional status in pediatric HD patients. PMID- 14740331 TI - Patient education. Incentive program to control interdialytic weight gains. PMID- 14740342 TI - [Erectile dysfunction -- still a taboo subject]. PMID- 14740343 TI - Genetic factors in assisted reproduction. AB - It is still unclear whether the procedures of assisted reproduction increase the risk of congenital malformations. Thus, it remains to be clarified whether an increased risk, if any, of congenital malformations in these children is caused by the procedure of assisted reproduction itself or by the underlying maternal and paternal background. From the genetic point of view, infertility patients seeking assisted reproduction have to be classified as a high-risk group. The prevalence of numerical chromosomal abnormalities is around 10% in these patients, compared with 0.85% in the general population. The prevalence of structural chromosomal abnormalities is around 0.1% in the general population and is increased up to 1% in patients seeking assisted reproduction. In addition, patients with microdeletions of the Y-chromosome or mutations in the cystic fibrosis transmembrane-conductance regulator gene are likely to be encountered at the fertility clinic. Therefore, genetic screening and counselling should be routinely offered to infertility patients. They also need to understand that parental factors can be transferred to offspring that would most likely not have been conceived by natural means. PMID- 14740345 TI - Prevalence and risk factors for erectile dysfunction in Austria--analysis of a health screening project. AB - PURPOSE: The aim of this study was to determine prevalence and risk factors for erectile dysfunction (ED) in men undergoing a health investigation. METHODS: Men aged 30-69 yrs participating in a health screening project in the area of Vienna completed a 11-item questionnaire on ED which was extracted from the international index of erectile function (IIEF). In parallel, all men underwent a detailed health examination including physical assessment, evaluation of various life-style factors and a blood analysis with 14 parameters. RESULTS: Within the total study population (n = 832; 45.3 +/- 9.8 yrs), 8.2% reported no sexual intercourse during the previous four weeks, 5.1% reported moderate/severe ED, 13.1% mild and 73.6% no ED. ED, defined as any problem in maintaining erection until the end of sexual intercourse, increased from 12.5% in those aged 30-39 yrs to 15.3% at 40-49 yrs, 27.4% at 50-59 yrs and 45.2% in men 60-69 yrs. In parallel, the percentage of men without sexual intercourse during the previous four weeks increased from 7.4% at 30-39 yrs to 8.8% at 50-59 yrs and 17.0% at 60 69 yrs. By far the strongest correlate for ED was age; other factors were cardiovascular diseases, physical activity and diabetes mellitus. CONCLUSIONS: This study provides the first estimate of the prevalence of ED in different life decades of men in Vienna, and identifies a number of risk factors for this highly prevalent disorder in ageing men. PMID- 14740344 TI - [Insulin resistance in polycystic ovary syndrome]. AB - With a prevalence of 5-10% the polycystic ovary syndrome (PCOS) is a common disorder of premenopausal women. According to prospective studies abnormal glucose tolerance and diabetes mellitus present in about 10.0% and 35.0% of adult women with PCOS, respectively. PCOS patients have a higher prevalence of cardiovascular risk factors such as hypertension and dyslipidaemia. The rate of spontaneous abortions as well as the risk of developing gestational diabetes is increased in PCOS. Therefore, PCOS is not only a reproductive problem, but a complex endocrine disease with important health implications. The role of glucose metabolism in PCOS, the health consequences and possible interventions are reviewed in this article. PMID- 14740346 TI - Absence of microdeletions in the azoospermia-factor region of the Y-chromosome in Viennese men seeking assisted reproduction. AB - PURPOSE: The azoospermia-factor region of the Y-chromosome is essential for spermatogenesis in humans. In the literature, a wide range is given for the frequency of microdeletions in this region. The purpose of this study was to evaluate our own population of patients. METHODS: During a two-year period at Vienna Medical School, all male patients (n = 383) seeking assisted reproduction were screened for microdeletions. Thirty-three men had azoospermia and 154 severe oligozoospermia. Genomic DNA was prepared from peripheral lymphocytes and polymerase chain reaction analysis of the azoospermia-factor region was performed using the Promega kit. RESULTS: No case tested positive for azoospermia-factor microdeletions. In all cases amplification of 18 non-polymorphic sequence tagged sites was obtained. CONCLUSIONS: Y-chromosome microdeletions do not seem to be an important factor for male infertility in our patients. This suggests that screening should be restricted to men with azoospermia or severe oligozoospermia only. PMID- 14740347 TI - Angiotensin-converting enzyme genotype, albuminuria and plasma fibrinogen in type 2 diabetes mellitus. AB - AIMS: Increased fibrinogen level is considered an important atherosclerosis risk factor. Patients with type 2 diabetes frequently have increased fibrinogen levels. The aim of the present study was to examine the effect of angiotensin converting enzyme (ACE) gene polymorphism and the effects of the diabetic environment on plasma fibrinogen in type 2 diabetes. PATIENTS AND METHODS: The study group included 125 patients with type 2 diabetes (40 men, 85 women). The average age of patients was 62 +/- 10 years. Fibrinogen concentration was determined with the thrombin coagulation test. ACE insertion/deletion (I/D) polymorphism was detected using polymerase chain reaction (PCR) assay. RESULTS: II homozygotes (n = 17) had the highest mean fibrinogen levels, ID heterozygotes (n = 75) had medium levels and DD homozygotes (n = 33) had the lowest (p = 0.054, ANOVA). II homozygotes also had significantly higher mean fibrinogen level than ID/DD carriers (4.3 +/- 1.7 vs. 3.5 +/- 1.3 g/l; p = 0.015). The indices of renal functions, i.e. albuminuria (r = 0.37; p < 0.0001) and serum creatinine (r = 0.22; p = 0.015), significantly correlated with fibrinogen levels. The correlation between albuminuria and fibrinogen was significant in the subgroups with genotypes II (r = 0.76; p = 0.001) and ID (r = 0.37, p = 0.002), whereas in the subgroup of DD homozygotes this relationship did not reach statistical significance. In the multivariate regression analysis with age, sex, BMI, creatinine, albuminuria and ACE genotype as independent variables, albuminuria was the only significant predictor of fibrinogen level (p < 0.0001). After interaction between the ACE genotype and albuminuria was included into multivariate analysis, the interaction became the only independent predictor of plasma fibrinogen level (p < 0.0001) in the model, and the model explained 25% of the plasma fibrinogen variance. CONCLUSION: ACE gene polymorphism is associated with plasma fibrinogen level in type 2 diabetes. This association is mediated by an interaction between ACE genotype and albuminuria. Diabetes patients with genotypes II or ID have increased plasma fibrinogen in the presence of albuminuria. PMID- 14740349 TI - Overcoming the downside of asymmetry. PMID- 14740348 TI - [Endoscopic palliation of malignant gastric outlet obstruction by self-expanding metal stents]. AB - INTRODUCTION: Malignant gastric outlet stenosis is caused by tumour obstruction and restricts the oral intake of food, resulting in a seriously reduced quality of life. Endoscopic implantation of self expanding metal stents (SEMS) can clear stenosis in the GI-tract and reestablish and preserve the passage in the GI tract. PATIENTS AND METHODS: Between October 2001 and April 2003 seven patients with malignant gastric outlet stenosis have been treated by the implantation of SEMS. Four patients had malignant stenosis in the upper duodenum or gastric antrum, two patients had stenosis because of tumour recurrence in the efferent loop of the jejunum after gastric resection because of gastric carcinoma and one patient had an obstruction 20 cm distal of the oesophagus after gastrectomy because of gastric cancer. RESULTS: In all patients obstruction was cleared by the implantation of SEMS, and oral intake of food was possible in all patients after two days. No serious complications occurred during or after stent implantation. CONCLUSION: Stent implantation for the treatment of malignant gastric outlet stenosis is a cost effective procedure, associated with low risk and low stress for the patient, and provides excellent palliation of symptoms in patients with malignant gastric outlet stenosis. PMID- 14740350 TI - Correlates of children's competence to make healthcare decisions. PMID- 14740351 TI - Making wishes known: the role of acquired speech and language disorders in clinical ethics. PMID- 14740352 TI - Opportunities for advance directives to influence acute medical care. PMID- 14740353 TI - Exploring asymmetry in the relationship between patients and physicians. PMID- 14740354 TI - A bridge to nowhere. PMID- 14740355 TI - Mrs. T's story: an interview. Interview by Rebecca Horr, Lauren Kattany, Ellen M. Robinson. PMID- 14740356 TI - The nurses' story about Mr. T. PMID- 14740357 TI - Ethical issues raised by LVADs and Mr. T's story. PMID- 14740358 TI - Hope or truth: commentary on the case of Mr. T. PMID- 14740359 TI - Introduction. Auxology: spanning mechanism and measurement. PMID- 14740360 TI - Prenatal effects of intra-uterine growth retardation on adult height of conscripts from Hungary. AB - Physical development appropriate for age, and the normal rate of development of children, are two well-known indicators of the biological status of populations. Physical development of children is influenced by several factors, and the intrauterine environment may be critical among them. The authors studied the effect of the prenatal environment, as measured by birth length and birth weight, and socio-economic factors, as measured by place of residence and educational level of parents, on the height of 18-year-old conscripts surveyed in 1998. The following results were obtained: 1. The conscripts were classified into one of the following groups: small for gestational age (SGA), appropriate for gestational age (AGA), large for gestational age (LGA) according to their data of birth and their development at birth. Statistically significant differences in birth length and height at 18 years of age were found for the AGA and LGA groups, according to place of residence at birth. In addition, nutrition at birth (and the biological and social inclusion related to this) affects the extent of change in physique between birth and 18 years of age, within each group. The extent of change in height in the SGA group is 122.38 cm, 123.40 cm in the AGA group, and 124.11 cm in the LGA group. With each neonatal developmental group, conscripts from Budapest had the highest values both in body length and height at 18 years of age. 2. The educational level of parents influenced the physical development of their children. Means of birth length, and of body height at the age of 18 years, were greater, the higher the level of education of their parents. The lower the level of education the parents have, the more significant is the difference between birth length and height at the age of 18 years, compared to the sample mean. This is attributed to a health-cultural-information deficit arising from the low level of education of the parents. PMID- 14740361 TI - The regulation of bone development as a biological system. AB - A large number of molecular, cellular, and epidemiologic factors have been implicated in the regulation of bone development. A major unsolved problem is how to integrate these disparate findings into a concept that explains the development of bone as an organ. Often, events at the organ level are simply presented as the cumulative effect of all factors that individually are known to influence bone development. In such a cumulative model it must be assumed that each bone cell carries the construction plan of the entire skeletal anatomy in its genes. This scenario is implausible, because it would require an astronomical amount of positional information. We therefore propose a functional model of bone development, which is based on Frost's mechanostat theory. In this model, the genome only provides positional information for the basic outline of the skeleton as a cartilaginous template. Thereafter, bone cell action is coordinated by the mechanical requirements of the bone. PMID- 14740362 TI - Quantitative and molecular genetic determination of protein and fat deposition. AB - After 30 years of selection, breeding of the pig breed sus scrofa Pietrain has resulted in reduced backfat thickness (from 3.2 to 1.9 mm) and increased loin muscle area (40 to 60 cm2) which indicates high genetic determination of these body composition traits. The use of sophisticated quantitative genetic methods that include all genetic relationships of large populations has led to a high response to selection of these traits. Selection on feed intake, lean and fat tissue growth using nonlinear functions to optimise these traits during the entire growth period in a biological model offers the opportunity to further improve total genetic potential. Protein and lipid deposition rates during the entire growth period have to be known for this biological model to be applied; thus knowledge of the genetic background of these traits is of high economic value. With the use of molecular genetic methods, such as candidate gene and genome scan approaches, the identification of genes for obesity and growth can be obtained. In sus scrofa, candidate genes associated with obesity and growth include Leptin Receptor, Melanocortin-4 Receptor, Agouti related protein, Heart fatty acid binding protein 3, and Insulin-like growth factor 2. Some of these candidate genes also explain variation in obesity levels in humans. Initial genome-wide scans have identified quantitative trait loci (QTL) on chromosomes 1, 4, 5, 7 and X for obesity and on chromosomes 1, 4, 7, 8, 13 and 18 for growth. Physiological candidate genes and predispositional QTL for obesity are not always located on the same chromosome; this is known the "polygenic paradox". Use of a nonlinear growth function is recommended in order to give more insight into the physiological regulation of obesity traits. Sus scrofa is an excellent model organism to examine the genetic regulation of obesity. The conservation of DNA sequence and chromosomal segments between sus scrofa and homo sapiens will permit easy transfer of results to human studies. PMID- 14740363 TI - Growth hormone and aging. AB - In elderly people, vascular alterations and degenerative alterations of the Central Nervous System (CNS) are two of the most common reasons for illness and death. Lipid pattern modifications and menopause in women are some of the causes for the appearance of these alterations. Vascular endothelium is in part responsible for vascular homeostasis, through the production of several vasoactive factors. Growth hormone (GH) exerts effects on the CNS and on the vascular endothelium, since GH deficient subjects exhibit endothelium-dependent alterations, which recover under substitutive GH treatment. Growth hormone has important actions on lipid metabolism that also play a role on vascular and endothelial function. Moreover, cardiac function improves when GH is associated to angiotensin II receptor blockers. Elderly people exhibit a physiological GH deficiency that could affect their vascular and cerebral functions. A study was carried out using old Wistar rats to clarify the effects of GH on the vessels under chronic "in vivo" conditions. The response to various vasoactive substances in aortic rings has been evaluated. An increase in the aortic media thickness was seen in old rats, which showed also a reduction in the vasodilator response to isoprenaline as compared to young animals. GH treatment partially restored the vasodilator response and reduced media thickness. Neuronal population was reduced in the hypocampus of old rats as compared to young ones and GH treatment was able to significantly enhance the number. Neurotransmitters were measured in several cerebral areas to establish differences between young and old GH-treated or untreated animals. Glutamine, Arginine and Aspartate were reduced in old animals whereas Citruline was increased. GH treatment restored in all cases the levels corresponding to young rats. PMID- 14740364 TI - Synthetic standards for body weight. AB - Growth charts represent body stature, body weight, and body mass index (BMI) from birth to maturity. Due to secular changes in these parameters, growth charts tend to become outdated, and must be revised from time to time. Recently, we developed alternative strategies that facilitate developing and renewing growth charts, and suggested synthetic standards for body stature. The increasing prevalence of obesity has made it necessary to develop similar techniques also for monitoring body weight and BMI. Two-hundred-and-forty historic and modern growth studies (108 studies of male growth, 132 studies of female growth) were selected from 22 European, 6 American, 3 African, and 6 Asian nations, published between 1831 and 2001. The studies contained annual information on weight and stature, either between birth and 6 years, or between 6 years and maturity, or information on the whole age range between birth and maturity. Since historic studies up to the mid 20th century usually ignore the fact that body weight (in contrast to body stature) is not normally distributed, a group of 92 more recent studies (45 male, 47 female), published between 1943 and 2001, presenting centiles for weight, was chosen for additional analysis. Furthermore, the skewness of body weight distributions, was investigated in original raw data of body weight obtained from five well reputed longitudinal growth studies, performed at Jena, Germany, Lublin, Poland, Paris, France, Prague, Czech Republic, and Zurich, Switzerland. Average body stature and average weight differ markedly between different populations. But within the same population, both parameters are closely interrelated. In males, birth length and weight correlated with r = 0.503, stature and weight correlated with r = 0.873 at the age of 2 years; with r = 0.882 at the age of 6 years; with r = 0.935 at the age of 14 years, and with r = 0.891 at the age of 18 years. Similar results were obtained in females. At birth, length and weight correlated with r = 0.619. Stature and weight correlated with r = 0.863 at the age of 2; with r = 0.912 at the age of 6; with r = 0.935 at the age of 12; and with r = 0.918 at the age of 15 years. Tables of linear regression coefficients for relative stature and weight at all ages enable the reversal of the process of the meta-analysis and allow the generation of synthetic growth references for stature and weight. Synthetic reference charts help in the revision of current growth charts without much additional effort, and may be used for populations for which autochthonous growth standards are not available. PMID- 14740365 TI - Automatic analysis of longitudinal growth data on the Website willi-will wachsen.de. AB - Many disorders of child development can only be treated successfully when they are detected early. Thus, child development should be checked periodically. Usually, a few parameters are sufficient to check whether or not a child is developing normally in terms of growth. By making such checks publicly available on the website: willi-will-wachsen.de the authors hope to provide a tool which helps the automatization of simple check procedures and thereby detect with less effort more children with growth disorders. PMID- 14740366 TI - Forensic age diagnostics of living individuals in criminal proceedings. AB - Age estimations of living individuals are increasingly important in criminal matters. If doubts arise regarding the age of a person suspected of a criminal offense, forensic age estimation is prompted by the need to ascertain whether the person concerned has reached the age of criminal responsibility and whether general criminal law in force for older juveniles or adults is to be applied. According to the recommendations of the Arbeitsgemeinschaft fur Forensische Altersdiagnostik (study group for forensic age diagnostics), age estimates in criminal proceedings should be based on the general physical examination, the X ray examination of the hand and the odontological examination by a dentist, including dental status and orthopantomogram. In order to improve diagnostic reliability, these methods should always be used in combination, ensuring that each part is performed by forensically trained and experienced experts of the relevant disciplines. In order to demonstrate that the proband has reached the age of 21, an additional X-ray examination or CT scan of the clavicles is recommended. Future research projects should assess the variation for a combination of the above methods, quantify the impact of socio-economic status and ethnicity on the examined development systems and review the suitability of non-ionizing imaging methods of age estimation. PMID- 14740367 TI - Nutritional status in two Mbya-Guarani communities from misiones (Argentina). AB - Growth and nutritional status of children and adults in two Mbya-Guarani communities from Argentina, was assessed. Height, weight, sitting height, upper arm circumference, triceps and subscapular skinfold were measured on 120 individuals aged from 2 to 60 between March and May, 2001. Data were transformed to z-scores using United States references (NHANES I and NHANES II). A z-score of less than -2 was used as the cut-off point to determine the prevalence of stunting and wasting respectively. Mean z-scores for weight, height, and upper arm circumference lie below the reference (0 > Z > -3), while in sitting height ratio and muscular area for females they were above the US standard (0 < Z < 2). Skinfold thicknesses and muscular area for males were similar to the reference (2 > Z > -1). Our findings are in agreement with others South American Indian research that the prevalence of stunting (36.7%) is significantly higher than wasting (1.8%). The presence of parasitic infections and nutritional environment previously described in this population could be related to the short stature. PMID- 14740368 TI - [Mainstreaming of young handicapped students. Mainstreaming in the ordinary milieu should be privileged]. PMID- 14740373 TI - [Legal framework for adoption]. PMID- 14740374 TI - [Need for agreement, a balance between evaluation and caring]. PMID- 14740375 TI - [The adoptive family in crisis, family therapy, an adaptive response]. PMID- 14740376 TI - [Adoption by homosexual parents]. PMID- 14740377 TI - [Adopting is to embrace]. PMID- 14740378 TI - [A partial observation of an original experience]. PMID- 14740379 TI - [Management of a crisis situation in school, systematic approach]. PMID- 14740380 TI - [6/7 Posture during perfusion]. PMID- 14740381 TI - [Differed emergencies. 2/ Hirschsprung's disease]. PMID- 14740382 TI - [Importance of diagnostic and therapeutic pars plana vitrectomy in endogenous uveitis]. AB - OBJECTIVE: To evaluate the importance of diagnostic-therapeutic pars plana vitrectomy in patients with clinical manifestations of endogenous uveitis. PATIENTS AND METHODS: The diagnostic-therapeutic pars plana vitrectomy (DT PPV) has been performed at the Eye Clinic of General Faculty Hospital and 1st Medical Faculty on Prague on 62 eyes in 49 patients at the mean age of 45 years. The DT PPV was indicated in patients with uveitis of unusual clinical picture or poorly reacting to treatment, suspected infection or tumor etiology. In the beginning of PPV, 0.5 to 1.5 ml of undiluted vitreous body was samples and subsequently examined by microbiology, immunology and molecular genetic methods. PPV was performed on both eyes in 10 patients, on one eye twice in each of three patients. The authors evaluated 62 samples of vitreous body on the whole. The patients were observed for up to 96 months. RESULTS: Laboratory examination of the samples of vitreous body from 22 eyes (35.4%) supported our notion on infection etiology of intraocular inflammation, manifestations of intraocular tumor were in 12 eyes (19.3%), vascular or vitreoretinal pathology was present in 6 eyes (9.6%). Nonspecific inflammatory changes were detected in 22 eyes (35.4%). Drugs were administered into the vitreous body in 16 eyes. In the observation period the authors detected marked late complications in connection with DD PPV in 18 eyes (29%). Visual acuity improved by two or more lines of the Snellen optotypes in 43 eyes (69.3%), remained unchanged in 11 (17.7%) eyes. The vision deteriorated in 8 eyes (12.9%). CONCLUSION: Based on this experience the authors are of the opinion that DT PPV is indicated in patients with uveitis for giving more precision to the diagnosis of intraocular inflammation or a masking syndrome. DT PPV also represents a therapeutic procedure to cure complications connected with a severe form of inflammation and makes it possible to save central visual acuity. PMID- 14740383 TI - [Experimental correction of irregular astigmatism in patients with keratoconus using diode laser thermal keratoplasty]. AB - OBJECTIVE: To evaluate the potential of diode laser thermokeratoplasty (DTK) in the correction of irregular astigmatism in patients with keratoconus. GROUP AND METHODS: The coagulation of deep layers of cornea stroma was made on four eyes of four patients suffering from advanced keratoconus at the mean age of 31 years by means of the infrared contact cw laser Prolaser Rodenstock 1.9 DTK. The aim of the treatment was to increase the curvature of cornea in flat meridians and to compensate flattening of cornea in steep meridians by way of forming traction strips between the contracted tissue of individual coagulation points. The changes on the cornea topography and changes of visual functions were determined. RESULTS: The diode laser keratoplasty is capable to induce sector increase of cornea curvature and thereby improve symmetry pictures of irregular astigmatism in keratoconus. For a definitive inclusion of DLK into the spectrum of clinically applicable procedures, other therapeutic studies are needed as well as larger groups with relatively homogeneous pre-operation pictures. PMID- 14740384 TI - [Stability of corneal optic power after photorefractive keratectomy]. AB - Changes in the values of optical power of cornea in patients after photorefractive keratectomy (PRK) obtained by Fourier analysis of videotopographic picture of cornea were investigated. The authors compared changes of cornea surface (optical power) in patients with preoperation myopia up to 6 and more than 6 diopters. Moreover, the changes of absolute values of corneal optical power were compared with the development of refraction of the optical system as a whole. In patients with a lower degree of myopia the cornea optical power became stable up to 6 months after the operation. The cornea surface in patients with higher myopia was still changing one year after the operation. The changes of optical power of cornea in patients with a higher myopia were more marked, but the differences were not statistically significant (except for the period after cornea ablation). There was not any significant difference between the changes of objective refraction and the changes of cornea optical power. The main cause of fluctuating postoperative refraction is apparently the change of cornea surface associated with healing of cornea. PMID- 14740385 TI - [Is it possible to accurately measure intraocular pressure after refractive surgery?]. AB - OBJECTIVE: To evaluate how the change of thickness or curving of the cornea after refraction operations (PRK or LASIK) influences the intraocular pressure (IOP) during assessment with a noncontact tonometer. PLACE OF INVESTIGATION: Ophthalmological Clinic SPAM and Ophthalmological ambulance EXCIMER. MATERIAL AND METHODS: With regard to the performed refraction operation the patients were divided into two groups. The first group comprised 87 patients where PRK was performed to correct myopia. The mean myopia in this group was -5.27 D +/- 2.32. The second group comprised 62 patients where LASIK surgery was performed to correct myopia. The mean myopia in this group was -10.18 +/- 4.36. IOP was assessed by means of a noncontact tonometer before and 6 months after surgery. RESULTS: In the first group a mean reduction of IOP by 4.8 +/- 2.1 mm Hg occurred. The central thickness of the cornea declined on average by 55.69 +/- 28.9 micros and the curving of the cornea diminished on average by 3.72 D +/- 1.76 D. In the second group of patients the IOP declined on average by 7.97 +/- 2.04 mm Hg. The central thickness of the cornea diminished by 78.75 microns +/- 42.26 and the curving of the cornea by 4.65 D +/- 2.07. In both groups a significant correlation was found between the reduction of IOP and the central thickness and curving of the cornea. CONCLUSION: Diminution of the thickness of the central cornea and diminution of the curving lead to lower values of the IOP. There is a direct correlation between thinning of the cornea and the decline of IOP. PMID- 14740386 TI - [Indications for refractive procedures in adult patients with strabismus and results of the subsequent therapeutic procedures]. AB - The authors estimated the contribution of the refraction intervention and a possible evolution in the position of eyes in 102 adult individuals considering a refraction intervention. The indications for this intervention were a disorder in eye position or amblyopia in those who were interested in this operation and were examined in the years 1996 to 2002. The basic refraction examination was always supplemented by a detailed orthoptic analysis. Based on this examination the intervention was not recommended in 14 examined subjects (14%). ARK represented the contraindication of the refraction intervention in 9 patients, since subsequent changes in the size of the deviation or operation adjustment of strabismus could result in diplopia. An excessive convergence with a high AC/A in hyperopia was also considered as an unsuitable indication, since a lasting cosmetic and functional significant convergent deviation into near distances could not be excluded. Five patients declined from the refraction intervention on the basis of this explanation. The paper is mainly dealing with an analysis of the development of position of the eyes and binocular functions in 46 adult patients, who decided to undergo a refraction intervention and further orthoptic care after a complex stroboscopic and refraction examination. The adjustment of refractive error was made by the LASIK methods (Laser in Situ Keratomileusis) on 69 eyes (80%) and CLE (Cleans Lens Extraction) on 17 eyes (20%). The orthoptic analyses before and the refraction intervention revealed that in all 29 even only partially accommodating esotropia, the deviation was diminished after the refraction intervention on the average by +11.2 degrees (in the rage of +2 degrees to +30 degrees) to 5.4 degrees (in the range of parallel position to +20 degrees) in the predominantly represented hyperopia, but also in 6 myopias. The improved position of the bulbs was not directly associated with the degree of hyperopia with the original deviation. The deviation after the refraction intervention in 23 patients (79%) with esotropia was not higher than degrees. The cosmetic position of the eyes was completely satisfactory and did not therefore represent even indication for the operation. The 9 patients (35%) with esotropia and hyperopia there were an improvement of binocular functions. It could be theoretically due to the newly developed emetropia making permanent optimal sensory information possible. In all 17 exotropias there were not any significant changes in the size of deviation and the binocular functions were not reestablished, if they were not retained before. The position of eyes was solved surgically in 12 patients, while exotropia predominated in two thirds of them. The weakening or strengthening interventions on horizontal straight muscles were selected according to character of strabismus in 11 patients. Recession of the lower oblique muscle was indicated one case only for the simultaneously present torticollis with exotropia. The residual deviation was not greater than 5 degrees immediately after the operation or during the following months. Binocular functions were not reestablished in any patients. A alternate suppression or suppression of perception on one eye were proved. PMID- 14740387 TI - [Hang-back retroposition as a variation in oculomotor muscle weakness]. AB - In a clinical study the authors evaluate efficiency of the operation on oculomotoric muscles by the hang-back retroposition method. A total of 70 children have been operated on, 52 of the patients suffering from esotropia and 18 from exotropia. The mean preoperation deviation in esotropia was 30 +/- 12 pdpt, the postoperation deviation was in the range of 6 +/- 7 pdpt. In the exotropia cases the preoperation deviation improved from original 76.9%, in the exotropia group being 77.8%. The overall success rate of the hang-back retroposition in all these patients was 77.1%. Ina group of children operated on by a traditional retroposition there was an overall success rate 78.7% (77.6% in esotropia and 82.4% in exotropia cases). Hang-back retroposition has been equally effective as the traditional retroposition of oculomotoric muscles. Its main advantages include visibility of the operation field, minimum risk of perforation of sclera and a fair time factor of the intervention. The authors discuss the development, indications, further advantages but also possible disadvantages or limitations of this acknowledged surgical procedure in strabology. PMID- 14740388 TI - [The pseudoexfoliative syndrome and its significance in cataract surgery]. AB - OBJECTIVE: To find out factors or combinations of factors leading to increased occurrence of complications in operation on cataract in patients with pseudoexfoliative syndrome. SETTING: Eye Clinic SPAM, Bratislava. METHODS: Prospective non-randomized study in 2916 patient of different age and sex, followed for 6 months after the operation on cataract. The standardized operation protocol was used. Complications were followed during and after the operation in view of released supporting apparatus, loss of vitreous body, uveal reaction and the occurrence of secondary cataract. RESULTS: PEX occurred in 2.98% of the surgically treated patients of the group, most often in the age group of 75-84 years. The occurrence was independent of sex, but increased with age. There was not any relation between age and the type of cataract, incidence of complications, inflammatory reaction, and width of pupil, endo-telepathy and glaucoma. The incidence of per-operation complications in 13.7% is directly related to the syndrome, a narrow pupil, disorder of supporting (suspension) apparatus and worse visual acuity. From the 34 patients with PEX and increased inflammatory reaction, 22.9% suffered from secondary cataract after 6 months. A relationship between width of the pupil, time factor, hardness of cornea and the incidence of inflammatory reaction was demonstrated in 24.1%. CONCLUSION: PEX itself is the source of most severe complications in operation on cataract. The patients should be in dispensary care and operated on in time by an experience surgeon. PMID- 14740389 TI - [Initial experience with the EAS-1000 (NIDEK) anterior segment analyzer]. AB - The analyzer of anterior segment EAS-1000 is one of recently developed diagnostic devices, used for digital photography for the analysis and storage of data about the anterior segment of the eye. The aim of the contribution is to inform the public about possible use of the apparatus in clinical and field practice. PMID- 14740390 TI - [Eye injuries in famous persons in Czech history]. PMID- 14740391 TI - Evidence-based dentistry. PMID- 14740392 TI - The single-unit crown: factors for clinical success. AB - Application of the crown preparation guidelines obtained through theoretical models, laboratory testing and clinical studies provide an evidence-based rationale for clinical practice procedures. The evidence-based rationale has been presented to support basic day-to-day techniques in clinical practice. As new techniques and materials are developed, each should be evaluated carefully prior to clinical use and acceptance. PMID- 14740393 TI - Evidence-based practice in removable prosthodontics. AB - This review of selected articles has discussed the best evidence from clinical studies pertaining to removable prosthodontics. The goal was to correlate recent literature in context with what is clinically significant for the general dentist practicing removable prosthodontics. The focus of this limited review should encourage more interest in seeking valid literature to support and incorporate evidence-based practices into daily prosthodontic treatment decisions. Prosthodontic treatment planning is a complex process that involves a combination of diagnostic information, patient desires, evidence-based outcome data, and a thorough review of the treatment alternatives (1). With appropriate skills and the availability of software to conduct a literature search, evidence-based practice is a powerful means for general dentists to establish the effectiveness of patient treatment decisions and enhance clinical knowledge and skills over the course of a professional career (5). PMID- 14740394 TI - The partially edentulous patient: fixed prosthodontics or implant treatment options. AB - The replacement of missing teeth by conventional fixed partial dentures or by implant supported prostheses occupies a major portion of the average restorative and prosthodontic practice. New materials, techniques, and technologies are constantly being added to our armamentarium. The demand for fixed prosthodontic patient services will continue to grow. It is the ultimate duty of the provider of these services to utilize an evidence-based rationale when determining the best possible treatment for the partially edentulous patient. PMID- 14740395 TI - This one's too big, this one's too small.... PMID- 14740396 TI - Oral and maxillofacial pathology case of the month. Snuff dippers keratosis or snuff pouch. PMID- 14740397 TI - Sitting up straight: why seated posture isn't what it used to be. PMID- 14740398 TI - [Safety information project on drug, food and chemicals. Division of Safety Information on Drug, Food and Chemicals. National Institute of Health Sciences]. AB - Recent issues on BSE(Bovine Spongiform Encephalopathy) and health hazards caused by adverse reactions of medical drugs, have strongly emphasized the necessity for safety measures to secure public health. These issues have been attributed to the delay to obtain overseas information on safety and regulation, and the lack of an adequate system for acquirement and assessment of such information. In order to develop a system where domestic and international safety information is collected, analyzed, assessed and presented both scientifically and systematically, the Division of Chem-Bio Informatics of the National Institute of Health Sciences was reorganized to the Division of Safety Information on Drug, Food and Chemicals in April, 2003. Collection and evaluation of safety information on medical drugs, food and chemical substances is now centralized at the Division, which consists of 5 sections, the first, second and third sections being newly established. The first section assesses information on medical drugs, the second section deals with food microorganisms, and the third section focuses on chemicals in food. The fourth and fifth sections retain their previous functions, namely, chemical safety information research and information network infrastructure support within the institute, respectively. The purpose of this paper is to describe how we will manage safety information on drug, food and chemicals, focusing on the role of the three new sections. PMID- 14740400 TI - [Examination related to revised test method for determination of formaldehyde, regulated by the law for the control of household products containing harmful substances]. AB - In Japan, the amount of formaldehyde in textile products was regulated by the low for the control of household products containing harmful substances. Formaldehyde was determined by measuring the optical density of acetylacetone derivative of formaldehyde extracted from textiles. The household products low stated that the increase in the optical density of color development of the extract from the textile products for babies or infants within 24 months after birth should not be more than 0.05. Collaborative study decided the amount of formaldehyde equivalent to the increase in absorbance described above, and the amount was 16 ppm. There are some reports that formaldehyde causes an allergic reaction even at a very low concentration, so continuous regulation for formaldehyde in the textiles was desirable using this level of amount. We developed HPLC method for the determination of formaldehyde in textile products. Formaldehyde was determined by the direct injection of acetylacetone derivative of samples into the system equipped with ODS column and UV-VIS detector (detection wavelength 413 nm) using the mixture of acetonitrile and water as mobile phase. The linearity was obtained between a peak area or height and the concentrations of formaldehyde solution in the range of 0.0625-2 micrograms/ml. The regulation level was sufficiently detected by the present HPLC method. We recommended that the HPLC test was adopted as a reexamination method for the products may violate the regulation as well as a dimedone test. PMID- 14740399 TI - [Influence of fertilizers on yield and yield components of opium from Papaver somniferum Linn]. AB - The effect of fertilizer on the yield and yield components of opium from Papaver somniferum L.cv.Ikkanshu were investigated from 1993 to 1995. The weight of opium per one capsule was tendency to increase in the order of peat moss > pulverized charcoal > compound fertilizer, but the differences among the results were not significant. Furthermore, no difference in the capsule weight per 100 m2 and the yields of opium and alkaloids per 100 m2 were detected. The significant difference was only observed when the data were compared among the weights and yields of different years. The difference among the yield of opium was mainly due to the weight of opium per one capsule. The weight of opium per one capsule showed a high correlation with capsule husk weight. The alkaloids contents in opium obtained with a different fertilizer application showed no difference. Morphine content at the first lancing was about 11%, and the value decreased with the order of lancing. On the contrary, the codeine and the thebaine content did not change during lancing and the value were 6-7% and 3%, respectively. The papaverine and the noscapine content decreased in the order of lancing. PMID- 14740401 TI - [Studies for analyzing phenoxyethanol and parabens in commercial lotions]. AB - Both phenoxyethanol and parabens are kinds of preservatives in cosmetics and are nominated as the restricted ingredients in cosmetics in Japanese Pharmaceutical Affairs Low. So the analytical methods for phenoxyethanol and parabens were investigated by HPLC. After adding methanol to phenoxyethanol and parabens in lotions, the testing solutions were analyzed by HPLC using the ODS column (CAPCELL PAK C18 column, 4.6 x 250 mm), the mixture either of 50 mmol/l phosphate(pH3.5) and acetonitrile (7:3) for phenoxyethanol or of 50 mmol/l phosphate buffer(pH3.5) and acetonitrile (3:2) for parabens and the detection wavelength of either 270 nm for pehnoxyethanol or 255 nm for parabens. The working curves from 0.5 to 5 micrograms/ml passed through the origin. There were no interference of peaks of phenoxyethanol and parabens from the cosmetics. Eleven lotions out of 42 lotions did not have phenoxyethanol. Phenoxyethanol in thirty-one cosmetics ranged from 0.009 to 1.043% and those average was 0.264%. Parabens, especially methylparaben, were detected in all lotions. The amounts of parabens ranged from 0.016 to 0.267% and their average was 0.120%. There were 7 kinds of lotions whose contents of phenoxyethanol and methylparaben were below 0.1%. PMID- 14740402 TI - [Evaluation of molecular weights of hyaluronate preparations by multi-angle laser light scattering]. AB - Hyaluronate (HA), a glycosaminoglycan polysaccharide, has been used for osteoarthritis, periartritis of the shoulder and rheumatoid arthritis by intraarticular administration, and in ophthalmic surgery such as anterior segment surgery, and eye lotion. In this study, the molecular weight (Mw) of HA preparations were estimated by size-exclusion chromatography (SEC) system consisted of a refractometer (RI) and a multi-angle laser light scattering (MALS). From the results, it has been clarified that a successful characterization of HA samples with Mw up to 2 - 3 x 10(6) g/mol was possible by multidetector system. PMID- 14740403 TI - [Study on a method for delivering scFv recombinant antibody into cultured cells]. AB - We try to develop a method for delivering antibody from blood circulation through blood brain barrier to brain. In order to achieve this goal, antibody has to cross cellular membrane of brain capillary endothelial cells twice. As a first step of our study, we examined the ability for scFv antibody to cross cellular membrane of RBL-2H3 cells once and be delivered into the inside of the cultured cells with the help of TAT peptide. TAT peptide was originally found in Tat protein from the HIV-1 virus and known as one of protein transduction domains. First, oligonucleotide encoding TAT peptide was linked to 5' terminal of gene fragment of scFv antibody by PCR technology. TAT-linked scFv gene fragment was subcloned into pET-23b vector and successfully expressed in E. coli as inclusion body. After solubilization and purification, TAT-linked scFv recombinant protein was added to the culture of RBL-2H3 cells. TAT-linked scFv delivered into RBL-2H3 cells was detected by means of immunocytochemistry using fluorescence microscopy. TAT-linked scFv crossed cellular membrane more efficiently than scFv without TAT peptide. PMID- 14740404 TI - [Twenty-eight day repeated dose oral toxicity test of synergist of a pyrethroid insecticide, 2,3,3,3,2',3',3',3'-Octachlorodipropyl ether (S-421) in rats]. AB - 2,3,3,3,2',3',3',3'-Octachlorodipropyl ether (Abbreviation; S-421) is originally developed as synergist of a pyrethroid insecticide. In recent years, S-421 is used widely at home, for a mosquito-repellent incense, electric mosquito repellent, an insect-killing spray, a vacuum cleaner paper pack, etc. as well. On the other hand, S-421 has been detected in vacuum cleaner dust samples as well as human milk samples in Japan indicating that our living environment is already contaminated by this compound. Long term toxicity studies including a carcinogenesis study have been performed and NOEL of chronic toxicity has been settled. However, it is clear that S-421 is used in close proximity so that acute or subacute exposure at relatively higher dose levels than chronic NOEL values are easily assumed, such as use of a spray in an ill-ventilated room, etc. This study, 28 day repeated oral dose toxicity study of S-421 was performed to monitor the outcome of acute and subacute exposure assuming possible exposure accidents mentioned above. The protocol is as follows; Groups of 10 rats of each sex(5 week old), were treated with intragastric administration of S-421 with a dose of 0 (olive oil, control), 10, 40, 160 or 640 mg/kg body weight. For recovery test, 14 day after the last treatment, the control and 640 mg/kg groups were examined, respectively. All animals of all groups in both sexes survived. In the 640 mg/kg groups of the both sexes, all animals were set to drowsiness from about 5 hours after administration, however, they recovered by the next morning. In the hematology examination, Hb, MCH, MCHC, WBC values were significantly decreased and MCV value was significant increased in the 640 mg/kg group of both sexes. In the serum biochemistry, items increased in the 640 mg/kg groups of both sexes returned to normal level after 14 days recovery period. Absolute and relative liver weight increase seen in the 160 mg/kg and above also returned to control level after recovery. Histopathologically, slight hepatocellular swelling was observed in the 160 mg/kg groups and severe hepatocellular swelling with vacuolization and slight necrosis was seen in the 640 mg/kg group. In conclusion, the no-observed-effect levels (NOEL) of S-421 under these conditions was judged to be 40 mg/kg/day. PMID- 14740405 TI - [Determination of six p-hydroxybenzoic acid esters in laver (nori) by HPLC]. AB - A rapid and simple method was developed for simultaneous determination of methyl, ethyl, isopropyl, npropyl, isobuthyl and n-buthyl p-hydroxybenzoic acid esters (PHBA-Es) in laver by HPLC. Six PHBA-Es were extracted from laver with n-hexane ethyl acetate (1:1) by shaking. The extract was evaporated. The residue was dissolved in methyl alcohol and determined by HPLC. Recoveries of six PHBA-Es spiked in laver were 93.6-101.2% at the level of 2 micrograms/g. PMID- 14740406 TI - [Estimated production by the official inspection of tar colors (including aluminum lakes) in fiscal year 2002]. AB - There were 157 official inspections of tar colors and their lakes in fiscal year 2002, and all of their samples were qualified. Total production amount of tar colors that passed inspection in Japan in fiscal year 2002 reached 113.6 tons. Tar color production amounts were described by month and by manufacturer. The food tar color produced in the largest amount was Food Yellow No. 4, accounting for 42.5% during this period. PMID- 14740407 TI - [Glycyrrhizinic Acid Reference Standard (Control 031) of National Institute of Health Sciences]. AB - The raw material of glycyrrhizinic acid was examined for preparation of the "Glycyrrhizinic Acid Reference Standard". The analytical data obtained were: UV spectrum: lambda max, 251 nm; and specific absorbance (E(1 cm)1%) in ethanol at 251 nm, 146.4; IR spectrum, specific absorptions of raw material were consistent with those of Standard (Control 0221). Also, thin-layer chromatography, no impurity was detected; high-performance liquid chromatography, several impurities were detected. The amount of each impurity was estimated at less than 0.2% and total amount of impurities was less than 0.2%. Based on the above results, the candidate material was authorized as the Glycyrrhizinic Acid Reference Standard (Control 031) of the National Institute of Health Sciences. PMID- 14740408 TI - [Thiamine Hydrochloride Solution Reference Standard (Control 021) of National Institute of Health Sciences]. AB - The raw material of thiamine hydrochloride was examined for preparation of the "Thiamine Hydrochloride Reference Standard". The analytical data obtained were: assay by HPLC, 100.5%; spectrophotometric assay 101.2%. Based on the above results, the candidate material was authorized as the Thiamine Hydrochloride Reference Standard (Control 021) of the National Institute of Health Sciences. PMID- 14740409 TI - [Paeoniflorin Reference Standard (Control 021) of National Institute of Health Sciences]. AB - The raw material of paeoniflorin was examined for the preparation of the "Paeoniflorin Reference Standard". The analytical data obtained were: UV spectrum, lambda max, 231.9 nm; and specific absorbance (E(1 cm)1%) in methanol at 230 nm = 263.4 IR spectrum, specific absorptions of the raw material were consistent with those of the Paeoniflorin Reference Standard Standard (Control 012); High-performance liquid chromatography(HPLC), several impurities were detected. The total amount of impurities was 0.86%. Based on the above results, the candidate material was authorized as the Paeoniflorin Reference Standard (Control 021) of the National Institute of Health Sciences. PMID- 14740410 TI - [Estrone Reference Standard (Control 021) of National Institute of Health Sciences]. AB - The raw material of estrone acetate was examined for the preparation of the "Estrone Reference Standard (Control 021)", The analytical data obtained were: UV spectrum, lambda max of 281 nm and specific absorbance in ethanol at 281 nm = 76.1; Melting point, 261.6 degrees C; IR spectrum, exhibited the specific absorptions at 3345, 2867, 1720, 1499 and 1055 cm-1; thin-layer chromatography, 2 impurities were detected at 100 and 200 micrograms; high-performance liquid chromatography, total amount of impurities estimated to be less than 0.3%. Based on the above results, the raw material was authorized as Estrone Reference Standard (Control 021) of the National Institute of Health Sciences. PMID- 14740411 TI - [Cholecalciferol Reference Standard (Control 031) of National Institute of Health Sciences]. AB - The raw material of cholecalciferol was examined for the preparation of the "Cholecalsiferol Reference Standard (Control 031)", The analytical data obtained were: melting point, 86.3 degrees C; UV spectrum, lambda max of 264.6 nm and specific absorbance in ethanol at 265 nm = 483.6; IR spectrum, same as that of the cholecalciferol Reference Standard (Control 003); optical rotation [alpha]20(D) = 105.1 degrees; thin-layer chromatography, one impurity was detected at 50 micrograms; high-performance liquid chromatography, total amount of impurities estimated to be less than 0.04%. Based on the above results, the raw material was authorized as the Japanese Pharmacopoeia Cholecalciferol Reference Standard (Control 031) of the National Institute of Health Sciences. PMID- 14740412 TI - [Tocopherol Acetate Reference Standard (Control 021) of National Institute of Health Sciences]. AB - The raw material of tocopherol acetate was examined for the preparation of the "Tocopherol Acetate Reference Standard (Control 021)", The analytical data obtained were: UV spectrum, lambda max of 278.5 and 284.8 nm and specific absorbance in ethanol at 284 nm = 42.9; IR spectrum, same as that of the Tocopherol Acetate Reference Standard (Control 001); thin-layer chromatography, no impurities were detected until 50 micrograms; high-performance liquid chromatography, total amount of impurities estimated to be less than 0.6%. Based on the above results, the raw material was authorized as the Japanese Pharmacopoeia Tocopherol Acetate Reference Standard (Control 021) of the National Institute of Health Sciences. PMID- 14740413 TI - [Ergocalciferol Reference Standard (Control 031) of National Institute of Health Sciences]. AB - The raw material of ergocalciferol was examined for the preparation of the "Ergocalsiferol Reference Standard (Control 031)", The analytical data obtained were: melting point, 114.5 degrees C; UV spectrum, lambda max of 264.8 nm and specific absorbance in ethanol at 265 nm = 474.7; IR spectrum, same as that of the Ergocalciferol Reference Standard (Control 003); optical rotation, [alpha]20(D) = +104.6 degrees ; thin-layer chromatography, two impurities were detected at 50 micrograms; high-performance liquid chromatography, total amount of impurities estimated to be less than 0.04%. Based on the above results, the raw material was authorized as the Japanese Pharmacopoeia Ergocalciferol Reference Standard (Control 031) of the National Institute of Health Sciences. PMID- 14740414 TI - [Thrombin Reference Standard (Control 031) of National Institute of Health Sciences]. AB - The "Thrombin Reference Standard (Control 031", of National Institute of Health Sciences was prepared. The precision of filling into ampoule was about 11.5% as C.V. The content of alpha-thrombin was about 89%. The thrombin potency of the standard material was assayed against the Thrombin Reference Standard (Control 961) according to the method of JP-X IV and the potency was 692 +/- 35 units/ampoule. From the results, the potency of the proposed material for Thrombin Reference Standard was defined as 690 units per ampoule. PMID- 14740415 TI - [Lysozyme Reference Standard (Control 031) of National Institute of Health Sciences]. AB - The "Lysozyme Reference Standard (Control 951031)" of the National Institute of Health Sciences was prepared. The lysozyme potency of the standard material was assayed against the Lysozyme Reference Standard (Control 951) by turbidimetric method two turbidimetric methods using the dried y-cells of Micrococcus luteus as a substrate. The potency of the standard material was in satisfactory agreement with that of Lysozyme Reference Standard (Control 951) and was defined as 1 mg [potency] per mg. PMID- 14740416 TI - [Studies on flavianic acids as color reference standard for thin-layer chromatography]. AB - Various flavianic acids were prepared and examined for the preparation for color reference standard for thin-layer chromatography (TLC). Their analytical data were: IR spetra, their specific absorptions were same; no impurities were detected; specific odor-free. It was clear that the prepared flavianic acids were useable as color reference standards for TLC. PMID- 14740417 TI - [Studies on eight color reference standards for thin-layer chromatography]. AB - Eight color NIHS reference standards were studied on IR spectra and thin-layer chromatography (TLC). Their IR spectra and TLCs were determined. They were also authorized as reference standards for TLC. PMID- 14740420 TI - [Potential role of regulatory science in the development of novel biologics]. PMID- 14740421 TI - Do we need 18F-FDG-positron emission tomography as a functional imaging technique for diagnosing large vessel arteritis? PMID- 14740422 TI - Mass spectrometry-based proteomics and analyses of serum: a primer for the clinical investigator. AB - The vocabulary of proteomics and the swiftly-developing, technological nature of the field constitute substantial barriers to clinical investigators. In recent years, mass spectrometry has emerged as the most promising technique in this field. The purpose of this review is to introduce the field of mass spectrometry based proteomics to clinical investigators, to explain many of the relevant terms, to introduce the equipment employed in this field, and to outline approaches to asking clinical questions using a proteomic approach. Examples of clinical applications of proteomic techniques are provided from the fields of cancer and vasculitis research, with an emphasis on a pattern recognition approach. PMID- 14740423 TI - The use of (18F)fluoro-deoxyglucose positron emission tomography in the assessment of large vessel vasculitis. AB - FDG-PET scan has recently been introduced as a diagnostic means to assess large vessel involvement in giant cell arteritis (GCA). Its use in Takayasu arteritis, idiopathic periaortitis and multifocal fibrosclerosis--although more limited, due to the relative rarity of these conditions compared to GCA--is discussed as well. PMID- 14740424 TI - Laboratory investigations useful in giant cell arteritis and Takayasu's arteritis. AB - A raised erythrocyte sedimentation rate (ESR) is considered a hallmark for the diagnosis of giant cell arteritis (GCA). The American College of Rheumatology 1990 criteria for GCA include ESR greater than or equal to 50 mm/h as one of the five criteria. Although the presence of a normal ESR made GCA less likely, the results of a population-based study showed that the occurrence of a low/normal value in GCA at diagnosis is not rare. Pre-treatment ESR may be a prognostic indicator for duration of treatment. C-reactive protein (CRP) and interleukin-6 (IL-6) may be more sensitive indicators of disease activity than ESR in GCA patients. However, it is unclear whether the use in clinical practice of CRP and IL-6 has some apparent advantage over ESR. ESR is the most often used tool to assess disease activity in Takayasu's arteritis (TA). However, some studies have found that ESR and CRP are not able to differentiate patients with clinically active and inactive TA. Furthermore, histopathological studies have shown that over 40% of patients thought to be in clinical remission with normal acute phase reactants have active arteritis. IL-6 could be a promising marker of disease activity in TA; however, further studies are required to confirm its usefulness in clinical practice. Other laboratory investigations could be useful in the diagnosis or follow-up of GCA and TA, but more studies are required. PMID- 14740425 TI - The role of disease-modifying antirheumatic drugs in the treatment of giant cell arteritis. AB - Giant cell arteritis (GCA) is a vasculitis of unknown etiology that affects medium-sized and large arteries, and is the most common form of vasculitis in populations of predominantly Northern European ancestry. If left untreated, GCA can lead to significant morbidity including blindness, stroke, aortic aneurysm and dissection, as well as large-artery stenosis. Glucocorticosteroids are in general very effective in the treatment of GCA. Treatment with high dose glucocorticosteroids is associated with considerable morbidity and for some, but not all patients is required for prolonged periods sometimes exceeding several years. Numerous efforts have been made over the decades to optimize therapeutic outcomes and reduce the side effects of glucocorticosteroids by enlisting adjuvant and alternative therapies. This review focuses primarily on evidence from randomized controlled trials with the objective of efficacy assessment of the respective drugs and dosing regimens in the treatment of GCA. Various glucocorticosteroid dosing regimens including alternate-day dosing, disease modifying therapies including methotrexate and azathioprine, and the prospect of using monoclonal anti-cytokine or anti-cytokine receptor antibodies in the treatment of GCA are addressed. Thus far, no disease-modifying antirheumatic drug therapy or alternative to daily glucocorticosteroid therapy has been found to be unequivocally effective in the treatment of GCA. PMID- 14740426 TI - New developments in the pathogenesis of ANCA-associated vasculitis. AB - In recent years there have been substantial developments in the understanding of the pathogenesis of ANCA-associated vasculitidies. Animal models have now been developed that finally prove a direct pathogenic role for ANCA, a subject fiercely debated since their original identification. We are also closer to understanding how ANCA exert their effects to cause disease. Progress has been made in elucidating how ANCA activate neutrophils, from how they bind antigen and where that antigen is located, to how antigen binding is translated into intracellular activity. The effects of ANCA activation on the effector functions of neutrophils and monocytes are being further dissected and the flow-based assay is allowing interactions with endothelium to be studied in more detail. Knowledge of the role of T cells has been enhanced by examining contributions to disease by differing subsets and their cytokine secretions. Defects in apoptosis playing a role in the initiation of other autoimmune diseases has prompted investigations into whether a similar pathogenesis is relevant in vasculitis, and various genetic polymorphisms have been discovered to be important in determining in whom vasculitis develops. This article reviews how recent research has helped in the understanding of the pathogenesis of small vessel vasculitis. PMID- 14740427 TI - Immune phenomena in localized and generalized Wegener's granulomatosis. AB - Wegener's granulomatosis (WG) is characterized by granulomatous inflammation and systemic vasculitis with a predilection for the lungs and kidneys. In most patients WG begins with a localized organ involvement of the upper respiratory tract that progresses to systemic disease (generalized WG) (1). Because of the life-threatening nature of systemic vasculitis, much effort has concentrated on elucidating the pathogenesis of the vasculitis. However, based upon a renewed interest in (innate) immune defenses against microbes, a better understanding of the chronic granulomatous inflammation may contribute to a more precise insight into the early genesis of WG. Thus, this review focuses on summarizing and discussing data for a potential pattern of disease, i.e. from localized to generalized WG with a special emphasis on granulomatous lesions of the upper respiratory tract and their alterations during the disease course. PMID- 14740428 TI - The role of myeloperoxidase in the pathogenesis of systemic vasculitis. AB - Wegener's granulomatosis, microscopic polyangiitis, Churg-Strauss syndrome and idiopathic pauci-immune necrotizing crescentic glomerulonephritis are strongly associated with the presence of anti-neutrophil cytoplasmic antibodies (ANCA). These ANCA-associated vasculitides can serologically be separated into myeloperoxidase (MPO)-ANCA and proteinase 3 (PR3)-ANCA positive patients. The unique properties of the antigen targeted by the anti-MPO antibodies could help to explain the specific characteristics of MPO-ANCA associated disease. Recently, an animal model has been developed that proves that anti-mouse MPO immunoglobulins alone are capable of causing disease similar to that in humans. Also, the in vitro pathologic effects of binding of MPO-ANCA to MPO are better understood. MPO-ANCA can activate (primed) neutrophils directly causing extensive reactive oxygen species formation and degranulation of neutrophil constituents, including MPO, resulting in a destructive inflammatory response towards the vessel wall. MPO-ANCA can prevent the clearing and inactivation of MPO by ceruloplasmin as well, resulting in increased myeloperoxidase activity. Myeloperoxidase produces not only the strong oxidant bleach (hypochlorous acid) out of hydrogen peroxide and chloride ions but also oxidizes LDL into a macrophage high-uptake form, inactivates protease inhibitors, and consumes nitric oxide. These may contribute to endothelial dysfunction and add to the chronic renal lesions observed in patients with MPO-ANCA. MPO levels are influenced by genetic factors including two, MPO463 and MPO129, single nucleotide polymorphisms. The MPO 463 polymorphism has been associated with an increased risk of development of MPO-ANCA associated disease. PMID- 14740429 TI - Membrane proteinase 3 expression on resting neutrophils as a pathogenic factor in PR3-ANCA-associated vasculitis. AB - Antineutrophil cytoplasm autoantibody (ANCA)-associated small vessel vasculitides are systemic diseases characterized by chronic inflammation of blood vessels. These vasculitides are associated with the presence of ANCA which are, in most cases, directed towards proteinase 3 (PR3) or myeloperoxidase (MPO). In vitro and in vivo data have suggested a pathophysiological role in the ANCA-associated vasculitides, particularly based on the capacity of autoantibodies to bind and activate neutrophils. This review focuses on the role of constitutive expression of proteinase 3 on the membrane of resting neutrophils (mPR3). mPR3 can be expressed on the total population or on a subset of neutrophils and levels of mPR3 differ between individuals. The level of mPR3 on resting neutrophils and the percentage of mPR3 expressing neutrophils is stable in time for a given individual, suggesting a genetic determinant. Patients with ANCA-associated vasculitis have an increased constitutive expression of mPR3 on resting neutrophils compared to healthy controls. High levels of mPR3 on resting neutrophils are a risk factor for the development of relapses in patients with PR3-ANCA-associated vasculitis, probably by making resting neutrophils more susceptible for binding ANCA and induction of activation. As such, constitutive mPR3 expression on neutrophils seems another pathogenic factor in ANCA-associated vasculitis. PMID- 14740430 TI - Update on the pathogenesis of Churg-Strauss syndrome. AB - Churg-Strauss syndrome (CSS) is a rare form of systemic vasculitis occurring in patients with asthma. The cause of CSS is unknown, and yet little data are available regarding its pathogenesis. The presence of a marked tissue- and blood eosinophilia, as well as secretory products of eosinophils in blood and tissues, implicates a pathogenetic role of eosinophil granulocytes. Prolonged survival of eosinophils due to inhibition of CD95-mediated apoptosis by soluble CD95 seems to contribute to eosinophilia in CSS. Although the mechanisms involved in eosinophil activation in CSS have not been elucidated, recent data suggest a possible role of T lymphocytes secreting eosinophil-activating cytokines. This review describes the current insights into the pathogenesis of CSS in the light of its putative nature as a type 2 granulomatous disease. Recent clinical, experimental and epidemiologic data regarding the possible role of inflammatory cells and their secretory products, anti neutrophil cytoplasm antibodies (ANCA), epidemiologic factors and anti-asthma treatments are summarized. PMID- 14740431 TI - HCV-related cryoglobulinemic vasculitis: an update on its etiopathogenesis and therapeutic strategies. AB - Cryoglobulinemic vasculitis (CV) is an immune-complex-mediated systemic vasculitis involving small-medium sized vessels. A causative role of hepatitis C virus (HCV) in over 4/5 patients has been definitely established on the basis of epidemiological, pathological, and laboratory studies. There is great geographical heterogeneity in the prevalence of CV as well as other HCV-related immuno-lymphoproliferative disorders. Thus, unknown environmental and/or genetic co-factors should contribute to the pathogenesis of these conditions. Due to the biological properties, HCV genomic sequences cannot be integrated into the host genome; the virus could trigger the immunological alterations only indirectly by exerting a chronic stimulus to the immune system. Recent laboratory observations gave us new important insights on the complex pathogenetic mechanism(s) of HCV related CV. Firstly, the HCV envelop protein E2, able to bind CD81 molecule expressed on B-lymphocytes, might be involved in the first steps of HCV-driven autoimmune and lymphoproliferative phenomena. The interaction between HCV-E2 and CD81 may increase the frequency of VDJ rearrangement in antigen-reactive B-cell. One possible consequence may be the activation of anti-apoptotic Bcl-2 protoncogene that leads to extended B-cell survival. Interestingly, t(14, 18) translocation along with Bcl-2 activation have been demonstrated in B-lymphocytes of 80% HCV-related CV. The B-lymphocyte expansion is responsible for a wide autoantibody and immune-complex production, including mixed cryoglobulins. CV shows a relatively benign clinical course; however, its cumulative survival is significantly worse if compared to general population. For a correct therapeutic approach to HCV-related CV we must deal with conflicting conditions: HCV infection, autoimmune, and lymphoproliferative alterations. Therapeutic strategy of CV includes etiologic, pathogenetic, and/or symptomatic therapies, which should be tailored for the single patient according to the severity of clinical symptoms. A careful clinical monitoring of patients with HCV-related CV is mandatory in all cases, with particular attention to neoplastic complications. PMID- 14740432 TI - Cutaneous vasculitis: a diagnostic approach. PMID- 14740433 TI - Prevalence of ANCA in mixed cryoglobulinemia and chronic hepatitis C virus infection. AB - OBJECTIVE: To determine the prevalence, target antigens and clinical associations of antineutrophil cytoplasmic antibodies (ANCA) in chronic hepatitis C without extrahepatic manifestations and in chronic hepatitis C virus (HCV)-associated mixed cryoglobulinemia (MC) in two European centers. METHODS: 50 sera from patients with chronic hepatitis C and 116 sera from HCV-associated MC were tested for cytoplasmic or perinuclear pattern (C-ANCA/P-ANCA) by indirect immunofluorescence test (IFT). ANCA target antigens were determined by enzyme linked immunosorbent assay (ELISA). RESULTS: Clinical characteristics of the patients were not different between the two centers. Cryoglobulinemic vasculitis (CV) was biopsy-proven in about 90% of the MC patients. Two patients with HCV associated MC and 1 patient with chronic hepatitis C had a P-ANCA. A C-ANCA was detected in 1 patient with HCV-associated MC. Eight patients with a HCV associated MC and 5 patients with chronic hepatitis C had an ANCA either directed against bactericidal/permeability increasing protein (BPI) or cathepsin G (CG). BPI- or CG-ANCA positivity was not associated with a more severe disease course. The C-ANCA titer followed disease activity in one C-ANCA positive HCV-associated MC patient. The subspecificity of the C-ANCA was not determinable in that patient. CONCLUSION: Two new target antigens of ANCA have been identified in HCV associated MC and chronic hepatitis C in this study. BPI-ANCA and GC-ANCA were present in about 10% of patients with HCV-associated MC or chronic hepatitis C. ELISA proved to be more sensitive in the detection of ANCA than IFT. The present study on chronic HCV infection adds to various reports on the induction of CG- and BPI-ANCA in chronic infections. PMID- 14740434 TI - Pseudomonas-induced lung damage in cystic fibrosis correlates to bactericidal permeability increasing protein (BPI)-autoantibodies. AB - OBJECTIVE: Lung damage is the most common cause of death in cystic fibrosis (CF). It is induced by bacterial colonization and inflammatory activity perpetuates its course. Autoantibodies directed against BPI (bactericidal permeability increasing protein), called BPI-ANCA, have recently been associated with cystic fibrosis. Here we confirm this association and evaluate the relation between ANCA and total IgG level as they relate to bacterial colonization, pulmonary function, and musculoskeletal symptoms. METHODS: BPI-ANCA, MPO-ANCA, and PR3-ANCA were measured with ELISA in 46 adult patients with CF. Total IgG was determined by immunoturbidimetry. Results were correlated to bacterial colonization, lung function and musculoskeletal symptoms. RESULTS: BPI-ANCA was found in 33 patients. In the whole group, both BPI-ANCA and total IgG were inversely correlated to lung function, but in patients chronically colonized with Pseudomonas aeruginosa (P. aeruginosa), BPI-ANCA alone was correlated to lung damage (p = 0.01). Median lung function, measured as forced expiratory volume in 1 second, in P. aeruginosa colonized patients with high levels of BPI-ANCA was 43% of the predicted value. In BPI-ANCA negative, the corresponding figure was 83%. In patients not colonized with P. aeruginosa, this relation was less evident. No correlation between ANCA and musculoskeletal symptoms was seen. CONCLUSION: P. aeruginosa induced lung damage in CF patients is associated with the presence of BPI-ANCA. P. aeruginosa colonized patients without BPI-ANCA have almost normal lung function. We suggest that BPI-ANCA discriminate P. aeruginosa colonized CF patients with severe lung damage from those whose disease is less destructive. Vasculitis like symptoms in CF are not ANCA associated. PMID- 14740435 TI - Study of host and virological factors of patients with chronic HCV infection and associated laboratory or clinical autoimmune manifestations. AB - OBJECTIVE: Chronic hepatitis C virus (HCV) infection is associated with an array of autoimmune laboratory and clinical manifestations. The goals of our study were to identify host and/or virological factors that are implicated in the pathogenesis of these manifestations. METHODS: We performed a detailed prospective study of various demographic, virological, biochemical, immunological (including lymphocyte subsets, Fc gamma-receptor and HLA class-II genotyping), histological and host genetic parameters in 3 well defined subgroups of HCV patients (n = 40): patients with liver disease only (group I, n = 11) or with laboratory (group II, n = 20) and clinical (group III, n = 9) autoimmune manifestations. RESULTS: Group III patients, mainly with features of mixed cryoglobulinemia, were older, with higher levels of rheumatoid factor and circulating cryoglobulins while they tended to have a longer estimated disease duration compared to the other two groups of patients. We did not identify any specific immunological features that could differentiate symptomatic versus asymptomatic patients, except from the elevated soluble interleukin-2 receptor levels. An increased frequency of the R/R131 FcR gamma IIIA and the NA1/NA1 Fc gamma RIIIB genotypes was observed in our total HCV population, regardless of autoimmune manifestations, compared to historical controls. No statistically significant differences in HLA class II allele frequencies was detected between patient subgroups or in comparison to healthy controls. CONCLUSIONS: Chronically infected HCV patients with symptomatic mixed cryoglobulinemia display a number of unique characteristics that differentiate them from asymptomatic patients with chronic hepatitis C. PMID- 14740436 TI - Analysis of the clinical profile, autoimmune phenomena and T cell subsets (CD4 and CD8) in Takayasu's arteritis: a hospital-based study. AB - OBJECTIVE: To evaluate clinical and immunological abnormalities in patients with Takayasu's arteritis (TA) from India, with particular reference to autoimmune perturbations and abnormalities in T cell subsets (CD4 and CD8 cells). METHODS: 16 consecutive patients with TA (11 females and 5 males) underwent clinical and laboratory evaluation inclusive of flow cytometric analysis of T cell subsets (CD4 and CD8). A control population of 94 age- and sex-matched blood donors was used to determine the normal T cell subsets. Student's t-test was used to compare the means. RESULTS: The mean age at onset was 23.4 + 2.3 yrs. Common symptoms observed were headache, limb claudication, abdominal pain and visual disturbance/blackout. Common clinical signs observed included reduced arterial pulsations, bruits, and a BP difference > 10 mm Hg in the upper limbs. Systemic hypertension was documented in 12 patients. The mean absolute lymphocyte count in the patients was 2289/mm3. The mean CD4 count and CD4% were 1003 and 41 respectively; the mean CD8 count and CD8% were 755 and 34, respectively; and the mean CD4/8 ratio was 1.41. The patients had statistically significantly higher CD8 but not CD4 T cell values than controls. IgG and IgM immunoglobulin levels were increased. The mean multi-test CMI score in patients using CMI multi-test device of Pasteur Merieux was 14.6 mm. Two patients had an anergic response, 4 a partial response (1-13 mm), and 6 a full response of > 13 mm. Four patients hyper responded with a score of > 20 mm. ANCA was positive in 2 patients. ANA was positive in 3 patients. IgG anticardiolipin was positive in 12 patients and IgM in 3; overall 12 patients were anticardiolipin positive by ELISA. Anti-beta 2GPI of the IgG variety was found to be positive in 3 patients and IgM in 2 patients; overall 3 patients being positive for the same. Nine of the patients with active disease were started on a combination of moderate dose prednisolone (20-40 mg once daily) along with weekly oral methotrexate (7.5-15.0 mg). Surgical intervention was required in 6 patients. CONCLUSION: This study found an increase in CD8 positive T cell subsets, increased IgG and IgM immunoglobulin levels, and the presence of autoantibodies including ANA, ANCA, anticardiolipin and anti-beta 2GPI antibodies in TA patients. TA may be an autoimmune disorder with T cell aberrations. The relationship with antiphospholipid antibodies and anti-beta 2GPI needs to be explored and confirmed by other larger studies. The strikingly positive responses to tuberculin, as well as the multi-test CMI also indicate exaggerated T cell responses and cell mediated immunity in Takayasu's arteritis. Immunosuppressive therapy was successful in controlling disease activity in the majority, but surgery was needed for irreversible stenotic lesions. PMID- 14740437 TI - ANCA against bactericidal/permeability-increasing protein, azurocidin, calprotectin and defensins in rheumatic and infectious diseases: prevalence and clinical associations. AB - OBJECTIVE: To determine the prevalence and clinical associations of ANCA against the antibiotic proteins and peptides: Bactericidal/permeability-increasing protein (BPI), Azurocidin (AZ), Calprotectin (CP) and beta-Defensin-1 and -2 (DF). METHODS: Patients with ANCA-associated vasculitides (n = 99), other vasculitides and rheumatic connective tissue diseases (n = 303), HIV-infection (n = 66), other infectious diseases (n = 134) Crohn's disease (n = 12) and ulcerative colitis (n = 12) were tested for BPI-, AZ-, CP-, DF-, PR3-, and MPO ANCA in indirect immunofluorescence technique (IFT) and ELISA. RESULTS: In ANCA associated vasculitides BPI-ANCA were detected in 6% of patients. In HIV infection, BPI was the main target antigen of ANCA-IFT positive sera (74%). BPI ANCA was associated with higher inflammatory activity. In Crohn's disease and ulcerative colitis BPI-ANCA was prominent (34% of patients). AZ-ANCA were found in 5% of patients. No ANCA were detected against defensin and calprotectin. CONCLUSION: BPI-ANCA is the main autoantibody in HIV and is associated with higher inflammatory activity. In inflammatory bowel diseases BPI-ANCA is predominant, AZ-ANCA are also present to a lesser extend. Both were not useful characterize clinical subgroups. No ANCA were detected against calprotectin or defensins. PMID- 14740438 TI - Visceral leishmaniasis in a patient treated for polyarteritis nodosa. AB - We report a case of visceral leishmaniasis in a patient receiving steroids and cyclophosphamide for polyarteritis nodosa. The clinical presentation of leishmaniasis, with fever, pancitopenia and hypergammaglobulinemia may be confused with a reactivation of vasculitis or with other non-infectious conditions such as haematologic diseases. In endemic areas, the case of latent leishmaniasis must be considered and serology for Leishmania should be performed before starting immunosuppressive treatment. PMID- 14740439 TI - Pulmonary artery involvement in Wegener's granulomatosis. AB - Classification and nomenclature schemes are guidelines and are not intended to recognize and distinguish the entire spectrum of any single disease. It may in fact be misleading to suggest that a classic presentation of a given disease plus atypical features should be considered an "overlap" of two separate, often rare, conditions. We report a case of typical Wegener's granulomatosis (WG), with the coexistence of pulmonary artery stenosis, a lesion more commonly observed in TA. This is not the first or only example of large vessel vasculitis occurring in patients with WG. This observation cautions clinicians to avoid rigid application of classification and nomenclature systems and raises questions about determinants of vasculitis subsets and organ targeting. PMID- 14740440 TI - Clinical management issues in vasculitis. Angiographically defined angiitis of the central nervous system: diagnostic and therapeutic dilemmas. AB - A case of acute neurologic deficit accompanied by a cerebral angiogram consistent with CNS vasculitis is presented. The differential diagnosis and diagnostic decision process generated in this type of evaluation is illustrated. PMID- 14740441 TI - A headache and a mass lesion: vasculitis or CNS sarcoid? PMID- 14740442 TI - Large vessel vasculitis. PMID- 14740443 TI - Successful response to infliximab in a patient with undifferentiated spondyloarthropathy coexisting with polyarteritis nodosa-like cutaneous vasculitis. PMID- 14740444 TI - New roles for estrogens in rheumatoid arthritis. AB - Sex hormones appear to play an important role as modulators of autoimmune disease onset/perpetuation. Steroid hormones are implicated in the immune response, with estrogens as enhancers at least of humoral immunity, and androgens and progesterone (and glucocorticoids) as natural immune suppressors. Serum levels of estrogens have been found to be normal in rheumatoid arthritis (RA) patients. Synovial fluid levels (SF) of proinflammatory estrogens relative to androgens are significantly elevated in both male and female RA patients as compared to controls, which is most probably due to an increase in local aromatase activity. Thus, available steroid pre-hormones are rapidly converted to proinflammatory estrogens in the synovial tissue in the presence of inflammatory cytokines (i.e. TNF alpha, IL-1, IL-6). The increased estrogen concentrations observed in RA SF of both sexes are characterized mainly by the hydroxylated forms, in particular 16 alpha-hydroxyestrone, showing a mitogenic stimulating role. Indeed, recent studies by us indicate that 17-beta estradiol (E2) clearly enhanced the expression of markers of cell growth and proliferation, whereas testosterone (T) induced an increase in markers indicating DNA damage and apoptosis. In particular, our data further shows that the enhancing role of estrogens on the immune/inflammatory response is exerted by activating the NFkB complex. In conclusion, locally increased estrogens may exert activating effects on synovial cell proliferation, including macrophages and fibroblasts, suggesting new roles for estrogens in RA. PMID- 14740445 TI - Guidelines for clinical studies assessing the efficacy of drugs for the management of acute low back pain. AB - In this paper we propose guidelines for clinical trials aimed at assessing the efficacy of drugs for acute non-specific low back pain (LBP) with or without radicular pain, preliminary to their approval and registration. To this end, consensus statements were obtained from a group of experts in the fields of rheumatology, clinical medicine, public health and epidemiology. EBM resources were systematically used as references. Four diagnostic categories were defined: type 1--LBP with no radiation; type 2--LBP radiating no further than the knee; type 3--LBP radiating beyond the knee, but with no neurologic signs; and type 4- LBP radiating to a specific and entire leg dermatome, with or without neurologic signs. Studies should be designed on the basis of the claimed indications for the drug, but must be double-blinded whatever the indication. The duration of the study may be shorter for LBP type 1 or 2 (one week) than for LBP types 3 and 4 (up to one month), depending on the aim of the study and the indications for the drug. The comparator may be inactive (placebo) or active (for a superiority trial, e.g., versus paracetamol). Specific inclusion and exclusion criteria have been defined here for each category. An appropriate wash-out period for any drugs that could affect the pain status should be planned. Paracetamol may be allowed as rescue medication. The primary endpoint should be based on a validated pain assessment tool that may be either generic (type 1 or 2) or oriented (back and knee for types 3 and 4). Secondary endpoints could include the assessment of functional performance; the duration of any period of bed-rest; work limitation; a global assessment comprising pain at rest, standing and walking; the time elapsed before epidural injection, the prescription of other therapeutic agents, or surgery; and the use of rescue medication. Adverse events (AE) should be monitored systematically using a methodology that reflects the mode of action of the tested drug. With the application of these guidelines, LBP could serve as an appropriate disease for testing analgesic drugs. Rigorous evaluation may also help to improve the management of acute LBP. PMID- 14740446 TI - Leflunomide improves the clinical response in patients with active rheumatoid arthritis treated with methotrexate. PMID- 14740447 TI - Autonomic nervous system dysfunction involving the gastrointestinal and the urinary tracts in primary Sjogren's syndrome. AB - OBJECTIVE: Antibodies reacting with the m3 subtype muscarinic acetylcholine receptor appear to be an important pathogenic factor in primary Sjogren's syndrome (pSS). As this receptor subtype is functionally important in the gastrointestinal and urinary tracts, and very little is known about the autonomic nervous system function in these organs in pSS patients, the occurrence and clinical significance of an autonomic nervous system dysfunction involving the gastrointestinal and urinary tracts were investigated. METHODS: Data on clinical symptoms attributable to an autonomic dysfunction were collected from 51 pSS patients. Gastric emptying scintigraphy and urodynamic studies were performed on 30 and 16 patients, respectively, and the results were correlated with patient characteristics and with the presence of autonomic nervous system symptoms. RESULTS: Gastric emptying was abnormally slow in 21 of the 30 examined patients (70%). Urodynamic findings, compatible with a decreased detrusor muscle tone or contractility were found in 9 of the 16 patients tested (56%). Various symptoms of an autonomic nervous system dysfunction were reported by 2-16% of the patients. CONCLUSION: Signs of an autonomic nervous system dysfunction involving the gastrointestinal and the urinary systems can be observed in the majority of pSS patients. This high occurrence is rarely associated with clinically significant symptoms. The authors presume a role of autoantibodies reacting with the m3 muscarinic acetylcholine receptor in the elicitation of the autonomic dysfunction. PMID- 14740449 TI - Active MMPs captured by alpha 2 macroglobulin as a marker of disease activity in rheumatoid arthritis. AB - OBJECTIVE: The aim of the present study was to analyze alpha 2 Macroglobulin/MMP (alpha 2M/MMP) complex formation and to investigate whether MMP activity in alpha 2M/MMP complexes in serum can be used as a disease marker in rheumatoid arthritis (RA). METHODS: High and low molecular weight (H/LMW) substrates and inhibitors and size exclusion were used to analyze alpha 2M/MMP complex formation. LMW fluorogenic substrates were used to quantify the level of MMPs in alpha 2M/MMP complexes in the serum of RA patients and healthy controls. RESULTS: Active MMPs were fully inhibited by LMW inhibitor BB94 in the presence of alpha 2M, whereas no inhibition was achieved by HMW inhibitor TIMP-1. Size exclusion analysis showed alpha 2M/MMP complex formation in buffer and in normal plasma spiked with activated MMPs, which indicated alpha 2M/MMP complex formation in the systemic circulation. MMP activity in alpha 2M/MMP complexes in the serum of RA patients was significantly higher than in the serum of healthy controls (P < 0.001). MMP activity levels in the serum of RA patients were correlated with ESR (r = 0.72, P < 0.001). CONCLUSION: In the systemic circulation of RA patients, active MMPs form complexes with alpha 2M and can be detected using LMW fluorogenic substrates. MMP activity measurements in serum allow discrimination between RA patients and healthy controls and provide a new tool for the assessment of the disease process in RA. PMID- 14740448 TI - Osteoarthritic articular chondrocytes stimulate autologous T cell responses in vitro. AB - OBJECTIVE: To clarify the presence of specific T cell immune response to autologous chondrocytes in patients with osteoarthritis (OA). METHODS: Peripheral blood mononuclear cells obtained from OA or post-traumatic patients were co cultured with irradiated autologous chondrocytes, and their proliferative response was assessed using 3H-thymidine incorporation. Expression of HLA-class II molecules was also assessed on chondrocytes by immunohistochemistry or flow cytometry. RESULTS: T cell responses to autologous chondrocytes in OA yielded a significantly greater mean stimulation index (6.35 +/- 1.63) compared to controls (1.21 +/- 0.09, p < 0.01). This response was partially blocked by antibodies against HLA class I, class II, CD4 or CD8. Increased expression of HLA-DP, -DQ, and -DR was observed. CONCLUSION: This study showed the autologous T cell stimulating property of OA chondrocytes in vitro. The elucidation of the autoimmune responses may contribute to the understanding of immune-mediated mechanisms in OA. PMID- 14740450 TI - LY309887, antifolate via the folate receptor suppresses murine type II collagen induced arthritis. AB - OBJECTIVE: To examine the effect of LY309887, an inhibitor of glycinamide ribonucleotide formyltransferase in de novo purine biosynthesis on murine type collagen-induced arthritis (CIA). METHODS: CIA was induced by immunization with bovine type II collagen in adjuvant. The expression of folate receptors was examined in dissected synovial tissues and bone marrow cells from arthritic and non-arthritic mice by the semi-quantitative reverse transcription-polymerase chain reaction. LY309887 was administered to CIA mice after the onset of arthritis. Mice were monitored for arthritis index for 21 days. Levels of IgG1 and IgG2a antibodies against bovine type II collagen were measured in sera from CIA mice with or without LY309887 treatment by the enzyme-linked immunosorbent assay. Histologic analyse were also performed in synovial tissues from arthritic joints with or without LY309887 treatment. RESULTS: Levels of mRNA of folate receptor beta (FR-beta) were elevated in arthritic joints from CIA mice, compared with those in nonarthritic joints. The expression of mRNA of FR-beta was dominant in bone marrow cells of CIA mice. The administration of LY309887 suppressed the disease progression of CIA mice as defined by the lower arthritis index, and decreased production of serum IgG1 and IgG2a anti-type II collagen antibody, and the damage to cartilage or bone. CONCLUSION: The administration of LY309887 was effective on CIA mice. It was suggested that LY309887 might be useful in the treatment of rheumatoid arthritis. PMID- 14740451 TI - B cell activation in rheumatoid arthritis patients under infliximab treatment. AB - OBJECTIVE: To determine whether anti-TNF alpha (infliximab) treatment affects B cell activation in patients with rheumatoid arthritis (RA) METHODS: B cell activation was analyzed in fifteen anti-TNF-treated RA patients. CD23 expression was used as a B cell activation marker and was studied before and after three months of infliximab treatment. PBMC were stimulated with anti-CD3 mAb during 18 h and were separated by rosseting into E+ and E-cells. B cells were assessed in E population by double staining with CD19 and CD23. ELISA assays were used to assess both soluble TNF alpha and circulant immune complexes (CIC) containing TNF alpha. We also used B cells from tonsils to establish the relationship between B cell activation and TNF alpha CIC. RESULTS: The proportion of B cells expressing CD23 was higher before infliximab exposure than after treatment (48.3 +/- 16.7 versus 29.5 +/- 12.5, p = 0.007). T-B cell interactions were assessed by means of blocking antibodies to CD154, CD40, CD69, and CD18; these interactions were not specially affected by infliximab treatment. We could demonstrate CIC containing TNF alpha after infliximab treatment, these CIC, similarly to others IgG containing immune complexes, were capable to downregulate CD23 on B cells. CONCLUSIONS: Infliximab treatment in RA downregulates CD23 expression on T-cell activated B cells. This downregulation is connected with the presence of CIC containing TNF alpha. Presumably, the Fc gamma RIIb1 endows IgG-containing immune complexes, as TNF alpha-anti-TNF alpha, with the capacity to regulate B cells and inflammatory cells. PMID- 14740452 TI - Possible role of DNA hypomethylation in the induction of SLE: relationship to the transcription of human endogenous retroviruses. AB - OBJECTIVE: We investigated the contribution of DNA methyltransferase activity to the transcription of human endogenous retroviruses (HERV), which have been reported to be a plausible causative agent for systemic lupus erythematosus (SLE). METHODS: The reverse transcriptase-polymerase chain reaction (RT-PCR) and real-time quantitative-PCR (RQ-PCR) were used. RESULTS: Our results indicated that treatment with 5-aza-deoxycytidine (5-aza C), a demethylating agent, increased the transcription of messenger RNA (mRNA) for HERV clone 4-1 and decreased mRNA for DNA methyltransferase-1 (DNMT-1; methylation-regulating enzyme) in peripheral blood mononuclear cells (PBMC) from normal individuals. Also, transcription of DNMT-1 mRNA in PBMC from patients with SLE was lower than in cells from normal controls. CONCLUSION: DNA hypomethylation seems to play a significant role in the transcription of HERV clone 4-1 and may be related to the pathogenesis of SLE. PMID- 14740453 TI - Morphological and cytoskeletal aspects of cultivated normal and osteoarthritic human articular chondrocytes after cyclical pressure: a pilot study. AB - OBJECTIVE: This study investigated the effect of hydrostatic cyclical pressure on the cell ultrastructure and cytoskeleton of normal and osteoarthritis (OA) human cultivated chondrocytes in vitro. METHODS: The different effects of pressurization with sinusoidal waves at a minimum pressure of 1 MPa, a maximum pressure of 5 MPa and a frequency of 0.25 Hz for 3 hrs on normal and OA chondrocytes were assessed by transmission electron microscopy (TEM), scanning electron microscopy (SEM) and immunoflurescence microscopy (IF). RESULTS: Structural differences exist between normal and OA chondrocytes at the nuclear, cytoplasmic and cytoskeletal level. Pressurization did not alter the normal chondrocytes, but had a beneficial effect on OA chondrocytes, by increasing the number of cell organelles responsible for synthesis activities. IF examination has shown that the distribution of actin protein in normal chondrocytes is polarized on the apical sides of the cellular cytoplasm. However, in OA chondrocytes the signal of the actin protein is not as well defined. Similarly, the localization of the tubulin protein in normal and OA cells also appears to be different. Hydrostatic pressure did not cause any modification in the cytoskeletal organization of the OA chondrocytes. CONCLUSION: This study confirms the different morphology, structure and cytoskeletal aspect of normal and OA chondrocytes and the important role played by pressure on cell morphology. The recovery of OA chondrocytes observed by an increase of cytoplasmic organelles does not seem to involve the cytoskeleton. PMID- 14740454 TI - Agreement among Ayurvedic practitioners in the identification and treatment of three cases of inflammatory arthritis. AB - OBJECTIVE: To conduct a preliminary investigation into the consistency of approach between three Ayurvedic medicine experts on treatments for inflammatory polyarthritis. METHODS: A convenience sample of three experienced Ayurvedic practitioners was recruited. These practitioners independently assessed three subjects with inflammatory polyarthritis for health status, treatment history, and lifestyle, conducted a physical examination, and then independently determined the treatment plan. The treatment plan was recorded on standardized collection forms. The subject examination order was randomized for each practitioner. Following completion of the assessments, a facilitated discussion among the practitioners permitted each to discuss all aspects of the recommended therapies. Proceedings were audio-taped and the content analyzed. RESULTS: All three practitioners agreed upon a unified concept of Ayurvedic disease origin, disease diagnosis, and treatment approach for each patient. Seven specific treatment groupings (i.e. modalities) emerged: diet, exercise, relaxation, analgesic, anti-inflammatory, immune-enhancing, and detoxification/cleansing. Based on the single visit, the practitioners agreed upon 17 of 21 treatment groups for the three patients. CONCLUSION: Despite Ayurvedic medicine's individualized approach, considerable agreement existed among the practitioners studied. The identified Ayurvedic treatment approaches require investigation in a controlled clinical setting. PMID- 14740455 TI - Cooperative induction of 15-lipoxygenase in rheumatoid synovial cells by IL-4 and proinflammatory cytokines. AB - OBJECTIVE: To clarify the role of interleukin-4 (IL-4) in the expression of 15 lipoxygenase (15-LOX), whose metabolities are known to suppress the inflammatory reaction, in freshly prepared rheumatoid synovial cells. METHODS: Adherent synovial cells were prepared by enzymatic digestion of synovia obtained from patients with rheumatoid arthritis (RA). Protein expression of 15-LOX was determined by Western blot analysis. The messenger RNAs of 15-LOX were determined by reverse transcription and the polymerase chain reaction (RT-PCR). RESULTS: Freshly prepared rheumatoid synovial cells did not express 15-LOX at either the mRNA or protein levels. IL-4 induced the protein expression of 15-LOX after 24 hours of culture. Although interleukin-1 alpha (IL-1 alpha) and tumor necrosis factor alpha (TNF alpha), major inflammatory cytokines in rheumatoid synovia, did not induce the expression of 15-LOX, IL-4 and these inflammatory cytokines synergistically enhanced the protein expression of 15-LOX. The synergistic effect was also observed at the level of mRNA. CONCLUSIONS: We demonstrate that IL-4 cooperated with the inflammatory cytokines IL-1 alpha and TNF alpha to enhance the expression of 15-LOX in rheumatoid synovial cells. Since 15-LOX metabolites have potent anti-inflammatory actions, our data suggest that IL-4 might downregulate rheumatoid inflammation via the induction of 15-LOX and its metabolites. PMID- 14740456 TI - Successful treatment of pure red cell aplasia in systemic lupus erythematosus with cyclosporin A. AB - We report a patient with longstanding systemic lupus erythematosus (SLE) who developed pure red cell aplasia (PRCA). This condition is rare in connective tissue diseases and is reported in 32 previous cases of SLE in literature. Our patient recovered, apparently in response to treatment with high dosage of corticosteroids, but relapse occurred when the prednisone dosage was tapered down to 10 mg/day. The patient was successfully treated with cyclosporin A with no recurrence of the disease in the last 2 years. PMID- 14740458 TI - Methotrexate used as a steroid-sparing agent in non-renal chronic Henoch Schonlein purpura. AB - Henoch-Schonlein purpura (HSP) is the most common form of systemic vasculitis in childhood and a small percentage of children develop a chronic form of the disease that often requires prolonged corticosteroid therapy. A detailed case description of a child with non-renal chronic HSP treated with methotrexate to allow for tapering of corticosteroids is presented. This represents the first report of methotrexate use in the treatment of chronic HSP. PMID- 14740457 TI - ANCA against the bactericidal/permeability increasing protein (BPI-ANCA) can compromise the antibiotic function of BPI in a Wegener's granulomatosis patient. AB - A 54-year old Wegener's granulomatosis patient with PR3-ANCA at diagnosis 2 years ago was admitted with a pulmonary relapse and new subglottic stenosis preceded by pulmonary infections. The patient presented with bactericidal/permeability increasing protein (BPI)-ANCA in ELISA whereas at the same time PR3-ANCA had disappeared. Bronchoalveolar lavage revealed pulmonary infection with Gram negative bacteria. After antibiotic treatment, immunosuppression was started with cyclophosphamide and infliximab due to refractory disease. Remission was induced and BPI-ANCA disappeared. A bacterial growth inhibition assay with BPI and the patient's IgG purified during the actual pulmonary relapse showed inhibition of the antimicrobial activity of BPI in vitro, in contrast to IgG from sera taken 2 years before and after remission was induced. The patient's BPI-ANCA recognised the bioactive N-terminal portion of BPI. Thus a possible mechanism is demonstrated for how BPI-ANCA may contribute to a pro-inflammatory setting during the development of a pulmonary relapse in the absence of PR3-ANCA by impeding bacterial clearance. PMID- 14740459 TI - Detection of anti-ribosome antibodies: a long story of lights and shadows. AB - Circulating autoantibodies against ribosomal proteins characterise a subset of patients with systemic lupus erythematosus. Following the identification of three phosphorylated proteins as the main ribosomal autoantigens recognised by these autoantibodies, several studies have been carried out in the last decade to set up a reliable and sensitive method of detecting anti-ribosome autoantibodies and disclosing their possible clinical relevance in the diagnosis and monitoring of symptoms and signs of the disease. Although a number of clinical associations have been proposed, contrasting results have emerged from these investigations. This review analyses the methodological problems linked with the various techniques used to detect anti-ribosome antibodies and provides a critical update of the clinical associations described in lupus patients to date. PMID- 14740460 TI - Growth reconstitution in juvenile idiopathic arthritis treated with etanercept. AB - OBJECTIVE: Growth failure is a leading problem in uncontrolled juvenile idiopathic arthritis. It also affects 10% of patients who are not treated with corticosteroids. The influence of proinflammatory cytokines like interleukin-1 beta, interleukin-6 and tumour necrosis factor on the neuroendocrine axis as well as on the production of insulin-like growth factors (IGFs) has been postulated. The objective of the current study was to evaluate effects of highly active antirheumatic treatment with tumour necrosis factor antagonist on growth retardation. Seven out of 18 patients with refractory juvenile idiopathic arthritis treated with etanercept demonstrated growth retardation leading to short stature. METHODS: Antropometric measurements and disease activity parameters--including the number of swollen and tender joints, morning stiffness, ESR and CRP levels--were monitored monthly during the first year of treatment and every 3 months thereafter. Serum levels of IGF-1 and IFG-BP were measured as well. RESULTS: Upon treatment with etanercept, growth velocity increased from 3.7 +/- 1.2 cm before the beginning of the therapy to 7.6 +/- 1.2 cm in the first year of treatment (p < 0.001). The average length-standard-deviation-score (SDS) increased from -2.4 +/- 1.0 to -1.9 +/- 0.9 after one year and to -1.1 +/- 0.9 after two years (p = 0.05) indicating catch-up growth. Prior to the therapy, serum levels of insulin-like growth factor-1 and of insulin-like growth factor binding protein-3 were within the normal range but increased significantly upon treatment (p < 0.001). An inverse correlation of the IGF-1 serum level to CRP was found. CONCLUSIONS: An intensified anti-inflammatory treatment using etanercept has a beneficial effect on growth in children with a so far uncontrolled inflammatory disease. This effect might be related to the cessation of the inhibitory effect of proinflammatory cytokines on the synthesis of IGF-1 and IGF BP-3 in the liver. Growth failure should be included in the evaluation of antirheumatic treatment. PMID- 14740461 TI - Gender and age differences in pain, coping and health status among children with chronic arthritis. AB - OBJECTIVE: To investigate pain, coping and health status in children with chronic arthritis. Additional aims were to explore gender variations in pain, coping and health status, and the same factors in children under 16 years versus children 16 years or older. METHODS: For 125 children (43 boys) with a median age of 14.1 years (range 10.3-17.8) disability and discomfort were assessed (Childhood Health Assessment Questionnaire, CHAQ). Pain, coping, fatigue, sleeping and satisfaction with life were evaluated (Visual Analogue Scales). The Pain Intensity Scale was used to assess the variation in pain intensity during one week. RESULTS: The CHAQ disability index (DI) was 0.63. In the total sample, girls had larger numbers of affected joints in upper extremities (p = 0.028) and more days with morning pain (p = 0.025) than boys. Boys reported greater perceived ability to reduce pain than girls (p = 0.020). Compliance with treatment was more frequent in girls than in boys (p = 0.029). In children 16 years of age and older, boys reported greater perceived ability to control pain (p = 0.032) and to reduce pain than girls (p = 0.001) and fewer days with worst pain in the mornings (p = 0.013), as well as less pain variations over the period of a day (p = 0.014) and over a period of a week (p = 0.035). CONCLUSIONS: As a group, the children were clearly affected by their arthritis. Pain was related to the severity of disease. Some significant gender and age differences were highlighted, which should be considered in clinical pain management. PMID- 14740462 TI - Distensibility and pulse wave velocity of the thoracic aorta in patients with juvenile idiopathic arthritis: an MRI study. AB - OBJECTIVE: An increased incidence of cardiovascular disease has been found in rheumatic disorders. Changes in the variables of aortic elasticity in patients with juvenile idiopathic arthritis (JIA) were evaluated and their relationship to inflammation, anti-rheumatic drugs and traditional cardiovascular risk factors were investigated in this study. METHODS: Phase contrast MR was performed in 31 patients with JIA and 28 age and sex matched controls to evaluate the aortic distensibility and pulse wave velocity (PWV). Disease activity variables, plasma lipid profile, homocysteine, thyroid hormones, glucose and insulin were assessed in the patients. RESULTS: Eighteen patients had oligoarticular, 6 polyarticular and 7 systemic disease. Distensibility was lower (mean: 10.25; SD: 4.18) and PWV was higher (mean: 3.68; SD: 1.59) in the patients compared to the controls (mean: 13.4; SD: 4.99), (mean: 1.38; SD: 0.54) respectively (p < 0.01). A positive correlation between PWV and age was observed in the patients (rs = 0.47, p < 0.01) and controls (rs = 0.72, p < 0.01), and a negative correlation between distensibility and age in the patients (rs = -0.59, p < 0.01) and controls (rs = 0.63, p < 0.01). No statistically significant correlations were found between distensibility and PWV and metabolic and disease activity parameters. When distensibility and PWV were adjusted for age no significant differences were found between the three subtypes of JIA. CONCLUSION: JIA is associated with increased aortic stiffness that might suggest subclinical atherosclerosis. Early detection and follow-up by non-invasive methods may be useful in the prevention of future cardiovascular disease. PMID- 14740464 TI - Massive hematuria in rheumatoid arthritis: an unusual manifestation of reactive amyloidosis. PMID- 14740463 TI - Prognostic factors and fetal outcomes of lupus pregnancy in Taiwan. PMID- 14740465 TI - Familial adenomatous polyposis coli associated arthritis and vasculitis. PMID- 14740466 TI - A rare case of dermatomyositis associated with immune-complex type glomerulonephritis, idiopathic thrombopenic purpura, pulmonary fibrosis and lung cancer. PMID- 14740467 TI - Project teams put research into practice and cut infection rates. PMID- 14740468 TI - Patient tracking systems reduce phone calls, spot inefficiencies. PMID- 14740469 TI - FDA urges reports on stapler incidents. PMID- 14740470 TI - Who should mark the surgical site? PMID- 14740471 TI - Fine-tuning surgical site verification. PMID- 14740472 TI - Marking the site for spinal surgery. PMID- 14740473 TI - Changing practice for preop hair removal. PMID- 14740474 TI - COX-2 inhibitor improves pain outcomes. PMID- 14740475 TI - Innovative call plan boosts staff morale. PMID- 14740476 TI - ASCs see losses and wins in Medicare bill. PMID- 14740478 TI - Improving pain control for lap choles. PMID- 14740477 TI - Encouraging better postop pain relief. PMID- 14740479 TI - C-terminal sequencing of peptide hormones using carboxypeptidase Y and SELDI-TOF mass spectrometry. PMID- 14740480 TI - Rapid purification of the outer mitochondrial membrane protein Por1p from Saccharomyces cerevisiae. PMID- 14740481 TI - Large-scale yeast transformation in low-percentage agarose medium. PMID- 14740482 TI - Effectiveness and limitations of uracil-DNA glycosylases in sensitive real-time PCR assays. PMID- 14740483 TI - In ovo intraperitoneal administration of bromodeoxyuridine to avian fetuses. PMID- 14740484 TI - Comparison of RiboGreen and 18S rRNA quantitation for normalizing real-time RT PCR expression analysis. PMID- 14740485 TI - pTOC-KR: a positive selection cloning vector based on the ParE toxin. PMID- 14740486 TI - Cost-benefit analysis of a method using diatomaceous earth to purify Tamm Horsfall protein. PMID- 14740487 TI - Design and performance of an electrical stimulator for long-term contraction of cultured muscle cells. AB - Excitability in muscle cells manifests itself as contractility and may be evoked by electrical stimulation. Here we describe an electrical stimulator device applicable to cells seeded on standard multiwell plates and demonstrate how it effectively stimulates synchronous contraction of skeletal muscle C2C12 cells without damaging them. The electrical stimulator of cultured cells (ESCC) consists of two connection cards and a network of platinum electrodes positioned in such way that each well in a row is uniformly stimulated. The ESCC may produce a range of outputs based on the stimulation parameters it receives from a commercial pulse generator and can be placed in a standard cell incubator, allowing for long-term stimulation as required for biochemical and molecular biological assays. We show that a 90-min stimulation of C2C12 myotubes at 50 V, 30 ms of pulse duration, and 3 Hz of frequency enhances glucose metabolism and glycogen mobilization while oppositely modulating the activity ratio of glycogen metabolizing enzymes. Thus, we demonstrate that long-term electrical stimulation of C2C12 myotubes with the ESCC results in contractility and metabolic changes, as seen in exercising muscle. PMID- 14740488 TI - Improved silencing vector co-expressing GFP and small hairpin RNA. AB - Small interfering RNA (siRNA) is a powerful tool for the specific silencing of gene expression. We developed an improved vector, pG-SUPER, that co-expresses green fluorescent protein (GFP) and small hairpin RNA simultaneously to facilitate analysis of silencing at the level of individual cells. As a test system, we analyzed lamin A/C knockdown in HeLa cells. The GFP signal was a reliable reporter (93%-98%) of strong knockdown (approximately 90%) over a wide range of GFP intensities. The GFP reporter made possible the application of fluorescent-activated cell sorting (FACS) to purify the knockdown cell population. Such populations facilitated Western blotting analysis to determine depletion of the target protein. pG-SUPER was also applied to evaluate gene replacement by exogenous genes rendered refractory to siRNA by introducing silent mutations. Recovery of lamin A was linearly correlated to the expression level of the rescue gene. pG-SUPER will expand plasmid-based siRNA applications through the easy and reliable detection of knockdown and rescued cells. PMID- 14740489 TI - Baculovirus expression system for magnetic sorting of infected cells and enhanced titer determination. AB - Recombinant baculoviruses derived from the Autographa californica nuclear polyhedrosis virus (AcNPV) are widely used to express heterologous genes in insect cells, but the use of the baculovirus expression vector system (BEVS) is hampered by slow and tedious procedures for the selection and separation of baculovirus-infected insect cells and for titer determination. Here we developed a new technology based on the bicistronic vector with a fusion protein of the human integral plasma membrane glycoprotein CD4 and green fluorescent protein (GFP) for concomitant expression of target proteins in insect Sf21 cells. Magnetic cell sorting (MACS) technology with anti-CD4 antibody-labeled superparamagnetic beads was used to separate the baculovirus-infected from the noninfected insect cells and therefore to increase the virus titer and to reduce process time. With the herein described use of the MACS-improved baculovirus expression plasmid MACS in baculovirus expression (pMACSiBac-1), we have been able to select the baculovirus-infected insect cells at an early time point of the infection cycle and therefore enrich the virus titer dramatically. Furthermore, simple end point dilution and GFP fluorescence detection can be used for early and facile detection of recombinant viruses and simplified titer determinations. We show that the bicistronic pMACSiBac-1 with an additional multiple cloning site under the control of the very late promoter polyhedrin (PPH) allows for the expression of target proteins in high amounts, less workloads, and shorter timelines. PMID- 14740490 TI - Selection of appropriate control genes to assess expression of tumor antigens using real-time RT-PCR. AB - Real-time reverse transcription PCR (RT-PCR) is a sensitive and accurate method to monitor gene expression and is often used to profile the expression of putative tumor antigens in the context of immunotherapy. However, this technique consists of several steps, including cell processing, RNA extraction, RNA storage, assessment of RNA concentration, and cDNA synthesis prior to PCR. To compensate for potential variability introduced in this procedure, the expression of housekeeping genes is commonly assessed in parallel with the expression of the gene of interest. In this study, the expression of a variety of housekeeping genes in a panel of 26 different human tumor and embryonal cell lines was assessed using real-time RT-PCR. For some control genes, the variability in expression was significant between different cell lines, despite the equalization of quantities of input RNA. The greatest variability was found for GAPDH. The lowest variability was found for beta-glucuronidase (GUS) and 18S rRNA. While real-time RT-PCR is a powerful tool for gene expression analysis, these results suggest that the choice of control genes to normalize the expression of the gene of interest is critical to the interpretation of experimental results and should be tailored to the nature of the study. PMID- 14740491 TI - Tissue microarrays. AB - The identification of disease-related genes is a major focus of modern biomedical research. Recent techniques, including array-based platforms for molecular profiling of disease tissues such as DNA arrays for expression profiling or matrix comparative genomic hybridization, allow for the comprehensive screening of the whole genome in a single experiment. Consequently, thousands of candidate genes have already been identified that may be linked to disease development and progression, and the process of lead discovery continues unimpeded. The evaluation of the clinical value of such leads is challenging because thousands of well-characterized tissue specimens must be analyzed. Tissue microarray (TMA) technology enables high-throughput tissue analyses to keep pace with the rapid process of lead discovery. With this technique, up to 1000 minute tissue samples are brought into an array format and analyzed simultaneously. The TMA technology is a fast, cost-effective, and statistically powerful method that will substantially facilitate translational research. PMID- 14740492 TI - Generation of double gene disruptions in Dictyostelium discoideum using a single antibiotic marker selection. AB - Gene targeting is a powerful molecular genetic technique that has been widely used to understand specific gene function in vivo. This technique allows the ablation of an endogenous gene by recombination between an introduced DNA fragment and the homologous target gene. However, when multiple gene disruptions are needed, the availability of only a limited number of marker genes becomes a complication. Here we describe a new approach to perform double gene disruptions in Dictyostelium discoideum by simultaneous transfection of two gene targeting cassettes followed by performing clonal selection against only one marker gene. The subsequent PCR-based screens of blasticidin-resistant clones revealed the integration of both the selected and the nonselected targeting cassettes at their original respective loci creating complete gene disruptions. For the genes we have tested in these studies (myosin heavy chain kinases B and C), the efficiency of the double gene targeting event is found in the range of 2%-5% of all blasticidin-resistant colonies following the transfection step. This approach for the simultaneous disruptions of multiple genes should prove to be a valuable tool for other laboratories interested in creating multiple gene disruptants in Dictyostelium or other organisms where a limited number of selectable markers are available. PMID- 14740493 TI - Fluorescent DNA hybridization probe preparation using amine modification and reactive dye coupling. AB - Fluorescent nucleic acid hybridization probes traditionally have been generated by enzymatic incorporation of dye-labeled nucleotides, even though incorporation efficiency is low and variable from dye to dye. Alternatively, 5-(3-aminoallyl) 2'-deoxyuridine 5'-triphosphate (aa-dUTP) is enzymatically incorporated to generate amine-modified DNA, which is then chemically labeled with an amine reactive fluorescent dye. We optimized this latter two-step approach for maximal hybridization signal brightness using DNA probes labeled to varying degrees with different fluorescent dyes. Reverse transcriptase and DNA polymerase 1 efficiently incorporated aa-dUTP into DNA, and adjusting the aa-dUTP:dTTP ratio controlled the degree of substitution. With cDNA probes hybridized to dot blots, probes having approximately eight dyes per 100 bases gave the best sensitivity, irrespective of the dye label. alpha-Satellite probes generated by nick translation and hybridized to human chromosome spreads also showed that probes having approximately eight dyes per 100 bases provided the brightest overall signals. These data demonstrate that this labeling method generates highly sensitive DNA probes that are difficult to obtain by conventional direct incorporation approaches. The technique is inherently consistent and versatile by virtue of the efficient incorporation of primary amines and the reliable chemical labeling reaction. PMID- 14740494 TI - Effect of unlabeled helper probes on detection of an RNA target by bead-based sandwich hybridization. AB - Unlabeled helper oligonucleotides assisting a bead-based sandwich hybridization assay were tested for the optimal placement of the capture and detection probes. The target used was a full-length in vitro synthesized mRNA molecule. Helper probes complementary to regions adjacent to the binding site of the 5' end attached capture probe were found much more effective than helper probes targeting positions adjacent to the detection probe binding site. The difference is believed to be caused by a disruption of the RNA secondary structure in the area where the capture probe binds, thereby reducing structural interference from the bead. The use of additional helpers showed an additive effect. Using helpers at both sides of the capture and detection probes showed a 15- to 40-fold increase in hybridization efficiency depending on the target, thereby increasing the sensitivity of the hybridization assays. Using an electrical chip linked to the detection probe for the detection of p-aminophenol, which is produced by alkaline phosphatase, a detection limit of 2 x 10(-13) M mRNA molecules was reached without the use of a nucleic acid amplification step. PMID- 14740495 TI - Estimation of ribosomal RNA transcription rate in situ. AB - Traditionally, the rate of transcription is measured by metabolic labeling (e.g., the run-on assay), which can be carried out only in isolated or cultured cells. It has been difficult if not impossible to assess the rate of transcription of a gene in a specific cell type in situ. We show here that the quantity of 47S precursor ribosomal RNA (pre-rRNA), which correlates positively with the rate of rRNA transcription as measured by the run-on assay, can serve as an indicator for the rate of its transcription. We adopted this method as an in situ hybridization procedure to demonstrate its validity in vivo. The notion of using the quantity of the primary transcript as an indicator of the rate of transcription has the potential application in monitoring the rate of messenger RNA transcription in single cells within a tissue of complex cellular composition. PMID- 14740496 TI - Enriching for HIV-infected cells using anti-gp41 antibodies indirectly conjugated to magnetic microbeads. AB - The isolation of a pure population of human immunodeficiency virus (HIV)-infected cells is highly desirable for evaluating the impact of HIV on cellular gene expression. Given that HIV gp41 transmembrane protein is anchored on the surface of HIV-infected cells, we evaluated the use of pooled anti-gp41 monoclonal antibodies (MAbs) and HIV immunoglobulins (HIV-Igs) indirectly conjugated to magnetic microbeads to positively select for infected cells. We demonstrate that pooled anti-gp41 monoclonal antibodies enriched for H9 cells infected with HIV IIIB by approximately 98%. Peripheral blood mononuclear cells (PBMCs) infected with a primary (HIV strain 302151) or laboratory-adapted (IIIB) strain were enriched by 54%-62%, depending on the initial viral inoculum. Using HIV-Ig in this magnetic positive-selection approach was also highly efficient for enriching for H9 cells infected with IIIB but less efficient for infected PBMCs. Both types of antibodies used in the selection process resulted in > 80 viability of selected HIV-infected cells. Analysis of interleukin 2 (IL-2) mRNA expression using real-time reverse transcription PCR (RT-PCR) of the HIV-enriched population demonstrated a higher level of IL-2 mRNA, by approximately four cycles, and an 8 fold increase in IL-2 expression, as evaluated by intracellular staining and flow cytometric analysis, in comparison to gp41-negative cells. Collectively, these data illustrate that antibodies targeting gp41 can be used to enrich for HIV positive populations. This represents a novel approach for studying the impact of HIV on infected cells and on bystander/uninfected cells. PMID- 14740497 TI - Nonelectrophoretic method for high-throughput HLA-DRB1 group genotyping. PMID- 14740498 TI - High efficiency transformation by electroporation of Pichia pastoris pretreated with lithium acetate and dithiothreitol. AB - Transformation efficiencies for Pichia pastoris are usually several orders of magnitude below those for other yeast. We report here that pretreatment of P. pastoris with 0.1 M lithium acetate (LiAc) and 10 mM dithiothreitol (DTT) before electroporation increased transformation efficiency approximately 150-fold. DTT alone enhanced the transformation efficiency up to 20-fold, but LiAc alone had little effect. Cultures grown to 1.15-2.6 A at 600 nm had higher transformation efficiencies than younger or older cultures. A cell concentration of 10(10)/mL gave the highest efficiencies. Digestion of pPIC9K within the AOX1 gene with Sacl gave efficiencies approximately 30 times higher than digestion in other genes with other enzymes. Given the optimization of these factors, the highest transformation efficiency was obtained with instrument settings of 1.5 kV, 25 microF, and 186 omega. The transformation efficiency at optimal conditions reached 4 x 10(6) transformants/microgram DNA with pPIC9K. A maximum of 2.6 x 10(5) transformants was produced when 1 microgram of pPIC9K DNA was used. PMID- 14740499 TI - Detection of extremely rare alleles by bidirectional pyrophosphorolysis-activated polymerization allele-specific amplification (Bi-PAP-A): measurement of mutation load in mammalian tissues. AB - Pyrophosphorolysis-activated polymerization (PAP) was developed to detect extremely rare mutations in complex genomes. In theory, PAP can detect a copy of a single base mutation present in 3 x 10(11) copies of the wild-type allele. In practice, the selectivity of detection is limited by a bypass reaction involving a polymerase extension error from the unblocked oligonucleotide annealed to the opposing strand. Bidirectional PAP allele-specific amplification (Bi-PAP-A) is a novel method that uses two opposing 3'-terminal blocked pyrophosphorolysis activatable oligonucleotides (P*s) with one nucleotide overlap at their 3' termini. This eliminates the problematic bypass reaction. The selectivity of Bi PAP-A was examined using lambda phage DNA as a model system. Bi-PAP-A selectively detected two copies of a rare mutated allele in the presence of at least 2 x 10(9) copies of the wild-type lambda phage DNA. Bi-PAP-A was then applied to direct detection of spontaneous somatic mutations in the mouse genome at a frequency as low as 3 x 10(-9). A 370-fold variation in the frequency of a specific somatic mutation among different mouse samples was found, suggesting clonal expansion of mutation occurring during early development and a hyper Poisson variance. Bi-PAP-A is a rapid, general, and automatable method for the detection of rare mutations. PMID- 14740500 TI - Use of complementary and alternative medicine in Israel: 2000 vs. 1993. AB - BACKGROUND: Complementary and alternative medical care has gained increasing popularity in western societies in recent years. OBJECTIVES: To provide a cross sectional and temporal (2000 vs. 1993) analysis of the use of complementary and alternative medicine in Israel. METHODS: The subjects studied represented the Israeli Jewish urban population aged 45-75 years. Full sit-down interviews were conducted with 2,003 respondents in 1993 and 2,505 respondents in 2000. RESULTS: For 1993, 6% of the population reported on consultations with CAM providers during the previous year. For 2000, that proportion increased to 10%. Being a woman, having higher education, enjoying better economic status, being younger, living in a big city, and being dissatisfied with specialists' care were all positively related to the use of non-conventional medicine, particularly in 2000. In both years, more than 50% of the consultations were with acupuncturists and homeopaths. However, chiropractors have doubled their market shares, and lower back pain became the leading problem for which care was sought. The main reason for consulting CAM was a reluctance to use too many drugs or to undergo an invasive procedure. However, a significant proportion of the users continue to use conventional medicine concurrently. Seventy-five percent in 2000 and 60% in 1993 reported that the treatment helped. CONCLUSIONS: Between 1993 and 2000, CAM in Israel changed from an infant industry into a mainstream medical commodity, reflected in both prevalence and different patterns of consumption. PMID- 14740502 TI - The relative merits of Doppler sonography in the evaluation of patients with clinically and scintigraphically suspected testicular torsion. AB - BACKGROUND: Since the early 1970s testicular scintigraphy has been used to diagnose the cause of acute scrotal pain. The advent of Doppler sonography further enhances diagnosis by providing simultaneous real-time scrotal imaging with superimposed testicular blood flow information. OBJECTIVES: To assess the diagnostic value of Doppler sonography in patients with acute scrotal pain and scintigraphic findings suggestive of testicular torsion. METHODS: Seventy-five patients with acute scrotal pain underwent testicular scintigraphy and Doppler sonography. All patients who had scintigraphic findings suggestive of testicular torsion were included in the study and their files were retrospectively reviewed. RESULTS: Twenty-seven patients had scintigraphic findings suggestive of testicular torsion. Radionuclide scintigraphy accurately detected all cases of testicular torsion. However, abscess, hematoma, hydrocele and other conditions simulated testicular torsion on scintigraphy, lowering the test specificity. These pathologies were clarified by Doppler sonography that was 95% specific and 86% sensitive for testicular torsion. CONCLUSIONS: Doppler sonography should be used as the first-line modality in the evaluation of patients with suspected testicular torsion. Scintigraphy should be performed only in certain settings of equivocal sonographic findings to prevent false negative sonographic diagnosis. PMID- 14740501 TI - Positive tissue transglutaminase antibodies with negative endomysial antibodies: low rate of celiac disease. AB - BACKGROUND: Screening for celiac disease is based on the sequential evaluation of serologic tests and intestinal biopsy; an optimal screening protocol is still under investigation. The screening policy of one of the main healthcare providers in Israel (Maccabi) consists of measuring total immunoglobulin A and tissue transglutaminase IgA antibodies and confirming positive results by endomysial antibodies. For IgA-deficient patients antigliadin IgG is measured. OBJECTIVES: To evaluate the use of tTGA as a first-level screening test in patients suspected of having celiac disease METHODS: The results of tTGA and EMA tests over a 3 month period were obtained from the laboratory computer. Letters were sent to the referring physicians of patients with positive tests, requesting clinical information and small intestinal biopsy results. tTGA was performed using an anti guinea pig tTG-IgA enzyme-linked immunosorbent assay kit. RESULTS: Overall, 2,505 tTGA tests were performed: 216 (8.6%) were tTGA-positive of which 162 (75%) were EMA-negative (group 1) and 54 (25%) EMA-positive (group 2.) Clinical information was obtained for 91 patients in group 1 and 32 in group 2. Small intestinal biopsy was performed in 33 (36%) and 27 patients (84%) in groups 1 and 2, respectively. Celiac disease was diagnosed in 4 biopsies (12%) in group 1 and 23 (85%) in group 2 (P < 0.0001). The positive predictive value was 45% for tTGA and 85% for EMA. CONCLUSIONS: Symptomatic patients with positive tTGA and negative EMA have a low rate of celiac disease compared to those who are tTGA-positive and EMA-positive. Confirmation with EMA is advised when tTGA is performed as a first level screening for suspected celiac disease. PMID- 14740503 TI - Anemia as a risk factor for ischemic heart disease. AB - BACKGROUND: Anemia is a known risk factor for ischemic heart disease. Based on knowledge of the physiologic role of oxygen delivery to the myocardium, anemia may be a cause of more severe cardiovascular diseases or a marker of other processes occurring in the body that induce more severe disease. OBJECTIVES: To investigate the relationship between anemia and the clinical picture of IHD, including manifestations, severity and complications. METHODS: The population studied comprised 417 similarly aged patients with IHD and anemia. The patients were categorized into subgroups of IHD according to disease severity: namely, angina pectoris, acute ischemia, acute myocardial infarction, congestive heart failure or cardiac arrhythmias. Two populations served as control groups: patients with anemia but no IHD (C-I) and patients with IHD without anemia (C II). A standard anemia workup was conducted in all patients with IHD and anemia and a correlation was made between the hematologic parameters and the manifestations and complications of IHD. RESULTS: The common presenting symptom was chest pain in the study group and in C-II (94% and 86% respectively) and weakness (90%) in C-I. Patients with IHD and anemia tended to suffer from a more advanced degree of IHD (80%) compared to patients with IHD alone who had milder disease (46%). Hematologic values including hemoglobin, mean cell volume, serum iron and total iron-binding capacity correlated inversely with disease severity among anemic patients with IHD. There were significant differences between the study group and C-II regarding CHF (31% and 18% respectively) and arrhythmias (41% and 16% respectively). The mortality rate was higher in patients with IHD and anemia than in patients with IHD alone (13% and 4% respectively). CONCLUSIONS: Anemia is a significant risk factor in IHD. It correlates with advanced IHD, CHF, rhythm disturbance and higher mortality rate. An aggressive therapeutic and preventive approach might improve the outcome of this disease. PMID- 14740504 TI - Consumption of opioids in a hospital setting--what can we learn from a 10 year follow-up? AB - BACKGROUND: The World Health Organization considers a country's morphine consumption to be an important indicator of progress in pain relief. Despite the strong consensus favoring the use of opioids in many types of pain, limited data are available for gauging the trends in opioid usage in specific medical institutions, such as hospitals. OBJECTIVES: To assess the possibility that monitoring opioid consumption can shed light on directions and trends in the treatment of pain in a hospital setting. METHODS: Data on opioid consumption, number of inpatient days and number of operations performed each year during the period 1990-1999 were obtained from records kept in the hospital's pharmacy and archives. RESULTS: During that decade the overall opioid consumption in the hospital increased from the equivalent of 3.7 mg of oral morphine per inpatient day to 7.3 mg, and from 56 mg per surgical procedure to 100 mg. In 1990, injected opioids accounted for 93% of the overall consumption, whereas in 1999 they accounted for only 44%. Yet, the proportion of injected meperidine to injected morphine increased only from 43% to 51%. CONCLUSIONS: These results suggest that the ongoing monitoring of opioid consumption can highlight trends and directions and possibly emphasize strengths and weaknesses in the treatment of pain in hospitals. PMID- 14740505 TI - Risk of transmission of leptospirosis from infected cattle to dairy workers in southern Israel. AB - BACKGROUND: Leptospirosis is a zoonotic disease that occurs worldwide, found predominantly in agricultural workers, port workers and dairy workers. OBJECTIVE: To investigate the risk of disease transmission to dairy workers following an outbreak in 1999 of Leptospirosis hardjo in the dairy herds of two kibbutzim in southern Israel. METHODS: A seroepidemiologic survey of all the dairy workers from these two kibbutzim was conducted, including individual interview and examination. Data were collected on the presence of clinical symptoms of leptospirosis during the previous month. One month later the medical personnel on the two kibbutzim were contacted in order to determine if any worker had subsequently developed clinical signs or symptoms of leptospirosis. All dairy workers had blood drawn for serology. Those workers whose initial serology had been borderline for leptospirosis had a repeated serology test between 2 and 4 weeks later. Doxycycline was given prophylactically to all dairy workers on one kibbutz only. RESULTS: Either with or without chemoprophylaxis, no dairy workers exposed to herds infected with Leptospira hardjo showed evidence of seroconversion or disease. This indicated a low risk of transmission of this serovar from cows to dairy workers. CONCLUSION: Since human illness with leptospirae can cause illness associated with significant morbidity, we recommend that physicians make an informed decision regarding doxycycline prophylaxsis for dairy workers exposed to cattle herds infected with Leptospira hardjo. PMID- 14740506 TI - Therapeutic contact lens as the primary treatment for traumatic corneal erosions. AB - BACKGROUND: Corneal erosions, a common and very painful ailment, are traditionally treated with pressure patches and antibiotic ointment but the healing is slow. OBJECTIVES: To report our experience with the use of therapeutic contact lenses for the primary treatment of traumatic corneal erosions. METHODS: During the last 5 years in a single community clinic 65 consecutive patients with traumatic corneal erosions were treated with a corneal contact lens and antibiotic drops as a routine measure. The charts were reviewed for outcome, side effects and complications. RESULTS: Healing of the corneal erosions occurred within 1 to 3 days in all patients, with minimal or no pain. No corneal infection occurred. One patient had a recurrence that was successfully treated by lens placement. CONCLUSIONS: The therapeutic contact lens with antibiotic drops is a safe and effective method to treat traumatic corneal erosions, and patients can immediately resume their regular activities. PMID- 14740507 TI - Correlation between iron status and genetic hemochromatosis (codon C282Y) in a large German population. AB - BACKGROUND: Genetic hemochromatosis leads to iron overload in many tissues and may lead to liver cirrhosis and hepatocellular carcinoma. Early diagnosis and therapy are crucial. Since 80-100% of hemochromatosis patients of European origin are homozygous for a cysteine to tyrosine exchange in the HFE gene at codon 282, genetic screening might be useful. Representative population studies are needed to evaluate the phenotype of people heterozygous and homozygous for the C282Y mutation. OBJECTIVE: To determine the correlation between parameters of iron metabolism and the hemochromatosis genotype in a large population-based study. METHODS: A representative population-based survey, the Diabetomobil study, analyzed 5,083 German probands. Serum transferrin saturation and ferritin levels were determined, and the C282Y mutation of the HFE gene was analyzed by restriction fragment length polymorphism-polymerase chain reaction analysis. RESULTS: Nine of 373 probands with a transferrin saturation > 55% (2.4%) and none of 264 randomly selected probands with a transferrin saturation < or = 55% (0%) were homozygous for the C282Y mutation. Three of the nine homozygous probands had ferritin values less than 250 micrograms/L. The frequency of the heterozygous genotype was 8.8%, and the percentage of heterozygous probands increased with increasing levels of transferrin saturation. CONCLUSION: We propose a population screening strategy with an initial transferrin saturation test, followed by genotyping for the C282Y mutation if the transferrin saturation is above 55%, regardless of the ferritin level. Heterozygous individuals with higher transferrin saturation values may be protected against iron loss but may also be more susceptible for certain liver diseases, depending on the simultaneous prevalence of other diseases. PMID- 14740508 TI - Palliative treatment for advanced or metastatic osteosarcoma. PMID- 14740509 TI - Cord blood biology and transplantation. AB - In recent years, umbilical cord blood has emerged as an alternative source of hematopoietic progenitors (CD34+) for allogeneic stem cell transplantation, mainly in patients who lack an human leukocyte antigen-matched marrow donor. Since 1998, about 2,500 patients have received UCB transplants for a variety of malignant and non-malignant diseases. The vast majority of recipients were children with an average weight of 20 kg, however more than 500 UCB transplantations have already been performed in adults. The "naive" nature of UCB lymphocytes may explain the lower incidence and severity of graft versus host disease encountered in UCBT compared to the allogeneic transplant setting. Furthermore, UCB is rich in primitive CD16-CD56++ natural killer cells, which possess significant proliferative and cytotoxic capacities and can be expanded using interleukin-12 or 15, so as to mount a substantial graft versus leukemia effect. The major disadvantage of UCB is the low yield of stem cells, resulting in higher graft failure rates and slower time to engraftment compared to bone marrow transplantation. A rational approach thus involves ex vivo expansion of UCB-derived hematopoietic precursors. PMID- 14740510 TI - "Knowledge stems from disagreements". PMID- 14740511 TI - Clinical and laboratory assays in the diagnosis of cutaneous adverse drug reactions. PMID- 14740512 TI - The cerebral hyperperfusion syndrome: what every family doctor should know. PMID- 14740513 TI - The hyperperfusion syndrome: an under-recognized complication of carotid endarterectomy. PMID- 14740514 TI - Idiopathic liver involvement in Turner syndrome. PMID- 14740515 TI - Primary greater omental torsion: CT diagnosis in an elderly woman. PMID- 14740516 TI - In vitro interferon-gamma release test in the diagnosis of drug-induced erythema nodosum. PMID- 14740517 TI - Splenic rupture after colonoscopy. PMID- 14740518 TI - Computed tomography: not an ophthalmic polygraph. PMID- 14740519 TI - Secondary hypertrophic osteoarthropathy (HOA) mimicking primary HOA (pachydermoperiostitis or Touraine-Solente-Gole) syndrome. PMID- 14740520 TI - Primary stability of threaded cups in THR--an experimental study. AB - Four threaded cups were tested up to their lever-out moments, torque-in moments and their resistance to failure. A was a parabolic-shaped, B was a spherical, C was a spherical-shaped too, and D was a conical shaped cup. Cup A and D represent cups which have proven themselves in clinical applications, but not cup B. The threads were determined and showed different constructive features. The cups were torqued into precise cavities in PVC foam cubes, after that they were levered out in a testing machine. The lever-out moments of all the cups showed significant differences; the results were: A: 78.4 Nm, B: 88.7 Nm, C: 117.5 Nm, D; 136.6 Nm. In the case of the torque-in moments there were no significant differences between A and B, neither between C and D. The differences in stiffness between B and C were not significant, but they were between the others. The primary stability against lever-out and the torque-in moment of threaded cups for artificial hip replacement can be basically influenced by different constructive features. Hence lever-out moment and torque-in moment should be understood and tested as independent variables. PMID- 14740521 TI - [Useability of an image based nagivation system in reconstruction of leg alignment in total knee arthroplasty - results of a propsective study]. AB - Accurate leg alignment is one important factor for long term survival in total knee arthroplasty (TKA). The aim of this study was to determine the accuracy of a CT-based navigation system in restoration of the leg axis. 80 TKA (P.F.C.-Sigma- DePuy) were implanted either using a CT-based navigation-system or the conventional technique. There were no significant differences between the groups according to the preoperative leg deformity. Accuracy of implantation was determined on postoperative long-leg coronal and lateral x-rays. A postoperative leg axis between 3 degrees varus and 3 degrees valgus was obtained in 35/40 patients (88%) in the computer assisted and 28/40 patients (70%) in the conventional operated group. A higher precision in the sagittal plane was seen for the tibial (computer assisted: 3.9 +/- 2.2 vs. conventional TKA 1.3 +/- 5.8) and the femoral component (computer assisted: 3.5 +/- 2.3 vs. conventional TKA 6.9 +/- 3.6). The use of the CT-based VectorVision-System allows a significant improvement in the accuracy of implantation in TKA. The CT-based module has the advantage of a precise preoperative planning and a useful documentation and control tool for each important step. On the other hand there are additional costs and time consuming logistics. PMID- 14740522 TI - [Radiological evaluation of polyethyelene wear of cementless total hip endoprosthesis]. AB - During the last years increasingly cementless hip endoprostheses have been implanted. Radiological wear measurement of cemented hip endoprostheses for the material couples polyethylene cup-ceramic- or metal head has been established in the literature. However, for cups encased by metal (screwing or pressfit cups), this method of measurement is not applicable. Therefore, a method has been developed to measure wear on radiographs on cementless spherical implants. The data were compared to those, obtained from conventional wear measurements on cemented hip cups. The results indicate that both techniques generate comparable results, thus validating the new technique as being suited for cementless cup implants. PMID- 14740523 TI - [Biomechanicsl evaluation of a stand-alone interbody fusion cage based on porous TiO2/glass-ceramic on the human cervical spine]. AB - Recently, there has been a rapid increase in the use of cervical spine interbody fusion cages, differing in design and biomaterial used, in competition to autologous iliac bone graft and bone cement (PMMA). Limited biomechanical differences in primary stability, as well as advantages and disadvantages of each cage or material have been investigated in studies, using an in vitro human cervical spine model. 20 human cervical spine specimens were tested after fusion with either a cubical stand-alone interbody fusion cage manufactured from a new porous TiO2/glass composite (Ecopore) or PMMA after discectomy. Non-destructive biomechanical testing was performed, including flexion/extension and lateral bending using a spine testing apparatus. Three-dimensional segmental range of motion (ROM) was evaluated using an ultrasound measurement system. ROM increased more in flexion/extension and lateral bending after PMMA fusion (26.5%/36.1%), then after implantation of the Ecopore-cage (8.1%/7.8%). In this first biomechanical in vitro examination of a new porous ceramic bone replacement material a) the feasibility and reproducibility of biomechanical cadaveric cervical examination and its applicability was demonstrated, b) the stability of the ceramic cage as a stand alone interbody cage was confirmed in vitro, and c) basic information and knowledge for our intended biomechanical and histological in vivo testing, after implantation of Ecopore in cervical sheep spines, were obtained. PMID- 14740524 TI - Anterior vertebral body screw pullout testing with the hollow modular anchorage system--a comparative in vitro study. AB - Pullout of implants at the proximal and distal ends of multilevel constructs represents a common spinal surgery problem. One goal concerning the development of new spinal implants is to achieve stable fixation together with the least invasive approach to the spinal column. This biomechanical study measures the influence of different modes of implantation and different screw designs, including a new monocortical system, on the maximum pullout strength of screws inserted ventrolaterally into calf vertebrae. The force pullout of eight different groups were tested and compared. Included were three bicortical used single screws (USS, Zielke-VDS, single KASS). To further increase pullout strength either a second screw (KASS) or a pullout-resistant nut can be added (USS with pullout nut). A completely new concept of anchorage represents the Hollow Modular Anchorage System (MACS-HMA). This hollow titanium implant has an increased outside diameter and is designed for monocortical use. Additionally two screw systems suitable for bicortical use were tested in monocortical mode of anchorage (USS, single KASS). We selected seven vertebrae equal in mean size and bone mineral density for each of the eight groups. The vertebral body and implant were connected to both ends of a servohydraulic testing machine. Displacement controlled distraction was applied until failure at the metal-bone-interface occurred. The maximum axial pullout force was recorded. Mean BMD was 312 +/- 55 mg CaHA/ml in cancellous bone and 498 +/- 98 mg CaHA/ml in cortical bone. The highest resistance to pullout found, measured 4.2 kN (KASS) and 4.0 kN (USS with pullout nut). The mean pullout strength of Zielke-VDS was 2.1 kN, of single KASS 2.5 kN, of MACS-HMA 2.6 kN and of USS 3.2 kN. There was no statistically significant difference (t-test, p > 0.05) between bicortical screws and the new monocortical implant. For the strongest fixation at the proximal or distal end of long spinal constructs the addition of a second screw or a pullout-resistant nut behind the opposite cortex offers even stronger fixation. PMID- 14740525 TI - Instrumented forceps for measuring tensile forces in the rod of the VDS implant during correction of scoliosis. AB - Ventral derotation spondylodesis (VDS) is the standard in ventral scoliosis surgery. Especially in the thoracic spine, there are no alternatives to VDS with compression and derotation as its correction forces. However, pull-out of the end vertebra screw during correction of scoliosis with the VDS implant is a common complication involving particularly the cranial end-vertebra screw in the thoracic region. This complication requires an extension of the fusion length or reduces at least the outcome of the correction. There are no in vivo data on correction forces in ventral scoliosis surgery. Thus the correction depends on the skill and experience of the surgeon. An instrumented forceps developed and built to measure forces in the longitudinal rod allows axial tensile forces to be determined in the longitudinal rod during surgery. The instrumented forceps has the advantage of reducing the risk of screw pull-out. Furthermore, viscoelastic behavior of the spine can be measured during ventral correction. In addition, knowledge of the correction forces improves our biomechanical understanding of the spine, especially during correction of scoliosis. Intraoperative force measurement is in no way detrimental to the patient. PMID- 14740526 TI - [Malignant diseases in children--challenges and perspectives]. PMID- 14740527 TI - [Diagnostic significance of synovial biopsy in patients with Reiter's syndrome]. AB - INTRODUCTION: Arthritis in Reiter's syndrome (RS) is a reactive synovitis associated with a localized infection of the urogenital or gastrointestinal tract with a genetic predisposition. The pathogenetic mechanisms for synovitis in RS are still unknown. Our aim was to examine some of the pathogenetic mechanisms in Reiter's syndrome looking for morphologic changes, immunoprotein deposits and microorganism antigens in synovial biopsies and to determine whether synovial biopsy is useful in diagnosis of RS. MATERIAL AND METHODS: Thirty patients with urogenital form of RS were examined within a four-year period. Table 1 illustrates laboratory findings in our patients. We performed synovial biopsies looking for histopathological changes, deposits of immunoproteins and microorganism antigens. Analysis of synovial biopsy specimens was performed using light and immunofluorescence microscopy and fluorescein-labelled monoclonal antibodies to Chlamydia trachomatis. RESULTS: Histopathological examination of synovial membrane revealed marked proliferation of the synovial lining cells (SLC) with less or more abundant papillary projections, hypertrophic and edematous tissue with marked vascularisation in 28 (93.3%) cases. Fibrinoid necrosis foci were seen on the surface of synovial tissue. Chronic inflammatory cells (CIC) were diffusely distributed. Edema of the vessel walls, swollen endothelial cells, fibrinoid necrosis in vessel walls as well as multilaminated basement membranes were observed. All histopathologic changes are presented in Table 2. Immunofluorescent techniques in 12 out of 30 (40%) synovial membranes showed immunoglobulin deposits: IgG and IgA deposits were found in vessel walls in 7 cases each and IgM in 10 biopsy specimens. C3 was present perivascularly or within the vessel wall in 4 (13.3%) cases. Sections treated using fluorescein conjugated antibody revealed Chlamydia in the synovial tissue in 2 patients. CONCLUSION: Biopsy specimens with previously described changes in patients with suspected Reiter's syndrome can be useful to confirm the diagnosis. According to our experience, multiple biopsies of abnormal synovia are recommended in these patients. PMID- 14740528 TI - [Effect of smoking on resorption of residual alveolar ridges in complete denture wearers]. AB - INTRODUCTION: Based on literature data it is obvious that there is a connection between smoking and periodontal diseases. Alveolar bone loss increases with smoking. Tobacco smoking affects the proportion of subgingival bacterial flora by influencing oxidoreduction potential of dental plaque and thus making conditions for development of anaerobic bacteria. According to some researchers, smoking affects the mineral component of bone tissue. Orthopantomograms show higher level of alveolar bone loss in smokers than in nonsmokers with the same level of oral hygiene. The aim of this study was to establish if smoking affects alveolar bone loss in complete denture wearers. MATERIAL AND METHODS: Our clinical investigation included 60 patients of both sexes (30 smokers and 30 nonsmokers) all complete dentures wearers. All patients met study criteria: jaw relation and smokers who smoke over 20 cigarettes per day. All subjects were interviewed, and after that orthopantomograms were made. They were used to calculate the degree of alveolar bone loss. RESULTS: The examined subjects were approximately of the same age. Mean age of smokers was 59.9 and nonsmokers 61.8. It was established that differences regarding resorption in men were not significant. The degree of resorption in women smokers and women non-smokers was different, but differences were not significant. DISCUSSION: It has been proven that the number of cigarettes smoked per day is very important. It is considered that the risk of oral epithelial dysplasia increases when smoking more than 20 cigarettes per day. Considering our results regarding resorption of edentulous alveolar ridge in smokers and nonsmokers, we concluded that there were no significant differences. There are opinions in literature that smoking is not an etiological factor in resorption, but there are some opinions that smoking is connected with the degree of resorption in periodontium. The analyses of resorptive changes in edentuolous smokers were done only around implants and it was estimated that smoking has more influence than other clinical risk factors. CONCLUSION: On the bases of our research we may conclude that smoking does not directly affect the degree of resorption of edentulous alveolar ridge with complete denture wearers. PMID- 14740529 TI - [Low power laser biostimulation in the treatment of bronchial asthma]. AB - INTRODUCTION: Modern concept of acupuncture is based on the fact there are designated locations on the surface of human body, which are related to integrative systems of an organism by means of sensory nerves, correlating and synchronizing organ functioning, depending on external and internal conditions, by means of nervous and neurohumoral regulation of metabolic and regenerative processes, including also mobilisation of immunological, protective and antistress reactions. Apart from standard needle acupuncture, other methods of stimulating acupuncture points are also applied. Due to invention of low power lasers, irradiation laser acupuncture has been introduced into routine medical practice, characterised by painless and aseptic technique and outstanding clinical results. MATERIAL AND METHODS: The investigation was aimed at defining therapeutic effects of low power laser irradiation by stimulating acupuncture points or local treatment of asthma. A prospective analysis included 50 patients treated at the Institute of Pulmonary Diseases in Sremska Kamenica during 2000, 2001 and 2002. Together with conservative treatment of present disease, these patients were treated with laser stimulation of acupuncture points in duration of ten days. During treatment changes of functional respiratory parameters were recorded. Results were compared with those in the control group. The control group consisted of the same number of patients and differed from the examination group only by not using laser stimulation. RESULTS: Patients with bronchial asthma presented with significant improvement (p < 0000,5) of all estimated lung function parameters just 30 minutes after laser stimulation. Improvements achieved on the third and the tenth day of treatment were significantly higher (p < 0.001 to p < 0.00005) in the examination group in comparison with the control group. Further investigation confirmed that improvement of measured lung function parameters was significantly higher in younger patients, in patients whose disease lasted shorter, as well as in women. Patients with asthma, who were treated every three months for a one year period, presented with significantly lower frequency and intensity of attacks. DISCUSSION: The mechanism of laser stimulation activity in treatment of bronchial asthma is explained in detail, correlating our results to those obtained by other authors. CONCLUSIONS: A ten day course of low-power laser stimulation of acupuncture points in patients with bronchial asthma improves both the lung function and gas exchange parameters. Positive effects of laser treatment in patients with bronchial asthma are achieved in a short time and they last long, for several weeks, even months. Successive laser stimulation in asthmatics prolongs periods of remission and decreases the severity of asthmatic attacks. Better positive effects of laser stimulation are achieved in younger asthmatics, in those with shorter disease history and in female patients. There is a negative correlation between effects of laser stimulation and patients' age and disease history. However, these characteristics do not affect response rate and speed but positive laser stimulation effects are achieved in a shorter period in female asthmatics. PMID- 14740530 TI - Optimal therapy of epilepsy by measuring serum concentration of antiepileptic drugs with least adverse effects. AB - The study comprised 150 epileptic patients treated at the Institute of Neurology of the Clinical Centre Novi Sad. The optimal therapy with least adverse effects and seizures was achieved in patients in whom measurement of serum concentration of antiepileptic drugs was performed. Patients were divided into five groups with respect to the therapy they received: I--carbamazepane; II--valproic acid; III- polytherapy with phenobarbitone and diphenylhydantoin; IV--phenobarbitone and valproic acid; and V--phenobarbitone, valproic acid and carbamazepine. No adverse effects were recorded in over 60% of patients on monotherapy, 35% of patients who received two anticonvulsants, and 30% of patients who received three anticonvulsants. Significant correlation between drug dosage and blood drug concentration (r > 0.5) was found in polytherapy with phenobarbitone, carbamazepine and valproic acid (r = 0.66); and phenobarbitone and diphenylhydantoin (r = 0.53). PMID- 14740532 TI - [Postoperative nausea and vomiting]. AB - INTRODUCTION: Postoperative nausea and vomiting (PONV) and pain are the most common complaints following anesthesia and surgery, and due to negative emotional impact on patients, they cause prolonged postoperative recovery. The incidence of PONV is 20-30% during the first 24 h after anesthesia. Both peripheral and central mechanisms are involved in control of emesis. FACTORS: Many factors associated with anesthesia and surgery may affect PONV: patient's age and sex, history of PONV after pervious anesthesia, administration of antiemetics prior to operation, type and duration of operation, type of premedication, induction agent, maintenance agent, reversal of muscle relaxation, treatment of postoperative pain and movement of patients. ANTAGONISTS OF 5-HYDROXYTRYPTAMINE-3 (5-HT3) RECEPTORS: Ondansetron is a competitive serotonin type 3 receptor antagonist important in prevention of PONV. However, if 5-HT 3 receptor antagonists are effective against nausea and vomiting after a variety of anesthetic and surgical procedures, then at last we may have the key to the mechanism of PONV. Prophylaxis with a combination of antiemetic drugs is more effective in prevention of PONV. PMID- 14740531 TI - [Treatment of chronic hepatitis C with PEG-interferon]. AB - INTRODUCTION: Since the discovery of the hepatitis C virus, the etiology of chronic liver diseases has been revealed in great number of patients. However, the treatment of hepatitis C viral infection still hasn't been completely resolved. Antiviral and immunomodulatory effects of interferon, and antiviral effect on the nucleoside analogs were efficient only in small number of patients. Discovery of pegylated interferon brings progress in therapeutic success rates. MATERIAL AND METHODS: Combined therapy with peginterferon alfa-2a (Pegasys) 180 mg once a week plus Ribavirin 800 mg a day during a 24-week period was conducted in 20 patients (13 were previously treated with standard antiviral therapy). The aim of this study was to determine the safety and the efficacy of therapy in our patients. RESULTS AND DISCUSSION: Analysis of safety of the combined therapy was conduced in all 20 patients, and analysis of efficiency in 18 patients. Efficacy of the combined therapy was assessed regarding to biochemical response (normalization of aminotransferase activity at the end of therapy and at the end of 6-month follow-up) and virologic response (disappearance of RNA HCV in serum at the end of 6-month follow-up). 30% of treated patients experienced no troubles during treatment. Influenza-like symptoms, weight loss, depression, hair loss and reaction at the site of injection were mild and did not exclude patients from their usual activities in family, society and work place. Neutropenia, thrombocytopenia and anemia as well as elevated aminotransferase activity demanded periodical dose modification in 20% of patients. Unexpected unwanted effect emerged in one patient after cessation of therapy (pulmonary sarcoidosis). Good effects of combined therapy at the end of follow-up period showed biochemical and virologic response in 66% of patients. CONCLUSION: Combined therapy with Pegasys 180 mg/week and Ribavirin 80 mg/day is safe and well tolerated. Sustained biochemical and virologic response was achieved in 66% of patients. PMID- 14740533 TI - [Children without parental care as a vulnerable population group]. AB - INTRODUCTION: Health protection and health promotion are the primary goals of modern medicine. Since children's health is the cornerstone of adult's health, it must be in the center of all social and health care strategies. VULNERABILITY CONCEPT: Vulnerability means exposure to harmful influences, risks or stress, which increases disease probability. The most vulnerable are categories that are exposed to influence of many harmful factors, which have minimal chances for survival and lowest quality of life. VULNERABILITY OF CHILDREN WITHOUT PARENTAL CARE: Vulnerability of children without parental care is caused by lack and/or inadequate family environment. These children are usually emotionally unstable; they frequently develop conduct disorders and have low self-esteem. As adolescents, these children have tendency to risky behavior, which greatly decreases their health potentials. HEALTH STATUS AND QUALITY OF LIFE: All three components of health are endangered with children without parental care. These children present with physical, psycho-motoric and intellectual impairments. There are no studies about quality of life regarding these children, but we can assume that their quality of life is lower than in children who experience protective family environment, since quality of psychosocial factors and family environment are very important predictors of quality of life. CONCLUSION: Children without parental care are an extremely vulnerable category, because they are subjected to various risk factors. Therefore, in order to improve health potentials and quality of life, special measures are required in health care, psychological care and social welfare. PMID- 14740534 TI - [Ketamine: the past 30 years and its future]. AB - INTRODUCTION: There are two isomers of ketamine: S(+) ketamine and R(-) ketamine. Effects of this drug are mediated by N-methyl-D-aspartate (NMDA), opioid and muscarinic receptors. PHARMACOKINETICS: Due to its high lipid solubility and low protein binding, ketamine is extensively distributed in the body. Ketamine metabolism is mediated by hepatic microsomal enzymes. EFFECTS ON BODY SYSTEMS: Ketamine causes increased intracranial pressure, bronchodilation and stimulation of cardiovascular system. CLINICAL USE: ANESTHESIA: It is used for premedication, sedation, induction and maintenance of general anesthesia. Ketamine is an ideal anesthetic agent for trauma victims, patients with hypovolemic and septic shock, patients with pulmonary diseases. ANALGESIA: Even subanesthetic doses of ketamine have analgesic effects, so ketamine is also recommended for postoperative analgesia. Recent results show that ketamine is not always effective as an analgesic in treatment of chronic pain and that some side effects limit its use in patients with chronic pain. INTENSIVE CARE: Combination of ketamine and midazolam can be very useful and safe for sedation and pain relief in intensive care patients, especially during ventilator management. Ketamine may be the anesthetic of choice for painful procedures in intensive care units. PMID- 14740535 TI - [Sex hormones and female sexuality]. AB - INTRODUCTION: In contrast to animal species in which linear relationships exist between hormonal status and sexual behaviour sexuality in human population is not determined so simply by the level of sexual steroids. The article analyses female sexuality in the light of hormonal status. ADMINISTRATION OF SEXUAL STEROIDS DURING PREGNANCY AND SEXUAL DIFFERENTIATION: High doses of gestagens, especially those with high androgen activity, widely used against miscarriages may lead to tomboys, but without differences in sexual orientation. However, it has been observed that the frequency of bisexual and lesbian women is higher in women with congenital adrenogenital syndrome. HORMONES SEXUAL DESIRE AND SEXUALITY DURING MENSTRUAL CYCLE: It has been established that sexual desire, autoeroticism and sexual fantasies in women depend on androgen levels. There are a lot of reports claiming that sexual desire varies during the menstrual cycle. HORMONAL CONTRACEPTION AND SEXUALITY: Most patients using birth control pills present with decreased libido. But, there are reports that progestagens with antiandrogenic effect in contraceptive pills do not affect sexual desire. HORMONAL CHANGES IN PERI- AND POSTMENOPAUSAL PERIOD AND SEXUALITY: Decreased levels of estrogen and testosterone in older women are associated with decreased libido, sensitivity and erotic stimuli. SEXUALITY AND HORMONE REPLACEMENT THERAPY: Hormonal therapy with estrogen is efficient in reference to genital atrophy, but not to sexual desire. Really increased libido is achieved using androgens. Also, therapy with dehydroepiandrosterone (DHEA) and tibolone have positive effects on female libido. CONCLUSION: Effect of sexual steroids on sexual sphere of women is very complex. The association between hormones and sexuality is multidimensional, as several hormones are important in regulation of sexual behaviour. Still, it should be pointed out that sexuality is in the domain of hormonal, emotional motivational and social factors. PMID- 14740536 TI - [Importance of extensive lymphadenectomy in relation to the extent of metastatic lymph node involvement in patients with gastric carcinoma surgery]. AB - INTRODUCTION: Formation of lymphatic metastasis is an important prognostic factor in treatment of gastric carcinoma. In this paper we would like to point to the importance of systematic lymphadenectomy in regard to extent of metastatic lymph node involvement in patients with gastric carcinoma. MATERIAL AND METHODS: This investigation included 114 patients with diagnosis of gastric carcinoma undergoing systematic lymphadenectomy. We analyzed metastatic lymph node involvement considering: 1. tumor localization; 2. histopathologic type; 3. depth of tumor invasion; 4. extent of lymph node resection; 5. stage of disease. RESULTS: In the course of this study, 2100 lymph nodes were extracted by systematic lymphadenectomy (18.42 lymph nodes per patient on average). Out of this number, about 27% of lymph nodes were involved with metastasis, and 60% of positive lymph nodes belonged to the first, 25.7% to the second, 10.2% to the third and 3.8% to the fourth drainaged group. The most frequent localization (over 56%) of tumors was the distal third region of stomach. Histopathologically, most common type of carcinoma was intestinal carcinoma, with metastasis in first and second drainage group. A great number of patients have tumors with infiltration of the serosa. Those patients had metastases in lymph nodes in over 42%. Most patients (58%) belonged to IV stage of disease and presented with highest number of involved metastatic lymph nodes in all drainage groups. DISCUSSION: In our patients high metastatic involvement of all four drainage groups of lymph nodes was established. Up to date standard lymphadenectomy (D1) which has been performed increased the length of survival of these patients. CONCLUSION: By systematic lymphadenectomy we removed a great number of metastatic lymph nodes with beneficial effect on survival of patients with gastric carcinoma. PMID- 14740537 TI - [Suppurative middle ear infection as a complication after tympanostomy tube placement]. AB - INTRODUCTION: Suppurative otitis media after tympanostomy tube placement is the most frequent complication of this surgical intervention. Otorrhea that occurs in the first two weeks following tube placement is called early, late otorrhea occurs at least two weeks following placement. Early otorrhea is usually a result of either an infection that already existed when the tube was placed, or contamination of the external auditory canal during operation. Late otorrhea is mostly a result of upper respiratory tract infection. MATERIAL AND METHODS: Our investigation was performed at the ENT Department, Mother and Child Health Care Institute in Belgrade. The research included children treated for secretory or recurrent otitis media. RESULTS AND DISCUSSION: We have examined 411 children implanted with 796 tympanostomy tubes in the last three years. We investigated changes within two weeks after operation. Suppuration was recorded with 81 children (19.7%). Staphylococcus aureus was established in 33 (40.7%) Pseudomonas aeruginosa in 26 (32%), Haemophilus influenzae in 12 (15%) and Streptococcus pneumoniae in 10 (12.3%) cases. All children were treated with antibiotic ear drops according to the antibiogram for a period of 7 days. Full recovery was achieved after treatment with Ciprofloxacin drops in 67%, Neomycin in 18% and Gentamycin in 9% of cases. In other cases a combination of drops and oral antibiotics was used. CONCLUSION: In cases of suppurative otitis media after implantation of tympanostomy tubes, the secret should be treated with suction and after that antibiotic drops should be applied during 5 to 7 days. If suppuration is persistent, drops should be used with oral antibiotics. PMID- 14740538 TI - [Catheter-related urinary infections at the Clinical Center in Banja Luka]. AB - INTRODUCTION: Catheter-associated urinary tract infections are the most common nosocomial infections of the urinary tract, and among the most common nosocomial infections in general. The major problems of these infections include antibiotic resistance and enormous direct and indirect cost of treatment. MATERIAL AND METHODS: A retrospective study on major causes of infections and antibiotic resistance was conducted at four clinics of the Clinical Center of Banja Luka. An anonymous questionnaire was distributed to nursing staff dealing with urinary catheters in order to get an overview of their clinical performance. RESULTS: The results showed that in 89% of cases (out of 198 patients with developed catheter associated urinary tract infection) infections were caused by gram-negative bacteria, in 7% by gram-positive bacteria and in 4% by candida. The most common bacteria were: Escherichia coli (33.6%), Pseudomonas aeruginosa (14.1%), Proteus mirabilis (13.3%), and Enterobacter (10.5%). Majority of bacteria presented with extremely high resistance (72-100%) to ampicillin, gentamycin and cotrimoxazole, and in some cases a significant resistance to ciproflaxacine, nalidixic acid, ceftriaxone and ceftazidime. The questionnaire showed that nursing staff did not follow guidelines for medical care of patients with urinary catheters. CONCLUSION: It can be concluded that poor hygienic and epidemiological conditions, as well as irrational use of antibiotics contribute to uncontrolled development of urinary tract infections in catheterized patients. PMID- 14740539 TI - Research on psychopathological consequences of refugeeism. AB - The study examined mental and psychopathological consequences of refugeeism and included (109) refugees from refugee camp in Krnjaca. Their reactions were compared with the reactions of (70) somatically injured patients from Orthopedic Hospital of the Clinical Centre in Belgrade and subjects (105) from Belgrade denying any traumatic experience whatsoever. Apart from the questionnaire on socio-demographic features, subjects were asked to provide answers to Post Traumatic Stress Disorder (PTSD-10) scale, Brief Eysenck's Personality Inventory, Family Homogeneity Inventory and Impact of Events Scale. By implementation of appropriate statistical procedures (variance analysis), the significance of differences among certain features within the experimental group of refugees was examined, as well as the difference concerning presence of mental and psychopathological features among all three subgroups. It was determined that within the refugee group, males more significantly and more frequently reacted with PTSD symptoms, as well as with signs of general neuroticism and that married refugees more frequently presented with PTSD symptoms. In comparison to non refugees, refugees more significantly and more frequently present with PTSD symptoms, which is probably caused by PTSD chronicity in this category of subjects. Subjects denying any significant traumatic experience in their lives presented with symptoms of general neuroticism more than the rest of examinees. This is an interesting finding that can be explained by the fact that the same group achieved the highest values on the extraversion scale (using the same Eysenck instrument), or may be correlated to the increase of neurotic reactions in the whole population of Serbia and Montenegro. The above mentioned and other results were compared to the findings of similar researches performed by other authors. PMID- 14740540 TI - [Obesity and hormone function changes in female patients with polycystic ovaries]. AB - INTRODUCTION: Polycystic ovary syndrome is commonly associated with hyperandrogenism and anovulation. The aim of this study was to investigate the impact of obesity on hormonal status in patients with polycystic ovary syndrome. MATERIAL AND METHODS: The study was performed at the Ward of Obstetrics and Gynecology of the General Hospital in Subotica. A retrospective investigation comprised 39 patients with polycystic ovary syndrome. All patients were in the fertile age-range: 18-38 years. Following ultrasonographic examination and anamnestic data, patients underwent hormonal analyses of follicle-stimulating hormone (FSH), luteinizing hormone (LH), testosterone, prolactin and insulin obtained from the pooled serum sample. RESULTS: Values of testosterone and insulin in the group of obese patients with polycystic ovary syndrome were significantly higher than in normal weight patients. Patients with normal body weight index had significantly increased levels of LH in regard to those with increased body weight index. Values of FSH, prolactin and LH/FSH ratio were not significantly different in both groups of polycystic ovary syndrome patients. Increased values of insulin were recorded in 43% of obese and 18.2% of normal weight patients. CONCLUSION: Analysis of investigated results confirmed that obese patients with polycystic ovary syndrome and insulin resistance have been a special clinical entity, whereas an open question remains whether obesity is directly connected with polycystic ovary syndrome or it is only an additional factor interfering with metabolic and hormonal status of genetically predisposed and phenotypically indoctrinated women with polycystic ovary syndrome. PMID- 14740541 TI - [Total thyroidectomy in C-cell hyperplasia of the thyroid gland]. AB - INTRODUCTION: Medullary thyroid carcinoma (MTC) is a neuroendocrine tumor of the thyroid parafollicular or C-cells. MTC accounts for approximately 3 to 5% of thyroid carcinomas. A characteristic feature of this tumor is production of calcitonin. Sporadic MTC accounts for 60-80% of all cases of the disease. The most common presentation of sporadic MTC is a solitary thyroid nodule, which occurs in 75 to 95% of patients. It typically occurs in the fifth or sixth decades, with slight female predominance. CASE REPORT: A case of a 58-year old woman with elevated basal calcitonin levels (221 pg/ml) has been presented. Preoperative ultrasound of the thyroid gland and body (99m)Tc DMSA scintigraphy showed a diffuse goiter. Hyperparathyroidism and pheochromocytoma were not present. Total thyroidectomy was performed. Only C-cell hyperplasia was found. DISCUSSION AND CONCLUSION: Many patients with elevated basal serum calcitonin level ranging between 150 and 1000 pg/ml, have only C-cell hyperplasia, but a few have small MTCs. For persons in known kindreds with familial MTC, prospective screening, using serum calcitonin measurement, can identify those at risk for the disease before MTC is clinically evident. Due to the possibility that any patient with MTC may have multiple endocrine neoplasia type 2 (MEN-2), preoperative testing must also include measurement of serum calcium (to rule out hyperparathyroidism) and a test for pheochromocytoma. Total thyroidectomy is indicated in patients with high levels of calcitonin. PMID- 14740542 TI - [Punishment--"the work of God"--forensic accounts of crime and punishment]. AB - INTRODUCTION: This paper deals with coincidental deaths of persons previously engaged in violent behaviour. In victimology, violent deaths rest on legal authorities and public prosecutor. In some cases the clearance of circumstances is "sui generis", as a consequence of "vis maior". However, in some cases, depending of circumstances, perpetrators are uncovered without further investigations. CASE REPORT: This paper presents some of the most interesting cases from our Forensic practice: infanticides, "jealousy" killing, other murders. All crimes were recovered and sanctioned. History is full of cases like these described. Man is the only primate who kills and tortures other men. Nothing can fill the soul as hatred. Hatred is stronger than love. Love can easily lead to hate. Man is afraid of being killed, but often acts like a victim. The man proves himself worthy through his life, not death. PMID- 14740543 TI - [Confocal microscopy in transplanted human corneas]. AB - PURPOSE: The aim of the study was in-real time observation and morphological evaluation of the transplanted human corneas, using confocal 2.0 microscopy. METHODS: Twelve eyes of 12 patients on 3-7 days after lamellar keratoplasty (LKP), 8 eyes of 8 patients at 1 year after LKP, and 10 eyes of 10 patients at 3 7 days after penetrating keratoplasty (PKP), 11 eyes of 11 patients at 1 year after PKP were examined by confocal 2.0 microscopy in vivo. Images were recorded by continuously focusing the optical section through the full thickness central cornea. RESULTS: 3-7 days after LKP, small stromal cells, cranny-like dark strias and nerves were seen in transplanted corneas. There were highly reflective regions and dots in the lamellar interface. One year after LKP, the nerves disappeared. There were less highly reflective regions, but the dots still can be seen. Some gross dark strias were seen in the posterior stroma and the density of the endothelium cells was normal. 3-7 days after PKP, keratocytes were activated. Nerves and gross dark strias could be seen in transplanted corneas. The density of the endothelium cells was normal, with some highly reflective dots deposited among them. One year after PKP, the nerves disappeared, and strias still existed. The density of the endothelium cells decreased significantly. CONCLUSIONS: Confocal scanning microscopy is a new tool for the study of cellular morphology and construction of transplanted human corneas. It can be used in the evaluation of operation effect, clinical observation and follow-up. PMID- 14740544 TI - [Study of diopters-cutting in treatment of PRK for myopia]. AB - PURPOSE: To study a method of determining the diopters-cutting for photorefractive keratectomy (PRK) pre-operation. METHODS: By statistics, to analyze the differences of diopter between optometry of mydriasis and keratectomy in 143 cases 209 eyes of medium myopia with complete data recorded. RESULT: The average diopter for optometry of mydriasis is -4.32 D, for keratectomy, -4.74 D. The average of differences is (0.42 +/- 0.173) D, P < 0.01. CONCLUSION: The numerical value of diopter-cutting for operation should be higher than for optometry of mydriasis in treatment of PRK for myopia. PMID- 14740545 TI - [Refractive error and the intraocular pressure: findings in the Chinese eyes]. AB - PURPOSE: To analysis the correlation between the refractive error and the intraocular pressure. METHODS: The IOP was measured in 142 eyes with noncontact tonometer. The refractive error were determined with an autorefractometer(TOPCON 7100). RESULTS: The analysis of variance showed that IOP of medium myopia is the highest, and a significant IOP difference between medium myopia and severe myopia, and IOP of patients less than 19yrs is significantly higher than patients more than 30 yrs; and there is also no significant influence of gender on patients' IOP, mean IOP(13.04 +/- 2.97) mmHg. CONCLUSION: These results suggest that when medium myopia IOP is relatively higher, or when a young patient less than 19 whose IOP is relatively higher than usual, that may mean the eye is under an unstable situation. PMID- 14740546 TI - [Affection of diopters and fundus pathologic changes in high myopic eyes]. AB - PURPOSE: To investigate the relations of the faulty vision correction caused by high myopia with the increase of diopters and progress of fundus retrograde affection. METHODS: One hundred and forty cases (280 eyes, diopters are -6.25 30.00) were examined with retinoscopy after undergoing mydriasis. Then the axial length of the anterioposterior diameters was measured with A-wave and B-wave ultrasonography. The fluorescein fundus angiography (FFA) was performed on these eyes. The data were statistically analyzed. RESULTS: The faulty vision correction of high myopic eyes was obviously decreased with the increase of the diopters. When the visual acuity was -20.00 D, the average correction was clinically lower than 0.42 D. And it could only reach 0.25 D, with the vision being -25.00 D. The difference is very significant (P < 0.001). FFA showed that the pathologic changes of fundus posterior pole appeared with the different degrees of damages, such as simple lesion, diffuse lesion, psoriasis lesion and complex lesion. And the vision correction could be simultaneously influenced by multiple complications. The difference is of significance (P < 0.01). CONCLUSION: Vision correction in high myopic eyes is obviously decreased with the increase of diopters. The higher the diopter is, the harder the vision can be ideally corrected. The greater the pathologic change at the fundus posterior pole, the severer the degree of the damage. The more obvious the visual function is influenced, the harder the vision can be corrected. This paper revealed that high myopia is a kind of complex eye disease. It can be treated not only by lens correction but also by other combined treatments. PMID- 14740547 TI - [Vitreous hemorrhage and vitreoschisis]. AB - PURPOSE: To identify the morphologic characteristics of vitreous especially vitreoschisis in vitreous hemorrhage, aiming at discussing the mechanism and clinical importance of vitreoschisis. METHODS: Eighty-eight cases (97 eyes) of vitreous hemorrhage were observed among which 32 cases (38 eyes) resulting from PDR, 28 cases (30 eyes) resulting from retinal vasculitis and 28 cases (29 eyes) resulting from retinal vein occlusion. All cases had B-scan examination, some had color Doppler examination. Images were compared with the finding in vitre-ctomy. RESULTS: Vitreoschisis occurred in 18 cases (47.4%) in PDR group, 10 cases (33%) in retinal vasculitis group and 4 cases (13.8%) in retinal vein occlusion group. CONCLUSIONS: Vitreoschisis may occur in different kind of vitreous hemorrhage. Recognition of vitreoschisis may aid to complete vitrectomy and reduce some complication after surgery. PMID- 14740548 TI - [Scoring method evaluation for threshold visual field defect in glaucoma]. AB - PURPOSE: To evaluate the validity and reliability of visual field defect scoring method in glaucoma. METHODS: The visual field defects of 91 glaucoma cases (91 eyes) were scored with the Advanced Glaucoma Intervention Study (AGIS) modified scoring method, and analyzed the relation and correlation between the score and cup/disk ratio, mean defect (MD) and loss variance (LV). RESULTS: The larger the score was, the larger the cup/disk ratio, MD and LV were. The significant positive correlation was found between the score and the cup/disk ratio (r = 0.8712), and the correlation coefficient was larger than that of between the MD, LV and cup/disk ratio. CONCLUSION: The visual field defect score could more accurately reflect the situation of glaucomatous optic nerve damage than visual field indices, and could quantify the visual field damage extent in glaucoma. PMID- 14740550 TI - [Analysis of the methods and effects of cataract surgeries in the poor areas]. AB - PURPOSE: To evaluate the effective method for the vision rehabilitation for cataract patients in the poor countryside in mountain areas. METHODS: To review and analyze the method by the Lee Karsing Foundation for vision rehabilitation since November 1998. RESULTS: Until December 2002, 10,569 people got free treatment, 1,164 cataract patients regained their vision after free surgery. CONCLUSIONS: That we serve the patients with safe and effective surgery and call on the spirit of devotion as well as safe medical resources, is one of the aims of our job in the poorly equipped hospitals for the "Vision Recovery & Poverty Aid Event". PMID- 14740549 TI - [Application of endolaser in vitrectomy for retinal vascular disease]. AB - PURPOSE: To discuss the usage and effectiveness of endolaser in vitrectomy for retinal vascular disease. METHODS: Twenty-four cases (28 eyes) of vitrectomy for retinal vascular disease were treated with endolaser. The methods of endolaser: 21 eyes were treated with panretinal scatter therapy; 2 eyes with endolaser ring intercepting around retinal and sealing retinal breaks; 5 eye with focal therapy. RESULTS: After follow-up for 1-24 months (mean 9 months), retinal hemorrhage recurred in 3 eyes; the other 25 eyes, retinas were totally reattached, retinal breaks were sealed, and vision had considerable recovery, no hemorrhage. The ratio of recovery is up to 89.2%. CONCLUSIONS: Endolaser is an important adjuvant in vitreous operation nowadays and an effective procedure in vitrectomy for the cases of retinal hemorrhage. PMID- 14740551 TI - [Application of endoscopic operation in ophthalmology]. AB - The application of endoscopic operation in ophthalmology has gotten great progression in recent years. It has showed many advantages such as more effectiveness, reliability and less damage, complications. Ophthalmic laser endoscopic operations can be applied in the management of glaucoma, vitreoretinopathy and so on. Lacrimal endoscopy in the treatment of lacrimal apparatus disease and nasal endoscopy in the Ophthalmo-Rhinol associated surgeries also has got great development. The application of ophthalmic endoscopic operation has indicated that the period of mini-wounded surgery in ophthalmology began. In this paper, we will describe the basic principle, status of application and prospect of endoscopic operation in ophthalmology. PMID- 14740552 TI - [Study on effects of interleukin-1 and their inhibitors for proliferation and hyaluronic acid synthesis of cultured human orbital fibroblasts]. AB - PURPOSE: To observe the proliferation and synthesis of hyaluronic acid (HA) of cultured human orbital fibroblasts (OFs) affected by interleukin-1 (IL-1), and their inhibitors interleukin-1 receptor antagonist(IL-1Ra). METHODS: OFs from 5 patients with thyroid-associated ophthalmopathy (TAO) and 4 normal subjects were cultured in vitro. IL-1, IL-1Ra with different concentrations were put into media respectively, and IL-1(500 U/ml), IL-1Ra(5, 50 ng/ml) were put into media together. The proliferation and HA production of OFs were measured by MTT and radioimmunological technique. RESULTS: IL-1(550,500 U/ml) can significantly increase the proliferation of OFs and synthesis of HA (P < 0.05), IL-1 (500 U/ml) stimulate more proliferation of OFs and HA production of OFs from TAO patients than that of OFs from normal subjects. While IL-1Ra alone had no inhibitive effects at any concentrations. IL1Ra (5.50 ng/ml) can block the proliferation and HA production of OFs when IL-1 with IL-1Ra incubated together. CONCLUSION: IL-1 can stimulate the proliferation and HA synthesis of OFs. IL-1Ra is an effective inhibitor of proliferation and synthesis of OFs induced by IL-1. IL-1Ra may be useful in the prevention or treatment of TAO. PMID- 14740553 TI - [Role of p38MAPKs pathway in the growth inhibition of rabbit lens epithelial cells induced by EGCG]. AB - PURPOSE: Study the role of p38MAPKs pathway in the growth inhibition of rabbit lens epithelial cells(LECs) induced by green tea extract (-)-Epigallocatechin-3 gallate (EGCG). METHODS: (1) MTT colormetric assay was used to study the effect of p38 specific inhibitors SB203580 on LECs growth. (2) MTT colormetric was used to study the effect of SB203580 on LECs growth inhibition induced by EGCG. (3) Western blotting methods was used to study the effect of EGCG on the kinase phosphorylation- and nonphosphorylation-level of p38. RESULTS: (1) When LECs were preincubated with 25 mumol/L, 50 mumol/L SB203580 for 1 h, 100 mumol/L EGCG had little influence on LECs proliferation. It showed great protects effect significantly when increased to 200 mumol/L. (2) Basic phosphorylation-level of p38 were very weak in LECs, but it increased with EGCG concentration increased and reached maximum at 200 mumol/L. 15 min after 200 mumol/L was added, the phosphorylation-level was the highest and began to fall, but even at the end of our test, it still stayed higher than that of basic level. The nonphosphorylation level of p38 remained stable in all test. CONCLUSIONS: The LECs proliferation inhibition induced by EGCG is through p38 pathway at lease partially. PMID- 14740554 TI - [Change of alpha-crystallin acting as molecular chaperone activity with ageing]. AB - PURPOSE: To evaluate the molecular chaperone function of alpha-crystallin with ageing. METHODS: alpha-Crystallin of newborn, adult and old rabbits lenses in both of cortex and nucleus were separated by chromatography on Sephacryl S-300HR. The protection of alpha-crystallin against thermal aggregation of catalase and beta L-crystallin (60 degrees C), inactivation of catalase by fructose(37 degrees C) and heat stress(60 degrees C) were measured spectrophotometrically. RESULTS: Protection of alpha-crystallin against aggregation and inactivation using four methods showed a similar pattern. The protective ability in cortex was greatly higher than in nucleus of different-aged lenses, and alpha H-crystallin was less than alpha L-crystallin in both cortex and nucleus. There was no statistically decrease with age of chaperone function of both alpha H-crystallin and alpha L crystallin in the cortex, whereas alpha L-crystallin in the nucleus was compromised. CONCLUSION: alpha-Crystallin in the nucleus shows age-related decrease in chaperone function, which may be responsible for cataract formation. PMID- 14740555 TI - [Expression of TGF-beta receptor II in periretinal membranes of proliferative vitreoretinopathy]. AB - PURPOSE: To investigate the expression of transforming growth factor-beta receptor II (TGF-beta R II) in periretinal membranes of proliferative vitreoretinopathy (PVR). METHODS: Immunohistochemistry and in situ hybridization were used to detect the expression of TGF-beta R II protein and mRNA in sixteen proliferative membranes of thirteen PVR patients. RESULTS: Immunohistochemically staining showed that membranes labeled negative "-", weak positive "+", positive "2+" and strong positive staining "3+" were respectively noted one, one, three and four membranes in nine C2-C3 membranes. Total positive cell rate is 88.9%. Among seven D1-D3 membranes, one membrane was labeled "-", three were "+" and another three membranes were labeled "2+". Total positive cell rate is 85.7%. Positive staining cells were of a type of epithelial-like cells which frequently found pigment in or around them. Relation between TGF-beta R II expression and membrane grades appeared no correlation (P > 0.05). Results of in situ hybridization were almost consistent with that of immunohistochemistry. CONCLUSIONS: Both TGF-beta R II protein and mRNA are highly expressed and up regulated in PVR membranes by cytokine stimulation, moreover TGF-beta may play important role in the pathogenesis of PVR. PMID- 14740556 TI - [Correlation between secretion of VEGF and proliferation of cultured bovine retinal pigment epithelium cells under hypoxic condition]. AB - PURPOSE: To investigate the effect of hypoxia on bovine retinal pigment epithelial (RPE) cell secreting vascular endothelial growth factor (VEGF) and the correlation between VEGF and proliferation of the cultured bovine retinal pigment epithelial cell. METHODS: Cells we harvested were divided into different groups with or without anti-VEGF monoclonal antibody (VEGF MAb), and then grew under hypoxic and normal (as a control) condition. After 24 h, cell supernatants were collected and used for ELISA VEGF protein assay. The proliferation of cells grown under different conditions was detected by MTT colorimetric assay. RESULTS: Cell proliferation and concentration of VEGF protein: The hypoxia with VEGF MAb group (Group A) was (1.812 +/- 0.068), (55.07 +/- 19.18) pg/ml; the hypoxia without VEGF MAb group (Group B) was (2.292 +/- 0.197), (171.61 +/- 16.02) pg/ml; the normal with VEGF MAb group (Group C) was (1.350 +/- 0.185), (43.92 +/- 21.39) pg/ml; the normal without VEGF MAb group (Group D) was (1.435 +/- 0.157), (48.51 +/- 24.73) pg/ml. Group A was lower than Group B on cell proliferation and concentration of VEGF protein. There is obvious difference between the two groups on both sides (P < 0.05). Group A was higher than Group D on cell proliferation. There is obvious difference between them on cell proliferation (P < 0.05). Group B is higher than Group D on cell proliferation and concentration of VEGF protein. There is obvious difference between them on both sides (P < 0.05). There is a linear association between the concentration of VEGF protein and the proliferation of RPE cell under hypoxic condition by Pearson Correlation (r = 1.000, P < 0.05). CONCLUSION: Hypoxia stimulates the secretion of vascular endothelium growth factor of bovine RPE cell, which contributes to the proliferation of bovine RPE cell. PMID- 14740557 TI - [Multifocal electroretinogram in cystoid macular edema]. AB - PURPOSE: To evaluate the visual function in different area at retinal posterior pole for the patients with cystoid macular edema (CME). METHODS: Multifocal ERG was tested using visual evoked response imaging system (VERIS) 4.0 for 24 patients (27 eyes) with CME and 25 normal control subjects whose ages matched patient's ages, the traces and 3D function maps of first order kernel response were recorded on 103 retinal locations of 26.6 degrees X22.1 degrees visual field. RESULTS: The latencies of N1 and P1 waves were remarkably prolonged at whole area and the 6 rings in the CME, there were statistically significant differences between the normal control group and CME (P < 0.01). Compared with normal control group, the average response densities of N1 wave at 1, 2, and 3 rings in the CME were seriously reduced (P < 0.01), but they in 4, 5 and 6 rings were not significantly changed (P > 0.05), the average response densities of P1 at whole area and the 6 rings were reduced (P < 0.05). CONCLUSIONS: The mfERG is a promising tool to evaluate the visual function in different area at retinal posterior pole for the patients with CME. The response densities of N1 wave are better analysis index for CME. PMID- 14740558 TI - [Comparison of multifocal electroretinogram in six kinds of maculopathies]. AB - PURPOSE: To compare the differences of multifocal electroretinogram (mERG) in six kinds of macular diseases. METHODS: The mERGS of 6 kinds of macular diseases were recorded and compared with normal corresponding age groups. RESULTS: The differences of mERG between maculopathies and control were significant. The amplitude densities of N1 and P1 of all 6 rings were remarkably decreased, the latencies were also significantly prolonged in all kinds of macular diseases(P < 0.01). The amplitudes in central 1 ring were lowered most seriously comparing with other rings. There was a tendency that the differences of amplitudes of mERG between the maculopathies and normal groups were decreased as the acentric degree increased. In the 6 kinds of macular diseases, the 1 ring amplitudes of mERGs of exudative age-related macular degeneration (ARMD), central exudative chorioretinitis, idiopathic macular hole and Stargardt's disease were more seriously decreased than those of atrophic ARMD, idiopathic epiretinal macular membrane and idiopathic central serious chorioretinpathy. CONCLUSIONS: mERG is an effective method to test macular function. There are group differences of macular function between different kinds of macular diseases. PMID- 14740559 TI - [Characteristics of normal infants' flash electroretinograms]. AB - PURPOSE: To investigate the characteristic of retinal function of Chinese infants with ages ranging from 4 to 5 months. METHODS: The flash electroetinograms(ERGs) of 17 infants(34 eyes) of 4 months, 13 infants (26 eyes) of 5 months and 39 adults (78 eyes) were tested. Their rod responses, cone responses and maximal combined responses were recorded. The amplitudes and latencies of b waves were compared and analyzed among the 2 infant groups and the adult group. RESULTS: In each of 2 infant groups, of all 3 responses, the means of amplitudes were significantly lower and latencies were significantly longer than those of adults. Between the 2 infants groups, rod responses and maximal combined responses were not significantly different, but the cone responses were significantly different, the infants group with age of 5 months has higher amplitude and shorter latency. The average amplitudes of the rod responses of the 4 months infant group and the 5 months infant group were 38.9% and 36.7% of that of the adult group respectively; the average amplitudes of the cone responses of the 4 months infant group and thd 5 months infant group were 62.6% and 71.5% of that of the adult group respectively. CONCLUSIONS: Between the ages ranging from 4 to 5 months, all of the responses of the infant ERG had not reached the levels of adult. Cone response developed more rapidly and was more mature than rod response. PMID- 14740561 TI - [Pesticide and central nervous system]. PMID- 14740560 TI - [Care and nursing cooperation during laser cataract surgery]. AB - PURPOSE: To discuss the care and nursing cooperation during laser cataract surgery. METHODS: The care and nursing cooperation of 70 cases (70 eyes) of laser cataract surgery were retrospected. Forty-one cases were male and 29 cases were female. The age ranged from 36 years to 75 years. RESULTS: All operations were completed smoothly with the nursing cooperation. The patients showed good cooperation during surgery with the nursing care. Visual acuity had been improved significantly after the surgery. CONCLUSIONS: To assure the surgery performed smoothly, nurses of the operating room must know about the proper operation of the laser phacolysis machine and the procedure of the surgery. Careful preoperative preparation and meticulous intra-operative care are also very important. PMID- 14740562 TI - [Advanced hazmat life support (AHLS). Outline of the system]. PMID- 14740563 TI - [Advanced hazmat life support(AHLS). Provider course]. PMID- 14740564 TI - [A proposal guideline for Japanese version of AHLS]. PMID- 14740565 TI - [Active involvement of pharmacists in initial treatments for acute poisoning and overdosed patients in the intensive care unit]. AB - We have developed guidelines of pharmaceutical activity which describes how pharmacists are actively involved in supporting the initial treatment of poisoning and overdosed patients in the Intensive Care Unit (ICU). These guidelines are derived from the original procedural manual that consisted of protocol charts. The charts provide the pharmacist including ICU staff members with directions for the collection of clinical information and the forms to use for documentation. We focused on appropriate collection and proper preservation of collected samples in order to perform diagnostic analysis when needed. The original Information Record Form, completed by the participating pharmacist, documents all information regarding the patient's care. This record provides for integration of the diverse and complicated clinical information of the patient so that the physician can gain a comprehensive picture of the patient. The guidelines, manual book, materials for sample collection and the Information Record Form are stored in a container called the "Poisoning Aid Set", which will be available in the ICU. The guidelines are activated once an emergency call from paramedics is received regarding a suspected poisoning patient. According to the guidelines, the emergency room (ER) physician immediately contacts the pharmacist who will provide his professional services as a member of the treatment team. We have applied these guidelines to 29 poisoning patients and have critically evaluated for effectiveness. Early participation by pharmacists, by reviewing timely, accurate and competent clinical information enabled the pharmacist to identify the suspected drug from the biological samples obtained. From our experiences, we conclude that this active involvement of pharmacist in the initial treatment of poisoning and overdosed patients in the ICU was both supportive and beneficial to the patient. In addition, the participation of pharmacist as a member of a treatment team provided an excellent opportunity to collaborate with the entire ICU staff members. PMID- 14740566 TI - [Determination of amanitatoxins by HPLC]. AB - High-performance liquid chromatographic (HPLC) assay has been developed for the simultaneous determination of alpha-amanitin, beta-amanitin and phalloidin in serum. Three toxins were extracted by reflux in a water bath at 80 degrees C for one hour and purified by Sep-Pak Plus tC18 cartridges. The HPLC assay was performed under gradient conditions using Develosil RP AQUEOUS column. The moble phase consisted with a mixture of acetonitorile containing 0.01 M ammonium acetate(pH 5.0). The column effluence was monitored at 295 nm, 302 nm and 230 nm for 35 min. Detection limit of three toxins in serum were 0.2 microgram/ml respectively. High recovery yields in the range of 81.5-88.1% for toxins were obtained by using this method. PMID- 14740567 TI - [Intestinal edema caused by ingested formalin]. AB - A 65-yr-old man treated for depression was transferred to our hospital. He ingested over 150 ml of formalin for suicidal attempt. On admission, he was hypotensive, developing acute renal failure and liver dysfunction. During first 36 hours, he needed 21 l of lactate Ringer solutions to maintain enough urination. The abdominal computed tomography showed obvious edema of intestine and endoscopic findings of the upper digestive tract were corrosive erosions. We started proton pump inhibiter on the first day. After a week, they changed in many ulcers with smooth-surfaced elevation on the greater curvature of the stomach. On the 15th hospitalized day, he was discharged from our hospital without sequelae. Formalin consists of forty percent solution of formaldehyde and methanol. Ingestion of formaldehyde may cause burning in the digestive systems and harmful effect to major organ such as kidney or liver. Although previous reports said that formalin has direct toxicity to major organs, we could discharge him without sequelae by treatment with large amount of fluid resuscitation. PMID- 14740568 TI - [Case report-fatal snail bait (metaldehyde) overdose presenting aspiration pneumonia]. AB - A-55-year-old man ingested unknown amount of snail poison bait containing metaldehyde. He was mentally retarded and presented pica. On admission, his vital sign was stable, and the extremeties were spastic. Then, gastric lavage was unsuccessful because of massive unbited food. Activated charcoal and cathartic were administrated. On the next day, general convulsion occurred and respiratory distress advanced, so he was intubated. On the 3rd day, infiltration shadow appeared on chest roentogenogram and, his respiration was assisted mechanically. Thereby, acute lung injury advanced regardless of tracheostomy, kinetic therapy, antibiotics and steroid pulse therapy. He died of respiratory failure on the 33rd day. Serum test showed HBs and HBe antigen, CT scan revealed ascites and splenomegaly; the clinical course might be worsened by liver cirrhosis. HPLC revealed metaldehyde in the serum (total 80.6 microg/ml). He ingested 2.7 g of metaldehyde maximally estimated. Although Japan Poison Information Center reported that snail poison bait poisoning is often in dogs in Japan, human poisoning is rare. PMID- 14740569 TI - [A case of mixed poisoning by organophosphate and methanol]. AB - We encountered a case of mixed poisoning by organophosphate and methanol. Each poisoning was comparatively frequent but we reviewed no cases of mixed poisoning of them. A 49-year-old woman was transferred to our hospital because of oral ingestion of organophosphorous compound (about 14 g of fenitrothion) and glass cleaner (about 40 g of methanol) for suicidal purpose. She underwent general antioverdose treatment including gastric lavage, activated charcoal and cathartics. For fenitrothion poisoning, she received atropine and pralidoxime. For methanol poisoning, she was treated with hemodialysis. Three days later, she moved to psychiatric ward from emergency ward without aftereffects and was given a diagnosis of depression. PMID- 14740570 TI - [Two cases of "Ecstasy (MDMA)" poisoning]. PMID- 14740571 TI - [Practical analysis of toxic substances useful for clinical toxicology-8 paraquat]. PMID- 14740572 TI - [Commentary on the standard therapeutic practice of poisoning recommended by Japanese Society for Clinical Toxicology--3--whole bowel irrigation]. PMID- 14740573 TI - [2002 annual-report of mass casualty incidents]. PMID- 14740574 TI - Nursing management of patient with Pericardial effusion. PMID- 14740575 TI - Nurse: the key co-ordinator for patient care in hospital. PMID- 14740576 TI - A study to understand and evaluate the nursing care of low birth weight babies. PMID- 14740577 TI - Leading and succeeding in outcomes management. AB - An effective OM program offers the best in clinical care, efficiency, and use of organizational resources and results in overall satisfaction for health care consumers, payers, and providers. Although challenges abound in today's health care environment, a strong interdisciplinary team with the appropriate resources and supports and led by a skilled outcomes manager will succeed. Such a team will also develop as other needs and opportunities exist in the future. PMID- 14740578 TI - Symptom management intervention in elderly coronary artery bypass graft patients. AB - The purpose of this pilot study was to test the impact of a symptom management intervention (using a device called the Health Buddy) on recovery outcomes (symptom evaluation and response and postoperative problems) of elderly coronary artery bypass graft patients at time of discharge, at 2, 4, and 6 weeks after surgery, and at 6 months after surgery. While there were no statistically significant differences found for many of the study variables, there were trends in the intervention group of more improvement for many of the outcome variables, supporting the need for a randomized clinical trial. PMID- 14740579 TI - Nurses' integration of outcomes assessment data into practice. AB - The purpose of this article is to provide initial insight from rational and phenomenologic theoretical perspectives into how nurses integrate baseline and follow-up outcomes assessment data into practice to inform their clinical decision-making. Preliminary findings from 29 nurse interviews indicate that some nurses use outcomes assessment data for evaluating their interventions and for decisions concerning initial and/or discharge planning. Nurses' perceived usefulness of outcomes data was based on task complexity and nurses' level of expertise. PMID- 14740580 TI - Clinical and fiscal outcomes of utilization review. AB - Concurrent utilization review (UR) is both a quality improvement tool and a cost containment strategy used by managed care organizations. The UR process requires that providers (hospital staff) communicate clinical information about hospitalized patients to payers who evaluate the appropriateness and medical necessity of the planned care. Payers then make a decision whether to certify the care for reimbursement. This study provides data to indicate that denials of certification have little impact on clinical and fiscal outcomes of patient care. PMID- 14740581 TI - Reinforcing organizationwide pressure ulcer reduction on high-risk geriatric inpatient units. AB - Skin care and pressure ulcer prevention programs abound, although their content varies and their outcomes are often difficult to quantify. This article describes 2 complementary programs, their quality improvement processes, and a variety of ways of measuring their success. The first program was broad in scope, emphasizing system-wide changes in administration and coordination of resources, while the second focused on nursing education on high-risk units. These 2 approaches could be adapted for use in any health care setting. PMID- 14740582 TI - National databases and clinical practice specialist: decreasing postoperative atrial fibrillation following cardiac surgery. AB - In this article, we document how an interdisciplinary committee of health professionals led to an approximate 50% reduction in the incidence of postoperative atrial fibrillation (AF) following a cardiac surgery procedure by using preoperative loading and dosing of PO amiodarone and beta blockade. Patients in this report (n = 3397) included all coronary artery bypass surgery (CABG) and valve replacement/repair procedures from January 1, 2000 to June 30, 2002. The incidence of postoperative AF for a CABG or valve replacement/repair procedure was 19.0% for period A (preprotocol) and 13.5% for period B (postprotocol). This translates into an absolute risk reduction of 5.5% or 113 actual cases of postoperative AF reduced. The role of the clinical practice specialist, a master's prepared nurse, and participation in the national Society of Thoracic Surgeon's database allowed us to track our CABG outcomes, benchmark our outcomes against both national and regional institutions, and make changes in outcomes incidence through performance improvement. PMID- 14740583 TI - Heart failure management across the continuum: a communication link. AB - Patients with heart failure often become lost in the ambulatory healthcare maze on hospital discharge. This causes fragmentation of care due to a lack of a communication link with the ambulatory setting. This article discusses a quality improvement project and addresses the use of a communication tool that helps to transition the plan of care for the heart failure patient from acute care to the ambulatory care setting. The key points focus on the continuance of the plan of care that began during hospitalization and is then extended into the ambulatory setting through care management services. PMID- 14740584 TI - Postdischarge outcome rates influenced by comorbidity and interdisciplinary collaboration. AB - This study investigated the relationship between health care professionals' ratings of their discharge planning collaboration and other patient characteristics with rates of postdischarge outcomes experienced by a sample of elderly veterans. The significant influence of interdisciplinary collaboration levels and comorbidity was demonstrated on emergency room visits and postdischarge fall rates. The author discusses several complex methodological issues for future studies on the relationship between programmatic and patient characteristics and cross-sector patient outcomes. PMID- 14740585 TI - Hospitalized elders: changes in functional and mental status. AB - This article describes outcomes of a new model of care for hospitalized elders and their families. Patient functional and cognitive status on admission and discharge were evaluated for changes as a result of an educational program for preparing family-centered geriatric resource nurses. Patients in the intervention group (n = 173) demonstrated significant improvements in outcome measures (functional and cognitive status) from admission to discharge. A subset (n = 50) was selected from the 173 subjects who comprised the intervention group; this subset was compared with control subjects (n = 44); no statistically significant differences were noted between the 2 groups. Suggestions for future research are presented. PMID- 14740586 TI - Academic nurse-managed centers: approaches to evaluation. AB - A comprehensive evaluation plan was developed to assess the outcomes of a multiuniversity project to support the development of academic nurse-managed centers (ANMCs). The evaluation included measuring ANMC impact on the clients and communities served, on students, and on the sponsoring faculties and universities. This article includes a discussion of the processes used in evaluation plan development, the variables measured, the tools developed to measure selected variables, and a summary of evaluation findings. Recommendations for use of selected evaluation components across ANMCs are presented. PMID- 14740587 TI - The spongia somnifera. PMID- 14740588 TI - Perianesthetic management of patients undergoing resection of pheochromocytoma. PMID- 14740589 TI - Bilateral interpleural versus lumbar epidural bupivacaine-morphine analgesia for upper abdominal surgery. AB - This randomized study was designed to compare the effectiveness of bilateral interpleural analgesia with lumbar epidural analgesia, on postoperative pain relief in upper abdominal surgery. The studied patients were randomely allocated into either interpleural group "IP" (n = 15) or epidural group "EP" (n = 15). In "IP" group, preanesthetic bilateral interpleural block was done using a mixture of bupivacaine 0.5% (0.8 mg/kg) and 2 mg morphine diluted to 50 ml saline for each side. In "EP" group, the same mixture-diluted in 20 ml saline-was injected in the epidural space (L2-3). The general anesthetic technique was the same in both groups. Hemodynamic, gasometric, verbal pain score (VPS) values and complications were compared in both techniques. Heart rate (HR) and mean arterial pressure (MAP) readings were in the accepted normal range in the perioperative period although significant lower readings were detected in "EP" group. No significant differences were displayed in blood gasometric variables between the two groups. There were considerable level of analgesia in both groups in the postoperative period although "EP" analgesia was superior to "IP". More pain free patients (9 versus 4) and significant lower consumption of nalbuphine were detected in "EP" group. The results of this study indicate that bilateral "IP" analgesia may offer a satisfactory analgesia for upper abdominal surgery when the use of other analgesic techniques may be contraindicated. PMID- 14740591 TI - Effects of different disinfectants on decontamination of laryngoscopes. AB - Guidelines for controlling possible contamination of laryngoscopes should be formulated with the benefit of relevant experimental data. In this study, the effects of five different disinfectants commonly used for the disinfection of laryngoscopes are evaluated. We formed 14 groups, with 10 blades in each. The first 7 groups were contaminated with hospital related meticillin resistant Staphylococcus aureus (MRSA), and the remaining 7 groups with hospital related multiple resistant Pseudomonas aeruginosa (PA). For the first group of blades, no disinfection procedure was carried out and, were assumed as a control group. Blades in remaining groups were rested for 10 minutes in containers containing 70% alcohol (II), 1/100 dilution of cetrimide (III), 1/100 dilution of chlorhexidine (IV), 1/10 dilution of chlorhexidine (V), 1/10 dilution of povidone iodine (VI), and 1/100 dilution of ammonium chloride (VII). Disinfectant used in a group was considered effective when growth was seen in 5 or less than 5 plates representing that group. All disenfectants tested were found effective on decontamination of laryngoscopes. Five different moderate level disinfectants, which are commonly used for the disinfection of laryngoscopes, have been found effective even on resistant hospital microorganisms like MRSA and P. aeruginosa. They may be the choices of the disinfectants, especially 1/10 dilution of chlorhexidine gluconate and 1/100 dilution of ammonium chloride. PMID- 14740590 TI - Effects of sevoflurane versus TIVA on gastric intramucosal pH and hemodynamic status in colon cancer surgery. AB - BACKGROUND: Gastric tonometry is a minimally invasive device by which the adequacy of splanchnic blood flow is assessed indirectly. Patients undergoing major abdominal surgery are under the risk of developing splanchnic ischemia. The aim of this study was to compare gastric intramucosal pH and hemodynamic effects between sevoflurane and TIVA achieved by propofol. METHODS: Forty patients of ASA I-II were studied. Twenty patients received sevoflurane (Group S) and 20 patients received TIVA by propofol (Group P). Continuous measurements were made at four intervals: (a) 30 minutes before the starting of the surgery, (b) at the first hour of the surgery, (c) at the second hour of the surgery, (d) at the first hour after ending of the surgery. The mean arterial pressure, heart rate, SpO2, EtCO2, body temperature, central venous pressure, urine output, intramucosal and arterial pH were monitored. RESULTS: There were no significant differences between two groups regarding the studied parameters. CONCLUSIONS: Sevoflurane and propofol did not cause any significant difference in hemodynamic effects and splanchnic circulation measured by gastric tonometry in ASA I-II patients operated on for colon cancer. PMID- 14740592 TI - The protective effects of early treatment with propofol on endotoxin-induced acute lung injury in rats. AB - To investigate the effects of treatment with propofol administration at different time point in acute lung injury of endotoxin-induced shock rats. METHODS: 76 male wistar rats were randomly assigned to five groups: A) control group; B) endotoxemic group, receiving intravenous lipopolysaccharide (LPS) 8 mg.kg-1; C) pretreatment group, treated identically to endotoxemic group with the additional administration of propofol (5 mg.kg-1 bolus, followed by infusion at 10 mg.kg-1.h 1) of 1 hr prior to the injection of LPS; D) simultaneously treatment group, treated identically to endotoxemic group with the additional administration of propofol simultaneously with the injection of LPS; E) post-treatment group, which was treated identically to endotoxemic group except for administration of propofol 1 hr after the injection of LPS. PaO2, pH, MAP and survival rate were recorded and plasma NO, TNF-alpha were measured during 5-hr after the injection of LPS. After the rats were killed, lung tissue was sampled to measured expression of inducible nitric oxide synthase (iNOS), nitrotyrosine (NT), myeloperoxidase (MPO) activity, malondialdehyde (MDA), wet-to-dry lung weight ratio (W/D), and pulmonary permeability index (PPI). RESULTS: Compared with the endotoxemic group, both the pretreatment and simultaneously treatment groups, significantly improved PaO2, pH, MAP and 5th hour survival rate of rats, and attenuated endotoxin-induced increased iNOSmRNA, NT expression, MPO activity and MDA level in lung tissue, and decreased pulmonary microvascular permeability, TNF alpha, NO in plasma. But these beneficial efficacies were blunted in the post treatment group. CONCLUSIONS: These findings showed that propofol administration may provide protective effects on acute lung injury in endotoxin-induced shock. PMID- 14740593 TI - Effect of controlled hyperventilation on the pressor response to laryngoscopy and tracheal intubation. AB - Pressor response to laryngoscopy and tracheal intubation includes rises in blood pressure and heart rate. This response may be harmful in the presence of cerebral or myocardial diseases. Although different preventive measures have been developed the choice of the agent or method has not been defined clearly. Hypocapnia is commonly used in anesthesia practice for different indications. It depresses the cardiovascular system and lowers the cardiac output. This study investigated the effect of controlled hyperventilation on the pressor response to laryngoscopy and tracheal intubation in three groups of healthy adult patients with different levels of end tidal CO2. The blood pressure and heart rate were recorded during induction of general anesthesia before and after laryngoscopy and tracheal intubation. The pressor responses to laryngoscopy and tracheal intubation in hypocapnic and normocapnic groups were comparable. Moderate degrees of controlled hyperventilation caused relatively more fluctuation in blood pressure during induction of anesthesia. It can be concluded that controlled hyperventilation has no beneficial effect upon the pressor response to laryngoscopy and tracheal intubation. PMID- 14740595 TI - The relief of anxiety: comparative study--air plane flying vs anesthesia. AB - BACKGROUND: Airplane Flying and anesthesia are both not entirely safe. Passengers and patients have the right not to be endangered during flight or anesthesia. Flying and anesthesia has always been associated with anxiety in passengers and patients. OBJECTIVE: This study was undertaken to compare the anxiety between passengers to fly and patients to receive anesthesia. METHODS: One hundred and eighty persons were randomly selected and divided into six equal groups. Pilots and flight crew; anesthetist, resident of anesthesiology and nurse anesthetists; people who had flown but without any history of anesthesia; people without any experience of flight or anesthesia; people with previous experiences of both the flight and anesthesia and finally people with a previous history of anesthesia but without any experience of flight as Groups 1 to 6 respectively. A questionnaire was used to evaluate the level of anxiety during both the flight and anesthesia. RESULTS: All six groups had significantly more anxiety from anesthesia than flying (p < 0.05) except in group 5 (p = 0.460). Anxiety of flying was significantly less in pilots and flight crews (group 1) when compared with other groups (p < 0.004). The anxiety of anesthesia was significantly less in anesthetist team when compared with those who has not experienced general anesthesia. People who had not experienced anesthesia showed more fear about anesthesia than those who had experienced general anesthesia (p < 0.002). CONCLUSION: The results showed that having enough information about flying causes less anxiety just as preoperative visits help the patient to undergo a more comfortable anesthesia. PMID- 14740594 TI - Hypobaric bupivacaine spinal anesthesia for cystoscopic intervention: the impact of adding fentanyl. AB - BACKGROUND AND OBJECTIVE: Addition of fentanyl to hyperbaric bupivacaine spinal anesthesia prolonged the duration of sensory block. This study seeks to test the hypothesis that adding fentanyl to small dose hypobaric spinal anesthesia will improve intraoperative patients and surgeon satisfaction without delay in recovery. METHODS: Patients (n = 80) subjected to minor cystoscopic surgery were randomly assigned to have spinal anesthesia with either 5 mg bupivacaine 0.1% or 5 mg bupivacaine 0.1% mixed with 20 micrograms fentanyl. The main outcome measures included intraoperative patient and endoscopist satisfaction, sedative/analgesic supplementation, postoperative side effects and time to ambulation. RESULTS: Patients in the bupivacaine group needed more analgesic supplementation. Analgesia was more adequate in the bupivacaine-fentanyl group. Pruritus was the main side effect in the bupivacaine fentanyl group. Ambulation and discharge of patients were nearly the same in both groups. CONCLUSIONS: Spinal anesthesia with small dose (5 mg) hypobaric (0.1%) bupivacaine mixed with fentanyl (20 micrograms) produced adequate anesthesia for short cystoscopic procedures with minimal side effects and without delay in ambulation. PMID- 14740596 TI - Anesthetic management of a patient having a carcinoid syndrome. PMID- 14740597 TI - Continuous paraverte bral block for the management of post-thoracotomy pain in a patient undergoing tracheal reconstruction. PMID- 14740598 TI - Traumatic asphyxia: a case report and anesthetic implications. PMID- 14740599 TI - Kinking of epidural catheter--a case report. AB - Kinking of an epidural catheter with resultant failure to inject drug is a complication of lumbar epidural analgesia. Here, we report a case of kinking of epidural catheter 1 cm proximal to its tip after 20 days of insertion. It was inserted to a female for pain relief, suffering from carcinoma of the cervix. PMID- 14740600 TI - Cesarean section in a patient with Wolfe-Parkinson-White syndrome. AB - A 30 year old woman with abnormal fetal presentation was scheduled for emergency cesarean section. The patient had been a known case of Wolff-Parkinson-White syndrome, and appeared exceedingly anxious. She was given diazepam 5 mg and sufentanil 15 microgram as premedication following which routine and conventional general anesthesia was administered. The perioperative course was uneventful, and both the patient and the child were discharged fully well on the 5th postoperative day. PMID- 14740601 TI - Evaluation of i.v. verapamil effects on cardiovascular responses in normotensive patients during laryngoscopy. AB - Laryngoscopy and intubation are known to increase systolic blood pressure (SBP), diastolic blood pressure (DBP), mean arterial pressure (MAP), and heart rate (HR). In this study, we demonstrated that the injection of intravenous verapamil (0.1 mg/kg) prior to laryngoscopy can blunt the cardiovascular responses to laryngoscopy and intubation and result in stable hemodynamic profile in normotensive patients. PMID- 14740602 TI - Does LAD occlusion induce ischemic or hemodynamic modifications during MIDCAB? PMID- 14740604 TI - Global fund decides to promote use of generic drugs. PMID- 14740603 TI - WHO initiatives to increase access to treatment in developing countries. AB - In mid-2002, the World Health Organization (WHO) estimated that some six million people with HIV/AIDS in developing countries are currently in need of life sustaining antiretroviral (ARV) therapy, but that only 230,000 have access to these medicines, half of whom live in one country, Brazil. The WHO believes that, with a concerted international effort to expand access to HIV treatment and care, three million people could have access to ARVs by the end of 2005. A number of recent initiatives provide some useful tools toward reaching this goal. PMID- 14740605 TI - UK Commission concludes international patent laws hinder access to medicines in developing countries. AB - On 12 September 2002, the UK Commission on Intellectual Property Rights, an independent body established in May 2001 by the British government, released its report analyzing the impact of international agreements on patents. The report, Integrating Intellectual Property Rights and Developmental Policy, makes 55 recommendations "aimed at aligning [intellectual property] protection with the goal of reducing poverty". PMID- 14740606 TI - Thailand: successful challenge to invalid patent claim on antiretrovirals. AB - Thai people living with HIV/AIDS made legal history on 1 October 2002, when they won a lawsuit against the pharmaceutical giant Bristol Myers-Squibb (BMS). The plaintiffs, two people living with HIV/AIDS and a local NGO, the AIDS Access Foundation, lodged a complaint against BMS and Thailand's Department of Intellectual Property (DIP) in Thailand's Central Intellectual Property and International Trade Court (CIPITC). They claimed that the BMS patent registration for its buffered tablet formulation of the antiretroviral AIDS drug, dideoxy purine nucleoside (ddI, brand name Videx), was illegally amended in an attempt to claim a wider monopoly than the patent description justified. PMID- 14740607 TI - Mineralogical and chemical characterization of iron-, manganese-, and copper containing synthetic hydroxyapatites. AB - The National Aeronautics and Space Administration's (NASA) Advanced Life Support (ALS) Program is evaluating the use of Fe-, Mn-, and Cu-containing synthetic hydroxyapatite (SHA) as a slow release fertilizer for crops that might be grown on the International Space Station or at Lunar and Martian outposts. Separate Fe , Mn-, and Cu-containing SHA materials along with a transition-metal free SHA (pure-SHA) were synthesized using a precipitation method. Chemical and mineralogical analyses determined if and how Fe, Mn, and Cu were incorporated into the SHA structure. X-ray diffraction (XRD), Rietveld refinement, and transmission electron microscopy (TEM) confirmed that SHA materials with the apatite structure were produced. Chemical analyses indicated that the metal containing SHA materials were deficient in Ca relative to pure-SHA. The shift in the infrared PO4-mu 3 vibrations, smaller unit cell parameters, smaller particle size, and greater structural strain for Fe-, Mn-, and Cu-containing SHA compared with pure-SHA suggested that Fe, Mn, and Cu were incorporated into SHA structure. Rietveld analyses revealed that Fe, Mn, and Cu substituted into the Ca2 site of SHA. An Fe-rich phase was detected by TEM analyses and backscattered electron microscopy in the Fe-containing SHA material with the greatest Fe content. The substitution of metals into SHA suggests that metal-SHA materials are potential slow-release sources of micronutrients for plant uptake in addition to Ca and P. PMID- 14740608 TI - Clinical and economic impact of glatiramer acetate versus beta interferon therapy among patients with multiple sclerosis in a managed care population. AB - OBJECTIVE: To examine the outcomes of use of glatiramer acetate (GA) versus beta interferons-1a (intramuscular) (1A) and -1b (1B) in patients with multiple sclerosis (MS) in a managed care setting. METHODS: Data were obtained from a national retrospective claims database from January 1996 to June 2001. Patients were followed from the first prescription for immunomodulatory therapy until plan disenrollment or end of study time frame. The incidence of all relapses (defined as hospitalization for MS or ambulatory visit followed by use of systemic corticosteroids) as well as utilization and costs of MS-related care were examined for each group. Data were adjusted for variable follow-up using survival techniques. RESULTS: A total of 8,457 patients receiving immunomodulatory therapy were included in the study cohort; follow-up averaged 17.3 months. Three quarters of patients were female; the mean age was 42.2 years. The risk of relapse (defined as number of new cases) at one year was significantly increased for the beta interferons relative to GA (hazard rates: 1.15 and 1.51 for 1A and 1B, respectively, P<0.01). Mean (+/- SD) costs of care also were reduced among GA patients ($9,522 [+/- $9,706] versus $9,957 [+/- $9,083] and $10,185 [+/- $9,526] for 1A and 1B, respectively). These findings persisted in multivariate analyses, controlling for differences in demographic characteristics and propensity scores for immunomodulatory therapy. CONCLUSIONS: Glatiramer acetate is associated with reductions in the incidence of relapse and costs of care relative to the beta interferons among this large group of managed care patients with MS. PMID- 14740609 TI - Biotechnology, bioprocessing and bioengineering: who is doing what and why. September 25-26, 1998; Ohio University, USA. PMID- 14740610 TI - 9th World Conference on Lung Cancer. Tokyo, Japan. September 2000. PMID- 14740611 TI - Building biology across the Atlantic. PMID- 14740612 TI - Image-guided fine needle aspirate strategies for staging of lung cancer. AB - Image-guided transthoracic, bronchoscopic, and endoscopic ultrasound fine needle aspiration (FNA) greatly facilitates lung cancer staging by having the potential to precisely biopsy lung lesions and virtually all mediastinal lymph node stations. Imaging modalities alone, including chest x-ray, computed tomography (CT), magnetic resonance imaging, and positron emission tomography identify lesions suspicious for cancer but cannot make a tissue diagnosis. We describe an algorithm for the diagnosis and tumor-node-metastasis staging of lung cancer that uses procedures with the least invasiveness and cost with the highest diagnostic yields. For the anterior mediastinum, fluoroscopic-, ultrasound-, or CT-guided transthoracic FNA (which has a greater yield than bronchoscopy and is less invasive than mediastinoscopy) should be the primary technique for lymph node sampling. In the middle mediastinum, CT-guided transthoracic FNA is preferred for all nodal stations except subcarinal. Endoscopic ultrasound-guided FNA (EUS-FNA), which enables real-time biopsies within 5 cm of the esophagus, is preferred for sampling subcarinal and posterior mediastinal nodes because the yield is similar to CT-guided transthoracic FNA, with minimal risk of pneumothorax. The posterior mediastinum is also accessed by fluoroscopic- or CT-guided transthoracic FNA or video-assisted thoracic surgery. Sampling of the aorticopulmonary window depends on lymph node size; if the nodes are large enough to displace the aortic arch and pulmonary vein, then EUS-FNA is attempted, and if the nodes are not sufficiently enlarged, CT-guided transthoracic FNA should be performed prior to thoracoscopy or thoracotomy. PMID- 14740613 TI - Independent's days: independent colleges in the foundation of veterinary education in the United States. PMID- 14740614 TI - Early veterinarians in Dakota Territory, USA. PMID- 14740615 TI - Animal therapy over the ages: 10. Zoogenous therapy. Extracts, excretions, and secretions. PMID- 14740616 TI - The changing face of veterinary medicine: women in the profession. PMID- 14740617 TI - Medical oncology and palliative care: the intersection of end-of-life care. PMID- 14740618 TI - [The history of plague in the 18th century]. PMID- 14740619 TI - Reply to: Editorial--what residents need. PMID- 14740620 TI - [Medical issues in 16th century Hungarian calendars]. PMID- 14740627 TI - The falcon's stoop. PMID- 14740629 TI - [Bibliography of the Polish papers on forensic medicine, criminology and related fields published in 2002]. PMID- 14740630 TI - Complete small intestine mucosal recovery is obtainable in the treatment of celiac disease. PMID- 14740631 TI - Duodenal histology in patients with celiac disease after a gluten-free diet. PMID- 14740632 TI - Symptomatic small bowel intussusception: a surgical opportunity to diagnose adult celiac disease? PMID- 14740633 TI - Karl Kleist (1879-1960)- a pioneer of neuropsychiatry. AB - Karl Kleist (1879-1960) was instrumental in pioneering German neuropsychiatry and neuropsychology, including the description of frontal, constructional, limb kinetic (innervatory) and psychomotor apraxias, frontal akinesia and aspontaneity, as well as object and form blindness. Besides isolating episodic twilight states, involutional paranoia and symptomatic (especially influenza) psychoses, he was particularly involved in applying Wernicke's syndromatic and Kraepelin's prognostic and aetiological principles to classify "neurogenous" psychoses by refuting the assumption of mixed entities whenever possible. Thus, has phasophrenias denoted manic-depressive illness, unipolar affective disorders and marginal, i.e., atypical psychoses. The rather benign cycloid psychoses form the most prominent examples of the latter. Schizophrenias, on the other hand, were limited to poor long-term catamnestic outcomes. Kleist conceptualized the core group of schizophrenic illness as psychic system diseases-hence the origin of the term "systematic schizophrenias" within the Wernicke-Kleist-Leonhard School. Kleist was mainly influenced by Wernicke and his psychic reflex arc, but Ernst Mach's empiriocriticism, Theodor Meynert's cerebral connectionism, and associationism also shaped his outlook. Kleist's localization of cerebral functions by lesion analyses was indeed the best available at the time and continues to reveal insights to the interested reader. From his Frankfurt School, which may have been the last of a completely unified neuropsychiatry, came sound representatives of psychiatry, neurology and neurosurgery. His technical mastery and achievements seem indisputable, but his balancing acts during the Third Reich may today be questioned. Despite joining the National Socialist German Workers' Party (NSDAP) and the local Court of Genealogical Health (Erbgesundheitsgericht), Kleist was, however, one of the few German physicians who continued to treat Jewish patients, to employ Jewish colleagues and to voice evident criticism of the policies of "eugenics" and "euthanasia". This paper attempts to illuminate Kleist's biography and life's work in the relevant historical context. PMID- 14740634 TI - Samuel Wesley's "madness" of 1817-18. AB - In 1817 the musician Samuel Wesley was confined for a year in Blacklands House, a private lunatic asylum in London operated by Dr Alexander Robert Sutherland. Surviving documents enable aspects of Wesley's case to be reconstructed and indicate that his confinement was an enormous miscarriage of justice. PMID- 14740635 TI - Introduction: Adolf Meyer and his influence on British psychiatry. PMID- 14740637 TI - The clinical diagnosis and prognosis of mental disease. PMID- 14740636 TI - Remarks on Hecker's address. PMID- 14740638 TI - Cancer pharmacogenetics: polymorphisms, pathways and beyond. PMID- 14740639 TI - Mobile telephone use among Melbourne drivers: a preventable exposure to injury risk. PMID- 14740641 TI - Australian healthcare reform: in need of political courage and champions. PMID- 14740640 TI - Iatrogenic maxillary sinus exposure during maxillary posterior root-end surgery. PMID- 14740642 TI - Sinus bone grafting for oral antral fistulae. PMID- 14740643 TI - German Chemical Society (GDCh) biannual conference in Munich 2003. PMID- 14740644 TI - Three-dimensional distribution of bone density in the proximal humerus. AB - Bone quality of the proximal humerus is important for the surgical treatment of proximal humeral fractures and rotator cuff tears. However, very few studies have evaluated the areal bone mineral density (BMD) of the proximal humerus. The aim of this study was to analyze the volumetric BMD (vBMD) using peripheral quantitative-computed-tomography. Total, trabecular and cortical vBMD were determined separately for the proximal and distal half of the humeral head, the surgical neck and seven specific regions of interest. The greater tuberosity (GT) was divided into three regions, and the lesser tuberosity (LT) and articular surface (AS) were each divided into two regions. The proximal head showed a significantly higher trabecular (+ 46%) and cortical vBMD (+ 15%) than the distal one. The mean trabecular vBMD of AS was significantly higher (+ 80%), and the cortical vBMD was significantly lower (- 11%) than that of the tuberosities. In the proximal half of GT, trabecular vBMD was higher in the posterior than in the middle and anterior regions. Cortical vBMD was higher in middle region than in the anterior and posterior ones. In the distal half of GT, trabecular vBMD was significantly higher in the posterior than in the middle region, and cortical vBMD was significantly higher in the anterior than in the middle region. In one. These results point to bone sites that may provide stronger fixation for implants, reduce the risk of implant loosening, and therefore improve patient outcome. PMID- 14740645 TI - [Systemic scleroderma]. PMID- 14740646 TI - [Trends in the study on transdermal agents of Chinese medicinal herbs]. PMID- 14740647 TI - [Advances in the study on the effects of Chinese herbal drugs on apoptosis]. PMID- 14740648 TI - [Preliminary study on hypoglycemic activities of natural polysaccharides and their mechanism]. PMID- 14740649 TI - Development of surgical treatments for coronary artery disease. PMID- 14740650 TI - 25 years of European human spaceflight. PMID- 14740651 TI - Life in a spin: what has been learnt from space. PMID- 14740652 TI - The future is our business: how ESA prepares its technology. PMID- 14740653 TI - Rethinking and regrouping. PMID- 14740654 TI - Bumpy ride back to space. PMID- 14740655 TI - In space, CDs play a new tune. PMID- 14740656 TI - Habitation 2004 Conference abstracts, January 4-7, 2004, Orlando, FL. PMID- 14740657 TI - The main results of EVA medical support on the Mir Space Station. AB - The aim of this paper is to review the main results of medical support of 78 two person extravehicular activities (EVAs) which have been conducted in the Mir Space Program. Thirty-six male crewmembers participated in these EVAs. Maximum length of a space walk was equal to 7 h 14 min. The total duration of all space walks reached 717.1 man-hours. The maximum frequency of EVA's execution was 10 per year. Most of the EVAs (67) have been performed at mission elapsed time ranging from 31 to 180 days. The oxygen atmosphere of the Orlan space suit with a pressure of 40 kPa in combination with the normobaric cabin environment and a short (30 min) oxygen prebreathe protocol have minimized the risk of decompression sickness (DCS). There has been no incidence of DCS during performed EVAs. At the peak activity, metabolic rates and heart rates increased up to 9.9 13 kcal/min and 150-174 min-1, respectively. The medical problems have centred on feeling of moderate overcooling during a rest period in a shadow after the high physical loads, episodes with tachycardia accompanied by cardiac rhythm disorders at the moments of emotional stress, pains in the muscles and general fatigue after the end of a hard EVA. All of the EVAs have been completed safely. PMID- 14740658 TI - The commercial development of space: is an international regulatory framework needed? AB - The commercial space sector to date has failed to develop comprehensive regulations--"rules of the road"--for its international activities. Within the next 5 years, conflicts with respect to international trade in satellite sales and launch services could emerge, highlighting the need for such a regulatory framework. If the commercial space sector is to continue to develop, it is important to begin discussions now, before these conflicts become significant, on the elements of an appropriate international regulatory framework. The existing framework for space activities was developed when government, not commercial, space activities were dominant, or was adapted from regulations in other sectors such as terrestrial telecommunications. PMID- 14740659 TI - Blended control, predictor-corrector guidance algorithm: an enabling technology for Mars aerocapture. AB - A guidance scheme designed for coping with significant dispersion in the vehicle's state and atmospheric conditions is presented. In order to expand the flyable aerocapture envelope, control of the vehicle is realized through bank angle and angle-of-attack modulation. Thus, blended control of the vehicle is achieved, where the lateral and vertical motions of the vehicle are decoupled. The overall implementation approach is described, together with the guidance algorithm macrologic and structure. Results of guidance algorithm tests in the presence of various single and multiple off-nominal conditions are presented and discussed. PMID- 14740660 TI - Dynamics of space welding impact and corresponding safety welding study. AB - This study was undertaken in order to be sure that no hazard would exist from impingement of hot molten metal particle detachments upon an astronauts space suit during any future electron beam welding exercises or experiments. The conditions under which molten metal detachments might occur in a space welding environment were analyzed. The safety issue is important during welding with regards to potential molten metal detachments from the weld pool and cold filler wire during electron beam welding in space. Theoretical models were developed to predict the possibility and size of the molten metal detachment hazards during the electron beam welding exercises at low earth orbit. Some possible ways of obtaining molten metal drop detachments would include an impulse force, or bump, to the weld sample, cut surface, or filler wire. Theoretical models were determined for these detachment concerns from principles of impact and kinetic energies, surface tension, drop geometry, surface energies, and particle dynamics. A weld pool detachment parameter for specifying the conditions for metal weld pool detachment by impact was derived and correlated to the experimental results. The experimental results were for the most part consistent with the theoretical analysis and predictions. PMID- 14740661 TI - Mission to Mars using integrated propulsion concepts: considerations, opportunities, and strategies. AB - The aim of this paper is to evaluate the feasibility of a mission to Mars using the Integrated Propulsion Systems (IPS) which means to couple Nuclear-MPD-ISPU propulsion systems. In particular both mission analysis and propulsion aspects are analyzed together with technological aspects. Identifying possible mission scenarios will lead to the study of possible strategies for Mars Exploration and also of methods for reducing cost. As regard to IPS, the coupling between Nuclear Propulsion (Rubbia's engine) and Superconductive MPD propulsion is considered for the Earth-Mars trajectories: major emphasis is given to the advantages of such a system. The In Situ Resource Utilization (ISRU) concerns on-Mars operations; In Situ Propellant Utilization (ISPU) is foreseen particularly for LOX-CH4 engines for Mars Ascent Vehicles and this possibility is analyzed from a technological point of view. Tether Systems are also considered during interplanetary trajectories and as space elevators on Mars orbit. Finally strategic considerations associated to this mission are considered also. PMID- 14740662 TI - [Effects of ligustrazine on blood vessels and blood components]. PMID- 14740663 TI - [Advances in the study of Elsholtzia splendens]. PMID- 14740664 TI - [A review of the study on kudingcha]. PMID- 14740665 TI - [Advances in experimental study on pharmacodynamic effects of Eupolyphaga sinesis Walker]. PMID- 14740666 TI - [Advances in the study of Scolopendra--a kind of Chinese materia medica]. PMID- 14740667 TI - Deaths from eastern equine encephalitis reported in the south. PMID- 14740668 TI - Low compliance and arterial graft occlusion. AB - Early closure of arterial grafts is not fully understood. Patency loss in saphenous vein at 1 year is 12 to 30%. Synthetic grafts are worse. Stiff grafts may impede pulsatile flow and interfere with energy propagation. In vivo compliance (C, % radial change/mm Hg x 10(2) +/- SEM), derived from pressure and diameter measurements, was assessed in 38 arteries and 60 arterial grafts in a standardized canine femoral artery bypass model. Normal artery (C = 7.40 +/- 0.73) was more compliant than any graft (P < 0.01). At implantation, vein (C = 2.67 +/- 0.18) was more compliant than double velour Dacron (C = 1.86 +/- 0.16) or expanded polytetrafluoroethylene (C = 1.63 +/- 0.14) (P < 0.01). Vein graft patency at 2 weeks was 83%, Dacron 64% (P = 0.13), and PTFE 32% (P < 0.01). Vein compliance did not change with implantation. Vein patency was maintained, but Dacron fell to 29% and PTFE to 14% by 3 months. A definite correlation was shown between patency and compliance. It is suggested that more attention should be devoted to elastic properties in the design of arterial grafts. PMID- 14740669 TI - Noninvasive evaluation of asymptomatic carotid bruits. AB - Proper clinical management of a patient found to have an asymptomatic carotid bruit continues to be controversial, with wide differences of opinion concerning the advisability of angiography and surgery. A noninvasive cerebrovascular evaluation is described, which combines three separate but complimentary procedures: ocular pneumoplethysmography (OPPG), carotid audiofrequency analysis (CAA), and cerebral Doppler analysis. Such studies are particularly helpful in evaluating patients with an asymptomatic carotid bruit. The reliability and effectiveness of this approach was evaluated in a series of 165 consecutive patients with an asymptomatic bruit. Angiography was recommended in patients with positive results (42%) and safely withheld in those with negative findings (58%). There were two false-positive results and only one known false-negative interpretation. PMID- 14740670 TI - Ultrastructure of Hancock porcine valvular heterografts. Pre- and post implantation changes. AB - Scanning and transmission electron microscopic studies were made of: 1) 15 glutaraldehyde-treated porcine valvular heterografts that had been implanted in patients for periods of time ranging from 2 days to 76 months; 2) unimplanted, commercially processed porcine aortic valves; and 3) unprocessed porcine aortic valves. Unimplanted, commercially processed valves showed loss of endothelium and acid mucopolysaccharides. Short-term (< 2 months) post-implantation changes consisted of insudation of plasma proteins, penetration of erythrocytes into surface crevices, formation of a thin surface layer of fibrin, and deposition of macrophages, giant cells and a few platelets. Longer term (> 2 months) changes consisted of progressive disruption of collagen, erosion of valvular surfaces, formation of aggregates of platelets, and accumulation of lipid. The surfaces of the leaflets did not become covered with endothelium or with a fibrous sheath. It is concluded that progressive breakdown of collagen is a critical factor in determining the long-term durability of glutaraldehyde-treated porcine valvular heterografts. PMID- 14740671 TI - Systemic and pulmonary effects of nitroprusside during mitral valve replacement in patients with mitral regurgitation. AB - The effects of sodium nitroprusside on the systemic and pulmonary circulation were assessed in 17 patients with severe mitral regurgitation (MR) during mitral valve replacement. In nine patients, MR was related to rheumatic heart disease, in six resulted from acute myocardial infarction, and in two was due to subacute bacterial endocarditis. Nitroprusside was administered intraoperatively at an infusion rate of 10 to 65 mcg/min. During vasodilation therapy, forward stroke volume increased from 34 +/- 2.8 to 46 +/- 4.8 ml, while left ventricular filling pressure decreased from 28.7 +/- 1.9 to 20.6 < or = 1.8 mm Hg. There was a significant decrease in systemic vascular resistance (from 2379 +/- 204 to 1646 +/- 161 dynes.sec.cm-5) and mean arterial pressure (from 89.8 +/- 4 to 78.1 +/- 5 mm Hg). Similarly, a significant decrease was observed in both pulmonary vascular resistance (from 414 +/- 45 to 208 +/- 31 dynes.sec.cm-5) and mean pulmonary arterial pressure (from 42.5 +/- 2.5 to 29.7 +/- 2.5 mm Hg) with nitroprusside. In all patients there was a decrease in the magnitude of the peak V wave of the pulmonary capillary wedge pressure (from 45.2 +/- 3.9 to 26.1 +/- 3.1 mm Hg). These findings suggest that judicious intraoperative administration of nitroprusside improves right and left ventricular function in patients with MR undergoing mitral valve replacement, by increasing forward stroke volume while decreasing ventricular filling pressures and vascular resistance (preload and afterload). PMID- 14740672 TI - Intraoperative, intracardiac echocardiography during left ventriculomyotomy and myectomy for hypertrophic subaortic stenosis. AB - In 15 patients undergoing left ventriculomyotomy and myectomy for idiopathic hypertrophic subaortic stenosis (IHSS), the thickness of the interventricular septum was measured before and after muscle resection by means of a modified sternal-notch pediatric ultrasound transducer. Preoperative and intraoperative measurements by echogram were highly correlated (r = 0.92, P = 0.6), and accurately predicted the dimensions of the excised bar of muscle (r = 0.98, P = 0.6). The mean depth of the trough created at operation was 9.7 +/- 0.8 mm, and the mean thickness of the septum remaining was 14.3 +/- 0.9 mm. When studied 6 months postoperatively, all surviving patients were functionally improved, and none had a pressure gradient when studied in the resting state. However, two thirds of the patients had residual provocable gradients (decreased by 70% from those preoperatively). Intraoperative echogram measurements failed to correlate with the presence or extent of the postoperative provocable gradients. Intraoperative echocardiography provides imaging of dimensions otherwise unavailable, and may help define the characteristics of the myectomy that result in uniform symptomatic and hemodynamic benefit. PMID- 14740673 TI - Hemodynamic studies after De Vega's tricuspid annuloplasty. AB - Postoperative hemodynamic studies were conducted at rest and during exercise in 24 patients who, in addition to mitral and/or aortic valve surgery, had De Vega's tricuspid annuloplasty. To determine the degree of tricuspid insufficiency (TI), right atrial pressure (PRA) tracings, biplane right ventricular cineangiograms, and ultrasonic Doppler flow patterns were obtained. The TI improved in 14 patients, but still remained mild to moderate in 20 patients. In addition, a mild to-moderate tricuspid stenosis with pressure gradients from 2 to 7.8 mm Hg were found in 12 patients. There was no significant change in the mean PRA, with 8.0 +/- 4.5 mm Hg preoperatively and 7.5 +/- 3.5 mm Hg postoperatively at rest. During exercise, PRA rose to 17.0 +/- 6.5 mm Hg. The unsatisfactory results are also due to the persistent elevation of left atrial and pulmonary artery pressure after left heart surgery. De Vega's annuloplasty does not answer the tricuspid challenge, since the results are unpredictable. PMID- 14740674 TI - Hemodynamic evaluation of the Hancock-modified orifice bioprosthesis in the aortic position. AB - Porcine xenografts have had marginal hemodynamic function in the aortic position, particularly in the smaller sized valves. A new modified orifice heterograft prosthesis (MO) has been designed with the muscular leaflet of the porcine valve excised and replaced with the nonmuscular leaflet of a second valve, thus allowing 25 to 35% improvement in effective internal orifice area. In vitro studies on MO show clear hydraulic improvement over standard porcine prostheses and function comparable to that of low-profile metal prostheses. The MO has been implanted in 26 patients followed from 1 to 10 months with no prosthetic dysfunction or thromboemboli. There has been one perioperative and no late deaths. Intraoperative peak systolic gradients averaged 6.6 mm (0 to 18 mm) for 21 mm prostheses and 1.8 mm (0 to 10 mm) for 23 mm prostheses. These compare favorably with average gradients of 14.0 mm (5 to 27 mm) for standard 23 mm prostheses, 10.1 mm (0 to 30 mm) for 25 mm, and 4.0 mm (0 to 14 mm) for 27 mm prostheses. The MO valve combines the advantages of a bioprosthesis with the excellent hemodynamic performance necessary for the patient with a small aortic anulus. PMID- 14740675 TI - Mitral valve--the third time around. AB - Twenty-eight patients have undergone three operations on the mitral valve during 1958-1977. The first operation was closed mitral commissurotomy in 27 and open commissurotomy in one. After an average interval of 5.7 years, the 28 patients underwent a second operation. Operative procedures were: closed mitral commissurotomy in four, open commissurotomy in 14, and valve replacement in 10. After an average interval of 7.4 years, the third mitral valve operation was undertaken for congestive heart failure due to primary disease in 17 patients, prosthetic valve dysfunction in 10, and angina pectoris in one. At this time two patients were class II (New York Heart Association), 14 were class III, and 12 were class IV. Four died at 24 to 48 hours postoperatively, for a mortality rate of 14%. Analysis of risk factors revealed a significantly lower cardiac index (1.12 L/min/m2 +/- 0.35 SD) in those patients dying in the hospital compared to the cardiac index (2.73 L/min/m2 +/- 1.29 SD) of survivors. Deaths of patients undergoing a third operation on the mitral valve were related to poor ventricular function and not to previous surgery. PMID- 14740676 TI - Quantification of myocardial injury during coronary artery bypass graft. AB - Serial intraoperative myocardial-specific creatine-kinase (MB-CK) samples were obtained in 32 patients undergoing coronary artery bypass graft (CABG). Based upon their postoperative ECG and technetium pyrophosphate SCAN results, each patient was classified as either Group A (MI), B (normal), or C (equivocal). Peak MB-CK was reported for each group. The mean value for Group A (75 +/- 17 IU/L) is higher than for Group B (18 +/- 1 IU/L) or Group C (30 +/- IU/L). The concept of measuring and a formula for calculation of intraoperative myocardial injury are presented. The mean value for Group A (MI, 10,709 +/- 5885) is higher than either groups B (normal) or C (equivocal) by a test of proportionality (P < 0.001). Likewise, Group C (898 +/- 159) is higher than B (466 +/- 71) (P < 0.05). This index, while in need of further validation, corresponds closely to the clinical status of the 32 patients studied and should provide a means more refined than mortality or incidence of MI upon which to judge efficacy of any proposed means of operative myocardial preservation. PMID- 14740677 TI - Hypertension following coronary artery bypass graft surgery: comparison of hemodynamic responses to nitroprusside, phentolamine, and converting enzyme inhibitor. AB - Sustained systemic hypertension (mean arterial pressure 119 +/- 4 mm Hg) developed in 39 patients undergoing coronary artery bypass graft (CABG) surgery. In groups of patients with comparable mean arterial pressure, left ventricular filling pressure, heart rate, and cardiac output, treatment with sodium nitroprusside (NP) in 20 patients, phentolamine (PH) in 10 patients and nonapeptide converting enzyme inhibitor (CEI) in nine patients uniformly lowered mean arterial pressure by 26, 21, and 20%, respectively. Cardiac output increased during NP (5.7 +/- 0.1 to 6.4 +/- 0.3 L/min, P < 0.0005) and after CEI (5.7 +/- 0.4 to 7.4 +/- 0.5 L/min, P < 0.0005) but not after PH (5.7 +/- 0.3 to 5.9 +/- 0.3 L/min, P > 0.05). Left ventricular filling pressure decreased in all groups (P < 0.005), but heart rate increased during NP (91.3 +/- 2.4 to 98.4 +/- 2.2 beats/min, P < 0.0005 and after PH (95.5 +/- 2.1 to 99.2 +/- 3.0, P < 0.005), while it was unchanged after CEI P > 0.05). Of the nine patients with CABG hypertension who received CEI, seven exhibited a mean decrease in arterial pressure of 18.5 +/- 3.4%, while in the remaining two patients, mean arterial pressure was decreased only 3.4%. In the former, the pretreatment mean plasma renin activity (PRA) was 7.5 +/- 2.4 ng/ml/hr, while in the latter, mean PRA was 1.3 ng/ml/hr. PMID- 14740678 TI - Left ventricular aneurysmectomy in patients with single vs multivessel coronary artery disease. AB - To identify clinical syndromes benefiting from post-myocardial infarction (MI) left ventricular (LV) aneurysmectomy, 145 consecutively operated patients were followed a mean of 23 +/- 2 months postoperatively. Of the 145 patients, 49% had single-vessel (SV) coronary artery disease (CAD) (> or = 70% stenosis), 49% had multivessel (MV) CAD, and 2% (3 patients) had an unknown extent of CAD. Patients with SV-CAD and MV-CAD had similar distributions of age, sex, aneurysm location, LV end-diastolic pressure, LV ejection fraction, and scar size. In the SV-CAD group, the major operative indication was incapacitating heart failure (CHF), whereas in the MV-CAD group, the major operative indication was angina, often in combination with CHF. Coronary bypass grafting was done in 17/71 (24%) of SV-CAD and 64/71 (90%) of MV-CAD. Acute (30-day) postoperative survival was 89% (129/145) overall, and long-term survival 73%. Throughout the follow-up period, survival was similar in SV-CAD and MV-CAD. At follow-up, 89% of all survivors had CHF symptoms of functional Class II severity or less, and 97% of survivors had angina of Class II severity or less. Improvement in angina was most striking in the multivessel group whose angina had been more severe preoperatively. Thus, in patients undergoing LV aneurysmectomy and concomitant coronary revascularization: 1) SV-CAD is common; 2) SV-CAD has predominantly CHF preoperatively and usually relief of CHF postoperatively; 3) MV-CAD has both angina and CHF preoperatively, with symptomatic relief of both postoperatively; and 4) preoperative CAD extent does not influence survival. PMID- 14740679 TI - Potassium cardioplegia: early assessment by radionuclide ventriculography. AB - Left ventricular function was evaluated by single pass Tc-99m radionuclide ventriculography when potassium cardioplegia was combined with hypothermia. In 35 patients undergoing myocardial revascularization (3 CABG/patient) in which potassium cardioplegia at 4 degrees C was used, no patient developed a myocardial infarction either by electrocardiogram or Tc-99m pyrophosphate imaging in the postoperative period. In 22 patients, aortic cross-clamp time was greater than 60 min, and the ejection fraction by the single pass radionuclide technique was 50% preoperatively and 53% postoperatively (NS). Wall motion in the single RAO view was not worse postoperatively. No patient required any inotropic agents in the immediate postoperative period. It appears that no significant ventricular impairment occurred in the immediate postoperative period (48 to 72 hours) when potassium cardioplegia combined with hypothermia was used for a 60-minute period. PMID- 14740680 TI - Changes in left ventricular segmental wall motion following randomization to medicine or surgery in patients with unstable angina. AB - Twenty-six patients with unstable angina pectoris had biplane left ventricular (LV) angiograms and coronary arteriograms (CAGs) initially and at a median of 1 year following randomization to medical (15 patients) or surgical 11 patients) therapy. Left ventricular segmental wall motion was analyzed by a digital computer yielding segmental ejection fraction for 10 zones along the LV perimeter. Baseline and follow-up CAGs were analyzed simultaneously by one observer, and changes in LV segmental coronary perfusion were estimated. Left ventricular angiograms were analyzed separately from CAGs and independently of knowledge of changes in estimated segmental coronary perfusion. Left ventricular segmental wall motion was more frequently improved in surgical patients than in medical patients. Furthermore, in surgical patients there was a significant correlation between changes in LV segmental wall motion and perfusion to LV segments supplied by the left anterior descending coronary artery, whereas no such correlation for any segment was found in the medical group. Thus, this prospective randomized study suggests that, in patients with unstable angina, coronary revascularization may significantly improve LV segmental wall motion compared to medically treated patients. In the surgical group, improvement in LV wall motion relates to improvement in coronary perfusion to the segments supplied by the left anterior descending artery. PMID- 14740681 TI - Management of acute myocardial ischemia with intraaortic balloon pumping and coronary bypass surgery. AB - Ninety-three patients with unstable angina pectoris resistant to maximum medical therapy were treated with intraaortic balloon pumping (IABP), early angiography, and myocardial revascularization. Sixty patients had no recent infarction: 42 with typical angina (ST depression > 0.1 mV) and 18 with variant angina (ST elevation > 0.2 mV). Thirty-three patients were post infarction (< 10 days), 25 with typical angina and eight with variant angina. IABP interrupted ischemic attacks in 75/93 (81%) and reduced frequency and intensity of attacks in the remaining patients. Mortality from revascularization was 5/93 (5.4%) with two deaths in the preinfarction group (3.3%) and three deaths in the postinfarction group (9.1%). Incidence of perioperative myocardial infarction was 2.2%. With an average follow-up of 38 months there have been four late deaths (4.5%), three secondary to myocardial infarction, and only one other symptomatic infarction. After operation, 93% have no significant angina, and 70% are fully active. When IABP is used to control recurrent ischemia, patients with refractory angina pectoris can be safely revascularized and have a favorable long-term prognosis. PMID- 14740682 TI - Blalock-taussig anastomosis: the preferred shunt in infants and newborns. AB - The Blalock-Taussig anastomosis (B-T shunt) has been regarded as an unsatisfactory procedure for infants and newborns with severe pulmonary oligemia. With use of microsurgical technique, B-T shunts were constructed in 11 infants under 6 months of age, six of whom were less than 1 month old (2 to 30 days). Weights ranged from 2.4 to 5.6 kg. Diagnoses included transposition of the great arteries (4), tetralogy of Fallot (4), pulmonary arterial atresia (2), and Taussig-Bing syndrome (1). There was no operative mortality; perioperative morbidity included paralysis of the diaphragm (2), Horner's syndrome (1), superficial wound dehiscence (1), congestive heart failure (1), and prolonged respiratory failure requiring ventilation assistance for more than 5 days (2). There has been one late death in the group at 2 months post-shunt. All surviving patients have had satisfactory palliation of cyanosis and hypoxemia. In view of the pulmonary artery scarring and kinking that follows the Waterston and Potts shunts, it is concluded that the Blalock-Taussig shunt is a reliable and preferable technique for palliation of pulmonary oligemia in infants and newborns. PMID- 14740683 TI - Late results of the subclavian flap procedure in infants with coarctation of the thoracic aorta. AB - Persistent stenosis or recoarctation occurs in 16 to 33% of infants who undergo repair of coarctation of the thoracic aorta by use of end-to-end anastomosis. This report describes the results in 12 infants of repair of coarctation of the aorta using a left subclavian artery flap procedure. Cardiac catheterization and cineangiography performed in four patients from 4 to 66 months following repair showed a 10 mm systolic gradient in one patient and no evidence of significant stenosis in any patient. The eight other patients have had recent sphygmomanometer pressures recorded from 1 to 29 months after repair. Four patients had no gradient while three patients had a systolic pressure gradient of 8, 10, and 12 mm Hg. Results suggest a smaller incidence of persistent stenosis or recoarctation following subclavian flap procedure than following end-to-end anastomosis. The subclavian flap operation provides an adequate aortic lumen with good growth potential. It is recommended as the operation of choice for infants with the most common form of coarctation of the aorta. PMID- 14740684 TI - Physiological approach to surgery for tricuspid atresia. AB - A modification of Fontan's approach to the surgical management of tricuspid atresia was employed in nine patients. A valved conduit was used to create continuity between the right atrium and the surgically modified right ventricular chamber. Six patients had had previous shunting procedures. Preoperative studies revealed various degrees of hypoplasia of the right ventricle beneath relatively normal pulmonary valves. At surgery, the atrial septal and ventricular septal defects were closed, the right ventricular chamber was enlarged, and right atrial right ventricular continuity was established with a valved conduit. There was one operative death. The eight survivors have been clinically well as long as 3 1/2 years following surgery. Postoperative cardiac catheterization was performed in five patients. Ventricularized pressure tracings were recorded in the right ventricular chambers and the morphology of the atrial and pulmonary artery pressure curves were normal. In all instances, angiography demonstrated significant enlargement of a pulsatile right ventricular chamber. Thus, in many patients with tricuspid atresia, establishment of a four-chambered, four-valved heart is feasible, both from an anatomic and physiologic standpoint. PMID- 14740685 TI - Ebstein's anomaly of the left-sided tricuspid valve: pathological anatomy of the valvular malformation. AB - The tricuspid valve is left-sided in congenitally corrected transposition of the great arteries and other cardiac malformations in which the ventricles are inverted. In an anatomical study of 20 hearts with ventricular inversion, nine were found to have Ebstein's anomaly of the left-sided tricuspid valve. These included eight with congenitally corrected transposition of the great arteries and one with a double outlet inverted right ventricle. Pathoanatomically left sided Ebstein's anomaly differed from the classic right-sided variety in several respects: 1) the atrioventricular sulcus circumference was not increased; 2) the anterior leaflet of the valve was frequently cleft; 3) the anterior leaflet may interfere with the ventricular outflow; 4) the ventricular cavity receiving the malformed tricuspid valve was small, rather than dilated--and in congenitally corrected transposition of the great arteries, this is the systemic ventricle connected to the aorta. Significant hemodynamic dysfunction of the left-sided tricuspid valve has been reported in a high proportion of cases of congenitally corrected transposition of the great arteries, and surgical repair of the valve may be necessary. Our study suggests that surgical procedures used in classic right-sided Ebstein's anomaly, such as valve plication and annuloplasty or valve replacement with a high profile prosthesis, are not suited to the pathoanatomy of left-sided Ebstein's anomaly. PMID- 14740686 TI - Myocardial revascularization in patients with multivessel coronary artery disease and minimal angina pectoris. AB - Mortality risk in coronary artery disease (CAD) is more closely related to angiographic findings of multiple coronary artery obstructions and left ventricular asynergy than to the severity of angina pectoris, the major symptom of CAD. Since coronary revascularization surgery is most frequently performed to relieve chest pain, there are few reports evaluating the results of coronary artery bypass surgery in patients with minimal or no angina pectoris but with anatomically severe disease. From July, 1970, through December, 1976, 844 patients had coronary artery bypass surgery performed at the Peter Bent Brigham Hospital for chronic or unstable angina pectoris. Twenty patients (2.3%) were operated on because of severe coronary obstruction but who had minimal or no angina. Fourteen patients underwent coronary arteriography because of a positive exercise tolerance test, and six because of a prior myocardial infarction. All but one patient had multivessel CAD, and four patients had significant left main coronary lesions. There was no operative mortality. One late death occurred 5 years postoperatively, for a 5.0% cumulative mortality. Average follow-up has been 34 months (range, 19 to 80 months). Of 12 patients with both pre- and postoperative exercise tests, eight have reverted to normal, and four show a less ischemic response to exercise. Coronary revascularization may have a beneficial effect on the patient with "asymptomatic" but anatomically severe CAD. PMID- 14740687 TI - Long-term patency of saphenous veins for coronary bypass grafting. AB - The patency of 871 aortocoronary saphenous vein grafts in 438 patients was evaluated for a mean (+/- SEM) of 12.1 +/- 0.3 months after operation. Second studies were performed in 62 patients (134 grafts) at a mean of 42 +/- 1.3 months postoperatively (range, 13 to 61 months). The 1-year patency rate was 79.4% and increased from 76% for 546 grafts placed in 1970-1973 (Period I) to 85% for 325 grafts in 1974-1976 (period II) (P = 0.001). Overall patency was highest (82%) for grafts to the anterior descending system and lowest (76%) for the right system. These patency rates also increased between periods I and II from 79 to 88% (P < 0.05) for the anterior descending and 72 to 83% (P < 0.05) for the right system. For 47 right coronary arteries having associated distal manual endarterectomy, the graft patency rate was 62% vs 78% for those not endarterectomized (P < 0.05). Of 116 grafts patent at the initial study, five were occluded at the second study, a late failure rate of 3.7% and a mean annual attrition rate of 1.7% between 12 and 42 months after operation. High-grade (> 50%) stenoses were present in three of the five grafts at initial study. Development of late stenosis was observed in two grafts (1.7%). These data document improved 1-year patency rates for grafts in period II and a low incidence of late failure and development of late stenosis for up to 61 months following operation. Continued use of saphenous vein grafts to all three coronary arterial systems appears justified. PMID- 14740688 TI - Progression of obstructive coronary artery disease 5 to 7 years after aortocoronary bypass surgery. AB - Changes in the native coronary circulation were evaluated 5 to 7 years (72 +/- 10 months) after aortocoronary bypass surgery in 108 patients; 104 had had previous angiographic controls between 6 and 18 months following surgery. Coronary artery narrowings were graded on a scale of 0 to 6, 0 being normal, grade 1 and 2, < or = 50%; grade 3, 51-75%; grade 4, 76-90%; grade 5, 91-99% narrowing and grade 6 total occlusion. Progression of disease was significant when a preexisting stenosis increased at least 1 grade or a new lesion > 50% developed. Nongrafted vessels were compared to arteries with patent or occluded grafts. During the first year following surgery, the rate of progression was much higher in grafted arteries with patent or occluded grafts than in nongrafted arteries 157 and 53% vs 9.5% respectively, P < 0.001). During the subsequent period of approximately 5 years, however, grafted arteries did not change, whereas the rate of progression in nongrafted arteries increased from 9.5 to 46%. At 5 to 7 years after surgery, grade 1 and 2 (< or = 50%) narrowings progressed slightly more frequently in grafted arteries because of surgically related events. However, for narrowings > 50% and for new lesions, there was no difference in the rate of progression between grafted and nongrafted vessels. In conclusion, the rate of progression of atherosclerosis in nongrafted coronary arteries is important. At 5 to 7 years after surgery, there is little or no difference in the severity of the disease in the proximal segments of grafted and nongrafted coronary arteries. PMID- 14740689 TI - Cardioplegia and myocardial preservation during cardiopulmonary bypass. AB - A standard experimental protocol was developed to explore the role of hypothermia and potassium cardioplegia in myocardial preservation during 120 minutes of ischemic arrest followed by 30 minutes of reperfusion. Seven different experimental groups of six animals each were evaluated using an in-vivo pig heart preparation. Hypothermic arrest without cardioplegia and cardioplegic arrest at normothermia were each compared to hypothermic cardioplegia. In addition, the use of an asanguineous hypothermic coronary perfusate without cardioplegia was compared to both multidose cardioplegia and single-dose cardioplegia followed by the same asanguineous perfusate. The parameters measured included: myocardial contractility and compliance, myocardial blood flow, endocardial/epicardial blood flow ratio, and electron microscopic studies. Myocardial preservation was inadequate with hypothermic arrest alone (without cardioplegia; and with cardioplegia at normothermia. In both experimental groups, myocardial contractility and compliance were so depressed that the) could not be accurately measured following ischemia and reperfusion while coronary blood flow remained significantly elevated. Preservation was improved but still inadequate following myocardial washout with a normokalemic or hypokalemic perfusate and following single dose cardioplegia plus myocardial washout. In the latter four groups, contractility ranged from 42 to 78% of control, and there was a decrease in compliance of 16 to 78%. Adequate preservation was found only after hypothermia and multidose potassium (35 mEq/L) cardioplegia. In this group, contractility was 129 +/- 13% of control and compliance increased by 21 +/- 24% compared to that of the control. PMID- 14740690 TI - Cardioplegic solutions for myocardial preservation: analysis of hypothermic arrest, potassium arrest, and procaine arrest. AB - Separate effects of perfusion hypothermia, potassium arrest, and procaine arrest were studied in 150 hearts using the isolated rat heart preparation. Aortic flow rate (AFR), coronary flow rate (CFR), and heart rate (HR) were measured before and after ischemic periods of 90 to 300 min. Prior to the ischemic period, the aortic root was infused with Krebs-Henseleit buffer (KHB), buffer with 30 mEq KCl/L (KHB + K), or buffer with 0.2% procaine (KHB + P), at 15 degrees or 5 degrees C. During the ischemic period the hearts were maintained at 15 degrees or 5 degrees C in a hypothermic chamber. The three solutions had similar recoveries of AFR at 15 degrees and 5 degrees C following ischemic periods of 90, 180, and 240 min. The KHB + K and KHB + P had better recoveries of AFR than KHB at 5 degrees C and 300 min of ischemia. The KHB + K and KHB + P also caused more rapid arrest and a higher incidence of spontaneous recovery of sinus rhythm. Arrest at 5 degrees C was equal to or superior to arrest at 15 degrees C. Perfusion hypothermia is the main component of cold cardioplegia. The addition of procaine or potassium results in increased functional recovery at extended ischemic times at 5 degrees C, more rapid arrest, and better electrical recovery. PMID- 14740691 TI - Effects of verapamil on myocardial tolerance to ischemic arrest: comparison to potassium arrest. AB - This study evaluates the metabolic and physiologic effects of Verapamil on isolated, perfused rat hearts subjected to 1 hour of global ischemia. Hearts were perfused with a modified Krebs-Henseleit bicarbonate buffer with glucose. During working heart perfusion, mean aortic systolic pressure, heart rate, and aortic and coronary flows were measured. Cardiac output and minute work were calculated. In three groups of eight hearts, 60-minute global ischemia was followed by a 40 minute reperfusion to assess physiologic recovery. Three groups of 11 hearts were assayed for myocardial high-energy phosphates after ischemia. Untreated hearts recovered less than 15% of function and lost 50% of their high-energy phosphate stores. Hearts treated with Verapamil (2 microg/cc) prior to ischemia recovered greater than 85% of function and retained normal adenosine triphosphate (ATP) and 40% decreased creatine phosphate (CP) levels. The metabolic and physiologic effects of Verapamil lasted 20 minutes longer than potassium, possibly related to membrane binding and dissociation of Verapamil. Verapamil was shown to have cardioplegic and protective effects, both probably secondary to inhibition of calcium flux across the sarcolemmal membrane. PMID- 14740692 TI - Clinical experience with intraaortic balloon pump support in 728 patients. AB - From November, 1968, through December, 1976, intraaortic balloon pump (IABP) circulatory support was used 747 times in 728 patients. Overall in-hospital survival was 413/747 (56.9%). Survival with IABP since 1974 has been 65% (286/440) compared to 24% (18/75) in 1968-1971 (P < 0.001). Two practical changes have been associated with this improvement: 1) broadened indications for, and earlier insertion of, IABP; and 2) more aggressive surgical treatment of the primary cardiac lesion. Cardiogenic shock or chronic ischemic ventricular failure accounted for 78.7% (59/75) of IABP in 1968-1971; during 1974-1976 only 26.4% (116/440) of IABP-supported patients were in these categories (P < 0.001). Since 1974, 58.2% (153/263) of IABP patients had an operative procedure following IABP insertion compared to 37.7% (23/611 prior to 1971 (P < 0.01). The major complication rate has remained unchanged at 8.5%. IABP-related mortality rate was 0.8% (6/728). Earlier, more liberal, use of IABP hemodynamic support has led to increased survival in a large number of patients with complications of acute myocardial ischemia. The low mortality associated with its use vindicates earlier institution of IABP support in cardiogenic shock. PMID- 14740693 TI - Prolonged mechanical support of the left ventricle. AB - An interdisciplinary group has developed a left ventricular assist pump system composed of a modified sac type pump, a pneumatic power unit, and a synchronizer. The pump fills from the left ventricle and discharges into the aorta. The system was employed for left ventricular assistance in a series of 12 normal calves, with an average pumping period of 70 +/- 8 days. The system was then evaluated in a series of calves in whom profound left ventricular failure had been produced. These studies indicate that the assist pump is effective in supporting the circulation and completely unloading the left ventricle. The assist system has now been employed in four patients who could not be weaned from cardiopulmonary bypass following cardiac valve replacement. The assist pump supported the circulation in three instances. In one patient, the assist pump was employed for 8 days until left ventricular function had improved sufficiently to permit pump removal; the patient was subsequently discharged from the hospital. PMID- 14740694 TI - Perioperative infarction: effects of cardiopulmonary bypass on collateral circulation in an acute canine model. AB - Significant intracoronary collateralization [circumflex (Cx) to anterior descending (AD)] can be established acutely by selective hypertensive perfusion of the circumflex (through left main) after acute occlusion of the AD in working canine hearts. Intracoronary collateral flow can be estimated by a collateral circulation index (CCI) = distal coronary diastolic pressure x 100/aortic diastolic pressure. The effects of 60 to 90 min of normothermic, total cardiopulmonary bypass (TCPBP) on such collaterals were evaluated in four dogs with CCI > 80%, and compared to six control dogs with CCI < 80% and without TCPBP. Three hours after AD ligation, control dogs had minimal myocardial injury despite significantly lower CCI. In contrast, the four study dogs placed on TCPBP experienced an abrupt fall of the CCI to 30% (P < 0.03), severe ischemia, and infarction in each; two became pump dependent. Thus, ventricular pressure work seems to be an important determinant of flow through collateral channels. These results suggest that preservation techniques for myocardium supplied by collaterals during coronary surgery may require considerations other than intermittent perfusion via existing coronary inflow. PMID- 14740695 TI - Hemodilution causes decreased compliance in puppies. AB - Cardiopulmonary bypass and hemodilution in the newborn have been associated with increased myocardial edema, which may be due to immature connective tissue formation. Five adult and four puppy dogs were placed on bypass while compliance and ventricular function (intraventricular balloon) were measured during normohemoconcentration (NH) (hematocrit 45.1, osmolality 307) and hemodilution with normal saline (hematocrit 24.7, osmolality 307). Compared with NH, the adult group showed no change in compliance or function after 90 min of hemodilution. The puppy group showed a marked decrease in compliance with hemodilution compared to NH (P < 0.001) without change in ventricular function when peak systolic pressure was plotted against end-diastolic volume. Electron micrographs confirmed greater edema formation in the puppy group than the adult group with hemodilution. These changes demonstrate that the newborn heart is more sensitive to edema formation than the adult during hemodilution and that filling pressures do not necessarily reflect ventricular performance during the early post perfusion period when compliance is decreased. PMID- 14740696 TI - Hemodynamic characteristics of the composite strut ball valve prostheses (Starr Edwards track valves) in patients on anticoagulants. AB - The hemodynamic characteristics of the composite strut ball valve prostheses (Starr-Edwards Model 2400 aortic track valve and Model 6400 mitral track valve) were evaluated in anticoagulated patients. The peak systolic gradient (mean +/- SE) for the aortic valve in 16 patients was 18.1 +/- 3.9 mm Hg; the mean gradient was 17.0 +/- 2.7 mm Hg; the prosthetic valve area was 1.7 +/- 0.1 cm2; and the valve area index was 1.0 +/- 0.1 cm2/m2. In the 19 patients with the mitral valve, the mean diastolic gradient (mean +/- SE) was 5.2 +/- 0.7 mm Hg. The prosthetic valve area was 2.0 +/- 0.1 cm2, and the valve area index was 1.3 +/- 0.1 cm2/m2. These hemodynamic parameters are helpful in evaluating the catheterization findings in patients with composite strut prosthetic valves who are suspected of having prosthesis dysfunction. They also demonstrate the satisfactory in vivo hemodynamics in anticoagulated patients with these prostheses. PMID- 14740697 TI - Pulsatile flow characteristics of aortic valve porcine heterografts in man. AB - Pulsatile flow characteristics in the ascending aorta were examined in 14 patients 12 months after aortic valve replacement with the glutaraldehyde-fixed porcine aortic heterograft. Analysis of the velocity signals permitted measurement of peak velocity, maximal acceleration, stroke volume, and ejection time in addition to the standard measurement of systolic aortic valvar gradient. Angiographic techniques permitted visualization of a pattern of blood flow in the ascending aorta. Valvar aortic gradients were minimal, and, when the phasic ascending aortic blood flow velocity parameters were compared to those in a group of eight patients with normal aortic valves, these characteristics of flow were nearly indistinguishable. This remarkable similarity to the flow characteristics of normal valves may be important to long-term performance of a prosthetic aortic valve. Our results suggest that the glutaraldehyde-fixed porcine heterograft warrants serious consideration when selecting a substitute aortic valve for man. PMID- 14740698 TI - Effect of surgery on ventricular tachyarrhythmias associated with coronary arterial occlusive disease. AB - We studied 51 patients with coronary artery disease over a 32-month period to determine the effect of coronary revascularization or ventricular resection on ventricular tachyarrhythmias. In these patients, whose major indications for operation were tachyarrhythmias, operative mortality was 18% (9/51). Myocardial infarction could not be documented in 16 patients, including four resuscitated from an arrest; 15 underwent coronary grafting, and one had grafting and mitral valve replacement, with one hospital death. Premature ventricular contractions (PVCs) persist in four of the 15 survivors; all those who had an arrest are free of PVCs. The other 35 patients had documented infarction, associated with an arrest in 19. Grafting only was required in 18, with no hospital mortality and two late deaths (congestive failure and ventricular tachyarrhythmia at 7 months, ventricular tachyarrhythmia at 5 months); eight of 16 survivors have PVCs. Acute (< 3 weeks) infarct resection was performed in six patients with three hospital deaths, no late deaths; one survivor has PVCs. Chronic (> 3 weeks) aneurysm resection in four patients (with bypass grafting in three) had no operative mortality, one late death (5 months) from ventricular tachyarrhythmias, and one survivor continues with PVCs. The remaining seven patients, all in cardiogenic shock, required more than two procedures (bypass grafting, LV resection, VSD closure, mitral valve replacement), with five hospital deaths; the two survivors still have PVCs. Thus, ventricular tachyarrhythmia secondary to coronary artery disease is more likely to resolve postoperatively in patients without myocardial infarction, but its postoperative persistence remains substantial and may warrant special efforts to locate and ablate ectopic foci and/or re-entry pathways. PMID- 14740699 TI - Aortocoronary saphenous vein bypass graft changes 5 to 7 years after surgery. AB - Control angiographic studies were carried out 54 to 88 months after aortocoronary saphenous vein bypass surgery (70 +/- 10 months) in 100 unselected patients who were known to have at least one patent graft 6 to 18 months following the operation. Only 17 of the 159 grafts were found occluded during this interval of 5 years, giving an average yearly attrition rate of 2.1% after the first year. The mean yearly attrition rate was 0.7% in our second series of near consecutive patients operated on after 2 years of experience and modifications of surgical techniques, and it compares favorably with the 2.4% yearly attrition rate found in our first series of patients (P < 0.05). Of the 37 grafts having localized stenoses at 1 year, eight became occluded (21.6%), and four became worse. Grafts with localized stenoses were more prone to late occlusion, whereas no late occlusion was observed in normally appearing grafts at 1 year. Diffuse graft narrowing noted during the first year showed no further change during the subsequent 5 years, and it did not appear to lead to late occlusion. Finally, in 14 grafts, localized narrowing developed after the first year, at a yearly rate of 1.5%. Atherosclerosis appears to be the most likely etiology, being found in two of these instances at reoperation. PMID- 14740700 TI - Left ventricular function during intra-aortic balloon pumping assessed by multigated cardiac blood pool imaging. AB - The effectiveness of intra-aortic balloon pumping in augmenting ventricular performance during acute myocardial ischemic syndromes was determined by noninvasive multigated cardiac blood pool imaging. Nine patients with unstable angina refractory to medical therapy and nine patients with acute myocardial infarction complicated by congestive heart failure were studied on and off intra aortic balloon pumping (IABP). End-diastolic and end-systolic volumes were significantly reduced by IABP in both unstable angina and myocardial infarction. Analysis of ventricular wall motion demonstrated improved segmental contraction during IABP in all seven unstable angina patients with regional wall motion abnormalities. No significant improvement in regional contraction was observed in the nine patients with acute myocardial infarction. During IABP, percent ventricular asynergy decreased significantly (28.2%; P < 0.01) among patients with unstable angina but demonstrated no significant fall (4.4%; P = NS) for patients with myocardial infarction. Thus, IABP enhances contraction of ischemic but not infarcted myocardium and effectively improves ventricular function by reducing end-diastolic and end-systolic volumes. PMID- 14740701 TI - Effects of calcium administration during post-ischemic reperfusion on myocardial contractility, stiffness, edema, and ultrastructure. AB - Previous investigators have suggested that calcium may play a role in the pathogenesis of myocardial cell damage following ischemia and reperfusion. Twenty six in-situ blood perfused isovolumic canine preparations were divided into four groups. Group I dogs were maintained normocalcemic during 45 min of reperfusion following 45 min of hypothermic (27 degrees C) ischemic arrest; Group II dogs received CaCl2 (7 mg/kg) after 15 min of reperfusion; Group III dogs received citrate solution (0.8 ml/kg citrate-phosphate-dextrose [CPD]) after 15 min of reperfusion; Group IV dogs received 7 mg/kg of CaCl2 at 5 min after receiving the same citrate dose as Group III after 15 min of reperfusion. In Group II hearts, calcium improved the left ventricular contractility (P < 0.05 vs Group I) without causing additional cellular or subcellular injury. Calcium also appeared to increase myocardial stiffness (alpha(n)) compared to Group I hearts (P < 0.01). In Group III hearts, citrate reduced contractility (P < 0.01 vs Group I) and increased myocardial edema (P < 0.005 vs Group I) without any apparent improvement in cellular or subcellular preservation. In Group IV hearts, calcium reversed the depression of contractility caused by citrate, resulted in no additional morphologic injury, increased myocardial stiffness compared to Group I or Group III (P < 0.005), and minimized myocardial edema (P < 0.005 vs Group I or III). These results suggest that calcium administered after 15 min of reperfusion improves the depression of contractility that follows hypothermic ischemic arrest without causing additional myocardial damage. PMID- 14740702 TI - Six-year clinical and angiographic follow-up of patients with previously documented complete revascularization. AB - Relatively little information is available concerning the late clinical and angiographic status of patients with initially successful coronary bypass surgery. From 72 patients who had angiography 1 year after bypass surgery, we restudied at 6 years 19 patients with complete revascularization. At 1 year, 14 patients were asymptomatic and five had minimal anginal symptoms. Five years later, eight patients had redeveloped angina, and 11 retained their initial postoperative status. Overall graft patency at 6 years was 86%; 52% of the patients had atherosclerotic progression to > or = 70% luminal narrowing in a major unbypassed vessel or in a major vessel distal to bypass. The patients with unchanged symptoms all had patent grafts, while 11 of the 15 (73%) grafts were patent in patients with symptomatic deterioration (NS). However, progression of coronary disease occurred in seven of eight patients (88%) with worsened symptoms, as opposed to three of 11 patients with unchanged symptoms (P < 0.05). We conclude that late symptomatic deterioration following coronary bypass surgery is common, and that it usually reflects progression of coronary artery disease. PMID- 14740703 TI - Advantages of intermittent blood cardioplegia over intermittent ischemia during prolonged hypothermic aortic clamping. AB - Ten dogs underwent 2 hours of hypothermic (22 degrees C) aortic clamping. Arrest was produced by ischemia in five dogs, with 150 ml of 22 degrees C unmodified blood reperfused every 20 minutes. Five others underwent cardioplegic arrest (500 cc pH 7.8 blood containing 30 mEq/L KCl, 0.6 mEq/L Ca++) with 150 ml of blood cardioplegia solution replenished each 20 minutes. Seven additional dogs underwent 4 hours of continuous coronary perfusion without ischemia. Continuous coronary perfusion did not change myocardial ATP or water content, but reduced left ventricular contractility (maximum positive dP/dt, peak systolic pressure) 16% (P < 0.05) and decreased left ventricular compliance moderately (50%). Hypothermic ischemic arrest reduced myocardial ATP 39% (P < 0.05), raised myocardial water 3.2% (P < 0.05), reduced compliance 83% (P < 0.05), and depressed left ventricular performance 64% (P < 0.05), despite intermittent reinfusion of unmodified blood each 20 minutes. In contrast, blood cardioplegia reduced postischemic left ventricular compliance only slightly 17%, P < 0.05) and resulted in normal postischemic ATP, water, and contractility. PMID- 14740704 TI - Long-term hemodynamic and electrocardiographic assessment following operative repair of tetralogy of Fallot. AB - To evaluate the long-term results of operative repair of tetralogy of Fallot, the cardiovascular, clinical, and electrocardiographic status was evaluated in 21 patients who had undergone operation 5 to 19 years previously. All patients were asymptomatic. Major functional abnormalities detected were enlarged right ventricular dimension in 15 of 17 patients, and elevated right ventricular systolic pressure during maximal exercise (mean, 69 +/- 5 mm Hg). Nevertheless, all patients had normal or nearly normal cardiovascular responses to maximal exercise as measured by cardiac output, maximal oxygen consumption, and left ventricular ejection fraction. Significant ventricular arrhythmias were detected during 24-hour ambulatory monitoring in 14 of 21 patients. Multiform ventricular premature beats (VPBs) were seen in 13 of 14 patients, couplets in four patients, greater than 30 VPBs for at least 1 hour in three, 4-complex ventricular tachycardia in one, and repeated R on T episodes in one. Although clinical and hemodynamic function was excellent in most patients, close follow-up is still necessary to determine the significance of the high-grade ventricular arrhythmias detected and if late manifestations of cardiovascular deterioration will occur. PMID- 14740705 TI - Cardiac metabolism and performance following cold potassium cardioplegia. AB - The ability of cold potassium cardioplegia (CPC) to preserve cardiac metabolism and performance was evaluated in 68 patients undergoing anoxic arrest for aortocoronary bypass. Forty-five patients (group I) had a single dose of CPC inducing a mean myocardial temperature (MMT) of 32 degrees C/min. Twenty-three patients had multiple doses of CPC and systemic hypothermia to achieve a MMT of 22 degrees C/min. Arterial and coronary sinus sampling 38 minutes after aortic clamp removal permitted calculation of cardiac oxygen extraction, lactate production, CPK and CPK-MB release. Group I patients extracted less oxygen, produced more lactate, and released more CPK and CPK-MB. These indices of cardiac metabolism were found to correlate with anoxic times exceeding 30 minutes, and demonstrated more cardiac damage in group I patients at longer anoxic times. Serial measurements of cardiac output (thermodilution) and left atrial pressure during volume loading permitted construction of myocardial performance curves. Group I patients had a diminished response to volume loading postoperatively. Both the upslope and the highest stroke work attained were lower in group I and inversely related to the anoxic time. All patients made an uneventful recovery, indicating the insensitivity of clinical parameters of myocardial protection. Coronary sinus sampling and hemodynamic monitoring during volume loading permit an objective assessment of myocardial preservation. PMID- 14740706 TI - Resection of the aortic arch using deep hypothermia and temporary circulatory arrest. AB - Resection of the aortic arch has been associated with prohibitive morbidity and mortality because of the technical magnitude of the operation. Six patients, including a 6-year-old boy, recently underwent resection of an aneurysm involving the aortic arch. Deep systemic hypothermia was instituted utilizing cardiopulmonary bypass. Resection of the aneurysm and replacement with a Dacron prosthesis were then performed during a period of total circulatory arrest. During this period the total blood volume of the patient was transferred to the extracorporeal circuit, thus producing a completely dry operating field. Five patients are alive and well although one suffered a transient neurologic deficit from air embolism. The sixth patient died from bleeding. This technique considerably simplifies a difficult operation by producing excellent exposure and eliminating the need for continuous cerebral perfusion. Our experience suggests that resection utilizing deep hypothermia and temporary circulatory arrest is the treatment of choice for patients with aneurysms of the aortic arch. PMID- 14740707 TI - Research challenges assumptions about flame retardant. PMID- 14740708 TI - EU stands firm on chemical regulation overhaul. PMID- 14740709 TI - Buses for the future. PMID- 14740710 TI - Reducing children's risk from lead in soil. PMID- 14740711 TI - Identification of hydroxylated and methoxylated polybrominated diphenyl ethers in Baltic Sea salmon (Salmo salar) blood. AB - Methoxylated and hydroxylated polybrominated diphenyl ethers (MeO-PBDEs and OH PBDEs) have recently been reported to be present in wildlife from Northern Europe. The structures of a majority of these compounds have however been unknown. In the present study, nine OH-PBDEs and six MeO-PBDEs were identified in Baltic Sea salmon (Salmo salar) blood. All OH- and MeO-PBDEs identified were substituted with four or five bromines, and five of these had one chlorine substituent. Fourteen of the OH- and MeO-PBDEs have the methoxy or hydroxy group substituted in the ortho position to the diphenyl ether bond. Identification was done by comparison of relative retention times of authentic reference standards with compounds present in salmon plasma on two gas chromatographic columns of different polarities. The identification was supported by comparisons of full scan mass spectrometric data: electron ionization (EI) and electron capture negative ionization (ECNI). Nine of the 15 OH- and MeO-PBDEs identified have not previously been reported to occur in the environment. The structures of several identified OH- and MeO-PBDEs support natural origin. However, at least one of the OH-PBDEs may be a hydroxylated metabolite of anthropogenic polybrominated diphenyl ether (PBDE). PMID- 14740712 TI - Estimate of oil persisting on the beaches of Prince William Sound 12 years after the Exxon Valdez oil spill. AB - We estimated the amount of oil remaining in Prince William Sound, Alaska, 12 yr after the 1989 Exxon Valdez spill to assess its importance as a long-term reservoir of toxic hydrocarbons. We found oil on 78 of 91 beaches randomly selected according to their oiling history. Surface oiling was recorded for randomly placed quadrats, which were then excavated and examined for subsurface oil. The cumulative area of beach contaminated by surface or subsurface oil was estimated at 11.3 ha. Surface oil varied little with tide height, but subsurface oil was more prevalent at the middle tide heights. The mass of remaining subsurface oil is conservatively estimated at 55 600 kg. Analysis of terpanes indicated that over 90% of the surface oil and all of the subsurface oil was from the Exxon Valdez and that Monterey Formation oil deposited after the 1964 Alaska earthquake accounted for the remaining surface oil. These results indicate that oil from the Exxon Valdez remains by far the largest reservoir of biologically available polycyclic aromatic hydrocarbons on beaches impacted by the spill and that biota dependent on these beaches risk continued exposure. PMID- 14740713 TI - The significance of the North Atlantic Oscillation (NAO) for sea-salt episodes and acidification-related effects in Norwegian rivers. AB - Acidification of Norwegian surface waters, as indicated by elevated concentrations of sulfate and a corresponding reduction in acid neutralizing capacity and pH, is a result of emission and subsequent deposition of sulfur and nitrogen compounds. Episodic sea-salt deposition during severe weather conditions may increase the effects of acidification by mobilizing more toxic aluminum during such episodes. Changes in climatic conditions may increase the frequency and strength of storms along the coast thus interacting with acidification effects on chemistry and biota. We found that the North Atlantic Oscillation (NAO) is linked to sea-salt deposition and sea-salt induced water chemistry effects in five rivers. Particularly, toxic levels of aluminum in all rivers were significantly correlated with higher NAO index values. Further, temporal trends were studied by comparing tendencies for selected statistical indices (i.e. frequency distributions) with time. The selected indices exhibited strong correlations between the NAO index, sea-salt deposition and river data such as chloride, pH and inorganic monomeric aluminum, pointing at the influence of North Atlantic climate variability on water chemistry and water toxicity. The potentially toxic effects of sea-salt deposition in rivers seem to be reduced as the acidification is reduced. This suggests that sea-salt episodes have to increase in strength in order to give the same potential negative biological effects in the future, if acid deposition is further reduced. More extreme winter precipitation events have been predicted in the northwest of Europe as a result of climate change. If this change will be associated with more severe sea-salt episodes is yet unknown. PMID- 14740714 TI - Passive air sampling of PCBs, PBDEs, and organochlorine pesticides across Europe. AB - This study presents concurrently sampled ambient air data for a range of persistent organic pollutants at the continental scale. This was achieved using a passive air sampling system, deploying polyurethane foam disks, which was prepared in one laboratory, sealed to prevent contamination, sent out by courier to volunteers participating in different countries, exposed for 6 weeks, collected, resealed, and returned to the laboratory for analysis. Europe was the study area--a region with a history of extensive POPs usage and emission and with marked national differences in population density, the degree of urbanization and industrial/agricultural development. Samplers were deployed at remote/rural/urban locations in 22 countries and analyzed for PCBs, a range of organochlorine pesticides (HCB, alpha-HCH, gamma-HCH, ppDDT, ppDDE), and PBDEs. Calculated air concentrations were in line with those obtained by conventional active air sampling techniques. The geographical pattern of all compounds reflected suspected regional emission patterns and highlighted localized hotspots. PCB and PBDE levels varied by over 2 orders of magnitude; the highest values were detected in areas of high usage and were linked to urbanized areas. HCB was relatively uniformly distributed, reflecting its persistence and high degree of mixing in air. Higher gamma-HCH, ppDDT, and ppDDE levels generally occurred in South and East Europe. PMID- 14740715 TI - Evaluation of the impact of fuel hydrocarbons and oxygenates on groundwater resources. AB - The environmental behavior of fuel oxygenates (other than methyl tert-butyl ether [MTBE]) is poorly understood because few data have been systematically collected and analyzed. This study evaluated the potential for groundwater resource contamination by fuel hydrocarbons (FHCs) and oxygenates (e.g., tert-butyl alcohol [TBA], tertamyl methyl ether [TAME], diisopropyl ether [DIPE], ethyl tert butyl ether [ETBE], and MTBE) by examining their occurrence, distribution, and spatial extent in groundwater beneath leaking underground fuel tank (LUFT) facilities, focusing on data collected from over 7200 monitoring wells in 868 LUFT sites from the greater Los Angeles, CA, region. Excluding the composite measure total petroleum hydrocarbons as gasoline (TPHG), TBA has the greatestsite maximum (geometric mean) groundwater concentration among the study analytes; therefore, its presence needs to be confirmed at LUFT sites so that specific cleanup strategies can be developed. The alternative ether oxygenates (DIPE, TAME, and ETBE) are less likely to be detected in groundwater beneath LUFT facilities in the area of California studied and when detected are present at lower dissolved concentrations than MTBE, benzene, or TBA. Groundwater plume length was used as an initial indicator of the threat of contamination to drinking water resources. Approximately 500 LUFT sites were randomly selected and analyzed. The results demonstrate MTBE to pose the greatest problem, followed by TBA and benzene. The alternative ether oxygenates were relatively localized and indicated lesser potential for groundwater resource contamination. However, all indications suggest the alternative ether oxygenates would pose groundwater contamination threats similar to MTBE if their scale of usage is expanded. Plume length data suggest that in the absence of a completely new design and construction of the underground storage tank (UST) system, an effective management strategy may involve placing greater emphasis on UST program for ensuring adequate enforcement and compliance with existing UST regulations. PMID- 14740716 TI - Spatial distributions and profiles of atmospheric polycyclic aromatic hydrocarbons in two industrial cities in Japan. AB - The spatial distribution and concentration profiles of 39 vapor and particulate polycyclic aromatic hydrocarbons (PAHs) have been investigated in two Japanese industrial cities (Fuji and Shimizu; a summer and winter season in each). The concentrations of particulate PAHs in winter tended to be higher than those in the summer, but for vapor PAHs, this was not the case. Significant correlations (p < 0.01) were found between most of the PAH concentrations monitored in winter, suggesting the presence of common emission sources. To identify PAH spatial distributions and emission sources in the area, we created contour maps for PAHs monitored; this indicates that the distinctive local distributions correspond to the emission sources. PAH profiles based on benzo[e]pyrene (BeP) concentration, especially for certain relatively heavy molecular weight PAHs, showed differential behaviors among divided areas related to potential regional emission sources such as paper-making plants, power plants, and traffic. We conclude that the origins of atmospheric PAHs in the surveyed areas were dominated by not only traffic but also by stationary emission sources such as paper-making plants and power plants and that local distributions were dependent on the local wind direction. PMID- 14740717 TI - Urinary 1-hydroxypyrene as an indicator for assessing the exposures of booth attendants of a highway toll station to polycyclic aromatic hydrocarbons. AB - In this study, 32 booth attendants (the exposure group) and 21 in pre-job training to become booth attendants (the reference group) were randomly selected from a highway toll station. Personal PAH samplings were conducted on the exposure group on each day during the studied workweek. Pre-shift urinary 1 hydroxylpyrene levels (1-OHP) were measured on the first day of the workweek (BM(pre)) for both the exposure and reference groups, but the post-shift 1-OHP levels were measured on the last day of the workweek (BM(pest)) only for the exposure group. For the exposure group, we found that their mean total PAH exposure level (C(total PAHs)) was 11 400 ng/m3 and that their mean BM(post) was significantly higher than their mean BM(pre) (= 3.02 and 0.910 micromol of 1 OHP/mol of creatinine, respectively). In addition, the mean BM(pre) for the exposure group were higher than that for the reference group (= 0.410 micromol of 1-OHP/mol of creatinine). The above results suggest that vehicle exhaust significantly affects the booth attendants' 1-OHP levels. None of the three personal factors (age, work experience, and smoking habit), except for C(total PAHs), had a significant effect on predicting booth attendants' BM(inc) levels (= BM(post) - BM(pre)) (R2 = 0.57). The above results suggest that urinary 1-OHP could be a suitable biomarker for characterizing workers' PAH exposures. Similarly, we found that none of the three personal characteristics, except for the involved vehicle flow rates and vehicle types, had a significant effect on predicting booth attendants' BM(inc), levels (R2 = 0.60). The above result suggests that the traffic densities of various traffic types could be a suitable surrogate indicator for assessing workers' PAH exposures. Considering that the type of traffic designed for a given type of tollbooth is quite similar all over the world, the results obtained from this study, at least, could be served as a stepping-stone for providing a cheaper and convenient way for assessing traffic PAH exposures in the future. PMID- 14740718 TI - Halonitromethane drinking water disinfection byproducts: chemical characterization and mammalian cell cytotoxicity and genotoxicity. AB - Halonitromethanes are drinking water disinfection byproducts that have recently received a high priority for health effects research from the U.S. Environmental Protection Agency (EPA). Our purpose was to identify and synthesize where necessary the mixed halonitromethanes and to determine the chronic cytotoxicity and the acute genotoxicity of these agents in mammalian cells. The halonitromethanes included bromonitromethane (BNM), dibromonitromethane (DBNM), tribromonitromethane (TBNM), bromochloronitromethane (BCNM), dibromochloronitromethane (DBCNM), bromodichloronitromethane (BDCNM), chloronitromethane (CNM), dichloronitromethane (DCNM), and trichloronitromethane (TCNM). Low- and high-resolution gas chromatography/mass spectrometry (GC/MS) was used to identify the mixed chloro-bromonitromethanes in finished drinking waters, and analytical standards that were not commercially available were synthesized (BDCNM, DBCNM, TBNM, CNM, DCNM, BCNM). The rank order of their chronic cytotoxicity (72 h exposure) to Chinese hamster ovary (CHO) cells was DBNM > DBCNM > BNM > TBNM > BDCNM > BCNM > DCNM > CNM > TCNM. The rank order to induce genomic DNA damage in CHO cells was DBNM > BDCNM > TBNM > TCNM > BNM > DBCNM > BCNM > DCNM > CNM. The brominated nitromethanes were more cytotoxic and genotoxic than their chlorinated analogues. This research demonstrated the integration of the procedures for the analytical chemistry and analytical biology when working with limited amounts of sample. The halonitromethanes are potent mammalian cell cytotoxins and genotoxins and may pose a hazard to the public health and the environment. PMID- 14740719 TI - Chasing quicksilver: modeling the atmospheric lifetime of Hg(0)(g) in the marine boundary layer at various latitudes. AB - The lifetime of elemental mercury in the marine boundary layer(MBL) has been studied using AMCOTS (Atmospheric Mercury Chemistry Over The Sea), a box model of MBL photochemistry including aerosols and detailed mercury chemistry. Recently measured Hg(0)(g) oxidation reactions have been included, and the studies were performed as a function of latitude, time of year, boundary layer liquid water content (LWC) and cloud optical depth. The results show that Hg has the shortest lifetime when air temperatures are low and sunlight and deliquescent aerosol particles are plentiful. Thus the modeled lifetime for clear-sky conditions is actually shorter at mid-latitudes and high latitudes than near the equator, and for a given latitude and time of year, cooler temperatures enhance the rate of Hg oxidation. Under typical summer conditions (for a given latitude) of temperature and cloudiness, the lifetime (tau) of Hg(0)(g) in the MBL is calculated to be around 10 days at all latitudes between the equator and 60 degrees N. This is much shorter than the generally accepted atmospheric residence time for Hg(0)(g) of a year or more. Given the relatively stable background concentrations of Hg(0)(g) which have been measured, continual replenishment of Hg(0)(g) must take place, suggesting a "multihop" mechanism for the distribution of Hg, rather than solely aeolian transport with little or no chemical transformation between source and receptor. Inclusion of an empirical Hg(0)(g) emission factor related to insolation was used to stabilize the Hg(0)(g) concentration in the model, and the emission rates necessarily agree well with estimated emission fluxes for the open ocean. PMID- 14740720 TI - Polycyclic aromatic hydrocarbons in indoor and outdoor environments and factors affecting their concentrations. AB - A highly sensitive analytical method for the simultaneous determination of 39 gaseous and particulate polycyclic aromatic hydrocarbons (PAHs) was used to determine the PAH composition of indoor and outdoor air in Shimizu, Japan. In both indoor and outdoor air, gaseous PAH concentrations were higher in summer than in winter, whereas particulate PAH concentrations were higher in winter than in summer. Correlation analysis indicated that indoor PAH compositions, especially the gaseous PAH composition, differed significantly from outdoor PAH compositions. Gaseous PAH concentrations indoors were significantly affected by insect repellents and heating sources. Particulate PAH concentrations indoors were significantly affected by cigarette smoking, the age and type (wood) of the house, and outdoor PAH concentrations. Inhalation risk associated with carcinogenic PAHs was estimated by using toxic equivalency factors based on the potency of benzo[a]pyrene (BaP). The carcinogenicity of the indoor PAH mixture was dominated by naphthalene followed by BaP and dibenz[a,h]anthracene. PMID- 14740721 TI - Organochlorine compounds in Lake Superior: chiral polychlorinated biphenyls and biotransformation in the aquatic food web. AB - The enantiomeric composition of seven chiral PCB congeners was measured in the Lake Superior aquatic food web sampled in 1998, to determine the extent of enantioselective biotransformation in aquatic biota. All chiral PCB congeners studied (CBs 91, 95, 136, 149, 174, 176, and 183) biomagnified in the Lake Superior aquatic food web, based on biomagnification and food web magnification factors greater than unity. PCB atropisomers were racemic in phytoplankton and zooplankton, suggesting no biotransformation potential toward PCBs for these low trophic level organisms. However, Diporeia and mysids had significantly nonracemic residues for most chiral congeners studied. This observation suggests that these macrozooplankton can stereoselectively metabolize chiral congeners. Alternatively, macrozooplankton obtained nonracemic residues from feeding on organic-rich suspended particles and sediments, which would imply that stereoselective microbial PCB biotransformation may be occurring in Lake Superior sediments at PCB concentrations far lower than that previously associated with such activity. Widely nonracemic PCB residues in forage fish (lake herring, rainbow smelt, and slimy sculpin) and lake trout suggest a combination of both in vivo biotransformation and uptake of nonracemic residues from prey for these species. Minimum biotransformation rates, calculated from enantiomer mass balances between predators and prey, suggest metabolic half-lives on the order of 8 yr for CB 136 in lake trout and 2.6 yr for CB 95 in sculpins. This result suggests that significant biotransformation may occur for metabolizable PCB congeners over the lifespan of these biota. This study highlights the potential of chiral analysis to study biotransformation processes in food webs. PMID- 14740722 TI - Higher brominated diphenyl ethers and hexabromocyclododecane found in eggs of peregrine falcons (Falco peregrinus) breeding in Sweden. AB - Several brominated flame retardants (BFRs) were analyzed in peregrine falcon eggs collected in 1987-1999, including the constituents of the technical polybrominated diphenyl ether (PBDE) products Penta (BDE-47, -99, -100, -153, 154), Octa (BDE-183), and Deca (BDE-209), hexabrominated biphenyl (BB-153), and hexabromocyclododecane (HBCD). The eggs represented females from three different breeding populations, northern Sweden, southwestern Sweden, and a captive breeding population. All BFRs analyzed for were found, including BDE-183 and 209, and concentrations were much higher in wild falcons (geometric mean sigmaPBDE, BB-153, and HBCD for northern/southern populations of 2200/2700, 82/77, and 150/250 ng/g lw, respectively) than in captive falcons (39, 8 ng/g lw, and not detected, respectively). This is the first time, to our knowledge, that BDE-183 and -209 have been quantified in high trophic level wildlife. PMID- 14740723 TI - Source apportionment of sediment PAHs in Lake Calumet, Chicago: application of factor analysis with nonnegative constraints. AB - A factor analysis model with nonnegative constraints (FA) was used to apportion the sources of PAHs found in sediments of Lake Calumet and surrounding wetlands in southeast Chicago. Source profiles and contributions, with uncertainties, are determined with no prior knowledge of sources. The model includes scaling and backscaling of data with average PAH concentrations without sample normalization. This work is a follow-up to a study that used a chemical mass balance (CMB8.2) model to apportion sources to the same data set. Literature source profiles, modified based on gas/particle partitioning of individual PAHs, from eight PAH sources were considered for comparison. FA results for a two-source solution indicate coke oven (45%) and traffic (55%) are the primary PAH sources to Lake Calumet sediments. A six-source FA solution indicates that coke oven (47%) and traffic (45%) related sources are major PAH sources and wood burning-coal residential (2.3%) is a minor PAH source. From the six-source solution, two coke oven profiles are observed, a standard coke oven profile (33%), and a degraded or second coke oven profile (14%) low in phenanthrene and pyrene. Observed traffic related sources include gasoline engine (36%) exhaust and traffic tunnel air (9.3%). This work supports the previous study of Lake Calumet PAHs by CMB model. In addition, FA provides new insights since wood burning and secondary coke oven profiles were not recognized in the CMB model. PMID- 14740724 TI - Bacterial populations associated with the oxidation and reduction of arsenic in an unsaturated soil. AB - Microbial populations responsible for the oxidation and reduction of As were examined in unsaturated (aerobic) soil columns treated with 75 microM arsenite [As(III)] or 250 microM arsenate [As(V)]. Arsenite [As(III)] was rapidly oxidized to As(V) via microbial activity, whereas no apparent reduction of As(V) was observed in the column experiments. Eight aerobic heterotrophic bacteria with varying As redox phenotypes were isolated from the same columns. Three isolates, identified as Agrobacterium tumefaciens-, Pseudomonas fluorescens-, and Variovorax paradoxus-like organisms (based on 16S sequence), were As(III) oxidizers, and all were detected in community DNA fingerprints generated by PCR coupled with denaturing gradient gel electrophoresis. The five other isolates were identified (16S gene sequence) as A. tumefaciens, Flavobacterium sp., Microbacterium sp., and two Arthrobacter sp. -like organisms and were shown to rapidly reduce As(V) under aerobic conditions. Although the two A. tumefaciens like isolates exhibited opposite As redox activity,their 16S rDNA sequences (approximately 1400 bp) were 100% identical, and both were shown to contain putative arsC genes. Our results support the hypothesis that bacteria capable of either oxidizing As(III) or reducing As(V) coexist and are ubiquitous in soil environments, suggesting that the relative abundance and metabolic activity of specific microbial populations plays an important role in the speciation of inorganic As in soil pore waters. PMID- 14740725 TI - Debromination of the flame retardant decabromodiphenyl ether by juvenile carp (Cyprinus carpio) following dietary exposure. AB - The congener 2,2',3,3',4,4',5,5',6,6'-decabromodiphenyl ether (BDE 209) is the primary component in a commonly used flame retardant known as decaBDE. This flame retardant constitutes approximately 80% of the world market demand for polybrominated diphenyl ethers (PBDEs). Because this compound is very hydrophobic (log K(ow) approximately 10), it has been suggested that BDE 209 has very low bioavailability, although debromination to more bioavailable metabolites has also been suggested to occur in fish tissues. In the present study, juvenile carp were exposed to BDE 209 amended food on a daily basis for 60 days, followed by a 40 day depuration period in which the fate of BDE 209 was monitored in whole fish and liver tissues separately. No net accumulation of BDE 209 was observed throughout the experiment despite an exposure concentration of 940 ng/day/fish. However, seven apparent debrominated products of BDE 209 accumulated in whole fish and liver tissues over the exposure period. These debrominated metabolites of BDE 209 were identified as penta- to octaBDEs using both GC/ECNI-MS and GC/HRMS. Using estimation methods for relative retention times of phenyl substitution patterns, we have identified possible structures for the hexa- and heptabromodiphenyl ethers identified in the carp tissues. Although exposure of carp to BDE 209 did not result in the accumulation of BDE 209 in carp tissues, our results indicate evidence of limited BDE 209 bioavailability from food in the form of lower brominated metabolites. PMID- 14740726 TI - Adsorption of natural organic matter to air-water interfaces during transport through unsaturated porous media. AB - To better understand how interactions with the air phase influence the movement of natural organic matter (NOM) through the vadose zone, we measured the transport of soil-humic acid (SHA) through laboratory columns packed with partially saturated sand. Our results demonstrate that sorptive reactions at air water interfaces reduce SHA mobility and that the affinity of SHA for the air phase increases as the porewater pH declines from 8 to 3.9. SHA desorption from air-water interfaces is negligible for conditions of constant pH, but release of bound SHA occurs in response to perturbations in porewater pH. We analyzed the effluent samples collected from our laboratory columns using high-performance size-exclusion chromatography. The results of this analysis demonstrate that the SHA did not fractionate appreciably during transport through the columns, suggesting that the various components of the SHA pool (as distinguished on the basis of molecular weight) express an equal affinity for the air-water interfaces over the range of pH conditions tested. A mathematical model incorporating irreversible, second-order rate laws to simulate adsorption at air-water and solid-water interfaces closely describes the SHA breakthrough data. The mass transfer parameters that govern this model vary in a discernible fashion with changes in porewater pH, and the parameter trends are consistent with published theories for SHA adsorption. PMID- 14740727 TI - Photolytic debromination of decabromodiphenyl ether (BDE 209). AB - Polybrominated diphenyl ethers (PBDE) are commonly used flame retardants. During the past years, concerns have increased due to their occurrence in the environment and humans. In general, the concentrations of lower brominated (tetra penta) diphenyl ethers in biota exceed those of the most heavily used product, decabromodiphenyl ether (DecaBDE). In this study, the photolytic debromination of DecaBDE has been investigated in order to study the formation of lower brominated diphenyl ethers. The time course of photolysis of DecaBDE was studied in toluene, on silica gel, sand, sediment and soil using artificial sunlight and on the natural matrices (sediment, soil, sand) also using natural sunlight. DecaBDE was photolytically labile and formed debromination products in all matrices studied. Nona- to tetraBDEs were formed as well as some PBDFs. The half-lives in toluene and on silica gel were less than 15 min, and half-lives on other matrices ranged between 40 and 200 h. No differences were seen in the debromination pattern of BDE congeners sequentially formed in the different matrices or under different light conditions. However, the debromination rates were strongly dependent on the matrix with longer half-lives on natural matrices than artificial ones. PMID- 14740728 TI - Evolution of toxicity upon wet catalytic oxidation of phenol. AB - This work reports on the evolution of the toxicity of phenol-containing simulated wastewater upon catalytic wet oxidation with a commercial copper-based catalyst (Engelhard Cu-0203T). The results of the study show that this catalyst enhances detoxification, in addition to its effect on the oxidation rate. The EC50 values of the intermediates identified throughout the oxidation route of phenol have been determined and used to predict the evolution of toxicity upon oxidation. The predicted values have been compared with the ones measured directly from the aqueous solution during the oxidation process. To learn about the evolution of toxicity through out the routes of phenol oxidation, experiments have been performed with simulated wastewaters containing separately phenol, catechol, and hydroquinone as original pollutants. The significant increase of toxicity observed during the early stages of phenol oxidation is not directly related to the development of the brown color that derives mainly from catechol oxidation. This increase of toxicity is caused by the formation of hydroquinone and p benzoquinone as intermediates, the former showing the highest toxicity. Furthermore, synergistic effects, giving rise to a significant increase of toxicity, have been observed. These effects derive from the interactions among copper leached from the catalyst and catechol, hydroquinone, and p-benzoquinone and demand that close attention be paid to this potential problem in catalytic wet oxidation. PMID- 14740729 TI - Diversity of contaminant reduction reactions by zerovalent iron: role of the reductate. AB - The reactions of eight model contaminants with nine types of granular Fe(0) were studied in batch experiments using consistent experimental conditions. The model contaminants (herein referred to as "reductates" because they were reduced by the iron metal) included cations (Cu2+), anions (CrO4(2-), NO3(-), and 5,5',7,7' indigotetrasulfonate), and neutral species (2-chloroacetophenone, 2,4,6 trinitrotoluene, carbon tetrachloride, and trichloroethene). The diversity of this range of reductates offers a uniquely broad perspective on the reactivity of Fe(0). Rate constants for disappearance of the reductates vary over as much as four orders of magnitude for particular reductates (due to differences in the nine types of iron) but differences among the reductates were even larger, ranging over almost seven orders of magnitude. Various ways of summarizing the data all suggest that relative reactivities with Fe(0) vary in the order Cu2+, 5,5',7,7'-indigotetrasulfonate > 2-chloroacetophenone, 2,4,6-trinitrotoluene > carbon tetrachloride, CrO4(2-) > trichloroethene > NO3(-). Although the reductate has the largest effect on disappearance kinetics, more subtle differences in reactivity due to the type of Fe(0) suggests that removal of CrO2(2-) and NO3(-) (the inorganic anions) involves adsorption to oxides on the Fe(0), whereas the disappearance kinetics of all other types of reductants is favored by reduction on comparatively oxide-free metal. Correlation analysis of the disappearance rate constants using descriptors of the reductates calculated by molecular modeling (energies of the lowest unoccupied molecular orbitals, LUMO, highest occupied molecular orbitals, HOMO, and HOMO-LUMO gaps) showed that reactivities generally decrease with increasing E(LUMO) and increasing E(GAP) (and, therefore, increasing chemical hardness eta). PMID- 14740730 TI - Sorption of phenanthrene to environmental black carbon in sediment with and without organic matter and native sorbates. AB - Strong sorption to soot- and charcoal-like material (collectively termed black carbon or BC) in soils and sediments is possibly the reason for recent observations of elevated geosorbent-water distribution ratios, slow desorption, limited uptake, and restricted bioremediation. We evaluated the role of environmental BC in the sorption of phenanthrene (PHE) to a polluted lake sediment from a Rhine River sedimentation area. Sorption isotherms were determined over a wide concentration range (0.0005-6 microg/ L) for the original sediment (with organic matter or OM, native sorbates, and BC), sediment from which we had stripped > 90% of the native sorbates (only OM and BC), and sediment combusted at 375 degrees C (only BC). The sorption isotherms of the original and stripped sediments were almost linear (Freundlich coefficient or n(F) > 0.9), whereas the isotherm of the BC remaining after the sediment combustion was highly nonlinear (n(F) = 0.54). At low concentrations (ng/L range), PHE sorption to BC in the combusted sediment was found to exceed the total PHE sorption in the original and stripped sediments. This implies that it may not be possible to use a BC-water sorption coefficient measured in combusted sediment to estimate total sorption to the original sediment. This "intrinsic" BC-water sorption coefficient after combustion was calculated to be 9 times larger than the "environmental" one in the untreated sediment. Competition between the added PHE and the native PAHs and/or OM may explain this difference. It appears that, at low aqueous PHE concentrations (ng/L and below), BC is the most important geosorbent constituent with respect to sorption. At higher concentrations (microg/L), BC sorption sites become saturated and BC sorption is overwhelmed by sorption to the other OM constituents. Because sorption is a central process affecting contaminant behavior and ecotoxicity, understanding this process can strongly contribute to risk assessment and fate modeling. PMID- 14740731 TI - Understanding the effects of soil characteristics on phytotoxicity and bioavailability of nickel using speciation models. AB - Acidity (pH) has been realized to be the most important soil characteristic that modulates bioavailability of heavy metals by affecting both the chemical speciation of metals in soil and the metal binding to the active sites on biota. In this work, we show that besides soil pH, metal bioavailability also depends to a certain extent on the type of soil. A better understanding of the role of soil type in regulating metal availability can be achieved with the analysis of soil composition and with calculations using chemical speciation models. Results of pot experiments, in which three different soils were spiked with nickel, show that the EC50 of total nickel in decreasing the biomass production of oats varies widely (0.7-22.5 mmol kg(-1) soil, more than 30 times). pH (4.7-7.0) is the most important factor, explaining up to a factor of 14 difference of nickel bioavailability in the soils. The remaining variation is caused by other differences in soil composition (soil type). The bioavailability and toxicity of nickel in the organic matter-rich soil studied is less than half of that in the sandy and clay soil studied at a similar pH. The chemical calculations using a multi-surface speciation model show that soil organic matter binds Ni much stronger than clay silicates and iron (hydr)oxides within the acidic pH range, which supports the experimental findings. In all three soils, the EC50 of Ni expressed in terms of Ni in 0.01 M CaCl2 soil extraction is rather stable (24-58 microM), suggesting the possibility to use this extraction as an estimation of metal availability in soil. PMID- 14740732 TI - 2,4-D sorption in iron oxide-rich soils: role of soil phosphate and exchangeable Al. AB - This study examined herbicide retention in iron oxide-rich variable charge soils (Ultisols) under no cultivation (forest), agriculture (farm), and turf maintenance (golf course) to explore the following hypothesis: inorganic phosphate accumulation from soil fertilization and liming to decrease exchangeable aluminum (Al) content will influence carboxylic acid herbicide sorption onto soils and leaching into groundwater. A suite of soil properties, including mineralogy (particularly soil iron and aluminum oxide content), exchangeable Al content, and soil phosphate content, influenced sorption of the anionic, 2,4-D. In general, 2,4-D sorption was lower in the presence of phosphate, possibly due to competition between phosphate and 2,4-D for surface sites or increase in surface negative charge resulting from phosphate sorption. Additionally, 2,4-D sorption was greater in the presence of exchangeable Al. It appears that 2,4-D may form surface complexes with or be electrostatically attracted to exchangeable aluminum in the soil. Our results suggest that carboxylic acid herbicides may be more easily leached in intensively managed Ultisols subject to continued phosphate fertilization and liming. PMID- 14740733 TI - Influence of dissolved sodium and cesium on uranyl oxide hydrate solubility. AB - The solubility of uranium-containing minerals can control the mobility of uranium in contaminated soil and groundwater. The identity and solubility of these minerals are strongly influenced by solution composition. The influence of dissolved sodium and cesium on the solubility of uranyl oxide hydrates has been investigated in a series of batch experiments conducted with synthetic metaschoepite ((UO2)8O2(OH)12 x 10H2O). During reaction of metaschoepite in NaNO3, CsNO3, and NaF solutions, an initial increase in the dissolved uranium concentration was followed by a decrease as uranium was incorporated into a secondary solid phase. Given sufficient reaction time, metaschoepite was completely transformed to a clarkeite-like sodium uranyl oxide hydrate or a cesium uranyl oxide hydrate that has not previously been described. These secondary solid phases exhibited X-ray diffraction patterns and Raman spectra that were distinct from those of the original metaschoepite. Dissolved uranium concentrations in equilibrium with the sodium and cesium uranyl oxide hydrates can be more than 2 orders of magnitude lower than those in equilibrium with metaschoepite. Initial changes in metaschoepite solubility may also result from particle growth induced by sodium and cesium incorporation into the solid phase. PMID- 14740734 TI - Adsorption of the herbicide simazine by montmorillonite modified with natural organic cations. AB - Three organic cations with a natural origin (L-carnitine, L-cystine dimethyl ester, and thiamine) were introduced at different loadings in the interlayer of a low-charge montmorillonite, and the performance of the modified clays as adsorbents of the herbicide simazine was investigated using batch adsorption desorption experiments. The organic cations were selected on the basis of their natural origin and the presence of diverse functional groups in their structures, which was expected to influence simazine adsorption. Elemental analysis and spectroscopy results demonstrated the presence of the organic cations in the modified montmorillonites and their entrance in the clay mineral interlayers. Batch adsorption results showed that modification with thiamine (K(f) = 96-138), cystine dimethyl ester (K(f) = 400-753), and especially carnitine (K(f) > 10 000) enhanced the adsorption of simazine by montmorillonite (K(f) = 28-47). It appeared that the specific interlayer microenvironment provided by the functional groups of each organic cation was an important factor controlling the adsorption efficiency of the modified clays. For carnitine and cystine dimethyl ester, the increase in simazine adsorption was considerably greater than that observed after montmorillonite modification with "classical" alkylammonium cations, such as phenyltrimethylammonium or hexadecyltrimethylammonium. This illustrated how modification of smectitic clay minerals with natural organic cations containing appropriate functional groups can be a useful strategy to improve the performance of organoclays for the removal of specific organic pollutants from the environment. PMID- 14740735 TI - Inhibition of biological reductive dissolution of hematite by ferrous iron. AB - Bacterial dissimilatory iron reduction is self-inhibited by the production of ferrous [Fe(II)] iron resulting in diminished iron reduction as Fe(II) accumulates. Experiments were conducted to investigate the mechanisms of Fe(II) inhibition employing the dissimilatory metal-reducing bacterium Shewanella putrefaciens strain CN32 under nongrowth conditions in a system designed to minimize precipitation of ferrous iron minerals. After an initial period (ca. 1 day) of relatively rapid iron reduction, hematite reduction rates were controlled by mass transfer of Fe(II). Experiments in which hematite was equilibrated with Mn(II) prior to inoculation indicated that the observed inhibition was not due to Fe(II) sorption. At longer times, soluble Fe(II) accumulated such that the reaction was slowed due to a decreased thermodynamic driving force. The thermodynamic evaluation also supported the prior conclusion that hydrated hematite surface sites may yield substantially more energy during bioreduction than "bulk" hematite. For well-mixed conditions, the rates of hematite reduction were directly proportional to the biologically available reaction potential. PMID- 14740736 TI - Dissipation of fragrance materials in sludge-amended soils. AB - A possible removal mechanism for fragrance materials (FMs) in wastewater is adsorption to sludge, and sludge application to land may be a route through which FMs are released to the soil environment. However, little is known about the concentrations and fate of FMs in soil receiving sludge application. This study was conducted to better understand the dissipation of FMs in sludge-amended soils. We first determined the spiking and extraction efficiencies for 22 FMs in soil and leachate samples. Nine FMs were detected in digested sludges from two wastewater treatment plants in Delaware using these methods. We conducted a 1 year die-away experiment which involved four different soils amended with sludge, with and without spiking of the 22 FMs. The initial dissipation of FMs in all spiked trays was rapid, and only seven FMs remained at concentrations above the quantification limits after 3 months: AHTN, HHCB, musk ketone, musk xylene, acetyl cedrene, OTNE, and DPMI. After 1 year, the only FMs remaining in all spiked trays were musk ketone and AHTN. DPMI was the only FM that leached significantly from the spiked trays, and no FMs were detected in leachate from any unspiked tray. While soil organic matter content affected the dissipation rate in general, different mechanisms (volatilization, transformation, leaching) appeared to be important for different FMs. PMID- 14740737 TI - Analysis of ambient particle size distributions using Unmix and positive matrix factorization. AB - Hourly averaged particle size distributions measured at a centrally located urban site in Seattle were analyzed through the application of bilinear positive matrix factorization (PMF) and Unmix to study underlying size distributions and their daily patterns. A total of 1051 samples each with 16 size intervals from 20 to 400 nm were obtained from a differential mobility particle sizer operating between December 2000 and February 2001. Both PMF and Unmix identify four similar underlying factors in the size distributions. Factor 1 is an accumulation mode particle size spectrum that shows a regular nocturnal pattern, and factor 2 is a larger particle distribution. Factor 3 is assigned as a traffic-related particle distribution, based on its correlations with accompanying gas-phase measurements, and has a regular weekday-high rush-hour pattern. Factor 4 is a traffic-related particle size distribution that has a regular rush-hour pattern on weekdays as well as weekends. Conditional probability functions (CPF) were computed using wind profiles and factor contributions. The results of CPF analysis suggest that these factors are correlated with surrounding particle sources of wood burning, secondary aerosol, diesel emissions, and motor vehicle emissions. PMID- 14740738 TI - Deposition and reentrainment of Brownian particles in porous media under unfavorable chemical conditions: some concepts and applications. AB - The deposition and reentrainment of particles in porous media have been examined theoretically and experimentally. A Brownian Dynamics/Monte Carlo (MC/BD) model has been developed that simulates the movement of Brownian particles near a collector under "unfavorable" chemical conditions and allows deposition in primary and secondary minima. A simple Maxwell approach has been used to estimate particle attachment efficiency by assuming deposition in the secondary minimum and calculating the probability of reentrainment. The MC/BD simulations and the Maxwell calculations support an alternative view of the deposition and reentrainment of Brownian particles under unfavorable chemical conditions. These calculations indicate that deposition into and subsequent release from secondary minima can explain reported discrepancies between classic model predictions that assume irreversible deposition in a primary well and experimentally determined deposition efficiencies that are orders of magnitude larger than Interaction Force Boundary Layer (IFBL) predictions. The commonly used IFBL model, for example, is based on the notion of transport over an energy barrier into the primary well and does not address contributions of secondary minimum deposition. A simple Maxwell model based on deposition into and reentrainment from secondary minima is much more accurate in predicting deposition rates for column experiments at low ionic strengths. It also greatly reduces the substantial particle size effects inherent in IFBL models, wherein particle attachment rates are predicted to decrease significantly with increasing particle size. This view is consistent with recent work by others addressing the composition and structure of the first few nanometers at solid-water interfaces including research on modeling water at solid-liquid interfaces, surface speciation, interfacial force measurements, and the rheological properties of concentrated suspensions. It follows that deposition under these conditions will depend on the depth of the secondary minimum and that some transition between secondary and primary depositions should occur when the height of the energy barrier is on the order of several kT. When deposition in secondary minima predominates, observed deposition should increase with increasing ionic strength, particle size, and Hamaker constant. Since an equilibrium can develop between bound and bulk particles, the collision efficiency [alpha] can no longer be considered a constant for a given physical and chemical system. Rather, in many cases it can decrease over time until it eventually reaches zero as equilibrium is established. PMID- 14740739 TI - Intercomparison of volatile organic carbon measurement techniques and data at La Porte during the TexAQS2000 Air Quality Study. AB - The Texas Air Quality Study 2000 (TexAQS2000) investigated the photochemical production of ozone and the chemistry of related precursors and reaction products in the vicinity of Houston, TX. The colocation of four instruments for the measurement of volatile organic carbon compounds (VOCs) allowed a unique opportunity for the intercomparison of the different in-situ measuring techniques. The instruments included three gas chromatographs, each with a different type of detector, and a Proton-Transfer-Reaction Mass Spectrometer (PTR MS) with each system designed to measure a different suite of VOCs. Correlation plots and correlation statistics are presented for species measured by more than one of these instruments. The GC instruments were all in agreement to within 10 20% (slope) with coefficients of variation (r2) of > or = 0.85. The PTR-MS agreement with other instruments was more dependent on species with some very good agreements (r2 values of approximately 0.95 for some aromatics), but isoprene, acetaldehyde and propene were substantially less highly correlated (0.55 < r2 < 0.80). At least part of these differences were undoubtedly due to the timing of sample acquisition in an environment in which VOC levels changed very rapidly on both quantitative and temporal scales. PMID- 14740740 TI - Identification of the non-pertechnetate species in Hanford waste tanks, Tc(I)- carbonyl complexes. AB - Immobilization of the high-level nuclear waste stored at the Hanford Reservation has been complicated by the presence of soluble, lower-valent technetium species. Previous work by Schroeder and Blanchard has shown that these species cannot be removed by ion-exchange and are difficult to oxidize. The Tc K-edge XANES spectra of the species in Tanks SY-101 and SY-103 were reported by Blanchard, but they could not be assigned to any known technetium complex. We report that the XANES spectra are most likely those of Tc(I)-carbonyl species, especially fac Tc(CO)3(gluconate)(2-). This is further supported by EXAFS and 99Tc-NMR studies in nonradioactive simulants of these tank wastes. PMID- 14740741 TI - Field screening of waterborne petroleum hydrocarbons by thickness shear-mode resonator measurements. AB - An inexpensive, field-portable sensor for direct, aggregate determination of aqueous petroleum hydrocarbons (PH) down to sub-ppm levels was developed. The basis of this sensor was an unusual, highly nongravimetric frequency response of 10 MHz (series fundamental) AT-cut quartz crystals when coated with rubbery silicone films. The response depended linearly and reliably on the total concentration of dissolved hydrocarbons over a range of 0.01-100 mg x L(-1) or up to aqueous solubility limits. Calibration sensitivities were measured individually for laboratory-prepared solutions of BTEX (benzene, toluene, ethylbenzene, and xylene isomers) and C6-C8 aliphatic components. Each component demonstrated a method detection limit (MDL) in the low-to sub-ppm range (benzene 10 mg x L(-1), n-hexane 0.54 mg x L(-1)) for light coatings of a commercially available poly-(dimethylsiloxane) gum (OV-1, > 10(6) g x mol(-1)) and lower MDLs for heavier coatings. Pairwise responses for the aliphatic and benzenoid standards were additive, indicating that aggregate determinations of mixtures (especially light fuels) were possible. Natural matrix interferences caused by sample turbidity and ionic strength were overcome by simple preparative methods. Fuel-spiked natural waters were determined with respect to standards and verified by gas chromatography. A 0.19 mg x L(-1) MDL for gasoline was obtained for heavy OV-1 films. Field determinations of groundwater surrounding a leaking underground fuel tank demonstrated that the sensor and method were useful for on-site PH screening. Large differences between the equilibration times of aliphatic and benzenoid components also indicated one avenue for BTEX speciation with the device. PMID- 14740742 TI - Methods for estimating adsorbed uranium(VI) and distribution coefficients of contaminated sediments. AB - Assessing the quantity of U(VI) that participates in sorption/desorption processes in a contaminated aquifer is an important task when investigating U migration behavior. U-contaminated aquifer sediments were obtained from 16 different locations at a former U mill tailings site at Naturita, CO (U.S.A.) and were extracted with an artificial groundwater, a high pH sodium bicarbonate solution, hydroxylamine hydrochloride solution, and concentrated nitric acid. With an isotopic exchange method, both a KD value for the specific experimental conditions as well as the total exchangeable mass of U(VI) was determined. Except for one sample, KD values determined by isotopic exchange with U-contaminated sediments that were in equilibrium with atmospheric CO2 agreed within a factor of 2 with KD values predicted from a nonelectrostatic surface complexation model (NEM) developed from U(VI) adsorption experiments with uncontaminated sediments. The labile fraction of U(VI) and U extracted by the bicarbonate solution were highly correlated (r2 = 0.997), with a slope of 0.96 +/- 0.01. The proximity of the slope to one suggests that both methods likely access the same reservoir of U(VI) associated with the sediments. The results indicate that the bicarbonate extraction method is useful for estimating the mass of labile U(VI) in sediments that do not contain U(IV). In-situ KD values calculated from the measured labile U(VI) and the dissolved U(VI) in the Naturita alluvial aquifer agreed within a factor of 3 with in-situ KD values predicted with the NEM and groundwater chemistry at each well. PMID- 14740743 TI - Development of a method to assess cigarette smoke intake. AB - Tar and nicotine deliveries of cigarettes measured using current standardized smoking machine protocols provide poor estimates of smoke exposure. The characteristics of human smoking behavior vary considerably and differ from the rigid parameters used with current standardized smoking machine protocols. Current alternatives, including measurement of biomarkers, are invasive, time dependent, and can be too expensive to be used as mechanisms for carrying out large-scale investigations required to help determine the influence of cigarette design on smoking behaviors. To obtain more reasonable estimates of mainstream smoke exposure, we developed a method to quantitatively measure solanesol, a naturally occurring component in tobacco that is deposited during smoking in the cigarette filter butt. Quantification of solanesol extracted from the filters using liquid chromatography and tandem mass spectrometry is efficient, rapid, and extremely reliable. We found that the amount of solanesol deposited in a cigarette filter is related to the mainstream smoke deliveries of tar and nicotine under a variety of smoking conditions. In addition, the amount of solanesol trapped in the filter remains stable at least 4 weeks after smoking. Measuring solanesol in cigarette filters as an exposure marker provides a noninvasive means to obtain reasonable estimates of mainstream tar and nicotine smoke deliveries under a wide variety of smoking conditions. PMID- 14740744 TI - Sampling artifacts of acidity and ionic species in PM2.5. AB - Although sampling artifacts of acidity, ammonium, nitrate, and chloride in airborne particulate pollutants can be reduced by the use of denuders to absorb interfering gases, artifacts due to interparticle interactions still remain. In this study, the contribution of individual artifact reactions to particle evaporation and the effects of aerosol composition on the extents of sampling artifacts in PM2.5 were investigated. Samples were collected using a Harvard honeycomb denuder/filter-pack system at an urban site and a rural site in Hong Kong. The results show that the formation of artifacts can be categorized into two regimes: ammonium rich (AR) samples with a molar ratio [NH4+]/ [SO4(2-)] greater than 1.5 and ammonium poor (AP) samples with a molar ratio [NH4+]/[SO4(2 )] less than or equal to 1.5. The urban samples were all AR samples, and they were characterized by high nitrate and low in situ free acid concentrations. In contrast, the rural samples were all AP samples and they were characterized by low nitrate and high in situ free acid concentrations. We have developed a methodology to estimate the contribution of each artifact reaction to the sampling loss of nitrate, chloride, ammonium, and acidity. In the AR samples, the evaporation of HNO3 and HCl and concomitant evaporation of NH3 were the principal reactions in determining the extent of the sampling loss of nitrate and chloride. In the AP samples, the evaporation of HNO3 and HCl alone was the principal reaction instead, especially at high sampling loss. The in situ free acid concentration, a function of aerosol composition and ambient conditions, is a more useful parameter than strong acidity in understanding the sampling loss of acidity, nitrate, and chloride from the collected particles. PMID- 14740745 TI - Synergistic effect of copper ion on the reductive dechlorination of carbon tetrachloride by surface-bound Fe(II) associated with goethite. AB - The dechlorination of carbon tetrachloride (CT) by Fe(II) associated with goethite in the presence of transition metal ions was investigated. X-ray photoelectron spectroscopy (XPS) and X-ray powder diffraction (XRPD) were used to characterize the chemical states and crystal phases of transition metals on solid phases, respectively. CT was dechlorinated to chloroform (CF) by 3 mM Fe(II) in 10 mM goethite (25.6 m2 L(-1)) suspensions. The dechlorination followed pseudo first-order kinetics, and a rate constant (k(obs)) of 0.036 h(-1) was observed. Transition metal ions have different effects on CT dechlorination. The addition of Ni(II), Co(II), and Zn(II) lowered the k(obs) for CT dechlorination, whereas the amendment of 0.5 mM Cu(II) into the Fe(II)-Fe(III) system significantly enhanced the efficiency and the rate of CT dechlorination. The k(obs) for CT dechlorination with 0.5 mM Cu(II) was 1.175 h(-1), which was 33 times greater than that without Cu(II). Also, the dechlorination of CT by surface-bound iron species is pH-dependent, and the rate constants increased from 0.008 h(-1) at pH 4.0 to 1.175 h(-1) at pH 7.0. When the solution contained Cu(II) and Fe(II) without goethite, a reddish-yellow precipitate was formed, and the concentration of Fe(ll) decreased with the increase in Cu(II) concentration. XPS and XRPD analyses suggested the possible presence of Cu2O and ferrihydrite in the precipitate. Small amounts of aqueous Cu(I) were also detected, reflecting the fact that Cu(II) was reduced to Cu(I) by Fe(II). A linear relationship between k(obs) for CT dechlorination and the concentration of Cu(II) was observed when the amended Cu(II) concentration was lower than 0.5 mM. Moreover, the k(obs) for CT dechlorination was dependent on the Fe(II) concentration in the 0.5 mM Cu(II) amended goethite system and followed a Langmuir-Hinshelwood relationship. These results clearly indicate that Fe(II) serves as the bulk reductant to reduce both CT and Cu(II). The resulting Cull) can further act as a catalyst to enhance the dechlorination rate of chlorinated hydrocarbons in iron-reducing environments. PMID- 14740746 TI - Novel bentonite clay-based Fe-nanocomposite as a heterogeneous catalyst for photo Fenton discoloration and mineralization of Orange II. AB - A novel bentonite clay-based Fe-nanocomposite (Fe-B) was successfully developed as a heterogeneous catalyst for photo-Fenton discoloration and mineralization of an azo-dye Orange II. X-ray diffraction (XRD) analysis clearly reveals that the Fe-B nanocomposite catalyst mainly consists of Fe2O3 (hematite) and SiO2 (quartz) crystallites, and the Fe concentration of the Fe-B catalyst determined by X reflective fluorescence (XRF) is 31.8 wt %. The catalytic activity of the Fe-B was evaluated in the discoloration and mineralization of Orange II in the presence of H2O2 and UVC light (254 nm). It was found that the optimal Fe-B catalyst dosage is around 1.0 g/L, and the efficiency of discoloration and mineralization of Orange II increases as initial Orange II concentration decreases or reaction temperature increases. In addition, at optimal conditions (10 mM H2O2, 1.0 g of Fe-B/L, 1 x 8W UVC, and pH = 3.0), complete discoloration and mineralization of 0.2 mM Orange II can be achieved in less than 60 and 120 min, respectively. The result strongly indicates that the Fe-B nanocomposite catalyst exhibits a high catalytic activity not only in the photo-Fenton discoloration of Orange II but also in the mineralization of Orange II. The reaction kinetics analysis illustrates that the photo-Fenton discoloration of Orange II in the first 15 min obeys the pseudo-first-order kinetics. The reaction activation energy calculated was 9.94 kJ/mol, indicating that the photo-Fenton discoloration of Orange II is not very sensitive to reaction temperature. PMID- 14740747 TI - Enhancing activated carbon adsorption of 2-methylisoborneol: methane and steam treatments. AB - This research investigated methods for tailoring a commercial, lignite-based granular activated carbon (GAC) to enhance its adsorption of 2-methylisoborneol (MIB) from natural water. Tailoring efforts focused on heat treatments in gas environments comprising steam and/or methane, since these gases can alter GAC pore structure and surface chemistry. Heat treatments that combined methane and steam enhanced MIB adsorption considerably, causing a 4-fold improvement (over untreated GAC) in fixed-bed adsorption performance relative to initial MIB breakthrough. These favorable effects, observed in rapid small-scale column tests, occurred following simultaneous and separate (sequential) applications of methane and steam. Moderately low temperature steam treatments also improved MIB uptake in fixed-bed adsorption tests but to a lesser extent (approximately 1.5 fold). In contrast, methane treatments alone, at various temperatures, led to significant carbon deposition within the GAC pore structure. As a result, total pore volume was reduced and MIB adsorption performance declined. PMID- 14740748 TI - Improvement of catalyst durability by deposition of Rh on TiO2 in photooxidation of aromatic compounds. AB - Gas-solid heterogeneous photocatalytic oxidation of aromatic compounds in air was carried out at room temperature with a fixed bed flow reactor. The deposition of Rh on TiO2 catalyst improved the catalyst durability in benzene photooxidation. The Rh deposition reduced the amounts of carbonaceous materials on the catalyst surface that were the cause of catalyst deactivation. The highest reaction rate was obtained at the Rh loading of 0.5-1.0 wt %. The Rh/TiO2 catalyst was gradually deactivated in prolonged benzene photooxidation, due to the increasing amount of carbonaceous materials on the catalyst surface. XPS studies showed that the gradual deactivation was related to the changes in the oxidation state of the surface Rh metals. The catalysts were regenerated by the treatment of hydrogen reduction after the photoirradiation in humidified air, indicating that Rh(0) was essential for the improvement of the catalyst durability. The catalyst was also regenerated by the heat treatment in N2 flow instead of the hydrogen reduction. Rh deposition was also effective for the improvement of catalyst durability in toluene photooxidation. PMID- 14740749 TI - Phytophotolysis of hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) in leaves of reed canary grass. AB - Hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) was degraded in reed canary grass leaves exposed to simulated sunlight to primary products nitrous oxide and 4 nitro-2,4-diazabutanal. This is the first time that 4-nitro-2,4-diazabutanal, a potentially toxic degradate, has been measured in plant tissues following phytotransformation of RDX. These compounds, along with nitrite and formaldehyde, were also detected in aqueous RDX systems exposed to the same simulated sunlight. Results showed that the initial products of RDX photodegradation in translucent plant tissues were similar to products formed from aqueous photolysis of RDX. Combustion analysis of leaves following 14C-RDX uptake and subsequent light exposure revealed the presence of tissue-bound material that could not be extracted with acetonitrile. No detectable formaldehyde was emitted from the leaves. The detection of similar RDX degradation products in both aqueous and plant-based systems suggests that RDX may be initially transformed by similar mechanisms in both systems. Direct photolysis of RDX via ultraviolet irradiation passing into the leaves is hypothesized to be responsible for the observed transformations. In addition, membrane-bound "trap chlorophyll" in the chloroplasts may shuttle electrons to RDX as an indirect photolysis transformation mechanism. Results from this study indicate that reed canary grass facilitates photochemical degradation of RDX, and this mechanism should be considered along with more established phytoremediation processes when assessing the fate of contaminants in plant tissues. Plant-mediated phototransformation of xenobiotic compounds is a process that may be termed "phytophotolysis". PMID- 14740750 TI - Photoproduction of hydroxyl radicals in aqueous solution with algae under high pressure mercury lamp. AB - Photoproduction of hydroxyl radicals (*OH) could be induced in aqueous solution with algae (Nitzschia hantzschiana, etc.) and (or not) Fe3+ under high-pressure mercury lamp with an exposure time of 4 h. *OH was determined by HPLC using benzene as a probe. The photoproduction of *OH increased with increasing algae concentration. Fe3+ could enhance the photoproduction of *OH in aqueous solution with algae. The results showed that the photoproduction of *OH in algal solution with Fe3+ was greater than that in algal solution without Fe3+. The light intensity and pH affected the photoproduction of *OH in aqueous solution with algae with/without Fe3+. The photoproduction of *OH in aqueous solution with algae and Fe3+ under 250 W was greater than that under 125 W HPML. The photoproduction of *OH in algal solution (pH ranged from 4.0 to 7.0) with (or not) Fe3+ at pH 4 was the greatest. PMID- 14740751 TI - Bioaccumulation of chromium from tannery wastewater: an approach for chrome recovery and reuse. AB - The presence of chromium in the effluent is a major concern for the tanning industry. Currently, chemical precipitation methods are practiced for the removal of chromium from the effluent, but that leads to the formation of chrome-bearing solid wastes. The other membrane separation and ion exchange methods available are unfeasible due to their cost. In this study, the removal of chromium from tannery effluent has been carried out using abundantly available brown seaweed Sargassum wightii. Simulated chrome tanning solution was used for the standardization of experimental trials. Various factors influencing the uptake of chromium, viz., quantity of seaweed, concentrations of chromium, pH of the chrome bearing wastewater, and duration of treatment, have been studied. Chemical modification of the seaweed through pretreatment with sulfuric acid, magnesium chloride, and calcium chloride showed improved uptake of chromium. Langmuir and Freundlich isotherms have been fitted for various quantities of seaweed. The dynamic method of treatment of protonated seaweed with simulated chrome tanning solution at a pH of 3.5-3.8 for a duration of 6 h gave the maximum uptake of about 83%. A similar uptake has been established for commercial chrome tanning wastewater containing the same concentration of chromium. The Sargassum species exhibited a maximum uptake of 35 mg of chromium per gram of seaweed. Fourier transform infrared spectroscopy, energy-dispersive X-ray analysis, and flame photometry studies have been carried out to understand the mechanistic pathway for the removal of chromium. The potential reuse of chromium-containing seaweed for the preparation of basic chromium sulfate (tanning agent) has been demonstrated. PMID- 14740752 TI - Arsenic removal with iron(II) and iron(III) in waters with high silicate and phosphate concentrations. AB - Arsenic removal by passive treatment, in which naturally present Fe(II) is oxidized by aeration and the forming iron(III) (hydr)oxides precipitate with adsorbed arsenic, is the simplest conceivable water treatment option. However, competing anions and low iron concentrations often require additional iron. Application of Fe(II) instead of the usually applied Fe(III) is shown to be advantageous, as oxidation of Fe(II) by dissolved oxygen causes partial oxidation of As(III) and iron(III) (hydr)oxides formed from Fe(II) have higher sorption capacities. In simulated groundwater (8.2 mM HCO3(-), 2.5 mM Ca2+, 1.6 mM Mg2+, 30 mg/L Si, 3 mg/L P, 500 ppb As(III), or As(V), pH 7.0 +/- 0.1), addition of Fe(II) clearly leads to better As removal than Fe(III). Multiple additions of Fe(II) further improved the removal of As(II). A competitive coprecipitation model that considers As(III) oxidation explains the observed results and allows the estimation of arsenic removal under different conditions. Lowering 500 microg/L As(III) to below 50 microg/L As(tot) in filtered water required > 80 mg/L Fe(III), 50-55 mg/L Fe(II) in one single addition, and 20-25 mg/L in multiple additions. With As(V), 10-12 mg/L Fe(II) and 15-18 mg/L Fe(III) was required. In the absence of Si and P, removal efficiencies for Fe(II) and Fe(III) were similar: 30-40 mg/L was required for As(II), and 2.0-2.5 mg/L was required for As(V). In a field study with 22 tubewells in Bangladesh, passive treatment efficiently removed phosphate, but iron contents were generally too low for efficient arsenic removal. PMID- 14740753 TI - Influence of surface properties on the mechanism of H2S removal by alkaline activated carbons. AB - Alkaline activated carbons are widely used as adsorbents of hydrogen sulfide (H2S), one of the major odorous compounds arising from sewage treatment facilities. Although a number of studies have explored the effects of various parameters, mechanisms of H2S adsorption by alkaline carbons are not yet fully understood. The major difficulty seems to lie in the fact that little is known with certainty about the predominant reactions occurring on the carbon surface. In this study, the surface properties of alkaline activated carbons were systematically investigated to further exploit and better understand the mechanisms of H2S adsorption by alkaline activated carbons. Two commercially available alkaline activated carbons and their representative exhausted samples (8 samples collected at different height of the column after H2S breakthrough tests) were studied. The 8 portions of the exhausted carbon were used to represent the H2S/carbon reaction process. The surface properties of both the original and the exhausted carbons were characterized using the sorption of nitrogen (BET test), surface pH, Boehm titration, thermal and FTIR analysis. Porosity and surface area provide detailed information about the pore structure of the exhausted carbons with respect to the reaction extent facilitating the understanding of potential pore blockages. Results of Boehm titration and FTIR both demonstrate the significant effects of surface functional groups, and identification of oxidation products confirmed the different mechanisms involved with the two carbons. From the DTG curves of thermal analysis, two well-defined peaks representing two products of surface reactions (i.e., sulfur and sulfuric acid) were observed from the 8 exhausted portions with gradually changing patterns coinciding with the extent of the reaction. Surface pH values of the exhausted carbons show a clear trend of pH drop along the reaction extent, while pH around 2 was observed for the bottom of the bed indicating sulfuric acid as the predominant products. Although both carbons are coal-based and of KOH impregnated type, performances of different carbons differ significantly. A correlation is well established to link the reaction extent with various surface properties. In summary, not only the homogeneous alkali impregnation and physical porosity but also the carbon surface chemistry are significant factors influencing the performances of alkaline activated carbons as H2S adsorbents. PMID- 14740754 TI - Collagen fiber immobilized Myrica rubra tannin and its adsorption to UO2(2+). AB - Tannins, which are rich in ortho-hydroxyl groups, have a high affinity for UO2(2+). In this paper, Myrica rubra tannin was immobilized on collagen fiber by an aldehydic cross-linking reaction to prepare a novel adsorbent for uranium (UO2(2+)) recovery from wastewater. The adsorption equilibrium, the adsorption kinetics, and the effects of temperature and pH on the adsorption equilibrium were investigated in detail. It was found that the Myrica rubra tannin immobilized on collagen fiber exhibits an excellent adsorption capacity for UO2(2+). The adsorption capacity at 293 K and pH 5.0 was as high as 1.19 mmol UO2(2+)/g (283.3 mgU/g) when the initial concentration of UO2(2+) in solution was 7.5 mmol/L. The adsorption isotherms could be described by the Freundlich equation, and the increase of temperature promoted the adsorption to UO2(2+) . The adsorption kinetics data were fitted very well by the pseudosecond-order rate model, and the equilibrium adsorption capacity calculated by the pseudo-second order rate model was almost the same as that determined by the actual measurement with the error < or = 4%. The pH has a significant effect on the adsorption process. According to our experiments, the suitable pH scope should be 5-8. PMID- 14740755 TI - Photocatalysis by titanium dioxide and polyoxometalate/TiO2 cocatalysts. Intermediates and mechanistic study. AB - A representative polyoxometalate, alpha-12-tungstophosphatic acid (PW12(3-), POM), is loaded on the surface of TiO2 particles used as a cocatalyst to gain further insights into the underlying reaction mechanism and to estimate the feasibility of using the new POM/TiO2 cocatalyst in the photocatalytic degradation of 2,4-dichlorophenol (DCP) in aqueous media. Loading the PW12(3-) species on the surface of TiO2 enhances charge separation in the UV-illuminated TiO2, thereby accelerating the hydroxylation of the initial DCP substrate but not the mineralization of DCP, which is somewhat suppressed in the presence of the polyoxometalate. An increase in the load of POM increases the concentration of aromatic intermediates, and more toxic intermediates, such as 2,6-dichlorodibenzo p-dioxin, 2,4,6-trichlorophenol, are detected in the PW12(3-)/TiO2 system. By contrast, cleavage of the whole conjugated structure of DCP predominates in TiO2 only dispersions. Strong ESR signals for the superoxide radical anionic species, O2*- (HO2* radicals in acidic media; pH < 5), are detected in TiO2 only dispersions; signals of O2*- are much weaker in the TiO2/ POM composite system under otherwise identical conditions. Experimental results infers that enhancement of charge separation in TiO2 photocatalysis does not always result in improvement of the efficiency of mineralization of organic substrates, and the reaction between organic radical cations and the formed superoxide radical anions may be responsible forthe mineralization of the chlorophenol. PMID- 14740756 TI - Interactions of soil-derived dissolved organic matter with phenol in peroxidase catalyzed oxidative coupling reactions. AB - The influence of dissolved soil organic matter (DSOM) derived from three geosorbents of different chemical composition and diagenetic history on the horseradish peroxidase (HRP) catalyzed oxidative coupling reactions of phenol was investigated. Phenol conversion and precipitate-product formation were measured, respectively, by HPLC and radiolabeled species analysis. Fourier transform infrared (FTIR) spectroscopy and capillary electrophoresis (CE) were used to characterize the products of enzymatic coupling, and the acute toxicities of the soluble products were determined by Microtox assay. Phenol conversion and precipitate formation were both significantly influenced by cross-coupling of phenol with dissolved organic matter, particularly in the cases of the more reactive and soluble DSOMs derived from two diagenetically "young" humic-type geosorbents. FTIR and CE characterizations indicate that enzymatic cross-coupling in these two cases leads to incorporation of phenol in DSOM macromolecules, yielding nontoxic soluble products. Conversely, cross-coupling appears to proceed in parallel with self-coupling in the presence of the relatively inert and more hydrophobic DSOM derived from a diagenetically "old" kerogen-type shale material. The products formed in this system have lower solubility and precipitate more readily, although their soluble forms tend to be more toxic than those formed by dominant cross-coupling reactions in the humic-type DSOM solutions. Several of the findings reported may be critically important with respect to feasibility evaluations and the engineering design of associated remediation schemes. PMID- 14740757 TI - Efficient hydrogen sulfide adsorbents obtained by pyrolysis of sewage sludge derived fertilizer modified with spent mineral oil. AB - Terrene, sewage sludge derived granulated fertilizer, was impregnated with spent mineral oil and then pyrolyzed at 600, 800, and 950 degrees C. Materials obtained were characterized from the point of view of the pore structure and surface chemistry. Then the H2S breakthrough capacitywas measured using a lab designed test. The results showed that the new adsorbents over perform by 30% materials obtained by simple thermal treatment of Terrene and by 230% virgin coconut shell based activated carbon. The surface reaction products were evaluated using thermal analysis. On the surface of new adsorbents hydrogen sulfide is oxidized mainly to elemental sulfur which is then deposited within the pore system. The breakthrough occurs when all small pores available to promote catalytic oxidation (caused by the inorganic sludge component) are filled with sulfur. An increase in pyrolysis temperature leads to an improvement in the performance of materials as hydrogen sulfide adsorbents. This is caused likely by changes in an inorganic phase and inorganic/carbonaceous phase interactions during pyrolysis. PMID- 14740758 TI - Treatment and prevention of equine gastric ulcer syndrome. AB - EGUS is a common problem in horses and foals. Acids are the important causative factors and current therapy targets the suppression of gastric HCl and creation of a permissive environment for ulcer healing. Diagnosis is based on history, clinical signs, gastroscopy, and response to treatment. Of the products available, only GastroGard (FDA approved) and ranitidine have been shown to be efficacious in the treatment of EGUS. Ranitidine is often associated with treatment failure as a result of incorrect dosing and lack of owner compliance, because of the three times daily dosing required. Also, EGUS occurs in critically ill neonatal foals, but the pathogenesis may be different than in adult horses and acid-suppressive therapy may not be as effective. PMID- 14740759 TI - Gastrointestinal protectants and cathartics. AB - The purpose of this article is to provide the reader with an overview of gastrointestinal cathartics and protectants and to point out possible applications for use in the horse with gastrointestinal disease. Most of the treatments described in this article have been used by the authors with apparent success; however, controlled studies with subsequent publication in the scientific literature with respect to these treatments in the horse are, for the most part, lacking. The authors view this emerging field of treatment as exciting and look forward to substantiating the efficacy of several of the treatments discussed. PMID- 14740760 TI - Nutrition for critical gastrointestinal illness: feeding horses with diarrhea or colic. AB - Horses with GI diseases such as colic and diarrhea are often intolerant of adequate enteral nutrition. Nutritional intervention should be an early part of therapeutic management in such cases. Protein and energy malnutrition in critically ill horses can have deleterious effects, including poor wound or incisional healing, reduced immunity, and weight loss. Early enteral or parenteral support should be provided to supply resting DE requirements in the equine ICU. PMID- 14740761 TI - Antimicrobial therapy for gastrointestinal diseases. AB - Antibiotics will always be needed in horses for many types of infections, but the adverse consequences also must be considered. For the conditions described in this article, there is justification for antibiotic therapy. The intestinal problems that antibiotics can induce are among the risks from their administration to horses. Disruption of the endogenous bacterial population, colitis, and diarrhea are the most common complications from antibiotic therapy. PMID- 14740762 TI - Fluid therapy in horses with gastrointestinal disease. AB - Fluid therapy is essential to the successful management of horses with gastrointestinal disease. Affected horses can present in a wide spectrum of metabolic derangement depending on the extent and severity of the underlying disease process. Precise quantitation of fluid deficits and losses is usually not possible; thus, formulating a rational plan, applying diligent and ongoing monitoring, and making adjustments to meet individual demands provide the best approach to fluid therapy in these horses. Although restoration of fluid and electrolyte homeostasis can be complicated, it is probably best in most cases to keep the approach as simple as possible and resist the urge to "over" manage. PMID- 14740763 TI - Treatment of endotoxemia. AB - Although recognition of clinical signs of endotoxemia is not difficult, determining the underlying cause for the development of endotoxemia may be a challenging task. For this reason, importance must be placed on performance of a thorough physical examination, attention to detail, and determination of whether or not surgical intervention is required. In most instances, a definitive diagnosis may require repeated examinations and laboratory work. In the meantime, affected animals require treatment for endotoxemia, with the aim being to reduce the duration of endotoxemia, prevent the interaction of additional endotoxins with cell surface receptors, and reduce the synthesis and release of inflammatory mediators. PMID- 14740764 TI - Treatment of acute and chronic gastrointestinal inflammation. AB - Treating inflammation in the equine gastrointestinal tract remains a challenge. Our most potent anti-inflammatory drugs, COX inhibitors and glucocorticoids, have unwanted effects on the gastrointestinal tract and host defense that often limit their use. Newer strategies targeting specific cells and molecules that regulate a subset of the events occurring during inflammation are rapidly becoming available and should allow clinicians to reduce the detrimental effects of inflammation without inhibiting the beneficial aspects. PMID- 14740765 TI - Treatment of gastrointestinal ischemic injury. AB - Ischemic injury is one of the most important causes of mortality in equine veterinary medicine. Although treatment of reperfusion injury has been attempted in a number of experimental trials to reduce the level of injury subsequent to an ischemic episode, this research has not resulted in the development of useful clinical treatments. Nevertheless, recent studies assessing intraluminal application of solutions containing antioxidants, nutrients, and vasodilators are promising. Furthermore, focusing on improving mucosal recovery after an ischemic event may provide an alternative method of reducing mortality. Potential treatments include administration of basement membrane components like hyaluronic acid, gut-specific nutrients like glutamine, and early return to feeding to stimulate endogenous repair mechanisms. Finally, recent studies evaluating NSAIDs have revealed the potential of flunixin meglumine to retard the mucosal repair process, indicating the need for judicious use of this drug. PMID- 14740766 TI - Clinical application of prokinetics. AB - Intestinal stasis or ileus is a significant cause of mortality and morbidity in horses and has been attributed to a variety of causes, including loss of intrinsic or extrinsic electrical activity, incoordination of contractile activity from regional stimuli, and dissociation between electrical and mechanical activity. Proposed mechanisms include systemic shock, electrolyte disturbances, persistent luminal distention, ischemia, inflammation, peritonitis, endotoxemia, and anesthesia. Because the cause of ileus is likely multifactorial, a variety of pharmaceutics have been used to target specific causes. Prokinetics are defined as agents that facilitate or enhance the net movement of feed material down the length of the intestinal tract and do not simply produce an uncoordinated increase in local contractile activity. The primary objective of pharmaceutic intervention is to augment the pathways that stimulate motility or attenuate the inhibitory neurons that predominantly suppress activity. The objective of this article is to summarize the actions of prokinetic agents available and suggest clinical applications. PMID- 14740767 TI - Prevention and treatment of gastrointestinal adhesions. AB - As with many aspects of clinical medicine, there is yet to be a single or definitive cure for postoperative adhesion formation. Current methods of prevention target risk factors predisposing horses to adhesion formation. Systemic pharmacologic therapies, such as antimicrobials, nonsteroidal anti inflammatory drugs, Salmonella antiserum, and hyperimmune plasma, help to reduce abdominal inflammation and minimize the effects of endotoxemia. Intra-abdominal or systemic heparin aids in enhancing peritoneal fibrinolysis. Prokinetic therapy promotes early postoperative return of intestinal motility, minimizing the propensity for adhesion formation between apposing adynamic segments of intestine. Mechanical separation of potentially adhesiogenic serosal and peritoneal surfaces is commonly achieved with use of abdominal lavage, protective coating solutions, and barrier membranes. Ongoing and future research is directed toward a better understanding of the local effects of intestinal trauma and the corresponding response of the fibrinolytic system. Recognition of horses at high risk for adhesion formation helps to guide the equine surgeon to an appropriate perioperative and intraoperative plan for adhesion prevention, including good surgical technique and a combination of adjunct therapies. PMID- 14740768 TI - Treatment of peritonitis. AB - In summary, peritonitis in the horse is a potentially life-threatening disease that must be treated promptly and aggressively. Therapy should be aimed at reducing systemic shock and hypovolemia, correction of the primary cause, antibiotic and anti-inflammatory therapy, and abdominal drainage and lavage. The prognosis depends on the ability to diagnose and treat the underlying cause and prevent the development of complications. Mortality rates can be as high as 59.7%, with horses developing postoperative peritonitis having a 56% mortality rate. Long-term complications like adhesion formation or internal abscesses may further reduce the survival rate. The prognosis is best determined by an early and quick response to aggressive treatment. PMID- 14740769 TI - Prevention and treatment of thrombosis, phlebitis, and laminitis in horses with gastrointestinal diseases. AB - Organ thrombosis and laminitis are life-threatening complications in horses with acute gastrointestinal disease, especially those diseases that cause disruption of the protective mucosal barrier. Prevention of these complications should be a high priority when treating horses with gastrointestinal diseases because even with proper and intensive treatments, laminitis or organ thrombosis may not be curative. Preventative therapy should include expedient and appropriate treatment of the primary disease, normalization of tissue perfusion and oxygenation, and inhibition of gut-derived toxins or their systemically activated biologic products along with i.v. replacement of depleted anticoagulant proteins. Additionally, several risk factors for thrombophlebitis in horses with intestinal disease have been identified, and those risk factors should be minimized during the illness. PMID- 14740770 TI - Treatment and prevention of intestinal parasite-associated disease. AB - Since 1917, only 11 new endoparasiticides have been developed for the horse, of which five chemical classes are in common use. The selection pressure of frequent administration of deworming doses for parasite control programs has been associated with the development of resistance of small strongyle parasites to the effects of benzimidazoles and pyrantel salts. Against the background of the inevitability of the occurrence of ivermectin/moxidectin resistance, responsible use of equine anthelmintics based on the clinical pharmacology of the compounds and the biology/epidemiology of intestinal parasites is a major issue for equine clinicians. The evidence base for the recommendations for treatment and control of equine intestinal parasites is sparse, however, and few robust data exist from controlled clinical trials to validate current guidelines. PMID- 14740771 TI - How common is hypertension in young adults in Singapore? PMID- 14740772 TI - William Harvey (1578-1657): discoverer of the circulation. PMID- 14740773 TI - Hypertension in young adults--an under-estimated problem. AB - AIM: To study the prevalence of hypertension and "white coat hypertension" in young adult Asian males, and identify the associated risk factors. METHODS: Population-based descriptive analysis of 3,352 Singapore military conscripts presenting consecutively for medical screening, followed by case-control study of subjects with elevated blood pressure. A standard protocol for assessing elevated blood pressure, 24-hour ambulatory monitoring and detailed interviews were performed. Main study outcomes are prevalence rate of hypertension and "white coat hypertension", mean blood pressure readings, and adjusted odds ratios for associated variables. RESULTS: Prevalence of hypertension and "white coat hypertension" was 1.6% (95% CI 1.2, 2.0) and 2.0% (95% CI 1.5, 2.5) respectively. Twenty-four-hour ambulatory monitoring was required to differentiate the two conditions, with a fall of 22.5 mmHg (95% CI 19.7, 25.3) observed between first visit and day-time ambulatory mean systolic blood pressures. There was strong association between hypertension and obesity (adjusted odds ratio using Body Mass Index: 1.19, p<0.001). Other important variables included parental history of hypertension, Malay ethnicity and low socio-economic status, although there was no significant correlation in our regression model. CONCLUSION: This study provides population-based data on hypertension in young Asian adults. While the prevalence of hypertension is low compared to older age groups, it remains important to detect cases early, as appropriate treatment may mitigate long-term cardiovascular risks and reduce target organ damage. There is a clear role for ambulatory blood pressure monitoring for differentiating true hypertension from "white coat hypertension". There may be a role for targeted screening of high risk groups, particularly the obese. PMID- 14740774 TI - Childhood acute pancreatitis in a children's hospital. AB - OBJECTIVE: To analyse the cases of acute pancreatitis presented to a children's hospital in Singapore. METHODS: Clinical charts of all children, aged under 18 years, who presented to our hospital for the first time with pancreatitis (ICD search criteria = 577.x) between the period of 1998 and mid-2002 were reviewed. Parameters analysed included presenting features, aetiology of the acute pancreatitis, length of hospital stay, complications, treatment and outcome. RESULTS: There were 12 cases in the review period, and the attributable causes in these cases were, in descending order, trauma, drug-induced, anatomical anomalies, poisoning and idiopathic. Of interest were two patients whose pancreatitis were results of child abuse. The most common symptoms were abdominal pain (n=11) and vomiting (n=7), though only five patients localised the pain to the epigastrium. Abdominal tenderness could be elicited in all the patients. Eleven had evidence of acute pancreatitis from computerised tomography (CT) whilst the twelfth was diagnosed with ultrasonography. The peak amylase levels amongst these patients were not high, with a median of 512.5 U/L. In the acute stage, only one patient required operative intervention whilst the remainder were managed conservatively. The mean length of hospital stay was 12.41 +/- 4.54 days. The complications encountered included pseudocyst formation, ascites, hypocalcaemia, pleural effusion and coagulopathy. CONCLUSIONS: The diagnosis of acute pancreatitis in children can be difficult. This is often due to ambiguous symptoms, signs and laboratory results. CT and ultrasound are essential investigations in the diagnosis and subsequent follow-up. PMID- 14740775 TI - War with SARS: an empirical study of knowledge of SARS transmission and effects of SARS on work and the organisations. AB - AIM OF STUDY: This study examined the level of knowledge of SARS transmission among Singaporeans and their willingness to disclose their health condition to others. We also examined respondents' perceived effects of SARS on work and organisations and their attitudes toward issues of privacy and disclosure of medical information. METHOD: Respondents comprised MBAs (Master of Business Administration students) and human resource managers who attended classes in a local tertiary institution. Data were collected via an email survey. A total of 101 completed surveys were received and included in data analyses. RESULTS: Results suggest that despite rather intensive efforts to generate awareness about SARS transmission, a certain level of uncertainty about how SARS can be transmitted still prevails. This is not surprising, given that SARS is a relatively new medical problem. Our findings also suggest that while respondents unanimously agreed that they would inform their parents, spouse, siblings and employers if they were tested positive for SARS, they were more ambivalent about disclosing such information to their neighbours and colleagues. Findings also suggest that having a SARS or probable SARS case in the company would disrupt the flow of work and affect employees' morale. CONCLUSION: Results of this study have significant implications for efforts to educate Singaporeans about the disease and the management of SARS at the workplace. PMID- 14740776 TI - Neuraxial block for labour analgesia--is the combined spinal epidural (CSE) modality a good alternative to conventional epidural analgesia? AB - AIM: Apart from conventional epidural analgesia (EA), the combined spinal epidural (CSE) modality is fast becoming a popular technique for treating labour pain. In this study, we investigated the differences in the patient profile and outcome between CSE and EA for labour pain in KK Women's and Children's Hospital. METHODOLOGY: Data pertaining to 1,532 healthy parturients who had received either CSE or EA for labour pain during a six-month period was systematically collected by using a specially designed form. Multiple logistic regression analysis was used to determine the independent predictors of patient satisfaction and the relation of parturient factors on the choice of block. The side effects and the outcome of labour were also compared. RESULTS: CSE accounted for 80% of all neuraxial blocks performed for labour analgesia (vs 20% for EA). Anaesthesiologists were more inclined to using CSE than EA for multiparous parturients (OR 2.03, p<0.01) in a more painful (OR=1.61, p=0.03) and advanced stage of labour (OR=1.12, p=0.03). The need for supplemental analgesics was greater for EA (p<0.01). Patient satisfaction was higher for CSE (OR=1.77, p<0.026). CSE had a higher risk of pruritus (29% vs 14%, p<0.01) but lower risk of post block neural deficits (0% vs 2%, p<0.01) than EA. No difference in the mode of delivery was detected between the two groups. CONCLUSION: CSE is a safe and good alternative to EA as a technique of neuraxial block for labour analgesia. PMID- 14740777 TI - Advanced disseminated gastric carcinoma in pregnancy. AB - Gastric cancers associated with pregnancy are rare and often diagnosed at advanced stages where curative therapies are not possible. The outcomes have generally been very poor with death occurring within months of diagnosis. Suspicion and early endoscopy are necessary for early diagnosis. We herein report a case of advanced disseminated gastric cancer presenting in the third trimester with pre-eclampsia and death occurring less than a month after diagnosis. PMID- 14740778 TI - Acute interstitial nephritis in Singapore: a report of five cases. AB - Interstitial nephritis is an uncommon cause of acute renal failure. Reported incidence varies widely in the literature and may depend on several factors i.e. geographical location, diagnostic criteria, dietary, environmental factors and therapeutic practices. This is a retrospective study of biopsy proven interstitial nephritis in National University Hospital Singapore. We report five cases out of a total of 349 biopsies carried out during a five-year period between September 1997 and August 2002. Four patients presented acutely with fever and or cough. In four patients, there was exposure to traditional Chinese medications and/or drugs. Renal failure in four out of the five patients progressed rapidly, three of whom required dialysis. One patient was treated with steroids. Renal function recovered in all patients with one patient who had significant residual renal impairment after one month. PMID- 14740779 TI - Restenosis in percutaneous coronary intervention--is drug-eluting stent the answer? AB - The long-term success of percutaneous coronary intervention in the treatment of coronary artery disease is hampered by the occurrence of restenosis, which often necessitates repeat hospitalisations or coronary interventions. The advent of drug-eluting stents, particularly those coated with sirolimus and paclitaxel, may be the breakthrough in the battle against restenosis that interventional cardiologists have been waiting for, and we review the currently available evidence for this. Despite the growing enthusiasm, we should not forget that this new technology is still in its relative infancy, and there remain many unanswered questions, particularly about the long-term effect of using these stents. PMID- 14740780 TI - Article: Cerebrotendinous Xanthomatosis in three siblings from a Chinese family. PMID- 14740781 TI - Clinics in diagnostic imaging (89). Infantile hepatosplenic haemangioendotheliomas. AB - The ultrasonographical and computed tomography findings of a six-month-old female infant with haemangioendotheliomas of the liver and spleen are described. The splenic lesion had an unusual ultrasonographical appearance of a well-defined isoechoic mass with a hypoechoic rim. Diagnosis was confirmed by histological examination of the hepatic biopsy specimen. Hepatosplenic lesions in the first year of life may be due to a variety of pathological processes. It is important to include haemangioendotheliomas in the differential diagnosis of hepatosplenic masses in an infant. PMID- 14740782 TI - Tachycardia-induced heart failure does not alter myocardial P-glycoprotein expression. AB - STUDY OBJECTIVE: To determine the effects of tachycardia-induced heart failure on myocardial P-glycoprotein (P-gp) expression. DESIGN: Nonblinded, parallel, sham controlled, animal model study. SETTING: University laboratory. ANIMALS: Thirty mongrel dogs. INTERVENTION: Heart failure was induced by rapid ventricular pacing over 4 weeks; sham procedures were performed for the control group. MEASUREMENTS AND MAIN RESULTS: Myocardial biopsies were taken from the left ventricular lateral wall and prepared for P-gp quantification by laser-induced fluorescence. The relative amount of P-gp messenger RNA (mRNA) was assessed by reverse transcriptase polymerase chain reaction. Rapid ventricular pacing produced heart failure and reduced the area ejection fraction from 48% +/- 6% to 21% +/- 6% (p<0.05 vs baseline). However, heart failure did not alter the quantity of myocardial P-gp (0.20 +/- 0.02 microg/ml for the control group vs 0.23 +/- 0.02 microg/ml for the intervention group, p=0.4). Furthermore, heart failure did not alter P-gp expression significantly. CONCLUSION: Myocardial P-gp does not change in response to tachycardia-induced heart failure. Thus, there is a low likelihood for P-gp-related drug resistance during a syndrome similar to tachycardia-induced heart failure. PMID- 14740783 TI - Pharmacodynamics of meropenem and imipenem against Enterobacteriaceae, Acinetobacter baumannii, and Pseudomonas aeruginosa. AB - STUDY OBJECTIVE: To compare the pharmacodynamics of meropenem and imipenem, both administered as 500 mg every 6 hours, against populations of Enterobacteriaceae, Acinetobacter baumannii, and Pseudomonas aeruginosa. DESIGN: Ten thousand-subject Monte Carlo simulation. INTERVENTION: Variability in total body clearance (ClT), volume of distribution as calculated by the terminal elimination rate (Vdbeta), and minimum inhibitory concentration (MIC) distributions (Escherichia coli, Klebsiella pneumoniae, Enterobacter cloacae, Serratia marcescens, A. baumannii, P. aeruginosa) were derived from the literature for both meropenem and imipenem. For the free drug concentrations, the percentage of the dosing interval that the drug concentrations remain above the MIC (%T>MIC) for each carbapenem-bacteria combination was calculated for 10,000 iterations, substituting a different ClT, Vdbeta, fraction of unbound drug, and MIC into the equation each time based on the probability distribution for each parameter. Probabilities of attaining targets of 30%, 50%, and 100% T>MIC were calculated. MEASUREMENTS AND MAIN RESULTS: Meropenem free drug %T>MIC exposure was significantly greater than that of imipenem against Enterobacteriaceae and P. aeruginosa, whereas imipenem exposure was greater for A. baumannii. For both agents, free drug %T>MIC exposure was greatest against Enterobacteriaceae and less for A. baumannii and P. aeruginosa. Probabilities of target attainment for 30% and 50% T>MIC were similar between drugs for most bacteria. At 100% T>MIC, meropenem target attainments were greater than those of imipenem against Enterobacteriaceae and P. aeruginosa, and imipenem attainment was higher for A. baumannii. CONCLUSION: The probability of attaining lower pharmacodynamic targets for most gram-negative bacteria is similar for these carbapenems; however, differences become apparent as the pharmacodynamic requirement increases. Further study of the benefits of achieving this pharmacodynamic breakpoint with a higher probability of attaining targets is necessary. PMID- 14740784 TI - Temozolomide in patients with advanced cancer: phase I and pharmacokinetic study. AB - STUDY OBJECTIVE: To determine the maximum tolerated dose, dose-limiting toxicity, pharmacokinetics, and potential antitumor activity of temozolomide administered as a single dose every 28 days. DESIGN: Open label, phase I, dose-escalation trial. SETTING: University-affiliated cancer center. PATIENTS: Eleven patients aged 33-73 years with a documented solid tumor or lymphoma who failed therapy of proven efficacy for their disease or had a disease for which no conventional therapy was available. INTERVENTION: Temozolomide 500 mg/m2 was administered as a single oral dose every 28 days. Doses were escalated to 750 or 1000 mg/m2. No intrapatient dose escalation was allowed. At least two patients were enrolled at each dose level. Patients who did not have progressive disease and did not experience a dose-limiting toxicity, or experienced a dose-limiting toxicity but were eligible for dose reduction, were eligible to continue on the study. MEASUREMENTS AND MAIN RESULTS: Pharmacokinetic analysis was performed for temozolomide and its active metabolite, 5-(3-methyltriazeno)-imidazole-4 carboxamide (MTIC). Neutropenia and thrombocytopenia were dose limiting at 1000 mg/m2. Temozolomide was absorbed rapidly (mean time to maximum concentration 1.4 hrs) and eliminated, with average half-life and apparent oral systemic clearance values of 1.8 hours and 97 ml/minute/m2, respectively. Mean systemic exposure to MTIC was 3.7% of temozolomide. No objective responses were observed. The maximum tolerated dose of temozolomide was 750 mg/m2. CONCLUSION: Temozolomide 750 mg/m2 administered orally every 28 days was well tolerated. Alternate temozolomide dosing schedules such as continuous daily administration may enhance antitumor activity through sustained depletion of the DNA repair protein O6-alkylguanine DNA alkyltransferase. PMID- 14740785 TI - Effect of fluticasone propionate nasal spray on bioavailability of intranasal hydromorphone hydrochloride in patients with allergic rhinitis. AB - STUDY OBJECTIVE: To investigate the effect of the nasal corticosteroid fluticasone propionate on the bioavailability and pharmacokinetics of single-dose intranasal hydromorphone hydrochloride in patients with allergic rhinitis. DESIGN: Randomized, three-way, crossover pharmacokinetic study. SETTING: University clinical research unit. PATIENTS: Twelve patients with allergic rhinitis. INTERVENTION: Hydromorphone hydrochloride 2.0 mg was administered by intravenous infusion (treatment A), intranasal spray without allergic rhinitis treatment (treatment B), and intranasal spray after 6 days of fluticasone propionate (treatment C). Blood samples were collected serially from 0-16 hours. MEASUREMENTS AND MAIN RESULTS: Pharmacokinetic parameters were determined by noncompartmental methods. An analysis of variance (ANOVA) model was used for statistical analysis. Mean (% coefficient of variation) absolute bioavailability of intranasal hydromorphone was 51.9% (28.2) and 46.9% (30.3) in patients with allergic rhinitis with and without treatment with fluticasone propionate, respectively. Mean maximum concentration (Cmax) values were 3.02 and 3.56 ng/ml, respectively. No statistical differences in Cmax and area under the concentration versus time curve were detected between intranasal treatments. Bioavailability values for both intranasal treatments were lower than those in healthy volunteers (57%). Median time to Cmax (Tmax) values were significantly different (p=0.02) for treatments B and C (15 and 30 min, respectively) using rank-transformed Tmax for ANOVA. Adverse effects were consistent with known effects of hydromorphone administered by other routes, with the exception of bad taste after intranasal administration. CONCLUSION: Hydromorphone was rapidly absorbed after nasal administration, with maximum concentrations occurring for most subjects within 30 minutes. Allergic rhinitis may affect pain management strategies for intranasal hydromorphone, with a delay in onset of action for patients treated with fluticasone propionate. PMID- 14740786 TI - Acute and continuation treatment adequacy with venlafaxine extended release compared with fluoxetine. AB - STUDY OBJECTIVE: To compare treatment adequacy in the management of depression during the acute and continuation phases between patients newly treated with venlafaxine extended release (XR) and those newly treated with fluoxetine. DESIGN: Retrospective observational analysis of pharmacy claims data. SETTING: Large California-based managed care organization. PATIENTS: A total of 11,298 patients newly prescribed venlafaxine XR or fluoxetine between January 1, 2000, and February 28, 2001, and continuously enrolled throughout the study, as well as a subset of 7430 patients who continued taking venlafaxine XR or fluoxetine during the follow-up period. MEASUREMENTS AND MAIN RESULTS: The Health Plan Employer Data and Information Set definition was used for continuous antidepressant treatment during the acute and continuation phases. Treatment adequacy was determined for those deemed continuous. Patients receiving within +/ 10% of the target dose for each drug (venlafaxine XR 75-150 mg, fluoxetine 20 mg) were defined as receiving an adequate dose. Logistic regression was used to evaluate venlafaxine XR versus fluoxetine on treatment adequacy, controlling for age, sex, physician specialty, and pharmacy benefit. The unadjusted adequacy rate for the venlafaxine XR-only group was 79% versus 57% for the fluoxetine-only group for 84 continuous days (p<0.0001) and 77% versus 52%, respectively, for 180 continuous days (p<0.0001). The adjusted odds ratios of achieving treatment adequacy with venlafaxine XR only versus that with fluoxetine only were 3.05 (95% confidence interval [CI] 2.65-3.52) for 84 continuous days and 3.57 (95% CI 3.00 4.24) for 180 continuous days. CONCLUSION: Patients newly prescribed venlafaxine XR were at least 3 times more likely to achieve treatment adequacy for 84 and 180 days compared with those newly prescribed fluoxetine. Treatment adequacy as a proxy for optimal treatment may be an important factor to consider when selecting an antidepressant drug. PMID- 14740787 TI - Daptomycin. AB - Daptomycin, the first in a class of agents known as lipopeptides, is a novel antimicrobial agent used for the treatment of gram-positive infections. The compound has a distinctive mechanism of action that exerts its bactericidal activity by disrupting plasma membrane function without penetrating into the cytoplasm. The agent has received much interest because of its activity against multidrug-resistant, gram-positive bacteria such as methicillin-resistant Staphylococcus aureus, vancomycin-resistant enterococci, and glycopeptide intermediate and -resistant S. aureus. Daptomycin demonstrates concentration dependent killing and is eliminated primarily by glomerular filtration. It was approved in September 2003 for the treatment of complicated skin and soft tissue infections. It has a safety profile similar to other agents commonly administered to treat gram-positive infections. Daptomycin is a welcome addition to the antimicrobial armamentarium for the treatment of bacterial infections. Further clinical experience with this compound will help define its role in the treatment of resistant gram-positive organisms. PMID- 14740788 TI - Oritavancin and tigecycline: investigational antimicrobials for multidrug resistant bacteria. AB - The advent of multidrug-resistant gram-positive aerobes such as Staphylococcus aureus, Streptococcus pneumoniae, and the enterococci, which are resistant to beta-lactams, vancomycin, and a host of other commonly used antimicrobials, has complicated our approach to antibiotic therapy. Despite marketing of the first oxazolidinone, linezolid, and the streptogramin combination, quinupristin dalfopristin, an urgent need exists for more agents to combat these pathogens. Two such agents, the glycopeptide oritavancin (LY333328) and the glycylcycline tigecycline (GAR-936), are in phase III clinical trials. These agents, which require parenteral administration, exhibit substantial in vitro activity against a variety of gram-positive aerobes and anaerobes, including the multidrug resistant organisms listed previously. Only tigecycline demonstrates useful activity against gram-negative organisms. Combination therapy of these agents with ampicillin or aminoglycosides frequently leads to synergistic in vitro activity against multidrug-resistant staphylococci and streptococci. These agents are also active in a variety of animal models of systemic and localized infections. Few published efficacy and tolerability data are available in humans. If controlled clinical trial data verify these agents' efficacy and tolerability, both drugs should become welcome additions to the available antimicrobials. However, restricting their use to the treatment of infections caused by bacteria resistant to other antimicrobials, especially multidrug-resistant staphylococci and streptococci, may prolong their clinical utility by retarding the development of resistance. Careful surveillance of bacterial sensitivity to these agents should be undertaken to assist clinicians in the decision whether or not to use these agents empirically to treat infections caused by suspected multidrug resistant gram-positive pathogens. PMID- 14740789 TI - Poppers: epidemiology and clinical management of inhaled nitrite abuse. AB - Commonly referred to as "poppers," inhaled nitrites have a long history of abuse. Poppers are rapid-onset, short-acting potent vasodilators that produce a rush characterized by warm sensations and feelings of dizziness. Poppers sometimes are used to facilitate anal intercourse because of their actions on the anal sphincter. Epidemiologically, the frequent use of nitrites by men who have sex with men has led some experts to implicate these chemicals in the pathogenesis of Kaposi's sarcoma and acquired immunodeficiency syndrome. Controlled clinical trials to examine this potential correlation have not been conducted, and the use of nitrites simply may be a marker for other high-risk behaviors such as unprotected sex. Although regulated in the United States, many nitrite compounds and isomers are sold at various venues including bars, bookstores, and over the Internet. Adverse effects associated with these products vary from mild allergic reactions to life-threatening methemoglobinemia. The potential for drug-drug interactions and a propensity toward unsafe sex also exist. Clinicians should be familiar with the populations most likely to abuse these agents and with the clinical effects and management guidelines for acute ingestions. PMID- 14740790 TI - Nonhormonal alternatives for the treatment of hot flashes. AB - OBJECTIVE: To review the literature on clonidine, venlafaxine, selective serotonin reuptake inhibitors, and gabapentin for the treatment of hot flashes. DATA SOURCES: A MEDLINE search (January 1966-July 2003) was conducted to identify English-language literature available on the treatment of hot flashes that focused on clonidine, venlafaxine, selective serotonin reuptake inhibitors, and gabapentin. These articles, relevant abstracts, and additional references listed in articles were used to collect pertinent data. STUDY SELECTION: All controlled and uncontrolled trials were reviewed. DATA SYNTHESIS: In women unable or unwilling to take hormonal therapies, several nonhormonal alternatives have been evaluated in small controlled and uncontrolled trials. Oral and transdermal formulations of clonidine are moderately effective in reducing hot flashes. Results of studies evaluating venlafaxine, paroxetine, and gabapentin suggest greater reductions in hot-flash frequency and severity compared with those of clonidine. Fluoxetine appears to display a modest benefit compared with paroxetine, although no comparative trials have been conducted. Most women studied in these trials had a history of breast cancer, and many were taking concurrent tamoxifen. All of these agents were fairly well tolerated. CONCLUSIONS: Clonidine, venlafaxine, paroxetine, fluoxetine, and gabapentin are nonhormonal agents that have demonstrated efficacy in small controlled and uncontrolled trials in reducing hot flashes and should be considered in patients unwilling or unable to take hormonal therapies. PMID- 14740791 TI - Evaluation of carvedilol for the treatment of portal hypertension. AB - Development of bleeding gastroesophageal varices is a serious consequence of portal hypertension secondary to cirrhosis. Nonselective beta-blockers have been used to reduce portal pressures and prevent primary and secondary bleeding episodes. However, up to two thirds of patients may not respond appropriately to these agents. Nonselective beta-blockers combined with vasodilatory drugs result in enhanced lowering of portal pressures by targeting several mechanisms involved in this process. Unfortunately, this practice is associated with increased adverse effects, such as hypotension, and minimal reductions in mortality. Carvedilol possesses both nonselective beta-antagonist and alpha1-receptor antagonist activity. Given its combined mechanism of action, carvedilol presents a potential option for lowering portal pressures. Its effects on lowering portal pressures and its role in therapy are undefined. Using MEDLINE (1966-2003) and International Pharmaceutical Abstracts (1970-2003), the English-language literature was searched to identify human studies assessing carvedilol's effects on lowering portal pressure. In general, carvedilol therapy was associated with mean reductions of 16-43% in portal pressure, assessed by the hepatic venous pressure gradient (HVPG) after single and multiple doses. Studies comparing carvedilol with propranolol revealed equal or enhanced efficacy in lowering HVPG. Large percentages of patients had significant HVPG reductions to levels that prevent variceal bleeding. Carvedilol also was associated with substantial symptomatic hypotension, especially in patients with ascites or Child-Pugh class B or C cirrhosis. Efficacy and adverse effects generally seem to be dose related. Carvedilol appears to be a potentially viable option for treating portal hypertension. Further multiple-dose trials comparing carvedilol with standard therapy are needed to assess the agent's long-term safety and effectiveness in preventing variceal bleeding. PMID- 14740792 TI - Key articles and guidelines relative to treatment of patients with acute coronary syndromes. AB - Patients with cardiovascular disease who have acute coronary syndromes (ACS) are at risk of significant morbidity and mortality. Also, treatment of these patients in the early-phase setting has a significant financial impact on the health care system. With the existence of numerous pharmacologic agents, abundance of major clinical trials, and several nationally recognized clinical guidelines, compiling the needed reference material to make evidence-based decisions on the care of patients with ACS can be difficult for clinicians. To assist clinicians in this endeavor, we complied pertinent articles and guidelines that have shaped the current treatment of patients with ACS. Owing to the rapidly evolving body of evidence in the management of ACS, this compilation will require periodic updating. PMID- 14740793 TI - Evaluation of kidney function in patients with acute renal failure using high performance liquid chromatography: a case report. AB - Accurate measurement of glomerular function rate (GFR) in the setting of acute renal failure is difficult to achieve with current clinical methods, such as measuring plasma creatinine concentration and 24-hour urine creatinine clearance. High-performance liquid chromatography was used to measure GFR directly in a critically ill patient with acute renal failure. This approach involved evaluating the elimination kinetics of nonionic contrast material administered intravenously for radiologic imaging. It required no additional patient exposure to radiographic contrast media and enabled caregivers to determine kidney function accurately in the presence of worsening clinical status and delayed changes in plasma creatinine. This and other methods for more accurate measurement of GFR in patients with acute renal failure may provide the foundation for clinical studies that assess the severity and management of acute renal failure. PMID- 14740794 TI - Liver function test abnormalities and pruritus in a patient treated with atorvastatin: case report and review of the literature. AB - The 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors (also known as statins) are associated with elevated transaminase levels in 1-3% of patients. Therapy with these drugs requires monitoring of alanine aminotransferase (ALT) levels because animal studies and premarketing clinical trials showed signs of hepatotoxicity that were primarily minor elevations of ALT. Nevertheless, postmarketing experience suggests that hepatotoxicity is rare, and that elevated ALT levels are reversible with continued therapy and probably are related to cholesterol lowering. Based on the low occurrence of ALT elevations and the lack of clinical evidence of hepatotoxicity, some clinicians are calling for a change in the current practice of monitoring liver function tests. We report, however, the case of a 71-year-old woman who was receiving atorvastatin and experienced elevated transaminase levels on two occasions, and developed pruritus on rechallenge with the drug. Thus, clinicians should be aware of asymptomatic elevations in liver function tests in patients receiving atorvastatin who do not have known risk factors for liver damage. PMID- 14740795 TI - Amifostine-induced fever: case report and review of the literature. AB - Amifostine, also known as WR-2721 (S-2-(3-aminopropylamino)-ethylphosphorothioic acid), is an organic thiophosphate cytoprotective agent used to reduce the frequency of severe xerostomia in patients with cancer undergoing postoperative radiation of the head and neck. A 56-year-old Caucasian man who received concomitant chemotherapy and radiation for head and neck cancer developed fever concurrent with the administration of amifostine. To our knowledge, this is the first case report that demonstrates the occurrence of fever with low-dose amifostine therapy without the manifestation of accompanying rash or hypotension. Patients receiving amifostine who develop only fever should be evaluated for an adverse drug reaction, as well as for sepsis and fevers of neutropenia, and it may be necessary to discontinue the drug. Recognition of amifostine as the cause of this adverse event may prevent the cost and inconvenience of a hospital admission. PMID- 14740796 TI - Telemedicine and tele-ophthalmology: part II. PMID- 14740797 TI - Effectiveness and safety of screening for diabetic retinopathy with two nonmydriatic digital images compared with the seven standard stereoscopic photographic fields. AB - BACKGROUND: The use of nonmydriatic cameras, which offer ease of screening and 45 degrees immediate imaging of the fundus, is gaining increasing acceptance for screening programs tailored to diverse conditions. We performed a study to evaluate the effectiveness and safety of screening for diabetic retinopathy with two nonmydriatic camera images compared with the seven standard stereoscopic 30 degrees fields (7SF). We also wished to determine whether safe screening guidelines could be established to identify patients needing referral to an ophthalmologist. METHODS: In this prospective masked cross-sectional study, we evaluated agreement in the assessment of the severity of diabetic retinopathy by means of two 45 degrees images centred on the optic disc and on the macula obtained with the Topcon CRW6 nonmydriatic camera and by means of 7SF photography and ophthalmologic slit-lamp biomicroscopy, both performed with pupil dilation. Between November 2000 and June 2001, 98 adult patients known to have type 1 or 2 diabetes mellitus who presented for the first time to the diabetic retinopathy clinic of a tertiary care centre in Montreal were enrolled consecutively. Thus, patient recruitment was weighted toward more severe retinopathy to ensure sufficient representation of less frequent but more severe levels. Each patient underwent nonmydriatic fundus photography of both eyes, followed by a complete ophthalmologic examination with pupil dilation by a single retina specialist and 7SF photography of both eyes with pupil dilation. The level of retinopathy was graded independently in each eye from the 7SF photographs according to the Early Treatment Diabetic Retinopathy Study (ETDRS) scale by two graders; an independent retina specialist adjudicated the rare instances of interreader disagreement in a masked fashion. Two months later, two graders independently graded the nonmydriatic images in a blinded fashion according to the ETDRS scale; a third observer adjudicated the rare instances of interreader disagreement. We measured concordance between grading results with the various screening techniques using the weighted and unweighted kappa statistic. We used sensitivity and specificity indices to determine safe screening guidelines to identify patients needing referral to an ophthalmologist. RESULTS: There was substantial agreement in the grading of retinopathy with nonmydriatic camera imaging and with 7SF photography, both for all eyes (kappa = 0.626 [standard deviation (SD) 0.045]) and for the eye with more severe disease (kappa = 0.654 [SD 0.063]). With nonmydriatic camera imaging, screening thresholds for patient referral to an ophthalmologist of very mild retinopathy (ETDRS grade 20), mild retinopathy (ETDRS grade 35) and moderate retinopathy (EDTRS grade 43) had sensitivity values of 97.9%, 97.1% and 53.3% respectively and specificity values of 81.3%, 95.5% and 96.9% respectively. Screening thresholds of very mild or mild retinopathy both correctly identified 100% of eyes with severe nonproliferative or proliferative retinopathy. With a screening threshold of mild retinopathy, screening with the nonmydriatic camera would lead to referral to an ophthalmologist of 37.8% of patients because of detected disease and of an additional 17.3% because of insufficient image quality in at least one eye, for a total of 55.1%. The overall sensitivity and specificity of a two-field nonmydriatic screening strategy with a threshold of mild retinopathy for referral of patients with insufficient image quality in at least one eye are 97.7% and 84.0% respectively. INTERPRETATION: Our results suggest that two-field nonmydriatic camera imaging is a safe screening strategy that may identify the patients with diabetes most in need of ophthalmologic care. PMID- 14740798 TI - Comparison of two, three and four 45 degrees image fields obtained with the Topcon CRW6 nonmydriatic camera for screening for diabetic retinopathy. AB - BACKGROUND: The use of the nonmydriatic camera is gaining increasing acceptance for the detection of diabetic retinopathy when integrated into a community tailored program. We performed a study to evaluate the optimal number and positioning of photographic fields necessary to screen for diabetic retinopathy with the Topcon CRW6 nonmydriatic camera. METHODS: In this prospective masked cross-sectional comparative study, we compared the assessment of diabetic retinopathy using two, three or four 45 degrees fundus images (centred respectively on the disc and the macula; on the disc, on the macula and temporal to the macula; and on the disc, on the macula, temporal to the macula and superotemporal to the macula, including the superior temporal vein) acquired with the Topcon CRW6 nonmydriatic camera, with the grading of the seven standard stereoscopic 30 degrees field photographs (7SF). The study population consisted of 98 consecutive adult patients known to have type 1 or 2 diabetes mellitus who presented for the first time to the diabetic retinopathy clinic of a tertiary care centre in Montreal. All patients underwent four nonmydriatic fundus photography sessions of both eyes, 7SF photography with pupil dilation and a complete ophthalmologic examination with pupil dilation by a retina specialist. RESULTS: Compared to the 7SF, the sensitivity and specificity of screening for any retinopathy (Early Treatment Diabetic Retinopathy Study [ETDRS] grade greater than 10) using the two central fields were 95.7% and 78.1% respectively. The corresponding values with three image fields were 97.6% and 71.9%, and with four image fields, 97.6% and 65.6%. The sensitivity and specificity of screening for mild or worse disease (ETDRS grade 35 or greater) using the two central fields were 87.5% and 92.3%. The corresponding values with three image fields were 88.9% and 94.6%, and with four image fields, 88.9% and 91.9%. Poor image quality occurring with the addition of extra fields resulted in an increase of 6.2% in the rate of referral to an ophthalmologist. The use of two image fields missed no cases of retinopathy. INTERPRETATION: The use of image fields in addition to the two 45 degrees images centred on the disc and the macula on imaging with the Topcon CRW6 nonmydriatic camera did not significantly increase the sensitivity or specificity of screening for diabetic retinopathy. Contrary to the desired effect, the addition of fields resulted in diminished utility and cost effectiveness of this screening approach. PMID- 14740799 TI - Use of telemedicine in screening for diabetic retinopathy. AB - BACKGROUND: A diabetic retinopathy screening program was introduced in September 2000 at a university-affiliated hospital in Montreal to meet the demands of an overburdened health care system. In this article we describe our initial experience with this program. METHODS: A Canon CR6-45NM nonmydriatic 45 degrees camera was installed in the outpatient endocrinology clinic of the hospital. All patients who visited the clinic between September 2000 and January 2001 agreed to participate in the program. For each patient, a short questionnaire (age, sex, type of diabetes mellitus, time since diagnosis, prior retinal laser therapy and presence of end-organ involvement) was filled in. Subsequently at least two photographs were obtained of each eye. The photographs were sent via the hospital intranet to a retinal specialist within the hospital. The specialist reviewed the images unchanged and sent a report (grading of the level of diabetic retinopathy, presence or absence of macular hard exudates, quality of the images obtained and follow-up recommendations) via the hospital intranet to the endocrinology service. If the images were judged to be of poor quality and could not be read by the retina specialist, the patient was asked to see his or her regular ophthalmologist. RESULTS: Images of 830 eyes of 415 patients (211 men and 204 women with a mean age of 57.5 years) were obtained. Most (83.6%) had type 2 diabetes. The average duration of disease was 13.3 years. Macular hard exudates were observed in 50 eyes (6.0%). Nonproliferative diabetic retinopathy was noted in 117 eyes (14.1%) and proliferative diabetic retinopathy in 15 eyes (1.8%). Unexpected findings included epiretinal membranes (in three eyes), macular holes (in two), bilateral cotton-wool spots (in one patient) and central retinal vein occlusion (in one eye). Overall, 10% of the patients were referred to a retina specialist based on the screening photographs. The main difficulty encountered with the screening program was image inconsistency. Overall, 35% of the images graded were felt to be of poor quality, the most common reason being poor exposure (84.4%). The quality of the images improved significantly over the study period (p < 0.01). Images obtained from patients aged 65 years or more were consistently poorer than those obtained from patients less than 65 years of age (p < 0.001). INTERPRETATION: Despite the imperfections of such a system, our experience was favourable, allowing us to screen large numbers of patients in a cost-effective, reliable manner. PMID- 14740800 TI - Comparison of outcomes of viscocanalostomy and phacoviscocanalostomy. AB - BACKGROUND: Many adults who undergo glaucoma filtering surgery also need surgical treatment of coexisting cataract. Recent reports of the use of new alternative non-penetrating techniques in combination with phacoemulsification have given encouraging results. The purpose of this study was to compare the safety and efficacy of viscocanalostomy and phacoviscocanalostomy in the management of medically uncontrolled open-angle glaucoma. METHODS: Twenty patients who underwent phacoviscocanalostomy in one eye each were enrolled in the study. Twenty matching patients who underwent viscocanalostomy in one eye each at our institution were selected as the control group. Postoperative reductions in intraocular pressure (IOP) and number of antiglaucoma medications were compared between the groups. Visual acuity and complications were secondary outcomes. RESULTS: Significant reductions of IOP and of use of antiglaucoma medication occurred in both groups (p < 0.05); these results were comparable in the two groups (p > 0.05). The mean IOP reduction was 34% in the eyes that underwent viscocanalostomy alone and 38% in the eyes that underwent phacoviscocanalostomy. The success rates of the two procedures were also comparable (p > 0.05). In the combined-surgery group the best-corrected visual acuity improved by 2 or more lines in 18 eyes (90%). Intraoperative complications did not occur in either group. INTERPRETATION: Combining phacoemulsification with viscocanalostomy did not have a negative effect on the IOP control achieved by viscocanalostomy alone and did not increase the complication rate. Phacoviscocanalostomy was safe and effective in the surgical management of eyes with both glaucoma and cataract. PMID- 14740801 TI - Earlier detection of amblyopia and strabismus by ophthalmologic screening card attached to the vaccination card. AB - BACKGROUND: In 1983, in Rijeka, Croatia, an ophthalmologic screening card was introduced for the detection and prevention of ophthalmologic diseases, including amblyopia and strabismus, in children. The card was attached to the vaccination card. The main goal of this study was to investigate whether this model of screening decreased the age at which children were first admitted to our Department of Strabismus and Pediatric Ophthalmology. METHODS: We randomly selected 100 of the 225 children who underwent examination for the first time in the Department of Pediatric Ophthalmology and Strabismus of a clinical hospital centre in Rijeka in 1980, and 100 of the 412 examined for the first time in 1990. Only children aged 5 years or less who were born in the maternity hospital in Rijeka were included in the study. We reviewed the records for these groups, noting the sex, age, diagnosis, refraction and visual acuity. RESULTS: The average age of the children examined in 1980 was 4.4 (standard deviation 1.4) years, compared with 2.5 (standard deviation 1.2) years in 1990, a statistically significant difference (p < 0.01, Student's t-test). In 1980, 17% of the children were under 3 years, compared with 80% in 1990 (p << 0.01). INTERPRETATION: The ophthalmologic screening card contributed to reducing the age at which strabismus and amblyopia are detected. This method of detection has been applied to the entire child population, and detection is performed continuously. PMID- 14740802 TI - Probable autoimmune retinopathy in a patient with multiple sclerosis. PMID- 14740803 TI - Intravitreal methotrexate injections for intraocular involvement in primary central nervous system lymphoma. PMID- 14740804 TI - Acute macular neuroretinopathy and associated disc swelling and blind spot enlargement. PMID- 14740805 TI - A rare agent of chalazion: demodectic mites. PMID- 14740806 TI - Vitreous hemorrhage as the presenting sign of antiphospholipid syndrome. PMID- 14740807 TI - Epiretinal membrane development during interferon treatment. PMID- 14740808 TI - Intraocular leukemia as the primary manifestation of relapsing acute myelogenous leukemia. PMID- 14740809 TI - Anatomy for the acupuncturist--facts & fiction. 3: Upper & lower extremity. AB - Anatomy knowledge, and the skill to apply it, is arguably the most important facet of safe and competent acupuncture practice. The authors believe that an acupuncturist should always know where the tip of their needle lies with respect to the relevant anatomy so that vital structures can be avoided and so that the intended target for stimulation can be reached. This article describes the anatomy of the upper limb and shoulder girdle, and lower limb and pelvis, relevant to safe needling practice. PMID- 14740810 TI - Comparison of transient heart rate reduction associated with acupuncture stimulation in supine and sitting subjects. AB - We investigated the difference in transient heart rate reduction associated with brief acupuncture in 20 healthy subjects at rest in a supine and in a sitting position. After the subjects had been at rest for about 20 minutes, acupuncture needling using the sparrow-pecking method, in which the needle is moved vertically lifting and thrusting, was performed for one minute at the Shousanli point on the right forearm (LI10). The procedure was carried out with the subjects in a supine position and in a sitting position. The position for stimulation of each subject, either supine or sitting, was selected at random, and on different days. The results showed that the average heart rate reduction associated with stimulation in supine subjects was 3.6 +/- 0.19 (mean +/- standard error (SE)) beats per minute (bpm), while that for sitting subjects was about 7.0 +/- 1.07 (mean +/- SE) bpm, indicating that stimulation reduces heart rate to a greater degree in subjects who are sitting (p<0.05, Mann-Whitney test). These results would be consistent with a mechanism involving reduced sympathetic drive to the heart, as sympathetic nerve activity has more influence on the heart rate in the sitting than in the supine position. PMID- 14740811 TI - A case series of temporomandibular disorders treated with acupuncture, occlusal splint and point injection therapy. AB - A treatment regime combining acupuncture, occlusal splint and point injection therapy for temporomandibular disorders (TMD) is presented. There were 89 consecutive patients treated by the regime in this case series but four patients dropped out after two to three visits. Data and treatment results of the remaining 85 patients who had treatment completed were analysed. It was found that 73 (85%) of patients with TMD had symptoms relieved within six visits under this regime. Complications were rare and minor. Acupuncture treatment, in combination with splint therapy and point injection therapy, appears to be effective for managing TMD. However, further research, using randomised controlled trials should be conducted to ascertain its effectiveness over other treatment modalities. PMID- 14740812 TI - Acupuncture for chronic venous ulceration. AB - Acupuncture was used to treat a 69-year-old man for bilateral ankle pain related to his rheumatoid arthritis. This led to a dramatic improvement in one of his chronic venous leg ulcers. There is very little recent literature on such cases, where acupuncture may be a useful additional treatment. PMID- 14740813 TI - Anosmia treated with acupuncture. AB - This is a report detailing the successful treatment of a case of anosmia with acupuncture. The patient was managed conventionally for two years with no sign of improvement. She regained the sense of smell following one session of acupuncture. Such patients should be investigated for any detectable organic cause prior to treatment with acupuncture. PMID- 14740814 TI - Acupuncture for the treatment of sweating associated with malignancy. AB - Acupuncture was used to treat a 60-year old woman with unexplained sweating associated with inoperable lung cancer that prevented her from sharing a bed with her husband. Other measures failed to improve her sweating, but she responded well to a course of acupuncture allowing her to continue sharing the marital bed. PMID- 14740815 TI - Rheological properties of concentrated skim milk: importance of soluble minerals in the changes in viscosity during storage. AB - Properties of condensed milks prior to spray drying dictate to a large extent the functionality of the resulting milk powder. Rheological properties of concentrated skim milk, with total solids content of 45% but different mineral content, were studied as a function of shear rate and storage time at 50 degrees C. These milks are proposed as a model to study the effects of minerals on rheology and age gellation of condensed milk prior to drying. During storage of the concentrated milk, the apparent viscosity, particularly after 4 h, increased markedly at all shear rates studied. The yield stress also increased steeply after 4 h of storage at 50 degrees C. The changes in apparent viscosity of concentrated milk stored for up to 4 h were largely reversible under high shear, but irreversible in samples stored for longer time. The appearance of yield stress suggested the presence of reversible flocculation arising from weak attraction between casein micelles, with a transition from reversible to irreversible aggregation during storage. Particle size analysis confirmed irreversible aggregation and fusion of casein micelles during storage. Gradual reduction of mineral content of concentrated milks resulted in a marked decrease in the apparent viscosity and casein micelle aggregation during storage, while addition of minerals to milk had the opposite effect. The results demonstrated that the soluble mineral content is very important in controlling the storage induced changes in the rheology of concentrated milks. PMID- 14740816 TI - Milk pH as a function of CO2 concentration, temperature, and pressure in a heat exchanger. AB - Raw skim milk, with or without added CO2, was heated, held, and cooled in a small pilot-scale tubular heat exchanger (372 ml/min). The experiment was replicated twice, and, for each replication, milk was first carbonated at 0 to 1 degree C to contain 0 (control), 600, 1200, 1800, and 2400 ppm added CO2 using a continuous carbonation unit. After storage at 0 to 1 degree C, portions of milk at each CO2 concentration were heated to 40, 56, 72, and 80 degrees C, held at the desired temperature for 30 s (except 80 degrees C, holding 20 s) and cooled to 0 to 1 degree C. At each temperature, five pressures were applied: 69, 138, 207, 276, and 345 kPa. Pressure was controlled with a needle valve at the heat exchanger exit. Both the pressure gauge and pH probe were inline at the end of the holding section. Milk pH during heating depended on CO2 concentration, temperature, and pressure. During heating of milk without added CO2, pH decreased linearly as a function of increasing temperature but was independent of pressure. In general, the pH of milk with added CO2 decreased with increasing CO2 concentration and pressure. For milk with added CO2, at a fixed CO2 concentration, the effect of pressure on pH decrease was greater at a higher temperature. At a fixed temperature, the effect of pressure on pH decrease was greater for milk with a higher CO2 concentration. Thermal death of bacteria during pasteurization of milk without added CO2 is probably due not only to temperature but also to the decrease in pH that occurs during the process. Increasing milk CO2 concentration and pressure decreases the milk pH even further during heating and may further enhance the microbial killing power of pasteurization. PMID- 14740817 TI - Proteolysis on Reggianito Argentino cheeses manufactured with natural whey cultures and selected strains of Lactobacillus helveticus. AB - Reggianito Argentino cheese is traditionally manufactured with whey starter cultures that provide typical and intense flavor but can cause poor quality standardization. In this study, the influence of natural and selected starters on Reggianito Argentino cheese proteolysis was investigated. Cheeses were manufactured with three strains of Lactobacillus helveticus (SF133, SF138 and SF209) cultured individually in sterile whey and used as single or mixed starters. Control cheeses were made with natural whey starter culture. Cheeses were analyzed to determine gross composition, as well as total thermophilic lactic flora. Proteolysis was assessed by N fractions, electrophoresis and liquid chromatography. Gross composition of the cheeses did not significantly differ, while viable starter cell counts were lower for cheeses made with strain SF209 alone or combined with other strains. Soluble N at pH 4.6 was the same for cheeses made with natural or selected starters, but soluble N in 12% trichloroacetic acid and 2.5% phosphotungstic acid was significantly higher in cheeses made with starters containing strain SF209. Nitrogen fractions results indicated that natural whey starter cultures could be replaced by several starters composed of the selected strains without significant changes to proteolysis patterns. Starter cultures prepared only with SF209 or with the three selected L. helveticus strains produced cheese products with significantly more proteolysis than control cheeses. Chromatographic profiles analyzed by principal components showed that three main peaks on chromatograms, presumptively identified as Tyr, Phe, and Trp, explained most of variability. Principal component scores indicated that cheese samples were grouped by ripening time, which was confirmed by linear discriminant analysis. On the contrary, samples did not cluster by Lactobacillus strain or type of starter. PMID- 14740818 TI - Use of dry milk protein concentrate in pizza cheese manufactured by culture or direct acidification. AB - Milk protein concentrate (MPC) contains high concentrations of casein and calcium and low concentrations of lactose. Enrichment of cheese milk with MPC should, therefore, enhance yields and improve quality. The objectives of this study were: 1) to compare pizza cheese made by culture acidification using standardized whole milk (WM) plus skim milk (SM) versus WM plus MPC; and 2) compare cheese made using WM + MPC by culture acidification to that made by direct acidification. The experimental design is as follows: vat 1 = WM + SM + culture (commercial thermophilic lactic acid bacteria), vat 2 = WM + MPC + culture, and vat 3 = WM + MPC + direct acid (2% citric acid). Each cheese milk was standardized to a protein-to-fat ratio of approximately 1.4. The experiment was repeated three times. Yield and composition of cheeses were determined by standard methods, whereas the proteolysis was assessed by urea polyacrylamide gel electrophoresis (PAGE) and water-soluble N contents. Meltability of the cheeses was determined during 1 mo of storage, in addition to pizza making. The addition of MPC improved the yields from 10.34 +/- 0.57% in vat 1 cheese to 14.50 +/- 0.84% and 16.65 +/- 2.23%, respectively, in vats 2 and 3 and cheeses. The percentage of fat and protein recoveries showed insignificant differences between the treatments, but TS recoveries were in the order, vat 2 > vat 3 > vat 1. Most of the compositional parameters were significantly affected by the different treatments. Vat 2 cheese had the highest calcium and lowest lactose contencentrations. Vat 3 cheese had the best meltability. Vat 1 cheese initially had better meltability than vat 2 cheese; however, the difference became insignificant after 28 d of storage at 4 degrees C. Vat 3 cheese had the softest texture and produced large-sized blisters when baked on pizza. The lowest and highest levels of proteolysis were found in vats 2 and 3 cheeses, respectively. The study demonstrates the use of MPC in pizza cheese manufacture with improved yield both by culture acidification as well as direct acidification. PMID- 14740819 TI - Microbiological quality of raw goat's and ewe's bulk-tank milk in Switzerland. AB - A total of 407 samples of bulk-tank milk (344 of goat's milk and 63 of ewe's milk) collected from 403 different farms throughout Switzerland, was examined. The number of farms investigated in this study represents 8% of the country's dairy-goat and 15% of its dairy-sheep farms. Standard plate counts and Enterobacteriaceae counts were performed on each sample. Furthermore, the prevalence of Staphylococcus aureus, Campylobacter spp., Shiga toxin-producing Escherichia coli, Salmonella spp., and Mycobacterium avium ssp. paratuberculosis was studied. The median standard plate count for bulk-tank milk from small ruminants was 4.70 log cfu/ml (4.69 log cfu/ml for goat's milk and 4.78 log cfu/ml for ewe's milk), with a minimum of 2.00 log cfu/ml and a maximum of 8.64 log cfu/ml. Enterobacteriaceae were detected in 212 (61.6%) goat's milk and 45 (71.4%) ewe's milk samples, whereas S. aureus was detected in 109 (31.7%) samples of goat's milk and 21 (33.3%) samples of ewe's milk. Campylobacter spp. and Salmonella spp. were not isolated from any of the samples. However, 16.3% of the goat's milk and 12.7% of the ewe's milk samples were polymerase chain reaction (PCR)-positive for Shiga toxin-producing E. coli. Seventy-nine (23.0%) goat's tank-milk and 15 (23.8%) ewe's tank-milk samples were PCR-positive for insertion sequence 900, providing presumptive evidence for the presence of M. avium ssp. paratuberculosis. These results form the basis for determining the microbiological quality standards for goat's and ewe's milk. Moreover, the data presented form part of the risk assessment program for raw milk from small ruminants in Switzerland. PMID- 14740820 TI - Effects of different levels of gum arabic, low methylated pectin and xylan on in vitro digestibility of beta-lactoglobulin. AB - Plant hydrocolloids used in the food industry to improve texture and stability of food, such as dairy products, can reduce protein digestibility and, consequently, modify the bioavailability of amino acids. We studied the in vitro hydrolysis at 37 degrees C of beta-lactoglobulin (beta-lg) in mixed dispersions containing either gum arabic or low-methylated pectin or xylan at levels of 0, 1, 10, 20, 30, and 50% weight. Proteolysis used either pepsin alone by progressive reduction of pH during proteolysis or pepsin followed by trypsin and chymotrypsin in two different dialysis bags with a molecular weight (MW) cutoff of 1000 or 8000 Da. Results showed that beta-lg was almost resistant to pepsin digestion and that the three plant hydrocolloids inhibited significantly beta-lg digestibility as determined using dialysis bag with a 1000-Da MW cutoff. Among the three polysaccharides used, xylan showed a digestibility decrease greater than that obtained with gum arabic and low-methylated pectin. On the other hand, no significant effect of polysaccharides on the in vitro beta-lg digestibility was detected using the dialysis bag with an 8000 Da MW cutoff. This mainly suggests that peptides with MW in the range 1000 to 8000 Da may interact with polysaccharides more than peptides and proteins with a greater molecular weight to decrease the protein digestibility, and that the nature of the polysaccharides plays a role in the interaction. PMID- 14740821 TI - Studies on physicochemical and functional properties of commercial sweet whey powders. AB - The objective of this study was to characterize variation and interrelatedness among primary functional and compositional parameters of commercially available sweet whey powders. Samples representing different plants/processes and cheese types were assayed for foaming capacity, foam stability, pH, protein content, soluble protein, turbidity, color, particle size distribution, lipid, and moisture. Data were analyzed using principal component analysis. Foaming capacity and stability varied from 10 to 220% and 0.1 to 14 min, respectively. Protein content and solubility ranged from 8.5 to 17.6% and 3.7 to 14.1%, respectively. Lipid content of sweet whey powder varied from 0.03 to 2.00%. The two main functional properties, foaming and protein solubility, did not show significant correlation with each other. Foaming properties showed a positive correlation to particle size and L* or lightness value, and negative correlation to lipid content. Protein solubility showed positive correlation with protein content and negative correlation with turbidity of the sample. Foaming behavior, protein, and particle size attributes were the main variables responsible for grouping of samples. Sweet whey powders from the same dairy plants were grouped together. The direct or indirect significance of these relationships to processing is detailed in this study. PMID- 14740822 TI - Dependence of the interfacial behavior of beta-casein on phosphoserine residues. AB - The role of the phosphoserine residues on the dynamical and structural properties of beta-casein was studied by molecular dynamics of the protein in water/lipid interfacial regions. The initial protein structure adopted in the modeling was that proposed for bovine beta-casein A2, where the five phosphoserine residues, originally present in its primary structure, were partially or totally substituted by serine residues. The simulations revealed a dependence of the interfacial behavior of beta-casein on the phosphorylation grade. When only partially dephosphorylated, the protein showed a similar behavior as that observed for the original beta-casein reported in previous work. During dynamics, the protein migrated from the aqueous environment towards the lipid medium, and remained attached to the interface separating both media. Quite different was the dynamics of the totally dephosphorylated beta-casein, that did not perceive the interface and immersed incessantly into lipid medium. The results suggest that the phosphoserine residues appear to be, in fact, intrinsically related to the mechanisms of beta-casein emulsion stabilization. PMID- 14740823 TI - Ventilation effects on air quality and on the yield and quality of ewe milk in winter. AB - The effects of ventilation on air quality, and on the welfare and production performance of dairy ewes were assessed in a 6-wk trial conducted during the winter of 2002. Thirty-six midlactation Comisana ewes were divided into three groups of 12, which were randomly balanced for parity, time of lambing, and number of lambs suckled. Treatments were low (LOV), moderate (MOV), and programmed ventilation regimen (PROV). In LOV and MOV rooms, fans provided 10 ventilation cycles of 40 min each at a fan speed of 1 and 2 m/s, respectively. In the PROV room, the fan was programmed to maintain a 70% relative humidity. Mean ventilation rates were 23, 47, and 73 m3/h per ewe in LOV, MOV, and PROV rooms, respectively. Air concentrations of microorganisms and dust, and of gaseous pollutants were measured twice weekly. Cell-mediated immune response to phytohemagglutinin at d 1, 21 and 42, and humoral response to chicken egg albumin at d 11, 21, 30, and 40 were determined. At d 39, ewes were injected with 2 IU of porcine adreno-corticotropic-hormone/kg body weight(0.75), and subjected to blood sampling for evaluation of cortisol concentrations immediately before and 1, 2, and 4 h after adreno-corticotrophic-hormone injection. Milk yield was recorded daily. Individual milk samples were analyzed weekly for composition, renneting parameters, and somatic cell count. The LOV treatment resulted in higher air concentrations of NH3 and CO2 than the MOV and PROV treatments. Greater amounts of total and respirable dust were found in the PROV room than in the LOV and the MOV rooms. The LOV ewes had lower milk yield than the PROV ewes, lower milk casein content, and higher rate of clot formation than the MOV and PROV ewes. The ventilation regimen did not affect the immune and endocrine responses of the ewes. Results suggest that an intermittent ventilation regimen, providing a mean ventilation rate of 47 m3/h per ewe at a fan speed of 2 m/s, is required to sustain the yield and cheese-making ability of ewe milk during the winter season. PMID- 14740824 TI - Influence of vacuum level and overmilking on udder health and teat thickness changes in dairy ewes. AB - The effects of vacuum level and overmilking on udder health were studied in ewes. Vacuum levels of 36 and 42 kPa were assigned to two groups of 23 Manchega ewes in a crossover study design with two experimental periods of 5 wk for each. Moreover, for each ewe, one teat was overmilked 1.5 to 2 min at all milkings during these 10 wk. The milking machine used had a midlevel milkline and pulsation was fixed at 180 cycles per min and a pulsation ratio of 50:50. Bacterial exposure of all teats was increased by dipping them in a suspension of Staphylococcus simulans at eight milkings of each period. New intramammary infections (IMI) were not significantly affected by the vacuum level used (18 and 23% of ewes infected, at 36 and 42 kPa, respectively) or application of overmilking (9 and 11% of half udders infected without and with overmilking, respectively). Likewise, neither factor significantly affected the somatic cell count (SCC) of the milk. Teat thickness changes after milking varied significantly due to the presence of overmilking (-13.6 and -7.4%, in teats not overmilked and overmilked, respectively) but were not affected by vacuum level. At no time were any lesions or variations visibly noted in the teat walls or orifice. So, in this work we were unable to demonstrate that the vacuum and overmilking levels assayed, both used with a pulsation rate of 180 cycles/min, have an important effect on the state of udder health in the short term. Furthermore, it was also observed that, in absence of IMI, the two factors studied did not cause irritation of any kind in the gland that might influence the SCC of the milk. PMID- 14740825 TI - Effectiveness of an internal teat seal in the prevention of new intramammary infections during the dry and early-lactation periods in dairy cows when used with a dry cow intramammary antibiotic. AB - The objectives of this study were to determine the effect of infusion with an internal teat seal at dry off, when used as an adjunct to long-acting antibiotic infusion at dry off, on the risk for acquiring a new intramammary infection (IMI) during the dry period, prevalence of IMI and linear score (LS) after calving, and risk for experiencing a clinical mastitis event between dry off and 60 DIM. A total of 437 cows from 2 dairy herds, with no clinical mastitis and 4 functional quarters, were enrolled at dry off. Prior to the final milking, all quarters were sampled for bacteriological culture and SCC analysis. After milking, all 4 quarters were infused with a commercially available long-acting dry cow antibiotic. Two contralateral quarters were then infused with an internal teat seal (Orbeseal, Pfizer Animal Health, New York). Following calving the teat seal was stripped out at first milking. Duplicate milk samples were collected between 1 to 3 DIM and again between 6 to 8 DIM for culture and SCC analysis. Quarters treated with Orbeseal had significantly lower prevalence of IMI at 1 to 3 DIM (tx = 22.8%, control = 29.1%), had significantly fewer quarters that acquired a new IMI between dry off and 1 to 3 DIM (tx = 20.2%, control = 25.4%), and had significantly fewer quarters affected by a clinical mastitis event between dry off and 60 DIM (tx = 5.9%, control = 8.0%). Multivariable analysis showed a significant effect of treatment, with treated quarters being 30% less likely to develop a new IMI between dry off and 1 to 3 DIM, 31% less likely to have an IMI present at 1 to 3 DIM, 33% less likely to experience a clinical mastitis event between dry off and 60 DIM, and having significantly lower linear score measures at 1 to 3 DIM and 6 to 8 DIM, compared with control quarters. PMID- 14740826 TI - Prophylactic effects of two selective dry cow strategies accounting for interdependence of quarter. AB - Infusion of a long-acting antibiotic preparation at drying off in dairy cows as a prophylactic therapy is usually recommended for all quarters where it is in use. Studying the effectiveness of such treatment using quarter as the unit of analysis assumes that each quarter within a cow has a risk of being infected independent of the other quarters of the cow. Failure to account for interdependence of quarters within a cow may lead to inaccurate variance estimates and errors in assessing treatment effects. Data from two trials assessing different dry-cow strategies were examined for interdependence of infection between quarters. Logistic regression with a variance inflation factor or a multilevel analysis was used to assess the effect of antibiotic and internal teat-sealant dry cow strategies. Parity and infection status at drying off were covariates in the analysis. Interdependence of the risk of quarter infections within control-group cows was demonstrated in both dry-cow antibiotic and teat seal trials. However, cows that received either of these treatments did not demonstrate interdependence. Treated quarters in both trials were 3.0 times less likely to acquire a new infection at calving compared with the untreated controls. Quarters in cows of parity 3 or greater were also at an increased risk in the antibiotic treatment trial. In both trials, quarters with either Corynebacterium spp. or coagulase-negative staphylococci infections at drying off had an increased risk of a new intramammary infection at calving. This study has demonstrated the beneficial and comparable effects of antibiotic and teat seal dry cow strategies; both decreased the risk of intramammary infection at calving. The application of dry-cow strategies at the cow level and not the quarter level is also supported. PMID- 14740827 TI - Plasma hormones and expression of growth hormone receptor and insulin-like growth factor-I mRNA in hepatic tissue of periparturient dairy cows. AB - Growth hormone plays a central role in the change in nutrient metabolism that occurs during the initiation of lactation. The actions of growth hormone are mediated by the growth hormone receptor (GHR) whose mRNA is present in three alternatively spliced forms (GHR 1A, 1B, and 1C). Liver-specific GHR 1A mRNA is transiently decreased around parturition, but the exact timing of the decline is not known. Our objective was to generate a daily profile for total GHR (GHRtot; all GHR transcripts), GHR 1A, and IGF-I mRNA expression in liver of periparturient Holstein cows and evaluate these daily mRNA profiles relative to daily profiles for periparturient hormones and metabolites. Liver biopsies and blood samples (n = 139) were collected from 65 Holstein cows at the University of Missouri Dairy Farm. At least two cows were sampled on each day from 14 d before to 14 d after parturition. Total cellular RNA was isolated and reverse transcribed to cDNA. Target cDNA were measured by quantitative real-time polymerase chain reaction. Plasma was assayed for progesterone, estradiol, insulin, growth hormone, IGF-I, glucose, and nonesterified fatty acids. The GHR 1A mRNA declined 2 d before parturition, was lowest 3 to 4 d after parturition, and then increased. The IGF-I mRNA declined 1 d after parturition, was lowest 2 to 5 d after parturition and then increased. Total GHR mRNA was not affected by day. The decrease in GHR 1A mRNA was associated with a decrease in progesterone and an increase in estradiol shortly before parturition. A detailed profile of GHR 1A, IGF-I, and GHRtot mRNA expression during the periparturient period was provided. The decreases in GHR 1A and IGF-I during the transition period occurred immediately before (GHR 1A) or shortly after (IGF-I) parturition. Rapid changes in placental and ovarian steroids before parturition were coincident with changes in GHR 1A mRNA. PMID- 14740828 TI - In vitro antimicrobial susceptibility of Escherichia coli isolates from clinical bovine mastitis in Finland and Israel. AB - Minimal inhibition concentration (MIC) values of 100 Finnish and 100 Israeli Escherichia coli isolated from clinical bovine mastitis were determined for ampicillin, cephalexin, ceftazidime, dihydrostreptomycin, gentamicin, tetracycline, trimethoprim-sulfadiazine, and ciprofloxacin by an agar dilution method. The in vitro antimicrobial susceptibility of the E. coli isolates was high; only 27% showed resistance to one or more tested antimicrobial agents. Fifteen percent of the Israeli isolates and 14% of the Finnish isolates were resistant to tetracycline, 3 and 16% to cephalexin, 10 and 7% to ampicillin, 13 and 9% to dihydrostreptomycin, and 4 and 2% to trimethoprim-sulfadiazine. No gentamicin-, ceftazidime-, or ciprofloxacin-resistant isolates were detected. Eleven percent of all the isolates were resistant to two or more antimicrobial agents. Tetracycline was most often associated with multiresistant patterns. Most of the multiresistant isolates had very high MIC values, whereas most of those that were resistant to only one tested antibiotic had MIC values close to the susceptibility breakpoint. Antimicrobial resistance appeared to pose no problem in E. coli isolated from mastitic milk of both countries. This is probably due to the controlled use of antimicrobial agents in the treatment of dairy herds. Some differences were present in the resistance patterns, which may reflect the different use of antimicrobial agents in these two countries. PMID- 14740829 TI - Growth hormone (GH) binding and expression of GH receptor 1A mRNA in hepatic tissue of periparturient dairy cows. AB - Growth hormone (GH) plays a role in metabolic adaptations that occur during lactogenesis. Liver GH receptor transcript (GHR 1A) is transiently decreased near parturition and may reduce GH-dependent signaling leading to low blood insulin like growth factor I (IGF-I) concentrations in periparturient dairy cattle. We hypothesized that the decrease in GHR 1A mRNA at parturition was associated with decreased GH binding (i.e., GHR protein concentration) in liver. Blood and liver biopsy samples were collected from 12 Holstein cows on d -12 +/- 1, 3, and 17 relative to parturition. Total cellular RNA was isolated from a sub-sample of liver. Quantitative real-time polymerase chain reactions were used to measure GHR 1A, total GHR, IGF-I, and cyclophilin mRNA. Microsomal membranes were isolated from the remaining liver tissue and assayed for 125I-bGH binding. Plasma was assayed for GH and IGF-I concentrations. Liver GHR 1A mRNA, specific 125I-bGH binding to liver membranes, liver IGF-I mRNA, and plasma IGF-I concentrations were lower on d 3 relative to d -12. The GHR 1A mRNA, 125I-bGH binding, and plasma GH concentrations increased on d 17 but liver IGF-I mRNA and plasma IGF-I concentrations did not change between d 3 and 17. Total GHR mRNA and cyclophilin mRNA amounts were similar on d -12, 3, and 17. Across all days, 125I-bGH specific binding in liver was highly correlated with liver GHR 1A mRNA (R2 = 0.68) but not with total GHR mRNA. Saturation binding analysis showed that GHR concentration (Bmax) in liver on d 3 had decreased to only 5% of the amount on d -12. We conclude that decreased GHR 1A mRNA leads to decreased GHR protein concentration in liver. Reduced GHR in liver likely contributes to a decrease in liver IGF-I production and reduced concentrations of IGF-I in blood of periparturient cows. PMID- 14740830 TI - Fertility of dairy cows after resynchronization of ovulation at three intervals following first timed insemination. AB - Lactating Holstein cows (n = 711) on a commercial dairy farm in Wisconsin received a hormonal synchronization protocol to initiate first timed artificial insemination (TAI) on the following postpartum schedule: two injections of 25 mg PGF2alpha at 32 +/- 3 d and 46 +/- 3 d (Presynch); 100 microg GnRH at 60 +/- 3 d; 25 mg PGF2alpha at 67 +/- 3 d; and 100 microg GnRH + TAI at 69 +/- 3 d (Ovsynch). At first TAI, cows were randomly assigned to initiate the first GnRH injection of a hormonal protocol for resynchronization of ovulation (Resynch; 100 microg GnRH, d 0, 25 mg PGF2alpha, d 7, 100 microg GnRH + TAI, d 9) at 19 (D19), 26 (D26), or 33 d (D33) after first TAI to set up a second TAI service for cows failing to conceive to Ovsynch. Overall pregnancy rate per artificial insemination (PR/AI) to Ovsynch assessed 68 d after TAI was 31% and did not differ among treatment groups. For Resynch, PR/AI was assessed 26 d after TAI for D19 and D26 cows and 33 d after TAI for D33 cows. Overall PR/AI to Resynch was 32%. However, the PR/AI for D26 (34%) and D33 (38%) cows to Resynch was greater than for D19 cows (23%). Cows with a CL at the PGF2alpha injection (D19 cows) or at the first GnRH injection (D26 + D33 cows) of Resynch exhibited greater PR/AI to Resynch compared with cows without a CL. Survival analysis (failure time) of cows in the D26 and D33 treatment groups across the first three TAI services did not differ statistically. Although administration of GnRH to pregnant cows 19 d after first TAI service did not appear to induce iatrogenic embryonic loss, initiation of Resynch 19 d after first TAI service resulted in a lower PR/AI compared with initiation of Resynch 26 or 33 d after first TAI service. PMID- 14740831 TI - Short communication: Teat skin pH. AB - Changes in teat skin surface pH were studied over 12 wk in 99 lactating Holstein cows. Half the udder of each cow routinely received postmilking disinfection, and the other half served as control. Measures of pH were made on all teats at weekly intervals. Teat skin pH was affected by treatment but not week. Mean teat skin pH measures were: 7.18 (+/- 0.64) and 7.53 (+/- 0.46) for treatment and control teats. In study II, pH teat skin measures were made hourly on 16 cows, starting 2 h before milking, immediately before a milking, immediately after a milking, and for 2 h postmilking. Teat skin pH was significantly lower for treatment teats and was lower for all teats postmilking. PMID- 14740832 TI - Effects of biotin supplementation on performance and claw lesions on a commercial dairy farm. AB - A controlled 14-mo field trial was conducted to evaluate the effect of biotin supplementation on hoof lesions, milk production, and reproductive performance in a commercial dairy herd. One hundred seventy cows were studied and supplemented with either 0 or 20 mg/d of biotin by computer feeder. All were housed in the same free-stall facility with the same environment, base diet, and management. The feet of 99 cows were trimmed three times at 6-mo intervals, and hoof health was evaluated. Milk production and fertility data were captured monthly by the Dairy Herd Improvement Association. At the final hoof trimming, sole hemorrhages were significantly higher in control (50%) vs. biotin-supplemented animals (24%). The incidents of cows affected with double soles, hoof wall grooves, and heel horn erosion did not differ between control and biotin-supplemented animals. Biotin supplementation of trimmed cows resulted in 878 kg more milk than control cows when compared with previous lactation yield (n = 46 biotin supplemented, n = 48 control cows). At the end of the study, for both trimmed and untrimmed animals, biotin supplemented cows (n = 81) produced 481 kg more milk and 25 kg more fat than the controls (n = 81). There was no interaction between biotin supplementation and hoof trimming on milk production. There were variations in the response of fertility to biotin between age groups. First lactation heifers fed supplemental biotin had significantly fewer days from calving to conception and required fewer inseminations per pregnancy than controls of the same parity. PMID- 14740833 TI - Antibiotic resistance in gut bacteria from dairy calves: a dose response to the level of antibiotics fed in milk. AB - Dairy calves are commonly fed milk from cows treated with antibiotics. The concentration of beta-lactam antibiotic residues found in milk from treated cows was used to determine the range of concentrations of penicillin used in a dose regulated experiment. Thirty-one Holstein calves were randomly assigned to milk with penicillin G added at concentrations of 0, 6.25, 12.5, 25, and 50 microl/kg. Fecal swabs were taken from each calf twice weekly. Resistance to penicillin was tested by measuring the zone of inhibition in bacterial growth around a disk impregnated with the antibiotic. Inhibition was greatest for bacteria from calves fed milk with no penicillin (2.89 +/- 0.14 mm), and declined as the penicillin dose provided in the milk increased (to a low of 0.70 +/- 0.10 for the 50 microl/kg treatment group). In conclusion, resistance of gut bacteria to antibiotics increases with increasing concentrations of penicillin in the milk fed to dairy calves. PMID- 14740834 TI - Effect of lasalocid on growth, blood gases, and nutrient utilization in dairy goats fed a high forage, low protein diet. AB - This study investigated the effect of lasalocid on weight gain, blood gases, nutrient digestibility, and nitrogen utilization in growing dairy goats. In a growth experiment, 24 crossbred dairy does were assigned to diets without or with lasalocid (approximately 30 mg per head per day) for 12 wk. Goats were group fed bermudagrass hay for free choice plus concentrate at a fixed level (approximately 0.264 kg DM per head per d). Goats grew faster when fed lasalocid than those fed the control diet. Jugular blood partial pressure of O2 was lower when goats were supplemented with lasalocid. In contrast, goats fed lasalocid tended to have a higher partial pressure of CO2 than control goats. Percent O2 saturation tended to increase, and concentrations of total CO2 tended to decrease in goats fed lasalocid. Serum glucose and urea N did not differ between treatments. In a metabolism experiment, 8 castrated male goats were fed hay and concentrate identical to those of the growth experiment to determine whole tract nutrient digestibility and N utilization. Lasalocid did not affect feed intake or nutrient digestion coefficients. Daily urinary N output was reduced by lasalocid supplementation. In spite of this, N retention was not significantly affected. With use of ionophore, the proportion of urinary N relative to digested N tended to decrease, and the retained N as a proportion of digested N tended to increase. PMID- 14740835 TI - The effect of steam-flaked or dry ground corn and supplemental phytic acid on phosphorus partitioning and ruminal phytase activity in lactating cows. AB - The effect of starch source and phytic acid (PA) supplementation on phosphorus (P) partitioning and ruminal phytase activity was evaluated in eight midlactation cows (four ruminally cannulated). Cows were randomly assigned to treatments in replicated 4 x 4 Latin squares with four 18-d periods. Diets included dry ground corn (DG) or steam-flaked corn (SF), with no supplemental P (L; 0.33% P) or supplemental purified PA (0.44% P) to provide additional P from a nonmineral source. Total collection of milk, urine, and feces was conducted on d 16 to 18 of each period. Ruminal fluid was sampled and ruminal pH measured every 8 h on d 17 and 18. Milk yield was unaffected by starch source, despite lower DMI by cows fed SF. Cows fed SF had increased DM digestibility compared with those fed DG, and tended to have higher efficiency of milk yield (1.40 vs. 1.35 kg of milk/kg of DMI). Intake and fecal excretion of P was lower in cows fed SF than in cows fed DG. In cows fed SF, milk P as a percentage of P intake increased compared with cows fed DG. Ruminal pH was unaffected by diet, but milk fat content was lower for cows fed SF. Milk yield, DMI, and feed to milk ratio were not affected by supplementation with PA. Although cows fed PA had increased P intake compared with cows fed low P diet, increased P excretion resulted in no differences in apparent P digestibility. Phosphorus balance tended to be higher in cows fed PA, but milk P as a percentage of intake was reduced. The interaction of starch source and PA affected ruminal phytase activity. Altering starch source to improve efficiency of milk yield in lactating dairy cows may help reduce P losses from dairy farms. PMID- 14740836 TI - Influence of dietary nonfiber carbohydrate concentration and supplementation of sucrose on lactation performance of cows fed fescue silage. AB - There is interest in knowing if the source of nonfibrous carbohydrates (NFC) influences milk production and composition. Our objective was to determine the effects of source (starch or sugar) and level of NFC in the diet on these parameters. A 4 x 4 Latin square replicated five times using early-lactation (56 +/- 9 DIM) Holstein cows was used; cows were offered one of two levels of NFC and either no added sucrose or sucrose substituting for 10% of the corn. Diets were balanced to meet National Research Council requirements for total protein, energy, and minerals. Tall fescue silage was included at one of two levels (0.95 or 1.25% of BW as forage NDF), resulting in diets with 40 and 30% NFC. The remaining ingredients consisted of high-moisture corn, soybean meal, SoyPlus, minerals, and vitamins. Megalac (0.45 kg) was used in the low NFC diets. High NFC diets were lower (P < 0.01) in neutral detergent fiber (NDF; 31.5%) and crude protein (CP; 19.6%) than the low NFC diet (35.8% NDF and 21.0% CP). Sucrose containing diets were somewhat lower (P < 0.01) in NDF (33.1%) than the no sucrose added diets (34.3%), but diets did not differ in CP%. Cows offered the high NFC level produced more milk (39.6 kg/d; P < 0.05) than those offered the low level (38.3 kg/d), primarily due to higher dry matter intake (P < 0.05). Cows consuming the high NFC diet also had lower (P < 0.05) milk fat (3.25%) and milk urea nitrogen (MUN; 13.7 mg/dl), and higher (P < 0.05) milk protein (2.58%) and milk lactose (4.81%) concentrations than cows offered the low NFC level (3.46% milk fat, 17.5 mg/dl MUN, 2.51% milk protein, and 4.74% milk lactose). Fat yield was higher (P < 0.05) for cows fed low NFC diets than cows fed high NFC diets, whereas protein and fat yield were lower (P < 0.05) for cows fed low NFC diets than those fed high NFC diets. The NFC source did not influence dry matter intake or milk production or milk component yield (P > 0.05). Milk lactose (4.79%) and MUN (16.0 mg/dl) concentrations were higher (P < 0.05) for cows offered sucrose as a portion of the NFC compared with those not offered sucrose (4.76% milk lactose and 15.2 mg/dl MUN). Results suggest that cows fed sucrose may utilize diet nitrogen less efficiently than those not fed sucrose, when sucrose is replacing a portion of the high-moisture corn in the diet. PMID- 14740837 TI - Influence of nutrition level on digestibility in high yielding cows and effects on energy evaluation systems. AB - The objective of the study was to determine the effect of nutrition level (NL, multiples of maintenance energy requirement) on the digestibility of nutrients for dairy cows regarding the energy supply of the animal. The digestion of nutrients and energy was investigated in two trials using lactating dairy cows. The NL varied from 2.7 to 5.0 using diets similar composition. In addition, sheep were given the same feed with a NL of 1.4. Digestibility of dry matter (DM) and all specific measures of dietary components declined significantly as NL increased. Digestibility of energy decreased by 4.1% for each increase in NL. The metabolizable energy, the ability to metabolize energy (metabolizable energy/gross energy), and the content of net energy for lactation (NE(L)) per kilogram of DM intake were calculated for NL from 1 to 6 on the basis of these relationships and as well as the gross energy, methane energy, and urine energy. Accordingly the NEL content declined by 0.11 MJ/kg of DM intake or 1.6% as the NL increased by one unit. It means that the NE(L) requirement above the maintenance requirement increased by 0.07 MJ of NE(L) per kilogram of fat-corrected milk, if the NL increased by one unit. PMID- 14740838 TI - Effect of linseed oil supplementation on ruminal digestion in dairy cows fed diets with different forage:concentrate ratios. AB - The effect of linseed oil (LSO) supplementation on total-tract and ruminal nutrient digestibility, N metabolism, and ruminal fluid characteristics was investigated in dairy cows fed diets containing different forage to concentrate ratios (F:C). The experimental design was a 4 x 4 Latin square with 2 x 2 factorial arrangement of treatments. Four lactating Holstein cows were fed a forage-rich diet without LSO (F; F:C = 65:35, dry matter basis), a forage-rich diet with LSO (FO; F:C = 65:32, 3% LSO), a concentrate-rich diet without LSO (C; F:C = 35:65), or a concentrate-rich diet with LSO (CO; F:C = 35:62, 3% LSO). Total-tract digestibility of DM and OM was greater with supplemental LSO. A tendency for greater total-tract digestibility of NDF and ADF also was observed in cows fed LSO. Ruminal digestibility of NDF or ADF decreased when CO was fed compared with C. In contrast, feeding FO increased NDF or ADF digestibility compared with F. Although ruminal starch digestion was nearly complete with all diets, digestibility was greater when cows were fed C or CO compared with F or FO. Bacterial N flow to the duodenum decreased when FO was fed compared with F. In contrast, feeding CO increased bacterial-N flow compared with C. Neither F:C nor LSO supplementation affected ruminal pH or total VFA concentration in ruminal fluid. However, molar proportion of propionate was greater with C or CO compared with F or FO and increased with LSO supplementation regardless of F:C. Molar proportion of n-butyrate decreased with LSO supplementation. Total protozoal numbers in ruminal fluid decreased markedly only when CO was fed. Overall, data show that feeding LSO had no negative effects on total-tract digestion in dairy cows but may decrease ruminal fiber digestibility when fed with high-concentrate diets. The widely spread idea that LSO decreases digestibility, arising from studies with sheep, did not seem to apply to lactating cows fed 3% LSO. PMID- 14740839 TI - Nitrogen supplementation of corn silages. 1. Effects on feed intake and milk production of dairy cows. AB - Feed intake and milk production responses to N supplementation of corn silage based diets were measured in three 3 x 3 Latin square experiments. In each experiment, 9 Holstein cows received total mixed rations (TMR), based on corn silage. In Exp. 1, midlactation cows were used to study effects of diets with different ratios of effective rumen-degradable protein (ERDP; g) to fermentable metabolizable energy (FME; MJ), providing a large deficiency (RL), a slight deficiency (RM), and a slight excess (RH) in relation to the target level of 11 g of ERDP/MJ of FME, respectively, for lactating cows. Diets were formulated to be isoenergetic, and to satisfy the metabolizable protein requirements. In Exp. 2, early-lactation cows were used to evaluate effects of different proportions of quickly and slowly rumen-degradable protein (RDP), achieved by replacing soybean meal with urea in the concentrates (0, 0.5, and 1% urea). Experiment 3 investigated effects of synchronizing the availability of FME and ERDP in the rumen. Midlactation cows received a diet containing, on a dry matter (DM) basis, 45% corn silage, 5% ryegrass hay, 35% energy-rich concentrate, and 15% protein rich concentrate (crude protein: 38% of DM; urea: 2% of DM). The protein-rich concentrate was fed either once (D1) or twice (D2) per day before the meal, or included in the TMR (DU). Treatment RL led to lower DM intake and milk yield, but higher milk production efficiency; there were no significant differences between treatments RM and RH. There were no significant treatment effects on DM intake, milk yield, or milk composition in Exp. 2. Manipulating rumen synchrony by altering the timing of feeding affected milk yields, with D1 cows producing significantly less than D2 and DU cows, which were similar. The amount of ERDP in the diet should be matched to the amount of fermentable energy available to maximize intake, milk yields, and the conversion of feed N into milk protein. However, this study showed only small benefits to altering the diurnal pattern of supply of RDP and FME, and only with extreme feeding strategies that would not be used in practice. Urine volume increased in response to increased or unbalanced protein supply. Analysis of the allantoin:creatinine ratio in spot samples of urine was not useful in identifying predicted differences in microbial protein yield from the rumen. PMID- 14740840 TI - Nitrogen supplementation of corn silages. 2. Assessing rumen function using fatty acid profiles of bovine milk. AB - The effects of N supplementation strategies on milk fatty acid profiles of dairy cows and their use as a noninvasive technique to diagnose rumen function, and to guide protein feeding decisions on-farm were evaluated in three experiments. Each experiment was designed according to three 3 x 3 Latin squares with 9 Holstein cows receiving total mixed rations based on corn silage. Experiment 1 was designed to study effects of diets with different ratios of effective rumen degradable protein (ERDP; g) to fermentable metabolizable energy (FME; j) providing, respectively, a large deficiency, a slight deficiency, and a slight excess in relation to the target level of 11 g of ERDP/MJ FME for lactating cows. Experiment 2 evaluated effects of different proportions of quickly and slowly rumen-degradable protein achieved by replacing soybean meal with urea in the concentrates (0, 0.5, and 1% urea for U0, U5, and U10, respectively). Experiment 3 investigated effects of synchronizing the availability of FME and ERDP in rumen by offering the protein-rich concentrate once or twice per day before the meal (corn silage, ryegrass hay, and energy-rich concentrate), or included in the total mixed ration. Milk fatty acid profiles were significantly affected by dietary N and carbohydrate supply. Principal component factor analysis provided a reasonable description of the data, clearly discriminating between fatty acids that are synthesized by different metabolic pathways. Several sources/pathways were distinguished: de novo synthesis in the mammary gland (short- and medium chain fatty acids), delta9-desaturase activity (monoenoic fatty acids), direct absorption from the blood stream (long-chain fatty acids), and de novo synthesis by the rumen microbial populations (odd-chain fatty acids). Discriminant canonical analysis showed that milk odd-chain fatty acids had a higher ability to discriminate between diets than even-chain fatty acids. The anteiso C15:0 increased in line with increasing sugar supply, and C17:0 appears to be a marker of protein deficiency. Additionally, iso C17:0 and anteiso C17:0 were associated with the NDF and CP contents of diets. The results suggests that milk odd-chain fatty acids have the potential to be used as a noninvasive technique to assess rumen function in terms of microbial populations, substrates and interactions. PMID- 14740841 TI - Effect of two levels of crude protein and methionine supplementation on performance of dairy cows. AB - Sixteen Holstein cows in midlactation were randomly assigned to treatments in a replicated 4 x 4 Latin square. Two levels of CP (16.1 vs. 18.8%) with or without supplemental methionine (0.07 g/100 g of DM) were tested in a 2 x 2 factorial arrangement of treatments. Dry matter intake, milk production, milk composition, and N excretion were determined. No interactions between CP level and methionine supplementation were observed. Milk production and dry matter intake were not different among treatments. Milk protein concentration increased from 3.17 to 3.26% with the addition of methionine and decreased from 3.24 to 3.17% with increased CP. No differences were observed among treatments in milk protein yield. Milk fat concentration was low across all diets, but was increased from 2.33% with 16.1% CP diets to 2.68% with 18.8% CP diets. No significant treatment effects were observed for SNF, lactose concentration in milk, or casein N as a fraction of skim milk N. Increased dietary CP increased milk urea N by 3.9 mg/dl. Methionine supplementation did not affect N excretion in urine or feces. The higher protein diets increased estimated urine volume by 2.9 L/d and increased N concentration by 1.7 percentage units in both urine and feces. Feeding higher protein increased milk urea and urine N excretion as expressed as a percentage of total N excreted (44 vs. 38% for 18.8 and 16.1% CP, respectively). Overall, feeding 16.1% CP produced milk and milk protein yields similar to feeding 18.8% CP, but reduced the N losses in urine and milk urea. PMID- 14740842 TI - Short communication: Effects of a monensin premix on milk fatty acid content during subacute ruminal acidosis in dairy cows. AB - The effects of a monensin premix on milk fatty acid content during grain-induced subacute ruminal acidosis (SARA) in Holstein cows receiving a total mixed ration was investigated. Six multiparous, rumen-fistulated Holstein cows were used in a two-treatment, two-period crossover design with 6-wk periods. Experimental treatments were either a monensin premix or a placebo premix. At the beginning of wk 4, SARA was induced in experimental cows for a 10-d period using a grain challenge model. The administration of a monensin premix elevated milk fat proportion of total short-chain saturated fatty acids (sum of C4 to C15). Milk fat proportions of conjugated linoleic acid isomers were unaffected. Linolenic acid (C18:3n3) proportion in milk fat of monensin-treated cows were lower when compared with placebo-treated cows during the SARA period. Results from this study indicate that dietary supplementation with monensin during SARA had little effect on milk fatty acid content. PMID- 14740843 TI - Use of principal component analysis to investigate the origin of heptadecenoic and conjugated linoleic acids in milk. AB - The aim of this paper was the application of principal component analysis (PCA) 1) to elucidate mutual metabolic relationships between milk fatty acids (FA) and 2) to illustrate the origin of milk FA, in particular C17:1 and cis-9,trans-11 conjugated linoleic acid. Data were combined from 3 experiments with lactating Holstein-Friesian cows offered diets based on grass or legume silage and concentrates. Loading plots of PCA based on milk FA concentrations showed 4 groups of milk FA, having similar precursors or metabolic pathways in the rumen and/or mammary gland: medium-chain saturated FA, de novo synthesized from acetate and beta-hydroxybutyrate; monoenoic milk FA, products of delta9-desaturase activity in the mammary gland; odd chain FA of rumen microbial origin and C18:0, n-6 C18:2, and n-3 C18:3 of dietary origin or the result of rumen biohydrogenation. Loading plots of PCA based on both milk and duodenal FA concentrations as well as on milk FA yields and duodenal FA flows further illustrated the importance of postabsorptive synthesis of the milk medium chain saturated and monoenoic FA and the direct absorption from the blood stream of odd chain FA, C18:0, n-6 C18:2, and n-3 C18:3. In all loading plots, milk oleic acid (C18:1) appeared intermediate between clusters of 18-carbon FA and monoenoic FA, illustrating its dual (dietary and endogenous production) origin. Milk C17:1 was suggested to be a desaturation product of C17:0, in common with other milk monoenoic FA. Finally, the PCA technique, based on milk FA patterns of one experiment, was applied to investigate factors determining cis-9,trans-11 conjugated linoleic acid concentrations in milk. Within the range of diets and cows studied here, we showed changes in cis-9,trans-11 conjugated linoleic acid to be mainly dependent on vaccenic acid supply and to a lesser extent on variation in desaturase activity. PMID- 14740844 TI - Effects of pH and concentrations of linoleic and linolenic acids on extent and intermediates of ruminal biohydrogenation in vitro. AB - Three experiments were conducted by in vitro incubations in ruminal fluid to investigate the effects of pH and amounts of linoleic and linolenic acids on the extent of their biohydrogenation, the proportions of conjugated linoleic acid (CLA) and trans-C18:1 as intermediates, and the ratio trans-10:trans-11 intermediates. The effects of pH and amount of linoleic acid were investigated in kinetic studies, and effects of the amount of linolenic acid were studied with 6 h incubations. With identical initial amounts of linoleic acid, its disappearance declined when the mean pH during incubation was under 6.0 compared with a mean pH over 6.5, and when the amount of linolenic acid increased from 10 to 180 mg/160 ml flask, suggesting an inhibition of the isomerization step of the biohydrogenation. Low pH decreased the ratio of trans-10:trans-11 intermediates. With initial amounts of linoleic acid increasing from 100 to 300 mg, the percentage of linoleic acid disappearance declined, but the amount that disappeared increased, without modification of the trans-10:trans-11 ratio, suggesting a maximal capacity of isomerization rather than an inhibition. Moreover, increasing initial linoleic acid resulted in high amounts of trans C18:1 and an increase of C18:0 that was a linear function of time, suggesting a maximal capacity for the second reduction step of biohydrogenation. High amounts of initial linolenic acid did not affect the amounts of CLA, trans-C18:1, or the ratio trans-10:trans-11. Based on these experiments, a ruminal pH near neutrality with high amount of dietary linoleic acid should modulate the reactions of biohydrogenation in a way that supports CLA and trans-11C18:1 in the rumen. PMID- 14740845 TI - Effects of mannan oligosaccharide or antibiotics in neonatal diets on health and growth of dairy calves. AB - Seventy-two Holstein calves were used to study the effect of feeding antibiotics or mannan oligosaccharides (MOS) in milk replacer. Calves were fed a 20% protein, 20% fat milk replacer containing antibiotics (400 g/ton neomycin + 200 g/ton oxytetracycline), MOS (4 g of Bio-Mos/d), or no additive (control) for 5 wk. Milk replacer was reconstituted to 12.5% dry matter and fed at 12% of birth weight during wk 1 and 14% of birth weight in wk 2 to 5. Fecal scores were monitored 3 times per week; body weight, heart girth, withers height, hip height, and hip width were measured at birth and weekly to 6 wk of age. Addition of MOS or antibiotics increased the probability of normal scores for fecal fluidity, scours severity, and fecal consistency as compared to control calves during the course of the study. Consumption of calf starter increased at a faster rate in calves fed MOS, and these calves consumed more calf starter after weaning (wk 6), than those fed antibiotic. No treatment differences in growth measures, total blood protein, or blood urea nitrogen were detected during the trial. Addition of MOS or antibiotics to milk replacer improved fecal scores in calves. Feed intake was improved in MOS-fed calves compared to antibiotic-fed calves, but this difference did not result in growth differences during the experimental period. The results suggest that antibiotics in milk replacers can be replaced with compounds such as mannan oligosaccharides to obtain similar calf performance. PMID- 14740846 TI - Detection of transgenic and endogenous plant DNA in rumen fluid, duodenal digesta, milk, blood, and feces of lactating dairy cows. AB - The objective was to determine the presence or absence of transgenic and endogenous plant DNA in ruminal fluid, duodenal digesta, milk, blood, and feces, and if found, to determine fragment size. Six multiparous lactating Holstein cows fitted with ruminal and duodenal cannulas received a total mixed ration. There were two treatments (T). In T1, the concentrate contained genetically modified (GM) soybean meal (cp4epsps gene) and GM corn grain (cry1a[b] gene), whereas T2 contained the near isogenic non-GM counterparts. Polymerase chain reaction analysis was used to determine the presence or absence of DNA sequences. Primers were selected to amplify small fragments from single-copy genes (soy lectin and corn high-mobility protein and cp4epsps and cry1a[b] genes from the GM crops) and multicopy genes (bovine mitochondrial cytochrome b and rubisco). Single-copy genes were only detected in the solid phase of rumen and duodenal digesta. In contrast, fragments of the rubisco gene were detected in the majority of samples analyzed in both the liquid and solid phases of ruminal and duodenal digesta, milk, and feces, but rarely in blood. The size of the rubisco gene fragments detected decreased from 1176 bp in ruminal and duodenal digesta to 351 bp in fecal samples. PMID- 14740847 TI - Short communication: Diurnal feeding pattern of lactating dairy cows. AB - The objectives of this research were to: 1) describe the diurnal variation in feed alley attendance patterns of lactating dairy cows, 2) describe the sources of variation in these patterns, and 3) determine the effects of altering the feed push-up schedule on these patterns. An electronic monitoring system was used to record individual cow presence (6-s resolution) at the feed alley for 24 cows housed in a free-stall barn. Cows were subjected to 2 feeding schedules: 1) baseline schedule, where cows were fed at 0600 and 1515 h and feed was pushed closer to the cows at 1100 and 2130 h; and 2) experimental schedule, where 2 additional feed push-ups at 0030 and 0330 h were added to the baseline schedule. With the data collected from the monitoring system, description of the feed alley attendance patterns on a per minute basis of the group of cows was undertaken. Feed alley attendance was consistently higher during the day and early evening compared with the late night and early morning hours. The greatest percentage of cows attending the feed alley was seen after the delivery of fresh feed and the return from milking. The addition of extra feed push-ups in the early morning hours did little to increase feeding activity. It can be concluded that milking and delivery of fresh feed had a much greater affect on the diurnal pattern of feed alley attendance than did the feed push-ups. PMID- 14740848 TI - Impact of type traits on functional herd life of Quebec Holsteins assessed by survival analysis. AB - Survival analysis was used to study the effects of composite and descriptive linear type traits on functional herd life of Quebec Holsteins. Functional herd life was defined as the length of life from first calving to death, culling, or censoring, and was adjusted for 305-d milk production. The dataset contained information from 331,105 cows from Quebec province calving for the first time between 1981 and 1995; 58% of the records had type information. Weibull models were fitted to analyze the data. The hazard function was described as the product of a baseline hazard function and the time-independent effects of age at first calving and type, and the time-dependent effects of year of calving, stage of lactation x lactation number, annual change in herd size, 305-d milk production, and herd-year (random). Analyses were done one at a time for each type trait. The strongest relationships between survival and composite type traits were found for final score, mammary system, and feet and legs. Among the linear type traits, the highest impact on functional herd life was found for traits related to the udder. PMID- 14740849 TI - Short communication: Validation of two animal models for estimation of genetic trends for female fertility in Norwegian dairy cattle. AB - Two animal models were compared with respect to potential bias in genetic trend estimates for female fertility and for their predictive ability. In addition to either a fixed effect for month of first insemination or for month-year of first insemination, the models had fixed effects of age and double insemination and random effects of herd-year and animal. The model with a fixed effect of month of first insemination had a larger positive genetic trend for 56-d nonreturn rate in virgin heifers (0.16% yr), smaller downward bias, and somewhat higher predictive ability. These results demonstrate the importance of verifying models to be used in the calculation of breeding values. PMID- 14740850 TI - Genetic evaluation of fertility using direct and correlated traits. AB - Poor fertility has become a major reason for involuntary culling of dairy cows in the United Kingdom. Calving interval (CI) and body condition score (BCS) are recorded, heritable, genetically correlated with each other, and could be used to extend the scope of dairy indices to include fertility traits. The use of U.K. insemination information for the evaluation of fertility has not been examined previously. Fertility and correlated traits were examined using nationally recorded milk (MILK = daily milk yield at test nearest d 110), BSC, and fertility traits (CI and the insemination traits of nonreturn rate after 56 d, NR56; days to first service, DFS; and number of inseminations per conception, INS). Genetic parameters for the traits were estimated simultaneously with a multitrait sire maternal grandsire (MGS) model and a multitrait BLUP sire MGS model was used to predict sire predicted transmitting abilities for each trait. The relationship between the fertility traits and other predicted transmitting abilities calculated in the United Kingdom was then examined. Heritabilities for the fertility traits were CI = 0.033 +/- 0.01, DFS = 0.037 +/- 0.01, NR56 = 0.018 +/- 0.001, and INS = 0.020 +/- 0.001, with a genetic correlation of 0.671 +/- 0.063 between CI and DFS and -0.939 +/- 0.031 between NR56 and INS. There was an unfavorable genetic correlation between the fertility traits and milk yield and BCS. Predicted transmitting abilities produced are similar in size and range to those produced in other studies and genetic trends are as expected. Results to date are encouraging and suggest that the planned program of work will lead to a fertility index that, when used by breeding companies, will lead to improvements in national dairy cow fertility. PMID- 14740851 TI - Genetic analysis of somatic cell score in Norwegian cattle using random regression test-day models. AB - The dataset used in this analysis contained a total of 341,736 test-day observations of somatic cell scores from 77,110 primiparous daughters of 1965 Norwegian Cattle sires. Initial analyses, using simple random regression models without genetic effects, indicated that use of homogeneous residual variance was appropriate. Further analyses were carried out by use of a repeatability model and 12 random regression sire models. Legendre polynomials of varying order were used to model both permanent environmental and sire effects, as did the Wilmink function, the Lidauer-Mantysaari function, and the Ali-Schaeffer function. For all these models, heritability estimates were lowest at the beginning (0.05 to 0.07) and higher at the end (0.09 to 0.12) of lactation. Genetic correlations between somatic cell scores early and late in lactation were moderate to high (0.38 to 0.71), whereas genetic correlations for adjacent DIM were near unity. Models were compared based on likelihood ratio tests, Bayesian information criterion, Akaike information criterion, residual variance, and predictive ability. Based on prediction of randomly excluded observations, models with 4 coefficients for permanent environmental effect were preferred over simpler models. More highly parameterized models did not substantially increase predictive ability. Evaluation of the different model selection criteria indicated that a reduced order of fit for sire effects was desireable. Models with zeroth- or first-order of fit for sire effects and higher order of fit for permanent environmental effects probably underestimated sire variance. The chosen model had Legendre polynomials with 3 coefficients for sire, and 4 coefficients for permanent environmental effects. For this model, trajectories of sire variance and heritability were similar assuming either homogeneous or heterogeneous residual variance structure. PMID- 14740852 TI - Lifetime profit as an individual trait and prediction of its breeding values in Spanish Holstein cows. AB - Genetic parameters for lifetime profit and some productive traits were estimated from records of 42,401 Holstein cows with first calving before May 1996 from Navarra and Basque Autonomous Regions of Spain. Profit from the first, first two, and first three lactations were tested as early measures of profitability. Profit prediction was tested for another population of 2127 cows using selection indexes (Type-Production and economic indexes) and multitrait analysis for directly predicting profit from first-lactation records. High genetic correlations of actual profit with estimated profit from the first two or first three lactation records, (0.97 and 0.99, respectively) suggest that lifetime profit can be accurately estimated from data in second lactation. Profit was positively correlated to production traits (0.79 to 0.83), functional herd life (0.38), mature body weight (0.25), and days in milk (0.35), but genetic correlation was found to be close to zero with calving interval. Complicated relationships among profit and economic traits (i.e., calving interval, days in milk, and functional herd life) were found. Although the correlation between calving interval and profit was near zero, calving interval was the most important trait after production in prediction of sire profit by a stepwise regression analysis. Profit breeding values from multitrait analysis obtained higher correlation (0.48) with actual profit than Spanish official Type-Production index ICO (0.44) and economic index MEG (0.46). A correlation of 0.49 between profit breeding values and the economic index MEG2002, where stature and calving interval were included as new traits, was obtained. PMID- 14740853 TI - Genetic analysis of body condition score in first-parity Danish Holstein cows. AB - The aim of this study was to test whether genetic components for body condition score (BCS) changed during lactation in first-parity Danish Holsteins. Data were extracted from the national conformation scoring system and consisted of 28,948 records from 3894 herds. Cows were scored once during lactation for BCS on a scale from 1 to 9 with increments of 1. The majority of records were made from d 30 to 150 of lactation. Mean BCS was 4.28 +/- 0.98. Body condition score was lowest in wk 8 to 10 from calving. A multivariate sire model with BCS recordings in six lactation stages treated as different traits was used to analyze the data. In addition, a random regression sire model was used to evaluate the changes in BCS as continuous functions of lactation stage. Estimates of heritability from the multivariate approach ranged from 0.14 to 0.29, and the estimated genetic correlations between BCS at different lactation stages were all higher than 0.82. The random regression model was based on Legendre polynomials (LP) specified on days in milk at scoring. To evaluate the change in mean BCS during lactation, the fixed part of the model included a fifth-order LP on the effect of days in milk at scoring. The highest order of fit used for the sire effect was a third-order LP, but based on likelihood ratio tests this could be reduced to a 0 order, i.e., a model with only the intercept term for the sire effect. This means that the genetic variation is constant over the investigated part of the lactation. Therefore, BCS can be considered the same trait during lactation, and a simple sire model can be used for prediction of breeding values. PMID- 14740854 TI - Genetic improvement of mastitis resistance: validation of somatic cell score and clinical mastitis as selection criteria. AB - Mean daughter deviations for clinical mastitis among second-crop daughters were regressed on predicted transmitting abilities for clinical mastitis and lactation mean somatic cell score in first-crop daughters to validate the predictive ability of these traits as selection criteria for reduced incidence of clinical mastitis. A total of 321 sires had 684,897 second-crop daughters, while predicted transmitting abilities were calculated for 2159 sires, based on 495,681 records of first-crop daughters. Predictive ability, as a measure of efficiency of selection, was 23 to 43% higher for clinical mastitis than for lactation mean somatic cell score. Compared to single-trait selection, predictive ability improved 8 to 13% from utilizing information on both traits. The relative weight that should be assigned to standardized predicted transmitting abilities from univariate genetic analyses were 60 to 67% for clinical mastitis and 33 to 40% for lactation mean somatic cell score. No significant nonlinear genetic relationship between the two traits was found. PMID- 14740855 TI - Genetic diversity and background linkage disequilibrium in the North American Holstein cattle population. AB - The objectives of this study were to 1) identify highly heterozygous Holstein bulls that are as unrelated as possible and widely used in the US dairy industry; 2) quantify the level of genetic diversity in US Holsteins; and 3) determine the extent of background linkage disequilibrium (BLD) and disease trait associated linkage disequilibrium (DLD) in the US Holstein population. Twenty-three Holstein bulls that are not closely related but were widely used in the US dairy industry were genotyped for 54 microsatellite loci. The genotyping was performed on automated DNA sequencers (PE Applied Biosystems, CA), following polymerase chain reaction amplification with fluorescent dye-labeled primers. The heterozygosity for the sampled population ranged from 0.43 to 0.80. This wide range of heterozygosity allows selection of the most heterozygous bulls to develop informative families for gene mapping studies. The degree of genetic diversity in this population is significant and allows selection for traits of economic importance. As expected, there is extensive linkage disequilibrium (LD) in the US Holstein population. About half of the syntenic marker pairs presented a typical pattern of LD produced by DLD. Most of the nonsyntenic marker pairs had a typical pattern of LD arising from BLD. These results suggest that the observed LD is not purely due to genetic drift and migration and that a portion might be due to DLD. This raises our hopes of successful fine-localization of genes for complex traits using LD mapping. PMID- 14740856 TI - Results of a producer survey regarding crossbreeding on US dairy farms. AB - Comprehensive surveys were sent to 528 US dairy producers who are currently practicing crossbreeding in their herds. Fifty usable surveys were returned, and the resulting data included qualitative responses regarding facilities, milk recording plans, milk pricing, crossbreeding goals, breed selection, advantages, disadvantages, and future plans. Quantitative variables included producer scores on a 1 to 5 scale for questions regarding ability to fit into the free stalls and milking parlor, milk volume, component percentages, involuntary culling rate, conception rate, calving difficulty, calf mortality, and prices for breeding stock, cull cows, market steers, and bull calves. The most common first generation crosses involved Jersey and Brown Swiss bulls mated to Holstein cows, and backcrosses to one of these parental breeds were most common in the next generation. Producers who responded to this survey desired, and indicated that they achieved, improvements in fertility, calving ease, longevity, and component percentages through crossbreeding. Respondents indicated that crosses involving the Jersey and Brown Swiss breeds had a clear advantage in longevity relative to purebred Holsteins, and conception rates for crosses of Jersey or Brown Swiss sires on Holstein cows were similar to the (high) conception rates typically achieved in purebred Jersey matings. Respondents also indicated that milk composition was improved in the crossbred cattle, but producers cited some difficulties in marketing crossbred breeding stock and bull calves, and noted that the lack of uniformity within the milking herd created management challenges. Based on results of this survey, it appears that crossbreeding can improve the health, fertility, longevity, and profitability of commercial dairy cattle. However, further research is needed regarding specific heterosis estimates for functional traits in crosses involving each of the major dairy breeds, and improvements are needed in systems for recording the ancestry and breed composition of crossbred animals. PMID- 14740857 TI - Estimation of the protein content of US imports of milk protein concentrates. AB - Recent declines in milk prices in the United States have sparked renewed concern that imports of milk protein concentrates (MPC) are increasingly entering the United States with very low tariff rates and is having an adverse impact on the US dairy industry. Milk protein concentrates are used in the United States in many different products, including the starter culture of cheese, or in nonstandard cheeses such as baker's cheese, ricotta, Feta and Hispanic cheese, processed cheese foods, and nutritional products. One of the difficult aspects of trying to assess the impact of MPC imports on the US dairy industry is to quantify the protein content of these imports. The protein content of MPC imports typically ranges from 40 to 88%. The purpose of this study is to develop a methodology that can be used to estimate the protein content of MPC on a country by country basis. Such an estimate would not only provide information regarding the quantity of protein entering the United States, but would also provide a profile of low- and high-value MPC importers. This is critical for market analysis, since it is the lower valued MPC imports that more directly displaces US-produced skim milk powder. PMID- 14740859 TI - Automatic milking systems, farm size, and milk production. AB - Automatic milking systems (AMS) offer relief from the demanding routine of milking. Although many AMS are in use in Europe and a few are used in the United States, the potential benefit for American farms is uncertain. A farm-simulation model was used to determine the long-term, whole-farm effect of implementing AMS on farm sizes of 30 to 270 cows. Highest farm net return to management and unpaid factors was when AMS were used at maximal milking capacity. Adding stalls to increase milking frequency and possibly increase production generally did not improve net return. Compared with new traditional milking systems, the greatest potential economic benefit was a single-stall AMS on a farm size of 60 cows at a moderate milk production level (8600 kg/cow). On other farm sizes using single stall type robotic units, losses in annual net return of 0 dollars to 300 dollars/cow were projected, with the greatest losses on larger farms and at high milk production (10,900 kg/cow). Systems with one robot serving multiple stalls provided a greater net return than single-stall systems, and this net return was competitive with traditional parlors for 50- to 130-cow farm sizes. The potential benefit of AMS was improved by 100 dollars/cow per year if the AMS increased production an additional 5%. A 20% reduction in initial equipment cost or doubling milking labor cost also improved annual net return of an AMS by up to 100 dollars/cow. Annual net return was reduced by 110 dollars/cow, though, if the economic life of the AMS was reduced by 3 yr for a more rapid depreciation than that normally used with traditional milking systems. Thus, under current assumptions, the economic return for an AMS was similar to that of new parlor systems on smaller farms when the milking capacity of the AMS was well matched to herd size and milk production level. PMID- 14740858 TI - A field study of dietary interactions with monensin on milk fat percentage in lactating dairy cattle. AB - Ninety-one Ontario Holstein dairy herds were surveyed about their lactating cow ration and use of a premix containing monensin to identify possible dietary interactions with monensin on milk fat suppression. All herds were enrolled in Ontario Dairy Herd Improvement (DHI) milk recording, and results from four DHI tests were used. Herd mean fat tests were calculated only for cows between 100 and 200 d in milk to avoid potential confounding due to stage of lactation. Wet forage and total mixed ration (TMR) samples from all herds were evaluated for particle size using the Penn State Particle Size Separator. Of the herds using monensin (n = 58), the dose (per kg of dry matter) ranged from 9 to 14 mg/kg in TMR-fed herds and from approximately 9 to 23 mg/kg in herds in which concentrates were fed separately from forages (component-fed). Of the samples submitted for particle size evaluation, 15% of the haylage (n = 80), 14% of the corn silage (n = 79), and 42% of the TMR (n = 58) samples were classified as having low fiber. There was a significant negative univariable association between monensin and mean milk fat percentage. Monensin significantly reduced milk fat percentage in TMR-fed but not component-fed herds. Fiber length significantly interacted with monensin in TMR-fed herds: Herds that had low fiber in their TMR (< or = 6.0% in the top screen) were susceptible to milk fat decrease by monensin, whereas herds that had adequate fiber (> 6.0%) were not. Monensin also significantly reduced milk fat percentage in herds receiving diets low in nonstructural carbohydrate (< 40.2%) but not in those receiving diets high in NSC (> or = 40.2%). The results of this study suggest that there are significant interactions between monensin and certain dietary factors on milk fat suppression in Holstein dairy herds. PMID- 14740860 TI - Whole-farm nitrogen balance on western dairy farms. AB - Environmental legislation has made it necessary for livestock producers to be able to quantify and adjust the N balance on their farms. Whole-farm N balance and efficiencies were computed for 41 commercial dairies in Utah and Idaho using the University of Maryland Nutrient Balancer. The average N balance, or unaccounted for N, was 81 tonnes per year for the average herd size of 466 cows with 35.8% of the inputs accounted for in the outputs. The major inputs for farms that grew crops (n = 23, herd size = 284 total cows) were imported feed (57.4% of all inputs) and nitrogen fixation (30% of inputs). The major outputs were animal products (primarily milk and some meat, 80% of outputs). For farms that grew no crops (n = 18, herd size = 700 total cows), 98% of the inputs were from imported feed. Of the outputs, 57% of the N was in animal products and 42.9% in manure and compost. Whole-farm balance per product for those farms that grew crops was most affected by herd N utilization efficiency (kg feed N per kg product N), crop N utilization efficiency, and availability of manure N applied to crops, while manure N storage efficiency was of lesser importance. For farms that grew no crops, whole-farm N balance per product was most affected by herd N utilization efficiency and manure N storage efficiency. Maximizing conversion of feed N to product N was the best way to reduce whole-farm N balance. PMID- 14740861 TI - ADSA presidential report. PMID- 14740862 TI - Nursing assessment--taking the history. PMID- 14740863 TI - What is the role of the non-nurse case manager? PMID- 14740864 TI - Research utilization--putting the research evidence into practice. PMID- 14740865 TI - Nurses' perception of their work environment, health, and well-being: a qualitative perspective. AB - The purpose of this analysis was to identify themes nurses expressed in open ended comments at the end of a working conditions survey related to their work environment, health, and well-being. The nursing shortage, downsizing, and long working hours create challenges for nurses trying to deliver quality client care. In addition, nurses are experiencing high levels of physical injury in their work environments. Injuries on the job have led nurses to leave the workplace. Free form comments offered at the completion of a mailed survey of RNs were analyzed for content. Randomly selected nurses from two U.S. states were surveyed in 1999 and 2000 about their jobs with special reference to neck, shoulder, and back pain and disorders. Of the 1,428 respondents, 309 produced usable comments for this content analysis. Constant comparative analysis was used to identify themes present in themes addressed in the nurses' comments included excessive work demands, injustice or unfairness, and nurses' personal solutions to their work environments. Based on the issues raised in the themes, recommendations are provided for improving retention while promoting nurses' health and well-being. PMID- 14740866 TI - Workplace violence in long haul trucking: occupational health nursing update. AB - Almost 2 million workdays and millions of dollars are lost annually because of non-fatal assaults suffered at the workplace (NIOSH, 1996). Twenty workers, on average, are murdered each week in the United States and an estimated 18,000 workers per week are victims of non-fatal assault (NIOSH, 2001). Violence and stress are two interrelated issues that affect the work force. In-depth studies of these issues have not been conducted with long haul truckers in general, or with women in non-traditional, male dominated fields such as the long haul trucking industry. Epidemiological data related to violence and stress experienced by these under-studied populations are needed to plan effective interventions to reduce occupational risks. Studies employing both qualitative and quantitative methods are needed to articulate risk and protective factors related to violence against workers (Runyan, 2001). Occupational health nurses are qualified to participate in the development and implementation of research and intervention studies to improve worker safety related to violence at the workplace for men and women in both traditional and non-traditional occupational roles. PMID- 14740867 TI - Travel health nursing: expanding horizons for occupational health nurses. AB - Pre-travel care, with its focus on disease prevention and health promotion, is the cornerstone of travel health nursing practice. Pre-travel nursing services include risk assessment, immunizations, health counseling, and referrals. Individual travel health and safety risks vary and depend upon a number of different factors. These include: traveler health status, destination, trip duration, season of travel, type of accommodation, trip activities, and prevention efforts. More than 35% of U.S. international travel is business travel. Occupational health nurses have an important role to play to keep traveling employees, including frequent flyers, long stay expatriates, and special needs travelers, healthy, safe, and productive in the "global marketplace." PMID- 14740868 TI - The obesity epidemic. PMID- 14740869 TI - Association of Fcgamma receptor IIa (CD32) polymorphism with severe malaria in West Africa. AB - Malaria continues to claim the lives of more children worldwide than any other infectious disease, and improved understanding of disease immunology is a priority for the development of new therapeutic and vaccination strategies. FcgammaRIIa (CD32) contains a polymorphic variant (H/R131) that has been associated with variability in susceptibility to both bacterial diseases and Plasmodium falciparum parasitemia. We investigated the role of this polymorphism in West Africans with mild and severe malarial disease. The HH131 genotype was significantly associated with susceptibility to severe malaria (P = 0.03, odds ratio = 1.40, 95% confidence interval = 1.02-1.91). In contrast to studies of parasitemia, the presence of the R131 allele, rather than the RR131 genotype, appeared to be the important factor in protection from disease. This is the first evidence for an association between CD32 polymorphism and severe malaria and provides an example of balancing selective pressures from different infectious diseases operating at the same genetic locus. PMID- 14740870 TI - Inducible nitric oxide synthase (NOS2) promoter CCTTT repeat polymorphism: relationship to in vivo nitric oxide production/NOS activity in an asymptomatic malaria-endemic population. AB - Polymorphisms in the inducible nitric oxide synthase gene (NOS2) promoter have been associated with clinical outcome from malaria. These include a CCTTT repeat (CCTTTn) 2.5 kilobases upstream from the NOS2 transcription start site, and two single nucleotide substitutions: G-->C at position -954 (G-954C), and C-->T at position -1173 (C-1173T). Although hypothesized to influence NO production in vivo, the functional relevance of (CCTTT)n and G-954C is uncertain because disease association studies have yielded inconsistent results. This study found no association between CCTTT repeat number and levels of plasma NO metabolites or peripheral blood mononuclear cell NOS activity in a cohort of asymptomatic malaria-exposed coastal Papua New Guineans 1-60 years old. This suggests that (CCTTT)n does not independently influence NOS2 transcription in vivo. Neither the G-954C nor the C-1173T polymorphisms were identified in this population, indicating the variability and complexity of selection for NOS2 promoter polymorphisms in different malaria-endemic populations. PMID- 14740871 TI - Malaria-associated cytokine changes in the placenta of women with pre-term deliveries in Yaounde, Cameroon. AB - The prevalence of pre-term deliveries (PTDs) is increased in women who become infected with Plasmodium falciparum during pregnancy. Because prematurity is a risk factor for newborns, it is important to identify conditions that contribute to malaria-associated PTDs. Plasmodium falciparum-infected erythrocytes sequester in the placenta and attract activated mononuclear cells that secrete pro inflammatory cytokines. Increased inflammatory cytokine levels in other microbial infections are associated with PTDs. To determine if such is the case in women with placental malaria, concentrations of interferon-gamma (IFN-gamma), tumor necrosis factor-alpha (TNF-alpha), interleukin-4 (IL-4), and IL-10 were measured in placental plasma of 391 malaria-infected and -uninfected Cameroonian women with premature and full-term deliveries. Risk factors for malaria-associated PTDs included peripheral and placental parasitemias greater than 1%, maternal anemia, elevated IL-10 levels, and low TNF-alpha:IL-10 ratios due to over-expression of IL-10. Alterations in cytokine levels may contribute to PTDs through the induction of anemia and/or altering cellular immune responses required for eliminating placental parasites. PMID- 14740872 TI - The impact of repeated malaria attacks on the school performance of children. AB - The impact of repeated malarial infections on the school performance of children was investigated in 571 school children 6-14 years of age in a malaria-endemic area in southern Sri Lanka where both Plasmodium falciparum and P. vivax infections are prevalent. Malaria infections confirmed by microscopy were monitored over a six-year period. School performance was assessed by two specially designed, school grade-specific, test papers for Sinhala language and mathematics. The scores for Sinhala language and mathematics for each school term test for the year 1997 were obtained. Malarial infections were a major predictor of children's performance in language and mathematics after controlling for parent's education, monthly family income, and house type. The education of the father predicted language scores but not mathematics scores. A child who experienced more than five attacks of malaria scored approximately 15% less than a child who experienced less than three attacks of malaria. The data suggest that repeated attacks of malaria have an adverse impact on the school performance of children. PMID- 14740873 TI - Evaluation of the Binax NOW ICT test versus polymerase chain reaction and microscopy for the detection of malaria in returned travelers. AB - Microscopic detection of Plasmodium species has been the reference standard for the diagnosis of malaria for more than a century. However, maintaining a sufficient level of expertise in microscopic diagnosis can be challenging, particularly in non-endemic countries. The objective of this study was to compare a new rapid malaria diagnostic device (NOW ICT Malaria Test; Binax, Inc., Portland, ME) to polymerase chain reaction (PCR) and expert microscopy for the diagnosis of malaria in 256 febrile returned travelers. Compared with PCR, the NOW ICT test showed a sensitivity of 94% for the detection of P. falciparum malaria (96% for pure P. falciparum infection) and 84% for non-P. falciparum infections (87% for pure P. vivax infections and 62% for pure P. ovale and P. malariae infections), with an overall specificity of 99%. The Binax NOW ICT may represent a useful adjunct for the diagnosis of P. falciparum and P. vivax malaria in febrile returned travelers. PMID- 14740874 TI - Adaptation of a strain of Plasmodium falciparum from Ghana to Aotus lemurinus griseimembra, A. nancymai, and A. vociferans monkeys. AB - A strain of Plasmodium falciparum from Ghana was adapted to Aotus lemurinus griseimembra, A. nancymai, and A. vociferans monkeys. Gametocytes in splenectomized A. nancymai were infective to Anopheles freeborni mosquitoes. Sporozoite transmission was accomplished in two splenectomized A. nancymai with prepatent periods of 22 and 25 days. The Ghana III/CDC strain of P. falciparum is susceptible to treatment with chloroquine and mefloquine. PMID- 14740875 TI - Prediction of Plasmodium falciparum resistance to sulfadoxine/pyrimethamine in vivo by mutations in the dihydrofolate reductase and dihydropteroate synthetase genes: a comparative study between sites of differing endemicity. AB - Plasmodium falciparum resistance to sulfadoxine/pyrimethamine (S/P) is due to mutations in the dihydrofolate reductase (dhfr) and dihydropteroate synthetase (dhfr) genes. Large-scale screening of the prevalence of these mutations could facilitate the surveillance of the level of S/P resistance in vivo. The prevalence of mutations in dhfr and dhps in relation to S/P efficacy was studied in four sites of differing endemicity in Sudan, Mozambique, and Tanzania. The sites were organized in order of increasing resistance and a significant increase in the prevalence of triple mutations in codons c51, c59, and c108 of dhfr was observed. A similar trend was observed when dhfr genotypes were combined with c437 of dhps. Since the differences in S/P resistance between the sites were minor, but nevertheless revealed major differences in dhfr genotype prevalence, the role of dhfr as a general molecular marker seems debatable. The differences may reflect variation in the duration and magnitude of S/P usage (or other antifolate drugs) between the sites. Thus, triple dhfr mutations may prove suitable only as a general guideline for detecting emerging S/P resistance in areas where S/P has been introduced recently. However, changes in susceptibility within the same area with moderate levels of resistance may be possible by longitudinal surveillance of a subset of dhfr/dhps mutations that has been associated with S/P resistance in vivo in a defined location. PMID- 14740876 TI - High seasonal variation in entomologic inoculation rates in Eritrea, a semi-arid region of unstable malaria in Africa. AB - Entomologic studies were conducted in eight villages to investigate the patterns of malaria transmission in different ecologic zones in Eritrea. Mosquito collections were conducted for 24 months between September 1999 and January 2002. The biting rates of Anopheles arabiensis were highly seasonal, with activity concentrated in the wet season between June and October in the highlands and western lowlands, and between December and March in the coastal region. The biting rates in the western lowlands were twice as high as in the western escarpment and 20 times higher than in the coastal region. Sporozoite rates were not significantly different among villages. The risk of infection ranged from zero on the coast to 70.6 infective bites per year in the western lowlands. The number of days it would take for an individual to receive an infective bite from an infected An. arabiensis was variable among villages (range = 2.8-203.1 days). The data revealed the presence of only one main malaria transmission period between July and October for the highlands and western lowlands. Peak inoculation rates were recorded in August and September (range = 0.29-43.6 infective bits/person/month) at all sites over the two-year period. The annual entomologic inoculation rates (EIRs) varied greatly depending on year. The EIR profiles indicated that the risk of exposure to infected mosquitoes is highly heterogeneous and seasonal, with high inoculation rates during the rainy season, and with little or no transmission during the dry season. This study demonstrates the need to generate spatial and temporal data on transmission intensity on smaller scales to guide targeted control of malaria operations in semi-arid regions. Furthermore, EIR estimates derived in the present study provide a means of quantifying levels of exposure to infected mosquitoes in different regions of the country and could be important for evaluating the efficacy of vector control measures, since Eritrea has made significant steps in reducing the burden of malaria based on the Roll Back Malaria initiative of the World Health Organization. PMID- 14740877 TI - Frequency distribution of antimalarial drug-resistant alleles among isolates of Plasmodium falciparum in Purworejo district, Central Java Province, Indonesia. AB - Treatment failures to the first- and second-lines antimalarial drugs chloroquine and sulfadoxine-pyrimethamine have increased in the Purworejo district on the island of Java, Indonesia. A molecular epidemiologic study was conducted to determine the frequency distribution of mutant alleles of the genes associated with the resistance among the isolates of Plasmodium falciparum from the area. Analyses using a polymerase chain reaction and restriction fragment length polymorphism showed that nearly all of the 111 samples carried mutant alleles in genes associated with chloroquine resistance: P. falciparum multi-drug resistance 1 (pfmdrl) 86Y (92%), 1042D (4.5%), and P. falciparum chloroquine resistance transporter (pfcrt) 76T (99.1%). Mutant alleles of the in the dihydrofolate reductase (dhfr) gene were also high (84.7%), either as 108N and 108T or paired with 59R, and 16V, respectively. Mutant alleles in the dihydropteroate synthase gene were the least common, either as a single 437G mutation (35.3%) or paired with 540E (26.5%). These results are consistent with the antimalarial drug resistance situation in the area and emphasize the need for a proper treatment strategy. PMID- 14740878 TI - Adverse reactions to ivermectin treatment in Simulium neavei-transmitted onchocerciasis. AB - To assess adverse effects of ivermectin treatment in a Simulium neavei transmitted focus of onchocerciasis, a study was conducted with 1,246 patients infected with Onchocerca volvulus in eight villages in western Uganda. Study participants were treated the first time with a single dose of 150 microg/kg of ivermectin. Adverse reactions to ivermectin were determined through questioning and clinical examination during house-to-house visits to the participants within 48 hours after ivermectin treatment. Overall adverse reactions were observed in 737 (59.1%) patients. Severe reactions were rare (10 patients, 1.4%). Our data show that adverse reactions to ivermectin in an S. neavei-transmitted onchocerciasis focus in western Uganda occur frequently. In spite of the fact that many patients showed adverse reactions to ivermectin, the drug was well accepted and appreciated by the population. PMID- 14740879 TI - Visceral leishmaniasis: consequences of a neglected disease in a Bangladeshi community. AB - Visceral leishmaniasis, or kala azar (KA), affects the rural poor, causing significant morbidity and mortality. We examined the epidemiologic, social, and economic impact of KA in a village in Bangladesh. A population-based survey among 2,348 people demonstrated a KA incidence of 2% per year from 2000 to 2002, with a case-fatality rate of 19% among adult women, compared with 6-8% among other demographic groups. Kala azar cases were geographically clustered in certain sections of the village. Anti-leishmanial drug shortages and the high cost of diagnosis and treatment caused substantial emotional and economic hardship for affected families. Communities wanted to learn more about KA, and were willing to take collective action to confront the problems it causes. To decrease the KA burden in endemic areas, community efforts should be supplemented with effective treatment programs to ensure access to appropriate and affordable diagnosis and case management. PMID- 14740880 TI - Risk factors for infection during a severe dengue outbreak in El Salvador in 2000. AB - In 2000, El Salvador experienced a large dengue-2 virus epidemic with many severe cases. A seroepidemiologic survey was conducted in 106 randomly selected households (501 residents) in an affected community (Las Pampitas). The frequency of recent infection, documented by the presence of IgM antibodies or high-titer IgG antibodies to dengue virus, was estimated to be 9.8% (95% confidence interval [CI] = 5.8-13.7), of which at least 44% were secondary infections. The number of containers positive for Aedes mosquito larvae or pupae per 100 premises (Breteau Index) was 62 and the frequency of positive premises (House Index) was 36%; 33% (35 of 106) of the informants reported having taken action against mosquito larval habitats and 82% (87 of 106) reported having taken actions against adult mosquitoes. Recent infection was associated with the presence in the home environment of mosquito infested discarded cans (odds ratio [OR] = 4.30, 95% CI = 2.54-7.28), infested discarded plastic containers (OR = 3.98, 95% CI = 1.05 15.05), and discarded tire casings (OR = 2.57, 95% CI = 1.09-6.04). The population attributable fractions associated with these factors were 4%, 13%, and 31%, respectively. Our data suggest that targeted community cleanup campaigns, particularly those directed at discarded tires and solid waste, are likely to have the greatest impact on reducing the risk of dengue infection. PMID- 14740881 TI - Use of a geographic information system for defining spatial risk for dengue transmission in Bangladesh: role for Aedes albopictus in an urban outbreak. AB - We used conventional and spatial analytical tools to characterize patterns of transmission during a community-wide outbreak of dengue fever and dengue hemorrhagic fever in Dhaka, Bangladesh in 2000. A comprehensive household-level mosquito vector survey and interview was conducted to obtain data on mosquito species and breeding as well as illness consistent with dengue. Clusters of dengue illnesses and high-density vector populations were observed in a distinct sector of the city. Dengue clusters are less identifiable in areas further away from major hospitals, suggesting that proximity to hospitals determines whether cases of dengue are diagnosed. Focusing on those areas relatively close to hospitals, we found a spatial association between dengue clusters and vector populations. Households reporting a recent dengue illness were more likely to have Aedes albopictus larvae present in the home when compared with households not reporting cases. Households reporting a recent dengue illness were also more likely to have a neighbor with Ae. albopictus present in the home. In contrast, the presence of Aedes aegypti within the premises as well as the homes of neighbors (within 50 meters) was not associated with dengue illness. Given that the breeding habitats for Ae. albopictus are somewhat distinct from those of Ae. aegypti, the findings of this study have implications for control of dengue transmission in this urban setting where much of the focus has been on indoor mosquito breeding and transmission. Public health officials may find the disease environment map useful for planning targeted interventions because it displays areas where transmission is most intense. PMID- 14740882 TI - Avian host preference by vectors of eastern equine encephalomyelitis virus. AB - An important variable in the amplification and escape from the enzootic cycle of the arboviral encephalitides is the degree of contact between avian hosts and mosquito vectors. To analyze this interaction in detail, blood-fed mosquitoes that were confirmed vectors of eastern equine encephalomyelitis (EEE) virus were collected in 2002 from an enzootic site in central Alabama during the time this virus was actively transmitted. Avian-derived blood meals were identified to the species level of the host, and the proportion derived from each species was compared with the overall composition of the avifauna at the study site. The EEE vector mosquito species fed significantly more on some bird species and less on other species than expected given the overall abundance, biomass, or surface area of the local avifauna. When viewed collectively, these data suggest that these mosquitoes are selectively targeting particular avian species. PMID- 14740883 TI - Short report: detection of Japanese encephalitis virus in mouse peripheral blood mononuclear cells using an in situ reverse transcriptase-polymerase chain reaction. AB - Japanese encephalitis (JE) is an important mosquito-borne viral disease in Southeast Asia. Isolation of JE virus from peripheral blood is usually difficult because of transient and low titer of viremia. An in situ reverse transcriptase polymerase chain reaction (RT-PCR) method was designed to amplify gene (envelope) fragments of JE virus residing in peripheral blood mononuclear cells (PBMCs) without extraction of RNA. Baby hamster kidney-21 cells infected with the T1P1 strain of JE virus (an isolate from Armigeres subalbatus collected in Taiwan) were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. The RT-PCR was then performed in microtubes using digoxigenin-labeled primers. Virus positive PBMCs were detected in mice at day 1 and day 3, but not day 5, after intravenous inoculation with JE virus, suggesting that detectable virus circulating in the blood of mice is present for only 2-3 days. On examination of mouse brain tissues, viral RNAs were absent until day 3 post-inoculation. This implied that virus migration from the peripheral blood into the central nervous system occurs at or after day 3 post-inoculation. This method is unique in that the reactions can be conducted in tubes; this makes it convenient, accurate, and efficient compared with the conventional in situ RT-PCR on slides. PMID- 14740884 TI - Modulation of cytokine expression in human keratinocytes and fibroblasts by extracts of scabies mites. AB - Sarcoptes scabiei lives in the stratum corneum of its mammalian host. Keratinocytes and fibroblasts are among the first cells to encounter the burrowing mite and its products. The aim of this study was to determine if molecules in an extract of S. scabiei modulate the expression of cytokines by keratinocytes and fibroblasts. Human keratinocytes and fibroblasts were exposed to an extract of S. scabiei var. canis in the absence or presence of Escherichia coli lipopolysaccharide. Cytokine expression was measured by an enzyme-linked immunosorbent assay. Components in the S. scabiei extract induced marked increases in secretion of interleukin-6 (IL-6) and vascular endothelial growth factor (VEGF) and slight increases in production of granulocyte-colony stimulating factor (G-CSF) by keratinocytes. The scabies extract down-regulated keratinocyte secretion of IL-1 receptor antagonist, but did not influence the production of IL-1alpha or IL-1beta. In comparison, components in the scabies extract induced marked increases in the elaboration of IL-6, IL-8, G-CSF, and VEGF by fibroblasts. Neither cell type produced eotaxin, stem cell factor, or tumor necrosis factor-alpha under any of the conditions tested. This study demonstrates that components in an extract of the mite S. scabiei are able to influence cytokine expression by human keratinocytes and fibroblasts. PMID- 14740885 TI - Intestinal parasites among African refugees resettled in Massachusetts and the impact of an overseas pre-departure treatment program. AB - This study analyzed the prevalence of intestinal parasitoses diagnosed shortly after arrival in the United States among African refugees before and after implementation of an overseas program of empirical treatment with albendazole. Variables included results of microscopy of a single stool specimen, age, sex, ethnicity, departure origin, and receipt of albendazole. Of 1,254 refugees, 56% had intestinal parasites. Fourteen percent had helminths, and 2% had multiple helminths. In addition, 52% had protozoans with 25% having multiple protozoans. The most common pathogens were Giardia lamblia (14%) and Trichuris trichiura (9%). Overall, refugees who arrived in Massachusetts after implementation of the treatment program were less likely to have any parasites (odds ratio [OR] = 0.61, 95% confidence interval [CI] = 0.47-0.78) and helminths (OR = 0.15, 95% CI = 0.09 0.24) than refugees who arrived previously. These more recently arrived refugees were also less likely to have hookworm (OR = 0.03, 95% CI = 0.00-0.29), Trichuris (OR = 0.05, 95% CI = 0.02-0.13), Ascaris (OR = 0.07, 95% CI = 0.01-0.58), and Entamoeba histolytica (OR = 0.47, 95% CI = 0.26-0.86). Empirically treating refugees prior to departure for the United States appears to have resulted in decreases in intestinal helminths and possibly some protozoans among African refugees tested shortly after arrival in this country. PMID- 14740886 TI - A large outbreak of histoplasmosis among American travelers associated with a hotel in Acapulco, Mexico, spring 2001. AB - During spring 2001, college students from Pennsylvania reported an acute febrile respiratory illness after returning from spring break vacation in Acapulco, Mexico. Acute pulmonary histoplasmosis was presumptively diagnosed and the cluster of illness was reported to the Centers of Disease Control and Prevention. A large investigation then ensued, which included finding student-travelers for interviews and requesting sera for histoplasmosis testing. We defined a clinical case by fever and at least one of the following: cough, shortness of breath, chest pain, or headache, in an Acapulco traveler during March-May 2001. A laboratory-confirmed case had positive serology. An initial study determined that the likely site of histoplasmosis exposure was Hotel H; we therefore performed a large cohort study among travelers who stayed at Hotel H. Of 757 contacted, 262 (36%) met the clinical case definition. Of 273 serum specimens tested, 148 (54%) were positive. Frequent use of Hotel H's stairwells, where construction was ongoing, was associated with increased risk of illness (relative risk = 10.5, 95% confidence interval = 3.7-30.5; P < 0.001). This is the first histoplasmosis outbreak associated with a hotel undergoing construction. Hotels in endemic areas should consider construction precaution measures to prevent histoplasmosis among their guests. PMID- 14740887 TI - Mutagenesis studies of the F1F0 ATP synthase b subunit membrane domain. AB - A homodimer of b subunits constitutes the peripheral stalk linking the F1 and F0 sectors of the Escherichia coli ATP synthase. Each b subunit has a single membrane domain. The constraints on the membrane domain have been studied by systematic mutagenesis. Replacement of a segment proximal to the cytoplasmic side of the membrane had minimal impact on F1F0 ATP synthase. However, multiple substitutions on the periplasmic side resulted in defects in assembly of the enzyme complex. These mutants had insufficient oxidative phosphorylation to support growth, and biochemical studies showed little F1F0 ATPase and no detectable ATP-driven proton pumping activity. Expression of the b(N2A,T6A,Q10A) subunit was also oxidative phosphorylation deficient, but the b(N2A,T6A,Q10A) protein was incorporated into an F1F0 complex. Single amino acid substitutions had minimal reductions in F1F0 ATP synthase function. The evidence suggests that the b subunit membrane domain has several sites of interaction contributing to assembly of F0, and that these interactions are strongest on the periplasmic side of the bilayer. PMID- 14740888 TI - Overexpression, purification, and characterization of human and bovine mitochondrial ATPase inhibitors: comparison of the properties of mammalian and yeast ATPase inhibitors. AB - Mitochondrial ATP synthase (F1F0-ATPase) is regulated by an intrinsic ATPase inhibitor protein. In this study, we overexpressed and purified human and bovine ATPase inhibitors and their properties were compared with those of a yeast inhibitor. The human and bovine inhibitors inhibited bovine ATPase in a similar way. The yeast inhibitor also inhibited bovine F1F0-ATPase, although the activity was about three times lower than the mammalian inhibitors. All three inhibitors inhibited yeast F1F0-ATPase in a similar way. The activities of all inhibitors decreased at higher pH, but the magnitude of the decrease was different for each combination of inhibitor and ATPase. The results obtained in this study show that the inhibitory mechanism of the inhibitors was basically shared in yeast and mammals, but that mammalian inhibitors require unique residues, which are lacking in the yeast inhibitor, for their maximum inhibitory activity. Common inhibitory sites of mammalian and yeast inhibitors are suggested. PMID- 14740889 TI - Molecular properties of purified human uncoupling protein 2 refolded from bacterial inclusion bodies. AB - One way to study low-abundance mammalian mitochondrial carriers is by ectopically expressing them as bacterial inclusion bodies. Problems encountered with this approach include protein refolding, homogeneity, and stability. In this study, we investigated protein refolding and homogeneity properties of inclusion body human uncoupling protein 2 (UCP2). N-methylanthraniloyl-tagged ATP (Mant-ATP) experiments indicated two independent inclusion body UCP2 binding sites with dissociation constants (Kd) of 0.3-0.5 and 23-92 microM. Dimethylanthranilate, the fluorescent tag without nucleotide, bound with a Kd of greater than 100 microM, suggesting that the low affinity site reflected binding of the tag. By direct titration, UCP2 bound [8-(14)C] ATP and [8-(14)C] ADP with Kds of 4-5 and 16-18 microM, respectively. Mg2+ (2 mM) reduced the apparent ATP affinity to 53 microM, an effect entirely explained by chelation of ATP; with Mg2+, Kd using calculated free ATP was 3 microM. A combination of gel filtration, Cu2+ phenanthroline cross-linking, and ultracentrifugation indicated that 75-80% of UCP2 was in a monodisperse, 197 kDa form while the remainder was aggregated. We conclude that (a) Mant-tagged nucleotides are useful fluorescent probes with isolated UCP2 when used with dimethylanthranilate controls; (b) UCP2 binds Mg2+ free nucleotides: the Kd for ATP is about 3-5 microM and for Mant-ATP it is about 10 times lower; and (c) in C12E9 detergent, the monodisperse protein may be in dimeric form. PMID- 14740890 TI - Effect of pH on the steady state kinetics of bovine heart NADH: coenzyme Q oxidoreductase. AB - Complete initial steady state kinetics of NADH-decylubiquinone (DQ) oxidoreductase reaction between pH 6.5 and 9.0 show an ordered sequential mechanism in which the order of substrate bindings and product releases is NADH DQ-DQH2-NAD+. NADH binding to the free enzyme is accelerated by protonation of an amino acid (possibly a histidine) residue. The NADH release is negligibly slow under the turnover conditions. The rate of DQ binding to the NADH-bound enzyme and the maximal rate at the saturating concentrations of the two substrates, which is determined by the rates of DQH2 formation in the active site and releases of DQH2 and NAD+ from the enzyme, are insensitive to pH, in contrast to clear pH dependencies of the maximal rates of cytochrome c oxidase and cytochrome bc1 complex. Physiological significances of these results are discussed. PMID- 14740891 TI - Sodium dependency of the photosynthetic electron transport in the alkaliphilic cyanobacterium Arthrospira platensis. AB - Arthrospira (Spirulina) platensis (A. platensis) is a model organism for investigation of adaptation of photosynthetic organisms to extreme environmental conditions: the cell functions in this cyanobacterium are optimized to high pH and high concentration (150-250 mM) of Na+. However, the mechanism of the possible fine-tuning of the photosynthetic functions to these extreme conditions and/or the regulation of the cellular environment to optimize the photosynthetic functions is poorly understood. In this work we investigated the effect of Na ions on different photosynthetic activities: linear electron transport reactions (measured by means of polarography and spectrophotometry), the activity of photosystem II (PS II) (thermoluminescence and chlorophyll a fluorescence induction), and redox turnover of the cytochrome b6f complex (flash photolysis); and measured the changes of the intracellular pH (9-aminoacridine fluorescence). It was found that sodium deprivation of cells in the dark at pH 10 inhibited, within 40 min, all measured photosynthetic reactions, and led to an alkalinization of the intracellular pH, which rose from the physiological value of about 8.3-9.6. These were partially and totally restored by readdition of Na ions at 2.5-25 mM and about 200 mM, respectively. The intracellular pH and the photosynthetic functions were also sensitive to monensin, an exogenous Na+/H+ exchanger, which collapses both proton and sodium gradients across the cytoplasmic membrane. These observations explain the strict Na+-dependency of the photosynthetic electron transport at high extracellular pH, provide experimental evidence on the alkalization of the intracellular environment, and support the hypothesized role of an Na+/H+ antiport through the plasma membrane in pH homeostasis (Schlesinger et al. (1996). J. Phycol. 32, 608-613). Further, we show that (i) the specific site of inactivation of the photosynthetic electron transport at alkaline pH is to be found at the water splitting enzyme; (ii) in contrast to earlier reports, the inactivation occurs in the dark and, for short periods, without detectable damage in the photosynthetic apparatus; and (iii) in contrast to high pH, Na+ dependency in the neutral pH range is shown not to originate from PSII, but from the acceptor side of PSI. These data permit us to conclude that the intracellular environment rather than the machinery of the photosynthetic electron transport is adjusted to the extreme conditions of high pH and high Na+ concentration. PMID- 14740892 TI - Contribution of the phosphorylable complex I in the growth phase-dependent respiration of C6 glioma cells in vitro. AB - The energy metabolism of rat C6 glioma cells was investigated as a function of the growth phases. Three-dimensional cultures of C6 cells exhibited diminished respiration and respiratory capacity during the early growth phase, before reaching confluence. This decrease in respiration was neither due to changes in the respiratory complex content nor in the mitochondrial mass per se. Nevertheless, a quantitative correlation was found between cellular respiration and the rotenone-sensitive NADH ubiquinone oxidoreductase (i.e. complex I) activity. Immunoblot analysis showed that phosphorylation of the 18 kDa-subunit of this complex was associated with the growth-phase dependent modulation of complex I and respiratory activity in C6 cells. In addition, by using forskolin or dibutyryl cAMP, short-term activation of protein kinases A of C6 cells correlated with increased phosphorylation of the 18-kDa subunit of complex I, activated NADH ubiquinone oxidoreductase activity and stimulated cellular respiration. These findings suggest that complex I of C6 glioma cells is a key regulating step that modulates the oxidative phosphorylation capacity during growth phase transitions. PMID- 14740894 TI - Presidential address. Flavor buds and other delights. AB - Using good sense and some biological information, one can enjoy a delightful morsel or enhanced meal from a variety of parasites, either raw or cooked. Bon Appetit! PMID- 14740893 TI - Hypertriglyceridemia increases mitochondrial resting respiration and susceptibility to permeability transition. AB - High plasma level of triglycerides (TGs) is a common feature in atherosclerosis, obesity, diabetes, alcoholism, stress, and infection. Since mitochondria have been implicated in cell death under a variety of metabolic disorders, we examined liver mitochondrial functions in hypertriglyceridemic transgenic mice. Hypertriglyceridemia increased resting respiration and predisposed to mitochondrial permeability transition (MPT). Ciprofibrate therapy reduced plasma TG levels, normalized respiration, and prevented MPT. The higher resting respiration in transgenic mitochondria remained in the presence of the adenine nucleotide carrier inhibitor, carboxyatractyloside, bovine serum albumin, and the uncoupling proteins (UCPs) inhibitor, GDP. UCP2 content was similar in both control and transgenic mitochondria. We propose that faster resting respiration represents a regulated adaptation to oxidize excess free fatty acid in the transgenic mice. PMID- 14740895 TI - Presentation of the 2003 recipient of the American Society of Parasitologists' Mentor Award, Dr. John Janovy, Jr. PMID- 14740896 TI - Acceptance of the Clark P. Read Mentor Award: the Teague Self lessons. PMID- 14740897 TI - Introduction of John R. Barta as the Henry Baldwin Ward medalist for 2003. PMID- 14740898 TI - Acceptance of the 2003 Henry Baldwin Ward Medal. PMID- 14740899 TI - Killing of schistosomes by elastase and hydrogen peroxide: implications for leukocyte-mediated schistosome killing. AB - Activated leukocytes participate in immunity to infection by the parasitic blood fluke Schistosoma mansoni. They attach to the surface of schistosomes and secrete schistosomicidal substances. Cationic proteins, hydrolytic enzymes, and oxidants, produced by the leukocytes, have been implicated in the damage to the schistosomes. To examine the possible involvement of elastase in the killing of schistosomes by leukocytes, young and adult stages of S. mansoni were treated in vitro with pancreatic elastase (PE) and neutrophil elastase (NE). Schistosomula, lung-stage schistosomula (LSS), and adult worms (AW) have been found to be sensitive to both PE and NE. Male AW were more sensitive to PE than female AW. The enzymatic activity of elastase is essential for its toxic effect because heat inactivation and specific elastase inhibitors prevented elastase-mediated schistosome killing. Thus, alpha1-antitrypsin and the chloromethyl ketone (CMK) derived tetrapeptides Ala-Ala-Pro-Val-CMK and Ala-Ala-Pro-Ala-CMK but not Ala-Ala Pro-Phe-CMK and Ala-Ala-Pro-Leu-CMK blocked PE caseinolytic and schistosomulicidal activities. As shown previously, schistosomes are also efficiently killed by hydrogen peroxide. LSS appear to be more resistant than AW and early-stage schistosomula to the lytic effects of hydrogen peroxide. Cotreatment experiments with both elastase and hydrogen peroxide indicated that they exert an additive toxic effect and that hydrogen peroxide sensitizes schistosomula to the toxic effect of elastase but not vice versa. These results demonstrate, for the first time, that elastases may be toxic molecules used by neutrophils, eosinophils, and macrophages to kill various developmental stages of S. mansoni. PMID- 14740900 TI - Guanosine 3',5'-cyclic monophosphate: a tapeworm-secreted signal molecule communicating with the rat host's small intestine. AB - Tapeworms alter the physiological environment of the host's small intestinal lumen by contracting the intestinal smooth muscle, thereby slowing the transit of intestinal contents. We hypothesize that parasite-to-host molecular signaling is responsible for the specific patterns of small intestinal smooth muscle contraction observed both during tapeworm infection and after the infusion of tapeworm-secreted molecules into the intestinal lumen of unanesthetized rats. Of the tapeworm-secreted compounds tested, only lumenal infusion of guanosine 3',5' cyclic monophosphate (cGMP) induced contractile patterns that mimic those observed during tapeworm infection. The response to cGMP occurred in a concentration-dependent fashion. Our study clearly demonstrates that cGMP can serve as an extracellular signal molecule regulating small intestinal motility mechanisms in vivo. PMID- 14740901 TI - Babesia gibsoni-specific isoenzymes related to energy metabolism of the parasite in infected erythrocytes. AB - To clarify the cause of the predilection of Babesia gibsoni for reticulocytes and canine HK erythrocytes (containing high concentrations of potassium) with inherited high concentrations of some amino acids, including glutamate, 4 enzymes in B. gibsoni parasites were examined by polyacrylamide gel electrophoresis (PAGE). The enzymes, i.e., hexokinase, glucose phosphate isomerase, lactate dehydrogenase, and glutamate dehydrogenase (GDH), were found to be associated with B. gibsoni parasites. The parasite-specific enzymes were shown to have different mobility patterns in PAGE from those found in normal canine erythrocytes. GDH, which is able to oxidize glutamate to alpha-ketoglutarate, an intermediate in the citric acid cycle in mitochondria, was detected only in the parasites. Electron microscopy of the parasites revealed double-membraned organelles similar to mitochondria in their cytoplasm. The parasites in in vitro culture contained many more mitochondrialike organelles than those in the peripheral blood of infected dogs. In addition, the size of parasites cultured in vitro was significantly larger than that of parasites in the peripheral blood. Based on these results, it is suggested that B. gibsoni may use glucose as an energy source in its own glycolytic pathway. Moreover, the parasite may also be capable of oxidizing glutamate via GDH in the citric acid cycle, which may operate in the mitochondrialike organelles within the parasite. This may explain the predilection of B. gibsoni for canine reticulocytes and HK erythrocytes with a high concentration of glutamate. PMID- 14740902 TI - Is the prevalence of Taenia taeniaeformis in Microtus arvalis dependent on population density? AB - Populations of common voles Microtus arvalis were studied as hosts of the tapeworm Taenia taeniaeformis during a 14-yr survey. They were monitored in spring, summer, and autumn in a pastoral ecosystem in eastern France. A total of 7,574 voles were sampled during 2 multiannual population fluctuations. A third fluctuation was sampled during the increase phase only. Overall prevalence was lowest in summer (0.6%), increased by 3 times in autumn (1.5%) and a further 5 times in spring (7.8%). Analysis of prevalence, based on 7,384 voles, by multiple logistic regression revealed that extrinsic factors such as season and intrinsic factors such as host age and host density have a combined effect. In the longer term, host age and host density were positively associated with prevalence in summer. Host density was strongly associated with autumn prevalence. There was no association between the fluctuation phase and prevalence. The study shows that a long timescale (here a multiannual survey) is necessary to demonstrate the positive effect of host density on the prevalence of this indirectly transmitted parasite. The demonstration of this relationship depends on the rodents being intermediate rather than definitive hosts. PMID- 14740903 TI - Microparasite assemblages of conspecific shrew populations in southern California. AB - The microparasite component communities of 2 species of shrews, Notiosorex crawfordi and Sorex ornatus, were investigated for the first time in 2 isolated and 3 continuous landscapes in southern California. With microscopical examination, a total of 6 parasite species was found in N. crawfordi and 8 species in S. ornatus. The highest number (5) of parasite species was detected in the lungs. The corrected estimate of parasite species richness did not significantly correlate with the host abundance in either shrew species. Altitude, and also latitude in N. crawfordi, appeared to be significantly positively associated with the parasite species richness, but this could be due to a false association because of the rare occurrence of some of the parasites or the small altitude range (or both). No other landscape variable analyzed (location, size of the study site, disturbance) was significantly associated with the parasite species richness of the shrews. The parasite assemblages of the 2 shrew species were similar despite the fact that N. crawfordi has a lower metabolic rate than S. ornatus. PMID- 14740904 TI - Egg size variability in trematodes: test of the bet-hedging hypothesis. AB - The hypothesis according to which egg size variability in hermaphroditic parasites results from bet-hedging was investigated in a comparative analysis using trematodes as a model. We hypothesized that the species reproducing mainly by self-fertilization should produce smaller eggs than those species that regularly practice cross-fertilization. Indeed, because self-fertilization is usually associated with inbreeding depression, selection should favor individuals spreading the risk of genetically disturbed development across more but smaller eggs, instead of producing fewer eggs, each possessing a large resource supply, of which many may fail to develop because of genetic deficiencies. On the basis of earlier theoretical and empirical studies, we assumed that the ratio length of testis-length of ovary positively correlates with the mating group size and, hence, with opportunities for cross-fertilization. In accordance with the bet hedging hypothesis, we found, across trematode species, a positive relationship between this ratio and the mean egg volume produced by adults. This result was, however, observed only for the trematodes infecting birds and not for the species infecting fishes and mammals. In addition, once the influence of trematode phylogeny was taken into account, there was no significant trend, suggesting that phylogenetic legacies played a large role in generating the previous signal. Experimental tests of the bet-hedging hypothesis will be necessary to clarify the matter. PMID- 14740905 TI - Pterygodermatites nycticebi (Nematoda: Rictulariidae): accidental detection of encapsulated third-stage larvae in the tissue of a white-fronted marmoset. AB - Twin, white-fronted marmosets (Callithrix geoffroyi) born and raised in a zoo in Japan died at 7 mo of age. Several encapsulated nematode larvae were detected in the intestinal wall, as well as a few in the mesenteric lymph nodes of 1 of the twins. In the other marmoset, no encapsulated nematode larva was detected in the organs, but many adult Pterygodermatites nycticebi were found in the intestinal lumen. In the past 5 yr, 5 primates kept in the same zoo, i.e., 1 squirrel monkey (Saimiri sciureus), 2 Pygmy marmosets (Cebuella pygmaea), 1 Senegal galago (Galago senegalensis), and 1 cotton-top tamarin (Saguinus oedipus), died from heavy infestation with the same nematode. A few migrating larvae of the rictulariid were also identified histologically in the intestinal wall and liver of the cotton-top tamarin. Although no other primate currently held in the same zoo was infected with the rictulariid, German cockroaches (Blattella germanica) collected with traps near marmoset cages had encapsulated P. nycticebi larvae, indicating latent perpetuation of the life cycle of this rictulariid species in the zoo premises. Our results indicated that encapsulation or migration of third stage larvae of P. nycticebi might occur accidentally in the organs of callithrichid primates. PMID- 14740906 TI - Reduction in the level of infection of the bivalve Anodonta piscinalis by the copepod Paraergasilus rylovi using high temperature and low oxygen. AB - The aim of this study was to develop a method to kill or expel the gill-dwelling crustacean parasite Paraergasilus rylovi from a common freshwater clam, Anodonta piscinalis. Naturally infected clams were exposed to different water-quality treatments and monitoring in the laboratory. In a high-temperature treatment (26 C vs. control 18 C), the mean abundance of the parasite decreased to near zero in 7 days. Because only 2 clams of 72 died in this treatment during the 14-day experiment, the survival of the host was not seriously at risk at the high temperature. 'Low oxygen, no water change' (18 C) was the second most effective treatment, followed by a 'low-oxygen, water-flow' (18 C) treatment. At the end of the experiment, the mean parasite abundance was significantly lower in all the treatments than in the control clams (18 C). A few P. rylovi individuals abandoned the host at 26 C but died in a couple of days outside the host. However, the parasites lived on average (+/-SE) 12.7 +/- 0.9 days outside the clam, and were also shown to be capable of infecting another uninfected host individual, at 18 C. The results of the present study suggest that high temperature provides an effective, ecologically sustainable method to manipulate the intensity of P. rylovi infection. PMID- 14740907 TI - New species of myxosporean (Myxozoa: Myxosporea) parasites of Ceratomyxa from fishes of Peter the Great Bay (Japan Sea). AB - Four new species of Ceratomyxa were found during parasitological studies of fish caught in shallow areas of Peter the Great Bay, Russia. Two of them (C. aspera n. sp. and C. durusa n. sp.) were found in the gall bladders of the flounders Limanda aspera and L. herzensteini. The third species (C. azonusi n. sp.) infected the gall bladder of the greenling Pleurogrammus azonus, and the fourth (C. lianoides n. sp.) was found in the gall bladder of Stichaeus grigorjewi. Ceratomyxa spp. have not been previously described from P. azonus or S. grigorjewi. PMID- 14740908 TI - A new species of Mathevotaenia (Cestoda: Anoplocephalidae) and other tapeworms from marsupials in Argentina. AB - Cestodes are reported from Didelphis albiventris Lund, 1840 and Micoureus cinereus Temminck, 1824 (Marsupialia: Didelphidae) in Argentina. These include a new species of Mathevotaenia Akhumyan, 1946 (Cestoda: Anoplocephalata) as well as M. bivittata (Janicki, 1904) and an unknown hymenolepidid cestode. Mathevotaenia argentinensis n. sp. is characterized by a relatively narrow strobila, 18-37 mm in total length and 1.0-1.5 mm in maximum width, 135-163 craspedote proglottids, 19-27 testes, and a muscular genital atrium. This species differs from M. didelphidis (Rudolphi, 1819) in the disposition of the genital ducts between the excretory canals and in the entrance of the vagina into the genital atrium posterior to the cirrus pouch; from M. paraguayae Schmidt and Martin, 1978 in the disposition of the genital ducts, absence of a seminal receptacle, and presence of an armed cirrus; and from M. boliviana Sawada and Harada, 1986 and M. pennsylvanica Chandler and Melvin, 1951 in the presence of an armed cirrus. Linstowiines appear to be the dominant cestodes in New World marsupials, with M. bivittata representing the most prevalent and widely distributed species. The hymenolepidid is the first record of this family in Neotropical marsupials. PMID- 14740909 TI - Strelkovimermis papillosus n. sp. (Nematoda: Mermithidae), a parasite of chironomid (Insecta: Diptera) adults from the headwaters of the Mississippi River in northern Minnesota. AB - On 18 August 2002, chironomid imagoes of Rheotanytarsus sp. emerged from the upper Mississippi River in Minnesota and yielded distinctive mermithid nematodes of a new mermithid species. Strelkovimermis papillosus n. sp. is distinguished from the other 14 species of the genus by the presence of unusually large cephalic papillae encircling the mouth and forming a rosette with the mouth in the center and by the absence of a fixator muscle in the males. Additionally, both sexes have very acute posterior ends, long amphids, and a long stoma. Strelkovimermis is revised by eliminating nondiscriminating parameters and accommodating the 15 known species. Intrageneric characteristics useful in separating species of Strelkovimermis are listed. Intensity of infection and intensity of infection versus sex were determined from 41 hosts. Where known, the hosts and geographical distribution are given for all 15 Strelkovimermis species. PMID- 14740910 TI - Molecular phylogeny and surface morphology of marine aseptate gregarines (Apicomplexa): Selenidium spp. and Lecudina spp. AB - Many aseptate gregarines from marine invertebrate hosts are thought to have retained several plesiomorphic characteristics and are instrumental in understanding the early evolution of intracellular parasitism in apicomplexans and the phylogenetic position of cryptosporidians. We sequenced the small-subunit (SSU) ribosomal RNA genes from 2 archigregarines, Selenidium terebellae and Selenidium vivax, and 2 morphotypes of the marine eugregarine Lecudina polymorpha. We also used scanning electron microscopy to investigate the surface morphology of trophozoites from Lecudina tuzetae, Monocystis agilis, the 2 species of Selenidium, and the 2 morphotypes of L. polymorpha. The SSU ribosomal DNA sequences from S. vivax and L. polymorpha had long branch lengths characteristic of other gregarine sequences. However, the sequence from S. terebellae was not exceptionally divergent and consistently emerged as 1 of the earliest 'true' gregarines in phylogenetic analyses. Statistical support for the sister relationship between Cryptosporidium spp. and gregarines was significantly bolstered in analyses including the sequence from S. terebellae but excluding the longest branches in the alignment. Eugregarines formed a monophyletic group with the neogregarine Ophryocystis, suggesting that trophozoites with elaborate cortex folds and gliding motility evolved only once. The trophozoites from the 2 species of Selenidium shared novel transverse striations but differed from one another in overall cell morphologies and writhing behavior. PMID- 14740911 TI - Molecular phylogeny of Haematoloechus Looss, 1899 (Digenea: Plagiorchiidae), with emphasis on North American species. AB - Phylogenetic hypothesis of 23 populations corresponding to 18 species of the digenean Haematoloechus from America, Europe, and Africa, based on ribosomal DNA 28S partial sequences (approximately 890 bp), is presented. Genetic divergence between the in-group and the out-groups ranged from 9.7 to 14.5% and within the in-group, from 0.9 to 12.2%. Eight most parsimonious trees 569 steps long were obtained, with a consistency index of 72%. Groups in the tree are not congruent with those in previous classification schemes of species in the genus, based on a small number of morphological characters. For this subset of Haematoloechus species, plesiomorphic hosts are species of Rana, with 2 colonizations to other amphibian groups. African species appear to have diverged after the separation of Gondwana and Laurasia. Therefore, South American species should appear as the closest relatives of African species when included in the analysis. The evidence presented suggests an ancestral wide distribution of North American representatives of the group, followed by successive contraction, amplification, and fragmentation of ranges and speciation events as a result of the intense volcanic activity in the central part of Mexico since the late Tertiary, the drying climate of western and central United States and northwestern Mexico from the early Eocene to the Pleistocene, and the glaciation during the Pleistocene. PMID- 14740912 TI - Taxonomic status of Choanoscolex lamothei Garcia-Prieto, 1990 (Cestoda: Proteocephalidea) using morphological and molecular evidence. AB - The taxonomic status of Choanoscolex lamothei, a proteocephalidean tapeworm described from the catfish Ictalurus meridionalis (= I. furcatus) from Oaxaca, Mexico, was evaluated on the basis of morphological examination of types and freshly collected specimens from the type host and partial sequences of the large subunit (28S) ribosomal DNA. This study revealed that C. lamothei is a species of Megathylacoides (Proteocephalidae: Corallobothriinae) because of the medullary position of the genitalia (entirely cortical in Choanoscolex), the presence of semispherical sphincters on the suckers, a follicular ovary, and the alternating position of the vagina. Megathylacoides lamothei n. comb. differs from congeneric species in lacking an apical organ and in the number (130-208) of testes. Molecular data confirm the position of M. lamothei within Megathylacoides, which contains species parasitizing ictalurid fishes in North America. PMID- 14740913 TI - Observations of Breinlia booliati in a new host, Rattus rattus jalorensis, from Kuantan, state of Pahang, Malaysia. AB - Breinlia booliati Singh and Ho, 1973 is described from the Malaysian wood rat, Rattus rattus jalorensis Bonhote. The parasites presented here were originally discovered in 1955 in Kuantan, Malaysia, but were not classified until now. On the basis of morphological observations of anatomical structures and comparisons with other species of Breinlia, it was determined that the parasites were B. booliati. The parasites discussed here show slight deviation from B. booliati, but they do not warrant a new species classification. There is some variation in anatomical measurements, the number of male caudal papillae, and the morphology of the microfilariae. Breinlia booliati from a new host is described in this article, with a brief discussion on Rattus species that are hosts of B. booliati and vectors that transmit the parasite. The occurrence of B. booliati in R. r. jalorensis represents the first report of the parasite in this host. PMID- 14740914 TI - Development of an antibody-based capture enzyme-linked immunosorbent assay for detecting Echinostoma caproni (Trematoda) in experimentally infected rats: kinetics of coproantigen excretion. AB - The present study reports on the development of a coproantigen capture enzyme linked immunosorbent assay (ELISA) for detecting Echinostoma caproni in experimentally infected rats. The capture ELISA was based on polyclonal rabbit antibodies that recognize excretory-secretory (ES) antigens. The detection limit of pure ES was 3 ng/ml in sample buffer and 60 ng/ml in fecal samples. The test was evaluated using a follow-up of 10 rats experimentally infected with 100 metacercariae of E. caproni, and the results were compared with those of other diagnostic methods such as parasitological examination and antibody titers determined by indirect ELISA. Coproantigens were detected in all the infected rats from the first day postinfection (DPI). The period of maximal coproantigen excretion was between 7 and 21 DPI. The values remained positive until 49-56 DPI, coinciding with the disappearance of the eggs in the stool samples of the infected rats. The kinetics of coproantigen detection were correlated with those of egg output. The present assay provides an alternative tool for the diagnosis of the echinostome infections. The proposed capture ELISA makes possible an earlier diagnosis than that provided by parasitological examination and indirect ELISA and also allows for the differentiation of past and current infections. Our results show that this assay can also be used to monitor the course of echinostome infections. PMID- 14740915 TI - Spurious genotypes in female nematodes resulting from contamination with male DNA. AB - Females of many invertebrates contain stored sperm or fertilized eggs or both, causing potential genotyping errors. We investigated errors caused by male DNA contamination by amplifying 5 microsatellites in DNA isolated from various tissue types in the nematode Ascaris lumbricoides. We observed additional alleles in 30/135 uterus-derived samples when compared with muscle controls, resulting in 20/135 (15%) incorrect genotypes and an underestimation of inbreeding. In contrast, we observed additional alleles in only 5/143 ovary-derived samples, resulting in 4/143 (3%) incorrect genotypes and no significant influence on inbreeding estimates. Because uterus constitutes approximately 17% of a female's organ weight, a substantial proportion of samples isolated from female tissue may contain male-derived DNA. Male contamination is easily avoided when using large nematodes such as A. lumbricoides. However, we urge caution for studies using DNA isolated from small invertebrates that store sperm or fertilized eggs or both. PMID- 14740916 TI - Lack of detectable shedding of Cryptosporidium parvum oocysts by periparturient dairy cattle. AB - We examined whether periparturient dairy cattle shed Cryptosporidium parvum oocysts within 12 hr of calving on 3 commercial dairy farms endemic for calfhood cryptosporidiosis. Using a diagnostic method that can detect as few as 1 oocyst per gram of feces, we found no evidence of C. parvum oocysts in 86 fecal samples collected within 12 hr of calving from 43 dairy cows. PMID- 14740917 TI - Absence of interferon-gamma-inducible gene IGTP does not significantly alter the development of chagasic cardiomyopathy in mice infected with Trypanosoma cruzi (Brazil strain). AB - Interferon-gamma (IFN-gamma) contributes to host resistance during acute infection with Trypanosoma cruzi, the causative agent of Chagas' disease. Inducibly expressed guanosine triphosphatase (IGTP), a 48-kDa guanosine triphosphatase (GTPase), is a member of a family of GTPase proteins inducibly expressed by IFN-gamma. The expression pattern of IGTP suggests that it may mediate IFN-gamma-induced responses in a variety of cell types. IGTP has been demonstrated to be important for control of Toxoplasma gondii infection but not for resistance against Listeria monocytogenes. We evaluated the role of IGTP in development of chronic chagasic cardiomyopathy in IGTP null mice and C57X129sv (wild type [WT]) mice infected with the Brazil strain for 6 mo. There was no significant difference in parasitemia or cardiac histopathology between null and WT mice. Right ventricular remodeling was observed in infected IGTP null mice, suggesting that IGTP does not significantly alter the course of T. cruzi infection. PMID- 14740918 TI - Seroepidemiology of Toxoplasma gondii infection among Chinese aboriginal and Han people residing in mountainous areas of northern Thailand. AB - A seroepidemiological survey of Toxoplasma gondii infection among Chinese refugees, including Akka and Yau aborigines and Han people living in mountainous areas at elevations of 1,100-1,400 m in Chiang-Rai Province of northern Thailand, was conducted during January 2003 using the latex agglutination test. The overall seroprevalence of T. gondii infection was 9.1% for Akka aborigines, 37.9% for Yau aborigines, and 7.9% for Han people, respectively. No significant gender difference in seroprevalence was found among any of the groups (P > 0.05). The results of a multiple logistic regression analysis for Yau aborigines and Han people showed that the older the age, the higher the odds ratios (OR) of being seropositive (OR = 3.0, 95% confidence interval [CI] = 0.5 to 16.9, P < 0.001 and OR = 1.5, 95% CI = 0.3 to 8.0, P = 0.06 for the elderly group vs. the child group for the Yau aborigines and Han people, respectively). In contrast, the OR was lower among older Akka aboriginal populations (OR = 0.1, 95% CI = 0.0 to 0.4, P < 0.001). Ethnically, Yau aboriginal populations had a significantly higher seroprevalence than did the Akka aborigines and Han people (P < 0.001). PMID- 14740919 TI - The antiparasitic actions of plant jasmonates. AB - Jasmonates are a group of small lipids produced in plants, which function as plant stress hormones. We have previously shown that jasmonates can exert significant cytotoxic effects upon human cancer cells. The purpose of the present study was to determine the effects of jasmonates on parasites. To that end, we chose 2 major human blood parasites, Plasmodium falciparum, a unicellular parasite, and Schistosoma mansoni, a multicellular helminth parasite, and studied the effects of jasmonates on these parasites in vitro. We found that jasmonates are cytotoxic toward both parasites, with P. falciparum being the more susceptible. Jasmonates did not cause any damage to control human erythrocytes at the maximum concentration used in the experiments. This is the first study demonstrating the antiparasitic potential of plant-derived jasmonates. PMID- 14740920 TI - Surface ultrastructure of the advanced third-stage larvae of Gnathostoma nipponicum. AB - The surface ultrastructure of advanced third-stage larvae (AL3) of Gnathostoma nipponicum was studied using scanning electron microscopy. The larvae were recovered from the grass snake Rhabdophis tigrina in the Republic of Korea. Parasites had a globular head bulb with a pair of lips at the anterior end and 2 labial papillae and an amphid on each lip. The head bulb was characteristically armed with 3 transverse rows of hooklets, averaging 36, 38, and 43 in number, increasing posteriorly. A total of 213-232 minute unidentate cuticular spines were present along the entire length of the larvae, forming the transverse striations. Two pairs of cervical papillae were located between the 8th and 12th transverse striations, and a pair of body papillae was seen laterally on the posterior third of the body. A pair of caudal phasmids was recognized near the posterior extremity. The surface ultrastructure of AL3 of G. nipponicum is unique compared with that of other species. PMID- 14740921 TI - Miracidia of an Egyptian strain of Schistosoma mansoni differentiate between sympatric snail species. AB - The host-finding behavior of miracidia of 2 strains of Schistosoma mansoni from Egypt and Brazil was studied by recording their responses to snail-conditioned water (SCW) from the Egyptian sympatric snails, Biomphalaria alexandrina, Physa acuta, Lymnaea cailliudi, and Balinus truncatus, as well as from Biomphalaria arabica and Biomphalaria glabrata. Miracidia of the Egyptian strain significantly preferred SCW from their compatible hosts B. alexandrina and B. arabica and showed no or a weak response to SCW from the other sympatric species, whereas miracidia of the Brazilian strain did not differentiate between SCW from different snail species. PMID- 14740922 TI - Detection of genetic variation in Taenia solium. AB - Genetic variability among Taenia solium isolates was studied in 160 cysticerci from 6 pigs, 4 from Mexico, 1 from Honduras, and 1 from Tanzania. Random amplified polymorphic DNA (RAPD) analysis performed with 4 commercial primers showed 88% polymorphic loci and an average heterozygosity of 0.077; however, several alleles were fixed within each isolate. Linkage disequilibrium analysis indicated that 3 of the 6 isolates had a random association of alleles, whereas the other 3 had a clonal structure. These results suggest the existence of local lineages in T. solium, with events of genetic recombination within them. PMID- 14740923 TI - Identification of assemblage A Giardia in white-tailed deer. AB - Fecal samples were collected from hunter-killed white-tailed deer (Odocoileus virginianus) during a managed hunt in a central Maryland county. Fecal samples were cleaned of debris and concentrated by CsCl density gradient centrifugation and stained with MerIFluor reagents. Stained samples were examined by fluorescent microscopy for the presence of Giardia sp. cysts. One of 26 samples was found to be positive for Giardia sp. Polymerase chain reaction amplification using primers directed to the beta-giardin and TPI genes identified the same sample as the only positive one. Sequencing of the beta-giardin and TPI genes revealed that the Giardia sp. belonged to assemblage A, a genotype infectious for humans and also reported in a small percentage of cattle. This is the first report of assemblage A Giardia sp. in deer and suggests that deer could be a potential source of infectious cysts for humans and cattle. PMID- 14740924 TI - Bacteriolytic activity of selected vertebrate sera for Borrelia burgdorferi sensu stricto and Borrelia bissettii. AB - An in vitro assay to evaluate the bacteriolytic activity of the complement pathway was applied to 2 strains of Borrelia bissettii, CO501 and DN127, and compared with that of B. burgdorferi sensu stricto B31. Sera from mule deer (Odocoileus hemionus) and the Western Fence lizard (Sceloporus occidentalis) were completely borreliacidal for B. burgdorferi and for both strains of B. bissettii. Serum from Bobwhite quail (Colinus virginianus) was nonlytic for B. burgdorferi and partially lytic for B. bissettii strains, CO-501 and DN127. Serum from a New Zealand White rabbit (Oryctolagus cuniculus) was partially lytic for all 3 strains of Borrelia, whereas serum from white-footed mice (Peromyscus leucopus) were nonlytic for all 3 Borrelia strains. The spectrum of complement sensitivity of B. bissettii appears to be similar to that of European B. afzelii in that tested rodent serum is not lytic to these 2 genospecies. Interestingly, both B. bissettii and B. afzelii have been found to be closely associated with rodents. Complement sensitivity demonstrated in these experiments may suggest and possibly predict specific reservoir-host associations. PMID- 14740925 TI - Hepatitis associated with a Sarcocystis canis-like protozoan in a Hawaiian monk seal (Monachus schauinslandi). AB - A Hawaiian monk seal (Monachus schauinslandi) died in captivity at the National Marine Fisheries Service, Kewalo Basin Facility in Honolulu, Hawaii. The animal was icteric, and the liver was friable. Microscopic lesions were detected in the colon and liver. Colonic lesions included multifocal, necrohemorrhagic colitis associated with gram-negative bacilli. The liver lesions included random hepatic necrosis and cholestasis. Asexual stages of a Sarcocystis canis-like apicomplexan were detected in hepatocytes. The parasite divided by endopolygeny. Merozoites occasionally formed rosettes around a central residual body. Ultrastructurally, merozoites lacked rhoptries. This is the first report of S. canis infection in M. schauinslandi, which is an endangered pinniped in U.S. waters. PMID- 14740926 TI - On polymerase chain reaction tests for estimating prevalence of malaria in birds. PMID- 14740927 TI - Ultrastructural study of muscle in vocal fold: postmortem damage observations. AB - Various authors have published results related to the ultrastructure of vocal folds in specific areas proceeding from human cadavers. Nevertheless, starting from the premise that a fundamental principle of sampling and the samples should be a true representative of the whole, the authors decided to examine vocal folds from human cadavers and compare them to normal vocal folds from the guinea pig (Cavia porcellus). The findings of these authors demonstrated clearly that the conclusion of Rhodin (1954) is correct, that a biological sample must be preserved immediately after blood circulation ceases, and that the intermediary time between biopsy and fixation cannot be more than 3 minutes. Cells fixed a few hours after death appeared disaggregated, and many of their endocellular components were profoundly altered. The results obtained by the authors suggests that ultrastructural studies with cadaverous material may lead to serious risks or doubts about the accuracy of the results and consequently result in dubious interpretations. PMID- 14740928 TI - Determination of velum opening for French nasal vowels by magnetic resonance imaging. AB - MRI techniques have been used to describe velum opening of French vowels. Data based on 18 joined axial slices of 4 mm thickness were recorded with four subjects. Differences in velum opening are calculated from areas measured in the tract between the lowered velum and the back pharynx wall. Results show that for all subjects, the back vowel [symbol: see text] has the smallest opening, while some variations are observed for the other vowels. PMID- 14740929 TI - Influence of glottal closure configuration on vocal efficacy in young normal speaking women. AB - Posterior closure insufficiency of the glottis is often mentioned in connection with permanent voice disorders. Recently published studies have revealed that an incomplete closure of the glottis can be found also in normal-speaking voices, especially in women. However, the effect of glottal closure configuration on vocal efficacy is not sufficiently clarified. The purpose of this study was to determine the effect of glottal closure configuration on singing and speaking voice characteristics. Overall, 520 young female normal-speaking subjects were examined by videostroboscopy for different phonation conditions in the combination of soft, loud, low, and/or high phonation and by voice range profile measurements. According to the videostroboscopic analysis, the subjects were subdivided into four groups: complete closure of the vocal folds already in soft phonation (group 1), closure of the vocal fold with increasing intensity (group 2), persistent closure insufficiencies despite increasing intensity (group 3), and hourglass-shaped closure in subjects with vocal nodules (group 4). Subjects in which the glottal closure could not be evaluated sufficiently were subclassified into group 5 (missing values). Selected criteria of the singing and speaking voice were evaluated and statistically processed according to the mentioned subclassification. Group 1 reached significantly the highest sound pressure levels (SPLmax) for the singing voice as well as for the shouting voice. Group 3 showed a limited capacity to increase the intensity of the singing and speaking voice. The results gathered in this study objectify the relationship of insufficient glottal closure and reduced vocal capabilities. As long as no conclusive data on long-term consequences of insufficient glottal closure are available, a prophylactic improvement of the laryngeal situation especially in female professional voice users by voice therapy should be recommended. PMID- 14740930 TI - Just noticeable differences of open quotient and asymmetry coefficient in singing voice. AB - This study aims to explore the perceptual relevance of the variations of glottal flow parameters and to what extent a small variation can be detected. Just Noticeable Differences (JNDs) have been measured for three values of open quotient (0.4, 0.6, and 0.8) and two values of asymmetry coefficient (2/3 and 0.8), and the effect of changes of vowel, pitch, vibrato, and amplitude parameters has been tested. Two main groups of subjects have been analyzed: a group of 20 untrained subjects and a group of 10 trained subjects. The results show that the JND for open quotient is highly dependent on the target value: an increase of the JND is noticed when the open quotient target value is increased. The relative JND is constant: deltaOq/Oq = 14% for the untrained and 10% for the trained. In the same way, the JND for asymmetry coefficient is also slightly dependent on the target value--an increase of the asymmetry coefficient value leads to a decrease of the JND. The results show that there is no effect from the selected vowel or frequency (two values have been tested), but that the addition of a vibrato has a small effect on the JND of open quotient. The choice of an amplitude parameter also has a great effect on the JND of open quotient. PMID- 14740931 TI - Change in singing voice production, objectively measured. AB - Although subglottal pressures in conversational speech are relatively easily measured and thus known, the higher values that sometimes occur in singing (especially in tenors) have received little attention in the literature. Still more unusual is the opportunity to measure a large-scale change over decades in the application of pressure in singing production. This study compares measurements of subglottal pressure in a tenor/singing teacher (JS) at two points in his career: in his early thirties, when he was a subject in HS's dissertation study on the efficiency of voice production; and recently, in his fifties, in connection with JS's forthcoming book on the history of the pedagogy of Bel Canto. Although a single case study, its points of special interest include the high values initially measured (up to 100 cm H2O) and the reduction of this figure by more than 50% in the maximal values of the recent measurements. The study compares these values with those of other singers in the same laboratory (both with esophageal balloon and directly, with a catheter passed through the glottis) and in the literature, as well as discusses in detail the problems pertaining to the measurement (repeatability, correcting for lung volume, etc.). As a sophisticated subject, JS makes some pertinent observations about the changes in his use of subglottal pressure. PMID- 14740932 TI - The use of an auditory model in predicting perceptual ratings of breathy voice quality. AB - Despite much research, the relationship between vocal acoustic signals and perceived voice quality is not well understood. The present study used an auditory model proposed by Moore et al to study how changes in the acoustic spectrum may relate to changes in perceptual ratings of breathiness. Perceptual ratings of breathiness were obtained using a multidimensional scaling (MDS) design. The stimulus distances on the dominant MDS dimension were correlated with several commonly used acoustic measures for voice quality. These distances were also compared with measures obtained from the output of the auditory model. Results show that the partial loudness of the harmonic energy obtained with the aspiration noise acting as a masker was the most important predictor of perceptual ratings of breathiness. Results also demonstrate that measures obtained from the auditory spectrum were better predictors of perceptual ratings of breathiness than were commonly used acoustic spectral measures. PMID- 14740933 TI - Laryngeal function and vocal fatigue after prolonged reading in individuals with unilateral vocal fold paralysis. AB - The purpose of the present study was to examine the effect of prolonged loud reading, intended to induce fatigue, on vocal function in adults with unilateral vocal fold paralysis (UVFP). Subjects were 20 adults, 37-60 years old, with UVFP secondary to recurrent laryngeal nerve paralysis. Subjective ratings and instrumental measures of vocal function were obtained before and after reading. Statistical analysis revealed subjects rated their vocal quality and physical effort for voicing more severely following prolonged loud reading, whereas expert raters did not detect a significant perceptual difference in vocal quality. Reading fundamental frequency (F0) was significantly increased following prolonged loud reading, as were mean airflow rates at all pitch conditions. Maximum phonation times for comfort and low pitches significantly decreased during posttests. Multiple regression analyses revealed significant associations between ratings of posttest physical effort and select posttest measures. Interpretation of results indicates the prolonged loud reading task was successful in vocally fatiguing most of the UVFP subjects. Key physiologic correlates of vocal fatigue, in individuals with UVFP, include further reduction of glottic efficiency, resulting in decreased regulation of glottic airflow and a temporary destabilization of speaking fundamental frequency. PMID- 14740934 TI - Effects of vocal training on the acoustic parameters of the singing voice. AB - Vocal training (VT) has, in part, been associated with the distinctions in the physiological, acoustic, and perceptual parameters found in singers' voices versus the voices of nonsingers. This study provides information on the changes in the singing voice as a function of VT over time. Fourteen college voice majors (12 females and 2 males; age range, 17-20 years) were recorded while singing, once a semester, for four consecutive semesters. Acoustic measures included fundamental frequency (F0) and sound pressure level (SPL) of the 10% and 90% levels of the maximum phonational frequency range (MPFR), vibrato pulses per second, vibrato amplitude variation, and the presence of the singer's formant. Results indicated that VT had a significant effect on the MPFR. F0 and SPL of the 90% level of the MPFR and the 90-10% range increased significantly as VT progressed. However, no vibrato or singers' formant differences were detected as a function of training. This longitudinal study not only validates previous cross sectional research, ie, that VT has a significant effect on the singing voice, but also it demonstrates that these effects can be acoustically detected by the fourth semester of college vocal training. PMID- 14740935 TI - Effects of voice therapy on the voice range profiles of dysphonic patients. AB - In a group of chronically dysphonic patients, a voice range profile, or phonetogram, was recorded before and after receiving voice therapy and again 3 months later. The voice range profiles took a wide variety of shapes. Therefore, only measures that did not depend on a smooth contour could be used to describe changes before and after therapy. The main effect of voice therapy was an enlargement on the side of low frequency and low intensity. PMID- 14740936 TI - Perceptual and acoustic evaluation of individuals with laryngopharyngeal reflux pre- and post-treatment. AB - Thirteen individuals with laryngopharyngeal reflux (LPR) were studied pre- and post-treatment. The effect of treatment on perceptual ratings of voice quality and frequency and intensity measures was examined. Relationships between perceptual and acoustic parameters were assessed descriptively. Results showed a small, but significant improvement in the perception of voice quality post treatment. No significant differences were found between pre- and post-treatment means for any of the acoustic measures except harmonics-to-noise ratio (HNR). Descriptive analyses showed some association between perceptual ratings and acoustic measures. Discussion of results focuses on severity of LPR. PMID- 14740937 TI - Tracking outcomes after phonosurgery for sulcus vocalis: a case report. AB - Outcomes data after a surgical or behavioral intervention should be tracked until stability is reached. Often it is unclear how long patients should be followed and at what point an outcome can be considered stable. These issues have implications for treatment decision making, efficacy measurement, and the design of research studies. Vocal function data were collected 24 hours before and at 1, 6, and 12 months after phonosurgery for sulcus vocalis. One data series was collected daily during the first month after surgery, providing a unique opportunity to study voice changes in the immediate postoperative period. The different vocal function indices (acoustic, perceptual, videostroboscopic, aerodynamic, psychosocial) demonstrated a general pattern of improvement after intervention; however, they appeared to reach stability at different times. This report reinforces the value of following patients until complete outcome stability. PMID- 14740938 TI - Determining the etiology of mild vocal fold hypomobility. AB - The prevalence of mild vocal fold hypomobility is unknown. In a study by Heman Ackah et al, vocal fold hypomobility in a population of singing teachers was found to be associated more frequently with vocal complaints than was the presence of vocal fold masses. The etiology of mild vocal fold hypomobility has not been previously explored. In the present study, a retrospective chart review was performed of 134 patients who presented to a tertiary laryngology referral center over a 6-month period for evaluation of vocal complaints. Of the 134 patients, 61 (46%) were found to have mild vocal referring otolaryngologist. Imaging studies and laboratory tests to evaluate for structural, metabolic, and infectious causes of the decreased mobility had been ordered. Forty-nine patients completed the work-up. Of these, 41 out of 49 (84%) were found to have imaging or laboratory findings that could explain the hypomobility. Thyroid abnormalities were found to be associated with vocal fold hypomobility in 21 out of 49 (43%) of those with a complete evaluation. Other causes of vocal fold hypomobility included idiopathic (8 of 49, 16%), viral neuritis (5 of 49, 10%), central nervous system abnormality (4 of 49, 8%), neural tumor (3 of 49, 6%), joint dysfunction (3 of 49, 6%), iatrogenic nerve injury (2 of 49, 4%), myopathy (2 of 49, 4%), and noniatrogenic traumatic nerve injury (1 of 49, 2%), This study shows that unilateral vocal fold hypomobility often is associated with a physiologic process, and a complete investigation to determine the etiology is warranted in all cases. PMID- 14740939 TI - Early results of transcutaneous injection laryngoplasty with micronized acellular dermis versus type-I thyroplasty for glottic incompetence dysphonia due to unilateral vocal fold paralysis. AB - Medialization thyroplasty (type I) has become the gold standard to improve glottic closure due to unilateral vocal fold paralysis. A newer injection method utilizing homologous collagen from cadaveric human tissue has been described as an attractive alternative as no donor site is required, there is a very low risk of hypersensitivity, and the intact, acellular collagen fibers may suffer a reduced long-term reabsorption rate. Preliminary results on eight patients comparing presurgical and postsurgical parameters (perceptual, stroboscopic, acoustic, and aerodynamic) revealed comparable results when compared with a control group of individuals, age- and sex-matched, that had undergone standard medialization thyroplasty (type I). Further study is needed to assess the long term results with this minimally invasive method of vocal fold medialization. PMID- 14740940 TI - Comparison between thyroplasty type I and arytenoid rotation: a study of vocal fold vibration using excised human larynges. AB - The purpose of this paper was to compare the vibration of the vocal fold submitted to Isshiki thyroplasty type I (TPI) to that of the contralateral one adducted by the arytenoid rotation (AR) technique. The vocal folds of ten human fresh excised larynges were medialized by TPI on one side and by rotation of the arytenoid on the contralateral side. Laryngeal vibration was artificially produced and was recorded by videostroboscopy. The images were subjectively and objectively analyzed. Subjective analysis included periodicity of vibratory cycles, features of the mucosal wave present on the TPI side, amplitude of vibration, and profile of free border of each vocal fold during the opening phase. Objective analyses were carried out on frame-by-frame digitalized images to determine amplitudes of vibrations and phase differences between the folds in three glottic regions (anterior, middle, and posterior). Subjective analysis revealed regular periodicity in 100% of the larynges, a decrease in the mucosal wave on the TPI side in 70%, reduction in amplitude in 30%, and a sigmoid profile of the free border on the TPI side in 80%. Objective analysis showed mean amplitude in the posterior glottic region on the TPI side significantly larger than that on the arytenoids rotation side and phase asymmetry in 90% of the larynges. PMID- 14740941 TI - Ethical considerations in newer reproductive technologies. AB - Ethics is an essential dimension of newer reproductive technologies. In this rapidly evolving field, there is a need for an ethical framework to guide both clinical practice and research. In this article, we provide such a framework that incorporates ethical principles, professional virtues, and the concept of the fetus as a patient. We then apply this framework to an important current clinical issue: the number of embryos to be transferred during in vitro fertilization; in addition to an emerging research issue: gene transfer research on in vitro embryos. PMID- 14740942 TI - Patient refusal of treatment in obstetrics. AB - During the later half of the previous century, Clinical Ethics evolved from the iconoclastic and murky realm of philosophical dissertation into a practical and useful discipline to be applied to clinical dilemmas occurring in everyday practice. We have explored a brief exposition of prevailing ethical theories that are most commonly recognized. These include Consequentialism and Principlism, and we have applied them to actual cases as they might occur in clinical practice. It is our hope that this will help us all to facilitate decisions with which we will be comfortable and which will be compatible with the beliefs and values of our patients. PMID- 14740943 TI - Ethical issues in fetal surgery. AB - Fetal surgery involves unique ethical issues because the interests of fetus and mother may conflict but, regardless, they are inextricably interconnected. Controversial questions currently include what kinds of surgery should be permitted, who should decide, and whether this surgery should be restricted to specialized centers. Clinical questions include the degree to which mothers should have decision-making authority, the extent, if any, to which mothers should be protected from pressure from family members, and whether physicians should be non-directive. This article discusses these questions and their answers' competing rationales. It also presents recent data from attachment studies of mothers and infants and from neuroscience, which suggest that mothers and infants are more interdependent physiologically and psychologically than has been understood. The paper describes how these findings may apply to the mother and fetus long before it is born. The major implication of this analysis is that mothers should, perhaps, have greater decision making authority. PMID- 14740944 TI - Ethical considerations in the management of infants born at extremely low gestational age. AB - With ongoing improvements in technology and the understanding of neonatal physiology, there has been increasing debate regarding the gestational age and birth weight limits of an infants' capability of sustaining life outside the womb and how this is to be determined. The objective of this review was to address this issue with an analysis of current data (following the introduction of surfactant therapy in 1990) from published studies of survival in extremely low gestational age infants. We found that survival was possible at 22 completed weeks of gestation but only in < 4% of live births reported. Survival increased from 21% at 23 weeks gestational age to 46% at 24 weeks gestational age. Historically, despite continual advances in neonatology, the mortality at 22 weeks has not improved over the past three decades. Combining the data from studies on survival with evidence from developmental biology, we believe that it is not worthwhile to pursue aggressive support of infants born at < 23 weeks gestational age. Given the complicated issues related to morbidity and mortality in infants born at 22 to 25 weeks gestational age and the ethical implications of the available evidence, we propose the need for a consensus derived framework to help in decision-making. PMID- 14740945 TI - Some are old, some are new: life and death in the ICU. AB - We compared 560 adults hospitalized in our Medical Intensive Care Unit (MICU) to 245 ventilated babies hospitalized in our Neonatal ICU (NICU). Both ICUs had comparable mortality rates--roughly 1 patient in 5 died. The average length of hospitalization for nonsurvivors versus survivors was disproportionately short for NICU babies (13d v 33d) and long for MICU adults (15d v 12d). This phenomenon resulted in a redistribution of ICU bed-days and resources in favor of survivors for NICU babies (approximately 9 of every 10 NICU beds were devoted to babies who survived), and nonsurvivors for MICU adults (roughly 1 MICU bed in 2). Both ICUs had comparable percentages of patients predicted to die--roughly 1 patient in 3. The predictive power of an intuition of die was comparable--and not all that great. Almost one third of patients in both ICUs with a single prediction of "die in hospital" survived to be discharged. However, the likelihood of finding a neurologically normal NICU survivor after a prediction of "die" was only 5 in 100. To the extent that informed decisions can be made with 95% certainty, we may have found a foothold on the slippery ethical slope of benefit/burden calculations in the NICU. Unfortunately, we have no comparable data for MICU survivors. PMID- 14740946 TI - The ethics of withholding/withdrawing nutrition in the newborn. AB - The provision of nutrition and hydration to newborn infants is considered fundamental care. For premature and critically ill newborns, similar considerations generally hold true. Nutrition may be provided for these infants using assisted measures such as parenteral nutrition or tube feedings. However, for some newborn infants the provision of medically assisted nutrition may be a more complicated issue. In particular, the goals of nutrition need to be clearly elaborated for newborns with lethal conditions or for whom appropriately administered intensive care is unsuccessful in sustaining life. These infants may benefit from palliative measures of care and a limitation or withdrawal of burdensome or nonbeneficial interventions. This article explores issues pertinent to deciding and communicating the appropriate withdrawal of medically assisted nutrition and implementing palliative comfort measures. PMID- 14740947 TI - Deciding for neonates: whose authority, whose interests? AB - Two principles are commonly perceived as being central to decision-making for critically ill newborns: the patient's best interest standard, and the authority of the parents to speak for the newborn as the surrogate decision-makers. In this essay, these 2 principles are examined in the context of a particular setting, that of a critically ill newborn with a mother in early adolescence. Alternatives to the patient's best interest standard are explored, including consideration of other interests in addition to those of the patient, such as the interests of the young mother and other family members. Also, we discuss a model of shared decision-making, wherein "parental authority" may be shared between the young adolescent mother and an adult relative, such as her own mother. PMID- 14740948 TI - Resolving disputes between clinicians and family about "futility" of treatment. AB - Family resistance to withdrawal of life support from children presents difficult issues of clinical practice and of principle. Legal recognition of unilateral physician authority for withdrawal on grounds of clinical "futility"-even in the most extreme circumstance of brain death-creates inappropriate incentives for clinicians' avoidance of prolonged, emotionally taxing interactions likely to persuade parents to accept the reality of their child's impending or actual death. Although unilateral physician action withdrawing support may sometimes be necessary in response to intractable family resistance, clinicians should nonetheless always understand this course not only as a clinical failure in dealing with families but also as unjustified in principle. PMID- 14740949 TI - Biologic properties of dengue viruses following serial passage in primary dog kidney cells: studies at the University of Hawaii. AB - Serial passage at low dilution of seven different wild-type dengue (DEN) viruses into primary dog kidney (PDK) cell cultures placed selective pressure that resulted in the following changes from parental phenotype: smaller plaques in LLC MK2 cells, absent plaque formation in green monkey kidney cells, lack of a cytopathic effect in LLC-MK2 cells, shut-off of virus replication at high temperatures (temperature sensitivity), reduced virulence for rhesus monkeys manifested by reduced or absent viremia and/or absence of a secondary-type antibody response following homotypic challenge, and progressive increase in the mean day of death following intracerebral inoculation of sucking mice. Two DEN-1 strains showed most of these changes by the 15th PDK passage. Only one of two DEN 2 strains studied was carried to the 50th PDK passage at the University of Hawaii. For the latter strain, both the temperature of viral replicative shutoff and mouse neurovirulence were reduced. Three DEN-4 strains showed similar late passage biologic marker changes. The observations made, although not exhaustive, provide laboratory correlates for virus strains that have shown reduced virulence but retained immunogenicity in humans. Candidate human vaccines have been prepared from five of the studied strains: DEN-1 (16007) at PDK 13, DEN-2 (16681 and S-16803) at PDK 50 or above, and DEN-4 (1036 and 341750) at PDK 48 and 20, respectively. PMID- 14740950 TI - Modification of dengue virus strains by passage in primary dog kidney cells: preparation of candidate vaccines and immunization of monkeys. AB - Dengue (DENV) virus strains for each of the four DENV serotypes were modified by passage in primary dog kidney (PDK) cell cultures with final manufacture of vaccine lots in fetal rhesus monkey diploid cell cultures. "Strain sets" consisting of serially-passaged DENV were inoculated in rhesus monkeys along with unmodified parent viruses for each strain. Vaccine candidates were compared with unmodified parent viruses by measuring viremia and immune responses. All except one DENV-1 strain demonstrated reduced infection in monkeys after PDK cell passage. A DENV-3 strain lost all monkey infectivity after PDK cell passage. Twelve vaccine candidates were selected for Phase 1 human trials through this selection process. PMID- 14740951 TI - Phase 1 studies of Walter Reed Army Institute of Research candidate attenuated dengue vaccines: selection of safe and immunogenic monovalent vaccines. AB - We describe the results of initial safety testing of 10 live-attenuated dengue virus (DENV) vaccine candidates modified by serial passage in primary dog kidney (PDK) cells at the Walter Reed Army Institute of Research. The Phase 1 studies, conducted in 65 volunteers, were designed to select an attenuated vaccine candidate for each DENV serotype. No recipient of the DENV candidate vaccines sustained serious injury or required treatment. Three vaccine candidates were associated with transient idiosyncratic reactions in one volunteer each, resulting in their withdrawal from further clinical development. Increasing PDK cell passage of DENV-1, DENV-2, and DENV-3 candidate vaccines increased attenuation for volunteers, yet also decreased infectivity and immunogenicity. This effect was less clear for DENV-4 candidate vaccines following 15 and 20 PDK cell passages. Only one passage level each of the tested DENV-2, -3, and -4 vaccine candidates was judged acceptably reactogenic and suitable for expanded clinical study. Subsequent studies with more recipients will further establish safety and immunogenicity of the four selected vaccine candidates: DENV-1 45AZ5 PDK 20, DENV-2 S16803 PDK 50, DENV-3 CH53489 PDK 20, and DENV-4 341750 PDK 20. PMID- 14740952 TI - Vaccination of human volunteers with monovalent and tetravalent live-attenuated dengue vaccine candidates. AB - Four serotypes of monovalent live attenuated dengue virus vaccine candidates were tested for reactogenicity and immunogenicity in 49 flavivirus non-immune adult human volunteers. The four monovalent candidates were then combined into a tetravalent formulation and given to another 10 volunteers. Neutralizing antibody seroconversion rates after a single-dose monovalent vaccination ranged from 53% to 100%. Solicited reactogenicity was scored by each volunteer. A composite index, the Reactogenicity Index, was derived by these self-reported scores. Reactogenicity differed among the four serotype candidates with serotype-1 associated with the most vaccine related side effects. A second dose of monovalent vaccines at either 30 days or 90 days was much less reactogenic but did not significantly increase seroconversion rates. Seroconversion rates in the 10 volunteers who received a single dose of tetravalent vaccine ranged from 30% to 70% among the four serotypes. Similar to the monovalent vaccines, a second dose of the tetravalent vaccine at one month was less reactogenic and did not increase seroconversion. A third dose of the tetravalent vaccine at four months resulted in three of four volunteers with trivalent or tetravalent high-titer neutralizing antibody responses. PMID- 14740953 TI - Atypical antibody responses in dengue vaccine recipients. AB - Eight of 69 (12%) healthy adult volunteers vaccinated with monovalent live attenuated dengue virus (DENV) vaccine candidates had atypical antibody responses, with depressed IgM:IgG antibody ratios and induction of high-titer hemagglutination-inhibiting and neutralizing (NT) antibodies to all four DENV serotypes. These features suggested flavivirus exposure prior to DENV vaccination, yet no volunteer had a history of previous flavivirus infection, flavivirus vaccination, or antibody to flaviviruses evident before DENV vaccination. Moreover, production of antibody to DENV by atypical responders (AR) was not accelerated compared with antibody responses in the 61 flavivirus-naive responders (NR). Further evaluation revealed no differences in sex, age, race, DENV vaccine candidate received, or clinical signs and symptoms following vaccination between AR and NR. However, viremia was delayed at the onset in AR compared with NR. A comparative panel of all AR and five randomly selected NR found flavivirus cross-reactive antibody after vaccination only in AR. Unexpectedly, six of eight AR had NT antibodies to yellow fever virus (YFV) > 1:10 before vaccination while NR had none (P = 0.04). The AR also universally demonstrated YFV NT antibody titers > or = 1:160 after DENV vaccination, whereas four of five NR failed to seroconvert (P = 0.02). Yellow fever virus priming broadens the antibody response to monovalent DENV vaccination. The effect of flavivirus priming on the clinical and immunologic response to tetravalent DENV vaccine remains to be determined. PMID- 14740954 TI - Serotype-specific T(H)1 responses in recipients of two doses of candidate live attenuated dengue virus vaccines. AB - As part of a larger vaccine study, peripheral blood mononuclear cells (PBMC) were collected from volunteers for analysis of vaccine-induced T cell responses. The PBMC were re-stimulated in vitro with live dengue virus and assayed for T(H)1 or T(H)2 memory cell responses. Re-stimulated PBMC from the volunteers predominantly secreted interferon-gamma. Little interleukin-4 (IL-4) or IL-10 secretion was detected, indicating a T(H)1 type of T cell response. The interferon-gamma response was primarily serotype-specific with some serotype cross-reactivity. T cell depletion studies showed that the interferon-gamma was being secreted by CD4+ T lymphocytes and/or by cells other than CD8+ T lymphocytes that were being stimulated by the CD4+ T lymphocytes. CD3+ or CD8+ T cell depletion showed that granzyme B mRNA expression correlated with the presence of CD4+ T lymphocytes. However, depletion of CD4+ T cells after four days of stimulation indicated that the granzyme B mRNA was produced by cells in culture other than lymphocytes. In summary, an antigen-specific T(H)1 type T cell response was seen as a response to vaccination using live attenuated dengue virus. PMID- 14740956 TI - Aesthetics and reconstruction--seeing before cutting. AB - In order to restore a specific form during aesthetic surgery it is self-evident that the surgeon needs to know the form exists. But more than this simple form restoration, the development of aesthetic restoration suggests a specific restoration in the context of the whole form. Developing a disciplined method of visualizing the problem is the first step toward being able to start an aesthetic sense or eye. Careful study of master artists drawings often forces the surgeon to simplify and concentrate not on parts, but parts as they relate to the whole face. Often the artist, as with the successful surgeon, must continually alternate consideration of a specific part with consideration of the whole. This discussion is meant to provide a few relationships and constructs so that the surgeon may start on the journey to development of the aesthetic eye. PMID- 14740955 TI - Phase I trial of 16 formulations of a tetravalent live-attenuated dengue vaccine. AB - Laboratory-attenuated strains of each of the four dengue serotypes previously tested as monovalent vaccines in volunteers were combined and tested for immunogenicity, safety, and reactogenicity in 16 dosage combinations. Tetravalent vaccines made using combinations of high (10(5-6) plaque-forming units [PFU]/dose) or low (10(3.5-4.5) PFU/dose) dosage formulations of each of the four viruses were inoculated in 64 flavivirus non-immune adult volunteers to determine which, if any, formulation raised neutralizing antibodies in at least 75% of volunteers to at least three of four dengue serotypes following one or two inoculations. Such formulations, if safe and sufficiently non-reactogenic, would be considered for an expanded Phase II trial in the future. Formulations 1-15 were each inoculated into three or four volunteers (total = 54) on days 0 and 28. Formulation 16 was tested in 10 volunteers, five volunteers inoculated on days 0 and 30, one volunteer on days 0 and 120, and four volunteers on days 0, 30, and 120. Blood was drawn for serologic assays immediately before and one month after each vaccination, and for viremia assay on day 10 after each vaccination. The 16 formulations were safe, but variably reactogenic after the first vaccination, and nearly non-reactogenic after the second and third vaccinations. Reactogenicity was positively correlated with immunogenicity. Similar proportions of volunteers seroconverted to dengue-1 (69%), dengue-2 (78%), and dengue-3 (69%), but significantly fewer volunteers seroconverted to dengue-4 (38%). The geometric mean 50% plaque reduction neutralization test titers in persons who seroconverted were significantly higher to dengue-1 (1:94) than to dengue-2 (1:15), dengue-3 (1:10), and dengue-4 (1:2). Seven formulations met the serologic criteria required for an expanded trial, and three of these were sufficiently attenuated clinically to justify further testing. PMID- 14740957 TI - Planning the closure. AB - Consistently achieving aesthetically pleasing closures of the skin requires a significant amount of forethought and planning. In addition to understanding the regional anatomy, recognizing and respecting free margins, cosmetic unit junctions, and skin tension lines are all of critical importance. PMID- 14740958 TI - Practical perioral closures. PMID- 14740959 TI - Periocular reconstruction. AB - The periocular area is a complex region with free margins and cosmetic landmarks that are easily distorted during surgical reconstruction. Repair of surgical defects should maintain both periorbital function and aesthetics. In this article we discuss the basic methods of closure (side-to-side closure, flaps, grafts and second intent healing) as they apply to the periocular area and suggest an algorhythmic approach to reconstruction by aesthetic subunit. Pertinent regional anatomy and perioperative management are also reviewed. PMID- 14740960 TI - Closing surgical defects of the external ear. AB - Closing surgical defects of the external ear poses unique challenges because of the convoluted shape and thin tethered skin. Choice of repair is often dictated by the site of the wound. If the defect is central and anterior with intact cartilage, most defects will do well by second intention healing or grafting. If the defect involves the helical rim, reconstruction is often preferred to maintain the normal curvature of the external ear and a helical rim advancement flap with trimming of the central cartilage is often used. Defects of the posterior ear where the skin is more abundant and loose can often be closed side to side. Split earlobes may be repaired by Z-plasty. The full range of repair options should be considered in every case. Because each ear differs in shape and flexibility, creativity is warranted, rewarding both the patient and the surgeon. PMID- 14740961 TI - The examination of several common misconceptions in nasal reconstruction. AB - Because the nose is a very prominent aesthetic feature of the face, wounds located here are particularly challenging from a reconstructive perspective. Poor tissue availability and the potential for introducing anatomic distortion certainly increase surgical complexity. There are common misconceptions regarding nasal reconstruction that may impede the physician's ability to deliver optimal care. Understanding that these commonly held assumptions about nasal wound repair are incorrect should increase the surgeon's nasal reconstructive abilities. The basic principles of reconstructive surgery are not uniquely applicable to the repair of nasal defects; the consequences of surgical misjudgments on the nose are simply magnified in comparison to surgical problems in other less visually prominent locations. To handle nasal surgical wounds with expertise, the dermatologic surgeon should first realize that there are common misconceptions that often hinder effective clinical decision making. By realizing that these incorrect assumptions can reduce success in nasal reconstruction, the surgeon should be able to more appropriately select nasal repair procedures that will accomplish the common goals of any facial reconstructive surgery: to preserve function, to restore appearance, and to minimize surgical risk. PMID- 14740962 TI - Skin grafting. AB - Knowledge of the indications, techniques, donor site considerations, and complications of all types of skin grafting is invaluable for the dermatologic surgeon who performs soft tissue reconstruction on a regular basis. With proper defect assessment, reconstructive planning, and attention to detail pre-, intra-, and postoperatively, optimal cosmetic and functional results using skin grafting techniques can be achieved. PMID- 14740963 TI - Techniques for cutaneous sutured closures: variants and indications. AB - Many different suture techniques are routinely used by cutaneous surgeons for the closure of surgical wounds. This paper reviews several common suture techniques and their associated subtypes. In particular, for each technique, we discuss the method for placement, generally accepted indications, controversies regarding utility, and objective evidence of efficacy. PMID- 14740964 TI - Genetic testing and health insurance: can they coexist? PMID- 14740965 TI - Genetic discrimination arising from cancer risk assessments: a societal dilemma. PMID- 14740966 TI - New fibrinolytic agents for MI: as effective as current agents, but easier to administer. AB - The efficacy and safety of fibrinolytic agents have not dramatically changed since alteplase (Activase), a derivative of tissue plasminogen activator (t-PA), was introduced nearly 2 decades ago. However, newer agents have a longer half life, making them easier to deliver. Fibrinolytic therapy is underused in many patients, especially in those traditionally thought to be at high risk for complications. PMID- 14740967 TI - Should nursing home residents with atrial fibrillation be anticoagulated? AB - Most long-term care residents with atrial fibrillation would be at high risk for embolic stroke based on age and comorbidities according to the criteria presented here. Additionally, they are theoretically excellent candidates for adjusted-dose warfarin treatment for atrial fibrillation. They are accessible for monitoring and tend to have less dietary variability, a controlled medication list, and supervised medication administration. Balancing these features is at least a moderate risk of severe bleeding from anticoagulation based on age, comorbidities, and polypharmacy. However, studies suggest that even those long term care residents identified as ideal candidates for anticoagulation may not receive warfarin. Those residents who do receive warfarin may not be anticoagulated within the therapeutic range much of the time. This treatment pattern may expose older adults with a high stroke risk to dying from a cardioembolic stroke or to acquiring functional deficits that make them more dependent and lower their quality of life. PMID- 14740968 TI - Whole-body CT screening for cancer and coronary disease: does it pass the test? AB - Even though whole-body CT scanning is being marketed directly to patients and they are starting to demand it, does it meet the standards of a good screening test for cancer and coronary artery disease? This article is a step-by-step, disease-specific discussion of the characteristics of a good screening test, and whether whole-body CT scanning meets these standards. PMID- 14740969 TI - How to use statins in patients with chronic liver disease. AB - Clinicians may be concerned about prescribing statins to patients with chronic liver disease, but there is little evidence to suggest that drug-induced liver injury from statins is increased in these patients. Thus, we should prescribe statins for the same indications in patients with chronic liver disease as in patients without, but with closer monitoring. However, patients with acute liver disease (acute viral hepatitis, alcoholic hepatitis) should not take statins until they have recovered. PMID- 14740970 TI - The preoperative evaluation: use the history and physical rather than routine testing. AB - The history and physical examination, rather than routine laboratory, cardiovascular, and pulmonary testing, are the most important components of the preoperative evaluation. The history should include a complete review of systems (especially cardiovascular and pulmonary), medication history, allergies, surgical and anesthetic history, and functional status. PMID- 14740971 TI - Evaluation of the quality of US cancer care: contributions of the ACR Patterns of Care Studies. PMID- 14740972 TI - Conservation therapy in T1-T2 breast cancer: past, current issues, and future challenges and opportunities. AB - PURPOSE: To assess the significance of patient age, race, tumor-related prognostic parameters, status of surgical excision margins, and irradiation boost on incidence of ipsilateral breast relapse, and to review current issues in the management of T1-T2 breast cancer patients with conservation therapy. MATERIALS AND METHODS: Records of 1037 patients with histologically confirmed stage T1 and 308 patients with T2 carcinoma of the breast treated with breast conservation therapy from January 1970 through December 1997 were prospectively registered and evaluated. The mean follow-up for surviving patients was 6.6 years (range, 4-30 years), with a minimum follow up of 4 years for all patients. RESULTS: There were 78 ipsilateral breast relapses (IBRs); the actuarial 10-year incidence of IBR was 7% for T1 and 11% for T2 tumors. In patients 40 years of age or younger, four of 24 (17%) with extensive intraductal component developed an ipsilateral breast relapse, compared with six of 80 (8%) without extensive intraductal component, in contrast to eight of 159 (5%) and 33 of 776 (4%) in postmenopausal patients with or without extensive intraductal component, respectively. In patients with T2 tumors, two of eight (25%) women 40 years or younger with extensive intraductal component, and seven of 50 (14%) without extensive intraductal component developed ipsilateral breast relapse. The corresponding values for the patients older than 40 years were five of 48 (10%) and 13 of 202 (6%), respectively. The incidences of ipsilateral breast relapses, correlated with status of surgical margins after re-excision in T1 tumors, were one of 30 (3.3%) for positive, no relapses in 40 patients with close margins, 16 of 438 (3.6%) for negative, and 18 of 196 (9%) for undetermined margins. In the patients with T2 tumors, ipsilateral breast relapses occurred in two of 16 patients (12.5%) with positive margins, one of 16 (6%) with close, seven of 105 (6.6%)with negative, and four of 68 (5.9%) with undetermined margins (differences not statistically significant). In patients with T1 tumors, negative margins, the 10-year relapse rate was the same (8%) in 559 to whom a boost was administered and in 66 without a boost. In patients with positive margins, the relapse rate was 4% in 215 receiving a boost (18-20 Gy) and 33% (two of six) without a boost. In patients with T2 tumors and negative margins, the rate of ipsilateral breast relapses in 16 patients to whom no boost was given was 12%, as opposed to 10% in 143 patients who received a boost. However, with T2 tumors and close or positive margins, the IBR rate at 10 years was 12% in 81 given a boost, in contrast to 40% (2 of 5) without a boost. In T1 tumors, the breast failure rate was two of 53 (3.7%) in women < or = 40 years receiving chemotherapy and eight of 51 (15.6%) without chemotherapy. For T2 tumors, the corresponding values were seven of 39 (17%) and two of 19 (10.5%), respectively. In women 40 years or younger with T1 tumors receiving hormones or not, the ipsilateral breast relapse rate was two of 19 (10.5%) and eight of 85 (9.4%), respectively; in the older than 40 years group, the corresponding values were six of 377 (1.6%) and 35 of 558 (6.2%). In the patients with T2 tumors, ipsilateral breast relapse rates were not statistically different in the various groups. On multivariate analysis, only age and adjuvant therapy were significant factors predictive of ipsilateral breast relapse. CONCLUSIONS: Surgical excision margins status following adequate doses of radiation therapy was not a predictor of ipsilateral breast relapse. In patients younger than 40 years of age with extensive intraductal component, a somewhat higher breast relapse rate was noted but not enough to preclude breast conservation therapy. A boost of irradiation did not have a significant impact in the incidence of ipsilateral breast relapse in patients with negative margins, but it was of benefit to those with close or positive margins. Close attention to surgical margin status and delivery of higher doses of irradiation to the tumor excision site in patients with close or positive surgical margins will decrease the probability of breast relapses. PMID- 14740973 TI - Predictors of extracapsular extension and its radial distance in prostate cancer: implications for prostate IMRT, brachytherapy, and surgery. AB - PURPOSE: Tightly constricted isodose lines are generated using brachytherapy or intensity-modulated radiation therapy (IMRT) treatment planning systems for prostate cancer. Definition of margins that encompass subclinical disease extension is important to maximize dose escalation while attemptingto adhere to normal tissue dose tolerances. In this study, we attempted to find predictors of extracapsular extension (ECE) and its radial distance. MATERIALS AND METHODS: Pathological assessment of ECE and its radial distance was performed on 712 radical prostatectomy specimens. Preoperative data (initial prostate-specific antigen, clinical stage, ultrasound volume, and biopsy Gleason score) were evaluated for their ability to predict the presence of ECE and its radial distance. RESULTS: Measurable disease was noted outside the prostatic capsule in 185 of 712 (26.0%) specimens. All preoperative parameters except ultrasound volume were able to predict the presence of ECE. However, none of them was predictive of the radial ECE distance. In this group, the median and the range of the maximum depth of invasion (radial extension from the capsule) were 2.00 and 0.5-12.00 mm, respectively. The mean radial distance from the capsule was 2.93 mm, SD +/- 2.286 mm. All subgroups had some patients with radial extension ranging from 0-2 mm, 2-5 mm, to > 5 mm. Only patients with a prostate-specific antigen of 0-4 ng/mL had no extension > 5 mm. CONCLUSIONS: This is the largest series in the literature thus far that quantitatively assesses radial extracapsular extension. Coverage of subclinical disease must be addressed carefully before successful implementation of intensity-modulated radiation therapy, brachytherapy, or prostatectomy in order to avoid geographical miss. PMID- 14740974 TI - Matched-control retrospective study of the acute and late complications in patients with collagen vascular diseases treated with radiation therapy. AB - BACKGROUND: Controversy surrounds the potential complication rate of patients with collagen vascular diseases (CVD) after radiation. We assess the acute and late complications (based on Radiation Therapy Oncology Group criteria) by a matched-control retrospective study. PATIRNTS/METHODS: The charts of 12,000 patients treated with radiation therapy at the University of Louisville from 1982 to 2001 were reviewed for CVD. A total of 38 patients with documented CVD were compared with a matched-control group of 38 patients without CVD. Median follow up for patients with CVD was 35 months. The patients were matched on the basis of site treated, age, dose, date of treatment, sex, treatment goal, follow-up, tumor site and histology, therapeutic technique, and general treatment method. The patients with CVD included 21 patients with systemic lupus erythematosus (55%), two with scleroderma (5%), four with Raynaud's phenomena (11%), three with fibromyalgia (8%), three with polymyalgia rheumatica (8%), three with Sjogren's syndrome (8%), and two with polymyositis-dermatomyositis (5%). Twenty-nine patients received curative doses, and nine patients received palliative doses. RESULTS: No difference was observed in the incidence of acute or late complications between the two groups. For CVD and matched-control patients receiving curative doses, the incidence of acute reaction for grade II was 49% versus 58% and for grade III was 7% versus 7%, respectively. The incidence of late reactions for patients with CVD and the matched control patients for grade I was 3% versus 7%, for grade II was 7% versus 3%, and for grade III was 7% versus 7%, respectively. The patients treated with palliation had a similar incidence of acute reaction in the CVD and the matched-control groups. No patients in the CVD or matched-control group had fatal complications. Only patients with scleroderma had a slight increase in acute and late complications. CONCLUSION: This is the largest matched-control study thus far in the literature. In the comparison between the patients with CVD and the matched-control patients, there was no significant difference in the incidence of acute or late complication. However, there was a higher incidence of radiation complications in patients with scleroderma. Importantly, no fatal complication was noted in any of the patients with CVD. PMID- 14740975 TI - Radiation-induced salivary gland tumors: a report of 18 cases and a review of the literature. AB - PURPOSE: The purpose of this study was to examine the relationship between radiation exposure and subsequent development of salivary gland tumors (SGTs). MATERIALS AND METHODS: Eighteen patients with SGTs as second cancers after head and neck irradiation were identified from chart review of institutional databases of 3025 patients with SGTs evaluated between 1986 and 2001. RESULTS: The median age at the time of initial radiation therapy was 22 years (range, 5-74 years). The median age of the group at the time of their diagnosis of a SGT was 54 years (range, 21-79 years). The median interval between radiation exposure and diagnosis of the SGT was 21 years (range, 4-64 years). The most common initial diagnosis for which patients received radiation therapy was Hodgkin's disease. Histology of the secondary SGT was varied, but most were malignant (N = 15), and mucoepidermoid carcinoma was the most common histology (N = 9). CONCLUSION: There is an association between radiation exposure and the risk of developing an SGT. There was a wide range in the dose of radiation and the age at exposure, suggesting that exposure to head and neck radiation at any age and dose may increase the risk for SGT. This series also suggests an increased risk for developing malignant (versus benign) SGT after radiation exposure. PMID- 14740976 TI - Radiotherapy for moderate-to-severe Graves' ophthalmopathy: improved outcomes with early treatment. AB - PURPOSE: The purpose of this review is to quantify the response to radiotherapy delivered early in the active inflammatory phase of moderate-to-severe Graves' ophthalmopathy. MATERIALS AND METHODS: A retrospective review of radiotherapy delivered early in the active phase of Graves' ophthalmopathy was performed. All 47 cases had moderate-to-severe ophthalmopathy, and 30 cases had optic neuropathy. Variables examined included exophthalmos, color vision, and resistance to retropulsion. Statistical analyses comparing these variables at presentation and at 3 and 6 months after radiotherapy were performed. Subset analyses comparing responses of patients with symptoms lasting longer or less than 6 months were also performed. RESULTS: At 6 months after radiotherapy, there was improvement in exophthalmos in 74.5% of cases and improvement in retropulsion in 83.0%, and all cases of previous color deficiency improved. The mean improvement in exophthalmos was 1.38 mm, color vision was two plates, and retropulsion was 1 grade. On subset analyses, there was a trend toward greater improvement in patients treated earlier in the course of their symptoms. CONCLUSIONS: Radiotherapy may play an important role in the treatment of Graves' ophthalmopathy, especially for patients with optic neuropathy. Early intervention (symptoms < 6 months) with radiotherapy may be the optimal timing for this treatment. PMID- 14740978 TI - Lessons learned in bioterrorism can be applied to medical practice. PMID- 14740977 TI - Promising early local control of malignant pleural mesothelioma following postoperative intensity modulated radiotherapy (IMRT) to the chest. AB - PURPOSE: Malignant pleural mesothelioma often recurs locally in spite of aggressive resection by extrapleural pneumonectomy and conventional radiotherapy. This may be due to failure to recognize the extent of clinical target volume (CTV) or suboptimal dose delivery to a target that abuts the heart, esophagus, liver, lung, kidney, and spinal cord. We report how these geometric/dosimetric constraints were overcome by exploiting intensity-modulated radiotherapy in the first cohort patient. MATERIALS AND METHODS: Twenty-eight patients who had undergone extrapleural pneumonectomy were treated with intensity-modulated radiotherapy. The CTV included the surgically violated inner chest wall, insertion of diaphragm, pleural reflections, and deep margin of the incision. CTV delineation was facilitated by intraoperative radio-opaque marking. Motion was assessed. CTV doses were 45-50 Gy with boosts taken to 60 Gy. RESULTS: Despite the large, irregular CTV (median, 4151 cc; range, 2667-7286 cc), an average of 97% of the CTV was covered to the target dose (range, 92%-100%). Respiratory motion was minimal because of immobility of the prosthetic diaphragm. Normal tissue dose constraints were met. The commonest effects were nausea/vomiting (89%) and dyspnea (80%). Esophagitis was absent (59% of patients) or mild (34% grade 1/2). At median follow-up of 9 months (range, 5-27 months), local control within the contoured target was 100%. One-year survival, disease-specific survival, and disease-free survival are 65%, 91%, and 88%, respectively. CONCLUSIONS: Intensity-modulated radiotherapy after extrapleural pneumonectomy is tolerable and seems effective, at least at this early point. As local control improves, systemic metastases become more common, and it may be appropriate to add novel agents to further improve the therapeutic ratio. PMID- 14740979 TI - Support needed for clinical faculty in osteopathic emergency medicine residencies. PMID- 14740980 TI - Osteopathic manipulative treatment in prenatal care: a retrospective case control design study. AB - The use of osteopathic manipulative treatment (OMT) during pregnancy has a long tradition in osteopathic medicine. A retrospective study was designed to compare a group of women who received prenatal OMT with a matched group that did not receive prenatal OMT. The medical records of 160 women from four cities who received prenatal OMT were reviewed for the occurrence of meconium-stained amniotic fluid, preterm delivery, use of forceps, and cesarean delivery. The randomly selected records of 161 women who were from the same cities, but who did not receive prenatal OMT, were reviewed for the same outcomes. The results of a logistic regression analysis were statistically reliable, chi2 (4, N = 321) = 26.55; P < .001, indicating that the labor and delivery outcomes, as a set, were associated with whether OMT was administered during pregnancy. According to the Wald criterion, prenatal OMT was significantly associated with meconium-stained amniotic fluid (Z = 13.20, P < .001) and preterm delivery (Z = 9.91; P < .01), while the use of forceps was found to be marginally significant (Z = 3.28; P = .07). The case control study found evidence of improved outcomes in labor and delivery for women who received prenatal OMT, compared with women who did not. A prospective study is proposed as the next step in evaluating the effects of prenatal OMT. PMID- 14740981 TI - Review of integrated neuromusculoskeletal release and the novel application of a segmental anterior/posterior approach in the thoracic, lumbar, and sacral regions. AB - Integrated neuromusculoskeletal release (INR) using a segmental anterior/posterior approach is an osteopathic manipulative treatment technique that is easily learned and applied. The segmental anterior/posterior approach to INR was developed as a practical osteopathic manipulative treatment procedure for the inpatient setting, but also has equal efficacy in the outpatient setting. It builds on the principles of INR and myofascial release techniques, as well as other techniques. This approach focuses on both the anterior and posterior connectivity of the body through the neuromusculoskeletal system and uses this connectivity to effectively treat somatic dysfunctions. The principles of INR are discussed, as well as the role of INR in the diagnosis and treatment of somatic dysfunctions in the thoracic, lumbar, and sacral regions. PMID- 14740982 TI - Primary care physicians' attitudes and practices regarding complementary and alternative medicine. AB - Data were gathered from 423 osteopathic primary care physicians who are members of the Michigan Osteopathic Association, assessing their attitudes and practices regarding complementary and alternative medicine (CAM). Family physicians and general internists were more likely than pediatricians to talk to their patients about CAM. Similarly, female physicians were more likely than male physicians to talk to their patients about CAM or refer their patients for CAM. Finally, physicians aged 35 years and younger were more likely than those 60 years and older to use CAM for themselves or their families. Predominant among the conditions for which the physicians would refer for CAM were long-term problems, traditional therapy failures, psychiatric disorders, and behavioral problems. Results reveal wide variations in the way osteopathic primary care physicians view and use complementary and alternative care. PMID- 14740983 TI - Bedside cardiology skills training for the osteopathic internist using simulation technology. AB - Medical practice changes that limit patient availability, instructor time, and advances in technology have led to a greater use of simulators and multimedia computers in medical education. These systems address the problem of inadequate bedside skills training and poor proficiency among all health care providers. While studies have shown their effectiveness among medical students, residents, and practicing physicians, none has focused on the osteopathic internist population--one that is becoming more responsible for conducting initial and follow-up physical examinations. This report describes the use of "Harvey," the cardiology patient simulator, and the UMedic Multimedia Computer System at a workshop conducted at the American College of Osteopathic Internists' 61st Annual Convention and Scientific Sessions. Participants in this study significantly improved their ability to identify common cardiac auscultatory events, as indicated by pretest-to-posttest scores. Workshop participants were nearly unanimous in their belief that they would like to use these tools for learning and assessment. PMID- 14740984 TI - Metal contamination of sediments and soils of Bayou Saint John: a potential health impact to local fishermen? AB - This research examines the pattern of sediment contamination of an urban bayou of New Orleans (formerly a natural waterway) and the potential for human exposure from consumption of fish caught in the bayou. Sediments and soils of Bayou Saint John were evaluated for lead (Pb), zinc (Zn), and cadmium (Cd). Sediment cores were collected at bridges (n = 130) and sites between the bridges (n = 303) of the bayou. In addition, soil samples (n = 66) were collected along the banks of the bayou. Sediments below the bridges contain significantly more (p-value approximately 10(-7)) Pb and Zn (medians of 241 and 230 mg kg(-1), respectively) than bayou sediments located between bridges (medians of 64 and 77 mg kg(-1), respectively). Sediments below bridges of the upper reaches of the bayou contain significantly larger amounts of metals (p < 10(-14) for Pb and Zn and p approximately 10(-8) for Cd) (medians of 329, 383 and 1.5, respectively) than sediments below bridges in the lower reaches of the bayou (medians of 43, 31 and 0.5 for Pb, Zn and Cd, respectively). Likewise, medians for sediments located between bridges contain significantly (p < 10(-14)) higher quantities of Pb, Zn and Cd (170, 203 and 1.8 mg kg(-1), respectively) in the upper bayou than Pb, Zn, and Cd (48, 32, and 0.8 mg kg(-1), respectively) in the lower reaches of the bayou. The potential risk for human exposure may be magnified by the fact that fishing generally occurs from the numerous bridges that cross the bayou. Poor and minority people do most of the fishing. Most people (87%) indicated they ate fish they caught from the Bayou. PMID- 14740985 TI - Urban geochemistry: a multimedia and multielement survey of a small town in northern Europe. AB - The concentrations and distributions of chemical elements (Ag, Al, As, Au, Ba, Bi, Ca, Cd, Co, Cr, Cu, Fe, Ga, Hg, K, La, Mg, Mn, Mo, Na, Ni, P, Pb, S, Sb, Sc, Se, Sr, Te, Th, Ti, Tl, U, V, W and Zn) were studied in till, humus and urban soil in Jakobstad, a small town (20,000 inhabitants) in W. Finland. The analyses were performed with ICP-MS after aqua regia leaching of till (n = 37), urban topsoil (n = 32) and subsoil (n = 32), and HNO3 leaching of humus (n = 37). The till and humus samples, collected at the same sites, were divided into urban and rural samples. The urban till was not significantly enriched in metals. In contrast, a majority of the elements occurred in higher concentrations in the urban than the rural humus samples. Statistical and spatial interpretations of the humus data revealed that traffic (Pb, V and Ni), metal industry (Pb, Zn, Bi, Sb and Cr), an abandoned shooting range (Pb and Sb) and other sources contribute to higher metal levels in the urban humus. The urban soil samples were collected at parks, yards, abandoned industrial sites, roadsides, etc. The topsoil samples were enriched in most elements, also by elements not enriched in the urban humus (e.g. Cd). At several sites, the concentrations far exceeded the limit concentrations for contaminated soils in Finland. A large variety of sources were identified or indicated. PMID- 14740986 TI - Distribution and risk assessment of fluoride in drinking water in the west plain region of Jilin province, China. AB - The west plain region of Jilin province of northeast China is one of the typical endemic fluorosis areas caused by drinking water for many years. Investigations of hydrogeological and ecoenvironmental conditions as well as endemic fluorosis were conducted in 1998. Results show that the ground water, especially, the water in the unconfined aquifer is the main source of drinking water for local residents. The fluoride concentration in groundwater in the unconfined aquifers is higher than that in the confined aquifer in the west plain of Jilin province. The fluoride concentration in the unconfined aquifer can be used to classify the plain into fluoride deficient area, optimum area and excess area, which trend from west to east. High fluoride (>1.0 mg L(-1)) in drinking water resulted in dental and skeletal fluorosis in local residents (children and pregnant women). There exists a positive correlation between fluoride concentration in the drinking water and the morbidities of endemic fluorosis disease (r1 = 0.781, r2 = 0.872). Health risks associated with fluoride concentration in drinking water are assessed. It has been determined that fluoride concentration in excess of 1.0 mg L(-1) exposes residents to high health risks based on risk identification. The study area is classified into five health risk classes as shown in Figure 4. The risk indexes of this area more than 1.0 are accounted for 68% of the total west plain region. PMID- 14740987 TI - Heavy metals in the bed and suspended sediments of Anyang River, Korea: implications for water quality. AB - The objective of this study is to compare Anyang River bed sediments with water chemical composition and to assess the anthropogenic chemical inputs into the river system. Eight sampling locations were chosen along the river channel. Bed and suspended river sediments and water samples were collected, and analyzed for their chemical and physical composition. Data revealed that trace element concentrations in the river water were generally below world average, except for As, Mn, Ni and Cr. Among the three phases: water, bed and suspended sediment, more than 99% of the trace elements was associated with the bed sediment. Concentrations of trace elements in the sediment were a function a particle size distribution and organic content. The calculated degrees of enrichment based on the least influenced sample (ASD 1) indicated the river sediments were enriched with respect to background. The enrichment factors for Pb, Zn and As were relatively lower than for Cr, Co, Ni and Zn. The difference in the enrichment seems to reflect the human activities influence in the basin, and specially for Cd. Speciation of the elements in the five different chemical forms in the sediment by sequential extraction indicated that the reducible fraction was predominant for Fe, Zinc and Cu showed an irregular variation among the different fractions; whereas, Cd and Pb were more regular. Zinc and Cu highly existed mostly in exchangeable forms. Acid soluble and reducible forms were also important for most metals. The speciation implies that the metals associated with the sediment are subject to release into water bodies as goechemical variables (pH and Eh) change. Currently, the introduced metals are deposited near the source area and are mostly associated with the sediment, implying that the river bed sediment acts mainly as a sink, rather than a pool. The accumulated and enriched toxic trace elements can pose a potential pollution of river water. PMID- 14740988 TI - Spatial and seasonal variations of water quality in a Mediterranean catchment: the Llobregat River (NE Spain). AB - The Llobregat and Ter Rivers, typical Mediterranean catchments in Northeast Spain, supply water to more than 4.5 million inhabitants residing in the metropolitan area of Barcelona. The objective of this work is to study the factors that influence the surface water quality of Llobregat catchment. As such, spatial and temporal variations of more 50 water chemical parameters were monitored in 10 sampling sites for a period that extended from July 1996 to December 2000. The temperature, pH and conductivity were measured at sites, whereas metals were analysed using ICP-OES and ICP-MS instrumental techniques. The head waters of the Llobregat River catchment flow through detrital Mesozoic Cenozoic sedimentary rocks resulting in calcium bicarbonate-type water with low mineral content. The high water quality of the waterhead is deteriorated in the upper-middle part of the catchment due to: occurrence of evaporite-bearing geological formations, and the mining and industrial activities related to potash exploitation. As a result, an obvious increase in Na, K, Mg, Cl-, Br, Rb, and Sr concentrations is reported leading to a sodium (potassium) chloride water type. This saline hydrochemical fingerprint persists downstream. This important feature renders the low water quality of the Llobregat River to be adequate for drinking supply purposes. In addition, the industrial and residential activities, specially at the lower part of the catchment, increases P, B, Mn, Fe, Pb, Al, Cr, Co, Ni, Cu, Zn, As and Sb water concentrations. PMID- 14740989 TI - Potential fluoride contamination in the drinking water of Naranji area, NorthWest Frontier Province, Pakistan. AB - The drinking water of Naranji village and surrounding areas of the NorthWest Frontier Province (NWFP), Pakistan, was analyzed for its fluoride content. The fluoride content of water samples from the springs in the Naranji village goes up to 13.52 mg kg(-1), which exceeds the permissible limit (1.5 mg kg(-1)) set by the World Health Organization (WHO). This study explores the source of fluoride contamination and attributes it to the alkaline rocks of the Ambela granitic complex and the Koga complex. The low-lying areas towards the south have a fluoride content within the permissible limit. The Naranji area therefore needs urgent remedial measures. PMID- 14740990 TI - Fluoride content in bones of Adelie penguins and environmental media in Antarctica. AB - Fluoride (F) distribution and its effects (fluorosis) were investigated in Antarctica. Droppings (L) excreta selected of aquatic birds, lake water, soil and moss (Polytrichum alpinum) showed a high F concentration. Although bones of Adelie penguin (Pygiscelis adeliae) and skua (Catharacta maccormicki) showed exceptionally very high F concentration in the range of 832 to 7187 mg kg(-1), their radiographs did not show any evidence of skeletal fluorosis. The possible reason and geochemical aspects of F in Antarctica region are discussed. PMID- 14740991 TI - Introductory ophthalmic genetics. AB - Rapid progress is occurring in molecular cell biology and genetics in the understanding of basic cellular mechanisms and the potential for genetic therapy. As new methods of genetic prognosis and treatment become available, and diseases are redefined in genetic terms, it is essential that clinicians know more about genetic therapy. This article provides a basic outline of gene therapy. PMID- 14740992 TI - The genetic loci of open-angle glaucoma. AB - As Posner stated in 1949, the bottom line "to the patient and to his family is..., whether his disease will follow a mild course or will lead to blindness". The final goal of genetic research is the identification of the causal genes in the patient, to aid the ophthalmologist in predicting the outcome, in determining diligent treatment is required, and ultimately, in providing the tools for preventing blindness. PMID- 14740993 TI - Current perspectives on the TIGR/MYOC gene (Myocilin) and glaucoma. AB - Over the past several years, many groups worldwide have confirmed the presence of probable disease-causing mutations in the coding region of the (TIGR/MYOC) gene associated with glaucoma. Disease-associated point mutations are often found in the third exon of TIGR/MYOC and are predicted to exert a substantial influence on protein structure. Although there has been speculation as to the mechanisms involved in the pathogenic effects for a number of the mutations, the processes leading to the development of glaucoma involving TIGR/MYOC remain to be elucidated. In addition to ongoing mutation studies, efforts are underway to follow up on TIGR/MYOC gene regulation studies in human trabecular meshwork cells and other possibly relevant cell types. Potentially by altering gene regulation, a major variant (-1000 G/C), present in 15-20% of individuals, appears to be associated with a more rapid progression of glaucomatous disease. This article addresses several of these areas of research on the TIGR/MYOC gene and glaucoma, briefly presenting currently available evidence and considering or updating information presented previously. PMID- 14740994 TI - Optineurin in primary open angle glaucoma. AB - The authors' initial estimate indicated that mutations in Optineurin are responsible for a significant proportion of LPG/POAG families. Currently, there are up to 1.2 million persons with LPG and up to 2.47 million persons with POAG in the United States alone. Perhaps twice as many individuals are already affected with this condition without any identifiable clinical signs or symptoms. Investigators are eagerly awaiting confirmation of OPTN mutations in other glaucoma populations. Although additional mutations have already been identified in the sporadic cases of LPG, the significance of this gene in high-pressure POAG requires more intensive investigation. Limited data on partial screening of this gene indicate that OPTN mutations are responsible for a limited number of cases of high-pressure POAG. If the reported mutation rates of OPTN in the LPG group can be confirmed in other LPG or POAG patients, then this gene would be useful in diagnosing presymptomatic persons many decades before they develop this silent and blinding eye condition. Early identification of such at-risk patients would provide an opportunity for immediate targeted medical treatments and specific glaucoma therapy that might significantly delay or completely stop the gradual progression of this condition. Therefore, identification of glaucoma-causing genes such as Myocilin, Optineurin, and others could provide molecular diagnostic tools for this category of optic neuropathy. Although patients with advanced glaucoma will not directly benefit from the use of such molecular diagnostic tools, their immediate family members could certainly benefit from the identification of the cause of the glaucoma decades before the first manifestation of the disease. In summary, a series of mutations in the Optineurin gene have been shown to be the principal cause of adult-onset LPG/POAG phenotype in certain pedigrees. The exact mechanisms through which these mutations lead to the development of glaucoma require additional functional study. The existing evidence suggests that direct interaction of Optineurin with E3-14.7K protein probably utilizes TNF-alpha or Fas-ligand pathways to mediate apoptosis, inflammation, or vasoconstriction. Optineurin also functions through its interactions with other proteins in cellular morphogenesis and membrane trafficking (RAB8), vesicle trafficking (Huntingtin), transcription activation (TFIIIA), and assembly or activity of two unknown kinases. Identification of Optineurin as an adult-onset glaucoma gene and its known interaction with a group of proteins provides the first opportunity to study biochemical pathways that are thought to be involved in causation of this group of eye disorders. Furthermore, identification of this gene as a contributing factor to the development of glaucoma gives a useful tool for screening of this disorder in the elderly population and other high-risk individuals. The exact impact of OPTN in the development of all glaucoma phenotypes requires future study. PMID- 14740995 TI - Genetics and biochemistry of primary congenital glaucoma. AB - Several observations noted by early investigators supported the supposition that in most cases, congenital glaucoma is determined by genetic factors. The genetic heterogeneity of PCG was confirmed by genetic linkage studies conducted in the 1990s when the authors determined that CYP1B1 is the congenital glaucoma gene at the GLC3A locus. The coding sequence of CYP1B1 has been subjected to extensive screening in familial and sporadic cases of glaucoma from numerous countries and from a large number of ethnic groups. These studies have provided evidence for extensive allelic heterogeneity at the GLC3A locus. This article also discusses the molecular evidence for reduced penetrance in congenital glaucoma and the phenotypic heterogeneity of CYP1B1 mutations, mouse models of CYP1B1, and the biochemistry of CYP1B1. PMID- 14740996 TI - Genetics of uveitis. AB - Uveitis phenotypes can differ substantially, and most uveitis diseases are considered polygenic with complex inheritance patterns. When considering the genetics of these diseases, common threads can be identified. For example, in virtually every polygenic disease studied, there exists an HLA genetic association. This association can be strong, such as the associations of HLA-B27 with AAU and HLA-A29 with BSCR; or it can be more subtle, involving several HLA genes or a combination of HLA genes that compose specific haplotypes. In many of these conditions, it is hypothesized that genes other than classic MHC genes but located at the MHC locus may be important susceptibility genes. Genome-wide scans and other genetic methods are becoming increasingly successful in identifying genetic loci and candidate genes in many inflammatory disorders that have an uveitic component. It will be important to test these findings as uveitis specific genetic factors. Therefore, the burgeoning understanding of the human genome promises to result in new insight into the pathogenesis of uveitis. PMID- 14740997 TI - Genetics of age-related macular degeneration. AB - Gene discoveries will lead to more effective them pies for AMD by identifying specific underlying disease mechanisms that might be corrected by drugs or gene therapy. For example, investigations are currently being carried out using pigment epithelium-derived factor (FEDF). The gene for this potent inhibitor of angiogenesis has been incorporated into an adenoviral vector and delivered into the eye by intravitreal injection to inhibit growth of new blood vessels in eyes with neovascular AMD. In the future, as the genetics of this complex disease are unraveled, more effective treatments and preventative measures that target specific molecular defects underlying the development of AMD can be expected. PMID- 14740998 TI - Gene therapy and control of angiogenesis. AB - The technology of gene therapy may be one step ahead of the present understanding of angiogenesis. The results of studies to modulate angiogenesis have been encouraging, however, and several of these studies are in the preclinical and clinical phases of testing. It is likely, therefore, that gene therapy for such diseases as diabetes mellitus and age-related macular degeneration will soon become a practical reality. It must be hoped that the lessons learned will be beneficial to others attempting to modulate angiogenesis in systemic disorders such as cancer and rheumatoid arthritis. PMID- 14740999 TI - Treatment of retinal and choroidal degenerations and dystrophies: current status and prospects for gene-based therapy. AB - Inherited retinal and choroidal degenerations account for a significant portion of blindness in children and young adults. This article reviews the current status and future prospects for the treatment of these disorders. Current treatment strategies include nutritional intervention for gyrate atrophy of the choroid and retina with hyperornithinemia, abetalipoproteinemia, and Refsum's disease, as well as vitamin A supplementation for retinitis pigmentosa. Future therapeutic prospects include gene therapy for both recessive and dominant disease, secondary gene-based therapies, such as pharmaceutic gene product replacement and treatment with survival factors, anti-apoptotic agents, and calcium blockers, and, finally, stem cell therapy. PMID- 14741000 TI - Survey of genetic neuro-ophthalmic disorders. AB - This article provides an overview of the genetic aspects of neuro-ophthalmic disorders. Leber's hereditary optic neuropathy, optic nerve hypoplasia, Duane's retraction syndrome, congenital nystagmus, and other disorders of neuro ophthalmic interest are discussed. PMID- 14741001 TI - The genetics of keratoconus. AB - Isolated keratoconus with no associations is by far the most common keratoconus presentation seen by a practicing clinician. This article explores the genetics of this most common form of keratoconus. PMID- 14741002 TI - Retinoblastoma and tumor-suppressor gene therapy. AB - Retinoblastoma as a genetic disease is a paradigm for tumor-suppressor gene theory. The RB gene is one of the best-studied tumor-suppressor genes with known key functions in controlling cell proliferation and differentiation. Reconstitution of RB function in RB-deficient tumor cells induces irreversible growth arrest (senescence) and inhibits telomerase activity, simultaneously correcting two of the four defined carcinogenic events in human cells. RB gene therapy has the advantage of selectively killing tumor cells without adverse side effects to normal somatic cells, and efficacy of the therapy has now been demonstrated in vitro and in immunocompetent mouse models. Most preclinical studies of RB gene therapy reported to date have used RB fragments with enhanced cell growth-suppressing function. The clinical success of RB gene therapy of retinoblastoma, however, requires more innovation in vector development. PMID- 14741003 TI - Glaucoma in Ghana, West Africa: clinical features and the role of mutations in Myocilin. AB - POAG is a severely debilitating disease in West Africa. Currently, many patients seem to be misclassified as having POAG who in fact have CACG. Improved examination techniques, including routine gonioscopy, would increase diagnostic accuracy and improve treatment strategies. The incidence of Myocilin mutations in this group of individuals is similar to that found in other populations around the world. Myocilin plays a limited role in the pathogenesis of POAG in this population. PMID- 14741004 TI - Predictive DNA testing for glaucoma: reality in 2003. AB - As new genes and common mutations are identified, DNA testing can be offered. Like clinical testing used in glaucoma, such as IOP, tonography, disc measurements, nerve fiber layer analysis, and the various methods of visual field analysis, well-designed studies are needed to be able to interpret clearly the meaning of abnormal results. To use DNA testing to identify individuals at high risk for glaucoma, it is necessary to have solid evidence with sensitivity and specificity parameters, genotype-phenotype correlations, and information on prevalence and penetrance. These data will have to be replicated in several studies using large, population-matched control groups. Mass screening of glaucoma patients for Myocilin mutations may be worthwhile if 3% to 5% of glaucoma patients will be positive. For comparison, screening all cases of colon cancer for gene mutations involved in hereditary nonpolyposis colorectal cancer is considered feasible and desirable with a yield of only 3%. Recent research has shown the value of early treatment of glaucoma. The cost effectiveness of genetics screening will need to be weighed against the cost of conventional screening and the benefits of early treatment considered. Within glaucoma pedigrees with known mutations, DNA mutation-positive individuals will need more frequent clinical screening, whereas DNA mutation-negative individuals will need less frequent follow-up. It is likely that, for every positive-mutation glaucoma case identified, there will be on average two siblings and two children to test. In addition to the laboratory costs, the costs of counseling, and, in particular, the availability of suitably trained individuals who can correctly interpret these test results, must be considered. The risk and benefits of these measures must be calculated and then balanced with the long-term visual outcome of such a strategy. How could genetic testing alter management in glaucoma? If a family member in a Myocilin pedigree with a severe mutation is negative for the mutation, that individual's risk changes from 50% to that of the general population (ie, -2%), and the frequency of clinical screening can be reduced. There are ethical issues involved in testing, particularly in children, but testing would seem justified in congenital, developmental, and juvenile glaucoma. Issues related to insurance may affect the decision making of some patients. A further consideration, which may regrettably become important in the future, is that of intellectual property and patent issues pertaining to glaucoma gene discovery. In addition to clinical evidence of the value of predictive DNA testing, it is incumbent on those working in the field to evaluate the acceptability of testing to patients and their family members. The authors' experience to date is that predictive DNA testing in glaucoma is well supported in suitable families. As with predictive DNA screening in other ophthalmic conditions, issues relating to insurance, ethics, and confidentiality need to be taken into consideration. Although many of the more recently described genetic associations of POAG require more thorough evaluation, Myocilin gene testing can and should be offered for young-onset severe glaucoma cases with a positive family history. PMID- 14741005 TI - The problem of overfitting. PMID- 14741006 TI - Kekule count in capped zigzag boron-nitride nanotubes. AB - Hemi-B16N16 capped zigzag boron-nitride nanotube is introduced, and its Kekule count is studied. With a bond-allocating and coding scheme, recurrence formulas are established as well as for the case of a hemi-B36N36 capped zigzag nanotube. Numerical results reveal that the Kekule counts increase exponentially with respect to the number of layers in the nanotubes concerned. PMID- 14741007 TI - Molecular similarity analysis and virtual screening by mapping of consensus positions in binary-transformed chemical descriptor spaces with variable dimensionality. AB - A novel compound classification algorithm is described that operates in binary molecular descriptor spaces and groups active compounds together in a computationally highly efficient manner. The method involves the transformation of continuous descriptor value ranges into a binary format, subsequent definition of simplified descriptor spaces, identification of consensus positions of specific compound sets in these spaces, and iterative adjustments of the dimensionality of the descriptor spaces in order to discriminate compounds sharing similar activity from others. We term this approach Dynamic Mapping of Consensus positions (DMC) because the definition of reference spaces is tuned toward specific compound classes and their dimensionality is increased as the analysis proceeds. When applied to virtual screening, sets of bait compounds are added to a large screening database to identify hidden active molecules. In these calculations, molecules that map to consensus positions after elimination of most of the database compounds are considered hit candidates. In a benchmark study on five biological activity classes, hits for randomly assembled sets of bait molecules were correctly identified in 95% of virtual screening calculations in a source database containing more than 1.3 million molecules, thus providing a measure of the sensitivity of the DMC technique. PMID- 14741008 TI - Step-by-step calculation of all maximum common substructures through a constraint satisfaction based algorithm. AB - In this paper we propose a new algorithm for subgraph isomorphism based on the representation of molecular structures as colored graphs and the representation of these graphs as vectors in n-dimensional spaces. The presented process that obtains all maximum common substructures is based on the solution of a constraint satisfaction problem defined as the common m-dimensional space (m< or =n) in which the vectors representing the matched graphs can be defined. PMID- 14741009 TI - Encoding the core electrons with graph concepts. AB - The core electron problem of atoms in chemical graph studies has always been considered as a minor problem. Usually, chemical graphs had to encode just a small set of second row atoms, i.e., C, N, O, and F, thus, graph and, in some cases, pseudograph concepts were enough to "graph" encode the molecules at hand. Molecular connectivity theory, together with its side-branch the electrotopological state, introduced two "ad hoc" algorithms for the core electrons of higher-row atoms based, mainly, on quantum concepts alike. Recently, complete graphs, and, especially, odd complete graphs have been introduced to encode the core electrons of higher-row atoms. By the aid of these types of graphs a double-valued algorithm has been proposed for the valence delta, deltav, of any type of atoms of the periodic table with a principal quantum number n > or =2. The new algorithm is centered on an invariant suggested by the hand-shaking theorem, and the values it gives rise to parallel in some way the values derived by the aid of the two old "quantum" algorithms. A thorough comparative analysis of the newly proposed algorithms has been undertaken for atoms of the group 1A-7A of the periodic table. This comparative study includes the electronegativity, the size of the atoms, the first ionization energy, and the electron affinity. The given algorithm has also been tested with sequential complete graphs, while the even complete graphs give rise to conceptual difficulties. QSAR/QSPR studies do not show a clear-cut preference for any of the two values the algorithm gives rise to, even if recent results seem to prefer one of the two values. PMID- 14741010 TI - Partitioning of pi-electrons in rings of polycyclic benzenoid hydrocarbons. 2.1. Catacondensed coronoids. AB - Various types of coronoids are examined for finding the partitioning of pi electrons in individual benzenoid rings of polycyclic aromatic hydrocarbons. A class of catacondensed coronoids, namely fibonacenes, has four pi-electrons in every ring. Most other catacondensed coronoids have more than one type of benzenoid ring, each type with a different partition, but the average per ring is always four pi-electrons. For the class of catacondensed coronoids that have an acenic hole, simple quadratic equations in terms of their number R of rings exist for the partitions of pi-electrons in the three possible ring types. PMID- 14741011 TI - Table of periodic properties of fullerenes based on structural parameters. AB - The periodic table (PT) of the elements suggests that hydrogen could be the origin of everything else. The construction principle is an evolutionary process that is formally similar to those of Darwin and Oparin. The Kekule structure count and permanence of the adjacency matrix of fullerenes are related to structural parameters involving the presence of contiguous pentagons p, q and r. Let p be the number of edges common to two pentagons, q the number of vertices common to three pentagons, and r the number of pairs of nonadjacent pentagon edges shared between two other pentagons. Principal component analysis (PCA) of the structural parameters and cluster analysis (CA) of the fullerenes permit classifying them and agree. A PT of the fullerenes is built based on the structural parameters, PCA and CA. The periodic law does not have the rank of the laws of physics. (1) The properties of the fullerenes are not repeated; only, and perhaps, their chemical character. (2) The order relationships are repeated, although with exceptions. The proposed statement is the following: The relationships that any fullerene p has with its neighbor p + 1 are approximately repeated for each period. PMID- 14741012 TI - Topological study of the periodic system. AB - We carried out a topological study of the Space of Chemical Elements, SCE, based on a clustering analysis of 72 elements, each one defined by a vector of 31 properties. We looked for neighborhoods, boundaries, and other topological properties of the SCE. Among the results one sees the well-known patterns of the Periodic Table and relationships such as the Singularity Principle and the Diagonal Relationship, but there appears also a robustness property of some of the better-known families of elements. Alkaline metals and Noble Gases are sets whose neighborhoods have no other elements besides themselves, whereas the topological boundary of the set of metals is formed by semimetallic elements. PMID- 14741013 TI - Implementing the Fisher's discriminant ratio in a k-means clustering algorithm for feature selection and data set trimming. AB - The Fisher's discriminant ratio has been used as a class separability criterion and implemented in a k-means clustering algorithm for performing simultaneous feature selection and data set trimming on a set of 221 HIV-1 protease inhibitors. The total number of molecular descriptors computed for each inhibitor is 43, and they are scaled to lie between 1 and 0 before being subjected to the feature selection process. Since the purpose is to select some of the most class sensitive descriptors, several feature evaluation indices such as the Shannon entropy, the linear regression of selected descriptors on the pKi of selected inhibitors, and a stepwise variable selection program are used to filter them. While the Shannon entropy provides the information content for each descriptor computed, more class sensitive descriptors are searched by both the linear regression and stepwise variable selection procedures. The inhibitors are divided into several different numbers of classes. They are subsequently divided into five classes due to the fact that the best feature selection result is obtained by the division. Most of the good features selected are the topological descriptors, and they are correlated well with the pKi values. The outliers or the inhibitors with less class-sensitive descriptor values computed for each selected descriptor are identified and gathered by the k-means clustering algorithm. These are the trimmed inhibitors, while the remaining ones are retained or selected. We find that 44% or 98 inhibitors can be retained when the number of good descriptors selected for clustering is three. The descriptor values of these selected inhibitors are far more class sensitive than the original ones as evidenced by substantial increasing in statistical significance when they are subjected to both the SYBYL CoMFA PLS and Cerius2 PLS regression analyses. PMID- 14741014 TI - Identification of a minimal subset of receptor conformations for improved multiple conformation docking and two-step scoring. AB - Docking and scoring are critical issues in virtual drug screening methods. Fast and reliable methods are required for the prediction of binding affinity especially when applied to a large library of compounds. The implementation of receptor flexibility and refinement of scoring functions for this purpose are extremely challenging in terms of computational speed. Here we propose a knowledge-based multiple-conformation docking method that efficiently accommodates receptor flexibility thus permitting reliable virtual screening of large compound libraries. Starting with a small number of active compounds, a preliminary docking operation is conducted on a large ensemble of receptor conformations to select the minimal subset of receptor conformations that provides a strong correlation between the experimental binding affinity (e.g., Ki, IC50) and the docking score. Only this subset is used for subsequent multiple conformation docking of the entire data set of library (test) compounds. In conjunction with the multiple-conformation docking procedure, a two-step scoring scheme is employed by which the optimal scoring geometries obtained from the multiple-conformation docking are re-scored by a molecular mechanics energy function including desolvation terms. To demonstrate the feasibility of this approach, we applied this integrated approach to the estrogen receptor alpha (ERalpha) system for which published binding affinity data were available for a series of structurally diverse chemicals. The statistical correlation between docking scores and experimental values was significantly improved from those of single-conformation dockings. This approach led to substantial enrichment of the virtual screening conducted on mixtures of active and inactive ERalpha compounds. PMID- 14741015 TI - CoMFA modeling of human catechol O-methyltransferase enzyme kinetics. AB - Three-dimensional QSAR models with different charge calculation methods (MOPAC AM1-ESP, MOPAC-AM1-Coulson and Gasteiger-Huckel) were developed for predicting all three enzyme kinetic parameters Km, Vmax and Vmax/Km for catecholic substrates of human soluble catechol O-methyltransferase (S-COMT). The empirical parameters of 45 substrates were correlated to the steric and electronic molecular fields of the substrates utilizing Comparative Molecular Field Analysis (CoMFA). Alignment rules for CoMFA were developed based on the catalytic mechanism and crystal structure of S-COMT, and the analysis was optimized using an all-space search technique. Leave-one-out and leave-n-out cross-validation (with 5 and 10 cross-validation groups) was carried out, and all developed models proved to be statistically significant with q2 values up to 0.84. The models based on MOPAC charge calculations predicted the empirical values clearly better than the Gasteiger-Huckel method. The derived CoMFA coefficient contour maps of steric and electrostatic interactions correlated clearly with the S-COMT crystallographic structures. PMID- 14741016 TI - QSAR in ecotoxicity: an overview of modern classification techniques. AB - This study deals with classification for toxicity prediction. Using a data set of 235 pesticides and 153 descriptors, we built several models using seven classification algorithms: nearest mean classifier, linear discriminant analysis, quadratic discriminant analysis, regularized discriminant analysis, soft independent modeling of class analogy, K nearest neighbors classification, classification, and regression tree. The performance of the models was then compared with the classifier, the end-points, the number of descriptor, and the diversity of the data set. Finally, we made a critical analysis of the models and descriptors. PMID- 14741017 TI - Use of variable selection in modeling the secondary structural content of proteins from their composition of amino acid residues. AB - The possibility of prediction of protein secondary structure content from composition of their amino acid residues can help in bridging the gap between proteins of known primary sequence having an unknown secondary structure. Almost all recently published models for understanding the relationship between composition (frequency of occurrence) of amino acid residues and secondary structure content of proteins involved composition of all 20 amino acid residues. However, it is well-known that many amino acid residues are mutually similar according to their physicochemical properties (hydrophobicity, hydrophilicity, charge, size, etc.). Because of that, we were motivated to investigate the possibility of reduction of the total number of terms (frequencies of amino acid residues) in the models for describing the relation between the composition of amino acid residues and the percentage of residues belonging to alpha, beta, and coil secondary structure. For this purpose, the CROMRsel algorithm (J. Chem. Inf. Comput. Sci. 1999, 39, 121-132) for selection of a small subset of the most important variables/descriptors into the multiregression (MR) models, i.e., frequency of occurrence of amino acid residues in proteins, was used. Analysis was performed on a data set containing 475 proteins, taken from Proteins 1996, 25, 157-168. A complete data set was partitioned into a 317-protein training set and 158-protein test set. The best possible linear models containing I=1, ..., 20 frequencies were selected among all 20 frequencies of occurrence of amino acid residues on the 317-protein training set, and were used for performing prediction of the corresponding percentage of secondary structure content on the 158-protein test set. For the 317-protein data set the best selected concise models for the alpha, beta, and coil secondary structure contain only 9, 5, and 8 frequencies, respectively. Selected concise models are of the same or better fitted, cross validated, and predictive statistical parameters than the models containing all 20 frequencies. Additionally, for each I (I=1, ...., 20) 30 the best possible random models were selected. In each case, the best possible real models are much better than each of the best possible random models, showing clearly that there is no risk of a chance correlation (what one could expect due to the application of an exhaustive search for the best model having I frequencies among all 20!/I!(20-I)! possible models). Finally, the best selected models on the complete 475-protein data set for the alpha, beta, and coil secondary structure contain only 7, 4, and 7 frequencies of amino acid residues, respectively. These models are much simpler and have better fitted and cross-validated errors than the corresponding models from the literature, that were obtained without using a procedure for selection of the most important frequencies of amino acid residues in proteins. PMID- 14741018 TI - Topology, connectivity, and electronic structure of C and B cages and the corresponding nanotubes. AB - After a brief discussion of the structural trends which appear with an increasing number of atoms in B cages, a one-to one correspondence between the connectivity of B cages and C cage structures will be proposed. The electronic level spectra of both systems from Hartree-Fock calculations is given and discussed. The relation of curvature introduced into an originally planar graphitic fragment to pentagonal "defects" such as are present in buckminsterfullerene is also briefly treated. A study of the structure and electronic properties of B nanotubes will then be introduced. We start by presenting a solution of the free-electron network approach for a "model boron" planar lattice with local coordination number 6. In particular the dispersion relation E(k) for the pi-electron bands, together with the corresponding electronic Density Of States (DOS), will be exhibited. This is then used within the zone-folding scheme to obtain information about the electronic DOS of different nanotubes obtained by folding this model boron sheet. To obtain the self-consistent potential in which the valence electrons move in a nanotube, "the March model" in its original form was invoked, and the results are reported for a carbon nanotube. Finally, heterostructures, such as BN cages and fluorinated buckminsterfullerene, will be briefly treated, the new feature here being electronegativity difference. PMID- 14741019 TI - Aqueous biphasic systems. Partitioning of organic molecules: a QSPR treatment. AB - The partitioning of 29 small organic probes in a PEG-2000/(NH4)2SO4 biphasic system was investigated using a quantitative structure-property relationship (QSPR) approach. A three-descriptor equation with the squared correlation coefficient (R2) of 0.97 for the partition coefficient (log D) was obtained. All descriptors were derived solely from the chemical structure of the compounds. Using the same descriptors, a three-parameter model was also obtained for log P (octanol/water, R2=0.89); predicted log P values were used as an external descriptor for modeling log D. PMID- 14741020 TI - Efficient discovery of nonlinear dependencies in a combinatorial catalyst data set. AB - Exploration of a complex catalyst system using Genetic Algorithm methods and combinatorial experimentation efficiently removes noncontributing elements and generates data that can be used to model the remaining system. In particular the combined methods effectively navigate and optimize systems with highly nonlinear dependencies (3-way and higher interactions). PMID- 14741021 TI - Development of neural network QSPR models for Hansch substituent constants. 1. Method and validations. AB - In an attempt to develop predictive models for Hansch substituent constants for less common substituents, neural network QSPR (Quantitative Structure-Property Relationship) studies were conducted to correlate Hansch substituent constants for hundreds of chemically diverse functional groups with two different molecular descriptor sets. The Hansch substituent constants under study were pi, MR, F and R, describing the hydrophobic, steric/polarizability, and electronic (field and resonance) characteristics of the substituents, respectively. E-state descriptors were used for pi and MR, while the molecular descriptor set based upon the approach of Kvasnicka, Sklenak, and Pospichal (J. Am. Chem. Soc. 1993, 115, 1495 1500) was adopted for F and R. Both QSPR models demonstrated good predictivity in test sets. PMID- 14741022 TI - Development of neural network QSPR models for Hansch substituent constants. 2. Applications in QSAR studies of HIV-1 reverse transcriptase and dihydrofolate reductase inhibitors. AB - In this paper, the applications of a Hansch substituent constant predictor(1) to Quantitative Structure-Activity Relationships (QSAR) studies of E. coli dihydrofolate reductase (DHFR) inhibitors 2,4-diamino-5-(substituted benzyl)pyrimidines as well as HIV-1 reverse transcriptase (RT) inhibitors 1-[(2 hydroxyethoxy)methyl]-6-(phenylthio)thymine (HEPT) derivatives are demonstrated. Both data sets contain functional groups for which the substituent constants (pi, MR, F and R) could not be found in standard substituent constant tables. The substituent constant predictor allowed us to derive predicted pi, MR, F and R values for all substituents in both data sets, thus enabling the generation of easily interpretable QSAR models of comparable or better predictivity than previous models. PMID- 14741023 TI - Prediction of the isoelectric point of an amino acid based on GA-PLS and SVMs. AB - The support vector machine (SVM), as a novel type of a learning machine, for the first time, was used to develop a QSPR model that relates the structures of 35 amino acids to their isoelectric point. Molecular descriptors calculated from the structure alone were used to represent molecular structures. The seven descriptors selected using GA-PLS, which is a sophisticated hybrid approach that combines GA as a powerful optimization method with PLS as a robust statistical method for variable selection, were used as inputs of RBFNNs and SVM to predict the isoelectric point of an amino acid. The optimal QSPR model developed was based on support vector machines, which showed the following results: the root mean-square error of 0.2383 and the prediction correlation coefficient R=0.9702 were obtained for the whole data set. Satisfactory results indicated that the GA PLS approach is a very effective method for variable selection, and the support vector machine is a very promising tool for the nonlinear approximation. PMID- 14741024 TI - An open source protein gel documentation system for proteome analyses. AB - Data organization and data mining represents one of the main challenges for modern high throughput technologies in pharmaceutical chemistry and medical chemistry. The presented open source documentation and analysis system provides an integrated solution (tutorial, setup protocol, sources, executables) aimed at substituting the traditionally used lab-book. The data management solution provided incorporates detailed information about the processing of the gels and the experimental conditions used and includes basic data analysis facilities which can be easily extended. The sample database and User-Interface are available free of charge under the GNU license from http://webber.physik.uni freiburg.de/~fallerd/tutorial.htm. PMID- 14741025 TI - Molecular similarity searching using atom environments, information-based feature selection, and a naive Bayesian classifier. AB - A novel technique for similarity searching is introduced. Molecules are represented by atom environments, which are fed into an information-gain-based feature selection. A naive Bayesian classifier is then employed for compound classification. The new method is tested by its ability to retrieve five sets of active molecules seeded in the MDL Drug Data Report (MDDR). In comparison experiments, the algorithm outperforms all current retrieval methods assessed here using two- and three-dimensional descriptors and offers insight into the significance of structural components for binding. PMID- 14741026 TI - QSPR modeling of lipid-water partition coefficient by optimization of correlation weights of local graph invariants. AB - The optimization of correlation weights scheme was applied to model lipid-water partition coefficient (log P) of two sets of diverse functional aliphatic and aromatic compounds. In both cases, the optimized descriptors formulated based on the data of training sets generated statistically acceptable relations for the corresponding training sets, test sets, and combined sets. When the relations of log P values with the optimized molecular descriptors formulated based on the data of the training sets were used for calculation of log P values of the corresponding training sets, rpred2 values were found to be satisfactory (above 0.99) in both cases, which is indicative of the predictive potential of the scheme. The results indicate promising potential of the optimization of a correlation weights scheme in modeling studies. PMID- 14741027 TI - Boiling points of halogenated aliphatic compounds: a quantitative structure property relationship for prediction and validation. AB - Halogenated aliphatic compounds have many technical uses, but substances within this group are also ubiquitous environmental pollutants that can affect the ozone layer and contribute to global warming. The establishment of quantitative structure-property relationships is of interest not only to fill in gaps in the available database but also to validate experimental data already acquired. The three-dimensional structures of 240 compounds were modeled with molecular mechanics prior to the generation of empirical descriptors. Two bilinear projection methods, principal component analysis (PCA) and partial-least-squares regression (PLSR), were used to identify outliers. PLSR was subsequently used to build a multivariate calibration model by extracting the latent variables that describe most of the covariation between the molecular structure and the boiling point. Boiling points were also estimated with an extension of the group contribution method of Stein and Brown. PMID- 14741028 TI - Parallel random tunneling algorithm for structural optimization of Lennard-Jones clusters up to N=330. AB - A random tunneling algorithm (RTA) is derived from the terminal repeller unconstrained subenergy tunneling (TRUST) algorithm, and the parallelization of the RTA is implemented with an island parallel paradigm. Combined with the techniques of angular moving, the parallel random tunneling algorithm (PRTA) is applied to the optimization of Lennard-Jones (LJ) atomic clusters, and all the global minima of LJ clusters with size up to 200 are successfully located. For the optimization of larger cluster, a PRTA with an improved seeding technique is developed and successfully applied to the optimization of LJ151-LJ309. Furthermore, the optimized structures of LJ309-330 with the PRTA, which have not been studied before, are also provided. PMID- 14741029 TI - Topological distance based 3D descriptors for use in QSAR and diversity analysis. AB - In topological autocorrelation approaches molecular descriptors are calculated by summing up properties located at given topological distances. Since the relationship between topological and Euclidean distance contains 3D structural information, in the present paper a modified version of an autocorrelation approach is proposed to include this type of information. Steric, electronic, and indicator-variable-type descriptors are calculated and used in QSAR studies with three different data sets. The results demonstrate that the descriptors can be efficiently used in cluster- and QSAR analysis. The models obtained are highly predictive and comparable to those obtained by other commonly used 3D-QSAR methods. PMID- 14741030 TI - A 3D-QSAR of angiotensin II (AT1) receptor antagonists based on receptor surface analysis. AB - A hypothetical receptor surface model has been constructed for a set of 38 AT1 antagonists using activity data of each molecule as a weight in the building of the receptor surface. The best model was derived by optimizing various parameters such as atomic partial charges, surface fit, and the manner of representation of electrostatics on the surface. Descriptors such as van der Waals energy, electrostatic energy, and total nonbonded energy were used individually or in combination to derive a family of quantitative structure-activity relationship equations using G/PLS as the statistical method. PMID- 14741031 TI - Development of a quantitative structure-activity relationship model for inhibition of gram-positive bacterial cell growth by biarylamides. AB - A set of compounds consisting of a new and diverse collection of biarylamides was examined using quantitative structure-activity relationship techniques for the purpose of developing a model to describe inhibition of gram-positive bacterial growth (minimum inhibition concentration). The model was sought in order to obtain insight for designing new molecules. A detailed analysis of the underlying structure-activity relationship helped provide insight concerning which structural features of the molecules modulated the activity of the compounds against gram-positive organisms. PMID- 14741032 TI - Eigenvalue analysis of peroxisome proliferator-activated receptor gamma agonists. AB - Eigenvalue analysis (EVA) was conducted on a series of potent agonists of peroxisome proliferator-activated receptor gamma (PPARgamma). Predictive EVA quantitative structure-activity relationship (QSAR) models were established using the SYBYL package, which had conventional r2 and cross-validated coefficient (q2) values up to 0.920 and 0.587 for the AM1 method and 0.863 and 0.586 for the PM3 method, respectively. These models were validated by a test set containing 18 compounds. The capability to predict by these two models for PPARgamma agonists, with the best predictive r2pred value of 0.614 for AM1 and 0.822 for PM3 methods, set a successful example for applying a similar approach in building QSAR models for PPARalpha and -delta that could potentially offer a new opportunity in the design of novel PPAR modulators. PMID- 14741033 TI - Blood-brain barrier permeation models: discriminating between potential CNS and non-CNS drugs including P-glycoprotein substrates. AB - The aim of this article is to present the design of a large heterogeneous CNS library (approximately 1700 compounds) from WDI and mapping CNS drugs using QSAR models of blood-brain barrier (BBB) permeation and P-gp substrates. The CNS library finally includes 1336 BBB-crossing drugs (BBB+), 259 molecules non-BBB crossing (BBB-), and 91 P-gp substrates (either BBB+ or BBB-). Discriminant analysis and PLS-DA have been used to model the passive diffusion component of BBB permeation and potential physicochemical requirement of P-gp substrates. Three categories of explanatory variables (Cdiff, BBBpred, PGPpred) have been suggested to express the level of permeation within a continuous scale, starting from two classes data (BBB+/BBB-), allowing that the degree to which each compound belongs to an activity class is given using a membership score. Finally, statistical data analyses have shown that some very simple descriptors are sufficient to evaluate BBB permeation in most cases, with a high rate of well classified drugs. Moreover, a "CNS drugs" map, including P-gp substrates and accurately reflecting the in vivo behavior of drugs, is proposed as a tool for CNS drug virtual screening. PMID- 14741034 TI - Web-based tools for mining the NCI databases for anticancer drug discovery. AB - In this paper, we describe the development of a set of integrated Web-based tools for mining the National Cancer Institute's (NCI) anticancer databases for anticancer drug discovery. For data mining, three different correlation algorithms were implemented, which included the commonly used Pearson's correlation algorithm available from the NCI's COMPARE program, the Spearman's and Kendall's correlation algorithms. In addition, we implemented the p-value test to evaluate the significance of the correlation results. These Web-based data mining tools allow robust analysis of the correlation between the in vitro anticancer activity of the drugs in the NCI anticancer database, the protein levels and mRNA levels of molecular targets (genes) in the NCI 60 human cancer cell lines for identification of potential lead compounds for a specific molecular target and for study of the molecular mechanism action of a drug. Examples were provided to identify PKC ligands using a lead compound and to identify potential ErbB-2 inhibitors using the mRNA levels of ErbB-2 in the NCI 60 tumor cell lines. PMID- 14741035 TI - QSAR analysis of the toxicity of aromatic compounds to Chlorella vulgaris in a novel short-term assay. AB - The use of alternative toxicity tests and computational prediction models is widely accepted to fill experimental data gaps and to prioritize chemicals for more expensive and time-consuming assessment. A novel short-term toxicity test using the alga Chlorella vulgaris was utilized in this study to produce acute aquatic toxicity data for 65 aromatic compounds. The compounds tested included phenols, anilines, nitrobenzenes, benzaldehydes and other poly-substituted benzenes. The toxicity data were employed in the development of quantitative structure-activity relationships (QSARs). Using multiple regression (MLR) and partial least squares (PLS) analyses, statistically significant, transparent and interpretable QSARs were developed using a small number of physicochemical descriptors. A two-descriptor model was developed using MLR (log(1/EC50)=0.73 log Kow-0.59 Elumo-1.91; n=65, r2=0.84, r2CV=0.82, s=0.43) and a four-descriptor model using PLS (log(1/EC50)=0.40 log Kow-0.23 Elumo+9.84 Amax+0.20 0chiv-5.40; n=65, r2=0.86, q2=0.84, RMSEE=0.40). The latter model was obtained by stepwise elimination of variables from a set of 102 calculated descriptors. Both models were validated successfully by simulating external prediction through the use of complementary subsets. The two factors, which were identified as being critical for the acute algal toxicity of this set of compounds were hydrophobicity and electrophilicity. PMID- 14741036 TI - ADME evaluation in drug discovery. 4. Prediction of aqueous solubility based on atom contribution approach. AB - A novel method for the estimation of aqueous solubility was solely based on simple atom contribution. Each atom in a molecule has its own contribution to aqueous solubility and was developed. Altogether 76 atom types were used to classify atoms with different chemical environments. Moreover, two correction factors, including hydrophobic carbon and square of molecular weight, were used to account for the inter-/intramolecular hydrophobic interactions and bulkiness effect. The contribution coefficients of different atom types and correction factors were generated based on a multiple linear regression using a learning set consisting of 1290 organic compounds. The obtained linear regression model possesses good statistical significance with an overall correlation coefficient (r) of 0.96, a standard deviation (s) of 0.61, and an unsigned mean error (UME) of 0.48. The actual prediction potential of the model was validated through an external test set with 21 pharmaceutically and environmentally interesting compounds. For the test set, a predictive r=0.94, s=0.84, and UME=0.52 were achieved. Comparisons among eight procedures of solubility calculation for those 21 molecules demonstrate that our model bears very good accuracy and is comparable to or even better than most reported techniques based on molecular descriptors. Moreover, we compared the performance of our model to a test set of 120 molecules with a popular group contribution method developed by Klopman et al. For this test set, our model gives a very effective prediction (r=0.96, s=0.79, UME=0.57), which is obviously superior to the predicted results (r=0.96, s=0.84, UME=0.70) given by the Klopman's group contribution approach. Because of the adoption of atoms as the basic units, our addition model does not contain a "missing fragment" problem and thus may be more simple and universal than the group contribution models and can give predictions for any organic molecules. A program, drug-LOGS, had been developed to identify the occurrence of atom types and estimate the aqueous solubility of a molecule. PMID- 14741037 TI - 2D QSAR consensus prediction for high-throughput virtual screening. An application to COX-2 inhibition modeling and screening of the NCI database. AB - Using classification (SOM, LVQ, Binary, Decision Tree) and regression algorithms (PLS, BRANN, k-NN, Linear), this paper details the building of eight 2D-QSAR models from a 266 COX-2 inhibitor training set. The predictive performances of these eight models were subsequently compared using an 88 COX-2 inhibitor test set. Each ligand is described by 52 2D descriptors expressed as van der Waals Surface Areas (P_VSA) and its COX-2 binding IC50. One of our best predictive models is the neural network model (BRANN), which is able to select a subset, from the 88 ligand test set, that contains 94% COX-2 active inhibitors (pIC50>7.5) and detects 71% of all the actives. We then introduce a QSAR consensus prediction protocol that is shown to be more predictive than any single QSAR model: our C3 consensus approach is able to select a subset from the 88 ligand test set that contains 94% active inhibitors and 83% of all the actives. The 2D QSAR consensus protocol was finally applied to the high-throughput virtual screening of the NCI database, containing 193,477 organic compounds. PMID- 14741038 TI - Chirality of the disulfide in the prion proteins. AB - In the Transmissible Spongiform Encephalopathies (TSEs) it has been generally assumed that the normal prion proteins (PPr) occurring on neural cells have the same composition of amino acids and the same sequence as the pathological forms (PPrSc) but differ in the manner of folding. The mechanism(s) by which the conversion of PPr into PPrSc takes place remain unknown. This paper calls attention to some aspects of chirality inherent in the disulfide function and suggests the possibility that handedness in the disulfide bond of prions may transmit stereochemical information that can influence the manner of folding or refolding into pathogenic forms. PMID- 14741039 TI - Glucose transporter content and glucose uptake in skeletal muscle constructs engineered in vitro. AB - Engineered muscle may eventually be used as a treatment option for patients suffering from loss of muscle function. The metabolic and contractile function of engineered muscle has not been well described; therefore, the purpose of this experiment was to study glucose transporter content and glucose uptake in engineered skeletal muscle constructs called myooids. Glucose uptake by way of 2 deoxyglucose and GLUT-1 and GLUT-4 transporter protein content was measured in basal and insulin-stimulated myooids that were engineered from soleus muscles of female Sprague-Dawley rats. There was a significant increase in the basal 2 deoxyglucose uptake of myooids compared with adult control (fivefold), contraction-stimulated (3.4-fold), and insulin-stimulated (threefold) soleus muscles (P = 0.0001, 0.0001, and 0.0001, respectively). In addition, there was a significant increase in the insulin-stimulated 2-deoxyglucose uptake of myooids compared with adult control soleus muscles in basal conditions (6.5-fold) and adult contraction-stimulated (4.5-fold) and insulin- stimulated (3.9-fold) soleus muscles (P = 0.0001, 0.0001, and 0.0001, respectively). There was a significant 30% increase in insulin-stimulated compared with basal 2-deoxyglucose uptake in the myooids. The myooid GLUT-1 protein content was 820% of the adult control soleus muscle, whereas the GLUT-4 protein content was 130% of the control soleus muscle. Myooid GLUT-1 protein content was 6.3-fold greater than GLUT-4 protein content, suggesting that the glucose transport of the engineered myooids is similar in several respects to that observed in both fetal and denervated skeletal muscle tissue. PMID- 14741040 TI - A novel cell-based system for evaluating skeletal muscle cell hypertrophy inducing agents. AB - Skeletal muscle is a tissue that adapts to increased use by increasing contractile protein gene expression and ultimately skeletal muscle mass (hypertrophy). To identify hypertrophy-inducing agents that may be potentially useful in the treatment of age-related muscle loss (sarcopenia) and to better understand hypertrophy signal transduction pathways, we have created a skeletal muscle cell-based hypertrophy-responsive system. This system was created by permanently modifying the relatively undifferentiated C2C12 cell line so that it contains the beta-myosin heavy chain (beta-MHC) gene promoter and enhancer regions fused to a luciferase reporter gene. This cell line responds, by increasing luciferase expression, to a variety of skeletal muscle hypertrophy inducing agents, including insulin, insulin-like growth factor I, testosterone, and the beta-adrenergic receptor agonist isoproterenol, in both the undifferentiated and differentiated states. This cell-based system should be useful for identifying novel hypertrophy-inducing agents as well as understanding hypertrophy signal transduction. PMID- 14741041 TI - Diet, the endothelium and atherosclerosis. AB - Dietary factors have been implicated in the difference in the risk of cardiovascular disease observed between populations. However, clinical trials and mechanistic studies of dietary modification are difficult to undertake, and the data set on dietary intervention for cardiovascular disease is now much smaller than that for pharmacological interventions. Moreover, the precise mechanism by which dietary modification might alter the risk of cardiovascular disease is uncertain. In this issue of Clinical Science, Ambring and co-workers investigate the effect of a Mediterranean-inspired diet on endothelial function and lipid fractions. PMID- 14741042 TI - Inflammation and endothelial dysfunction: intimate companions in the pathogenesis of vascular disease? AB - There is increasing evidence to implicate inflammation as an important precursor of endothelial dysfunction. This mechanistic link is apparent across the entire spectrum of inflammatory status, i.e. endothelial function is apparent following acute infection, and in subjects with chronic high-grade inflammation and, perhaps most importantly, persistent low-grade inflammation. The recognition of this relationship has present therapeutic ramifications, but also requires that future longitudinal studies determining the predictive ability of endothelial function measures for vascular events should incorporate markers of inflammation as potential confounders. In this issue of Clinical Science, Fichtlscherer and co workers describe a link between endothelial function and sPLA(2) (secretory non pancreatic type II phospholipase A(2)) serum activity. PMID- 14741043 TI - Acute Escherichia coli endotoxaemia decreases the plasma l-arginine/asymmetrical dimethylarginine ratio in humans. AB - Acute inflammation impairs vascular function. Based on the association between endothelial dysfunction and plasma concentrations of L-arginine and the endogenous nitric oxide synthase inhibitor ADMA (asymmetrical dimethylarginine), we hypothesized that the ratio between L-arginine and ADMA could be affected by experimental inflammation. Plasma concentrations of L-arginine, ADMA and SDMA (symmetrical dimethylarginine) were studied at baseline and 3.5 h after intravenous administration of Escherichia coli endotoxin [LPS (lipopolysaccharide), 20 units/kg of body mass; n =8] or placebo ( n =9) in healthy males. L-Arginine and dimethylarginines were quantified after solid-phase extraction by reversed-phase HPLC. Body temperature, heart rate and leucocyte count increased after LPS administration ( P <0.01 for all). LPS administration decreased plasma concentrations of L-arginine from 66 micromol/l [95% CI (confidence interval): 56, 88] at baseline to 48 micromol/l (CI: 40, 60) after 3.5 h ( P <0.02), but did not affect ADMA and SDMA concentrations. Consequently, the L-arginine/ADMA ratio declined significantly from a median of 159 (CI: 137, 193) to 135 (CI: 103, 146); a decrease of 25 (CI: -68, -13; P <0.02). L-Arginine, ADMA, SDMA and the L-arginine/ADMA ratio remained constant over time in controls. Acute inflammation reduces the L-arginine/ADMA ratio which could contribute to impaired vascular function. PMID- 14741044 TI - Identification of furin pro-region determinants involved in folding and activation. AB - The pro-region of the subtilisin-like convertase furin acts early in the biosynthetic pathway as an intramolecular chaperone to enable proper folding of the zymogen, and later on as an inhibitor to constrain the activity of the enzyme until it reaches the trans -Golgi network. To identify residues that are important for pro-region function, we initially identified amino acids that are conserved among the pro-regions of various mammalian convertases. Site-directed mutagenesis of 17 selected amino acids within the 89-residue pro-region and biosynthetic labelling revealed that I60A-furin and H66A-furin were rapidly degraded in a proteasome-dependent manner, while W34A-furin and F67A-furin did not show any autocatalytic activation. Intriguingly, the latter mutants proteolytically cleaved pro-von Willebrand factor precursor to the mature polypeptide, suggesting that the mutations permitted proper folding, but did not allow the pro-region to exercise its role in inhibiting the enzyme. Homology modelling of furin's pro-region revealed that residues Ile-60 and His-66 might be crucial in forming the binding interface with the catalytic domain, while residues Trp-34 and Phe-67 might be involved in maintaining a hydrophobic core within the pro-region itself. These results provide structural insights into the dual role of furin's pro-region. PMID- 14741045 TI - Nuclear factor-kappaB motif and interferon-alpha-stimulated response element co operate in the activation of guanylate-binding protein-1 expression by inflammatory cytokines in endothelial cells. AB - The large GTPase GBP-1 (guanylate-binding protein-1) is a major IFN-gamma (interferon-gamma)-induced protein with potent anti-angiogenic activity in endothelial cells. An ISRE (IFN-alpha-stimulated response element) is necessary and sufficient for the induction of GBP-1 expression by IFN-gamma. Recently, we have shown that in vivo GBP-1 expression is strongly endothelial-cell-associated and is, in addition to IFN-gamma, also activated by interleukin-1beta and tumour necrosis factor-alpha, both in vitro and in vivo [Lubeseder-Martellato, Guenzi, Jorg, Topolt, Naschberger, Kremmer, Zietz, Tschachler, Hutzler, Schwemmle et al. (2002) Am. J. Pathol. 161, 1749-1759; Guenzi, Topolt, Cornali, Lubeseder Martellato, Jorg, Matzen, Zietz, Kremmer, Nappi, Schwemmle et al. (2001) EMBO J. 20, 5568-5577]. In the present study, we identified a NF-kappaB (nuclear factor kappaB)-binding motif that, together with ISRE, is required for the induction of GBP-1 expression by interleukin-1beta and tumour necrosis factor-alpha. Deactivation of the NF-kappaB motif reduced the additive effects of combinations of these cytokines with IFN-gamma by more than 50%. Importantly, NF-kappaB p50 rather than p65 activated the GBP-1 promoter. The NF-kappaB motif and ISRE were detected in an almost identical spatial organization, as in the GBP-1 promoter, in the promoter regions of various inflammation-associated genes. Therefore both motifs may constitute a cooperative inflammatory cytokine response module that regulates GBP-1 expression. Our findings may open new perspectives for the use of NF-kappaB inhibitors to support angiogenesis in inflammatory diseases including ischaemia. PMID- 14741047 TI - Different subcellular localization of sulphotransferase 2B1b in human placenta and prostate. AB - The human hydroxysteroid SULT (sulphotransferase) 2B1 subfamily consists of two isoforms, SULT2B1a and SULT2B1b. These two isoenzymes are transcribed from the same gene by alternative splicing of their first exons and share 94% amino acid sequence identity. The SULT2B1 isoforms are highly selective for the sulphation of 3beta-hydroxysteroids. Immunoblot analysis of SULT2B1 expression in several human tissues indicates the presence of only SULT2B1b protein. Immunoreactive SULT2B1b protein was detected in human prostate, skin, placenta and lung tissue. SULT2B1b mRNA expression was detected in RNA isolated from term placenta, normal prostate, prostate carcinoma, benign prostate hyperplasia, LNCaP prostate cancer cells, breast cancer specimens and MCF-7 breast cancer cells. Immunohistochemical localization of SULT2B1b, in terms placental and prostate tissues, detected it in nuclei of placental syncytiotrophoblasts and cytoplasm of epithelial cells in prostate tissues. Immunoreactive and catalytically active SULT2B1b was identified in nuclei isolated from term human placenta. Also SULT2B1b was capable of translocating to nuclei in BeWo placental cells after stable transfection and differentiation. In contrast, immunohistochemical analysis of human prostate showed only cytosolic localization of SULT2B1b in the basal and luminal prostate epithelial cells. SULT2B1b was not detected in isolated nuclei from LNCaP prostate cancer cells but was present in the cytosolic fraction. Differential subcellular localization of SULT2B1b in prostate and placenta suggests that SULT2B1b may be differentially regulated and have different physiological functions in these two hormonally responsive human tissues. PMID- 14741046 TI - Stress- and mitogen-induced phosphorylation of the synapse-associated protein SAP90/PSD-95 by activation of SAPK3/p38gamma and ERK1/ERK2. AB - SAPK3 (stress-activated protein kinase-3, also known as p38gamma) is a member of the mitogen-activated protein kinase family; it phosphorylates substrates in response to cellular stress, and has been shown to bind through its C-terminal sequence to the PDZ domain of alpha1-syntrophin. In the present study, we show that SAP90 [(synapse-associated protein 90; also known as PSD-95 (postsynaptic density-95)] is a novel physiological substrate for both SAPK3/p38gamma and the ERK (extracellular-signal-regulated protein kinase). SAPK3/p38gamma binds preferentially to the third PDZ domain of SAP90 and phosphorylates residues Thr287 and Ser290 in vitro, and Ser290 in cells in response to cellular stresses. Phosphorylation of SAP90 is dependent on the binding of SAPK3/p38gamma to the PDZ domain of SAP90. It is not blocked by SB 203580, which inhibits SAPK2a/p38alpha and SAPK2b/p38beta but not SAPK3/p38gamma, or by the ERK pathway inhibitor PD 184352. However, phosphorylation is abolished when cells are treated with a cell permeant Tat fusion peptide that disrupts the interaction of SAPK3/p38gamma with SAP90. ERK2 also phosphorylates SAP90 at Thr287 and Ser290 in vitro, but this does not require PDZ-dependent binding. SAP90 also becomes phosphorylated in response to mitogens, and this phosphorylation is prevented by pretreatment of the cells with PD 184352, but not with SB 203580. In neurons, SAP90 and SAPK3/p38gamma co-localize and they are co-immunoprecipitated from brain synaptic junctional preparations. These results demonstrate that SAP90 is a novel binding partner for SAPK3/p38gamma, a first physiological substrate described for SAPK3/p38gamma and a novel substrate for ERK1/ERK2, and that phosphorylation of SAP90 may play a role in regulating protein-protein interactions at the synapse in response to adverse stress- or mitogen-related stimuli. PMID- 14741048 TI - Structural and functional dissection of the adhesive domains of Plasmodium falciparum thrombospondin-related anonymous protein (TRAP). AB - TRAP (thrombospondin-related anonymous protein) is a sporozoite surface protein that plays a central role in hepatocyte invasion. We have developed procedures for recombinant production of the entire ECD (extracellular domain) and A domain of TRAP using bacterial- and baculovirus-expression systems respectively. The ECD and A domain were purified to homogeneity and migrated on gel-filtration columns as non-aggregated, monomeric proteins. These adhesive modules bound to HepG2 cells in a dose-dependent and bivalent cation-independent manner. The binding of ECD and the A domain to HepG2 cells was inhibited poorly by an excess of sulphatide analogues, suggesting the presence of as yet unidentified receptors for the A domain on hepatocytes. Using surface-plasmon-resonance-based sensor technology (Biacore), we demonstrate that TRAP ECD has higher affinity for heparin (K(D)=40 nM) compared with the A domain (K(D)=79 nM). We also present a three-dimensional structure of the A domain based on the crystal structure of the homologous von Willebrand factor A1 domain. The TRAP A domain shows two spatially distinct ligand-binding surfaces. One surface on the A domain contains the MIDAS (metal-ion-dependent adhesion site) motif, where point mutations of Thr131 and Asp162 correlate with impairment of cell infectivity by sporozoites. The other surface contains a putative heparin-binding site and consists of a basic residue cluster. Our studies suggest that TRAP interacts with multiple receptors during the hepatocyte invasion process. Our results also pave the way for inclusion of these high-quality recombinant TRAP domains in subunit-based vaccines against malaria. PMID- 14741049 TI - A novel neuronal-specific splice variant of Type I phosphatidylinositol 4 phosphate 5-kinase isoform gamma. AB - Type I PIPkins (phosphatidylinositol 4-phosphate 5-kinases) are the enzymes that catalyse the major cellular route of synthesis of PtdIns(4,5) P2, and three isoforms (alpha, beta and gamma) with several splice variants have been found to date. In the present paper, we describe the discovery of a novel splice variant of the gamma isoform, which we call PIPkin Igammac, and which is characterized by the inclusion of a 26-amino-acid insert near the C-terminus. Its transcript appears to be selectively expressed in brain, where it locates in the neurons of restricted regions, such as cerebellum, hippocampus, cortex and olfactory bulb, as indicated by in situ hybridization studies. Overexpression of two different catalytically inactive constructs of PIPkin Igammac in rat cerebellar granule cells causes a progressive loss of their neuronal processes, whereas equivalent kinase-dead versions of PIPkin Igammaa did not induce any such effect, suggesting the possible existence of a specific PtdIns(4,5) P2 pool synthesized by PIPkin Igammac, which is involved in the maintenance of some neuronal cellular processes. PMID- 14741050 TI - A combination of both arginine- and lysine-specific gingipain activity of Porphyromonas gingivalis is necessary for the generation of the micro-oxo bishaem containing pigment from haemoglobin. AB - The black pigment of Porphyromonas gingivalis is composed of the mu-oxo bishaem complex of Fe(III) protoporphyrin IX (mu-oxo oligomer, dimeric haem), namely [Fe(III)PPIX]2O. P. gingivalis W50 and Rgp (Arg-gingipain)- and Kgp (Lys gingipain)-deficient mutants K1A, D7, E8 and W501 [Aduse-Opoku, Davies, Gallagher, Hashim, Evans, Rangarajan, Slaney and Curtis (2000) Microbiology 146, 1933-1940] were grown on horse blood/agar for 14 days and examined for the production of mu-oxo bishaem. Mu-oxo Bishaem was detected by UV-visible, Mossbauer and Raman spectroscopies in wild-type W50 and in the black-pigmented RgpA- and RgpB-deficient mutants (W501 and D7 respectively), whereas no haem species were detected in the straw-coloured colonies of Kgp-deficient strain K1A. The dark brown pigment of the double RgpA/RgpB knockout mutant (E8) was not composed of mu-oxo bishaem, but of a high-spin monomeric Fe(III) protoporphyrin IX species (possibly a haem-albumin complex). In vitro incubation of oxyhaemoglobin with cells of the W50 strain and the RgpA- and RgpB-deficient mutants (W501 and D7) resulted in the formation of mu-oxo bishaem via methaemoglobin as an intermediate. Although the Kgp-deficient strain K1A converted oxyhaemoglobin into methaemoglobin, this was not further degraded into mu-oxo bishaem. The double RgpA/RgpB knockout was also not capable of producing mu-oxo bishaem from oxyhaemoglobin, but instead generated a haemoglobin haemichrome. Inhibition of Arg-X protease activity of W50, W501, D7 and K1A with leupeptin, under conditions where Lys-X protease activity was unaffected, prevented the production of mu-oxo bishaem from oxyhaemoglobin, but resulted in the formation of a haemoglobin haemichrome. These results show that one or both of RgpA and RgpB gingipains, in addition to the lysine-specific gingipain, is necessary for the production of mu-oxo bishaem from haemoglobin by whole cells of P. gingivalis. PMID- 14741051 TI - Calcineurin-mediated Bad translocation regulates cyanide-induced neuronal apoptosis. AB - In cyanide-induced apoptosis, an increase in cytosolic free Ca2+ and generation of reactive oxygen species are initiation stimuli for apoptotic cell death. Previous studies have shown that cyanide-stimulated translocation of Bax (Bcl associated X protein) to mitochondria is linked with release of cytochrome c and subsequent activation of a caspase cascade [Shou, Li, Prabhakaran, Borowitz and Isom (2003) Toxicol. Sci. 75, 99-107]. In the present study, the relationship of the cyanide-induced increase in cytosolic free Ca2+ to activation of Bad ( Bcl 2/Bcl-X(L)- antagonist, causing cell death) was determined in cortical cells. Bad is a Ca2+-sensitive pro-apoptotic Bcl-2 protein, which on activation translocates from cytosol to mitochondria to initiate cytochrome c release. In cultured primary cortical cells, cyanide produced a concentration- and time-dependent translocation of Bad from cytosol to mitochondria. Translocation occurred early in the apoptotic response, since mitochondrial Bad was detected within 1 h of cyanide treatment. Mitochondrial levels of the protein continued to increase up to 12 h post-cyanide exposure. Concurrent with Bad translocation, a Ca2+ sensitive increase in cellular calcineurin activity was observed. Increased cytosolic Ca2+ and calcineurin activation stimulated Bad translocation since BAPTA [bis-(o-aminophenoxy)ethane-N, N, N', N'-tetra-acetic acid], an intracellular Ca2+ chelator, and cyclosporin A, a calcineurin inhibitor, significantly reduced translocation. BAPTA also blocked release of cytochrome c from mitochondria as well as apoptosis. Furthermore, treatment of cells with the calcineurin inhibitors cyclosporin A or FK506 blocked the apoptotic response, linking calcineurin activation and the subsequent translocation of Bad to cell death. These observations show that by inducing a rapid increase in cytosolic free Ca2+, cyanide can partially initiate the apoptotic cascade through a calcineurin-mediated translocation of Bad to mitochondria. PMID- 14741053 TI - Development of a model of focal pneumococcal pneumonia in young rats. AB - BACKGROUND: A recently licensed pneumococcal conjugate vaccine has been shown to be highly effective in the prevention of bacteremia in immunized children but the degree of protection against pneumonia has been difficult to determine. METHODS: We sought to develop a model of Streptococcus pneumoniae pneumonia in Sprague Dawley rats. We challenged three-week old Sprague-Dawley pups via intrapulmonary injection of S. pneumoniae serotypes 3 and 6B. Outcomes included bacteremia, mortality as well histologic sections of the lungs. RESULTS: Pneumonia was reliably produced in animals receiving either 10 or 100 cfu of type 3 pneumococci, with 30% and 50% mortality respectively. Similarly, with type 6B, the likelihood of pneumonia increased with the inoculum, as did the mortality rate. Prophylactic administration of a preparation of high-titered anticapsular antibody prevented the development of type 3 pneumonia and death. CONCLUSION: We propose that this model may be useful for the evaluation of vaccines for the prevention of pneumococcal pneumonia. PMID- 14741052 TI - An increasing number of qualitative research papers in oncology and palliative care: does it mean a thorough development of the methodology of research? AB - BACKGROUND: In the second half of the nineties, a scientific debate about the usefulness of qualitative research in medicine began in the main medical journals as well as the amount of "qualitative" papers published on peer reviewed journals has noticeably increased during these last years. Nevertheless the label of qualitative methodology has been assigned to an heterogeneous collection of studies. Some of them show a complete awareness of the specificity of this kind of research, while others are still largely influenced by the quantitative paradigm prevailing in the medical field. The concern with the rigour and credibility of qualitative methods has lead to the development of a number of checklist for assessing qualitative research. The purposes of this review were to describe the quality of the development of qualitative research in the medical field, focusing on oncology and palliative care, and to discuss the applicability of a descriptive checklist. METHODS: A review was conducted on Medline and PsycINFO databases. On the basis of their abstract, papers found have been classified considering: publication year, kind of journal, paper type, data gathering method, sample size and declared methodological approach. A sub sample of the previous papers was than selected and their methodological characteristics were evaluated based on a descriptive checklist. RESULTS: 351 abstracts and 26 full papers were analysed. An increase over time in the number of qualitative studies is evident. While most of the papers before 1999 were published on nursing journals (43%), afterwards also medical journals were largely represented. Psychological journals increased from 7% to 12%. The 22% of studies used a sample size lower than 15 and the 15% did not specify the sample size in the abstract. The methodological approach was also often not specified and the percentage increased in the second time period (from 73% to 80%). Grounded theory was the most employed methodological approach while phenomenology shows a decrease. Interview remains the most used data gathering method in both periods, even if it shows a 10% reductions, while focus group and multiple methods application both increase to 12%. The use of the descriptive checklist on the full text of the 26 papers shows that all the items present a larger percentage of satisfaction after 1 January 1999 than it was for the paper published before 1999. There seems to be two different types of quality criteria: specific and unspecific. The first ones mainly refer to qualitative paradigm (such as the relationship with the subject of research or evidence about how subjects perceived the research) and they are often not satisfied. In contrast unspecific criteria (such as the connection to an existing body of knowledge or systematic data gathering) which are mainly shared with the quantitative paradigm are more frequently satisfied. CONCLUSIONS: In oncology and palliative care the publication of qualitative studies increased during the nineties, reaching its peak in around 2000. The use of descriptive checklists even if it was not easy to apply, allows researchers to get a deeper insight into methodological facets that a global judgement may leave out. PMID- 14741054 TI - Sensitivity and reproducibility of standardized-competitive RT-PCR for transcript quantification and its comparison with real time RT-PCR. AB - BACKGROUND: Probe based detection assays form the mainstay of transcript quantification. Problems with these assays include varying hybridization efficiencies of the probes used for transcript quantification and the expense involved. We examined the ability of a standardized competitive RT-PCR (StaRT PCR) assay to quantify transcripts of 4 cell cycle associated genes (RB, E2F1, CDKN2A and PCNA) in two cell lines (T24 & LD419) and compared its efficacy with the established Taqman real time quantitative RT-PCR assay. We also assessed the sensitivity, reproducibility and consistency of StaRT PCR. StaRT PCR assay is based on the incorporation of competitive templates (CT) in precisely standardized quantities along with the native template (NT) in a PCR reaction. This enables transcript quantification by comparing the NT and CT band intensities at the end of the PCR amplification. The CT serves as an ideal internal control. The transcript numbers are expressed as copies per million transcripts of a control gene such as beta-actin (ACTB). RESULTS: The NT and CT were amplified at remarkably similar rates throughout the StaRT PCR amplification cycles, and the coefficient of variation was least (<3.8%) when the NT/CT ratio was kept as close to 1:1 as possible. The variability between the rates of amplification in different tubes subjected to the same StaRT PCR reaction was very low and within the range of experimental noise. Further, StaRT PCR was sensitive enough to detect variations as low as 10% in endogenous actin transcript quantity (p < 0.01 by the paired student's t-test). StaRT PCR correlated well with Taqman real time RT-PCR assay in terms of transcript quantification efficacy (p < 0.01 for all 4 genes by the Spearman Rank correlation method) and the ability to discriminate between cell types and confluence patterns. CONCLUSION: StaRT PCR is thus a reliable and sensitive technique that can be applied to medium-high throughput quantitative transcript measurement. Further, it correlates well with Taqman real time PCR in terms of quantitative and discriminatory ability. This label-free, inexpensive technique may provide the ability to generate prognostically important molecular signatures unique to individual tumors and may enable identification of novel therapeutic targets. PMID- 14741055 TI - "Closing-in" phenomenon in Alzheimer's disease and subcortical vascular dementia. AB - BACKGROUND: The 'closing-in' phenomenon is defined as a tendency to close in on a model while copying it. This is one of several constructional apraxia observed in dementia, particularly in Alzheimer's disease (AD). The aim of this study was to investigate the usefulness of it in the differential diagnosis of AD and subcortical vascular dementia (SVD) and to clarify the factors associated with it. METHODS: We operationally defined and classified it into three types, namely overlap, adherent, and near type. We analyzed the incidence of it in patients with AD (n = 98) and SVD (n = 48). RESULTS: AD patients exhibited a significantly higher occurrence of it as compared to SVD patients. Among the different types of it, the overlap and adherent types occurred almost exclusively in AD patients. A discriminant analysis in AD subjects revealed that the scores obtained from the MMSE, CDR, Barthel index, and the Rey-Osterrieth complex figure test were correlated significantly with the occurrence of it. There was no statistical difference between the Q-EEG parameters of patients that exhibited the closing-in phenomenon and those that did not. CONCLUSIONS: This study suggests that the closing-in phenomenon is phase- and AD-specific and might be a useful tool for the differential diagnosis of AD and SVD. PMID- 14741056 TI - Insulin receptor activation and down-regulation by cationic lipid transfection reagents. AB - BACKGROUND: Transfection agents comprised of cationic lipid preparations are widely used to transfect cell lines in culture with specific recombinant complementary DNA molecules. We have found that cells in culture are often resistant to stimulation with insulin subsequent to treatment with transfection agents such as LipofectAMINE 2000 and FuGENE-6. This is seen with a variety of different readouts, including insulin receptor signalling, glucose uptake into muscle cells, phosphorylation of protein kinase B and reporter gene activity in a variety of different cell types RESULTS: We now show that this is due in part to the fact that cationic lipid agents activate the insulin receptor fully during typical transfection experiments, which is then down-regulated. In attempts to circumvent this problem, we investigated the effects of increasing concentrations of LipofectAMINE 2000 on insulin receptor phosphorylation in Chinese hamster ovary cells expressing the human insulin receptor. In addition, the efficiency of transfection that is supported by the same concentrations of transfection reagent was studied by using a green fluorescent protein construct. Our data indicate that considerably lower concentrations of LipofectAMINE 2000 can be used than are recommended by the manufacturers. This is without sacrificing transfection efficiency markedly and avoids the problem of reducing insulin receptor expression in the cells. CONCLUSION: Widely-used cationic lipid transfection reagents cause a state of insulin unresponsiveness in cells in culture due to fully activating and subsequently reducing the expression of the receptor in cells. This phenomenon can be avoided by reducing the concentration of reagent used in the transfection process. PMID- 14741057 TI - Chronic rTMS induces subsensitivity of post-synaptic 5-HT1A receptors in rat hypothalamus. AB - Chronic administration of several antidepressants, notably the selective serotonin re-uptake inhibitors (SSRIs) induces sub-sensitivity of post-synaptic 5 HT1A receptors in the hypothalamus. Chronic repetitive transcranial magnetic stimulation (rTMS) is a form of treatment for depression which is often compared to electroconvulsive shock therapy (ECT). rTMS was applied to rats either on a single occasion (acute) or daily for 8 d (chronic). Twenty-four hours after the last treatment, the rats were injected with saline or 8-OH-DPAT (50 microg/kg). The rats were killed 20 min later and trunk blood taken for measurement of corticosterone and ACTH levels. Chronic rTMS did not affect basal corticosterone or ACTH levels but significantly blunted the responses to 8-OH-DPAT, while acute rTMS had no effect on either basal or 8-OH-DPAT-stimulated responses. In common with several other antidepressant treatments, chronic rTMS reduces the sensitivity of post-synaptic 5-HT1A receptors in the hypothalamus. This effect may be significant in relation to the therapeutic mechanism of rTMS. PMID- 14741058 TI - Decreased expression of mineralocorticoid receptor mRNA in the prefrontal cortex in schizophrenia and bipolar disorder. AB - Abnormal prefrontal cortical activity, activation of the hypothalamic-pituitary adrenal (HPA) axis, and deficits in slow-wave sleep (SWS) have been extensively reported in patients with affective disorders and schizophrenia, yet the underlying pathophysiological mechanisms have not been completely elucidated. Mineralocorticoid receptor (MR) and glucocorticoid receptor (GR) are two nuclear hormone receptors of primary importance in the control of stress-related and circadian HPA activity. A recent study showed that blocking brain MR activity not only enhances CRF-induced ACTH and cortisol release, but also significantly reduces SWS in humans. We hypothesized that the expression of MR would be deficient in the prefrontal cortex of patients with schizophrenia and affective disorders. The MR mRNA expression in the post-mortem prefrontal cortex of patients with major depression (MD), bipolar (BP), and schizophrenic (SZ) disorders and non-psychiatric controls (n=15 for each patient group, and n=14 for controls) was determined by in-situ hybridization. In the dorsolateral prefrontal cortex Brodmann's area 9 (BA 9), MR mRNA was significantly lower (p<0.05) in all laminae (I-VI) in BP, and in laminae I, III, IV and VI in SZ than in the controls. MR mRNA in BA 9 was negatively correlated with the duration of psychiatric illnesses. In BA 46, MR mRNA was not significantly different among groups, but was positively correlated with brain pH. These results provide the first evidence of deficient prefrontal MR mRNA expression in BP and SZ. Whether these findings may be linked to the abnormal prefrontal function, HPA axis activation, or the deficits in SWS found in these major psychiatric illnesses remains to be further explored. PMID- 14741059 TI - Revising polypharmacy to a single antipsychotic regimen for patients with chronic schizophrenia. AB - Antipsychotic polypharmacy has been empirically used and a recent trend in favour of that mode of therapy has been suggested for the treatment of schizophrenia. The clinical efficacy, however, still remains to be clarified. In order to critically evaluate the usefulness of such kind of psychopharmacotherapy, antipsychotic combination regimen (polypharmacy) was switched to a treatment with the single main antipsychotic (monotherapy) in cross-tapered fashion, while approximately maintaining the total amount, for patients with chronic schizophrenia. Patients had been treated with an average of three antipsychotics and maintained with the same antipsychotic polypharmacy regimen for more than 6 months before the entry. They were followed up with an antipsychotic monopharmacy and evaluated at 24 wk after completion of switching. Forty-seven patients were recruited for this study. Of 44 patients for whom evaluation was possible, 24 (54.5%) remained stable, while 10 (22.7%) showed improvement and the same number of patients ended in a deleterious status. Twenty-two patients were converted to antipsychotic monotherapy, while another 12 needed minimal dosing of low-potency agents. Overall, social functioning, evaluated by the Global Assessment of Functioning and the Clinical Global Impression, remained unchanged. Eighteen of 34 successful patients showed adverse effects of the main antipsychotic medication, which necessitated a significant dose reduction. Nine out of 10 deteriorating patients had been treated with a combination of low- and high potency antipsychotics. It is suggested that many instances of antipsychotic polypharmacy is avoidable. The result is compatible with the current treatment recommendations, which dictate the use of a single antipsychotic agent. PMID- 14741060 TI - A double-blind, placebo controlled, cross-over trial of adjunctive donepezil for cognitive impairment in schizophrenia. AB - Although there have been several case reports suggesting the beneficial effect of acetylcholinesterase inhibitors in the cognitive deficits seen in schizophrenia, controlled studies have revealed contradictory results. The aim of this study was to investigate if donepezil could improve cognitive functions in schizophrenia. Twelve schizophrenic patients, who were diagnosed according to DSM-IV criteria and who had been on a stable dose of a high-potency typical antipsychotic for a minimum period of 3 months, participated in this 12-wk double-blind, placebo controlled, cross-over study of donepezil adjunctive treatment. Patients were randomly assigned under double-blind conditions to receive 5 mg/d donepezil or placebo for 6 wk, and then were crossed over to the alternate condition for 6 additional weeks. At baseline, 6 and 12 wk, patients were evaluated with the Positive and Negative Syndrome Scale (PANSS), Calgary Depression Scale, the Wechsler Memory Scale Revised (WMS-R), a test for Verbal Fluency, Trail Making Test, Parts A and B, and Wisconsin Card Sorting Test (WCST). Treatment effect was not significant in any of the cognitive measures. There were also no significant changes in the PANSS and depression scores. Nicotinic receptor desensitization may cause non-responsiveness to acetylcholine as previously suggested, but the most likely explanation appears to be that defects in other neurotransmitter systems account for the cognitive deficits seen in schizophrenic patients. PMID- 14741061 TI - Protective effect of 1-methyl-1,2,3,4-tetrahydroisoquinoline against dopaminergic neurodegeneration in the extrapyramidal structures produced by intracerebral injection of rotenone. AB - The aim of this paper was to investigate whether rotenone, a pesticide causing experimental parkinsonism, causes direct damage to dopaminergic structure when injected intracerebrally and whether this action may be prevented by peripheral administration of 1-methyl-1,2,3,4-tetrahydroisoquinoline (1MeTIQ), an endogenous compound with anti-dopaminergic activity. Male Wistar rats were injected unilaterally into the median forebrain bundle with 2 microg rotenone, and received 1MeTIQ, 50 mg/kg i.p. 1 h before and then daily for 21 d. To compare the effect of intracerebral and peripheral treatment, rotenone was also given once or for 7 d in a dose of 10 mg/kg s.c. Dopamine, serotonin and their metabolites were assessed by HPLC in the substantia nigra and striatum. While a single subcutaneous rotenone dose did not produce any change in striatal dopamine metabolism, the multiple treatments resulted in changes suggesting a shift in the metabolism towards oxidative desamination and reduction of O-methylation. In contrast to systemic injections, intracerebral-administered rotenone produced a decrease in dopamine and its metabolites content in the striatum (dopamine decrease by 70%) and substantia nigra (dopamine decrease by 35%), without affecting the serotonin system. As those changes were observed 21 d after the injection of rotenone, they suggest a durable neurotoxic effect. The treatment with 1MeTIQ strongly reduced the fall of striatal dopamine concentration. The data suggest that rotenone given peripherally affects metabolic processes in dopaminergic neurons, and this seems to result from its neurotoxic action, which may be observed after an intracerebral injection. 1MeTIQ is able to counteract the damaging action of rotenone and seems to be a potential neuroprotective agent. PMID- 14741062 TI - Switching elderly chronic psychotic patients to olanzapine. AB - The purpose of this study was to examine whether elderly chronic schizophrenia or schizoaffective disorder patients would clinically improve if switched to olanzapine from previous neuroleptic treatment. Twenty-one hospitalized patients, aged 6088 yr, with a diagnosis of chronic schizophrenia or schizoaffective disorder who were being treated with typical neuroleptic medication were switched to olanzapine. The Positive and Negative Symptom Scale (PANSS), Geriatric Depression Scale (GDS) and Clinical Global Impression Severity (CGI-S) Scale were completed while patients were on their previous medication regimen and again 6 months after the last patient had been started on olanzapine. The mean duration of treatment was 289 d (S.D.=139). Three patients discontinued the medication. Mean end dose of olanzapine was 12.9 mg (S.D.=5.7). Paired sample t tests were used to test change on PANSS Positive, Negative and Total scales, CGI, GDS and body weight. PANSS (Positive, p=0.002; Negative, p=0.003; General, p=0.003; and Total, p=0.000) and CGI (p=0.000) but not the GDS (p=0.67) demonstrated statistically significant improvement. There was no significant change in body weight (p=0.61). Elderly patients with aggravation of chronic schizophrenia showed improvement after being switched to olanzapine with no weight gain. Clinically meaningful change was observed in positive and negative psychotic symptomatology but not in depressive symptoms. PMID- 14741063 TI - Citalopram challenge in social anxiety disorder. AB - The aim of the present study was to evaluate neuroendocrine and behavioural responses to a challenge with the selective serotonin (5-HT) reuptake inhibitor citalopram (Cit) in patients with social anxiety disorder (SAD). Cit was given intravenously (20 mg over 30 min) to 18 patients with SAD and 18 matched healthy subjects in a double-blind, placebo-controlled cross-over design. Cit challenge resulted in the increased plasma concentrations of cortisol and prolactin relatively to placebo without significant differences between the patients and controls. The patients had higher ratings of anxiety that were not affected by Cit, and more headaches than controls after Cit. Thus, the neuroendocrine sensitivity to 5-HT stimulation with Cit in patients with SAD was not different from the response in controls indicating lack of major alterations in the function of 5-HT receptors. The increased headache in patients may suggest hypersensitivity of some subtypes of 5-HT receptors in SAD. PMID- 14741064 TI - Down-regulation of nitric oxide synthase in the diabetic rabbit kidney: potential relevance to the early pathogenesis of diabetic nephropathy. AB - OBJECTIVES: Nephropathy is a well-recognised complication of diabetes mellitus (DM). The aim of this study was to investigate the effect of DM on the density and distribution of nitric oxide (NO) synthase (NOS) in the rabbit kidney. Quantification of the NOS radioligand on slide-mounted sections was compared with the nitroblue tetrazolium reaction, where the intensity of the reaction varies with the nicotinamide adenine dinucleotide diaphorase (NADPH-d) activity of NOS. MATERIALS AND METHODS: DM was induced with alloxan in six New Zealand White (NZW) rabbits. Plasma creatinine, urea and electrolytes were monitored at monthly intervals. The kidneys were removed following 6 months of DM. Transverse serial sections were cut and low-resolution autoradiography was performed using a radioligand for NOS ([(3)H]-NOARG). Histochemical localisation of NADPH-d activity was also performed. Densitometric analysis was performed on the autoradiographs and the results compared with those obtained from six age-matched control rabbits. RESULTS: There was a significant (p < 0.01) rise in plasma creatinine levels in the diabetic rabbits, although the mean values remained within the reference range. There was a significant (p < 0.0001) down-regulation of NOS binding sites in both the cortex and medulla of the DM kidney when compared with the controls. A similar decrease in NADPH-d activity was seen in the diabetic renal cortex and medulla. In addition, NADPH-d activity also appeared to be reduced in the diabetic glomeruli when compared with controls. CONCLUSIONS: NOS binding sites and NADPH-d activity are significantly decreased in the DM renal cortex and medulla. These changes are associated with a mild deterioration in renal function and may be an early event that could subsequently play a role in the progression of DM nephropathy. Manipulating the NO pathway during the early stages of DM nephropathy may be beneficial. PMID- 14741065 TI - Epidural block with ropivacaine and bupivacaine for elective caesarean section: maternal cardiovascular parameters, comfort and neonatal well-being. AB - OBJECTIVE: To determine cardiovascular effects and neonatal outcome of ropivacaine 0.75% and bupivacaine 0.5% for elective epidural caesarean section. RESEARCH DESIGN AND METHODS: Healthy pregnant women, scheduled for elective caesarean section, were enrolled in this randomised, double-blind study. Epidural block was obtained with 20-30 ml of ropivacaine 0.75% (Group R) or bupivacaine 0.5% (Group B) and surgery did not commence until anaesthesia was achieved bilaterally to T6. MAIN OUTCOME MEASURES: Maternal heart rate and blood pressure were assessed before the main dose of local anaesthetic and at 5-min intervals until 35 min. Neonatal umbilical pH and Apgar scores were determined after delivery. Ten, twenty and thirty minutes after the main dose, sensory and motor block characteristics were determined. Quality of analgesia was assessed by the anaesthetist, surgeon and the patient. Adverse events were recorded. RESULTS: Sixty-two patients were enrolled and the data of 60 of them were eligible for analysis: 31 in Group R and 29 in Group B. The area under the curve (AUC) for maternal heart rate decreased significantly less in Group B than in Group R (p = 0.038). Twenty-five and thirty minutes after administration of the main local anaesthetic dose, heart rate decreased significantly less in Group B than in Group R (p = 0.006 and p = 0.007). There was no difference in AUC for maternal blood pressure (p = 0.32). Repeated measurement analysis showed no difference between groups in motor block (p = 0.78) and in spread of the sensory block (lower level: p = 0.83, upper level: p = 0.88). There was no statistical difference in neonatal umbilical pH (p = 0.22) and Apgar score (p = 0.59). Multiple linear regression analysis showed a significant influence of maternal body mass index on neonatal pH (p = 0.004), but not of maternal blood pressure (p = 0.323), nor of maternal heart rate (p = 0.12). The quality of analgesia and incidence of adverse events were similar in both groups. CONCLUSIONS: Both drugs produced equally satisfactory epidural block. Although ropivacaine 0.75% resulted in a greater decrease of maternal heart rate, this effect did not influence neonatal well-being. Both ropivacaine 0.75% and bupivacaine 0.5% can therefore be recommended for epidural anaesthesia in elective caesarean section. PMID- 14741066 TI - Efficacy of once-daily extended-release lovastatin as compared to immediate release lovastatin in patients with hypercholesterolemia. AB - OBJECTIVE: to compare the efficacy and safety of 20 mg of lovastatin when administered once daily as an extended-release (ER) tablet or as an immediate release (IR) tablet. RESEARCH DESIGN AND METHODS: Male or female patients aged 21 70 years with hypercholesterolemia who provided written informed consent and met the inclusion criteria were screened. A total of 179 patients were enrolled: 100 male and 79 female; 153 were Caucasian, eight Black and 18 other races; the mean age was 56 years. Patients were generally in good health as evidenced by medical history, physical and laboratory examination. Patients were required to not exceed specific low-density lipoprotein cholesterol (LDL-C) levels depending on their risk category. The trial was conducted as a multi-center, randomized, double-blind, positive-controlled, double-dummy, two-way crossover study. Patients were washed-out of any prior lipid-lowering medications (period 1) and then received one ER or one IR lovastatin tablet for 12 weeks (period 2) and then washed out with placebo for 6 weeks (period 3). They then received the alternate treatment for an additional 12 weeks (period 4). MAIN OUTCOME MEASURES: The primary efficacy variable was the combined mean percent change in LDL-C from baseline to endpoint for periods 2 and 4. Secondary variables included the mean percent change from baseline in high-density lipoprotein cholesterol (HDL-C), total cholesterol (TC), and triglycerides (TG) for periods 2 and 4 combined. Least-square mean differences between ER and IR treated groups were estimated at both treatments. All tests were two-sided and a p-value of < 0.05 was considered statistically significant. RESULTS: Both ER and IR lovastatin tablets produced statistically significant changes in the lipid profile from baseline. Differences in HDL-C (4.1% and 4.3% for ER and IR, respectively) and TG (7.4% and 10.4% for ER and IR, respectively) were not significant between treatments. TC (19.1% and 17.2% for ER and IR, respectively) and LDL-C (26.4% and 23.1% for ER and IR, respectively) were also reduced significantly from baseline by both treatments. However the ER lovastatin reduced TC by an additional 1.9% (p = 0.0355) and LDL-C by a further 3.3% decrease (p = 0.0028) as compared to the IR formulation. The increase in LDL-C efficacy is equivalent to an increase of 50% in the dose of IR lovastatin, suggesting that 20 mg ER is equivalent to about 30 mg IR in LDL-C lowering capacity. No apparent difference in the safety profile between the two formulations was noted. CONCLUSIONS: The data show that 20 mg of ER lovastatin was about one and one-half times as effective at lowering LDL-C than the same dose of IR lovastatin. Both regimens were tolerated well. PMID- 14741067 TI - Effect of interleukin 8 and ICAM-1 on calcium-dependent outflow of K+ in erythrocytes from subjects with essential hypertension. AB - INTRODUCTION: The pathogenic mechanisms underlying the increase in peripheral resistance and the contraction of smooth muscular fibre cells in essential hypertension are not yet clearly understood. However, it is now known that immune system activation plays a role in the pathogenesis of some forms of arterial hypertension, and recent data show that the Ca2+ influx in some cells (i.e. red blood cells, leukocytes, platelets, smooth muscular fibre cells) is increased in subjects with essential hypertension, thus revealing a possible alteration in cellular membrane. The end-points of this study were therefore to ascertain whether red blood cells used as a cellular membrane model have a greater Ca2+ dependent K+ flow (Gardos effect) in hypertensive patients than in normotensive controls, to point out a different regulation of ionic channels, and whether IL-8 and the adhesion molecule ICAM-1 influence the membranous outflow. MATERIAL AND METHODS: The study was conducted on 87 Caucasian subjects. Of these, 50 (25 men, 25 women; mean age 43 +/- 3 years, mean body mass index (BMI) 27 +/- 0.5 and 22.3 +/- 0.3 kg/m(2), respectively) had mild-to-moderate hypertension (mean arterial blood pressure 120 +/- 8 mmHg ). The other 37 (18 men, 19 women; mean age 39 +/- 3 years; BMI 23.8 +/- 0.5 kg/m(2) and 22.8 +/- 0.5 kg/m(2), respectively were normotensive healthy volunteers (mean arterial blood pressure 89 +/- 2 mmHg). All the patients and subjects were untreated for at least 4 weeks before blood sampling. RESULTS: Ca2+-dependent K+ outflow was found to be greater in samples from patients with essential hypertension than in those from normotensive controls. lL-8 and ICAM-1 significantly enhanced the Ca2+-dependent K+ outflow in red blood cells from hypertensive subjects but had an inhibitory effect on cells from controls. In the experimental model, the presence of TMB-8, a membrane calcium antagonist, significantly reduced the Ca2+-dependent K+ efflux. CONCLUSION: Vasoconstriction in subjects with essential hypertension may therefore depend on a different regulation of ionic flow that probably supports an increased Ca2+ inflow in smooth muscle fibre cells. Under certain pathological conditions, some immune system components (i.e. interleukins, adhesion molecules) may directly enhance membrane permeability to Ca2+, thus inducing vasoconstriction in the smooth muscle cells. PMID- 14741068 TI - Rationale and design of the prospective and retrospective observational study of Avonex and Rebif (PROOF) for the treatment of relapsing-remitting multiple sclerosis. AB - Previous studies comparing the two available interferon beta (IFNbeta)-1a products, Avonex and Rebif, for the treatment of relapsing-remitting multiple sclerosis (RRMS) have been limited and of short duration. Therefore, the Prospective and Retrospective Observational Study of Avonex and Rebif (PROOF) was designed to provide long-term (up to 5 years) comparative data on the efficacy, safety, and tolerability of these two agents. Patients with RRMS receiving treatment with either Avonex 30 microg intramuscularly once weekly or Rebif 44 microg subcutaneously three times weekly from 12 to 24 months are being enrolled and will continue their respective treatments for the 36-month duration of the study. The primary efficacy endpoint will be change in brain parenchymal fraction, which will be evaluated through magnetic resonance imaging scans by blinded radiologists. Secondary endpoints will include the following: relapse rates; intravenous steroid use; the proportion of patients with an increase of > or = 1 point on the Expanded Disability Status Scale (EDSS) and with an increase in EDSS score sustained for 6 months; mean change in EDSS score; cumulative number of new or enlarging T2 lesions; T2 and T1 lesion volumes; gadolinium enhanced lesion number and volume; and safety and tolerability. The study design of PROOF will permit more meaningful conclusions regarding the optimal IFNbeta-1a product for the long-term treatment of patients with multiple sclerosis. PMID- 14741069 TI - Insulin glargine: long-acting basal insulin analog for improved metabolic control. AB - The primary aim of insulin therapy is to replace endogenous insulin secretion in patients with type 1 or type 2 diabetes in a physiologically sound manner, mimicking normal secretion patterns to adequately regulate glucose metabolism. The currently available human insulins for basal therapy--neutral protamine Hagedorn (NPH), Lente and Ultralente--and analogs such as insulin glargine, differ in pharmacokinetic properties. Clinical trial data indicate that insulin glargine may satisfy basal insulin requirements, with an improved safety profile relative to other available insulins used for basal supplementation. This review describes the unique pharmacokinetic properties and clinical efficacy of insulin glargine. PMID- 14741070 TI - TNF-alpha inhibition for potential therapeutic modulation of SARS coronavirus infection. PMID- 14741071 TI - The effect of nateglinide on the pharmacokinetics and pharmacodynamics of acenocoumarol. AB - OBJECTIVE: The potential for a drug interaction was investigated between nateglinide, an oral antidiabetic agent, and acenocoumarol, an oral anticoagulant, as these drugs are primarily metabolized via CYP2C9. METHODS: A two-period, randomized, double-blind, two-way crossover study design was employed to evaluate the effect of nateglinide on the pharmacokinetics and pharmacodynamics of acenocoumarol in 11 healthy male or female subjects. All subjects received either nateglinide 120 mg t.i.d. or placebo for 5 days in a crossover fashion and a single 10-mg dose of acenocoumarol on day 3. Plasma concentrations of R- and S-acenocoumarol and the anticoagulation parameters [prothrombin time (PT) and international normalized ratio of PT (PTINR)] were determined for 72 h following acenocoumarol administration. The pharmacokinetic and pharmacodynamic parameters of acenocoumarol were determined by noncompartmental analysis. RESULTS: The mean (coefficient of variation (CV%)) area under the concentration-time curve (AUC(0-t)) of R-acenocoumarol in the presence and absence of nateglinide was 4217 (23%) and 3831 (24%) ng.h/ml, respectively. The corresponding values for S-acenocoumarol were 397 (20%) and 382 (23%), respectively. The mean (CV%) C(max) of R-acenocoumarol in the presence and absence of nateglinide was 304 (16%) and 316 (16%), respectively and the corresponding values for S-acenocoumarol were 142 (36%) and 141 (34%), respectively. The 90% confidence intervals indicated that exposure parameters, AUC(0-t) and C(max), of both R- and S-acenocoumarol were within the acceptable limits of 0.8-1.25. The mean (CV%) of area under the concentration-time curve of PT (AUC(PT)) following acenocoumarol administration in the presence and absence of nateglinide was 1170 (10%) and 1136 (8%), respectively. The corresponding AUC(INR) values were 104 (13%) and 99 (10%), respectively. Nateglinide co administration has no influence on the PT or PTINR of acenocoumarol (p > 0.05). CONCLUSION: Co-administration of nateglinide does not influence either the pharmacokinetics or the anticoagulant activity of R- and S-acenocoumarol in healthy subjects. This suggests that no dosage adjustments will be required when nateglinide and acenocoumarol are coadministered in clinical practice. PMID- 14741072 TI - Effect of preventive treatment with botulinum toxin type A on acute headache medication usage in migraine patients. AB - OBJECTIVE: To evaluate the impact of preventive treatment of migraine with botulinum toxin type A (BoNT-A as BOTOX) on the amount of acute headache medications used. RESEARCH DESIGN AND METHODS: Data from four studies of BoNT-A treatment for migraine were pooled for an aggregate analysis. All studies were at least 12 weeks in duration. For each study, the amounts of headache medications used at weeks 8-12 following BoNT-A treatment were compared with pretreatment baseline amounts and expressed preventive headache care considering acute as a percentage change. MAIN OUTCOME MEASURES: The mean value for the reduction in medication usage was calculated by pooling data from the individual studies and weighting the data according to the sample size of each study. RESULTS: Four studies (one published, and three presented as recent meeting abstracts) with a total of 167 patients quantified acute headache medication use before and after BoNT-A treatment. The weighted average reduction in medication usage (primarily triptans) was 57% (range 38-75%). CONCLUSIONS: The results of this pooled analysis indicated a 57% reduction in acute headache medication use in the 8- to 12-week period following injection of BoNT-A. A reduction of this magnitude could represent substantial savings in the costs of acute medications. This could help to offset the total cost of treatment and suggests BoNT-A may be a cost reasonable option for medication offsets alone especially in patients with chronic headache with higher acute medication use. Additional larger controlled efficacy and safety studies must be done to confirm these results since three of the four studies were preliminary research and of different study types. Further prospective studies of direct and indirect costs, including those for disability and lost productivity, are needed to evaluate the overall impact of BoNT-A therapy on the economic, societal, and individual burden of migraine headache. PMID- 14741073 TI - Waist-hip ratio and low HDL predict the risk of coronary artery disease in Pakistanis. AB - OBJECTIVE: To establish risk factor causal associations for coronary artery disease (CAD) in the native Pakistani population. METHODS: We conducted a hospital-based, case-control study of 200 cases with angiographically documented CAD and 200 age- and sex-matched controls without angiographic evidence of CAD. Patients on lipid lowering therapy were excluded. Lifestyle, anthropometric and biochemical risk factors were assessed in both groups. RESULTS: The presence of CAD was associated with current, past or passive smoking, a history of diabetes and high blood pressure, a positive family risk factors in this study; levels were below history of CAD, body fat percentage, waist-hip ratio (WHR), low apolipoprotein A1 or low HDL, lipoprotein (a), glucose, insulin, insulin resistance, C-reactive protein (CRP), total cholesterol to HDL ratio (TC/HDL) and creatinine on univariate conditional logistic regression analysis. In multiple regression analysis, significant independent associations were found with low HDL (OR 0.11; 95% CI 0.04-0.34; p < 0.001) positive family history (OR 1.79; 95% CI 1.09-2.93; p = 0.02), CRP (OR 1.45; 95% CI 1.19-1.75; p < 0.001) and WHR (OR 1.04; 95% CI 1.01-1.08; p = 0.01). Angiograms were also quantified for the extent and severity of CAD by the Gensini scoring system. Quantitative angiographic data showed associations with age (p = 0.01), the duration of diabetes (p = 0.04), WHR (p = 0.06), low HDL (p < 0.001), lipoprotein (a) (p = 0.001), creatinine (p < 0.001) and CRP (p = 0.007). Results indicate that total and LDL cholesterol were not significant currently accepted thresholds for treatment. CONCLUSIONS: The cardiovascular risk profile in this population is consistent with metabolic syndrome where low HDL and WHR can be used to predict the risk of CAD. Results suggest the need to redefine the currently practised approach to CAD management in this population to fit local needs. PMID- 14741074 TI - Double-blind, randomised, placebo-controlled study to evaluate the efficacy and safety of a fixed combination containing two plant extracts (Crataegus oxyacantha and Eschscholtzia californica) and magnesium in mild-to-moderate anxiety disorders. AB - OBJECTIVE: To assess the clinical efficacy of a neurotonic component containing fixed quantities of two plant extracts (Crataegus oxyacantha and Eschscholtzia californica) and magnesium versus placebo in mild-to-moderate anxiety disorders with associated functional disturbances, under usual general practice prescription conditions. RESEARCH DESIGN AND METHODS: A total of 264 patients (81% female; mean age: 44.6 years) presenting with generalised anxiety (DSM-III R) of mild-to-moderate intensity (total Hamilton anxiety scale score between 16 and 28) were included in a double-blind, randomised, placebo-controlled trial. Patients were randomly assigned to two groups: 130 received the study drug (Sympathyl), and 134 a placebo (two tablets twice daily for 3 months). Efficacy and safety data were recorded before first administration and 7, 14, 30, 60 and 90 days after start of treatment. MAIN OUTCOME MEASURES: Efficacy was assessed by (a) change in Hamilton anxiety scale total and somatic scores; (b) change in patient self-assessment; (c) number and percentage of responsive subjects (reduction of at least 50% in Hamilton or self-assessment score); and (d) the physician's clinical global impression. Tolerance was assessed by undesirable events spontaneously reported by the patients over the study period. RESULTS: Total and somatic Hamilton scale scores and subjective patient-rated anxiety fell during treatment, indicating clinical improvement. The decrease was greater in the study drug than in the placebo group. End of treatment clinical improvement, as measured by the mean difference between final and pre-treatment scores, was, for the study drug and placebo groups respectively: -10.6 and -8.9 on the total anxiety score (p = 0.005); -6.5 and -5.7 on the somatic score (p = 0.054); and 38.5 and -29.2 for subjectively assessed anxiety (p = 0.005). The risk/benefit ratio as judged by the investigating physicians was also significantly greater in the study drug than in the placebo group. In all, 15 patients (11.5%) in the study drug group and 13 patients (9.7%) in the placebo group experienced 22 and 15 adverse events, respectively. Undesirable events were mainly mild or moderate digestive or psychopathological disorders. CONCLUSIONS: The preparation containing fixed quantities of Crataegus oxyacantha, Eschscholtzia californica, and magnesium proved safe and more effective than placebo in treating mild-to moderate anxiety disorders. Sympathyl is produced and marketed by Laboratoire Innotech International, Arcueil, France. PMID- 14741075 TI - Anti-inflammatory activity of H1-receptor antagonists: review of recent experimental research. AB - OBJECTIVE: To compare the anti-inflammatory effects of fexofenadine with other H(1)-receptor antagonists in vitro. DATA SOURCES: Published literature. STUDY SELECTION: Recent experimental studies on anti-inflammatory effects of H(1) receptor antagonists. Databases searched: Medline, Medscape. PERIOD COVERED: 1990 2003. Search terms: second-, third-generation antihistamines; sedating, nonsedating antihistamines; in vitro anti-inflammatory activity; cetirizine; ebastine; loratadine; fexofenadine; desloratadine. RESULTS: Second- and third generation H(1)-receptor antagonists may demonstrate significant in vitro anti inflammatory activity at concentrations considered to be clinically relevant. In some instances, higher (supraclinical) concentrations are required to achieve comparable effects. CONCLUSIONS: Experimental research suggests that second- and third-generation H(1)-receptor antagonists may achieve anti-inflammatory effects in a clinical context. Further studies are required to support this conclusion. PMID- 14741076 TI - Pure red cell aplasia with anti-erythropoietin antibodies occurs more commonly with one formulation of epoetin alfa than another. AB - OBJECTIVE: To compare rates of pure red cell aplasia (PRCA) over time in patients with chronic renal failure treated with subcutaneous injections of two brands of epoetin alfa (either Eprex or Epogen) or epoetin beta (NeoRecormon). METHODS: Cases of antibody-mediated PRCA associated with epoetin alfa-treated patients were obtained from public databases and company websites and limited to time periods when exposure data also were available The subcutaneous exposure rates per 100 000 patient-years were calculated for the periods 1989-1998 and 1999 2002. RESULTS: The event rate for antibody-mediated PRCA for Epogen and Eprex were similar from 1989 to 1998, but the number of cases of Eprex-associated PRCA has increased markedly since 1999, even after accounting for subcutaneous exposure. In contrast, rates have remained low for Epogen and NeoRecormon. CONCLUSIONS: The recent increase in PRCA appears to be product specific and cannot be explained solely as a consequence of increased use of the subcutaneous route of administration. PMID- 14741077 TI - SCID repopulating cells derived from unmobilised adult human peripheral blood. AB - Severe combined immunodeficient (SCID)-repopulating cells (termed SRC) with lymphohaematopoietic differentiation potential reside at an extremely low frequency in unmobilised adult human peripheral blood. Recently, an ex vivo method of increasing the relative numbers of at least four distinct human stem cell classes, that include CD34+ haematopoietic progenitor cells, in mononuclear cells (MNC) obtained from unmobilised adult human peripheral blood has been described. This process is triggered by a monoclonal antibody (mAb) against the human monomorphic region of the beta chain of HLA-DP, DQ and DR (clone CR3/43). Herein, we assess the ability of human male donor-derived MNC, following ex vivo culturing for 3 hr in haematopoietic-conducive conditions (HCC) (3-hr MNC/HCC), to form SRC in female non-obese diabetic/severe combined immunodeficient (NOD/SCID) mice. All 3-hr MNC/HCC-recipient animals exhibited significant levels (> 0.5%) of human cell engraftment in the bone marrow, thymus and spleen when compared to animals receiving MNC cultured in the absence of CR3/43. Phenotypic characterisation of the bone marrow cell populations of engrafted mice demonstrated significant levels of human lymphohaematopoietic cell lineages, comprised of T lymphocytes, monocytes, erythrocytes and megakaryocytes, including platelets. In addition, significant levels of clonogenic human CD34+ cells were also detected by in vitro surrogate assay. The thymi of engrafted animals contained maturating human thymocytes, while the spleen consisted mainly of T lymphocytes. Fluorescence in situ hybridisation (FISH) further identified the presence of human male X and Y chromosomes at engrafted sites, whilst the human origin of the cells was confirmed by a specific PCR assay for the human Cart-1 gene. In conclusion, the conversion of MNC to SRC in response to treatment with CR3/43 for 3 hr could have far-reaching clinical implications especially where time and donor-histocompatibility are limiting factors. PMID- 14741078 TI - Post-traumatic stress disorder in UK police officers. AB - Prevalence rates for post-traumatic stress disorder (PTSD) in police officers may be six or more times the prevalence rates for the community. Once established, is PTSD in police officers more severe than PTSD in civilians? This small study looks at the symptom frequencies in established cases of PTSD in 31 police officers and 72 civilians and compares the two. No significant differences were found, although there was a tendency for higher use of alcohol and to try not to think about the trauma. Police officers had more experiences of reliving the trauma or acting as if the trauma were recurring but, again, not to a significant degree. PMID- 14741079 TI - Efficacy of rofecoxib and nimesulide in controlling postextraction pain in oral surgery: a randomised comparative study. AB - Rofecoxib 50 mg/day for 6 days provided better postoperative analgesia than nimesulide 200 mg/day in a randomised trial in patients (n = 130) undergoing surgical extraction of third molars. The superiority of rofecoxib over nimesulide was especially marked during the first 2-3 postoperative days and in patients with fully impacted molars. The drugs had similar effects on clinical signs of local postoperative inflammation. The effectiveness of rofecoxib in this study, plus considerations of the toxicity profile of nimesulide, support the conclusion that rofecoxib is preferable to nimesulide for relief of post-operative pain in patients undergoing surgical extraction of molars. PMID- 14741080 TI - The value of post-marketing medication surveys in Parkinson's disease. PMID- 14741081 TI - Levodopa therapy with entacapone in daily clinical practice: results of a post marketing surveillance study. AB - The catechol-O-methyl transferase inhibitor entacapone is given in combination with levodopa/dopa decarboxylase inhibitor for Parkinson's disease (PD) patients experiencing end-of-dose wearing-off. This 4-week post-marketing surveillance study was undertaken to assess patients' responses to levodopa combined with entacapone in a real clinical practice setting. Overall, 466 patients with idiopathic PD treated with levodopa and experiencing symptoms of wearing-off were recruited. Both physicians and patients recorded the response to therapy, including improvements and side-effects. Following initiation of entacapone treatment, the average daily levodopa dose was reduced from 510 to 453 mg. Physician assessment of entacapone efficacy was judged to be "very good" or "good" in 77.6% of the patients, and tolerability was considered to be "very good" or "good" in 92.4% of patients, with only 12 patients (2.6%) withdrawing from the study. Compared with baseline, there was a decrease in the mean duration of daily 'off' time from 3.0 to 1.3 h per day during the treatment period. Adverse events were in line with those previously reported, with diarrhoea being the most frequent event. The percentage of patients suffering from dyskinesia decreased from 46 to 34%, and of those patients still suffering from dyskinesia, the average daily duration of dyskinesia was reduced from 2.2 to 1.7 h. The use of adjunct dopamine agonists decreased from 67 to 59%. At study end, the percentage of patients who rated their quality of life (QoL) as "very good" or "good" increased from 12.1 to 51.7% and the percentage of patients who rated their QoL as "bad" or "very bad" decreased from 40 to 10.7%. In summary, the results of this survey conducted in real clinical practice support the findings of previous clinical trials demonstrating the efficacy and tolerability of entacapone, as well as the benefits of improved QoL, for patients achieved with entacapone. PMID- 14741082 TI - Impact of tranexamic acid vs. aprotinin on blood loss and transfusion requirements after cardiopulmonary bypass: a prospective, randomised, double blind trial. AB - INTRODUCTION: Aprotinin (AP) reduces blood loss and transfusions after cardiopulmonary bypass (CPB), but may sensitise patients and is expensive. Tranexamic acid (TA) has less side-effects, but data regarding its efficacy are controversial. The aim of our prospective, randomised, double-blind study was to compare the impact of AP vs. TA on drainage blood loss and transfusion requirements in patients undergoing first time CABG on CPB. MATERIALS AND METHODS: One hundred and twenty adult patients were randomised to receive either high-dose AP according to Hammersmith or a total of 2 g TA. Perioperative blood products were transfused in a standardised fashion. Blood loss was measured up to 24 h. Demographic and clinical patient data were collected until hospital discharge. RESULTS: The data from 118 patients (TA: n = 58, AP: n = 60) who completed the study according to protocol were analysed. Blood loss at 24 h postoperation in TA patients was significantly higher (896 +/- 354 ml) as compared to AP patients (756 +/- 347 ml; p = 0.03). TA patients received 1.5 +/- 1.5 units of red blood cells (AP: 1.5 +/- 1.7, p = 1.0), 1.3 +/- 2.0 units of fresh frozen plasma (AP: 1.0 +/- 2.0, p = 0.38) and 0.5 +/- 1.4 units of platelets (AP: 0.2 +/- 0.7, p = 0.15). Clinical data, including perioperative myocardial infarction rate, acute renal failure, mechanical ventilation, hospital stay and mortality, were not significantly different between either group. CONCLUSION: Our data show a difference in blood loss between TA and high-dose AP. Although statistically significant, it has little clinical relevance, because perioperative transfusion requirements were similar for both groups. Thus, TA appears to be a cost-effective alternative to AP in primary CABG patients. PMID- 14741083 TI - [Necessary elements for the modern exercise of medical sciences]. PMID- 14741084 TI - [Hypophyseal macroadenomas. A neurosurgical challenge]. AB - Management of pituitary adenoma is a real challenge because of neurovascular structures that surround the sellar region. In this paper we included 336 patients with pituitary tumors greater than 10 mm in diameter to make an analysis of surgical results. In this group, a discrete female predominance was found, with an average of 43.2 years of age and 21.5 mm in diameter of lesions. The clinical course was dominated by headache and visual deficit and only one quarter of patients had functioning tumors. The majority of the tumors were operated on by a transsphenoidal approach; in 72% of cases, the lesion was completely removed. Visual recovery was observed in 68% of patients, better in those who had a shorter clinical course; on the other hand, only 34 of 84 patients with functioning tumors could be cured. Complications were specially related to growing pattern of adenoma and its consistency. Even though higher morbidity and mortality was found in patients operated on by craniotomy, they also had the most complex lesions. Based on our experience, we are proposing here some recommendations to chose correctly the best surgical option and to avoid complications. Finally, we designed a diagram showing the critical route for optimal treatment of these lesions. PMID- 14741085 TI - [Clinical-radiologic correlation in chronic sinusitis]. AB - INTRODUCTION: Chronic sinusitis is a pathology that has been increasing in incidence in world population. The Mexican Institute of Social Security (IMSS) reports 91 cases per year per 100,000 people. In the last decades, much has been advanced concerning knowledge of the physiology of paranasal sinuses and thus of the pathogenesis of sinusitis; this has led to developing of functional endoscopic sinus surgery. To perform this kind of surgery, it is mandatory to have computerized tomography, and the necessity of clinical-radiologic correlation has been recognized to attain better diagnosis and treatment. OBJECTIVE: To determine clinical-radiologic correlation of chronic sinusitis in adult patients. MATERIALS AND METHODS: Correlational, transverse, convenience assigned blind study from January to July 2002 in the Department of Otolaryngology, Hospital de Especialidades at the Centro Medico Nacional Siglo XXI in Mexico City. Kendall test was applied to a group of 31 patients. RESULTS: Nasal obstruction was found in 87%, mucopurulent rhinorrea in 55%, mucosal edema in 74%, and polyps in 42%. Most frequently paranasal sinus affected by sinusitis was maxillary sinus in 97% and least affected was frontal, in 58%. Of tomographic findings, most frequent was obstruction of ostiomeatal complex in 84%. There was no statistical significance (p > 0.05) in clinical-radiologic. DISCUSSION: No correlation was found between signs and symptoms evaluated with tomographic findings, as reported by other authors. PMID- 14741086 TI - [Primary hyperparathyroidism: a report of 67 cases. The experience of 5 years at the IMSS Centro Medico Nacional Siglo XXI Specialties Hospital]. AB - INTRODUCTION: Primary hyperparathyroidism (PHPT) is a frequent disease. It is characterized by hypercalcemia and elevation of parathyroid hormone (PTH) levels. The incidence is variable, being more frequent in women (3:1) in the perimenopauseal period. In the last decades, diagnosis of asymptomatic PHPT has increased due to automatized calcium determinations. Duration of the disease and serum concentrations of calcium and PTH determine symptomatology. Surgery is the definitive treatment. It is important to assess surgical criteria in asymptomatic patients. Successful treatment of this disease depends on a highly specialized team of parathyroid surgeons. MATERIAL AND METHODS: We reviewed the clinical and biochemical background of patients subjected to parathyroidectomy from January 1998 to August 2003 in our medical center. We then analyzed clinical and laboratory data, final histopathologic diagnosis, and surgery outcome. RESULTS: Sixty seven patients were included, with an average age of 50 years (+/- 11.9), 15 men and 52 women. Greatest frequency of PHPT was observed in patients between 40 and 60 years of age. Average calcium value was 11.7 mg/dl (+/- 1.23) and that of PTH was 194.5 pg/ml (+/- 93). Histopathologic diagnosis was adenoma in 56 (83.58%), hyperplasia in 9 (13.43%), and carcinoma in 2 (2.9%) patients. Cure of the disease was achieved in 65 patients (97%). PHPT is a surgically curable disease; it is necessary to confirm diagnosis and decide on surgical treatment in accordance with established criteria even if the disease is asymptomatic. PMID- 14741087 TI - [Diagnostic evaluation of fiberopticbronchoscopy in bronchogenic carcinoma in neoplasic-suspicious cases]. AB - Fiberopticbronchoscopy (FOB) is a well established method for diagnosis in bronchogenic cancer (CABR). OBJECTIVE: To evaluate sensitivity and specificity of FOB in suspicious cases of lung neoplasia. MATERIAL AND METHODS: A total of 119 suspicious cases of CABR were subjected to FOB under local anesthesia with lidocaine and sedation and bronchial brushing and lavage were carried out. In all evident lesions, a biopsy was taken. Sputum was collected in carbowax for 5 days. When histopathogic and cytopathologic studies were negative, fine needle aspiration (FNA) of the lesion was performed; if pleural involvement was present, a biopsy and a sample of liquid were taken. RESULTS: Of 119 cases, diagnosis of CABR was confirmed by FOB in 45: brushing 25; lavage 25, and biopsy 29 with sensitivity 52.33%, specificity 100%, PPV 100%, and NPV, 42.31%. In 17, positivity was obtained in two-thirds samples; in carbowax, three were positive. In another 41 CABR-suspicious cases, all FOB samples were negative and neoplasia was confirmed by FNA in 15, biopsy of extrathoracic lymphatic node in 11, pleural biopsy in eight and liquid cytopathology in six. Non-malignant pathology was found in 33 cases: pneumonia 22, lung abscess three and tuberculosis, eight. FBO is indicated in CABR-suspicious cases. Negativity of FOB samples is due to different causes. All FOB instruments must be in optimal operative conditions and must be handled by a well-trained endoscopist. PMID- 14741088 TI - [Pulmonary mechanics, oxygenation index, and alveolar ventilation in patients with two controlled ventilatory modes. A comparative crossover study]. AB - OBJECTIVE: To compare in a crossover study pulmonary mechanics, oxigenation index (PaO(2)/FiO(2)), and partial pressure of CO(2) in arterial blood (PaCO(2)) in patients with mechanical ventilation in two controlled ventilatory modes. SETTING: Intensive care unit of a university affiliated hospital. DESIGN: Prospective crossover clinical trial. PATIENTS AND METHODS: A total 114 consecutive patients were admitted to the intensive care unit (ICU) under controlled mechanical ventilation with SaO(2) >90% and FiO(2) <0.5 and assigned by random allocation to either volume control (VC) and constant inspiratory flow (square flow curve) (group I) or pressure control mode (PC) (group II). Both groups were ventilated with tidal volume (Vt) of 7 ml/kg, respiratory rate (RR) 14/min, inspiratory-expiratory ratio 1:2 (I:E), PEEP 5 cm H(2)O, and FiO(2) 0.4. After 15 min of mechanical ventilation, pulmonary mechanics, oxygenation index (OI), and PaCO(2) were measured and registered, and ventilatory mode was switched to PC mode in group I and to VC in group II, maintaining the same ventilator settings. Pulmonary mechanics, OI, and PaCO(2) were again registered after 15 min of ventilation. RESULTS: Peak inspiratory pressure (PIP) was higher in VC than in PC (31.5 vs 26 cm H(2)O), which resulted in a significant increase in transpulmonary pressure amplitude difference (DP) (25 vs 19 cm H(2)O). Mean airway pressure (MAP) and OI were lower in VC than in PC (11.5 vs 12 cm H(2)O, and 198.5 vs 215, respectively). Dynamic compliance (DynC) was lower in VC than in PC (20 vs 26 ml/cm H(2)O), p < 0.05 for all values. At constant ventilator settings in the same patient, PC and not VC ventilation decreases PIP (which results in smaller transpulmonary pressure amplitude difference), increases MAP, and DynC and improves the oxygenation index. PMID- 14741089 TI - [Preliminary report. Usefulness of computed tomographic angiography in the protocol of a kidney donor]. AB - OBJECTIVE: To determine clinical correlation of reports of computed tomographic angiography renal (CT-AR) and surgical findings of the kidney donor patient. MATERIAL AND METHODS: Patients were submitted nephrectomy in the related live donor renal transplant program between January and December 2002 as paut of life to which he is made as he CT-AR study protocol. Statistical analysis was carried out by descriptive statistics. RESULTS: Anatomical characteristics of 35 kidneys of the same number of live donors (AD) submitted CT-AR were evaluated and comparison with report of surgical technique was made. Incidence of accessory renal arteries was 23%. As reported by CT-AR, the were 39 renal arteries (91%) compared with 43 arteries found during surgery. CT-AR identified four supernumerary renal arteries (50%) of eight identified during surgical technique; 36 hiliar arteries (90%) and three polar arteries were identified by CT-AR (100%). Only one a case report of early bifurcation of renal artery (20%) by CT AR was recorded. Anatomical characteristics of veins were described in their totality. CT-AR is a useful instrument to identify alterations in anatomical structure of the renal vasculature, with results similar to other studies for description of renal arteries and veins. We propose ATR as the initial study for evaluation of the renal architecture of the live kidney (LKD). PMID- 14741090 TI - [Open-groin hernia repair utilizing E-PTF mesh technique]. AB - BACKGROUND: At present, utilization of mesh is best option for surgical groin hernia treatment. Affirmation must be corroborated in hernia clinics of a national concentration hospital to ascertain which is the most recommendable with regard to technique therapeutics and economic factors. PLACE: A tertiary level concentration general hospital. MATERIAL: There were 61 groin hernia repairs in 59 patients, 54 males 91.5% and five females (8.5%). Average 43 age was, minimum 17, maximum 81, median 41 mode 55 years. Plain mesh of expanded polytetrafluoroethylene was used. METHODS: A prospective, descriptive, and longitudinal study was conducted in a 1-year. Inclusion criteria included patients who accepted the mesh proceeding and bought the mesh. We made a date gathering sheet with age, gender, hospital stay, surgical time, pain scale and complications. Patients underwent follow-up at 7, 14, 21, and 30 days after each month. RESULTS: Average surgical time was 53 min average hospital stay 52 h. Five patients had complications, urine retention was present in two, and serosity in one there was case of rejection of surgical absorbable suture not related with mesh. There was no prolonged hospital stay, and no relapses. Pain measurement was performed at two different times; Average pain index was 3.76 for first and 2.48 for second on a 1-10 scale. On average, there were two doses of analgesics per patient. In this group, surgery was well accepted without severe complications and with low scale of pain. PMID- 14741091 TI - [Post total laryngectomy stomal recurrence. Case report and review of the literature]. AB - Development of stomal recurrence following total laryngectomy is a devastating scenario with an extremely poor prognosis. Overall rate of stomal recurrence ranges from 1.7%-40%, with an average rate of 7.5%. Irrespective of the etiology of stomal recurrence, it invariably consists of diffuse infiltration of tumor into the soft tissues of the neck and mediastinum, thereby making control of the disease difficult. Despite aggresive surgery, radiotherapy, and chemotherapy, salvage rate is poor. Prevention of stomal recurrence is therefore of paramount importance and the only means of reducing incidence. Systematic use of preventive surgical measures together with postoperative radiotherapy to the stoma and superior mediastinum have led to decrease in appearance of stomal recurrence. Intensive follow-up should be performed for patients with laryngeal carcinoma who had subglottic involvement, paratracheal lymph node metastasis, or both to detect stomal recurrence at an early stage. PMID- 14741092 TI - [Critical points for success in supracricoid partial laryngectomy with cricohyoidepiglottoepexy]. AB - Cancer of the larynx is a relatively rare neoplasia in Mexico as well as in the rest of the world; however, evolution of treatment in recent years showed important results in terms of survival and local control. Supracricoid partial laryngectomy with cricohyoidoepiglottopexy is a surgical technique that permits similar oncologic results to other conservatives techniques for glottic carcinoma, with excellent functional results. The objective of the present work was to show the critical points of the technique based on the description of Laccourreye, emphasizing the anatomical aspect of resection and reconstruction to avoid complications and achieve best results. PMID- 14741093 TI - [Iatrogenesis. Individual error? System failure?]. AB - Iatrogenesis has been considered for a long time as a situation created by the wrongdoing of an individual, that is, the physician. Nevertheless, we propose that medical error be considered a very complex "social omission" in which public funding, training of health professionals including administrators, and even the public education may be responsible for an adverse medical event, considering that medical officials are just a part of a system. Detection of latent errors and the epidemiologic study of those that have occurred, including differences in quality, must be considered as the main effort in prevention. PMID- 14741094 TI - [Official commentary on the work entitled: "Iatrogenesis. Individual error? System failure?"]. PMID- 14741095 TI - [Investigation and ethics in pediatrics. General considerations]. AB - Considerations are put forth with regard to legal aspects of investigation in human beings and the importance of informed consent as established in the general Law of Health. Here in are pointed out legal and ethical dispositions established in the General Law of Health for protection of minors and the disabled. Commentary is presented on importance of information, autonomy, understanding, and freedom of election that are not afforded to those under the legal age and there interfere in minors being included in investigation projects. Diverse ethical aspects are commented on that should be considered throughord the entire investigation. PMID- 14741096 TI - Doublecortin and a tale of two serines. AB - Doublecortin (DCX) is a microtubule-associated protein that interacts with and regulates the microtubule cytoskeleton and is required for neuronal migration in the cortex. Two papers in this issue of Neuron (Schaar et al. and Tanaka et al.) demonstrate a role for phosphorylation in the regulation of Doublecortin. Together with recent results showing that Doublecortin may play a role regulating the morphology of migrating neurons, these findings provide new insight into the mechanisms governing neuronal migration. PMID- 14741097 TI - Context matters. AB - Midbrain dopamine neurons are thought to encode the difference between predicted and actual reward on conditioning tasks. Successful models assumed a simple form of prediction that depended only on currently available information. In this issue of Neuron, Nakahara and colleagues record from dopamine neurons in alert monkeys and show that the neurons can encode predictions that are not so restricted, taking into account the context of past trends. PMID- 14741098 TI - Language abilities of motor cortex. AB - A new exploration of the cortical network underlying our language abilities by Hauk et al., in this issue of Neuron, shows that the process of giving meaning to words differentially activates the motor cortex according to the semantic category of the word. PMID- 14741099 TI - Frisking the whiskers: patterned sensory input in the rat vibrissa system. AB - How are two prominent environmental features, surface texture and object location, transduced and encoded as rats whisk? Recent papers show that textures may excite intrinsic mechanical vibrations of the vibrissae. Although these vibrations are too rapid to be directly followed by cortical neurons, there is evidence that their speed is encoded by contact-dependent sensory signals. In addition to contact, sensory signals exist that report the angular position of the vibrissae. The combination of contact and reference signals may be used to decode spatial variations in the environment, particularly the location of objects in head-centered coordinates. PMID- 14741100 TI - A molecular mechanism for stabilization of learning-induced synaptic modifications. AB - Olfaction is a principal sensory modality in rodents, and rats quickly learn to discriminate between odors and to associate odor with reward. Here we show that such olfactory discrimination (OD) learning consists of two phases with distinct cellular mechanisms: an initial NMDAR-sensitive phase in which the animals acquire a successful behavioral strategy (rule learning), followed by an NMDAR insensitive phase in which the animals learn to distinguish between individual odors (pair learning). Rule learning regulates the composition of synaptic NMDARs in the piriform cortex, resulting in receptors with a higher complement of the NR2a subunit protein relative to NR2b. Rule learning also reduces long-term potentiation (LTP) induced by high-frequency stimulation of the intracortical axons in slices of piriform cortex. As NR2a-containing NMDARs mediate shorter excitatory postsynaptic currents than those containing NR2b, we suggest that learning-induced regulation of NMDAR composition constrains subsequent synaptic plasticity, thereby maintaining the memory encoded by experience. PMID- 14741101 TI - ApoE and clusterin cooperatively suppress Abeta levels and deposition: evidence that ApoE regulates extracellular Abeta metabolism in vivo. AB - Apolipoprotein E (apoE) and clusterin can influence structure, toxicity, and accumulation of the amyloid-beta (Abeta) peptide in brain. Both molecules may also be involved in Abeta metabolism prior to its deposition. To assess this possibility, we compared PDAPP transgenic mice that develop age-dependent Abeta accumulation in the absence of apoE or clusterin as well as in the absence of both proteins. apoE(-/-) and clusterin(-/-) mice accumulated similar Abeta levels but much less fibrillar Abeta. In contrast, apoE(-/-)/clusterin(-/-) mice had both earlier onset and markedly increased Abeta and amyloid deposition. Both apoE(-/-) and apoE(-/-)/clusterin(-/-) mice had elevated CSF and brain interstitial fluid Abeta, as well as significant differences in the elimination half-life of interstitial fluid Abeta measured by in vivo microdialysis. These findings demonstrate additive effects of apoE and clusterin on influencing Abeta deposition and that apoE plays an important role in regulating extracellular CNS Abeta metabolism independent of Abeta synthesis. PMID- 14741102 TI - Doublecortin microtubule affinity is regulated by a balance of kinase and phosphatase activity at the leading edge of migrating neurons. AB - Doublecortin (Dcx) is a microtubule-associated protein that is mutated in X linked lissencephaly (X-LIS), a neuronal migration disorder associated with epilepsy and mental retardation. Although Dcx can bind ubiquitously to microtubules in nonneuronal cells, Dcx is highly enriched in the leading processes of migrating neurons and the growth cone region of differentiating neurons. We present evidence that Dcx/microtubule interactions are negatively controlled by Protein Kinase A (PKA) and the MARK/PAR-1 family of protein kinases. In addition to a consensus MARK site, we identified a serine within a novel sequence that is crucial for the PKA- and MARK-dependent regulation of Dcx's microtubule binding activity in vitro. This serine is mutated in two families affected by X-LIS. Immunostaining neurons with an antibody that recognizes phosphorylated substrates of MARK supports the conclusion that Dcx localization and function are regulated at the leading edge of migrating cells by a balance of kinase and phosphatase activity. PMID- 14741103 TI - Cdk5 phosphorylation of doublecortin ser297 regulates its effect on neuronal migration. AB - Mutations in the doublecortin (DCX) gene in human or targeted disruption of the cdk5 gene in mouse lead to similar cortical lamination defects in the developing brain. Here we show that Dcx is phosphorylated by Cdk5. Dcx phosphorylation is developmentally regulated and corresponds to the timing of expression of p35, the major activating subunit for Cdk5. Mass spectrometry and Western blot analysis indicate phosphorylation at Dcx residue Ser297. Phosphorylation of Dcx lowers its affinity to microtubules in vitro, reduces its effect on polymerization, and displaces it from microtubules in cultured neurons. Mutation of Ser297 blocks the effect of Dcx on migration in a fashion similar to pharmacological inhibition of Cdk5 activity. These results suggest that Dcx phosphorylation by Cdk5 regulates its actions on migration through an effect on microtubules. PMID- 14741104 TI - A CaMKII-NeuroD signaling pathway specifies dendritic morphogenesis. AB - The elaboration of dendrites is fundamental to the establishment of neuronal polarity and connectivity, but the mechanisms that underlie dendritic morphogenesis are poorly understood. We found that the genetic knockdown of the transcription factor NeuroD in primary granule neurons including in organotypic cerebellar slices profoundly impaired the generation and maintenance of dendrites while sparing the development of axons. We also found that NeuroD mediated neuronal activity-dependent dendritogenesis. The activity-induced protein kinase CaMKII catalyzed the phosphorylation of NeuroD at distinct sites, including endogenous NeuroD at Ser336 in primary neurons, and thereby stimulated dendritic growth. These findings uncover an essential function for NeuroD in granule neuron dendritic morphogenesis. Our study also defines the CaMKII-NeuroD signaling pathway as a novel mechanism underlying activity-regulated dendritic growth that may play important roles in the developing and mature brain. PMID- 14741105 TI - Recordings from single neocortical nerve terminals reveal a nonselective cation channel activated by decreases in extracellular calcium. AB - Synaptic activity causes reductions in cleft [Ca(2+)] that may impact subsequent synaptic efficacy. Using modified patch-clamp techniques to record from single neocortical nerve terminals, we report that physiologically relevant reductions of extracellular [Ca(2+)] ([Ca(2+)](o)) activate voltage-dependent outward currents. These outward currents are carried by a novel nonselective cation (NSC) channel that is indirectly inhibited by various extracellular agents (rank order potency, Gd(3+) > spermidine > Ca(2+) > Mg(2+), typical for [Ca(2+)](o) receptors). The identification of a Ca(2+) sensor-NSC channel pathway establishes the existence of a mechanism by which presynaptic terminals can detect and respond to reductions in cleft [Ca(2+)]. Activation of NSC channels by falls in [Ca(2+)](o) would be expected during periods of high activity in the neocortex and may modulate the excitability of the presynaptic terminal. PMID- 14741106 TI - Bidirectional modification of presynaptic neuronal excitability accompanying spike timing-dependent synaptic plasticity. AB - Correlated pre- and postsynaptic activity that induces long-term potentiation is known to induce a persistent enhancement of the intrinsic excitability of the presynaptic neuron. Here we report that, associated with the induction of long term depression in hippocampal cultures and in somatosensory cortical slices, there is also a persistent reduction in the excitability of the presynaptic neuron. This reduction requires postsynaptic Ca(2+) elevation and presynaptic PKA and PKC-dependent modification of slow-inactivating K(+) channels. The bidirectional changes in neuronal excitability and synaptic efficacy exhibit identical requirements for the temporal order of pre- and postsynaptic activation but reflect two distinct aspects of activity-induced modification of neural circuits. PMID- 14741107 TI - Dopamine neurons can represent context-dependent prediction error. AB - Midbrain dopamine (DA) neurons are thought to encode reward prediction error. Reward prediction can be improved if any relevant context is taken into account. We found that monkey DA neurons can encode a context-dependent prediction error. In the first noncontextual task, a light stimulus was randomly followed by reward, with a fixed equal probability. The response of DA neurons was positively correlated with the number of preceding unrewarded trials and could be simulated by a conventional temporal difference (TD) model. In the second contextual task, a reward-indicating light stimulus was presented with the probability that, while fixed overall, was incremented as a function of the number of preceding unrewarded trials. The DA neuronal response then was negatively correlated with this number. This history effect corresponded to the prediction error based on the conditional probability of reward and could be simulated only by implementing the relevant context into the TD model. PMID- 14741108 TI - Modulation of caudate activity by action contingency. AB - Research has increasingly implicated the striatum in the processing of reward related information in both animals and humans. However, it is unclear whether human striatal activation is driven solely by the hedonic properties of rewards or whether such activation is reliant on other factors, such as anticipation of upcoming reward or performance of an action to earn a reward. We used event related functional magnetic resonance imaging to investigate hemodynamic responses to monetary rewards and punishments in three experiments that made use of an oddball paradigm. We presented reward and punishment displays randomly in time, following an anticipatory cue, or following a button press response. Robust and differential activation of the caudate nucleus occurred only when a perception of contingency existed between the button press response and the outcome. This finding suggests that the caudate is involved in reinforcement of action potentially leading to reward, rather than in processing reward per se. PMID- 14741109 TI - Perisaccadic mislocalization orthogonal to saccade direction. AB - Saccadic eye movements transiently distort perceptual space. Visual objects flashed shortly before or during a saccade are mislocalized along the saccade direction, resembling a compression of space around the saccade target. These mislocalizations reflect transient errors of processes that construct spatial stability across eye movements. They may arise from errors of reference signals associated with saccade direction and amplitude or from visual or visuomotor remapping processes focused on the saccade target's position. The second case would predict apparent position shifts toward the target also in directions orthogonal to the saccade. We report that such orthogonal mislocalization indeed occurs. Surprisingly, however, the orthogonal mislocalization is restricted to only part of the visual field. This part comprises distant positions in saccade direction but does not depend on the target's position. Our findings can be explained by a combination of directional and positional reference signals that varies in time course across the visual field. PMID- 14741110 TI - Somatotopic representation of action words in human motor and premotor cortex. AB - Since the early days of research into language and the brain, word meaning was assumed to be processed in specific brain regions, which most modern neuroscientists localize to the left temporal lobe. Here we use event-related fMRI to show that action words referring to face, arm, or leg actions (e.g., to lick, pick, or kick), when presented in a passive reading task, differentially activated areas along the motor strip that either were directly adjacent to or overlapped with areas activated by actual movement of the tongue, fingers, or feet. These results demonstrate that the referential meaning of action words has a correlate in the somatotopic activation of motor and premotor cortex. This rules out a unified "meaning center" in the human brain and supports a dynamic view according to which words are processed by distributed neuronal assemblies with cortical topographies that reflect word semantics. PMID- 14741111 TI - Biosensing and drug delivery at the microscale. PMID- 14741112 TI - Closed-loop insulin delivery-the path to physiological glucose control. AB - The development of an artificial pancreas for the treatment of insulin-dependent diabetes is a highly desired endeavor for patients, physicians, scientists, and engineers. Historical algorithms and recent progress in research and technology are reviewed in the present article, together with aspects of beta-cell physiology that lead to normal glucose tolerance. Algorithms are evaluated for their ability to deliver insulin as to recreate, as closely as possible, glucose and insulin profiles observed in healthy individuals. Emphasis is placed upon how the algorithms compare to the beta-cell's secretory response, specifically first phase and second-phase insulin secretion. Experimental closed-loop data employing intravenous and subcutaneous glucose sensors and implanted and external insulin pumps (Medtronic MiniMed, Northridge CA) are presented. PMID- 14741113 TI - Hard and soft micromachining for BioMEMS: review of techniques and examples of applications in microfluidics and drug delivery. AB - Recent development in microfabrication (micromachining, microelectromechanical systems, MEMS) permits the integration of hard and soft structures, and enables the design of controllable microfluidic systems, which may be applied to drug delivery. In this paper, we present a tutorial review of both classical "hard" and more recent "soft" micromachining techniques. We then provide examples where these techniques are combined to produce hydrogel-based microfluidic control systems. The most complex of these systems utilizes a very small hydrogel based on phenylboronic acid to control the flow of an insulin solution in response to changes in glucose concentration. PMID- 14741114 TI - Electronic MEMS for triggered delivery. AB - Implantable electronic devices such as pacemakers and neural implants are often used for electrical stimulation. The usage of microfabrication techniques to produce microelectromechanical systems (MEMS) has allowed engineers to address a wider range of clinical indications. A new direction in the area of MEMS technology is the goal of achieving pulsatile drug delivery. The digital capabilities of MEMS may allow greater temporal control over drug release compared to traditional polymer-based systems, while the batch-processing techniques used in the microelectronics industry can lead to greater device uniformity and reproducibility than is currently available to the pharmaceutical industry. A repertoire of structures, including microreservoirs, micropumps, valves, and sensors, is being developed that will provide a strong foundation for the design of integrated, responsive MEMS for drug delivery. PMID- 14741115 TI - Integrated microsystems for controlled drug delivery. AB - Efficient drug delivery and administration are needed to realize the full potential of molecular therapeutics. Integrated microsystems that incorporate extremely fast sensory and actuation capabilities can fulfill this need for efficient drug delivery tools. Photolithographic technologies borrowed from the semiconductor industry enable mass production of such microsystems. Rapid prototyping allows for the quick development of customized devices that would accommodate for diverse therapeutic requirements. This paper reviews the capabilities of existing microfabrication and their applications in controlled drug delivery microsystems. The next generation of drug delivery systems--fully integrated and self-regulating--would not only improve drug administration, but also revolutionize the health-care industry. PMID- 14741116 TI - Flow control with hydrogels. AB - With the advent of the genomic revolution and the sequencing of the human genome complete, the majority of pharmaceuticals under development are proteins. Consequently, new techniques to more effectively administer these new protein therapeutics need to be developed. One method that is gaining popularity in the research community involves the use of responsive hydrogel actuators for flow control in drug delivery devices. Responsive hydrogels are materials able to undergo a volume change in response to a stimulus from their local environment. The following paper overviews recent advances made using hydrogel actuators for flow control such as resistance based valves, hydrogel jacket valves, hybrid hydrogel membrane valve, electrically triggered valves, and biomimetic valves. Also reviewed are several hydrogel flow control systems such as a flow sorter and pH-regulation system. The chemistry of the hydrogel actuators can be tweaked to allow physiological variables to trigger the volume expansion of the hydrogel actuators as demonstrated by several glucose sensitive hydrogel valves reviewed below. Therefore, the door to physiological feedback controlling the infusion rate in a drug delivery device is opened and has the potential to revolutionize protein pharmaceutical drug delivery. PMID- 14741117 TI - Micromachined biocapsules for cell-based sensing and delivery. AB - In recent years, rapid advancements have been made in the biomedical applications of micro- and nanotechnology. While the focus of such technology has primarily been on in vitro analytical and diagnostic tools, more recently, in vivo therapeutic and sensing applications have gained attention. The long-term integration of cells with inorganic materials provides the basis for novel delivery and sensing platforms. Our recent work has focused on the ability to maintain cells long term in nanoporous silicon-based microenvironments. This paper describes the creation of monodisperse, nanoporous, biocompatible, silicon membranes as a platform for the delivery of cells. Studies described herein focus on the interaction of silicon-based substrates with cells of interest in terms of viability, proliferation, and functionality. Such microfabricated nanoporous membranes can be used both in vitro for cell-based assays and in vivo for immunoisolation and drug delivery applications. PMID- 14741118 TI - Proceedings of the 1st ssDNA Viruses of Plants, Birds, Pigs and Primates meeting. Saint-Malo, France, 24-27 September 2001. PMID- 14741119 TI - Molecular biology of Porcine circovirus: analyses of gene expression and viral replication. AB - The rep gene of Porcine circovirus type 1 directs the synthesis of two proteins. The full-length protein Rep is 312 amino acids in size, the spliced variant Rep' is truncated (168 aa) and exon 2 is frame-shifted. Replication of PCV1 DNA depends on synthesis of both proteins. Rep and Rep' bind in vitro to double stranded DNA fragments comprising part of the origin of replication of PCV1, but the minimal binding sites of the two proteins are distinct. Rep protein represses the promoter of the rep gene by binding to the two inner hexamers H1 and H2. Although Rep' binds to the same sequence, it does not influence Prep. Twelve hours after PCV1 infection, similar amounts of rep and rep' were detected by real time PCR, but later on, the ratio of the two transcripts varied. Both proteins are co-localised in the nucleus and formation of homo- and heteromeric complexes has been observed. When a replication assay was performed, in which Rep and Rep' protein of PCV1 was used to replicate the origin of PCV1 and PCV2, the rep gene products were found to initiate replication at both origins of replication. PMID- 14741120 TI - Chicken anemia virus induced apoptosis: underlying molecular mechanisms. AB - In 1990, the chicken anemia virus (CAV) genome was cloned by us and proven to be representative for CAV isolates worldwide. This genome contains unique promoter/enhancer replication elements and genes. Upon infection of its target cells, CAV replicates via a double-stranded (ds) DNA intermediate. From this ds CAV molecule, a single mRNA is transcribed, which encodes for three distinct proteins VP1, VP2, and VP3 or apoptin. Its capsid contains only the VP1 protein. However, for the production of the neutralizing epitope, co-synthesis of VP1 and VP2 are needed. CAV genomes with mutations in the 12 bp insert of the promoter/enhancer region were shown to produce immunogenic functional CAV particles. Mutations in these and other regulatory elements of CAV might also decrease its virus load resulting in a reduced pathogenic effect. CAV causes fatal cytopathogenic effects in e.g. chicken thymocytes via apoptosis. Under in vitro conditions, CAV replicates only in transformed chicken cell lines, which indicates that at least a part of the CAV life-cycle requires transformed-like cellular events. In these transformed cell lines, the synthesis of the apoptin protein alone mimics the CAV-induced apoptosis, whereas the VP2 protein also harbors some apoptotic activity. Extensive studies on apoptin resulted in the characterization of domains essential for its apoptotic activity and nuclear localization, which seems to be related with its ability to induce apoptosis. Therefore, both VP2 and apoptin are of interest in reducing the pathogenicity of CAV infections. A series of biomedical studies on apoptin have been carried out in human cell systems, which are informative about the mechanism of CAV-induced apoptosis in chicken (transformed) cells. Synthesis of apoptin alone induces apoptosis in various human transformed and/or tumorigenic cell lines, but not in normal human diploid cells. A striking difference in the cellular localization of apoptin was observed in human normal diploid cells versus tumor cells. In all tumor cells, apoptin is located mainly in the heterochromatic regions of the nucleus, whereas in normal cells it is present in peri-nuclear structures. Apoptin contains a bipartite nuclear localization signal, and one domain that resemble a nuclear export signal. Elucidation of parts of the apoptin-induced apoptotic pathway revealed unique characteristics: apoptin-induced apoptosis is independent of the tumor suppressor p53. The anti-apoptotic protein Bcl-2 does not inhibit but even accelerates apoptin-induced apoptosis in tumor cells, whereas over expression of Bcl-2 in normal cells has no effect on the apoptin activity. Upstream caspases are not involved, whereas downstream caspase 3 is, but seems not to be essential. A number of novel proteins were shown to interact with apoptin in transformed cells. Future studies of apoptin, VP2 and related cellular proteins in chicken cells will unravel the regulatory aspects of CAV induced apoptosis. PMID- 14741121 TI - Human circoviruses. AB - TT virus (TTV) and TTV-like mini virus (TLMV) represent the first described human circoviruses. They do not show significant sequence homology with any other animal circovirus identified to date. They are both detected with high prevalences in various body fluids. The spread mode may include the parenteral way, the transmission by saliva droplets and the fecal-oral route. Genetic variability within a viral group is high and the co-infection by distinct viral strains is common in a given individual. The description of several messenger RNAs obtained after multiple splicing revealed a specific transcription profile. Despite apparent asymptomatic infections, the possible association of variants of TTV and TLMV with a given pathology cannot be formally ruled out. PMID- 14741122 TI - Nanoviruses: genome organisation and protein function. AB - Nanoviruses, single-stranded DNA (ssDNA) plant viruses with a multipartite genome, share similarities with members of the Circoviridae family that infect mammals or birds as well as with the Geminiviridae, the only other plant virus family with circular ssDNA genomes. Although the virions of the latter are unique and different from that of the circoviruses, the mode of replication of viruses with monopartite or multipartite circular ssDNA genomes is strikingly similar. They multiply by rolling circle replication using virus-encoded multifunctional replication initiator proteins (Rep proteins) that catalyse initiation of ssDNA replication and resolution of replicative ssDNA into circular single-stranded virion DNA. All these ssDNA viruses exploit host polymerases for DNA synthesis and code for proteins that modulate the host's cell cycle favourably for virus multiplication. Recent three-dimensional structure analyses of a geminivirus and a parvovirus Rep protein have revealed an intriguing similarity between the catalytic domains of their respective Rep proteins. Furthermore, these structural data revealed that ssDNA virus replication initiator proteins might represent evolutionary intermediates between certain RNA-binding proteins and some multifunctional origin-binding proteins of papovaviruses. PMID- 14741123 TI - Geminivirus DNA replication and cell cycle interactions. AB - The Geminiviridae family includes a large number of viruses that infect plants and have a unique geminate virion particle, a single-stranded genome of approximately 2.6-3.0 kb, and replicate through a rolling-circle mechanism. Since they encode for just a few proteins (4-6 depending on the members that belong to four different genera), a rich variety of interactions has evolved between viral proteins and host factors to develop the virus replicative cycle. Among them, we have been particularly interested so far: (i). in the interference with cell cycle regulatory proteins of the retinoblastoma-related (RBR)/E2F pathway and (ii). in the interaction with host DNA replication factors necessary for the assembly of a functional replication complex at the viral origin of DNA replication during the rolling-circle stage. Yeast two-hybrid assays revealed that wheat dwarf virus RepA protein, but nor Rep protein, interacts with plant RBR protein. Interestingly, deletion of the C-terminal domain of Rep confers the truncated protein the ability to interact with RBR, suggesting that this domain may hinder the LXCXE RBR-binding motif. Secondary structure predictions support such a possibility. PMID- 14741124 TI - Subviral DNAs associated with geminivirus disease complexes. AB - Ageratum conyzoides (ageratum) is a common and widespread weed species that may act as a reservoir host for geminivirus diseases. Ageratum plants growing in Singapore and exhibiting yellow vein disease are infected with a complex mixture of viral, subviral and recombinant DNA components. This whitefly-transmitted disease is caused by the monopartite begomovirus ageratum yellow vein virus (AYVV) in association with a recently discovered satellite component referred to as DNA beta. Diseased plants also contain a subviral component, referred to as DNA 1, that has probably become associated with the begomovirus and adapted to whitefly transmission during mixed infection with an aphid-transmitted nanovirus. Unlike DNA beta, the nanovirus-like component is not essential for the disease. Recombination between the viral and subviral DNAs occurs frequently and may play an important evolutionary role by generating component diversity. The identification of a similar complex associated with cotton leaf curl disease (CLCuD), a serious constraint to cotton growing in Pakistan, and the isolation of DNA beta homologues from diverse plant species growing in widespread geographical locations suggests that such disease complexes are common and may have a significant impact on agriculture in the eastern hemisphere. PMID- 14741125 TI - The clinical expression and emergence of porcine circovirus 2. AB - Since its discovery and characterization in western Canada in 1995, the distribution of post-weaning multisystemic wasting syndrome (PMWS) has grown and the syndrome is undoubtedly recognized worldwide. More recently, there is a heightened interest in several potentially related conditions including porcine dermatitis and nephropathy syndrome (PDNS), proliferative and necrotizing pneumonia (PNP), congenital tremors (CT-AII), perinatal myocarditis and reproductive failure. Although the causality of porcine circovirus type 2 (PCV2) has recently been proven [Vet. Pathol. 37 (2000) 254; J. Comp. Pathol. 122 (2000) 9; J. Vet. Diagn. Invest. 13 (2001) 185], PCV2 is present in both diseased and healthy pig populations. Furthermore, recent serologic studies have shown that there are no significant differences in PCV2 specific IgG titres between PMWS clinical and non-clinical herds (Harding et al., in press). Clearly, the transmission, epidemiology, and factors triggering clinical expression are poorly understood. With these uncertainties in mind, the objective of this paper is to review the clinical expression and emergence of PMWS and the potentially associated disease syndromes including PDNS. PMID- 14741126 TI - Pathological findings associated with naturally acquired porcine circovirus type 2 associated disease. AB - Porcine circovirus type 2 (PCV2) is a novel virus of the Circoviridae family which is considered the cause of postweaning multisystemic wasting syndrome (PMWS). PCV2 has also been associated to a number of pathological conditions of pigs, including porcine dermatitis and nephropathy syndrome, reproductive failure, porcine respiratory disease complex, proliferative and necrotising pneumonia and congenital tremor type AII. Pathological studies have been used to describe and characterise PMWS and these emerging conditions associated with PCV2. The objective of this review is to concentrate on the gross, microscopic and ultrastructural pathology associated with natural cases of PCV2 associated disease, along with some speculations on the pathogenesis of naturally occurring PMWS. PMID- 14741127 TI - Immunosuppression in postweaning multisystemic wasting syndrome affected pigs. AB - The present review concentrates on the clinical, pathological and immunological aspects of pigs suffering from PMWS which strongly suggest that PCV2 may be, in particular conditions, a cause of secondary immunodeficiency in pigs. From a clinical point of view, the lack of antibiotic therapy response against the disease, the existence of a litter effect and the concurrence of other disease syndromes and well-known secondary pathogens, such as Pneumocystis carinii, Chlamydia spp. and Aspergillus spp., may account as features of immunosuppression in PMWS. Furthermore, pathologic, immunohistologic and flow cytometric studies also suggest that pigs with PMWS may be immunosuppressed. Lymphocyte depletion of follicular and interfollicular areas together with macrophage infiltration of lymphoid tissues is a unique lesion, which is the basic feature of PMWS affected pigs. These findings are highly correlated with the decrease of circulating B- and T-cells and the diminution of these cell types in lymphoid organs, and with the increase of macrophage/monocytes lineage cells both in peripheral blood and lymphoid tissues in both naturally and experimentally PMWS affected pigs. The altered populations of cells participating in the immune system response both in blood and tissues suggests, at least in those severely PMWS affected pigs, a transient inability of diseased pigs to mount an effective immune response. From these points of view, strong suspicions on the immunosuppressive status of PMWS affected pigs do exist; however, future studies are needed to characterise the exact role of PCV2 on the immune system of pigs affected with PMWS. PMID- 14741128 TI - Porcine circovirus-2 and concurrent infections in the field. AB - Porcine circovirus-2 (PCV-2) is the necessary cause of post-weaning multisystemic wasting syndrome (PMWS) in swine; however, a variety of co-factors, including other infectious agents, are thought to be necessary in the full expression of disease. Porcine parvovirus (PPV) was found in the inoculum used in the first experiments to reproduce PMWS in gnotobiotic swine. Retrospective and prospective studies in the field and laboratory have demonstrated PCV-2 can act synergistically with PPV to enhance the severity of PMWS. PCV-2 has been shown to play a role in the porcine infectious disease complex (PRDC). Other co-infecting agents with PCV-2 in the lung include, porcine reproductive and respiratory syndrome virus (PRRSV), swine influenza virus (SIV) and Mycoplasma hyopneumoniae. Exposure of pregnant sows to PPV, PRRSV, or encephalomyocarditis virus may interact with PCV-2 infected foetuses. The severity of hepatic lesions in PCV-2 infected pigs may be enhanced by co-infection with agents such as swine hepatitis E virus and Aujezsky's disease virus. Additional studies are required to determine the mechanistic basis for the interaction of PCV-2 with other agents in the pathogenesis of the various clinical syndromes that have been associated with PCV-2 infection. PMID- 14741129 TI - PMWS: experimental model and co-infections. AB - Porcine circovirus type 2 (PCV2) is now recognised as the causal agent of porcine multisystemic wasting syndrome (PMWS), an economically important wasting disease of young pigs [J. Vet. Diagn. Invest. 12 (2000) 3]. Gross lesions of PMWS include generalised lymphadenopathy, hepatitis, nephritis and pneumonia and typical histological lesions include lymphocytic depletion and multinucleated giant cell formation in lymph nodes, degeneration and necrosis of hepatocytes, and multifocal lymphohistocytic interstitial pneumonia. This communication will review the results of experimental infections of gnotobiotic (GN), colostrum deprived (CD) and colostrum-fed (CF) pigs within our group, and elsewhere, with PCV2 and the conclusions that can be drawn from this work. PMID- 14741130 TI - Avian circovirus diseases: lessons for the study of PMWS. AB - The diseases associated with psittacine beak and feather disease virus (BFDV), pigeon circovirus (PiCV) and goose circovirus (GoCV), which can be classified with porcine circovirus type 2 (PCV2) as members of the genus Circovirus of the family Circoviridae, have clinico-pathological features in common with post weaning multisystemic wasting syndrome (PMWS), with which PCV2 infection is causally associated. Intracytoplasmic botryoid inclusions within macrophages and depletion of T and B lymphocytes are common histopathological features, and, in each case, affected animals usually exhibit ill-thrift and a predisposition to secondary infections, that is suggestive of an underlying immunosuppression. Although these avian diseases have been the subjects of relatively little research, their study can provide directly applicable lessons in the areas of diagnosis, epidemiology, pathogenesis and disease control for those charged with investigating PMWS. In keeping with its taxonomic separation as the only member of the genus Gyrovirus, the disease caused by chicken anaemia virus (CAV) differs histopathologically from the other circovirus-associated diseases. Most notably, the target cells of CAV have been identified as haemocytoblasts and precursor T lymphocytes, with lymphocyte depletion, which affects T cells only, occurring in cells directly infected with the virus. Nonetheless, CAV is the best-researched circovirus and provides excellent examples of both virus-induced immunosuppression and virus-virus interactions. The study of CAV-induced disease can therefore provide valuable, if less directly applicable lessons. PMID- 14741131 TI - Viremia and effect of fetal infection with porcine viruses with special reference to porcine circovirus 2 infection. AB - This publication reviews some pathogenetic features of the transplacental infection with porcine viruses in sows. Viremia either with virus freely circulating or associated to peripheral blood mononuclear cells (PBMC) is an essential part of such pathogenesis. Virus replication occurs either in fetal tissues only or both in fetal and maternal tissues and the outcome may be different. Since porcine circovirus 2 (PCV2) has been associated with reproductive failure in sows, the question was asked what type of viremia PCV2 causes and what the effect of PCV2 is on the pregnant uterus. Seronegative gilts were oronasally inoculated and plasma and PBMC were monitored for infectious virus and for quantity of viral DNA copies. Infectious virus was found in plasma only at 21 days post-inoculation (DPI). Virus associated to PBMC was detected between 14 and 49 DPI. Viral DNA was found in plasma between 14 and 49 DPI and associated to PBMC between 7 and 63 DPI (end of experiment). Direct intra-fetal inoculation at 57, 75 and 92 days of gestation and collection of fetuses 21 days later showed that the virus replicates highly in fetal tissues, particularly in the heart. Fetal death occurred in the 57 days sows while virus and antibodies were observed in the 75- and 92-day inoculated sows. Inoculation at 57 and 75 days of gestation and collection of the piglets at the end of pregnancy showed that intrauterine spread had occurred to fetuses adjacent to the inoculated ones and that fetal death occurred also in the presence of antibodies. The pregnancy was not interrupted.This study shows that PCV2 causes viremia which is largely cell-associated and that virus replication in fetuses causes fetal death with mummification. Whether such transplacental infection occurs in the immune sow population is questionable. PMID- 14741132 TI - Extensive mast cell degranulation in bovine respiratory syncytial virus associated paroxystic respiratory distress syndrome. AB - Bovine respiratory syncytial virus (BRSV) infection is an important cause of outbreaks of respiratory disease among calves. This virus commonly induces mild to severe respiratory signs but, in a substantial proportion of cases, is also reported to be associated with paroxystic respiratory distress syndrome (PRDS). The pathogenesis of this 'malignant' clinical form has not been fully elucidated. The present study aimed at determining whether mast cell (MC) degranulation plays a role in the physiopathologic cascade leading to the PRDS. Paired serum samples were taken in herds during outbreaks of severe respiratory diseases (acute sera) and 3 weeks after (convalescent sera). Based on seroconversion to BRSV and clinical picture, 67 pairs of sera were selected from calves with a BRSV associated PRDS for circulating MC tryptase determination. A MC metachromatic score was measured in post-mortem lungs from animals died from a BRSV-associated PRDS (principals) and compared with reference scores obtained from healthy lungs (controls). Levels of tryptase were significantly higher in acute sera (26.6 +/- 12.4 microg/l) compared to convalescent sera (8.4 +/- 7.8 microg/l; P<0.001). Metachromatic scores yielded significantly different results between controls and principals (P<0.01), demonstrating a significant disappearance of metachromatic granules from lung MCs in principals. Taken together, these data demonstrate the presence of an extensive MC degranulation in BRSV-associated PRDS. PMID- 14741133 TI - Expression of immune response genes in rainbow trout skin induced by Gyrodactylus derjavini infections. AB - By means of semi-quantitative RT-PCR, expression of a number of immune relevant genes was studied in skin of small rainbow trout Oncorhynchus mykiss (Walbaum, 1792) fry during both primary and secondary infections with the ectoparasitic monogenean Gyrodactylus derjavini Mikailov, 1975. The target genes studied included the cyto- and chemokines TNF-alpha1, TNF-alpha2, TGF-beta and IL-8, the inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX-2) genes and finally, two cell markers, the beta-chains of TCR and MHC II, from the adaptive arm of the immune system. In general, constitutive expression of all studied genes was apparent. Significant increases in expression of the TNF-alpha1 isoform could be observed at day 8 p.i. in primary infections and although less marked, the alpha2 isoform of TNF showed a similar trend. With the cytokine TGF-beta, 8 10 times increase in the transcription levels was observed in secondary infections compared to uninfected hosts. However, no parasite related changes in expression patterns could be observed for IL-8. Parasite infections elicited strong iNOS expression by 4 days p.i., but significant differences were not detected before day 8 p.i., when transcript levels were increased 5.5-9.6 times compared to uninfected controls. Augmented expression of COX-2 could also be observed in primary, but not secondary, infections at later stages of infections. No clear parasite related changes in transcript levels of the two cell markers TCRbeta and MHC IIbeta could be observed. Although the cellular source(s) was not determined, most of the examined factors appear to take part in a local signalling network of pivotal importance for the initiation, orchestration, effectuation and modulation of immune responses in rainbow trout against the ectoparasite G. derjavini. PMID- 14741134 TI - Immunoperoxidase studies of cell mediated immune effector cell populations in early Mycobacterium avium subsp. paratuberculosis infection in sheep. AB - Immunoperoxidase (IPX) labelling for CD4, CD8, TCR-gammadelta, WC1, CD1b, IFN gamma, CD45R, CD56 and lysozyme was used to investigate changes in cell mediated immune effector cell populations in the intestinal Peyer's patches (PP) and mesenteric lymph nodes of lambs, 2 and 4 months after experimental infection with low doses of sheep strain Mycobacterium avium subsp. paratuberculosis (M. a. paratuberculosis). The organism was cultured from the tissues of each infected lamb, but histological lesions were not present. This infection model was considered to be more representative of natural M. a. paratuberculosis infection than previous studies. Infected sheep had significantly more CD4+ cells in the mucosa, domes and interfollicular areas of the terminal ileum, and in the interfollicular areas of the jejunal Peyer's patch. Infected sheep also had significantly increased numbers of TCR-gammadelta+ cells in the mucosa and interfollicular areas of the jejunal Peyer's patch, and increased numbers of WC1+ cells in the ileal Peyer's patch. These findings are consistent with previous findings in sheep given higher doses of cattle strain M. a. paratuberculosis. Significantly fewer CD1b+ cells were present in the paracortical areas of the mesenteric lymph nodes of infected sheep, and the reduction was greater in sheep infected for 4 months compared to sheep infected for only 2 months. Down regulation of CD1b expression may be important for the continued survival and multiplication of M. a. paratuberculosis as specific adaptive immunity develops. Across all sheep, jejunal Peyer's patches had higher numbers of CD4+, CD8+, TCR gammadelta+, WC1+ and CD45R+ cells, and lower numbers of CD56+ fibres compared to ileal Peyer's patches. These findings confirm and extend the peculiarities of the terminal ileal Peyer's patch in the young ruminant, with possible implications for the early establishment of M. a. paratuberculosis infection. PMID- 14741135 TI - The sequential analysis of local inflammatory cells during abomasal nematode infection in periparturient sheep. AB - A technique to take sequential tissue biopsy samples in multiparous, periparturient ewes from the abomasal mucosa is described, developed in parallel in Scotland and New Zealand. Samples were extracted via abomasal cannulae inserted into the wall of the abomasum and exteriorised through dorso-ventral laparotomy. Animals recovered quickly post-surgery, and tolerated the cannula and sampling without any adverse signs of pain or discomfort. The technique was deployed in two pilot studies to investigate the sequential mucosal inflammatory cell responses in well-defined parasitological models, during the periparturient relaxation of immunity in ewes infected with gastrointestinal nematodes and subjected to different feeding treatments. One experiment (Moredun Research Institute, Scotland) involved the infection of twin-bearing ewes with Teladorsagia circumcincta L3 either before, or after lambing. By feeding ewes with different levels of protein supplementation, preliminary data on the impact of nutrition on the eosinophil, mucosal mast cell and globule leucocyte responses during this period were investigated. A similar study was also performed at Lincoln University, New Zealand, to investigate these cell responses in sheep fed relatively high or low protein diets during pregnancy, and infected with a combined immunisation regime of T. circumcincta and Trichostrongylus colubriformis L3. These studies confirmed the phenomenon termed the periparturient relaxation in immunity (PPRI) where a transitory increase in faecal egg counts is observed during late pregnancy and lactation, and this effect was exacerbated during protein undernutrition. Although the number of animals was low in each experiment and the cell responses variable, the results together suggest a reduction in the number of mucosal mast cells and globule leucocyte during the PPRI when protein supply was restricted. The present paper thus describes a successful technique to monitor ovine mucosal cell populations during local immune responses in normal and pregnant sheep. It is envisaged that this technique will be a powerful adjunct to investigations into mucosal immune mechanisms and disease pathogenesis, and will be employed to confirm the influence of dietary protein on the local inflammatory cell responses during the PPRI. PMID- 14741136 TI - Cetacean-reconstituted severe combined immunodeficient (SCID) mice respond to vaccination with canine distemper vaccine. AB - Morbillivirus infections have been responsible for mass mortalities in several species of marine mammals. Nevertheless, relatively little is known on the pathogenesis of the disease and the immune response to the agent, especially in cetaceans, hindering the treatment of individuals and the development of appropriate vaccines, given the difficulty of performing experimental work in marine mammals. The reconstitution of severe combined immunodeficient (SCID) mice, which do not have the ability to reject grafts, with lymphocytes from different species has been used with increasing success as a surrogate species model to study the immune system. We injected NOD/SCID mice with lymphocytes from different species of cetaceans and further vaccinated those mice with a commercial canine distemper virus (CDV) vaccine to develop a practical model to study cetacean immune response to a morbillivirus. Reconstitution was detected in 10/20 mice reconstituted with harbor porpoise spleen, 6/10 mice reconstituted with harbor porpoise lymph node cells, 8/10 mice reconstituted with fresh beluga PBMCs and none of the mice reconstituted with neonate bottlenose dolphin spleen or thymus cells when assessed 42-63 days after reconstitution. While a humoral immune response was detected in none of the reconstituted mice, a cell-mediated immune response to the CDV vaccine was detected in 6/15 (40%) and 2/18 (11%) of the SCID mice after reconstitution with cetacean immune cells after a single or booster vaccination, respectively, for a combined total of 8/33 (24%). This represents the first demonstration of successful reconstitution of SCID mice with marine mammal cells, and to the authors' knowledge, the first direct demonstration of a primary antigen-specific cell-mediated immune response in reconstituted SCID mice. This model will be useful for further research on the physiology of the marine mammal immune system and its response to infectious agents and vaccines, with possible important outcomes in conservation issues. PMID- 14741137 TI - Genomic characterization of equine interleukin-4 receptor alpha-chain (IL4R). AB - Three overlapping fragments of the equine interleukin-4 receptor alpha chain gene (IL4R) were cloned and sequenced. The resulting 3553 bp cDNA sequence exhibited homology to human, murine and bovine IL4R. The equine IL4R exhibits many conserved features when compared to other species, including intron-exon boundary positions and amino acid sequence motifs characteristic of type I cytokine receptors. The IL4R gene was localized to horse chromosome ECA13 by synteny mapping on a somatic cell hybrid panel. Evidence for an alternative splice variant of IL4R was found in the genomic sequence and subsequently verified using RT-PCR on equine monocyte RNA. A polymorphism screen of the largest exon, homologous to exon 12 of the human IL4R gene, was performed using DNA from 60 horses of various breeds which yielded 11 coding-region single nucleotide polymorphisms (SNPs), 7 synonymous and 4 non-synonymous. Three of the four non synonymous SNPs occur at high frequencies and are found very near a conserved tyrosine residue. PMID- 14741138 TI - Characterization of F21.A, a monoclonal antibody that recognize a leucocyte surface antigen for killer whale homologue to beta-2 integrin. AB - The specificity of F21.A, a monoclonal antibody raised against bottlenose dolphin leucocytes, was characterized in killer whale on the basis of immunoprecipitation of a protein of 94 kDa, as well as flow cytometric analysis. While minimally expressed on resting cells, F21.A labeled a homologue to beta-2 integrin in 89 97% of PMA-activated neutrophils, 53-66% of activated monocytes, and activated B cells but not T cells. Activation of neutrophils reached its maximum 10 min after PMA stimulation. F21.A did not label intracellular stores as did both cross reacting anti-canine CD11b and CD18, suggesting that an activation-induced conformational change would expose a neoepitope recognized by F21.A. F21.A labeling was largely inhibited by pre-incubation with plasma, suggesting a binding site closely related to that for fibrinogen. In vitro phagocytosis and respiratory burst were almost fully inhibited upon pre-incubation with F21.A, demonstrating its functional importance. This antibody is foreseen as a possible valuable diagnostic and research tool in cetacean immunology. PMID- 14741139 TI - Reactive oxygen and nitrogen intermediates contribute to Haemophilus somnus lipooligosaccharide-mediated apoptosis of bovine endothelial cells. AB - Although Haemophilus somnus causes septicemia and vasculitis in cattle, relatively little is known about how H. somnus affects endothelial cells in vitro. We previously reported that H. somnus lipooligosaccharide (LOS)-induced activation of caspases-3, -8 and -9, and apoptosis of bovine pulmonary artery endothelial cells (BPAEC) in vitro. Previous reports indicate that the generation of reactive oxygen species (ROS) or reactive nitrogen intermediates (RNI) can contribute to the induction of apoptosis. In the present study, we sought to determine whether ROS and RNI are involved in LOS-mediated apoptosis of BPAEC. We found that H. somnus LOS induced the generation of ROS in BPAEC, which was blocked by pretreatment with membrane permeable ROS scavengers, such as dimethylsulfoxide (DMSO) and allopurinol (AP). Addition of DMSO or AP significantly reduced H. somnus LOS-mediated caspase-3 activation. Addition of membrane impermeable ROS scavengers (e.g. catalase and superoxide dismutase), failed to block LOS-mediated caspase-3 activation, suggesting a role for intracellular generation of ROS in LOS-induced apoptosis of BPAEC. Addition of N(G)-nitro-L-arginine methyl ester (L-NAME) or aminoguanidine, which are selective inhibitors of nitric oxide synthase, blocked NO release and significantly reduced caspase-3 activation in LOS treated BPAEC. These data suggest H. somnus LOS triggers endogenous ROS and RNI production by endothelial cells, which contributes to apoptosis. PMID- 14741140 TI - Production of a functional chicken single-chain variable fragment antibody derived from caecal tonsils B lymphocytes against macrogamonts of Eimeria tenella. AB - Avian coccidiosis is due to a protozoan intracellular parasite belonging to the genus Eimeria which multiplies in the intestine of the host. In order to identify Eimeria antigens which reflect the natural avian humoral immune response, chicken hybridomas were produced by fusion of myeloma MuH1 with B lymphocytes from Eimeria tenella infected chicken. B lymphocytes used for fusions were isolated from tonsils at the basis of caeca where the parasite develops. One of the clones (G1F5) recognised oocyst antigens and the macrogamont stage of the parasite in ELISA and immunofluorescence assay. A single-chain variable fragment (scFv) antibody was cloned from the light chain variant region (VL) and heavy chain variant region (VH) genes of the hybridoma. This recombinant antibody (scFv G1F5) exhibited antigen binding specificity to oocysts and macrogamonts of E. tenella equivalent to the mAb produced by the clone G1F5. Nucleotide sequence analysis of VL genes from scFv G1F5 compared to the germ-line revealed vestiges of gene conversion. scFv derived from chicken B lymphocytes isolated from the gut associated lymphoid tissue following experimental infection can reveal specific antigens recognised by the avian immune response. PMID- 14741141 TI - Immunohistochemical characterization of inflammatory cell populations and adhesion molecule expression in synovial membranes from dogs with spontaneous cranial cruciate ligament rupture. AB - This study describes the distribution of CD4+ and CD8alpha+ T lymphocytes, B lymphocytes, macrophages, MHC class II antigens, immunoglobulin (IgG, IgM, IgA) containing cells and of adhesion molecules belonging to the CD11/CD18 family in synovial membrane biopsies from 28 dogs with spontaneous rupture of the cranial cruciate ligament (CCL). Synovial membranes from 11 dogs without evidence of joint lesions were used as control tissues. The main cell types in synovial membranes from dogs with CCL rupture were B lymphocytes and plasma cells belonging to the IgG isotype. The severity of inflammatory cell infiltration in CCL cases was positively correlated with the expression of adhesion molecules. Double immunofluorescence labelling of frozen sections revealed that in the inflamed synovium of dogs with CCL rupture numerous dendritic cells expressing MHC class II antigen and canine CD1c were present. The findings further support the view that in the synovium of dogs with CCL rupture an immunologic response is going on in which dendritic cells are possibly involved by presenting hitherto unknown antigens to T lymphocytes. PMID- 14741142 TI - Standardized case definitions of adverse events following immunization (AEFI). PMID- 14741143 TI - Fever as an adverse event following immunization: case definition and guidelines of data collection, analysis, and presentation. PMID- 14741144 TI - Generalized convulsive seizure as an adverse event following immunization: case definition and guidelines for data collection, analysis, and presentation. PMID- 14741145 TI - Hypotonic-Hyporesponsive Episode (HHE) as an adverse event following immunization: case definition and guidelines for data collection, analysis, and presentation. PMID- 14741146 TI - Acute intussusception in infants and children as an adverse event following immunization: case definition and guidelines of data collection, analysis, and presentation. PMID- 14741147 TI - Nodule at injection site as an adverse event following immunization: case definition and guidelines for data collection, analysis, and presentation. PMID- 14741148 TI - Persistent crying in infants and children as an adverse event following immunization: case definition and guidelines for data collection, analysis, and presentation. PMID- 14741150 TI - Avipox-based simian immunodeficiency virus (SIV) vaccines elicit a high frequency of SIV-specific CD4+ and CD8+ T-cell responses in vaccinia-experienced SIVmac251 infected macaques. AB - The ability of ALVAC- or fowlpox-based simian immunodeficiency virus (SIV) vaccines to boost SIV-specific CD4+ and CD8+ T-cell responses was tested in 10 vaccinia-experienced macaques infected with SIVmac251. The CD8+ T-cell response to the dominant Gag(181-189) CM9 was quantitated in seven Mamu-A*01-positive macaques by tetramer staining, by ex vivo cytotoxic T-lymphocyte (CTL) activity, and by intracellular cytokine staining (ICS) with the specific Gag(181-189) CM9 peptide. The overall CD8+ T-cell response to Gag was assessed using a peptide pool encompassing the entire Gag protein followed by measurement of TNF-alpha production in ICS assay. Similarly, virus-specific CD4+ T-cell responses were measured by ICS for TNF-alpha following stimulation with the Gag-overlapping peptide and by proliferative response following stimulation with purified p27 Gag. The two vaccine modalities effectively boosted both CD4+ and CD8+ SIV specific T-cell response despite prior exposure to the vaccinia-derivative NYVAC vector, suggesting that sequential boosting with either avipox-based vector vaccine candidate is a realistic approach in immune therapy of human immunodeficiency virus type 1 (HIV-1)-infected individuals. PMID- 14741151 TI - Long-term persistence of anti-HBs after vaccination against HBV: an 18 year experience in health care workers. AB - The aim of the present study was to evaluate the long-term persistence of seroprotection after hepatitis B virus (HBV) vaccination. A total of 422 health care workers (HCWs) were evaluated 4.8-18.8 years after primary immunization (mean follow-up 11.8 years); 241 of them had received plasma-derived vaccines and 181 had been given yeast-derived vaccines; 107 subjects received a booster dose of yeast-derived vaccine 6 years after primary immunization with either plasma derived or yeast-derived vaccines. Seroprotection was assumed when the anti-HBs titers were >10 mIU/ml. The overall response after primary immunization was 98.8%. Among subjects who reached a 10 year follow-up, those treated with plasma derived vaccine had a seroprotection rate of 87.8 compared to 81.6% of those vaccinated with yeast-derived vaccines (P<0.001). Anti-HBs geometric mean titers (GMTs) after primary immunization were similar in the two groups, but were significantly lower at 10 years follow-up in the group that had received a yeast derived vaccine (104 mIU/ml versus 244 mIU/ml in those who used a plasma-derived vaccine, P<0.05). Anti-HBs GMTs in the 107 subjects given the booster dose were 242 mIU/ml pre-booster titer, and rose to 35,171 mIU/ml after the booster dose. A mean 10.1 years after the booster dose, GMTs were 952 mIU/ml. Overall, the anti HBs seroprotection rate was 95.4% (102 subjects). Based on GMT results, no booster dose is necessary in healthy adults for at least 10 years after primary immunization. PMID- 14741152 TI - Prevention of intestinal amebiasis by vaccination with the Entamoeba histolytica Gal/GalNac lectin. AB - Prevention of intestinal infection by Entamoeba histolytica would block both invasive disease and parasite transmission. The amebic Gal/GalNAc lectin mediates parasite adherence to the colonic surface and fecal anti-lectin IgA is associated with protection from intestinal reinfection in children. We tested if vaccination with the E. histolytica Gal/GalNAc lectin could prevent cecal infection in a C3H mouse model of amebic colitis. Two trials using native lectin purified from the parasite and two trials using a 64 kDa recombinant fragment ("LecA") were performed with a combined intranasal and intraperitoneal immunization regimen using cholera toxin and Freund's adjuvants, respectively. Two weeks after immunization mice were challenged intracecally with trophozoites, and 4-12 weeks after challenge mice were sacrificed for histopathologic evaluation of infection. Vaccination prevented intestinal infection with efficacies of 84 and 100% in the two native lectin trials and 91 and 34% in the two LecA trials. Mice with detectable pre-challenge fecal anti-lectin IgA responses were significantly more resistant to infection than mice without fecal anti-lectin IgA responses. These results show for the first time that immunization with the Gal/GalNAc lectin can prevent intestinal amebiasis in mice and suggest a protective role for fecal anti lectin IgA in vivo. PMID- 14741153 TI - Vaccination against the nematode Haemonchus contortus with a thiol-binding fraction from the excretory/secretory products (ES). AB - Fractionated excretory/secretory products (ES) of adult Haemonchus contortus were evaluated as protective antigens. The proteins were successively eluted from a Thiol Sepharose column using 25 mM cysteine followed by 25 mM Dl-dithiothreitol (DTT). Sheep were vaccinated three times and challenged with 5000 third stage infective larvae (L3) of H. contortus. Highest level of protection was found in sheep vaccinated with the DTT-eluted fraction in which egg output and worm burden were reduced by 52 and 50%, respectively, compared to the adjuvant control group. There was a positive correlation between fecundity (number of eggs per female) and the cumulative EPG or worm burden. Serum and mucus antibody levels of ES specific immunoglobulins increased after immunizations and after challenge for IgG, IgA and IgE. The harvesting of H. contortus from animals clustered per group revealed the presence of cysteine protease activity in the ES of all groups but in addition to that, metalloprotease activity was also detected in the groups vaccinated with the DTT-eluted fraction, total ES and adjuvant only, in contrast to previous batches of ES (completely inhibited by E64) obtained from non vaccinated animals. PMID- 14741154 TI - Stability of mono- and trivalent meningococcal outer membrane vesicle vaccines. AB - The stability during storage of outer membrane vesicles (OMVs) of Neisseria meningitidis group B was studied. Three types of OMVs were compared for their stability, containing either one (monovalent) or three different PorA subtypes (trivalent), the latter with and without class 4 outer membrane protein (OMO, RmpM). Aqueous formulations were stored freeze-dried (4 degrees C), frozen (-70 degrees C) and in liquid form at 4, 37 and 56 degrees C. Physico-chemical properties and immunogenicity of the OMVs as well as PorA conformation and antigenicity (P1.7-2,4, the subtype present in all formulations) were monitored during 1 year. At -70 or 4 degrees C, the structure and immunogenicity of OMVs was preserved. Storage of OMVs at high temperatures (37 or 56 degrees C) induced destruction of the OMV structure and denaturation of PorA, followed by chemical degradation. Immunogenicity decreased or was lost completely. Changes observed in the fluorescence spectra of degraded OMVs were also seen in tryptophan (Trp) and tyrosine (Tyr) derivatives incubated at 56 degrees C, indicating the occurrence of chemical degradation of tryptophan and tyrosine residues in PorA. Trivalent OMVs were slightly more stable at 37 degrees C than monovalent OMVs as assessed by in vitro methods, but these differences did not result in differences in the immunogenicity. The stability of trivalent OMVs was not affected by the presence of RmpM. Both trivalent and monovalent OMVs could be freeze-dried with preservation of their immunogenicity. In conclusion, OMVs are sensitive to elevated temperatures, but are stable in the frozen or freeze-dried state or when stored at 4 degrees C in the liquid state. PMID- 14741155 TI - Maternally derived humoral immunity to bovine viral diarrhea virus (BVDV) 1a, BVDV1b, BVDV2, bovine herpesvirus-1, parainfluenza-3 virus bovine respiratory syncytial virus, Mannheimia haemolytica and Pasteurella multocida in beef calves, antibody decline by half-life studies and effect on response to vaccination. AB - The passive immunity transferred to calves from their dams was investigated in a beef herd to determine half-life of antibody, estimated time to seronegative status and effect on immunization. One hundred two beef calves in a commercial ranch under standard management conditions were utilized. Samples were collected at branding (day 0). This was the first possible date to collect samples postcalving. This was approximately 2 months postcalving, and days 95 and 116. The calves were divided into two groups: vaccinates (51) and nonvaccinates (51). The calves were vaccinated with a commercial inactivated viral vaccine containing bovine viral diarrhea virus (BVDV)1a, BVDV2, bovine herpesvirus-1 (BHV-1), parainfluenza-3 virus (PI-3V), and bovine respiratory syncytial virus (BRSV) on days 0 and 95. Half of the vaccinated and unvaccinated calves also received one dose of an experimental Mannheimia haemolytica and Pasteurella multocida vaccine at day 95. Serums were tested for neutralizing antibody titers to BVDV1a, BVDV1b, BVDV2, BHV-1, PI-3V, and BRSV. Antibodies were detected by ELISA to M. haemolytica whole cell, M. haemolytica leukotoxin, and P. multocida outer membrane protein (OMP). The mean half-life of viral antibodies in nonvaccinated calves to each virus was: BVDV1a, 23.1 days (d); BVDV1b, 22.8 d; BVDV2, 22.9 d; BHV-1, 21.2 d; PI-3V, 30.3 d; and BRSV, 35.9 d. The mean half-life of viral antibodies was greater for vaccinates than for nonvaccinates for all viruses except BRSV. The calculated mean time to seronegative status for nonvaccinates based on titers at day 0 was: BVDV1a, 192.2 d; BVDV1b, 179.1 d; BVDV2, 157.8 d; BHV-1, 122.9 d; PI-3V, 190.6 d; and BRSV, 186.7 d. There was an active immune response after vaccination with two doses to all the viruses, except BRSV. Mean antibody titers of vaccinates at day 116 were statistically higher than nonvaccinates for all viruses except BRSV. However on an individual calf basis there were few seroconversions (four-fold rise or greater to BVDV1a, BVDV1b, BVDV2, PI-3V, or BRSV; or two-fold rise for BHV-1) in the presence of viral antibodies. The predicted time of seronegative status for a group of calves for vaccination programs may not be appropriate as there may be a range of titers for all calves at day 0. In this study the range for BVDV1a was 16-16,384; BVDV1b, 8 8192; BVDV2, 0-8192; BHV-1, 0-935; PI-3V, 8-2048; and BRSV, 8-4096. Using the half-life of 23 d for BVDV1a, the time thereafter for seronegative status would be 46 and 299 d compared to the calculated date of 192.2 d using the mean of estimated time to seronegative status for all the calves. There was an active humoral response in the vaccinated calves to M. haemolytica and P. multocida. Cowherd humoral immunity based on serum antibodies should be monitored as it may relate to transfer of maternal antibodies to calves. Exceptionally high levels of viral antibodies transferred to calves could interfere with the antibody response to vaccination. PMID- 14741156 TI - Early DNA vaccination of puppies against canine distemper in the presence of maternally derived immunity. AB - Canine distemper (CD) is a disease in carnivores caused by CD virus (CDV), a member of the morbillivirus genus. It still is a threat to the carnivore and ferret population. The currently used modified attenuated live vaccines have several drawbacks of which lack of appropriate protection from severe infection is the most outstanding one. In addition, puppies up to the age of 6-8 weeks cannot be immunized efficiently due to the presence of maternal antibodies. In this study, a DNA prime modified live vaccine boost strategy was investigated in puppies in order to determine if vaccinated neonatal dogs induce a neutralizing immune response which is supposed to protect animals from a CDV challenge. Furthermore, a single DNA vaccination of puppies, 14 days after birth and in the presence of high titers of CDV neutralizing maternal antibodies, induced a clear and significant priming effect observed as early as 3 days after the subsequent booster with a conventional CDV vaccine. It was shown that the priming effect develops faster and to higher titers in puppies preimmunized with DNA 14 days after birth than in those vaccinated 28 days after birth. Our results demonstrate that despite the presence of maternal antibodies puppies can be vaccinated using the CDV DNA vaccine, and that this vaccination has a clear priming effect leading to a solid immune response after a booster with a conventional CDV vaccine. PMID- 14741157 TI - Incorporation of CpG oligodeoxynucleotide fails to enhance the protective efficacy of a subunit vaccine against Mycobacterium tuberculosis. AB - Vaccines which offer better protection than BCG are now badly needed for controlling tuberculosis infection throughout the world. Immunological adjuvants capable of inducing a TH1 type of protective response are necessary to augment the immune response, particularly in the case of subunit vaccines. It is now well established that oligodeoxynucleotides (ODN) containing cytidine phosphate guanosine (CpG) motifs enhance cell-mediated responses in vivo by increasing the production of the TH1 cytokines IL-12 and interferon gamma (IFNgamma). To determine if this would improve subunit vaccination of mice CpG ODN were added to a subunit vaccine consisting of the culture filtrate proteins (CFP) of Mycobacterium tuberculosis H37Rv. It was observed that although adding CpG ODN to the vaccines promoted substantially increased IFNgamma production by lymph node cells draining sites of inoculation, this failed to translate after aerosol challenge into any degree of enhancement of bacterial clearance in the lungs, influx of IFN-positive T cells, or changes in histopathology. These data suggest that the vaccine enhancing effects of CpG ODN are relatively transient. PMID- 14741158 TI - DNA immunization of mice with a plasmid encoding Neisseria gonorrhea PorB protein by intramuscular injection and epidermal particle bombardment. AB - Immunogenicity of a DNA vaccine encoding PorB from Neisseria gonorrhoeae strain FA1090 was analyzed in BALB/C mice immunized by intramuscular needle injection or epidermal gene gun bombardment. Both delivery routes generated measurable specific antibodies although the gene gun response was slower. Antibody isotypes were indicative of Th2 activation following gene gun immunization and of Th1 activation following intramuscular injection. In both immunization protocols, boosting with either renatured recombinant (rr) PorB protein or PorB expressed from Venezuelan equine encephalitis virus replicon particles (VRPs) significantly increased anti-PorB antibody levels. Boosting with rrPorB protein had little effect on antibody isotypes, while boosting with VRPs expressing PorB-enhanced a Th1 type response. Whole cell binding experiments showed that a portion of the antibodies recognized the surface of the homologous N. gonorrhoeae strain. Serum from groups with high antibody levels showed some opsonization of the homologous strain using human neutrophils. These results showed the potential of DNA vaccination for the purpose of priming an antibody response against PorB of N. gonorrhoeae. When combined with a protein or VRP boost, DNA priming resulted in high-titer and long-lasting responses. Based on different prime-boost protocols, we could polarize immune responses to predominantly Th1 or Th2, which should enable future studies of the types of immune responses that are protective in mouse models of gonorrhea. PMID- 14741159 TI - Differential antibody responses to a distinct region of human papillomavirus minor capsid proteins. AB - A peptide derived from the human papillomavirus type 16 (HPV-16) minor capsid protein, L2, has previously been reported to induce cross-neutralizing antibodies in mice. In this report, four HPV L2 peptides, including the HPV-16 peptide and its HPV type 6 and 11 homologues, along with extended peptides containing a conserved set of amino acids, were used to immunize rabbits and mice. Antibody responses were evaluated for specificity and ability to neutralize viral infection in vitro with a quantitative reverse transcriptase (RT)-polymerase chain reaction (PCR) assay. All peptide immunizations resulted in cognate and cross-peptide reactivity, but this did not translate equally into recognition of full-length protein, VLP, or neutralization of virus in vitro. This report provides the first evidence of cross-neutralization of authentic HPV by antiserum to L2 peptides. Comparison of the anti-peptide serum reactivity, especially with regard to neutralization of virus, indicates that the extended peptides may offer more potential to induce adequate responses for cross-protective immunity. PMID- 14741160 TI - Effect of congestive heart failure on humoral and ex vivo cellular immune responses to influenza vaccination in older adults. AB - This study examined the effect of congestive heart failure (CHF) on immune responses to influenza vaccination (2000-2001 preparation) in three groups of older adults including healthy, Class II and Class III/IV CHF. Serum antibody titers measured by hemagglutination inhibition (HI), and interferon-gamma (IFN gamma), interleukin-10 (IL-10) and granzyme B (GrzB) levels in ex vivo virus activated mononuclear cell cultures showed significant responses from pre vaccination to 4 and 12 weeks post-vaccination (P<0.01). There was a trend for lower GrzB and higher IFN-gamma and IL-10 levels in healthy versus CHF groups (P<0.06) for all viral strains at 4 weeks. HI titers did not differ between groups. In the regression model, Grz B levels were significantly predicted by the IFN-gamma:IL-10 ratio and performance on the 6 min Walk Test; age and CHF dropped out of the model. In conclusion, CHF in older adults predicts GrzB responses to influenza vaccination due to cytokine and physical ability differences. PMID- 14741161 TI - Modeling the economic net benefit of a potential vaccination program against ocular infection with Chlamydia trachomatis. AB - Trachoma is an ocular infection caused by Chlamydia trachomatis that carries a huge public health burden as a leading cause of preventable blindness globally. We developed a model of the economic benefits of a potential vaccine. Factors that impact the efficacy of a vaccination program (such as the sensitivity and specificity of algorithms to identify high-risk individuals), and personal and environmental risk factors for trachoma affect the maximum vaccination and screening costs at which a vaccination program can achieve a positive net benefit. This model is useful for planning vaccine programs in areas with varying trachoma risk factors and endemicity. PMID- 14741162 TI - Induction of CD8+ T cells to an HIV-1 antigen upon oral immunization of mice with a simian E1-deleted adenoviral vector. AB - An E1-deleted adenoviral recombinant derived from the chimpanzee serotype 6 expressing a codon-optimized truncated form of gag of human immunodeficiency virus type 1 (HIV-1) was tested for induction of a transgene product-specific CD8+ T cell response upon oral immunization of mice. The vector was shown to induce gag-specific CD8+ T cells detectable at moderate frequencies of approximately 0.5-1.0% in the spleens and to provide partial protection in a surrogate challenge model based on intraperitoneal (i.p.) infection of mice with a vaccinia virus recombinant expressing gag (VVgag) of HIV-1. Frequencies of gag specific CD8+ T cells could be augmented by using a different, i.e., heterologous, vaccine carrier based on a distinct recombinant virus or an alternative adenoviral serotype expressing the same form of gag for oral or systemic-booster immunization. PMID- 14741163 TI - Safety profile of recombinant canarypox HIV vaccines. AB - Attenuated poxviruses have been developed for use as candidate vaccine vectors. ALVAC, a strain of the Avipoxvirus canarypox, has been extensively evaluated as a vector for vaccines against the human immunodeficiency virus type 1 (HIV-1). This report presents the safety and reactogenicity data derived from 11 multicenter, randomized controlled trials of ALVAC-HIV vaccines conducted by the HIV Vaccine Trials Network (HVTN) and its predecessor, the AIDS Vaccine Evaluation Group (AVEG). Five different ALVAC vaccine constructs were tested among 1497 volunteers. Reactogenicity was similar for different ALVAC constructs. Local reactions of any grade to ALVAC vaccines were common. However, fewer than 2% of vaccinees had severe local responses, and less than 1% experienced severe local pain or tenderness. Systemic responses were mild and transient. As combination vaccine regimens are in common use, we also evaluated side effects of ALVAC vectors given in combination with a recombinant subunit protein. No significant differences were noted in the reactogenicity of ALVAC given with or without a recombinant envelope subunit vaccine. Black, non-Hispanic and male recipients of ALVAC-HIV reported less pain following vaccination than White, non-Hispanics and females, respectively. ALVAC-HIV vaccines are well tolerated at tested doses. The reactogenicity profiles are comparable to those reported for existing vaccines licensed for use among adults. Reactogenicity does not appear to be related to the number or type of inserted genes, and did not vary between different ALVAC constructs. PMID- 14741164 TI - Influenza virosomes enhance class I restricted CTL induction through CD4+ T cell activation. AB - Immunopotentiating reconstituted influenza virosomes (IRIV) are one of the few adjuvants currently licensed for human use. While their adjuvant capacity in the induction of humoral responses is clearly documented, few data exist on their effects on T cell immune response. Here we addressed IRIV adjuvance in the induction of HLA class I restricted cytotoxic T lymphocytes (CTL) in vitro. Lymphocyte stimulation with IM(58-66) and IRIV resulted in marked expansion of specific CTL as compared to cultures performed in the presence of either antigen alone or antigen and control liposomes (L). Studies addressing underlying adjuvant mechanisms demonstrated that IRIV activated CD4/CD45RO+ T cells, induced a cytokine profile consistent with T helper 1 (Th1) stimulation and increased the percentage of CD4+ T cells expressing CXCR3. Furthermore, supernatants from IRIV stimulated PBMC cultures promoted dendritic cell maturation. Most importantly, IRIV mediated CTL adjuvance required the presence of live CD4+ T cells. Powerful adjuvant effects of IRIV were also observed in the induction of CTL specific for the melanoma associated Melan-A/MART-1(27-35), HLA-A0201 restricted epitope. Taken together these findings indicate that IRIV are endowed with a high adjuvant capacity for HLA class I restricted CTL induction, largely attributable to their ability to antigenically stimulate CD4+ T cells. PMID- 14741165 TI - Characterization of M. Tuberculosis-derived IL-12-inducing material by alveolar macrophages. AB - We have investigated the substance derived from Mycobacterium tuberculosis (Mtb) that induces interleukin (IL)-12 production by alveolar macrophages (AMs) in vitro. The cytosol fraction of live Mtb H37Rv induced IL-12 production by AMs in a dose-dependent manner. The addition of interferon-gamma (IFN-gamma) augmented IL-12 production. IL-12-inducing activity by AMs (termed as surely active keeping rescue antigen, SAKRA) was purified by gel filtration and ion exchange column chromatography, and the molecular weight of SAKRA was estimated by gel filtration to be more than 700 kDa. SDS-polyacrylamide gel electrophoresis (PAGE) and Western blotting of SAKRA using rabbit anti-SAKRA antibody suggested that SAKRA is composed with several low molecular weight proteins. Amino acids sequence analysis of several bands after SDS-PAGE suggested that SAKRA is a part of ribosomes. RT-PCR showed that SAKRA induced not only expression of IL-12 p40 mRNA, but expression of tumor necrosis factor (TNF)-alpha and inducible nitric oxide synthase (iNOS) mRNA at least 6 h after stimulation, suggesting that SAKRA activates the bactericidal activity of macrophages. To investigate the potential use of SAKRA as a vaccine against tuberculosis, SAKRA was administered to BALB/c mouse that had been immunized with BCG for 18 months, and mouse were infected with Mtb H37Rv via a respiratory route. Replication of Mtb in lungs and spleens was examined 6 weeks after infection. Administration of SAKRA to BCG-vaccinated mice significantly reduced the numbers of Mtb in lungs and spleens as compared with BCG-vaccinated control mice. Taken together, these results suggest that SAKRA is one of the Mtb-derived immunomodulatory substances which induce IL-12 production during infection and also increases mycobactericidal activities of macrophages, and that SAKRA may be a promising new vaccine candidate against tuberculosis. PMID- 14741166 TI - Efficient delivery of DNA to dendritic cells mediated by influenza virosomes. AB - In an attempt to enhance the immunological efficacy of DNA-based vaccines, we have investigated a new biological means for delivering target gene DNA directly to professional antigen presenting cells (APC), such as the dendritic cells (DC), which are ultimately responsible for the antigen presentation and the primary activation of the immune system. For this purpose we investigated influenza virosomes (IRIV) with assembled DNA as a possible biological carrier for targeting the APC in vivo and in vitro. By cytofluorimetric analysis of the draining lymph nodes of Balb/c mice which had received (by intranasal (in.) administration) FITC-labeled DNA assembled with IRIV, we detected a significant labeled DNA uptake in a subset of lymph node deriving cells expressing DC surface markers. Subsequent mRNA analysis of these lymph nodes showed that the trans-gene delivered by the virosomes was effectively expressed as mRNA. Finally, a further cytofluorimetric analysis performed on human DC-enriched-PMBC, infected in vitro with labeled DNA/IRIV lead to the conclusion that the majority of APC (DC, B lymphocytes and CD16+ cells) are able to incorporate the labeled DNA transported by the construct. These findings suggest that the virosome is an efficient delivery system for testing infectious, as well as anti-cancer, DNA-based vaccine research. PMID- 14741167 TI - Adjuvant activity of Mycobacterium bovis BCG expressing CRM197 on the immune response induced by BCG expressing tetanus toxin fragment C. AB - In order to develop a combined recombinant Mycobacterium bovis BCG (rBCG) vaccine against diphtheria, pertussis and tetanus (DPT), we have constructed different strains of rBCG expressing tetanus toxin fragment C (FC), driven by the up regulated M. fortuitum beta-lactamase promoter, pBlaF*. Tetanus toxin FC was expressed in comparable levels in native form or in fusion with the beta lactamase exportation signal sequence; however, in both constructs it was localized to the cytosol. Immunization of mice with rBCG-FC or its combination with rBCG expressing CRM197, induced anti-tetanus toxin antibodies with a Th2 immunoglobulin profile. Administration of a subimmunizing dose of the diphtheria tetanus toxoid vaccine showed that rBCG-FC primed mice for production of an intense humoral response. Interestingly, the combination of rBCG-FC and rBCG CRM197 reduced the time required for maturation of the immune response and increased anti-tetanus toxin antibody levels, suggesting adjuvant properties for rBCG-CRM197; this combination induced 75% protection in mice challenged with 100 minimum lethal doses (MLD) of tetanus toxin. Antisera from guinea pigs immunized with this combination were shown to neutralize tetanus toxin and diphtheria toxin. Our results suggest reciprocal adjuvant effects of rBCG-FC and rBCG CRM197, which may contribute to induction of a more effective immune response against both diseases. PMID- 14741168 TI - Neutralization of primary HIV-1 SF13 can be detected in extended incubation phase assays with sera from monkeys immunized with recombinant HIV-1 SF2 gp120. AB - Phase III efficacy trials of recombinant human immunodeficiency virus type 1 (HIV 1) envelope glycoproteins were postponed. In Phase I and II trials these candidate vaccines had failed to induce neutralizing antibodies against virus which had been isolated by co-culture with human peripheral blood mononuclear cells (PBMC). The aim of the present study was to determine assay conditions for detecting neutralization of primary HIV-1 isolates with sera from immunized individuals. We show that in two immunogenicity trials in rhesus macaques, recombinant HIV-1 SF2 gp120 induced antibodies which neutralized the primary HIV 1 SF13 isolate. Statistically significant in vitro neutralization required assays in which the incubation phase was extended. Sterile immunity was only seen with the highest level of neutralization, induced by a recombinant prime, peptide boost strategy. We recommend that neutralization assays with extended incubation phases should be used to monitor Phase III efficacy trials. PMID- 14741169 TI - Comparison of different adjuvants of protein and DNA vaccination for the prophylaxis of IgE antibody formation. AB - A high-molecular-weight mite allergen Der f11 that was hardly purified for immunotherapy was used to develop the DNA vaccine pDf11. We have shown that vaccination of mice with pDf11 induces Th1 responses characterized by suppression of IgE responses. In the present study, effects of different adjuvants on pDf11 were first studied. Mice receiving pDf11 +/- CpG, bestatin, and bupivacaine had better suppression of IgE responses than those receiving pDf11 +/- lipofectin or alum. Bestatin could greatly boost IgG2a responses. Immunomodulating effects of different adjuvants between protein and DNA vaccines were further elucidated. CpG was the best for both protein and DNA vaccines to profoundly suppress IgE responses, but alum, bestatin and lipofectin were useless for rDf11 to induce IgE inhibition. Neither did the combination of rDf11 and pDf11 have further IgE suppression. In conclusion, CpG is the unique adjuvant for the protein vaccine rDf11 to inhibit IgE responses. In contrast, the DNA vaccine pDf11 +/- CpG, bestatin, or bupivacaine induces profound suppression of IgE responses. PMID- 14741170 TI - Prior DNA vaccination does not interfere with the live-attenuated measles vaccine. AB - The currently used live-attenuated measles vaccine is very effective although maternal antibody prevents its administration prior to 6 months of age. We are investigating the ability of a DNA vaccine encoding the measles viral hemagglutinin, fusion and nucleoprotein to protect newborn infants from measles. Here, we show that a measles DNA vaccine protects juvenile macaques from pathogenic measles virus challenge and that macaques primed and boosted with this DNA vaccine have anemnestic antibody and cell-mediated responses after vaccination with a live-attenuated canine distemper-measles vaccine. Therefore, this DNA vaccine administered to newborn infants may not hinder the subsequent use of live-attenuated measles vaccine. PMID- 14741171 TI - A post-licensure evaluation of the safety of inactivated hepatitis A vaccine (VAQTA, Merck) in children and adults. AB - BACKGROUND: Hepatitis A is a major cause of epidemic hepatitis in the US. In pre licensure trials, inactivated hepatitis A vaccine (HAV, VAQTA, Merck) was shown to be generally well-tolerated and effective in inducing immunity to hepatitis A infection in adults and children over 2 years of age. Following the licensure of this vaccine, we began a Phase IV safety evaluation in adults and in children over 2 years of age. METHODS: Safety was assessed by comparing the rates of diagnoses in clinic, emergency and hospital utilization. From April 1997 to December 1998, rates of diagnoses within 30 days for the clinic and emergency setting and 60 days for hospitalization were compared with unexposed follow-up time in the same individuals both before receipt of vaccine and after the 60 days interval post-vaccination. RESULTS: There were a total of approximately 2000 comparisons between the risk and "before" or "after" period. Among them, 106 were found to have statistically significant differences in rates (30 elevated, 76 lowered). Among children/adolescents (2-17 year-old), in the hospitalization category, the only statistically significant elevated risk found was "elective procedures", as compared with both "before" and "after" periods. In the outpatient visit category for children and adolescents, elevated risks were found for consultation/general medicine/exam when compared with both "before" and "after" periods, and ganglion and viral warts when compared with either "before" or "after" period. Among adults (> or =18 year-old), in the outpatient visit category, a statistically significant elevated relative risk was seen for diarrhea/gastroenteritis for both "before" and "after" periods. There were additionally 17 diagnostic categories that showed a statistically significantly elevated relative risk compared with either "before" or "after" period. Except for diarrhea/gastroenteritis, the other eight events were elevated only in one comparison (either "before" or "after"). These eight elevated relative risks might be explained by chance resulting from multiple comparison or seasonal variations. There were no serious adverse events judged by the investigator to be associated with HAV. CONCLUSION: In this large Phase IV evaluation of the safety of HAV, the vaccine appeared to be generally well-tolerated. These data support the continued routine use of HAV for vaccination in children and adults. PMID- 14741173 TI - Adolescent vaccination in the developing world: time for serious consideration? AB - The number of vaccinations and vaccine boosters that are becoming relevant to delivery among adolescents is growing. For sexually transmitted infections (STIs) in particular, such as HIV, vaccine delivery in this (pre-sexually active) age group is going to be an integral component of the strategy to control these diseases in the future. Currently, however, adolescent vaccination does not routinely occur because the traditional method of delivery is through secondary schools, and in developing countries school attendance has historically been low. But with school attendance growing rapidly throughout the developing world, the cost of vaccines falling over time, and an increasing number of current and future vaccines targeted at adolescents, it is an ideal opportunity to place this route of delivery on the national and international health agenda. Therefore, this paper examines the rationale for adolescent vaccination, looks at the success of other adolescent-targeted health care interventions, and finally, considers the challenges associated with future adolescent vaccination programmes in developing countries. PMID- 14741172 TI - Immunization of colorectal cancer patients with recombinant baculovirus-derived KSA (Ep-CAM) formulated with monophosphoryl lipid A in liposomal emulsion, with and without granulocyte-macrophage colony-stimulating factor. AB - KSA (Ep-CAM) is highly expressed by colorectal cancers. The safety and immunologic effects of a vaccine consisting of recombinant baculovirus-derived KSA formulated with monophosphoryl lipid A (MPL) in liposomes and emulsified in mineral oil were evaluated, with and without co-administration of granulocyte macrophage colony-stimulating factor (GM-CSF). Eleven patients with metastatic colorectal cancer received three subcutaneous (s.c.) injections of the vaccine at 4-week intervals. Six patients were randomized to also receive human recombinant GM-CSF (rGM-CSF) by subcutaneous injection daily for 4 days with each vaccination. Immunizations with and without rGM-CSF were well tolerated. Seven of the 11 patients developed significant KSA-specific cellular immune responses as assessed by lymphoproliferation and interferon-gamma (IFN-gamma) ELISPOT assays. All nine tested patients developed positive delayed type hypersensitivity reactions. Eight of the 11 patients developed KSA-specific antibody responses. The highest levels of cellular immune responses were observed in patients who received GM-CSF. Immunization with baculovirus-derived KSA formulated with monophosphoryl lipid A in liposomal emulsion is safe and can elicit KSA-specific immune responses. Co-administration of GM-CSF with this formulation is an effective method of generating KSA-specific T-helper (Th) 1-associated cellular immune responses. PMID- 14741174 TI - The introduction of new vaccines into developing countries. III. The role of intellectual property. AB - The development of new vaccines that address the particular needs of developing countries has been proceeding slowly. A number of new public sector vaccine research and development initiatives have been launched to address this problem. These new initiatives find that they often wish to collaborate with the private sector and, in collaborating with the private sector, they must address issues of intellectual property (IP) management. It has not been well understood why IP management is important and how such management by public sector groups can best be conducted. IP management has become very important because vaccine research and development is driven by the regulatory process. The regulatory process has increased the cost of vaccine development to very high levels especially for the highly sophisticated new vaccines currently under development. Thus, investors seek IP protection for the required large investments. Conversely, we assert this concept as a new insight, IP rights are essential for mobilizing the significant funds necessary to meet regulatory requirements. Thus, IP rights are of value not only for investors but also for the public at large. In the absence of public sector mechanisms to carry out the functions that the private sector currently conducts, the public sector needs to increase its sophistication in IP management and needs to identify and implement strategies that will help the public sector to achieve its public health goals, especially for the poor and, among these individuals, the poor in developing countries. This paper suggests some strategies that might be used by the public sector to help achieve its public health goals, especially for the poor. PMID- 14741175 TI - Sub-fragments of the envelope gene are highly protective against the Japanese encephalitis virus lethal infection in DNA priming--protein boosting immunization strategies. AB - The envelope (E) gene of Japanese encephalitis virus (JEV) plays a major protective role against JEV infection. In order to locate the part of E gene that is responsible for this protection, an N-terminal fragment EA (nucleotide number 933-1877 bp of JEV genome) and a C-terminal fragment EB (nucleotide number 1851 2330 bp of JEV genome) from E gene were prepared. Both of these fragments were used in the form of recombinant proteins (rEA and rEB) and plasmid DNA (pEA, pM15EA and pEB) for immunizations. Recombinant EA protein (rEA) was previously found to be non-protective because it was expressed in an insoluble form. Plasmid EA (pEA) was also found to be non-protective unless it is preceded by a 15 mer signal peptide derived from the very C-terminal of the membrane gene (M) of JEV to form pM15EA plasmid indicating the importance of the signal peptide in the expression of EA immunogenicity. Although pM15EA and pEB are both immunogenic and protective against JEV lethal infection, the protection by both fragments however is not optimal. Even when pM15EA and pEB were used together for immunization, maximum protection as those induced by control vaccine was not achieved. However, if individual fragments (EA or EB) were used in a DNA priming-protein boosting or protein priming-DNA boosting strategy, high levels of protection were achieved by both fragments. This was especially true for EA fragment where the level of protection against JEV lethal infection was equal to that induced by commercially available vaccine alone. The protection correlated very well with the neutralizing antibody titers and the T helper cell involved in this process in mainly the Th1 type. PMID- 14741176 TI - Parturient montes? PMID- 14741177 TI - A systematic review of the contribution of qualitative research to the study of quality of life in children and adolescents with epilepsy. AB - A sizeable literature focusing on QOL in children and adolescents with epilepsy has been produced over the last few years. However, relatively little emphasis has been placed on defining these issues from direct exploration of children's and adolescents' views. Qualitative methodologies are proposed in this review as an appropriate means of eliciting such information. This review systematically investigated the extent to which studies of QOL in children and adolescents with epilepsy have used recognised qualitative methodology. Articles for inclusion were identified by searching the term 'epilepsy', combined with 'adolescent(s) and/or child(ren)' and 'psychosocial and/or quality of life'. Selected articles were reviewed and rated using CASP Guidelines for qualitative research by two independent raters. Seventeen studies were retrieved through literature search. Of these six used some form of qualitative methodology either individually or combined with quantitative methods. However, only one study met quality criteria for selection in this systematic review. A summary of both selected and excluded studies is presented and methodological limitations discussed. Recommendations for appropriate methodology for investigation of QOL issues in children and adolescents are given. PMID- 14741178 TI - Quality of life and psychosocial development in adolescents with epilepsy: a qualitative investigation using focus group methods. AB - The majority of previous studies investigating the impact of epilepsy on the QOL of adolescents have used proxy opinions from clinicians and/or parents. This study highlights the need for research to investigate QOL from the direct perspective of adolescents and consider issues in the context of a developmental perspective. A focus group technique was used. Twenty-two adolescents aged between 12 years 4 months and 18 years 0 months (6 males and 16 females) were stratified by age (12-13, 14-15 and 16+ years) into six focus groups. Data were transcribed and QSR NUD*IST 4.0 was used to help generate central themes. Several procedures were undertaken to increase validity and reliability of findings. Analysis identified two main themes comprising (a) issues related to adolescent development (identity formation) and (b) epilepsy related variables, with five and four main sub-themes, respectively ('peer acceptance', 'development of autonomy', 'school related issues', 'epilepsy as part of me' and 'future', and 'medication issues', 'seizures', 'knowledge of epilepsy' and 'sense of uncertainty'). The main issues related to peer acceptance and development of autonomy. In contrast to previous studies, academic difficulties were not highlighted as an issue. No significant age-related differences in issues were identified. A conceptual model representing these findings is presented and clinical implications and suggestions for future research are reported. PMID- 14741179 TI - Video-EEG telemetry can be a crucial tool for neurologists experienced in epilepsy when diagnosing seizure disorders. AB - We retrospectively reviewed the charts of 121 patients consecutively admitted to our epilepsy-monitoring unit (VET) during the period of 01 July 2001 to 31 December 2002. We excluded patients with a confirmed diagnosis of epilepsy who were admitted for invasive pre-surgical monitoring. Medical records were reviewed to collect demographic and clinical information that lead to the initial referral for VET by neurologists with expertise in epilepsy or by an epileptologist. We identified 29 patients (24%), whose diagnosis changed after VET. Their seizure duration ranged from 1 to 46 years. A diagnosis of epileptic seizures (ES) was made in four of the patients who were initially felt to have nonepileptic seizures (NES). The diagnosis of NES was made in 22 patients who were initially felt to have ES. All of these 29 patients had failed at least two or more antiepileptic drugs (AEDs). A misclassification of epilepsy syndrome was found in three patients. Eleven of the NES patients had risk factors that would increase the likelihood of ES, including significant head injury (n=6), febrile seizures (n=2), meningioencephalitis (n=2), and tumours (n=1). Four of these 11 patients had abnormal interictal EEGs. We conclude that VET is crucial in establishing a diagnosis in patients with seizures. Without VET, patients can be misclassified or receive ineffective treatment, even when being treated by specialists in epilepsy. Thus, VET, can help facilitate the most appropriate type of therapy in difficult to control patients. PMID- 14741180 TI - Lateralisation value of lower limb behaviors in complex partial seizures of temporal lobe origin: a video-EEG analysis. AB - PURPOSE: To determine the lateralising value of leg behaviors in complex partial seizures (CPS) of temporal lobe onset. METHODS: Videotapes of 123 seizures from 38 patients who were seizure-free after temporal lobectomy were reviewed. Ictal behaviors including head turning, limb automatisms, tonic/dystonic postures and the latent time for ictal behavior were analysed for their lateralising value. RESULTS: Contralateral versive head turning, ipsilateral arm automatisms, contralateral arm tonic/dystonic posturing, and contralateral arm clonic posturing were found to have high predictive value of lateralisation. As for the lower limbs, meaningful leg behaviors were recorded in 38 (31%) of 123 seizures, far less than behaviors of the upper limbs (79%). The predictive value from leg behaviors were similar to that from upper limbs. Among the leg behaviors, dystonic behaviors were always contralateral to the ictal side. Tonic behaviors were 94% contralateral to the ictal side. Dystonia and clonic movement were always contralateral to the ictal side. Automatisms were 86% ipsilateral to the ictal side. CONCLUSIONS: Although incidences were low, leg behaviors could provide useful lateralising value of the seizure foci. Clinicians as well as investigators should recognize the value of lower limbs behavior in studies of ictal semiology. PMID- 14741181 TI - Aneurysms and epilepsy: an increasingly recognised cause. AB - Intracranial aneurysms have rarely been reported in association with complex partial seizures. We report five cases of complex partial seizures where aneurysms were found either accidentally or through investigations in these patients. MRI scans and angiograms revealed these aneurysms and in one patient several aneurysms were found. Four patients were treated with GDC coil embolisation with follow-on anti-epileptic medication. Two of the patients have reported a few minor seizures despite the treatment. The seizures in the other two patients have resolved completely after the treatment. The fifth patient is on anti-epileptic medication. We review the possible pathogenesis for the link between aneurysms and seizures. The treatment of intracranial aneurysms is also reviewed. PMID- 14741182 TI - Evidence-based standards of care for adults with epilepsy-a literature review. AB - Epilepsy is the most common serious chronic neurological condition. All patients with epilepsy regardless of location should receive the highest quality of services that can be provided. In order to do this current service provision has to be reviewed and bench marked against ideal standards of service provision that cover a comprehensive range of services to meet all patient needs. By critically reviewing the relevant literature, criteria were developed for an ideal epilepsy service. The literature review generated evidence-based ideal standards for the following service areas: - The role of primary care and the interface with secondary care in the management of epilepsy; - The role of Accident and Emergency (A and E) departments in the care of epilepsy; - Epilepsy clinics; - Specialist epilepsy nurses for treating epilepsy; - The role of the general practitioner (GP) specialist; - Services for adult epileptics with learning disabilities; - Services for complex epilepsy; and - User views. Although many of the recommendations are based on evidence of a lower grade, the direction of the existing evidence obtained from several sources, suggested similar standards for an epilepsy service. The effectiveness and value of epilepsy services, particularly in relation to the changes recommended above, must be further researched. PMID- 14741183 TI - Psychiatric adverse events in patients with epilepsy and learning disabilities taking levetiracetam. AB - PURPOSE: To investigate the prevalence and psychopathological features of psychiatric adverse events (PAEs) in patients with learning disabilities (LD) in therapy with levetiracetam (LEV). METHOD: From a population of 517 consecutively patients with epilepsy started on LEV, we identified 118 patients with epilepsy and LD. RESULTS: Fifteen patients (12.7%) experienced PAEs during LEV therapy. Two (1.7%) developed an affective disorder, nine (7.6%) aggressive behaviour, two (1.7%) emotion lability and two (1.7%) other personality changes such as agitation, anger and hostile behaviour. We observed a significant association with a previous history of status epilepticus and a previous psychiatric history. We did not find a statistically significant association with epilepsy diagnosis, age at onset or duration of the epilepsy, EEG or MRI features. The titration schedule of LEV appeared not to be relevant. CONCLUSIONS: LEV therapy was well tolerated in patients with epilepsy and LD and the main problems were related to aggressive behaviour. The titration schedule of LEV was not relevant and a subgroup of patients appeared to be biologically more vulnerable. PMID- 14741184 TI - Levetiracetam monotherapy for elderly patients with epilepsy. AB - We retrospectively identified 14 elderly patients with a history of partial seizures who received levetiracetam (LEV) monotherapy. Patients began LEV either as first line therapy (n=5) or were converted to LEV monotherapy (n=9) after failing prior antiepileptic medications (AEDs). Thirteen patients continued on LEV monotherapy for at least 6 months. One patient was lost to follow-up. Eight patients (61.5%) became seizure free. Four patients who began LEV as a first line therapy became seizure free, whereas the remaining four patients who converted to LEV after they failed their previous AEDs became seizure free. Four patients (30.7%) had more than a 50% seizure reduction of seizures. Only one patient had no significant change in seizure frequency after started on LEV. The total dosages used to control seizures were 500-3000 mg/day, (mean 1839.2 mg/day). LEV monotherapy can be effective and well tolerated in this group of patients. A prospective, larger, double blind monotherapy study is needed to confirm this finding. PMID- 14741185 TI - Thyroxine-induced hypermotor seizure. AB - Thyroxine-induced epilepsy is a very rare condition occurring in epileptic patients. Here we report a boy with thyroxine-induced hypermotor seizure (HMS) following thyroxine administration for his central hypothyroidism secondary to surgery and cranial radiation for his brain tumor. After 3 years seizure-free period, he had repeated HMS, seven to eight attacks per day, after initiation L thyroxine treatment. Following reduction of the daily thyroxine dose, his seizures decreased in frequency. To our knowledge, this is the first reported case of HMS associated with L-thyroxine administration. PMID- 14741187 TI - Molecular genetics of autosomal recessive polycystic kidney disease. AB - Autosomal recessive polycystic kidney disease (ARPKD) is a severe form of inherited childhood nephropathy ( approximately 1:20,000 live births) characterized by fusiform dilatation of collecting ducts and congenital hepatic fibrosis. Up to 30% die as neonates due to respiratory insufficiency and the majority of surviving infants develop hypertension. Progression to end stage renal disease occurs in 20-45% of cases within 15 years but a proportion maintain renal function into adulthood where complications of liver disease predominate. The ARPKD disease gene, PKHD1, has recently been identified through analysis of an orthologous animal model, the PCK rat. PKHD1 is a large gene ( approximately 470 kb) with 67 exons from which multiple transcripts may be generated by alternative splicing. It is highly expressed in kidney, with lower levels in liver and pancreas. The ARPKD protein, fibrocystin (4074 aa and 447 kDa), is predicted to be an integral membrane, receptor-like protein containing multiple copies of an Ig-like domain (TIG). Fibrocystin is localized to the branching ureteric bud, collecting and biliary ducts, consistent with the disease phenotype, and often absent from ARPKD tissue. In common with other PKD-related proteins, fibrocystin is localized to the primary cilia of renal epithelial cells, reinforcing the link between ciliary dysfunction and cyst development. Screens of PKHD1 have revealed 119 different mutations of various types spread throughout the gene. Several ancestral changes have been described, some localized to specific geographic populations. The majority of patients are compound heterozygotes and preliminary genotype/phenotype studies associate two truncating mutations with severe disease. The complexities of PKHD1, marked allelic heterogeneity and high level of missense changes complicate gene-based diagnostics. PMID- 14741188 TI - In vitro characterization of genetically modified embryonic stem cells as a therapy for murine mucopolysaccharidosis type IIIA. AB - The mucopolysaccharidoses (MPS) are lysosomal storage disorders resulting from the impaired catabolism of glycosaminoglycans (GAG). MPS type IIIA patients have dysfunctional sulfamidase enzyme leading to lysosomal storage of the GAG heparan sulfate, severe neurological symptoms including regression in learning, behavioural abnormalities, and premature death. We have engineered mouse D3 embryonic stem (ES) cells to over-express recombinant human sulfamidase. Human sulfamidase was correctly folded and secreted 2h post-labelling as determined by immunoprecipitation and SDS-PAGE analysis of transfected ES cells. Secreted human sulfamidase present in conditioned ES cell media was able to be taken up via mannose-6-phosphate-mediated endocytosis and restored sulfamidase enzyme activity in human MPS IIIA fibroblast cell lines. ES cells underwent directed differentiation to neural precursor populations and were capable of sustained human sulfamidase over-expression at all stages. Additionally, transfected and control cells were proliferative (Ki67+) and expressed several neural markers (nestin, MAP-2, and NF160) as determined by immunofluorescence. These findings suggest the possibility of ES cell-based therapy for the treatment of neurological pathology of MPS IIIA. PMID- 14741189 TI - Effects of odd-numbered medium-chain fatty acids on the accumulation of long chain 3-hydroxy-fatty acids in long-chain L-3-hydroxyacyl CoA dehydrogenase and mitochondrial trifunctional protein deficient skin fibroblasts. AB - The treatment for patients with genetic disorders of mitochondrial long-chain fatty acid beta-oxidation is directed toward providing sufficient sources of energy for normal growth and development, and at the same time preventing the adverse effects that precipitate or result from metabolic decompensation. Standard of care treatment has focused on preventing the mobilization of lipids that result from fasting and providing medium-chain triglycerides (MCT) in the diet in order to bypass the long-chain metabolic block. MCTs that are currently available as commercial preparations are in the form of even-chain fatty acids that are predominately a mixture of octanoate and decanoate. Recently, the use of odd-chain fatty acids has been proposed as an alternative treatment. We have shown previously that the even-numbered medium-chain fatty acids (MCFAs) that are found in MCT preparations can reduce the accumulation of potentially toxic long chain metabolites of fatty acid oxidation (FAO). In the current study, we undertook to determine if the same is true of odd-numbered MCFAs. We found that provision of odd-chain species does decrease the build-up of long-chain FAO intermediates in our in vitro skin fibroblast model, but to a lesser extent than even-numbered MCFAs. PMID- 14741190 TI - A novel deletion creating a new terminal exon of the dihydrolipoyl transacylase gene is a founder mutation of Filipino maple syrup urine disease. AB - Maple syrup urine disease (MSUD) is a rare, autosomal-recessive disorder of branched-chain amino-acid metabolism. In the Philippines, many MSUD cases have been diagnosed clinically. Here, molecular analysis of the dihydrolipoyl transacylase (E2) gene was done in 13 unrelated families from the Philippines. A novel deletion spanning 4.1 kb of intron 10 and 601 bp of exon 11, caused by non homologous recombination between an L1 repeat in intron 10 and an Alu repeat in exon 11, was found in 8 out of 13 families, with 5 of them being homozygous for the mutation, implicating it as a founder mutation of Filipino MSUD. The resulting mutant E2 mRNA contains a 239-bp insertion after exon 10, thereby producing a new terminal exon. Large-scale population screening of the deletion revealed that one carrier of the mutation was identified in 100 normal Filipinos. These findings suggest that a limited number of mutations might underlie MSUD in the Filipino population, potentially facilitating prenatal diagnosis and carrier detection of MSUD in this group. PMID- 14741191 TI - Galactitol and galactonate accumulation in heart and skeletal muscle of mice with deficiency of galactose-1-phosphate uridyltransferase. AB - Under conditions of dietary galactose loading, mice deficient in galactose-1 phosphate uridyltransferase (GALT) accumulate large amounts of galactitol and galactonate in heart and skeletal muscle. In contrast to liver, brain, and kidney, which form little galactitol when GALT-deficient animals (G/G) ingest a 40% galactose diet, heart and skeletal muscle galactitol reaches 22.90+/-1.62 (M+/-SE) and 38.88+/-2.62 micromol/g tissue, respectively, levels 40-100 times that of galactose-1-phosphate (Gal-1-P). Sixteen-day-old suckling G/G mice accumulate galactitol in heart and to a lesser extent, in skeletal muscle. Heart and skeletal muscle of G/G mice also form galactonate, with levels comparable to that of liver, which was presumed previously to be the only tissue capable of converting galactose to galactonate under conditions of loading. The data suggest that heart and skeletal muscle play a role in disposition of galactose when GALT activity is impaired, contributing a large share to urinary galactitol and galactonate excretion. The ability of heart and muscle to form galactonate may also contribute to the G/G mouse's ability to slowly oxidize galactose to CO2, since the compound is an intermediate in an alternate route for galactose disposition. PMID- 14741192 TI - Comparative genomic analysis of the HNF-4alpha transcription factor gene. AB - Hepatocyte nuclear factor-4alpha (HNF-4alpha), the gene for the maturity-onset diabetes of the young type 1 (MODY1) form of type 2 diabetes mellitus (T2DM), is within the T2DM-linked region on chromosome 20q12-q13.1 and consequently, is a positional candidate gene for T2DM. Mutations in the coding region of HNF-4alpha are rare in diabetes affected subjects. Altered regulation of HNF-4alpha gene expression, controlled by distant enhancer sequences, may contribute to the development of type 2 diabetes. Comparative sequence analysis was performed between 13 kb of genomic DNA 5' to the P1 promoter sequences of the human, mouse, and rat HNF-4alpha coding sequences. Three regions, located at -10.5 kb (295 bp in length), -6.25 kb (421 bp in length), and -5.36 kb (263 bp in length), have significant sequence identity between the species. These three regions were functionally characterized using the chloramphenicol acetyltransferase (CAT) reporter assay, in which the conserved 5' regions of mouse HNF-4alpha were cloned in front of the herpes simplex virus thymidine kinase promoter driving transcription of the CAT gene. A fragment containing the 421 bp conserved region significantly increased CAT activity in differentiated rat hepatoma cells (13.7 +/-1.9-fold control), while only a modest increase in CAT activity was observed in pancreatic cells (2.5-+/-0.9-fold control; 1.6-+/-0.1-fold control) and dedifferentiated hepatoma cells (1.7-+/-0.4-fold control). The remaining two conserved regions increased CAT activity minimally in pancreatic (1.1-+/-0.1-fold control to 1.9-+/-0.1-fold control) and hepatic (1.6-+/-0.5-fold control to 2.3 +/-0.4-fold control) cell lines. Denaturing high-performance liquid chromatography (DHPLC) was used to search for sequence variants in DNA from 259 T2DM individuals. Two single nucleotide polymorphisms (SNPs) were identified, both of which increased CAT activity in the insulinoma cell lines in the CAT reporter assay (1.4-fold increase over wild-type; 1.7-fold increase over wild type). These results suggest that comparative sequence analysis can efficiently identify regulatory elements and that sequence variants in regulatory elements of HNF-4alpha can contribute to altered HNF-4alpha gene expression. PMID- 14741193 TI - Association of the calpain-10 gene with type 2 diabetes mellitus in a Mexican population. AB - Variation in the calpain-10 gene (CAPN10) has been associated with risk of type 2 diabetes in the Mexican American population of Starr County, Texas. We typed five polymorphisms in the calpain-10 gene (SNP-43, -43, -63, and -110 and Indel-19) to test for association with type 2 diabetes in 248 individuals representative of the mestizo population of Mexico City and Orizaba, Mexico including 134 patients with type 2 diabetes and 114 subjects with normal fasting blood glucose levels. We found a significant difference in SNP-44 allele and genotype frequencies between type 2 diabetic and non-diabetic subjects. The rare allele at SNP-44 was associated with increased risk of type 2 diabetes (odds ratio (OR)=2.72, 95% confidence interval (CI)=1.16-6.35, P=0.017). SNP-110, which is in perfect linkage disequilibrium with SNP-44, was also associated with type 2 diabetes. The SNP-43, Indel-19, and SNP-63 haplogenotype 112/121 associated with significantly increased risk (OR=2.16, 95% CI=1.31-3.57) of type 2 diabetes in Mexican Americans was not associated with significantly increased in risk in Mexicans (OR=1.15, 95% CI=0.57-2.34). The results suggest that variation in CAPN10 affects risk of type 2 diabetes in the mestizo population of central Mexico (Mexico City and Orizaba) and in Mexican Americans (Starr County, Texas). PMID- 14741194 TI - Promoter haplotype combinations for the human PDGFRA gene are associated with risk of neural tube defects. AB - Recent animal studies suggested that deregulated expression of the platelet derived growth factor receptor alpha (PDGFRalpha) may contribute to the failure of normal neural tube closure (NTC). There is also suggestive evidence that the promoter haplotype of the PDGFRA is associated with genetic susceptibility in human neural tube defects (NTDs). The purpose of our study was to investigate the association between promoter haplotype combinations of the human PDGFRA gene and risk for NTDs in a Hispanic population from the Texas-Mexico border region. This population has a considerably higher prevalence of NTDs (16/10,000 live births) than that generally reported in the United States (8-10/10,000 live births). In the present study, NTDs were defined as spina bifida or anencephaly. The haplotype of PDGFRA gene promoter was determined by direct DNA sequence analysis. Two novel haplotypes, H2epsilon and H1beta, were found. We observed significant differences among variable haplotype groups from in vitro transient transfection studies in U2-OS osteosarcoma cell and two other cell lines (HeLa cell and MCF7 cell). Result from our case-control study demonstrated that the frequencies of haplotypes with low transcription activity were significantly higher in NTD mothers than that observed in control mothers (odds ratio=2.2, 95% CI=1.0-4.6). Infants with at least one low activity allele showed slightly higher risk (odds ratio=1.5, 95%=0.8-3.1). Our study suggests that the reduced transcriptional activity of PDGFRA gene could increase the risk of having an NTD-affected pregnancy. PMID- 14741195 TI - Identification of sequence variation in the galactose-1-phosphate uridyl transferase gene by dHPLC. AB - Transferase-deficient galactosaemia is an inherited disorder of carbohydrate metabolism, caused by mutation at the galactose-1-phosphate uridyl transferase (GALT) locus. A denaturing high performance liquid chromatography (dHPLC) method was developed for variant scanning of the GALT gene. The method unequivocally identified the Duarte D1, D2, Q188R, and K285N GALT alleles and associated polymorphisms. Length polymorphism in an intronic Alu repeat was characterised and a novel Single Nucleotide Polymorphism (IVS10nt-322g-->t) associated with the D1 allele was identified. PMID- 14741196 TI - Long-term follow-up of a patient with mild tetrahydrobiopterin-responsive phenylketonuria. AB - We report on the long-term follow-up of the first Italian patient with the tetrahydrobiopterin (BH4)-responsive type of phenylalanine hydroxylase deficiency (R243X/Y414C genotype). The patient was diagnosed by the newborn screening for phenylketonuria (PKU) and with a positive BH4 loading test. Introduction of BH4 (initially 10 and later 20 mg/kg/day) in addition to reduced low-phenylalanine diet resulted in therapeutic plasma phenylalanine concentrations (<340 micromol/L). Very good compliance and no side effects in this patient demonstrate the great potential of BH4 in the treatment of some patients with mild PKU. PMID- 14741197 TI - The c.419-420insA in the MTP gene is associated with abetalipoproteinemia among French-Canadians. AB - Abetalipoproteinemia (ABL) is a rare autosomal recessive disease characterised by the absence of apolipoprotein B (apoB) containing lipoproteins and, in consequence, very low triglyceride and cholesterol levels. Microsomal triglyceride transfer protein (MTP) has been associated with ABL. A search for sequence variants in the large subunit of MTP in a kindred of 10 individuals from Saguenay-Lac-St Jean area with a propositus exhibiting ABL as well as in four independent patients from the greater Quebec city area and exhibiting very low apoB and LDL-cholesterol levels identified 12 variations. Only one sequence variation, the c.419-420insA, was observed, in the homozygous form, in the abetalipoproteinemic patient. The -493G/-400A/-164T/282G/383T/419 420insA/453T/891C/969T/1151A/2884G haplotype carries the insertion and was found in all members of the family studied. In conclusion, the present study showed that the c.419-420insA alone, in the homozygous form, is a cause of classical recessive inherited ABL in the French-Canadian population. PMID- 14741198 TI - Cerebrotendinous xanthomatosis in a Hong Kong Chinese kinship with a novel splicing site mutation IVS6-1G>T in the sterol 27-hydroxylase gene. AB - We reported a Hong Kong Chinese proband with Cerebrotendinous Xanthomatosis in which a novel acceptor splicing site mutation (IVS6-1G>T) was identified. Family screening revealed the same mutation in his elder brother and the youngest sister. All the three affected siblings were compound heterozygous for IVS6-1G>T and a known missense mutation R372Q (GenBank Accession No. M62401). Significant phenotypic variation was noted among them that the youngest sister was still symptom-free at the time of writing. PMID- 14741199 TI - Fast folding of the HIV-1 and SIV gp41 six-helix bundles. AB - Human (HIV-1) and simian (SIV) immunodeficiency virus fusion with the host cell is promoted by the receptor-triggered refolding of the gp41 envelope protein into a stable trimer-of-hairpins structure that brings viral and cellular membranes into close proximity. The core of this hairpin structure is a six-helix bundle in which an inner homotrimeric coiled coil is buttressed by three antiparallel outer HR2 helices. We have used stopped-flow circular dichroism spectroscopy to characterize the unfolding and refolding kinetics of the six-helix bundle using the HIV-1 and SIV N34(L6)C28 polypeptides. In each case, the time-course of ellipticity changes in refolding experiments is well described by a simple two state model involving the native trimer and the unfolded monomers. The unfolding free energy of the HIV-1 and SIV trimers and their urea dependence calculated from kinetic data are in very good agreement with data measured directly by isothermal unfolding experiments. Thus, formation of the gp41 six-helix bundle structure involves no detectable population of stable, partly folded intermediates. Folding of HIV-1 N34(L6)C28 is five orders of magnitudes faster than folding of its SIV counterpart in aqueous buffer: k(on),(HIV-1)=1.3 x 10(15)M(-2)s(-1) versus k(on),(SIV)=1.1 x 10(10)M(-2)s(-1). The unfolding rates are similar: k(off),(HIV-1)=1.1 x 10(-5)s(-1) versus k(off),(SIV=)5.7 x 10(-4)s( 1). Kinetic m-values indicate that the transition state for folding of the HIV-1 protein is significantly more compact than the transition state of the SIV protein. Replacement of a single SIV threonine by isoleucine corresponding to position 573 in the HIV-1 sequence significantly stabilizes the protein and renders the folding rate close to that of the HIV-1 protein yet without making the transition state of the mutant as compact as that of the HIV-1 protein. Therefore, the overall reduction of surface exposure in the high-energy transition state seems not to account for different folding rates. While the available biological evidence suggests that refolding of the gp41 protein is slow, our study implies that structural elements outside the trimer-of-hairpins limit the rate of HIV-1 fusion kinetics. PMID- 14741200 TI - A cross-talk between hypoxia and TGF-beta orchestrates erythropoietin gene regulation through SP1 and Smads. AB - Erythropoietin (Epo) is the humoral regulator of red blood-cell production. Low oxygen tension increases the Epo levels by enhancing transcription, through the hypoxia-inducible factor (HIF)-1, a transcriptional modulator in oxygen-regulated gene expression. In the present work, a cooperative interaction between hypoxia, mediated by the HIF-1 complex, and transforming growth factor-beta (TGF-beta), mediated by Smad3/4, was revealed in the Epo gene. This cooperation is due to physical interaction between Smad3/4 and HIF-1alpha. The Smad3/4 binding site is located within the 3' Epo enhancer, downstream from the HRE consensus, and immediately adjacent to the orphan hepatic nuclear factor receptor (HNF-4). HNF-4 is interacting also with Smad3 and the HIF-1 complex, to potentiate further the cooperative effect between both factors. Moreover, Sp1 has been identified as the factor binding the promoter necessary for the full hypoxia inducibility of the EPO gene. However, this full induction is achieved only if the TGF-beta pathway is mediating a cross-talk between promoter (Sp1) and enhancer (HIF-1alpha) regions through Smad3. We show that Sp1 binding to the proximal promoter is relevant for Epo transcription, and contributes to the Epo induction by hypoxia. A functional cooperation among the transcription factors mediating hypoxia (HIF 1, Sp1), the TGF-beta pathway (Smad3/4), and tissue-specific HNF-4 is proposed for the regulation of the Epo gene. In this model, the physical contact between the upstream promoter and the 3' downstream enhancer is mediated by Sp1 and Smad3 factors, and would occur upon bending of the DNA intervening sequences. Thus, Sp1 would reinforce the promoter/enhancer contact, while Smad3 would stabilize the multifactorial complex by interacting with HIF-1/Sp1/HNF-4 and the coactivator CBP/p300. This model may be extended to other genes where collaboration between TGF-beta and hypoxia takes place. PMID- 14741201 TI - Purine and pyrimidine-specific repression of the Escherichia coli carAB operon are functionally and structurally coupled. AB - Transcription of the carAB operon encoding the sole carbamoylphosphate synthetase of Escherichia coli proceeds from a tandem pair of promoters. P2, downstream, is repressed by arginine and the ArgR protein, whereas P1 is submitted to pyrimidine specific regulation and as shown here to purine-specific control exerted by binding of the PurR protein to a PUR box sequence centered around nucleotide 128.5 with respect to the start of P1 transcription. In vivo analyses of the effects of trans and cis-acting mutations on the regulatory responses and single round in vitro transcription assays indicated that ligand-bound PurR is by itself unable to inhibit P1 promoter activity. To exert its effect PurR relies on the elaborated nucleoprotein complex that governs P1 activity in a pyrimidine specific manner. Thus we reveal the existence of an unprecedented functional and structural coupling between the modulation of P1 activity by purine and pyrimidine residues that appears to result from the unique position of the PUR box in the carAB control region, far upstream of the promoter. Missing contact and premethylation binding interference studies revealed the importance of base specific groups and of structural aspects of the PUR box sequence in complex formation. Permutation assays indicated that the overall PurR-induced bending of the carAB control region is slightly less pronounced than that of the purF operator. The PUR boxes of the carAB operon of E.coli and Salmonella typhimurium are unique in that they have a guanine residue at position eight. Interestingly, guanine at this position has been proposed to be extremely unfavorable on the basis of modeling and binding studies, as its exocyclic amino group would enter into a steric clash with the side-chain of lysine 55. To analyze the effect of guanine at position eight in the upstream half-site of the carAB operator we constructed the adenine derivative and assayed in vivo repressibility of P1 promoter activity and in vitroPurR binding to the mutant operator, and constructed a molecular model for the unusual lysine 55-guanine 8 interaction. PMID- 14741202 TI - Mechanisms of homogeneous nucleation of polymers of sickle cell anemia hemoglobin in deoxy state. AB - The primary pathogenic event of sickle cell anemia is the polymerization of the mutant hemoglobin (Hb) S within the red blood cells, occurring when HbS is in deoxy state in the venous circulation. Polymerization is known to start with nucleation of individual polymer fibers, followed by growth and branching via secondary nucleation, yet the mechanisms of nucleation of the primary fibers have never been subjected to dedicated tests. We implement a technique for direct determination of rates and induction times of primary nucleation of HbS fibers, based on detection of emerging HbS polymers using optical differential interference contrast microscopy after laser photolysis of CO-HbS. We show that: (i). nucleation throughout these determinations occurs homogeneously and not on foreign substrates; (ii). individual nucleation events are independent of each other; (iii). the nucleation rates are of the order of 10(6)-10(8)cm(-3)s(-1); (iv). nucleation induction times agree with an a priori prediction based on Zeldovich's theory; (v). in the probed parameter space, the nucleus contains 11 or 12 molecules. The nucleation rate values are comparable to those leading to erythrocyte sickling in vivo and suggest that the mechanisms deduced from in vitro experiments might provide physiologically relevant insights. While the statistics and dynamics of nucleation suggest mechanisms akin to those for small molecule and protein crystals, the nucleation rate values are nine to ten orders of magnitude higher than those known for protein crystals. These high values cannot be rationalized within the current understanding of the nucleation processes. PMID- 14741203 TI - Negative effect of genetic bottlenecks on the adaptability of vesicular stomatitis virus. AB - Muller's ratchet is a principle of evolutionary genetics describing mutant accumulation in populations that are repeatedly subjected to genetic bottleneck. The immediate effect of Muller's ratchet, overall loss of fitness, has been confirmed in several viral systems belonging to different groups. This report shows that in addition to fitness loss, genetic bottlenecks also have longer-term effects, namely changes in the capacity of viral populations to adapt. Thus, vesicular stomatitis virus strains with a history of genetic bottleneck have lower adaptability than strains maintained at relatively large population sizes. This lower adaptability is illustrated by their reduced ability to regain fitness and by their inability to outcompete wild-type populations in situations where the initial fitness of the bottlenecked mutant is the same or even higher than the initial fitness of the wild-type. PMID- 14741204 TI - Conformational flexibility of four-way junctions in RNA. AB - Helical junctions are common architectural features in RNA. They are particularly important in autonomously folding molecules, as exemplified by the hairpin ribozyme. We have used single-molecule fluorescence spectroscopy to study the dynamic properties of the perfect (4H) four-way helical junction derived from the hairpin ribozyme. In the presence of Mg(2+), the junction samples parallel and antiparallel conformations and both stacking conformers, with a bias towards one antiparallel stacking conformer. There is continual interconversion between the forms, such that there are several transitions per second under physiological conditions. Our data suggest that interconversion proceeds via an open intermediate with reduced cation binding in which coaxial stacking between helices is disrupted. The rate of interconversion becomes slower at higher Mg(2+) concentrations, yet the activation barrier decreases under these conditions, indicating that entropic effects are important. Transitions also occur in the presence of Na(+) only; however, the coaxial stacking appears incomplete under these conditions. The polymorphic and dynamic character of the four-way RNA junction provides a source of structural diversity, from which particular conformations required for biological function might be stabilised by additional RNA interactions or protein binding. PMID- 14741205 TI - Generation of the BfiI restriction endonuclease from the fusion of a DNA recognition domain to a non-specific nuclease from the phospholipase D superfamily. AB - The BfiI endonuclease cleaves DNA at fixed positions downstream of an asymmetric sequence. Unlike other restriction enzymes, it functions without metal ions. The N-terminal half of BfiI is similar to Nuc, an EDTA-resistant nuclease from Salmonella typhimurium that belongs to the phosphoplipase D superfamily. Nuc is a dimer with one active site at its subunit interface, as is BfiI, but it cuts DNA non-specifically. BfiI was cleaved by thermolysin into an N-terminal domain, which forms a dimer with non-specific nuclease activity, and a C-terminal domain, which lacks catalytic activity but binds specifically to the recognition sequence as a monomer. On denaturation with guanidinium, BfiI underwent two unfolding transitions: one at a relatively low concentration of guanidinium, to a dimeric non-specific nuclease; a second at a higher concentration, to an inactive monomer. The isolated C-terminal domain unfolded at the first (relatively low) concentration, the isolated N-terminal at the second. Hence, BfiI consists of two physically separate domains, with catalytic and dimerisation functions in the N terminus and DNA recognition functions in the C terminus. It is the first example of a restriction enzyme generated by the evolutionary fusion of a DNA recognition domain to a phosphodiesterase from the phospholipase D superfamily. BfiI may consist of three structural units: a stable central core with the active site, made from two copies of the N-terminal domain, flanked by relatively unstable C terminal domains, that each bind a copy of the recognition sequence. PMID- 14741206 TI - Regions within the N-terminal domain of human topoisomerase I exert important functions during strand rotation and DNA binding. AB - The human topoisomerase I N-terminal domain is the only part of the enzyme still not crystallized and the function of this domain remains enigmatical. In the present study, we have addressed the specific functions of individual N-terminal regions of topoisomerase I by characterizing mutants lacking amino acid residues 1-202 or 191-206 or having tryptophane-205 substituted by glycine in a broad variety of in vitro activity assays. As a result of these investigations we find that mutants altered in the region 191-206 distinguished themselves from the wild type enzyme by a faster strand rotation step, insensitivity towards the anti cancer drug camptothecin in relaxation and the inability to ligate blunt end DNA fragments. The mutant lacking amino acid residues 1-202 was impaired in blunt end DNA ligation and showed wild-type sensitivity towards camptothecin in relaxation. Taken together, the presented data support a model according to which tryptophane 205 and possibly other residues located between position 191-206 coordinates the restriction of free strand rotation during the topoisomerization step of catalysis. Moreover, tryptophane-205 appears important for the function of the bulk part of the N-terminal domain in direct DNA interaction. PMID- 14741207 TI - The 40 kDa carboxy-terminal domain of poly(ADP-ribose) polymerase-1 forms catalytically competent homo- and heterodimers in the absence of DNA. AB - The 40 kDa carboxy-terminal catalytic domain (CD) of avian poly(ADP-ribose) polymerase (PARP-1) was cloned, expressed in a baculovirus expression system, and purified to homogeneity by affinity chromatography. The purified polypeptide synthesized covalent CD-poly(ADP-ribose) conjugates in the absence of DNA. Electrophoretic analysis of the ADP-ribose chain length distribution generated indicated that recombinant CD was able to catalyze the initiation, elongation, and branching reactions of poly(ADP-ribose) synthesis, although at a 500-fold lower efficiency than wild-type PARP-1. Kinetic evaluation of poly(ADP-ribose) synthesis showed that the enzymatic activities of CD increased for up to 60 minutes in a time-dependent manner. Moreover, the rates of CD auto-poly(ADP ribosyl)ation increased with second-order kinetics as a function of the protein concentration with either betaNAD(+) or 3'-deoxyNAD(+) as a substrate. Furthermore, the formation of catalytically competent CD-[PARP-1] heterodimers was also observed in specific ultrafiltration experiments. Thus, we conclude that the 40 kDa carboxy terminus of PARP-1 forms a competent catalytic dimer in the absence of DNA, and that its automodification reaction is intermolecular. PMID- 14741208 TI - Mining the structural genomics pipeline: identification of protein properties that affect high-throughput experimental analysis. AB - Structural genomics projects represent major undertakings that will change our understanding of proteins. They generate unique datasets that, for the first time, present a standardized view of proteins in terms of their physical and chemical properties. By analyzing these datasets here, we are able to discover correlations between a protein's characteristics and its progress through each stage of the structural genomics pipeline, from cloning, expression, purification, and ultimately to structural determination. First, we use tree based analyses (decision trees and random forest algorithms) to discover the most significant protein features that influence a protein's amenability to high throughput experimentation. Based on this, we identify potential bottlenecks in various stages of the structural genomics process through specialized "pipeline schematics". We find that the properties of a protein that are most significant are: (i.) whether it is conserved across many organisms; (ii). the percentage composition of charged residues; (iii). the occurrence of hydrophobic patches; (iv). the number of binding partners it has; and (v). its length. Conversely, a number of other properties that might have been thought to be important, such as nuclear localization signals, are not significant. Thus, using our tree-based analyses, we are able to identify combinations of features that best differentiate the small group of proteins for which a structure has been determined from all the currently selected targets. This information may prove useful in optimizing high-throughput experimentation. Further information is available from http://mining.nesg.org/. PMID- 14741209 TI - The structure of Escherichia coli ATP-phosphoribosyltransferase: identification of substrate binding sites and mode of AMP inhibition. AB - ATP-phosphoribosyltransferase (ATP-PRT), the first enzyme of the histidine pathway, is a complex allosterically regulated enzyme, which controls the flow of intermediates through this biosynthetic pathway. The crystal structures of Escherichia coli ATP-PRT have been solved in complex with the inhibitor AMP at 2.7A and with product PR-ATP at 2.9A (the ribosyl-triphosphate could not be resolved). On the basis of binding of AMP and PR-ATP and comparison with type I PRTs, the PRPP and parts of the ATP-binding site are identified. These structures clearly identify the AMP as binding in the 5-phosphoribosyl-alpha-1-pyrophosphate (PRPP)-binding site, with the adenosine ring occupying the ATP-binding site. Comparison with the recently solved Mycobacterium tuberculosis ATP-PRT structures indicates that histidine is solely responsible for the large conformational changes observed between the hexameric forms of the enzyme. The role of oligomerisation in inhibition and the structural basis for the synergistic inhibition by histidine and AMP are discussed. PMID- 14741210 TI - Induction and myofibrillar targeting of CARP, and suppression of the Nkx2.5 pathway in the MDM mouse with impaired titin-based signaling. AB - Muscular dystrophy with myositis (mdm) is a recessive mouse mutation that is caused by a small deletion in the giant elastic muscle protein titin. Homozygous mdm/mdm mice develop a progressive muscular dystrophy, leading to death at approximately 2 months of age. We surveyed the transcriptomes of skeletal muscles from 24 day old homozygous mdm/mdm and +/+ wild-type mice, an age when MDM animals have normal passive and active tensions and sarcomeric structure. Of the 12488 genes surveyed (U74 affymetrix array), 75 genes were twofold to 30-fold differentially expressed, including CARP (cardiac ankyrin repeat protein), ankrd2/Arpp (a CARP-like protein) and MLP (muscle LIM protein), all of which associate with the titin filament system. The four genes most strongly affected (eightfold to 30-fold change) were all members of the CARP-regulated Nkx-2.5 dependent signal pathway, and CARP mRNA level was 30-fold elevated in MDM skeletal muscle tissues. The CARP protein overexpressed in MDM became associated with the I-band region of the sarcomere. The mdm mutation excises the C-terminal portion of titin's N2A region, abolishing its interaction with p94/calpain-3 protease. Thus, the composition of the titin N2A protein complex is altered in MDM by incorporation of CARP and loss of p94/calpain-3. These changes were absent from the following control tissues (1). cardiac muscles from homozygous mdm/mdm animals, (2). skeletal and cardiac muscle from heterozygous mdm/+ animals, and (3). dystrophic muscles from MDX mice. Thus, the altered composition of the titin N2A complex is specific for the titin-based skeletal muscular dystrophy in MDM. PMID- 14741211 TI - Inositol 1,4,5-trisphosphate receptor contains multiple cavities and L-shaped ligand-binding domains. AB - Calcium concentrations are strictly regulated in all biological cells, and one of the key molecules responsible for this regulation is the inositol 1,4,5 trisphosphate receptor, which was known to form a homotetrameric Ca(2+) channel in the endoplasmic reticulum. The receptor is involved in neuronal transmission via Ca(2+) signaling and for many other functions that relate to morphological and physiological processes in living organisms. We analysed the three dimensional structure of the ligand-free form of the receptor based on a single particle technique using an originally developed electron microscope equipped with a helium-cooled specimen stage and an automatic particle picking system. We propose a model that explains the complex mechanism for the regulation of Ca(2+) release by co-agonists, Ca(2+), inositol 1,4,5-trisphosphate based on the structure of multiple internal cavities and a porous balloon-shaped cytoplasmic domain containing a prominent L-shaped density which was assigned by the X-ray structure of the inositol 1,4,5-trisphosphate binding domain. PMID- 14741212 TI - Cystatin forms a tetramer through structural rearrangement of domain-swapped dimers prior to amyloidogenesis. AB - The cystatins were the first amyloidogenic proteins to be shown to oligomerize through a 3D domain swapping mechanism. Here we show that, under conditions leading to the formation of amyloid deposits, the domain-swapped dimer of chicken cystatin further oligomerizes to a tetramer, prior to fibrillization. The tetramer has a very similar circular dichroism and fluorescence signature to the folded monomer and dimer structures, but exhibits some loss of dispersion in the 1H-NMR spectrum. 8-Anilino-1-naphthalene sulfonate fluorescence enhancement indicates an increase in the degree of disorder. While the dimerization reaction is bimolecular and most likely limited by the availability of a predominantly unfolded form of the monomer, the tetramerization reaction is first-order. The tetramer is formed slowly (t(1/2)=six days at 85 degrees C), dimeric cystatin is the precursor to tetramer formation, and thus the rate is limited by structural rearrangement within the dimer. Some higher-order oligomerization events parallel tetramer formation while others follow from the tetrameric form. Thus, the tetramer is a transient intermediate within the pathway of large-scale oligomerization. PMID- 14741213 TI - The ordering of corneal collagen fibrils with increasing ionic strength. AB - The fixed stromal charge of bovine corneas, osmotically clamped at physiological hydration, was altered by regulating the amount of chloride ions bound to the matrix. We measured the local fibrillar collagen order using X-ray diffraction methods. As the bound anions increased up to physiological values, the local fibrillar order increased to an optimal value. The coherence distance (t) approximately doubles to a maximum value (409 nm) from 10 mM NaCl to 154 mM NaCl. This then slowly decreased as the bathing solution increased to 1000 mM. In contrast the diameter of the collagen fibrils were minimal at physiological NaCl. PMID- 14741214 TI - Structural distortion of p53 by the mutation R249S and its rescue by a designed peptide: implications for "mutant conformation". AB - Missense mutations in the DNA-binding core domain of the tumour suppressor protein p53 are frequent in cancer. Many of them result in loss of native structure. The mutation R249S is one of the six most common cancer-associated p53 mutations ("hot-spots"). As it is highly frequent in hepatocellular carcinoma, its rescue is an important therapeutic target. We have used NMR techniques to study the structural effects of the R249S mutation. The overall fold of the core domain is retained in R249S, and it does not take up a denatured "mutant conformation". However, the beta-sandwich had increased flexibility and, according to changes in chemical shift, there was local distortion throughout the DNA-binding interface. It is likely that the R249S mutation resulted in an ensemble of native and native-like conformations in a dynamic equilibrium. The peptide FL-CDB3 that was designed to rescue mutants of p53 by binding specifically to its native structure was found to revert the chemical shifts of R249S back towards the wild-type values and so reverse the structural effects of mutation. We discuss the implications for a rescue strategy and also for the analysis of antibody-binding data. PMID- 14741215 TI - Binding of p53-derived ligands to MDM2 induces a variety of long range conformational changes. AB - We have used NMR to study the effects of peptide binding on the N-terminal p53 binding domain of human MDM2 (residues 25-109). There were changes in HSQC chemical shifts throughout the domain on binding four different p53-derived peptide ligands that were significantly large to be indicative of global conformational changes. Large changes in chemical shift were observed in two main regions: the peptide-binding cleft that directly binds the p53 ligands; and the hinge regions connecting the beta-sheet and alpha-helical structures that form the binding cleft. These conformational changes reflect the adaptation of the cleft on binding peptide ligands that differ in length and amino acid composition. Different ligands may induce different conformational transitions in MDM2 that could be responsible for its function. The dynamic nature of MDM2 might be important in the design of anti-cancer drugs that are targeted to its p53 binding site. PMID- 14741216 TI - Protein stability in nanocages: a novel approach for influencing protein stability by molecular confinement. AB - Confinement of a protein in a small inert space and microviscosity are known to increase its thermodynamic stability in a way similar to the mechanisms that stabilize protein fold in the cell. Here, to examine the influence of confinement on protein stability we choose four test cases of single domain proteins characterized by a wide range of melting temperatures, from approximately 73 degrees C of titin I27 to approximately 36 degrees C of yeast frataxin. All proteins are stabilized when confined in the gel, the most dramatic stabilization being that of yeast frataxin, whose melting temperature increased by almost 5 degrees C in the gel. In addition to being simple to use, this approach allows us to change the viscosity of the solvent without changing its composition or altering the structure of the proteins. The dimensions of the pores of the gels fall in the nanometer range, hence they are similar to those of the chaperone cavity. This method could therefore be used as a novel and powerful approach for protein folding studies. PMID- 14741217 TI - Crystal structure of the catalytic domain of MMP-16/MT3-MMP: characterization of MT-MMP specific features. AB - Membrane-type matrix metalloproteinases (MT-MMPs) have attracted strong attention, because four of them can activate a key player in the tumor scenario, proMMP-2/progelatinase A. In addition to this indirect effect on the cellular environment, these MT-MMPs degrade extracellular matrix proteins, and their overproduction is associated with tumor growth. We have solved the structure of the catalytic domain (cd) of MT3-MMP/MMP-16 in complex with the hydroxamic acid inhibitor batimastat. CdMT3-MMP exhibits a classical MMP-fold with similarity to MT1-MMP. Nevertheless, it also shows unique properties such as a modified MT specific loop and a closed S1' specificity pocket, which might help to design specific inhibitors. Some MT-MMP-specific features, derived from the crystal structures of MT-1-MMP determined previously and MT3-MMP, and revealed in recent mutagenesis experiments, explain the impaired interaction of the MT-MMPs with TIMP-1. Docking experiments with proMMP-2 show some exposed loops including the MT-loop of cdMT3-MMP involved in the interaction with the proMMP-2 prodomain in the activation encounter complex. This model might help to understand the experimentally proven importance of the MT-loop for the activation of proMMP-2. PMID- 14741218 TI - Folding and association of an extremely stable dimeric protein from Sulfolobus islandicus. AB - ORF56 is a plasmid-encoded protein from Sulfolobus islandicus, which probably controls the copy number of the pRN1 plasmid by binding to its own promotor. The protein showed an extremely high stability in denaturant, heat, and pH-induced unfolding transitions, which can be well described by a two-state reaction between native dimers and unfolded monomers. The homodimeric character of native ORF56 was confirmed by analytical ultracentrifugation. Far-UV circular dichroism and fluorescence spectroscopy gave superimposable denaturant-induced unfolding transitions and the midpoints of both heat as well as denaturant-induced unfolding depend on the protein concentration supporting the two-state model. This model was confirmed by GdmSCN-induced unfolding monitored by heteronuclear 2D NMR spectroscopy. Chemical denaturation was accomplished by GdmCl and GdmSCN, revealing a Gibbs free energy of stabilization of -85.1 kJ/mol at 25 degrees C. Thermal unfolding was possible only above 1 M GdmCl, which shifted the melting temperature (t(m)) below the boiling point of water. Linear extrapolation of t(m) to 0 M GdmCl yielded a t(m) of 107.5 degrees C (5 microM monomer concentration). Additionally, ORF56 remains natively structured over a remarkable pH range from pH 2 to pH 12. Folding kinetics were followed by far-UV CD and fluorescence after either stopped-flow or manual mixing. All kinetic traces showed only a single phase and the two probes revealed coincident folding rates (k(f), k(u)), indicating the absence of intermediates. Apparent first-order refolding rates depend linearly on the protein concentration, whereas the unfolding rates do not. Both lnk(f) and lnk(u) depend linearly on the GdmCl concentration. Together, folding and association of homodimeric ORF56 are concurrent events. In the absence of denaturant ORF56 refolds fast (7.0 x 10(7)M(-1)s(-1)) and unfolds extremely slowly (5.7 year(-1)). Therefore, high stability is coupled to a slow unfolding rate, which is often observed for proteins of extremophilic organisms. PMID- 14741219 TI - Heterogeneous folding of the trpzip hairpin: full atom simulation and experiment. AB - The beta-hairpin trpzip2 can be tuned continuously from a two-state folder to folding on a rough energy landscape without a dominant refolding barrier. At high denaturant concentration, this extremely stable peptide exhibits a single apparent "two-state" transition temperature when monitored by different spectroscopic techniques. However, under optimal folding conditions the hairpin undergoes an unusual folding process with three clusters of melting transitions ranging from 15 degrees C to 160 degrees C, as monitored by 12 different experimental and computational observables. We explain this behavior in terms of a rough free energy landscape of the unfolded peptide caused by multiple tryptophan interactions and alternative backbone conformations. The landscape is mapped out by potentials of mean force derived from replica-exchange molecular dynamics simulations. Implications for deducing cooperativity from denaturant titrations, for the origin of folding cooperativity, and for the folding of thermophilic proteins are pointed out. trpzip is an excellent small tunable model system for the glass-like folding transitions predicted by landscape theory. PMID- 14741220 TI - The regions of the sequence most exposed to the solvent within the amyloidogenic state of a protein initiate the aggregation process. AB - Formation of misfolded aggregates is an essential part of what proteins can do. The process of protein aggregation is central to many human diseases and any aggregating event needs to be prevented within a cell and in protein design. In order to aggregate, a protein needs to unfold its native state, at least partially. The conformational state that is prone to aggregate is difficult to study, due to its aggregating potential and heterogeneous nature. Here, we use a systematic approach of limited proteolysis, in combination with electrospray ionisation mass spectrometry, to investigate the regions that are most flexible and solvent-exposed within the native, ligand-bound and amyloidogenic states of muscle acylphosphatase (AcP), a protein previously shown to form amyloid fibrils in the presence of trifluoroethanol. Seven proteases with different degrees of specificity have been used for this purpose. Following exposure to the aggregating conditions, a number of sites along the sequence of AcP become susceptible to proteolytic digestion. The pattern of proteolytic cleavages obtained under these conditions is considerably different from that of the native and ligand-bound conformations and includes a portion within the N-terminal tail of the protein (residues 6-7), the region of the sequence 18-23 and the position 94 near the C terminus. There is a significant overlap between the regions of the sequence found to be solvent-exposed from the present study and those previously identified to be critical in the rate-determining steps of aggregation from protein engineering approaches. This indicates that a considerable degree of solvent exposure is a feature of the portions of a protein that initiate the process of aggregation. PMID- 14741221 TI - Creation of an allosteric enzyme by domain insertion. AB - Two allosteric enzymes have been created by the covalent linkage of non interacting, monomeric proteins with the prerequisite effector-binding and catalytic functionalities, respectively. This was achieved through a combinatorial process called random domain insertion. The fragment of the TEM-1 beta-lactamase gene coding for the mature protein lacking its signal sequence was randomly inserted into the Escherichia coli maltose-binding protein (MBP) gene to create a domain insertion library. This library's diversity derived both from the site of insertion and from a distribution of tandem duplications or deletions of a portion of the MBP gene at the insertion site. From a library of approximately 2 x 10(4) in-frame fusions, approximately 800 library members conferred a phenotype to E.coli cells that was consistent with the presence of bifunctional fusions that could hydrolyze ampicillin and transport maltose in E.coli. Partial screening of this bifunctional sublibrary resulted in the identification of two enzymes in which the presence of maltose modulated the rate of nitrocefin hydrolysis. For one of these enzymes, the presence of maltose increased k(cat) by 70% and k(cat)/K(m) by 80% and resulted in kinetic parameters that were almost identical to TEM-1 beta-lactamase. Such an increase in activity was only observed with maltooligosaccharides whose binding to MBP is known to induce a conformational change. Modulation of the rate of nitrocefin hydrolysis could be detected at maltose concentrations less than 1 microM. Intrinsic protein fluorescence studies were consistent with a conformational change being responsible for the modulation of activity. PMID- 14741222 TI - A chaperone network for the resolubilization of protein aggregates: direct interaction of ClpB and DnaK. AB - The molecular chaperones ClpB (Hsp104) and DnaK (Hsp70) co-operate in the ATP dependent resolubilization of aggregated proteins. A sequential mechanism has been proposed for this reaction; however, the mechanism and the functional interplay between both chaperones remain poorly defined. Here, we show for the first time that complex formation of ClpB and DnaK can be detected by using various types of affinity chromatography methods. The finding that the DnaK chaperone of Escherichia coli is not co-operating with ClpB from Thermus thermophilus further strengthens the specificity of this complex. The affinity of the complex is weak and interaction between both chaperones is nucleotide dependent. The presence of ADP, which is shown to cause dissociation of ClpB(Tth), as well as ClpB deletion mutants incapable of oligomer formation prevent ClpB-DnaK complex formation. The experiments presented indicate a correlation between the oligomeric state of ClpB and its ability to interact with DnaK. The chaperone complex described here might facilitate transfer of intermediates between ClpB and DnaK during refolding of substrates from aggregates. PMID- 14741223 TI - Association and dissociation kinetics for CheY interacting with the P2 domain of CheA. AB - The chemotaxis system of Escherichia coli makes use of an extended two-component sensory response pathway in which CheA, an autophosphorylating protein histidine kinase (PHK) rapidly passes its phosphoryl group to CheY, a phospho-accepting response regulator protein (RR). The CheA-->CheY phospho-transfer reaction is 100 1000 times faster than the His-->Asp phospho-relays that operate in other (non chemotaxis) two-component regulatory systems, suggesting that CheA and CheY have unique features that enhance His-->Asp phospho-transfer kinetics. One such feature could be the P2 domain of CheA. P2 encompasses a binding site for CheY, but an analogous RR-binding domain is not found in other PHKs. In previous work, we removed P2 from CheA, and this decreased the catalytic efficiency of CheA- >CheY phospho-transfer by a factor of 50-100. Here we examined the kinetics of the binding interactions between CheY and P2. The rapid association reaction (k(assn) approximately 10(8)M(-1)s(-1) at 25 degrees C and micro=0.03 M) exhibited a simple first-order dependence on P2 concentration and appeared to be largely diffusion-limited. Ionic strength (micro) had a moderate effect on k(assn) in a manner predictable based on the calculated electrostatic interaction energy of the protein binding surfaces and the expected Debye-Huckel shielding. The speed of binding reflects, in part, electrostatic interactions, but there is also an important contribution from the inherent plasticity of the complex and the resulting flexibility that this allows during the process of complex formation. Our results support the idea that the P2 domain of CheA contributes to the overall speed of phospho-transfer by promoting rapid association between CheY and CheA. However, this alone does not account for the ability of the chemotaxis system to operate much more rapidly than other two-component systems: k(cat) differences indicate that CheA and CheY also achieve the chemical events of phospho-transfer more rapidly than do PHK-RR pairs of slower systems. PMID- 14741225 TI - Additive effects of leptin and topiramate in reducing fat deposition in lean and obese ob/ob mice. AB - The objective of the present study was to investigate the effects of the antiepileptic drug topiramate (TPM) on components of energy balance in lean and obese (ob/ob) mice in the presence or absence of leptin. Lean and ob/ob mice infused with either leptin or phosphate-buffered saline were treated with TPM for 7 days. TPM was mixed into the diet and administered at a dose of 60 mg/kg/day, whereas leptin was infused at the rate of 100 microg/kg/day using osmotic minipumps, which were subcutaneously implanted in the interscapular region. Food intake and body weight were monitored throughout the study. Body composition was measured prior to and following treatment with TPM and leptin, using dual-energy X-ray absorptiometry (DEXA). Glucose (glucose oxidase method) and insulin (radioimmunoassay) were also determined. TPM and leptin significantly reduced body weight gain, food intake and body fat gain in obese mice. The effects of TPM and leptin on fat gain were also statistically significant in lean animals. There was no interaction of TPM and leptin on the energy balance variables, the effects of the two substances being additive instead. Leptin abrogated hyperinsulinemia in obese mutants whereas TPM did not alter insulin levels in either lean or obese mice. The combination of leptin and TPM led to the normalization of glucose levels in obese mice. Our study demonstrates an effect of TPM in leptin-deficient animals, which suggests that TPM does not require the presence of leptin to exert its effect. They also show that the effects of leptin and TPM can be additive. The treatment with leptin in ob/ob mice neither accentuated nor blunted the effect of TPM on energy balance. PMID- 14741226 TI - The distinctiveness of ionic and nonionic bitter stimuli. AB - The diverse chemical structures of stimuli that are bitter to humans suggest a need for multiple bitter receptors. Reactions of golden hamsters (Mesocricetus auratus) to 1 mM quinine hydrochloride, 3 mM denatonium benzoate, 180 mM magnesium sulfate, 30-100 mM caffeine, and 1-1.5 mM sucrose octaacetate (SOA) were studied to address whether there are multiple sensations elicited by bitter stimuli. Methods included behavioral generalization of LiCl-induced conditioned taste aversions (CTAs), intake preference tests, and electrophysiological recordings from the chorda tympani (CT) nerve. The five compounds, all bitter to humans, were all innately aversive to hamsters. CTA for the ionic quinine.HCl, denatonium benzoate, and MgSO(4) mutually cross-generalized and these ionic compounds were effective CT stimuli. Yet, the hamsters were much less sensitive to denatonium than humans, requiring a 100,000 times higher concentration for detection. CTA for nonionic caffeine and SOA did not cross-generalize to quinine or the other two ionic stimuli and these nonionic compounds were not effective CT stimuli. SOA and caffeine may elicit aversive reflexes or systemic reactions rather than taste sensations in the animals. Thus, the three ionic and two nonionic compounds form separate aversive stimulus classes in hamsters, neither of which appears to be a close homologue of the human bitter taste. PMID- 14741228 TI - Nyctohemeral differences in response to restraint stress in CD-1 and C57BL/6 mice. AB - Restraint represents psychological and physical stress. Methods used to model restraint stress in mice vary in duration, time of day during which restraint is applied, and the strain of mouse tested. The goals of this study were: (1) to identify the optimal daily time periods during which the stress response is maximized, and (2) to describe mouse strain differences, if any, in response to restraint. Groups of outbred CD-1 and inbred C57BL/6 mice were restrained for 3 h during three time points of the daily light-dark cycle: (1) the late light phase, (2) the transition between the light phase and the dark phase, and (3) the mid dark phase. Additional mice served as control groups for food deprivation or were unhandled except for blood sampling. Mice of both strains lost significant body mass after 3 days of restraint. Unrestrained food-deprived mice lost body mass, particularly if food-deprived during transition periods. Corticosterone was elevated in restrained mice compared with control mice. Neither basal nor postrestraint corticosterone differed between strains. Corticosterone was elevated by food deprivation during transitional periods in CD-1 mice and during both transition and dark phases in C57 mice. Corticosterone response in restrained CD-1 mice was increased during the dark phase. These results suggest that the physiological response to restraint is similar in both strains. However, corticosterone responses to both restraint and food deprivation were highest during the transitional and dark phases. PMID- 14741227 TI - Effects of intrastriatal administration of selective dopaminergic ligands on spontaneous stereotypy in mice. AB - Abnormal repetitive behaviors are often associated with specific developmental, genetic, and neuropsychiatric disorders. Repetitive motor behaviors, often referred to as stereotypies, have been studied extensively as they can be readily induced by certain drugs (e.g., amphetamine). Recent work has shown, however, that such drug-induced models of stereotypy may not accurately reflect the neurobiological perturbations responsible for the spontaneous manifestation of these behaviors. The present study employed the deer mouse model of spontaneous and persistent stereotypy to evaluate the capacity of several selective dopaminergic agonists (apomorphine, SKF81297, and quinpirole) to exacerbate levels of spontaneously emitted stereotypic topographies when administered intrastriatally. Additionally, the effects of intrastriatal administration of the D(2)R antagonist raclopride on the expression of spontaneous stereotypic jumping were evaluated. No induction or exacerbation of stereotypy was observed following administration of the selective D(1)- or D(2)- receptor agonists, and the mixed agonist apomorphine induced hyperlocomotion and excessive grooming but failed to exacerbate spontaneous stereotypy. Thus, a dissociation was observed between spontaneously emitted and drug-induced stereotypy, suggesting significant limitations to the use of dopamine agonist-induced stereotypy as a model of clinical stereotyped movement disorder. Furthermore, an unexpected and statistically significant (P<.05) potentiation of locomotor activity was observed following intrastriatal raclopride administration, suggesting major alterations to the modulatory characteristics of the striatal dopaminergic system in these spontaneously stereotypic animals. PMID- 14741229 TI - Validation of a cardiac monitor for measuring heart rate variability in adult female pigs: accuracy, artefacts and editing. AB - Autonomic regulation of cardiac activity during stress has not been clearly defined in farm animals. In part, this is due to the limited availability of affordable ambulatory cardiac monitors capable of accurately monitoring and storing large amounts of data that meet the criteria necessary for heart rate variability analysis. Our objectives were to measure the accuracy of a 24-h Polar RR monitor using gold standard ECG, to examine and categorise any occurring anomalies and to ascertain their impact on the outcome of heart rate variability analysis. Five 1-year-old female pigs (gilts) were socially isolated from their pen mates and cardiac activity was simultaneously measured using two systems, a 24-h Polar RR Recorder and a Telemetric ECG system. The Polar data were manually assessed both against and in isolation of the ECG data to identify anomalous beats, which were then assigned to one of five identified error categories. The anomalies in the Polar data were corrected and statistical comparisons were performed among the three data sets to evaluate the effects of anomalies on heart rate variability analysis. Bland-Altman analysis was used to measure the level of agreement among the ECG, Uncorrected Polar and Corrected Polar data. No anomalies or ectopies were found in the ECG data but 46 anomalies (0.81% of total interbeat intervals [IBI]) were found in the Polar Uncorrected data. Manual identification and editing procedures reduced this error to 0.018%. Most mean heart rate and IBI parameters were unaffected by error (P>.05). Standard deviation (S.D.) and root mean square of successive differences (RMSSD) were 45% and 50% higher when anomalies were present in the data. Artefacts affected the magnitude of the frequency domain indices and overestimated total and parasympathetic activity and underestimated sympathetic activity. The mean difference between ECG and Uncorrected Polar data was 1.36 ms (limits of agreement -69.03 to 71.74 ms). This was greatly improved to 0.36 ms (limits of agreement -5.37 to 6.10 ms) after editing. Overall, even a small proportion of error biased the outcome of heart rate variability analysis. This bias was greatly reduced by correcting the anomalous beats. Bland-Altman analysis demonstrated that when there was error present in the Polar data, it could not be used interchangeably with the ECG data. However, if there were no anomalies present in the data or if they were classified and corrected using the approach in this study, then the two systems could be used interchangeably. PMID- 14741230 TI - Potentiation of panic-like behaviors of the rat by subconvulsive doses of strychnine. AB - The present study was carried out to determine possible panicogenic effects of strychnine administered in subconvulsive doses to rats. Two experiments were conducted to assess two major features of panic in animal models: panic-related flight (through the observation of wild running [WR]) and defensive fights. In the first one, 20 adult male Wistar rats were injected with six different doses of strychnine ranging from 0.5 to 4.0 mg/kg. After 15 min of free observation, the animals were submitted to high-intensity acoustic stimulation and the incidence of WR was recorded. Higher doses of strychnine (above 2.5 mg/kg) easily evoked seizures, but lower doses raised the incidence of WR in a dose-dependent manner. The most effective dose for WR (1.5 mg/kg) was used in the second experiment, in which we investigated the effects of strychnine on sleep deprivation-induced fights (SDIFs) that have defensive characteristics. For this purpose, 40 subjects were submitted to 5 days of REM-sleep deprivation by the single-platform method and were then assigned into two groups, i.e., strychnine vs. control. After the injections, the animals were observed in social groupings for SDIF recordings over a period of 60 min. The strychnine-treated groups had more SDIF than the control groups (P<.05, Mann-Whitney U test). We conclude that the high level of neural excitability promoted by partial blockade of the glycinergic system can contribute to the manifestation of panic reactions. PMID- 14741231 TI - Emotional reactivity in mice may not be inherited but influenced by parents. AB - Heredity is often assimilated to genetic transmission of traits. However, some traits may be socially inherited. This has been described for maternal behaviour as well as for emotional reactivity in rodents such as rats or mice. The aim of the present study was to investigate further this idea using two backcrosses between CB6 or B6C females and C57BL/6 males. Indeed, the experimental groups are genetically identical but may be exposed to very different mothering types. When adults, the offspring were subjected to rodent emotional reactivity tests such as elevated plus maze and free exploration paradigm. Results show that CB6xB6 males exhibit higher emotional reactivity than B6CxB6 and B6 males in all behavioural situations, but these effects are not seen in females. Contrarily to their offspring that show different reactivity even if sharing the same genetical background, CB6 and B6C females display quite identical emotional reactivity. A possible explanation is that emotional reactivity is induced by maternal behaviour rather than transmitted by the parents. PMID- 14741232 TI - Training in inhibitory avoidance causes a reduction of Na+,K+-ATPase activity in rat hippocampus. AB - Compelling evidence has indicated the involvement of Na(+),K(+)-ATPase in the mechanisms of synaptic plasticity. In the present study, we investigated the effect of inhibitory avoidance training on Na(+),K(+)-ATPase activity, at different times after training, in the rat hippocampus. Male adult Wistar rats were trained in a step-down inhibitory avoidance task and compared to those submitted to isolated footshock (0.4 mA) or placed directly onto the platform. Na(+),K(+)-ATPase activity decreased, by 60%, in hippocampus of rats sacrificed immediately after the isolated footshock, as well as immediately (0 min) and 6 h after training; this effect was not present 24 h after training. We also verified that enzyme activity was not altered in rats killed after just being on the platform. These findings suggest that Na(+),K(+)-ATPase activity may be involved in the memory consolidation of step-down inhibitory avoidance in the hippocampus. PMID- 14741233 TI - Attenuation of stress-induced elevations in plasma ACTH level and body temperature in rats by green odor. AB - To investigate the effect of green odor on the elevation of the plasma adrenocorticotropic hormone (ACTH) levels and body temperature (T(b)) induced by stress, adult male rats were subjected to a 2-h immobilization stress and exposed to green odor or its vehicle only. In comparison with the vehicle group, animals in the green odor group showed a significant reduction in plasma ACTH levels at the end of the stress when green odor was applied during the stress. The elevated plasma ACTH levels 2 days after the stress were reduced by green odor applied 0, 1, 2, 4 or 6 h after the beginning of the stress. Neither the immediate nor the long-lasting plasma ACTH response was affected by a prestress treatment of green odor. T(b) elevation was evident following the end of the stress and during the light phase the day after the stress. Both the immediate and the long-lasting elevations in T(b) were attenuated by green odor. These data suggest that green odor extracted from green leaves has a relieving effect on plasma ACTH and T(b) levels induced by an immobilization stress for not only immediate but also long lasting periods. PMID- 14741234 TI - Inhibition of glucose oxidation by alpha-cyano-4-hydroxycinnamic acid stimulates feeding in rats. AB - Alpha-cyano-4-hydroxycinnamic acid (4-CIN, 100-200 mg/kg b.wt.), which impairs glucose oxidation by inhibiting pyruvate transport across the mitochondrial membrane, stimulated feeding in rats following intraperitoneal injection without affecting blood glucose level. Like 2-deoxy-D-glucose (2-DG), an inhibitor of glycolysis, 4-CIN probably acts mainly on the CNS through activation of alpha(2) adrenergic receptors, because the feeding response to 4-CIN was eliminated by phentolamine or yohimbine. Unlike feeding elicited by 2-DG, 4-CIN-induced feeding was eliminated by total abdominal (but not hepatic branch) vagotomy. Since peripheral atropinization also blocked 4-CIN-induced feeding, activation of central parasympathetic neurons seems to be involved in 4-CIN-induced feeding. The feeding response to 4-CIN was diminished in rats fed a high-fat diet, probably because metabolic sensors sensing fatty acid oxidation counteract the feeding response to 4-CIN. The results suggest that inhibition of glucose oxidation by blocking pyruvate entry into mitochondria stimulates feeding in rats in particular when fed a high-carbohydrate diet. PMID- 14741235 TI - Artificial neural network analysis of gustatory responses in the thalamic taste relay of the rat. AB - We used an artificial neural network (ANN) as a model for analyzing single-neuron responses from the thalamic taste relay of rats. The network consisted of: (1) a layer of 44 input units, representing the responses of the 44 thalamic taste cells; (2) a layer of hidden units of varying numbers; and (3) a layer of four output units. We used the back-propagation algorithm to train the output units to discriminate among tastants based on inputs from the thalamic neurons. As the network became fully trained, we found that: (1) only two hidden units were necessary to provide nearly the full discriminative capacity of the network; (2) the loss of even a few of the input units that had the highest impact on hidden units caused a drastic reduction of discriminative power, implying that not all neurons contribute equally to the neural code; and (3) adding a temporal component to the input, by representing each 100-ms time bin as a separate input unit, increased the accuracy with which output units were able to identify tastants. We used data from behavioral discrimination tasks as a measure of the capacity of the network to identify stimuli correctly. A network with two hidden units was about as effective as an across-pattern analysis in accounting for the rat's discriminative ability. PMID- 14741236 TI - Correlation between driving errors and vigilance level: influence of the driver's age. AB - During long and monotonous driving at night, most drivers progressively show signs of visual fatigue and loss of vigilance. Their capacity to maintain adequate driving performance usually is affected and varies with the age of the driver. The main question is to know, on one hand, if occurrence of fatigue and drowsiness is accompanied by a modification in the driving performance of the driver and, on the other hand, if this relationship partially depends on the driver's age. Forty-six male drivers, divided into three age categories: 20-30, 40-50, and 60-70 years, performed a 350-km motorway driving session at night on a driving simulator. Driving errors were measured in terms of number of running-off the-road incidents (RORI) and large speed deviations. The evolution of physiological vigilance level was evaluated using electroencephalography (EEG) recording. In older drivers, in comparison with young and middle-aged drivers, the degradation of driving performance was correlated to the evolution of lower frequency waking EEG (i.e., theta). Contrary to young and middle-aged drivers, the deterioration of the vigilance level attested by EEG correlated with the increase in gravity of all studied driving errors in older drivers. Thus, depending on the age category considered, only part of the driving errors would constitute a relevant indication as for the occurrence of a state of low arousal. PMID- 14741237 TI - Development of defensive behavior and conditioning to cat odor in the rat. AB - Laboratory rats show a range of defensive behaviors, including freezing, avoidance, and risk assessment upon exposure to cat odor, an unconditioned but highly effective threat stimulus. This study examined defensive behaviors, and the rapid conditioning to context plus cue, of these behaviors, in 18-, 26-, and 38-day-old male and female rats exposed to cat odor. Rats were placed individually in a runway with a cloth covered (control or saturated with cat fur/skin odor) block for a 10-min trial. On the following day, a similar trial involved an odorless block. On the odor exposure day, rats of all ages showed less contact with the odor block than with the control block. The 26- and 38-day old rats, but not the 18-day-old rats, also showed locomotor suppression, more avoidance of the area where the odor block was located, and more risk assessment than no-odor controls. On a test of conditioned behavior 24 h following exposure, 26- and 38-day-old rats exhibited defensive behavior including avoidance and reduction of locomotion while 18-day-old pups did not. PMID- 14741238 TI - Facilitation of aggressive and sexual behaviors by saccharin deprivation in rats. AB - The 'deprivation effect' (DE) phenomenon is expressed as an increase in the level of free choice consumption of drugs, alcohol, or saccharin following a period of forced abstinence in humans and in several species of laboratory animals. The DE may reflect relapse-like drinking and be relevant for modeling addictive behaviors. In humans, drug or alcohol abstinence is commonly associated with the increased physical and sexual abuse. The present study aimed to study whether aggressive and sexual behaviors are affected by the conditions known to result in the DE. Adult male Wistar rats had unlimited free choice access to water and saccharin-containing solution (0.1%, w/v). After the preference for saccharin was established, animals underwent repeated 7-day-long episodes of saccharin deprivation. It was found that (i) aggressive and sexual behaviors were facilitated after 7 days of saccharin deprivation, and (ii) DE was significantly reduced in animals that were allowed to interact with the nonaggressive male or receptive female conspecifics during the last day of deprivation exposure. These results suggest that the saccharin deprivation exposures may facilitate aggressive and sexual behaviors. Alternative behavioral strategy may serve as a mechanism of coping with the deprivation state. PMID- 14741239 TI - Effect of response to backtest and housing condition on cell-mediated and humoral immunity in adult pigs. AB - Several recent studies in juvenile pigs demonstrated a relationship between the degree of resistance displayed early in life in a so-called "backtest" and parameters of cell-mediated and humoral immunity. Some of the immune characteristics were reported to depend on the interaction between backtest classification and housing system. In the present study, the effects of backtest classification and housing condition on immune reactivity in adult gilts were examined. At 10 and 17 days of age, female piglets were subjected to the backtest. In this test, each piglet is restrained on its back for 1 min and the number of escape attempts is scored. Pigs classified as high resisting (HR) or low resisting (LR) were selected and housed in groups of six gilts. At 7 months of age, half of the gilts were housed in individual stalls. At 12 months of age, gilts were challenged by immunization with DNP-KLH. Control gilts were treated similarly with a placebo. Blood samples were drawn prior to immunization (Day 0) and weekly thereafter until Day 28. No significant effects of backtest type on cellular and humoral responses against KLH were found. Furthermore, being housed in stalls as compared to groups had no consequences for the immune response and did not induce differences between HR and LR gilts. Differences in behavior and physiology found previously between HR and LR gilts, particularly in gilts in stall housing, may thus be of relatively little importance for immune-related health. PMID- 14741240 TI - Dominance status predicts response to nonsocial forced movement stress in the green anole lizard (Anolis carolinensis). AB - We used changes in body color and eyespot formation, two somatic indices of stress controlled mainly by catecholamine activity, to compare the reactions of dominant and subordinate male green anole lizards (Anolis carolinensis) to a nonsocial stressor, forced movement. Individual males were pretested by subjecting them to 10 min of forced movement induced by chasing them around their home cage with a slender wooden stick. Stress responses were assayed via changes in body color (progressive darkening from green to brown indicating increasing stress) and expression of a black postorbital eyespot (which appears with increasing catecholaminergic stress responses). Lizards were paired and allowed to form stable dominant/subordinate relationships for 2 weeks. After that period of stable social status, dominants and subordinates were separated and subjected to the same forced-movement stress. There was no difference between experimental groups in the pretest. After assuming positions in the dominance hierarchy, however, dominant males showed reduced somatic indicators of stress and were quicker to recover from the stress. The data suggest that animals that assumed the dominant position decreased their stress response relative to the pretest, while animals that assumed the subordinate position increased their stress response relative to the pretest. The results indicate that dominant social status may have advantages beyond the realm of social interactions by enhancing an individual's ability to tolerate other, nonsocial stressful events. PMID- 14741241 TI - Taste modulation of nociception differently affects chronically stressed rats. AB - Stress responses cover a wide range of physiological changes, including alterations in the perception of and response to pain. Animals submitted to repeated stress present altered nociception and this effect is part of this process of adaptation; in addition pleasant and unpleasant experiences with tastes and odors have been shown to affect distinct behavioral aspects, such as pain perception. The aim of the present study is to verify the responses of repeatedly stressed rats (1 h of daily immobilization during 40 days) to pleasant and unpleasant tastes on nociception, when compared to control animals. An increase in the tail-flick latency (TFL) was observed 5 min after exposure to a sweet taste in the control group, whereas no effect was observed in chronically stressed animals. When submitted to an unpleasant taste (5% acetic acid), the chronically stressed group presented an increase in TFL, whereas no effect was observed in the control group. In conclusion, chronically stressed animals present different nociceptive responses to sweet and acid tastes; although control animals suitably respond to a sweet stimulus, stressed animals seem to be more apt to react to the unpleasant stimulus. PMID- 14741242 TI - Systemic nicotine alters whole-body fat utilization in female rats. AB - Cigarette smoking produces weight suppression in humans that often rebounds following smoking cessation. Nicotine (NIC) administration produces similar effects in rats. While changes in food intake are thought to account for some of the body weight changes, few reports have investigated how NIC affects whole-body metabolism. In the present study, measures of respiratory quotient (RQ) and energy expenditure (EE) were used to investigate metabolic changes that may contribute to NIC's effects on body weight. Female rats (n=46) were implanted for 14 days with subcutaneous Alzet minipumps containing NIC (6 mg/kg/day) or its vehicle. One-hour metabolic test sessions occurred four times: 2 and 12 days after pump implant and 2 and 8 days after pump removal. NIC initially suppressed body weight gain, followed by steady recovery that was briefly exaggerated after withdrawing NIC. Daily food intake was acutely suppressed by NIC and acutely potentiated after NIC cessation. RQ, but not EE, was suppressed by NIC 2 days after pump implant indicating increased fat utilization. Conversely, RQ was increased 2 days after pump removal signaling increased fat storage. These findings indicate that acute changes in whole-body metabolism may contribute to the weight modulating effects of NIC. PMID- 14741243 TI - Stress-induced modification of anxiety in rats is dependent on reproductive status. AB - Pregnancy and lactation are accompanied by behavioral changes that include altered responses to stress. Previous exposure to a stressor can also modulate subsequent stress-related behavior, and this effect is at least partly mediated by the reproductive hormone, estrogen. In this study, we sought to determine if the stress-related behavior of rats that had been exposed to a cat would vary with reproductive status. After exposure to a cat or a nonstress control procedure, pregnant, lactating, virgin female or male rats received two acoustic startle test sessions on consecutive days, followed 4 days later by testing on an elevated plus maze. Startle amplitudes tended to increase across trials in all groups, but were differentially enhanced in nonstressed pregnant and lactating rats. A 5-min exposure to a cat eliminated the enhanced responding in these two groups. The cat exposure attenuated startle stimulus-evoked freezing only in the pregnant rats and produced differential plus maze performance in this group. Cat exposure produced no differential effects in virgin females or males. These results suggest that reproductive state can influence the impact of a stressor on subsequent behavior, but does so in a rather complex way. PMID- 14741244 TI - Effects of acute exercise on the soleus H-reflex and self-reported anxiety after caffeine ingestion. AB - The effects of moderate intensity cycling exercise on the soleus H-reflex and state anxiety were examined among 16 individuals whose anxiety was experimentally manipulated by consumption of a large dose of caffeine. The soleus H-reflex and state anxiety were measured before and 1 h after consuming caffeine or placebo and then again 10 min after 30 min of either cycling at an intensity of 60% VO(2peak) or quiet rest. We found that (1) caffeine consumption did not influence the amplitude of the soleus H-reflex, but it did increase state anxiety; (2) acute exercise reduced the soleus H-reflex after consumption of either caffeine or placebo, but it reduced state anxiety only after consumption of caffeine; and (3) there was no evidence of a relationship between changes in the soleus H reflex and state anxiety. Exercise-induced anxiolysis does not appear to underlie the postexercise reduction of the soleus H-reflex. PMID- 14741245 TI - Inhibition kinetics of carba- and C-fucosyl analogues of GDP-fucose against fucosyltransferase V: implication for the reaction mechanism. AB - Inhibition kinetics of two isosteric analogues of GDP-fucose (GDP-Fuc) were investigated against fucosyltransferase V using electrospray ionization mass spectrometry coupled to multiple reaction monitoring. The carba-Fuc analogue was found to be a competitive inhibitor with a K(i) value of 67.1+/-9.8 microM, similar to the K(m) value for GDP-Fuc (50.4+/-5.5 microM), while the C-Fuc analogue exhibited significantly weak competitive inhibition with a K(i) value of 889+/-93 microM. PMID- 14741246 TI - Synthesis and biological evaluation of (-)-laulimalide analogues. AB - Analogues of the marine natural product (-)-laulimalide were prepared by total synthesis and evaluated in vitro for anticancer activity. PMID- 14741247 TI - Synthesis and incorporation of an alpha-hexofuranosyl thymidine into oligodeoxynucleotides via its two exocyclic OH-groups. AB - 1-(2,3-Dideoxy-3-amino-alpha-D-arabino-hexofuranosyl)thymine is considered as a conformationally restricted acyclic nucleoside using the furanose ring to link the diol backbone to the nucleobase. The appropriately substituted phosphoramidites were synthesised via 1-(5,6-di-O-acetyl-2,3-dideoxy-3 phthalimido-alpha-D-arabino-hexofuranosyl)thymine and used in oligodeoxynucleotide (ODN) synthesis. However, the binding affinity of the mixed ODNs towards complementary DNA and RNA was decreased compared to the wild-type oligos. The decrease was smaller when the monomer was inserted near the end of the sequence. The insertions into an alpha T sequence or in a beta T sequence gave nearly the same dropping in melting temperature per modification which indicates that the new nucleotide modifications behave both as alpha and beta nucleotides. PMID- 14741248 TI - Chemoenzymatic synthesis and binding affinity of novel (R)- and (S)-3-aminomethyl 1-tetralones, potential atypical antipsychotics. AB - A series of (R)- and (S)-3-aminomethyl-1-tetralones, conformationally constrained analogues of haloperidol, have been obtained by enzymatic resolution of the corresponding racemic 3-hydroxymethyl-1-tetralones using Pseudomonas fluorescens lipase. Their binding affinities at dopamine D(2) and serotonin 5-HT(2A) and 5 HT(2C) receptors were determined showing in some cases an atypical antipsychotic profile with Meltzer's ratio higher than 1.30. PMID- 14741250 TI - Synthesis of new thiophene and benzo[b]thiophene hydrazide derivatives as human NPY Y(5) antagonists. AB - Neuropeptide Y is one of the most potent appetite stimulating hormones known. Novel thiophene and benzo[b]thiophene hydrazide derivatives were synthetized and evaluated biologically as NPY Y(1) and Y(5) receptor subtype antagonists. They were found to have nanomolar binding affinities for human NPY Y(5) receptor, obtaining the lead compound, trans-N-4-[N'-(thiophene-2 carbonyl)hydrazinocarbonyl]cyclohexylmethyl-4-bromobenzenesulfonamide, which binds with a 7.70 nM IC(50) to the hY(5) receptor. PMID- 14741249 TI - Aza-bicyclic amino acid sulfonamides as alpha(4)beta(1)/alpha(4)beta(7) integrin antagonists. AB - The design, synthesis, and biological activity of novel alpha(4)beta(1) and alpha(4)beta(7) integrin antagonists, containing a bridged azabicyclic nucleus, are reported. Conformational analysis of targets containing an azabicyclo[2.2.2]octane carboxylic acid and known integrin antagonists indicated that this azabicycle would be a suitable molecular scaffold. Variation of substituents on the pendant arylsulfonamide and phenylalanine groups resulted in potent alpha(4)beta(1)-selective and dual alpha(4)beta(1)/alpha(4)beta(7) antagonists. Potent compounds 11i, 11h, and 14 were effective in the antigen sensitized sheep model of asthma. PMID- 14741251 TI - Rational design and synthesis of selective BACE-1 inhibitors. AB - An effective approach for enhancing the selectivity of beta-site amyloid precursor protein cleaving enzyme (BACE 1) inhibitors is developed based on the unique features of the S1' pocket of the enzyme. A series of low molecular weight (<600) compounds were synthesized with different moieties at the P1' position. The selectivity of BACE 1 inhibitors versus cathepsin D and renin was enhanced 120-fold by replacing the hydrophobic propyl group with a hydrophilic propionic acid group. PMID- 14741252 TI - Inhibition of estrone sulfatase (ES) by alkyl and cycloalkyl ester derivatives of 4-[(aminosulfonyl)oxy] benzoic acid. AB - In our search for potent inhibitors of the enzyme estrone sulfatase (ES), we have undertaken the synthesis and biochemical evaluation of a range of esters of 4 [(aminosulfonyl)oxy] benzoic acid. The results of the study show that the synthesised compounds possess potent inhibitory activity, indeed the cyclooctyl derivative was found to be more potent than 667-COUMATE, which is currently undergoing clinical trials. PMID- 14741253 TI - Synthesis and biological evaluation of novel coumarin derivatives with a 7 azomethine linkage. AB - The synthesis of several novel coumarin derivatives with a 7-azomethine linkage was carried out starting from 7-formyl-coumarin. The compounds were tested in vivo for their anti-inflammatory activity and in vitro for their antioxidant ability. Compounds 3a and 3e possess significant protection against carrageenin induced rat paw edema. PMID- 14741254 TI - Diamine derivatives with antiparasitic activities. AB - There are a lack of effective chemotherapies for many parasitic diseases. Polyamine pathways have been reported as potential targets for the development of new chemotherapies against parasitic diseases. In the present study, different libraries of substituted diamines totalling 78 compounds have been synthesized on solid support and their activities against malaria and leishmania parasites have been determined. Most active compounds demonstrated submicromolar activities against both organisms and their structure-activity relationships are discussed. PMID- 14741255 TI - Synthesis and antitumor activity of 10-substituted benzylidene anthrone. AB - Fifteen compounds of 10-substituted benzylidene anthrone were prepared with moderate yield by reaction of anthrone and substituted benzaldehydes under the presence of pyridine and piperidine as catalyst. Their antitumor activities in vitro were evaluated. The results show that the electron-withdrawing substitutes decrease the activities, the electron-donor substitutes increase the activities; the compound with substitute at ortho or para position has stronger activities than that of compound with the same substitute but located at the meta position. There are six compounds which appear as strong effective inhibition for A-549 cancer cell growth. This is a kind of good leading compound which is worth researching further. PMID- 14741256 TI - Permeation and metabolism of a series of novel lipophilic ascorbic acid derivatives, 6-O-acyl-2-O-alpha-D-glucopyranosyl-L-ascorbic acids with a branched acyl chain, in a human living skin equivalent model. AB - A series of novel lipophilic vitamin C derivatives, 6-O-acyl-2-O-alpha-D glucopyranosyl-L-ascorbic acids possessing a branched-acyl chain of varying length from C(8) to C(16) (6-bAcyl-AA-2G), were evaluated as topical prodrugs of ascorbic acid (AA) with transdermal activity in a human living skin equivalent model. The permeability of 6-bAcyl-AA-2G was compared with those of the derivatives having a straight-acyl chain (6-sAcyl-AA-2G). Out of 10 derivatives of 6-sAcyl-AA-2G and 6-bAcyl-AA-2G, 6-sDode-AA-2G and 6-bDode-AA-2G exhibited most excellent permeability in this model. Measurement of the metabolites permeated from the skin model suggested that 6-bDode-AA-2G was mainly hydrolyzed via 6-O-acyl AA to AA by tissue enzymes, while 6-sDode-AA-2G was hydrolyzed via 2 O-alpha-D-glucopyranosyl-L-ascorbic acid to AA. The former metabolic pathway seems to be advantageous for a readily available source of AA, because 6-O-acyl AA, as well as AA, is able to show vitamin C activity. PMID- 14741258 TI - Synthesis and biological activity of a novel squalene epoxidase inhibitor, FR194738. AB - The synthesis and biological properties of a novel squalene epoxidase inhibitor, FR194738, are described. This compound displayed potent in vitro inhibitory activities against squalene epoxidase and cholesterol synthesis, and lowered plasma cholesterol and triglyceride levels in dogs. PMID- 14741257 TI - Synthesis and mitochondrial complex I inhibition of dihydroxy-cohibin A, non-THF annonaceous acetogenin analogue. AB - To elucidate the inhibitory action of acetogenins, we synthesized an acetogenin derivative which possesses tetraol in place of the tetrahydrofuran ring and examined its inhibitory activity against bovine heart mitochondrial complex I. Our results indicate that these hydroxy groups are an essential structural factor though it is not effective as bis-THF hydroxy groups combination. PMID- 14741259 TI - Macrocyclic piperazinones as potent dual inhibitors of farnesyltransferase and geranylgeranyltransferase-I. AB - A series of macrocyclic piperazinone compounds with dual farnesyltransferase/geranylgeranyltransferase-I inhibitory activity was prepared. These compounds were found to be potent inhibitors of protein prenylation in cell culture. A hypothesis for the binding mode of compound 3o in FPTase is proposed. PMID- 14741260 TI - Degradation of target protein in living cells by small-molecule proteolysis inducer. AB - Ubiquitin-dependent proteolysis of cellular proteins is one of the major pathways to regulate protein function posttranslationally. Here we demonstrate a potentially general method of degrading any targeted proteins by the ubiquitin dependent proteolysis in living cells, using small-molecule proteolysis inducer (SMPI). PMID- 14741261 TI - Biosynthesis of agglomerin A: stereospecific incorporation of pro-R- and pro-S hydrogens at sn-C-3 of glycerol into the branched C(3) moiety. AB - The biosynthetic origin of the C(3) branched unit of agglomerin A has been investigated. Feeding of sn-(3R)- and sn-(3S)-[3-(2)H]glycerols to Enterobacter agglomerans PB-6042 followed by (2)H NMR analysis of the resulting agglomerin A revealed that pro-R and pro-S hydrogens at sn-C-3 of glycerol were incorporated stereospecifically into 5E and 5Z hydrogens of agglomerin A, respectively. These results imply that the immediate precursor of the C(3) branched unit is not pyruvate, but 1,3-bisphosphoglyceric acid or its biological equivalent. PMID- 14741262 TI - Synthesis and antifungal activities of phenylenedithioureas. AB - A total of 20 new phenylenedithiourea derivatives was synthesized by reaction of phenylenediisothiocyanates with aromatic amines as aminobenzoic, aminosalicylic acid and their derivatives. Their chemical structures were confirmed by elemental analysis, IR spectrometry and 1H NMR. The compounds were screened for in vitro antifungal, antibacterial activities and some of them have strong antifungal activities comparable to the activity observed for ketoconazole. PMID- 14741263 TI - Solid-phase synthesis of a small library of 3-phenylthio-3-nicotinyl propionic acid derivatives acting as antagonists of the integrin alphaVbeta3. AB - We describe the synthesis of a series of low molecular weight inhibitors of the alphavbeta3 integrin obtained by modifying a high-throughput screening hit with micromolar activity. A solid phase synthesis to prepare 3-phenylthio-3-nicotinyl propionic acid derivatives, exemplified by 13c, was set up. Compounds with nanomolar activity in the biochemical assay and able to efficiently inhibit cell adhesion mediated by vitronectin have been obtained. PMID- 14741264 TI - TMC-95A, a reversible proteasome inhibitor, induces neurite outgrowth in PC12 cells. AB - TMC-95A has been characterized as a potent proteasome inhibitor that binds to enzymes non-covalently at low nanomolar concentrations. Herein, the neuritogenic activity of TMC-95A in PC12 rat pheochromocytoma cells is reported for the first time. TMC-95A induced a positive neurite initiation of PC12 cells at concentration ranging from 1 to 20 microM. PMID- 14741266 TI - Structure-activity relationships of non-imidazole H(3) receptor ligands. Part 3: 5-Substituted 3-phenyl-1,2,4-oxadiazoles as potent antagonists. AB - Further SAR studies on novel histamine H(3) receptor antagonists are presented. Compound 14bb is a potent antagonist of both the rat cortical and human clone receptors, and is demonstrated to act functionally as an antagonist in an in vivo mouse dipsogenia model. PMID- 14741265 TI - Manipulation of kinetic profiles in 2-aryl propionic acid cyclooxygenase inhibitors. AB - The nonsteroidal anti-inflammatory drugs flurbiprofen and ibuprofen were modified in an attempt to alter the kinetics of inhibitor binding by COX-1. Contrary to prior predictions, a halogen substituent is not sufficient to confer slow tight binding behavior. Conversion of the carboxylate moiety of flurbiprofen to an ester or amide abolishes slow tight-binding behavior, regardless of halogenation state. PMID- 14741267 TI - Thiazoles and thiopyridines: novel series of high affinity h5HT(7) ligands. AB - A series of thiazole based 5HT(7) ligands has been identified from screening. Optimisation of the pendent aryl group and modification of the core gave a related series of high affinity, selective thiopyridine based 5HT(7) ligands, the most active of which behaves as a partial agonist. PMID- 14741268 TI - 2-hydroxy-4-isopropylbenzaldehyde, a potent partial tyrosinase inhibitor. AB - Chamaecin (2-hydroxy-4-isopropylbenzaldehyde) was synthesized and tested for its tyrosinase inhibitory activity. It partially inhibits the oxidation of L-3,4 dihydroxyphenylalanine (L-DOPA) catalyzed by mushroom tyrosinase with an IC(50) of 2.3 microM. The inhibition kinetics analyzed by Dixon plots found that chamaecin is a mixed type inhibitor. This inhibition may come in part from its ability to form a Schiff base with a primary amino group in the enzyme. PMID- 14741269 TI - Peptidyl aldehydes as slow-binding inhibitors of dual-specificity phosphatases. AB - Peptidyl aldehydes were tested for inhibition of dual-specificity phosphatases VH1 and VHR. The most potent compound, cinnamaldehyde-Gly-Glu-Glu (Cinn-GEE), acted as a slow-binding inhibitor with K(I)* values of 18 and 288 microM against VH1 and VHR, respectively. PMID- 14741270 TI - A new class of potent non-imidazole H(3) antagonists: 2-aminoethylbenzofurans. AB - 2-aminoethylbenzofurans constitute a new class of H(3) antagonists that are more rotationally constrained than most previously reported H(3) antagonists. They retain high potency at human and rat receptors, with efficient CNS penetration observed in 35. The SAR of the basic amine moiety was compared in three different series of analogues. The greatest potency was found in analogues bearing a 2 methylpyrrolidine, a 2,5-dimethylpyrrolidine, or a 2,6-dimethylpiperidine. PMID- 14741271 TI - Synthesis and evaluation of novel bacterial rRNA-binding benzimidazoles by mass spectrometry. AB - A series of novel benzimidazoles were efficiently synthesized using both solution and solid-phase chemistry. These compounds were found to bind to the bacterial 16S ribosomal RNA A-site with micromolar affinities using unique mass spectrometry-based assays. PMID- 14741272 TI - Inhibition of alpha-chymotrypsin with thiol-bearing substrate analogues in the presence of zinc ion. AB - We have demonstrated that thiol-bearing analogues of alpha-chymotrysin (alpha-CT) substrates such as (S)-(1-benzyl-2-thiolethyl)-carbamic acid, benzyl ester (3) inhibits alpha-CT, a prototypical serine protease, in the presence of Zn(II) ion. They constitute a novel class of small molecule inhibitors for alpha-CT believed to inhibit the enzyme by forming a ternary complex consisting of alpha-CT, Zn(II) ion, and the inhibitor. PMID- 14741273 TI - QSAR studies of PC-3 cell line inhibition activity of TSA and SAHA-like hydroxamic acids. AB - Quantitative structure-activity relationships (QSAR) for a series of new trichostatin A (TSA)-like hydroxamic acids for the inhibition of cell proliferation of the PC-3 cell line have been developed using molecular descriptors from Qikprop and electronic structure calculations. The best regression model shows that the PM3 atomic charge on the carbonyl carbon in the CONHOH moiety(Qco), globularity (Glob), and the hydrophilic component of the solvent-accessible surface area (FISA) describe the IC(50) of 19 inhibitors of the PC-3 cell line with activities ranging over five orders of magnitude with an R(2)=0.92 and F=59.2. This information will be helpful in the further design of novel anticancer drugs for treatment of prostate cancer and other diseases affected by HDAC inhibition. PMID- 14741274 TI - Rational design of azepane-glycoside antibiotics targeting the bacterial ribosome. AB - RNA recognition by natural aminoglycoside antibiotics depends on the 2 deoxystreptamine (2-DOS) scaffold which participates in specific hydrogen bonds with the ribosomal decoding-site target. Three-dimensional structure information has been used for the design of azepane-monoglycosides, building blocks for novel antibiotics in which 2-DOS is replaced by a heterocyclic scaffold. Azepane glycosides showed target binding and translation inhibition in the low micromolar range and inhibited growth of Staphylococcus aureus, including aminoglycoside resistant strains. PMID- 14741275 TI - Design of small molecule ketoamide-based inhibitors of cathepsin K. AB - A novel series of ketoamide-based inhibitors of cathepsin K has been identified. Modifications to P(2) and P(3) elements were crucial to enhancing inhibitory activity. Although not optimized, a selected inhibitor was effective in attenuating type I collagen hydrolysis in a surrogate assay of bone resorption. PMID- 14741277 TI - 3-(2-pyrrolidin-1-ylethyl)-5-(1,2,3,6-tetrahydropyridin-4-yl)-1H-indole derivatives as high affinity human 5-HT(1B/1D) ligands. AB - A series of 3-(2-pyrrolidin-1-ylethyl)-5-(1,2,3,6-tetrahydropyridin-4-yl)-1H indole derivatives (2) has been prepared using parallel synthesis techniques, and their structure-activity relationships studied. High affinity human 5-HT(1B/1D) (h5-HT(1B/1D)) ligands have been identified. PMID- 14741276 TI - Enantioselective ester hydrolysis catalyzed by beta-cyclodextrin conjugated with beta-hairpin peptides. AB - Designed cyclodextrin-peptide conjugates, which have one or two beta-hairpin peptides, have been synthesized as catalysts for ester hydrolysis. One or two beta-hairpin peptides were located at the primary hydroxyl group side of beta cyclodextrin so as to arrange two histidine residues that act as a general acid and a general base catalysts and provide the substrate recognition subsite. Kinetic studies revealed that the two-beta-hairpin peptide was more effective than that of the one-beta-hairpin peptide for substrate recognition. PMID- 14741278 TI - 3D-QSAR studies on PU3 analogues by comparative molecular field analysis. AB - A comparative molecular field analysis (CoMFA) of PU3 derivatives of Hsp90 (Heat shock protein 90) inhibitors has been performed to determine the factors contributing the corresponding activities. The energy minimized conformations were obtained by molecular mechanics using SYBYL package. The developed model, with r(2) value of 0.947, was verified by performing leave-one out (LOO) cross validation, which showed q(2) value of 0.513. The calculated model not only elucidates the relationship between compound structures and biological activities but, more importantly, facilitates design of new Hsp90 inhibitors with calculated antiproliferative activity. PMID- 14741279 TI - Vancomycin disulfide derivatives as antibacterial agents. AB - A series of lipidated vancomycin analogues 1 bearing disulfide bonds within their lipid chains was designed and synthesized to optimize their ADME profiles while retaining antibacterial potency. These compounds exhibited good activity against resistant organisms and low accumulation in tissues such as kidney and liver. PMID- 14741280 TI - Novel nevirapine-like inhibitors with improved activity against NNRTI-resistant HIV: 8-heteroarylthiomethyldipyridodiazepinone derivatives. AB - A series of 8-heteroarylthiomethyldipyridodiazepinone derivatives were prepared and evaluated for their antiviral profile against wild type virus and the important K103N/Y181C mutant as an indicator for broad activity. 2,6 Dimethylpyridine derivative 16 was found to have a good pharmacokinetic profile in spite of poor metabolic stability in rat liver microsomes. PMID- 14741281 TI - Fluorinated dihydroquinolines as potent n-NOS inhibitors. AB - Fluorinated dihydroquinolines showed reduced basicity of the amidine function. Their syntheses and potencies as neuronal nitric oxide synthase (n-NOS) inhibitors are reported. PMID- 14741282 TI - Synthesis and biological results of the technetium-99m-labeled 4-nitroimidazole for imaging tumor hypoxia. AB - 1-(4-Nitroimidazole-1-yl)-propanhydroxyiminoamide (N4IPA) was synthesized. The biodistribution of 99mTc-N4IPA in mice bearing S180 tumor demonstrated that the complex showed accumulation in tumor and slow clearance from it. The tumor-to tissue uptake ratios increase with time. These results suggest that 99mTc-N4IPA would be a marker for imaging tumor hypoxia. PMID- 14741283 TI - Benzene-di-N-octylcarbamates as conformationally constrained phospholipase A(2) inhibitors. AB - Conformationally constrained 1,2-, 1,3-, and 1,4-benzene-di-N-octylcarbamates are potent reversible competitive inhibitors of Naja mocambique mocambique phospholipase A(2) with the K(i) values of 11, 4, and 15 microM, respectively. With the angle of 120(o) between two C(benzene)-O bonds, 1,3-benzene-di-N octylcarbamate mimics the preferable eclipsed C(sn-2)-O/C(sn-3)-O conformer of phospholipid in the enzyme-phospholipid complex. Further, a three-step phospholipase A(2) inhibition mechanism by the inhibitor is proposed. PMID- 14741284 TI - Synthesis and antibiotic activity of the tricyclic furo[3,2-c] isochromen-2 trione unit of the pyranonaphthoquinones. AB - The elaboration and biological activity of 15, containing the proposed pharmacophore for the antibiotic activity of the pyranonaphthoquinones, are reported. The synthetic strategy involved acid-catalyzed lactonization of mandelate 17 for isochroman ring formation, in combination with a Wittig-oxa Michael functionalization of isochroman-3-ol derivative 20, a lactonization involving configurational inversion of a benzylic alcohol and a final AgO oxidation. Compound 15 showed activity against Staphylococcus aureus and Bacillus subtilis with MIC of 64 and 32 microg/mL, respectively. PMID- 14741285 TI - Synthesis and biological evaluation of piperazine-based derivatives as inhibitors of plasminogen activator inhibitor-1 (PAI-1). AB - Compound 2 was identified by high throughput screening as a novel PAI-1 inhibitor. Systematic optimization of the A, B, and C segments of 2 resulted in the identification of a more potent compound 39 with good oral bioavailability. The synthesis and SAR data are presented in this report. PMID- 14741286 TI - Synthesis of N,N'-disubstituted 3-aminobenzo[c] and [d]azepin-2-ones as potent and specific farnesyl transferase inhibitors. AB - A structure-activity study was performed by synthesis on N,N'-disubstitution of 3 aminobenzo[c] and [d]azepin-2-one 2 and 3 to afford potent and specific farnesyl transferase inhibitors with low nM enzymatic and cellular activities. PMID- 14741287 TI - Bile acid amides derived from chiral amino alcohols: novel antimicrobials and antifungals. AB - Cholic and deoxycholic acid amides 10-17 have been synthesised from (1R,2R)-1 phenyl-2-amino-1,3-propanediol 2, (1S,2S)-1-phenyl-2-amino-1,3-propanediol 4, (1R,2R)-1-para-nitrophenyl-2-amino-1,3-propanediol 3, (1S,2S)-1-para-nitrophenyl 2-amino-1,3-propanediol 5. Amide 12 derived from N-succinimidyl ester 9 of deoxycholic acid and (1R,2R)-1-phenyl-2-amino-1,3-propanediol 2, found to be active against Cryptococcus neoformans and the amide 17 obtained from N succinimidyl ester 9 of deoxycholic acid and (1S,2S)-1-para-nitrophenyl-2-amino 1,3-propanediol 5, is found to be potent against various gram-positive bacteria. PMID- 14741288 TI - Essential structural features of acetogenins: role of hydroxy groups adjacent to the bis-THF rings. AB - The presence of two hydroxy groups adjacent to the THF ring(s) is a common structural feature of natural acetogenins. To elucidate the role of each hydroxy group in the inhibitory action of acetogenins, we synthesized three acetogenin analogues which lack either or both of the hydroxy groups, and investigated their inhibitory activities with bovine heart mitochondrial complex I. Our results indicate that the presence of either of the two hydroxy groups sufficiently sustains a potent inhibitory effect. PMID- 14741289 TI - Omega-carboxypyridyl substituted ureas as Raf kinase inhibitors: SAR of the amide substituent. AB - Bis-aryl ureas have been disclosed previously as a potent class of Raf kinase inhibitors. Modifications in the amide portion led to an improvement in aqueous solubility, an important characteristic for an oral drug. Based on this finding, we hypothesize that this portion of the molecule is directed towards the solvent in Raf-1. PMID- 14741290 TI - N-4-Substituted-benzyl-N'-tert-butylbenzyl thioureas as vanilloid receptor ligands: investigation on the role of methanesulfonamido group in antagonistic activity. AB - A series of N-4-substituted-benzyl-N'-tert-butylbenzyl thioureas were prepared for the study of their agonistic/antagonistic activities to the vanilloid receptor in rat DRG neurons. Their structure-activity relationship reveals that not only the two oxygens and amide hydrogen of sulfonamido group, but also the optimal size of methyl in methanesulfonamido group play an integral role for the antagonistic activity on vanilloid receptor. PMID- 14741291 TI - Discovery of thiophene-2-carboxylic acids as potent inhibitors of HCV NS5B polymerase and HCV subgenomic RNA replication. Part 1: Sulfonamides. AB - The discovery of a novel class of HCV NS5B polymerase inhibitors, 3 arylsulfonylamino-5-phenyl-thiophene-2-carboxylic acids is described. SAR studies have yielded several potent inhibitors of HCV polymerase as well as of HCV subgenomic RNA replication in Huh-7 cells. PMID- 14741292 TI - Discovery of thiophene-2-carboxylic acids as potent inhibitors of HCV NS5B polymerase and HCV subgenomic RNA replication. Part 2: tertiary amides. AB - Further SAR studies on the thiophene-2-carboxylic acids are reported. These studies led to the identification of a series of tertiary amides that show inhibition of both HCV NS5B polymerase in vitro and HCV subgenomic RNA replication in Huh-7 cells. Structural insights about the bioactive conformation of this class of molecules were deduced from a combination of modeling and transferred NOE (trNOE) studies. PMID- 14741293 TI - Synthesis of oligonucleotide 2'-conjugates via amide bond formation in solution. AB - An efficient method for synthesis of 2'-O-carboxymethyl oligonucleotides is described. Fully deprotected oligonucleotides containing a carboxymethyl group at the 2'-position of sugar residue were obtained by a two-step procedure by periodate cleavage of an oligonucleotide containing 1,2-diol group followed by oxidation of the 2'-aldehyde resulted with sodium chlorite. 2'-O-Carboxymethyl oligonucleotides prepared were efficiently coupled in aqueous solution in the presence of a water-soluble carbodiimide to a number of amino acid derivatives or short peptides to afford novel 2'-conjugates of high purity in good yield. The method is thus shown to be suitable in principle for preparation of oligonucleotide-peptide conjugates containing an amide linkage between the 2' carboxy group of a modified oligonucleotide and the amino terminus of a peptide. PMID- 14741294 TI - Discovery of novel aspartyl ketone dipeptides as potent and selective caspase-3 inhibitors. AB - The discovery of a series of potent, selective and reversible dipeptidyl caspase 3 inhibitors are reported. The iterative discovery process of using combinatorial chemistry, parallel synthesis, moleculare modelling and structural biology will be discussed. PMID- 14741295 TI - The design and synthesis of sulfonamides as caspase-1 inhibitors. AB - A series of sulfonamides (1) has been prepared as inhibitors of interleukin-1beta converting enzyme (ICE), also known as caspase 1. These compounds were designed to improve potency by rigidifying the enzyme bound molecule through an intramolecular hydrogen bond. An X-ray crystal structure of a representative member of this series bound to the active site of ICE, confirms the presence of the hydrogen bonding interaction. PMID- 14741296 TI - 3-trifluoromethyl-4-nitro-5-arylpyrazoles are novel K(ATP) channel agonists. AB - This communication describes the discovery and synthesis of a series of 3 trifluoromethyl-4-nitro-5-arylpyrazoles as potent K(ATP) channel agonists. The most potent compound reported is ca. 100-fold more potent than diazoxide and exhibits selectivity for the SUR1 K(ATP) channel subtype. The 4-nitro substitutent on the pyrazole ring was required for activity, and limited SAR suggests that the de-protonated pyrazole maybe the active species. PMID- 14741297 TI - Synthesis and SAR evaluation of 1,2,4-triazoles as A(2A) receptor antagonists. AB - The synthesis and in vitro structure-activity relationships (SAR) of a series of triazoles as A(2A) receptor antagonists is reported. This resulted in the identification of potent, selective and permeable 1,2,4-triazoles such as 42 for further optimization and evaluation in vivo. PMID- 14741298 TI - Measurement of balance in survivors of acute lymphoblastic leukemia in childhood. AB - Survivors of acute lymphoblastic leukemia (ALL) in childhood have been identified with balance problems. The goals of this study were to objectively quantify these observations by measuring the displacement and velocity of the body centre of pressure, a measure of 'sway'. The subjects included 79 ALL survivors who were at least 1 year post-treatment for ALL and 83 age-matched controls. Testing was carried out using various static balance tests designed to challenge central nervous system maintenance of balance and postural control. Patients were different from controls, across all age groups, in the more difficult task in which they were required to balance over a narrow base of support. However this did not reach statistical significance. When they were asked to perform this task with their eyes closed 32% of the patients tested, as opposed to 2% of controls, were unable to complete the task (significant at P<0.01, chi(2)). The youngest group of patients (children aged 5-7 years) presented consistently with lower (or similar) displacement and lower velocity measures. This phenomenon has been observed in healthy children, however, in patients, these trends were magnified and beyond the normal age range in healthy children. The velocity values were significantly different from the other patient groups (normalized values for velocity; P<0.01). These observations then manifested as higher sway values when patients were challenged with a difficult task with their eyes closed. We propose that survivors of childhood ALL have delayed motor development compared to age matched controls giving rise to impairment in balance and postural control. It is possible that these impairments are a consequence of intensive multi-agent chemotherapy and cranial irradiation. PMID- 14741299 TI - Postural control: toe-standing versus heel-toe standing. AB - Toe-standing is observed in a number of populations who are able to stand without loss of balance and also those who have balance problems. Intuitively, individuals who stand on their toes are able to successfully regulate their whole body movement in order to keep themselves stable. Force platform data were collected for ten able-bodied subjects during three quiet standing postures, (a) heel-toe standing; (b) half-toe standing and (c) standing en demi pointe (full toe-standing). Differences in control mechanisms with each posture were compared using stabilogram diffusion analysis. During open-loop control (short-term), toe standing is less stable than heel-toe standing (P<0.05). There is greater stochastic activity when toe-standing (P<0.05), suggesting that any short-term instability is being compensated for by an increase in muscle activity across the lower joints. During closed-loop control (long-term), there is no difference in mediolateral (ML) stochastic activity (increased activity has been linked to falls) between toe-standing and heel-toe standing. In addition, toe-standing is more stable than heel-toe standing (P< s0.05). Toe-standing, in and of itself, may not be responsible for balance problems in populations who compulsorily toe stand. PMID- 14741300 TI - The role of the human cerebellum in short- and long-term habituation of postural responses. AB - The aim of the present study was to investigate the role of the human cerebellum in short-term (STH) and long-term habituation (LTH) of postural responses to repeated platform perturbations. Ten cerebellar patients and ten age- and sex matched healthy controls participated. Twenty backward platform translations were applied on each of 5 consecutive days. Changes of postural response size within each day were assessed to determine STH and changes across days to determine LTH. Both controls and cerebellar patients showed a significant reduction of postural response size within each day (i.e. STH). No significant reduction of postural response size was observed across days (i.e. no LTH). Both controls and cerebellar patients, however, showed a tendency of response size to increase across days suggesting long-term sensitization. The amount of changes within and across days did not significantly differ between groups. The present findings suggest that changes of postural response size to repeated perturbations do not depend upon the integrity of the cerebellum. PMID- 14741301 TI - Recovery from perturbations during paced walking. AB - The aim of the current study was to develop a safe, standardized, stability test and to explore a set of metrics to characterize the recovery of gait stability in healthy individuals following a single mechanical perturbation during steady locomotion. Balance perturbations were mechanically applied to the right foot of 12 healthy subjects during paced walking by translating a platform embedded in a 12 m walkway diagonally (+45/-135 degrees ) relative to the direction of travel approximately 200 ms after heel strike. We examined the medio-lateral (ML) displacement of the sternum before, during and after the platform translation. Measurements of ML position of the right and left shanks in relation to the position of the sternum were used as step-by-step estimates of the moment arm controlling ML motion of the body. We hypothesized that when gait is perturbed in the single stance phase of the step cycle via a translation of the support surface, a series of steps after the perturbation input will be altered reflecting an effort by the CNS to maintain the center of mass (COM) within the base of support and to stabilize the upper body for continued gait. Specifically, if the foot is perturbed laterally during mid-stance a widening of the upcoming step will occur and if the foot is perturbed medially a narrowing of the upcoming step will occur. This behavior was frequent for most subjects. Recovery of non perturbation behavior was achieved on the third step after the platform translation. An additional strategy was seen for some subjects during lateral perturbation inputs. Instead of widening the upcoming step, these subjects acquired the support to stabilize the body by putting their left foot down very quickly with minimal change in stance width. The recovery profiles of the sternum, though directionally asymmetric, were similar in shape among subjects and roughly proportional to the magnitude of the platform translation. Five to six steps were required for complete recovery in the subjects tested in this study. PMID- 14741302 TI - Sensitivity of the results produced by the inverse dynamic analysis of a human stride to perturbed input data. AB - The results of the inverse dynamic procedures used in gait analysis are known to be highly dependent on the quality of the kinematic and dynamic input data and on the biomechanical model anatomical data. In this paper the sensitivities of the system response to imprecision in the input data and biomechanical model were calculated. It was shown that the gait analysis results were very sensitive to the identification of the point of application of the external forces. The quality of the results was less sensitive to errors made during motion reconstruction and to uncertainties in the biomechanical anatomical data. In this study it is also shown that the adopted inverse dynamic analysis method, based on natural coordinates, effectively shielded any error made on a particular kinematic chain from propagation to other branches of the biomechanical model. PMID- 14741303 TI - Muscle activity during gait initiation in normal elderly people. AB - The purpose of this study was to describe the patterns of phasic muscle during gait initiation in normal elderly people. Bilateral surface EMG recordings were made of tibialis anterior, medial gastrocnemius and gluteus medius activity throughout gait initiation in 21 subjects. A variable expression of the onset muscle pattern is shown, with a tendency for muscle activity to be more variable in the preparatory phase. These results provide a baseline of normal gait initiation muscle activity against which to compare that of patients with gait initiation and balance difficulties. PMID- 14741305 TI - Joint moment control of mechanical energy flow during normal gait. AB - The study purpose was to estimate the ability of joint moments to transfer mechanical energy through the leg and trunk during gait. A segmental power analysis of five healthy adult subjects revealed that internal joint extensor moments removed energy from the leg and added energy to the trunk, while flexor moments and gravity produced the opposite effects. The only exception to this pattern was during the push off phase of gait when the ankle plantar flexor moment added energy to both the leg and the trunk. Pairs of joint moments with opposite energetic effects (knee extensor vs gravity, hip flexor vs ankle plantar flexor) worked together to balance energy flows through the segments. This intralimb coordination suggests that moments with contradictory effects are generated simultaneously to control mechanical energy flow within the body during walking. PMID- 14741304 TI - Foot force direction control during leg pushes against fixed and moving pedals in persons post-stroke. AB - The component of foot force generated by muscle action (F(m)) during pedaling in healthy humans has a nearly constant direction with increasing force magnitude. The present study investigated the effect of stroke on the control of foot force. Ten individuals with hemiparesis secondary to a cerebral vascular accident performed pushing efforts against translationally fixed and moving pedals on a custom stationary cycle ergometer. We found that while F(m) direction remained constant with increasing effort in both the fixed- and moving-crank conditions for both limbs, the orientation of that force component differed between limbs. The non-paretic limb produced the same F(m) orientation as seen previously in healthy humans. However, relative to the non-paretic limb, the paretic limb force line-of-action was shifted away from the hip and closer to the knee in the sagittal-plane for both pedal motion conditions. In the frontal plane, the paretic limb force line-of-action was shifted laterally, closer to parallel to the midline, for both pedal motion conditions. These shifts were consistent with previously reported lower limb muscle weakness and alterations in muscle activation observed during pedaling tasks following stroke. The finding of similar orientations for static and dynamic pushing efforts suggests that limb posture could be a trigger for relative muscle activation levels. The preservation of a constant direction in F(m) with increasing force magnitude post stroke, despite an orientation shift, suggests that control of lower limb force may be organized by magnitude and direction and that these two aspects are differentially affected by stroke. PMID- 14741306 TI - Validity and comparisons of tibiofemoral orientations and displacement using a femoral tracking device during early to mid stance of walking. AB - First, this study compares tibiofemoral motion during walking using a new femoral tracking device (FTD) and bone mounted markers in a single subject (n=1). The results suggest errors of <3 degrees in tibiofemoral angles using the FTD method over the first 85% of stance. Second, this study compares tibiofemoral angles and displacement during walking using the FTD method and a modified Helen Hayes method to track the femur in 13 subjects (n=13). The results suggest similar tibiofemoral angles in the sagittal and frontal planes using the two methods (average root mean square (RMS) differences <3.6+/-1.5 degrees ), and a large decrease in the transverse plane angles (average RMS differences=6.5+/-1.9 degrees ) and estimates of tibiofemoral displacement (P<0.05) using the FTD method. The FTD method presents a practical alternative to recording tibiofemoral transverse plane angles and displacement over the first 85% of stance. PMID- 14741307 TI - Use of the normalcy index for the evaluation of gait pathology. AB - The normalcy index (NI) has been proposed as a method for quantifying the amount of deviation in a subject's gait, compared to the gait of the average unimpaired person. The NI was computed for a sample of 144 children affected by cerebral palsy, five idiopathic toe-walkers and 12 able-bodied subjects. It was sensitive enough to distinguish unimpaired subjects from idiopathic toe-walkers and to distinguish between the plegic and uninvolved limbs of hemiplegic patients. The NI was robust enough to categorize pathology, ranging from mild disorders to quadriplegia. The NI was found to be clinically applicable, reliable and easy to use, making it a valuable element in the quantitative evaluation of gait pathology. PMID- 14741308 TI - Temporal patterns of plantar pressures and lower-leg muscle activity during walking: effect of speed. AB - Plantar pressure assessment is a tool useful for study of the gait cycle. In this study, we present a means of assessing the gait cycle using a temporal analysis of plantar pressures and lower-leg muscle activities. Plantar pressures and surface electromyography (EMG) of the tibialis anterior (TA) and medial gastrocnemius (MG) muscles were recorded as 19 men walked on a treadmill at seven speeds between 0.45 and 1.79 m/s. A typical 'heel strike to toe off' gait pattern was observed. Speed had minimal effects on the shapes of the muscle EMG root-mean square-and plantar pressure-time curves except for the pressure-time curves in the heel and midfoot. A linear relationship was found between speed and peak pressures in the heel, medial forefoot, and toes; pressures in these regions increased by 91-289% going from 0.45 to 1.79 m/s. The temporal pressure changes in the forefoot and toes were paralleled by changes in MG muscle activity (i.e., cross-correlations of > or =0.90); TA muscle activity was not cross-correlated with the temporal pressure patterns in any region. However, the peak values of TA muscle activity were found to be highly correlated across speeds with peak pressures in the heel and toes (i.e., r > or =0.98); similar high correlations were found between peak values of MG muscle activity and heel pressure. In summary, these data collected on able-bodied persons during motorized treadmill walking can be useful for comparison to those of patients undergoing treadmill evaluations for atypical gait cycle patterns and for tracking the progress of patients during gait rehabilitation. PMID- 14741309 TI - A functional MRI study of simple arithmetic--a comparison between children and adults. AB - The purpose of this study was to examine brain areas involved in simple arithmetic, and to compare these areas between adults and children. Eight children (four girls and four boys; age, 9-14 years) and eight adults (four women and four men; age, 40-49 years) were subjected to this study. Functional magnetic resonance imaging (fMRI) was performed during mental calculation of addition, subtraction, and multiplication of single digits. In each group, the left middle frontal, bilateral inferior temporal and bilateral lateral occipital cortices were activated during each task. The adult group showed activation of the right frontal cortex during addition and multiplication tasks, but the children group did not. Activation of the intraparietal cortex was observed in the adult group during each task. Although, activation patterns were slightly different among tasks, as well as between groups, only a small number of areas showed statistically significant differences. The results indicate that cortical networks involved in simple arithmetic are similar among arithmetic operations, and may not show significant changes in the structure during the second decade of life. PMID- 14741310 TI - Object-centred neglect for non-verbal visual stimuli. AB - In a first experiment, 11 neglect patients repeatedly bisected the elongated caricature of a basset hound with head on the right and tail on the left side with respect to the viewer. On the last (critical) trial, in which the figure was left-right reversed, the bisection error towards the ipsilesional side reversed its direction in three patients and significantly decreased in one patient. In a second experiment, 13 different neglect patients had to bisect the elongated caricature of the basset hound with head on the left and tail on the right side. On the last trial, the bisection error reversed its direction in three patients and significantly decreased in three patients. These results suggest that object centred neglect (OCN) may affect newly established knowledge about the canonical orientation of a non-verbal visual stimulus. PMID- 14741311 TI - Language-specific memory traces of consonants in the brain. AB - This study examined whether experience with a native language affected processing of a place-of-articulation feature. In Experiment 1, 10 stimuli from a continuum of synthesized Hindi bilabial, dental and retroflexed stops were presented to English and Hindi speakers to examine discrimination and identification across the bilabial/dental and dental/retroflexed boundaries. In an oddball task designed to elicit mismatch negativity (MMN), subjects ignored these stimuli while their brain activity was recorded. All participants showed similar behavioral discrimination and identification. However, the English subjects were slower that the Hindi subjects on the discrimination task. All subjects were less accurate and slower at identifying and discriminating the dental and retroflexed compared to the bilabial sounds. A small MMN was observed for some of the bilabial-dental contrasts, but not for the dental-retroflexed contrasts. No group differences were found. In Experiment 2, MMN was observed to a greater stimulus difference (bilabial-retroflexed) and was earlier when the bilabial rather than the retroflexed sound served as the frequent stimulus for both groups. The MMN was also earlier for the Hindi than the English groups when the retroflexed sound served as the frequent stimulus. These results indicate that the Hindi speakers used detailed acoustic-phonetic information for more rapid brain discrimination than the English participants and that the dental-retroflexed discrimination is more difficult than the bilabial-dental discrimination for all speakers. PMID- 14741312 TI - Dissociable neural correlates for familiarity and recollection during the encoding and retrieval of pictures. AB - Results from behavioral studies have supported the idea that recognition memory can be supported by at least two different processes, recollection and familiarity. However, it remains unclear whether these two forms of memory reflect neurally distinct processes. Furthermore, it is unclear whether recollection and familiarity can be best conceived as differing primarily in terms of retrieval processing, or whether they additionally differ at encoding. To address these issues, we used event-related brain potentials (ERPs) to monitor neural correlates of familiarity and recollection at both encoding and retrieval. Participants studied pictures of objects in two types of study blocks and subsequently made remember-know and source memory judgments during retrieval. Results showed that, during encoding, neural correlates of subsequent familiarity and recollection onsetted in parallel, but exhibited differences in scalp topography and time course. Subsequent familiarity-based recognition was associated with a left-lateralized enhanced positivity and observed at anterior scalp sites from 300 to 450 ms, whereas subsequent recollection was associated with a topographically distinct right-lateralized positivity at anterior scalp sites from 300 to 450 ms and bilateral activity from 450 to 600 ms. During retrieval, neural correlates of familiarity emerged earlier than correlates of recollection. Familiarity was associated with an enhanced positivity at frontopolar scalp sites from 150 to 450 ms, whereas recollection was associated with positive ERP modulations over bilateral frontal (300-600 ms) and parietal (450-800 ms) sites. These results demonstrate that familiarity and recollection reflect the outcome of neurally distinct memory processes at both encoding and retrieval. PMID- 14741313 TI - Comparison of blind and sighted participants' performance in a letter recognition working memory task. AB - The performance in letter recognition of 21 blind participants was compared with that of 16 age-matched sighted participants in an n-back working memory task. Blind participants were tested tactually with series of raised letters and Braille characters, and sighted participants tactually with series of raised letters and visually with series of letters presented on a computer screen. With this approach, we wanted to compare the tactual performances by trained (blind) and non-trained (sighted) participants, and tactual and visual performances by trained (blind and sighted, respectively) participants. Increments of mnemonic load increased the number of incorrect responses significantly in all n-back tasks. As expected, the blind participants outperformed the sighted ones statistically significantly in the tactile raised letters n-back task. The sighted participants produced significantly fewer incorrect responses in the visual task than the blind participants in the raised letters task, whereas there was no such difference between sighted subjects' visual performance and blind subjects' tactile performance in the Braille task. These results demonstrate the degree and limits to which everyday practice develops perceptual skills either in persons without sensory deficits (vision and visual environment), or persons with severe sensory loss (blindness and tactile environment). The performance level of blind persons relying on their tactile skills is just about the same as that of sighted subjects relying on their visual skills. PMID- 14741314 TI - Prefrontal-cingulate activation during executive control: which comes first? AB - The Stroop test requires executive control functions, in particular inhibition of a learned routine (in this case, word reading). The spatiotemporal analysis of brain activation during Stroop task execution was performed in 16 healthy subjects using high-density event-related potentials (ERPs) and dipole source modeling (BESA software). Scalp ERP analysis revealed the neurophysiological substrate of the interference effect: first, a greater negativity in the incongruent as compared to the congruent and neutral conditions was found between 350 and 450 ms poststimulus over left frontocentral scalp regions. Source analysis of the difference wave (incongruent-congruent) indicated that a generator localized in left prefrontal cortex (PFC) contributed to this effect. Second, immediately after the first effect, a greater positivity in the incongruent as compared to the congruent and neutral conditions developed between 450 and 550 ms poststimulus over midline frontocentral scalp regions. A generator of this effect was located in right anterior cingulate cortex (ACC). ACC activation seems to follow the activation of PFC with some overlap between the two components. Possible interpretation of this finding is that PFC signals ACC when executive control is required and ACC implements the control. PMID- 14741315 TI - Blue piglets? Electrophysiological evidence for the primacy of shape over color in object recognition. AB - The goal of the study was to investigate how the color and shape of visual stimuli are processed when they are conjointly presented and represent real, and familiar, entities for which normal individuals presumably have a specific 'object color knowledge' (e.g., piglets are pink, artichokes are green). There is evidence, from event related potential (ERP) literature on selective attention to color in conjunction with other, arbitrarily related, stimulus dimensions (e.g., geometrical shape), that color is processed faster than shape, and that the processing of shape depends on color relevance. In this study we recorded ERPs from 28 scalp sites in right-handed volunteers performing selective attention tasks to either color or shape of pictures representing familiar objects and animals. The results revealed that the selection of color was faster, and probably less demanding, than that of shape. However, it was also evidenced that the selection of color depended on object shape, but not vice versa. Indeed, in the attend-color condition, the N2 responses were significantly greater when stimulus shape was prototypically associated, rather than unassociated, with the color perceived. Topographical mapping of difference voltages identified the posterior occipito/temporal region of the left hemisphere as the possible locus of conjoined color and shape processing. Overall, the data support object-based attention models. PMID- 14741316 TI - Spatiotemporal differences between cognitive processes of spatially possible and impossible objects: a high-density electrical mapping study. AB - Differences in cognitive processing between spatially possible and impossible figures were investigated using event-related potentials (ERPs). Two types of figures with identical luminance and equivalent spatial frequency were used as visual stimuli: possible three-dimensional figures (drawn with perspective and existing in the three-dimensional world) and impossible figures (drawn with perspective but not existing in the three-dimensional world). High-density electroencephalographic recording (72 channels) was performed for analysis of ERPs accompanying perception of each figure type; amplitude differences between the conditions were considered neurophysiologic correlates to perceptual differences between possible and impossible objects. Low-resolution brain electromagnetic tomography (LORETA) was used to identify the current source related to the differences. Compared with impossible three-dimensional figures, perception of possible figures showed a significant negative potential increase in the right inferior occipitotemporal region between 350 and 389 ms of latency. The current source was localized to the right fusiform gyrus. The results suggest that right fusiform gyrus is involved in discrimination between spatially possible and impossible objects. PMID- 14741317 TI - An event-related fMRI study of the neurobehavioral impact of sleep deprivation on performance of a delayed-match-to-sample task. AB - Eighteen subjects (ages 18-35) underwent event-related functional magnetic resonance imaging (efMRI) while performing a delayed-match-to-sample (DMS) task before and immediately after 48 h of sustained wakefulness. The DMS trial events were: a 3-s study period of either a one-, three-, or six-letter visual array; a 7-s retention interval; and a 3-s probe period, where a button press indicated whether the probe letter was in the study array. Ordinal Trend Canonical Variates Analysis (OrT CVA) was applied to the data from the probe period for trials with six-letter study lists prior to and immediately following sleep deprivation to find an activation pattern whose expression decreased with sleep deprivation in as many subjects as possible, while being present in both conditions. The first principal component of the OrT analysis identified a covariance pattern whose expression decreased as a function of sleep deprivation in 17 of 18 subjects (p<0.001). While overall expression of the pattern showed a systematic decrease with sleep deprivation, the brain regions that make up the pattern show covarying increases and decreases in activation. Regions that decreased their activation were noted in the parietal (BA 7 and 40), temporal (BA 37, 38 and 39) and occipital (BA 18 and 19) lobes; regions that increased their activation were noted in the cerebellum, basal ganglia, thalamus and the anterior cingulate gyrus (BA 32). The reduction in pattern expression with sleep deprivation for each subject was related to the change in performance on the DMS task. Subject decreases in pattern expression were correlated with reductions in recognition accuracy (p<0.05), increased intra-individual variability in reaction time (p<0.005) and increased lapsing (p<0.005). PMID- 14741318 TI - Recovery of semantic word processing in global aphasia: a functional MRI study. AB - One important issue concerning the recovery of higher cognitive functions-such as word comprehension in aphasia-is to what extent impairments can be compensated for by intact parts of the network of areas normally involved in a closely related function ("redundancy recovery"). In a previous functional MRI investigation, we were able to show that left hemispheric redundancy recovery within a distributed system of related lexical-semantic functions was the most probable basis of recovery of comprehension from transcortical sensory aphasia. The question remained, however, whether redundancy recovery may play a more general role in the recovery of comprehension after large left hemispheric lesions and severe aphasia. We had the possibility, using the same fMRI paradigm, to study seven cases with left middle cerebral artery (MCA) infarction and partial recovery of comprehension > or =6 months after presentation with global aphasia on acute assessment. Lateralization of activation did not differ significantly between patients and controls. The most consistent regions of activation included the left extrasylvian posterior temporal and the right posterior parietal cortex. Recovery of language comprehension was associated predominantly with activations in regions, which were also activated in several normal subjects. We suggest that a redundancy recovery mechanism within multiple representations of closely related functions was more important than take-over of function by previously unrelated areas (vicariation) as the basis of recovery of word comprehension in our patients in spite of extensive left hemispheric damage. We conclude that redundancy within the lexical-semantic system seems to make an important contribution to recovery of comprehension even in severe aphasia. PMID- 14741319 TI - Mismatch negativity in schizophrenia: a family study. AB - BACKGROUND: Mismatch negativity (MMN) is a measure of cortical activity that occurs in response to a change in auditory stimuli. We investigated whether MMN is a potential marker of genetic vulnerability to schizophrenia by comparing MMN in a group of patients with schizophrenia, their unaffected relatives, and controls. METHOD: There are 25 schizophrenic patients, 37 of their unaffected first-degree relatives, and 20 unrelated controls that performed the MMN task. Linear regression with robust standard errors, and accounting for correlations within families, was employed to test for differences in MMN amplitude between the groups. RESULTS: Patients had significantly smaller MMN amplitudes compared to both their unaffected relatives and controls at FZ (P<0.01) and at F3 (P=0.01), whereas relatives and controls did not differ at FZ or at F3. No differences were found between any of the groups at F4. Furthermore, we found no strong evidence that the MMN amplitude is a familial trait. CONCLUSIONS: Our results confirm that the MMN amplitude is reduced in schizophrenia. However, the MMN does not show a significant familial influence and is normal among the unaffected relatives. We conclude that while the MMN is abnormal in patients with schizophrenia, it is a weak or unreliable marker of vulnerability when applied to subclinical populations, and therefore is unlikely to be an endophenotype for the disorder. PMID- 14741320 TI - Visual scanpath dysfunction in first-degree relatives of schizophrenia probands: evidence for a vulnerability marker? AB - Previous research demonstrates that people with schizophrenia have abnormally 'restricted' visual scanpaths to face and facial expression stimuli, which appear to be diagnostically specific to schizophrenia [Schizophr. Res. 55 (2002) 159; Biol. Psychiatry 52 (2002) 338]. This study examined the familial transmission of 'restricted' scanpaths in first-degree relatives of schizophrenia subjects. We recorded visual scanpaths for 65 schizophrenia subjects, 37 biological first degree relatives and 61 nonrelated 'healthy' control subjects in two experiments: 'face recognition' and 'facial affect recognition'. Concurrent behavioral tasks were face matching and expression matching, each under two multiple-choice conditions (seven or three options). As predicted, first-degree relatives generally showed an attenuated form of the markedly 'restricted' scanpaths of schizophrenia subjects across all face stimuli. The notable exception to this pattern was the relatives' extreme avoidance of facial features (compared to both schizophrenia and healthy control groups). Our results offer the first evidence that some components of visual scanpath dysfunction may represent a trait marker in the familial transmission of schizophrenia, but that first-degree relatives may have additional disturbances in social cognition associated with the perception of facial features. PMID- 14741321 TI - Neurological and morphological anomalies and the genetic liability to schizophrenia: a composite phenotype. AB - BACKGROUND: Neurological soft signs (NSS) and minor physical anomalies (MPA) are frequent in patients with schizophrenia and their biological relatives. We examined whether the NSS and MPA are related to the genetic loading in schizophrenia. METHODS: Patients with schizophrenia (DSM-IV) (n=61), nonpsychotic parents of these patients (n=76) and healthy comparison subjects (n=44) took part in the study. Parents were further classified as "presumed carriers" of the genetic loading (n=26) if they had a second relative with schizophrenia in their ascendants and/or collaterals (first or second degree) or as "presumed noncarriers" (n=50). NSS and MPA were compared in these groups. RESULTS: A multivariate analysis indicated that total NSS and MPA scores, adjusted for age and gender, were significantly related to group status. Univariate tests showed higher scores in motor coordination and integration subscores (p=0.005 and 0.008, respectively) in presumed carriers than in presumed noncarriers. In addition, a discriminant function analysis based on total NSS and MPA scores correctly classified 71% of nonpsychotic parents in presumed carriers or presumed noncarriers. CONCLUSIONS: Neurological impairments and slight morphological anomalies seem to be associated with the genetic risk for schizophrenia, even when the disease itself is absent. Their presence might be a valuable composite intermediate phenotype for genetic studies. PMID- 14741322 TI - Normal cerebral asymmetry in familial and non-familial schizophrenic probands and their unaffected relatives. AB - Loss of normal fronto-occipital cerebral asymmetry has been reported in patients with schizophrenia and also in their well relatives from multiply affected families, suggesting a relationship with susceptibility genes. We sought to confirm this relationship in a family study of patients with schizophrenia and their unaffected relatives of presumed differing genetic risk. MRI scans were carried out on 25 probands from families multiply affected with the disorder, and 36 of their unaffected relatives, 34 probands from families with no other affected members, 42 of their unaffected relatives, and 76 controls. Volumetric measurements of prefrontal, premotor, sensorimotor and occipitoparietal regions were obtained from which a measure of fronto-occipital torque was derived. There were no significant differences in measurements of fronto-occipital torque between the subject groups. Both schizophrenic probands and their relatives displayed the normal pattern of cerebral asymmetry, with larger right than left frontal regions and a larger left than right occipitoparietal region. Our findings failed to confirm an association between loss of fronto-occipital torque and genetic liability for schizophrenia and also failed to replicate the previously reported association between loss/reversal of fronto-occipital asymmetry and schizophrenia. PMID- 14741323 TI - Microarray screening of lymphocyte gene expression differences in a multiplex schizophrenia pedigree. AB - In order to help prioritize the selection of candidate genes and to study possible trait and not state related changes in gene expression, we compared lymphocytic gene expression patterns of five individual family members with schizophrenia and nine unaffected individuals from a large multiplex high density pedigree. We screened gene expression by microarray consisting of 1128 brain focused genes. Three criteria for selection of microarray gene differences between schizophrenia and unaffected family members were employed: a significant t-test, expression in a majority of subjects, and fold change magnitude. Gene expression levels were significantly different for nine genes between individuals with schizophrenia compared to unaffected controls, and two genes were validated by real-time PCR. The expression of the neuropeptide Y receptor Y1 gene (NPY1R localized at 4q31.3-q32) and the human guanine nucleotide-binding regulatory protein Go-alpha (GNAO1 localized at 16q13) was significantly decreased in individuals with schizophrenia compared to unaffected family controls by microarray and real-time PCR. The cytosolic malate dehydrogenase gene (MDH1 localized at 2p13.3) was also significantly increased by microarray analysis and showed a trend for increase by real-time PCR. The significant genes are discussed in terms of proximity to linkage regions, prior association studies of schizophrenia, and other reports of microarray screening of schizophrenia tissue. Evidence from these studies taken together with the present study suggests critical pathways in schizophrenia may be studied in peripheral tissue as part of the strategy in functional genomic convergence. This preliminary study needs to be repeated by screening a larger set of genes in additional families with schizophrenia. The present study offers support for examination of gene expression patterns using lymphocytic RNA for complex neuropsychiatric disorders from large cohorts of patients. PMID- 14741324 TI - Meta-analysis of association between the T102C polymorphism of the 5HT2a receptor gene and schizophrenia. AB - A meta-analysis of whole-genome linkage scans confirmed linkage between schizophrenia and markers on the long arm of chromosome 13. The gene HTR2A, which codes for the 5HT2a receptor, is located in this area. The T102C single nucleotide polymorphism of HTR2A has been the subject of much research. The production of the C-allele form of HTR2A is significantly less than that of the T allele form in normal controls and schizophrenic patients. Although the association of schizophrenia with the C allele of HTR2A was confirmed by a meta analysis 5 years ago, there was a continuous debate because negative findings were also considerable, which may have been due to ethnic differences in association. We performed another meta-analysis, since the number of available studies of this association has recently doubled. In the meta-analysis of 31 case control association studies, we found a significant association between the C allele of the T102C polymorphism and schizophrenia, which was more pronounced in European samples than in the entire sample. We found significant heterogeneity in the allele-wise analysis (C vs. T) and homozygous genotype-wise analysis (CC vs. TT), both of which were at least partially explained by differences between samples from Asian and European countries. In East Asian countries, there was not a significant association with the C allele or CC homozygosity, indicating strong genetic differences and noncombinability of data between European and East Asian populations. Interestingly, the frequency of the T allele was much higher in East Asian patients and controls (59.5% and 57.5%, respectively) than in European patients and controls (40% and 43.5%, respectively). In five family-based association studies, we did not find significant evidence for association of the C allele with schizophrenia; yet, the pooled OR was 1.3 (95% CI=0.9-1.8, z=1.47, p=0.14), which is consistent with the results of the case-control studies. The effects of other genes, environmental effects on DNA methylation, or different methods of classification may be the causes for such heterogeneity, but more study in this area is needed. PMID- 14741325 TI - Risperidone response and 5-HT6 receptor gene variance: genetic association analysis with adjustment for nongenetic confounders. AB - Previous genetic-response studies, usually without considering environmental factors, encountered great difficulties in replication of results. Although atypical antipsychotics are becoming the mainstay for schizophrenia treatment which makes an antipsychotic "atypical" remains unclear. Risperidone (a widely used atypical antipsychotic agent) and several other atypicals have high affinities for 5-HT6 and 5-HT7 receptors. This study investigated the effects of the T-->C 267 polymorphism in the 5HT6 receptor gene and two rare Pro279Leu and Thr92Lys substitutions in the 5HT7 receptor gene on risperidone efficacy after rigorous control for nongenetic confounders. We found an association between the T-->C 267 polymorphism of the 5HT6 receptor gene and response to risperidone in 123 acutely ill schizophrenia inpatients after adjustment for confounders. Compared to patients with the T/C 267 genotype, those with T/T 267 showed less severe positive symptoms (p=0.006) and general psychopathology (including anxiety, depression, and cognitive dysfunctions) (p=0.005). The T-->C 267 polymorphism had no influences on negative symptoms. The two rare polymorphisms in the 5HT7 receptor gene were not observed in our sample. In conclusion, the 5HT6 receptor gene variant can affect risperidone response to positive symptoms and general psychopathology (but not negative symptoms) after control for nongenetic factors. PMID- 14741326 TI - Differences in p53 gene polymorphisms between Korean schizophrenia and lung cancer patients. AB - The reduced incidence of cancer observed in schizophrenia patients may be related to differences in genetic background. It has been suggested that genetic predisposition towards schizophrenia is associated with reduced vulnerability to lung cancer, and p53 gene is one of the candidate genes. We tested the genetic association between schizophrenia and lung cancer by analyzing polymorphic sites in the p53 gene. Genotype and allele frequencies at two polymorphic sites in the p53 gene (BstUI and MspI restriction sites in exon 4 and intron 6, respectively) were studied in Korean schizophrenia (n=179) and lung cancer patients (n=104). Comparisons of the genotype and allele frequencies of the MspI polymorphism revealed significant differences between schizophrenia and lung cancer patients. The results suggest that the p53 polymorphism specifically found in schizophrenia patients may be associated with reduced vulnerability to lung cancer. PMID- 14741327 TI - The 3' region of the DRD2 gene is involved in genetic susceptibility to schizophrenia. AB - The gene coding for the D2 dopamine receptor (DRD2) is considered as one of the most relevant candidate genes in schizophrenia. Previous genetic studies focusing on this gene yielded conflicting results, for example because of differences in methodology (linkage versus association studies) and variability in the loci analyzed (the DRD2 gene having many polymorphic sites). We used a progressive strategy with two different approaches (case-control and transmission disequilibrium test) and investigated six genetic polymorphisms spanning the DRD2 gene in 103 patients with DSM-IV criteria of schizophrenia, their 206 parents and 83 matched healthy control subjects. We found a significant excess of the A2 allele in subject with schizophrenia compared to unaffected controls. An excess of transmission of the A2 allele (and haplotypes containing this marker) from the parents to the affected children was also observed. Interestingly, the TaqI A1/A2 polymorphism, located 9.5 kb downstream from the DRD2 gene, maps in a novel gene, untitled "X-kinase", and leads to a 713Glu-->Lys substitution in exon 8. As the analysis of the other markers within the DRD2 gene does not improve the strength of the association, our data are in favor of a specific role of the 3' chromosomic region of the DRD2 gene in the vulnerability to schizophrenia. PMID- 14741328 TI - Do the siblings of schizophrenia patients demonstrate affect perception deficits? AB - Affect perception deficits have been extensively documented in schizophrenia and are associated with the social dysfunction that is characteristic of this disorder. The two previous studies examined facial affect perception in genetically at-risk samples and yielded mixed results. The current study was designed to provide a rigorous test of affect perception abilities among schizophrenia patients (n=58), their biological siblings without psychosis (n=51), and nonpsychiatric controls (n=49). Participants completed three measures of affect perception, including facial, vocal, and combined modality. Schizophrenia patients performed significantly worse than controls on two of the three affect perception tests as well as a composite index based on all three tests. The performance of the sibling group fell between the patient and control groups on each of the affect perception tests. However, group differences achieved statistical significance only for the composite index with the siblings performing significantly worse than controls and significantly better than the schizophrenia group. These findings demonstrate that subtle deficits in affect perception are detectable in the unaffected siblings of schizophrenia patients when multiple measures of different types of affect perception abilities are used in combination. PMID- 14741329 TI - Epigenetic studies of genomic retroelements in major psychosis. AB - This work is dedicated to the exploration of the role of epigenetic (epiG) factors in major psychosis. One of the key functions of epigenetic modification of the genome of eukaryotic cells is to suppress transcriptional activity of the retroelements. Examples of retroelements are endogenous retroviral sequences (ERVs), Alu's, and LINEs, among others, which as a rule are hypermethylated. There is evidence from schizophrenia (SCH) and other human complex diseases that some of the genomic retroelements become transcribed in the affected tissues. Our goal was to screen DNA samples from post-mortem brain tissues of individuals who were affected with major psychiatric illness for retroelements that were located in the hypomethylated fraction of the genomic DNA. Over 100 Alu sequences were cloned, sequenced, and mapped to the human genome. A substantial portion of the cloned Alu's are located close to or within the genes that may be interesting targets for further genetic, transcription, and epigenetic studies. PMID- 14741330 TI - Family-based association study of the functional monoamine oxidase A gene promoter polymorphism and schizophrenia. PMID- 14741331 TI - Failure to find association between PRODH deletion and schizophrenia. PMID- 14741332 TI - Microcystin-LR and okadaic acid-induced cellular effects: a dualistic response. AB - Microcystins, potent heptapeptide hepatotoxins produced by certain bloom-forming cyanobacteria, are strong protein phosphatase inhibitors. They covalently bind the serine/threonine protein phosphatases 1 and 2A (PP1 and PP2A), thereby influencing regulation of cellular protein phosphorylation. The paralytic shellfish poison, okadaic acid, is also a potent inhibitor of these PPs. Inhibition of PP1 and PP2A has a dualistic effect on cells exposed to okadaic acid or microcystin-LR, with both apoptosis and increased cellular proliferation being reported. This review summarises the existing data on the molecular effects of microcystin-LR inhibition of PP1 and PP2A both in vivo and in vitro, and where possible, compares this to the action of okadaic acid. PMID- 14741333 TI - Snake and snail toxins acting on nicotinic acetylcholine receptors: fundamental aspects and medical applications. AB - This review covers recent data on interactions of nicotinic acetylcholine receptors (AChR) with snake venom proteins (alpha- and kappa-neurotoxins, 'weak' toxins recently shown to act on AChRs), as well as with peptide alpha-conotoxins from Conus snails. Mutations of AChRs and toxins, X-ray/nuclear magnetic resonance structures of alpha-neurotoxin bound to AChR fragments, and the X-ray structure of the acetylcholine-binding protein were used by several groups to build models for the alpha-neurotoxin-AChR complexes. Application of snake toxins and alpha-conotoxins for pharmacological distinction of muscle, neuronal and neuronal-like AChR subtypes and for other medical purposes is briefly discussed. PMID- 14741334 TI - Mitochondrial effectors in caspase-independent cell death. AB - Activation of caspases is recognized as a key element in the apoptotic process. However, new evidence is drawing attention to the emergent role of cell death pathways where caspases are not involved. Recent advances in the molecular understanding of these new ways to die, called caspase-independent, have revealed that mitochondria play an important role via the release of proapoptotic proteins. The purpose of this review is to integrate, from a biological and structural point of view, the most recent advances in the knowledge of the main mitochondrial proapoptotic proteins involved in this cell death cascade. The origin of programmed cell death is discussed through these strongly conserved effectors. PMID- 14741335 TI - Effective accumulation of polyion complex micelle to experimental choroidal neovascularization in rats. AB - Exudative age-related macular degeneration, characterized by choroidal neovascularization (CNV), is a major cause of visual loss. In this study, we examined the distribution of the polyion complex (PIC) micelle encapsulating FITC P(Lys) in blood and in experimental CNV in rats to investigate whether PIC micelle can be used for treatment of CNV. We demonstrate that PIC micelle has long-circulating characteristics, accumulating to the CNV lesions and is retained in the lesion for as long as 168 h after intravenous administration. These results raise the possibility that PIC micelles can be used for achieving effective drug targeting to CNV. PMID- 14741336 TI - Role of the PLA2-independent peroxiredoxin VI activity in the survival of immortalized fibroblasts exposed to cytotoxic oxidative stress. AB - Peroxiredoxin VI (PrxVI) is a bifunctional enzyme with non-selenium glutathione peroxidase and Ca2+-independent acidic phospholipase A2 activities. We demonstrate that transfection-mediated PrxVI overexpression protects immortalized human WI-38 and murine NIH3T3 fibroblasts against cytotoxic doses of tert butylhydroperoxide and H2O2. Mutants for either glutathione peroxidase or phospholipase A2 activity show that glutathione peroxidase but not phospholipase A2 activity is required to promote cell survival after stress. Also, ectopic PrxVI overexpression does not protect telomerase-stabilized WI-38 fibroblasts against stress-induced premature senescence. PMID- 14741337 TI - Chalcone dimethylallyltransferase from Morus nigra cell cultures. Substrate specificity studies. AB - A new prenyltransferase (PT) enzyme derived from the microsomal fractions of cell cultures of Morus nigra was shown to be able to prenylate exclusively chalcones with a 2',4'-dihydroxy substitution and the isoflavone genistein. Computational studies were performed to shed some light on the relationship between the structure of the substrate and the enzymatic activity. PT requires divalent cations, particularly Mg(2+), to be effective. The apparent K(m) values for gamma,gamma-dimethylallyldiphosphate and 2',4'-dihydroxychalcone were 63 and 142 microM, respectively. The maximum activity of the enzyme was expressed during the first 10 days of cell growth. PMID- 14741338 TI - Expression and secretion of human apolipoprotein A-I in the heart. AB - Various studies have correlated apolipoprotein (apo) A-I, the major component high-density lipoprotein, with protection against development of cardiovascular disease. Although apoA-I expression has been previously detected in the liver and intestine, we have discovered that the human apoA-I gene is also expressed in the heart. Using transgenic (Tg) mice generated with the human apoA-I/C-III/A-IV gene cluster and Tg mice produced with just the 2.2 kb human apoA-I gene, we have detected significant levels of apoA-I expression in the heart. Furthermore, the detection of apoA-I expression in the hearts of human apoA-I Tg mice indicates that the minimal regulatory elements necessary for cardiac expression of the gene are located near its coding sequence. To determine if the apoA-I gene is also expressed in the human heart, similar analyses were performed, where apoA-I expression was found in both adult and fetal hearts. Furthermore in-depth investigation of the various regions of human and Tg mouse hearts revealed that the apoA-I mRNA was present in the ventricles and atria, but not in the aorta. In situ hybridization of Tg mouse hearts revealed that apoA-I expression was restricted to the cardiac myocyte cells. Finally, heart explants and cardiac primary culture experiments with Tg mice showed secretion of particles containing the human apoA-I protein, and metabolic labeling experiments have also detected a 28 kDa human apoA-I protein secreted from the heart. From these novel findings, new insights into the role and function of apoA-I can be extrapolated. PMID- 14741339 TI - Bacillus subtilis YxaG is a novel Fe-containing quercetin 2,3-dioxygenase. AB - The Bacillus subtilis genome contains genes for three hypothetical proteins belonging to the bicupin family, two of which we have previously shown to be Mn(II)-dependent oxalate decarboxylases. We have now shown that the third, YxaG, exhibits quercetin 2,3-dioxygenase activity and that it contains Fe ions. This contrasts with the eukaryotic enzyme which contains a Cu ion. YxaG is the first prokaryotic carbon monoxide-forming enzyme that utilises a flavonol to be characterised and is only the second example of a prokaryotic dioxygenolytic carbon monoxide-forming enzyme known to contain a cofactor. It is proposed to rename the B. subtilis gene qdoI. PMID- 14741340 TI - Structural characteristics and refolding of in vivo aggregated hyperthermophilic archaeon proteins. AB - Several recombinant proteins in inclusion bodies expressed in Escherichia coli have been measured by Fourier transform infrared and solid-state nuclear magnetic resonance spectra to provide the secondary structural characteristics of the proteins from hyperthermophilic archaeon Pyrococcus horikoshii OT3 (hyperthermophilic proteins) in inclusion bodies. The beta-strand-rich single chain Fv fragment (scFv) and alpha-helix-rich interleukin (IL)-4 lost part of the native-like secondary structure in inclusion bodies, while the inclusion bodies composed of the hyperthermophilic proteins of which the native form is alpha helix rich, are predominated by alpha-helix structure. Further, the secondary structure of the recombinant proteins solubilized from inclusion bodies by detergent or denaturant was observed by circular dichroism (CD) spectra. The solubilization induced the denaturation of the secondary structure for scFv and IL-4, whereas the solubilized hyperthermophilic proteins have retained the alpha helix structure with the CD properties resembling those of their native forms. This indicates that the hyperthermophilic proteins form native-like secondary structure in inclusion bodies. Refolding of several hyperthermophilic proteins from in vivo aggregated form without complete denaturation could be accomplished by solubilization with lower concentration (e.g. 2 M) of guanidine hydrochloride and removal of the denaturant via stepwise dialysis. This supports the existence of proteins with native-like structure in inclusion bodies and suggests that non native association between the secondary structure elements leads to in vivo aggregation. We propose a refolding procedure on the basis of the structural properties of the aggregated archaeon proteins. PMID- 14741341 TI - The interaction between E-tropomodulin and thymosin beta-10 rescues tumor cells from thymosin beta-10 mediated apoptosis by restoring actin architecture. AB - Thymosin beta-10 (TB10) is a small G-actin binding protein that induces depolymerization of intracellular F-actin pools by sequestering actin monomers. Previously, we demonstrated that overexpression of TB10 in ovarian tumor cells increased the rate of cell death. As an initial step to define molecular mechanism of TB10-dependent apoptotic process in ovarian tumor cells, we searched a human ovary cDNA library for a novel TB10 binding protein using a yeast two hybrid system. The selected protein was human E-tropomodulin (E-Tmod), another component of the actin binding proteins. Subsequently, two interacting protein components were determined quantitatively. Results showed that the full-length TB10 is required to bind with E-Tmod, and the TB10 binding site on E-Tmod partially overlaps with the actin binding site on E-Tmod. Moreover, introduction of E-Tmod cDNA into a tumor cell line reversed TB10 mediated apoptosis and restored actin architectures. These results may suggest that TB10 regulates apoptotic homeostasis by not only just binding to actin but also competing or blocking the protein complex formation of E-Tmod with actin. PMID- 14741342 TI - Increased leptin and white adipose tissue hypoplasia are sexually dimorphic in Lif null/Igf-I haploinsufficient mice. AB - We previously showed cooperation of leukemia inhibitory factor (LIF) and insulin like growth factor I (IGF-I) during development. Mice doubly deficient in LIF and IGF-I died at birth. We now analyze the possible combined influence of both factors on postnatal growth. The haploinsufficiency of the Igf-I gene on a Lif null background caused a marked reduction in body mass index and white adipose tissue only in female mice. These animals had increased leptin, increased serum IGF-I and apparent substitution of white adipose tissue by brown adipose tissue. The complex interrelationships between LIF and IGF-I in regulating weight thus involve sexually dimorphic effects on adipose tissue differentiation and circulating leptin. PMID- 14741343 TI - Oxidative refolding of lysozyme in trifluoroethanol (TFE) and ethylene glycol: interfering role of preexisting alpha-helical structure and intermolecular hydrophobic interactions. AB - The oxidative refolding of equilibrium intermediates of lysozyme stabilized in trifluoroethanol (TFE) and ethylene glycol was monitored. Equilibrium intermediates of disulfide reduced lysozyme in TFE are known to contain considerable amounts of alpha-helical structure and resemble the early intermediate in the oxidative refolding of lysozyme. We find that the intermediates in TFE do not proceed to folding; they form aggregates. However, interestingly, intermediates in ethylene glycol refold to the native state with improved folding yield. Secondary structure of these intermediates was monitored by far-UV circular dichroism. Our results indicate that formation of alpha helical structure prior to oxidative refolding does not help the process in the case of lysozyme. Interfering with intermolecular hydrophobic interactions in the unfolded state is more productive. PMID- 14741344 TI - Hepatitis B virus X protein modulates peroxisome proliferator-activated receptor gamma through protein-protein interaction. AB - Ligand activation of peroxisome proliferator-activated receptor gamma (PPARgamma) has been reported to induce growth inhibition and apoptosis in various cancers including hepatocellular carcinoma (HCC). However, the effect of hepatitis B virus X protein (HBx) on PPARgamma activation has not been characterized in hepatitis B virus (HBV)-associated HCC. Herein, we demonstrated that HBx counteracted growth inhibition caused by PPARgamma ligand in HBx-associated HCC cells. We found that HBx bound to DNA binding domain of PPARgamma and HBx/PPARgamma interaction blocked nuclear localization and binding to recognition site of PPARgamma. HBx significantly suppressed a PPARgamma-mediated transactivation. These results suggest that HBx modulates PPARgamma function through protein-protein interaction. PMID- 14741345 TI - Structural studies on delta(3)-delta(2)-enoyl-CoA isomerase: the variable mode of assembly of the trimeric disks of the crotonase superfamily. AB - Subunits of the enzymes in the crotonase superfamily form tight trimeric disks. In most members of this protein superfamily these disks assemble further into hexamers. Here we report on the 2.1 A structure of a tight hexameric crystal form of the yeast peroxisomal delta(3)-delta(2)-enoyl-CoA isomerase (Eci1p). A comparison of this structure to a previously solved crystal form of Eci1p and other structures of this superfamily shows that there is much variability with respect to the relative distance between the disks and their relative orientations. In particular helices H2 and H9 are involved in the inter-trimer contacts and there are considerable structural differences in these helices in this superfamily. Helices H2 and H9 are near the catalytic cavity and it is postulated that the observed structural variability of these helices, stabilized by the different modes of assembly, has allowed the evolution of the wide range of substrate and catalytic specificity within this enzyme superfamily. PMID- 14741346 TI - CRP2 is an autonomous actin-binding protein. AB - Cysteine-rich proteins (CRPs) have been shown to be involved in cell differentiation, transcriptional regulation and the organisation of the actin cytoskeleton. Thus far, the latter function has been inferred solely from the in vitro interaction of CRP1, CRP2, and CRP3 with alpha-actinin and zyxin. We show here that purified, recombinant CRP2 binds directly to F-actin in vitro in co sedimentation assays. Using a green fluorescent protein (GFP)-tagged construct of CRP2 we analysed its localisation and dynamics in A7r5 rat smooth muscle cells. CRP2 was associated with the actin cytoskeleton and decorated actin stress fibres in a continuous fashion, unlike the periodic labelling pattern observed for alpha actinin and zyxin, which also accumulate in focal adhesions. Using live video fluorescence microscopy we observed the behaviour of GFP-CRP2 during the dynamic rearrangement of the actin cytoskeleton in phorbol 12,13-dibutyrate-treated A7r5 cells. In contrast to the actin-binding proteins SM22alpha and alpha-actinin, GFP CRP2 did not translocate into the podosomes induced by this treatment, but remained preferentially bound to the stress fibres, suggesting an actin filament stabilising role for CRP2. When fused to the mitochondrial targeting sequence from the Listeria protein ActA, GFP-CRP2 was almost completely localised to mitochondria, but no significant recruitment of either alpha-actinin or zyxin could be observed. Taken together, our results demonstrate that CRP2 can bind to F-actin directly and that the association with the actin cytoskeleton is independent of alpha-actinin or zyxin localisation in the cell, thus questioning the role of CRP2 as a regulator of alpha-actinin function in vivo. PMID- 14741347 TI - Placental growth factor induces FosB and c-Fos gene expression via Flt-1 receptors. AB - Placental growth factor (PlGF) is a member of the vascular endothelial growth factor (VEGF) family that binds specifically to Flt-1. The biological roles of PlGF and Flt-1 have not yet been defined and the signalling mechanisms mediating cellular actions of PlGF remain poorly understood. In human umbilical vein endothelial cells, VEGF and PlGF induced expression of both full-length FosB mRNA and an alternatively spliced variant, DeltaFosB, with similar efficacy and kinetics. In contrast, PlGF induced c-Fos expression less strongly than VEGF, and whereas VEGF strongly upregulated tissue factor mRNA, PlGF had a negligible effect. PlGF induced c-Fos expression in porcine aortic endothelial cells specifically expressing Flt-1, and FosB expression in the monocytic RAW 264.7 cell line expressing endogenous Flt-1. These findings show for the first time that VEGF and PlGF induce mRNA expression of the transcription factors FosB and c Fos, and suggest that these factors may play a role in the biological responses mediated by PlGF and Flt-1. PMID- 14741348 TI - Cation-selective ion channels formed by p7 of hepatitis C virus are blocked by hexamethylene amiloride. AB - A 63 residue peptide, p7, encoded by hepatitis C virus was synthesised and tested for ion channel activity in lipid bilayer membranes. Ion channels formed by p7 had a variable conductance: some channels had conductances as low as 14 pS. The reversal potential of currents flowing through the channels formed by p7 showed that they were permeable to potassium and sodium ions and less permeable to calcium ions. Addition of Ca(2+) to solutions made channels formed by p7 less potassium- or sodium-selective. Hexamethylene amiloride, a drug previously shown to block ion channels formed by Vpu encoded by HIV-1, blocked channels formed by p7. In view of the increasing number of peptides encoded by viruses that have been shown to form ion channels, it is suggested that ion channels may play an important role in the life cycle of many viruses and that drugs that block these channels may prove to be useful antiviral agents. PMID- 14741349 TI - Functional and structural characterization of a novel member of the natriuretic family of peptides from the venom of Pseudocerastes persicus. AB - A novel peptide, PNP (Pseudocerastes persicus natriuretic peptide), was isolated from the venom of the Iranian viper P. persicus. Amino acid sequencing revealed that the 37-residue peptide belongs to the family of natriuretic peptides. The physiological effects of intra-venously PNP infused into anesthetized rats on urine flow, sodium excretion and blood pressure were comparable to those of atrial natriuretic peptide (ANP). In PC12 cells that were treated with either PNP, ANP, or C-type natriuretic peptide, PNP induced a similar cGMP response as ANP. Since PC12 cells only express the natriuretic peptide receptor (NPR)-A receptor we conclude that PNP binds to the NPR-A receptor. The solution conformation of PNP was characterized using (1)H nuclear magnetic resonance spectroscopy and indicates a high degree of conformational flexibility. PMID- 14741350 TI - A plant outer mitochondrial membrane protein with high amino acid sequence identity to a chloroplast protein import receptor. AB - We have identified a novel protein on the outer membrane of Arabidopsis thaliana mitochondria. This protein displays 67% sequence identity with the 64 kDa translocase of the outer envelope membrane of chloroplasts (Toc). A mitochondrial localisation for this protein was determined by (i). its presence in the proteome of highly purified Arabidopsis mitochondria, (ii). Western blot analysis with antibodies to Toc64 from pea that indicate its presence in Arabidopsis and pea mitochondria, (iii). green fluorescent protein fusion proteins that indicate an exclusive mitochondrial localisation for this protein, and (iv). expression profiles in various tissue types and during development that are more similar to translocase of the outer mitochondrial membrane components than to chloroplastic Toc components. Thus Arabidopsis mitochondria contain a protein with high sequence identity to a plastid protein import receptor. PMID- 14741351 TI - Investigations by mass isotopomer analysis of the formation of D-2 hydroxyglutarate by cultured lymphoblasts from two patients with D-2 hydroxyglutaric aciduria. AB - D-2-Hydroxyglutaric aciduria is an inborn error of metabolism first described in 1980. To date, more than 40 patients have been diagnosed with this disease. To identify the metabolic precursor of D-2-hydroxyglutarate (D-2-HG), cultured human lymphoblasts from two patients with D-2-HG aciduria were grown in culture medium supplemented with [U-(13)C(6)]glucose or [(2)H(5)]glutamate. Mass isotopomer distribution measurements of D-2-HG, 2-ketoglutarate (2-KG) and citrate were performed by gas chromatography-mass spectrometry. The mass isotopomer distributions in D-2-HG, 2-KG and citrate, following [U-(13)C(6)]glucose and [(2)H(5)]glutamate incubations, revealed that 2-KG interconverts rapidly to D-2 HG and that D-2-HG is formed within the mitochondria. PMID- 14741352 TI - Quantitative measurements of Ca(2+)/calmodulin binding and activation of myosin light chain kinase in cells. AB - Myosin II regulatory light chain (RLC) phosphorylation by Ca(2+)/calmodulin (CaM) dependent myosin light chain kinase (MLCK) is implicated in many cellular actin cytoskeletal functions. We examined MLCK activation quantitatively with a fluorescent biosensor MLCK where Ca(2+)-dependent increases in kinase activity were coincident with decreases in fluorescence resonance energy transfer (FRET) in vitro. In cells stably transfected with CaM sensor MLCK, increasing [Ca(2+)](i) increased MLCK activation and RLC phosphorylation coincidently. There was no evidence for CaM binding but not activating MLCK at low [Ca(2+)](i). At saturating [Ca(2+)](i) MLCK was not fully activated probably due to limited availability of cellular Ca(2+)/CaM. PMID- 14741353 TI - Absence of mechanical allodynia and Abeta-fiber sprouting after sciatic nerve injury in mice lacking membrane-type 5 matrix metalloproteinase. AB - Matrix metalloproteinases (MMPs) are a family of endopeptidases that degrade extracellular matrix components. Membrane-type 5 MMP (MT5-MMP/MMP-24) was identified as neuron-specific, and is believed to contribute to neuronal circuit formation and plasticity. To elucidate its function in vivo, we have generated mice lacking MT5-MMP by gene targeting. MT5-MMP-deficient mice were born without obvious morphological abnormalities. No apparent histological defects were observed in the nervous system either. However, MT5-MMP-deficient mice did not develop neuropathic pain with mechanical allodynia after sciatic nerve injury, though responses to acute noxious stimuli were normal. Neuropathic pain induced by peripheral nerve lesions is known to accompany structural reorganization of the nervous system. Intraneural injection of cholera toxin B subunit, a transganglionic tracer, into the injured sciatic nerve of wild-type mice revealed that the myelinated Abeta-fiber primary afferents sprouted from laminae III-VI of the dorsal horn of the spinal cord and invaded lamina II. However, no such sprouting and invasion of Abeta-fibers were observed in MT5-MMP-deficient mice. These findings suggest that MT5-MMP is essential for the development of mechanical allodynia and plays an important role in neuronal plasticity in this mouse model. PMID- 14741354 TI - TRF2 is in neuroglial cytoplasm and induces neurite-like processes. AB - TRF2 is a ubiquitous protein that protects telomeres in the nucleus. We found that TRF2 was present at the peripheral nerve axons and the brain neuroglial cell processes extensively. It was in the cytoplasmic membrane as well as nuclear fractions, but not in the soluble cytoplasmic fraction of SH-SY5Y neuroblastoma cells. TRF2 was up-regulated in P19 embryonal carcinoma cells at the early stage of induced neural differentiation with retinoic acid treatment. Upon transfection, TRF2-expressing COS cells often produced neurite-like long cytoplasmic processes. TRF2 is a component of neuroglial cells and appears to be involved in the cytoplasmic process formation that is necessary for neural differentiation. PMID- 14741355 TI - In vivo formation of glutamyl-tRNA(Gln) in Escherichia coli by heterologous glutamyl-tRNA synthetases. AB - Two types of glutamyl-tRNA synthetase exist: the discriminating enzyme (D-GluRS) forms only Glu-tRNA(Glu), while the non-discriminating one (ND-GluRS) also synthesizes Glu-tRNA(Gln), a required intermediate in protein synthesis in many organisms (but not in Escherichia coli). Testing the capacity to complement a thermosensitive E. coli gltX mutant and to suppress an E. coli trpA49 missense mutant we examined the properties of heterologous gltX genes. We demonstrate that while Acidithiobacillus ferrooxidans GluRS1 and Bacillus subtilis Q373R GluRS form Glu-tRNA(Glu), A. ferrooxidans and Helicobacter pylori GluRS2 form Glu tRNA(Gln) in E. coli in vivo. PMID- 14741356 TI - Chronological aging-independent replicative life span regulation by Msn2/Msn4 and Sod2 in Saccharomyces cerevisiae. AB - Mutations in RAS2, CYR1, and SCH9 extend the chronological life span in Saccharomyces cerevisiae by activating stress-resistance transcription factors and mitochondrial superoxide dismutase (Sod2). Here we show that mutations in CYR1 and SCH9 also extend the replicative life span of individual yeast mother cells. However, the triple deletion of stress-resistance genes MSN2/MSN4 and RIM15, which causes a major decrease in chronological life span, extends replicative life span. Similarly, the overexpression of superoxide dismutases, which extends chronological survival, shortens the replicative life span and prevents budding in 30-40% of virgin mother cells. These results suggest that stress-resistance transcription factors Msn2/Msn4 negatively regulate budding and the replicative life span in part by increasing SOD2 expression. The role of superoxide dismutases and of other stress-resistance proteins in extending the chronological life span of yeast, worms, and flies indicates that the negative effect of Sod2, Msn2/Msn4/Rim15 on the replicative life span of S. cerevisiae is independent of aging. PMID- 14741357 TI - Immunoseparation of Prion protein-enriched domains from other detergent-resistant membrane fractions, isolated from neuronal cells. AB - The possibility of coexistence of different subtypes of membrane lipid rafts has been investigated in cerebellar granule cells, by submitting detergent-resistant membrane fractions to immunoprecipitation. Among the proteins and lipids present in detergent-resistant fractions, almost all Prion protein, GAP43 and PKC were present in the immunoprecipitate obtained with anti-GAP43 or anti-Prion protein antibody at 4 degrees C, together with a small fraction of cholesterol and sphingolipids, suggesting that they belong to a distinct subset of membranes. On the contrary, all Fyn and almost all MARCKS remained in the supernatant. Fluorescence microscopy experiments showed that Fyn and Prion protein were mostly not colocalized within a single neuron. Our results suggest that granule cells membranes contains different subtypes of detergent-resistant fractions, possibly deriving from different lipid rafts. PMID- 14741358 TI - MG-63 human osteosarcoma cells grown in monolayer and as three-dimensional tumor spheroids present a different metabolic profile: a (1)H NMR study. AB - High resolution proton nuclear magnetic resonance ((1)H NMR) spectroscopy was used to determine if the same cell line (MG-63 human osteosarcoma cells) grown in monolayer or as small (about 50-80 microm in diameter), three-dimensional tumor spheroids with no hypoxic center has different metabolic characteristics. Consequently, the (1)H NMR spectra were obtained from both types of cultures and then compared. The results indicate that the type of cellular spatial array determines specific changes in MG-63 cells. In particular, small but significant differences in lactate and alanine indicating a perturbation in energy metabolism were observed in the two cell models. In addition, although variations in CH(2) and CH(3) groups were also seen, it is not possible at this time to establish if lipid metabolism is truly different in cells and spheroids. PMID- 14741359 TI - Apoptin's functional N- and C-termini independently bind DNA. AB - Apoptin induces apoptosis specifically in tumour cells, where Apoptin is enriched in the DNA-dense heterochromatin and nucleoli. In vitro, Apoptin interacts with dsDNA, forming large nucleoprotein superstructures likely to be relevant for apoptosis induction. Its N- and C-terminal domains also have cell-killing activity, although they are less potent than the full-length protein. Here, we report that both Apoptin's N- and C-terminal halves separately bound DNA, indicating multiple independent binding sites. The reduced cell killing activity of both truncation mutants was mirrored in vitro by a reduced affinity compared to full-length Apoptin. However, none of the truncation mutants cooperatively bound DNA or formed superstructures, which suggests that cooperative DNA binding by Apoptin is required for the formation of nucleoprotein superstructures. As Apoptin's N- and C-terminal fragments not only share apoptotic activity, but also affinity for DNA, we propose that both properties are functionally linked. PMID- 14741360 TI - The endoproteolytic processing of alphavbeta5 integrin is involved in cytoskeleton remodelling and cell migration. AB - We previously showed that the post-translational cleavage of alphav subunit is essential for integrin-dependent signalling and cell adhesion. Here, we report that blocking alphav subunit cleavage by expression of alpha1-PDX, a convertase inhibitor, modified the capacity of cells to change shape, via a remodelling of the actin cytoskeleton upon cell attachment. These changes are associated with cell scattering and with a dramatic increase in cell migration to vitronectin. The alphav subunit cleavage is thus essential for integrin function and has a considerable impact on integrin-dependent events, especially those leading to cell migration. PMID- 14741361 TI - A new member of the bacterial ribonuclease inhibitor family from Saccharopolyspora erythraea. AB - We have identified Sti, the gene of a ribonuclease inhibitor from Saccharopolyspora erythraea, by using a T7 phage display system. A specific phage has been isolated from a genome library by a biopanning procedure, using RNase Sa3, a ribonuclease from Streptomyces aureofaciens, as bait. Sti, a protein of 121 amino acid residues, with molecular mass 13059 Da, is a homolog of barstar and other microbial ribonuclease inhibitors. To overexpress its gene in Escherichia coli, we optimized the secondary structure of its mRNA by introducing a series of silent mutations. Soluble protein was isolated and purified to homogeneity. Inhibition constants of complex of Sti and RNase Sa3 or barnase were determined at pH 7 as 5 x 10(-12) or 7 x 10(-7), respectively. PMID- 14741362 TI - Effective reduction of antigenicity of hen egg lysozyme by site-specific glycosylation. AB - Various mutant lysozymes were constructed by genetic modification and secreted in yeast expression system to evaluate the changes in the antigenicity of hen egg lysozyme (HEL). Although Arg68, the most critical residue to antigenicity of HEL, was substituted with Gln, the binding of monoclonal antibodies (mAbs) with the mutant lysozyme did not critically reduce, remaining 60% of the binding with mAb. In contrast, glycosylated mutant lysozyme G49N whose glycine was substituted with asparagine dramatically reduced the binding with mAb. The oligomannosyl type of G49N lysozyme reduced binding with mAb to one-fifth, while the polymannosyl type of G49N lysozyme completely diminished the binding with mAb. This suggests that the site-specific glycosylation of lysozyme in the interfacial region of lysozyme antibody complex is more effective to reduce the antigenicity than the mutation of single amino acid substitution in the interfacial region. PMID- 14741363 TI - Biophysical characterization of hepatitis C virus core protein: implications for interactions within the virus and host. AB - A primary function of the hepatitis C virus (HCV) core protein is to package the viral genome within a nucleocapsid. In addition, core protein has been shown to interact with more than a dozen cellular proteins, and these interactions have been suggested to play critical roles in HCV pathogenesis. A more complete knowledge of the biophysical properties of the core protein may help to clarify its role in HCV pathogenesis and nucleocapsid assembly and provide a basis for the development of novel anti-HCV therapies. Here we report that recombinant mature core protein exists as a large multimer in solution under physiological conditions. Far-UV circular dichroism (CD) experiments showed that the mature core protein contains stable secondary structure. Studies with truncated core protein demonstrated that the C-terminal region of the core protein is critical for its folding and oligomerization. Intrinsic fluorescence spectroscopy and near UV CD analysis indicated that the tryptophan-rich region (residues 76-113) is largely solvent-exposed and not likely responsible for multimerization of the mature core protein in vitro. PMID- 14741364 TI - The contribution of serine residues 1588 and 1755 to phosphorylation of the type I inositol 1,4,5-trisphosphate receptor by PKA and PKG. AB - Type I inositol 1,4,5-trisphosphate receptors can be phosphorylated by cAMP dependent protein kinase (PKA) and cGMP-dependent protein kinase (PKG). To define the site-specificity of these events we analyzed the phosphorylation of mutant receptors expressed in intact cells. These studies showed that S(1588) and S(1755), the serine residues within kinase consensus sequences, are equally sensitive to PKA, that phosphorylation events at these sites are independent of each other, and that PKG predominantly phosphorylates S(1588). These findings provide the basis for understanding the functional consequences of type I inositol 1,4,5-trisphosphate receptor phosphorylation. PMID- 14741365 TI - Impas 1 possesses endoproteolytic activity against multipass membrane protein substrate cleaving the presenilin 1 holoprotein. AB - Presenilins (PS1 and PS2) are supposed to be unusual aspartic proteases and components of the gamma-secretase complex regulating cleavage of type I proteins. Multiple mutations in PS1 are a major cause of familial early-onset Alzheimer's disease (AD). We and others recently identified PS-related families of proteins (IMPAS/PSH/signal peptide peptidases (SPP)). The functions of these proteins are yet to be determined. We found that intramembrane protease-associated or intramembrane protease aspartic protein Impas 1 (IMP1)/SPP induces intramembranous cleavage of PS1 holoprotein in cultured cells coexpressing these proteins. Mutations in evolutionary invariant sites in hIMP1 or specific gamma secretase inhibitors abolish the hIMP1-mediated endoproteolysis of PS1. In contrast, neither AD-like mutations in hIMP1 nor in PS1 substrate abridge the PS1 cleavage. The data suggest that IMP1 is a bi-aspartic polytopic protease capable of cleaving transmembrane precursor proteins. These data, to our knowledge, are a first observation that a multipass transmembrane protein or the integral protease per se may be a primary substrate for an intramembranous proteolysis. PMID- 14741366 TI - Enhancement of RNAi activity by improved siRNA duplexes. AB - RNA interference (RNAi) is a powerful tool for suppressing the expression of a gene of interest, in which 21-25 nucleotide short interfering RNA (siRNA) duplexes homologous to the silenced gene function as sequence-specific RNAi mediators. The present study shows that newly designed siRNA duplexes, 'fork siRNA duplexes', whose sense-stranded siRNA elements carry one to four nucleotide mismatches at the 3'-ends against the antisense-stranded siRNA elements, can enhance RNAi activity over conventional siRNA duplexes in cultured mammalian cells. PMID- 14741368 TI - Tyrosine phosphorylation of BIT on photic stimulation in the rat retina. AB - BIT is a transmembrane glycoprotein with three immunoglobulin-like domains in its extracellular region and tyrosine phosphorylation sites in its cytosolic region. We have previously shown that BIT was tyrosine phosphorylated in the hypothalamic suprachiasmatic nucleus in response to light exposure during the dark period, and suggested that it was involved in the light entrainment of the circadian clock. To further investigate the function of BIT in the nervous system, we examined the effect of photic stimulation on its tyrosine phosphorylation in the rat retina. It was found that the tyrosine phosphorylation level of BIT in the retina was higher in the light period than in the dark period. In addition, a light stimulation during the dark period resulted in a rapid phosphorylation of BIT and a subsequent association of BIT with SHP-2. The phosphorylation state was quickly reverted when the light was turned off. The light-dependent phosphorylation of BIT was also observed in isolated cultured retinas, and this was blocked by a specific Src-family inhibitor, PP-2. Immunohistochemical study showed that BIT was highly enriched in the inner and outer plexiform layers in the retina, where the immunoreactivity to anti-SHP-2 antibody was also detected. These results suggest that tyrosine phosphorylation of BIT is involved in neuronal transmission in the retina. PMID- 14741367 TI - Two Arabidopsis thaliana genes encode functional pectin methylesterase inhibitors. AB - We have identified, expressed and characterized two genes from Arabidopsis thaliana (AtPMEI-1 and AtPMEI-2) encoding functional inhibitors of pectin methylesterases. AtPMEI-1 and AtPMEI-2 are cell wall proteins sharing many features with the only pectin methylesterase inhibitor (PMEI) characterized so far from kiwi fruit. Both Arabidopsis proteins interact with and inhibit plant derived pectin methylesterases (PMEs) but not microbial enzymes. The occurrence of functional PMEIs in Arabidopsis indicates that a mechanism of controlling pectin esterification by inhibition of endogenous PMEs is present in different plant species. PMID- 14741369 TI - NIRF is a ubiquitin ligase that is capable of ubiquitinating PCNP, a PEST containing nuclear protein. AB - We previously reported the association of a novel Np95/ICBP90-like RING finger protein (NIRF) with a novel PEST-containing nuclear protein (PCNP). NIRF is a nuclear protein with a ubiquitin-like domain, a PHD finger, a YDG/SRA domain, Rb binding motifs and a RING finger. In this study, we showed that NIRF has auto ubiquitination activity, the hallmark of a ubiquitin ligase. PCNP was readily ubiquitinated in 293 and COS-7 cells, and NIRF ubiquitinated PCNP in vitro as well as in vivo. Considering that NIRF is implicated in cell cycle regulation, these findings suggest that NIRF and PCNP are a ubiquitin ligase and its substrate, respectively, and may constitute a novel signaling pathway with some relation to cell proliferation. PMID- 14741370 TI - Mitochondrial functional interactions between frataxin and Isu1p, the iron-sulfur cluster scaffold protein, in Saccharomyces cerevisiae. AB - Friedreich's ataxia is caused by a deficit in the mitochondrial protein frataxin. The present work demonstrates that in vivo yeast frataxin Yfh1p and Isu1p, the mitochondrial scaffold protein for the Fe-S cluster assembly, have tightly linked biological functions, acting in concert to promote the Fe-S cluster assembly. A synthetic lethal screen on high iron media with the mild G107D yfh1 mutant has specifically identified Isu1p. Analysis of the cellular phenotypes resulting from pairwise combinations of yfh1 and isu1 mutations, and cross-linking experiments in isolated mitochondria provide evidence for a direct interaction between Yfh1p and Isu1p. PMID- 14741371 TI - Blockade of v-Src-stimulated tumor formation by the Src homology 3 domain of Crk associated substrate (Cas). AB - Crk-associated substrate (Cas) is highly phosphorylated by v-Src and plays a critical role in v-Src-induced cell transformation. In this study, we found that the Src homology (SH) 3 domain of Cas blocked v-Src-stimulated anchorage independent cell growth, Matrigel invasion, and tumor growth in nude mice. Biochemical analysis revealed that the Cas SH3 domain selectively inhibited v-Src stimulated activations of AKT and JNK, but not ERK and STAT3. Attenuation of the AKT pathway by the Cas SH3 domain rendered v-Src-transformed cells susceptible to apoptosis. Inhibition of the JNK pathway by the Cas SH3 domain led to suppression of v-Src-stimulated invasion. Taken together, our results indicate that the Cas SH3 domain has an anti-tumor function, which severely impairs the transforming potential of v-Src. PMID- 14741372 TI - Evolutional analysis of human influenza A virus N2 neuraminidase genes based on the transition of the low-pH stability of sialidase activity. AB - The 1957 and 1968 human pandemic influenza A virus strains as well as duck viruses possess sialidase activity under low-pH conditions, but human H3N2 strains isolated after 1968 do not possess such activity. We investigated the transition of avian (duck)-like low-pH stability of sialidase activities with the evolution of N2 neuraminidase (NA) genes in human influenza A virus strains. We found that the NA genes of H3N2 viruses isolated from 1971 to 1982 had evolved from the side branches of NA genes of H2N2 epidemic strains isolated in 1968 that were characterized by the low-pH-unstable sialidase activities, though the NA genes of the 1968 pandemic strains preserved the low-pH-stable sialidase. These findings suggest that the prototype of the H3N2 epidemic influenza strains isolated after 1968 probably acquired the NA gene from the H2N2 low-pH-unstable sialidase strain by second genetic reassortment in humans. PMID- 14741373 TI - Chondroitin sulfate of appican, the proteoglycan form of amyloid precursor protein, produced by C6 glioma cells interacts with heparin-binding neuroregulatory factors. AB - Appican produced by rat C6 glioma cells, the chondroitin sulfate (CS) proteoglycan form of the amyloid precursor protein, contains an E disaccharide, GlcUA-GalNAc(4,6-O-disulfate)-, in its CS chain. In this study, the appican CS chain from rat C6 glioma cells was shown to specifically bind several growth/differentiation factors including midkine (MK) and pleiotrophin (PTN). In contrast, the appican CS from SH-SY5Y neuroblastoma cells contained no E disaccharide and showed no binding to either MK or PTN. These findings indicate that the E motif is essential in the interaction of the appican CS chain with growth/differentiation factors, and suggest that glial appican may mediate the regulation of neuronal cell adhesion and migration and/or neurite outgrowth. PMID- 14741374 TI - Evidence that cleavage of the precursor enzyme by autocatalysis caused secretion of multiple amylases by Aspergillus niger. AB - The observation that a mutant strain of Aspergillus niger isolated for protease overproduction accumulated Taka-amylase supported an earlier report that processing of the precursor amylase by protease resulted in the secretion of multiple amylases. Studies using a mutant strain revealed that such processing was not due to aspergillopepsin but to autocatalysis by an inherent protease activity of the precursor and glucoamylase. Alignment of protease sequences with glucoamylase showed regions of consensus with serine carboxypeptidase of A. niger. Thus point mutations in this region due to ultraviolet radiation apparently caused the mutant to evolve with enhanced protease activity that degraded the precursor and accumulated Taka-amylase. PMID- 14741375 TI - Creatine increases IGF-I and myogenic regulatory factor mRNA in C(2)C(12) cells. AB - Addition of creatine to the differentiation medium of C(2)C(12) cells leads to hypertrophy of the myotubes. To investigate the implication of insulin-like growth factor I (IGF-I) and myogenic regulatory factors (MRFs) in this hypertrophy, their mRNA levels were assessed during the first 72 h of differentiation. Creatine significantly increased the IGF-I mRNA level over the whole investigated period of time, whereas the MRF mRNA levels were only augmented at precise moments, suggesting a general activation mechanism for IGF-I and a specifically regulated mechanism for MRF transcription. Our results suggest therefore that creatine-induced hypertrophy of C(2)C(12) cells is at least partially mediated by overexpression of IGF-I and MRFs. PMID- 14741376 TI - Independent folding and conformational changes of the barnase module in the VL barnase immunofusion: calorimetric evidence. AB - Although stability is critical for in vivo application of immunotoxins, a thermodynamic description of their folding/stability is still lacking. We applied differential scanning calorimetry (DSC) to RNase-based immunofusion comprising barnase, cytotoxic RNase from Bacillus amyloliquefaciens, fused to the light chain variable domain (VL) of anti-human ferritin antibody F11. By analyzing DSC curves recorded with or without preheating and addition of the barnase stabilizing ligand guanosine 3'-monophosphate, we (i). assigned two well-resolved thermal transitions to the VL and barnase modules of VL-barnase, (ii). demonstrated independent folding of these two modules, and (iii). showed altered stability of the barnase module, which resulted from the dimeric state of VL barnase. PMID- 14741377 TI - Evidence for hetero-association of transmembrane helices of integrins. AB - Integrins are important transmembrane cell-surface receptors, which mediate interactions of the cell with other cells or the extracellular matrix. Integrins are heterodimers composed of an alpha- and a beta-subunit. They can switch between different activation states depending on intra- or extracellular signals. Inside/out and outside/in signaling is mediated via integrins across the membrane. A biologically important and yet still unanswered question is the role of the transmembrane domains in the signaling event. Here it is shown by simulated annealing/molecular dynamics calculations that recently published structural data of the cytoplasmic domains of integrin alphaIIbbeta3 are supporting a structure with interacting transmembrane helices. This corroborates a model of transmembrane domains that are actively involved in the transmembrane signaling event. PMID- 14741378 TI - Antisense suppression of the Arabidopsis PIF3 gene does not affect circadian rhythms but causes early flowering and increases FT expression. AB - Photoperiodic control of flowering is regulated by light and a circadian clock. Feedback regulation of the transcription of clock components is one of the most common and important mechanisms that control clock functions in animals, fungi, and plants. The Arabidopsis circadian clock is believed to involve two myb related proteins, LHY (late elongated hypocotyl) and CCA1 (circadian clock associated 1), which negatively regulate TOC1 (timing of cab expression 1) gene expression through direct binding to the TOC1 promoter. PIF3 (phytochrome interacting factor 3), a bHLH transcription factor binds promoter regions of the LHY and CCA1 genes, affecting the light induction of these genes, and interacts with TOC1 protein. Although the positive feedback regulation of clock components in plants has been predicted, and PIF3 has been assumed to be involved, the molecular nature of this process has not been elucidated. Here we demonstrate that the antisense suppression of the PIF3 gene causes higher levels of mRNA of floral activator genes CO (constans) and FT (flowering locus T) and results in early flowering under long days (LD). Neither the circadian rhythms of the clock controlled genes (CCGs) under constant conditions nor the diurnal rhythms of the CCGs under LD conditions are affected by the reduction in PIF3 gene expression. These results suggest that PIF3 may play an important role in the control of flowering through clock-independent regulation of CO and FT gene expression in Arabidopsis. PMID- 14741379 TI - Mitochondrial small heat-shock protein enhances thermotolerance in tobacco plants. AB - To clarify the role of mitochondrial small heat-shock protein (MT-sHSP) in the heat-shock response, we introduced the tomato (Lycopersicon esculentum) MT-sHSP gene under the control of the 35S promoter into tobacco (Nicotiana tabacum), and examined the thermotolerance of the transformed plants. Irrespective of the orientation, sense or antisense, of the gene, the transgenic plants exhibited a normal morphology and growth rate in the vegetative growth stage. When 4-week-old seedlings were exposed to sudden heat stress, the sense plants which overexpress the MT-sHSP gene exhibited thermotolerance, whereas the antisense plants in which the expression of the gene is suppressed exhibited susceptibility. PMID- 14741380 TI - RORalpha1 and RORalpha4 suppress TNF-alpha-induced VCAM-1 and ICAM-1 expression in human endothelial cells. AB - Retinoic acid receptor-related orphan receptor-alpha (RORalpha) is a nuclear orphan receptor. Adenovirus-mediated overexpression of RORalpha1 and RORalpha4 suppressed tumor necrosis factor-alpha (TNF-alpha)-induced expression of vascular cell adhesion molecule-1 (VCAM-1) and intracellular adhesion molecule-1 (ICAM-1) in human umbilical vein endothelial cells. Overexpression of RORalpha1 and RORalpha4 also suppressed TNF-alpha-stimulated translocation of p50 and p65 to the nucleus. In contrast, dominant-negative deletion mutants of RORalpha1 and RORalpha4 failed to suppress the induction of VCAM-1 and ICAM-1 and translocations of p50 and p65. These results suggest that RORalpha1 and RORalpha4 regulate the inflammatory responses via inhibition of the nuclear factor-kappaB signaling pathway in endothelial cells. PMID- 14741381 TI - PKA phosphorylation and 14-3-3 interaction regulate the function of neurofibromatosis type I tumor suppressor, neurofibromin. AB - Neurofibromin, a neurofibromatosis type I (NF1) tumor suppressor gene product, has a domain acting as a GTPase activating protein and functions in part as a negative regulator of Ras. Loss of neurofibromin expression in NF1 patients is associated with elevated Ras activity and increased cell proliferation. Therefore, regulation of the function of neurofibromin is heavily involved in cell growth and differentiation. In the present study, we identified a novel cellular neurofibromin-associating protein, 14-3-3, which belongs to a highly conserved family of proteins that regulate intracellular signal transduction events in all eukaryotic cells. The interaction of 14-3-3 is mainly directed to the C-terminal domain (CTD) of neurofibromin, and the cAMP-dependent protein kinase (PKA)-dependent phosphorylation clustered on CTD-Ser (2576, 2578, 2580, 2813) and Thr (2556) is required for the interaction. Interestingly, the increased phosphorylation and association of 14-3-3 negatively regulate the function of neurofibromin. These findings indicate that PKA phosphorylation followed by 14-3-3 protein interaction may modulate the biochemical and biological functions of neurofibromin. PMID- 14741382 TI - Induction of p53-independent apoptosis by the BH3-only protein ITM2Bs. AB - The p53 tumor suppressor protein is critically involved in cell cycle regulation and programmed cell death. Here we show that expression of the BH3-only protein ITM2Bs is able to induce apoptotic cell death in p53+/+, as well as in p53-/- cell lines. This cell death involves neither subcellular redistribution of p53 nor transcriptional regulation of p53 target genes such as Bax, Ras, Puma or Bcl 2. Together, our data provide evidence for a p53-independent apoptotic role of ITM2Bs. PMID- 14741383 TI - Hydrolysis of sphingosylphosphocholine by neutral sphingomyelinases. AB - Sphingosylphosphocholine (SPC), the N-deacylated form of sphingomyelin (SM), is a naturally occurring lipid mediator. However, little is known about the metabolism of SPC. We here report an in vitro assay system for SPC-phospholipase C (PLC). Using this assay system, we demonstrated that nSMase1 and nSMase2, human neutral sphingomyelinases (SMases), are capable of hydrolyzing SPC efficiently under detergent-free conditions. Bacterial and plasmodial neutral SMases also showed SPC-PLC activity. The substrate specificity of neutral SMases that hydrolyze SM, SPC, and monoradyl glycerophosphocholine, but not diradyl glycerophosphocholine, suggested that a hydrogen-bond donor at the C-2 or sn-2 position in the substrate is required for recognition by the enzymes. PMID- 14741386 TI - Effects of running exercise during recovery from hindlimb unloading on soleus muscle fibers and their spinal motoneurons in rats. AB - The effects of hindlimb unloading and recovery with or without running exercise on morphological and metabolic properties of soleus muscle fibers and their spinal motoneurons in rats were investigated. Ten-week-old rats were hindlimb suspended for 2 weeks and thereafter were rehabilitated with or without voluntary running exercise for 2 weeks. A decreased percentage of type I fibers and atrophy of all types of fibers were observed after hindlimb unloading. In addition, decreased oxidative enzyme activity of all types of fibers was observed after hindlimb unloading. In contrast, an improvement in the decreased percentage of type I fibers, decreased fiber cross-sectional area, and decreased fiber oxidative enzyme activity was observed after recovery with running exercise, but not without running exercise. There were no changes in the number, cell body size, or oxidative enzyme activity of motoneurons innervating the soleus muscle after hindlimb unloading or recovery with or without running exercise. These results indicate that running exercise is beneficial for the recovery of the decreased percentage of type I fibers and the atrophy and decreased oxidative enzyme activity of all types of fibers in the soleus muscle induced by hindlimb unloading and that there are no changes in morphological or metabolic properties of spinal motoneurons innervating the soleus muscle following decreased or increased neuromuscular activity. PMID- 14741387 TI - Callosal effects of transcranial magnetic stimulation (TMS): the influence of gender and stimulus parameters. AB - Transcranial magnetic stimulation (TMS) of the motor cortex of one hemisphere (conditioning stimulus, CS) inhibits EMG responses evoked in distal hand muscles by a magnetic stimulus given at appropriate interval later over the opposite hemisphere (test stimulus, TS). The common interpretation attributes this effect to an inhibition produced at cortical level via a transcallosal route. The variability of cortical excitability as measured by the interhemispheric paired pulse (PP) technique has been assessed in healthy subjects in order to compare sub- and supra-threshold intensity of CS (80% versus 120% of individual motor threshold, MT). Within- and between-subject variability relating, respectively, to interhemispheric and gender differences were also assessed. Results point to an efficacy of a magnetic CS on one hemisphere in inhibiting EMG responses of the abductor digiti minimi (ADM) stimulated by a TS delivered over the opposite hemisphere in a range of intervals centered at 12ms. These reductions were produced by the 120% suprathreshold CS, while the 80% subthreshold CS did not affect EMG responses. Females showed a higher transcallosal inhibition than males, suggesting gender differences in interhemispheric connectivity that concern the anterior half of the trunk of the corpus callosum. PMID- 14741388 TI - Plasticity of spinal nicotinic acetylcholine receptors following spinal nerve ligation. AB - The nicotinic cholinergic system is known to be important in the processing of nociceptive information. In the spinal cord, nicotinic receptors are expressed on primary afferent terminals, inhibitory interneurons and descending noradrenergic and serotoninergic fibers. Following peripheral nerve injury, the expression of numerous receptors involved in nociceptive processing is altered in the superficial dorsal horn of the spinal cord. However, the expression of nicotinic acetylcholine receptor subunits in the lumbar spinal cord following peripheral nerve injury has not been investigated. We examined the expression of the alpha3, alpha4, alpha5, alpha7, beta2, beta3 and beta4 nicotinic subunits in the spinal cord of normal and spinal nerve ligated rats using immunocytochemistry. Two nicotinic subunits were found to have an increased expression following spinal nerve ligation. The number of cells expressing the alpha3 subunit in the dorsal horn increased bilaterally following spinal nerve injury. Also, the number of alpha5 immunoreactive fibers increased significantly ipsilateral to ligation. The expression of the alpha4, alpha7, beta2, beta3 and beta4 subunits was unchanged. We propose that the increased expression of the alpha3 and alpha5 nicotinic subunits may contribute to the mechanical hypersensitivity observed following spinal nerve ligation. PMID- 14741389 TI - Stimulus frequency dependence of the linear relationship between local cerebral blood flow and field potential evoked by activation of rat somatosensory cortex. AB - We investigated the relationship between evoked local cerebral blood flow (LCBF) and the field potential induced by somatosensory activation. The specific aim of the present study was to examine the correlation between variations of evoked LCBF and field potential when the stimulus duration was changed, and the dependency of the correlation on stimulus frequency. Evoked LCBF was measured using laser-Doppler flowmetry and the field potential was observed using a tungsten electrode inserted into the cortex alpha-chloralose-anesthetized rats. The cortex was activated by electrical stimulation of the hind paw with a 1.5 mA pulse (0.1 ms) applied at frequencies of 0.5, 1, 5 and 10 Hz for durations of 2, 5, 8, 10 or 15s. We extended our previous finding [Neurosci. Res. 40 (2001) 281 290], that both the magnitude of evoked LCBF (integrated LCBF) and the summed field potential (SigmaFP) exhibited a maximum at a stimulus frequency of 5 Hz to five different stimulus durations. Moreover, although variations of integrated LCBF and SigmaFP induced by changes in the stimulus duration were linearly correlated, the slope of the regression line depended on the stimulus frequency. This stimulus frequency dependence of the integrated LCBF-SigmaFP linear relationship may be because the vessel response is frequency dependent. PMID- 14741390 TI - Architecture of binocular disparity processing in monkey inferior temporal cortex. AB - Neurons in the inferior temporal (IT) cortex respond not only to the shape, color or texture of objects, but to the horizontal positional disparity of visual features in the right and left retinal images. IT neurons with similar shape selectivity cluster in columns. In this study, we examined how IT neurons are spatially arranged in the IT according to their selectivity for binocular disparity. With a single electrode, we simultaneously recorded extracellular action potentials from a single neuron and those from background multiple neurons at the same sites or recorded multineuronal responses at successive sites along electrode penetrations, while monkeys performed a fixation task. For neurons at each recording site, effective shapes were first determined from a set of 20 shapes presented at the zero-disparity plane. The most effective shape was then presented with varying amounts of disparity. Single neuron responses and background multiunit responses recorded at the same sites showed a similar ability of disparity discrimination and tended to share the preferred disparity, suggesting that neurons with similar disparity selectivity are clustered in the IT. We estimated from sequential recordings along electrode penetrations that the size of the neuronal clusters with similar disparity selectivity was smaller than the size of clusters with similar shape selectivity. PMID- 14741391 TI - In vivo calcium imaging of cerebral cortex in hypoxia-ischemia followed by developmental stage-specific injury in rats. AB - The parietal area is a part of the cortex that is vulnerable in the rat to hypoxia-ischemia (HI) within the early postnatal period. To investigate the localizing mechanism of this cortical injury, we spatiotemporally detected the cortical intracellular calcium changes, as revealed by a calcium-sensitive fluorescence dye, Rhod 2-AM, during 1h of HI on postnatal days 7-21 in vivo. The calcium level rose to different levels at different cortical points in all animals within the first 20 min. Over the whole cortical area in the camera field, the changes in three groups significantly differed across time at 30 and 60 min, and a chronic increase appeared at days 7-8. After 3h of reperfusion, microtubule-associated protein 2 (MAP-2) immunoreactivity confirmed that parietal injury was more serious at day 7, whereas the imaging of calcium distribution did not segregate the injured and uninjured areas. Our in vivo findings in the whole brain structure indicate that the age-specific vulnerability of the parietal cortex injury is affected indirectly by the chronic increase in the late HI phase in the early postnatal period, suggesting that each cortical area differs postnatally with respect to the development of calcium regulation and signal transduction involving neural cell death and/or survival. PMID- 14741392 TI - Muscle stretch-induced modulation of noxiously activated dorsal horn neurons of feline spinal cord. AB - The present work was designed to check for the possibility of interactions between mechanical innocuous and chemically induced noxious muscle afferent inputs on discharge behavior of nociceptive superficial dorsal horn neurons (SDHNs) of the spinal cord in decerebrated cats. The innocuous and noxious stimuli were applied separately and in combination, so that the effects of the innocuous stimulus on nociceptive processing could be evaluated. The innocuous stimulus consisted of ramp-and-hold stretches of the gastrocnemius muscles, whereas the noxious stimulus consisted of i.a. injections of bradykinin (BK; 0.5 1 ml, 50 microg/ml) into the arterial circulation of same muscles. Only neurons up to approximately 1mm depth and those that responded to noxious pinch of the gastrocnemius muscles were selected for further analysis. The activity of 16 dorsal horn neurons was recorded extracellularly with high-impedance glass microelectrodes, out of which seven responded to stretch, while 12 neurons responded to bradykinin injections. The bradykinin injections induced three types of responses: excitatory, inhibitory and mixed. The majority of the neurons that showed excitatory and mixed responses to bradykinin were also influenced by stretches applied directly after the bradykinin injection. In these neurons, the stretch usually counteracted the bradykinin-induced response, i.e. shortening and reducing bradykinin-induced excitation and re-exciting the cells after bradykinin induced inhibition. The mechanism of the stretch modulation is proposed to reside in a segmental spinal control of the nociceptive transmission. PMID- 14741393 TI - Involvement of highly polysialylated neural cell adhesion molecule (PSA-NCAM) positive granule cells in the amygdaloid-kindling-induced sprouting of a hippocampal mossy fiber trajectory. AB - The mossy fiber system in the hippocampus of amygdaloid-kindled rats was examined by using highly polysialylated neural cell adhesion molecule (PSA-NCAM) as a marker for immunohistochemical detection of immature dentate granule cells and mossy fibers in combination with bromodeoxyuridine (BrdU) labeling of newly generated granule cells. Statistically significant increases in BrdU-labeled cells and PSA-NCAM-positive cells occurred in the dentate gyrus following kindling. The increase in PSA-NCAM-immunoreactive neurites was confined to the entire stratum lucidum of CA3. Immunoelectron-microscopic examination also revealed that PSA-NCAM-positive immature synaptic terminals of the sprouting mossy fibers increased in the stratum lucidum of CA3 in the kindled rats. The increase in the numbers of PSA-NCAM-positive granule cells correlated well with the increase in the immunopositive neurites and synaptic terminals on the mossy fiber trajectory. The increase in these PSA-NCAM-immunopositive structures is thought to reflect the enhancement of sprouting and synaptogenesis of mossy fibers by a subset of granule cells newly generated during amygdaloid-kindling and suggests that the reorganization of the mossy fiber system on the normal trajectory at least in part contributes to the acquisition and maintenance of an epileptogenic state. PMID- 14741394 TI - SP600125, a new JNK inhibitor, protects dopaminergic neurons in the MPTP model of Parkinson's disease. AB - Increasing evidence suggests that c-Jun N-terminal kinase (JNK) is an important kinase mediating neuronal apoptosis in Parkinson's disease (PD) model induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). In order to study roles of JNK activity in neuronal apoptosis in this model, we blocked JNK activity in vivo using a specific inhibitor of JNK, SP600125. Our data showed that MPTP-induced phospho-c-Jun of substantial nigral neurons, caused apoptosis of dopaminergic neurons, and decreased the dopamine level in striatal area. We found that inhibiting JNK with SP600125 reduced the levels of c-Jun phosphorylation, protected dopaminergic neurons from apoptosis, and partly restored the level of dopamine in MPTP-induced PD in C57BL/6N mice. These results indicate that JNK pathway is the major mediator of the neurotoxic effects of MPTP in vivo and inhibiting JNK activity may represent a new and effective strategy to treat PD. PMID- 14741395 TI - Is state-dependent alternation of slow dynamics in central single neurons during sleep present in the rat ventroposterior thalamic nucleus? AB - Based upon our previous results in cats, we hypothesized that neurons in the central processor systems of the brain generally exhibit state-dependent dynamics alternation of slow fluctuations in spontaneous activity during sleep. To test the validity of this hypothesis across species, we recorded single neuronal activity during sleep from the ventroposterior (VP) thalamic nucleus in unanesthetized, head-restrained rats. Spectral analysis was performed on successive spike-counts of neuronal activity recorded during three stages of the sleep-wakefulness cycle: wakefulness (W, n=6), slow-wave sleep (SWS, n=20), and paradoxical sleep (PS, n=32). We found that firing of VP neurons displayed white noise-like dynamics over the range of 0.04-1.0 Hz during SWS and 1/f-noise-like dynamics over the same range during PS. We also demonstrated for the first time that the slow dynamics of neuronal activity during quiet wakefulness (but not drowsiness) are white-noise-like. These results suggest that our hypothesis is true across species. During W and SWS, the brain may be considered as under global inhibition. Conversely, PS may represent a state of global disinhibition in the brain, where neuronal activity exhibits 1/f-noise-like dynamics. Fluctuations observed in living organisms may be involved in essential processes in generation and function of sleep states. PMID- 14741396 TI - Experience-dependent changes in intracellular Cl- regulation in developing auditory neurons. AB - A developmental change in GABA and glycine responses, from a depolarization to a hyperpolarization, have been reported for a range of CNS neurons, and has been demonstrated to be due to a developmental decrease in the intracellular Cl- concentration ([Cl-](i)). We examined [Cl-](i) in isolated rat lateral superior olive (LSO) neurons using patch-clamp recordings of glycine gated Cl- currents and by measuring intracellular Cl- -fluorescence. In neurons from 14-16-day-old rats (P14-P16), which had previously received unilateral or bilateral cochlear ablations before the onset of hearing, there was no developmental decrease in [Cl ](i). No significant differences in [Cl-](i) were observed amongst rats with either ipsi- and contralateral ablations. Implanted strychnine pellets also prevented the decrease in [Cl-](i) in most neurons. In some of these neurons in which [Cl-](i) remained high, there was a lack of expression of the K+-Cl- cotransporter 2 (KCC2) mRNA. These results demonstrate that the developmental decrease in [Cl-](i) in LSO neurons is dependent on neuronal activity and that both GABAergic/glycinergic and glutamatergic afferent activity contribute to this maturation of the Cl- regulatory mechanisms. PMID- 14741397 TI - Modulation of sensory-CNS-motor circuits by serotonin, octopamine, and dopamine in semi-intact Drosophila larva. AB - We have introduced an in-situ preparation to induce motor unit activity by stimulating a sensory-CNS circuit, using the third instar larvae of Drosophila melanogaster. Discrete identifiable motor units that are well defined in anatomic and physiologic function can be recruited selectively and driven depending on the sensory stimulus intensity, duration, and frequency. Since the peripheral nervous system is bilaterally symmetric to coordinate bilateral symmetric segmental musculature patterns, fictive forms of locomotion are able to be induced. Monitoring the excitatory postsynaptic potentials (EPSP) on the prominent ventral longitudinal body wall muscles, such as m6 and m12, provides additional insight into how the selective motor units might be recruited within intact animals. We also introduce the actions of the neuromodulators (serotonin, octopamine (OA) and dopamine (DA)) on the inducible patterns of activity within the sensory-motor circuit. The powerful genetic manipulation in Drosophila opens many avenues for further investigations into the circuitry and cellular aspects of pattern generation and developmental issues of circuitry formation and maintenance in the model organism. PMID- 14741398 TI - Nicotinic stimulation modulates tyrosine hydroxylase mRNA half-life and protein binding to the 3'UTR in a manner that requires transcription. AB - Tyrosine hydroxylase (TH) expression increases in adrenal chromaffin cells treated with the nicotinic agonist, dimethylphenylpiperazinium (DMPP; 1 microM). We are using this response as a model of the changes in TH level that occur during increased cholinergic neural activity. Here we report a 4-fold increase in TH mRNA half-life in DMPP-treated cells chromaffin cells that is apparent when using a pulse-chase analysis to measure TH mRNA half-life. No increase is apparent using actinomycin D to measure half-life, indicating a requirement for ongoing transcription. Characterization of protein binding to the TH 3'UTR responsible for stabilization using labeled TH 3'UTR probes and electro-mobility shift assays shows the presence of two complexes both of which are increased by DMPP-treatment. The faster migrating complex (FMC) increases 2.5-fold and the slower migrating complex (SMC) increases 1.5-fold. Both changes are prevented by actinomycin D. Characterization of the protein binding to the TH UTR probes indicates SMC is disrupted by polyribonucleotides, poly (A) and poly (U), while binding to FMC is reduced by poly (CU). Separation of UV crosslinked RNA-protein complexes on SDS polyacrylamide gels shows FMC to contain a single protein whereas SMC contains three proteins. Northwesterns yielded similar results. Comparison of DMPP-induced protein binding with the poly C binding protein (PCBP) involved in hypoxia induced rat PC12 TH mRNA stability indicates none of the bovine UTR binding proteins are the PCBP. Thus, nicotinic stimulation produces a transcription-dependent increase in TH mRNA half-life that is mediated by previously unrecognized TH mRNA binding proteins. PMID- 14741400 TI - Up-regulation and altered distribution of lysyl oxidase in the central nervous system of mutant SOD1 transgenic mouse model of amyotrophic lateral sclerosis. AB - Mutations of the copper-zinc superoxide dismutase (SOD1) gene can result in the development of amyotrophic lateral sclerosis (ALS). The exact cellular mechanisms causing ALS are not known, but oxidative stress is thought to play a prominent role. Lysyl oxidase (LOX) is one of the genes that are known to be up-regulated in ALS patients. In this study, we examined LOX localization in wild type rat and mouse brain sections using immunohistochemistry coupled with laser-scanning confocal microscope. The results showed that LOX, an extracellular matrix protein, was expressed in the choroid plexus, blood vessel walls, brain matrix, and neurons of normal rat and mice. In neurons, LOX was localized within the cytoplasm. LOX immunoreactivity increased in neurons of the spinal cord, brain stem and cortex, and the Purkinje cells of the cerebellum in transgenic G93A SOD1 (mSOD1) mouse model of ALS. In situ hybridization indicated that LOX gene expression was enhanced in the neurons of the spinal cord, brain stem, cortex, caudoputamen and cerebellum in mSOD1 mice compared with wild type controls. LOX enzyme activity was increased in mSOD1 mice. An increase in the amount of LOX mRNA, protein and enzyme activity was coincidental with late stage ALS, indicating that LOX may be associated with the progression of the neurodegenerative process in the mSOD1 model of ALS. PMID- 14741399 TI - Functional differences between two DCLK splice variants. AB - Recently, we have cloned two splice variants of the doublecortin-like kinase (DCLK) gene, called DCLK-short-A and -B, both of which encode calcium/calmodulin dependent protein kinase (CaMK)-like proteins with different C-terminal ends. Using in situ hybridization, we have found that both are highly expressed in limbic structures of the brain and that their expression differs in a number of brain areas. DCLK-short-A is relatively more strongly expressed than DCLK-short-B in the subependymal zone. The DCLK-short-B variant shows stronger expression in the cortex, the ventromedial and dorsomedial hypothalamic nuclei, the arcuate nucleus, the zona incerta and the subincertal nucleus. Also, within the hippocampus, the relative distribution of these two splice variants differs. DCLK short-B expression compared to DCLK-short-A is highest in the CA1 area. The expression of the A variant is highest in the CA3/CA4 area. Additionally, DCLK short-B is expressed at a higher level than DCLK-short-A in the substantia nigra and the mammillary nucleus. Both DCLK-short-A and -B were located in the cytoplasm, however DCLK-short-B was also found specifically in growth cone like structures and near the nucleus. Both DCLK-short proteins phosphorylate autocamtide and syntide, two highly specific CaMK substrates. Finally, removal of the C-terminal end of DCLK-short leads to a 10-fold increase of kinase activity, indicating that the different C-termini represent auto-inhibitory domains. Our results indicate that DCLK-short-A and -B control different neuronal processes that overlap with those controlled by CaMKs. PMID- 14741401 TI - High affinity binding of the Translin/Trax complex to RNA does not require the presence of Y or H elements. AB - Translin and its partner protein, Trax, are components of an RNA binding complex that has been implicated in suppressing translation of several mRNAs by binding to Y and H cis elements contained in these transcripts. However, it is unclear which features of these elements are critical for conferring high affinity binding to the Translin/Trax complex, information that might be useful in identifying other candidate transcripts targeted by this complex. To help clarify this issue, we have assessed the effect of truncating or mutating a segment of the 3'UTR of the protamine-2 transcript which contains both Y and H elements and binds to this complex with high affinity. Our results indicate that high affinity binding to this segment is preserved following extensive mutation of the Y and H elements as long as clusters of G residues are retained. Thus, our findings indicate that the Translin/Trax complex recognizes clusters of G residues rather than RNA sequences that closely match the primary sequence of the Y and H elements. This revised view of the cis elements recognized by the Translin/Trax complex may be useful in future studies aimed at identifying endogenous RNA species targeted by this complex. PMID- 14741402 TI - Ethanol withdrawal-induced up-regulation of the alpha2 subunit of the GABAA receptor and its prevention by diazepam or gamma-hydroxybutyric acid. AB - The gamma-aminobutyric acid type A (GABA(A)) receptor is an important pharmacological target of ethanol. The effect of ethanol withdrawal on the expression of the alpha(2) subunit of this receptor was examined with rat cerebellar granule cells in primary culture. Long-term exposure of these cells to ethanol (100 mM, 5 days) did not affect the abundance of the mRNA for the alpha(2) subunit, as revealed by an RNase protection assay. In contrast, subsequent ethanol withdrawal for 3 h induced a marked increase in the amount of this mRNA (2.6-fold) as well as in that of the encoded polypeptide (2.2-fold), the latter revealed by immunoblot analysis. Exposure of the cells to gamma hydroxybutyric acid (100 mM) during ethanol withdrawal prevented the increase in the amounts of both the alpha(2) mRNA and polypeptide, whereas similar treatment with diazepam (10 microM) blocked the increase in the abundance of the alpha(2) polypeptide but not that in the amount of the alpha(2) mRNA. The effect of gamma hydroxybutyric acid was not blocked by the competitive GABA(B) receptor antagonist SCH 50911(10 microM). Given that the alpha(2) subunit of the GABA(A) receptor mediates the anxiolytic action of benzodiazepines, its up-regulation during discontinuation of long-term ethanol exposure might be relevant to the therapeutic efficacy of these drugs in the treatment of anxiety associated with ethanol withdrawal. PMID- 14741403 TI - The involvement of VEGF receptors and MAPK in the cannabinoid potentiation of Ca2+ flux into N18TG2 neuroblastoma cells. AB - In addition to their inhibitory effects, cannabinoids also exert stimulatory activity which can be detected at the cellular level. In a previous study, we demonstrated a stimulatory effect of the synthetic cannabinoid receptor agonist desacetyllevonantradol (DALN) on Ca(2+) flux into N18TG2 neuroblastoma cells, and suggested a dual mechanism: one pathway mediated by PKA and the other one by protein kinase C (PKC). Here we studied the PKC-mediated effect of DALN on Ca(2+) influx. The stimulatory effect of DALN on Ca(2+) influx was partially blocked by the PKC inhibitor chelerythrine, by the metalloprotease inhibitor o phenanthroline and by the MEK (mitogen-activated protein-kinase kinase, MAPK kinase) inhibitor PD98059. Immunobloting of ERK1/2 MAPK demonstrated phosphorylation by DALN, and indicated the involvement of vascular endothelial growth factor (VEGF) receptor tyrosin kinases (RTKs) in MAPK activation as it was blocked by oxindole-1. Transactivation of the VEGFR-MAPK cascade by DALN involved CB1 cannabinoid receptors coupled to Gi/Go GTP-binding proteins as it was blocked by SR141716A and by pertussis toxin (PTX). The pharmacological implications of this novel mechanism of cannabinoid activity are discussed. PMID- 14741404 TI - Survival of motor neuron gene downregulation by RNAi: towards a cell culture model of spinal muscular atrophy. AB - Gene silencing with double-stranded RNA (RNAi) has proved useful for gene function studies, and should be especially well suited to studying diseases resulting in embryonal lethality where transgenic animal models are difficult to generate. We are applying this approach to the autosomal recessive disease spinal muscular atrophy (SMA). SMA is caused by mutations in the survival of motor neuron gene (SMN). The SMN protein is ubiquitously expressed and plays a role in RNA processing and its reduction in SMA ultimately leads to motor neuron degeneration in the spinal cord. The reasons for this motor neuron selectivity, however, are still unclear. SMN is essential for the viability of most eukaryotic organisms and this has made the generation of animal models of SMA extremely difficult. Here we describe a different approach to study SMN function using RNAi to silence SMN expression in cells. We designed double-stranded small interfering RNA (siRNA) targeted against murine Smn and transfected the murine embryonal terato-carcinoma cell line P19. The siRNAs reduced both Smn RNA and protein levels in the P19 cells compared to controls. These results illustrate that double-stranded RNA can be an effective gene silencing approach even in a protein that is essential for survival and highly expressed, and it could therefore be a valuable tool to study SMN function. PMID- 14741405 TI - Cooperative dimerization of the POU domain protein Brn-2 on a new motif activates the neuronal promoter of the human aromatic L-amino acid decarboxylase gene. AB - The neuronal promoter of the human aromatic L-amino acid decarboxylase (AADC) gene contains a perfectly palindromic element (TB) that conforms to the structure of a POU domain protein binding site of the MORE+2 type. The TB motif (located at nts -900/-872 relative to the neuronal cap site) bears striking similarities with the dimeric Pit-1 binding site from growth hormone gene promoter (GH-1), and it enhanced the activity of the minimal tk promoter in transfected SK-N-BE neuroblastoma cells. In transfected COS-7 cells, the expression of a 3xTB-tk-luc was stimulated up to 11-fold by the overexpressed Brn-2 protein. In AADC gene neuronal promoter, we previously characterized a bipartite regulatory element (ONF for octamer-like/NF-Y, nts -86/-57) that binds Brn-2 and NF-Y proteins in a cooperative manner. We now show that both TB and ONF sites participate in the activation of the neuronal promoter by Brn-2. EMSA experiments showed that the recombinant Brn-2 POU domain dimerized on the TB element in a cooperative manner. By site directed mutagenesis of the POU domain of Brn-2, the dimerization interface on the TB element was localized to the hydrophobic pocket of the POU specific domain and the C-terminal part of the POU homeodomain. PMID- 14741406 TI - Daily rhythm and regulation of clock gene expression in the rat pineal gland. AB - Rhythms in pineal melatonin synthesis are controlled by the biological clock located in the suprachiasmatic nuclei. The endogenous clock oscillations rely upon genetic mechanisms involving clock genes coding for transcription factors working in negative and positive feedback loops. Most of these clock genes are expressed rhythmically in other tissues. Because of the peculiar role of the pineal gland in the photoneuroendocrine axis regulating biological rhythms, we studied whether clock genes are expressed in the rat pineal gland and how their expression is regulated.Per1, Per3, Cry2 and Cry1 clock genes are expressed in the pineal gland and their transcription is increased during the night. Analysis of the regulation of these pineal clock genes indicates that they may be categorized into two groups. Expression of Per1 and Cry2 genes shows the following features: (1) the 24 h rhythm persists, although damped, in constant darkness; (2) the nocturnal increase is abolished following light exposure or injection with a beta-adrenergic antagonist; and (3) the expression during daytime is stimulated by an injection with a beta-adrenergic agonist. In contrast, Per3 and Cry1 day and night mRNA levels are not responsive to adrenergic ligands (as previously reported for Per2) and daily expression of Per3 and Cry1 appears strongly damped or abolished in constant darkness. These data show that the expression of Per1 and Cry2 in the rat pineal gland is regulated by the clock-driven changes in norepinephrine, in a similar manner to the melatonin rhythm-generating enzyme arylalkylamine N-acetyltransferase. The expression of Per3 and Cry1 displays a daily rhythm not regulated by norepinephrine, suggesting the involvement of another day/night regulated transmitter(s). PMID- 14741407 TI - Relaxation of glycine receptor and onconeural gene transcription control in NRSF deficient small cell lung cancer cell lines. AB - Negative regulation of many neuronal genes is mediated by the neuron-restrictive silencer factor (NRSF/repressor element-1 binding transcription factor, REST), which binds to the neuron-restrictive silencer element (NRSE/repressor element-1, RE-1) and thereby represses transcription of neuronal genes in non-neuronal cells. Sequence analysis of 5'-flanking regions of glycine receptor (GlyR) subunit genes revealed a consensus motif for NRSE in the GLRA1 and GLRA3, but not in GLRB, genes. In this study, we examined tumor cell lines for the expression of NRSF, GlyR subunits and onconeural genes. We identified two small cell lung cancer (SCLC) cell lines lacking full-length NRSF/REST as well as its neuronal splice variants. Presence or absence of NRSF as well as its functionality in different SCLC cell lines was additionally shown in reporter gene assays. As GlyR alpha1 is selectively transcribed in NRSF/REST free cells, GlyR alpha1 transcripts might serve as positive signals for NRSF deficient cells. In contrast, GlyR beta is nearly ubiquitously transcribed in the cell lines analyzed and, therefore, should represent a useful marker for neoplastic cells. Sequence analysis of GlyR beta transcripts led to the identification of a new splice variant lacking exon 8, GlyR beta Delta8. This suggests that the lack of NRSF in SCLC cells, resulting in the relaxation of neuronal gene suppression, is an important mechanism underlying paraneoplastic expression. PMID- 14741408 TI - Time-course study of SCG10 mRNA levels associated with LTP induction and maintenance in the rat Schaffer-CA1 pathway in vivo. AB - The maintenance of long-term potentiation (LTP) depends on altered gene expression. Previously, we found the expression of neuronal growth associated protein SCG10, which is involved in neurite outgrowth and neural regeneration, was up-regulated by LTP induction in the rat hippocampal Schaffer-collateral CA1 pathway. Here we studied the temporal expression pattern of SCG10 mRNA after LTP induction using permanently implanted electrodes in the same CA1 pathway. The real-time RT-PCR showed that both SCG10 mRNA 1 and 2 kb forms were increased at the 3 h, but not at 1 or 24 h. In situ hybridization revealed an increase of SCG10 2 kb mRNA level in ipsilateral CA3 and CA1 areas, but not their contralateral counterparts or either side of dentate gyrus. These results suggest that SCG10 may play a role in the maintenance of synaptic plasticity through a transient regulation of microtubule dynamics, which facilitates the structural remodeling of the presynaptic element during the consolidation period. PMID- 14741409 TI - Adrenalectomy further suppresses the NT-3 mRNA response to traumatic brain injury but this effect is not reversed with corticosterone. AB - Fluid percussion injury (FPI) and in situ hybridisation were used to evaluate the expression of NT-3 mRNA in the hippocampus after traumatic brain injury (TBI) in adrenal-intact and adrenalectomised rats (with or without corticosterone replacement). FPI and adrenalectomy independently significantly reduced the expression of NT-3 mRNA in the dentate gyrus (DG) and CA2 region. The effects of adrenalectomy in the CA2 region were partially reversed with corticosterone. In adrenalectomised animals undergoing FPI, a further significant decrease in NT-3 mRNA was observed in the DG, but this was not reversed by corticosterone. Glucocorticoids may, therefore, play a role in the basal regulation of NT-3 in the hippocampus, but the role of glucocorticoids in the modulation of the NT-3 response to TBI is unclear. PMID- 14741410 TI - Adenosine A2A receptor mRNA expression is increased in rat striatum and nucleus accumbens after memantine administration. AB - Quantitative in situ hybridization revealed a significant increase (23-43%) of A(2A) receptor mRNA levels 8-48 h after administration of memantine (25 mg/kg, i.p.) in the striatum and nucleus accumbens regions of rat brain. These results indicate an effect on adenosine A(2A) receptors at the molecular level by a NMDA receptor antagonist. PMID- 14741411 TI - Nogo 3'-untranslated region CAA insertion: failure to replicate association with schizophrenia and demonstration of marked population difference in frequency of the insertion. AB - Novak et al. [Brain Res. Mol. Brain Res. 107 (2002) 183] reported that a CAA insertion in the 3'-untranslated region of the Nogo gene was associated with schizophrenia. We examined the frequency of this CAA insertion in 57 European American subjects with schizophrenia and 243 controls, and in a smaller group of African American subjects (N=72; 20 with schizophrenia). We found a similar frequency of the CAA insertion for patients and controls in both populations, but a large difference in CAA insertion frequency between the two racial groups. PMID- 14741412 TI - Possible association between Cys311Ser polymorphism of paraoxonase 2 gene and late-onset Alzheimer's disease in Chinese. AB - The common polymorphism at codon 311 (C311S) of paraoxonase 2 gene (PON2) was investigated in 165 patients with sporadic late-onset Alzheimer's disease (LOAD) and 174 controls in Chinese. The PON2*C allele frequency was significantly increased in the patients as compared with controls. However, no significant difference was observed after stratification of apolipoprotein E (ApoE) epsilon4 allele. These results suggested that the PON2 polymorphism might be a risk factor for LOAD independent of ApoE epsilon4 status in Chinese. PMID- 14741416 TI - Opioid receptor expression and function. PMID- 14741413 TI - Alterations in the expression of lipid and mechano-gated two-pore domain potassium channel genes in rat brain following chronic cerebral ischemia. AB - To determine the gene expression level of the lipid-sensitive mechano-gated 2P domain K(+) channels in rat cortex and hippocampus after chronic cerebral ischemia, TREK-1, TREK-2 and TRAAK were detected in rats at 3 and 30 days after permanent bilateral carotid artery ligation (BCAL). By using reverse transcription-polymerase chain reaction, we found that TREK-1 and TREK-2 mRNA had no obvious changes in cortex after ischemia, while TRAAK mRNA was significantly increased by 70% after 30 days. In hippocampus, the gene expressions of TREK-1 and TRAAK were increased markedly at 3 days (97% and 87%, respectively) and 30 days (63% and 47%, respectively) after ischemia. However, TREK-2 gene expression level had no change. In-situ hybridization results also showed that TREK-1 gene expressing-positive neurons were increased significantly in cortex and hippocampus after permanent BCAL for 3 and 30 days. These molecular studies provide evidence for an involvement of the lipid-sensitive mechano-gated 2P domain K(+) channels in the BCAL model, which might have neuroprotective effects in cerebral ischemia. PMID- 14741417 TI - Do drugs of abuse impact on HIV disease? AB - Drug abuse, certain lifestyles, access and adherence to drug abuse treatment, and medical consequences of drug abuse remain as important factors that impact on HIV disease among AIDS patients worldwide. Most in vitro and in vivo studies show a significant impact on HIV disease. Epidemiological studies in the past have failed to support these observations. However, new and limited evidence shows that drug abuse may accelerate HIV disease in humans. Research is needed to design new or refine known techniques that more closely mimic natural conditions of HIV disease, and perform additional assessments of basic laboratory, clinical, and epidemiological data to determine whether drug abuse significantly impacts on HIV disease. PMID- 14741418 TI - Drugs of abuse and HIV--a perspective. AB - HIV and drugs of abuse have each been implicated as promoters of human organ specific diseases which include the heart, kidney, lung, CNS etc. Whether the drugs of abuse directly and/or indirectly increase the susceptibility of HIV-1 induced pathology and whether the drugs of abuse and HIV effects are cumulative and/or synergistic remain to be established. The scope of the studies that are needed to address this issue and the limitations inherent in such studies are summarized below. PMID- 14741419 TI - Interaction between HIV and intravenous heroin abuse? AB - In this article I will review the findings of the potential interactive effects of HIV infection and heroin use as observed in both clinical and laboratory studies. Some of the findings are discrepant and I will endeavour to propose models that could accommodate these variant results as well as suggest factors that could be considered in analysing future investigations in order to resolve these apparent discrepancies. PMID- 14741420 TI - Methamphetamine and AIDS: 1HMRS studies in a feline model of human disease. AB - Potential interactions between psychostimulant drugs and infection with feline immunodeficiency virus (FIV) on brain metabolism were evaluated. Four groups of cats were studied: control, FIV positive, methamphetamine (MA) exposed, and FIV positive plus MA exposed. Frontal gray matter, frontal white matter, and caudate brain extracts were studied with proton magnetic resonance spectroscopy (1HMRS). In the frontal white matter, FIV-infected cats showed decreases in creatine and choline, while MA-treated cats had elevated gamma-aminobutyric acid (GABA). The decreased glutamate in FIV cats normalized with MA exposure. FIV and MA both affect brain metabolites individually and combined. 1HMRS is useful for evaluating the effects of FIV and drug abuse in the brain. PMID- 14741421 TI - Association of drug abuse with inhibition of HIV-1 immune responses: studies with long-term of HIV-1 non-progressors. AB - Recreational drug use has been proposed to affect the course of human immunodeficiency virus (HIV) infections. To investigate the effects of substance abuse on HIV infections, we compared virus-specific cytotoxic T lymphocyte (CTL) responses and the expression of IL-16, TGF-beta1, and CXCR4 in three different cohorts of HIV-infected patients: (1) long-term nonprogressors (LT-NPs) of HIV infection who do not use recreational drugs; (2) nondrugs using normal progressors (NPs), and (3) drugs using NPs. Our results show that LT-NPs manifest increased CTL activity and IL-16 expression and decreased expression of TGF-beta1 and CXCR4 compared to NPs, regardless of recreational drug usage. Furthermore, drugs using NPs showed significantly lower levels of CTL and IL-16 expression and increased TGF-beta1 and CXCR4 expression compared to nondrugs using NPs. Our results suggest that recreational drug use may reduce CTL and IL-16 expression and increase the expression of TGF-beta1 and CXCR4, all of which may facilitate progression of HIV infections. PMID- 14741422 TI - Evidence implicating cocaine as a possible risk factor for HIV infection. AB - Epidemiological evidence regarding the impact of drugs of abuse on HIV disease is mixed but points to a likely association between cocaine use (by different routes) and acquisition of either HIV infection or an AIDS-defining illness. In vitro studies indicate that cocaine increases replication of HIV in both stimulated and unstimulated human peripheral blood monocytes. Recent studies using the huPBL SCID mouse model demonstrate that systemic exposure to cocaine enhances HIV replication and spread in vivo. The huPBL SCID mouse and SCID-hu mouse provide potentially useful models for evaluating the impact of other drugs of abuse on HIV infection and the mechanisms mediating these effects. PMID- 14741423 TI - Multiple ways that drug abuse might influence AIDS progression: clues from a monkey model. AB - Whether opiates and other drugs of abuse affect AIDS progression has been an unresolved issue for two decades. Credible evidence has suggested that opiates may exacerbate, retard or have 'no effect' on progression of AIDS. Differences may exist in AIDS-progression outcomes after opiate exposures that relate to neural versus, strictly, somatic AIDS; but it is also likely that conditional variables inherent to drug dependency and the nature of the infectious agents involved allow for differing outcomes. Data from epidemiological studies, and from in vitro and basic immunological studies regarding opiate effects on AIDS progression must be interpreted in light of the conditionality of opiate effects. Caution is sounded, also, about interpreting common depressive immunological effects of opiates as indicators of influence over AIDS progression. Current evidence from the monkey model of AIDS indicates that opiates can, under certain defined conditions, retard AIDS progression. The clinical relevance of these data remains to be defined. Importantly, such data imply that opioids and the endogenous opioid system may represent therapeutic tools and targets for altering AIDS progression. PMID- 14741424 TI - Pneumococcus virulence factor sialidase: a new direction in neuro-AIDS research? AB - The purpose of this presentation is to invite consideration by the research community of the hypothesis that sialidase, a virulence factor of Streptococcus pneumoniae (pneumococcus) and most other opportunistic co-infectious agents associated with HIV infection, advances progression of HIV infection to neuro AIDS. PMID- 14741425 TI - Proteomic fingerprinting of HIV-1-infected human monocyte-derived macrophages: a preliminary report. AB - Mononuclear phagocytes (MP; blood monocytes, alveolar, lymph node, and brain macrophages and microglia) are vehicles for dissemination and principle target cells for human immunodeficiency virus type 1 (HIV-1) infection. Notably, viral persistence in macrophages occurs despite ongoing phagocytic, intracellular killing, innate and adaptive immune responses. To assess potential pathways for how HIV-1 may bypass antiviral MP responses, we used proteomic tests to evaluate protein fingerprints of HIV-1-infected human monocyte-derived macrophages 7 days after viral infection. By using weak cation exchange chips, 58 proteins were found up- or down-regulated after HIV-1(ADA) infection. Several of these proteins were identified by microsequencing. It is probable that cellular proteins identified by proteomic fingerprinting could assist in unraveling how persistent viral infection occurs in MP lineage cells. Moreover, this evolving technology can be utilized to unravel changes in immune activities initiated by interactions between virus, environmental cues and drugs of abuse. PMID- 14741426 TI - Neurodegenerative disease and the neuroimmune axis (Alzheimer's and Parkinson's disease, and viral infections). AB - The appearance of activated immune cells and the accumulation of inflammation associated proteins are common phenomena associated with neurodegenerative diseases. These inflammatory components of central nervous system (CNS) diseases have most often been described in the context of an immune response to damage and cell loss already occurring in the affected brain area. There has, however, been a renewed interest in how the neuroimmune axis might itself be involved in the etiology of these neurodegenerative diseases, particularly in cases involving slow, chronic, progressive neuropathology. This review addresses immune activation in Alzheimer's, Parkinson's, and brain viral infections that may be causative of, rather than responsive to, the observed neuronal loss in these pathologies. PMID- 14741427 TI - Microglial activation by uptake of fDNA via a scavenger receptor. AB - The fate of the fragmented DNA (fDNA) observed in neuronal nuclei in Alzheimer brain is unknown. However, its fate is suggested as fDNA is found in the cytoplasm of adjacent activated microglia. After a brief incubation with fDNA, approximately 70% of microglia had fDNA in their cytoplasm, were activated, and overexpressed interleukin-1beta. Microglial activation enhanced uptake whereas blocking scavenger receptors suppressed this uptake. These results suggest that the brain rids itself of fDNA from dying neurons through microglial uptake, activation, and overexpression of IL-1. Such overexpression of IL-1 in Alzheimer brain has been linked to Alzheimer pathogenesis. PMID- 14741428 TI - P2X7 receptor modulation of beta-amyloid- and LPS-induced cytokine secretion from human macrophages and microglia. AB - To test whether extracellular ATP can play a role in the neuroimmunopathology of Alzheimer's disease (AD), we evaluated the capacity of the ATP-binding purinoreceptor, P2X7, to modulate cytokine secretion on cultured human macrophages and microglia pre-activated 24 h with the 42 amino acid beta-amyloid peptide (Abeta(1-42)) or lipopolysaccharide. Thirty minutes of exposure to the selective P2X7 agonist 2'-3'-O-(4-benzoylbenzoyl)adenosine 5'-triphosphate (BzATP) resulted in the secretion of IL-1beta after either Abeta(1-42) or LPS stimulation of human macrophages that was dependent on the concentration of the stimulus used to pre-activate the cells. Further tests on human microglia treated with BzATP (300 microM) resulted in a 1.5- and 3.5-fold enhancement of IL-1alpha and IL-1beta secretion, respectively, from cells pre-activated by 10 microM Abeta(1-42) and a 1.6- and 3.9-fold enhancement of IL-1alpha and IL-1beta secretion, respectively, from cells pre-activated by 1 microg/ml LPS. BzATP induction of IL-1alpha and IL-1beta secretion from microglia was completely reversed by pre-incubation of the cells with the P2X7 antagonist, adenosine 5' triphosphate 2',3'-acyclic dialcohol (oxidized ATP). In contrast to its effects on IL-1alpha and IL-1beta secretion, BzATP induced TNF-alpha after LPS stimulation, but not after stimulation with Abeta(1-42), induced IL-18 secretion regardless of whether microglia were pre-activated and attenuated IL-6 secretion after either LPS or Abeta(1-42) pre-activation. These results demonstrate that extracellular ATP can modulate Abeta-induced cytokine secretion from human macrophages and microglia and thus may play a role in the neuroimmunopathology of AD. PMID- 14741429 TI - APOE genotype-specific differences in human and mouse macrophage nitric oxide production. AB - Individuals expressing an APOE4 genotype demonstrate increased Alzheimer's disease (AD) neuropathology and a decreased onset age. The APOE4 gene may act by modulating the CNS immune response. Using human monocyte-derived macrophages (MDM), we show a significantly greater increase in NO production during immune activation in MDM from APOE4 AD patients compared to normal, age-matched individuals or to AD patients with an APOE 3/3 genotype. Microglia and peritoneal macrophages from APOE4 targeted replacement mice demonstrate a similar increase in NO compared to the APOE3 targeted replacement mice. The enhanced macrophage responsiveness and the increased production of NO in APOE4 AD patients may predispose the CNS to an increased potential for nitration and nitrosation, consistent with the redox imbalance and neuroinflammatory state seen in AD. PMID- 14741430 TI - Protective effect of the SOD/catalase mimetic MnTMPyP on inflammation-mediated dopaminergic neurodegeneration in mesencephalic neuronal-glial cultures. AB - Reactive oxygen species (ROS) produced by activated microglia are deleterious to neurons. In this study, we studied the effect of Mn(III)tetrakis(1-methyl-4 pyridyl)porphyrin (MnTMPyP), a superoxide dismutase/catalase mimetic, on the lipopolysaccharide (LPS)-induced degeneration of dopaminergic neurons in rat mesencephalic neuroglia cultures. MnTMPyP exhibited a significantly protective effect against LPS (5 ng/ml)-induced neurotoxicity as determined by [3H]dopamine uptake and immunocytochemical analysis. MnTMPyP significantly attenuated LPS induced production of superoxide free radical and prostaglandin E(2) (PGE(2)) in microglia. These results indicate that MnTMPyP may potentially be used for the treatment of inflammation-related degenerative neurological disorders. PMID- 14741431 TI - The role of cellular immune response in Theiler's virus-induced central nervous system demyelination. AB - Theiler's murine encephalomyelitis virus (TMEV) persists in spinal cord white matter of susceptible mice (e.g., SJL/J), resulting in chronic inflammation and demyelination. Reconstitution of severe combined immunodeficient (SCID) mice with CD4(+) T- or CD8(+) T-lymphocytes results in extensive TMEV-induced demyelination, and depletion of CD8(+) T-lymphocytes in the early or late phase of the disease decreases the extent of demyelination, indicating that the cellular immune response against the virus plays a key role in myelin destruction. In susceptible mice, the demyelinated lesions are characterized by infiltration of a large numbers of B- and T-lymphocytes; whereas in mice resistant to TMEV-induced demyelination (e.g., C57BL/6), virus clearance requires infiltration of between 2.9 x 10(5) and 5.7 x 10(5) CD8(+) T-lymphocytes and between 3.4 x 10(5) and 6.1 x 10(5) CD4(+) T-lymphocytes per mouse in the brain 5 9 days post infection. Transgenic expression of capsid proteins of TMEV abrogates resistance in C56BL/6 mice, rendering the mice susceptible to TMEV persistence and demyelination. Comparison of the kinetics of virus replication and B- and T lymphocyte infiltration in mice lacking key adhesion molecules (L-selectin (L sel(-/-)), P-selectin (P-sel(-/-)), intracellular adhesion molecule-1 (ICAM-1(-/ )), or leukocyte function-associated antigen-1 (LFA-1(-/-))) demonstrates a role for individual adhesion molecules in recruitment of immune cells into central nervous system (CNS), but the role is not significant to prevent eventual virus clearance. PMID- 14741432 TI - Chronic morphine treatment differentiates T helper cells to Th2 effector cells by modulating transcription factors GATA 3 and T-bet. AB - Chronic morphine treatment in animal models has been shown to alter a number of immune parameters including suppression of cellular immunity. T helper cell differentiation into Th2 effector cell may be a major contributing factor to impaired cellular immunity following chronic drug abuse. We had previously shown that chronic morphine treatment in vivo and in vitro decreases IL-2 and IFNgamma (Th1) protein levels and increases IL-4 and IL-5 (Th2) protein levels in a time dependent manner. In addition in this paper, we show that chronic morphine treatment resulted in a decrease in IFNgamma and IL-2 mRNA and an increase in IL 4 and IL-5 mRNA accumulation in murine splenocytes. Furthermore, chronic morphine treatment inhibited IFNgamma promoter activity and increased IL-4 promoter activity in respective promoter transfected primary T cells. In addition, we also demonstrate that chronic morphine treatment resulted in an increase in GATA 3 binding to DNA consensus elements in electromobility shift assays and an increase in GATA 3 protein and mRNA levels. In contrast, chronic morphine treatment resulted in a decrease in T-bet mRNA levels. From these data, we conclude that chronic morphine treatment differentiates T helper cell to Th2 effector cells by modulating key master switches that results in committing T helper cell to a Th2 phenotype. PMID- 14741433 TI - Mechanisms for impaired effector function in alveolar macrophages from marijuana and cocaine smokers. AB - Lung macrophages provide a first line of host defense against inhaled pathogens and their function is impaired in the lungs of inhaled substance abusers. In order to investigate the mechanism for this impairment, alveolar macrophages (AM) were recovered from nonsmokers (NS), regular tobacco smokers (TS), marijuana smokers (MS), or crack cocaine smokers (CS), and evaluated for their production of nitric oxide (NO) and the role of NO as an antimicrobial effector molecule. AM from NS and TS efficiently killed Staphylococcus aureus and their antibacterial activity correlated closely with the production of nitrite and the expression of mRNA encoding for inducible nitric oxide synthase (iNOS). In contrast, AM collected from MS and CS exhibited limited antimicrobial activity that was not affected by an inhibitor of iNOS, or associated with expression of iNOS. Treatment with either granulocyte/macrophage colony-stimulating factor (GM-CSF) or interferon-gamma restored the ability of these cells to produce NO and to kill bacteria. These findings confirm a significant role for NO as an antibacterial effector molecule used by normal human AM and suggest that this host defense mechanism is suppressed by habitual exposure to inhaled marijuana or crack cocaine in vivo. PMID- 14741434 TI - Sympathetic nervous system regulation of immunity. PMID- 14741435 TI - Cannabinoid receptors and T helper cells. AB - We have reported that injection of marijuana cannabinoids, such as Delta(9) tetrahydrocannabinol (THC), into mice, followed by infection with Legionella pneumophila (Lp), suppresses the development of cell-mediated immunity T helper 1 (Th1) activity. These effects are accompanied by suppression of interleukin (IL) 12 and interferon (IFN) gamma production and enhancement of IL-4 production suggesting THC-induced T helper cell biasing. In the current report, other T helper cell biasing mechanisms were studied. Mice were injected with THC followed 18 h later by a challenge infection with Lp. Two-hour post-infection, spleens were removed and analyzed for mRNA to either IL-12Rbeta2 or GATA3 gene products. The results showed that THC suppressed IL-12Rbeta2 but increased GATA3. Receptor antagonists for CB1 (SR141716A, SR1) and CB2 (SR144528, SR2) were also injected to analyze the involvement of cannabinoid receptors. It was determined that SR1 attenuated the THC suppression of IL-12Rbeta2, while SR2 attenuated the increase in GATA3 mRNA. These results suggest that THC suppresses Th1 biasing activity such as IL-12Rbeta2 by a CB1 mediated mechanism and enhances the Th2 biasing activity, GATA3, by a CB2 mechanism. This dichotomy of receptor involvement might result from differential expression and/or signaling function of CB1 and CB2 on Th1 and Th2 cells. PMID- 14741436 TI - Cocaine modulates cytokine and enhances tumor growth through sigma receptors. AB - Sigma receptors are intracellular receptors that interact with a variety of psychotropic ligands, including cocaine. Administration of cocaine to mice promoted the in vivo growth of a syngeneic lung cancer cell line and identical effects were observed with PRE 084, a selective sigma(1) receptor agonist. Increased tumor growth was accompanied by an increase in IL-10 and a decrease in IFN-gamma production in splenocytes and at the tumor site. The tumor-promoting effects produced by both cocaine and PRE 084 were abrogated by administration of specific antibodies to IL-10, or by administration of a sigma(1) receptor antagonist. We conclude that sigma(1) receptor ligands, including cocaine, augment tumor growth via a cytokine-dependent, receptor-mediated mechanism that involves regulation of T helper 1/T helper 2 cytokine balance. PMID- 14741437 TI - RXR-induced TNF-alpha suppression is reversed by morphine in activated U937 cells. AB - Deficiency in vitamin A has been associated with adverse clinical outcomes in drug users with HIV-1 infection. Retinoids have been demonstrated to suppress proinflammatory cytokine production by immune cells in vitro. These effects are induced by ligand-mediated activation of the retinoid receptors--retinoic acid receptor (RAR) and retinoid X receptor (RXR). In these studies, the effects of all-trans-retinoid acid (ATRA, a RAR agonist), 9-cis-retinoic acid (9cis RA; RAR and RXR agonist), LG101305 (RXR agonist), LG100815 (RAR antagonist) and LG101208 (RXR antagonist) on TNF-alpha production by phytohemagglutanin-activated U937 cells and the modulation of these effects by morphine were examined. TNF-alpha production was suppressed in all cultures exposed to retinoid agonist and antagonist agents. For cells exposed to RXR agonists or RAR antagonist, incubation with morphine resulted in the reversal of TNF-alpha suppression and this effect was inhibited by naloxone. These data suggest that interactions between RXR and morphine are involved in the immune effects of retinoids on TNF alpha production by activated U937 cells. Such information may be important for understanding interactions between drugs of abuse and immune function in individuals with chronic proinflammatory states such as HIV-1 infection. PMID- 14741438 TI - Neurological basis of drug dependence and its effects on the immune system. AB - This review summarizes some of the major points discussed by participants in the symposium session on effects of drugs of abuse on both neurologic and immune systems. Speakers in this session are acknowledged experts and biomedical scientists in the rapidly expanding field of studies of abuse drugs on immune responses, especially as related to the effects of increased susceptibility to infections, including opportunistic infections related to AIDS. The important topics specifically discussed in this session included discussion of the neurobiology of addiction in regards to cell biology of the central nervous system and altered physiological and behavioral functions. Using experimental rodent models, description of effects of heroin or cocaine, especially self administration of these drugs, on immune cell deficiency and HPA activation, was reviewed as well as effects on important proinflammatory cytokines like TNFalpha. A model system concerning acute morphine withdrawal on ex vivo immune responses by murine cells was described in detail, including effects of such withdrawal on splenocyte or macrophage responses to bacterial LPS. The chronic exposure of rodents to a drug such as cocaine or morphine was described in terms of CNS neurochemical alterations as related to immune responsiveness. The effects of the legal drug nicotine, now known to be the addictive substance of cigarette smoke, was discussed in regards to effects on both the neurologic and immunologic system in rodents in terms of antibody formation and T cell function, related to HBA activation and proinflammatory cytokine responses. It is apparent from this session, that studies concerning the impact of drugs of abuse on the brain-immune axis and relationship to the immune system constitute a rapidly expanding area and warrant further interest of biomedical scientists. PMID- 14741439 TI - Immune cell activity during the initial stages of withdrawal from chronic exposure to cocaine or morphine. AB - The immunosuppression accompanying illicit drug use has been shown to contribute to a decreased resistance to a variety of pathogens; however, there is relatively little information on how long these effects persist following withdrawal from chronic drug exposure. To begin to address this question, Sprague-Dawley male rats were administered either cocaine (10 mg/kg, i.p., b.i.d.) for 7 days or morphine (escalating doses up to 40 mg/kg, s.c., b.i.d.) for a 10-day period. Control groups of animals received similar saline injections for equivalent time periods. Drug administration was abruptly discontinued and animals were sacrificed at 2, 24, 72 or 96 h following the last dose. At these time points, proliferation responses of peripheral blood T-lymphocytes stimulated by concanavalin A (Con A) and plasma levels of corticosterone were measured. Plasma corticosterone levels of cocaine- or morphine-treated animals were found to be significantly elevated 24 h following drug cessation as compared to saline animals. At this time, proliferation responses were significantly decreased and were further suppressed during cocaine and morphine withdrawal at 96 and 72 h, respectively. These results suggest that abrupt cessation of cocaine or morphine administration leads to activation of stress-related pathways that may contribute to an increased susceptibility of infection during the initial withdrawal phase. PMID- 14741440 TI - Paradoxes of immunosuppression in mouse models of withdrawal. AB - Previously, our laboratory showed that either abrupt (AW) or precipitated withdrawal (PW) from morphine led to profound suppression of murine splenic antibody responses to sheep red blood cells at 24 h post-withdrawal. In the present studies, we examined the immune mechanisms mediating suppression at that time point. A co-culture method was used to examine whether cells from withdrawn mice had (1) a deficit in function and/or (2) contained populations of suppressor cells. To examine the first hypothesis, cells from normal mice were co-cultured with cells from withdrawn mice in a 1:3 ratio (normal/withdrawn). To test the second hypothesis, the ratio was reversed. The results were paradoxical. Co culture of cells in a 1:3 ratio showed that spleen cells from withdrawn mice had a deficit in macrophage function. Spleen cells from withdrawn mice also showed decreased mRNA levels of IL-1beta, IL-1-Ra, and TNF-alpha and a suppression of co stimulatory molecule expression. To examine the second hypothesis, cells were co cultured in a 3:1 ratio (normal/withdrawn). In this paradigm, spleen cells from abrupt withdrawn mice were shown to contain populations of both suppressor macrophages and B-cells. In vivo experiments carried out on mice 24 h post withdrawal showed increased sensitivity to the lethal effects of LPS and increased production of TNF-alpha, implying a state of macrophage activation. Thus evidence for both suppressed and activated macrophages has been obtained in mice 24 h after abrupt withdrawal from morphine. PMID- 14741441 TI - Antibody response and allogeneic mixed lymphocyte reaction in mu-, delta-, and kappa-opioid receptor knockout mice. AB - The implication of opioid receptors in immune response has been studied using mu , delta- and kappa-opioid receptor knockout mice. The mutant animals were compared to their wild-type (WT) counterparts for antibody (Ab) response to the prototype Ag keyhole limpet hemocyanin (KLH). Kappa-receptor deficient mice displayed higher Ab titers for either total Ig, IgM, IgG1 or IgG2a isotypes, whereas mu and delta animals behaved as wild-type mice. Therefore, endogenous kappa-receptor activation would tonically inhibit Ab response. Opioid receptor deficient mice were also used to investigate the immunosuppressive action of naltrindole, a delta-opioid receptor antagonist, shown earlier to inhibit graft rejection and the allogeneic mixed lymphocyte reaction (MLR) in vitro. Naltrindole and two related compounds inhibited MLR performed with lymphocytes from wild-type and delta-opioid receptor knockout mice. These compounds also suppressed MLR assayed with cells from triple mu/delta/kappa-opioid receptor mutants. We therefore demonstrate that naltrindole immunosuppressive activity is not mediated by any of the three mu-, delta- or kappa-opioid receptors, but by a target which remains to be discovered. PMID- 14741442 TI - Cannabinoids and morphine differentially affect HIV-1 expression in CD4(+) lymphocyte and microglial cell cultures. AB - The influence of substances of abuse on the progression of HIV-1 infection is controversial, and pharmacologic factors have been postulated as a potential explanation for conflicting data arising from epidemiological studies and animal models. In the present study, cell culture models of HIV-1 infection were used to test this hypothesis. The synthetic cannabinoid WIN 55,212-2 was found to potently inhibit HIV-1 expression in a concentration- and time-dependent manner in CD4(+) lymphocyte and microglial cell cultures. In sharp contrast, morphine either inhibited or stimulated viral expression, depending upon the time of drug exposure, and marked differences were observed between CD4(+) and microglial cells. Also, WIN 55,212-2 inhibited the stimulatory effect of morphine in HIV-1 infected CD4(+) cells. These in vitro findings support the notion that pharmacologic factors need to be considered in epidemiological studies and animal models that pertain to HIV-1 infection. PMID- 14741443 TI - Cannabinoid-mediated exacerbation of brain infection by opportunistic amebae. AB - Recent reports indicate a higher frequency of brain infections with opportunistic amebae of the genus Acanthamoeba among immune compromised individuals, including AIDS patients. We have demonstrated, using a murine model of Granulomatous Amebic Encephalitis (GAE), that the major psychoactive and immune suppressive component in marijuana delta-9-tetrahydrocannabinol (THC) exacerbates infection by these amebae. Mice administered THC and infected with Acanthamoeba exhibited dose related higher mortalities than infected vehicle controls. The greater severity of disease for THC-treated mice was accompanied by decreased accumulation of macrophage-like cells at focal sites of infection in the brain. Furthermore, THC administration resulted in decreased levels of mRNA for the pro-inflammatory cytokines interleukin-1 alpha, interleukin-1 beta, and tumor necrosis factor alpha for neonatal rat microglia co-cultured with Acanthamoeba. These results indicate a potential for marijuana to alter the capacity of brain macrophage-like cells to mount a full complement of immune responsiveness to brain infection by opportunistic amebae. PMID- 14741444 TI - Morphine inhibits Fc-mediated phagocytosis through mu and delta opioid receptors. PMID- 14741445 TI - Immune, neuroendocrine, and somatic alterations in animal models of human heroin abuse. AB - We investigated immune, endocrine, and somatic alterations using two animal models of human heroin administration. In a heroin self-administration paradigm, we observed changes in immune function which suggest that the cycle of intermittent drug use is actually a stressor, which in turn not only exacerbates craving and drug-seeking behavior but also collaterally causes suppression of immune function and therefore susceptibility to disease. In another model of rats made physically dependent to heroin, we show that immune function is more broadly compromised, leading to evidence of infection, followed by chronic activation of innate immune function, cachexia, and weight loss. PMID- 14741446 TI - Signaling pathway-related gene expression in neuroimmunoregulation. PMID- 14741447 TI - Interleukin-6 in the aging brain. AB - Astrocytes, microglia, and neurons express the cytokine interleukin-6 (IL-6), which in the brain has been suggested to reduce food intake, inhibit memory and learning, cause neurodegeneration, and exacerbate sickness behavior induced by other cytokines. Recent evidence indicates IL-6 levels are increased in brain of healthy aged animals, thus it may play a role in the neurophysiological manifestations of old age. The purpose of this brief report is to discuss the new evidence that suggests an age-related increase in brain IL-6 and the impact this inflammatory cytokine may have on "successful" aging. PMID- 14741565 TI - [18F]-2-fluoro-2-deoxyglucose transport kinetics as a function of extracellular glucose concentration in malignant glioma, fibroblast and macrophage cells in vitro. AB - FDG-PET is used to measure the metabolic rate of glucose. Transport and phosphorylation determine the amount of hexose analog that is phosphorylated and trapped. Competition occurs for both events, such that extracellular glucose concentration affects the FDG image. This study investigated the effect of glucose concentration on the rate of FDG accumulation in three cell lines. The results show that extracellular glucose concentration has a greater impact on the rate of FDG accumulation than the relative abundance of GLUT transporter subtypes. PMID- 14741566 TI - Pharmacokinetics and tumor retention of 125I-labeled RGD peptide are improved by PEGylation. AB - Tumor growth and metastasis are angiogenesis dependent. Overexpression of integrin alphavbeta3 in angiogenic vessels as well as various malignant human tumors suggests the potential of suitably labeled antagonists of this adhesion receptor for radionuclide imaging and therapy of tumors. Small head-to-tail cyclic peptides including the Arg-Gly-Asp (RGD) amino acid sequence have been radiolabeled and studied in preclinical animal models. However, the fast blood clearance, high kidney and liver uptake, and rapid washout from tumors make this type of tracer ineffective for clinical applications. In this study we modified the cyclic pentapeptide c(RGDyK) with monofunctional methoxy-PEG (mPEG, M.W. = 2,000) and labeled the RGD-mPEG conjugate with 125I. We studied the tumor targeting efficacy and in vivo pharmacokinetic properties of 125I-RGD-mPEG by means of direct tissue sampling and autoradiography in mice xenografted subcutaneously with U87MG glioblastoma. Compared to the 125I-RGD analog, this PEGylated RGD peptide revealed faster blood clearance, lower kidney uptake, and prolonged tumor uptake without compromising the receptor targeting ability. PMID- 14741567 TI - Pharmacological properties of hydrophilic and lipophilic derivatives of octreotate. AB - Derivatives of somatostatin (SST) represent the most important peptides for receptor targeting in oncological applications. Whereas the pharmacophor in somatostatin receptor-affine substances has been thoroughly investigated, the influence of modifications at the N-terminal has not yet been systematically studied. In order to investigate the influence of hydrophilic versus lipophilic modifications at the N-terminal end, a series of homologous derivatives of Tyr3 octreotate modified with oligomers of ethylene glycol or fatty acids were synthesized. For this purpose, Tyr3-octreotate was assembled using solid phase peptide synthesis and the fatty acids or oligomers of ethylene glycol were conjugated to the N-terminal end. The oligomers of ethylene glycol were activated by 4-nitrophenylchloroformate to obtain carbamate-linked hydrophilic compounds. The receptor affinities of these compounds were determined by competition experiments with [125I]Tyr3-octreotide on rat cortex membranes. The hydrophilic derivatives and the short chain lipophilic derivatives revealed IC50 values between 0.66 +/- 0.02 nM and 2.16 +/- 0.31 nM respectively. After labeling with (125)I the organ distribution of selected derivatives was investigated in Lewis rats bearing the rat pancreatic tumor CA20948. All of the compounds showed high tumor uptake. The peptides conjugated to oligomers of ethylene glycol showed low uptake into the liver and kidneys. Increasing the length of the fatty acids resulted in a remarkable decrease in kidney uptake. In conclusion, the systematic modifications at the N-terminal result in a low effect on the receptor affinity but allow the modulation of the pharmacokinetic properties of octreotide derivatives. PMID- 14741568 TI - Imaging of adenovirus-mediated expression of human sodium iodide symporter gene by 99mTcO4 scintigraphy in mice. AB - We have evaluated the feasibility of human sodium iodide symporter (hNIS) as a reporter gene in vitro and in vivo. Recombinant adenovirus encoding hNIS (Rad hNIS) was introduced to FRO cell for 48 hours. Western blotting and 99mTcO4 uptake study revealed functional hNIS expression in the cell. Rad-hNIS was injected to BALB/c mice via tail vein. 99mTcO4 gamma scintigraphy, biodistribution study, and RT-PCR analysis demonstrated a preferential hepatic uptake of 99mTcO4, which was observed for up to one week. Thus, hNIS can be utilized as an effective reporter gene for noninvasive/repeated imaging, in combination with 99mTcO4. PMID- 14741569 TI - Pharmacokinetics and biodistribution of [111In]-avidin and [99mTc]-biotin liposomes injected in the pleural space for the targeting of mediastinal nodes. AB - Pharmacokinetics and mediastinal node uptake of [111In]-avidin and [99mTc]-biotin liposomes following either intrapleural (pleural) or intraperitoneal (ip) injection were determined using scintigraphic imaging. Biodistribution results of [111In]-avidin at 44 h showed 3.3% uptake in mediastinal nodes by pleural injection vs 1.3% with ip injection. Mediastinal node accumulation with [99mTc] biotin-liposomes was not different between injections (0.6% ip vs 0.5% pleural). This study demonstrates the potential of the pleural route as a technique for mediastinal node targeting using the avidin/biotin-liposome system. PMID- 14741570 TI - Imaging recognition of multidrug resistance in human breast tumors using 99mTc labeled monocationic agents and a high-resolution stationary SPECT system. AB - Imaging recognition of multidrug-resistance by 99mTc-labeled sestamibi, tetrofosmin and furifosmin in mice bearing human breast tumors was evaluated using a high-resolution SPECT, FASTSPECT. Imaging results showed that the washout rates in drug-resistant MCF7/D40 tumors were significantly greater than that in drug-sensitive MCF7/S tumors. Furifosmin exhibited greater washout from both MCF7/S and MCF7/D40 than sestamibi, while tetrofosmin washout was greater than sestamibi in MCF7/D40 only. Feasibility of the monocationic agents for characterizing MDR expression was well clarified with FASTSPECT imaging. PMID- 14741571 TI - Imaging of gliomas with Cis-4-[18F]fluoro-L-proline. AB - Tumor imaging with cis-4-[18F]fluoro-L-proline (cis-FPro) was compared to that of L-[3H]proline and L-[3H]methionine in F98 rat gliomas by dual-tracer autoradiography. All tracers exhibited high accumulation in the tumors but in the normal brain significant uptake was observed for L-[3H]methionine only. Tumor extent on autoradiograms with L-[3H]proline and L-[3H]methionine was identical to that of histological staining while autoradiograms of cis-FPro showed diffuse uptake in the penumbra of some tumors. First PET studies in 7 patients with cerebral gliomas demonstrated accumulation of cis-FPro in tumor areas with enhancement of Gd-DTPA on MR scans. Uptake of cis-FPro in normal brain tissue was negligible. In one patient with a glioblastoma accumulation of cis-FPro was also found in two brain areas without enhancement of Gd-DTPA on MR scans. Control of MRI suggested tumor growth in these areas at further follow up. Our results indicate that in most gliomas increased cis-FPro uptake is restricted to areas with disruption of the BBB which limits its clinical utility. PMID- 14741572 TI - Comparative studies of potential cancer biomarkers carbon-11 labeled MMP inhibitors (S)-2-(4'-[11C]methoxybiphenyl-4-sulfonylamino)-3-methylbutyric acid and N-hydroxy-(R)-2-[[(4'-[11C]methoxyphenyl)sulfonyl]benzylamino]-3 methylbutanamide. AB - (S)-2-(4'-[11C]methoxybiphenyl-4-sulfonylamino)-3-methylbutyric acid ([11C]MSMA) and N-hydroxy-(R)-2-[[(4'-[11C]methoxyphenyl)sulfonyl]benzylamino]-3 methylbutanamide ([11C]CGS 25966), carbon-11 labeled matrix metalloproteinase (MMP) inhibitors, have been synthesized for evaluation as new potential positron emission tomography (PET) cancer biomarkers. [11C]MSMA was prepared by appropriate precursor (S)-2-(4'-hydroxybiphenyl-4-sulfonylamino)-3-methylbutyric acid tert-butyl ester, which was synthesized in eight steps from amino acid (L) valine in 39.4% chemical yield. This precursor was labeled by [11C]methyl triflate through O-[11C]methylation method at the hydroxyl position of biphenol under basic conditions, followed by a quick acid hydrolysis and isolated by solid phase extraction (SPE) purification to produce pure target compound [11C]MSMA in 35-55% radiochemical yield, based on 11CO2, decay corrected to end of bombardment (EOB), and 20-25 min synthesis time. [11C]CGS 25966 was prepared in our previous work starting from amino acid (D)-valine. The biodistribution of [11C]MSMA and [11C]CGS 25966 were determined at 45 min post iv injection in breast cancer animal models MCF-7's transfected with IL-1alpha implanted athymic mice and MDA MB-435 implanted athymic mice. The results showed the uptakes of [11C]MSMA and [11C]CGS 25966 in these tumors were 0.95 and 0.42%dose/g in MCF-7's transfected with IL-1alpha implanted mice, 0.98 and 1.53%dose/g in MDA-MB-435 implanted mice, respectively; the ratios of tumor/muscle (T/M) and tumor/blood (T/B) were 1.21 and 1.09 (T/M, MCF-7's), 0.99 and 0.84 (T/B, MCF-7's), 1.38 and 1.27 (T/M, MDA-MB 435), 1.27 and 1.95 (T/B, MDA-MB-435), respectively. The micro-PET images of [11C]MSMA and [11C]CGS 25966 in both breast cancer athymic mice were acquired for 15 min from a MCF-7's transfected with IL-1alpha and/or MDA-MB-435 implanted mouse at 45 min post iv injection of 1 mCi of the tracer using a dedicated high resolution (<3 mm full-width at half-maximum) small FOV (field-of-view) PET imaging system, Indy-PET II scanner, developed in our laboratory, which showed both tumors were invisible with both tracers. The results were compared. From our results, we concluded that both [11C]MSMA and [11C]CGS 25966 might be unsuitable as PET tracers for cancer imaging. PMID- 14741573 TI - 99mTc labelled model drug carriers - labeling, stability and organ distribution in rats. AB - The surface characteristics of intravenously administered particulate drug carriers decisively influence the protein adsorption that is regarded as a key factor for the in vivo fate of the carriers. We labeled surface-modified polymer particles with the gamma-emitting radioisotope 99mTc in order to test their properties in blood and follow their in vivo fate. The biodistribution was different in various types of polymer particles. As expected, labeled particles were found in the mononuclear phagocyte system in a large scale but markedly different biodistribution for some particles were also shown. PMID- 14741574 TI - Imaging of intestinal lymphocyte homing by means of pinhole SPECT in a TNBS colitis mouse model. AB - BACKGROUND AND AIMS: The increasing knowledge of the molecular basis of leukocyte trafficking results in the development of novel anti-inflammatory strategies for inflammatory bowel disease (IBD). For optimal evaluation of therapy efficacy, information about inflammatory activity in bowel segments or lymphocyte recirculation and kinetics in the follow-up of experimental treatment for IBD is needed. The aim of this study was to evaluate a non-invasive scintigraphic technique, able to assess lymphocyte trafficking in a trinitrobenzene sulfonic acid (TNBS) induced mouse colitis model of IBD. METHODS: TNBS sensitized and non sensitized murine total splenocytes were labeled in vitro with 111In-oxine and injected into either control or TNBS colitis BALB/c mice. Biodistribution and specific radioactive uptake, representing transferred cells, were determined by serial dedicated animal planar scintigraphy and pinhole SPECT of the abdomen 4, 24 and 48h post injection of labeled cells. In addition, the severity of inflammation was determined by histological scoring. RESULTS: Migration of 111In labeled splenocytes to the colon increased in time and was maximal at 48h after administration. The highest specific radioactive uptake ratio in the colon after 48h was observed in mice with TNBS colitis that received TNBS sensitized lymphocytes. Histological scoring confirmed the presence of colitis in the TNBS treated groups. CONCLUSION: Homing of TNBS-sensitized lymphocytes can be assessed in vivo by means of dedicated animal pinhole SPECT. Generally, this technique enables serial measurement of specific cell trafficking with potential of in vivo evaluation of novel anti-inflammatory strategies in inflammatory bowel disease. PMID- 14741575 TI - A novel electrophilic synthesis and evaluation of medium specific radioactivity (1R,2S)-4-[18F]fluorometaraminol, a tracer for the assessment of cardiac sympathetic nerve integrity with PET. AB - (1R,2S)-4-[18F]fluorometaraminol (4-[18F]FMR), a tracer for cardiac sympathetic innervation, was synthesized by electrophilic aromatic substitution. A trimethylstannyl precursor, protected with tert-butoxycarbonyl protecting groups, was radiofluorinated with high specific radioactivity [18F]F2. Specific radioactivity of 4-[18F]FMR, in average 11.8 +/-3.3 GBq/micromol, was improved 40 800-fold in comparison to the previous electrophilic fluorinations. The biodistribution of 4-[18F]FMR in rat was in accordance with the known distribution of sympathetic innervation. 4-[18F]FMR showed no metabolic degradation in left ventricle of rat heart, where the uptake was high, rapid and specific. PMID- 14741576 TI - Synthesis and evaluation of iodinated TZTP-derivatives as potential radioligands for imaging of muscarinic M2 receptors with SPET. AB - A series of iodinated thiadiazolyltetrahydro-1-methyl-pyridine (TZTP) compounds was synthesized and evaluated in vitro and in vivo as potential radioligands for imaging of the muscarinic M2 receptor subtype with SPET. One of these compounds, 5-(E)-iodopentenylthio-TZTP, has high in vitro affinity (Ki = 4.9 nM) and moderate selectivity for the muscarinic M2 receptor subtype. Although the uptake pattern in the biodistribution studies in rats is consistent with muscarinic M2 receptor disribution, specific in vivo binding to these receptors could not be demonstrated. The usefulness of this tracer in human SPET imaging may therefore be limited. PMID- 14741577 TI - Radiosynthesis of [18F] N-(3-Fluoropropyl)-2-beta-Carbomethoxy-3-beta-(4 Bromophenyl) Nortropane and the regional brain uptake in non human primate using PET. AB - A synthetic procedure for the preparation of [18F]FPCBT, an imaging agent for the dopamine transporter (DAT), has been developed. The radiosynthesis was carried out in a two step procedure. Even though the yield was low, we were able to prepare 20 to 30 mCi of the product, which was enough for two or three studies. The radiochemical purity was greater than 96%. The in vivo properties of this radiotracer were evaluated using baboon and it showed highest uptake in the striatum. The studies also revealed that the maximum uptake was reached within 7 to 10 minutes post injection. Plasma metabolite analysis indicated that there is only one metabolite and it is less lipophilic than the parent compound. [18F]FPCBT displayed good brain uptake and its high target to non target ratio indicate that it is a potential candidate for DAT imaging. PMID- 14741578 TI - Rapid and reproducible radiosynthesis of [18F] FHBG. AB - 9-(4-[18F] Fluoro-3-hydroxymethylbutyl) guanine ([18F] FHBG), an imaging agent for gene therapy using PET, was prepared in a one-pot, two-step synthesis. Microwave (MW) mediated nucleophilic fluorination of N2, monomethoxytrityl-9-[4 (tosyl)-3-monomethoxytrityl-methylbutyl] guanine using no-carrier-added [18F] fluoride, followed by deprotection with hydrochloric acid and HPLC purification, gave [18F] FHBG. The radiochemical yield (decay corrected) was 12+/-5% (n = 35), the synthesis time was 55-60 min, and the radiochemical purity was >99%. The compound was used for lung imaging and was injected into Sprague-Dawley rats previously infected with the herpes simplex virus type 1 thymidine kinase (HSV1 tk) reporter gene. MicroPET imaging showed accumulation confined to the lungs. PMID- 14741579 TI - Novel 3+1 mixed-ligand Technetium-99m complexes carrying dipeptides as monodentate ligands. AB - Novel mixed ligand oxotechnetium complexes of the type [99mTcO(SSS)(SR)], in which the SR monodentate ligand is derived from dipeptides gly-gly, phe-gly and ala-gly, have been synthesized. These complexes, which have a molecular weight above 300, a lipophilic moiety, [TcO(SSS)]+, and an ionizable group separated from the lipophilic moiety by a spacer, have been obtained in 70-95% radiochemical yield. These compounds were prepared using 99mTc-tartrate as the precursor and Sn2+ as the reducing agent. The identity of the [99mTcO(SSS)(SR)] complexes has been established by HPLC comparison with the analogous oxorhenium complexes. The nature of the monodentate co-ligand strongly affects the stability of the 99mTc-complexes and their biodistribution. Complex 3b is the most stable in vitro presenting the highest blood clearance, a high liver uptake and a selective hepatobiliary excretion (54.5% ID at 15 min post-injection, and 69.3% ID at 60 min post injection). The results obtained show that 3b have reasonable stability and in vivo properties that may be useful for peptide labeling. PMID- 14741580 TI - The gross structure of the respiratory complex I: a Lego System. AB - The proton-pumping NADH:ubiquinone oxidoreductase, also called complex I, is the entry point for electrons into the respiratory chains of many bacteria and mitochondria of most eucaryotes. It couples electron transfer with the translocation of protons across the membrane, thus providing the proton motive force essential for energy-consuming processes. Electron microscopy revealed the 'L'-shaped structure of the bacterial and mitochondrial complex with two arms arranged perpendicular to each other. Recently, we showed that the Escherichia coli complex I takes on another stable conformation with the two arms arranged side by side resulting in a horseshoe-shaped structure. This model reflects the evolution of complex I from pre-existing modules for electron transfer and proton translocation. PMID- 14741581 TI - Disruption of a specific molecular interaction with a bound lipid affects the thermal stability of the purple bacterial reaction centre. AB - Relatively little is known about the functions of specific molecular interactions between membrane proteins and membrane lipids. The structural and functional consequences of disrupting a previously identified interaction between a molecule of the diacidic lipid cardiolipin and the purple bacterial reaction centre were examined. Mutagenesis of a highly conserved arginine (M267) that is responsible for binding the head-group of the cardiolipin (to leucine) did not affect the rate of photosynthetic growth, the functional properties of the reaction centre, or the X-ray crystal structure of the complex (determined to a resolution of 2.8 A). However, the thermal stability of the protein was compromised by this mutation, part of the reaction centre population showing an approximately 5 degrees C decrease in melting temperature in response to the arginine to leucine mutation. The crystallised mutant reaction centre also no longer bound detectable amounts of cardiolipin at this site. Taken together, these observations suggest that this particular protein-lipid interaction contributes to the thermal stability of the complex, at least when in detergent micelles. These findings are discussed in the light of proposals concerning the unfolding processes that occur when membrane proteins are heated, and we propose that one function of the cardiolipin is to stabilise the interaction between adjacent membrane-spanning alpha-helices in a region where there are no direct protein-protein interactions. PMID- 14741582 TI - Heme protein films with polyamidoamine dendrimer: direct electrochemistry and electrocatalysis. AB - Biocompatible nanosized polyamidoamine (PAMAM) dendrimer films provided a suitable microenvironment for heme proteins to transfer electron directly with underlying pyrolytic graphite (PG) electrodes. Hemoglobin (Hb), myoglobin (Mb), horseradish peroxidase (HRP), and catalase (Cat) incorporated in PAMAM films exhibited a pair of well-defined, quasi-reversible cyclic voltammetric peaks, respectively, characteristic of the protein heme Fe(III)/Fe(II) redox couples. While Hb-, Mb-, and HRP-PAMAM films showed the cyclic voltammetry (CV) peaks at about -0.34 V vs. saturated calomel electrode (SCE) in pH 7.0 buffers, Cat-PAMAM films displayed the peak pair at a more negative potential of -0.47 V. The protein-PAMAM films demonstrated a surface-confined or thin-layer voltammetric behavior. The electrochemical parameters such as apparent heterogeneous electron transfer rate constants (k(s)) and formal potentials (E (degrees ')) were estimated by square wave voltammetry with nonlinear regression analysis. UV-vis and IR spectroscopy showed that the proteins retained their near-native secondary structures in PAMAM films. Oxygen, hydrogen peroxide, and nitrite were catalytically reduced at the protein-PAMAM film electrodes, showing the potential applicability of the films as the new type of biosensors or bioreactors based on direct electrochemistry of the proteins. PMID- 14741583 TI - Energy transfer in monomeric phycoerythrocyanin. AB - Phycoerythrocyanin (PEC) is part of the phycobilisome of cyanobacteria. Its monomer carries one phycoviolobilin and two phycocyanobilins (PCB) as chromophores. For an understanding of the complicated energy transfer in phycobilisomes, a detailed knowledge of the processes in the constituting building proteins is indispensable. We report the experimental data necessary for the description of Forster energy transfer in monomeric PEC, including fluorescence lifetimes and quantum yields of the two subunits. The bulk experiments are complemented by studies of single PEC molecules. Forster energy calculations and Monte Carlo simulations based on the bulk data are presented. They reveal that earlier experimental findings of energy transfer heterogeneities in single PEC molecules originate in spectral shifts between the contributing chromophores. PMID- 14741584 TI - The main external alternative NAD(P)H dehydrogenase of Neurospora crassa mitochondria. AB - A DNA sequence homologous to non-proton-pumping NADH dehydrogenase genes was found in the genome of Neurospora crassa encoding a polypeptide of 577 amino acid residues, molecular mass of 64,656 Da, with a putative transmembrane domain. Analysis of fungal mitochondria fractionated with digitonin indicates that the protein is located at the outer face of the inner membrane of the organelle (external enzyme). The corresponding gene was inactivated by the generation of repeat-induced point mutations. Mitochondria from the resulting null-mutant nde2 are highly deficient in the oxidation of cytosolic NADH and NADPH. A triple mutant nde1/nde2/ndi1, lacking mitochondrial alternative NAD(P)H dehydrogenases, was obtained, indicating that these proteins are not essential in N. crassa. However, crosses between the nde2 mutant strain and complex I-deficient mutants yielded no viable double mutants. Transcription of the nde-2 gene, as well as of ndi-1 (internal enzyme), is repressed in the late exponential phase of fungal growth. PMID- 14741585 TI - Quantification of photosystem I and II in different parts of the thylakoid membrane from spinach. AB - Electron paramagnetic resonance (EPR) was used to quantify Photosystem I (PSI) and PSII in vesicles originating from a series of well-defined but different domains of the thylakoid membrane in spinach prepared by non-detergent techniques. Thylakoids from spinach were fragmented by sonication and separated by aqueous polymer two-phase partitioning into vesicles originating from grana and stroma lamellae. The grana vesicles were further sonicated and separated into two vesicle preparations originating from the grana margins and the appressed domains of grana (the grana core), respectively. PSI and PSII were determined in the same samples from the maximal size of the EPR signal from P700(+) and Y(D)( . ), respectively. The following PSI/PSII ratios were found: thylakoids, 1.13; grana vesicles, 0.43; grana core, 0.25; grana margins, 1.28; stroma lamellae 3.10. In a sub-fraction of the stroma lamellae, denoted Y-100, PSI was highly enriched and the PSI/PSII ratio was 13. The antenna size of the respective photosystems was calculated from the experimental data and the assumption that a PSII center in the stroma lamellae (PSIIbeta) has an antenna size of 100 Chl. This gave the following results: PSI in grana margins (PSIalpha) 300, PSI (PSIbeta) in stroma lamellae 214, PSII in grana core (PSIIalpha) 280. The results suggest that PSI in grana margins have two additional light-harvesting complex II (LHCII) trimers per reaction center compared to PSI in stroma lamellae, and that PSII in grana has four LHCII trimers per monomer compared to PSII in stroma lamellae. Calculation of the total chlorophyll associated with PSI and PSII, respectively, suggests that more chlorophyll (about 10%) is associated with PSI than with PSII. PMID- 14741586 TI - Formation of radicals from singlet oxygen produced during photoinhibition of isolated light-harvesting proteins of photosystem II. AB - Electron spin resonance spectroscopy and liquid chromatography have been used to detect radical formation and fragmentation of polypeptides during photoinhibition of purified major antenna proteins, free of protease contaminants. In the absence of oxygen and light, no radicals were observed and there was no damage to the proteins. Similarly illumination of the apoproteins did not induce any polypeptide fragmentation, suggesting that chlorophyll, light and atmospheric oxygen are all participating in antenna degradation. The use of TEMP and DMPO as spin traps showed that protein damage initiates with generation of (1)O(2), presumably from a triplet chlorophyll, acting as a Type II photosensitizer which attacks directly the amino acids causing a complete degradation of protein into small fragments, without the contribution of proteases. Through the use of scavengers, it was shown that superoxide and H(2)O(2) were not involved initially in the reaction mechanism. A higher production of radicals was observed in trimers than in monomeric antenna, while radical production is strongly reduced when antennae were organized in the photosystem II (PSII) complex. Thus, monomerization of antennae as well as their incorporation into the PSII complex seem to represent physiologically protected forms. A comparison is made of the photoinhibition mechanisms of different photosynthetic systems. PMID- 14741587 TI - The proliferation and phenotypic expression of human osteoblasts on tantalum metal. AB - Tantalum (Ta) is increasingly used in orthopaedics, although there is a paucity of information on the interaction of human osteoblasts with this material. We investigated the ability of Ta to support the growth and function of normal human osteoblast-like cells (NHBC). Cell responses to polished and textured Ta discs were compared with responses to other common orthopaedic metals, titanium and cobalt-chromium alloy, and tissue culture plastic. No consistent differences, that could be attributed to the different metal substrates or to the surface texture, were found in several measured parameters. Attachment of NHBC to each substrate was similar, as was cell morphology, as determined by confocal microscopy. Cell proliferation was slightly faster on plastic than on Ta at 3 days, but by 7 days neither the absolute cell numbers, nor the number of cell divisions, was different between Ta and the other substrates. No consistent, substrate-dependent differences were seen in the expression of a number of mRNA species corresponding to the pro-osteoclastic or the osteogenic activity of osteoblasts. No substrate-dependent differences were seen in the extent of in vitro mineralisation by NHBC. These results indicate that Ta is a good substrate for the attachment, growth and differentiated function of human osteoblasts. PMID- 14741588 TI - Chronic response of adult rat brain tissue to implants anchored to the skull. AB - Using quantitative immunohistological methods, we examined the brain tissue response to hollow fiber membranes (HFMs) that were either implanted intraparenchymally, as in a cell encapsulation application, or were attached to the skull as in a biosensor application (transcranially). We found that the reaction surrounding transcranially implanted HFMs was significantly greater than that observed with intraparenchymally implanted materials including increases in immunoreactivity against GFAP, vimentin, ED-1 labeled macrophages and microglia, and several extracellular matrix proteins including collagen, fibronectin, and laminin. In general, these markers were elevated along the entire length of transcranially implanted HFMs extending into the adjacent parenchyma up to 0.5 mm from the implant interface. Intraparenchymal implants did not appear to have significant involvement of a fibroblastic component as suggested by a decreased expression of vimentin, fibronectin and collagen-type I at the implant tissue interface. The increase in tissue reactivity observed with transcranially implanted HFMs may be influenced by several mechanisms including chronic contact with the meninges and possibly motion of the device within brain tissue. Broadly speaking, our results suggest that any biomaterial, biosensor or device that is anchored to the skull and in chronic contact with meningeal tissue will have a higher level of tissue reactivity than the same material completely implanted within brain tissue. PMID- 14741589 TI - Early detachment of titanium particles from various different surfaces of endosseous dental implants. AB - Titanium (Ti) endosseous dental screws with different surfaces (smooth titanium- STi, titanium plasma-sprayed-TPS, alumina oxide sandblasted and acid-etched--Al SLA, zirconium oxide sandblasted and acid etched--Zr-SLA) were implanted in femura and tibiae of sheep to investigate the biological evolution of the peri implant tissues and detachment of Ti debris from the implant surfaces in early healing. Implants were not loaded. Sections of the screws and the peri-implant tissues obtained by sawing and grinding were analysed by light microscopy immediately after implantation (time 0) and after 14 days. All samples showed new bone trabeculae and vascularised medullary spaces in those areas where gaps between the implants and host bone were visible. In contrast, no osteogenesis was induced in the areas where the implants were initially positioned in close contact with the host bone. Chips of the pre-existing bone inducing new peri implant neo-osteogenesis were surrounded by new bone trabeculae. The threads of some screws appeared to be deformed where the host bone showed fractures. Ti granules of 3-60 microm were detectable only in the peri-implant tissues of TPS implants both immediately after surgery and after 14 days, thus suggesting that this phenomenon may be related to the friction of the TPS coating during surgical insertion. PMID- 14741590 TI - Protein and bacterial fouling characteristics of peptide and antibody decorated surfaces of PEG-poly(acrylic acid) co-polymers. AB - The potential for base poly(ethylene glycol) graft poly(acrylic acid) PEG-g-PA copolymers and surface-modified PEG-g-PA materials to inhibit random protein fouling and bacterial adhesion are investigated. PEG-g-PA co-polymers were synthesized that inhibited non-specific protein and cellular adhesion. PEG-g-PA co-polymers were then covalently modified with either cell adhesion peptides (YRGDS, YEILDV) or fragments of antibodies to monocyte/macrophage integrin receptors (Anti-VLA4, Anti-beta1, Anti-beta2, and Anti-CD64) known to enhance macrophage adhesion and, perhaps, modulate their activation. Materials produced in this work were characterized using: hydrophobicity by contact angle; angle resolved X-ray Photoelectron Spectroscopy to confirm the presence of PEG in the bulk material and the surface; degree of hydration; differential scanning calorimetry; and thermal gravimetric analysis. To evaluate the non-fouling efficacy of the various modified surfaces, three proteins, human serum albumin, human fibronectin (Fraction I) and human immunoglobulin were 125I labeled. Samples of base PEG-g-PA and PEG-g-PA, modified with various peptides, were exposed to solutions containing either 2 or 200 microg/ml of one of the labeled proteins at 37 degrees C for 24 h. PEG-g-PA substrata modified with directly bound peptides exhibited protein adsorption that varied depending upon the surface bounded peptide. PEG-g-PA modified with peptides linked by linear PEG tethers reduced protein adsorption at 24 h by approximately 45% in comparison to PEG-g-PA. Peptides linked by way of StarPEO and StarlikePEO tethers further decreased protein adsorption in comparison to PEG-g-PA. The ability of peptide:PEOtethers to inhibit protein adsorption appeared to be a function of type and surface coverage of the PEO tether and not influenced by the amount or molecular structure the tethered peptide. Peptides directly coupled to the PEG-g PA increased the amount of protein fouling relative to controls and there appeared to be some dependency of the amount of protein adsorption on which peptide was tethered. Two 14C-labeled pathogens, Staphylococcus epidermidis and Pseudomonas aeruginosa, were used to quantify the degree of bacterial adhesion using two types of laminar flow cell chambers; one that provided invasive sampling of the target substrata and one that provided non-invasive microscopic surveillance of adhering bacterial cells. Attachment of both species to PEG-g-PA and peptide-modified PEG-g-PA was reduced compared to the basic poly(acrylic acid). Presence of peptides on the surface, whether directly bound or bound by the PEO tether did not influence adhesion of P. aeruginosa relative to controls. S. epidermidis adhesion rates increased slightly for those materials where peptides were directly bound to the surface but were reduced relative to base PEG g-PA when peptides were bound by PEO tethers. All PEG-g-PA surfaces modified with fragments of monoclonal antibodies dramatically enhanced bacterial initial adhesion rates and maximum extent of attachment. PMID- 14741591 TI - Preparation and physicochemical properties of compression-molded keratin films. AB - The S-sulfo keratin was extracted from wool and was then spray-dried to give S sulfo keratin powder. Differential scanning calorimetry analysis showed that the glass transition temperature of S-sulfo keratins became lowered with the increase of moisture content, while perfectly dried S-sulfo keratin powder did not give thermal transition in the temperature range 30-130 degrees C. The compression molding of the S-sulfo keratin powder supplemented with one-tenth weight of water afforded a plastic-like transparent proteinous film above the glass transition temperature. The film obtained from the powder without water addition or compression molded below glass transition temperature partly remained powdery. The film compression molded at 120 degrees C gave the maximum ultimate strength and Young's modulus, 27.8 +/- 2.9 and 1218 +/- 80 MPa, respectively. Obtained film was insoluble and slightly swelled in water, but, in the presence of reducing agent, the film significantly swelled at pH 7.0 and even dissolved at pH 9.0, suggesting the relevance of abundant disulfide linkage. The film supported the mammalian cell adhesion and proliferation, demonstrating the biocompatibility of S-sulfo keratin films. PMID- 14741592 TI - Silicone elastomers for reduced protein adsorption. AB - Monofunctional poly(ethylene oxide) polymers of molecular weight (MW) 350, 750, and 2000, respectively, were modified with Si(OEt)3 groups. These polymers underwent classic condensation cure with hydroxy-terminated silicone polymers and Si(OEt)4 to give composites with poly(ethylene oxide) (PEO) rich surfaces under aqueous conditions, as shown by contact angle and XPS data. The hydrophobicity of the surfaces was considerably higher in air. The greatest PEO concentration was observed with relatively short chain polymers of MW 350. Silicone polymers bearing short chain PEO chains were also observed to be the most protein rejecting from either buffer (fibrinogen) (90%) or plasma (85%). The silicone/TES MPEO formulation offers the advantage of a one step/one shot polymerization process that gives materials with a high protein rejection ability than can be cast as films, or molded into complex shapes. Covalently linked PEO films of a variety of chain lengths and total surface coverage can be readily accommodated. PMID- 14741593 TI - Soluble phosphate glasses: in vitro studies using human cells of hard and soft tissue origin. AB - This report describes the short-term response of two typical cellular components of a hard/soft tissue interface such as the periodontal ligament/mandible and patellar tendon/tibia. Tissue engineering of such interfaces requires a contiguous scaffold system with at least two cell types associated with it. Human oral osteoblasts, oral fibroblasts and hand flexor tendon fibroblasts were seeded on phosphate-based glasses of different dissolution rates. Quantitative and morphological assessment of cell adhesion and proliferation for all cell types was assessed, after first elucidating an experimental composition range using MG63 cells. In addition, immunolabelling of bone-specific non-collagenous proteins bone sialoprotein, osteonectin and osteopontin was performed to determine osteoblast phenotype. Fibroblast phenotype was established by immunolabelling for prolyl-4-hydroxylase, an enzyme vital for collagen biosynthesis. Results indicated that both cell types maintained their respective phenotypes over time in culture on glass discs of generic composition (CaO)x (Na2O)(0.5-x)-(P2O5)0.5, remained attached and proliferated dependent on glass composition and cell type. Glasses containing at least 46 mol% CaO, produced no adverse cell reaction suggesting that these compositions that support both osteoblasts and fibroblasts would be ideal as a scaffold material for engineering the hard/soft tissue interface. PMID- 14741594 TI - Immobilization of erythropoietin to culture erythropoietin-dependent human leukemia cell line. AB - To investigate the effect of immobilized cytokine, erythropoietin (Epo) was immobilized on a culture plate and the Epo-dependent human leukemia cell line UT 7/Epo then was cultured upon the plate. A photo-reactive gelatin was mixed with Epo and the mixture was cast on a plate. The plate was then irradiated with ultraviolet light in the presence or absence of a photo-mask. After washing with water, a micropatterned or unpatterned surface was formed. A leukemia cell line dependent on Epo, UT-7/Epo, was cultured on the sample plate. On the micropatterned surface, apoptosis of cells was induced on the surface without Epo, but was not observed on the Epo-immobilized surface. This result demonstrated that Epo stimulated the cells even after immobilization. Although the activity of immobilized Epo was low, the activity was slightly higher than that achieved by soluble Epo at higher concentration. In addition, the immobilized Epo could be repeatedly used for culture of UT-7/Epo cell. The present study provided a convenient immobilization method and indicated that immobilization of cytokines will be useful for creating an artificial cell culture device. PMID- 14741595 TI - Differentiation, growth and activity of rat bone marrow stromal cells on resorbable poly(L/DL-lactide) membranes. AB - Nonporous and porous membranes produced from poly(L/DL-lactide) 80/20% were characterized using profilometry, contact-angle measurements, infra-red spectroscopy, X-ray photoemission spectroscopy and scanning electron microscopy, and used to culture bone marrow stromal cells isolated from the rat femora. The cells were cultured for 5, 10, 15 and 20 days. Cell growth and activity was estimated from the amounts of DNA, alkaline phosphatase activity and total protein amount present in the cell lysate and cell differentiation was assessed histochemically. Cell morphology was estimated from scanning electron microscopy. The cells fully expressed osteoblastic phenotype, revealed spindle-shaped, ellipsoidal morphology, developed podia, produced an abundant fibrillar extracellular matrix and mineral noduli. The number of cells on the membranes increased with time of culturing and was higher for the porous membranes than the nonporous membranes. Osteoblastic differentiation was most significant between 5 and 10 days of culture. The total amounts of DNA, alkaline phosphatase and proteins increased with time of culturing. The surface characteristics of the porous membranes were superior to the nonporous membranes. PMID- 14741596 TI - Preparation and characteristics of hybrid scaffolds composed of beta-chitin and collagen. AB - Hybrid scaffolds composed of beta-chitin and collagen were prepared by combining salt-leaching and freeze-drying methods. The chitin scaffold used as a framework was easily formed into desired shapes with a uniformly distributed and interconnected pore structure with average pore size of 260-330 microm. The mechanical strength and the rate of biodegradation increased with the porosity, which could be modulated by the salt concentration. In addition, atelocollagen solution was introduced into the macropores of the chitin scaffold to improve cell attachment. Web-like collagen fibers fabricated between pores of chitin were produced by a 0.1 wt% collagen solution, whereas a 0.5 wt% collagen solution only coated the surface of the chitin scaffold. After 3 days of culture, fibroblasts cultured in collagen-coated scaffolds were attached at the place where the collagen was fabricated, whereas cells did not attach and aggregate on the scaffold of chitin alone. After 14 days, the fibroblasts showed a good affinity to and proliferation on all collagen-coated chitins. PMID- 14741597 TI - Effect of PEG-PLLA diblock copolymer on macroporous PLLA scaffolds by thermally induced phase separation. AB - A regular and highly interconnected macroporous poly(L-lactic acid) (PLLA) scaffold was fabricated from a PLLA-dioxane-water ternary system with added polyethylene glycol (PEG)-PLLA diblock using thermally induced phase separation (TIPS). The morphology of the scaffold was investigated in detail by controlling the following TIPS parameters: quenching temperature, aging time, polymer concentration, molecular structure, and diblock concentration. The phase diagram was assessed visually on the basis of the turbidity. The cloud-point curve shifted to higher temperatures with increasing PEG content in the additives (PEG PLLA diblocks), due to a stronger interaction between PEG and water in solution. The addition of diblock series (0.5 wt% in solution) stabilized interconnections of pores at a later stage without segregation or sedimentation. The pore size of the scaffold could be easily controlled in the range 50-300 microm. A macroporous PLLA scaffold was used to study an MC3T3-E1 cell (an osteoblast-like cell) culture. The cells successfully proliferated in the PLLA scaffold in the presence of added PEG-PLLA diblock for 4 weeks. PMID- 14741598 TI - CaO--P2O5--Na2O-based sintering additives for hydroxyapatite (HAp) ceramics. AB - We have assessed the effect of CaO--P2O5--Na2O-based sintering additives on mechanical and biological properties of hydroxyapatite (HAp) ceramics. Five different compositions of sintering additives were selected and prepared by mixing of CaO, P2O5, and Na2CO3 powders. 2.5 wt% of each additive was combined with commercial HAp powder, separately, followed by ball milling, and sintering at 1250 degrees C and 1300 degrees C in a muffle furnace. Green and sintered densities of the compacts were analyzed for the influence of additives on densification of HAp. Phase analyses were carried out using an X-ray diffractometer. Vickers microhardness testing was used to evaluate hardness of sintered compacts of different compositions. A maximum microhardness of 4.6 (+/- 0.28) GPa was attained for a composition with 2.5 wt% addition of CaO:P2O5:Na2O in the ratio of 3:3:4. Results from mechanical property evaluation showed that some of these sintering additives improved failure strength of HAp under compressive loading. Maximum compressive strength was observed for samples with 2.5 wt% addition of CaO. Average failure strength for this set of samples was calculated to be 220 (+/- 50) MPa. Cytotoxicity, and cell attachment studies were carried out using a modified human osteoblast cell line called OPC-1. In vitro results showed that these compositions were non-toxic. Some sintering aids enhanced cell attachment and proliferation, which was revealed from SEM examination of the scaffolds seeded with OPC-1 cells. PMID- 14741599 TI - Protein adsorption, attachment, growth and activity of primary rat osteoblasts on polylactide membranes with defined surface characteristics. AB - The adsorption of proteins and growth and activity of primary rat osteoblasts cultured for 1, 2 and 3 weeks on nonporous and porous resorbable poly(L/DL lactide) 80/20% membranes with defined surface characteristics were investigated. The growth and activity of cells were estimated from the measurements of DNA, alkaline phosphatase activity and the total amount of protein in the cell lysate. The cell morphology was assessed from scanning electron microscopy and rhodamine staining. The protein adsorption to the membrane surface was assessed from the amide I peak at 1640-1660 cm(-1) and the amide II peak at 1540-1560 cm(-1) in the attenuated total reflection infrared spectra. The relative amount of proteins adsorbed on the nonporous and porous membranes was comparable. The cells growing on the nonporous and porous membranes maintained the phenotype and revealed morphology typical for osteoblasts. The mineralized noduli were larger in size on the porous membranes. The number of cells, the amount of DNA, the alkaline phosphatase activity, and the total amount of protein increased with time of the experiment and were higher for the porous membranes than for the nonporous ones. PMID- 14741600 TI - Biaxial strength of multilaminated extracellular matrix scaffolds. AB - Xenogeneic extracellular matrix (ECM) can be harvested and configured to function as a bioscaffold for tissue and organ reconstruction. The mechanical properties of the ECM vary depending upon the tissue from which it is harvested. Likewise, the manufacturing steps required to develop ECMs into medical grade devices will affect the surface morphology and the mechanical properties of the bioscaffold; important properties for constructive tissue remodeling. The present study compared the ball-burst strength of five different ECM scaffolds before and after treatment with peracetic acid (PAA): porcine small intestinal submucosa (SIS), porcine urinary bladder submucosa (UBS), porcine urinary bladder matrix (UBM), a composite of UBS + UBM, and canine stomach submucosa (SS). This study also compared the mechanical properties of 2- and 4-layer ECM scaffolds. Results showed 2-layer SS devices had the highest ball-burst value of all 2-layer ECM devices. Moreover, all 4-layer ECM devices had similar ball-burst strength except for 4-layer UBM devices which was the weakest. PAA-treatment decreased the ball burst strength of SS and increased the ball-burst strength of UBS 2-layer devices. This study showed the material properties of the ECM scaffolds could be engineered to mimic those of native soft tissues (i.e. vascular, musculotendinous, etc) by varying the number of layers and modifying the disinfection/sterilization treatments used for manufacturing. PMID- 14741601 TI - Characteristics of tissue-engineered cartilage from human auricular chondrocytes. AB - This study was done to define the mechanical and histological properties of tissue-engineered cartilage (TEC) derived from human chondrocytes and to compare these findings with those of native cartilage. Chondrocytes were obtained from 10 human auricular cartilages and seeded onto a biodegradable template of polyglycolic acid and poly L-lactic acid. Each template was shaped into a 1 cm x 2 cm rectangle. The templates were implanted in athymic mice for 8 weeks. Eight human auricular cartilages were used for comparison. Mechanical analysis with a tensile testing device provided values of ultimate tensile strength (UTS), stiffness, and resilience. Statistical analysis was performed with the Student's t-test. Histological assessment was done with hematoxylin-eosin staining along with other special stains. The TEC had UTS of 2.07 MPa, stiffness of 3.7 MPa, and resilience of 0.37 J/m3. The control specimens had UTS of 2.18 MPa, stiffness of 5.11 MPa, and resilience of 0.42 J/m3. No statistical difference was found between the experimental and control groups for each of the three parameters. Histological analysis showed mature cartilage with characteristic collagen, glycosaminoglycans, and elastin in the TEC. The neo-cartilage showed slightly smaller size and more irregular distribution of chondrocytes and unique fibrous capsule formation with peripheral infiltration of fibrous tissue. This study showed that the mechanical qualities of TEC from human chondrocytes are similar to those of native auricular cartilage. It suggests that the engineered cartilage from human chondrocytes may have sufficient strength and durability for clinical uses. The histological findings revealed some differences with neo-cartilage. PMID- 14741602 TI - Mineralization behaviour of collagen type I immobilized on different substrates. AB - Collagen type I as a robust fibre protein and main component of the extracellular matrix of most tissues is increasingly utilized for surface engineering of biomaterials using different immobilization methods. In the present work we studied the mineralization behaviour of fibrillar collagen type I in simulated body fluid as a measure for conformational changes caused by adsorptive immobilization or immobilization by partial incorporation into the anodic oxide layer on c.p.-titanium using microscopic and vibration spectroscopic methods. Adsorptive immobilization on highly oriented pyrolytic graphite (HOPG) and c.p. titanium without collagen were used as references. In the initial phase (1-24 h) the kinetics of formation and the morphology of calcium phosphate phases (CPP) are strongly influenced both by the substrate and the immobilization method. Compared to HOPG both types of immobilization on titanium increasingly inhibit the formation of CPP. For longer times (30 d) these initial differences disappear mineralization product on titanium, irrespective of the presence of collagen, is a mixture of amorphous calcium phosphate and octacalcium phosphate. Contrary to this the mineralization of HOPG substrates results in hydroxy apatite. This is discussed with respect to the conditions during the immobilization as well as the resulting interactions between substrate and immobilized collagen. It is shown that the mineralization process exhibits a high sensitivity with respect to conformational changes caused by these interactions. Possible cell biological relevance of these conformational changes is discussed. PMID- 14741603 TI - Injectable self-curing bioactive acrylic-glass composites charged with specific anti-inflammatory/analgesic agent. AB - Injectable bioactive acrylic formulations based on poly(methyl methacrylate) (PMMA) and different amounts of bioactive glasses in the system SiO2-CaO-Na2O P2O5 have been prepared in the presence of the anti-inflammatory analgesic drug fosfosal, the sodium salt of 2-phosphonoxibenzoic acid, to be used in minimally invasive surgery. The injectability of the formulations evaluated according to the established protocol was around 80%. The experimental formulations provided maximum temperatures in the range 50-60 degrees C, which were lower than those of commercial acrylic bone cements currently used in percutaneous vertebroplasty (PVP). Residual monomer content of any formulation was inferior to 5%. Compressive yield strength of dry specimens was in the range 80-95 MPa, but it decreased after immersion in SBF to values in the range 30-50 MPa, due to the dissolution of the bioactive glasses and the drug in the medium. The release of fosfosal was evaluated in vitro (pH = 7.0). The release profile against time obtained from a PMMA cement was quasi-linear and the 80% of the initial amount of drug was released in 175 h. However, for bioactive cements, the 80-100% of the fosfosal charged was released in approximately 48 h, due to the dissolution of the glasses in the medium. Values of weight loss of the cements determined gravimetrically ranged between 16% and 26% depending on the initial amount of fosfosal, i.e. 20 or 30 wt%, respectively. The weight loss and the water uptake were simultaneous processes, and values of hydration degree were around 10-14%. The formation of an apatite-like layer was detected on the surface of the cements at different periods of time depending on the composition of the bioactive glasses. The cements containing the glasses with P2O5 produced the growth of the apatite layer in shorter periods of time. The presence of fosfosal accelerated the precipitation of this layer independently on the glasses. The in vivo biocompatibility studied by intramuscular implantation in rats showed the absence of an anti-inflammatory response and a fibrous layer around the implant for the cement prepared with PMMA/fosfosal which is attributed to the therapeutic action of fosfosal acting in situ. The response to cements prepared with bioactive glasses and fosfosal showed a mild inflammatory reaction with the formation of the typical fibrous capsule around the implanted material. PMID- 14741604 TI - Hydrogen-bonded polymer gel and its application as a temperature-sensitive drug delivery system. AB - The mixture of poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) triblock copolymer (F-68) and poly vinyl alcohol (PVA) forms a polymer complex gel by intra/intermolecular interaction via hydrogen bonding in water, which is verified by differential scanning calorimetry. With 30 wt% F-68 aqueous solution and 15 wt% PVA aqueous solution, F-68/PVA complex gel was prepared and its swelling transition was observed at approximately 37 degrees C. Based on the temperature-sensitivity of hydrogen bondings in F-68/PVA complex gel, temperature sensitive drug delivery system has been designed and characterized. For the stability in the aqueous media, F-68/PVA complex gel was prepared with a form of polymeric bead, followed by the encapsulation with poly(lactide-co-glycolide) membrane. With changing the ratio of F-68/PVA, the swelling transition of polymer complex gel was manipulated and pulsatile release of acetoaminophen, used as a model drug, was demonstrated in response to pulsatile change of temperature between 35 degrees C and 40 degrees C. PMID- 14741605 TI - Bioadhesive, non-drug-loaded nanoparticles as modulators of candidal adherence to buccal epithelial cells: a potentially novel prophylaxis for candidosis. AB - Adherence of microorganisms, such as Candida albicans, represents the initial step in the establishment of infection and, accordingly, modification of this step represents a method by which the incidence of infection may be reduced. Therefore, this study uniquely examined the effects of polymeric nanoparticles on the adherence of blastospores of C. albicans to human buccal epithelial cells (BEC) in vitro. Poly(propylcyanoacrylate) nanoparticles were produced by emulsion polymerisation using a range of anionic, cationic and non-ionic surfactants, their particle size and zeta potential characterised and incubated with stationary phase blastospores of C. albicans for a defined period. Following this, the surface properties and size of blastospores with adsorbed nanoparticles were characterised. phosphate buffered saline-treated and nanoparticle-treated blastospores were incubated with human BEC for 2 h, following which the number of adherent blastospores was enumerated by light microscopy. The size and zeta potential of the nanoparticles were dependent on the surfactant employed in the manufacture process. Following nanoparticle adsorption, alteration of the zeta potential and an increase in the diameter of blastospores were observed. However, as this increase in diameter was indirectly related to the size of the nanoparticles, this may indicate a preference for the adsorption of smaller particles. In addition, following nanoparticle adsorption, the cell surface hydrophobicity (CSH) of C. albicans blastospores was increased and, importantly, the subsequent adherence to BEC in vitro was reduced. Most notably, the adherence of blastospores that had been treated with nanoparticles (stabilised with docusate sodium) was circa 73% lower than that of untreated blastospores. A moderate correlation between increased CSH and reduced adherence and a low correlation between blastospore zeta potential and adherence were observed, inferring that other mechanisms, most likely stearic hindrance, are responsible for the antiadherent properties of adsorbed nanoparticles. In light of their ability to reduce candidal adherence to BEC, it is suggested that polymeric nanoparticles may be useful in the prophylaxis of candidosis of the oral cavity. PMID- 14741606 TI - The effect of the processing and formulation parameters on the size of nanoparticles based on block copolymers of poly(ethylene glycol) and poly(N isopropylacrylamide) with and without hydrolytically sensitive groups. AB - Block copolymers of poly(ethylene glycol) (PEG) as a hydrophilic block and N isopropylacrylamide (PNIPAAm) or poly (NIPAAm-co-N-(2-hydroxypropyl) methacrylamide-dilactate) (poly(NIPAAm-co-HPMAm-dilactate)) as a thermosensitive block, are able to self-assemble in water into nanoparticles above the cloud point (CP) of the thermosensitive block. The influence of processing and the formulation parameters on the size of the nanoparticles was studied using dynamic light scattering. PNIPAAm-b-PEG 2000 polymers were not suitable for the formation of small and stable particles. Block copolymers with PEG 5000 and 10000 formed relatively small and stable particles in aqueous solutions at temperatures above the CP of the thermosensitive block. Their size decreased with increasing molecular weight of the thermosensitive block, decreasing polymer concentration and using water instead of phosphate buffered saline as solvent. Extrusion and ultrasonication were inefficient methods to size down the polymeric nanoparticles. The heating rate of the polymer solutions was a dominant factor for the size of the nanoparticles. When an aqueous polymer solution was slowly heated through the CP, rather large particles (> or = 200 nm) were formed. Regardless the polymer composition, small nanoparticles (50-70 nm) with a narrow size distribution were formed, when a small volume of an aqueous polymer solution below the CP was added to a large volume of heated water. In this way the thermosensitive block copolymers rapidly pass their CP ('heat shock' procedure), resulting in small and stable nanoparticles. PMID- 14741607 TI - Hydrogen absorption behavior of beta titanium alloy in acid fluoride solutions. AB - Hydrogen absorption behavior of a beta titanium alloy in acid fluoride solutions has been analyzed by hydrogen thermal desorption. The amount of absorbed hydrogen increased with immersion time in a 2.0% acidulated phosphate fluoride (APF) solution. In the case of an immersion time of 60 h, the amount of absorbed hydrogen exceeded 10000 mass ppm. In contrast, the amount of hydrogen absorbed in the 0.2% APF solution was several times smaller than that in the 2.0% APF solution for the same immersion time. For immersion in a 0.2% APF solution, hydrogen absorption saturated after 48 h. The surface topography and corrosion products on the surface of the specimen immersed in the 2.0% APF solution were different from those in the 0.2% APF solution. During the later stage of immersion, the amount of absorbed hydrogen markedly increased under higher applied stress, although the applied stress did not enhance hydrogen absorption during the early stage of immersion. These results of hydrogen absorption behavior are consistent with the delayed fracture characteristics of the beta titanium alloy. PMID- 14741608 TI - Quasi-static and dynamic nanoindentation studies on highly crosslinked ultra-high molecular-weight polyethylene. AB - In four separate studies, the effect of three methods used to highly crosslink ultra-high-molecular-weight polyethylene (UHMWPE) (gamma irradiation, electron beam irradiation, and peroxide treatment) on the polymer's stiffness was investigated using quasi-static and dynamic nanoindentation. From the quasi static studies, it was found that (a) the highest value of the elastic modulus, E (2240 +/- 157 MPa) was obtained from specimens prepared from direct compression molded UHMWPE crosslinked using gamma irradiation, with a dosage of 7.5 Mrad; (b) the value of E for specimens formed from direct compression molded UHMWPE was higher (1950 +/- 255 MPa) compared to results obtained from specimens formed from ram extruded UHMWPE (1583 +/- 140 MPa); and (c) the magnitude of the decrease in the value of E as a result of subjecting a highly crosslinked UHMWPE specimen to uniaxial tensile forces depends on the method used for the crosslinking. From the dynamic study, (d) it was found that the extent of the change in both the storage modulus and the loss modulus, as a result of subjecting a highly crosslinked UHMWPE specimen to uniaxial tension forces, depends on the method used for the crosslinking. PMID- 14741610 TI - Thermal dimensional behavior of dental ceramics. AB - The aim of this study was to characterize the relation of dimensional changes of dental ceramic materials as function of temperature. The thermal dimensional behavior of one glass ceramic core, four commercially available veneering porcelains and two experimental veneering porcelain materials was determined on cooling from 450 degrees C to 20 degrees C by means of a vertical differential pushrod thermal dilatometer. The thermal dimensional behavior of the ceramic materials was expressed in three different ways: as linear relation with a linear line fitting procedure; as non-linear relation by a quadratic curve fitting procedure and according to ISO 9693 by a thermal expansion coefficient. For all ceramic materials except the one consisting of aluminum oxide fillers, the thermal dimensional behavior is best described with the polynomial function: Deltal/l = C + alpha1T + alpha2T2. The description of the thermal expansion coefficient according to the ISO standard on metal is not appropriate for the prediction of a thermal incompatibility of ceramic materials. Except for Vitadur Alpha, the ceramics materials tested showed non-linear expansion; their thermal dimensional behavior can be best described according to the quadratic relation method, second by the linear relation, third as recommended by ISO. PMID- 14741609 TI - Contact profilometry and correspondence analysis to correlate surface properties and cell adhesion in vitro of uncoated and coated Ti and Ti6Al4V disks. AB - A fundamental goal in the field of implantology is the design of specific devices able to induce a controlled and rapid "osseointegration". This result has been achieved by means of surface modifications aimed at optimizing implant-to-bone contact; furthermore, bone cell adhesion on implant surface has been directly improved by the application of biomolecules that stimulate new tissue formation, thus controlling interactions between biological environment and implanted materials. Actually, methods for biochemical factor delivery at the interface between implant surface and biological tissues are under investigation; a reliable technique is represented by the inclusion of biologically active molecules into biocompatible and biodegradable materials used for coating implant surface. This paper focuses the application of three polymeric materials already acknowledged in the clinical practice, i.e. poly-L-lactic acid (PLLA), poly-DL lactic acid (PDLA), and sodium alginate hydrogel. They have been used to coat Ti (Ti2) and Ti6Al4V (Ti5) disks; their characteristics have been determined and their performances compared, with specific regard to the ability in allowing osteoblast adhesion in vitro. Moreover, profilometry data analysis permitted to identify a specific roughness parameter (peak density) which mainly controls the amount of osteoblast adhesion. PMID- 14741611 TI - Hydrolytic degradation study of biodegradable polyesteramide copolymers based on epsilon-caprolactone and 11-aminoundecanoic acid. AB - In this paper, a new kind of aliphatic biodegradable polyesteramide copolymers P(CL/AU)x/y based on epsilon-caprolactone and 11-aminoundecanoic acid were synthesized by the melt polycondensation method. Hydrolytic degradation behavior of P(CL/AU) copolymers were studied by using FTIR, 1H-NMR and DSC. Chemical compositions, macromolecular weight, thickness of the test sample, and pH of the degradation medium have great effect on degradation rate. The degradation rate decreased with increase in aminoundecanoic acid content, macromolecular weight, and thickness of the test samples, but increased with incubation temperature and pH of the degradation medium. The degradation mechanism was studied according to the mathematical model developed by professor Gopferich. PMID- 14741612 TI - In vitro bioactivity of laser ablation pseudowollastonite coating. AB - Pseudowollastonite (psW) coatings on titanium alloys substrates were prepared by laser ablation and immersed in simulated body fluid (SBF) for different periods in order to investigate the nucleation and growth of hydroxyapatite (HA)-like formation on their surface. The structure of the coatings before soaking was analysed by X-ray diffraction (XRD) and scanning electron microscopy (SEM). The interfacial reactions product was examined by thin-film XRD, SEM and transmission electron microscopy at low and high resolution level, both fitted with energy dispersive X-ray spectroscopy. Additional changes in ionic concentration, using inductively couple plasma atomic emission spectroscopy, were determined as well as pH right at the psW-coatings/SBF interface using an ion-sensitive field effect transistor. The solution composition changes, increasing the Ca(2+) and Si(4+) concentration and pH as a function of the soaking time while HPO(4)(2-) decreased. The results obtained showed that the coating surfaces were covered by HA-like, which indicated that the psW-coating possesses good bioactivity and also suggested that the mechanism of HA-like layer formation in SBF was similar to that showed in in vitro test by other silica-based materials. PMID- 14741613 TI - Surface modification of ultra thin poly (epsilon-caprolactone) films using acrylic acid and collagen. AB - Poly (epsilon-caprolactone) (PCL) has been used as a bioresorbable polymer in numerous medical devices as well as for tissue engineering applications. Its main advantage is its biocompatibility and slow degradation rate. PCL surface, however, is hydrophobic and cell-biomaterial interaction is not the best. We attempt for the first time to modify an ultra thin PCL surface with collagen. The PCL film was prepared using solvent casting and biaxial stretching technique developed in our laboratory. This biaxial stretching produced an ultra thin PCL 3 7 microm thick, ideal for membrane tissue engineering applications. The PCL film was pretreated using Argon plasma, and then UV polymerized with acrylic acid (AAc). Collagen immobilization was then carried out. The modified film surface was characterized by Fourier Transform Infrared (FT-IR) and X-ray Photoelectron Spectroscopy (XPS). Water contact angles were also measured to evaluate the hydrophilicity of the modified surface. Results showed that the hydrophilicity of the surface has improved significantly after surface modification. The water contact angle dropped from 66 degrees to 32 degrees. Atomic Force Microscopy (AFM) showed an increase in roughness of the film. A change from 46 to 60 nm in the surface morphology was also observed. The effect of cells attachment on the PCL film was studied. Human dermal fibroblasts and myoblasts attachment and proliferation were improved remarkably on the modified surface. The films showed excellent cell attachment and proliferation rate. PMID- 14741614 TI - Polyelectrolyte multilayer films with pegylated polypeptides as a new type of anti-microbial protection for biomaterials. AB - Adhesion of bacteria at the surface of implanted materials is the first step in microbial infection, leading to post-surgical complications. In order to reduce this adhesion, we show that poly(L-lysine)/poly(L-glutamic acid) (PLL/PGA) multilayers ending by several PLL/PGA-g-PEG bilayers can be used, PGA-g-PEG corresponding to PGA grafted by poly(ethylene glycol). Streaming potential and quartz crystal microbalance-dissipation measurements were used to characterize the buildup of these films. The multilayer films terminated by PGA and PGA-g-PEG were found to adsorb an extremely small amount of serum proteins as compared to a bare silica surface but the PGA ending films do not reduce bacterial adhesion. On the other hand, the adhesion of Escherichia coli bacteria is reduced by 72% on films ending by one (PLL/PGA-g-PEG) bilayer and by 92% for films ending by three (PLL/PGA-g-PEG) bilayers compared to bare substrate. Thus, our results show the ability of PGA-g-PEG to be inserted into multilayer films and to drastically reduce both protein adsorption and bacterial adhesion. This kind of anti-adhesive films represents a new and very simple method to coat any type of biomaterials for protection against bacterial adhesion and therefore limiting its pathological consequences. PMID- 14741615 TI - Effect of cobalt and chromium ions on bcl-2, bax, caspase-3, and caspase-8 expression in human U937 macrophages. AB - The bcl-2 and caspase families of proteins play a central role in the modulation of apoptosis. The purpose of this study was to analyze the effect of Co(2+) and Cr(3+) ions on the expression of bcl-2, bax, caspase-3 and caspase-8 to better understand the mechanisms leading to ion-induced apoptosis in macrophages. U937 human macrophages were exposed to Co(2+) and Cr(3+) ions. The expression of proteins was measured by Western blot while caspase activities were measured by colorimetric assay. Results show that Co(2+) ions inhibited bcl-2 expression with significant effect (p<0.05) after 16 h and a maximal 52% inhibitory effect after 24 h. Co(2+) stimulated bax expression with a significant stimulation (p<0.05) after 8 h and a maximal 1.75-fold increase after 16 h. Co(2+) also stimulated the expression of the active fragment of caspase-3 as well as caspase-3 activity maximal increase after 24 h. Co(2+) ions had no effect on caspase-8 expression or activity.Cr(3+) ions inhibited bcl-2 expression with significant effect (p<0.05) after 16 h and a maximal 43% inhibitory effect after 24 h. Cr(3+) stimulated bax expression with significant stimulation (p<0.01) after 8h and a maximal 2.25-fold increase after 24 h. Cr(3+) ions also stimulated the expression of the active fragments of caspase-3 and -8, as well as the activities of both proteases. The effect of Cr(3+) ions on the expression of both caspase active fragments was maximal after 16 h incubation. In conclusion, our results suggest that the modulation of the expression of proteins from the bcl-2 and the caspase families of proteins are implicated in the induction of macrophage apoptosis by Co(2+) and Cr(3+) ions. PMID- 14741616 TI - Electrostatic spray deposition (ESD) of calcium phosphate coatings, an in vitro study with osteoblast-like cells. AB - Electrostatic spray deposition (ESD) is a recently developed technique to deposit a calcium phosphate (CaP) coating upon substrates. With this technique, an organic solvent containing calcium and phosphate is pumped through a nozzle. Between the nozzle and substrate a high voltage is applied. As a consequence, droplets coming out the nozzle disperse into a spray, and this spray is deposited upon the substrate. When the solvent has evaporated, a coating is formed on the substrate. ESD allows for a variation in coating composition and morphology. Titanium alloy (TiAl6V4) substrates were coated with a CaP layer using two different methods; radio frequency magnetron sputtering, and ESD. These surfaces were characterized with X-ray diffraction, Fourier transform infrared spectroscopy, an universal surface tester, scanning electron microscopy, and energy dispersive spectrometry. Subsequently, bone marrow cells were isolated from rat femora and cultured 1, 4, 8, 14 and 16 days. Cell proliferation, alkaline phosphatase activity, and osteocalcin concentration were assayed. RT-PCR was done for collagen type I and osteocalcin. SEM was also performed to observe cellular behaviour during culture. Two separate runs of the experiment were performed. In the first run, osteoblast-like cells on both CaP coatings showed similar results in all assays. In the second run, proliferation and osteogenic expression had increased on ESD coatings. On basis of these results, we conclude that the novel ESD coating behaved similar to, or even better than the known RF magnetron sputter coating. Thus, ESD could be a valid addition to already existing CaP coating processes. PMID- 14741617 TI - Role of chemical interactions in bacterial adhesion to polymer surfaces. AB - Development of biomaterial-related infections is attracting an increasing interest due to the significant percentage of implant failure in the hospital care. Recent literature puts in evidence the dependence of the infection risk on the different biomaterials used, because of the different interactions between material surface and micro-organisms. Despite this, the mechanisms underlying the adhesion of bacteria to the biomaterial surface are still unclear. Aim of this work is to study the initial events of the processes responsible for the bacterial adhesion on polymers in order to prevent the development of bacterial infections and the consequent failure and replacement of biomedical devices. Electrostatic and Lifshitz-van der Waals forces are usually considered responsible for the interactions at the biomaterial interface. A new term that involves Lewis acid-base interactions is here introduced to better describe the bacterial adhesion to the polymer surface. Two requirements are needed to test this hypothesis: the development of an ideal polymeric surface in terms of chemical and morphological properties and the choice of a specific bacterial strain to be utilized as "probe". Experiments were worked out using an Escherichia coli (Gram-) strain that represent one of the principal isolates from infected biomaterial implants and its adhesion was investigated on polymers having different acid/basic character. The findings indicate that the bacterial adhesion is influenced by the chemical properties of the polymeric surface. These results may be interpreted taking into account a mechanism in which the acid/base (Lewis) interaction plays an important role. PMID- 14741618 TI - Nodule formation and mineralisation of human primary osteoblasts cultured on a porous bioactive glass scaffold. AB - The aim of this study was to analyse human osteoblast responses to a porous bioactive glass scaffold. It was hypothesised that osteoblasts would attach, proliferate and form mineralised nodules in response to culture on the bioactive glass. As dissolution products are a key feature of bioactive glasses, this was measured by inductively coupled plasma optical emission spectroscopy to determine effects of both the glass surface and ion release. Osteoblasts attached and proliferated on the foams as demonstrated by scanning electron microscopy. Nodule formation was also observed in the pores of the glass and also in conditioned medium containing dissolution products at certain concentrations and these nodules were shown to be mineralised by alizarin red staining. Undiluted dissolution products from the foams however caused significant apoptosis suggesting an ion concentration dependent response. PMID- 14741619 TI - Cell micropatterning using photopolymerization with a liquid crystal device commercial projector. AB - Photopolymerization has been widely used for surface micropatterning. The technique often requires photomasks and light sources with appropriate energies or filters. For rapid prototyping of surface photo-micropatterning, we have developed a novel device by modifying a commercially available liquid crystal device projector. In place of the image expansion unit of the projector, we attached an image reduction unit, an adjustable stage, and an optical monitoring unit. The device projected computer-generated images onto surfaces and subjected these patterns to photopolymerization. Micropatterned images can be easily prepared with various software run on personal computers. With the developed photopolymerization device, micropatterning of poly(ethylene glycol) (PEG) was achieved with PEG-diacrylate and a visible light photopolymerization initiator, camphorquinone. Selective cell adhesion control was also achieved on the micropatterned surfaces. PMID- 14741620 TI - Lithography application of a novel photoresist for patterning of cells. AB - Photolithography is the current workhorse for the microelectronic industry. It has been used extensively for the creation of patterns on two-dimensional surfaces. Various research groups have studied the use of photolithography to pattern surfaces for the alignment of cells. So far, these applications have been limited due to the use of organic solvents in the pattern developing process, which can denature biomacromolecules that would be attached to the material. To address this problem, a novel bioactive photoresist (bioresist) based on the copolymer of methyl methacrylate and 3-(t-butoxycarbonyl)-N-vinyl-2-pyrrolidone (MMA:TBNVP) was prepared and in vitro fibroblast cell growth on this resist was studied. Results demonstrated that the resist is non-adhesive to the fibroblast cells. By deprotecting the t-BOC groups into carboxyl groups (MMA:D-TBNVP), the material became cell adhesive. Furthermore, cells were able to proliferate on the MMA:D-TBNVP surface. By culturing cells on the MMA:D-TBNVP surface in serum versus serum-free medium, we reached the conclusion that the chemistry of the deprotected copolymer indirectly promoted cell attachment through its absorbance of serum proteins on the material. Patterns of 25 microm x 25 microm lines were obtained by chemically manipulating the surface of the photoresist using UV lithography without any solvent development. Fibroblast cells were observed to align on the patterned surface. This resist could be a suitable candidate to improve the application of conventional lithography in direct protein patterning for the guided growth of cells. PMID- 14741621 TI - Nano-fibrous poly(L-lactic acid) scaffolds with interconnected spherical macropores. AB - Biodegradable polymers have been used extensively as scaffolding materials to regenerate new tissues. These scaffolds should possess certain physical characteristics including a three-dimensional structure, high porosity with an interconnected pore structure, and a suitable surface structure for cell attachment, proliferation, and differentiation. To mimic the fibrous architecture of type I collagen, nano-fibrous matrices have been created in our laboratory using a phase-separation technique of poly(L-lactic acid) (PLLA) solutions. In addition, biodegradable scaffolds with controlled interconnected spherical pore networks have been fabricated in our laboratory. In this work, these two techniques were combined to yield scaffolds with highly interconnected spherical macroporous structures and nano-fibrous architectures. Paraffin spheres were first fabricated with a dispersion method, and were thermally bonded to form an interconnected mold. PLLA solutions were cast over the paraffin sphere assembly and were thermally phase-separated to form nano-fibrous matrices. After leaching out the paraffin, synthetic nano-fibrous extracellular matrices with interconnected spherical pores were yielded. By utilizing this fabrication process, we are able to control the architecture of the scaffolds at several different levels, including the macroscopic shape of the scaffold, the spherical pore size, interfiber distance, and the fiber diameter at the nano-size scale. The inter-pore connectivity could be controlled by varying the heat treatment time of the paraffin spheres, and mechanical properties could be controlled by varying the porosity of the scaffolds. With an interconnected macroporous structure that promotes cell seeding throughout the interstices of the scaffold, and a synthetic collagen-like matrix, these novel matrices may be an excellent scaffold for tissue engineering. PMID- 14741622 TI - Chitosan supports the initial attachment and spreading of osteoblasts preferentially over fibroblasts. AB - The aim of this study was to determine chitosan's effect on osteoblast and fibroblast cell attachment. Mouse MC3T3-E1 osteoblasts and 3T3 fibroblasts were grown in the presence of serum on two commercially available chitosans, Chitosan H (CH) and Protasan CL212 (PR). Cell attachment and immunofluorescent analysis at various time points were done to analyze initial phenotypic profiles. At 1h, significantly (P<0.05) fewer fibroblasts attached to CH or PR than serum-coated substrates. Osteoblast attachment to the same biopolymers at 1h was significantly greater than those seen with fibroblasts. At 24 h, levels of cell attachment for fibroblasts to both CH and PR significantly increased and were similar to levels seen in osteoblast cultures at both 1 and 24 h. Morphologically, immunofluorescent analysis showed that osteoblasts plated on the biopolymers were attached and beginning to spread at 1h, whereas fibroblasts appeared more rounded. At 24 h, fibroblasts plated on CH or PR revealed a heterogeneous population of round and semi-spread cells. In comparison, osteoblasts displayed phenotypes that were well spread with a developed cytoskeleton. These results suggest that CH and PR support the initial attachment and spreading of osteoblasts preferentially over fibroblasts, and that manipulation of the biopolymer can alter the level of attachment and spreading. PMID- 14741623 TI - Modulation of collagen proteolysis by chemical modification of amino acid side chains in acellularized arteries. AB - In this study, we have examined the effects of specific chemical modifications of amino acid side-chains on the in vitro degradation of "native" collagen obtained from acellular matrix (ACM)-processed arteries. Two monofunctional epoxides of different size and chemistry were used to modify lysine, with or without methylglyoxal modification of arginine. Biochemical, thermomechanical, tensile mechanical, and multi-enzymatic (collagenase, cathepsin B, acetyltrypsin, and trypsin) degradation analyses were used to determine the effects of modifications.Collagen solubilization by enzymes was found to depend upon the size and chemistry of epoxides used to modify lysine residues. In general, the solubilization of native ACM collagen by collagenase, cathepsin B, trypsin, and acetyltrypsin was either unaltered or decreased after modification with glycidol. In contrast, n-butylglycidylether (n-B) treatment increased solubilization by all enzymes. Subsequent arginine modification significantly reduced collagen solubilization by acetyltrypsin for glycidol-treated ACM arteries, whereas increased collagen solubilization was observed for n-B-treated ACM arteries with all enzymes. Gel chromatographic analyses of collagen fragments solubilized by trypsin revealed that both the amount and sites of cleavage were altered after lysine and arginine modification. The ability to modulate the enzymatic degradation of tissue-derived materials as demonstrated in this study may facilitate the design of novel engineering scaffolds for tissue regeneration or collagen-based drug delivery systems. PMID- 14741624 TI - Decreased fibroblast cell density on chemically degraded poly-lactic-co-glycolic acid, polyurethane, and polycaprolactone. AB - Select prolonged functions of fibroblasts leading to extensive fibrous tissue encapsulation can be detrimental to numerous implant applications, including materials designed for the bladder, vasculature, and bone. Specifically, overextended functions of fibroblasts at the tissue-implant interface for orthopedic applications lead to callus formation, fibrous encapsulation events, and ultimately soft (not desirable hard-bony) tissue juxtaposition. Such events result in insufficient regeneration of bone and compromise the overall success of the implant. The objective of the present in vitro study was to determine, for the first time, fibroblast densities on NaOH-treated poly-lactic-co-glycolic acid co-polymers (PLGA), HNO(3)-treated polyurethane (PU), and NaOH-treated polycaprolactone (PCL). Previous studies have demonstrated increased bladder, vascular, and bone cell densities on chemically treated compared to unaltered PLGA, PU, and PCL films. Results of this study provided evidence of decreased fibroblast numbers on chemically treated PLGA, PU, and PCL after time periods of up to 5 days. Examination of these substrates revealed that all chemically modified polymers possessed a high degree of nanometer surface roughness compared to their respective unaltered polymers. In contrast, other material properties (such as chemistry and wettability) were different when comparing chemically treated PLGA, PU, and PCL films. Since fibroblasts are anchorage-dependent cells whose adhesion is a critical prerequisite to the prolonged, extensive formation of a fibrous-tissue containing extracellular matrix, the present in vitro results of decreased fibroblast densities on chemically degraded PLGA, PU, and PCL suggest that these materials may be suitable materials for numerous tissue engineering applications and, thus, deserve further investigation. PMID- 14741625 TI - Effect of different sterilization processing methods on the mechanical properties of human cancellous bone allografts. AB - Use of new sterilization methods applied to human bone is likely to affect both the mechanical and biological properties of human cancellous grafts. The mechanical properties of the transplanted bone inevitably determine the short- and mid-term results of the orthopedic procedure performed. The aim of this study was to compare, under similar conditions, the mechanical effects of gamma irradiation, lipid extraction, and treatment with 6M urea on trabecular bone samples, through conventional mechanical tests and measurement of the ultrasound wave propagation rate. Deteriorations measured for gamma irradiation and lipid extraction were low: 2.4% and 2.5%, respectively, for ultrasound propagation wave measurements. They were clearly significant for protocol including 6M urea, corresponding to a loss of 30% in values measured in the control sample for the stress to failure, inciting prudence when grafted bone is used for support in orthopedic assembly. High consistency in the results obtained between travel time of the ultrasound wave, easily done, and measurement of stress to failure through conventional tests, favor the use of ultrasound protocol, described as a quality test performed on bone grafts in the tissue bank before distribution and implantation. PMID- 14741626 TI - Osteoblast response to hydroxyapatite doped with divalent and trivalent cations. AB - The present in vitro study doped hydroxyapatite (HA) with various metal cations (Mg(2+), Zn(2+), La(3+), Y(3+), In(3+), and Bi(3+)) in an attempt to enhance properties of HA pertinent to orthopedic and dental applications. X-ray diffraction material characterization indicated that the metal cations may have substituted for calcium in the HA crystal structure and that all of the doped HA formulations were single-phase and crystalline. Scanning electron microscopy analysis revealed a variety of grain sizes, depending on the dopant utilized. Energy-dispersive spectroscopy confirmed that the dopants added during synthesis were present and that all of the HA formulations synthesized were within the defined range of HA phase in the CaO-P(2)O(5)-H(2)O system. Lastly, Bi-doped HA had a slower dissolution rate than either undoped HA or HA doped with other cations when exposed to simulated physiological conditions for 21 days. In terms of cell function, results provided the first evidence that osteoblasts, bone forming cells, adhered and differentiated (as measured by alkaline phosphatase synthesis) in response to HA doped with trivalent cations (specifically, La(3+), Y(3+), In(3+), Bi(3+)) at earlier time points than either HA doped with divalent cations (Mg(2+), Zn(2+)) or undoped HA. Of the dopants examined, Bi(3+) most enhanced osteoblast long-term calcium-containing mineral deposition. For these reasons, this study revealed for the first time the potential benefits of doping HA with Bi(3+) according to criteria critical for bone prosthetic clinical success. PMID- 14741627 TI - Bone inductive properties of rhBMP-2 loaded porous calcium phosphate cement implants in cranial defects in rabbits. AB - In this study, the osteoinductive properties of porous calcium phosphate (Ca-P) cement loaded with bone morphogenetic protein 2 (rhBMP-2) were evaluated and compared with rhBMP-2 loaded absorbable collagen sponge (ACS). Discs with a diameter of 8mm were loaded with a buffer solution with or without 10 microg rhBMP-2 and inserted in 8mm full thickness cranial defects in rabbits for 2 and 10 weeks of implantation. Histological analysis revealed excellent osteoconductive properties of the Ca-P material. It maintained its shape and stability during the implantation time better than the ACS but showed no degradation like the ACS. Quantification of the Ca-P cement implants showed that bone formation was increased significantly by administration of rhBMP-2 (10 weeks pore fill: 53.0+/-5.4%), and also reached a reasonable amount without rhBMP-2 (43.1+/-10.4%). Remarkably, callus-like bone formation outside the implant was observed frequently in the 2 weeks rhBMP-2 loaded Ca-P cement implants, suggesting a correlation with the presence of growth factor in the surrounding tissue. However, an additional in vitro assay revealed an accumulative release of no more than 9.7+/-0.9% after 4 weeks. We conclude that: (1). Porous Ca-P cement is an appropriate candidate scaffold material for bone engineering. (2). Bone formation can be enhanced by lyophilization of rhBMP-2 on the cement. (3). Degradation of porous Ca-P cement is species-, implantation site- and implant dimension-specific. PMID- 14741628 TI - Surface immobilized bisphosphonate improves stainless-steel screw fixation in rats. AB - An increase in the mechanical fixation in bone of metallic biomaterials is considered advantageous in joint replacement and fracture surgery. Different approaches to improve fixation may be e.g. surface roughening, Ca-mineral coating or surface immobilization of growth factors or drugs. In the present work, bisphosphonate, a class of drugs that inhibit bone resorption, was immobilized onto stainless-steel screws. The screws were first roughened and coated with immobilized and cross-linked fibrinogen. Subsequently, an N-bisphosphonate, pamidronate, was immobilized onto fibrinogen, and another N-bisphosphonate, ibandronate, adsorbed on top of this. The so coated screws were inserted into the tibiae of eight male Sprague-Dawley rats. Another eight rats received screws prepared in the same way, but without the bisphosphonate coating. Pullout strength tests were performed after 2 weeks of implantation. The results showed a 28% (p=0.0009) higher pullout force and 90% increased pullout energy for the bisphosphonate coated screws, and support the idea that surface immobilized bisphosphonates can be used to improve biomaterials fixation in bone. PMID- 14741629 TI - Inhibition of bacterial adhesion on PVC endotracheal tubes by RF-oxygen glow discharge, sodium hydroxide and silver nitrate treatments. AB - Medical-grade poly(vinyl chloride) (PVC) was chemically modified to study how the incorporation of monovalent silver influences Pseudomonas aeruginosa adhesion and colonization. The modification investigated consisted of a radio frequency-oxygen (RF-O(2)) glow discharge pre-functionalization, followed by a two-step wet treatment in sodium hydroxide and silver nitrate solutions. X-ray photoelectron spectroscopy (XPS) analysis and contact angle measurements were used to investigate the chemical nature and surface wettability of the films following each step of the modification. XPS analysis proved that the RF-O(2) plasma pre functionalization of native PVC reproducibly increased the amount of functional groups representative of PVC additives, including ether/alcohol, esters and carboxyl groups. More specifically, we demonstrated that the O-C=O groups representative of the phthalic ester and zinc carboxylate additives identified for native PVC increased by two-fold following the RF-O(2) plasma pre functionalization step. Although RF-O(2) pre-functionalization did not have an effect on the silver content of the NaOH/AgNO(3) treated substrates, such a modification was necessary for biomaterial products that did not have reproducible surfaces amongst production lots. XPS analysis also demonstrated that saponification with sodium hydroxide (NaOH) of esters, like those of the phthalic ester additives of PVC is a simple, irreversible method of hydrolysis, which produced sodium carboxylate and sodium phthalate salts. Exposure of native PVC to NaOH resulted in an increased surface hydrophilicity (from ca 90 degrees to ca 60 degrees ) due to dechlorination. XPS analysis following further incubation in silver nitrate demonstrated that silver ions can be trapped when the sodium of sodium carboxylate is replaced by silver after performing a second treatment with a monovalent silver-containing solution. The creation of silver salt on native PVC resulted in an ultra-hydrophobic (>120 degrees ) surface. The chemical modifications using NaOH and AgNO(3) wet treatments completely inhibited bacterial adhesion of four strains of P. aeruginosa to both native and oxygen-pre functionalized PVC, and efficiently prevented colonization over longer periods (72 h). Our results suggest that surface modifications that incorporate silver ions would be extremely effective at reducing bacterial colonization to medical devices. PMID- 14741630 TI - Influence of grinding, sandblasting, polishing and heat treatment on the flexural strength of a glass-infiltrated alumina-reinforced dental ceramic. AB - The influence of processing-introduced flaws and heat treatment on the strength degradation of the dental core material has recently been observed. However, there are insufficient studies which investigate the role of grinding, grinding orientation, sandblasting, polishing and heat treatment on the strength of In Ceram Alumina (IA), one of the most used glass-infiltrated alumina-reinforced dental core ceramics. To address these issues, the uniaxial flexural strength and reliability of eight groups of specimens (sandblasted, ground parallel to the tensile axis, ground perpendicular, and polished with and without heat treatment) were assessed. Statistical analyses indicate that heat treatment significantly improved the flexural strength of the material regardless of the surface treatment. Conversely, any surface treatment caused strength degradation, if it was not followed by heat treatment. Sandblasting caused the most marked strength degradation. Polishing alone (without heat treatment) did not strengthen the ceramic. The orientation of grinding in respect of the direction of the tensile stresses did not influence the ultimate tensile strength. The present study suggests that, in the case of IA, sandblasting, grinding and polishing should always be followed by heat treatment in order to avoid strength degradation of the material. PMID- 14741631 TI - Macroporous poly(L-lactide) of controlled pore size derived from the annealing of co-continuous polystyrene/poly(L-lactide) blends. AB - A detailed study on the static annealing of co-continuous polystyrene/poly(L lactide) (PLLA) blends is presented. The effects of temperature, time at temperature, viscosity of the phases and interfacial modification on the coarsening of the blend are discussed. In this paper, polystyrene and PLLA are blended at compositions of 50/50 and 60/40 to form co-continuous morphologies. These co-continuous morphologies are coarsened under quiescent annealing conditions, and the subsequent removal of the polystyrene phase leaves a macroporous PLLA structure. The microstructure is analyzed using three different techniques: the BET nitrogen adsorption technique, mercury intrusion porosimetry and SEM combined with image analysis. It is shown that static annealing can be used to generate a series of co-continuous networks with controlled pore sizes ranging from 1 to hundreds of microns. A non-linear pore size growth rate is observed for these systems due to the degradation of PLLA and this study indicates that controlled degradation can be used as an additional tool for morphology control. Compatibilized polystyrene/PLLA blends demonstrate significantly reduced coarsening effects due to the reduction of interfacial tension. The coarsening rate of the co-continuous structure was examined in terms of the pore size, R and this growth rate is discussed in terms of a previously proposed coarsening mechanism. This approach is a route towards the preparation of a macroporous PLLA structure with pore sizes in the range required for scaffolds for tissue regeneration. PMID- 14741632 TI - An augmented hybrid constitutive model for simulation of unloading and cyclic loading behavior of conventional and highly crosslinked UHMWPE. AB - Ultra-high molecular weight polyethylene (UHMWPE) is extensively used in total joint replacements. Wear, fatigue, and fracture have limited the longevity of UHMWPE components. For this reason, significant effort has been directed towards understanding the failure and wear mechanisms of UHMWPE, both at a micro-scale and a macro-scale, within the context of joint replacements. We have previously developed, calibrated, and validated a constitutive model for predicting the loading response of conventional and highly crosslinked UHMWPE under multiaxial loading conditions (Biomaterials 24 (2003) 1365). However, to simulate in vivo changes to orthopedic components, accurate simulation of unloading behavior is of equal importance to the loading phase of the duty cycle. Consequently, in this study we have focused on understanding and predicting the mechanical response of UHMWPE during uniaxial unloading. Specifically, we have augmented our previously developed constitutive model to also allow for accurate predictions of the unloading behavior of conventional and highly crosslinked UHMWPE during cyclic loading. It is shown that our augmented hybrid model accurately captures the experimentally observed characteristics, including uniaxial cyclic loading, large strain tension, rate-effects, and multiaxial deformation histories. The augmented hybrid constitutive model will be used as a critical building block in future studies of fatigue, failure, and wear of UHMWPE. PMID- 14741633 TI - Comparative hardness and modulus of tooth-colored restoratives: a depth-sensing microindentation study. AB - The objective of this study was to compare the hardness and modulus of the continuum of direct tooth-colored restorative materials using a depth-sensing microindentation approach. The effects of thermal fatigue on mechanical properties were also evaluated. Six restorative materials representing the continuum were selected. They included an ormocer (Admira [AM], Voco), a giomer (Beautifil [BF], Shofu), a compomer (Dyract Extra [DE], Dentsply), a minifill composite (Esthet-X [EX], Dentsply), resin-modified (Fuji II LC [FL], GC) and highly viscous (Fuji IX [FN], GC) glass ionomer cements (GICs). Fourteen specimens (3 mm wide x 3 mm long x 2 mm deep) were made for each material. The specimens were randomly divided into two groups and treated as follows: Group A- stored in distilled water at 37 degrees C for 30 days; Group B--thermal cycled for 5000 cycles (35 degrees C [28s], 15 degrees C [2s], 35 degrees C [28s], 45 degrees C [2s]) and stored for 26.5 days. Hardness and modulus of the materials were determined using depth-sensing microindentation testing with the Instron MicroTester. Hardness was computed by dividing the peak load over the maximum projected contact area while modulus was calculated by analysis of the loading/unloading load-displacement (P-h) curves and the analytical model according to Oliver and Pharr (J. Mater. Res. 7 (1992) 1564). Results were analyzed using ANOVA/Scheffe's post hoc test and independent samples T-test (p<0.05). Hardness ranged 46.44-72.65 and 49.11-78.97 HV, while modulus ranged 7.86-12.78 and 8.12-13.13 GPa for Groups A and B, respectively. Although the ranking of mechanical properties were generally similar for both groups, disparities in statistical differences between materials were observed between Groups A and B for both hardness and modulus. For both groups, BF was significantly harder than DE, AM, FL and EX was significantly harder than FL. The modulus of FN was significantly greater than EX, DE, AM and FL was significantly stiffer than AM. With the exception of BF, no significant change in hardness and modulus was observed for all materials with thermocycling. The hardness and modulus of some glass ionomer-based/containing materials may be comparable or even superior to minifill and ormocer composites. Thermal fatigue should be considered when comparing mechanical properties between materials. PMID- 14741634 TI - Ionic modification of calcium phosphate cement viscosity. Part I: hypodermic injection and strength improvement of apatite cement. AB - A broadening of the indications for which calcium phosphate cements (CPC) can be used, for example, in the field of vertebroplasty, would require injectable and higher strength materials. Unmodified CPC are not injectable due to a filter pressing effect during injection. In this work we demonstrated that an effective method for improving the injection properties of CPC was by the use of sodium citrate solution as a liquid component. Cement consisting of tetracalcium phosphate (TTCP) and monetite (DCPA) mixed with water up to a powder:liquid ratio (P:L) of 3.3 g/ml had an injectability of approximately 60%. The use of 500 mM trisodium citrate solution instead of water decreased the viscosity of the cement paste to a point, where complete injectability (>95%) through an 800 microm diameter hypodermic needle could be achieved at low loads. The reduction in water demand of the cement effected by the use of sodium citrate enabled high P:L mixes to be formed which were 400% stronger than cements made with water. The effect was less pronounced with compacted cements such that at 9 MPa applied pressure, 58% improvement was obtained and at 50 MPa 36% improvement was measured yielding a cement with a compressive strength of 154 MPa. The liquefying effect of sodium citrate was thought to derive from a strong increase in the surface charge of both the reactants and the product as determined by zeta-potential measurement. PMID- 14741635 TI - Ionic modification of calcium phosphate cement viscosity. Part II: hypodermic injection and strength improvement of brushite cement. AB - Brushite-forming calcium phosphate cements are of great interest as bone replacement materials because they are resorbable in physiological conditions. However, their short setting times, low mechanical strengths and limited injectability limit broad clinical application. In this study, we showed that a significant improvement of these properties of brushite cement could be achieved by the use of sodium citrate or citric acid as setting retardants, such that workable cement pastes with a powder to liquid ratio of up to 5 could be manufactured. The cement used in this study consisted of an equimolar powder mixture of beta-tricalcium phosphate and monocalcium phosphate hydrate The use of 500 mM-1M retardant solutions as liquid phase enabled initial setting times of 8 12 min. Wet compressive strength were found to be in the range between 12-18 MPa after immersion of uncompacted cement samples in serum for 24 h. A further strength improvement to 32 MPa was obtained by compaction of the cement paste during samples preparation. This is significant because high-temperature processes cannot be used to fabricate hydrated calcium phosphate materials. Cement pastes were injectable through a hypodermic needle at a powder to liquid ratio of 3.3 g/ml when a 1M citric acid was used as liquid phase, thus enabling precise controlled delivery to small defects. PMID- 14741636 TI - Rietveld structure refinement of precipitated carbonate apatite using neutron diffraction data. AB - X-ray and time of flight neutron diffraction data, FTIR and MAS-NMR spectra, and ICP-AES and carbonate analyses have been collected from a sodium-containing carbonate apatite (CO(3) content 12.5(7)wt%). A structural model based on Holly Springs hydroxyapatite without CO(3)(2-) ions showed an apparent reduction in the PO(4) tetrahedral volume which is ascribed to CO(3)(2-) replacing PO(4)(3-) ions in the lattice. Four structural models from the literature with the CO(3)(2-) ion explicitly modelled were fitted to the neutron diffraction data by the Rietveld method. The best fit was obtained with the CO(3)(2-) ion in disorder between the mirror symmetry related faces of a vacant PO(4)(3-) site and with the normal to the plane of the CO(3)(2-) ion at approximately 30 degrees to the c-axis. This angle is consistent with results from previous polarised IR measurements on single crystals of francolite (a fluorocarbonate apatite) and human dental enamel. Further refinement of the model revealed a hitherto unknown atom site close to the unit cell origin, (possibly a water molecule). The refined hexagonal unit cell parameters from the neutron diffraction data were a=9.3446(3)A and c=6.9199(4)A. PMID- 14741637 TI - Functional roles of the cingulo-frontal network in performance on working memory. AB - We examined the relationship between brain activities and task performance on working memory. A large-scale study was initially administered to identify good and poor performers using the operation span and reading span tasks. On the basis of those span scores, we divided 20 consenting participants into high- and low span groups. In an fMRI study, the participants performed verification of arithmetic problems and retention of target words either concurrently or separately. The behavioral results showed that performance was better in the high span group than in the low-span group under a dual-task condition, but not under two single-task conditions. The anterior cingulate cortex (ACC), left prefrontal cortex (PFC), left inferior frontal cortex, and bilateral parietal cortex were primarily activated for both span groups. We found that signal changes in the ACC were greater in the high-span group than in the low-span group under the dual task condition, but not under the single-task conditions. Structural equation modeling indicated that an estimate of effective connectivity from the ACC to the left PFC was positive for the high-span group and negative for the-low span group, suggesting that closer cooperation between the two brain regions was strongly related to working memory performance. We conclude that central executive functioning for attention shifting is modulated by the cingulo-frontal network. PMID- 14741638 TI - A myelo-architectonic method for the structural classification of cortical areas. AB - We describe an automatic and reproducible method to analyze the histological design of the cerebral cortex as applied to brain sections stained to reveal myelinated fibers. The technique provides an evaluation of the distribution of myelination across the width of the cortical mantle in accordance with a model of its curvature and its intrinsic geometry. The profile lines along which the density of staining is measured are generated from the solution of a partial differential equation (PDE) that models the intermediate layers of the cortex. Cortical profiles are classified according to significant components that emerge from wavelet analysis. Intensity profiles belonging to each distinct class are normalized and averaged to produce area-specific templates of cortical myelo architecture. PMID- 14741639 TI - Focal white matter density changes in schizophrenia: reduced inter-hemispheric connectivity. AB - Gray matter changes have been demonstrated in several regions in schizophrenia. Particularly, the frontal and temporal cortices and amygdala-hippocampal region have been found decreased in volume and density in magnetic resonance imaging (MRI) studies. These abnormalities may reflect an aberrant neuronal network in schizophrenia, suggesting that white matter fibers connecting these regions may also be affected. However, it is unclear if particular white matter areas are (progressively) affected in schizophrenia and if these are related to the gray matter changes. Focal white matter changes in schizophrenia were studied in whole brain magnetic resonance images acquired from 159 patients with schizophrenia or schizophreniform disorder and 158 healthy comparison subjects using voxel-based morphometry. White matter density changes in the patients with schizophrenia were correlated to gray matter density changes and to illness severity. In the patients with schizophrenia, significant decreases in white matter density were found in the genu and truncus of the corpus callosum in the left and right hemisphere, in the right anterior internal capsule and in the right anterior commissure. No interactions between diagnosis and age were found. Increased illness severity was correlated with low density of the corpus callosum and anterior commissure. Decreased corpus callosum density correlated with decreased density of thalamus, lateral inferior frontal and insular gray matter in patients and controls and with decreased density of medial orbitofrontal and superior temporal gyri in patients. Decreased internal capsule and anterior commissure density correlated with increased caudate, and globus pallidus density in patients and controls. These findings suggest aberrant inter-hemispheric connectivity of anterior cortical and sub-cortical brain regions in schizophrenia, reflecting decreased hemispheric specialisation in schizophrenia. PMID- 14741640 TI - Feature-based attention in human visual cortex: simulation of fMRI data. AB - We use a computational neuroscience approach to study the role of feature-based attention in visual perception. This model is used to numerically simulate a visual attention experiment. The neurodynamical system consists of many interconnected modules that can be related to the dorsal and ventral paths of the visual cortex. The biased competition hypothesis is taken into account within the model. From the experimental point of view, measurements exist, which confirm that feature-based attention influences visual cortical responses to stimuli outside the attended location. These measurements show that attention to a given stimulus attribute (in this case "color red") increases the response of cortical visual areas to a spatially distant, ignored stimulus that shares the same attribute. Our neurodynamical model is used to numerically compute the neural activity of area V4 corresponding to such ignored stimulus, giving a good description of the experimental data. PMID- 14741641 TI - Morphological classification of brains via high-dimensional shape transformations and machine learning methods. AB - A high-dimensional shape transformation posed in a mass-preserving framework is used as a morphological signature of a brain image. Population differences with complex spatial patterns are then determined by applying a nonlinear support vector machine (SVM) pattern classification method to the morphological signatures. Significant reduction of the dimensionality of the morphological signatures is achieved via wavelet decomposition and feature reduction methods. Applying the method to MR images with simulated atrophy shows that the method can correctly detect subtle and spatially complex atrophy, even when the simulated atrophy represents only a 5% variation from the original image. Applying this method to actual MR images shows that brains can be correctly determined to be male or female with a successful classification rate of 97%, using the leave-one out method. This proposed method also shows a high classification rate for old adults' age classification, even under difficult test scenarios. The main characteristic of the proposed methodology is that, by applying multivariate pattern classification methods, it can detect subtle and spatially complex patterns of morphological group differences which are often not detectable by voxel-based morphometric methods, because these methods analyze morphological measurements voxel-by-voxel and do not consider the entirety of the data simultaneously. PMID- 14741642 TI - Task demand modulations of visuospatial processing measured with functional magnetic resonance imaging. AB - Brain imaging based on functional magnetic resonance imaging (fMRI) provides a useful tool to examine neural networks and cerebral structures subserving visuospatial function. It allows not only the qualitative determination of which areas are active during task processing, but also estimates the quantitative contribution of involved brain regions to different aspects of spatial processing. In this study, we investigated in 10 healthy subjects how the amount of task (computational) demand in an angle discrimination task was related to neural activity as measured with event-related fMRI. Task demand, indicated by behavioral performance, was modulated by presenting clocks with different angular disparity and length of hands. Significant activations were found in the cortical network subserving the visual and visuospatial processing, including the right and left superior parietal lobules (SPL), striate visual areas, and sensorimotor areas. Both blood oxygenation level-dependent (BOLD) signal strength and spatial extent of activation in right as well as left SPL increased with task demand. By contrast, no significant correlation or a very weak correlation was found between the task demand and the BOLD signal as well as between task demand and spatial extent of activations in the striate visual areas and in the sensorimotor areas. These results support the hypothesis that increased computational demand requires more brain resources. The brain regions that are most specialized for the execution of the visuospatial task can be assessed by relating the imposed task demand to the functional activation measured. PMID- 14741643 TI - Neural correlates of auditory sensory memory and automatic change detection. AB - An auditory event-related potential component, the mismatch negativity (MMN), reflects automatic change detection and its prerequisite, sensory memory. This study examined the neural correlates of automatic change detection using BOLD fMRI and two rates of presentation previously shown to induce either a large or no MMN. A boxcar block design was employed in two functional scans, each performed twice. A block consisting of 1000-Hz standards (S) alternated with one consisting of 1000-Hz standards and 2000-Hz infrequent deviants (S + D). Presentation rate was either 150 or 2400 ms. Fourteen participants were instructed to ignore all auditory stimulation and concentrate on a film (no audio) by reading subtitles. Data analysis used SPM99 and random effects approach. Cluster statistics (P < 0.05, corrected) were employed at a height threshold of P < 0.001. At the short ISI, there was a significant BOLD response in the right superior temporal gyrus (STG), the left insula, and the left STG (including parts of primary auditory cortex). There were no suprathreshold clusters at the long rate, with S + D blocks inducing no greater activity than S blocks. These results support the hypothesis that the automatic detection of auditory change occurs in the STG bilaterally and relies on the maintenance of sensory memory traces. PMID- 14741644 TI - Dissociation between overt and unconscious face processing in fusiform face area. AB - The precise role of the fusiform face area (FFA) in face processing remains controversial. In this study, we investigated to what degree FFA activation reflects additional functions beyond face perception. Seven volunteers underwent rapid event-related functional magnetic resonance imaging while they performed a face-encoding and a face-recognition task. During face encoding, activity in the FFA for individual faces predicted whether the individual face was subsequently remembered or forgotten. However, during face recognition, no difference in FFA activity between consciously remembered and forgotten faces was observed, but the activity of FFA differentiated if a face had been seen previously or not. This demonstrated a dissociation between overt recognition and unconscious discrimination of stimuli, suggesting that physiological processes of face recognition can take place, even if not all of its operations are made available to consciousness. PMID- 14741645 TI - Pausing for thought: engagement of left temporal cortex during pauses in speech. AB - Pauses during continuous speech, particularly those that occur within clauses, are thought to reflect the planning of forthcoming verbal output. We used functional Magnetic Resonance Imaging (fMRI) to examine their neural correlates. Six volunteers were scanned while describing seven Rorschach inkblots, producing 3 min of speech per inkblot. In an event-related design, the level of blood oxygenation level dependent (BOLD) contrast during brief speech pauses (mean duration 1.3 s, SD 0.3 s) during overt speech was contrasted with that during intervening periods of articulation. We then examined activity associated with pauses that occurred within clauses and pauses that occurred between grammatical junctions. Relative to articulation during speech, pauses were associated with activation in the banks of the left superior temporal sulcus (BA 39/22), at the temporoparietal junction. Continuous speech was associated with greater activation bilaterally in the inferior frontal (BA 44/45), middle frontal (BA 8) and anterior cingulate (BA 24) gyri, the middle temporal sulcus (BA 21/22), the occipital cortex and the cerebellum. Left temporal activation was evident during pauses that occurred within clauses but not during pauses at grammatical junctions. In summary, articulation during continuous speech involved frontal, temporal and cerebellar areas, while pausing was associated with activity in the left temporal cortex, especially when this occurred within a clause. The latter finding is consistent with evidence that within-clause pauses are a correlate of speech planning and in particular lexical retrieval. PMID- 14741646 TI - Neural responses to Mooney images reveal a modular representation of faces in human visual cortex. AB - The way in which information about objects is represented in visual cortex remains controversial. One model of human object recognition poses that information is processed in modules, highly specialised for different categories of objects; an opposing model appeals to a distributed representation across a large network of visual areas. We addressed this debate by monitoring activity in face- and object-selective areas while human subjects viewed ambiguous face stimuli (Mooney faces). The measured neural response in the face-selective region of the fusiform gyrus was greater when subjects reported seeing a face than when they perceived the image as a collection of blobs. In contrast, there was no difference in magnetic resonance response between face and no-face perceived events in either the face-selective voxels of the superior temporal sulcus or the object-selective voxels of the parahippocampal gyrus and lateral occipital complex. These results challenge the concept that neural representation of faces is distributed and overlapping and suggest that the fusiform gyrus is tightly linked to the awareness of faces. PMID- 14741647 TI - Three-dimensional probabilistic anatomical cranio-cerebral correlation via the international 10-20 system oriented for transcranial functional brain mapping. AB - The recent advent of multichannel near-infrared spectroscopy (NIRS) has expanded its technical potential for human brain mapping. However, NIRS measurement has a technical drawback in that it measures cortical activities from the head surface without anatomical information of the object to be measured. This problem is also found in transcranial magnetic stimulation (TMS) that transcranially activates or inactivates the cortical surface. To overcome this drawback, we examined cranio cerebral correlation using magnetic resonance imaging (MRI) via the guidance of the international 10-20 system for electrode placement, which had originally been developed for electroencephalography. We projected the 10-20 standard cranial positions over the cerebral cortical surface. After examining the cranio-cerebral correspondence for 17 healthy adults, we normalized the 10-20 cortical projection points of the subjects to the standard Montreal Neurological Institute (MNI) and Talairach stereotactic coordinates and obtained their probabilistic distributions. We also expressed the anatomical structures for the 10-20 cortical projection points probabilistically. Next, we examined the distance between the cortical surface and the head surface along the scalp and created a cortical surface depth map. We found that the locations of 10-20 cortical projection points in the standard MNI or Talairach space could be estimated with an average standard deviation of 8 mm. This study provided an initial step toward establishing a three-dimensional probabilistic anatomical platform that enables intra- and intermodal comparisons of NIRS and TMS brain imaging data. PMID- 14741648 TI - Imagery in sentence comprehension: an fMRI study. AB - This study examined brain activation while participants read or listened to high imagery sentences like The number eight when rotated 90 degrees looks like a pair of spectacles or low-imagery sentences, and judged them as true or false. The sentence imagery manipulation affected the activation in regions (particularly, the intraparietal sulcus) that activate in other mental imagery tasks, such as mental rotation. Both the auditory and visual presentation experiments indicated activation of the intraparietal sulcus area in the high-imagery condition, suggesting a common neural substrate for language-evoked imagery that is independent of the input modality. In addition to exhibiting greater activation levels during the processing of high-imagery sentences, the left intraparietal sulcus also showed greater functional connectivity in this condition with other cortical regions, particularly language processing regions, regardless of the input modality. The comprehension of abstract, nonimaginal information in low imagery sentences was accompanied by additional activation in regions in the left superior and middle temporal areas associated with the retrieval and processing of semantic and world knowledge. In addition to exhibiting greater activation levels during the processing of low-imagery sentences, this left temporal region also revealed greater functional connectivity in this condition with other left hemisphere language processing regions and with prefrontal regions, regardless of the input modality. The findings indicate that sentence comprehension can activate additional cortical regions that process information that is not specifically linguistic but varies with the information content of the sentence (such as visual or abstract information). In particular, the left intraparietal sulcus area appears to be centrally involved in processing the visual imagery that a sentence can evoke, while activating in synchrony with some core language processing regions. PMID- 14741649 TI - Rapid discrimination of visual and multisensory memories revealed by electrical neuroimaging. AB - Though commonly held that multisensory experiences enrich our memories and that memories influence ongoing sensory processes, their neural mechanisms remain unresolved. Here, electrical neuroimaging shows that auditory-visual multisensory experiences alter subsequent processing of unisensory visual stimuli during the same block of trials at early stages poststimulus onset and within visual object recognition areas. We show this with a stepwise analysis of scalp-recorded event related potentials (ERPs) that statistically tested (1) ERP morphology and amplitude, (2) global electric field power, (3) topographic stability of and changes in the electric field configuration, and (4) intracranial distributed linear source estimations. Subjects performed a continuous recognition task, discriminating repeated vs. initial image presentations. Corresponding, but task irrelevant, sounds accompanied half of the initial presentations during a given block of trials. On repeated presentations within a block of trials, only images appeared, yielding two situations-the image's prior presentation was only visual or with a sound. Image repetitions that had been accompanied by sounds yielded improved memory performance accuracy (old or new discrimination) and were differentiated as early as approximately 60-136 ms from images that had not been accompanied by sounds through generator changes in areas of the right lateral occipital complex (LOC). It thus appears that unisensory percepts trigger multisensory representations associated with them. The collective data support the hypothesis that perceptual or memory traces for multisensory auditory-visual events involve a distinct cortical network that is rapidly activated by subsequent repetition of just the unisensory visual component. PMID- 14741650 TI - In vivo study of acetylcholine esterase in basal forebrain, amygdala, and cortex in mild to moderate Alzheimer disease. AB - It is currently unclear whether impairment of the cholinergic system is present in Alzheimer disease (AD) already at an early stage and to what extent it depends on degeneration of the nucleus basalis of Meynert (nbM). We examined acetylcholine esterase activity in vivo in the nbM, the amygdala, and cerebral neocortex. Measurements were performed in normal controls and in patients with mild to moderate AD with positron emission tomography (PET) and C-11-labeled N methyl-4-piperidyl-acetate (MP4A) which is a specific substrate of AChE. AChE activity was reduced significantly in amygdala and cerebral cortex. In contrast, AChE activity and glucose metabolism appeared preserved or even increased in the nbM. The results support the concept that neocortical and amygdaloid functional changes of the cholinergic system are an early and leading event in AD, rather than the consequence of neurodegeneration of basal nuclei. PMID- 14741651 TI - Discrepancies between BOLD and flow dynamics in primary and supplementary motor areas: application of the balloon model to the interpretation of BOLD transients. AB - The blood-oxygen-level-dependent (BOLD) signal measured in the brain with functional magnetic resonance imaging (fMRI) during an activation experiment often exhibits pronounced transients at the beginning and end of the stimulus. Such transients could be a reflection of transients in the underlying neural activity, or they could result from transients in cerebral blood flow (CBF), cerebral metabolic rate of oxygen (CMRO2), or cerebral blood volume (CBV). These transients were investigated using an arterial spin labeling (ASL) method that allows simultaneous measurements of BOLD and CBF responses. Responses to a finger tapping task (40-s stimulus, 80-s rest) were measured in primary motor area (M1) and supplementary motor area (SMA) in five healthy volunteers. In SMA, the average BOLD response was pronounced near the beginning and end of the stimulus, while in M1, the BOLD response was nearly flat. However, CBF responses in the two regions were rather similar, and did not exhibit the same transient features as the BOLD response in SMA. Because this suggests a hemodynamic rather than a neural origin for the transients of the BOLD response in SMA, we used a generalization of the balloon model to test the degree of hemodynamic transients required to produce the measured curves. Both data sets could be approximated with modest differences in the shapes of the CMRO2 and CBV responses. This study illustrates the utility and the limitations of using theoretical models combined with ASL techniques to understand the dynamics of the BOLD response. PMID- 14741652 TI - Discrimination of temporal information at the cerebellum: functional magnetic resonance imaging of nonverbal auditory memory. AB - Until recently, the cerebellum was held to play its chief role in motor control. By contrast, Keele and Ivry (1990) proposed that it may subserve time estimation within the perceptual domain as well. In accordance with this suggestion, speech perception requiring minute differentiation of time intervals was found compromised by cerebellar pathology a subsequent functional magnetic resonance imaging (fMRI) study found hemodynamic activation of the right neocerebellum under these conditions. In the current fMRI investigation a non-speech task involving duration storage and comparison yielded significant hemodynamic responses within the lateral Crus I area of the right cerebellar hemisphere. Concomitantly, a left prefrontal cluster was observed. The present fMRI study employed single-shot double-echo echo-planar imaging (EPI) to reduce image distortion and acquisition time with whole-brain coverage (TE = 28 and 66 ms, TR = 5 s, 28 slices, TA = 2.8 s). Twelve healthy subjects performed two tasks: identifying pauses between tones as "short" or "long" (30-130 ms) and deciding which of two successive pauses was longer. The activation pattern in the discrimination task was analogous to that seen during speech perception and verbal working memory (WM) tasks. We suggest that the storage of precise temporal structures relies on a cerebellar-prefrontal loop. This network allows for temporal organization of verbal sequences and phoneme encoding based on durational operations in a linguistic context. PMID- 14741653 TI - MRS shows syndrome differentiated metabolite changes in human-generalized epilepsies. AB - OBJECTIVE: While it is generally accepted that the thalamo-cortical loop is abnormal in idiopathic generalized epilepsy (IGE), it is uncertain whether this loop is similarly affected among different IGE syndromes. We recently demonstrated reduced frontal lobe levels of N-acetyl aspartate (NAA) in patients with juvenile myoclonic epilepsy (JME). The present follow-up study investigates if similar or other types of changes exist in subjects with pure primarily generalized tonic clonic epilepsy (GTCS). METHOD: Twenty patients with GTCS, 26 patients with JME, and 10 matched healthy controls were investigated with quantitative single voxel MR spectroscopy (MRS) measurements of NAA, choline (Cho), creatine (Cr), and myo-inositol (mI) at 1.5 T scanner. The voxels were placed over the right cerebellum, right thalamus, prefrontal, occipital cortex, and over a spherical phantom above the subject's head. RESULTS: Patients with JME had reduced frontal lobe NAA (mmol/l) in relation to controls (9.8 +/- 1.1 vs. 10.8 +/- 0.7, P = 0.01), as well as GTCS patients (9.8 +/- 1.1 vs. 10.6 +/- 0.7, P = 0.007), whose values were normal. Patients with GTCS, on the other hand, showed significantly lower thalamic NAA than controls (9.7 +/- 1.0 vs. 10.8 +/- 0.9, P = 0.002), and both groups of patients had reduced thalamic Cho, and mI; [CHO: 2.0 +/- 0.4 (control) vs. 1.61 +/- 0.3 (JME) P = 0.001, and vs. 1.57 +/- 0.3 (GTCS) P = 0.0005; MI: 4.8 +/- 1.5 (control) vs. 3.3 +/- 1.4 (JME) P = 0.003, and vs. 3.2 +/- 1.5 (GTCS), P = 0.002]. No other regional changes were observed. CONCLUSION: The present MRS data emphasize the involvement of thalamus in IGE. They also show partly differentiated alterations within the thalamo-cortical loop in JME vs. GTCS. The various clinical expressions of IGE may, thus, be associated with more localized neuroanatomical substrates than generally believed. PMID- 14741654 TI - Interindividual differences of medial temporal lobe activation during encoding in an elderly population studied by fMRI. AB - Functional MRI (fMRI) is used to study medial temporal lobe (MTL) activation during encoding of new information into memory. In most studies, fMRI data of different subjects are averaged in standard coordinate space. However, interindividual differences in activation can be extensive, reflecting functional heterogeneity. Further, anatomical differences in brain structure cause additional variance and loss of registration accuracy. Such differences in structural and functional MTL characteristics may interfere with the efficiency of averaging data across subjects, and may become more significant with aging and dementia. The current study concerns the analysis of individual differences in MTL activation associated with episodic encoding.Twenty-nine healthy elderly men between 60 and 70 years old performed a simple face encoding task during fMRI scanning. Individual data were analyzed in native space, and compared to the group average in standard space (Talairach and Tournoux).MTL volumes between subjects varied between 6.34 and 11.27 cm(3), and had considerable variation when mapped to standard space. Eighteen of the 29 subjects showed MTL activity and activation patterns varied both in location and size (ranging from 0.11 to 1.78 cm(3)), with the strongest activation in the left posterior part of the MTL. In standard space, no region was significantly activated on a group level at a comparable alpha level. We conclude that while the majority of elderly subjects show MTL activation during episodic encoding of faces, there is considerable structural and functional variability between subjects. Group analysis in standard space may not be appropriate for studies of a complex structure such as the MTL, particularly not in aging and dementia. PMID- 14741655 TI - Behavioral and functional MRI study of attention shift in human verbal working memory. AB - The tripartite model of memory proposed the requirement of attentional switching when accessing different items in working memory [J. Exp. Psychol. Learn. Mem. Cogn. 27 (2001) 817]. This internal focus of attention is limited to just one item and the switching process is time-consuming [Mem. Cogn. 26 (1998) 263]. In the current study, given a three-digit list stored in working memory, we found that it took longer to shift attention in the direction of "Upstream" than "Downstream", and that each shift was a "single step" process. To investigate the neural basis of this type of attention switching, we performed a functional MRI study. The results revealed that at least three important brain areas are involved, including the left dorsal lateral prefrontal cortex, the cingulate gyrus, and the medial occipital cortex. These areas all showed greater activation in the attention shift condition compared to control conditions of no (or decreased) attention shift requirements. In addition, the hemodynamic activities in these areas are highly correlated, suggesting a strong functional connectivity between them. Taken together with evidence from several recent investigations, our results suggest that these areas each play an important and specific role in collaboratively supporting the function of attention shift in working memory. PMID- 14741656 TI - fMRI of healthy older adults during Stroop interference. AB - The Stroop interference effect, caused by difficulty inhibiting overlearned word reading, is often more pronounced in older adults. This has been proposed to be due to declines in inhibitory control and frontal lobe functions with aging. Initial neuroimaging studies of inhibitory control show that older adults have enhanced activation in multiple frontal areas, particularly in inferior frontal gyrus, indicative of recruitment to aid with performance of the task. The current study compared 13 younger and 13 older adults, all healthy and well educated, who completed a Stroop test during functional magnetic resonance imaging. Younger adults were more accurate across conditions, and both groups were slower and less accurate during the interference condition. The groups exhibited comparable activation regions, but older adults exhibited greater activation in numerous frontal areas, including the left inferior frontal gyrus. The results support the recruitment construct and suggest, along with previous research, that the inferior frontal gyrus is important for successful inhibition. PMID- 14741657 TI - Technical issues in the determination of cerebrovascular reserve in elderly subjects using 15O-water PET imaging. AB - The accurate determination of cerebrovascular reserve (CVR), especially in elderly subjects, entails several technical issues. From a review of the literature, the optimal technique employs quantitative 15O-water PET imaging determinations of cerebral blood flow (CBF) and acetazolamide (ACZ) (1 g iv with measurements at 10- to 20-min post-administration) as the vasodilating agent. CBF and CVR measurements were made using this methodology on 12 elderly subjects (3 males, 9 females, 66-84 years of age) meeting criteria for mild cognitive impairment (MCI) without other significant medical problems. Applying this quantitative technique, the cognitive and emotional status of the subject during the imaging procedure influenced the magnitude of the measurements. The semiquantitative measures resulted in even more pronounced subject state influences. The conditions under which CBF or CVR measurements are made should be controlled and reported. If semiquantitative techniques (e.g., single-photon emission-computed tomography [SPECT] imaging) must be employed for the determination of CVR, the validity of any measurement is dependent on the careful control of the general physiological status (e.g., heart rate, blood pressure, level of anxiety) of the patient. PMID- 14741658 TI - Dichotic listening reveals functional specificity in prefrontal cortex: an fMRI study. AB - The present study used fMRI to investigate the relationship between stimulus presentation mode and attentional instruction in a free-report dichotic listening (DL) task with consonant-vowel (CV) syllables. Binaural and dichotic CV syllables were randomly presented to the subjects during four different instructional conditions: a passive listening instruction and three active instructions where subjects listened to both ears, right ear and left ear, respectively. The results showed that dichotic presentations activated areas in the superior temporal gyrus, middle and inferior frontal gyrus and the cingulate cortex to a larger extent than binaural presentations. Moreover, the results showed that increase of activation in these areas was differentially dependent on presentation mode and attentional instruction. Thus, it seems that speech perception, as studied with the DL procedure, involves a cortical network extending beyond primary speech perception areas in the brain, also including prefrontal cortex. PMID- 14741659 TI - Artifactual fMRI group and condition differences driven by performance confounds. AB - Analysis techniques comparing groups or conditions that vary in performance are open to a possible confound driven by those performance differences, if these errors are ignored. Disproportionate numbers of errors may either introduce noise into the signal of interest or confound the signal of interest with additional signal associated with specific error-related processes. Two inhibitory task datasets were reanalysed, one comparing young and elderly groups, the other comparing high and low conflict conditions within the same group of subjects. The data were analysed twice using event-related techniques, one treating correct and error responses separately, the other treating error responses as if they were correct. It was found that the activation maps differed considerably, with the inclusion of errors leading to many false positive and false negative activation clusters. Using performance as a covariate, analyses of covariance (ANCOVA) were used to try to correct these differences without success. Data simulations that varied the number of errors included in the analyses found that surprisingly few errors could significantly alter activation maps. Consequently, brain-imaging investigations that do not accommodate error contributions to functional signals are at risk of misinterpreting activation patterns. PMID- 14741660 TI - Differentiation of idiopathic Parkinson's disease, multiple system atrophy, progressive supranuclear palsy, and healthy controls using magnetization transfer imaging. AB - The differentiation of multiple system atrophy (MSA) and progressive supranuclear palsy (PSP) from idiopathic Parkinson's disease (IPD) is difficult. Magnetization transfer imaging (MTI), a measure that correlates with myelination and axonal density, was employed in this study in the attempt to distinguish between these disorders. Measurements were carried out in 15 patients with IPD, 12 patients with MSA, 10 patients with PSP, and in 20 aged-matched healthy control subjects. The main finding was a change in the magnetization transfer ratio in the globus pallidus, putamen, caudate nucleus, substantia nigra, and white matter in IPD, MSA, and PSP patients, matching the pathological features of the underlying disorder. Furthermore, stepwise linear discriminant analysis provided a good classification of the individual patients into the different disease groups. All IPD patients and control subjects were correctly separated from the MSA and PSP cohort, and all PSP patients and 11 of 12 MSA patients were correctly separated from the IPD and control cohort. There was also a fairly good discrimination of IPD patients from control subjects and of MSA from PSP patients. In conclusion, MTI revealed degenerative changes in patients with different parkinsonian syndromes matching the underlying pathological features of the different diseases, underlining the high potential of this method in distinguishing MSA and PSP from IPD. PMID- 14741661 TI - Reduced BOLD response to periodic visual stimulation. AB - The blood oxygenation level-dependent (BOLD) response to entrained neuronal firing in the human visual cortex and lateral geniculate nuclei was investigated. Periodic checkerboard flashes at a range of frequencies (4-20 Hz) were used to drive the visual cortex neurons into entrained oscillatory firing. This is compared to a checkerboard flashing aperiodically, with the same average number of flashes per unit time. A magnetoencephalography (MEG) measurement was made to confirm that the periodic paradigm elicited entrainment. We found that for frequencies of 10 and 15 Hz, the periodic stimulus gave a smaller BOLD response than for the aperiodic stimulus. Detailed investigation at 15 Hz showed that the aperiodic stimulus gave a similar BOLD increase regardless of the magnitude of jitter (+/-17 ms compared to +/-33 ms), indicating that flashes need to be precise to at least 17 ms to maintain entrainment. This is also evidence that for aperiodic stimuli, the amplitude of the BOLD response ordinarily reflects the total number of flashes per unit time, irrespective of the precise spacing between them, suggesting that entrainment is the main cause of the BOLD reduction in the periodic condition. The results indicate that, during entrainment, there is a reduction in the neuronal metabolic demand. We suggest that because of the selective frequency band of this effect, it could be connected to synchronised reverberations around an internal feedback loop. PMID- 14741662 TI - OS-EM and FBP reconstructions at low count rates: effect on 3D PET studies of [11C] WAY-100635. AB - 11C-labeled neuroreceptor ligands frequently require long scan durations to quantify ligand-receptor binding. In this paper, we compare the accuracy of two three-dimensional (3D) positron emission tomography (PET) reconstructions: ordered-subset expectation-maximization (OS-EM) versus filtered backprojection (FBP) under low count rate conditions exhibited by 11C neuroreceptor studies. Data were obtained from a dynamic 11C phantom acquisition as well as six dynamic human [11C] WAY-100635 studies, all acquired in 3D mode using the EXACT HR+ PET scanner. Model-based scatter correction of the phantom datum was found to overcorrect in low count rate situations producing a negative bias in FBP reconstruction and a positive bias in OS-EM reconstruction, the OS-EM bias attributed to the non-negativity constraint of sinogram values. In the phantom OS EM and FBP, reconstruction bias occurred at activities less than 25 Bq/cm3. In the human cerebellum, OS-EM deviated from FBP at activities less than 50 Bq/cm3. The total volume of distribution (VT), as determined from the metabolite corrected arterial input function and a two-tissue compartment kinetic model, was more sensitive to the positive bias of OS-EM than the negative bias of FBP at low count rates. To avoid reconstruction bias with 3D PET studies using the HR+, the scan duration should be limited so as to yield a final non-decay-corrected activity concentration of no less than 50 Bq/cm3. In neuroreceptor studies, if such a low count rate cannot be avoided, FBP reconstruction is preferable to OS EM to estimate VT. PMID- 14741663 TI - Application of independent component analysis to magnetic resonance imaging for enhancing the contrast of gray and white matter. AB - An application of independent component analysis (ICA) was attempted to develop a method of processing magnetic resonance (MR) images to extract physiologically independent components representing tissue relaxation times and achieve improved visualization of normal and pathologic structures. Anatomical T1-weighted, T2 weighted and proton density images were obtained from 10 normal subjects, 3 patients with brain tumors and 1 patient with multiple sclerosis. The data sets were analyzed using ICA based on the learning rule of Bell and Sejnowski after prewhitening operations. The three independent components obtained from the three original data sets corresponded to (1) short T1 components representing myelin of white matter and lipids, (2) relatively short T1 components representing gray matter and (3) long T2 components representing free water. The involvement of gray or white matter in brain tumor cases and the demyelination in the case of multiple sclerosis were enhanced and visualized in independent component images. ICA can potentially achieve separation of tissues with different relaxation characteristics and generate new contrast images of gray and white matter. With the proper choice of contrast for the original images, ICA may be useful not only for extracting subtle or hidden changes but also for preprocessing transformation before clustering and segmenting the structure of the human brain. PMID- 14741664 TI - White-matter diffusion anisotropy after chemo-irradiation: a statistical parametric mapping study and histogram analysis. AB - The aim of the study was to evaluate white-matter (WM) diffusion anisotropy in medulloblastoma survivors after cranial irradiation and chemotherapy using voxel based analysis with SPM99 and fractional anisotropy (FA) histogram-derived indices, and to identify quantitative indices for detecting and monitoring children with treatment-induced white-matter injury. Familywise error rate (FWE) that corrects for multiple comparisons was used to locate statistically significant regions of P < 0.05 in voxel-based analysis. Subsequently, the false discovery rate (FDR) controlling procedure (corrected P < 0.05) was used. FA map histogram analysis of histogram-derived indices, mean FA, mean FA peak height, and peak location was performed. Two-sample t test was used in all analyses. Using FWE-corrected P < 0.05, there was a cluster of reduced anisotropy in the periventricular white matter lateral to the left ventricular atrium. When FDR corrected P < 0.05 was used, there were multiple clusters of reduced anisotropy in the periventricular white matter, the corpus callosum, and corona radiata. Simplified voxel-based morphometry (VBM)-like analysis of cerebrospinal fluid (CSF) did not show significant differences between patient and control subjects. 'White-matter FA map' histogram showed significant reduction in mean FA and mean FA peak location and significant increase in mean FA peak height in the patient group compared to control subjects (P = 0.003, P = 0.003, and P = 0.014, respectively). This approach of quantifying FA can be applied to characterize anisotropy in the white matter after cranial irradiation and chemotherapy and can potentially be used to detect and monitor treatment-induced neurotoxicity. PMID- 14741665 TI - A knowledge-driven algorithm for a rapid and automatic extraction of the human cerebral ventricular system from MR neuroimages. AB - A knowledge-driven algorithm for a rapid, robust, accurate, and automatic extraction of the human cerebral ventricular system from MR neuroimages is proposed. Its novelty is in combination of neuroanatomy, radiological properties, and variability of the ventricular system with image processing techniques. The ventricular system is divided into six 3D regions: bodies and inferior horns of the lateral ventricles, third ventricle, and fourth ventricle. Within each ventricular region, a 2D region of interest (ROI) is defined based on anatomy and variability. Each ventricular region is further subdivided into subregions, and conditions detecting and preventing leakage into the extra-ventricular space are specified for each subregion. The algorithm extracts the ventricular system by (1) processing each ROI (to calculate its local statistics, determine local intensity ranges of cerebrospinal fluid and gray and white matters, set a seed point within the ROI, grow region directionally in 3D, check anti-leakage conditions, and correct growing if leakage occurred) and (2) connecting all unconnected regions grown by relaxing growing conditions. The algorithm was validated qualitatively on 68 and quantitatively on 38 MRI normal and pathological cases (30 clinical, 20 MGH Brain Repository, and 18 MNI BrainWeb data sets). It runs successfully for normal and pathological cases provided that the slice thickness is less than 3.0 mm in axial and less than 2.0 mm in coronal directions, and the data do not have a high inter-slice intensity variability. The algorithm also works satisfactorily in the presence of up to 9% noise and up to 40% RF inhomogeneity for the BrainWeb data. The running time is less than 5 s on a Pentium 4, 2.0 GHz PC. The best overlap metric between the results of a radiology expert and the algorithm is 0.9879 and the worst 0.9527; the mean and standard deviation of the overlap metric are 0.9723 and 0.01087, respectively. PMID- 14741666 TI - Towards a standard analysis for functional near-infrared imaging. AB - Functional near-infrared spectroscopy (fNIRS) allows the ability to monitor brain activation by measuring changes in the concentration of oxy- and deoxy hemoglobin. Until now no standardized approach for fNIRS data analysis has been established, although this has to be regarded as a precondition for future application. Hence, we applied the well-established general linear model to optical imaging data. Further, fNIRS data were analyzed in the frequency domain. Two visual tasks were investigated with optical imaging: a checkerboard paradigm supposed to activate the primary and secondary visual cortex, and a paradigm consisting of moving colored stimuli (rotating 'L's) additionally involving the motion area V5. Analysis with the general linear model detected the activation focus in the primary and secondary visual cortex during the first paradigm. For the second paradigm, a second laterally localized activated brain region was found, most likely representing V5. Spatially resolved spectral analysis confirmed the results by showing maxima of power spectral density and coherence in the same respective brain regions. Moreover, it demonstrated a delay of the hemodynamic response in the motion area. In summary, the present study suggests that the general linear model and spatially resolved spectral analysis can be used as standard statistical approaches for optical imaging data, particularly because they are almost independent of the assumed differential path length factors. PMID- 14741667 TI - Comparison of spiral-in/out and spiral-out BOLD fMRI at 1.5 and 3 T. AB - Spiral-in/out functional magnetic resonance imaging (fMRI) methods acquire one image before the echo time (TE) and a second image after TE during each scan. Weighted combination of the two images provides a time series with reduced susceptibility dropout in frontal and medial temporal regions as well as increased signal-to-noise ratio (SNR) in regions of uniform cortex. In this study, task activation with the spiral-in/out method was compared to that with conventional spiral-out acquisitions at two field strengths (1.5 and 3.0 T) using episodic memory encoding, verbal working memory, and affective processing tasks in eight human volunteers. With the conventional spiral-out sequence, greater signal dropout is observed in lateral and medial prefrontal, amygdalar, and medial temporal regions at 3 T relative to 1.5 T, whereas such dropout at 3 T is reduced or mitigated with the spiral-in/out method. Similarly, activation volumes for frontal, amygdalar, and medial temporal regions are reduced for spiral-out acquisitions relative to spiral-in/out, and this difference is more apparent at 3 T than at 1.5 T. In addition, significant regionally specific increases in Z scores are obtained with the spiral-in/out sequence relative to spiral-out acquisitions at both field strengths. It is concluded the spiral-in/out sequence may provide significant advantages over conventional spiral methods, especially at 3 T. PMID- 14741668 TI - Probability effects on the neural correlates of retrieval success: an fMRI study. AB - Event-related fMRI was employed to investigate the influence of the relative probability of old and new test items on the neural correlates of recognition memory. Twelve subjects undertook three study-test cycles, each consisting of an identical study phase in which a series of words was encoded in an incidental task, followed by a test phase in which yes/no recognition judgments were made to a mixture of studied (old) and unstudied (new) words. The ratio of old to new words differed in each test phase, and was either 25:75, 50:50, or 75:25. In lateral inferior and medial parietal cortex, and the posterior cingulate, greater activity was elicited by correctly classified old than new items independently of old:new ratio. By contrast, in other regions, including anterior, dorsolateral, and ventrolateral prefrontal cortex, differences in the activity elicited by old and new items varied according to old:new ratio, demonstrating in some cases a complete crossover interaction. The results suggest that differential activity elicited by old and new test items is likely to support successful recognition in only a subset of the regions identified in previous studies as exhibiting such differences. In other regions, most notably prefrontal cortex, differences in the activity elicited by old and new items appear to reflect processes that are contingent upon, rather than in support of, successful recognition. PMID- 14741669 TI - Mapping transplanted stem cell migration after a stroke: a serial, in vivo magnetic resonance imaging study. AB - Preferential migration of stem cells toward the site of a lesion is a highly desirable property of stem cells that allows flexibility in the site of graft implantation in the damaged brain. In rats with unilateral stroke damage, neural stem cells transplanted into the contralateral hemisphere migrate across to the lesioned hemisphere and populate the area around the ischaemic infarct. To date, the migration of neural stem cells in the damaged brain has been mainly inferred from snapshot histological images. In this study, we demonstrate that by pre labelling neural stem cells with the bimodal contrast agent Gadolinium-RhodamIne Dextran [GRID, detectable by both magnetic resonance imaging (MRI) and fluorescent microscopy], the transhemispheric migration of transplanted neural stem cells contralateral to a stroke lesion can be followed in vivo by serial MRI and corroborated by subsequent histological analyses. Our results indicate that neural stem cells migrated from the injection tract mainly along the corpus callosum within 7 days of transplantation and extensively re-populated the peri lesion area by 14 days following implantation. In contrast, neural stem cells transplanted into sham controls did not show any substantial migration outside of the injection tract, suggesting that the transcallosal migration observed in the stroke-lesioned animals is due to neural stem cells being attracted by the lesion site. In vivo tracking of the migration of neural stem cells responding to damage will greatly enhance our understanding of optimal transplantation strategies as well as how neural stem cells promote functional and anatomical recovery in neurological disorders. PMID- 14741670 TI - Cerebral correlates of alerting, orienting and reorienting of visuospatial attention: an event-related fMRI study. AB - The identification of brain systems contributing to different aspects of visuospatial attention is of both clinical and theoretical interest. Cued target detection tasks provide a simple means to dissociate attentional subcomponents, such as alerting, orienting or reorienting of attention. Event-related functional magnetic resonance imaging (fMRI) was used to study neural correlates of these distinct attentional processes. Volunteers were scanned while performing a centrally cued target detection task. Four different types of trials (no cue, neutral cue, valid cue and invalid cue trials) with targets appearing either in the right or left hemifield were randomly intermixed. Behaviourally, the data provided evidence for alerting, spatial orienting and reorienting of attention. Neurally, the alerting effect was seen in bilaterally increased extrastriatal blood oxygenation level-dependent (BOLD) activity in neutral as compared to no cue trials. Neural correlates of spatial orienting were seen in anterior cingulate cortex, which was more active during valid as compared to neutral cue trials. Neural correlates of reorienting of attention, that is, higher BOLD activity to invalid as compared to validly cued trials were evident in several brain regions including left and right intraparietal sulcus, right temporo parietal junction and middle frontal gyrus bilaterally. The data suggest that frontal and parietal regions are specifically involved in reorienting rather than orienting attention to a spatial position. Alerting effects were seen in extrastriate regions which suggest that increased phasic alertness results in a top-down modulation of neural activity in visual processing areas. PMID- 14741671 TI - A functional MRI study of visual oddball: evidence for frontoparietal dysfunction in subjects at risk for alcoholism. AB - Attending to rare stimuli interspersed among repetitive frequent stimuli produces a positive scalp potential at 300 to 600 ms after the target stimulus onset; this potential is known as the P300 wave. Although there is clear evidence of low visual P300 in subjects at high risk (HR) for developing alcoholism, the functional neuroanatomical correlates have not been studied. Functional and high resolution anatomical magnetic resonance images were collected during the performance of a visual oddball task, from six control (low risk-LR) subjects with high P300s and eight HR subjects with low P300s. All the HR subjects were offspring of male alcoholics. The data were analyzed using a randomization-based statistical method that accounts for multiple comparisons, requires no assumptions about the noise structure of the data, and does not require spatial or temporal smoothing. Target counts showed that all subjects performed the task comparably. Analysis of the functional magnetic resonance imaging (fMRI) data revealed two areas with significantly lower activation in the HR group when compared to the LR group: the bilateral inferior parietal lobule (BA 40), and the bilateral inferior frontal gyrus (BA 44). Inferior parietal lobule showed significantly lower activation in the HR group in contrast to the LR group, and inferior frontal gyrus was not activated in the HR group but was only activated in the LR group. This finding indicates that a dysfunctional frontoparietal circuit may underlie the low P300 responses seen in HR subjects. This perhaps implies a deficiency in the rehearsal component of the working memory system. PMID- 14741672 TI - Modality effects in verbal working memory: differential prefrontal and parietal responses to auditory and visual stimuli. AB - The neural bases of verbal (nonspatial) working memory (VWM) have been primarily examined using visual stimuli. Few studies have investigated the neural bases of VWM using auditory stimuli, and fewer have explored modality differences in VWM. In this study, we used functional magnetic resonance imaging (fMRI) to examine similarities and differences between visual VWM (vis-VWM) and auditory VWM (aud VWM) utilizing identical stimuli and a within-subjects design. Performance levels were similar in the two modalities and there was extensive overlap of activation bilaterally in the dorsolateral and ventrolateral prefrontal cortex (DLPFC and VLPFC), intraparietal sulcus, supramarginal gyrus and the basal ganglia. However, a direct statistical comparison revealed significant modality differences: the left posterior parietal cortex, primarily along the intraparietal sulcus, showed greater responses during vis-VWM whereas the left dorsolateral prefrontal cortex showed greater responses during aud-VWM. No such differences were observed in the right hemisphere. Other modality differences in VWM were also observed, but they were associated with relative decreases in activation. In particular, we detected bilateral suppression of the superior and middle temporal (auditory) cortex during vis-VWM, and of the occipital (visual) cortex during aud-VWM, thus suggesting that cross-modal inhibitory processes may help to provide preferential access to high-order heteromodal association areas. Taken together, our findings suggest that although similar prefrontal and parietal regions are involved in aud VWM and vis-VWM, there are important modality differences in the way neural signals are generated, processed and routed during VWM. PMID- 14741673 TI - Functional association of the amygdala and ventral prefrontal cortex during cognitive evaluation of facial expressions primed by masked angry faces: an event related fMRI study. AB - The present study examined the functional association of the amygdala and right ventral prefrontal cortex (PFC) during cognitive evaluation of facial expressions. A situation was created where emotional valence of the stimuli was unconsciously manipulated by using subliminal affective priming. Twelve healthy volunteers were asked to evaluate the facial expressions of a target face (500-ms duration) such as "anger", "neutral", or "happy". All target faces expressed relatively weak anger. Just before the presentation of the target face, a prime of three conditions of 35-ms duration, angry face, neutral face, and white blank was presented. The subjects could not consciously identify the primes in this procedure. Activity in the right amygdala was greater with subliminal presentation of the angry prime compared with subliminal presentation of a neutral face or white-blank stimuli. Most importantly, the degree of activation of the right amygdala was negatively correlated with that of the right ventral PFC only with the anger prime. Furthermore, activation of the amygdala was positively correlated with rate of judgment when the subjects recognized anger in the target faces. These results are discussed in terms of the functional association between the right PFC and the amygdala and its influence on cognitive processing. PMID- 14741674 TI - Estrogen- and tamoxifen-associated effects on brain structure and function. AB - We evaluated the effects of estrogen and tamoxifen, a selective estrogen receptor modulator, on positron emission tomography (PET) measures of brain glucose metabolism and magnetic resonance imaging (MRI) measures of hippocampal atrophy. Three groups of postmenopausal women were studied, women taking estrogen (ERT+), women with breast cancer taking tamoxifen (TAM), and women not taking estrogen or tamoxifen (ERT-). All subjects received a PET scan, an MRI scan, and cognitive testing. The TAM group showed widespread areas of hypometabolism in the inferior and dorsal lateral frontal lobes relative to the other two groups. The ERT- group showed lower metabolism in the inferior frontal cortex and temporal cortex with respect to the ERT+ group. The TAM group also showed significantly lower semantic memory scores than the other two groups. Finally, the TAM group had smaller right hippocampal volumes than the ERT+ group, an effect that was of borderline significance. Both right and left hippocampal volumes were significantly smaller than the ERT+ group when a single outlier was removed. The ERT- group had hippocampal volumes that were intermediate to the other two groups. These findings provide physiological and anatomical evidence for neuroprotective effects of estrogen. PMID- 14741675 TI - Noninvasive measurement of neuronal activity with near-infrared optical imaging. AB - Diffuse optical imaging (DOI) alone offers the possibility of simultaneously and noninvasively measuring neuronal and vascular signals in the brain with temporal resolution of up to 1 ms. However, while optical measurement of hemodynamic signals is well established, optical measurement of neuronal activation (the so called fast signal) is just emerging and requires further optimization and validation. In this work, we present preliminary studies in which we measured the fast signal in 10 healthy volunteers during finger-tapping, tactile stimulation, and electrical median nerve stimulation. We used an instrument (CW4) with 8 source (690 and 830 nm) and 16 detector positions-more optodes than the instruments in previously reported studies. This allowed us to record the ipsilateral and contralateral sensorimotor cortex simultaneously, while at the same time measuring the evoked hemodynamic response. We used an acquisition time of 25 ms per image; after averaging approximately 1000 events, the signal-to noise ratio was approximately 10(4). Since the expected relative intensity changes due to the fast signal (approximately 10(-3)) are smaller than the relative intensity changes due to physiological effects (approximately 10(-1)), we enhanced the suppression of competing signals such as the heartbeat-associated intensity changes, and established five criteria with which to assess the robustness of the fast signal. We detected the fast signal in 43% of the measurements during finger-tapping, 60% of those during tactile stimulation, and 23% of those during electrical median nerve stimulation. The relative changes in intensity associated with the fast signal were approximately 0.07% and the latency of the signal was approximately 100 ms. PMID- 14741676 TI - Efficiency, power, and entropy in event-related FMRI with multiple trial types. Part I: theory. AB - Experimental designs for functional magnetic resonance imaging (fMRI) experiments can be characterized by their estimation efficiency, which is a measure of the variance in the estimate of the hemodynamic response function (HRF), and their detection power, which is a measure of the variance in the estimate of the amplitude of functional activity. Previous studies have shown that there exists a fundamental trade-off between efficiency and power for experiments with a single trial type of interest. This paper extends the prior work by presenting a theoretical model for the relation between detection power and estimation efficiency in experiments with multiple trial types. It is shown that the trade off between efficiency and power present in multiple-trial-type experiments is identical in form to that observed for single-trial-type experiments. Departures from the predicted trade-off due to the inclusion of basis function expansions and the assumption of correlated noise are examined. Finally, conditional entropy is introduced as measure for the randomness of a design, and an empirical relation between entropy and estimation efficiency is presented. PMID- 14741677 TI - Efficiency, power, and entropy in event-related fMRI with multiple trial types. Part II: design of experiments. AB - The performance of an experimental design for functional magnetic resonance imaging (fMRI) can be characterized by its estimation efficiency, which is the ability to make an estimate of the hemodynamic response, its detection power, which is the ability to detect an activation, and its conditional entropy, which is a measure of the randomness of the design. In Liu and Frank [Neuroimage 21 (2004) 387-400], it is shown that there is a fundamental theoretical trade-off between estimation efficiency and detection power for experiments with multiple trial types and that there is an empirical relation between estimation efficiency and conditional entropy. This paper provides an intuitive interpretation of the theoretical results and examines the practical implications of these results for the optimal design of fMRI experiments with multiple trial types. The properties of block designs, permuted block designs, m-sequence designs, clustered m sequence designs, and mixed designs are explored. It is shown that these designs nearly achieve the theoretically predicted performance and can be used in practice to obtain advantageous trade-offs among efficiency, power, and entropy. PMID- 14741678 TI - Emotional valence of words modulates the subliminal repetition priming effect in the left fusiform gyrus: an event-related fMRI study. AB - The present study investigated if the emotional valence of words modulates the subliminal repetition priming effect in the brain, in particular, the occipitotemporal visual cortex, by adopting a rapid presentation event-related fMRI design. A masked repetition priming paradigm was adopted, in which, before the presentation of the target (either positive or negative or neutral in meaning), a masked prime word that was either a repetition or an unrelated word of the target was presented. The subject made a perceptual judgment on the target. The results revealed that the left mid-fusiform gyrus was sensitive to the emotional manipulation of the repetition priming effect and that the priming effect in the region was greater in the positive than in the negative word condition. The priming effect in the fusiform gyrus in neutral words was not significant, which might be a result of suppression caused by the emotional context. No effect of valence or repetition was found in the amygdala. PMID- 14741679 TI - Interindividual variability in the hemispheric organization for speech. AB - A PET activation study was designed to investigate hemispheric specialization during speech comprehension and production in right- and left-handed subjects. Normalized regional cerebral blood flow (NrCBF) was repeatedly monitored while subjects either listened to factual stories (Story) or covertly generated verbs semantically related to heard nouns (Gener), using silent resting (Rest) as a common control condition. NrCBF variations in each task, as compared to Rest, as well as functional asymmetry indices (FAI = right minus left NrCBF variations), were computed in anatomical regions of interest (AROIs) defined on the single subject MNI template. FAIs were predominantly leftward in all regions during both tasks, although larger FAIs were observed during Gener. Subjects were declared "typical" for language hemispheric specialization based on the presence of significant leftward asymmetries (FAI < 0) in the pars triangularis and opercularis of the inferior frontal gyrus during Gener, and in the middle and inferior temporal AROIs during Story. Six subjects (including five LH) showed an atypical language representation. Among them, one presented a right hemisphere specialization during both tasks, another a shift in hemispheric specialization from production to comprehension (left during Gener, right during Story). The group of 14 typical subjects showed significant positive correlation between homologous left and right AROIs NrCBF variations in temporal areas during Story, and in temporal and inferior frontal areas during Gener, almost all regions presenting a leftward FAI. Such correlations were also present in deactivated areas with strong leftward asymmetry (supramarginalis gyrus, inferior parietal region). These results suggest that entry into a language task translates into a hemispheric reconfiguration of lateral cortical areas with global NrCBF increase in the dominant hemisphere and decrease in the minor hemisphere. This can be considered as the setting up of a "language mode", under the control of a mechanism that operates at a perisylvian level. On top of this global organization, regional variations carry on the performance of the cognitive operations specific to the language task to be performed. Hemispheric relationships could be different in atypical subjects, with either between task hemispheric regulation differences or differences in regional specialization. PMID- 14741680 TI - Learned regulation of spatially localized brain activation using real-time fMRI. AB - It is not currently known whether subjects can learn to voluntarily control activation in localized regions of their own brain using neuroimaging. Here, we show that subjects were able to learn enhanced voluntary control over task specific activation in a chosen target region, the somatomotor cortex. During an imagined manual action task, subjects were provided with continuous direction regarding their cognitive processes. Subjects received feedback information about their current level of activation in a target region of interest (ROI) derived using real-time functional magnetic resonance imaging (rtfMRI), and they received automatically-adjusted instructions for the level of activation to achieve. Information was provided both as continously upated graphs and using a simple virtual reality interface that provided an image analog of the level of activation. Through training, subjects achieved an enhancement in their control over brain activation that was anatomically specific to the target ROI, the somatomotor cortex. The enhancement took place when rtfMRI-based training was provided, but not in a control group that received similar training without rtfMRI information, showing that the effect was not due to conventional, practice based neural plasticity alone. Following training, using cognitive processes alone subjects could volitionally induce fMRI activation in the somatomotor cortex that was comparable in magnitude to the activation observed during actual movement. The trained subjects increased fMRI activation without muscle tensing, and were able to continue to control brain activation even when real-time fMRI information was no longer provided. These results show that rtfMRI information can be used to direct cognitive processes, and that subjects are able to learn volitionally regulate activation in an anatomically-targeted brain region, surpassing the task-driven activation present before training. PMID- 14741681 TI - Epilepsy surgery, resection volume and MSI localization in lesional frontal lobe epilepsy. AB - To verify whether interictal noninvasive information detected by magnetoencephalography (MEG) recordings can contribute to localize focal epileptic activity relevant for seizure generation in lesional frontal lobe epilepsy, magnetic source imaging (MSI) localizations of epileptic discharges were compared to the extent of neurosurgical resection and postoperative outcome. Preoperative MEG spike localizations were displayed in postoperative magnetic resonance imaging (MRI) scans to check whether dipole sites were located within the resection cavity. Moreover, MEG localizations were compared with results of prolonged video-EEG monitoring and, in three cases, with invasive EEG recordings. Our results in five cases with lesional frontal lobe epilepsy showed that good surgical outcome could be achieved in those patients where the majority of MEG spike localizations were located within the resected brain volume. PMID- 14741682 TI - Precentral gyrus discrepancy in electronic versions of the Talairach atlas. AB - Electronic versions of the atlas of Talairach and Tournoux, including the Talairach Daemon and the official versions published by Thieme, contain a discrepant region of the precentral gyrus on axial slice +35 mm that extends far forward into the frontal lobe. This area is anatomically incorrect and internally inconsistent within the digital atlas software applications using their multiplanar cross-referencing tools. By cross-referencing the axial, sagittal, and coronal plates from the original printed atlas, we demonstrate that the discrepant area should be labeled middle frontal gyrus. The mislabeled portion encompasses a 3 x 1.5-cm region in the axial plane and has significant implications for sensorimotor studies that rely on the digital atlases for anatomic labeling. PMID- 14741683 TI - Encoding activity in anterior medial temporal lobe supports subsequent associative recognition. AB - The ability to bind information together, such as linking a name with a face or a car with a parking space, is a vital process in human episodic memory. To identify the neural bases for this binding process, we measured brain activity during a verbal associative encoding task using event-related functional MRI (fMRI), followed by an associative recognition test for the studied word pairs. Analysis of the encoding data sorted by the associative recognition accuracy allowed us to isolate regions involved in successfully creating associations. We found that encoding activity in bilateral anterior medial temporal lobe (MTL) regions was greater for successfully bound pairs, that is, those later recognized as intact, than for all other pairs. These findings provide evidence that the anterior medial temporal lobes support the successful binding of information in memory. PMID- 14741684 TI - Combination of event-related fMRI and diffusion tensor imaging in an infant with perinatal stroke. AB - Focal ischemic brain injury, or stroke, is an important cause of later handicap in children. Early assessment of structure-function relationships after such injury will provide insight into clinico-anatomic correlation and potentially guide early intervention strategies. We used combined functional MRI (fMRI) with diffusion tensor imaging (DTI) in a 3-month-old infant to explore the structure function relationship after unilateral perinatal stroke that involved the visual pathways. With visual stimuli, fMRI showed a negative BOLD activation in the visual cortex of the intact right hemisphere, principally in the anterior part, and no activation in the injured hemisphere. The functional activation in the intact hemisphere correlated clearly with the fiber tract of the optic radiation visualized with DTI. DTI confirmed the absence of the optic radiation in the damaged left hemisphere. In addition, event-related fMRI (ER-fMRI) experiments were performed to define the characteristics of the BOLD response. The shape is that of an inverted gamma function (similar to a negative mirror image of the known positive adult BOLD response). The maximum decrease was reached at 5-7 s with signal changes of -1.7 +/- 0.4%.Thus, this report describes for the first time the combined use of DTI and event-related fMRI in an infant and provides insight into the localization of the fMRI visual response in the young infant and the characteristics of the BOLD response. PMID- 14741685 TI - Resting functional MRI with temporal clustering analysis for localization of epileptic activity without EEG. AB - We report on the methods and initial findings of a novel noninvasive technique, resting functional magnetic resonance imaging (fMRI) with temporal clustering analysis (TCA), for localizing interictal epileptic activity. Nine subjects were studied including six temporal lobe epilepsy (TLE) patients with confirmed localization indicated by successful seizure control after resection. The remaining three subjects had standard presurgical evaluations with inconsistent results or suspected extratemporal lobe foci. Peaks of activity, presumably epileptic, were detected in all nine subjects, using the resting functional MRI with temporal clustering analysis. In all six patients who underwent resective surgery, the fMRI with temporal clustering analysis accurately determined the epileptogenic hippocampal hemisphere (P = 0.005). In the three subjects without confirmed localization, the technique determined regions of activity consistent with those determined by the presurgical assessments. Though more studies are required to validate this technique, the results demonstrate the potential of the resting fMRI with temporal clustering technique to detect and localize epileptic activity without the need for simultaneous electroencephalography (EEG). The greatest potential benefit of this technique will be in the evaluation of patients with suspected extratemporal lobe epilepsy and patients whose standard assessments are discordant. PMID- 14741686 TI - Role of phenytoin in wound healing: microarray analysis of early transcriptional responses in human dermal fibroblasts. AB - Wound healing is a complex process involving a number of related genes and receptors. Using cDNA microarrays, we explored the global gene expression profile of phenytoin (20microg/ml) induced changes to human dermal fibroblasts. Microarray data analysis revealed approximately 1500 genes were differentially expressed by 2.5-fold. At 3, 6, 12, and 24h, the transcripts of the major growth factors involved in wound healing and their receptors were increased. This was further confirmed by RT-PCR. Genes encoding other proteins with roles in signal transduction (NFkappaB), extracellular matrix (MMP1) including type I collagen, fibronectin, and laminin were strongly induced at 6h and onwards. Genes involved in cell cycle regulation (CCND1 and CDKN1A) were down-regulated consistent with our finding that phenytoin per se did not have cell proliferation activity. Notably, phenytoin accelerates the autocrine and paracrine activity of growth factors by up-regulating the related receptors. PMID- 14741687 TI - Removal of clustered positive charge from dihydropyridine receptor II-III loop peptide augments activation of ryanodine receptors. AB - Peptides based on the skeletal muscle DHPR II-III loop have been shown to regulate ryanodine receptor channel activity. The N-terminal region of this cytoplasmic loop is predicted to adopt an alpha-helical conformation. We have selected a peptide sequence of 26 residues (Ala(667)-Asp(692)) as the minimum sequence to emulate the helical propensity of the corresponding protein sequence. The interaction of this control peptide with skeletal and cardiac RyR channels in planar lipid bilayers was then assessed and was found to lack isoform specificity. At low concentrations peptide A(667)-D(692) increased RyR open probability, whilst at higher concentrations open probability was reduced. By replacing a region of clustered positive charge with a neutral sequence with the same predisposition to helicity, the inhibitory effect was ablated and activation was enhanced. This novel finding demonstrates that activation does not derive from the presence of positively charged residues adjacent in the primary structure and, although it may be mediated by the alignment of basic residues down one face of an amphipathic helix, not all of these residues are essential. PMID- 14741688 TI - Selective in vitro antioxidant properties of bisphosphonates. AB - The aim of this study was to investigate the in vitro antioxidant profile of different bisphosphonates. Bisphosphonates were tested for their xanthine oxidase and microsomal lipid peroxidation inhibiting capacity. Furthermore, the effect of these different compounds on DPPH, a stable radical, was investigated. Clodronate, risedronate, and pyrophosphate were further tested for their hydroxyl radical scavenging activity. None of the tested compounds showed xanthine oxidase inhibiting activity or DPPH scavenging activity. All the tested bisphosphonates exhibited inhibiting capacities on the microsomal lipid peroxidation. The hydroxyl radical scavenging activity was dependent on the order of adding the different reagents and was highest for risedronate. Bisphosphonates possess an inhibiting activity on the microsomal lipid peroxidation and the Fenton reaction. In these reactions iron plays an important role suggesting that the selective in vitro antioxidant properties of the bisphosphonates are due to their iron chelating characteristics. PMID- 14741689 TI - A single WAP domain-containing protein from Litopenaeus vannamei hemocytes. AB - A cDNA clone coding for a single WAP domain (SWD) protein was isolated from a hemocyte cDNA library of Litopenaeus vannamei. The full-length cDNA sequence is 0.4kb long and encodes a 93-amino acid protein. Using this sequence as a probe a similar clone coding for a 92-amino acids protein was found in a cDNA library from Penaeus monodon hemocytes. The mRNA size was confirmed by Northern blot as well as that gene is expressed in hemocytes, but not in hepatopancreas. mRNA levels of the shrimp SWD protein were modified after injection of Vibrio alginolyticus, indicating the probable role of this protein in the immune response. Although amino acid sequence seems to be similar to those of other WAP domain-containing proteins, shrimp SWD protein does not have any other functional domain, similar to a mouse single WAP motif (SWAM) protein reported in mouse; however, the phylogenetic analysis shows that shrimp SWD is more related to other WAP proteins than to mouse SWAM. PMID- 14741690 TI - Protein kinase PKN1 associates with TRAF2 and is involved in TRAF2-NF-kappaB signaling pathway. AB - PKN1 is a fatty acid and Rho-activated serine/threonine protein kinase whose catalytic domain is highly homologous to protein kinase C (PKC) family. In yeast two-hybrid screening for PKN1 binding proteins, we identified tumor necrosis factor alpha (TNFalpha) receptor-associated factor 2 (TRAF2). TRAF2 is one of the major mediators of TNF receptor superfamily transducing TNF signal to various functional targets, including activation of NF-kappaB, JNK, and apoptosis. FLAG tagged PKN1 was co-immunoprecipitated with endogenous TRAF2 from HEK293 cell lysate, and in vitro binding assay using the deletion mutants of TRAF2 showed that PKN1 directly binds to the TRAF domain of TRAF2. PKN1 has the TRAF2-binding consensus sequences PXQX (S/T) at amino acid residues 580-584 (PIQES), and P580AQ582A mutant was not co-immunoprecipitated with TRAF2. Furthermore, the reduced expression of PKN1 by RNA interference (RNAi) down-regulated TRAF2 induced NF-kappaB activation in HEK293T cells. These results suggest that PKN1 is involved in TRAF2-NF-kappaB signaling pathway. PMID- 14741691 TI - Ca2+/calmodulin-dependent protein phosphatase calcineurin mediates the expression of iNOS through IKK and NF-kappaB activity in LPS-stimulated mouse peritoneal macrophages and RAW 264.7 cells. AB - Ca(2+) and Ca(2+)/calmodulin-dependent protein phosphatase calcineurin (CN) have been known to play crucial roles in immune response and inflammation. Using mouse peritoneal macrophages and RAW 264.7 macrophage cells, we demonstrated that LPS mobilized intracellular free Ca(2+) and induced CN phosphatase activity. iNOS expression and NO secretion in response to LPS were suppressed by Ca(2+) antagonists (TMB-8, BAPTA/AM, and nifedipine) and CN inhibitor (cyclosporin A). Transient expression of constitutively active CN in mouse peritoneal macrophages and RAW 264.7 macrophages strongly activated NF-kappaB, a key mediator of iNOS expression. We also found that CN mediates NF-kappaB activation via IkappaB-alpha hyperphosphorylation and degradation. Overexpression of dominant negative mutant of IKKalpha and -beta demonstrates that only IKKbeta is the target for CN. These results indicate that CN is required for full iNOS expression and the effective activation of NF-kappaB in RAW 264.7 and peritoneal macrophages. PMID- 14741692 TI - Pravastatin up-regulates transforming growth factor-beta1 in THP-1 human macrophages: effect on scavenger receptor class A expression. AB - Statins have been shown to interact with several monocyte/macrophage functions. We tested the effect of pravastatin on transforming growth factor-beta1 (TGF beta1) production and its possible involvement in scavenger receptors class A (SRA) expression in human THP-1 cells. TGF-beta1s biological activity in THP-1 cell conditioned medium, evaluated by luciferase activity of transfected cell with a TGF-beta responsive promoter, was increased in a dose-dependent manner after incubation with pravastatin (1-20 microM). Pravastatin (1-20 microM) induced a dose-dependent increase in TGF-beta1 mRNA expression and protein production in THP-1 cells. PMA-induced SRA gene and protein expression was suppressed by pravastatin with a mean 3-fold decrease at 10 microM. This last effect was reversed by a mouse monoclonal anti-TGF-beta1 neutralizing antibody. PD98059, a specific inhibitor of MAP kinase cascade, completely reversed pravastatin-induced SRA down-regulation. p44 and p42 isoforms showed a dose dependent phosphorylation after treatment with pravastatin (1-20 microM) which was inhibited by a mouse monoclonal anti-TGF-beta1 antibody. Our results demonstrate that pravastatin significantly up-regulates TGF-beta1 expression which may be in involved in down-regulation of SRA expression in THP-1 cell cultures. A new pathway for pravastatin effects in atherogenesis can be suggested. PMID- 14741693 TI - Green fluorescent protein causes mitochondria to aggregate in the presence of the Bcl-2 family proteins. AB - Green fluorescent protein (GFP) has been widely used in a variety of experiments in cell biology. When cells were co-transfected with the GFP gene and the bcl-2 family genes bcl-2, bcl-x(L), and bax, mitochondria appeared to aggregate at the periphery of the nucleus specifically where GFP was expressed. Little aggregation was seen in the presence of other members of the GFP family, EGFP (enhanced GFP), ECFP (enhanced cyan variant), and EYFP (enhanced yellow-green variant). GFP but not EGFP seemed to promote cell death induced by pro-apoptotic Bax. Thus, GFP specifically promotes the aggregation of mitochondria when co-expressed with a member of the Bcl-2 family in association with apoptosis. PMID- 14741694 TI - Quantification of cell hybridoma yields with confocal microscopy and flow cytometry. AB - The fusion of antigen presenting and cancer cells leads to the formation of hybrid cells, which are considered a potential vaccine for treating cancer. The quality assessment of hybrid cell vaccines is crucial for the introduction of this new treatment. Flow cytometry was the method used recently, since it is faster in comparison to classical microscopy. Here we describe a rapid confocal microscopy based approach to quantify hybrid cell yields. The extent of fusion rate was determined by confocal microscopy by counting dual fluorescent cells and by measuring the area of co-localized pixels. Results of both methods showed high degree of correlation. The same samples were also analyzed by flow cytometry. Fusion rates determined with both techniques showed significant correlation. In conclusion, using confocal microscopy we developed a sensitive and a rapid method to assess the yield of hybridomas in a large number of electrofused cells. PMID- 14741695 TI - Drought acclimation reduces O2*- accumulation and lipid peroxidation in wheat seedlings. AB - Abiotic stresses cause ROS accumulation, which is detrimental to plant growth. It is well known that acclimation of plants under mild or sub-lethal stress condition leads to development of resistance in plants to severe or lethal stress condition. The generation of ROS and subsequent oxidative damage during drought stress is well documented in the crop plants. However, the effect of drought acclimation treatment on ROS accumulation and lipid peroxidation has not been examined so far. In this study, the effect of water stress acclimation treatment on superoxide radical (O(2)(-z.rad;)) accumulation and membrane lipid peroxidation was studied in leaves and roots of wheat (Triticum aestivum) cv. C306. EPR quantification of superoxide radicals revealed that drought acclimation treatment led to 2-fold increase in superoxide radical accumulation in leaf and roots with no apparent membrane damage. However under subsequent severe water stress condition, the leaf and roots of non-acclimated plants accumulated significantly higher amount of superoxide radicals and showed higher membrane damage than that of acclimated plants. Thus, acclimation-induced restriction of superoxide radical accumulation is one of the cellular processes that confers enhanced water stress tolerance to the acclimated wheat seedlings. PMID- 14741696 TI - Mammalian D-aspartyl endopeptidase: a scavenger for noxious racemized proteins in aging. AB - The accumulation of D-isomers of aspartic acid (D-Asp) in proteins during aging has been implicated in the pathogenesis of Alzheimer's disease, cataracts, and arteriosclerosis. Here, we identified a specific lactacystin-sensitive endopeptidase that cleaves the D-Asp-containing protein and named it D-aspartyl endopeptidase (DAEP). DAEP has a multi-complex structure (MW: 600kDa) and is localized in the inner mitochondrial membrane of mouse and rabbit, but DAEP activity was not detected in Escherichia coli, Saccharomyces cerevisiae, and Caenorhabditis elegans. A specific inhibitor for DAEP was newly synthesized, and inhibited DAEP activity (IC(50), 3microM), a factor of 10 greater than lactacystin on DAEP. On the other hand, the inhibitor did not inhibit either the 20S or 26S proteasome. PMID- 14741698 TI - Secreted brevican mRNA is expressed in the adult rat pituitary. AB - Brevican is the most abundant chondroitin sulfate proteoglycan in the extracellular matrix (ECM) of the adult rat brain. It has been found only in the central nervous system (CNS). In this study, we found that secreted brevican transcript was detectable in the pituitary of both male and female adult rats by reverse transcriptase-polymerase chain reaction (RT-PCR) amplification. In posterior lobe of pituitary, pituicytes were heavily labelled. In anterior and intermediate lobes of pituitary, signals for brevican transcripts were observed in cells of various sizes. These data demonstrated that secreted brevican mRNA is expressed in the adult rat pituitary and brevican might not be a CNS-specific ECM. PMID- 14741697 TI - Replacement of 198MQMDII203 of mouse IRF-1 by 197IPVEVV202 of human IRF-1 abrogates induction of IFN-beta, iNOS, and COX-2 gene expression by IRF-1. AB - Interferon regulatory factor-1 (IRF-1) is a transcription factor exhibiting functional diversity because of its ability to activate transcription from promoters of several IRF-1-dependent genes. It is a modular protein, where the overall structure is not essential for function of its individual domains. A comparison of the mouse and human IRF-1 amino acid sequences enabled us to identify a stretch of six amino acids (198-203) within the transactivation domain of mouse IRF-1, 198MQMDII(203) to be different from that of the human IRF-1, 197IPVEVV(202). This indicated a possible functional significance of the six amino acid stretches in the two IRF-1 molecules. The murine IRF-1 sequence at 198 203 (MQMDII) was replaced by IPVEVV. Recombinant wild type mouse IRF-1 with 198MQMDII(203) and its mutant form with 198IPVEVV(203), expressed as GST-IRF-1 fusion proteins, showed similar DNA-binding activity. However, ectopic expression of the wild type and mutant IRF-1 in the human embryonic kidney (HEK-293) cells showed the effect of replacement of this region on expression of a few chromosomal genes that are transcriptionally activated by IRF-1 viz. IFN-beta, iNOS, and COX-2 genes. In our study, expression of wild type IRF-1 activated these genes as judged by RT-PCR but the mutant IRF-1 did not show this effect. Thus, the MQMDII (198-203 a.a.) region of mouse IRF-1 has a functional context in relation to expression of IRF-1-inducible genes. PMID- 14741699 TI - Beta- and alpha-adrenergic cross-signaling for L-type Ca current is impaired in transgenic mice with constitutive activation of epsilonPKC. AB - It is well established that beta-adrenoceptor stimulation activates PKA and alpha(1)-adrenoceptor stimulation activates PKC. In normal ventricular myocytes, acute activation of alpha(1)-adrenoceptors inhibits beta-adrenoceptor stimulated L-type Ca current (I(Ca-L)) and direct activation of epsilonPKC leads to I(Ca-L) inhibition. Because increased PKC activity has been observed chronically in in vivo setting such as failing human heart, we hypothesized that chronic in vivo activation of epsilonPKC alters I(Ca-L) and its response to adrenergic stimulation. Therefore, we investigated the interaction between beta- and alpha(1)-adrenoceptors vis-a-vis I(Ca-L) in myocytes from transgenic mice (TG) with cardiac specific constitutive activation of epsilonPKC (epsilonPKC agonist). Whole-cell I(Ca-L) was recorded from epsilonPKC agonist TG mice and age-matched non-TG (NTG) littermates under: (1) basal condition, (2) beta-adrenoceptor agonist, isoproterenol (ISO), and (3) ISO plus alpha(1)-adrenoceptor agonist, methoxamine. The present results are the first to demonstrate that chronic in vivo activation of epsilonPKC leads to reduced basal I(Ca-L) density. beta adrenoceptor activation of I(Ca-L) is blunted in epsilonPKC agonist TG mice. alpha-adrenoceptor cross-talk with beta-adrenoceptor signaling pathways vis-a-vis L-type Ca channels is impaired in epsilonPKC agonist TG mice. The diminished response to ISO and methoxamine suggests a protective feedback regulatory mechanism in epsilonPKC agonist TG mice and could be vital in the settings of excessive release of catecholamines during heart failure. PMID- 14741700 TI - Molecular mechanism of enzyme inhibition: prediction of the three-dimensional structure of the dimeric trypsin inhibitor from Leucaena leucocephala by homology modelling. AB - Serine proteinase inhibitors are widely distributed in nature and inhibit the activity of enzymes like trypsin and chymotrypsin. These proteins interfere with the physiological processes such as germination, maturation and form the first line of defense against the attack of seed predator. The most thoroughly examined plant serine proteinase inhibitors are found in the species of the families Leguminosae, Graminae, and Solanaceae. Leucaena leucocephala belongs to the family Leguminosae. It is widely used both as an ornamental tree as well as cattle food. We have constructed a three-dimensional model of a serine proteinase inhibitor from L. leucocephala seeds (LTI) complexed with trypsin. The model was built based on its comparative homology with soybean trypsin inhibitor (STI) using the program, MODELLER6. The quality of the model was assessed stereochemically by PROCHECK. LTI shows structural features characteristic of the Kunitz type trypsin inhibitor and shows 39% residue identity with STI. LTI consists of 172 amino acid residues and is characterized by two disulfide bridges. The protein is a dimer with the two chains being linked by a disulfide bridge. Despite the high similarity in the overall tertiary structure, significant differences exist at the active site between STI and LTI. The present study aims at analyzing these interactions based on the available amino acid sequences and structural data. We have also studied some functional sites such as phosphorylation, myristoylation, which can influence the inhibitory activity or complexation with other molecules. Some of the differences observed at the active site and functional sites can explain the unique features of LTI. PMID- 14741701 TI - Genomic organization, alternative splicing, and promoter analysis of human dynamin-like protein gene. AB - The human dynamin-like protein, HdynIV, has recently been cloned and shown to be involved in the formation and trafficking of coated vesicles. In particular, one of the HdynIV variant overexpressions has been suggested to contribute to the pathogenesis of brain tumors. In this paper, we report on the genomic organization of the human HdynIV gene. The gene was found to correspond to 20 exons of genomic sequence on human chromosome 12, distributed over 64kb of genomic DNA. The two exons, numbers 15 and 16, are subjected to differential splicing, generating four different transcripts of a perfect match to our recent report on the four different spliced HdynIV variants [DNA Cell Biol. 19 (2000) 189]. We have also characterized the 5(') regulatory region of the HdynIV gene in order to understand the molecular mechanisms regulating its expression. The transcriptional initiation site was identified by 5(')-RACE. The 5(')-flanking sequence of the HdynIV gene contains three GC boxes that concatenate Ap2- and Sp1 binding motifs, but that does not contain either the TATA or CAAT consensus sequence. A region between -140 and +92 contributed to high promoter activity. Deletion analysis demonstrated that the minimal promoter activity required the region of -110 to -100. Electrophoretic mobility shift assay demonstrated that a putative transcriptional factor bound to the region of -119 to -90. Site-directed mutagenesis analysis of this region revealed that nucleotides at -108 to -100 were essential for transactivation mediated by this transcriptional factor. In conclusion, we have characterized the minimal HdynIV promoter and shown that CTCCCAGCA (-108 to -100) sequence may act as a novel transcriptional element for regulating HdynIV gene expression. PMID- 14741702 TI - Neuropeptide Y promotes beta-cell replication via extracellular signal-regulated kinase activation. AB - AIMS/HYPOTHESIS: Previous studies have shown that neuropeptide Y (NPY) gene expression and release are increased in hyperphagic ob/ob mice and diabetic rats. Therefore, we hypothesized that orexigenic agent, NPY, has the effect on the obesity and diabetes. To elucidate the relationship, we have studied the regulatory role of NPY on islet cells. METHODS: Isolated islets were incubated with NPY or NPY Y1 receptor specific antagonist, BIBP3226. Proliferation, apoptosis, and Y1 receptor expression were identified by immunohistochemistry. We studied that ERK1/2 mediates the NPY pathway with PD98059 (MAP kinase inhibitor), wortmannin (phosphatidylinositol 3-kinase inhibitor), and BIM-1 (protein kinase C inhibitor). After NPY-treated islets were exposed to high glucose, insulin levels were detected. RESULTS: beta-Cell replication was enhanced in a dose-dependent manner, but without any changes on the other cells in islet. NPY Y1 receptors were expressed on islet and NPY induced phosphorylation of ERK1/2 rapidly and transiently. PD98059 (MAPK kinase inhibitor) and BIM-1 (protein kinase C inhibitor) inhibited activation of ERK1/2 by NPY, but wortmannin (phosphatidylinositol 3-kinase inhibitor) did not. Exposure of NPY-treated islets to high glucose showed the decreasing trend of insulin secretion. CONCLUSION/INTERPRETATION: Our data suggest that NPY promotes beta-cell replication via extracellular signal-regulated kinase activation and inhibits glucose-stimulated insulin secretion. PMID- 14741703 TI - HCV core protein-expressing DNA vaccine induces a strong class I-binding peptide DTH response in mice. AB - The hepatitis C virus (HCV) core protein-encoding sequence (HCcAg) is the most conserved gene in HCV genome and therefore may be useful to study broadly reacting T-cell epitopes. In this study BALB/c and C57BL/6 mice were immunized with a DNA based vaccine expressing the first 176 aa of HCcAg (pIDKCo). After i.m or i.p injection of pIDKCo in BALB/c mice, a detectable INF-gamma secreting response to the relevant class I-binding peptide DLMGYIPLVGA (P1) (aa 132-142) was detected suggesting the induction of HCcAg specific CD8(+) T-cell effectors. CD8(+) T-cell responses were also monitored in vivo by T-cell-mediated DTH reactions after subcutaneous injection of class I-binding viral peptide P1. pIDKCo induced a strong P1-specific DTH response in both i.m and i.p immunized mice. To evaluate the T-cell response induced by pIDKCo in C57BL/6 mice, an HCcAg epitope was predicted based upon it containing the H-2K(b) binding motif XXXXF/YXXL (DLMGYIPL (P2)). pIDKCo induced a strong P2-specific DTH response with similar kinetics of swelling response to that observed in BALB/c mice. Previously, it had been demonstrated that only activated and protective CD8(+) effector T cells could mediate a specific DTH in footpads of virally infected mice after local injection of viral class I-binding peptides. Hence, pIDKCo could prime a strong HCcAg-specific T-cell response in mice with the potential capacity to exert their specific effector functions in peripheral tissues. PMID- 14741704 TI - In situ surface electrochemical characterizations of Ti and Ti-6Al-4V alloy cultured with osteoblast-like cells. AB - The biocompatibility of metal implants is related to their surface electrochemical characterizations. The in situ growing process of osteoblast-like U-2 OS cells on polished Ti and Ti-6Al-4V alloy during 72h incubation was monitored using the electrochemical impedance spectroscopy (EIS) measurement technique. The results showed that the presence of cells on metals led to an increase in the impedance and polarization resistance (R(p)) of metals. The impedance and R(p) increased as the cells grew (i.e., from adhesion, spreading to proliferation period). A trace amount of V element released from Ti-6Al-4V alloy led to a lower R(p) with respect to Ti metal during cell culture. In this study, a satisfactory equivalent circuit simulating the electrochemical characterizations of Ti and Ti-6Al-4V alloy cultured with cells was proposed. The EIS measurement technique was applied successfully to monitor the in situ growing process of U-2 OS cells on Ti and Ti-6Al-4V alloy. PMID- 14741705 TI - PQQ glucose dehydrogenase with novel electron transfer ability. AB - PQQ glucose dehydrogenase from Acinetobacter calcoaceticus (GDH-B) is one of the most industrially attractive enzymes, as a sensor constituent for glucose sensing, because of its high catalytic activity and insensitivity to oxygen. We attempted to engineer GDH-B to enable electron transfer to the electrode in the absence of artificial electron mediator by mimicking the domain structure of the quinohemoprotein ethanol dehydrogenase (QH-EDH) from Comamonas testosteroni, which is composed of a PQQ-containing catalytic domain and a cytochrome c domain. We genetically fused the cytochrome c domain of QH-EDH to the C-terminal of GDH B. The constructed fusion protein showed not only intra-molecular electron transfer, between PQQ and heme of the cytochrome c domain, but also electron transfer from heme to the electrode, thereby allowing the construction of a direct electron transfer-type glucose sensor. PMID- 14741706 TI - The time-course of cyclic AMP signaling is critical for leukemia U-937 cell differentiation. AB - The regulation of the cAMP signaling is intimately involved in several cellular processes, including cell differentiation. Here, we provide strong evidence supporting that the time-course of cAMP signal is critical for leukemia U-937 cell differentiation. Three stimulating-cAMP agents were used to analyze the correlation between cAMP time-course and cell differentiation. All three agents denoted similar cAMP maximal responses in dose-response experiments. The kinetic of desensitization showed differential characteristics, while H2 receptor desensitized homologously without affecting PGE2 or forskolin effect, PGE2 response showed mixed desensitization characterized by a homologous initial phase followed by a heterologous phase. Regarding forskolin, long-term stimuli attenuated PGE2 and H2 agonist response without affecting adenylyl cyclase activity. In the absence of phosphodiesterase inhibitors, the three agents induced similar maximal cAMP levels after 5 min, but only that induced by the H2 agonist returned to basal levels. Consistent with this observation, H2 agonist was not able to induce U-937 cell maturation in contrast to PGE2 and forskolin, supporting the importance of time-course signaling in the determination of cell behavior. PMID- 14741707 TI - Existence of GPR40 functioning in a human breast cancer cell line, MCF-7. AB - GPR40, which has recently been identified as a G-protein-coupled cell-surface receptor for long-chain fatty acids, was assessed in a human breast cancer cell line (MCF-7). We detected GPR40 mRNA by RT-PCR and found that oleate and linoleate, but not palmitate or stearate, caused an increase in cellular Ca(2+) concentrations, which was partially blocked by the pertussis toxin (PTX) treatment. We examined the expression of GPR40 mRNA by quantitative RT-PCR in the relation to cell number. It was significantly increased at the beginning and at the end of cell proliferation. These results indicate the possibility that GPR40 for long-chain fatty acids may be involved in cellular function such as cell proliferation, providing a new perspective for the action of long-chain fatty acids on mammary epithelial cells. PMID- 14741708 TI - Spectroelectrochemistry of cytochrome P450cam. AB - The spectroelectrochemistry of camphor-bound cytochrome P450cam (P450cam) using gold electrodes is described. The electrodes were modified with either 4,4(') dithiodipyridin or sodium dithionite. Electrolysis of P450cam was carried out when the enzyme was in solution, while at the same time UV-visible absorption spectra were recorded. Reversible oxidation and reduction could be observed with both 4,4(')-dithiodipyridin and dithionite modified electrodes. A formal potential (E(0')) of -373mV vs Ag/AgCl 1M KCl was determined. The spectra of P450cam complexed with either carbon monoxide or metyrapone, both being inhibitors of P450 catalysis, clearly indicated that the protein retained its native state in the electrochemical cell during electrolysis. PMID- 14741709 TI - 15d-PGJ2 inhibits oxidized LDL-induced macrophage proliferation by inhibition of GM-CSF production via inactivation of NF-kappaB. AB - Macrophage-derived foam cells play an important role in atherosclerotic lesions. Oxidized low-density lipoprotein (Ox-LDL) induces macrophage proliferation via production of GM-CSF in vitro. This study investigated the effects of 15-deoxy Delta(12,14)-prostaglandin J(2) (15d-PGJ(2)), a natural ligand for peroxisome proliferator-activated receptor gamma, on macrophage proliferation. Mouse peritoneal macrophages and RAW264.7 cells were used for proliferation study and reporter gene assay, respectively. Twenty microgram per milliliter of Ox-LDL induced [3H]thymidine incorporation in mouse peritoneal macrophages, and 15d PGJ(2) inhibited Ox-LDL-induced [3H]thymidine incorporation in a dose-dependent manner. Ox-LDL increased GM-CSF release and GM-CSF mRNA expression, and activated GM-CSF gene promoter, all of which were prevented by 15d-PGJ(2) or 2-cyclopenten 1-one, a cyclopentenone ring of 15d-PGJ(2). The suppression of GM-CSF promoter activity by 15d-PGJ(2) and 2-cyclopenten-1-one was mediated through reduction of NF-kappaB binding to GM-CSF promoter. These results suggest that 15d-PGJ(2) inhibits Ox-LDL-induced macrophage proliferation through suppression of GM-CSF production via NF-kappaB inactivation. PMID- 14741710 TI - Biotin enhances ATP synthesis in pancreatic islets of the rat, resulting in reinforcement of glucose-induced insulin secretion. AB - Previous studies showed that biotin enhanced glucose-induced insulin secretion. Changes in the cytosolic ATP/ADP ratio in the pancreatic islets participate in the regulation of insulin secretion by glucose. In the present study we investigated whether biotin regulates the cytosolic ATP/ADP ratio in glucose stimulated islets. When islets were stimulated with glucose plus biotin, the ATP/ADP ratio increased to approximately 160% of the ATP/ADP ratio in islets stimulated with glucose alone. The rate of glucose oxidation, assessed by CO(2) production, was also about 2-fold higher in islets treated with biotin. These increasing effects of biotin were proportional to the effects seen in insulin secretion. There are no previous reports of vitamins, such as biotin, directly affecting ATP synthesis. Our data indicate that biotin enhances ATP synthesis in islets following the increased rate of substrate oxidation in mitochondria and that, as a consequence of these events, glucose-induced insulin release is reinforced by biotin. PMID- 14741711 TI - Pervanadate-induced reverse translocation and tyrosine phosphorylation of phorbol ester-stimulated protein kinase C betaII are mediated by Src-family tyrosine kinases in porcine neutrophils. AB - Protein kinase C (PKC), upon activation, translocates from the cytosol to the plasma membrane. Phorbol 12-myristate 13-acetate (PMA), a potent PKC activator, is known to induce irreversible translocation of PKC to the plasma membrane, in contrast to the reversible translocation resulting from physiological stimuli and subsequent rapid return to the cytosol (reverse translocation). However, we have previously shown that tyrosine phosphatase (PTPase) inhibitors induce reverse translocation of PMA-stimulated PKCbetaII in porcine polymorphonuclear leukocytes (PMNs). In the present study, we showed that pervanadate, a potent PTPase inhibitor, also induces tyrosine phosphorylation of PMA-stimulated PKCbetaII in porcine PMNs. Furthermore, PP2, a specific inhibitor of Src-family tyrosine kinases (PTKs), was found to inhibit both pervanadate-induced reverse translocation and tyrosine phosphorylation of PMA-stimulated PKCbetaII, suggesting that these two pervanadate-induced responses are mediated by Src family PTKs. Our findings provide novel insight into the relationship between the subcellular localization and tyrosine phosphorylation of PKC. PMID- 14741712 TI - Identification and characteristic analysis of the ampC gene encoding beta lactamase from Vibrio fischeri. AB - Vibrio fischeri ATCC 7744 is an ampicillin resistant (Amp(r)) marine luminous bacterium. The MIC test indicates that V. fischeri is highly resistant to penicillins, and susceptible to cephalosporins. V. fischeri ampC gene was cloned and identified. Nucleotide sequence of an unidentified ufo gene and the ampC, ppiB genes (GenBank Accession No. AY438037) has been determined; whereas the ampC gene encodes the beta-lactamase (AmpC) and the ppiB gene encodes the peptidyl prolyl cis-trans isomerase B. Alignment and comparison show that V. fischeri beta lactamase is homologous to the related species'. The specific amino acid residues STFK (62nd to 65th), SDN (122nd to 124th), and D (155th) located 34 residues downstream from the SDN loop of the class A beta-lactamases are highly conserved, but the KTG is not found. V. fischeri ampC gene encoding beta-lactamase has a calculated M(r) 31,181 and comprises 283 amino acid residues (pI 5.35). There is a signal peptide of 18 amino acid residues MKIKPFLFGLIVLANNAI in the pro-beta lactamase, which functioned for secretion; thus, the matured protein only has M(r) 29,197 and comprises 265 amino acid residues (pI 4.95). SDS-PAGE and the beta-lactamase functional assays elicit that the M(r) of the beta-lactamases are close to 29kDa. IEF and the beta-lactamase functional assays show that the beta lactamases' pI are close to 4.8 as predicted. The results elucidate that V. fischeri ampC gene and the cloned ampC gene in Escherichia coli are the same one. The gene order of the ampC and the related genes is -ufo-(P*-intern)-ampC-ppiB--> (P*-intern: intern promoter for sub-regulation), whereas the P*-intern promoter displays the function to lead the ampC gene's expression for stress response. PMID- 14741713 TI - Overexpression of Ssl2p confers resistance to adriamycin and actinomycin D in Saccharomyces cerevisiae. AB - Adriamycin is one of the most active anticancer drugs but the development of resistance to this drug hampers its efficacy. In an effort to identify novel genes that confer resistance to adriamycin, we introduced a yeast genomic library into Saccharomyces cerevisiae and selected transformants that grew in the presence of a normally toxic concentration of adriamycin. Detailed examination of a plasmid recovered from these transformants revealed that overexpression of the gene for Ssl2p rendered yeast cells resistant to adriamycin. Ssl2p is a protein that is involved in the initiation of transcription and in DNA repair. Overexpression of Ssl2p did not confer resistance to aclarubicin, an anthracycline anticancer drug, which, like adriamycin, is intercalated into DNA. Both adriamycin and aclarubicin inhibit topoisomerase II and, thus, topoisomerase II might not be a major factor in the acquired resistance to adriamycin that results from overexpression of Ssl2p. We tested several other compounds but the only one to which Ssl2p-overexpressing cells were cross-resistant was actinomycin D. Mammalian cells that overexpress P-glycoprotein, which is a transmembrane protein that is involved in the efflux of certain drugs, are resistant to both adriamycin and actinomycin D but not to aclarubicin. However, overexpression of Ssl2p had little or no effect on the intracellular accumulation of adriamycin. Our results suggest that a novel mechanism might be involved in the sensitivity of yeast to both adriamycin and actinomycin D. PMID- 14741714 TI - Role of Na+-Ca2+ exchanger in myocardial ischemia/reperfusion injury: evaluation using a heterozygous Na+-Ca2+ exchanger knockout mouse model. AB - We used Na(+)-Ca(2+) exchanger (NCX) knockout mice to evaluate the effects of NCX in cardiac function and the infarct size after ischemia/reperfusion injury. The contractile function in NCX KO mice hearts was significantly better than that in wild type (WT) mice hearts after ischemia/reperfusion and the infarct size was significantly small in NCX KO mice hearts compared with that in WT mice hearts. NCX is critically involved in the development of ischemia/reperfusion-induced myocardial injury and therefore the inhibition of NCX function may contribute to cardioprotection against ischemia/reperfusion injury. PMID- 14741715 TI - Profiling the allosteric response of an engineered beta-galactosidase to its effector, anti-HIV antibody. AB - Escherichia coli beta-galactosidase responds enzymatically to antiviral antibodies when a viral antigenic peptide, acting as receptor, is conveniently displayed in the vicinity of the active site. The allosteric response of a beta galactosidase molecular sensor containing a B-cell epitope from HIV has been finely dissected upon binding of an effector monoclonal antibody, within a wide range of standard concentrations of both enzyme and substrate. The topography of the enzymatic activation reveals a wide set of conditions in which the enzymatic response renders a signal over threefold the background, that is suitable for analytical biosensing. Moreover, at discrete enzyme-substrate coordinates, the effector antibody promotes an enhanced activation factor up to fivefold. The insertion of the 37-mer viral peptide between beta-galactosidase residues 795 and 796 is observed as inducer of the structural flexibility required for molecular sensing, whose dynamics and efficiency are intimately associated with the concentrations of enzyme and substrate, the two partners in the signal transduction event. PMID- 14741716 TI - Structure-activity function for binding and signaling in CHO-K1 and COS-7 cells expressing the cholecystokinin A receptor. AB - Key amino acids of the cholecystokinin (CCK) peptide for receptor binding are sulfated Y27, W30, D32, and F33-NH(2). Three-dimensional modeling showed that the CCK-A receptor (CCK-AR) antagonist devazepide penetrated into the transmembrane (TM) domains, whereas CCK was placed on the surface of the CCK-AR. Four types of rat CCK-AR cDNAs were transfected into CHO-K1 and COS-7 cells: normal CCK-AR cDNA transfected cells (wild type, WT); K120 substituted with V; K130V; and R352V. Binding of [3H]CCK-8 was observed in WT and K130V, but not in K120V and R352V. CCK caused Ca(2+) spiking in WT and K130V, whereas K120V and R352V had no effect. Three chimeras including the CCK-AR/3ibeta2 adrenergic receptor (beta2AR), 3Nibeta2AR, and 3Cibeta2AR were constructed. Two groups of point mutations in the CCK-AR3i were also made: Y252V, S274V, S281V, and S289V (non-phospho-acceptor Y or S); S260V, S264V, S271V, and S275V (phospho-acceptor S). WT and CCK AR/3Cibeta2AR increased [Ca(2+)](i) in response to CCK; 3Nibeta2AR was vice versa. CCK failed to increase [IP(3)] in phospho-acceptor S to V without affecting binding. Non-phospho-acceptor S or Y to V showed normal response. Thus, Lys120 outside the TM2 and Arg352 outside the TM6 of the CCK-AR are amino acids interacting with Tyr[SO(3)H]27 and Asp32 of the CCK peptide for binding. Phospho acceptor Ser groups in the CCK-AR 3Ni are amino acids for initiating cell signaling. PMID- 14741717 TI - Chronological characterization of diabetes development in male Spontaneously Diabetic Torii rats. AB - To characterize the underlying mechanisms of diabetes development in males of the Spontaneously Diabetic Torii (SDT) rat, a novel spontaneous model for diabetes, we chronologically examined them, focusing on their diabetic features and the pathological changes in the pancreatic islets. Male SDT rats exhibited glucose intolerance with impaired insulin secretion after 14 weeks and developed diabetes with remarkable hyperglycemia and marked hypoinsulinemia after 20 weeks. At prediabetic stage (10-20 weeks), they were normoglycemic, but had significantly lower insulin levels of plasma and pancreas than the normal rats. Their beta-cell volume was already smaller significantly at 10 weeks than that of normal rats. The primary changes of the pancreatic islets were microvascular events such as congestion and hemorrhage at 8-10 weeks. Thereafter, the SDT rat islets were affected with inflammation and progressive fibrosis (at 10-20 weeks), and eventually atrophied with a loss of beta-cells (at 38 weeks). These results indicate that the male SDT rats develop spontaneous diabetes with an absolute decrease in the insulin secretory capacity of the islets. PMID- 14741718 TI - Single-channel gating and regulation of human L-type calcium channels in cardiomyocytes of transgenic mice. AB - Overexpression of human cardiac L-type Ca(2+) channel pores (hCa(v)1.2) in mice causes heart failure. Earlier studies showed Ca(v)1.2-mRNA increase by 2.8-fold, but whole-cell current density enhancement by >> Tn-glycopeptides (M.W. <3.0 x 10(3))/Talpha monomer > monovalent P/S > Tn monomer and GalNAc > tri-antennary II > Gal >> Man and Glc (inactive). These findings give evidence for the binding of this lectin to dense cell surface T, Tn and I/II glycoconjugates and should facilitate future usage of this lectin in biotechnological and medical applications. PMID- 14741735 TI - Metabolism of cyclic ADP-ribose: Zinc is an endogenous modulator of the cyclase/NAD glycohydrolase ratio of a CD38-like enzyme from human seminal fluid. AB - CD38, a bifunctional enzyme capable of both synthesis and hydrolysis of the second messenger cyclic ADP-ribose (cADPR). Using the natural substrate of the enzyme, NAD+, the ratio of ADP-ribosyl cyclase/NAD glycohydrolase of CD38 is about 1/100. Here we describe that human seminal fluid contain a soluble CD38 like enzyme with an apparent M.W. of 49 kDa. When purified this enzyme has a cyclase/NAD glycohydrolase ratio of about 1/120. However, the in situ cyclase/NAD glycohydrolase ratio measured in seminal plasma approaches 1/1. We also found that physiological concentrations of zinc present in the seminal fluid, in the range of 0.6 to 4 mM, are responsible for the modulation of the cyclase/NAD glycohydrolase ratio. This new information indicates that the cyclase/NAD glycohydrolase ratio can be modified in vivo. PMID- 14741736 TI - Inhibition of human tumor cell proliferation by novel anthraquinones from daylilies. AB - Daylilies (Hemerocallis) are used medicinally in eastern Asia and extracts of the plant had been shown to inhibit cell proliferation and induce cancer cells to undergo differentiation. In our studies of the constituents of Hemerocallis fulva var. 'Kwanzo' roots, we isolated a series of new [kwanzoquinones A (1), B (2), C (4), D (5), E (6), F (7), G (9)] and known [2-hydroxychrysophanol (3) and rhein (8)] anthraquinones. These compounds were tested in order to determine their potential roles as cancer cell growth inhibitors. Kwanzoquinones A-C and E, kwanzoquinone A and B monoacetates (1a and 2a), 2-hydroxychrysophanol, and rhein inhibited the proliferation of human breast, CNS, colon, and lung cancer cells with GI50 values between 1.8 to 21.1 microg/mL. However, upon exposure of the cancer cells to the GI50 concentrations of the bioactive anthraquinones, most of the cancer cell lines exhibited higher than anticipated levels of cell viability. Co-incubation of the anthraquinones with vitamins C and E increased the viability of breast cancer cells. In contrast, vitamins C and E potentiated the cytotoxic effects of the anthraquinones against the colon cancer cells. None of the anthraquinones inhibited the activity of topoisomerase. PMID- 14741737 TI - Chronic leptin treatment enhances insulin-stimulated glucose disposal in skeletal muscle of high-fat fed rodents. AB - The aim of this investigation was to evaluate if chronic leptin administration corrects high fat diet-induced skeletal muscle insulin resistance, in part, by enhancing rates of glucose disposal and if the improvements are accounted for by alterations in components of the insulin-signaling cascade. Sprague-Dawley rats consumed normal (CON) or high fat diets for three months. After the dietary lead in, the high fat diet group was further subdivided into high fat (HF) and high fat, leptin treated (HF-LEP) animals. HF-LEP animals were injected twice daily with leptin (5 mg/100 g body weight) for 10 days, while the CON and HF animals were injected with vehicle. Following the treatment periods, all animals were prepared for and subjected to hind limb perfusion. The high fat diet decreased rates of insulin-stimulated skeletal muscle glucose uptake and glycogen synthesis in the red gastrocnemius (RG), but did not affect glycogen synthase activity, rates of glucose oxidation or nonoxidative disposal of glucose. Of interest, IRS 1-associated PI3-K activity and total GLUT4 protein concentration were reduced in the RG of the high fat-fed animals. Leptin treatment increased rates of insulin stimulated glucose uptake and glucose oxidation, and normalized rates of glycogen synthesis. Leptin appeared to mediate these effects by normalizing insulin stimulated PI3-K activation and GLUT4 protein concentration in the RG. Collectively, these data suggest that chronic leptin treatment reverses the effects of a high fat diet thereby allowing the insulin signaling cascade and glucose transport effector system to be fully activated which in turn affects the amount of glucose that is transported across the plasma membrane and made available for glycogen synthesis. PMID- 14741738 TI - The specific p38 mitogen-activated protein kinase pathway inhibitor FR167653 keeps insulitis benign in nonobese diabetic mice. AB - The p38 mitogen-activated protein kinase (MAPK) pathway is important in Th1 immunity, macrophage activation, and apoptosis. Since they may be associated with beta-cell destruction during the development of type 1 diabetes, we investigated the role of the p38 MAPK pathway in female nonobese diabetic (NOD) mice. Phosphorylated p38 MAPK was observed immunohistochemically in CD4+ cells that had infiltrated into the islets and part of beta-cells, increasing in proportion to the severity of insulitis. Continuous oral administration of 0.08% FR167653, a specific p38 MAPK pathway inhibitor, significantly reduced the ex vivo production of interferon-gamma by splenic Th1 cells without affecting interleukin-4 production by Th2 cells. FR167653 administration from 4-30 weeks of age prevented NOD mice from developing diabetes without affecting the severity of insulitis. Treatment with FR167653 after insulitis had developed (i.e. from 10-30 weeks of age) also prevented diabetes, further suggesting that treatment with the p38 MAPK pathway inhibitor keeps insulitis benign in NOD mice, partly by inhibiting Th1 immunity. These findings suggest that p38 MAPK is a key mediator that switches insulitis from benign to destructive in the development of type 1 diabetes. PMID- 14741739 TI - Cellular activity and signaling induced by osteoprotegerin in osteoclasts: involvement of receptor activator of nuclear factor kappaB ligand and MAPK. AB - Osteoprotegerin (OPG) is a decoy receptor for receptor activator of nuclear factor kappaB ligand (RANKL), an inducer of osteoclastogenesis via its receptor RANK. We recently demonstrated that OPG also exerts a direct effect in osteoclasts by regulating protease expression. Herein, we showed that OPG-induced pro-matrix metalloproteinase-9 activity was abolished by ras/MAPK inhibitors in purified osteoclasts. OPG induced the phosphorylation of p38 and ERK1/2 in RAW264.7 cells. Only p38 activation was totally abolished by a blocking anti RANKL antibody or an excess of RANKL. Surface plasmon resonance experiments revealed that RANK, RANKL and OPG are able to form a tertiary complex. These results suggested a potential formation of a tertiary complex RANK-RANKL-OPG on osteoclasts. Thus, OPG is not only a soluble decoy receptor for RANKL but must be also considered as a direct effector of osteoclast functions. PMID- 14741740 TI - Hepatocyte growth factor/scatter factor suppresses TNF-alpha-induced E-selectin expression in human umbilical vein endothelial cells. AB - Induction of E-selectin on endothelial cell surface initiates leukocyte adhesion and subsequent migration into the subendothelium. Here, we tested the effect of hepatocyte growth factor (HGF) on inflammatory cytokine-induced expression of E selectin and consequent leukocyte-endothelial cell interaction using human umbilical vein endothelial cells (HUVEC). Prior treatment of HUVEC with HGF significantly attenuated the tumor necrosis factor (TNF)-alpha-induced E-selectin protein, adhesion of HL60 cells to HUVEC and E-selectin mRNA expression in a dose dependent manner, while HGF itself did not exert any effects. The HGF effects on the mRNA expression were inhibited in the presence of N(G)-nitro-L-arginine methyl ester (L-NAME), a nitric oxide synthase (NOS) inhibitor, which also abolished HGF-stimulated eNOS activity. These results suggest HGF plays cardiovascular protective functions mediated, at least in part, through nitric oxide-dependent suppression of inflammatory cytokine-induced E-selectin expression and subsequent tethering of leukocytes to endothelial cells. PMID- 14741741 TI - Endothelin-1 promotes phosphorylation of CREB transcription factor in primary cultures of neonatal rat cardiac myocytes: implications for the regulation of c jun expression. AB - Cardiac myocyte hypertrophy is associated with an increase in expression of immediate early genes (e.g. c-jun) via activation of pre-existing transcription factors. The activity of CREB transcription factor is regulated through phosphorylation of Ser-133 by one of several protein kinases (e.g. protein kinase A (PKA), p90 ribosomal S6 kinases (RSKs) and the related kinase, MSK1). A cell permeable form of cAMP, hypertrophic agonists (endothelin-1 (ET-1), phenylephrine (PE)) and hyperosmotic shock all promoted phosphorylation of CREB(Ser-133) in rat neonatal cardiac myocytes. The response to endothelin-1 required the extracellular signal-regulated kinase cascade which stimulates both RSKs and MSK1. Phosphorylation of CREB(Ser-133) in response to ET-1 was not associated with any increase in DNA binding to a consensus cAMP-response element (CRE). The rat c-jun promoter contains elements which may bind either c-Jun/ATF2 or CREB/ATF1 dimers. Using extracts from rat cardiac myocytes, we identified at least two complexes which bind to the most proximal of these elements, one of which contained CREB and the other c-Jun. Thus, phosphorylation and activation of CREB in cardiac myocytes may be effected by a range of different stimuli to influence the expression of immediate early genes such as c-jun. PMID- 14741742 TI - The intra-mitochondrial cytochrome c distribution varies correlated to the formation of a complex between VDAC and the adenine nucleotide translocase: this affects Bax-dependent cytochrome c release. AB - The mechanism of Bax-dependent cytochrome c release is still controversial and may also depend on the actual localisation of cytochrome C: (i) we studied the distribution of cytochrome c in sub-fractions of rat kidney mitochondria and found that 10-20% of the total cytochrome c was associated at the peripheral inner membrane and to some extent organised in the contact sites. (ii) Cytochrome c concentrations in the contact site fractions varied related to surface bound hexokinase activity. It decreased upon reduction of contact sites by glycerol or specific dissociation of the VDAC-ANT complexes by bongkrekate, whereas it increased upon induction of contacts by dextran or association of VDAC-ANT complexes by atractyloside. (iii) The outer membrane pore (VDAC) acquires high capacity for hexokinase binding by interacting with the ANT. Thus, surface attached hexokinase protein indicated the frequency of VDAC-ANT complexes and the correlation between hexokinase activity and cytochrome c suggested association of the latter to the complexes. (iv) Substances affecting exclusively the structure of either hexokinase (glucose-6P) or cytochrome c (borate) led to a decrease only of the effected protein without changing the concentration of other contact site constituents. (v) Hexokinase was furthermore used as a tool to isolate the contact site forming complex of outer membrane VDAC and inner membrane ANT from Triton-dissolved membranes. Cytochrome c remained attached to the hexokinase VDAC ANT complexes that were reconstituted in phospholipid vesicles. (vi) The vesicles were loaded with malate and BaxDeltaC released the endogenous cytochrome c from the reconstituted complexes without forming unspecific pores for malate. BaxDeltaC targeted a cytochrome c fraction associated at the VDAC-ANT complex. The cytochrome c organisation was dependent on the actual structure of VDAC and ANT. Thus, the BaxDeltaC effect was suppressed either by hexokinase utilising glucose and ATP or by bongkrekic acid both influencing the pore and ANT structure. PMID- 14741743 TI - Toxicity of ethanol and acetaldehyde in hepatocytes treated with ursodeoxycholic or tauroursodeoxycholic acid. AB - In hepatocytes ethanol (EtOH) is metabolized to acetaldehyde and to acetate. Ursodeoxycholic acid (UDCA) and tauroursodeoxycholic acid (TUDCA) are said to protect the liver against alcohol. We investigated the influence of ethanol and acetaldehyde on alcohol dehydrogenase (ADH)-containing human hepatoma cells (SK Hep-1) and the protective effects of UDCA and TUDCA (0.01 and 0.1 mM). Cells were incubated with 100 and 200 mM ethanol, concentrations in a heavy drinker, or acetaldehyde. Treatment with acetaldehyde or ethanol resulted in a decrease of metabolic activity and viability of hepatocytes and an increase of cell membrane permeability. During simultaneous incubation with bile acids, the metabolic activity was better preserved by UDCA than by TUDCA. Due to its more polar character, acetaldehyde mostly damaged the superficial, more polar domain of the membrane. TUDCA reduced this effect, UDCA was less effective. Damage caused by ethanol was smaller and predominantly at the more apolar site of the cell membrane. In contrast, preincubation with TUDCA or UDCA strongly decreased metabolic activity and cell viability and led to an appreciable increase of membrane permeability. TUDCA and UDCA only in rather high concentrations reduce ethanol and acetaldehyde-induced toxicity in a different way, when incubated simultaneously with hepatocytes. In contrast, preincubation with bile acids intensified cell damage. Therefore, the protective effect of UDCA or TUDCA in alcohol- or acetaldehyde-treated SK-Hep-1 cells remains dubious. PMID- 14741744 TI - Identification of major proteins in the lipid droplet-enriched fraction isolated from the human hepatocyte cell line HuH7. AB - Recent studies have revealed the presence of intracellular lipid droplets in wide variety of species. In mammalian cells, there exist proteins specifically localize in lipid droplets. However, the protein profile in the droplet remains yet to be clarified. In this study, a fraction enriched with lipid droplets was isolated from a human hepatocyte cell line HuH7 using sucrose density gradient centrifugation, and 17 major proteins in the fraction were identified using nano LC-MS/MS techniques. Adipose differentiation-related protein (ADRP) was the most abundant protein in the fraction. The secondary abundant proteins were identified to be acyl-CoA synthetase 3 (ACS3) and 17beta-hydroxysteroid dehydrogenase 11 (17betaHSD11). Included in the identified proteins were five lipid-metabolizing enzymes as well as two lipid droplet-specific proteins. When HuH7 cell lysate was fractionated by a density gradient, most of 17betaHSD11 was found in the droplet enriched fraction. In immunocytochemical analysis, 17betaHSD11 showed ring-shaped images which overlapped with those for ADRP. These results suggest that a specific set of proteins is enriched in the lipid droplet-enriched fraction and that 17betaHSD11 localizes specifically in the fraction. PMID- 14741745 TI - Phosphorylation of MAP kinase-like proteins mediate the response of the halotolerant alga Dunaliella viridis to hypertonic shock. AB - The microalga Dunaliella viridis has the ability to adapt to a variety of environmental stresses including osmotic and thermal shocks, UV irradiation and nitrogen starvation. Lacking a rigid cell wall, Dunaliella provides an excellent model to study stress signaling in eukaryotic unicellular organisms. When exposed to hyperosmotic stress, UV irradiation or high temperature, a 57-kDa protein is recognized by antibodies specific to mammalian p38, to its yeast homologue Hog1, and to the phospho-p38 MAP kinase motif. This 57-kDa protein appears to be both up-regulated and phosphorylated. Three other proteins (50, 45, 43 kDa) were transiently phosphorylated under stress conditions as detected with an antibody specific to the mammalian phospho c-Jun N-terminal kinase (JNK) motif. Treatment with specific inhibitors of p38 MAP kinase (SB203580) and JNK (SP600125) activities markedly impaired the adaptation of Dunaliella to osmotic stress. From an evolutionary standpoint, these data strongly suggest that MAP kinase signaling pathways, other than ERK, were already operating in the common ancestor of plant and animal kingdoms, probably as early as 1400 million years ago. PMID- 14741747 TI - Making connections: the development of mesencephalic dopaminergic neurons. AB - The disorders of two adjacent sets of mesencephalic dopaminergic (MDNs) are associated with two significant health problems: Parkinson's disease and drug addiction. Because of this, a great deal of research has focused on understanding the growth, development and maintenance of MDNs. Many transcription factors and signaling pathways are known to be required for normal MDNs formation, but a unified model of MDN development is still unclear. The long-term goal is to design therapeutic strategies to: (i) nurture and/or heal endogenous MDNs, (ii) replace the affected tissue with exogenous MDNs from in vitro cultivated stem cells and (iii) restore normal connectivity. Recent developmental biology studies show great promise in understanding how MDNs develop both in vivo and in vitro. This information has great therapeutic value and may provide insight into how environmental and genetic factors increase vulnerability to addiction. PMID- 14741748 TI - Consequences of prenatal cocaine exposure in nonhuman primates. AB - The extent to which cocaine abuse by pregnant women can affect development of their offspring remains a matter of significant debate. In large part, this is due to difficulties in accurate determination of the type, dose, and pattern of cocaine administration by drug abusing women as well as to difficulties in controlling for a wide range of potentially confounding variables, such as other drugs used, race, socioeconomic status, and level of prenatal care. On this background, examination of the effects of prenatal cocaine exposure in highly controlled nonhuman primate models represents an important complement to the human research. The present review summarizes the data obtained in several different rhesus monkey models of cocaine exposure in utero. These data demonstrate the potential of prenatal cocaine exposure to interfere with structural and biochemical development of the brain leading to behavioral deficits at birth and/or during adulthood. However, the differences in the outcomes between individual models also suggest that the specific types and severity of cocaine effects are likely dependent on the route, dose, gestational period, and daily pattern of administration. PMID- 14741749 TI - Drugs of abuse that cause developing neurons to commit suicide. AB - When neuronal activity is abnormally suppressed during the developmental period of synaptogenesis, the timing and sequence of synaptic connections is disrupted, and this causes nerve cells to receive an internal signal to commit suicide, a form of cell death known as "apoptosis". By altering glutamate and GABA transmission alcohol suppresses neuronal activity, causing millions of nerve cells to commit suicide in the developing brain. This proapoptotic effect of alcohol provides a likely explanation for the diminished brain size and lifelong neurobehavioral disturbances associated with the human fetal alcohol syndrome. These findings have public health significance, not only in relation to fetal alcohol syndrome, but also in relation to several other drugs of abuse and various drugs used in obstetric and pediatric medicine, because these additional drugs (e.g. phencyclidine, ketamine, benzodiazepines, barbiturates) also suppress neuronal activity and drive developing neurons to commit suicide. PMID- 14741750 TI - Does drug abuse beget drug abuse? Behavioral analysis of addiction liability in animal models of prenatal drug exposure. AB - Prenatal exposure to drugs of abuse is the single largest preventable cause of developmental compromise of American children today. In the clinical population, it is difficult to determine the independent effects of gestational exposure to a single drug on brain development, in part due to the confounding effects of additional risk factors that are encountered in the substance-abusing population. The enormous clinical and societal problem of gestational toxicity of drugs of abuse, both legal and illegal, has driven the need to develop and investigate animal models of gestational drug exposure in which these variables can be controlled. More specifically, as clinical data are gathered suggesting an increased liability to substance abuse among children of drug-abusing mothers, a mechanistic understanding of the lasting effects of early drug exposure on the developing brain and the behavioral repertoire of the developing animal is crucial. In this review we summarize experimental animal research that investigates the role of drug exposure in utero on the functional development of specific brain circuits that are involved in the reinforcing effects of drugs of abuse, and on the behaviors that are mediated by these brain reward systems. PMID- 14741751 TI - Regional differences in cortical dendrite morphology following in utero exposure to cocaine. AB - In utero exposure to cocaine has been shown to affect dopaminergic populations of developing neurons in the central nervous system (CNS). To determine if this was a regionally specific effect or the result of a global phenomenon, we used a Golgi-Cox analysis to measure several parameters of neuronal development in murine neurons from frontal cortex, a region of the cortex containing monoamine innervation, and somatosensory cortex, a monoamine sparse part of the cortex. Results of these analyses show that in utero exposure to cocaine affects total dendrite length in histotypical layers III and IV and dendritic volume in layer III of the frontal cortex. These effects are not present in the somatosensory cortex. PMID- 14741752 TI - Prenatal exposure to cocaine decreases adenylyl cyclase activity in embryonic mouse striatum. AB - Adenylyl cyclase activity was measured in the striatum of naive mice as a function of age and in mice exposed in utero to cocaine. In naive Swiss-Webster mice, basal and forskolin-stimulated adenylyl cyclase activity increased gradually from embryonic day 13 (E13) until 2-3 weeks of age when activity peaked before decreasing slightly to adult levels. The ability of the dopamine D1 receptor agonist, SKF 82958, to stimulate adenylyl cyclase activity also increased in magnitude until P15. In a separate study, pregnant Swiss-Webster mice were injected twice daily with cocaine (15 mg/kg, s.c.) or an equal volume of saline from E10 to E17. Adenylyl cyclase activity was measured in the striatum of E18 embryos. Basal adenylyl cyclase activity was significantly reduced following prenatal exposure to cocaine. Likewise, the ability of forskolin or SKF 82958 to stimulate adenylyl cyclase was attenuated following cocaine exposure. DeltaFosB was not induced, contrary to what is seen in adult mice. These results demonstrate a functional change in a critical signal transduction pathway following chronic in utero exposure to cocaine that might have profound effects of the development of the brain. Alterations in the cAMP system may underlie some of the deficits seen in humans exposed in utero to cocaine. PMID- 14741753 TI - Prenatal cocaine administration increases glutathione and alpha-tocopherol oxidation in fetal rat brain. AB - Recent findings suggest that prenatal cocaine exposure results in significant attenuation of uterine and placental blood flow. The extent of blood flow reduction to fetuses positively correlates with reductions in glial-derived neurotrophic factor (GDNF) and dopamine (DA). However, whether such changes in uterine blood flow are sufficient to induce oxidative stress have yet to be determined. In the following experiments, the impact of prenatal cocaine exposure on fetal brain levels of the endogenous antioxidant glutathione (GSH and its oxidized form GSSG) or the exogenous antioxidant alpha-tocopherol (alpha-T and its oxidized quinone form) was investigated. It was hypothesized that cocaine exposure would result in greater oxidation of both GSH and alpha-T. Results indicated that a single injection of cocaine to a drug-naive pregnant dam results in significant (-16.38%) reductions in the levels of GSH. GSSG can be either raised or reduced as a result of fetal uterine position: fetuses at the ovarian extremes show significant increases in GSSG in response to cocaine (+64.73%), whereas cervically situated fetuses show decreased GSSG (-47.91%). Additionally, cocaine significantly decreased the levels of alpha-T (-15.9%) and increased the levels of its oxidative product alpha-Tquinone (alpha-Tq, +34.05%). Levels of alpha-T were not affected by fetal uterine position. These data collectively suggest that cocaine exposure increases the utilization of both endogenous and exogenous anti-oxidants in the fetal rat brain. Along with previous studies, these data support the hypothesis that cocaine-induced vasoconstriction results in oxidative stress in the gestating fetus. PMID- 14741754 TI - Impaired sustained attention and altered reactivity to errors in an animal model of prenatal cocaine exposure. AB - Although correlations have been reported between maternal cocaine use and impaired attention in exposed children, interpretation of these findings is complicated by the many risk factors that differentiate cocaine-exposed children from SES-matched controls. For this reason, the present dose-response study (0, 0.5, 1.0, or 3.0 mg/kg cocaine HCl) was designed to explore the effect of prenatal cocaine exposure on visual attention in a rodent model, using an intravenous injection protocol that closely mimics the pharmacokinetic profile and physiological effects of human recreational cocaine use. In adulthood, animals were tested on an attention task in which the duration, location, and onset time of a brief visual cue varied randomly between trials. The 3.0 mg/kg exposed males committed significantly more omission errors than control males during the final 1/3 of each testing session, specifically on trials that followed an error, which implicates impaired sustained attention and increased reactivity to committing an error. During the final 1/3 of each testing session, the 0.5 and 1.0 mg/kg exposed females took longer to enter the testing alcove at trial onset, and failed to enter the alcove more frequently than control females. Because these effects were not seen in other tasks of similar duration and reinforcement density, these findings suggest an impairment of sustained attention. This inference is supported by the finding that the increase in omission errors in the final block of trials in each daily session (relative to earlier in the session) was significantly greater for the 1.0 mg/kg females than for controls, a trend also seen for the 0.5 mg/kg group. Unlike the cocaine exposed males, who remain engaged in the task when attention is waning, the cocaine-exposed females appear to opt for another strategy; namely, refusing to participate when their ability to sustain attention is surpassed. PMID- 14741755 TI - Altered cocaine-induced behavioral sensitization in adult mice exposed to cocaine in utero. AB - Behavioral sensitization induced by psychostimulants is characterized by increased locomotion and stereotypy and may reflect aspects of neuronal adaptations underlying drug addiction in humans. To study the developmental contributions to addictive behaviors, we measured behavioral responses in adult offspring to a cocaine sensitization paradigm following prenatal cocaine exposure. Pregnant Swiss-Webster (SW) mice were injected twice daily from embryonic days 8 to 17 (E8-E17, inclusive) with cocaine (20 or 40 mg/kg/day; COC20 and COC40, respectively), or saline vehicle (SAL and SPF40) subcutaneously (s.c.). A nutritional control group of dams were 'pair-fed' with COC40 dams (SPF40). P120 male offspring from each prenatal treatment group were assigned to a behavioral sensitization group and injected with cocaine (15 mg/kg) or saline intraperitoneally (i.p.) every other day for seven doses. Locomotor activity and stereotypy were measured during habituation, cocaine initiation, and following a cocaine challenge 21 days after the last initiation injection. As expected, animals demonstrated significantly more locomotion and stereotypic behavior following acute and recurrent injection of cocaine compared to saline-injected animals. However, for each prenatal treatment group, cocaine-sensitized animals showed unique temporal profiles for the increase in locomotor sensitization and stereotypy over the course of the sensitization protocol. Two features that distinguished the altered behavioral progression of prenatally cocaine-exposed animals (COC40) from control (SAL) animals included blunted augmentation of locomotion and enhanced patterns of stereotypic behavior. These findings provide evidence that the behavioral activating effects of cocaine in adult animals are altered following exposure to cocaine in utero. PMID- 14741756 TI - c-fos and cleaved caspase-3 expression after perinatal exposure to ethanol, cocaine, or the combination of both drugs. AB - Poly-drug abuse during pregnancy is a major public health concern. The combined effects of cocaine and ethanol may be more injurious to the fetal nervous system than either drug alone. In order to identify areas of the brain vulnerable to concurrent exposure, we examined the expression of the immediate-early gene (IEG), c-fos, and cleaved caspase-3, the 'executioner' caspase in apoptosis. Pregnant rats were treated with either ethanol diet, cocaine binge, or both. At birth, the brains of fetuses exposed to cocaine exhibited an increase in Fos immunoreactivity in many brain regions. Prenatal exposure to ethanol did not increase Fos expression above that observed in control rats at early points after birth. However, Fos expression at 24 h after birth was higher after ethanol diet treatment in several brain regions, such as the amygdala, ventromedial hypothalamus, and medial thalamus. Only in the striatum did the combination of ethanol and cocaine cause greater Fos expression than either prenatal cocaine or ethanol alone. Increased cleaved caspase-3 expression was observed at the 24-h time point for both ethanol- and cocaine-exposed brains, most notably in the septum, retrosplenial cortex, and the hippocampus. Concurrent ethanol and cocaine exposure did not elevate cleaved caspase-3 expression beyond that of either drug alone. Analysis of the extent of c-fos and caspase-3 induction did not indicate a consistent relationship of expression in any of the drug treatment groups nor in any brain region. These results indicate that both prenatal cocaine and prenatal ethanol exposure increase Fos and cleaved caspase-3 expression in the developing brain in a time- and region-dependent manner, but that the combination of low dose, chronic ethanol, and binge cocaine does not cause greater apoptosis. PMID- 14741757 TI - Altered neuron-glia interactions in a low, chronic prenatal ethanol exposure. AB - Serotoninergic neurons, astrocytes and nitrergic system play an important role in central nervous system (CNS) development. These systems are altered in prenatal ethanol exposure (PEE) but ethanol (EtOH) effects may be very diverse under different conditions. In this study, we analyzed morphologically two serotoninergic mesencephalic nuclei and three prosencephalic areas of serotoninergic innervation in a model of pre- and postnatal low-ethanol exposure. Female Wistar rats were orally exposed to EtOH 6.6% (v/v), ad libitum, for 6 weeks before mating and during gestation and lactation while control group received water ad libitum. Twenty-day-old offspring (P21) brains were processed and immunoreactivity (IR) using antibodies against tryptophan hydroxylase (TPH), 5-HT, 5-HT transporter (5HTT), glial fibrillary acidic protein (GFAP), S-100B protein, 200-kDa neurofilaments (Nf-200) and neuronal nitric oxide synthase (nNOS) was evaluated. Dorsal and median raphe nucleus (DRN and MRN), hippocampus (Hipp), striatum (Strt) and frontal cortex (FCx) were studied by computer assisted image analysis. Relative optical density (ROD) of TPH-IR, 5-HT-IR and nNOS-IR neurons; cell area of GFAP-IR astrocytes; relative area of 5HTT-IR fibers and Nf-200-IR were evaluated. TPH-IR was increased in DRN and MRN and 5-HT-IR was increased only in MRN. 5-HTT-IR fibers and ROD of S-100B-IR astrocytes were increased in the three prosencephalic areas while GFAP-IR astrocytes were hypertrophied only in Hipp and FCx. Nf-200 expression was increased in Hipp and Strt and morphologically altered in the FCx. ROD of nNOS-IR neurons was increased in Strt and FCx but was not detected in Hipp. We have also detected morphological changes resembling accelerated development and maturation, and early aging. Considering the evidences of a close 5-HT-astroglial-NO relationship during CNS development the differential response of the studied regions is an interesting result that could be due to different gradients of development in the studied areas and/or different responses of those areas to the effects of a low pre- and postnatal ethanol exposure. PMID- 14741758 TI - Ethanol-induced neuronal death in organotypic cultures of rat cerebral cortex. AB - Ethanol can affect normal development of the cerebral cortex, e.g., it can disrupt cell migration and exacerbate cell death. In vitro studies using primary cultures or cell lines provide further evidence that cell migration and death are altered by ethanol exposure. Organotypic cultures are more complex than primary cell cultures, and maintain some normal connectivity, thus providing a "more in vivo-like" model of brain development. We predict that exposing organotypic cultures of fetal rat cerebral cortex to ethanol results in changes similar to those described in vivo. Organotypic cultures of brains from 16-day-old fetuses were exposed to ethanol (0, 200, 400 or 800 mg/dl) for 72 h. Stereological methods were used to assess the frequency of viable and dying cells. Dying cells were identified as having DNA with polyadenylated tails or as having condensed chromatin. A small amount of cell death was evident in the marginal zone (MZ) and cortical plate (CP) of control cultures. The MZ, normally a cell body-poor layer, was enriched with somata following exposure to 400 mg/dl ethanol. Ethanol-induced cell death in the MZ; the amount of cell death was doubled following exposure to 800 mg/dl ethanol. The CP was more sensitive than the MZ; cell death increased following treatment with 400 mg/dl ethanol. Thus, organotypic cultures show that ethanol disrupts neuronal migration and increases cell death in the developing cerebral cortex. The effects of ethanol were site-specific and concentration dependent. These changes are similar to those described in vivo. PMID- 14741759 TI - Serotonin fibre densities in subcortical areas: differential effects of isolated rearing and methamphetamine. AB - Serotoninergic neurons interact with dopaminergic cells on all levels and are physiologically affected by both isolated rearing (IR) and a single early methamphetamine (MA) injection. We therefore checked for anatomical effects of both interventions by immunohistochemically staining serotonin fibres and assessing fibre densities in the caudate-putamen (CPu), nucleus accumbens (NAc) and amygdala of Mongolian gerbils. IR led to significantly increased 5-HT fibre densities in the dorsal part of the CPu and in the central and basolateral amygdala. No effects were seen in the ventral CPu, in the NAc and in the lateral amygdala. The early MA injection resulted in a denser 5-HT innervation in the dorsomedial and ventromedial CPu, in the NAc shell of animals reared in an enriched environment and in the NAc core of both rearing conditions, leaving the lateral CPu and the amygdala unaffected. Thus, the single pharmacological versus the environmental challenge exerts an almost complementary effect on the 5-HT innervation in different areas of the brain, which demonstrates that systemic interactions, e.g. with dopaminergic and glutamatergic afferents, must be taken into account when the seemingly uniform 5-HT projections are investigated. PMID- 14741760 TI - Methamphetamine-induced gene expression profiles in the striatum of male rat pups exposed to the drug in utero. AB - Methamphetamine is a neurotoxic pychostimulant which affects monoaminergic and non-monoaminergic systems in the brain. Clinical studies in humans have found that exposure to methamphetamine in the developing embryo can cause significant behavioral and cognitive anomalies later in life. Exposure of animals to methamphetamine (METH) in utero can cause neurobehavioral effects that do not become apparent until young adulthood. In the present study, we sought to determine the effects of in utero METH exposure on the striata of perinatal rat pups using a recently developed 17 k cDNA microarray. We found that METH administration caused alterations in 913 genes according to strict criteria. These alterations include changes in genes that participate in signal transduction, heat shock responses and neuronal development. The majority of the changes in gene expression were more prominent at the 7-day time point. These observations suggest that in utero METH exposure might initiate molecular programs that significantly impact gene expression during the developmental period long after the last exposure to this drug. Thus, during development, METH exposure in utero might cause significant long-term changes in gene expression that might constitute, in part, some of the substrates for the behavioral and cognitive anomalies reported in the literature. PMID- 14741761 TI - Long-term effects of neonatal methamphetamine exposure in rats on spatial learning in the Barnes maze and on cliff avoidance, corticosterone release, and neurotoxicity in adulthood. AB - Methamphetamine (MA) is a commonly abused stimulant and because of its addictive properties, abusers may not cease use during pregnancy, thereby exposing the fetus to the drug. The consequences of such exposure remain largely unknown however data from animal models show that long-term deficits in spatial learning and memory in the Morris water maze (MWM) occur. In this study we explored the spatial learning ability of rats treated four times daily with MA (5 mg/kg/dose) during the sensitive period for induction of MWM deficits, postnatal days (P) 11 20, using a different maze. In adulthood the animals were tested in a non swimming spatial task, the Barnes maze, using either aversive (bright light) or appetitive (food reward) motivation. Approximately 30 days after behavioral testing, the pituitary and adrenal response to forced swim was assessed and susceptibility to MA-induced neurotoxicity measured. MA-treated animals tested in the aversive, but not the appetitive, version of the Barnes maze demonstrated spatial learning deficits. An attenuated corticosterone response in MA-treated animals was observed following forced swimming, however no differences in ACTH were found. Following acute MA administration in adulthood to all animals, the neonatally MA-treated animals displayed longer latencies to fall from a cliff than neonatally saline-treated rats given the same acute MA dose. This effect supports previous data showing hypoactivity in neonatally MA-treated animals. Acute MA treatment caused comparable striatal monoamine depletions in all groups, although females treated with MA as neonates displayed increased basal levels of corticosterone three days after the acute dose. These data demonstrate that MA administration during the neonatal period impairs spatial learning in an aversive non-swimming task and alters the adrenal response to a forced swim stressor, suggesting that the adrenal output during learning may contribute to the spatial learning deficits. PMID- 14741762 TI - Neonatal 3,4-methylenedioxymethamphetamine (ecstasy) alters dopamine and serotonin neurochemistry and increases brain-derived neurotrophic factor in the forebrain and brainstem of the rat. AB - Growing concerns surround the risk of fetal exposure to 3,4 methylenedioxymethamphetamine (MDMA; ecstasy). Prior animal studies using neonatal rats administered MDMA from postnatal days (P) 11-20 (a period approximating third trimester brain development in humans) have demonstrated long lasting decrements in serotonin (5-HT) and learning; however, no studies have examined the acute post-MDMA response of the brain at this early age. Specifically, it is of interest whether MDMA administration to neonatal rats produces the expected depletion of monoamines and whether the brain exhibits any ameliorative response to the pharmacologic insult. In the current study, this model was employed to determine whether forebrain and brainstem dopamine (DA) and 5-HT neurochemistry were altered 24 h after the last injection (P21), and whether brain-derived neurotrophic factor (BDNF) was upregulated in response to MDMA exposure. All forebrain structures examined (frontal cortex, hippocampus, and striatum) showed significant MDMA-induced reductions in 5-HT and its metabolite, 5-HIAA, and significant increases in the DA metabolite, HVA, as well as DA turnover (HVA/DA). In the brainstem, there were significant increases in 5-HIAA, HVA and DA turnover. BDNF was significantly increased (19-38%) in all forebrain structures and in the brainstem in MDMA-exposed neonates versus saline controls. These data suggest that MDMA exposure to the developing rat brain from P11-20 produces similar alterations in serotonin and dopamine neurochemistry to those observed from adult administrations. In addition, a compensatory increase in BDNF was observed and may be the brains ameliorative response to minimize MDMA effects. This is the first report demonstrating that MDMA exposure results in increased levels of BDNF and that such increases are correlated with changes in monoamine levels. Future research is needed to elucidate any deleterious effects MDMA-induced increases in trophic activity might have on the developing brain and to examine earlier gestational exposure periods in order to assess the risk throughout pregnancy. PMID- 14741763 TI - Nicotine is a developmental neurotoxicant and neuroprotectant: stage-selective inhibition of DNA synthesis coincident with shielding from effects of chlorpyrifos. AB - Although nicotine is now well recognized as a developmental neurotoxicant, it also may have neuroprotectant properties. In the current study, we used PC12 cells to characterize the specific developmental phases in which these effects are expressed. In undifferentiated cells, nicotine had a modest effect on DNA synthesis (10% reduction), which was nevertheless selective, as no significant reductions were seen for RNA or protein synthesis. The effects were blocked by mecamylamine, indicating mediation by nicotinic acetylcholine receptors. Initiation of differentiation with nerve growth factor, which greatly increases the receptor concentration, produced a commensurate increase in the sensitivity of DNA synthesis to nicotine, while RNA and protein synthesis again remained unaffected. The organophosphate insecticide, chlorpyrifos, also interferes with DNA synthesis in undifferentiated PC12 cells, but by mechanisms independent of nicotinic receptors. Accordingly, the effects of a combination of nicotine and chlorpyrifos should be additive. However, simultaneous exposure of undifferentiated cells to both agents produced less-than-additive effects at low concentrations of chlorpyrifos, and at high chlorpyrifos concentrations, nicotine produced outright protection: the combination of nicotine and chlorpyrifos had lesser effects than chlorpyrifos alone. The same neuroprotection was seen when cells were exposed to nicotine for 24 h, washed free of the drug for 24 h, and then exposed to chlorpyrifos. The results indicate that nicotine interferes with neural cell replication, with peak effects in early stages of differentiation. At the same time, nicotine promotes trophic actions that protect against neurotoxicants that work through other mechanisms. PMID- 14741764 TI - Method for producing coded micro-carrier and test method by using a novel type biochip. AB - This paper provides a method for producing a novel type coded micro-carrier. A simple and cost effective solution for bio-molecule applications was developed. Application relevant items such as manufacture process, biospecific interaction, and analysis method are discussed. For low cost fabrication, the use of LIGA-like process is suggested. LIGA-like process is used as a dry patterning process in which an intense beam of light from an excimer laser is used to pattern a material directly. This process has found extensive application in the microelectronics industry for patterning of polymer materials. The use of LIGA like techniques offers two attractive features: first, we can cut the polymer into many tiny micro-carriers with micrometer precision. Second, LIGA-like process allows to encode with high precision spatial information onto the micro carrier that can be used in the identification of the bio-molecule. This paper gives a description of the basic idea, describes the fabrication of the novel micro-carrier that we called "coded micro-carrier," and of the image processing algorithms used for the analysis of bio-molecules. This study also provides a test method for identifying a bio-molecule, which includes mixing several coded micro-carriers with the hybridized unknown bio-molecules; and identifying the codes on the micro-carrier via image recognition system. The numbers and types of the known micro-carrier can be flexibly adjusted according to the number of tested bio-molecules. PMID- 14741765 TI - Activity regulation of tyrosinase by using photoisomerizable inhibitors. AB - Enzymatic activity of tyrosinase was controlled on the basis of cis-trans photoisomerization of inhibitors, 4-azobenzene carboxylic acid (ACA) and 4,4' azobenzene dicarboxylic acid (ADCA). In the case of ACA, the cis form inhibited tyrosinase-catalyzed oxidation of L-tyrosine more strongly than the trans form. On the contrary, in the case of ADCA, the cis form was less inhibitory. The oxidation rate was controlled reversibly by light irradiation in the course of the reaction. In the presence of ACA, UV light irradiation, which isomerized trans to cis form, decelerated the tyrosine oxidation, while visible light irradiation, which isomerized backward, accelerated the reaction. In contrast, in the presence of ADCA, UV light accelerated and visible light decelerated the reaction. PMID- 14741766 TI - An improved beta-galactosidase reporter gene. AB - The coding sequence for the E. coli beta-galactosidase gene was codon-optimised for expression in mammalian cells. When expressed in mammalian cells the codon optimised gene results in the expression of beta-galactosidase at levels 15-fold higher than those resulting from an analogous construct containing the native E. coli gene sequence. RNA analysis suggests the enhancement of beta-galactosidase expression is due both to enhanced transcript stability and increased translational efficiency. When used in a lentiviral construct the codon-optimised gene results in an approximately five-fold increase in apparent titre, as determined by 5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside staining, in comparison to an analogous construct containing the native E. coli gene. Southern blot analysis shows this is due to an increased efficiency of detection of transduced cells. In addition, codon-optimisation results in the elimination of several cryptic splice acceptor sites that are present in the native E. coli gene sequence. In a lentiviral vector containing a 5' splice donor the use of the codon-optimised gene in place of the native E. coli beta-galactosidase gene resulted in increased amounts of un-spliced, full-length genomic RNA. Therefore, as a marker/reporter gene in mammalian cells the codon-optimised beta galactosidase gene has a number of advantages over the native E. coli gene sequence. A variant of the codon-optimised beta-galactosidase gene sequence that includes an effective nuclear localisation signal was also made. PMID- 14741767 TI - Optimised fermentation strategy for 13C/15N recombinant protein labelling in Escherichia coli for NMR-structure analysis. AB - A widely applicable cultivation strategy, which reduces the costs of expensive isotopes, is designed for maximal (98-100%) incorporation of [13C] and [15N] into labelled recombinant protein expressed in Escherichia coli, allowing better assignment of the resonances for NMR studies. Isotope labelling of the culture was performed throughout the complete process, starting from preculture. Sufficient biomass is first generated in a batch phase. Upon consumption of glucose, identified by a sharp drop of on-line monitored oxygen consumption, expression is induced and cultivation is continued under glucose-limited conditions as fed-batch process. Thereby a quantitative utilisation of the most expensive component [13C]-glucose is achieved, while the approximate amount of the [15N]-ammonium chloride to be incorporated is calculated from the scheduled biomass. The usefulness of the strategy is demonstrated with production of uniformly [13C/15N]-labelled tryparedoxin of Crithidia fasciculata. Ideal isotope incorporation and product quality is documented by MALDI-TOF mass spectrometry and two- and three-dimensional NMR spectra. PMID- 14741768 TI - Substrate specificity of native and mutated cytochrome P450 (CYP102A3) from Bacillus subtilis. AB - Within the Bacillus subtilis genome sequencing project, two monooxygenases (CYP102A2 and CYP102A3) were discovered which revealed a similarity of 76% to the well-known cytochrome P450 BM-3 (CYP102A1) of Bacillus megaterium. All enzymes are natural fusion proteins consisting of a heme domain and a reductase domain. We here report the cloning, expression and characterization of B. subtilis enzyme CYP102A3. The substrate specificity of this enzyme is similar to that of B. megaterium CYP102A1, which hydroxylates medium-chain fatty acids in subterminal positions. A double mutant was prepared that hydroxylates a number of other substrates, which do not bear any resemblance to the natural substrate of this enzyme family. PMID- 14741769 TI - Characterisation of steryl esterase activities in commercial lipase preparations. AB - Triglycerides, steryl esters, resin acids, free fatty acids and sterols are lipophilic extractives of wood (commonly referred to as pitch or wood resin) and have a negative impact on paper machine runnability and quality of paper. Thus, enzymes capable of modifying these compounds would be potential tools for reducing pitch problems during paper manufacture. In this work, 19 commercial lipase preparations were tested for their ability to degrade steryl esters, which may play a significant role in the formation and stabilisation of pitch particles. Six lipase preparations were shown to be able to degrade steryl esters. Lipase preparations of Pseudomonas sp., Chromobacterium viscosum and Candida rugosa were shown to have the highest steryl esterase activities. The enzymes were able to hydrolyse steryl esters totally in the presence of a surfactant (Thesit). Up to 80% of the steryl esters were degraded in aqueous dispersion. Preliminary characterisation of the enzymatic activities revealed that the lipase preparation of Pseudomonas sp. could be the most potential enzyme in industrial applications. The steryl esterase activity of this preparation was stable over a broad pH range and the enzyme was able to act efficiently at pH 6 10 and at temperatures up to 70 degrees C. PMID- 14741770 TI - Intelligent real-time performance monitoring and quality prediction for batch/fed batch cultivations. AB - Supervision of batch bioprocess operations in real-time during the progress of a batch run offers many advantages over end-of-batch quality control. Multivariate statistical techniques such as multiway partial least squares (MPLS) provide an efficient modeling and supervision framework. A new type of MPLS modeling technique that is especially suitable for online real-time process monitoring and the multivariate monitoring charts are presented. This online process monitoring technique is also extended to include predictions of end-of-batch quality measurements during the progress of a batch run. Process monitoring, quality estimation and fault diagnosis activities are automated and supervised by embedding them into a real-time knowledge-based system (RTKBS). Interpretation of multivariate charts is also automated through a generic rule-base for efficient alarm handling. The integrated RTKBS and the implementation of MPLS-based process monitoring and quality control are illustrated using a fed-batch penicillin production benchmark process simulator. PMID- 14741771 TI - Enzymatic versus chemical deinking of non-impact ink printed paper. AB - Enzymatic versus chemical deinking is examined for MOW and photocopy prints. Several enzymatic preparations and two fibre/ink particle separation methods are tested. Deinking was monitored by image analysis and standard pulp and paper characterisation procedures. The effectiveness of the fibre/ink particle separation method depends on the ink particle's size: for smaller particles a washing step is recommended whereas for larger particles, the use of flotation is necessary. The enzymatic treatment is a competitive alternative for MOW and photocopy paper deinking. However, the process requires the selection of an adequate enzymatic preparation for each paper grade. PMID- 14741772 TI - Chance or necessity? Insertional mutagenesis in gene therapy and its consequences. AB - Recently, unusual forms of leukemias have developed as complications following retroviral transfer of potentially therapeutic genes into hematopoietic cells. A crucial component in the pathogenesis of these complications was the upregulation of a cellular proto-oncogene by random insertion of the retroviral gene transfer vector. These findings have great implications for the genetic manipulation of somatic stem cells in medicine. This review discusses the extent to which the random oncogene activation may have required disease-specific stimuli of the transgene and the hematopoietic milieu to become leukemogenic. Based on these considerations, we propose approaches to risk prediction and prevention. PMID- 14741773 TI - Myoblast transplantation for cardiac repair: a clinical perspective. AB - The incidence of heart failure is achieving epidemic proportions. Adult human myocytes cannot regenerate because these cells do not re-enter the cell cycle. In patients with heart failure, myoblast transplantation is emerging as a potential therapeutic option to augment the function of remaining myocytes. Both skeletal myoblasts and autologous bone marrow cell transplantation, after intensive preclinical experimental animal studies, have entered phase I safety studies in humans. Most of these clinical trials have involved small groups of patients and cell transplantation was carried out as an adjunct to coronary revascularization. Preliminary results show that the procedure is safe and leads to improved myocardial function. This paper reviews and summarizes the outcome of these phase I trials involving skeletal myoblast transplantation. PMID- 14741774 TI - Reversal of ongoing thermal hyperalgesia in mice by a recombinant herpesvirus that encodes human preproenkephalin. AB - Herpesvirus-mediated transfer of the human preproenkephalin gene to primary afferent nociceptors prevents phasic thermal allodynia/hyperalgesia in mice. It is not known, however, whether similar viral treatments would reverse ongoing or chronic pain and allodynia/hyperalgesia. To this end, mice were given intrathecal injections of pertussis toxin (PTX), which produces a weeks-long thermal hyperalgesia apparently by uncoupling certain G proteins from inhibitory neurotransmitter receptors. This treatment produced profound thermal hyperalgesia in both Adelta and C-fiber thermonociceptive tests lasting at least 6 weeks. However, treatment of skin surfaces with an enkephalin-encoding herpesvirus, but not control virus or vehicle, completely reversed this hyperalgesia. This profound anti-hyperalgesia was observed for both Adelta- and C-fiber-mediated responses. Interestingly, however, while the anti-hyperalgesic effect of the enkephalin-encoding virus on C-fiber-mediated responses was reversed by intrathecal application of micro or delta opioid antagonists, only delta antagonists reversed the effect of this virus on Adelta hyperalgesia. Thus, virus mediated delivery of the proenkephalin cDNA reverses thermal hyperalgesia produced by PTX-induced ribosylation of inhibitory G proteins by an opioid mediated mechanism. These results suggest that herpesvirus vectors encoding analgesic peptides may be useful in attenuating centrally mediated, ongoing neuropathic pain and/or hyperalgesia. PMID- 14741775 TI - Improved effects of viral gene delivery of human uPA plus biliodigestive anastomosis induce recovery from experimental biliary cirrhosis. AB - Gene therapy may represent a new avenue for the development of multimodal treatment for diverse forms of cirrhosis. This study explores the potential benefits of combining adenovirus-mediated human urokinase-plasminogen activator (AdHuPA) gene delivery and biliodigestive anastomosis to enhance the therapeutic efficacy of each treatment alone for cholestatic disorders resulting in secondary biliary cirrhosis. In an experimental model of secondary biliary cirrhosis, application of 6 x 10(11) vp/kg AdHuPA adenovirus vector resulted in 25.8% liver fibrosis reduction and some improvement in liver histology. The relief of bile cholestasis by a surgical procedure (biliodigestive anastomosis) combined with AdHuPA hepatic gene delivery rendered a synergistic effect, with a substantial 56.9 to 42.9% fibrosis decrease. AdHuPA transduction resulted in clear-cut expression of human uPA protein detected by immunohistochemistry and induction of up-regulation in the expression of metalloproteinases MMP-3, MMP-9, and MMP-2. Importantly, functional hepatic tests, specifically direct bilirubin, were improved. Also, hepatic cell regeneration, rearrangement of hepatic architecture, ascites, and gastric varices improved in cirrhotic rats treated with AdHuPA but not in counterpart AdGFP cirrhotic animals. We believe this might represent a novel therapeutic strategy for human cholestatic diseases. PMID- 14741776 TI - Gene therapy for liver transplantation using adenoviral vectors: CD40-CD154 blockade by gene transfer of CD40Ig protects rat livers from cold ischemia and reperfusion injury. AB - Liver injury induced by ischemia/reperfusion (I/R) is the prime factor in delayed or loss graft function following transplantation. CD4+ T lymphocytes are key cellular mediators of antigen-independent inflammatory response triggered by I/R. We attempted to modulate rat liver I/R injury by targeted gene therapy with CD40Ig, which blocks the CD40-CD154 costimulation pathway. One hundred percent of Ad-CD40Ig-pretreated orthotopic liver transplants (OLTs) subjected to 24 h of cold (4 degrees C) ischemia survived > 14 days (vs 50% in untreated/Ad-beta-gal groups). Ad-CD40Ig treatment decreased sGOT levels and depressed neutrophil infiltration, compared with controls. These functional data correlated with histological Suzuki's grading of hepatic injury, which in untreated/Ad-beta-gal groups showed severe necrosis (> 60%) and moderate to severe sinusoidal congestion; the Ad-CD40Ig-pretreated group revealed minimal sinusoidal congestion/necrosis. Unlike in controls, OLT expression of mRNA coding for IL 2/IFN-gamma remained depressed, whereas that of IL-4/IL-13 reciprocally increased in the Ad-CD40Ig group. Ad-CD40Ig reduced frequency of TUNEL+ cells and pro apoptotic Caspase-3, but enhanced antioxidant HO-1 and anti-apoptotic Bcl-2/Bcl xl expression. Thus, prolonged blockade of CD40-CD154 by CD40Ig exerts potent cytoprotection against hepatic I/R injury. These results provide the rationale for a novel gene therapy approach to maximize the organ donor pool through the safer use of liver transplants exposed to prolonged cold ischemia. PMID- 14741777 TI - Insulin-like growth factor I plasmid therapy promotes in vivo angiogenesis. AB - Angiogenesis, the formation of neovessels from the endothelium of preexisting vessels, is stimulated by soluble angiogenic factors. Insulin-like growth factor I (IGF-I) stimulates myogenesis and induces nerve regeneration after injury, and it has been shown to stimulate angiogenesis. However, the in vivo angiogenic effects of IGF-I in regenerating and diabetic muscle have yet to be described. Therefore, we studied the effects of human IGF-I (hIGF-I) delivered by a plasmid mediated therapy on angiogenesis in mouse models of these two conditions. Plasmid hIGF-I was delivered to the injured tibialis muscle by direct intramuscular injection followed by electroporation. Initial experiments compared two muscle specific hIGF-I-expressing constructs containing either a skeletal actin 3'UTR (pAV2001) or a human growth hormone (GH) 3'UTR (pAV2002). Skeletal actin 3'UTR mediates sequestration of hIGF-I in the muscle and was more active, while the GH 3'UTR mediated release of IGF-I into the circulation. Treatment of regenerating muscle with pAV2001 and sequestration of IGF-I in muscle led to increased expression of vascular endothelial growth factor (VEGF) and VEGF receptors fetal liver kinase-1 and FmS-like tyrosine kinase receptor-1, as well as platelet endothelial cell adhesion molecule-1, on endothelial cells. These results indicate that IGF-I can amplify angiogenic responses in regenerating muscle. In a mouse diabetic model, plasmid-mediated IGF-I therapy reversed diabetic microangiopathy, as shown by increased angiogenesis and arterial flow as analyzed by Doppler imaging. These studies show that plasmid IGF-I delivery and sequestration in muscle can augment angiogenesis in regenerating muscle and increase blood flow and angiogenesis in the diabetic limb. PMID- 14741778 TI - Anti-tumor efficacy of human angiostatin using liver-mediated adeno-associated virus gene therapy. AB - Angiostatin is a potent endogenous inhibitor of angiogenesis and tumor growth in vivo. The therapeutic potential of adeno-associated viral (AAV) gene delivery of angiostatin in modulating tumor growth in vivo was evaluated. Sustained levels of angiostatin were detected in the sera of mice for up to 6 months after they received a single injection of AAV-angiostatin. AAV-mediated stable expression of angiostatin inhibited tumor burden in the highly aggressive B16F10 melanoma and Lewis lung carcinoma (LLC) models of experimental metastasis. Moreover, AAV angiostatin prolonged survival in B16F10 and LLC tumor-bearing mice compared to control groups. Anti-tumor efficacy was consistently observed when angiostatin serum levels of 15-50 ng/ml were detected following gene transfer, but the effect was minimal when the levels were lower or higher than this range. The combination of AAV-angiostatin gene therapy with chemotherapy was also shown to extend marginally the survival of mice bearing preestablished human tumors; however, the effect was evident only within a narrow dose of circulating angiostatin. These studies demonstrate the feasibility of using AAV anti-angiogenic gene therapy as a cancer treatment modality and suggest that the optimal anti-tumor efficacy of angiostatin following gene transfer may be limited to a narrow dose range. PMID- 14741780 TI - Ligand-modified vesicular stomatitis virus glycoprotein displays a temperature sensitive intracellular trafficking and virus assembly phenotype. AB - The production of potentially targetable VSV-G-pseudotyped retrovirus vectors has been hampered by inadequate understanding of the structure-function relationships of the VSV-G protein. In these studies we demonstrate assembly and production of MLV-based and HIV-1-based vector particles using VSV-G proteins modified by the insertion of a peptide ligand into a site corresponding to amino acid position 24 of the native VSV-G molecule. The inserted ligand represents the decapeptide encoding the collagen-binding domain of von Willebrand factor. We have used deconvolution microscopy to demonstrate that the modified VSV-G molecules sequester in perinuclear structures and are unavailable for assembly of infectious virus particles at the cell surface under standard tissue culture conditions at 37 degrees C. In contrast, at a lower permissive temperature of 30 degrees C, the modified VSV-G protein traffics appropriately to the cell surface and participates in useful titers. Furthermore, VSV-G-pseudotyped MLV-based and HIV-1-based vectors displaying the collagen-binding domain demonstrate a statistically significant increased attachment to a collagen matrix as indicated by an ELISA-like cell binding assay and by a focus transduction assay. PMID- 14741779 TI - Adenovirus-mediated transfer of a minigene expressing multiple isoforms of VEGF is more effective at inducing angiogenesis than comparable vectors expressing individual VEGF cDNAs. AB - To assess the hypothesis that angiogenic gene therapy with the genomic form of vascular endothelial growth factor (VEGF) expressing the three major isoforms could be more potent than a vector expressing a single isoform, we designed an adenovirus vector (AdVEGF-All) expressing a VEGF cDNA/genomic hybrid gene. AdVEGF All expressed all three major isoforms (121, 165, 189) in a 2:2:1 ratio. AdVEGF All was 100-fold more potent than cDNA vectors expressing VEGF 121, 165, or 189 in restoring blood flow to the ischemic mouse hind limb. Interestingly, a mixture of Ad vectors individually expressing the VEGF 121, 165, and 189 cDNAs was equipotent to an equivalent dose of AdVEGF-All. Thus, a mixture of VEGF isoforms provides a more potent angiogenic response than a single isoform, suggesting that the individual isoforms function synergistically, an observation with important implications for gene and recombinant protein therapy. PMID- 14741781 TI - Efficient retroviral vector targeting of carcinoembryonic antigen-positive tumors. AB - Many gene therapy approaches require specific, efficient gene delivery to cells in vivo. To target colorectal tumors we fused a single-chain variable fragment (scFv) directed against carcinoembryonic antigen (CEA) to the amphotropic murine leukemia virus envelope. A proline-rich hinge and matrix metalloprotease (MMP) cleavage site linked the two proteins. Following attachment to CEA, MMP cleavage of the envelope at the cell surface removed the scFv and proline-rich hinge, allowing transduction. This allowed selective targeting of CEA-positive cells in vivo after injection of producer cells at the site of the tumor, with up to 10% of cells within a CEA-positive tumor xenograft becoming transduced. Intraperitoneal injection of amphotropic producer cells resulted in transduction of cells in spleen, liver, and kidney, which was not detected when CEA-targeted producer cells were used. These results demonstrate the feasibility of using targeted retroviral vectors for in vivo gene delivery to tumors. Furthermore, the lack of transduction of host cells eliminates the risk of insertional mutagenesis leading to transformation of host hematopoietic cells. PMID- 14741782 TI - Lentivectors for regulated and reversible cutaneous gene delivery. AB - Systemic administration of therapeutic proteins has value in treating a wide variety of disorders, including erythropoietin (Epo)-responsive anemias. Recombinant proteins, however, are costly and require repeated injections, while gene delivery approaches have suffered from inefficiency and difficulties with regulation. The skin effectively delivers polypeptides to the circulation, and improved approaches would support sustainable, topically regulated protein expression after a single vector injection. Toward this goal, we generated lentivectors in which both gene delivery and persistence in skin are regulated by administration of distinct steroid ligands. Following a single injection of regulated lentivector into human skin regenerated on immunodeficient mice, topical glucocorticoid ligands regulated Epo levels and hematocrit over time. Abrogation of gene delivery was achieved by both glucocorticoid cessation and proviral excision via a 4-hydroxytamoxifen-inducible Cre recombinase. These findings establish an approach to durable, topically controlled systemic delivery of therapeutic proteins from human skin tissue. PMID- 14741783 TI - Transduction patterns of pseudotyped lentiviral vectors in the nervous system. AB - We have developed a non-primate-based lentiviral vector based on the equine infectious anemia virus (EIAV) for efficient gene transfer to the central and peripheral nervous systems. Previously we have demonstrated that pseudotyping lentiviral vectors with the rabies virus glycoprotein confers retrograde axonal transport to these vectors. In the present study we have successfully produced high-titer EIAV vectors pseudotyped with envelope glycoproteins from Rhabdovirus vesicular stomatitis virus (VSV) serotypes (Indiana and Chandipura strains); rabies virus [various Evelyn-Rokitnicki-Abelseth ERA strains and challenge virus standard (CVS)]; Lyssavirus Mokola virus, a rabies-related virus; and Arenavirus lymphocytic choriomeningitis virus (LCMV). These vectors were delivered to the striatum or spinal cord of adult rats or muscle of neonatal mice by direct injection. We report that the lentiviral vectors pseudotyped with envelopes from the VSV Indiana strain, wild-type ERA, and CVS strains resulted in strong transduction in the striatum, while Mokola- and LCMV-pseudotyped vectors exhibited moderate and weak transduction, respectively. Furthermore ERA- and CVS pseudotyped lentiviral vectors demonstrated retrograde transport and expression in distal neurons after injection in brain, spinal cord, and muscle. The differences in transduction efficiencies and retrograde transport conferred by these envelope glycoproteins present novel opportunities in designing therapeutic strategies for different neurological diseases. PMID- 14741784 TI - Morphological analysis and lentiviral transduction of fetal monkey bone marrow derived mesenchymal stem cells. AB - We explored the transduction kinetics of HIV-1-derived lentiviral vectors containing the CMV, EF1alpha, or PGK promoter expressing EGFP in fetal rhesus monkey bone marrow-derived mesenchymal stem cells (rhMSC). Studies included the effects of transduction (MOI 0-100) on growth, cell cycle, and differentiation toward an osteogenic lineage. Flow cytometric analysis indicated an approximate 8 to 10-fold greater quantity of EGFP-expressing rhMSC when cells were transduced with the CMV or EF1alpha promoter compared to PGK, although quantitative PCR revealed no differences at the DNA level. The CMV promoter initially expressed 10 to 100-fold higher levels of EGFP compared to EF1alpha or PGK, respectively, at increasing MOI, although a significant decline in transgene expression was observed posttransduction and with advancing passage (P < 0.01), whereas a significant increase in the level of expression was observed over time with the EF1alpha promoter. At an MOI of 100, a transient arrest at the S phase of the cell cycle was observed for both vector constructs. Transduced rhMSC differentiated toward an osteogenic lineage comparable to untransduced rhMSC and showed equivalent levels of alkaline phosphatase activity. These findings suggest that the SIN HIV-1-derived lentiviral vectors used in these studies can efficiently transduce rhMSC in vitro (CMV > EF1alpha > PGK) without inhibiting differentiation potential, although the cell cycle was transiently altered at high MOI PMID- 14741785 TI - Toward PKU enzyme replacement therapy: PEGylation with activity retention for three forms of recombinant phenylalanine hydroxylase. AB - Phenylketonuria (PKU) is a disease in which phenylalanine and phenylalanine derived metabolites build up to neurotoxic levels due to mutations in the phenylalanine hydroxylase gene (PAH). Enzyme replacement therapy is a viable option to supply active PAH. However, the inherent protease sensitivity and potential immunogenicity of PAH have precluded adoption of this approach. In this report, we have used polyethylene glycol derivatization (PEGylation) to produce protected forms of PAH for potential therapeutic use. Three recombinantly produced PAH enzymes were reacted with activated PEG species, with the aim of developing a stable and active PKU enzyme replacement. Tetrameric full-length human PAH, dimeric double-truncated (DeltaN102-DeltaC428) human PAH, and monomeric Chromobacterium violaceum PAH were PEGylated with succinimidyl succinate polyethylene glycol of molecular weight 5000 or 20,000 Da. Characterization of the PEGylated species was accomplished with MALDI-TOF mass spectrometry, SDS-PAGE, and specific activity measurements using ESI mass spectrometry. All PEG-derivatized PAH species retained catalytic activity, and, at low numbers of PEG molecules attached, these PEGylated PAH proteins were found to be more active and more stable than their non-derivatized PAH counterparts. PMID- 14741789 TI - The importance--and limits--of best evidence medical education. PMID- 14741786 TI - Delivery and protection of adenoviruses using biocompatible hydrogels for localized gene therapy. AB - Localized gene delivery for repair of bone defects requires appropriate carriers for the gene therapy vectors. The objective of this study was to determine if hydrogels can control temporal and spatial delivery of adenovirus for localized gene therapy. Adenovirus expressing beta-galactosidase was suspended in liquid or fibrin or collagen gels of varied concentrations and incubated prior to testing its bioactivity. The bioactivity of the virus was determined by exposing fibroblasts to the medium, the gels, or the elution medium from the gels. Bioactivity of adenovirus suspended in medium or collagen decreased to half maximal activity after 15 h of incubation. In contrast, virus suspended in fibrin exhibited a threefold extension of bioactivity and did not reach half-maximal activity for 45 h. Bioactivity of adenovirus in hydrogels was determined to be a function of the gel concentration. In vivo experiments involved intramuscular implantation of BMP-7-expressing adenovirus in collagen, fibrin, or liquid in nude mice for 1, 2, or 4 weeks. Bone formation was observed only after 4 weeks, with bone formation occurring in 80% of muscles implanted with fibrin or collagen and 50% of muscles implanted with liquid. Fibrin gel also led to significantly larger ossicles, indicating that fibrin may offer protection from loss of infectivity both in vivo and in vitro. These results demonstrated that hydrogels may be used as carriers to control delivery of the virus and resultant tissue regeneration. PMID- 14741790 TI - Medical education, research, and scientific thinking in the 21st century (part one of three). AB - BACKGROUND: During the 20th century medical education has been preoccupied largely with discussions of the venues and methods for teaching. Little attention has been paid to what should be learned about the scientific paradigm underlying research and practice. A 17th century model has gradually produced a technically efficient but increasingly narrow, monocausal, reductionistic view of health and disease. This "belief system" fails to accommodate or explain the meaning and impact on patients' health of diverse internal and external experiences and influences. During this period new physics and systemic views of biosystems have extended the Newtonian scientific paradigm beyond its materialistic boundaries, which still determines most of the medical sciences. METHODS: A broad range of historical and contemporary scientific literature is examined in support of four central questions addressed in this three-part series: Is there a reason to examine these matters now? How is medical scientific thinking influenced by the general reorientation of science during the 20th century? Is there is a reason to examine the impact of these changes on medicine now? Will a change of paradigm affect medical practice, research, and education? RESULTS: The extraordinarily productive contemporary biomedical model should be expanded to include meaningful information about how each patient's experiences impinge on health status. CONCLUSIONS: Family physicians, together with collaborators in the biological and behavioral sciences and other health professionals, should undertake rigorous research to establish the validity of the expanded paradigm espoused. Its impact could be profound on practice, research, education, and policies. PMID- 14741791 TI - The Quality of Australian Health Care Study: implications for education of failure in quality and safety of health care. AB - Medical and health professional education needs to renew regularly the perspective from which it defines curriculum. The theme of this paper is that the study of health service in action will identify educational priorities for any country with the particular aim of improving the quality of health care and the effectiveness of clinicians within the system of health care. The Quality of Australian Health Care Study is the largest study of its kind to date. It is a study of adverse outcomes in health care resulting from error. From a study of causes, circumstance and strategies for prevention an educational agenda emerges, dealing both with clinical aspects of care and the less well-documented role of system errors. Studies of system errors in industry and other high risk occupations and systems are now lending insight to issues of error and safety in health care. A new agenda for health professional curriculum is the study of health services in action-the anatomy, physiology and pathology of health systems. Eight educational priorities were recommended to the National Task Force to improve patient safety and quality of care. These are discussed together with a broad strategy for curriculum implementation. The need for a health system that supports and informs practitioners involved in clinical and system errors, the extension of training to incorporate health system managers, and the extension of the consideration of safety to public awareness are discussed. PMID- 14741792 TI - Health professions education for adapting to change and for participating in managing change. AB - This paper outlines a case for a new approach to the education of future professionals. The magnitude and potential seriousness of changes to be anticipated in the first half of the new century challenge institutions of higher education to prepare their students to become able to adapt themselves to change and to participate in the management of change--not only in relation to their own profession, but more widely for the benefit of society at large. The Network of Community-Oriented Educational Institutions for Health Sciences is planning three major programs which are designed to help meet this challenge in the 21st century. The first of these programs will set out to identify generic competencies that are needed for adapting to change and for participating in managing change. This is to include not only profession-specific changes, but also changes that affect the well-being of society as a whole. The second program will address the related need to develop educational interventions that are designed to foster competencies for interprofessional and intersectoral collaboration. The third of these programs accepts that serious attention to effective education for the professions will require institutionalization of recognition and reward of creativity and commitment in education. PMID- 14741793 TI - Examining and recording clinical performance: a critique and some recommendations. AB - Clinical performance is too complex and interactive for measurement. Judgment is always necessary for its assessment. Experienced clinicians judge trainee performance on many small details. This clinical judgment turns on the trainee's handling of important details in the patient and the malady. But the recording of performance retreats to categories and checklists that contain nothing of those critical details or the trainee's judgment. Checklists are incapable of identifying what actually happened, and 'could do' categories have no predictive accuracy in asserting what cases a trainee can actually manage. Clinical examinations have even been subverted by the naive, pseudorational error that competence is defined by obedience to doing exactly what someone else expects you to do in every case, as in an OSCE examination. Cases are the unit of clinical practice. The clinical curriculum should be comprised of the critical core cases the trainee must be able to handle in each discipline. Case management, procedural skills and professional behavior can be assessed accurately only in the context of daily clinical work. Formal examinations lack the range of cases and open-ended time that allow examiners to explore a trainee's case knowledge and judgment. Habitual behavior can be assessed only by observing habitual behavior in everyday practice. Assessment and recording should take place only in real world settings, focused on performance on the core cases trainees must be competent to manage. PMID- 14741794 TI - Introducing community-based teaching of third year medical students: outcomes of a pilot project one year later and implications for managing change. AB - INTRODUCTION: British undergraduate medical education is undergoing change, including a greater emphasis being placed on community-based teaching. These changes need to be evaluated for their educational outcomes, but there also needs to be a review of the process of introducing change and its subsequent management. THE SETTING: During the academic year 1996/97 a new project was piloted at Leeds University. Fifty-three third year medical students were attached in groups of four to general practitioner tutors in a primary care setting for four days in order to improve consultation skills. There was an emphasis on adopting a patient-centered approach, particularly asking patients about their concerns. METHOD: These students were asked to fill in a questionnaire to determine whether the community-based teaching has made any lasting impression on their attitudes and performance. The response rate was 80%. The students commented that even only four days of community-based teaching had helped them realize the importance of asking about patients' concerns. They also reported beginning to concentrate on psychosocial issues while talking to patients, issues that are often ignored by medical students. The students' comments are contrasted with those of the hospital-based tutors, some of whom have looked less than favorably on the project. DISCUSSION: The introduction of any new learning experience needs to be evaluated both in the short term and in the long term. This study begins to address long-term evaluation and suggests that a brief attachment can be memorable to students and beneficial one year later. PMID- 14741795 TI - Commentary on introducing Community-based teaching of third year medical students: outcomes of a pilot project one year later and implications for managing change. The critical role of faculty in supporting and sustaining innovation. PMID- 14741796 TI - The role of the community in educating medical students: initial impressions from a new program. AB - PURPOSE: The Keeping Families Healthy (KFH) program at the University of Florida College of Medicine is a new community-based course designed to improve physician patient relationships. This paper describes the experience of the first-year medical students' home visits, which were developed to foster health promotion and disease prevention. APPROACH: This two-semester course offers first-year medical students the opportunity to integrate prevention, service, and humanism into the established educational curriculum. During the course the learners have opportunities to interact with families who have volunteered to serve in partnership with the College of Medicine faculty as community lay teachers. The program provides a valuable service to the participating families by helping them identify useful community resources, and by formulating healthy care plans for prevention of illness and stabilization of chronic illness. CONCLUSIONS: Community-based learning and home visits expose students to personal travails (e.g. lack of financial resources) in a way that cannot be addressed in traditional settings. This exposure may involve students in community-building strategies that can assist not only individual patients, but also communities. Additionally, home visits provide an opportunity for pre-clinical students to have an active rather than passive role in their education. PMID- 14741797 TI - Case design to emphasize population health concepts in problem-based learning. AB - BACKGROUND: Medical training traditionally focuses on disease diagnosis and management. The need to incorporate preventive medicine, economics, and health promotion is increasingly apparent. Because problem-based learning (PBL) encourages multidisciplinary thinking, it is ideal for linking traditional medical education and population-oriented training. Although use of PBL has grown in medical education, cases typically focus upon patho-physiology, diagnosis, and therapy of individuals. Even when cases are intended to integrate multidisciplinary topics such as behavioral sciences or prevention, the biological aspects are emphasized. PURPOSE: To describe approaches to case design that emphasize population perspectives of health. DESCRIPTION: Specific examples drawn from actual cases we have used illustrate how five basic components of a case--namely, title, context, intrigue, indicators of problem resolution, and tight structure--facilitate discussion of, and enhance concern for, population issues. CONCLUSION: The literature indicates that health professional students tend to favor biological over population content in clinical cases. We illustrate how population content can be represented in specifically designed cases. PMID- 14741799 TI - Focus groups: a qualitative method complementing quantitative research for studying culturally diverse groups. AB - CONTEXT: Focus groups are becoming an important method for conducting qualitative research in health care. This strategy enables information to be gathered on the perceptions, beliefs, and values of a group's participants and is particularly well suited to addressing cultural characteristics that impact on a population's health status. As nations become more culturally diverse, qualitative research will likely play a growing role in helping health professions educators develop appropriate educational programs and in helping researchers better understand the needs of minorities and other vulnerable populations who are experiencing disparities in health care. OBJECTIVES: The purposes of this paper are to introduce the usefulness of a qualitative research strategy as an adjunct to quantitative survey research, and to describe briefly how researchers and educators at the Charles R. Drew University of Medicine and Science (Drew) have utilized this strategy when conducting combined qualitative and quantitative research. DISCUSSION: Focus group research has been successfully used to develop culturally adapted surveys, to develop educational programs, and to conduct needs assessments at Drew, which serves a culturally diverse urban population. PMID- 14741800 TI - Why German medical students abandon dissertations. AB - In Germany, conducting a research project followed by writing a thesis is necessary to receive the title "Doctor" but is not necessary to work as a physician. To determine the reasons that students abandon their dissertations, we conducted and analyzed a survey among 160 fifth and sixth-year medical students at the University of Wuerzburg. Fifteen of 160 respondents (9%) reported to have stopped working on their original project before starting their current replacement thesis research project. Among these 15, the major reasons for abandoning the original research project were: seven (47%) reported lack of supervision, three (20%) reported no recognizable progress, and another three (20%) reported an unacceptable burden of work. Unfamiliarity with experimental methods led two students (13%) to stop working on their original research project. Close supervision and a thorough training remain necessary to prevent students from abandoning medical dissertations and thus should be an essential part of every research project in which medical students are involved. PMID- 14741802 TI - An interview of Ronak Iqbal. Interview by Jane Westberg. PMID- 14741801 TI - An interview of Dr. John D. Hamilton. Interview by Jane Westberg. PMID- 14741803 TI - Qualitative research: a consumer's guide. AB - Qualitative research is best characterized as a family of approaches whose goal is understanding the lived experience of persons who share time, space and culture. Although they are often judged as a single entity, the approaches actually vary in their theoretical assumptions and canons of evidence. Four qualitative research domains that are currently used in studying education for health are reviewed here. They are ethnographic/field work approaches, use of interviews and surveys, audiovisual records, and the study of documents. Characteristics of each domain and brief examples are provided. In addition to introducing the four research domains, we offer some general guidelines on how to be a good consumer of qualitative research. We pose a series of questions about the importance of the research question, study design, and trustworthiness of qualitative research results. In addition, we focus on how research results are presented and discussed. We conclude with the observation that qualitative research approaches are only as good as the questions they set out to illuminate. In the arena of education for health a number of good and important questions remain unaddressed and would benefit by being studied using qualitative research approaches. PMID- 14741805 TI - Preventive care education by physicians: a call for action. PMID- 14741807 TI - Forces on medical education: reflections on the conference celebrating the 40th anniversary of the Department of Medical Education, University of Illinois Chicago. PMID- 14741815 TI - Non-ablative scar revision using a long pulsed frequency doubled Nd:YAG laser. AB - OBJECTIVE: Unsightly scars often are the only reminder of a previous surgical or traumatic wound. Surgical or ablative scar revision is sought by patients, sometimes unnecessarily. When the aesthetic drawback is mainly a result of hypervascularity or hyperpigmentation, these problems can be specifically targeted with a wavelength that is well absorbed by the two above mentioned chromophores. Some degree of epidermal tightening can also be achieved, which is sometimes useful in slightly atrophic scars. The average improvement after 2-3 sessions was 81% (75%-100%) clearance, as judged by an independent observer who reviewed pre- and post-treatment photographs. No undesired effects were reported. All our patients were satisfied and required no further treatment. METHODS: Selective photothermolysis by means of a long pulsed frequency doubled Nd:YAG laser (DioLite 532, IRIDEX Corporation, Mountain View, CA, USA) was used to eliminate the unsightly vascular and pigmented components of 23 mature scars (scars older than 2 years) in 22 consecutive patients. Energy densities of 17-22 j/cm2 were used with a 500 micron spot, or 65-90 j/cm2 with a 200 micron spot. RESULTS: overall scar clearance averaged 81% after 2.4 treatments. Facial scars showed the best clearance averaging 94% after 2 treatments. Inframammary scars were the most difficult to clear averaging 46%. Postoperative undesired effects were immediate erythema and swelling that subsided within 2-10 hours and microcrusting on 19/22 (88%) patients that resolved within one week. No other temporary or permanent undesired effects such as purpura, hypo- or hyperpigmentation were noticed, even in patients with darker skin types. PMID- 14741816 TI - Efficacy of long- and short pulse alexandrite lasers compared with an intense pulsed light source for epilation: a study on 532 sites in 389 patients. AB - BACKGROUND: Undesirable hair growth presents a significant problem for many patients, and photoepilation has become a very popular procedure in aesthetic and cosmetic practice. Among the systems used are the long- and short-pulsed alexandrite lasers (LP-Alex, SP-Alex) and intense pulsed light (IPL) sources. The present study retrospectively examined the outcome of these systems from the viewpoint of efficacy and side effects. PATIENTS AND METHODS: Three hundred and eighty-nine patients (370 females and 19 males, mean age 36.4 yrs, skin types II V) were admitted to the study, with a total of 532 treated sites. They were treated either with the LP-Alex, SP-Alex or IPL. Subjective evaluation and interview of the patients was held prior to every treatment session. Six to eight treatments were required with the alexandrite lasers, 2.4-2.8 months between treatments, and the IPL source required 8-9 treatments, 2-2.5 months apart. RESULTS: No significant difference was seen between the LP- and SP-Alex, or between both of them and the IPL source, although the period to regrowth was longer for the lasers. Erythema and oedema were more noticeable with the LP-Alex, as were crusting and hyper- and hypopigmentation. Discomfort was greatest with the LP-Alex and the IPL source. Hair induction at the borders of the treated area on the face and neck was seen only with the LP-Alex, and correlated statistically significantly with any episode of severe erythema, crusting or hyperpigmentation. CONCLUSIONS: There was no statistically significant difference between the LP-, SP-Alex and IPL photoepilation with regard to efficacy. Transient side effects were highest with the LP-Alex, and least with the IPL system. In the LP-Alex treated face and neck sites, 3.1% had hair induction in the borders of the treated areas. PMID- 14741817 TI - Hair reduction: 48 months of experience with 800nm diode laser. AB - BACKGROUND: The goal of hair removal methods has always been permanent or at least long-term epilation. Laser technology and the axiom of selective photothermolysis allowed a new approach to this subject. OBJECTIVE: The aim of this retrospective study was to determine the long-term efficacy of a solid state, 800nm pulsed near infrared diode laser system for the reduction of pigmented hair, and extending habitual hair plucking intervals. METHODS: In 48 months 242 patients received 703 diode laser treatments on 477 sites. Treatment data and an anonymous patient questionnaire on data concerning undesired hair growth (e.g. predisposition, psychological aspects), and hair plucking habits were retrospectively evaluated to verify the benefit of 800-nm diode laser application for the reduction of excess hair. RESULTS: After an average of 1.97 treatments (range 1-6) sufficient reduction of pigmented hair was achieved for a mean period of 8.1 months. The habitual hair plucking interval was raised from a mean of 3.69 days before treatment to 15.19 days after laser epilation. The procedure was tolerated by 81.4% without local anesthetics and well accepted by 82.55% of the patients. CONCLUSION: 800-nm diode laser assisted hair removal represents a safe and effective treatment option for the reduction of pigmented hair. PMID- 14741818 TI - Ablation of cutaneous lesions using an erbium:YAG laser. AB - BACKGROUND AND OBJECTIVE: This study was performed to evaluate the effectiveness and safety of erbium:YAG laser in removal of cutaneous lesions. STUDY DESIGN: Data were collected after removing 363 benign, pre-malignant and malignant lesions in 27 patients at a dermatology and cosmetic laser surgery center. RESULTS: All lesions were completely removed. Eight of 363 lesions recurred and the histological analysis showed complete removal of one malignant lesion with erbium:YAG laser ablation. There were no long term or permanent complications. CONCLUSION: Erbium:YAG laser is safe and effective in removal of cutaneous lesions. PMID- 14741819 TI - Response of spider leg veins to pulsed diode laser (810 nm): a clinical, histological and remission spectroscopy study. AB - BACKGROUND: Spider leg veins are common. Their treatment with laser or intensed light therapy shows generally variable success rates and often adverse side effects such as hyper- or hypopigmentation. This study was performed to investigate whether pulsed diode laser (810 nm) treatment is effective and safe. METHODS: Thirty-five female patients with spider leg veins were included in this prospective trial. They were treated twice with a pulsed diode laser (810 nm; spot size 12 mm, frequency 2-4 Hz, pulse width 60 msec, fluence 80-100 Jcm(-2)). Laser therapy was performed on day 0 and day 14. Clinical assessments were carried out before and immediately after the first laser therapy, after 2 weeks, 8 weeks, and one year. Skin biopsies were taken before and immediately after the first laser treatment, and after 10 weeks. Contact-free remittance spectroscopy was performed before laser treatment, immediately after the first treatment, after 2 weeks and 8 weeks. RESULTS: After the first treatment 15 patients showed a complete disappearance (CR) of spider leg veins; in the remaining 20 patients a remarkable improvement (RI) was noted (n=35). After six months of follow-up CR was seen in 6 patients, RI in 6, a stable situation in 9, and scar formation in 1 patient (n=21). The effect was almost completely stable during one year of follow up. The examination of histological specimens before and after laser treatment showed no cellular inflammatory reaction. The mean vascular area was significantly reduced after the first (p<0.05) and after the second (p<0.05) laser treatment. Spectral analysis showed a marked decrease of peaks for oxygenized haemoglobin immediately after laser treatment and during the follow up. Safety profile was excellent without purpuric reaction or pigmentary changes. Mild scarring was observed in two patients at the end of follow-up. CONCLUSIONS: Pulsed diode laser therapy (810 nm) is an effective and safe treatment option for spider leg veins. The effects can be seen immediately. Objective monitoring by non-invasive remission spectroscopy and histology of biopsy specimens demonstrates selectivity of the laser action. PMID- 14741820 TI - Hair removal with a combined light/heat-based photo-epilation system: a 6-month follow-up. AB - BACKGROUND: A variety of lasers and light sources have been used for hair removal. A novel flashlamp-based device, with a mechanism of action mixing a combination of light and heat, has recently become available. OBJECTIVE: To assess the efficacy, side effects and long-term results of this new light/heat based system for photo-epilation. METHODS: A total of 21 patients (six male, 15 female), skin types II and III, with dark, coarse hair mainly on the back or chest for males and the bikini/axilla area for females, were reviewed after having been treated during the previous 18 months with this system. Each patient had at least four sessions with a maximum of nine. No depilatory treatment except shaving was allowed during the time of follow-up. The percentage of hair reduction was measured clinically or by comparing "before and after" photographs. RESULTS: After four to nine sessions, depending on the anatomic location, all patients showed hair reduction as well as hair thinning. The percentage of reduction varied depending on the anatomic location and number of sessions, the bikini/axilla area responding better than the back. Transient side effects were limited to a stinging sensation during the session and to redness, which lasted for up to 2 days. No chronic side effects were observed. CONCLUSION: The present study demonstrates that this device is safe and effective for hair removal with a long-term effect. PMID- 14741821 TI - Long-term clinical results of IPL photorejuvenation. AB - BACKGROUND: Non-ablative photorejuvenation is characterized by the reduction of intrinsic and extrinsic changes in photodamaged skin. Only short-term improvement has been documented previously. OBJECTIVE: To evaluate quantitatively the short term and long-term clinical effectiveness of multiple full-face IPL treatments for non-ablative facial photorejuvenation. METHODS: A total of 47 patients with varying degrees of photodamaged skin and rosaceal dermatitis underwent a series of four to five IPL treatments with a Vasculight (Lumenis Corp). Treatments were conducted every 3-4 weeks. Photographs were taken at baseline and after the treatment series was completed at both 6 weeks and 6 months. Adverse effects and clinical improvement were documented. A patient satisfaction questionnaire was completed and reviewed at the 6-month evaluation period. RESULTS: Standardized evaluation of rhytids showed a statistically significant improvement in wrinkles at both evaluation endpoints. Some degradation occurred over time. Facial vascularity, dyschromia, and large pore size progressively improved from the 6 week measurement to the 6-month measurement. CONCLUSION: This clinical study demonstrates that non-ablative facial rejuvenation is associated with long-term clinical improvement of facial rhytids, abnormal vascularity and pigmentary disorders with minimal risks and side effects to the patient. PMID- 14741822 TI - Conversion of aesthetic lasers and intense pulsed light sources into inherently eye-safe units. AB - BACKGROUND: Laser systems used in aesthetic treatments pose a serious risk to the eyes of bystanders and equipment operators. Although much safer than lasers, intense pulsed light (IPL) sources are also not without risk. OBJECTIVE: To present a technology that can convert most lasers and IPL units used in aesthetic clinics into inherently eye-safe devices not requiring the use of protective eyeglasses, thus making them Class I devices. With the exception of Q-switched lasers, this new technology also dramatically reduces the potential hazards of lasers that do not attain Class I level to below that attributed to flashlamps. A device that eliminates the annoying glare generated by the reflection of pulsed light from the skin is also presented. MATERIALS AND METHODS: The new eye-safe technology is based on the attachment of a highly efficient wide-angle forward scattering optical diffuser to the distal end of the laser delivery handpiece. This device is used in contact with the treated area without modification of the conventional treatment parameters and preserves clinical efficacy. The handpiece is designed to act as an eye-safe, extended diffusing light source complying with Class I eye safety standards in most laser types. The technology also makes IPL devices safer by reducing the radiance of the flashlamp, which is hazardous when viewed directly by operators or bystanders. Skin glare is also eliminated by a liquid crystal, glare-free window activated in synchronization with the treatment pulses. RESULTS: Clinical hair removal results with an 80 degree diffuser did not reveal any difference when compared with results obtained with the original laser source. The incorporation of a diffuser in an IPL unit has not modified its clinical efficacy. CONCLUSION: With the exclusion of Q-switched lasers, it is possible to convert a large number of lasers and IPL units utilized in aesthetic treatments into inherently eye-safe units, thereby considerably enhancing the comfort and safety of aesthetic clinics without sacrificing clinical efficacy. PMID- 14741823 TI - Delayed healing after CO2 laser resurfacing. AB - Skin resurfacing with carbon dioxide (CO(2)) laser is a currently popular means of improving rhytides and scars of the face. Side effects are few, but delayed healing has been reported due to infections. A case of unexplained slow healing following CO(2) laser resurfacing in a previously healthy sixty-seven year old woman is reported. Although healing was extremely delayed, it is important to emphasize that a satisfactory cosmetic response was obtained after 16 months. PMID- 14741824 TI - Long-term hypopigmentation induced by diode laser photo-epilation. PMID- 14741825 TI - Proceedings of the 2003 International Master Course on Aging Skin. Paris, France. PMID- 14741826 TI - Epilation with Nd:YAG laser: a brief analysis of the technical application methods, results and pre- and post-treatment procedures. AB - With the use of the so-called "selective" lasers, it is possible to operate specifically on the melanin as the absorbing target of undesirable hair. The optimal pulse duration for laser epilation should be found between the TRT of the epidermis (1-2 msec) and the TRT of the follicle (10-50 msec). For the thermal destruction of the follicle containing melanin, the ideal laser energy should (1) be absorbed in a selective manner with respect to other chromophores; (2) have a wavelength capable of penetrating in depth and reaching the target to be treated; and (3) be sufficient to destroy tissue targets (in the hair the papilla, the follicular epithelium, the so-called "bulge" and also the vascular support). With this method the melanin and the follicular structures are "heated" in a preferential manner while still safeguarding the adjacent skin structures. Numerous laser systems are employed in the treatment of undesirable hair with varying results according to the source, procedures, phototype, and pigmentation of the hair to be treated. The most frequent side effects may be discomfort or a slight burning sensation and un-aesthetic pigment changes or scarring. We report here our experience based on the treatment of 94 subjects over a 12 month period using the Nd:YAG laser (SMARTEPIL 2, Deka M.E.L.A., Italy). Our results indicate that a wavelength of 1064 nm is efficacious in obtaining a greater penetration and reaching the deeper follicles, with a greater respect for the skin due to the lower absorption by the melanin, and consequently a lower incidence of side effects. PMID- 14741827 TI - Laser leg vein treatment: a brief overview. AB - Laser treatment of leg veins has been associated with a number of disadvantages, but the introduction of new devices has increased the role of lasers in the treatment of leg veins. This paper reviews the role of laser devices applied from the surface in the treatment of reticular and spider veins. Success is determined by the proper selection of wavelength, fluence, pulse duration, spot size, and number and frequency of treatments. PMID- 14741828 TI - The use of lasers and intense pulsed light sources for the treatment of acquired pigmentary lesions in Asians. AB - Asians are prone to the development of lentigines as a result of photoageing. Lasers and intense pulsed light sources have been used in the treatment of lentigines but there is a risk of post-inflammatory hyperpigmentation in Asian skin. This report examines the role of lasers and intense pulsed light sources in treating lentigines and other acquired pigmentary lesions in Asians. PMID- 14741829 TI - ED2000: 585 nm collagen remodelling pulsed dye laser. AB - The wavelength of 585nm corresponds to an absorption peak of haemoglobin. The heating effect in these skin layers triggers the release of various growth factors that stimulate collagen remodelling and tightening. We report our experience with a 585nm collagen remodelling, double flashlamp excited pumped dye laser was used (ED2000, Deka MELA, Calenzano, Italy), spot size 5 mm, energy density (fluence J/cm(2)) from 2 to 4 J/cm(2), emission modality (repetition rate) at 0.5 Hz, with a short pulse duration of 250 microsec. The efficiency of 585 nm collagen remodelling pulsed dye laser is controversial in only one session. It is probably reasonable to inform patients that 3-4 treatment sessions are necessary, and that 10% of the patients have no response to nonablative photorejuvenation. Because of its low fluence and its shorter pulse duration, the 585 nm collagen remodelling pulsed dye laser has limited efficacy for the treatment of port wine stains. However, it may offer patients with erythematous, raised or hypertrophic acne scars or striae distensae a permanent cosmetic solution. This laser is safe and effective in the treatment of surgical scars starting as soon as possible, on the day of suture removal if possible. We found that 96.3% of molluscum contagiosum healed after the first treatment, the other 3.7% after the second. PMID- 14741830 TI - The 755 nm alexandrite laser. PMID- 14741831 TI - New technology in aesthetic medicine: ELOS electro optical synergy. PMID- 14741832 TI - Photorejuvenation using intense pulsed light: my technique. PMID- 14741833 TI - Worldwide experience with botulinum toxin type A. PMID- 14741834 TI - Botulinum toxin in association with other rejuvenation methods. AB - Botulinum toxin is an import agent in facial rejuvenation, and the use of botulinum toxin in combination with other methods helps preserve and maintain a long-term result. This paper briefly reviews the use of botulinum toxin in associate with dermal fillers, laser resurfacing and other surgical rejuvenation modalities. PMID- 14741835 TI - Challenges in the mid and the lower face. AB - The cosmetic use of botulinum toxin in the mid and lower third of the face has some challenges that must be overcome with experience. This paper briefly reviews the application of botulinum toxin in these regions. The treatment of the mid and lower face with botulinum toxin should be carried out with low doses. When starting with this technique, it is advisable to do a two-step treatment especially for those not acquainted with these regions. PMID- 14741836 TI - Use of botulinum toxin in facial paralysis. AB - Facial paralysis triggers aesthetic and functional changes that hold significant physical and psychological repercussions to those suffering from this condition. Botulinum toxin may be used to reduce the facial asymmetry encountered in patients suffering from facial paralysis. The onset of improvement starts usually 15 days after the injection. Patients' satisfaction increases with time, especially after 30 days. Adverse events concerning the perioral area are found but resume as the patients get used to the new dynamics. The use of botulinum toxin is effective in decreasing the hyperkinesis contralateral to facial paralysis. The global aesthetic aspect of the face improves. PMID- 14741837 TI - Botulinum toxin dilution: our technique. AB - The optimal dilution of botulinum toxin remains a controversial matter. With the use of botulinum toxin in new and more delicate indications, dilution is the main procedure to reduce side effects. The objective of this paper is to review the literature and report our dilution method and our experience. We use either 500 U Dysport and 100 U Botox sterile vials. For facial indications we dilute them with 1 ml of sterile solution and in Dysport solution we add adrenalin with final solution 1:100.000. For platisma and dysidrosis we use more diluted solutions without adrenalin. It is very important to use a 30 U syringe to properly calibrate the low doses used. Although there is a great variation in toxin dilution used by practitioners, in our opinion this particular aspect of the technique remains very important: using low amounts of toxin in very low amounts of saline solution reduces side effects. The use of adrenalin, such as for local anesthetic solutions, needs more study but may be useful to reduce doses, to minimalize spread in nearby muscles and, in our experience, does not produce side effects. PMID- 14741838 TI - How to avoid brow ptosis after forehead treatment with botulinum toxin. AB - Botulinum toxin is primarily indicated on the upper third of face. The treatment of the forehead wrinkles has a limited number of side effects, but can lead to brow ptosis. Brow ptosis appears in many studies with a frequency of around 1-5%. This is caused, often, by using incorrect injection sites, too high dosages, and by an inappropriate selection of patients. In our experience, this side effect has occurred in less than 1% of cases. This paper will emphasize the technical methods we use in order to reduce to a minimum this unpleasant side effect. Understanding the side effects of botulinum toxin on forehead requires a thorough understanding of muscular anatomy. We emphasize the importance of the correct dilution, the use of adrenalin and a 30 U syringe, particularly for patients who have skin abundance (dermatocalasys) of forehead. Patients with dermatocalasys should usually be excluded, but if treated, it should be done in two different sessions, without overdosing and respecting the symmetry. Brow ptosis, the only real side effect of forehead treatment with botulinum toxin A, can be reduced to a minimum by using simple rules. PMID- 14741839 TI - Full face endolift: 10 years evolution. AB - The author describes his personal procedure for endolift centred on the midface. Techniques are described for superior blepharoplasty, dissection of the midface and subperiosteal forehead lifting. The suspensions are multiplied and matched to each case selection. The neck is treated according to Giampapa's technique. The results are stable over a five-year follow-up. This technique has been significantly simplified; recovery is rapid and the risk of injuring of the branches of the facial nerve is dramatically reduced. PMID- 14741840 TI - Bio-Alcamid in drug-induced lipodystrophy. AB - The development of new drugs to counter human immunodeficiency virus (HIV) infection has led to an increase in lipodystrophic syndrome among HIV-infected individuals receiving combination therapy. Bio-Alcamid(TM) is a recently developed polymeric substance that can be implanted to compensate for adipose effects. We have implanted this substance in 73 patients with up to three years' follow-up. The aesthetic results were deemed excellent by both physicians and patients. No implant dislocation, implant migration, granuloma, allergic reaction or intolerance were recorded. PMID- 14741841 TI - European breast implant registry--plastic surgery center: European strategic coaching. PMID- 14741842 TI - Fibroblasts: what's new in cellular biology? AB - This paper briefly examines the fibroblast network with particular emphasis on the exceptionally complex pattern of specific interactions and their effects on dermal integrity and homeostasis regulation systems. It will be some time before we have a full understanding of the cellular biology mechanisms involved in the operation of lasers, flashlamps, peels, mechanical dermabrasions, fillers or topicals on the skin. PMID- 14741843 TI - Aminopeptidases and angiogenesis. AB - A number of proteases, including matrix metalloproteinases and plasminogen activators, have been shown to be involved in angiogenesis. In addition, recent reports suggest that aminopeptidases also play roles in angiogenesis. These peptidases regulate the N-terminal modification of proteins and peptides required in processes such as maturation, activation, or degradation, and thereby they are related to a variety of physiological and pathological processes. At least three aminopeptidases are reported to be involved in angiogenesis, namely, type 2 methionine aminopeptidase, aminopeptidase N, and adipocyte-derived leucine aminopeptidase/puromycin-insensitive leucyl-specific aminopeptidase. This review will focus on the possible role of these aminopeptidases in angiogenesis. PMID- 14741844 TI - Endothelial NADPH oxidase: mechanism of activation by low-density lipoprotein. AB - Exposure to atherogenic levels of low-density lipoprotein (LDL) causes elevated reactive oxygen species (ROS) production by human endothelial cells (ECs). NADPH oxidase is thought to be the main source of ROS generated by LDL-activated ECs. The mechanism by which this lipoprotein activates endothelial NADPH oxidase is incompletely understood. To gain further insight into the signaling pathway, the authors have examined the effects of inhibitors to various signal transducing enzymes, including the G(i)-protein coupled receptor (pertussis toxin), Src tyrosine kinase (PP1), phospholipase C-gamma (U73122), phosphatidylinositol 3 kinase (LY294002), p42/p44 mitogen-activated protein kinase (MAPK) kinase (PD98059), p38 MAPK (SB203580), protein kinase C (Ro 318220, GF 109203X, Go 6976), and cytosolic phospholipase A(2) (AACOCF3), on the ROS-producing capacity ECs activated by LDL. Exposure of cultured ECs to LDL (0.45 mg protein/mL) stimulated ROS formation, as measured using a 6-carboxy-2',7' dichlorodihydrofluorescein diacetate assay. This effect was partially inhibited by Ro 318220, GF 109203X, U73122, and SB203580, and blocked or nearly completely inhibited by PP1, pertussis toxin, LY294002, PD98059, and AACOCF3. Only a partial, minor inhibition occurred with the protein kinase C inhibitor, Go 6976. These results are most consistent with LDL activating endothelial NADPH oxidase, predominantly through a signaling pathway that leads to cytosolic phospholipase A(2) activation. PMID- 14741845 TI - Interferon (IFN)-beta 1a and IFN-beta 1b block IFN-gamma-induced disintegration of endothelial junction integrity and barrier. AB - Recent clinical trials indicate the efficacy of interferon (IFN)-beta 1b in reducing relapse rate in relapsing-remitting multiple sclerosis (MS), whereas a surge of IFN-gamma precedes and provokes acute relapses. Disruption of the cerebral endothelial barrier and transendothelial migration of inflammatory cell migration into the brain play a significant role in pathogenesis of MS and may be driven by this surge in IFN-gamma. However, the molecular mechanisms underlying the beneficial effects of IFN-beta 1b against the deleterious effects of IFN gamma on the barrier formed by the junctional proteins remain to be characterized. The authors investigated the effects of IFN-beta 1b, IFN-beta 1a, and IFN-gamma on the integrity of two endothelial junctional proteins, occludin and vascular endothelial-cadherin (VE-cadherin). Human umbilical vein endothelial cell (HUVEC) layers were treated with IFN-beta 1b, IFN-beta 1a, IFN-gamma, IFN beta 1b plus IFN-gamma, or IFN-beta 1a plus IFN-gamma. IFN-beta 1b, IFN-beta 1a, and IFN-gamma effects on occludin and VE-cadherin integrity and electrical resistance were assessed by Western blotting and immunofluorescence. IFN-gamma significantly reduced occludin expression and produced gaps in endothelial monolayers. VE-cadherin expression was decreased to a lesser extent in endothelial cells exposed to IFN-gamma. IFN-beta 1b significantly attenuated the IFN-gamma-induced decrease in occludin and VE-cadherin expression. The protective effects of IFN-beta 1a on IFN-gamma-treated endothelial cells were similar to those of IFN-beta 1b. IFN-gamma also significantly reduced endothelial monolayer electrical resistance; this effect was blocked by either IFN-beta 1a or IFN-beta 1b. IFN-beta 1a and IFN-beta 1b effectively prevent the IFN-gamma-induced disintegration of the endothelial tight junctions and sustain barrier against the effects of IFN-gamma. The protective effects of IFN-beta on occludin and VE cadherin stability appear to represent molecular mechanisms for the therapeutic effects of the IFN-beta on blood brain barrier in MS. PMID- 14741846 TI - Effect of cGMP on lung microvascular endothelial barrier dysfunction following hydrogen peroxide. AB - The authors determined the effect of cyclic guanosine 3',5'-monophosphate (cGMP) on hydrogen peroxide (H(2)O(2))-induced barrier dysfunction in bovine lung microvascular endothelial cell (BLMVEC) monolayers and compared the results to bovine pulmonary artery endothelial cells (BPAECs). In BLMVECs, H(2)O(2) (250 microM) caused a 31.9% +/- 4.8% decrease in transendothelial electrical resistance (TER) associated with increased actin stress fiber formation, intercellular gaps, and intracellular calcium concentration ([Ca(2+)](i)). The cGMP analogue 8-(p-chlorophenylthio)-cGMP (8p-CPT-cGMP; 30 or 50 microM) prevented the H(2)O(2)-induced decrease in TER (p <.001) as well as the cytoskeletal rearrangement and intercellular gap formation. 8-pCPT-cGMP (50 microM) attenuated the peak (418.8 +/- 42.1 versus 665.2 +/- 38.0 nmol/L; p <.001) and eliminated the sustained increase in [Ca(2+)](i) (193.5 +/- 21.3 versus 418.8 +/- 42.1 nmol/L; p <.001) caused by H(2)O(2). 8-pCPT-cGMP also increased TER (14.2% +/- 2.2%; p <.05) and decreased [Ca(2+)](i) (201.2 +/- 12.5 vs. 214.4 +/- 12.1 nmol/L; p <.03) before H(2)O(2). In BPAECs, 8p-CPT-cGMP significantly attenuated H(2)O(2)-induced increases in permeability and [Ca(2+)](i) but less effectively than in BLMVECs. These results suggest that in BLMVECs, cGMP countered the adverse effects of H(2)O(2) on barrier function by preventing actin cytoskeletal rearrangement and attenuating the increase in [Ca(2+)](i). PMID- 14741847 TI - Establishment and characterization of an angiosarcoma-derived cell line, AS-M. AB - A novel human endothelial cell line, AS-M, has been established from a cutaneous angiosarcoma on the scalp. The cells expressing platelet endothelial cell adhesion molecule-1 (CD31) were isolated using magnetic beads and subsequently cultured for a year. To date, the cells have undergone more than 100 population doublings (PDs). The AS-M cells manifested endothelial characteristics, such as active uptake of acetylated low-density lipoprotein labeled with 1,1'-dioctadecyl 3,3,3',3'-tetramethylindocarbocyanine perchlorate (Dil-Ac-LDL), capacity to bind the Ulex europeaus agglutin-I (UEA-I), and expression of von Willebrand factor (vWF) and CD31. The single cell-derived clone, AS-M.5, showed a constitutive expression of CD31, vWF, angiotensin-converting enzyme (ACE), endoglin (CD105), and the endothelial cell receptor tyrosine kinases KDR and Tie-1. Similarly to freshly isolated endothelial cells, the AS-M.5 responded to induction by bacterial lipopolysaccharide (LPS) by increased transcription of cell adhesion molecules and cytokines. The AS-M.5 cultures required endothelial growth supplements for optimal growth and long-term propagation in vitro. However, in contrast to normal endothelial cells, p53 gene products were detected in nuclei of AS-M.5 cells. Cytogenetic analyses consistently revealed a hypodiploid karyotype with complete loss of one homologue of several chromosomes and a homogeneous pattern of distinct karyotypic changes. Although the AS-M.5 presented characteristics suggestive of tumor cells, they did not develop into tumors when inoculated subcutaneously into nude mice. The cell line AS-M.5 could be a useful model system to study endothelial pathobiology in vitro. PMID- 14741848 TI - Purified and proliferating endothelial cells derived and expanded in vitro from embryonic stem cells. AB - Embryonic stem (ES) cells serve as an excellent in vitro system for studying differentiation events and for developing methods of generating various specialized cells for future regenerative therapeutic applications. Two obstacles associated with using embryonic stem cells include (a) isolating homogeneous populations of differentiated cells and (b) obtaining terminally differentiated cell populations that are capable of proliferating further. Here, the authors describe methods in which they have overcome these two obstacles by generating highly purified populations (>96%) of actively proliferating endothelial cells from mouse ES cells. Briefly, 60,000 ES cells progress through three different stages of cell induction/expansion and two cell isolation procedures, generating over 300 million endothelial cells. These ES-derived endothelial cells display characteristics similar to vascular endothelial cells in that they express several common endothelial markers, they form two-dimensional (2D) tubelike structures as well as complex microvessels in three-dimensional (3D) collagen type I gels, and they retain the ability to reorganize their cytoskeleton in response to mechanical forces. Our findings indicate that it is possible to obtain proliferating populations of homogeneous endothelial cells from mouse ES cells without genetically manipulating the ES cells or coculturing with feeder cells. PMID- 14741852 TI - Developing the teaching instinct 5: mentoring. AB - Mentoring is an activity that has many different interpretations. It is usually seen as a relationship that encourages the holistic development of a person. This means that the mentor role encompasses being an exemplary role model, a confidante who will support the mentee through any personal stresses and strains, a teacher, a developer of talent and an opener of doors. PMID- 14741851 TI - Ethics assessment in medical students. PMID- 14741853 TI - E-learning in sexuality education. AB - The use of computers in learning and communication is not new to health professional education. However, the rapid developments in technology and the increasing competence of students in computer- and web-based learning make the need for health professional education to take the next step to e-learning a necessity rather than a choice. This paper describes an e-learning unit in sexuality developed in response to a felt need for a flexible online unit in the topic. The use of online tools for e-learning in sexuality are discussed and course evaluation presented. The need for a strong pedagogic model and the careful development of learning activities to utilize the facilities available for assessment, feedback and especially synchronous and asynchronous communication are discussed as they apply to the sexuality unit. PMID- 14741854 TI - Use of portfolios in early undergraduate medical training. AB - The ability to reflect on one's own action is seen as an important skill for a doctor. A thorough introduction of the portfolio planned in the early stages of their studies seems to be the way to train medical students in reflection. This article describes the use of portfolios in early undergraduate medical training. The literature on portfolios suggests three aspects that are crucial for the effectiveness of portfolios: structure, coaching and assessment. The portfolio system was designed by transposing the experience with portfolio systems outside and inside medical training to a situation of first-year medical students. During the academic year 2001-02 242 first-year medical students compiled a portfolio. Student experience was collected by semi-structured interviews. The majority of students were of the opinion that analysing one's competences in a portfolio was instructive and meaningful. With regard to learning how to reflect and recognize learning needs, however, mentor coaching proved to be necessary. The results thus far show that the portfolio is a worthwhile addition to existing assessment and learning tools. PMID- 14741855 TI - How strong is the case for the adoption of problem-based learning in physiotherapy education in the United Kingdom? AB - Despite its use in medical and health education in several countries, problem based learning (PBL) has to date not been adopted in undergraduate physiotherapy education in the UK. Most of these physiotherapy curricula incorporate many of the educationally desirable features found in PBL. The question is whether the evidence for PBL is strong enough to justify the costs involved in adopting it on such programmes. The strength of the evidence for PBL was reviewed in relation to required graduate attributes and several aspects of learning theory. Overall the findings were equivocal. The evidence suggests that PBL is preferential to traditional curricular design, but does not seem to meet all the requirements of medical and health education. Adoption of PBL would be positive, but its processes and outcomes may be no more successful than those of other curricular designs that incorporate the same sound educational design features. Comparative research in this area is needed. PMID- 14741856 TI - Predicting academic success by group behaviour in PBL. AB - The aim of this study was to explore predictors of medical students' academic success by variables covering group behaviour, individual work and personality measures. The working hypothesis was that peer evaluation of behaviours in group work was of major importance. First-year medical students (n=199) at the Norwegian University of Science and Technology were included. All variables were derived by questionnaires and tested in relation to academic success in the written exam at the end of the first year. In multiple regression analyses, the group work behaviour 'improving learning' emerged as a significant predictor together with 'limiting good group process'. In the bivariate analyses, individual work variables such as constructive learning styles were also found to be related to academic success. External 'locus of control' was negatively associated with constructive learning styles and positively with dysfunctional ones. PMID- 14741857 TI - Evaluation of a formative interprofessional team objective structured clinical examination (ITOSCE): a method of shared learning in maternity education. AB - Shared learning at undergraduate level provides a potential means of promoting a more multi-professional approach to maternity care. Interprofessional education uses shared interactive sessions to promote different professional groups' understanding of each other and working together. This paper describes the use of a formative objective structured clinical examination as a method of interprofessional education. Mixed groups of student doctors and student midwives rotate through a series of clinical stations based on common labour ward scenarios. After completing each station they are given feedback by a facilitator on their problem-solving skills, knowledge and attitude to team working. The interactive nature of the sessions encourages deep learning, is student centred and promotes a positive attitude to multidisciplinary working. Both student groups felt they benefited from shared learning in this way and that the formative OSCE was an effective method of developing their clinical skills. PMID- 14741858 TI - Was a breach of examination security unfair in an objective structured clinical examination? A critical incident. AB - One-third of a class of students was inadvertently given the names of stations immediately prior to an OSCE and two-thirds of the class were not. This provided an opportunity to explore student perceptions of fairness and to explore any effect of this cueing. The subjects were medical students undertaking an end of fifth year multidisciplinary OSCE. OSCE score data from the 20 students who had received the information were compared with those of the 40 students who did not. We also compared their performance on other assessments to determine whether the two groups were comparable. The overall OSCE mark was not significantly different between the two groups. There were significant differences between groups on four stations but this was not in a consistent direction that advantaged one group. There were no significant differences between the two groups in their performance on the other examinations. This inadvertent security breach had no systematic effect on student OSCE station scores. This incident provided a valuable opportunity to admit error, approach it rationally and restore any resulting breach of trust. PMID- 14741859 TI - Health science students' time organization and management skills: a cross disciplinary investigation. AB - One hundred and fifty-four final year health science undergraduates from the disciplines of Medical Radiation Sciences, Speech Pathology and Physiotherapy in the Faculty of Health Sciences, the University of Sydney completed the Australian Time Organisation and Management Scale (ATOMS). Findings identified the strengths and weaknesses of students on the four dimensions included in this analysis. Whilst the results indicated strengths in the area of students' sense of purpose, level of focus and goal-setting ability, areas of weakness were noted, particularly with regard to the use of basic time-management devices (e.g. use of a diary and making lists). How these skills and competences are related to age, gender and discipline is explored. In addition, the degree to which these time management behaviours can be enhanced (eg with time management intervention programs), the generalizability of the findings to other settings (non-academic), and the development of these skills over time are major issues discussed in the paper. PMID- 14741860 TI - Embracing the new paradigm of assessment in residency training: an assessment programme for first-year residency training in anaesthesiology. AB - Assessment of clinical competence is facing a paradigm shift in more than one sense. The shift relates to test content, which increasingly covers a broader spectrum of competences than mere medical expertise, and to test methods, with an increasing focus on testing performance in realistic settings. Also there is a shift in the concept of assessment in that instruction and assessment are no longer seen as being separate in time and purpose, but as integral parts of the learning process. The nature of the new paradigm for assessment is well described but the challenge to programme directors is to specify the evaluation situations and develop appropriate methods. This paper describes the intrinsic rational validation process in outlining an assessment programme for first-year anaesthesiology residency training according to the new paradigm. The applicability to other residency programmes and higher-level training in anaesthesiology is discussed. PMID- 14741861 TI - Introducing performance-based assessment of family physicians. AB - Slovenia has implemented a new final summative assessment for vocational training in family medicine. As all family medicine trainees work independently in practice, the assessment is focused on actual performance. The preparation included a literature search, consultation with foreign experts, consultation with tutors and trainees in general practice and testing the assessment with experienced practitioners. The assessment includes three elements: (1) a trainee's report with data on population served, audit of practice and family profiles; (2) a visit to the trainee's practice which includes sitting in with the trainee, and assessment of patients' records, equipment and premises; (3) the final assessment, which includes an MCQ written test, OSCE stations, and an oral exam with discussion about the candidate's report, presentation of five cases from his/her practice and one question by each examiner to assess problem-solving skills. Initial experience has been favourable but further work is needed for evaluation of the assessment. PMID- 14741862 TI - New doctors' perceptions of their educational development during their first year of postgraduate training. AB - The General Medical Council (GMC) in the UK describes a wide range of competences in which the new medical graduate should develop during the first postgraduate year. New graduates are known as pre-registration house officers (PRHOs). In this study, semi-structured open interviews with PRHOs approaching the end of their first postgraduate year were used to explore the trainees' perceptions of their development. After the interview their views on the importance given in their experience to the GMC's competences were explored though a questionnaire. Most noticeable from the interview data was the PRHOs'emphasis in the development of generic skills required of working in the 'real world'--skills such as communication, team working and the ability to cope with responsibility. In some outcomes, their noticeable progression, e.g. communication, or lack of progression, e.g. health promotion, reflected their views on the importance given. With other outcomes there was a paradox, e.g. practical skills were rated highly but were not noticeably developed; the role of the doctor was noticeably developed but was not highly rated. Overall the PRHOs developed most strongly in the 'professionalism' domain with not all the skills in the 'tasks' domain being similarly developed. This study helps to define the benefits and opportunities of the PRHO year, as currently perceived by the trainees, and should be of interest to those involved in the planning and delivery of postgraduate training. PMID- 14741863 TI - A systematic review of postgraduate teaching in evidence-based medicine and critical appraisal. AB - The knowledge and skills needed for critical literature appraisal and evidence based practice have not been covered in undergraduate education until recently. These educational needs are, therefore, often met through postgraduate education via courses, workshops and journal clubs. Previous reviews have generally considered postgraduates and undergraduates together. However, there is evidence that the effectiveness of educational interventions varies between postgraduates and undergraduates. In this study the authors therefore examine the effectiveness of evidence-based medicine and critical appraisal teaching at postgraduate level. A comprehensive search was conducted in MEDLINE, EMBASE, ERIC, CCTR, CDSR, DARE, HTA, Best Evidence and SCI. Seventeen studies were identified: two randomized trials, six controlled trials without randomization and nine before-and-after studies. The studies showed a significant improvement in knowledge, but not in attitude, skills or behaviour. However, to draw robust conclusions, well-designed trials are needed that focus on curriculum content and delivery as well as how outcomes are assessed. PMID- 14741864 TI - Reflective critique and collaborative practice in evaluation: promoting change in medical education. AB - The School of Medicine at the University of Tasmania has recently begun to implement a process of curriculum evaluation,which aims to reflect contemporary best practice in evaluation in tertiary pedagogy and medical education. Best practice must accommodate a broadening interest in cooperative and collaborative evaluation strategies among stakeholders, advances in applied qualitative educational research and recognition that critical reflection on practice is the cornerstone of professional education. This paper reports a recent evaluation strategy in a specific year-long unit in the second year of a six-year undergraduate medical degree. The paper begins by presenting the context; it then discusses and rationalises the evaluation strategy and presents findings. The paper concludes by arguing that curriculum evaluation as best practice must be reflective, informed by the scholarship of medical education, and internalized as a dynamic process that can promote sustainable change and improvement in medical curricula. Such an approach will contribute to an undergraduate medical curriculum that prepares students for the demands and complexities of medical practice. PMID- 14741865 TI - A transitional course from high school to medical school in a new medical curriculum in Asia: how do the students see it? AB - The objective of this study was to examine the students' views on a transitional course for Asian freshers in a revised medical curriculum. Data were collected through the year-end questionnaire distributed to all first-year students over three academic years from 1997 to 2000. The students considered the transitional course had encouraged them to become more active and self-directed although there were different views about its overall effectiveness. Students appeared uncertain as to the depth in which they were expected to master the subjects, thus leading them to call for more clearly stated learning objectives to help relieve the anxiety they had towards the examinations. Lectures and self-assessment exercises were seen to be providing the general guides for that. PMID- 14741866 TI - Teachers' responsibility: a Socratic dialogue about teaching medical ethics. AB - Old, wise Socrates (SO) is strolling along the Agora to buy some fruit for his wife Xanthippe. He meets Paidagogos (PA),a teacher of medical ethics in the ancient Polis of Athens. Both engage in a discussion on teachers' responsibility and its implications for the design and structure of learning sessions at medical school. PMID- 14741874 TI - The health professional brain drain. PMID- 14741875 TI - Changes in health-related attitude and self-reported behaviour of undergraduate students at the American university of Beirut following a health awareness course. AB - The importance of the university as a setting for health promotion is increasingly being acknowledged. Part of this health promoting function includes curricular offerings to increase health awareness. However, there is a dearth of systematic evaluations of such courses. OBJECTIVE: To evaluate the impact of a university level "Health Awareness" course on attitudes and behaviours of undergraduates enrolled in the course. METHODS: A self-administered survey was used to assess attitude and self-reported behaviour of students at the beginning and end of the course. Paired analysis of means compared responses at pre and post assessments for groups of items within a particular health topic. Cross tabulations of stage of change at pre and post assessment indicated movement related to tobacco use, exercise, and nutrition. RESULTS: Results indicated an improvement of at least 20% from pretest score in four out of eleven health topic areas, and of 10-20% in an additional five topical areas. In addition, movement in a health promotive direction along the stages of change was evident for smoking, eating fruits and vegetables, and exercise. DISCUSSION: The results presented herein are encouraging and indicate support for the impact of a health awareness class on knowledge, attitude, and behaviour of undergraduate students. The two topical areas that did not show improvements between pre and post assessment were those (i). for which students already scored high at pre assessment, or (ii). which the course did not tackle specifically. Conclusions to be drawn are limited by several factors inherent in the design of this evaluation. Future evaluation should include a larger number of students and a comparison group. PMID- 14741876 TI - "Where have all the students gone?" Retaining medical school graduates through educational innovations. AB - CONTEXT: New Mexico is a sparsely populated rural state in the USA, with 20% of New Mexicans living in poverty. There is a need for physicians in the state, especially in primary care. New Mexico's only medical school, the University of New Mexico School of Medicine is state supported. New Mexico and its medical school have a vested interest in its graduates returning to the state to practice. OBJECTIVES: Evaluate the effects of early community preceptorships on graduating physicians' specialty choice and subsequent return to practice in the state where they attended medical school. STRATEGIES: A Primary Care Curriculum (PCC) was introduced into the medical school in 1979. Students expressing interest in this programme were considered. Twenty students per year were assigned to the PCC. The remainder went through the traditional curriculum. The PCC used a problem-based learning model. All PCC students participated in a 16 week, one-on-one community preceptorship (Phase 1B) in a small or rural community during their first year. Graduates from 1983-1996 were analysed for specialty and practice location. MAIN OUTCOMES: Of the 294 students doing Phase 1B, 99 (40%) returned to New Mexico to practice compared to 221 (32%) of traditional students. Fifty-eight (23%) of the Phase 1B students practice primary care compared to 112 (16%) of the traditional students. CONCLUSION: Self-selected students choosing and participating in early community-based clinical experiences coupled with problem-based learning are more likely to return to the state and to practice primary care. PMID- 14741878 TI - Teaching clinical skills in developing countries: are clinical skills centres the answer? AB - CONTEXT: There is growing international interest in teaching clinical skills in a variety of contexts, one of which is Clinical Skills Centres. The drivers for change making Skills Centres an important adjunct to ward and ambulatory teaching come both from within and outside medical education. Educationally, self-directed learning is becoming the accepted norm, encouraging students to seek and maximize learning opportunities. There are global changes in health care practice, increased consumerism and increasing student numbers. In some countries, professional recommendations influence what is taught. Increasingly, core skills curricula and outcome objectives are being defined. This explicit definition encourages assessment of the core skills. In turn, all students require equal opportunities to learn how to practise the skills safely and competently. The moves towards interprofessional education make joint learning in a"neutral" setting, like a Clinical Skills Centre, appear particularly attractive. OBJECTIVE: To discuss the potential role of Clinical Skills Centres in skills training in developing countries and to consider alternative options. DISCUSSION: Many developing countries seek to establish Clinical Skills Centres to ensure effective and reliable skills teaching. However, the model may not be appropriate,because fully equipped Clinical Skills Centres are expensive to set up, staff; and run. They are not the only way to achieve high quality clinical teaching. Suggested options are based on the philosophy and teaching methods successfully developed in Clinical Skills Centres that may fulfil the local needs to achieve low cost and high quality clinical teaching which is reflective of the local health needs and cultural expectations. PMID- 14741877 TI - Career intentions of UNITRA medical students and their perceptions about the future. AB - CONTEXT: The University of Transkei (UNITRA) medical programme follows a problem based learning and community-based education curriculum. OBJECTIVE: To gauge the views of UNITRA medical students about their future, and their career intentions. DESIGN: A semi-structured questionnaire on intentions to specialize or not, preferred place of work, overall view of the future, and careers guidance. FINDINGS: The questionnaire was completed by 364 students (87.7% response rate). The sector preferences are 82.3% public, 6.8% university and 10.9% private. Place of work preferences are urban (37%), rural (27%), city (12%) and abroad (8%), while 16% have no preference. Rural preference varies from 48.5% among 1st year students to 5.9% among 6th year students, while urban preference are 26.2% for Year I and 64.7% for Year VI students. 89.8% of respondents intended to specialize, mainly in clinical fields, with the most influential factors in the choice of specialty being interest and challenge. Most students (78%) view the future positively, 13% say it is uncertain, 8% have no idea about the future; and 1% think the future is negative. Frequent reasons cited for a bright future are job-related, personal attitude, sense of achievement, and the type of training. Most students have received little or no career guidance and would like such topics in the curriculum. CONCLUSION: UNITRA medical students are optimistic about the future. The majority intend to stay in South Africa and work in the public sector, and most of them wish to pursue clinical specialties. PMID- 14741879 TI - Medical students' attitudes towards concordance in medicine taking: exploring the impact of an educational intervention. AB - BACKGROUND AND OBJECTIVES: Concordance has been suggested as a new way of describing the agreement about medicine taking during the consultation process. The aim is a decision on management agreed on by both doctor and patient. As such it has strong links with shared decision-making and patient partnership. In order to encourage doctors to adopt a concordant model, we need to foster a positive attitude towards the concept. We decided to investigate the attitudes of first and second year medical students towards concordance as a base for further educational interventions. SETTING: The School of Medicine, Leeds University. METHODS: We administered the Leeds Attitude toward Concordance scale (LATCon) to first and second year medical students at the beginning of the academic year, and to the same second year students after they had completed a written exercise relating to concordance. RESULTS: The response rate was over 80% for each group. There was no difference in the attitudes towards concordance of the first years and the second years prior to the intervention. After they had completed the exercise, the second years' attitudes towards concordance improved by a small but significant amount. CONCLUSIONS: A paper-based exercise with questions focusing on concordance and based around cases appears to improve medical students' attitudes towards the concept. This exercise needs to be followed up with skills training and observation of role models in order that the attitudes of the students translate into practice once they are qualified. PMID- 14741880 TI - A conceptual model for empowerment of the female community health volunteers in Nepal. AB - CONTEXT: The existing top-down approach of the training is not appropriate to empower the community health volunteers to produce a sustainable change in their performance. OBJECTIVE: To propose a conceptual model that relies on Freire's theory of empowerment education and the participatory action research methodology to empower the female community health volunteers (FCHVs) to increase their consciousness, competence and confidence in performing their job responsibilities. METHOD: The model explains the empowerment phenomenon as a process and outcome. As a process it occurs in the form of repeated reinforcement cycles with alternating activities of action and reflection and includes developing awareness, skills and confidence among FCHVs through small group activities. As an outcome, it results in the change in FCHVs' performance in increasing contraceptive acceptance among the rural women of reproductive age group. CONCLUSION: It is hoped that empowerment of FCHVs will bring sustainable change in their performance and will consequently produce notable improvement in the health of women and children in particular and in the community in general. PMID- 14741881 TI - General practitioners' perceptions of continuing medical education's role in changing behaviour. AB - CONTEXT: The effectiveness of moving to compulsory, prescriptive continuing medical education (CME) for New Zealand general practitioners (GPs) is questioned. Motivational interviewing theory suggests that a series of interventions gradually increase awareness of the need to change until change is finally actioned. This study aimed to explore GPs' views on their need for CME, experiences regarding its provision and perceptions on the effect of CME in changing their clinical behaviour. METHOD: Qualitative study using semi structured interviews of 24 GPs from Auckland and North Island rural areas assessing their CME experiences and preferences. FINDINGS: All participants acknowledged that CME is a life-long process essential for GPs. Changing behaviour is generally seen as an incremental, evolutionary process with reinforcement of knowledge from different sources. Single events were perceived to effect change rarely. These were often high-impact, either punitive or incentive-based. GPs have a myriad of CME sources including reading, the internet, specialist letters, conversations with colleagues, quality assurance feedback, as well as traditional meetings. Credit-based quota requirements received mixed opinions but mostly were perceived negatively, discouraging needs based approaches to learning. GPs' greatest barrier to obtaining CME is time. DISCUSSION: GPs perform poorly in assessing their specific learning needs. Their behaviour change is likely to be incremental. Therefore multi-faceted interventions and reinforcement from different sources are likely to be most effective in changing clinical practice. Understanding this is important for CME providers, GP Colleges and funders. Narrow, credit-based approaches to CME may discourage time-strapped GPs obtaining motivation to change from exposure to a wide variety of CME sources. PMID- 14741882 TI - Teaching children about health, part II: the effect of an academic-community partnership on medical students' communication skills. AB - CONTEXT: Partners in Health Education (PHE) is a Dartmouth Medical School elective that pairs first- and second-year medical students with local elementary school classroom teachers to provide health messages to students. The primary aim of the programme is to help medical students improve their communication skills through teaching children about health. Secondary aims are to teach children about health and the prevention of injury and disease and to support community teachers in their health promotion efforts. This report contains the results of the assessment of the programme's impact on the medical students. METHODS: Sixteen first- and second-year medical students comprised the participants for the study. Students were assessed during their first and fourth teaching experiences using a variety of evaluation measures, including student surveys of expectations and perceptions; ratings of performance in the classroom by students, teachers, and classroom observers; coded videotapes of classroom teaching sessions; and performance on a measure of physician-patient communication skills. FINDINGS: Over the course of the teaching experience, medical students' teaching and communication skills increased on a number of measures. DISCUSSION: Programmes such as PHE can provide true service-learning experiences in which all participants benefit. Medical students can learn how to communicate about health, a set of skills they will need to become effective physicians. School children can learn about health, so they are empowered to take charge of their health and to make healthy choices. PMID- 14741883 TI - Improving primary orthopaedic and trauma care in Nepal. AB - PURPOSE: To evaluate the efficacy of training programmes for the village health practitioners aimed at improving primary orthopaedic and trauma care in our rural setting. MATERIALS & METHODS: Six year prospective study of training workshops in local health institutions was carried out using locally available manpower and materials. RESULTS: After 6 years the evaluation of this programme shows a significantly improved (p < 0.05) knowledge base and working skills after completion of training workshop. SUMMARY: To avoid unnecessary complications, effective primary health care provided by an appropriately trained person should be available at the peripheral level. This training programme seems successful in achieving this goal and hence needs adoption for wider use in Nepal. PMID- 14741884 TI - Students' perceptions towards a family medicine attachment experience. AB - OBJECTIVE: To explore the students' perceptions about their experience in a family medicine (FM) preceptorship in order to provide a sound basis for offering guidance to family medicine undergraduate education. METHODS: During one full academic year at King Saud University, College of Medicine, Riyadh, Saudi Arabia, a self administered questionnaire was distributed to all students (n = 177) at the end of each 6 weeks family practice attachment course. Students were asked to rate their attachment by responding to a five-points Likert type scale questionnaire and other Yes/No questions reflecting different important points in teaching and training. RESULTS: The teaching quality and the ability of the General Practitioners (GPs) preceptors to relate to students are rated very highly. Despite that, the majority of students (59.3%) feel their aims were not met during the attachment. Students indicate that there should be more teaching of practical procedure skills, more time allocated for discussion and a greater student involvement in the consultation. CONCLUSION: There is much to be retained in FM preceptorships that involves the caring and communication aspects of learning patient care. The study showed that practical procedure skills are desirable features of a preceptorship programme and that an emphasis on doing vs. observing is preferred by students. Some conditions designed to improve preceptorships are outlined and basic practicalities of adding a preceptorship to a practice are considered. PMID- 14741885 TI - Setting up an innovative masters course in interprofessional health and welfare studies. PMID- 14741886 TI - Serum hepatitis: are surgeons and obstetricians putting themselves at risk? PMID- 14741887 TI - An interview of Professor Stephen Abrahamson. PMID- 14741888 TI - An interview of Albert Mutai. PMID- 14741889 TI - An interview of Elina Mantere. PMID- 14741891 TI - Position paper on community-based education for health professionals. PMID- 14741896 TI - Making community-based education programs sustainable? PMID- 14741898 TI - 'A touch of spice' - general practitioner registrars' views about cross-cultural communication. PMID- 14741899 TI - Using a modified course experience questionnaire (CEQ) to evaluate the innovative teaching of medical communication skills. AB - SETTING: At the Faculty of Medical Sciences, The University of the West Indies, St Augustine first-year students take two courses in health communication. In the Centre for Medical Sciences Education students completed a course experience questionnaire. This instrument is potentially useful in evaluating innovative programmes and securing support for their development. CONTEXT: The Faculty of Medical Sciences, The University of the West Indies, St Augustine was the first of three campuses to introduce compulsory health communication courses in 1995. Using a modified form of the 25-item course experience questionnaire (CEQ25) (Broomfield & Bligh, 1998) normally employed in overall degree or course evaluation, this study developed a 30-item questionnaire (CEQ30) to test its applicability to the evaluation of medical communication skills teaching MEASURES: In 1997 - 1998 medical, dentistry, veterinary medicine and pharmacy students completed the original short form of the CEQ25 including five items specific to medical communication skills teaching. Students used a five-point Likert scale ranging from one, indicating that they strongly disagreed with the statement, to a score of five, reporting that they strongly agreed with the statement. Principal Components Analysis with Varimax rotation analysed the scale structure of the evaluation tool. RESULTS: The principal components factor analysis of responses (n = 165) broadly confirmed five of Broomfield and Bligh's six factors, but identified a sixth factor in their original instrument (appropriate assessment) that split into two, and a seventh factor, use of available materials and resources. CONCLUSION: The modified CEQ30 is a reliable instrument with which to evaluate a course in medical communication skills, and encourage reflection on teaching and course design. Its use is applicable to medical courses during overall curriculum change and innovation in a medical school. PMID- 14741900 TI - Service learning in health care higher education: risk or not to risk. AB - The explosion of service learning as a teaching methodology in higher education has generated increased awareness of the potential for liability related to student experiences with service partnerships all over the world. At present, there are few instances of legal problems occurring using service learning with students, but risk management not risk avoidance may be the best form of preventing legal problems from occurring. Reviewing areas of potential liability with the service partner, the student, faculty and college institution for possible risks may reduce future problems. Strategies for dealing with potential identified risks are discussed in this article including identified agency risks, the capacities of the students, employment status, liability insurance, and agency incorporation. Reducing the risk with students includes evaluating the harm to and by students, assessing the student population, prerequisite courses, assumptions, developmental age of the student, good communication and use of an honor Code. The higher education institutions and faculty may reduce risk by instituting student contracts, maintaining student privacy, and using waivers. Good risk management through preparation of all involved in service learning projects is the key to success. PMID- 14741901 TI - Establishing a women's health curriculum using the Delphi Method. AB - CONTEXT: A Women's Health Initiative Task Force was formed with the collaboration of five primary care medical specialties at the University of Texas Medical Branch (UTMB) at Galveston to address the education of medical students about important issues related to women's health. OBJECTIVE: To identify and prioritize key concepts in issues related to women's health Methods: Literature review and three generational Delphi Method. RESULTS: The content was identified for development of a curriculum in Women's Health for medical students and resident physicians. Objectives, tasks, skills, learning opportunities, and learning materials were defined, and the curriculum is being designed to produce, within all specialties, parallel and consistent opportunities to educate students about issues important for women's health. CONCLUSION: The rankings of topics gathered though the Delphi Method will be used to develop a curriculum in Women's Health Issues that defines objectives, tasks, skills, learning opportunities, and learning materials for the purpose of educating UTMB medical students and resident physicians about the health needs of women. PMID- 14741902 TI - Clinical experience of medical students in a developing country. AB - OBJECTIVE: This paper compares the clinical experience in acute conditions of the undergraduate students of a medical school from a developing country (Malaysia) with those from a developed country (UK). METHODS: This study was conducted at the School of Medical Sciences, Universiti Sains Malaysia (USM). Through questionnaire survey enquiry was made about 27 acute medical conditions (i.e. conditions related to internal medicine, paediatrics, and psychiatry), 15 acute surgical conditions (i.e. conditions related to general surgery, orthopaedics, ophthalmology, otorhinolaryngology, gynaecology and obstetrics), 15 surgical operations and 26 practical procedures. The results obtained were compared with published data from the UK. RESULTS: Acute medical conditions were seen by higher number of the USM students but with less frequency than the British students. The USM students saw practical procedures more frequently than the British students did, but almost an equal number performed these procedures independently. The British students attended surgical operations more frequently than the USM students did. CONCLUSION: Given the limitations of comparison (epidemiological, cultural and geographical differences, conventional curriculum (in the British medical schools) vs. problem based learning curriculum (in the Malaysian medical school)) the overall clinical experience of the medical students in the USM and the UK was comparable. The USM students had more opportunities to observe cases and procedures but "hands on" experience was similar to that of the British students. PMID- 14741903 TI - Graduate health professions education: an interdisciplinary university - community partnership model 1996 - 2001. AB - INTRODUCTION: In 1996, East Tennessee State University (ETSU) reinforced its historical commitment to multidisciplinary community engagement by developing a graduate level community partnerships program in the Division of Health Sciences. While the university's earlier health partnership efforts relied primarily on curricular innovation, the approach to graduate health professions education was to seed a series of curricular enhancements and interdisciplinary, community based learning experiences and service into traditional curricula. This paper presents the experience of one school in crafting a regional network that became the basis of a division-wide graduate level teaching and learning initiative. INNOVATIONS AND EVALUATION: Carefully selected planning and implementation techniques enabled multidisciplinary practitioners and community members from across a 20-county region to participate with university faculty in training ETSU learners in community-based medical care. By year four of the project, curricular "enhancements" were institutionalized in over five departments across the Division and engaged 1160 medical residents and graduate learners in a give - get model of health education. Programme evaluation methodology was collaboratively defined and documentation of programme effort and outcomes regularly reported and strategically reviewed. CONCLUSIONS: Programme evaluation demonstrates mutual benefit to community and university. Faculty involvement in programme activity increased fourfold and community involvement in training of health professions graduate learners increased threefold by year four. Educational innovations were adopted into traditional curricula, thousands of hours of clinical services were provided to underserved communities and the university-community team forged by network links continues to promote multidisciplinary interests through joint public policy endeavors. PMID- 14741904 TI - Student generated learning objectives: extent of congruence with faculty set objectives and factors influencing their generation. AB - CONTEXT: A problem-based learning strategy is used for curriculum planning and implementation at the Arabian Gulf University, Bahrain. Problems are constructed in a way that faculty-set objectives are expected to be identified by students during tutorials. Students in small groups, along with a tutor functioning as a facilitator, identify learning issues and define their learning objectives. PURPOSE: We compared objectives identified by student groups with faculty-set objectives to determine extent of congruence, and identified factors that influenced students' ability at identifying faculty-set objectives. METHOD: Male and female students were segregated and randomly grouped. A faculty tutor was allocated for each group. This study was based on 13 problems given to entry level medical students. Pooled objectives of these problems were classified into four categories: structural, functional, clinical and psychosocial. Univariate analysis of variance was used for comparison, and a p > 0.05 was considered significant. RESULTS: The mean of overall objectives generated by the students was 54.2%, for each problem. Students identified psychosocial learning objectives more readily than structural ones. Female students identified more psychosocial objectives, whereas male students identified more of structural objectives. Tutor characteristics such as medical/non-medical background, and the years of teaching were correlated with categories of learning issues identified. CONCLUSION: Students identify part of the faculty-set learning objectives during tutorials with a faculty tutor acting as a facilitator. Students' gender influences types of learning issues identified. Content expertise of tutors does not influence identification of learning needs by students. PMID- 14741905 TI - Problem-based learning in online vs. face-to-face environments. AB - This study compared outcomes of problem-based learning between synchronous online groups and face-to-face tutorial groups. Specifically, the study compared learning outcomes, time-on-task and learning issue generation. METHODS AND PROCEDURES: A post-test only control group design was used to investigate the effects of learning conditions on learning outcomes and processes. The experimental learning condition was defined as computer-mediated problem-based learning (CMPBL) and the control condition was traditional problem based learning in face-to-face groups (TPBL). The learning process consisted of four elements: an initial tutorial, a period of self-directed learning, a second tutorial and a laboratory session. During the initial tutorial students generated learning issues that they submitted to the research assistant. In the self-directed learning phase, students researched their learning issues and returned for the second tutorial with their findings. Students in the CMPBL groups interacted with the resource person electronically via email, chat room or bulletin board. At the second tutorial, groups shared information related to their learning issues and completed their products for the problem. RESULTS: There was no difference in learning outcomes between groups. The CMPBL group spent significantly more time on learning than the TPBL group. There was no overall difference between groups on generation of learning issues; however, there was a significant relationship between number of learning issues generated and higher score on the examination regardless of tutorial medium. PMID- 14741906 TI - From quality assurance to total quality management: how can quality assurance result in continuous improvement in health professions education? AB - CONTEXT: Quality assurance is a hot topic in many institutes for higher education. Quality assurance is a cyclic process, consisting of three steps: measuring, judging and improving. Many institutes in higher education collect data about the quality of their educational programme on a regular basis and with proper frequency, but these evaluative data are not always used to improve the quality of the programme. PURPOSE: The aim of this article is to demonstrate which conditions need to be fulfilled to ensure that quality assurance is a cyclic process resulting in continuous improvement. CONCLUSIONS: Quality assurance can only be successful, i.e. result in continuous improvement, if three conditions are met: the evaluation activities are carried out in (1). a systematic and (2). structural fashion and (3). are integrated in the organization's regular work patterns. Parts of the system for quality assurance at the Maastricht Medical School are presented to demonstrate how the three conditions can be realized in practice. PMID- 14741907 TI - Community-based education programme of Dokuz Eylul School of Medicine. PMID- 14741908 TI - Educating nurses to build a care-with-confidence team. PMID- 14741909 TI - The world health report 2002 - reducing risks, promoting healthy life. PMID- 14741910 TI - An interview of Benjamin Stockins. PMID- 14741911 TI - An interview of Ryan Camado Guinaran. PMID- 14741913 TI - Reflections on the network: TUFH's conference in Eldoret, Kenya. PMID- 14741919 TI - A new socially responsible medical school for regional Australia. AB - BACKGROUND: Northern Australia is a geographically large region that has too few medical graduates and is a long way from Australia's metropolitan medical schools. This paper describes the establishment of a new medical school to serve these needs. METHODS: The paper describes the steps in development and presents an overview of the innovative educational approach, based on best available educational evidence, that aims to produce a medical workforce that understand the health care needs of the region and want to practice there. CONCLUSION: The result is an innovative, highly integrated programme with different aspirations and a different student profile from other Australian medical schools. PMID- 14741918 TI - Evaluation of senior citizens' satisfaction in primary health centres as assessment of the academic model. AB - CONTEXT: Satisfaction is nowadays a valid measure of quality of care. Senior citizens are increasing in Chile and their treatment in primary care clinics, as well as the education of new professionals must take into consideration their special characteristics. GOAL: Our intention was to investigate the degree of satisfaction senior citizens had with their health service and to identify those aspects able to be modified at the Medical Faculty in order to improve education of health professionals. APPROACH: Three hundred elderly attending Primary Care Outpatient Clinics of the city of Temuco, Chile, were interviewed using a satisfaction questionnaire developed by the Medical Outcomes Study carried out in USA. FINDINGS: Senior citizens experience a high level on dissatisfaction with the health care they receive at the primary level. Doctors and nurses had a high qualification in relation to their technical skills, but a lower score if considering the education and information they give to patients. These results shall be used in order to modify attention to senior citizens as well as to include problems related to this group in the curricular reform plan and to improve attitudes of nursing and medical students. PMID- 14741920 TI - Changing practice in residential aged care using participatory methods. AB - BACKGROUND: Residential aged care staff play a significant role in the day-to-day lives of residents yet are faced with many barriers to providing care that promotes resident wellbeing. APPROACH: Action research is a useful approach for clarifying issues, identifying education and training needs, and identifying, and in some cases overcoming, organizational barriers to change. The Well for Life project aimed to enhance the social and physical health and well being of residents of aged care settings by empowering the staff of facilities to make change. The project had a particular focus on nutrition and physical activity. This paper reports on the action research group process undertaken during Phase I of the Well for Life project. Five residential aged care settings participated in the action research process facilitated by project staff independent of the facilities. The action plan and outcomes from one of these settings is used to illustrate the process and outcomes. FINDINGS: The main findings of the project indicate that using a process that encourages staff involvement in identification of issues and actions can facilitate change in the practice of resident care. The action research groups identified specific gaps in knowledge and skill leading to targeted education that addressed areas of need. The importance of presenting information and learning opportunities for staff in a variety of formats was also recognized, as was the importance of organizational context, management support and empowerment of staff to make change. PMID- 14741921 TI - Teaching psychiatry in poor countries: priorities and needs. A description of how mental health is taught to medical students in Malawi, Central Africa. AB - CONTEXT: In developing countries poor standards of clinical service may be an obstacle to medical education. The paper outlines the inadequate mental health service in Malawi, Central Africa, which as well as failing patients obstructs the training of health workers. AIMS AND OBJECTIVES: A new mental health curriculum at Malawi's medical school is described. The notion of psychiatry as a medical specialty is abandoned and the focus moved to psychological and psychiatric factors as they present in general clinical settings and primary care. Students are encouraged to consider how they may address mental health issues through the many and varied roles which doctors in resource poor countries must fulfil (administrator, trainer, primary health care doctor, hospital physician). RELEVANCE: This training strategy accords with a shift of attention among health service planners world-wide from specialist services towards primary care, and may generalize to other settings in which specialist clinical services are poorly developed. PMID- 14741922 TI - A memory of an aesthetic experience transferred to clinical practice. AB - PURPOSE: To examine the usefulness of writing about a memory of an aesthetic experience, and then transfer the aesthetic experience to a health care situation. METHODS: The study was accomplished at two university colleges of health sciences in Sweden. It started with student nurses (N=291) writing about a memory of an aesthetic experience. Then they transferred the aesthetic experience to a purposeful clinical practice. FINDINGS: The results showed that each student could report on a positive memory of an aesthetic experience. Embedded in each story was an aesthetic experience that was meaningful to the student. Domains of memory most frequently reported were music, work of art and nature. Themes derived from the aesthetic memory were happiness and awareness. The awareness theme comprized the value of aesthetic experiences for the patients, and for student nurses. CONCLUSION: The process of writing about a memory of an aesthetic experience provided an alternative model for nursing education that could improve patient care. PMID- 14741923 TI - What medical students value in a population health tutor: characteristics for consideration in staff recruitment and development. AB - BACKGROUND: Recent moves to integrate aspects of population health into medical curricula have created new staff development challenges for many institutions. Small group teaching in population health is a relatively new initiative, often requiring recruitment of additional staff and considerable training. This analysis identifies the tutor characteristics rated most positively by medical students in a small-group course in population health and discusses their implication for staff recruitment and development. METHOD: Retrospective evaluation of tutors by students using a self-administered questionnaire. Overall tutor rating was analysed against various tutor characteristics, using univariate logistic regression methods. Optional qualitative comments were summarized by thematic methods and triangulated with findings from the quantitative analysis. RESULTS: Creating a supportive group climate was the tutor attribute most positively evaluated by students (OR=9.62, 95%CI 4.46-20.83). Perceived interest in teaching (OR=8.93, 95%CI 3.83-20.83) and the ability to give useful feedback (OR=8.40, 95%CI 4.07-17.54) were also highly rated by students as valuable qualities in their tutors. Qualitative analysis highlighted the importance of informed comment, good knowledge and expert input from tutors. CONCLUSION: Whilst a degree of content expertise in population health was desirable in a tutor, its value was secondary to good facilitation skills and an enthusiasm for teaching when student evaluation was considered. Faculties implementing small-group methods of teaching population health should consider facilitation skills and interest in teaching as priorities when recruiting and training staff. As is the case for self-directed student-led learning in basic and clinical sciences, these appear to be more influential than content expertise, from the students' perspective. PMID- 14741924 TI - Format of cases affects learning outcomes in first year medical students. AB - CONTEXT: Longitudinal problem-based learning (PBL) tutorials are practiced at the Tokyo Women's Medical University. First year medical school students - most of whom are high school graduates with no medical background - often encounter difficulty identifying problems while solving PBL cases in basic science. The format of PBL case presentation may affect learning. OBJECTIVES: This study compares the learning outcomes of two cohorts of first year students who learned basic human biology through PBL cases presented in clinical vs. non-clinical formats. METHODS: All first year students in 1995 and 2000 undertook PBL tutorials. The 1995 case was presented in a non-clinical format; the 2000 case was presented in a clinical format. Both cases had five identical pre-set learning objectives in basic science. By examining all written materials generated during the tutorial sessions, learning outcomes were categorized and the accomplishment of preset objectives was analysed. FINDINGS: In 2000, the number of learning outcomes for clinical medicine was more than double compared to 1995, whereas the numbers of total and basic science learning outcomes were not significantly different. The number of preset objectives accomplished by the students was significantly higher in 2000. Thus, PBL case format affected the learning outcomes, enabling these first year students to achieve basic science objectives, while enhancing their interest in the clinical aspects of human biology. CONCLUSION: Learning outcomes in first year medical students may be enhanced when PBL cases designed to learn basic science contain relevant clinical elements. PMID- 14741925 TI - The role of reflective practice in pharmacy. AB - CONTEXT: There has been a considerable paradigm shift from a product-centred focus towards a patient-centered focus in pharmacy. Pharmaceutical care practice (Cipolle et al., 1998) can be seen as the latest proposal to transform the profession as mandated by most major professional associations in pharmacy. Pharmaceutical care is an innovative way of practicing pharmacy that has the potential to make drug-therapy safer, more effective, and more convenient for the patient. Even though pharmacists' time spent on customer communication has increased over time (Savage, 1999), this alone will not be sufficient to bring about a paradigm shift in view of what the pharmacist actually does to provide quality patient care. It appears equally important to reflect on how pharmacists practice pharmacy. METHODS/OBJECTIVES: It is the aim of this discussion paper to argue for the importance of linking education and practice as a core part of the teaching of pharmaceutical care. CONCLUSION: In order for pharmacists to partake in truly interdisciplinary health care teams and the profession of pharmacy to demonstrate its unique and indispensable contribution to quality health care, pharmacy curricula would teach toward reflective practice, be problem-based, be positioned in collaborative teams, and have an outside (authentic) focus. PMID- 14741926 TI - An integrated approach through health, education and community development for the developing countries: the Baqai model. PMID- 14741927 TI - Regression toward the mean: neither good nor evil, just unplanned. PMID- 14741928 TI - Interview of Fernando Mora-Carrasco. Interview by Jan van Dalen. PMID- 14741929 TI - An interview with Lupe Ramos, promotora in Fabens, El Paso County, Texas, USA. Interview by Judith V. Sayad. PMID- 14741930 TI - Interview of Jervas Victor Dimo, a Sudanese medical student at Moi University in Eldoret, Kenya. Interview by Jan van Dalen. PMID- 14741936 TI - Why is it taking so long? PMID- 14741937 TI - A longitudinal, patient-centred, integrated curriculum: facilitating community based education in a rural clinical school. AB - The Greater Murray Clinical School (GMCS) was founded with two main aims in mind one, to provide a community-based learning environment offering diversified clinical educational experiences, and two, by doing so, to help address the doctor shortage for Australians living in rural and remote areas. The GMCS is a community-orientated and community-based clinical school, which has replaced the typical discipline-based curriculum with a longitudinal, patient-centred one. Students are attached to patients--called "the longitudinal patient"--whom they follow through all stages of their care. They share with patients their experience of illness and disease, their varying care needs, and how these are addressed by different service providers. The philosophy of the course, its implementation and our initial experiences are described. PMID- 14741932 TI - Position paper on problem-based learning. PMID- 14741938 TI - Introducing psychosocial and psychiatric concepts to first year medical students using an integrated, biopsychosocial framework. AB - OBJECTIVE: The main objective of the problem-based teaching unit reported in this paper was to introduce psychosocial and psychiatric concepts to first year medical students using an integrated approach. METHOD: A total of 131 undergraduate students studied a case of delirium. Students were encouraged to understand the problem from a number of perspectives and approaches. In particular, the patient's view was emphasised. This was partially achieved by employing a standardised patient, who answered students' questions about what it was like to be unwell and hospitalised. Both quantitative and qualitative evaluations of the teaching project were conducted. RESULTS: Overall, the teaching project was well received. However, as an introduction to a complex and unfamiliar area, students were concerned that material was difficult to grasp. CONCLUSIONS: As a foundation for future teaching in psychiatry, the case and our methods appear appropriate. However, this initial teaching should be reinforced and expanded upon in all years of the medical curriculum. PMID- 14741939 TI - Intern prescribing decisions: few and far between. AB - OBJECTIVE: To examine the scope of intern prescribing practices by determining: the proportion of prescriptions that interns chart compared with other medical staff; the proportion of intern-charted prescriptions for which interns are sole decision-makers; whether or not intern-initiated prescribing varies with respect to the specialty to which they are attached, the shifts they are working and the types of charts they are using; the types of clinical conditions for which interns initiate prescribing decisions; and the drug classes that interns use for their self-initiated prescribing. DESIGN: Prospective study of a random sample of intern-charted prescriptions. SETTING: Two teaching hospitals of the Hunter Area Health Service, Newcastle, Australia. The study was conducted from the fifth to the eighth month of the intern training year. MAIN OUTCOME MEASURES: The proportion of prescriptions charted by interns that resulted from their own decision-making, the circumstances relating to this, clinical conditions for which they prescribe and drugs prescribed. RESULTS: A total of 17,895 prescriptions were examined--3437 (19%) were intern-charted. Interns reported they were the sole decision-makers for 19% (95% CI: 14 -24%) of prescriptions they had charted. Interns were more likely to initiate decisions in accident and emergency (OR=7.5, 95% CI: 2.2- 25.2) and obstetric and gynaecology (OR=2.3, 95% CI: 1.6 -3.2) rotations than in medicine and were more likely to initiate decisions on night (OR=7.3, 95% CI: 3.4- 15.5) and weekend (OR=1.7, 95% CI: 1.0 3.2) shifts than during the day. They were also more likely to prescribe on the "as required" (OR=36.6, 95% CI: 20.6-65.0), "statim" (OR=26.1, 95% CI: 17.0- 40.1) and "intravenous" (OR=7.2, 95% CI: 4.3-12.3) charts compared with "regular" charts. A total of 52% of intern-initiated prescriptions were for symptom relief; pain, insomnia and nausea; and 75% of the drugs for which interns made independent decisions were analgesics, antithrombotic agents, psycholeptics, antispasmodics, laxatives and anti-asthmatic agents. CONCLUSION: Interns have a limited role with respect to independent prescribing and take the sole responsibility for only one-fifth of the prescriptions they chart. This limited, albeit safe, approach to prescribing may impact significantly on an intern's opportunity to acquire the skills necessary to become an independent, rational prescriber. PMID- 14741940 TI - Inside PBL groups: observation, confirmations and challenges. AB - This article is based on a study of nine problem-based learning (PBL) groups with eight medical students and one tutor in each. The groups were observed at their two-weekly meetings for three weeks. The observations were recorded by video and written reports. Three illustrative examples of these observations are presented and discussed. The purpose of the study was to understand how processes in PBL groups correspond with the theory-based intentions for this type of learning and teaching and to discuss the theoretical foundations for problem-based learning. The observations confirm the expectation that groups in the observed program would follow a systematic stepwise progression. The groups were also found to become more flexible and self-directed in their use of PBL as they became more familiar with the approach, provided that the tutor did not dominate the interactions. It is argued that processes in problem-based learning should be understood not only in the light of theories focusing on individual learning and knowledge construction, but also from other theoretical perspectives. Socio cultural perspectives with constructs such as collaborative learning, apprenticeship and modelling shed new light on what happens inside PBL groups in ways that will enhance learning and tutoring. PMID- 14741941 TI - The Dutch system of external quality assessment: description and experiences. AB - CONTEXT: Generally speaking, quality assurance in Dutch higher education consists of four parts: a self-evaluation, written by the faculty, a review committee, consisting of experts, the site visit, and a public report of the assessment outcomes. TASK OF THE REVIEW COMMITTEE: The organization, in which all universities are united, appoints a review committee for each educational program. Most educational programs are taught at different universities. Consequently, the review committee visits several universities. The task of the review committee is: to assess the overall quality of these educational programs in The Netherlands; and to assess the quality of each school separately. METHOD: The members of the review committee start their activities by formulating the evaluation criteria (standards) for the content of an educational program and for the educational process. This ends with a paper that will be used as a reference for the assessment. Then the review committee studies the self-evaluation reports. The faculties write these reports along established guidelines. In cases where the self-evaluation is not completely clear, or not sufficient, the committee formulates questions and asks for additional information. Then the committee visits the faculties. These visits take usually two days each. During these visits, the committee gets information from the dean of the faculty, the program committee, teachers, students, and from other employees of the faculty. In addition, the members of the committee inspect the facilities of the faculty, e.g. the library. RESULT: Directly after the visit, the chair of the committee gives, orally, a first impression of the findings. After visiting all faculties, the committee starts their deliberations, which result in a report of the assessment of the overall quality of the educational programs. The committee also assesses the quality of each faculty separately, often ending with recommendations. CONCLUSION: The Dutch system of quality assurance in higher education is much copied by other countries. The essence is the self-evaluation on one hand, and the site visit on the other. Committees try to act as consultants, and not as executioners. Therefore, the way the members of the committee play their role is very important. PMID- 14741942 TI - The tropical triangle: a health education alliance for the Southwest Pacific. AB - INTRODUCTION: Few collaborations between universities in developed countries and medical schools in developing countries have been described in detail in the medical literature. We describe a collaboration between three medical schools, one in a developed country and two in developing countries based on shared challenges and missions. THE ALLIANCE: James Cook University School of Medicine (JCU), Fiji School of Medicine (FSM) and University of Papua New Guinea School of Medicine (UPNG) are all located within the tropical regions of the Southwest Pacific. All schools serve vast geographical areas where much of the population lives in rural and remote communities with limited access to medical care. JCU's first class started in 2000 as the only complete medical school in Tropical Australia and was founded with a mission to meet the health care needs of rural, remote and underserved populations in the region. FSM educates medical students from most English-speaking developing Pacific Island Nations. UPNG serves a predominantly rural developing nation where infectious diseases and other diseases of rural developing nations predominate. Based on their common challenges and goals, the three schools established an informal collaborative relationship called "the Tropical Triangle" in the late 1990s. OBJECTIVES, ACTIVITIES AND CHALLENGES: These very different institutions are committed to an effective partnership based on mutual understanding and knowledge of each other's day-to-day challenges. Faculty development seminars on medical education have already been carried out in Fiji by JCU staff. JCU has also offered several PhD scholarships to FSM staff, three of which are in the process of being taken up. JCU has offered to make available its online teaching resources to FSM and UPNG. Student exchanges are planned, and FSM and UPNG have exchanged examiners on several occasions for the MBBS and postgraduate programs. The possibilities for collaborative research on regional problems are being explored. CONCLUSION: The Tropical Triangle Alliance is a special and perhaps unique alliance based on equality and shared challenges. The benefits from this alliance flow both ways. The alliance may serve as a model for other collaborations between health education institutions in developed and developing countries. PMID- 14741943 TI - Service-learning in healthy aging for medical students and family medicine residents. AB - INTRODUCTION: Community-based educational opportunities can diversify and strengthen traditional clinical education. With growing diversity of patient populations and increasing life expectancy, it is imperative that medical students and residents prepare for practice within this context. The Center for Healthy Communities in the Department of Family and Community Medicine at the Medical College of Wisconsin, USA developed a community-based, service-learning program in healthy aging to address this need. OBJECTIVES: The goals of the Chat and Chew program are to: generate medical student/ resident awareness of community health, aging, and diversity; train medical students/ residents to present health information to older, minority community members; encourage medical students/residents to view community members as "teachers" as well as patients; and provide needed health information and socialization opportunities to elderly public housing tenants. IMPLICATIONS: Medical students and residents gain the opportunity to interact with community members about the health issues that concern them. They also benefit from seeing community members in their real life context and learning about their health-related experiences. The housing tenants help shape how future patients will be understood and treated by the physicians who participated in the service-learning program. The purpose of this article is to: (1) provide an overview of service-learning and the Chat and Chew program, including reflection components; and (2) discuss how this program has become an integral part of the family medicine residency curriculum. PMID- 14741944 TI - The ambiguous and bewitching power of knowledge, skills and attitudes leads to confusing statements of learning objectives. AB - The words "knowledge", "skills" and "attitudes" are given different meanings by health personnel when discussing educational issues. Ambiguity is known as a handicap to efficient communication. In the design of a curriculum the quality of the definition of learning objectives plays a fundamental role. If learning objectives lack clarity, learners and teachers will face operational difficulties. As Robert Mager said, "If you are not certain of where you are going you may very well end up somewhere else and not even know it". Knowledge is not only memory of facts but what you do with it. The complexity of human behaviour should not be underestimated. This is why educational objectives need active non-ambiguous verbs in order to achieve better communication between teachers and learners and to assess that complexity. This is why I suggest using the expression intellectual skill (or competence) as meaning "a rational decision or act". Sensomotor skill (or competence) would replace "skills" as presently used and cover only "acts which require a neuromuscular coordination". Interpersonal communication skill (or competence) would replace "attitude(s)" and be limited to "verbal and non-verbal relation between persons". As the level of validity of assessment of learners' competencies is linked to the clarity of learning objectives, it is hoped that the above suggestions will raise the overall level of validity of the evaluation system. This is why it is important that everybody understands, in the same manner, the meaning of a learning objective. It will help learners to focus their learning efforts on the right target. It will help teachers to ensure the relevance to health needs of their teaching and the validity of assessment instruments. In both cases it will be beneficial to the health of the population. PMID- 14741945 TI - Who's my doctor today? AB - Clinical practice is not always user-friendly. Specialisation fragments patient care across different investigations and modes of management. Increasing hospital throughput, especially by day surgery, diminishes the time available for students and other health professionals in the team to interact with the patient and verify the appropriateness of the care plan. Patients are at a serious disadvantage in ensuring that their concerns are understood, and in negotiation of which management plan would optimise the outcomes they seek. PMID- 14741946 TI - Informational, interpersonal, and intrapersonal communication in a family practice resident support group. AB - CONTEXT: This prospective study compared group process literature to topics utilized in a year-long family practice resident support group. OBJECTIVES: The basic hypothesis was that resident support groups function in ways that are similar to other groups to the extent that the literature on group process could contribute to our understanding of support groups for residents. METHOD: Ten possible discussion topics were grouped along a continuum from informational to interpersonal to intrapersonal. ANALYSIS: Data were subjected to three repeated measures of analysis of variance (ANOVA). FINDINGS: Results reflected that informational topics did not decrease, interpersonal topics did increase significantly, yet intrapersonal topics did not reflect any significant changes over the course of study. DISCUSSION AND CONCLUSIONS: The preliminary findings indicated the research literature on group process may have application to resident support groups. When more formalized groups like Balint groups are not available, support groups may offer a forum to facilitate the interpersonal and intrapersonal discussions and communications of residents. Recommendations derived from the support group experience and the research are given. PMID- 14741947 TI - Teaching and learning about bioscience ethics with undergraduates. AB - Bioscience ethics acts as a practical interface between science and bioethics. It links scientific endeavour and its application into adaptive forms of bioethical consensus. Its major elements are increased understanding of biological systems, responsible use of technology, and curtailment of ethnocentric debates in tune with new scientific insights. This paper briefly describes the students' learning experience gained from the vacation unit BIOL 240, Introduction to Bioscience Ethics, as taught in biology, Macquarie University. On the basis of our evidence students were overwhelmingly positive about their learning because the unit assisted them to better face dilemmas that arise from the application of science and technology. The structure also provided active engagement with the subject matter and preferred learning environments that supported and contested their understanding of concepts relevant to bioscience and bioethics. PMID- 14741949 TI - Interactive lecture supported by multimedia presentation: a new teaching tool for faculties with crowded classes and limited budgets. PMID- 14741948 TI - A tale of two exposures: a comparison of two approaches to early clinical exposure. AB - PURPOSE: Early Clinical Exposure (ECE) programs are an increasingly widespread component of undergraduate medical education. Little systematic research exists on the topic. This report compares the approach and cost/benefits of two parallel ECE programs at the Sackler School of Medicine, Tel Aviv University. METHOD: The two ECE programs were compared using student questionnaires, faculty questionnaires, focus groups and participant-observations. RESULTS: ECE for American medical students in Israel was based upon a semi-structured mentoring relationship with a hospital-based specialist, in which students were able to practice interviewing skills. ECE for Israeli students emphasized structured exposure to a wide variety of primary care settings and informal patient contact. Both ECEs were supplemented by campus-based small group discussion. Students, preceptors and group leaders in both ECEs all reported very high levels of satisfaction and perceived impact on medical training. CONCLUSION: There appears to be no "best" way to conduct ECE. The ECE should be tailored to the specific needs and goals of the student population. Further research and international comparison is needed. PMID- 14741950 TI - An interview with Gerard Majoor. Interview by Jan van Dalen. PMID- 14741951 TI - An interview of Tim Dodge. Interview by Jane Westberg. PMID- 14741958 TI - What does "community-oriented" mean anyway? Some thoughts on Zohair Nooman. PMID- 14741959 TI - Zohair Nooman: a personal memoir. PMID- 14741960 TI - Relationships: a new way to analyse community-based medical education? (Part one). AB - Relationships do matter! In fact, medicine cannot be learned without them, and community-based medical education (CBME) curricula that ignore them or take them for granted do so at their students' peril. As CBME is becoming more popular, there is a need to develop appropriate frameworks for describing quality in CBME to ensure that it remains a principle-driven, not format-driven, initiative. In this paper, I provide evidence for a simple model of four key relationships, the four Rs, in which the medical student must be immersed to facilitate high quality learning. These four Rs are the relationships between (1) clinicians and patients, (2) health service and university research, (3) government and community, and (4) personal principles and professional expectations. As a result of this synthesis of the current medical education literature, I propose that this model of clinical, social, institutional and interpersonal relationships is a valid framework for articulating the important principles in CBME, and that it describes why community-based medical education is such an attractive alternative. Relationships do matter! PMID- 14741961 TI - Integrity: the key to quality in community-based medical education? (Part two). AB - Relationships do matter! In fact, medicine cannot be learned without them, and community-based medical education (CBME) curricula that ignore them or take them for granted do so at their students peril! Could these assertions provide a key to quality in CBME curricula? In a previous paper, I provided evidence for a simple model of four key relationships, the four Rs, to act as a framework to analyse CBME. These four Rs are the relationships between (1) clinicians and patients, (2) health service and university research, (3) government and community, and (4) personal principles and professional expectations. In this paper I describe a key to discerning quality in this model. This key is integrity - that coalition of soundness and interdependence that enables the quality of the whole to be greater than the sum of the parts. I will also discuss the implications of this model for CBME with particular reference to reform of student selection, curricular goals, course structure, and assessment. Whilst further research based on this model is needed, as a result of this synthesis of the current medical education literature, I propose that it is its capacity to produce clinical, social, institutional and interpersonal integrity in its graduates that makes community-based medical education such an attractive alternative. Relationships do matter! PMID- 14741962 TI - Evaluation of an innovative approach to community-based medical undergraduate education in Nigeria. AB - CONTEXT: Although innovative, community-oriented and PHC-focused medical education programmes have been in operation in some medical schools in Nigeria for over a decade, they are yet to be comprehensively evaluated. OBJECTIVE: This study therefore aimed at evaluating some impacts of the programmes on medical education in the country. METHODS: The study was conducted in three innovative medical schools in South-Western Nigeria. Two traditional medical schools were selected as control. Questionnaires were used for the collection of data from random samples of 44 final year medical students in the innovative medical schools (SIMS) and 40 final year medical students in the traditional medical schools (STMS). Forty (40) medical graduates of the innovative medical schools (GIMS) and 33 graduates of the traditional medical schools (GTMS) also participated in the study. In addition, in-depth interviews of key stakeholders of the programmes and focus group discussions of selected members of the communities were conducted. FINDINGS: Findings revealed that the graduates of the innovative schools were better exposed to PHC education than those in the traditional schools. Their perceptions of the objectives of, and functions during, the PHC education were significantly different. Methods of learning during the programmes appear to be more experiential and inductive. Attitudes of members of rural communities were also favourable to the programmes. CONCLUSION: The innovative programmes appear to have impacted positively on medical education in the country. A major deficiency of the programmes is inadequacy of human and material resources for their effective functioning. PMID- 14741963 TI - The impact of a community-oriented medical school on mental health services. AB - This paper discusses the impact of the community-based medical school on mental health services. The Gezira mental health programme represents a collaborative work involving the university, the community and the government. It aims at achieving specified objectives: (1) to modify community concepts, attitudes and practices concerning mental health, (2) to ensure community involvement and participation, (3) to extend mental health services, (4) to train PHC staff, and (5) to encourage research. The programme was implemented in three phases: preparatory, implementation, and evaluation. In the evaluation of the impact of the programme on changing community attitudes, the training of staff, the extension of mental health services, and on research, qualitative assessment, through interviews, focus group discussion, supervision visits, and review of reports are used. There is an overall agreement that the programme helped in raising public awareness regarding the concept of mental health, the care of the mentally ill and community participation. Members of the health team who received training as part of the programme reported a better understanding of mental health problems and an improvement in their handling of the mentally disturbed patients. Teachers reported an increased awareness of mental health problems in school children and a better collaboration with those involved in the handling of such problems. Social workers and psychologists updated their knowledge and skills and were well prepared to participate in the programme. Members of the different sectors involved reported a better standard of collaboration regarding mental health activities. These findings indicate that this programme, by providing a new model for health services in this field, has induced a large policy change within the Sudan. The community-based activities at the FMUG have resulted in a major change in the delivery of mental health services in Gezira State. The programme has resulted in a major shift in mental health services being provided by central hospitals to PHC settings. In addition it has stimulated research, thereby providing much original information that will help in preparing for future plans. PMID- 14741964 TI - A description of a community-oriented cum PBL post graduate training course for health districts managers in Central Africa. AB - CIESPAC (Centre inter-etats d'Enseignement en SantePublique pour l'Afrique centrale), a sub-regional public health training institution, originally located in Brazzaville, was created with the vocation of providing Central African countries with qualified health services managers (the turbulent events that occurred in Brazzaville in June 1997 prompted the transfer of the institution to Yaoundein Cameroon). It offers several courses, the most recent of which culminates with a professional diploma in public health and targets mainly potential health district managers (DPSP--Diplome Professionnel en SantePublique). This paper reviews the first four-year experience of implementing the "community-oriented problem-based learning" (PBL) pedagogic approach in francophone Africa. About 70 health professionals (mainly MD and diploma nurses) were trained, using the PBL approach, within a period of three years. Practical field training activities involving the neighbouring urban communities of the institution as training sites were given a key place in the course and thus allowed trainees to perceptively appraise the priority health problems of a district. The most important thing trainees learnt during this course was how to learn. Some of them were also able to participate in some operational research. From this experience, it is clear that trainees are coached to learn solving problems on their own for the rest of their professional career. The neighbouring community of a training institution, when properly approached, can provide a very fertile teaching ground where trainees can acquire first hand practical experience in learning to collaborate with local communities. However, the PBL pedagogic approach requests a mutual understanding between trainees and trainers. As such there is a need for a careful selection of trainees and, even more importantly, for a critical mass of competent and motivated trainers. PMID- 14741966 TI - Developing an OSCE station to assess the ability of medical students to share information and decisions with patients: issues relating to interrater reliability and the use of simulated patients. AB - CONTEXT: Patient partnership is being promoted as an aid to compliance with treatment and to improve outcomes for patients. An integral part of this partnership is information-sharing between doctor and patient and negotiation of management decisions, together with an ability by the doctor to analyse critically treatment options. These skills are being taught to some extent in communication skills for medical students and assessments are being devised to test them. OBJECTIVES: To develop an OSCE station to assess medical students' skills in the application of evidence and the negotiation of treatment options with a simulated patient. To evaluate the station by tests of reliability and comparison of the marking of observing examiners and simulated patients. DESIGN: An OSCE station was devised using a case scenario in which there were varying opinions as to the treatment options. The marking schedule was designed to assess the students' ability to assess evidence and discuss this with patients. SETTING: University of Leeds. SUBJECTS: Undergraduate medical students. RESULTS: A total of 194 students undertook the station. Inter-rater reliability amongst examiners was poor for individual student marks but there was better agreement as to whether a student passed or failed. There was poor agreement between the observing examiners and the simulated patients for individual marks. The examiners thought this was a useful attempt to assess students' ability to give information but agreed that further work was needed on the marking schedule. CONCLUSIONS: More work is needed to refine the station to increase its reliability. However, the station is useful in highlighting to students the importance of sharing information and decision-making on treatment options with patients. PMID- 14741967 TI - Attitude of Arabian Gulf University medical students towards psychiatry. AB - INTRODUCTION: The attitude of medical students towards psychiatry has been studied extensively in the developed world. The inability to attract medical students to specialize in psychiatry has always been a serious challenge to psychiatric recruitment. OBJECTIVE: This study evaluates the attitude of medical students towards psychiatry and identifies associated factors. METHODS: Medical students from years 1, 4 and 7 at the College of Medicine and Medical Sciences (CMMS), Arabian Gulf University (AGU), constituted the study sample. One hundred and twenty two (87.1%) medical students completed the Attitude Towards Psychiatry 30 (ATP-30) scale and the study questionnaire. RESULTS: The attitude towards psychiatry was moderately positive (mean ATP score = 105.79, SD = 13.34). Twenty two (15.7%) students selected psychiatry as one of the top three career choices. Fifty five (72.3%) of the pre-clinical students agreed that teaching at the college is good or acceptable, compared to 11 students (44%) from the clinical phase. Female, younger and junior students had more positive attitudes than male, older and senior students. Factors such as exposure to material related to psychiatry, having a close relation with psychiatric illness and having a romantic relationship with a psychiatric patient were significantly related to the ATP score. CONCLUSION: The results of the study did not support the hypothesis that the greater the exposure to psychiatry, the higher the ATP. Possible explanations for the low attraction of graduates to psychiatry in spite of the initial high ATP and ways to overcome this problem are discussed. PMID- 14741968 TI - Students' and tutors' perceptions of problems in PBL tutorial groups at a Brazilian medical school. AB - INTRODUCTION: There are few published studies that address the problem of dysfunctional tutorial groups. Most studies are restricted to student or faculty opinions separately and to specific aspects affecting the tutorial group function. This study examined teacher and student perceptions of frequency and importance of problems observed in tutorial groups in a new PBL program. METHODS: Tutors (n=30) and students in the second (n=75) and third (n=53) year completed a questionnaire at the beginning of the 1999 academic year. The questionnaire had 33 items grouped as seven "factors" related to tutor performance, feedback, assessment, educational resources, student performance, educational problems and external factors RESULTS: The most important problems identified were related to tutors (mainly in training aspects) and students (mainly in problem discussion). Students and feedback (quality) were the most frequent. There were statistically significant differences between tutors' and students' (higher) and between second and third year (higher) students' perceptions of different factors. CONCLUSIONS: (1) Marilia Medical School (FAMEMA) has problems in the tutorial group function mainly related to contributions of students and tutors. (2) Students' and tutors' opinions, as well as those of second and third year students, differ and therefore all need to be consulted to solve tutorial group problems. (3) It is necessary to develop a better student training program and also to improve the tutors training program. (4) There is a need for continued evaluation of problem based learning at FAMEMA. We must look at perceptions of students from all years. PMID- 14741969 TI - Development of family medicine education in Moldova with Carelift International. AB - BACKGROUND: As in other former Soviet republics, Moldova's health system has been dependent upon multispecialty and hospital care. The government has undertaken a planning process to develop a primary care-based system utilizing family physicians. Carelift International and Moldova State Medical and Pharmaceutical University joined together to design an educational program to help create a family medicine specialty in the country. METHODS: Introductory concepts were incorporated into a workshop co-sponsored by the World Health Organization, Carelift International, UNICEF and Moldova Ministry of Health. Faculty teams participated in Carelift's 8-week US program, comprising a range of topics in family medicine: educational development at all levels, public health applications, health care organization, insurance, financing, and technology. Training also included 1 week in Finland, a fellowship in Lithuania, an in country workshop on rural health, and a supplemental 5-week US immersion program. OUTCOMES: A Department of Family Medicine was established, and a residency program instituted. It has already been strengthened with a 2-week introduction to the specialty, and rotations in family practice centers. Continued improvements and updates are planned. Urban and rural model family practice centers serve concurrent purposes of teaching, demonstrating and health care. Carelift shipped equipment for the principal center and a department library, and is equipping a teaching family practice center near the university. The Society of Family Physicians of Moldova was founded. The introduction of family medicine as a discipline into the health system of Moldova could be a valuable model for other former Soviet republics. PMID- 14741970 TI - Why health educators need epidemiology. AB - The aim of health education is to encourage health behaviors that promote a better quality of life and longer life expectancy. In the late 1960s, universities in the US began offering degree programs in health education. Most programs today require that at least one class be taken in epidemiology, where epidemiology involves the study of the distribution and determinants of disease frequency in human populations. In recent years, several competency areas have been set forth for health educators by the US National Commission for Health Education Credentialing. This paper specifically describes how training in epidemiology provides health educators with the ability to satisfy, in large part, these competency areas. The intent of this paper is to clarify to students and advisors of health education the rationale for requiring course work in epidemiology, as well as to emphasize that epidemiology is the cornerstone to all health education, whether conducted by physicians, nurses, or formally trained health educators. PMID- 14741971 TI - Community service, learning and the medical student. AB - CONTEXT: Medical students need learning experiences outside the classroom, clinic room and hospital room if they are to become integral parts of the communities in which they will practice medicine. Service-learning incorporated into the traditional medical school curriculum can provide a vehicle to accomplish this goal, and provide an avenue to enhance the professional development of the physician in training. METHODS: This paper describes efforts to incorporate community service and service-learning into a traditional medical school curriculum. The unique nature of our location in a rural state with several required rural rotations with our local community partners has facilitated this effort. CONCLUSION: Incorporation of service-learning into a medical curriculum can be accomplished and will enhance the professional development of the students. PMID- 14741973 TI - An interview of Hafiz el-Shazali Osman. Interview by Jane Westberg. PMID- 14741972 TI - Evaluating community-based health professions education programs. AB - This paper assumes the reader (1) has little knowledge about program evaluation, and (2) is interested in evaluation to improve a community-based health professions education program. There are other important and useful approaches that can be used to address an evaluation of a community-based health professions education program, and readers are encouraged to explore them--they appear in health education, public health education, in evaluation, and in program theory literature. The paper is organized around a group of questions as a reference or organizer for the reader. These include topics like why evaluation is wanted, what kinds of questions can be addressed through evaluation, who stakeholders are, who should conduct the evaluation, what methods can be used, and how to analyze data and report results from the evaluation. In the paper, I have attempted to include examples that are related to community-based health professions programs. Finally, the paper ends with the recognition that there is much more to learn in the field of evaluation and suggestions for ways to continue pursuit of knowledge in this topical area. PMID- 14741974 TI - An interview of Rosalina Batista. Interview by Jane Westberg. PMID- 14741976 TI - Institutional R2M: good or evil? PMID- 14741975 TI - An interview of SNO Chairman Stefan de Greef. Interview by Jan van Dalen. PMID- 14741982 TI - Developing appropriate community-based postgraduate training in a developing country. AB - The School of Medical Sciences, KNUST, established in 1975, indicated its educational orientation to a community-based and community-oriented system using the problem-based learning approach. Falling victim to the phenomenon of brain drain, the School developed a postgraduate medical curriculum which takes into account the needs and demands of national governments and communities for quality specialist care and equitable distribution of existing health facilities. There is an innovative community-based fourth year for obstetrics and gynaecology, ophthalmology, otorhinolaryngology, paediatrics, surgery and medicine. After attainment of the Part I Examination, residents are sent to Ulm, Maastricht, Britain and the United States for clinical attachments for a period of 3 to 6 months before returning to sit for their Final Part II for the Fellowship of the West African College of Surgeons or Physicians (FWACS, FWACP). By ensuring that the Final Examinations take place after the elective attachment, none of the new breed of specialists so far produced has been lost to the Western world. The strengths and weaknesses of the programme are discussed. PMID- 14741984 TI - A model for dental hygiene education concerning the relationship between periodontal health and systemic health. AB - PURPOSE: To develop a format for educating the appropriate health care professionals as to the relationships between periodontal inflammation and increased risks for poor diabetes control, cardiovascular disease, cerebrovascular disease, pre-term low birth weight, pneumonia and gastric ulcer reinfection. MATERIALS & METHODS: Dental hygiene students in the Advanced Periodontology curriculum were instructed to review current literature regarding the increased risk for systemic health problems when periodontal inflammation is present. Abstracts of the reviewed material were then presented in group setting to all course participants. For each systemic entity (diabetes, cardio/cerebrovascular disease, adverse pregnancy outcome, pneumonia, gastric ulcer) literature-based evidence of periodontal disease's association, affect, pathogenesis, validity and clinical significance was determined. Consensus statements for each entity were developed and used as a basis for clinical interpretation. Following this, patient health-history materials were developed to obtain the necessary information from patients while educating them about the increased risk for systemic health problems when periodontal inflammation is present. Lastly, correspondence materials were developed to alert managing physicians and medical auxiliaries about the increased risk for systemic problems in their patients who may present with periodontal inflammation. A methodology which medical personnel can use to quickly screen for the presence of periodontal inflammation in at-risk patients was also developed in these correspondences. CONCLUSION: An educational model and clinical materials were developed which are aimed at alerting patients, dental and medical personnel to the increased risk for systemic health problems when inflammatory periodontal disease is present. PMID- 14741983 TI - Centres for Clinical Education (CCE): developing the health care education of tomorrow--a preliminary report. AB - A new interdisciplinary concept of medical and health care education has been introduced at Karolinska Institute in collaboration with the County Council in Stockholm under the motto Learning together to be able to work together. Centres of Clinical Education are built up in four major hospitals to promote meeting places during clinical education for students from different categories. During a three-year project more than 5000 students from four educational programmes have been involved--medicine, nursing, occupational therapy and physiotherapy. The project started in 1998 and will turn into regular activities in 2002. The Centres consist of three parts. First is the Clinical Training Ward, a ward without patients where manual skills as well as skills in communication are taught, practised and videotaped. Second is the Clinical Education Ward, a student-driven ward where students during two-week periods experience their own professional roles in day-to-day work and learn how to work together. Third is the Multidisciplinary Team, where teachers from the four programmes plan and provide opportunities for students to learn together. Opportunities to meet and learn together have promoted a wider understanding of each professional in health care teams among students and staff. The Centres of Clinical Education provide excellent opportunities to bridge professional borders and to coordinate undergraduate studies and clinical reality. PMID- 14741985 TI - Audit as an educational package for family medicine residents in primary health care (PHC) rotation. AB - BACKGROUND: The Saudi Board of Family and Community Medicine requires residents of family medicine to perform an audit during PHC rotation. The aim of the project is to introduce the residents to their future responsibilities towards improving health services in PHC setting. THE PACKAGE: A series of small group discussion sessions introduces the concept of audit to residents. Each of them is then asked to audit one aspect of his work while in the rotation. The resident is assigned to a facilitator (assessor). At the end of the project the resident has to submit his work in a predetermined format. This format aims at helping the resident to comprehend his work, be concise and help the assessor to evaluate the work with considerable ease. RESULTS: The package is evaluated through regular opinion surveys that provoke participants' satisfaction with the experience. It has resulted in assessor and resident satisfaction and has facilitated some change in our teaching practice. This paper will highlight some examples to demonstrate residents' attainment of managerial skills. CONCLUSION: This package enhanced residents' awareness of audit and increased their managerial skills. Services in the health centers, which these residents had tackled, were improved as well. PMID- 14741986 TI - Changing educational paradigms to prepare allied health professionals for the 21st century. AB - CONTEXT: Inefficient and ineffective health care delivery has been of recent concern to most stakeholders in the process. Care provision systems will improve when care providers are educated to function as team members and to demonstrate competencies required for practice in diverse, demanding, and ever-changing environments. GOAL: In one School of Allied Health Sciences, faculty members from nine departments united to create an interdisciplinary curriculum designed to foster the achievement of common competencies essential for success in the workplace. APPROACH: Members of a Curriculum-2000 Task Force collaborated to: (1) review current literature, (2) articulate a set of common competencies across several disciplines, and (3) produce a proposal for achieving and measuring competencies in an interdisciplinary manner. CONCLUSION: Individuals from various disciplines can come to consensus about competencies that graduates should achieve. Such consensus is the first step in the direction of implementing a curriculum based on interdisciplinary competencies. PMID- 14741987 TI - Improving patient-centred medicine: a preliminary experience for teaching communication skills to Italian general practitioners. AB - INTRODUCTION: A key concept for general practice nowadays is that of patient centred medicine. In this model the physician's aim is to integrate the patient's experience of illness with the conventional understanding of disease, trying to reconcile the patient's agenda with his/her own. This paper describes a preliminary experience of a CME course on patient-centred medicine in Italy. AIM AND METHODS: The article focuses on a 7 hour course for teaching patient-centred medicine to Italian general practitioners. Assessment of the course was done both in terms of learner satisfaction and efficacy. Learner satisfaction was evaluated by a questionnaire with a 6-point Likert scale and course efficacy by a pre/post paper-and-pencil test. FINDINGS AND DISCUSSION: The pilot course on patient centred medicine seems to obtain high satisfaction in participants. Furthermore, an increase in competence with regards to patient-centeredness resulted after the course. The pilot study represents the first Italian CME seminar on patient centered medicine. Results obtained both in terms of satisfaction and efficacy suggest that the CME course is a valid educational tool. The opportunity to extend the experience to a higher number of participants is therefore recommended. PMID- 14741988 TI - Implementing a community education program on stroke for health care providers and consumers. AB - Stroke remains a leading killer and cause of disability in the United States. The incidence of stroke appears to be increasing while new advances in the management of stroke continue to emerge. These realities emphasize the need to communicate advances to health care providers and consumers. However, educating health care providers and consumers in rural states is often difficult due to dispersed geographic distribution and lack of resources. This project, utilizing teleconferencing technology, brought an educational program on stroke management for providers and consumers into rural communities. Community hospitals, with teleconferencing capability, were identified throughout Vermont. Community providers and consumers were invited to participate in educational sessions using a variety of marketing methodologies. A multidisciplinary team designed two curricula (one for providers and one for health care consumers) on the management of stroke. A total of 211 health care professionals and 122 consumers participated in the program. Education session evaluations suggested that the program either met or exceeded participants' expectations for the majority of programs. This project demonstrated that academic centers can provide quality continuing medical education for their rural communities using teleconferencing technology. Experience with this program suggests that there are three key elements for success: adequate planning time, communication on multiple levels, and strong marketing strategies. PMID- 14741989 TI - Clinical governance and the development of a new professionalism in medicine: educational implications. AB - This paper discusses the need for doctors to further develop the use of professional learning in the workplace as an effective method of coping with change. It uses the recent introduction into the UK, by the government, of clinical governance as an example of how professional learning can help doctors both cope with this imposed change and recognise their ongoing need to undertake a different and changing relationship with society, and it lays the foundations for coping with further change throughout their careers. PMID- 14741990 TI - Residents as teachers: outcomes of a brief training programme. AB - INTRODUCTION: Residents (RMOs) teach medical students in hospital wards, yet little evidence of efficacy is available for this activity. This study undertook to test the effectiveness of RMO teaching, and to delineate the principal barriers encountered. METHODS: Medical students in their fourth year were assessed at baseline for clinical examination skills in ophthalmoscopy, neurological examination of the legs, and rheumatological examination of the hands. One group of RMOs (n=6) taught ophthalmoscopy to the students attached to their unit, while the second group of RMOs (n=6) taught leg examination to their students. The third examination skill was not taught, but was used as a control intervention. The students were evaluated in all three skills by an observed, structured clinical examination at the beginning and end of their six-week attachments. RESULTS: There was a significant improvement in the skill of ophthalmoscopy in the intervention group (p<0.02), while the control group of students showed a decline in their abilities. The skill of examination of the legs improved after being taught, but not significantly. The RMOs universally reported that difficulty in finding time was a major barrier to their ability to teach medical students, and most set up additional teaching sessions rather than incorporate the teaching into their routine ward work. CONCLUSIONS: RMO teaching of medical students is effective, particularly for a skill which is novel. The difficulties arise in finding a suitable time for these important clinical teachers to transfer their skills. PMID- 14741991 TI - Effective poster design. PMID- 14741992 TI - An interview of Pedro Gordan. Interview by Jane Westberg. PMID- 14741995 TI - The Network: Community Partnerships for Health through Innovative Education, Service, and Research. Biannual meeting. Londrina, Brazil. October 2001. PMID- 14741998 TI - Reflections on change: educational and institutional implications of "regression toward the mean". PMID- 14741999 TI - Best practices in community-oriented health professions education: international exemplars. AB - INTRODUCTION: During 1998-2000, an international team of five researchers described nine innovative health professions education programmes as selected by The Network: Community Partnerships for Health through Innovative Education, Service, and Research. Each researcher visited one or two schools. Criteria for selection of these nine schools included commitment to multidisciplinary and community-based education, longitudinal community placements, formal linkages with government entities and a structured approach to community participation. The purpose of these descriptions was to identify key issues in designing and implementing community-based education. METHODOLOGY: Programmes in Chile, Cuba, Egypt, India, the Philippines, South Africa, Sudan, Sweden and the United States were visited. Before site visits were conducted, the researchers as a group agreed upon the elements to be described. Elements included overall institutional characteristics, curriculum, admissions practices, evaluation systems, research, service, community involvement, faculty development, postgraduate programmes and the school's relationship with government entities. Here I describe the common features of each of the nine programmes, their shared dilemmas and how each went about balancing the teaching of clinical competence and population perspectives. LESSONS LEARNED: Based upon an analysis of the cases, I present seven "lessons learned" as well as a discussion of programme development, institutionalization of reform and long-term implications for health professions education. The seven lessons are: (1) PBL and CBE are not seen as independent curricular reforms; (2) student activities are determined based upon sensitivity to locale; (3) health professionals need to work collaboratively; (4) there is a connection between personal health and population health issues; (5) population health interventions and treatment strategies need to be appropriate to local conditions; (6) graduates need to advocate for patients and the community in the public policy arena; and (7) organizational change takes a long time. CONCLUSIONS: Despite their differences, all nine exemplars are engaged in processes of organizational change. Schools are becoming more community-oriented and socially accountable, and all of these programmes have accepted two fundamental tenets: "take public money, give to the public" and "place matters". PMID- 14742000 TI - Curriculum change and strategies, past and present: why is it taking so long? AB - For the last 30 years or more the old war songs of medical education (community orientation, active learning, etc.) have been repeatedly ventilated in meeting after meeting, article after article. But of the present 1642 medical schools on the planet, only about 100 members of The Network and a few other have put these principles into practice, more or less. Obstacles and constraints are known. Is it the system, or the leadership, or the university culture, or the accreditation rules, or all of the above that are responsible for this lack of progress? Still, some overcame the obstacles, some removed the constraints. It would be useful to be better informed about HOW it was done. I suggest that Education for Health: Change in Learning & Practice (EfH) should present more articles explaining HOW successful schools managed to DO IT or WHY they were NOT able to DO IT. PMID- 14742001 TI - Assessing confidence and competence of senior medical students in an obstetrics and gynaecology clerkship using an OSCE. AB - CONTEXT: Assessment of clinical confidence and competence of senior medical students during an obstetrics and gynaecology clerkship using an objective structured clinical examination (OSCE). METHOD: A questionnaire was distributed to senior medical students (n=47)to assess pre- and post-OSCE self-perceived confidence and competence in five clinical skills: history taking, performing pelvic examination, interacting and communicating with patients, clinical reasoning and dealing with difficult patient situations. Pre- and post performance confidence levels were compared and correlated with OSCE scores. RESULTS: The five clinical skills were significantly interrelated (p=0.001). There was no significant difference in OSCE performances between male (n=16) and female (n=31) students. Pre- and post-OSCE confidence in performing pelvic examination was significantly higher in female than male students (p=0.01). Post OSCE confidence in clinical reasoning and dealing with difficult patient situations only were significantly increased in both groups (p=0.01 and p=0.02, respectively). Pre- and post-performance confidence levels were not significantly correlated to OSCE scores. CONCLUSION: Of five clinical skills rated, self confidence in clinical reasoning skills and dealing with challenging or complex patient problems only were significantly increased after an OSCE assessing competence. The content of some of our OSCE stations thus enhance confidence in these skills but psychometric and other characteristics of the OSCE such as duration and performance feedback mechanisms need further investigation. PMID- 14742002 TI - Teaching sociology within the speech and language therapy curriculum. AB - In the United Kingdom, the Royal College of Speech and Language Therapists suggests that sociology should be included within the speech and language therapy curriculum. However, in spite of this, sociology is seldom given priority. Although the role of sociology with the curricula of other professions has been discussed, the role of sociology within speech and language therapy has not. Given the contemporary climate of competence-based training, the position of subjects such as sociology is being compromised. This paper sets out three reasons in support of including sociology within speech and language therapy by drawing on the distinction between a "personal education" and a "semantic conjunction" model of the relationship between theory and professional practice. First, it is argued that sociology makes a valid contribution to an holistic approach to care--which is rapidly becoming the cornerstone of speech and language therapy practice. Second, this paper suggests that the inherent reflexivity within the discipline provides health professionals with an invaluable tool with which to engage in reflexive practice. Finally, it is argued that given the global emphasis on evidence-based practice, the study of sociology and sociological research methods equips therapists to interpret and conduct empirical research. This paper concludes by arguing that sociology should be viewed as an essential component of the speech and language therapy curriculum. PMID- 14742003 TI - Sociology (and all the behavioral sciences) have a place in all the health professions: commentary on "Teaching sociology within the speech and language therapy curriculum". PMID- 14742004 TI - Identifying medical school learning needs: a survey of Australian interns. AB - OBJECTIVES: To survey interns regarding their opinion of medical school learning needs for a range of core skills. METHODS: A random sample of interns practising in New South Wales, Australia, who graduated from the state's three medical schools were surveyed two-thirds of the way through their first hospital year. They were asked whether there was a need for further medical school education for each of 226 core skills. Skills were grouped into five themes: management of clinical conditions; clinical investigations; clinical procedures; core practice; and professional development. RESULTS: Frequency distributions weighted for age, gender and medical school background were calculated for each item. The 20 most frequently identified needs related to examinations of the eye and ear, nose and throat; managing uncooperative patients and difficult patient interactions; prescribing; writing not for resuscitation orders and death certificates. Also included were procedural needs related to ear, nose and throat; plastering and wound management; and needs for more education in the management of clinical conditions related predominantly to "acute" cases such as anaphylaxis and diabetic ketoacidosis. CONCLUSION: Interns were able to discriminate between their needs for different skills and identified many core skills for which they perceived there was a need for more medical school education. The implications for medical education are discussed. PMID- 14742005 TI - Attitudes of physicians and medical students toward nutrition's place in patient care and education at Ben-Gurion University. AB - OBJECTIVES: To describe attitudes of physicians and medical students at one medical school toward the role of dietetic treatment in patient care and toward adding nutritional education into the medical school curriculum. STUDY DESCRIPTION: A cross sectional survey was conducted at Soroka University Medical Center and Ben-Gurion University's School of Medicine. The attitudes of 67 physicians and 62 medical students toward nutritional treatment were determined using an attitudes questionnaire. RESULTS: Despite recommendations in medical treatment protocols to use dietary intervention as the primary treatment for several chronic diseases, physicians did not rate nutritional treatment as the most important treatment for these conditions. Students rated the importance of nutritional treatment significantly higher for each of the medical conditions presented than did practicing physicians (p=0.001). Almost 50% of the physicians reported not using nutritional treatment due to lack of time and awareness of the available options. Physicians and students rated the importance of nutrition education in the curriculum equally. Physicians who rated nutrition treatment as important also felt the need to add this subject to the medical education curriculum. CONCLUSION: Physicians and medical students agreed that dietary treatment and nutrition education are important. Our results suggest that there is good reason to introduce nutrition topics into medical school curricula. Improved nutritional knowledge in physicians would improve the teamwork capacity between physicians and dieticians in the realms of curative care and public health. PMID- 14742006 TI - The relationship between previous tertiary education and course performance in first year medical students at Newcastle University, Australia. AB - AIM: The purpose of this study was to identify the relationship between previous tertiary education background and the performance of first year medical students at Newcastle University, Australia. Specifically, we examined degree type (i.e. arts, science, allied health, nursing or other professional backgrounds), level of degree completion (fully or partially completed ), academic achievement ( grade point average) and whether or not students had postgraduate qualifications. The relationship between age and gender was also examined. METHOD: All students admitted to the medical course from 1990 to 1998 with previous tertiary education experience who entered via the "standard" entry pathway and sat the end of year examinations were eligible for the study (N=303). The outcome measures were the results of first assessment ("satisfactory" versus "not satisfactory") and final assessment of the first year ("satisfactory" versus "not satisfactory"). Logistic regression was used to examine the relationship between predictor variables and outcomes. RESULTS: In relation to first assessment results, students with a nursing and arts background were significantly more likely to receive a "not satisfactory" assessment (RR=3.9, 95% CI: 1.6- 7.7; RR=2.9, 95% CI: 1.2-6.8, respectively), as were females (RR=1.8, 95% CI: 1.1-3.5) and students with a grade point average of less than a distinction average (RR=2.8, 95% CI: 1.6-5.2). At final assessment, students with a nursing background and those with a less than distinction average were more likely to receive a "not satisfactory" result (RR=20.7, 95% CI: 3.5-123.9 and RR=4.0, 95% CI: 1.2-13.9, respectively); consequently, they were required to repeat first year. CONCLUSION: Our research suggests that there are some medical student groups who encounter more academic difficulties than others in first year. Identifying these students can assist medical schools to focus academic support appropriately. PMID- 14742007 TI - Integrating global health and medicine into the medical curriculum. PMID- 14742009 TI - An interview of Ron Richards. Interview by Jane Westberg. PMID- 14742008 TI - Meeting the challenge of a changing teaching environment: harmonize with the system or transform the teacher's perspective. AB - The beliefs that teachers hold about the appropriate roles and responsibilities of teachers shape the ways they teach and the ways they think about teaching. In this paper I describe four teaching roles based on a taxonomy that I've recently developed. Teachers who are guided primarily by the Content Expert Role view themselves as experts who serve as resources, like books or pictures. Teachers who are guided primarily by the Performance Role view themselves as agents who make learning happen by transmitting information or shaping students. Teachers guided primarily by the Interactive Role view themselves as guides who facilitate learning by interacting with learners. And teachers guided primarily by the Relational Role view themselves as engaged in relationships with learners for the purpose of helping them. Using examples taken from the health sciences I explain how each of the four teaching roles might succeed or fail depending upon the position that it occupies within a teaching-learning system. When teaching is viewed as part of a system, not as something a teacher does to a learner, teachers are successful if their particular contribution to the system is essential to the learning system. I also describe the process whereby teachers expand their belief system to include more roles. Such changes in belief systems are major shifts that qualify as "perspective transformations". Perspective transformations take place slowly and are typically attended by strong emotions. I end this paper with advice to teachers regarding ways they can harmonize with the educational system or face the challenge of perspective transformation. PMID- 14742011 TI - The search for HR continues: WHO's General Programme of Work 2002-2005 and the World Health Report 2001 on mental health. PMID- 14742013 TI - A heartfelt appeal: we need far more awe and wonder in our teaching! PMID- 14742014 TI - Toward a policy agenda for community-campus partnerships. PMID- 14742015 TI - Promoting collaborations that improve health. AB - There is growing interest and investment in health-related collaboration in the United States. In an environment characterized by increasingly complex health problems, substantial resource constraints, and a fragmented health system, public and private organizations as well as communities are recognizing that most objectives related to health and health care cannot be achieved by any single person or organization working alone. Partnerships that bring together diverse people and organizations have the potential for developing new and creative ways of dealing with today's turbulent environment. Despite its potential advantages, collaboration also presents daunting challenges. Further, documenting the effectiveness of partnerships in improving health and well-being has been difficult. Given the significant difficulties of collaboration and the lack of evidence of its effectiveness, questioning whether the investment in health partnerships is justified seems reasonable. In this paper we address this question by illustrating the connective power of collaboration. We describe how collaboration, by connecting individual-level services, broadening community involvement in population-based health strategies, and linking individual-level services and population-based strategies, can improve the health of communities. We then discuss activities that could assist partnerships in reaching their collaborative potential and conclude by presenting the most compelling reasons for pursuing collaboration. PMID- 14742016 TI - Service learning: integrating student learning and community service. AB - Health professions education is directly effected by changes in health care service delivery and financing systems. In the United States, as the health care industry increasingly shifts to a market economy, service delivery venues are moving away from acute care facilities and into community-based settings. Additionally, there is increased emphasis on primary prevention programs, often provided in public health settings. For health professions programs that traditionally provide clinical training in hospitals and long-term care facilities, there are unique challenges associated with identifying new venues in order to insure that students are exposed to a wide variety of patients with a range of chronic to acute disease conditions. One set of tools that has demonstrated usefulness during these kinds of transitions is service learning. This teaching methodology emphasizes increased partnership with clinical training sites, extensive orientation to patient populations and community resources, structured reflection and instilling the ethic of service in future health care providers. Although this article describes utilization of service learning in the context of current conditions in the United States, we hope that the principles presented here can be readily adapted in any setting. PMID- 14742017 TI - Community-based participatory research: policy recommendations for promoting a partnership approach in health research. AB - Community-based participatory research in public health focuses on social, structural, and physical environmental inequities through active involvement of community members, organizational representatives, and researchers in all aspects of the research process. Partners contribute their expertise to enhance understanding of a given phenomenon and integrate the knowledge gained with action to benefit the community involved. This article presents key principles of community-based participatory research (CBPR), discusses the rationale for its use, and provides a number of policy recommendations at the organizational, community and national levels aimed at advancing the application of CBPR. While the issues addressed here draw primarily upon experiences in the United States, the emphasis throughout this article on the establishment of policies to enhance equity that would serve both to increase the engagement of communities as partners in health research, and to reduce health disparities, has relevant applications in a global context. PMID- 14742018 TI - Student leadership and activism for social change in the US. AB - As the US health-care system continues on a path toward greater patient and provider dissatisfaction and decreasing access to primary health services, there is a growing need for leadership among tomorrow's health professionals. Students of today must acquire the skills to lead the United States toward solutions that will offer universal access and eliminate disparities. As examined in the US, there are two main modes for students to learn these meaningful skills; curriculum- and institution-based leadership development and organization-based leadership development. In this paper we review these two methods from an American perspective and discuss ways that educational institutions can support student leadership development. In addition, we address ways in which non governmental organizations can provide opportunities to foster student leadership. Lastly, we offer recommendations for US policy change at institutional, local, state, and national levels to help achieve the goals stated above. PMID- 14742019 TI - Working with our communities: moving from service to scholarship in the health professions. AB - CONTEXT: As faculty at health professionals schools have become increasingly engaged with their communities in partnerships to improve health, new questions have arisen about faculty rewards for such activities. To sustain the community work of their faculty, institutions need to reconceptualize faculty rewards, promotion, and tenure that are relevant to community activities. HISTORICAL PERSPECTIVE: Scholarship has evolved since the 17th century from a focus on character-building to the practical needs of the nation to an emphasis on research. In 1990, Boyer proposed four interrelated dimensions of scholarship: (1) discovery; (2) integration;(3) application; and (4) teaching. The challenge became the development of criteria and innovative and creative ways to assess community scholarship. CURRENT MODELS FOR COMMUNITY SCHOLARSHIP: This paper reviews four evidence-based models to document and evaluate scholarly activities that are applicable to community scholarship. PROPOSED MODEL FOR COMMUNITY SCHOLARSHIP: We propose a new model for community scholarship that focuses on both processes and outcomes, crosses the boundaries of teaching, research, and service, and reshapes and integrates them through community partnership. We hope this model will generate national discussion about community scholarship and provide thought-provoking information that will move the idea of community scholarship to its next stage of development. PMID- 14742020 TI - Public policies to promote community-based and interdisciplinary health professions education. AB - CONTEXT: Many rural and inner-city communities in the United States have persistent shortages of health professionals. In addition, health services are increasingly delivered in community-based settings by interdisciplinary teams. Yet, health professions students in the US continue to receive most of their training in urban hospitals. OBJECTIVE: To assess the extent to which national and state government programs in the US that fund health professions education provide financial resources for community-based and interdisciplinary education in the health professions. METHODS: Literature review. FINDINGS: Most national and state government funding provided to health professions schools and clinical training sites in the US is not targeted to community-based and interdisciplinary education. Nationally, the Bureau of Health Professions, however, does administer some targeted grant programs. In addition, a number of states are addressing these needs through targeted appropriations to health professions schools and Medicaid payments to clinical training sites. RECOMMENDATIONS: The US experience with government funding of health professions education suggests several questions that policymakers in other nations should consider and several principles for developing effective policies to promote community-based and interdisciplinary education. PMID- 14742021 TI - Town and gown in America: some historical and institutional issues of the engaged university. AB - The engaged university is a descriptive term for linking the different perspectives of university/community partnerships in higher education in the United States. While interest in this idea comes from recent events and processes, there is an historical background for university/community relations that offers important suggestions on the issue related to institutionalizing this concept. The history includes the original religious founding of US institutions, the establishment of the land-grant system of colleges, and the expansion of the mass education system in US universities. The development of community-based research disciplines, dating from the time of the settlement house movement in America, provides support for university/community collaboration. While some faculty and administrators support this idea, others are more critical and do not see it supporting the strategic interests of their institutions or disciplines. Comprehensively supporting the engaged university at an institutional level means changing the way the academy operates and including community concerns in research, teaching and administrative decisions. This article explores some of these issues and some possible solutions. PMID- 14742022 TI - Legislative advocacy for health professions educators. AB - Because much of health professions education in the US is publicly financed, the actions of politicians have profound effects on the organization of health professions education. The success of health professions education programs, therefore, depends in part on the ability of educators to advocate for change in the legislature. Successful legislative advocacy requires a general understanding of the legislative process and the needs of politicians combined with effective communications strategy. The tools of individual legislative advocacy include position papers, letter writing, politician meetings and visits, and using the media. Professional associations advocate on behalf of their members through coalitions, key contact programs, grassroots campaigns, and lobbyists. Successful legislative advocacy depends on credibility and the development of long-term relationships with members of the legislature. The process of legislative advocacy is straightforward and should be viewed as an integral part of health professions education. PMID- 14742023 TI - Evaluating a community-based multiprofessional course in community health. AB - PURPOSE: To evaluate student response to a community health course taught using a small-group, interdisciplinary, service-learning approach. METHOD: Student evaluations for the course were reviewed for a 3-year period (1994 1997). RESULTS: Student evaluations of the course improved over the 3-year period. A total of 60-76% of the students indicated that they preferred the small-group experiential approach to lectures. Examination of evaluation scores for individual small groups showed that some small groups gave the course very high ratings, while others found the experience inadequate. CONCLUSION: A course in community health is best taught in the community rather than the classroom. A small-group approach may result in a course with considerable variation among groups as a result of variations in community receptivity, faculty skills, and perhaps other factors. PMID- 14742024 TI - Educating health professionals in a community setting: what students value. AB - CONTEXT: Health care and health professions education has been shifting into community settings. Hawai'i participated in the Community Partnerships for Health Professions Education program, an initiative funded by the W.K. Kellogg Foundation that aimed to develop educational collaboration between community health centers, their respective communities and health professions schools. OBJECTIVE: In 1998, after completing 5 academic years in this program, former students from one community health center site were polled to begin exploring those components of their experience that were most valuable to their subsequent health care practice. METHOD: A survey was mailed to 65 former students, asking three questions: (1) what three components of your Ke Ola O Hawai'i experience (Wai'anae site) had the most impact on your practice;(2) is there anything else you would like us to know; and (3) where do you see yourself in 5 years? FINDINGS: Thirty responses were received for a 46% response rate. Students identified three components as having the greatest impact: (1) the multiprofessional approach to health care and learning, (2) the community setting/contact, and (3) understanding the culture of the community. A fourth component that was also revealed was the impact this experience had on their personal and professional growth. Most students reported employment or plans to practice in a community-based, rural, or underserved area. CONCLUSION: These responses introduce ideas for sustaining a community-based multi-professional curriculum that is relevant to current health care practice. PMID- 14742025 TI - Application of "VITALS": visual indicators of teaching and learning success in reporting student evaluations of clinical teachers. AB - CONTEXT: At the College of Medicine and Medical Sciences, Arabian Gulf University, Bahrain, a system has been introduced in which clerkship students evaluate clinical faculty using Visual Indicators of Teaching and Learning Success (VITALS). OBJECTIVE: To describe the use of VITALS in reporting student feedback on teaching and learning effectiveness of clinical faculty in the clerkship. DESIGN: Descriptive study. SUBJECTS: A total of 210 clerkship students evaluated 76 clinical tutors over a period of 3 years. Feedback was also obtained from seven programme managers and one supportive staff member. METHOD: Nine indicators of effective clinical teaching were identified through a literature search. Students individually reported on clinical faculty teaching capabilities using a 5-point, Likert-type scale. Cumulative reports of students' feedback on clinical faculty teaching were prepared using opposing bar graphs, reflecting perceived areas of strength or weakness in each teacher's performance. RESULTS: A total of 1450 evaluation forms were completed by 180 of 210 students (85.7%). VITALS graph representations of students' perceptions of clinical tutors were communicated to each clinical tutor at the end of each clerkship and academic year. Twenty-one students out of 53 who gave written comments were related to VITALS. They reflected a positive view of VITALS as a process or tool of faculty evaluation. Clinical faculty (18), programme managers(7) and supporting staff (1) gave comments indicating acceptance of the system. CONCLUSION: This preliminary study suggests that VITALS could be an effective tool for improving clinical teaching. It is acceptable to students, faculty and managers of educational programmes. The database reflecting their teaching and educational profiles were used to provide clinical faculty with constructive feedback. PMID- 14742026 TI - Can selection assessments predict students' achievements in the premedical year?: a study at Arabian Gulf University. AB - BACKGROUND: In a problem-based learning (PBL) program, students are encouraged to develop self-motivation, self-confidence, communication skills and problem solving skills. Measuring these attributes when selecting students into medical schools is a formidable task. Admission to medical school typically depends upon students' academic achievement in their prior education. In the past 3 years the College of Medicine and Medical Science (CMMS) at the Arabian Gulf University, which has a PBL curriculum, adopted an admission policy that utilizes final high school scores, a written admission examination in English and science, as well as a structured interview. OBJECTIVE: To determine the extent to which the admissions measures predict academic achievement in the first year of studies at CMMS. DESIGN: Prediction study of prospectively collected data. Final cumulative score for all subjects at the end of the first year was the main dependent variable analyzed. SUBJECTS: All students admitted to the college during the academic year 1998-1999. RESULTS: The written admission science examination scores had the highest correlation (r =0.663, p< 0.05) with the Year 1 final cumulative score. Although the admission interview focused on non-cognitive student attributes, which may or may not affect the students' academic performance, its score had a statistically significant, if low, correlation with the Year 1 final cumulative score (r =0.275, p< 0.012). Approximately 59% of the total variability of the Year 1 final cumulative scores could be explained by the admission examination scores in science and English and the high school scores. CONCLUSION: Procedures for selecting students who are most likely to succeed academically in the initial year at an innovative medical school deserve further study and probably should include both academic performance and non-academic attributes. PMID- 14742027 TI - Integrating a primer course in biostatistics into the haematology practicals of first-year medical students in India. AB - CONTEXT: In India, biostatistics in the medical curriculum is usually taught as a separate course, using either "imaginary" data or data from the literature. Knowledge of statistics is particularly important in the context of "evidence based medicine". OBJECTIVE: To evaluate the efficacy of a biostatistics course integrated into the practical haematology first-year medical course with the following special characteristics: (1) students learn statistics on data generated by and on themselves, (2) the course avoids mathematical computation,(3)the statistical exercises are linked to the learning objectives of the physiology curriculum, and (4) the course is without the threat of university examinations. METHODS: Statistical exercises were incorporated into specific haematology practicals with the aim of covering simple descriptive and inferential statistics. Statistics tests were administered, without prior information, to 60 first-year medical students before the biostatistics course, immediately following the course, and nine months later. Fifty-four students completed all three evaluations. RESULTS: Students had a poor knowledge of statistics at baseline. They substantially increased their scores in the statistics test immediately following the biostatistics course. These scores remained higher than baseline nine months after completion of the course, although there was a small decline in the absolute scores when compared with scores soon after the biostatistics course. IMPLICATIONS: Integrating biostatistics into other subjects in the medical curriculum may be an important addition to "stand alone" courses in biostatistics. PMID- 14742028 TI - Selective training and cross-year clinical tutoring as educational influences on generalist career choice. AB - CONTEXT: This study was undertaken with all 299 graduates of 12 consecutive classes (in 6 years) of a Brazilian medical school. PURPOSE: To appraise the relationships of gender, early preference for a career, the experience of selective training (a form of elective clerkship) and student preceptorship (cross-year clinical tutoring by peers), with the career choices of graduates. METHOD: Data were obtained at three points: at the beginning of medical studies, on career preference; at the end of medical studies, on selective traineeship and student preceptorship; and after graduation, on medical residency selected. Chi square tests and logistic regression analysis were done on the data of the graduates, grouped in terms of choice, or not, of frontline care specialties (internal medicine, pediatrics or obstetrics-gynecology). RESULTS: Findings are presented on 299 graduates, of whom 48.5% were female, and 53.8% preferred frontline care (FC) specialties at the beginning of the program. After graduation, 50.2% of the subjects chose FC specialties, among which two-thirds had kept their early preference for a specific specialty. Logistic regression analysis predicted 86.7% of the graduates' choices of FC specialties. Female gender, early preference, student preceptorship and, mostly, selective traineeship in the same broad area were significant factors. CONCLUSION: In this study a strong association was found between selective traineeship and career choice of FC specialties in relation to three additional predictive factors. It also revealed student preceptorship as a factor of potential educational significance in the career decision process and as a matter of institutional concern. PMID- 14742029 TI - Retraining and relicensing immigrant physicians: the Israeli approach. PMID- 14742030 TI - Behavioral problems in school-going children: implications for medical teachers in developing countries. AB - A large number of children suffer from behavioral problems during their development. Many of these problems are transient and may not even be noticed. At times, however, the extent of these problems and their overall effects on a child's development can be serious (Morita et al., 1993). Further, children may exhibit these behaviors in one setting and not in others (e.g. at home or in school, but not both). In developed countries, parents tend to seek advice for even minor problems, such as persistent thumb sucking, while in developing countries, major problems, even childhood schizophrenia, may go unattended. An awareness of the prevalence of these problems is important so that appropriate mental health services can be planned and provided for affected children, to improve their prospects for leading healthy, productive lives. Such awareness can help enhance the teaching of graduate doctors, equipping them to deal with these problems effectively. PMID- 14742031 TI - Helping learners become reflective practitioners. AB - CONTEXT: In too many schools in the health professions, students are given little or no opportunities to reflect systematically on their simulated or real experiences practicing the skills they need as clinicians (e.g. eliciting information and solving problems). In addition, they are given little or no help in learning the skills that are central to being reflective practitioners. RATIONALE: Reflecting alone or with the help of educators or others, students and residents can learn to identify and build on their existing knowledge, identify their biases and assumptions, integrate new understandings, and formulate generalizations that will enable them to make positive changes in what they do in future situations. Put another way, they can derive lessons from their experiences that will enhance their skills and enable them to provide better patient care. When learners reflect aloud on their insights and their strengths and deficiencies before their teachers give them advice or feedback, learners can make their own discoveries and have the dignity of identifying what they need to work on. The learners' self-disclosures provide educators with "diagnostic" information that can enable them to tailor their teaching to the learners' interests and needs. SUGGESTIONS: Educators can help learners become reflective practitioners by taking such steps as ensuring learners have worthy experiences on which to reflect; observing learners in action; scheduling times and places for reflection; building trust; determining the learners' prior experiences, comfort with, and attitudes toward reflection; ensuring learners understand the rationale and strategies for reflection; modeling reflection; clarifying the learning goals; encouraging learners to have questions to ask themselves; helping learners reflect on their thoughts, feelings, biases, and assumptions; helping learners consider other approaches; inviting learners to identify the lessons they've learned; and asking them to discuss how they intend to use these new understandings. PMID- 14742032 TI - An interview with professor Henk Schmidt. Interview by Jan van Dalen. PMID- 14742034 TI - How effective is problem-based learning? PMID- 14742036 TI - US Congress establishes new advisory committee on interdisciplinary, community based programs. PMID- 14742039 TI - Hearing what the patient is thinking: implications for care and education. PMID- 14742040 TI - The potential utility of problem-based learning in the education of clinical psychologists and others. AB - Clinical psychologists, like most health professionals, are in essence clinical problem-solvers. However, dealing with mental health problems may necessitate a greater relative reliance upon inductive clinical reasoning during the problem solving process. To develop a provisional problem formulation mental health professionals may have to make sense of the co-occurrence of complex and poorly delineated problems. Claims have been made, predominantly in the literature on medical education, regarding the utility of problem-based learning (PBL) for achieving aims central to the effective performance of this role. In this article, after characterizing clinical psychology and PBL, we briefly explore the benefits claimed for PBL and assert that the putative cognitive and interpersonal consequences of the approach may be particularly pertinent to mental health practice. Particular emphasis is placed upon the necessity of facilitating effective clinical reasoning, that is, teaching future practitioners how to, rather than what to, think about complex psychopathology. PBL is also considered in the wider context of models of experiential learning and methods for teaching problem-solving. Finally, future research questions are suggested which may provide answers relevant to the facilitation of effective clinical reasoning in all health professions. PMID- 14742041 TI - Evaluation of electronic discussion groups as a teaching/learning strategy in an evidence-based medicine course: a pilot study. AB - BACKGROUND: As course directors, we wished to incorporate small group learning into our Evidence-based Medicine course for students to get feedback on the development of a well constructed, researchable clinical question. Scheduling of these groups was problematic. We sought to evaluate computer-mediated communication as an alternative to face-to-face small groups. METHODS: Students were randomly assigned to either face-to-face small groups or asynchronous, electronic, small groups. Final examination scores were analyzed with an analysis of variance to determine if there were differences in student performance based on group type. Student survey items were analyzed using Fisher's Exact test to determine if there were differences in student attitudes based on group type. RESULTS: There were no significant differences found in overall student performance. Significant differences in student attitudes were found to exist with respect to: (1) participation in discussions, with face-to-face groups reporting greater participation; (2) putting more thought into comments, with electronic groups reporting more thought put into comments; and (3) difficulty relating to other students in the class, with electronic groups reporting more difficulty. DISCUSSION: We found electronic discussion groups (computer-mediated communication) to be a viable teaching/learning strategy with no adverse effects on student performance or attitudes. PMID- 14742042 TI - Commentary: Pilot studies. AB - As its label implies, a pilot study or project is a trial of an idea that may be a new instructional model, innovation, technique, or curriculum reform. Originally it was used for trials of large-scale research designs. Pilot studies and their close relatives, feasibility studies and demonstration projects, have had a varied pattern of use since the 1960s. Kamin et al. (2001) have provided us with an excellent example of how a pilot project can be undertaken to identify the strengths and weaknesses of an instructional model prior to widespread and/or permanent adoption of its use. PMID- 14742043 TI - One year's experience with a program to facilitate personal and professional development in medical students using reflection groups. AB - PURPOSE: (1) to integrate sociobehavioral science concepts into the early curriculum through a continuity ambulatory clinical experience in primary care, and (2) to expose students to a learning environment in which self-awareness and emotional development are nurtured in the context of dealing with the stresses of an early clinical experience. METHODS: Second-year students spent half a day twice monthly in a primary care community practice, kept a journal of their experiences, and attended biweekly 60-minute Reflection Groups designed to foster personal awareness and empathic witnessing. Analysis of journal entries and Reflection Group field notes identified stressors occurring during the students' clinical encounters. RESULTS: Three sources of stress are illustrated: the role and responsibility of the physician, death and dying, and racial issues. Reflection Groups provided students with opportunities to identify and describe stressors, to feel less isolated, to begin the process of self-awareness development, and to integrate behavioral and social science concepts into clinical practice. Our program incorporates students' early clinical experience with facilitated opportunities to reflect on the emotional challenges of becoming a physician. PMID- 14742044 TI - Songs and storytelling: bringing health messages to life in Uganda. AB - In villages without doctors and hospitals in the remote eastern Ugandan district of Pallisa, traditional birth attendants and mothers are solving the most serious health problems through the teaching power of songs and stories. The village's rich oral tradition was enlisted as the principal means not only for transmitting these important health messages, but also for supporting their practice throughout the community. Utilizing existing community traditions such as songs and storytelling offers culturally appropriate ways of enhancing the communications component of the health care system to make it serve the poor majority in a readily comprehensible, credible, affordable, and accessible form. These non-formal active-learning methods are highly compatible with and promotive of the general principles of primary health care, especially for their empowering, participatory and sustainable qualities. It is only a natural extension for health educators to more fully employ the use of the time-honored oral traditions of songs and storytelling as a vehicle for communicating health messages. For students in the health professions, awareness of these proven principles for engaging people at the local levels will contribute to more effective training, strategic program design, and advocacy. PMID- 14742045 TI - The anxieties of male and female medical students on commencing clinical studies: the role of gender. AB - CONTEXT: Many medical students experience considerable anxiety when starting hospital experiences. OBJECTIVES: To investigate the role of gender in this transitional experience. METHOD: A questionnaire study was conducted in 1992 and 1995 to compare female and male anxieties about clinical situations they anticipated encountering. The 31-item questionnaire listed potential anxiety provoking situations and requested the respondents' ratings of their reactions (from 1=not anxious to 4=very anxious). RESULTS: Differences in rankings between males and females were consistent between 1992 and 1995. More detailed analysis of 1995 data showed females had significantly higher totals; for 16/31 situations the difference was statistically significantly higher. For 4/31 situations male score was statistically significantly higher. Males ranked clinical situations involving intimate contact with patients significantly higher. Females scored situations involving interactions with consultants significantly higher than other situations and higher than did males. Females compared to males ranked 6/31 situations over five places different. CONCLUSIONS: Customisation of clinical introductory courses should be introduced. Earlier community-based clinical experience may help reduce non-productive anxiety. PMID- 14742046 TI - Learning by doing: teaching qualitative methods to health care personnel. AB - PURPOSE: We describe and assess the teaching of qualitative methods to postgraduate students using a case study from the Centre of International Child Health, Institute of Child Health, London, which trains mainly health personnel with developing country experience. As the majority of these students are practitioners rather than academics, the teaching focuses on combining theory with practice. We then analyse the results of the assessment of students about this approach and examine lessons learned from this experience. APPROACH: We present the format of a two-week course and the evaluation of this course by the students of the past four years. We describe the process of conducting a learning by-doing course, giving the day-to-day details of how the course is conducted. We also give examples of how this teaching is done. RESULTS: One indicator of the value of this course is its increasing popularity over the past three years. In 1997-1998, 11 students out of 20 took the course. In 1998-1999, 16 students out of 21 opted for this qualitative module. In 1999-2000, 12 students out of 17 chose this module. DISCUSSION: The lessons learned from this experience include challenges in how to present the teaching within the available time and having realistic expectations concerning course outcomes. We argue that a learning-by doing approach accomplishes the objectives of having students recognize the value of these methods for health systems research and giving them skills needed to use these methods. PMID- 14742047 TI - How do Australian doctors with different pre-medical school backgrounds perform as interns? AB - AIM: To assess whether there is any advantage to be gained with respect to performance in the first year of postgraduate medical training (internship) by selecting medical school candidates with different educational backgrounds. Specifically, we were interested in comparing the performance ratings of interns who entered medical school with secondary (directly from high school) or tertiary (at least one year of a university degree) level educational backgrounds. FOCUS: We compared the performance ratings of interns according to the subjects or degree undertaken at a secondary or tertiary level, respectively. The effects of age and gender were also examined to determine their influence on performance ratings. METHOD: All graduates (N=235) from the University of Newcastle Medical School, Australia who commenced their intern year in the state of New South Wales from 1993 to 1996 inclusive were eligible for the study. The outcome measure was a score derived from a valid and reliable clinical supervisor rating scale. Independent variables were level of previous educational experience (secondary or tertiary entry), and subjects studied by secondary level entrants (predominantly science or equal proportions of humanities and science) and degree undertaken by tertiary level entrants (arts or science or allied health or nursing). RESULTS: The records of 173 (73% of eligible sample) were included in the analyses. There were no significant differences between the mean ratings of interns with respect to previous educational background, subjects studied at secondary school or degree undertaken. Age and gender did not significantly affect performance ratings. PMID- 14742048 TI - A screening process predicts success rates in residency re-training among new immigrant physicians in Israel. AB - OBJECTIVES: To evaluate predictors of success in training programs for Israeli international medical graduates (IMGs) from the former USSR. SUBJECTS: IMGs up to the age of 45 who passed a national licensing examination and a Hebrew knowledge exam were eligible for participation. INTERVENTION: Each IMG participated in one of three programs that offered similar academic activities. In two programs there was a preliminary selection process and one program had supplementary teaching hours. MAIN OUTCOME MEASURES: Resident's self-evaluation of skills compared with evaluation by department heads. The evaluation included eight clinical skills, a general evaluation, and the passing rate for Phase I of the National Certification Exam. FINDINGS: A total of 176 IMGs, all graduates from medical schools in USSR who immigrated to Israel from 1990 to 1996, and 20 heads of the participating departments completed the questionnaires. The response rate was 64% among the residents and 71% among the department heads. All residents had a similar level of self-esteem regarding clinical skills and considered themselves equal to or better than Israeli-trained residents. The department heads did not concur with this assessment, particularly for residents who participated in the program with no preliminary selection. Participants in the programs with preliminary selection received better evaluations from department heads than those in the program without and had significantly greater success in the written part of the board certification examination (p < 0.05). DISCUSSION AND CONCLUSIONS: Re-training programs for IMGs should include a preliminary process for participant selection. The training program should be implemented in academic departments, in which case supplementary teaching hours are not required. PMID- 14742049 TI - Getting qualitative research published. AB - Translating research findings in health education into a publishable manuscript is challenging regardless of whether qualitative or quantitative methods are used. In this paper, we offer practical advice about how to successfully prepare and guide manuscripts based on qualitative research methods, in particular through the peer-reviewed journal publication process. Researchers trying to publish qualitative findings may face some unique challenges, given the field's current knowledge of qualitative methods, evaluation criteria, and conventional manuscript styles and length. PMID- 14742050 TI - An interview of Arthur Kaufman. Interview by Jane Westberg. PMID- 14742051 TI - An interview of Lisa Wallin. Interview by Jane Westberg. PMID- 14742056 TI - Influencing health behavior: why it matters; learning what to do. PMID- 14742057 TI - Educating doctors to provide counseling and preventive care: turning 20th century professional values head over heels. AB - Internationally, 20th century medical education concentrated on equipping new graduates with technical skills and pushing the frontiers of technological sciences to extend and enhance life in ways unimaginable in previous decades. In the 21st century, health services are expected to be characterized not by the "fix-up-when-things-go-wrong" type of care that 20th century physicians have become so good at, but by preventive care that can obviate much of the need for these fix-up services. Enabling doctors to deal with the different health care needs of future patients will require a values shift in medical education. The United States leads the world in per capita health care expenditure yet trails in many important measures of health status. It epitomizes many elements of both the good and the bad in current medical education that may be less obvious in other countries that are less wealthy, less technologically oriented, and less committed to individual freedoms. In this paper we use the US as a case study to argue the need for a fundamental shift in values away from the 20th century emphasis on disease, specialization and treatment, and towards health, generalization and prevention. We draw on data from the National Ambulatory Medical Care Survey to compare roles of primary care physicians and other office based medical specialties in delivering preventive health care. We also estimate the cost of providing preventive care in terms of physician time. Finally, we contemplate how medical education values must change in the US and other countries if 21st century physicians are to be prepared to meet the health care needs of their communities. PMID- 14742058 TI - Social determinants of (un)healthy behaviors. AB - Medical education has historically relied on the rational choice model as a vehicle for promoting health behavior change, and has largely overlooked the powerful relationships between social class and health behaviors. The rational choice model, which assumes that people can choose to pursue behaviors that are needed for their health, has some clinical utility, especially in some circumstances, but it runs the risk of missing key sources of influence and of blaming the victim. The biopsychosocial model provides an alternative basis for teaching about health behavior change. Health behavior needs to be understood in a broad social context, in which social class is recognized as playing a large part in shaping many people's health behaviors through multiple pathways, including limited opportunities for self-fulfillment, financial constraints, health beliefs, self-efficacy, stress, and social support. In addition to highlighting the limitations of the rational choice model, we illustrate how to integrate the socio-cultural context into teaching about behavior change. Specific curricular suggestions include exercises for: (1) increasing students' awareness of their own biases regarding unhealthy behaviors and individual responsibility for change; (2) enhancing knowledge of social factors that impact health; (3) building advocacy skills; (4) learning from patients; and (5) practicing counseling skills through role-plays. PMID- 14742059 TI - Spreading the word: Teaching health promotion to students from disciplines other than health. AB - Intersectoral collaboration has gained acceptance as a strategic approach in promoting health, based on the assumption that the main determinants of health are social, physical and politico-economic factors and not medical care utilization. However, the difficulties of collaborating intersectorally for better health have become apparent over the last two decades. This paper describes an attempt to address these difficulties through an awareness-raising educational initiative devised for undergraduate university students from disciplines other than health. The course aims to raise students' appreciation of the ways in which their future occupations could have an impact on the health of others through intersectoral collaboration and the creation of environments that are supportive of health. The evaluation of the course, which comprised a peer review process, a questionnaire seeking student feedback, and a task exploring students' ideas on how they could influence the health of others, demonstrated that students recognized the value of working intersectorally for better health. If intersectoral collaboration is really expected to be the way forward that many public health theorists claim, then this small but effective contribution should best be seen as one of a series of incremental building blocks leading to the desired effects. PMID- 14742060 TI - Teaching behavior change skills to first-year medical students: a small group experiential approach. AB - PURPOSE: Develop and evaluate a course to help first-year medical students learn about health-related behavior change by focusing on their personal health goals. COURSE METHODS: Students each identified two health-related behavior change goals for themselves. Lectures presented relevant content concerning behavior change. In small group, experiential sessions, students formed five-person teams and rotated positions as "patient," "doctor," "manager" and observers. "Doctors" and "patients" had one or two follow-up visits. Students evaluated their goal attainment and the value of their experience as "patient" and as "doctor." SAMPLE: 100 first-year medical students. RESULTS: Students chose exercise, nutrition, academic, interpersonal and psychological goals. Rating of the educational values and goal attainments were variable. Mean rating for educational value was 40%, and for goal attainment 55%. CONCLUSIONS: Experiential learning is valuable in introducing medical students to behavior change. Students learn from both the role of change agent and the role of "patient." Although the design of the course was cost-effective, with appropriate modifications considerably more impact could be gained from such a course. To improve this type of experiential learning we recommend careful attention to goal-setting, and more attention to developing the learners' feedback-giving and facilitation skills. PMID- 14742061 TI - A model for educating humanistic physicians in the 21st century: the new medicine, patient, and society course at Tel Aviv University. AB - BACKGROUND: The impact of the social and behavioral sciences on medical education has often been limited due to a variety of organizational, curricular and professional barriers. The new "Medicine, Patient, and Society (MPS)" program in Tel Aviv attempts to rectify this educational shortcoming by exploring new ways to help students acquire the knowledge, attitudes and skills needed for becoming humanistic physicians and for helping patients (and themselves) adopt healthy behaviors. To work toward this goal, this program integrates the biomedical and psychosocial aspects of health care, providing developmentally appropriate learning experiences according to levels of training, together with a variety of educational methods, including learner-centered approaches. OBJECTIVES: To implement and evaluate the MPS pilot program. METHODS: The MPS program uses a "seamless" model of behavioral science education. This integrated curriculum interweaves several elements: behavioral science topics (presented through multiple approaches), clinical experiences, practical medical skills, and an independent project. During the program's first year there is a strong focus on "health" rather than "disease," with activities designed to encourage healthy behaviors, including smoking cessation, stress management, birth control, AIDS education, life cycle and preventive health services. Assessment of the pilot for first-year students included standardized questionnaires, student focus groups, participant observation of educational activities, and committee feedback. RESULTS: Students' quantitative evaluations indicated high levels of satisfaction with the MPS program, but their qualitative evaluations revealed some concerns. Participant observations and focus groups added unexpected insights. Student concerns included performance fears, difficulties with "learner-centered" education, and incompatibilities between more traditional first-year courses and the MPS program. Long-term follow-up will be needed to determine the impact of this emphasis on health during the first year. We assume it serves as a helpful foundation for students before they focus on disease and its sequelae in their later years. PMID- 14742062 TI - Teaching children about health: an example of secondary gain in an academic community partnership. AB - CONTEXT: The Partners in Health Education (PHE) program is an elective that pairs first and second year medical students with local classroom teachers to promote health messages to students in kindergarten through grade eight. Designed with the primary goal of helping medical students improve their communication skills through the process of teaching children about health, the PHE program has secondary goals of supporting community teachers in their efforts to promote health and of teaching children about health and the prevention of disease and injury. This report contains the results of the assessment of program impact on the school children. METHODS: A total of 327 elementary grade students in 14 experimental classrooms and 13 comparison classrooms comprised the participants for the study. Students were individually interviewed twice over an eight-week period using a structured interview form designed to capture self-report information about health and healthy living. Repeated measures analysis of variance was conducted. The effect of interest in each case was the treatment x time interaction. RESULTS: There were significant treatment x time interactions for several measures of children's reported knowledge and attitudes about health. DISCUSSION: Although designed primarily to help medical students improve their communication skills, the PHE program produced a secondary gain such that elementary students in participating classrooms reported learning more about health than did students in comparison classrooms. Programs such as PHE can provide ways to meet the goal of helping children become empowered to take charge of their own health and to make healthy choices. PMID- 14742063 TI - Adoption of safe water behaviors in Zambia: comparing educational and motivational approaches. AB - CONTEXT: In the developing world, drinking water is an important route for transmission of diarrheal disease, a leading cause of morbidity and mortality in children. OBJECTIVES: In Field Trial 1 (FT1) and Field Trial 2 (FT2), the effectiveness of the behavior change approach known as motivational interviewing (MI) was compared to the standard practice of health education alone in initiating and sustaining safe water treatment and storage behavior among community residents. In Field Trial 3 (FT3), MI was compared with social marketing. DESIGN: Community surveys were conducted prior to local health promoter training and at follow-up. SETTING AND PARTICIPANTS: Low socioeconomic status peri-urban communities in Zambia were project sites. Local volunteer health promoters from communities were trained in an adaptation of MI for safe water treatment and storage. INTERVENTIONS: All health promoters received instruction in the causes and prevention of diarrhea. Health promoters in the experimental (MI) groups received MI training. MAIN OUTCOME MEASURES: FT1 and FT3 measured detectable disinfectant levels in stored household water. FT2 measured disinfectant sales. FINDINGS: No significant differences between the treatment groups were found in FT1. Subsequent MI training incorporated lessons learned from the previous trial and resulted in much higher purchase rates of the disinfectant (FT2) and levels of disinfectant in stored household water (FT3) in the MI group. CONCLUSION: MI appears promising for public health initiatives in the developing world. Further research to improve and refine the method is needed. PMID- 14742064 TI - Brief negotiation program for promoting behavior change: the Kaiser Permanente approach to continuing professional development. AB - BACKGROUND: Behavior change counseling is one of the most difficult and constant challenges faced by health providers. It has a significant impact on clinical outcomes as well as patient and provider satisfaction. By encouraging patients to participate in a partnership with health care professionals, Brief Negotiation offers techniques to motivate behavior change successfully. We review the key components of Brief Negotiation and describe how one large group model health maintenance organization was able to identify key staff members, develop educational opportunities and implement Brief Negotiation system-wide into standard care practices. OBJECTIVES: To expose a maximum number of health care professionals to a recommended model of behavior change counseling; to increase the satisfaction and confidence of health care professionals in counseling for behavior change; and to increase the likelihood of improved patient health outcomes. METHOD: Two departments created one-day, two-day, six-hour and one-to two-hour skill-based programs targeted to physicians, nurse practitioners, care managers, clinical health educators, behavioral medicine specialists, physical therapists, pharmacists and medical assistants. Practice protocols, strategic departmental alliances and intranet sites complemented the educational interventions. RESULTS: Over 1000 health care professionals have been exposed to the Brief Negotiation model in over two years. A mailed survey to graduates of the one- and two-day programs indicated that 67% of physicians and 79% of other health professionals felt more confident about working with patients on behavior change after having attended the Brief Negotiation program. CONCLUSIONS: System wide professional development requires multiple exposures to the Brief Negotiation model, considerable resources for curriculum development, training time and follow-up, and credible clinical trainers. Questions remain about the amount of training needed for long-term clinician behavior change and for improved health outcomes in patients. PMID- 14742065 TI - Improving physician-delivered counseling in a primary care setting: lessons from a failed attempt. AB - BACKGROUND: The high prevalence of behavioral risk factors for cardiovascular diseases demands innovative approaches to achieving behavior change. Primary care physicians are in an ideal position for offering such interventions. PURPOSE: To evaluate whether training of primary care physicians in counseling skills based on the Transtheoretical Model (TTM) leads to motivational and behavioral changes in their patients. METHOD: Seventy-four primary care physicians in Germany were randomly assigned to either an intervention condition (one day of training in TTM based counseling plus brochures matched to their patients' "stages of change") or a control condition (usual care). Baseline and 12-month follow-up data were collected from 305 of their patients who signed up for a health check-up. OUTCOME MEASURE: Patients' movements across the stages of change for smoking, diet, exercise and stress management. RESULTS: After 12 months, patients of physicians in the intervention group did not show more movement through the stages of change for any of the behaviors than did patients of control physicians. Additionally, there were no differences between groups in counseling frequency, counseling intensity, or patient satisfaction with counseling. CONCLUSIONS: A high dropout rate at follow-up and resulting "power" problems limit the possible conclusions. The high numbers of patients in early stages of change and the minimal improvement over time underline the need for improving motivational counseling skills of primary care physicians in Germany. In our study the dissemination of these strategies failed. We offer lessons we feel can be learned from this outcome. Further studies should focus on ways to enhance the process of educating physicians for implementing counseling strategies in primary care settings. PMID- 14742066 TI - A "Healthy Schools" program in Hong Kong: enhancing positive health behavior for school children and teachers. PMID- 14742067 TI - Is an arcade-style computer game an effective medium for providing drug education to schoolchildren? PMID- 14742068 TI - An interview of Abraham Joseph. Interview by Jane Westberg. PMID- 14742070 TI - Meaningful medical education is therapeutic for those afflicted with chronic disease. PMID- 14742074 TI - Communication skills are vital in all we do as educators and clinicians. PMID- 14742075 TI - Commentary: Research evidence will not be enough. PMID- 14742076 TI - Medical education, research, and scientific thinking in the 21st century (part two of three). AB - BACKGROUND: During the 20th century medical education has been largely preoccupied with discussions of the venues and methods for teaching. Little attention has been paid to what should be learned about the scientific paradigm underlying research and practice. A 17th century model has gradually produced an increasingly narrow, monocausal, reductionistic view of health and disease. Much good has resulted, but this "belief system" fails to accommodate or explain the meaning and impact on patients' health of diverse internal and external experiences and influences. During this period quantum mechanics and its ever expanding capacity to accommodate new information and enhance understanding have superseded Newtonian physics in much scientific thinking. METHODS: A broad range of historical and contemporary scientific literature is examined in support of four central questions in this three-part series: (1) Are there reasons to examine these matters now? (2) How is medical scientific thinking influenced by the general reorientation of science during the 20th century? (3) Are there reasons now to examine the impact of these changes on medicine? (4) Will a change of paradigm affect medical practice, research, and education? RESULTS: The extraordinarily productive contemporary biomedical model should be expanded beyond the physical and biological to incorporate meaningful information about how each patient's experiences impinge on health status. CONCLUSIONS: Family and other primary care physicians together with collaborators in the biological and behavioral sciences and other health professions should undertake rigorous research to establish the validity of the expanded paradigm espoused. Its impact on practice, research, education, and policies could be profound. PMID- 14742077 TI - Medical education, research, and scientific thinking in the 21st century (part three of three). AB - BACKGROUND: During the 20th century medical education has been largely preoccupied with discussions of the venues and methods for teaching. Little attention has been paid to what should be learned about the scientific paradigm underlying research and practice. A 17th century model has gradually produced an increasingly narrow, monocausal, reductionistic view of health and disease. Much good has resulted, but this "belief system" fails to accommodate or explain the meaning and impact on patients' health of diverse internal and external experiences and influences. During this period quantum mechanics and its ever expanding capacity to accommodate new information and enhance understanding have superseded Newtonian physics in much scientific thinking. METHODS: A broad range of historical and contemporary scientific literature is examined in support of four central questions in this three-part series: (1) Are there reasons to examine these matters now? (2) How is medical scientific thinking influenced by the general reorientation of science during the 20th century? (3) Are there reasons now to examine the impact of these changes on medicine? (4) Will a change of paradigm affect medical practice, research, and education? RESULTS: The extraordinarily productive contemporary biomedical model should be expanded beyond the physical and biological to incorporate meaningful information about how each patients experiences impinge on health status. CONCLUSIONS: Family and other primary care physicians together with collaborators in the biological and behavioral sciences and other health professions should undertake rigorous research to establish the validity of the expanded paradigm espoused. Its impact on practice, research, education, and policies could be profound. PMID- 14742078 TI - Choosing a career in primary care: the road not taken. AB - CONTEXT: Despite a mission statement and curriculum that are unique in our country in proposing to direct physicians to primary care (PC), the proportion of doctors graduating from Ben Gurion University (BGU) who choose PC is similar to that of other Israeli medical schools. OBJECTIVES, METHODS AND STUDY POPULATION: To investigate factors underlying our graduates' career choices we sent a questionnaire to six consecutive classes that had graduated from this medical school. We hypothesized that medical school was not the decisive factor influencing career choice. RESULTS: Returns were received from 135 graduates (54%). The nature of a specialty was the most important factor in choosing a career and in rejecting PC. Differences between primary care physicians (PCPs) and non-primary care physicians (NPCPs) were identified. PCPs emphasized factors relating to their personal lives. NPCPs emphasized the nature of a specialty in career choice. The most important factor in choosing PC was the physician-patient relationship and human aspects of medicine. Medical school was viewed as playing a minor role in career choice. PROPOSED INTERVENTIONS: Graduates proposed methods to increase the proportion of doctors choosing PC. These included: economic incentives; changing work conditions; strengthening contact with tertiary care; continuing medical education; and changing PC clerkships in medical school. CONCLUSION: The inherent nature of a specialty is central to career choice. In PC, the patient-physician relationship is central to physicians' career choice. PMID- 14742079 TI - Commentary: Research on specialty choice: the challenge is in the details. PMID- 14742080 TI - A computer-based epidemiological skills module for medical undergraduates in Nepal. AB - OBJECTIVE: To ascertain whether undergraduate medical students could acquire the basic competence needed to design, analyze and use a computer-compatible database in an epidemiologically meaningful way. SETTING: Since 1998 the Manipal College of Medical Sciences (MCOMS) of Kathmandu University (KU), Nepal has included in its undergraduate medical curriculum a training module that incorporates knowledge and skills in doing computer-assisted data analyses of epidemiological questions. The present study was designed around the implementation of this module. SUBJECTS: All 96 students of the seventh semester (first six months of the fourth year) of the MBBS course at the MCOMS. METHODS: The teaching-learning activities for the module were carried out mainly in six 2-hour sessions, for groups of 16 students at a time. The software used was EPI-INFO. MAIN OUTCOME MEASURES: Knowledge and skills of computer-assisted data analyses were assessed. In addition, feedback was obtained from the students and scored on seven dimensions. FINDINGS: The pre-test/post-test questionnaire score difference, which evaluated the knowledge component, was highly significant (t = 51.3, p < 0.001). In the skills assessment session, 83% were successful. The students gave high average scores on the satisfaction, small group learning environment, curricular relevance and evaluation dimensions; but the scores were relatively low on the time, other resources and confidence dimensions. CONCLUSION: The module was successful in achieving its objectives and can be implemented even under tight resource constraints. Our plans for improving upon this first run of the module are outlined in the paper. PMID- 14742081 TI - The Del Rio project: a case for community-campus partnership. AB - CONTEXT: Interdisciplinary teams of graduate health professions students and faculty were provided with experiential learning opportunities while assisting a small rural community address critical health-related issues. PROJECT OBJECTIVES: To establish an effective partnership with community leaders and area residents to assist in determining the feasibility of a new primary care clinic and to remediate a water borne disease threat. To create interdisciplinary clinical learning experiences and to develop future longitudinal learning opportunities, emphasizing primary prevention. To create a community-campus partnership with control originating in and sustained by the community. PARTNERSHIP DEVELOPMENT: An interdisciplinary team of health professions students and faculty worked with community leaders and residents to develop leadership skills, enhance infrastructure and coordinate efforts to address health concerns. A health marketing analysis and a series of year-long environmental assessments of surface and ground water were completed. The community was assisted with reaching consensus for future actions, emphasizing local control, enhanced county-based ownership, and sustainability of intervention efforts. OUTCOMES AND IMPLICATIONS: The Del Rio and East Tennessee State University partnership was instrumental in accomplishing its short-term objectives with the remediation of two major health issues. The more important long-term objectives of enhancing citizen leadership skills and developing a more action-oriented community infrastructure were also met. Using an experiential learning model, students practiced community organization skills, conflict resolution and problem-solving strategies. The campus-community partnership illustrated the advantages of experiential, multidisciplinary education and accentuated the positive aspects of collaborative planning and action. The partnership continues to provide expanded learning opportunities for students and contributes to the empowerment and self sufficiency of the community. The ripple effects of the model have become evident, with dramatic increases in university-wide efforts to increase partnership opportunities and enhanced support for service learning throughout the region. PMID- 14742083 TI - Quest for social accountability: experiences of a new health sciences university in Nepal. AB - CONTEXT: B.P. Koirala Institute of Health Sciences (BPKIHS), a new Health Sciences University in Nepal has taken several steps to respond to the societal needs and has adopted an integrated, partially problem based and community oriented curriculum. OBJECTIVE: The objective of this study was to measure the school's achievements in responding to societal needs. METHODOLOGY: A descriptive cross sectional questionnaire survey (N = 46) of the administrators, faculty, students/residents and the community. The questionnaire included statements on relevance, quality, cost effectiveness and equity in the education, service and research domains of a medical school. The data were analyzed by using WINKS 4.5, a statistical package for Windows. RESULTS: The responders satisfactorily rated BPKIHS. The mean rating (mean 3.11, SD = 1.06) was more than the satisfactory score (3). The responders were satisfied with the education (Mean = 3.26, SD = 1.06) and research (Mean = 3.12, SD = 1.10) but were less satisfied with the service domain (Mean = 2.94, SD = 0.98). The majority believed that the service is based on health care priorities (72%), and includes primary care (80%). A fair proportion felt the need for improvements in quality of care (50%) and cost effectiveness of care (46%). In general the faculty and administrators groups were more satisfied than the community and students/residents groups. CONCLUSION: The study was useful in identifying the school's strengths as well as weaknesses in responding to the societal needs. PMID- 14742085 TI - An interview of Rogayah Binti Jaafar. Interview by Jane Westberg. PMID- 14742084 TI - Students evaluation of a traditional and an innovative family medicine course in Saudi Arabia. AB - This study compares students' evaluation of a traditional and an innovative undergraduate family medicine (FM) courses. The old curriculum was traditional and teacher-centered. Changes in-line with the innovative learning concepts were introduced. While innovative course (IC) students had significant improvement in both their attitude towards innovative learning methods and self-assessment of knowledge, traditional course students had improvement only in self-assessment of knowledge. Students in both courses did not show post-cycle improvement in perception of their own skills and were dissatisfied with the Health Center (HC) tutors' training. The need to recruit trained family physicians at the affiliated HC became evident. IC students valued the exercise of adding their generated learning needs as part of the curriculum. Other lessons learned were presented. We hope that findings of this study would encourage medical colleges in the region to critically review their FM courses. PMID- 14742086 TI - An interview of Thomas Toltaku. Interview by Jane Westberg. PMID- 14742087 TI - Study design in qualitative research--1: Developing questions and assessing resource needs. AB - This is the second in a series of four papers on understanding and doing qualitative research [Frankel & Devers (2000) Qualitative research: a consumer's guide, Education for Health, 13, 113-123; Devers & Frankel (2000) Study design in qualitative research--2: sampling and data collection strategies, Education for Health, 13, 263-271]. Here, we focus on problems of study design, including question development, literature review, identifying a target audience and resource needs assessment. We provide a step-by-step description of major issues and choice points in the process. There are three key differences between qualitative and quantitative research designs. First, the logic of qualitative research is often inductive, rather than deductive, and consists of describing people's and groups' particular situations, meanings and experiences. Second, qualitative research designs are often emergent and flexible, and the research itself is quite dynamic. Third, the qualitative research process is non-linear and non-sequential. There is agreement that good qualitative studies answer clearly stated, important research questions. How qualitative research questions are formulated has implications for conducting a literature review. Some scholars believe that literature should be reviewed prior to beginning a study; others argue that this may impede the researcher from truly listening, observing and remaining open to new concepts and ideas. We offer suggestions about formulating research questions and how and when to conduct a literature review. Another important issue in conducting qualitative research is determining the resources that will be needed to conduct a study. These include internal resources, such as research skills, and external resources, such as personnel (expertise and time), equipment, supplies and travel. A description of typical resource and management issues in conducting a qualitative research study is included. PMID- 14742088 TI - Study design in qualitative research--2: Sampling and data collection strategies. AB - In two prior papers in our series on qualitative research [Frankel & Devers (2000a, 2000b) Qualitative research: a consumer's guide, Education for Health, 13, 113-123; Frankel & Devers (2000) Study design in qualitative research-1: developing research questions and assessing research needs, Education for Health, 13, 251-261], we examine two critical issues in qualitative research design: sampling, including identifying and negotiating access to research sites and subjects, and data collection and management. We describe these two key steps in the qualitative research design process, discuss challenges that often emerge when pursuing these steps, and provide guidelines for addressing them. Qualitative research most often uses "purposive," rather than random, sampling strategies. A good understanding of these sampling strategies and why they are used is central to designing a credible qualitative study. In addition, given the real-world context in which most qualitative research is carried out, identifying and negotiating access to research sites and subjects are critical parts of the process. We also provide suggestions for developing and maintaining productive and mutually satisfying research relationships with sites and subjects. Finally, data collection and management are often neglected subjects in qualitative research. We offer practical advice on how to collect and manage qualitative data, including factors to consider when deciding how structured the data collection process should be, the pros and cons of audio- and/or videotaping compared with note-taking, and tips for writing up field notes and document management. A forthcoming, final paper in the series will focus on qualitative data analysis and the publication of qualitative research results. PMID- 14742093 TI - Interaction of conjugated linoleic acid, sphingomyelin, and butyrate on formation of colonic aberrant crypt foci and immune functions in rats. AB - This study was designed to investigate possible additive or synergistic action among sphingomyelin (SPH), cis-9,trans-11-conjugated linoleic acid (CLA), and butyrate (BTY) against colon cancer and modulation of immune functions in vivo in male Sprague-Dawley rats. Each of the 5 groups of rats was fed either 35 mg SPH, 100 mg CLA, or 100 mg BTY/kg body weight, a combination of the 3 compounds at the same doses, or none of the compounds, for 7 wk. Rats were injected with azoxymethane, a colon carcinogen, to induce the formation of aberrant crypt foci, preneoplastic lesions of colon cancer. Parameters measured included number and multiplicity (number of crypts per focus) of aberrant crypts, immune functions such as innate immunity (natural killer cell cytotoxicity), humoral immunity (development of antibodies), and cell-mediated immunity (delayed-type hypersensitivity). Results show that the groups treated with SPH, CLA, and BTY individually had significantly higher natural killer cell (NK) cytotoxicity than the group treated with all compounds. The CLA group also had significantly higher NK activity than the control group. This study shows that the three compounds may not act additively or synergistically either to inhibit the development of aberrant crypts or to enhance immune functions. In fact, exposure to the combined compounds may be antagonistic to enhancement of NK function by the individual chemicals. PMID- 14742094 TI - Relationship between ambient air pollution and hospital admissions for cardiovascular diseases in kaohsiung, taiwan. AB - This study was undertaken to determine whether there is an association between air pollutants levels and increased number of hospital admissions for cardiovascular diseases (CVD) in Kaohsiung, Taiwan. Hospital admissions records for CVD and ambient air contaminant data collected from monitoring station in Kaohsiung were obtained for the period 1997-2000. The relative risk of hospital admission for CVD was estimated using a case-crossover approach, controlling for weather variables, day of the week, seasonality, and long-term time trends. In the single-pollutant model, on warm days (> or =25 degrees C) statistically significant positive associations were found between levels of particular matter of < 10 microm aerodynamic diameter (PM10), nitrogen dioxide (NO2), carbon monoxide (CO), and ozone (O3). On cool days (< 25 degrees C), all pollutants except O3 were significantly associated with increased CVD admissions. For the two-pollutant model, CO and O3 were both significant in combination with each of the other four contaminants on warm days. PM10 and NO2 remained significantly associated with elevated CVD admissions on warm days. On cool days, CO and NO2 remained statistically significant in all the two-pollutant models. This study provides evidence that higher levels of ambient contaminants, particularly CO, increase the risk of increased hospital admissions for CVD. PMID- 14742095 TI - Developmental immunotoxicity of lead in the rat: influence of maternal diet. AB - The effect of maternal dietary protein intake on lead-induced developmental immunotoxicity was studied in female Fischer 344 rats receiving lead acetate (250 ppm) or sodium acetate (control) in the drinking water during breeding and pregnancy until parturition. Dams were fed isocaloric diets (either 20% casein or 10% casein) from 2 wk prior to mating until the end of lactation. After weaning, dams and female offspring were given the 20% casein diet and regular water. Immune function was assessed in dams at 8 wk postpartum and in offspring at 13 wk of age. Dams showed no marked difference in any of the immune endpoints examined, regardless of diet or lead treatment. In contrast, lead exposure during early development produced a subsequent significant reduction of both the delayed-type hypersensitivity response and interferon gamma production in adult offspring independent of maternal diet. Lead-exposed offspring from the high-dietary protein group had significantly elevated production of both interleukin-4 and tumor necrosis factor alpha(TNF-alpha) with increased relative spleen weight and a decreased body weight compared to offspring in the lead control group. In contrast, lead-exposed offspring from dams receiving the low-protein diet had no marked change in TNF-alpha levels, relative spleen weight, or body weight, while interleukin-4 levels were significantly reduced compared with the lead control group. In conclusion, maternal dietary protein intake can modulate the immunotoxic effects of lead exposure during early development. This occurred at levels of protein intake and doses of lead exposure that produced no detectable effect on the maternal immune system. PMID- 14742096 TI - Quantitation of acrolein-protein adducts: potential biomarker of acrolein exposure. AB - Acrolein, an alpha,beta-unsaturated aldehyde, is a ubiquitous environmental toxic pollutant. Because of potential human exposure, there is a need for a sensitive, reliable, and specific method to monitor acrolein exposure. Acrolein is a potent electrophile and reacts with proteins mainly through Michael addition reaction, leading to acrolein-protein adducts (APA). The present study aimed to develop a competitive enzyme-linked immunosorbent assay (ELISA) method for the quantitation of APA in biological samples. Antibody to acrolein-keyhole limpet hemocyanin adduct was raised in rabbits, and the specificity of the antibody was determined by ELISA using acrolein-albumin adduct (AAA) or native albumin. A dose-dependent response was observed with AAA, but no immunoreactivity with native albumin. Further, lack of cross-reactivity of anti-acrolein antibody with formaldehyde-, malondialdehyde-, or 4-hydroxynonenal-albumin adducts indicates its specificity for acrolein. For the competitive ELISA, 1:16,000 diluted antisera was used with varying concentrations of AAA, which provided a linear detection range between 250 and 10,000 pg. To test the efficacy of the method for possible use as a biomarker of acrolein exposure, SD rats were orally administered 1 or 7 doses of 9.2 mg/kg/d acrolein. APA levels, quantitated in the serum, showed significantly greater formation (32% and 58% after 1 and 7 doses, respectively) in acrolein treated rats as compared to the controls. Western blot analyses of APA in the sera from acrolein-treated rats showed APA bands (especially 29, 31, and 100 kD) with greater intensity in comparison to controls, further supporting our ELISA results. These results suggest that quantitation of APA has potential to be used as biomarker of acrolein exposure and eventually for molecular dosimetry and risk assessment. PMID- 14742097 TI - A randomized, placebo-controlled, crossover study of ephedrine for SSRI-induced female sexual dysfunction. AB - The objective of this study was to determine whether ephedrine, an alpha- and beta-adrenergic agonist previously shown to enhance genital blood flow in women, has beneficial effects in reversing antidepressant-induced sexual dysfunction. Nineteen sexually dysfunctional women receiving either fluoxetine, sertraline, or paroxetine participated in an eight-week, double-blind, placebo-controlled, cross over study of the effects of ephedrine (50 mg) on self-report measures of sexual desire, arousal, orgasm, and sexual satisfaction. Although there were significant improvements relative to baseline in sexual desire and orgasm intensity/pleasure on 50 mg ephedrine 1-hr prior to sexual activity, significant improvements in these measures, as well as in sexual arousal and orgasmic ability also were noted with placebo. These findings highlight the importance of conducting placebo controlled trials for this condition. PMID- 14742099 TI - Sexual dysfunction in Parkinson's disease. AB - Sexual dysfunction is common in Parkinson's disease (PD). We investigated the premorbid and present sexual functioning of 75 people with PD (32 women and 43 men). Women reported difficulties with arousal (87.5%), with reaching orgasm (75.0%), with low sexual desire (46.9%), and wih sexual dissatisfaction (37.5%). Men reported erectile dysfunction (68.4%), sexual dissatisfaction (65.1%), premature ejaculation (40.6%), and difficulties reaching orgasm (39.5%). Premorbid sexual dysfunction may contribute to cessation of sexual activity during the course of the disease (among 23.3% men and 21.9% women). Associated illnesses, use of medications, and advanced stage of PD contributed to sexual dysfunction. PMID- 14742098 TI - Sexuality and health-related quality of life after prostate cancer in African American and white men treated for localized disease. AB - The purpose of this study was to examine differences in sexual attitudes and quality of life of White and African-American men who have undergone radical prostatectomy or radiation therapy for localized prostate cancer. Respondents included 1,112 White and 118 African-American men. Response rates differed by race, with 51% of White men and 28% of African-American men returning the questionnaire assessing demographics, medical history, sexual functioning, attitudes about seeking help for sexual problems, sexual self-schema, and health related quality of life. African Americans were more likely than Whites to have undergone radiation therapy (p <.0001) and were more likely to indicate that a desire to maintain sexual functioning influenced their treatment choice (p <.0001). African-American men also had more positive attitudes than did White men toward seeking help for sexual problems and were more likely to report seeking past help and intending to seek future help. African-American men reported more problems with sexual desire (p =.0003), although their sexual function scores did not differ significantly from those of Whites. African-American men may be more at risk for distress when prostate cancer treatment causes sexual dysfunction. PMID- 14742100 TI - A study of treatment choices in men with erectile dysfunction and reduced androgen levels. AB - A large cohort of men presenting with erectile dysfunction were offered concurrent measurement of serum total testosterone. Just over one sixth (17.7%) of the initial screening values of patients suggested the possibility of hypogonadism, however, when more rigorous estimation of serum testosterone and associated parameters and the presence of clinical symptoms were applied, 3% of the population had hypergonadotropic hypogonadism. Only 10 of 44 (22.7%) eligible men agreed to a trial of testosterone therapy, however, two thirds of this eligible group also required another erectogenic agent to resolve the erectile dysfunction. PMID- 14742107 TI - The Adolescent Prenatal Questionnaire: assessing psychosocial factors that influence transition to motherhood. AB - The purpose of the study was to develop a psychometrically reliable and valid instrument that assesses psychosocial factors that influence transition to motherhood in first-time pregnant adolescents 15-18 years of age. The descriptive, correlational study tested the Adolescent Prenatal Questionnaire (APQ) in 145 unmarried first-time pregnant adolescents 15-18 years of age who planned to keep their infants. Factor analysis yielded an interpretable four factor solution that accounted for 40% of the variance with adequate internal consistency estimates and test-retest correlations to merit further testing. PMID- 14742108 TI - Counseling about contraception among repeated aborters in Ho Chi Minh City, Vietnam. AB - We conducted qualitative interviews with 30 repeated abortion clients in Ho Chi Minh City, Vietnam. Only 5 women were practicing contraception regularly. A common reason for the nonusage of contraceptives was adverse health effects. Half of women were not counseled during the previous abortions, and contraceptives were given without explanation. Women pointed out that the current service was not enough or in-depth and was not meeting their needs. The type of counseling they wanted was a face-to-face discussion with a physician at a hospital using leaflets and pictures. Concerted efforts are needed to improve the quality of contraceptive counseling and to listen to women's voices. PMID- 14742109 TI - Birth outcomes across ethnic groups of women in Nepal. AB - This study was conducted at the Western Regional Hospital (WRH), Pokhara, Nepal. We investigated the relationship between selected socioeconomic variables, pregnancy indices, birth weight, and maternal health in women of different ethnic origins. In our cross-sectional data set, 29.8% of infants were born with a low birth weight. Our data analysis showed Indo-Aryan and lower caste ethnic groups had significantly lower weight babies than Tibeto-Burman and Newar groups. Further analysis showed that the Tibeto-Burman group received better care during pregnancy. Similarly, the Newar groups had significantly better nutritional intake than the Indo-Aryan and lower caste groups. The outcome of food restriction and antenatal care during pregnancy in specific ethnic groups has important implications for the health care delivery system. Health policymakers should target those ethnic groups and develop culturally based policies to reduce the incidence of low birth weight in Nepal. PMID- 14742110 TI - Lay advice on alcohol and tobacco during pregnancy. AB - Little is known about lay advice on prenatal alcohol and cigarette use and how this advice may complement or counteract advice from health professionals. In this study we examine the advice-giving role of female friends and relatives ("confidantes"). Survey data were collected from 105 low-income pregnant women about prenatal alcohol and cigarette use and confidante advice about these behaviors. Focus groups with 9 confidantes of pregnant smokers examined the advice they offered about substance use, their perceptions of these behaviors, and their roles as advice-givers. The rate of prenatal drinking was low among pregnant smokers, consistent with confidantes' high risk perceptions and zero tolerance for drinking, but low risk perceptions and high tolerance for smoking. Confidantes described barriers to providing advice about smoking cessation. Because confidantes perceived their role to be distinct from, and in some ways more influential than, that of doctors their advice should be considered in the development of prenatal substance use interventions. PMID- 14742111 TI - Women's postnatal experience following a medically complicated pregnancy. AB - Within the social science literature on reproduction, relatively little attention has been paid to the areas of major illness during pregnancy and postnatal physical health. In this paper I present the results of a qualitative study of the postnatal health experiences of 15 women who had suffered a serious illness during pregnancy, the onset of which either predated or developed during pregnancy. Recently delivered women with continuing health problems face additional difficulties at a time when they are recovering from the physical and psychological processes of birth and are adjusting to the care of the new baby or grieving a loss. A few women in this study had experienced health problems that were resolved at or soon after the birth. Those with continuing or chronic illness described attempts to regain normality that involved both restoring bodily health and reestablishing their control over symptoms of the illness. Professional help with this process was often reported to be limited. PMID- 14742112 TI - Protective strength factors, resources, and risks in relation to depressive symptoms among childbearing women of Mexican descent. AB - As an immigrant population becomes more prominent in an established culture, research strategies for examining intragroup differences are needed to understand their health care risks and strengths. However, acculturation also must be taken into account. Our objective in this secondary analysis was to identify the most useful acculturation parameter for examining depressive symptoms in relation to strengths and resources among women of Mexican descent living in the United States during the vulnerable perinatal period and to examine other intragroup differences among the women by childbearing status (pregnant or postpartum). Our secondary analysis was done with a subsample of 129 women from a larger cross sectional study of 315 women of Mexican descent who were recruited from three urban community health centers and an associated bilingual school in the United States. Exposure to the United States in childhood, a poor sense of mastery, and dissatisfaction with life were more related to depressive symptom experience than childbearing status or more traditional demographic variables such as age, income, or education. PMID- 14742113 TI - Kathleen Mahoney's lucid presentation at the ICOWHI conference in June. PMID- 14742115 TI - Women's somatic styles: rethinking breast self-examination education. AB - In this ethnographic study I explored women's somatic and sexual experiences, reception of breast self-examination (BSE) messages, and reactions to the practice of BSE. Mainstream BSE education uses messages that deemphasize the woman, her breasts, and her relationship to them as sexual. The turbid confluence of societally eroticized breasts and self-touch taboos makes it unlikely that women filter these messages in an asexual way. Using grounded theory, I examined women's expression of the self-body relationship and the sociocultural milieu within which women consider BSE education materials. Seven subjects varying in age, sexual orientation, parenting, and relationship status were interviewed about their physical experiences, self-touch, and body image. Subjects also participated in a BSE class and focus group, and composed a journal entry describing their reactions to practicing BSE. Discernible patterns in somatic memories, somatic styles, and reactions to BSE educational messages were found. This study suggests a need for a more consciously feminist approach to BSE education. PMID- 14742116 TI - Breast cancer screening methods: a review of the evidence. AB - Breast cancer remains a major health problem among Canadian women. Efforts directed at primary prevention of the disease are limited. Secondary prevention through screening appears to be the most promising intervention available in controlling the disease. In recent years, there have been ongoing debates over the effectiveness of available breast screening modalities (breast self examination [BSE], clinical breast examination [CBE], and screening mammography). In this article I provide an overview of evidence related to each of the three breast screening modalities. The evidence shows that screening mammography and proper examination of breasts can be useful in reducing breast cancer mortality. PMID- 14742117 TI - Maintaining respectability and responsibility: gendered labor patterns among women injection drug users. AB - Gender-related factors and the social and economic conditions that impact the lives and health of women injection drug users (IDUs) in Chicago are described. Although study participants are highly imaginative and resourceful in terms of income-generating self-sufficiency, they engage in a variety of behaviors that put them at risk of contracting infectious diseases such as HIV and hepatitis B and C viruses. We point out that labor inequalities experienced by women IDUs, together with the gender ideologies that support those inequalities, provide reduced opportunities to practice harm reduction and other health care options. PMID- 14742118 TI - Behavioral practices and mortality in women former college athletes and nonathletes. AB - In 1981-1982 we first studied the long-term health of women college graduates, college athletes, and a comparison group of nonathletes; the women's average age was 39 years. In a 15-year follow-up we investigated changes in health, behavioral practices, and mortality. Smoking was a highly significant risk factor for mortality: The hazard ratio was 2.2. Other behavioral risk factors were ever smoking and not currently engaging in regular exercise. Behavioral practices that include not smoking, doing regular exercise, and consuming moderate amounts of alcohol may lead to lower mortality. PMID- 14742120 TI - Perceived effects of testosterone replacement therapy in perimenopausal and postmenopausal women: an internet pilot study. AB - Women experience somatic, mood, energy, and libido changes during menopause. Testosterone replacement therapy is being prescribed in conjunction with estrogen replacement therapy (ERT) for these symptoms. Little is known about women's perceptions of the benefits and risks of testosterone, so an Internet survey was conducted to elicit women's perceptions of the effects on mood, energy, sexuality, and somatic symptoms. A sample of 61 women primarily from the United States and Canada participated in the survey; one woman from Germany and one from Sweden also took part in the survey. Participants reported significantly improved sexuality and improved mood and vigor, with a decline in fatigue. Of the 7 somatic complaints assessed, improvements were noted in 5 (sleep disturbances, hot flashes, night sweats, vaginal dryness, and appetite). A prospective clinical trial is needed to confirm these results. PMID- 14742119 TI - Medicalized menopause, US style. AB - The construction of menopause as an estrogen deficiency condition originated primarily in the United States and is being exported to many other countries throughout the world. I have identified specific cultural assumptions and policies in the United States as well as the ubiquitous gender biases that have contributed to the medicalization of menopause. Since medicalized menopause is causing harm to women worldwide, it is imperative that it be addressed by the international community. PMID- 14742125 TI - None of us will ever be the same again: reactions of American midlife women to 9/11. AB - According to terror management theory (TMT), an event that heightens awareness of death produces the need to defend against existential anxiety. The horrifying events of September 11, 2001 (9/11), created an unparalleled opportunity to apply TMT beyond the laboratory. This study examined post-9/11 stress (via perceived stress scale [PSS] scores) and interview responses of a diverse community sample of American midlife women (ages 35-60). Previous studies showed that many women have high stress during midlife, suggesting that 9/11 could have a unique impact on this segment of the U.S. population. Education of the sample ranged from 12 to 23 years. Seventy-five percent had children and 70.6% were married. Data analysis showed that 4 to 6 months after 9/11, 61% of the women were still distressed, exhibiting symptoms of fear, sadness, anger, powerlessness, distrust, and vigilance. Highly stressed women (upper 25% on PSS) differed in several respects from low-stress women (lower 25% on PSS). As predicted by TMT, core values central to a woman's world view were activated by 9/11. Patriotism and altruism increased, but bigotry intensified as well. Major changes (e.g., marriage, moving) were undertaken only by a small percentage (18%), but all expressed the view, "None of us will ever be the same again." Women who had experienced previous trauma felt that their background actually helped them cope. PMID- 14742126 TI - Taking a different path: menstrual preparation for adolescent girls living apart from their mothers. AB - Menarche is a significant yet anxiety producing event. Preparation for menarche combats this anxiety. When adolescent girls reach menarche they often seek information about menstruation from their mothers. Yet not every girl lives with her mother. The purpose of my grounded theory study is to explore the menarcheal experience of 16 girls living without their mothers. The findings indicate that prior to menarche participants experienced similarities to girls who lived with their mothers. Differences became apparent at menarche when participants had to wait for an available female to answer questions, did not have a female available to ask, or felt embarrassed discussing menstruation with their fathers. Providers need to be sensitive to menarcheal preparation issues for girls living without their mothers. PMID- 14742127 TI - Promoting women's health-seeking behavior: research and the empowerment of women. AB - Despite advances in medical knowledge, commentators agree that the greatest gains in health will come through behavioral change. Women must change their health seeking behavior; worldwide, health advocates find that even though services may be provided for women, it does not guarantee that women use them. The purpose of this article is to help researchers, as women's advocates, understand why. Specifically, we present a tool that helps identify barriers to, as well as facilitators of, women's health seeking. Unlike conventional approaches that focus on psychological or personal facilitators of health seeking, we use a method that locates the individual within her sociocultural context. Such an approach helps us differentiate women's practical needs for health care from their strategic interest in gender equity; in doing so, we advance a distinctly feminist approach to women's health promotion. PMID- 14742128 TI - The quality of women's life after mastectomy in Poland. AB - From the increasing number of women who recovered from breast cancer arises more academic interest in their subjective opinions about the quality of their life (QL), particularly the physical, psychological, and social aspects of it. Our aim in this study was to evaluate the QL of Polish women after mastectomy. The research was performed on 480 randomly chosen patients who had been subjected to surgical treatment. The functional assessment of cancer therapy (FACT) scale version 4 and hospital anxiety and depression scale (HADS) were used during the research procedure. The lowest results within the global opinion of QL were obtained within the emotional sphere. The most frequent mental states experienced by the examined patients were worry, sadness, nervousness, fear, and depression. PMID- 14742129 TI - Missing girls and son preference in rural India: looking beyond popular myth. AB - Survival chances of girls in parts of South and East Asia have been adverse. Female foeticide, infanticide, abandonment, out-adoption, under-reporting of female births, and selective neglect of girls leading to higher death rates, have contributed to this adversity. Here, I reflect on an observed skewed sex ratio at birth in rural West Bengal, and discuss female foeticide, infanticide, son preference, and abortion. More boys were recorded at birth, and the majority of women desired sons. Trajectories of selective neglect of and discrimination against daughters have been researched extensively, but the related issues of female foeticide and infanticide have been less examined. PMID- 14742130 TI - Observations and advertising: controversies in the prescribing of hormone replacement therapy. AB - Since scientists in the 1930s discovered sex hormones, hormone replacement therapy (HRT) has been a source of considerable controversy and debate. This debate continues with the publication of the results of the Women's Health Initiative, a large randomized controlled trial that has once again altered the way women and their care providers view this therapy. Over the years the evidence increasingly has suggested that HRT is not effective for the prevention of cardiovascular disease in women. However, the prevailing opinion denied this, and millions of women were prescribed estrogen with or without progestin under the assumption that cardioprotection was possible. Why did this practice continue in the face of mounting evidence of harm? In this article we will explore the role that the media and pharmaceutical companies have played in the widespread use of these hormones despite evidence of significant side effects and lack of prevention efficacy. PMID- 14742131 TI - Predictors of women's Internet access and Internet health seeking. AB - A random, population-based sample of 431 women aged 18-74 in King County, Washington, USA, completed a survey module on Internet use and access. Level of mental health, level of general health perceptions, older age, and higher income predicted women's health-related Internet use. Participants without access reported various barriers to obtaining access; perceived lack of usefulness of the Internet as an information source and unfamiliarity with using this technology appear equally important reasons as financial cost for not adopting the Internet. Internet use motivators are complex; these findings have relevance to the design of Internet-based interventions. PMID- 14742134 TI - Effects of renal diseases on the regulation and expression of renal and hepatic drug-metabolizing enzymes: a review. AB - 1. The activity of drug-metabolizing enzymes (DMEs) in extrahepatic organs is highest in the kidneys. Generally, the kidneys contain most, if not all, of the DMEs found in the liver. Surprisingly, some of these DMEs show higher activity in the kidneys than in the liver. 2. Most of the renal DMEs are localized in the cortex of the kidneys, especially in the proximal tubules. DMEs are also found in the distal tubules and collecting ducts. 3. Renal diseases such as acute and chronic renal failure and renal cell carcinoma alter the regulation of both hepatic and extrahepatic phase I and II DMEs. Changes in the expression of these DMEs seem to be tissue and species specific. 4. Generally, there is significant down-regulation of most of the phase I and a few of phase II DMEs at the protein, mRNA and activity levels. Unfortunately, the mechanisms leading to the alteration in DMEs in renal diseases remain unclear, although many theories have been made. 5. The presence of some circulating factors such as cytokines, nitric oxide, parathyroid hormones and increased intracellular calcium play a role in the regulation of DMEs in renal diseases. PMID- 14742135 TI - Comparison of the metabolism of ethylene glycol and glycolic acid in vitro by precision-cut tissue slices from female rat, rabbit and human liver. AB - 1. The metabolism of [1,2-(14)C]-ethylene glycol and [1,2-(14)C]-glycolic acid was studied in vitro using precision-cut tissue slices prepared from the livers of female Sprague-Dawley rats, New Zealand white rabbits and humans. The time course for production of metabolites formed from ethylene glycol at concentrations from 3 to 40 mM was determined to compare quantitatively the differences between species in the rates and amounts of formation of glycolic acid, the presumed developmental toxicant of ethylene glycol. The rates of metabolism of glycolic acid to glyoxylic acid at concentrations from 0.05 to 16 mM by liver tissue from the different species were also determined. The apparent V(max)/K(m) for the metabolic conversions of ethylene glycol to glycolic acid and for glycolic acid to glyoxylic acid in liver tissue from the different species were obtained. 2. There were qualitative differences in the metabolic profiles and quantitative differences in the formation of glycolic acid between the mammalian liver systems. There was an average of 10-fold less glycolic acid produced by liver slices from rabbits compared with rats. With the human liver, the formation of glycolic acid was not detectable using tissue from three of four human donors. A low level of glycolic acid was detected in one liver slice incubation from one of the four subjects, but only at one extended time point; glyoxylate was detected with liver slices from all four humans. 3. Liver slices prepared from female Sprague-Dawley rats, female New Zealand White rabbits and three female human subjects all metabolized glycolic acid to glyoxylic acid. Human liver tissue was the most effective at further metabolizing glycolic acid to glyoxylic acid. The ratios of V(max)/K(m), representing the relative clearance of glycolic acid from liver tissue, were approximately 14:9:1 for human, rat and rabbit liver, respectively. 4. Precision-cut liver slices maintained in dynamic organ culture are good predictors of metabolism by liver tissue in vivo. The results of the present study therefore indicate that levels of glycolic acid, if formed in vivo, following exposures to similar concentrations of ethylene glycol, would be lower in humans than in rabbits and rats. PMID- 14742136 TI - Optimizing bacterial expression of catalytically active human cytochromes P450: comparison of CYP2C8 and CYP2C9. AB - 1. Methods for the co-expression in Escherichia coli of human cytochrome P450 (CYP) 2C8 and CYP2C9 with NADPH-cytochrome P450 reductase (OxR) to produce a catalytically active system were compared. 2. Approaches assessed were expression of a CYP:OxR fusion construct, bicistronic plasmids, simultaneous transformation with CYP and OxR plasmids, and separate expression of CYP and OxR with reconstitution of activity by mixing the bacterial membranes. Two N-terminal modifications (Delta3-20 and 17alpha-leader) of the individual P450s were additionally investigated. 3. Each approach gave efficient expression of CYP2C8 and CYP2C9, but the bicistronic constructs under the expression conditions used gave low OxR expression and low catalytic activity. CYP expression was higher with the Delta3-20 construct for CYP2C9 and with the 17alpha-presequence construct for CYP2C8. 4. Using torsemide as substrate, all methods gave catalytically active systems with K(m) values similar to human liver microsomes. Mixing bacterial membranes containing separately expressed CYP and OxR reconstituted a catalytically active system with the Delta3-20 construct for CYP2C9 but not for CYP2C8, and with neither of the 17alpha- presequence constructs. OxR co-expressed with CYP in the same membrane interacted with CYP to reconstitute activity more effectively than addition of exogenous OxR membranes. 5. Expression construct and OxR co-expression strategy should be individualized for CYP isoforms. PMID- 14742137 TI - Oxidative one-carbon cleavage of the octyl side chain of olanexidine, a novel antimicrobial agent, in dog liver microsomes. AB - 1. The oxidative one-carbon cleavage reaction in the octyl side chain of olanexidine [1-(3,4-dichlorobenzyl)-5-octylbiguanide], a new potent biguanide antiseptic, was characterized in dog liver microsomes. 2. Olanexidine was initially biotransformed to a monohydroxylated metabolite, 8-[5-(3,4 dichlorobenzyl)-1-biguanidino]-2-octanol (DM-215), and DM-215 was subsequently oxidized to the diol derivative, 8-[5-(3,4-dichlorobenzyl)-1-biguanidino]-1,2 octandiol (DM-220). DM-220 was further biotransformed to 2-hydroxy aldehyde derivative, 2-hydroxy carboxylic acid derivative, and an oxidative C-1-C-2 bond cleavage metabolite, 7-[5-(3,4-dichlorobenzyl)-1-biguanidino] heptanoic acid [DM 223 (C7), a seven-carbon chain derivative], after incubation with dog liver microsomes. 3. DM-223 formation required NADPH as a cofactor and was inhibited by quinidine and quinine, relatively selective inhibitors of CYP2D subfamilies in dogs. 4. The results suggest that the one-carbon fragment of the octyl side chain of olanexidine could be removed by the oxidative C-C bond cleavage with the possible involvement of cytochrome P450 systems such as CYP2D subfamily. This oxidative C-C bond cleavage reaction by cytochrome P450s could play an important role in the removal of one-carbon fragment of other drugs or endogenous compounds containing aliphatic chains. PMID- 14742138 TI - High-throughput screening for the assessment of time-dependent inhibitions of new drug candidates on recombinant CYP2D6 and CYP3A4 using a single concentration method. AB - 1. The inhibitory effects of various test compounds on recombinant human CYP3A4 activity assayed by fluorescent metabolite formation from 7-benzyloxyquinoline (7 BQ) and the effect of pre-incubation on inhibition were evaluated using the microtitre plate assay with multiple concentrations of test compounds (multiple concentration method). 2. Among the test compounds studied, ketoconazole inhibited CYP3A4 activity most extensively, followed by miconazole, troleandomycin, terfenazine and midazolam. The IC(50) values of other compounds exceeded 10 microM, but those of many compounds decreased after pre-incubation. The inhibitory effects of verapamil, amiodarone and diltiazem after pre incubation were 205, 154 and 833 times greater than those in the case of co incubation, respectively. 3. To assess the inhibitory effects more readily, the validity of the microtitre plate assay with a single concentration of the test compound (single concentration method) was studied. The accuracy of the automated dispensation and the coefficient of variation on enzyme activity were approximately 3%. 4. The IC(50) values estimated using the per cent of residual activity from the single concentration method matched closely those from the multiple concentration method. When the IC(50) value as inhibitor concentration was used for a single concentration method, the method enabled easy estimation of inhibitory patterns (such as competitive or time-dependent inhibition) on cytochromes P450. Therefore, from the ease of the technique, automation of the microtitre plate assay and application of the single concentration method might be useful for inhibitory assessment of cytochromes P450 more than that of current conventional methods. PMID- 14742139 TI - Disposition of a novel and potent alpha(v)beta3 antagonist in animals, and extrapolation to man. AB - 1. The disposition of 3-[2-oxo-3-[3-(5,6,7,8-tetrahydro-[1,8]naphthyridin-2-yl) propyl]-imidazolidin-1-yl]-3(S)-(6-methoxy-pyridin-3-yl)propionic acid (compound A), a potent and selective alpha(v)beta(3) antagonist, was characterized in several animal species in support of its selection for preclinical safety studies and potential clinical development. 2. Compound A exhibited marked species differences in pharmacokinetics; the plasma clearances and bioavailabilities ranged from 33-47 ml min(-1) kg(-1) in rats and mice to 4-9 ml min(-1) kg(-1) in dogs and monkeys, and about 20% in rats to 70-80% in dogs and monkeys, respectively. Both the intravenous (i.v.) and oral kinetics of compound A were linear over the dose range studied in dogs (0.1-5 mg kg(-1) i.v. and 0.25-20 mg kg(-1) orally [p.o.]) and rats (1-30 mg kg(-1) i.v. and 4-160 mg kg(-1) p.o.). 3. Compound A was eliminated substantially by urinary excretion; the urinary recovery of the unchanged drug was 67% in rhesus, 48% in dogs and about 30% in rats. In these animal species, biotransformation was modest. 4. Following i.v. administration of [(14)C]-compound A to rats, the radioactivity rapidly distributed to all tissues investigated, with high levels of the radioactivity detected in liver, kidney and intestine soon after the drug administration. The radioactivity declined rapidly, with less than 1% of the i.v. dose remaining at 30-h post-dose. 5. Compound A was moderately bound to plasma proteins, with unbound fractions of 26, 20, 14 and 5% for rats, dogs, monkeys and humans, respectively. It was bound primarily to human alpha(1)-acid glycoprotein (about 85% binding at 0.1% concentration), as compared with human albumin (< 50% binding at 4% concentration). 6. Using simple allometry, compound A was predicted to exhibit relatively low clearance (1-3 ml min(-1) kg(-1)) and low volume of distribution (0.1-0.3 l kg(-1)) in humans. Based on the predicted values, compound A was projected to exhibit a favourable oral pharmacokinetic profile in humans, with good bioavailability (50-80%). These predicted values provided a basis for compound selection for further development. PMID- 14742140 TI - Altered expression of sulfotransferases, glucuronosyltransferases and mrp transporters in FVB/mrp1-/- mice. AB - 1. Genetically altered mice increasingly are being used in toxicology and pharmaceutical development. As such, knowledge of the compensatory activity of enzymes is critical when interpreting the results of studies using these animals. 2. The present study examined alterations in hepatic phase I and II enzyme activity, and alterations in phase III (transporter) RNA expression, between FVB mice and mice lacking the multidrug resistance-associated protein 1 (mrp1) gene (FVB/mrp1-/- mice). It was hypothesized that other transporters and phase I and II enzymes would be increased in the FVB/mrp1-/- mice, presumably as a compensatory mechanism. 3. No differences was found in hepatic cytochrome P450 activity between FVB and FVB/mrp1-/- mice, nor were there differences in the amount of total hepatic glutathione or in glutathione S-transferase enzyme activity. 4. However, sulfotransferase activity towards 2-naphthol was significantly increased by 2.6-fold in the FVB/mrp1-/- mice, whereas glucuronosyltransferase activity towards both 4-nitrophenol and testosterone was significantly reduced 1.5-fold. In addition, mrp2 RNA expression was significantly increased by 3.4-fold and mrp5 expression was significantly increased by 1.6-fold in the FVB/mrp1-/- mice. 5. Mice lacking mrp1 have significantly increased hepatic transcription of at least two other ATP-binding cassette transporters, as well as increased 2-naphthol sulfotransferase activity, presumably to compensate for the lack of mrp1. PMID- 14742141 TI - Hepatic metabolism of diallyl disulphide in rat and man. AB - 1. The metabolism of diallyl disulphide was investigated in vitro with rat and human liver cell subfractions and ex vivo with an isolated perfused rat liver. 2. Diallyl disulphide was oxidized to diallylthiosulphinate by rat liver microsomes with an apparent K(m) = 0.86 +/- 0.1 mM and an apparent V(max) = 0.47 +/- 0.12 nmol min(-1) mg(-1) protein (mean +/- SE). Both cytochrome P450 (CYP) and flavin containing monooxygenases were involved, with CYP2B1/2 and CYP2E1 being the most active CYP enzymes. 3. In rat and man, microsomal oxidation of allylmethyl sulphide to allylmethyl sulphoxide and allylmethyl sulphone also occurred, although at a low rate. Diallyl disulphide was also metabolized to allylglutathione sulphide and allylmercaptan. In addition, diallylthiosulphinate reacted non-enzymatically with glutathione to form allylglutathione sulphide. 4. When an isolated rat liver was perfused with diallyl disulphide, the metabolites allyl mercaptan, allylmethyl sulphide, allylmethyl sulphoxide, allylmethyl sulphone and allylglutathione sulphide were detected primarily within the liver tissue, with only small amounts of metabolites found in the bile and perfusion medium. The pharmacokinetic parameters for diallyl disulphide were t(1/2) = 6.09 min; AUC(0- infinity ) = 4.77 min mmol l(-1); clearance = 34.22 ml min(-1). 5. A scheme for the metabolism of diallyl disulphide in rat and man is proposed. PMID- 14742142 TI - Identification of the human cytochrome P450s responsible for the in vitro metabolism of a leukotriene B4 receptor antagonist, CP-195,543. AB - 1. The major human cytochrome P450 (CYP) form(s) responsible for the metabolism of CP-195,543, a potent leukotriene B4 antagonist, were investigated. 2. Incubation of CP-195,543 with human liver microsomes resulted in the formation of three major metabolites, M1-3. M1 and M2 were diastereoisomers and formed by oxidation on the benzylic position. M3 was formed by aromatic oxidation of the benzyl group attached to the 3-position of the benzopyran ring. 3. The results from experiments with recombinant CYPs, correlation studies and inhibition studies with form-selective inhibitors and a CYP3A antibody strongly suggest that the CYP3A4 plays a major role in the metabolism of CP-195,543. Recombinant CYP3A5 did not metabolize CP-195,543. 4. The apparent K(m) and V(max) for the formation of M1-3 in human liver microsomes were determined as 36 microM and 4.1 pmol min( 1) pmol(-1) P450, 44 microM and 10 pmol min(-1) pmol(-1) P450, and 34 microM and 2.0 pmol min(-1) pmol(-1) P450, respectively. The average in vitro intrinsic clearance for M2 was the highest both in human liver microsomes and recombinant CYP3A4 compared with M1 and M3. Intrinsic clearance for M2 in human liver microsomes and recombinant CYP3A4 was 0.231 and 0.736 ml min(-1) pmol(-1) P450, respectively. The intrinsic clearances for M1 and M3 in human liver microsomes and CYP3A4 were 0.114 and 0.060 and 0.197 and 0.088 ml min(-1) pmol(-1) P450, respectively. This suggests that benzylic oxidation is the predominant phase I metabolic pathway of CP-195,543 in man. PMID- 14742143 TI - Inhibition of rat liver sulfotransferases SULT1A1 and SULT2A1 and glucuronosyltransferase by dietary flavonoids. AB - 1. Dietary flavonoids including kaempferol, quercetin, genistein and daidzein were tested for their ability to alter the conjugation of oestradiol (E(2)) via rat liver sulfotransferases and glucuronosyltransferase. 2. All four flavonoids inhibited the sulfonation of E(2) via phenol sulfotransferase, SULT1A1 with IC(50)s ranging from 0.29 to 4.61 micro M. Sulfonation of dehydroisoandrosterone (DHEA) via hydroxysteroid sulfotransferase, SULT2A1, was inhibited by higher amounts of the flavonoids (IC(50)s ranging from 34 to 116 micro M). 3. All flavonoids inhibited the formation of E(2)-beta-glucuronides (at carbon atoms 3 and 17) with IC(50)s ranging from 43 to 260 micro M. Glucuronidation of 4 methylumbelliferone (4-MU) was inhibited by high amounts of the flavonoids (IC(50)s ranging from 860 to 1550 micro M). 4. Hydrolysis of sulfonated oestrogens via arylsulfatase-c (ARSC) or 4-methylumbelliferone beta-glucuronidate (MUG) were not inhibited by the flavonoids. 5. It is concluded that SULT1A1 but not SULT2A1 or glucuronosyltransferase is highly sensitive to inhibition by dietary flavonoids. The potency of the inhibition for SULT1A1 (quercetin > kaempferol > genistein > daidzein) suggests a dependency on the number and position of hydroxyl radicals in the flavonoid molecule. PMID- 14742144 TI - S-oxidation of S-methyl-esonarimod by flavin-containing monooxygenases in human liver microsomes. AB - 1. Studies using human liver microsomes and recombinant human cytochrome P450 (CYP) and flavin-containing monooxygenase (FMO) were performed to identify the enzymes responsible for the metabolism of S-methyl-esonarimod (M2), an active metabolite of esonarimod (KE-298, a novel antirheumatic drug). 2. S-oxidative activities of M2 significantly correlated with those of methyl p-tolyl sulfide, a specific substrate of FMOs, as tested using 10 different human liver microsomes (r(2) = 0.539, p<0.05). Thermal treatment of microsomes reduced the S-oxidative activity in the absence of the NADPH-generating system at 45 degrees C for 5 min. However, methimazole, a known competitive substrate of FMOs, was a weak inhibitor of the S-oxidation in liver microsomes. 3. Recombinant human FMO1 and FMO5 produced M3 in greater quantities than recombinant human FMO3. The S-oxidation of M2 by recombinant human FMO5 was not appreciably inhibited in the presence of methimazole. In contrast, methimazole was effective in suppressing the catalytic activity of recombinant human FMO1 and FMO3. 4. The apparent K(m) (K(m app)) for the S-oxidation of M2 in human recombinant FMO5 (2.71 microM) was similar to that obtained using human liver microsomes (2.43 microM). 5. The present results suggest that the S-oxidation of S-methyl esonarimod reflects FMO5 activity in the human liver because the recombinant FMO5 data match well with the human liver microsomal experiments. PMID- 14742145 TI - 'Open access' generic method for continuous determination of major human CYP450 probe substrates/metabolites and its application in drug metabolism studies. AB - 1. An 'open access' generic high-performance liquid chromatography method was developed for different combination sets each containing specific cytochrome P450 probe substrate and the corresponding metabolite. Method development, optimization and validation were carried out with the following combinations: phenacetin + paracetamol + internal standard (IS, celecoxib), bufuralol + hydroxybufuralol + IS, testosterone + 6beta-hydroxytestosterone + IS, chlorzoxazone + 6-hydroxychlorzoxazone + IS, coumarin + 7-hydroxycoumarin + IS, tolbutamide + hydroxytolbutamide + IS, and diazepam + desmethyldiazepam + IS. 2. The assay procedure involved a simple one-step liquid/liquid extraction followed by reverse phase chromatography (Inertsil ODS 3V column) employing a ternary gradient system and the eluate was monitored by a photodiode array/fluorescence detector. The standard curve for each compound, in the concentration range 0.1-10 microg ml(-1), in various sets was linear (r(2)>0.99) and absolute recoveries of all analytes were >90%. The lower limit of quantification was 0.1 microg ml(-1). The intraday precision and accuracy in the measurements of quality control were <15% relative standard deviation and <15% deviation from nominal values, respectively. 3. Each combination set was tested with individual chemical inhibitors (furafylline, quinidine, ketoconazole, disulfiram, diethyldithiocarbamate, sulphaphenazole and tranylcypromine) and all analytes were well resolved. Overall, the assay is simple, uses conventional instrumentation and provides a scope to analyse all cytochrome P450 combination sets continuously. The application of the method in the cytochrome P450 liability screen of novel compounds is also presented. PMID- 14742146 TI - Pharmacokinetics and metabolism of NO-1886, a lipoprotein lipase-promoting agent, in cynomolgus monkey. AB - 1. The study was conducted to investigate the pharmacokinetics and metabolism of NO-1886 (diethyl 4-[(4-bromo-2-cyanophenyl) carbamoyl] benzylphosphonate) in cynomolgus monkeys. 2. After single intravenous administration of NO-1886 at a dose of 3 mg kg(-1), the total clearance (CL(tot)), area under the plasma concentration-time curve (AUC(0-)(t)), half-life (t(1/2)), and volume of distribution (V(d)) in cynomolgus monkeys were 531 ml h(-1) kg(-1), 5.63 micro g h ml(-1), 0.96 h and 679 ml kg(-1), respectively. The AUC(0-)(t) for oral administration of NO-1886 (3 mg kg(-1)) was 4.23 micro g h ml(-1) and the bioavailability was 75%. 3. M-2 (ethyl 4-[(4-bromo-2-cyanophenyl) carbamoyl] benzylphosphonate) and M-3 (4-[(diethoxy-phosphoryl) methyl)] benzoic acid) were present as metabolites in plasma and urine. In faeces, M-2 was present but M-3 was not. 4. The major metabolite of NO-1886 in liver S9 or microsomes was M-2 in the presence of NADPH. On the other hand, M-3 was formed in the absence of NADPH in liver S9 or microsomes and its formation was inhibited by bis-( p-nitrophenyl) phosphate (BNPP) in liver S9, suggesting that the formation of M-3 was catalysed by carboxylesterase. 5. The findings suggest that the main metabolic pathway of NO-1886 in cynomolgus monkeys is the O-deethylation of NO-1886 to M-2, as in rats and humans, and that the hydrolysis of the amide bond is a minor metabolic pathway. PMID- 14742148 TI - Working memory deficits after traumatic brain injury: catecholaminergic mechanisms and prospects for treatment -- a review. AB - PRIMARY OBJECTIVE: To review the neural circuitry and neurochemistry of working memory and outline the evidence for working memory deficits after traumatic brain injury, and the evidence for the use of catecholaminergic agents in the amelioration of these deficits. Current knowledge gaps and research needs are identified. MAIN OUTCOMES AND RESULTS: Impairments in working memory are a core component of the cognitive deficits associated with traumatic brain injury. Recent progress in understanding the neural circuitry and neurochemistry of working memory suggests that catecholamines play a central role in the activation and regulation of working memory and thus lays a framework in which to consider the use of catecholaminergic agents (dopaminergic and alpha-2 adrenergic agonists) in the treatment of specific cognitive deficits after traumatic brain injury. CONCLUSIONS: The combined methods of cognitive neuroscience, functional brain imaging and neuropharmacology are proposed as an excellent method for studying working memory deficits. A strong rationale exists for the targeted use of catecholaminergic agonists in the treatment of working memory deficits after traumatic brain injury. PMID- 14742149 TI - Natural history of chronic subdural haematoma. AB - This review will clarify the natural history of chronic subdural haematoma (SDH). Chronic SDH has dual origins, one from subdural hygromas (SDG) and the other from acute SDHs. It occurs only in patients with a suitable pre-morbid condition, i.e. sufficient potential subdural space (PSS). In unresolved SDGs, proliferation of dural border cells produces the neomembrane. Unresolved SDGs become chronic SDHs by repeated micro-haemorrhages from fragile new vessels, which were grown into the neomembrane. When PSS is sufficient, acute SDHs may become chronic SDHs. Chronic SDHs enlarge when rebleeding exceeds absorption and they become symptomatic. When the neomembrane is matured, the neocapillary is no longer fragile. If absorption exceeds rebleeding, the haematoma will disappear. Maturation of the neomembrane and stabilization of the neovasculature eventually result in spontaneous resolution. The fate of chronic SDH depends on the pre morbid status, the dynamics of absorption-expansion and maturation of the neomembrane. PMID- 14742150 TI - Clinical progression and outcome of dysphagia following paediatric traumatic brain injury: a prospective study. AB - OBJECTIVES: To provide a preliminary clinical profile of the resolution and outcomes of oral-motor impairment and swallowing function in a group of paediatric dysphagia patients post-traumatic brain injury (TBI). To document the level of cognitive impairment parallel to the return to oral intake, and to investigate the correlation between the resolution of impaired swallow function versus the resolution of oral-motor impairment and cognitive impairment. PARTICIPANTS: Thirteen children admitted to an acute care setting for TBI. MAIN OUTCOME MEASURES: A series of oral-motor (Verbal Motor Production Assessment for Children, Frenchay Dysarthria Assessment, Schedule for Oral Motor Assessment) and swallowing (Paramatta Hospital's Assessment for Dysphagia) assessments, an outcome measure for swallowing (Royal Brisbane Hospital's Outcome Measure for Swallowing), and a cognitive rating scale (Rancho Level of Cognitive Functioning Scale). RESULTS: Across the patient group, oral-motor deficits resolved to normal status between 3 and 11 weeks post-referral (and at an average of 12 weeks post injury) and swallowing function and resolution to normal diet status were achieved by 3-11 weeks post-referral (and at an average of 12 weeks post-injury). The resolution of dysphagia and the resolution of oral-motor impairment and cognitive impairment were all highly correlated. CONCLUSION: The provision of a preliminary profile of oral-motor functioning and dysphagia resolution, and data on the linear relationship between swallowing impairment and cognition, will provide baseline information on the course of rehabilitation of dysphagia in the paediatric population post-TBI. Such data will contribute to more informed service provision and rehabilitation planning for paediatric patients post-TBI. PMID- 14742151 TI - Classifying psychiatric disorders after traumatic brain injury and orthopaedic injury in children: adequacy of K-SADS versus CBCL. AB - PRIMARY OBJECTIVE: [corrected] To evaluate the convergence between the Kiddie Schedule for Affective Disorders and Schizophrenia (K-SADS) for the diagnosis of psychiatric disorders and the Child Behavior Checklist (CBCL). RESEARCH DESIGN: Cross-sectional psychiatric study of 72 children with traumatic brain injuries or orthopaedic injuries aged 5-14. METHODS AND PROCEDURES: Sensitivity, specificity, total predictive value and odds ratio were calculated to evaluate the association between CBCL summary scores and K-SADS diagnosis of at least one psychiatric disorder (K-SADS-1), and of CBCL subscales and K-SADS diagnosis of Attention Deficit/Hyperactivity Disorder (ADHD). Lowered cut-off scores of t > or = 60 were used. MAIN OUTCOMES AND RESULTS: Convergence between the CBCL total problem scale and K-SADS-1 was poor. Convergence between 'at least one elevated CBCL subscale' and K-SADS-1, and between CBCL attention problems and social problems scales and K-SADS diagnosis of ADHD was excellent. CONCLUSIONS: Caution is needed when using the total problem scale of CBCL in predicting global psychological impairment because underestimation of problems is likely. The attention and social problems scales of CBCL can be used to estimate ADHD. PMID- 14742152 TI - Assessing stroke patients' prospective memory using virtual reality. AB - There is a dearth of empirical evidence about prospective memory (remembering to perform actions in the future) in stroke patients. A probable reason for this is that it is difficult to perform a realistic and controlled assessment of prospective memory ability in a rehabilitation setting. Virtual reality may provide a solution to this difficulty by allowing prospective memory to be tested in a simulation of a real-life situation whilst retaining a laboratory level of scientific control. This exploratory study assessed the performance of stroke patients and age-matched control participants on event-, time- and activity-based prospective memory retrieval tasks in a personal computer-based virtual environment. Stroke patients were severely impaired at the event- and activity based tasks compared with age-matched controls, but only marginally impaired at the time-based task. The additional knowledge gained from this form of assessment could direct rehabilitation more effectively towards specific impairments of individual patients. PMID- 14742153 TI - Novel approach to the application of botulinum toxin to the flexor digitorum superficialis muscle in acquired brain injury. AB - PRIMARY OBJECTIVE: To determine if the finger flexor mapping technique is useful in the reduction of spasticity when injecting neurotoxin within the flexor digitorum superficialis muscle. RESEARCH DESIGN: Case series of consecutive persons with acquired brain injury with upper limb spasticity. METHODS AND PROCEDURES: The flexor digitorum superficialis mapping technique was used to determine the optimal location for botulinum toxin A insertion into each belly of the flexor digitorum superficialis. Modified Ashworth Scale (MAS) ratings were recorded pre- and post-flexor digitorum superficialis injection from 17 patients (18 limbs) with upper limb spasticity. MAIN OUTCOMES AND RESULTS: The pre injection mean MAS score was 3.0 +/- 0.7 and the post-injection mean MAS score was 1.5 +/- 0.9 for this cohort. Using the Wilcoxon matched pairs signed rank test, the mean MAS finger flexor scores decreased significantly ( p < 0.05). No adverse events were noted with the procedure. CONCLUSIONS: This novel technique appears to be feasible and effective for placement of botulinum toxin in the treatment of finger flexor spasticity. Further studies are warranted to compare this method of administration with other injection approaches for the treatment of finger flexor spasticity. PMID- 14742161 TI - Prevalence of menstrual pain in relation to the reproductive life history of women from the Mayan rural community. AB - BACKGROUND: The prevalence of menstrual pain in Western societies is relatively high and has been shown to be related to many factors (e.g. age, menarche age, parity, menstrual cycle regularity, cigarette smoking, dietary habits). However, less data are available for traditional societies, within which there are, presumably, not as many potentially disturbing factors that could influence menstrual symptoms as in Western societies. AIM: The study seeks to determine the prevalence of dysmenorrhoea, and the factors related to menstrual pains in women with natural fertility (i.e. without hormonal forms of contraception). SUBJECTS AND METHODS: 177 non-smoking women between 18 and 45 years of age living in one Mayan village (Yaxcaba, Yucatan, Mexico) were the subjects of this study. The method of logistic regression was used with the dichotomous dependent variable being the presence or absence of menstrual pain together with such independent variables as age, age at menarche, number of pregnancies, age when giving birth to the first child, the weight at interview and BMI. RESULTS: The general prevalence of the menstrual pains was 28%. There was only one variable influencing dysmenorrhoea, namely the age at which women give birth to their first children. Those mothers who gave birth to their first child at an earlier age (at mean age of 19.4 vs 21.1 years) had a lower prevalence of dysmenorrhoea. CONCLUSION: In the studied traditional society the main factor related to menstrual pain was the age at which woman first gave birth. It is hypothesized that an earlier start to reproductive life in some way decreases the sensitivity of the uterus to prostaglandins. PMID- 14742162 TI - Mitochondrial DNA diversity in the Llanos de Moxos: Moxo, Movima and Yuracare Amerindian populations from Bolivia lowlands. AB - BACKGROUND: Movima, Yuracare, Ignaciano and Trinitario are Amerindian populations living in the Bolivian lowlands of the Amazonian basin. The cultural and genetic affinity of the peoples living in this area is poorly known, despite many archaeological studies demonstrating its importance in pre-Columbian times. Densely populated Amerindian groups occupied the region, both in the Llanos and along the river streams of the Amazonian basin, practising intense agricultural activities and exchange of goods. The historical and linguistic records indicate that the land was occupied through successive migrations that gave rise to complex socio-economic communities. Genetic information suggests that the colonization of the American continent was fairly simple from a emigrational point of view, but other evolutionary processes, such as genetic drift or natural selection, could have also shaped the genetic background of present day populations in the Beni region. AIM: The objective of this study is to characterize the genetic diversity of these populations by analysing the sequence variability of the HVR-I control region in the mitochondrial DNA (mtDNA). The Amerindian origin of these populations suggests that close genetic similarities should be evident between the Beni samples studied here and other Amerindian groups. However, complex processes of population interactions and/or isolation in the Beni region might result in non-expected genetic affinities. SUBJECTS AND METHODS: DNA was extracted from pulled-out hairs obtained in situ from non closely related individuals living in the Beni Department in Bolivia. DNA was extracted using a standard Chelex 100 method and a 401 bp DNA fragment of the HVR I region was amplified using specific primers (L-15978 and H-16412). DNA amplicons were purified by centrifugation using Microspin S-300 HR columns and both SNA strands were sequenced after asymmetric PCR using direct Dye-Terminator 2 sequencing kit (Perkin-Elmer). Two independent 401 and 328 bp DNA fragments were sequenced separately for each sample. The sequence analyses includes mismatch distributions and mean pairwise differences, median network analysis, and neighbour joining, maximum likelihood phylogenetic comparisons. Genetic diversity of DNA sequences was also measured in various ways for the sample studied and UPGMA trees were drawn, including a large number of South Amerindian sequences. RESULTS: The genetic diversity of 401 nucleotide long mtDNA sequences in the hypervariable control region, from positions 16,000-16,400, was characterized in a sample of 54 Amerindians living in the Llanos de Moxos. A total of 34 distinct lineages were observed, defined by 41 variable nucleotide positions, and 70.6% of all lineages were single sequences. All four major Amerindian haplogroups were detected (A 18.5%, n=10; B 24.1%, n=13, C 50.0% n=27; and D 5.6%, n=3). The median network analysis observed suggests that processes of population expansion took place in the Beni region. However, no clear haplotype differentiation by population could be detected. High levels of molecular variability and a bimodal pair-wise mismatch distribution were seen within the sample. The analyses of molecular variance (AMOVA) showed that most of the variance observed was due to intrapopulation variability, and that the highest among-groups variance was obtained when a linguistic classification criteria was used. The phylogenetic comparison revealed unique lineages in the Beni areas, not reported for other Amerindian populations. CONCLUSIONS: The genetic diversity observed in the Beni area is higher than that observed in other American populations living in much larger areas and with a long, known evolutionary history, despite the reduced area of Moxos. This could result from processes of reproductive isolation between groups, followed by population expansions and migration, where genetic drift might have be a major evolutionary force in population differentiation. PMID- 14742163 TI - Anthropometric indices in rural Malawians aged 45-75 years. AB - OBJECTIVE: The aim of this cross-sectional study was to assess anthropometric parameters of rural people aged 45-75 years in the Southern Region of Malawi. SUBJECTS: 97 males and 101 females aged 45-75 years were selected from 10 villages in Balaka district using a stratified cluster sampling method. Most responders were small-scale farmers growing maize and vegetables for household consumption. The anthropometric measurements taken were height, weight, triceps skinfold (TSK) and mid-upper arm circumference (MUAC). The derivatives from anthropometric measurements included body mass index (BMI), mid-upper arm muscle area (MUAMA) and mid-upper arm fat area (MUAFA). Weight status of Malawian subjects was estimated using WHO (1995) categories for BMI values, and height, weight, TSK, MUAC, MUAMA and MUAFA were compared with NHANES reference data for black sample. RESULTS: Correlation between age of subjects and the anthropometric indices was weak and insignificant. Males were generally taller and lighter than females. All anthropometric indices of Malawian subjects were considerably lower than NHANES medians. However the mean values of height, weight, MUAC and MUAMA of females were closer to the reference values than those of males. It was found that 9.1% of Malawian subjects had mild thinness (BMI 17.0-18.4 kg m(-2)), 13.6% had moderate thinness (BMI 16.0-16.9 kg m(-2)) and 18.2% exhibited severe thinness (BMI<16.0 kg m(-2)). The proportion of thin persons was higher (chi2 = 9.58, n = 1, p<0.01) among males than females (50.0% versus 35.7%). CONCLUSIONS: This study indicates that undernutrition is a serious problem among rural Malawians aged 45-75 years. PMID- 14742164 TI - Consanguinity, intracommunity and intercommunity marriages in a population sample of Israeli Jews. AB - BACKGROUND: Changes in the marriage patterns of Israeli Jews have been associated with the mass immigration of Jews from many countries over a relatively short period of time. AIM: This study seeks to document consanguineous, intracommunity and intercommunity marriage patterns, and to observe the changes that have occurred over time, and in relation to the level of education and religiousness. SUBJECT AND METHODS: During 1990-1992, 4388 Jewish women were interviewed after delivery in maternity wards throughout Israel. Demographic information was received, with special emphasis on country of origin, community and consanguinity of the couples and their parents. RESULTS: The consanguinity rate among the couples was 2.3%, including 0.8% first cousin marriages, with the highest consanguinity rate among Eastern Jews (7.1%). The rate of intracommunity marriages was 64% (25% Ashkenazim, 22% Sephardim and 17% Eastern Jews). The rate of intercommunity marriages was lowest among Ashkenazim. It rose with the level of education and inversely to the degree of religiousness. CONCLUSION: Over the past decades there has been a decline in consanguineous and intracommunity marriage rates and an increase in intercommunity marriages. Immigrant and ultraorthodox women tended to marry within the community as opposed to Israeli born women and those with higher educational level who tended to intermarry with other communities as well. PMID- 14742165 TI - Ambulatory blood pressure and heart rate in relation to hot flash experience among women of menopausal age. AB - OBJECTIVE: The purpose of this study was to evaluate whether ambulatory blood pressure and heart rate varied with hot flash experience among women of menopausal age. SUBJECTS AND METHODS: A total of 1149 ambulatory blood pressure and heart rate measurements from 20 women aged 44-55 were examined. Women were categorized by hot flash experience as (1) having had hot flashes during the study period (Symptomatic during Study; n = 5; 302 measurements), (2) having a past history of hot flashes, but no hot flashes during the study period (Historically Symptomatic; n = 7; 385 measurements), and (3) never having had a hot flash (Asymptomatic; n = 8; 462 measurements). Using repeated measures, nested ANOVA models that also adjusted for posture, the variation in blood pressure and heart rate associated with hot flash experience over the whole day and by location of measurement (microenvironment) was evaluated. RESULTS: The results show that, overall, systolic pressure did not differ among the hot flash experience groups although the Symptomatic during Study group had higher pressures at work than the other two groups (p<0.01), and tended to have higher pressures during sleep (p<0.08). The sleep diastolic pressure of the Asymptomatic group was significantly lower than that of the women who had hot flashes on the study day (p<0.01), but women who had a past history of hot flashes had slightly lower diastolic pressure (p<0.01) than those in the other two groups overall. Heart rates of the Asymptomatic group, however, were significantly lower (4-6 b.p.m.; p<0.001) in each microenvironment and over the whole day than both groups who had hot flash experience. CONCLUSIONS: These data suggest first that there may be a relationship between the experience of hot flashes and accelerated heart rate, and second that women who do not experience hot flashes may have lower sleep blood pressures than women who do. PMID- 14742166 TI - Pubertal development and sedentary behaviour during adolescence. AB - BACKGROUND: Assessing time spent in sedentary behaviours in relation to pubertal status, anthropometric differences, and body image will improve insight into the prevalence and determinants of such behaviours during adolescence. AIM: The study aimed to investigate the effects of age, puberty, gender, body composition, and sleep on sedentary behaviour. PARTICIPANTS AND METHODS: Participants were 64 boys and 55 girls in Year 6 (10.0-10.9 years of age), Year 8 (12.0-12.9 years) and Year 10 (14.0-14.9 years). Body mass index and percentage body fat were calculated from anthropometric measures. Sedentary behaviour and sleep time were computed using momentary-time sampling. Body image was measured using the Children's Physical Self-Perception Profile. Pubertal status was assessed from self-report of secondary sexual characteristics. RESULTS: After controlling for sleep time, no differences in sedentary time were seen for puberty onset or increased pubertal development. Correlations between pubertal status, body composition, and body image were stronger in girls than in boys. Correlations between body image and sedentary behaviour were not strong enough to infer behavioural choice differences. CONCLUSIONS: Reduced sleep time rather than changing body image and body composition during adolescence is associated with greater sedentary behaviour. PMID- 14742167 TI - Extremely limited mitochondrial DNA variability among the Ache Natives of Paraguay. AB - BACKGROUND: The Ache Natives are an especially interesting group of people, due to their distinctive morphological aspect and the fact that only in the last three decades have they established more permanent contact with outside populations. The objectives of the present study were: (a) to verify their distinctiveness in relation to mitochondrial DNA (mtDNA) variability; (b) to ascertain whether the pattern observed was congruent with other genetic studies performed among them; and (c) to establish historical inferences that would explain the eventual similarities or differences. SUBJECTS AND METHODS: Sample collection was made at two localities in eastern Paraguay. DNA from 64 maternally unrelated subjects were tested in relation to the mtDNA hypervariable segment 1 (HVS-1) by automatic sequencing. RESULTS: Fifty-six individuals presented exactly the same haplogroup B founder haplotype; another differed from it by a single transition polymorphism at site 16362, while six other subjects showed an identical haplogroup A founding haplotype. An A/G heteroplasmy at the 16269 site was seen in one haplogroup B individual, probably due to a somatic mutation. CONCLUSIONS: The Ache present distinctive differences and reduced mtDNA HVS-1 variability compared to other South American Natives. Similar differences were observed for other genetic systems. At present it is not clear whether their peculiarities already existed in their founding populations or whether they were secondarily acquired due to a long period of isolation in the humid, subtropical forest. PMID- 14742168 TI - Genetic variation at three VNTR loci (D1S80, APOB, and D17S5) in two tribal populations of Andhra Pradesh, India. AB - This study reports the genetic variation at three variable number of tandem repeat (VNTR) loci (APOB, D17S5 and D1S80) in two tribes (Thoti and Kolam) of Andhra Pradesh, India. Kolams constitute 1% of the total scheduled tribal population of Andhra Pradesh, while Thoti is a numerically small tribe. All three genetic loci were genotyped using the polymerase chain reaction (PCR) technique and were polymorphic in both populations. At the D1S80 locus, both populations showed higher frequencies of allele *31 (9-14%) than other Indian populations. In the APOB system, Thoti showed a very high frequency of allele *37 (54%) and for D17S5 system allele *4 was the most common in Thoti (32%) and allele *2 in Kolam (28%). Both tribes differed statistically significantly from other tribal populations of the region. The level of gene differentiation was low (GST = 0.038) for Indian tribal populations. The allele frequency distribution, heterozygosity and genetic diversity analysis shows that the observed genetic variation is socially and geographically structured. PMID- 14742169 TI - Y-STR polymorphisms among five Chinese minorities, Mosuo, Mongolian, Naxi, Pumi and Tibetan, in Yunnan Province, PR China. AB - Two hundred and thirty samples from five minority populations resident in Yunnan Province, PR China (the Mosuo, Mongolian, Naxi, Pumi and Tibetan communities) were analysed at eight polymorphic Y chromosome short tandem repeats (Y-STRs) loci. A total of 59 alleles were identified, with the largest number (n = 44) in the matriarchal Mosuo community. Two or three Y alleles were specific to each population. One hundred and thirty-five haplotypes were constructed, with 45 haplotypes in Tibetans, 34 in the Mosuo, 29 in Mongolians, 26 in the Pumi and 17 in the Naxi. The Tibetan and Mosuo peoples were closest in the neighbour-joining tree, whereas the Mongolians differed from the other four populations, reflecting their origins and present-day geographical location in Yunnan. PMID- 14742174 TI - Combined expectancy effects are modulated by the relation between expectancy cues. AB - Studies of combined expectancies have shown that spatial cueing effects are reduced on trials on which participants have to respond with an unexpected motor response. In the first two experiments the range of reduced expectancy effects is examined. Advance knowledge of the likely response was combined in a trial-by trial procedure with modality cueing, object cueing, and task cueing. Effects of modality cueing were reduced on trials on which the target requested an unexpected response. However, effects of object cueing as well as effects of task cueing were unaffected by response cueing. Comparing experiments revealed that different types of cues were used in different experiments. To test the effect of type of cue on the interaction of expectancies the third experiment combined spatial cueing with response cueing. When integrated cues were used that cued the likely target location by an arrow and the likely response by an arrow too, spatial cueing effects were reduced on trials with unexpected responses. However, spatial cueing effects remained unaffected by response cueing when separated cues were used consisting in a word cueing the response and an arrow cueing target location. An account for the modulation of combined expectancies by the relation between cues is suggested in terms of the adjusted expectancy model. PMID- 14742175 TI - ERP correlates of a receptive language-switching task. AB - Previous research has shown large response time costs (in excess of 50 ms) when bilingual speakers switch predictably back and forth between naming items (a productive switching task) in their first (L1) and second languages (L2). A recent study using event-related potentials (ERPs) has shown that switching between languages is associated with activity over frontal (N2) and parietal (late positive complex) areas of cortex (Jackson, Swainson, Cunnington, & Jackson, 2001). Switching between naming in different languages requires a switch in both language representations and language-specific motor responses. The current study investigated a receptive (input) language-switching task with a common manual response. Number words were presented in L1 and L2, and participants were required to judge whether the words were odd or even (a parity judgement). Response costs were considerably reduced, and the frontal and parietal switch related activity reported in the productive switching task was absent. Receptive switching was associated with early switch-related activity over central sensors that were not language specific. These results are discussed in relation to the idea that there is no language-specific lexical selection mechanism. Instead the costs of receptive language switching may arise from outside the bilingual lexicon. PMID- 14742176 TI - Evidence for disproportionate dual-task costs in older adults for episodic but not semantic memory. AB - Previous research demonstrates that older adults are poor at dual tasking, but there is less agreement on whether their decrement is worse than that predicted from single-task performance. This study investigated whether task domain moderates dual-task costs in old age. In two experiments, young and older adults retrieved either previously learned associates (episodic retrieval) or overlearned category members (semantic retrieval) under single or working-memory load conditions, using cued recall (Experiment 1) and recognition (Experiment 2) procedures. In both experiments the proportional costs of dual tasking were age invariant for semantic retrieval but were particularly marked for episodic retrieval, although the size of the age effect was reduced in recognition compared to cued recall. The data suggest that age effects in dual tasking may be domain specific. PMID- 14742177 TI - The role of sight of the hand in the development of prehension in childhood. AB - In two studies, children between 5 and 10 years of age were asked to reach to grasp an object without sight of the hand during the movement. The oldest children and adults were faster when they could see the hand and increased maximum grip aperture when they could not see the hand. The 10-year-olds were less able to integrate grasp and lift than adults when they could see their hands. Children aged 5 and 6 showed no increase in movement time when they could not see the hand and did not adapt maximum grip aperture to lack of sight. These effects remained when children were encouraged to reach for and lift the target as quickly as possible. The results indicate that younger children did not give preference to vision in the control of prehension, while older children used visual feedback to improve efficiency. Dependence on sight of the hand for the control of prehension does not simply decrease with age, but it may be integrated into an anticipatory control strategy where it contributes to the efficiency of control. PMID- 14742178 TI - Stimulus-driven attentional capture: An empirical comparison of display-size and distance methods. AB - Four experiments examined attentional capture by colour as assessed by two different investigative methods. Subjects performed a visual search task for a vertical-target line embedded among tilted-distractor lines, presented inside 4, 8, or 12 coloured discs. Interestingly, when the colour singleton was task irrelevant, and data were analysed by means of the display-size method combined with the zero-slope criterion, no evidence for attentional capture by colour was found. However, when data were analysed by means of the distance method, which consists of monitoring the spatial relationship between the target and the singleton, results showed that the target was found faster and/or more accurately when it was inside the singleton than when it was in a nonsingleton location. This provided evidence for a stimulus-driven attentional capture. In addition, the application of signal detection methodology showed that attentional capture, as revealed by the distance method, resulted from a perceptual modulation at the singleton location, rather than from a criterion shift. We conclude that, at least with the kind of stimuli used here, the display-size method combined with the zero-slope criterion is less than ideal for investigating how static discontinuities can affect the automatic deployment of visual attention. PMID- 14742179 TI - Beyond the two-strategy model of skilled spelling: effects of consistency, grain size, and orthographic redundancy. AB - Most models of spelling assume that people rely on two procedures when engaging in spelling: a lexical look-up procedure that retrieves spellings in their entirety, and a nonlexical procedure that constructs spellings with a set of phoneme-grapheme rules. In the present research, we investigated whether larger sized subsyllabic relationships also play a role in spelling, and how they compare to small-sized phoneme-grapheme relationships. In addition, we investigated whether purely orthographic units can explain some of the variance typically attributed to the mapping between sound and spelling. To do this, we ran five spelling experiments, two using real words and three using nonwords. Results from the experiments showed that there were independent contributions of both phoneme-grapheme and larger sized subsyllabic sound-spelling relationships, although the effect of phoneme-grapheme-sized relationships was always stronger and more reliable than larger sized subsyllabic sound-spelling relationships. Purely orthographic effects were also shown to affect word spelling, but no significant effects were found with nonword spelling. Together, the results support the hypothesis that a major constraint on spelling comes from phoneme grapheme-sized relationships. PMID- 14742180 TI - The attentional blink is susceptible to concurrent perceptual processing demands. AB - In a rapid serial visual presentation stream processing of a first target (T1) impairs detection or identification of a second target (T2) that appears within 500 ms after T1. This effect characterizes the so-called attentional blink (AB). To evaluate contemporary information-processing accounts of the AB phenomenon in terms of the underlying processing mechanisms the present study examined the potential influence of Task 1 difficulty on the AB effect. To this end, T1 contrast and T1 response requirements were systematically varied across four experiments. Experiment 1 ruled out a mere sensory basis of the contrast manipulation on T2 performance. When only T2 had to be reported (Experiment 2) an AB effect occurred that was slightly modulated by T1 contrast. When report of both T1 and T2 was required in a standard AB task (Experiment 3), the magnitude of the AB depended to a larger extent on stimulus contrast, and it increased further when speeded T1 choice responses were additionally required (Experiment 4). On the basis of the present impact of Task 1 difficulty on the AB effect we conclude that processing limitations cause the AB phenomenon. We discuss such limitations in terms of perceptual (T1 consolidation) and central (response selection) bottleneck processes. PMID- 14742183 TI - Pharmacodynamics of S-3578, a novel cephem, in murine lung and systemic infection models. AB - S-3578 is a novel beta-lactam with enhanced activity against drug-resistant gram positive cocci such as methicillin-resistant Staphylococcus aureus (MRSA). We used murine penicillin-resistant Streptococcus pneumoniae lung infection and neutropenic murine systemic MRSA infection models to determine the pharmacokinetic (PK)-pharmacodynamic (PD) parameter that best correlated with efficacy. Pharmacokinetic studies revealed that the maximum concentration in serum/dose values for S-3578 and cefepime in plasma in the lung infection model were 1.21 to 1.54 and 0.97 to 1.29, respectively; those for S-3578 in plasma in the systemic infection model were 0.78 to 1.02. The area under the concentration time curve (AUC)/dose values for S-3578 and cefepime in plasma in the lung infection model were 0.98 to 1.13 and 0.77 to 1.04, respectively, and those for S 3578 in plasma in the systemic infection model were 1.03 to 1.11. The half-lives of S-3578 and cefepime in plasma in the lung infection model were 0.29 to 0.38 and 0.29 to 0.34, respectively, and those of S-3578 in plasma in the systemic infection model were 0.40 to 0.61. The time above the MIC was the PK-PD parameter that best correlated with efficacy in the murine lung infection model (R(2) = 84 and 92% for S-3578 and cefepime in plasma, respectively). There was a twofold increase in the dose of S-3578 in the systemic infection model compared to that in the pneumonia model, yet the AUCs were the same. This may be due to the different MICs for the two pathogens. PMID- 14742182 TI - Issues in pharmacokinetics and pharmacodynamics of anti-infective agents: kill curves versus MIC. PMID- 14742184 TI - In vitro and in vivo efficacies of the new triazole albaconazole against Cryptococcus neoformans. AB - The activity of albaconazole (UR-9825; J. Uriach & Cia. S.A., Barcelona, Spain) was compared to that of fluconazole against 12 isolates of Cryptococcus neoformans in vitro and against 1 isolate in vivo in a rabbit model of cryptococcal meningitis. Albaconazole was 100-fold more potent in vitro than fluconazole on a per-weight basis and was fungicidal at potentially relevant concentrations for two isolates. MICs ranged from 100,000 copies/ml; CD4 lymphocytes < 200 cells/mm(3)), PRO 542 mediated an 80% response rate and statistically significant approximately 0.5 log(10) mean reductions in viral load for 4 to 6 weeks posttreatment. Similar findings were obtained in an analysis of all (n = 11) advanced disease patients treated to date with single doses of PRO 542 ranging from 1 to 25 mg/kg. In addition, a significant correlation was observed between antiviral effects observed in vivo and viral susceptibility to PRO 542 in vitro. The findings support continued development of PRO 542 for salvage therapy of advanced HIV-1 disease. PMID- 14742191 TI - Pharmacokinetics of saquinavir plus low-dose ritonavir in human immunodeficiency virus-infected pregnant women. AB - The physiologic changes that occur during pregnancy make it difficult to predict antiretroviral pharmacokinetics (PKs), but few data exist on the PKs of protease inhibitors in human immunodeficiency virus (HIV)-infected pregnant women. The objective of the present study was to determine the PKs of ritonavir (RTV) enhanced saquinavir (SQV) in HIV-infected pregnant women by an area under the curve (AUC)-targeted approach. A phase I, formal PK evaluation was conducted with HIV-infected pregnant woman during gestation, during labor and delivery, and at 6 weeks postpartum. The SQV-RTV regimen was 800/100 mg twice a day (b.i.d.), and nucleoside analogs were administered concomitantly. The SQV exposure targeted was an AUC at 24 h of 10,000 ng. h/ml. Participants were evaluated for 12-h steady state PKs at each time period. Thirteen subjects completed the PK evaluations during gestation, 7 completed the PK evaluations at labor and delivery, and 12 completed the PK evaluations postpartum. The mean baseline weight was 67.4 kg, and the median length of gestation was 23.3 weeks. All subjects achieved SQV exposures in excess of the target AUC. The SQV AUCs at 12 h (AUC(12)s) during gestation (29,373 +/- 17,524 ng. h/ml [mean +/- standard deviation]), during labor and delivery (26,189 +/- 22,138 ng. h/ml), and during the postpartum period (35,376 +/- 26,379 ng. h/ml) were not significantly different. The mean values of the PK parameters for RTV were lower during gestation than during the postpartum period: for AUC(12), 7,811 and 13,127 ng. h/ml, respectively; for trough concentrations, 376 and 632 ng/ml, respectively; and for maximum concentrations, 1,256 and 2,252 ng/ml, respectively (P K(237) within the non-penicillin binding domain and V-->E(470) near the SDN(464) conserved penicillin-binding motif in the penicillin-binding domain in COL52, were important for high-level resistance. The highest level of resistance was observed when all four mutations were present. The emergence of PBP 2a-mediated resistance to beta-lactams that bind to PBP 2a with a high affinity is likely to require multiple mutations in mecA; chromosomal mutations appear to have a minor role. PMID- 14742195 TI - In vitro selection and characterization of resistance to macrolides and related antibiotics in Mycoplasma pneumoniae. AB - Macrolide-resistant mutants of Mycoplasma pneumoniae were selected in vitro from the susceptible reference strain M129, by 23 to 50 serial passages in subinhibitory concentrations of macrolides and related antibiotics, erythromycin A, azithromycin, josamycin, clindamycin, quinupristin, quinupristin-dalfopristin, pristinamycin, and telithromycin. Mutants for which the MICs are increased could be selected with all antibiotics except the streptogramin B quinupristin. Portions of genes encoding 23S rRNA (domains II and V) and ribosomal proteins L4 and L22 of mutants were amplified by PCR, and their nucleotide sequences were compared to those of the susceptible strain M129. No mutation could be detected in domain II of 23S rRNA. Two point mutations in domain V of 23S rRNA, C2611A and A2062G, were selected in the presence of erythromycin A, azithromycin, josamycin, quinupristin-dalfopristin, and telithromycin. Mutants selected in the presence of clindamycin and telithromycin harbored a single amino acid change (H70R or H70L, respectively) in ribosomal protein L4, whereas insertions of one, two, or three adjacent glycines at position 60 (M. pneumoniae numbering) were selected in the presence of both streptogramin combinations. Telithromycin was the sole antibiotic that selected for substitutions (P112R and A114T) and deletions ((111)IPRA(114)) in ribosomal protein L22. Three sequential mutational events in 23S rRNA and in both ribosomal proteins were required to categorize the strain as resistant to the ketolide. Azithromycin and erythromycin A were the only selector antibiotics that remained active (MICs, 0.06 and 1 micro g/ml, respectively) on their mutants selected after 50 passages. PMID- 14742196 TI - Genotyping of Plasmodium falciparum pyrimethamine resistance by matrix-assisted laser desorption-ionization time-of-flight mass spectrometry. AB - Increasing resistance, recrudescences, and treatment failure have led to the replacement of chloroquine with the combination of pyrimethamine (PYR) and sulfadoxine (SDX) as the first-line antimalarial drugs for treatment of uncomplicated Plasmodium falciparum malaria in several areas where this disease is endemic. The development of resistance to PYR-SDX is favored by incomplete treatment courses or by subtherapeutic levels in plasma. PYR-SDX resistance has been associated with several single-nucleotide polymorphisms (SNPs) in the P. falciparum dihydrofolate reductase (pfdhfr) and the P. falciparum dihydropteroate synthetase (pfdhps) genes. We have established assays based on matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) that conveniently allow the identification of SNPs associated with PYR resistance. Variants occurring at codon positions 16, 51, 59, and 108 of the pfdhfr gene were analyzed by MALDI-TOF MS in synthetic oligonucleotides to determine the detection threshold. In addition, 63 blood samples from subjects with P. falciparum parasitemia of various degrees were analyzed. The results were compared to those obtained by DNA sequencing of the respective gene fragment. The results of MALDI TOF MS and DNA sequencing were consistent in 40 samples. In 23 samples two or three pfdhfr variants were detected by MALDI-TOF assays, whereas DNA-sequencing revealed one variant only. Simultaneous detection of two different mutations by biplex assays was, in principle, feasible. As demonstrated by the example of PYR resistance, MALDI-TOF MS allows for rapid and automated high-throughput assessment of drug sensitivity in P. falciparum malaria. PMID- 14742197 TI - Reemergence of macrolide resistance in pharyngeal isolates of group a streptococci in southwestern Pennsylvania. AB - We previously reported on the emergence of macrolide-resistant pharyngeal isolates of group A streptococci (GAS) in our community. The purpose of the present study was to track longitudinal trends in macrolide resistance in these isolates in southwestern Pennsylvania. Testing for susceptibility to erythromycin and clindamycin was performed for all pharyngeal GAS isolates recovered at the Children's Hospital of Pittsburgh and a local pediatric practice between September 2001 and May 2002. Macrolide resistance phenotypes and genotypes were determined by double-disk diffusion and PCR, respectively. Strain relatedness was determined by field inversion gel electrophoresis and emm gene sequence typing. A total of 708 isolates of GAS were recovered during the study period; 68 (9.6%) were macrolide resistant, while all isolates were sensitive to clindamycin. The monthly prevalence of macrolide resistance ranged from 0 to 41%. Only 21 of 573 (3.7%) strains recovered from September 2001 through March 2002 were macrolide resistant. A sudden increase in the rate of macrolide resistance (47 of 135 isolates [35%]) was seen in April and May 2002. Sixty-two isolates demonstrated the M phenotype (resistance to macrolide antibiotics), and six isolates demonstrated the MLS(B) phenotype (resistance to most macrolide, lincosamide, and streptogramin B antibiotics); these isolates were confirmed to be mef(A) and erm(A), respectively. Three unique mef(A) clones and four unique erm(A) clones were identified among the resistant isolates. The MIC at which 50% of isolates are inhibited (MIC(50)) for the mef(A) strains was 16 micro g/ml, while the MIC(50) for erm(A) strains was 8 micro g/ml. The finding of high levels of macrolide resistance among pharyngeal isolates of GAS for a second successive year in our community raises the concern that this problem may be more common in the United States than was previously appreciated. Longitudinal surveillance of isolates from multiple centers is needed to define the prevalence of antimicrobial agent-resistant GAS in the United States. PMID- 14742198 TI - Mechanism of action of NB2001 and NB2030, novel antibacterial agents activated by beta-lactamases. AB - Two potent antibacterial agents designed to undergo enzyme-catalyzed therapeutic activation were evaluated for their mechanisms of action. The compounds, NB2001 and NB2030, contain a cephalosporin with a thienyl (NB2001) or a tetrazole (NB2030) ring at the C-7 position and are linked to the antibacterial triclosan at the C-3 position. The compounds exploit beta-lactamases to release triclosan through hydrolysis of the beta-lactam ring. Like cephalothin, NB2001 and NB2030 were hydrolyzed by class A beta-lactamases (Escherichia coli TEM-1 and, to a lesser degree, Staphylococcus aureus PC1) and class C beta-lactamases (Enterobacter cloacae P99 and E. coli AmpC) with comparable catalytic efficiencies (k(cat)/K(m)). They also bound to the penicillin-binding proteins of S. aureus and E. coli, but with reduced affinities relative to that of cephalothin. Accordingly, they produced a cell morphology in E. coli consistent with the toxophore rather than the beta-lactam being responsible for antibacterial activity. In biochemical assays, they inhibited the triclosan target enoyl reductase (FabI), with 50% inhibitory concentrations being markedly reduced relative to that of free triclosan. The transport of NB2001, NB2030, and triclosan was rapid, with significant accumulation of triclosan in both S. aureus and E. coli. Taken together, the results suggest that NB2001 and NB2030 act primarily as triclosan prodrugs in S. aureus and E. coli. PMID- 14742199 TI - The ybxI gene of Bacillus subtilis 168 encodes a class D beta-lactamase of low activity. AB - The ybxI gene of Bacillus subtilis 168 encodes a preprotein of 267 amino acid residues, including a putative signal peptide of 23 residues. The YbxI primary structure exhibits high similarity scores with two members of the superfamily of the serine penicillin-recognizing enzymes: the class D beta-lactamases and the hydrophilic carboxy-terminal domains of the BlaR and MecR penicillin receptors. To determine the function and the activity of this putative penicillin recognizing enzyme, we have subcloned the ybxI gene in the pET-26b expression vector. Transformation of Escherichia coli BL21(DE3) by the recombinant plasmid pCIP51 resulted in the export of the mature YbxI in the periplasm as a water soluble protein. The recombinant protein was purified to 95% homogeneity. YbxI interacts with several beta-lactam antibiotics and can hydrolyze some of them. YbxI is not inactivated by clavulanic acid. The YbxI function and its enzymatic activity in B. subtilis remain unknown. The acyl-enzyme obtained after incubation of YbxI with a fluorescent derivative of ampicillin can be detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, confirming that YbxI can be acylated by beta-lactam antibiotics. YbxI does not hydrolyze some of the standard substrates of D-alanyl-D-alanine peptidases, the targets of penicillin. YbxI belongs to the penicillin-recognizing enzyme family but has an activity intermediate between those of a penicillin-binding protein and a beta-lactamase. PMID- 14742200 TI - Plasmid-mediated 16S rRNA methylase in Serratia marcescens conferring high-level resistance to aminoglycosides. AB - Serratia marcescens S-95, which displayed an unusually high degree of resistance to aminoglycosides, including kanamycins and gentamicins, was isolated in 2002 from a patient in Japan. The resistance was mediated by a large plasmid which was nonconjugative but transferable to an Escherichia coli recipient by transformation. The gene responsible for the aminoglycoside resistance was cloned and sequenced. The deduced amino acid sequence of the resistance gene shared 82% identity with RmtA, which was recently identified as 16S rRNA methylase conferring high-level aminoglycoside resistance in Pseudomonas aeruginosa. Histidine-tagged recombinant protein showed methylation activity against E. coli 16S rRNA. The novel aminoglycoside resistance gene was therefore designated rmtB. The genetic environment of rmtB was further investigated. The sequence immediately upstream of rmtB contained the right end of transposon Tn3, including bla(TEM), while an open reading frame possibly encoding a transposase was identified downstream of the gene. This is the first report describing 16S rRNA methylase production in S. marcescens. The aminoglycoside resistance mechanism mediated by production of 16S rRNA methylase and subsequent ribosomal protection used to be confined to aminoglycoside-producing actinomycetes. However, it is now identified among pathogenic bacteria, including Enterobacteriaceae and P. aeruginosa in Japan. This is a cause for concern since other treatment options are often limited in patients requiring highly potent aminoglycosides such as amikacin and tobramycin. PMID- 14742201 TI - Effects of interferon, ribavirin, and iminosugar derivatives on cells persistently infected with noncytopathic bovine viral diarrhea virus. AB - Persistent infection with hepatitis C virus (HCV) is a major cause of chronic hepatitis in humans. In chronic carriers, the viral infection induces liver damage that predisposes the patient for cirrhosis and can lead to hepatocellular carcinoma. Current chemotherapies are limited to alpha interferon (IFN-alpha) used either alone or in combination with ribavirin (RBV). In addition to its limited efficacy, this treatment is frequently poorly tolerated because of its side effects. The urgently needed development of new drugs is made difficult by the lack of an in vitro or in vivo infectivity model, and no cell line has been found so far to reliably and reproducibly support HCV infection. For this reason, the closely related pestivirus bovine viral diarrhea virus (BVDV) has sometimes been used as a surrogate in vitro infectivity model. In this study we used an MDBK cell line persistently infected with noncytopathic BVDV to assess the antiviral effect of IFN-alpha and RBV, the two drugs currently in clinical use against HCV. The same system was then used to evaluate the potential of two classes of iminosugar derivates to clear noncytopathic BVDV infection from MDBK cells. We show that treatment with long-alkyl-chain deoxynojirimycin derivatives, which are inhibitors of the endoplasmic reticulum (ER)-resident alpha glucosidases, can greatly reduce the amount of secreted enveloped viral RNA. Long alkyl-chain deoxygalactonojirimycin derivatives, which do not inhibit ER alpha glucosidases, were less potent but still more effective in this system than IFN alpha or ribavirin. PMID- 14742202 TI - Genetic analysis and complete primary structure of microcin L. AB - Escherichia coli LR05, in addition to producing MccB17, J25, and D93, secretes microcin L, a newly discovered microcin that exhibits strong antibacterial activity against related Enterobacteriaceae, including Salmonella enterica serovars Typhimurium and Enteritidis. Microcin L was purified using a two-step procedure including solid-phase extraction and reverse-phase C(18) high performance liquid chromatography. A 4,901-bp region of the DNA plasmid of E. coli LR05 was sequenced revealing that the microcin L cluster consists of four genes, mclC, mclI, mclA, and mclB. The structural gene mclC encoded a 105-amino acid precursor with a 15-amino-acid N-terminal extension ending with a Gly-Ala motif upstream of the cleavage site. This motif is typical of the class II microcins and other gram-positive bacteriocins exported by ABC transporters. The mclI immunity gene was identified upstream of the mclC gene and encodes a 51 amino-acid protein with two potential transmembrane domains. Located on the reverse strand, two genes, mclA and mclB, encoded the proteins MclA and MclB, respectively. They bear strong relatedness with the ABC transporter proteins and accessory factors involved in the secretion of microcins H47, V, E492, and 24. The microcin L genetic system resembles the genetic organization of MccV. Furthermore the MccL primary structure has been determined. It is a 90-amino-acid peptide of 8,884 Da with two disulfide bridges. The N-terminal region has significant homologies with several gram-positive bacteriocins. The C-terminal 32 amino-acid sequence is 87.5% identical to that of MccV. Together, these results strongly indicate that microcin L is a gram-negative class II microcin. PMID- 14742203 TI - Streptomyces clavuligerus has a second copy of the proclavaminate amidinohydrolase gene. AB - Past genetic studies have indicated that the genes encoding early enzymes of clavulanic acid biosynthesis may be duplicated in Streptomyces clavuligerus. We observed cross-hybridizing bands upon Southern analyses of proclavaminate amidinohydrolase (pah)-defective mutant strains of S. clavuligerus screened with a pah-specific probe. The DNA fragment responsible for this cross hybridization was cloned and sequenced and shown to encode a second copy of the pah gene. The new pah gene (pah1) was 1,056 bp in length, and its sequence was 72% identical to that of the original pah gene (pah2). Disruption mutants with defects in pah1 showed no significant effects on production of clavulanic acid or any of the clavam metabolites with stereochemistries opposite that of clavulanic acid (5S clavams) produced by S. clavuligerus when they were grown on starch asparagine or soy medium. However, double mutants with defects in both pah1 and pah2 were defective in the production of both clavulanic acid and all of the 5S clavam metabolites. PMID- 14742204 TI - Pharmacokinetics of total and unbound ertapenem in healthy elderly subjects. AB - Ertapenem is a new once-a-day parenteral carbapenem antimicrobial agent. The pharmacokinetics of unbound and total concentrations of ertapenem in plasma were investigated in elderly subjects and compared with historical data from young adults. In a single- and multiple-dose study, healthy elderly males and females (n = 14) 65 years old or older were given a 1-g intravenous (i.v.) dose once daily for 7 days. Plasma and urine samples collected for 24 h on days 1 and 7 following administration of the 1-g doses were analyzed by reversed-phase high performance liquid chromatography. Areas under the concentration-time curve from 0 h to infinity (AUC(0- infinity )) for elderly females and males were similar following administration of 1-g single i.v. doses, and thus, the genders were pooled in subsequent analyses. Concentrations in plasma and the half-life of ertapenem were generally higher and longer, respectively, in elderly subjects than in young adults. The mean AUC(0- infinity ) of total ertapenem in the elderly was 39% higher than that in young subjects following administration of a 1-g dose. The differences were slightly greater for the mean AUC(0- infinity ) of unbound ertapenem (71%). The unbound fraction of ertapenem in elderly subjects ( approximately 5 to 11%) was generally greater than that in young adults ( approximately 5 to 8%). As in young adults, ertapenem did not accumulate upon multiple dosing in the elderly. The pharmacokinetics of ertapenem in elderly subjects, while slightly different from those in young adults, do not require a dosage adjustment for elderly patients. PMID- 14742205 TI - New class of bacterial phenylalanyl-tRNA synthetase inhibitors with high potency and broad-spectrum activity. AB - Phenylalanyl (Phe)-tRNA synthetase (Phe-RS) is an essential enzyme which catalyzes the transfer of phenylalanine to the Phe-specific transfer RNA (tRNA(Phe)), a key step in protein biosynthesis. Phenyl-thiazolylurea sulfonamides were identified as a novel class of potent inhibitors of bacterial Phe-RS by high-throughput screening and chemical variation of the screening hit. The compounds inhibit Phe-RS of Escherichia coli, Haemophilus influenzae, Streptococcus pneumoniae, and Staphylococcus aureus, with 50% inhibitory concentrations in the nanomolar range. Enzyme kinetic measurements demonstrated that the compounds bind competitively with respect to the natural substrate Phe. All derivatives are highly selective for the bacterial Phe-RS versus the corresponding mammalian cytoplasmic and human mitochondrial enzymes. Phenyl thiazolylurea-sulfonamides displayed good in vitro activity against Staphylococcus, Streptococcus, Haemophilus, and Moraxella strains, reaching MICs below 1 micro g/ml. The antibacterial activity was partly antagonized by increasing concentrations of Phe in the culture broth in accordance with the competitive binding mode. Further evidence that inhibition of tRNA(Phe) charging is the antibacterial principle of this compound class was obtained by proteome analysis of Bacillus subtilis. Here, the phenyl-thiazolylurea-sulfonamides induced a protein pattern indicative of the stringent response. In addition, an E. coli strain carrying a relA mutation and defective in stringent response was more susceptible than its isogenic relA(+) parent strain. In vivo efficacy was investigated in a murine S. aureus sepsis model and a S. pneumoniae sepsis model in rats. Treatment with the phenyl-thiazolylurea-sulfonamides reduced the bacterial titer in various organs by up to 3 log units, supporting the potential value of Phe-RS as a target in antibacterial therapy. PMID- 14742206 TI - Epidemiology of conjugative plasmid-mediated AmpC beta-lactamases in the United States. AB - A sample of 752 resistant Klebsiella pneumoniae, Klebsiella oxytoca, and Escherichia coli strains from 70 sites in 25 U.S. states and the District of Columbia was examined for transmissibility of resistance to ceftazidime and the nature of the plasmid-mediated beta-lactamase involved. Fifty-nine percent of the K. pneumoniae, 24% of the K. oxytoca, and 44% of the E. coli isolates transferred resistance to ceftazidime. Plasmids encoding AmpC-type beta-lactamase were found in 8.5% of the K. pneumoniae samples, 6.9% of the K. oxytoca samples, and 4% of the E. coli samples, at 20 of the 70 sites and in 10 of the 25 states. ACT-1 beta lactamase was found at eight sites, four of which were near New York City, where the ACT-1 enzyme was first discovered; ACT-1 beta-lactamase was also found in Massachusetts, Pennsylvania, and Virginia. FOX-5 beta-lactamase was also found at eight sites, mainly in southeastern states but also in New York. Two E. coli strains produced CMY-2, and one K. pneumoniae strain produced DHA-1 beta lactamase. Pulsed-field gel electrophoresis and plasmid analysis suggested that AmpC-mediated resistance spread both by strain and plasmid dissemination. All AmpC beta-lactamase-containing isolates were resistant to cefoxitin, but so were 11% of strains containing transmissible SHV- and TEM-type extended-spectrum beta lactamases. A beta-lactamase inhibitor test was helpful in distinguishing the two types of resistance but was not definitive since 24% of clinical isolates producing AmpC beta-lactamase had a positive response to clavulanic acid. Coexistence of AmpC and extended-spectrum beta-lactamases was the main reason for these discrepancies. Plasmid-mediated AmpC-type enzymes are thus responsible for an appreciable fraction of resistance in clinical isolates of Klebsiella spp. and E. coli, are disseminated around the United States, and are not so easily distinguished from other enzymes that mediate resistance to oxyimino-beta lactams. PMID- 14742207 TI - Enhanced saquinavir exposure in human immunodeficiency virus type 1-infected patients with diarrhea and/or wasting syndrome. AB - The protease inhibitor saquinavir was administered to 100 human immunodeficiency virus type 1 (HIV-1)-infected patients as a single 600-mg oral dose (hard gelatin capsules) with a standard breakfast, including 200 ml of grapefruit juice, during an open-label trial to assess whether diarrhea and/or wasting syndrome has consequences on its pharmacokinetics. Three groups of patients were enrolled: group 1, asymptomatic patients (n = 30); group 2, AIDS symptomatic patients without body weight loss or diarrhea (n = 37); and group 3, AIDS symptomatic patients with severe body weight loss and/or diarrhea (n = 33). Clinical and biological data (covariates) were collected. A population approach was performed with three blood samples per patient to estimate the mean population pharmacokinetic parameters (clearance [CL]/oral bioavailability [F], V/F, k(a), and lag time) and the derived ones (k(el), C(max), T(max), and area under the curve [AUC]). The relationships between groups, exposure (i.e., estimated individual post hoc AUCs), and covariates were explored by using multiple linear regressions. A significant increase in median AUCs (165, 349, and 705 ng. h. ml( 1) for groups 1, 2, and 3, respectively [P < 0.0001]) was observed. The enhancement in saquinavir exposure could be due to the destruction of the transporters in enterocytes and/or to the enlargement of their tight junctions, allowing a paracellular crossing of saquinavir as the illness spreads. Because of grapefruit juice intake by every patient, no implication of CYP3A4 could be assessed. These results strongly suggest that, despite its low intrinsic oral bioavailability, saquinavir can be considered as a relevant treatment for HIV-1 infected patients with diarrhea and/or wasting syndrome. This must be evaluated in a long-term period. PMID- 14742208 TI - Subinhibitory concentrations of linezolid reduce Staphylococcus aureus virulence factor expression. AB - The influence of the antibiotic linezolid on the secretion of exotoxins by Staphylococcus aureus was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis combined with matrix-assisted laser desorption ionization-time of flight mass spectrometry and Western blot analysis. S. aureus suspensions were treated with grading subinhibitory concentrations of linezolid (12.5, 25, 50, and 90% of MIC) at different stages of bacterial growth (i.e., an optical density at 540 nm [OD(540)] of 0.05 or 0.8). When added to S. aureus cultures at an OD(540) of 0.05, linezolid reduced in a dose-dependent manner the secretion of specific virulence factors, including staphylococcal enterotoxin A (SEA) and SEB, bifunctional autolysin, autolysin, protein A, and alpha- and beta-hemolysins. In contrast, other presumably nontoxic exoproteins remained unchanged or even accumulated in supernatants in the presence of linezolid at a 90% MIC. Similarly, when added at OD(540) of 0.8, that is, after quorum sensing, linezolid reduced the release of virulence factors, whereas the relative abundance of nontoxic exoproteins such as triacylglycerol lipase, glycerol ester hydrolase, DnaK, or translation elongation factor EF-Tu was found to be increased. Consistently, linezolid reduced in a dose-dependent manner the tumor necrosis factor-inducing activity secreted by S. aureus into the culture supernatants. The results of our study suggest that the expression of virulence factors in S. aureus is especially sensitive to the inhibition of protein synthesis by linezolid, which should be an advantage in the treatment of infections with toxin-producing S. aureus. PMID- 14742209 TI - Duplex real-time PCR assay for rapid detection of ampicillin-resistant Enterococcus faecium. AB - Rapid and accurate identification of carriers of resistant microorganisms is an important aspect of efficient infection control in hospitals. Traditional identification methods of antibiotic-resistant bacteria usually take at least 3 to 4 days after sampling. A duplex real-time PCR assay was developed for rapid detection of ampicillin-resistant Enterococcus faecium (ARE). Primers and probes that are used in this assay specifically detected the D-Ala-D-Ala ligase gene of E. faecium and the modified penicillin-binding protein 5 gene (pbp5) carrying the Glu-to-Val substitution at position 629 (Val-629) in a set of 129 tested E. faecium strains with known pbp5 sequence. Presence of the Val-629 in the strain set from 11 different countries was highly correlated with ampicillin resistance. In a screening of hospitalized patients, the real-time PCR assay yielded a sensitivity and a specificity for the detection of ARE colonization of 95% and 100%, respectively. The results were obtained 4 h after samples were harvested from overnight broth of rectal swab samples, identifying both species and the resistance marker mutation in pbp5. This novel assay reliably identifies ARE 2 to 3 days more quickly than traditional culture methods, thereby increasing laboratory throughput, making it useful for rectal screening of ARE. The assay demonstrates the advantages of real-time PCR for detection of nosocomial pathogens. PMID- 14742211 TI - Three-dimensional models of wild-type and mutated forms of cytochrome P450 14alpha-sterol demethylases from Aspergillus fumigatus and Candida albicans provide insights into posaconazole binding. AB - The cytochrome P450 sterol 14alpha-demethylase enzyme (CYP51) is the target of azole antifungals. Azoles block ergosterol synthesis, and thereby fungal growth, by binding in the active-site cavity of the enzyme and ligating the iron atom of the heme cofactor through a nitrogen atom of the azole. Mutations in and around the CYP51 active site have resulted in azole resistance. In this work, homology models of the CYP51 enzymes from Aspergillus fumigatus and Candida albicans were constructed based on the X-ray crystal structure of CYP51 from Mycobacterium tuberculosis. Using these models, binding modes for voriconazole (VOR), fluconazole (FLZ), itraconazole (ITZ), and posaconazole (POS) were predicted from docking calculations. Previous work had demonstrated that mutations in the vicinity of the heme cofactor had a greater impact on the binding of FLZ and VOR than on the binding of POS and ITZ. Our modeling data suggest that the long side chains of POS and ITZ occupy a specific channel within CYP51 and that this additional interaction, which is not available to VOR and FLZ, serves to stabilize the binding of these azoles to the mutated CYP51 proteins. The model also predicts that mutations that were previously shown to specifically impact POS susceptibility in A. fumigatus and C. albicans act by interfering with the binding of the long side chain. PMID- 14742210 TI - Differential gene expression in auristatin PHE-treated Cryptococcus neoformans. AB - The antifungal pentapeptide auristatin PHE was recently shown to interfere with microtubule dynamics and nuclear and cellular division in the opportunistic pathogen Cryptococcus neoformans. To gain a broader understanding of the cellular response of C. neoformans to auristatin PHE, mRNA differential display (DD) and reverse transcriptase PCR (RT-PCR) were applied. Examination of approximately 60% of the cell transcriptome from cells treated with 1.5 times the MIC (7.89 micro M) of auristatin PHE for 90 min revealed 29 transcript expression differences between control and drug-treated populations. Differential expression of seven of the transcripts was confirmed by RT-PCR, as was drug-dependent modulation of an additional seven transcripts by RT-PCR only. Among genes found to be differentially expressed were those encoding proteins involved in transport, cell cycle regulation, signal transduction, cell stress, DNA repair, nucleotide metabolism, and capsule production. For example, RHO1 and an open reading frame (ORF) encoding a protein with 91% similarity to the Schizophyllum commune 14-3-3 protein, both involved in cell cycle regulation, were down-regulated, as was the gene encoding the multidrug efflux pump Afr1p. An ORF encoding a protein with 57% identity to the heat shock protein HSP104 in Pleurotus sajor-caju was up regulated. Also, three transcripts of unknown function were responsive to auristatin PHE, which may eventually contribute to the elucidation of the function of their gene products. Further study of these differentially expressed genes and expression of their corresponding proteins are warranted to evaluate how they may be involved in the mechanism of action of auristatin PHE. This information may also contribute to an explanation of the selectivity of auristatin PHE for C. neoformans. This is the first report of drug action using DD in C. neoformans. PMID- 14742212 TI - Functional angucycline-like antibiotic gene cluster in the terminal inverted repeats of the Streptomyces ambofaciens linear chromosome. AB - Streptomyces ambofaciens has an 8-Mb linear chromosome ending in 200-kb terminal inverted repeats. Analysis of the F6 cosmid overlapping the terminal inverted repeats revealed a locus similar to type II polyketide synthase (PKS) gene clusters. Sequence analysis identified 26 open reading frames, including genes encoding the beta-ketoacyl synthase (KS), chain length factor (CLF), and acyl carrier protein (ACP) that make up the minimal PKS. These KS, CLF, and ACP subunits are highly homologous to minimal PKS subunits involved in the biosynthesis of angucycline antibiotics. The genes encoding the KS and ACP subunits are transcribed constitutively but show a remarkable increase in expression after entering transition phase. Five genes, including those encoding the minimal PKS, were replaced by resistance markers to generate single and double mutants (replacement in one and both terminal inverted repeats). Double mutants were unable to produce either diffusible orange pigment or antibacterial activity against Bacillus subtilis. Single mutants showed an intermediate phenotype, suggesting that each copy of the cluster was functional. Transformation of double mutants with a conjugative and integrative form of F6 partially restored both phenotypes. The pigmented and antibacterial compounds were shown to be two distinct molecules produced from the same biosynthetic pathway. High-pressure liquid chromatography analysis of culture extracts from wild-type and double mutants revealed a peak with an associated bioactivity that was absent from the mutants. Two additional genes encoding KS and CLF were present in the cluster. However, disruption of the second KS gene had no effect on either pigment or antibiotic production. PMID- 14742213 TI - Determination of ddATP levels in human immunodeficiency virus-infected patients treated with dideoxyinosine. AB - Clinical failures of the highly active antiretroviral therapy could result from inefficient intracellular concentrations of antiviral drugs. The determination of drug contents in target cells of each patient would be useful in clinical investigations and trials. The purpose of this work was to quantify the intracellular concentration of ddATP, the active metabolite of dideoxyinosine (ddI), in peripheral blood mononuclear cells (PBMCs) of human immunodeficiency virus (HIV)-infected patients treated with ddI. We have raised antibodies against ddA-citrate, a stable isostere of ddATP selected on the basis of its structural and electronic analogies with ddATP. The anti-ddA-citrate antibodies recognized ddATP and ddA with nanomolar affinities and cross-reacted neither with any of the nucleotide reverse transcriptase inhibitors used in HIV therapy nor with their phosphorylated metabolites. The three phosphorylated metabolites of ddI (ddAMP, ddADP, and ddATP) were purified by anion exchange chromatography and the amount of each metabolite was determined by radioimmunoassay with or without prior phosphatase treatment. The intracellular levels of the three ddI metabolites were measured both in an in vitro model and in PBMCs of HIV-infected patients under ddI treatment. The possibility to measure intracellular levels of ddATP from small blood samples of HIV-infected patients treated with ddI could be exploited to develop individual therapeutic monitoring. PMID- 14742214 TI - Multiplex PCR amplimer conformation analysis for rapid detection of gyrA mutations in fluoroquinolone-resistant Mycobacterium tuberculosis clinical isolates. AB - A new strategy known as multiplex PCR amplimer conformation was developed for detection of mutation in the gyrA gene of 138 clinical isolates of Mycobacterium tuberculosis. The method generated a single-stranded and heteroduplex DNA banding pattern of multiplex PCR amplimers of the region of interest that was extremely sensitive to specific mutations, thus enabling much more sensitive and reliable mutation analysis compared to the standard single-stranded conformation polymorphism technique. The genetic profiles of the gyrA gene of the 138 isolates as detected by MPAC were confirmed by nucleotide sequencing and were found to correlate strongly with the in vitro susceptibilities of the mutant strains to six fluoroquinolones (ofloxacin, levofloxacin, sparfloxacin, moxifloxacin, gatifloxacin, and sitafloxacin). All 32 isolates that contained gyrA mutations exhibited cross-resistance to the six fluoroquinolones (ofloxacin MIC for 90% of strains > 16 mg/liter), although moxifloxacin, gatifloxacin, and sitafloxacin (MIC for 90% of strains /==" BORDER="0"> 16 mg/liter). All gyrA mutations were clustered in codons 90, 91, and 94, and aspartic acid 94 was most frequently mutated. Twenty-three isolates without gyrA mutations were also found to exhibit reduced susceptibility to ofloxacin (MIC for 90% of strains = 4 mg/liter), but largely remained susceptible to other drugs (MIC for 90% of strains 2 micro g/ml, which was higher than the MIC at which 90% of Staphylococcus epidermidis strains are inhibited. PMID- 14742236 TI - Antimicrobial activity and bacterial-membrane interaction of ovine-derived cathelicidins. AB - Three ovine-derived cathelicidins, SMAP29, OaBac5mini, and OaBac7.5mini, were compared with respect to their antibacterial activities and interactions with membranes. SMAP29 was confirmed to be alpha-helical, broad spectrum, and able to disrupt both the outer and the cytoplasmic membranes at relatively low concentrations. In contrast, the two proline- and arginine-rich OaBac peptides had more-modest antibacterial activities, reduced levels of lipopolysaccharide binding, and a lesser ability to depolarize the cytoplasmic membrane, consistent with a cytoplasmic target. PMID- 14742237 TI - Telithromycin susceptibility and genomic diversity of macrolide-resistant serotype III group B streptococci isolated in perinatal infections. AB - We studied the telithromycin, erythromycin, azithromycin, and clindamycin susceptibilities of serotype III macrolide-resistant group B streptococci, together with genetic mechanisms of resistance and genomic diversity. ermB, ermA, and mefA were found in, respectively, 57, 32, and 9% of isolates. The telithromycin MIC at which 90% of isolates were inhibited was 0.5 micro g/ml. Macrolide resistance was associated with dissemination of resistance determinants among isolates of different genetic backgrounds. PMID- 14742238 TI - Clonal multidrug-resistant Shigella dysenteriae type 1 strains associated with epidemic and sporadic dysenteries in eastern India. AB - Multidrug-resistant strains of Shigella dysenteriae type 1 were implicated in three outbreaks and sporadic cases of dysentery in eastern India in 2002 and 2003. After a hiatus of 14 years, this pathogen reemerged with an altered antibiotic resistance pattern. In addition to ampicillin, co-trimoxazole, tetracycline, chloramphenicol, and nalidixic acid, all the recent strains were resistant to norfloxacin, lomefloxacin, pefloxacin, and ofloxacin and showed reduced susceptibility to ciprofloxacin. Pulsed-field gel electrophoresis identified a new clone of S. dysenteriae type 1 that was associated with the recent outbreaks and sporadic cases. Based on the spatial and temporal spread of multidrug-resistant S. dysenteriae type 1, we predict that this clonal type may spread further in this region. PMID- 14742239 TI - Effects of positively selected sequence variations in human and Macaca fascicularis beta-defensins 2 on antimicrobial activity. AB - The evolution of orthologous genes coding for beta-defensin 2 (BD2) in primates has been subject to positive selection during the divergence of the platyrrhines from the catarrhines and of the Cercopithecidae from the Hylobatidae, great apes, and humans. Three peptides have been selected for a functional analysis of the effects of sequence variations on the direct antimicrobial activity: human BD2 (hBD2), Macaca fascicularis BD2 (mfaBD2), and a variant of the human peptide lacking Asp(4), (-D)hBD2, which is characteristic only of the human/great ape peptides. hBD2 and mfaBD2 showed a significant difference in specificity, the former being more active towards Escherichia coli and the later towards Staphylococcus aureus and Candida albicans. Asp(4) in the human peptide appears to be important, as (-D)hBD2 was less structured and had a markedly lower antimicrobial activity. The evolution of beta-defensin 2 in primates may thus have been driven, at least in part, by different environmental pressures so as to modulate antimicrobial activity. PMID- 14742240 TI - First Canadian Salmonella enterica serovar typhi isolate harboring an integron. PMID- 14742241 TI - Dysregulation of mitogen-activated protein kinase signaling pathways in simian immunodeficiency virus encephalitis. AB - Central nervous system (CNS) disease is a frequent complication of human immunodeficiency virus (HIV)-1 infection. Identification of cellular mechanisms that control virus replication and that mediate development of HIV-associated neuropathology will provide novel strategies for therapeutic intervention. The milieu of the CNS during HIV infection is extraordinarily complex because of infiltration of inflammatory cells and production of chemokines, cytokines, and neurotoxic molecules. Cells in the CNS must integrate signaling pathways activated simultaneously by products of virus replication and infiltrating immune cells. In this study, we examined activation of mitogen-activated protein kinases (MAPKs) in the CNS of simian immunodeficiency virus-infected macaques during acute, asymptomatic, and terminal infection. We demonstrate that significantly increased (P < 0.02) activation of ERK MAPK, typically associated with anti apoptotic and neuroprotective pathways, occurs predominantly in astrocytes and immediately precedes suppression of virus replication and macrophage activation that occur after acute infection. In contrast, significantly increased activation of proapoptotic, neurodegenerative MAPKs JNK (P = 0.03; predominantly in macrophages/microglia), and p38 (P = 0.03; predominantly in neurons and astrocytes) after acute infection correlates with subsequent resurgent virus replication and development of neurological lesions. This shift from classically neuroprotective to neurodegenerative MAPK pathways suggests that agents that inhibit activation of JNK/p38 may be protective against HIV-associated CNS disease. PMID- 14742242 TI - Bystander CD8 T-cell-mediated demyelination is interferon-gamma-dependent in a coronavirus model of multiple sclerosis. AB - Mice infected with the coronavirus mouse hepatitis virus, strain JHM (JHM) develop a disease that shares many histological characteristics with multiple sclerosis. We previously demonstrated that JHM-infected mice that only have CD8 T cells specific for an epitope not in the virus develop demyelination on specific activation of these cells. Herein we show that this process of bystander T-cell mediated demyelination is interferon-gamma (IFN-gamma)-dependent. The absence of IFN-gamma abrogated demyelination but did not change T-cell infiltration or expression levels of inflammatory cytokines or chemokines in the spinal cord. These results are consistent with models in which IFN-gamma contributes to CD8 T cell-mediated demyelination by activation of macrophages/microglia, the final effector cells in the disease process. PMID- 14742243 TI - Transgene expression of alpha(1,2)-fucosyltransferase-I (FUT1) in tumor cells selectively inhibits sialyl-Lewis x expression and binding to E-selectin without affecting synthesis of sialyl-Lewis a or binding to P-selectin. AB - During inflammation, E- and P-selectins appear on activated endothelial cells to interact with leukocytes through sialyl-Lewis x and sialyl-Lewis a antigens (sLe(x/a)). These selectins can also interact with tumor cells in a sialyl-Lewis dependent manner and for this reason, they are thought to play a key role in metastasis. Diverting the biosynthesis of sialyl-Lewis antigens toward nonadhesive structures is an attractive gene therapy for preventing the hematogenous metastatic spread of cancers. We have previously shown that transfection of alpha(1,2)-fucosyltransferase-I (FUT1) in Chinese hamster ovary (CHO) cells had a slight effect on the overall sialylation while the synthesis of sLE(x) was dramatically prevented. We herein delivered the gene of FUT1 by a human immunodeficiency virus-derived lentiviral vector to three human cancer cell lines including pancreatic (BxPC3), hepatic (HepG2), and colonic (HT-29) cancer cells. We found that on FUT1 transduction, all cells exhibited a dramatic decrease in sLe(x) synthesis with a concomitant increase in Le(y) and Le(b) expression, without any detectable effect on the level of cell surface sLe(a) antigens. In parallel, FUT1-transduced HT-29 and HepG2 cells, but not BxPC3 cells, failed to interact with E-selectin as assessed by E-selectin-binding assay or dynamic adhesion to activated endothelial cells. We show also that transduced FUT1 efficiently fucosylates the P-selectin ligand PSGL-1 without altering P selectin binding. These results have important implications for understanding cell-specific reactions underlying the synthesis of selectin ligands in cancer cells and may provide a basis for the development of anti-metastatic gene therapy. PMID- 14742244 TI - Leptin-deficient mice are protected from accelerated nephrotoxic nephritis. AB - Leptin is an adipose tissue-derived hormone that signals nutritional status to the hypothalamus. Recent evidence indicates that leptin modifies proinflammatory immune responses and may provide a key link between nutritional deficiency and immune dysfunction. To study the influence of leptin deficiency on immune mediated renal disease, susceptibility to accelerated nephrotoxic nephritis was examined in leptin-deficient C57BL/6-ob/ob mice and C57BL/6 wild-type controls. The model was induced with sheep anti-mouse glomerular basement membrane antibody injected to mice preimmunized against sheep IgG, and mice were sacrificed 8 days after induction of disease. The leptin-deficient ob/ob mice were strongly protected from glomerular crescent formation, macrophage infiltration, glomerular thrombosis, and albuminuria in this model. Our findings suggest that leptin is required for the induction and maintenance of immune-mediated glomerulonephritis, and that blockade of the leptin axis might provide an attractive therapeutic possibility in human autoimmune disease. PMID- 14742245 TI - Higher vessel densities in retinoblastoma with local invasive growth and metastasis. AB - In this study, the importance of angiogenesis (the growth of new blood vessels from existing ones) for the growth of retinoblastoma was investigated by a retrospective immunohistochemical analysis. An individual vessel index for each tumor was determined using the endothelial-specific antibody CD 31 for vessel staining. The obtained data were correlated with clinical features, pathohistological characteristics, and the presence of metastasis. In 107 retinoblastomas collected between 1980 and 1990, we found no difference in the vessel densities between uni- and bilateral retinoblastomas (P = 0.41). However, tumors that had invaded the chorioid and/or the optic nerve statistically showed higher vessel densities than tumors without local invasive growth (P = 0.05 and P = 0.024). A tendency of higher vessel densities in retinoblastomas presenting with metastasis at the time of diagnosis was observed (P = 0.11). Based on this observation, we proceeded to examine all retinoblastomas presenting with metastasis at the time of diagnosis. These included patients that were treated between 1968 and 1993. The 18 investigated retinoblastomas had significantly higher vessel densities than all other retinoblastomas presenting without metastasis (P = 0.025). Our data indicate that in retinoblastoma, blood vessels are essential for local and systemic invasive growth. Therefore, an anti angiogenic therapy could be considered in the multimodal therapy concept for retinoblastomas with invasive growth, both locally and systemically. PMID- 14742246 TI - Intermediate filaments control the intracellular distribution of caspases during apoptosis. AB - Caspases are responsible for a cascade of events controlling the disassembly of apoptotic cells. We now demonstrate that caspase-9 is activated at an early stage of apoptosis in epithelial cells and all its detectable, catalytically active large subunits (both the p35 and p37) are concentrated on cytokeratin fibrils. Immunolabeling of distinctive neoepitopes, exposed by cleavage of procaspase-9 at either Asp315 or Asp330, was co-localized on these fibrils with active caspase-3, caspase-cleaved cytokeratin-18, death-effector-domain containing DNA-binding protein and ubiquitin. Cytokeratin filaments may thus provide a scaffold whereby active subunits of caspase-9 can activate caspase-3 which, in turn, can activate more caspase-9 so forming an amplification loop to facilitate cleavage of cytokeratin-18, disruption of the cytoskeleton and the ensuing formation of cytoplasmic inclusions. These inclusions, formed from the collapse of fibrils, together with their associated components, also contain ubiquitinated proteins, vimentin, heat-shock protein 72, and tumor necrosis factor receptor type-1 associated death domain protein. Many of their constituents, including active caspases, remain sequestered within these inclusions, even after detergent treatment and isolation. Thus, such inclusions do not merely accumulate disrupted cytokeratins but also sequestrate potentially noxious proteins that could injure healthy neighboring cells. PMID- 14742247 TI - Plasma cell-like morphology of Th1-cytokine-producing cells associated with the loss of CD3 expression. AB - Here we clarified the morphology and phenotype of interleukin (IL)-17- and interferon (IFN)-gamma-producing cells in both in vitro and in vivo situations. Oligoclonal activation of normal peripheral blood mononuclear cells with the superantigen Staphylococcus aureus enterotoxin B and polyclonal activation with phorbol myristate acetate/phytohemagglutinin were used as in vitro models. This study was extended to various in vivo situations such as rheumatoid arthritis, dermatomyositis, and normal activated lymph nodes. The phenotype of IL-17- and IFN-gamma-producing cells was evaluated by immunohistochemistry using the CD3 and CD4 T-cell markers, the CD20, CD38, kappa and lambda light chain B-cell lineage markers. The expression of two chemokine receptors, CCR6 and CCR7, involved with their associated ligands CCL20 and CCL19/CCL21 in the migration of T lymphocytes, was evaluated in tissue sections. After both polyclonal and oligoclonal activation, IL-17+ and IFN-gamma+ cells acquired a plasma cell-like morphology associated with a high secretory activity, the reduced expression of CD3, and no change of CD4 expression. In rheumatoid arthritis, dermatomyositis, and activated lymph nodes, both IL-17- and IFN-gamma-producing cells had the same morphology. These Th1 cytokine-producing cells were CD4(+)-, CD3-, and B-cell lineage marker negative. In both in vitro and in vivo situations, expression of CCR6 or CCR7 was not associated with a particular subset. In conclusion, activated T-helper CD4(+) T cells, by their release of cytokines, seem to have functional similarities with plasma cells secreting immunoglobulins. PMID- 14742248 TI - Differential distribution of the JC virus receptor-type sialic acid in normal human tissues. AB - JC virus (JCV), a member of the polyomavirus family, causes a demyelinating disease of the central nervous system (CNS) in humans known as progressive multifocal leukoencephalopathy. Although glial cells are the principal target of JCV productive infection in progressive multifocal leukoencephalopathy patients, little is known regarding the site of JCV persistence and the mechanisms by which the virus spreads to the CNS to cause disease. Previous work has demonstrated the presence of replicating JCV DNA in B lymphocytes from peripheral blood, tonsil, and spleen and it has been hypothesized that lymphocytes may be one site of JCV persistence. Detection of viral gene products in renal tubules and excretion of JC virions in the urine suggests JCV persistence in the kidney. A respiratory route of viral transmission has also been hypothesized implicating the lung as another possible site of persistent JCV infection. Earlier studies from our laboratory have shown that terminal alpha 2,6-linked sialic acid is a critical component of the JCV receptor. In this report we examined the tissue distribution of this JCV receptor-type sialic acid in a panel of normal human tissues. Our results demonstrate that in normal brain JCV receptor-type sialic acids are expressed on oligodendrocytes and astrocytes, but not on cortical neurons. The receptor-type sialic acid is also more highly expressed on B lymphocytes than on T lymphocytes in normal human spleen and tonsil. In addition, both the kidney and lung express abundant levels of alpha 2-6-linked sialic acids. Our data show a striking correlation between the expression of the JCV receptor-type sialic acid on cells and their susceptibility to infection by the virus. These findings also support the hypothesis of JCV persistence in lymphoid tissue and B-cell facilitated viral dissemination to the CNS. PMID- 14742249 TI - Differential expression of thromboxane synthase in prostate carcinoma: role in tumor cell motility. AB - Arachidonic acid metabolism through cyclooxygenase, lipoxygenase, or P-450 epoxygenase pathways can generate a variety of eicosanoids. Thromboxane synthase (TxS) metabolizes the cyclooxygenase product, prostanglandin H(2), into thromboxane A(2) (TXA(2)), which can cause vessel constriction, platelet activation, and aggregation. Here we demonstrate that human prostate cancer (PCa) cells express enzymatically active TxS and that this enzyme is involved in cell motility. In human PCa cell lines, PC-3, PC-3M, and ML-2 cells expressed higher levels of TxS than normal prostate epithelial cells or other established PCa cell lines such as DU145, LNCaP, or PPC-1. We cloned and sequenced the full-length TxS cDNA from PC-3 cells and found two changes in the amino acid residues. Immunohistochemical analysis of tumor specimens revealed that expression of TxS is weak or absent in normal differentiated luminal, or secretory cells, significantly elevated in less differentiated or advanced prostate tumors, and markedly increased in tumors with perineural invasion. TxS expressed in PC-3 cells was enzymatically active and susceptible to carboxyheptal imidazole, an inhibitor of TxS. The biosynthesis of TXA(2) in PC-3 cells was dependent on COX 2, and to a lesser extent, COX-1. Treatment of PC-3 cells with a COX-1 selective inhibitor, piroxicam, reduced TXA(2) synthesis by approximately 40%, while the COX-2 specific inhibitor NS398 reduced TXA(2) production by approximately 80%. Inhibition of TxS activity or blockade of TXA(2) function reduced PC-3 cell migration on fibronectin, while having minimal effects on cell cycle progression or survival. Finally, increased expression of TxS in DU145 cells increased cell motility. Our data suggest that human PCa cells express TxS and that this enzyme may contribute to PCa progression through modulating cell motility. PMID- 14742250 TI - Transplanted bone marrow cells repair heart tissue and reduce myocarditis in chronic chagasic mice. AB - A progressive destruction of the myocardium occurs in approximately 30% of Trypanosoma cruzi-infected individuals, causing chronic chagasic cardiomyopathy, a disease so far without effective treatment. Syngeneic bone marrow cell transplantation has been shown to cause repair and improvement of heart function in a number of studies in patients and animal models of ischemic cardiopathy. The effects of bone marrow transplant in a mouse model of chronic chagasic cardiomyopathy, in the presence of the disease causal agent, ie, the T. cruzi, are described herein. Bone marrow cells injected intravenously into chronic chagasic mice migrated to the heart and caused a significant reduction in the inflammatory infiltrates and in the interstitial fibrosis characteristics of chronic chagasic cardiomyopathy. The beneficial effects were observed up to 6 months after bone marrow cell transplantation. A massive apoptosis of myocardial inflammatory cells was observed after the therapy with bone marrow cells. Transplanted bone marrow cells obtained from chagasic mice and from normal mice had similar effects in terms of mediating chagasic heart repair. These results show that bone marrow cell transplantation is effective for treatment of chronic chagasic myocarditis and indicate that autologous bone marrow transplant may be used as an efficient therapy for patients with chronic chagasic cardiomyopathy. PMID- 14742251 TI - Increased expression of plasminogen activator inhibitor-1 in cardiomyocytes contributes to cardiac fibrosis after myocardial infarction. AB - Plasminogen activator inhibitor-1 (PAI-1) plays a critical role in tissue fibrosis by inactivating matrix metalloproteinases, which might effect on the progression of left ventricular dysfunction. However, little has been known about the expression of PAI-1 during cardiac remodeling. We used a mouse model of myocardial infarction (MI) by coronary ligation, in which the progression of left ventricular remodeling was confirmed by echocardiography. Histological examination showed that interstitial and perivascular fibrosis progressed in the post-MI (PMI) heart at 4 weeks after the procedure. We observed the dramatic induction of cardiac PAI-1 mRNA and PAI-1 antigen in plasma in the PMI mice, as compared with the sham-operated (sham) mice. In situ hybridization analysis demonstrated that strong signals for PAI-1 mRNA were localized to cardiomyocytes in the border of infarct area and around fibrous lesions, and to perivascular mononuclear cells, which seemed to be mast cells, only in hearts of the PMI mice. Importantly, less development of cardiac fibrosis after MI was observed in mice deficient in PAI-1 as compared to wild-type mice. The mRNA expression of cytokines, transforming growth factor-beta, and tumor necrosis factor-alpha, was also increased in hearts of the PMI mice, but not in the sham mice. These observations suggest that cardiomyocytes and mast cells contribute to the increased PAI-1 expression, resulting in the development of interstitial and perivascular fibrosis in the PMI heart, and that the regional induction of cytokines may be involved in this process. PMID- 14742252 TI - Impairment in ischemia-induced neovascularization in diabetes: bone marrow mononuclear cell dysfunction and therapeutic potential of placenta growth factor treatment. AB - Mechanisms that hinder ischemia-induced neovascularization in diabetes remain poorly understood. We hypothesized that endogenous bone marrow mononuclear cell (BM-MNC) dysfunction may contribute to the abrogated postischemic revascularization reaction associated with diabetes. We first analyzed the effect of diabetes (streptozotocin, 40 mg/kg) on BM-MNC pro-angiogenic potential in a model of surgically induced hindlimb ischemia. In nondiabetic animals, transplantation of BM-MNCs isolated from nondiabetic animals raised the ischemic/nonischemic angiographic score, capillary number, and blood flow recovery by 1.8-, 2.7-, and 2.2-fold, respectively, over that of PBS-injected nondiabetic animals (P < 0.05). Administration of diabetic BM-MNCs also improved the neovascularization reaction in ischemic hindlimbs of nondiabetic mice but to a lesser extent from that observed with nondiabetic BM-MNC transplantation. In diabetic mice, injection of nondiabetic BM-MNCs was still more efficient than that of diabetic BM-MNCs. Such BM-MNC dysfunction was associated with the impairment of diabetic BM-MNC capacity to differentiate into endothelial progenitor cells (EPCs) in vitro and to participate in vascular-like structure formation in a subcutaneous Matrigel plug. Placenta growth factor (PlGF) administration improved by sixfold the number of EPCs differentiated from diabetic BM-MNCs in vitro and enhanced ischemic/nonischemic angiographic score, capillary number and blood flow recovery by 1.9-, 1.5- and 1.6-fold, respectively, over that of untreated diabetic animals (P < 0.01). Endogenous BM MNC pro-angiogenic potential was affected in diabetes. Therapeutic strategy based on PlGF administration restored such defects and improved postischemic neovascularization in diabetic mice. PMID- 14742253 TI - Targeting the Tie2/Tek receptor in astrocytomas. AB - Tie2 is an endothelial cell-specific receptor tyrosine kinase, whose activation is positively and negatively modulated by angiopoietin-1 and angiopoietin-2, respectively. Angiopoietin-mediated modulation of Tie2 activation contributes to normal vessel development and stability, however, its role in tumor angiogenesis is not well known. We investigated the role of Tie2 activation in malignant astrocytomas, a common and highly vascularized primary human brain tumor. We found that Tie2 expression and activation increases with increasing malignancy grade of astrocytomas. Inhibition of Tie2, using a kinase-deficient Tie2 construct, decreases growth of malignant human astrocytoma subcutaneous and intracranial xenografts. Tie2 inactivation disrupted the tumor vascularity, with a decrease in microvascular density, increased presence of abnormally dilated vessels, and loss of interaction between endothelial cells and surrounding smooth muscle cells, all collectively resulting in increased tumor cell apoptosis. Overall, these findings strongly suggest that Tie2 activation contributes significantly to astrocytoma tumor angiogenesis and growth. We postulate that targeting Tie2 activation, either independently or in conjunction with other anti angiogenic therapies, such as against vascular endothelial growth factor, is of potential clinical interest. PMID- 14742254 TI - Inhibition of p38 mitogen-activated protein kinase augments progression of remnant kidney model by activating the ERK pathway. AB - p38, a mitogen-activated protein kinase, is a major intracellular signaling molecule involved in inflammation. To test the hypothesis that p38 mediates renal disease progression, we administered a novel p38 alpha inhibitor, NPC31169, to rats with remnant kidneys (RKs). RK rats showed increased p38 activation at 9 weeks (by p38 kinase assay), which was blocked by the inhibitor. In contrast to our expectation, treatment with the NPC31169 resulted in worse renal function, more proteinuria, and more severe glomerulosclerosis and tubulointerstitial injury. p38 inhibition resulted in marked cell proliferation in RK rats, with more proliferating tubular cells, myofibroblasts, and macrophages. In contrast, p38 suppression resulted in less tubular cell apoptosis. Interestingly, Western blot demonstrated increased ERK1/2 phosphorylation in p38-treated rats. No histological changes were observed in p38 inhibited sham-operated rats. Our findings indicate that, whereas blocking p38 usually shows benefit in inflammatory disease, in this model p38 inhibition resulted in accelerated renal progression. We conclude that blocking p38-dependent inflammation may have resulted in enhanced proliferation and increased ERK1/2 activation, and thereby explains the worse renal lesions observed. PMID- 14742255 TI - Chicken collagen X regulatory sequences restrict transgene expression to hypertrophic cartilage in mice. AB - Collagen X is produced by hypertrophic cartilage undergoing endochondral ossification. Transgenic mice expressing defective collagen X under the control of 4.7- or 1.6-kb chicken collagen X regulatory sequences yielded skeleto hematopoietic defects (Jacenko O, LuValle P, Olsen BR: Spondylometaphyseal dysplasia in mice carrying a dominant-negative mutation in a matrix protein specific for cartilage-to-bone transition. Nature 1993, 365:56-61; Jacenko O, Chan D, Franklin A, Ito S, Underhill CB, Bateman JF, Campbell MR: A dominant interference collagen X mutation disrupts hypertrophic chondrocyte pericellular matrix and glycosaminoglycan and proteoglycan distribution in transgenic mice. Am J Pathol 2001, 159:2257-2269; Jacenko O, Roberts DW, Campbell MR, McManus PM, Gress CJ, Tao Z: Linking hematopoiesis to endochondral ossification through analysis of mice transgenic for collagen X. Am J Pathol 2002, 160:2019-2034). Current data indicate that the hematopoietic abnormalities do not result from extraskeletal expression of endogenous collagen X or the transgene. Organs from mice carrying either promoter were screened by immunohistochemistry, in situ hybridization, and Northern blot; transgene and mouse collagen X proteins and messages were detected only in hypertrophic cartilage. Likewise, reverse transcriptase-polymerase chain reaction revealed both transgene and mouse collagen X amplicons only in the endochondral skeleton of mice with the 4.7-kb promoter; however, in mice with the 1.6-kb promoter, multiple organs were transgene-positive. Collagen X and transgene amplicons were also detected in marrow, but likely resulted from contaminating trabecular bone; this was supported by reverse transcriptase-polymerase chain reaction analysis of rat tibial zones free of trabeculae. Our data demonstrate that in mice, the 4.7-kb chicken collagen X promoter restricts transcription temporo-spatially to that of endogenous collagen X, and imply that murine skeleto-hematopoietic defects result from transgene co-expression with collagen X. Moreover, the 4.7-kb hypertrophic cartilage-specific promoter could be used for targeting transgenes to this tissue site in mice. PMID- 14742257 TI - Interleukin-10 secretion differentiates dendritic cells from human liver and skin. AB - Liver dendritic cells (DCs), which may orchestrate the liver's unique immunoregulatory functions, remain poorly characterized. We used a technique of overnight migration from pieces of normal human liver and skin to obtain tissue derived DCs with minimal culture and no additional cytokine treatment. Liver and skin DCs had a monocyte-like morphology and a partially mature phenotype, expressing myeloid markers, MHCII, and co-stimulatory molecules; but only the skin DCs contained a population of CD1a+ cells. Overnight-migrated liver DCs activated naive cord blood T cells efficiently. Liver DCs produced interleukin (IL)-10 whereas skin DCs failed to secrete IL-10 even after stimulation and neither skin nor liver-derived DCs secreted significant amounts of IL-12p70. Compared with skin DCs, liver DCs were less effective at stimulating T-cell proliferation and stimulated T cells to produce IL-10 and IL-4 whereas skin DCs were more potent stimulators of interferon-gamma and IL-4. Monocyte-derived DCs were down-regulated after culture with liver-conditioned media, suggesting that local microenvironmental factors may be important. Thus we show for the first time clear tissue-specific differences in nonlymphoid DCs. Although it is not possible to conclude from our data whether liver DCs are more regulatory, or skin DCs more proimmunogenic, the ability of liver DCs to secrete IL-10 may be important for regulating local immune responses within the liver in the face of constant exposure to gut antigens. PMID- 14742256 TI - Nuclear survivin expression in mantle cell lymphoma is associated with cell proliferation and survival. AB - Survivin is a member of the inhibitor of apoptosis protein family that is expressed in G2/M phase. Survivin is overexpressed and associated with parameters of poor prognosis in different human tumors. The role of survivin in the pathogenesis of mantle cell lymphoma (MCL) was examined in a series of typical and blastoid tumors. Survivin was detected as a nuclear pattern in a variable number of tumor cells. Mitotic figures were always positive with a strong delineation of the chromosomes. Western blot analysis confirmed the presence of survivin only in nuclear fractions. Protein expression detected by immunohistochemistry correlated with mRNA levels analyzed by quantitative real time reverse transcription-polymerase chain reaction (P < 0.0001). Survivin expression levels were higher in blastoid MCL variants (P < 0.0001) and were associated with the proliferative activity (P = 0.001), but not with the ploidy status of the tumors. The number of apoptotic cells was independent of survivin or Ki-67 expression. Overall survival was significantly shorter in patients with high survivin expression. However, in a multivariate analysis, proliferative index was a better predictor of survival than survivin score. These findings indicate that survivin is commonly expressed in MCL with a nuclear and mitotic pattern. The expression levels are strongly associated with the proliferative activity of the tumors and the survival of the patients, suggesting a potential role in cell cycle regulation and tumor progression. PMID- 14742258 TI - A peroxisome proliferator-activated receptor-alpha activator induces renal CYP2C23 activity and protects from angiotensin II-induced renal injury. AB - Cytochrome P450 (CYP)-dependent arachidonic acid (AA) metabolites are involved in the regulation of renal vascular tone and salt excretion. The epoxygenation product 11,12-epoxyeicosatrienoic acid (EET) is anti-inflammatory and inhibits nuclear factor-kappa B activation. We tested the hypothesis that the peroxisome proliferator-activated receptor-alpha-activator fenofibrate (Feno) induces CYP isoforms, AA hydroxylation, and epoxygenation activity, and protects against inflammatory organ damage. Double-transgenic rats (dTGRs) overexpressing human renin and angiotensinogen genes were treated with Feno. Feno normalized blood pressure, albuminuria, reduced nuclear factor-kappa B activity, and renal leukocyte infiltration. Renal epoxygenase activity was lower in dTGRs compared to nontransgenic rats. Feno strongly induced renal CYP2C23 protein and AA epoxygenase activity under pathological and nonpathological conditions. In both cases, CYP2C23 was the major isoform responsible for 11,12-EET formation. Moreover, we describe a novel CYP2C23-dependent pathway leading to hydroxy-EETs (HEETs), which may serve as endogenous peroxisome proliferator-activated receptor alpha activators. The capacity to produce HEETs via CYP2C23-dependent epoxygenation of 20-HETE and CYP4A-dependent hydroxylation of EETs was reduced in dTGR kidneys and induced by Feno. These results demonstrate that Feno protects against angiotensin II-induced renal damage and acts as inducer of CYP2C23 mediated epoxygenase activities. We propose that CYP-dependent EET/HEET production may serve as an anti-inflammatory control mechanism. PMID- 14742259 TI - Site-specific expression of polycomb-group genes encoding the HPC-HPH/PRC1 complex in clinically defined primary nodal and cutaneous large B-cell lymphomas. AB - Polycomb-group (PcG) genes preserve cell identity by gene silencing, and contribute to regulation of lymphopoiesis and malignant transformation. We show that primary nodal large B-cell lymphomas (LBCLs), and secondary cutaneous deposits from such lymphomas, abnormally express the BMI-1, RING1, and HPH1 PcG genes in cycling neoplastic cells. By contrast, tumor cells in primary cutaneous LBCLs lacked BMI-1 expression, whereas RING1 was variably detected. Lack of BMI-1 expression was characteristic for primary cutaneous LBCLs, because other primary extranodal LBCLs originating from brain, testes, and stomach were BMI-1-positive. Expression of HPH1 was rarely detected in primary cutaneous LBCLs of the head or trunk and abundant in primary cutaneous LBCLs of the legs, which fits well with its earlier recognition as a distinct clinical pathological entity with different clinical behavior. We conclude that clinically defined subclasses of primary LBCLs display site-specific abnormal expression patterns of PcG genes of the HPC HPH/PRC1 PcG complex. Some of these patterns (such as the expression profile of BMI-1) may be diagnostically relevant. We propose that distinct expression profiles of PcG genes results in abnormal formation of HPC-HPH/PRC1 PcG complexes, and that this contributes to lymphomagenesis and different clinical behavior of clinically defined LBCLs. PMID- 14742260 TI - Repair of local bone erosions and reversal of systemic bone loss upon therapy with anti-tumor necrosis factor in combination with osteoprotegerin or parathyroid hormone in tumor necrosis factor-mediated arthritis. AB - Local bone erosion and systemic bone loss are hallmarks of rheumatoid arthritis and cause progressive disability. Tumor necrosis factor (TNF) is a key mediator of arthritis and acts catabolically on bone by stimulating bone resorption and inhibiting bone formation. We hypothesized that the concerted action of anti-TNF, which reduces inflammation and parathyroid hormone (PTH), which stimulates bone formation, or osteoprotegerin (OPG), which blocks bone resorption and could lead to repair of local bone erosions and reversal of systemic bone loss. To test this, human TNF-transgenic mice with established erosive arthritis and systemic bone loss were treated with PTH, OPG, and anti-TNF, alone or in combination. Local bone erosions almost fully regressed, on combined treatment with anti-TNF and PTH and/or OPG, suggesting repair of inflammatory skeletal lesions. In contrast, OPG and anti-TNF alone led to arrest of bone erosions but did not achieve repair. Treatment with PTH alone had no influence on the progression of bone erosions. Local bone erosions all showed signs of new bone formation such as the presence of osteoblasts, osteoid formation, and mineralization. Furthermore, systemic bone loss was completely reversed on combined treatment and this effect was mediated by osteoblast stimulation and osteoclast blockade. In summary, we conclude that local joint destruction and systemic inflammatory bone loss because of TNF can regress and that repair requires a combined approach by reducing inflammation, blocking bone resorption, or stimulating bone formation. PMID- 14742261 TI - Up-regulation of the redox mediators thioredoxin and apurinic/apyrimidinic excision (APE)/Ref-1 in hypoxic microregions of invasive cervical carcinomas, mapped using multispectral, wide-field fluorescence image analysis. AB - Thioredoxin and apurinic/apyrimidinic excision (APE)/ref-1 are important redox mediators in biochemical pathways that promote cell survival under adverse conditions including hypoxia and oxidative stress. For example, elevated levels occur surrounding vascular infarcts and protect from reperfusion injury. Because elevated thioredoxin or APE/ref-1 is also associated with resistance to certain forms of cancer treatment, we examined their tissue distribution in a series of 110 cervical carcinoma biopsies. Analysis was done using a quadruple fluorescence imaging technique, incorporating carbonic anhydrase IX (CAIX) immunofluorescence to outline hypoxic microregions and 4',6-diamidino-2-phenylindole to localize nuclear staining of thioredoxin and APE/ref-1. A scanning autostage was used to image the entire tissue section. Thioredoxin and APE/ref-1 levels were expressed as the average pixel brightness in tumor tissue, subdivided based on CAIX and 4',6-diamidino-2-phenylindole staining. Results showed that the nuclear and cytoplasmic levels of thioredoxin were similar, whereas APE/ref-1 expression was greater in nuclei. Neither of these markers was predictive of outcome in this series of patients treated with radical radiotherapy. Both proteins showed highly significant elevations in CAIX-positive regions compared to CAIX-negative regions, and there was a nonsignificant trend for this effect to be greater in adenocarcinomas compared to squamous cell carcinomas. Levels of APE/ref-1 decreased with increasing tumor grade, but the expression was similar in CAIX positive regions of poorly differentiated tumors compared to moderately or well differentiated tumors. Elevated expression of thioredoxin and APE/ref-1 might promote cancer cell survival in hypoxic microenvironments of cervical carcinomas. PMID- 14742262 TI - Effect of osteopontin alleles on beta-glucan-induced granuloma formation in the mouse liver. AB - The granuloma formation is a host defense response against persistent irritants. Osteopontin is centrally involved in the formation of granulomas. Three osteopontin alleles, designated a, b, and c, have been found in mice. Here we used a murine model of zymosan (beta-glucan)-induced granuloma formation in the liver to determine possible functional differences between the osteopontin alleles in cell-mediated immunity. In contrast to mice with alleles a or c, mice with the allele b was defective in granuloma formation. As detected by mRNA expression, cytokines and chemokines known to be critically involved in granuloma formation were elicited in liver tissue, regardless of the osteopontin allele expressed. Alignment of the deduced amino acid sequences showed that unlike osteopontin c, b differs from a in 11 amino acids. All three osteopontin alleles had normal cell-binding properties. However, only the b allelic form was defective in the induction of cell migration as tested with dendritic cells. In conclusion, generation of a granulomatous response in mice depends critically on the presence of a functional osteopontin allele. Defective granuloma formation in mice with allele b is likely to be because of an impaired chemotactic function of the osteopontin b protein on immunocompetent cells. PMID- 14742263 TI - Basal cells are a multipotent progenitor capable of renewing the bronchial epithelium. AB - Commitment of the pulmonary epithelium to bronchial and bronchiolar airway lineages occurs during the transition from pseudoglandular to cannalicular phases of lung development, suggesting that regional differences exist with respect to the identity of stem and progenitor cells that contribute to epithelial maintenance in adulthood. We previously defined a critical role for Clara cell secretory protein-expressing (CE) cells in renewal of bronchiolar airway epithelium following injury. Even though CE cells are also the principal progenitor for maintenance of the bronchial airway epithelium, CE cell injury is resolved through a mechanism involving recruitment of a second progenitor cell population that we now identify as a GSI-B(4) reactive, cytokeratin-14-expressing basal cell. These cells exhibit multipotent differentiation capacity as assessed by analysis of cellular phenotype within clones of LacZ-tagged cells. Clones were derived from K14-expressing cells tagged in a cell-type-specific fashion by ligand-regulable Cre recombinase-mediated genomic rearrangement of the ROSA26 recombination substrate allele. We conclude that basal cells represent an alternative multipotent progenitor cell population of bronchial airways and that progenitor cell selection is dictated by the type of airway injury. PMID- 14742264 TI - Enhanced cytomegalovirus infection in atherosclerotic human blood vessels. AB - Human cytomegalovirus (CMV) is a possible co-factor in atherogenesis and vascular occlusion, but its ability to actively infect medium and large blood vessels is unclear. A vascular explant model was adapted to investigate CMV infection in human coronary artery, internal mammary artery (IMA), and saphenous vein (SV). Vascular explants were inoculated with CMV Towne or low-passage clinical isolate and examined in situ for CMV cytopathic effect and immediate-early and early antigens, as indicators of active infection. At 5 to 7 days after inoculation, we found that CMV Towne actively infected eight of eight different atherosclerotic blood vessel explants (coronary artery, n = 4; SV and IMA grafts, n = 4), whereas it only infected 2 of 14 nonatherosclerotic blood vessel explants (SV, n = 10; IMA, n = 4) (P = 0.001). The CMV clinical isolate actively infected none of six sets of nonatherosclerotic SV explants at 5 to 7 days after inoculation. The active CMV infections involved adventitial and, less frequently, intimal cells. A small subset of infected cells in atherosclerotic tissue expresses the endothelial cell marker CD31. Smooth muscle cells residing in both atherosclerotic and nonatherosclerotic blood vessels were free of active CMV infections even after all vascular tissue layers were exposed to the virus. In contrast, active CMV Towne infection was evident at 2 days after inoculation in smooth muscle cells and endothelial cells previously isolated from the SV tissues. We conclude that active CMV infection is enhanced in atherosclerotic blood vessels compared to atherosclerosis-free vascular equivalents, and this viral activity is restricted to subpopulations of intimal and adventitial cells. PMID- 14742265 TI - Smooth muscle hypertrophy following partial bladder outlet obstruction is associated with overexpression of non-muscle caldesmon. AB - Partial bladder outlet obstruction (PBOO) induces remodeling of urinary bladder smooth muscle (detrusor). We demonstrate an increase in bladder wall mass, muscle bundle size, and a threefold increase in the cross-sectional area of detrusor myocytes following PBOO in male New Zealand White rabbits compared to that of controls. Some bladders with detrusor hypertrophy function close to normal (compensated), whereas others were dysfunctional (decompensated), showing high intravesical pressure, large residual urine volume, and voiding difficulty. We analyzed the expression of smooth muscle-specific caldesmon (h-CaD) and non muscle (l-CaD) by Western blotting, RT-PCR, and real-time PCR. The expression of l-CaD is increased significantly at the mRNA and protein levels in the decompensated bladders compared to that of normal and compensated bladders. The CaD was also co-localized with myosin containing cytoplasmic fibrils in cells dissociated from obstructed bladders and cultured overnight. Our data show that the inability of decompensated bladders to empty, despite detrusor hypertrophy, is associated with an overexpression of l-CaD. The level of l-CaD overexpression might be a useful marker to estimate the degree of detrusor remodeling and contractile dysfunction in PBOO. PMID- 14742266 TI - Building an outcome predictor model for diffuse large B-cell lymphoma. AB - Diffuse large B-cell lymphoma (DLBCL) patients are treated using relatively homogeneous protocols, irrespective of their biological and clinical variability. Here we have developed a protein-expression-based outcome predictor for DLBCL. Using tissue microarrays (TMAs), we have analyzed the expression of 52 selected molecules in a series of 152 DLBCLs. The study yielded relevant information concerning key biological aspects of this tumor, such as cell-cycle control and apoptosis. A biological predictor was built with a training group of 103 patients, and was validated with a blind set of 49 patients. The predictive model with 8 markers can identify the probability of failure for a given patient with 78% accuracy. After stratifying patients according to the predicted response under the logistic model, 92.3% patients below the 25 percentile were accurately predicted by this biological score as "failure-free" while 96.2% of those above the 75 percentile were correctly predicted as belonging to the "fatal or refractory disease" group. Combining this biological score and the International Prognostic Index (IPI) improves the capacity for predicting failure and survival. This predictor was then validated in the independent group. The protein expression-based score complements the information obtained from the use of the IPI, allowing patients to be assigned to different risk categories. PMID- 14742267 TI - Collapse and restoration of MHC class-I-dependent immune privilege: exploiting the human hair follicle as a model. AB - The collapse of major histocompatibility complex (MHC) class-I-dependent immune privilege can lead to autoimmune disease or fetal rejection. Pragmatic and instructive models are needed to clarify the as yet obscure controls of MHC class I down-regulation in situ, to dissect the principles of immune privilege generation, maintenance, and collapse as well as to develop more effective strategies for immune privilege restoration. Here, we propose that human scalp hair follicles, which are abundantly available and easily studied, are ideally suited for this purpose: interferon-gamma induces ectopic MHC class I expression in the constitutively MHC class-I-negative hair matrix epithelium of organ cultured anagen hair bulbs, likely via interferon regulatory factor-1, along with up-regulation of the MHC class I pathway molecules beta(2)microglobulin and transporter associated with antigen processing (TAP-2). In the first report to identify natural immunomodulators capable of down-regulating MHC class I expression in situ in a normal, neuroectoderm-derived human tissue, we show that ectopic MHC class I expression in human anagen hair bulbs can be normalized by treatment with alpha-MSH, IGF-1, or TGF-beta1, all of which are locally generated, as well as by FK506. These agents are promising candidates for immune privilege restoration and for suppressing MHC class I expression where this is clinically desired (eg, in alopecia areata, multiple sclerosis, autoimmune uveitis, mumps orchitis, and fetal or allograft rejection). PMID- 14742268 TI - CXCR3 is involved in tubulointerstitial injury in human glomerulonephritis. AB - Chemokines play pivotal roles in the recruitment of inflammatory cells into the kidney. The chemokine receptors CXCR3 and CCR5 are expressed on activated T lymphocytes, and expression of CXCR3 by mesangial cells has been suggested. Detailed description of CXCR3 expression might form a rational basis for use as a diagnostic marker and for therapeutic CXCR3 targeting in human glomerulonephritis. We studied the expression of CXCR3 in renal biopsies by immunohistochemistry (n = 45), and real time RT-PCR (n = 78). Biopsies were from patients with IgA nephropathy, lupus nephritis, and membranoproliferative glomerulonephritis. Furthermore, cultured human mesangial cells (HMC) were studied for CXCR3 expression, and for functional responses to the ligands CXCL10/IP-10 and CXCL9/Mig. CXCR3-positive cells were rarely found in glomerular tufts, but formed a major part of the tubulointerstitial infiltrates. Consistently, CXCR3 mRNA expression was too low to be quantified in glomerular compartments, and was not detectable in HMC. The published staining for CXCR3 of mesangial cells could be traced to cross-reactivity of an antibody for CXCR3 with a potentially related chemokine receptor as revealed by FACS analysis. Despite an absence of CXCR3 expression, mesangial cells reacted to CXCR3 ligands by proliferation and migration, which was blocked by pertussis toxin but not by an anti-CXCR3 antibody. These results indicate that HMC do not express the classical CXCR3, but may potentially express a related receptor with shared ligand specificity. By immunohistochemistry the number of CXCR3-positive cells, mainly interstitial T cells, correlated with renal function, proteinuria, and percentage of globally sclerosed glomeruli. A significant morphological and numerical correlation between CD3, CXCR3, and CCR5-positive cells indicated a CXCR3/CCR5 double-positive T cell population. No apparent difference in the CXCR3 expression pattern was found between disease entities. CXCR3 expression was localized to interstitial T cells, and these cells correlated strongly with important prognostic markers. Therefore interstitial CXCR3, as well as CCR5-positive T cells might play an important role during progressive loss of renal function, and are potential therapeutic targets in human glomerular diseases. PMID- 14742269 TI - Smad3 signaling is required for epithelial-mesenchymal transition of lens epithelium after injury. AB - Lens epithelial cells undergo epithelial-mesenchymal transition (EMT) after injury as in cataract extraction, leading to fibrosis of the lens capsule. Fibrosis of the anterior capsule can be modeled in the mouse by capsular injury in the lens, which results in EMT of the lens epithelium and subsequent deposition of extracellular matrix without contamination of other cell types from outside the lens. We have previously shown that signaling via Smad3, a key signal transducing element downstream of transforming growth factor (TGF)-beta and activin receptors, is activated in lens epithelial cells by 12 hours after injury and that this Smad3 activation is blocked by administration of a TGF-beta 2 neutralizing antibody in mice. We now show that EMT of primary lens epithelial cells in vitro depends on TGF-beta expression and that injury-induced EMT in vivo depends, more specifically, on signaling via Smad3. Loss of Smad3 in mice blocks both morphological changes of lens epithelium to a mesenchymal phenotype and expression of the EMT markers snail, alpha-smooth muscle actin, lumican, and type I collagen in response to injury in vivo or to exposure to exogenous TGF-beta in organ culture. The results suggest that blocking the Smad3 pathway might be beneficial in inhibiting capsular fibrosis after injury and/or surgery. PMID- 14742270 TI - Of mice and dogs: species-specific differences in the inflammatory response following myocardial infarction. AB - Large animal models have provided much of the descriptive data regarding the cellular and molecular events in myocardial infarction and repair. The availability of genetically altered mice may provide a valuable tool for specific cellular and molecular dissection of these processes. In this report we compare closed chest models of canine and mouse infarction/reperfusion qualitatively and quantitatively for temporal, cellular, and spatial differences. Much like the canine model, reperfused mouse hearts are associated with marked induction of endothelial adhesion molecules, cytokines, and chemokines. Reperfused mouse infarcts show accelerated replacement of cardiomyocytes by granulation tissue leading to a thin mature scar at 14 days, when the canine infarction is still cellular and evolving. Infarcted mouse hearts demonstrate a robust but transient postreperfusion inflammatory reaction, associated with a rapid up-regulation of interleukin-10 and transforming growth factor-beta. Unlike canine infarcts, infarcted mouse hearts show only transient macrophage infiltration and no significant mast cell accumulation. In correlation, the growth factor for macrophages, M-CSF, shows modest and transient up-regulation in the early days of reperfusion; and the obligate growth factor for mast cells, stem cell factor, SCF, is not induced. In summary, the postinfarction inflammatory response and resultant repair in the mouse heart shares many common characteristics with large mammalian species, but has distinct temporal and qualitative features. These important species-specific differences should be considered when interpreting findings derived from studies using genetically altered mice. PMID- 14742271 TI - The impact of gender on progression of renal disease: potential role of estrogen mediated vascular endothelial growth factor regulation and vascular protection. AB - Male gender is associated with a more rapid progression of renal disease independent of blood pressure, dietary protein intake, or serum lipid levels. Recently, we reported a key role for the intrarenal vasculature in progressive renal disease (Kang D-H, Kanellis J, Hugo C, Truong L, Anderson S, Kerjaschki D, Schreiner GF, Johnson RJ: Role of endothelium in progressive renal disease. J Am Soc Nephrol 2002, 13:806-816). We hypothesized that estrogen-mediated preservation of the renal vasculature could account for the better renal outcome in female rats. We analyzed micro- and macrovascular changes in the 5/6 remnant kidney (RK) models both in male (n = 24) and female (n = 24) Sprague-Dawley rats up to 12 weeks after renal mass reduction. At 12 weeks, male and female RK rats had equivalent blood pressure, glomerular tuft area, and RK/body weight, but male rats showed worse renal function, proteinuria, glomerulosclerosis (%), and tubulointerstitial fibrosis. At 12 weeks peritubular capillary (PTC) EC proliferation and PTC density were higher in female RK rats whereas macrovascular changes in preglomerular vessels (smooth muscle cell proliferation, medial wall thickening, and adventitial fibrosis) were less prominent. The expression of vascular endothelial growth factor (VEGF) and VEGF type 2 receptor (flk-1) in renal cortex assessed by immunostaining were higher in female RK rats. To dissect the mechanism of sex hormone-induced vascular remodeling and VEGF regulation, we investigated the in vitro effect of 17 beta-estradiol (17 beta E, 10 nmol/L) on proliferation and VEGF expression of renal tubular cells (rat proximal tubular cells), vascular smooth muscle cells (VSMCs), and human umbilical vein endothelial cells (HUVECs). 17 beta E directly stimulated the proliferation of HUVECs, whereas it inhibited serum-induced proliferation of VSMCs. 17 beta E stimulated VEGF mRNA expression both in renal tubular cells and VSMCs. However, when cells were pretreated with a nitric oxide donor to simulate the in vivo condition, 17 beta E inhibited VEGF mRNA expression and protein release in VSMCs. In conclusion, female RK rats developed less glomerulosclerosis and renal failure compared to male RK rats in association with greater preservation of PTC and less preglomerular arteriopathy. Estrogen stimulated basal VEGF expression in renal tubular cells. We propose that estrogen may protect female rats in progressive renal disease by stimulating VEGF expression and maintaining a healthy intrarenal vasculature. PMID- 14742272 TI - Increased DNA methyltransferase 1 (DNMT1) protein expression correlates significantly with poorer tumor differentiation and frequent DNA hypermethylation of multiple CpG islands in gastric cancers. AB - We evaluated the significance of aberrant DNA methyltransferase 1 (DNMT1) protein expression during gastric carcinogenesis. The protein expression of DNMT1, Muc2, human gastric mucin, E-cadherin, and proliferating cell nuclear antigen was examined immunohistochemically in gastric cancers and corresponding noncancerous mucosae from 134 patients. The DNA methylation status of the CpG islands of the p16, human MutL homologue 1 (hMLH1), E-cadherin, and thrombospondin-1 (THBS-1) genes and the methylated in tumor (MINT)-1, -2, -12, and -31 clones was examined by methylation-specific polymerase chain reaction and combined bisulfite restriction enzyme analysis. Epstein-Barr virus (EBV) infection was detected by in situ hybridization. Nuclear immunoreactivity for DNMT1 was not detected in any of the noncancerous epithelia, except in proliferative zones (positive internal control), but was found in 97 (72%) of the gastric cancers. DNMT1 overexpression correlated significantly with poorer tumor differentiation (P < 0.001), but not with the phenotype (gastric type versus intestinal type) of the cancer cells. It also correlated significantly with DNA hypermethylation of the CpG islands of the hMLH1 (P = 0.024) and THBS-1 genes (P = 0.043), and with the CpG island methylator phenotype in the gastric cancers (P = 0.007). Reduced E-cadherin expression correlated significantly with poorer tumor differentiation (P = 0.002), DNA hypermethylation of the E-cadherin gene (P < 0.001) and DNMT1 overexpression (P = 0.014). DNMT1 overexpression was also associated with EBV infection (a potential etiological factor in gastric carcinogenesis) but not with the proliferative activity of the cancer cells as indicated by the proliferating cell nuclear antigen-labeling index. These results suggest that DNMT1 overexpression may not be just a secondary effect of increased cancer cell proliferative activity, but may be associated with EBV infection and other etiological factors during gastric carcinogenesis. Furthermore, DNMT1 may play a significant role in the development of poorly differentiated gastric cancers by inducing frequent DNA hypermethylation of multiple CpG islands. PMID- 14742273 TI - Retinal dystrophy resulting from ablation of RXR alpha in the mouse retinal pigment epithelium. AB - Vitamin A (retinol) actions in eye development are mediated by retinoic acid receptors (RARs and RXRs). Using the Cre/loxP system, we have selectively ablated RXR alpha in the retinal pigment epithelium (RPE), a cell monolayer critically involved in visual retinoid renewal and phagocytosis of photoreceptor outer segments. In the mutant (RXR alpha (rpe-/-)) mice, RPE cells are morphologically and functionally abnormal and display decreased expression of proteins involved in the visual retinoid cycle, namely RPE65, CRALBP, and RGR. RXR alpha (rpe-/-) mice also show alterations of photoreceptor cells including: 1) decrease in their number; 2) outer segment shortening and disorganization, and 3) reduced light responses in electroretinograms. These results indicate that RXR alpha is required for normal maturation of the RPE, which is known to play essential roles in photoreceptor cell function and survival, and point to a possible involvement of RXR alpha signaling pathways in the RPE in human retinal diseases. PMID- 14742274 TI - Transgenic overexpression of dystroglycan does not inhibit muscular dystrophy in mdx mice. AB - Recently, there have been a number of studies demonstrating that overexpression of molecules in skeletal muscle can inhibit or ameliorate aspects of muscular dystrophy in the mdx mouse, a model for Duchenne muscular dystrophy. Several such studies involve molecules that increase the expression of dystroglycan, an important component of the dystrophin-glycoprotein complex. To test whether dystroglycan itself inhibits muscular dystrophy in mdx mice, we created dystroglycan transgenic mdx mice (DG/mdx). The alpha and beta chains of dystroglycan were highly overexpressed along the sarcolemmal membrane in most DG/mdx muscles. Increased dystroglycan expression, however, did not correlate with increased expression of utrophin or sarcoglycans, but rather caused their decreased expression. In addition, the percentage of centrally located myofiber nuclei and the level of serum creatine kinase activity were not decreased in DG/mdx mice relative to mdx animals. Therefore, dystroglycan overexpression does not cause the concomitant overexpression of a utrophin-glycoprotein complex in mdx muscles and has no effect on the development of muscle pathology associated with muscular dystrophy. PMID- 14742275 TI - Beta-secretase activity increases with aging in human, monkey, and mouse brain. AB - Amyloid beta protein (A beta) accumulates in the brains of aging humans, amyloid precursor protein (APP) transgenic mouse lines, and rhesus monkeys. We tested the hypothesis that aging was associated with increased activity of the beta-site amyloid precursor protein cleaving enzyme (beta-secretase, BACE) in brain. We evaluated BACE activity, BACE protein, and formic acid-extractable A beta levels in cohorts of young (4 months old) and old (14 to 18 months old) nontransgenic mice (n = 16) and Tg2576 APP transgenic mice (n = 17), young (4.4 to 12.7 years old) and old (20.9 to 30.4 years old) rhesus monkeys (n = 17), and a wide age range (18 to 92 years old) of nondemented human brains (n = 25). Aging was associated with increased brain A beta levels in each cohort. Furthermore BACE activity increased significantly with age in mouse, monkey, and human brains, independent of brain region. BACE protein levels, however, were unchanged with age. BACE activity correlated with formic acid-extractable A beta levels in transgenic mouse, nontransgenic mouse, and human cortex, but not in monkey brain. These data suggest that an age-related increase of BACE activity contributes to the increased production and accumulation of brain A beta, and potentially predisposes to Alzheimer's disease in humans. PMID- 14742276 TI - Abnormalities in alpha-dystroglycan expression in MDC1C and LGMD2I muscular dystrophies. AB - We recently identified mutations in the fukutin related protein (FKRP) gene in patients with congenital muscular dystrophy type 1C (MDC1C) and limb girdle muscular dystrophy type 2I (LGMD2I). The sarcolemma of these patients typically displays an immunocytochemical reduction of alpha-dystroglycan. In this report we extend these observations and report a clear correlation between the residual expression of alpha-dystroglycan and the phenotype. Three broad categories were identified. Patients at the severe end of the clinical spectrum (MDC1C) were compound heterozygote between a null allele and a missense mutation or carried two missense mutations and displayed a profound depletion of alpha-dystroglycan. Patients with LGMD with a Duchenne-like severity typically had a moderate reduction in alpha-dystroglycan and were compound heterozygotes between a common C826A (Leu276Ileu) FKRP mutation and either a missense or a nonsense mutation. Individuals with the milder form of LGMD2I were almost invariably homozygous for the Leu276Ile FKRP mutation and showed a variable but subtle alteration in alpha dystroglycan immunolabeling. Our data therefore suggest a correlation between a reduction in alpha-dystroglycan, the mutation and the clinical phenotype in MDC1C and LGMD2I which supports the hypothesis that dystroglycan plays a central role in the pathogenesis of these disorders. PMID- 14742277 TI - An engineered biopolymer prevents mucositis induced by 5-fluorouracil in hamsters. AB - Oral mucositis is a common, treatment-limiting, and costly side effect of cancer treatments whose biological underpinnings remain poorly understood. In this study, mucositis induced in hamsters by 5-fluorouracil (5-FU) was observed after cheek-pouch scarifications, with and without administration of RGTA (RG1503), a polymer engineered to mimic the protective effects of heparan sulfate. RG1503 had no effects on 5-FU-induced decreases in body weight, blood cell counts, or cheek pouch and jejunum epithelium proliferation rates, suggesting absence of interference with the cytotoxic effects of 5-FU. Extensive mucositis occurred in all of the untreated animals, and consisted of severe damage to cheek pouch tissues (epithelium, underlying connective tissue, and muscle bundles). Only half of the RG1503-treated animals had mucositis, over a mean area 70% smaller than in the untreated animals. Basement membranes were almost completely destroyed in the untreated group but was preserved in the RG1503 group. RG1503 blunted or abolished the following 5-FU-induced effects: increases in matrix metalloproteinase (MMP)-2, MMP-9, and plasmin, and decreases in tissue inhibitor of metalloproteinase (TIMP)-1 and TIMP-2. These data indicate that mucositis lesions are related to massive release of proteolytic enzymes and are improved by RG1503 treatment, this effect being ascribable in part to restoration of the MMP TIMP balance. RG1503 given with cancer treatment might protect patients from mucositis. PMID- 14742278 TI - Nonirradiated NOD/SCID-human chimeric animal model for primary human multiple myeloma: a potential in vivo culture system. AB - The NOD/SCID human chimeric animal model was generated by implanting of human fetal bones (FBs) into subcutaneous sites of NOD/SCID mice (NOD/SCID-hu(+)), followed by inoculation of primary bone marrow mononuclear cells (BMNCs) obtained from patients with multiple myeloma (MM) into the FBs. The BMNCs from 30 patients with MM were inoculated, and 28 (93%) of them revealed evidence of tumor growth of myeloma cells (MCs) in the NOD/SCID-hu(+) mice. Intriguingly, 17 (61%) of the 28 patients' BMNCs inoculated developed not only myeloma in the bone marrow of the FBs, but also extramedullary macrotumors (EMTs) along the periosteum of the FBs. The tumor cells in these EMTs had plasmacytoid morphology and preserved antigens and cytogenetics similar, if not identical, to those in the parent MCs. Moreover, small tumor blocks from nine EMTs were transplanted into subcutaneous sites of subsequent recipient NOD/SCID mice without human FBs (NOD/SCID-hu(-)), and all but one grew successfully. Two of the EMTs have been maintained in the animal model for more than 12 months. The NOD/SCID-hu(+) chimeric animal model is highly efficient for growth of primary MCs and presents clinical features of human MM. The engrafted MCs can be maintained subsequently in NOD/SCID-hu(-) mice as in vivo culture. PMID- 14742279 TI - A generalized linear mixed models approach for detecting incident clusters of disease in small areas, with an application to biological terrorism. AB - Since the intentional dissemination of anthrax through the US postal system in the fall of 2001, there has been increased interest in surveillance for detection of biological terrorism. More generally, this could be described as the detection of incident disease clusters. In addition, the advent of affordable and quick geocoding allows for surveillance on a finer spatial scale than has been possible in the past. Surveillance for incident clusters of disease in both time and space is a relatively undeveloped arena of statistical methodology. Surveillance for bioterrorism detection, in particular, raises unique issues with methodological relevance. For example, the bioterrorism agents of greatest concern cause initial symptoms that may be difficult to distinguish from those of naturally occurring disease. In this paper, the authors propose a general approach to evaluating whether observed counts in relatively small areas are larger than would be expected on the basis of a history of naturally occurring disease. They implement the approach using generalized linear mixed models. The approach is illustrated using data on health-care visits (1996-1999) from a large Massachusetts managed care organization/multispecialty practice group in the context of syndromic surveillance for anthrax. The authors argue that there is great value in using the geographic data. PMID- 14742280 TI - Invited commentary: Surveilling surveillance--some statistical comments. PMID- 14742282 TI - Do men have a higher case fatality rate of severe acute respiratory syndrome than women do? AB - Severe acute respiratory syndrome (SARS) has been reported in 30 countries and regions, with a cumulative total of 8,099 probable cases and 774 deaths as of July 31, 2003, according to the World Health Organization. In Hong Kong, People's Republic of China, 1,755 SARS cases and 299 deaths had occurred as of September 22, 2003. The authors analyzed data from the Department of Health, Hong Kong SAR. The data series includes details regarding sex, age, and chronic disease history. Using data from early March to September 22, 2003, the authors found that males had a significantly (p < 0.0001) higher case fatality rate than females did, 21.9% versus 13.2%; the relative risk was 1.66 (95% confidence interval (CI): 1.35, 2.05), and it was 1.62 (95% CI: 1.21, 2.16) after adjustment for age. Subgroup analysis was conducted by excluding health care workers (n = 386) from the analysis. The overall crude relative risk of mortality was 1.41 (95% CI: 1.15, 1.74), and the adjusted relative risk was 1.48 (95% CI: 1.10, 2.00). Thus, among SARS patients, males may be more severely affected by the disease than females are. This finding could be related to a nonuniform case definition of SARS disease, a different treatment regimen, a past smoking history, work environment factors, or gender-specific immune-defense factors, for instance. PMID- 14742283 TI - Fraction of cases of acquired immunodeficiency syndrome prevented by the interactions of identified restriction gene variants. AB - Previous research has demonstrated isolated effects of host genetic factors on the progression of human immunodeficiency virus type 1 (HIV-1) infection. In this paper, the authors present a novel use of multivariable methods for estimating the prevented fraction of acquired immunodeficiency syndrome (AIDS) cases attributable to six restriction genes after accounting for their epidemiologic interactions. The methods presented will never yield a prevented fraction above 1. The study population consisted of a well-characterized cohort of 525 US men with HIV-1 seroconversion documented during follow-up (1984-1996). On the basis of a regression tree approach using a Cox proportional hazards model for times to clinical AIDS, the combinations of genes associated with the greatest protection, relative to the lack of a protective genotype, consisted of: 1) C-C chemokine receptor 5 (CCR5)-Delta 32 and C-C chemokine receptor 2 (CCR2)-64I (relative hazard = 0.44); 2) interleukin 10 (IL10)-+/+ in combination with CCR5-Delta 32 or CCR2-64I (relative hazard = 0.45); and 3) IL10-+/+ in combination with stromal derived factor (SDF1)-3 'A and CCR5 promoter P1/approximately P1 (relative hazard = 0.37). Overall, 30% of potential AIDS cases were prevented by the observed combinations of restriction genes (95% confidence interval: 7, 47). However, the combined effect was confined to the first 4 years following HIV-1 seroconversion. Additional research is needed to identify AIDS restriction genes with stronger and long-lasting protection to better characterize the genetic epidemiology of HIV-1. PMID- 14742284 TI - Condom effectiveness for reducing transmission of gonorrhea and chlamydia: the importance of assessing partner infection status. AB - This analysis examined the importance of differential exposure to infected partners in epidemiologic studies of latex condom effectiveness for prevention of sexually transmitted infections. Cross-sectional, enrollment visit data were analyzed from Project RESPECT, a trial of counseling interventions conducted at five publicly funded US sexually transmitted disease clinics between 1993 and 1997. The association between consistent condom use in the previous 3 months and prevalent gonorrhea and chlamydia (Gc/Ct) was compared between participants known to have infected partners and participants whose partner infection status was unknown. Among 429 participants with known Gc/Ct exposure, consistent condom use was associated with a significant reduction in prevalent gonorrhea and chlamydia (30% vs. 43%; adjusted prevalence odds ratio = 0.42, 95% confidence interval: 0.18, 0.99). Among 4,314 participants with unknown Gc/Ct exposure, consistent condom use was associated with a lower reduction in prevalent gonorrhea and chlamydia (24% vs. 25%; adjusted prevalence odds ratio = 0.82, 95% confidence interval: 0.66, 1.01). The number of unprotected sex acts was significantly associated with infection when exposure was known (p for trend < 0.01) but not when exposure was unknown (p for trend = 0.73). Restricting analyses to participants with known exposure to infected partners provides a feasible and efficient mechanism for reducing confounding from differential exposure to infected partners in condom effectiveness studies. PMID- 14742285 TI - Is Helicobacter pylori infection a necessary condition for noncardia gastric cancer? AB - Although the association between Helicobacter pylori infection and gastric cancer is well established, this association might have been underestimated in epidemiologic studies because of possible clearance of the infection in the course of disease development. The authors addressed this hypothesis in a case control study from Saarland, Germany (68 cases first diagnosed between 1996 and 1998 and 360 controls), with serologic assessment of H. pylori infection in which various exclusion criteria were used to minimize potential bias from this source. Joint application of three such exclusion criteria (blood sample taken more than 90 days after gastrectomy, advanced (T4) gastric cancer, and CagA positivity in Western blot analysis despite a negative result in anti-H. pylori immunoglobulin G enzyme-linked immunosorbent assay) increased the odds ratio of noncardia gastric cancer from 3.7 (95% confidence interval (CI): 1.7, 7.9) to 18.3 (95% CI: 2.4, 136.7) for any H. pylori infection and from 5.7 (95% CI: 2.6, 12.8) to 28.4 (95% CI: 3.7, 217.1) for CagA-positive H. pylori infections. Furthermore, there was no single H. pylori-negative patient out of 32 patients with noncardia gastric cancer left after additional exclusion of subjects with borderline levels in immunoglobulin G enzyme-linked immunosorbent assay. The H. pylori-gastric cancer relation may be much stronger than previously thought, and H. pylori infection may even be a (close to) necessary condition for development of noncardia gastric cancer. PMID- 14742286 TI - Association of obesity and cancer risk in Canada. AB - The authors conducted a population-based, case-control study of 21,022 incident cases of 19 types of cancer and 5,039 controls aged 20-76 years during 1994-1997 to examine the association between obesity and the risks of various cancers. Compared with people with a body mass index of less than 25 kg/m(2), obese (body mass index of > or = 30 kg/m(2)) men and women had an increased risk of overall cancer (multivariable adjusted odds ratio = 1.34, 95% confidence interval (CI): 1.22, 1.48), non-Hodgkin's lymphoma (odds ratio = 1.46, 95% CI: 1.24, 1.72), leukemia (odds ratio = 1.61, 95% CI: 1.32, 1.96), multiple myeloma (odds ratio = 2.06, 95% CI: 1.46, 2.89), and cancers of the kidney (odds ratio = 2.74, 95% CI: 2.30, 3.25), colon (odds ratio = 1.93, 95% CI: 1.61, 2.31), rectum (odds ratio = 1.65, 95% CI: 1.36, 2.00), pancreas (odds ratio = 1.51, 95% CI: 1.19, 1.92), breast (in postmenopausal women) (odds ratio = 1.66, 95% CI: 1.33, 2.06), ovary (odds ratio = 1.95, 95% CI: 1.44, 2.64), and prostate (odds ratio = 1.27, 95% CI: 1.09, 1.47). Overall, excess body mass accounted for 7.7% of all cancers in Canada-9.7% in men and 5.9% in women. This study provides further evidence that obesity increases the risk of overall cancer, non-Hodgkin's lymphoma, leukemia, multiple myeloma, and cancers of the kidney, colon, rectum, breast (in postmenopausal women), pancreas, ovary, and prostate. PMID- 14742287 TI - Androgen receptor gene polymorphisms and increased risk of urologic measures of benign prostatic hyperplasia. AB - The association between androgen receptor gene polymorphisms and benign prostatic hyperplasia was investigated among 510 men randomly selected from Olmsted County, Minnesota. From 1990 through 2000, lower urinary tract symptom severity was assessed by the American Urological Association Symptom Index (AUASI), and peak urinary flow rate, prostate volume, and serum prostate-specific antigen level were measured. Androgen receptor CAG and GGN genotyping was performed. A CAG repeat length of <21 was associated with an enlarged prostate (hazard ratio (HR) = 1.4, 95% confidence interval (CI): 1.0, 1.9) and a serum prostate-specific antigen level >1.4 ng/ml (HR = 1.5, 95% CI: 1.1, 2.0). A GGN repeat length of <16 was associated with an AUASI >7 (HR = 1.6, 95% CI: 1.1, 2.3) and a serum prostate specific antigen level >1.4 ng/ml (HR = 1.5, 95% CI: 1.0, 2.3). Having <21 CAG repeats and <16 GGN repeats compared with having neither was associated with an enlarged prostate (HR = 2.5, 95% CI: 1.5, 4.2), a serum prostate-specific antigen level >1.4 ng/ml (HR = 2.8, 95% CI: 1.6, 4.7), a peak flow rate <12 ml/second (HR = 1.9, 95% CI: 1.1, 3.4), and an AUASI >7 (HR = 1.6, 95% CI: 1.0, 2.7). Androgen receptor gene polymorphisms may have a potential role in the pathogenesis of benign prostatic hyperplasia. PMID- 14742288 TI - Cellular telephone use and risk of acoustic neuroma. AB - Despite limited evidence, cellular telephones have been claimed to cause cancer, especially in the brain. In this Danish study, the authors examined the possible association between use of cellular telephones and development of acoustic neuroma. Between 2000 and 2002, they ascertained 106 incident cases and matched these persons with 212 randomly sampled, population-based controls on age and sex. The data obtained included information on use of cellular telephones from personal interviews, data from medical records, and the results of radiologic examinations. The authors obtained information on socioeconomic factors from Statistics Denmark. The overall estimated relative risk of acoustic neuroma was 0.90 (95% confidence interval: 0.51, 1.57). Use of a cell phone for 10 years or more did not increase acoustic neuroma risk over that of short-term users. Furthermore, tumors did not occur more frequently on the side of the head on which the telephone was typically used, and the size of the tumor did not correlate with the pattern of cell phone use. The results of this prospective, population-based, nationwide study, which included a large number of long-term users of cellular telephones, do not support an association between cell phone use and risk of acoustic neuroma. PMID- 14742289 TI - A prospective study of microalbuminuria and incident coronary heart disease and its prognostic significance in a British population: the EPIC-Norfolk study. AB - Microalbuminuria is associated with an increased risk of cardiovascular and renal disease in patients with diabetes and hypertension. The role of microalbuminuria as a predictor of coronary heart disease (CHD) has not been examined in large general-population cohorts, and its prognostic significance in persons with established CHD is uncertain. The authors examined the relation between microalbuminuria and incident CHD (1993-2002) in a population-based British cohort of 22,368 men and women aged 40-79 years without prevalent baseline CHD and evaluated its prognostic significance in 1,596 participants with baseline CHD. Participants were members of the Norfolk, United Kingdom, component of the European Prospective Investigation into Cancer and Nutrition (the EPIC-Norfolk Study). At baseline, participants were categorized into normoalbuminuria, microalbuminuria, and macroalbuminuria groups. During an average of 6.4 years of follow-up, 800 primary CHD events were registered. The age-adjusted incidence of CHD increased significantly across ordered categories of albuminuria (4.3, 4.4, and 5.6/1,000 person-years across tertiles of normoalbuminuria, 7.1/1,000 person years for microalbuminuria, and 12.2/1,000 person-years for macroalbuminuria; p for trend < 0.001). The multivariate hazard ratio for incident primary CHD was 1.36 (95% confidence interval (CI): 1.12, 1.64) for microalbuminuria and 1.59 (95% CI: 1.10, 2.37) for macroalbuminuria. Among participants with established baseline CHD, the independent risk of all-cause mortality associated with microalbuminuria was 1.61 (95% CI: 1.19, 2.07). Microalbuminuria may be useful in identifying persons at increased risk of CHD and subsequent death in the general population. PMID- 14742290 TI - Gaseous air pollutants and asthma hospitalization of children with low household income in Vancouver, British Columbia, Canada. AB - Associations of gaseous air pollutants (including carbon monoxide, sulfur dioxide, nitrogen dioxide, and ozone) with asthma hospitalization, stratified by sex and socioeconomic status, were examined among children 6-12 years of age in Vancouver, British Columbia, Canada, between 1987 and 1998. Relative risks for an exposure increment corresponding to the interquartile range for each gaseous air pollutant were estimated for asthma hospitalization after adjustment for weather conditions, including daily maximum and minimum temperatures as well as average relative humidity. Similar results were obtained by using locally weighted smoothing functions (LOESS) with default convergence criteria and by using natural cubic splines with a more stringent setting. Exposures to nitrogen dioxide were found to be significantly and positively associated with asthma hospitalization for males in the low socioeconomic group but not in the high socioeconomic group. For females, this same pattern of association was observed for exposures to sulfur dioxide. No significantly positive associations were found between carbon monoxide and ozone and asthma hospitalization in either low or high socioeconomic groups. PMID- 14742291 TI - Recall of physical activity in the distant past: the 32-year follow-up of the Prospective Population Study of Women in Goteborg, Sweden. AB - It has been of interest to researchers in chronic disease epidemiology to determine whether elderly subjects can accurately recall their physical activity patterns in the distant past. As part of a 32-year follow-up of the Prospective Population Study of Women in Goteborg, Sweden, 433 participants currently aged 70 92 years recalled their leisure-time physical activity at the original examination in 1968, when they had been 38-60 years of age. Using a four-point scale, subjects were originally asked to describe leisure-time activity during the previous 12 months. Identical questions were asked in 2000 describing levels in 1968 and current levels. Subjects were more likely to overestimate their previous activity level than to underestimate it: 43.9% of women classified themselves consistently at both examinations; 48.7% overestimated and 7.4% underestimated their previous activity levels. Using current activity as a proxy for recalled levels did not improve the moderate agreement with activity as originally reported in 1968. The average magnitude of the recall bias (recalled minus original values) amounted to one half of a unit on the four-point scale and was not significantly associated with age or body mass index. In summary, systematic errors were observed in remote physical activity recalls by elderly women, who generally remembered being more active 32 years earlier than they had originally reported. Physical activity recall questionnaires for the distant past should be used with caution, particularly in older populations. PMID- 14742292 TI - A validation study of patient interview data and pharmacy records for antihypertensive, statin, and antidepressant medication use among older women. AB - A validation study evaluated the accuracy of self-reported use of commonly used medications among older women. Within a case-control study of breast cancer, drug information was ascertained by interview. Pharmacy records from 1990 to 1999 were obtained from a Washington State health maintenance organization (66% of subjects) and retail pharmacies (34% of subjects) on a sample of subjects (212 cases, 191 controls) and used as the "gold standard." Recall accuracy was assessed for 6-month, 2-year, and 8-year time windows. Sensitivity of antihypertensive use was 92% (95% confidence interval (CI): 85, 96) for cases and controls in the 6-month period and slightly lower for the 2-year (90% (95% CI: 82, 94) and 87% (95% CI: 78, 92)) and 8-year (80% (95% CI: 69, 88) and 79% (95% CI: 68, 88)) periods. For statins, sensitivity was 83% (95% CI: 64, 93) for cases and 93% (95% CI: 69, 99) for controls in the 6-month period, 75% (95% CI: 55, 88) and 86% (95% CI: 60, 96) in the 2-year period, and 67% (95% CI: 42, 85) and 75% (95% CI: 41, 93) in the 8-year period. For self-report of antidepressants, sensitivities ranged from 66% (95% CI: 47, 80) in the 6-month period to 44% (95% CI: 30, 60) in the 8-year period. Specificity was high among all drug classes, ranging from 91% to 100%. Recall did not differ by case-control status. Trivial changes in estimates were observed when health maintenance organization records alone were used as the gold standard. Self-reported use of antihypertensives and statins appears to be relatively accurate among older women. PMID- 14742293 TI - A biphasic response to silica: I. Immunostimulation is restricted to the early stage of silicosis in lewis rats. AB - Inhalation of crystalline silica may lead to acute or chronic silicosis. Although chronic silicosis is associated with increased incidence/exacerbation of autoimmune disorders, the immunologic effects of chronic silicosis are not completely understood. In an animal model of chronic silicosis, Lewis rats were exposed to filtered air or silica (1.75 microm average particle size) at an exposure concentration of 6.2 mg/m(3), 6 h/d, 5 d/wk for 6 wk, and observed up to 27 wk after the exposure. Based on silica burden, lung histopathology, and immunologic changes, two distinct stages were identified in the development of chronic silicosis. Stage 1 (4-28 d after exposure) was characterized by silica deposition in various tissues, and augmented antibody and cellular immunity. Although bronchoalveolar lavage contained an increased number of activated macrophages, protein and lactate dehydrogenase levels were comparable to controls. In Stage 2 (>/= 10 wk), silica was localized in epithelioid macrophages, and T cell immunity had returned to normal, but the lavage fluids contained increased protein concentration and lactate dehydrogenase activity. Moreover, lungs from silica-treated animals contained neutrophils and lymphocytes, and exhibited granulomatous changes around the silica-containing epithelioid macrophages. Thus, in the early stages of silicosis, silica activates the immune system; however, the progression of lung granulomas does not depend on a continually activated adaptive immune system. PMID- 14742294 TI - The splicing and fate of ADAM33 transcripts in primary human airways fibroblasts. AB - The ADAM (A Disintegrin and Metalloprotease) family of Zn++-dependent metalloproteases are multidomain proteins involved in diverse cellular activities. Polymorphic variation in ADAM33 is strongly associated with asthma and bronchial hyperresponsiveness. Identification of those isoforms of ADAM33 that are expressed in airways is fundamental to dissecting the role of ADAM33 in asthma. Analysis of primary human airways fibroblasts has shown the presence of a number of alternatively spliced forms of ADAM33, including one encoding a putative secreted variant, and many transcripts lacking the metalloproteinase domain. The relative abundance of these transcripts has been quantified using reverse transcription real-time polymerase chain reaction, in both nuclear and cytoplasmic fractions of RNA. These results demonstrate that a number of splice variants of ADAM33 are transported into the cytoplasm. Ninety percent of ADAM33 mRNA is retained in the nucleus and the subtle differences in the composition of nuclear and cytoplasmic RNA suggest important events in the splicing and selection of ADAM33 transcripts. Western blot analysis confirmed that several protein isoforms of ADAM33 are expressed in primary airways fibroblasts. These findings demonstrate that ADAM33 exists in multiple isoforms, suggesting that it is a complex molecule that plays multiple roles within mesenchymal cells. PMID- 14742295 TI - Curcumin-induced apoptosis in scleroderma lung fibroblasts: role of protein kinase cepsilon. AB - Scleroderma, a disease involving excessive collagen deposition, can be studied using fibroblasts cultured from affected tissues. We find that curcumin, the active component of the spice turmeric, causes apoptosis in scleroderma lung fibroblasts (SLF), but not in normal lung fibroblasts (NLF). This effect is likely to be linked to the fact that although curcumin induces the expression of the phase 2 detoxification enzymes heme oxygenase 1 and glutathione S-transferase P1 (GST P1) in NLF, SLF are deficient in these enzymes, particularly after curcumin treatment. The sensitivity of cells to curcumin-induced apoptosis and the expression of GST P1 (but not heme oxygenase 1) are regulated by the epsilon isoform of protein kinase C (PKCepsilon). SLF, which contain less PKCepsilon and less GST P1 than NLF, become less sensitive to curcumin-induced apoptosis and express higher levels of GST P1 when transfected with wild-type PKCepsilon, but not with dominant-negative PKCepsilon. Conversely, NLF become sensitive to curcumin-induced apoptosis and express lower levels of GST P1 when PKCepsilon expression or function is inhibited. The subcellular distribution of PKCepsilon also differs in NLF and SLF. PKCepsilon is predominantly nuclear or perinuclear in NLF but is associated with stress fibers in SLF. Just as PKCepsilon levels are lower in SLF than in NLF in vitro, PKCepsilon expression is decreased in fibrotic lung tissue in vivo. In summary, our results suggest that a signaling pathway involving PKCepsilon and phase 2 detoxification enzymes provides protection against curcumin-induced apoptosis in NLF and is defective in SLF. These observations suggest that curcumin may have therapeutic value in treating scleroderma, just as it has already been shown to protect rats from lung fibrosis induced by a variety of agents. PMID- 14742296 TI - Inhibition of early airway neutrophilia does not affect development of airway hyperresponsiveness. AB - The effect of modifying early neutrophil-mediated inflammation on the development of airway hyperresponsiveness (AHR) was investigated using an interleukin (IL)-1 receptor antagonist (IL-1Ra), an anti-IL-18 antibody (anti-IL-18) or a p38 mitogen-activated protein kinase (MAPK) inhibitor (M39). Balb/c mice were sensitized to ovalbumin (OVA) and challenged with a single intranasal dose of OVA. Treatment with the IL-1Ra or anti-IL-18 was initiated 20 min before challenge, whereas M39 was administered 4 h before the challenge. Eight hours after challenge, sensitized mice showed significantly higher numbers of neutrophils in bronchoalveolar lavage (BAL) fluid; treatment with IL-1Ra, anti-IL 18, or M39 significantly decreased the influx of neutrophils. At 48 h, none of the treatments affected eosinophil inflammation in BAL fluid and lung tissue, goblet cell hyperplasia, or cytokine levels (IL-4, IL-5, IL-12, IL-13, interferon gamma) in BAL fluid. Anti-IL-18 or IL-1Ra had no effect on the development of AHR, whereas M39-treated mice showed a decrease in methacholine responsiveness. These results demonstrate that early neutrophil influx following allergen challenge is mediated by IL-1, IL-18, and p38 MAPK. However, neutralization of IL 1 and IL-18 did not affect the later development of AHR and eosinophilic airway inflammation. The effects of inhibiting p38 MAPK in decreasing AHR indicate activities independent of its prevention of neutrophil accumulation. PMID- 14742297 TI - Keratinocyte growth factor stimulates alveolar type II cell proliferation through the extracellular signal-regulated kinase and phosphatidylinositol 3-OH kinase pathways. AB - Keratinocyte growth factor (KGF or FGF-7) stimulates alveolar type II cell proliferation, but little is known about the signaling pathways involved. We investigated the role of the ERK (p42/44 mitogen activated protein [MAP] kinase) and phosphatidylinositol 3-OH kinase (PI3 kinase) pathways on alveolar type II cell proliferation and differentiation. Rat type II cells were cultured on tissue culture plastic and Matrigel in the presence or absence of KGF and specific chemical inhibitors PD98059, LY294002, and rapamycin at various concentrations. Proliferation was measured by thymidine incorporation and DNA quantitation, and differentiation was measured by expression of surfactant protein A and alkaline phosphatase. We demonstrate that KGF activates distal effectors of the PI3 kinase pathway, PKB/Akt, and p70S6 kinase, as well as p42/44 MAP kinase proteins. Inhibition of these pathways with PD98059, LY294002, or rapamycin inhibited type II cell proliferation but had no significant effect on differentiation. KGF did not activate the c-Jun kinase or p38 MAP kinase pathways. We conclude that the p42/44 MAP kinase and PI3 kinase pathways are important in regulating alveolar type II cell proliferation in response to KGF. PMID- 14742298 TI - Sphingosine kinase mediates activation of extracellular signal-related kinase and Akt by respiratory syncytial virus. AB - Respiratory syncytial virus (RSV) preferentially infects lung epithelial cells. Infected cells remain viable well into the infection. This prolonged survival results from RSV-induced activation of pro-survival pathways, including Akt and extracellular signal-related kinase (ERK). Sphingosine 1-phosphate (S1P) is a sphingolipid metabolite with demonstrated links to cell survival. It is enzymatically generated by sequential activation of ceramidase (generation of sphingosine) and sphingosine kinase (generation of S1P). In these studies, we found that RSV stimulated neutral ceramidase and sphingosine kinase activities in lung epithelial cells. The combined effect of activation of these two enzymes would decrease proapoptotic ceramide and increase antiapoptotic S1P. S1P activated Akt and ERK within minutes, and inhibition of sphingosine kinase blocked RSV-induced ERK and Akt activation, leading to accelerated cell death after viral infection. RSV infection does eventually kill infected cells but activation of cell survival pathways significantly delays cell death. The studies are the first evidence linking sphingolipid metabolites to cell survival mechanisms in the context of a viral infection. PMID- 14742299 TI - Obesity and asthma: directions for research. PMID- 14742300 TI - Lung function, bronchial responsiveness, and asthma in a community cohort of 6 year-old children. AB - KEYWORDS: Children as young as 6 years old can perform spirometry, yet the relationship between current asthma, lung function, and bronchial responsiveness has not been described at this age; 2,537 children from a community-based birth cohort were assessed at 6 years of age, with history (n = 2,141), physical examination (n = 1,995), standard spirometry (n = 1,735), and a random sample (n = 711) offered methacholine challenge. Males had greater values of FVC and FEV(1) but not of mean forced expiratory flow during the middle half of the FVC or FEV(1)/FVC than females. The greatest influences on lung function at 6 years were height, sex, birth weight, and wheezing in the first year of life. Children with current asthma had small but significant deficits in lung function and were more sensitive to methacholine. The optimal cutpoint for determining heightened bronchial responsiveness was found to be a 15% fall in FEV(1) at a dose of 1.8 mg/ml. A negative test could be useful in excluding a diagnosis of asthma (negative predictive value of 92%). Lung function testing, including methacholine challenge, is feasible in 5- to 7-year-old children and has the potential to contribute to the clinical management of children with asthma. PMID- 14742301 TI - Treatment and outcome analysis of 205 patients with multidrug-resistant tuberculosis. AB - Multidrug-resistant tuberculosis, a disease caused by Mycobacterium tuberculosis strains that are resistant at least to rifampin and isoniazid, entails extended treatment, expensive and toxic regimens, and higher rates of treatment failure and death. We retrospectively analyzed the outcomes in 205 patients treated at our center for multidrug-resistant tuberculosis, with strains resistant to a median of six drugs, and compared the results with those of our previous series. Logistic regression and survival analysis were used to evaluate short- and long term outcomes, respectively. Initial favorable response, defined as at least three consecutive negative sputum cultures over a period of at least 3 months, was 85% compared with 65% in the prior cohort. The current cohort had greater long-term success rates, 75% versus 56%, and lower tuberculosis death rates, 12% versus 22%, than the earlier one. Surgical resection and fluoroquinolone therapy were associated with improved microbiological and clinical outcomes in the 205 patients studied after adjusting for other variables. The improvement was statistically significant for surgery and among older patients for fluoroquinolone therapy. PMID- 14742302 TI - Latency and persistence of respiratory syncytial virus despite T cell immunity. AB - Respiratory syncytial virus (RSV) causes bronchiolitis in infants, which is associated with recurrent wheezing in later childhood. There is mounting evidence that the virus becomes latent or persists in vivo, but little is known about the mechanisms of its latency, persistence, and immune evasion. We therefore infected BALB/c mice intranasally with human RSV, analyzed sequential tissue samples by direct culture and polymerase chain reaction for viral and messenger RNA, and monitored antiviral immune responses. Virus could not be detected in bronchoalveolar lavage samples beyond Day 14, but viral genomic and messenger RNA was present in lung homogenates for 100 days or more; combined depletion of CD4 and CD8 T cells allowed infective virus to be recovered. Neutralizing antibody and memory cytotoxic T cell responses were intact in mice with latent infections, and latent viral genome contained an authentic nonmutated M2 82-91 K(d) cytotoxic T lymphocyte epitope. A mutation of this epitope, detected in one clone, did not assist evasion. We suggest that RSV latency depends on persistence in privileged sites rather than on viral mutation. PMID- 14742303 TI - Pulmonary arteriovenous malformations in patients with hereditary hemorrhagic telangiectasia. AB - Pulmonary arteriovenous malformations (PAVMs) associated with hereditary hemorrhagic telangiectasia may cause severe cerebral complications that may be prevented by embolization therapy. We retrospectively compared the diagnostic value of noninvasive tests for the screening of treatable (amenable to embolization) PAVMs in a series of 105 patients, using chest computerized tomography (CT) and/or pulmonary angiography as a "gold standard." Patients had assessment of dyspnea, chest radiograph, alveolar-arterial PO2 gradient under 100% oxygen (AaPO2), contrast echocardiography, and radionuclide perfusion lung scanning. Contrast echocardiography in the supine position was the most sensitive test (93%). The sensitivity of self-reported dyspnea (59%), chest radiograph alone (70%), measurement of AaPO2 by the 100% oxygen method (62%), or radionuclide lung scanning (71%), was not suitable for efficient screening. A 100% sensitivity and negative predictive value could be obtained when combining anteroposterior chest radiograph and contrast echocardiography. Our data support a screening algorithm based on the combined use of contrast echocardiography and anteroposterior chest radiograph, followed by chest CT if either test is positive. An alternative is to screen directly by chest CT. However, this algorithm may obviate the need for chest CT in patients without PAVM, who represent a majority of patients with hereditary hemorrhagic telangiectasia. PMID- 14742304 TI - Airway smooth muscle: the appendix of the lung. PMID- 14742305 TI - The effect of pranlukast on allergen-induced bone marrow eosinophilopoiesis in subjects with asthma. AB - We investigated the mechanisms by which leukotriene receptor antagonists decrease airway eosinophil number. In a randomized, double-blind crossover study, we examined the effects of 2 weeks of treatment with pranlukast 300 mg twice a day or placebo on allergen-induced changes in airway eosinophil number and bone marrow eosinophil progenitors in 15 subjects with mild asthma. Pranlukast treatment for 2 weeks decreased mean sputum eosinophil count from 0.15 x 10(6)/g (5.3% of cells) before treatment to 0.02 x 10(6)/g (0.7% of cells) after treatment (p < 0.05), whereas placebo did not. Pranlukast also decreased the eosinophil count (5.6% at 7 hours and 7.5% at 24 hours) (p < 0.05) after allergen inhalation compared with placebo (13.8% at 7 hours and 15.3% at 24 hours). There was a similar trend for sputum cells immunostaining for EG2, eotaxin, interleukin 5, and regulated upon activation, normal T cell expressed and secreted. Pranlukast also significantly attenuated the allergen-induced increase in the number of bone marrow eosinophil/basophil cfu (mean 0.3) at 24 hours compared with placebo (mean 6.2). The proportion of CD34(+) cells expressing the eotaxin receptor CC chemokine receptor 3, 24 hours after allergen inhalation, was also reduced by pranlukast. We conclude that, the cysteinyl leukotriene receptor antagonist, pranlukast, attenuates allergen-induced increase in airway eosinophils by decreasing bone marrow eosinophilopoiesis and airway chemotactic and eosinophilopoietic cytokines. PMID- 14742306 TI - Activation of 5-HT1A receptors in raphe pallidus inhibits leptin-evoked increases in brown adipose tissue thermogenesis. AB - To elucidate the central neural pathways contributing to the thermogenic component of the autonomic response to intravenous administration of leptin, experiments were conducted in urethane-chloralose-anesthetized, ventilated rats to address 1) the role of neurons in the rostral ventromedial medulla, including raphe pallidus (RPa), in the leptin-evoked stimulation of brown adipose tissue (BAT) sympathetic nerve activity (SNA); and 2) the potential thermolytic effect of 5-hydroxytryptamine(1A) (5-HT(1A)) receptors on RPa neurons that influence BAT thermogenesis. Leptin (1 mg/kg) administration increased BAT SNA by 1,219% of control, BAT temperature by 2.8 degrees C, expired CO(2) by 1.8%, heart rate by 90 beats/min, and mean arterial pressure by 12 mmHg. Microinjection of the 5 HT(1A) receptor agonist 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT) into RPa resulted in a prompt and sustained reversal of the leptin-evoked stimulation of BAT SNA, BAT thermogenesis, and heart rate, with these variables returning to their pre-leptin control levels. Subsequent microinjection of the selective 5 HT(1A) receptor antagonist WAY-100635 into RPa reversed the BAT thermolytic effects of 8-OH-DPAT, returning BAT SNA and BAT temperature to the elevated levels after leptin. In conclusion, activation of neurons in RPa, possibly BAT sympathetic premotor neurons, is essential for the increases in BAT SNA, BAT thermogenesis, and heart rate stimulated by intravenous administration of leptin. Neurons in RPa express 5-HT(1A) receptors whose activation leads to reversal of the BAT thermogenic and the cardiovascular responses to intravenous leptin, possibly through hyperpolarization of local sympathetic premotor neurons. These results contribute to our understanding of central neural substrates for the augmented energy expenditure stimulated by leptin. PMID- 14742307 TI - Effects of vascular endothelial growth factor on isolated fetal alveolar type II cells. AB - Previous investigations gained from in vivo or lung explant studies suggested that VEGF is an autocrine proliferation and maturation factor for developing alveolar type II cells. The objective of this work was to determine whether VEGF exerted its growth and maturation effects directly on isolated type II cells. These were isolated from 19-day fetal rat lung and cultured in defined medium. The presence of VEGF receptor-2 was assessed in cultured cells at the pre- and posttranslational levels. Recombinant VEGF(165), formerly found to be active on lung explants, failed to enhance type II cell proliferation estimated by thymidine and 5-bromo-2'-deoxy-uridine incorporation. It increased choline incorporation in saturated phosphatidylcholine by 27% but did not increase phospholipid surfactant pool size. VEGF (100 ng/ml) left unchanged the transcript level of surfactant proteins (SP)-A, SP-C, and SP-D but increased SP-B transcripts to four times the control steady-state level. VEGF slightly retarded, but did not prevent, the in vitro transdifferentiation of type II into type I cells, as assessed by immunolabeling of the type I cell marker T1alpha. We conclude that, with the exception of SP-B expression, which appears to be controlled directly, the previously observed effects of this VEGF isoform on type II cells are likely to be exerted indirectly through reciprocal paracrine interactions involving other lung cell types. PMID- 14742308 TI - Proteomic analysis of pulmonary edema fluid and plasma in patients with acute lung injury. AB - Proteomics is the large-scale analysis of protein profiles. This approach has not yet been reported in the study of acute lung injury (ALI). This study details protein profiles in plasma and pulmonary edema fluid (EF) from 16 ALI patients and plasma and bronchoalveolar lavage fluid (BALF) from 12 normal subjects. More than 300 distinct protein spots were evident in the EF and BALF of both normal subjects and ALI patients. Of these, 158 were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. In the plasma and EF protein profile of ALI patients, there were multiple qualitative changes. For instance, in all normal subjects, but in only one of the ALI patients, seven distinct surfactant protein A isoforms were evident. Nearly all ALI patients also had protein spots that indicate truncation or other posttranslational modifications. Several of these novel changes could serve as new biomarkers of lung injury. PMID- 14742309 TI - Letter to the Glyco-Forum: The Karl Meyer Award for glycoconjugate research. PMID- 14742311 TI - Aflatoxin B1 formamidopyrimidine adducts are preferentially repaired by the nucleotide excision repair pathway in vivo. AB - Aflatoxin B(1) (AFB(1)), the most potent member of the aflatoxin family of hepatocarcinogens, upon metabolic activation reacts with DNA and forms a population of covalent adducts. The most prevalent adduct, 8,9-dihydro-8-(N(7) guanyl-)-9-hydroxyaflatoxin (AFB(1)-N(7)-dG), as well as the AFB(1) formamidopyrimidine adduct (AFB(1)-FAPY), resulting from imidazole ring opening of the major adduct, are thought to be responsible for mutations caused by AFB(1). The AFB(1)-N(7)-dG lesions are rapidly removed in Escherichia coli and mammals, whereas the AFB(1)-FAPY lesions persist in mammalian cells, which along with the higher stability of this lesion suggests that AFB(1)-FAPY might significantly contribute to the observed toxicity and carcinogenicity of AFB(1) in higher organisms. Other workers have shown in vitro evidence that AFB(1)-FAPY lesions are substrates for both nucleotide excision repair (NER) and base excision repair (BER). The present study, done in vivo, utilized a modified host cell reactivation assay and showed that AFB(1)-FAPY lesions are preferentially repaired in E.coli by NER. Comparisons of repair in wild-type, NER-deficient (uvrA), BER-deficient (mutM) and NER/BER double mutant E.coli strains transformed with plasmids enriched for AFB(1)-N(7)-dG or AFB(1)-FAPY lesions indicate that both lesions are efficiently repaired by NER-proficient cells (both wild-type and BER-deficient strains). We have found that the level of activity of the reporter gene is significantly affected by the presence of either lesion in NER-deficient strains due to the lack of repair. This effect is similar in NER-deficient and NER/BER-deficient strains indicating that BER (specifically in the strains we investigated) does not contribute significantly to the repair of these lesions in vivo. Consistent with this finding, in vitro analysis of AFB(1)-FAPY adduct excision by purified MutM and its functional analog human 8-oxoguanine DNA glycosylase using site-specifically modified oligonucleotides indicates that this lesion is a poor substrate for both proteins compared with canonical substrates for these enzymes, such as 7,8-dihydro-8-oxoguanine and methylformamidopyrimidine. PMID- 14742312 TI - Plutonium targets the p16 gene for inactivation by promoter hypermethylation in human lung adenocarcinoma. AB - Lung cancer from radon or (239)plutonium exposure has been linked to alpha particles that damage DNA through large deletions and point mutations. We investigated the involvement of an epigenetic mechanism, gene inactivation by promoter hypermethylation in adenocarcinomas from plutonium-exposed workers at MAYAK, the first Russian nuclear enterprise established to manufacture weapons plutonium. Adenocarcinomas were collected retrospectively from 71 workers and 69 non-worker controls. Lung adenocarcinomas were examined from workers and non worker controls for methylation of the CDKN2A (p16), O(6)-methylguanine-DNA methyltransferase (MGMT), death associated protein kinase (DAP-K), and Ras effector homolog 1 genes (RASSF1A). The prevalence for methylation of the MGMT or DAP-K genes did not differ between workers and controls, while a higher prevalence for methylation of the RASSF1A gene was seen in tumors from controls. In marked contrast, the prevalence for methylation of p16, a key regulator of the cell cycle, was increased significantly (P = 0.03) in tumors from workers compared with non-worker controls. Stratification of plutonium exposure into tertiles also revealed a striking dose response for methylation of the p16 gene (P = 0.008). Workers in the plutonium plant where exposure to internal radiation was highest had a 3.5 times (C.I. 1.5, 8.5; P = 0.001) greater risk for p16 methylation in their tumors than controls. This increased probability for methylation approximated the 4-fold increase in relative risk for adenocarcinoma in this group of workers exposed to plutonium. In addition, a trend (P = 0.08) was seen for an increase in the number of genes methylated (> or =2 genes) with plutonium dose. Here we demonstrate that exposure to plutonium may elevate the risk for adenocarcinoma through specifically targeting the p16 gene for inactivation by promoter methylation. PMID- 14742313 TI - Proanthocyanidins from grape seeds inhibit expression of matrix metalloproteinases in human prostate carcinoma cells, which is associated with the inhibition of activation of MAPK and NF kappa B. AB - Prostate cancer (PCA) is the second most frequently diagnosed and leading cause of cancer-related deaths in men in the USA. The recognition that matrix metalloproteinases (MMPs) facilitate tumor cell invasion and metastasis of PCA has led to the development of MMP inhibitors as cancer therapeutic agents. As part of our efforts to develop newer and effective chemopreventive agents for PCA, we evaluated the effect of proanthocyanidins from grape seeds (GSP) on metastasis-specific MMP-2 and -9 in human prostate carcinoma DU145 cells by employing western blot and gelatinolytic zymography. Treatment of GSP dose dependently inhibited cell proliferation (15-100% by 5-80 microg/ml of GSP), viability (30-80% by 20-80 microg/ml of GSP) and fibroblast conditioned medium (FCM)-induced expression of MMP-2 and -9 in DU145 cells. Since the signaling cascade of mitogen-activated protein kinases (MAPK) have been shown to regulate the expression of MMPs in tumor cells, we found that the treatment of DU145 cells with GSP (20-80 microg/ml) resulted in marked inhibition of FCM-induced phosphorylation of extracellular signal regulated kinase (ERK)1/2 and p38 but had little effect on c-Jun N-terminal kinase under similar experimental conditions. GSP treatment (20-80 microg/ml) to DU145 cells also dose-dependently inhibited FCM-induced activation of NF kappa B concomitantly with inhibition of MMP-2 and 9 expression in the same system. Additionally, the treatment of inhibitors of MEK (PD98059) and p38 (SB203580) to DU145 cells resulted in the reduction of FCM induced phosphorylation of ERK1/2 and p38 concomitantly marked reduction in MMP-2 and -9 expressions. In further studies, treatment of androgen-sensitive LNCaP cells with a synthetic androgen R1881, resulted in an increase of MMP-2 and -9, which were completely abrogated in the presence of GSP (20-60 microg/ml). These data suggest that inhibition of metastasis-specific MMPs in tumor cells by GSP is associated with the inhibition of activation of MAPK and NF kappa B pathways, and thus provides the molecular basis for the development of GSP as a novel chemopreventive agent for both androgen-sensitive and -insensitive prostate cancer therapies. PMID- 14742314 TI - An acyclic retinoid, NIK-333, inhibits N-diethylnitrosamine-induced rat hepatocarcinogenesis through suppression of TGF-alpha expression and cell proliferation. AB - The present study was designed to determine the effects of NIK-333, a synthetic acyclic retinoid, on N-diethylnitrosamine (DEN)-induced hepatocarcinogenesis in male F344 rats. Animals were given DEN dissolved in drinking water at a concentration of 40 p.p.m. for 5 weeks and then provided with drinking water free of DEN for 15 weeks to induce hepatocellular neoplasms. NIK-333 was administered orally (once a day) to rats at doses of 10, 40 and 80 mg/kg body wt for 14 weeks, starting 1 week after the completion of administration of DEN. At 20 weeks after the start of DEN administration, histopathological evaluation was carried out on all animals. The effects of NIK-333 on the cell proliferation activity of non tumorous areas and liver tumor cells and the immunohistochemical expression of transforming growth factor-alpha (TGF-alpha) were also evaluated. NIK-333 at 40 and 80 mg/kg body wt significantly inhibited hepatocarcinogenesis (P < 0.05). In addition, NIK-333 at the same doses decreased DEN-induced overexpression of TGF alpha in hepatocellular neoplasms (adenomas and carcinomas) and their surrounding tissue. Furthermore, NIK-333 significantly inhibited cell proliferation activity in the lesions and in non-tumorous areas (P < 0.01). Our results suggest that NIK 333 inhibits DEN-induced hepatocarcinogenesis through suppression of TGF-alpha expression and cell proliferation. PMID- 14742315 TI - Recombination at chromosomal sequences involved in leukaemogenic rearrangements is differentially regulated by p53. AB - Chromosomal translocations and retroviral integration events at breakpoint cluster regions (bcrs) have been associated with leukaemias. To directly compare the effect of different cis-regulatory sequences on recombination, we adapted our SV40 based model system to the analysis of correspondingly selected bcrs from the TAL1, LMO2, retinoic acid receptor alpha (RARalpha) and MLL genes. We show that a 399 bp fragment from the MLL bcr is sufficient to cause a 3-4-fold stimulation of spontaneously occurring DNA exchange and to respond to etoposide by up to 10-fold further elevated frequencies, i.e. to mimic the fragility of the 8.3 kb bcr during chemotherapy. To analyse the regulatory role of p53 in recombination involving leukaemia-related sequences, we stably expressed wtp53 and a transactivation negative mutant. Consistent with the proposed role of p53 as a suppressor of error-prone recombination, both p53 proteins down-regulated recombination with most of the sequences tested, even with the MLL bcr after etoposide treatment. Surprisingly, however, p53 stimulated recombination, in constructs carrying the RARalpha bcr fragment. This is the first study, which provides evidence for a stimulatory role of p53 in homologous recombination. Our data further indicate that inhibition of topoisomerase I can mimic the effects of p53 on stimulating recombination on the RARalpha bcr. Thus, these data also firstly describe a biological role of the biochemical interactions between p53 and topoisomerase I that may have implications for a gain-of-function phenotype of certain p53 mutants in genetic destabilization. PMID- 14742316 TI - Carcinogenic properties of proteins with pro-inflammatory activity from Streptococcus infantarius (formerly S.bovis). AB - Several studies reported linkage between bacterial infections and carcinogenesis. Streptococcus bovis was traditionally considered as a lower grade pathogen frequently involved in bacteremia and endocarditis. This bacterium became important in human health as it was shown that 25-80% of patients who presented a S.bovis bacteremia had also a colorectal tumor. Moreover, in previous experiments, we demonstrated that S.bovis or S.bovis wall extracted antigens (WEA) were able to promote carcinogenesis in rats. The aim of the present study was: (i) to identify the S.bovis proteins responsible for in vitro pro inflammatory properties; (ii) to purify them; (iii) to examine their ability to stimulate in vitro IL-8 and COX-2 expression by human colon cancer cells; and (iv) to assess in vivo their pro-carcinogenic potential in a rat model of colon carcinogenesis. The purified S300 fraction, as determined by proteomic analysis, contained 72 protein spots in two-dimensional gel electrophoresis representing 12 different proteins able to trigger human epithelial colonic Caco-2 cells and rat colonic mucosa to release CXC chemokines (human IL-8 or rat CINC/GRO) and prostaglandins E2, correlated with an in vitro over-expression of COX-2. Moreover, these proteins were highly effective in the promotion of pre-neoplastic lesions in azoxymethane-treated rats. In the presence of these proteins, Caco-2 cells exhibited enhanced phosphorylation of the three classes of MAP kinases. Our results show a relationship between the pro-inflammatory potential of S.bovis proteins and their pro-carcinogenic properties, confirming the linkage between inflammation and colon carcinogenesis. These data support the hypothesis that colonic bacteria can contribute to cancer development particularly in chronic infection/inflammation diseases where bacterial components may interfere with cell function. PMID- 14742317 TI - Immunohistochemical detection of 1,N6-ethenodeoxyadenosine in nuclei of human liver affected by diseases predisposing to hepato-carcinogenesis. AB - Increased oxidative stress and lipid peroxidation (LPO) are implicated in multistage carcinogenesis. Recent studies have shown that LPO-derived reactive hydroxyalkenals can form promutagenic exocyclic etheno-DNA adducts in vivo. Such DNA damage was found to be increased in the liver of patients with metal storage diseases and in colon adenomas of familial adenomatous polyposis patients. We now have investigated the levels of 1,N(6)-ethenodeoxyadenosine (epsilon dA) in human liver samples obtained from a group of patients diagnosed with hepatitis, fatty liver, fibrosis and cirrhosis, primary hemochromatosis and Wilson's disease. Using an immunohistochemical method, the relative mean pixel intensity of randomly selected nuclei was measured by imaging software; positively stained cell nuclei (arbitrary mean pixel intensity > or =0.5) were counted. Prevalence of epsilon dA (%) was calculated from the ratio of a number of positively stained cell nuclei over a total number of cells counted. When compared with normal livers (3.1%), the percent prevalence (means) was significantly higher in specimens of alcoholic fatty liver (15%) and fibrosis patients (50%) but not in samples with hepatitis (induced by various factors) (6.2%). The percent prevalence in alcohol fibrosis was as high as in the liver from Wilson's disease (50.7%) and hemochromatosis (33%) patients. This is the first demonstration of increased epsilon dA in human liver diseases due to alcohol abuse. We conclude that excessive hepatic DNA damage, as assessed by miscoding etheno-DNA adduct in the nuclei of liver biopsies, is probably caused by alcohol-induced oxidative stress and LPO. In cancer-prone liver diseases (fatty liver, cirrhosis/fibrosis) such damage may act as a driving force towards malignancy. PMID- 14742318 TI - Diet and alcohol consumption in relation to p53 mutations in breast tumors. AB - There is evidence linking alcohol consumption to p53 mutations in tumors, considerable evidence linking alcohol consumption with risk of breast cancer and some evidence that alcohol and folate consumption interact to affect risk. Further, while there is some indication that oxidation may play a role in breast cancer etiology, there has been little examination of an association of oxidative stress with p53 mutations. We examined several dietary components related to one carbon metabolism and antioxidants to determine if these factors were related to the prevalence of p53 mutations in breast tumors. We conducted a case-control study of primary, histologically confirmed breast cancer in western New York. Controls <65 were selected from drivers license lists; those > or =65 were selected from Health Care Finance Administration lists. p53 mutations in archived tumor blocks were identified in exons 2-11 and flanking intron sequences. Usual dietary intake was assessed by interview regarding intake in the previous 2 years; alcohol consumption was queried for 2, 10 and 20 years in the past. Our data were consistent with increased likelihood of tumors with p53 mutations for premenopausal breast cancer with increased alcohol intake 10 or 20 years previous; for intake of 16 or more drinks per month in the period 20 years before the interview compared with non-drinkers, the OR was 5.25, 95% CI 1.48-18.58. For postmenopausal women, there was increased likelihood of tumors with p53 mutations among women with higher folate. Antioxidant nutrients were not differentially related to p53 mutations. These results indicate that there may be heterogeneity in breast tumors, as indicated by differences in associations for those with or without p53 mutations, and that causal pathways for these nutrients may vary for pre- and postmenopausal women. For premenopausal women, alcohol consumption in the past was associated with p53 mutations. PMID- 14742319 TI - Id-1-induced Raf/MEK pathway activation is essential for its protective role against taxol-induced apoptosis in nasopharyngeal carcinoma cells. AB - Increasingly, evidence supports the function of the helix-loop-helix protein Id-1 (inhibitor of differentiation/DNA binding-1) as an oncogene. Over-expression of Id-1 is not only observed in many types of human cancer but its expression levels have been correlated with cancer progression. However, little is known about the molecular mechanisms responsible for the function of Id-1. Recently, we and others reported that Id-1-induced cell proliferation was mediated through a Raf/MEK signalling pathway. In this study, we investigated if ectopic Id-1 expression in nasopharyngeal carcinoma cells had any protective effect on taxol induced death, which is also regulated through Raf/MEK pathway. Using four stable Id-1 transfectant clones, we found that exogenous Id-1 expression led to phosphorylation of Raf-1 and MEK1/2 kinases, which was associated with resistance to taxol. Treatment of the Id-1 expressing cells with a MEK inhibitor, PD098059, resulted in an increased taxol-induced apoptosis rate in Id-1 transfectants compared with the vector control. In addition, the fact that the taxol-induced apoptosis rate, down-regulation of Bcl-2 and up-regulation of Bax were suppressed by PD098059 treatment in Id-1 expressing cells indicates that the Id-1 induced cellular protection against apoptosis is mediated through Raf/MEK signalling pathways. Our results suggest that Id-1 may be an upstream regulator of the Raf/MEK signalling pathway, which plays an essential role in protection against taxol-induced apoptosis. Our evidence also indicates a novel treatment strategy to increase anticancer drug-induced apoptosis through inactivation of the Id-1 protein. PMID- 14742320 TI - Pancreatic cancer cells express 25-hydroxyvitamin D-1 alpha-hydroxylase and their proliferation is inhibited by the prohormone 25-hydroxyvitamin D3. AB - The steroid hormone 1,25-dihydroxyvitamin D(3), [1,25(OH)(2)D(3), calcitriol], the active metabolite of vitamin D, exerts pleiotropic antitumor effects against several malignancies. However, the clinical use of this hormone is limited by hypercalcemia. 25-Hydroxyvitamin D(3), the prohormone of 1,25(OH)(2)D(3), is hydroxylated to the active hormone by the enzyme 25-hydroxyvitamin-1-alpha hydroxylase [1 alpha(OH)ase]. 1 alpha(OH)ase is found primarily in the kidney, but also is expressed in the prostate, colon and other tissues. Using immunohistochemistry, we report that 1 alpha(OH)ase is highly expressed in both normal and malignant pancreatic tissue. Expression of this enzyme and enzymatic activity was also detected in four pancreatic tumor cell lines. 25(OH)D(3) inhibited the growth of three of four pancreatic cell lines in a manner that correlated with the level of induction of the cyclin-dependent kinase inhibitors p21 and p27 and with the induction of cell cycle arrest at the G(1)/S checkpoint. The growth of a cell line stably transfected with a mutant Ki-ras allele and of a second cell line with an endogenous Ki-ras activating mutation was also inhibited by 25(OH)D(3), indicating that activating Ki-Ras mutations, which occur in almost 90% of pancreatic adenocarcinomas, do not interfere with the growth-inhibitory effects of 25(OH)D(3). The expression of 1 alpha(OH)ase in normal and malignant pancreatic tissue and the antiproliferative effects of the prohormone in these cells, suggest that 25(OH)D(3) may offer possible therapeutic and chemopreventive options for pancreatic cancer. PMID- 14742321 TI - UV radiation-induced XPC translocation within chromatin is mediated by damaged DNA binding protein, DDB2. AB - The tumor suppressor p53 protein has been established as an important factor in modulating the efficiency of global genomic repair. Our recent repair studies in human cells reported that p53 regulates the recruitment of XPC and TFIIH proteins to specific DNA damage sites. Here, we have examined the influence of p53 and damaged-DNA binding complex (DDB2) proteins on the distribution of XPC within damaged chromatin in vivo and the recruitment of XPC to DNA damage sites in situ. The results show that UV irradiation causes the translocation of XPC from a loosely bound form into a tight association with chromatin in vivo. The UV radiation-induced redistribution of XPC was equally compromised in p53-deficient, as well as DDB2-deficient, human cells. Similarly, rapid recruitment of XPC to DNA damage in situ was also impaired in both cell lines. Ectopic expression of DDB2 in p53-deficient cells overcame the requirement of p53 function for UV induced translocation of XPC in vivo. Restoration of DDB2 function also enhanced the recruitment of XPC to DNA damage sites in situ and increased the global repair of cyclobutane pyrimidine dimer from the genome. These results indicate that DDB2 is a key downstream factor of p53 for regulating the movement of XPC to DNA damage in irradiated cells. PMID- 14742322 TI - Urinary N2-(2'-deoxyguanosin-8-yl)PhIP as a biomarker for PhIP exposure. AB - The food-derived, heterocyclic aromatic amine 2-amino-1-methyl-6 phenylimidazo[4,5-b]pyridine (PhIP) is genotoxic and is carcinogenic in experimental animals. Studies on the role of PhIP in human diet-related cancer would be aided considerably by the availability of a readily applicable biomarker of the internal dose of the ultimate genotoxic species. PhIP has been shown to adduct primarily at C-8 of deoxyguanosine in DNA and so the DNA repair product N(2)-(2'-deoxyguanosin-8-yl)PhIP is a potential biomarker of DNA adduction and repair after exposure to PhIP. An assay for N(2)-(2'-deoxyguanosin-8-yl)PhIP in urine has been developed based on liquid chromatography mass spectrometry, using a deuterated analogue of the nucleoside as an internal standard and an antibody mediated extraction procedure. Polyclonal antibodies were raised against the PhIP nucleotide, PhIP-nucleoside and PhIP-guanine base adducts conjugated to keyhole limpet haemocyanin. Following their evaluation, the immobilized PhIP nucleotide antibody was used for the extraction of N(2)-(2'-deoxyguanosin-8-yl)PhIP from urine. The limit of detection of the assay was 125 pg and the limit of quantification 200 pg for a 50 ml human urine sample. Following oral administration of PhIP (20 mg/kg body wt/day) to rats for 6 days, N(2)-(2' deoxyguanosin-8-yl) PhIP was readily detected in the urine, reaching steady state over 3 days. This is the first direct demonstration of the urinary elimination of this adduct following exposure to parent amine. The half-life of the adduct with DNA was estimated to be approximately 20 h. The total amount of PhIP recovered in the urine as adduct was <0.5 x 10(-3)% of the dose administered. Levels of the PhIP adduct in urine collections from human subjects ingesting the amine (4.9 micro g) in cooked meat were below the limits of detection, indicating that humans are exposed to a bioactive dose of <3 x 10(-4) of that associated with a non-carcinogenic level in rats. PMID- 14742323 TI - Activation of beta-catenin provides proliferative and invasive advantages in c myc/TGF-alpha hepatocarcinogenesis promoted by phenobarbital. AB - Previously, we have found that phenobarbital (PB) enhanced cell survival and facilitated tumor growth in our c-myc/transforming growth factor (TGF)-alpha transgenic mouse model of liver cancer. Given that PB selectively promoted initiated cells harboring beta-catenin mutations during chemically induced hepatocarcinogenesis and that Wnt/beta-catenin signaling is involved in both anti apoptotic and proliferative processes, we now have extended our analysis to investigate whether promotion by PB affects the occurrence of beta-catenin mutations in c-myc/TGF-alpha-driven tumors. The frequency of beta-catenin activation as judged by somatic mutations and/or nuclear localization was significantly increased in hepatocellular carcinomas (HCCs) from c-myc/TGF-alpha mice treated with PB (15/28; 53.6%) as compared with that in control HCCs (2/28; 7.1%). Furthermore, an intact beta-catenin locus was detected in all neoplasms following PB treatment, whereas 57.1% (16/28) of malignant tumors from c-myc/TGF alpha untreated mice displayed loss of heterozygosity at the beta-catenin locus. Strikingly, in the majority of PB-treated HCCs beta-catenin nuclear localization was limited to small cells with high nuclear/cytoplasmic ratio forming an invasion front (NAinv). beta-Catenin NAinv cells showed cytoplasmic redistribution of E-cadherin associated with intense mucin 1 and matrilysin immunostaining, suggesting their invasive phenotype. All beta-catenin-positive HCCs displayed increased proliferation and tumor size, but no difference in the apoptotic rate when compared with beta-catenin negative tumors. These findings show that PB treatment positively selects for a cell population displaying activation of beta-catenin in c-myc/TGF-alpha HCCs. beta-Catenin activation confers additional growth and invasive advantages in a model of liver cancer already accelerated by synergistic activity of the c-myc and TGF-alpha transgenes. PMID- 14742324 TI - RasGTPase-activating protein is a target of caspases in spontaneous apoptosis of lung carcinoma cells and in response to etoposide. AB - p120 RasGTPase-activating protein (RasGAP), the main regulator of Ras GTPase family members, is cleaved at low caspase activity into an N-terminal fragment that triggers potent anti-apoptotic signals via activation of the Ras/PI-3 kinase/Akt pathway. When caspase activity is increased, RasGAP fragment N is further processed into two fragments that effectively potentiate apoptosis. Expression of RasGAP protein and its cleavage was assessed in human lung cancer cells with different histology and responsiveness to anticancer drug-induced apoptosis. Here we show that therapy-sensitive small lung carcinoma cell (SCLC) lines have lower RasGAP expression levels and higher spontaneous cleavage with formation of fragment N compared to therapy-resistant non-small cell lung carcinoma cell (NSCLC) lines. The first RasGAP cleavage event strongly correlated with the increased level of spontaneous apoptosis in SCLC. However, generation of protective RasGAP fragment N also related to the potency of SCLC to develop secondary therapy-resistance. In response to etoposide (ET), RasGAP fragment N was further cleaved in direct dependence on caspase-3 activity, which was more pronounced in NSCLC cells. Caspase inhibition, while effectively preventing the second cleavage of RasGAP, barely affected the first cleavage of RasGAP into fragment N that was always detectable in NSCLC and SCLC cells. These findings suggest that different levels of RasGAP and fragment N might play a significant role in the biology and different clinical course of both subtypes of lung neoplasms. Furthermore, constitutive formation of RasGAP fragment N can potentially contribute to primary resistance of NSCLC to anticancer therapy by ET but also to secondary therapy-resistance in SCLC. PMID- 14742325 TI - Mice deficient for the gap junction protein Connexin32 exhibit increased radiation-induced tumorigenesis associated with elevated mitogen-activated protein kinase (p44/Erk1, p42/Erk2) activation. AB - Loss of connexin expression/gap junction intercellular communication (GJIC) has been correlated with decreased growth control and increased tumorigenesis. Studies utilizing Connexin32 (Cx32)-deficient knockout mice have demonstrated that loss of Cx32 increases susceptibility to chemically induced liver tumorigenesis. Here, in addition to dramatically increased liver tumorigenesis, we show that tumor induction utilizing X-ray radiation resulted in a statistically significant increase in overall tumor burden in Cx32-deficient mice compared with wild-type mice due to tumorigenesis in several other tissues (lung, adrenal, lymph and small intestine) even when excluding prevalent liver tumors. Irradiated Cx32-deficient mice were particularly sensitive to liver tumorigenesis (46% incidence compared with 18% in wild-type mice, P = 0.007) demonstrating that Cx32 functions as a hepatic tumor suppressor in response to radiation-associated mutation events. Cx32-deficient mice also exhibited increased lung tumorigenesis (bronchioloalveolar) with an increased progression to carcinoma when compared with wild-type mice. Two Cx32-deficient mice developed an uncommon, invasive medullary adrenal tumor type (pheochromocytoma) not observed in irradiated wild type mice. Immunohistochemical analysis revealed increased levels of activated mitogen-activated protein kinase (MAPK) (p44/Erk1, p42/Erk2) in Cx32-deficient mouse liver tumors (P = 0.006), lung tumors (P = 0.056) and adrenal tumors (primary and metastases) compared with wild-type counterparts implicating elevated activation of MAPK-interacting pathways in Cx32-deficient tumorigenesis. Interestingly, lung tumors from Cx32-deficient mice also demonstrated decreased p27Kip1 levels compared with wild-type lung tumors (P = 0.05). This study demonstrates that loss of Cx32/GJIC plays a significant role in radiation-induced tumorigenesis of the liver and importantly that Cx32 may also play a role in tumor suppression and/or tumor progression in other tissue types such as lung and adrenal gland. Additionally, this mouse model suggests that MAPK-related pathways may be preferentially activated or conversely that tumors harboring activated MAPK pathways may selectively progress towards more advanced tumor states in the absence of Cx32-mediated GJIC. PMID- 14742326 TI - The Patient State Index as an indicator of the level of hypnosis under general anaesthesia. AB - BACKGROUND: This retrospective study describes the performance of the Patient State Index (PSI), under standard clinical practice conditions. The PSI is comprised of quantitative features of the EEG (QEEG) that display clear differences between hypnotic states, but consistency across anaesthetic agents within the state. METHODS: The PSI was constructed from a systematic investigation of a database containing QEEG extracted from the analyses of continuous 19 channel EEG recordings obtained in 176 surgical patients. Induction was accomplished with etomidate, propofol, or thiopental. Anaesthesia was maintained by isoflurane, desflurane, or sevoflurane, total i.v. anaesthesia using propofol, or nitrous oxide/narcotics. It was hypothesized that a multivariate algorithm based on such measures of brain state, would vary significantly with changes in hypnotic state. RESULTS: Highly significant differences were found between mean PSI values obtained during the different anaesthetic states selected for study. The relationship between level of awareness and PSI value at different stages of anaesthetic delivery was also evaluated. Regression analysis for prediction of arousal level using PSI was found to be highly significant for the combination of all anaesthetics, and for the individual anaesthetics. CONCLUSIONS: The PSI, based upon derived features of brain electrical activity in the anterior/posterior dimension, significantly co varies with changes in state under general anaesthesia and can significantly predict the level of arousal in varying stages of anaesthetic delivery. PMID- 14742327 TI - High-fidelity patient simulation: validation of performance checklists. AB - BACKGROUND: Standardized scenarios can be used for performance assessments geared to the level of the learner. The purpose of this study was to validate checklists used for the assessments of medical students' performance using high-fidelity patient simulation. METHODS: Our undergraduate committee designed 10 scenarios based on curriculum objectives. Fifteen faculty members with undergraduate educational experience identified items considered appropriate for medical students' performance level and identified items that, if omitted, would negatively affect grades. Items endorsed by less than 20% of faculty were omitted. For remaining items, weighting was calculated according to faculty responses. Students managed at least one scenario during which their performance was videotaped. Two raters independently completed the checklists for three consecutive sessions to determine inter-rater reliability. Validity was determined using Cronbach's alpha with an alpha>or=0.6 and 3' exonuclease activity plays important roles in DNA replication and repair. In this study, the kinetic parameters of mutants at highly conserved aromatic residues, Tyr33, Phe35, Phe79, and Phe278-Phe279, in the vicinity of the catalytic centers of FEN-1 were examined. The substitution of these aromatic residues with alanine led to a large reduction in kcat values, although these mutants retained Km values similar to that of the wild-type enzyme. Notably, the kcat of Y33A and F79A decreased 333-fold and 71-fold, respectively, compared with that of the wild type enzyme. The aromatic residues Tyr33 and Phe79, and the aromatic cluster Phe278-Phe279 mainly contributed to the recognition of the substrates without the 3' projection of the upstream strand (the nick, 5'-recess-end, single-flap, and pseudo-Y substrates) for the both exo- and endo-activities, but played minor roles in recognizing the substrates with the 3' projection (the double flap substrate and the nick substrate with the 3' projection). The replacement of Tyr33, Phe79, and Phe278-Phe279, with non-charged aromatic residues, but not with aliphatic hydrophobic residues, recovered the kcat values almost fully for the substrates without the 3' projection of the upstream strand, suggesting that the aromatic groups of Tyr33, Phe79, and Phe278-Phe279 might be involved in the catalytic reaction, probably via multiple stacking interactions with nucleotide bases. The stacking interactions of Tyr33 and Phe79 might play important roles in fixing the template strand and the downstream strand, respectively, in close proximity to the active center to achieve the productive transient state leading to the hydrolysis. PMID- 14742431 TI - Proteasome inhibition alters neural mitochondrial homeostasis and mitochondria turnover. AB - Inhibition of proteasome activity occurs in normal aging and in a wide variety of neurodegenerative conditions including Alzheimer's disease and Parkinson's disease. Although each of these conditions is also associated with mitochondrial dysfunction potentially mediated by proteasome inhibition, the relationship between proteasome inhibition and the loss of mitochondrial homeostasis in each of these conditions has not been fully elucidated. In this study, we conducted experimentation in order to begin to develop a more complete understanding of the effects proteasome inhibition has on neural mitochondrial homeostasis. Mitochondria within neural SH-SY5Y cells exposed to low level proteasome inhibition possessed similar morphological features and similar rates of electron transport chain activity under basal conditions as compared with untreated neural cultures of equal passage number. Despite such similarities, maximal complex I and complex II activities were dramatically reduced in neural cells subject to proteasome inhibition. Proteasome inhibition also increased mitochondrial reactive oxygen species production, reduced intramitochondrial protein translation, and increased cellular dependence on glycolysis. Finally, whereas proteasome inhibition generated cells that consistently possessed mitochondria located in close proximity to lysosomes with mitochondria present in the cellular debris located within autophagosomes, increased levels of lipofuscin suggest that impairments in mitochondrial turnover may occur following proteasome inhibition. Taken together, these data demonstrate that proteasome inhibition dramatically alters specific aspects of neural mitochondrial homeostasis and alters lysosomal mediated degradation of mitochondria with both of these alterations potentially contributing to aging and age-related disease in the nervous system. PMID- 14742432 TI - Subcellular localization and targeting of N-acetylglucosaminyl phosphatidylinositol de-N-acetylase, the second enzyme in the glycosylphosphatidylinositol biosynthetic pathway. AB - The second step in glycosylphosphatidylinositol biosynthesis is the de-N acetylation of N-acetylglucosaminylphosphatidylinositol (GlcNAc-PI) catalyzed by N-acetylglucosaminylphosphatidylinositol deacetylase (PIG-L). Previous studies of mouse thymoma cells showed that GlcNAc-PI de-N-acetylase activity is localized to the endoplasmic reticulum (ER) but enriched in a mitochondria-associated ER membrane (MAM) domain. Because PIG-L has no readily identifiable ER sorting determinants, we were interested in learning how PIG-L is localized to the ER and possibly enriched in MAM. We used HeLa cells transiently or stably expressing epitope-tagged PIG-L variants or chimeric constructs composed of elements of PIG L fused to Tac antigen, a cell surface protein. We first analyzed the subcellular distribution of PIG-L and Glc-NAc-PI-de-N-acetylase activity and then studied the localization of Tac-PIG-L chimeras to identify sequence elements in PIG-L responsible for its subcellular localization. We show that human PIG-L is a type I membrane protein with a large cytoplasmic domain and that, unlike the result with mouse thymoma cells, both PIG-L and GlcNAc-PI-de-N-acetylase activity are uniformly distributed between ER and MAM in HeLa cells. Analyses of a series of Tac-PIG-L chimeras indicated that PIG-L contains two ER localization signals, an independent retention signal located between residues 60 and 88 of its cytoplasmic domain and another weak signal in the luminal and transmembrane domains that functions autonomously in the presence of membrane proximal residues of the cytoplasmic domain that themselves lack any retention information. We conclude that PIG-L, like a number of other ER membrane proteins, is retained in the ER through a multi-component localization signal rather than a discrete sorting motif. PMID- 14742433 TI - Microscopic evidence that actin-interacting protein 1 actively disassembles actin depolymerizing factor/Cofilin-bound actin filaments. AB - Actin-depolymerizing factor (ADF)/cofilin and gelsolin are the two major factors to enhance actin filament disassembly. Actin-interacting protein 1 (AIP1) enhances fragmentation of ADF/cofilin-bound filaments and caps the barbed ends. However, the mechanism by which AIP1 disassembles ADF/cofilin-bound filaments is not clearly understood. Here, we directly observed the effects of these proteins on filamentous actin by fluorescence microscopy and gained novel insight into the function of ADF/cofilin and AIP1. ADF/cofilin severed filaments and AIP1 strongly enhanced disassembly at nanomolar concentrations. However, gelsolin, gelsolin actin complex, or cytochalasin D did not enhance disassembly by ADF/cofilin, suggesting that the strong activity of AIP1 cannot be explained by simple barbed end capping. Barbed end capping by ADF/cofilin and AIP1 was weak and allowed filament elongation, whereas gelsolin or gelsolin-actin complex strongly capped and inhibited elongation. These results suggest that AIP has an active role in filament severing or depolymerization and that ADF/cofilin and AIP1 are distinct from gelsolin in modulating filament elongation. PMID- 14742434 TI - Gene and protein characterization of the human glutathione S-transferase kappa and evidence for a peroxisomal localization. AB - Kappa class glutathione S-transferase (GST) cDNA sequences have been identified in rat, mouse, and human. In the present study, we determined the structure and chromosomal location of the human GST Kappa 1 (hGSTK1) gene, characterized the protein, and demonstrated its subcellular localization. The human gene spans approximately 5 kb, has 8 exons, and maps onto chromosome 7q34. The 5'-flanking region lacks TATA or CCAAT boxes, but there is an initiator element overlapping the transcription start site. hGSTK1 amino acid sequence showed homology to bacterial 2-hydroxychromene-2-carboxylate isomerase, an enzyme involved in naphthalene degradation pathway. hGSTK1 mRNA was expressed in all of the organs examined. Subcellular fractionation of HepG2 cells showed that the protein was located in peroxisomes and mitochondria and was not detectable in cytoplasm. The peroxisomal localization was confirmed by transfection of HepG2 cells with a plasmid coding a green fluorescent protein fused inframe to the N terminus of hGSTK1. The C terminus of hGSTK1 was essential for localization of the protein to peroxisomes, and the C-terminal sequence Ala-Arg-Leu represents a peroxisome targeting signal. This is the first time that a human GST has been found in peroxisomes, suggesting a new function for this family of enzymes. PMID- 14742435 TI - Prostaglandin E2 synergistically enhances receptor tyrosine kinase-dependent signaling system in colon cancer cells. AB - Cyclooxygenase (COX)-generated prostaglandin E(2) (PGE(2)) plays critical roles in colorectal carcinogenesis. Recently, we have shown that PGE(2) and transforming growth factor-alpha synergistically induces the expression of amphiregulin (AR) in colon cancer cells (Shao, J., Evers, B. M., and Sheng, H. (2003) Cancer Res. 63, 5218-5223). In this study, we demonstrated synergistic actions of PGE(2) and the receptor tyrosine kinase signaling system in AR expression and in tumorigenic potential of colon cancer cells. Activation of the Ras/Raf/MAPK pathway induced AR transcription in colon cancer LS-174 cells that was enhanced by PGE(2) in a synergistic fashion. The cAMP-responsive element within the AR promoter was required for the synergistic activation of AR transcription. An Sp1 element was responsible for the basal transcription of AR and significantly enhanced the synergy between PGE(2) and the epidermal growth factor receptor (EGFR) signaling system. Furthermore, activation of both PGE(2) and EGFR signaling pathways synergistically promoted the growth and migration of colon cancer cells. Our results suggest that COX-2/PGE(2) may exert pro-oncogenic effects through synergistic induction of receptor tyrosine kinase-dependent signaling pathway, thus, provide a novel mechanism for the combinatorial treatment of colonic neoplasms targeting both COX-2/PGE(2) and the EGFR system that has demonstrated remarkable advantages. PMID- 14742436 TI - Transcription termination by phage HK022 Nun is facilitated by COOH-terminal lysine residues. AB - The 109-amino acid Nun protein of prophage HK022 excludes superinfecting bacteriophage lambda by blocking transcription elongation on the lambda chromosome. Multiple interactions between Nun and the transcription elongation complex are involved in this reaction. The Nun NH(2)-terminal arginine-rich motif binds BOXB sequence in nascent lambda transcripts, whereas the COOH terminus binds RNA polymerase and contacts DNA template. Nun Trp(108) is required for interaction with DNA and transcription arrest. We analyzed the role of the adjacent Lys(106) and Lys(107) residues in the Nun reaction. Substitution of the lysine residues with arginine (K106R/K107R) had no effect on transcription arrest in vitro or in vivo. Nun K106A/K107A was partially active, whereas Nun K106D/K107D was defective in vitro and failed to exclude lambda. All mutants bound RNA polymerase and BOXB. In contrast to Nun K106R/K107R and K106A/K107A, Nun K106D/K107D did not cross-link DNA template. These results suggest that transcription arrest is facilitated by electrostatic interactions between positively charged Nun residues Lys(106) and Lys(107) and negatively charged DNA phosphate groups. These may assist intercalation of Trp(108) into template. PMID- 14742437 TI - UV-induced ataxia-telangiectasia-mutated and Rad3-related (ATR) activation requires replication stress. AB - Ataxia-telangiectasia-mutated and Rad3-related (ATR) plays an essential role in the maintenance of genome integrity and cell viability. The kinase is activated in response to DNA damage and initiates a checkpoint signaling cascade by phosphorylating a number of downstream substrates including Chk1. Unlike ataxia telangiectasia-mutated (ATM), which appears to be mainly activated by DNA double strand breaks, ATR can be activated by a variety of DNA damaging agents. However, it is still unclear what triggers ATR activation in response to such diverse DNA lesions. One model proposes that ATR can directly recognize DNA lesions, while other recent data suggest that ATR is activated by a common single-stranded DNA (ssDNA) intermediate generated during DNA repair. In this study, we show that UV lesions do not directly activate ATR in vivo. In addition, ssDNA lesions created during the repair of UV damage are also not sufficient to activate the ATR dependent pathway. ATR activation is only observed in replicating cells indicating that replication stress is required to trigger the ATR-mediated checkpoint cascade in response to UV irradiation. Interestingly, H2AX appears to be required for the accumulation of ATR at stalled replication forks. Together our data suggest that ssDNA at arrested replication forks recruits ATR and initiates ATR-mediated phosphorylation of H2AX and Chk1. Phosphorylated H2AX might further facilitate ATR activation by stabilizing ATR at the sites of arrested replication forks. PMID- 14742438 TI - AMP-activated protein kinase plays a role in the control of food intake. AB - AMP-activated protein kinase (AMPK) is the downstream component of a protein kinase cascade that acts as an intracellular energy sensor maintaining the energy balance within the cell. The finding that leptin and adiponectin activate AMPK to alter metabolic pathways in muscle and liver provides direct evidence for this role in peripheral tissues. The hypothalamus is a key regulator of food intake and energy balance, coordinating body adiposity and nutritional state in response to peripheral hormones, such as leptin, peptide YY-(3-36), and ghrelin. To date the hormonal regulation of AMPK in the hypothalamus, or its potential role in the control of food intake, have not been reported. Here we demonstrate that counter regulatory hormones involved in appetite control regulate AMPK activity and that pharmacological activation of AMPK in the hypothalamus increases food intake. In vivo administration of leptin, which leads to a reduction in food intake, decreases hypothalamic AMPK activity. By contrast, injection of ghrelin in vivo, which increases food intake, stimulates AMPK activity in the hypothalamus. Consistent with the effect of ghrelin, injection of 5-amino-4-imidazole carboxamide riboside, a pharmacological activator of AMPK, into either the third cerebral ventricle or directly into the paraventricular nucleus of the hypothalamus significantly increased food intake. These results suggest that AMPK is regulated in the hypothalamus by hormones which regulate food intake. Furthermore, direct pharmacological activation of AMPK in the hypothalamus is sufficient to increase food intake. These findings demonstrate that AMPK plays a role in the regulation of feeding and identify AMPK as a novel target for anti obesity drugs. PMID- 14742440 TI - Phospholipase D is a negative regulator of proline biosynthesis in Arabidopsis thaliana. AB - Accumulation of proline has been observed in a large number of plant species in response to drought and salt stresses, suggesting a key role of this amino acid in plant stress adaptation. Upstream components of the proline biosynthesis signal transduction pathways are still poorly defined. We provide experimental evidence that phospholipase D (PLD) is involved in the regulation of proline metabolism in Arabidopsis thaliana. The application of primary butyl alcohols, which divert part of PLD-derived phosphatidic acid by transphosphatidylation, stimulated proline biosynthesis even without hyperosmotic constraints. Moreover, application of primary butyl alcohols enhanced the proline responsiveness of seedlings to mild hyperosmotic stress. These data indicate that some PLDs are negative regulators of proline biosynthesis and that plants present a higher proline responsiveness to hyperosmotic stress when this regulator is abolished. We clearly demonstrate that PLD signaling for proline biosynthesis is similar to RD29A gene expression and different from the abscisic acid-dependent RAB18 gene expression. Our data reveal that PLDs play positive and negative roles in hyperosmotic stress signal transduction in plants, contributing to a precise regulation of ion homeostasis and plant salt tolerance. PMID- 14742439 TI - Identification of a novel cyclic AMP-response element (CRE-II) and the role of CREB-1 in the cAMP-induced expression of the survival motor neuron (SMN) gene. AB - Spinal muscular atrophy, an autosomal recessive disorder, is caused by loss of the SMN1 (survival motor neuron) gene while retaining the SMN2 gene. SMN1 produces a majority of full-length SMN transcript, whereas SMN2 generates mostly an isoform lacking exon 7. Here, we demonstrate a novel cAMP-response element, CRE-II, in the SMN promoter that interacts with the cAMP-response element-binding (CREB) family of proteins. In vitro DNase I protection analysis and in vivo genomic footprinting of the SMN promoter using the brain and liver nuclei from SMN2 transgenic mice revealed footprinting at the CRE-II site. Site-directed mutation of the CRE-II element caused a marked reduction in the SMN promoter activity revealed by transient transfection assay. Activation of the cAMP pathway by dibutyryl cAMP (0.5 mm) alone or in combination with forskolin (20 microm) caused a 2-5-fold increase in the SMN promoter activity but had no effect on the CRE-II mutated promoter. Electrophoretic mobility shift assay and a UV-induced DNA-protein cross-linking experiment confirmed that CREB1 binds specifically to the CRE-II site. Transient overexpression of CREB1 protein resulted in a 4-fold increase of the SMN promoter activity. Intraperitoneal injection of epinephrine in mice expressing two copies of the human SMN2 gene resulted in a 2-fold increase in full-length SMN transcript in the liver. Combined treatment with dibutyryl cAMP and forskolin significantly increased the level of both the full length and exon 7-deleted SMN (exonDelta7SMN) transcript in primary hepatocytes from mice expressing two copies of human SMN2 gene. Similar treatments of type I spinal muscular atrophy mouse and human fibroblasts as well as HeLa cells resulted in an augmented level of SMN transcript. These findings suggest that the CRE-II site in SMN promoter positively regulates the expression of the SMN gene, and treatment with cAMP-elevating agents increases expression of both the full length and exonDelta7SMN transcript. PMID- 14742441 TI - Involvement of calcineurin in transforming growth factor-beta-mediated regulation of extracellular matrix accumulation. AB - Calcineurin is a calcium-dependent, serine/threonine phosphatase that functions as a signaling intermediate. In this study, we investigated the role of calcineurin in transforming growth factor-beta (TGF-beta)-mediated cellular effects and examined the signaling pathway involved in activation of calcineurin. Calcineurin is activated by TGF-beta in a time- and dose-dependent manner. Consistent with increased phosphatase activity, the calcineurin substrate, NFATc1, is dephosphorylated and transported to the nucleus. Inhibition of calcineurin prior to the addition of TGF-beta revealed that calcineurin is required for TGF-beta-mediated accumulation of extracellular matrix (ECM) proteins but not cell hypertrophy. Conversely, overexpression of constitutively active calcineurin was sufficient to induce ECM protein expression. The mechanism of calcineurin activation by TGF-beta was found to be induction of a low, sustained increase of intracellular calcium. Chelation of extracellular calcium blocked both TGF-beta-mediated calcium influx and calcineurin activity. Finally, calcium entry was found to be dependent upon generation of reactive oxygen species (ROS) including superoxide anion and hydrogen peroxide. Accordingly, inhibition of ROS generation also blocked TGF-beta-mediated calcineurin phosphatase activity and decreased ECM accumulation. In conclusion, this study describes a new pathway for TGF-beta-mediated regulation of ECM via generation of ROS, calcium influx, and activation of calcineurin. PMID- 14742442 TI - Analysis of SOCS-3 promoter responses to interferon gamma. AB - SOCS-3 (suppressor of cytokine signaling 3) is an intracellular protein that is selectively and rapidly induced by appropriate agonists and that modulates responses of immune cells to cytokines by interfering with the Janus kinase/signal transducer and activator of transcription (Jak/STAT) pathway. On the basis of the observations that interferon gamma (IFNgamma) up-regulates SOCS 3 gene and protein expression in primary mouse macrophages, J774 macrophage cell line and embryonal fibroblasts, we investigated which sequences of the 5' SOCS-3 gene are responsive to IFNgamma. By promoter deletion analysis we identified a functional IFNgamma-responsive element, located at nucleotides -72/-64 upstream from the transcription initiation, whose presence and integrity is necessary to ensure responsiveness to IFNgamma. This element contains a STAT consensus binding sequence (SOCS-3/STAT-binding element (SBE)) whose specific mutation totally abolished the responsiveness to IFNgamma. In contrast, discrete deletion of other 5' regions of the SOCS-3 promoter did not substantially modify the inducibility by IFNgamma. Electromobility shift assay analyses revealed that IFNgamma promotes specific DNA binding activities to an oligonucleotide probe containing the SOCS 3/SBE sequence. Even though IFNgamma triggered tyrosine phosphorylation of both STAT1 and STAT3 in macrophages and J774 cells, only STAT1 was appropriately activated and thus found to specifically bind to the SOCS-3/SBE oligonucleotide probe. Accordingly, IFNgamma-induced SOCS-3 protein expression was not impaired in STAT3-deficient embryonal fibroblasts. Taken together, these results demonstrate that the induction of SOCS-3 by IFNgamma depends upon the presence of a STAT-binding element in the SOCS-3 promoter that is specifically activated by STAT1. PMID- 14742443 TI - Suppression of WEE1 and stimulation of CDC25A correlates with endothelin dependent proliferation of rat aortic smooth muscle cells. AB - Proliferation of vascular smooth muscle cells plays a key role in the pathogenesis of several disorders of the vascular wall. Endothelin (ET), a vasoactive peptide that signals through a G protein-coupled receptor, has been linked to mitogenesis in vascular smooth muscle cells, but the mechanistic details underlying this activity remain incompletely understood. In the present study, we demonstrate that ET-dependent mitogenesis in rat neonatal and adult aortic smooth muscle (RASM) cells is accompanied by an increase (up to 10-fold) in CDK2 activity, but not CDK2 protein levels. This effect is blocked almost entirely by PD98059 and UO126, implying involvement of the MEK/ERK signal transduction cascade in the activation. Extracts of ET-treated cells phosphorylate the N terminus of WEE1, an inhibitory kinase, which negatively regulates CDK2 activity through phosphorylation at Tyr(15), leading to a decrease in WEE1 activity and a reduction in levels of phospho-Tyr(15) in the CDK2 protein. ET also increases expression and activity of CDC25A, the regulatory phosphatase responsible for dephosphorylating Tyr(15). All of these effects are reversible following treatment with the MEK inhibitor PD98059. ET also increases levels of CDC2 activity in these cells in association with a decrease in levels of phospho-Tyr(15) on the CDC2 molecule. Phosphorylation of WEE1 is linked to ERK while phosphorylation of MYT1 (CDC2-selective inhibitory kinase) is tied to the ribosomal S6 kinase (RSK). In summary, ET controls progression through the cell cycle, in part, by increasing CDK2 and CDC2 activity through the MEK/ERK/RSK signal transduction pathway(s). This results from the phosphorylation and subsequent inactivation of two inhibitory kinases (WEE1 and MYT1) that tonically suppress CDK2 and CDC2 activity and activation of a phosphatase (CDC25A) that increases CDK2 activity. PMID- 14742444 TI - Mlx is the functional heteromeric partner of the carbohydrate response element binding protein in glucose regulation of lipogenic enzyme genes. AB - The expression of genes encoding enzymes involved in de novo triglyceride synthesis (lipogenesis) is transcriptionally induced in the liver in response to increased glucose metabolism. The carbohydrate response element-binding protein (ChREBP) is a newly identified basic helix-loop-helix/leucine zipper transcription factor proposed to regulate the expression of the glucose responsive gene pyruvate kinase. This gene contains a carbohydrate response element (ChoRE) consisting of two E box motifs separated by 5 bp that is necessary and sufficient for glucose regulation. We demonstrate that overexpression of ChREBP in primary rat hepatocytes activates other ChoRE containing promoters in a manner consistent with their ability to respond to glucose. In vitro binding of ChREBP to ChoRE sequences was not detected. Because E box-binding proteins function as obligate dimers, we performed a yeast two hybrid screen of a mouse liver cDNA library to identify potential heteromeric partners. Mlx (Max-like protein X) was selected as the only basic helix-loop helix/leucine zipper interaction partner in this screen. When a plasmid expressing either Mlx or ChREBP was cotransfected with a ChoRE-containing reporter plasmid into human embryonic kidney 293 cells, no increase in promoter activity was observed. However, the expression of both proteins dramatically enhanced promoter activity. This activation was observed with reporters containing ChoREs from several different lipogenic enzyme genes. In contrast, reporters containing non-glucose-responsive E box elements were not activated by ChREBP-Mlx expression. In vitro binding of ChREBP to ChoRE-containing oligonucleotides was observed only in the presence of Mlx. ChREBP-Mlx binding discriminated between E box sites that are glucose-responsive and those that are not. We conclude that Mlx is a functional heteromeric partner of ChREBP in regulating the expression of glucose-responsive genes. PMID- 14742445 TI - Inhibition of lens fiber cell morphogenesis by expression of a mutant SV40 large T antigen that binds CREB-binding protein/p300 but not pRb. AB - Simian virus (SV) 40 large T antigen can both induce tumors and inhibit cellular differentiation. It is not clear whether these cellular changes are synonymous, sequential, or distinct responses to the protein. T antigen is known to bind to p53, to the retinoblastoma (Rb) family of tumor suppressor proteins, and to other cellular proteins such as p300 family members. To test whether SV40 large T antigen inhibits cellular differentiation in vivo in the absence of cell cycle induction, we generated transgenic mice that express in the lens a mutant version of the early region of SV40. This mutant, which we term E107KDelta, has a deletion that eliminates synthesis of small t antigen and a point mutation (E107K) that results in loss of the ability to bind to Rb family members. At embryonic day 15.5 (E15.5), the transgenic lenses show dramatic defects in lens fiber cell differentiation. The fiber cells become post-mitotic, but do not elongate properly. The cells show a dramatic reduction in expression of their beta- and gamma-crystallins. Because CBP and p300 are co-activators for crystallin gene expression, we assayed for interactions between E107KDelta and CBP/p300. Our studies demonstrate that cellular differentiation can be inhibited by SV40 large T antigen in the absence of pRb inactivation, and that interaction of large T antigen with CBP/p300 may be enhanced by a mutation that eliminates the binding to pRb. PMID- 14742447 TI - Glycogen synthase sensitivity to glucose-6-P is important for controlling glycogen accumulation in Saccharomyces cerevisiae. AB - Glycogen is a storage form of glucose utilized as an energy reserve by many organisms. Glycogen synthase, which is essential for synthesizing this glucose polymer, is regulated by both covalent phosphorylation and the concentration of glucose-6-P. With the yeast glycogen synthase Gsy2p, we recently identified two mutants, R579A/R580A/R582A [corrected] and R586A/R588A/R591A, in which multiple arginine residues were mutated to alanine that were completely insensitive to activation by glucose-6-P in vitro (Pederson, B. A., Cheng, C., Wilson, W. A., and Roach, P. J. (2000) J. Biol. Chem. 275, 27753-27761). We report here the expression of these mutants in Saccharomyces cerevisiae and, as expected from our findings in vitro, they were not activated by glucose-6-P. The R579A/R580A/R582A [corrected] mutant, which is also resistant to inhibition by phosphorylation, caused hyperaccumulation of glycogen. In contrast, the mutant R586A/R588A/R591A, which retains the ability to be inactivated by phosphorylation, resulted in lower glycogen accumulation when compared with wild-type cells. When intracellular glucose-6-P levels were increased by mutating the PFK2 gene, glycogen storage due to the wild-type enzyme was increased, whereas that associated with R579A/R580A/R582A [corrected] was not greatly changed. This is the first direct demonstration that activation of glycogen synthase by glucose-6-P in vivo is necessary for normal glycogen accumulation. PMID- 14742446 TI - Calcium dependence of polycystin-2 channel activity is modulated by phosphorylation at Ser812. AB - Polycystin-2 (PC-2) is a non-selective cation channel that, when mutated, results in autosomal dominant polycystic kidney disease. In an effort to understand the regulation of this channel, we investigated the role of protein phosphorylation in PC-2 function. We demonstrated the direct incorporation of phosphate into PC-2 in cells and tissues and found that this constitutive phosphorylation occurs at Ser(812), a putative casein kinase II (CK2) substrate domain. Ser(812) can be phosphorylated by CK2 in vitro and substitution S812A results in failure to incorporate phosphate in cultured epithelial cells. Non-phosphorylated forms of PC-2 traffic normally in the endoplasmic reticulum and cilial compartments and retain homo- and hetero-multimerization interactions with PC-2 and polycystin-1, respectively. Single-channel studies of PC-2, S812A, and a substitution mutant, T721A, not related to phosphorylation show that PC-2 and S812A function as divalent cation channels with similar current amplitudes across a range of holding potentials; the T721A channel is not functional. Channel open probabilities for PC-2 and S812A show a bell-shaped dependence on cytoplasmic Ca(2+) but there is a shift in this Ca(2+) dependence such that S812A is 10-fold less sensitive to Ca(2+) activation/inactivation than the wild type PC-2 channel. In vivo analysis of PC-2-dependent enhanced intracellular Ca(2+) transients found that S812A resulted in enhanced transient duration and relative amplitude intermediate between control cells and those overexpressing wild type PC-2. Phosphorylation at Ser(812) modulates PC-2 channel activity and factors regulating this phosphorylation are likely to play a role in the pathogenesis of polycystic kidney disease. PMID- 14742448 TI - Alpha-synuclein up-regulation and aggregation during MPP+-induced apoptosis in neuroblastoma cells: intermediacy of transferrin receptor iron and hydrogen peroxide. AB - 1-Methyl-4-phenylpyridinium (MPP(+)) is a neurotoxin that causes Parkinson's disease in experimental animals and humans. Despite the fact that intracellular iron was shown to be crucial for MPP(+)-induced apoptotic cell death, the molecular mechanisms for the iron requirement remain unclear. We investigated the role of transferrin receptor (TfR) and iron in modulating the expression of alpha synuclein (alpha-syn) in MPP(+)-induced oxidative stress and apoptosis. Results show that MPP(+) inhibits mitochondrial complex-1 and aconitase activities leading to enhanced H(2)O(2) generation, TfR expression and alpha-syn expression/aggregation. Pretreatment with cell-permeable iron chelators, TfR antibody (that inhibits TfR-mediated iron uptake), or transfection with glutathione peroxidase (GPx1) enzyme inhibits intracellular oxidant generation, alpha-syn expression/aggregation, and apoptotic signaling as measured by caspase 3 activation. Cells overexpressing alpha-syn exacerbated MPP(+) toxicity, whereas antisense alpha-syn treatment totally abrogated MPP(+)-induced apoptosis in neuroblastoma cells without affecting oxidant generation. The increased cytotoxic effects of alpha-syn in MPP(+)-treated cells were attributed to inhibition of mitogen-activated protein kinase and proteasomal function. We conclude that MPP(+)-induced iron signaling is responsible for intracellular oxidant generation, alpha-syn expression, proteasomal dysfunction, and apoptosis. Relevance to Parkinson's disease is discussed. PMID- 14742449 TI - Hypoxia-induced activation of the retinoic acid receptor-related orphan receptor alpha4 gene by an interaction between hypoxia-inducible factor-1 and Sp1. AB - Hypoxia plays a key role in the pathophysiology of many disease states, and expression of the retinoic acid receptor-related orphan receptor alpha (RORalpha) gene increases under hypoxia. We investigated the mechanism for this transient hypoxia-induced increase in RORalpha expression. Reverse transcription-coupled PCR analysis revealed that the steady-state level of mRNA for the RORalpha4 isoform, but not the RORalpha1 isoform, increased in HepG2 cells after 3 h of hypoxia. Transient transfection studies showed that the hypoxia-induced increase in RORalpha4 mRNA occurs at the transcriptional level and is dependent on a hypoxia-responsive element (HRE) located downstream of the promoter. A dominant negative mutant of hypoxia-inducible factor-1alpha (HIF-1alpha) abrogates the transcription activated by hypoxia as well as the transcription activated by exogenously expressed HIF-1alpha, demonstrating the direct involvement of HIF 1alpha in the transcriptional activation. However, HIF-1 alone was not sufficient to activate transcription in hypoxic conditions but, rather, required Sp1/Sp3, which binds to a cluster of GC-rich sequences adjacent to the HRE. Deletion of one or more of these GC boxes reduced or eliminated the HIF-1-dependent transcription. Together, these results suggest that the hypoxia-responsive region of the RORalpha4 promoter is composed of the HRE and GC-rich sequences and that the transcriptional activation under hypoxia is conferred through the cooperation of HIF-1 with Sp1/Sp3. PMID- 14742450 TI - A genetic polymorphism of the alpha2-adrenergic receptor increases autonomic responses to stress. AB - We hypothesized that individual differences in autonomic responses to psychological, physiological, or environmental stresses are inherited, and exaggerated autonomic responsiveness may represent an intermediate phenotype that can contribute to the development of essential hypertension in humans over time. alpha(2)-Adrenergic receptors (alpha(2)-ARs), encoded by a gene on chromosome 10, are found in the central nervous system and also mediate release of norepinephrine from the presynaptic nerve terminals of the peripheral sympathetic nervous system and the exocytosis of epinephrine from the adrenal medulla. We postulated that, because this receptor mediates central and peripheral autonomic responsiveness to stress, genetic mutations in the gene encoding this receptor may explain contrasting activity of the autonomic nervous system among individuals. The restriction enzyme Dra I identifies a polymorphic site in the 3' transcribed, but not translated, portion of the gene encoding the chromosome 10 alpha(2)-AR. Southern blotting of genomic DNA with a cDNA probe after restriction enzyme digestion results in fragments that are either 6.7 kb or 6.3 kb in size. Transfection studies of these two genotypes resulted in contrasting expression of a reporter gene, and it is suggested from these findings that this is a functional polymorphism. In a study of 194 healthy subjects, we measured autonomic responses to provocative motion, a fall in blood pressure induced by decreasing venous return and cardiac output, or exercise. Specifically, we measured reactions to 1) Coriolis stress, a strong stimulus that induces motion sickness in man; 2) heart rate responses to the fall in blood pressure induced by the application of graded lower body negative pressure; and 3) exercise-induced sweat secretion. In all of these paradigms of stress, subjective and objective evidence of increased autonomic responsiveness was found in those individuals harboring the 6.3-kb allele. Specifically, volunteers with the 6.3-kb allele had greater signs and symptoms of motion sickness mediated by the autonomic nervous system after off-axis rotation at increasing velocity (number of head movements a subject could complete during rotation before emesis +/- SE: 295 +/- 18 vs. 365 +/- 11; P = 0.001). They also had greater increases in heart rate in responses to the lower body negative pressure-induced fall in blood pressure (increase in heart rate +/- SE: 3.0 +/- 0.4 vs. 1.8 +/- 0.3; P = 0.012), and the 6.3-kb group had higher sweat sodium concentrations during exercise (mean sweat sodium concentration in meq/l over 30 min of exercise +/- SE: 43.2 +/- 7.1 vs. 27.6 +/- 3.4; P < 0.05). This single-nucleotide polymorphism may contribute to contrasting individual differences in autonomic responsiveness among healthy individuals. PMID- 14742451 TI - Longer static flexion duration elicits a neuromuscular disorder in the lumbar spine. AB - The objective of this study was to assess the impact of two sequential long, static, anterior lumbar flexions on the development of a neuromuscular disorder and to compare it with previously obtained data from a series of short static flexion periods of the same cumulative time (Sbriccoli P, Solomonow M, Zhou BH, Baratta RV, Lu Y, Zhu MP, and Burger EL, Muscle Nerve 29: 300-308, 2004). Static flexions with loads of 20, 40, and 60 N were applied to the lumbar spine over two 30-min periods with a 10-min rest in between. The reflex EMG activity from the multifidus muscles and supraspinous ligament displacement (creep) was recorded during the flexion periods. Creep and EMG were also monitored over 7 h of rest following the work-rest-work cycle. It was found that the creep that developed in the first 30-min flexion period did not recover completely during the following 10 min of rest, giving rise to a large cumulative creep at the end of the work rest-work session. Spasms were frequently seen within the EMG during the static flexion. Initial and delayed hyperexcitabilities were observed in all of the preparations at any of the three loads explored during the 7-h rest period. ANOVA revealed a significant effect of time (P < 0.0001) on the postloading data. Larger loads elicited larger magnitudes of the initial and delayed hyperexcitabilities, yet were not statistically different. It was concluded that the 3:1 work-to-rest duration ratio resulted in a neuromuscular disorder, regardless of the load magnitude. The conclusions are reinforced in view of the results from a previous study using 60 min of flexion overall but at 1:1 work-to rest ratio in which only the highest load elicited a delayed hyperexcitability (Sbriccoli et al., Muscle Nerve 29: 300-308, 2004). An optimal dose-to-duration ratio needs to be established to limit, attenuate, or prevent the adverse effects of static load on the lumbar spine while considering the loading duration as a major risk factor. PMID- 14742452 TI - Validation of publication of new names and new combinations previously effectively published outside the IJSEM. AB - The purpose of this announcement is to effect the valid publication of the following new names and new combinations under the procedure described in the Bacteriological Code (1990 Revision). Authors and other individuals wishing to have new names and/or combinations included in future lists should send three copies of the pertinent reprint or photocopies thereof to the IJSEM Editorial Office for confirmation that all of the other requirements for valid publication have been met. It is also a requirement of IJSEM and the ICSP that authors of new species, new subspecies and new combinations provide evidence that types are deposited in two recognized culture collections in two different countries (i.e. documents certifying deposition and availability of type strains). It should be noted that the date of valid publication of these new names and combinations is the date of publication of this list, not the date of the original publication of the names and combinations. The authors of the new names and combinations are as given below, and these authors' names will be included in the author index of the present issue and in the volume author index. Inclusion of a name on these lists validates the publication of the name and thereby makes it available in bacteriological nomenclature. The inclusion of a name on this list is not to be construed as taxonomic acceptance of the taxon to which the name is applied. Indeed, some of these names may, in time, be shown to be synonyms, or the organisms may be transferred to another genus, thus necessitating the creation of a new combination. PMID- 14742453 TI - Exploring prokaryotic taxonomy. AB - Techniques drawn from exploratory data analysis, using tools found in the S-Plus statistical software package, have been used to inspect and maintain the Bergey's Taxonomic Outline and to move towards an automated and community-based means of working on the outline. These techniques can be used to classify sequences from unnamed and uncultured organisms, to visualize errors in the taxonomy or in the curation of the sequences, to suggest emendations to the taxonomy or to the classification of extant species and to complement the visualization of phylogenies based on treeing methods. A dataset of more than 9200 aligned small subunit rRNA sequences was analysed in the context of the current taxonomic outline. The use of the algorithm in exploring and modifying the taxonomy is illustrated with an example drawn from the family Comamonadaceae. PMID- 14742454 TI - Polyphasic characterization of xanthomonads isolated from onion, garlic and Welsh onion (Allium spp.) and their relatedness to different Xanthomonas species. AB - Bacterial blight is an emerging disease that affects primarily onion, but also garlic and Welsh onion. The present study was undertaken to characterize the causative xanthomonad(s) by a polyphasic approach using a worldwide collection of 33 bacterial strains. Analysis of 16S rRNA gene sequence similarities indicated that the causal agent belongs to the campestris core in the genus Xanthomonas, which is in agreement with results of phenotypic characterization (analyses of carbon source utilization and fatty acid methyl esters). However, DNA-DNA hybridization, thermal stability of DNA reassociation and fluorescent amplified fragment length polymorphism analysis allowed the causal agent to be identified as a pathovar of Xanthomonas axonopodis. PMID- 14742455 TI - Hydrogenimonas thermophila gen. nov., sp. nov., a novel thermophilic, hydrogen oxidizing chemolithoautotroph within the epsilon-Proteobacteria, isolated from a black smoker in a Central Indian Ridge hydrothermal field. AB - A novel thermophilic bacterium, strain EP1-55-1%T, was isolated from an in-situ colonization system deployed in a superheated, deep-sea, hydrothermal vent emission at the Kairei Field on the Central Indian Ridge in the Indian Ocean. The cells were highly motile rods, each possessing a single polar flagellum. Growth was observed between 35 and 65 degrees C (optimum temperature, 55 degrees C; 70 min doubling time) and between pH 4.9 and 7.2 (optimum, pH 5.9). The isolate was a microaerobic-to-anaerobic chemolithoautotroph capable of using molecular hydrogen as the sole energy source and carbon dioxide as the sole carbon source. Molecular oxygen, nitrate or elemental sulfur (S0) could serve as electron acceptors to support growth. The G+C content of the genomic DNA was 34.6 mol%. Phylogenetic analysis based on 16S rDNA sequences indicated that strain EP1-55 1%T represents the first strain for which taxonomic properties have been characterized within the previously uncultivated phylogroup classified as belonging to the uncultivated epsilon-Proteobacteria group A; the name Hydrogenimonas thermophila gen. nov., sp. nov. is proposed, with strain EP1-55 1%T (=JCM 11971T=ATCC BAA-737T) as the type strain. PMID- 14742456 TI - Sulfurihydrogenibium azorense, sp. nov., a thermophilic hydrogen-oxidizing microaerophile from terrestrial hot springs in the Azores. AB - Five hydrogen-oxidizing, thermophilic, strictly chemolithoautotrophic, microaerophilic strains, with similar (99-100%) 16S rRNA gene sequences were isolated from terrestrial hot springs at Furnas, Sao Miguel Island, Azores, Portugal. The strain, designated Az-Fu1T, was characterized. The motile, 0.9-2.0 microm rods were Gram-negative and non-sporulating. The temperature growth range was from 50 to 73 degrees C (optimum at 68 degrees C). The strains grew fastest in 0.1% (w/v) NaCl and at pH 6, although growth was observed from pH 5.5 to 7.0. Az-Fu1T can use elemental sulfur, sulfite, thiosulfate, ferrous iron or hydrogen as electron donors, and oxygen (0.2-9.0%, v/v) as electron acceptor. Az-Fu1T is also able to grow anaerobically, with elemental sulfur, arsenate and ferric iron as electron acceptors. The Az-Fu1T G+C content was 33.6 mol%. Maximum-likelihood analysis of the 16S rRNA phylogeny placed the isolate in a distinct lineage within the Aquificales, closely related to Sulfurihydrogenibium subterraneum (2.0% distant). The 16S rRNA gene of Az-Fu1T is 7.7% different from that of Persephonella marina and 6.8% different from Hydrogenothermus marinus. Based on the phenotypic and phylogenetic characteristics presented here, it is proposed that Az-Fu1T belongs to the recently described genus Sulfurihydrogenibium. It is further proposed that Az-Fu1T represents a new species, Sulfurihydrogenibium azorense. PMID- 14742457 TI - Caminibacter profundus sp. nov., a novel thermophile of Nautiliales ord. nov. within the class 'Epsilonproteobacteria', isolated from a deep-sea hydrothermal vent. AB - A novel moderately thermophilic, microaerobic to anaerobic, chemolithoautotrophic bacterium, designated strain CRT, was isolated from a deep-sea hydrothermal vent site at 36 degrees N on the Mid-Atlantic Ridge. Cells were Gram-negative, non motile rods. The organism grew at 45-65 degrees C and pH 6.5-7.4, with optimum growth at 55 degrees C and pH 6.9-7.1. The NaCl range for growth was 5-50 g l(-1) (optimum 30 g l(-1)). Strain CRT was an obligate chemolithoautotroph, growing with H2 as energy source, sulfur, nitrate or oxygen as electron acceptors and CO2 as carbon source. Hydrogen sulfide and ammonium were the respective products of sulfur and nitrate reduction. The G+C content of the genomic DNA was 32.1 mol%. Based on 16S rRNA gene sequence analysis, this organism was most closely related to Caminibacter hydrogeniphilus (94.9% similarity). On the basis of phenotypic and phylogenetic data, it is proposed that the isolate represents a novel species, Caminibacter profundus sp. nov. The type strain is CRT (=DSM 15016T=JCM 11957T). The phylogenetic data also correlate well with the significant phenotypic differences between the lineage encompassing the genera Nautilia and Caminibacter and other members of the class 'Epsilonproteobacteria'. The lineage encompassing the genera Nautilia and Caminibacter is therefore proposed as a new order, Nautiliales ord. nov., represented by a single family, Nautiliaceae fam. nov. PMID- 14742458 TI - Bacillus novalis sp. nov., Bacillus vireti sp. nov., Bacillus soli sp. nov., Bacillus bataviensis sp. nov. and Bacillus drentensis sp. nov., from the Drentse A grasslands. AB - A group of 42 isolates were isolated from the soil of several disused hay fields, in the Drentse A agricultural research area (The Netherlands), that were taken out of production at different times. The group represents hitherto-uncultured Bacillus lineages that have previously been found, by a non-cultural method, to be predominant in soil. The strains were subjected to a polyphasic taxonomic study, including (GTG)5-PCR, 16S rDNA sequence analysis, DNA-DNA hybridizations, DNA base-ratio determination, fatty acid analysis and morphological and biochemical characterization. By comparing the groupings obtained by (GTG)5-PCR and 16S rDNA sequence analysis, six clusters of similar strains could be recognized. A DNA-DNA relatedness study showed that these clusters represented five novel genospecies. Further analysis supported the proposal of five novel species in the genus Bacillus, namely Bacillus novalis sp. nov. (type strain IDA3307T=R-15439T=LMG 21837T=DSM 15603T), Bacillus vireti sp. nov. (type strain IDA3632T=R-15447T=LMG 21834T=DSM 15602T), Bacillus soli sp. nov. (type strain IDA0086T=R-16300T=LMG 21838T=DSM 15604T), Bacillus bataviensis sp. nov. (type strain IDA1115T=R-16315T=LMG 21833T=DSM 15601T) and Bacillus drentensis sp. nov. (type strain IDA1967T=R-16337T=LMG 21831T=DSM 15600T). PMID- 14742459 TI - Paenibacillus favisporus sp. nov., a xylanolytic bacterium isolated from cow faeces. AB - During a search for xylan-degrading micro-organisms, a sporulated bacterium was recovered from recent and old cow dung and rectal samples. The isolates were identified as members of a novel species of the genus Paenibacillus, based on 16S rRNA gene sequences. According to the results of phylogenetic analysis, the most closely related species was Paenibacillus azoreducens. Phenotypic and chemotaxonomic analyses and DNA-DNA hybridization experiments also showed that the isolates belonged to a novel species of the genus Paenibacillus. The novel species is a facultatively anaerobic, motile, Gram-variable, sporulated rod. The spores of this rod-shaped micro-organism occur in slightly swollen sporangia and are honeycomb-shaped. The main fatty acid is anteiso-branched C(15:0). Growth was observed with many carbohydrates, including xylan, as the only carbon source and gas production was not observed from glucose. The novel species produces a wide variety of hydrolytic enzymes, such as xylanases, cellulases, amylases, gelatinase, urease and beta-galactosidase. On the contrary, it does not produce caseinase, phenylalanine deaminase or lysine decarboxylase. According to the data obtained in this work, the strains belong to a novel species, for which the name Paenibacillus favisporus sp. nov. is proposed (type strain, GMP01T=LMG 20987T=CECT 5760T). PMID- 14742460 TI - Belliella baltica gen. nov., sp. nov., a novel marine bacterium of the Cytophaga Flavobacterium-Bacteroides group isolated from surface water of the central Baltic Sea. AB - Two bacterial isolates from the Baltic Sea, BA1 and BA134T, were characterized for their physiological and biochemical features, fatty acid profiles and phylogenetic position based on 16S rRNA gene sequences. The strains were isolated from surface water of the central Baltic Sea during the decay of a plankton bloom. Phylogenetic analysis of their 16S rRNA gene sequences revealed a clear affiliation to the family 'Flexibacteriaceae' and showed highest sequence similarity (91%) to Cyclobacterium marinum. The G+C content of the DNA was 35.4 mol%. The strains were pink-coloured due to carotinoids, Gram-negative, rod shaped and catalase- and oxidase-positive. Growth was observed at 0-6% salinity, with good growth at 0-3%. Temperature for growth was 4-37 degrees C, with an optimum around 25 degrees C. The fatty acid profiles were dominated by branched chain fatty acids (70%), with a high abundance of iso-C(15:0) (29-33%), iso C(17:1)omega9c (7-10%) and C(17:1)omega6c (5-10%). According to their morphology, physiology, fatty acid composition, 16S rRNA gene sequences and DNA-DNA similarity, on one hand, the described bacteria are considered to be members of the same novel species; on the other hand, they are suggested as a novel genus of the family 'Flexibacteriaceae'. To honour the late aquatic microbiologist Russell T. Bell, the name Belliella baltica gen. nov, sp. nov. is suggested for the Baltic Sea isolates, for which the type strain is BA134T (=DSM 15883T=LMG 21964T=CIP 108006T). PMID- 14742461 TI - Nocardioides aquiterrae sp. nov., isolated from groundwater in Korea. AB - A bacterial strain, GW-9T, which was isolated from groundwater in Korea, was subjected to a polyphasic taxonomic study using phenotypic characterization and phylogenetic and genetic methods. Phylogenetic analysis based on 16S rDNA sequences showed that strain GW-9T forms an evolutionary lineage within the radiation enclosing Nocardioides species and, in particular, a coherent cluster with Nocardioides pyridinolyticus. The cell-wall peptidoglycan type of strain GW 9T was based on LL-diaminopimelic acid as the diamino acid, indicating wall chemotype I. The predominant menaquinone was MK-8(H4). Strain GW-9T had a cellular fatty acid profile containing straight-chain, branched, unsaturated and 10-methyl fatty acids. The major fatty acid was iso-C(16:0). The DNA G+C content of strain GW-9T was 73 mol%. The 16S rDNA of strain GW-9T was 99.2% similar to that of the type strain of Nocardioides pyridinolyticus and 94.9-96.0% similar to sequences of the type strains of other Nocardioides species. Differences in phenotypic characteristics and genetic distinctiveness indicate that strain GW-9T is separate from previously described Nocardioides species. Therefore, on the basis of the data presented, a novel species of the genus Nocardioides, Nocardioides aquiterrae sp. nov., is proposed. The type strain is strain GW-9T (=KCCM 41647T=JCM 11813T). PMID- 14742462 TI - Desulfatibacillum aliphaticivorans gen. nov., sp. nov., an n-alkane- and n-alkene degrading, sulfate-reducing bacterium. AB - A novel marine sulfate-reducing bacterium, strain CV2803T, which is able to oxidize aliphatic hydrocarbons, was isolated from a hydrocarbon-polluted marine sediment (Gulf of Fos, France). The cells were rod-shaped and slightly curved, measuring 0.6x2.2-5.5 microm. Strain CV2803T stained Gram-negative and was non motile and non-spore-forming. Optimum growth occurred in the presence of 24 g NaCl l(-1), at pH 7.5 and at a temperature between 28 and 35 degrees C. Strain CV2803T oxidized alkanes (from C13 to C18) and alkenes (from C7 to C23). The DNA G+C content was 41.4 mol%. Comparative sequence analyses of the 16S rRNA gene and dissimilatory sulfite reductase (dsrAB) gene and those of other sulfate-reducing bacteria, together with its phenotypic properties, indicated that strain CV2803T was a member of a distinct cluster that contained unnamed species. Therefore, strain CV2803T (=DSM 15576T=ATCC BAA-743T) is proposed as the type strain of a novel species in a new genus, Desulfatibacillum aliphaticivorans gen. nov., sp. nov. PMID- 14742463 TI - Flavobacterium degerlachei sp. nov., Flavobacterium frigoris sp. nov. and Flavobacterium micromati sp. nov., novel psychrophilic bacteria isolated from microbial mats in Antarctic lakes. AB - Taxonomic studies were performed on 36 strains that were isolated from microbial mats in Antarctic lakes of the Vestfold Hills, the Larsemann Hills and the McMurdo Dry Valleys. Phylogenetic analysis based on 16S rRNA gene sequences indicated that these strains are related to members of the genus Flavobacterium; sequence similarity values with their nearest phylogenetic neighbours ranged from 96.8 to 98.5%. Results of DNA-DNA hybridization and comparison of repetitive extragenic palindromic DNA-PCR fingerprinting patterns revealed that these strains are members of three distinct species. Genotypic results, together with phenotypic characteristics, allowed the differentiation of these species from related Flavobacterium species with validly published names. The isolates are Gram-negative, chemoheterotrophic, rod-shaped cells that are psychrophilic and moderately halotolerant; their DNA G+C contents range from 33.1 to 34.5 mol%. Their whole-cell fatty acid profiles are similar and include C(15:0), anteiso C(15:0), iso-C(15:0), C(15:1)omega6c, iso-C(16:0), iso-C(16:0) 3-OH and summed feature 3 (which comprises iso-C(15:0) 2-OH, C(16:1)omega7c or both) as major fatty acid components. On the basis of these results, three novel species are proposed, namely Flavobacterium degerlachei sp. nov. (consisting of 14 strains, with LMG 21915T=DSM 15718T as the type strain), Flavobacterium micromati sp. nov. (consisting of three strains, with LMG 21919T=CIP 108161T as the type strain) and Flavobacterium frigoris sp. nov. (consisting of 19 strains, with LMG 21922T=DSM 15719T as the type strain). PMID- 14742464 TI - Polaromonas naphthalenivorans sp. nov., a naphthalene-degrading bacterium from naphthalene-contaminated sediment. AB - Strain CJ2T, capable of growth on naphthalene as a sole carbon and energy source, was isolated from coal-tar-contaminated freshwater sediment. The Gram reaction of strain CJ2T was negative. The cells were non-spore-forming, non-motile cocci (without flagella). The isolate was found to be an aerobic heterotroph capable of utilizing glucose and other simple sugars. Growth was observed between 4 and 25 degrees C (optimum, 20 degrees C) and between pH 6.0 and 9.0 (optimum, pH 7.0 7.5). The G+C content of the genomic DNA was 61.5 mol% and the major quinone was ubiquinone-8. The peptidoglycan of strain CJ2T was determined as belonging to type A1-gamma, meso-diaminopimelic acid. The major fatty acids of strain CJ2T were 16:1omega7c (67.0%), 16:0 (19.6%), 18:1omega7c (approximately 7.9%) and 10:0 3-OH (approximately 2.5%). The polar lipids were phosphatidylethanolamine, phosphatidylglycerol and diphosphatidylglycerol. Mycolic acid and glycolipids could not be detected. Comparative 16S rDNA analysis indicated that strain CJ2T is related to the family Comamonadaceae and that the nearest phylogenetic relative was Polaromonas vacuolata 34-PT (97.1% similarity). On the basis of the physiological and molecular properties, the naphthalene-degrading isolate was designated Polaromonas naphthalenivorans sp. nov. The type strain is CJ2T (=ATCC BAA-779T=DSM 15660T). PMID- 14742465 TI - Ottowia thiooxydans gen. nov., sp. nov., a novel facultatively anaerobic, N2O producing bacterium isolated from activated sludge, and transfer of Aquaspirillum gracile to Hylemonella gracilis gen. nov., comb. nov. AB - Strain K11T was isolated from activated sludge of a municipal wastewater treatment plant. Phylogenetic analysis of the 16S rRNA gene sequence revealed that it represents a distinct line of descent within the Comamonadaceae. The novel strain was a Gram-negative, catalase- and oxidase-positive, non-motile, straight to slightly curved rod. Polyhydroxyalkanoate granules were stored intracellularly as reserve material. Colonies on agar plates were small, regular and characterized by a water-insoluble yellow pigment. Unbranched fatty acids 16:1omega7c, 16:0 and 18:1omega7c dominated the cellular fatty acid pattern and ubiquinone-8 (Q-8) was the major component of the respiratory lipoquinones, both traits typical of members of the Comamonadaceae. A distinguishing characteristic was the presence of the two hydroxy fatty acids 10:0 3-OH and 12:0 2-OH, each in significant amounts. The G+C content of the DNA was 59 mol%. Strain K11T was capable of aerobic chemolithoheterotrophic growth using thiosulfate as an additional substrate, but could not grow autotrophically with thiosulfate or hydrogen. Facultative anaerobic growth was possible with nitrate and nitrite as electron acceptors, but not with ferric iron, sulfate or by fermentation. The sole end product of denitrification was N2O; nitrite accumulated only transiently in small amounts. Based upon phylogenetic and phenotypic evidence, it is proposed to establish the novel taxon Ottowia thiooxydans gen. nov., sp. nov., represented by the type strain K11T (=DSM 14619T=JCM 11629T). Aquaspirillum gracile was among the phylogenetically most closely related species to strain K11T. This species has been wrongly classified, and it is proposed to reclassify it as Hylemonella gracilis gen. nov., comb. nov. The type strain is ATCC 19624T (=DSM 9158T). PMID- 14742466 TI - Erwinia papayae sp. nov., a pathogen of papaya (Carica papaya). AB - Bacterial canker of papaya (Carica papaya) emerged during the 1980s in different islands of the Caribbean. Nineteen strains of Gram-negative, rod-shaped, non spore-forming bacteria isolated from papaya were compared to 38 reference and type strains of phytopathogenic Enterobacteriaceae and related bacteria. Phylogenetic analysis of 16S rRNA gene sequences showed that the papaya strains belonged to the genus Erwinia. The DNA G+C content of strain CFBP 5189T, 52.5 mol%, is in the range of the genus Erwinia. The 19 papaya strains were all pathogenic to papaya and were differentiated clearly from type or reference strains of phytopathogenic enterobacteria and related bacteria by phenotypic tests. The papaya strains constituted a discrete DNA hybridization group, indicating that they belonged to a unique genomic species. Thus, strains pathogenic to papaya belong to a novel species for which the name Erwinia papayae sp. nov. is proposed, with the type strain CFBP 5189T (=NCPPB 4294T). PMID- 14742467 TI - Lactobacillus paracollinoides sp. nov., isolated from brewery environments. AB - Three novel strains isolated from brewery environments are described. These strains were Gram-positive, facultatively anaerobic, heterofermentative rods that did not exhibit catalase activity. Phylogenetic analysis based on 16S rRNA gene sequence similarity showed that these strains belong to the genus Lactobacillus and are most closely related to Lactobacillus collinoides (approximately 99% similarity). The novel strains could be differentiated from L. collinoides on the basis of DNA-DNA relatedness, differences in beer-spoilage ability and the inability to utilize D-fructose. These isolates represent a novel species, for which the name Lactobacillus paracollinoides sp. nov. is proposed. The type strain is LA2T (=DSM 15502T=JCM 11969T). PMID- 14742468 TI - Ulvibacter litoralis gen. nov., sp. nov., a novel member of the family Flavobacteriaceae isolated from the green alga Ulva fenestrata. AB - Two heterotrophic, aerobic, Gram-negative, pigmented and non-motile marine bacteria that were isolated from the green alga Ulva fenestrata were studied by polyphasic taxonomic methods. 16S rDNA sequence analysis indicated that strain KMM 3912T formed a distinct lineage within the family Flavobacteriaceae. On the basis of phenotypic, chemotaxonomic, genotypic and phylogenetic analyses, the novel bacteria were classified as Ulvibacter litoralis gen. nov., sp. nov. The type strain is KMM 3912T (=KCTC 12104T=CCUG 47093T). PMID- 14742469 TI - Nocardia asiatica sp. nov., isolated from patients with nocardiosis in Japan and clinical specimens from Thailand. AB - Five strains isolated from two patients with nocardiosis in Japan and three clinical samples from Thailand were found to have morphological, biochemical and chemotaxonomic properties consistent with their classification in the genus Nocardia. DNA-DNA hybridization, coupled with sequence analysis of 16S rDNA, indicated that these strains belong to a novel species of the genus Nocardia, named Nocardia asiatica sp. nov. because the isolation sites were in Asian countries; the type strain is IFM 0245T (=NBRC 100129T=JCM 11892T=DSM 44668T). PMID- 14742470 TI - Pseudonocardia chloroethenivorans sp. nov., a chloroethene-degrading actinomycete. AB - A bacterial strain, SL-1T, capable of degrading trichloroethene was isolated from a laboratory enrichment in the Department of Civil and Environmental Engineering, University of Washington, USA. The material in the enrichments was derived from a soil sample from Seattle, WA, USA. Strain SL-1T was capable of using phenol as a source of carbon and energy. Chemotaxonomic, morphological, physiological and phylogenetic analyses showed that strain SL-1T is a member of the genus Pseudonocardia. The ability of strain SL-1T to utilize phenol and degrade trichloroethene, as well as other phenotypic properties and the results from a 16S rRNA phylogenetic analysis, led to the proposal of a novel species, Pseudonocardia chloroethenivorans sp. nov. The type strain is SL-1T (=ATCC BAA 742T=DSM 44698T). Trichloroethene and other chloroethenes are major pollutants at many environmental sites, and P. chloroethenivorans has biodegradation properties that should be of interest to environmental microbiologists and engineers. PMID- 14742472 TI - Renaming of Agrobacterium larrymoorei Bouzar and Jones 2001 as Rhizobium larrymoorei (Bouzar and Jones 2001) comb. nov. AB - In conformity with the nomenclature of the genus Rhizobium published by Young et al. (Int J Syst Evol Microbiol 51, 89-103, 2001), it is proposed that Agrobacterium larrymoorei be named as Rhizobium larrymoorei comb. nov. PMID- 14742471 TI - Thalassolituus oleivorans gen. nov., sp. nov., a novel marine bacterium that obligately utilizes hydrocarbons. AB - An aerobic, heterotrophic, Gram-negative, curved bacterial strain, designated MIL 1T, was isolated by extinction dilution from an n-tetradecane enrichment culture that was established from sea water/sediment samples collected in the harbour of Milazzo, Italy. In the primary enrichment, the isolate formed creamy-white, medium-sized colonies on the surface of the agar. The isolate did not grow in the absence of NaCl; growth was optimal at 2.7% NaCl. Only a narrow spectrum of organic compounds, including aliphatic hydrocarbons (C7-C20), their oxidized derivatives and acetate, were used as growth substrates. The isolate was not able to grow under denitrifying conditions. The DNA G+C content and genome size of strain MIL-1T were estimated to be 53.2 mol% and 2.2 Mbp, respectively. The major cellular and phospholipid fatty acids were palmitoleic, palmitic and oleic acids (33.5, 29.5 and 11.0% and 18, 32 and 31%, respectively). 3-hydroxy lauric acid was the only hydroxy fatty acid detected. Thirteen different compounds that belonged to two types of phospholipid (phosphatidylethylamine and phosphatidylglycerol) were identified. 16S rRNA gene sequence analysis revealed that this isolate represents a distinct phyletic lineage within the gamma Proteobacteria and has about 94.4% sequence similarity to Oceanobacter kriegii (the closest bacterial species with a validly published name). The deduced protein sequence of the putative alkane hydrolase, AlkB, of strain MIL-1T is related to the corresponding enzymes of Alcanivorax borkumensis and Pseudomonas oleovorans (81 and 80% similarity, respectively). On the basis of the analyses performed, Thalassolituus oleivorans gen. nov., sp. nov. is described. Strain MIL 1T (=DSM 14913T=LMG 21420T) is the type and only strain of T. oleivorans. PMID- 14742473 TI - Methylocella tundrae sp. nov., a novel methanotrophic bacterium from acidic tundra peatlands. AB - A novel species, Methylocella tundrae, is proposed for three methanotrophic strains (T4T, TCh1 and TY1) isolated from acidic Sphagnum tundra peatlands. These strains are aerobic, Gram-negative, non-motile, dinitrogen-fixing rods that possess a soluble methane monooxygenase and utilize the serine pathway for carbon assimilation. Strains T4T, TCh1 and TY1 are moderately acidophilic organisms capable of growth between pH 4.2 and 7.5 (optimum 5.5-6.0) and between 5 and 30 degrees C (optimum 15 degrees C). The major phospholipid fatty acid is 18:1omega7c. The DNA G+C content of strain T4T is 63.3 mol%. The three strains possess almost identical 16S rRNA gene sequences and are most closely related to two previously identified species of Methylocella, Methylocella palustris (97% similarity) and Methylocella silvestris (97.5% similarity). DNA-DNA hybridization values of strain T4T with Methylocella palustris KT and Methylocella silvestris BL2T were respectively 27 and 36%. Thus, the tundra strains represent a novel species, for which the name Methylocella tundrae sp. nov. is proposed. Strain T4T (=DSM 15673T=NCIMB 13949T) is the type strain. PMID- 14742474 TI - Hongiella mannitolivorans gen. nov., sp. nov., Hongiella halophila sp. nov. and Hongiella ornithinivorans sp. nov., isolated from tidal flat sediment. AB - Three marine strains of the Cytophaga-Flavobacterium-Bacteroides group, designated JC2050T, JC2051T and JC2052T, were obtained from a single sediment sample of getbol, the Korean tidal flat. Comparative 16S rDNA sequence studies revealed that the test strains were not closely related to any validly published genera and that these strains were only distantly related to the genus Cyclobacterium (88.7-91.2%). Phylogenetic analyses demonstrated that the three getbol isolates formed a distinct monophyletic clade within the family Cytophagaceae. Physiological, biochemical and chemotaxonomic data also indicated that these three getbol isolates differed significantly from members of other genera and were sufficiently different from each other to be recognized as separate species. On the basis of polyphasic evidence, a new genus, Hongiella gen. nov., is proposed, with three novel species, Hongiella mannitolivorans sp. nov. (type strain JC2050T=IMSNU 14012T=DSM 15301T), Hongiella halophila sp. nov. (type strain JC2051T=IMSNU 14013T=DSM 15292T) and Hongiella ornithinivorans sp. nov. (type strain JC2052T=IMSNU 14014T=DSM 15282T). Hongiella mannitolivorans is the type species of the genus. PMID- 14742475 TI - Marinospirillum insulare sp. nov., a novel halophilic helical bacterium isolated from kusaya gravy. AB - A novel species that belongs to the genus Marinospirillum is described on the basis of phenotypic characteristics, phylogenetic analysis of 16S rRNA and gyrB gene sequences and DNA-DNA hybridization. Four strains of helical, halophilic, Gram-negative, heterotrophic bacteria were isolated from kusaya gravy, which is fermented brine that is used for the production of traditional dried fish in the Izu Islands of Japan. All of the new isolates were motile by means of bipolar tuft flagella, of small cell size, coccoid-body-forming and aerophilic; it was concluded that they belong to the same bacterial species, based on DNA-DNA hybridization values (>70% DNA relatedness). DNA G+C contents of the new strains were 42-43 mol% and they had isoprenoid quinone Q-8 as the major component. Phylogenetic analysis of 16S rRNA gene sequences indicated that the new isolates were members of the genus Marinospirillum; sequence similarity of the new isolates to Marinospirillum minutulum, Marinospirillum megaterium and Marinospirillum alkaliphilum was 98.5, 98.2 and 95.2%, respectively. Phylogenetic analysis based on the gyrB gene indicated that the new isolates had enough phylogenetic distance from M. minutulum and M. megaterium to be regarded as different species, with 84.7 and 78.7% sequence similarity, respectively. DNA-DNA hybridization showed that the new isolates had <36% DNA relatedness to M. minutulum and M. megaterium, supporting the phylogenetic conclusion. Thus, a novel species is proposed: Marinospirillum insulare sp. nov. (type strain, KT=LMG 21802T=NBRC 100033T). PMID- 14742476 TI - Petrotoga mexicana sp. nov., a novel thermophilic, anaerobic and xylanolytic bacterium isolated from an oil-producing well in the Gulf of Mexico. AB - A novel anaerobic, thermophilic, xylanolytic, motile rod-shaped bacterium with a sheath-like outer structure (toga) was isolated from a Mexican oil well in the Gulf of Mexico. Strain MET12T was a Gram-negative bacterium, reducing elemental sulfur, thiosulfate and sulfite to hydrogen sulfide. Its optimum growth conditions were 55 degrees C, pH 6.6, 3% NaCl and 0.15% MgCl2.6H2O. The DNA G+C content was 36.1 mol%. Phylogenetically, strain MET12T was related to members of genus Petrotoga, with similarities to Petrotoga mobilis, Petrotoga sibirica, Petrotoga miotherma and Petrotoga olearia varying from 97.6 to 98.8%. However DNA DNA relatedness values between these species and strain MET12T were lower than 70%. As strain MET12T (=DSM 14811T=CIP 107371T) was genomically and phenotypically different from existing Petrotoga species, it is proposed as the type strain of a novel species, Petrotoga mexicana sp. nov. PMID- 14742477 TI - Thermovibrio ammonificans sp. nov., a thermophilic, chemolithotrophic, nitrate ammonifying bacterium from deep-sea hydrothermal vents. AB - A thermophilic, anaerobic, chemolithoautotrophic bacterium was isolated from the walls of an active deep-sea hydrothermal vent chimney on the East Pacific Rise at 9 degrees 50' N. Cells of the organism were Gram-negative, motile rods that were about 1.0 microm in length and 0.6 microm in width. Growth occurred between 60 and 80 degrees C (optimum at 75 degrees C), 0.5 and 4.5% (w/v) NaCl (optimum at 2%) and pH 5 and 7 (optimum at 5.5). Generation time under optimal conditions was 1.57 h. Growth occurred under chemolithoautotrophic conditions in the presence of H2 and CO2, with nitrate or sulfur as the electron acceptor and with concomitant formation of ammonium or hydrogen sulfide, respectively. Thiosulfate, sulfite and oxygen were not used as electron acceptors. Acetate, formate, lactate and yeast extract inhibited growth. No chemoorganoheterotrophic growth was observed on peptone, tryptone or Casamino acids. The genomic DNA G+C content was 54.6 mol%. Phylogenetic analyses of the 16S rRNA gene sequence indicated that the organism was a member of the domain Bacteria and formed a deep branch within the phylum Aquificae, with Thermovibrio ruber as its closest relative (94.4% sequence similarity). On the basis of phylogenetic, physiological and genetic considerations, it is proposed that the organism represents a novel species within the newly described genus Thermovibrio. The type strain is Thermovibrio ammonificans HB-1T (=DSM 15698T=JCM 12110T). PMID- 14742478 TI - Methanocalculus chunghsingensis sp. nov., isolated from an estuary and a marine fishpond in Taiwan. AB - Three novel halotolerant, hydrogenotrophic methanogens, designated strains K1F9705bT, K1F9705c and O1F9704a, were isolated from an estuary in Eriln Shi, Taiwan, and from a nearby marine water aquaculture fishpond. These isolates were irregular cocci that stained Gram-negative. Strains K1F9705bT and K1F9705c were non-motile, but strain O1F9704a was weakly motile with flagella. They were able to use formate and H2/CO2 to form methane, but they could not catabolize acetate, methanol, trimethylamine or secondary alcohols. Acetate was required for cell growth. Tungsten greatly stimulated the growth of strains K1F9705bT and K1F9705c, but did not affect the growth of strain O1F9704a. Optimal pH and temperature for growth of these three isolates were respectively 7.2 and 37 degrees C. Optimal NaCl concentration for growth was 0.5% for strain O1F9704a and 1.0% for strains K1F9705c and K1F9705bT. Moreover, all strains grew well at up to 8-12% NaCl. Analysis of the 16S rRNA gene revealed that these isolates are members of the genus Methanocalculus, but are distinct from Methanocalculus taiwanensis, Methanocalculus pumilus and Methanocalculus halotolerans, with sequence similarities of 98.4, 98.3 and 98.2%, respectively. In addition, strain K1F9705bT possessed 85, 80, 37, 29 and 10% DNA-DNA relatedness to strain K1F9705c, strain O1F9704a, M. pumilus, M. halotolerans and M. taiwanensis, respectively. Analysis of protein profiles and the Mr of surface (S)-layer glycoprotein subunits showed that these three new isolates are closely related to, but distinct from, known Methanocalculus species. A novel species, Methanocalculus chunghsingensis sp. nov., is proposed for strains K1F9705bT, K1F9705c and O1F9704a. The type strain is K1F9705bT (=OCM 772T=DSM 14646T). PMID- 14742479 TI - Williamsia maris sp. nov., a novel actinomycete isolated from the Sea of Japan. AB - The taxonomic position of a marine actinomycete, strain SJS0289/JS1T, was determined using a polyphasic taxonomic approach. The organism, which had phenotypic properties consistent with its classification in the genus Williamsia, formed a distinct clade in the 16S rRNA gene tree together with the type strain of Williamsia muralis, but was readily distinguished from this species using DNA DNA relatedness and phenotypic data. The genotypic and phenotypic data show that the organism merits recognition as a new species of Williamsia. The name proposed for the new species is Williamsia maris; the type strain is SJS0289/JS1T (=DSM 44693T=JCM 12070T=KCTC 9945T=NCIMB 13945T). PMID- 14742480 TI - Bacillus odysseyi sp. nov., a round-spore-forming bacillus isolated from the Mars Odyssey spacecraft. AB - A round-spore-forming Bacillus species that produces an exosporium was isolated from the surface of the Mars Odyssey spacecraft. This novel species has been characterized on the basis of phenotypic traits, 16S rDNA sequence analysis and DNA-DNA hybridization. According to the results of these analyses, this strain belongs to the genus Bacillus and is a Gram-positive, aerobic, rod-shaped, endospore-forming eubacterium. Ultrathin sections of the spores showed the presence of an exosporium, spore coat, cortex and core. 16S rDNA sequence similarities between this strain, Bacillus fusiformis and Bacillus silvestris were approximately 96% and DNA-DNA reassociation values with these two bacilli were 23 and 17%, respectively. Spores of the novel species were resistant to desiccation, H2O2 and UV and gamma radiation. Of all strains tested, the spores of this strain were the most consistently resistant and survived all of the challenges posed, i.e. exposure to conditions of desiccation (100% survival), H2O2 (26% survival), UV radiation (10% survival at 660 J m(-2)) and gamma radiation (0.4% survival). The name proposed for this novel bacterium is Bacillus odysseyi sp. nov.; the type strain is 34hs-1T (=ATCC PTA-4993T=NRRL B-30641T=NBRC 100172T). PMID- 14742481 TI - Phylogenetic analysis of Streptomyces spp. isolated from potato scab lesions in Korea on the basis of 16S rRNA gene and 16S-23S rDNA internally transcribed spacer sequences. AB - The 16S rRNA gene sequences for 34 strains, including 11 isolates, were determined to classify scab-causing Streptomyces spp. and relatives isolated from potato scab lesions collected in Jeju, Korea. The 16S-23S rDNA internally transcribed spacer (ITS) sequences were determined to investigate whether the 16S 23S ITS region is useful for analysing intra- and interspecific relationships in these bacteria. On the basis of phylogenetic analysis of 16S rRNA gene sequences, most of the isolates were classified as Streptomyces scabiei and Streptomyces acidiscabies. Isolate KJO61 was placed in an ambiguous taxonomic position between Streptomyces reticuliscabiei and Streptomyces turgidiscabies. 16S-23S ITS region sequence analysis showed that tRNA genes were not found in this region of Streptomyces spp. The 16S-23S ITS regions of Streptomyces spp. exhibited various lengths and highly variable sequence similarities (35-100%) within strains as well as intra- and interspecies. It was revealed that Streptomyces europaeiscabiei could be clearly differentiated from Streptomyces scabiei. However, it was clarified that ITS regions are not useful in phylogenetic analysis of Streptomyces spp. PMID- 14742482 TI - Taxonomic study of neutrotolerant acidophilic actinomycetes isolated from soil and description of Streptomyces yeochonensis sp. nov. AB - Acidophilic actinomycete strains that represent the two major neutrotolerant clusters defined by numerical taxonomy (Seong, 1992) were the subject of a polyphasic taxonomic study. The centrotypes of each cluster, designated as strain JL164 (=KCTC 9924) of cluster 21 and strain CN732T (=KCTC 9926T=IMSNU 50114T=NRRL B-24245T) of cluster 13, were assigned initially to the genus Streptomyces on the basis of morphological and chemotaxonomic characteristics; this assignation was confirmed by 16S rRNA gene sequence data. Strain CN732T formed a distinct phyletic line within the Streptomyces tree, whereas strain JL164 was related closely to the type strain of Streptomyces mirabilis. It is evident from the present and previous studies that neutrotolerant acidophilic actinomycetes comprise taxonomically diverse groups within the variation encompassed by the genus Streptomyces. It is also apparent that strain CN732T and other members of cluster 13 merit recognition as a novel species, for which the name Streptomyces yeochonensis sp. nov. is proposed. PMID- 14742484 TI - Erysipelothrix inopinata sp. nov., isolated in the course of sterile filtration of vegetable peptone broth, and description of Erysipelotrichaceae fam. nov. AB - Based on 16S rRNA gene sequence comparison, an isolate that was detected in sterile-filtered vegetable broth was classified as a novel member of the Erysipelothrix line of descent of the Firmicutes. Strain MF-EP02T resembles members of the two species of Erysipelothrix with validly published names, Erysipelothrix rhusiopathiae and Erysipelothrix tonsillarum, in morphology, fatty acid composition, lack of menaquinones in aerobically and anaerobically grown cultures, DNA G+C content and peptidoglycan amino acid composition. Distinct differences in physiological characteristics, however, support the allocation of this isolate to a novel species of the genus Erysipelothrix, for which the name Erysipelothrix inopinata sp. nov. (type strain, MF-EP02T=DSM 15511T=CIP 107935T) is proposed. Members of the Erysipelothrix line of descent are included in the family Erysipelotrichaceae fam. nov. PMID- 14742483 TI - Bartonella chomelii sp. nov., isolated from French domestic cattle (Bos taurus). AB - Two strains of bacteria isolated from the blood of French domestic cows were found to be similar to Bartonella species on the basis of phenotypic characteristics. Genotypic analysis based on sequence comparison of the 16S rRNA and citrate synthase (gltA) genes and on DNA-DNA hybridization showed that the two isolates represent a distinct and new species of Bartonella. Moreover, the phylogenetic analysis inferred from comparison of 16S rRNA and gltA sequences demonstrated that the new Bartonella species is related to other ruminant-derived Bartonella species. The name Bartonella chomelii is proposed for the new species. The type strain of Bartonella chomelii sp. nov. is A828T (=CIP 107869T=CCUG47497T). PMID- 14742485 TI - Thermodesulfatator indicus gen. nov., sp. nov., a novel thermophilic chemolithoautotrophic sulfate-reducing bacterium isolated from the Central Indian Ridge. AB - A thermophilic, marine, anaerobic, chemolithoautotrophic, sulfate-reducing bacterium, strain CIR29812T, was isolated from a deep-sea hydrothermal vent site at the Kairei vent field on the Central Indian Ridge. Cells were Gram-negative motile rods that did not form spores. The temperature range for growth was 55-80 degrees C, with an optimum at 70 degrees C. The NaCl concentration range for growth was 10-35 g l(-1), with an optimum at 25 g l(-1). The pH range for growth was 6-6.7, with an optimum at approximately pH 6.25. H2 and CO2 were the only electron donor and carbon source found to support growth of the strain. However, several organic compounds were stimulatory for growth. Sulfate was used as electron acceptor, whereas elemental sulfur, thiosulfate, sulfite, cystine, nitrate and fumarate were not. No fermentative growth was observed with malate, pyruvate or lactate. The phenotypic characteristics of strain CIR29812T were similar to those of Thermodesulfobacterium hydrogeniphilum, a recently described thermophilic, chemolithoautotrophic sulfate-reducer. However, phylogenetic analyses of the 16S rRNA gene sequences showed that the new isolate was distantly related to members of the family Thermodesulfobacteriaceae (similarity values of less than 90%). The chemotaxonomic data (fatty acids and polar lipids composition) also indicated that strain CIR29812T could be distinguished from Thermodesulfobacterium commune, the type species of the type genus of the family Thermodesulfobacteriaceae. Finally, the G+C content of the genomic DNA of strain CIR29812T (46.0 mol%) was not in the range of values obtained for members of this family. On the basis of phenotypic, chemotaxonomic and genomic features, it is proposed that strain CIR29812T represents a novel species of a new genus, Thermodesulfatator, of which Thermodesulfatator indicus is the type species. The type strain is CIR29812T (=DSM 15286T=JCM 11887T). PMID- 14742486 TI - Amycolatopsis decaplanina sp. nov., a novel member of the genus with unusual morphology. AB - Strain DSM 44594T, which produces the glycopeptide antibiotic decaplanin, is a member of the genus Amycolatopsis based on 16S rRNA gene sequence analysis and chemotaxonomic properties. It is the first member of this genus that is reported to form pseudosporangia, which resemble those of members of the genus Kibdelosporangium. Phylogenetically, the novel taxon is related to Amycolatopsis orientalis, Amycolatopsis lurida, Amycolatopsis azurea, Amycolatopsis japonica and Amycolatopsis keratiniphila. Morphological, cultural and physiological properties, the production of a unique glycolipid and DNA-DNA similarity of <55% with phylogenetically related strains reveal that strain DSM 44594T represents a novel species of the genus, for which the name Amycolatopsis decaplanina sp. nov. (type strain, FH 1845T=DSM 44594T=NRRL B-24209T) is proposed. PMID- 14742487 TI - Hespellia stercorisuis gen. nov., sp. nov. and Hespellia porcina sp. nov., isolated from swine manure storage pits. AB - Four Gram-positive-staining, strictly anaerobic, non-spore-forming, rod-shaped organisms were isolated from a pig manure storage pit. Comparative 16S rRNA gene sequence analysis revealed that the isolates belonged to two related but distinct groups. Sequence analysis showed that the two groups of isolates were highly related to each other (approx. 97% 16S rRNA gene sequence similarity), forming a distinct cluster within the Clostridium coccoides suprageneric rDNA grouping. Biochemical and physiological studies confirmed the division of the isolates into two related, albeit distinct, groups. Based on both phenotypic and phylogenetic evidence, it is proposed that the unidentified rod-shaped isolates from pig manure should be classified in a novel genus, Hespellia gen. nov., as Hespellia stercorisuis sp. nov. and Hespellia porcina sp. nov. The type species of the novel genus is H. stercorisuis (type strain, PC18T=NRRL B-23456T=CCUG 46279T=ATCC BAA-677T) and the type strain of H. porcina is PC80T (=NRRL B-23458T=ATCC BAA 674T). PMID- 14742488 TI - Nocardiopsis alkaliphila sp. nov., a novel alkaliphilic actinomycete isolated from desert soil in Egypt. AB - An alkaliphilic actinomycete strain, designated YIM 80379T, was isolated from a soil sample collected from the eastern desert of Egypt and subjected to polyphasic taxonomy. The strain produced substrate and aerial mycelia on different media, with an optimum pH for growth of 9.5-10 and scarce or no growth at pH 7. Strain YIM 80379T contained meso-diaminopimelic acid, no diagnostic sugars, type PIII phospholipids and MK-10(H6) and MK-10(H8) as the predominant menaquinones. All of these characters assign isolate YIM 80379T consistently to the genus Nocardiopsis. This was confirmed by 16S rDNA analysis. It can be differentiated from all Nocardiopsis species with validly published names by phenotypic characteristics, phylogenetic analysis and DNA-DNA hybridization results. On the basis of polyphasic evidence, a novel species, Nocardiopsis alkaliphila sp. nov., is proposed. The type strain of the species is YIM 80379T (=CCTCC AA001031T=DSM 44657T). PMID- 14742489 TI - Blastococcus saxobsidens sp. nov., and emended descriptions of the genus Blastococcus Ahrens and Moll 1970 and Blastococcus aggregatus Ahrens and Moll 1970. AB - Ten Gram-positive, aerobic bacterial strains with coccoid cells occurring singly, in pairs, tetrads and small aggregates were isolated from the surface of marble and calcareous stones. The peptidoglycan contained meso-diaminopimelic acid as the diagnostic diamino acid. The major menaquinone was MK-9(H4). The cellular fatty acid pattern consisted mainly of iso-branched chain components. According to their phylogenetic position, the organisms are members of the family Geodermatophilaceae, clustering close to Blastococcus aggregatus DSM 4725T. Based on a combination of phenotypic, chemotaxonomic and genotypic characteristics, strain BC444T and seven relatives group apart from strains BC412 (=DSM 44517) and BC521 (=DSM 44518), which themselves form two individual lineages. DNA-DNA reassociation studies revealed that members of the three lineages were only remotely related to B. aggregatus DSM 4725T; as the strain-rich cluster containing BC444T shared low DNA similarity values with strains BC412 and BC521, it will be described as a novel species of Blastococcus, for which the name Blastococcus saxobsidens sp. nov. is proposed. The type strain is BC444T (=DSM 44509T=NRRL 24246T). PMID- 14742490 TI - Vibrio hispanicus sp. nov., isolated from Artemia sp. and sea water in Spain. AB - Three Gram-negative, small, motile, rod-shaped bacteria were isolated from Artemia sp. and sea water in Barcelona, Spain, during 1990 and 1991. They were fermentative, oxidase-positive, sensitive to vibriostatic agent O/129, arginine dihydrolase-positive, lysine and ornithine decarboxylase-negative and grew in the absence of NaCl. They differed from phenotypically related species by their ability to grow at 4 degrees C and utilize L-rhamnose. Cloning of the 16S rRNA gene of the type strain produced two different 16S rRNA gene sequences, which differed by 15 bases (0.99%); comparison of these sequences with those deposited in GenBank showed close relationships with Vibrio proteolyticus (97.6% similarity), Vibrio diazotrophicus (97.9%), Vibrio campbellii (96.8%) and Vibrio alginolyticus (96.8%), among others. DNA-DNA hybridization levels with the closest phylogenetically related Vibrio species were <26.4%. Sufficient evidence is provided to support the identity of the three strains analysed as members of a novel species of the genus Vibrio, for which the name Vibrio hispanicus sp. nov. is proposed, with the type strain LMG 13240T (=CAIM 525T=VIB 213T). PMID- 14742491 TI - Reclassification of Amycolatopsis orientalis subsp. lurida Lechevalier et al. 1986 as Amycolatopsis lurida sp. nov., comb. nov. AB - Amycolatopsis orientalis subsp. lurida DSM 43134T differs significantly from the type strain of A. orientalis, A. orientalis subsp. orientalis DSM 40040T, and from other species of the genus in genomic and metabolic properties. Its elevation to species status as Amycolatopsis lurida sp. nov., comb. nov. is justified. PMID- 14742492 TI - Nitratireductor aquibiodomus gen. nov., sp. nov., a novel alpha-proteobacterium from the marine denitrification system of the Montreal Biodome (Canada). AB - The Montreal Biodome operates a methanol-fed denitrification system that treats the water in its three million litre marine mesocosm. An unknown bacterium, named strain NL21T, was isolated from this system on TSA and R2A agar. The organism is a Gram-negative, rod-shaped (1x3 microm) facultative aerobe. Optimal growth conditions on R2A agar are 30-35 degrees C, pH 7-7.5 and 1% (w/w) NaCl. Phylogenetic analysis of the 16S rDNA sequence reveals that strain NL21T forms a novel lineage in the family 'Phyllobacteriaceae' within the alpha2 subgroup of the Proteobacteria. The closest related genera are Aminobacter, Pseudaminobacter, Mesorhizobium and Defluvibacter. Major cellular fatty acids are C(18:1)omega7c (75%), C(19:0)omega8c cyclopropane (9.4%) and C(18:0) (4.2%). The DNA G+C content of strain NL21T (57 mol%) differs from those of all other described members of the 'Phyllobacteriaceae' (60-64 mol%). Strain NL21T reduces nitrate to nitrite, but does not reduce nitrite to nitrogen gas. Only a few sugars and amino acids can serve as carbon sources. Strain NL21T is able to grow without salt and tolerates up to 5% NaCl. Phylogenetic analysis, as well as physiological and biochemical tests, showed that strain NL21T was different from all other members of the 'Phyllobacteriaceae' with validly published names. Strain NL21T therefore represents a novel genus, for which the name Nitratireductor aquibiodomus gen. nov., sp. nov. is proposed, with the type strain NL21T (=DSM 15645T=ATCC BAA 762T). PMID- 14742493 TI - A novel psychrotolerant member of the hymenomycetous yeasts from Antarctica: Cryptococcus watticus sp. nov. AB - Soil, snow and organic material, collected in November 1997 from the Vestvold Hills, Davis Base, Antarctica, were screened for yeasts. Two isolates, which were shown to be indistinguishable by rDNA sequencing and protein analysis by SDS PAGE, are described in this communication as a novel species, Cryptococcus watticus sp. nov. (type culture, CBS 9496T=NRRL Y-27556T). Sequence analyses of the 26S rDNA D1/D2 region placed C. watticus in the hymenomycetous yeasts in a cluster with Holtermannia corniformis and Cryptococcus nyarrowii. This species has been allocated to the genus Cryptococcus on the basis of physiological and morphological characteristics. PMID- 14742494 TI - Should names reflect the evolution of bacterial species? AB - Systematics is the process of characterizing and arranging bacterial diversity in an orderly manner, recognizing groups of similar organisms in a hierarchical scheme whose basic entity is the species. To allow the exchange of scientific knowledge, taxa have to be named. Taxa are not static entities since they are subject to evolution, the direction of which can be inferred by using a wide range of techniques targeting specific traits. However, it is not clear how this dynamism should be reflected in taxonomic nomenclature. In the present report, several considerations are presented that deal with the relationship between the evolution of taxa and their nomenclature; an example is given which concerns the nomenclature of the species Lactobacillus delbrueckii. PMID- 14742499 TI - Status of strains that contravene Rules 27(3) and 30 of the Bacteriological Code. Request for an opinion. AB - In the period from January 2001, at least 207 new names proposed in articles in the International Journal of Systematic and Evolutionary Microbiology or cited in Validation Lists are not in accordance with Rules 27(3) and 30 of the Bacteriological Code. The purpose of the present Request for an Opinion is to clarify the status of the names listed and to provide a solution whereby they may be considered to be validly published. PMID- 14742500 TI - Reconsideration of Arthrobacter ilicis (Mandel et al. 1961) Collins et al. 1982 as a plant-pathogenic species. Proposal to emend the authority and description of the species. Request for an opinion. AB - Strains now considered to represent the type strain of Arthrobacter ilicis, described as a pathogen of American holly, are not identical. The designated type strain does not represent this pathogen. However, one of the other strains sourced to the type strain of the pathogen does appear to be authentic, but is not a member of A. ilicis. It is proposed that A. ilicis is an unrelated species, not a pathogen of American holly. The nomenclature of A. ilicis can be rectified by emending the authority and by emending the species description to recognize this species as a novel species that is not a plant pathogen. The pathogen of American holly then becomes a novel pathovar, Curtobacterium flaccumfaciens pv. ilicis. The opinion of the Judicial Commission is sought. PMID- 14742501 TI - Contribution of CsrR-regulated virulence factors to the progress and outcome of murine skin infections by Streptococcus pyogenes. AB - Streptococcus pyogenes with null mutations in the csrRS regulatory locus are highly virulent in mice due to derepression of hyaluronic acid capsule synthesis and exotoxins, e.g., streptolysin S (SLS) and pyrogenic exotoxin B (SpeB). We generated derivatives of a DeltacsrRS strain that also carry deletions in hasAB (leading to an acapsular phenotype) or in sagA (phenotypically SLS-) or an interruption of speB (SpeB-) to test the relative contributions of these factors to the development of necrotic skin lesions. Inoculation of 2 x 10(6) to 4 x 10(6) CFU of either acapsular or SLS- strains into hairless mice resulted in lesions approximately 70% smaller than those of the DeltacsrRS parent strain. Elimination of SLS also reduced lethality from 100% to 0% at this inoculum (P < 10(-7); Fisher exact test). In contrast, SLS+ SpeB- mutants yielded lesions that were only 41% smaller than the parent strain (t = 2.2; P = 0.04), but only 3 the 17 lesions had dermal sloughing (P = 10(-5)). The nonulcerative lesions associated with SpeB- strains appeared pale with surrounding erythema. We conclude that capsule and SLS contribute to the subcutaneous spread of S. pyogenes and to a fatal outcome of infection. SpeB facilitates early dermal ulceration but has minor influence on lesion size and mortality. Large ulcerative lesions are observed only when both toxins are present. PMID- 14742502 TI - A K+ yptake protein, TrkA, is required for serum, protamine, and polymyxin B resistance in Vibrio vulnificus. AB - Vibrio vulnificus, a highly virulent marine bacterium, is the causative agent of both serious wound infections and fatal septicemia in many areas of the world. To identify the genes required for resistance to human serum, we constructed a library of transposon mutants of V. vulnificus and screened them for hypersensitivity to human serum. Here we report that one of the isolated serum susceptible mutants had a mutation in an open reading frame identified as trkA, a gene encoding an amino acid sequence showing high identity to that of TrkA of Vibrio alginolyticus, a protein required for the uptake of potassium. A trkA isogenic mutant was constructed via insertional inactivation, and it was significantly more easily killed by human serum, protamine, or polymyxin B than was the wild type. At K+ concentrations of 1 to 20 mM, this isogenic mutant showed attenuated growth compared to the wild-type strain. In addition, infection experiments demonstrated virulence attenuation when this mutant was administered intraperitoneally or subcutaneously to both normal and iron-treated mice, indicating that TrkA may modulate the transport of potassium and resistance to host innate defenses and that it is important for virulence in mice. PMID- 14742503 TI - Role of B7 costimulatory molecules in immune responses and T-helper cell differentiation in response to recombinant HagB from Porphyromonas gingivalis. AB - In addition to antigen-specific signals mediated through the T-cell receptor, T cells also require antigen nonspecific costimulation for activation. The B7 family of molecules on antigen-presenting cells, which include B7-1 (CD80) and B7 2 (CD86), play important roles in providing costimulatory signals required for development of antigen-specific immune responses. Hemagglutinin B (HagB) is a nonfimbrial adhesin of the periodontopathic microorganism Porphyromonas gingivalis and is thought to be involved in the attachment of the bacterium to host tissues. However, the immune mechanisms involved in responses to HagB and their roles in pathogenesis have yet to be elucidated. Therefore, the purpose of this study was to determine the role of B7 costimulatory molecules on T-helper cell differentiation for the induction of immune responses to HagB. Mice deficient in either or both of the costimulatory molecules B7-1 and B7-2 were used to explore their role in immune responses to HagB after subcutaneous immunization. B7-1(-/-) mice had levels of immunoglobulin G (IgG) anti-HagB antibody activity in serum similar to those of wild-type mice, whereas lower serum IgG anti-HagB antibody responses were seen in B7-2(-/-) mice. Moreover, significantly lower numbers of IgG antibody-secreting cells and lower levels of CD4(+)-T-cell proliferation were observed in B7-2(-/-) mice compared to wild-type mice. No serum IgG response to HagB was detected in B7-1/B7-2(-/-) mice. Analysis of the subclass of the serum IgG responses and the cytokines induced in response to HagB revealed that B7-2(-/-) mice had significantly lower IgG1 and higher IgG2a anti-HagB antibody responses compared to wild-type mice. The B7-2(-/-) mice also had significantly reduced levels of interleukin-4 (IL-4) and IL-5 and enhanced level of gamma interferon. Furthermore, assessment of B7-1 and B7-2 expression on B cells and macrophages derived from wild-type BALB/c mice after in vitro stimulation with HagB revealed a predominant upregulation in the expression of the B7-2 costimulatory molecule on B cells and macrophages. Essentially no change was seen in the expression of B7-1. Taken together, these results suggest a critical role for B7, especially B7-2, for the preferential induction of a Th2 like response to HagB. PMID- 14742504 TI - Surfactant protein A modulates the inflammatory response in macrophages during tuberculosis. AB - Tuberculosis leads to immune activation and increased human immunodeficiency virus type 1 (HIV-1) replication in the lung. However, in vitro models of mycobacterial infection of human macrophages do not fully reproduce these in vivo observations, suggesting that there are additional host factors. Surfactant protein A (SP-A) is an important mediator of innate immunity in the lung. SP-A levels were assayed in the human lung by using bronchoalveolar lavage (BAL). There was a threefold reduction in SP-A levels during tuberculosis only in the radiographically involved lung segments, and the levels returned to normal after 1 month of treatment. The SP-A levels were inversely correlated with the percentage of neutrophils in BAL fluid, suggesting that low SP-A levels were associated with increased inflammation in the lung. Differentiated THP-1 macrophages were used to test the effect of decreasing SP-A levels on immune function. In the absence of infection with Mycobacterium tuberculosis, SP-A at doses ranging from 5 to 0.01 micro g/ml inhibited both interleukin-6 (IL-6) production and HIV-1 long terminal repeat (LTR) activity. In macrophages infected with M. tuberculosis, SP-A augmented both IL-6 production and HIV-1 LTR activity. To better understand the effect of SP-A, we measured expression of CAAT/enhancer binding protein beta (C/EBPbeta), a transcription factor central to the regulation of IL-6 and the HIV-1 LTR. In macrophages infected with M. tuberculosis, SP-A reduced expression of a dominant negative isoform of C/EBPbeta. These data suggest that SP-A has pleiotropic effects even at the low concentrations found in tuberculosis patients. This protein augments inflammation in the presence of infection and inhibits inflammation in uninfected macrophages, protecting uninvolved lung segments from the deleterious effects of inflammation. PMID- 14742505 TI - Porphyromonas gingivalis genes involved in fimA regulation. AB - Porphyromonas gingivalis is an important component of the complex plaque biofilm that is a direct precursor of periodontal disease. The major fimbriae are required for attachment to oral surfaces and are an important virulence factor. Fimbrillin (FimA) expression in P. gingivalis is inhibited by surface molecule of Streptococcus cristatus, an early colonizer of dental plaque. In this study, differential display PCR was used to identify P. gingivalis genes that are regulated in response to S. cristatus. Of several differentially expressed genes, pg2131 and pg2167 were upregulated by S. cristatus signaling molecules. A null mutant of pg2167 did not transcriptionally regulate fimA following exposure to S. cristatus. In fact, fimA transcription was enhanced in the pg2167 mutant, suggesting that pg2167 may act to repress fimA expression. In contrast, a mutation in pg2131 did not affect transcription of fimA in the presence of S. cristatus. However, production of fimbrillin was significantly diminished in the pg2131 mutant, implicating involvement in posttranscriptional regulation in fimbriation. These data suggest that P. gingivalis fimbriation is controlled by more than one regulation mechanism, involving both transcriptional and posttranscriptional processes. PMID- 14742506 TI - Sequence analysis of p44 homologs expressed by Anaplasma phagocytophilum in infected ticks feeding on naive hosts and in mice infected by tick attachment. AB - The 44-kDa immunodominant outer membrane proteins (P44 proteins) of Anaplasma phagocytophilum are encoded by the p44 polymorphic multigene family. The present study examined p44 expression and analyzed the cDNA sequences of various p44 transcripts from the spleens and blood of mice infected by the bites of ticks infected with the A. phagocytophilum NTN-1 strain or of naturally infected nymphal ticks and in the salivary glands and midgut tissues of these ticks. A total of 300 p44 cDNAs were subjected to sequence analysis. Of these, 40 distinct p44 species were found, and all of these had orthologs in the A. phagocytophilum HZ strain genome that shared 95 to 100% base sequence identity. The number of unique p44 species expressed in mouse blood was greater than that for mouse spleens. Higher numbers of different p44 transcripts were also expressed in the salivary glands of ticks than in the midgut tissues. Variations in the sequences of the same p44 cDNA species within a single A. phagocytophilum strain and among different strains were concentrated in the conserved regions flanking the central hypervariable region of p44 genes. No mosaic sequences derived from two or more p44 species were found within the p44 hypervariable region. The conservation of the hypervariable region of each p44 cDNA species of A. phagocytophilum in naturally infected ticks and in different geographic isolates suggests that each A. phagocytophilum genome carries a set of p44 paralogs to be expressed. Thus, a large but restricted repertoire of p44 hypervariable sequences exists in A. phagocytophilum strains in the Northeastern United States. PMID- 14742507 TI - Release of a potent polymorphonuclear leukocyte chemoattractant is regulated by white-opaque switching in Candida albicans. AB - Previous studies employing transmembrane assays suggested that Candida albicans and related species, as well as Saccharomyces cerevisiae, release chemoattractants for human polymorphonuclear leukocytes (PMNs). Because transmembrane assays do not definitively distinguish between chemokinesis and chemotaxis, single-cell chemotaxis assays were used to confirm these findings and test whether mating-type or white-opaque switching affects the release of attractant. Our results demonstrate that C. albicans, C. dubliniensis, C. tropicalis, C. parapsilosis, and C. glabrata release bona fide chemoattractants for PMNs. S. cerevisiae, however, releases a chemokinetic factor but not a chemoattractant. Characterization of the C. albicans chemoattractant revealed that it is a peptide of approximately 1 kDa. Whereas the mating type of C. albicans did not affect the release of chemoattractant, switching did. White phase cells released chemoattractant, but opaque-phase cells did not. Since the opaque phase of C. albicans represents the mating-competent phenotype, it may be that opaque-phase cells selectively suppress the release of chemoattractant to facilitate mating. PMID- 14742508 TI - Expression of amoebapores is required for full expression of Entamoeba histolytica virulence in amebic liver abscess but is not necessary for the induction of inflammation or tissue damage in amebic colitis. AB - Entamoeba histolytica trophozoites produce amoebapores, a family of small amphipathic peptides capable of insertion into bacterial or eukaryotic membranes and causing cellular lysis. Recently, E. histolytica trophozoites that are totally deficient in the production of amoebapore-A were created through a gene silencing mechanism (R. Bracha, Y. Nuchamowitz, and D. Mirelman, Eukaryot. Cell 2:295-305, 2003). Here we tested the virulence of amoebapore A(-) trophozoites in models of the two major forms of amebic disease: amebic liver abscess and amebic colitis. We demonstrate that amoebapore expression is required for full virulence in the SCID mouse model of amebic liver abscess, but E. histolytica trophozoites that do not express amoebapore-A can still cause inflammation and tissue damage in infected human colonic xenografts. These data are consistent with the concept that tissue damage may proceed by different mechanisms in amebic liver abscess compared to amebic colitis. PMID- 14742509 TI - Patterns of variations in Escherichia coli strains that produce cytolethal distending toxin. AB - A collection of 20 Escherichia coli strains that produce cytolethal distending toxin (CDT) were analyzed for their virulence-associated genes. All of these strains were serotyped, and multiplex PCR analysis was used to ascertain the presence of genes encoding other virulence factors, including Shiga toxin, intimin, enterohemolysin, cytotoxic necrotizing factor type 1 (CNF1) and CNF2, heat-stable toxin, and heat-labile toxin. These CDT-producing strains possessed various combinations of known virulence genes, some of which have not been noted before. Partial cdtB sequences were obtained from 10 of these strains, and their predicted CdtB sequences were compared to known E. coli CdtB sequences; some of the sequences were identical to known CdtB sequences, but two were not. PCR primers based on sequence differences between the known cdt sequences were tested for their ability to detect CDT producers and to determine CDT type. Correlations between the type of CDT produced, the presence of other virulence properties, and overall strain relatedness revealed that the CDT producers studied here can be divided into three general groups, with distinct differences in CDT type and in their complement of virulence-associated genes. PMID- 14742510 TI - Host cell contact-induced transcription of the type IV fimbria gene cluster of Actinobacillus pleuropneumoniae. AB - Type IV pili (Tfp) of gram-negative species share many characteristics, including a common architecture and conserved biogenesis pathway. Much less is known about the regulation of Tfp expression in response to changing environmental conditions. We investigated the diversity of Tfp regulatory systems by searching for the molecular basis of the reported variable expression of the Tfp gene cluster of the pathogen Actinobacillus pleuropneumoniae. Despite the presence of an intact Tfp gene cluster consisting of four genes, apfABCD, no Tfp were formed under standard growth conditions. Sequence analysis of the predicted major subunit protein ApfA showed an atypical alanine residue at position -1 from the prepilin peptidase cleavage site in 42 strains. This alanine deviates from the consensus glycine at this position in Tfp from other species. Yet, cloning of the apfABCD genes under a constitutive promoter in A. pleuropneumoniae resulted in pilin and Tfp assembly. Tfp promoter-luxAB reporter gene fusions demonstrated that the Tfp promoter was intact but tightly regulated. Promoter activity varied with bacterial growth phase and was detected only when bacteria were grown in chemically defined medium. Infection experiments with cultured epithelial cells demonstrated that Tfp promoter activity was upregulated upon adherence of the pathogen to primary cultures of lung epithelial cells. Nonadherent bacteria in the culture supernatant exhibited virtually no promoter activity. A similar upregulation of Tfp promoter activity was observed in vivo during experimental infection of pigs. The host cell contact-induced and in vivo-upregulated Tfp promoter activity in A. pleuropneumoniae adds a new dimension to the diversity of Tfp regulation. PMID- 14742511 TI - Two TonB systems in Actinobacillus pleuropneumoniae: their roles in iron acquisition and virulence. AB - Iron acquisition in vivo by Actinobacillus pleuropneumoniae depends upon a functional TonB system. Tonpitak et al. (W. Tonpitak, S. Thiede, W. Oswald, N. Baltes, and G.-F. Gerlach, Infect. Immun. 68:1164-1170, 2000) have described one such system, associated with tbpBA encoding the transferrin receptor, and here we report a second, termed tonB2. This gene cluster (exbB2-exbD2-tonB2) is highly homologous to those in other Pasteurellaceae, unlike the earlier system described (now termed tonB1), suggesting that it is the indigenous system for this organism. Both tonB2 and tonB1 are upregulated upon iron restriction. TonB2, but not TonB1, was found to be essential for growth in vitro when the sole source of iron was hemin, porcine hemoglobin, or ferrichrome. In the case of iron provided as iron-loaded porcine transferrin, neither tonB mutant was viable. The tonB1 phenotype could be explained by a polar effect of the mutation on transcription of downstream tbp genes. We propose that TonB2 is crucial for the acquisition of iron provided in this form, interacting with accessory proteins of the TonB1 system that have been demonstrated to be necessary by Tonpitak et al. TonB2 appears to play a much more important role in A. pleuropneumoniae virulence than TonB1. In an acute porcine infection model, the tonB2 mutant was found to be highly attenuated, while the tonB1 mutant was not. We hypothesize that acquisition of the tonB1-tbp gene cluster confers a biological advantage through its capacity to utilize transferrin-iron but that TonB1 itself plays little or no part in this process. PMID- 14742512 TI - Binding and agglutination of Streptococcus pneumoniae by human surfactant protein D (SP-D) vary between strains, but SP-D fails to enhance killing by neutrophils. AB - Recombinant human surfactant protein D (SP-D) expressed in Escherichia coli, consisting of the head and neck regions of the native molecule, bound to all strains of Streptococcus pneumoniae that were tested, but the extent of binding varied between strains of differing capsular serotypes. The recombinant protein expressed in the yeast Pichia pastoris did not bind. Full-length native SP-D aggregated pneumococci in a calcium-dependent manner that was inhibited by maltose acting as a competitive sugar. The ability of SP-D to modulate the uptake and killing of pneumococci by human neutrophils was also addressed. Neither recombinant truncated SP-D nor native full-length SP-D enhanced the killing of pneumococci by human neutrophils. Aggregation of pneumococci varied not only between strains of the same multilocus sequence type and different serotypes but also between strains of the same serotype. However, use of recombinant strains in which the serotype had been changed showed that the degree of aggregation was influenced by the capsular type. Indeed, a 19F serotype strain which was not aggregated by SP-D did exhibit aggregation when the original isogenic strain was capsule switched to capsular serotype 3. However, although our results show that SP-D is capable of aggregating most pneumococci, no correlation between the degree of aggregation and the capsule or multilocus sequence type of the pneumococcus was clearly apparent. Therefore, although the capsule serotype is not the only determinant of aggregation by SP-D, the data presented here indicate that it does have a role to play. PMID- 14742513 TI - Disruption of putative regulatory loci in Listeria monocytogenes demonstrates a significant role for Fur and PerR in virulence. AB - The ability to adapt to adverse environmental conditions encountered in food and during host infection is a sine qua non for a successful Listeria monocytogenes infection. This ability is likely to depend on complex regulatory pathways controlled by a number of key regulators. We utilized the pORI19 plasmid integration system to analyze the role of six putative regulatory loci in growth under suboptimal environmental conditions and during murine infection. Disruption of loci encoding a topoisomerase III (lmo2756), a putative methyltransferase (lmo0581), and a regulator of the MarR family (lmo1618) revealed roles for the methyltransferase and the MarR regulator in growth under environmental stress conditions. However, plasmid integration into these loci had no impact on virulence potential in the murine model of infection. Disruption of the alternative sigma factor Sigma-H resulted in a mutant that demonstrated reduced growth potential in minimal medium. Murine studies indicated a minor role for this sigma factor in the infectious process. Strikingly, disruption of both perR and fur loci resulted in mutants that are significantly affected in virulence for mice, with the fur mutant demonstrating the greatest reduction in virulence potential. Both perR and fur mutants demonstrated increased resistance to hydrogen peroxide and the fur mutant was sensitive to low-iron conditions. The virulence defect of both fur and perR mutants could be rescued by iron-overload after esculetin treatment of mice, suggesting that the in vivo role of these gene products is to procure iron for bacterial growth. PMID- 14742514 TI - Cytokine production in whole blood cultures from a fishing community in an area of high endemicity for Schistosoma mansoni in Uganda: the differential effect of parasite worm and egg antigens. AB - The human host is continuously exposed to the egg and the adult worm developmental stages of Schistosoma mansoni during chronic infections with the parasite. To assess the cytokine responses induced by these different costimulating stages and how they are influenced by host age and infection intensity, whole blood samples from a cross-sectional cohort of 226 members of a Ugandan fishing community who had been resident in an area with high transmission of S. mansoni for the previous 10 years or from birth were stimulated with S. mansoni egg antigen (SEA) or worm antigen (SWA). SWA-specific gamma interferon (IFN-gamma) production increased with age, and the levels of SWA- and SEA specific interleukin 3 (IL-3) were weakly correlated with schistosome infection intensity. The production of most cytokines was little affected by age or infection intensity but was either SEA or SWA specific. One hundred thirty-two members of the cohort coproduced IL-5 and IL-13 specifically in response to SWA, whereas only 15 produced these cytokines, and at much lower levels, in response to SEA. IL-10, IL-4, and IFN-gamma were also produced in response to SWA, whereas the response to SEA consisted almost exclusively of IL-10. Our results suggest that, in contrast to what has been described for the murine model of S. mansoni and during acute human infections, chronic intense exposure to and infection with S. mansoni in this cohort resulted in very low levels of response to SEA in vitro in the presence of a vigorous and mixed Th1-Th2 response to SWA. PMID- 14742515 TI - Measurement of antibody levels against region II of the erythrocyte-binding antigen 175 of Plasmodium falciparum in an area of malaria holoendemicity in western Kenya. AB - Region II of the 175-kDa erythrocyte-binding antigen (EBA-175RII) of Plasmodium falciparum is functionally important in sialic acid-dependent erythrocyte invasion and is considered a prime target for an invasion-blocking vaccine. The objectives of this study were to (i) determine the prevalence of anti-EBA-175RII antibodies in a naturally exposed population, (ii) determine whether naturally acquired antibodies have a functional role by inhibiting binding of EBA-175RII to erythrocytes, and (iii) determine whether antibodies against EBA-175RII correlate with immunity to clinical malaria. We treated 301 lifelong residents of an area of malaria holoendemicity in western Kenya for malaria, monitored them during a high-transmission season, and identified 33 individuals who were asymptomatic despite parasitemia (clinically immune). We also identified 50 clinically susceptible individuals to serve as controls. These 83 individuals were treated and monitored again during the subsequent low-transmission season. Anti-EBA 175RII antibodies were present in 98.7% of the individuals studied. The antibody levels were relatively stable between the beginning and end of the high transmission season and correlated with the plasma EBA-175RII erythrocyte-binding inhibitory activity. There was no difference in anti-EBA-175RII levels or plasma EBA-175RII erythrocyte-binding-inhibitory activity between clinically immune and clinically susceptible groups. However, these parameters were higher in nonparasitemic than in parasitemic individuals at enrollment. These results suggest that although antibodies against EBA-175RII may be effective in suppressing some of the wild parasite strains, EBA-175RII is unlikely to be effective as a monovalent vaccine against malaria, perhaps due to allelic heterogeneity and/or presence of sialic acid-independent strains. PMID- 14742516 TI - Host-inducible immunogenic sphingomyelinase-like protein, Lk73.5, of Leptospira interrogans. AB - Leptospira interrogans causes a variety of clinical syndromes in animals and humans. Although much information has accumulated on the importance of leptospiral lipopolysaccharide in protective antibody responses, relatively little is known about proteins that participate in immune responses. Identification of those proteins induced only in the host is particularly difficult. Using a novel double-antibody screen designed to identify clones in a gene library of L. interrogans serovar Pomona expressing host-inducible proteins, we have characterized a gene (lk75.3) encoding a sphingomyelinase-like preprotein of 648 amino acids with cytotoxic activity for equine pulmonary endothelial cells and weak hemolytic activity for equine and rabbit erythrocytes. lk73.5 was found as a single gene copy in all serovars of L. interrogans but not in other Leptospira spp. except L. inadai. The open reading frame (ORF) for Lk73.5 is followed by another partially homologous sequence containing an ORF (sph-like 2) for a 28.7-kDa peptide. Lk73.5 and Sph-like 2 share 95.1 and 97.7% amino acid identity with putative sphingomyelinases Sph2 and Sph1 (N terminus) from L. interrogans serovar Lai (S.-X. Ren, G. Fu, X.-G. Jiangk, R. Zeng, Y.-G. Miao, H. Xu, Y.-X. Zhang, H. Xiong, G. Lu, L.-F. Lu, H.-Q. Jiang, J. Jia, Y.-F. Tu, J.-X. Jiang, W.-Y. Gu, Y.-Q. Zhang, Z. Cai, H.-H. Sheng, H.-F. Yin, Y. Zhang, G.-F. Zhu, M. Wank, H.-L. Huangk, Z. Qian, S.-Y. Wang, Wei Ma, Z.-J. Yao, Y. Shen, B. Q. Qiang, Q.-C. Xia, X.-K. Guo, A. Danchinq, I. S. Girons, R. L. Somerville, Y. M. Wen, M.-H. Shik, Z. Chen, J.-G. Xuk, and G.-P. Zhao, Nature 422:88-893, 2003). Substantial homologies to sphingomyelinases from other leptospiras and other bacteria are also present. Lk73.5 was not detected in leptospiras cultured at 30 or 37 degrees C. The recombinant protein reacted strongly with sera from recently infected mares but not with sera from horses vaccinated with commercial pentavalent bacterin. The host-inducible immunogenic Lk73.5 should have value in distinguishing vaccine from infection immune response. PMID- 14742517 TI - Immunogenicity against human papillomavirus type 16 virus-like particles is strongly enhanced by the PhoPc phenotype in Salmonella enterica serovar Typhimurium. AB - Recombinant Salmonella strains have been widely used to deliver heterologous antigens and induce immune responses in vaccinated animals and humans. It remains to be established, however, how these bacteria mount an immune response; this has prevented the rational design of vaccines. Here we report for the first time that a particular genetic program, PhoPc, is necessary for recombinant Salmonella strains to induce an antibody response to a heterologous antigen, the human papillomaviruses type 16 (HPV16) virus-like particle (VLP). The PhoPc phenotype results from a point mutation in phoQ, the gene encoding the sensor component of a two-component regulatory system (PhoP-PhoQ) that controls the expression of a number of virulence factors in Salmonellae. To demonstrate that immunogenicity of the viral antigen expressed by the bacterial vector was dependent on the PhoPc phenotype, we have expressed the phoQ mutant gene (phoQ24) in two differently attenuated Salmonella enterica serovar Typhimurium strains. Our data show extrachromosomal phoQ24 to be dominant over the chromosomal copy of the phoQ gene, conferring the PhoPc phenotype on the recipient strains. In addition, activation of PhoPQ-regulated genes by the plasmid-encoded PhoQ24 did not alter bacterial survival and conferred immunogenicity to the HPV16 VLP expressed in the two S. enterica serovar Typhimurium backgrounds, inducing the production of HPV specific antibodies in mice. This strongly suggests that at least one of the PhoP regulated genes is necessary for mounting an efficient antibody response to HPV16 VLP. This finding sets the stage for further development of a Salmonella-based vaccine against HPV infection and cervical cancer. PMID- 14742518 TI - Sensitized splenocytes result in deleterious cytokine cascade and hyperinflammatory response in rats with Pneumocystis pneumonia despite the presence of corticosteroids. AB - The immune response to the opportunistic pulmonary pathogen Pneumocystis can have beneficial and harmful effects on the host despite the presence of corticosteroids. We hypothesized that this deleterious hyperinflammatory response is associated with exaggerated cytokine production. The adoptive transfer of at least 10(7) immune splenocytes reduced the cyst count in rats with corticosteroid induced pneumocystosis. About 18% of these rats developed clinical illness, an increased lung weight/body weight (LW/BW) ratio, and elevated levels of interleukin 1alpha (IL-1alpha), IL-1beta, IL-6, tumor necrosis factor alpha, IL 5, IL-10, and gamma interferon in the lungs. This hyperinflammatory reaction was not observed in rats that remained clinically well or in control rats. Thus, in this model, corticosteroids have little effect on the cytokine cascade or other adverse effects of the host immune response to Pneumocystis. PMID- 14742519 TI - Acid-responsive gene induction of ammonia-producing enzymes in Helicobacter pylori is mediated via a metal-responsive repressor cascade. AB - Although the adaptive mechanisms allowing the gastric pathogen Helicobacter pylori to survive acid shocks have been well documented, the mechanisms allowing growth at mildly acidic conditions (pH approximately 5.5) are still poorly understood. Here we demonstrate that H. pylori strain 26695 increases the transcription and activity of its urease, amidase, and formamidase enzymes four- to ninefold in response to growth at pH 5.5. Supplementation of growth medium with NiCl2 resulted in a similar induction of urease activity (at low NiCl2 concentration) and amidase activity (at > or = 500 micro M NiCl2) but did not affect formamidase activity. Mutation of the fur gene, which encodes an iron responsive repressor of both amidases, resulted in a constitutively high level of amidase and formamidase activity at either pH but did not affect urease activity at pH 7.0 or pH 5.5. In contrast, mutation of the nikR gene, encoding the nickel responsive activator of urease expression, resulted in a significant reduction of acid-responsive induction of amidase and formamidase activity. Finally, acid responsive repression of fur transcription was absent in the H. pylori nikR mutant, whereas transcription of the nikR gene itself was increased at pH 5.5 in wild-type H. pylori. We hypothesize that H. pylori uses a repressor cascade to respond to low pH, with NikR initiating the response directly via the urease operon and indirectly via the members of the Fur regulon. PMID- 14742520 TI - Investigation of a novel DNase of Streptococcus suis serotype 2. AB - A secreted nuclease, SsnA, was identified in the virulent Streptococcus suis isolate SX332 and subsequently in each of the type strains of capsular serotypes 1 through 9. Screening of 258 porcine clinical isolates from surface (nasal mucosa or palatine tonsil) or internal (joint, brain or other internal organ) locations revealed a significant relationship (P < 0.001) between expression of nuclease and isolation from an internal site. A 3,126-bp gene, ssnA, was identified from a phenotypically nuclease-negative pGh9:ISS1 insertion mutant, and analysis of the predicted SsnA sequence revealed a 35-amino-acid (aa) secretion signal sequence, a 22-aa DNA-binding domain, and a typical gram positive cell wall sorting motif. A requirement of Ca2+ and Mg2+ for SsnA activity was determined, and the substrate specificity was found to be for single and double-stranded linear DNA. Reverse transcription-PCR experiments revealed that ssnA is expressed throughout all stages of S. suis growth, and Western blots with porcine anti-S. suis immune sera against a recombinant, truncated SsnA derivative (rSsnADelta) confirmed that SsnA is expressed in vivo. Furthermore, anti-rSsnADelta antibodies were sufficient to neutralize SsnA activity. Analyses of subcellular fractions of SX332 and derived mutants, on DNA-containing polyacrylamide gels and by Western blotting, suggest that SsnA is cell wall located. PMID- 14742521 TI - Salivary statherin peptide-binding epitopes of commensal and potentially infectious Actinomyces spp. delineated by a hybrid peptide construct. AB - Adhesion of microorganisms to host receptor molecules such as salivary statherin molecules is a common event in oral microbial colonization. Here we used a hybrid peptide construct (with both a hydroxyapatite-binding portion and a test peptide portion) to map the interaction of Actinomyces species (and Candida albicans) with statherin. Adhesion to hybrid peptides and truncated statherin variants revealed three binding types, types I to III. (i) Type I strains of rat, hamster, and human infection origins bound C-terminal-derived QQYTF and PYQPQY peptides. The QQYTF peptide inhibited statherin binding for some strains but not for others. (ii) Type II strains of human and monkey tooth origins bound middle region-derived YQPVPE and QPLYPQ peptides. Neither strain was inhibited by soluble peptides. (iii) Type III strains of human infection origins (and C. albicans) did not bind to either statherin-derived peptides or truncated statherin. Moreover, the type I strains inhibited by QQYTF were also inhibited by TF and QAATF peptides and were detached from statherin by the same peptides. In conclusion, it is suggested that commensal and potentially infectious microorganisms bind middle or C-terminal statherin differently and that other microbes might require discontinuous epitopes. PMID- 14742522 TI - Role of Toll-like receptor 4 in gram-positive and gram-negative pneumonia in mice. AB - To determine the role of Toll-like receptor 4 (TLR4) in the immune response to pneumonia, C3H/HeJ mice (which display a mutant nonfunctional TLR4) and C3H/HeN wild-type mice were intranasally infected with either Streptococcus pneumoniae (a common gram-positive respiratory pathogen) or Klebsiella pneumoniae (a common gram-negative respiratory pathogen). In cases of pneumococcal pneumonia, TLR4 mutant mice showed a reduced survival only after infection with low-level bacterial doses, which was associated with a higher bacterial burden in their lungs 48 h postinfection. In Klebsiella pneumonia, TLR4 mutant mice demonstrated a shortened survival after infection with either a low- or a high-level bacterial dose together with an enhanced bacterial outgrowth in their lungs. These data suggest that TLR4 contributes to a protective immune response in both pneumococcal and Klebsiella pneumonia and that its role is more important in respiratory tract infection caused by the latter (gram-negative) pathogen. PMID- 14742523 TI - Role of the Salmonella pathogenicity island 1 effector proteins SipA, SopB, SopE, and SopE2 in Salmonella enterica subspecies 1 serovar Typhimurium colitis in streptomycin-pretreated mice. AB - Salmonella enterica subspecies 1 serovar Typhimurium (serovar Typhimurium) induces enterocolitis in humans and cattle. The mechanisms of enteric salmonellosis have been studied most extensively in calf infection models. The previous studies established that effector protein translocation into host cells via the Salmonella pathogenicity island 1 (SPI-1) type III secretion system (TTSS) is of central importance in serovar Typhimurium enterocolitis. We recently found that orally streptomycin-pretreated mice provide an alternative model for serovar Typhimurium colitis. In this model the SPI-1 TTSS also plays a key role in the elicitation of intestinal inflammation. However, whether intestinal inflammation in calves and intestinal inflammation in streptomycin-pretreated mice are induced by the same SPI-1 effector proteins is still unclear. Therefore, we analyzed the role of the SPI-1 effector proteins SopB/SigD, SopE, SopE2, and SipA/SspA in elicitation of intestinal inflammation in the murine model. We found that sipA, sopE, and, to a lesser degree, sopE2 contribute to murine colitis, but we could not assign an inflammation phenotype to sopB. These findings are in line with previous studies performed with orally infected calves. Extending these observations, we demonstrated that in addition to SipA, SopE and SopE2 can induce intestinal inflammation independent of each other and in the absence of SopB. In conclusion, our data corroborate the finding that streptomycin-pretreated mice provide a useful model for studying the molecular mechanisms of serovar Typhimurium colitis and are an important starting point for analysis of the molecular events triggered by SopE, SopE2, and SipA in vivo. PMID- 14742524 TI - Immunoglobulin E and eosinophil-dependent protective immunity to larval Onchocerca volvulus in mice immunized with irradiated larvae. AB - Mice immunized with irradiated Onchocerca volvulus third-stage larvae developed protective immunity. Eosinophil levels were elevated in the parasite microenvironment at the time of larval killing, and measurements of total serum antibody levels revealed an increase in the immunoglobulin E (IgE) level in immunized mice. The goal of the present study was to identify the role of granulocytes and antibodies in the protective immune response to the larval stages of O. volvulus in mice immunized with irradiated larvae. Immunity did not develop in mice if granulocytes, including both neutrophils and eosinophils, were eliminated, nor did it develop if only eosinophils were eliminated. Moreover, larvae were killed in naive interleukin-5 transgenic mice, and the killing coincided with an increase in the number of eosinophils and the eosinophil peroxidase (EPO) level in the animals. To determine if EPO was required for protective immunity, mice that were genetically deficient in EPO were immunized, and there were no differences in the rates of parasite recovery in EPO-deficient mice and wild-type mice. Two mouse strains were used to study B-cell function; micro MT mice lacked all mature B cells, and Xid mice had deficiencies in the B-1 cell population. Immunity did not develop in the micro MT mice but did develop in the Xid mice. Finally, protective immunity was abolished in mice treated to eliminate IgE from the blood. We therefore concluded that IgE and eosinophils are required for adaptive protective immunity to larval O. volvulus in mice. PMID- 14742525 TI - Host-pathogen interaction during pneumococcal infection in patients with chronic obstructive pulmonary disease. AB - Acute exacerbation is a frequent complication of chronic obstructive pulmonary disease (COPD). Recent studies suggested a role for bacteria such as Streptococcus pneumoniae in the development of acute exacerbation. For this study, we investigated the following in COPD patients: (i) the epidemiology of pneumococcal colonization and infection, (ii) the effect of pneumococcal colonization on the development of exacerbation, and (iii) the immunological response against S. pneumoniae. We cultured sputa of 269 COPD patients during a stable state and during exacerbation of COPD and characterized 115 pneumococcal isolates by use of serotyping. Moreover, we studied serum immunoglobulin G (IgG) antibody titers, antibody avidities, and functional antibody titers against the seven conjugate vaccine serotypes in these patients. Colonization with only pneumococci (monocultures) increased the risk of exacerbation, with a hazard ratio of 2.93 (95% confidence interval, 1.41 to 6.07). The most prevalent pneumococcal serotypes found were serotypes 19F, 3, 14, 9L/N/V, 23A/B, and 11. We calculated the theoretical coverage for the 7- and 11-valent pneumococcal vaccines to be 60 and 73%, respectively. All patients had detectable IgG levels against the seven conjugate vaccine serotypes. These antibody titers were significantly lower than those in vaccinated healthy adults. Finally, on average, a 2.5-fold rise in serotype-specific and functional antibodies in S. pneumoniae positive sputum cultures was observed during exacerbation. Our data indicate that pneumococcal colonization in COPD patients is frequently caused by vaccine serotype strains. Moreover, pneumococcal colonization is a risk factor for exacerbation of COPD. Finally, our findings demonstrate that COPD patients are able to mount a significant immune response to pneumococcal infection. COPD patients may therefore benefit from pneumococcal vaccination. PMID- 14742526 TI - Myd88-dependent in vivo maturation of splenic dendritic cells induced by Leishmania donovani and other Leishmania species. AB - The usual agent of visceral leishmaniasis in the Old World is Leishmania donovani, which typically produces systemic diseases in humans and mice. L. donovani has developed efficient strategies to infect and persist in macrophages from spleen and liver. Dendritic cells (DC) are sentinels of the immune system. Following recognition of evolutionary conserved microbial products, DC undergo a maturation process and activate antigen-specific naive T cells. In the present report we provide new insights into how DC detect Leishmania in vivo. We demonstrate that in both C57BL/6 and BALB/c mice, systemic injection of L. donovani induced the migration of splenic DC from marginal zones to T-cell areas. During migration, DC upregulated the expression of major histocompatibility complex II and costimulatory receptors (such as CD40, CD80, and CD86). Leishmania induced maturation requires live parasites and is not restricted to L. donovani, as L. braziliensis, L. major, and L. mexicana induced a similar process. Using a green fluorescent protein-expressing parasite, we demonstrate that DC undergoing maturation in vivo display no parasite internalization. We also show that L. donovani-induced DC maturation was partially abolished in MyD88-deficient mice. Taken together, our data suggest that Leishmania-induced DC maturation results from direct recognition of Leishmania by DC, and not from DC infection, and that MyD88-dependent receptors are implicated in this process. PMID- 14742527 TI - Candida albicans yeast and germ tube forms interfere differently with human monocyte differentiation into dendritic cells: a novel dimorphism-dependent mechanism to escape the host's immune response. AB - The ability of Candida albicans to convert from the yeast (Y) form to mycelial forms through germ tube (GT) formation is considered a key feature of the transition of the organism from commensalism to virulence. We show here that human monocytes cultured with granulocyte-macrophage colony-stimulating factor and interleukin-4 (IL-4) after phagocytosis of Y forms did not differentiate into dendritic cells (DCs); they retained CD14, did not acquire CD1a, and were unable to express the maturation markers CD83 and CCR7. Moreover, they did not produce IL-12p70 but secreted IL-10. In addition, they spontaneously expressed high levels of tumor necrosis factor alpha (TNF-alpha), IL-6, and IL-8 mRNA transcripts and were able to induce proliferation of alloreactive memory but not naive T lymphocytes. Conversely, monocytes that had phagocytosed GT forms differentiated into mature CD83+ and CCR7+ DCs; however, there was no up regulation of CD40, CD80, and major histocompatibility complex class II, irrespective of lipopolysaccharide (LPS) treatment. In addition, these cells were unable to produce IL-12 even after LPS stimulation, but they were not functionally exhausted, as shown by their capacity to express TNF-alpha and IL-8 mRNA transcripts. These cells were able to prime naive T cells but not to induce their functional polarization into effector cells. These data indicate that phagocytosis of Y and GT forms has profound and distinct effects on the differentiation pathway of monocytes. Thus, the differentiation of human monocytes into DCs appears to be tunable and exploitable by C. albicans to elude immune surveillance. PMID- 14742528 TI - Identification and cloning of immunodominant antigens of Coxiella burnetii. AB - A sublethal-challenge model was established in BALB/c mice by using protection from the development of severe splenomegaly as an indicator of vaccinogenic activity for evaluation of the protective efficacies of vaccine candidates. To determine the immunodominant antigens as defined by reaction to an infection derived antibody, mouse sera from different stages of experimental infection with various doses of Coxiella burnetii were tested by immunoblotting. Proteins with molecular masses of 14, 16, 21, 28, 32, 45 to 50, 57, and 60 kDa were recognized as immunodominant antigens. Antibody responses in whole-cell antigen (WCA) vaccinated mice were compared with those in unvaccinated mice by immunoblotting using two-dimensional gel-separated C. burnetii antigens. The results indicated that there were significantly different antibody responses during different stages of vaccination and challenge, suggesting that several specific immunogenic antigens may play critical roles in the protection of mice against challenge. To clone these immunogenic antigens, a genomic DNA library of Nine Mile phase I was screened with convalescent-phase antisera from mice. Eighteen novel immunoreactive proteins with molecular masses ranging from approximately 14 to 67 kDa were cloned and identified. Interestingly, several recombinant proteins reacted with sera from both early-stage infected and WCA-vaccinated prechallenged mice. These results suggest that these proteins may play critical roles in the development of protective immunity and that they are logical candidates for vaccine and serodiagnostic reagents. PMID- 14742529 TI - Necrotic death of Rhodococcus equi-infected macrophages is regulated by virulence associated plasmids. AB - Rhodococcus equi is a gram-positive intracellular pathogen that can cause severe bronchopneumonia in foals and AIDS patients. It has been reported that advanced infection of foals is characterized by tissue necrosis, coinciding with the presence of degenerate bacteria-laden macrophages. Here, we report that the possession of the VapA-expressing plasmid, which has been previously correlated with a high level of virulence for foals and mice, strongly increases cytotoxicity of R. equi for murine macrophage-like (J774E) cells. Isolates containing different, VapB-expressing plasmids are less virulent and also have a lower cytotoxic potential. Isogenic strains lacking either plasmid are avirulent and have a very low cytotoxic potential. We show, using fluorescence-activated cell sorter analysis (annexin V/7-amino-actinomycin D and sub-G1-analysis), Western blotting [poly(ADP-ribose) polymerase processing analysis], and electron microscopy (macrophage and nucleus morphologies) that the deaths of murine macrophages are the result of necrotic rather than apoptotic events. We demonstrate that the bacteria must be alive in order to act cytotoxic. Therefore, one effect of the virulence-associated plasmids during infection with R. equi is the promotion of necrotic damage to the host. PMID- 14742530 TI - Characterization of immunoglobulin G and its subclass response to Indian kala azar infection before and after chemotherapy. AB - Serologic parameters of kala-azar were evaluated by Western blot analysis. Sera from kala-azar patients with confirmed diagnoses were screened for immunoglobulin G (IgG) and IgG subclass-specific reactivity against Leishmania donovani membrane antigen (LAg). Heterogeneous LAg-specific IgG reactivity with numerous proteins with molecular masses ranging from 18 to 190 kDa was observed. Though the individual band patterns were varied, seven polypeptides of approximately 31, 34, 51, 63, 72, 91, and 120 kDa were immunoreactive with all the sera tested from kala-azar patients. The band patterns of the immunoblots of sera from patients after treatment and clinical cure with sodium antimony gluconate revealed a decrease in the frequency of the bands. Still, recognition of the 63- and 120-kDa bands was 100%, and the 55- and 91-kDa fractions were recognized in 93% of the sera from cured individuals. Among the IgG subclasses, IgG1 reacted with the greatest number of polypeptides. The 63-kDa protein was again detected by all of the IgG subclasses of all the sera tested. Other fractions recognized by the subclasses of more than 70% of the serum samples included those of 47, 51, 55, and 78 kDa. Following treatment, 63- and 51-kDa bands were the most reactive with the IgG subclasses. LAg-associated cross-reaction with other reference human antisera revealed a mild reactivity of the 63-kDa polypeptide with some of the serum samples from leprosy, malaria, typhoid, tuberculosis, and healthy controls. Western blot analysis of LAg entrapped in liposomes, strong vaccine candidates against experimental visceral leishmaniasis, revealed a more restricted band pattern. The 63-kDa fraction revealed by all pre- and posttreatment sera showed almost negligible levels of cross-reaction with sera from patients with other diseases or from healthy controls. These observations provide insight into induced immunity during kala-azar infection for future application. PMID- 14742531 TI - Caspase-2 and caspase-7 are involved in cytolethal distending toxin-induced apoptosis in Jurkat and MOLT-4 T-cell lines. AB - Cytolethal distending toxin (CDT) from Actinobacillus actinomycetemcomitans is a G2/M cell-cycle-specific growth-inhibitory toxin that leads to target cell distension followed by cell death. To determine the mechanisms by which A. actinomycetemcomitans CDT acts as an immunosuppressive factor, we examined the effects of highly purified CDT holotoxin on human T lymphocytes. Purified CDT was cytolethal toward normal peripheral T lymphocytes that were activated by in vitro stimulation with phytohemagglutinin. In addition, purified CDT showed cytolethal activity against Jurkat and MOLT-4 cells, which are known to be sensitive and resistant, respectively, to Fas-mediated apoptosis. Death in these cell lines was accompanied by the biochemical features of apoptosis, including membrane conformational changes, intranucleosomal DNA cleavage, and an increase in caspase activity in the cells. Pretreatment of Jurkat cells with the general caspase inhibitor z-VAD-fmk mostly suppressed CDT-induced apoptosis. Furthermore, specific inhibitors of caspase-2 and -7 showed significant inhibitory effects on CDT-induced apoptosis in Jurkat cells, and these inhibitory effects were fully associated with reduced activity of caspase-2 or -7 in the CDT-treated Jurkat cells. These results strongly suggest that CDT possesses the ability to induce human T-cell apoptosis through activation of caspase-2 and -7. PMID- 14742532 TI - Evaluation of the association of nine Helicobacter pylori virulence factors with strains involved in low-grade gastric mucosa-associated lymphoid tissue lymphoma. AB - Helicobacter pylori has been associated with the development of two malignant diseases: gastric adenocarcinoma and gastric mucosa-associated lymphoid tissue (MALT) lymphoma. Although the cag pathogenicity island, especially the cagA gene, has been linked with adenocarcinoma, few data concerning H. pylori pathogenic factors involved in low-grade gastric MALT lymphoma are available. The goal of this study was to analyze the prevalence of and correlation between genes coding for seven H. pylori virulence factors (cagA, cagE, vacA, iceA, babA, hopQ, and oipA) and two novel adhesins (sabA and hopZ) by comparing a collection of 43 H. pylori strains isolated from patients with low-grade gastric MALT lymphoma to 39 strains isolated from age-matched patients with gastritis only. Our results show that taken individually, none of the nine genes tested can be considered associated with MALT strains and allow us to conclude that MALT pathogenesis is not linked with more proinflammatory H. pylori strains. We demonstrated that in patients infected with strains harboring the iceA1 allele, sabA functional status, and hopZ "off" status, the odds of developing a MALT lymphoma were 10 times higher. However, the low prevalence of such strains (10 of 43 MALT strains) renders this triple association a low-sensitivity marker for MALT strains. Our data confirmed that H. pylori virulence factors are correlated with one another. If the involvement of H. pylori in MALT lymphoma is well established, the pathomechanism by which gastric lymphoma occurs remains to be identified. PMID- 14742533 TI - Production of secretory immunoglobulin A against Shiga toxin-binding subunits in mice by mucosal immunization. AB - The toxicity of Shiga toxins (Stx) depends on the binding of their B subunits to carbohydrate ligands on host cells. The production of antibodies against B subunits, especially immunoglobulin A (IgA) secreted on the mucosal surface, should contribute to host defense. One of the major problems in attempts to produce IgA against Stx was the poor immunogenicity of B subunits. We were able to produce serum IgA as well as IgG against Stx1B in mice of the H-2d haplotype by means of intranasal immunization with recombinant B subunits of Stx (Stx1B) together with cholera toxin as a mucosal adjuvant. Secretory IgA (S-IgA) was detected in nasal washes but not in feces. We prepared chemically cross-linked Stx1B for use as an immunogen, and the formation of stable oligomers was revealed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and mass spectrometry. When the cross-linked Stx1B was used together with cholera toxin for the intranasal immunization of BALB/c mice, strong enhancement of the immune response was observed. The S-IgA titers in nasal washes were 16- to more than 64 fold higher than those in mice immunized with native Stx1B plus cholera toxin. Furthermore, fecal IgA was detectable when the cross-linked Stx1B was used. The use of cholera toxin was necessary for the induction of high titers of S-IgA in the nasal washes. However, the effect of cross-linking was dependent on the major histocompatibility complex haplotype; that is, no enhancement of IgA production was observed in C57BL/6 mice. The present results provide a practical means of producing IgA against Stx1B in BALB/c mice. PMID- 14742534 TI - RhuR, an extracytoplasmic function sigma factor activator, is essential for heme dependent expression of the outer membrane heme and hemoprotein receptor of Bordetella avium. AB - Genes involved in iron (Fe) acquisition often are regulated in response to the local availability of Fe. In many bacteria, Fe-dependent responsiveness is mediated by Fur, a global Fe-dependent transcriptional repressor. Tighter regulatory control of Fur-responsive genes is afforded by incorporating additional regulators into Fur-dependent regulatory cascades. RhuI, a Fur dependent extracytoplasmic function sigma factor of Bordetella avium, in response to the dual stimulation of Fe starvation and the presence of heme (or hemoproteins), regulates P(bhuR), a heme-responsive promoter which directs expression of the bhuRSTUV heme utilization operon. While BhuR, the outer membrane heme receptor, and RhuI have been shown to be indispensable for heme dependent activation of P(bhuR), collateral components of the regulatory cascade have not been described. In this investigation, RhuR, an integral cytoplasmic membrane protein with homology to anti-sigma factors, is shown to be an essential activator of P(bhuR) expression. The functional domain of RhuR required for heme dependent activation of P(bhuR) expression was mapped to the N-terminal 97 amino acids of the protein by use of a chimeric RhuR-BlaM fusion. Expression of the chimera in a rhuR mutant rendered P(bhuR) constitutive, thereby decoupling the promoter from heme dependency. Growth studies confirmed that B. avium requires RhuR for optimal utilization of hemoglobin, but not hemin, as a sole source of nutrient Fe. These data imply that B. avium expresses, in addition to the BhuR heme/hemoprotein utilization system, an alternative RhuR-independent heme utilization mechanism. A model is proposed in which RhuR is the functional bridge between BhuR and RhuI in a heme-dependent regulatory cascade. PMID- 14742535 TI - Generation of Yersinia pestis attenuated strains by signature-tagged mutagenesis in search of novel vaccine candidates. AB - In a search for novel attenuated vaccine candidates for use against Yersinia pestis, the causative agent of plague, a signature-tagged mutagenesis strategy was used and optimized for a subcutaneously infected mouse model. A library of tagged mutants of the virulent Y. pestis Kimberley53 strain was generated. Screening of 300 mutants through two consecutive cycles resulted in selection of 16 mutant strains that were undetectable in spleens 48 h postinfection. Each of these mutants was evaluated in vivo by assays for competition against the wild type strain and for virulence following inoculation of 100 CFU (equivalent to 100 50% lethal doses [LD50] of the wild type). A wide spectrum of attenuation was obtained, ranging from avirulent mutants exhibiting competition indices of 10(-5) to 10(-7) to virulent mutants exhibiting a delay in the mean time to death or mutants indistinguishable from the wild type in the two assays. Characterization of the phenotypes and genotypes of the selected mutants led to identification of virulence-associated genes coding for factors involved in global bacterial physiology (e.g., purH, purK, dnaE, and greA) or for hypothetical polypeptides, as well as for the virulence regulator gene lcrF. One of the avirulent mutant strains (LD50, >10(7) CFU) was found to be disrupted in the pcm locus, which is presumably involved in the bacterial response to environmental stress. This Kimberley53pcm mutant was superior to the EV76 live vaccine strain because it induced 10- to 100-fold-higher antibody titers to the protective V and F1 antigens and because it conferred efficacious protective immunity. PMID- 14742536 TI - In vivo phase variation and serologic response to lipooligosaccharide of Campylobacter jejuni in experimental human infection. AB - Some Campylobacter jejuni strains which exhibit mimicry of gangliosides in their lipooligosaccharides (LOSs) are associated with development of Guillain-Barre syndrome, which complicates the selection of a suitable C. jejuni strain in a live-attenuated vaccine. C. jejuni 81-176 is the most well characterized strain available, but structurally, LOS of C. jejuni 81-176 exhibits mimicry of predominantly GM2 and GM3 gangliosides. We compared the antiganglioside human serologic responses of 22 volunteers post-oral vaccination (two-dose series, 14 days apart) with a killed whole-cell C. jejuni vaccine, those of volunteers (22 following initial challenge and 5 upon rechallenge) experimentally infected with the homologous C. jejuni vaccine strain 81-176, and those of 12 volunteers used as controls (placebo recipients). All volunteers were evaluated using thin-layer chromatography immuno-overlay and a panel of nine gangliosides at days 0, 21, and 28 either postvaccination or postinoculation. Antiganglioside antibodies were identified at baseline in 6 of the 61 volunteers (9.8%). There were no antiganglioside antibodies observed following vaccination or experimental infection rechallenge. Evidence of seroconversion was observed in 2 of 22 (9.1%) in the initial infection challenge group, comparable to 1 of 12 (8.3%) in the placebo recipients. Additional testing of seven selected volunteers in the initial challenge group at days 0, 3, 7, 10, 21, 28, and 60 showed that when antiganglioside antibodies occurred (mostly anti-GM1 and -GM2), responses were weak and transient. Furthermore, evidence from serologic probing of LOSs of isolates recovered from stools of six volunteers indicated that the isolates had undergone antigenic phase variation in ganglioside mimicry during passage in vivo. Collectively, with the exception of one volunteer with anti-GM2 antibodies at day 60, the results show an absence of persistent antiganglioside antibodies after experimental infection with C. jejuni or following administration of a killed C. jejuni whole-cell oral vaccine, although LOS phase variation occurred. PMID- 14742537 TI - Two studies evaluating the safety and immunogenicity of a live, attenuated Shigella flexneri 2a vaccine (SC602) and excretion of vaccine organisms in North American volunteers. AB - We report the first community-based evaluation of Shigella flexneri 2a strain SC602, a live, oral vaccine strain attenuated by deletion of the icsA (virG) plasmid virulence gene, given at 10(4) CFU. The primary objectives of this trial were to determine the safety and immunogenicity of the vaccine and to determine the duration of colonization. Four of 34 volunteers experienced transient fevers, and three reported diarrhea during the first 3 days of the study. Half of the volunteers mounted a positive serum immunoglobulin A (IgA) response to S. flexneri lipopolysaccharide. All but one of the volunteers excreted the vaccine in their stools for 1 to 33 days, and this excretion was often intermittent. Data from the community-based study were supplemented with an inpatient trial in which three volunteers received 10(3) and nine received 10(4) CFU. All volunteers who received 10(3) CFU excreted SC602 and had an IgA antibody-secreting cell response. Two of these had a serum IgA response. Six of the nine volunteers who received 10(4) CFU excreted SC602. One vaccinee had a transient fever and two met the definition of diarrhea. Six volunteers that received 10(4) CFU had an antibody-secreting cell response, and four had a serum IgA response. SC602 has now been tested at 10(4) CFU in a total of 58 volunteers. The cumulative results of these clinical trials, reported here and previously (Coster et al., Infect. Immun. 67:3437-3443, 1999), have demonstrated that SC602 is a substantially attenuated candidate vaccine that can evoke protection against the most severe symptoms of shigellosis in a stringent human challenge model of disease. PMID- 14742538 TI - Heat shock protein 60 acts as a receptor for the Listeria adhesion protein in Caco-2 cells. AB - The 104-kDa Listeria adhesion protein (LAP) in Listeria monocytogenes is involved in binding to various mammalian cell lines. However, the receptor that interacts with LAP in eukaryotic cells is unknown. In this study, scanning immunoelectron microscopy qualitatively demonstrated greater binding capacity of wild-type (WT) L. monocytogenes strain (F4244) than a LAP-deficient mutant strain (KB208) to Caco-2 cells. The goal of this study was identification of the host cell receptor for LAP. Using a Western blot ligand overlay assay, we identified a protein of 58 kDa to be the putative receptor for LAP from Caco-2 cells. N-terminal sequencing and subsequent database search identified this protein as heat shock protein 60 (Hsp60). Modified immunoseparation with protein A-Sepharose beads bound to the LAP-specific monoclonal antibody H7 (MAb-H7) and a sequential incubation with LAP preparation and Caco-2 lysate confirmed the receptor to be the same 58-kDa protein. Western blot analysis with anti-Hsp60 MAb of whole-cell adhesion between Caco-2 and WT also revealed the receptor protein to be a 58-kDa protein, thus corroborating the identification of Hsp60 as a host cell receptor for LAP. Furthermore, the anti-Hsp60 antibody also caused approximately 74% reduction in binding of L. monocytogenes WT to Caco-2 cells, whereas a control antibody, C11E9, had no effect on binding. The adhesion mechanism of L. monocytogenes to eukaryotic cells is a complex process, and identification of Hsp60 as a receptor for LAP adds to the list of previously discovered ligand-receptor modules that are essential to achieve successful adhesion. PMID- 14742539 TI - Mycobacterium bovis BCG cell wall-specific differentially expressed genes identified by differential display and cDNA subtraction in human macrophages. AB - We have analyzed the gene expression profile of monocytes in response to a highly purified cell wall fraction of Mycobacterium bovis BCG, a clinically approved adjuvant known as BCG cell wall skeleton (BCG-CWS). It is composed of mycolic acid, arabinogalactan, and peptidoglycan and confers Toll-like receptor 2 (TLR2)- and TLR4-dependent signaling that induces monocytes to differentiate into antigen presenting cells (APCs). Here we report differential gene expression analysis with BCG-CWS-stimulated versus nonstimulated monocytes. BCG-CWS exerted massive induction of genes regulated by TLR signaling. Marked gene regulatory characteristics in BCG-CWS-stimulated cells compared to lipopolysaccharide (LPS) stimulated cells follow. (i) Spliced mRNAs encoding soluble forms of TREM-1 and TREM-2 (recently discovered inflammatory-signal-amplifying receptors) were regulated by BCG-CWS, resulting in their differential expression. (ii) The genes for zinc-iron transporter protein (ZIP)-like family proteins HKE-1 and LIV-1 were induced exclusively by BCG-CWS. (iii) Interleukin-23 (IL-23), rather than IL 12p70, was induced by BCG-CWS, while interferon-inducible genes were induced only by LPS. By Northern and reverse transcription-PCR analyses, we confirmed the differential expression of more than 30 BCG-CWS regulatory genes, and their expression was compared with that of LPS and other known TLR ligands. A battery of genes responded rapidly and for a short time to LPS but for a long time to BCG CWS. Structural analysis of the identified novel or hypothetical proteins revealed that some are potential candidates as signaling mediators or transcriptional regulators. Hence, BCG-CWS may profoundly modulate APC responses in a way distinct from that of LPS, leading to possible advantages for its adjuvant-active therapeutic potential. PMID- 14742540 TI - CpG oligodeoxynucleotide and Montanide ISA 51 adjuvant combination enhanced the protective efficacy of a subunit malaria vaccine. AB - Unmethylated CpG dinucleotide motifs present in bacterial genomes or synthetic oligodeoxynucleotides (ODNs) serve as strong immunostimulatory agents in mice, monkeys and humans. We determined the adjuvant effect of murine CpG ODN 1826 on the immunogenicity and protective efficacy of the Saccharomyces cerevisiae expressed 19-kDa C-terminal region of merozoite surface protein 1 (yMSP1(19)) of the murine malaria parasite Plasmodium yoelii. We found that in C57BL/6 mice, following sporozoite challenge, the degree of protective immunity against malaria induced by yMSP1(19) in a formulation of Montanide ISA 51 (ISA) plus CpG ODN 1826 was similar or superior to that conferred by yMSP1(19) emulsified in complete Freund's adjuvant (CFA/incomplete Freund's adjuvant). In total, among mice immunized with yMSP1(19), 22 of 32 (68.7%) with ISA plus CpG 1826, 0 of 4 (0%) with CFA/incomplete Freund's adjuvant, 0 of 4 (0%) with CpG 1826 mixed with ISA (no yMSP1(19)), and 0 of 11 (0%) with CpG 1826 alone were completely protected against development of erythrocytic stage infection after sporozoite challenge. The adjuvant effect of CpG ODN 1826 was manifested as both significantly improved complete protection from malaria (defined as the absence of detectable erythrocytic form parasites) (P = 0.007, chi square) and reduced parasite burden in infected mice. In vivo depletions of interleukin-12 and gamma interferon cytokines and CD4+ and CD8+ T cells in vaccinated mice had no significant effect on immunity. On the other hand, immunoglobulin G (IgG) isotype levels appeared to correlate with protection. Inclusion of CpG ODN 1826 in the yMSP1(19) plus ISA vaccine contributed towards the induction of higher levels of IgG2a and IgG2b (Th1 type) antibodies, suggesting that CpG ODN 1826 caused a shift towards a Th1 type of immune response that could be responsible for the higher degree of protective immunity. Our results indicate that this potent adjuvant formulation should be further evaluated for use in clinical trials of recombinant malarial vaccine candidates. PMID- 14742541 TI - Differential production of systemic and intralesional gamma interferon and interleukin-10 in nodular and ulcerative forms of Buruli disease. AB - Buruli disease, caused by Mycobacterium ulcerans, is the third most important mycobacterial disease in humans besides tuberculosis and leprosy. We have compared systemic and intralesional cytokine production in patients presenting with a nodular form and a necrotizing, ulcerative form of the disease. Gamma interferon (IFN-gamma) levels in response to whole M. ulcerans and Mycobacterium bovis BCG bacilli and in response to purified Ag85 protein from BCG were lower in peripheral blood mononuclear cells (PBMC) cultures from Buruli disease patients than in PBMC from healthy purified protein derivative-positive contacts. Interleukin-4 (IL-4) and IL-13 content was below the detection threshold in these PBMC cultures. IFN-gamma production after stimulation with M. ulcerans was significantly lower (P < 0.05) in PBMC cultures from patients with ulcers than in those from patients with nodules. On the other hand, PBMC from Buruli disease patients produced significant levels of IL-10 in response to M. ulcerans (but not to M. bovis BCG) and production was highest in patients with the ulcerative form. Third, semiquantitative reverse transcription-PCR analysis demonstrated a similar difference in the local, intralesional cytokine profile for the two forms of the disease: high IFN-gamma but low IL-10 mRNA levels in nodular lesions and high IL 10 but low IFN-gamma mRNA levels in ulcerative lesions. Intralesional IL-4 and IL 13 mRNA levels were low and only detected in patients with the ulcerative form. Our results indicate, although they do not formally prove, that production of IL 10 rather than production of IL-4 or IL-13 by Th2-type T cells may be involved in the low M. ulcerans-specific IFN-gamma response in Buruli disease patients. PMID- 14742542 TI - Mechanisms of immunity to Ehrlichia muris: a model of monocytotropic ehrlichiosis. AB - Ehrlichia species can cause life-threatening infections or chronic persistent infections. Mechanisms of protective immunity were examined in an Ehrlichia muris mouse model of monocytotropic ehrlichiosis. C57BL/6 mice possessed strong genetic resistance to E. muris of an undetermined mechanism. CD8 T lymphocytes were particularly important, as revealed by 81% fatalities for E. muris-infected, major histocompatibility complex class I gene knockout mice compared with no deaths for wild-type C3H mice. Moreover, 80% of C3H mice depleted of CD8 and CD4 cells died of E. muris infection compared with only 44% of CD4 cell-depleted mice. CD8 T lymphocytes were demonstrated for the first time in an Ehrlichia infection to exhibit cytotoxic T-lymphocyte activity against Ehrlichia-infected target cells. Both gamma interferon and tumor necrosis factor were shown to play synergistic roles in protective immunity in vivo for the first time, as demonstrated by 75% fatalities when both cytokines were neutralized compared with minimal mortality when they were depleted separately. Passive transfer of antibodies, but not Fab fragments, to E. muris protected C3H/SCID mice against lethal infection. The mechanism of increased susceptibility (22% lethality) of C57BL/6 major histocompatibility complex class II gene knockout mice and CD4 cell depleted C3H mice (i.e., through a gamma interferon or antibody mechanism), as well as the more important role of CD8 T lymphocytes (in the form of cytotoxic T lymphocyte activity and/or gamma interferon production), remains to be elucidated. Protective immunity against monocytotropic E. muris is mediated by a combination of CD8 and CD4 T lymphocytes, gamma interferon, tumor necrosis factor alpha, and antibodies. PMID- 14742543 TI - The staphylococcal ferritins are differentially regulated in response to iron and manganese and via PerR and Fur. AB - Staphylococcus aureus and Staphylococcus epidermidis ferritin (FtnA and SefA, respectively) homologues are antigenic and highly conserved. A previous study showed that ftnA is a component of the S. aureus PerR regulon with its transcription induced by elevated iron and repressed by PerR, which functions as a manganese-dependent transcriptional repressor. We have further investigated the role of iron and Fur in the regulation of PerR regulon genes ftnA (ferritin), ahpC (alkyl-hydroperoxidase), and mrgA (Dps homologue) and shown that iron has a major role in the regulation of the PerR regulon and hence the oxidative stress response, since in the presence of both iron and manganese, transcription of PerR regulon genes is induced above the repressed levels observed with manganese alone. Furthermore the PerR regulon genes are differentially regulated by metal availability and Fur. First, there is an additional level of PerR-independent regulation of ftnA under low-iron conditions which is not observed with ahpC and mrgA. Second, there is a differential response of these genes to Fur as ftnA expression is constitutive in a fur mutant, while ahpC expression is constitutive under low-Fe/Mn conditions but some repression of ahpC still occurs in the presence of manganese, whereas mrgA expression is still repressed in the fur mutant as in wild-type S. aureus, although there is a decrease in the overall level of mrgA transcription. These studies have also shown that FtnA expression is regulated by growth phase, but maximal transcription of ftnA differs dependent on the growth medium. Moreover, there are significant regulatory differences between the S. aureus and S. epidermidis ferritins, as sefA expression in contrast to that of ftnA is derepressed under low-Fe/Mn ion conditions. PMID- 14742544 TI - The Cryptosporidium oocyst wall protein is a member of a multigene family and has a homolog in Toxoplasma. AB - Coccidian parasites are transmitted via a fecal oocyst stage that is exceptionally resistant to environmental stress and harsh chemical treatments, which allows parasites to stably persist outside a host. Because of its oocyst durability Cryptosporidium parvum is a significant water- and food-borne pathogen of humans, as well as animals of agricultural importance. To date, only one apicomplexan oocyst membrane protein has been identified, Cryptosporidium oocyst wall protein 1 (COWP1). COWP1 has a highly cysteine-rich periodicity due to arrays of two apicomplexan-specific motifs, designated the type I and type II domains. In this study, exhaustive BLAST screening of a complete C. parvum genome sequence database resulted in identification of eight additional genes encoding similar arrays of cysteine-rich type I and/or type II domains. Transcript expression analysis revealed that all COWP genes are abundantly expressed at a time when developing oocysts are observed, roughly 48 to 72 h after inoculation of in vitro cultures. A monoclonal antibody recognizing COWP8 specifically localized to the C. parvum oocyst wall, supporting the hypothesis that multiple COWPs play a role in the oocyst wall structure. BLAST screening of the Toxoplasma gondii genome sequence database resulted in identification of a gene encoding at least one COWP homolog (TgOWP1), and this multiexon sequence information was used to isolate a full-length cDNA. Exhaustive screening of Plasmodium sp. genome sequence databases by using COWP genes as BLAST queries failed to detect similar proteins in PLASMODIUM: We therefore propose that the COWP family of proteins have a structural role in apicomplexan species that produce durable shed cysts capable of surviving environmental stress. PMID- 14742545 TI - Enhanced replication of Leishmania amazonensis amastigotes in gamma interferon stimulated murine macrophages: implications for the pathogenesis of cutaneous leishmaniasis. AB - During Leishmania major infection in mice, gamma interferon (IFN-gamma) plays an essential role in controlling parasite growth and disease progression. In studies designed to ascertain the role of IFN-gamma in Leishmania amazonensis infection, we were surprised to find that IFN-gamma could promote L. amazonensis amastigote replication in macrophages (Mphis), although it activated Mphis to kill promastigotes. The replication-promoting effect of IFN-gamma on amastigotes was independent of the source and genetic background of Mphis, was apparently not affected by surface opsonization of amastigotes, was not mediated by interleukin 10 or transforming growth factor beta, and was observed at different temperatures. Consistent with the different fates of promastigotes and amastigotes in IFN-gamma-stimulated Mphis, L. amazonensis-specific Th1 transfer helped recipient mice control L. amazonensis infection established by promastigotes but not L. amazonensis infection established by amastigotes. On the other hand, IFN-gamma could stimulate Mphis to limit amastigote replication when it was coupled with lipopolysaccharides but not when it was coupled with tumor necrosis factor alpha. Thus, IFN-gamma may play a bidirectional role at the level of parasite-Mphi interactions; when it is optimally coupled with other factors, it has a protective effect against infection, and in the absence of such synergy it promotes amastigote growth. These results reveal a quite unexpected aspect of the L. amazonensis parasite and have important implications for understanding the pathogenesis of the disease and for developing vaccines and immunotherapies. PMID- 14742546 TI - Salmonella enterica serovar Typhimurium RamA, intracellular oxidative stress response, and bacterial virulence. AB - Escherichia coli and Salmonella enterica serovar Typhimurium have evolved genetic systems, such as the soxR/S and marA regulons, to detoxify reactive oxygen species, like superoxide, which are formed as by-products of metabolism. Superoxide also serves as a microbicidal effector mechanism of the host's phagocytes. Here, we investigate whether regulatory genes other than soxR/S and marA are active in response to oxidative stress in Salmonella and may function as virulence determinants. We identified a bacterial gene, which was designated ramA (342 bp) and mapped at 13.1 min on the Salmonella chromosome, that, when overexpressed on a plasmid in E. coli or Salmonella, confers a pleiotropic phenotype characterized by increased resistance to the redox-cycling agent menadione and to multiple unrelated antibiotics. The ramA gene is present in Salmonella serovars but is absent in E. coli. The gene product displays 37 to 52% homology to the transcriptional activators soxR/S and marA and 80 to 100% identity to a multidrug resistance gene in Klebsiella pneumoniae and Salmonella enterica serovar Paratyphi A. Although a ramA soxR/S double null mutant is highly susceptible to intracellular superoxide generated by menadione and displays decreased Mn-superoxide dismutase activity, intracellular survival of this mutant within macrophage-like RAW 264.7 cells and in vivo replication in the spleens in Ityr mice are not affected. We concluded that despite its role in the protective response of the bacteria to oxidative stress in vitro, the newly identified ramA gene, together with soxR/S, does not play a role in initial replication of Salmonella in the organs of mice. PMID- 14742547 TI - Mucosal vaccination increases endothelial expression of mucosal addressin cell adhesion molecule 1 in the human gastrointestinal tract. AB - Homing of leukocytes to various tissues is dependent on the interaction between homing receptors on leukocytes and their ligands, addressins, on endothelial cells. Mucosal immunization results in homing of antigen-specific lymphocytes back to the mucosa where they first encountered the antigen. However, it is unknown whether this homing of antigen-specific cells is mediated by an altered endothelial addressin expression after vaccination. Using different immunization routes with an oral cholera vaccine, we show that the endothelial expression of mucosal addressin cell adhesion molecule 1 (MAdCAM-1) is increased in the gastric and upper small intestinal mucosae after immunization through various local routes in the upper gastrointestinal tract. In contrast, rectal immunization did not influence the levels of MAdCAM-1 in the gastric or duodenal mucosa. Furthermore, we show that MAdCAM-1 can be induced on human endothelial cells by tumor necrosis factor alpha (TNF-alpha) and gamma interferon. The vaccine component cholera toxin B subunit (CTB) increased MAdCAM-1 expression on endothelial cells in cultured human gastric explants, an effect that seemed to be mediated by TNF-alpha. In conclusion, MAdCAM-1 expression is increased in the upper gastrointestinal tract after local immunizations with a vaccine containing CTB. This strongly suggests the involvement of MAdCAM-1 in the preferential homing of mucosal lymphocytes to their original site of activation. PMID- 14742548 TI - Members of the merozoite surface protein 7 family with similar expression patterns differ in ability to protect against Plasmodium yoelii malaria. AB - Previously, we described the isolation of the Plasmodium yoelii sequence-related molecules P. yoelii MSP-7 (merozoite surface protein 7) and P. yoelii MSRP-2 (MSP 7-related protein 2) by their ability to interact with the amino-terminal end of P. yoelii MSP-1 in a yeast two-hybrid system. One of these molecules was the homologue of Plasmodium falciparum MSP-7, which was biochemically isolated as part of the shed MSP-1 complex. In the present study, with antibodies directed against recombinant proteins, immunoprecipitation analyses of the rodent system demonstrated that both P. yoelii MSP-7 and P. yoelii MSRP-2 could be isolated from parasite lysates and from parasite culture supernatants. Immunofluorescence studies colocalized P. yoelii MSP-7 and P. yoelii MSRP-2 with the amino-terminal portion of MSP-1 and with each other on the surface of schizonts. Immunization with P. yoelii MSRP-2 but not P. yoelii MSP-7 protected mice against a lethal infection with P. yoelii strain 17XL. These results establish that both P. yoelii MSP-7 and P. yoelii MSRP-2 are expressed on the surface of merozoites and released from the parasite and that P. yoelii MSRP-2 may be the target of a protective immune response. PMID- 14742549 TI - Transcutaneous immunization with combined cholera toxin and CpG adjuvant protects against Chlamydia muridarum genital tract infection. AB - Chlamydia trachomatis is a pathogen of the genital tract and ocular epithelium. Infection is established by the binding of the metabolically inert elementary body (EB) to epithelial cells. These are taken up by endocytosis into a membrane bound vesicle termed an inclusion. The inclusion avoids fusion with host lysosomes, and the EBs differentiate into the metabolically active reticulate body (RB), which replicates by binary fission within the protected environment of the inclusion. During the extracellular EB stage of the C. trachomatis life cycle, antibody present in genital tract or ocular secretions can inhibit infection both in vivo and in tissue culture. The RB, residing within the intracellular inclusion, is not accessible to antibody, and resolution of infection at this stage requires a cell-mediated immune response mediated by gamma interferon-secreting Th1 cells. Thus, an ideal vaccine to protect against C. trachomatis genital tract infection should induce both antibody (immunoglobulin A [IgA] and IgG) responses in mucosal secretions to prevent infection by chlamydial EB and a strong Th1 response to limit ascending infection to the uterus and fallopian tubes. In the present study we show that transcutaneous immunization with major outer membrane protein (MOMP) in combination with both cholera toxin and CpG oligodeoxynucleotides elicits MOMP specific IgG and IgA in vaginal and uterine lavage fluid, MOMP-specific IgG in serum, and gamma interferon-secreting T cells in reproductive tract-draining caudal and lumbar lymph nodes. This immunization protocol resulted in enhanced clearance of C. muridarum (C. trachomatis, mouse pneumonitis strain) following intravaginal challenge of BALB/c mice. PMID- 14742550 TI - Systemic Th1 immunization of mice against Helicobacter pylori infection with CpG oligodeoxynucleotides as adjuvants does not protect from infection but enhances gastritis. AB - Recent reports have suggested that oral vaccination of mice against Helicobacter pylori is dependent on a Th1-mediated immune response. However, oral vaccination in mice neither induces sterilizing immunity nor leads to complete protection from disease. Therefore, in this study we investigated whether a systemic subcutaneous immunization against H. pylori by using CpG oligodeoxynucleotides as a Th1 adjuvant could achieve protection in a mouse model of H. pylori infection. CpG oligodeoxynucleotides are known for their ability to induce nearly entirely Th1-biased immune responses and may be approved for human use in future. Immunization of mice with H. pylori lysate and CpG induced a strong local and systemic Th1 immune response. Despite this strong Th1 response, mice were not protected from infection with H. pylori yet had a 10-fold reduction in the number of H. pylori in the gastric mucosa compared to nonimmunized mice. Of note, reduction of the bacterial density in immunized mice was accompanied by a significantly enhanced gastritis. Hence, systemic Th1 immunization of mice, even though being able to reduce the bacterial load in the stomach, is associated with aggravated pathology. PMID- 14742551 TI - Salmonella enterica serovar Typhimurium requires nonsterol precursors of the cholesterol biosynthetic pathway for intracellular proliferation. AB - We have previously shown that Salmonella enterica serovar Typhimurium infection perturbs the host cholesterol biosynthetic pathway. Here we show that inhibiting the first step of this pathway (3-hydroxy-3-methylglutaryl coenzyme A reductase) reduces the growth of intracellular S. enterica serovar Typhimurium and has no effect on extracellular bacterial growth. Selectively inhibiting synthesis of downstream sterol components has no effect on infection, suggesting that the effect of statins on host nonsterol intermediates is detrimental to bacterial growth. Furthermore, statins also reduce bacterial proliferation in the S. enterica serovar Typhimurium mouse model. This suggests that blocking the production of nonsterol precursors in the host cell can be used to reduce infection. PMID- 14742552 TI - Functional analysis of the cag pathogenicity island in Helicobacter pylori isolates from patients with gastritis, peptic ulcer, and gastric cancer. AB - Helicobacter pylori is the causative agent of a variety of gastric diseases, but the clinical relevance of bacterial virulence factors is still controversial. Virulent strains carrying the cag pathogenicity island (cagPAI) are thought to be key players in disease development. Here, we have compared cagPAI-dependent in vitro responses in H. pylori isolates obtained from 75 patients with gastritis, peptic ulcer, and gastric cancer (n = 25 in each group). AGS gastric epithelial cells were infected with each strain and assayed for (i) CagA expression, (ii) translocation and tyrosine phosphorylation of CagA, (iii) c-Src inactivation, (iv) cortactin dephosphorylation, (v) induction of actin cytoskeletal rearrangements associated with cell elongation, (vi) induction of cellular motility, and (vii) secretion of interleukin-8. Interestingly, we found high but similar prevalences of all of these cagPAI-dependent host cell responses (ranging from 56 to 80%) among the various groups of patients. This study revealed CagA proteins with unique features, CagA subspecies of various sizes, and new functional properties for the phenotypic outcomes. We further showed that induction of AGS cell motility and elongation are two independent processes. Our data corroborate epidemiological studies, which indicate a significant association of cagPAI presence and functionality with histopathological findings in gastritis, peptic ulcer, and gastric cancer patients, thus emphasizing the importance of the cagPAI for the pathogenicity of H. pylori. Nevertheless, we found no significant association of the specific H. pylori-induced responses with any particular patient group. This may indicate that the determination of disease development is highly complex and involves multiple bacterial and/or host factors. PMID- 14742553 TI - Chemokine receptor 5 is dispensable for innate and adaptive immune responses to Listeria monocytogenes infection. AB - Chemokine receptor 5 (CCR5) binds macrophage inflammatory protein 1alpha (MIP 1alpha), MIP-1beta, RANTES, and members of the monocyte chemotactic protein family and is also a receptor for human immunodeficiency virus (HIV). CCR5 ligands can suppress HIV-1 entry into cells. In humans, homozygous mutations of the ccr5 gene confer resistance to HIV-1 infection. The role of CCR5 in defense against microbial infection is unclear. In this study we examined the innate and adaptive immune responses of CCR5-deficient mice to the intracellular bacterial pathogen Listeria monocytogenes. We found that migration of monocytic cells, formation of L. monocytogenes-containing lesions, and bacterial clearance occurred normally in the spleens and livers of CCR5-deficient animals. Activation of macrophages and dendritic cells during the first 3 days postinfection was normal in the absence of CCR5, as demonstrated by intact expression of inducible nitric oxide synthase (iNOS) and production of the cytokines tumor necrosis factor alpha, gamma interferon, and interleukin-12. Priming of L. monocytogenes specific CD8 T cells also occured independently of CCR5 expression. Previously immunized, CCR5-deficient animals mounted normal secondary CD8 T-cell responses and cleared bacteria from infected organs similarly to wild-type controls, suggesting that CCR5 is dispensable for migration and activation of memory CD8 T cells. Our data indicate that CCR5-mediated chemotaxis is not required for defense against infection with L. monocytogenes. PMID- 14742554 TI - Paucibacillary tuberculosis in mice after prior aerosol immunization with Mycobacterium bovis BCG. AB - To develop a murine model of paucibacillary tuberculosis for experimental chemotherapy of latent tuberculosis infection, mice were immunized with viable Mycobacterium bovis BCG by the aerosol or intravenous route and then challenged six weeks later with virulent Mycobacterium tuberculosis. The day after immunization, the counts were 3.71 +/- 0.10 log(10) CFU in the lungs of aerosol immunized mice and 3.65 +/- 0.11 and 4.93 +/- 0.07 log(10) CFU in the lungs and spleens of intravenously immunized mice, respectively. Six weeks later, the lungs of all BCG-immunized mice had many gross lung lesions and splenomegaly; the counts were 5.97 +/- 0.14 and 3.54 +/- 0.07 log(10) CFU in the lungs and spleens of aerosol-immunized mice, respectively, and 4.36 +/- 0.28 and 5.12 +/- 0.23 log(10) CFU in the lungs and spleens of intravenously immunized mice, respectively. Mice were then aerosol challenged with M. tuberculosis by implanting 2.37 +/- 0.13 log(10) CFU in the lungs. Six weeks after challenge, M. tuberculosis had multiplied so that the counts were 6.41 +/- 0.27 and 4.44 +/- 0.14 log(10) CFU in the lungs and spleens of control mice, respectively. Multiplication of M. tuberculosis was greatly limited in BCG-immunized mice. Six weeks after challenge, the counts were 4.76 +/- 0.24 and 3.73 +/- 0.34 log(10) CFU in the lungs of intravenously immunized and aerosol-immunized mice, respectively. In contrast to intravenously immunized mice, there was no detectable dissemination to the spleen in aerosol-immunized mice. Therefore, immunization of mice with BCG by the aerosol route prior to challenge with a low dose of M. tuberculosis resulted in improved containment of infection and a stable paucibacillary infection. This model may prove to be useful for evaluation of new treatments for latent tuberculosis infection in humans. PMID- 14742555 TI - New aspects regarding evolution and virulence of Listeria monocytogenes revealed by comparative genomics and DNA arrays. AB - Listeria monocytogenes is a food-borne bacterial pathogen that causes a wide spectrum of diseases, such as meningitis, septicemia, abortion, and gastroenteritis, in humans and animals. Among the 13 L. monocytogenes serovars described, invasive disease is mostly associated with serovar 4b strains. To investigate the genetic diversity of L. monocytogenes strains with different virulence potentials, we partially sequenced an epidemic serovar 4b strain and compared it with the complete sequence of the nonepidemic L. monocytogenes EGDe serovar 1/2a strain. We identified an unexpected genetic divergence between the two strains, as about 8% of the sequences were serovar 4b specific. These sequences included seven genes coding for surface proteins, two of which belong to the internalin family, and three genes coding for transcriptional regulators, all of which might be important in different steps of the infectious process. Based on the sequence information, we then characterized the gene content of 113 Listeria strains by using a newly designed Listeria array containing the "flexible" part of the sequenced Listeria genomes. Hybridization results showed that all of the previously identified virulence factors of L. monocytogenes were present in the 93 L. monocytogenes strains tested. However, distinct patterns of the presence or absence of other genes were identified among the different L. monocytogenes serovars and Listeria species. These results allow new insights into the evolution of L. monocytogenes, suggesting that early divergence of the ancestral L. monocytogenes serovar 1/2c strains from the serovar 1/2b strains led to two major phylogenetic lineages, one of them including the serogroup 4 strains, which branched off the serovar 1/2b ancestral lineage, leading (mostly by gene loss) to the species Listeria innocua. The identification of 30 L. monocytogenes-specific and several serovar-specific marker genes, such as three L. monocytogenes serovar 4b-specific surface protein-coding genes, should prove powerful for the rapid tracing of listeriosis outbreaks, but it also represents a fundamental basis for the functional study of virulence differences between L. monocytogenes strains. PMID- 14742556 TI - Molecular characterization of a glucose-inhibited division gene, gidA, that regulates cytotoxic enterotoxin of Aeromonas hydrophila. AB - By using a mini-transposon, we obtained two mutated strains of a diarrheal isolate, SSU, of Aeromonas hydrophila that exhibited a 50 to 53% reduction in the hemolytic activity and 83 to 87% less cytotoxic activity associated with the cytotoxic enterotoxin (Act). Act is a potent virulence factor of A. hydrophila and has been shown to contribute significantly to the development of both diarrhea and septicemia in animal models. Subsequent cloning and DNA sequence analysis revealed that transposon insertion occurred at different locations in these two mutants within the same 1,890-bp open reading frame for the glucose inhibited division gene (gidA). A similar reduction in hemolytic (46%) and cytotoxic (81%) activity of Act was noted in the gidA isogenic mutant of A. hydrophila that was generated by marker exchange mutagenesis. Northern blot analysis revealed that the transcription of the cytotoxic enterotoxin gene (act) was not altered in the gidA transposon and isogenic mutants. However, by generating a chromosomal act::alkaline phosphatase gene (phoA) reporter construct, we demonstrated significantly reduced phosphatase activity in these mutants, indicating the effect of glucose-inhibited division (GidA) protein in modulating act gene expression at the translational level. The biological effects of Act in the gidA mutants were restored by complementation. The virulence of the gidA mutants in mice was dramatically reduced compared to the those of the wild type (WT) and complemented strains of A. hydrophila. The histopathological examination of lungs, in particular, indicated severe congestion, alveolar hemorrhage, and acute inflammatory infiltrate in the interstitial compartment and the alveolar spaces when mice were infected with the WT and complemented strains. Minimal-to-mild changes were noted in the lungs with the gidA mutants. Taken together, our data indicate for the first time that GidA regulates the most potent virulence factor of A. hydrophila, Act. PMID- 14742557 TI - Conservation of Babesia bovis small heat shock protein (Hsp20) among strains and definition of T helper cell epitopes recognized by cattle with diverse major histocompatibility complex class II haplotypes. AB - Babesia bovis small heat shock protein (Hsp20) is recognized by CD4+ T lymphocytes from cattle that have recovered from infection and are immune to challenge. This candidate vaccine antigen is related to a protective antigen of Toxoplasma gondii, Hsp30/bag1, and both are members of the alpha-crystallin family of proteins that can serve as molecular chaperones. In the present study, immunofluorescence microscopy determined that Hsp20 is expressed intracellularly in all merozoites. Importantly, Hsp20 is also expressed by tick larval stages, including sporozoites, so that natural tick-transmitted infection could boost a vaccine-induced response. The predicted amino acid sequence of Hsp20 from merozoites is completely conserved among different B. bovis strains. To define the location of CD4+ T-cell epitopes for inclusion in a multiepitope peptide or minigene vaccine construct, truncated recombinant Hsp20 proteins and overlapping peptides were tested for their ability to stimulate T cells from immune cattle. Both amino-terminal (amino acids [aa] 1 to 105) and carboxy-terminal (aa 48 to 177) regions were immunogenic for the majority of cattle in the study, stimulating strong proliferation and IFN-gamma production. T-cell lines from all individuals with distinct DRB3 haplotypes responded to aa 11 to 62 of Hsp20, which contained one or more immunodominant epitopes for each animal. One epitope, DEQTGLPIKS (aa 17 to 26), was identified by T-cell clones. The presence of strain conserved T helper cell epitopes in aa 11 to 62 of the ubiquitously expressed Hsp20 that are presented by major histocompatibility complex class II molecules represented broadly in the Holstein breed supports the inclusion of this region in vaccine constructs to be tested in cattle. PMID- 14742558 TI - Protection against CD95-induced apoptosis by chlamydial infection at a mitochondrial step. AB - Chlamydiae are obligate intracellular bacteria that infect human epithelial and myeloid cells. Previous work has established that chlamydiae are able to protect a cell against apoptosis induced by certain experimentally applied stimuli. Here we provide an analysis of this protective activity against the signal transduction during CD95-induced apoptosis. In HeLa cells overexpressing CD95, infection with Chlamydia trachomatis inhibited the appearance of apoptotic morphology, effector caspase activity, the activation of caspase-9 and -3, and the release of cytochrome c from mitochondria. However, caspase-8-processing and activity (measured as cleavage of Bid) were unaffected by the chlamydial infection. Similarly, infection with the species C. pneumoniae did not prevent the activation of caspase-8 but inhibited the appearance of effector caspase activity upon signaling through CD95. Furthermore, infection with C. trachomatis was able to inhibit CD95-induced apoptosis in Jurkat lymphoid cells, where a mitochondrial contribution is required, but not in SKW6.4 lymphoid cells, where caspase-8 directly activates caspase-3. Taken together, these data show that chlamydial infection can protect cells against CD95-induced apoptosis but only where a mitochondrial signaling step is necessary for apoptotic signal transduction. PMID- 14742559 TI - Gastroenteritis in NF-kappaB-deficient mice is produced with wild-type Camplyobacter jejuni but not with C. jejuni lacking cytolethal distending toxin despite persistent colonization with both strains. AB - Campylobacter jejuni continues to be a leading cause of bacterial enteritis in humans. However, because there are no readily available animal models to study the pathogenesis of C. jejuni-related diseases, the significance of potential virulence factors, such as cytolethal distending toxin (CDT), in vivo are poorly understood. Mice deficient in NF-kappaB subunits (p50(-/-) p65(+/-)) in a C57BL/129 background are particularly susceptible to colitis induced by another enterohepatic microaerobe, Helicobacter hepaticus, which, like C. jejuni, produces CDT. Wild-type C. jejuni 81-176 and an isogenic mutant lacking CDT activity (cdtB mutant) were inoculated into NF-kappaB-deficient (3X) and C57BL/129 mice. Wild-type C. jejuni colonized 29 and 50% of the C57BL/129 mice at 2 and 4 months postinfection (p.i.), respectively, whereas the C. jejuni cdtB mutant colonized 50% of the C57BL/129 mice at 2 p.i. but none of the mice at 4 months p.i. Although the C57BL/129 mice developed mild gastritis and typhlocolitis, they had robust immunoglobulin G (IgG) and Th1-promoted IgG2a humoral responses to both the wild-type strain and the C. jejuni cdtB mutant. In contrast, 75 to 100% of the 3X mice were colonized with both the wild type and the C. jejuni cdtB mutant at similar levels at all times examined. Wild-type C. jejuni caused moderately severe gastritis and proximal duodenitis in 3X mice that were more severe than the gastrointestinal lesions caused by the C. jejuni cdtB mutant. Persistent colonization of NF-kappaB-deficient mice with the wild type and the C. jejuni cdtB mutant was associated with significantly impaired IgG and IgG2a humoral responses (P < 0.001), which is consistent with an innate or adaptive immune system defect(s). These results suggest that the mechanism of clearance of C. jejuni is NF-kappaB dependent and that CDT may have proinflammatory activity in vivo, as well as a potential role in the ability of C. jejuni to escape immune surveillance. NF-kappaB-deficient mice should be a useful model to further study the role of CDT and other aspects of C. jejuni pathogenesis. PMID- 14742560 TI - Antibodies to malaria peptide mimics inhibit Plasmodium falciparum invasion of erythrocytes. AB - Apical membrane antigen 1 (AMA1) is expressed on the surfaces of Plasmodium falciparum merozoites and is thought to play an important role in the invasion of erythrocytes by malaria parasites. To select for peptides that mimic conformational B-cell epitopes on AMA1, we screened a phage display library of >10(8) individual peptides for peptides bound by a monoclonal anti-AMA1 antibody, 4G2dc1, known to inhibit P. falciparum invasion of erythrocytes. The most reactive peptides, J1, J3, and J7, elicited antibody responses in rabbits that recognized the peptide immunogen and both recombinant and parasite AMA1. Human antibodies in plasma samples from individuals exposed to chronic malaria reacted with J1 and J7 peptides and were isolated using immobilized peptide immunoadsorbents. Both rabbit and human antibodies specific for J1 and J7 peptides were able to inhibit the invasion of erythrocytes by P. falciparum merozoites. This is the first example of phage-derived peptides that mimic an important epitope of a blood-stage malaria vaccine candidate, inducing and isolating functional protective antibodies. Our data support the use of J1 and J7 peptide mimics as in vitro correlates of protective immunity in future AMA1 vaccine trials. PMID- 14742561 TI - Platelet-activating factor receptor deficiency delays elimination of adult worms but reduces fecundity in Strongyloides venezuelensis-infected mice. AB - We describe the parasitological kinetics and histopathological and immunological alterations in platelet-activating factor receptor-deficient (PAFR(-/-)) and wild type mice after a single Strongyloides venezuelensis infection (subcutaneous inoculation of 500 L3 larvae). There was no difference in the numbers of worms that reached and became established in the small intestines of PAFR(-/-) and wild type mice. However, at 12 days after infection, significantly more worms were recovered from PAFR(-/-) mice. Although PAFR(-/-) infected mice showed a delay in elimination of adult worms, worms established in the small intestine of these mice produced a significantly lower number of eggs due to a reduction in worm fecundity. There were also significant reductions in the number of circulating and tissue eosinophils and tumor necrosis factor levels in the small intestines of PAFR(-/-) mice infected for 7 days compared to the number and level in wild type mice. Histological analysis confirmed the reduced inflammatory process and revealed that the PAFR(-/-) mice had a smaller number of goblet cells. The concentrations of the type 2 cytokines interleukin-4 (IL-4), IL-5, and IL-10 were lower in small intestine homogenates and in supernatants of antigen-stimulated lymphocytes from spleens or mesenteric lymph nodes of PAFR(-/-) mice than in the corresponding preparations from wild-type mice. Thus, in S. venezuelensis infected PAFR(-/-) mice, decreased intestinal inflammation is associated with enhanced worm survival but decreased fecundity. We suggest that although a Th2 predominant inflammatory response decreases worm survival, the worm may use factors produced during this response to facilitate egg output and reproduction. PAFR-mediated responses appear to modulate these host-derived signals that are important for worm fecundity. PMID- 14742562 TI - Haemophilus ducreyi strain ATCC 27722 contains a genetic element with homology to the vibrio RS1 element that can replicate as a plasmid and confer NAD independence on haemophilus influenzae. AB - The nucleotide sequence of pNAD1, a plasmid from Haemophilus ducreyi identified on the basis of its ability to confer NAD independence on Actinobacillus pleuropneumoniae and H. influenzae, has been determined. In addition to containing the nadV gene, the plasmid contains homologues of the rstR and rstA genes, genes encoding repressor and replication proteins, respectively, in the Vibrio CTXphi and the Vibrio RS1 element, suggesting a single-stranded bacteriophage origin for pNAD1. Tandem copies of the plasmid are integrated into the H. ducreyi 35000HP genome. PMID- 14742563 TI - Rapid accumulation of eosinophils in lung lesions in guinea pigs infected with Mycobacterium tuberculosis. AB - Guinea pig eosinophils were positively identified in bronchoalveolar lavage populations and in the lung granulomas of Mycobacterium tuberculosis-infected guinea pigs. It is possible that the rapid influx of these cells, and their subsequent degranulation during acute pulmonary tuberculosis, may play a key role in the susceptibility of this animal model. PMID- 14742564 TI - Type III secretion: a virulence factor delivery system essential for the pathogenicity of Burkholderia mallei. AB - By creating mutations in the Burkholderia mallei ATCC 23344 animal pathogen-like type III secretion system (TTSS), this study analyzes the correlation between type III secretion and the pathogenicity of ATCC 23344 in vivo. Mutagenesis demonstrated that a functional TTSS was required for the full pathogenicity of ATCC 23344 in the BALB/c mouse and Syrian hamster models of infection. However, vaccination with each mutant failed to elicit a protective immunity against challenge with wild-type ATCC 23344. PMID- 14742565 TI - The ferritin-like Dps protein is required for Salmonella enterica serovar Typhimurium oxidative stress resistance and virulence. AB - Resistance to phagocyte-derived reactive oxygen species is essential for Salmonella enterica serovar Typhimurium pathogenesis. Salmonella can enhance its resistance to oxidants through the induction of specific genetic pathways controlled by SoxRS, OxyR, sigma(S), sigma(E), SlyA, and RecA. These regulons can be found in a wide variety of pathogenic and environmental bacteria, suggesting that evolutionarily conserved mechanisms defend against oxidative stress both endogenously generated by aerobic respiration and exogenously produced by host phagocytic cells. Dps, a ferritin-like protein found in many eubacterial and archaebacterial species, appears to protect cells from oxidative stress by sequestering iron and limiting Fenton-catalyzed oxyradical formation. In Escherichia coli and some other bacterial species, Dps has been shown to accumulate during stationary phase in a sigma(S)-dependent fashion, bind nonspecifically to DNA, and form a crystalline structure that compacts and protects chromatin from oxidative damage. In the present study, we provide evidence that Dps protects Salmonella from iron-dependent killing by hydrogen peroxide, promotes Salmonella survival in murine macrophages, and enhances Salmonella virulence. Reduced numbers of dps mutant bacteria in the livers and spleens of infected mice are consistent with a role of Dps in protecting Salmonella from oxidative stress encountered during infection. PMID- 14742566 TI - Heat shock protein 60 is the major antigen which stimulates delayed-type hypersensitivity reaction in the macaque model of Chlamydia trachomatis salpingitis. AB - Chlamydial delayed-type hypersensitivity antigens were analyzed by using the subcutaneous salpingeal autotransplant model of Macaca nemestrina infected with Chlamydia trachomatis serovar E. Heat shock protein 60 was the only antigen shown to induce delayed-type hypersensitivity among other antigens tested, including UV inactivated organisms, recombinant major outer membrane protein, purified outer membrane proteins, and heat shock protein 10. PMID- 14742567 TI - Evidence for acquisition of the lipooligosaccharide biosynthesis locus in Campylobacter jejuni GB11, a strain isolated from a patient with Guillain-Barre syndrome, by horizontal exchange. AB - Campylobacter jejuni GB11, a strain isolated from a patient with Guillain-Barre syndrome, has been shown to be genetically closely related to the completely sequenced strain C. jejuni NCTC 11168 by various molecular typing and serotyping methods. However, we observed that the lipooligosaccharide (LOS) biosynthesis genes strongly diverged between GB11 and NCTC 11168. We sequenced the LOS biosynthesis locus of GB11 and found that it was nearly identical to the class A LOS locus from the C. jejuni HS:19 Penner serotype strain (ATCC 43446). Analysis of the DNA sequencing data showed that a horizontal exchange event involving at least 14.26 kb had occurred in the LOS biosynthesis locus of GB11 between galE (Cj1131c in NCTC 11168) and gmhA (Cj1149 in NCTC 11168). Mass spectrometry of the GB11 LOS showed that GB11 expressed an LOS outer core that mimicked the carbohydrate portion of the gangliosides GM1a and GD1a, similar to C. jejuni ATCC 43446. The serum from the GB11-infected patient was shown to react with the LOS from both GB11 and ATCC 43446 but not with that from NCTC 11168. These data indicate that the antiganglioside response in the GB11-infected patient was raised against the structures synthesized by the acquired class A LOS locus. PMID- 14742568 TI - Periodontitis vaccine decreases local prostaglandin E2 levels in a primate model. AB - Interleukin-1beta, tumor necrosis factor alpha, prostaglandin E2 (PGE2), and Porphyromonas gingivalis-specific immunoglobulin G levels in gingival crevicular fluid were measured in primates immunized with a P. gingivalis vaccine followed by ligature-induced periodontitis. Only PGE2 levels were dramatically suppressed (P < 0.0001) in immunized animals versus controls. A significant correlation (P < 0.027) was also found between PGE2 levels and decreased bone loss scores. This study presents the first evidence of a potential mechanism involved in periodontitis vaccine-induced suppression of bone loss in a nonhuman primate model and offers insight into the role of PGE2 in periodontal destruction. PMID- 14742569 TI - Activated THP-1 cells: an attractive model for the assessment of intracellular growth rates of Mycobacterium tuberculosis isolates. AB - Capacity of certain Mycobacterium tuberculosis isolates to grow more rapidly in human macrophages may be indicative of increased virulence. Significant differences were observed in intracellular growth of two isolates from sites of tuberculosis transmission, with an outbreak-associated strain growing faster than a strain causing disease in only one person. Activated THP-1 cells are a suitable alternative to peripheral blood monocyte models. PMID- 14742570 TI - An msbB homologue carried in plasmid pO157 encodes an acyltransferase involved in lipid A biosynthesis in Escherichia coli O157:H7. AB - Escherichia coli O157:H7 carries a chromosomal msbB1 and a plasmid-encoded msbB2 gene. We characterized msbB2 function as a homologue of msbB1 by examination of wild-type organisms and mutant strains that lacked functional msbB1, msbB2, and both msbB1 and msbB2. The msbB double-mutant strain generated pentaacyl lipid A, while the single-mutant strains synthesized hexaacyl lipid A. Complementation with overexpressed msbB2 converted pentaacyl into hexaacyl lipid A in the double mutant strain. The transcription of both msbB genes occurred simultaneously. Lack of MsbB2 activity slightly increased the microheterogeneity of the lipid A species. These results suggest that the msbB2 gene plays a role not only in the routine generation of fully hexaacylated lipid A but also in suppressing the microheterogeneity of lipid A species, the endotoxic determinant of the organism. PMID- 14742571 TI - Needle-free skin patch vaccination method for anthrax. AB - Three immunizations of mice with recombinant protective antigen (rPA) by transcutaneous immunization (TCI) induced long-term neutralizing antibody titers that were superior to those obtained with aluminum-adsorbed rPA. In addition, rPA alone exhibited adjuvant activity for TCI. Forty-six weeks after completion of TCI, 100% protection was observed against lethal anthrax challenge. PMID- 14742572 TI - Intranasal immunization of mice with group B streptococcal protein rib and cholera toxin B subunit confers protection against lethal infection. AB - Intranasal immunization of mice with Rib, a cell surface protein of group B streptococcus (GBS), conjugated to or simply coadministered with the recombinant cholera toxin B subunit, induces systemic immunoglobulin G (IgG) and local IgA antibody responses and confers protection against lethal GBS infection. These findings have implications for the development of a human GBS vaccine. PMID- 14742573 TI - Downregulation of the DNA-binding activity of nuclear factor-kappaB p65 subunit in Porphyromonas gingivalis fimbria-induced tolerance. AB - Porphyromonas gingivalis fimbriae induce high levels of nuclear factor-kappaB (NF kappaB)-dependent cytokine release upon primary but not secondary stimulation of monocytic cells (FimA tolerance). In this study, fimbriae induced Toll-like receptor-mediated activation of both p50 and p65 subunits of NF-kappaB upon primary cellular activation. However, activation of the transactivating p65 subunit (but not of the transcriptionally inactive p50 subunit) was significantly inhibited in fimbria-restimulated cells. Moreover, expression of a NF-kappaB dependent reporter gene was inhibited upon secondary stimulation with fimbriae. NF-kappaB p65 downregulation may thus contribute to induction of FimA tolerance. PMID- 14742574 TI - Hydrogen peroxide-mediated killing of Caenorhabditis elegans: a common feature of different streptococcal species. AB - Recently, we reported that Streptococcus pyogenes kills Caenorhabditis elegans by the use of hydrogen peroxide (H2O2). Here we show that diverse streptococcal species cause death of C. elegans larvae in proportion to the level of H2O2 produced. H2O2 may mask the effects of other pathogenicity factors of catalase negative bacteria in the C. elegans infection model. PMID- 14742575 TI - Identification of five outer membrane-associated proteins among cross-protective factor proteins of Pasteurella multocida. AB - Fowl cholera is caused by Pasteurella multocida serovars A:1, A:3, and A:4. The 39-kDa cross-protective factor protein and four other membrane proteins of the membrane proteome of P. multocida were identified. We determined that the 39-kDa cross-protective protein was Pasteurella lipoprotein B, or PlpB. PMID- 14742576 TI - Comparison of Campylobacter jejuni isolates implicated in Guillain-Barre syndrome and strains that cause enteritis by a DNA microarray. AB - We asked whether Campylobacter jejuni isolated from patients with Guillain-Barre syndrome (GBS) differ from isolates isolated from patients with uncomplicated gastrointestinal infection using DNA microarray analysis. We found that specific GBS genes or regions were not identified, and microarray analysis confirmed significant genomic heterogeneity among the isolates. PMID- 14742577 TI - Haemophilus influenzae porin induces Toll-like receptor 2-mediated cytokine production in human monocytes and mouse macrophages. AB - The production of proinflammatory cytokines is likely to play a major pathophysiological role in meningitis and other infections caused by Haemophilus influenzae type b (Hib). Previous studies have shown that Hib porin contributes to signaling of the inflammatory cascade. We examined here the role of Toll-like receptors (TLRs) and the TLR-associated adaptor protein MyD88 in Hib porin induced production of tumor necrosis factor alpha (TNF-alpha) and interleukin-6 (IL-6). Hib porin-induced TNF-alpha and IL-6 production was virtually eliminated in macrophages from TLR2- or MyD88-deficient mice. In contrast, macrophages from lipopolysaccharide (LPS)-hyporesponsive C3H/HeJ mice, which are defective in TLR4 function, responded normally to Hib porin. Moreover anti-TLR2 antibodies but not anti-TLR4 antibodies significantly reduced Hib porin-stimulated TNF-alpha and IL 6 release from the human monocytic cell line THP-1. These data indicate that the TLR2/MyD88 pathway plays an essential role in Hib porin-mediated cytokine production. These findings may be useful in the development of alternative therapies aimed at reducing excessive inflammatory responses during Hib infections. PMID- 14742578 TI - The bacterial insertion sequence element IS256 occurs preferentially in nosocomial Staphylococcus epidermidis isolates: association with biofilm formation and resistance to aminoglycosides. AB - Staphylococcus epidermidis is a normal constituent of the healthy human microflora, but it is also the most common cause of nosocomial infections associated with the use of indwelling medical devices. Isolates from device associated infections are known for their pronounced phenotypic and genetic variability, and in this study we searched for factors that might contribute to this flexibility. We show that mutator phenotypes, which exhibit elevated spontaneous mutation rates, are rare among both pathogenic and commensal S. epidermidis strains. However, the study revealed that, in contrast to those of commensal strains, the genomes of clinical S. epidermidis strains carry multiple copies of the insertion sequence IS256, while other typical staphylococcal insertion sequences, such as IS257 and IS1272, are distributed equally among saprophytic and clinical isolates. Moreover, detection of IS256 was found to be associated with biofilm formation and the presence of the icaADBC operon as well as with gentamicin and oxacillin resistance in the clinical strains. The data suggest that IS256 is a characteristic element in the genome of multiresistant nosocomial S. epidermidis isolates that might be involved in the flexibility and adaptation of the genome in clinical isolates. PMID- 14742579 TI - The siderophore receptor IroN, but not the high-pathogenicity island or the hemin receptor ChuA, contributes to the bacteremic step of Escherichia coli neonatal meningitis. AB - Using a neonatal rat meningitis model, we examined the involvement of three iron uptake systems, namely, the high-pathogenicity island, the hemin receptor ChuA, and the siderophore receptor IroN, in the pathogenesis of Escherichia coli neonatal meningitis. Only IroN appeared to play a major role during the bacteremic step of the disease. PMID- 14742580 TI - Cortical Lewy body disease. PMID- 14742581 TI - Hemicraniectomy after massive hemispheric cerebral infarction: are we ready for a prospective randomised controlled trial? PMID- 14742582 TI - Visualisation of the circumventricular organs by fluorescence endoscopy. PMID- 14742583 TI - How to spot bias and other potential problems in randomised controlled trials. PMID- 14742584 TI - The importance of suspecting superficial siderosis of the central nervous system in clinical practice. AB - Once the central nervous system surface is greatly encrusted with haemosiderin, even removing the source of bleeding will have little effect on the progression of clinical deterioration. Superficial siderosis of the central nervous system is rare and insidious, but magnetic resonance imaging has turned a previously late, mainly autoptical diagnosis into an easy, specific, in vivo, and possibly early one. Avoiding long diagnostic delay will be very important in those cases susceptible of causal treatment. PMID- 14742585 TI - Premorbid proneness to distress and episodic memory impairment in Alzheimer's disease. AB - BACKGROUND: Chronic stress has been associated with impaired episodic memory, but the association of premorbidly experienced distress with memory function in Alzheimer's disease is unknown. OBJECTIVE: To investigate the link between proneness to distress and Alzheimer's disease. METHODS: Participants were 363 persons with clinically diagnosed Alzheimer's disease. At baseline, a knowledgeable informant rated each person's premorbid personality (that is, before dementia onset) along five dimensions, one of which was the tendency to experience psychological distress. Participants underwent structured clinical evaluations at baseline and then annually for up to four years. Each evaluation included 17 cognitive tests from which previously established measures of episodic memory, visuoconstruction, repetition, and naming were derived. RESULTS: In a series of random effects models adjusted for age, sex, and education, premorbid distress proneness was associated with baseline impairment in episodic memory but not with impairment in other cognitive domains, or with change in any cognitive domain. No other trait was related to baseline function or rate of decline in any cognitive domain. CONCLUSIONS: The results suggest that premorbid proneness to experience psychological distress is related to level of impairment in episodic memory in persons with Alzheimer's disease, but neither distress proneness nor other personality traits are related to disease progression. PMID- 14742586 TI - Walking difficulties in patients with Alzheimer's disease might originate from gait apraxia. AB - OBJECTIVES: To investigate whether gait apraxia is a possible cause for some of the walking abnormalities shown by patients with Alzheimer's disease. METHODS: 60 patients with Alzheimer's disease, selected as being free from overt extrapyramidal impairment or other potential causes of walking deficits, were assessed with a new test evaluating aspects of walking and related movements. Norms for this test were collected from a sample of 182 healthy volunteers. RESULTS: 40% of the Alzheimer group performed below the cut off score on this test, and half performed poorly. Performance of the Alzheimer group in the walking skills test correlated highly with scores in a test assessing limb apraxia and with dementia severity. CONCLUSIONS: Gait apraxia may be the cause of walking disorders found in a subgroup of patients with Alzheimer's disease. Its detection is made easier by the use of a standardised test, but still relies heavily on the exclusion of other causes of walking deficits. It is a recognisable and independent form of apraxia. PMID- 14742588 TI - Subthalamic nucleus stimulation induces deficits in decoding emotional facial expressions in Parkinson's disease. AB - BACKGROUND: Bilateral subthalamic nucleus (STN) stimulation is recognised as a treatment for parkinsonian patients with severe levodopa related motor complications. Although adverse effects are infrequent, some behavioural disturbances have been reported. OBJECTIVE: To investigate the consequences of STN stimulation on emotional information processing in Parkinson's disease by assessing the performance of an emotional facial expression (EFE) decoding task in a group of patients before and after surgery. METHODS: 12 non-demented patients with Parkinson's disease were studied. They were assessed one month before surgery and three months after. Their ability to decode EFEs was assessed using a standardised quantitative task. Overall cognitive function, executive function, visuospatial perception, depression, and anxiety were also measured. Twelve healthy controls were matched for age, sex, and duration of education. RESULTS: Before surgery, the patients showed no impairment in EFE decoding compared with the controls. Their overall cognitive status was preserved but they had a moderate dysexecutive syndrome. Three months after surgery, they had significant impairment of EFE decoding. This was not related to their overall cognitive status or to depression/anxiety scores. Visuospatial perception was not impaired. There was no change in the extent of the dysexecutive syndrome except for a reduction in phonemic word fluency. CONCLUSIONS: Bilateral STN stimulation disturbs negative emotional information processing in Parkinson's disease. The impairment appears specific and unrelated to certain secondary variables. This behavioural complication of STN may have implications for the patient's social life. PMID- 14742590 TI - Rate and correlates of weight change in Huntington's disease. AB - OBJECTIVE: To determine the rate and correlates of weight change in a large, well characterised sample of patients with Huntington's disease followed at 44 sites by the Huntington Study Group. PARTICIPANTS AND METHODS: Weight change was assessed in 927 adults with a definite diagnosis of Huntington's disease who were followed prospectively for (mean (SD)) 3.4 (1.4) years. The unified Huntington's disease rating scale was used to assess weight, motor dysfunction (including chorea and dystonia), depressive symptoms, and functional decline. RESULTS: Random effects modelling determined that patients gained an average of 0.11 (1.7) kg/year and their chorea scores increased by 0.36 (0.78) points/year. There were significant but weak relations between weight loss and increasingly severe chorea (r = -0.13), worse baseline motor performance (r = -0.12), less severe baseline depressed mood (r = 0.14), and poorer baseline independence ratings (r = 0.07). Patients who were within 0 to 2 years of symptom onset at the time of the baseline visit gained more weight than those with longer disease duration. CONCLUSIONS: Weight loss following symptom onset is not a consistent feature of Huntington's disease. The mechanisms contributing to weight change in this condition are unclear and probably multifactorial. Future studies examining asymptomatic carriers of the mutation could be helpful in identifying incipience of low body weight and may be better suited for identifying clinical correlates of weight loss than studies in symptomatic patients. PMID- 14742591 TI - Topography of cerebral atrophy in early Huntington's disease: a voxel based morphometric MRI study. AB - OBJECTIVES: To analyse grey matter changes in early stages of Huntington's disease using magnetic resonance imaging (MRI) and the technique of voxel based morphometry (VBM). METHODS: Forty four patients with a molecularly confirmed clinical diagnosis of Huntington's disease based on the presence of motor signs were included in the study. Patients were clinically rated using the Unified Huntington's Disease Rating Scale; all were in early clinical stages of the disease (that is, Shoulson stages I and II). High resolution volume rendering MRI scans (MP-RAGE) were acquired. MRI data were volumetrically analysed in comparison to an age matched normal database by VBM, using statistical parametric mapping (SPM99). RESULTS: In Huntington's disease, robust regional decreases in grey matter density (p<0.001, corrected for multiple comparisons)-that is, atrophy-were found bilaterally in striatal areas as well as in the hypothalamus and the opercular cortex, and unilaterally in the right paracentral lobule. The topography of striatal changes corresponded to the dorso-ventral gradient of neuronal loss described in neuropathological studies. Stratification according to clinical severity showed a more widespread involvement extending into the ventral aspects of the striatum in the group of more severely affected patients. CONCLUSIONS: The topography of cerebral volume changes associated with Huntington's disease can be mapped using VBM. It can be shown that cerebral grey matter changes co-vary with clinical severity and CAG repeat length. PMID- 14742592 TI - Corpus callosum signal intensity in patients with bipolar and unipolar disorder. AB - BACKGROUND: Anatomical abnormalities in the corpus callosum have been reported in magnetic resonance imaging (MRI) studies in patients with bipolar but not unipolar disorder. MRI signal intensity can be used as a putative index of corpus callosum myelination. OBJECTIVES: To measure MRI signal intensity in patients with bipolar and unipolar disorder to investigate abnormalities of corpus callosum myelination. METHODS: The study involved 29 DSM-IV bipolar patients (mean (SD) age, 35 (11) years; 16 male, 13 female), 23 DSM-IV unipolar patients (41 (10) years; 4 male, 19 female), and 36 healthy controls (37 (10) years; 23 male, 13 female). A 1.5T GE Signa magnet was employed, with a fast spin echo sequence. Corpus callosum signal intensity was obtained blindly using the semiautomated software NIH Image 1.62. RESULTS: Bipolar patients had lower corpus callosum signal intensity for all callosal subregions (genu, anterior and posterior body, isthmus, splenium) than healthy controls (ANCOVA, age and sex as covariates, p<0.05). No significant differences were found between unipolar and healthy subjects (ANCOVA, age and sex as covariates, p>0.05). CONCLUSIONS: The findings suggest abnormalities in corpus callosum white matter in bipolar but not unipolar patients, possibly because of altered myelination. Such abnormalities could lead to impaired interhemispheric communication in bipolar disorder. Longitudinal MRI studies involving first episode and early onset bipolar patients will be necessary for a better understanding of the potential role of abnormalities of corpus callosum myelination in the pathophysiology of bipolar disorder. PMID- 14742594 TI - Correlation of anxiety and depression symptoms in patients with restless legs syndrome: a population based survey. AB - BACKGROUND AND OBJECTIVES: Restless legs syndrome (RLS) is an important and common cause of insomnia, and previous studies indicate that psychiatric wellbeing may be impaired among RLS patients. We aimed to investigate the interaction between anxiety/depression and RLS in a population based survey. METHODS: Data were drawn from the Mersin University Neuro-Epidemiology Project, a representative community sample of adults aged over 17 years residing in Mersin (n = 3234). Subjects found to be positive for RLS (n = 103) were evaluated for symptoms of anxiety and depression using the Hamilton Anxiety and Depression Scales and compared with the same number of contemporaneous control subjects. RESULTS: Significantly greater anxiety and depression symptoms were observed among patients with RLS than in the control subjects. Our data also seem to provide initial evidence of a correlation between the severity of RLS and of anxiety and depression symptoms (r = 0.21, p = 0.03 and r = 0.201, p = 0.04 respectively). CONCLUSIONS: Assessment of psychiatric status of RLS patients can be helpful and sometimes necessary to determine additional features and treatment strategies of this bothering condition. Further studies are needed to replicate our findings using longitudinal data. PMID- 14742595 TI - Clinical study of 35 patients with dysarthria-clumsy hand syndrome. AB - OBJECTIVES: Although dysarthria-clumsy hand syndrome (DCHS) is a well known and infrequent lacunar syndrome, there are few data regarding the spectrum of associated clinical characteristics, anatomical site of lesion, and aetiopathogenetic mechanisms. We report a clinical description of this subtype of lacunar stroke based on data collected from a prospective acute stroke registry. METHODS: From 2500 acute stroke patients included in a hospital based prospective stroke registry over a 12-year period, 35 patients were identified as having DCHS. RESULTS: DCHS accounted for 1.6% of all acute stroke patients (35/2110), 1.9% of acute ischaemic stroke (35/1840), and 6.1% of lacunar syndromes (35/570) admitted consecutively to a neurology department and included in the stroke registry over this period. The results supported the lacunar hypothesis in 94.3% of patients (n = 33). Atherothrombotic and cardioembolic infarction occurred in only one patient each (2.9%). No patient with DCHS had an intracerebral haemorrhage. Outcome was good (mortality in hospital 0%, symptom free at discharge 45.7%). After multivariate analysis, absence of limitation at discharge, limb weakness but not cerebellar-type ataxia, and internal capsule (40%), pons (17%), and corona radiata (8.6%) location were significantly associated with DCHS. CONCLUSIONS: DCHS is a rare cerebrovascular syndrome, and supports the criteria of the lacunar hypothesis. The majority of patients in this study had internal capsule infarcts. The prognosis is good with striking similarity compared with other types of lacunar strokes. There are important differences between DCHS and non-lacunar strokes. Internal capsule and pons are the most frequent cerebral sites. PMID- 14742596 TI - Visuospatial abilities in cerebellar disorders. AB - BACKGROUND: Cerebellar involvement in spatial data management has been suggested on experimental and clinical grounds. OBJECTIVE: To attempt a specific analysis of visuospatial abilities in a group of subjects with focal or atrophic cerebellar damage. METHODS: Visuospatial performance was tested using the spatial subtests of the WAIS, the Benton line orientation test, and two tests of mental rotation of objects-the Minnesota paper form board test (MIN) and the differential aptitude test (DAT). RESULTS: In the Benton line orientation test, a test of sensory analysis and elementary perception, no deficits were present in subjects with cerebellar damage. In MIN, which analyses the capacity to process bidimensional complex figures mentally, and in the DAT, which is based on mental folding and manipulation of tridimensional stimuli, subjects with cerebellar damage were impaired. CONCLUSIONS: The results indicate that lesions of the cerebellar circuits affect visuospatial ability. The ability to rotate objects mentally is a possible functional substrate of the observed deficits. A comparison between visuospatial performance of subjects with focal right and left cerebellar lesions shows side differences in the characteristics of the visuospatial syndrome. Thus cerebellar influences on spatial cognition appear to act on multiple cognitive modules. PMID- 14742597 TI - Interrater agreement of the diagnosis and classification of a first seizure in childhood. The Dutch Study of Epilepsy in Childhood. AB - OBJECTIVE: To assess the interrater agreement of the diagnosis and the classification of a first paroxysmal event in childhood. METHODS: The descriptions of 100 first paroxysmal events were submitted to two panels each consisting of three experienced paediatric neurologists. Each observer independently made a diagnosis based on clinical judgment and thereafter a diagnosis based on predefined descriptive criteria. Then, the observers discussed all patients within their panel. The agreement between the six individual observers was assessed before discussion within each panel and after that, between the two panels. RESULTS: Using their clinical judgement, the individual observers reached only fair to moderate agreement on the diagnosis of a first seizure (mean (SE) kappa 0.41 (0.03)). With use of defined descriptive criteria the mean (SE) kappa was 0.45 (0.03). The kappa for agreement between both panels after intra-panel discussion increased to 0.60 (0.06). The mean (SE) kappa for the seizure classification by individual observers was 0.46 (0.02) for clinical judgment and 0.57 (0.03) with use of criteria. After discussion within each panel the kappa between the panels was 0.69 (0.06). In 24 out of 51 children considered to have had a seizure, agreement was reached between the panels on a syndrome diagnosis. However, the epileptic syndromes were in most cases only broadly defined. CONCLUSIONS: The interrater agreement on the diagnosis of a first seizure in childhood is just moderate. This phenomenon hampers the interpretation of studies on first seizures in which the diagnosis is only made by one observer. The use of a panel increased the interrater agreement considerably. This approach is recommended at least for research purposes. Classification into clinically relevant syndromes is possible only in a very small minority of children with a single seizure. PMID- 14742598 TI - Voxel based morphometry reveals a distinct pattern of frontal atrophy in progressive supranuclear palsy. AB - BACKGROUND: Frontal lobe atrophy is a well known neuropathological feature of progressive supranuclear palsy (PSP), accompanied by characteristic neuropsychological deficits. OBJECTIVE: To determine subregional frontal lobe atrophy patterns in patients with PSP using voxel based morphometry (VBM). METHODS: VBM is an observer unbiased volumetry which allows the investigation of the entire brain. An optimised protocol for normalisation, segmentation, and correction for volume changes in preprocessing was used. Grey matter, white matter, and cerebrospinal fluid (CSF) partitions in 12 patients with probable PSP were compared with 12 healthy controls matched for age and sex. RESULTS: In PSP patients, the following cortical areas were decreased in volume (p(corr)<0.05): the prefrontal cortex, predominantly the medial frontal gyri and a cluster in the left lateral middle frontal gyrus; the insular region including the frontal opercula; both supplementary motor areas; and the left medio-temporal area (V5). White matter comparisons revealed a volume reduction in both frontotemporal regions and the mesencephalon. Analysis of the CSF compartment showed no significant regional changes between the groups. CONCLUSIONS: Frontal atrophy in PSP predominantly involves mesio-frontal targets of striatal projections. This atrophy pattern probably accounts for cardinal PSP associated behavioural deficits. PMID- 14742599 TI - Topodiagnostic investigations on the sympathoexcitatory brain stem pathway using a new method of three dimensional brain stem mapping. AB - OBJECTIVES: To study the incompletely understood sympathoexcitatory pathway through the human brain stem, using a new method of three dimensional brain stem mapping on the basis of digitally postprocessed magnetic resonance imaging (MRI). METHODS: 258 consecutive patients presenting with acute signs of brain stem ischaemia underwent biplane T2 and EPI diffusion weighted MRI, with slice orientation parallel and perpendicular to a transversal slice selection of the stereotactic anatomical atlas of Schaltenbrand and Wahren, 1977. The individual slices were digitally normalised and projected onto the appropriate slices of the anatomical atlas. For correlation analysis lesions were imported into a three dimensional model of the human brain stem. RESULTS: 31 of the 258 patients had Horner's syndrome caused by acute brain stem ischaemia. Only four of the patients with Horner's syndrome had pontine infarctions, 12 had pontomedullary lesions, and 15 had medullary lesions. Correlation analysis showed significantly affected voxels in the dorsolateral medulla but not in the pons. A statistical comparison with infarct topology in patients with medullary lesions but without Horner's syndrome indicated that involvement of the medial and ventral part of affected voxels located in the ventrolateral medullary tegmentum was specific for Horner's syndrome. CONCLUSIONS: Based on this first in vivo topodiagnostic study, the central sympathoexcitatory pathway probably descends through the dorsal pons before converging on specific generators in the ventrolateral medullary tegmentum at a level below the IX and X nerve exits. PMID- 14742600 TI - The use of evoked potentials for clinical correlation and surgical outcome in cervical spondylotic myelopathy with intramedullary high signal intensity on MRI. AB - OBJECTIVE: To investigate the use of motor evoked potentials (MEPs) and somatosensory evoked potentials (SEPs) for clinical significance and surgical outcome in patients with cervical spondylotic myelopathy (CSM) with intramedullary high signal intensity on T2 weighted MRI. METHODS: Forty nine patients were scored according to the modified Japanese Orthopaedic Association (JOA) score for cervical myelopathy. MEP and SEP studies were performed and the results were categorised as normal or abnormal. Thirty nine patients who had received surgical decompression were re-evaluated after 6 months. Surgical outcome was represented by the recovery ratio of the JOA score. RESULTS: Abnormal MEPs were observed in 44 patients (arm: 43; leg: 30). Abnormal SEPs were found in 32 patients: (median: 24; tibial: 23). Patients with abnormal SEPs had a worse JOA score than those with normal SEPs. Thirty nine patients received surgical treatment. Patients younger than 55 had better recovery ratios than those who were 55 or older (p = 0.005, two sample Student's t test). Patients with normal median SEPs also had better recovery ratios than those with abnormal median SEPs (p = 0.007, two sample Student's t test). Among median SEP variables, only N9-20 was significantly associated with recovery ratio (p = 0.016, stepwise linear regression), with age factor controlled (p = 0.025, stepwise linear regression). CONCLUSION: Arm MEP was the most sensitive EP test for detecting myelopathy in patients with chronic CSM. Median and tibial SEPs correlated well with the severity of myelopathy while normal median SEPs correlated with good surgical outcome. Among median SEP variables, only N9-20 correlated with surgical outcome. PMID- 14742601 TI - A novel mutation of myelin protein zero associated with an axonal form of Charcot Marie-Tooth disease. AB - OBJECTIVE: To report a new mutation in the MPZ gene which encodes myelin protein zero (P0), associated with an axonal form of Charcot-Marie-Tooth disease (CMT). METHODS: Three patients from an Italian family with a mild, late onset axonal peripheral neuropathy are described clinically and electrophysiologically. To detect point mutation in MPZ gene the whole coding sequence was examined. The structure of the mutated protein was investigated using the three dimensional model of P0. RESULTS: All patients showed a relatively mild CMT phenotype characterised by late onset and heterogeneity of the clinical and electrophysiological features. Molecular analysis demonstrated a novel heterozygous T/A transversion in the exon 3 of MPZ gene that predicts an Asp109Glu amino acid substitution in the extracellular domain of the P0. Asp109 is found at the protein surface, on beta strand E, in the interior of the P0 tetramer. CONCLUSIONS: The identification of Asp109Glu mutation confirms the pivotal role of P0 in axonal neuropathies and stresses the phenotypic heterogeneity associated with MPZ mutations. This study suggests the value of screening for MPZ mutations in CMT family members with minor clinical and electrophysiological signs of peripheral neuropathy. PMID- 14742602 TI - The multiple sclerosis impact scale (MSIS-29) is a reliable and sensitive measure. AB - OBJECTIVE: The aim of this study was to assess the psychometric properties of the Multiple Sclerosis Impact Scale (MSIS-29) for patients in the community and in a hospital setting. METHODS: During an epidemiological study, 172 people with multiple sclerosis (MS) were examined and completed the MSIS-29, the London Handicap Scale, and Beck's Depression Inventory; disability was assessed by the Kurtzke Expanded Disability Status Score (EDSS) and the Multiple Sclerosis Functional Composite. At the hospital neurology clinic, 102 MS patients completed the MSIS-29 and EDSS assessments were performed; 41 of these patients had repeat evaluations six months later. The psychometric properties of the MSIS-29 were examined. RESULTS: In the 172 community and the 102 hospital patients the psychometric properties of the MSIS-29 were satisfactory, with high convergent and low divergent validity. It was significantly responsive to change in the contexts of self-reported change (p<0.034) and EDSS worsening (p<0.001). The MSIS 29 physical score did not change over time when the EDSS was stable, and increased significantly in proportion to EDSS deterioration (p = 0.014). CONCLUSIONS: The psychometric properties of the MSIS-29 are acceptable; it is a valuable outcome measure in intervention studies of patients with MS. PMID- 14742603 TI - Outcome and prognostic factors of hemicraniectomy for space occupying cerebral infarction. AB - OBJECTIVE: To determine long term functional outcome and length of survival of patients undergoing decompressive craniectomy for space occupying infarction of the middle cerebral artery (MCA), and to identify risk factors associated with death and unfavourable outcomes METHODS: Databases of patients undergoing decompressive craniectomy for space occupying MCA infarction compiled at eight neurosurgical departments (1996-2001) were merged, and 188 patients were evaluated. Mortality was calculated by the Kaplan-Meier method. Clinical outcome was rated using the Glasgow outcome scale (GOS). The prognostic impact of patient related covariates on length of survival and the GOS was analysed multivariately. RESULTS: The unadjusted 3, 6, and 12 month mortality rates were 7.9%, 37.6%, and 43.8%, respectively (median follow up, 26 weeks). In the "best" multivariate model, age >50 years (p<0.02) and the involvement of two or more additional vascular territories (p<0.01) had an unfavourable impact on length of survival. The adjusted six month mortality was as low as 20.0% (no risk factor) and as high as 59.7% (two risk factors). A GOS score of 50 years (p<0.0003): 34.9% of the patients 3, as compared with 12.0% of the elderly subpopulation. The side of the infarct did not have prognostic relevance. CONCLUSIONS: Results of surgical treatment in patients <50 years of age undergoing decompressive craniectomy are encouraging. The effectiveness of decompressive craniectomy for patients >50 years remains questionable and should be analysed in the framework of a prospective randomised study. PMID- 14742604 TI - Sudomotor function in familial dysautonomia. AB - BACKGROUND: Patients with familial dysautonomia (FD) manifest episodic hyperhidrosis despite the reduction of sudomotor fibres and sweat glands associated with this autonomic neuropathy. We assessed peripheral sudomotor nerve fibre and sweat gland function to determine if this symptom was due to peripheral denervation hypersensitivity. METHODS: In 14 FD patients and 11 healthy controls, direct and axon reflex mediated sweat responses were determined by measuring transepidermal water loss (TEWL) after application of acetylcholine via a microdialysis membrane, a novel method to evaluate sudomotor function in neuropathy patients. Results were compared with data from conventional quantitative sudomotor axon reflex testing (QSART). Using microdialysis, interstitial fluid was analysed for plasma proteins to evaluate protein extravasation induced by acetylcholine as an additional parameter of C-fibre function. RESULTS: Although reduced axon reflex sweating was expected in FD patients, neither direct or axon reflex mediated sweat responses, nor acetylcholine induced protein extravasation differed between control and patient groups. However, the baseline resting sweat rate was higher in FD patients than controls (p<0.05). TEWL and QSART test results correlated (r = 0.64, p = 0.01), proving the reliability of TEWL methodology in evaluating sudomotor function. CONCLUSION: The finding of normal direct and axon reflex mediated sweat output in FD patients supports our hypothesis that, in a disorder with severe sympathetic nerve fibre reduction, sudomotor fibres, but not the sweat gland itself, exhibit chemical hypersensitivity. This might explain excessive episodic hyperhidrosis in situations with increased central sympathetic outflow. PMID- 14742605 TI - False negative findings in intraoperative SEP monitoring: analysis of 658 consecutive neurosurgical cases and review of published reports. AB - OBJECTIVES: To determine the sensitivity of intraoperative monitoring in neurosurgical operations using somatosensory evoked potentials and to identify reasons for false negative findings and possible settings with an increased risk for monitoring failure. METHODS: SEP monitoring of 658 neurosurgical operations was analysed. The target of monitoring was the function of a hemisphere in 251 cases, the brain stem in 198 cases, and the spinal cord in 209 cases. RESULTS: In 27 cases (4.1%), monitoring was classified as false negative. Further analysis showed that five of these patients had experienced delayed neurological damage. Among the remaining 22 false negative cases, 14 had a minor neurological deficit and eight had severe neurological damage. Overall sensitivity and negative predictive value of SEP monitoring was 79% and 96%, respectively. For the detection of severe neurological damage the corresponding figures were 91% and 98%. Sensitivity of monitoring varied depending on the target of monitoring and the type of lesion. Monitoring was less likely to detect neurological damage in surgery for infratentorial tumours with brain stem compression, small lesions of the motor cortex, and small vessel damage during aneurysm surgery. CONCLUSIONS: SEP monitoring has acceptable sensitivity for detecting neurological damage during different neurosurgical procedures. Distinct settings with an increased risk of monitoring failure can be identified. In these cases measures to enhance the sensitivity of monitoring should be considered. PMID- 14742606 TI - Micturition disturbance in acute idiopathic autonomic neuropathy. AB - OBJECTIVE: To define the nature of micturition disturbance in patients with acute idiopathic autonomic neuropathy (AIAN). METHODS: Micturitional symptoms were observed during hospital admissions and the in outpatient clinics in six patients with clinically definite AIAN (typical form in four, cholinergic variant in one, autonomic-sensory variant in one). Urodynamic studies included medium-fill water cystometry, external sphincter electromyography, and a bethanechol test. RESULTS: Four patients had urinary retention and two had voiding difficulty as the initial presentation. Patients with retention became able to urinate within a week (two to seven days). The major symptoms at the time of urodynamic studies (three weeks to four months after disease onset in most cases) were voiding difficulty and nocturnal frequency. None had urinary incontinence. Complete recovery from the micturition disturbance took from three months to >18 years. The recovery period was shorter in a patient with cholinergic variant, and it was longer in two patients who had a longer duration of initial urinary retention. Micturition disturbance tended to improve earlier than orthostatic hypotension. The major urodynamic abnormalities were detrusor areflexia on voiding (5), denervation supersensitivity to bethanechol (3); low compliance detrusor (1); and impaired bladder sensation (2). None had neurogenic motor unit potentials of the external sphincter muscles. CONCLUSIONS: In patients with AIAN, urinary retention and voiding difficulty are common initial presentations. The underlying mechanisms seem to be pre- and postganglionic cholinergic dysfunction with preservation of somatic sphincter function. The bladder problems tend to improve earlier than orthostatic hypotension. PMID- 14742608 TI - Prediction of treatment response to rivastigmine in Alzheimer's dementia. AB - OBJECTIVES: To predict the treatment response to rivastigmine in patients with Alzheimer's dementia using neuropsychological and EEG data. METHODS: A neuropsychological examination and a quantitative EEG study were done in 20 patients with Alzheimer's dementia before initiating treatment with rivastigmine. After one week of treatment a second EEG examination was done. Therapeutic efficacy was determined six months after treatment initiation. Treatment response was defined as improvement in short term memory after six months of rivastigmine treatment. RESULTS: For the group of patients as a whole, there was a significant improvement in short term memory and orientation during rivastigmine treatment. The mini-mental state score improved from 20.2 to 21.7 (NS). In the EEG, theta power decreased significantly after one week of treatment. Treatment responders had a greater decrease in theta power after one week of treatment and a better short term memory at baseline than non-responders. Decrease in theta power during rivastigmine treatment and baseline short term memory were good predictors of treatment response. CONCLUSIONS: Generally available neuropsychological and EEG data may be useful for predicting response to rivastigmine in patients with Alzheimer's disease. PMID- 14742609 TI - Low dose quetiapine for drug induced dyskinesias in Parkinson's disease: a double blind cross over study. AB - OBJECTIVE: Drug induced dyskinesias remain a challenging problem in the long term management of Parkinson's disease (PD). We have assessed the effect of quetiapine on dyskinesias in a double blind placebo controlled cross over study. METHODS: Nine patients with PD were enrolled and received 25 mg of quetiapine or placebo at night for two weeks in prerandomised order, with one week of wash out between treatment periods. Assessments were made using on-off diaries, self assessment of dyskinesias, and L-dopa challenges at baseline and after each treatment period. Videotapes were rated blindly by two raters using modified Abnormal Involuntary Movement Scale and Goetz scores. Patients subsequently went on open label quetiapine at 50 mg/day, for a mean duration of 30 days, and completed the same self assessment forms. RESULTS: During the double blind phase, no significant change in dyskinesias was found on either 25 mg of quetiapine or placebo. Duration of off states and Unified PD Rating Scale motor scores also remained unchanged. Moderate tiredness and daytime sleepiness occurred in two patients on quetiapine. One patient dropped out early for unrelated reasons. Eight patients completed the open label phase. On 50 mg/day of quetiapine, a slight reduction in dyskinesias occurred on some scales. Reduction in dyskinesia severity on visual analogue scales was by 50.1%. Off time was not significantly increased. This improvement was not strongly reflected in patients' overall impression of treatment effect. Drowsiness and daytime sleep episodes led to discontinuation in four patients, after completion of the study, and two additional patients stopped treatment after the study because of lack of effect. CONCLUSION: Our study failed to demonstrate an antidyskinetic effect of low dose (25 mg) quetiapine. The absence of an increase in parkinsonism combined with a possible antidyskinetic effect on higher doses warrants further investigation. PMID- 14742612 TI - Tangier disease--a diagnostic challenge in countries endemic for leprosy. AB - A case of Tangier disease (TD) is reported from India. The patient had presented with indolent mononeuritis multiplex and trophic ulcers of 16 years duration mimicking Hansen's disease. He received antileprosy treatment for one and a half years. Nerve conduction studies revealed features of demyelinating neuropathy. Biopsies of the sural nerve and skin showed striking vacuolation of Schwann cells and myelin sheaths, and foamy vacuolated fibroblasts, respectively, and no evidence of Hansen's disease. Low levels of apolipoprotein A1 (ApoA1) and cholesterol in the serum and undetectable levels of high density lipoprotein (HDL) and low density lipoprotein (LDL) cholesterol in the blood confirmed the diagnosis of TD. This is the first reported case of TD from a tropical country India. An attempt to establish a correct diagnosis should be made by demonstrating the histopathological and lipoprotein abnormality to avoid long term medications that are chosen empirically and are unnecessary. The importance of recognising this disease in a country where Hansen's disease is highly endemic cannot be overemphasised. PMID- 14742610 TI - Vascular endothelial growth factor (VEGF) is increased in serum, but not in cerebrospinal fluid in HIV associated CNS diseases. AB - Vascular endothelial growth factor (VEGF) is a potent angiogenic and mitogenic peptide, which also induces several mediators that may play a role in HIV induced CNS damage. VEGF levels were determined in cerebrospinal fluid (CSF) and serum samples from patients with (n = 8) and without (n = 19) directly HIV associated CNS disorders and HIV negative control patients (n = 18). VEGF serum but not CSF levels were significantly increased in HIV infected patients with (381.1 (78.9) pg/ml) HIV associated CNS diseases compared with those without (120.8 (13.1) pg/ml) and HIV negative control patients (133.1(14.8) pg/ml). Serum samples from patients with untreated HIV associated encephalopathy (HIVE, n = 3) contained the highest VEGF levels (583.9 (71.5) pg/ml). In two patients VEGF serum levels were reduced during antiretroviral therapy. However, regardless of effective viral suppression, patients with HIVE still had higher levels compared with HIV infected patients without HIVE. A relevant increase of serum VEGF was not observed in patients without HIVE though high HI viral load. We conclude that HIVE is associated with increased serum VEGF levels. Further studies are warranted to elucidate the role of VEGF in HIVE. PMID- 14742613 TI - Multiple microembolic borderzone brain infarctions and endomyocardial fibrosis in idiopathic hypereosinophilic syndrome and in Schistosoma mansoni infestation. AB - We report two cases of multiple distal borderzone ischaemic strokes associated with hypereosinophilia due to idiopathic hypereosinophilic syndrome in one patient and to acute Schistosoma mansoni infestation in the other. Endomyocardial fibrosis (EMF) was documented pathologically, in one case at autopsy and in the other after cardiac surgery; and by cardiac CT, with initially negative echocardiography, in one patient. These observations suggest that so called borderzone infarcts may be due to microembolisms and that, in the context of hypereosinophilia, EMF diagnosis warrants complete cardiac investigation including cardiac CT and repeat echocardiography. PMID- 14742614 TI - Detection of oligoclonal free kappa chains in the absence of oligoclonal IgG in the CSF of patients with suspected multiple sclerosis. AB - BACKGROUND: Oligoclonal free kappa bands are present as frequently as oligoclonal IgG bands in the cerebrospinal fluid (CSF) from patients with definite multiple sclerosis (MS) and can even occur in the absence of oligoclonal IgG. As such, they too are markers of an ongoing intrathecal immune process. OBJECTIVES: To determine how frequently oligoclonal free kappa bands are detectable in the CSF from patients with clinical signs and symptoms suggestive of MS in the absence of CSF restricted oligoclonal IgG. METHODS: An immunoaffinity mediated immunoblotting technique specific for free kappa chains was used, after isoelectric focusing of paired CSF and serum samples from 33 patients with clinical signs and symptoms suggestive of MS but without CSF oligoclonal IgG. CSF data were correlated with MRI results in the context of the new diagnostic criteria from McDonald et al. RESULTS: Eighteen CSF samples contained oligoclonal free kappa bands (54%), mainly from patients with motor dysfunction (83%) and optic neuritis (64%). All patients with a positive MRI according to Barkhof's criteria (n = 6) had free kappa bands in their CSF. CONCLUSIONS: (1) Oligoclonal free kappa bands in the CSF are related to the dissemination of MS lesions; (2) such bands should be looked for in oligoclonal IgG negative CSF, and (3) the presence of free kappa bands in the CSF may be a substitute for oligoclonal IgG in the McDonald's criteria for diagnosis of MS. PMID- 14742615 TI - Isolated pulmonary arteriovenous fistula without Rendu-Osler-Weber disease as a cause of cryptogenic stroke. AB - There has been uncertainty as to whether a right to left shunt through an isolated pulmonary arteriovenous fistula (P-AVF) without Rendu-Osler-Weber (ROW) disease can cause paradoxical brain embolism. A population of 747 acute ischaemic stroke patients was examined to determine the frequency and clinical characteristics of those patients who had an isolated P-AVF. The presence of a P AVF was determined as follows. On patients with a stroke of undetermined cause, both transoesophageal echocardiography and transcranial Doppler with saline contrast medium was performed to detect a right to left shunt. If a P-AVF was then suspected, selective pulmonary angiography and enhanced chest CT was performed to confirm the presence of the P-AVF. Four patients (0.5%) were diagnosed as having a stroke associated with an isolated P-AVF. All the patients were middle-aged women (mean age 61 years). In all these patients, the P-AVF could not have been suspected on physical findings or chest x ray. The P-AVF was always single and located in the lower lobe. All the patients had asymptomatic deep venous thrombosis, and three patients developed pulmonary embolism. As D dimer and thrombin-antithrombin complex were elevated in all patients, this indicated an activation of both fibrinolytic and thrombin activity. Our results show that an isolated P-AVF without ROW disease can cause paradoxical brain embolism. Thus, the existence of an isolated P-AVF as a right to left shunt in patients with a stroke of unknown origin should not be overlooked, even if a P AVF is not suggested by the initial physical findings or chest x ray. PMID- 14742616 TI - The use of contact lenses to treat visually symptomatic congenital nystagmus. AB - It has been suggested that contact lens wear improves the visual function of patients with visual loss from congenital nystagmus. In this study, four patients with congenital nystagmus had two evaluations separated by at least one week (one with spectacles, one with contact lenses) including visual acuity, contrast sensitivity, oscillopsia scale, quality of life questionnaire (NEI VFQ-25), and eye movement recording with an infrared tracking system. All patients subjectively preferred contact lenses to spectacles. Their contrast sensitivity and VFQ-25 scores were improved with contact lenses compared with spectacles alone. Several parameters of nystagmus showed no change in two patients, worsening in one patient and improvement in one patient. This suggests that much of the clinical improvement observed in our patients may result from a better optical correction of their refractive error with contact lenses than with spectacles, rather than from a true damping effect of the nystagmus by contact lenses. PMID- 14742617 TI - Activity patterns of leg muscles in periodic limb movement disorder. AB - The movements of leg muscles in reference to periodic limb movement disorder (PLMD) have only been described in global terms. The sequences of contracting muscles that cause the PLMs are said to be stereotypical. There is, however, doubt about this fixed sequencing in PLMD. Our goal was to define the sequence of muscle movements in PLMs and then analyse their patterns. We recorded with surface EMG all movements of the muscles said to be involved in PLMs (extensor digitorum brevis, EDB; tibialis anterior, TA; biceps femoris, BF; tensor fasciae latae; TFL) as well as the quadriceps (Q) and soleus (S) muscles in 12 patients with restless legs syndrome combined with PLMD. Accompanying polysomnography provided the sleep parameters. In total, 469 movements were analysed. In only 12% was there the appearance of the classic movement (EDB-TA-BF-TFL) or its direct variants. The most frequent sequences were characterised by contraction of only the TA, TA-EDB only, or TA-EDB followed by all other combinations (32%). The pattern EDB only, EDB-TA, or EDB-TA followed by contraction of one or more other muscles, was seen in 18%. All other combinations appeared in much smaller numbers or only once. Eight patients had specific patterns. Three consistently started with the same muscle. One patient always contracted all six muscles. Six patients never contracted more than three muscles. The number of muscles contracted correlated positively with the appearance of arousal from sleep. The interval between onset of contractions within the PLMs varied randomly in a range of 0-1 s. Within PLMs many variations of muscle movements were documented. Patterns were recognisable, individually determined, and related to arousal from sleep. PMID- 14742618 TI - Barre-Lieou "syndrome". PMID- 14742619 TI - No benefit derived from repetitive transcranial magnetic stimulation in depression: a prospective, single centre, randomised, double blind, sham controlled "add on" trial. AB - Repetitive transcranial magnetic stimulation (rTMS) has been reported to demonstrate slight effects in the treatment of depression. Hence, a novel bilateral versus unilateral and sham stimulation design was applied to further assess rTMS' antidepressant effects. Forty one medication free patients with major depression, admitted to a psychiatric unit specialising in affective disorders, were consecutively randomised into 3 groups. Group A1 (n = 12) received unilateral active stimulation consisting of high frequency (hf) rTMS over the left dorsolateral prefrontal cortex (LDLPC) and subsequent sham low frequency (lf) rTMS over the right dorsolateral prefrontal cortex (RDLPC). Group A2 (n = 13) received simultaneous bilateral active stimulation consisting of hf rTMS over the LDLPC and lf-rTMS over the RDLPC. Group C (n = 13) received bilateral sham stimulation. Stimulation was performed on 10 consecutive workdays. All patients received antidepressant medication on the first day of stimulation, which was continued during and after the stimulation period. As no significant difference in antidepressant outcome between group A1 and A2 was found, the two groups were pooled. The time course of the outcome variables Hamilton depression rating scale (HDRS(21)) and Beck depression inventory (days 0, 7, 14, 28) by repeated measures analysis of variance revealed no significant group differences (in terms of a group by time interaction), whereas there was a significant effect of time on all three outcome variables in all groups. The results suggest that rTMS as an "add on" strategy, applied in a unilateral and a bilateral stimulation paradigm, does not exert an additional antidepressant effect. PMID- 14742620 TI - Collateral brain damage, a potential source of cognitive impairment after selective surgery for control of mesial temporal lobe epilepsy. AB - BACKGROUND: Highly selective epilepsy surgery in temporal lobe epilepsy is intended to achieve seizure freedom at a lower cognitive risk than standard en bloc resections, but bears the risk of collateral cortical damage resulting from the surgical approach. OBJECTIVE: To investigate cortical damage associated with selective amygdalo-hippocampectomy (SAH). METHODS: 34 epileptic patients were evaluated. They were randomly assigned to SAH using either a sylvian (9 left/10 right) or a transcortical surgical approach (5 left/10 right). Postoperative MRI signal intensity changes adjacent to the approach were correlated with performance changes in serial word and design list learning. RESULTS: Losses in verbal learning and recognition memory were positively related to signal intensity changes, independent of the side of the resection, the surgical approach, or the extent of the mesial resection. Losses in consolidation/retrieval (memory) were greater after left sided surgery. Losses in design learning were related to right sided surgery and signal intensity changes. Seizure outcome (85% seizure-free) did not differ depending on the side or type of surgery. CONCLUSIONS: Collateral damage to cortical tissues adjacent to the surgical approach contributes to postoperative verbal and figural memory outcome after SAH. Controlling for collateral damage may clarify the controversial memory outcomes after SAH reported by different surgical centres. PMID- 14742621 TI - Oral contraceptive induced chorea: another condition associated with anti-basal ganglia antibodies. AB - Use of oral contraceptives is a recognised but infrequent cause of chorea. This type of chorea has usually been considered a reactivation of Sydenham's chorea by an unknown mechanism. A patient developed a chorea triggered by the use of oral contraceptives with no definite evidence of previous Sydenham's chorea or recent streptoccocal infections. However, the patient had positive anti-basal ganglia antibodies, which supports an immunological basis for the pathophysiology of this chorea. PMID- 14742622 TI - Characteristics of dystonic movements in primary and symptomatic dystonias. AB - OBJECTIVE: To compare clinical characteristics of the involuntary movements in primary and symptomatic dystonias. PATIENTS AND METHODS: 132 consecutive patients with the diagnosis of primary dystonia and 51 consecutive patients with secondary dystonia caused by well defined structural lesion(s) of the central nervous system, with particular emphasis on the characteristics of involuntary movements. RESULTS: Eight variables with the highest risk contribution to either symptomatic or primary dystonias were identified: dystonic movement in secondary dystonia was much more frequently presented at rest, whereas the presence of dystonic tremor, chronic inflammatory process, or peripheral trauma located in the region that is later affected by dystonia, as well as the use of sensory tricks and development of spontaneous remissions, classified the affected patients more often in the category of those with primary dystonia. CONCLUSION: The study identified several clinical features that may be helpful in differentiating primary from secondary dystonia. PMID- 14742623 TI - Cutaneous reactions in patients with solitary cysticercus granuloma on phenytoin sodium. AB - Several medical conditions are believed to be associated with an increased risk of cutaneous adverse reactions to anti-epileptic drugs. The aim of this study was to study the frequency and nature of cutaneous reactions in a cohort of patients being treated with phenytoin sodium for seizures, who were divided into those with a solitary cysticercus granuloma (SCG) and those with a condition other than SCG, to determine if the presence of SCG increases the risk of cutaneous adverse reaction to phenytoin. A cohort of 117, consecutively begun on treatment with phenytoin for seizure control, were followed up prospectively for the development of cutaneous reactions. There were 63 patients with SCG upon imaging and 54 patients to whom phenytoin was administered for seizures due to causes other than SCG or multiple neurocysticercosis. Cutaneous reactions were significantly more common (p = 0.02) in patients with SCG (9/63 patients; 14.3%) than in controls (2/54 patients; 3.7%). The spectrum of skin reactions in patients with SCG included benign skin rash (n = 3), anticonvulsant hypersensitivity syndrome (n = 4), Stevens-Johnson syndrome (n = 1), and urticaria (n = 1). Individuals with seizures due to SCG have a high incidence of cutaneous adverse reactions to phenytoin. This fact should be kept in mind when initiating them on treatment with this anti-epileptic drug. PMID- 14742624 TI - Clinical, molecular, and PET study of a case of aceruloplasminaemia presenting with focal cranial dyskinesia. AB - Aceruloplasminaemia is a rare recessive disorder caused by mutations in the gene encoding the multicopper ferroxidase ceruloplasmin, thought to be involved in cellular iron export. Primary intracellular iron accumulation characterises this disorder. We investigated a case of aceruloplasminaemia early in the course of the disease by structural and functional neuroimaging and correlated the results with the clinical findings. The patient, a diabetic 59 year old lady, presented with perioral dyskinesia. Magnetic resonance imaging (MRI) revealed massive iron accumulation in the basal ganglia, notably sparing the pallidum, and along the cortical surface. However, most of these structures had preserved metabolic activity as evaluated by fluorodeoxyglucose positron emission tomography (FDG PET). Voxel based analysis of FDG-PET data showed a significant hypometabolism only in the heads of the caudate nuclei. Molecular genetic analysis revealed compound heterozygosity for two null mutations in the ceruloplasmin gene, a rather surprising finding for a very rare recessive disease, suggesting that aceruloplasminaemia could be somewhat more frequent than is commonly thought and could therefore be underdiagnosed. PMID- 14742625 TI - A new anti-neuronal antibody in a case of paraneoplastic limbic encephalitis associated with breast cancer. AB - Paraneoplastic neurological syndromes are rare remote effects of different types of cancer. Paraneoplastic limbic encephalitis is a specific syndrome, most often associated with small cell lung carcinoma. This report describes the case of a pure limbic encephalitis in association with breast cancer. An anti-neuronal antibody was found in the serum and CSF of the patient which has not been reported so far. PMID- 14742626 TI - Harlequin syndrome: an association with overlap parasomnia. PMID- 14742627 TI - Tiagabine for treating painful tonic spasms in multiple sclerosis: a pilot study. PMID- 14742628 TI - The validity of using the mini mental state examination in NICE dementia guidelines. PMID- 14742629 TI - An expansion in the ZNF9 gene causes PROMM in a previously described family with an incidental CLCN1 mutation. PMID- 14742630 TI - "When the feeling's gone": a selective loss of musical emotion. PMID- 14742631 TI - Brain tissue guided treatment supplementing ICP/CPP therapy after traumatic brain injury. PMID- 14742632 TI - Dural tear and intracranial hypotension in a chiropractic patient. PMID- 14742633 TI - Potentially misleading extratemporal lobe lesions. PMID- 14742634 TI - Activation of phosphatidylinositol 3-kinase and c-Jun-N-terminal kinase cascades enhances NF-kappaB-dependent gene transcription in BCG-stimulated macrophages through promotion of p65/p300 binding. AB - The proinflammatory response of infected macrophages is an important early host defense mechanism against mycobacterial infection. Mycobacteria have been demonstrated to induce proinflammatory gene transcription through the Toll-like receptors, (TLR)2 and TLR 4, which initiate signaling cascades leading to nuclear factor (NF)-kappaB activation. The main transduction pathway responsible for NF kappaB activation has been established and involves the MyD88, interleukin-1 receptor-associated kinase, tumor necrosis factor receptor-associated factor-6, NF-kappaB-inducing kinase, and inhibitor of kappaB kinase complex. The role of other kinase cascades triggered by mycobacteria in the NF-kappaB activation is less clear. We herein examine the role of the mitogen-activated protein kinases (MAPKs) and phosphatidylinositol 3-kinase (PI-3K) cascades in the expression of the bacillus Calmette-Guerin (BCG) mycobacteria-induced NF-kappaB-dependent genes, macrophage-inflammatory protein-2 (MIP-2) and inducible nitric oxide (NO) synthase. Specific pharmacological inhibition of the PI-3K, c-jun-N-terminal kinase (JNK), and to a smaller extent, p38 MAPK but not extracellular-regulated kinase (ERK), suppressed NF-kappaB-dependent reporter gene transcription and MIP 2 and NO secretion in BCG-induced RAW264.7 macrophages. A similar effect was obtained following molecular inhibition of JNK via JNK-interacting protein-1 overexpression. In addition, a kinase-dead mutant of MEK kinase-1, the up-stream regulator of JNK, also proved to be a potent inhibitor of NF-kappaB-reporter activity. The effect of inhibitors was mediated by the down-regulation of NF kappaB transcription activity and without effecting its nuclear translocation. These data suggest an indirect mechanism of the NF-kappaB regulation by these kinases, probably through p65 phosphorylation and improved binding to the p300 transcription coactivator. The data obtained demonstrate that PI-3K, JNK, and p38 MAPK activation by mycobacteria enhance NF-kappaB-driven gene expression contributing to the proinflammatory macrophage response. PMID- 14742635 TI - Virally stimulated plasmacytoid dendritic cells produce chemokines and induce migration of T and NK cells. AB - The natural interferon (IFN)-producing cell is now known to be identical to the plasmacytoid dendritic cell (PDC). These are Lin-, CD123+, CD11c-, and human leukocyte antigen-DR+ cells that secrete large amounts of IFN-alpha (1-2 IU/cell) when stimulated by enveloped viruses such as herpes simplex virus. In the current study, we have evaluated chemokine expression by virally stimulated PDC. Up regulation of mRNA for CCL4, CCL3, CCL5, CCL2, and CXC chemokine ligand (CXCL)10 in herpes simplex virus-stimulated PDC was detected by RNAse protection assays. In contrast, PDC-depleted peripheral blood mononuclear cells did not up-regulate these mRNA species upon viral stimulation. Enzyme-linked immunosorbent assay and/or intracellular flow cytometry confirmed production of these proteins, and studies indicated overlapping production of IFN-alpha and the other cytokines/chemokines by PDC. Endocytosis plays a critical role in chemokine induction, as disruption of the pathway inhibits the response. However, transcription of viral genes is not required for chemokine induction. Autocrine IFN-alpha signaling in the PDC could account for a portion of the CXCL10 and CCL2 production in virally stimulated PDC but was not responsible for the induction of the other chemokines. To evaluate the functional role of the chemokines, chemotaxis assays were performed using supernatants from virally stimulated PDC. Activated T cells and natural killer cells, but not naive T cells, were preferentially recruited by these PDC supernatants. Migration was subsequently inhibited by addition of neutralizing antibody to CCL4 and CXCL10. We hypothesize that virally induced chemokine production plays a pivotal role in the homing of leukocytes to PDC. PMID- 14742636 TI - Delay of neutrophil apoptosis in acute coronary syndromes. AB - Apoptosis of polymorphonuclear neutrophils (PMN) is currently discussed as a key event in the control of inflammation. This study determined PMN apoptosis and its underlying mechanisms in controls (C), patients with stable (SAP) or unstable angina (UAP), and with acute myocardial infarction (AMI). Blood was drawn from 15 subjects of each C, SAP, UAP, and AMI. Apoptosis was measured by flow cytometry in isolated PMN (propidium iodide staining) and PMN from whole blood (CD16, FcgammaRIII). Serum cytokines were determined by enzyme-linked immunosorbent assay. Apoptosis of isolated PMN was delayed significantly in acute coronary syndromes (ACS) as compared with SAP or C (C, 51.2+/-12.6%; SAP, 44.9+/-13.6%; UAP, 28.4+/-10.1%; AMI, 20.3+/-8.5%; AMI or UAP vs. SAP or C, P<0.001). These results were confirmed by measurement of PMN apoptosis in cultured whole blood from patients and controls. Moreover, serum of patients with ACS markedly reduced apoptosis of PMN from healthy donors. Analysis of patients' sera revealed significantly elevated concentrations of tumor necrosis factor alpha, interferon gamma (IFN-gamma), granulocyte macrophage-colony stimulating factor (GM-CSF), and interleukin (IL)-1beta in ACS (vs. C and SAP). IFN-gamma, GM-CSF, and IL-1beta significantly delayed PMN apoptosis in vitro. Furthermore, coincubation of PMN with adenosine 5'-diphosphate-activated platelets significantly inhibited PMN apoptosis as compared with coculture with unstimulated platelets. This study demonstrates a pronounced delay of PMN apoptosis in UAP and AMI, which may result from increased serum levels of IFN-gamma, GM-CSF, and IL-1beta and from enhanced platelet activation. Therapeutical modulation of these determinants of PMN lifespan may provide a new concept for the control of inflammation in ACS. PMID- 14742637 TI - Diversity in the Sir2 family of protein deacetylases. AB - The silent information regulator (Sir2) family of protein deacetylases (Sirtuins) are nicotinamide adenine dinucleotide (NAD)(+)-dependent enzymes that hydrolyze one molecule of NAD(+) for every lysine residue that is deacetylated. The Sirtuins are phylogenetically conserved in eukaryotes, prokaryotes, and Archeal species. Prokaryotic and Archeal species usually have one or two Sirtuin homologs, whereas eukaryotes typically have multiple versions. The founding member of this protein family is the Sir2 histone deacetylase of Saccharomyces cerevisiae, which is absolutely required for transcriptional silencing in this organism. Sirtuins in other organisms often have nonhistone substrates and in eukaryotes, are not always localized in the nucleus. The diversity of substrates is reflected in the various biological activities that Sirtuins function, including development, metabolism, apoptosis, and heterochromatin formation. This review emphasizes the great diversity in Sirtuin function and highlights its unusual catalytic properties. PMID- 14742638 TI - Soluble Jagged-1 is able to inhibit the function of its multivalent form to induce hematopoietic stem cell self-renewal in a surrogate in vitro assay. AB - Stem cells reside in customized microenvironments (niches) that contribute to their unique ability to divide asymmetrically to give rise to self and to a daughter cell with distinct properties. Notch receptors and their ligands are highly conserved and have been shown to regulate cell-fate decisions in multiple developmental systems through local cell interactions. To assess whether Notch signaling may regulate hematopoiesis to maintain cells in an immature state, we examined the functional role of the recombinant, secreted form of the Notch ligand Jagged-1 during mouse hematopoietic stem cell (HSC) and progenitor cell proliferation and maturation. We found that ligand immobilization on stromal layer or on Sepharose-4B beads is required for the induction of self-renewing divisions of days 28-35 cobblestone area-forming cell. The free, soluble Jagged 1, however, has a dominant-negative effect on self-renewal in the stem-cell compartment. In contrast, free as well as immobilized Jagged-1 promotes growth factor-induced colony formation of committed hematopoietic progenitor cells. Therefore, we propose that differences in Jagged-1 presentation and developmental stage of the Notch receptor-bearing cells influence Notch ligand-binding results toward activation or inhibition of downstream signaling. Moreover, these results suggest potential clinical use of recombinant Notch ligands for expanding human HSC populations in vitro. PMID- 14742639 TI - Preassociation of nonactivated STAT3 molecules demonstrated in living cells using bioluminescence resonance energy transfer: a new model of STAT activation? AB - Signal transducers and activators of transcription (STATs) are crucial molecules in cytokine signaling. In the conventional model of STAT activation, STAT molecules are recruited from a latent pool of cytoplasmic monomers to the activated cytokine receptor. After binding to the receptor, they get tyrosine phosphorylated, dissociate from the receptor, and translocate to the nucleus as activation-induced dimers. Recently, several publications questioned this model of STAT activation and showed the existence of preassociated STAT molecules before activation. We were able to demonstrate the existence of these preassociated STAT3 molecules in living mammalian cells using bioluminescence resonance energy transfer. Our results support the new hypothesis that STAT molecules exist in the cytoplasm as dimers or multimers and point to an activation-induced change in STAT3 conformation. Therefore, we propose a new model of STAT activation and discuss a hypothetical structure of "cytoplasmic" STAT dimers as opposed to the known "activation-induced" dimer. PMID- 14742640 TI - Induction of various immune modulatory molecules in CD34(+) hematopoietic cells. AB - Lipopolysaccharide (LPS) has been shown to induce proliferation of human T lymphocytes only in the presence of monocytes and CD34(+) hematopoietic cells (HCs) from peripheral blood. This finding provided evidence of an active role of CD34(+) HCs during inflammation and immunological events. To investigate mechanisms by which CD34(+) HCs become activated and exert their immune modulatory function, we used the human CD34(+) acute myeloid leukemia cell line KG-1a and CD34(+) bone marrow cells (BMCs). We showed that culture supernatants of LPS-stimulated mononuclear cells (SUP(LPS)) as well as tumor necrosis factor alpha (TauNF-alpha), but not LPS alone, can activate nuclear factor-kappaB in KG 1a cells. By cDNA subtraction and multiplex polymerase chain reaction, we revealed differential expression of cellular inhibitor of apoptosis protein-1, inhibitor of kappaB (IkappaB)/IkappaBalpha (MAD-3), and intercellular adhesion molecule-1 (ICAM-1) in SUP(LPS)-stimulated KG-1a cells and up-regulation of interferon (IFN)-inducible T cell-chemoattractant, interleukin (IL)-8, macrophage inflammatory protein-1alpha (MIP-1alpha), MIP-1beta, RANTES, CD70, granulocyte macrophage-colony stimulating factor, and IL-1beta in stimulated KG-1a cells and CD34(+) BMCs. Although monokine induced by IFN-gamma, IFN-inducible protein 10, and IFN-gamma were exclusively up-regulated in KG-1a cells, differential expression of monocyte chemoattractant protein-1 (MCP-1), macrophage-derived chemokine, myeloid progenitor inhibitory factor-2, and IL-18 receptor was only detectable in CD34(+) BMCs. More importantly, CD34(+) BMCs stimulated by TNF alpha also showed enhanced secretion of MCP-1, MIP-1alpha, MIP-1beta, and IL-8, and increased ICAM-1 protein expression could be detected in stimulated KG-1a cells and CD34(+) BMCs. Furthermore, we revealed that T cell proliferation can be induced by TNF-alpha-stimulated KG-1a cells, which is preventable by blocking anti-ICAM-1 monoclonal antibodies. Our results demonstrate that CD34(+) HCs have the potential to express a variety of immune-regulatory mediators upon stimulation by inflammatory cytokines including TNF-alpha, which may contribute to innate- and adaptive-immune processes. PMID- 14742641 TI - Stimulated human neutrophils form biologically active kinin peptides from high and low molecular weight kininogens. AB - Human neutrophils play a pivotal role in acute inflammation. However, their capacity to generate bioactive kinin peptides has not been established as yet. We have examined the ability of neutrophil enzymes to release biologically active kinins in vitro from purified human H- and L-kininogens. Neutrophils isolated from human blood were stimulated with f-Met-Leu-Phe, thrombin, or human immunoglobulin G adsorbed to silica particles. Supernatants were incubated with iodinated kininogens, and polyacrylamide gel electrophoresis analyzed aliquots taken after a range of incubation times. A time-course analysis demonstrated that supernatants from stimulated neutrophils caused a rapid hydrolysis of both substrates, resulting in an accumulation of fragments ranging from 20 to less than 10 kDa. Radioimmunoassay (RIA) revealed that all supernatants were able to generate kinins in vitro. High-performance liquid chromatography of the generated peptides indicated that they had a retention time similar to that of bradykinin and Met-Lys-bradykinin, clearly recognized as kinin peptides when the corresponding fractions were tested by RIA. The kinin-immunoreactive fractions produced lowering of blood pressure and a dramatic increase in venular permeability. Biological activity of the neutrophil-generated kinins was completely abolished by the B2 receptor antagonist HOE140, indicating that over the time-course of the experiments, only kinin B2 agonists appeared to have been generated and that cellular actions of these were mediated by kinin B2 receptors. Together, our results demonstrate that human neutrophil proteases can release kinins from both plasma kininogens, suggesting that these peptides may participate actively during acute inflammation. PMID- 14742642 TI - A recurring problem: research in restorative dentistry... But there is a light at the end of the tunnel. PMID- 14742643 TI - "Dental Research into Gear", 1970-1998: a review of the scientific legacy of Joop Arends. PMID- 14742644 TI - The molecular basis of salivary gland involvement in graft--vs.--host disease. AB - During the past two decades, the involvement of salivary glands in graft vs. host disease (GVHD) had been intensively researched and published. GVHD occurs in 40 70% of patients treated with bone marrow and peripheral blood stem cell transplantation (PBSCT), and improved survival rates have led to a continuously increasing number of GVHD patients suffering from induced salivary insult. Limited studies suggest that a large percentage of GVHD patients is affected and that the induced salivary dysfunction occurs rapidly following the transplantation. It affects both major and minor salivary glands and reflects the severity of the disease. Moreover, profound sialochemical alterations may be diagnostic of GVHD. An additional reason for this vast research is that GVHD, as an autoimmune-like disease, seemed to be an appropriate model for studying a much more prevalent and well-known and well-studied autoimmune disease involving salivary glands: Sjogren's syndrome. The purpose of the current review-which is, to the best of our knowledge, the first of its kind-is to describe the GVHD related sialometric and sialochemical data published in the past two decades for both major and minor salivary glands and to discuss the pathogenesis and molecular basis of the disease. PMID- 14742645 TI - Blue light differentially modulates cell survival and growth. AB - Previous studies have reported that blue light (400-500 nm) inhibits cell mitochondrial activity. We investigated the hypothesis that cells with high energy consumption are most susceptible to blue-light-induced mitochondrial inhibition. We estimated cell energy consumption by population doubling time, and cell survival and growth by succinate dehydrogenase (SDH) activity. Six cell types were exposed to 5 or 60 J/cm(2) of blue light from quartz-tungsten-halogen (QTH), plasma-arc (PAC), or argon laser sources in monolayer culture. Post-light SDH activity correlated positively with population doubling time (R(2) = 0.91 for PAC, 0.76 for QTH, 0.68 for laser); SDH activity increased for cell types with the longest doubling times and was suppressed for cell types with shorter doubling times. Thus, light-induced exposure differentially affects SDH activity, cell survival, and growth, depending on cell energy consumption. Blue light may be useful as a therapeutic modulator of cell growth and survival. PMID- 14742646 TI - Asymmetry in the condylar long axis and first molar rotation. AB - Asymmetric growth occurs frequently in the mandibulofacial region, but little attention has been given to asymmetry in the temporomandibular joint. The purpose of this study was to clarify the feature of asymmetry in the condylar long axis and its relation to upper first molar rotation. Records of 148 pre-orthodontic patients were used. The angle of the condylar long axis and that of the molar rotation were both larger on the left side than on the right side. Positive correlations were found between the corresponding bilateral measurements of condylar long axes and also between those of molar rotations, whereas no correlation was found between the condylar long axis and molar rotation. These findings were found in most subgroups classified by dental age, skeletal pattern, bite force balance, or gender. These results suggest that consistent left-right differences in the condylar long axis and first molar rotation are common. PMID- 14742647 TI - Antibacterial titanium plate anodized by being discharged in NaCl solution exhibits cell compatibility. AB - Implant surfaces should be modified to achieve excellent cell compatibility as well as antibacterial activity. Our previous study demonstrated that titanium plates anodized by being discharged in NaCl (Ti-Cl) exhibited high antibacterial activity. Since Ti-Cl was prepared with a NaCl solution, we hypothesized that Ti Cl would exhibit low toxicity toward cells. The aims of this study were to characterize the surface of Ti-Cl and investigate the cell compatibility (MC3T3 E1 and L929 cells) of Ti-Cl. The results demonstrated that, since the TiCl(3) formed on the Ti-Cl surface was hydrolyzed into HCl, HClO, and TiOH after immersion in pure distilled water, TiCl(3) contributed to the antibacterial activity of Ti-Cl. On the other hand, TiO formed on the Ti-Cl surface enhanced cell extension and cell growth through a larger adsorption of fibronectin compared with the pure titanium control. These findings suggest that antibacterial titanium is a promising material for use in dental implant systems. PMID- 14742648 TI - Effectiveness of single-surface ART restorations in the permanent dentition: a meta-analysis. AB - Over the past few years, there has been an increase in the number of studies reporting on various aspects of the Atraumatic Restorative Treatment (ART) approach. Five randomized clinical trials in which ART restorations with glass ionomers were compared with amalgam restorations in permanent teeth for a maximum period of 3 yrs constituted the database. This meta-analysis divided the publications into 'early' (1987-1992) and 'late' (1995-) studies on the basis of improvements in the approach. The analysis showed that, in the 'early' studies, single-surface amalgam restorations survived statistically significantly longer than comparable ART restorations after 1, 2, and 3 yrs. This trend did not continue into the late group of studies; no statistically significant difference between the 2 types of restorations was found. Based on the available data, it appears that there is no difference in survival results between single-surface ART restorations and amalgam restorations in permanent teeth over the first 3 yrs. PMID- 14742649 TI - Biting suppresses stress-induced expression of corticotropin-releasing factor (CRF) in the rat hypothalamus. AB - Corticotropin-releasing factor (CRF) expressed in the hypothalamus plays an important role in mediating behavioral responses to stressors. Restraining the body of an animal has been shown to activate and induce an enhanced expression of CRF in paraventricular neurons of the rat hypothalamus. Since aggressive biting behavior is known to suppress stress-induced noradrenaline secretion in the central nervous system and the formation of gastric ulcers, we investigated the effect of biting on restraint-induced CRF expression in the rat hypothalamus. The number of CRF-expressing neurons in the paraventricular nucleus increased significantly after short time restraint (30 or 60 min) followed by a 180-minute post-restraint period. Biting of a wooden stick during the restraint stress significantly suppressed the restraint-induced enhancement of CRF expression in the paraventricular nucleus. These observations suggest a possible anti-stress effect of biting and an important role of para-functional masticatory activity in coping with stressful events. PMID- 14742650 TI - Characterization of dental epithelial progenitor cells derived from cervical-loop epithelium in a rat lower incisor. AB - Dental epithelial progenitor cells differentiate into various cell types during development of tooth germs. To study this mechanism, we produced immortalized dental epithelial progenitor cells derived from the cervical-loop epithelium of a rat lower incisor. The expression patterns of cytokeratin 14, nerve growth factor receptor p75, amelogenin, Notch2, and alkaline phosphatase were examined by immunohistochemistry in both lower and higher cell densities. The patterns of each were compared in the dental epithelium of rat lower incisors. The results demonstrated that these cells could produce ameloblast lineage cells, stratum intermedium cells, stellate reticulum, and outer enamel epithelium. Furthermore, fibroblast growth factor 10 stimulated proliferation of dental progenitor cells and subsequently increased the number of cells expressing alkaline phosphatase. These results suggest that fibroblast growth factor 10 plays a role in coupling mitogenesis of the cervical-loop cells and the production of stratum intermedium cells in rat incisors. PMID- 14742651 TI - In vitro effects of enamel matrix proteins on rat bone marrow cells and gingival fibroblasts. AB - Emdogain (EMD), a formulation of Enamel Matrix Proteins (EMP), is used clinically for periodontal regeneration, where it stimulates cementum formation and promotes gingival healing. In this study, we investigated the in vitro effects of EMD on rat bone marrow stromal cells (BMSC) and gingival fibroblasts (GF). EMD (at 25 micro g/mL) increased the osteogenic capacity of bone marrow, as evidenced by approximately three-fold increase in BMSC cell number and approximately two-fold increase in alkaline phosphatase (ALP) activity and mineralized nodule formation. The presence of EMD in the initial stages (first 48 hrs) of the culture was crucial for this effect. In contrast, EMD did not induce osteoblastic differentiation of GF (evidenced by lack of mineralization or ALP activity) but increased up to two-fold both their number and the amount of matrix produced. These in vitro data on BMSC and GF could explain the promotive effect of EMD on bone formation and connective tissue regeneration, respectively. PMID- 14742652 TI - Lactational exposure of Han/Wistar rats to 2,3,7,8-tetrachlorodibenzo-p-dioxin interferes with enamel maturation and retards dentin mineralization. AB - Exposure to environmental dioxins via mother's milk may be one causative factor of mineralization defects in children's teeth. A prerequisite for the completion of enamel mineralization is the removal of enamel matrix. To test the hypothesis that dioxins interfere with enamel maturation, we administered lactating Han/Wistar rats a single dose of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD; 50 or 1000 micro g/kg) on the day after delivery and analyzed tissue sections of the pup heads at post-natal days (Pn) 9 and 22. By Pn22, the first and second molars of the exposed pups, but not controls, showed retention of enamel matrix. Predentin was thicker than normal. Immunostaining for the aryl hydrocarbon/dioxin receptor (AhR) and cytochrome P4501A1 (CYP1A1) in ameloblasts and odontoblasts was reduced, suggesting that TCDD interferes with tooth mineralization via AhR. Extinction of AhR may lead to abolition of CYP1A1 expression as a sign of impaired dental cell function. PMID- 14742653 TI - Incidence of oral lesions in HIV-1-infected women: reduction with HAART. AB - Few studies assess the effectiveness of HAART on reducing the incidence and recurrence of oral lesions. We investigated such changes among 503 HIV+ women over six years in the Women's Interagency HIV Study. The incidence of erythematous candidiasis (EC), pseudomembranous candidiasis (PC), hairy leukoplakia (HL), and warts was computed over follow-up visits after HAART initiation compared with before HAART initiation. Analysis of our data demonstrates a strong decrease in candidiasis after HAART initiation. The incidence of EC fell to 2.99% from 5.48% (RR 0.545); PC fell to 2.85% from 6.70% (RR 0.425); and EC or PC fell to 3.43% from 7.35% (RR 0.466). No changes were seen in HL or warts. Higher HIV-RNA was associated with greater incidence of candidiasis and HL, but not warts. Analysis of these data indicates that recurrence and incidence of candidiasis are reduced by HAART, and that recurrence is reduced independently of CD4 and HIV-RNA. PMID- 14742654 TI - Periodontal disease and biomarkers related to cardiovascular disease. AB - Periodontal disease is a chronic infection of the gums characterized by a loss of attachment between the tooth and bone, and by bone loss. We evaluated cross sectionally the association between periodontal disease and C-reactive protein (CRP), fibrinogen, factor VII, tissue plasminogen activator (t-PA), LDL-C, von Willebrand factor, and soluble tumor necrosis factor receptors 1 and 2. The final sample consisted of 468 men (ages 47-80 yrs), participating in the Health Professional Follow-up Study, who provided blood and were free of CVD, diabetes, and cancer. In multivariate regression models controlling for age, cigarette smoking, alcohol intake, physical activity, and aspirin intake, self-reported periodontal disease was associated with significantly higher levels of CRP (30% higher among periodontal cases compared with non-cases), t-PA (11% higher), and LDL-C (11% higher). Based on our data, periodontal disease showed significant associations with biomarkers of endothelial dysfunction and dyslipidemia, which may potentially mediate the association between periodontal and cardiovascular disease. PMID- 14742655 TI - Periodontitis and systemic inflammation: control of the local infection is associated with a reduction in serum inflammatory markers. AB - Severe periodontitis is associated with elevated inflammatory markers in otherwise healthy populations. However, the nature of this association has not been determined. Our aim was to assess whether the degree of response to periodontal therapy was associated with changes in serological markers of systemic inflammation. Ninety-four systemically healthy subjects with severe generalized periodontitis participated in a prospective six-month blind intervention trial. Periodontal parameters and inflammatory markers [C-reactive Protein (CRP) and Interleukin-6 (IL-6)] were evaluated prior to and 2 and 6 mos after delivery of standard non-surgical periodontal therapy. Six months after treatment, significant reductions in serum IL-6 (p < 0.001, median decrease 0.2 ng/L, 95% CI 0.1-0.4 ng/L) and CRP (p < 0.0001, median decrease 0.5 mg/L, 95% CI 0.4-0.7) were observed. Decreases in inflammatory markers were significant in subjects with above average clinical response to periodontal therapy after correction for possible confounders. Periodontitis may add to the systemic inflammatory burden of affected individuals. PMID- 14742656 TI - Association of ALDH(2) genotypes and alcohol consumption with periodontitis. AB - There is little information regarding the association between alcohol consumption and periodontitis risk. We assessed whether alcohol consumption and ALDH(2) genotypes were associated with periodontitis. Subjects' lifestyle was examined by a self-administered questionnaire, and the percentage of pocket depths > or = 3.5 mm was used as a periodontal parameter. ALDH(2) genotypes were determined with the use of a PCR/RFLP method. Multiple logistic analyses showed that alcohol consumption was significantly associated with periodontitis, and its odds ratio was 1.98. There was no significant relationship between periodontal status and ALDH(2) genotypes. However, ALDH(2)*1/*2 subjects who consumed > or = 33 g/day of alcohol had a significantly greater percentage of pocket depths > or = 3.5 mm than those whose daily consumption was lower, while there was no significant difference in periodontal status associated with alcohol consumption in ALDH(2)*1/*1 subjects. Our results suggest that alcohol consumption may be a risk indicator for periodontitis in ALDH(2)*1/*2 subjects who consume larger amounts of alcohol. PMID- 14742657 TI - Differential expression of RANKL and osteoprotegerin in gingival crevicular fluid of patients with periodontitis. AB - The receptor activator for NF-kappaB ligand (RANKL) plays an important role in osteoclast formation. A recent study with animal models suggests the involvement of RANKL in the pathogenesis of this periodontal disease. However, no one has examined the level of RANKL in the body fluid of human subjects. This communication reports on the in vivo concentrations of RANKL and the RANKL decoy receptor osteoprotegerin (OPG) in the gingival crevicular fluid (GCF) of periodontal subjects with severe, moderate, and mild forms of the disease. An increased concentration of RANKL and a decreased concentration of OPG were detected in GCF from patients with periodontitis (*p < 0.05 vs. control subjects). The ratio of the concentration of RANKL to that of OPG in the GCF was significantly higher for periodontal disease patients than for healthy subjects (*p < 0.01). Taken together, these data suggest that RANKL and OPG contribute to osteoclastic bone destruction in periodontal disease. ABBREVIATIONS: GCF, gingival crevicular fluid; IL, interleukin; OPG, osteoprotegerin; RANKL, receptor activator for NF-kappaB ligand. PMID- 14742658 TI - Bacteremia following surgical dental extraction with an emphasis on anaerobic strains. AB - Our aim was to investigate bacteremia caused by surgical extraction of partly erupted mandibular third molars. From 16 young adults, bacterial samples were taken from the third-molar pericoronal pocket and post-operatively from the extraction socket, and blood samples were drawn from the ante-cubital vein up to 30 min after surgery. Of the subjects, 88% had detectable bacteremia-50% 1 min after the incision, 44% immediately after extraction. The respective percentages at 10, 15, and 30 min were 44%, 25%, and 13%. Blood cultures contained 31 species (74% anaerobes), with 3.9 +/- 2.6 species isolated per subject. Most prevalent were the anaerobes Prevotella, Eubacterium, and Peptostreptococcus sp. and the aerobes viridans-group streptococci and Streptococcus milleri group. Any species found in the blood was also isolated from the mouth, from 93% of the pericoronal pockets and from 43% of the extraction sockets. Surgical dental extraction clearly causes bacteremia of a high frequency and lasting longer than thus far assumed. PMID- 14742659 TI - Dental caries, oral hygiene, and oral clearance in children with craniofacial disorders. AB - The reason that children with cleft palates tend to have a greater prevalence of tooth decay than normal children is unclear. We hypothesized that children with cleft palates would have increased oral clearance times for foods and, consequently, higher levels of caries and caries-associated micro-organisms than control children. Children aged 6-16 yrs, with (n = 81) or without (n = 61) cleft palates, were studied. Children with cleft palates had DMFT and dmft scores greater (p < 0.01) than those of the control group. The number of caries associated organisms was greater in the saliva of the cleft palate children (all p < 0.001). The oral hygiene, plaque and gingival index scores were greater (p < 0.0001), oral clearance was longer (p < 0.01), and levels of sucrose and starch derived saccharides higher (p < 0.01) in the cleft palate group. However, salivary concentrations of organic acids were lower in the children with craniofacial disorders, probably reflecting the altered physiology of the more mature dental biofilm. The longer oral clearance times of foods and the consequent generation of fermentable sugars from starches may contribute to the higher caries prevalence observed in children with cleft palates. PMID- 14742660 TI - Effect of acculturation on objective measures of oral health in Haitian immigrants in New York City. AB - Acculturation is a complex phenomenon that can serve as a proxy for cultural norms and behaviors affecting care-seeking, prevention behaviors, and, ultimately, health outcomes. The purpose of this study was to assess the effect of acculturation on the oral health of Haitian immigrants in New York City. We hypothesized that acculturation would be a predictor of the oral health status of the participating individuals. An acculturation scale was specifically developed and validated for this study. A sample of 425 adult Haitian immigrants living in NYC was obtained through outreach activities. Oral health examinations were conducted, and a questionnaire was administered to the participants. After adjustment for age, sex, education, income, and marital status, acculturation was negatively associated with measures of decayed teeth, periodontal attachment loss of > or = 4 mm, and the number of missing teeth. Results suggest a positive impact of acculturation on the oral health status of these individuals. PMID- 14742661 TI - Changes in masseteric hemodynamics time-related to mental stress. AB - Mental stress may cause a dissociation of sympathetic outflow to different regions. However, it remains unclear how the sympathetic outflow to jaw muscles is related to other sympathetic outflow under mental stress. The objective of this study was to clarify the temporal relationship between the finger sweat expulsion elicited by mental stress and the hemodynamic and electromyographic changes in the masseter muscle. Healthy adult female volunteers participated in this study. Masseteric hemodynamic changes were closely time-related to mental stress, showing a decrease in oxygen saturation of muscle blood around the onset of mental stress. In contrast, EMG activity of jaw-closing muscles was not time related to mental stress. These results suggest that mental stress induces hemodynamic changes that are not associated with EMG activity in the masseter muscle of healthy adult females. PMID- 14742662 TI - Identification and characterization of two forms of mouse MUTYH proteins encoded by alternatively spliced transcripts. AB - There are three types of mouse Mutyh mRNAs (type a, b and c) generated by alternative splicing, and type b mRNA is a major form among the three in most of the tissues examined. The level of type c mRNA is relatively high in brain. Type a and b mRNAs were expected to encode 57.7 kDa protein (MUTYHalpha), while type c mRNA had a partly different open reading frame encoding a 50.2 kDa protein (MUTYHbeta). An in vitro translation of type b and c mRNAs produced a 50 kDa MUTYHalpha and 47 kDa MUTYHbeta, respectively. MUTYHalpha and MUTYHbeta were detected in wild-type embryonic stem (ES) cells or thymocytes prepared from wild type mice, but neither MUTYH-null ES cells nor thymocytes prepared from MUTYH null mice. Both MUTYHalpha and MUTYHbeta were mainly localized in the nuclei and some in mitochondria in wild-type ES cells. Recombinant MUTYHalpha and beta were expressed as fusion proteins with thioredoxin in Escherichia coli, but only MUTYHalpha was partly soluble and thus could be purified. Recombinant MUTYHalpha possessed DNA glycosylase activities to excise adenine opposite 8-oxoguanine and guanine but not AP lyase activity. PMID- 14742663 TI - Premature termination codons enhance mRNA decapping in human cells. AB - Nonsense-mediated mRNA decay (NMD) is a eukaryotic surveillance process that promotes selective degradation of imperfect messages containing premature translation termination codons (PTCs). In yeast, PTCs trigger both deadenylylation-independent mRNA decapping, thereby allowing their rapid degradation by a 5' to 3' exonuclease, and to a smaller extent accelerated deadenylylation. It is not clear to what extent this decay pathway is conserved in higher eukaryotes. We used a transcriptional pulse strategy relying on a tetracycline-regulated promoter to study the decay of a PTC- containing beta globin mRNA in human cells. We show that a PTC destabilizes the mRNA and decreases its half-life from >16 h to 3 h. The deadenylylation rate is increased, but not sufficiently to account for the decreased half-life on its own. Using a circularization RT-PCR (cRT-PCR) strategy, we could detect decapped degradation intermediates and measure simultaneously their poly(A) tail length. This allowed us to show that a PTC enhances the rate of mRNA decapping and that decapped products have been deadenylylated to a certain extent. Thus the major feature of the NMD pathway, enhanced decapping, is conserved from yeast to man even though the kinetic details might differ between various mRNAs and/or species. PMID- 14742664 TI - Synthesis of phosphorothioamidites derived from 3'-thio-3'-deoxythymidine and 3' thio-2',3'-dideoxycytidine and the automated synthesis of oligodeoxynucleotides containing a 3'-S-phosphorothiolate linkage. AB - The synthesis of N4-benzoyl-5'-O-dimethoxytrityl-2',3'-dideoxy-3'-thiocytidine and its phosphorothioamidite is described for the first time, together with a shortened procedure for the preparation of 5'-O-dimethoxytrityl-3'-deoxy-3' thiothymidine and its corresponding phosphorothioamidite. The first fully automated coupling procedure for the incorporation of a phosphorothioamidite into a synthetic oligodeoxynucleotide has been developed, which conveniently uses routine activators and reagents. Coupling yields using this protocol were in the range of 85-90% and good yields of singularly modified oligonucleotides were obtained. Coupling yields were also equally good when performed on either a 0.2 or 1 micro mol reaction column, thus facilitating large scale syntheses required for mechanistic studies. PMID- 14742665 TI - More active human L1 retrotransposons produce longer insertions. AB - The vast majority of L1 insertions are 5' truncated and thus inactive. Yet, the mechanism of 5' truncation is unknown. To examine whether the frequency of L1 retrotransposition is directly correlated with the length of genomic L1 insertions, we used a cell culture assay to measure retrotransposition frequency and a PCR-based assay to measure L1 insertion length. We tested five full-length human L1 elements that retrotranspose at different frequencies: LRE3, L1(RP), L1.3, L1.2A and L1.2B. Our data suggest that L1 insertion length correlates with L1 retrotransposition frequency for insertions >1 kb in length. For two elements, L1(RP) and L1.2A, we found that swapping the reverse transcriptase domains had little effect. Instead, we found that genomic insertion length and retrotransposition frequency are substantially affected by amino acid substitutions at positions 363, 1220 and 1259 in ORF2. We suggest that the region containing residues 1220 and 1259 may be important in the binding of ORF2p to L1 RNA to facilitate reverse transcription. PMID- 14742667 TI - Properties and functions of GAF domains in cyclic nucleotide phosphodiesterases and other proteins. PMID- 14742666 TI - Interaction of Saccharomyces Cdc13p with Pol1p, Imp4p, Sir4p and Zds2p is involved in telomere replication, telomere maintenance and cell growth control. AB - Telomeres are the physical ends of eukaryotic chromosomes. They are important for maintaining the integrity of chromosomes and this function is mediated through a number of protein factors. In Saccharomyces cerevisiae, Cdc13p binds to telomeres and affects telomere maintenance, telomere position effects and cell cycle progression through G(2)/M phase. We identified four genes encoding Pol1p, Sir4p, Zds2p and Imp4p that interact with amino acids 1-252 of Cdc13p using a yeast two hybrid screening system. Interactions of these four proteins with Cdc13p were through direct protein-protein interactions as judged by in vitro pull-down assays. Direct protein-protein interactions were also observed between Pol1p Imp4p, Pol1p-Sir4p and Sir4p-Zds2p, whereas no interaction was detected between Imp4p-Sir4p and Zds2p-Imp4p, suggesting that protein interactions were specific in the complex. Pol1p was shown to interact with Cdc13p. Here we show that Zds2p and Imp4p also form a stable complex with Cdc13p in yeast cells, because Zds2p and Imp4p co-immunoprecipitate with Cdc13p, whereas Sir4p does not. The function of the N-terminal 1-252 region of Cdc13p was also analyzed. Expressing Cdc13(252 924)p, which lacks amino acids 1-252 of Cdc13p, causes defects in progressive cell growth and eventually arrested in the G(2)/M phase of the cell cycle. These growth defects were not caused by progressive shortening of telomeres because telomeres in these cells were long. Point mutants in the amino acids 1-252 region of Cdc13p that reduced the interaction between Cdc13p and its binding proteins resulted in varying level of defects in cell growth and telomeres. These results indicate that the interactions between Cdc13(1-252)p and its binding proteins are important for the function of Cdc13p in telomere regulation and cell growth. Together, our results provide evidence for the formation of a Cdc13p-mediated telosome complex through its N-terminal region that is involved in telomere maintenance, telomere length regulation and cell growth control. PMID- 14742668 TI - Lessons from the CYP3A4 promoter. PMID- 14742669 TI - Design of a high-affinity competitive antagonist of the vanilloid receptor selective for the calcium entry-linked receptor population. AB - We describe the synthesis and characterization of N-(4-chlorobenzyl) -N'-(4 hydroxy-3-iodo-5-methoxybenzyl)thiourea (IBTU), a novel antagonist of the vanilloid receptor 1 (TRPV1 or VR1). IBTU competitively inhibited 45Ca2+ uptake into CHO cells heterologously expressing rat TRPV1, whether induced by capsaicin or resiniferatoxin (Ki = 99 +/- 23 and 93 +/- 34 nM, respectively). IBTU was thus somewhat more potent (5-fold) than capsazepine. In contrast to its antagonism of vanilloid-induced calcium uptake, IBTU (30 microM) inhibited [3H]resiniferatoxin binding to TRPV1 by less than 10%. We hypothesize that these dramatically distinct potencies reflect different fractions of TRPV1 in this system: namely, a minor plasma membrane fraction controlling 45Ca2+ uptake, and the predominant intracellular fraction that dominates the [3H]resiniferatoxin binding measurements. Intracellular Ca2+ imaging supports this explanation. IBTU antagonized the elevation in intracellular Ca2+ in response to 50 nM capsaicin with an IC50 of 106 +/- 35 nM. Likewise, 600 nM IBTU was able to antagonize the elevation in intracellular Ca2+ in response to 100 pM resiniferatoxin in the presence of normal (1.8 mM) extracellular Ca2+, where the increase in intracellular calcium reflects calcium influx. In contrast, in the absence of extracellular Ca2+, where in this system resiniferatoxin induces a modest increase in calcium from intracellular stores, IBTU was unable to block the response to resiniferatoxin, although the TRPV1 antagonist 5-iodoresiniferatoxin was able to do so. In summary, IBTU is a novel, potent TRPV1 antagonist with marked selectivity between subpopulations of TRPV1 and may permit the function of these distinct pools to be explored and potentially exploited. PMID- 14742670 TI - A novel constitutive androstane receptor-mediated and CYP3A-independent pathway of bile acid detoxification. AB - Cytosolic sulfotransferase (SULT)-mediated sulfation plays an essential role in the detoxification of bile acids and is necessary to avoid pathological conditions, such as cholestasis, liver damage, and colon cancer. In this study, using transgenic mice bearing conditional expression of the activated constitutive androstane receptor (CAR), we demonstrate that activation of CAR is both necessary and sufficient to confer resistance to the hepatotoxicity of lithocholic acid (LCA). Surprisingly, the CAR-mediated protection is not attributable to the expected and previously characterized CYP3A pathway; rather, it is associated with a robust induction of SULT gene expression and increased LCA sulfation. We have also provided direct evidence that CAR regulates SULT expression by binding to the CAR response elements found within the SULT gene promoters. Interestingly, activation of CAR was also associated with an increased expression of the 3'-phosphoadenosine 5'-phosphosulfate synthetase 2 (PAPSS2), an enzyme responsible for generating the sulfate donor 3'-phosphoadenosine-5' phosphosulfate. Analysis of gene knockout mice revealed that CAR is also indispensable for ligand-dependent activation of SULT and PAPSS2 in vivo. Therefore, we establish an essential and unique role of CAR in controlling the mammalian sulfation system and its implication in the detoxification of bile acids. PMID- 14742671 TI - Multiple pharmacophores for the investigation of human UDP glucuronosyltransferase isoform substrate selectivity. AB - The UDP-glucuronosyltransferase (UGT) enzyme 'superfamily' contributes to the metabolism of a myriad of drugs, nondrug xenobiotic agents, and endogenous compounds. Although the individual UGT isoforms exhibit distinct but overlapping substrate selectivities, structural features of substrates that confer selectivity remain largely unknown. Using methods developed for pharmacophore fingerprinting combined with optimization and pattern recognition techniques, subsets of pharmacophores associated with the substrates and nonsubstrates of 12 human UGT isoforms were selected to generate predictive models of substrate selectivity and to elucidate the chemical and structural features associated with substrates and nonsubstrates. For all 12 UGT isoforms, the pharmacophore model generated showed predictive ability, as determined by a test set comprising 30% of the available data for each isoform. Models for UGT1A6, -1A7, -1A9, and -2B4 displayed the best predictive ability (>75% of test set predicted correctly) and were further analyzed to interpret the pharmacophores selected as important. The individual pharmacophores differed among isoforms but generally represented relatively simple structural and chemical features. For example, an aromatic ring attached to the nucleophilic group was found to increase the likelihood of glucuronidation by UGT1A6, UGT1A7 and UGT1A9. A large hydrophobic region close to the nucleophile and a hydrogen bond acceptor 10 A from the nucleophile were found to be common to most UGT2B4 substrates. The pharmacophores further suggest that the environment immediately adjacent to the nucleophilic site of conjugation is an important determinant of metabolism by a particular UGT. PMID- 14742672 TI - Peroxisome proliferator-activated receptor-gamma-independent repression of collagenase gene expression by 2-cyano-3,12-dioxooleana-1,9-dien-28-oic acid and prostaglandin 15-deoxy-delta(12,14) J2: a role for Smad signaling. AB - Matrix metalloproteinases (MMPs) degrade extracellular matrix components, and overexpression of these enzymes contributes to tissue destruction in arthritis. Of particular importance are the collagenases, MMP-1 and MMP-13, which have high activity against the interstitial collagens in cartilage. In this study, we address the mechanisms of two inhibitors of collagenase gene expression, the synthetic triterpenoid 2-cyano-3,12-dioxooleana-1,9-dien-28-oic acid (CDDO) and 15-deoxy-delta(12,14)-prostaglandin J2 (15-dPGJ2). Although both inhibitors are ligands for the nuclear hormone receptor peroxisome proliferator-activated receptor-gamma (PPAR-gamma), a connection between PPAR-gamma and collagenase gene expression has yet to be established. Here, we test the hypothesis that CDDO and 15-dPGJ2 use PPAR-gamma to repress MMP gene expression. Our findings with the PPAR-gamma antagonist 2-[4-[2-[3-(2,4-difluorophenyl)-1-heptylureido]ethyl]rsqb] phenylsulfanyl]-2-methylpropionic acid (GW9662) and mouse embryonic fibroblasts lacking PPAR-gamma demonstrate that CDDO and 15-dPGJ2 use PPAR-gamma-independent mechanisms to inhibit collagenase gene expression. To address a potential PPAR gamma-independent mechanism leading to the repression of MMPs by CDDO, we tested the effect of CDDO on the transforming growth factor-beta (TGF-beta) signaling pathway. We found that CDDO requires Smads (transcription factors activated by TGF-beta) for the repression of MMP-1. Specifically, MMP-1 is inhibited neither by CDDO in the absence of TGF-beta receptor-activated Smad3 nor when a negative regulator, Smad7, attenuates TGF-beta signaling. We conclude that CDDO represses MMP gene expression through a novel PPAR-gamma-independent mechanism that requires Smad signaling. PMID- 14742673 TI - Nitric oxide regulates c-fos expression in nucleus tractus solitarii induced by baroreceptor activation via cGMP-dependent protein kinase and cAMP response element-binding protein phosphorylation. AB - Activation of the arterial baroreceptors induces expression of the proto-oncogene c-fos in the nucleus tractus solitarii (NTS), the terminal site of baroreceptor afferents in the medulla oblongata. This induced expression is an intracellular event that is crucial to long-term maintenance of stable blood pressure. Using Sprague-Dawley rats maintained under propofol anesthesia, we evaluated the role and delineated the underlying molecular mechanisms of nitric oxide (NO) in this process. Baroreceptor activation induced by 30 min of sustained hypertension significantly and sequentially increased the level of cyclic GMP-dependent protein kinase I (PKG-I), phosphorylated cyclic AMP response element-binding protein (pCREB), c-fos mRNA, and Fos protein in the NTS. All of these up regulated expressions were significantly attenuated in animals that were pretreated immediately before baroreceptor activation with bilateral microinjection into the NTS of a selective neuronal nitric-oxide synthase (nNOS) inhibitor, 7-nitroindazole (2.5 pmol), or a soluble guanylyl cyclase (sGC) inhibitor, 1-H-[1,2,4]oxadiaolo[4,3-a]quinoxalin-1-one (1 nmol). Bilateral NTS microinjection of a cell-permeable cGMP analog, 8-bromoguanosine-3',5'-cyclic monophosphate (10 nmol) significantly elevated the level of pCREB or c-fos mRNA in the NTS. On the other hand, the up-regulated CREB phosphorylation or c-fos induction evoked in the dorsomedial medulla by baroreceptor activation was significantly antagonized by NTS application of a cell-permeable cGMP antagonist, (R)p-8-bromoguanosine-3',5'-cyclic monophosphorothioate (5 nmol), or a PKG inhibitor, (8R,9S,11S)-(-)-9-methoxy-carbamyl-8-methyl-2,3,9,10-tetrahydro-8,11 epoxy-1H,8H,11H,-2,7b,11a-trizadizo-benzo(a,g)cycloocta(c,d,e)-trinden-1-one (1 nmol). We conclude that NO derived from nNOS in the NTS on baroreceptor activation may participate in c-fos expression via phosphorylation of CREB in a process that engages the sGC/cGMP/PKG-I signaling cascade. PMID- 14742674 TI - Identification of a novel polymorphic enhancer of the human CYP3A4 gene. AB - CYP3A4, the most abundant form of cytochrome P450 in the human adult liver, shows wide interindividual variation in its activity. This variability is thought to be caused largely by transcriptional and genetic factors, yet the underlying mechanisms are poorly understood. The purpose of this study was to clarify the mechanisms controlling the CYP3A4 gene transcription and to search for genetic polymorphisms in the 5'-flanking region of the CYP3A4 gene. Transient transfection of human hepatoma HepG2 cells and of normal human hepatocytes with a series of CYP3A4 promoter-luciferase reporter plasmids revealed that a region from -11.4 to -10.5 kilobases, designated the constitutive liver enhancer module of CYP3A4 (CLEM4), was important for the constitutive activation of the CYP3A4 gene. Gel shift assay using nuclear extracts prepared from HepG2 cells showed that HNF-1alpha, HNF-4alpha, USF1, and AP-1 interacted with CLEM4. Furthermore, the introduction of mutations into their binding sites demonstrated that essentially all sites were required for the maximal enhancer activity. Screening for genetic polymorphisms within CLEM4 in genomic DNA from French persons, we identified the novel variant, TGT insertion between -11,129 and -11,128 (-11,129_ 11,128insTGT), whose allele frequency was 3.1%. The -11,129_-11,128insTGT resulted in the disruption of USF1 binding and a 36% reduction of the enhancer activity. These results suggest that CLEM4 is a constitutive enhancer of the CYP3A4 gene in the liver and that -11,129_-11,128insTGT may at least partly contribute to the interindividual variability of CYP3A4 expression. PMID- 14742675 TI - New molecular determinants controlling the accessibility of ouabain to its binding site in human Na,K-ATPase alpha isoforms. AB - Inhibition of Na,K-ATPase alpha2 isoforms in the human heart is supposed to be involved in the inotropic effect of cardiac glycosides, whereas inhibition of alpha1 isoforms may be responsible for their toxic effects. Human Na,K-ATPase alpha1 and alpha2 isoforms exhibit a high ouabain affinity but significantly differ in the ouabain association and dissociation rates. To identify the structural determinants that are involved in these differences, we have prepared chimeras between human alpha1 and alpha2 isoforms and alpha2 mutants in which nonconserved amino acids were exchanged with those of the alpha1 isoform, expressed these constructs in Xenopus laevis oocytes, and measured their ouabain binding kinetics. Our results show that replacement of Met119 and Ser124 in the M1-M2 extracellular loop of the alpha2 isoform by the corresponding Thr119 and Gln124 of the alpha1 isoform shifts both the fast ouabain association and dissociation rates of the alpha2 isoform to the slow ouabain binding kinetics of the alpha1 isoform. The amino acids at position 119 and 124 cooperate with the M7 M8 hairpin and are also responsible for the small differences in the ouabain affinity of the ouabain-sensitive alpha1 and alpha2 isoforms. Thus, we have identified new structural determinants in the Na,K-ATPase alpha-subunit that are involved in ouabain binding and probably control, in an alpha isoform-specific way, the access and release of ouabain to and from its binding site. PMID- 14742676 TI - Proteasome inhibitors induce inhibitory kappa B (I kappa B) kinase activation, I kappa B alpha degradation, and nuclear factor kappa B activation in HT-29 cells. AB - The transcription factor nuclear factor kappaB (NF-kappaB) is activated and seems to promote oncogenesis in certain cancers. A major mechanism of NF-kappaB activation in cells involves cytoplasm-to-nucleus translocation of this transcription factor after hydrolysis of the cytoplasmic inhibitor inhibitory kappaB (IkappaB) by the 26S proteasome. Because selective proteasome inhibitors have been shown to block IkappaB degradation; consequently, NF-kappaB activation in a variety of cellular systems, proteasome inhibitors were proposed as potential therapeutic agents for the treatment of cancer. However, under certain conditions, IkappaB degradation and NF-kappaB activation are not mediated by the proteasome system. We investigated how proteasome inhibitors affected NF-kappaB activation in the intestinal epithelial cancer cell line HT-29, which has been documented to have an atypical NF-kappaB regulation. Treatment of cells with the selective proteasome inhibitors carbobenzoxy-L-leucyl-L-leucyl-L-norvalinal (MG 115), carbobenzoxy-L-leucyl-L-leucyl-L-leucinal (MG-132), or lactacystin induced NF-kappaB activation as indicated by both an increase in NF-kappaB DNA binding and transcriptional activity. This increase in NF-kappaB activation caused by proteasome inhibitors was accompanied by an increase in IkappaB kinase activation and a degradation of IkappaBalpha but not IkappaBbeta. Furthermore, proteasome inhibitors induced the expression of NF-kappaB target genes. In summary, these results demonstrate a unique effect of proteasome inhibitors on the IkappaB-NF kappaB systems in HT-29 cells, in which proteasome inhibitors activate rather than deactivate the NF-kappaB system. We conclude that the use of proteasome inhibitors to block NF-kappaB activation in cancer cells may not always be a viable approach. PMID- 14742677 TI - Barbiturates directly inhibit the calmodulin/calcineurin complex: a novel mechanism of inhibition of nuclear factor of activated T cells. AB - Barbiturates are frequently used for the treatment of intracranial hypertension after brain injury but their application is associated with a profound increase in the infection rate. The mechanism of barbiturate-induced failure of protective immunity is still unknown. We provide evidence that nuclear factor of activated T cells (NFAT), an essential transcription factor in T cell activation, is a target of barbiturate-mediated immunosuppression in human T lymphocytes. Treatment of primary CD3+ lymphocytes with barbiturates inhibited the PMA and ionomycin induced increase in DNA binding of NFAT, whereas the activity of other transcription factors, such as Oct-1, SP-1, or the cAMP response element-binding protein, remained unaffected. Moreover, barbiturates suppressed the expression of a luciferase reporter gene under control of NFAT (stably transfected Jurkat T cells), and of the cytokine genes interleukin-2 and interferon-gamma that contain functional binding motifs for NFAT within their regulatory promotor domains (human peripheral blood CD3+ lymphocytes). Neither GABA receptor-initiated signaling nor direct interactions of barbiturates with nuclear proteins affected the activity of NFAT. In contrast, barbiturates suppressed the calcineurin dependent dephosphorylation of NFAT in intact T cells and also inhibited the enzymatic activity of calcineurin in a cell-free system, excluding upstream regulation. Thus, our results demonstrate a novel mechanism of direct inhibition of the calcineurin/calmodulin complex that may explain some of the known immunosuppressive effects associated with barbiturate treatment. PMID- 14742678 TI - The type 1 angiotensin II receptor tail affects receptor targeting, internalization, and membrane fusion properties. AB - Endocytosis modulates cell responses by removing and recycling receptors from the cell surface. Type I angiotensin II receptors (AT1R) are somewhat unique in that they are expressed at apical (AP) and basolateral (BL) membranes in proximal tubule cells and both receptor sites undergo endocytosis. We analyzed AT1R cytoplasmic (-COOH) tail deletion mutants to determine whether classic AT1R endocytosis motifs functioned similarly in polarized cells and simultaneously altered receptor properties. Serially truncating the AT1R tail had little effect on AP/BL AT1R distribution as determined by 125I-angiotensin II binding in LLCPK(Cl4) cells transfected with an AT1R transcript. AP AT1R expression required the proximal 12 amino acids in the AT1R-COOH tail. Deleting all but the proximal 12 aa of the AT1R-COOH tail (T316L mutant) decreased AP AT1R internalization at 20 min (17 +/- 6%; p < 0.05 versus full-length; n = 5) and inhibited AP AT1R stimulated arachidonic acid release (counts released per milligram of protein at 20 min: full-length, 18,762 +/- 4018; T316L, 2430 +/- 1711; n = 4; p < 0.02). Endosomal fusion assays were performed using peptide sequences of regions in the AT1R tail involved in endocytosis (YFLQLLKYIPP [LL] and LSTKMSTLSY [STL]). Peptide STL significantly inhibited endosomal fusion (22 +/- 10% of control; n = 5; p < 0.05 versus positive control). Peptide LL had no significant inhibitory effect. AT(1)R in polarized cells contain dominant endocytosis signals but these motifs do not correlate with AP or BL AT1R expression. Moreover, peptide sequences within the AT1R-COOH tail necessary for endocytosis also modulate endosomal fusion properties. PMID- 14742679 TI - Use-dependent inhibition of the N-methyl-D-aspartate currents by felbamate: a gating modifier with selective binding to the desensitized channels. AB - Felbamate (FBM) is a potent nonsedative anticonvulsant whose clinical effect may be related to the inhibition of N-methyl-D-aspartate (NMDA) currents, but the exact molecular action remains unclear. Using whole-cell patch-clamp recording in rat hippocampal neurons, we found that submillimolar FBM effectively modifies the gating process of NMDA channels. During a single high-concentration (1 mM) NMDA pulse, FBM significantly inhibits the late sustained current but not the early peak current. However, if the 1 mM NMDA pulse is preceded by a low-concentration (10 microM) NMDA prepulse, then FBM significantly inhibits both the peak and the sustained currents in the 1 mM pulse. In sharp contrast, the NMDA currents elicited by micromolar NMDA are only negligibly inhibited or even enhanced by FBM. These findings indicate that the inhibitory effect of FBM on NMDA currents is stronger with both higher NMDA concentration and longer NMDA exposure, and is thus "use-dependent". FBM also slows recovery of the desensitized NMDA channel, and quantitative analyses of FBM effects on the activation kinetics and the desensitization curve of the NMDA currents further disclose dissociation constants of approximately 200, approximately 110, and approximately 55 microM for FBM binding to the resting, activated, and desensitized NMDA channels, respectively. We conclude that therapeutic concentrations (50-300 microM) of FBM could bind to and modify a significant proportion of the resting NMDA channel even when NMDA or other glutamatergic ligand is not present and then decrease the NMDA currents at subsequent NMDA pulses by stabilization of the desensitized channels. Because the inhibitory effect is apparent only when there is excessive NMDA exposure, FBM may effectively inhibit many seizure discharges but preserve most normal neuronal firings. PMID- 14742680 TI - Differential modulation of CaV2.3 Ca2+ channels by Galphaq/11-coupled muscarinic receptors. AB - CaV2.3 subunits are expressed in neuronal and neuroendocrine cells where they are believed to form native R-type Ca2+ channels. Although R-type currents are involved in triggering neurotransmitter and hormone secretion, little is known about their modulation. Previous studies have shown that muscarinic acetylcholine receptors evoke both inhibition and stimulation of CaV2.3. Muscarinic inhibition of CaV2.3 is mediated by Gbetagamma subunits, whereas stimulation is mediated by pertussis toxin-insensitive Galpha subunits. In the present study, we compared modulation of CaV2.3 by the three Galphaq/11-coupled muscarinic receptors (M1, M3, and M5). Our data indicate that these receptors trigger comparable stimulation of CaV2.3. The signaling pathway that mediates stimulation was meticulously analyzed for M1 receptors. Stimulation is blocked by neutralizing antibodies directed against Galphaq/11, coexpression of the regulatory domain of protein kinase Cdelta (PKCdelta), preactivating PKC with phorbol ester, or pharmacological suppression of PKC with bisindolylmaleimide I. Stimulation of CaV2.3 is Ca(2+)-independent and insensitive to 12-(2-cyanoethyl)-6,7,12,13 tetrahydro-13-methyl-5-oxo-5H-indolo(2,3-a)pyrrolo(3,4-c)-carbazole (Go 6976), a specific inhibitor of Ca(2+)-dependent PKC isozymes. These results indicate that muscarinic stimulation of CaV2.3 involves signaling by Galphaq/11, diacylglycerol, and a Ca(2+)-independent PKC. In contrast to stimulation, the magnitude of CaV2.3 inhibition depended on receptor subtype, with M3 and M5 receptors producing much larger CaV2.3 inhibition than M1 receptors. Interestingly, muscarinic inhibition of CaV2.3 was notably enhanced during pharmacological suppression of PKC, suggesting the presence of cross-talk between Gbetagamma-mediated inhibition and PKC-mediated stimulation of R-type channels similar to that described previously for N-type channels. PMID- 14742682 TI - Reverse transcriptase activity of hepatitis B virus (HBV) DNA polymerase within core capsid: interaction with deoxynucleoside triphosphates and anti-HBV L deoxynucleoside analog triphosphates. AB - The use of L(-)SddC [beta-L-2',3'-dideoxy-3'-thiacytidine (lamivudine, 3TC)] for the treatment of Herpes B virus (HBV) infection is hindered by the emergence of drug-resistance associated with the L526M, L550V, and L526M/M550V mutations of the viral DNA polymerase (DP). The interactions of the anti-HBV compounds 2',3' dideoxy-2',3'-didehydro-beta-L(-)-5-fluorode-oxycytidine and 2'-fluoro-5-methyl beta-L-arabinofuranosyluracil triphosphate with HBV DP and its L(-)SddC associated mutants have not been studied. The e antigen-negative variant of HBV associated with the G1896A mutation in the precore region has a high prevalence. Its effect on HBV DP is unclear. Because HBV DNA synthesis occurs in the nucleocapsid, we examined the kinetics of the reverse transcriptase activity from wild-type (wt) and mutated DPs with the wt or G1896A-mutated RNA template in the nucleocapsid. The effects of this template mutation on the activities of these L nucleoside triphosphates were also examined. Results indicated that these DP mutations increased the Km values of deoxy-NTPs and decreased the efficiencies (Vmax/Km) of DPs. The additional L526M mutation increased the efficiency of the M550V-mutated DP but no more than that of the L526M-mutated DP. The G1896A mutation had impacts on the interactions between different DPs and deoxy-NTPs, except dCTP. It also had different impacts on the actions of the L-nucleoside triphosphates toward DPs. The L526M and M550V mutations caused a greater decrease in the Vmax using the wt RNA template compared with the G1896A-mutated template. The L526M, M550V, and L526M/M550V mutations caused varying degrees of resistance to the different M-nucleoside triphosphates. PMID- 14742681 TI - Bucillamine induces the synthesis of vascular endothelial growth factor dose dependently in systemic sclerosis fibroblasts via nuclear factor-kappaB and simian virus 40 promoter factor 1 pathways. AB - The pathogenesis of systemic sclerosis (SSc) is characterized by activation of the immune system, impaired angiogenesis, and activated dermal fibroblasts. The effects of the immunosuppressive agent bucillamine (SA 96) on fibroblasts and angiogenic factors have not been examined. SA 96, and particularly its metabolite SA 981, increased the levels of vascular endothelial growth factor (VEGF) mRNA and protein dose-dependently in dermal fibroblasts from patients with SSc and healthy control subjects without influencing cell viability. SSc fibroblast cultures showed consistently a higher inducibility of VEGF than cultures from healthy control subjects. Preincubation with the SP-1 inhibitor mithramycin as well as blockade of nuclear factor (NF)-kappaB signaling with pyrrolidine dithiocarbamate treatment and IkappaB transfection reduced significantly the transcription of VEGF, indicating that both transcription factors contribute to the activation of VEGF by SA 981. Specific binding of NF-kappaB protein to its binding site after treatment with SA 981 was confirmed by electrophoretic mobility shift assay. In contrast, SA 981 did not influence the stability of VEGF mRNA as analyzed with actinomycin D assays. The study provides evidence for a role of NF-kappaB in the transcriptional regulation of the VEGF gene. SA 96 might have positive aspects on the impaired angiogenesis in patients with SSc. PMID- 14742683 TI - Vanadate is a potent activator of endothelial nitric-oxide synthase: evidence for the role of the serine/threonine kinase Akt and the 90-kDa heat shock protein. AB - We investigated the molecular mechanisms of sodium vanadate (vanadate)-induced nitric oxide (NO) production. Exposure of bovine lung microvascular cells (BLMVEC) to vanadate increased the release of biologically active NO in endothelium/smooth muscle cocultures, as measured by the accumulation of its surrogate marker, cGMP. This release was sensitive to NO synthase (NOS) inhibition and was greater than that observed with ionomycin. Although calcium chelators (BAPTA, EGTA) inhibited basal and ionomycin-induced NO production, they failed to inhibit vanadate-induced NO release. Moreover, in the absence of calcium/calmodulin, cell lysates from vanadate-treated cells exhibited greater NOS activity compared with control cells. Vanadate activates the phosphoinositide3-kinase (PI3-K)/Akt pathway, which is known to increase endothelial NOS (eNOS) activity by direct phosphorylation of Ser-1179. Treatment of BLMVEC with vanadate resulted in phosphorylation of both Akt and endothelial NOS. In addition, wortmannin, a PI3-K inhibitor, blocked both the vanadate induced phosphorylation of eNOS and the increase in cGMP accumulation. Similarly, adenovirus-mediated gene transfer of an activation deficient form of Akt (AA-Akt) blocked the release of NO brought about by vanadate. To further investigate the mechanism of action of vanadate, eNOS was immunoprecipitated and its association with proteins that alter eNOS activity was tested. Immunoblots demonstrated that the eNOS-caveolin interaction remained unaffected by vanadate, whereas vanadate promoted recruitment of the 90-kDa heat shock protein (hsp90) to eNOS. We conclude that vanadate causes NO release via a mechanism that involves Akt induced eNOS phosphorylation and increased binding of the activator protein hsp90 to eNOS. PMID- 14742684 TI - Different structural requirements of the ligand binding domain of the aryl hydrocarbon receptor for high- and low-affinity ligand binding and receptor activation. AB - The aryl hydrocarbon receptor (AhR) functions as a ligand-activated transcription factor that is responsible for the regulation of several response genes, of which the best characterized is the CYP1A1 gene. The present study was undertaken to elucidate the mechanism of activation of the AhR by omeprazole (OME), 2-mercapto 5-methoxybenzimidazole (MMB), and primaquine (PRQ), compounds that have previously been reported to induce CYP1A1 expression but that are not typical AhR ligands. All compounds caused a significant increase in luciferase activity in rat H4IIE and human HepG2 hepatoma cells transfected with a Gal4-AhR construct and the corresponding Gal4-Luc reporter gene. Furthermore, MMB and PRQ, but not OME, were capable of transforming cytosolic AhR to a DNA-binding form and displacing AhR-bound [3H]TCDD in rat hepatic cytosol in vitro. By performing site directed mutagenesis of residues in the ligand-binding domain of the Gal4-AhR, a construct containing a Y320F substitution was found to be resistant to activation by OME, MMB, and PRQ, but not by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Comparable affinities of [3H]TCDD-binding to the wild-type and the Y320F mutant Gal4-proteins, expressed in human embryonic kidney 293 cells, were obtained in the ligand-binding assay. In contrast, the competition of receptor-bound [3H]TCDD by PRQ was absent from Gal4-Y320F but not from Gal4-AhR cell extracts. The results of this study confirm that MMB and PRQ are low-affinity ligands for the AhR and suggest that high- and low-affinity ligands interact with different residues of the AhR ligand-binding pocket. In addition, the data presented here indicate that Tyr320 plays an important role in AhR activation. PMID- 14742685 TI - Agonist binding and Gq-stimulating activities of the purified human P2Y1 receptor. AB - The human P2Y1 receptor (P2Y1-R) was purified after high-level expression from a recombinant baculovirus in Sf9 insect cells. Quantification by protein staining and with a radioligand binding assay using the high-affinity P2Y1-R antagonist [3H]MRS2279 ([3H]2-chloro-N6-methyl-(N)-methanocarba-2'-deoxyadenosine 3',5'-bis phosphate) indicated a nearly homogenous preparation of receptor protein. Ki values determined in [3H]MRS2279 binding assays for antagonists with the purified P2Y1-R were in good agreement with the Ki and KB values determined for these molecules in membrane binding and activity assays, respectively. Availability of P2Y1-R in purified form allowed direct determination of nucleotide agonist affinities under conditions not compromised by nucleotide metabolism/interconversion, and an order of affinities of 2-methylthio-ADP (2MeSADP) > ADP = 2-methylthioATP = adenosine-5'-O-(3-thio)triphosphate = adenosine-5'-O(2-thiodiphosphate) >> ATP was obtained. The signaling activity of the purified P2Y1-R was quantified after reconstitution in proteoliposomes with heterotrimeric G proteins. Steady-state GTP hydrolysis in vesicles reconstituted with P2Y1-R and Galpha(q)beta(1)gamma(2) was stimulated by the addition of either 2MeADP or RGS4 alone and was increased by up to 50-fold in their combined presence. EC50 values of agonists for activation of the purified P2Y1-R were similar to their respective Ki values determined in radioligand binding experiments with the purified receptor. Moreover, ATP exhibited 20-fold higher EC50 and Ki values than did ADP and was a partial agonist relative to ADP and 2MeSADP under conditions in which no metabolism of the nucleotide occurred. Both RGS4 and PLC-beta1 were potent and efficacious GTPase-activating proteins for Galphaq and Galpha11 in P2Y1-R-containing vesicles. These results illustrate that the binding and signaling properties of the human P2Y1-R can be studied with purified proteins under conditions that circumvent the complications that occur in vivo. PMID- 14742686 TI - Systemic morphine administration suppresses genes involved in antigen presentation. AB - Administration of opioids in both humans and animal models results in significant alterations in immune system responsiveness. Although the majority of studies have focused on phenotypic changes in immune cells after short- and long-term morphine administration, few studies have determined whether alterations in gene expression profiles accompany these effects. To address this question, rats were treated with either morphine (20 mg/kg) or saline, and changes in gene expression and function in blood leukocytes were examined. Within 2 h, morphine administration resulted in a decrease in blood leukocyte expression of the major histocompatibility complex class II (MHC II RT1.B beta) (-3.27-fold) and related molecules, including the MHC II invariant chain (-2.73-fold). Furthermore, these changes in gene expression were accompanied by a significant decrease in surface MHC II RT1.B beta protein expression, specifically on B lymphocytes. Morphine administration was also found to inhibit IL-4 induced up-regulation of MHC II RT1.B beta cell surface expression on B lymphocytes. This is the first demonstration that receptors involved in antigen presentation are modified after systemic morphine administration. We propose that the inability of B lymphocytes to up-regulate key immune proteins, such as the MHC II molecule, after exposure to antigen-induced cytokine production may account for the increase in the susceptibility to bacterial and viral infections such as HIV in both drug abusers and patients receiving morphine. PMID- 14742687 TI - Two-pore-domain K+ channels are a novel target for the anesthetic gases xenon, nitrous oxide, and cyclopropane. AB - Nitrous oxide, xenon, and cyclopropane are anesthetic gases that have a distinct pharmacological profile. Whereas the molecular basis for their anesthetic actions remains unclear, they behave very differently to most other general anesthetics in that they have little or no effect on GABAA receptors, yet strongly inhibit the N-methyl-d-aspartate subtype of glutamate receptors. Here we show that certain members of the two-pore-domain K+ channel superfamily may represent an important new target for these gaseous anesthetics. TREK-1 is markedly activated by clinically relevant concentrations of nitrous oxide, xenon, and cyclopropane. In contrast, TASK-3, a member of this family that is very sensitive to volatile anesthetics, such as halothane, is insensitive to the anesthetic gases. We demonstrate that the C-terminal cytoplasmic domain is not an absolute requirement for the actions of the gases, although it clearly plays an important modulatory role. Finally, we show that Glu306, an amino acid that has previously been found to be important in the modulation of TREK-1 by arachidonic acid, membrane stretch and internal pH, is critical for the activating effects of the anesthetic gases. PMID- 14742688 TI - Pharmacological properties of alpha 9 alpha 10 nicotinic acetylcholine receptors revealed by heterologous expression of subunit chimeras. AB - The nicotinic acetylcholine receptor (nAChR) alpha9 and alpha10 subunits are expressed primarily within hair cells of the inner ear and have been implicated in auditory processing. Although functional recombinant nAChRs generated by the coexpression of alpha9 and alpha10 in Xenopus laevis oocytes have been described previously, there have been no reports of the successful heterologous expression of alpha9alpha10 nAChRs in cultured cell lines. In this study, subunit chimeras (alpha9chi and alpha10chi) have been constructed that contain the extracellular, ligand binding domain of the alpha9 or alpha10 subunits fused to the C-terminal domain of the 5-hydroxytryptamine type 3A (5HT3A) subunit. Specific high-affinity binding of the nicotinic radioligand [3H]methyllycaconitine was detected in membrane preparations of mammalian cells transfected with alpha9chi or alpha10chi alone, but significantly higher levels of binding were detected when alpha9chi and alpha10chi were cotransfected, providing evidence of a requirement for coassembly of alpha9 and alpha10 for the efficient formation of a nicotinic binding site. The pharmacological profile of alpha9chialpha10chi receptors, determined by equilibrium radioligand binding studies, is broadly similar to that determined previously by electrophysiological studies conducted with native and recombinant alpha9alpha10 nAChRs. In agreement with evidence that alpha9alpha10 nAChRs exhibit an atypical pharmacological profile, we have identified specific high-affinity binding of several non-nicotinic ligands including strychnine (a glycine receptor antagonist), bicuculline (a GABAA receptor antagonist), and atropine (a muscarinic acetylcholine receptor antagonist). Results have also been compared with radioligand binding data conducted with a previously described alpha7/5HT3A (alpha7chi) subunit chimera. PMID- 14742689 TI - Cytochrome P4501A1 promotes G1 phase cell cycle progression by controlling aryl hydrocarbon receptor activity. AB - The aryl hydrocarbon receptor (AhR) transcription factor is increasingly recognized as functioning in cell cycle control. Several recent reports have shown that AhR activity in the absence of exogenous agonists or presence of the prototypical ligand 2,3,7,8-tetrachlorodibenzo-p-dioxin can affect G1 phase progression in cultured cells. Serum release of serum-starved (G0) 5L rat hepatoma cells triggers transient AhR activation and P4501A1 protein expression concomitant with the G0/G1-to-S phase transition. In contrast, sustained AhR activation in response to TCDD treatment increases p27Kip1 expression in addition to P4501A1, resulting in G1 phase cell cycle arrest. Treating serum-released 5L cells with the alkyne metabolism-based P4501A1 inhibitor 1-(1-propynyl)pyrene results in prolonged AhR activation, enhanced p27Kip1 expression, and G1 phase arrest after serum release. The data are consistent with a cell cycle role for P4501A1 because they show that P4501A1 negatively regulates the duration of AhR action through the metabolic removal of the receptor agonist, thereby preventing AhR-mediated G1 phase arrest. PMID- 14742690 TI - Aspirin-mediated COX-2 transcript stabilization via sustained p38 activation in human intestinal myofibroblasts. AB - Acetylsalicylic acid (aspirin) is a cyclooxygenase (COX) inhibitor, yet some of its therapeutic effects are thought to derive from mechanisms unrelated to prostaglandin synthesis inhibition. In human intestinal myofibroblasts, aspirin, at therapeutic doses, had the unexpected effect of inducing prolonged COX-2 expression. This induction was especially pronounced when cells were treated with interleukin-1alpha (IL-1) plus aspirin for 24 h. Sodium salicylate, a poor COX inhibitor, likewise enhanced IL-1-mediated COX-2 gene expression whereas 5 aminosalicylic acid (5-ASA) or indomethacin had no effect. The COX-2 transcriptional rate, measured by nuclear runoff analysis and heterogeneous nuclear RNA reverse transcription-polymerase chain reaction, was only modestly elevated by aspirin treatment. In contrast, aspirin treatment dramatically stabilized the COX-2 message. The COX-2 mRNA half-life in IL-1 treated cells was 1 h and was increased in excess of 5 h in IL-1 + aspirin-treated cells. Phosphorylation of p38 MAPK was enhanced in aspirin-treated cells (but not in cells treated with 5-ASA or indomethacin) for up to 24 h after treatment. Inhibition of p38 activity negated aspirin-mediated COX-2 mRNA stabilization and the resultant increase in COX-2 mRNA and protein levels. The modest transcriptional response seen in aspirin treated cells was also abolished by p38 inhibition. We conclude that aspirin enhances COX-2 expression via sustained activation of p38, which results in prolonged stabilization of the COX-2 message and a slightly elevated transcription rate. Aspirin also enhanced steady-state mRNA levels of other IL-1 modulated genes (IL-1beta, IL-6, groalpha, and TNFalpha) that are likewise regulated at the level of message stability via p38 activation. PMID- 14742691 TI - Anti-Mullerian hormone expression pattern in the human ovary: potential implications for initial and cyclic follicle recruitment. AB - Anti-Mullerian hormone (AMH) is a member of the transforming growth factor-beta superfamily, which plays an important role in both ovarian primordial follicle recruitment and dominant follicle selection in mice. However, the role of AMH in folliculogenesis in humans has not been investigated in detail. In the present study, AMH expression was assessed using immunohistochemistry in ovarian sections, obtained from healthy regularly cycling women. To this end, a novel monoclonal antibody to human AMH was developed. AMH expression was not observed in primordial follicles, whereas 74% of the primary follicles showed at least a weak signal in the granulosa cells. The highest level of AMH expression was present in the granulosa cells of secondary, preantral and small antral follicles 0.20). We suggest therefore that any potential role of the carotid bodies in glucose homeostasis in vivo is mediated through its transduction of some other metabolically derived blood-borne factor rather than glucose per se and that this may also provide the link between exercise, metabolic rate and ventilation. PMID- 14742729 TI - Congenital hypoventilation and impaired hypoxic response in Nurr1 mutant mice. AB - Nurr1, a transcription factor belonging to the family of nuclear receptors, is expressed at high levels immediately after birth. Gene-targeted mice lacking Nurr1 fail to develop midbrain dopaminergic neurones and do not survive beyond 24 h after birth. Dopamine (DA) levels may be regulated by Nurr1, and as DA is involved in both central and peripheral respiratory control, we hypothesized that lack of Nurr1 may impair breathing and cause death by respiratory failure. We demonstrate herein that Nurr1 newborn knockout mice have a severely disturbed breathing pattern characterized by hypoventilation, numerous apnoeas and failure to increase breathing when challenged with hypoxia. In heterozygote Nurr1 mice the response to hypoxia is also altered. Furthermore, the central respiratory rhythm, generated from isolated brainstem-spinal cord preparations, exhibits impaired response to hypoxia in mice lacking Nurr1. Moreover, Nurr1 is expressed in several respiratory-related regions of the nervous system, including the nucleus of the solitary tract, the nucleus ambiguus and the dorsal motor nucleus of the vagus nerve, and in the carotid bodies. The prominent Nurr1 expression in these areas, involved in respiratory control, along with the severe respiratory phenotype, indicates that Nurr1 plays a major role in the extrauterine adaption of respiratory control and the response to hypoxia. PMID- 14742730 TI - Functional up-regulation of KCNA gene family expression in murine mesenteric resistance artery smooth muscle. AB - This study focused on the hypothesis that KCNA genes (which encode K(V)alpha1 voltage-gated K(+) channels) have enhanced functional expression in smooth muscle cells of a primary determinant of peripheral resistance - the small mesenteric artery. Real-time PCR methodology was developed to measure cell type-specific in situ gene expression. Profiles were determined for arterial myocyte expression of RNA species encoding K(V)alpha1 subunits as well as K(V)beta1, K(V)alpha2.1, K(V)gamma9.3, BK(Ca)alpha1 and BK(Ca)beta1. The seven major KCNA genes were expressed and more readily detected in endothelium-denuded mesenteric resistance artery compared with thoracic aorta; quantification revealed dramatic differential expression of one to two orders of magnitude. There was also four times more RNA encoding K(V)alpha2.1 but less or similar amounts encoding K(V)beta1, K(V)gamma9.3, BK(Ca)alpha1 and BK(Cabeta)1. Patch-clamp recordings from freshly isolated smooth muscle cells revealed dominant K(V)alpha1 K(+) current and current density twice as large in mesenteric cells. Therefore, we suggest the increased RNA production of the resistance artery impacts on physiological function, although there is quantitatively less K(+) current than might be expected. The mechanism conferring up-regulated expression of KCNA genes may be common to all the gene family and play a functional role in the physiological control of blood pressure. PMID- 14742731 TI - Single-channel recordings of a rapid delayed rectifier current in adult mouse ventricular myocytes: basic properties and effects of divalent cations. AB - The rapidly delayed rectifier current (I(Kr)) has been described in ventricular myocytes isolated from many species, as well as from neonatal mice. However, whether I(Kr) is present in the adult mouse heart remains controversial. We used cell-attached patch-clamp recording in symmetrical K(+) solutions to assess the presence and behaviour of single I(Kr) channels in adult mouse cardiomyocytes (mI(Kr)). Of 314 patches, 158 (50.1%) demonstrated mI(Kr) currents as compared with 131 (42.3%) for the I(K1) channel. Single mI(Kr) channel activity was rarely observed at potentials positive to -10 mV. The slope conductance at negative potentials was 12 pS. Upon repolarization, ensemble-averaged mI(Kr) showed slow deactivation with a biexponential time course. A selective I(Kr) blocker, E-4031 (1 microm), completely blocked mI(Kr) channel activity. Extracellular Ca(2+) and Mg(2+) at physiological concentrations shifted the activation by approximately 30 mV, accelerated deactivation kinetics, prolonged long-closed time, and reduced open probability without affecting single-channel conductance, suggesting a direct channel-blocking effect in addition to well-recognized voltage shifts. HERG subunits expressed in Chinese hamster ovary cells produced channels with properties similar to those of mI(Kr), except for the more-negative activation of the HERG channels. Despite the abundant expression of mI(Kr), single-channel events were rarely observed during action-potential clamp and 5 microm E-4031 had no detectable effect on the action potential parameters, confirming that mI(Kr) plays at best a minor role in repolarization of adult mouse cardiomyocytes, probably because the modulatory effects of divalent cations prevent significant mI(Kr) opening under physiological conditions. PMID- 14742732 TI - Protein phosphatase 1 and an opposing protein kinase regulate steady-state L-type Ca2+ current in mouse cardiac myocytes. AB - Studies have suggested that integration of kinase and phosphatase activities maintains the steady-state L-type Ca(2+) current in ventricular myocytes, a balance disrupted in failing hearts. As we have recently reported that the PP1/PP2A inhibitor calyculin A evokes pronounced increases in L-type I(Ca), the goal of this study was to identify the counteracting kinase and phosphatase that determine 'basal'I(Ca) in isolated mouse ventricular myocytes. Whole-cell voltage clamp studies, with filling solutions containing 10 mm EGTA, revealed that calyculin A (100 nm) increased I(Ca) at test potentials between -42 and +49 mV (44% at 0 mV) from a holding potential of -80 mV. It also shifted the V(0.5) (membrane potential at half-maximal) of both activation (from -17 to -25 mV) and steady-state inactivation (from -32 to -37 mV) in the hyperpolarizing direction. The broad-spectrum protein kinase inhibitor, staurosporine (300 nm), was without effect on I(Ca) when added after calyculin A. However, by itself, staurosporine decreased I(Ca) throughout the voltage range examined (50% at 0 mV) and blocked the response to calyculin A, indicating that the phosphatase inhibitor's effects depend upon an opposing kinase activity. The PKA inhibitors Rp-cAMPs (100 microm in the pipette) and H89 (1 microm) failed to reduce basal I(Ca) or to block the calyculin A-evoked increase in I(Ca). Likewise, calyculin A was still active with 10 mm intracellular BAPTA or when Ba(2+) was used as the charge carrier. These data eliminate roles for protein kinase A (PKA) and calmodulin-dependent protein kinase II (CaMKII) as counteracting kinases. However, the protein kinase C (PKC) inhibitors Ro 31-8220 (1 microm) and Go 6976 (200 nm) decreased steady-state I(Ca) and blunted the effect of calyculin A. PP2A is not involved in this regulation as intracellular applications of 10-100 nm okadaic acid or 500 nm fostriecin failed to increase I(Ca). However, PP1 is important, as dialysis with 2 microm okadaic acid or 500 nm inhibitor-2 mimicked the increases in I(Ca) seen with calyculin A. These in situ studies identify constitutive activity of PP1 and the counteracting activity of certain isoforms of PKC, in pathways distinct from receptor-mediated signalling cascades, as regulatory components that determine the steady-state level of cardiac L-type I(Ca). PMID- 14742733 TI - The effect of tropomyosin on force and elementary steps of the cross-bridge cycle in reconstituted bovine myocardium. AB - The role of tropomyosin (Tm) in the elementary steps of the cross-bridge cycle in bovine myocardium was investigated. The thin filament was selectively removed using gelsolin (thin filament severing protein), and the actin filament was reconstituted from G-actin. Tm was further reconstituted without troponin (Tn), and the kinetic constants of the elementary steps of the cross-bridge cycle were deduced using sinusoidal analysis at pCa 2-methylthioadenosine-5' triphosphate > ADP with an EC(50) of 30 nM, 0.2 microM, and 0.8 microM, respectively. Activation of P2Y1R increased [Ca(2+)](i), which was prevented by the P2Y1R antagonists pyridoxalphosphate-6-azophenyl-2',4'-disulfonic acid (PPADS) (10 microM) and N6-methyl 2'-deoxyadenosine 3',5'-bisphosphate (MRS2179) (10 microM) and by pretreatment with P2Y1R antisense oligodeoxynucleotides. In CHO-K1 and CHO-KNUT (mock-transfected) cells lacking CFTR, both P2Y1R and P2Y2R caused [Ca(2+)](i) mobilization via pertussis toxin (PTX)-insensitive G(q/11) proteins. In contrast, in CFTR-expressing CHO cells (CHO-BQ1), the P2Y1R response was completely PTX-sensitive, indicating that P2Y1R couples to G(i/o)-proteins, whereas the P2Y2R response remained PTX-insensitive. In CHO-BQ1 cells, P2Y1R activation by ADP (100 microM) failed to inhibit both forskolin (1 microM) induced CFTR activation, measured using iodide ((125)I) efflux, and forskolin (0.1-10 microM)-evoked cAMP increase. Together, our results indicate that, in contrast to P2Y2R, P2Y1R does not modulate CFTR activity in CHO cells and that CFTR expression may alter the G-protein-coupling selectivity of P2Y1R. PMID- 14742737 TI - Bremazocine increases C-type natriuretic peptide levels in aqueous humor and enhances outflow facility. AB - A relatively selective agonist of kappa opioid receptors (KOR), bremazocine (BRE), lowers intraocular pressure in rabbits, in part, by increasing natriuretic peptide levels in aqueous humor and by enhancing total outflow facility (TOF). Natriuretic peptide (NP) levels [atrial NP (ANP), brain NP (BNP), and C-type NP (CNP)] were measured in aqueous humor of rabbits either by radioimmunoassay or enzyme immunoassay. TOF was determined in rabbits by two-level constant pressure perfusion of the anterior chamber. Experimental regimens included topical treatment with BRE in the presence or absence of KOR antagonist (norbinaltorphimine), protein kinase C inhibitor (chelerythrine), and natriuretic peptide receptor antagonist (isatin). The rank order of basal NP levels in aqueous humor of rabbits was BNP CNP > ANP. Topical administration of BRE (1-100 microg) caused dose-related elevations of CNP levels in aqueous humor that were inhibited by topical pretreatment with either norbinaltorphimine (100 microg, bilaterally) or chelerythrine (10 microg, bilaterally). Topically administered BRE (100 microg) also elevated levels of ANP and BNP in aqueous humor and evoked an 80% increase in TOF. The increase in TOF was antagonized by topical pretreatment with either norbinaltorphimine (100 microg, bilaterally) or isatin (100 microg, bilaterally). Bremazocine induced an increase in NP (ANP, BNP, and CNP) levels and TOF in rabbits by activating KOR. The increase in CNP levels elicited by BRE was inhibited by norbinaltorphimine and chelerythrine; therefore, this event is most likely mediated by a KOR-linked activation of protein kinase C. These data provide evidence that the increase in TOF elicited by BRE was mediated by a KOR-activated paracrine effect of NPs on tissues within ocular outflow tract(s). PMID- 14742738 TI - Enantioselectivity of alpha-benzyl-alpha-methyl-gamma-butyrolactone-mediated modulation of anticonvulsant activity and GABA(A) receptor function. AB - Alkyl-substituted butyrolactones have both inhibitory and stimulatory effects on GABA(A) receptors. Lactones with small alkyl substitutions at the alpha-position positively modulate the channel, whereas beta-substituted lactones tend to inhibit the GABA(A) receptor. These compounds mediate inhibition through the picrotoxin site of the receptor. A distinct binding site that mediates the stimulatory actions of lactones is presumed to exist, although no definitive evidence to support this claim exists. In the present study, we used in vivo and in vitro assays to evaluate the effects of the enantiomers of a novel lactone, alpha-benzyl-alpha-methyl-gamma-butyrolactone (alpha-BnMeGBL), on the GABA(A) receptor. R-(-)-alpha-BnMeGBL was 2-fold more potent than the S-(+)-alpha-BnMeGBL in blocking pentylenetetrazol-induced seizures in CF-1 mice. The (+)-enantiomer inhibited binding of t-butylbicyclophosporothionate with a higher affinity than the (-)-enantiomer (IC(50) of 0.68 and 1.1 mM, respectively). Whole cell patch clamp recordings from recombinant alpha1beta2gamma2 receptors stably expressed in HEK293 cells demonstrated that both compounds stimulated GABA-activated current. The maximal stimulation was approximately 2-fold greater with (+)-alpha-BnMeGBL than that seen with (-)-alpha-BnMeGBL. Both enantiomers of alpha-BnMeGBL directly gated the GABA(A) receptor at mM concentrations, in a nonstereoselective manner. Our data demonstrate the stimulatory actions of alpha-BnMeGBL on GABA(A) receptor function display enantioselectivity and provide strong evidence for the existence of a true "lactone site" on the receptor. PMID- 14742739 TI - The Discriminative stimulus effects of gamma-hydroxybutyrate and related compounds in rats discriminating baclofen or diazepam: The role of GABA(B) and GABA(A) receptors. AB - The discriminative stimulus effects of gamma-hydroxybutyrate (GHB) can be mimicked by GABA(A) receptor-positive modulators (e.g., diazepam) and GABA(B) receptor agonists (e.g., baclofen). The purposes of this study were to see whether stimulus control could be established with baclofen and to further characterize the role of GABAergic mechanisms in the behavioral actions of GHB by evaluating GHB and related compounds in rats discriminating either diazepam or baclofen. Training criteria were satisfied with baclofen and diazepam after 69 and 44 sessions, respectively. GHB and its precursors gamma-butyrolactone and 1,4 butanediol occasioned >80% responding on the drug-associated lever in rats discriminating baclofen and <11% in rats discriminating diazepam. Diazepam and other GABA(A) receptor-positive modulators occasioned intermediate levels of responding on the baclofen lever, whereas baclofen occasioned less than 4% responding on the diazepam lever. The GABA(B) receptor antagonist CGP 35348 [(3 aminopropyl)(diethoxymethyl) phosphinic acid] partially antagonized the effects of baclofen as well as the baclofen-like effects of GHB, and flumazenil partially antagonized the effects of diazepam. This study established stimulus control with baclofen, and substitution data provided direct evidence for a role of GABAergic, especially GABA(B), mechanisms in the discriminative stimulus effects of GHB. The lack of substitution by GHB or its metabolic precursors for diazepam indicates a comparatively smaller role of GABA(A) mechanisms in these effects of GHB. The inability of CGP 35348 to completely attenuate the effects of baclofen and GHB suggests that multiple receptors could be involved in the discriminative stimulus effects of GHB. PMID- 14742740 TI - Overexpression of pituitary adenylate cyclase-activating polypeptide in islets inhibits hyperinsulinemia and islet hyperplasia in agouti yellow mice. AB - Pituitary adenylate cyclase-activating polypeptide (PACAP) is an intraislet neuropeptide and shares insulinotropic and insulin-sensitizing properties with glucagon-like peptide-1 (GLP-1); however, the pathophysiological significance of PACAP in diabetes remains largely unknown. To assess this, we crossed our recently developed transgenic mice overexpressing PACAP in pancreatic beta-cells (Tg/+), with lethal yellow agouti (KKA(y)) mice (A(y)/+), a genetic model for obesity-diabetes, and examined the metabolic and morphological phenotypes of F(1) animals. Tg/+ mice with the A(y) allele (Tg/+:A(y)/+) developed maturity-onset obesity and diabetes associated with hyperglycemia, hyperlipidemia, and hyperphagia, similar to those of A(y)/+ mice, but hyperinsulinemia was significantly ameliorated in Tg/+:A(y)/+ mice. Although A(y)/+ mice exhibited a marked increase in islet mass resulting from hyperplasia and hypertrophy, this increase was significantly attenuated in Tg/+:A(y)/+ mice. Size frequency distribution analysis revealed that the very large islets comprising one-fourth of islets of A(y)/+ mice were selectively reduced in Tg/+:A(y)/+ mice. Because functional defects have been demonstrated in the large islets of obese animal models, together these findings suggest that PACAP regulates hyperinsulinemia and the abnormal increase in islet mass that occurs during the diabetic process. PMID- 14742741 TI - Novel ceramide analogs as potential chemotherapeutic agents in breast cancer. AB - Recent evidence suggests a role for aberrant ceramide levels in the pathogenesis of cancer and chemoresistance and indicates that manipulation of tumor ceramide levels may be a useful strategy in the fight against breast cancer. This study demonstrates that alterations in the degree and position of unsaturation of bonds in the sphingoid backbone of d-erythro-N-octanoyl-sphingosine (Cer) affect the antiproliferative ability of ceramide analogs in breast cancer cells. The most potent analog of Cer we tested is (2S,3R)-(4E,6E)-2-octanoylamidooctadecadiene 1,3-diol (4,6-diene-Cer), which contains an additional trans double bond at C(6) C(7) of the sphingoid backbone. 4,6-Diene-Cer exhibited higher potency than Cer in tumor necrosis factor (TNF)-alpha-resistant (IC(50) of 11.3 versus 32.9 microM) and TNF-alpha-sensitive (IC(50) of 13.7 versus 37.7 microM) MCF-7 cells. 4,6-Diene-Cer was also more potent than Cer in inducing cell death in MDA-MB-231 and NCI/ADR-RES breast cancer cell lines (IC(50) of 3.7 versus 11.3 microM, and 24.1 versus 86.9 microM, respectively). 4,6-Diene-Cer caused a prolonged elevation of intracellular ceramide levels in MCF-7 cells, which may contribute to its enhanced cytotoxicity. Furthermore, treatment of MCF-7 cells with Cer or 4,6-diene-Cer resulted in induction of apoptosis by 8 h via the mitochondrial pathway, as demonstrated by release of cytochrome c, loss of membrane asymmetry (measured by Annexin V staining), and a decrease in the mitochondrial membrane potential. Importantly, both Cer and 4,6-diene-Cer displayed selectivity toward transformed breast cells over nontransformed breast epithelial cells. These data suggest that these and other novel ceramide analogs represent potential therapeutic agents in breast cancer treatment. PMID- 14742742 TI - Characterization of IDN-6556 (3-[2-(2-tert-butyl-phenylaminooxalyl)-amino] propionylamino]-4-oxo-5-(2,3,5,6-tetrafluoro-phenoxy)-pentanoic acid): a liver targeted caspase inhibitor. AB - The potency, efficacy, and pharmacokinetic properties of IDN-6556 (3-[2-[(2-tert butyl-phenylaminooxalyl)-amino]-propionylamino]-4-oxo-5-(2,3,5,6-tetrafluoro phenoxy)-pentanoic acid), a first-in-class caspase inhibitor in clinical trials for the treatment of liver diseases, were characterized in vivo in rodent models. In the mouse alpha-Fas model of liver injury, i.p. administration of IDN-6556 resulted in marked reduction of alanine aminotransferase (ALT), apoptosis, and caspase activities at a dose of 3 mg/kg. At this dose, IDN-6556 was also effective when given up to 2 h before alpha-Fas and as late as 4 h after alpha Fas administration. In both the alpha-Fas and d-galactosamine/lipopolysaccharide (D-Gln/LPS) model, ED(50) values in the sub-milligram per kilogram range were established after a number of routes of administration (i.p., i.v., i.m., or p.o.), ranging from 0.04 to 0.38 mg/kg. Efficacy was also demonstrated in the rat D-Gln/LPS model with 67 and 72% reductions in ALT activities after i.p. and p.o. treatment with IDN-6556 (10 mg/kg), respectively. Pharmacokinetic analysis in the rat demonstrated rapid clearance after i.v., i.p., and s.c. administration with terminal t(1/2) ranging from 46 to 51 min. Low absolute bioavailability after p.o. administration was seen (2.7-4%), but portal drug concentrations after oral administration were 3-fold higher than systemic concentrations with a 3.7-fold increase in the terminal t(1/2), indicating a significant first-pass effect. Liver concentrations remained constant after oral administration for at least a 4 h period, reaching a C(max) of 2558 ng/g liver at 120 min. Last, 51 +/- 20 and 4.9 +/- 3.4% of IDN-6556 was excreted intact in bile after i.v. and p.o. administration, respectively. This evaluation indicates that IDN-6556 has marked efficacy in models of liver disease after oral administration and thus, is an excellent candidate for the treatment of liver diseases characterized by excessive apoptosis. PMID- 14742743 TI - Role of adenosine A2A receptor in the regulation of gastric somatostatin release. AB - Adenosine has been demonstrated to inhibit gastric acid secretion. In the rat stomach, this inhibitory effect may be mediated indirectly by increasing the release of somatostatin-like immunoreactivity (SLI). Results show that adenosine analogs augmented SLI release in the isolated vascularly perfused rat stomach. The rank order of potency of the analogs in stimulating SLI release was 2-p-(2 carboxyethyl)phenethylamino-5'-N-ethylcarboxamidoadenosine (CGS 21680) approximately 5'-N-ethylcarboxamidoadenosine > 2-chloroadenosine > R-(-)-N(6)-(2 phenylisopropyl)adenosine >1-deoxy-1-[6-[[(3-iodophenyl)methyl]amino]-9H-purin-9 yl]-N-methyl-beta-d-ribofuranuronamide > N(6)-cyclopentyladenosine approximately N(6)-cyclohexyladenosine > S-(+)-N(6)-(2-phenylisopropyl) adenosine, suggesting the involvement of the A(2A) receptor. In agreement, 4-(2-[7-amino-2-(2 furyl)[1,2,4]triazolo[2,3-a] [1,3,5]triazin-5-ylamino]ethyl)phenol (ZM 241385), an A(2A) receptor antagonist, was shown to abolish the adenosine- and CGS 21680 stimulated SLI release. Immunohistochemical studies reveal the presence of A(2A) receptor immunoreactivity on the gastric plexi and mucosal D-cells, but not on parietal cells and G-cells, suggesting that adenosine may act directly on D-cells or indirectly on the gastric plexi to augment SLI release. The present study also demonstrates that the structure of the mucosal A(2A) receptor is identical to that in the rat brain, and that alternative splicing of this gene does not occur. A real-time reverse transcription-polymerase chain reaction assay has also been established to quantify the levels of A(2A) receptor mRNA. Results show that gastric tissues contained significantly lower levels of A(2A) receptor mRNA compared with the striatum. The lowest level was detected in the mucosa. In conclusion, adenosine may act on A(2A) receptors to augment SLI release and consequently control gastric acid secretion. PMID- 14742744 TI - Extracellular signal-regulated kinase/mitogen-activated protein kinases block internalization of delta-opioid receptors. AB - Translocation of G protein-coupled receptors (GPCRs) from the cell membrane to cytosol depends on the kind of ligand activating the receptor. This principle is clearly demonstrated for opioid receptors, because diverse opiate agonists rapidly induce receptor internalization, whereas morphine almost fails. We report here the impact of mitogen-activated protein (MAP) kinase isoforms extracellular signal-regulated kinase (ERK)1/2 on the internalization of delta-opioid receptors (DORs) expressed in human embryonic kidney (HEK)293 cells. Receptor activation by etorphine turned out to transiently phosphorylate ERK/MAP kinases and bring about DOR internalization within 20 min. In contrast, prolonged exposure of HEK293 cells to morphine excited persistent phosphorylation of ERK/MAP kinases, and those cells failed to internalize the opioid receptor. When ERK/MAP kinase phosphorylation was blocked by 2'-Amino-3'-methoxyflavone (PD98059), morphine gained the ability to strongly induce DOR endocytosis. The importance of activated MAP kinases for DOR internalization is further demonstrated by glutamate and paclitaxel because these substances induce phosphorylation of ERK1/2 and concomitantly prevent DOR sequestration by etorphine. In addition, receptor internalization by morphine was facilitated by inhibition of protein kinase C and opioid-mediated transactivation of epidermal growth factor receptor (EGFR), both activating ERK/MAP kinases by opioids. The mechanism affording DOR internalization by PD98059 may relate to arrestin, which uncouples GPCRs and thus triggers receptor internalization. Arrestin considerably translocates toward the cell membrane upon DOR activation by morphine in presence of the MAP kinase blocker, but it fails in the absence of PD98059. We conclude that ERK/MAP kinase activity prevents opioid receptor desensitization and sequestration by blocking arrestin 2 interaction with activated DORs. PMID- 14742745 TI - Hammerhead ribozyme-mediated sensitization of human tumor cells after treatment with 1,3-bis(2-chloroethyl)-1-nitrosourea. AB - O(6)-Methylguanine DNA methyltransferase (MGMT) protects tumor cells from the cytotoxic effects of DNA-alkylating agents such as 1,3-bis-(2-chloroethyl)-1 nitrosourea (BCNU). To improve the therapeutic index of BCNU, biochemical strategies to inhibit MGMT temporarily by systemic administration of small molecules, such as O(6)-benzylguanine, have been developed and are showing promise in clinical trials. In this study, an alternative molecular strategy for modulating BCNU resistance was explored using hammerhead ribozymes (Rz) designed to degrade the long-lived MGMT mRNA. We had previously identified several ribozymes capable of decreasing MGMT levels in HeLa cells. Using colony formation assays, the BCNU-induced cell kill was shown to be increased by 1 to 3 logs in the HeLa/Rz clones compared with wild-type HeLa cells at a BCNU dose of 100 microM. In the current study, 10 randomly selected clones of Rz161, 212, and a reconstructed Rz178/212 were assayed for MGMT activity, MGMT mRNA, and sensitivity to BCNU. The 30 clones exhibited almost identical results in the three assays, i.e., nearly undetectable MGMT activity, greatly diminished MGMT mRNA, and comparable sensitivity to BCNU using the 4-[3-(4-iodophenyl)-2-(4 nitrophenyl)-2H-5-tetrazolio]-1,3-benzene disulfonate (WST-1) viability assay. The effects of catalytically inactive ribozymes carrying a single point mutation were compared with their active counterparts in vitro and in stably transfected clones to determine whether antisense inhibition was a contributor to the inhibition of MGMT activity we observed. Collectively, these results suggest that the hammerhead ribozymes characterized in this study will be excellent candidates for future gene therapy approaches targeting MGMT. PMID- 14742746 TI - Age-related differences in CYP3A expression and activity in the rat liver, intestine, and kidney. AB - We evaluated the effect of age on CYP3A expression and function in the liver, intestine, and kidney from young (3-4 months), intermediate (13-14 months), and old (25-26 months) male Fischer-344 rats. The biotransformation of triazolam to its primary hydroxylated products, 4-OH-TRZ (triazolam) and alpha-OH-TRZ, was used as a marker of CYP3A activity in rat liver and intestine. Immunoactive CYP3A expression was evaluated by Western blot analysis in the rat intestine, liver, and kidney. Since testosterone and NADPH reductase levels may modulate CYP3A activity, we also examined free plasma testosterone concentrations and NADPH reductase expression in these rats. The effect of age on CYP3A expression was tissue-specific. Although both CYP3A activity and expression were reduced by approximately 50 to 70% in the old livers compared with the young animals, intestinal CYP3A activity and expression did not change significantly with age. The expression of one CYP3A isoform was increased by 1.5-fold in the old kidneys. NADPH reductase expression was reduced by 23 to 36% with age in all tissues; this reached statistical significance only in the liver. Plasma testosterone levels declined by 74% in the old animals. This study suggests that the effect of age on CYP3A expression and function is tissue-specific. In addition, changes in testosterone levels and NADPH reductase expression may contribute to age-related differences in hepatic CYP3A activity. PMID- 14742747 TI - Evidence for multiple P2Y receptors in trabecular meshwork cells. AB - The purpose of this study was to determine whether functional purinergic P2 receptors are present in trabecular meshwork cells. The human trabecular cell line HTM-3 and cultured bovine trabecular cells were used to assess the effects of P2 agonists on intracellular Ca(2+) levels, extracellular signal-regulated kinase (ERK1/2) activation, and P2Y receptor expression. ATP, UTP, ADP, and 2 methyl-thio-adenosine triphosphate (2-MeS-ATP) each produced a concentration dependent increase in intracellular Ca(2+) in bovine trabecular cells and the HTM 3 cell line. The addition of UDP did not produce any detectable rise in intracellular Ca(2+). Pretreatment with the P2Y(1) receptor antagonist 2'-deoxy N(6)-methyladenosine-3',5'-diphosphate (MRS-2179) blocked the ADP- and 2-MeS-ATP induced rise in intracellular Ca(2+). However, the ATP- or UTP-induced rise in intracellular Ca(2+) was not inhibited by MRS-2179 pretreatment. The addition of ADP, 2-MeS-ATP, ATP, or UTP were also found to activate the ERK1/2 signaling pathway. This activation of ERK1/2 was blocked by pretreatment with the mitogen activated protein kinase kinase inhibitor 1,4-diamino-2,3-dicyano-1,4-bis(o aminophenylmercapto)butadiene (U-0126) or the protein kinase C inhibitor chelerythrine chloride, but not by MRS-2179. Analysis of mRNA from HTM-3 cells by reverse transcription-polymerase chain reaction revealed the expression of P2Y(1), P2Y(4), and P2Y(11) receptor subtypes. These data demonstrate that multiple P2Y receptors are present in trabecular cells. Our results are consistent with the idea that the mobilization of intracellular Ca(2+)results from the activation of P2Y(1) and P2Y(4) receptors, whereas the activation of the ERK1/2 pathway results from the activation of P2Y(4) receptors alone. However, a role for the P2Y(11) receptors in mobilization of Ca(2+), or activation of the ERK1/2 pathway, cannot be discounted. PMID- 14742748 TI - Pharmacokinetics and tissue distribution of the stereoisomers of 4-methylaminorex in the rat. AB - 4-Methylaminorex, a potential psychostimulant drug of abuse, exists as four stereoisomers: cis-4R,5S, cis-4S,5R, trans-4S,5S, and trans-4R,5R, which were shown previously to possess stereospecific effects. This study characterized their pharmacokinetic and tissue distribution profiles, and metabolic turnover to norephedrine and norpseudoephedrine, in male Wistar rats. The rats received each isomer intravenously, intraperitoneally, or orally, followed by blood sample collection via cannula (pharmacokinetic study), or tissue sample collection at predetermined time points (tissue distribution study). The samples were analyzed for cis- and trans-isomers, and when appropriate for norephedrine and norpseudoephedrine, with gas chromatography/mass spectrometry. Trans-4S,5S-, cis 4R,5S-, and cis-4S,5R-isomers behaved comparably kinetically (volume of distribution 1.7-2.3 l/kg, distribution half-life 3.8-7.0 min, elimination half life 35-42 min, and bioavailability 32-57% intraperitoneally or 4-16% orally), whereas trans-4R,5R-isomer differed from the others, with a longer elimination half-life (118-169 min) and higher bioavailability (100% intraperitoneally or 83% orally). The highest isomer concentrations were observed in the kidney followed most frequently by the liver, brain, muscle, and last by fat and blood. The elimination half-lives of the stereoisomers from the tissues were generally similar to those in blood. No pharmacologically significant amounts of norephedrine or norpseudoephedrine were detected in blood or the brain. In conclusion, differences between the stereoisomers of 4-methylaminorex in the pharmacokinetics and tissue distribution are described. However, these differences are not compatible with, and thus may not account for, the distinct behavioral and neurochemical effects of the stereoisomers demonstrated previously. Furthermore, metabolic turnover to norephedrine and norpseudoephedrine does not seem to contribute significantly to 4-methylaminorex pharmacology. PMID- 14742749 TI - Comparison of the effects of dextromethorphan, dextrorphan, and levorphanol on the hypothalamo-pituitary-adrenal axis. AB - Dextromethorphan is a weak noncompetitive N-methyl-D-aspartate (NMDA) receptor antagonist. It is metabolized in vivo to dextrorphan, a more potent noncompetitive NMDA antagonist that is the dextrorotatory enantiomer of the opioid agonist levorphanol. The present study characterized the effects of the acute administration of dextromethorphan, dextrorphan, and levorphanol on the hypothalamo-pituitary-adrenal (HPA) axis in the rat and tested the involvement of opioid receptors in the responses produced by dextrorphan and levorphanol. Although both dextromethorphan and dextrorphan increased plasma levels of adrenocorticotropic hormone (ACTH) and corticosterone, the dextromethorphan induced responses occurred more rapidly than the dextrorphan-induced responses. The analysis of plasma levels of dextrorphan produced after the administration of dextromethorphan indicates that the concentration of dextrorphan formed was too low to be pharmacologically relevant, suggesting that at least some of the effects on the HPA axis are due to the parent compound, and not the metabolite. Naloxone (2 mg/kg) had no effect on the dextrorphan-induced increases in plasma levels of ACTH and corticosterone, but it blocked the levorphanol-induced increases. These results support the hypothesis that dextromethorphan has pharmacological activity aside from its biotransformation to dextrorphan and demonstrate that the effects of dextrorphan are not mediated by opioid receptors. PMID- 14742750 TI - Anxiolytic-like effects of the corticotropin-releasing factor1 (CRF1) antagonist DMP904 [4-(3-pentylamino)-2,7-dimethyl-8-(2-methyl-4-methoxyphenyl)-pyrazolo-[1,5 a]-pyrimidine] administered acutely or chronically at doses occupying central CRF1 receptors in rats. AB - Corticotropin-releasing factor(1) (CRF(1)) antagonists may be effective in the treatment of anxiety disorders with fewer side effects compared with classic benzodiazepines. The behavioral effects of DMP904 [4-(3-pentylamino)-2,7-dimethyl 8-(2-methyl-4-methoxyphenyl)-pyrazolo-[1,5-a]-pyrimidine] and its effects on the hypothalamic-pituitary-adrenal (HPA) axis were related to its levels in plasma and estimated occupancy of central CRF(1) receptors. DMP904 (10-30 mg/kg, p.o.) and alprazolam (10 mg/kg, p.o.) increased time spent in open arms of an elevated plus maze. In addition, acutely or chronically (14 days) administered DMP904 (1.0 30 mg/kg, p.o.) and acute alprazolam (1.0-3.0 mg/kg, p.o.) significantly reduced exit latency in the defensive withdrawal model of anxiety in rats, suggesting that tolerance may not develop to the anxiolytic-like effects of DMP904 in this model of anxiety. Acutely, DMP904 reversed the stress-induced increase in plasma corticosterone levels in defensive withdrawal at doses of 3.0 mg/kg and higher. These doses also resulted in levels of DMP904 in plasma similar to (for anxiolytic-like effects) or 4-fold higher (for effects on the HPA axis) than the in vitro IC(50) value for binding affinity at CRF(1) receptors and greater than 50% occupancy of CRF(1) receptors. Unlike alprazolam, DMP904 did not produce sedation, ataxia, or chlordiazepoxide-like subjective effects (as measured by locomotor activity, rotorod performance, and chlordiazepoxide discrimination assays, respectively) at doses at least 3-fold higher than anxiolytic-like doses. In conclusion, anxiolytic-like effects and effects on the stress-activated HPA axis of DMP904 in the defensive withdrawal model of anxiety required 50% or greater occupancy of central CRF(1) receptors. This level of CRF(1) receptor occupancy resulted in fewer motoric side effects compared with classic benzodiazepines. PMID- 14742751 TI - Effect of atorvastatin on cyclosporine pharmacokinetics in liver transplant recipients. AB - BACKGROUND: The development of hyperlipidemia after liver transplant is frequently treated with hydroxymethylglutaryl coenzyme A reductase inhibitors (statins) such as atorvastatin. As atorvastatin and the primary immunosuppressant drug, cyclosporine, are metabolized by the same pathway, there is the potential for an interaction. OBJECTIVE: To determine the effect of atorvastatin on cyclosporine pharmacokinetics in liver transplant recipients. METHODS: Six stable, long-term adult liver transplant recipients from a single center who developed posttransplant dyslipidemia were recruited to participate in a 14-day, open-label study of atorvastatin 10 mg/d coadministered with standard posttransplant immunosuppression using constant oral doses of cyclosporine and corticosteroids. A 10-point pharmacokinetic profile was performed prior to and on day 14 after commencement of atorvastatin therapy. Cyclosporine concentrations were measured by HPLC-electrospray-tandem mass spectrometry. The AUC was calculated by the linear trapezoidal rule, with other parameters determined by visual inspection. RESULTS: Atorvastatin coadministration increased the cyclosporine AUC by 9% (range 0-20.6%; 3018 vs 3290 ng.h/mL; p = 0.04). No significant change was evident for other cyclosporine pharmacokinetic parameters. Total cholesterol and low-density lipoprotein cholesterol levels were significantly lower on day 14 than at baseline (p < 0.02). One patient developed a twofold increase in transaminases after 2 weeks of atorvastatin therapy, but no other clinical or biochemical adverse events were recorded. CONCLUSIONS: Atorvastatin coadministration increases the cyclosporine AUC by approximately 10% in stable liver transplant recipients. This change in systemic exposure to cyclosporine is of questionable clinical significance. Atorvastatin is effective in reducing cholesterol levels in liver transplant recipients. PMID- 14742752 TI - Ethnic differences in antihypertensive medication use in the elderly. AB - BACKGROUND: Determining the optimal treatment for hypertension in very old patients requires better understanding of interethnic differences in patterns and predictors of antihypertensive drug use in this population. OBJECTIVE: To investigate interethnic variations in antihypertensive drug use in a tri-ethnic sample of community-dwelling adults aged > or =77 years. METHODS: We performed a cross-sectional study of non-Hispanic white, black, and Hispanic adults > or =77 years old residing in Galveston County, TX. In-home interviews in 1997 and 1998 assessed blood pressures and antihypertensive medication use in 281 subjects who reported having hypertension or who had a systolic blood pressure > or =140 mm Hg and/or diastolic blood pressure > or =90 mm Hg. RESULTS: Of the population evaluated, 62.9% of non-Hispanic whites, 60.2% of blacks, and 45.2% of Hispanics with hypertension were on antihypertensive medications (p < 0.027 across the ethnic groups). After adjusting for age, gender, years of education, household income, Medicaid insurance, number of physician visits, and cognitive function, Hispanic ethnicity, unlike black ethnicity, continued to be significantly associated with lower use of antihypertensive drugs compared with non-Hispanic whites (OR 0.41; 95% CI 0.19 to 0.90). Characteristics associated with the lower use of antihypertensive drugs included older age and low income in whites, poor cognition and infrequent physician visits in blacks, and lack of Medicaid insurance in Hispanics. CONCLUSIONS: In the elderly, Hispanic ethnicity, unlike black ethnicity, is significantly associated with lower use of antihypertensive drug therapy compared with non-Hispanic white ethnicity, adjusting for relevant sociodemographic and health factors. PMID- 14742753 TI - Tracking trends in secondary stroke prevention strategies. AB - BACKGROUND: Stroke is defined as a rapid onset of isolated neurologic dysfunction and is the leading cause of disability in adults, as well as the third-leading cause of death in the US. Nearly 600,000 cases of stroke are reported annually. OBJECTIVE: To quantify the impact of emerging evidence on the use of secondary stroke prevention strategies in patients discharged from a specialized stroke center in 2002 compared with those discharged in 2000. METHODS: Using a retrospective approach, data were collected on the first 100 patients cared for by the stroke service from July 1, 2000, to December 31, 2000. Using a prospective approach, the goal was to enroll 100 patients with a diagnosis of ischemic stroke and telephone the patients 1 and 3 months after discharge to determine patient adherence and persistence rate of medications. This is an interim report of the first 20 patients enrolled. RESULTS: Both studies resulted in populations of 55% women and 45% men. The most common risk factor was hypertension, with 59% on admission in 2000 and 75% in 2002. Use of antithrombotics (98% in 2000 to 100% in 2002) and antihypertensives (67% in 2000 to 90% in 2002) was high in both studies and not significantly different. The 2 most significant changes in prescribing were increased use of statins (13% in 2000 to 50% in 2002, p = 0.0007) and combination clopidogrel and aspirin use (0% in 2000 to 20% in 2002, p = 0.001). At 3 months, 66.7% of patients were on the same antithrombotic medication as at discharge. CONCLUSIONS: The dedicated stroke service was able to effectively translate emerging evidence and guidelines into practice and significantly change the use of prescribed medications for secondary stroke prevention. PMID- 14742754 TI - Herbals and asthma: usage patterns among a border population. AB - BACKGROUND: The use of herbal products (HP) is rising in the US. Higher rates of HP use have been documented in the US/Mexico border population, as well as increasing concerns about herbal-related adverse events. OBJECTIVE: To evaluate the prevalence of HP use in adult asthmatic patients requiring hospitalization and the frequency of HP documentation in medical records. METHODS: We conducted a retrospective chart review of admissions for asthma to determine the frequency of HP documentation. Additionally, during a 12-month period, a bilingual interviewer conducted prospective, semistructured interviews with patients with asthma exacerbations to record data on HPs used specifically for the treatment of asthma. RESULTS: A total of 67 cases were chart-reviewed retrospectively; 60 patients were interviewed prospectively. We found no documentation of HP use by chart review, while prospective interviews showed that 42% of patients reported using HPs for the treatment of asthma. The most common HPs used were oregano 28%, chamomile 20%, garlic 16%, eucalyptus 12%, and lime 12%. Ten patients reported taking an HP that could potentially exacerbate their asthma and 18 patients reported using an HP that could interact with other medications or cause other types of adverse events. CONCLUSIONS: An obvious lack of documentation for HP use was observed in the medical records reviewed. Because a number of HPs that are commonly used by residents along the border can interact with antiasthmatic agents and/or result in compromised asthma control, questions about HP use should be included in routine history taking. PMID- 14742755 TI - Involvement of immunization-certified pharmacists with immunization activities. AB - BACKGROUND: Immunization certification courses allow pharmacists to directly administer vaccines to their patients. However, the demographics and level of immunization involvement of immunization-certified pharmacists compared with those noncertified are unknown. OBJECTIVE: To document the demographics, professional activities, and job satisfaction of immunization-certified pharmacists compared with pharmacists not certified for immunization. METHODS: In a cross-sectional pilot study, immunization-certified pharmacists were compared with noncertified pharmacists via a postal-mailed questionnaire. The questionnaire consisted of demographic and practice site characteristics, involvement in immunization services, and a job satisfaction survey. RESULTS: Response rates were 48% (n = 101) and 36% (n = 158) for immunization-certified and noncertified pharmacists, respectively. Significantly more certified pharmacists were involved in immunizations (99% vs 24%; p < 0.001). Desire to improve the health care of the public and personal satisfaction were important factors that encouraged pharmacists to become certified to administer vaccines. Seventy-four percent of immunization-certified pharmacists directly administered the vaccines, primarily influenza (96%), pneumococcal (77%), hepatitis (55%), and diphtheria, pertussis, tetanus (19%). Adequate training, time, support from management and staff, and liability coverage were important factors that allowed pharmacists to incorporate immunizations into their practice. No significant differences in job satisfaction were observed between immunization-certified and noncertified pharmacists. CONCLUSIONS: Immunization-certified pharmacists are using their skills to administer vaccines to patients within their communities. Efforts to increase the number of these pharmacists throughout the US should be undertaken. PMID- 14742756 TI - Rhabdomyolysis and renal failure associated with gemfibrozil monotherapy. AB - OBJECTIVE: To report a case of rhabdomyolysis and acute renal failure associated with gemfibrozil monotherapy of hyperlipidemia. CASE SUMMARY: A 30-year-old white man with hypertension, type 1 diabetes mellitus, and hyperlipidemia was hospitalized due to myalgias, nausea, and vomiting that began after he started working as a jackhammer operator 4 days previously. His medications were lisinopril, aspirin, insulin, and gemfibrozil. Creatine kinase and creatinine, which previously had been mildly elevated and normal, respectively, were markedly elevated, consistent with rhabdomyolysis with acute renal failure. DISCUSSION: As of December 8, 2003, this is the only report of a patient with normal baseline creatinine level who developed rhabdomyolysis with acute renal failure associated with gemfibrozil monotherapy. Strenuous exertion, hypovolemia, and lisinopril use may have contributed to the severity of illness. An objective causality assessment revealed that an adverse drug reaction to gemfibrozil was possible. CONCLUSIONS: Gemfibrozil monotherapy of hyperlipidemia may predispose to rhabdomyolysis with acute renal failure. Patients using gemfibrozil should be cautioned regarding strenuous exertion, dehydration, and the need for prompt evaluation of myalgias. PMID- 14742757 TI - Possible gatifloxacin-induced seizure. AB - OBJECTIVE: To report a case of seizure activity following gatifloxacin administration during treatment of urosepsis in an intensive care unit. CASE SUMMARY: An 86-year-old African American male nursing home resident with a history of hypertension and cerebrovascular accident with aphasia was initiated on gatifloxacin therapy for treatment of urosepsis. After 2 intravenous doses (400 mg followed 24 h later with 200 mg), he developed 2 generalized seizures lasting <2 minutes each. Gatifloxacin was discontinued, and the patient was switched to ceftazidime. Phenytoin was initiated, and no recurrent seizures were witnessed. DISCUSSION: Fluoroquinolones have been shown to induce seizures or decrease seizure threshold. The majority of reports include older-generation fluoroquinolones such as ofloxacin. The incidence of seizures associated with fluoroquinolones varies among the individual agents and is relatively rare. Risk factors include increased age, compromised renal function, history of seizures, and coadministration with theophylline or nonsteroidal antiinflammatory drugs. Application of an objective causality scale indicates the seizures in our patient were possibly associated with gatifloxacin. CONCLUSIONS: This case of gatifloxacin-induced seizures may reflect a class-effect phenomenon versus an effect caused by an individual fluoroquinolone. Caution is warranted when using these agents in patients at risk for drug-induced seizures. PMID- 14742758 TI - Severe dermatologic reactions at multiple sites after paclitaxel administration. AB - OBJECTIVE: To report a case of a severe dermatologic reaction at the infusion site, as well as at multiple distant sites, after paclitaxel administration. CASE SUMMARY: A 53-year-old white male with cancer of the base of the tongue was treated with paclitaxel 100 mg/m2 infused over 3 hours and carboplatin dosed at an AUC of 6 mg/mL x min infused over 30 minutes via a peripheral vein on the left arm. After 4 doses of paclitaxel, the patient developed erythematous patches on both forearms and both thighs. The lesions on the left arm worsened into a necrotic ulcer, exposing underlying tissues. All of the lesions healed after paclitaxel was withheld from subsequent cycles. Based on the Naranjo probability scale, it is probable that paclitaxel was the causative agent for these lesions. DISCUSSION: Most of the previously reported paclitaxel-induced dermatologic reactions occurred following extravasation. There are few reports of generalized dermatologic rashes. These rashes are thought to be due to delayed hypersensitivity either to paclitaxel or the solubilizing agents. Our patient had dermatologic toxicity at the infusion site, as well as at multiple other sites, that developed about 6 weeks after the first paclitaxel infusion. CONCLUSIONS: Generalized dermatologic toxicity after paclitaxel administration is uncommon and seems to be immunologically mediated. This toxicity could be secondary to paclitaxel or the additives. The use of paclitaxel is increasing in various malignancies. Newer formulations are under development. Until these formulations are available, clinicians should report any dermatologic reactions in order to identify the cause of toxicity and develop safer administration technique. PMID- 14742760 TI - Low-molecular-weight heparin-induced thrombocytopenia in a child. AB - OBJECTIVE: To report a case of probable acute venous thrombosis caused by heparin induced thrombocytopenia (HIT) in a pediatric patient with a normal platelet count after prolonged enoxaparin therapy. CASE SUMMARY: An 11-year-old African American female with Crohn's disease developed extensive vena cava thrombosis. Her deep vein thrombosis (DVT) was treated with intravenous unfractionated heparin followed by extended outpatient warfarin therapy. Four months later, the warfarin was stopped and subcutaneous enoxaparin 1.5 mg/kg once daily was substituted prior to an elective colonoscopy. She was readmitted 6 weeks later with acute DVT with a platelet count of 233 x 10(3)/mm3, significantly lower than the count of 550-700 x 10(3)/mm3 5 months previously and the count of 433 x 10(3)/mm3 3 months earlier. An enzyme-linked immunosorbent assay for heparin platelet factor 4 antibodies was strongly positive and a d-dimer was elevated at 2.9 mg/L (normal <1.5). She was treated with lepirudin followed by warfarin when repeat d-dimer on day 3 was normal. An ultrasound at that time showed no clot extension, and the platelet count had risen to >300 x 10(3)/mm3. Over the next 4 months, there was no further thrombosis. DISCUSSION: HIT appears to be rare in the pediatric population, and only a few cases treated with a direct thrombin inhibitor have been reported. This is the first case report to our knowledge of a pediatric patient developing HIT secondary to enoxaparin. An interesting feature of this case is the development of HIT in the face of a normal platelet count, which is rare but has been reported in adults. CONCLUSIONS: Pediatric patients receiving low-molecular-weight heparin are still at risk for developing HIT. Treatment of HIT should involve the initial use of a direct thrombin inhibitor to manage thrombosis until the platelet count returns to higher values. Once the platelet count returns, warfarin can be used for long-term thrombosis management. PMID- 14742759 TI - Vascular neurotoxicity following chemotherapy with cisplatin, ifosfamide, and etoposide. AB - OBJECTIVE: To report a case of acute central nervous system (CNS) toxicity with multiple hemorrhages restricted to the corpus callosum associated with combination therapy of cisplatin, ifosfamide, and etoposide. CASE SUMMARY: A 38 year-old white man with a testicular germ cell tumor received a cisplatin-based chemotherapy consisting of cisplatin 45 mg (20 mg/m2), etoposide 570 mg (250 mg/m2), and ifosfamide 4600 mg (2000 mg/m2) given on 5 consecutive days during each course. After the first course of chemotherapy, the patient appeared to be neuropsychologically impaired with episodes of decreased alertness and features of a depressive syndrome. He became severely diminished in mental function, orientation, and psychomotor activity after a second course of treatment. In addition, he showed transient urinary incontinence. Motor and sensory deficits could not be detected. Magnetic resonance imaging demonstrated multiple hemorrhages restricted to the corpus callosum. An objective causality assessment revealed that an adverse drug reaction was probable. DISCUSSION: Neurotoxicity has been associated with the administration of various antineoplastic agents. In particular, cisplatin and ifosfamide can cause both acute and delayed CNS toxicity. While ifosfamide neurotoxicity has been predominantly associated with neuropsychological impairment without evidence of structural abnormalities in neuroimaging studies, cisplatin has been shown to cause cerebrovascular complications. Various pathophysiologic conditions may contribute to these complications including thrombosis secondary to vascular endothelial injury or thromboembolic events. To our knowledge, as of December 2, 2003, vascular lesions restricted to the corpus callosum have not been reported as a complication of cisplatin- or ifosfamide-based chemotherapy. CONCLUSIONS: Clinicians should be aware of the potential neurovascular adverse effects of cisplatin-based protocols. This is especially true in patients with subtle neurologic or neuropsychological symptoms. Chemotherapy-induced neurotoxicity should be considered in the differential diagnosis. PMID- 14742761 TI - Warfarin initiation and monitoring with clotting factors II, VII, and X. AB - OBJECTIVE: To report a case of a patient with antiphospholipid antibody syndrome and multiple thromboses who developed heparin-induced thrombocytopenia (HIT) and subsequent international normalized ratio (INR) prolongation possibly due to antiphospholipid antibodies. CASE SUMMARY: A 56-year-old white woman with a history of antiphospholipid antibody syndrome and thrombosis taking chronic warfarin was admitted for gastrointestinal concerns and found to have an INR >14. Warfarin was discontinued, vitamin K was administered, and a heparin infusion was initiated. Over the next 2 days, thrombocytopenia, hypotension, tachycardia, hyponatremia, and progressive abdominal pain developed. Upon transfer to a tertiary care center, HIT was diagnosed, and a lepirudin infusion was initiated. Subsequently, a sudden elevation of the INR occurred (>14) with low prothrombin (factor II) activity. After INR values declined to 2-3, warfarin was reinitiated with dosing adjusted using factor X and II activity levels. Clotting factors II and X activities were measured to monitor long-term warfarin therapy, with no evidence of complications after 7 months. DISCUSSION: Typically, the INR is used to assess the intensity of anticoagulation. The INR value represents the reduction of clotting factors II, VII, and X. In rare circumstances, an independent inhibitor or interfering substance can interfere with the process of measuring the INR. In such situations, an alternative approach can be direct measurement of clotting factor concentrations. CONCLUSIONS: Factor II and/or factor X activity levels provided an alternative means for measuring the anticoagulant effects of warfarin in the presence of a significant inhibitor (antiphospholipid antibodies) that biased the INR measurements. PMID- 14742762 TI - Ginger-associated overanticoagulation by phenprocoumon. AB - OBJECTIVE: To report a case of ginger-phenprocoumon interaction resulting in an elevated international normalized ratio (INR) and epistaxis. CASE SUMMARY: A 76 year-old white European woman on long-term phenprocoumon therapy with an INR within the therapeutic range began using ginger products. Several weeks later, she developed an elevated INR up to 10 and epistaxis. The INR returned to the normal range after ginger was stopped and vitamin K1 was given. DISCUSSION: There have been a number of investigations resulting in conflicting opinions on the effect of ginger on hemostasis, specifically, platelet inhibition. Nevertheless, based on these investigations, recommendations have been issued to refrain from ingesting ginger and other herbals like garlic or ginkgo biloba in situations where bleeding may be critical. An objective causality assessment revealed that the adverse drug event as a result of the phenprocoumon and ginger interaction was probable. CONCLUSIONS: As of writing, this was the first case report that may support an interaction between an oral anticoagulant and ginger together with a brief review of the literature on ginger and hemostasis. As this interaction was observed only by chance, this case highlights the importance of self-control of anticoagulation with coumarins particularly for the detection of unknown interactions. PMID- 14742763 TI - Acenocoumarol-induced Henoch-Schonlein purpura. AB - OBJECTIVE: To report a probable case of Henoch-Schonlein purpura associated with acenocoumarol therapy. CASE SUMMARY: A 76-year-old white woman was prescribed acenocoumarol for chronic atrial fibrillation. Two months after starting therapy, the patient came to our hospital's emergency department because of abdominal pain associated with vomiting. Physical examination revealed multiple round, confluent, purpuric lesions with some vesicles and an area of residual pigmentation. Lesions were present predominantly on the legs and gluteus, and also on the abdomen and arms. Skin biopsy of the lesions was compatible with leukocytoclastic vasculitis with deposition of immunoglobulin A. An upper intestinal endoscopy was done and identified purpuric mucosal lesions in the fundus, body, and antrum of the stomach and the duodenal bulb. Renal function was not affected, although proteinuria (1.26 g/day) was found and microscopic hematuria was observed. DISCUSSION: The most likely cause of the Henoch-Schonlein purpura in this case was considered to be acenocoumarol because of the close temporal relationship between exposure to the drug and onset of symptoms, as well as the rapid resolution of the symptoms and signs after acenocoumarol was discontinued. The oral anticoagulant was the only identifiable precipitant that the patient encountered before the Henoch-Schonlein purpura developed. An objective causality assessment revealed that the adverse drug event was probable. CONCLUSIONS: This case report illustrates a probable association between Henoch Schonlein purpura and acenocoumarol. As of December 2003, this reaction had not been previously reported. Clinicians should be aware of this potential adverse effect of a widely used drug. PMID- 14742764 TI - Metabolic bone disease after chronic antacid administration in an infant. AB - OBJECTIVE: To describe a case of hypophosphatemia and metabolic bone disease (MBD) due to aluminum-containing antacids. CASE SUMMARY: An 8-month-old white boy was brought to the emergency department due to irritability and pain with movement. Upon examination, the infant was irritable, unable to bear weight, had palpable clavicular bony lesions, point tenderness of the hips, and poor head control. The infant had lost several developmental milestones over the past 4 months (eg, ability to roll over) and had decreased appetite and minimal weight gain. Skeletal survey revealed multiple rib fractures, osteoporosis, and Ricketts. Hypophosphatemia (2.3 mg/dL; normal 3.2-6.3) and an elevated serum aluminum level (14 microg/L, normal 0-9) were noted. Past medical history was positive for gastroesophageal reflux. He had been started on ranitidine and aluminum hydroxide (1/2 teaspoonful per 6-ounce bottle) at 2 months of age. The infant's formula contained elevated aluminum levels. Further investigation showed that 1/2 tablespoonful instead of 1/2 teaspoonful of antacid had been added to each 6-ounce formula bottle for the prior 6 months; only 1 month of antacid therapy had been recommended. An objective causality assessment revealed a probable adverse drug event. DISCUSSION: Phosphate-binding substances such as aluminum-containing antacids can bind large amounts of phosphorus, causing hypophosphatemia and MBD. CONCLUSIONS: We report this case to increase awareness of the risk of hypophosphatemia and MBD (likely aluminum related) with use of over-the-counter aluminum-containing antacids in pediatrics, as well as the hazards of prescribing doses in "spoonful" units. PMID- 14742765 TI - Serotonin syndrome with elevated paroxetine concentrations. AB - OBJECTIVE: To describe a case of serotonin syndrome due to paroxetine and ethanol. CASE SUMMARY: A 57-year-old white man was brought to the emergency department one day after ingesting paroxetine 3600 mg and a pint of hard liquor. He denied the use of any other drug or herbal products and regular use of alcohol. Upon arrival to the hospital, vital signs were blood pressure 188/103 mm Hg, heart rate 114 beats/min, respiratory rate 28 breaths/min, temperature 36.8 degrees C, and O2 saturation 96% on room air. Findings on physical examination included dilated pupils, facial flushing, diaphoresis, shivering, myoclonic jerks, tremors, and hyperreflexia. A tentative diagnosis of serotonin syndrome was made. Initially, cyproheptadine 8 mg was administered orally with no observable effect. An additional 12 mg was given in 3 doses over 24 hours. Symptoms abated slowly over the next 6 days, during which a thorough evaluation failed to reveal any other potential causes for the patient's condition. Serum paroxetine concentrations at 27.5 and 40 hours after ingestion were 1800 and 1600 ng/mL, respectively (normal 20-200 ng/mL). DISCUSSION: Serotonin syndrome is rarely reported in patients taking only one serotonergic medication. Although serum paroxetine concentrations have not been shown to correlate with efficacy or toxicity, our patient's serum paroxetine concentration was 9 times the upper end of the therapeutic range. Cyproheptadine, which has been suggested as a therapy, did not appear beneficial in this patient. Use of the Naranjo probability scale indicated a probable relationship between the serotonin syndrome and the overdose of paroxetine taken by this patient. CONCLUSIONS: More studies are needed to better assess the role of cyproheptadine and other serotonin antagonists in the management of the serotonin syndrome. Regardless of the use of cyproheptadine or other agents, attention should be paid to fluid status, decontamination, and management of hyperthermia, agitation, and seizures. PMID- 14742766 TI - Transfer of risperidone and 9-hydroxyrisperidone into human milk. AB - OBJECTIVE: To quantify the transfer of risperidone and its active metabolite 9 hydroxyrisperidone into breast milk, estimate the amount the infant receives, measure infant plasma concentrations, and clinically assess the safety of breast feeding during maternal risperidone administration. case summaries: The transfer of risperidone and 9-hydroxyrisperidone into milk was studied in 2 breast-feeding women and one woman with risperidone-induced galactorrhea. Plasma samples were available from 2 of the women and from both breast-fed infants. The milk/plasma concentration ratio determined in 2 women was <0.5 for both compounds. The calculated relative infant "doses" were 2.3%, 2.8%, and 4.7% (as risperidone equivalents) of the maternal weight-adjusted doses. Risperidone and 9 hydroxyrisperidone were not detected in the plasma of the 2 breast-fed infants studied, and no adverse effects were noted. DISCUSSION: Risperidone therapy is sometimes necessary in breast-feeding women, raising the issue of safety in the exposed infants. Our study shows that the relative infant dose is lower than the arbitrary 10% level of concern. The data provide clear guidance on infant exposure for the cases presented. CONCLUSIONS: Maternal risperidone therapy is unlikely to be a significant hazard for the breast-fed infant in the short term. Nevertheless, decisions on whether a woman may breast-feed should be made as an individual risk-benefit analysis. PMID- 14742767 TI - The metabolic syndrome: pathophysiology, clinical relevance, and use of niacin. AB - OBJECTIVE: To review the pathophysiology and clinical relevance for using niacin to treat the metabolic syndrome. DATA SOURCES: Primary articles were identified through a MEDLINE search (1966-January 2003), and recommendations for treatment were obtained from the National Cholesterol Education Program Adult Treatment Panel (NCEP-ATP) III guidelines. STUDY SELECTION AND DATA EXTRACTION: Published studies showing the effects of the metabolic syndrome, atherogenic dyslipidemia, and niacin were evaluated and reviewed. DATA SYNTHESIS: The metabolic syndrome is a highly prevalent condition that affects 24% of American adults and significantly increases the risk of coronary heart disease (CHD). Most patients with metabolic syndrome have atherogenic dyslipidemia characterized by elevated triglycerides, low high-density-lipoprotein cholesterol (HDL-C), and small, dense low-density-lipoprotein cholesterol (LDL-C) particles. The NCEP-ATP III identifies patients with the metabolic syndrome as candidates for intensified therapy. Lifestyle modifications and drug therapy are recommended. Niacin represents a good option for treating the triad of lipid abnormalities seen in the metabolic syndrome because it raises HDL-C, lowers triglycerides, and increases LDL-C particle size. CONCLUSIONS: Treatment of the metabolic syndrome is recommended by NCEP-ATP III to further reduce CHD risk after the LDL-C target has been met. Prospective clinical studies are needed to define the impact of niacin and other lipid-modifying agents on CHD morbidity and mortality in patients with the metabolic syndrome. PMID- 14742768 TI - Beta-blocker underuse in secondary prevention of myocardial infarction. AB - OBJECTIVE: To review the clinical benefits of beta-blockers as secondary prevention following a myocardial infarction (MI) and to address the reasons that clinicians are reluctant to use beta-blockers in specific patient populations. DATA SOURCES: MEDLINE was searched for articles published from January 1966 to October 2002. Relevant studies were identified by systematic searches of the literature for all reported studies of associations between beta-blocker underuse and secondary prevention of MI. Additional studies were identified by a hand search of references of original or review articles. STUDY SELECTION AND DATA EXTRACTION: English-language human studies were selected and analyzed. DATA SYNTHESIS: Associations were observed in studies of beta-blocker use as secondary prevention of MI. A lower rate of beta-blocker treatment occurred in older patients and in patients with comorbid conditions such as diabetes, heart failure, chronic obstructive pulmonary disease, asthma, and peripheral arterial disease. In addition, underuse was attributed to the perception of high rates of adverse events associated with beta-blockers. beta-Blocker use as secondary prevention of an MI can lead to a 19-48% decrease in mortality and up to a 28% decrease in reinfarction rates. Nonetheless, beta-blockers are significantly underused in many patient populations due to concomitant disease states. Due to their normal physiologic deterioration, the elderly are at an increased risk of low cardiac output and bradycardia when given a beta-blocker; therefore, they should be started on a low dose that is then slowly titrated. In diabetic patients, beta-blockers can impair glucose control leading to hypoglycemia; therefore, post-MI diabetic patients must routinely monitor their blood glucose levels. In patients with decompensated heart failure, beta-blocker use can lead to further cardiac depression, but lower oral starting doses with slow titration can reduce this risk. beta-Blockers can induce bronchospasm in patients with chronic obstructive pulmonary disease or asthma, but cardioselective beta blockers and appropriate use of medications such as albuterol can minimize these effects. Finally, in patients with peripheral arterial disease, with the exception of hypertensive patients with Reynaud's phenomenon, beta-blockers can be used safely. The only absolute contraindications to beta-blockers are severe bradycardia, preexisting sick sinus syndrome, second- and third-degree atrioventricular block, severe left ventricular dysfunction, active peripheral vascular disease with rest ischemia, or reactive airway disease so severe that airway support is required. CONCLUSIONS: Overall, the cardiovascular benefits of beta-blockers as secondary prevention of MI significantly outweigh the risks associated with their use. PMID- 14742769 TI - Teriparatide for severe osteoporosis. AB - OBJECTIVE: To review the pharmacology, toxicology, pharmacokinetics, pharmacodynamics, efficacy, safety, therapeutic controversies, administration, patient counseling, and formulary recommendations for teriparatide (rDNA origin). DATA SOURCES: A MEDLINE search (1966-May 2003) of articles using the key words parathyroid hormone and osteoporosis, parathyroid hormone and fracture, and teriparatide was conducted to identify relevant literature in the English language. Additional references were obtained from bibliographies of those articles. Some clinical trial data not yet published were obtained from the manufacturer. STUDY SELECTION AND DATA EXTRACTION: All articles obtained from the data sources were reviewed; all data deemed relevant were included. DATA SYNTHESIS: Teriparatide, recombinant human parathyroid hormone (1-34) [rhPTH (1 34)], is the first anabolic agent to treat postmenopausal women with osteoporosis and men with idiopathic or hypogonadal osteoporosis who are at high risk for osteoporotic fracture. Daily subcutaneous injections of teriparatide significantly increase both spine and hip bone-mineral density (BMD) while decreasing the incidence of fractures in both women and men. Common adverse effects noted with teriparatide use were nausea, headache, dizziness, and arthralgias. An increased incidence of osteosarcoma in rats during preclinical trials with teriparatide led to a black box warning for the drug. CONCLUSIONS: Teriparatide substantially increases spine and hip BMD and may offer additional benefits to patients with severe osteoporosis. Clinical trials comparing teriparatide with other available agents to treat osteoporosis are needed to more clearly define its place in therapy. Long-term safety and efficacy are not known. PMID- 14742770 TI - Measurement, correlates, and health outcomes of medication adherence among seniors. AB - OBJECTIVE: To provide a comprehensive review of the literature on the measurement, correlates, and health outcomes of medication adherence among community-dwelling older adults. DATA SOURCES: Searches of MEDLINE, PubMed, and International Pharmaceutical Abstracts databases for English-language literature (1966-December 2002) were conducted using one or more of the following terms: elderly, adherence/nonadherence, compliance/noncompliance, medication/drug, methodology/measurement, and hospitalization. STUDY SELECTION AND DATA EXTRACTION: From the above search, studies of medication adherence in community dwelling seniors were selected for review along with relevant publications from the reference lists of articles identified in the initial database search. DATA SYNTHESIS: Although several methods are available for the assessment of adherence, accurate measurement continues to be difficult. The available evidence suggests that polypharmacy and poor patient-healthcare provider relationships (including the use of multiple providers) may be major determinants of nonadherence among older persons, with the impact of most sociodemographic factors being negligible. There is little consensus regarding other determinants of nonadherence. Relatively few high-quality investigations have examined the associations between nonadherence and subsequent health outcomes. Available data provide some support for increased health risks with nonadherence. However, interventions to improve adherence have seldom demonstrated positive effects on health outcomes. CONCLUSIONS: There are few empirical data to support a simple systematic descriptor of the nonadherent patient. The inconsistencies across studies may be attributable, in part, to the inherent difficulties involved in the measurement of a behavioral risk factor such as nonadherence. Future research in this area would be strengthened by incorporation of detailed assessments of patient-reported reasons for nonadherence, the appropriateness of drug regimens, and the effect of nonadherence on health outcomes. PMID- 14742771 TI - Selection of atypical antipsychotics for the management of schizophrenia. AB - OBJECTIVE: To review the evidence for selecting one atypical antipsychotic agent over another for management of schizophrenia. DATA SOURCES: A literature search of MEDLINE (1966-June 2003), EMBASE (1998-June 2003), and the Cochrane Library was conducted using the following terms: schizophrenia, quetiapine, ziprasidone, olanzapine, aripiprazole, and risperidone. Bibliographies of relevant articles were hand-searched for additional references. STUDY SELECTION AND DATA EXTRACTION: Prospective, randomized, blinded trials and meta-analyses that directly or indirectly compared > or =2 atypical antipsychotic agents in the management of schizophrenia are included in this review. Studies comparing an atypical agent with clozapine are not included. DATA SYNTHESIS: A small number of prospective, randomized, blinded trials that compare efficacy and tolerability of olanzapine and risperidone have been published. These trials did not reveal clinically meaningful differences in efficacy but did confirm that their adverse effect profiles are slightly different (more weight gain with olanzapine and more extrapyramidal reactions with risperidone). Direct comparisons between other atypical antipsychotics are not available. Systematic reviews (indirect comparisons) of placebo-controlled or traditional antipsychotic-controlled trials suggest similar efficacy for quetiapine, olanzapine, and risperidone when placebo is the comparator and inferior efficacy of quetiapine compared to olanzapine and risperidone when haloperidol is the comparator. The few available economic analyses are difficult to interpret in light of current practice. CONCLUSIONS: Additional randomized, blinded clinical trials directly comparing efficacy, tolerability, and cost-effectiveness are needed to confirm the proposed differences among atypical antipsychotic agents before recommendations can be made with confidence. PMID- 14742772 TI - Purified poloxamer 188 for sickle cell vaso-occlusive crisis. AB - OBJECTIVE: To review available literature on the pharmacology, pharmacokinetics, efficacy, toxicology, adverse effects, drug interactions, and dosage guidelines for purified poloxamer 188, a product in Phase III trials. DATA SOURCES: Reviewers searched the following databases for English-language studies: MEDLINE (1966-November 2003), International Pharmaceutical Abstracts (1970-November 2003), and the Cochrane Library Database (3rd quarter 2003). Key search terms included purified poloxamer 188, Flocor, CRL-5861, poloxamer 188, RheothRx, and pluronic F-68. STUDY SELECTION AND DATA EXTRACTION: Data on efficacy, adverse effects, and pharmacokinetics were obtained from randomized, open-label, and blinded clinical trials. Toxicology data were obtained from unpublished studies with purified poloxamer 188 and from available data on poloxamer 188 (nonpurified form). DATA SYNTHESIS: Purified poloxamer 188 is a highly purified form of the nonionic block copolymer poloxamer 188. It lowers blood viscosity, decreases red blood cell (RBC) aggregation, and decreases friction between RBCs and vessel walls to increase microvascular blood flow and decrease cell injury. In clinical trials, purified poloxamer 188 demonstrated safety, but little efficacy for the treatment of sickle cell vaso-occlusive crisis. Increased efficacy has been shown in patients on concurrent hydroxyurea therapy and those <15 years of age. CONCLUSIONS: Purified poloxamer 188 represents a new approach to the management of the sickle cell vaso-occlusive crisis. Children and patients on hydroxyurea may benefit most from purified poloxamer 188 therapy. Further studies are needed to confirm its efficacy and to determine whether the drug decreases sickle cell disease severity and complications. PMID- 14742773 TI - Urinary-based ovulation and pregnancy: point-of-care testing. AB - OBJECTIVE: To review the literature concerning ovulation prediction devices and pregnancy detection tests for home use. DATA SOURCES: Articles were identified through searches of the MEDLINE (1966-May 2003), EMBASE (1980-May 2003), and International Pharmaceutical Abstracts (1970-May 2003) databases using the key words ovulation, ovulation detection, pregnancy test, diagnostic reagent kit, and diagnostic test. Additional references were located through review of the bibliographies of the articles found in the literature search. Searches were not limited by time restriction, language, or use of human or animal subjects. STUDY SELECTION AND DATA EXTRACTION: Review articles, textbook chapters, and experimental and observational studies on home use ovulation and pregnancy tests were selected. DATA SYNTHESIS: Luteinizing hormone (LH)-based ovulation tests have demonstrated accurate and superior ovulation detection when compared to basal body temperature charting, calendar calculation, salivary ferning, or observation of vaginal or cervical discharge changes. Systems using LH and estrone-3-glucuronide (E3G) have also demonstrated accurate detection of the fertile period. Literature evaluating home use of pregnancy tests has demonstrated accurate use by lay persons. CONCLUSIONS: Urinary-based ovulation prediction and pregnancy detection tests available for use by nonprofessionals enable women and couples to take an active role in the family planning process. Numerous products are available at reasonable costs to the consumer. PMID- 14742774 TI - Single versus combined antibiotic therapy for gram-negative infections. AB - OBJECTIVE: To evaluate the available clinical data regarding single versus combination antimicrobial therapy for treatment of gram-negative infections, focusing on the more recent data in predominantly nonneutropenic hosts. In vitro and in vivo data regarding various antimicrobial combinations are also discussed. DATA SOURCES: Clinical trials, review articles, and meta-analyses were identified from a MEDLINE search (1960-July 2003). Special attention was given to clinical outcome trials performed since 1989. Search terms included gram-negative infections, drug synergism, Pseudomonas aeruginosa, monotherapy, combination therapy, carbapenems, beta-lactams, cefepime, aminoglycosides, and fluoroquinolones. DATA SYNTHESIS: Although most of the studies were not randomized, double-blind, or controlled, the most recent literature indicates that monotherapy with agents that are active against isolated organisms, including P. aeruginosa, may be appropriate for most patients. Efficacy outcomes, including mortality, did not significantly differ in most studies comparing single and combination therapies. Some trials suggest that combination therapy may be preferred in neutropenic patients and those with pseudomonal infections. CONCLUSIONS: Hospitalized patients with gram-negative infections are often treated with combination antimicrobial agents; however, some of the recently available data, although limited, suggest that administration of monotherapy is a feasible alternative in certain patient populations. PMID- 14742776 TI - HMG-CoA reductase inhibitors for the prevention of nephropathy. AB - OBJECTIVE: To evaluate the literature to determine whether hydroxymethylglutaryl coenzyme A reductase inhibitors are effective for the prevention of nephropathy. DATA SOURCES: MEDLINE (1966-April 2003) and International Pharmaceutical Abstracts(1970-April 2003), as well as bibliographic searches, were conducted. DATA SYNTHESIS: Although the statins are widely used for the prevention of coronary heart disease, non-lipid-lowering effects are also being investigated, namely their potential role in the prevention of nephropathy. Five trials of the statins used in this manner are reviewed, most of which included patients with dyslipidemias, making it difficult to determine whether the renoprotective effects were independent of the lipid-lowering effects. CONCLUSIONS: Adequate evidence does not currently exist to support the widespread use of statins as alternatives to strategies known to prevent the progression of renal disease, but statins may be used to complement other therapies in patients with additional indications. PMID- 14742775 TI - Trimethoprim/sulfamethoxazole for treatment of severe Staphylococcus aureus infections. AB - OBJECTIVE: To evaluate the role of trimethoprim/sulfamethoxazole (TMP/SMX) as an alternative to vancomycin for the treatment of severe Staphylococcus aureus infections. DATA SOURCES: Clinical literature was accessed through MEDLINE (1966 February 2003) and EMBASE (1980-February 2003). Key search terms included trimethoprim/sulfamethoxazole combination and Staphylococcus aureus. DATA SYNTHESIS: An evaluation of case reports, case series, and clinical studies focusing on the use of TMP/SMX for treatment of severe S. aureus infections was conducted. The majority of the reports indicate that TMP/SMX may be effective for the treatment of infections due to low bacterial burdens of susceptible strains of S. aureus. CONCLUSIONS: In select infections, TMP/SMX may be a useful alternative to vancomycin for treatment of severe S. aureus infections. Additional randomized studies should be conducted comparing this agent with vancomycin and linezolid. PMID- 14742777 TI - Drug control authorities in China. AB - OBJECTIVE: To provide an overview of China's drug control authorities. DATA SOURCES: South Pharmaceutical Economy Institute. DATA SYNTHESIS: China's drug control authorities include the State Food and Drug Administration (SFDA), which primarily oversees drug administration; the State Development and Reform Commission (SDRC), which is responsible for the drug price administration; and the Provincial Administration for Industry and Commerce, cooperating with the Provincial Food and Drug Administration, to regulate drug advertisements. Although the drug control authorities presented here have played important roles in drug administration and have obtained outstanding achievements in the past several years, they are also hindered by their respective problems and try to explore the solutions. CONCLUSIONS: Other countries may confront the problems described here. It is therefore necessary to learn the current situation of drug control authorities in China, as well as the measures being taken to improve the situation. This understanding will aid countries seeking to enter China's pharmaceutical market. PMID- 14742778 TI - Alteplase for central catheter clearance: 1 mg/mL versus 2 mg/2 mL. PMID- 14742779 TI - Low-dose venlafaxine-associated liver toxicity in chronic hepatitis. PMID- 14742780 TI - Verteporfin and myocardial infarction. PMID- 14742781 TI - An unexpected increase of troponin I after perphenazine depot injection. PMID- 14742782 TI - Immediate hypersensitivity to levofloxacin diagnosed through skin prick test. PMID- 14742783 TI - Update: influenza-associated deaths reported among children aged <18 years- United States, 2003-04 influenza season. PMID- 14742785 TI - Inappropriate medication use among frail elderly inpatients. AB - BACKGROUND: Inappropriate prescribing in frail elderly inpatients has not received as much investigation as in frail elderly nursing home patients. OBJECTIVE: To determine the prevalence and predictors of inappropriate prescribing for hospitalized frail elderly patients. METHODS: The study was conducted at 11 Veterans Affairs Medical Centers and involved a sample of 397 frail elderly inpatients. Inappropriate prescribing was measured by physician pharmacist pair's consensus ratings for 10 criteria on the Medication Appropriateness Index (MAI). The MAI ratings generated a weighted score of 0-18 per medication (higher score = more inappropriate) and were summed across medications to achieve a patient score. RESULTS: Overall, 365 (91.9%) patients had > or =1 medications with > or =1 MAI criteria rated as inappropriate. The most common problems involved expensive drugs (70.0%), impractical directions (55.2%), and incorrect dosages (50.9%). The most common drug classes with appropriateness problems were gastric (50.6%), cardiovascular (47.6%), and central nervous system (23.9%). The mean +/- SD MAI score per person was 8.9 +/- 7.6. Stepwise ordinal logistic regression analyses revealed that both the number of prescription (adjusted OR 1.28; 95% CI 1.21 to 1.36) and nonprescription drugs (adjusted OR 1.17; 95% CI 1.06 to 1.29) were related to higher MAI scores. Analyses excluding the number of drugs revealed that the Charlson index (adjusted OR 1.62; 95% CI 1.12 to 2.35) and fair/poor self-rated health (adjusted OR 1.15; 95% CI 1.05 to 1.26) were related to higher MAI scores. CONCLUSIONS: Inappropriate drug prescribing is common for frail elderly veteran inpatients and is related to polypharmacy and specific health status characteristics. PMID- 14742786 TI - High-dose versus standard-dose amoxicillin for acute otitis media. AB - BACKGROUND: Emergence of drug-resistant bacteria has led to a recommendation to use high-dose (HD) amoxicillin (80-90 mg/kg/d) rather than standard-dose (SD) amoxicillin (40-45 mg/kg/d) to treat children with acute otitis media (AOM). OBJECTIVE: To compare the efficacy and tolerability of HD versus SD amoxicillin among children with AOM who were considered at low risk for infection with antibiotic-resistant bacteria. METHODS: A double-blind, randomized, 3-year clinical trial was conducted using participants who met the following criteria: age >3 mo, weight 40 years of age and admitted for congestive heart failure, chronic obstructive pulmonary disease, or respiratory infection were considered appropriate candidates for VTE prophylaxis if they had no documented bleeding risk factors. Patients considered at increased risk of bleeding included those with documented uncontrolled hypertension, thrombocytopenia, coagulopathy, or recent gastrointestinal bleeding. Prescribed regimens were evaluated to determine whether they were consistent with regimens proven in clinical trials to be effective and safe. RESULTS: Thirty-one percent of the patients with established VTE risk factors and no documented risk factors for bleeding were prescribed pharmacologic VTE prophylaxis. An established regimen was prescribed in only 19% of those receiving prophylaxis. CONCLUSIONS: There is significant underutilization of VTE prophylaxis in this patient population. Patients are not adequately assessed for bleeding risk factors, and a portion of prescribed regimens are not those that have been established in the literature. Expert consensus statements recommend that hospitals develop strategies to prevent VTE events in their patients. Strategies to improve patient screening and physicians' prescribing habits are needed. PMID- 14742791 TI - Acute interstitial nephritis due to pantoprazole. AB - OBJECTIVE: To describe what is believed, as of November 4, 2003, to be the first case published in the literature of acute interstitial nephritis (AIN) due to pantoprazole. CASE SUMMARY: A 77-year-old white woman presented to the hospital with elevated serum creatinine, oliguria for the past 24 hours, arthralgia, fatigue, fever, and bilateral flank pain. The patient had initiated treatment with oral pantoprazole 40 mg/d for gastroesophageal reflux 2 months prior to admission. After 5 weeks of therapy, she stopped taking pantoprazole due to general malaise. Upon admission, all home medications, including pantoprazole, were reinitiated based on the patient's medication list. Serum creatinine increased to 6.1 mg/dL on day 4 of admission from a baseline of 1.0 mg/dL. Pantoprazole therapy was promptly discontinued, and prednisone 40 mg/d was initiated. Urinalysis revealed eosinophils, and a subsequent renal biopsy confirmed a diagnosis of AIN. The serum creatinine level gradually declined over 2 weeks, and the patient was discharged home with a serum creatinine level of 1.6 mg/dL. The Naranjo probability scale suggests a highly probable relationship between AIN and pantoprazole therapy in this patient. DISCUSSION: Drug hypersensitivity reactions are the most common cause of AIN. There have been several reported cases of omeprazole-induced AIN. Although there are very few prospective data on the efficacy of treatment of drug-induced AIN, corticosteroids may have a role in recovery of renal function. Prednisone doses of 1 mg/kg/d have been suggested. CONCLUSIONS: Physicians should be aware that drug-induced AIN can be associated with proton-pump inhibitors. Early detection of this rare adverse reaction may prevent acute renal insufficiency. PMID- 14742792 TI - Cushing's syndrome due to interaction between inhaled corticosteroids and itraconazole. AB - OBJECTIVE: To report a case of an interaction between inhaled corticosteroids and itraconazole causing iatrogenic Cushing's syndrome and provide a review of the relevant literature. CASE SUMMARY: A 70-year-old white woman on long-term treatment with high-dose inhaled corticosteroids for asthma was diagnosed as having Scedosporium apiospermum infection of the skin and subcutaneous tissues. As a result, she was treated with itraconazole for 2 months. She subsequently developed Cushing's syndrome due to a probable cytochrome P450-mediated interaction between itraconazole and budesonide. She also had secondary adrenal insufficiency requiring prolonged treatment with replacement hydrocortisone. DISCUSSION: Budesonide is a potent glucocorticoid that is metabolized in the liver by the CYP3A4 isoenzyme to inactive metabolites. Itraconazole is a potent cytochrome P450 inhibitor. It can inhibit the metabolism of oral or inhaled corticosteroids, producing cortisol excess leading to Cushing's syndrome and adrenal insufficiency. An assessment of causality indicated a possible adverse interaction between itraconazole and budesonide. CONCLUSIONS: The combination of itraconazole and inhaled corticosteroids is increasingly being used to treat conditions such as allergic bronchopulmonary aspergillosis. Clinicians need to be aware of the potential for an interaction between such a combination. PMID- 14742793 TI - Fish oil interaction with warfarin. AB - OBJECTIVE: To report a case of elevated international normalized ratio (INR) in a patient taking fish oil and warfarin. CASE SUMMARY: A 67-year-old white woman had been taking warfarin for 1(1/2) years due to recurrent transient ischemic attacks. Her medical history included hypothyroidism, hyperlipidemia, osteopenia, hypertension, and coronary artery disease. She also experienced an inferior myocardial infarction in 1995 requiring angioplasty, surgical repair of her femoral artery in 1995, and hernia repair in 1996. This patient has her INR checked in the anticoagulation clinic and is followed monthly by the clinical pharmacist. Prior to the interaction, her INR was therapeutic for 5 months while she was taking warfarin 1.5 mg/d. The patient admitted to doubling her fish oil dose from 1000 to 2000 mg/d. Without dietary, lifestyle, or medication changes, the INR increased from 2.8 to 4.3 within 1 month. The INR decreased to 1.6 one week after subsequent fish oil reduction, necessitating a return to the original warfarin dosing regimen. DISCUSSION: Fish oil supplementation could have provided additional anticoagulation with warfarin therapy. Fish oil, an omega-3 polyunsaturated fatty acid, consists of eicosapentaenoic acid and docosahexaenoic acid. This fatty acid may affect platelet aggregation and/or vitamin K-dependent coagulation factors. Omega-3 fatty acids may lower thromboxane A(2) supplies within the platelet as well as decrease factor VII levels. Although controversial, this case report illustrates that fish oil can provide additive anticoagulant effects when given with warfarin. CONCLUSIONS: This case reveals a significant rise in INR after the dose of concomitant fish oil was doubled. Patients undergoing anticoagulation therapy with warfarin should be educated about and monitored for possible drug-herb interactions. Pharmacists can play a crucial role in identifying possible drug interactions by asking patients taking warfarin about herbal and other alternative medicine product use. PMID- 14742794 TI - Psoriasiform eruption induced by infliximab. AB - OBJECTIVE: To report a case of psoriasiform eruption induced by infliximab. CASE SUMMARY: A 46-year-old woman with enterocutaneous fistula secondary to Crohn's disease developed pruriginous, erythematous, desquamative plaques on her elbows, knees, hands, and buttocks after receiving the second and third doses of intravenous infliximab. Histologic examination showed a lichenoid pattern. No new cutaneous lesions appeared after cessation of infliximab therapy. DISCUSSION: Several cutaneous reactions secondary to infliximab, a monoclonal antibody against tumor necrosis factor-alfa, have been described. Psoriasiform dermatitis has not been reported as a cutaneous reaction to infliximab, but there have been several previous reports of psoriasiform dermatitis secondary to other drugs. An objective causality assessment revealed that the adverse event was probable. CONCLUSIONS: This is the first report of a clinico-pathologic dissociated pattern of cutaneous reaction showing a histopathologic picture of lichenoid dermatitis resulting from infliximab treatment. PMID- 14742795 TI - Listeria meningitis associated with infliximab. AB - OBJECTIVE: To report a case of Listeria monocytogenes meningitis in a 73-year-old man receiving infliximab for rheumatoid arthritis. CASE SUMMARY: A 73-year-old white man taking infliximab for rheumatoid arthritis developed listeria meningitis following his second dose. He was receiving other immunosuppressants; however, these remained constant immediately prior to the infection. Diagnosis was confirmed with L. monocytogenes isolated in the cerebrospinal fluid. The patient received 21 days of antibiotic therapy and recovered without any complications. DISCUSSION: L. monocytogenes is a gram-positive, non-spore-forming rod that has been associated with the ingestion of undercooked foods. This organism can cause sepsis or meningitis; however, immunocompromised patients, elderly patients, pregnant women, and neonates appear to be at greater risk for this type of infection. Tumor-necrosis factor-alpha (TNF-alpha) plays an important role in resistance to this type of infection, and listeria infections have been reported in 26 patients receiving TNF-alpha inhibitors. In our patient, the listeria infection occurred following his second course of infliximab, which provides a temporal relationship between the listeria infection and infliximab. However, his underlying rheumatoid arthritis and chronic steroid therapy would also increase his risk for a listeria infection. CONCLUSIONS: The listeria infection in our patient was a possible adverse event of infliximab according to the Naranjo probability scale. Because the majority of listeria infections occur in patients who are immunosuppressed, it would be reasonable to provide education for healthcare professionals on preventing these infections in all patients receiving immunosuppressants, including anti-TNF-alpha therapy. Those at risk due to their underlying health conditions should also be monitored closely. PMID- 14742796 TI - Acute delirium associated with combined diphenhydramine and linezolid use. AB - OBJECTIVE: To report a case of delirium with hallucinations presumably caused by the combination of diphenhydramine and linezolid. CASE SUMMARY: A 56-year-old white man was receiving diphenhydramine 300 mg/d for 2 days to treat pruritus caused by a bullous rash possibly induced by vancomycin. He subsequently developed visual and auditory hallucinations, with erratic, aggressive behavior persisting for 3 days. Central anticholinergic syndrome was first suspected, but the long duration and exaggerated response by a patient not prone to anticholinergic toxicity suggest that a second agent may have enhanced the reaction. DISCUSSION: The pharmacodynamic properties of linezolid make this drug a likely contributor to the marked, prolonged effects experienced by this patient. The Naranjo probability scale suggests a possible relationship between the reaction and the combination of diphenhydramine and linezolid. CONCLUSIONS: Drug-induced delirium can occur with several drugs, including diphenhydramine. Linezolid has dopaminergic properties that may enhance the central nervous system effects of anticholinergics. Precautionary monitoring of mental status should be advised when concomitantly administering linezolid with drugs in this class. PMID- 14742797 TI - Ritodrine-induced leukocytoclastic vasculitis in pregnancy. AB - OBJECTIVE: To report 2 cases of pregnant women who presented with leukocytoclastic vasculitis (LV) associated with the administration of ritodrine hydrochloride after undergoing genetic amniocentesis and to review the literature on LV. case summaries: Case 1. A 40-year-old pregnant woman was diagnosed with autoimmune hypothyroidism at week 12 of her third gestation. At week 14, she underwent a genetic amniocentesis and received oral prophylactic treatment with ritodrine for 10 days. At week 16, she presented with fever, epigastric abdominal pain, polyarthritis, microhematuria, and purpura. Skin biopsy showed LV. Case 2. A 34-year-old pregnant woman had developed polyarthralgias and polyarthritis after receiving ritodrine following genetic amniocentesis in her second gestation. She also underwent a genetic amniocentesis in her third gestation and received ritodrine. Five days later, she presented with fever, polyarthritis, and purpura. A skin biopsy demonstrated LV. The rest of her gestation was normal until week 33, when she developed oligohydramnios and a delay of intrauterine fetal growth. A cesarean section was then performed. After birth, the baby developed tachypnea, anemia, splenomegaly, edema, and renal failure; she died on her 15th day of life. DISCUSSION: Ritodrine hydrochloride is a beta(2)-adrenergic agonist that is used in pregnant women as a tocolytic agent. In our 2 patients, there was a time relationship between the administration of ritodrine and the appearance of LV. In both cases, other causes of vasculitis were excluded in a reasonable way. As of November 4, 2003, only one other case has been found in the literature. An objective causality assessment revealed that the reactions to ritodrine were probable and possible, respectively, in our 2 cases. CONCLUSIONS: In pregnant women with autoimmune disease, ritodrine should be used with caution because of its ability to induce vasculitis. PMID- 14742798 TI - Vancomycin ointment for MRSA infection at a cranioplasty site. AB - OBJECTIVE: To report a case illustrating therapeutic success with long-term topical application of aseptic vancomycin ointment to treat methicillin-resistant Staphylococcus aureus (MRSA) infection at a cranioplasty site. CASE SUMMARY: A 63 year-old Japanese woman underwent evacuation of a subdural hematoma complicated by subarachnoid hemorrhage. Subsequent craniotomy for clipping and external decompression of an aneurysm of the neck was followed by cranioplasty using an autologous bone graft. The graft became infected with MRSA, which responded to intravenously infused vancomycin. The graft was then replaced with a ceramic implant. The implant site became reinfected with MRSA. Vancomycin infusion failed on this occasion, despite a favorable in vitro sensitivity test. After obtaining patient consent, investigative treatment was begun using long-term aseptic application of vancomycin 2.5% ointment, resulting in control of the infection and negative cultures. DISCUSSION: The care of an infection at the site of cranioplasty with a ceramic artificial bone implant is difficult. Our patient's infection resolved with the use of vancomycin ointment. In this case, blood concentrations of vancomycin remained below detectable levels, and no adverse effects resulted from application of vancomycin ointment. CONCLUSIONS: Topical administration of vancomycin was more effective than systemic administration in the treatment of our patient's MRSA skull implant infection. No adverse effects from topical treatment were encountered over 3 years. PMID- 14742799 TI - Decreased phenobarbital absorption with charcoal administration for chronic renal failure. AB - OBJECTIVE: To report a case of impaired absorption of orally administered phenobarbital associated with the concomitant administration of activated charcoal, and recovery of the absorption after administration of the 2 drugs was separated by a 1.5-hour interval. CASE SUMMARY: A 78-year-old woman, weighing 50 kg, who had undergone brain surgery was prescribed phenobarbital 120 mg/day for postoperative convulsions. Her serum phenobarbital concentration reached 24.8 microg/mL (therapeutically effective level 10-30). Thereafter, her renal function worsened, and activated charcoal 6 g/day was started. Four months after the start of activated charcoal, blood analysis revealed that the serum phenobarbital concentration was as low as 4.3 microg/mL. The phenobarbital dose was increased to 150 mg/day. Further evaluation revealed that activated charcoal and phenobarbital had been administered concomitantly. The dosage regimen was altered to separate the administration of the agents by at least 1.5 hours. Subsequently, the patient's serum phenobarbital concentration increased to 11.9 microg/mL within 3 weeks. Her serum phenobarbital concentration was measured monthly thereafter and remained stable in the range of 14.8-18.6 microg/mL. DISCUSSION: Our patient's low serum phenobarbital concentration was considered likely to have been due to impaired gastrointestinal absorption of phenobarbital as a result of adsorption of phenobarbital on the activated charcoal. An objective causality assessment showed that the interaction was probable. CONCLUSIONS: Administration of activated charcoal and phenobarbital should be separated by an interval of at least 1.5 hours. PMID- 14742800 TI - Vardenafil treatment for erectile dysfunction. AB - OBJECTIVE: To review the pharmacology, pharmacokinetics, clinical trial data, and adverse effects of vardenafil in the treatment of erectile dysfunction (ED). DATA SOURCES: Literature searches were performed using the MEDLINE database (referenced citations through December 2002), and the references of all identified articles were scanned for additional publications of interest. Unpublished information provided by the manufacturer and proceedings of professional meetings were also evaluated. STUDY SELECTION AND DATA EXTRACTION: All available studies were utilized to obtain information regarding pharmacology. Only human studies were used to gather pharmacokinetic, drug interaction, efficacy, and safety data. DATA SYNTHESIS: Vardenafil is a potent and selective inhibitor of the phosphodiesterase 5 (PDE5) enzyme that has been shown to improve erectile function in several populations of men with ED. Vardenafil has a rapid onset of action, is hepatically metabolized, and has a half-life of 4-6 hours. Clinical trials in otherwise healthy men with ED, men with ED and diabetes, and men with ED and a history of prostatectomy have demonstrated vardenafil's efficacy. Adverse effects appear to be relatively mild in intensity and dose dependent, with 22-61% of subjects reporting adverse effects. CONCLUSIONS: Vardenafil is a safe and effective oral agent for the treatment of ED. Its greater potency and PDE5 selectivity compared with sildenafil appear to confer a lower risk of vision-related adverse effects, but other clinical consequences of these differences are currently unclear. PMID- 14742801 TI - Atomoxetine treatment of attention-deficit/hyperactivity disorder. AB - OBJECTIVE: To review the literature of the first nonstimulant approved for treatment of attention-deficit/hyperactivity disorder (ADHD). DATA SOURCES: Primary literature and review articles were obtained through PubMed/MEDLINE (1966 February 2003). STUDY SELECTION AND DATA EXTRACTION: Literature evaluating atomoxetine in ADHD was selected. Animal data were excluded. DATA SYNTHESIS: Stimulants are currently first-line therapy for ADHD. However, their use raises several concerns including the potential for abuse and adverse effects. Atomoxetine introduces a new therapeutic drug class, selective norepinephrine reuptake inhibitors, for ADHD management. This additional treatment option offers potential advantages over current therapies. CONCLUSIONS: Clinical trials demonstrate that atomoxetine is a safe and effective alternative for ADHD treatment in children and adults; however, its disadvantages may hinder it from becoming a first-line agent. PMID- 14742802 TI - The role of cholesterol and statins in Alzheimer's disease. AB - OBJECTIVE: To briefly discuss the impact of elevated total and low-density lipoprotein cholesterol levels, as well as the potential relationship of hydroxymethylglutaryl coenzyme A reductase inhibitor (statin) use, on the development of Alzheimer's disease (AD). DATA SOURCES: Biomedical literature was accessed through MEDLINE and International Pharmaceutical Abstracts (1966-June 2003). The authors independently reviewed literature for possible inclusion in this article. STUDY SELECTION AND DATA EXTRACTION: Clinical studies were selected and reviewed from the data sources, with special emphasis on those dealing with statin use and AD. DATA SYNTHESIS: The impact of AD is significant, as it is rapidly becoming one of our country's most debilitating and costly diseases. Data from epidemiologic trials indicate that statins may have some protective effect against the development of AD. These trials also allude to theories regarding possible mechanisms of action for this use, data implicating possible superiority of one statin over another, and their lack in certain populations, specifically the very old elderly population. CONCLUSIONS: Preliminary evidence suggests that statins may offer a protective effect against the development of AD. However, review of the literature does not lend credence to the use of statins in the general nondemented population without hyperlipidemia. Potential confounding variables have not been considered in the majority of trials. Placebo-controlled clinical trials are ongoing and should yield more definitive results. PMID- 14742803 TI - Direct thrombin inhibitors for anticoagulation. AB - OBJECTIVE: To review the progress in developing direct thrombin inhibitors (DTIs) for anticoagulation within the context of existing anticoagulation therapies. DATA SOURCES: Searches of MEDLINE (1993-June 2003) were conducted. STUDY SELECTION AND DATA EXTRACTION: We examined English-language articles, human studies, and relevant animal studies, and obtained additional citations from the references of these articles. DATA SYNTHESIS: Because of its pivotal role in hemostasis, thrombin is a key therapeutic target in the treatment and prevention of thromboembolic disorders. Conventional anticoagulant therapies, such as warfarin, unfractionated heparin, and low-molecular-weight heparin, exert their pharmacologic action by indirect thrombin inhibition. Although these agents are effective, each has limitations, prompting a search for more effective, specific, better-tolerated, and convenient anticoagulants. The efficacy and safety of factor Xa inhibitors are being investigated. Furthermore, the development of DTIs such as recombinant hirudin (lepirudin), bivalirudin, and argatroban continues. Challenges in the development of DTIs include establishing a binding affinity for thrombin that is not associated with excessive bleeding, attaining high thrombin specificity, achieving inhibition of both unbound and clot-bound thrombin, and producing an effective, fixed-dose oral anticoagulant to improve the practicality of anticoagulation therapy. Ximelagatran, an oral DTI designed to meet these standards, is currently in Phase III clinical trials. CONCLUSIONS: Significant progress has been made in developing DTIs. The recent emergence of orally administered DTIs may simplify the prevention and treatment of thrombosis. PMID- 14742804 TI - Role of C-reactive protein in cardiovascular disease. AB - OBJECTIVE: To discuss the role of C-reactive protein (CRP) in cardiovascular disease as a predictor of vascular events and identify key factors that increase or decrease this inflammatory marker. DATA SOURCES: Articles were identified through searches of MEDLINE (1966-July 2003), International Pharmaceutical Abstracts (1970-June 2003), and bibliographies of selected articles. Search terms included C-reactive protein, HMG-CoA reductase inhibitors, fenofibrate, niacin, aspirin, estrogen, thiazolidinediones, and raloxifene. data selection and data extraction: All studies relevant to CRP and cardiovascular disease or the effects of pharmacologic and nonpharmacologic interventions on CRP levels were evaluated. All information deemed relevant to this review was included. DATA SYNTHESIS: Numerous studies have shown a strong association between CRP levels and future vascular events (i.e., coronary, cerebrovascular, peripheral vascular disease), with minimal correlation to low-density-lipoprotein cholesterol. Clinical guidelines have recently been published indicating that CRP levels of <1, 1-3, and >3 mg/L correspond to low, moderate, and high risk, respectively, for future vascular events. Drugs including statins, fibrates, niacin, thiazolidinediones, and antiplatelet agents, as well as weight loss and exercise, have demonstrated efficacy in lowering CRP levels. CONCLUSIONS: CRP appears to be a valuable tool for predicting future vascular events in patients striving for primary or secondary prevention of cardiovascular disease. While several pharmacologic and nonpharmacologic interventions have been shown to lower CRP levels, the impact on clinical outcomes requires further study. PMID- 14742805 TI - Point-of-care testing: an introduction. AB - OBJECTIVE: To review available literature and provide perspective on point-of care testing, focusing on the impact it has on treatment outcomes in patient care, the impact it has on the costs of patient care, and the role it has in the delivery of pharmaceutical care. DATA SOURCES: Information was retrieved from MEDLINE English literature searches using PubMed (1965-August 2003) and included search terms of point-of-care testing, near patient testing, pharmaceutical care, pharmacists, outcomes, and economics. Additionally, references from retrieved articles were reviewed to identify literature not detected by literature searches. STUDY SELECTION AND DATA EXTRACTION: Comparative studies, demonstration project reports, and systematic reviews were selected. Other related resources, such as government documents, relevant legislation, and government regulations, were included. Emphasis was placed on comparative studies and demonstration project reports. In the absence of these data, other resources were included. DATA SYNTHESIS: Point-of-care testing devices and technology are increasingly used in the delivery of care and therapeutic decision making. No studies have evaluated the impact of point-of-care testing, by itself, on patient care and outcomes. All studies have incorporated point-of-care testing with changes in the way patient care is delivered and have shown significant improvements when this approach is taken. The cost of point-of-care testing is greater than traditional laboratory testing, but the increased cost may be offset by improvements in the management of patient care, improvements in patient outcomes, and decreased utilization of the healthcare system. Point-of-care testing has been used successfully by pharmacists in disease management programs. Various government regulations and legislation impact the use of point-of-care testing. CONCLUSIONS: Limited data indicate that point-of-care testing, when combined with changes in healthcare delivery systems, may improve patient outcomes and decrease the overall cost of health care. Pharmacists have used point-of-care testing in programs designed to improve patient care but must carefully consider regulations and laws that govern the use of these devices. There is a great need for additional investigation into the use of point-of-care testing in patient care. PMID- 14742806 TI - Monitoring pulmonary function in asthma and COPD: point-of-care testing. AB - OBJECTIVE: To provide pharmacists and other healthcare providers with a better understanding of new monitoring technologies for asthma and chronic obstructive pulmonary disease (COPD) available to clinics and pharmacies. DATA SOURCES: Forty scientific articles were identified through a MEDLINE search (1990-June 2003), additional references listed in articles, and abstracts from scientific meetings. STUDY SELECTION AND DATA EXTRACTION: English-language literature of controlled human clinical studies was reviewed to evaluate the accuracy, reliability, validity, and response of the new monitoring technologies. DATA SYNTHESIS: The In Check DIAL is a pulmonary airflow meter that identifies the most appropriate inhaler for a patient and is useful in determining how efficiently patients use their inhalers. Electronic peak flow meters such as the AirWatch, VMX Wright Mini Log, PiKo-1, and electronic asthma monitor store and download multiple pulmonary function test (PFT) readings to a personal computer, allowing easy identification of patients who are fabricating results. The AirWatch also has the ability to measure forced expiratory volume in 1 second. The Spirophone AG-SP, VM Plus, and Micro DiaryCard spirometer are portable spirometers that can be used at home without the need for supervision. Bronchial challenge tests have been recently standardized and may prove to be beneficial in modifying drug therapy in patients with asthma and COPD. CONCLUSIONS: Despite recent advances in medical technology, monitoring of asthma and COPD has not changed significantly. PFTs continue to be the gold standard for evaluating airway obstruction and/or restriction. Clinical trials that will evaluate outcomes such as decreased number of hospitalizations, emergency department visits, unscheduled visits to physicians, and days absent from school or work are needed to determine the utility of new monitoring technologies. PMID- 14742807 TI - Cost comparisons of olanzapine and risperidone in treating schizophrenia. AB - OBJECTIVE: To review the literature on the healthcare costs associated with olanzapine and risperidone in treating schizophrenia. DATA SOURCES: Published English-language pharmacoeconomic studies on olanzapine and risperidone obtained through a MEDLINE search (1990-May 2003) were selected. Additional studies were identified from a manual search of the references of retrieved articles. data extraction: Based on the identified studies, data were extracted on various treatment costs associated with the use of antipsychotic drugs, concomitant drugs from other classes, inpatient care, outpatient care, and emergency care. Emphasis was placed on studies directly comparing olanzapine and risperidone. DATA SYNTHESIS: Both olanzapine and risperidone were generally associated with a trend of decrease in total medical costs compared with typical antipsychotics. When directly comparing the drugs, some studies found significant cost savings in favor of olanzapine and some suggested risperidone to save total costs. Still others showed no significant difference in total costs between the 2 drug regimens. CONCLUSIONS: While both olanzapine and risperidone appear to be more cost saving than typical antipsychotics, the literature offers no conclusive evidence to determine the comparative advantage of one versus another in terms of total cost outcomes. Major factors that contribute to the inconclusive findings may include across-study variations in populations, design, outcome measures, dosage, severity of illness, inclusion criteria, and statistical methodologies. PMID- 14742808 TI - Use of beta-blockers in patients with COPD. AB - OBJECTIVE: To evaluate the safety and cardiovascular benefits of beta-blocker therapy in patients with chronic obstructive pulmonary disease (COPD). DATA SOURCES: Clinical literature was accessed through MEDLINE (1966-February 2003). Key search terms included chronic obstructive pulmonary disease and adrenergic beta-antagonists. DATA SYNTHESIS: beta-Blockers are often avoided in patients with COPD because of fear of bronchoconstriction, despite the known cardiovascular mortality benefits. A review of studies evaluating the use of beta blockers in COPD was undertaken. CONCLUSIONS: The literature supports the safety and mortality benefits of using beta-blockers in COPD. Patients with mild to moderate COPD should receive cardioselective beta-blocker therapy when a strong indication exists. PMID- 14742809 TI - Antipyretic efficacy and safety of ibuprofen and acetaminophen in children. AB - OBJECTIVE: To evaluate the antipyretic effects and safety of ibuprofen compared with acetaminophen in febrile children. DATA SOURCES: Searches of MEDLINE (1966 November 2003) and EMBASE (1988-November 2003) were conducted using the terms ibuprofen and acetaminophen. Bibliographies of selected articles were reviewed. DATA SYNTHESIS: Ibuprofen was significantly more effective than acetaminophen in reducing fever after a single dose. Ibuprofen was found to be more effective after 6 hours, but not after a longer period of time. Studies with multiple doses have also failed to show that one drug is better than the other. CONCLUSIONS: The efficacy and effectiveness of acetaminophen and ibuprofen in their recommended dosages are similar, with slightly more beneficial effects shown with ibuprofen. PMID- 14742810 TI - Recent changes in quality in Japanese clinical trials. AB - BACKGROUND: Japanese clinical trials have some unique characteristics in both design and conduct. We have studied recent changes in quality in Japanese clinical trials. OBJECTIVE: To describe quantitatively recent drastic changes in Japanese clinical trial environments by comparing the results of Good Clinical Practice (GCP) audits conducted from April 1997 to March 2000 (fiscal year [FY] 1997-1999) with those from April 2000 to March 2001 (FY2000). METHODS: The numbers and proportions of various types of deficiencies described in GCP audit reports were compared between the 2 periods. The audit findings in the former period were based on official audits that covered 331 hospitals and 775 trials. The audits in the latter period targeted 123 hospitals and 279 trials. In both periods, inspections were undertaken by the Organization for Pharmaceutical Safety and Research (OPSR). RESULTS: The total number of deficiencies detected in GCP audits in the former 3-year period (FY1997-1999) was 1529; the number in the next single year (FY2000) was 1151. The total number of deficiencies detected and reported was more than double on an annual basis between the periods. By category of deficiencies, the proportion of protocol deviations increased from 14.7% (n = 225) to 41.4% (n = 477), while the proportion of errors in Case Report Forms decreased from 43.6% (n = 666) to 34.1% (n = 392). CONCLUSIONS: This study shows that the protocol deviations increased in FY2000. The increase in deficiencies may be associated with the regulatory change of applicable standards, increasing attention of the OPSR to such deficiencies, difficulties in improving investigators' behaviors during a short period of time, and insufficient numbers of support staff including clinical research coordinators in research institutions. PMID- 14742811 TI - A new collaboration enhances The Annals Online. PMID- 14742812 TI - What is optimal for antibiotic studies in acute otitis media? PMID- 14742813 TI - Adherence or compliance? Changes in terminology. PMID- 14742814 TI - Challenges to the FDA approval of mifepristone. PMID- 14742815 TI - Therapeutic and economic consequences of OTC loratadine. PMID- 14742816 TI - Myelodysplastic syndrome associated with low-dose methotrexate in rheumatoid arthritis. PMID- 14742817 TI - Aseptic meningitis possibly associated with celecoxib. PMID- 14742818 TI - Cutaneous ulceration induced by interferon alfa. PMID- 14742819 TI - Prediction of milk/plasma concentration ratio of drugs. PMID- 14742820 TI - Prediction of milk/plasma concentration ratio of drugs. PMID- 14742821 TI - Retraction: acute myelogenous leukemia following mitoxantrone treatment for multiple sclerosis. PMID- 14742826 TI - Osteoporosis: point-of-care testing. AB - OBJECTIVE: To review the literature concerning the utility of point-of-care (POC) testing devices for the diagnosis and management of osteoporosis. DATA SOURCES: Articles were identified from a MEDLINE search (1993-June 2003). Additional references were obtained from cross-referencing the bibliographies of selected articles. STUDY SELECTION AND DATA EXTRACTION: After evaluation of clinical trials and select review articles, articles comparing peripheral dual-energy absorptiometry (pDXA) or quantitative ultrasound (QUS) with central DXA (cDXA) measurements were emphasized in this analysis. DATA SYNTHESIS: Sensitivity for detecting osteoporosis by QUS or pDXA varies widely (range 35-75%). Using adjusted T-score cutoffs increases sensitivity to 85-95%, at the price of reducing device specificity to 23-49%. Many states require a radiology technician to perform pDXA tests. CONCLUSIONS: POC testing with peripheral devices should only be considered in areas with limited access to cDXA or for women who initially refuse cDXA testing. T scores of-1.0 or less with POC testing typically require further evaluation via cDXA. Many states require pDXAs to be performed by certified radiology technologists, making QUS use more feasible for pharmacists. POC testing should not be used for assessing response to osteoporosis therapy. PMID- 14742827 TI - Ephedra-associated cardiomyopathy. AB - OBJECTIVE: To report 2 cases of cardiomyopathy associated with use of dietary supplements containing ephedra. case summaries: A 19-year-old white man presented to the emergency department (ED) complaining of exertional shortness of breath and episodic chest pain radiating to the left arm. Left heart catheterization revealed no significant coronary artery disease, a dilated left ventricle, and global hypokinesis. He was discharged home 5 days after admission on standard therapies for heart failure, but died 5 weeks later. A 21-year-old white man presented to the ED with recurrent chest pain and was diagnosed with myopericarditis. An echocardiogram showed global hypokinesis with an ejection fraction of 40-50%. He was treated for myopericarditis with standard therapies for heart failure. An objective causality assessment probability scale revealed that an adverse drug reaction was possible between cardiomyopathy and ephedra use in these 2 patients. Both of these cases have been reported to MedWatch. DISCUSSION: Ephedrine is a potent sympathomimetic agent with direct and indirect effects on adrenergic receptors to cause increases in heart rate, blood pressure, cardiac output, and vascular resistance. The adverse effects of adrenergic stimulation are well known in cardiomyopathy, inducing direct and indirect myocyte toxicity. CONCLUSIONS: It is well documented that ephedra, through its sympathomimetic effects, can cause a range of cardiovascular toxicities including myocarditis, arrhythmias, myocardial infarction, cardiac arrest, and sudden death. PMID- 14742828 TI - Heparin-induced hyponatremia. AB - OBJECTIVE: To report a case of hyponatremia in a patient receiving systemic unfractionated heparin (UFH) therapy and parenteral nutrition. CASE SUMMARY: A 70 year-old African American woman was started on parenteral nutrition for postoperative ileus following an elective surgical procedure. Three days later, she was diagnosed with a pulmonary embolism and intravenous UFH therapy was initiated. During the 7-day course of UFH therapy, the patient's serum sodium concentration steadily declined and urine sodium concentration progressively increased. Physical examination revealed no signs or symptoms of hypo- or hypervolemia. The patient's serum potassium concentration increased modestly, although significant hyperkalemia was not observed. After discontinuation of UFH, serum concentrations of both sodium and potassium returned to baseline levels. DISCUSSION: Although heparin-induced hyperkalemia is well documented, cases associated with substantial hyponatremia have been reported less frequently. An objective causality assessment revealed that the adverse drug reaction was probable in this case. Hyponatremia and hyperkalemia result from the antagonism of aldosterone by UFH within the zona glomerulosa of the adrenal glands. CONCLUSIONS: The use of UFH may result in significant hyponatremia as well as hyperkalemia. Reversal of these electrolyte disturbances occurs after discontinuation of heparin. PMID- 14742829 TI - Economic consequences of venous thromboembolism following major orthopedic surgery. AB - BACKGROUND: Venous thromboembolism (VTE) is a frequent and potentially costly complication of major orthopedic surgery. OBJECTIVE: To estimate the economic consequences of VTE following major orthopedic surgery. METHODS: Using a large healthcare claims database, we identified all patients who underwent total hip replacement, major knee surgery, or hip fracture repair from January 1993 to December 1998. Patients with clinical VTE (cases) were identified based on a diagnosis of deep vein thrombosis or pulmonary embolism within 90 days of surgery (index admission) and > or =1 prescription for warfarin or unfractionated heparin within 30 days of the date of initial VTE diagnosis. Each case was matched (using age and procedure type) to 2 randomly selected patients who did not have any claims for clinical VTE (matched controls). Utilization and billed charges were then examined over a 90-day period following admission. Cases were stratified based on whether VTE was first noted during the index admission or thereafter. RESULTS: A total of 11 960 patients were identified who underwent total hip replacement, major knee surgery, or hip fracture repair (n = 3171, 3955, 4834, respectively). Over a 90-day period, 259 patients (2.2%) developed clinical VTE. Most cases (61.8%) occurred after hospital discharge. For patients with in hospital VTE, mean length of stay for the index admission was 4.5 days longer than that of matched controls (11.1 vs 6.6); by day 90, there was a 5.4-day difference in total hospital days. Mean billed charges for the index admission were 17,552 higher US dollars (52,037 US dollars vs 34,485 US dollars); the difference rose to 18,834 US dollars by day 90 (54,480 US dollars vs 35,646 US dollars). For patients who developed clinical VTE following hospital discharge, there was a 3.4-day difference in total hospital days at day 90 (10.2 vs 6.8) as a result of readmissions for VTE; mean total billed charges at day 90 were 5765 US dollars higher (41,411 US dollars vs 35,646 US dollars). CONCLUSIONS: Among patients who have undergone major orthopedic surgery, the economic consequences of VTE are substantial, regardless of the setting in which it occurs. PMID- 14742830 TI - Comment: Reasons for early abacavir discontinuation in HIV-infected patients. PMID- 14742831 TI - Therapy switching in patients receiving long-acting opioids. AB - BACKGROUND: Patterns of therapy switching in patients receiving long-acting opioids have not been well documented. OBJECTIVE: To compare therapy switching among patients beginning treatment with controlled-release (CR) oxycodone, transdermal fentanyl, or CR morphine sulfate. METHODS: Using a US healthcare claims database, we identified patients beginning treatment with CR oxycodone, transdermal fentanyl, or CR morphine sulfate between July 1, 1998, and December 31, 1999. We compiled claims for each patient for 6 months following therapy initiation and compared the incidence of therapy switching among the 3 groups. We also estimated total healthcare charges for patients who switched therapy versus those who did not. RESULTS: We identified 1931, 668, and 449 patients beginning therapy with CR oxycodone, transdermal fentanyl, and CR morphine sulfate, respectively; 16.7%, 25.0%, and 35.9%, respectively, had cancer. For patients without cancer, rates of therapy switching at 6 months were 10.6% (CR oxycodone), 19.0% (transdermal fentanyl), and 26.0% (CR morphine sulfate); for those with cancer, rates were 23.8%, 24.6%, and 29.8%, respectively. Multivariate hazard ratios (vs CR morphine sulfate) for therapy switching in patients without cancer were 0.36 (95% CI, 0.27 to 0.47) for CR oxycodone and 0.69 (0.51 to 0.94) for transdermal fentanyl; for those with cancer, corresponding hazard ratios were 0.72 (0.50 to 1.03) and 0.76 (0.50 to 1.16). Total healthcare charges were significantly (p < 0.01) higher for patients who switched therapy than those who did not (23,965 US dollars vs 14,299 US dollars in pts. without cancer; 58,259 US dollars vs 39,618 US dollars for those with cancer). CONCLUSIONS: Patients without cancer who receive CR oxycodone or transdermal fentanyl are less likely to switch therapy than those receiving CR morphine sulfate. Total healthcare charges are higher for patients who switch therapy. PMID- 14742832 TI - Acetylcysteine treatment for non-acetaminophen-induced acute liver failure. AB - OBJECTIVE: To evaluate the effectiveness of intravenous acetylcysteine in the treatment of non-acetaminophen-induced acute liver failure (ALF). DATA SOURCES: A search of MEDLINE (1966-March 2003), International Pharmaceutical Abstracts (1970 2003), and Cochrane Library (2003, issue 3) databases was conducted, using the search terms acetylcysteine, non-acetaminophen-induced hepatic failure, liver failure, intravenous, and treatment. DATA SYNTHESIS: All of the studies found were small and do not provide conclusive evidence that acetylcysteine benefits this subgroup of patients. Microvascular regional benefits were seen, but clinical outcomes have not been studied. CONCLUSIONS: Intravenous acetylcysteine should not be used routinely for treatment of non-acetaminophen-induced ALF. Further large-scale studies are needed to evaluate clinical outcomes. PMID- 14742833 TI - Recurrent ibuprofen-induced aseptic meningitis. AB - OBJECTIVE: To report a case of recurrent aseptic meningitis temporally associated with the use of ibuprofen. CASE SUMMARY: A previously well 51-year-old white man presented with acute confusion and aphasia 7 days after taking a variety of nonprescription medications, including ibuprofen. Imaging of the brain was unremarkable, and lumbar puncture revealed lymphocytic pleocytosis with an elevated protein level. The symptoms improved shortly after admission, and no infectious cause was identified. Two weeks later, the patient was readmitted with similar symptoms beginning immediately after resumption of ibuprofen. His symptoms resolved promptly after ibuprofen was discontinued. DISCUSSION: Drug induced aseptic meningitis is an unusual complication of drug therapy. Nonsteroidal antiinflammatory drugs (NSAIDs), particularly ibuprofen, are among the most commonly implicated agents, but rechallenge with the suspected agent is uncommon. Use of an objective causality tool indicated a probable relationship between ibuprofen and development of aseptic meningitis in our patient. CONCLUSIONS: Clinicians should consider NSAIDs as potential causes of aseptic meningitis, especially in patients with recurrent illness and no obvious infectious cause. A detailed drug history is invaluable in the assessment of such patients, with particular attention to nonprescription medications such as ibuprofen. PMID- 14742834 TI - Stability of lisinopril in two liquid dosage forms. AB - BACKGROUND: Lisinopril is used in pediatric patients with hypertension. It is not commercially available as a liquid. Little is known about the stability of lisinopril in extemporaneously prepared liquid dosage forms. OBJECTIVE: To determine the stability of lisinopril in 2 oral suspensions stored at 4 and 25 degrees C in plastic prescription bottles. METHODS: Five bottles contained methylcellulose 1%:simple syrup NF (1:13) and the other 5 bottles had Ora Plus Ora Sweet (1:1) at a lisinopril concentration of 1 mg/mL. Three samples were collected from each bottle at 0, 7, 14, 28, 42, 56, 70, and 91 days and analyzed by stability-indicating HPLC analytical method (n = 15). RESULTS: At 4 degrees C, the mean +/- SD concentration of lisinopril remained >95.1 +/- 1.8% of the initial concentration in the methylcellulose formulation and 95.1 +/- 3.2% of the initial concentration in the Ora Plus-Ora Sweet formulation throughout the 91-day study period. At 25 degrees C, the mean concentration of lisinopril remained >92.4 +/- 2.2% of the initial concentration in the methylcellulose formulation for 8 weeks and 95.8 +/- 2.3% of the initial concentration in the Ora Plus-Ora Sweet formulation throughout the 91-day study period. No changes in physical appearance in any samples were seen during this period. CONCLUSIONS: Lisinopril can be prepared in either of 2 liquid dosage forms and stored for at least 13 weeks under refrigeration and 8 weeks at room temperature. PMID- 14742835 TI - 11beta-hydroxysteroid dehydrogenase type 1 in obesity. PMID- 14742836 TI - Lack of association between human G-protein beta3 subunit variant and overweight in Japanese workers. AB - We examined the association of the G-protein beta3 subunit gene (GNB3) C825T polymorphism with overweight in Japanese workers. This cross-sectional study used multivariate analysis to investigate whether a polymorphism in the C825T polymorphism was associated independently with overweight when factors such as age and lifestyle were taken into account. The study in 1453 men and 1172 women involved identifying subjects with the C825T genotype using the polymerase chain reaction, followed by restriction fragment-length polymorphism analysis. Overweight was defined as a BMI > or = kg/m(2). Genotype distributions for C825T in overweight men (CC = 80, CT = 162, TT = 80) and women (CC = 52, CT = 91, TT = 40) were not significantly different from normal-weight men (CC = 278, CT = 588, TT = 265) and women (CC = 242, CT = 549, TT = 198). The allele distributions were also not significantly different between either sex. The power of the study was estimated as 98% in men and 81% in women based on the allelic frequencies reported in a previous positive study in Chinese subjects. Multiple logistic regression analysis showed that the genotype was not significantly associated with overweight. In conclusion, this study indicated that the GNB3 C825T polymorphism is not a significant factor for overweight in Japanese people. PMID- 14742837 TI - Regulation of 11beta-HSD genes in human adipose tissue: influence of central obesity and weight loss. AB - OBJECTIVES: The activity of adipose 11beta-hydroxysteroid dehydrogenase (11beta HSD) 1 is increased in obese subjects, and animal data suggest that increased cortisol formation in adipose tissue contributes to the development of the metabolic syndrome. The aim of this study was to determine whether up-regulation of human adipose 11beta-HSD1 in obesity can also be found at the gene expression level. RESEARCH METHODS AND PROCEDURES: 11beta-HSD gene expression in subcutaneous adipose tissue biopsies of 70 postmenopausal women was studied by real-time reverse-transcription polymerase chain reaction. The influence of weight reduction and in vitro effects of several modulators of adipocyte gene expression on 11beta-HSD genes in human adipocytes were also studied. RESULTS: The 11beta-HSD1 gene was highly expressed in human adipose tissue. 11beta-HSD2 mRNA was also detectable at lower levels. Adipose 11beta-HSD1 gene expression was increased by two-fold and was positively correlated with waist circumference and homeostasis model assessment index of insulin resistance. 11beta-HSD2 gene expression was reduced by half in obese women. Weight reduction did not change gene expression levels of 11beta-HSD1 or 11beta-HSD2. Cortisol increased 11beta HSD1 gene expression in isolated human adipocytes in vitro, whereas estradiol, triiodothyronine, angiotensin II, and pioglitazone had no influence. DISCUSSION: Our data suggest that increased expression of the 11beta-HSD1 gene is associated with metabolic abnormalities in obese women and that increased expression of this gene may contribute to the previously reported increased local conversion of cortisone to cortisol in adipose tissue of obese individuals. PMID- 14742838 TI - State-level estimates of annual medical expenditures attributable to obesity. AB - OBJECTIVE: To provide state-level estimates of total, Medicare, and Medicaid obesity-attributable medical expenditures. RESEARCH METHODS AND PROCEDURES: We developed an econometric model that predicts medical expenditures. We used this model and state-representative data to quantify obesity-attributable medical expenditures. RESULTS: Annual U.S. obesity-attributable medical expenditures are estimated at $75 billion in 2003 dollars, and approximately one-half of these expenditures are financed by Medicare and Medicaid. State-level estimates range from $87 million (Wyoming) to $7.7 billion (California). Obesity-attributable Medicare estimates range from $15 million (Wyoming) to $1.7 billion (California), and Medicaid estimates range from $23 million (Wyoming) to $3.5 billion (New York). DISCUSSION: These estimates of obesity-attributable medical expenditures present the best available information concerning the economic impact of obesity at the state level. Policy makers should consider these estimates, along with other factors, in determining how best to allocate scarce public health resources. However, because they are associated with large SE, these estimates should not be used to make comparisons across states or among payers within states. PMID- 14742839 TI - Insulin action on expression of novel adipose genes in healthy and type 2 diabetic subjects. AB - OBJECTIVE: Adipose tissue secretes several molecules that may participate in metabolic cross-talk to other insulin-sensitive tissues. Thus, adipose tissue is a key endocrine organ that regulates insulin sensitivity in other peripheral insulin target tissues. We have studied the expression and acute insulin regulation of novel genes expressed in adipose tissue that are implicated in the control of whole body insulin sensitivity. RESEARCH METHODS AND PROCEDURES: Expression of adiponectin, c-Cbl-associated protein (CAP), 11-beta hydroxysteroid dehydrogenase type 1 (11beta-HSD-1), and sterol regulatory element binding protein (SREBP)-1c was determined in subcutaneous adipose tissue from type 2 diabetic and age- and BMI-matched healthy men by real-time polymerase chain reaction analysis. RESULTS: Expression of adiponectin, CAP, 11beta-HSD-1, and SREBP-1c was similar between healthy and type 2 diabetic subjects. Insulin infusion for 3 hours did not affect expression of CAP, 11beta-HSD-1, or adiponectin mRNA in either group. However, insulin infusion increased SREBP-1c expression by 80% in healthy, but not in type 2 diabetic, subjects. DISCUSSION: Our results provide evidence that insulin action on SREBP-1c is dysregulated in adipose tissue from type 2 diabetic subjects. Impaired insulin regulation on gene expression of select targets in adipose tissue may contribute to the pathogenesis of type 2 diabetes. PMID- 14742840 TI - Regulation of lipolysis and lipoprotein lipase after weight loss in obese, postmenopausal women. AB - OBJECTIVE: To test the hypothesis that the greater beta-adrenoceptor (beta-AR) stimulated lipolysis and sensitivity (half-maximal lipolytic response) in abdominal (ABD) adipocytes, greater gluteal (GLT) adipose tissue-lipoprotein lipase (AT-LPL) activity, and dyslipidemia associated with obesity in older women are modifiable by weight loss (WL) and are not due to menopause or aging. RESEARCH METHODS AND PROCEDURES: The metabolic effects of 6 months of hypocaloric diet and low-intensity walking WL program on the regional regulation of in vitro lipolysis and AT-LPL activity in subcutaneous ABD and GLT adipocytes were measured in 34 obese (48.7 +/- 0.7% body fat, mean +/- SE) postmenopausal (59 +/- 1 years) white women. RESULTS: The lipolytic responsiveness to the beta-AR agonist isoproterenol and basal lipolysis in the presence of 1 U/mL adenosine deaminase-uninhibited (lipolysis) were greater (p < 0.01) in ABD than GLT adipocytes before and after WL, but there were no regional differences in postreceptor (dibutyryl 3',5'-cyclic adenosine monophosphate)-stimulated lipolysis. beta-AR sensitivity was greater in ABD than GLT adipocytes before (p < 0.01) but not after WL. Regional AT-LPL did not change after WL, but the change in the activity of ABD (but not GLT) AT-LPL correlated with the baseline adenosine deaminase-uninhibited lipolysis (r = 0.38, p = 0.03). There were no relationships between the declines in plasma triglyceride or increases in high density lipoprotein cholesterol associated with WL and the changes in regional fat cell metabolism. DISCUSSION: Thus, despite improving lipoprotein lipid profiles in obese, postmenopausal women, WL does not affect the regulation of regional fat metabolism, and a greater tonic inhibition of basal lipolysis by endogenous adenosine may increase the activity of AT-LPL after WL and predispose older women to develop ABD adiposity. PMID- 14742841 TI - Bone and gastric bypass surgery: effects of dietary calcium and vitamin D. AB - OBJECTIVE: To examine bone mass and metabolism in women who had previously undergone Roux-en-Y gastric bypass (RYGB) and determine the effect of supplementation with calcium (Ca) and vitamin D. RESEARCH METHODS AND PROCEDURES: Bone mineral density and bone mineral content (BMC) were examined in 44 RYGB women (> or = 3 years post-surgery; 31% weight loss; BMI, 34 kg/m(2)) and compared with age- and weight-matched control (CNT) women (n = 65). In a separate analysis, RYGB women who presented with low bone mass (n = 13) were supplemented to a total 1.2 g Ca/d and 8 microg vitamin D/d over 6 months and compared with an unsupplemented CNT group (n = 13). Bone mass and turnover and serum parathyroid hormone (PTH) and 25-hydroxyvitamin D were measured. RESULTS: Bone mass did not differ between premenopausal RYGB and CNT women (42 +/- 5 years), whereas postmenopausal RYGB women (55 +/- 7 years) had higher bone mineral density and BMC at the lumbar spine and lower BMC at the femoral neck. Before and after dietary supplementation, bone mass was similar, and serum PTH and markers of bone resorption were higher (p < 0.001) in RYGB compared with CNT women and did not change significantly after supplementation. DISCUSSION: Postmenopausal RYGB women show evidence of secondary hyperparathyroidism, elevated bone resorption, and patterns of bone loss (reduced femoral neck and higher lumbar spine) similar to other subjects with hyperparathyroidism. Although a modest increase in Ca or vitamin D does not suppress PTH or bone resorption, it is possible that greater dietary supplementation may be beneficial. PMID- 14742842 TI - Performance of two self-report measures for evaluating obesity and weight loss. AB - OBJECTIVES: To evaluate performance of the Obesity and Weight-Loss Quality-of Life (OWLQOL) and Weight-Related Symptoms (WRSM) measures. RESEARCH METHODS AND PROCEDURES: Four studies of obese persons 18 to 75 years of age were analyzed: a 12-week initial validation study, a clinical trial using blinded endpoint data at 50 to 83 weeks, and community studies conducted in the U.S. and Europe. Fifty-six initial validation study subjects visited 1 week after screening to evaluate reproducibility. RESULTS: Overall, 6107 obese persons completed one assessment, 291 completed follow-up at 12 weeks, and 642 at >50 weeks. Psychometric analyses resulted in a 17-item OWLQOL with a single score tested on five samples that was internally consistent (alpha values > 0.90) and reproducible (intraclass correlation coefficient > 0.95). The OWLQOL score (higher is better) was associated, as expected, with the symptom measure (lower is better, -0.54), generic quality of life measure (0.53), and measures of physical (0.40) and mental functioning (0.47). The 20-item WRSM was internally consistent (alpha = 0.87) and reproducible (intraclass correlation coefficient = 0.83). The OWLQOL discriminated between genders (p < 0.001), presence of disability days (p < 0.05), levels of BMI (p < 0.05), and levels of symptom bothersomeness (p < 0.001). Evaluation at 12 weeks yielded an effect size for > or = 2.5% weight loss of 0.77 for the OWLQOL and -0.54 for the WRSM. At > or = 50 weeks for > or = 10% weight change, effect sizes were 1.63 and -0.73, respectively. DISCUSSION: The OWLQOL and WRSM are brief, valid, reproducible, and responsive self-reported outcomes for evaluating obesity and weight loss. PMID- 14742843 TI - Childhood overweight and academic performance: national study of kindergartners and first-graders. AB - OBJECTIVE: To examine the association between children's overweight status in kindergarten and their academic achievement in kindergarten and first grade. RESEARCH METHODS AND PROCEDURES: The data analyzed consisted of 11,192 first time kindergartners from the Early Childhood Longitudinal Study, a nationally representative sample of kindergartners in the U.S. in 1998. Multivariate regression techniques were used to estimate the independent association of overweight status with children's math and reading standardized test scores in kindergarten and grade 1. We controlled for socioeconomic status, parent-child interaction, birth weight, physical activity, and television watching. RESULTS: Overweight children had significantly lower math and reading test scores compared with non-overweight children in kindergarten. Both groups were gaining similarly on math and reading test scores, resulting in significantly lower test scores among overweight children at the end of grade 1. However, these differences, except for boys' math scores at baseline (difference = 1.22 points, p = 0.001), became insignificant after including socioeconomic and behavioral variables, indicating that overweight is a marker but not a causal factor. Race/ethnicity and mother's education were stronger predictors of test score gains or levels than overweight status. DISCUSSION: Significant differences in test scores by overweight status at the beginning of kindergarten and the end of grade 1 can be explained by other individual characteristics, including parental education and the home environment. However, overweight is more easily observable by other students compared with socioeconomic characteristics, and its significant (unadjusted) association with worse academic performance can contribute to the stigma of overweight as early as the first years of elementary school. PMID- 14742844 TI - BMI and health-related quality of life in adults 65 years and older. AB - OBJECTIVE: To examine relationships of BMI with health-related quality of life in adults 65 years and older. RESEARCH METHODS AND PROCEDURES: In 1996, a health survey was mailed to all surviving participants > or = 65 years old from the Chicago Heart Association Detection Project in Industry Study (1967 to 1973). The response rate was 60%, and the sample included 3981 male and 3099 female respondents. BMI (kilograms per meter squared) was classified into four groups: underweight (<18.5), normal weight (18.5 to 24.9), overweight (25.0 to 29.9), and obese (> or = 30.0). Main outcome measures were Health Status Questionnaire-12 scores (ranging from 0 to 100) assessing eight domains: health perception, physical functioning, role limitations-physical, bodily pain, energy/fatigue, social functioning, role limitations-mental, and mental health. The higher the score, the better the outcome. RESULTS: With adjustment for age, race, education, smoking, and alcohol intake, obesity was associated with lower health perception and poorer physical and social functioning (women only) but not impaired mental health. Overweight was associated with impaired physical well-being among women only. Both underweight men and women reported impairment in physical, social, and mental well-being. For example, multivariable-adjusted health perception domain scores for women were 50.8 (underweight), 62.7 (normal weight), 60.5 (overweight), and 52.1 (obese), respectively. Associations weakened but remained significant with further adjustment for comorbidities. DISCUSSION: Compared with normal-weight people, both underweight and obese older adults reported impaired quality of life, particularly worse physical functioning and physical well-being. These results reinforce the importance of normal body weight in older age. PMID- 14742845 TI - Insulin sensitivity, cardiorespiratory fitness, and physical activity in overweight Hispanic youth. AB - OBJECTIVE: To determine whether cardiorespiratory fitness and/or physical activity (PA) were related to measures of insulin sensitivity and secretion independent of body composition in overweight Hispanic children. RESEARCH METHODS AND PROCEDURES: Ninety-five Hispanic children (n = 55 boys; n = 40 girls; 8 to 13 years old) participated in this investigation. The frequently sampled intravenous glucose tolerance test was used to determine the insulin sensitivity index (SI), the acute insulin response, and the disposition index. Cardiorespiratory fitness [maximal oxygen uptake (VO(2max))] was evaluated using a treadmill protocol, and PA was determined by an interviewer-administered questionnaire. Body composition was measured using DXA. RESULTS: Unadjusted correlations indicated that VO(2max) (milliliters of O(2) per minute) was negatively related to SI (r = -0.46, p < 0.05) and disposition index (r = -0.31, p < 0.05) and positively associated with fasting insulin (r = 0.29, p < 0.05), but these relationships were no longer significant once gender, Tanner stage, fat mass, and soft lean tissue mass were included as covariates (all p > 0.05). Multivariate linear regression analysis showed that body fat mass explained 53% of the variance in SI and that VO(2max) (milliliters of O(2) per minute) was not independently related to SI. Cardiorespiratory fitness was positively related to both fat mass (r = 0.43, p < 0.001) and soft lean tissue mass (r = 0.89, p < 0.001). PA was not related to any measure of insulin sensitivity and secretion. DISCUSSION: Cardiorespiratory fitness, as determined by VO(2max) (milliliters of O(2) per minute), was not independently related to insulin sensitivity or secretion, suggesting that VO(2max) influences insulin dynamics indirectly through fat mass. PMID- 14742847 TI - Computerized determination of adipocyte size. AB - OBJECTIVE: Fat cell size is a fundamental parameter in the study of adipose tissue metabolism, because it markedly influences the cellular rates of metabolism. Previous techniques for the sizing of adipocytes are often complicated or time-consuming. The aim of this study was to develop a new, computerized method for rapid and accurate determination of adipocyte size in a cell suspension obtained by incubating human or rat adipose tissue biopsies with collagenase. RESEARCH METHODS AND PROCEDURES: The cell suspension was placed between a siliconized glass slide and a cover slip. Using the reference method [designated as (R)], the cell diameters were determined manually using a microscope with a calibrated ocular. The new method presented here [designated as (C)] was based on computerized image analysis. RESULTS: After two well-defined corrective adjustments, measurements with (R) and (C) agreed very well. The small remaining differences seemed, in fact, to depend on inconsistencies in (R). DISCUSSION: We propose that (C) constitutes a valuable tool to study fat cell size, because this method is fast and allows the assessment of a sufficient number of cells to get reliable data on size distribution. Furthermore, images of cell preparations may be stored for future reference. PMID- 14742846 TI - Predictors of fat stereotypes among 9-year-old girls and their parents. AB - OBJECTIVE: To assess familial links in fat stereotypes and predictors of stereotypes among girls and their parents. RESEARCH METHODS AND PROCEDURES: Fat stereotypes were assessed using a questionnaire developed for this study. Participants indicated the extent to which they agreed with nine statements about thin people (e.g., thin people are smart) and the same statements about fat people (e.g., fat people are smart). Predictors of fat stereotypes that were examined include weight status (BMI; girls and parents), education (parents), income (parents), self-investment in physical appearance (parents), maladaptive eating attitudes (girls), and parenting practices and peer interactions focused on body shape and weight loss (girls). RESULTS: Girls and parents exhibited fat stereotypes. Fathers who were more educated and had a higher family income were more likely to endorse fat stereotypes, as were mothers and fathers with a high investment in their physical appearance. Although no associations were found between girls' and parents' fat stereotypes, girls were more likely to endorse fat stereotypes when interactions with parents and peers focused on body shape and weight loss. Girls were also more likely to endorse fat stereotypes when they reported higher levels of maladaptive eating attitudes. No associations were found between weight status and fat stereotypes. DISCUSSION: Although there was no association between girls' and parents' fat stereotypes, girls were more likely to express negative attitudes about obesity and obese persons when parenting practices and interactions with peers promoted a lean body type, suggesting that there may be an implicit link between the lean ideal and fat stereotypes. PMID- 14742848 TI - The role of overweight and obesity in calcium oxalate stone formation. AB - OBJECTIVE: The aim of the study was to assess the influence of overweight and obesity on the risk of calcium oxalate stone formation. RESEARCH METHODS AND PROCEDURES: BMI, 24-hour urine, and serum parameters were evaluated in idiopathic calcium oxalate stone formers (363 men and 164 women) without medical or dietetic pretreatment. RESULTS: Overweight and obesity were present in 59.2% of the men and in 43.9% of the women in the study population. Multiple linear regression analysis revealed a significant positive relationship between BMI and urinary uric acid, sodium, ammonium, and phosphate excretion and an inverse correlation between BMI and urinary pH in both men and women, whereas BMI was associated with urinary oxalate excretion only among women and with urinary calcium excretion only among men. Serum uric acid and creatinine concentrations were correlated with BMI in both genders. Because no association was established between BMI and urinary volume, magnesium, and citrate excretion, inhibitors of calcium oxalate stone formation, the risk of stone formation increased significantly with increasing BMI among both men and women with urolithiasis (p = 0.015). The risk of calcium oxalate stone formation, median number of stone episodes, and frequency of diet-related diseases were highest in overweight and obese men. DISCUSSION: Overweight and obesity are strongly associated with an elevated risk of stone formation in both genders due to an increased urinary excretion of promoters but not inhibitors of calcium oxalate stone formation. Overweight and obese men are more prone to stone formation than overweight women. PMID- 14742849 TI - Upper and lower body adipose tissue function: a direct comparison of fat mobilization in humans. AB - OBJECTIVES: Fat in the lower body is not associated with the same risk of cardiovascular disease as fat in the upper body. Is this explained by differences in the physiological functioning of the two depots? This study had two objectives: 1) to determine whether fat mobilization and blood flow differ between gluteal and abdominal adipose tissues in humans, and 2) to develop a new technique to assess gluteal adipose tissue function directly. RESEARCH METHODS AND PROCEDURES: We performed detailed in vivo studies of adipose tissue function involving the assessment of fat mobilization by measurement of adipose tissue blood flows, arterio-venous differences of metabolites across each depot, and gene expression in tissue biopsies in a small-scale physiological study. RESULTS: Gluteal adipose tissue has a lower blood flow (67% lower, p < 0.05) and lower hormone-sensitive lipase rate of action (87% lower, p < 0.05) than abdominal adipose tissue. Lipoprotein lipase rate of action and mRNA expression are not different between the depots. This is the first demonstration of a novel technique to directly investigate gluteal adipose tissue metabolism. DISCUSSION: Direct assessment of fasting adipose tissue metabolism in defined depots show that the buttock is metabolically "silent" in terms of fatty acid release compared with the abdomen. PMID- 14742850 TI - Plasma adiponectin levels and metabolic factors in nondiabetic adolescents. AB - OBJECTIVES: The relationship of plasma adiponectin levels with various anthropometric and metabolic factors has been surveyed extensively in adults. However, how plasma adiponectin levels are related to various anthropometric indices and cardiovascular risk factors in adolescents is not as vigorously studied. In this study, we investigated this among healthy nondiabetic adolescents. RESEARCH METHODS AND PROCEDURES: Two hundred thirty nondiabetic subjects (125 boys and 105 girls, approximately 10 to 19 years old) were included. The plasma adiponectin, fasting plasma glucose, insulin, lipids and anthropometric indices including body height, weight, waist circumference, and hip circumference were examined. Body fat mass (FM) and percentage were obtained from DXA scan. The homeostasis model assessment was applied to estimate the degree of insulin resistance. RESULTS: The plasma adiponectin levels were significantly higher in girls (30.79 +/- 14.48 micro g/mL) than boys (22.87 +/- 11.41 micro g/mL). The plasma adiponectin levels were negatively related to BMI, FM, FM percentage, waist circumference, waist-to-hip ratio, insulin resistance, plasma insulin, triglycerides, and uric acid levels, but positively with high density lipoprotein cholesterol (HDL-C) with the adjustment for age and gender. Using different multivariate linear regression models, only age and HDL-C were consistently related to the plasma adiponectin levels after adjustment for the other variables. DISCUSSION: The relationship between plasma adiponectin and various anthropometric indices and metabolic factors, especially HDL-C, previously reported in adults was present in the healthy nondiabetic adolescents. Whether variation of plasma adiponectin levels in healthy nondiabetic adolescents may influence their future coronary artery disease risk warrants further investigation. PMID- 14742851 TI - beta(3) Adrenergic receptor polymorphism and obesity-related phenotypes in hypertensive patients. AB - OBJECTIVES: Obesity is a complex trait that is affected by both environmental and genetic risk factors. The beta(3) adrenergic receptor (ADRB3) is expressed in adipose tissue and plays a role in energy metabolism. A missense mutation on codon 64 of this gene (W64R) is associated with receptor malfunction. Previous studies examining the relation between this polymorphism and obesity produced inconsistent findings. The current study assessed the association between the W64R genotype and obesity-related phenotypes, including body weight, BMI, and serum triglycerides, cholesterol, and glucose. RESEARCH METHODS AND PROCEDURES: We determined the ADRB3 W64R genotypes and fasting serum lipid and glucose concentrations for 695 hypertensive adults (336 men,359 women) from a rural county in Anhui Province, China. Multivariate linear regression models were fit to detect associations between the genetic polymorphism and obesity-related phenotypes. RESULTS: The ADRB3 W64R polymorphism was significantly associated with body weight and BMI in men but not in women. After controlling for potential confounding variables, men who were homozygous for the R64 allele were 11.8 kg heavier (p < 0.001) and had a BMI that was 3.7 kg/m(2) greater (p = 0.001) than men who were homozygous for the W64 allele. Serum concentrations of lipids and glucose were found not associated with the genetic polymorphism. DISCUSSION: The ADRB3 R64 allele was associated with increased body weight and BMI in men but not in women. The genetic association was not modified by triglyceride, cholesterol, blood glucose, or blood pressure levels of the subjects. PMID- 14742852 TI - Sexual dimorphism in the response of adipose mass and cellularity to graded caloric restriction. AB - OBJECTIVE: To investigate the effects of mild to moderate caloric restriction on parameters of body growth, fat mass, and adipose tissue cellularity in female and male Wistar rats. RESEARCH METHODS AND PROCEDURES: Three-month-old female and male Wistar rats were subjected to a chronic, mild to moderate caloric restriction paradigm (5%, 10%, or 20% reduction in caloric intake from ad libitum values) for 6 months. This was accomplished using a unique automated feeder system tailored to the food consumption levels of individual rats. Body weight and length, weight of lean organs, regional adipose mass, and adipose cellularity were measured before and after the diet restriction. RESULTS: Caloric restriction produced proportional decelerations in body weight increases in both genders, without significant changes in body length or lean organ mass. Marked and disproportional reductions in regional adipose tissue mass were produced at all levels of food restriction (even at 5% restriction). An unexpected finding was that in response to graded caloric restriction, female rats preserved adipose fat cell number at the expense of fat cell volume, whereas the converse was seen for male rats. DISCUSSION: These studies demonstrate a sexual dimorphism in the response to mild to moderate degrees of chronic caloric restriction. At low levels of caloric restriction, it is possible to affect regional adipose mass and cellularity while preserving lean organ mass. PMID- 14742854 TI - Evaluation of a quantitative magnetic resonance method for mouse whole body composition analysis. AB - OBJECTIVE: To evaluate applicability, precision, and accuracy of a new quantitative magnetic resonance (QMR) analysis for whole body composition of conscious live mice. RESEARCH METHODS AND PROCEDURES: Repeated measures of body composition were made by QMR, DXA, and classic chemical analysis of carcass using live and dead mice with different body compositions. Caloric lean and dense diets were used to produce changes in body composition. In addition, different strains of mice representing widely diverse populations were analyzed. RESULTS: Precision was found to be better for QMR than for DXA. The coefficient of variation for fat ranged from 0.34% to 0.71% compared with 3.06% to 12.60% for DXA. Changes in body composition in response to dietary manipulation were easily detected using QMR. An increase in fat mass of 0.6 gram after 1 week (p < 0.01) was demonstrated in the absence of hyperphagia or a change in mean body weight. DISCUSSION: QMR and DXA detected similar fat content, but the improved precision afforded by QMR compared with DXA and chemical analysis allowed detection of a significant difference in body fat after 7 days of consuming a diet rich in fat even though average body weight did not significantly change. QMR provides a very precise, accurate, fast, and easy-to-use method for determining fat and lean tissue of mice without the need for anesthesia. Its ability to detect differences with great precision should be of value when characterizing phenotype and studying regulation of body composition brought about by pharmacological and dietary interventions in energy homeostasis. PMID- 14742853 TI - Restrained eating behavior and the metabolic response to dietary energy restriction in women. AB - OBJECTIVE: To determine whether prior eating behavior characterized by dietary restraint alters responses in energy expenditure and substrate oxidation associated with a short-term, energy-restricted diet. RESEARCH METHODS AND PROCEDURES: A repeated-measures, 3-day diet-intervention study of adequate (125 kJ/kg of body weight) or restricted (62.5 kJ/kg) energy intake was conducted with 30 women, 20 to 46 years, BMI 25 to 45 kg/m(2), whose prior eating behavior was "restrained" or "unrestrained." The Eating Inventory (cognitive restraint subscale) was used to measure restrained eating behavior. Energy expenditure and substrate oxidation were measured after a 12-hour fast and during the first and fourth hours after a standard meal. Plasma glucose, nonesterified fatty acids, and insulin were measured at corresponding times. Body composition was determined by total body electrical conductivity. RESULTS: Resting energy expenditure was not affected by 3 days of energy restriction. Short-term energy restriction resulted in lower respiratory-exchange ratios, higher rates of fat oxidation, and lower rates of carbohydrate oxidation. Subjects classified as restrained eaters had higher postprandial respiratory-exchange ratios and carbohydrate-oxidation rates compared with unrestrained eaters. Fasting insulin concentrations were lower in restrained eaters. These effects associated with prior eating behavior were independent of the diet intervention. DISCUSSION: Metabolic outcomes associated with a 3-day energy-restricted diet (i.e., increased fat oxidation and decreased carbohydrate oxidation) were not affected by prior restrained eating behavior. However, restrained eating behavior was associated with increased carbohydrate oxidation after a mixed meal. This effect of restrained eating behavior may be attributable to increased insulin sensitivity. PMID- 14742855 TI - Weight counseling patterns of U. S. Pediatricians. AB - OBJECTIVE: To estimate the proportion and characteristics of U.S. pediatricians who reportedly counsel their patients about maintaining a healthy weight. RESEARCH METHODS AND PROCEDURES: Beginning in October 1998, information was collected from 813 primary care pediatricians randomly selected from a nationally representative sample. Pediatricians were asked how frequently they counseled about maintaining a healthy weight during the well-care visits of patients in three age groups. Multivariable logistic regression determined which physician characteristics were associated with counseling. RESULTS: Approximately fifty percent of pediatricians reportedly always counseled about maintaining a healthy weight. Those who always counseled were more likely to be women, to spend more time with patients during well-care visits, and to conduct more well-care visits per week from patients in one particular age group. Most pediatricians who responded that they always counseled about healthy weight reported that they counseled about physical activity and nutrition, but not about balancing caloric intake with expenditure. DISCUSSION: Although many pediatricians report counseling about healthy weight, the frequency of counseling might be further increased by increasing the amount of time the patient spends during office visits with the pediatrician or with other professional staff, such as nurses or dietitians. PMID- 14742865 TI - Rapid detection of single nucleotide polymorphisms associated with spinal muscular atrophy by use of a reusable fibre-optic biosensor. AB - Rapid (<2 min) and quantitative genotyping for single nucleotide polymorphisms (SNPs) associated with spinal muscular atrophy was done using a reusable (approximately 80 cycles of application) fibre-optic biosensor over a clinically relevant range (0-4 gene copies). Sensors were functionalized with oligonucleotide probes immobilized at high density (approximately 7 pmol/cm2) to impart enhanced selectivity for SNP discrimination and used in a total internal reflection fluorescence detection motif to detect 202 bp PCR amplicons from patient samples. Real-time detection may be done over a range of ionic strength conditions (0.1-1.0 M) without stringency rinsing to remove non-selectively bound materials and without loss of selectivity, permitting a means for facile sample preparation. By using the time-derivative of fluorescence intensity as the analytical parameter, linearity of response may be maintained while allowing for significant reductions in analysis time (10-100-fold), permitting for the completion of measurements in under 1 min. PMID- 14742867 TI - Division of labor among distinct subtypes of inhibitory neurons in a cortical microcircuit of working memory. AB - A conspicuous feature of cortical organization is the wide diversity of inhibitory interneurons; their differential computational functions remain unclear. Here we propose a local cortical circuit in which three major subtypes of interneurons play distinct roles. In a model designed for spatial working memory, stimulus tuning of persistent activity arises from the concerted action of widespread inhibition mediated by perisoma-targeting (parvalbumin-containing) interneurons and localized disinhibition of pyramidal cells via interneuron targeting (calretinin-containing) interneurons. Moreover, resistance against distracting stimuli (a fundamental property of working memory) is dynamically controlled by dendrite-targeting (calbindin-containing) interneurons. The experimental observation of inverted tuning curves of monkey prefrontal neurons recorded during working memory supports a key model prediction. This work suggests a framework for understanding the division of labor and cooperation among different inhibitory cell types in a recurrent cortical circuit. PMID- 14742866 TI - Antiinflammatory activity of soluble guanylate cyclase: cGMP-dependent down regulation of P-selectin expression and leukocyte recruitment. AB - Nitric oxide (NO) production by the vascular endothelium maintains an essential antiinflammatory, cytoprotective influence on the blood vessel wall. A key component of this activity is attributed to prevention of leukocyte-endothelial cell interactions, yet the underlying mechanisms remain unclear. The NO receptor, soluble guanylate cyclase (sGC), is expressed in endothelial cells but fulfils an unknown function. Therefore, we used intravital microscopy in mesenteric postcapillary venules from WT and endothelial nitric oxide synthase (eNOS) knockout (eNOS(-/-)) mice, and an sGC activator (BAY 41-2272), to investigate a potential role for sGC in the regulation of adhesion molecule expression and leukocyte recruitment. Leukocyte rolling and adhesion was 6-fold greater in eNOS( /-) than WT animals. BAY 41-2272 and the NO-donor, diethylamine-NONOate, reduced leukocyte rolling and adhesion in eNOS(-/-) mice to levels observed in WT animals. These effects were blocked by the sGC inhibitor ODQ [1H (1,2,4)oxadiazolo(4,3-a)quinoxalin-1-one], which itself caused a 6-fold increase in leukocyte rolling and adhesion in WT mice. Increased leukocyte rolling and adhesion in IL-1beta-treated mice was also inhibited by BAY 41-2272. Fluorescence activated cell sorting analysis in vitro and a specific P-selectin neutralizing antibody in vivo revealed that selective down-regulation of P-selectin expression accounted for the antiadhesive effects of sGC activation. These data demonstrate that sGC plays a key antiinflammatory role by inhibiting P-selectin expression and leukocyte recruitment. PMID- 14742869 TI - Mapping knowledge domains. PMID- 14742868 TI - An effective second-generation outer surface protein A-derived Lyme vaccine that eliminates a potentially autoreactive T cell epitope. AB - The antigenic component of a common Lyme disease vaccine is recombinant outer surface protein A (rOspA) of Borrelia burgdorferi (Bb), the causative agent of Lyme disease. Coincidentally, patients with chronic, treatment-resistant Lyme arthritis develop an immune response against OspA, whereas those with acute Lyme disease usually do not. Treatment-resistant Lyme arthritis occurs in a subset of Lyme arthritis patients and is linked to HLA.DRB1*0401 (DR4) and related alleles. Recent work from our laboratory identified T cell crossreactivity between epitopes of OspA and lymphocyte function-associated antigen 1alpha(L) chain (LFA 1alpha(L)) in these patients. We generated a form of rOspA, FTK-OspA, in which the LFA-1alpha(L)/rOspA crossreactive T cell epitope was mutated to reduce the possible risk of autoimmunity in genetically susceptible individuals. FTK-OspA did not stimulate human or mouse DR4-restricted, WT-OspA-specific T cells, whereas it did stimulate antibody responses specific for WT-OspA that were similar to mice vaccinated WT-OspA. We show here that the protective efficacy of FTK-OspA is indistinguishable from that of WT-OspA in vaccination trials, as both C3H/HeJ and BALB/c FTK-OspA-vaccinated mice were protected from Bb infection. These data demonstrate that this rOspA-derived vaccine lacking the predicted cross-reactive T cell epitope, but retaining the capacity to elicit antibodies against infection, is effective in generating protective immunity. PMID- 14742870 TI - Alternative splicing generates an isoform of the human Sef gene with altered subcellular localization and specificity. AB - Receptor tyrosine kinases (RTKs) control a multitude of biological processes and are therefore subjected to multiple levels of regulation. Negative feedback is one of the mechanisms that provide an effective means to control RTK-mediated signaling. Sef has recently been identified as a specific antagonist of fibroblast growth factor (FGF) signaling in zebrafish and subsequently in mouse and human. Sef encodes a putative type I transmembrane protein that antagonizes the Ras/mitogen-activated protein kinase pathway in all three species. Mouse Sef was also shown to inhibit the phosphatidylinositol 3-kinase pathway. We show here that an alternative splicing mechanism generates an isoform of human Sef, hSef-b, which unlike the previously reported Sef (hSef-a) is a cytosolic protein. Contrary to hSef-a, which is ubiquitously expressed, hSef-b transcripts display a restricted pattern of expression in human tissues. hSef-b inhibits FGF-induced cell proliferation and prevents the activation of mitogen-activated protein kinase without affecting the upstream component MAPK kinase. Furthermore, hSef-b does not antagonize FGF induction of the phosphatidylinositol 3-kinase pathway. In addition to the effects on FGF signaling, hSef-b inhibited cellular response to platelet-derived growth factor but not other RTK ligands. Therefore, alternative splicing of the hSef gene expands the Sef feedback inhibition repertoire of RTK signaling. PMID- 14742871 TI - Convergent evolution of disease resistance gene specificity in two flowering plant families. AB - Plant disease resistance (R) genes that mediate recognition of the same pathogen determinant sometimes can be found in distantly related plant families. This observation implies that some R gene alleles may have been conserved throughout the diversification of land plants. To address this question, we have compared R genes from Glycine max (soybean), Rpg1-b, and Arabidopsis thaliana, RPM1, that mediate recognition of the same type III effector protein from Pseudomonas syringae, AvrB. RPM1 has been cloned previously, and here, we describe the isolation of Rpg1-b. Although RPM1 and Rpg1-b both belong to the coiled-coil nucleotide binding site (NBS) Leu-rich repeat (LRR) class of R genes, they share only limited sequence similarity outside the conserved domains characteristic of this class. Phylogenetic analyses of A. thaliana and legume NBS-LRR sequences demonstrate that Rpg1-b and RPM1 are not orthologous. We conclude that convergent evolution, rather than the conservation of an ancient specificity, is responsible for the generation of these AvrB-specific genes. PMID- 14742872 TI - The WRKY70 transcription factor: a node of convergence for jasmonate-mediated and salicylate-mediated signals in plant defense. AB - Cross talk between salicylic acid (SA)- and jasmonic acid (JA)-dependent defense signaling has been well documented in plants, but how this cross talk is executed and the components involved remain to be elucidated. We demonstrate that the plant-specific transcription factor WRKY70 is a common component in SA- and JA mediated signal pathways. Expression of WRKY70 is activated by SA and repressed by JA. The early induction of WRKY70 by SA is NPR1-independent, but functional NPR1 is required for full-scale induction. Epistasis analysis suggested that WRKY70 is downstream of NPR1 in an SA-dependent signal pathway. Modulation of WRKY70 transcript levels by constitutive overexpression increases resistance to virulent pathogens and results in constitutive expression of SA-induced pathogenesis-related genes. Conversely, antisense suppression of WRKY70 activates JA-responsive/COI1-dependent genes. The effect of WRKY70 is not caused by subsequent changes in SA or JA levels. We suggest that WRKY70 acts as an activator of SA-induced genes and a repressor of JA-responsive genes, integrating signals from these mutually antagonistic pathways. PMID- 14742873 TI - Aux/IAA proteins contain a potent transcriptional repression domain. AB - Aux/IAA proteins are short-lived nuclear proteins that repress expression of primary/early auxin response genes in protoplast transfection assays. Repression is thought to result from Aux/IAA proteins dimerizing with auxin response factor (ARF) transcriptional activators that reside on auxin-responsive promoter elements, referred to as AuxREs. Most Aux/IAA proteins contain four conserved domains, designated domains I, II, III, and IV. Domain II and domains III and IV play roles in protein stability and dimerization, respectively. A clear function for domain I had not been established. Results reported here indicate that domain I in Aux/IAA proteins is an active repression domain that is transferable and dominant over activation domains. An LxLxL motif within domain I is important for conferring repression. The dominance of Aux/IAA repression domains over activation domains in ARF transcriptional activators provides a plausible explanation for the repression of auxin response genes via ARF-Aux/IAA dimerization on auxin-responsive promoters. PMID- 14742874 TI - Apoplastic synthesis of nitric oxide by plant tissues. AB - Nitric oxide (NO) is an important signaling molecule in animals and plants. In mammals, NO is produced from Arg by the enzyme NO synthase. In plants, NO synthesis from Arg using an NO synthase-type enzyme and from nitrite using nitrate reductase has been demonstrated previously. The data presented in this report strongly support the hypothesis that plant tissues also synthesize NO via the nonenzymatic reduction of apoplastic nitrite. As measured by mass spectrometry or an NO-reactive fluorescent probe, Hordeum vulgare (barley) aleurone layers produce NO rapidly when nitrite is added to the medium in which they are incubated. NO production requires an acid apoplast and is accompanied by a loss of nitrite from the medium. Phenolic compounds in the medium can increase the rate of NO production. The possible significance of apoplastic NO production for germinating grain and for plant roots is discussed. PMID- 14742875 TI - The Arabidopsis thaliana ABSCISIC ACID-INSENSITIVE8 encodes a novel protein mediating abscisic acid and sugar responses essential for growth. AB - Abscisic acid (ABA) regulates many aspects of plant growth and development, yet many ABA response mutants present only subtle phenotypic defects, especially in the absence of stress. By contrast, the ABA-insensitive8 (abi8) mutant, isolated on the basis of ABA-resistant germination, also displays severely stunted growth, defective stomatal regulation, altered ABA-responsive gene expression, delayed flowering, and male sterility. The stunted growth of the mutant is not rescued by gibberellin, brassinosteroid, or indoleacetic acid application and is not attributable to excessive ethylene response, but supplementing the medium with Glc improves viability and root growth. In addition to exhibiting Glc-dependent growth, reflecting decreased expression of sugar-mobilizing enzymes, abi8 mutants are resistant to Glc levels that induce developmental arrest of wild-type seedlings. Studies of genetic interactions demonstrate that ABA hypersensitivity conferred by the ABA-hypersensitive1 mutation or overexpression of ABI3 or ABI5 does not suppress the dwarfing and Glc dependence caused by abi8 but partially suppresses ABA-resistant germination. By contrast, the ABA-resistant germination of abi8 is epistatic to the hypersensitivity caused by ethylene-insensitive2 (ein2) and ein3 mutations, yet ABI8 appears to act in a distinct Glc response pathway from these EIN loci. ABI8 encodes a protein with no domains of known function but belongs to a small plant-specific protein family. Database searches indicate that it is allelic to two dwarf mutants, elongation defective1 and kobito1, previously shown to disrupt cell elongation, cellulose synthesis, vascular differentiation, and root meristem maintenance. The cell wall defects appear to be a secondary effect of the mutations because Glc treatment restores root growth and vascular differentiation but not cell elongation. Although the ABI8 transcript accumulates in all tested plant organs in both wild-type and ABA response mutants, an ABI8-beta-glucuronidase fusion protein is localized primarily to the elongation zone of roots, suggesting substantial post transcriptional regulation of ABI8 accumulation. This localization pattern is sufficient to complement the mutation, indicating that ABI8 acts either at very low concentrations or over long distances within the plant body. PMID- 14742877 TI - Mutations in the pale aleurone color1 regulatory gene of the Zea mays anthocyanin pathway have distinct phenotypes relative to the functionally similar TRANSPARENT TESTA GLABRA1 gene in Arabidopsis thaliana. AB - The pale aleurone color1 (pac1) locus, required for anthocyanin pigment in the aleurone and scutellum of the Zea mays (maize) seed, was cloned using Mutator transposon tagging. pac1 encodes a WD40 repeat protein closely related to anthocyanin regulatory proteins ANTHOCYANIN11 (AN11) (Petunia hybrida [petunia]) and TRANSPARENT TESTA GLABRA1 (TTG1) (Arabidopsis thaliana). Introduction of a 35S-Pac1 transgene into A. thaliana complemented multiple ttg1 mutant phenotypes, including ones nonexistent in Z. mays. Hybridization of Z. mays genomic BAC clones with the pac1 sequence identified an additional related gene, mp1. PAC1 and MP1 deduced protein sequences were used as queries to build a phylogenetic tree of homologous WD40 repeat proteins, revealing an ancestral gene duplication leading to two clades in plants, the PAC1 clade and the MP1 clade. Subsequent duplications within each clade have led to additional WD40 repeat proteins in particular species, with all mutants defective in anthocyanin expression contained in the PAC1 clade. Substantial differences in pac1, an11, and ttg1 mutant phenotypes suggest the evolutionary divergence of regulatory mechanisms for several traits that cannot be ascribed solely to divergence of the dicot and monocot protein sequences. PMID- 14742876 TI - Downregulation of the Petunia hybrida alpha-expansin gene PhEXP1 reduces the amount of crystalline cellulose in cell walls and leads to phenotypic changes in petal limbs. AB - The expansins comprise a family of proteins that appear to be involved in the disruption of the noncovalent bonds between cellulose microfibrils and cross linking glycans, thereby promoting wall creep. To understand better the expansion process in Petunia hybrida (petunia) flowers, we isolated a cDNA corresponding to the PhEXP1 alpha-expansin gene of P. hybrida. Evaluation of the tissue specificity and temporal expression pattern demonstrated that PhEXP1 is preferentially expressed in petal limbs during development. To determine the function of PhEXP1, we used a transgenic antisense approach, which was found to cause a decrease in petal limb size, a reduction in the epidermal cell area, and alterations in cell wall morphology and composition. The diminished cell wall thickness accompanied by a reduction in crystalline cellulose indicates that the activity of PhEXP1 is associated with cellulose metabolism. Our results suggest that expansins play a role in the assembly of the cell wall by affecting either cellulose synthesis or deposition. PMID- 14742878 TI - Two classes of the CDh1-type activators of the anaphase-promoting complex in plants: novel functional domains and distinct regulation. AB - The Cdc20 and Cdh1/Fzr proteins are the substrate-specific activators of the anaphase-promoting complex (APC). In Medicago truncatula, the MtCcs52A and MtCcs52B proteins represent two subgroups of the Cdh1-type activators, which display differences in their cell cycle regulation, structure, and function. The ccs52A transcripts are present in all phases of the cell cycle. By contrast, expression of ccs52B is restricted to late G2-phase and M-phase, and its induced overexpression in BY2 cells inhibited mitosis. MtCcs52A is active in Schizosaccharomyces pombe and binds to the S. pombe APC, whereas MtCcs52B does not because of differences in the N-terminal region. We identified a new functional domain, the Cdh1-specific motif conserved in the Cdh1 proteins that, in addition to the C-box and the terminal Ile and Arg residues, was essential for the activity and required for efficient binding to the APC. Moreover, we demonstrate that cyclin-dependent kinase phosphorylation sites adjacent to the C box may regulate the interaction with the APC. In the different plant organs, the expression of Mtccs52A and Mtccs52B displayed differences and indicated the involvement of the APC in differentiation processes. PMID- 14742879 TI - Transgenic evaluation of activated mutant alleles of SOS2 reveals a critical requirement for its kinase activity and C-terminal regulatory domain for salt tolerance in Arabidopsis thaliana. AB - In Arabidopsis thaliana, the calcium binding protein Salt Overly Sensitive3 (SOS3) interacts with and activates the protein kinase SOS2, which in turn activates the plasma membrane Na(+)/H(+) antiporter SOS1 to bring about sodium ion homeostasis and salt tolerance. Constitutively active alleles of SOS2 can be constructed in vitro by changing Thr(168) to Asp in the activation loop of the kinase catalytic domain and/or by removing the autoinhibitory FISL motif from the C-terminal regulatory domain. We expressed various activated forms of SOS2 in Saccharomyces cerevisiae (yeast) and in A. thaliana and evaluated the salt tolerance of the transgenic organisms. Experiments in which the activated SOS2 alleles were coexpressed with SOS1 in S. cerevisiae showed that the kinase activity of SOS2 is partially sufficient for SOS1 activation in vivo, and higher kinase activity leads to greater SOS1 activation. Coexpression of SOS3 with SOS2 forms that retained the FISL motif resulted in more dramatic increases in salt tolerance. In planta assays showed that the Thr(168)-to-Asp-activated mutant SOS2 partially rescued the salt hypersensitivity in sos2 and sos3 mutant plants. By contrast, SOS2 lacking only the FISL domain suppressed the sos2 but not the sos3 mutation, whereas truncated forms in which the C terminus had been removed could not restore the growth of either sos2 or sos3 plants. Expression of some of the activated SOS2 proteins in wild-type A. thaliana conferred increased salt tolerance. These studies demonstrate that the protein kinase activity of SOS2 is partially sufficient for activation of SOS1 and for salt tolerance in vivo and in planta and that the kinase activity of SOS2 is limiting for plant salt tolerance. The results also reveal an essential in planta role for the SOS2 C-terminal regulatory domain in salt tolerance. PMID- 14742880 TI - Peroxisomal Acyl-CoA synthetase activity is essential for seedling development in Arabidopsis thaliana. AB - In plants and other eukaryotes, long-chain acyl-CoAs are assumed to be imported into peroxisomes for beta-oxidation by an ATP binding cassette (ABC) transporter. However, two genes in Arabidopsis thaliana, LACS6 and LACS7, encode peroxisomal long-chain acyl-CoA synthetase (LACS) isozymes. To investigate the biochemical and biological roles of peroxisomal LACS, we identified T-DNA knockout mutants for both genes. The single-mutant lines, lacs6-1 and lacs7-1, were indistinguishable from the wild type in germination, growth, and reproductive development. By contrast, the lacs6-1 lacs7-1 double mutant was specifically defective in seed lipid mobilization and required exogenous sucrose for seedling establishment. This phenotype is similar to the A. thaliana pxa1 mutants deficient in the peroxisomal ABC transporter and other mutants deficient in beta oxidation. Our results demonstrate that peroxisomal LACS activity and the PXA1 transporter are essential for early seedling growth. The peroxisomal LACS activity would be necessary if the PXA1 transporter delivered unesterified fatty acids into the peroxisomal matrix. Alternatively, PXA1 and LACS6/LACS7 may act in parallel pathways that are both required to ensure adequate delivery of acyl-CoA substrates for beta-oxidation and successful seedling establishment. PMID- 14742881 TI - Official positions of the International Society for Clinical Densitometry. AB - The International Society for Clinical Densitometry (ISCD) has convened two Position Development Conferences at which a panel of experts agreed on recommendations for performance and clinical applications of bone density testing. These recommendation were reviewed by the ISCD Board of Directors, and those approved by the board are now official positions of the ISCD. These include (1) indications for bone density testing, (2) reference databases for T-scores, (3) standards for performing central dual-energy X-ray absorptiometry (DXA) for diagnosis, (4) interpretation of peripheral bone density results, (5) diagnosis of osteoporosis in postmenopausal women, (6) diagnosis of osteoporosis in men, (7) diagnosis in premenopausal women, (8) diagnosis in children, (9) indications and interpretation for serial bone mass measurement, (10) technical standards for phantom scanning and calibration, (11) technical standards for cross-calibration of DXAsystems, and (12) standards for reporting of bone density results including correct nomenclature and preferred number of decimal digits. PMID- 14742882 TI - Position statement : executive summary. The Writing Group for the International Society for Clinical Densitometry (ISCD) Position Development Conference. AB - The International Society for Clinical Densitometry (ISCD) held a Position Development Conference in July 2003, at which time positions developed and researched by the organization's Scientific Advisory Committee were presented to a panel of international experts in the field of bone density testing. This panel reached agreement on a series of positions that were subsequently approved by the Board of Directors of the ISCD and are now official policy of the ISCD. These positions, which are outlined in this article and discussed in greater detail in subsequent articles in this journal, include (1) affirmation of the use of the World Health Organization classification for the diagnosis of osteoporosis in postmenopausal women; (2) the diagnosis of osteoporosis in men; (3) the diagnosis of osteoporosis in premenopausal women; (4) the diagnosis of osteoporosis in children; (5) technical standards for skeletal regions of interest by dual-energy X-ray absorptiometry (DXA); (6) the use of new technologies, such as vertebral fracture assessment; (7) technical standards for quality assurance, including phantom scanning and calibration; (8) technical standards for the performance of precision assessment at bone density testing centers, and for cross-calibration of DXA devices; (9) indications for bone density testing; (10) appropriate information for a bone density report; and (11) nomenclature and decimal places for bone density reporting. PMID- 14742883 TI - Position statement: introduction, methods, and participants. The Writing Group for the International Society for Clinical Densitometry (ISCD) Position Development Conference. AB - Following publication of the proceedings from the first Position Development Conference (PDC) of the International Society for Clinical Densitometry (ISCD), members of the ISCD Scientific Advisory Committee (SAC) addressed additional topics of interest in the field of bone densitometry. These topics were addressed at a subsequent PDC, which was held in Cincinnati, Ohio, July 25-27, 2003. Five topics were chosen for discussion: (1) the diagnosis of osteoporosis in men, premenopausal women, and children; (2) technical standardization for dual-energy X-ray absorptiometry (DXA); (3) indications for bone densitometry; (4) reporting of bone density results; and (5) nomenclature and decimal places for bone densitometry. This report describes the methodology used for the development, presentation, and finalization of PDC positions. These positions are discussed in the following papers. PMID- 14742884 TI - Diagnosis of osteoporosis in men, premenopausal women, and children. AB - The World Health Organization (WHO) classification of bone mineral density is based on population studies in postmenopausal women. However, the increased use of bone densitometry has raised questions regarding the applicability of this classification to men, premenopausal women, and children. These questions were addressed at the International Society for Clinical Densitometry 2003 Position Development Conference. T-scores can be used and a diagnosis of osteoporosis made for T-scores of -2.5 or less (male reference database) in men age 65 yr and older and in men from 50 to 64 yr of age if other risk factors for fracture are present. The WHO classification should not be applied to premenopausal women. Men and premenopausal women with secondary causes of low bone density or bone loss may be clinically diagnosed with osteoporosis, taking bone density into consideration. In children and adolescents (males and females less than 20 yr of age), there are no densitometric criteria for diagnosing osteoporosis. If Z scores are -2.0 or less (using pediatric databases of age-matched controls), then a characterization such as "low bone density for chronologic age" is appropriate. In men under 50 yr of age, premenopausal women, and children, Z-scores, not T scores, should be used when reporting bone density results. PMID- 14742885 TI - Technical standardization for dual-energy x-ray absorptiometry. AB - Clinical applications of dual-energy X-ray absorptiometry (DXA) should be updated periodically to address new technologies and inconsistencies in the use of existing technologies. The 2003 Position Development Conference of the International Society for Clinical Densitometry addressed issues regarding hip and forearm regions of interest (ROIs), vertebral fracture assessment (VFA), quality assurance (QA), precision, and cross-calibration. Data are insufficient to determine whether the mean T-score of both hips can be used for diagnosis. However, the mean bone mineral density of both hips can be used for monitoring. The 33% radius is the preferred forearm ROI for the diagnosis of osteoporosis. Data are insufficient to determine indications and clinical utility of VFA. Minimum standards for a QA program were established. Phantom scanning should be performed at least once per week. Precision assessment should use patients representative of a DXA center's patient population. If a center has more than one technologist, then the average precision may be used. Repeat precision assessment is necessary if a new DXA system is installed or a technologist's skills have changed. Knowledge of precision is critical to DXA interpretation, and approval from an institutional review board should not be required. Practical standardized procedures for cross-calibration of DXA systems should be developed. PMID- 14742886 TI - Indications and reporting for dual-energy x-ray absorptiometry. AB - Although dual-energy X-ray absorptiometry (DXA) is widely used to measure bone mineral density (BMD), there are few guidelines regarding indications for testing in populations other than postmenopausal women or for reporting the results from DXA studies. At the 2003 Position Development Conference of the International Society for Clinical Densitometry, positions were developed to formalize indications for bone density testing and the components of a DXA report. Bone density testing is indicated for the following individuals: women 65 yr of age and older, postmenopausal women under age 65 yr with risk factors, men 70 yr of age and older, adults with fragility fracture, adults taking a medication or adults with a disease or condition associated with low bone mass or bone loss, any individual being treated for low bone mass to monitor treatment effect, and any individual in whom evidence of bone loss would affect treatment decisions. The DXA report should include patient identification and demographics, identification of the referring provider, indications for the test, an explanation of the technique, and abnormalities in performing the study. In addition, the report should include BMD values, the T-score and/or Z-score as appropriate, World Health Organization (WHO) classification for diagnosis, risk factors, and recommendations for follow-up. PMID- 14742887 TI - Nomenclature and decimal places in bone densitometry. AB - There are inconsistencies in densitometry nomenclature and in the use of decimal places in reporting bone density results. These issues were discussed at the 2003 Position Development Conference (PDC) of the International Society for Clinical Densitometry (ISCD). Regarding nomenclature, there was agreement that DXA is the preferred designation for dual-energy X-ray absorptiometry. T-score (not T score, t score, or tscore) is the preferred designation for the standard deviation variance of bone mineral density (BMD) from the mean BMD of the young adult reference population. Z-score terminology should be consistent with that for T score. For the reporting of bone densitometry results, BMD should be expressed to three decimal places, bone mineral content and area to two decimal places, T scores and Z-scores to one decimal place, and percentages (percentage of reference population mean BMD) as integers. PMID- 14742888 TI - Standards and guidelines for performing central dual-energy x-ray absorptiometry in premenopausal women, men, and children. AB - The Canadian Panel of the International Society for Clinical Densitometry has developed standards in order to establish the minimum level of acceptable performance for the practice of bone densitometry in Canada. Previously, this group addressed the performance of densitometry in postmenopausal women. This report addresses the use of densitometry in men, premenopausal women, and children with a focus on dual-energy X-ray absorptiometry. PMID- 14742889 TI - Height loss and osteoporosis of the hip. AB - Osteoporosis of the hip is associated with hip fracture, a devastating outcome on both an individual and aggregate basis. Height loss is a frequent manifestation of vertebral osteoporosis and is simple to evaluate in the clinical setting. The goal of this study was to determine whether height loss is significantly associated with low bone mineral density at the femur, using a retrospective review of cross-sectional data from 2108 women referred for a bone density scan. Collected data included self-reported maximum adult height, current height, and total hip bone mineral density, along with other demographic and risk factor information. We then investigated the relationship between height loss and osteoporosis using multinomial logistic regression modeling. We found that height loss of 2 in. or more is a highly significant predictor of osteoporosis at the hip. In particular, the odds women had osteoporosis at the hip, as determined by total hip bone mineral density, increased 4.4 times (95% confidence interval, 2.6 7.4) if the women had lost > or = 2 but < 3 in. of height. In addition, women with at least 3 in. of height loss had odds of osteoporosis of the hip that were 9.6 times greater (95% confidence interval, 4.8-19.2) than women with less than an inch of height loss. These odds ratios were adjusted for the confounding variables of age, weight, and maximum adult height. Our findings suggest loss of height may be an important clue in detecting osteoporosis of the hip, implying that evaluation of height loss should be routine in the outpatient setting. PMID- 14742890 TI - Using mobile DXA to improve access to osteoporosis care: unit design, program development, implementation, and outcomes. AB - Osteoporosis diagnosis and monitoring is best accomplished with dual X-ray absorptiometry (DXA), but technology availability can hinder access to care. We designed a mobile DXA program incorporating a Hologic Delphi-C trade mark bone densitometer housed in a specially configured 30-ft Winnebago trade mark. The mobile DXA program provided osteoporosis testing and education at the convenience of the patient's primary care site within our rural health care system. DXA results were sent electronically to the patient's physician within 48-72 h. The mobile DXA patient group tended to be older and at high risk for future fracture. The service provided was rated as excellent by patients. Given the volume of patients studied, the program was financially self-sustaining. Other healthcare systems or groups should consider development of a similar program. PMID- 14742891 TI - Evaluation of an osteoporosis self-referral program to enhance management outcomes. AB - Osteoporosis is a common but undertreated condition. While bone density is known to predict fracture risk, there is currently no economical way of measuring general population risk. This study examined whether an economical screening technique could improve diagnosis and lead to appropriate outcomes in the management of osteoporosis. A self-referral program was established to provide women with information about osteoporosis and an individualized risk assignment. A high-risk designation was provided for postmenopausal women below the lower tertile of a calcaneal broadband ultrasound attenuation (BUA) (< or = 58 db/MHz) or those with a SCORE value > or = 6. A telephone survey was conducted 6 mo after program registration to evaluate osteoporosis management outcomes. Of 271 women surveyed, 181 (67%) were postmenopausal and thus potential candidates for estrogen replacement, and 21% and 63% were assigned a high-risk profile by either calcaneal ultrasound or SCORE, respectively. Women at higher risk were significantly more likely to discuss osteoporosis with their family physician (p < 0.001), and to undergo further testing with dual X-ray absorptiometry measurement (p < 0.005). Utilization of estrogen replacement by postmenopausal women at time of survey was significantly increased compared to registration (p = 0.01). The self-referral program positively impacted decisions women made about preventing osteoporosis, particularly if they considered themselves at higher risk. PMID- 14742892 TI - Bone densitometry in premenopausal women: synthesis and review. AB - Bone loss prior to menopause is being increasingly identified in women. Clearly, low bone mineral density (BMD) is a significant risk factor for fracture in the estrogen-deficient female postmenopause. The significance of low bone density prior to menopause needs to be addressed. Low bone density in the premenopausal female may reflect attainment of a lower peak bone mass. It may also be secondary to progressive bone loss following achievement of peak bone density. The etiology of low bone density in the premenopausal female needs to be clarified with meticulous exclusion of secondary causes of bone loss. Menstrual status is an important determinant of peak bone mass as well as the development of bone loss in women prior to the onset of menopause. Subclinical decreases in circulating gonadal steroids may be associated with a lower peak bone mass as well as progressive bone loss in otherwise reproductively normal women. Elevations of follicle-stimulating hormone (FSH) of greater than 20 miu/L are associated with evidence of increased bone turnover marker activity and correlate with progressive bone loss in perimenopausal women. This transitional period requires further study with respect to the magnitude of bone loss experienced and the potential benefits of antiresorptive therapy. Detailed assessment of menstrual status is necessary in the evaluation of low bone density in premenopausal women. The majority of the cross-sectional and longitudinal studies completed evaluating BMD in the premenopausal years suggest that minimal bone loss does occur prior to menopause after attainment of peak bone mass. The magnitude of premenopausal bone loss, however, is controversial and may be site-dependent. More rapid rates of bone loss are seen in the transitional period beginning 2-3 yr prior to the onset of menopause. Prospective data are needed to understand further the relationship between BMD and fracture in the premenopausal period. Women with steroid-induced bone loss as well as other secondary causes of osteoporosis respond to antiresorptive therapy with documented improvements in BMD. Biomarkers can identify perimenopausal women with increased bone turnover. Lifestyle modification can improve BMD in the pre- and the perimenopausal period. Antiresorptive therapy has not been evaluated in pre- or perimenopausal women with low BMD in the absence of secondary causes of osteoporosis. As new treatment options are evaluated and become available, biomarker assessment may be of value in identifying women at risk of fracture. PMID- 14742893 TI - Seasonal variation in bone density in postmenopausal women. AB - We investigated seasonal variation in bone density by cross-sectional designs in two large cohorts of women (N = 2337) in Sweden. One cohort was strictly population-based (all 75 yr old, N = 1044), and one cohort was patient-based (age > or = 55 yr, mean age 68, N = 1293). Each woman was assessed once and the inclusion was continuous throughout almost all days of the year. Bone mineral density (BMD) of the total body, hip, and lumbar spine was determined by means of dual X-ray absorptiometry (DXA). For both cohorts, the BMD was similar in those assessed during the period April-September compared to those assessed during the period October-March. On the other hand, when women who had been assessed during July-December were compared with women assessed during January-June, we found the BMD to be up to 5% lower during July-December in all skeletal regions in the population-based sample, and 4% lower at the Ward's triangle and trochanter in the patient-based sample. After correction for small differences in age and weight, women of both cohorts who had been assessed during July-December still had lower BMD at the hip compared with women assessed during January-June. In conclusion, we found no variation in bone density during the period April September compared to the period October-March in these two cohorts of postmenopausal women. In elderly women living at a northern latitude, BMD may be a few percent lower during the period July-December compared to the period January-June. PMID- 14742894 TI - Quantitative ultrasound of the calcaneus as a screening tool to detect osteoporosis: different reference ranges for caucasian women, african american women, and caucasian men. AB - The interpretation of results measured by quantitative ultrasound (QUS) of the heel depends on the population studied. We measured estimated bone mineral density (BMD) of the heel using the Hologic Sahara sonometer. People were studied at county fairs, health fairs, and churches. Subjects were not on treatments that would affect bone density, other than calcium supplementation. This included 823 Caucasian women, 131 African American women, and 301 Caucasian men. In contrast to women, for Caucasian men the squared term for age was not significant, and a straight line of decline was the best fit for estimated BMD. African American women had a standard deviation larger than that reported by Hologic for Caucasian women. We compared a history of self-reported fractures with a subject's estimated BMD. An estimated BMD of 0.57 gm/cm2 included 75% of all fractures. This cutoff point was associated with increased fracture prevalence in subjects over age 50, relative risk of 1.4. This result corresponds to the Hologic data T score of -0.2. When used as a screening tool for osteoporosis fracture risk, an estimated BMD of 0.57 gm/cm2 seems reasonable in those subjects over age 50. PMID- 14742895 TI - Ability of peripheral DXA measurement to diagnose osteoporosis as assessed by central DXA measurement. AB - In order to evaluate the utility of peripheral measurement of bone mineral density (BMD) in the diagnosis of osteoporosis, we measured BMD at the spine and femoral neck with central dual-energy X-ray absorptiometry (DXA), at phalanx with AccuDXA (Schick) as well as proximal and distal forearm with pDXA (Norland) in 835 women ranging in age from 20 to 85 yr. In receiver operating characteristic (ROC) curves, where a positive case was defined as a T-score < or = -2.5 either on spine or femoral neck, the areas under the curve were not significantly different between sites. At a T-score of -2.5 as determined by each peripheral apparatus, sensitivity and specificity were, respectively, 0.39 and 0.95 for phalanx and 0.75 and 0.85 for proximal forearm whereas they were 0.42 and 0.96 for distal forearm. Using optimal absolute BMD cutoff values improved the results. Sensitivity and specificity were, respectively, 0.79 and 0.83 for phalanx at an absolute BMD value of 0.436 and 0.84 and 0.79 for proximal forearm at a value of 0.703, whereas they were 0.90 and 0.75 for distal forearm at a value of 0.208. Combining the two forearm measurements improves the results slightly. At cutoff values of 0.641 and 0.252, respectively for proximal and distal forearms, sensitivity was 0.83 and specificity was 0.84. Therefore, a peripheral measurement of BMD together with a good clinical evaluation of the osteoporosis risk profile of the patient, can be an interesting tool for the diagnosis of osteoporosis in areas where central DXA is not available. PMID- 14742896 TI - Selenium concentrations and glutathione peroxidase activities in blood of patients before and after allogenic kidney transplantation. AB - In animals and humans, the highest level of selenium (Se) occurs in the kidney. This organ is also the major site of the synthesis of the selenoenzyme glutathione peroxidase (GSH-Px). Decreased Se levels and GSH-Px activities in blood are common symptoms in the advanced stage of chronic renal failure (CRF). Blood samples for Se levels and GSH-Px activities measurements from patients were collected just before transplantation and 3, 7, 14, 30, and 90 d posttransplant. The Se levels in whole blood and plasma of patients before transplantation (79.5 and 64.5 ng/mL, respectively) were lower by 23% and 21%, respectively, as compared with controls (p < 0.0001), and 7 d after operation, it further decreased in both components (p < 0.01). Fourteen days after surgery, the levels reached the initial values and increased slowly in the later period. Red blood cell GSHPx activity in patients in the entire period of the study did not differ from the control group. Plasma GSH-Px of patients before the surgery was extremely low (76 U/L) as compared with controls (243 U/L; p < 0.0001) but increased rapidly to 115 U/L after 3 d, to 164 U/L after 14 d, and to 208 U/L after 3 mo posttransplant. In CRF patients, after kidney transplantation, plasma GSH-Px activity increased rapidly, approaching, after 3 mo, the values that were close to the normal levels. A negative correlation between creatinine level and plasma GSH-Px activity is observed in patients after kidney transplantation. Monitoring of plasma GSH-Px activity may be a useful additional marker of the transplanted kidney function. PMID- 14742897 TI - Selenium supplementation on plasma glutathione peroxidase activity in patients with end-stage chronic renal failure. AB - Patients with chronic renal failure (CRF) usually have a lower than healthy level of selenium (Se) in whole blood and plasma. Plasma glutathione peroxidase (GSH Px) is synthesized mostly in the kidney. In CRF patients, activity of this enzyme is significantly reduced and its reduction increases with the progress of the disease. The aim of the study was to evaluate the effect of Se supplementation to CRF patients at various stages of the disease on Se concentration in blood components and on plasma GSHPx activity. The study group comprised 53 CRF patients at various stages of the disease supplemented with Se (200 microg/d for 3 mo as Se-enriched yeast, containing about 70% L-selenomethionine [SeMet]). The control group consisted of 20 healthy subjects. The Se concentration in blood components was measured spectrofluorometrically with 2,3-diaminonaphthalene as a complexing reagent. GSH-Px activity in red cell hemolysates and plasma was assayed by the coupled method with tert-butyl hydroperoxide as a substrate. The Se concentration in whole blood and plasma of CRF patients is significantly lower as compared with healthy subjects, but similar at all stages of the disease. In the patients' plasma, total protein and albumin levels are also significantly lower than in healthy subjects. Plasma GSH-Px activity in patients is extremely low, and contrary to Se concentration, it decreases linearly with the increasing stage of the illness. Se-supplied patients show an increased Se concentration in all blood components and at all disease stages, whereas plasma GSH-Px activity is enhanced only at the incipient stage of the disease. Se supply has no effect on plasma GSHPx activity in uremic patients at the end stage of the disease. Total plasma protein and albumin levels did not change after Se supplementation. Our data seem to show that in patients with CRF lower total protein and albumin levels in plasma may be the chief cause of the low blood and plasma Se concentrations. GSH-Px activity decreases along with the kidney impairment. At the end stage of the disease, Se supplementation in the form of Se-enriched yeast has no effect on the increase in plasma GSH-Px activity. PMID- 14742898 TI - Alterations in plasma essential trace elements selenium, manganese, zinc, copper, and iron concentrations and the possible role of these elements on oxidative status in patients with childhood asthma. AB - The aim of the present study is to evaluate the status of plasma essential trace element selenium (Se), manganese (Mn), copper (Cu), zinc (Zn), and iron (Fe) concentrations and the effect of these elements on oxidative status in patients with childhood asthma. Plasma Se, Mn, Cu, and Zn concentrations were determined by atomic absorption spectrophotometry (AAS) and Fe concentrations, malondialdehyde (MDA), and total antioxidant capacity (TAC) were determined by the colorimetric method. The plasma MDA/TAC ratio was calculated as an index of oxidative status. Plasma albumin levels were measured to determine nutritional status. Plasma Fe concentrations, MDA levels and the MDA/TAC ratio were significantly higher (p<0.001, p<0.001, and p<0.01, respectively) and Se and Mn concentrations and TAC were lower (p<0.01, p<0.05, and p<0.01, respectively) in patients when compared to the healthy subjects. Plasma Zn, Cu, and albumin levels were not found to be significantly different in patients and controls (p>0.05). There were positive relationships between plasma MDA and Fe (r=0.545, p<0.001) and TAC and Se (r= 0.485, p<0.021), and a negative correlation between TAC and MDA values (r= -0.337, p<0.031) in patients with childhood asthma. However, there was no correlation between these trace elements and albumin content in patient groups. These observations suggest that increased Fe and decreased Se concentrations in patients with childhood asthma may be responsible for the oxidant/antioxidant imbalance. PMID- 14742899 TI - Maternal selenium deficiency increases hydrogen peroxide and total lipid peroxides in porcine fetal liver. AB - To investigate the role of selenium (Se) in the developing porcine fetus, prepubertal gilts (n = 42) were randomly assigned to either Se-adequate (0.39 ppm Se) or Se-deficient (0.05 ppm Se) gestation diets 6 wk prior to breeding. Maternal and fetal liver was collected at d 30, 45, 70, 90, and 114 of pregnancy. Concentrations of Se in maternal liver decreased during gestation in gilts fed the low-Se diet. The activity of cellular glutathione peroxidase (GPx) was decreased at d 30 and 45 of gestation in liver of gilts fed the low-Se diet. Concentrations of malondialdehyde (MDA) and hydrogen peroxide (H2O2) were greater in liver homogenates from gilts fed the low-Se diet. Within the fetuses, liver Se decreased in those fetuses of gilts fed the low-Se diet. Although the activity of GPx in fetal liver was not affected by the maternal diet, concentrations of H2O2 and MDA in fetal liver were greater in fetuses from gilts fed the low-Se diet. Maternal liver GPx activity was approx 12-fold greater than fetal liver GPx activity regardless of dietary treatment. These results indicate that maternal dietary Se intake affects fetal liver Se concentration and feeding a low-Se diet during gestation increases oxidative stress to the fetus, as measured by fetal liver H2O2 and MDA. PMID- 14742900 TI - Gestational changes in concentrations of selenium and zinc in the porcine fetus and the effects of maternal intake of selenium. AB - The effect of maternal dietary selenium (Se) and gestation on the concentrations of Se and zinc (Zn) in the porcine fetus were determined. Mature gilts were randomly assigned to treatments of either adequate (0.39 ppm Se) or low (0.05 ppm Se) dietary Se. Gilts were bred and fetuses were collected throughout gestation. Concentrations of Se in maternal whole blood and liver decreased during gestation in sows fed the low-Se diet compared to sows fed the Se-supplemented diet. Maternal intake of Se did not affect the concentration of Se in the whole fetus; however, the concentration of Se in fetal liver was decreased in fetuses of sows fed the low-Se diet. Although fetal liver Se decreased in both treatments as gestation progressed, the decrease was greater in liver of fetuses from sows fed the low- Se diet. Dietary Se did not affect concentrations of Zn in maternal whole blood or liver or in the whole fetus and fetal liver. The concentration of Se in fetal liver was lower but the concentration of Zn was greater than in maternal liver when sows were fed the adequate Se diet. These results indicate that maternal intake of Se affects fetal liver Se and newborn piglets have lower liver Se concentrations compared to their dams, regardless of the Se intake of sows during gestation. Thus, the piglet is more susceptible Se deficiency than the sow. PMID- 14742901 TI - Effect of selenium on lipids, lipid peroxidation, and sulfhydryl group in neuroendocrine centers of rats. AB - The effects of various doses of sodium selenite (0.05, 0.1, and 0.2 mg/kg body weight, i.p.) were studied on the content of phospholipids, cholesterol, esterified fatty acids (EFA), gangliosides, thiobarbituric acid reactive substance (TBARS), and sulfhydryl group in neuroendocrine centers of male Wistar rats for 7 d. The lowest dose of Se (0.05 mg/kg) did not alter the above parameters significantly in neuroendocrine centers. The content of phospholipids was depleted significantly in the pituitary and depletion in the pineal was 80.22% with a 0.1-mg/kg dose of Se, but this dose elevated its level significantly in the hypothalamus. Conversely, a 0.2-mg/kg dose of selenium elevated the level of phospholipids significantly in the pituitary and hypothalamus, the elevation in the pineal was 70%. Selenium, 0.1 mg/kg, elevated the level of cholesterol in the pituitary but depleted its level in the pineal (56.8%) and hypothalamus (13.60%). Selenium, 0.2 mg/kg, elevated the level of cholesterol significantly in the hypothalamus but its level was not significant in the pituitary and pineal. The depletion of esterified fatty acid in the pituitary and pineal with doses of 0.1 and 0.2 mg/kg was significant in the pituitary, whereas its depletion in the pineal was 85.4% and 69.26%, respectively. Selenium, 0.1 and 0.2 mg/kg, depleted the level of gangliosides significantly and dose dependently in the pituitary but has elevated its level significantly and dose dependently in the hypothalamus. Its depletion in the pineal was 87.1% and 67.8% with the 0.1- and 0.2-mg/kg dose of selenium, respectively. Selenium, 0.1 mg/kg, increased the content of TBARS significantly in neuroendocrine centers and its elevation in the pineal was 703.8%. Selenium, 0.2 mg/kg, elevated its level in the pituitary and it was 126.9% in the pineal, but this dose depleted its level significantly in the hypothalamus. The content of the sulfhydryl group with a 0.1-mg/kg dose of selenite was depleted significantly in neuroendocrine centers and it was 55.9% in the pineal. Selenium, 0.2 mg/kg, depleted the level of the sulfhydryl group more significantly in the pituitary and pineal, but its elevation in hypothalamus was significant. PMID- 14742902 TI - Effect of selenium-induced oxidative stress on the oxidation reduction system and reproductive ability of male mice. AB - The present study was carried out to evaluate the effect of selenium (Se)-induced oxidative stress on the oxidation reduction system and the fertility status of male mice. Different levels of Se, a potent antioxidant, were fed in three separate groups for 8 wk to create the different oxidative stress in mice. A significant decrease in the glutathione peroxidase (GSH-Px) in both liver and testis was observed in the Se-deficient (0.02 ppm) group I, whereas enzyme levels in the Se-excess (1 ppm) group were comparable to the Se-adequate (0.2 ppm) group. Glutathione-S-transferase activity was enhanced in group I in comparison to group II; however, no change was seen in group III. The glutathione reductase and superoxide dismutase activities were decreased in the Se-deficient group, whereas the enzyme levels were significantly increased in the Se-excess group. The fertility status of the animals studied in terms of percentage fertility and litter size showed a significant decrease in the reproductive ability of male mice in group I when compared to group II. No changes in the fertility status of animals were observed in group III. Thus, the data clearly indicate the effect of oxidative stress generated by feeding various Se levels on the oxidation reduction system and, consequently, its effect on the reproductive ability of male mice. PMID- 14742903 TI - Effect of selenium depletion and supplementation on the kinetics of type I 5' iodothyronine deiodinase and T3/T4 in rats. AB - Presently, the effect of selenium (Se) deficiency and excess of Se (1 ppm) on the activity of selenoenzymes type 1 5'-iodothyronine deiodinase (5'-DI), glutathione peroxidase (GSH-Px), and level of thyroid hormones (T3 and T4) was studied in rats. Se levels in the serum and liver, T3 and T4 in the serum, GSH-Px levels in the liver, and 5'-DI activity in the liver/aorta/thyroid were estimated after 1, 2, and 3 mo of Se-deficient (0.02 ppm), Se-adequate (0.2 ppm), and Se-excess (1 ppm) diet feeding. All of these parameters decreased significantly in the Se deficient group as compared to the adequate group. Within the deficient group, as the Se deficiency progressed, all of the parameters except 5'-DI decreased after 2 and 3 mo in comparison to 1-mo data. Thyroidal 5'-DI activity in Se deficiency showed the maximum increase. A significant increase was observed in all of the above parameters in the 1 ppm Se-supplemented diet group when compared with the adequate Se group; also, as the Se deposition increased within the Se-excess diet group, a significant increase was observed in all of the above parameters. However, as observed by others, the intake of excess of Se (i.e., 2 ppm in the diet) did not elevate the activities of selenoenzymes and thyroid hormones; rather, it had adverse effects. The present study concludes that Se supplementation at least up to 1 ppm enhances the selenoenzyme activities, and above this level, it may not be considered as an indicator of selenoenzyme activities. PMID- 14742904 TI - Proceedings of the 2003 Summer Neuropeptide Conference, Montauk, New York. PMID- 14742905 TI - Neprilysin regulates amyloid Beta peptide levels. AB - That neprilysin (NEP) is a major Abeta peptide-degrading enzyme in vivo is shown by higher Abeta peptide levels in the brain of an NEP knockout mouse. In addition, we show that infusion of an NEPinhibitor, but not inhibitors of other peptidases, into the brains of an APP transgenic mouse elevates Abeta levels. We have investigated the use of NEP as a potential therapeutic agent to prevent the accumulation of Abeta peptides in the brain. Lentivirus expressing NEP was initially used to demonstrate the ability of the enzyme to reduce Abeta levels in a model CHO cell line and to make primary hippocampal neurons resistant to Abeta mediated neurotoxicity. Injection of NEPexpressing lentivirus, but not inactive NEP-expressing lentivirus, GFP-expressing lentivirus, or vehicle, into the hippocampus of 12-20-mo-old hAPP transgenic mice led to an approx 50% reduction in the number of amyloid plaques. These studies provide the impetus for further investigating of the use of NEP in a gene transfer therapy paradigm to prevent the accumulation of Abeta and prevent or delay the onset of Alzheimer's disease. PMID- 14742906 TI - Overexpression of human cystatin C in transgenic mice does not affect levels of endogenous brain amyloid Beta Peptide. AB - Cystatin C, an inhibitor of cysteine proteases, colocalizes with amyloid beta (Abeta) in parenchymal and vascular amyloid deposits in brains of Alzheimer's disease (AD) patients, suggesting that cystatin C has a role in AD. Cystatin C also colocalizes with beta amyloid precursor protein (betaAPP) in transfected cultured cells. In vitro analysis of the association between the two proteins revealed that binding of cystatin C to full-length betaAPP does not affect the level of Abeta secretion. Here we studied the effect of in vivo overexpression of cystatin C on the levels of endogenous brain Abeta. We have generated lines of transgenic mice expressing either wild-type human cystatin C or the Leu68Gln variant that forms amyloid deposits in the cerebral vessels of Icelandic patients with hereditary cerebral hemorrhage, under control sequences of the human cystatin C gene. Western blot analysis of brain homogenates was used to select lines of mice expressing various levels of the transgene. Analysis of Abeta40 and Abeta42 concentrations in the brain showed no difference between transgenic mice and their nontransgenic littermates. Thus, in vivo overexpression of human cystatin C does not affect Abeta levels in mice that do not deposit Abeta. PMID- 14742907 TI - First evidence for helical transitions in supercoiled DNA by amyloid Beta Peptide (1-42) and aluminum: a new insight in understanding Alzheimer's disease. AB - Previously, we evidenced a B --> Z helical change in Alzheimer's brain genomic DNA, leading to a hypothesis that Alzheimer's disease (AD) etiological factors such as aluminum (Al), amyloid beta (Abeta) peptide, and Tau might play a role in modulating DNA topology. In the present study, we investigated the interaction of Al and Abeta with DNA. Our results show that Abeta(1-42) could induce a B --> Psi (Psi) conformational change in pUC 18 supercoiled DNA (scDNA), Abeta(1-16) caused an altered B-form, whereas Al induced a complex B-C-A mixed conformation. Ethidium bromide binding and agarose gel electrophoresis studies revealed that Al uncoiled the DNAto a fully relaxed form, whereas Abeta(1-42) and Abeta(1-16) effected a partial uncoiling and also showed differential sensitivity toward chloroquine-induced topoisomer separation. Our findings show for the first time that Abeta and Al modulate both helicity and superhelicity in scDNA. A new hypothetical model explaining the potential toxicity of Abeta and Al in terms of their DNA binding properties leading to DNA conformational alteration is proposed. PMID- 14742908 TI - NCAM mimetic peptides: Pharmacological and therapeutic potential. AB - The neural cell adhesion molecule (NCAM) plays an important role in neuronal differentiation and synaptic plasticity, making it an attractive target for the development of drugs for the treatment of neurodegenerative disorders. NCAM binds to itself (homophilic binding) and to a series of counter-receptors, including the fibroblast growth factor receptor (FGFR), other adhesion molecules, and various extracellular matrix components (heterophilic binding). By means of combinatorial chemistry and based on the unraveling of the structure of NCAM, it has been possible to develop a number of peptides that mimic NCAM homophilic binding. These peptides interfere with cell adhesion and promote differentiation and cell survival. Recently, a peptide mimicking the heterophilic binding to FGFR has also been identified. It binds and activates the receptor, thereby modulating neurite extension and synaptic plasticity. PMID- 14742909 TI - Is there structural specificity in the reversible protein aggregates that are stored in secretory granules? AB - There are several steps that must occur for secretory granules to form: (1) Secretory proteins that make up the dense cores of the granules must be concentrated; (2) membrane proteins necessary for granule function must accumulate in the correct location; and (3) inappropriate membrane proteins and excess membrane must be removed. Reversible aggregation of secretory granule proteins provides a mechanism for concentrating and sorting these proteins. There is specificity in the way secretory granule proteins are treated in cells that make granules. The specificity has been shown in some cases to occur after the aggregation process, so that granules containing different aggregates function differently. An explanation could be that a property of the aggregate, such as a surface motif, might influence the accumulation of membrane proteins necessary for granule function. Such a conclusion implies that the aggregates are not amorphous but have structure. Use of NMR spectroscopy to investigate changes in the environment of amino acid residues in secretory granule proteins as they form oligomers by using 15N relaxation times might provide a means to determine which residues are specifically involved in aggregation. PMID- 14742910 TI - Kalirin expression is regulated by multiple promoters. AB - Kalirin is a multidomain guanine nucleotide exchange factor for small GTPbinding proteins of the Rho family. It is expressed in multiple isoforms that contain different combinations of functional domains and display a complex pattern of expression during brain development. In addition to the isoforms generated through alternative splicing, we have identified multiple transcriptional start sites in rats and humans. These multiple transcriptional start sites result in full-length Kalirin transcripts possessing different 5' ends encoding proteins with differing amino termini. These alternative first exons display different patterns of expression in developing rats and humans and in cultured cells. Most of these alternate first exons lie >100 kb upstream of exon 2 in both rats and humans. Comparisons of the rat and human Kalirin promoter regions reveal numerous shared potential regulatory elements. PMID- 14742911 TI - Secretory granule biogenesis and neuropeptide sorting to the regulated secretory pathway in neuroendocrine cells. AB - Neuropeptide precursors synthesized at the rough endoplasmic reticulum are transported and sorted at the trans-Golgi network (TGN) to the granules of the regulated secretory pathway (RSP) of neuroendocrine cells. They are then processed into active peptides and stored in large dense-core granules (LDCGs) until secreted upon stimulation. We have studied the regulation of biogenesis of the LDCGs and the mechanism by which neuropeptide precursors, such as pro opiomelanocortin (POMC), are sorted into these LDCGs of the RSP in neuroendocrine and endocrine cells. We provide evidence that chromogranin A (CgA), one of the most abundant acidic glycoproteins ubiquitously present in neuroendocrine/endocrine cells, plays an important role in the regulation of LDCG biogenesis. Specific depletion of CgA expression by antisense RNAs in PC12 cells led to a profound loss of secretory granule formation. Exogenously expressed POMC was neither stored nor secreted in a regulated manner in these CgA-deficient PC12 cells. Overexpression of CgA in a CgA- and LDCG-deficient endocrine cell line, 6T3, restored regulated secretion of transfected POMC and the presence of immunoreactive CgA at the tips of the processes of these cells. Unlike CgA, CgB, another granin protein, could not substitute for the role of CgA in regulating LDCG biogenesis. Thus, we conclude that CgA is a key player in the regulation of the biogenesis of LDCGs in neuroendocrine cells. To examine the mechanism of sorting POMC to the LDCGs, we carried out site-directed mutagenesis, transfected the POMC mutants into PC12 cells, and assayed for regulated secretion. Our previous molecular modeling studies predicted a three-dimensional sorting motif in POMC that can bind to a sorting receptor, membrane carboxypeptidase E (CPE). The sorting signal consists of four conserved residues at the N-terminal loop structure of POMC: two acidic residues and two hydrophobic residues. The two acidic residues were predicted to bind to a domain on CPE (CPE254-273) containing two basic residues (R255 and K260) to effect sorting into immature secretory granules. Site-directed mutagenesis of the motif on POMC resulted in accumulation of the mutant in the Golgi, as well as high basal secretion, indicating that the mutant POMC was inefficiently sorted to the RSP. These results support the model that POMC is actively sorted to the RSP granules for processing and secretion by a sorting signal-mediated mechanism. PMID- 14742912 TI - Transcriptional activation of the human gastrin-releasing peptide receptor gene in gastrointestinal and prostatic epithelial cancer cells. AB - The mammalian gastrin-releasing peptide receptor (GRP-R) belongs to the superfamily of G protein-coupled receptors and mediates actions of the regulatory GRP and bombesin, the amphibian homolog of GRP. Owing to its frequent ectopic expression in some epithelial human malignancies, such as cancers of the colon, lung, and prostate, ligand-specific receptor activation may initiate intracellular signals of cell proliferation, differentiation and migration in this context. Because the underlying molecular mechanisms of aberrant human GRP-R (hGRP-R) expression in tumorigenesis remain unknown, we examined in this study the transcriptional activation of hGRP-R in gastrointestinal and prostate cancer cells, which natively express functional hGRP-R. Using various hGRP-R promoter mutants cloned into a luciferase reporter plasmid, transient transfection studies demonstrated robust transcriptional activation in gastrointestinal and prostate cancer cells. Although our study revealed distinct patterns of transcriptional hGRP-R activation in gastrointestinal and prostate cancer cells, genomic sequences between 97 and 247 bp upstream of the major RNA initiation site appear to be of particular significance for basal transcriptional hGRP-R activation. Based on this study, future examination of transcription factor interaction with the hGRP-R promoter will be important to identify molecular mechanisms of hGRP-R regulation relevant in human cancers that express functional receptor sites PMID- 14742913 TI - Differential coupling of the PAC1 SV1 splice variant on human colonic tumors to the activation of intracellular cAMP but not intracellular Ca2+ does not activate tumor proliferation. AB - PAC1 is a recently cloned and characterized heptahelical, G protein-coupled receptor with high affinity to PACAP-27 and PACAP-38 and is differentially coupled to activate intracellular Ca2+ and cAMP. PAC1 is expressed as four major splice variants, each possessing differential coupling to inositol phosphates and intracellular Ca2+. PAC1 has been shown previously to be expressed and regulate the growth and proliferation of nonsquamous cell lung cancer cells, as well as breast cancer cell lines. PAC1 is expressed on the HCT8 human colon cancer cell line and is coupled to the activation of both intracellular cAMP and Ca2+ with consequent stimulation of growth. In the current study, we contrast the effects of pituitary adenylate cyclase-activating polypeptide (PACAP) on the HCT8 colon cancer cell lines to the HCT116 and FET cell lines wherein PAC1 is expressed as the SV1 or HIP splice variant and is coupled to the activation only of cAMP but not of intracellular Ca2+. These data indicate that human colon tumor cells express PAC1 and are differentially coupled to intracellular signal transduction molecules. The ability to activate both cAMP and Ca2+ appears to be a prerequisite for activation of tumor proliferation, indicating a potentially important factor in how PACAP potentiates the growth of certain tumors. PMID- 14742914 TI - The prolactin-releasing peptide receptor (GPR10) regulates body weight homeostasis in mice. AB - To identify new drug targets for the treatment of obesity, we employed a degenerate reverse transcriptasepolymerase chain reaction technique to isolate novel members of the G-protein coupled receptor superfamily from mouse hypothalamus. One of our clones was found to encode a protein with 90% amino acid identity to human GPR10, which was previously identified as the receptor for prolactin-releasing peptide (PrRP) and has been implicated in lactation, the regulation of food intake and other physiological functions. To investigate the role of GPR10 in food intake and body weight homeostasis, we generated mice carrying a targeted deletion of the GPR10 gene. First, using these knockout animals, we confirmed that GPR10 is the principle receptor for PrRP in the mouse hypothalamus because deletion of GPR10 completely abolished PrRP binding to isolated hypothalamic cell membranes. Second, we investigated the effect of normal and high-fat diets on energy intake, body weight, and glucose homeostasis in wild-type and GPR10 knockout mice. After fasting and refeeding, food intake in knockout animals was unchanged relative to control littermates. However, beginning at 16 wk of age on a normal diet, knockout mice became hyperphagic, obese, and showed significant increases in body fat and the levels of leptin and insulin, as well as decreased glucose tolerance. This metabolic profile was similar to the effect of a high-fat diet on wild-type animals. Our findings provide direct evidence that GPR10 is the receptor for PrRP and that it is involved in the regulation of energy balance in mice. GPR10 knockout mice will also prove useful for investigating other proposed activities for PrRP. PMID- 14742915 TI - The pharmacology of adrenomedullin receptors and their relationship to CGRP receptors. AB - Adrenomedullin (AM) has two specific receptors formed by the calcitonin-receptor like receptor (CL) and receptor activity-modifying protein (RAMP) 2 or 3. These are known as AM1 and AM2 receptors, respectively. In addition, AM has appreciable affinity for the CGRP1 receptor, composed of CL and RAMP1. The AM1 receptor has a high degree of selectivity for AM over CGRP and other peptides, and AM22-52 is an effective antagonist at this receptor. By contrast, the AM2 receptor shows less specificity for AM, having appreciable affinity for betaCGRP. Here, CGRP8-37 is either equipotent or more effective as an antagonist than AM22-52, depending on the species from which the receptor components are derived. Thus, under the appropriate circumstances it seems that betaCGRP might be able to activate both CGRP1 and AM2 receptors and AM could activate both AM1 and AM2 receptors as well as CGRP1 receptors. Current peptide antagonists are not sufficiently selective to discriminate between these three receptors. The CGRP-selectivity of RAMP1 and RAMP3 may be conferred by a putative disulfide bond from the N-terminus to the middle of the extracellular domain of these molecules. This is not present in RAMP2. PMID- 14742916 TI - The assessment of vasoactive properties of CGRP and adrenomedullin in the microvasculature: a study using in vivo and in vitro assays in the mouse. AB - The potent neuropeptide vasodilator, calcitonin gene-related peptide (CGRP), and the vasoactive peptide adrenomedullin (AM) are structurally related. Evidence from our laboratory has demonstrated that these peptides have potent microvascular actions of relevance to cardiovascular and inflammatory effects in health and disease. We wish to further investigate the actions of these peptides through studies in genetically modified mice. We have developed techniques to enable the quantitative analysis of CGRP and AM responses in the mouse microvasculature. A mouse isolated mesentery system was developed that measures changes in perfusion pressure used as an index of microvascular relaxation in the precontracted mesenteric microvascular bed. Bolus injections of CGRP and AM caused dose-dependent decreases in perfusion pressure that were proportional to vascular relaxation. An in vivo mouse skin assay was also used in which agents were injected intradermally into the dorsal skin. The effects of these agents was assessed by the extravascular accumulation of intravenously injected 125I-albumin for their ability to potentiate plasma extravasation induced by a mediator of increased microvascular permeability. CGRP and AM are not directly active in this assay, because it does not directly measure blood flow. However, the vasodilators acted in a potent and dose-dependent manner to significantly potentiate edema formation. The results demonstrate the potent activity of CGRP and the activity (although 100- to 300-fold less potent) of AM. Furthermore, the results demonstrate the increased potency of CGRP in the microvasculature when compared with the structurally distinct peptide VIP and PGE1. PMID- 14742917 TI - Attenuation of knee joint inflammation by peripherally administered endomorphin 1. AB - Endomorphin-1 is a selective endogenous ligand for the micro-opioid receptor, and this study investigated the effect of endomorphin-1 on rat knee joint inflammation by examining the ability of the neuropeptide to modulate synovial protein extravasation. Acute joint inflammation was induced by intraarticular injection of 2% kaolin followed by 2% carrageenan and the animals allowed to recover for 3 h. Immunohistochemical examination of these inflamed joints revealed endomorphin-1-like immunoreactive nerves in deep synovium with a proportion of the nerve fibers occurring in close proximity to synovial blood vessels. Perfusion of inflamed knees with exogenous endomorphin-1 across the dose range 10-9-10-6 M produced a significant reduction in synovial vascular permeability with the 10-7M dose producing the greatest fall in protein exudation (approx 55%). These effects were blocked by the specific micro-opioid receptor antagonist CTOP. Destruction of knee joint unmyelinated afferent nerve fibers by capsaicin treatment significantly attenuated the anti-inflammatory effects of endomorphin-1, suggesting that the peptide is acting via a neurogenic mechanism. The findings of this study indicate that endomorphin-1 acts peripherally in knee joints to reduce synovial protein extravasation. These anti-inflammatory effects are mediated by micro-opioid receptors located on capsaicin-sensitive afferent nerves. PMID- 14742918 TI - Growth hormone localization in the neural retina and retinal pigmented epithelium of embryonic chicks. AB - It is well-established that growth hormone (GH) is present in the brain, spinal cord, and peripheral nerves of embryonic chicks, prior to the differentiation of pituitary somatotrophs, but its presence and distribution in retinal tissues is controversial. The possible presence of GH and GH mRNA in retinal tissues of early embryos has therefore been further evaluated. A 466-bp fragment of the pituitary GH cDNA, derived from a portion of exon 3 and spanning exons 4 and 5, was amplified by RT-PCR from reverse-transcribed mRNA from the pituitary glands of juvenile chicks and from the whole eye, neural retina, and retinal pigmented epithelium (RPE) of embryonic-day (ED) 9 chick embryos. In ED 9 embryos, GH immunoreactivity was demonstrated in the choroid and neural retina, in which it was particularly abundant in a layer of cells with the location and morphological appearance of retinal ganglion cells. GH immunoreactivity was also present in tissue sections of the RPE that were bleached to remove the melanin pigment. The intense GH staining in the RPE of ED 9 embryos was also revealed using a fluorescein-labeled GH antibody and confocal microscopy. At the ultrastructural level, GH detected by immunogold electron microscopy was present in the cytoplasm of RPE and neural retinal cells of ED 9 embryos. Although not associated with secretory granules, GH in the RPE was particularly associated with the membranes of the melanin granules. These results demonstrate that the neural retina and RPE are extrapituitary sites of GH production in early chick embryos, prior to the differentiation of the pituitary gland. PMID- 14742920 TI - Nonconvulsive status epilepticus: an under diagnosed and potentially treatable condition. PMID- 14742919 TI - Novel insect control agents based on neuropeptide antagonists: The PK/PBAN family as a case study. AB - This review describes the development of a new integrated approach to the generation of a novel type of insect neuropeptide (Np) antagonists and putative insect control agents based on conformationally constrained compounds. The new approach, termed insect Np-based antagonist insecticide (INAI), was applied to the insect pyrokinin (PK)/pheromone biosynthesis-activating Np (PBAN) family as a model and led to the discovery of a potent linear lead antagonist and several highly potent, metabolically stable backbone cyclic (BBC) conformationally constrained antagonists that were devoid of agonistic activity and inhibited sex pheromone biosynthesis in female moths in vivo. This review summarizes the above approach, briefly describes the PK/PBAN Np family, presents data on the in vivo activity of the antagonists, summarizes data on the PK/PBAN receptor, and introduces the advantages of this method for generation of Np antagonists as a basis for the design of insect control agents. PMID- 14742921 TI - Botulinum toxins: pharmacology and its current therapeutic evidence for use. AB - Botulinum toxins are, as a group, among the most potent neuromuscular toxins known, yet they are clinically useful in the management of conditions associated with muscular and glandular over-activity. Botulinum toxins act by preventing release of acetylcholine into the neuromuscular junction. While botulinum toxin type A is commonly available, different manufacturers produce specific products, which are not directly interchangeable and should not be considered as generically equivalent formulations. Type B is also available in the market. Each formulation of botulinum toxin is unique with distinct dosing, efficacy and safety profiles for each use to which it is applied. Botulinum toxin type A is the treatment of choice based on its depth of evidence in dystonias and most other conditions. Botulinum toxin type A is established as useful in the management of spasticity, tremors, headache prophylaxis and several other neurological conditions. Active research is underway to determine the parameters for which the type B toxin can be used in these conditions, as covered in this review. Botulinum toxin use has spread to several fields of medicine. PMID- 14742922 TI - The management of non-functioning pituitary adenomas. AB - Non-functioning pituitary adenomas most commonly present secondary to mass effect and are classified according to their size and immunohistochemical staining. Local intrasellar mass effect may cause varying degrees of hypopituitarism. With extrasellar growth, neurological signs and symptoms develop. Appropriate therapy for these tumors requires close interaction across multiple disciplines. Trans sphenoidal surgery offers safe and effective treatment in the overwhelming majority of patients with relatively low risk of new neurological and endocrinologic deficits. The multidisciplinary management of non-functioning adenomas, their diagnosis and therapeutic outcomes, is discussed. PMID- 14742923 TI - Nervous system involvement in asymptomatic HIV seropositive individuals: a cognitive and electrophysiological study. AB - BACKGROUND: A significant proportion of human immunodeficiency virus (HIV) seropositive subjects may have subclinical asymptomatic involvement of the central and peripheral nervous system. AIMS: To detect subclinical neurological involvement in asymptomatic HIV seropositive individuals by clinical, mini mental state examination (MMSE) and various electrophysiological tests. MATERIAL AND METHODS: MMSE, EEG, nerve conduction (NC), and multimodality-evoked potential studies were evaluated in 20 asymptomatic HIV seropositive subjects. RESULTS: The majority of the subjects were young (mean age: 29.5 +/- 8.9 yrs) and female (85%). The mean MMSE score was 25.8 +/- 2.3, which was marginally less than that of matched controls (26.3 +/- 2.4). Four subjects had a score of less than 23 suggesting subclinical cognitive impairment. EEG (n=19) was mildly abnormal in 8 cases: low alpha index (n=4), excess of fast background activity (n=3) and intermittent bursts of slow waves along with sometimes sharp waves (n=4). Motor NC studies (n=20) revealed a decreased mean nerve conduction velocity (NCV) compared to controls in the right median (P<0.05) and CP (P<0.001) nerves. Sensory NC studies revealed a decreased mean NCV in both the median (P<0.001) and sural (P<0.001) nerves compared with controls. Brainstem auditory evoked potential (BAEP) studies showed the involvement of the peripheral auditory pathway (23.5%). The somatosensory evoked potential (SSEP) study detected a delay of N20 latency in two. Only 3 subjects came for follow-up after 6 months. CONCLUSION: Asymptomatic HIV seropositive subjects may have subclinical central and peripheral nervous system involvement. Long-term follow-up studies are essential for better understanding of the significance of these changes. PMID- 14742924 TI - Feasibility of acute thrombolytic therapy for stroke. AB - BACKGROUND: Thrombolysis is an expensive medical intervention for ischemic stroke and hence there is a need to study the feasibility of thrombolysis in rural India. AIMS: To asses the feasibility and limitations of providing thrombolytic therapy to acute ischemic stroke patients in a rural Indian set-up. MATERIAL AND METHODS: The first 64 consecutive patients registered under the Acute Stroke Registry in a university referral hospital with a rural catchment area were studied as per a detailed protocol and questionnaire. RESULTS: Of the 64 patients 44 were ischemic strokes, and 20 were hemorrhagic. Thirteen (29.55%) patients with ischemic stroke reached a center with CT scan facility within 3 hours, of whom only 7 (15.91%) were eligible to receive thrombolytic therapy as per the existing clinical and radiological criteria, but none received the therapy. Of the remaining 31 (70.45%) who arrived late, 11 (25%) had no clinical and radiological contraindications for thrombolysis, except the time factor. All the patients belonged to a low socioeconomic status and a rural background. CONCLUSION: Though a large proportion of ischemic stroke patients were eligible to receive thrombolytic therapy, the majority could not reach a center with adequate facilities within the recommended time window. More alarmingly, even for those patients who reached within the time window, no significant attempt was made to initiate thrombolysis. These data call not only for attention to improve existing patient transport facilities, but also for improving the awareness of efficacy and therapeutic window of thrombolysis in stroke, among the public as well as primary care doctors. PMID- 14742925 TI - Role of in vivo proton MR spectroscopy in the evaluation of adult brain lesions: our preliminary experience. AB - CONTEXT: A definite diagnosis and characterization of intracranial mass lesions, based on structural Magnetic Resonance Imaging (MRI) alone may be difficult. In such cases Proton Magnetic Resonance Spectroscopy (1H-MRS) along with other non invasive techniques represents an advance in the specificity of brain lesion diagnosis. AIMS: The primary aim of this study was to evaluate the extent of the utility of 1H-MRS in adult brain tumors and their differentiation from similar appearing space-occupying lesions. MATERIAL AND METHODS: MRS studies were performed on 1.5 Tesla whole body MR system using standard imaging head coil. Sixty patients (aged 30-65 years), including 35 males (31-65 years) and 25 females (30-65 years) were studied, along with 25 age-matched healthy volunteers (30-64 years). The Student 't' test was used to statistically analyze the spectroscopic data for significant difference in the metabolite ratios of the lesions from normal brain tissue. RESULTS: The Cho/Cr ratio was significantly raised in low and high-grade glioma and meningioma patients (1.85 +/- 0.36, 3.50 +/- 1.00 and 6.65 +/- 2.83 respectively) (mean +/- standard deviation), as compared with the control group (1.16 +/- 0.18); and NAA/Cr and NAA/Cho ratios were found to be lower than normal values in our study (P<0.01). However, in the non-neoplastic lesions, the Cho/Cr ratios were not statistically significant. The tubercular lesions revealed an average Cho/Cr ratio of 1.24 +/- 0.18, while it was 1.14 +/- 0.07 for infarcts. CONCLUSION: MR Spectroscopy was useful to arrive at a more definitive diagnosis in doubtful intracranial space-occupying lesions with similar morphological imaging patterns. PMID- 14742927 TI - Experimental study of rapid versus slow sagittal sinus occlusion in dogs. AB - BACKGROUND: Clinical, radiological, postmortem and experimental studies are not enough for the definition of pathophysiological differences between rapid and slow-progressing cerebral venous system obstruction. AIMS: An experimental study was conducted to set some physiopathological differences between rapid and slow occlusion of the superior sagittal sinus. SETTINGS AND DESIGN: Eighteen dogs categorized into 3 groups were chosen as test subjects. The three groups were the rapid occlusion, slow occlusion and the control study groups and each group had six subjects. MATERIAL AND METHODS: Intracranial pressure values, histopathological findings, and the degree of cerebral edema formation, estimated by measuring the water content ratio of the brain and the angiographic results in the 2 different groups of subjects that underwent rapid and slow superior sagittal sinus obstruction were compared with that of the control subjects. STATISTICAL ANALYSIS: Statistical analysis was performed using GraphPad Prisma V.3 statistical software. Variables of the 3 groups were compared using non parametric Kruskal Wallis ANOVA test and multiple comparisons were made using Dunn's multiple test. The comparison of initial and terminal intracranial pressure values obtained before and after the sinus occlusion, was made using the Wilcoxon test. A probability value of less than 0.05 was regarded as significant. RESULTS AND CONCLUSIONS: Comparison of the water content ratio of the brain in the 3 groups, the difference between the initial and terminal intracranial pressure values of the rapid occlusion study group, and the difference between the terminal intracranial pressure values of the 3 groups was statistically significant (P<0.05). Dunn's Multiple Comparison Test yielded significant differences in the water content ratio of the brain and in the intracranial pressure values between the rapid occlusion study group and the control group (P<0.05). Moreover, histopathological and radiological examination disclosed more prominent brain edema findings, and less apparent collateral venous flow in the rapid occlusion study group than in the slow occlusion one. To conclude, the clinical severity of sinus occlusion seems directly related to the quickness of the occlusion and the capacity of the collateral venous system. PMID- 14742926 TI - The effect of cranioplasty on cerebral hemodynamics: evaluation with transcranial Doppler sonography. AB - BACKGROUND: The purpose of cranioplasty is not only cosmetic repair but also neurological improvement. The effect of cranioplasty on the cerebral hemodynamics flow has not been investigated by ultrasonographic techniques. AIMS: To investigate changes of cerebral hemodynamics after cranioplasty in patients with cranial defect using transcranial Doppler sonography (TCDS). SETTING: The Departments of Neurosurgery and Radiology of a university hospital. DESIGN: A prospective clinical study. MATERIAL AND METHODS: We prospectively examined the cerebral hemodynamics with TCDS pre- and postoperatively in 18 patients with cranial defect who underwent cranioplasty. All postoperative studies were done between the 7th and 15th day after cranioplasty. The anterior cerebral artery was examined through the transtemporal and transorbital windows, the middle cerebral artery through the transtemporal window, and the posterior cerebral artery through the transforaminal window. Bilaterally, the peak systolic, end diastolic and mean blood flow velocities of these arteries were measured. STATISTICAL ANALYSIS: Wilcoxon matched-pairs signed-ranks test. RESULTS: Before cranioplasty all the velocities ipsilateral to the cranial defect were significantly low, while in the contralateral side they were near normal. Ipsilateral low cerebral blood flows increased and reached normal levels (P<0.05) after cranioplasty. During the follow-up, neurological improvement was observed. CONCLUSION: Cranioplasty is carried out not only for preserving normal appearances and physical barrier but also for neurological improvement. This should be explained by the normalization of cerebral hemodynamics. PMID- 14742928 TI - Intravenous immunoglobulin reduces serum tumor necrosis factor alpha in patients with Guillain-Barre syndrome. AB - BACKGROUND: Tumor necrosis factor a TNF-alpha has a possible role in the pathogenesis of the Guillain-Barre syndrome (GBS). AIMS: To study the effect of intravenous immunoglobulin (IVIg) on serum TNF-alpha concentrations in patients with GBS. MATERIAL AND METHODS: The effect of IVIg on TNF-alpha was evaluated in 36 patients with GBS. Serum TNF-alpha concentration was measured by enzyme-linked immunosorbent assay (ELISA). The sera of 22 (61%) patients with GBS showed elevated concentrations of TNF-alpha (35-182 pg/ml) and these sera were individually incubated in vitro with IVIg (0.25 mg/ml) at 37 degrees C for 24 hours. RESULTS: The serum TNF-alpha concentrations in the 22 GBS patients with elevated levels showed a steady decline (60.34-19.78 pg/ml) following incubation with IVIg. These 22 patients also received IVIg therapy, and serum TNF-alpha concentrations showed a significant decline (65.5-9.75 pg/ml) at the end of the therapy. At the time of discharge from the hospital, there was a positive correlation between neurological recovery and decline in TNF-alpha concentrations in these 22 GBS patients. CONCLUSIONS: The results of this study indicate that elevated levels of TNF-alpha occur in a proportion of patients with GBS and in these patients elevated serum TNF-alpha levels decline with IVIg therapy. PMID- 14742929 TI - Percutaneous vertebroplasty: an experience of 31 procedures. AB - A prospective study of 31 percutaneous vertebroplasty procedures (PVP) in 22 patients treated during January 2000 to December 2001 is presented. PVP was performed using polymethylmethacrylate (PMMA) to treat vertebral collapse due to osteoporosis and vertebral metastasis, to obtain analgesia and spinal stabilization. We analyze the efficacy and complications related to the procedure. PVP is a safe, effective and a daycare surgery. It can be performed under local anesthesia and has minimal and manageable complications. PMID- 14742930 TI - Tropical spastic paraparesis in Kerala. AB - Tropical Spastic Paraparesis (TSP) is an uncommon myeloneuropathy with an insular geographic distribution. In 1985, Human T-lymphotropic virus type I (HTLV-1) was reported to be a possible etiological factor. We did an epidemiological, clinical and virological study of 25 cases of TSP. They were predominantly young adult men, whose duration of illness ranged from 6 months to 15 years. Difficulty in walking, stiffness of legs and paraesthesia of legs were the main symptoms. None had tuberculosis, syphilis, malnutrition or lathyrism. Pyramidal signs of the lower limbs, upper limbs, posterior column involvement and spinothalamic tract dysfunction were the common signs. Laboratory findings and cerebrospinal fluid analysis were normal in most. Myelograms or MRI were normal in all. Only 1 case repeatedly tested positive for HTLV-1 antibodies. None of his relatives had clinical or virological evidence of TSP or HTLV-1 infection. Based on our study, we could not link HTLV-I infection to TSP in Indian cases. Virological testing for HTLV-I infection by polymerase chain reaction may be a better tool to reveal such an association. Our cases were similar to seronegative spinal spastic paraplegia. A related new retrovirus or an altered immune response to HTLV-1 due to environmental or dietary factors are possibilities which require further exploration. PMID- 14742931 TI - Obstructive sleep apnoea: a case-control study. AB - INTRODUCTION: Obstructive sleep apnoea (OSA), is characterized by loud snoring and excessive daytime sleepiness. Though the gold standard for diagnosis is overnight polysomnography (PSG), sleep questionnaires have also been used to diagnose this with good predictive value. MATERIAL AND METHODS: A pre-designed proforma with clinical details, symptom-specific questions for diagnosis of OSA, and Epworth Sleepiness Scale (ESS) was administered to 20 patients presenting to the Sleep Disorder Clinic of our hospital and to 40 age and sex-matched relatives (control group). The students 't-test' and chi-square were used as the statistical tests. RESULTS: There were 20 patients with a mean age of 41+/- 8 years, and 40 controls with a mean age of 41 +/- 6 years (P=>0.05). Seven had family history of snoring in the study group and 3 in the control group (P=0.02). Four had met with road traffic accidents in the study group and none in the control group (P=0.001). The body mass index (BMI) was 29.9 (SD 4.4) in the study group and 24.5 (SD3.5) in the controls (P=0.001). The mean ESS was 13.3 +/- 6 in the patients and 4.2+ 4 in the controls (P=0.001). A larger number of patients with OSA had hypertension: 5/20 vs. 3/40 (P=0.01). CONCLUSION: Patients with OSA had significantly higher BMI and ESS score, and were more likely to have hypertension and road traffic accidents. Increased awareness of this entity is essential. PMID- 14742932 TI - Electrocardiographic findings and prognosis in ischemic stroke. AB - INTRODUCTION: Electrocardiographic (ECG) changes are observed in patients with acute stroke and are related with the prognosis. AIMS: To determine the frequency and significance of ECG changes in patients with acute ischemic stroke. MATERIAL AND METHODS: In a prospective hospital-based study 87 patients with cerebral infarction were observed for ECG changes during their stay in the hospital. All the patients had ischemic stroke for the first time. The ECG changes observed were compared with those of the control group consisting of 87 patients. RESULTS: Of the 87 patients of the study group, 47.1% were females and 52.9% males. The mean age was 65.5 +/- 11.9 (range 31-91 yrs). The control group consisted of 50.6% females and 49.4% males. The mean age was 64.5 +/- 11.9 (range 31-87 yrs). The frequency of the ECG changes observed in patients with cerebral infarct was 62.1% while it was 29.9% in the control group (P<0.0001). ECG changes observed were mostly related to myocardial ischemia. The six-month mortality rate in the patients with ECG changes was 38.9% whereas it was 15.2% in those with normal ECG (P<0.05). CONCLUSIONS: The observations of this study suggest that cardiac evaluation in patients with acute ischemic stroke is of prognostic importance. PMID- 14742933 TI - Radiological and clinical outcome following stereotactic biopsy and radiotherapy for low-grade insular astrocytomas. AB - Thirty patients with low-grade (WHO Grade II) astrocytomas involving the insula, who had undergone stereotactic biopsy followed by radiotherapy, were followed up to evaluate the outcome with regard to control of seizures, memory and language function, Karnofsky Performance scale and regression in tumor volume. Patients were followed up for a mean of 27.8 months, during which time they showed improvement in all the factors that were studied. A statistically significant change was, however, seen only in the reduction in tumor size, probably due to the small sample size and the short duration of follow-up. Stereotactic biopsy followed by radiation therapy provides a good short-term outcome in patients with low-grade insular astrocytomas. PMID- 14742934 TI - Missile injuries of the spine. AB - Between 1995 and 2000, 22 cases with low velocity missile injuries of the spine and spinal cord were treated in three service hospitals. All were adult males, with a mean age of 30.7 years. The wounds were caused by splinters in 18 (82%) and bullets in 4 (18%). Twelve patients received more than one splinter. The cervical and thoracic spines were most frequently involved. In 7 cases, there were injuries to other organs. There was extensive initial deficit (quadriplegia, paraplegia) in 18 (82%) cases, while 4 (18%) had partial deficits. The patients were evaluated by spine radiographs. Myelography was done in 4, CT myelography in 11 and MRI in 4 patients. Two patients had intramedullary hematoma without any skeletal injury, and were treated conservatively. Seventeen patients were treated operatively, and associated injuries of other organs received priority management. Surgery was in the form of debridement, exploration of the spinal cord, hemostasis, decompression and dural repair. Steroids and antibiotics were given routinely. Three patients (2 with cervical and 1 with thoracic spine injury) died preoperatively, and 1 (with dorsolumbar injury) died in the postoperative period due to multi-organ injury. Patients with complete injury remained completely paralyzed, while those with an incomplete injury showed improvement in their neurological grades. The initial neurological grade is the best prognostic indicator, and these injuries are often accompanied by multi organ injuries. There was no instance of postoperative meningitis or CSF leak. These injuries should be explored for debridement and dural repair. PMID- 14742936 TI - Serial pattern learning after mild head injury. AB - Non-verbal serial pattern learning in patients with mild brain injury was examined using a serial pattern-learning task introduced by Nissen and her colleagues. The task involves two types of pattern cycles: Simple and Complex. Each pattern cycle consists of eight acquisition trials followed by a final generation phase, i.e. the ninth trial. The subjects responded to the asterisks appearing in repetitions of a 10-element spatial sequence in each pattern cycle. Eighty subjects were taken, of which 40 subjects had mild head injury and 40 were uninjured. Prior research with this task has shown that individuals show intact performance on the indirect measure of pattern learning, but are impaired on the direct measure. The results of this study showed that mild brain injury does cause a marked disruption in the ability to learn and remember serial pattern information in both simple and complex patterns. PMID- 14742935 TI - Local control of high-grade gliomas with limited volume irradiation versus whole brain irradiation. AB - INTRODUCTION: To evaluate the role of limited field radiation therapy in the management of high-grade gliomas and glioblastoma multiforme (GBM). MATERIAL AND METHODS: From July '96 to January '98, 50 newly diagnosed patients of high-grade gliomas (Grade III and IV) and glioblastoma multiforme who underwent surgery in the form of partial, sub-total or near-total excision as the primary treatment were enrolled in this study. The patients were randomized to receive two different postoperative external radiation protocols, Study Group A: Localized field external radiotherapy 50 Gy/25#/5 wks followed by Boost 10 Gy/5#/1 wk, Control Group B: Whole brain external radiotherapy 40 Gy/20#/4 wks followed by Boost 20 Gy/10#/2 wks by localized field. RESULTS: 20/25 (80%) patients in the study group and 14/25 (56%) patients in the control group showed improvement in their Karnofsky Performance Status (KPS). Thus a significant difference in the performance status was noted in favor of limited field irradiation. No significant difference in the local response was seen between the two groups after radiotherapy. Six months progression-free survival of the study group was 44% as compared to 26% in the control group. Six months overall survival was 66.67% in the study group and 50.72% in the control group (P<0.01). Maximum recurrences were noticed within 2 cm of the original tumor margin in both the groups. CONCLUSIONS: Although local control and survival of the patient in both the groups were same, performance status definitely improved in patients treated with localized field irradiation only. PMID- 14742937 TI - Chordoid meningioma: a report of two cases. AB - Chordoid meningioma is an uncommon histopathological variant of meningioma. We report 2 cases of chordoid meningioma occurring in adult patients. PMID- 14742938 TI - Megalencephalic leukoencephalopathy with subcortical cysts: MRI and proton spectroscopic features. AB - Megalencephalic leukoencephalopathy with subcortical cysts (MLC) is a rare, recently described disease. It is characterized clinically by a large head, mild neurological symptoms and a remarkably slow course of functional deterioration. The MRI is characterized by 'swollen white matter' and large symmetrical cystic changes in the cerebral hemispheres. MLC should be considered in the differential diagnosis of children with megalencephaly and leukoencephalopathy. We report a child with this disease and discuss the MRI and MRS features. PMID- 14742939 TI - Concentric sclerosis: imaging diagnosis and clinical analysis of 3 cases. AB - Balo's Concentric sclerosis (BCS) is a rare demyelinating disease considered to be a variant of multiple sclerosis (MS). The typical magnetic resonance imaging (MRI) changes associated with BCS consist of concentric rings or onions' cross section on T1-weighted (T1W) images. Because MRI reveals pathological changes consistent with autopsy in the focus of BCS, it plays an important role in the before-death diagnosis of BCS. We report three cases of BCS diagnosed antemortem on the basis of the typical concentric rings pattern on MRI and on the basis of clinical findings and cerebrospinal fluid (CSF) examination. BCS often occurs in the prime of life, acutely or subacutely. Then come cerebral multifocal symptoms and signs. We find that BCS is not always an acute and irreversible pathological process as described in the past. PMID- 14742940 TI - Non-convulsive status epilepticus: a rare presentation of juvenile myoclonic epilepsy. AB - We present a case of a boy with juvenile myoclonic epilepsy (JME) who presented with features of non-convulsive status epilepticus (NCSE). This case highlights the fact that NCSE, even though not a common occurrence in JME, should be kept in mind when a patient with previous history of seizures presents with subtle changes in sensorium with no obvious cause. PMID- 14742941 TI - Delayed pharyngo-esophageal perforation: rare complication of anterior cervical spine surgery. AB - An injury to the pharynx and esophagus is a known complication of anterior cervical spine surgery. Two cases of delayed pharyngo-esophageal perforation following anterior cervical spine surgery that resulted in fistula are presented. We postulate that graft displacement and dislodgement of implant with resulting esophageal erosion was responsible for this complication. PMID- 14742942 TI - Traumatic superficial temporal artery aneurysm. AB - Traumatic aneurysm of the superficial temporal artery is an uncommon lesion. Two cases of post-traumatic superficial temporal artery aneurysm are presented. PMID- 14742943 TI - Late postpartum eclampsia without prodroma. AB - Late postpartum eclampsia is an increasingly recognized entity. We describe a patient who developed postpartum eclampsia on the 6th day of delivery without any preceding pre-eclampsia. A high index of suspicion and close follow-up will help in the early detection of this condition. Awareness of this condition will also save the patient from unnecessary investigations. This may be all the more relevant in developing countries where eclampsia contributes to one-third of maternal mortality and the resources for patient investigation and management are limited. PMID- 14742944 TI - Multiple giant congenital melanocytic nevi with central nervous system melanosis. AB - A case of multiple giant congenital melanocytic naevi in whom central nervous system melanosis was detected at 6 weeks of age is described. The infant was asymptomatic, but presence of risk factors such as multiple naevi, giant naevi and naevi on scalp and posterior axial location prompted a magnetic resonance imaging study of the brain. To our knowledge, neurocutaneous melanosis at such a young age has not been reported in Indian literature. PMID- 14742945 TI - Report of five children with Guillain-Barre syndrome following a nationwide oral polio vaccine campaign in Turkey. AB - Five children with Guillain-Barre syndrome (GBS), following a national oral polio vaccination campaign to eradicate disease, are reported. Clinical examination, cerebrospinal fluid and electromyographic findings conformed to the classical description of GBS. Four of them received therapeutic dose of intravenous immunoglobulin G. Two children succumbed to the disease. It was observed that the number of cases of GBS in children increased during the period of the oral polio vaccination campaign in Turkey, suggesting a causal relationship. PMID- 14742946 TI - Medulloepithelioma: a case report. AB - Histological, immunohistochemical, and CT morphological features of medulloepithelioma, a rare embryonal tumor of primitive neuroepithelium, are described. PMID- 14742947 TI - Pituitary tuberculoma: magnetic resonance imaging. AB - A case report of a pituitary tuberculoma, with magnetic resonance imaging (MRI) features is described. The patient was treated with anti-tuberculous chemotherapy, and a follow-up MRI after 6 months showed good response. The relevant literature is briefly reviewed. PMID- 14742948 TI - Neurocysticercosis presenting as Weber's syndrome. AB - This case report describes a rare, non-epileptic manifestation of neurocysticercosis where a 22-year-old male presented with acute onset right 3rd nerve palsy with left hemiplegia (Weber syndrome). Computerized tomography and magnetic resonance imaging revealed cysticercus granuloma. The patient improved and became asymptomatic with steroid treatment. Recognizing this clinical entity would avoid unnecessary antituberculous treatment and surgical intervention. PMID- 14742949 TI - Tapping of a type II Sylvian fissure arachnoid cyst: technical note. AB - A 28-year-old man with a large Sylvian fissure cyst was treated by making a small pre-coronal burr hole, and subsequently, under a direct view its wall was fenestrated with a Cushing's needle and the cyst fluid was tapped. The patient had complete neurological recovery. The follow-up was of 90 months. PMID- 14742950 TI - Steroid-induced myopathy following a single oral dose of prednisolone. AB - This report describes a case of acute steroid-induced myopathy following a single dose of oral prednisolone. A 55-year-old man presented with an acute exacerbation of chronic obstructive pulmonary disease, which was treated with prednisolone 40 mg daily in addition to bronchodilators. He developed features of myopathy the next day. Serum CPK was moderately elevated and electromyogram was suggestive of primary muscle disease. He was managed conservatively and improved 10 days after stopping prednisolone. Mechanisms of steroid-induced myopathy and relevant literature have been reviewed. PMID- 14742951 TI - Cerebrotendinous xanthomatosis with oromandibular dyskinesia. AB - We present an unusual case of cerebrotendinous xanthomatosis in a female elderly patient with recurrent TM joint dislocation and oromandibular dyskinesia. PMID- 14742952 TI - Guillain-Barre syndrome presenting in the anti-HIV seroconversion period. PMID- 14742953 TI - Mechanical ventilation in Guillain-Barre syndrome. PMID- 14742955 TI - Airway safety for patients receiving intraventricular sodium nitroprusside therapy. PMID- 14742956 TI - Factors affecting functional recovery in ischemic stroke. PMID- 14742957 TI - Stereotactic brainstem biopsy is indicated for the diagnosis of a vast array of brainstem pathology. AB - BACKGROUND/AIMS: A meta-analysis of thirteen studies performing stereotactic biopsy of brainstem lesions is combined with our recent series of 12 patients with brainstem lesions comparing MRI findings to biopsy histopathology to determine whether these lesions could have been diagnosed radiographically. METHODS: 12 consecutive stereotactic biopsies and literature analysis were performed to analyze clinical, radiographic and histopathologic data. RESULTS: Stereotactic biopsy of lesions in the brainstem was in 96% diagnostic. There was one mortality (3% of cases) and 4% morbidity associated with the procedure. Pathology showed that half of the adult brainstem intrinsic lesions were gliomas, 10% were metastases, and the remainder were hematomas, vascular malformations, lymphomas, demyelination, cysts, radiation necrosis, abscesses, vasculitis, infarcts, leukemia, cryptococcus, or granulomas. CONCLUSIONS: Empiric treatment of adult brainstem lesions is not prudent because there is a wide spectrum of diverse pathology in this location. Stereotactic biopsy is a safe and effective method for determining histopathology. PMID- 14742958 TI - Permanent iodine-125 interstitial radiation therapy in the treatment of non glioblastoma multiforme high-grade gliomas. AB - BACKGROUND: This study evaluates prognostic factors influencing survival outcomes for 60 patients with permanent iodine-125 implants in the primary treatment of non-glioblastoma multiforme (GBM) high-grade gliomas. METHODS: Stereotactic treatment planning aimed to encompass the contrast-enhancing rim of the tumor visualized by CT, with an initial dose rate of 0.05 Gy/h with 125I, delivering 100 Gy at 1 year and 103.68 Gy at infinity. Survival was evaluated using the Kaplan-Meier method for univariate analysis and the Cox regressional method for multivariate analysis. In addition to the implant, 34 patients received external radiation therapy (5,000-6,000 cGy) before the implant; 13 patients were implanted without additional external beam radiation, and 13 patients underwent external radiation therapy before implant placement. RESULTS: With a mean follow up of 77.6 months (range 3.5-164 months), 1-, 3-, 5- and 10-year survival were 86.7% (+/-0.05%), 60% (+/-0.07%), 50% (+/-0.07%) and 45.7% (+/-0.7%), respectively. The median survival time was 57 months. Second surgery was performed following the implant in 19 patients. Findings were tumor recurrence in 11 patients (22.5%), radiation necrosis in 7 patients (14.3%) and brain abscess in 1 patient (2%). Age, sex, tumor location, side of brain, tumor volume, Karnofsky score and neurological status were correlated with survival outcome. Favorable prognostic factors were age younger than 45 years, superficial tumor location and preoperative Karnofsky score greater than 70. RPA classification was used to define this group of patients. In RPA classes I and II (n = 43), 1-year survival was 93%, while 3-, 5- and 10-year survival was 67.4, 60.5 and 55.5%, respectively, and median survival time was 91 months. In RPA class III (n = 7), 1 year survival was 71.4%, while 3- and 5-year survival was 42.9 and 28.6%, respectively, and median survival time was 47 months. In RPA class IV (n = 10), 1 year survival was 60%, while 3-, 5- and 10-year survival was 50, 22.2 and 11.1%, respectively, and median survival time was 37 months. CONCLUSION: Brachytherapy with permanent implant of 125I appears promising in the treatment of primary non GBM malignant gliomas. It improved survival time and reduced the incidence of complications and provided good quality of life. In order to further confirm these results, multicenter randomized prospective studies are needed. RPA analysis is a valid tool to define prognostically distinct survival groups. In this study, 2-year survival and median survival time were improved in all prognostic classes. This would suggest that selection bias alone does not account for the survival benefit seen with 125I implants. Further randomized studies with effective stratification are needed. PMID- 14742959 TI - Posterior fossa metastases: aggressive treatment improves survival. AB - BACKGROUND: Brain metastases are a leading cause of mortality and morbidity in patients with malignancies. Infratentorial location has been considered a negative prognostic factor. METHODS: This retrospective study evaluated patients with cerebellar metastasis. Statistical analysis assessed age, extracranial disease, performance status and treatment. Patients were categorized by Radiation Therapy Oncology Group recursive partitioning analysis (RPA). Treatment included surgery, stereotactic radiosurgery (SRS) and whole brain radiotherapy (WBRT) alone or in combination. RESULTS: Of 93 patients, the median survival was 12.9 months for RPA class I, 11 months for class II and 8 months for class III. On multivariate analysis, RPA class was an important predictor for overall survival. However, SRS with WBRT or surgery with WBRT or a combination of SRS, surgery and WBRT, was more favorable than surgery or SRS alone within RPA class II patients. CONCLUSIONS: Survival of patients with cerebellar brain metastasis is comparable to that of patients with supratentorial brain metastasis using RPA classification. Aggressive multimodality therapy has a favorable impact on survival. PMID- 14742960 TI - Interstitial irradiation for CNS lesions. AB - In this study, we investigated the efficacy of brachytherapy in the treatment of 138 patients with intracranial neoplasms of the CNS. Of the total number of patients, 50 presented with glioblastoma multiforme, 45 presented with low-grade glioma, 19 presented with anaplastic astrocytoma, 23 presented with metastases and 1 presented with meningioma. During the execution of this study, seeds of 125I (10-20 mCi) were inserted into the lesions to aim the irradiation at a low dose of 60 Gy in the margin of benign lesions or 1 cm beyond the radiological border of malignant lesions, which were visualized on CT scan. The results of this procedure were evaluated in terms of the survival rates, which were assessed by Kaplan-Meier curves. Significant relationships were not observed between the volume and location of the lesions, the whole-brain radiotherapy and chemotherapy, and the survival time of the patients. A low Karnofsky Index score and older age were associated with a short survival time. In light of the above, it was concluded that interstitial irradiation is a safe and effective method of treatment for brain tumors. PMID- 14742961 TI - From letterbox to keyhole approach for resecting intracranial lesions. AB - BACKGROUND: Computer-assisted neurosurgery systems (CANS) have the ability to translate preoperative image data sets directly to the operating field and were thought to be very useful in neurosurgery. However, key questions regarding their use remain only partially addressed. METHODS: To answer the doubts of the skeptics, we set up a registry of all CANS and any non-CANS procedures in our institution. The results of 354 procedures are presented in this paper. RESULTS: CANS was used in 254 procedures, with a mean accuracy of 1.9 mm and a failure rate of 2.8%. Over time, the accuracy improved to 1.6 mm and failures were abolished. The CANS was most useful in anterior skull approaches and took less time in the operating room, and the patients were discharged earlier than those who underwent non-CANS procedures. CONCLUSION: We feel that the introduction of CANS has changed our practice for the better, with significant benefits to patients, surgeons and the hospital at large, and we recommend its usage as a standard of care. PMID- 14742962 TI - Bulbar trigeminal stereotactic nucleotractotomy for treatment of facial pain. AB - Many pharmacological and surgical treatments are available for the treatment of chronic facial pain. However, many of them are expensive and often very ineffective. Past publications suggested that bulbar trigeminal stereotactic nucleotractotomy is a very useful procedure for the treatment of neuropathic or oncologic facial pain. The authors describe the results of treatment with stereotactic nucleotractotomy in 58 patients with chronic facial pain. The intensity of the pain was evaluated according to the visual analogue scale, and daily life activities were also evaluated. The conclusion was that this procedure is a safe and effective method for treatment of postherpetic neuralgia, Wallenberg's syndrome and oncologic facial pain but not of trigeminal neuralgia. PMID- 14742963 TI - Repeat gamma knife radiosurgery for trigeminal neuralgia. AB - BACKGROUND: Although gamma knife radiosurgery (GKRS) has been shown to be safe and effective for the treatment of trigeminal neuralgia (TN), there are few studies that report the results of a second GKRS. METHOD: Between May 22, 1998 and April 1, 2003, we treated 335 primary TN patients with GKRS. All received a maximum dose of 75 Gy to the cisternal trigeminal nerve. 45 patients with recurrent or persistent TN were treated with a maximum dose of 40 Gy at a second GKRS and were available for at least 6 months of follow-up. RESULTS: Final pain relief (mean of 15 months after second GKRS) was 50% or greater in 28 of the 45 patients (62.2%). Patients who had no neurosurgical procedure prior to their first GKRS were more likely to have pain relief of 50% or greater following the second GKRS (p = 0.042). Significant new dysesthesias (score greater than 5 on a scale of 0-10) developed in 2 patients (4.4%). CONCLUSION: Repeat GKRS has a good chance of relieving TN pain without complications and is more likely to relieve pain in those who did not have any procedure prior to their first GKRS. PMID- 14742964 TI - Long-term outcome of spinal cord stimulation and hardware complications. AB - Spinal cord stimulation (SCS) is a treatment modality for medically intractable chronic pain. This study reports an 11-year experience with SCS assessing long term pain relief and specifically evaluating complications and revisions. It took the form of a retrospective review of medical/surgical records with a postal questionnaire. The subjects were 102 patients with medically intractable chronic pain who underwent SCS implantation between 1989 and 2000. There were 64 revision operations carried out on 35 patients. These comprised electrode replacement/repositioning (29), generator replacement (23), cable failure (3) and implant removal (5). Five (4.9%) implants became infected and 2 required removal. Clinician-reported pain relief was substantial in 69 (68%) patients. This study adds to the weight of evidence that patients undergoing SCS derive significant benefits in terms of pain relief. However, revision rates remain high due to technical and biological factors. PMID- 14742965 TI - Microvascular decompression for trigeminal neuralgia in patients with multiple sclerosis. AB - AIMS: To assess whether microvascular decompression (MVD) is a safe and efficacious treatment for patients with trigeminal neuralgia (TGN) and multiple sclerosis (MS). METHODS: Case records were reviewed of all patients with TGN and MS who underwent posterior fossa exploration with a view to MVD between 1993 and 2001. In all patients, magnetic resonance tomoangiography (MRTA) demonstrated vascular compression. RESULTS: Nine patients were included in the study. Seven patients underwent MVD alone; in 2 patients a partial sensory rhizotomy of the trigeminal nerve was added to the decompression. All patients had excellent initial pain relief. Recurrence of neuralgia was noted in 5 patients after MVD and in 1 of the 2 patients after partial sensory rhizotomy. Long-term pain relief was obtained in 1 patient who underwent a redo MVD after postoperative MRTA scans demonstrated recurrent vascular compression of the root entry zone (REZ). Thus only 4 out of 9 patients maintained pain relief. In addition, 3 patients experienced transient worsening of their MS. CONCLUSIONS: Although MVD provides good initial pain relief, the recurrence rate is much higher than that obtained in 'idiopathic' TGN. Although all procedures for the treatment of TGN are worse than those for idiopathic TGN, it is concluded that because of the high recurrence rate together with the morbidity associated with the procedure MVD should not be offered to patients with TGN and MS. PMID- 14742966 TI - Chronic pain after spinal cord injury: clinical characteristics. AB - The clinical characteristics of chronic pain in spinal cord injury patients are controversial. The authors prospectively evaluated 81 patients with chronic pain due to spinal cord lesions. The mean pain intensity according to the visual analogue scale was 9.4. The most common description of pain was a sensation of burning. The initial pain was more severe in patients presenting with myelopathy due to gunshot injuries (p < 0.001). The pain intensity was not associated with the magnitude of the spinal lesion, location of the lesion, occurrence of myofascial pain syndrome or onset of pain. Pain after spinal cord injury was severe, males were more frequently affected and it was more intense when it was the result of gunshot injury. In about 38% of the patients, pharmacological and rehabilitative procedures were effective. Dorsal root entry zone lesion was effective for the treatment of transitional pain in patients with complete section of the spinal cord, spinal cord stimulation was effective for patients with partial lesions of the spinal cord and intrathecal opioid infusion was effective for both conditions. PMID- 14742967 TI - Spinal cord stimulation for the treatment of refractory unilateral limb pain syndromes. AB - BACKGROUND: Spinal cord stimulation (SCS) is an established therapy for chronic pain. Its success depends on vigorous patient selection and good follow-up. METHODS: We reviewed 75 patients who had undergone SCS to establish their outcome. 67 of these patients had refractory unilateral limb pain syndrome (RULPS). Their case notes and operative log books were critically reviewed, and when appropriate, telephone interviews were performed (58 patients). RESULTS: 87% of patients responded initially to SCS; at 6 months, the effect waned to 79%, and by 2 years, it improved to 84%. One third of the patients had no revisions, 40% had IPG replacements and the rest had revisions because of lead-related complications (5.3%), epidural complications (mainly fibrosis; 19%) or infections (2.7%). 56% of patients reduced their analgesia, 1.5% stopped taking any painkillers and 46.8% of those who were employed returned to work. CONCLUSION: We feel that SCS is an effective treatment in RULPS and its results depend upon vigorous patient selection and an adequate follow-up and maintenance program. PMID- 14742968 TI - Role of pituitary radiosurgery for the management of intractable pain and potential future applications. AB - RATIONALE: Two or three decades ago, cancer pain was treated by surgical/chemical hypophysectomy. In one report, the control of central pain (thalamic pain syndrome) was also approached with chemical hypophysectomy. Although in most of the patients these treatments resulted in a decrease in severe pain, concomitantly severe adverse effects (panhypopituitarism, diabetes insipidus and visual dysfunction) occurred in most patients. This historical evidence prompted us to perform Gamma Knife surgery (GKS) for this kind of intractable severe pain using a high irradiation dose to the pituitary stalk/gland. In the majority of patients, marked pain relief was achieved, surprisingly without any of the complications mentioned above. MATERIALS AND METHODS: A prospective multicenter study was conducted to evaluate the efficacy and safety in patients treated in Prague, Hong Kong and Tokyo. Indications of this treatment were: (1) failure of other effective treatment approaches prior to GKS, (2) good general patient condition (Karnofsky performance status >40%), (3) response to morphine for pain control (cancer pain), and (4) no previous radiotherapy of brain metastases (GKS/conventional radiotherapy). Eight patients with severe cancer pain due to bone metastasis and 12 patients with post-stroke thalamic pain syndrome were treated with GKS. The target was the border between the pituitary stalk and gland. Maximum dose was 160 Gy for cancer pain and 140 Gy for central pain. Follow-up included 6 patients (>1 month) with cancer pain and 8 patients (> 6 months) with thalamic pain syndrome. RESULTS: All patients (6/6) with cancer pain experienced significant pain reduction, and 87.5% (7/8) of the patients with thalamic pain had initially significant pain reduction. In some patients, pain reduction was delayed for several hours. Pain relief was noted within 7 days (median 2 days). No recurrence was observed in the patients with cancer pain. However, in 71.4% (5/7) of the patients with thalamic pain syndrome, disease recurred during the 6-month follow-up. Up to now, other complications have not been observed. CONCLUSION: Our clinical study protocol is only preliminary. Further clinical results on the management of thalamic pain are required to develop this treatment protocol. However, efficacy and safety have been shown in all our cases. In our opinion, this treatment has a potential to control severe pain, and GKS will play an important role in the management of intractable pain. PMID- 14742969 TI - Evaluation of CyberKnife frameless real-time image-guided stereotactic radiosurgery for spinal lesions. AB - BACKGROUND: This study evaluated the CyberKnife frameless image-guided radiosurgery system for the treatment of spinal lesions. METHODS: This system utilizes the coupling of an orthogonal pair of X-ray cameras to a dynamically manipulated robot-mounted linear accelerator that guides the therapy beam to the intended target without the use of frame-based fixation. Cervical spine lesions are located and tracked relative to skull bony landmarks; lower spinal lesions are tracked relative to fiducial markers. 125 spinal lesions in 115 consecutive patients were treated with a single-fraction radiosurgery technique. RESULTS: Tumor volume ranged from 0.3 to 232 ml (mean 27.8 ml). Tumor radiation dose was maintained at 12-20 Gy to the 80% isodose line (mean 14 Gy); the spinal cord or canal volume receiving greater than 8 Gy ranged from 0.0 to 1.7 ml (mean 0.2 ml). No acute radiation toxicity or new neurological deficits occurred during the follow-up period (3-24 months). CONCLUSIONS: The CyberKnife system was found to be feasible, safe and effective. The major potential benefits of radiosurgical ablation of spinal lesions are short treatment time in an outpatient setting with rapid recovery and good response. PMID- 14742970 TI - Bilateral anterior capsulotomy for refractory obsessive-compulsive disorders. AB - A prospective study on the effects of bilateral anterior capsulotomy in patients with refractory obsessive-compulsive disorder (OCD) is presented. A total of 18 procedures were performed in 15 patients. The mean duration of the OCDs was 18.1 +/- 5.6 years (range 11-26 years). The mean total Yale-Brown Scale (Y-BOCS) score was 29.67 and mean Global Assessment of Functioning was 43.61. The results throughout the follow-up period remained the same as at 1 month postoperatively. The observed mean recovery on the Y-BOCS was 33.2% (p = 0.017). 52.9% of the patients showed a 33% recovery, 29.4% of the cases showed a 50% recovery and 17% showed a 66% recovery. Global Assessment of Functioning recovered by 19% (p = 0.111). No cognitive deficit was disclosed by neuropsychological screening tests. Complications were observed in 3 cases, 1 with transitory hallucinations, 1 with a single epileptic seizure and 1 case who developed a progressive behavior disorder that became permanent. We conclude that bilateral anterior capsulotomy is a safe and effective procedure. PMID- 14742971 TI - Clinical study for alleviating opiate drug psychological dependence by a method of ablating the nucleus accumbens with stereotactic surgery. AB - The aim of this study was to explore a new way of treating drug addiction by ablating the nucleus accumbens (NAC), which has a close relationship with drug induced psychological dependence, using stereotactic surgery, blocking the mesocorticolimbic dopamine circuit, alleviating craving for drugs and lowering the relapse rate after detoxification. On the basis of animal experiments, stereotactic surgery was performed in 28 patients by making a lesion in the NAC bilaterally to treat opiate drug dependence. Indications, the criterion of therapeutic effect, treatment process and the therapeutic and safety evaluation index of the surgery were formulated particularly. The mean follow-up period was 15 months. Relapse has not occurred in 11 cases up till now. Drug-free time in these patients has been more than half a year in 4 cases (more than a year in 3 cases), and less than half a year in 7 cases. Relapse occurred in 15 cases after surgery. Drug-free time in these patients was more than half a year in 3 cases, between 1 month and half a year in 10 cases and less than 1 month in 2 cases. The therapeutic effect was excellent in 7 cases (26.9%), good in 10 cases (38.5%) and poor in 2 cases (7.7%). Another 7 cases were still under investigation at the time of writing. Relapse rates after surgery were 7.7, 38.5 and 57.5% within 1 month, between 1 month and half a year and after more than half a year, respectively. There were no common complications of surgery such as intracranial hematoma or infection in these patients after operation. Character type was changed slightly in 2 cases, and 4 cases suffered temporary memory loss, which did not affect their daily lives and learning function. They all recovered within 1 month. There were different degrees of effectiveness of treating drug addicts' psychological dependence by making lesions in the NAC bilaterally with stereotactic surgery. No particular complications occurred. The operation is safe and feasible. The mean follow-up time in this study was 15 months. The effectiveness was satisfactory. The relapse rate of drug addicts after detoxification was clearly reduced. PMID- 14742972 TI - Cyberknife radiosurgery for trigeminal neuralgia. AB - BACKGROUND: We present preliminary results using Cyberknife radiosurgery as a noninvasive treatment for trigeminal neuralgia (TN). METHODS: Ten patients with medically refractory TN who were deemed unsuitable for conventional surgery underwent Cyberknife radiosurgery using CT cisternography for localization. RESULTS: Pain relief was achieved in 7 patients, in 5 of them within 24-72 h after irradiation. CONCLUSION: Cyberknife radiosurgery can achieve early-onset pain relief in a subset of TN patients. Improvements using this technique include the absence of a stereotactic ring, potentially improved targeting accuracy produced by CT cisternography and improved dose homogeneity. PMID- 14742973 TI - Use of a multichannel auditory brainstem implant for neurofibromatosis type 2. AB - Neurofibromatosis type 2 (NF 2) typically results in deafness due to disruption of the cochlear nerves, making peripheral devices such as cochlear implants ineffective. Auditory brainstem implants (ABIs), for direct electrical stimulation of the cochlear nucleus, have been used to provide auditory stimulation in this group of patients. Currently, 141 patients have been implanted in our institution, most recently using an advanced multichannel device. We report results of a recent series of 86 patients who received ABIs. Of this group, 60 had successful implantation, recovered from surgery, responded successfully to stimulation and underwent a full course of device programming and audiologic testing. This group had significant improvement in scores on several audiologic tests compared to baseline. When used to augment lip reading, improvement was also seen. The degree of improvement varied considerably among patients. ABI is a useful device for deaf patients with NF 2. As measured by audiologic testing, many patients receive substantial benefit with regard to sound and speech comprehension. PMID- 14742974 TI - Predictive factors of radiation necrosis after radiosurgery for cerebral metastases. AB - The aim of this study was to determine if the risk of radiation necrosis after radiosurgery is related to the presence of normal tissue included in the prescription volume. Between 1994 and 2001, 377 patients representing 760 lesions were treated by radiosurgery in our center with a 10-MV LINAC. The median age of the patients was 57 years (range 30-86 years), median tumor volume was 4.9 cm(3), median peripheral dose (70%) was 15.6 Gy and median dose at the isocenter was 21.6 Gy. Karnofsky index, disease control and number of lesions were the only parameters significantly influencing survival (median 8.6 months), while disease free survival was correlated with the number of isocenters. Seven percent of the patients presented severe complications, including nine episodes of radiation necrosis. The only parameter influencing the risk of radiation necrosis was the conformity index (p = 0.001). These findings emphasize the importance of reducing falsely irradiated normal tissue during radiosurgery to prevent radiation necrosis. PMID- 14742975 TI - A meta-analysis from the Cochrane Library reviewing interleukin 2 receptor antagonists in renal transplantation. PMID- 14742976 TI - Interleukin 2 receptor antagonists for renal transplant recipients: a meta analysis of randomized trials. AB - BACKGROUND: Interleukin 2 receptor antagonists (IL-2Ra) are increasingly used to treat renal transplant recipients. This study aims to systematically identify and summarize the effects of using IL-2Ra as induction immunosuppression, as an addition to standard therapy, or as an alternative to other antibody therapy. METHODS: Databases, reference lists, and abstracts of conference proceedings were searched extensively to identify relevant randomized controlled trials in all languages. Data were synthesized using the random effects model. Results are expressed as relative risk (RR), with 95% confidence intervals (CI). RESULTS: A total of 117 reports from 38 trials involving 4,893 participants were included. When IL-2Ra were compared with placebo (17 trials; 2,786 patients), graft loss was not significantly different at 1 year (14 trials: RR 0.84; CI 0.64-1.10) or 3 years (4 trials: RR 1.08; CI 0.71-1.64). Acute rejection was significantly reduced at 6 months (12 trials: RR 0.66; CI 0.59-0.74) and at 1 year (10 trials: RR 0.67; CI 0.60-0.75). At 1 year, cytomegalovirus infection (7 trials: RR 0.82; CI 0.65-1.03) and malignancy (9 trials: RR 0.67; CI 0.33-1.36) were not significantly different. When IL-2Ra were compared with other antibody therapy, no significant differences in treatment effects were demonstrated, but IL-2Ra had significantly fewer side effects. CONCLUSIONS: Given a 40% risk of rejection, seven patients would need treatment with IL-2Ra in addition to standard therapy, to prevent one patient from undergoing rejection, with no definite improvement in graft or patient survival. There is no apparent difference between basiliximab and daclizumab. PMID- 14742977 TI - Administration of nitric oxide with caspase inhibitors minimizes bacterial translocation in experimental intestinal transplantation. AB - BACKGROUND: Bacterial translocation (BT) has been suggested to be responsible for the high incidence of infections occurring after small-bowel transplantation (Trp). Nitric oxide (NO) and apoptosis could affect cell demise. The aim of this study was to asses whether supplementation of University of Wisconsin (UW) solution with NO donors and apoptosis inhibitors can abolish BT in Trp. METHODS: The following experimental groups were studied: sham, Trp, intestinal transplantation, Trp+spermine NONOate (NONOs), and Trp+NONOs+caspase inhibitor Z Val-Ala-Asp(Ome)-fluoromethylketone(Z-VAD-fmk). Histologic analysis, caspase-3 activity, DNA fragmentation, and BT from graft to mesenteric lymph nodes, liver, and spleen were measured in tissue samples after transplantation. RESULTS: During intestinal transplantation, apoptosis and necrosis were increased, showing graft injury and high levels of BT. The rats treated with NONOs showed a histologic protection of transplanted graft and a decrease in BT despite caspase-3 and DNA fragmentation-inducing effects. Administration of caspase inhibitor Z-VAD to NONOs-treated rats reversed the NO apoptosis-inducing effects and showed the lowest levels of BT in all tissues. CONCLUSIONS: Exogenous administration of NO associated with the inhibition of apoptosis maintains the graft in optimal conditions in terms of BT and improves the histology of the graft after intestinal transplantation in rats. PMID- 14742978 TI - Acceleration of apoptosis in CD4+CD8+ thymocytes by rapamycin accompanied by increased CD4+CD25+ T cells in the periphery. AB - BACKGROUND: Rapamycin (Rapa) is an immunosuppressant that is used in patients and animal models to control allograft rejection. Its mechanisms of action are not fully understood. In this article, the authors have investigated the effects of therapeutic doses of Rapa on both thymic and peripheral T-cell populations in the adult rat. METHODS: The therapeutic dosage of Rapa was optimized using cardiac transplantation between LEW and DA rats. Thymic morphology was assessed by hematoxylin-eosin staining. Flow cytometric analysis was performed to analyze T cell phenotype and apoptosis. T-cell receptor (TCR)-mediated T-cell responsiveness was evaluated by 3[H]-thymidine deoxyribose incorporation. RESULTS: Rapa induced atrophy in the thymus but not in peripheral lymphoid organs. Moreover, fibrosis occurred in thymus that was long-lasting after Rapa withdrawal. In animals treated with Rapa, there was a significant reduction in CD4+CD8+ thymocytes caused by accelerated apoptosis, whereas CD4-CD8-, CD4+CD8-, and CD8+CD4- populations remained unaffected. In contrast, the cellularity of the periphery lymphoid organs was not altered. Within the CD4+ thymocyte population, CD4+CD25+ thymocytes were resistant to Rapa-accelerated apoptosis, and in the periphery, the ratio of CD4+CD25+ to CD4+CD25- T cells was increased. Notably, the peripheral CD4+CD25+ T cells were hyporesponsive to TCR-mediated activation. CONCLUSIONS: The resistance of the peripheral CD4+CD25+ T cells to Rapa treatment might contribute to its immunosuppressive action. The long-term effects of Rapa on thymus atrophy and thymocyte development requires consideration with respect to its clinical application. PMID- 14742979 TI - Treatment of recurrent hepatitis C infection after liver transplantation with combination of pegylated interferon alpha2b and ribavirin: an open-label series. AB - BACKGROUND: Hepatitis C virus (HCV) recurrence after orthotopic liver transplantation (OLT) is universal. We aimed to evaluate the efficacy and safety of pegylated interferon (PEG-IFN) and ribavirin (RIB) in the treatment of post OLT HCV recurrence. METHODS: Thirty-seven patients with recurrent HCV after OLT were screened and began treatment. Nineteen patients have completed therapy. PEG IFN was started at a dose of 0.5 microg/kg per week and titrated toward a maximum dose of 1.5 microg/kg per week. RIB was started at a dose of 400 mg per day and titrated toward a maximum of 1000 mg per day, as tolerated. Therapy continued for 1 year after HCV replication was undetectable by reverse transcriptase-polymerase chain reaction and was discontinued if there was no virologic clearance at 48 weeks. RESULTS: Twelve patients (63%) completed the combination regimen. Therapy was discontinued in seven (37%) patients. Seven patients (37%) had undetectable viral load at the end of treatment. Of those, five patients (26%) had sustained viral response 6 months after discontinuation of therapy. Five patients (26%) had no virologic response. Necro-inflammatory score declined from 5.22 to 2.89 (P=0.05) in nonresponders versus 6.8 to 2.6 (P<0.01) in responders. Fibrosis stage did not change in either group. Genotype 1-infected patients had a lower likelihood of attaining end of treatment or sustained viral response (P<0.05 for both). CONCLUSIONS: Post-OLT HCV recurrence can be safely treated with PEG-IFN and RIB. Bone marrow toxicity, depression, and rejection are limiting factors that require aggressive management. There was short-term histologic benefit to the use of this regimen, even in those patients without viral clearance. PMID- 14742980 TI - Increased immune transcript levels are correlated with acute graft-versus-host disease and cytomegalovirus response after allogeneic stem cell transplantation. AB - BACKGROUND: Donor T cells are primarily responsible for graft-versus-host disease (GVHD). Three effector pathways have been described for T-cell cytotoxicity: granzyme B/perforin, Fas/Fas ligand (FasL), and secreted molecules such as tumor necrosis factor (TNF)-alpha. Therefore, this study evaluates the gene expression pattern in the peripheral blood of patients after allogeneic stem cell transplantation and correlates the results to acute GVHD. METHODS: Real-time quantitative reverse transcriptase-polymerase chain reaction was used to quantify the gene expression of granzyme B, perforin, FasL, and TNF-alpha in peripheral blood from 53 patients. RESULTS: Samples were available from 27 of the 38 patients with acute GVHD diagnoses. Increased gene expression (>50%) during acute GVHD was detected in 23 of 27, 26 of 27, and 24 of 27 patients for granzyme B, perforin, and FasL, respectively. TNF-alpha showed a diffuse correlation. The median increases were as follows: granzyme B, 7.2x (1.6-183.2); perforin, 5.8x (1.6-254.9); and FasL, 8.5x (1.5-895.6). We also showed that all of the 10 patients with increasing levels of granzyme B, perforin, and FasL during steroid treatment demonstrated persistent or deteriorating GVHD. Patients with increasing transcription levels during cytomegalovirus (CMV) reactivation responded significantly better to therapy than those with declining levels. A total of 13 of 17 patients with increasing levels versus 0 of 11 patients with decreasing levels responded well to CMV treatment (P<0.01). CONCLUSION: Although not specific for acute GVHD, quantitative assessment of immune transcripts may be of value in diagnosing and monitoring acute GVHD. It may also serve as a guide for the clinician in detecting patients who respond poorly to CMV therapy. PMID- 14742981 TI - Biochemical prognostic markers of outcome in non-paracetamol-induced fulminant hepatic failure. AB - BACKGROUND: Fulminant hepatic failure (FHF) is associated with major metabolic disturbances, the onset and severity of which can predict clinical outcome. This study uses admission blood samples to identify early biochemical markers of clinical outcome in patients with non-paracetamol-induced FHF. PATIENTS AND METHODS: Fifty-nine patients admitted to the Scottish Liver Transplant Unit with non-paracetamol-induced FHF were studied. Plasma samples were collected at a median of 5.4 hr after admission to our unit and analyzed using conventional laboratory tests and nuclear magnetic resonance spectroscopy. RESULTS: A total of 19 patients underwent transplantation, 15 patients died without undergoing transplantation, and 25 patients survived with medical management alone. There were significantly lower levels of lactate, alanine, valine, and bilirubin and significantly higher levels of pyruvate and albumin in patients who survived spontaneously compared with the other two groups. By use of multiple logistic regression analysis, an equation was devised that best predicted clinical outcome: 0.5x(albumin [g/L])-2x(lactate [mmol/L])-36x(valine [mmol/L]) 38x(pyruvate [mmol/L]). Values of less than 2 were associated with poor clinical outcome and had a positive predictive value of 91%, a negative predictive value of 86%, a sensitivity of 94%, and a specificity of 86% for death or transplantation. This algorithm can be applied on admission, thus expediting decision-making. CONCLUSION: We identified biochemical markers that may be useful in predicting outcome in patients with non-paracetamol-induced FHF and should be evaluated further in a different patient population. PMID- 14742982 TI - The effect of oral iron admiinistration on mycophenolate mofetil absorption in renal transplant recipients: a randomized, controlled trial. AB - BACKGROUND: Oral iron supplements are frequently prescribed to renal transplant recipients in the early posttransplant period. A recent trial in seven healthy volunteers demonstrated a significant 91% reduction in mycophenolate mofetil (MMF) absorption when coadministered with oral iron. However, the effect of iron on MMF absorption in renal transplant patients has not been studied. METHODS: An open-label, randomized, controlled trial was undertaken in which new renal transplant recipients were randomly allocated to receive iron supplements with a morning dose of MMF, iron supplements given 4 hr after MMF at midday, or no iron supplements. Blood samples were taken for estimation of mycophenolic acid (MPA) area under the curve (AUC) at day 5 posttransplant. The primary endpoint was the day 5 MPA AUC, with secondary endpoints including acute rejection and MMF toxicity in the first 4 weeks posttransplant. Prospective power calculations indicated that a minimum of 13 patients in each group would be required to have a 90% probability of detecting a clinically significant reduction (10 mg/hr/L) in MPA AUC for iron-treated patients. RESULTS: Forty patients completed the study. There were no differences in baseline demographic data between the groups. The mean+/-standard deviation MPA AUC measurements for the groups receiving no iron (n=13), iron and MMF together (n=14), and iron and MMF spaced apart (n=13) were 34.5+/-8.7, 33.7+/-11.4, and 32.1+/-8.1 microg/hr/mL, respectively (P=0.82). Rates of acute rejection, cytopenia, infection, and gastrointestinal intolerance were comparable between the groups. CONCLUSIONS: There is no significant effect of oral iron supplements on MMF absorption as determined by measured blood concentrations. The practice of routinely giving oral iron in such patients seems safe from an immunosuppression drug-interaction standpoint. PMID- 14742983 TI - Hepatitis C infection-related liver disease: patterns of recurrence and outcome in cadaveric and living-donor liver transplantation in adults. AB - BACKGROUND: Preliminary data demonstrate that the recurrence of hepatitis C is more severe in patients undergoing adult-to-adult living liver (AAL) transplantation (Tx) in comparison with cadaveric liver (CL) Tx. The authors report on the 1-year follow-up of their cohort of hepatitis C virus (HCV) patients undergoing AALTx or CLTx. METHODS: Twenty-six patients with HCV end stage liver cirrhosis underwent CLTx and 17 underwent AALTx. The diagnosis of recurrent HCV was made on the basis of increased transaminases, detectable HCV RNA levels, and histologic findings on liver biopsy. Liver biopsies were performed on the basis of clinical indications. Bilirubin concentration, partial thromboplastin time, and alanine aminotransferase activity were compared between the two groups at different time intervals. RESULTS: HCV recurrence was seen in 10 of 26 CLTx patients versus 6 of 17 AALTx patients (P=0.1). Time until recurrence was longer in AALTx patients (158+/-114 days vs. 227+/-154 days, P=0.4). Of the biochemical parameters, only bilirubin concentration at week 4 was significantly different between AALTx and CLTx patients (3.1+/-4.3 mg/dL vs. 1.26+/-0.83 mg/dL, P=0.04). Overall survival and the number of patients needing retransplantation were similar in both groups. CONCLUSIONS: At a follow-up period of 1 year, there is no difference in outcome between end-stage HCV patients undergoing AALTx or CLTx. PMID- 14742984 TI - Mycophenolate mofetil reduces deterioration of renal function in patients with chronic allograft nephropathy. A follow-up study by the Spanish Cooperative Study Group of Chronic Allograft Nephropathy. AB - BACKGROUND: Although studies have shown that mycophenolate mofetil (MMF) with cyclosporine (CsA) and prednisone can reduce the incidence of acute rejection and increase the half-life of the graft, the effects of MMF on established chronic allograft nephropathy (CAN) are controversial. METHODS: We studied 121 patients with biopsy-proven CAN, 59 treated with CsA and prednisone and 62 treated with triple-drug therapy with azathioprine. At inclusion, each group received 2 g per day of MMF and azathioprine was stopped. Renal function was measured by the glomerular filtration rate (GFR) obtained by creatinine clearance (Cockcroft Gault) and monitored by the slope of the GFR, adjusted using linear regression. RESULTS: The median follow-up, after incorporation of MMF, was 36 (13-36) months, with 103 (85.1%) having a full 3-year follow-up. Before the introduction of MMF, there was progressive deterioration in renal function (GFR: 54.8+/-20.9 vs. 39.7+/-14.0 mL/min, P<0.001). After introduction of MMF, renal function remained stable (GFR: 39.7+/-14.0 vs. 41.3+/-10.8 mL/min, P=NS), with a significant change in the slope of the GFR (-0.0144 vs. +0.00045, P<0.001). In 65 patients in whom CsA blood levels remained unchanged during follow-up (148.0+/-65.6 vs. 154.1+/ 58.2, P=NS), the slope of the GFR showed a reduction in loss of renal function ( 0.0147 vs. -0.0001, P<0.001). CONCLUSIONS: Treatment with MMF reduced the progressive deterioration of renal function in patients with CAN, independently of the blood levels of CsA. PMID- 14742985 TI - Relationship between postrenal transplant osteonecrosis of the femoral head and gene polymorphisms related to the coagulation and fibrinolytic systems in Japanese subjects. AB - BACKGROUND: Nontraumatic osteonecrosis of the femoral head (ONFH) is one of the complications that may occur after renal transplantation. We investigated the relationship between the incidence of ONFH and polymorphisms in the genes for plasminogen activator inhibitor (PAI)-1, which is one of the major regulatory proteins of the fibrinolytic system, and 5,10-methylenetetrahydrofolate reductase (MTHFR), which is associated with the plasma levels of homocysteine in Japanese subjects. METHODS: Thirty-one patients with postrenal transplant ONFH and 106 patients without ONFH were selected. Genotypes of PAI-1 4G/5G and MTHFR C677T were determined by direct sequencing of genomic DNA. In addition, plasma PAI-1 antigen (Ag) levels and plasma total homocysteine (tHcy) levels at the steady state were measured. The relationships between the incidence of ONFH and these genotypes, as well as plasma levels of the gene products, were investigated. RESULTS: Plasma PAI-1 Ag levels were the highest in patients with the 4G/4G genotype, and plasma tHcy levels were the highest in patients with TT genotypes of MTHFR C677T. However, the relationship between the incidence of ONFHH and PAI 1 4G/5G or MTHFR C677T was not observed. The relationship between the incidence of ONFH and plasma levels of PAI-1 Ag or tHcy was not observed. CONCLUSIONS: Genotypes of PAI-1 4G/5G and MTHFR C677T or plasma concentrations of PAI-1 Ag and tHcy had no effect on the incidence of ONFH in Japanese subjects, unlike the results of studies performed in white subjects. The effect of genetic background on the pathologic conditions that developed in patients with postrenal transplant ONFH may differ according to race. PMID- 14742986 TI - Long-term outcome of liver transplants for chronic hepatitis C: a 10-year follow up. AB - BACKGROUND: Recurrence of hepatitis C (HCV) infection after orthotopic liver transplantation (OLT) in HCV-positive patients is almost universal. Severity of graft hepatitis increases during the long-term follow-up, and up to 30% of patients develop severe graft hepatitis and cirrhosis. However, there are still no clear predictors for severe recurrence. The aim of this study was to examine the 10-year outcome and risk factors for graft failure caused by HCV recurrence. METHODS: In a prospective analysis, 234 OLTs in 209 HCV-positive patients with a median age of 53 years were analyzed. Immunosuppression was based on cyclosporine A or tacrolimus in different protocols. Predictors for outcome were genotype, viremia, donor variables, recipient demographics, postoperative immunosuppression, and human leukocyte antigen (HLA) compatibilities. RESULTS: Actuarial 5-, and 10-year patient survival was 75.8% and 68.8%. Eighteen of 209 (8.7%) patients died because of HCV recurrence, which was responsible for 35.9% of the total 53 deaths. Significant risk factors for HCV-related graft failure in an univariate analysis were multiple steroid pulses, use of OKT3, and donor age greater than 40. However, in a multivariate analysis, multiple rejection treatments with steroids and OKT3 treatment proved to be significantly associated with HCV-related graft loss. CONCLUSIONS: The analysis of causes leading to graft failure in patients with HCV showed that HCV recurrence is responsible for one of three deaths in HCV-positive patients. Rejection treatment contributed significantly to an enhanced risk for HCV-related graft loss. New antiviral treatments, as well as adapted immunosuppressive protocols, will be necessary to further improve the outcome of HCV-positive patients after liver transplantation. PMID- 14742987 TI - Comparative safety of amphotericin B lipid complex and amphotericin B deoxycholate as aerosolized antifungal prophylaxis in lung-transplant recipients. AB - BACKGROUND: Aerosolized administrations of amphotericin B deoxycholate (AmBd) and amphotericin B lipid complex (ABLC) in lung transplant recipients were compared for safety and tolerability. The incidence of invasive fungal infections in patients receiving aerosolized amphotericin B formulations as sole prophylaxis was determined. METHODS: A prospective, randomized (1:1), double-blinded trial was conducted with 100 subjects. AmBd and ABLC were administered postoperatively by nebulizer at doses of 25 mg and 50 mg, respectively, which were doubled in mechanically ventilated patients. The planned treatment was once every day for 4 days, then once per week for 7 weeks. Treatment-related adverse events and invasive fungal infections were quantitated for 2 months after study drug initiation. RESULTS: Intent-to-treat analysis revealed study drug was discontinued for intolerance in 6 of 49 (12.2%) and 3 of 51 (5.9%) patients in the AmBd- and ABLC-treated groups, respectively (p=0.313). Subjects receiving AmBd were more likely to have experienced an adverse event (odds ratio 2.16, 95% confidence interval 1.10, 4.24, p=0.02). Primary prophylaxis failure within 2 months of study drug initiation was observed in 7 of 49 (14.3%) AmBd-treated patients and 6 of 51 (11.8%) ABLC-treated patients. No fungal pneumonias were observed. Only two (2%) patients experienced documented primary prophylaxis failure with Aspergillus infections within the follow-up period. CONCLUSIONS: Both aerosol AmBd and ABLC appear to be associated with a low rate of invasive pulmonary fungal infection in the early posttransplant period. Patients receiving ABLC were less likely to experience a treatment-related adverse event. PMID- 14742988 TI - Hemophagocytic syndrome in renal transplant recipients: report of 17 cases and review of literature. AB - BACKGROUND: Hemophagocytic syndrome (HPS) combines febrile hepatosplenomegaly, pancytopenia, hypofibrinemia, and liver dysfunction. It is defined by bone marrow and organ infiltration by activated, nonmalignant macrophages phagocytizing blood cells. HPS is often caused by an infectious or neoplastic disease and has rarely been described in renal transplant recipients. METHODS: We retrospectively analyzed 17 cases of HPS after cadaveric renal transplantation (13 men and 4 women, age 41+/-8 years). The median time between transplantation and hemophagocytosis was 52 days. Eleven patients (64%) had received antilymphocyte globulins during the 3 months before presentation. RESULTS: Fever was present in all patients, and hepatosplenomegaly was present in 9 of 17 patients. Other nonspecific clinical findings included abdominal, neurologic, and respiratory symptoms. Laboratory tests showed anemia (hemoglobin 6.1+/-1.3 g/dL), thrombocytopenia (34,000+/-32,000/mm3), and leukopenia (1,700+/-1,400/mm3). Elevated liver enzymes were present in 12 of 17 patients, and cholestasis was present in 10 of 17 patients. Elevated triglycerides and ferritin were noted in 75% and 86% of cases, respectively. HPS was related to viral infection in nine patients (cytomegalovirus, Epstein-Barr virus, human herpesvirus 6, and human herpesvirus 8), bacterial infection in three patients (tuberculosis and Bartonella henselae), and other infections in two patients (toxoplasmosis and Pneumocystis carinii pneumoniae). Posttransplant lymphoproliferative disease was present in two patients. Despite large-spectrum anti-infectious treatment and dramatic tapering of immunosuppression, death occurred in eight patients (47%). Graft nephrectomy was performed in four of the nine surviving patients. CONCLUSIONS: We report here the largest series of HPS after renal transplantation. This rare disease is usually secondary to herpes viridae infections, mostly cytomegalovirus and Epstein-Barr virus in severely immunocompromised patients. Despite aggressive treatment, the prognosis remains poor. PMID- 14742989 TI - A randomized long-term trial of tacrolimus and sirolimus versus tacrolimus and mycophenolate mofetil versus cyclosporine (NEORAL) and sirolimus in renal transplantation. I. Drug interactions and rejection at one year. AB - BACKGROUND: To reduce long-term nephrotoxic calcineurin inhibitor dosage, adjunctive sirolimus or mycophenolate mofetil (MMF) was used in a 150-patient, randomized, three-armed trial in cadaveric or human leukocyte antigen non identical living-donor first renal transplant recipients (n=50/group). METHODS: Group A received tacrolimus and sirolimus. Target tacrolimus trough levels postoperatively were 10, 8, and 6 ng/mL at 1 month, 6 months, and 1 year, respectively. Group B received tacrolimus and MMF. Target tacrolimus trough levels were 10 and 8 ng/mL at 1 month and 1 year, with a targeted dose of MMF of 1 g twice daily. Group C received cyclosporine A (CsA) (Neoral, Novartis, Basel, Switzerland) and sirolimus with target CsA trough levels of 225 and 175 ng/mL at 1 month and 1 year. Maintenance sirolimus target trough levels were 8 ng/mL in groups A and C. Each group received daclizumab induction and methylprednisolone maintenance. This first of two companion 1-year reports details demographics, drug-dosing interactions, and rejection. RESULTS: There were no notable differences in group demographics, but a somewhat less favorable course occurred in group C, despite higher bioavailability of sirolimus in group C versus group A (P<0.001). Acute rejection rates were lower in groups A (4%) and B (4%) combined versus group C (14%) (P=0.03). Histopathologic findings were supported by comparing perioperative with 1-year postoperative protocol biopsies. CONCLUSIONS: This 1-year interim analysis indicates that a decreasing dosage of tacrolimus with either adjunctive sirolimus or MMF may optimize future graft survival versus a less favorable outcome using a similar algorithm with CsA and sirolimus. PMID- 14742990 TI - A randomized long-term trial of tacrolimus/sirolimus versus tacrolimus/mycophenolate mofetil versus cyclosporine (NEORAL)/sirolimus in renal transplantation. II. Survival, function, and protocol compliance at 1 year. AB - BACKGROUND: In an attempt to reduce chronic calcineurin inhibitor induced allograft nephropathy in first cadaver and human leukocyte antigen non-identical living-donor renal transplantation, sirolimus (Siro) or mycophenolate mofetil (MMF) was tested as adjunctive therapy, with planned dose reductions of tacrolimus (Tacro) over the first year postoperatively. Adjunctive Siro therapy with a similar dose reduction algorithm for Neoral (Neo) was included for comparison. METHODS: The detailed dose reduction plan (Tacro and Siro, group A; Tacro and MMF, group B; Neo and Siro, group C) is described in our companion report in this issue of Transplantation. The present report documents function, patient and graft survival, protocol compliance, and adverse events. RESULTS: As mentioned (in companion report), group demographics were similar. The present study shows no significant differences in 1-year patient and graft survival but does show a trend that points to more difficulties in group C by way of a rising slope of serum creatinine concentration (P=0.02) and decreasing creatinine clearance (P=0.04). There were more patients who discontinued the protocol plan in group C. Thus far, no posttransplant lymphomas have appeared, and infectious complications have not differed among the groups. However, a greater percentage of patients in group C were placed on antihyperlipidemia therapy, with an (unexpected) trend toward a higher incidence of posttransplant diabetes mellitus in this group. Group A required fewer, and group B the fewest, antihyperlipidemia therapeutic interventions (P<0.00001). CONCLUSIONS: This 1-year interim analysis of a long-term, prospective, randomized renal-transplant study indicates that decreasing maintenance dosage of Tacro with adjunctive Siro or MMF appears to point to improved long-term function, with reasonably few adverse events. PMID- 14742991 TI - Calcineurin phosphatase activity: activation by glucocorticoids and role of intracellular calcium. AB - BACKGROUND: Glucocorticoids stimulate release of intracellular calcium in peripheral lymphocytes, but their effects on calcineurin phosphatase activity are unknown. METHODS: Calcineurin phosphatase activity was measured in permeabilized Jurkat T cells using a specific orthophosphate substrate. Changes in intracellular calcium were measured by FURA-2 fluorescence. Inositol triphosphate levels were measured by radioimmunoassay. Transfection with luciferase reporter plasmids linked to glucocorticoid response elements were used to evaluate glucocorticoid receptor function in Jurkat T cells. RESULTS: Dexamethasone significantly (P<0.004) increased calcineurin activity within 15 sec, peaking at 10 min (P<0.001) and returning to basal levels by 180 min. Inhibition of DNA transcription with actinomycin D failed to block calcineurin activation, but co incubation with RU-486 completely blocked enzyme stimulation. To determine whether Jurkat T cells express active glucocorticoid receptors, cells were transfected with a luciferase reporter plasmid linked to a glucocorticoid response element (GRE). Jurkat T cells incubated with dexamethasone (10 microM) for 24 hr failed to stimulate luciferase activity, whereas cells co-transfected with a transcriptionally active glucocorticoid receptor resulted in a doubling of luciferase activity. Dexamethasone rapidly increases intracellular inositol triphosphate (IP3) and intracellular calcium within 15 sec. Cells incubated with U-73122 (a nonspecific phospholipase C [PLC] antagonist) completely blocked dexamethasone-induced activation of calcineurin, whereas U-73343 failed to block enzyme activation. Dexamethasone-induced activation of calcineurin activity stimulates dephosphorylation of the proapoptotic protein BAD and augments apoptosis through a calcineurin-dependent pathway. CONCLUSION: Dexamethasone rapidly increases calcineurin activity through a transcription-independent mechanism involving activation of phospholipase C and the release of IP3 dependent calcium stores. PMID- 14742992 TI - Allogeneic reaction induces dendritic cell maturation through proinflammatory cytokine secretion. AB - BACKGROUND: A bone marrow transplantation conditioning regimen is known to activate host dendritic cells (DC), which then become able to initiate graft versus-host disease (GVHD) by presenting alloantigens. In this article, the authors addressed whether the alloreaction could reciprocally maintain DC in an activation state through secretion of proinflammatory cytokines. METHODS: Skin biopsy specimens from GVHD patients were analyzed for the presence of DC. Supernatants collected from primary major histocompatibility antigen (allogeneic mixed leukocyte reaction [MLR] supernatant [SN]) or secondary minor histocompatibility antigen-mismatched mixed lymphocyte reactions were used to culture cytokine-promoted immature (im) DC. DC phenotype, function, and migration were analyzed. RESULTS: Immunostaining from GVHD skin biopsy specimens showed a deficit of Langerhans cells (LC) in the epidermis but the presence of mature DC in the dermis. Because LC should have recovered in the epidermis by this time, the authors then addressed whether the allogeneic reaction could maintain DC in an activation and migratory state, through secretion of inflammatory cytokines. With this aim, cytokine-mediated imDC were exposed to alloMLR-SN for 2 days. The authors observed that DC increased their expression of CD80, CD86, CD40, and human leukocyte antigen (HLA)-DR and neoexpressed CD83, DC-LAMP/CD208, and CCR7. At the functional level, alloMLR-SN-treated DC lost their ability to capture dextran, improved their allostimulatory capacity, and migrated in response to macrophage inflammatory protein 3beta. Interestingly, SN collected from secondary HLA-identical but minor histocompatibility antigen-mismatched MLR induced almost equivalent DC phenotypic maturation. CONCLUSIONS: The authors' results show that the allogeneic reaction leads to maturation and migration of DC through proinflammatory cytokine secretion. This might contribute to the impairment of LC reconstitution in the skin of patients with GVHD. PMID- 14742993 TI - Everolimus and mycophenolate mofetil are potent inhibitors of fibroblast proliferation after lung transplantation. AB - BACKGROUND: Dysregulated fibroblast proliferation is thought to play an important role in the progression of bronchiolitis obliterans (BO) after lung transplantation. Augmented immunosuppression is often used to treat BO. We investigated the effect of methylprednisolone (mPRED), cyclosporine A (CsA), tacrolimus (FK506), azathioprine (AZA), mycophenolate mofetil (MMF), and everolimus (rapamycin derivative [RAD]) on the proliferative capacity of fibroblasts cultured from transbronchial biopsies of lung transplant recipients. METHODS: Primary cultures of human lung fibroblasts were obtained from 14 transbronchial biopsies of lung transplant recipients. Subconfluent cells were serum starved for 24 hr followed by growth stimulation in the presence or absence of the respective drug in six concentrations ranging as follows: 0.01 to 100 mg/L for mPRED; 0.01 to 50 mg/L for CsA and AZA; 0.001 to 5 mg/L for FK506 and MMF; and 0.00001 to 1 mg/L for RAD. Proliferation was quantified by [3H]thymidine incorporation and direct cell count. A toxic drug effect was excluded by trypan blue. RESULTS: Drug concentrations (mg/L) causing a 50% inhibition of fibroblast proliferation were mPRED 4; CsA 20; FK506 0.3; AZA 7; MMF 0.3; and RAD 0.0006. Drug concentrations (mg/L) causing inhibition of fetal bovine serum-induced proliferation were mPRED 60; CsA 45; FK506 3; AZA 35; MMF 1; and RAD 0.003. CONCLUSIONS: RAD and MMF were the most potent antifibroproliferative drugs and were effective at concentrations achieved clinically, supporting their use for the treatment of patients with early BO. Our method holds promise as an in vitro model to assess the likely in vivo responses of human lung fibroblasts to specific immunosuppressive drugs. PMID- 14742994 TI - The fractional excretion of soluble interleukin-2 receptor-alpha is an excellent predictor of the interleukin-2 receptor-alpha status after treatment with daclizumab. AB - BACKGROUND: Daclizumab is a humanized monoclonal antibody against the alpha-chain of the interleukin (IL)-2 receptor (R). The authors previously have shown that the urinary excretion of soluble (s) IL-2Ralpha is dependent on the presence of daclizumab in serum. The authors investigated whether the IL-2Ralpha status, as assessed by flow cytometric analysis, is reflected by the concentration of sIL 2Ralpha in the urine and serum. METHODS: Two hundred seventy-two measurements were performed in 46 renal transplant recipients who were treated with daclizumab in combination with tacrolimus and mycophenolate mofetil. Soluble IL-2Ralpha was measured in urine and serum with Immulite IL-2R, a solid-phase enzyme-linked immunosorbent assay. Complete blockade of the IL-2Ralpha was defined as the presence of less than 5% IL-2Ralpha+ lymphocytes in the CD3+ population. Receiver operating characteristic (ROC) curve analysis was performed to evaluate the performance of serum and urine sIL-2Ralpha in predicting IL-2Ralpha blockade. RESULTS: The calculated fractional excretion of sIL-2Ralpha proved to be an excellent predictor of the blockade of IL-2Ralpha (ROC analysis area under the curve, 0.95+/-0.01). A calculated fractional excretion of sIL-2Ralpha lower than 0.5% had a specificity of 100% and a sensitivity of 75% for the assessment of blockade of IL-2Ralpha. CONCLUSIONS: Blockade of IL-2Ralpha after treatment with daclizumab can reliably be assessed by calculation of the fractional excretion of sIL-2Ralpha. This method is easier to use compared with flow cytometric analysis of IL-2Ralpha+ lymphocytes. PMID- 14742995 TI - Effects of adding P38 mitogen-activated protein-kinase inhibitor to celsior solution in canine heart transplantation from non-heart-beating donors. AB - BACKGROUND: The activation of p38 mitogen-activated protein kinase (MAPK) plays an important role in ischemia-reperfusion injury. This study evaluated the effects of p38 MAPK inhibition using FR167653, a novel p38 MAPK inhibitor, as an additive to Celsior solution in canine heart transplantation from non-heart beating donors (NHBDs). METHODS: Donor hearts were left in situ for 20 minutes after cardiac arrest, which was induced by rapid exsanguination. Twelve donor recipient pairs of mongrel dogs were divided into two groups: the control and FR167653 (FR) groups (n=6 each). In both groups, the grafts were subjected to coronary flushing and immersed in Celsior solution for 4 hours with or without FR167653. Orthotopic heart transplantation was then performed. Cardiac output (CO), left ventricular pressure (LVP), and end-systolic maximal elastance (Emax) were measured 2 hours after weaning from cardiopulmonary bypass (CPB), and the hearts were then harvested for histopathologic study. The activation of p38 MAPK was evaluated in another 20 mongrel dogs. RESULTS: In the FR group, CO, LVP recovery rate, and Emax were significantly (P<0.05) higher 2 hours after weaning from CPB, histopathologic damage was attenuated, and the activation of p38 MAPK was significantly (P<0.05) inhibited 10 minutes after reperfusion compared with the control group. CONCLUSIONS: The addition of FR167653 to Celsior solution improved heart-graft viability, probably by way of the inhibition of p38 MAPK activation, which may attenuate ischemia-reperfusion injury in heart transplantation from NHBDs. PMID- 14742996 TI - Effect of beta2-adrenergic receptor agonists on intercellular adhesion molecule (ICAM)-1, B7, and CD40 expression in mixed lymphocyte reaction. AB - BACKGROUND: The plasma interleukin (IL)-18 level is elevated in acute rejection after organ transplantation. Although beta2-adrenergic receptor (AR) agonists suppress the rejection of organ and tissue transplants, little is known about their action mechanisms. We examined the effects of endogenous catecholamines and beta2-AR agonists on the expression of intercellular adhesion molecule (ICAM)-1, B7.1, B7.2, CD40, and CD40 ligand (CD40L) in human mixed lymphocyte reaction (MLR) and in an in vitro model of acute rejection in the presence or absence of IL-18. METHODS: ICAM-1, B7.1 B7.2, CD40, and CD40L expression on monocytes was measured by flow cytometry, and the production of interferon (IFN)-gamma and IL 12 was determined by enzyme-linked immunosorbent assay. Lymphocytes proliferation in MLR was measured by [3H]-thymidine uptake. The relevant AR subtypes were characterized using subtype-selective agonists and antagonists. RESULTS: beta2-AR agonists inhibited the expression of ICAM-1 and CD40 during MLR in the absence of IL-18. Among IL-18-induced expression of ICAM-1, B7.1, B7.2, CD40, and CD40L, beta2-AR agonists inhibited ICAM-1 and CD40 expression. beta2-AR agonists prevented the production of IFN-gamma and IL-12 in the presence of IL-18 but had no effect in the absence of IL-18. beta2-AR agonists inhibited lymphocyte proliferation in IL-18-treated MLR. CONCLUSIONS: We found that beta2-AR agonists strongly inhibited the expression of ICAM-1 and CD40, irrespective of the presence or absence of IL-18, which is different from that of histamine and prostaglandin E2. PMID- 14742997 TI - Improved survival of orthotopic liver allograft in swine by addition of trophic factors to University of Wisconsin solution. AB - Serum-free preservation media such as University of Wisconsin (UW) may cause tissue damage through trophic factor (TF) deprivation. This study evaluated whether the addition of TFs to UW solution improves liver graft quality after extended cold preservation time in pigs. UW solution was supplemented with epidermal growth factor, insulin-like growth factor-1, nerve growth factor-beta, bactenecin, and substance P to create TF-supplemented (TFS) UW. Orthotopic liver transplantation was performed after 18 hr of static cold storage at 4 degrees C in UW (n=7) or TFS-UW (n=7) solution. Recipients of grafts preserved with TFS-UW demonstrated significantly better 5-day survival (57%) than those preserved with UW alone (14%) (P<0.05). Adenosine triphosphate content in grafts preserved in TFS-UW was significantly higher than in grafts preserved in UW (17.4+/-5.0 vs. 4.8+/-1.2 nmol/mg protein, respectively) (P<0.05). This study showed that the addition of TFs to UW solution allowed a significant extension of cold ischemic time in pigs. PMID- 14742998 TI - Randomized controlled trial of sequential intravenous and oral ganciclovir versus prolonged intravenous ganciclovir for long-term prophylaxis of cytomegalovirus disease in high-risk cytomegalovirus-seronegative liver transplant recipients with cytomegalovirus-seropositive donors. AB - Cytomegalovirus (CMV)-seronegative liver transplant recipients with CMV seropositive donors have the greatest risk for CMV disease. We performed a randomized, controlled trial comparing sequential intravenous (IV) and oral ganciclovir with prolonged IV ganciclovir for long-term prophylaxis of CMV disease in these high-risk patients. Patients were initially given IV ganciclovir at a dose of 6 mg/kg per day from days 1 to 14 after transplantation. Patients then either received oral ganciclovir (1 g every 8 hr) or continued IV ganciclovir (6 mg/kg once per day on Monday-Friday of each week) from days 15 to 100 after transplantation. CMV disease occurred in 3 of 32 patients (9.3%) receiving oral ganciclovir and in 4 of 32 patients (12.5%) receiving IV ganciclovir within the first year after transplantation (P>0.2). All cases of CMV disease occurred more than 90 days after transplantation (median time of onset day +137 for oral ganciclovir and day +135 for IV ganciclovir). There were no deaths from CMV in either study group. Both oral and IV ganciclovir were generally well tolerated. These results indicate that, after induction with 14 days of IV ganciclovir, oral ganciclovir can be as effective as IV ganciclovir for long-term prophylaxis of CMV disease in high-risk CMV-seronegative liver transplant recipients with CMV-seropositive donors and eliminates the need for prolonged IV access. PMID- 14742999 TI - Insulin independence achieved using the transmesenteric approach to the portal vein for islet transplantation. AB - Clinical human islet transplantation has been performed successfully using a percutaneous transhepatic approach to access the portal vein. The risks from percutaneous delivery of islets, such as bleeding and puncturing neighboring structures, can be avoided by a transmesenteric approach to the portal vein, which we have used to stent completely or near-completely occluded portal veins in both cirrhotic and noncirrhotic patients with minimum morbidity. After minilaparotomy, a second-order tributary branch of the mesenteric vein is cannulated to provide endovascular access to the portal vein. The islet preparation is infused through a catheter directed under fluoroscopy from the mesenteric vein to the portal vein. Pre- and postinfusion portograms are obtained to confirm the absence of any interval changes in portal venous flow. We have performed this procedure successfully in three islet-transplant recipients each receiving two infusions on separate occasions, with some of these procedures performed under local anesthesia without complications. The transmesenteric approach promises to be a safe alternative to percutaneous islet delivery. PMID- 14743000 TI - Increased incidence of acute myeloid leukemia after liver transplantation? Description of three new cases and review of the literature. AB - BACKGROUND: Acute myeloid leukemia (AML) after solid organ transplantation is rare, with only six AML cases after liver transplantation (LT) being reported. METHODS: Characteristics of three AML cases observed among 799 recipients of LT in the authors' institution and estimation of the standardized incidence ratio (SIR) are presented. RESULTS: Three AML cases were diagnosed (French-American British classification subtypes M0, M3, and M1) at 38 days, 2 years, and 3.5 years after LT, respectively. The immunosuppressive regimen consisted of cyclosporine A and prednisone. The recipient's origin of blasts could be demonstrated in two cases. All patients achieved complete remission after standard treatment. Nonetheless, disease relapsed in two patients at 2 and 6 months, whereas the patient with acute promyelocytic leukemia remains disease free 4 years after diagnosis. As compared with the general population, the observed incidence of AML in the authors' series of LT results in a significantly higher SIR of 11.41 (P=0.0023). CONCLUSIONS: The higher SIR found in the authors' series suggests an increased risk of AML in patients undergoing LT. PMID- 14743001 TI - The role of complement receptors in production of antibodies specific for Galalpha1,3Gal. AB - By using alpha-galactosyl transferase knockout (GT-/-) mice, which make natural alphaGal-reactive antibodies, we examined the role of complement receptors in the production of alphaGal-specific antibodies. GT-/- mice were crossed with complement receptor 2 loci knockout mice to generate double knockout (DKO) mice. alphaGal-specific natural antibodies were detectable by enzyme-linked immunosorbent assay in the serum of GT-/- mice by 9 weeks of age. In contrast, only low titers of alphaGal-specific natural antibodies were detectable only in the serum of older DKO mice. Serum titers of alphaGal-reactive antibodies in GT-/ mice increased significantly after immunization with pig cells. In contrast, immunization had little effect on alphaGal-reactive antibody levels in DKO mice. Similarly, pretreatment of GT-/- mice with a blocking antibody to CD21 and CD35 inhibited production of alphaGal-reactive antibodies after immunization. However, DKO mice were able to make alphaGal-specific antibodies after secondary immunization. Thus, Cr2 loci-encoded receptors seem to be directly involved in the production of primary alphaGal-reactive antibodies. PMID- 14743002 TI - Absence of graft-versus-host disease in the isolated vascularized bone marrow transplant. AB - An isolated vascularized bone marrow transplant (iVBMT) model was developed to study the contribution of the bone marrow component in a composite tissue allograft. We hypothesized that the iVBMT would be functional and cause graft versus-host disease (GVHD) in a fraction of the recipients. Lewis iVBMT grafts were transplanted to Lewis-Brown Norway recipients. Animals were sacrificed at various times from 1 to 14 weeks. Polymerase chain reaction for microchimerism was performed on the host's marrow. No animals exhibited signs of GVHD at death. Histologic examination of the grafts showed a normal mix of hematopoietic and fatty elements and appeared to be functional. Tissues usually affected-tongue, ear, liver, and gut-also showed no evidence of disease. Polymerase chain reaction demonstrated microchimerism in both groups. These findings suggest that the vascularized bone marrow within a composite tissue allograft is not the component that causes GVHD; rather, it may serve an immunomodulatory function for tolerance induction. PMID- 14743003 TI - Parvovirus B19 infection-associated red-cell aplasia in renal-transplant recipients: clues from the bone marrow. PMID- 14743004 TI - Diabetic muscle infarction in renal transplantation. PMID- 14743005 TI - Rapid intravenous desensitization to antithymocyte globulin in a patient with aplastic anemia. PMID- 14743006 TI - Patient management by cyclosporine C2 monitoring. PMID- 14743007 TI - Delayed domino liver transplantation: use of the remnant liver of a recipient of a temporary auxiliary orthotopic liver transplant as a liver graft for another patient. PMID- 14743008 TI - Ultrastructural demonstration of replicative herpes simplex virus type 1 transmission through corneal graft. PMID- 14743009 TI - Portal biliopathy treated by liver transplantation. PMID- 14743010 TI - Role of cytomegalovirus infection in chronic allograft nephropathy. PMID- 14743011 TI - Difficult lens power calculations. AB - PURPOSE OF REVIEW: Although cataract extraction seems to be feasible without major technical obstacles, the surgical technique has changed completely, and patients are no longer satisfied with good spectacle-corrected vision but anticipate complete visual rehabilitation after cataract surgery, without correction. To fulfill this desire, toric or accommodative intraocular lenses are of increasing popularity, and the intraocular lens power calculation after keratorefractive surgery has been improved. RECENT FINDINGS: In this review article, we provide an overview of different mathematical strategies of calculating the intraocular lens power with standard formulas and with new algorithms, such as paraxial or numeric ray-tracing. These enhanced techniques may improve the validity of lens power calculation due to reduction of the prediction error, especially in cases with high or excessive corneal astigmatism and after refractive laser surgery. Furthermore, a new calculation scheme for the determination of bitoric eikonic intraocular lenses allows a distortion-free imaging in astigmatic eyes. The biometric determinants for the different formulas and calculation schemes are discussed in detail. SUMMARY: In difficult cases, standard calculation schemes are overemployed and new mathematical algorithms are necessary to adequately address these problems. Ray-tracing algorithms and other complex mathematical computation schemes are of increasing interest and will more and more replace conventional calculation formulas for determination of intraocular lens power. PMID- 14743013 TI - Genetics of childhood cataract. AB - PURPOSE OF REVIEW: Congenital cataracts, although much less common than their age related counterparts, account for one-tenth of cases of childhood blindness. Approximately half are inherited, either in isolation or as part of a syndrome of ocular or systemic anomalies. This article reviews recent advances made in understanding the molecular genetic basis of isolated, nonsyndromic inherited cataract. RECENT FINDINGS: New disease-causing mutations continue to be identified and now encompass genes encoding a wide variety of different lens proteins. More detailed investigations of the functional consequences of each mutation are being reported and suggest that lens opacification results not only from precipitation and amyloid-like accumulation of proteins essential for lens transparency but also from interference with their secondary functions. SUMMARY: Improved functional characterization of mutations causing childhood cataract will improve understanding of lens development and physiology but will also have implications for the more common age-related cataract. This too has a significant genetic component to its etiology, and genes causing monogenic forms of childhood inherited cataract represent excellent candidate genes for age-related cataract. The identification of the genes conferring increased risk of developing age related cataract will bring closer the development of a medical treatment to delay the onset of lens opacification and need for surgery. PMID- 14743014 TI - Accommodating intraocular lenses. AB - PURPOSE OF REVIEW: With the advent of interest in accommodative lenses as a solution for presbyopia and the growing baby-boomer demographic, ophthalmic surgeons will have the opportunity to provide this technology to facilitate near, intermediate, and distance vision for their patients. RECENT FINDINGS: At present, six corporate entities and lens designs are attempting to commercialize accommodative intraocular lens devices. One Food and Drug Administration (FDA) clinical trial has been completed and the first FDA-approved accommodating lens is available. SUMMARY: For the first time, ophthalmic surgeons will be able to provide a full range of visual focus in each eye of a patient to maintain binocular function while also avoiding the unwanted mesopic and scotopic visual disturbances that are experienced with monovision and multifocal lens technologies. Accommodative intraocular lenses could revolutionize not only cataract visual rehabilitation but also the surgical approach to presbyopia. PMID- 14743015 TI - Refractive lens exchange as a refractive surgery modality. AB - PURPOSE OF REVIEW: Refractive lens exchange 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. A review of the recent literature was performed to determine recent advances in this surgical procedure. RECENT FINDINGS: New multifocal and accommodative lens technology should enhance patient satisfaction. In addition, newer lens extraction techniques using microincisions and new phacoemulsification technology will enhance the safety of this procedure, ultimately allowing refractive lens exchanges to be performed through two microincisions as future lens technologies become available. SUMMARY: Attention to detail in regard to proper patient selection, preoperative measurements, intraoperative technique, and postoperative management has resulted in excellent outcomes and improved patient acceptance of this effective technique. PMID- 14743016 TI - Survey of ophthalmic viscosurgical devices. AB - PURPOSE OF REVIEW: Cataract treatment is evolving, thanks to new phaco systems and innovative surgical techniques. Recent ophthalmic viscosurgical devices (OVDs) play an important role in the development of new approaches to surgery. Knowledge of the new techniques and advances in new OVDs is crucial to performing routine and complicated surgeries. RECENT FINDINGS: Viscoadaptive substances have been introduced as a particular kind of OVD and are distinguished by high viscosity at a low flow rate and a tendency to fracture at higher flow rates, assuming pseudodispersive behavior. The ultimate soft shell technique emphasizes the rheology of viscoadaptive OVDs to perform phacoemulsification procedures, exploiting all the useful properties of these OVDs in terms of space maintenance and tissue protection. Recent studies have highlighted some new and interesting aspects of the use of OVDs. Current reports suggest that the association of lidocaine and OVD seems to be safe and useful in providing anesthesia during cataract surgery. Concern has been expressed in other studies regarding the presence of contaminants, such as endotoxins, which may be responsible for postoperative uveitis-like reactions. Finally, OVDs are widely known to protect the corneal endothelium during cataract surgery. Their role is also assuming relevance in vitreoretinal surgery. SUMMARY: New OVDs and related techniques represent an advancement in cataract surgery. PMID- 14743017 TI - Managing intraoperative complications in cataract surgery. AB - PURPOSE OF REVIEW: This article surveys the literature from 2002 and 2003 and addresses a variety of complications that may arise during cataract surgery. Modern surgical techniques and technology contribute to the decreased incidence of complications. Surgeon experience, early recognition of potential complications, and optimal handling of events lead to better outcomes. RECENT FINDINGS: Recognizing eyes preoperatively that are particularly at risk for having dysgenesis of the capsular bag, cortical-capsular adhesions, low endothelial cell reserve, dropped nucleus, and bag-lens dislocation will allow for contingency planning and appropriate tailoring of techniques. Capsular tension rings are indispensable for achieving in-the-bag implantation in the presence of weak zonules and zonulolysis. Viscolevitation to rescue sinking implants, avoidance of foldable lens complications, and proper lens placement with an open capsule are discussed. High-frequency ultrasound biomicroscopy is a useful diagnostic tool. Noninjection sub-Tenon anesthesia is not benign. Complications have been documented that include traumatic optic neuropathy. Immediate diagnosis and a trial of corticosteroids could be vision saving. Mastering topical anesthesia is a worthy goal. In some cases, faulty instruments result in complications. Routine inspection of reusable instruments, or the use of disposable instruments is warranted. Though there is no FDA-recognized use, at the time of this writing, for trypan blue dye, it has become the world standard for intraocular staining because of its safety and efficacy. Clear corneal self sealing incisions are increasingly mainstream and, when properly constructed, are extremely secure. The most significant trend for successful management of intraoperative complications this year is the increasing use of pars plana anterior vitrectomy. The ability to identify prolapsed vitreous by intracameral Kenalog staining is an invaluable tool. Surgeons depend on a team of nurses and technicians for optimal outcomes. SUMMARY: With new technology and techniques highlighted by capsular tension rings, Kenalog vitreous identification, and fourth-generation fluoroquinolones, even complicated cataract surgery can result in optimal outcomes. PMID- 14743018 TI - AquaLase: a new technology for cataract extraction. AB - PURPOSE OF REVIEW: This review describes a completely new technology for removal of the crystalline lens and compares it to currently available instruments. The AquaLase option on the Infiniti Vision System (Alcon Laboratories, Fort Worth, Texas) delivers unique advantages over conventional ultrasound phacoemulsification. RECENT FINDINGS: Incremental improvements have made ultrasonic phacoemulsification safer to the posterior capsule and less likely to create wound burn. AquaLase uses a fundamentally different technology that eliminates the risk of incision burn, is probably less likely to cause posterior capsule rupture than ultrasonic phacoemulsification needles, and is more efficient than laser-based lens removal instruments. SUMMARY: AquaLase is a new technology that offers significant advantages and is likely to find wider application as clinical experience accumulates. PMID- 14743019 TI - A review of suturing techniques for posterior chamber lenses. AB - PURPOSE OF REVIEW: To evaluate the outcome of last the 15 years' experience with the transscleral suture fixation technique of posterior chamber intraocular lens (PC IOL). RECENT FINDINGS: The implant of an anterior chamber IOL, especially the iris-claw lens, is safer and a better option than the transsclerally fixed IOL. SUMMARY: After bibliographic review of anterior chamber lenses implant results with transscleral fixation, we conclude that the number of complications is less in the iris fixation lens. PMID- 14743020 TI - Pharmacologic considerations for cataract surgery. AB - PURPOSE OF REVIEW: A variety of options exist for perioperative, intraoperative, and postoperative medications in cataract surgery. This article reviews some of the more timely literature on "controversial" subjects in this area. RECENT FINDINGS: Recent literature may support the rationale for intracameral vancomycin at the time of cataract surgery. The recently released fluoroquinolones moxifloxacin and gatifloxacin are reviewed in terms of their clinical properties. Pharmacologic adjuncts to surgery are discussed, as is the use of intravitreal triamcinolone both at the time of cataract surgery and postoperatively. SUMMARY: Recent advances in ophthalmic pharmacology should aid the practicing anterior segment surgeon. This may affect preoperative, postoperative, and even intraoperative regimens. PMID- 14743021 TI - Wavefront technology in cataract surgery. AB - PURPOSE OF REVIEW: As advances in technology allow cataract surgeons to address higher order optical aberrations, the measurement of functional vision becomes increasingly critical. Contrast sensitivity testing is assuming a prominent place in our evaluation of surgical modalities because it reflects functional vision and correlates with visual performance. The Tecnis Z9000 intraocular lens (IOL) (Pfizer, New York) is the first foldable IOL designed to correct higher order optical aberrations and represents a first step toward the integration of wavefront technology and cataract surgery. RECENT FINDINGS: Contrast sensitivity declines with age, even in the absence of ocular pathology. Wavefront science demonstrates that the youthful crystalline lens compensates for aberrations in the cornea. The aging lens loses its balance with the cornea, as both the magnitude and the sign of its spherical aberration change. Older pseudophakic patients have generally the same contrast sensitivity as their age-matched counterparts without cataract. The Tecnis Z9000 IOL (Pfizer, New York) has been designed with a modified prolate anterior surface to compensate for the spherical aberration of the cornea, thus eliminating total ocular spherical aberration. Clinical data demonstrate that this modified prolate IOL provides superior functional vision, similar to that of younger people, and hence improves visual performance when compared with conventional spherical IOLs. It appears likely that the decline in functional vision with age involves changes in the spherical aberration of the crystalline lens. SUMMARY: The integration of wave-front technology and lens-based surgery represents a step toward improving functional vision and quality of life for cataract patients. PMID- 14743022 TI - Surgical strategies in patients with combined cataract and glaucoma. AB - PURPOSE OF REVIEW: In the elderly population, the combined presence of cataract and glaucoma is a frequent condition. In this situation, several surgical options are possible: cataract surgery only and later maybe trabeculectomy, trabeculectomy only and later maybe cataract surgery, or combined cataract and glaucoma surgery. This review compares the different surgical options on the basis of their achievable postoperative intraocular pressure (IOP) level and success and complication rates. RECENT FINDINGS: The impression of better IOP regulation with trabeculectomy than with phacotrabeculectomy has been recently confirmed by an evidence-based review. Contrary to this finding, the success of deep sclerectomy or trabeculotomy does not seem to be compromised by simultaneous phacoemulsification. In eyes with previous glaucoma-filtering surgery, cataract surgery with clear corneal incision has no effect on mean IOP but increases the 3 year failure probability. For phacotrabeculectomy, moderate evidence of a beneficial effect of MMC on IOP regulation and only weak evidence for separating the incisions has been recently reported by another evidence-based review. SUMMARY: The choice of the preferred surgical method depends on the target pressure, the amount of glaucomatous damage, and the grade of visual disturbance caused by the cataract. Phacotrabeculectomy combined with mitomycin C achieves the best IOP lowering of all types of combined cataract and glaucoma surgery currently possible but is associated with potentially sight-threatening complications. In the absence of a low target pressure, phacotrabeculotomy or the combination of phacoemulsification with viscocanalostomy or deep sclerectomy may be the therapy of choice. PMID- 14743023 TI - Outcome of open reduction and internal fixation of surgical neck nonunions of the humerus. AB - OBJECTIVE: The purpose of this study was to evaluate the results of open reduction and internal fixation of nonunions of the surgical neck of the humerus. DESIGN: Retrospective review of patients who underwent operative treatment of a surgical neck of the humerus nonunion. SETTING: Shoulder and elbow service at two university centers. PATIENTS: Thirteen patients with a two-part nonunion of the surgical neck of the humerus who were treated with open reduction and internal fixation with bone graft with a minimum of 12 months' follow-up were included. Patients with avascular necrosis, posttraumatic arthritis, severe humeral head bone loss, or a nonunion of one of the two tuberosities were excluded. INTERVENTION: Open reduction and internal fixation with either a blade plate or a T-plate and autogenous bone graft. MAIN OUTCOME MEASURES: Medical records, operative reports, physical examination, and preoperative and postoperative radiographs were reviewed. Outcome was assessed using Neer's criteria for the evaluation of total shoulder arthroplasty, visual analogue pain scale, range of motion, and ability to perform activities of daily living.RESULTS The results were excellent in 11 patients, satisfactory in 1 patient, and poor in 1 patient. The 11 nonunions in patients with excellent results healed within 6 months. The patient with the poor result initially had a persistent nonunion and required revision, open reduction and internal fixation, and bone grafting. This patient healed and went on to have an excellent result at final follow-up. All fractures were healed at the time of this study. Pain scores decreased from an average of 4.2 preoperatively to 1.2 postoperatively on a 5-point pain scale. Forward elevation in the scapular plane improved from 24 degrees preoperatively to 144 degrees postoperatively. All patients but one were able to attain overhead elevation. All patients were able to perform activities of daily living, such as dressing, bathing, combing hair, and performing perineal care, at final follow up. Overall satisfaction increased from an average of 1.0 to 9.4 on a 10-point visual analogue scale. CONCLUSIONS: Open reduction and internal fixation with autogenous bone graft results in excellent outcomes even in patients >65 years old and patients with significant medical problems. This treatment method offers predictable fracture healing and has a low complication rate. PMID- 14743024 TI - Correlation of interosseous membrane tears to the level of the fibular fracture. AB - OBJECTIVES: To correlate interosseous membrane (IOM) tears of the ankle to the height of fibular fractures in operative ankle fractures. DESIGN: Prospective clinical trial. SETTING: University Level 1 trauma center. PATIENTS: All patients admitted with a closed operative ankle fracture were included. Of 93 patients originally evaluated, 73 patients had adequate MRI for evaluation. INTERVENTION: Open reduction and internal fixation of each ankle fracture was performed after preoperative MRI evaluation of the IOM. Transsyndesmotic screw fixation was performed when evidence of syndesmotic instability was shown by intraoperative stress testing. MAIN OUTCOME MEASUREMENTS: Radiographs were analyzed for fracture classification and prediction of ligamentous injuries about the ankle. MRI evaluated the IOM integrity, correlating it to the height of the fibular fracture. RESULTS: Of the 73 ankle fractures with adequate MRI evaluation, 30 had identifiable complete IOM tears on MRI. Ten of the 30 IOM tears did not correlate with the level of the fractured fibula. Seven cases had IOM tears proximal to the fibular fracture as detected by MRI. Five of these cases were Weber B type fractures, and two were Weber C type fracture patterns. Conversely, three cases of Weber C type fractures had IOM tears that remained distal to the level of the fibular fracture. CONCLUSIONS: The level of the fibular fracture does not correlate reliably with the integrity or extent of the interosseous membrane tears identified on MRI in operative ankle fractures. One cannot consistently estimate the integrity of the IOM and subsequent need for transsyndesmotic fixation based solely on the level of the fibular fracture. An intraoperative syndesmotic stress test is recommended to establish the presence or absence of syndesmotic instability. PMID- 14743025 TI - One-stage lengthening using a locked nailing technique for distal femoral shaft nonunions associated with shortening. AB - OBJECTIVE: To assess the effectiveness of a one-stage lengthening using a locked nail technique for the treatment of distal femoral shaft nonunions associated with shortening. DESIGN: Retrospective. SETTING: University hospital. PATIENTS AND METHODS: During a 6-year period, 36 distal femoral shaft nonunions associated with shortening (>1.5 cm) were treated by the one-stage lengthening technique. Indications for this technique were distal femoral shaft aseptic or quiescent infected nonunions, 1.5-5 cm shortening, and a fracture level suitable for the insertion of two distal locked screws. The surgical technique involved skeletal traction using the femoral condyle, local debridement, lengthening by or =95%, decision tree models reached cross-validated specificities of 37 and 43%, with negative predictive values of 96 and 99%, for bacteremia and Gram-negative bacteremia, respectively. Absence of a clinically or radiologically evident source of infection and previous episodes of fever and neutropenia were defined as two newly described factors associated with bacteremia. CONCLUSIONS: Based on this retrospective analysis, it appears that bacteremia can be predicted with clinically useful specificity at a high level of sensitivity, using clinical information available at presentation in pediatric cancer patients with fever and neutropenia. PMID- 14743048 TI - Molecular characterization of Streptococcus pyogenes isolates collected during periods of increased acute rheumatic fever activity in Utah. AB - BACKGROUND: Salt Lake City, Utah has seen a continuing resurgence of rheumatic fever (RF) since 1985. METHODS: emm genotyping and multilocus sequence typing of streptococcal isolates from periods of increased RF activity were performed. RESULTS: Multiple genotypes were present during 1985 and 1998, two peak years of RF activity, and in 1992, a year with reduced RF activity. emm3 and emm18.1 were present in 1985 and 1998, but not in 1992. Two other emm types, 12 and L28, were significantly elevated in 1998 (a peak RF year) over 1992 (a non-peak RF year). Allelic profiles for the emm3 and emm18.1 isolates exhibited clonality. CONCLUSIONS: During years of increased RF activity multiple emm types, including emm18.1 and emm3, were circulating in the community. During a year of decreased RF activity, emm3 and emm18.1 genotypes were absent. The clonality of the emm3 and emm18.1 types suggests that specific clones of both types are important in the resurgence of RF during these peak years. Two other genotypes, emm12 and emmL28, may also be associated with the persistence of RF in the Salt Lake City, UT area. PMID- 14743050 TI - Plague. PMID- 14743051 TI - Tuberculosis screening in immigrant children. PMID- 14743052 TI - Seasonal variation in respiratory syncytial virus epidemics in the Gambia, West Africa. AB - Respiratory syncytial virus outbreaks tend to occur seasonally and are a major cause of childhood morbidity. In The Gambia a regular pattern of outbreaks during six consecutive annual seasons was disrupted by 2 years of irregular outbreaks, followed by another 2 years of regular seasonal outbreaks. Improved understanding of the transmission dynamics of respiratory syncytial virus is essential to design and test effective interventions. PMID- 14743053 TI - Persistent hepatitis associated with chronic active Epstein-Barr virus infection. AB - A previously healthy boy developed persistent hepatitis without fever or lymphoproliferative disorder. Although serologic tests were not indicative, Epstein-Barr virus (EBV) genome and transcripts were detected from the liver tissue, and real time PCR detected extremely high levels of EBV viremia. EBV infection should be included in the differential diagnoses of hepatitis of unknown etiology, even with unremarkable serologic data. PMID- 14743054 TI - Acute neonatal suppurative parotitis: case reports and review. AB - We describe two cases of neonatal suppurative parotitis caused by Staphylococcus aureus. Only 32 cases of neonatal suppurative parotitis (72% male) have been described in the English literature in the last 35 years. Thirty-eight percent were premature babies, only 41% were febrile and the causative agent in most cases was S. aureus. Recovery was achieved in 78% of the patients with antibiotic therapy without drainage. PMID- 14743055 TI - Congenital tuberculosis presenting as progressive liver dysfunction. AB - We report a case of congenital tuberculosis with an unusual presentation as progressive liver dysfunction, in the absence of respiratory symptoms. Several uncommon features were present, including petechiae, cutaneous lesions, ascites and positive peritoneal fluid culture. PMID- 14743056 TI - Toothpick puncture injuries of the foot in children. AB - We reviewed the medical records of five children seen between January 1997 and September 2003 for toothpick puncture injuries of the foot. Failure to visualize retained toothpicks by plain radiographs delayed early removal. Patients subsequently developed recurrent foot cellulitis, complicated by foot osteomyelitis in three cases. Ultrasound, computed tomography or magnetic resonance imaging detected the toothpicks. These imaging tools should be considered for initial evaluation of these patients. PMID- 14743057 TI - Legionella pneumonia: infection during immunosuppressive therapy for idiopathic pulmonary hemosiderosis. AB - We report a case of Legionella pneumonia in a 10-year-old girl with idiopathic pulmonary hemosiderosis who was chronically immunosuppressed and had exposure to a hot tub. Prompt diagnosis with bronchoalveolar lavage and subsequent antimicrobial therapy resulted in full recovery. Legionellosis should be included in the differential diagnosis of the immunosuppressed child with respiratory illness. High risk patients should avoid exposure to hot tubs. PMID- 14743058 TI - New data on influenza vaccines in children. PMID- 14743059 TI - Predictive value of serum interleukin-6 and interleukin-12 in children with chronic hepatitis B infection treated with interferon alpha. PMID- 14743060 TI - Geotrichum candidum septicemia in a child with hepatoblastoma. PMID- 14743061 TI - Outbreak of coxsackie B5 virus meningitis in a Scout camp. PMID- 14743062 TI - Selective screening for complement deficiencies in patients with meningococcal disease. PMID- 14743063 TI - Anaerobic central nervous system infections after trauma. PMID- 14743064 TI - Ganciclovir treatment of children with congenital cytomegalovirus infection. PMID- 14743065 TI - Scapular mass in an adolescent. PMID- 14743069 TI - The emergence of Neisseria gonorrhoeae with decreased susceptibility to Azithromycin in Kansas City, Missouri, 1999 to 2000. AB - BACKGROUND AND OBJECTIVES: We describe the first cluster of persons with Neisseria gonorrhoeae with decreased susceptibility to azithromycin (AziDS; minimum inhibitory concentration >/=1.0 microg/mL) in the United States. GOAL The goal of this study was to identify risk factors for AziDS N. gonorrhoeae and to describe isolate microbiology. STUDY DESIGN: Persons with AziDS N. gonorrhoeae (cases) were identified in Kansas City, Missouri, through the Gonococcal Isolate Surveillance Project (GISP) in 1999 and expanded surveillance, January 2000 to June 2001. A case-control study using 1999 GISP participants was conducted; control subjects had azithromycin-susceptible N. gonorrhoeae. RESULTS: Thirty three persons with AziDS N. gonorrhoeae were identified. Case patients were older than control patients (median age, 33 years vs. 23 years; P <0.001). Fifty percent of cases and 13% of control subjects had a history of sex with a female commercial sex worker (odds ratio, 7.0; 95% confidence interval, 1.3-36.0); 50% of cases and 4% of control subjects met sex partners on street A (P <0.01). AziDS N. gonorrhoeae isolates were phenotypically and genotypically similar and contained an mtrR gene mutation. CONCLUSIONS: With few treatment options remaining, surveillance for antimicrobial-resistant N. gonorrhoeae is increasingly important, especially among persons at high risk. PMID- 14743070 TI - Differences between nonnational and indigenous patients with sexually transmitted infections in Italy and insight into the control of sexually transmitted infections. AB - BACKGROUND: To determine the health needs non-nationals, information on disease occurrence is fundamental. GOALS The goal of this study was to assess the clinical and behavioral characteristics of non-Italians with a new sexually transmitted infection (STI). METHODS: Data were taken from Italy's National STI Surveillance System (1991-1999). RESULTS: Of the 61,798 STI cases reported from January 1991 to December 1999, 6847 (11.2%) were diagnosed among non-Italians, 47.1% of whom were Africans. Being a non-Italian patient with an STI was associated with male gender (adjusted odds ratio [AOR], 1.19; 95% confidence interval [CI], 1.11-1.27), youngest age (AOR, 1.71; 95% CI, 1.43-2.04), no formal education (AOR, 20.25; 95% CI, 17.51-23.42), and having contracted the STI abroad (AOR, 13.98; 95% CI, 12.59-15.54). The HIV-1 prevalence among non-Italian patients with STIs was 5.5% (95% CI, 4.9-6.2), with large differences by continent of origin. The highest prevalence was found among European injecting drug users (54.1%; 95% CI, 37.1-70.1), South American homosexual men (41.6%; 95% CI, 32.0-51.8), and heterosexuals from Sub-Saharan Africa (8.9%; 95% CI, 7.2 10.8). CONCLUSIONS: Non-Italian patients with STIs seem to consist mainly of migrants, and STI transmission patterns differ from those among Italians; this information is important for developing targeted STI prevention efforts. PMID- 14743071 TI - Cost-effectiveness analysis of screening adolescent males for Chlamydia on admission to detention. AB - BACKGROUND: Chlamydia trachomatis infections can lead to serious and costly sequelae. Because chlamydia is most often asymptomatic, many infected youth do not seek testing. Entry to a detention system provides an opportunity to screen and treat many at-risk youth. GOAL The goal of this study was to determine the cost-effectiveness of screening male youth for chlamydia on entry to detention. STUDY DESIGN: Incremental cost-effectiveness of 3 chlamydia screening strategies was compared for a hypothetical cohort of 4000 male youth per year: 1) universal chlamydia screening using a urine-based nucleic acid amplification test (NAAT), 2) selective NAAT screening of urine leukocyte esterase (LE)-positive urines, and 3) no screening. The model incorporated programmatic costs of screening and treatment and medical cost savings from sequelae prevented in infected males and female partners. The analysis was conducted from the healthcare system perspective. RESULTS: Chlamydia prevalence in the sampled population of 594 was 4.8%, and the average number of female sexual partners/infected male was 1.6. Universal NAAT screening was the most cost-effective strategy, preventing 37 more cases of pelvic inflammatory disease (PID) and 3 more cases of epididymitis than selective screening and saving an additional 24,000 dollars. The analysis was sensitive to NAAT cost, LE sensitivity, rate of PID development, PID sequelae cost, and number of female partners. Universal screening remained the most cost saving for prevalence as low as 2.8% or higher. CONCLUSIONS: Universal chlamydia screening of adolescent males on entry to detention was the most cost-effective strategy. Savings are primarily the result of the prevention of PID in recent and future partners of index males. Screening detained male youth using a urine-based NAAT provides a public health opportunity to significantly reduce chlamydia infections in youth at risk for sexually transmitted diseases. PMID- 14743072 TI - High prevalence of anal squamous intraepithelial lesions in HIV-positive men despite the use of highly active antiretroviral therapy. AB - BACKGROUND: The impact of highly active antiretroviral therapy (HAART) on the natural history of HPV infection and anal squamous intraepithelial lesions (SIL) in HIV-infected men who have sex with men (MSM) is poorly documented. GOAL The goal of this study was to evaluate the prevalence of anal HPV infection and SIL inpatients under HAART. STUDY DESIGN: Forty-five HIV-infected protease inhibitor experienced MSM were enrolled in a cross-sectional study. Each patient provided anal samples for anal cytology, histology, and human papillomavirus (HPV) DNA testing. RESULTS: The patients had previously received HAART for a median of 32 months. Anal cytology was abnormal in 32 of 45 (71%) patients, including high grade SIL in 10 patients (22%), low-grade SIL in 19 patients (42%), and atypical squamous cells of undetermined significance in 3 patients (7%). HPV DNA was detected 36/45 men (80%). The prevalence of anal SIL and HPV infection were similar in patients exhibiting a significant increase in CD4+ cell count after HAART initiation compared with those who did not. CONCLUSION: Our results demonstrate a high prevalence of anal SIL, including high-grade SIL, and anal HPV infection in HIV-infected MSM despite immune restoration under HAART. PMID- 14743073 TI - Changes in the transmission dynamics of the HIV epidemic after the wide-scale use of antiretroviral therapy could explain increases in sexually transmitted infections: results from mathematical models. AB - BACKGROUND: Recent increases in bacterial sexually transmitted infections (STI) and risk behavior have coincided with the introduction of antiretroviral therapy (ART) in homosexual communities of industrialized countries. The reasons for these increases are not fully understood. GOAL: The goal of this study was to understand the various effects of ART on risk behaviors and STI. OBJECTIVE: The objective of this study was to assess the independent impact of the change in the transmission dynamics of HIV/AIDS as a result of the wide-scale use of ART on a bacterial STI. STUDY DESIGN: We developed a mathematical model of bacterial STI and treated/untreated HIV/AIDS infection for an open homosexual population. At the individual level, we assume that susceptible and healthy HIV-positive individuals do not increase their risk behavior as a result of ART over time. However, individuals with AIDS, who are successfully treated with ART, can resume sexual activity. The impact of the wide-scale use of ART on risky behavior, STI, and HIV/AIDS was evaluated over a wide range of assumptions on treatment use, ART efficacy, and population characteristics. RESULTS: Over 10 years, 0% to 55% new bacterial STI could be attributed to the wide-scale use of ART as a result of more modest increases (0-25%) in risky sex occurring at the population level rather than at the individual level. These increases have a negative impact on HIV if coverage is too low. Increasing treatment coverage helps to prevent more HIV infections despite larger increases in risky sex and STI that is predicted to ensue. CONCLUSION: Taking the differential impact of wide-scale use of ART into account helps to interpret recent behavioral and STI trends. Our results have implications for prevention strategies and for the formulation of public health policies. A better understanding of the differential impact of ART on sexual network over time is required. PMID- 14743074 TI - Rescreening for gonorrhea and chlamydial infection through the mail: a randomized trial. AB - BACKGROUND: Rescreening patients after treatment of Chlamydia trachomatis or Neisseria gonorrhoeae infection has had high yield but low rates of participation. GOAL: The goal of this study was to determine if rescreening for gonorrhea and chlamydial infection in a largely urban sexually transmitted disease population would be more successful if individuals were given the option of submitting a specimen for testing through the mail. STUDY DESIGN: We conducted a randomized clinical trial involving 122 patients of whom 62 were assigned to clinic rescreening and 60 were given the option of either mailing a specimen for testing or going to a clinic for rescreening. RESULTS: Twenty-seven patients (45%) given the option of either rescreening in the clinic or through the mail and 20 (32%) assigned to clinic rescreening were rescreened within 28 days of enrollment in the study (odds ratio, 1.7; 95% confidence interval, 0.8-3.8). Of the 60 patients randomized to the clinic rescreening or mailing option, 11 of 18 (61%) who opted to mail in a specimen and 16 of 42 (38%) who chose clinic rescreening were rescreened within 28 days of enrollment (P = 0.10). CONCLUSIONS: Although not statistically significant, this study indicates that mailed rescreening could be a successful method to increase rescreening rates. PMID- 14743075 TI - Perceived social cohesion and prevalence of sexually transmitted diseases. AB - BACKGROUND: Although physical attributes have been shown to be associated with sexually transmitted disease (SD) rates, there is little information about the association between social attributes and STD rates. GOAL: The objective of this study was to determine the association between gonorrhea prevalence and perceptions of social cohesion in impoverished, urban neighborhoods. STUDY DESIGN: We conducted a street-based survey of 18- to 24-year-olds residing in selected census block groups in Baltimore City, Maryland. Census block groups eligible for selection were defined as impoverished (greater than 20% in poverty) and unstable (lowest 25th percentile for stability). From the eligible census block groups, 5 from high gonorrhea rate (greater than the 75th percentile) census block groups and 5 from the lower gonorrhea rate (lowest 25th percentile to equal or greater than the 75th percentile) census block groups were randomly selected. Participants within the 10 selected census block groups were recruited using a street-intercept method. Participants were asked about perceived social cohesion and control. RESULTS: Results showed that for young adults 18 to 24 years of age residing in high gonorrhea census block groups, the mean social cohesion index scores were 1.7 points lower than mean social cohesion index scores of the participants residing in the low gonorrhea census block groups (P <0.01). CONCLUSION: Future research needs to be conducted to determine the temporal association between gonorrhea prevalence and local social cohesion dynamics. PMID- 14743076 TI - Evaluation of a Treponema pallidum-specific IgM enzyme immunoassay and Treponema pallidum western blot antibody detection in the diagnosis of maternal and congenital syphilis. AB - BACKGROUND: Congenital syphilis (CS) is a result of untreated or inadequately treated maternal syphilis. CS is more likely with early stages of maternal syphilis, but most mothers lack signs or symptoms and the risk of CS is unclear. GOAL: The goal of this study was to evaluate Treponema pallidum IgM Western blot (TP IgM WB) and a T. pallidum IgM enzyme immunoassay (TP IgM ELISA) in mothers with syphilis to determine if positive tests better indicate a risk of CS than a rapid plasma reagin titer >/=1:16. STUDY DESIGN: Ninety-seven mother-baby pairs with reactive syphilis serology were evaluated. RESULTS: TP IgM WB tests were positive in 18 pregnancies (7 of 18 babies had CS) and negative in 79 pregnancies (7 of 82 babies had CS). Thirty-two mothers had titers >/=1:16 (6 babies with CS) and 65 mothers had titers /=1:16. PMID- 14743077 TI - Congenital syphilis in Russia: the value of counting epidemiologic cases and clinical cases. AB - BACKGROUND: Congenital syphilis (CS) reports in Russia increased 26-fold from 1991 to 1999. Case reports included only infants who were clinical cases, had persistent serologic changes, or confirmed syphilitic stillbirth. Although not reported, policies stipulate that infants of inadequately treated or untreated mothers receive preventive penicillin treatment. GOAL: We examined whether risk factors and consequences for epidemiologic cases of CS (infants of inadequately treated mothers) resembled those of clinical cases and differed from those of noncases (infants of adequately treated mothers). STUDY DESIGN: A retrospective record review from Maternity Houses in 5 sites identified 715 syphilis-infected women who gave birth. RESULTS: Among women with maternal syphilis, 11% (n = 81) of infants were clinical cases, 56% (n = 402) were epidemiologic cases, and 33% (n = 232) were noncases of CS. Compared with noncases, maternal risk factors for epidemiologic cases included nonresidence (P <0.01), late syphilis (P <0.01), unemployment (P <0.01), no prenatal care (P <0.01), and syphilis testing at >/=28 weeks (P <0.01). Each of these was also significant for being a clinical case. Associated consequences of CS for the epidemiologic cases included increases in stillbirth (P <0.01), preterm birth (P <0.01), low birth weight (P <0.01), transfer to a pediatric hospital (P <0.01), and abandonment (P <0.05). Each of these except stillbirth was significantly elevated among clinical cases. Nearly half of the epidemiologic cases had no record of any penicillin treatment for the infant. Epidemiologic cases were significantly more likely than noncases to have no clinical or laboratory follow up. CONCLUSION: In Russia, maternal risk factors and perinatal consequences for epidemiologic cases of CS resembled those of clinical cases. Expanding national reporting to include epidemiologic cases would strengthen CS prevention and monitoring. PMID- 14743081 TI - [Pneumology research within the INSERM: for how long?]. PMID- 14743078 TI - Qualitative assessment of venues for purposive sampling of hard-to-reach youth: an illustration in a Latino community. AB - BACKGROUND: Latino youth suffer disproportionately from unintended pregnancy and sexually transmitted infections (STIs), but studies of the social context of their sexual behaviors are lacking. GOAL: Our qualitative assessment of recruitment venues in a Latino neighborhood had 3 objectives: to identify venues where youth at risk of unintended pregnancy and STIs could be found; to describe different youth "crowds"; and to investigate how and where youth meet their sex partners. STUDY DESIGN: We conducted ethnographic interviews with 62 youth recruited primarily from street sites. Mapping of venues was conducted with Map Info. RESULTS: Youth crowds included the Regulars, gang-related crowds, street economy affiliated crowds, and female crowds. Maps demonstrated the dominance of the venues in the Mission by gang members. Street sites are important venues for meeting sexual partners. CONCLUSION: The qualitative assessment produced insights and hypotheses that can contribute to the planning of research, outreach, testing, and interventions with Latino youth. PMID- 14743082 TI - [Does mechanical injury of the peripheral airways play a role in the genesis of COPD in smokers?]. PMID- 14743083 TI - [Clinical practice guidelines for the management of lower respiratory tract infections: are they consistent?]. PMID- 14743084 TI - [Alveolar epithelium repair: a clinically relevant challenge for respiratory medicine]. PMID- 14743085 TI - [Tuberculosis: we need to act faster]. PMID- 14743086 TI - [Smoking dependence and symptoms of anxiety and depression among Tunisian smokers]. AB - INTRODUCTION: The aim of this study was to describe the incidence of depression and anxiety, and their relationship with smoking dependence, in patients attending a Tunisian smoking cessation clinic. METHODS: We studied 72 Tunisian smokers attending the smoking-cessation clinic at the Charle Nicolle Hospital in Tunis. Nicotine dependence was assessed by the Fagestrom Test for Nicotine Dependence. Anxiety and depression symptoms were measured using the HAD (Hospital Anxiety Depression) Scale. RESULTS: The prevalence of anxiety and depression was 22.9% (16 patients) and 20% (14 patients) respectively with four patients (7.1%) exhibiting symptoms of both. Overall, 50% of the group had emotional morbidity with high HAD scores for depression or anxiety, or both. Smokers with symptoms of anxiety and/or depression had higher physical and psychological dependence, smoked more at times of stress, had a reduced quitting rate and endured more withdrawal symptoms than those smokers without anxiety or depression. CONCLUSIONS: The data of this survey from Tunisia, a country that has just put in place a tobacco control strategy, underline the high rates of anxiety and depression that exist in patients attending a smoking cessation clinic. It confirms the association between anxio-depressive disorders and a high level of smoking dependence. PMID- 14743087 TI - [Agreement between clinical practice guidelines for management of community acquired pneumonia. A retrospective study of 101 hospitalized patients]. AB - INTRODUCTION: Management guidelines for acute community acquired pneumonia vary considerably. The objective is to estimate by a retrospective study the uniformity of the recommendations for the management of patients and the choice of initial empirical antibiotic therapy. METHODS: Eight English and French language guidelines published between 1998 and 2001 were identified by a search of the literature. They were applied retrospectively to a sample of 101 patients admitted to a university hospital in 2000 with a diagnosis of pneumonia. RESULTS: Hospital admission was advocated for between 61% and 95% and admission to intensive care for between 8% and 35% of the patients, depending on the guidelines under consideration. The actual management conformed to that advocated for between 34% and 94% of the patients (kappa=0.27 [0,19; 0,34]). Compliance of the empirical antibiotic therapy (drug, dose, mode of administration) with the recommendations varied from 0% to 68% of the patients depending on the guidelines considered (kappa=0.01 [-0,10; 0,12]). CONCLUSIONS: The heterogeneity of the guidelines is manifest by important variations in the recommendations for management and initial empirical therapy. These differences are due, in part, to a paucity of evidence based data upon which to base the guidelines. It would appear essential to harmonise the guidelines in a way that is appropriate for the country of their intended use. PMID- 14743088 TI - [Validation of an asthma knowledge questionnaire]. AB - INTRODUCTION: A patient's knowledge of his disease and its treatment is an essential part of the evaluation of an educational process. It is useful therefore to use a rapid, easy and valid means of collecting the information necessary to produce an educational programme adapted to the needs of the patient. METHODS: Following a review of the literature an initial questionnaire was constructed. It included four domains: biomedical, signs and symptoms of severity, general knowledge and treatment. The questionnaire was administered to 73 asthmatics in order to assess its clarity and comprehensibility. It was then reviewed and modified in both format and content by 10 French experts. The revised questionnaire was completed by 108 asthmatics distributed throughout 10 French respiratory centres, a group of 83 non-asthmatic subjects and 203 sixth year medical students at the Bordeaux University School of Medicine. RESULTS: The mean scores were: 19 for the non-asthmatics (range 2-36), 25.7 for the asthmatics (range 4-38) and 32.9 for the students (range 17-40), p 0.0001. The questionnaire was shown to be discriminating with good reliability and reproducibility: alpha Cronbach coefficient=0.82; intra-class correlation coefficient=0.70. CONCLUSIONS: This study has validated a French language asthma knowledge questionnaire. PMID- 14743089 TI - [An experimental study on the systemic distribution of calibrated talc after intra-pleural injection]. AB - INTRODUCTION: Among the agents used to produce pleural symphysis talc is the most effective and least expensive. However, its use is controversial on account of the description of respiratory complications associated with subsequent systemic spread of the talc particles. This hypothesis rests on clinical and experimental observations of talc particles in the viscera. However, all talc preparations are not identical and this extra-pleural spread could be dependent on particle size. This experimental study was undertaken to determine whether there was systemic spread of a calibrated talc preparation used routinely in clinical practice following intra-pleural administration in rats. METHODS: 48 rats received 20 mg (11 rats) and 40 mg (33 rats) of calibrated talc suspended in 1 ml of physiological saline by intra-pleural injection. The animals were randomised for sacrifice at 24 hours (22 rats) and 72 hours (22 rats) after the injection. The lungs, parietal pleura, diaphragm, liver, spleen, pericardium, brain and blood were examined by light microscopy and polarised light to search for bi-refringent particles. RESULTS: No deaths occurred during the procedure. At the time of sacrifice no pleural symphysis was seen. In 5 animals some talc particles were seen in the extra-thoracic organs: in the liver in 3 in the spleen in 1 and one particle in the brain of one animal examined by electron microscopy. No talc particles were found in the blood. CONCLUSIONS: Intra-pleural injection of calibrated talc, (Steritalc-Novatech-Plan de Grasse-France) has a weak systemic spread in > small animals. These results may be related to the diameter of the talc particles used (mean 33.6 microns; median 31.3 microns). The hypothesis that systemic spread is influenced by the diameter of the talc particles needs to be supported by experimental studies using talc particles of smaller diameter in order to compare the systemic distribution of the different preparations. PMID- 14743090 TI - [Bioenergetic characteristics of the diaphragm after spinal cord injury]. AB - INTRODUCTION: C2 spinal cord hemisection interrupts descending bulbospinal respiratory axons coursing through the cervical spinal cord. One consequence of this type of spinal cord injury is the partial deafferentation of the ipsilateral phrenic nucleus, which renders the hemidiaphragm paralysed. The biochemical properties of the intact or denervated locomotor skeletal muscles have been studied extensively, whereas the biochemical properties of the diaphragm after a cervical spinal cord injury received relatively little attention. This seems unfortunate, given that the diaphragm is the most important mammalian respiratory muscle and is the only skeletal muscle considered essential for survival. Therefore, the purpose of this work was to analyse the bioenergetic characteristics of the diaphragm following a cervical spinal cord injury. METHODS: Segments of the crural diaphragm from rat were homogenised to determine the activities of a glycolytic enzyme, lactate dehydrogenase (LDH), and a Krebs cycle enzyme, citrate synthase (CS). RESULTS: Data show a significant decrease of the CS activity on the ipsilateral hemidiaphragm to the cervical hemisection. No change in the LDH is observed between the animal without or with a cervical spinal cord injury. CONCLUSIONS: These data suggested that the CS activity could be dependent of the nervous influx from the central respiratory rhythm generator and that LDH activity could be maintained by a peripheral respiratory activity or by the cross phrenic phenomenon activation. Biochemical data should be considered in subjects presenting respiratory deficiency induced by a cervical spinal cord injury. Furthermore, the diaphragmatic muscle metabolic activity could be used to evaluate the functional respiratory recovery observed spontaneously or experimentally after using repair strategies of the spinal cord. PMID- 14743091 TI - [Keratinocyte growth factor and Hepatocyte growth factor: their roles in alveolar epithelial repair]. AB - INTRODUCTION: The alveolar epithelium is the principal target in the course of acute lung injury (ALI) and the acute respiratory distress syndrome (ARDS) and to a lesser extent in chronic reactions like pulmonary fibrosis. STATE OF ART: Keratinocyte growth factor (KGF) and Hepatocyte growth factor (HGF) are potent growth factors for type II pneumocytes and seem to play a specific role in the process of alveolar repair. PERSPECTIVES: The studies conducted by our group have demonstrated 1) that KGF and HGF are present in biologically active concentrations in human pulmonary alveoli in ALI and ARDS, 2) that in these patients as well as those with pulmonary fibrosis circulating neutrophils are an important source of HGF. HGF and KGF act within a system involving other factors such as parathyroid hormone related protein (PTHrP). PTHrP acts in an autocrine and paracrine manner to regulate the balance between proliferation and apoptosis of alveolar epithelial cells. CONCLUSIONS: Our results obtained in humans suggest a beneficial role for neutrophils in the alveolar repair after acute or chronic lung injury. The experimental data suggest that use of KGF and HGF might be considered in the future in the treatment of human acute or chronic lung injury. PMID- 14743092 TI - [Ventilatory response to maximal exercise in healthy children]. AB - INTRODUCTION: The evaluation of the ventilatory response of children during exercise is essential to determine its role in impaired exercise tolerance. The aim of this review is to describe the variables and the values of maximal ventilatory parameters observed in healthy children in the published literature. STATE OF ART: The maximal ventilation (VEmax) and the tidal volume (VTmax) increase in a linear fashion with age and plateau in boys at 15 years, and in girls at 13 years. The main variables for the parameters connected to volume- VEmax and VTmax--are anthropometric characteristics, in particular, the lean body mass. Most studies show a value of 30 ml.kg(-1) for a VTmax on the total body mass in pre-puberty and a slight increase thereafter. The ventilatory reserves and the VTmax on vital capacity increase with age until respective values of 30% and 50% are reached at 17 years. The maximal parameters connected to time are independent of anthropometric characteristics. The TI/TTOT ratio (inspiratory time to total time of the respiratory cycle) is stable with a value of 0.5. The maximal respiratory frequency decreases slightly with age without differences between the genders. PERSPECTIVES AND CONCLUSION: Only studies of larger numbers of children, proposing relationships derived from allometric equations, will be able to provide real reference values. PMID- 14743093 TI - [Tuberculosis in elderly persons living in institutions]. AB - INTRODUCTION: In industrialised countries the incidence of tuberculosis increases with age. STATE OF ART: Tuberculosis remains a current problem in the elderly and may be contracted in an institution. Elderly persons living in institutions account for four times more cases than the elderly living at home. Tuberculosis may present in all forms and situations but the clinical and radiological features may be atypical. Although tuberculin anergy may be found frequently in elderly patients the diagnosis relies on the same principles as in the younger patient. Mortality, even with treatment, is increased and rises with age. Several problems arise in institutions: the screening of sick residents and of subjects at risk as well as the screening of infected patients. PERSPECTIVES: A rigorous policy of prevention and surveillance should lead to a fall in the incidence of the disease in institutions. CONCLUSIONS: This collection of data demonstrates that tuberculosis remains a real health problem in elderly subjects in institutions. PMID- 14743094 TI - [Clinical practice guidelines: a reader's guide]. AB - INTRODUCTION: This article is devoted to the critical appraisal of clinical guidelines. METHODS: The example chosen is the update of the guideline "Clinical management of chronic obstructive pulmonary disease: diagnosis, follow-up, and acute exacerbations". The full and abbreviated versions of the text published by the "Societe de Pneumologie de la Langue Francaise" were analysed by a guide for the critical evaluation of clinical guidelines. BACKGROUND: The evaluation guide "AGREE" allows assessment of six areas included in the development of a guideline: scope and objectives, involvement of the groups concerned, rigour of development, clarity and presentation, applicability and editorial independence. CONCLUSION: Clinical guidelines are going to play an important part in medical practice. Evaluation of professional practice will be based on their contents. A critical appraisal of their validity and applicability in this way allows an accurate decision on their subsequent application. PMID- 14743095 TI - [Secretory immunity of the airways]. AB - INTRODUCTION: Continuous exposure of the respiratory tract to inhaled particles and microbes implies the presence of effective defence mechanisms at a bronchial and alveolar level. STATE OF ART: Among the mechanisms involved secretory mucosal immunity contributes considerably to the defence of the bronchial tree. This immunity depends essentially on the active trans-epithelial transport of IgA involved in both innate non-specific and acquired specific immunity. Recently an IgA receptor has been identified on the surface of phagocytes including alveolar macrophages, establishing a link between alveolar and bronchial defences. PERSPECTIVES: The respiratory mucosa represents a crucial interface between the host and its environment, and should provide in the future a new target for the development of diagnostic and therapeutic tools. CONCLUSIONS: Beyond its function as an anatomical barrier the bronchial epithelium possesses a secretory activity that is essential for the protection of the lung. Despite a better understanding of mucosal immunity this secretory activity and in particular the part played by IgA remains to be elucidated. PMID- 14743096 TI - [VENISE: Non-invasive ventilation during mechanical ventilation weaning in chronic respiratory failure patients. A prospective randomised controlled and multicenter trial]. AB - INTRODUCTION: The clinical efficacy of non-invasive ventilation (NIV) has now been demonstrated in the management of acute-on-chronic respiratory failure (ACRF) in various etiologies. Endotracheal mechanical ventilation (ETMV) can lead to numerous complications and weaning difficulties increasing the risk of prolonged ETMV, morbidity and mortality as well as excess cost of intensive care. Therefore, it could be useful to consider NIV for delivering effective ventilatory support to reduce the length of ETMV in ACRF patients who are still not capable of maintaining spontaneous breathing. From the interesting but discordant results of two recent randomised controlled trials, a working group from the Societe de Reanimation de Langue Francaise (SRLF) decided to perform a new prospective randomised controlled multicenter trial. The aim of the study will be to assess the usefulness of NIV as an extubation and weaning technique in ventilated ACRF patients. METHODS: The methodology used will compare three parallel weaning strategies in ACRF patients considered difficult to wean: invasive conventional weaning (group A), extubation relayed by nasal oxygentherapy (group B), and extubation relayed by NIV (group C). Based on the main end-point defined as the weaning success rate, 208 patients from 17 investigator centers are planned to be included. Results of the study will also allow to assess the respective impact of the three weaning strategies on the length of ETMV and weaning, the mechanical ventilation-related morbidity, the patients lengths of stay and mortality. EXPECTED RESULTS: Results of the VENISE trial should permit to improve the management of the difficult to wean ACRF patients and thus contribute to more precisely define the place of NIV in the weaning and prevention of re-intubation strategies in these patients. PMID- 14743097 TI - [How to describe the distribution of a variable? Normality tests and management of extreme values]. PMID- 14743098 TI - [What is a parametric test?]. PMID- 14743099 TI - [What is a non-parametric test?]. PMID- 14743100 TI - [Pneumonitis caused by hydrocarbon inhalation]. AB - INTRODUCTION: Kerosene pneumonitis is rare. It most commonly occurs in children who have ingested a hydrocarbon by accident. In adults, pneumonitis caused by aspiration of incompletely vaporized kerosene may be seen in fire-eaters can cause pneumonitis. CASE REPORTS: We report six cases of fire-eater's pneumonitis, from our region, due to hydrocarbon inhalation. CONCLUSIONS: This inhalational pulmonary insult has classical clinical manifestations. The frequency of cases, particularly in tourist areas that host street festivals, is probably underestimated. Its tendency to spontaneously resolve questions the need for antibiotics and corticosteroids. PMID- 14743101 TI - [Spontaneous pneumomediastinum in dermatomyositis: a pathophysiological approach using imaging]. AB - INTRODUCTION: Spontaneous pneumomediastinum is a rare complication of dermatomyositis. CASE REPORT: We report a case of pneumomediastinum with massive subcutaneous emphysema occurring in a female patient with dermatomyositis treated with cortico-steroids. CONCLUSIONS: Our case illustrates perfectly the mechanism of spread of air along the broncho-vascular structures and also explains the presence of pneumomediastinum in the absence of pneumothorax. PMID- 14743102 TI - [A favourable outcome in yellow nail syndrome: role of respiratory physiotherapy]. AB - INTRODUCTION: The yellow nail syndrome is a rare condition that is easily diagnosed but the nail manifestations are poorly understood. CASE REPORT: A 51 year old patient presented with a chronic cough. The diagnosis was based on the typical appearance of the nails. The patient had bilateral basal bronchiectasis. Daily physiotherapy with bronchial drainage lead to a progressive improvement in the respiratory symptoms without recourse to antibiotics. Surprisingly the abnormalities of the nails disappeared after 2 years treatment. CONCLUSION: This observation illustrates the possibility of spontaneous resolution of severe nail abnormalities during the course of the yellow nail syndrome. PMID- 14743103 TI - [Needle aspiration in paradoxical hypertrophy of tuberculous lymphadenitis]. AB - INTRODUCTION: Paradoxical hypertrophy is frequent in case of peripheral lymph node tuberculosis. Its management remains controversial. CASE STUDY: We report a case of susclavicular and anterior cervical lymph nodes tuberculosis complicated by paradoxical hypertrophy after 4 months of treatment in a 59-year-old woman. The lymph node was not compressive but was limiting the cervical mobilisation. The caseum was sucking three times with a fine needle. Thereafter we performed an isoniazid injection each 24 hours for three days at three different lymph node sites. Antituberculous treatment was reinitiated for 10 months at all and lymph nodes completely disappeared without functional sequels. CONCLUSION: Paradoxical hypertrophy is a frequent complication of peripheral lymph node tuberculosis. Caseum suction of the paradoxical hypertrophy with a fine needle may constitute an alternative treatment of surgery. PMID- 14743104 TI - [Respiratory sinus arrhythmia and voluntary control of breathing]. PMID- 14743105 TI - [End-of-life care in the intensive care unit. Recommendations of the Societe de Reanimation de Langue Francaise]. PMID- 14743107 TI - [An iconoclast's view of onychomycosis and their treatment]. PMID- 14743108 TI - [Can we evaluate the frequency of onychomycosis?]. AB - Onychomycosis represents one of the most common nail diseases. Difficult to bear by some patients, onychomycosis is one of the most frequent reasons for consulting by lack of uniformity and group-control a mycological centre. However, frequency of onychomycosis is miss appreciable. The studies of prevalence concern the general population (2 to 13 per cent according to the different authors) consulting a medical practitioner or specialist (dermatologist). Recent surveys done a large scale in Europe in adults indicated high prevalence: 20 to 30 per cent, depending on whether the investigator is a general practitioner or a dermatologist. In any case, all the studies concur in the opinion that onychomycosis has been in constant progression over the last twenty years. Rarely observed in children, frequent in adults, onychomycosis principally affects particularly the elderly. In Western Europe and in North America, onychomycosis involves principally the feet--especially in males. By contrast, in Southern Europe, in the Middle and Far East, the prevalence is highest in women's fingernails (often associated with paronychia). Among the factors promoting fungal nail invasion, some are local depending on the patient (trophic troubles and circulatory impairment overlapping of digits, etc.), some are due to general factors such as immunosuppression, diabetes mellitus or psoriasis. There are also behavioural factors (occupation, lifestyle, sports) which favor the meeting with the pathogenic fungus. PMID- 14743109 TI - [Normal and mycotic nail]. AB - Knowledge of anatomy and physiology of the nail unit is necessary to understand onychomycosis, their clinical patterns, and response to the different treatments. It was useful for the discovery of new transungueal products. In this chapter, we describe anatomy and physiology of the normal nail and the main routes of fungus entry that lead to the onychomycosis. PMID- 14743110 TI - [Clinical types of onychomycosis]. AB - Clinical classification of onychomycosis is based on how the pathogenic agent penetrates the unguis. The disto-lateral sub-ungual variety is the most common. Dermatophytes (especially Trichophyton rubrum) and Scytalidium in tropical regions (Sc hyalium, Sc dimidiatum) are the most frequent toenail pathogens. Progression of a yellow friable sub-ungual hyperkeratosis associated with longitudinal striations and/or xanthonychial or leuconychial zones (sometimes pigmented with Trichophyton rubrum nigricans or Scopulariopsis brevicaulis) lead rapidly to onycholysis and later to total dystrophy of the ungual plate. Associated paronychia is more common in scytalidiasis. Candida species (tropicalis, parapsilosis) and fungi (Aspergillus sp. Fusarium sp, Acremonium sp, Penicillium sp, Scopulariopsis brevicaulis) generally colonize pre-existing onycholysis. For the fingernails, candidal colonizations secondary to pre existing onycholysis is much more frequent than primary dermatophyte or scytalidium onychomycoses which are much less hyperkeratotic than on toenails. The one hand two feet tinea syndrome caused by Trichophyton rubrum is a particular entity. Proximal sub-ungual onychomycoses without fingernail or toenail paronychia is generally caused by Trichophyton rubrum in immunodepressed subjects (AIDS). Initial proximal leuconychia progresses to the distal part of the nail. Proximal lesions associating proximal leuconychia and paronychia result from fungi, Fusarium being the most commonly identified agent. Onyxis complicating chronic paronychia, generally related to Candida colonization, occurs in subjects with particular conditions (immunodepression, distal vascular disorders). Superficial onychomycosis, e.g. superficial toenail leuconychia, is mainly due to Trichophyton interdigitale, more exceptionally to Trichophyton rubrum (children, immunodepressed), and rarely Candida (children). Endonyx onychomycosis occurs when the pathogen invades the entire thickness of the nail (milky leuconychia without sub-ungual hyperkeratosis). Trichopnyton violaceum or soudanense is the most common pathogen. Even though the clinical presentation of onychomycosis is highly suggestive of the pathogenic agent, the lack of specific criteria implies a mycological sample to confirm the diagnosis and identify the causal agent before initiating treatment. PMID- 14743111 TI - [Recommended techniques for obtaining nail specimens and mycologic diagnosis of onychomycosis]. AB - The diagnosis of onychomycosis requires an assessment of both clinical and laboratory features. The laboratory diagnosis is essential to confirm the fungal origin of the onyxis. Various types of onychomycosis need different methods to obtain a nail specimen. It requires a direct examination to confirm the presence of fungi, and a fungal culture to identify the specific genus and species of the pathogen. A good interpretation of these results is necessary to adapt the therapeutics. PMID- 14743112 TI - [Differential diagnosis of onychomycosis]. AB - In order to give an effective treatment to a patient with nail disorders, a correct aetiological diagnosis is necessary. Distal and lateral thickening of nail bed and nail plate with onycholysis, deep or superficial leuconychia, melanonychia, paronychia, distal and lateral onycholysis, and total dystrophic onychomycosis are the common physical signs of onychomycosis which represent 50 per cent out of nails disorders. Nail disorders due to cutaneous or general diseases with similar alteration of nail configuration may be confused with onychomycosis. In many cases, an aetiological trend is given by a total body examination of the skin and the mucosa. To diagnose correctly a non fungal nail disorder from an onychomycosis sometimes is a real challenge. According non fungal nail disorder and fungal nail infection may be associated, so a mycological examination and sometimes histopathological examination are very helpful to establish a good diagnosis. PMID- 14743113 TI - [Nail histomycology. Protean aspects of a human fungal bed]. AB - Onychomycosis exhibits considerable diversity when the disease is scrutinized using traditional, fluorescence or confocal microscopy. Histomycology is a non invasive assessment performed on nail clippings. The location of and the density in fungal cells is variable. In some instances, these aspects remain clinically unsuspected. In vivo confocal microscopy can provide the same information. Computerized image analysis of histological sections is the most powerful means for quantifying the fungal load. Immunohistochemistry provides information about the identity of the fungus or the association of different fungi present in the nail plate. Mixed infections may be unifocal or located at different levels in the nail apparatus. The viability of fungi as assessed by vital stains can be visualized under the microscope and quantified by flow cytometry. The different aspects of nail histomycology are complementary and shed some light on sometimes unsuspected aspects of onychomycoses. PMID- 14743114 TI - [The management of onychomycosis]. AB - Despite the appearance of the new systemic drugs there is still an incompressible failure rate in treating patients affected by onychomycosis. Moreover some patients are unable or unwilling to take oral drugs. They perceive the infection to be too trivial to deserve systemic therapy. In addition the use of the new potent systemic drugs may sometimes be limited by adverse effects and drug-drug interactions. A rationale for a stepped therapeutic approach to the treatment of onychomycosis is suggested using the transungual antifungal delivery systems, the systemic antifungal drugs and surgical or chemical debridement. Long-term management with the nail lacquers applied twice a month should still improve the cure rate and its economic implication. PMID- 14743115 TI - [Pharmacoeconomic aspects of the treatment of onychomycosis]. AB - The clinical management of severe onychomycosis is known to be a long and difficult task. When deciding which treatment to prescribe to our patients we may be tempted to use the efficacy rate as the only objective measurement or on the other hand to assess treatment costs only by looking at the drug acquisition cost. The role of pharmacoeconomics is to provide a method that evaluates outcomes and costs at the same time; it is an aid to better decision making. Cost effectiveness studies take into account the efficacy of each treatment regimen balanced against the global cost of treatment. So we can assess the number of patients cured for a given health care budget. Two recent studies presenting the results of a combination therapy of an antifungal therapy and antifungal lacquer give us the opportunity to illustrate some pharmacoeconomic principals. PMID- 14743116 TI - [Onychomycosis and quality of life]. AB - The study of the quality of life in onychomycosis is recent. Since 10 years, the evolution of methods has allowed to have more precision about the patient's feelings, especially in physical, functional, psychical and social criteria. In the future, we can hope than such studies could help to manage therapies decisions to get a best efficacy and lower costs. PMID- 14743118 TI - Use of severity score to evaluate quality of care. PMID- 14743119 TI - Combined general and epidural anesthesia with ropivacaine for renal transplantation. AB - AIM: To evaluate the effectiveness and safety of epidural ropivacaine anesthesia in association with light general anesthesia during renal transplantation and compare epidural and endovenous analgesia techniques for postoperative pain control. METHODS: EXPERIMENTAL DESIGN: prospective randomized study. SETTING: Organ Transplantation Center, Department of Surgery, "Tor Vergata" University of Rome, St. Eugenio Hospital, Rome. PATIENTS: 25 patients affected by chronic renal failure were enrolled in this study. Thirteen constituted the combined epidural general anesthesia group (EPI-GEN), mean age 40.15+/-9.81 years; while the others constituted the general anesthesia group (GEN), mean age 46.75+/-7.45 years. Operation: cadaveric renal transplantation. Group EPI-GEN: epidural anesthesia performed with 12-15 ml of a ropivacaine 0.75% and fentanyl 5 microg/ml solution followed by light intravenous or inhalatory general anesthesia and postoperative epidural analgesia with ropivacaine 0.2% and fentanyl 2 mg/ml. Group GEN: inhalatory or intravenous general anesthesia and intravenous tramadol postoperative analgesia. MEASUREMENTS: hemo-dynamics, renal function, arterial blood gases analysis, acid-base balance and postoperative pain data was collected and examined. RESULTS: Postoperative epidural analgesia resulted significantly more effective than intravenous tramadol. PaO(2)/FiO(2) ratio was significantly higher in group EPI-GEN patients both on awakening and throughout postoperative observation. Hemodynamics and renal function did not appear to differ significantly. CONCLUSION: Combined epidural-general anesthesia is as valid a technique as any for renal transplantation; however postoperative epidural ropivacaine analgesia resulted more effective than intravenous tramadol. Respiratory function appeared less affected, facilitating a fast and uncomplicated postoperative recovery. PMID- 14743120 TI - Intraoperative effects of combined versus general anesthesia during major liver surgery. AB - AIM: The study compares the intraoperative effects of combined versus general anesthesia during major liver surgery. METHODS: In this prospective randomized study, 70 patients were divided into 2 group of 35 subjects. Group A received general anesthesia (thiopentone, fentanyl, vecuronium, sevoflurane in a closed circuit) 15 minutes after placement of an epidural catheter (D9-D10) and induction of epidural anesthesia (6 ml 2% naropine). Continuous epidural infusion was initiated before surgical incision and continued with 0.2% naropine (7 ml/h) until the end of the operation. Group B received combined intraoperative anesthesia wit fentanyl doses according to hemodynamic parameters and 0.1 mg/kg morphine 30-4 minutes before cutaneous suture. Hemodynamic values were measured at base line (T0), and then at 15, 30, 60, 120 and 180 minutes after induction of general anesthesia (T1, T2, T3, T4 and T5, respectively). On recovery, patients were assessed for pain at rest and on movement reported on a visual analog scale; degree of motor blockade according to the Bromage scale; appearance of side effects; use af analgesic. RESULTS: A statistically significant decrease in the mean arterial blood pressure (ABP) and heart rate (HR) was noted within each group at 15 minutes after induction of general anesthesia. Significant differences in ABP were found between the 2 groups at T1 to T5, whereas HR values were substantially similar. The mean intraoperative use of fentanyl was significantly higher in Group B than in Group A, as was that of vecuronium. Pain intensity on recovery in patients who received epidural anesthesia was lower both at rest and on movement; only the patients in Group B required additional analgesics. No motor blockade was observed in either group. Nausea and vomiting were more frequent in Group B; hypotension was more frequent in Group A. CONCLUSION: The study confirms the safety of locoregional anesthesia in liver surgery, with good hemodynamic stability and absence of major side effects. The lower intraoperative use of opioids and muscle relaxants in patients who received epidural anesthesia confirms the neurovegetative protection this method provides. The data support the hypothesis that greater intraoperative use of opioids may be responsible for the higher incidence of side effects. Therefore, the intraoperative use of combined low-concentration anesthetic agents alone appears to offer a reasonable treatment option that provides adequate pain control at recovery from general anesthesia, with only minor side effects typically associated with analgesic (motor blockade) and opioids (nausea and vomiting). Given the complications associated with the technique, it should be performed by an expert anesthetist. PMID- 14743121 TI - To verify four 5-year-old mathematical models to predict the outcome of ICU patients. AB - AIM: The aim of this study is to verify calibration and discrimination after 5 years in the case mix of patients admitted to the Intensive Care Unit (ICU) during the year 2000. In this way we want to perform a quality control of our ICU in order to justify the increased amount of money spent for intensive care. METHODS: A prospective study has been made on the 357 patients admitted to the ICU during the year 2000. The Apache II score was calculated within the first 24 hours and, depending on the length of stay in the ICU, on the 5(th), 10(th) and 15(th) day after ICU admission. On the basis of the 4 mathematical models death risk has been calculated for each of the 4 times. The Hosmer-Lemeshow test was performed for calibration and ROC curves for discrimination, always for each of the 4 mathematical models. RESULTS: The 1(st) model, at 24 hours from ICU admission, showed a bad calibration (p=0.000088), while the ROC curve was 0.744+/ 0.32. Also the 2(nd) model, at the 5(th) day from admission, showed a bad calibration (p=0.000588), with ROC curve of 0.827+/-0.04. The 3(rd) model (10(th) day), was well calibrated (p=0.112247) and discriminating (ROC=0.888 +/-0.04). Finally the models at 15 days showed again a bad calibration (p=0.001422) but a very good discrimination (area=0.906+/-0.06). CONCLUSION: Developing mathematical models to predict mortality within ICUs can be useful to assess quality of care, even if these models should not be the only ICU quality controls, but must be accompanied by other indicators, looking at quality of life of the patients after ICU discharge. PMID- 14743122 TI - Monitoring of intrathoracic volemia and cardiac output in critically ill children. AB - AIM: Hemodynamic monitoring is an important step in the management of critically ill children despite the difficulty in measuring preload indices continuously. The aim of the study was to analyze cardiac output parameters and preload indices after acute changes in mean airway pressure and volemia. METHODS: Twenty-three children treated at our unit were enrolled in a prospective non randomized cohort study. Respiration was supported by controlled mechanical ventilation with positive expiratory-end pressure (PEEP), peak inspiratory pressure <20 cm H(2)O and mean airway pressure <10 cm H(2)O, and hemodynamic monitoring using the PiCCO system. Hemodynamic parameters were measured at T0 (base line), T(1) (after an increase in PEEP of 5 cm H(2)O for 10 min), and T(2) (after fluid challenge). The statistical analysis (BMPD New System software package) comprised comparison of changes at T(0) vs T(1), T(1) vs T(2) and T(0) vs T(2), construction of 3 correlation matrices and multiple linear regression analysis. RESULTS: Sixty-nine hemodynamic parameters were measured in the 23 patients. A comparison between T(0) and T(1) showed no significant changes; differences between T(0) and T(2) were found for cardiac index (CI), (p=0.003); between T(0) and T(2) significant differences were found for CI (p=0.0015), intrathoracic blood volume index (ITBVI) (p=0.04) and stroke volume index (SVI) (p=0.06). The analysis of the correlation matrices yielded ITBVI with CI (p=0.0006), ITBVI with SVI (p=1 x 10( 5)), CI with SVI (p=0.002); a significant correlation between CI and extravascular lung water index (EVLWI) was found only at T(1). Multiple linear regression analysis showed that ITBVI and SVI were predictive for variance of CI at each time point. CONCLUSION: ITBVI measured by a volumetric monitoring system such as the PiCCO may be considered a sensitive preload indicator also in critically ill children. PMID- 14743123 TI - Post-traumatic superior sagittal sinus thrombosis. Case report and analysis of the international literature. AB - The objective of this study is to focus attention on cerebral venous sinus thrombosis (CVST), a rather infrequent disease, especially when following closed head injury. Consequently we started from the clinical case report, concerning a patient admitted to our polyvalent ICU in the Hospital of Avezzano (AQ), Italy. The patient was a 15-year-old girl, that developed superior sagittal sinus (SSS) thrombosis following closed head injury (pedestrian run down by a car): owing to slow and progressive onset of deep coma with severe intracranial hypertension, emergency decompressive craniectomy was performed. The result was satisfying: patient conditions progressively improved, with return to consciousness, to good mobility and to good mental status. At present, 1 year after trauma, only mild disability is left over (right hand prehensile strength loss, and slightly moving gait). In conclusion, considering the literature data (intracerebral haematoma and deep coma are poor outcome predictors) and clinical evolution, we decided an aggressive surgical approach to save the patient's life, with satisfying results. PMID- 14743124 TI - Secondary migration of a central venous catheter. A case report. AB - A case of central venous catheter (CVC) secondary migration in a patient with Hodgkin's lymphoma is reported. The catheter was inserted in the right internal jugular vein with anterior approach. The correct position of the catheter tip in the superior vena cava was confirmed by X-ray. Secondary migration to the right subclavian vein, without displacement at the point of insertion, was reported 8 days later by a chest X-ray performed for worsening of the respiratory condition. CVC was removed and reinserted with the same procedure. The correct position of the catheter tip was confirmed by thoracic radiography till 10 days later. Epidemiological data present in the literature and secondary migration predisposing factors are reported. PMID- 14743125 TI - Presidential address: the next quarter. PMID- 14743126 TI - Endovascular management of juxtarenal aneurysms with fenestrated endovascular grafting. AB - PURPOSE: To evaluate the technical feasibility and short-term results of juxtarenal aneurysm repair with an endovascular graft that incorporated the visceral aortic segment with graft material. METHODS: Patients were studied prospectively after the implantation of an endovascular device with graft material extending proximal to the renal arteries, variably incorporating the superior mesenteric and celiac arteries. All patients were deemed to be high risk with respect to open surgical repair and had compromised proximal neck anatomy. Proximal neck lengths were or=3; 63% vs 48% and 54%; P =.0002-P <.0001). Device-related (type I) endoleaks were more frequently observed at early postoperative arteriography in group C compared with groups A and B (9.9% vs 3.7% and 6.8%; P =.01-P <.0001). Postoperatively systemic complications were more frequently present in group C (17.4% vs 12.0% in group A and 12.6% in group B; P <.0001 and.001). The first-month mortality was approximately twice as high in group C compared with the other groups combined (4.1% vs 2.1%; P <.0001). Late rupture was most frequent in group C. Follow-up results at midterm were less favorable in groups C and B compared with group A (freedom from rupture, 90%, 98%, and 98% at 4 years in groups C, B, and A, respectively; P <.0001 for group C vs groups A and B). Aneurysm-related death was highest in group C (88% freedom at 4 years, compared with 95% in group B and 97% in A; P =.001 and P <.0001, respectively; group B vs A, P =.004). The annual rate of aneurysm-related death in group C was 1% in the first 3 years, but accelerated to 8.0% in the fourth year. Incidence of unrelated death also was higher in groups C and B than in group A (76% and 82% freedom at 4 years vs 87%; P <.0001 for both comparisons). Ratio of aneurysm-related to unrelated death was 23%, 21%, and 50% in groups A, B, and C, respectively. Cox models demonstrated that the correlation between large aneurysms (group C) and all assessed outcome events was independent and highly significant. Older generation devices had an independent association with aneurysm-related and unrelated deaths (P =.02 and P =.04, respectively). However, this correlation was less strong than large aneurysm diameter (P =.0001 and P =.0009, respectively). CONCLUSIONS: The midterm outcome of large aneurysms after EVAR was associated with increased rates of aneurysm-related death, unrelated death, and rupture. Reports of EVAR should stratify their outcomes according to the diameter of the aneurysm. Large aneurysms need a more rigorous post-EVAR surveillance schedule than do smaller aneurysms. In small aneurysms EVAR was associated with excellent outcome. This finding may justify reappraisal of currently accepted management strategies. PMID- 14743128 TI - Outcomes of secondary interventions after abdominal aortic aneurysm endovascular repair. AB - PURPOSE: We assessed the distribution of secondary interventions after aortic stent grafting (EVAR) performed to treat infrarenal abdominal aortic aneurysm (AAA), and evaluated clinical success and survival in patients who underwent a secondary procedure (group 2) compared with patients who did not undergo a secondary procedure (group 1). METHODS: Two hundred fifty patients (mean age, 71.3 years) with asymptomatic AAAs (mean aneurysm diameter, 54.5 mm) underwent treatment with commercially available stent grafts. Mean follow-up was 28 months (median, 25 months). Secondary procedures were defined as any additional procedures performed after initial graft placement to treat endoleak, migration, kinking, stenosis, or occlusion. Overall clinical success was defined according to reporting standards of the Society for Vascular Surgery/American Association for Vascular Surgery. RESULTS: Sixty-eight patients (27%) required 112 secondary procedures, with a mean time from initial graft placement of 18.2 months. Patients who received grafts since removed from the market required more secondary procedures (59%, procedure:patient ratio) compared with patients who received devices still on the market (21%; P =.001). Thirty-six patients (53%) required a single secondary procedure, 24 patients (35%) required two procedures, 5 patients (10%) required three procedures, 2 patients (3%) required four procedures, and 1 patient required five secondary procedures. Ninety-eight procedures (87%) were performed with endovascular methods, including placement of 42 additional covered stent grafts (36 iliac, 6 aortic), with a success rate of 85%; 35 embolization procedures (21 lumbar, 9 internal iliac artery, 5 mesenteric), with only 23 (65%) successful; 14 angioplasty procedures, with 85% successful; 4 thrombolysis procedures, 2 of them successful (50%); and 3 successfully placed new endografts within a previous endovascular graft. Surgical secondary operations included nine femorofemoral bypass procedures and three femoral thromboendarterectomies, all of which remain patent; one cerclage of an external iliac limb; and one laparoscopic repair of a type II endoleak, which was successful. Overall clinical success rate for EVAR was 84% (211 of 250) in this series. Clinical success rate in groups 1 and 2 was 91% (166 of 182) versus 66% (45 of 68; P =.001) if all endoleaks on the most recent computed tomography scans are taken into account, and 94% (171 of 182) versus 76% (52 of 68; P =.001) if type II endoleak without aneurysm growth is not considered failure. The survival rate and rupture-free survival in groups 1 and 2 were, respectively, 97.7% +/- 1.0% and 98.5% +/- 1.4% at 1 month, 95.9% +/- 1.5% and 96.9% +/- 2.1% at 6 months, 94.4% +/- 2.0% and 93.2% +/- 3.4% at 1 year, and 80.8% +/- 5.2% and 88.5% +/- 5.0% at 3 years (P =.273, log-rank test). CONCLUSION: With close follow-up and a significant number of secondary operations, this 8-year experience has not included any aneurysm ruptures to date. Secondary operations did not lead to increased mortality, but were associated with more surgical conversions and with a higher clinical failure rate. PMID- 14743129 TI - Type II endoleak after endovascular abdominal aortic aneurysm repair: a conservative approach with selective intervention is safe and cost-effective. AB - OBJECTIVES: The conservative versus therapeutic approach to type II endoleak after endovascular repair of abdominal aortic aneurysm (EVAR) has been controversial. The purpose of this study was to evaluate the safety and cost effectiveness of the conservative approach of embolizing type II endoleak only when persistent for more than 6 months and associated with aneurysm sac growth of 5 mm or more. METHODS: Data for 486 consecutive patients who underwent EVAR were analyzed for incidence and outcome of type II endoleaks. Spiral computed tomography (CT) scans were reviewed, and patient outcome was evaluated at either office visit or telephone contact. Patients with new or late-appearing type II endoleak were evaluated with spiral CT at 6-month intervals to evaluate both persistence of the endoleak and size of the aneurysm sac. Persistent (>or=6 months) type II endoleak and aneurysm sac growth of 5 mm or greater were treated with either translumbar glue or coil embolization of the lumbar source, or transarterial coil embolization of the inferior mesenteric artery. RESULTS: Type II endoleaks were detected in 90 (18.5%) patients. With a mean follow-up of 21.7 +/- 16 months, only 35 (7.2%) patients had type II endoleak that persisted for 6 months or longer. Aneurysm sac enlargement was noted in 5 patients, representing 1% of the total series. All 5 patients underwent successful translumbar sac embolization (n = 4) or transarterial inferior mesenteric artery embolization (n = 4) at a mean follow-up of 18.2 +/- 8.0 months, with no recurrence or aneurysm sac growth. No patient with treated or untreated type II endoleak has had rupture of the aneurysm. The mean global cost for treatment of persistent type II endoleak associated with aneurysm sac growth was US dollars 6695.50 (hospital cost plus physician reimbursement). Treatment in the 30 patients with persistent type II endoleak but no aneurysm sac growth would have represented an additional cost of US dollars 200000 or more. The presence or absence of a type II endoleak did not affect survival (78% vs 73%) at 48 months. CONCLUSIONS: Selective intervention to treat type II endoleak that persists for 6 months and is associated with aneurysm enlargement seems to be both safe and cost-effective. Longer follow-up will determine whether this conservative approach to management of type II endoleak is the standard of care. PMID- 14743130 TI - Preoperative selective intercostal angiography in patients undergoing thoracoabdominal aneurysm repair. AB - OBJECTIVE: This study was designed to test the hypothesis that detection of the location of the major artery supplying the spinal cord, that is, the artery of Adamkiewicz or the great radicular artery (GRA), with angiography would help prevent paraplegia. Knowing which intercostal artery provides this important branch would enable prompt, focused revascularization. METHOD: The surgical outcome in 131 patients with Crawford extent 1 and 2 degenerative aneurysms and 69 patients with descending thoracic aortic dissection was correlated with findings on selective intercostal arteriograms. Angiographic maneuvers were done with care, and the procedures were aborted if there was loose or "shaggy" mural thrombus, significant tortuosity, or difficulty entering each dissection channel. No attempts were made to find major contributions proximal to T6. Subarachnoid drains were placed in all patients, and all but five patients underwent distal aortic perfusion with controlled cooling to 32 degrees F. Five patients underwent cold circulatory arrest, enabling replacement of the distal aortic arch. We defined paraplegia simply as the inability to walk at hospital discharge, paraparesis as impaired ambulation, and both as having spinal cord dysfunction (SCD). RESULTS: A GRA was found in 65 (43%)of the 151 patients studied. Of the 65 patients with the GRA identified, SCD developed in 3 (4.6%) patients. Thirteen of 135 (9.6%) patients in whom the GRA was not identified, either because they were not studied or were studied and the GRA was not found, developed SCD (P =.35) However, when the GRA was identified, SCD occurred only in the group with aortic dissection. None of the 45 patients with degenerative aneurysms with the GRA identified had SCD, compared with 9 of 55 (16%) patients studied but without a GRA found (P =.01). CONCLUSION: The approach with selective intercostal angiography did not improve overall results. One third of our patients were not studied, and they fared as well as patients who were studied and the GRA was localized (not studied, 4 of 49, 8% with SCD; GRA localized, 3 of 65, 5% with SCD; P =.8). However, when the GRA was found, SCD occurred only in patients with aortic dissection. The studies confirmed the concept that the existence of mural thrombus in degenerative aneurysms results in the occlusion of many intercostal arteries, leaving those remaining patent to supply rich vascular watersheds through acquired collateral channels. As a result, in the group of patients with degenerative aneurysms, the identification of the critical intercostal artery allows focused reimplantation with uniform success. This is not the case in patients with aortic dissection. In those patients, most intercostal vessels remain patent, such that the insertion of one pair is insufficient to supply the paravertebral plexus and the spinal cord. Finally, failure to identify the GRA angiographically with our methods does not provide assurance that the GRA does not exist. Therefore negative findings did not provide license to ligate all intercostal arteries. PMID- 14743131 TI - Preservation of renal function with surgical revascularization in patients with atherosclerotic renovascular disease. AB - OBJECTIVE: Clinical and anatomic factors predictive of a favorable response to renal revascularization performed for renal function salvage remain poorly defined. To clarify decision making in such patients we reviewed a contemporary experience of surgical renal artery revascularization (RAR) performed primarily for preservation of renal function. METHODS: Between June 1990 and March 2001 (ensuring 1 year minimum follow-up), 96 patients with renal insufficiency (serum creatinine [Cr] concentration >or=1.5 mg/dL) and hypertension underwent RAR for preservation of renal function. Study end points included early and late renal function response, progression to dialysis dependence, and long-term survival. Variables potentially associated with these end points were assessed with univariate analysis, Cox regression analysis, and logistic regression analysis, and survival was assessed with the Kaplan-Meier method. RESULTS: Perioperative failure of RAR occurred in 3 patients (3%), with perioperative mortality in 4 patients (4.1%); thus 92 patients were available for long-term follow-up (mean, 39 months). Mean patient age was 70 +/- 9 years with a mean baseline Cr of 2.6 mg/dL (range, 1.5-7.8 mg/dL). Operative management consisted of aortorenal bypass in 38% of patients, extraanatomic bypass in 38% of patients, and endarterectomy in 24% of patients; 32% of patients required combined aortic revascularization and RAR, and 27% underwent bilateral RAR. At hospital discharge renal function had improved (20% decrement in Cr) in 41 (43%) patients, including 7 patients who were removed from dialysis; remained unchanged in 40 (41%) patients; and declined (20% increase in Cr) in 15 (16%) patients. At last follow-up renal function was either improved or unchanged in 72% of patients. Predictors of a favorable response to RAR at last follow-up included stable Cr at hospital discharge (odds ratio [OR], 7.1; 95% confidence interval [CI], 2.5-21.8; P =.0004) and decreased Cr at hospital discharge (OR, 16; 95% CI, 1.6-307.8; P <.0001); bilateral renal artery repair (OR, 3.1; 95% CI, 0.9-10.6; P =.07) approached clinical significance. Predictors of worsened excretory function at last follow-up included decline of renal function at hospital discharge (OR, 28.9; 95% CI, 5.0 165.4; P =.0002), intervention to treat unilateral renal artery stenosis (OR, 3.8; 95% CI, 0.8-16.6; P =.05), and level of baseline Cr (OR, 3.0; 95% CI, 1.0 4.0; P =.04). Progression to dialysis occurred in 16 (17%) patients. Dialysis free survival at 5 years was 50%, and overall actuarial survival at 5 years was 59%. Predictors of progression to dialysis during follow-up included treatment of complete renal artery occlusion (OR, 6.2; 95% CI, 1.3-29.5; P =.02), early failure of RAR (OR, 3.1; 95% CI, 0.7-14.2; P =.04) and baseline Cr (OR, 2.9; 95% CI, 1.3-6.1; P =.006). CONCLUSION: Long-term clinical success in the preservation of renal function, noted in 70% of patients, can be predicted by the initial response to RAR and by anatomic factors, in particular, bilateral repair. While extreme (mean Cr >or=3.2 mg/dL) renal dysfunction generally is predictive of poor long-term outcome, a subset of patients will experience favorable results, even to the extent of rescue from dialysis. These results may facilitate clinical decision making in the application of RAR for renal function salvage. PMID- 14743132 TI - A new look at outcomes after infrainguinal bypass surgery: traditional reporting standards systematically underestimate the expenditure of effort required to attain limb salvage. AB - BACKGROUND: Graft patency, limb salvage, and mortality are the traditional means of assessing the outcome of infrainguinal bypass surgery (IBS). However, these measures underestimate patient morbidity and fail to consider the entire spectrum of treatment required to restore the patients to their premorbid state. The aim of this study was to quantify the efforts required to achieve limb salvage by assessing three nontraditional outcomes: (1). index limb reoperation rate in 3 months, (2). hospital readmission rate in the first 6 months after IBS, and (3). wound-healing time. METHODS: We retrospectively analyzed 318 IBSs performed at a single institution. Repeat operations for limb or graft-related problems and readmissions within 6 months of the initial operation were recorded. When available, wound-healing time was determined. Pertinent demographics and comorbidities were subjected to univariate and multivariate analysis to determine risk factors for adverse outcomes. RESULTS: Seventy-two percent of patients underwent IBS for critical limb ischemia (CLI), and 84% had below-knee popliteal or distal bypasses. Among those who underwent IBS for CLI, 48.9% of patients required at least one reoperation within 3 months. Within 6 months, 49.3% of patients required hospital readmission. Time to heal exceeded 3 months in 54% of patients. After multivariate analysis, tissue loss and minority status were significant risk factors for reoperation within 3 months. Tissue loss and renal failure increased the odds for readmission within 6 months. Diabetes was the sole risk factor for prolonged wound healing. CONCLUSIONS: IBS for limb salvage is often complicated by prolonged recovery and multiple reoperations and readmissions. Traditional reporting standards for limb salvage operations need modification to reflect the true outcome of such procedures. PMID- 14743133 TI - Risk factors for autogenous infrainguinal bypass occlusion in patients with prosthetic inflow grafts. AB - OBJECTIVE: In patients with prosthetic inflow (PI) grafts the proximal anastomosis of autogenous infrainguinal bypass (AIB) can be placed on the PI or on a distal native vessel in the groin. This study was performed to determine the effect of placement of an AIB proximal anastomotic site in a patient with ipsilateral PI. METHODS: Patients undergoing AIB and PI between January 1990 and July 2002 were included in the study. They were classified into two groups on the basis of location of the proximal anastomosis. In group 1 the AIB proximal anastomosis was placed on the PI in the groin, whereas in group 2 the AIB proximal anastomosis was placed on a distal native groin artery. Patency, limb salvage, and patient survival in the two groups were calculated with the Kaplan Meier method. The Cox proportional hazards model was used to determine independent risk factors affecting AIB patency. RESULTS: Two hundred twenty-nine patients underwent AIB and PI. In group 1, 23 AIBs became thrombosed concurrent with 26 PI occlusions, and in group 2, 7 AIBs became thrombosed concurrent with 36 PI occlusions (P <.001). Five-year assisted primary patency, limb salvage, and patient survival in groups 1 and 2 were 50% and 75% (P <.001, log-rank test), 78% and 90% (P =.005, log-rank test), and 56% and 69% (P = NS, log-rank test), respectively. Factors independently associated with AIB occlusion are hypertension (hazard ratio [HR], 3.41; 95% confidence interval [CI], 1.65-7.05; P =.001), postoperative warfarin sodium therapy (HR, 1.86; 95% CI, 1.07-3.23; P =.03), continued smoking (HR, 1.72; 95% CI, 0.93-3.18; P =.08), AIB arising from PI (HR, 2.38; 95% CI, 1.35-4.18; P =.003), and PI occlusion (HR, 3.70; 95% CI, 2.15-6.36; P <.001). CONCLUSION: A proximal AIB anastomosis located directly on the PI is an independent risk factor for decreased AIB patency of equal or greater importance than current smoking, hypertension, or PI occlusion. The proximal anastomosis of an AIB in a patient with an ipsilateral PI should be placed on a distal native artery. PMID- 14743134 TI - Hemodynamic outcome of endovascular therapy for TransAtlantic InterSociety Consensus type B femoropopliteal arterial occlusive lesions. AB - OBJECTIVE: Endovascular therapy for moderate femoropopliteal arterial occlusive disease remains controversial. This study reviewed our experience with endovascular therapy for TransAtlantic InterSociety Consensus (TASC) type B disease, which is defined as multiple stenoses less than 3 cm in diameter or a single stenosis or occlusion 3 to 5 cm in diameter. Stenosis-free patency was used as an objective end point to evaluate the hemodynamic outcome. METHODS: A retrospective review was performed of all patients who had undergone endovascular treatment of TASC type B lesions between 1997 and 2002 at two referral centers. Balloon angioplasty was performed in all patients. Stenting was used selectively as an adjunct in patients with suboptimal angioplasty results. The treated sites were examined with duplex ultrasound scanning at 6-month to 12-month intervals. Stenosis-free patency was defined as the absence of stenosis greater than 50% diameter in the treated arterial segment with standard duplex criteria. RESULTS: One hundred fifteen limbs in 98 patients were studied. The indication for treatment was claudication in 92 patients (80%) and ischemic rest pain or gangrene in the remaining patients (20%). Multiple lesions greater than 3 cm were treated in 89 limbs (77%), and a single lesion 3 to 5 cm long was treated in 26 limbs (23%). Balloon angioplasty alone was performed in 74 limbs (65%), and angioplasty and adjunctive stenting was performed in 41 limbs (35%). Endovascular therapy was technically successful in all but one patient (99%), and there was no perioperative mortality or limb loss. During follow-up recurrent stenosis was detected in 46 limbs (40%), and reocclusion occurred in 11 limbs (10%). Seven patients (6%) underwent surgical bypass after endovascular treatment failure. The aggregate 1-year stenosis-free patency rate at life table analysis was 55.1%. The 1-year stenosis-free patency for angioplasty alone was 58%, compared with 51% (NS) for angioplasty and adjunctive stenting. Univariate regression analysis failed to demonstrate a difference in stenosis-free patency for demographic variables, medical comorbidities, and anatomic characteristics (multiple vs single lesions; number of angioplasty procedures). CONCLUSIONS: Endovascular therapy for TASC type B femoropopliteal lesions is safe and technically feasible. However, the length of time that a treated arterial segment remains free of stenosis is limited, and is not improved with adjunctive stenting. Recurrent stenosis, not occlusion, was the most common study end point, and few patients subsequently required surgical bypass. Predictors of outcome after endovascular therapy for TASC type B lesions were not identified in this study. PMID- 14743135 TI - Family history is a major determinant of subclinical peripheral arterial disease in young adults. AB - BACKGROUND: Cardiovascular disease is prevalent in first-degree relatives of young adults with premature-onset peripheral arterial occlusive disease (premature PAD), but it is not known whether the genetic influence is independent of other risk factors, the most prevalent of which is smoking. This study was performed to determine the relative contributions of family history and smoking to the development of occult PAD in siblings of patients with premature PAD. METHODS: The prevalence of occult PAD was determined with carotid, abdominal, and lower extremity duplex ultrasound scanning in 50 asymptomatic siblings (25 men, 25 women) of patients with premature PAD (onset 20 cigarettes/day) and 50 nonsmokers. RESULTS: Duplex ultrasound scans demonstrated raised arterial lesions in 20 asymptomatic siblings (40%), 20 heavy smokers (40%), and 2 nonsmoking control subjects (4%), resulting in 42 affected and 108 unaffected subjects. As expected, smokers were far more likely to have arterial lesions than were nonsmokers (odds ratio [OR], 11.19; 95% confidence interval [CI], 4.1-30.7; P <.0001). Individuals with a family history of premature PAD were almost three times more likely to have arterial lesions than those with no family history of cardiovascular disease (OR, 2.76; 95% CI, 1.3-5.8; P =.006). Stratified and multivariable logistic regression indicated no interaction between smoking and family history, indicating simple additive effects on PAD. CONCLUSIONS: Family history is a major determinant of occult PAD in young adults, and is at least as important as standard atherosclerotic risk factors. A high proportion of heavy smokers have clearly detectable lesions even though asymptomatic. Smoking and family history act additively to increase the risk for premature PAD. PMID- 14743136 TI - Patients undergoing infrainguinal bypass to treat atherosclerotic vascular disease are underprescribed cardioprotective medications: effect on graft patency, limb salvage, and mortality. AB - INTRODUCTION: Established American Heart Association guidelines recommend the use of statin drugs, angiotensin converting enzyme (ACE) inhibitors, and antiplatelet agents in patients with systemic atherosclerosis, such as those undergoing operative intervention to treat peripheral atherosclerotic disease. Many of these patients have not received treatment of coronary heart disease and have not been prescribed these medications. Whether statin drugs and ACE inhibitors confer an improvement in graft patency, limb salvage, and operative mortality is unknown. METHODS: Consecutive patients who underwent infrainguinal bypass between 1997 and 2002 were evaluated for demographic data, comorbid disease, medication use, as well as cumulative graft patency, limb salvage, and mortality. Univariate, multivariate logistical regression, and Kaplan-Meier analyses were performed. P <.05 was considered significant. RESULTS: Two hundred ninety-three patients (mean age, 64 years; 67% men) underwent 338 infrainguinal bypass procedures with autologous vein (n = 218), prosthetic grafts (n = 88), or composite prosthetic vein grafts (n = 32). Limb salvage was the operative indication in 75% of procedures. Coexisting diseases included hypertension (70%), diabetes (52%), hyperlipidemia (37%), coronary heart disease (51%), congestive heart failure (14%), and active tobacco use (30%). Statin drugs were taken by 56% of patients, ACE inhibitors by 54% of patients, and antiplatelet agents or warfarin sodium (Coumadin) by 93% of patients. Postoperative graft surveillance was done in 39% of patients. Cumulative graft patency was 73%, limb salvage was 85%, and mortality was 9%, with a mean follow-up of 17 months. Factors independently associated with increased graft patency included statin drug use (odds ratio [OR], 3.7; 95% confidence interval [CI], 2.1-6.4), male sex (OR, 2.8; 95% CI, 1.6 5.0), and graft surveillance (OR, 2.4; 95% CI, 1.3-4.5). Factors associated with decreased amputation rate were statin drug use (OR, 0.34; 95% CI, 6.15-0.77) and graft surveillance (OR, 0.23; 95% CI, 6.1-0.63). Factors associated with decreased mortality included graft surveillance (OR, 0.18; 95% CI, 0.1-0.56), whereas congestive heart failure (OR, 6.5; 95% CI, 2.5-17) and hemodialysis dependent renal failure (OR, 29; 95% CI, 6.1-140) were associated with increased mortality. Kaplan-Meier analysis showed that only ACE inhibitors were associated with lower mortality (P =.05) CONCLUSIONS: Patients undergoing infrainguinal bypass are undertreated with respect to cardioprotective medications. ACE inhibitor use is associated with lower mortality, and statin drug use is associated with improved graft patency and limb salvage. Institution of consensus guidelines concerning these medications should be considered by all vascular specialists, including vascular surgeons. PMID- 14743137 TI - Reliability of extracranial carotid artery duplex ultrasound scanning: value of vascular laboratory accreditation. AB - OBJECTIVES: The purpose of this study was to evaluate the reliability of carotid duplex ultrasound scanning performed by nonaccredited vascular laboratories and to assess the clinical effect on patient management. METHODS: We retrospectively reviewed concordance of findings of carotid duplex ultrasound scanning between laboratories accredited by the Intersocietal Commission for Accreditation of Vascular Laboratories and nonaccredited laboratories in 174 patients with asymptomatic disease referred to tertiary care community hospitals for surgical evaluation for carotid endarterectomy (CEA) between January 2001 and December 2002, and evaluated changes in clinical management made on the basis of repeat examinations. RESULTS: Concordant findings were noted in 171 of 348 arteries (49%), predominantly those with minimal or mild disease (114 arteries; 67%). Discordant findings of no clinical significance were found in 54 arteries (16%). Clinically significant discordant findings were noted in 123 arteries (35%) in 107 patients (61%). In 104 arteries (88 patients) stenosis was overestimated by the nonaccredited laboratory secondary to technical error (19 arteries), use of B mode imaging data alone (36 arteries), and use of inappropriate velocity criteria (49 arteries). None of these patients underwent CEA. Stenosis was significantly underestimated in 19 arteries (19 patients); all of these patients underwent uncomplicated CEA. CONCLUSIONS: Incorrect physician interpretation of data is the most common cause of error in carotid duplex ultrasound scanning performed in nonaccredited vascular laboratories. Results of carotid duplex ultrasound scanning from nonaccredited laboratories should be considered with extreme caution, and do not appear reliable in planning treatment of obstructive disease. PMID- 14743139 TI - Multistate improvement in process and outcomes of carotid endarterectomy. AB - OBJECTIVES: The purpose of this study was to assess the effect of community-wide performance measurement and feedback on key processes and outcomes of carotid endarterectomy (CEA). METHODS: Complete medical record (hospital chart) review for indications, care processes, and outcomes was performed on a random sample of Medicare patients undergoing CEA in 10 states (Arkansas, Georgia, Illinois, Indiana, Iowa, Kentucky, Michigan, Nebraska, Ohio, Oklahoma) during baseline (Jun 1, 1995 to May 31, 1996) and remeasurement (Jun 1, 1998 to May 31, 1999) periods. In addition to review of the index hospital stay, hospital admissions within 30 days of the procedure were reviewed and the Medicare enrollment database queried to identify out-of-hospital deaths, to determine 30-day outcome results. The baseline data by state were provided to the Medicare Quality Improvement Organizations (QIOs) in the respective states, and quality improvement initiatives were encouraged. RESULTS: We reviewed 9945 primary CEA alone procedures, 236 CEA and coronary artery bypass grafting (CABG) procedures, and 380 repeat CEA operations during the baseline period (B), and 9745 primary CEA alone procedures, 233 CEA and CABG procedures, and 401 repeat CEA operations during the remeasurement period (R). There was a significant decrease in the combined event rate (30-day stroke or mortality) for CEA alone procedures between baseline and remeasurement (B, 5.6%; R, 5.0%). A decrease occurred in each of the indication strata; transient ischemic attack or stroke (B, 7.7%; R, 6.9%), nonspecific symptoms (B, 5.9%; R, 5.4%), and no symptoms (B, 4.1%; R, 3.8%). The combined event rate also decreased for CEA and CABG (B, 17.4%; R, 13.3%) and repeat CEA operations (B, 6.8%; R, 5.7%). The remeasurement period state-to-state variation in combined event rate for CEA alone ranged from 2.7% (Georgia) to 5.9% (Indiana) for all indications combined, from 4.4% (Georgia) to 10.9% (Michigan) in patients with recent transient ischemia or stroke, from 1.4% (Georgia) to 6.0% (Oklahoma) in patients with no symptoms, and from 3.7% (Georgia) to 7.9% (Indiana) in patients with nonspecific symptoms. There were significant increases in preoperative antiplatelet administration (62%-67%; P <.0001) and patching (29% 45%; P =.05) from baseline to remeasurement in the CEA alone subset. Preoperative antiplatelet administration and patching were associated with improved outcomes in the combined baseline and remeasurement data. CONCLUSIONS: Community-wide quality improvement initiatives with performance measurement and confidential reporting of provider level data can lead to improvement in important care processes and outcomes. There is considerable variation between states in outcome and process, and thus continued room for improvement. Quality improvement projects that include standardized confidential outcome reporting should be encouraged. Preoperative antiplatelet therapy administration and patching rates should be considered as evidence-based performance measures. PMID- 14743140 TI - Prosthetic thigh arteriovenous access: outcome with SVS/AAVS reporting standards. AB - PURPOSE: Differences in the reporting methods of results for arteriovenous (AV) access can dramatically affect apparent outcome. To enable meaningful comparisons in the literature, the Society for Vascular Surgery and the American Association for Vascular Surgery (SVS/AAVS) recently published reporting standards for dialysis access. The purpose of the present study was to determine infection rates, patency rates, and possible predictive factors for prosthetic thigh AV access outcomes with the reporting standards of the SVS/AAVS. METHODS: A retrospective analysis was performed of all patients who underwent placement of thigh AV access by the Surgical Teaching Service at Greenville Memorial Hospital between 1989 and 2001. Outcomes were determined based on SVS/AAVS Standards for Reports Dealing with AV Accesses. The rate of revision per year of access patency was also determined; this end point more accurately reflects the true cost and morbidity associated with AV access than do patency or infection rates alone. RESULTS: One hundred twenty-five polytetrafluoroethylene thigh AV accesses were placed in 100 patients. Nine accesses were excluded from the study, six because there was no patient follow-up and 3 as a result of deaths unrelated to the access procedure and which occurred less than 30 days after access placement. There were six (4%) late access-related deaths. There were 18 (15%) early access failures, related to infection in 14 cases (12%), thrombosis in three cases (2%), and steal in one case (1%). Early failure was more common in patients with diabetes mellitus (P =.036). The primary and secondary functional patency rates were 19% and 54%, respectively, at 2 years. Infection occurred in 48 (41%) accesses. The patency and infection rates were not influenced by patient age, gender, body mass index, or diabetes mellitus. The median number of interventions per year of access patency was 1.68, and this outcome was positively correlated with body mass index (P <.001). CONCLUSIONS: Prosthetic AV access in the thigh is associated with higher morbidity compared with that reported for the upper extremity, and should be considered only if no upper extremity AV access option is available. Early access failure and the requirement for an increased number of interventions to reestablish and maintain access patency are more common in patients with diabetes mellitus and obesity. The number of interventions per year of access patency is a valuable end point when assessing the outcome of AV access procedures. PMID- 14743141 TI - Deep vein harvest: predicting need for fasciotomy. AB - OBJECTIVE: Deep thigh veins, including the superficial femoral, superficial femoropopliteal, and profunda femoris veins, are versatile autogenous conduits for arterial reconstruction. Although late venous complications are unusual, deep vein harvest may induce severe venous hypertension and predispose the limb to acute compartment syndrome. The purpose of this study was to define the frequency of fasciotomy in patients undergoing deep vein harvest and to identify clinical predictors of the need for fasciotomy after deep vein harvest. METHODS: Over 9 years, 162 patients underwent arterial reconstruction with deep vein harvested from 264 limbs. Indications for deep vein harvest included aortofemoral reconstruction in 127 patients, brachiocephalic arterial reconstruction in 22 patients, and visceral arterial reconstruction in 13 patients. RESULTS: Fasciotomy was performed in 47 of 264 limbs (17.8%) after deep vein harvest. The prevalence of fasciotomy after deep vein harvest was 20.6% for patients requiring aortofemoral reconstruction, whereas no patients underwent fasciotomy after deep vein harvest for mesenteric or brachiocephalic arterial reconstruction (P =.0068). Fasciotomy was performed in 20.7% of limbs after complete deep vein harvest to a level below the adductor hiatus, but no fasciotomies were performed in patients undergoing subtotal deep vein harvest, ending above the adductor hiatus (P =.0023). The mean preoperative ankle-brachial index (ABI) was significantly lower in limbs requiring fasciotomy (ABI, 0.39 +/- 0.05), compared with patients who did not require fasciotomy (ABI, 0.79 +/- 0.02; P <.0001). Fasciotomy was performed in 76.0% of limbs undergoing concurrent ipsilateral greater saphenous vein (GSV) and deep vein harvest, compared with 11.7% of patients undergoing deep vein harvest alone (P <.0001). The mean volume of intraoperative fluid administered to patients requiring fasciotomy was almost 50% higher than the fluid resuscitation received by patients who did not require fasciotomy (9.6 +/- 1.2 L vs 6.5 +/- 0.6 L; P <.0001). Logistic regression analysis determined that lower preoperative ABI (odds ratio [OR], 60.1; 95% confidence interval [CI], 12.5-289.3; P <.0001) and concurrent harvest of the ipsilateral GSV (OR, 9.9; 95% CI, 3.1-31.3; P <.0001) were predictors of the need for fasciotomy. CONCLUSIONS: One in four patients undergoing deep vein harvest for aortofemoral reconstruction may be expected to develop acute compartment syndrome and require fasciotomy. The risk appears to be greatest in patients with severe lower extremity ischemia and in patients undergoing simultaneous GSV and deep vein harvest. Prophylactic fasciotomy may be appropriate in patients with both risk factors, but vigilance for the development of compartment syndrome after deep vein harvest is required in all patients undergoing deep vein harvest for aortofemoral reconstruction. PMID- 14743142 TI - System to decrease length of stay for vascular surgery. AB - OBJECTIVES: Reduction of length of stay (LOS) is critical for optimal use of hospital resources. We developed and evaluated a system to aggressively reduce LOS for vascular surgery. METHOD: Key to this system, which we introduced on January 1, 2001, was appointment of a LOS officer, who communicated daily during hospitalization with patients and families about discharge planning, organized outpatient services for wound care and rehabilitation to transition patients quickly to nonhospital care, and had biweekly meetings with relevant paramedical services. LOS for 509 patients operated on in 2000 (standard group) was compared with LOS for 474 operated on in 2001 and 595 patients operated on in 2002 (LOS reduction groups). Data for all patients with aortic aneurysm, carotid artery stenosis, lower extremity critical ischemia or amputation, and foot debridement were included. RESULTS: LOS in 2000 averaged 8.5 days, compared with 5.9 days in 2001 and 5.6 days in 2002. All decreases in LOS for each diagnostic category in 2001 and 2002 were statistically significant (P = <.001-.03). There was no significant increase in readmission rate (2.2% vs 1.9% and 2.0%, respectively), mortality rate (0.8% vs 0.6% and 0.7%, respectively), or percent of patients who received endovascular treatment (18% vs 16% and 14%, respectively). These decreases in LOS saved the hospital more than US dollars 616200 in 2001, and US dollars 847550 in 2002 (US dollars 500/patient-day). CONCLUSIONS: A committed LOS officer with major specific daily responsibilities for decreasing LOS and discharging patients resulted in a 31% to 33% decrease in LOS, with important cost savings to the hospital and no negative effect on patient care. PMID- 14743143 TI - Ruptured inflammatory abdominal aortic aneurysm: insights in clinical management and outcome. AB - BACKGROUND: Ruptured inflammatory abdominal aortic aneurysm (AAA) is relatively rare, and little has been written on the outcome of operative treatment. METHODS: Patients undergoing attempted repair of ruptured inflammatory AAA between 1995 and 2001 were included in a retrospective case-cohort study. Demographic, clinical, and operative factors were analyzed, together with in-hospital morbidity, in-hospital mortality, and duration of postoperative hospital stay. RESULTS: Of 297 patients who underwent attempted operative repair of ruptured AAA, 24 (8%) had an inflammatory aneurysm. Twenty-two patients were men, and two were women; median age was 69 years (range, 51-85 years). Operative findings revealed a contained hematoma in 16 patients (70%), free rupture in 3 patients (13%), aortocaval fistula in 4 patients (17%), and aortoenteric fistula in 1 patient (4%). Of 273 noninflammatory ruptured AAAs, only 2 AAA (1%) were associated with primary aortic fistula. Ten patients (42%) with inflammatory AAA died in hospital, compared with 117 of 273 patients (43%) without inflammation. Median postoperative stay was 10 days (range, 0-35 days). Of the 14 patients with inflammatory lesions who survived, 11 had postoperative complications; 4 patients had acute renal failure, three of whom required temporary renal replacement therapy. CONCLUSIONS: Ruptured inflammatory AAA is associated with a higher incidence of aortic fistula than is ruptured noninflammatory AAA. Repair of ruptured inflammatory AAA is not associated with increased operative mortality compared with repair of ruptured noninflammatory AAA. PMID- 14743144 TI - Hypogastric artery bypass to preserve pelvic circulation: improved outcome after endovascular abdominal aortic aneurysm repair. AB - OBJECTIVE: This study was carried out to compare the functional outcomes after hypogastric artery bypass and coil embolization for management of common iliac artery aneurysms in the endovascular repair of aortoiliac aneurysms (EVAR). METHODS: Between 1996 and 2002, 265 patients underwent elective or emergent EVAR. Data were retrospectively reviewed for 21 (8%) patients with iliac artery aneurysms 25 mm or larger that involved the iliac bifurcation. Patients underwent hypogastric artery bypass (n = 9) or coil embolization (n = 12). Interviews about past and current levels of activity were conducted. A disability score (DS) was quantitatively graded on a discrete scale ranging from 0 to 10, corresponding to "virtually bed-bound" to exercise tolerance "greater than a mile." Worsening or improvement of symptoms was expressed as a difference in DS between two time points (-, worsening; +, improvement). RESULTS: There was no difference in age (72.6 +/- 7.3 years vs 73.1 +/- 6.4 years), sex (male-female ratio, 8:1 vs 11:1), abdominal aortic aneurysm size (60.1 +/- 5.9 mm vs 59.3 +/- 7.0 mm), or number of preoperative comorbid conditions (1.9 +/- 0.8 vs 2.1 +/- 0.8) between hypogastric bypass and coil embolization groups, respectively. Mean follow-up was shorter after hypogastric bypass (14.8 vs 20.5 months; P <.05). There was no difference in the mean overall baseline DS between the bypass and the embolization groups (8.0 vs 7.8). Six (50%) of the 12 patients with coil embolization reported symptoms of buttock claudication ipsilateral to the occluded hypogastric artery. No symptoms of buttock claudication were reported after hypogastric bypass (P <.05). There was a decrease in the DS after both procedures; however, coil embolization was associated with a significantly worse DS compared with hypogastric artery bypass (4.5 vs 7.3; P <.001). In 4 (67%) of 6 patients with claudication after coil embolization symptoms improved, with a DS of 5.4 at last follow-up. This was significantly worse than in patients undergoing hypogastric artery bypass, with a DS of 7.8 at last follow-up (P <.001). There was no difference between the groups in duration of procedure, blood loss, length of hospital stay, morbidity, or mortality (0%). CONCLUSIONS: Hypogastric artery bypass to preserve pelvic circulation is safe, and significantly decreases the risk for buttock claudication. Preservation of pelvic circulation results in significant improvement in the ambulatory status of patients with common iliac artery aneurysms, compared with coil embolization. PMID- 14743145 TI - Accelerated progression of carotid stenosis in patients with previous external neck irradiation. AB - OBJECTIVE: Radiotherapy to the head and neck often results in carotid stenosis, but the course of disease is unknown. We investigated the natural history and progression of asymptomatic carotid stenosis induced by external irradiation. Patients and methods The study included 130 carotid arteries in 95 patients who had received external radiation therapy to the head and neck area and who had asymptomatic, mild internal carotid artery or common carotid artery stenosis. Stenosis of 15% to 49% on duplex ultrasound (US) scans defined mild (<50%) disease. Another 95 arteries in 74 patients with matched degree of carotid artery stenosis but who had not received radiation therapy were used as control. Both groups were followed up prospectively with serial duplex US scanning, and degree of carotid artery stenosis was categorized as 15% to 49%, 50% to 69%, 70% to 99%, and occlusion. Progression of carotid artery stenosis was defined as increase in stenosis from less than 50% to 50% or greater at ultrasonography. Secondary end points included progression to higher disease category, new cerebrovascular symptoms, and death. Data from irradiated arteries was compared with control data with the life table method. A Cox regression model was used to analyze disease progression, adjusted for covariates of sex, age, smoking, diabetes, and hypertension. RESULTS: Mean follow-up was 36 months. Adjusted freedom from progression rates at 3 years were 65% for irradiated arteries and 87% for control arteries at life-table analysis (P =.035; odds ratio, 3.1). The annualized progression rate from less than 50% to 50% or greater in irradiated arteries was 15.4%, compared with 4.8% in nonirradiated arteries. A long history of cervical irradiation (>6 years) was the only significant risk factor for disease progression. There was no difference between the two groups regarding development of new symptoms or mortality. CONCLUSIONS: Carotid stenosis associated with external irradiation progresses more rapidly compared with nonirradiated atherosclerotic arteries. Aggressive surveillance is recommended. PMID- 14743146 TI - Intraoperative carotid artery duplex scanning in a modern series of 650 consecutive primary endarterectomy procedures. AB - PURPOSE: Thromboembolic complications after carotid endarterectomy are frequently associated with technical defects. We analyzed the effect of intraoperative duplex scanning in detection of significant but clinically unsuspected technical defects and residual common carotid artery (CCA) disease as a potential source of postoperative transitory ischemic attack (TIA) and stroke. METHODS: From April 2000 to April 2003, 650 consecutive primary carotid endarterectomy procedures were performed in 590 patients at a single institution by two vascular surgeons. Patients included 335 men (57%) and 255 women (43%). Indications for surgery were asymptomatic internal carotid artery (ICA) stenosis (>or=70%) in 464 patients (71%). All procedures were performed with the patient under general anesthesia, with synthetic patch angioplasty in 644 (99.1%). Major technical defects at intraoperative duplex scanning (>30% luminal internal carotid artery stenosis, free-floating clot, dissection, arterial disruption with pseudoaneurysm) were repaired. CCA residual disease was reported as wall thickness (0.7-4.8 mm; mean, 1.7 +/- 0.7) and percent stenosis (16%-67%; mean, 32% +/- 8%) in all cases. Postoperative 30-day TIA, stroke, and death rates were analyzed. RESULTS: There were no clinically detectable postoperative thromboembolic events in this series. All 15 major defects (2.3%) identified with duplex scanning were successfully revised. These included 7 intimal flaps, 4 free-floating clots, 2 ICA stenoses, 1 ICA pseudoaneurysm, and 1 retrograde CCA dissection. Diameter reduction ranged from 40% to 90% (mean, 67 +/- 16%), and peak systolic velocity ranged from 69 to 497 cm/s (mean, 250 +/- 121 cm/s). Thirty-one patients (5%) with the highest residual wall thickness (>3mm) in the CCA and 19 (3%) with the highest CCA residual diameter reduction (>50%) did not have postoperative stroke or TIA. Overall postoperative stroke and mortality rates were 0.3% and 0.5%, respectively; combined stroke and mortality rate was 0.8%. One stroke was caused by hyperperfusion, and the other occurred as an extension of a previous cerebral infarct. No patients had TIAs. Two deaths were caused by myocardial infarction, and one death by respiratory insufficiency. CONCLUSION: We believe intraoperative duplex scanning had a major role in these improved results, because it enabled detection of clinically unsuspected significant lesions. Residual disease in the CCA does not seem to be a harbinger of stroke or TIA. PMID- 14743147 TI - Balloon angioplasty as a treatment of failing infrainguinal autologous vein bypass grafts. AB - PURPOSE: The purpose of this study was to evaluate the role of balloon angioplasty in the treatment of failing infrainguinal vein bypass (IVB) grafts. METHODS: A retrospective chart review of patients undergoing revision of a failing IVB graft by vascular surgeons at a tertiary care center from 1990 to 2001 was performed. Failing bypass grafts were identified by routine duplex scan surveillance and physical examination. The criteria for endovascular intervention varied on the basis of surgeon preferences and time period; factors considered when choosing balloon angioplasty included significant comorbidities that precluded operative intervention, the lack of adequate conduit for surgical revision, or poor accessibility of the stenotic lesion. Data recorded included demographic patient data, type of IVB graft, patency status, further procedures performed, and all complications and mortalities. Cumulative primary and assisted patency rates were calculated by using Kaplan-Meier life-table analysis. RESULTS: A total of 45 balloon angioplasties were performed in 36 patients. There were 36 angioplasties of vein bypass grafts, and additional balloon angioplasties were performed on nine of these patients. Locations of IVB grafts included femoropopliteal (13 patients), femorodistal (13), and popliteal to distal (10). Initial success was achieved in 33 of 36 vein bypass grafts (91.7%). In these bypass grafts, the stenotic lesions were identified and treated at the proximal anastomosis (3 patients), mid-bypass graft (6 patients), and distal anastomosis (27 patients). Autogenous vein was used for all bypass grafts. Cumulative vein bypass graft (life-table analysis) primary patency rates (those free of occlusion or bypass graft threatening stenosis) were 74.2% at 6 months, 62.7% at 12 months, and 58.2% at 24 months. Repeat interventions included surgical thrombectomy with vein patch angioplasty or bypass graft revision, as well as repeat balloon angioplasty with or without thrombolysis. Cumulative assisted vein bypass graft patency rates (those free of occlusion or bypass graft threatening stenosis) were 87.0%, 83.2%, and 78.9% at 6, 12, and 24 months, respectively. Two patients (4%) developed thigh hematomas; no other procedure-related complications were noted, and there were no deaths in the perioperative period. CONCLUSION: Balloon angioplasty of failing infrainguinal vein bypass grafts can be successfully performed with a low rate of complications. Acceptable short-term patency can be achieved. This procedure should be considered as an initial option in failing IVB grafts. PMID- 14743148 TI - Freedom from secondary interventions to treat stenotic disease after percutaneous transluminal angioplasty of infrarenal aorta: long-term results. AB - PURPOSE: The purpose of this study was to determine the long-term results (1-15 years) of percutaneous transluminal angioplasty (PTA) of localized atherosclerotic lesions of the infrarenal aorta. METHODS: This was a retrospective study. From January 1987 to January 2002, 69 patients underwent PTA of an isolated stenosis of the lower abdominal aorta under local anesthesia in the department of interventional radiology. All atherosclerotic lesions were hemodynamically significant, defined as a subjective report of walking distance less than one block, resting pain, or trophic changes in combination with diameter reduction of 50% or greater at duplex ultrasound scanning and angiography. RESULTS: The female-male ratio of study patients was 3.6:1; mean age was 58 years. Endovascular treatment was initially technically and clinically successful in all but one patient (98%), who had a near total occlusion. No major complications were noted. Mean follow-up was 57 months (range, 6 months-15 years). At life table analysis, 5-year primary patency was 75%, and secondary patency was 97%. Twelve patients (17%) required repeat interventions because of hemodynamically significant recurrent stenosis in combination with severe clinical symptoms. Most recurrent stenoses (67%) were successfully treated with repeat endovascular procedures. CONCLUSIONS: Early and long-term results of PTA (with additional stent placement) of isolated stenosis of the infrarenal aorta are good. This minimally invasive procedure is a true alternative to traditional surgical methods. PMID- 14743149 TI - Matrix metalloproteinase inhibition reduces intimal hyperplasia in a porcine arteriovenous-graft model. AB - BACKGROUND: The patency of arteriovenous (AV) polytetrafluoroethylene grafts for hemodialysis is impaired by intimal hyperplasia (IH) at the venous outflow tract. IH mainly consists of vascular smooth muscle cells, fibroblasts, and extracellular matrix proteins. Because matrix metalloproteinases (MMPs) are enzymes able to degrade extracellular matrix proteins such as elastin and collagen and also stimulate migration of vascular smooth muscle cells, we hypothesized that BB2983 (a broad-spectrum MMP inhibitor) could reduce IH in AV grafts. METHODS: In 12 pigs, AV grafts were created bilaterally between the carotid artery and the jugular vein. Six pigs received the oral MMP inhibitor (MMPi), and six pigs served as a control. Four weeks after AV shunting, the grafts and adjacent vessels were excised and underwent histologic analysis. Quantification of elastin content was performed on Elastin von Gieson-stained sections. RESULTS: At the venous outflow tract, IH was strongly inhibited after MMPi when compared with the control group (1.02 +/- 0.26 mm(2) vs 2.14 +/- 0.38 mm(2); P =.027). The medial area did not differ significantly. In the control group elastin density decreased compared with nonoperated veins. This decrease was not observed in the MMPi group (nonoperated, 6.3% +/- 0.4%; MMPi, 7.2% +/- 0.7% vs untreated, 3.6% +/- 0.5%; P =.0004). Outward remodeling of the vein was not influenced by MMP inhibition. CONCLUSION: MMPi reduces IH formation at the venous outflow tract of AV grafts in pigs, probably by inhibiting elastin degradation. These data suggest that MMP inhibitors might be useful for minimizing IH in AV grafts, thus prolonging patency rates of AV grafts in patients on hemodialysis. PMID- 14743150 TI - Concomitant blockade of platelet-derived growth factor receptors alpha and beta induces intimal atrophy in baboon PTFE grafts. AB - OBJECTIVE: Although current treatments for restenosis attempt to prevent the development of intimal hyperplasia, an alternative strategy is to induce intimal atrophy after restenosis has developed. Because platelet-derived growth factor (PDGF) is a smooth muscle cell growth and survival factor, we tested the hypothesis that complete blockade of PDGF by using antibodies against PDGF receptors alpha and beta would cause intimal atrophy in a baboon vascular graft model. METHODS: We administered chimeric antibodies against PDGF receptor alpha or PDGF receptor beta, either separately or together, to baboons with bilateral prosthetic aortoiliac grafts, the intimas of which had reached maximal size before treatment was begun. High blood flow, which we have previously shown to cause intimal atrophy, was induced through one graft to serve as a positive control. After 2 weeks, the intima lining the grafts was assessed for cross sectional area, cell proliferation, and apoptosis by standard morphologic and immunohistochemical techniques. RESULTS: Blocking both PDGF receptors simultaneously reduced the cross-sectional area of the normal-flow graft intima by 44% (P <.05 vs control), whereas treatment with the individual antibodies did not significantly alter intimal area. Blockade of both receptors also inhibited smooth muscle cell proliferation by 66% (P <.05 vs control), whereas neither antibody alone altered proliferation. In contrast, all treatments increased smooth muscle cell apoptosis threefold to fivefold. CONCLUSIONS: These data suggest that simultaneous inhibition of cell proliferation and stimulation of cell death by the administration of antibodies to both PDGF receptor alpha and receptor beta is required for intimal atrophy in this baboon graft model. In addition, these data provide an in vivo model for the pharmacologic induction of intimal atrophy and introduce a novel clinical approach to treat intimal hyperplasia. Clinical relevance This study introduces the concept of pharmacologic induction of intimal atrophy. Intimal hyperplasia plagues all forms of arterial reconstruction. Currently, the only effective treatment of these restenotic lesions is balloon angioplasty or operative revision. An alternative approach to patients with clinically significant intimal hyperplasia might be to stimulate intimal regression by modulating growth and survival factors required for intimal maintenance. Although PDGF is known to be critical in intimal formation, the results of this study suggest that PDGF is also critical for intimal maintenance. Inhibition of the PDGF system may prove to be a clinically applicable approach for inducing intimal atrophy. PMID- 14743151 TI - Matrix metalloproteinase 2 is suppressed by trapidil, a CD40-CD40 ligand pathway inhibitor, in human abdominal aortic aneurysm wall. AB - BACKGROUND: The activation of inflammatory cells and the production of matrix metalloproteinases (MMPs) are important in the pathogenesis of abdominal aortic aneurysm (AAA). Previous studies have demonstrated that the antiplatelet agent trapidil has multiple actions, including suppression of MMP expression through the inhibition of the CD40-CD40 ligand (CD40-CD40L) pathway in cultured cells. A recent clinical study suggested that trapidil might have functions beyond its antiplatelet action. Methods and results In the present study, we performed immunohistochemical analysis and semiquantitative reverse transcription polymerase chain reaction to evaluate the effect of trapidil on the production of MMPs in cultured aortic tissues from patients with infrarenal AAA (n = 9) and control patients with aortoiliac occlusive disease (n = 7). The tissue concentrations of both MMP-2 and MMP-9 were significantly higher in AAA walls than in control aortic walls. Both trapidil and an anti-CD154 (CD40L) antibody significantly suppressed the protein production and mRNA expression of MMP-2 but did not inhibit those of MMP-9 in organ cultures of AAA wall specimens. MMP-9 was produced by macrophages and a lot of neutrophils in AAA tissues, whereas MMP-2 was derived from macrophages. CD40 was expressed on macrophages but not on neutrophils, and this expression could explain the differential effect of trapidil on the production of MMP-2 and MMP-9. CONCLUSIONS: Trapidil, a CD40 CD40L pathway inhibitor, suppressed mRNA expression and protein production of MMP 2 in AAA tissues, suggesting a potential therapeutic approach for the prevention or treatment of AAA. PMID- 14743152 TI - Retrograde visceral vessel revascularization followed by endovascular aneurysm exclusion as an alternative to open surgical repair of thoracoabdominal aortic aneurysm. AB - Staged visceral artery revascularization with occlusion of the proximal lumen enables endovascular exclusion of the entire thoracoabdominal aneurysm from a femoral approach. This technique has been successfully used in three patients at high risk for conventional repair. PMID- 14743153 TI - Aneurysm of superior mesenteric vein: case report with 5-year follow-up and review of the literature. AB - Venous aneurysms are less common than arterial aneurysms in clinical practice, and the occurrence of isolated cases is a topic for publication. Aneurysms of the superior mesenteric vein are rare, and their origin is unknown. Many aneurysms are asymptomatic, and the diagnosis is established from radiologic findings. Others are diagnosed after complications such as gastrointestinal bleeding or thrombosis with associated abdominal pain. Because of the rarity of this disease and consequent absence of standard treatment, therapy must be adapted to fit each case. We present a case report of an aneurysm of the superior mesenteric vein. The diagnosis of this anomaly was made after investigation of abdominal pain. Computed tomography (CT) scans demonstrated the mass. Clinical treatment was administered, and no aneurysm growth was observed after 5 years of follow-up. PMID- 14743154 TI - Isolated contralateral vertebral artery injury in a stab wound to the neck. AB - Ipsilateral vertebral artery injuries are a rare entity following trauma to the neck. We discuss the first case of an isolated right vertebral artery injury in a patient with a left stab wound. The patient required bilateral neck explorations and, due to massive bleeding, a median sternotomy in order to obtain control of the proximal segment of the right vertebral artery. We emphasize the importance for trauma surgeons to be familiar with basic but important vascular exposures. The anatomy, surgical exposure, and management of these rare injuries are discussed. PMID- 14743155 TI - Intravenous leiomyomatosis: two cases with different routes of tumor extension. AB - Intravenous leiomyomatosis is a rare smooth muscle tumor. We report two cases of intravenous leiomyomatosis that grew along different routes of the venous system into the inferior vena cava and the right atrium. The different route of extension makes a difference in the ease of excision of tumor masses. Using MEDLINE together with the references in each publication, we identified all cases of intracardiac leiomyomatosis reported in the English literature in the period between 1980 and 2003 and performed a brief review on this potentially lethal disease entity. PMID- 14743156 TI - Endovascular external-to-internal iliac bypass as an adjunct to endovascular aneurysm repair for patients with extensive common iliac artery aneurysmal disease. PMID- 14743157 TI - Videoscopic approach to femoral bifurcation. AB - Femoral artery exposure can lead to local complications, which can compromise the postoperative course of effective arterial reconstruction. We report a new technique of vascular reconstruction in which a videoscopic approach to the femoral bifurcation is used. To our knowledge, the videoscopic approach to the femoral bifurcation has not been previously described. The main advantages of this technique include a limited cutaneous incision and avoidance of skinfolds, especially in obese patients. PMID- 14743158 TI - Mandate for creation of a national peripheral arterial disease public awareness program: an opportunity to improve cardiovascular health. AB - BACKGROUND: There has been increasing recognition of the detrimental effect of peripheral arterial disease (PAD) on the health of Americans, and yet there is no common national program of public PAD education designed to diminish this effect. FORMAT: To heighten awareness of this problem, a 2-day PAD Public Education Strategy Meeting was recently attended by representatives of 17 professional societies and public health associations whose missions support the prevention, diagnosis, treatment, and rehabilitation of vascular diseases. This Public Education Strategy Meeting was intended to provide the rationale and structure to create a national PAD public awareness campaign to diminish the health effect of PAD and to improve cardiovascular outcomes in the United States. This document (1). provides the rationale for creation of a national PAD public education program; (2). reviews the development and success of national hypertension, hypercholesterolemia, and Women's Heart Health public education programs as models of educational efficacy; (3). elucidates how the work of many vascular professionals has led to a national consensus for creation of a national PAD public educational program; (4). provides an overview of the National Heart, Lung, and Blood Institute PAD education meeting; and (5). outlines the "next steps" required to accomplish these goals. OUTCOME: This meeting achieved consensus that we share responsibility for developing accurate, unified messages to promote PAD awareness and improved care. Participants agreed that the creation of such messages should be linked to plans to disseminate them to all Americans at risk. A consensus was reached that such messages, when commonly created and offered to the public, are most likely to achieve the rewards in better health that all Americans deserve. The Vascular Disease Foundation, a not-for-profit foundation whose mission includes public education about the prevention, diagnosis, treatment, and rehabilitation of PAD, will devote its resources to guide a new coalition in this process and to create a national PAD awareness campaign. During 2003 and 2004, the coalition will create the organizational underpinnings and time line for what will undoubtedly be a multiyear effort. CONCLUSION: Participants of the Public Education Strategy Meeting agreed to create a broad coalition to develop a National PAD Public Awareness Program, with the objectives to develop and disseminate public education massages on PAD. A successful national PAD education program will contribute to creation of a broader mandate to improve global cardiovascular health in the United States. PMID- 14743159 TI - Ethics of introducing new operating room technology. PMID- 14743181 TI - Disaster and terrorism preparedness: what pediatricians need to know. PMID- 14743183 TI - Molecular characterization of bipolar disorder by comparing gene expression profiles of postmortem brains of major mental disorders. AB - We performed the oligonucleotide microarray analysis in bipolar disorder, major depression, schizophrenia, and control subjects using postmortem prefrontal cortices provided by the Stanley Foundation Brain Collection. By comparing the gene expression profiles of similar but distinctive mental disorders, we explored the uniqueness of bipolar disorder and its similarity to other mental disorders at the molecular level. Notably, most of the altered gene expressions in each disease were not shared by one another, suggesting the molecular distinctiveness of these mental disorders. We found a tendency of downregulation of the genes encoding receptor, channels or transporters, and upregulation of the genes encoding stress response proteins or molecular chaperons in bipolar disorder. Altered expressions in bipolar disorder shared by other mental disorders mainly consisted of upregulation of the genes encoding proteins for transcription or translation. The genes identified in this study would be useful for the understanding of the pathophysiology of bipolar disorder, as well as the common pathophysiological background in major mental disorders at the molecular level. In addition, we found the altered expression of LIM and HSPF1 both in the brains and lymphoblastoid cells in bipolar disorder. These genes may have pathophysiological importance and would be novel candidate genes for bipolar disorder. PMID- 14743184 TI - In search of a depressed mouse: utility of models for studying depression-related behavior in genetically modified mice. AB - The ability to modify mice genetically has been one of the major breakthroughs in modern medical science affecting every discipline including psychiatry. It is hoped that the application of such technologies will result in the identification of novel targets for the treatment of diseases such as depression and to gain a better understanding of the molecular pathophysiological mechanisms that are regulated by current clinically effective antidepressant medications. The advent of these tools has resulted in the need to adopt, refine and develop mouse specific models for analyses of depression-like behavior or behavioral patterns modulated by antidepressants. In this review, we will focus on the utility of current models (eg forced swim test, tail suspension test, olfactory bulbectomy, learned helplessness, chronic mild stress, drug-withdrawal-induced anhedonia) and research strategies aimed at investigating novel targets relevant to depression in the mouse. We will focus on key questions that are considered relevant for examining the utility of such models. Further, we describe other avenues of research that may give clues as to whether indeed a genetically modified animal has alterations relevant to clinical depression. We suggest that it is prudent and most appropriate to use convergent tests that draw on different antidepressant-related endophenotypes, and complimentary physiological analyses in order to provide a program of information concerning whether a given phenotype is functionally relevant to depression-related pathology. PMID- 14743185 TI - St John's wort and imipramine-induced gene expression profiles identify cellular functions relevant to antidepressant action and novel pharmacogenetic candidates for the phenotype of antidepressant treatment response. AB - Both the prototypic tricyclic antidepressant imipramine (IMI) and the herbal product St John's wort (SJW) can be effective in the treatment of major depressive disorder. We studied hypothalamic gene expression in rats treated with SJW or IMI to test the hypothesis that chronic antidepressant treatment by various classes of drugs results in shared patterns of gene expression that may underlie their therapeutic effects. Individual hypothalami were hybridized to individual Affymetrix chips; we studied three arrays per group treatment. We constructed 95% confidence intervals for expression fold change for genes present in at least one treatment condition and we considered genes to be differentially expressed if they had a confidence interval excluding 1 (or -1) and had absolute difference in expression value of 10 or greater. SJW treatment differentially regulated 66 genes and expression sequence tags (ESTs) and IMI treatment differentially regulated 74 genes and ESTs. We found six common transcripts in response to both treatments. The likelihood of this occurring by chance is 1.14 x 10(-23). These transcripts are relevant to two molecular machines, namely the ribosomes and microtubules, and one cellular organelle, the mitochondria. Both treatments also affected different genes that are part of the same cell function processes, such as glycolytic pathways and synaptic function. We identified single-nucleotide polymorphisms in the human orthologs of genes regulated both treatments, as those genes may be novel candidates for pharmacogenetic studies. Our data support the hypothesis that chronic antidepressant treatment by drugs of various classes may result in a common, final pathway of changes in gene expression in a discrete brain region. PMID- 14743186 TI - Transmission disequilibrium test (TDT) for case-control studies. AB - Genetic association, case-control studies are becoming a major instrument in the attempt to identify disease susceptibility markers of complex diseases. However, a major drawback of population-based studies of genetic association is the confounding effect of the population subdivision. We developed a statistic named T-value that estimates the differential transmission of marker alleles from heterozygous parents to the affected offspring, based on population data. Our method does not assume Hardy-Weinberg equilibrium and it can be used in very different population structures. A great advantage of this approach is that the genetic structure of the population can be assessed with a few unlinked loci and using classical population genetics theory (i.e. Wright's F-statistics). Four general models, assuming either one population with random mating, or one population without random mating, or several populations with random mating within them, or several populations without random mating within them, were developed to determine the behavior of the T-value under different mating conditions. Although a complete knowledge of the population structure is ideal to choose the best model, the simulations show that for a total inbreeding of 0.30 or less the last three models gave very similar estimates of the T-value. The model that assumed that total departure of Hardy-Weinberg proportions is due to population subdivision was the most robust under different scenarios of population structure. In sum, this study describes a novel procedure that can be used to identify the transmission of disease susceptibility markers in population based studies. PMID- 14743188 TI - Variations in 1 alpha,25-dihydroxyvitamin D3 and 25-hydroxyvitamin D3 serum levels during allogeneic bone marrow transplantation. PMID- 14743189 TI - Unrelated cord blood transplantation in an infant with severe multisystem Langerhans cell histiocytosis: clinical outcome, engraftment and culture of monocyte-derived dendritic cells. PMID- 14743190 TI - Extracorporeal photochemotherapy for treatment of fasciitis in chronic graft versus-host disease. PMID- 14743191 TI - Correction of persistent thrombocytopenia by a boost of CD133+ selected stem cells in a patient transplanted for Wiskott-Aldrich syndrome 10 years ago. PMID- 14743193 TI - Acute pancreatitis due to tacrolimus in a case of allogeneic bone marrow transplantation. PMID- 14743192 TI - Results of an unrelated transplant search strategy using partially HLA-mismatched cord blood as an immediate alternative to HLA-matched bone marrow. AB - Cord blood (CB) is an alternative to other sources of stem cells for transplantation. However, the impact of including CB in the initial strategy of unrelated graft search in a cohort of patients has been the object of limited analysis. Here, we report the results of such a strategy in 91 consecutive children. Absence of mismatch was required for adult donors, and up to two mismatches were allowed for CB grafts, with a nucleated cell dose over 2.5 x 10(7) cells/kg. A graft was found for 84 of the 85 children who remained available for a 3-month search. In all, 64 patients were transplanted, 36 with CB and 28 with bone marrow (BM). Primary graft failure, acute grade II-IV and extensive chronic graft-versus-host disease occurred in five, five and zero CB, and in three, one and two BM patients, respectively. The 3-year survival was 59% in CB and 57% in BM patients. Accepting CB as a source of stem cells offers a graft to almost every child in need of an unrelated transplantation, with a probability of survival similar to that of unrelated BM transplantation. PMID- 14743194 TI - Contemporaneous appearance, 18 years after allogeneic bone marrow transplantation, of myelodysplastic syndrome in the patient and the donor. AB - We report the case of the development of two different stages of the same clonal disorder in two patients sharing the same bone marrow due to a previous bone marrow allotransplant. The transplanted patient developed severe aplasia with myeloid blasts, different from those of the previously cured leukemia. Chimerism evaluated by microsatellite analyses confirmed a full donor phenotype. At the same time, the donor of the bone marrow transplantation developed a refractory anemia with excess blasts. We speculate on the presence of an undetectable pre existing pathological clone in the transplanted bone marrow, which have evolved in the two patients. PMID- 14743195 TI - How long after neutrophil recovery should myeloid growth factors be continued in autologous hematopoietic stem cell transplant recipients? AB - Growth factors are routinely used after autotransplantation to accelerate hematopoietic recovery, and are continued until the absolute neutrophil count (ANC) is >/=0.5 x 10(9)/l on 3 consecutive days. Since ANC often increases to very high levels with this strategy, we discontinued growth factor on the first day ANC reached 0.5 x 10(9)/l in 45 patients (Study Group), and compared their subsequent ANC to 108 historic controls who received growth factor longer. While ANC on the day after reaching 0.5 x 10(9)/l was comparable between groups, ANC on the third day was significantly higher in the Control Group (2.3 vs 4.9 x 10(9)/l; P=0.0003). When compared to the first day, ANC in the Study Group was higher by a median of 140% on the third day and by 450% in the Control Group (P=0.0002). A significantly higher proportion of patients experienced a decline in ANC after the first day in the Study Group. However, only one patient in the Study Group became neutropenic transiently and ANC recovered spontaneously the next day. The incidence of fever and hospitalization were comparable. We conclude that growth factors can be discontinued after autotransplantation the day the ANC reaches 0.5 x 10(9)/l, without compromising neutrophil recovery. PMID- 14743196 TI - Bacteremia after hematopoietic stem cell transplantation: incidence and predictive value of surveillance cultures. AB - We studied 622 transplants undertaken between 1982 and 2001 to: (1) determine the incidence, timing and etiology of bacteremias, and (2) examine the ability of routine surveillance cultures to predict bacteremias. A total of 404 episodes (0.65 episode per patient) occurred in 248 patients, due to coagulase-negative staphylococci (n=171, 42%), Gram-negative bacteria (n=129, 32%), streptococci (n=48, 12%), other Gram-positive bacteria (n=33, 8%), anaerobes (n=9, 2%) and fungi (n=14, 3%). Bacteremias were more frequent in allogeneic (0.96 episode/patient) compared to autologous (0.44) transplants (P<0.0001). The overall incidence decreased from 0.92 episode/patient until 1990 to 0.66 in 1991 1996 and 0.55 in 1997-2001 (P<0.0001), but this was only observed in autologous transplants. Among them, 212 (53%) occurred before hospital discharge and 192 (47%) thereafter. This proportion was lower for coagulase-negative staphylococci, other Gram-positive bacteria and Gram-negative bacteria compared to other agents (P=0.001). In 50% of the cases, the agent responsible for the bacteremic episode was present in routine surveillance cultures previously. IN CONCLUSION: (1) bacteremias remain a frequent complication, particularly in allogeneic transplantation, even long after hospital discharge; (2) routine surveillance cultures can predict bacteremias in 50% of the cases, but the practical impact of this observation is limited in view of the costs. PMID- 14743197 TI - Peripheral blood stem cell mobilization following CHOP plus rituximab therapy combined with G-CSF in patients with B-cell non-Hodgkin's lymphoma. AB - We mobilized peripheral blood stem cells (PBSC) following CHOP plus rituximab (CHOP-R) therapy, and compared with the findings following CHOP therapy without rituximab. All patients were given G-CSF starting from day 11 after CHOP therapy. Patients in the CHOP-R group (n=8) were given rituximab on day 12. Target CD34(+) cells number was collected in a single leukapheresis on day 14, from all the eight patients in the CHOP-R group. PBSC mobilization kinetics, CD34(+) cells yield and colony-forming ability in the graft collection, toxicity during mobilization, and engraftment after transplantation of CHOP-R group were not significantly different from those in the CHOP group (n=8). In all patients given CHOP-R therapy, CD20(+) cells and immunoglobulin heavy chain (IgH) rearrangement in the graft collection were undetectable by flow-cytometric analysis and Southern blot analysis, respectively, but with PCR analysis two of eight grafts were positive for IgH rearrangement. While further studies are needed to evaluate the efficacy of purging and the outcome of patients undergoing autologous transplantation, CHOP-R therapy can be safely and effectively used in the mobilization phase of PBSC collection, without excess clinical toxicity or deleterious effect on PBSC mobilization kinetics or engraftment time. PMID- 14743198 TI - Effect of peripheral blood progenitor cell dose on hematopoietic recovery: identification of minimal progenitor cell requirements for rapid engraftment. AB - Repeated high-dose chemotherapy (HDC) with stem cell support is advocated for curative treatment of epithelial ovarian cancer patients, requiring large quantities of progenitor cell harvest. Although the switchover to peripheral blood stem cell transplantation has generally made possible the harvest of large quantities of progenitor cells, the minimum threshold is still pertinent for planning the safe conduct of HDC. However, as the minimum threshold for safe peripheral blood stem cell transplantation (PBSCT) is not yet established, this study was designed to clarify the minimum amount of progenitor cells required for prompt recovery of hematopoietic. Retrospective analysis was performed on 52 HDCs administered in 37 ovarian cancer patients. After autologous bone marrow aspiration (10 patients) or peripheral blood stem cell harvest (27 patients), colony-forming unit granulocyte macrophage (CFU-GM) were enumerated prior to cryopreservation. Numbers of CFU-GM were again calculated before reinfusion and the patients were divided into eight groups: 0.13-<0.4, 0.4-<0.7, 0.7-<1.0, 1.0 <3.5, 3.5-<5.0, 5.0-<10.0, 10.0-<20.0 and >20.0 (x 10(5)/kg). The minimum CFU-GM threshold (x 10(5)/kg) was found to be 1.0-<3.5 for platelets and 3.5-<5.0 for white blood cells. Higher infusion doses did not lead to significant benefits in hematopoietic reconstruction. These results indicate that preservation of a minimum of 7-10 x 10(5)/kg CFU-GM is recommended for the safe conduct of tandem HDCs. PMID- 14743199 TI - Chronic graft-versus-host disease: where do we go from here? AB - Chronic graft-versus-host disease (GvHD) remains a significant cause of late morbidity and mortality following allogeneic stem cell transplantation. However, patients with chronic GvHD are very heterogeneous, making evaluation and treatment difficult. Corticosteroids remain the most effective primary treatment of this condition. Randomized trials have not confirmed the beneficial effect of additional cyclosporine, even in patients with higher risk features. For patients failing initial therapy, no standard therapy is available. A plethora of drugs have been reported to have activity and promise in this disease. However, the majority of reports are small retrospective studies, with few prospective trials. The marked variability in the reported response rates for many of these novel agents highlights a number of problems in the evaluation and management of chronic GvHD. In addition to the heterogeneity of patients, there are no uniform definitions for treatment failure, prognostic criteria to stratify patients according to risk, or to evaluate response to treatment, which in many cases is largely subjective. The challenge ahead is to develop more uniform criteria for defining many of these important variables, which is likely to lead to the design of better prospective clinical trials to improve the outcome of patients with this condition. PMID- 14743200 TI - Impaired bone marrow hematopoietic progenitor cell function in rheumatoid arthritis patients candidated to autologous hematopoietic stem cell transplantation. AB - We have evaluated bone marrow morphology, percentage of bone marrow CD34(+) cells, proliferative activity of bone marrow precursors, clonogenic assay (BFU-E and CFU-GM) in short-term bone marrow cultures, and bone marrow cell apoptosis, together with serum TNF-alpha and IL-6, in 16 chronic, refractory RA patients, as well as in five healthy controls. Of 16 RA patients (68.7%), 11 showed a reduced bone marrow cellularity, while it was normal in all the controls. In RA patients, the median percentage of CD34(+) bone marrow cells, the median percentage of proliferating bone marrow myeloid precursors, and the median number of both BFU-E and CFU-GM colonies were significantly lower than observed in the controls. As far as TNF-alpha and IL-6 titers is concerned, the latter did not significantly differ from controls' values, while TNF-alpha titers were significantly lower in healthy controls. Finally, the median apoptotic index of early bone marrow myeloid cells of RA patients was significantly higher compared with controls. These observations may identify the biological risk factors for impaired mobilization and/or engraftment when RA patients are candidates for autologous hematopoietic stem cell grafting. PMID- 14743201 TI - Outbreaks of infectious diseases in stem cell transplant units: a silent cause of death for patients and transplant programmes. AB - Following the closure of the National Blood and Bone Marrow Transplant Unit in Dublin, because of an outbreak of vancomycin-resistant enterococcal infection, a survey was carried out by the EBMT to investigate the occurrence of outbreaks of infection in SCT units and the impact on patient morbidity, mortality and the administration of the transplant programme over a 10-year period from 1991 to 2001. A total of 13 centres reported 23 outbreaks of infection involving 231 patients: 10 bacterial, eight viral and five fungal outbreaks were reported and 56 deaths were attributed to infection. All fungal and bacterial deaths and the majority of viral deaths occurred in allograft recipients. In all outbreaks, the infection was reported to be hospital acquired and in all the viral, and half the bacterial infections, cross-infection was a major factor. All viral, four of 10 bacterial and three of five fungal outbreaks occurred in HEPA filtered rooms. A total of 12 SCT units reported a partial or total closure. The introduction of mandatory quality management systems such as JACIE should result in a change in attitude to 'incident reporting' and together with future surveys should reduce the incidence of infectious outbreaks in SCT units. PMID- 14743202 TI - Disruption of cell-type-specific methylation at the Maspin gene promoter is frequently involved in undifferentiated thyroid cancers. AB - Cancer-associated DNA hypomethylation is as prevalent as cancer-linked hypermethylation, but the biological significance of DNA hypomethylation in carcinogenesis is less understood. The expression of Maspin (mammary serpin) in differentiated normal cells is regulated by epigenetic modifications in a cell type-specific manner. Paradoxical Maspin expression due to epigenetic modification has been addressed in several cancer cell types. To elucidate the role of the Maspin gene in thyroid cancer, we studied methylation status in the promoter region and its expression in six human undifferentiated thyroid cancer cell lines and in specimens from 92 primary thyroid tumors, consisting of six follicular adenomas, 56 well-differentiated thyroid cancers (WDTCs), 17 poorly differentiated thyroid cancers (PDTCs) and 13 undifferentiated thyroid cancers (UDTCs). Three of the six cell lines overexpressed Maspin mRNA and its protein product, but the remaining three did not. The methylation status at the promoter region was inversely correlated with Maspin expression. In Maspin-negative cell lines, Maspin expression was induced by treatment with 5-aza-2'-deoxycytidine, a DNA demethylating agent. Immunoreactivity for Maspin protein was frequently detected in UDTCs (8/13, 62%) and PDTCs (7/17, 41%). Immunoreactivity for Maspin was diffusely positive in UDTCs, and was restricted to dedifferentiated components of the tumor in PDTCs. Positive immunoreactivity was infrequent in WDTCs (1/56, 2%), and all follicular adenomas and normal thyroid glands were completely negative. Their methylation status evaluated by the methylation specific PCR method showed a good inverse correlation with their immunoreactivity in surgically resected specimens. Our data suggest that overexpression of Maspin by DNA hypomethylation is closely associated with morphological dedifferentiation in thyroid cancers. PMID- 14743203 TI - Identification and validation of an ERBB2 gene expression signature in breast cancers. AB - ERBB2 is a transmembrane tyrosine kinase receptor encoded by a gene located in chromosome region 17q12. Overexpression of ERBB2, generally by way of gene amplification, plays a role in mammary oncogenesis. This alteration can be overcome by use of the humanized monoclonal antibody trastuzumab (Herceptin). Accurate determination of ERBB2 status is required for appropriate use of this targeted therapy and is currently analysed by immunohistochemistry (IHC) on tissue sections and/or fluorescence in situ hybridisation (FISH) on interphase chromosomes. We have studied the gene expression profiles of a series of 213 breast tumours and 16 breast cancer cell lines with known ERBB2 status, using Ipsogen's DiscoveryChip microarrays with approximately 9000 cDNAs. We have identified 36 genes and expressed sequence tags that were differentially expressed in tumours and in cell lines with and without ERBB2 protein overexpression. This ERBB2-specific gene expression signature (GES) contained 29 overexpressed genes including the ERBB2 gene itself, five genes located in its immediate vicinity on 17q12, non-17q genes such as GATA4 and eight downregulated genes including oestrogen receptor alpha (ER). Some correlations were validated at the protein level using IHC on tissue microarrays. The GES was able to distinguish ERBB2-negative and -positive cancer samples, as well as FISH-negative and FISH-positive ERBB2 2+ IHC samples. PMID- 14743204 TI - p53 protects from replication-associated DNA double-strand breaks in mammalian cells. AB - Genetic instability caused by mutations in the p53 gene is generally thought to be due to a loss of the DNA damage response that controls checkpoint functions and apoptosis. Cells with mutant p53 exhibit high levels of homologous recombination (HR). This could be an indirect consequence of the loss of DNA damage response or p53 could have a direct role in HR. Here, we report that p53-/ mouse embryonic fibroblasts (MEFs) exhibit higher levels of the RAD51 protein and increased level of spontaneous RAD51 foci Agents that stall replication forks, for example, hydroxyurea (HU), potently induce HR repair and RAD51 foci. To test if the increase in RAD51 foci in p53-/- MEFs was due to an increased level of damage during replication, we measured the formation of DNA double strand breaks (DSBs) in p53+/+ and p53-/- MEFs following treatments with HU. We found that HU induced DSBs only in p53-/- MEFs, indicating that p53 is involved in a pathway to protect stalled replication forks from being collapsed into a substrate for HR. Also, p53 is upregulated in response to agents that inhibit DNA replication, which supports our hypothesis. Finally, we observed that the DSBs produced in p53-/- MEFs did not result in a permanent arrest of replication and that they were repaired. Altogether, we suggest that the effect of p53 on HR and RAD51 levels and foci can be explained by the idea that p53 suppresses formation of recombinogenic lesions. PMID- 14743205 TI - Frequent loss of RUNX3 gene expression in human bile duct and pancreatic cancer cell lines. AB - RUNX3, a Runt domain transcription factor involved in TGF-beta signaling, is a candidate tumor-suppressor gene localized in 1p36, a region commonly deleted in a wide variety of human tumors, including those of the stomach, bile duct, and pancreas. Recently, frequent inactivation of RUNX3 has been demonstrated in human gastric carcinomas. In this study, to examine the involvement of RUNX3 abnormalities in tumorigenesis of bile duct as well as pancreatic cancers, we investigated not only the expression but also methylation status of RUNX3 in 10 human bile duct and 12 pancreatic cancer cell lines. Seven (70%) of the bile duct and nine (75%) of the pancreatic cancer cell lines exhibited no expression of RUNX3 by both Northern blot analysis and the reverse transcriptase polymerase chain reaction. All of the 16 cell lines that did not express RUNX3 also showed methylation of the promoter CpG island of the gene, whereas the six cell lines that showed RUNX3 expression were not methylated or only partially methylated in the RUNX3 promoter region. Moreover, treatment with the methylation inhibitor 5' aza-2'-deoxycitidine activated RUNX3 mRNA expression in all of 16 cancer cell lines that originally lacked RUNX3 expression. Finally, hemizygous deletion of RUNX3, as detected by fluorescence in situ hybridization, was found in 15 of the 16 cancer cell lines that lacked RUNX3 expression. These data suggest that the inactivation of RUNX3 plays an important role in bile duct and pancreatic carcinogenesis, and that methylation is a common mechanism by which the gene is inactivated. PMID- 14743206 TI - Mutant p53 exerts a dominant negative effect by preventing wild-type p53 from binding to the promoter of its target genes. AB - Mutation of the p53 tumor suppressor gene is the most common genetic alteration in human cancer. A majority of these mutations are missense mutations in the DNA binding domain. As a result, the mutated p53 gene encodes a full-length protein incapable of transactivating its target genes. In addition to this loss of function, mutant p53 can have a dominant negative effect over wild-type p53 and/or gain of function activity independently of the wild-type protein. To better understand the nature of the tumorigenic activity of mutant p53, we have investigated the mechanism by which mutant p53 can exert a dominant negative effect. We have established several stable cell lines capable of inducibly expressing a p53 mutant alone, wild-type p53 alone, or both proteins concurrently. In this context, we have used chromatin immunoprecipitation to determine the ability of wild-type p53 to bind to its endogenous target genes in the presence of various p53 mutants. We have found that p53 missense mutants markedly reduce the binding of wild-type p53 to the p53 responsive element in the target genes of p21, MDM2, and PIG3. These findings correlate with the reduced ability of wild-type p53 in inducing these and other endogenous target genes and growth suppression in the presence of mutant p53. We also showed that mutant p53 suppresses the ability of wild-type p53 in inducing cell cycle arrest. This highlights the sensitivity and utility of the dual inducible expression system because in previous studies, p53-mediated cell cycle arrest is not affected by transiently overexpressed p53 mutants. Together, our data showed that mutant p53 exerts its dominant negative activity by abrogating the DNA binding, and subsequently the growth suppression, functions of wild-type p53. PMID- 14743207 TI - Phosphorylation status of c-Kit and Epo receptors, and the presence of wild-type p53 confer in vitro resistance of murine erythroleukemic cells to Celecoxib. AB - It is well established that selective COX-2 inhibitors exhibit potent effects against progression of select solid tumours. However, their effects on liquid tumours have not been fully established. By taking advantage of murine Friend Disease we have shown a strong antileukemic effect of celecoxib by determining novel in vitro targets. Western blot analyses revealed the expression of COX-2 in a panel of Friend Virus-transformed, splenic-derived primary erythroleukemic blasts and established cell lines generated in our laboratory. We have shown that celecoxib at concentrations as low as 20 microM significantly suppresses proliferation of the selected murine erythroleukemia cell line HB60-5. The greatest proliferative inhibition was seen at 40 microM of celecoxib, resulting in apoptosis. Our results also demonstrate that treatment of the established murine erythroleukemia cell line HB60-5 with celecoxib results in suppression of c-Kit and erythropoietin receptor (Epo-R) phosphorylation resulting in apoptosis, likely through decreased levels of survival factors. However, upon overexpression of c-Kit alone in these cells a significant increase in survival and twofold increase in proliferation in the presence of celecoxib were observed (P < 0.05). Finally, since responsiveness of our murine erythroleukemia cell lines to celecoxib is above the reported physiologically achievable levels in vivo, we have provided in vitro evidence to suggest that reduced sensitivity of erythroleukemic cells to lower doses of celecoxib may be a consequence of the loss of wild-type p53. These findings are pivotal in addressing potential discrepancies associated with sensitivity of murine erythroleukemic cells to celecoxib in vitro versus in vivo. PMID- 14743208 TI - Genomic alterations in spontaneous and carcinogen-induced murine melanoma cell lines. AB - We have conducted an analysis of genetic alterations in spontaneous murine melanoma cell line B16F0 and its two metastatic clones, B16F1 and B16F10 and the carcinogen-induced murine melanoma cell lines CM519, CM3205, and K1735. We found that unlike human melanomas, the murine melanoma cell lines did not have activating mutations in the Braf oncogene at exon 11 or 15. However, there were distinct patterns of alterations in the ras, Ink4a/Arf, and p53 genes in the two melanoma groups. In the spontaneous B16 melanoma cell lines, expression of p16Ink4a and p19Arf tumor suppressor proteins was lost as a consequence of a large deletion spanning Ink4a/Arf exons 1alpha, 1beta, and 2. In contrast, the carcinogen-induced melanoma cell lines expressed p16Ink4a but had inactivating mutations in either p19Arf (K1735) or p53 (CM519 and CM3205). Inactivation of p19Arf or p53 in carcinogen-induced melanomas was accompanied by constitutive activation of mitogen-activated protein kinases (MAPKs) and/or mutation associated activation of N-ras. These results indicate that genetic alterations in p16Ink4a/p19Arf, p53 and ras-MAPK pathways can cooperate in the development of murine melanoma. PMID- 14743209 TI - Frequent epigenetic inactivation of RASSF1A and BLU genes located within the critical 3p21.3 region in gliomas. AB - RASSF1A is a major tumor suppressor gene located at 3p21.3. We investigated the role of aberrant promoter region hypermethylation of RASSF1A in a large series of adult gliomas. RASSF1A was frequently methylated in both primary tumors (36/63; 57%) and tumor cell lines (7/7; 100%). Hypermethylation of RASSF1A in glioma cell lines correlated with loss of expression and treatment with a demethylating agent reactivated RASSF1A gene expression. Furthermore, re-expression of RASSF1A suppressed the growth of glioma cell line H4 in vitro. Next, we investigated whether other members of the RASSF gene family were also inactivated by methylation. NORE1B and RASSF3 were not methylated in gliomas, while NORE1A and RASSF5/AD037 demonstrated methylation in glioma cell lines but not in primary tumors. We then investigated the methylation status of three other candidate 3p21.3 tumor suppressor genes. CACNA2D2 and SEMA3B were not frequently methylated, but the BLU gene located just centromeric to RASSF1 was frequently methylated in glioma cell lines (7/7) and in 80% (35/44) of glioma tumors. In these tumor cell lines, BLU expression was restored after treatment with a demethylating agent. Loss of BLU gene expression in glioma tumors correlated with BLU methylation. There was no association between RASSF1A and BLU methylation. RASSF1A methylation increased with tumor grade, while BLU methylation was seen at similar frequencies in all grades. Our data implicate RASSF1A and BLU promoter methylation in the pathogenesis of adult gliomas, while other RASSF family members and CACNA2D2 and SEMA3B appear to have only minor roles. In addition, RASSF1A and BLU methylation appear to be independent and specific events and not due to region-wide changes in DNA methylation. PMID- 14743210 TI - Alterations in the p16(INK4a) and p53 tumor suppressor genes of hTERT immortalized human fibroblasts. AB - Exogenous expression of the catalytic subunit of telomerase, hTERT, in a normal human foreskin fibroblast cell strain resulted in telomerase activity and an extended proliferative lifespan prior to a period of crisis. Three immortalized cell lines with stably maintained telomere lengths were established from cells that escaped crisis. Each of these cultures underwent a significant downregulation of p16(INK4a) expression due to gene deletion events. One cell line also acquired mutations in both alleles of the p53 tumor suppressor gene. Downregulation of p16(INK4a) and loss of wild-type p53 expression occurred after escape from crisis, so these mutations are most likely not required for immortalization of these cells but rather were selected for during continuous growth in vitro. These findings emphasize the need for caution in the use of hTERT-immortalized cells in studies of normal cell biology or in tissue engineering and the need to monitor for genetic instability and the accumulation of mutations in both the p16(INK4a)/pRb and p53 pathways. PMID- 14743211 TI - Ultra-stable nanoparticles of CdSe revealed from mass spectrometry. AB - Nanoparticles under a few nanometres in size have structures and material functions that differ from the bulk because of their distinct geometrical shapes and strong quantum confinement. These qualities could lead to unique device applications. Our mass spectral analysis of CdSe nanoparticles reveals that (CdSe)(33) and (CdSe)(34) are extremely stable: with a simple solution method, they grow in preference to any other chemical compositions to produce macroscopic quantities. First-principles calculations predict that these are puckered (CdSe)(28)-cages, with four- and six-membered rings based on the highly symmetric octahedral analogues of fullerenes, accommodating either (CdSe)(5) or (CdSe)(6) inside to form a three-dimensional network with essentially heteropolar sp(3) bonding. This is in accordance with our X-ray and optical analyses. We have found similar mass spectra and atomic structures in CdS, CdTe, ZnS and ZnSe, demonstrating that mass-specified and macroscopically produced nanoparticles, which have been practically limited so far to elemental carbon, can now be extended to a vast variety of compound systems. PMID- 14743212 TI - High-aspect-ratio bulk micromachining of titanium. AB - Recent process developments have permitted the highly anisotropic bulk micromachining of titanium microelectromechanical systems (MEMS). By using the metal anisotropic reactive ion etching with oxidation (MARIO) process, arbitrarily high-aspect-ratio structures with straight sidewalls and micrometre scale features have been bulk micromachined into titanium substrates of various thicknesses, ranging from 0.5-mm sheet down to 10-microm free-standing titanium foils. Bulk micromachined structures are generally free of residual stresses and are preferred when large, rigid, flat and/or high-force actuators are desired. However, so far there has been a limited ability to select materials on the basis of specific application in bulk micromachining, primarily because of the predominance of MEMS processes dedicated to single-crystal silicon, such as silicon deep reactive ion etching. The MARIO process permits the creation of bulk titanium MEMS, which offers potential for the use of a set of material properties beyond those provided by traditional semiconductor-based MEMS. Consequently, the MARIO process enables the fabrication of novel devices that capitalize on these assets to yield enhanced functionalities that would not be possible with traditional micromechanical material systems. PMID- 14743213 TI - Remotely actuated polymer nanocomposites--stress-recovery of carbon-nanotube filled thermoplastic elastomers. AB - Stimuli-responsive (active) materials undergo large-scale shape or property changes in response to an external stimulus such as stress, temperature, light or pH. Technological uses range from durable, shape-recovery eye-glass frames, to temperature-sensitive switches, to the generation of stress to induce mechanical motion. Here, we demonstrate that the uniform dispersion of 1-5 vol.% of carbon nanotubes in a thermoplastic elastomer yields nanocomposites that can store and subsequently release, through remote means, up to 50% more recovery stress than the pristine resin. The anisotropic nanotubes increase the rubbery modulus by a factor of 2 to 5 (for 1-5 vol.%) and improve shape fixity by enhancing strain induced crystallization. Non-radiative decay of infrared photons absorbed by the nanotubes raises the internal temperature, melting strain-induced polymer crystallites (which act as physical crosslinks that secure the deformed shape) and remotely trigger the release of the stored strain energy. Comparable effects occur for electrically induced actuation associated with Joule heating of the matrix when a current is passed through the conductive percolative network of the nanotubes within the resin. This unique combination of properties, directly arising from the nanocomposite morphology, demonstrates new opportunities for the design and fabrication of stimuli-responsive polymers, which are otherwise not available in one material system. PMID- 14743214 TI - The formation of sp3 bonding in compressed BN. AB - Attributed to their specific atomic bonding, the soft, graphite-like, hexagonal boron nitride (h-BN) and its superhard, diamond-like, cubic polymorph (c-BN) are important technological materials with a wide range of applications. At high pressure and temperature, h-BN can directly transform to a hexagonal close-packed polymorph (w-BN) that can be partially quenched after releasing pressure. Previous theoretical calculations and experimental measurements (primarily on quenched samples) provided substantial information on the transition, but left unsettled questions due to the lack of in situ characterization at high pressures. Using inelastic X-ray scattering to probe the boron and nitrogen near K-edge spectroscopy, here we report the first observation of the conversion process of boron and nitrogen sp(2)- and p-bonding to sp(3) and the directional nature of the sp(3) bonding. In combination with in situ X-ray diffraction probe, we have further clarified the structure transformation mechanism. The present archetypal example opens two enormous, element-specific, research areas on high pressure bonding evolutions of boron and nitrogen; each of the two elements and their respective compounds have displayed a wealth of intriguing pressure-induced phenomena that result from bonding changes, including metallization, superconductivity, semiconductivity, polymerization and superhardness. PMID- 14743215 TI - Flexible active-matrix displays and shift registers based on solution-processed organic transistors. AB - At present, flexible displays are an important focus of research. Further development of large, flexible displays requires a cost-effective manufacturing process for the active-matrix backplane, which contains one transistor per pixel. One way to further reduce costs is to integrate (part of) the display drive circuitry, such as row shift registers, directly on the display substrate. Here, we demonstrate flexible active-matrix monochrome electrophoretic displays based on solution-processed organic transistors on 25-microm-thick polyimide substrates. The displays can be bent to a radius of 1 cm without significant loss in performance. Using the same process flow we prepared row shift registers. With 1,888 transistors, these are the largest organic integrated circuits reported to date. More importantly, the operating frequency of 5 kHz is sufficiently high to allow integration with the display operating at video speed. This work therefore represents a major step towards 'system-on-plastic'. PMID- 14743216 TI - A physical and functional map of the human TNF-alpha/NF-kappa B signal transduction pathway. AB - Signal transduction pathways are modular composites of functionally interdependent sets of proteins that act in a coordinated fashion to transform environmental information into a phenotypic response. The pro-inflammatory cytokine tumour necrosis factor (TNF)-alpha triggers a signalling cascade, converging on the activation of the transcription factor NF-kappa B, which forms the basis for numerous physiological and pathological processes. Here we report the mapping of a protein interaction network around 32 known and candidate TNF alpha/NF-kappa B pathway components by using an integrated approach comprising tandem affinity purification, liquid-chromatography tandem mass spectrometry, network analysis and directed functional perturbation studies using RNA interference. We identified 221 molecular associations and 80 previously unknown interactors, including 10 new functional modulators of the pathway. This systems approach provides significant insight into the logic of the TNF-alpha/NF-kappa B pathway and is generally applicable to other pathways relevant to human disease. PMID- 14743217 TI - Protein kinase D regulates basolateral membrane protein exit from trans-Golgi network. AB - Protein kinase D (PKD) binds to diacylglycerol (DAG) in the trans-Golgi network (TGN) and is activated by trimeric G-protein subunits beta gamma. This complex then regulates the formation of transport carriers in the TGN that traffic to the plasma membrane in non-polarized cells. Here we report specificity of different PKD isoforms in regulating protein trafficking from the TGN. Kinase-inactive forms of PKD1, PKD2 and PKD3 localize to the TGN in polarized and non-polarized cells. PKD activity is required only for the transport of proteins containing basolateral sorting information, and seems to be cargo specific. PMID- 14743218 TI - The tumour suppressor RASSF1A regulates mitosis by inhibiting the APC-Cdc20 complex. AB - The tumour suppressor gene RASSF1A is frequently silenced in lung cancer and other sporadic tumours as a result of hypermethylation of a CpG island in its promoter. However, the precise mechanism by which RASSF1A functions in cell cycle regulation and tumour suppression has remained unknown. Here we show that RASSF1A regulates the stability of mitotic cyclins and the timing of mitotic progression. RASSF1A localizes to microtubules during interphase and to centrosomes and the spindle during mitosis. The overexpression of RASSF1A induced stabilization of mitotic cyclins and mitotic arrest at prometaphase. RASSF1A interacts with Cdc20, an activator of the anaphase-promoting complex (APC), resulting in the inhibition of APC activity. Although RASSF1A does not contribute to either the Mad2 dependent spindle assembly checkpoint or the function of Emi1 (ref. 1), depletion of RASSF1A by RNA interference accelerated the mitotic cyclin degradation and mitotic progression as a result of premature APC activation. It also caused a cell division defect characterized by centrosome abnormalities and multipolar spindles. These findings implicate RASSF1A in the regulation of both APC-Cdc20 activity and mitotic progression. PMID- 14743219 TI - The DNA damage checkpoint and PKA pathways converge on APC substrates and Cdc20 to regulate mitotic progression. AB - The conserved checkpoint kinases Chk1 and Rad53-Dun1 block the metaphase to anaphase transition by the phosphorylation and stabilization of securin, and block the mitotic exit network regulated by the Bfa1-Bub2 complex. However, both chk1 and rad53 mutants are able to exit from mitosis and initiate a new cell cycle, suggesting that both pathways have supporting functions in restraining anaphase and in blocking the inactivation of mitotic cyclin-Cdk1 complexes. Here we find that the cyclic-AMP-dependent protein kinase (PKA) pathway supports Chk1 in the regulation of mitosis by targeting the mitotic inducer Cdc20. Cdc20 is phosphorylated on PKA consensus sites after DNA damage, and this phosphorylation requires the Atr orthologue Mec1 and the PKA catalytic subunits Tpk1 and Tpk2. We show that the inactivation of PKA or expression of phosphorylation-defective Cdc20 proteins accelerates securin and Clb2 destruction in chk1 mutants and is sufficient to remove most of the DNA damage-induced delay. Mutation of the Cdc20 phosphorylation sites permitted the interaction of Cdc20 with Clb2 under conditions that should halt cell cycle progression. These data show that PKA pathways regulate mitotic progression through Cdc20 and support the DNA damage checkpoint pathways in regulating the destruction of Clb2 and securin. PMID- 14743220 TI - Gadd45 beta mediates the NF-kappa B suppression of JNK signalling by targeting MKK7/JNKK2. AB - NF-kappa B/Rel transcription factors control apoptosis, also known as programmed cell death. This control is crucial for oncogenesis, cancer chemo-resistance and for antagonizing tumour necrosis factor alpha (TNFalpha)-induced killing. With regard to TNFalpha, the anti-apoptotic activity of NF-kappa B involves suppression of the c-Jun N-terminal kinase (JNK) cascade. Using an unbiased screen, we have previously identified Gadd45 beta/Myd118, a member of the Gadd45 family of inducible factors, as a pivotal mediator of this suppressive activity of NF-kappa B. However, the mechanisms by which Gadd45 beta inhibits JNK signalling are not understood. Here, we identify MKK7/JNKK2--a specific and essential activator of JNK--as a target of Gadd45 beta, and in fact, of NF-kappa B itself. Gadd45 beta binds to MKK7 directly and blocks its catalytic activity, thereby providing a molecular link between the NF-kappa B and JNK pathways. Importantly, Gadd45 beta is required to antagonize TNFalpha-induced cytotoxicity, and peptides disrupting the Gadd45 beta/MKK7 interaction hinder the ability of Gadd45 beta, as well as of NF-kappa B, to suppress this cytotoxicity. These findings establish a basis for the NF-kappa B control of JNK activation and identify MKK7 as a potential target for anti-inflammatory and anti-cancer therapy. PMID- 14743222 TI - [Who shall die?]. PMID- 14743221 TI - FAK-Src signalling through paxillin, ERK and MLCK regulates adhesion disassembly. AB - Cell migration is a complex, highly regulated process that involves the continuous formation and disassembly of adhesions (adhesion turnover). Adhesion formation takes place at the leading edge of protrusions, whereas disassembly occurs both at the cell rear and at the base of protrusions. Despite the importance of these processes in migration, the mechanisms that regulate adhesion formation and disassembly remain largely unknown. Here we develop quantitative assays to measure the rate of incorporation of molecules into adhesions and the departure of these proteins from adhesions. Using these assays, we show that kinases and adaptor molecules, including focal adhesion kinase (FAK), Src, p130CAS, paxillin, extracellular signal-regulated kinase (ERK) and myosin light chain kinase (MLCK) are critical for adhesion turnover at the cell front, a process central to migration. PMID- 14743223 TI - [Adults with cerebral palsy--what with them?]. PMID- 14743224 TI - [Can doping be harmful for the heart?]. PMID- 14743225 TI - [Orthopaedic problems in adults with cerebral palsy]. AB - BACKGROUND: There is little information about the need for orthopaedic surgery in adults with cerebral palsy; we wanted to assess this problem. MATERIAL AND METHODS: The material included 37 adults with cerebral palsy (19 women and 18 men), mean age 39. The degree of physical disability was mild in 19 cases, moderate or severe in 18. The patients were assessed by clinical examination and radiographs of hips and spine. RESULTS: 15 patients had pain located to the spine or lower extremities but no-one had severe or invalidating pain. Subjects with moderate or severe disability had a markedly reduced range of motion in the hip and knee joints. Subluxation or dislocation of the hip joints was found in six patients but only one of them had hip pain. Orthopaedic surgery had been performed in 23 patients. In 8 patients we found indications for additional orthopaedic surgery aimed at relieving pain, reducing contractures or improving function. The most frequent procedures would be tenotomies in the hip and knee regions, heel cord lengthening, and triple arthrodesis of the foot. INTERPRETATION: Regular follow-up of adults with cerebral palsy is recommended in order to reveal musculoskeletal problems that can be improved by orthopaedic surgery. PMID- 14743226 TI - [Pyridoxine-dependent seizures]. AB - BACKGROUND: Pyridoxine-dependent seizures is an autosomal, recessively inherited inborn error of metabolism with recurrent long-lasting seizures with onset usually in infancy, but also up to three years of age. The seizures are resistant to conventional anticonvulsants. The condition ends fatally if diagnosis and administration of pyridoxine (vitamin B6 ) in pharmacological doses is delayed too long. MATERIAL AND METHODS: A ten-year-old girl who we believe is affected with this condition is presented. A review of the condition based on relevant literature is given. RESULTS AND INTERPRETATION: The disorder is rare, but may be underdiagnosed. This case report highlights the serious convulsive condition, the problems in diagnosis and treatment, the delayed development before diagnosis and a very positive development during pyridoxine treatment. The condition is variable in clinical expression, and a variety of clinical seizure types may be seen. Diagnosis is clinical and based on response to pyridoxine administration. The pyridoxine dose needed varies, and the aim of treatment is seizure control as well as optimal intellectual development. Prognosis is variable; many are retarded, especially in their speech development. A gene on chromosome 5 is linked to the disorder, but the gene and its product are unknown. PMID- 14743227 TI - [Statin therapy for hypertensive patients]. AB - BACKGROUND: The majority of cardiovascular events and deaths attributable to both raised blood pressure and dyslipidaemia occur in subjects with relatively "normal" blood pressure and lipid levels respectively. The study was designed to evaluate the potential benefits of cholesterol lowering in the primary prevention of coronary heart disease in hypertensive subjects with average and below average levels of serum cholesterol. MATERIAL AND METHODS: Out of 19 342 hypertensive subjects (aged 40-79) who were initially randomized to one of two antihypertensive treatment strategies in the Anglo-Scandinavian Cardiac Outcomes Trial (ASCOT), 10 305 subjects with total cholesterol levels < or = 6.5 mmol/L were additionally randomized to either 10 mg atorvastatin or placebo. The primary endpoint was non-fatal myocardial infarction or fatal coronary heart disease. RESULTS: The lipid arm of the study was prematurely stopped after a median follow up period of 3.3 years. One hundred events occurred in those randomized to atorvastatin compared to 154 events in those receiving placebo, a 36 % relative risk reduction (p = 0.0005) in the primary endpoint. Among secondary and tertiary endpoints, stroke was reduced with 27% (p = 0.02). There was a non-significant 13% reduction in total mortality. Non-cardiovascular mortality was similar in the two treatment limbs of the trial. After three years of follow-up, atorvastatin lowered total serum cholesterol by 1.1 mmol/L compared with placebo. INTERPRETATION: Treatment with 10 mg atorvastatin o.d. in hypertensive patients at moderate risk gives a significant risk reduction of coronary heart disease, independent of baseline level of total cholesterol. PMID- 14743228 TI - [Treatment with statins of the elderly]. AB - BACKGROUND: Previous studies of treatment with statins have included few subjects aged 70 years or above. While the absolute risk of cardiovascular disease in the elderly is very high, the benefits of treatment may be reduced by adverse events, polypharmacy and competing risks. MATERIAL AND METHODS: A statistician and a clinician reviewed the Pravastatin in elderly individuals at risk of vascular disease (PROSPER) study and compared the results with subgroup analyses of previous studies. RESULTS: Subgroup analyses of previous studies showed that treatment with statins reduces cardiovascular events among patients with coronary heart disease aged > or = 65 years. The Heart Protection Study (HPS) included elderly with known atherosclerotic disease, while only 44% of subjects in the PROSPER study had such disease. Among subjects aged 70 or above the difference in events between the groups that received a statin or placebo was 6.1% in the HPS study and 2.1% in the PROSPER study (numbers needed to treat were 6 and 48, respectively). The studies gave conflicting results with regard to stroke and cancer. INTERPRETATION: Elderly people with cardiovascular disease may benefit from treatment with statins. We do not have data that show that statins reduce total mortality among the elderly. PMID- 14743229 TI - [Acute myocardial infarction in a young man who had been using androgenic anabolic steroids]. AB - BACKGROUND: A few case reports suggest that the use of androgenic anabolic steroids may be associated with myocardial infarction. MATERIAL AND METHODS: Case report. RESULTS: We report the case of a 27-year-old male body builder with acute myocardial infarction due to occlusion of the proximal left anterior descending coronary artery. He was treated with primary angioplasty with stent implantation and intra-aortic balloon support, but still developed a large myocardial infarction as determined by both echocardiography and myocardial perfusion tomography. The patient had been using androgenic anabolic steroids regularly for ten years. There was no family history of heart disease or lipid disorder. INTERPRETATION: The actual frequency of myocardial infarction and even sudden death among users of anabolic steroids is presumably underreported in the medical literature. A causal relationship is not established, but a pathogenic role is plausible. Use of androgenic anabolic steroids has been associated with platelet hyperactivity, effects on vasoreactivity and changes in lipid levels. It is important for clinicians to be aware of this association and to counsel patients carefully about this and other side effects that may occur with these agents. PMID- 14743230 TI - [Treatment with percutaneous transluminal angioplasty of the popliteal and crural arteries]. AB - BACKGROUND: The role and benefit of percutaneous transluminal angioplasty (PTA) for treatment of arteriosclerotic lesions in the crural arteries has been controversial. The purpose of this study was to review our experience. MATERIAL AND METHODS: 88 patients were treated with PTA for stenoses and/or occlusions of the popliteal or crural arteries. The indications were disabling claudication in 23, acute ischaemia in 6, and chronic critical ischaemia in 59. Mean age was 75 (46-99), with 32 patients above 80 years of age. The majority had PTA of multiple lesions. Crural arteries were treated in 54 patients (61 %). RESULTS: Technical success was obtained in 111 out of 127 procedures (87%). In patients with claudication there was a clinical success rate of 90%. In chronic critical ischaemia, 30 of 43 treated extremities were preserved after 12 months (70%). Major complications were seen in 3 out of 127 procedures (2.4%). INTERPRETATION: Technically successful PTA is possible in the majority of patients, with few complications. Clinical success can be achieved in patients with high age and advanced arteriosclerosis. Patient selection and cooperation between the vascular surgeon and the radiologist is crucial. : PMID- 14743231 TI - [Ultrasound examination of the shoulder--a method description]. AB - Ultrasound examination of the shoulder in orthopaedics is not much used in Norway, but it is an established method in many institutions abroad. According to the literature, good results have been obtained with diagnostic ultrasound for demonstration of rotator cuff ruptures and abnormalities of the long tendon of the biceps muscle. In order to increase the knowledge of this diagnostic modality in Norway, we present the general principles and techniques of shoulder ultrasonography. Examples of normal and pathological ultrasound findings in the rotator cuff and long tendon of the biceps muscle are given. PMID- 14743232 TI - [Metabolic and cardiovascular adverse effects of modern antipsychotic agents]. PMID- 14743233 TI - [Retinoblastoma--hereditary eye cancer in children]. AB - BACKGROUND: Retinoblastoma is a malignant tumour of the retina that occurs in early childhood. The aim of this paper is to give an updated review of the disease. MATERIAL AND METHODS: A review is given based on literature published over the last few years and on the authors' own experience. RESULTS: The yearly incidence of retinoblastoma is approximately one per 14 000 live births, which gives four new cases of retinoblastoma per year in Norway. The only known risk factor is heritage. Symptoms of retinoblastoma are strabismus, reduced visual acuity and red eye, but the absolutely most important sign is leukokoria (white pupillary reflex). Important diagnostic tools are ophthalmoscopy, ultrasonography, CT and MRI. The goal of treatment is to destroy all tumour tissue, but not the surrounding tissue. Treatment options are enucleation, chemotherapy, external beam radiation, radioactive isotope plaques, cryotherapy, photocoagulation, or a combination of these depending upon the size and location of the tumour. INTERPRETATION: The overall results in the treatment of retinoblastoma are favourable and have improved over the last few years because of better treatment modalities. The survival rate is approximately 95%. It is important that physicians bear in mind the signs of retinoblastoma and especially the alarming sign of leukokoria and acute strabismus in a child. PMID- 14743234 TI - [Ocular changes in Down syndrome]. AB - BACKGROUND, MATERIAL AND METHODS: Down's syndrome is the most common cause of mental retardation with an incidence of about 1.5/1000 live births. Life expectancy and quality of life have improved substantially for this group over the last decades. The aim of this paper is to give an updated short survey of ocular changes present in Down's syndrome based on current international literature and the clinical experience of the authors. RESULTS AND INTERPRETATION: Ocular problems are common, mostly refractive errors, poor accommodation, strabismus, cataract, and keratoconus. Accommodation deficit is present in a majority of individuals with Down's syndrome, also in children and young people. Bifocal or progressive glasses should therefore be prescribed liberally. Because of the high frequency of ocular pathology, all individuals with Down's syndrome should be enrolled in a continuous visual screening programme from birth. We suggest the following screening guidelines: first examination at one month of age, then at one year of age, at 2-3 years of age, at 5-6 years of age (school start), and thereafter every five years. In case of positive findings (e.g. refractive errors, poor accommodation, strabismus) the frequency of examination should be increased and determined individually by the responsible ophthalmologist. PMID- 14743235 TI - [Acute visual loss in an eight-year-old girl]. AB - BACKGROUND: There are many causes of acute visual loss in childhood. Inflammation of the optic nerve, optic neuritis, is rare. While mostly unilateral in adults, the disease often presents bilaterally in childhood. Furthermore, progression to demyelinating disease is unusual and optic neuritis has a better prognosis in childhood than in adulthood. MATERIAL AND METHODS: We describe an 8-year-old girl who over a few days developed painful eyes and profound visual loss. She had afferent pupil defect; ophthalmoscopy showed bilateral papilloedema, characteristic of bilateral optic neuritis. Current literature and guidelines are reviewed with regards to diagnosis and treatment of optic neuritis in childhood. RESULTS: Steroids seem to be beneficial in the treatment of optic neuritis in childhood. Following five days intravenous administration of methylprednisolone and an oral taper over five weeks, visual acuity was almost normalised in both eyes. INTERPRETATION: After studying the literature, we started a treatment with good outcome. Although the prognosis is usually good in children even without treatment, it is important to be aware that there are cases in which treatment with steroids is recommended. PMID- 14743237 TI - [Color blind physicians--is it a problem?]. PMID- 14743236 TI - [Medical certificates issued to help the patient]. AB - BACKGROUND: Medical certificates form the basis for important decisions in society, e.g. the issuing of a driver's licence or the granting of a disability pension. We wanted to know how often doctors deliberately write favourable certificates as seen from the patient's point of view, and why doctors do so. MATERIAL AND METHODS: Questionnaire survey to a representative sample of 1605 Norwegian doctors. Doctors who answered yes to the question above gave information about the frequency of such writing of various types of certificates and their reasons for doing so. RESULTS: 1175 (73%) doctors responded. 39% confirmed deliberately writing in favour of their patients. For most types of certificates, a frequency of less than once a year was most common. 55% admitted writing favourably in disability pension certificates more than once a year, 69% so in referrals. The most common reasons given were securing just treatment and quick access to treatment. Many doctors emphasised that by answering yes, they meant that they were indeed writing expert judgments based on correct facts. INTERPRETATION: Medical certificates deliberately written in favour of the patient is not uncommon. This should lead to a critical discussion of the need for such certificates and medical diagnoses as a basis for resource allocation, rather than the tightening of control over the practice of doctors. PMID- 14743238 TI - [Mercury, fish, fish oil and the risk of cardiovascular disease]. AB - BACKGROUND: Several clinical studies have documented that intake of fish may reduce mortality from coronary heart disease, and two epidemiological investigations have shown a 50% reduction in the incidence of sudden death and of "primary cardiac arrest" in subjects eating fish. However, in some studies no beneficial effects of fish intake on coronary heart disease could be found; one Finnish study even found a positive correlation between intake of freshwater fish and coronary heart disease. One possible explanation for this paradox could be a high content of mercury in fish. MATERIAL AND METHODS: We have studied the relevant literature describing beneficial, less beneficial and negative effects of fish intake on the development of coronary heart disease. Furthermore, we have studied reports that mercury may have properties that enhance the development of coronary heart disease. RESULTS AND INTERPRETATION: Several studies have shown an inverse correlation between omega-3 fatty acids from fish in serum/adipose tissue and coronary heart disease. However, a high content of mercury in hair/toe nail had a negative effect, and in one study the odds ratio for myocardial infarction in those with the highest content of mercury was 2.16. A positive correlation between mercury in hair and the progression of carotid atherosclerosis has been found. Intake of fish is a major source of exposure to mercury, and a high content of mercury probably inhibits the beneficial effects of omega-3 fatty acids on the development of coronary artery disease. PMID- 14743239 TI - [A unified or a varied understanding of disease?]. PMID- 14743240 TI - [Systemic Mycoplasma blood infection in fibromyalgia and chronic fatigue syndrome]. PMID- 14743242 TI - [Cataract extraction--small incisions result in improved vision]. PMID- 14743243 TI - [Blindness in children]. PMID- 14743244 TI - [Primary health care for peace?]. PMID- 14743245 TI - [When not to inform?]. PMID- 14743246 TI - [Guidelines in the clinical work]. PMID- 14743247 TI - [Internship for Norwegian students within the EU/EEC]. PMID- 14743248 TI - [Sexual child abuse]. PMID- 14743250 TI - [Residents in nursing homes and old age homes]. PMID- 14743254 TI - Cytokines, antibodies to cytokines and autoimmunity. AB - It is commonly accepted that cytokines play a major role in the generation of immune responses to various stimuli, whether to exogenous antigens or to autoimmune events. This process must be regulated via complex mechanisms that maintain the normal immune homeostasis. Anti-cytokine antibodies (ACAs) are one of the components that take part in confining immune responses. Manipulating autoimmunity by the administration of ACAs might provide an opportunity to correct an altered regulatory mechanism and to maintain health. The current review will focus on the different ways these objectives might be achieved. PMID- 14743255 TI - Effects of alcohol and other drugs in women of reproductive age: hormonal interactions. AB - Neuroendocrine control of the menstrual cycle is very complex, and the pulsatile hormonal secretory patterns characteristic of the follicular, ovulatory and luteal phases can interact, under certain conditions, with alcohol and other drugs to modulate their effects. The presence of the premenstrual syndrome, with its characteristic symptoms of depression and anxiety, appears to be a critical factor in determining whether alcohol and drug use patterns increase during the premenstruum. PMID- 14743256 TI - Nefazodone and depression. AB - Nefazodone is a fourth-generation antidepressant with novel serotonergic actions. It is a potent antagonist of synaptic 5-HT(2) receptors and a moderate blocker of synaptic 5-HT reuptake. Its mild synaptic blockade of norepinephrine is of little clinical importance. Besides its effectiveness in depression, nefazodone is effective in depression-related anxiety. Of all the antidepressants, nefazodone is least likely to induce sexual dysfunction. This drug increases sleep efficiency and reduces the number of nighttime awakenings. Nefazodone, compared with many other antidepressants, is unlikely to induce a withdrawal syndrome if abruptly stopped. PMID- 14743257 TI - Pulmonary hypertension. AB - Pulmonary hypertension can be caused by an increase in pulmonary blood flow, an increase in left-heart impedance or an increase in pulmonary vascular resistance. Although there are many specific causes that have been identified, clinical presentation and natural history are common to all etiologies. Therapeutic options remain limited to correcting underlying causes whenever possible, as well as supplemental oxygen and, in the setting of severely reduced cardiac output, anticoagulation. Vasodilatory therapy with calcium channel blockers, intravenous epoprostenol or inhaled nitric oxide has also met with some success in patients with primary pulmonary hypertension. Lung or heart-lung transplantation remains the only viable therapeutic option for patients with severe pulmonary hypertension. There have been significant advances in our understanding of the pathobiology and treatment of this uncommon but serious class of disorders. Yet, much remains to be learned in our efforts to improve the lot of these unfortunate patients. PMID- 14743258 TI - Rejection in organ transplantation. AB - Kidney transplantation has been established as a suitable alternative therapy to hemodialysis. Several new immunosuppressive agents contributed to the remarkable improvements in short-term graft survival. However, the rate of graft attrition over the long term has remained constant despite improvements in early success of transplantation during recent years. The half-life of cadaver kidneys, for instance, has consistently remained at 6-7 years throughout the entire transplant experience. Although chronic rejection is the major cause of organ graft loss over the long term, its etiology is not well defined and the pathophysiology of the process is not well understood. Isografts showing similar features of chronic rejection including functional deterioration and morphological changes, demonstrate that antigen-independent factors may also be important in the development of chronic rejection, as well as antigen-dependent factors. PMID- 14743259 TI - The treatment of premature ejaculation: psychological and biological strategies. AB - This review provides criteria for defining premature ejaculation (PE) and identifying men with this dysfunction. Included are discussions of possible PE subtypes and causes of PE. Furthermore, two empirically supported cognitive behavioral approaches for treating PE are described and salient issues surrounding these types of treatment are considered. Biologically based treatments available to PE men are presented, with attention to the relative effectiveness and limitations of the various alternatives. Finally, a model illustrating factors relevant to determining the best therapeutic strategy is described, concluding with a brief discussion of the value of combining biological and psychological modes of treatment. PMID- 14743260 TI - Carvedilol: an effective antihypertensive drug with antiischemic/antioxidant cardioprotective properties. AB - Carvedilol is a multiple-action drug with alpha(1)- and beta-adrenergic receptor antagonistic, calcium channel blocker and antioxidant activities especially useful in the treatment of hypertension. The antioxidant activity of the drug depends on the carbazole moiety, which is unique to carvedilol and is also present in some of the metabolites lacking either the alpha(1)- or beta adrenergic receptor blocking activity. These antioxidant properties account for the exceptional ability of carvedilol to reduce cardiac necrosis in animal models of myocardial infarction, as well as its neuroprotective activities in in vivo and in vitro models of brain ischemia. A further property of carvedilol is its ability to inhibit vascular smooth muscle cell migration and proliferation, and neointima formation, after vascular injuries. The identification of all these activities of carvedilol indicates that it is a singular multiple-action antihypertensive agent with potential for cardiovascular organ protection beyond the normalization of high blood pressure. PMID- 14743261 TI - Estrogen replacement therapy and Alzheimer's disease. AB - Estrogen replacement therapy (ERT) is increasingly recommended for postmenopausal women due to its numerous beneficial effects on bone, cardiovascular system, brain function and quality of life. Data from retrospective epidemiological studies have shown that ERT has a potential to reduce the risk for developing Alzheimer's disease (AD) and to delay its progression. In addition, recent clinical studies have reported improvement of cognitive functions in women with AD. Findings from basic science indicated that the possible mechanisms of action by which estrogen may affect AD include interaction with cholinergic neurotransmitter system, cholinergic neurotrophic and neuroprotective effect, improvement of learning and memory, improvement of cerebral blood flow and metabolism, antioxidant and antiinflammatory action, and interference with beta amyloid protein metabolism and toxicity. Estrogen use in postmenopausal women may offer a new approach for improving cognitive functions in nondemented and demented women, delaying the onset and progression of AD and reducing its occurrence. However, prospective clinical trials are required to establish the efficacy of ERT for prevention and treatment of AD. PMID- 14743262 TI - Inflammatory bowel disease (ulcerative colitis and Crohn's disease): diagnostic criteria and differential diagnosis. AB - Chronic inflammatory bowel diseases (i.e., ulcerative colitis and Crohn's disease) are syndromes in which standardized criteria are necessary in the diagnostic process. The present review is based on the diagnostic criteria used at our institution. We base the diagnosis of ulcerative colitis and Crohn's disease on combined information from the patient history, and radiological, endoscopic and histological findings after exclusion of neoplastic and infectious disease. The patient history must include precise information on the nature and duration of symptoms as well as the presence of relevant influential factors such as travel activity, drug intake and sexual habits. In immunocompromised patients extensive microbiological investigations are required to exclude infection. Typical radiological and colonoscopic findings in ulcerative colitis are mucosal inflammatory changes extending circumferentially and continuously from the rectum and proximally in the colon. In contrast, Crohn's disease is most frequently located in the small bowel and in case of colonic involvement, the rectum is often spared. The best predictors of Crohn's disease are discontinuous lesions, cobblestones and apthous ulceration. Histological changes such as abnormal mucosal architecture and lamina propria cellularity, neutrophil polymorph infiltration and epithelial cell abnormality are useful and reproducible features in the evaluation of colorectal biopsy specimens. The inflammatory bowel diseases, ulcerative colitis and Crohn's disease, continue to be etiological and diagnostic challenges. Increased use of standardized criteria and diagnostic algorithms are essential instruments to improve the overall quality of the management of patients with these diseases. PMID- 14743263 TI - Mode of action and adverse effects of lipid lowering drugs. AB - Serum lipids, cholesterol and triglycerides are incorporated into hydrophilic lipoproteins, which include chylomicrons, very low density lipoproteins (VLDL), intermediate density lipoproteins (IDL), low density lipoproteins (LDL) and high density lipoproteins (HDL). An elevated level of these lipoproteins, except for HDL, is the basis of all hyperlipidemias. However, only some of the lipoprotein fractions, particularly LDL and remnant particles, are potential risk factors for atherogenesis and subsequent cardiovascular disease. Several classes of pharmacological agents are currently available to increase the breakdown and reduce the synthesis of LDL and remnant factors. These include nicotinic acid and its analogs, fibric acid derivatives (e.g., clofibrate, gemfibrozil, bezafibrate), bile acid resins (e.g., cholestyramine), HMG-CoA reductase inhibitors (e.g., lovastatin, simvastatin, pravastatin) and probucol. Lipid lowering drugs of different classes have a synergistic effect on lipid metabolism and combination therapy is often used. Lipid lowering drugs are prescribed as long-term preventive therapy in apparently asymptomatic people. Several studies indicate that secondary prevention with lipid lowering drugs is cost-effective, particularly in patients with symptomatic coronary artery disease. PMID- 14743264 TI - Ocular allergic disease. AB - Ocular allergy is a common condition that usually affects the conjunctiva of the eye and is therefore often referred to as allergic conjunctivitis. The severity of the disease can range from mild itching and redness, as seen in seasonal allergic conjunctivitis, to the more serious vision threatening forms of ocular allergy which affect the cornea, such as atopic keratoconjunctivitis. The pathogenesis of allergic conjunctivitis involves a complex mechanism which centers around IgE-mediated mast cell degranulation and release of multiple preformed and newly formed inflammatory mediators. The diagnosis of allergic conjunctivitis is usually a clinical one which can be made based on a thorough history and careful examination. Treatment of ocular allergy should begin with conservative measures including allergen avoidance, environmental control, ocular irrigation and cold compresses. Pharmacotherapy of allergic conjunctivitis consists of several classes of drugs. Antihistamines are widely used to treat mild conditions such as seasonal and perennial conjunctivitis and potent new agents such as levocabastine and emedastine are now available. Mast cell stabilizers such as sodium cromoglycate are both safe and effective and are commonly used in ocular allergy. More effective mast cell stabilizers such as nedocromil, lodoxamide and olopatadine are now being used. Nonsteroidal antiinflammatory drugs have demonstrated only limited efficacy and, as such, are not widely used. Topical steroids are very effective in treating signs and symptoms but are reserved for only refractory cases due to their serious side effects. Loteprednol and rimexelone are newer corticosteroids which reportedly have less of an effect on intraocular pressure. Cyclosporine has recently been shown to be highly effective in cases of vernal keratoconjunctivitis and atopic keratoconjunctivitis while producing no adverse effects. PMID- 14743265 TI - Pharmacological and clinical profile of valsartan. AB - The development of angiotensin II (Ang II) receptor antagonists is one of the latest advances in the pharmacological treatment of hypertension and other related diseases. Since the antagonism of angiotensin biological activity has been shown to represent a potent way to favorably affect systemic hypertension, its use is becoming more and more widespread among clinical practice. Besides the originally developed inhibitors of angiotensin II converting enzyme, today molecules that are effective in selectively blocking the type I angiotensin II receptor are available. In this report we describe the main characteristics of one of these compounds, the angiotensin receptor antagonist valsartan, in terms of pharmacological profile as well as efficacy and tolerability. The main purpose is to provide a comprehensive description of the knowledge gained during several years of experimental and clinical studies that can be useful for guidance during the choice for treatment of hypertension, as well as to obtain other potential beneficial effects of this drug on progressive organ damage related to hypertension. PMID- 14743266 TI - Advances in hypertension management: update 1998. AB - This article briefly reviews some important issues in the management of hypertension recently published or presented in medical congresses. The main results of several large outcome clinical trials, such as the Systolic Hypertension in Europe (Syst-Eur) trial, Hypertension Optimal Treatment (HOT) study and the Captopril Prevention Project (CAPPP) study, are summarized. The results of several retrospective studies dealing with the risk of cancer associated with antihypertensive treatments are presented. Finally, the nephroprotective effects of angiotensin-converting enzyme inhibitors in the progression of nondiabetic renal disease and the controversy surrounding the cardiovascular risk associated with dihydropyridine calcium antagonists are discussed. PMID- 14743267 TI - Disseminated intravascular coagulation: diagnosis, treatment and related pathophysiology. AB - This review presents a current evaluation of disseminated intravascular coagulopathy from approaches involving clinical and differential diagnosis, pharmacological treatment, biological markers, disease effects on various organs, risk factors and related pathophysiological conditions. While the optimal regimens for treatment with blood components and the absolute indications for anticoagulant and antifibrinolytic treatments are unknown, various studies are discussed which offer promising new drugs for the present and the future. Associations between this disease and other pathophysiologies are presented which provoke new perspective and insight into disease occurrence and progression. PMID- 14743268 TI - The chlamydial pneumonias. AB - Chlamydia psittaci and Chlamydia pneumoniae are important causes of community acquired pneumonias. Less commonly, C. trachomatis may cause pneumonia in adult immunocompromised hosts but more commonly is responsible for pneumonia in neonates. C. psittaci is the cause of psittacosis and is the only chlamydial zoonotic atypical pneumonia. C. pneumoniae is being increasingly recognized as the cause of up to 10% of community-acquired pneumonias. C. pneumoniae pneumonia has a clinical presentation like Mycoplasma pneumoniae pneumonia. C. pneumoniae is also responsible for a variety of other respiratory tract infections, e.g., sinusitis, bronchitis, otitis, pharyngitis and laryngitis. C. pneumoniae, like M. pneumoniae, may result in permanent airway disease, e.g., asthma, following infection. All chlamydia are sensitive to doxycycline. Macrolides are highly active against C. trachomatis, and in spite of in vitro susceptibility, are relatively inactive in vivo against C. psittaci and C. pneumoniae. Fluoroquinolones are also active against chlamydia. Doxycycline remains the preferred antibiotic to treat all chlamydial infections in nonpregnant adults. PMID- 14743269 TI - Topical antiviral agents for herpes simplex virus infections. AB - Several antiviral agents against herpes simplex virus (HSV) infection have been clinically studied. Earlier therapies include gluteraldehyde, povidone-iodine, butylated hydroxytoluene and ether. Nucleoside analogs have been tested for efficacy in HSV. Although acyclovir and adenine arabinoside have shown minimal therapeutic benefit, cidofovir has been successful in the treatment of acyclovir resistant strains of HSV, and idoxuridine 15% in dimethyl sulfoxide, edoxudine and penciclovir have significant clinical benefit against HSV. Interferon-alpha has shown synergism with other anti-HSV drugs such as caffeine, trifluorothymidine, dimethyl sulfoxide and nonoxynol-9, and ascorbic acid shows promising effects against HSV. Using a vehicle that enhances skin penetration of a drug or further exploring combination therapy may result in efficacious treatment of HSV. Vaccination or gene therapy may also prove beneficial in future studies. PMID- 14743270 TI - Zolmitriptan clinical studies. AB - Zolmitriptan, a selective 5-HT(1B/D) agonist was developed for the acute treatment of migraine. Dose-finding studies show a clearly defined dose response curve for the oral formulation with onset of efficacy demonstrated within 45 min of dosing. Clinical trials support its efficacy in all types of migraine, with excellent safety and tolerability in those patients for whom zolmitriptan is not contraindicated. Future developments, including new formulations, will provide patients with a greater choice of treatment. PMID- 14743271 TI - Environmental effects of ozone depletion and its interactions with climate change: progress report 2003. AB - United Nations Environment Programme, Environmental Effects Assessment Panel PMID- 14743272 TI - Photochemical alpha-cleavage of ketones: revisiting acetone. AB - The photochemical [small alpha]-cleavage of acetone is analyzed in view of recent results obtained for the isolated molecule in supersonic jets. The fluorescence decay time of the isolated molecule spans a range of more than six orders of magnitude, from approximately 10(-6) s near the origin of the S(0)-S(1) transition to less than 10(-12) s at about 20 kcal x mol(-1) excess energy. In contrast, the decay time of the excited singlet (S(1), (1)n pi) in the bulk is around 10(-9) s and independent of excitation wavelength. Initial excitation to the (1)npi state is followed by internal conversion (IC) to the ground state and intersystem crossing to the lowest-lying triplet. The rate constants of these processes are comparable to the radiative decay rate constant for excess energy up to 7 kcal x mol(-1) above the origin of the S(0)-S(1) transition. Beyond that energy, the triplet state becomes dissociative and the ISC rate becomes much larger than other processes depleting S(1). The primary reaction on the triplet surface is a barrier-controlled alpha-cleavage to form the triplet radical pair CH(3)(*)+ CH(3)CO(*). Direct reaction from the S(1) is negligible, and the non quenchable reaction (by triplet quenchers) observed in the bulk gas phase is due to hot triplet molecules that dissociate on the timescale of 10(-12) s or less. The singlet-state decay time measured in the bulk (approximately 1-2 ns) arises from collision-induced processes that populate low-lying levels of S(1). The analysis is aided by detailed state-resolved studies on related molecules (in particular formaldehyde and acetaldehyde) whose photophysics and photochemistry parallel those of acetone. PMID- 14743273 TI - Reactive species formed on proteins exposed to singlet oxygen. AB - Singlet oxygen ((1)O(2)) is believed to be generated in biological systems by a range of endogenous processes (e.g. enzymatic and chemical reactions) and exogenous stimuli (e.g. UV or visible light in the presence of a sensitiser). Kinetic data is consistent with proteins being a major target for (1)O(2), with damage occurring preferentially at Trp, His, Tyr, Met, and Cys side-chains. Reaction with each of these residues gives rise to further reactive species. In the case of Trp and Tyr, initial poorly characterised endoperoxides are believed to undergo ring-opening reactions to give hydroperoxides, which can be reduced to the corresponding alcohols; other products arising from radical intermediates can also be generated, particularly in the presence of UV light and metal ions. With His side-chains, poorly characterised peroxides are also formed. Reaction with Met and Cys has been proposed to occur via zwitterionic peroxy intermediates. Peroxides are also generated on isolated proteins, and protein within intact cells, via(1)O(2)-mediated reactions. The peroxides formed on Trp, Tyr, and His peptides, as well as on proteins, have been shown to induce damage to other targets, with molecular oxidation of thiol residues an important reaction. This can result in the inactivation of cellular enzymes and the oxidation of other biological targets. Protein cross-linking and aggregation can also be induced by reactive species formed on photo-oxidised proteins, though the nature of the species that participate in such reactions is poorly understood. These secondary reactions, and particularly those involving hydroperoxides, may play a key role in the induction of secondary damage (bystander effects) in systems subject to photo-oxidation. PMID- 14743274 TI - Bi-directional photocurrent generation dependent on the wavelength of irradiation of a mixed monolayer assembly. AB - A mixed self-assembled monolayer consisting of a ruthenium tris-bipyridine complex linked to viologen and a palladium phthalocyanine derivative was fabricated on a gold electrode by means of pendant thiol groups. The direction of the photocurrent which is induced in the electrode when it is irradiated depends on the wavelength of the light used. PMID- 14743275 TI - Excimer probe of the binding of alkyl disulfides to gold nanoparticles and subsequent monolayer dynamics. AB - A symmetric alkyl disulfide bearing two pyrenyl chromophores allows for fluorescence monitoring of its binding to gold nanoparticles and the subsequent diffusion of the geminate thiolates on the particle surface. PMID- 14743277 TI - Stereoselectivity in the triplet decay of chiral benzophenone-naphthalene bichromophoric systems. AB - Chiral recognition in the intramolecular induced quenching of the methoxynaphthalene triplet by benzophenone has been observed by using diastereomeric bichromophores PMID- 14743276 TI - Intracellular localization of a long alkyl chain tetraphenylporphyrin and chloride channel activation in Psammobatis extenta electrocytes. AB - The intracellular localization of a tetraphenylporphyrin bearing four long lipophilic alkyl chains in electrocytes from Psammobatis extenta(Rajidae) is described. In contrast to what is usually the case, this porphyrin derivative is localized in the electromotor nerves and the nuclear chromatin of the electrocytes. Both structures exhibited intense fluorescence, whereas, the mitochondria were only slightly fluorescent. These data are discussed in relation to electrocyte death in a weakly electric fish. Additionally, electron probe X ray microanalysis suggests a migration of chloride and cationic ions, which might be implicated in chloride and cationic channel activation in the electrocyte. PMID- 14743278 TI - Biexciton resonance energy transfer in a model photosystem. AB - The absorption of two photons by a concentrated assembly of identical chromophores can lead to formation of a biexciton. This work concerns the transfer of biexcitonic energy from a ring to a central donor, with a particular focus on rates and geometric effects. Quantum amplitudes are expressed in terms of orientation factors with a clear physical significance and detailed calculations are performed on an idealised, three-fold symmetric photosystem-a structural motif common in a variety of natural and artificial light-harvesting systems. PMID- 14743279 TI - Multiphoton-excited luminescence of a lanthanide ion in a protein complex: Tb(3+) bound to transferrin. AB - The Tb(iii) complex of the iron-transport protein transferrin (Tb(2)-Tfr) exhibits strongly sensitised, sharp line luminescence from f-f states following multiphoton excitation via two tyrosinate residues directly co-ordinated to the lanthanide ion. Using an ultrafast Ti:sapphire laser system, a quadratic dependence of the Tb(iii) luminescence intensity was observed on excitation with photons at 503 and 566 nm, and a cubic dependence with photons at 800 nm. The two photon cross-sections at 503 and 566 nm are 7.4 x 10(-50) and 0.37 x 10(-50) cm(4) s photon(-1) mol(-1), respectively, which compare favourably with values reported for the green fluorescent protein. Three-photon excitation at 800 nm gives rise to a Tb(iii) emission spectrum with excellent signal to noise ratios. These results lead to a proposal that if a Tb(iii)-protein complex with similar luminescent properties could be formed in vivo, an intra-cellular imaging system that uses multiphoton-excited, long-lived lanthanide ion luminescence could be developed. This offers the prospect of multiphoton imaging in tight focal planes using sharp line emission with long lifetimes for wavelength and time discrimination against background fluorescence. PMID- 14743280 TI - Photoelectrochemical properties of supramolecular species containing porphyrin and ruthenium complexes on TiO(2) films. AB - Modification of wide band gap semiconductor surfaces by a new generation of supramolecular sensitizers, combining porphyrin and ruthenium-phenanthroline complexes leads to versatile molecular interfaces, allowing the exploitation of photoinduced charge transfer in dye sensitized photoelectrochemical cells. meso Tetrapyridylporphyrin coordinated to two ruthenium complexes converts 21% of the incident photons into current after excitation at the Soret band. In this work we discuss the electron/energy transfer mechanisms involved in the TiO(2) sensitization by these supramolecular species, invoking some theoretical calculations. PMID- 14743281 TI - Bimolecular processes on silica gel surfaces: energetic factors in determining electron-transfer rates. AB - Triplet state and radical cation formation is observed following laser excitation of anthracene, phenanthrene and naphthalene (and their derivatives) adsorbed on silica gel. Energy- and electron-transfer reactions of these compounds with co adsorbed azulene have been studied using a time-resolved diffuse reflectance laser flash photolysis technique. Triplet energy transfer from the arene derivative to azulene and electron transfer from azulene to the arene radical cation have been investigated in order to distinguish between diffusional and energetic control in these systems. Energy and electron transfer can be studied independently due to differing absorption properties and energy dependencies of production of the triplet states and radical cations. Transient decay kinetics for both electron and energy transfer have been modelled using two different rate constant distributions: a log Gaussian and a symmetrical Levy stable distribution. The latter model has also been demonstrated to be applicable to the decay of radical cations in the absence of an electron donor, which cannot be adequately described by the Gaussian model. Energy-transfer rates between the arene derivatives and azulene have been found to be close to the diffusion controlled limit; however, in most cases, the rate of electron transfer is considerably lower. A correlation between the bimolecular rate constant and free energy of electron transfer has been found, indicating a Marcus inverted region. Compounds with bulky substituents show a further reduction in the rate of electron transfer, suggesting that an additional steric factor is involved in this process. PMID- 14743282 TI - Photoprocesses of p-naphthoquinones and vitamin K(1): effects of alcohols and amines on the reactivity in solution. AB - The photochemistry of 1,4-naphthoquinone (NQ), the 2-methyl, 2,3-dichloro and 2 bromo derivatives, and vitamin K(1) was studied in non-aqueous solvents by time resolved UV-vis spectroscopy after ns laser pulses at 248 and 308 nm. The triplet state of the NQs reacts with alcohols and amines, e.g. triethylamine (TEA) and DABCO, yielding semiquinone radicals (HQ(*)/Q (*)(-)). They are the major intermediates and their second-order decay kinetics depend on the properties of the additives and the medium. Transient conductivity measurements suggest the occurrence of photoinduced electron transfer from amines to the triplet state of NQs in acetonitrile. The photoconversion lambda (irr)= 254 nm) of NQs to the 1,4 dihydroxynaphthalenes (H(2)Q) was measured in the absence and presence of varying concentrations of electron and H-atom donors, and the quantum yield was found to increase with increasing electron- or proton-donor concentration. The mechanisms of photoreduction of NQs by propan-2-ol and TEA in acetonitrile exhibit a number of similarities. Oxygen quenches the triplet state, thereby forming singlet molecular oxygen. Oxygen also reacts with the semiquinone radical, thereby forming HO(2)(*)/O(2) (*) (-) radicals, and reacts with H(2)Q, thereby re-forming the quinone. A different pattern, involving intramolecular H-atom transfer, holds for vitamin K(1), where 1,3-quinone methide (1,3-QM) diradicals were observed in acetonitrile prior to formation of two 1,2-QM tautomers, but a triplet was not. The decay of the 1,3-QM intermediates becomes faster in the presence of alcohols and amines due to proton-transfer reactions. PMID- 14743283 TI - Solvent effects on the photophysics of 3-(benzoxazol-2-yl)-7-(N,N diethylamino)chromen-2-one. AB - The photophysics of 3-(benzoxazol-2-yl)-7-(N,N-diethylamino)chromen-2-one was studied in different solvents and in SDS micelles. This compound presents characteristics which include an S(0)---> S(1) ( pi,pi*) transition with a (1)(n,pi*) perturbative component, due to the electronic coupling between the diethylamino group and the coumarin ring, considerable solvatochromism, dual fluorescence and high fluorescence quantum yields in almost all solvents studied. The electronic structure of the S(1) and S(2) excited states permits vibronic coupling between them, making configurational changes of the S(2) excited state possible, leading to the formation of an S(2)(TICT) state. Analysis of the TCSPC data indicates an equilibrium between the S(2)(TICT) and S(1)(LE) states in favour of the former. In protic solvents, the hydrogen bonding between the solvent and the diethylamino moiety results in the formation of an S(2)(HICT) state, making internal conversion an important deactivation process. Quantum mechanical calculations for the isolated molecule show that the diethylamino group in the S(2)(TICT) state is twisted at least 56 degree from the plane of the coumarin ring, with partial electronic decoupling between -NEt(2) and the coumarin ring. This twisting angle must be positively influenced by solute solvent interactions. [capital Phi](ST) is found to be small, but not negligible. However, Phi (delta) can be considered negligible, an indication that T(1) is a short-lived state. Based on the experimental data and theoretical calculations, the most probable sequence for the first excited states, including the TICT state, is T(1)(n, pi*) < S(2)(TICT) < S(1)(pi,pi*) approximate S(2) (n,pi*). PMID- 14743284 TI - Immunochemical study of DNA modifications in the nuclei of UV-damaged lymphocytes. AB - Studies of UV-induced skin cancers show that malignisation of skin cells, as well as alterations in anti-tumor immune control, are triggered by UV-induced lesions in cellular DNA. Such lesions can probably appear in the human mononuclear leukocytes (lymphocytes) during exposure of skin to sunlight. With the aim of studying the processing of UV-induced DNA lesions in these cells, we used flow cytometry and labelling of their partially denatured nuclei with the monoclonal antibody (H3) that binds cyclobutane pyrimidine dimers in single-stranded DNA. After the first few hours of cultivation of the irradiated cells, we found an increase in H3-specific fluorescence from cellular nuclei, while there was a decrease in the number of H3-positive sites in isolated DNA from these cells. We examined cells cultured under different conditions and concluded that the effect of enhancement of H3 labelling of nuclei did not result from changes in temperature and culture medium. Furthermore, we have found that this effect, as well as the decrease in H3 labelling in isolated DNA, are both prevented by pretreatment of the cells with Novobiocin, which we used as an inhibitor for the topoisomerase II-induced relaxation of supercoiled DNA prior to repair-specific incision. The inhibition by Novobiocin of the above-mentioned changes in H3 labelling in cellular nuclei and isolated DNA of the irradiated cells clearly indicate the association of both effects with an excision repair-related DNA modification. While the partial loss of H3-binding sites from isolated DNA is obviously a result of excision of some fraction of pyrimidine dimers, the enhancement of the H3 labelling of nuclei might be due to the formation of open structures at dipyrimidine-containing DNA fragments in preparation for incision. We suggest that formation of open structures predominates quantitatively over dual incision and excision of these fragments, and leads to enhanced exposure of the pyrimidine dimers in nuclei to H3 binding. Thus, unstimulated human lymphocytes appear to be capable of performing pre-incision steps for removal of these DNA lesions. PMID- 14743285 TI - Exposure to multiple doses of UVB radiation reduces the numbers of epidermal Langerhans cells and lymph node dendritic cells in mice. AB - Immune suppression following UVB irradiation is partly attributed to the effects of the exposure on antigen-presenting cells. Following a single UVB irradiation, there is a decrease in epidermal Langerhans cell numbers; this is accompanied by an increase in the number of dendritic cells (DC) in lymph nodes draining the irradiated site. We investigated whether a similar effect occurred following multiple UVB exposures. Mice were irradiated on their ears and shaved dorsal skin twice a week for 3 weeks. After the final exposure, the number of ATPase(+) Langerhans cells in epidermal sheets prepared from the ears was found to be decreased by 33% compared to unirradiated controls. The number of DC in the draining lymph nodes (DLN) did not increase as might have been expected; rather, a significant decrease of approximately 30% in DC numbers in the DLN of UVB irradiated mice compared with unirradiated controls occurred. This decrease in antigen-presenting cells in both the epidermis and the DLN may be an important contributing factor to the immune suppression that follows multiple UVB exposures. PMID- 14743286 TI - Enhancement of the blood growth promoting activity after exposure of volunteers to visible and infrared polarized light. Part I: stimulation of human keratinocyte proliferation in vitro. AB - The systemic mechanisms of the wound healing effect of low intensity lasers remain largely uninvestigated. The goal of this randomized, placebo controlled, double blind study is to prove that irradiation of a small area of the human body with visible and infrared polarized (VIP) light (400-3400 nm, 95% polarization, 40 mW cm(2), 12 J cm(2)) leads to an increase of the growth promoting (GP) activity of the entire circulating blood for primary cultures of human keratinocytes (KCs). Thirty minutes after the VIP-irradiation of a sacral area of volunteers, the GP activity of circulating blood was seen to increase through the elevation of the number of KCs cultured with the isolated plasma by 20 +/- 3%, p < 0.001. A similar increase in GP activity was seen in plasma derived from the in vitro irradiated blood of each volunteer, and from the mixture of irradiated and non-irradiated autologous blood 1:10. Enhanced GP activity was also recorded at 24 h after the 1st and 4-9th daily phototherapeutic sessions. Hence, exposure of volunteers to VIP light leads to a fast increase in the GP activity of the entire circulating blood for human KCs in vitro, which is a consequence of the transcutaneous photomodification of blood and its effect on the rest of the circulating blood volume. PMID- 14743290 TI - Community capacity for adaptation to climate-induced water shortages: linking institutional complexity and local actors. AB - There is growing concern for the capacity of urban and rural communities to manage current water shortages and to prepare for shortages that may accompany predicted changes in climate. In this paper, concepts relating to the notion of climate adaptation and particularly "capacity building" are used to elucidate several determinants of community-level capacity for water management. These concepts and criteria are then used to interpret empirically derived insights relating to local management of water shortages in Ontario, Canada. General determinants of water-related community capacity relate to upper tier political and institutional arrangements; the characteristics of, and relationships among, pertinent agencies, groups, or individuals involved in water management; and the adequacy of financial, human, information, and technical resources. The case analysis illustrates how general factors play out in local experience. The findings point to geographically specific factors that influence the effectiveness of management. Key factors include collaboration between water managers, clarification of agency roles and responsibilities, integration of water management and land-use planning, and recognition and participation of both urban and rural stakeholders, whose sensitivities to water shortages are spatially and temporally variable. PMID- 14743292 TI - A complex systems model approach to quantified mineral resource appraisal. AB - For federal and state land management agencies, mineral resource appraisal has evolved from value-based to outcome-based procedures wherein the consequences of resource development are compared with those of other management options. Complex systems modeling is proposed as a general framework in which to build models that can evaluate outcomes. Three frequently used methods of mineral resource appraisal (subjective probabilistic estimates, weights of evidence modeling, and fuzzy logic modeling) are discussed to obtain insight into methods of incorporating complexity into mineral resource appraisal models. Fuzzy logic and weights of evidence are most easily utilized in complex systems models. A fundamental product of new appraisals is the production of reusable, accessible databases and methodologies so that appraisals can easily be repeated with new or refined data. The data are representations of complex systems and must be so regarded if all of their information content is to be utilized. The proposed generalized model framework is applicable to mineral assessment and other geoscience problems. We begin with a (fuzzy) cognitive map using (+1,0,-1) values for the links and evaluate the map for various scenarios to obtain a ranking of the importance of various links. Fieldwork and modeling studies identify important links and help identify unanticipated links. Next, the links are given membership functions in accordance with the data. Finally, processes are associated with the links; ideally, the controlling physical and chemical events and equations are found for each link. After calibration and testing, this complex systems model is used for predictions under various scenarios. PMID- 14743291 TI - Understanding fish behavior, distribution, and survival in thermal effluents using fixed telemetry arrays: a case study of smallmouth bass in a discharge canal during winter. AB - Advances in telemetry have facilitated the continuous monitoring of fish position and movement. At present, there are few examples where this approach has been applied to environmental monitoring or assessment. Here we 1) present a case study that used a fixed antenna array and continuously scanning coded receiving system to monitor the movement of radio-tagged smallmouth bass ( Micropterus dolomieu) in a thermal discharge canal on Lake Erie during the winter of 1998/1999, and 2) evaluate the use of fixed telemetry arrays for environmental monitoring. Although the number of radiotagged bass in the canal decreased gradually over time, fish spent the majority of the winter in the canal. When in the canal, bass selected areas upstream of the tempering pumps where water was the warmest. This region was also high in habitat complexity, had adequate velocity refuges, and abundant forage. Despite residing in the thermal effluent throughout the winter, none of the fish monitored were observed to participate in reproductive activities in the canal in the Spring. Interestingly, during a biofouling chlorination pulse in May, 50% of radiotagged fish still residing in the canal left and did not return during the monitoring period. Utility infrastructure accessible to fish, including thermal effluents, should be considered as fish habitat and managed accordingly to minimize mortality and sublethal effects on resident and transient fish. Fixed telemetry arrays that permit the continuous monitoring of fish behavior as described in this paper are widely applicable to many issues in environmental management, monitoring, and conservation. PMID- 14743303 TI - Performance of the chronically hypoxic young rabbit heart. AB - Hearts isolated from 30 rabbits, raised from birth to approximately 5 weeks of age under either hypoxic (FIO2, 0.10) or normoxic (FIO2, 0.21) conditions, underwent retrograde aortic perfusion using a non-recirculating, well-oxygenated crystalloid solution. The left ventricular end diastolic pressure was initially set at approximately 5 mmHg. Aerobic performance was studied by measuring peak systolic pressure (PSP), coronary flow, glucose oxidation, and oxygen consumption. Anaerobic function was assessed by determining time for the onset of contracture (TOC) in the presence of zero coronary flow. Hypoxic vs normoxic hearts (mean+/-SEM): heart rate, 197+/-6 vs 190+/-5 beats per minute; PSP, 136+/ 4* vs 108+/-4 mmHg; dP/dt(max), 2294+/-125* vs 1549+/-144 mmHg/sec; relaxation time constant (Tau), 26.9+/-1.1* vs 41.6+/-4.8 msec; (-) dP/dt(max), 1422+/-43* vs 1001+/-63 mmHg/sec; coronary flow, 86.3+/-4.2* vs 59.9+/-2.9 ml/min/g(dry); glucose oxidation, 3511+/-118* vs 2979+/-233 nmol/min/g(dry); oxygen consumption, 28.2+/-1.4* vs 22.7+/-1.4 micromol/min/g(dry); and TOC, 11.8+/-1.2* vs 22.9+/-2.2 min (*p < 0.05). Hearts isolated from young rabbits, exposed to hypoxia from birth, exhibited enhanced ventricular systolic and diastolic mechanical function, elevated coronary flow, retained capacity for aerobic metabolism, and a shorter TOC compared to their normoxic counterparts. PMID- 14743304 TI - Severe pulmonary regurgitation late after total repair of tetralogy of Fallot: surgical considerations. AB - BACKGROUND: After total repair of tetralogy of Fallot (TOF-R) with transannular patching (TAP), severe pulmonary regurgitation (PR) is reported to develop in up to 30% of patients at a follow-up of 20 years, and 10-15% or more need pulmonary valve replacement (PVR). In this study, time-related progression of PR and right ventricular (RV) dilatation, and functional recovery of the RV after PVR are analyzed, and the possible causes of PR and timing of PVR are discussed. METHODS: Eighteen patients, who late after TOF-R with TAP underwent PVR for severe PR, were chosen for the study. NYHA class, QRS duration, RV dilatation index (RVDI = RVEDD/LVEDD), and RV-distal pulmonary artery (PA) peak systolic gradient were reviewed and retrospectively analyzed. RESULTS: TOF-R was performed at a mean age of 5.1 +/- 3.9 years (range: 0.6-12.8 years); the mean time interval from TOF-R to PR grade 3 onset was 11.8 +/- 7.0 years (range: 3.3-27.4 years), and from TOF R to PVR was 18.5 +/- 7.8 years (range: 8.7-37.1 years). At PVR, 11 patients were in NYHA class II-III, all patients had severe PR (grade 3/3) and severe RV enlargement, 4 patients had ventricular arrhythmias, 7 patients significant distal pulmonary artery stenosis, and 2 patients small nonrelevant residual VSD. The mean preoperative RVDI (normal: 0.5) was 0.99 +/- 0.14 (range: 0.75-1.3), the mean QRS duration 170 +/- 24 ms (140-220 ms), and the mean RV-distal PA peak systolic pressure gradient 33.3 +/- 19.0 mmHg (range: 10-60 mmHg). Patients aged at TOF-R> 5 years had considerably longer redo-free intervals than their younger counterparts: mean 23.1 years (range 8.7-37.1 years) vs 14.8 years (range: 9.3 21.2 years), respectively. The redo-free intervals and the duration of severe PR correlated inversely with the RV-PA gradient. At a mean follow-up of 1.3 years (2 weeks-5 years), the mean RVDI decreased from 0.99 +/- 0.14 to 0.69 +/- 0.15, the mean validity class improved from 2.5 to 1.1. One patient died. CONCLUSIONS: After TOF-R with TAP, the progression of PR has very individual dynamics, resulting in extremely varying redo-free intervals. Concomitant pulmonary stenosis seems to exaggerate progression of PR. PVR results in effective reduction of diastolic dimensions of severely dilated RV and in improvement of validity class. Referred PVR in no-risk cases seems to be justified. PMID- 14743306 TI - Repetitive paroxysms of supraventricular tachyarrhythmias triggered during pediatric cardiac interventions: suppression after short infusion of amiodarone. AB - Arrhythmias are the most common major complications encountered during pediatric cardiac catheterizations. This report describes the management of repetitive paroxysms of supraventricular tachyarrhythmias triggered by catheter manipulation during interventional procedures in two children. After 15 minutes, amiodarone infusion (25 microg/kg/min) eliminated further paroxysms in both patients, allowing both interventions to be completed safely and effectively. PMID- 14743307 TI - Eosinophilic pericarditis: a rare complication of idiopathic hypereosinophilic syndrome in a child. AB - We describe an 18-month-old child with idiopathic hypereosinophilic syndrome (IHES) who presented with fever, and cervical lymphadenopathy. Chest X-ray showed marked cardiomegaly, and echocardiogram revealed large pericardial effusion. Other causes of pericarditis were excluded. Despite the initiation of steroid therapy, signs of impending cardiac tamponade developed. Pericardiocentesis yielded bloody fluid with a white blood count of 14,800/mm3, of which 23% were eosinophils. The child recovered after pericardial drainage and prolonged systemic steroid therapy. Eosinophilic pericarditis is a rare but potentially dangerous complication of IHES. PMID- 14743308 TI - Persistent fifth aortic arch and fourth arch interruption in a 28-year-old woman. AB - A 28-year-old woman having persistent fifth aortic arch and aortic coarctation associated with the fourth arch interruption, without other cardiac malformation, underwent graft interposition, 20 mm in diameter, between the fourth arch including the left subclavian artery and the descending aorta. PMID- 14743309 TI - Cost-effectiveness of Project ADAM: a project to prevent sudden cardiac death in high school students. AB - Public access defibrillation (PAD) in the adult population is thought to be both efficacious and cost-effective. Similar programs aimed at children and adolescents have not been evaluated for their cost-effectiveness. This study evaluates the potential cost-effectiveness of implementing Project ADAM, a program targeting children and adolescents in high schools in the Milwaukee Public School System. Project ADAM provides education about cardiopulmonary resuscitation (CPR) and the warning signs of sudden cardiac death (SCD) and training in the use and placement of automated external defibrillators (AEDs) in high schools. We developed decision analysis models to evaluate the cost effectiveness of the decision to implement Project ADAM in public high schools in Milwaukee. We examined clinical model and public policy applications. Data on costs included estimates of hospital-based charges derived from a pediatric medical center where a series of patients were treated for SCD, educational programming, and the direct costs of one AED and training for 15 personnel per school. We performed sensitivity analyses to assess the variation in outputs with respect to changes to input data. The main outcome measures were Life years saved and incremental cost-effectiveness ratios. At an arbitrary societal willingness to pay $100,000 per life year saved, the policy to implement Project ADAM in schools is a cost-effective strategy at a threshold of approximately 5 patients over 5 years for the clinical model and approximately 8 patients over 5 years for the public policy model. Implementation of Project ADAM in high schools in the United States is potentially associated with an incremental cost-effectiveness ratio that is favorable. PMID- 14743310 TI - Whole proteome prokaryote phylogeny without sequence alignment: a K-string composition approach. AB - A systematic way of inferring evolutionary relatedness of microbial organisms from the oligopeptide content, i.e., frequency of amino acid K-strings in their complete proteomes, is proposed. The new method circumvents the ambiguity of choosing the genes for phylogenetic reconstruction and avoids the necessity of aligning sequences of essentially different length and gene content. The only "parameter" in the method is the length K of the oligopeptides, which serves to tune the "resolution power" of the method. The topology of the trees converges with K increasing. Applied to a total of 109 organisms, including 16 Archaea, 87 Bacteria, and 6 Eukarya, it yields an unrooted tree that agrees with the biologists' "tree of life" based on SSU rRNA comparison in a majority of basic branchings, and especially, in all lower taxa. PMID- 14743311 TI - Modeling DNA base substitution in large genomic regions from two organisms. AB - We studied the substitution patterns in 7661 well-conserved human-mouse alignments corresponding to the intergenic regions of human chromosome 22. Alignments with a high average GC content tend to have a higher human GC content than mouse GC content, indicating a lack of stationarity. Segmenting the alignments into four groups of GC content and fitting the general reversible substitution model (REV) separately gave significantly better fits than the overall fit and the levels of fit are close to that expected under an REV model. In addition, most of the fitted rate matrices are not of the HKY type but are remarkably strand-symmetric, and we constructed a number of substitution matrices that should be useful for genomic DNA sequence alignment. We did not find obvious signs of temporal inhomogeneity in the substitution rates and concluded that the conserved intergenic regions in human chromosome 22 and mouse appear to have evolved from their common ancestors via a process that is approximately reversible and strand-symmetric, assuming site homogeneity and independence. PMID- 14743312 TI - Molecular evolution of FtsZ protein sequences encoded within the genomes of archaea, bacteria, and eukaryota. AB - The FtsZ protein is a polymer-forming GTPase which drives bacterial cell division and is structurally and functionally related to eukaryotic tubulins. We have searched for FtsZ-related sequences in all freely accessible databases, then used strict criteria based on the tertiary structure of FtsZ and its well characterized in vitro and in vivo properties to determine which sequences represent genuine homologues of FtsZ. We have identified 225 full-length FtsZ homologues, which we have used to document, phylum by phylum, the primary sequence characteristics of FtsZ homologues from the Bacteria, Archaea, and Eukaryota. We provide evidence for at least five independent ftsZ gene duplication events in the bacterial kingdom and suggest the existence of three ancestoral euryarchaeal FtsZ paralogues. In addition, we identify "FtsZ-like" sequences from Bacteria and Archaea that, while showing significant sequence similarity to FtsZs, are unlikely to bind and hydrolyze GTP. PMID- 14743313 TI - The absence of TIR-type resistance gene analogues in the sugar beet (Beta vulgaris L.) genome. AB - The majority of known plant resistance genes encode proteins with conserved nucleotide-binding sites and leucine-rich repeats (NBS-LRR). Degenerate primers based on conserved NBS-LRR motifs were used to amplify analogues of resistance genes from the dicot sugar beet. Along with a cDNA library screen, the PCR screen identified 27 genomic and 12 expressed NBS-LRR RGAs (nlRGAs) sugar beet clones. The clones were classified into three subfamilies based on nucleotide sequence identity. Sequence analyses suggested that point mutations, such as nucleotide substitutions and insertion/deletions, are probably the primary source of diversity of sugar beet nlRGAs. A phylogenetic analysis revealed an ancestral relationship among sugar beet nlRGAs and resistance genes from various angiosperm species. One group appeared to share the same common ancestor as Prf, Rx, RPP8, and Mi, whereas the second group originated from the ancestral gene from which 12C1, Xa1, and Cre3 arose. The predicted protein products of the nlRGAs isolated in this study are all members of the non-TIR-type resistance gene subfamily and share strong sequence and structural similarities with non-TIR-type resistance proteins. No representatives of the TIR-type RGAs were detected either by PCR amplification using TIR type-specific primers or by in silico screening of more than 16,000 sugar beet ESTs. These findings suggest that TIR type of RGAs is absent from the sugar beet genome. The possible evolutionary loss of TIR type RGAs in the sugar beet is discussed. PMID- 14743314 TI - Likelihood analysis of the chalcone synthase genes suggests the role of positive selection in morning glories (Ipomoea). AB - Chalcone synthase (CHS) is a key enzyme in the biosynthesis of flavonoides, which are important for the pigmentation of flowers and act as attractants to pollinators. Genes encoding CHS constitute a multigene family in which the copy number varies among plant species and functional divergence appears to have occurred repeatedly. In morning glories (Ipomoea), five functional CHS genes (A E) have been described. Phylogenetic analysis of the Ipomoea CHS gene family revealed that CHS A, B, and C experienced accelerated rates of amino acid substitution relative to CHS D and E. To examine whether the CHS genes of the morning glories underwent adaptive evolution, maximum-likelihood models of codon substitution were used to analyze the functional sequences in the Ipomoea CHS gene family. These models used the nonsynonymous/synonymous rate ratio (omega = d(N)/ d(S)) as an indicator of selective pressure and allowed the ratio to vary among lineages or sites. Likelihood ratio test suggested significant variation in selection pressure among amino acid sites, with a small proportion of them detected to be under positive selection along the branches ancestral to CHS A, B, and C. Positive Darwinian selection appears to have promoted the divergence of subfamily ABC and subfamily DE and is at least partially responsible for a rate increase following gene duplication. PMID- 14743315 TI - Phylogenetic relationships among East African haplochromine fish as revealed by short interspersed elements (SINEs). AB - Genomic DNA libraries were prepared from two endemic species of Lake Victoria haplochromine (cichlid) fish and used to isolate and characterize a set of short interspersed elements (SINEs). The distribution and sequences of the SINEs were used to infer phylogenetic relationships among East African haplochromines. The SINE-based classification divides the fish into four groups, which, in order of their divergence from a stem lineage, are the endemic Lake Tanganyika flock (group 1); fish of the nonendemic, monotypic, widely distributed genus Astatoreochromis (group 2); the endemic Lake Malawi flock (group 3); and group 4, which contains fish from widely dispersed East African localities including Lakes Victoria, Edward, George, Albert, and Rukwa, as well as many rivers. The group 4 haplochromines are characterized by a subset of polymorphic SINEs, each of which is present in some individuals and absent in others of the same population at a given locality, the same morphologically defined species, and the same mtDNA defined haplogroup. SINE-defined group 4 contains six of the seven previously described mtDNA haplogroups. One of the polymorphic SINEs appears to be fixed in the endemic Lake Victoria flock; four others display the presence-or-absence polymorphism within the species of this flock. These findings have implications for the origin of Lake Victoria cichlids and for their founding population sizes. PMID- 14743316 TI - Evolutionary relationships in the sand-dwelling cichlid lineage of lake tanganyika suggest multiple colonization of rocky habitats and convergent origin of biparental mouthbrooding. AB - The cichlid species flock of Lake Tanganyika is comprised of seven seeding lineages that evolved in step with changes of the lake environment. One seeding lineage diversified into at least six lineages within a short period of time. Our study focuses on the diversification of one of these lineages, the Ectodini, comprising highly specialized, sand- and rock-dwelling species. They display two distinct breeding styles: maternal and biparental mouthbrooding. By analyzing three mtDNA gene segments in 30 species representing all 13 described genera, we show that the Ectodini rapidly diversified into four clades at the onset of their radiation. The monotypic genus Grammatotria is likely to represent the most ancestral split, followed by the almost contemporary origin of three additional clades, the first comprising the benthic genus Callochromis, the second comprising the benthic genera Asprotilapia, Xenotilapia, Enantiopus, and Microdontochromis, and the third comprising the semi-pelagic genera Ophthalmotilapia, Cardiopharynx, Cyathopharynx, Ectodus, Aulonocranus, Lestradea, and Cunningtonia. Our study confirms the benthic and sand-dwelling life-style as ancestral. Rocky habitats were colonized independently in the Xenotilapia- and Ophthalmotilapia-clade. The Xenotilapia-clade comprises both maternal and biparental mouthbrooders. Their mode of breeding appears to be highly plastic: biparental mouthbrooding either evolved once in the common ancestor of the clade, to be reverted at least three times, or evolved at least five times independently from a maternally mouthbrooding ancestor. Furthermore, the genera Xenotilapia, Microdontochromis, Lestradea, and Ophthalmotilapia appeared paraphyletic in our analyses, suggesting the need of taxonomic revision. PMID- 14743317 TI - Genetic variation in a cline in a living intertidal snail arose in the Neogastropoda over 100 million years ago. AB - To understand how species adapt and evolve it is necessary to appreciate the relationship between genetic variation and the environment. Here, the fossil record and molecular data from different lineages of the marine Gastropoda are used to understand the evolution of genetic variations found in the nuclear gene for mitochondrial malate dehydrogenase (mMDH; EC 1.1.1.37) in the living intertidal Muricid snail, Nucella lapillus. Assuming a molecular clock, DNA sequences of mMDH indicate that two variants found in N. lapillus, mMDH(9) and mMDH(10), may have arisen as long as 144 MY (million years) ago and at least prior to the evolution of the Muricidae approximately 112-90 MY ago. The Muricidae contain by far the greatest majority of the Neogastropoda specialized for life in the intertidal habitat. In N. lapillus the mMDH(9) and mMDH(10) variants covary with variations in other biochemically defined loci, inherited phenotypic traits (shell shape and physiology) and karyotype frequencies to differentiate two distinct nuclear haplotypes that are associated with different temperature environments. The variations in shell shape that are associated with the haplotypes of N. lapillus represent adaptations to temperature stress and similar variations occur in other related intertidal molluscs whose lineages are much older than Nucella, which arose around 25 million years ago. It is suggested that the divergence of the mMDH variants found in N. lapillus may reflect an ancient genetic event, such as a chromosomal mutation, perhaps involving variation in other linked traits that together became important in the subsequent evolution of the marine Gastropoda. PMID- 14743318 TI - Novel neuropeptide Y Y2-like receptor subtype in zebrafish and frogs supports early vertebrate chromosome duplications. AB - The Y receptors comprise a family of G-protein coupled receptors with neuropeptide Y-family peptides as endogenous ligands. The Y receptor family has five members in mammals and evolutionary data suggest that it diversified in the two genome duplications proposed to have occurred early in vertebrate evolution. If this theory holds true, it allows for additional family members to be present. We describe here the cloning, pharmacological characterization, tissue distribution, and chromosomal localization of a novel subtype of the Y-receptor family, named Y7, from the zebrafish. We also present Y7 sequences from rainbow trout and two amphibians. The new receptor is most similar to Y2, with 51-54% identity. As Y2 has also been cloned from some of these species, there clearly are two separate Y2-subfamily genes. Chromosomal mapping in zebrafish supports origin of Y7 as a duplicate of Y2 by chromosome duplication in an early vertebrate. Y7 has probably been lost in the lineage leading to mammals. The pharmacological profile of the zebrafish Y7 receptor is different from mammalian Y2, as it does not bind short fragments of NPY with a high affinity. The Y7 receptor supports the theory of early vertebrate genome duplications and suggests that the Y family of receptors is a result of these early genome duplications. PMID- 14743319 TI - The rDNA ITS region in the lessepsian marine angiosperm Halophila stipulacea (Forssk.) Aschers. (Hydrocharitaceae): intragenomic variability and putative pseudogenic sequences. AB - Halophila stipulacea is a dioecious marine angiosperm, widely distributed along the western coasts of the Indian Ocean and the Red Sea. This species is thought to be a Lessepsian immigrant that entered the Mediterranean Sea from the Red Sea after the opening of the Suez Canal (1869). Previous studies have revealed both high phenotypic and genetic variability in Halophila stipulacea populations from the western Mediterranean basin. In order to test the hypothesis of a Lessepsian introduction, we compare genetic polymorphism between putative native (Red Sea) and introduced (Mediterranean) populations through rDNA ITS region (ITS1-5.8S ITS2) sequence analysis. A high degree of intraindividual variability of ITS sequences was found. Most of the intragenomic polymorphism was due to pseudogenic sequences, present in almost all individuals. Features of ITS functional sequences and pseudogenes are described. Possible causes for the lack of homogenization of ITS paralogues within individuals are discussed. PMID- 14743320 TI - Different rates of LINE-1 (L1) retrotransposon amplification and evolution in New World monkeys. AB - LINE-1 (L1) elements constitute the major family of retrotransposons in mammalian genomes. Here we report the first investigation of L1 evolution in New World monkeys (NWM). Two regions of the second open-reading frame were analyzed by two methods in three NWM species, the squirrel monkey (Saimiri sciureus), the tamarin (Saguinus oedipus), and the spider monkey (Ateles paniscus). Since these three species diverged, L1 has amplified in the Saimiri and Saguinus lineages but L1 activity seems to have been strongly reduced in the Ateles lineage. In addition, the active L1 lineage has evolved rapidly in Saimiri and Saguinus, generating species-specific subfamilies. In contrast, we found no evidence for a species specific subfamily in Ateles, a result consistent with the low L1 activity in this species for the last approximately 25 My. PMID- 14743321 TI - Urothelial mesh--a new technique of cell culture on biomaterials. AB - INTRODUCTION: Urogenital malformations, trauma or tumours may demand surgical reconstruction in children. Cell culture is an important technology in biomaterial research and tissue engineering. Tissue-engineering of urothelial organs is of interest in children, because the number of complications and re operations may be reduced. Actually, monolayer cultures of urothelium are used for tissue engineering and biocompatibility testing. A culture system that more closely mimics the physiologic environment of the urothelium would be of interest. The aim of this study was to determine the biological and mechanical characteristics of urothelial mesh cultured in vitro. METHODS: Meshes containing urothelium, lamina propria, and submucosal tissue were generated using a skin mesh graft cutter. Meshes were cultured in 6-well plates, on collagen I/III, polydioxanone/polylactic acid and silicone matrices. Cell morphology was examined by inversion microscopy, histology, and scanning electron microscopy. It was compared to urothelium cultured by methods reported in the literature. To define the basic mechanical properties, meshes were extended longitudinally by a servohydraulic testing machine and strain diagrams generated. RESULTS: Urothelium was reproducibly cultured from meshes. Cell growth could be induced onto fibrillary collagen, polydioxanone-polylactic acid matrices and shaped polyurethane surfaces. Cells formed confluent layers of flat cells, resembling native urothelium. The meshes have unique mechanical properties, allowing for stable fixation, surgical handling and mechanical stimulation. CONCLUSIONS: Meshes can be used for cell culture on biomaterials. They maintain epithelial stromal integrity and mechanic stability. The small size of tissue bridges allows in vitro culture for long periods with many potential advantages for tissue engineering and biologic research. Applications are possible both in vitro and in vivo. PMID- 14743322 TI - Coexisting pelviureteral and vesicoureteral junction obstruction in children. AB - OBJECTIVE: To address reasons for diagnostic failure in children with coexisting pelviureteral and vesicoureteral junction obstruction. PATIENTS AND METHODS: A total of 11 children with coexisting pelviureteral and vesicoureteral junction obstruction are reported. This anomaly was recognised at presentation in only 5 (45%) of cases. In the remainder, correct diagnosis was made after initial pyeloplasty and a significant delay occurred in 3 children. In all 11 patients treatment consisted of pyeloplasty and subsequent reimplantation. RESULTS: A satisfactory postoperative course with improvement of urinary tract dilatation was noted in all 11 patients, while 8 of 11 patients showed preserved renal function at late follow-up. CONCLUSION: According to the few available data, our data confirm the difficulty of diagnosing coexisting pelviureteral and vesicoureteral junction obstruction in children. However, prompt recognition and relief of urinary obstruction in these patients is mandatory in order to prevent deterioration of renal function. PMID- 14743323 TI - Urachal remnants: an enigma. AB - AIM: To determine what should be done with incidentally discovered urachal remnants. MATERIALS AND METHODS: A 10-year retrospective study revealed 24 patients with urachal remnants. The histology of the urachal remnants was reviewed. A review of the data on urachal carcinoma in adults diagnosed and treated during the same period in an adjacent adult hospital was carried out. A review of the literature was also performed. RESULTS: In the 23 patients where histology was available 17 had normal urothelial lining and 6 (25%) showed abnormal epithelium. This included colonic epithelium, small intestinal epithelium and squamous epithelium. Four of the patients with abnormal epithelium were in the group of "incidental removals". During the 10-year period, 4 cases of urachal carcinoma were managed at the adult urology centre. Three had extensive disease at presentation. Two died of the disease while the third died of myocardial infarction 5 months after resection. Only one was alive at one-year follow-up. The occurrence of such documented complications makes a strong case for "prophylactic" surgery when urachal remnants are diagnosed incidentally or recognised during other operative procedures. All paediatric patients who underwent excision of urachal remnants had an uncomplicated clinical course. PMID- 14743324 TI - Exstrophy variants: should they be considered malformation complexes separate from classic exstrophy? AB - PURPOSE: Exstrophy variants are very rare and have a better prognosis than classical exstrophy. The authors came across a case of superior vesical fissure (SVF) together with esophageal atresia and tracheoesophageal fistula (EATEF) and a case of SVF with gross limb anomalies. These associated malformations have not been reported so far in the literature and hence we reviewed all the cases of exstrophy variants presented to us with particular emphasis on the associated malformations. METHODS: Records (n=9) of patients who were diagnosed as exstrophy variants at our institution between 1989 and 2000 were evaluated retrospectively. RESULTS: Out of 9 cases, 7 cases had associated malformations: EATEF, urethral atresia, absent radius, large umbilical hernia, low anorectal malformation, true diphallus with bifid scrotum, or high anorectal malformation. CONCLUSION: The high incidence of associated congenital malformations, noted in our exstrophy variant series, raises doubts about the clubbing together of the exstrophy variants with classical exstrophy. Further investigation of such cases may elucidate shared or unique causes of the dysembryogenic mechanisms in the etiologies of variants of bladder exstrophy. PMID- 14743325 TI - Combined use of perimeatal-based flap urethroplasty (Mathieu) with midline incision or urethral plate in hypospadias repair. AB - Perimeatal-based flap urethroplasty is commonly used for the primary correction of distal hypospadias. The Mathieu repair provides excellent function and satisfactory cosmetic results, but the risk of devascularisation of the neourethral flap is an inherent problem of this technique. The midline incision of the urethral plate is part of the Snodgrass method, which offers good urethral calibre and tension-free closure. Our aim was to test the effectiveness of the use of the perimeatal-based flap combined with a midline incision of the urethral plate in hypospadias repair. 19 boys, aged 20 months to 5 years, with distal hypospadias, underwent a one-stage repair using the above-described modification of the Mathieu technique. The meatal-based flap with a midline incision of the urethral plate was similar in all cases. The hypospadias repair was successful in all patients, only one boy developed a urethrocutaneous fistula which required subsequent operation. There was no postoperative scarring, chordee or urethral stricture during follow-up. Hypospadias repair using a perimeatal-based skin flap and combined with an incision of the urethral plate reduces complication rates and offers good cosmetic results. PMID- 14743326 TI - Critical validation of colour Doppler ultrasound in diagnostics of acute scrotum in children. AB - Acute scrotum in children is generally a symptom. Several pathologies of the testis, scrotum or groin may cause it, including inflammatory as well as ischaemic processes. Testicular torsion with ischaemia of the testis is an emergency requiring prompt surgical intervention. Even experienced paediatric surgeons and urologists may have difficulties differentiating an ischaemic from an inflammatory aetiology solely on the basis of clinical signs and symptoms. In cases of aetiologic uncertainty, operative exploration has to be performed to rule out ischaemia, which led in the past to a high rate of surgery. More recently, colour Doppler ultrasonography (CDUS) has become an imaging modality for the examination of the acute scrotum with the purpose of detecting ischaemia, thus reducing the need for explorative surgery. In this study we examined the reliability of CDUS, comparing the sonographical diagnoses with both the intraoperative findings and the clinical course. We present here 132 consecutive cases of acute scrotum in children, who were admitted from 01/98 through 10/02 to our hospital. All patients underwent CDUS. Patients with ultrasonographic signs of ischaemia of the testis (group A) and patients with demonstrated perfusion and suspicious or severe clinical symptoms (group B) were operated immediately. Patients with demonstrated perfusion without suspicious or severe symptoms (group C) were treated conservatively. Thirty-eight patients (28.8%) were treated operatively. Eleven cases of testicular torsion in group A (12 pat.) could be confirmed intraoperatively, in one case there was no testicular torsion but a severe inflammation of the testis with torsion of the appendix testis. In group B (26 pat.) operative exploration revealed one case of testicular torsion. In group B two cases of sonographically suspected torsions of the appendix testis also emerged as epididymitis intraoperatively. In the other cases postoperative diagnosis was in agreement with the preoperative CDUS. The clinical course of 94 conservatively treated patients (71.2%) (group C) confirmed in all cases a non ischaemic aetiology. There was a total of 12 cases (9.1%) where acute scrotum was caused by testicular torsions. This relatively low rate underlines the necessity to differentiate between ischaemic and other aetiologies, thus avoiding operation in many patients. 71.2% of our patients could be treated conservatively. However, it is essential to exclude an ischaemic cause before opting for conservative therapy. Even though CDUS is a very valuable diagnostic tool in this sense, this exclusion cannot be ensured in every case and is highly dependent on the expertise and technique of the investigator. Therefore in cases with any doubts an operative exploration has to be performed to avoid the most dreaded complication in the treatment of acute scrotum. PMID- 14743327 TI - Treatment of post-appendectomy intra-abdominal deep abscesses. AB - The treatment of acute appendicitis in children is sometimes followed by complications including intra-abdominal abscess, for which the traditional treatment is surgical drainage. We evaluated the efficacy of antibiotic management compared to classic surgical treatment. This retrospective study investigated 22 children from 5 to 13 years of age with one or many abscesses after appendectomy, treated between 1992 and 2002. Eleven received surgery and the other 11 were treated with triple antibiotherapy. The two groups were comparable. Surgery was efficient in 36% of cases and complications occurred in 64% of cases (digestive fistula, intraperitoneal abscess, gaseous gangrene and septic shock). Average hospital stay in this group was 16.7 days. In the other group, medication was efficient in 91% of cases; a recurrent abscess was operated and a residual stercolith, which was maintaining the infection, was removed. The average hospital stay in this group was 10.4 days. Medical treatment of intraperitoneal abscess seems to be effective. Hospitalisation is shorter with medical management and complications are rare. Therefore, medication may be proposed in most cases, except when there are residual foreign bodies or stercoliths. Poor patient status and septic shock are the two other contraindications, because antibiotherapy is not immediately efficient. PMID- 14743328 TI - Tracheobronchial injury by blunt trauma in children: is emergency tracheobronchoscopy always necessary? AB - PURPOSE: To discuss the usefulness of CT scan in initial management of well tolerated tracheobronchial injuries and the place of tracheoscopy. METHODS: We report our experience of three cases treated for tracheobronchial rupture resulting from three different mechanisms and review the literature. Three boys, aged 4 to 10 years, were referred to our institution for tracheobronchial rupture. Two of them presented with subcutaneous emphysema after a minor trauma, the third was a polytrauma referred after a severe car crash and was already intubated. We emphasise the importance of an initial CT scan, as this allowed us twice to confirm the tracheal wound prior to tracheoscopy. We discuss the necessity of performing a tracheoscopy in the case of a well tolerated lesion, as this procedure clearly worsened the ventilatory state in one of our cases. Moreover, one of our cases illustrates the fact that even a minor trauma can lead to life-threatening respiratory distress. All the lesions observed in our study were linear and were managed by thoracic drainage; they were then closely monitored and required no further surgical procedure. CONCLUSION: Tracheobronchial rupture in children can result from minor cervical traumas and in such cases special attention must be paid to mild discomfort or subcutaneous emphysema on admission. Initial CT scan can be very helpful in visualising the level of the rupture and its consequences with respect to the pulmonary parenchyma. One can question the necessity for tracheoscopy in well tolerated lesion, as its results do not always improve the therapeutic outcome. PMID- 14743329 TI - Recurrent immature mediastinal teratoma with life-threatening respiratory distress in a neonate. AB - This case report describes a newborn with a mediastinal teratoma who presented with severe respiratory distress at birth and required emergency surgery. The mass was adherent to the pericardium and the aorta and was compressing the trachea. Histopathology proved it to be an immature teratoma. After the excision, the elevated serum alpha fetoprotein (AFP) levels dropped to normal. However, at 4 months of age the AFP level started to rise again and a recurrent lesion was detected. This was treated with Carboplatin-based chemotherapy. The child responded favorably and is well after 42 months' follow-up. Of the few patients presenting with a similar diagnosis, none recurred nor any malignancy reported. We have reviewed the literature of this clinical rarity. PMID- 14743330 TI - Congenital left-sided Bochdalek diaphragmatic hernia. Thoracoscopic repair--case report. AB - The authors report the case of a 12-year-old girl who presented with a left-sided diaphragmatic hernia that was diagnosed after a pulmonary infection. CT scan confirmed a posterolateral diaphragmatic Bochdalek defect. The child underwent a thoracoscopic repair of the defect in 1997. The herniated contents included spleen, stomach, and intestine. The child was discharged from hospital the day after surgery. She has been followed up and is clinical and radiologically well. PMID- 14743331 TI - Gastric outlet obstruction caused by heterotopic pancreas. AB - Heterotopic pancreas (HP) in the stomach is a relatively well-known entity, but there are not many symptomatic cases reported in children. We report on a 9-year old boy presenting with nausea and vomiting. The first gastroscopic examination showed a crater-like lesion in the antrum, but at follow-up gastroscopy a few weeks later the lesion was polypoid, obstructing the pylorus. Endoscopic biopsy was not diagnostic, but histological examination after open excision showed HP. It is unclear why the lesion changed so markedly in appearance in just a few weeks. HP is a rare cause of gastric outlet obstruction in children. PMID- 14743332 TI - Diaphragmatic duodenal atresia: laparoscopic repair. AB - Stenosis due to a diaphragm is a type of intrinsic duodenal obstruction in newborns and even in childhood, when obstruction is partial. We present a case of a 13-month-old girl with diaphragmatic stenosis associated with a dilated first and second duodenum. Surgical management consisted of a partial excision of the diaphragm after vertical incision of the anterior part of the second duodenum followed by a transverse suture. This diamond-shaped anastomosis was successfully carried out laparoscopically. No tapering of the duodenum was performed as some authors suggest in cases of megaduodenum. The rapid resumption of peristalsis and fewer adherences than expected after such a minimally invasive approach could make a more invasive procedure unnecessary. Only long-term follow-up and greater experience will show which procedure is most suitable. PMID- 14743333 TI - Interest of laparoscopy in polysplenia syndrome. AB - Polysplenia syndrome (PS) is usually discovered in symptomatic patients in association with congenital heart disease or biliary atresia. Asymptomatic patients can present associated anomalies of the digestive tract such as intestinal malrotation and gastric or splenic malfixation. We report a case of PS presenting as a left flank mass shown to be an accessory spleen by denatured red blood cell scintigraphy. Upper gastrointestinal tract studies showed intestinal malrotation. Laparoscopic exploration confirmed intestinal malrotation and showed the absence of fixation of the accessory spleen. We performed a laparoscopic Ladd's procedure followed by fixation of the accessory spleen and resection of Meckel's diverticulum through a short left-flank incision. PMID- 14743334 TI - Inflammatory myofibroblastic tumour of the extrahepatic bile ducts: an unusual cause of obstructive jaundice in children. AB - Inflammatory myofibroblastic tumour is a rare entity in children with few reported series. The extrahepatic bile duct is an unusual location for this tumour. The authors report here the case of an 8-year-old girl presenting with obstructive jaundice due to inflammatory myofibroblastic tumour of the extrahepatic bile ducts with differential diagnosis of obstructive jaundice in children. PMID- 14743335 TI - Chilaiditi syndrome associated with transverse colon volvulus: first report in a paediatric patient and review of the literature. AB - A hepatodiaphragmatic interposition of the colon, known as Chilaiditi's sign, is usually discovered by chance during the study of another event, given that its presentation is normally asymptomatic. When this finding is accompanied by clinical symptoms, either intermittent or persistent, it is known as Chilaiditi syndrome. It may be associated with intestinal obstruction due to twisting. The association of Chilaiditi syndrome and transverse colon volvulus is exceptional. To date only three cases have been reported, all in adult males. Among the common predisposing factors were anatomical alterations of the intestine such as elongation of the colon and a history of prior abdominal surgery. The clinical symptoms were due to the intestinal obstruction. We present the first description in the paediatric population of an association of transverse colon volvulus and Chilaiditi syndrome whose predisposing factors, clinical symptoms and treatment differed from those reported in the non-paediatric cases published to date. PMID- 14743339 TI - The journal celebrates an anniversary. PMID- 14743340 TI - Genetic damage detected in CD-1 mouse pups exposed perinatally to 3'-azido-3' deoxythymidine and dideoxyinosine via maternal dosing, nursing, and direct gavage. AB - Human immunodeficiency virus (HIV)-infected pregnant women are administered nucleoside-analogue antiretrovirals to reduce maternal-infant viral transmission. The current protocol recommends treating newborns for 6 additional weeks postpartum. The treatment is effective, but the risk of drug-induced chromosomal damage in neonates remains undefined. We used a mouse model to investigate this concern. In a multigeneration reproductive toxicity study, female CD-1 mice received 3'-azido-3'-deoxythymidine (AZT) and dideoxyinosine (ddI) (50/250, 75/375, 150/750 mg/kg/day AZT/ddI) by gavage twice daily in equal fractions beginning prior to mating and continuing throughout gestation and lactation. Direct pup dosing (same regimen) began on postnatal day (PND) 4. Peripheral blood erythrocytes of male pups were screened for micronuclei, markers of chromosomal damage, on PNDs 1, 4, 8, and 21. Extraordinary increases in micronucleated cells were noted in pups for each treatment group at each sampling time; treated dams exhibited smaller yet significant increases in micronucleated erythrocytes. The frequencies of micronucleated cells in untreated pups were higher than in the untreated dams, and all pups had markedly elevated levels of circulating reticulocytes compared to dams. These observations suggest that fetal and neonatal mouse hematopoietic precursor cells have heightened sensitivity to genotoxic agents, perhaps due to rapid cell proliferation during the perinatal period of development. The amount of genetic damage observed in treated pups raises concern for the potential of similar damage in humans. Investigations of chromosomal integrity in exposed newborns and children are recommended. PMID- 14743341 TI - Use of a high-throughput umu-microplate test system for rapid detection of genotoxicity produced by mutagenic carcinogens and airborne particulate matter. AB - In the present study, we developed a rapid umu-microplate test system that uses the nitroreductase- and O-acetyltransferase-overproducing Salmonella typhimurium strain NM3009 and the O-acetyltransferase-overproducing S. typhimurium strain NM2009 to detect genotoxic activity in small volume samples. The assay was used to test the genotoxicity of several standard mutagens and environmental samples. Exponentially growing cultures of NM3009, NM2009, and the parental strain TA1535/pSK1002 were incubated in 96-well microplates with test chemicals both in the presence and in the absence of rat liver S9. The relative beta-galactosidase activities were then determined colorimetrically using either chlorophenol red beta-D-galactopyranoside (CPRG) or O-nitrophenyl-beta-D-galactopyranoside (ONPG) as a measure of umuC gene induction activity. The sensitivities of NM3009 without S9 mix and NM2009 with S9 mix to nitroarenes and aromatic amines were up to 24- to 75-fold higher than those of the parent strain. Induction of umuC gene expression was detected more readily with CPRG than ONPG. The umu-microplate assay also detected genotoxicity in organic extracts of particulate matter from air samples collected in Osaka City, Japan. The pattern of the responses suggested that the genotoxic activity of the particulate extract was due primarily to nitrated polycyclic aromatic hydrocarbons. Our results indicate that the umu-microplate assay may be a useful way of carrying out rapid screens for genotoxicity in small-volume environmental samples. PMID- 14743342 TI - Comparative genomic hybridization analysis of N-methyl-N'-nitrosoguanidine induced rat gastrointestinal tumors discloses a cytogenetic fingerprint. AB - Exposure to N-nitroso compounds is thought to play a key role in the development of gastric cancer in humans. The alkylating agent N-methyl-N'-nitrosoguanidine (MNNG) is carcinogenic in a number of animal models and its preferential target tissue is the gastrointestinal (GI) tract. The genetic synteny among rats and humans makes the rat a useful model for induced tumorigenesis. However, because of the limited availability of genetic information, cytogenetic and molecular studies are rarely performed in the rat. We report an investigation of eight MNNG induced rat gastric tumors by comparative genomic hybridization (CGH). The tumors were from forestomach (induced by a single dose of MNNG) and from pylorus (induced by chronic exposure). CGH identified a genetic fingerprint of chromosomal imbalances common to the two types of the tumors. Frequent gains were observed at 9q11-q12, 15q22-25, and Xq11-q12. Forestomach carcinomas were also characterized by gains in 7q11-q12, 20q13, and Yq12. Homology studies between the rat and human genomes indicate the presence of genes within these regions with potential relevance to tumorigenesis in the GI tract. Our findings provide new insights into the location of genes involved in MNNG-induced gastric cancer initiation and/or progression in the rat. PMID- 14743343 TI - Mutation induction in haploid yeast after split-dose radiation exposure. II. Combination of UV-irradiation and X-rays. AB - Split-dose protocols can be used to investigate the kinetics of recovery from radiation damage and to elucidate the mechanisms of cell inactivation and mutation induction. In this study, a haploid strain of the yeast, Saccharomyces cerevisiae, wild-type with regard to radiation sensitivity, was irradiated with 254-nm ultraviolet (UV) light and then exposed to X-rays after incubation for 0-6 hr. The cells were incubated either on nutrient medium or salt agar between the treatments. Loss of reproductive ability and mutation to canavanine resistance were measured. When the X-ray exposure immediately followed UV-irradiation, the X ray survival curves had the same slope irrespective of the pretreatment, while the X-ray mutation induction curves were changed from linear to linear quadratic with increasing UV fluence. Incubations up to about 3 hr on nutrient medium between the treatments led to synergism with respect to cell inactivation and antagonism with respect to mutation, but after 4-6 hr the two treatments acted independently. Incubation on salt agar did not cause any change in the survival curves, but there was a strong suppression of X-ray-induced mutation with increasing UV fluence. On the basis of these results, we suggest that mutation after combined UV and X-ray exposure is affected not only by the induction and suppression of DNA repair processes, but also by radiation-induced modifications of cell-cycle progression and changes in the expression of the mutant phenotype. PMID- 14743344 TI - Population doubling: a simple and more accurate estimation of cell growth suppression in the in vitro assay for chromosomal aberrations that reduces irrelevant positive results. AB - International guidelines for cytotoxicity limits for the in vitro chromosomal aberration assay require reductions in cell growth of greater than 50%. This sets no upper limit on toxicity and there is concern about the number of false or irrelevant results obtained in the aberration assay, i.e., positive results at toxic dose levels only, with no evidence for primary DNA damaging ability and with negative results in the other genotoxicity tests. We have previously proposed that no truly genotoxic compound would be missed if the toxicity of the highest dose did not exceed 50%. Cell growth measured by cell counts as a percentage of controls can underestimate toxicity. For example, if we seed half a million cells per culture, and the controls double to 1 million during the experiment, a culture that truly has no growth will still have a cell count 50% of the control. Measurement of population doublings (PDs) more accurately assesses cell growth. To assess the use of PD in dose selection, we examined previous data from this lab and data from new experiments with "true," primary DNA damaging clastogens, and with clastogens, including drugs, thought to act indirectly, through cytotoxicity-associated mechanisms. We compared aberration results where the highest doses scored were based on 50% reductions in final cell counts with results obtained when the highest doses were based on PD. The PD method allows detection of true clastogens, including those that are active in a range with some toxicity, and reduces the number of toxicity-related "false" positive results. PMID- 14743345 TI - Effect of cyanidin 3-O-beta-glucopyranoside on micronucleus induction in cultured human lymphocytes by four different mutagens. AB - The anthocyanin cyanidin 3-O-beta-glucopyranoside (Cy-g) is reported to be one of the most effective antioxidants, but little is currently known regarding its potential chemopreventive properties. In this study, we evaluated the ability of Cy-g to protect cultured human lymphocytes from micronucleus (MN) induction by four different mutagens: ethyl methanesulfonate (EMS), colchicine (COL), H(2)O(2), and mitomycin C (MMC). To gain insight into the mechanisms of action of Cy-g, the cultures were treated with the compound before, during, and after treatment with the mutagens; in addition, the cultures were evaluated for the induction of apoptosis. When used by itself, up to 100 microg/ml of Cy-g was nongenotoxic, while 100 microg/ml Cy-g reduced the replicative index of the cells by nearly 50%. In addition, Cy-g was able to reduce the frequency of micronuclei induced by EMS, COL, and H(2)O(2) using all three treatment protocols, but it had no significant effect on MN induction by MMC in any of the protocols. Apoptosis was produced in the cultures treated with Cy-g alone and was increased under conditions in which Cy-g produced anti-genotoxic effects, suggesting that Cy-g mediated-apoptosis may remove highly damaged cells. However, increases in apoptosis were found under conditions in which Cy-g was not significantly anti genotoxic, indicating that the increases in apoptosis were not sufficient to account for the anti-genotoxicity of Cy-g. Taken together, our findings indicate that Cy-g possesses anti-genotoxic activity in vitro, which suggests its potential use as a chemopreventive agent. PMID- 14743346 TI - Urinary mutagenesis and fried red meat intake: influence of cooking temperature, phenotype, and genotype of metabolizing enzymes in a controlled feeding study. AB - Meat cooked at high temperatures contains potential carcinogenic compounds, such as heterocyclic amines (HCAs) and polycyclic aromatic hydrocarbons (PAHs). Samples from a 2-week controlled feeding study were used to examine the relationship between the intake of mutagenicity from meat fried at different temperatures and the levels of mutagenicity subsequently detected in urine, as well as the influence of the genotype of drug metabolizing enzymes on urinary mutagenicity. Sixty subjects consumed ground beef patties fried at low temperature (100 degrees C) for 1 week, followed by ground beef patties fried at high temperature (250 degrees C) the second week. Mutagenicity in the meat was assayed in Salmonella typhimurium TA98 (+S9), and urinary mutagenicity was determined using Salmonella YG1024 (+S9). Genotypes for NAT1, NAT2, GSTM1, and UGT1A1 were analyzed using blood samples from the subjects. Meat fried at 100 degrees C was not mutagenic, whereas meat fried at 250 degrees C was mutagenic (1023 rev/g). Unhydrolyzed and hydrolyzed urine samples were 22x and 131x more mutagenic, respectively, when subjects consumed red meat fried at 250 degrees C compared with red meat fried at 100 degrees C. We found that hydrolyzed urine was approximately 8x more mutagenic than unhydrolyzed urine, likely due to the deconjugation of mutagens from glucuronide. The intake of meat cooked at high temperature correlated with the mutagenicity of unhydrolyzed urine (r = 0.32, P = 0.01) and hydrolyzed urine (r = 0.34, P = 0.008). Mutagenicity in unhydrolyzed urine was not influenced by NAT1, NAT2, or GSTM1 genotypes. However, a UGT1A1*28 polymorphism that reduced UGT1A1 expression and conjugation modified the effect of intake of meat cooked at high temperature on mutagenicity of unhydrolyzed urine (P for interaction = 0.04). These mutagenicity data were also compared with previously determined levels of HCAs (measured as MeIQx, DiMeIQx, and PhIP) and polycyclic aromatic hydrocarbons (PAHs) in the meat, levels of HCAs in the urine, and CYP1A2 and NAT2 phenotypes. The levels of mutagenicity in the meat fried at low and high temperatures correlated with levels of HCAs, but not levels of PAHs, in the meat. Also, levels of mutagenicity in unhydrolyzed urine correlated with levels of MeIQx in unhydrolyzed urine (r = 0.36; P = 0.01), and the levels of mutagenicity of hydrolyzed urine correlated with levels of MeIQx (r = 0.34; P = 0.01) and PhIP (r = 0.43; P = 0.001) of hydrolyzed urine. Mutagenicity in unhydrolyzed urine was not influenced by either the CYP1A2 or NAT2 phenotype. The data from this study indicate that urinary mutagenicity correlates with mutagenic exposure from cooked meat and can potentially be used as a marker in etiological studies on cancer. PMID- 14743347 TI - Characterisation of chicken TES and its role in cell spreading and motility. AB - Previously we identified TES as a candidate tumour suppressor gene that is located at human chromosome 7q31.1. More recently, we and others have shown TES to encode a novel LIM domain protein that localises to focal adhesions. Here, we present the cloning and functional analysis of the chicken orthologue of TES, cTES. The TES proteins are highly conserved between chicken and human, showing 89% identity at the amino acid level. We show that the cTES protein localised at focal adhesions, actin stress fibres, and sites of cell-cell contact, and GST cTES can pull-down zyxin and actin. To investigate a functional role for cTES, we looked at the effect of its overexpression on cell spreading and cell motility. Cells overexpressing cTES showed increased cell spreading on fibronectin, and decreased cell motility, compared to RCAS vector transfected control cells. The data from our studies with cTES support our previous findings with human TES and further implicate TES as a member of a complex of proteins that function together to regulate cell adhesion and additionally demonstrate a role for TES in cell motility. PMID- 14743348 TI - How adhesion, migration, and cytoplasmic calcium transients influence interleukin 1beta mRNA stabilization in human monocytes. AB - We investigated the mechanisms by which primary human monocyte migration and the production of important cytokines are co-regulated. Motile monocytes underwent cyclic morphologic and adhesive changes that were associated with intracellular free calcium changes; in such cells, cytokine transcripts were unstable and translationally repressed. Agents that activate monocytes, including lipopolysacharrides (LPS), cytomegalovirus (CMV), and tumor necrosis factor (TNFalpha), have been shown to de-repress translation and these agents stabilize adhesion-induced transcripts for IL-lbeta and IL-8 and markedly diminish cell migration in the presence of autologous serum. LPS suppressed Rho A activity and either this agent or C3 transferase elevated intracellular free calcium, stabilized transcripts, and, in tandem, inhibited cell migration by preventing tail retraction, a prerequisite for cell translocation. These results, therefore, suggest that monocyte activating agents inhibit the RhoA pathway and continuously elevate intracellular calcium leading to a concomitant decrease in monocyte migration and stabilization of cytokine transcripts prior to translation. PMID- 14743349 TI - The chemotaxis defect of Shwachman-Diamond Syndrome leukocytes. AB - Shwachman-Diamond Syndrome (SDS) is a rare autosomal recessive, multisystem disorder presenting in childhood with intermittent neutropenia and pancreatic insufficiency. It is characterized by recurrent infections independent of neutropenia, suggesting a functional neutrophil defect. While mutations at a single gene locus (SBDS) appear to be responsible for SDS in a majority of patients, the function of that gene and a specific defect in SDS neutrophil behavior have not been elucidated. Therefore, employing 2D and 3D computer assisted motion analysis systems, we have analyzed the basic motile behavior and chemotactic responsiveness of individual polymorphonuclear leukocytes (PMNs) of 14 clinically diagnosed SDS patients. It is demonstrated that the basic motile behavior of SDS PMNs is normal in the absence of chemoattractant, that SDS PMNs respond normally to increasing and decreasing temporal gradients of the chemoattractant fMLP, and that SDS PMNs exhibit a normal chemokinetic response to a spatial gradient of fMLP. fMLP receptors were also distributed uniformly through the plasma membrane of SDS PMNs as in control PMNs. SDS PMNs, however, were incapable of orienting in and chemotaxing up a spatial gradient of fMLP. This unique defect in orientation was manifested by the PMNs of every SDS patient tested. The PMNs of an SDS patient who had received an allogenic hematopoietic stem cell transplant, as well as PMNs from a cystic fibrosis patient, oriented normally. These results suggest that the defect in SDS PMNs is in a specific pathway emanating from the fMLP receptor that is involved exclusively in regulating orientation in response to a spatial gradient of fMLP. This pathway must function in parallel with additional pathways, intact in SDS patients, that emanate from the fMLP receptor and regulate responses to temporal rather than spatial changes in receptor occupancy. PMID- 14743350 TI - Positioning and capture of cell surface-associated microtubules in epithelial tendon cells that differentiate in primary embryonic Drosophila cell cultures. AB - Using primary embryonic Drosophila cell cultures, we have investigated the assembly of transcellular microtubule bundles in epidermal tendon cells. Muscles attach to the tendon cells of previously undescribed epidermal balls that form shortly after culture initiation. Basal capture of microtubule ends in cultured tendon cells is confined to discrete sites that occupy a relatively small proportion of the basal cell surface. These capturing sites are associated with hemiadherens junctions that link the ends of muscle cells to tendon cell bases. In vivo, muscle attachment and microtubule capture occur across the entire cell base. The cultured tendon cells reveal that the basal ends of their microtubules can be precisely targeted to small, pre-existing, structurally well-defined cortical capturing sites. However, a search and capture targeting procedure, such as that undertaken by kinetochore microtubules, cannot fully account for the precision of microtubule capture and positioning in tendon cells. We propose that cross-linkage of microtubules is also required to zip them into apicobasally oriented alignment, progressing from captured basal plus ends to apical minus ends. This involves repositioning of apical minus ends before they become anchored to an apical set of hemiadherens junctions. The proposal is consistent with our finding that hemiadherens junctions assemble at tendon cell bases before they do so at cell apices in both cultures and embryos. It is argued that control of microtubule positioning in the challenging spatial situations found in vitro involves the same procedures as those that operate in vivo. PMID- 14743351 TI - Flagellar quiescence in Chlamydomonas: Characterization and defective quiescence in cells carrying sup-pf-1 and sup-pf-2 outer dynein arm mutations. AB - Chlamydomonas reinhardtii can use their flagella for two distinct types of movement: swimming through liquid or gliding on a solid substrate. Cells switching from swimming to gliding motility undergo a reversible flagellar quiescence. This phenomenon appears to involve the outer dynein arms, since mutants having altered outer arm beta and gamma dyneins (sup-pf-1 and sup-pf-2) show a diminished ability to quiesce. Sup-pf-1 and sup-pf-2 were originally isolated as gain-of-function mutations that suppress the flagellar paralysis resulting from radial spoke or central pair defects. Defective quiescence is also a gain-of-function phenomenon, as cells completely lacking outer arm heavy chains show a normal quiescence phenotype. These data suggest that regulation of outer arm dynein activity is essential for flagellar quiescence and furthermore that regulation of quiescence involves a signal transduction pathway that shares elements with the radial spoke/central pair system. PMID- 14743354 TI - Surgery in Tourette syndrome. AB - Tourette syndrome (TS) is a neuropsychiatric disorder with onset in early childhood. It is characterized by tics and often accompanied by disturbances in behavior, such as obsessive-compulsive disorder (OCD). In most cases, the disorder is self-limited or can be treated by medication or behavioral therapy. In a small percentage, however, symptoms are intractable to any conservative treatment. Since 1955, various attempts have been made to treat these patients through neurosurgical procedures. The target sites have been diverse and include the frontal lobe (prefrontal lobotomy and bimedial frontal leucotomy), the limbic system (limbic leucotomy and anterior cingulotomy), the thalamus, and the cerebellum. Combined approaches have also been tried such as anterior cingulotomies plus infrathalamic lesions. The results have often been unsatisfactory or major side effects have occurred, such as hemiplegia or dystonia. Our review of the literature from 1960 until 2003 revealed 21 reports and 3 descriptions in textbooks covering about 65 patients in total who had undergone ablative procedures for intractable TS, the first being reported in 1962. In 1999, deep brain stimulation (DBS) was introduced as a new approach for intractable TS. To date, 3 patients have been reported who underwent bilateral thalamic stimulation, with promising results on tics and obsessive-compulsive symptoms. PMID- 14743355 TI - The monoamine reuptake inhibitor BTS 74 398 fails to evoke established dyskinesia but does not synergise with levodopa in MPTP-treated primates. AB - Long-term treatment of Parkinson's disease (PD) with levodopa (L-dopa) induces dyskinesia that, once established, is provoked by each dose of L-dopa or a dopamine (DA) agonist. In contrast, monoamine reuptake inhibitors may reverse motor deficits in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-treated primates without provoking established involuntary movements. We now examine whether the potent monoamine reuptake blocker BTS 74 398 induces established dyskinesia in MPTP-treated common marmosets primed previously with L-dopa and whether co-administration of BTS 74 398 with L-dopa potentiates motor behaviour and dyskinesia induced by acute L-dopa treatment. Administration of BTS 74 398 (2.5, 5.0, or 10.0 mg/kg, p.o.) in MPTP-treated common marmosets increased locomotor activity and reduced motor disability in a dose-related manner but did not provoke involuntary movements. BTS 74 398 (2.5, 5.0, or 10.0 mg/kg p.o.) co administered with a threshold dose of L-dopa (2.5 mg/kg p.o.) did not evoke a motor response or induce dyskinesia. Similarly, concomitant administration of BTS 74 398 (5.0 mg/kg p.o.) with a submaximal L-dopa dose (12.5 mg/kg p.o.) did not potentiate the motor response produced by L-dopa alone and there was no alteration in the dyskinesia provoked by L-dopa challenge. BTS 74 398 reverses motor abnormalities in MPTP-treated marmosets without evoking established dyskinesia but no additive improvement occurs when administered in combination with L-dopa. The lack of synergy with L-dopa may suggest different sites of drug action. PMID- 14743356 TI - Quality of life in early Parkinson's disease: impact of dyskinesias and motor fluctuations. AB - The impact of dyskinesias and motor fluctuations on quality of life (QOL) at various stages in the course of Parkinson's disease (PD) is not well understood. In 301 subjects with early PD enrolled in a clinical trial (CALM-PD), we quantified the impact of motor complications on QOL and investigated how this changes over time. We also compared QOL related to demographic and treatment characteristics. The presence of dyskinesias was associated with visual analogue scale (VAS) scores 3.0 of 100 points higher (better) than those without dyskinesias in years 1 to 2, even when adjusting for Unified Parkinson's Disease Rating Scale (UPDRS) motor scores. The positive association between dyskinesias and QOL scores was more marked in older patients. In years 3 to 4, dyskinesias no longer had a significant relationship with QOL. Younger subjects had higher VAS scores. Gender, motor fluctuations, and treatment regimen had no significant association with QOL, although a trend was found toward a small negative effect of motor fluctuations on QOL. We conclude that motor complications that occur within the first 4 years of treatment of PD do not have a significant negative effect on quality of life as measured by a visual analogue scale for most patients. PMID- 14743357 TI - Quetiapine improves psychotic symptoms and cognition in Parkinson's disease. AB - Twenty-nine elderly patients who failed treatment with clozapine, risperidone, or olanzapine entered this 24-week, single-center, open-label trial to assess the efficacy of quetiapine (12.5-400 mg/day) for psychosis in patients with Parkinson's disease (PD). Psychiatric, motor, and cognitive assessments were administered at baseline and at periodic intervals for 24 weeks. These included the Brief Psychiatric Rating Scale (BPRS), Neuropsychiatric Inventory (NPI), Unified Parkinson's Disease Rating Scale (UPDRS) and tests of intellectual functioning, attention, and memory. Repeated measures statistical analysis was used to assess change from baseline. The results revealed significant improvements in the 24-week BPRS total score and NPI psychosis subscale scores, with no decline in UPDRS total or motor subscale scores. There was also significant improvement in recall scores on cognitive measures. These results indicate that quetiapine may treat psychotic symptoms and improve cognition without worsening motor function in patients with PD, suggesting that quetiapine is an effective and well-tolerated antipsychotic in this population. PMID- 14743359 TI - Long-term outcome of focal dystonia in string instrumentalists. AB - This study describes the clinical characteristics and long-term outcome in string instrumentalists with focal task-specific dystonia. We present the results of a follow-up telephone survey of 21 violin and viola players with focal dystonia. Eighteen musicians responded to the questionnaire. Information on long-term outcome was available on average 13.8 years after onset of symptoms. Main complaints were playing-related loss of control and involuntary movements affecting the fingering hand in 16 and the bow arm in 5 patients. In 18 patients (86%), signs of abnormal posture could be detected by watching them play their instrument. Treatment attempts included nerve decompression, physical therapy, retraining, and anticholinergic medication. In selected patients, botulinum toxin injections or splint devices were offered. Only 38% of the performing artists were able to maintain their professional careers, among them none with bow arm dystonia. Focal dystonia may affect the fingering hand or bow arm in violin and viola instrumentalists. Treatment benefit is limited and in more than half of the patients, dystonia leads to the end of their musical career. PMID- 14743358 TI - Clinical heterogeneity of neurodegeneration with brain iron accumulation (Hallervorden-Spatz syndrome) and pantothenate kinase-associated neurodegeneration. AB - Hallervorden Spatz syndrome (HSS), also referred to as neurodegeneration with brain iron accumulation (NBIA), is a rare inherited neurodegenerative disorder with childhood, adolescent, or adult onset. Patients with HSS/NBIA have a combination of motor symptoms in the form of dystonia, parkinsonism, choreoathetosis, corticospinal tract involvement, optic atrophy, pigmentary retinopathy, and cognitive impairment. After the recent identification of mutations in the PANK2 gene on chromosome 20p12.3-p13 in some patients with the HSS/NBIA phenotype, the term pantothenate kinase-associated neurodegeneration (PKAN) has been proposed for this group of disorders. To characterize clinically and genetically HSS/NBIA, we reviewed 34 affected individuals from 10 different families, who satisfied the inclusion criteria for NBIA. Relatives of patients who had clinical, magnetic resonance imaging (MRI), or pathological findings of NBIA were included in the study. Four patients were found to have mutations in the pantothenate kinase 2 (PANK2) gene. We compared the clinical features and MRI findings of those with and without PANK2 mutations. The presence of mutation in the PANK2 gene is associated with younger age at onset and a higher frequency of dystonia, dysarthria, intellectual impairment, and gait disturbance. Parkinsonism is seen predominantly in adult-onset patients whereas dystonia seems more frequent in the earlier-onset cases. The phenotypic heterogeneity observed in our patients supports the notion of genetic heterogeneity in the HSS/NBIA syndrome. PMID- 14743360 TI - Parkinsonism in Ontario: comorbidity associated with hospitalization in a large cohort. AB - To study comorbidity in patients with Parkinsonism (PKM), relative hospitalization rates from 1994 to 1999 for 15,304 cases were compared with 30,608 controls. After correction for differential survival, the rates were higher for cases compared to controls for aspiration pneumonia (6.34; 95% confidence interval [CI], 5.23, 7.93), affective psychosis (2.71; 95% CI, 2.13, 3.32), hip fractures (2.56; 95% CI, 2.35, 2.76), other urinary tract disorders including infections (2.5; 95% CI, 2.17, 2.86), septicemia (2.39; 95% CI, 2.02, 2.85) and fluid and electrolyte disorders (2.27; 95% CI, 1.93,2.66). The rates for cardiac, cerebrovascular, and peripheral vascular disease were similar. Preventive measures and aggressive management of these conditions as outpatients may reduce the rates of hospitalization and improve the morbidity and mortality of PKM. PMID- 14743361 TI - One-Hz repetitive transcranial magnetic stimulation of the premotor cortex alters reciprocal inhibition in DYT1 dystonia. AB - Repetitive transcranial magnetic stimulation (rTMS) can produce long-lasting effects not only underneath the site of stimulation, but also at distant connected sites. This study aimed to assess how low frequency rTMS over the premotor area might affect abnormalities in spinal motor function in patients with generalised dystonia associated with the DYT1 gene mutation. We assessed reciprocal inhibition (RI) in a group of 8 manifesting carriers of the DYT1 gene (DYT1) and 10 healthy controls. All subjects then received 20 minutes of 1 Hz rTMS over the premotor area, and RI was assessed again. Before rTMS, the second and third phases of RI were abnormal in DYT1 subjects compared to controls. After 20 minutes of 1 Hz rTMS over the premotor area, a significant increase in inhibition was noted in the third and possibly the first phase of RI in the DYT1 group. No changes in RI were observed in control subjects after rTMS. We have shown for the first time to date that reducing cortical excitability in patients with dystonia using rTMS can produce corresponding changes in abnormal spinal motor output. These findings make a case for further exploring rTMS as a tool to modulate abnormal cortical and spinal excitability in individuals with dystonia and even as a potential form of treatment for dystonic symptoms. PMID- 14743362 TI - Comparison of dementia with Lewy bodies to Alzheimer's disease and Parkinson's disease with dementia. AB - We compared the clinical and neuropsychological pattern of dementia with Lewy bodies (DLB) to Alzheimer's disease (AD) and Parkinson's disease with dementia (PD-d). Sixteen patients clinically diagnosed with DLB were compared with two groups of patients with PD-d (n = 15) and AD (n = 16) matched for level of dementia. Isolated cognitive impairment was the most common form of presentation in AD (93.8%) and DLB (31.3%) groups, while parkinsonism was in 100% of PD-d subjects. Psychoses associated with cognitive impairment at the beginning of the disease were more frequent in DLB patients (31.3%) than in AD (6.3%) and PD-d (0%) groups. There were no significant differences in Unified Parkinson Disease Rating Scale motor-subscale scores between DLB and PD-d patients. DLB and PD-d patients performed significantly worse on attentional functions and better on memory tests than AD. DLB patients also showed lower scores than AD subjects on visual memory, visuoperceptive, and visuoconstructive tests. No significant differences were found between PD-d group and DLB subjects on any neuropsychological test. We were unable to find any differences in cognitive tasks between PD-d and DLB subjects. Clinical features and neuropsychological deficiencies of DLB (attentional, visuoperceptive, and visuoconstructive deficits) and PD (attentional deficits) compared to AD (amnesic syndrome) can contribute to accurate identification of these entities and to the understanding of the neuropathological and neurochemical substrate underlying these diseases. PMID- 14743363 TI - Impaired modulation of the vestibulo-ocular reflex in Huntington's disease. AB - The vestibulo-ocular reflex (VOR) stabilizes gaze during movement, in conjunction with other afferent information: visual, proprioceptive, and somaesthetic. The reflex can either be augmented or suppressed, depending on visual requirements, and undergoes long-term adaptation to compensate for physical changes in the subject. Importantly, over relatively short periods of time, the VOR should function consistently under the same circumstances. This study examines VOR function in patients with Huntington's disease (HD), with a view to investigating cortical influences on the reflex. Horizontal eye movements were recorded in 9 patients with HD and 7 normal subjects, using the scleral search coil technique, in response to high frequency, unpredictable head rotations imposed manually. To establish base VOR function, recordings were made in darkness, without instruction, before and after wearing x2 magnifying lenses for a period of 2 hours to adapt the reflex. Recordings were also made before adaptation, while fixating a stationary visual target (VOR augmentation), and while fixating a target moving with the head (VOR suppression). Although results suggest that the VOR is preserved in HD, with relatively normal gain values and appropriate augmentation and suppression of the reflex with visual input, patients were unable to adapt the VOR to altered visual conditions. This represents a novel finding in HD and suggests that cortical structures compromised in HD exert influences on the long-term adaptation of the VOR. PMID- 14743364 TI - DNA sequence analysis of monoamine oxidase B gene coding and promoter regions in Parkinson's disease cases and unrelated controls. AB - The allele G of the intron 13 G/A polymorphism of the monoamine oxidase B gene (MAO-B) has been associated with Parkinson's disease (PD) in several studies. Apart from a potential direct effect on splicing processes, the association of this intronic polymorphism with PD is due possibly to linkage disequilibrium with other mutations in the coding or promoter regions of the gene. We addressed this latter hypothesis by determining the DNA sequence of the entire MAO-B coding region comprising 15 exons and partial intronic sequences flanking each exon, in 33 cases with idiopathic PD and 38 unrelated controls. The promoter region of MAO B gene up to base -1,369 from ATG (start point of mRNA translation) was also sequenced to identify variants with potential functional effects on gene transcription. In the promoter region, a new polymorphism consisting of a C to T single base change was detected in position -1,114 from ATG, with an allelic frequency of 3.5%, but it was not associated with PD risk. No commonly occurring (>10%) polymorphisms were found in the exons or the intronic sequences flanking the exons, although several rare variants were detected in the coding and promoter regions. PMID- 14743365 TI - Cortical motor activation in akinetic schizophrenic patients: a pilot functional MRI study. AB - Akinesia is associated with supplementary motor area (SMA) dysfunction in Parkinson's disease. We looked for a similar association in patients with schizophrenia. Using functional magnetic resonance imaging (fMRI), we compared motor activation in 6 akinetic neuroleptic-treated schizophrenic patients and 6 normal subjects. Schizophrenic patients had a defective activation in the SMA, left primary sensorimotor cortex, bilateral lateral premotor and inferior parietal cortices, whereas the right primary sensorimotor cortex and a mesial frontal area were hyperactive. SMA was hypoactive in akinetic schizophrenic patients, emphasizing the role of this area in motor slowness. Other abnormal signals likely reflect schizophrenia-related abnormal intracortical connections. PMID- 14743366 TI - Mutational analysis of neurotensin in familial restless legs syndrome. AB - A susceptibility locus for restless legs syndrome (RLS) has been identified recently on chromosome 12q. This region contains several transcribed genes including neurotensin (NTS), which, as an important modulator of the dopaminergic transmission, represents a strong functional and positional candidate in the context of RLS. In this study, NTS was evaluated for mutational analysis. A panel of 19 individuals from 4 families supporting linkage to 12q was investigated using a combined denaturing high-performance liquid chromatography (dHPLC) and direct sequencing method. Analysis of the NTS genomic sequence revealed 2 intronic polymorphisms and 1 variant located in the 5' untranslated region (UTR). None of the observed variants co-segregated with RLS and no disease-associated polymorphisms were detected in any of the analyzed families. Based on these results, it is unlikely that NTS is the gene responsible for RLS in chromosome 12 linked families. PMID- 14743367 TI - Effects of subthalamic nucleus stimulation on characteristics of EMG activity underlying reaction time in Parkinson's disease. AB - We examined the effects of high-frequency deep brain stimulation of the subthalamic nucleus (STN-DBS) on characteristics of electromyographic (EMG) activity of the agonist muscle in 8 patients with Parkinson's disease (PD). Patients were examined during STN-DBS (ON), and 30 minutes after switching off both stimulators (OFF). They were asked to make a ballistic movement in paradigms of simple reaction time (SRT) and choice reaction time (CRT) tasks. Onset of movement (MOVonset) was measured as the latency of the initial displacement from baseline of the signal from an accelerometer attached to the dorsum of the hand. In the associated EMG activity, recorded from wrist extensor muscles, we measured onset latency (EMGonset), size of the first EMG burst (EMGsize), and number of EMG bursts (EMGbursts) counted between EMGonset and task execution. MOVonset and EMGonset were significantly shorter in ON than in OFF conditions in CRT. EMGsize was larger, EMGbursts were reduced, and peak of the acceleration profile was larger in ON compared with OFF conditions in both SRT and CRT. Our results indicate that STN-DBS induces a significant improvement in motor performance of reaction time tasks in PD patients. Such improvement is associated with a change in features of the EMG activity suggesting an increase in the excitability of the motor pathways engaged in ballistic movements. PMID- 14743368 TI - It's a double knock-out! The quaking mouse is a spontaneous deletion of parkin and parkin co-regulated gene (PACRG). AB - Mutations in the parkin gene (PRKN) are the commonest cause of juvenile and early onset parkinsonism. However, the pathogenic mechanism by which loss of parkin protein results in degeneration of dopaminergic neurons remains elusive. Animal models provide a useful tool for the study of development and disease, and the recent production of transgenic fly and mouse parkin deficient models allows investigation of the molecular role of parkin in dopamine regulation and nigrostriatal function. We have identified the mouse mutant Quaking as a spontaneously occurring PRKN knockout. The quaking mutation is a deletion of approximately 1.17 Mb of mouse chromosome 17, resulting in the deletion of the entire promoter and first five coding exons of PRKN In addition, the recently described Parkin Co-Regulated Gene (PACRG) is completely deleted. Homozygous Quaking mice show a complete loss of PRKN and PACRG mRNA and protein. These mice will constitute a useful additional model for studies of the molecular role of parkin and PACRG in neurodegeneration. PMID- 14743369 TI - Michael J. Fox and his Parkinson's disease. AB - Michael J. Fox was a popular and successful film and television comic actor who developed Parkinson's disease at the age of 29 years. His recently published book, Lucky Man, structured around the story of his Parkinson's disease, is an amusing, briskly paced yet introspective memoir that covers the first 40 years of his life. Although quite anecdotal, it contains interesting observations on the preclinical phase of the disorder, evolution of motor fluctuations, and tactics for pharmacological treatment. PMID- 14743370 TI - Adverse effects of dopamine potentiation by long-term treatment with selegiline. AB - A patient with triosephosphate isomerase (TPI) deficiency exhibited worsening of abnormal involuntary movements of the dystonic type and developed psychiatric symptoms while on selegiline. When selegiline was stopped after 9 years of treatment, abnormal involuntary movements improved to pretreatment level and psychiatric behaviour returned to normal. Monoamine oxidase-B platelet activity was low in this patient. PMID- 14743371 TI - Spatial patterns of alpha-synuclein positive glial cytoplasmic inclusions in multiple system atrophy. AB - In cases of multiple system atrophy (MSA), glial cytoplasmic inclusions (GCI) were distributed randomly or present in large diffuse clusters (>1,600 microm in diameter) in most areas studied. These spatial patterns contrast with those reported for filamentous neuronal inclusions in the tauopathies and alpha synucleinopathies. PMID- 14743372 TI - Genetic markers and the majority's right not to know. PMID- 14743373 TI - Re: Yianni et al., Globus pallidus internus deep brain stimulation for dystonic conditions: a prospective audit. PMID- 14743375 TI - Re: Executive cognitive deficits in primary dystonia. PMID- 14743377 TI - Explanatory note. PMID- 14743378 TI - Clarification: Pathogenic role of glial cells in Parkinson's disease. PMID- 14743380 TI - In memory: Dr. Edward Bresnick. PMID- 14743381 TI - Retirement or relearning the joys of the discovery process. PMID- 14743382 TI - Cell cycle checkpoints and their impact on anticancer therapeutic strategies. AB - Cells contain numerous pathways designed to protect them from the genomic instability or toxicity that can result when their DNA is damaged. The p53 tumor suppressor is particularly important for regulating passage through G1 phase of the cell cycle, while other checkpoint regulators are important for arrest in S and G2 phase. Tumor cells often exhibit defects in these checkpoint proteins, which can lead to hypersensitivity; proteins in this class include ataxia telangiectasia mutatated (ATM), Meiotic recanbination 11 (Mre11), Nijmegen breakage syndrome 1 (Nbs 1), breast cancer susceptibility genes 1 and 2 (BRCA1), and (BRCA2). Consequently, tumors should be assessed for these specific defects, and specific therapy prescribed that has high probability of inducing response. Tumors defective in p53 are frequently considered resistant to apoptosis, yet this defect also provides an opportunity for targeted therapy. When their DNA is damaged, p53-defective tumor cells preferentially arrest in S or G2 phase where they are susceptible to checkpoint inhibitors such as caffeine and UCN-01. These inhibitors preferentially abrogate cell cycle arrest in p53-defective cells, driving them through a lethal mitosis. Wild type p53 can prevent abrogation of arrest by elevating levels of p21(waf1) and decreasing levels of cyclins A and B. During tumorigenesis, tumor cells frequently loose checkpoint controls and this facilitates the development of the tumor. However, these defects also represent an Achilles heel that can be targeted to improve current therapeutic strategies. PMID- 14743383 TI - Modulation of phosphatidylinositol-3-kinase/protein kinase B- and mitogen activated protein kinase-pathways by tea polyphenols in human prostate cancer cells. AB - We have earlier shown that oral infusion of a polyphenolic fraction isolated from green tea, at a human achievable dose (equivalent to six cups of green tea per day), significantly inhibits prostate cancer (PCA) development and metastasis in transgenic adenocarcinoma of mouse prostate (TRAMP) model that closely mimics progressive form of human prostatic disease (Gupta et al. [2001]: Proc. Natl. Acad. Sci. U.S.A. 98:10350-10355.). A complete understanding of the mechanism(s) and molecular targets of PCA chemopreventive effects of tea polyphenols may be useful in developing novel approaches for its prevention. In this study, we employed two distinct human PCA cell lines viz. DU145 (androgen-unresponsive prostate carcinoma cells) and LNCaP (androgen-responsive prostate carcinoma cells) and, employing immunoblot analysis, we evaluated the effect of epigallocatechin-3-gallate (EGCG), the major polyphenol present in green tea and theaflavins (TF), the major polyphenol present in black tea on phosphatidylinositol-3-kinase (PI3K)/protein kinase B (PKB) and mitogen-activated protein kinase (MAPK) pathways. Both EGCG and TF treatment were found to (i) decrease the levels of PI3K and phospho-Akt and (ii) increase Erk1/2 in both DU145 and LNCaP cells. Our data showing the inhibition of the constitutive levels of PI3K and the phosphorylation of Akt could be important because the treatment approaches should be aimed at the inhibition of the constitutive levels of PI3K and Akt. Our data also suggest that Erk1/2 could be involved in the anti-cancer effects of EGCG and TF. Taken together, our study, for the first time demonstrated the modulation of the constitutive activation of PI3K/Akt and Erk1/2 pathways by EGCG as well as TF. We suggest that detailed studies in appropriate tumor model system are needed to establish the relevance of the cell culture work to in vivo models. PMID- 14743384 TI - Hyperplasia, partial hepatectomy, and the carcinogenicity of aflatoxin B1. AB - Generalized cellular hyperplasia has long been associated as a factor in the causation of liver cancer. Parenchymal cell hyperplasia resulting from hepatotoxins, viruses, parasites, or malnutrition is exceedingly variable as to when it occurs, its extent, and its duration. Partial hepatectomy has been used as an experimental tool precisely because the timing and extent of hyperplasia can be known and controlled. With regards to aflatoxin B1 (AFB1) carcinogenesis, partial hepatectomy has produced variable results. An explanation appears to reside in the hepatotoxic properties of AFB1 that enhance the early stages of carcinogenesis. PMID- 14743385 TI - The biological effects of N3-methyladenine. AB - The targeting of damage to DNA remains an attractive strategy to kill tumor cells. One of the serious side effects of alkylating agents is that they create both toxic (desired) and mutagenic (undesired) lesions. The result is that patients successfully treated for a primary cancer are at significant risk to develop cancer related to their therapy. To address this issue we have prepared agents that selectively methylate DNA at the N3-position of adenine. The presence of this lesion in DNA is thought to halt DNA polymerase, and this then initiates a cascade of events including cell death. The toxicity and mutagenicity of the compound, Me-lex, used to generate N3-methyladenine is discussed in bacterial, yeast, and mammalian systems. Mechanisms are proposed to explain the biological activities of N3-methyladenine. PMID- 14743386 TI - Identification of molecular targets for dietary energy restriction prevention of skin carcinogenesis: an idea cultivated by Edward Bresnick. AB - Dietary energy restriction (DER) has long been known to strikingly inhibit carcinogenesis in many animal models. The animal data has been corroborated by recent and ongoing epidemiological studies demonstrating the importance of energy balance, physical exercise and obesity in human cancer. Dr. Edward Bresnick provided key insights into this important area of research and pivotal direction for the author's research while he served as Director of the Eppley Institute for Research in Cancer, Omaha, NE. These insights moved this research toward demonstrating that DER reduced the expression of key protein kinase C isoforms in mouse skin. More recent studies have uncovered downstream events that are inhibited by DER including blockage of tumor promoter activation of Raf-1, ERK 1,2 and AP-1 expression. Parallel studies have demonstrated the DER inhibition of these key cellular signaling events in mouse skin carcinogenesis are dependent upon an intact adrenal gland because adrenalectomized mice fed DER diet did not have reduced tumor burden or inhibited signaling and blocked AP-1 activation as was observed in DER mice with intact adrenal glands. In addition, the DER inhibition of tumorigenesis and AP-1 signaling was restored in adrenalectomized mice that were given corticosterone in the drinking water. This showed that in mice in the chemical carcinogenesis protocol glucocorticoid hormone plays a major role in mediating DER prevention of cancer. Studies are ongoing to further assess the mechanism of DER modulation of skin cancer by assessing impacts on transcriptional regulation and expression of genes that are critical in skin carcinogenesis. PMID- 14743387 TI - Breast cancer cells induce osteoblast apoptosis: a possible contributor to bone degradation. AB - Breast cancer cells exhibit a predilection for metastasis to bone. There, the metastases usually bring about bone loss with accompanying pain and loss of function. One way that breast cancer cells disrupt the normal pattern of bone remodeling is by activating osteoclasts, the bone degrading cells. Nevertheless, targeting the osteoclasts does not cure the disease or result in bone repair. These observations indicate that osteoblast function also may be compromised. The objective of this study was to investigate the interaction of metastatic breast cancer cells with osteoblasts. Human metastatic breast cancer cells, MDA-MB-435 or MDA-MB-231, or their conditioned media were co-cultured with a human osteoblast line hFOB1.19. The breast cancer cells caused an increase in the prevalence of apoptotic osteoblasts. Apoptotic osteoblasts detected by the TUNEL assay or by caspase activity increased approximately two to fivefold. This increase was not seen with non-metastatic MDA-MB-468 cells. In an investigation of the mechanism, it was determined that the hFOB1.19 cells expressed fas and that fas was functional. Likewise the hFOB1.19 cells were susceptible to TNF alpha, but this cytokine was not detected in the conditioned medium of the breast cancer cells. This study indicates that osteoblasts are the target of breast cancer cell-induced apoptosis, but fas/fas-ligand and TNF-alpha, two common initiators of cell death, are probably not involved in this aspect of the metastases/bone cell axis. There are several mechanisms that remain to be explored in order to determine how breast cancer cells bring about osteoblast apoptosis. Even though the specific initiator of apoptosis remains to be identified, the results of this study suggest that the mechanism is likely to be novel. PMID- 14743388 TI - Tailoring cancer chemoprevention regimens to the individual. AB - The present article, which is a tribute to the memory of Dr. Edward Bresnick, emphasizes the importance of environmental and life-style factors for cancer causation in the human population and points out approaches to cancer prevention. These approaches include vaccinations for the prevention of cancers that are caused by infectious agents as well as the use of cancer chemopreventive agents. The use of tamoxifen and letrozole to prevent breast cancer, finasteride to prevent prostate cancer, sunscreens or topical applications of 5-fluorouracil to prevent sunlight-induced skin cancer, and aspirin or calcium to prevent colon cancer are a few examples of cancer chemoprevention in high risk individuals and in the general population. An underdeveloped area of cancer chemoprevention is the use of combinations of agents that work by different mechanisms. It was pointed out that animal studies indicate that many cancer chemopreventive agents inhibit carcinogenesis under one set of experimental conditions but enhance carcinogenesis under another set of experimental conditions. These observations suggest that tailoring the chemopreventive regimen to the individual or to groups of individuals living under different environmental conditions or with different mechanisms of carcinogenesis may be an important aspect of cancer chemoprevention in human populations. How to tailor cancer chemoprevention regimens to the individual is an important challenge for the future. PMID- 14743389 TI - Nuclear microenvironments support assembly and organization of the transcriptional regulatory machinery for cell proliferation and differentiation. AB - The temporal and spatial organization of transcriptional regulatory machinery provides microenvironments within the nucleus where threshold concentrations of genes and cognate factors facilitate functional interactions. Conventional biochemical, molecular, and in vivo genetic approaches, together with high throughput genomic and proteomic analysis are rapidly expanding our database of regulatory macromolecules and signaling pathways that are requisite for control of genes that govern proliferation and differentiation. There is accruing insight into the architectural organization of regulatory machinery for gene expression that suggests signatures for biological control. Localized scaffolding of regulatory macromolecules at strategic promoter sites and focal compartmentalization of genes, transcripts, and regulatory factors within intranuclear microenvironments provides an infrastructure for combinatorial control of transcription that is operative within the three dimensional context of nuclear architecture. PMID- 14743390 TI - Gene array identification of osteoclast genes: differential inhibition of osteoclastogenesis by cyclosporin A and granulocyte macrophage colony stimulating factor. AB - Treatment of adherent peripheral blood mononuclear cells (PBMCs) with macrophage colony stimulating factor (M-CSF) and receptor activator of NF-kappaB ligand (RANKL) stimulates the formation of multinucleate osteoclast-like cells. Treatment with M-CSF alone results in the formation of macrophage-like cells. Through the use of Atlas human cDNA expression arrays, genes regulated by RANKL were identified. Genes include numerous cytokines and cytokine receptors (RANTES and CSF2R proportional, variant ), transcription factors (nuclear factor of activated T-cells cytoplasmic 1 (NFATc1) and GA binding protein transcription factor alpha (GABPalpha)), and ribosomal proteins (60S L17 and 40S S20). Real time PCR analysis showed significant correlation (R2 of 0.98 P < 0.01) with array data for all genes tested. Time courses showed differential activation patterns of transcription factors with early induction of FUSE binding protein 1 (FBP) and c-Jun, and later steady upregulation of NFATc1 and GABP by RANKL. Treatment with cyclosporin A, a known NFATc1 inhibitor, resulted in a blockade of osteoclast formation. The mononuclear cells resulting from high cyclosporin treatment (1,000 ng/ml) were cathepsin K (CTSK) and tartrate-resistant acid phosphatase (TRAP) positive but expression of calcitonin receptor (CTR) was downregulated by more than 30-fold. Constant exposure of M-CSF- and RANKL-treated cells to GM-CSF resulted in inhibition of osteoclast formation and the downregulation of CTSK and TRAP implicating the upregulation of CSF2R in a possible feedback inhibition of osteoclastogenesis. PMID- 14743391 TI - Platelet-derived growth factor-BB phosphorylates heat shock protein 27 in cardiac myocytes. AB - It is recognized that heat shock protein 27 (HSP27) is highly expressed in heart. In the present study, we investigated whether platelet-derived growth factor (PDGF) phosphorylates HSP27 in mouse myocytes, and the mechanism underlying the HSP27 phosphorylation. Administration of PDGF-BB induced the phosphorylation of HSP27 at Ser-15 and -85 in mouse cardiac muscle in vivo. In primary cultured myocytes, PDGF-BB time dependently phosphorylated HSP27 at Ser-15 and -85. PDGF BB stimulated the phosphorylation of p44/p42 mitogen-activated protein (MAP) kinase, p38 MAP kinase, and stress-activated protein kinase/c-Jun N-terminal kinase (SAPK/JNK) among the MAP kinase superfamily. SB203580, a specific inhibitor of p38 MAP kinase, reduced the PDGF-BB-stimulated phosphorylation of HSP27 at both Ser-15 and -85, and phosphorylation of p38 MAP kinase. However, PD98059, a specific inhibitor of MEK, or SP600125, a specific inhibitor of SAPK/JNK, failed to affect the HSP27 phosphorylation. These results strongly suggest that PDGF-BB phosphorylates HSP27 at Ser-15 and -85 via p38 MAP kinase in cardiac myocytes. PMID- 14743392 TI - Cleavage of p130Cas in anoikis. AB - p130Cas is a multifunctional signaling adaptor protein. It integrates and relays signals generated from a variety of extracellular stimuli and regulates a number of cellular activities including cell death. In this study, we analyzed the regulation and function of p130Cas in anoikis, a type of apoptosis caused by disruption of cell-matrix interactions. We found that p130Cas was specifically cleaved during anoikis in anoikis-sensitive epithelial cells, but not in anoikis resistant tumor cells. There is a close correlation between p130Cas cleavage and anoikis. Furthermore, we found that the cleavage of p130Cas, as well as another focal adhesion component FAK, is different from that of caspase substrate PARP and spectrin. Although caspases and calpain were found to be involved in the cleavage of p130Cas, there appear to be other unidentified proteases that are mainly responsible for the cleavage of p130Cas, particularly at the early stage of anoikis. Overexpression of the p130Cas cleavage product induced apoptosis. Taken together, these data suggest that there are novel proteases involved in the cleavage of p130Cas during anoikis, which may be functionally involved in the onset of anoikis. p130Cas may have a dual role in the regulation of anoikis. On one hand, it mediates a survival signal from cell-matrix interactions when cells are attached to the extracellular matrix. On the other hand, it participates in executing cell death when cell-matrix interactions are disrupted. These observations provide new insights into the understanding of the function of p130Cas and the molecular mechanism of anoikis. PMID- 14743393 TI - PPARgamma1 synthesis and adipogenesis in C3H10T1/2 cells depends on S-phase progression, but does not require mitotic clonal expansion. AB - Adipogenesis is typically stimulated in mouse embryo fibroblast (MEF) lines by a standard hormonal combination of insulin (I), dexamethasone (D), and methylisobutylxanthine (M), administered with a fresh serum renewal. In C3H10T1/2 (10T1/2) cells, peroxisome proliferator-activated receptor gamma1 (PPARgamma1) expression, an early phase key adipogenic regulator, is optimal after 36 h of IDM stimulation. Although previous studies provide evidence that mitotic clonal expansion of 3T3-L1 cells is essential for adipogenesis, we show, here, that 10T1/2 cells do not require mitotic clonal expansion, but depend on cell cycle progression through S-phase to commit to adipocyte differentiation. Exclusion of two major mitogenic stimuli (DM without insulin and fresh serum renewal) from standard IDM protocol removed mitotic clonal expansion, but sustained equivalent PPARgamma1 synthesis and lipogenesis. Different S-phase inhibitors (aphidicolin, hydroxyurea, l-mimosine, and roscovitin) each arrested cells in S-phase, under hormonal stimulation, and completely blocked PPARgamma1 synthesis and lipogenesis. However, G2/M inhibitors effected G2/M accumulation of IDM stimulated cells and prevented mitosis, but fully sustained PPARgamma1 synthesis and lipogenesis. DM stimulation with or without fresh serum renewal elevated DNA synthesis in a proportion of cells (measured by BrdU labeling) and accumulation of cell cycle progression in G2/M-phase without complete mitosis. By contrast, standard IDM treatments with fresh serum renewal caused elevated DNA synthesis and mitotic clonal expansion while achieved equivalent level of adipogenesis. At most, one-half of the 10T1/2 mixed cell population differentiated to mature adipocytes, even when clonally isolated. PPARgamma was exclusively expressed in the cells that contained lipid droplets. IDM stimulated comparable PPARgamma1 synthesis and lipogenesis in isolated cells at low cell density (LD) culture, but in about half of the cells and with sensitivity to G1/S, but not G2/M inhibitors. Importantly, growth arrest occurred in all differentiating cells, while continuous mitotic clonal expansion occurred in non-differentiating cells. Irrespective of confluence level, 10T1/2 cells differentiate after progression through S-phase, where adipogenic commitment induced by IDM stimulation is a prerequisite for PPARgamma synthesis and subsequent adipocyte differentiation. PMID- 14743394 TI - Differentially up-regulated genes in proliferating porcine neonatal pancreas cells caused by epidermal growth factor. AB - Pancreatic duct cells are considered to be a major source for beta-cell regeneration or neogenesis. Although epidermal growth factor (EGF) is a well known important growth factor for pancreas development, the control of pancreatic duct cell growth and differentiation by EGF is poorly understood. In this study, we focused on identifying the genes that were differentially up-regulated in response to EGF stimulation using monolayer cultured porcine neonatal pancreas cells. Cells were obtained from 1 to 3 day old pigs, dispersed and cultured for 8 days. Monolayer cultured porcine pancreas cells were comprised of duct cells and some endocrine and mesenchymal cells (75.2 +/- 15.1, 19.6 +/- 4.9, and 9.5 +/- 3.1%, respectively). After 16 h in serum free media, cells were treated with 100 microg/L EGF for 24 h. Differentially expressed genes were screened by subtractive hybridization. (3)H-thymidine uptake was significantly increased by EGF with time (untreated vs. 24 h treated, untreated vs. 48 h treated: 305.5 +/- 3.5 cpm vs. 380.3 +/- 17.3 cpm (P < 0.05), 309.2 +/- 4.51 vs. 929 +/- 9.19 cpm, (P < 0.005), respectively). Three hundred and fifty cDNA clones were obtained by subtractive hybridization and the inserts were confirmed in 161 colonies and then sequenced. Finally, we found increased mRNA expression of five unknown and five known genes, including cytochrome c oxidase subunit I (COI), cyclooxygenase-2 (COX-2), matrix metalloproteinase-13 (MMP-13), Wiskott-Aldrich syndrome protein interacting protein (WASPIP), and hyaluronan synthase-2 (HAS-2). We confirmed the up-regulation of these genes by Northern blot and semi-quantitative RT-PCR at various time points. The present findings opened new targets for the research on the mechanisms of pancreatic duct cell proliferation by EGF. PMID- 14743395 TI - Colon cancer specific nuclear matrix protein alterations in human colonic adenomatous polyps. AB - Most colon cancers arise within preexisting adenomatous polyps or adenomas. The slow evolution from the non-invasive premalignant lesion, the adenomatous polyp, to invasive cancer supports a strategy of early detection. Recently, we identified unique nuclear matrix proteins (NMPs) specific for colon cancer (CC2, CC3, CC4, CC5). Most of the NMPs identified are common to all cell types, but several identified NMPs are tissue and cell line specific. The objective of this study is to describe and characterize the NMP profile of premalignant adenomatous colon polyps. Specifically when in the adenoma-carcinoma sequence four specific colon cancer NMPs, previously described, appear. Using two-dimensional (2-D) gel analysis 20 colon polyps (one juvenile polyp, six tubular adenoma (TA), seven tubulovillous adenoma (TVA), six TVA with focal high-grade dysplasia (HGD), were analyzed for the presence of four (CC2, CC3, CC4, CC5) specific NMPs. CC2 was not seen in any of the premalignant polyps. CC5 was present in only two premalignant TVA with HGD and in one TA. CC3 and CC4 were present in most adenomas. None of the NMPs were seen in the juvenile polyp, which is not considered to be a precursor of colon cancer. CC2 and CC5 are NMPs expressed at the junction of an advanced adenoma and invasive colorectal cancer. CC3 and CC4 are expressed earlier in the evolution of adenomatous polyps. Development of an assay to these proteins may serve as a new method for early detection of colorectal cancer. PMID- 14743396 TI - Regulatory factor X2 (RFX2) binds to the H1t/TE1 promoter element and activates transcription of the testis-specific histone H1t gene. AB - Transcription of the mammalian testis-specific linker histone H1t gene occurs only in pachytene primary spermatocytes during spermatogenesis. Studies of the wild type (Wt) and mutant H1t promoters in transgenic mice show that transcription of the H1t gene is dependent upon the TE promoter element. We purified an 85 kDa protein from rat testis nuclear extracts using the TE1 subelement as an affinity chromatography probe and analysis revealed that the protein was RFX2. The TE1 element is essentially an X-box DNA consensus element and regulatory factor X (RFX) binds specifically to this element. Polyclonal antibodies directed against RFX2 supershift the low mobility testis nuclear protein complex formed in electrophoretic mobility shift assays (EMSA). RFX2 derived from primary spermatocytes, where the transcription factor is relatively abundant, binds with high affinity to the TE1 element. Coexpression of RFX2 together with an H1t promoter/reporter vector activates the H1t promoter in a cultured GC-2spd germinal cell line, but mutation of either the TE1 subelement or the TE2 subelements represses activity. These observations lead us to conclude that the TE1 and TE2 subelements of the testis-specific histone H1t promoter are targets of the transcription factor RFX2 and that this factor plays a key role in activating transcription of the H1t gene in primary spermatocytes. Published 2003 Wiley-Liss, Inc. PMID- 14743397 TI - Mediating of caspase-independent apoptosis by cadmium through the mitochondria ROS pathway in MRC-5 fibroblasts. AB - Cadmium (Cd) is an environmental pollutant of global concern with a 10-30-year biological half-life in humans. Accumulating evidence suggests that the lung is one of the major target organs of inhaled Cd compounds. Our previous report demonstrated that 100 microM Cd induces MRC-5 cells, normal human lung fibroblasts, to undergo caspase-independent apoptosis mediated by mitochondrial membrane depolarization and translocation of apoptosis-inducing factor (AIF) from mitochondria into the nucleus. Here, using benzyloxycarbonyl-Val-Ala-Asp-(ome) fluoromethyl ketone (Z-VAD.fmk) as a tool, we further demonstrated that Cd could induce caspase-independent apoptosis at concentrations varied from 25 to 150 microM, which was modulated by reactive oxygen species (ROS) scavengers, such as N-acetylcysteine (NAC), mannitol, and tiron, indicating that ROS play a crucial role in the apoptogenic activity of Cd. Consistent with this notion, the intracellular hydrogen peroxide (H2O2) was 2.9-fold elevated after 3 h of Cd treatment and diminished rapidly within 1 h as detected by flow cytometry with 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA) staining. Using inhibitors of the mitochondrial electron transport chain (ETC) (oligomycin A and rotenone for complex I and V, respectively) and mitochondrial permeability transition pore (MPTP) (cyclosporin A and aristolochic acid), we coincidently found the ROS production, mitochondrial membrane depolarization, and apoptotic content were almost completely or partially abolished. As revealed by confocal microscopy staining with chloromethyl-X-rosamine (CMXRos) and an anti-AIF antibody, the collapse of mitochondrial membrane potential induced by Cd (3 h-treatment) was a prelude to the translocation of caspase-independent pro-apoptotic factor, AIF, into the nucleus (after 4 h of Cd treatment). In summary, this study demonstrated that, in MRC-5 fibroblasts, Cd induced caspase-independent apoptosis through a mitochondria-ROS pathway. More importantly, we provide several lines of evidence supporting a role of mitochondrial ETC and MPTP in the regulation of caspase independent cell death triggered by Cd. PMID- 14743399 TI - Transcriptional regulation of TNF family receptors and Bcl-2 family by chemotherapeutic agents in murine CT26 cells. AB - Various chemotherapeutic agents have been shown to sensitize cancer cells to members of the tumor necrosis factor (TNF) family. However, it is unclear whether sensitization by chemotherapeutic agents involves the transcriptional regulation of apoptosis-related genes. In this study, we investigated mRNA regulation of TNF family receptors and Bcl-2 family members after treating the murine colon cancer cell line, CT26, with various apoptosis inducers. We found that treatment with cycloheximide, a protein synthesis inhibitor, remarkably increased CD40 mRNA levels by semi-quantitative RT-PCR. Other protein synthesis inhibitors, such as anisomycin and emetine, also enhanced CD40 mRNA expression, which was significantly blocked by a NF-kappaB antagonist and a p38 MAP kinase antagonist. After treatment with cycloheximide, and further cultivation in fresh medium, CD40 protein levels were found to increase by flow cytometry. Additionally, we found that cycloheximide treatment appeared to downregulate the Bcl-xL mRNA level but not the Bax mRNA level by RNase protection assay. Because the upregulation of CD40 mRNA and the downregulation of Bcl-xL correlated with CT26 cell death, our results suggest that chemotherapeutic agents, including cycloheximide, may exert their synergistic effects on the TNF family treatment of cancer cells by regulating the mRNA levels of apoptosis-related genes. PMID- 14743398 TI - Angiostatin-induced inhibition of endothelial cell proliferation/apoptosis is associated with the down-regulation of cell cycle regulatory protein cdk5. AB - Endothelial cells (ECs) are quiescent in normal blood vessels, but undergo rapid bursts of proliferation after vascular injury, hypoxia or induced by powerful angiogenic cytokines like fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF). Deregulated proliferation of ECs facilitates angiogenic processes and promotes tumor growth. In dividing cells, cell cycle associated protein kinases, which are referred as cyclin-dependent kinases (cdks), regulate proliferation, differentiation, senescence, and apoptosis. Cyclin-dependent kinase-5 (cdk5) is expressed in neuronal cells and plays an important role in neurite outgrowth, of neuronal migration and neurogenesis, its functions in non-neuronal cells are unclear. Here, we show for the first time that the cdk5 is expressed at high levels in proliferating bovine aortic endothelial (BAE) cells, by contrast insignificant low levels of cdk5 expression in quiescent BAE cells. In addition, bFGF up-regulates cdk5 expression in a dose dependent fashion. Interestingly, temporal expression data suggests that cdk5 expression is very low between 24-48 h, but high level of cdk5 expression was detected during 60-72 h. This later time corresponds to the time of completion of one cell cycle (doubling of cell population) of BAE cell culture. Angiostatin (AS), a powerful inhibitor of angiogenesis inhibits ECs proliferation in dose dependent manner with concomitant down-regulation of cdk5 expression. The role of cdk5 in ECs, proliferation and apoptosis was confirmed by selective inhibition of cdk5 expression by the purine derivative roscovitine, which inhibits bFGF stimulated BAE cells proliferation and induces apoptosis in dose-specific manner. By contrast, the roscovitine analog olomoucine, which is a specific inhibitor of cdk4, but not of cdk5 failed to affect ECs proliferation and apoptosis. These data suggest for the first time that neuron specific protein cdk5 may have significant role in the regulation of ECs proliferation, apoptosis, and angiogenesis and extends beyond its role in neurogenesis. PMID- 14743400 TI - Constitutive and inducible expression of cytochromes P4501A (CYP1A1 and CYP1A2) in normal prostate and prostate cancer cells. AB - Constitutive and benzo[a]pyrene (B[a]P) inducible expression of CYP1A1 and CYP1A2 in prostate cancer and normal prostate epithelial cells were examined by immunoblotting. Androgen independent prostate cancer cell lines DU145 and PC3 have constitutive expression of CYP1A and CYP1A1 and CYP1A2, respectively. Four micromolar B[a]P did not appear to induce CYP1A1 or CYP1A2 expression in DU145 or PC3 cells. The androgen dependent prostate cancer cell line, LnCap, also has constitutive expression of CYP1A1 and CYP1A2. However, both CYP1A1 and CYP1A2 are induced by treatment of LnCap cells with 4 microM B[a]P. Untreated normal prostate and primary prostate tumor cells have no detectable CYP1A1 expression. Treatment with 4 microM B[a]P induced CYP1A1 expression in both normal and primary tumor prostate cells. Constitutive CYP1A2 expression was detected in normal prostate cells with little or no induction by exposure to 4 microM B[a]P. Primary prostate tumor cells did not show constitutive expression of CYP1A2. However, CYP1A2 was induced by 4 microM B[a]P in primary prostate tumor cells. These observations indicate that hormonal and cancer specific factors affect the expression and induction of the phase I metabolic enzymes, CYP1A1 and CYP1A2 in prostate cells. These observations may be related to the potential smoking-linked higher risk of prostate cancer development and morbidity of prostate cancer patients who smoke. PMID- 14743404 TI - Trigonal prismatic or not trigonal prismatic? On the mechanisms of oxygen-atom transfer in molybdopterin-based enzymes. PMID- 14743405 TI - New catalytic approaches in the stereoselective Friedel-Crafts alkylation reaction. AB - After more than 125 years, the Friedel-Crafts alkylation is still one of the most studied and most utilized reactions in organic synthesis. What is the secret of this astonishing success? Perhaps the great versatility in scope and applicability continues to justify its crucial role in the synthesis of more and more complex molecules. However, it has taken more than a century for asymmetric catalytic versions of this reaction to be developed and subsequently extended to a range of aromatic compounds and alkylating agents. Herein we review recent developments in the design and use of catalytic and stereoselective strategies for the alkylation of aromatic systems and synthesis of a wide range of polyfunctionalized enantiomerically enriched compounds. PMID- 14743406 TI - Emerging supramolecular chemistry of gases. AB - Molecular recognition of gases is an emerging area of chemistry. Supramolecular chemistry helps us to understand how gases interact with biological molecules and offers delicate insights into the mechanisms of their physiological activity. Principles of molecular recognition have been used for gas sensing, and have provided fundamental knowledge about the structure and dynamics of receptor analyte complexes, and novel materials for gas sensing and storage have been developed. Supramolecular chemistry is also enabling us to learn how to transform gases into synthetically useful reagents. The rational design of novel catalysts for gas conversion and, more recently, encapsulation complexes with gases open novel directions in preparative synthetic chemistry. PMID- 14743407 TI - Anion encapsulation by neutral supramolecular assemblies of cyclic CuII complexes: a series of five polymerization isomers, [(cis-CuII(mu-OH)(mu-pz))n], n=6, 8, 9, 12, and 14. PMID- 14743408 TI - The origin of the magnetic moments in compressed crystals of polymeric C60. PMID- 14743409 TI - Noncovalent ligand strands for transition-metal helicates: the straightforward and stereoselective self-assembly of dinuclear double-stranded helicates using hydrogen bonding. PMID- 14743410 TI - Sigma-bond stretching: a static approach for a dynamic process. PMID- 14743411 TI - Anti-Markovnikov hydrofunctionalization of olefins mediated by rhodium-porphyrin complexes. PMID- 14743412 TI - Imines in Stille-type cross-coupling reactions: a multicomponent synthesis of alpha-substituted amides. PMID- 14743414 TI - Colloidal carbon spheres and their core/shell structures with noble-metal nanoparticles. PMID- 14743413 TI - How single and bifurcated hydrogen bonds influence proton-migration rate constants, redox, and electronic properties of phenoxyl radicals. PMID- 14743415 TI - Automated solid-phase synthesis of protected tumor-associated antigen and blood group determinant oligosaccharides. PMID- 14743416 TI - The shape of neutral valine. PMID- 14743417 TI - Asymmetric induction in ruthenium-catalyzed [2+2] cycloadditions between bicyclic alkenes and a chiral acetylenic acyl sultam. PMID- 14743419 TI - A tandem epoxidation/stereoselective intramolecular [4+3] cycloaddition reaction involving nitrogen-stabilized oxyallyl cations derived from chiral allenamides. PMID- 14743418 TI - Glycosyltransferase-catalyzed synthesis of thiooligosaccharides. PMID- 14743420 TI - Solid-state reshaping of crystals: flash increase in porosity of zirconium phosphate-hypophosphite that contains polyethylenoxa diphosphonate pillars. PMID- 14743421 TI - Combinatorial synthesis of functionalized 1,3-thiazine libraries using a combined polymer-supported reagent/catch-and-release strategy. PMID- 14743422 TI - Environmental fluctuations facilitate electron-hole transfer from guanine to adenine in DNA pi stacks. PMID- 14743423 TI - Copper(I) cyanide: a simple compound with a complicated structure and surprising room-temperature reactivity. PMID- 14743424 TI - Selenocyclizations: control by coordination and by the counterion. PMID- 14743425 TI - A porphyrin dye with monoexponential fluorescence intensity and anisotropy decay behavior in spherical micelles. PMID- 14743426 TI - Synthesis and structure of a 1,3-diphosphacyclobutadienediide: an aniomesolytic fragmentation of a 1,3-diphosphetane-2,4-diyl in solution. PMID- 14743428 TI - Proteomic analysis of protein changes developing in rat hippocampus after chronic antidepressant treatment: Implications for depressive disorders and future therapies. AB - It is recognized that monoamine reuptake inhibitors (MARIs) exert beneficial effects in the treatment of major depression and general anxiety disorder. The aim of this study was to identify proteins regulated by this class of antidepressant using a proteome differential profiling approach. Either venlafaxine or fluoxetine was administered systemically to adult rats for 2 weeks, and protein patterns from rat hippocampal cytosolic extracts were compared by two-dimensional gel electrophoresis. Silver-stained protein spots displaying differential expression were identified by mass spectrometry. Thirty-three protein spots were modulated by both drug treatments compared to controls. The classification of several proteins that were sorted by function suggested convergent pathway activities for both MARIs at the post-receptor level. These included proteins associated with neurogenesis (insulin like growth factor 1 (IGF 1), glia maturation factor [GMF]-beta), outgrowth/maintenance of neuronal processes (hippocampal cholinergic neurostimulating peptide [HCNP], PCTAIRE-3), and with neural regeneration/axonal guidance collapsin response mediator protein (CRMP-2) systems. Other modulated proteins indicated an increase in neuronal vesicular cell trafficking and synaptic plasticity (Ras-related protein 4a (Rab4a), Ras-related protein 1b (Rab1b), heat shock protein 10 [HSP10]), as well as neurosteroidogenic (hydroxysteroid sulfotransferase A) and possible anti apoptotic (dimethylargininase-1 L-N,N-dimethylarginine dimethylaminohydrolase-1 [DDAH-1], pyruvate dehydrogenase-E1 [PDH-E1], antioxidant protein-2 [AOP-2]) pathway-mediated regulatory events. Parallel studies to investigate further the effects of venlafaxine and fluoxetine on adult hippocampal neurogenesis in vivo by quantitative bromodeoxyuridine immunolabeling revealed a significant drug induced increase in the proliferation rate and long-term survivability of progenitor stem cells located in the subgranular zone. These data suggest that MARIs share wide-ranging proteome changes within the hippocampal formation, beyond 5-HT/NE neurotransmission. This may reflect long-term functional adaptations required for antidepressant activity. PMID- 14743430 TI - Divalent cations modulate the activity of metabotropic glutamate receptors. AB - Metabotropic glutamate receptors (mGluRs) and calcium receptors (CaR) are closely related G protein-coupled receptors (GPCRs). The similar structural and functional properties of mGluRs and CaRs include conserved amino acid residues involved in glutamate binding in mGluRs and Ca2+ binding in the CaR. Furthermore, recent findings have demonstrated that mGluRs can respond to high extracellular Ca2+ (Ca2+(o)) whereas CaR activity is potentiated by L-amino acids. We show that both mGluR1 and mGluR2 are activated by Ca2+(o) in the absence of glutamate in the extracellular media. This activation by Ca2+(o) is antagonized by Mg2+(o). Unlike the CaR, in which the intracellular carboxyl tail has been reported to be involved in Ca2+(o)-dependent activity, the carboxyl tail of mGluRs does not seem to play a role in mediating Ca2+(o) actions. On the other hand, we find that preservation of disulfide bonds in the N-terminal extracellular domain of mGluRs is essential for stimulation by Ca2+(o) as well as glutamate. Because the mGluR1 EC50 for Ca2+(o) is within the physiologic range of Ca2+ in the synaptic cleft, mGluR function is likely regulated by changes in divalent cations caused by synaptic activity under normal or pathologic conditions. PMID- 14743429 TI - Identification of a protein that interacts with the golli-myelin basic protein and with nuclear LIM interactor in the nervous system. AB - The myelin basic protein (MBP) gene encodes the classic MBPs and the golli proteins, which are related structurally to the MBPs but are not components of the myelin sheath. A yeast two-hybrid approach was used to identify molecular partners that interact with the golli proteins. A mouse cDNA was cloned that encoded a protein of 261 amino acids and called golli-interacting protein (GIP). Database analysis revealed that GIP was the murine homolog of human nuclear LIM interactor-interacting factor (NLI-IF), a nuclear protein whose function is just beginning to be understood. It is a member of a broad family of molecules, found in species ranging from yeast to human, that contain a common domain of approximately 100 amino acids. Immunocytochemical and Northern blot analyses showed co-expression of GIP and golli in several neural cell lines. GIP and golli also showed a similar developmental pattern of mRNA expression in brain, and immunohistochemical staining of GIP and golli showed co-expression in several neuronal populations and in oligodendrocytes in the mouse brain. GIP was localized predominantly in nuclei. GIP co-immunoprecipitated with golli in several in vitro assays as well as from PC12 cells under physiologic conditions. GIP was the first member of this family shown to interact with nuclear LIM interactor (NLI). NLI co-immunoprecipitated with GIP and golli from lysates of N19 cells transfected with NLI, further confirming an interaction between golli, GIP, and NLI. The ability of GIP to interact with both golli and NLI, and the nuclear co-localization of GIP and golli in many cells, indicates a role for the golli products of the MBP gene in NLI- associated regulation of gene expression. PMID- 14743431 TI - Differential subcellular distribution of Ca2+/calmodulin-dependent protein kinase II isoforms in the striatum and NG108-15 cells. AB - Four subunits of Ca2+/calmodulin-dependent protein kinase II (CaM KII) have several isoforms, which differ in the variable domain. We previously reported that all subunits were highly expressed in rat striatal neurons. To examine intracellular distributions of CaM KII subunits in the rat striatal neurons, we performed immunoblot analysis with antibodies specific to each subunit in cell extracts from the rat striatum after continuous sucrose density gradient fractionation. The alpha subunit, but not the beta, gamma, or delta subunits, was colocalized with synapsin I, and each subunit showed a distinct distribution pattern in the fractions. To examine further the intracellular distributions of CaM KII isoforms in the same subunit, we established NG108-15 cells stably expressing delta1, delta3, and delta4 isoforms and examined distributions of the delta and gamma isoforms in these cell lines after fractionation. Each of the overexpressed exogenous delta isoforms showed a distinct distribution pattern. The endogenous delta2 was colocalized with the overexpressed delta1, delta3, and delta4 isoforms. However, the endogenous gammaB/gammaC isoforms were not colocalized with the overexpressed delta isoforms. Furthermore, the endogenous delta1 was concentrated in the microsomal fraction from the rat striatum. With the results taken together, it is suggested that CaM KII forms oligomers between isoforms in the same subunit but not in different subunits. The variable domain of CaM KII isoforms might possibly be responsible for targeting to certain intracellular compartments. PMID- 14743432 TI - Regulation of a Ca2+-activated K+ channel by calcium-sensing receptor involves p38 MAP kinase. AB - By using pharmacological and molecular approaches, we previously showed that the G-protein-coupled, extracellular calcium (Ca2+(o))-sensing receptor (CaR) regulates a large-conductance (approximately 140 pS), Ca(2+)-activated K+ channel [IK(Ca); CAKC] in U87 astrocytoma cells. Here we show that elevated Ca2+(o) stimulates extracellular-signal-regulated kinase (ERK1/2) and p38 MAP kinase (MAPK). The effect of high Ca2+(o) on p38 MAPK but not ERK1/2 is CaR mediated, insofar as transduction with a dominant-negative CaR (R185Q) using recombinant adeno-associated virus (rAAV) attenuated the activation of p38 MAPK but not of ERK1/2. p38 MAPK activation by the CaR is likely to be protein kinase C (PKC) independent, in that the pan-PKC inhibitor GF109203X failed to abolish the high Ca2+(o)-induced phosphorylation of p38 MAPK. Consistently with our data on the activation of this kinase, we observed that inhibiting p38 MAPK blocked the activation of the CAKC induced by the specific pharmacological CaR activator NPS R-467. In contrast, inhibiting MEK1 only transiently inhibited the activation of this K+ channel by NPS R-467, despite the continued presence of the antagonist. Similarly to the lack of any effect of the PKC inhibitor on the activation of ERK1/2 and p38 MAPK, inhibiting PKC had no effect on NPS R-467-induced activation of this channel. Therefore, our data show that the CaR, acting via p38 MAPK, regulates a large-conductance CAKC in U87 cells, a process that is PKC independent. Large-conductance CAKCs play an important role in the regulation of cellular volume, so our results have important implications for glioma cell volume regulation. PMID- 14743433 TI - Apoptotic characteristics of cell death and the neuroprotective effect of homocarnosine on pheochromocytoma PC12 cells exposed to ischemia. AB - We recently improved an in vitro ischemic model, using PC12 neuronal cultures exposed to oxygen-glucose deprivation (OGD) for 3 hr in a special device, followed by 18 hr of reoxygenation. The cell death induced in this ischemic model was evaluated by a series of markers: lactate dehydrogenase (LDH) release, caspase-3 activation, presence of cyclin D1, cytochrome c leakage from the mitochondria, BAX cellular redistribution, cleavage of poly (ADP-ribose) polymerase (PARP) to an 85-kDa apoptotic fragment, and DNA fragmentation. The OGD insult, in the absence of reoxygenation, caused a strong activation of the mitogen-activated protein kinase (MAPK) isoforms extracellular regulated kinase (ERK), c-Jun NH2-terminal kinase (JNK), and stress-activated protein kinase (SAPK), also known as p-38. The detection of apoptotic markers and activation of MAPKs during the ischemic insult strongly suggest that apoptosis plays an important role in the PC12 cell death. Homocarnosine, a neuroprotective histidine dipeptide, present in high concentrations in the brain, was found to provide neuroprotection, as expressed by a 40% reduction in LDH release and caspase-3 activity at 1 mM. Homocarnosine reduced OGD activation of ERK 1, ERK 2, JNK 1, and JNK 2 by 40%, 46%, 55%, and 30%, respectively. These results suggest that apoptosis is an important characteristic of OGD-induced neuronal death and that antioxidants, such as homocarnosine, may prevent OGD-induced neuronal death by inhibiting the apoptotic process and/or in relation to the differential attenuation of activity of MAPKs. PMID- 14743434 TI - Increased transcription and activity of glutathione synthase in response to deficiencies in folate, vitamin E, and apolipoprotein E. AB - Oxidative stress is a major contributing factor in neurodegeneration and can arise from dietary, environmental, and genetic sources. Here we examine the separate and combined impact of deprivation of folate and vitamin E, coupled with dietary iron as a prooxidant, on normal mice and transgenic mice lacking apolipoprotein E (ApoE-/- mice). Both mouse strains exhibited increased levels of glutathione when deprived of folate and vitamin E, but a substantial further increase was observed in ApoE-/- mice. To determine the mechanism(s) underlying this increase, we quantified transcription and activity of glutathione synthase (GS). Both normal and ApoE-/- mice demonstrated increased GS activity when deprived of folate and vitamin E. However, transcription was increased only in ApoE-/- mice deprived of folate and vitamin E. These findings demonstrate that deficiency in one gene can result in compensatory up-regulation in a second relevant gene and, furthermore, indicate that compensation for oxidative stress can occur in brain tissue at epigenetic and genetic levels depending on the nature and/or extent of oxidative stress. PMID- 14743435 TI - CD4 T-cell epitopes of human alpha B-crystallin. AB - Of potential importance to multiple sclerosis (MS), oligodendroglial alpha B crystallin is expressed and associated with the myelin sheath at the earliest stage of MS lesion development. We selected T-cell lines specific for human alpha B-crystallin from peripheral blood mononuclear cells (PBMC) of HLA-DR2 homozygous MS patients and found that the alpha B-crystallin-specific T-cells were CD4+ and restricted by DRB1*1501, and expressed Th1 cytokines. The CD4 T-cell epitopes of human alpha B-crystallin were determined by proliferation of alpha B-crystallin specific T-cell lines to 17 20-mer synthetic overlapping peptides spanning the entire molecule of human alpha B-crystallin. It was found that the HLA-DR2 donor derived alpha B-crystallin-specific T-cell lines proliferated to alpha B crystallin peptides 21-40, 41-60, and to a lesser extent, 131-150. These T-cell proliferation responses were associated with intracellular expression of interleukin-2 (IL-2) and secretion of interferon-gamma (IFN-gamma), and tumor necrosis factor-alpha (TNF-alpha). The amino acid sequences of these peptides were compatible with predicted HLA-DR2-restricted binding motifs. PBMC of an early active MS patient proliferated to the epitope-containing peptides significantly better than did those of later stage MS patients or healthy controls. Taken together, these findings suggest that autoreactive alpha B crystallin-specific Th1 cells may have the potential to contribute to MS pathogenesis. PMID- 14743436 TI - Biphasic form of experimental autoimmune neuritis in dark Agouti rats and its oral therapy by antigen-specific tolerization. AB - A new and biphasic form of experimental autoimmune neuritis (EAN) is described in dark agouti rats (DA rats) and is inducible by a single immunization with bovine peripheral nerve myelin (BPM) in complete Freund's adjuvant (DA-EAN). Animals develop a mild episode of disease; after recovery, 66-100% of the rats suffer from a more severe bout of EAN with paraparesis 25-30 days after immunization. By histology, DA-EAN is an inflammatory and demyelinating polyradiculoneuropathy virtually without axonal damage. Demyelination affects mainly spinal roots. This is also reflected by markedly increased F-wave latencies in nerve conduction studies of sciatic nerves. In sciatic nerves, inflammation and demyelination are found only focally and may be the histopathologic basis for conduction failure in some fibers. Immunologic investigations revealed stronger proliferative responses of DA than of Lewis rat lymph node cells to BPM and various peptides derived from the P2 protein. Proliferative and Th1-cytokine responses were particularly pronounced in spleen during the late phase of DA-EAN as compared to the monophasic EAN of Lewis rats. The data suggest that persistent lymphocyte proliferation with secretion of interferon (IFN)-gamma may be relevant for the relapsing course of DA-EAN whereas epitope spreading may explain the increased severity of the second bout of disease. The extended Th1 response in DA rats did not go along with a lack of downregulatory mechanisms, because the second DA-EAN attack was self-limited and splenocytes from DA rats produced considerable amounts of interleukin (IL)-10 and transforming growth factor (TGF)-beta. To substantiate further a functional immunoregulation in DA rats, we modulated DA EAN by antigen-specific oral tolerization, which is known to involve active suppressor mechanisms. Preventive feeding of BPM in combination with cholera toxin (CT) induced a long-lasting resistance to DA-EAN. Even therapeutic administration of BPM or BPM/CT after onset of signs of disease significantly mitigated the further course of disease and prevented development of paraparesis. Because DA-EAN is easily inducible and leads consistently to relapses in most rats, it can be used for studies of immune factors that determine a relapsing course of autoimmunity. Furthermore, DA-EAN may serve as a model for relapsing inflammatory demyelinating polyneuropathies such as chronic inflammatory demyelinating polyneuropathy (CIDP) and for treatment studies. Our findings on effective prevention and therapy of DA-EAN by oral application of myelin/CT corroborate this form of immunomodulation as a treatment strategy for cell mediated processes in chronic inflammatory neuropathies. PMID- 14743437 TI - TNF alpha increases activity of gamma-glutamyl transpeptidase in cultured rat astroglial cells. AB - To investigate the presence of gamma-glutamyl transpeptidase (gammaGT) in brain cells, cultures enriched for astroglial cells, neurons, oligodendroglial cells, and microglial cells were studied. Astroglial cultures contained a specific gammaGT activity of 2.3 +/- 0.9 nmol/min/mg protein. A similar specific gammaGT activity was measured for oligodendroglial cultures, whereas microglial cells and neurons contained less than 30% of the specific gammaGT activity of astroglial cultures. The activity of gammaGT in astroglial cultures was elevated strongly by the presence of tumor necrosis factor-alpha (TNFalpha) in a time- and concentration-dependent manner. Maximal activity of gammaGT was observed after incubation of astroglial cultures for 3 days with 30 ng/mL TNFalpha. Under these conditions the specific gammaGT activity was increased by threefold compared to controls. Presence of the gammaGT-inhibitor acivicin completely inhibited gammaGT activity both in TNFalpha-treated and in control cells. In addition, the increase in astroglial gammaGT activity after application of TNFalpha was prevented completely by the presence of the protein synthesis inhibitor cycloheximide. gammaGT is involved in extracellular processing of glutathione (GSH) that is exported by astroglial cells. After TNFalpha-treatment the concentration of GSH in the medium of astroglial cells was reduced significantly compared to control cells. In conclusion, the data presented demonstrate that TNFalpha stimulates gammaGT synthesis in astroglial cells and thereby improves the capacity to process GSH exported by these cells. PMID- 14743438 TI - A non-peptide substance P antagonist (CP-96,345) inhibits morphine-induced NF kappa B promoter activation in human NT2-N neurons. AB - Opioids and the neuropeptide substance P (SP) modulate the expression of inflammatory cytokines and chemokines, which are under the control of nuclear factor kappaB (NF-kappaB). We investigated whether the neurokinin-1 receptor (SP receptor) pathway is biologically involved in morphine-mediated modulation of NF kappaB promoter activation in a human neuronal cell line (NT2-N) that expresses both the mu-opioid receptor (MOR) and the SP receptor. Morphine significantly enhanced NF-kappaB promoter-directed luciferase activity in NT2-N neurons. DAMGO, a selective mu-opioid receptor agonist, also induced NF-kappaB promoter activation. The induced activation of NF-kappaB promoter by morphine or DAMGO was abolished not only by naltrexone (a opioid receptor antagonist) and CTAP (a selective, competitive mu-opioid receptor antagonist), but also by CP-96,345, a non-peptide SP receptor antagonist. Investigation of the mechanism responsible for morphine-induced activation of NF-kappaB promoter in NT2-N neurons demonstrated that morphine activates the SP promoter and induces SP expression in these cells. We also observed that SP activated NF-kappaB promoter and that CP 96,345 downregulated the expression of endogenous SP. Furthermore, dual immunofluorescent labeling revealed that there is co-expression of NK-1R and MOR in the processes of NT-2N neurons. These results suggest that morphine, by activating MOR, engages a positive feedback loop between NK-1R and SP. Activation of NK-1R could then impact NF-kappaB expression and therefore may be an important participant in the effect of morphine on immune responses in the central nervous system. PMID- 14743439 TI - Enhanced inflammatory response in neural tubes of embryos derived from diabetic mice exposed to a teratogen. AB - Exposure of embryos to the teratogen cyclophosphamide (CP) and maternal diabetes is linked to pathogenesis of neural tube defects during development. Maternal diabetes aggravates the teratogen-induced inflammatory reaction leading to increased risk of neural tube defects in mouse embryos. The inflammatory reaction in the developing neural tube has been characterized by the presence of activated amoeboid microglia/brain macrophages and altered expression levels of cytokines. Although there were no obvious anomalies observed in the neural tubes of embryos from CP-treated non-diabetic mice, the frequency of neural tube defects was increased significantly in embryos of CP-treated diabetic mice. Moreover, there were more activated amoeboid microglia in the forebrain of CP-treated diabetic embryos compared to that in CP-treated non-diabetic mice. The expression of cytokines such as tumor necrosis factor-alpha (TNF-alpha) and transforming growth factor-beta1 (TGF-beta1) in the fetal brain of normal and diabetic embryos was induced in the neural tubes after CP treatment. Furthermore, the mRNA expression levels of both genes were increased markedly in the neural tube of CP-treated diabetic embryos compared to that of CP-treated non-diabetic embryos as measured by quantitative real-time PCR. Immunohistochemically, more TNF-alpha- and TGF beta1-positive cells, which included neurons and amoeboid microglia, were detected in CP-treated diabetic embryos than in CP-treated normal embryos. Maternal diabetes aggravates teratogen-induced inflammation, which is characterized in the developing neural tube by increased amoeboid microglia and enhanced expression of inflammatory cytokines. Although a definite link has yet to be elucidated, it is suggested that the increased rate of neural tube defects observed in CP-treated diabetic embryos may be due to upregulation of proinflammatory cytokines caused by maternal diabetes. PMID- 14743440 TI - Chronic exposure to aluminum in drinking water increases inflammatory parameters selectively in the brain. AB - A link between aluminum (Al) exposure and age-related neurological disorders has long been proposed. Although the exact mechanism by which the metal may influence disease processes is unknown, there is evidence that exposure to Al causes an increase in both oxidative stress and inflammatory events. These processes have also been suggested to play a role in Alzheimer's disease (AD), and exposure to the metal may contribute to the disorder by potentiating these events. Al lactate (0.01, 0.1, and 1 mM) in drinking water for 10 weeks increased inflammatory processes in the brains of mice. The lowest of these levels is in the range found to increase the prevalence of AD in regions where the concentrations of the metal are elevated in residential drinking water (Flaten [2001] Brain Res. Bull. 55:187 196). Nuclear factor-kappaB as well as tumor necrosis factor-alpha (TNF-alpha) and interleukin 1alpha (IL-1alpha) levels were increased in the brains of treated animals. The mRNA for TNF-alpha was also up-regulated following treatment. Enhancement of glial fibrillary acidic protein levels and reactive microglia was seen in the striatum of Al-treated animals. The level of amyloid beta (Abeta40) was not significantly altered in the brains of exposed animals. Insofar as no parallel changes were observed in the serum or liver of treated animals, the proinflammatory effects of the metal may be selective to the brain. Al exposure may not be sufficient to cause abnormal production of the principal component of senile plaques directly but does exacerbate underlying events associated with brain aging and thus could contribute to progression of neurodegeneration. PMID- 14743441 TI - Retinoic acid-mediated induction of neurons and glial cells from human umbilical cord-derived hematopoietic stem cells. AB - Recent studies reporting trans-differentiation of mononucleated cells derived from human umbilical cord blood into neuronal cells aroused interest among investigators for their clinical implication and significance in regenerative medicine. In the present study, purified populations of hematopoietic stem cells were isolated via magnetic bead sorting and fluorescence-activated cell sorter (FACS) using a specific CD133 antibody, a cell type-specific marker for hematopoietic stem cells, and grown in culture in the presence of retinoic acid (RA). CD133+ hematopoietic stem cells expressed neuronal and glial phenotypes after RA treatment. RT-PCR analysis indicated that the RA treated CD133+ cells expressed mRNA transcripts for ATP-binding cassettes transporter ABCG2 (a universal stem cell marker), nestin (a specific cell type marker for neural stem cells), Musashi1 (a specific marker for neural stem cells) and RA receptors (RAR) including RAR-alpha, RAR-beta, and retinoid X receptor (RXR)-gamma. RA-treated CD133+ cells expressed mRNA transcripts for neuron-specific markers neurofilament proteins (NF-L, -M, -H) and synaptophysin as determined by RT-PCR, structural proteins characteristic of neurons including tubulin beta III and neuron specific enolase (NSE) by Western blot, and neuron-specific markers NeuN and microtubule associated protein-2 (MAP2) by immunocytochemistry. RA-treated CD133+ cells also expressed the astrocyte-specific marker glial fibrillary acidic protein (GFAP), as demonstrated by RT-PCR, Western blot, and immunocytochemistry. In addition, RA treated CD133+ cells expressed cell type-specific markers for oligodendrocytes including myelin basic protein (MBP) as shown by RT-PCR, proteolipid protein (PLP) by Western blot analysis, and cyclic nucleotide phosphodiesterase (CNPase) by immunostaining. Upregulated expression of several basic helix-loop-helix (bHLH) transcription factors important for early neurogenesis, including Otx2, Pax6, Wnt1, Olig2, Hash1 and NeuroD1, was also demonstrated in CD133+ cells after RA treatment. These results indicate that human cord blood-derived CD133+ hematopoietic stem cells could trans-differentiate into neural cell types of neuron-like cells, astrocytes, and oligodendrocytes by RA treatment. PMID- 14743442 TI - Noise trauma alters D-[3H]aspartate release and AMPA binding in chinchilla cochlear nucleus. AB - Exposure of adults to loud noise can overstimulate the auditory system, damage the cochlea, and destroy cochlear nerve axons and their synaptic endings in the brain. Cochlear nerve loss probably results from the death of cochlear inner hair cells (IHC). Additional degeneration in the cochlear nucleus (CN) is hypothesized to stem from overstimulation of the system, which may produce excitotoxicity. This study tested these predictions by exposing one ear of anesthetized adult chinchillas to a loud noise, which damaged the ipsilateral cochlea and induced degeneration in the glutamatergic cochlear nerve. During the first postexposure week, before cochlear nerve axons degenerated, glutamatergic synaptic release in the ipsilateral CN was elevated and uptake was depressed, consistent with hyperactivity of glutamatergic transmission and perhaps with the operation of an excitotoxic mechanism. By 14 days, when cochlear nerve fibers degenerated, glutamatergic synaptic release and uptake in the CN became deficient. By 90 days, a resurgence of transmitter release and an elevation of AMPA receptor binding suggested transmission upregulation through plasticity that resembled changes after mechanical cochlear damage. These changes may contribute to tinnitus and other pathologic symptoms that precede and accompany hearing loss. In contrast, the other ear, protected with a silicone plug during the noise exposure, exhibited virtually no damage in the cochlea or the cochlear nerve. Altered glutamatergic release and AMPA receptor binding activity in the CN suggested upregulatory plasticity driven by signals emanating from the CN on the noise exposed side. PMID- 14743443 TI - Anti-Abeta: The good, the bad, and the unforeseen. AB - Alzheimer's disease (AD) is characterized in part by the deposition of amyloid beta protein (Abeta) in compact fibrillar plaques. These structures can induce an innate immune response in the brain, which triggers progressive inflammation, neuronal loss, and further acceleration of Abeta plaque formation. Compared with the case in normal individuals, the T and B lymphocytes in AD patients and murine models are hyporesponsive to Abeta. However, depending on the route of delivery, tolerance can be overcome by vaccination, with the induction of an anti-Abeta mediated immune response. Through mechanisms that are incompletely understood, immunized APP transgenic animals show markedly reduced Abeta deposition, preservation of normal neuronal architecture, and improved performance in memory and spatial learning tasks. In human trials, Abeta vaccination stabilized cognition and slowed the progression of dementia. Neuropathologic examination of a vaccinated subject showed reduced cortical Abeta without changes in other AD associated pathology. However, in some patients, vaccination induced severe meningoencephalitis, causing the trial to be terminated. Thus, vaccination appears to activate both beneficial and deleterious anti-Abeta immunity, suggesting that the vaccine can have potent clinical utility if an appropriate immunologic response can be generated. PMID- 14743445 TI - Neuritogenesis induced by vasoactive intestinal peptide, pituitary adenylate cyclase-activating polypeptide, and peptide histidine methionine in SH-SY5y cells is associated with regulated expression of cytoskeleton mRNAs and proteins. AB - Vasoactive intestinal peptide (VIP) and the related peptides pituitary adenylate cyclase-activating polypeptide (PACAP) and peptide histidine methionine (PHM) are known to regulate proliferation and/or differentiation in normal and tumoral cells. In this study, neuritogenesis in human neuroblastoma SH-SY5Y cells cultured in serum-free medium was induced by VIP, PACAP, and PHM. The establishment of this process was followed by the quantification of neurite length and branching and the expression of neurofilament mRNAs, neurofilament proteins, and other cytoskeletal protein markers of neuronal differentiation: neuron-specific MAPs and beta-tubulin III. Neurite length and branching and the expression of most markers tested were increased by VIP and PACAP in a similar, although slightly different, fashion. In contrast, neuritic elongation induced by PHM was correlated with neither an increase in branching or neurofilament mRNAs nor a clear change in the expression of cytoskeleton proteins, with the exception of the stimulation by PHM of doublecortin, a microtubule-associated marker of migrating neuroblasts. These findings are the first evidence from a human neuron like cell line for 1) a direct regulation of the metabolism of neurofilaments by VIP and PACAP and 2) the induction by PHM of neuritic processes of an apparent immature character. PMID- 14743444 TI - Identification of a Hoxd10-regulated transcriptional network and combinatorial interactions with Hoxa10 during spinal cord development. AB - Hoxd10 is expressed in the posterior spinal cord and hindlimbs of the mouse. Hoxd10, along with other Hox transcription factors, is thought to regulate the activity of genes involved in nervous system patterning and motor neuron development, but little is known about the downstream targets regulated by this gene. cDNA microarrays were used to investigate the transcriptional network regulated by Hoxd10 in homozygous knockout animals. Sixty-nine genes were identified with altered expression levels in mutant spinal cords. Among these were genes involved in such diverse cellular events as cellular communication, cell cycle control, development and differentiation, and neuronal survival. The expression of some of these genes was investigated using reverse transcriptase polymerase chain reaction (RT-PCR) and in situ hybridization. Nine genes showed changes in expression of the same sign and similar magnitude using RT-PCR in Hoxd10 single mutant animals, with additional changes in expression seen in Hoxa10/Hoxd10 double mutant animals. In situ hybridization studies also demonstrated changes in expression consistent with microarray results. Analysis of putative promoter regions for Hox protein binding sites suggested that some genes may be direct Hoxd10 targets, whereas others likely are regulated through intermediate steps. Using cDNA microarrays to study a single gene knockout during critical developmental stages has identified a large number of genes regulated by Hoxd10, many of which would not have been approached as candidates for Hox gene regulation based on function or expression. PMID- 14743446 TI - Patterns of Jagged1, Jagged2, Delta-like 1 and Delta-like 3 expression during late embryonic and postnatal brain development suggest multiple functional roles in progenitors and differentiated cells. AB - The Notch-DSL signaling system, consisting of multiple receptors and ligands, inhibits neurogenesis and promotes gliogenesis during embryonic development, but the specific function of the various ligands and receptors at later developmental stages are unknown. Here, we examined the expression pattern of four Delta, Serrate and Lag-2 (DSL) ligands, Jagged1, Jagged2, Delta-like1 (Dl1) and Delta like 3 (Dl3), in late embryonic and postnatal rat brain by in situ hybridization. In late embryos, Jagged1, Dl1 and Dl3 mRNAs were present in the periventricular germinal epithelia, but this expression diminished during postnatal ages. Jagged1 mRNA was also expressed in the inner aspect of the dentate gyrus at early postnatal times. Dl3 was detectable in the external granule cell layer (EGL) of the cerebellum, another site of postnatal neurogenesis. Jagged2 mRNA was expressed in virtually all postnatal neurons. Jagged1 mRNA was highly expressed in several brain nuclei during postnatal development, with lower levels of expression in other grey matter regions. In white matter, Dl1 and Dl3 mRNAs were expressed during the first week of postnatal development but only the expression of Dl1 mRNA persisted through the second week. Dl1 mRNA was present at lower levels throughout grey matter during the first few weeks of development. Jagged1 mRNA was expressed in blood vessels, choroid plexus, and menninges throughout development and in the adult. Jagged2 mRNA was transiently expressed in cerebral blood vessels and choroid plexus during the first postnatal week. Taken together, these results support multiple and differing roles for the various ligands during and after central nervous system (CNS) development. PMID- 14743447 TI - TIEG1 facilitates transforming growth factor-beta-mediated apoptosis in the oligodendroglial cell line OLI-neu. AB - Transforming growth factor-beta (TGF-beta) plays an important role during the period of developmental cell death in the nervous system. Using the oligodendroglial precursor cell line OLI-neu, we have previously established an in vitro system to analyze TGF-beta-mediated cell death on the molecular level. We could show that the Kruppel-like Zn-finger transcription factor TIEG1 was up regulated after TGF-beta stimulation of OLI-neu cells and mimicked TGF-beta effects in these cells; i.e., overexpression of TIEG1 in OLI-neu cells induced apoptosis as shown by apoptosis ELISA, DNA fragmentation, and caspases-3 activation. The apoptotic pathway seemed to be initiated by repressing the expression of the antiapoptotic protein Bcl-XL. In contrast, the reporter activity of a SMAD consensus promoter was induced, whereas the promoter activity of the inhibitory SMAD7 was reduced, suggesting that SMAD-dependent TGF-beta responses, such as TGF-beta-induced apoptosis, are enhanced in the presence of TIEG1. PMID- 14743448 TI - ADAM (a disintegrin and metalloprotease) 12 is expressed in rat and human brain and localized to oligodendrocytes. AB - ADAM12 is a member of the large family of multidomain metalloprotease disintegrins which possess cell-binding and metalloprotease properties. Typically, ADAM12 is expressed in mesenchymal cells, developing and regenerating heart and skeletal muscle, bone as well as in certain tumours. This report shows by means of reverse transcriptase-polymerase chain reaction (RT-PCR) and immunohistochemistry that the protease ADAM12 is detectable in human and rat brain tissue as well as in cultured cells derived from rat brain. With the exception of a very few immunopositive pyramidal neurons in the developing rat brain, the cellular localization of ADAM12 was exclusively confined to oligodendroglial cells. Thus, ADAM12 may be regarded a new suitable marker for this cell type. PMID- 14743450 TI - Altered localization of Cav1.2 (L-type) calcium channels in nerve fibers, Schwann cells, odontoblasts, and fibroblasts of tooth pulp after tooth injury. AB - We have determined the localization of Cav1.2 (L-Type) Ca2+ channels in the cells and nerve fibers in molars of normal or injured rats. We observed high levels of immunostaining of L-type Ca2+ channels in odontoblast cell bodies and their processes, in fibroblast cell bodies and in Schwann cells. Many Cav1.2-containing unmyelinated and myelinated axons were also present in root nerves and proximal branches in coronal pulp, but were usually missing from nerve fibers in dentin. Labeling in the larger fibers was present along the axonal membrane, localized in axonal vesicles, and in nodal regions. After focal tooth injury, there is a marked loss of Cav1.2 channels in injured teeth. Immunostaining of Cav1.2 channels was lost selectively in nerve fibers and local cells of the tooth pulp within 10 min of the lesion, without loss of other Cav channel or pulpal labels. By 60 min, Cav1.2 channels in odontoblasts were detected again but at levels below controls, whereas fibroblasts were labeled well above control levels, similar to upregulation of Cav1.2 channels in astrocytes after injury. By 3 days after the injury, Cav1.2 channels were again detected in nerve fibers and immunostaining of fibroblasts and odontoblasts had returned to control levels. These findings provide new insight into the localization of Cav1.2 channels in dental pulp and sensory fibers, and demonstrate unexpected plasticity of channel distribution in response to nerve injury. PMID- 14743449 TI - Protein kinase A and calcium/calmodulin-dependent protein kinase II regulate glycine and GABA release in auditory brain stem nuclei. AB - We reported previously that unilateral cochlear ablation (UCA) in young adult guinea pigs induced protein kinase C (PKC)-dependent plastic changes in the electrically evoked release of exogenous [14C]glycine ([14C]Gly) or [14C]-gamma aminobutyric acid ([14C]GABA) in several brain stem auditory nuclei. The present study assessed whether such changes depended on protein kinase A (PKA) and calcium/calmodulin-dependent protein kinase II (CaMKII). In the major subdivisions of the cochlear nucleus (CN) and the main nuclei of the superior olivary complex (SOC) dissected from intact animals, dibutyryl-cyclic adenosine monophosphate (DBcAMP) (0.2 mM), a PKA activator, elevated release by 1.6-2.3 fold. The PKA inhibitor, H-89 (2 microM), did not alter the release but blocked the stimulatory effects of DBcAMP. These findings suggested that PKA could positively regulate glycinergic and GABAergic release. After UCA, PKA regulation declined and failed in the ventral CN but persisted in the SOC nuclei. After 145 postablation days, H-89 reversed elevations of [14C]GABA release in the medial nucleus of the trapezoid body (MNTB). A CaMKII inhibitor, KN-93, reversed depressions of [14C]Gly release in the DCN. Thus, the postablation plasticities in these nuclei probably depended on PKA or CaMKII. Both H-89 and KN-93 depressed [14C]Gly release in the lateral superior olive (LSO) and ipsilateral medial superior olive (MSO), suggesting that either kinase was used by endogenous mechanisms in these nuclei to upregulate glycinergic release. In contrast, KN-93 elevated [14C]GABA release in the contralateral MNTB, suggesting a downregulatory action of CaMKII, an action opposite to that of PKA. PMID- 14743451 TI - Suppression of proliferation and neurite extension of human neuroblastoma SH-SY5Y cells on immobilized Psathyrella velutina lectin. AB - Glycoproteins from mammalian brain tissues contain unique N-linked oligosaccharides terminating with beta-N-acetylglucosamine residues. Lectin blot analysis of membrane glycoprotein samples from human neuroblastoma SH-SY5Y cells showed that several protein bands bind to Psathylera velutina lectin (PVL), which interacts with beta-N-acetylglucosamine-terminating oligosaccharides. No lectin positive bands were detected by digestion with jack bean beta-N-acetyl hexosaminidase or N-glycanase before incubation with the lectin, indicating that the cells contain beta-N-acetylglucosamine-terminating N-linked oligosaccharides. When cells were cultured in dishes with different concentrations of PVL, the cell proliferation was inhibited in a dose-dependent manner. Similarly, the neurite extension, which was stimulated with nerve growth factor, was also inhibited in a manner dependent on the lectin dose. Cell proliferation and neurite extension were recovered by the addition of 10 mM N-acetylglucosamine into the medium. Immunoblot analysis of the activation of mitogen-activated protein (MAP) kinases and protein kinase C revealed that phosphorylation of 42-kDa and 44-kDa MAP kinases and 80-kDa protein kinase C are inhibited when SH-SY5Y cells are cultured in PVL-coated dishes, but are restored by the addition of the haptenic sugar into the medium, indicating that MAP kinase and protein kinase C pathways are inhibited by interaction with immobilized PVL. These results indicate that beta-N acetylglucosamine-terminating N-linked oligosaccharides expressed on neural cells can induce intracellular signals upon binding to extracellular receptors, and are important for growth regulation of neural cells. PMID- 14743452 TI - Treatment with the neurosteroid dehydroepiandrosterone promotes recovery of motor behavior after moderate contusive spinal cord injury in the mouse. AB - The neurosteroid dehydroepiandrosterone (DHEA) has neuroprotective properties after ischemic and excitatory insults to the brain. In the developing embryo, it is produced in discrete regions of the central nervous system (CNS), where it specifically promotes axonal growth of differentiated neurons. To test if DHEA could be beneficial after spinal cord injury (SCI), we used a model of moderate contusive SCI developed and characterized in the mouse. Immediately after surgery, we applied treatment with DHEA or with vehicle only and compared treatment groups (n = 12 in each group) over a 42-day period. Locomotor recovery was assessed in an open field using a standardized 21-point scale, according to gait analysis on paw print recordings and using foot fault analyses on an inclined ladder beam. The DHEA-treated group showed improved function compared to vehicle-treated animals in these tests. More strikingly, DHEA enhanced recovery of left-right coordination and fine motor control. In an attempt to correlate functional recovery with spinal cord neuropathology in the different experimental groups, we studied the area of spared white matter at the epicenter and reactive gliosis/scar formation 42 days post-injury (DPI). DHEA significantly increased the area of white matter spared at the epicenter and reduced the area of reactive gliosis surrounding the lesion. These data demonstrate the effectiveness of DHEA in promoting functional recovery in the adult murine injured spinal cord. PMID- 14743453 TI - CRE-mediated gene transcription in the peri-infarct area after focal cerebral ischemia in mice. AB - Cyclic AMP response element binding protein (CREB) is a transcription factor expressed constitutively primarily in neurons and is activated by phosphorylation at Ser(133) residue. CREB mediates expression of several neuroprotective proteins, including B-cell CLL/lymphoma 2 (BCL-2) and brain-derived neurotrophic factor (BDNF). Although phosphorylation of CREB after ischemia has been investigated extensively, CRE-mediated gene transcription after ischemia is not as well studied. We investigated temporal changes in CRE-mediated gene transcription in the cerebral cortex after focal ischemia in transgenic mice with a CRE-lacZ reporter gene. In the ischemic core, X-gal-positive cells, which reflected expression of the CRE-lacZ reporter gene, were observed rarely at any time point, though transient phosphorylation of CREB was detected. In contrast, the peri-infarct area showed a persistent increase in the number of X-gal positive cells, of which more than half were positive for neuronal nuclei (NeuN). Our results suggest that CRE-mediated gene transcription, the pattern of which is not always consistent with that of CREB phosphorylation, occurs primarily in neurons in the peri-infarct area after focal cerebral ischemia and may be a neuroprotective response against ischemic insult. PMID- 14743454 TI - Sulfate- and size-dependent polysaccharide modulation of AMPA receptor properties. AB - Previous work found evidence that alpha-amino-3-hydroxy-5-methylisoxazole-4 propionic acid (AMPA)-type glutamate receptors interact with and are functionally regulated by the glycosaminoglycan heparin. The present study tested whether dextran species affect ligand binding, channel kinetics, and calcium permeability of AMPA receptors. Dextran sulfate of 500 kDa markedly reduced high affinity [3H]AMPA binding in solubilized hippocampal membranes. In isolated receptors reconstituted in a lipid bilayer, the same dextran sulfate prolonged the lifetime of open states exhibited by AMPA-induced channel fluctuations. The large polysaccharide further changed the single channel kinetics by increasing the open channel probability five- to sixfold. Such modulation of channel activity corresponded with enhanced levels of calcium influx as shown in hippocampal neurons loaded with Fluo3AM dye. With an exposure time of <1 min, AMPA produced a dose-dependent increase in intracellular calcium that was blocked by 6-cyano-7 nitroquinoxaline-2,3-dione disodium (CNQX). Dextran sulfate, at the same concentration range that modified ligand binding (EC50 of 5-10 nM), enhanced the AMPA-induced calcium influx by as much as 60%. The enhanced influx was blocked by CNQX, although unchanged by the N-methyl-D-aspartate (NMDA) receptor antagonist AP5. Confocal microscopy showed that the increase in calcium occurred in neuronal cell bodies and their processes. Interestingly, smaller 5-8-kDa dextran sulfate and a non-sulfated dextran of 500 kDa had little or no effect on the binding, channel, and calcium permeability properties. Together, these findings suggest that synaptic polysaccharide species modulate hippocampal AMPA receptors in a sulfate- and size-dependent manner. PMID- 14743455 TI - Amino- and carboxyl-terminal mutants of presenilin 1 cause neuronal cell death through distinct toxic mechanisms: Study of 27 different presenilin 1 mutants. AB - Presenilin (PS)1 and its mutants, which consist of the N-terminal and C-terminal fragments, cause certain familial forms of Alzheimer's disease (FAD). Our earlier studies found that FAD-linked M146L-PS1 causes neuronal cell death through nitrogen oxide synthase (NOS) and that FAD-linked N141I-PS2, another member of the PS family, causes neuronal cell death through NADPH oxidase. In this study, we examined 27 different FAD-linked mutants of PS1, and found that PS1 mutants with mutations in the N-terminal fragment caused NOS inhibitor (NOSI)-sensitive neuronal cell death; in contrast, the PS1 mutants with mutations in the C terminal fragment caused NOSI-resistant neuronal cell death. The former toxicity was resistant to the specific NADPH oxidase inhibitor apocynin and was inhibited by Humanin (HN), a newly identified neuroprotective factor against Alzheimer's disease (AD)-relevant insults, but not by insulin-like growth factor-I (IGF-I). In contrast, the latter toxicity was sensitive to apocynin and inhibited by both IGF-I and HN. This study indicates for the first time that N- and C-terminal fragment PS1 mutants can generate distinct neurotoxic signals, which will provide an important clue to the understanding of the entire array of neurotoxic signals generated by FAD-causative mutations of PS1. PMID- 14743456 TI - Semi-chronic increase in striatal level of 3,4-dihydroxyphenylacetaldehyde does not result in alteration of nigrostriatal dopaminergic neurones. AB - This work was carried out to evaluate the potential in vivo toxicity of 3,4 dihydroxyphenylacetaldehyde (DOPAL), an aldehyde formed from dopamine by monoamine oxidase (MAO) that is oxidised mainly to 3,4-dihydroxyphenylacetic acid (DOPAC) by brain aldehyde dehydrogenases (ALDH). In this study, male Sprague Dawley rats were treated with levodopa (L-dopa)-benserazide, which increases DOPAL production by MAO, and disulfiram, an irreversible inhibitor of ALDH, which reduces the formation of DOPAC from DOPAL. An acute systemic intraperitoneal (i.p.) injection of 100 mg/kg disulfiram and L-dopa-benserazide (100 mg/kg + 25 mg/kg, 24 hr later) significantly increased DOPAL striatal level. A 30-day treatment with disulfiram (100 mg/kg i.p., once every 2 days) and L-dopa benserazide (100 mg/kg + 25 mg/kg, two times/day) did not affect either indexes used to assess integrity of the nigrostriatal dopaminergic neurones (i.e., the striatal content in dopamine and binding to the vesicular monoamine transporter on striatal membranes). These results do not evidence any deleterious effect of DOPAL and argue against toxicity of L-dopa therapy. PMID- 14743457 TI - Kynurenic acid synthesis in cerebral cortical slices of rats with progressing symptoms of thioacetamide-induced hepatic encephalopathy. AB - Increased ammonia is a major pathogenic factor in hepatic encephalopathy (HE), a neurologic syndrome associated with glutamatergic dysfunction. Previous studies have shown that in rat cerebral cortical slices or a glia-derived cell line, acute treatment with ammonia in vitro and in vivo inhibits the production of a broad-spectrum antagonist of excitatory amino acid receptors, kynurenic acid (KYNA). The present study analyzed KYNA synthesis in cerebral cortical slices obtained from rats with progressing HE symptoms accompanying acute liver failure induced by one, two, or three intraperitoneal administrations of thioacetamide (TAA) at 24-hr intervals. KYNA synthesis was found decreased to 83% of control 24 hr after one administration of TAA and unaffected after two TAA injections, when moderate hyperammonemia was associated by metabolic and bioelectric activation of the central nervous system, but was not accompanied by typical HE symptoms. KYNA synthesis was elevated to 155% of control after three TAA administrations, a period in which the rats showed advanced HE symptoms including stupor or coma. KYNA synthesis at the advanced HE stage was inhibited by glutamate in a degree comparable to that observed in control slices. The elevation of KYNA synthesis was associated with increased activity of a kynurenine aminotransferase (KAT) isomer, KAT-II. KYNA synthesis did not differ from control 21 days after the third TAA administration when HE symptoms receded. The results suggest that alterations of KYNA synthesis may contribute to the imbalance between neural excitation and inhibition at different stages of HE. PMID- 14743458 TI - Graded reoxygenation with chemical inhibition of oxidative phosphorylation improves posthypoxic recovery in murine hippocampal slices. AB - Rapid and complete tissue reoxygenation is a prime goal of present stroke therapy. However, reoxygenation may trigger detrimental cascades that partially antagonize beneficial effects. It was our goal to investigate selective grading of reoxygenation with targeting of single mitochondrial complexes in murine hippocampal slices. Population spike amplitude (PSAP) and NADH were measured during hypoxic hypoxia (15 min) and recovery (45 min). With onset of reoxygenation, slices were treated for different times with amobarbital (1 mM), malonate (2 mM), or cyanide (1 mM), inhibitors of mitochondrial complex I, II, or IV, respectively. Other slices were treated with nicotinamide (1 mM). Posthypoxic recovery of PSAP increased from 32% +/- 43% of onset in control slices to 52% +/- 59% (P <.05) upon treatment with amobarbital for 1 min and to 62% +/- 37% (P <.05) upon treatment with malonate. With nicotinamide, posthypoxic recovery improved to 73% +/- 25% (P <.05). Oxidation of NADH was prolonged upon treatment with amobarbital, whereas no change in NADH oxidation was observed with malonate and nicotinamide. Thus, grading of reoxygenation with selective targeting of mitochondrial complex I or II but not of complex IV improves outcome upon reoxygenation in murine hippocampal slices. PMID- 14743459 TI - Development of prostate cancer treatment: the good news. AB - Prostate cancer is the most commonly diagnosed cancer in American men representing one-third of all new cancer cases each year. This translates into one out of every six American men being diagnosed with prostate cancer over the course of their lifetimes. Over 31,000 of these men die each year from prostate cancer. Before the 1980's, 50% of men were diagnosed with widespread metastatic disease and there were few therapeutic choices for patients. The good news for patients is that, over the last 30 years there have been significant advances in detection and prognostication as well as major improvements in the surgical, radiation, and medical oncological management of prostate cancer. This review describes the evolution of these therapeutic modalities for prostate cancer. This evolution has been driven by the explosion of knowledge concerning cancer in general and in the specific biology of prostate cancer in particular over the last 30 years. This knowledge has been obtained by concentrating human and financial resources in organ specific studies of the prostate. The end result of this effort is that, today, 85% of new prostate cancer cases are diagnosed at local and regional stages and the 5-year relative prostate cancer survival rate has increased by 20% since 1985. In addition, the therapeutic approach to prostate cancer can now be individualized based on the characteristics of the patient's disease. Finally, recent data suggest that the death rate from prostate cancer is decreasing by approximately 4% per year since 1994. Further good news for patients is that new discoveries about the biology of prostate cancer are rapidly being translated into new therapies, a large number of which are currently being tested in clinical trials. Continued allocation of appropriate human and material resources should yield new, more effective therapies for prostate cancer that will further impact patient quality of life and survival in the 21st century. PMID- 14743460 TI - Development of new prostate specific monoclonal antibodies. AB - BACKGROUND: Despite the need for new prostate-specific diagnostic and therapeutic targets, very few unique prostate (cancer) specific antigens have been characterized. Monoclonal antibody (mAb) technology is a powerful tool to identify specific antigenic markers, which could be potential targets for cancer diagnostics or therapy. METHODS: Splenocytes from mice immunized with prostate cancer (PCa) homogenates of different origin were fused using standard techniques. Employing a differential high-throughput screening method followed by immediate screening in immunohistochemistry (IHC) a large number of hybridomas were screened for prostate (cancer) specificity. RESULTS: From 25 successful fusions approximately 300 clones were identified excreting PCa-reactive antibodies. Subsequent immunohistochemical fine-specificity analysis reduced this number to 26. Eventually, after extensive fine-specificity analysis, the number of mAbs appearing to define prostate-specific antigenic structures that might serve as new diagnostic or therapeutic targets was reduced to three. CONCLUSIONS: Using mAb technology combined with a high throughput screening method we have developed three mAbs (1.8, 2.26, and 3.10) directed against prostate associated antigens that might identify potential new therapeutic targets. PMID- 14743461 TI - Characteristics of prostate cancers detected at low PSA levels. AB - BACKGROUND: When age-referenced PSA levels as recommended by Oesterling et al.1 were used as a biopsy criterion, only 25% of the cancers detected in a population based PSA Screening Project were organ-confined. This observation led to the decision to use low PSA levels as the sole indication for biopsy. Since 1995 age referenced PSA levels of 1.25-3.25 ng/ml have been used in combination with a percentage free PSA cutoff of 18%. This PSA cutoff reduction led to a statistically significant migration to lower pathological stages with a decreased prostate cancer mortality in the years 1996-2001. However, concerns have been raised that screening with low PSA levels may detect clinically insignificant cancers. MATERIALS AND METHODS: We evaluated prostate cancer patients with low PSA levels in terms of heterogeneity, clinical significance, multifocality, and tumor biology including ploidy and proliferation index. RESULTS: Concerning heterogeneity the Gleason score of the needle biopsy failed to predict the Gleason score of the radical prostatectomy specimen in nearly 40% of prostate cancer patients; regarding multifocality 65% of patients with low PSA levels showed multifocal lesions and 36% exhibited tetraploid DNA distribution; more than 50% of tetraploid tumors were found in patients with tumor volumes of less than 0.5 cm(3). Ploidy correlated with the Ki-67 proliferation index, but not with tumor volume. CONCLUSIONS: These results demonstrate that small prostate cancers with low PSA levels and low tumor volumes exhibit all features of prostate cancers with higher tumor volumes and show the characteristics of malignant cancers, i.e., multifocality, tetraploidy, and high proliferative activity. PMID- 14743462 TI - Membrane properties of single muscle cells of the rhabdosphincter of the male urethra. AB - BACKGROUND: The electrophysiological properties of myoblast cultures established from the human and porcine rhabdosphincter (RS) and porcine lower limb muscle (LLSKM) were studied to elucidate their potential for tissue engineering applications in the lower urinary tract. METHODS: Muscle biopsies were collected from the prostatic part of the RS, the RS of male pigs, and the porcine LLSKM. Ion channels were studied by means of the patch-clamp technique. RESULTS: Only one subtype each of voltage gated Na+ and Ca2+ channels was observed in porcine RS and LLSKM. Two types of voltage gated Ca2+ channels were identified in human RS cells. The porcine RS and LLSKM myoblasts displayed similar fusion competence. CONCLUSIONS: Porcine RS and LLSKM myoblasts and human RS and human skeletal muscle cells show a high degree of similarity. Injection of autologous skeletal muscle myoboblasts in the lower urinary tract might, therefore, represent a promising approach to treat stress incontinence after radical prostatectomy. PMID- 14743463 TI - Long-term morbidity and mortality of transurethral prostatectomy: a 10-year follow-up. AB - BACKGROUND: Long-term complication and reoperation rates in 1,211 patients undergoing TURP for symptomatic BPH between 1988 and 1991 were evaluated after a follow-up of at least 10 years. METHODS: Of the 1,211 patients who underwent TURP, 577 (47.6%) were available for follow-up. Long-term complications as well as reoperation rates were evaluated on the basis of a patient questionnaire or a physician interview. RESULTS: Repeat operation had to be performed in 35 patients (6%). TUR for bladder neck contracture had to be done in 14 patients (2.4%), while repeat TURP for BPH obstruction was required in 11 patients (1.9%). Finally, urethral strictures mandating surgical correction were present in 10 patients (1.7%). CONCLUSIONS: TURP is associated with low reoperation rates and therefore remains the gold standard in BPH therapy. Nevertheless, follow-up of these patients must be continued, as late complications requiring intervention may occur even after 10 years postoperatively. PMID- 14743464 TI - Phase I/II clinical trials of carbon ion therapy for prostate cancer. AB - BACKGROUND: Heavy ion beams possess high linear energy transfer components and a prominent Bragg peak in the human body, resulting in higher relative biological effectiveness and improved dose distribution. To establish heavy ion therapy techniques for the treatment of prostate cancer, phase I/II clinical trials were initiated. METHODS: For 96 patients with T1b-T3 prostate cancer, three carbon ion beams were used to irradiate the prostate and seminal vesicles (20 times/5 weeks) with or without endocrine therapy. Radiation dose was expressed in GyE which was initially thought to be equivalent to photon dose. Total dose was gradually increased from 54 to 72 GyE. RESULTS: Carbon ion therapy was completed in 20 cases of T1b/T1c/T2aN0M0 as monotherapy, in 8 cases of T2b/T3pN0M0 with neoadjuvant endocrine therapy, and in 68 cases of T2b/T3N0/pN1M0 with neoadjuvant and adjuvant endocrine therapy. Median observation period was 47 months. Grade 3 late radiation morbidity of rectum and/or bladder/urethra developed in one and five cases who received 66 and 72 GyE of radiation, respectively. After these adverse effects were observed, total dose was decreased to 66 GyE and the radiation field was coned down during the treatment course. At 5 years, overall, cause-specific, clinical recurrence-free, and biochemical recurrence-free survival rates were 87.7, 94.9, 90.0, and 82.6%, respectively. Local control was achieved in all patients except one patient who received 54 GyE of radiation. CONCLUSIONS: The therapeutic techniques of carbon ion therapy have been established for patients with prostate cancer. Carbon ion therapy may exert excellent effect to the tissues of prostate cancer. PMID- 14743465 TI - Long-term androgen-ablation causes increased resistance to PI3K/Akt pathway inhibition in prostate cancer cells. AB - BACKGROUND: In advanced stages of prostate cancer, the phosphatidylinositol-3' kinase (PI3K)/Akt signaling cascade, one of the major survival pathways in the cell, is frequently constitutively activated due to mutation or loss of the tumor suppressor protein phosphatase and tensin homolog deleted on chromosome 10 (PTEN). Using cell culture models representing different tumor stages, we explored the effect of inhibition of this survival pathway on the induction of apoptosis. METHODS: Inhibition of the survival kinase Akt and induction of apoptosis was analyzed in androgen-insensitive DU145 and PC-3 cells, in androgen responsive LNCaP, and in androgen-independent long-term androgen-ablated LNCaP abl cells representing therapy-resistant prostate cancer cells. Activated Akt was determined by immunoblotting using a phospho-Akt specific antibody. Induction of apoptosis was analyzed employing annexing V and propidium iodide staining and flow cytometry and measurement of cleavage of the caspases substrate poly-ADP ribose polymerase (PARP). RESULTS: IGF-1, EGF, and heregulin but not PDGF or activators of protein kinase A induced phosphorylation of Akt in DU145 cells and activation was completely blocked by the PI3K inhibitor LY294002. In the hormone responsive prostate cancer cell line LNCaP that has a constitutively switched-on Akt kinase, LY294002 caused a dose- and time-dependent Akt inhibition, which was absent in long-term androgen-ablated LNCaP sublines. In agreement with the resistance to inhibition of the PI3K/Akt pathway, long-term androgen-ablated LNCaP sublines remained relatively resistant to induction of cell death by LY294002 or the cytotoxic drug etoposide. Inhibition of the PI3K/Akt pathway restored the sensitivity of long-term androgen-ablated cells to induction of apoptosis by a cytotoxic drug almost completely. CONCLUSION: These results suggest that long-term androgen ablation therapy for prostate cancer reinforces the PI3K/Akt pathway and impedes its inhibition thus contributing to increased resistance of tumor cells to induction of apoptosis. With regard to treatment of therapy-refractory prostate cancer, these findings suggest effectiveness of a combination of cytotoxic treatment and inhibition of the PI3K-Akt survival pathway in tumor cells after failure of androgen-ablation therapy. PMID- 14743466 TI - Expression of p21 cell cycle protein is an independent predictor of response to salvage radiotherapy after radical prostatectomy. AB - BACKGROUND: To assess whether the expression of p21, p27, and p53 could predict biochemical failure in prostate cancer patients treated with neoadjuvant androgen deprivation prior to salvage radiotherapy for a rising post-radical prostatectomy (RP) prostate-specific antigen (PSA). METHODS: The expression of p21, p27, and p53 was determined by immunohistochemistry in a cohort of 74 formalin-fixed paraffin-embedded prostate cancer samples obtained from RP. Expression of these markers was then correlated with clinicopathological parameters and biochemical failure-free survival after salvage radiotherapy. RESULTS: Expression of p21, p27, and p53 was observed in 20%, 69%, and 74% of prostate cancer specimens, respectively. Overexpression of p21 correlated with a higher Gleason score (>7) (P = 0.024). Of the three markers, only p21 expression was correlated with PSA failure after radiotherapy (P = 0.034). In multivariate analysis, both positive p21 (P = 0.004) and pre-radiation serum PSA > 1 ng/ml (P < 0.0001) were independent predictors of biochemical failure after salvage radiotherapy. Patients with p21- tumors and a serum PSA level < or = 1 ng/ml before salvage radiotherapy had a biochemical failure-free survival at 5 years of 83%, compared to 16% at 5 years for those patients with either p21+ tumor or a PSA > 1 ng/ml. Patients with both p21+ and a PSA level > 1 ng/ml had a much lower biochemical failure-free survival rate of 25% at only 18 months (P < 0.0001). CONCLUSIONS: The expression of p21 in prostatectomy specimens could help predict the likelihood of response to salvage radiotherapy, particularly in patients treated before PSA reaches 1 ng/ml. PMID- 14743467 TI - Clinical and pathologic features of prostate cancer detected after repeat false negative biopsy in a screening population. AB - BACKGROUND: The present study was designed to investigate whether the clinical or pathologic features of prostate cancer (PCa) are related to the number of repeat biopsies required to establish the diagnosis of PCa. METHODS: Between February 1993 and August 2000, 653 patients were evaluated in this retrospective study. All patients underwent transrectal ultrasound-guided biopsy of the prostate prior to radical retropubic prostatectomy. The pathologic findings of specimens obtained at radical prostatectomy and pelvic lymph node dissection as well as PSA levels, findings on DRE, prostate volumes, transition zone volumes, and age were analyzed separately for all PCa patients diagnosed at the first set of biopsies (group A) and compared with the data of those diagnosed at the 2nd-5th set of biopsies (group B). In a second step, we compared the results obtained from patients diagnosed at the 2nd set of biopsies (group B1) with those of patients diagnosed at the 3rd to 5th set of biopsies (group B2). RESULTS: Gleason scores, pathologic tumor stages, and tumor volumes in group B were found to be significantly decreased compared to group A. But from the 2nd to 5th serial biopsy no further decrease in pathologic stage, Gleason score, or tumor volume was observed. On the contrary, there was a tendency towards higher tumor stages and Gleason scores. Of the tumors detected after the second false-negative set of biopsies almost 70% were lesions with Gleason scores of 6 or higher. CONCLUSIONS: False-negative results at the first needle biopsy are predictive of a lower pathologic stage and grade as well as smaller tumor volumes of PCa diagnosed at repeat sets of biopsies. False-negative results on repeat biopsy, however, have no prognostic significance for the tumor stage of PCas detected at subsequent sets of biopsies. PMID- 14743468 TI - In situ shortening of CAG repeat length within the androgen receptor gene in prostatic cancer and its possible precursors. AB - BACKGROUND: The amino-terminal transcriptional activation domain of the androgen receptor (AR) gene contains two polymorphic trinucleotide repeat segments that encode polyglutamine (CAG)n and polyglycine (GGC)n tracts. Shorter CAG repeat lengths are associated with higher transcriptional activity. The previous studies using peripheral blood leukocytes showed the relationship between shorter CAG repeat length and risk for prostate cancer (PCA). METHODS: Prostatic cancer (PCA), its possible precursors [high grade prostatic intraepithelial neoplasia (HGPIN) and postatrophic hyperplasia (PAH)], and non-neoplastic epithelium were microdissected from a whole-mount prostatectomy specimen from 34 cases with PCA. DNA extracted from each lesion was processed for PCR-based electrophoresis on 6% denaturing polyacrylamide gels, followed by direct sequencing. To examine whether the in situ shortening of CAG repeat was confined to the CAG repeat or was a part of phenomenon induced by microsatelite instability (MSI), BAT-25 and BAT-26, effective markers for detection of MSI, were also examined. RESULTS: All non neoplastic epithelial lesions had identical numbers of CAG repeat in the same prostate. CAG repeat lengths were identical in lesions in 25 cases. Two distinct products were found in 9 of 34 cases (26.5%); one product identical to that of non-neoplastic epithelium and another smaller one. In situ shortening of CAG repeat lengths in PCA, HGPIN, and PAH was found in 3 of 34 (8.8%), 6 of 34 (17.6%), and 3 of 10 (30%) cases, respectively. Frequency of CAG shortening was significantly higher in PAH than in PCA lesions (P < 0.05). The length of GGC repeats, BAT-25 and -26 was identical among all lesions in the same case. There was no significant correlation between shortening of CAG repeat length and the clinicopathologic parameters. CONCLUSION: Shortening of CAG repeat length was found in in situ lesions of PCA and its possible precursors. PMID- 14743469 TI - Anti-tumor effect of combination therapy with intratumoral controlled-release paclitaxel (PACLIMER microspheres) and radiation. AB - BACKGROUND: Paclitaxel is one of the few chemotherapeutics effective in patients with advanced protstate cancer. Paclitaxel has also been reported to have radiosensitizing effects in prostate cancer. Local delivery of a controlled release paclitaxel product may allow for increase local concentrations of paclitaxel at the tumor site and, in conjunction with radiation, may enhance cell kill by its radiosensitization mechanism. METHODS: Orthotopic LNCaP tumors were injected with 40% PACLIMER Microspheres (40% loading; w:w) when tumors were 100 200 mm(3). Twenty-eight days post cell injection, mice were sacrificed, tumors weighed, and serum measured for PSA. TSU-xenografts were injected with PACLIMER Microspheres (10% and 40% loaded; w:w) or placebo microspheres when the tumors were approximately 100 mm(3). Half of xenograft tumors were irradiated with a single dose (10 Gy) of radiation. Tumor volume was followed over time. RESULTS: Forty percent PACLIMER Microspheres significantly reduced tumor growth in the LNCaP orthotopic model. PSA was a good indicator of response. Forty percent PACLIMER Microspheres had a significant effect on slowing TSU growth compared to placebo microspheres. Addition of a single acute dose of radiation significantly enhanced the effect of 10% PACLIMER Microspheres (P < 0.05), had minimal effect on 40% PACLIMER Microspheres, and no enhancing effect on tumors treated with placebo microspheres. CONCLUSIONS: A controlled-release formulation of paclitaxel can be very effective in the treatment of prostate cancer. Additionally, PACLIMER Microspheres may be effectively used as a radiosensitizer in genitourinary cancers. PMID- 14743470 TI - Regulation of rat prostate stromal cell myodifferentiation by androgen and TGF beta1. AB - BACKGROUND: Myodifferentiation of stromal cells is a key step in prostate development and is a hallmark of reactive stroma in prostate cancer. Little is known about regulatory mechanisms, however, prostate stromal cells are androgen regulated and TGF-beta1 is a known stimulator of stromal myodifferentiation. The PS-1 rat prostate stromal cell line expresses androgen receptor, and exhibits androgen-regulated gene expression and proliferation. TGF-beta1 inhibits androgen action in PS-1 cells through translocation of androgen receptor from the nucleus to the cytoplasm. The present study was conducted to determine whether myodifferentiation of PS-1 cells is regulated by androgen and TGF-beta1, and how myodifferentiation affects androgen receptor localization and cell proliferation. METHODS: PS-1 cell cultures were exposed to physiological concentrations of dihydrotestosterone, TGF-beta1, and combinations of both in chemically defined medium. Immunocytochemistry and Western blotting for smooth muscle alpha-actin filament formation, smooth muscle alpha-actin protein levels, calponin expression, PCNA index, and androgen receptor localization were performed. RESULTS: Dihydrotestosterone (DHT) and TGF-beta1 each separately promoted PS-1 myodifferentiation. A combination did not affect the rate of differentiation, however, the level of alpha-actin protein was elevated and PCNA was decreased in co-stimulated conditions. TGF-beta1 induction resulted in a transient translocation of androgen receptor from the nucleus to the cytoplasm during differentiation followed by a resumed nuclear localization in myodifferentiated cells. CONCLUSIONS: These data indicate that a complex cross-talk mechanism exists between androgen and TGF-beta1 signaling in prostate stromal cells that affects cell proliferation and myodifferentiation. These findings also suggest that androgen and TGF-beta1 interactions may cooperatively regulate myodifferentiation of stromal cells in the stromal response in prostate cancer. PMID- 14743471 TI - Expression of NF-kappaB in prostate cancer lymph node metastases. AB - INTRODUCTION: Nuclear factor-kappaB (NF-kappaB) is a transcription factor that transactivates genes involved in the regulation of cell growth, apoptosis, angiogenesis, and metastasis. Our aim was to assess NF-kappaB expression in lymph node (LN) metastases of prostate cancer. METHODS: Immunohistochemical staining was performed using the p65 anti-NF-kappaB antibody. Seventy-seven paraffin embedded LN specimens obtained from 54 prostate cancer patients were analyzed. Of the 54 patients, 32 had positive LN metastases, while 22 showed no evidence of metastasis and were considered as controls. The overall percentage of NF-kappaB nuclear localization was assessed, as well as the intensity of staining. RESULTS: Nuclear localization of NF-kappaB was significantly greater in the metastatic LN group compared to controls. In patients with positive-LN metastases, 84.4% showed >10% nuclear staining in tumor cells. Moreover, 64.4% of the malignant LN specimens had >10% nuclear staining in lymphocytes compared to 0% in controls. Intensity of cytoplasmic and nuclear staining was higher in the metastatic LN group than in controls (P < 0.01). CONCLUSIONS: Nuclear localization/activation of NF-kappaB is up-regulated in prostate cancer LN metastasis. Such up-regulation of NF-kappaB activity is observed in the tumor cells as well as in the surrounding lymphocytes. PMID- 14743472 TI - Canine prostatic intraepithelial neoplasia: is the comparative model relevant? AB - BACKGROUND: Histologic sections from an archival collection of a veterinary teaching hospital were examined to determine the likelihood of detection of canine high-grade prostatic intraepithelial neoplasms (HGPIN), as a prelude to use of the canine model of prostatic carcinogenesis for chemopreventive strategies. METHODS: Tissue specimens representing clinically healthy (normal) prostate glands, benign prostatic hyperplasia, and prostatic carcinoma were examined in one tissue plane for histological evidence of HGPIN. RESULTS: No histological evidence of HGPIN was detected in 20 normal prostate glands or 95 prostate glands with benign prostatic hyperplasia. Seven of 20 prostatic carcinomas had synchronous HGPIN. CONCLUSIONS: Histological evidence of HGPIN is unlikely to be detected in the healthy or hyperplastic canine prostate gland with the clinically-procured biopsy. This might diminish the usefulness of canine HGPIN in temporal studies of chemoprevention of prostate cancer. HGPIN was found simultaneously with prostatic carcinoma in more than one-third of the carcinomas examined. PMID- 14743473 TI - Zeta potential of microfluidic substrates: 1. Theory, experimental techniques, and effects on separations. AB - This paper summarizes theory, experimental techniques, and the reported data pertaining to the zeta potential of silica and silicon with attention to use as microfluidic substrate materials, particularly for microchip chemical separations. Dependence on cation concentration, buffer and cation type, pH, cation valency, and temperature are discussed. The Debye-Huckel limit, which is often correctly treated as a good approximation for describing the ion concentration in the double layer, can lead to serious errors if it is extended to predict the dependence of zeta potential on the counterion concentration. For indifferent univalent electrolytes (e.g., sodium and potassium), two simple scalings for the dependence of zeta potential on counterion concentration can be derived in high- and low-zeta limits of the nonlinear Poisson-Boltzman equation solution in the double layer. It is shown that for most situations relevant to microchip separations, the high-zeta limit is most applicable, leading to the conclusion that the zeta potential on silica substrates is approximately proportional to the logarithm of the molar counterion concentration. The zeta vs. pH dependence measurements from several experiments are compared by normalizing the zeta based on concentration. PMID- 14743474 TI - Zeta potential of microfluidic substrates: 2. Data for polymers. AB - Zeta potential data are reviewed for a variety of polymeric microfluidic substrate materials. Many of these materials currently used for microchip fabrication have only recently been employed for generation of electroosmotic flow. Despite their recent history, polymeric microfluidic substrates are currently used extensively for microchip separations and other techniques, and understanding of the surface zeta potential is crucial for experimental design. This paper proposes the use of pC (the negative logarithm of the counterion concentration) as a useful normalization for the zeta potential on polymer substrates in contact with indifferent univalent counterions. Normalizing zeta by pC facilitates comparison of results from many investigators. The sparseness of available data for polymeric substrates prevents complete and rigorous justification for this normalization; however, it is consistent with double layer and adsorption theory. For buffers with indifferent univalent cations, normalization with the logarithm of the counterion concentration in general collapses data onto a single zeta/pC vs. pH curve, and (with the exception of PMMA) the repeatability of the data is quite encouraging. Normalization techniques should allow improved ability to predict zeta potential performance on microfluidic substrates and compare results observed with different parameters. PMID- 14743475 TI - Fluid mechanics of electroosmotic flow and its effect on band broadening in capillary electrophoresis. AB - Electroosmotic flow (EOF) usually accompanies electrophoretic migration of charged species in capillary electrophoresis unless special precautions are taken to suppress it. The presence of the EOF provides certain advantages in separations. It is an alternative to mechanical pumps, which are inefficient and difficult to build at small scales, for transporting reagents and analytes on microfluidic chips. The downside is that any imperfection that distorts the EOF profile reduces the separation efficiency. In this paper, the basic facts about EOF are reviewed from the perspective of fluid mechanics and its effect on separations in free solution capillary zone electrophoresis is discussed in the light of recent advances. PMID- 14743476 TI - Sample introduction techniques for microfabricated separation devices. AB - A great deal of progress has been made toward the development of the micro total analysis system (micro-TAS) since its inception in 1990. A wide variety of applications, including genomics, proteomics and drug discovery, have prompted the development of analytical methods capable of very high throughput while maintaining low cost. The micro-TAS concept addresses both of these requirements. Electrophoresis has been a key element in the development of the micro-TAS. Most chemical and biochemical assays utilize a separation component at some point during analysis. Genomics, in particular, depends almost exclusively on electrophoresis for size-based separations of DNA. This review examines sample introduction into microfabricated electrophoretic devices, or chips, primarily for DNA analysis. Sample introduction is an important component of these systems and is an essential process for making chip electrophoresis a widely applicable analytical technique. Specific issues, such as automation, the delivery of large numbers of samples to microfabricated devices and injection of picoliter-sized sample plugs into a separation lanes on chips, are presented. PMID- 14743477 TI - Cyclic electrophoretic and chromatographic separation methods. AB - A review is given of the application of cyclic analytical methods in capillary electroseparation (CE) and liquid chromatography (LC) systems. Cyclic methods have been used since the early sixties in chromatographic systems to overcome pressure limitations to resolution. From the early nineties on they have also been applied in capillary electroseparation systems to overcome voltage limitations. Some basic theory is given, outlining the temporal development of resolution in cyclic CE and LC systems and calculating the maximal resolution that can be obtained as a function of the operational parameters of pressure and electrical field. Simple equations are given for the temporal change in the peak capacity and the loss of peaks from the systems as it occurs in some cyclic systems. Finally, a circular open tubular chromatographic system is proposed using integrated pumping and continuous detection. The performance of such a system is discussed using magnetohydrodynamic and alternating current electroosmotic pumping as examples of integrated pumps and Shah Convolution Fourier transform detection as an example of a continuous detection method. PMID- 14743478 TI - Effect of Joule heating on efficiency and performance for microchip-based and capillary-based electrophoretic separation systems: a closer look. AB - An attempt is made to revisit the main theoretical considerations concerning temperature effects ("Joule heating") in electro-driven separation systems, in particular lab-on-a-chip systems. Measurements of efficiencies in microfabricated devices under different Joule heating conditions are evaluated and compared to both theoretical models and measurements performed on conventional capillary systems. The widely accepted notion that planar microdevices are less susceptible to Joule heating effects is largely confirmed. The heat dissipation from a nonthermostatically controlled glass microdevice was found to be comparable to that from a liquid-cooled-fused silica capillary. Using typically dimensioned glass and glass/silicon microdevices, the experimental results indicate that 5-10 times higher electric field strengths can be applied than on conventional capillaries, before detrimental effects on the separation efficiency occur. The main influence of Joule heating on efficiency is via the establishment of a radial temperature profile across the lumen of the capillary or channel. An overall temperature increase of the buffer solution has only little influence on the quality of the separation. Still, active temperature control (cooling, thermostatting) can help prevent boiling of the buffer and increase the reproducibility of the results. PMID- 14743479 TI - Optimisation of selectivity in the separation of aromatic amino acid enantiomers using sulfated beta-cyclodextrin and dextran sulfate as pseudostationary phases. AB - Control of selectivity in the enantiomeric separation of three aromatic amino acids (phenylalanine, tyrosine and tryptophan) was demonstrated utilising two separate electrolyte additives. Sulfated-beta-cyclodextrin (s-beta-CD) was chosen as the chiral selector while the addition of dextran sulfate provided a means with which to predictably fine-tune separation selectivity. The two additives were found to interact independently with the amino acids, with the s-CD providing chiral interactions while the dextran sulfate provided ion-exchange (IE) interactions. The system was also very robust with reproducibility of migration times being < 2.0% RSD between runs and < 2.6% on using a new capillary. A physical model derived from first principles was also successfully used to describe the two additive system. The model accurately described the observed separations over the range of 0-20 mM s-beta-CD and 0-1% dextran sulfate with a correlation coefficient of 0.998 between predicted and observed mobilities. The physical model also provided useful information about the system including association constants between the analytes and the pseudostationary phases, together with the mobilities of the associated complexes (analyte cyclodextrin and analyte-dextran sulfate). Selectivity optimisation was achieved using the normalised resolution product and minimum resolution criteria. The physical model also allowed a desired separation selectivity to be obtained, such that experimental conditions could be predicted to lead to a particular migration order. PMID- 14743480 TI - Novel enantioselective strong cation exchangers based on sulfodipeptide selectors: evaluation for enantiomer separation of chiral bases by nonaqueous capillary electrochromatography. AB - Strong cation exchange (SCX)-type chiral stationary phases (CSPs) based on beta amino sulfonic acid-terminated dipeptide derivatives as chiral selectors, immobilized on thiol-modified silica particles (3.5 microm), were synthesized and applied to enantiomer separations of chiral bases by nonaqueous capillary electrochromatography (CEC). The effect of structural variations of the sulfodipeptide selectors on the separation factors alpha was investigated. These studies included variation of the acid-terminal amino sulfonic acid residue, variation of the configurations, i.e., comparison of the diastereomeric (S,S)- and (R,S)-configurations of the sulfodipeptides, and finally comparison of sulfodipeptide selectors with corresponding beta-amino sulfonic acid analogs. In general, the capillary columns (100 microm ID) packed with the new SCX-type CSPs showed enantioselectivity for an elaborated set of chiral basic drugs in CEC acting by an enantioselective cation-exchange retention mechanism. N-[N-(4 Allyloxy-3,5-dichlorobenzoyl)-leucyl]-2-amino-3,3-dimethylbutane sulfonic acid, in particular with (R,S)-configuration, turned out to be a more effective SCX type selector than a more rigid analog based on N-[N-(4-Allyloxy-3,5 dichlorobenzoyl)-leucyl]-2-pyrrolidinemethane sulfonic acid. Both of the former diastereomers were capable to baseline-resolve the enantiomers of ca. 40% of the tested basic chiral solutes including sympathomimetics and beta-blockers, while for the latter SCX-type CSPs only 10-20% of the selected solutes afforded resolutions > 1.5. PMID- 14743481 TI - Study of enzymatic reaction by electrophoretically mediated microanalysis in a partially filled capillary with indirect or direct detection. AB - Electrophoretically mediated microanalysis (EMMA), in combination with a partial filling technique and indirect or direct detection, is described for the study of enzymes reacting with the high mobility inorganic or organic anions as substrates or products. Part of the capillary is filled with a buffer optimized for the enzymatic reaction, the rest of the capillary with the background electrolyte being optimal for the separation of substrates and products. With haloalkane dehalogenase, chosen as a model enzyme, the enzymatic reaction was performed in a 20 mM glycine buffer (pH 8.6). Because of the wide substrate specificity of this enzyme, utilizing chlorinated as well as brominated substrates and producing either nonabsorbing chloride or absorbing bromide ions, two different background electrolytes and detection approaches were adopted. A 10 mM chromate-0.1 mM cetyltrimethylammonium bromide background electrolyte (pH 9.2) was used in combination with indirect detection and 20 mM beta-alanine-hydrochloric acid (pH 3.5) in combination with direct detection. The Michaelis constant (K(m)) of haloalkane dehalogenase for 1-bromobutane was determined. The K(m) values 0.59 mM estimated by means of indirect detection method and 0.17 mM by means of direct detection method were comparable with the value 0.13 mM estimated previously by gas chromatography. PMID- 14743482 TI - Ongoing development of two-dimensional polyacrylamide gel electrophoresis data standards. AB - We present an approach toward standardizing two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) data in support of developing a globally relevant proteomics consensus in order to provide more efficient database querying and data comparisons through the establishment of the necessary definitions and interdisciplinary reference fields for both the 2-D PAGE community, particularly in the proteomics area, and the clinical and experimental biological research communities, in general. This article covers the need for unifying the 2-D PAGE data through a common data repository, and its usefulness in data standards and data interoperability. PMID- 14743483 TI - Determination of cationic mobilities and pKa values of 22 amino acids by capillary zone electrophoresis. AB - The effective mobilities of the cationic forms of common amino acids--mostly proteinogenic--were determined by capillary zone electrophoresis in acidic background electrolytes at pH between 2.0 and 3.2. The underivatized amino acids were detected by the double contactless conductivity detector. Experimentally measured effective mobilities were fitted with the suitable regression functions in dependence on pH of the background electrolyte. The parameters of the given regression function corresponded to the values of the actual mobilities and the mixed dissociation constants (combining activities and concentrations) of the compound related to the actual ionic strength. McInnes approximation and Onsager theory were used to obtain thermodynamic dissociation constants (pK(a)) and limiting (absolute) ionic mobilities. PMID- 14743484 TI - Determination of the cis-trans isomerization barrier of enalaprilat by dynamic capillary electrophoresis and computer simulation. AB - Dynamic capillary electrophoresis (DCE) and computer simulation of the elution profiles with the stochastic model has been applied to determine the isomerization barriers of the angiotensin converting enzyme inhibitor enalaprilat. The separation of the rotational cis-trans isomeric drug has been performed in an aqueous 20 mM borate buffer at pH 9.3. Interconversion profiles featuring plateau formation and peak broadening were observed. To evaluate the rate constants k(cis-->trans) and k(trans-->cis) of the cis-trans isomerization from the experimental electropherograms obtained by dynamic capillary electrophoresis, elution profiles were analyzed by a simulation with iterative convergence to the experimental data using the ChromWin software which requires the total migration times of the individual isomers t(R), the electroosmotic break-through time t(0), the plateau height h(plateau), the peak widths at half height of the individual isomers w(h), as well as the peak ratio of the isomers as experimental data input. From temperature-dependent measurements between 0 degrees and 15 degrees C the thermodynamic parameters Delta G, Delta H and Delta S, the rate constants k(cis-->trans) and k(trans-->cis) and the kinetic activation parameters Delta G*, Delta H*, and Delta S* of the cis-trans isomerization of enalaprilat were obtained. From the activation parameters the isomerization barriers at 37 degrees C were calculated to be Delta G* (trans- >cis) = 87.2 kJ.mol(-1) and Delta G*(cis-->trans) = 91.9 kJ.mol(-1). PMID- 14743485 TI - High-resolution computer simulation of the dynamics of isoelectric focusing of proteins. AB - A dynamic electrophoresis simulator that accepts 150 components and voltage gradients employed in the laboratory was used to provide a detailed description of the focusing process of proteins under conditions that were hitherto inaccessible. High-resolution focusing data of four hemoglobin variants in a convection-free medium are presented for pH 3-10 and pH 5-8 gradients formed with 20 and 40 carrier ampholytes/pH unit, respectively. With 300 V/cm, focusing is shown to occur within 5-10 min, whereas at 600 V/cm separation is predicted to be complete between 2.5 and 5 min. The time interval required for focusing of proteins is demonstrated to be dependent on the input protein charge data and, however less, on the properties of the carrier ampholytes. The simulation data reveal that the number of transient protein boundaries migrating from the two ends of the column towards the focusing positions is equal to the number of sample components. Each protein is being focused via the well-known double-peak approach to equilibrium, a process that is also characteristic for focusing of the carrier ampholytes. The predicted focusing dynamics for the hemoglobin variants in pH 3-10 and pH 5-8 gradients are shown to qualitatively agree well with experimental data obtained by whole-column optical imaging. PMID- 14743486 TI - Calibrationless quantitative analysis by indirect UV absorbance detection in capillary zone electrophoresis: the concept of the conversion factor. AB - A new, fast and efficient procedure is described for the simultaneous quantitative analysis of various non-UV absorbing species in a sample, by capillary zone electrophoresis with indirect UV absorbance detection. The procedure is based on the concept of the conversion factor (CF). The CF of an analyte is defined as the ratio of the measured temporal peak area and the product of its migration time and transfer ratio (TR). Thus defined, the CF is of general validity for all analytes separated and detected in a given background electrolyte (BGE), since it has the same value for the same amounts of various analytes. If a sample is enriched with a known concentration of a standard component and analyzed by CZE, the CF of this standard component can be calculated and then the concentrations of all other analytes can be determined, without the use of any calibration graph. The individual TRs can be determined a priori from tabulated ionic mobilities and pK values of the analytes and of the constituents of the BGE or, for strong analytes, by using experimental data from the electropherogram of the analysis itself. The practical procedure of the analysis includes enrichment of the sample with a known quantity of a suitable standard and a single CZE run of the resulting mixture. The injected volume does not need to be known and thus the procedure also eliminates the injection error. The proposed procedure has been verified experimentally and reproducible and accurate values were obtained by using four different CZE apparatus for the analyses of standard mixtures of cations in three different BGEs. PMID- 14743487 TI - Peak dispersion and contributions to plate height in nonaqueous capillary electrophoresis at high electric field strengths: ethanol as background electrolyte solvent. AB - Nonaqueous capillary electrophoretic separations were performed under high electric field strengths (up to 2000 Vcm(-1)) in ethanolic background electrolyte solution and the contributions of different band broadening effects to plate height were evaluated. Under optimum conditions, increasing the field strength will provide faster separations and increased separation efficiency. Decrease in the separation efficiency at high field strengths was, however, observed in a previous study and now in the present paper an attempt is made to quantify various band broadening effects by applying a plate height model, which included the contributions of the injection plug length, diffusion, electromigration dispersion, Joule heating, analyte adsorption to the capillary wall, and detector slit aperture length. Of special interest were the contributions of Joule heating and analyte adsorption to the capillary wall. Poly(glycidylmethacrylate-co-N vinylpyrrolidone)-coated fused-silica capillaries were used with internal diameters (ID) ranging from 30 to 75 microm. The separation efficiencies obtained experimentally were compared with the theoretically calculated efficiencies and fairly good agreement was observed for the 30 microm ID capillary. Relatively large deviation from the predictions of the model was found for the other capillary diameters especially at higher field strengths. The possible reasons for the deviation were discussed. PMID- 14743488 TI - Experimental assessment of electromigration properties of background electrolytes in capillary zone electrophoresis. AB - Electromigration dispersion (EMD) properties of background electrolytes (BGEs) used in capillary zone electrophoresis (CZE) are of key importance for the success of an analysis. The knowledge of these properties may serve well for the prediction of the asymmetry of peaks of analytes, for the prediction of unsafe regions where a strong interference of system zones may be expected, and for the selection of optimum conditions where the analytes of interest may give sharp and practically symmetric peaks. Present theories enable one to calculate and predict EMD properties of many BGEs but there is also a lot of BGEs that are beyond the present theoretical models as far as their composition and equilibria involved are considered. This contribution brings a method for assessment of EMD properties of any BGE from easily accessible experimental data. The method proposed is illustrated by model examples both for cationic and anionic separations. Imidazole acetate, histamine acetate, and histidine acetate served as model BGEs for cationic separations; as the model BGE for anionic separations, Tris-borate and sodium-borate BGEs have been selected since these buffers are frequently used and borate is well-known for its complexing equilibria in aqueous solutions. PMID- 14743489 TI - Preparative free-flow electrofocusing in a vortex-stabilized annulus. AB - This paper describes the development of an annular chamber designed for preparative free-flow electrophoresis which can operate at voltages up to 20 kV, electric fields up to 65 kV/m, and power densities as high as 10 MW/m(3). This apparatus uses counter-rotating fluid vortices to stabilize the aqueous medium in the annulus against natural convection while improving heat and mass transfer over vortex-free Couette flow. The vortices are generated by rotating the inner surface of the annulus and, if the rotor and stator have complementary shapes, dispersion induced by electrokinetic and electrohydrodynamic flows can be mitigated even at the highest operating voltages. Following a brief overview of contemporary annular free-flow instrumentation, the theoretical principles for momentum and heat transport in the vortices are discussed in some detail and then the results of several electrofocusing experiments are provided to illustrate the resolving power of this instrument. PMID- 14743490 TI - Nafion membrane electrophoresis with direct and simplified end-column pulse electrochemical detection of amino acids. AB - A novel electrophoresis technique, in which the separation column was replaced by a strip of Nafion membrane (5.0 cm x 0.20 mm x 0.25 mm), was developed for the separation of an amino acid mixture (glycine, asparic acid and lysine), followed by quadruple-pulse electrochemical detection. Nafion membrane contains hydrophilic pores (10-20 A and 50-60 A in size) acting as very narrow electrophoresis channels. The fixed-charge sites (-SO(3) (-)) on the hydrophilic pore surface provide a strong charged background. A platinum disk electrode (0.90 mm inner diameter) was employed as the detection electrode and the electrophoresis cathode was used as the quasi-reference and counter electrode for the end-column electrochemical detector, without decoupler. Under optimized conditions the mixture of amino acids could be separated at a voltage of only 90 V with a detection limit of 10(-7) M, indicating that Nafion membrane electrophoresis is a potentially attractive technique for the separation of small organic molecules or ions. PMID- 14743491 TI - Preparative-scale, recirculating, pH-biased binary isoelectric trapping separations. AB - In order to improve the production rates and lower the specific electrophoretic energy consumption values in preparative-scale, recirculating, binary isoelectric trapping separations, we propose to add an auxiliary isoelectric agent to the solution in the anodic separation compartment and another to the solution in the cathodic separation compartment to implement pH-biased isoelectric trapping. The auxiliary isoelectric agents are selected such that they are trapped in the respective anodic and cathodic separation compartments and also, have isoelectric point (pI) values that are different from the pI values of the analytes of interest. By proper selection of the auxiliary isoelectric agents and their concentrations, the analytes of interest can be kept in nonisoelectric, charged state during the entire course of the preparative-scale, recirculating, binary isoelectric trapping separation. This results in higher electrophoretic mobilities and solubilities for the analytes than in their isoelectric or near isoelectric states, and leads to faster binary isoelectric trapping separations. PMID- 14743495 TI - Emphysema: the challenge of the remodelled lung. AB - Emphysema is recognized as the component of chronic obstructive airways disease that is responsible for airways obstruction. Different patterns of emphysema are, however, recognized, suggesting possible different pathogenetic processes within the lung. This, coupled with the associated idea of susceptibility factors to the development of emphysema, has led to studies of genes that may be involved in the defence of the lung from proteolytic and oxidative damage. These studies have been driven by the goal of finding a treatment for emphysema, but appear to have lost sight of the fundamental remodelling of the lung that has occurred in patients with emphysema and the fact that it is not a single morphological entity. PMID- 14743496 TI - Role of inflammatory mediators in the pathophysiology of acute respiratory distress syndrome. AB - Inflammatory response leading to organ dysfunction and failure continues to be the major problem after injury in many clinical conditions such as sepsis, severe burns, acute pancreatitis, haemorrhagic shock, and trauma. In general terms, systemic inflammatory response syndrome (SIRS) is an entirely normal response to injury. Systemic leukocyte activation, however, is a direct consequence of a SIRS and if excessive, can lead to distant organ damage and multiple organ dysfunction syndrome (MODS). When SIRS leads to MODS and organ failure, the mortality becomes high and can be more than 50%. Acute lung injury that clinically manifests as acute respiratory distress syndrome (ARDS) is a major component of MODS of various aetiologies. Inflammatory mediators play a key role in the pathogenesis of ARDS, which is the primary cause of death in these conditions. This review summarizes recent studies that demonstrate the critical role played by inflammatory mediators such as tumour necrosis factor (TNF)-alpha, interleukin (IL)-1beta, IL-6, platelet activating factor (PAF), IL-10, granulocyte macrophage colony stimulating factor (GM-CSF), C5a, intercellular adhesion molecule (ICAM) 1, substance P, chemokines, VEGF, IGF-I, KGF, reactive oxygen species (ROS), and reactive nitrogen species (RNS) in the pathogenesis of ARDS. It is reasonable to speculate that elucidation of the key mediators in ARDS coupled with the discovery of specific inhibitors would make it possible to develop clinically effective anti-inflammatory therapy. PMID- 14743497 TI - Tissue and cellular tropism of the coronavirus associated with severe acute respiratory syndrome: an in-situ hybridization study of fatal cases. AB - Severe acute respiratory syndrome (SARS) is a new human infectious disease with significant morbidity and mortality. The disease has been shown to be associated with a new coronavirus (SARS-CoV). The clinical and epidemiological aspects of SARS have been described. Moreover, the viral genome of SARS-CoV has been fully sequenced. However, much of the biological behaviour of the virus is not known and data on the tissue and cellular tropism of SARS-CoV are limited. In this study, six fatal cases of SARS were investigated for the tissue and cellular tropism of SARS-CoV using an in-situ hybridization (ISH) technique. Among all the tissues studied, positive signals were seen in pneumocytes in the lungs and surface enterocytes in the small bowel. Infected pneumocytes were further confirmed by immunofluorescence-fluorescence in-situ hybridization (FISH) analysis. These results provide important information concerning the tissue tropism of SARS-CoV, which is distinct from previously identified human coronaviruses, and suggest the possible involvement of novel receptors in this infection. Whereas the lung pathology was dominated by diffuse alveolar damage, the gut was relatively intact. These findings indicated that tissue responses to SARS-CoV infection are distinct in different organs. PMID- 14743498 TI - Expression of IL-27 in human Th1-associated granulomatous diseases. AB - Interleukin (IL)-27 is a newly described member of the IL-12 family. It is a heterodimeric cytokine composed of two subunits, p28 and Epstein-Barr virus induced gene 3 (EBI3). In vitro studies have shown that IL-27 is mainly produced by activated monocytes and dendritic cells. It induces the proliferation of naive CD4-positive T cells and synergizes with IL-12 for interferon-gamma (IFN-gamma) production. Knock-out mice for the IL-27 receptor (WSX-1/TCCR) have impaired Th1 responses and form abnormal granulomas when injected with bacillus Calmette Guerin. However, the expression profile of IL-27 in vivo is currently unknown. To investigate the potential role of IL-27 in the development of a Th1 response in humans in vivo, this study has analysed the in situ expression of IL-27 subunits in three types of granulomatous disease (tuberculosis, sarcoidosis, and Crohn's disease), each characterized by a Th1 response. Tissue sections from patients with tuberculosis (n = 9), sarcoidosis (n = 8), or Crohn's disease (n = 7) were analysed by immunohistochemistry with anti-EBI3 and anti-p28 antibodies, in parallel with control tissues (control reactive lymph nodes, n = 14, and control intestinal tissues, n = 11). In granulomatous tissues, EBI3 and p28 co-expression was detected in epithelioid and multinucleate giant cells in granulomas. In addition, sinus or tissue macrophages, endothelial cells, and plasma cells were found to co-express EBI3 and p28. These data support a possible role for IL-27 in human Th1 responses. PMID- 14743499 TI - The human Delta-like 1 homologue is implicated in the progression of liver fibrosis in biliary atresia. AB - Advanced liver cirrhosis frequently occurs in infants with biliary atresia despite early surgical correction. The aetiology is unknown, but may involve many cytokines and liver cells including hepatic stellate cells (HSCs). A cytokine expression array and real-time quantitative reverse transcription-polymerase chain reaction (QRT-PCR) were used to study cytokine expression during the progression of liver fibrosis in biliary atresia. A Delta-like 1 homologue (DLK1) gene was identified and this gene was up-regulated during the early stage, and down-regulated during the late stage, of biliary atresia, similar to the expression pattern of the procollagen alpha1(I) gene. Further characterization with immunohistochemistry, confocal microscopy, and in situ hybridization revealed that the DLK1 protein was mainly present in the cytoplasm of smooth muscle actin-positive mesenchymal cells that were morphologically and immunohistochemically identical to activated HSCs/myofibroblasts, whereas DLK1 mRNA was present only in hepatocytes. As DLK1 is a negative regulator of adipocyte differentiation and may control cell fate during differentiation, overexpression of DLK1 protein in HSCs in the early stage of biliary atresia suggests that DLK1 may be implicated in the transformation of HSCs from fat storing cells to myofibroblasts and in fibrogenesis associated with biliary atresia. PMID- 14743500 TI - A comparative study of mRNA and protein expression of the autoimmune regulator gene (Aire) in embryonic and adult murine tissues. AB - Autoimmune polyendocrine syndrome type 1 (APS1) is a rare autosomal recessive human disorder caused by mutations in the autoimmune regulator gene (AIRE) and characterized by multiple autoimmune diseases. As reports of the tissue expression pattern of the murine Aire gene are discordant, a comprehensive survey of Aire expression was undertaken in adult and embryonic tissues at the mRNA and protein levels using real-time RT-PCR, in situ hybridization, and immunohistochemistry. In the adult, the highest Aire mRNA expression was in the thymus. All the other tissues investigated expressed Aire mRNA at low levels, but it was barely detectable in the adrenal gland. Aire protein expression was observed in the thymus, spleen, and lymph nodes. A common pattern was observed in other tissues, with staining in epithelial cells. An exception to this was the gut, where staining was seen in the mucin spaces. In embryonic tissue, Aire mRNA and protein expression was detected from E14.5 in the thymus. In the fetal liver, unlike the adult, staining was observed at E14.5 and decreased towards term. Thus, Aire is expressed in immunologically relevant tissues and in a restricted number of extra-immunological tissues in the adult. Furthermore, the presence of Aire protein is reported in extra-thymic tissues of the embryo. PMID- 14743501 TI - Expression of acyl-CoA synthetase 5 reflects the state of villus architecture in human small intestine. AB - Several disorders of the small intestine are associated with disturbances in villus architecture. Thus, an understanding of the molecular mechanisms associated with the differentiation of villi represents an important step in the improvement of the understanding of small intestinal pathology. Screening of antibodies from a hybridoma library led to the identification of an acyl-CoA synthetase 5-specific monoclonal antibody. Protein synthesis, mRNA expression, and the enzyme activity of acyl-CoA synthetase 5 were studied by several methods in human small intestinal tissues with Crohn's disease or coeliac disease, respectively. Acyl-CoA synthetase 5 mRNA and protein levels were substantially reduced in injured small intestinal mucosa. Moreover, impaired synthesis of the acyl-CoA synthetase 5 protein was reflected by a decrease in intramucosal enzyme activity. Subtle changes of the acyl-CoA synthetase 5 pattern correlate with conversion of intestinal epithelial cells to a gastric phenotype. These results suggest that deranged acyl-CoA synthetase 5 expression, synthesis, and activity are closely related to the state of villus architecture and epithelial homeostasis in human small intestine. PMID- 14743502 TI - Gastric mucosal cytokine and epithelial cell responses to Helicobacter pylori infection in Mongolian gerbils. AB - Experimental infection with Helicobacter pylori in Mongolian gerbils results in chronic gastritis and gastric cancer. To investigate epithelial cell proliferation, apoptosis, and mucosal cytokine responses in gastritis, Mongolian gerbils were infected with the H pylori SS1 strain. At 4 weeks post-infection, gastritis was predominantly within the antrum, but extended to the corpus in approximately 50% of gerbils by 36 weeks. Epithelial cell proliferation and apoptosis in glandular epithelial cells were increased with infection. Antral cell proliferation, but not apoptosis, correlated significantly with gastric inflammation. In female gerbils, H pylori significantly increased expression of transcripts for IFN-gamma and IL-12p40, but not TGF-beta or IL-10, in the gastric mucosa. Significantly reduced IFN-gamma and IL-12p40 responses were observed in male gerbils infected with H pylori, but epithelial proliferative and apoptotic responses were comparable to those of females. These studies demonstrate that the female gerbil cytokine response to H pylori has a Th1 profile and that there are gender differences in the magnitude of the gastric cytokine responses to H pylori. The absence of a down-regulatory cytokine response may account for the more severe gastritis observed with H pylori infection in gerbils than in mice. PMID- 14743503 TI - Expression of BUB1 protein in gastric cancer correlates with the histological subtype, but not with DNA ploidy or microsatellite instability. AB - During mitosis, the spindle checkpoint delays the onset of anaphase until all chromosomes have attached properly to the mitotic spindle, preventing chromosome missegregation. BUB (budding uninhibited by benzimidazole) 1 is one of the key components of this checkpoint. BUB1 mutations are rare in cancer tissues and no mutations have been identified in gastric cancer. In mice, immunodepletion of BUB1 abolished the spindle checkpoint. Thus, aberrant expression of BUB1 protein could impair mitotic checkpoint function, resulting in aneuploidy, a common phenomenon in gastric cancer. In the present study, an antibody was generated against BUB1 and its expression was studied in gastric cancer tissue sections (n = 80) by immunohistochemistry. Nuclear BUB1 expression was found in all gastric cancer cases. The proportion of tumour cells expressing BUB1 was significantly greater in diffuse-type than in intestinal-type gastric carcinoma (p < 0.001). No correlation was found between BUB1 expression and deoxyribonucleic acid (DNA) ploidy, microsatellite instability or any other histopathological parameters investigated. To the authors' knowledge, this is the first study of BUB1 protein expression in gastric cancer tissues. Different BUB1 protein expression levels in intestinal- and diffuse-type gastric cancer may provide further evidence of a potential link between different genetic pathways and morphological phenotype in gastric carcinogenesis. However, further studies are needed to establish whether there is an association between BUB1 protein expression level and mitotic spindle checkpoint function in gastric cancer. PMID- 14743504 TI - Expression of BRCA1 protein in benign, borderline, and malignant epithelial ovarian neoplasms and its relationship to methylation and allelic loss of the BRCA1 gene. AB - BRCA1 is a putative tumour suppressor gene responsible for a hereditary ovarian cancer syndrome. To clarify the possible involvement of BRCA1 in the development of sporadic ovarian neoplasms, this study analysed the immunohistochemical expression of BRCA1 protein in normal ovarian surface epithelium and 119 epithelial ovarian tumours (19 benign, 24 borderline, and 76 malignant tumours). Loss of heterozygosity (LOH) of BRCA1 was examined using three microsatellite markers to analyse the relationship between BRCA1 expression and alterations of the BRCA1 gene. Methylation of the BRCA1 promoter was also analysed by methylation-specific PCR. In ovarian carcinomas showing heterogeneous expression of BRCA1 protein in the same tumour, LOH and methylation status were analysed using microdissection techniques. Finally, the relationship of BRCA1 expression or its genetic alteration to clinicopathological parameters and patient survival was analysed. Ovarian surface epithelial cells expressed BRCA1 protein. Decreased expression of BRCA1 was found in 16% of benign tumours, 38% of borderline tumours, and 72% of carcinomas. LOH of BRCA1 was demonstrated in no benign tumours, 15% of borderline tumours, and 66% of carcinomas. Methylation of BRCA1 was not detected in benign or borderline tumours, but was present in 31% of carcinomas. Reduced expression of BRCA1 correlated with the presence of gene methylation. The frequency of BRCA1 methylation and LOH was higher in serous carcinomas than in other types. In one of the three serous carcinomas that showed heterogeneous expression of BRCA1, BRCA1-positive borderline-like tumour cells were LOH-positive and methylation-negative, whereas adjacent BRCA1-negative carcinoma cells were LOH-positive and methylation-positive. The prognosis of carcinoma patients did not correlate with BRCA1 expression or genetic status. These findings suggest that reduced expression of BRCA1 protein along with genetic and epigenetic changes of the BRCA1 gene play an important role in the development of sporadic ovarian carcinomas, particularly those of serous histology. PMID- 14743505 TI - Altered expression of the ZBRK1 gene in human breast carcinomas. AB - The ZBRK1 protein is a member of the KRAB-ZFP family. It functions as a transcriptional repressor by binding to its recognition sequence within target genes and producing a nucleoprotein complex containing its co-repressor BRCA1 and also probably the co-repressor KAP-1. Alterations that affect the ZBRK1 gene have not been reported in human tumours. For this reason, possible alterations in the ZBRK1 gene have been studied by analysing mRNA expression using real-time polymerase chain reaction (PCR), and gene sequence using single strand conformation polymorphism (SSCP) analysis and DNA sequencing, in 61 patients with primary breast carcinomas. BRCA1 mRNA expression and allelic loss were also studied in 25 of the same patients. ZBRK1 was underexpressed in 28 (45.9%) and overexpressed in 18 (29.5%) of the 61 cases. No significant association was observed between ZBRK1 mRNA expression and clinicopathological parameters. Loss of heterozygosity of the BRCA1 gene was found in 3 of 23 (13%) informative cases and BRCA1 mRNA expression was altered in 11 of 25 (44%) cases. No significant correlation was found between altered BRCA1 expression and the different types of ZBRK1 mRNA expression. Nine polymorphisms were found in the ZBRK1 sequence, with no significant differences between patients and control subjects. Altered ZBRK1 expression correlated significantly with two of these polymorphisms. A point mutation in one polymorphism and allelic loss in one patient were also observed, findings that indicate that these inactivation mechanisms do not frequently affect this gene. Altered expression of the ZBRK1 gene is therefore frequent in primary breast carcinoma. The functional significance of the polymorphisms in this gene is unclear. PMID- 14743506 TI - Aberrant CpG island hypermethylation of multiple genes in prostate cancer and prostatic intraepithelial neoplasia. AB - To date, several reports have been published about CpG island methylation of various genes in prostate cancer. However, most of these studies have focused on cancer tissue only or a single gene and data about concurrent methylation of multiple genes in prostate cancer or prostatic intraepithelial neoplasia (PIN) are limited. The aim of the present study was to determine the methylation profile of 11 tumour-related genes in prostate cancer and PIN. Seventy-one samples, including 37 prostate cancers, 14 PINs, and 20 normal prostates, were examined for the methylation status of 11 tumour-related genes using methylation specific PCR. The mean number of genes methylated was significantly higher in prostate cancer and PIN than in non-neoplastic prostate (4.4, 3, and 0.2, respectively; p < 0.001). In prostate cancer, APC, GSTP1, MGMT, and RASSF1A were frequently methylated at a frequency of 56.8%, 86.5%, 75.7%, and 83.8%, respectively. These genes were methylated in more than 30% of PINs. Prostate cancers with high serum prostate-specific antigen (PSA) (more than 8 ng/ml) or a high Gleason score (GS) (3 + 4 or more) showed higher numbers of methylated genes than those with low serum PSA (8 or less) or low GS (3 + 3 or less) (5.4 versus 2.5 and 5.4 versus 3.1, respectively; p < 0.05). The methylation frequency of APC, RASSF1A, and RUNX3 was higher in prostate cancers with high serum PSA or with high GS than in those with low PSA or with low GS, respectively, the differences reaching statistical significance (p < 0.05). A strong association between MGMT methylation and loss of MGMT expression was demonstrated by immunohistochemistry. CpG island methylation is a frequent event, occurs early, and accumulates during multi-step prostatic carcinogenesis. High levels of CpG island hypermethylation might serve as a potential biological marker for aggressive prostate cancer. PMID- 14743507 TI - Disrupted traffic of connexin 43 in human testicular seminoma cells: overexpression of Cx43 induces membrane location and cell proliferation decrease. AB - Connexins, the constitutive proteins of gap junctions, are considered to be tumour suppressive agents and are often impaired in the tumourigenic processes. In the present study, the expression of connexin 43 (Cx43), which is involved in the control of spermatogenesis through Sertoli/germ cell coupling, has been investigated in human testicular seminoma cells (tumours and the JKT-1 cell line). Cx43 was immunolocalized in the Golgi apparatus without membrane expression and was detected by immunoblotting in JKT-1 as exclusive 70 kD bands. No mutation could be found by sequencing the transcript obtained by RT-PCR. Transfection with a Cx43-V5 vector reproduced the same gel shift, identifying these 70 kD bands as Cx43. The Cx43-70 kD bands were also expressed in normal testicular tissue, associated with the classical 43 kD isoforms. Stable transfection of JKT-1 with a Cx43-GFP vector allowed restoration of Cx43 membrane expression, functional cell coupling, and inhibition of the cell proliferation rate. Storage of Cx43 in the Golgi apparatus may correspond during spermatogenesis to an intermittent physiological process that becomes permanent in malignant seminoma cells as a result of the tumourigenic process. By preventing Cx43 membrane expression, this disrupted traffic may itself participate in tumour promotion. PMID- 14743508 TI - BRAF mutations are associated with some histological types of papillary thyroid carcinoma. AB - Mutations in the BRAF gene have recently been detected in a wide range of neoplastic lesions with a particularly high prevalence in melanoma and papillary thyroid carcinoma (PTC). The hot-spot mutation BRAF(V599E) is frequently detected in PTC (36-69%), in contrast to its absence in other benign or malignant thyroid lesions. In order to unravel whether there is any association between the occurrence of the BRAF mutation and the histological pattern of PTC, in this study a previous series of 50 PTCs was extended to 134 cases, including ten cases of PTC-related entities-hyalinizing trabecular tumour (HTT) and mucoepidermoid carcinoma (MEC). Using PCR/SSCP and sequencing, the BRAF(V599E) mutation was detected in 45 of the 124 PTCs (36%). No mutations were detected in any case of HTT and MEC. BRAF(V599E) was present in 75% of Warthin-like PTCs and 53% of conventional PTCs, whereas no BRAF(V599E) mutations were detected in any of the 32 cases of the follicular variant of PTC. BRAF(V599E) was also detected in 6 of 11 cases of the oncocytic variant of PTC that displayed a papillary or mixed follicular-papillary growth pattern and in none of the four oncocytic PTCs with a follicular growth pattern. A distinct mutation in BRAF (codon K600E) was detected in three cases of the follicular variant of PTC. This study has confirmed the high prevalence of BRAF(V599E) in PTC and has shown that the mutation is almost exclusively seen in PTC with a papillary or mixed follicular-papillary growth pattern, regardless of the cytological features of the neoplastic cells. The results support the existence of an oncocytic variant of PTC that should be separated from the oncocytic variant of follicular carcinoma and suggest that the follicular variant of PTC may be genetically different from conventional PTC. PMID- 14743509 TI - Loss of heterozygosity at chromosome 9p21 is a frequent finding in enteropathy type T-cell lymphoma. AB - Enteropathy-type T-cell lymphoma (ETL) and ulcerative jejunitis (UJ) are rare disorders often occurring in patients with coeliac disease. The genetic events associated with the accumulation of intraepithelial lymphocytes in coeliac disease and tumour development are largely unknown. Deletions at chromosome 9p21, which harbours the tumour suppressor genes p14/ARF, p15/INK4b, and p16/INK4a, and 17p13, where p53 is located, are associated with the development and progression of lymphomas. To examine whether deletions at 9p21 and 17p13 play a role in ETL, 22 cases of ETL and seven cases of UJ were screened for loss of heterozygosity (LOH) by tissue microdissection and polymerase chain reaction (PCR) analysis for microsatellite markers. Furthermore, p53 and p16 protein expression was examined by immunohistochemistry. In addition, polymerase chain reaction-single strand conformational polymorphism (PCR-SSCP) analysis for detection of mutations in exons 5-8 of the p53 gene was performed in five cases of ETL and three cases of UJ. LOH was found in at least one microsatellite marker at the 9p21 locus in 8 of 22 (36%) ETLs, but not in UJ. Five of nine (56%) tumours composed of large cells showed LOH at 9p21, as opposed to two of eight (25%) tumours with small- or medium-sized cell morphology. The region spanning the p14/p15/p16 gene locus was most frequently affected (five cases); LOH at these markers coincided with loss of p16 protein expression in all of these cases. p53 overexpression was demonstrated in all ETLs examined and in four of seven cases of UJ. However, no alterations of the p53 gene were detected by LOH or PCR-SSCP analysis. The results of this study show that LOH at chromosome 9p21 is frequent in ETL, especially in tumours with large cell morphology; this finding suggests that gene loss at this locus may play a role in the development of ETL. PMID- 14743511 TI - Cyclic AMP-dependent tyrosine phosphorylation in tammar wallaby (Macropus eugenii) spermatozoa. AB - Despite considerable advances in our understanding of the molecular mechanisms regulating eutherian sperm function, there is a paucity of such knowledge for the Metatheria. In eutherian spermatozoa, the attainment of functional competence is associated with a redox-regulated, cAMP-mediated tyrosine phosphorylation cascade, activated during capacitation. In this report we investigate whether tammar wallaby (Macropus eugenii) spermatozoa possess a similar signal transduction pathway. Western blot analysis of phosphotyrosine expression in caudal and ejaculated populations of tammar spermatozoa revealed that elevation of intracellular cAMP levels, but not exposure to oxidants or NADPH, induced a dramatic increase in the overall level of tyrosine phosphorylation. Washed, ejaculated spermatozoa exhibited more pronounced increases in tyrosine phosphorylation than unwashed sperm populations. Localisation of tyrosine phosphorylation by immunocytochemistry showed that phosphotyrosine residues were principally located along the tammar sperm flagellum, and occasionally at a small region of the sperm head, adjacent to the acrosome. Associated with the tyrosine phosphorylation of tammar spermatozoa, was a change in sperm head conformation to a T-shaped orientation, further implying the importance of these pathways to normal tammar sperm function. Redox activity, as detected by lucigenin-dependent chemiluminescence, was stimulated by NADPH in caudal sperm preparations but not ejaculated spermatozoa. However, neither sperm population responded to treatment with NADPH with changes in intracellular cAMP or tyrosine phosphorylation. In conclusion, tammar spermatozoa possess the same cAMP-mediated, tyrosine phosphorylation-dependent signal transduction cascade that has been associated with capacitation in eutherian spermatozoa. However in Metatherian spermatozoa we could find no evidence that this pathway was redox regulated. PMID- 14743512 TI - Pluripotency of cryopreserved blastomeres of the goldfish. AB - To examine the pluripotency of cryopreserved blastomeres, we transplanted them into blastula. Donor blastomeres were prepared from blastula of goldfish (Carassius auratus) and cryopreserved in liquid nitrogen for two months. Fifty five percent and 44% of blastomeres survived after thawing. Cryopreserved blastomeres were transplanted to the blastula of triploid crucian carp (C. a. longsdorfii), which reproduces gynogenetically in nature. At four days after the operation, resultant chimeric embryos transplanted with cryopreserved blastomeres showed a survival rate (41.6%) lower than that of embryos transplanted with unfrozen blastomeres (57.1%). Transplanted blastomeres were histologically identified in various organs derived from all three germ layers. A primordial germ cell differentiated from a cryopreserved blastomere was detected in one of the 32 chimeric fish examined. These results suggest blastomeres that survive after cryopreservation retain their pluripotency and are able to differentiate into both somatic and germ cell lines. PMID- 14743513 TI - Green fluorescent protein gene-transfected peafowl somatic cells participate in the development of chicken embryos. AB - This study was performed to investigate whether the embryonic somatic cells are capable of reconstituting and participating in the embryonic development of chickens to produce chimeras. In order to track the migration behavior of the donor cells, a cell line, originally isolated from an Indian peafowl embryo, was fluorescent-labeled by transfection of the cells with enhanced Green Fluorescent Protein (GFP) and Neomycin resistant (Neo) genes prior to injection into the stage X blastoderm of White Leghorn chickens. The injection was performed with a medium in the presence of 1-5% polyethylene glycol. The development of putative chimeric embryos between the stages three and 24 was examined for GFP expression under fluorescent light. To trace the peafowl cells in the developing chicken embryos, both a species-specific genetic marker originating from the mitochondrial DNA cytochrome b (cyt b) gene and a DNA fragment of GFP gene were used. Of the 185 fertile eggs manipulated, 173 developed into embryos. Fifty-five of them showed positive GFP patches in extra-embryonic tissues, and 15 expressed GFP in intra-embryonic tissues such as those of the head, heart, and gonad. PCR analysis revealed that PCR fragments for the peafowl mitochondrial DNA cyt b and GFP genes were detected in the samples of the GFP positive extra- and intra embryonic tissues of the chimeras. The present results provide evidence that fluorescent-labeled peafowl embryonic cells carrying GFP and Neo genes are able to participate in the development of chicken embryos to generate chimeras. PMID- 14743514 TI - Forelimb spike regeneration in Xenopus laevis: Testing for adaptiveness. AB - Experiments were designed to test adaptability of forelimb spike regenerates in Xenopus laevis froglets. The results show that when amputation is at the radius/ulna level, regeneration occurs in 100% of the cases and a single spike of cartilage is the result. The spike regenerates originating from radius/ulna level amputations can be used for feeding and froglet growth is only minimally compromised by the spike. The spike grows in length as the froglet body grows and thus is in homeostasis with the body. The spike develops nuptial pad tissue in reproductively mature males and is occasionally molted, indicating responsiveness to gonadal and thyroid hormones. Finally, and most important, the spike can be used for amplexus and successful mating. In contrast, spikes originating from humerus level amputations were considerably shorter and regeneration from that limb level was less frequent. When amputation was at the body wall regeneration did not occur. PMID- 14743515 TI - Lymphoid aggregates in gonads of embryos, hatchlings, and young of turtles with temperature-dependent sex determination. AB - Cellular infiltrations forming lymphoid-like aggregates were previously observed in gonads of two turtle species exhibiting temperature-dependent sex determination (TSD): at hatching in Chelydra serpentina; at and after hatching in Emys orbicularis. We show here that such aggregates are also present in gonads of Testudo graeca by the end of embryonic development, suggesting that their occurrence is general in turtles. Since in C. serpentina, infiltrations were observed mainly in testes exhibiting remnants of the germinal epithelium, it was assumed that their occurrence was an expression of maleness leading to rejection of this epithelium. The generality of this hypothesis was tested in E. orbicularis by looking for lymphoid-like aggregates in three types of gonads (testes, ovotestes, and ovaries) and for the stages at which they occur. Gonads were from embryos, hatchlings, and young incubated at various temperatures. Ovotestes obtained by treatment with an aromatase inhibitor of eggs incubated at female-producing temperature were also examined. In these gonads, the differentiation of Sertoli cells in testicular cords/tubes was ascertained by expression of SOX9. Moreover, the cell composition of aggregates was determined on electron micrographs. Aggregates appear in ovaries and ovotestes by the end of embryonic development and are present in the majority of these gonads at hatching, and at least up to one year after hatching. They are composed mainly of lymphocytes and fibroblasts. Aggregates are not present in typical testes. Since they occur in most ovaries, they cannot be seen as an expression of maleness. Rather, lymphocytic infiltration and formation of lymphoid aggregates in turtle gonads can be seen as components of the immune system, and can be under the control of gonadal endogenous sex steroids. PMID- 14743516 TI - Physiology of hibernation in Rana pipiens: metabolic rate, critical oxygen tension, and the effects of hypoxia on several plasma variables. AB - Rates of O(2) consumption (.VO(2)) were determined for adult northern leopard frogs (Rana pipiens) submerged at 3 degrees C at water PO(2)s (P(w)O(2)) ranging from 0-160 mmHg. The critical O(2) tension (P(c)) was 36.4 mmHg. Hematocrit and blood levels of PO(2), glucose, lactate, pH, [Na(+)], [K(+)], and osmolality were determined for frogs submerged for two days. Above a P(w)O(2) of 50 mmHg, blood PO(2) ranged from 1-7 mmHg, which was sufficient to allow the frogs to function entirely aerobically. Plasma [lactate] increased as P(w)O(2) fell below 50 mmHg, the increase preceding significant changes in any other variable, and apparently preceding a fall in .VO(2). Most other variables showed little or no change from those of air-breathing control animals, even during anoxia. We present an analysis of the importance of a large decrease in P(c) in permitting frogs to successfully overwinter in icebound ponds and of the factors that contribute to that decrease. PMID- 14743517 TI - From sneaker to parental male: change of reproductive traits in the black goby, Gobius niger (Teleostei, Gobiidae). AB - This study focuses on the consequences of the switch of tactic from parasitic to parental male in the black goby, Gobius niger (Teleostei: Gobiidae), a species showing two alternative male mating tactics. Older and larger males defend nests, court, and perform parental care on eggs, while younger and smaller ones behave as parasites, sneaking into nests while spawning occurs. Males adopting different tactics are known to present differences in primary and secondary sex traits. The social context of sneaker males was manipulated to induce a tactic switch. Sneakers were kept under two different experimental treatments with or without a female, and under exclusion of male-male competition. Males changed tactics, courting females, spawning, and performing parental care. All males showed substantial changes in primary sexual traits, such as a reduction in gonadal development and an increase in the investment in accessory structures. The experimental groups differed in the functionality of gonads and accessory organs and in the development of the secondary sex traits. These results demonstrate that the moment of switching is not genetically fixed in the black goby. Sneaker males are able to quickly reallocate energy in primary and secondary sex traits, in accordance with the adopted tactic. Several aspects of this flexible reproductive pattern resemble the socially controlled sex change found in sequential hermaphrodites. PMID- 14743518 TI - Daily organization of laying in Japanese and European quail: effect of domestication. AB - In the present study, we aimed to examine the possible effects of domestication on the daily organization of laying in female quail. To achieve this we compared laying patterns in a laboratory strain of Japanese quail with those in European quail originating from a wild population. Under LD 14:10, the same specific laying window was observed, with ovipositions occurring mainly between 7 h and 15 h after lights on. Thus, domestication did not modify the specific daily organization of laying in females laying in the afternoon. Research about the functional value of this temporal organization in the field thus seems justified. At an individual level, two distinct laying profiles were recorded, as in a previous experiment on an industrial Japanese quail strain selected for meat and egg production. However, different frequencies were noted. Whereas stable profile females (laying at the same time each day) were predominant (80.4%, N=102) in the industrial Japanese quail strain, delayed profile females (laying successively later each day) appeared dominant in the European quail (81%, N=22). The Japanese quail from the laboratory strain constituted an intermediate group with a slight dominance of the stable profile (58.8%, N=17). Thus, domestication has changed the frequencies of the laying profiles, favoring stable females, which are the most productive birds. A model involving interaction between one circadian rhythm and one ultradian rhythm can explain the observed laying profiles. PMID- 14743519 TI - Aggregation effects on anhydrobiotic survival in the tardigrade Richtersius coronifer. AB - For anhydrobiotic metazoans the rate of desiccation is an important factor influencing the probability of survival in a dry anhydrobiotic state. Formation of animal aggregations, in which the exposed body surface area of individual animals is reduced, represents one way to reduce the rate of evaporation. Such aggregations have earlier been documented in e.g., nematodes. We experimentally evaluate the effect of aggregation size (number of animals in a group of desiccating animals) on anhydrobiotic survival in the eutardigrade Richtersius coronifer. The experiment shows that aggregation provides a clear improvement on anhydrobiotic survival. The most likely explanation for this is that aggregated animals were exposed to a lower rate of desiccation. Although the empirical evidence of aggregation in tardigrades is scarce, our study suggests that aggregation could potentially be an important survival factor for tardigrades living in environments characterized by periods of rapid desiccation. PMID- 14743520 TI - Sirolimus-eluting coronary stents. PMID- 14743521 TI - Schizophrenia, drug therapy, and monitoring. PMID- 14743522 TI - Intraabdominal pregnancy after hysterectomy. PMID- 14743523 TI - Mobile telephone use among Melbourne drivers: a preventable exposure to injury risk. PMID- 14743524 TI - Developments in shrinkage control of adhesive restoratives. AB - PURPOSE: This article reviews material properties and application techniques important in minimizing effects of polymerization shrinkage during the curing reaction of resin composite restorative materials used in adhesive dentistry. MATERIALS AND METHODS: Relevant scientific publications were critically reviewed. RESULTS: Since it was recognized that shrinkage, which takes place during the curing reaction of resin composite restorative materials, may cause severe problems in adhesive dentistry, considerable effort has been put into reducing the negative effects. The most important problem is the debonding of the restoration-tooth interface, resulting in increased microleakage and, ultimately, in secondary caries. Despite all efforts, there is still no material or general application method that guarantees a leak-proof and durable restoration. CLINICAL SIGNIFICANCE: It is of the utmost importance that dental practitioners know how to deal with the problems related to resin composite shrinkage, so that they can choose the material and procedure most likely to produce a leak-proof and durable restoration, maximizing the potential for clinical success. PMID- 14743525 TI - Curing dental resins and composites by photopolymerization. AB - The development and continued evolution of photopolymerizable dental materials, particularly dental composite restoratives, represent a significant, practical advance for dentistry. The highly successful integration of the light-activated curing process for dental applications is described in this review. The basic mechanisms by which the photoinitiators efficiently convert monomers into polymers are discussed along with the variety of factors that influence the photopolymerization process. The conventional camphorquinone-amine visible light photoinitiator system used in most dental restorative materials is illustrated in addition to some alternative initiator systems that have been studied for dental materials applications. CLINICAL SIGNIFICANCE: Photopolymerization has become an integral component of the practice of dentistry. A better appreciation of the photopolymerization process as well as its potential and limitations may aid the dentist in the delivery of both esthetic and restorative dental care. PMID- 14743526 TI - Polymerization contraction stress of resin composite restorations in a model Class I cavity configuration using photoelastic analysis. AB - PURPOSE: An important factor that contributes to deterioration of resin composite restorations is contraction stress that occurs during polymerization. The purpose of this article is to familiarize the clinician with the characteristics of contraction stress by visualizing the stresses associated with this invisible and complex phenomenon. MATERIALS AND METHODS: Internal residual stresses generated during polymerization of resin composite restorations were determined using micro photoelastic analysis. Butt-joint preparations simulating Class I restorations (2.0 mm x 5.0 mm, 2.0 mm in depth) were prepared in three types of substrates (bovine teeth, posterior composite resin, and transparent composite resin) and were used to examine contraction stress in and around the preparations. Three types of composite materials (a posterior composite, a self-cured transparent composite, and a light-cured transparent composite) were used as the restorative materials. The self-cured composite is an experimental material, and the others are commercial products. After treatment of the preparation walls with a bonding system, the preparations were bulk-filled with composite. Specimens for photoelastic analysis were prepared by cutting sections perpendicular to the long axis of the preparation. Fringe patterns for directions and magnitudes of stresses were obtained using transmitted and reflected polarized light with polarizing microscopes. Then, the photoelastic analysis was performed to examine stresses in and around the preparations. RESULTS: When cavity preparations in bovine teeth were filled with light-cured composite, a gap was formed between the dentinal wall and the composite restorative material, resulting in very low stress within the restoration. When cavity preparations in the posterior composite models were filled with either self-cured or light-cured composite, the stress distribution in the two composites was similar, but the magnitude of the stress was greater in the light-cured material. When preparations in the transparent composite models were filled with posterior composite and light-cured transparent composite material, significant stress was generated in the preparation models simulating tooth structure, owing to the contraction of both restorative materials. CLINICAL SIGNIFICANCE: Polymerization contraction stress is an undesirable and inevitable characteristic of adhesive restorations encountered in clinical dentistry that may compromise restoration success. Clinicians must understand the concept of polymerization contraction stress and realize that the quality of composite resin restorations depends on successful management of these stresses. PMID- 14743527 TI - Effect of distance on the power density from two light guides. AB - PURPOSE: This study determined the effect of distance on the power density from standard and Turbo light guides (Demetron/Kerr, Danbury, Connecticut). MATERIALS AND METHODS: Power density was measured from 0 to 10 mm away from the tip of standard 8-mm curved light guides and 13/8-mm Turbo curved light guides. To determine the effect of distance on power density, a polynomial regression line was fitted. The Kolmogorov-Smirnov (K-S) statistic and the Wilcoxon rank sum (WR) tests were used to determine if there was a difference in the rate at which the power density decreased for the standard and Turbo light guides as the distance from the tip increased. Photographs of the light dispersion from each tip were also taken. RESULTS: At 0 mm, the mean (+/- SD) power density from the two standard light guides was 743 +/- 6.1 mW/cm2 and from the four Turbo light guides was 1128 +/- 22.1 mW/cm2. As the distance from the tip of the light-guide tip increased, the power density decreased, but the rate of decrease was greater from the Turbo light guides than from the standard light guides. At 6 mm the power density from the standard light guides fell to 372 mW/cm2 (50% of the original value) and the power density from the Turbo light guides fell to 263 mW/cm2 (23% of the original value). Both the K-S statistic and the WR sum test indicated that the distribution of light intensities was significantly different from the two light guides (WR p-value = .0246, K-S p-value < .0001). The two estimated polynomials intersected at 3.66 mm, and the 95% prediction intervals intersected at about 2.8 and 4.8 mm. Therefore, beyond 5 mm away from the tip of the light guide, the standard light guides gave higher power density readings than the Turbo light guides. Photographs showed that the light dispersed at a wider angle from the Turbo light guides than from the standard light guide. CLINICAL SIGNIFICANCE: The design of the light guide of a light curing unit affects light dispersion, power density, and ultimately the dentist's ability to properly cure composite. For these reasons, manufacturers should report the power density at the tip of the light guide and 6 mm from the tip of the light guide, since significant differences exist between light guide designs. PMID- 14743528 TI - Effect of ramped light intensity on polymerization force and conversion in a photoactivated composite. AB - PURPOSE: This study evaluated the effect of ramped light intensity on the polymerization shrinkage forces and degrees of conversion (DC) of a hybrid composite. MATERIALS AND METHODS: Composite samples were bonded between two steel rods (2.50 mm diameter, 1.25 mm apart, configuration factor = 1.0) mounted in a universal testing machine using a constant displacement mode. Polymerization contraction force was recorded for 250 seconds under four light exposure conditions: group 1, STD: (40 s x 800 mW/cm2); group 2, EXP: (150 mW/cm2 logarithmic increase to 800 mW/cm2 over 15 s) + (25 s x 800 mW/cm2); group 3, 2 STEP: (10 s x 150 mW/cm2) + (30 s x 800 mW/cm2); group 4, MED: (80 s x 400 mW/cm2). Maximum curing force (N250s) and maximum force rate of the four groups were compared using one-way analysis of variance (ANOVA) (alpha = 0.05) and the Tukey test. Degrees of conversion obtained with STD, EXP, and MED cure modes were evaluated at three depths (top surface, 1 mm, and 2 mm) using Fourier transform infrared spectroscopy (FTIR). RESULTS: Maximum rates of polymerization shrinkage force development and standard deviations (SD), in ascending order, were group 4, MED: 0.33 +/- 0.03 N/s; group 2, EXP: 0.35 +/- 0.06 N/s; group 1, STD: 0.44 +/- 0.03 N/s; and group 3, 2-STEP: 0.46 +/- 0.07 N/s. Maximum rates of polymerization shrinkage force development of group 2, EXP and group 4, MED were statistically equivalent and lower than those of group 1, STD and group 3, 2-STEP. Maximum shrinkage forces (+/- SD), in ascending order, were group 2, EXP: 20.4 +/- 2.5 N; group 4, MED: 25.8 +/- 1.0 N; group 3, 2-STEP: 27.4 +/- 5.8 N, and group 1, STD: 30.5 +/- 2.7 N. Maximum force of the EXP mode was statistically lower than MED, 2 STEP, and STD curing modes. The EXP ramp was successful in reducing the conversion rate at the top surface and at 1.0-mm depth, but it did not affect the total conversion compared to the STD 40-second cure mode. There was no difference in DC at the top surface and 1-mm depth with mode of cure. The MED cure mode resulted in a higher DC than the EXP mode at a depth of 2 mm. CLINICAL SIGNIFICANCE: Maximum shrinkage force and force rate exhibited during the first 250 seconds of polymerization were significantly lower using a ramped light intensity exposure. Ramped light intensity decreased conversion rate at the top surface and at 1.0-mm depth and did not affect the total extent of conversion compared to a standard 40-second, single-intensity cure mode. The slower conversion rate resulting from ramped light intensity helped to reduce the rate and maximum polymerization stress, but would not be expected to compromise the physical properties for the restorative material, since similar degrees of conversion were obtained. PMID- 14743529 TI - Polymerization depths of contemporary light-curing units using microhardness. AB - PURPOSE: This research investigated composite depths of cure using a variety of light-curing units and exposure protocols. MATERIALS AND METHODS: Composite (Herculite XRV, shade A2, Kerr, Orange, California) was exposed in opaque compules to conventional quartz tungsten halogen (QTH) units, soft-start units, high-intensity QTH and plasma arc (PAC) curing lights, and one argon laser. Cured compules were sonicated to remove uncured composite and were sectioned and polished along the long axis to expose cured composite. Knoop hardness was measured 0.5 mm from the irradiated, top surface and then at 1.0 mm and in 1.0-mm increments until reliable readings could no longer be obtained. Hardness values were compared by analysis of variance at similar depths within a specific curing light classification, using the hardness of the standard 40-second conventional QTH exposure as comparison (Dunnett's t-test). Depth of cure was defined as the deepest hardness value found equivalent to that at 0.5-mm depth for a specific curing light and scenario. RESULTS: Conventional QTH lights provided similar hardness profiles. At 2-mm depth, use of a different unit or curing tip made no difference in hardness compared with the standard. At this depth, soft-start (pulse-delay and step-cure) methods yielded hardness similar to that of the standard. High-intensity QTH lights provided similar hardness at 2-mm depth in 10 seconds to that of the standard 40-second exposure. Plasma arc exposure for less than 10 seconds produced inferior hardness compared with the standard. A 10 second PAC and a 5-second laser exposure gave hardness at 2-mm depth equivalent to that of the 40-second standard. Depth of cure for almost all curing scenarios was not greater than 2 mm. CLINICAL SIGNIFICANCE: Similar-type conventional QTH lights with different tip diameter (8 and 12 mm) provide similar composite cure characteristics. Soft-start techniques provide similar cure profiles to those achieved with conventional QTH technique when used according to manufacturer's recommendations. High-intensity QTH units and the argon laser can reduce exposure time while providing composite with similar hardness to that of conventional QTH curing. Plasma arc exposure should be at least of 10 seconds duration to provide hardness equivalent to that achieved with conventional 40-second QTH exposure. Even with consideration of high-intensity curing units, composite increments should still be no greater than 2 mm to provide homogeneous hardness. PMID- 14743530 TI - [Glutamyl endopeptidase. Structure, function, practical use]. AB - Special features of the structural organization of serine proteases belonging to a new subfamily of glutamyl-specific endopeptidases, which possess an extremely strict substrate specificity, are discussed. Some areas of the practical application of these enzymes are considered. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2003, vol. 29, no. 6; see also http://www.maik.ru. PMID- 14743531 TI - [Interaction of cardiotoxin A5 with a membrane: role of conformational heterogeneity and hydrophilic properties]. AB - The hypothesis that local conformational differences of snake venom cardiotoxins (cytotoxins, CTs) may play a significant role in their interaction with membrane was tested by molecular modeling of the behavior of the CT A5 from the venom of Naja atra in water and at the water-membrane interface. Two models of the CT A5 spatial structure are known: the first was obtained by X-ray analysis and the second, by NMR studies in solution. A molecular dynamics (MD) analysis demonstrated that loop II of the toxin has a fixed omega-like shape in water, which does not depend on its initial structure. Interaction of the experimentally derived (X-ray and NMR) conformations and MD-simulated conformations of CT A5 with the lipid bilayer was studied by the Monte Carlo method using the previously developed model of the implicit membrane. The following was found: (1) Unlike the previously studied CT2 from the venom of cobra Naja oxiana, CT A5 has only loops I and II bound to the membrane, with the involvement of a lesser number of hydrophobic residues. (2) A long hydrophobic area is formed on the surface of CT A5 due to the omega-like shape of loop II and the arrangement of loop I in proximity to loop II. This hydrophobic area favors the toxin embedding into the lipid bilayer. (3) The toxin retains its conformation upon interaction with the membrane. (4). The CT A5 molecule has close values of the potential energy in the membrane and in an aqueous environment, which suggests a dynamic character of the binding. The results of the molecular modeling indicate a definite configuration of loops I and II and, consequently, a specific character of distribution of polar and apolar properties on the toxin surface, which turns out to be the most energetically favorable. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2003, vol. 29, no. 6; see also http://www.maik.ru. PMID- 14743532 TI - [Epitope of the catalytic subunit of viscumin, exposed on its interaction with a lipid bilayer]. AB - It was previously shown that the catalytic subunit of the plant toxin viscumin induces aggregation of small unilamellar liposomes and this process is inhibited by the mab_TA7 monoclonal antibody produced to the denatured catalytic subunit of viscumin (Agapov, I.I. et al., FEBS Lett., 1999, vol. 464, pp. 63-66). The interaction of the synthetic F101-T105 and A96-T105 fragments of the viscumin catalytic subunit with the mab_TA7 monoclonal antibody was studied by 1H NMR spectroscopy. The results of this study demonstrated that only the A96-T105 fragment is capable of binding to mab_TA7. A nuclear Overhauser effect observed in the antigen-antibody complex and registered on the resonances of the free peptide and exchanging between the free state and the antibody-bound state was analyzed; the mab_TA7 antigen determinant (H99-T105) was identified; and its conformation and orientation within the complex with the antibody were determined. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2003, vol. 29, no. 6; see also http://www.maik.ru. PMID- 14743533 TI - [Catechol siderophore, produced by thermoresistent strain of Bacillus licheniformis VK21]. AB - Thermophilic and thermoresistant strains of bacilli were screened on a medium containing Chrome Azurol S for producers of siderophores. It was found that the Bacillus licheniformis VK21 strain dramatically increases secretion of the metabolite, a chelator of Fe3+, in response to addition of manganese(II) salts. The growth of the producer on a minimum medium containing MnSO4 under the conditions of iron deficiency is accompanied by the accumulation of a catechol product, the content of which reaches a maximum at the beginning of the stationary growth phase of culture. In the presence of FeCl3, the amount of the catechol product in the medium considerably decreases. The siderophore, called SVK21, was isolated from the cultural medium and purified by reversed phase HPLC, and its siderophore function was confirmed by the test for the restoration of growth of producer cells in a medium containing EDTA. The UV spectrum of the siderophore has absorption maxima at 248 and 315 nm. According to amino acid analysis and NMR spectrometry, the metabolite SVK21 is 2,3-dihydroxybenzoyl glycyl-threonine. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2003, vol. 29, no. 6; see also http://www.maik.ru. PMID- 14743534 TI - [Interaction of duodenase with human blood serpins]. AB - The interaction of duodenase, a new serine protease from a small group of Janus faced proteases, with serpins, alpha 1-protease inhibitor (alpha 1-PI) and antichymotrypsin (ACT) from human blood serum, was studied. The stoichiometry of the inhibition process was found to be 1.2 and 1.3 mol/mol for alpha 1-PI and ACT, respectively. The presence of a stable enzyme-inhibitory complex duodenase alpha 1-PI was confirmed by SDS-PAGE. No formation of the duodenase-ACT complex was demonstrated; instead, the band of the cleaved inhibitor was indicated upon the ACT hydrolysis. The suicide mechanism of the duodenase interaction with the human blood serpins was proved. The association rate constants (Ka, M-1 s-1) were 2.4 +/- 0.3 x 10(5) for alpha 1-PI and 3.0 +/- 0.4 x 10(5) for ACT. These results indicate the possibility of the regulation of duodenase activity by endogenous serpins. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2003, vol. 29, no. 6; see also http://www.maik.ru. PMID- 14743535 TI - [Hydrolysis of phosphoether bonds of heme-independent chloride peroxidase from Serratia marcescens]. AB - Heme- and metal-independent chloroperoxidase from Serratia marcescens W 250 is shown to be capable of catalyzing the p-nitrophenyl phosphate hydrolysis. The parameters of the phosphatase reaction are determined and inhibitors and activators of the process are found. A hypothetical mechanism of the hydrolysis of phosphoesters by heme- and metal-independent haloperoxidases is suggested. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2003, vol. 29, no. 6; see also http://www.maik.ru. PMID- 14743536 TI - [Chemical and chemico-enzymatic synthesis of alpha-thiotriphosphate nucleosides]. AB - New methods of chemical and chemoenzymatic synthesis of nucleoside 5' thiophosphates and 5'-alpha-thiotriphosphates are developed. The 5'-alpha thiotriphosphates are used as substrates both in template-dependent enzymatic PCR synthesis and in a T7-RNA transcription polymerase system. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2003, vol. 29, no. 6; see also http://www.maik.ru. PMID- 14743537 TI - [Preparation of DNA-duplexes, containing internucleotide thiophosphate groups in various positions of the recognition site for the EcoRII restriction endonuclease]. AB - Twenty-four 12-mer DNA duplexes, each containing a chiral phosphorothioate group successively replacing one of the internucleotide phosphate groups either in the EcoRII recognition site (5'CCA/TGG) or near to it, were obtained for studying the interaction of the restriction endonuclease EcoRII with internucleotide DNA phosphates. Twelve of the 12-mer oligonucleotides were synthesized as Rp and Sp diastereomeric mixtures. Six of them were separated by reversed-phase HPLC using various buffers. Homogeneous diastereomers of the other oligonucleotides were obtained by enzymatic ligation of the Rp and Sp diastereomers of 5- to 7-mer oligonucleotides preliminarily separated by HPLC with the corresponding short oligonucleotides on a complementary DNA template. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2003, vol. 29, no. 6; see also http://www.maik.ru. PMID- 14743538 TI - [Copper-catalyzed cleavage of DNA by arenes]. AB - DNA was found to be cleaved in neutral solutions containing arenes and copper (II) salts. The reaction is comparable in efficiency with the DNA cleavage by such systems as Cu(II)-phenanthroline and Cu(II)-ascorbic acid, but, in contrast to the latter, the system Cu(2+)-arene does not require the presence of an exogenous reducing agent or hydrogen peroxide. The system Cu(2+)-arene does not cleave DNA under anaerobic conditions. Catalase, sodium azide, and bathocuproine, which is a specific chelator of Cu(I), completely inhibit the reaction. The data obtained allow one to suppose that Cu(I) ions, superoxide radical, and singlet oxygen participate in the reaction. It has been shown by the EPR method using spin traps that the reaction proceeds with formation of alkoxyl radicals, which can insert breaks in the DNA molecule. For effective cleavage of DNA in the Cu(II)-o-bromobenzoic acid system, the radicals have to be generated by a specific copper-DNA-o-bromobenzoic acid complex, in which copper ions are most probably coordinated with oxygen atoms of the DNA phosphate groups. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2003, vol. 29, no. 6; see also http://www.maik.ru. PMID- 14743539 TI - [Oxidation of 17alpha,20beta- and 17alpha,20alpha-dihydroxysipregn-4-en-ones- side products of biotransformation of progesterone by recombinant microorganisms expressing cytochrome P45017alpha]. AB - Progesterone biotransformation with recombinant yeast Yarrowia lipolytica E129A15 and Saccharomyces cerevisiae GRF18/YEp5117 alpha expressing bovine adrenocortical cytochrome P45017 alpha yielded 17 alpha-hydroxyprogesterone and two diols, 17 alpha, 20 beta- and 17 alpha, 20 alpha-dihydroxypregn-4-en-3-one. The oxidation of mixtures of the three steroids with chromic acid resulted in the cleavage of 17-20 bonds in the diols with the formation of androst-4-ene-3,17-dione. The biotransformation of pregn-4-ene-20 beta-ol-3-one by means of Y. lipolytica E129A15 was accompanied by the following reactions: the primary oxidation of these compounds to progesterone and the subsequent successive reactions of 17 alpha-hydroxylation and 20 alpha- and 20 beta-reduction. The results widen the possibilities for enzymatic and chemical modifications of steroids. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2003, vol. 29, no. 6; see also http://www.maik.ru. PMID- 14743540 TI - [Transformation of 3beta-(2-hydroxyethoxy)-5alpha-cholest-8(14)-en-15-one in a polar metabolite in HepG2 hepatoma cells]. AB - Incubation of 3 beta-(2-hydroxy-2[3H]-ethoxy)-5 alpha-cholest-8(14)-en-15-one with Hep G2 cells led to the accumulation of a radioactive polar product in the culture medium, which was identified as 3 beta-(2-hydroxyethoxy)-15-keto-5 alpha cholest-8(14)-ene-24-oic acid. Its structure was confirmed by a chemical counter synthesis. The labeled ketosterol was rapidly (tau 1/2 = 6 min) and reversibly bound by Hep G2 cells. The intracellular concentration of 15-ketosterol decreased during incubation mainly due to the formation of a polar metabolite, secreted to the medium. The level of cholesterol biosynthesis was 22 +/- 5% of the control value in Hep G2 cells at a 15-ketocholesterol concentration in the medium of 30 microM. However, further incubation for 3 h in the medium without the ketosterol led to restoration of the level of biosynthesis to 85 +/- 11% of the control value. These results suggest that inhibition of the cholesterol biosynthesis by 15-ketocholesterol in Hep G2 cells depends on the intracellular concentration of the inhibitor, which, in turn, is determined by the rate of its conversion into the polar metabolite. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2003, vol. 29, no. 6; see also http://www.maik.ru. PMID- 14743541 TI - [Synthesis and antiviral activity of ureides and carbamates of betulinic acid and its derivatives]. AB - Ureides and carbamates of betulinic acid and its derivatives were prepared in good yields by interaction of betulinic acid, betulonic acid, and betulonic acid 3-oxime with amines, amino acids, and alcohols. Ureides of betulonic acid containing L-Val and L-Met residues were found to be effective against herpes simplex type 1 virus. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2003, vol. 29, no. 6; see also http://www.maik.ru. PMID- 14743542 TI - [Transformation of glycyrrhizinic acid. XV. Synthesis of triterpene saponins with monosaccharide residues, attached through complex ester bonds]. AB - Triterpene saponins, glycoside analogues of glycyrrhizic acid with a modified carbohydrate chain containing monosaccharide residues attached through ester bonds, were synthesized. To this end, peracetylated glycyrrhizic acid or its 30 methyl ester were glycosylated by 2,3,4,6-tetra-O-acetyl-alpha-D-gluco-or-alpha-D galactopyranosyl bromide in dichloroethane in the presence of Ag2CO3. Glycerrhetinic acid saponin with D-Galp residues exhibited a higher antiulcer activity than glycyrrhizic acid in rats at a dose of 25 mg/kg. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2003, vol. 29, no. 6; see also http://www.maik.ru. PMID- 14743543 TI - Citrus pectin and oligofructose improve folate status and lower serum total homocysteine in rats. AB - Low folate status leads to increased total homocysteine (tHcy) concentration, and this has been associated with an increased risk of several diseases. Many colonic bacteria are capable of synthesizing folate, and certain dietary fibers may enhance this effect. We assessed the ability of non-fermentable (cellulose) and fermentable (citrus pectin and oligofructose) fibers to improve folate status and lower tHcy in rats. Weanling Sprague-Dawley rats were fed a folate-deficient diet with 5% cellulose for four weeks. Rats were then randomly assigned to one of five folate-adequate (400 micrograms/kg diet) test diets for 24 days. Diets were as follows: Basal; Basal + Sulfa Drug (succinylsulfathiazole); Cellulose; Citrus Pectin; and Oligofructose. High-fiber diets were formulated by diluting the basal diet such that the final diets contained 10% of the added fiber. Twenty-one days later, 3H-p-aminobenzoic acid was injected into the cecum, and rats were terminated three days later. Rats receiving the Citrus Pectin diet had significantly higher plasma (p = 0.011), erythrocyte (p = 0.035), and colonic tissue folate concentrations (p = 0.013) and lower tHcy (p = 0.003) than rats given the Cellulose diet. Rats receiving the Oligofructose had significantly higher plasma folate (p < 0.001) and lower tHcy (p = 0.032) concentrations than rats receiving the Cellulose diet. 3H-folate was detected in the livers of all rats except those receiving Sulfa Drug. Our study indicates that Citrus Pectin and Oligofructose, but not Cellulose, can significantly increase indices of folate status in rats and lower tHcy. It also confirms the ability of the large bowel to absorb folate. PMID- 14743544 TI - Pilot clinical trial of the use of alpha-tocopherol for the prevention of hepatocellular carcinoma in patients with liver cirrhosis. AB - Patients with chronic hepatitis C virus (HCV) infection often develop liver cirrhosis and hepatocellular carcinoma (HCC). The purpose of this study was to test the chemopreventive effect of alpha-tocopherol on hepatocarcinogenesis in patients with liver cirrhosis and a history of HCV infection. Eighty-three patients with liver cirrhosis and with positive history of HCV infection were divided at random into two groups. Forty-four patients were treated with alpha tocopherol (Vit E group) while the other 39 were followed as controls. The clinical background (gender, age, and laboratory data) was similar in the two groups. Serum levels of alpha-tocopherol, albumin, alanine aminotransferase (ALT), and total cholesterol and platelet count were measured serially over a period of five years. The mean serum concentration of alpha-tocopherol was low in both groups at entry and was significantly higher in the Vit E group than in the control group one month after treatment. Platelet count, serum albumin, ALT, and total cholesterol were not different between the two groups during the five-year period. Cumulative tumor-free survival and cumulative survival rate tended to be higher in the Vit E group than in controls, albeit statistically insignificant. The serum level of alpha-tocopherol was low in patients with liver cirrhosis and positive for HCV. Although the administration of alpha-tocopherol normalized the level one month later, it could neither improve liver function, suppress hepatocarcinogenesis, nor improve cumulative survival. Patients treated with alpha-tocopherol tended to live longer without development of HCC but the difference was not statistically significant. PMID- 14743545 TI - Oral administration of ascorbic acid attenuates endothelial dysfunction after short-term cigarette smoking. AB - Short-term cigarette smoking is associated with persistent endothelial dysfunction. Data on the reversibility of this effect with per os antioxidants after short-term smoking are lacking. This study examines the effect of orally administered ascorbic acid on cigarette smoking-induced endothelial dysfunction. In the present double-blind, randomized, crossover study, 19 healthy subjects (28.7 +/- 6.8 years, mean +/- SD) were examined by high-resolution ultrasonography of the brachial artery before and 0, 30, 60, 90, and 120 minutes after smoking a cigarette. Flow-mediated dilatation (FMD) was used as a method to examine endothelial function. Measurements were performed on two different days, 2 hours after oral administration of 2 g of ascorbic acid or placebo. FMD was similar for each subject between the two visits at baseline. FMD was significantly decreased after smoking with both placebo and ascorbic acid. However, there was a significant beneficial effect of ascorbic acid on the FMD change over time after smoking. After smoking, the FMD dropped to less than half of the baseline value. Thereafter in the placebo group, FMD increased to 70% of baseline value in 90 minutes, but in the ascorbic acid group the FMD increased to 70% of baseline value in 46 minutes. Oral administration of ascorbic acid attenuates endothelial dysfunction after short-term cigarette smoking by shortening its duration. PMID- 14743546 TI - Minimal inflammation, acute phase response and avoidance of misclassification of vitamin A and iron status in infants--importance of a high-sensitivity C-reactive protein (CRP) assay. AB - The acute phase response is known to affect many biological parameters used to assess the iron and vitamin A status. Usually, C-reactive protein (CRP) values higher than 5 to 10 mg/L are taken as indicative of this response. Here we report changes occurring at much lower CRP values. A range of parameters (clinical chemistry, retinol, vitamin E, carotenoids, thyroid status, blood count, immunology) were measured in 101 healthy one-year-old children with no or only minimal symptoms of airway infection and CRP values below 6 mg/L on routine testing. Additionally, CRP values were measured by a high-sensitivity assay (detection limit 0.2 mg/L). When determined by a more sensitive assay, CRP values (median, 0.26 mg/L) revealed highly significant associations with parameters known to be influenced by the acute phase response. Using a limit of 0.6 mg/L (75th percentile), significantly lower levels were observed for transthyretin, iron, retinol, and beta-carotene in the group with the higher CRP levels. This relationship was confirmed intra-individually in a subgroup of 21 children who underwent a second blood draw about four weeks after the first one. The acute phase response is triggered at very low inflammatory levels with CRP values considerably lower than 5 to 10 mg/L, and can occur in apparently healthy children. A high-sensitivity CRP assay is necessary to detect this low level, and in the case of iron or vitamin A, it can help to avoid misclassification of the nutritional status. PMID- 14743547 TI - Hyperhomocysteinemia and oxidative stress in hemodialysis: effects of supplementation with folic acid. AB - This study was undertaken to evaluate two different doses of folic acid and their effects on the control of hyperhomocysteinemia, and on pro-oxidant and antioxidant changes in a group of 32 hemodialysis (HD) patients. Blood samples were collected in a group of patients at three different times: before (basal; B), after the first (S1), and after the second (S2) three-month supplementation periods, and compared to samples from a group of healthy individuals. Analysis of vitamins (C, E, folate, and B12), oxidant parameters (lipid and protein oxidation), and homocysteine were performed. Hyperhomocysteinemia of different degrees was observed in all patients on HD (45.30 +/- 24.89 microM). Oxidative stress was also detected, with lipoperoxidation and protein oxidation being associated with lower concentrations of antioxidant substances (vitamins E and C). The first folate dose (2.5 mg after each dialysis session) reduced by half the initial concentrations of homocysteine (44.92 +/- 22.05 to 20.56 +/- 6.79 microM; p < 0.05) but did not normalize its values. The second dose (15 mg) did not show an additional effect, but it was at this time that lipoperoxidation was significantly reduced, although the protein oxidation showed no change. It was concluded that the first dose of folic acid was efficient in reducing homocysteine concentrations, without normalization of values. The participation of hyperhomocysteinemia in oxidative stress appeared to be partial, but in combination with dialysis treatment, may contribute to the induction of an oxidative environment in this group. The possible antioxidant action of folate must also be considered in this case, acting directly against lipoperoxidation or through hyperhomocysteinemia control. Routine supplementations of folic acid and other antioxidant vitamins should be considered in hemodialysis in order to reduce homocysteine levels to lower values, that although not normal, may be more beneficial in minimizing the cardiovascular risk in this group. PMID- 14743548 TI - Influence of maternal education on food consumption and energy and nutrient intake in a group of pre-school children from Madrid. AB - A study was conducted on the influence of maternal education level on food consumption, energy and nutrient intake, and dietary adequacy in 110 pre-school children from Madrid, Spain. With increasing maternal education, children consumed more sugar(p < 0.05), fruit (p < 0.05), and fish (p < 0.05). Snacking was more frequent with decreasing maternal education (p < 0.05). Though statistical significance was not reached, the consumption of pre-cooked foods was greater among children of mothers educated to a higher level, a phenomenon probably related to the work situation of these women. With respect to dietary composition, no significant differences were found between groups for macronutrient, fiber and energy intakes, except for energy supplied by polyunsaturated fatty acids (PUFA), which was greater in the children of less educated women (p < 0.01). This is probably due to their greater consumption of sunflower seed oil. The diets of children belonging to well-educated mothers came closer to meeting the recommended intakes for folate, vitamin C, and iodine. It would seem that maternal educational level influences the food habits of children. Mothers with less education may require special advice in this area. PMID- 14743549 TI - Influence of maternal protein intake on nitrogen fractions of human milk. AB - OBJECTIVES: Studies relating maternal diet to milk composition in humans have led to contradictory conclusions with regard to protein intake. The purpose of this paper was to examine if differences of protein intake in a group of lactating women can impact the protein and non-protein nitrogen fraction of their milk. SUBJECTS AND METHODS: Healthy lactating women(117) were recruited in Northern, Central, and Southern Italy. Their anthropometric measurements were obtained one month after delivery, while dietary evaluation was based on two consecutive 24 hour recalls. Infants' growth parameters were measured at birth and at one month. Milk collected at one month was analyzed for content of total nitrogen, protein nitrogen, non-protein nitrogen, and free amino acid profile. RESULTS: Maternal energy intakes were below the recommended values, while protein intakes were higher, with significant differences between geographical areas. There were no differences in the nitrogenous components of the milk examined with the exception of the non-protein nitrogen fraction and serine. Despite the different maternal intakes, no correlations were found between the mothers' parameters and milk components. CONCLUSION: This study shows that when protein requirements are met and there are no remarkable differences between intakes and requirements, there are no repercussions in nitrogen fractions. PMID- 14743550 TI - Green tea extract suppresses the age-related increase in collagen crosslinking and fluorescent products in C57BL/6 mice. AB - Collagen crosslinking during aging in part results from Maillard reaction endproducts of glucose and oxoaldehydes. Because of the tight link between oxidative and carbonyl stress, we hypothesized that natural antioxidants and "nutriceuticals" could block collagen aging in C57BL/6 mice. Six groups of young and adult mice received vitamin C, vitamin E, vitamin C&E, blueberry, green tea extract (GTE), or no treatment for a period of 14 weeks. Body weights and collagen glycation were unaltered by the treatment. However, GTE or vitamin C&E combined blocked tendon crosslinking at 10 months of age (p < 0.05, adult group). GTE also blocked fluorescent products at 385 and 440 nm (p = 0.052 and < 0.05, respectively) and tended to decrease skin pentosidine levels. These results suggest that green tea is able to delay collagen aging by an antioxidant mechanism that is in part duplicated by the combination of vitamin C and E. PMID- 14743551 TI - Expression of oncogenes depends on biotin in human small cell lung cancer cells NCI-H69. AB - Oncogenes play important roles in cell proliferation and biotin status correlates with gene expression and proliferation rates in human cells. In this study we determined whether biotin supply affects biotin homeostasis, expression of oncogenes, and proliferation rates in NCI-H69 small cell lung cancer cells. NCI H69 cells were cultured in media containing deficient (0.025 nmol/L), physiologic (0.25 nmol/L), or pharmacologic (10 nmol/L) concentrations of biotin for 3 weeks. Biotin concentrations in culture media correlated negatively with biotin transport rates, suggesting that cells responded to marginal biotin supply with increased expression of biotin transporters. Increased biotin uptake was not sufficient to prevent depletion of intracellular biotin in cells cultured in biotin-deficient medium, as judged by decreased activity of biotin-dependent propionyl-CoA carboxylase and decreased biotinylation of histones. The expression of oncogenes N-myc, c-myb, N-ras, and raf correlated with biotin supply in media: oncogene expression increased by up to 20% in cells cultured in pharmacologic medium compared to physiologic controls; oncogene expression decreased by up to 47% in cells cultured in deficient medium. This observation is consistent with a role for biotin in oncogene-dependent metabolic pathways. Cellular uptake of thymidine (marker for proliferation) was not affected by biotin supply, suggesting that effects of biotin-dependent expression of oncogenes on the growth of tumor cells are quantitatively minor. The clinical significance of effects of biotin supply on expression of oncogenes remains to be elaborated. PMID- 14743552 TI - The effect of graded levels of dietary casein, with or without methionine supplementation, on glutathione concentration in unstressed and endotoxin-treated rats. AB - Glutathione (GSH) concentration was measured in rats fed either graded levels of dietary casein (experiment 1; 180 g, 120 g, 80 g, or 60 g protein/kg diet) or graded levels of dietary casein, supplemented with methionine to equalize dietary sulfur amino acid content to that seen in an 180 g/kg casein diet supplemented with 0.3 g L-methionine/kg diet (experiment 2; 180 g protein +0.3 g L-methionine, 80 g protein +6.70 g L-methionine, or 60 g protein +7.45 g L-methionine/kg diet). Rats were given an inflammatory challenge by intraperitoneal injection of endotoxin (lipopolysaccharide from Escherichia coli), and were compared with ad libitum and pair-fed controls. Glutathione concentration in various organs (liver, lung, spleen, and thymus) decreased in animals fed the low-protein diets (80 g or 60 g/kg diet). Addition of the sulfur amino acid, methionine, to the low protein diets restored glutathione concentrations in animals fed ad libitum and prevented the fall in GSH concentration, which occurred in lung, spleen, and thymus in response to the endotoxin. Despite the similarity in the amount of sulfur amino acid consumed between the groups fed the 180 g protein +0.3 g L methionine and the 60 g protein +7.45 g L-methionine/kg diet, in experiment 2, hepatic GSH concentration significantly increased in the latter group, in animals fed ad libitum and in the endotoxin-treated animals, but not in the pair-fed controls. PMID- 14743553 TI - Do we need a Belgian Surgical Journal? PMID- 14743554 TI - Acta Chirurgica Belgica: a challenging task worth the effects? PMID- 14743555 TI - Where should endovascular procedures be performed? PMID- 14743556 TI - The thrombolytic management of bypass graft occlusion. PMID- 14743557 TI - Mechanical thrombectomy in acute and subacute leg ischemia. PMID- 14743558 TI - Endovascular treatment of ruptured infrarenal aortic and iliac aneurysms. PMID- 14743559 TI - Open surgery after endovascular AAA repair. PMID- 14743560 TI - Sleeve lobectomy for non-small cell lung cancer. AB - Sleeve lobectomy is a procedure in which the involved lobe with part of the main stembronchus is removed. The remaining lobe (s) is reimplanted on the main stembronchus. This procedure is indicated for central tumors of the lung as an alternative to pneumonectomy. It is the aim of this study to describe the technique of sleeve lobectomy and to analyse the early postoperative results and late results (survival-recurrence) after sleeve lobectomy for non-small-cell lung cancer. MATERIAL AND METHODS: Between 1985 and 1999, 77 sleeve lobectomies for bronchogenic carcinoma were performed at the University hospitals Leuven. The most common performed sleeve lobectomy is the right upper lobe sleeve lobectomy (67.5%). In 6 patients a combined sleeve resection of the pulmonary artery was performed. The operative mortality was 3.9%. Two patients developed a broncho pleural fistula. The five-year survival rate was 45.6%. In 5 patients, an anastomotic suture developed which required a completion pneumonectomy in 2. Thirteen patients developed local tumor recurrence. CONCLUSION: We conclude that sleeve lobectomy can be performed with an acceptable mortality and morbidity. Long term survival rate and recurrence rate are as good as after pneumonectomy. The operative mortality is lower when compared to pneumonectomy, exercise tolerance and quality of life are much better after sleeve lobectomy compared to pneumonectomy. For central tumours we believe that sleeve resection is the procedure of choice. PMID- 14743561 TI - Transmanubrial osteomuscular sparing approach for sulcus superior tumours. AB - OBJECTIVE: The authors report their initial experience with the transmanubrial osteomuscular sparing approach for resection of sulcus superior tumours. The feasibility of this technique is evaluated. PATIENTS: Between February 2000 and March 2002 three patients with sulcus superior tumours were surgically treated using the transmanubrial osteomuscular sparing approach. The first two patients had a non-small cell carcinoma of the upper lobe. In the third patient a pathological diagnosis of a plasmocytoma of the first rib was made. In two cases the first thoracic root was resected. RESULTS: In two patients a complete R0 resection was achieved. However, an additional posterolateral thoracotomy was necessary in two patients because the costovertebral angle was difficult to address. In one patient final histologic examination found microscopically positive margins. CONCLUSION: We believe that the transmanubrial osteomuscular sparing technique enables us to approach and control the subclavian vessels and brachial plexus in an oncologically responsible way and permits a radical resection of tumours invading the thoracic inlet. PMID- 14743562 TI - The need for communication skills in oncological surgery. PMID- 14743563 TI - The effect of social factors on diagnosis and treatment of breast cancer. AB - This study was made to evaluate the influence of the socio-economic status of a particular population in Northern Antwerp on the epidemiology and treatment of breast cancer. Data concerning patient characteristics, breast cancer pathology, treatment and socio-economical factors were compared with regional, national and worldwide literature. The patients in this series were generally older (61% more than 65 years) and tumour stage was more advanced (only 23% T1 tumours). Treatment therefore was more aggressive. The patients in our population had a lower level of education and lower income, and from a social point of view were more disadvantaged: 21% belonged to the social category of widow(er)s and orphans, while 3% had no social security. Thirty percent of patients in this series declared that late hospitalization depended on their social or financial situation. Preventive care and medical services should therefore be directed to socially and financially disabled classes of patients. PMID- 14743564 TI - Combined modality treatment for locally advanced breast cancer. AB - PURPOSE: To assess the value of aggressive loco-regional surgery in desperate situations with locally advanced breast cancer. METHODS: In this study there were considered 31 patients with locally advanced breast cancer who underwent surgery in a 5-year period. 10 of them received 3 cycles of chemotherapy and radiotherapy before surgical intervention; the rest of the 21 patients had systemic or local contra-indications for neo-adjuvant therapy. We describe clinical aspects and technical difficulties. Surgical intervention focused on tumour removal and lymph node dissection. Skin defect was covered with flaps according to the Mortimer Show technique. Postoperatively, the outcome was influenced in a favourable way by the use of Detralex, a micronized flavonoid; all but 2 patients received chemotherapy and locoregional radiotherapy. RESULTS: 25 patients survived free of disease; from 6 patients who suffered recurrence, 2 are still living and 4 have died. CONCLUSIONS: In some forms of locally advanced breast cancer, aggressive surgery offers improvement in the quality of life and increases survival. PMID- 14743565 TI - Simultaneous nipple and areola reconstruction: a review of 50 cases. AB - Nipple areola complex reconstruction has historically been a staged procedure, the nipple being reconstructed first by graft or local flap techniques and the areola on a later procedure to gain colour. This paper presents a one-stage combined reconstruction procedure using a local flap and immediate areola tattoo: it presents the advantage of reducing the total number of needed procedures and subsequently diminishes the total time needed to obtain a complete result. In this series of 50 consecutive combined procedures with a mean follow-up of ten months, no necrosis of reconstructed nipple was observed and a total of six patients needed minor revision surgery to correct nipple projection or fading of the tattoo. No local infection or wound complications were observed, underlying the safety of the procedure. Patient satisfaction and compliance were good because the procedure is fast and easy with minimal morbidity. PMID- 14743566 TI - Breast reduction mammaplasty. Shall we drain? AB - A prospective study of 35 consecutive patients who underwent breast reduction mammaplasty using the vertical scar technique was conducted to evaluate the safety of closed suction drainage omission following such a surgery. The mean age of the patients was 36.6 years and the mean resected weight per breast was 580 gr. There were two haematomas in this series, and no haematoma-related complication. These results are comparable with previously published series of breast reduction surgery using closed drainage, suggesting as reported in other works, that the absence of drains does not adversely affect postoperative recovery. PMID- 14743567 TI - Retroperitoneal lymph node dissection as adjuvant therapy in the treatment of non seminomatous testicular cancer. AB - OBJECTIVE: To assess the results of retroperitoneal lymph node dissection (RPLND) of residual masses in patients with disseminated non-seminomatous germ cell tumour treated with cisplatin-based chemotherapy, both in terms of extension of surgery, morbidity and survival. PATIENTS AND METHODS: Retrospectively, all patients treated for non-seminomatous germ cell tumour at the University Hospital of Antwerp were studied from January 1987 till December 1997. In patients with non-seminomatous testicular cancer more than stage I, the 'wait and see' strategy changed and patients were treated with chemotherapy. Patients were assessed at the end of chemotherapy and if a residual masses persisted, a RPLND was performed. If possible, a nerve-sparing lymphadenectomy was performed. Extension of surgery, morbidity and survival were analysed. RESULTS: Sixty patients had a non-seminomatous germ cell tumor of the testis and were analysed. The median follow-up was 78 months (range: 13-144 months). Thirteen patients with stage I disease were treated with orchiectomy only and none of these patients had recurrent disease. Forty-seven patients were treated with cisplatin-based chemotherapy. A complete response was observed in sixteen patients (34%), while 31 patients (66%) achieved a partial response and were treated with a RPLND. Fifteen patients underwent RPLND above the level of the renal trunk. In two patients malignant cells or fibrotic tissue were found above the renal trunk and bilateral. In five patients viable tumour cells were found in the region below the renal trunk. Sixteen patients underwent RPLND below the level of the renal trunk, of which nine had a unilateral resection, containing viable tumour in two patients. Operative mortality was 0%. One patient died six months after RPLND due to metastatic disease. In two patients, an important retroperitoneal bleeding occurred. Resection of adherent organs was performed in two patients. Long term sexual problems were reported by thirteen patients (65%) with bilateral lymphadenectomy versus two patients (18%) in the unilateral group. The survival of the patients treated with a RPLND was 97% and in the whole group of patients with a non-seminomatous testicular cancer 98%. CONCLUSION: RPLND has a place in the treatment of patients with non-seminomatous testicular cancer after chemotherapy in case of residual masses. Although mortality is low, morbidity is acceptable. In a limited number of patients there was a need of resection of adherent organs when a resection above the renal trunk was performed. PMID- 14743568 TI - Long-term results of high tibial osteotomy for medial osteoarthritis of the knee. AB - We reviewed retrospectively the results in patients who had undergone one hundred and four high tibial lateral osteotomies. The operations were all performed between 1985 and 1993. Each one of fifty men and forty nine women demonstrated a varus deformity of the knee with a coexistent medial osteoarthritis. Results were reviewed in 49 patients (62 knees) with an average follow-up of 10.2 years (range 6-14 years). Of the remaining 42 patients, 8 were lost to follow-up, 10 had died, and 24 were subsequently treated with total knee arthroplasty at an average 4.7 years after having had a high tibial osteotomy. Clinical results were evaluated using the Hospital for Special Surgery Score (HSS) and the Knee Society Score. Radiographs were systematically analysed to evaluate osteoarthritis and leg axis. Forty four (90 per cent) of the forty nine patients stated the results met their expectations and given the same circumstances, they would have the operation once again. In these patients the knee score results were excellent. The same patients had excellent HSS and Knee Society Scores. Five patients (10 per cent) had a poor result and twenty four patients were treated later by total knee arthroplasty because of pain. The following factors set these patients apart from those with more favorable results: previous arthroscopic debridement, obesity, lateral knee osteoarthritis, insufficient valgus correction, and an age of more than 55 years. High tibial valgus osteotomy provides good pain relief and improved function in carefully selected patients. Our results support this conclusion. PMID- 14743569 TI - Effect of mesh and its localisation on testicular flow and spermatogenesis in patients with groin hernia. AB - In this study, the long-term effects of mesh and its localisation (i.e. anterior or posterior) on testicular perfusion and testicular function were evaluated in groin hernia patients. Testicular function has been evaluated with spermiogram and testicular perfusion with colour Doppler ultrasonography. Group I: consisted of 30 posterior preperitoneal mesh repair patients Group II: consisted of 30 anterior tension-free repair patients. The operation types were randomised with a systematic sampling method. There was no statistically significant difference between pre-operative and postoperative spermiogram results for both groups. No statistically significant difference was found between the two groups in terms of Doppler flow parameters (PSV, EDV, RI and PI) for pre-operative, early and late postoperative periods. When Doppler flow parameters were compared for group I, statistically significant differences were found between pre-operative and early postoperative values. No statistically significant difference was found between pre-operative and late postoperative values. This is also true for early postoperative values versus late postoperative values. When Doppler flow parameters were compared for group II, statistically significant differences were found between pre-operative and early postoperative values. No statistically difference was found between pre-operative and late postoperative values. This is also true for early postoperative values versus late postoperative values. These results support the idea that inguinal mesh application is still a safe procedure in patients with no children or who are undergoing infertility treatment, where testicular function is important. PMID- 14743570 TI - Analysis of clinicopathological prognostic parameters in adenocarcinoma of the gastric cardia. AB - BACKGROUND: The optimal extent of resection for carcinoma of gastric cardia remains a subject of controversy. Although both total gastrectomy (TG) and proximal gastrectomy (PG) have their own advantages, similar survival rates were given for both. The main aim of this study is to investigate whether the extent of resection is an important prognostic factor in carcinoma of the gastric cardia. METHOD: Records of 60 patients with carcinoma of gastric cardia, operated on between the January 1989-January 1993 at Istanbul University, Cerrahpasa Medical Faculty Department of General Surgery, were reviewed retrospectively. The relationship between clinico-histopathological variables and 5-year survival was retrospectively analysed. RESULTS: Of the 14 clinico-histopathological variables, eight (age, local invasion, grade, lymphatic micro-invasion, depth of penetration, lymph node involvement, type of operation and stage of disease) were found to have a significant influence on survival. Among those clinico histopathological variables that influenced 5-year survival on univariate analysis, only age (p = 0.0029) and depth of tumour penetration (p = 0.008) independently affected survival. CONCLUSION: According to our results, depth of tumour penetration and age are the only variables which were found to independently affect 5-year survival. Depth of tumour penetration may serve as a potential marker for a biologically more aggressive tumour. The extent of resection (TG vs. PG) does not affect the long-term survival of the adenocarcinoma of the cardia. PMID- 14743571 TI - Primary gastrointestinal tract lymphomas. AB - AIM: To evaluate the prognostic factors and treatment modalities affecting survival in patients with primary gastrointestinal tract lymphoma. MATERIAL AND METHODS: Gastrointestinal tract lymphoma patients were retrospectively studied. Age and sex of patients, location and histopathological grade of tumour, stage of disease, extent of surgical resection and chemotherapy were evaluated as prognostic factors. Effects of clinicopathological and treatment associated factors on disease-free and overall survival were calculated in univariate and multivariate analyses. RESULTS: On univariate analysis, disease stage (p = 0.002), extent of surgical resection (p = 0.003) and chemotherapy (p = 0.001) were found to be significantly affecting overall survival and none of the studied factors were found to be related with disease-free survival. Multivariate analysis revealed that grade of the tumour (p = 0.042) and chemotherapy (p = 0.028) significantly affected disease-free survival and disease stage (p = 0.013) and chemotherapy (p = 0.0003) were independent prognostic factors for overall survival. In addition, surgery and chemotherapy in combination, significantly increased overall survival compared to surgery or chemotherapy only. CONCLUSION: Early stage patients have a better survival and it could be prolonged if surgery with no residual disease is performed; adjuvant chemotherapy provides an additional benefit. PMID- 14743572 TI - Intrabiliary rupture in liver hydatid cysts: results of 20 years' experience. AB - To assess the diagnostic tools and results of treatment of biliary rupture observed in liver cyst hydatids, clinical findings of 562 patients with hepatic hydatid disease were reviewed. Imaging techniques were not very effective to determine intrabiliary ruptures. Rates of rupture sizes determined in the patients were as follows; 22 (%24.7) large, 38 (%42.7) small, and 29 (%32.6) occult. Most frequently utilized procedures for patients with intrabiliary rupture were Roux-en-y cystojejunostomy, tube drainage + omentoplasty, sutured fistula + omentoplasty, and sutured fistula + tube drainage. Of the total 25 external biliary fistulas, 21 closed spontaneously. Of the four fistulas that did not close, one was managed by internal drainage and three by endoscopic sphincterotomy. Preoperative diagnosis of biliary rupture in liver hydatid cyst allows early planning of operation and helps the surgeon design the operative strategy. In the treatment of cases with large rupture, internal drainage may be proposed. PMID- 14743574 TI - Needle catheter jejunostomy complicated by pneumatosis intestinalis: a case report. AB - Needle catheter jejunostomy is a technique to allow enteral feeding after upper abdominal surgery. An unusual postoperative complication is pneumatosis intestinalis which can be life-threatening. A case of a 76-year-old man with pneumatosis intestinalis due to needle catheter jejunostomy, in whom diagnosis was made by CT-scan, is described. Needle catheter jejunostomy complicated by pneumatosis intestinalis needs attention and careful treatment. Removal of the catheter seems necessary, although controversy remains. PMID- 14743573 TI - Pre-operative Tc-99m-sestamibi scanning and intra-operative nuclear mapping: are they accurate in localizing parathyroid adenomas? AB - PURPOSE: Presentation of the results of Tc-99m-sestamibi imaging in the pre operative localization of parathyroid adenomas and the intra-operative localization of those lesions using a gamma detector (prospective study). PATIENTS & METHODS: Eighteen consecutive patients aged 27-75 years with primary hyperparathyroidism (PHPT) underwent Tc-99m-sestamibi scanning 1-2 hours before the operation and the presence of a single adenoma was recognized. All our patients underwent bilateral neck exploration based on pre-operative scanning and intra-operative gamma detector guidance and the adenoma was detected in the positions shown by both methods. RESULTS: In 16 patients we found a single adenoma localized in the same position shown by pre-operative scanning, while the intra-operative method accurately revealed all abnormal glands. In one of the two patients where an inaccurate pre-operative localization technique had been carried out, we performed thyroid lobectomy (the adenoma proved to be intrathyroidal), while the other one had an adenoma which was not close to the site indicated by the pre-operative scintigraphy. Serum calcium reverted to normal within a few days postoperatively. CONCLUSION: Patients with true-positive scans for single parathyroid adenoma could be eligible for minimally invasive operations since the abnormal gland is easily identified by the above-mentioned methods. PMID- 14743575 TI - [Fat emulsions in parenteral feeding: the present and the future perspectives]. PMID- 14743576 TI - Is cholesterol a conditionally essential nutrient in critically ill patients? PMID- 14743577 TI - [From cardiac rehabilitation to prevention]. PMID- 14743579 TI - The cumulative effects of Transcendental Meditation on cognitive function--a systematic review of randomised controlled trials. AB - It is claimed that regular practice of Transcendental Meditation (TM) improves cognitive function and increases intelligence. This systematic review assesses the evidence from randomised controlled trials for cumulative effects of TM on cognitive function. Searches were made of electronic databases and the collected papers and official websites of the TM organisation. Only randomised controlled trials with objective outcome measures of the cumulative effects of TM on cognitive function were included. Trials that measured only acute effects of TM, or used only neurophysiological outcome measures were excluded. 107 articles reporting the effects of TM on cognitive function were identified and 10 met the inclusion criteria. Most were excluded because they used no controls or did not randomize subjects between interventions. Of the 10 trials included, 4 reported large positive effects of TM on cognitive function, four were completely negative, and 2 were largely negative in outcome. All 4 positive trials recruited subjects from among people favourably predisposed towards TM, and used passive control procedures. The other 6 trials recruited subjects with no specific interest in TM, and 5 of them used structured control procedures. The association observed between positive outcome, subject selection procedure and control procedure suggests that the large positive effects reported in 4 trials result from an expectation effect. The claim that TM has a specific and cumulative effect on cognitive function is not supported by the evidence from randomised controlled trials. PMID- 14743578 TI - [Progress in the prevention of type 2 diabetes]. AB - Type 2 diabetes mellitus is a major health problem associated with excess morbidity and mortality. Defects in the action and/or secretion of insulin are the two major abnormalities leading to development of glucose intolerance. Any intervention in the impaired glucose tolerance phase that reduces resistance to insulin or protects the beta-cells, or both, should prevent or delay progression to diabetes. The natural history of type 2 diabetes includes a preceding period of impaired glucose tolerance (IGT)/impaired fasting glucose (IFG) which provides an opportunity for targeted intervention within large communities. As the prevalence of this metabolic disorder is rapidly increasing and current treatment fails to stabilise the disease in most patients, prevention should be considered as a key objective in the near future. Lifestyle intervention studies have consistently shown that quite modest changes can reduce the progression from IGT to diabetes by 50-60%. The Diabetes Prevention Program (DPP) randomised trial has shown that a combined program of weight loss, improvement of diet and increase of physical exercise lowers the risk for development of type 2 diabetes by 58% compared with placebo. It may, however, not be possible to translate these successful findings to larger cohorts or maintain the lifestyle changes longer term. This has lead to consideration of pharmacotherapy. Benefits have been found for metformin, acarbose and troglitazone. Treatment with metformin was less effective than lifestyle modifications, resulting in an average reduction of risk for development of type 2 diabetes by 31% compared with placebo. Similarly, acarbose in the STOP-NIDDM trial reduced the risk of developing type 2 diabetes in patients with IGT by 25%. Remarkably, cardiovascular event rates, in particular myocardial infarction, were significantly reduced when acarbose was used instead of placebo in subjects with glucose intolerance. The ACE inhibitors captopril (CAPPP) or ramipril (HOPE) and the Angiotensin-II receptor antagonist losartan (LIFE) have been shown to reduce the appearance of diabetes by one third when given to patients with hypertension. Since many hypertensive patients are insulin-resistant and have an increased risk in developing type 2 diabetes, the protective effect of these classes of antihypertensive drugs might be explained by their antiinsulin-resistance effects. PMID- 14743580 TI - Cholesterol improves the utilization of parenteral lipid emulsions. AB - Lipid emulsions have become an indispensable component of parenteral nutrition. Commercially available emulsions mostly have an identical composition of triglycerides (from plant oils) and egg-yolk phospholipids as emulsifier. Previous attempts to improve the composition of lipid emulsions have focused mainly on the triglyceride moiety. In the first fundamental modification of a lipid emulsion since their broader introduction into clinical medicine, we included free cholesterol in a lipid emulsion. We evaluated elimination and hydrolysis of triglycerides and lipid oxidation (by indirect calorimetry) in 10 healthy male normolipemic volunteers, comparing a conventional lipid emulsion (20% triglycerides) with an otherwise identical emulsion with the addition of 4 g/l free cholesterol. The rise in plasma triglycerides was mitigated during infusion of the cholesterol-enriched solution (323.8 +/- 27.5 vs. 202.0 +/- 18.9 mg.dL-1, p < 0.001), plasma half-life was reduced (41.6 +/- 5.4 vs. 29.3 +/- 5.1 min, p < 0.05), and total-body clearance was enhanced (0.96 +/- 0.1 vs. 1.52 +/- 0.2 ml.b.w.(.)min-1, p < 0.02). The rise in plasma free fatty acids (400.7 +/- 39.0 vs. 532.2 +/- 64.0 mumol.L-1; p < 0.02) and ketone bodies (beta hydroxybutyrate) (151.6 +/- 37.0 vs. 226.3 +/- 33.01 mumol.L-1; p < 0.02) was augmented. Increases in plasma insulin and glucagon were less pronounced (p < 0.05). The fall in respiratory quotient was greater and the fraction of lipid oxidation as a percentage of total energy expenditure was increased (66.2% +/- 6.0 vs. 70.9% +/- 6.3, p < 0.05) during infusion of the modified solution. No impairment of gas exchange or other side effects were observed. Taken together these results indicate that the elimination of a cholesterol-supplemented lipid emulsion is accelerated, triglyceride hydrolysis is enhanced, and lipid oxidation is augmented. Thus, addition of cholesterol to a lipid emulsion might not only present a means of providing cholesterol in parenteral nutrition but also help to reshape artificial lipid particles to a more chylomicron-resembling composition and improve lipid utilization. PMID- 14743581 TI - Changes in cholesterol and its precursors during the first days after major trauma. AB - BACKGROUND: The causes of hypocholesterolemia in the critically ill, including major trauma patients, have not yet been fully elucidated. OBJECTIVE: We tested the hypothesis that hypocholesterolemia is caused by decreased production of cholesterol precursors. DESIGN: Serum concentrations of squalene, lanosterol, and lathosterol were measured on admission, and then at 24 and 48 hours after injury using gas chromatography coupled with mass spectrometry. Serum concentrations of total low-density and high-density lipoprotein cholesterol were measured on admission and every day in the first week after injury. RESULTS: 83 consecutive patients with multiple trauma were examined. Significant drops in concentrations of lanosterol and lathosterol were found in the patients in comparison with the control group. The most profound drop was in lathosterol. CONCLUSION: Decreased synthesis of cholesterol precursors is the major cause of hypocholesterolemia in patients with multiple trauma. Lathosterol concentration is proposed as a marker of cholesterol synthesis. PMID- 14743582 TI - [Cardiac rehabilitation in Austria. A need to treat analysis]. AB - The aim of this analysis is to survey the general demand and current supply of cardiac rehabilitation in Austria on the basis of best evidence practice and to produce recommendations for a cost-effective structure of the entire cardiac rehabilitation system. Following the standards of indication of the Austrian Society of Cardiology an analysis of demand of cardiac rehabilitation has been carried out and juxtaposed with the current supply of facilities for cardiac rehabilitation. According to hospitalizations in the year 2000, 11,630 patients per annum would require inpatient phase II rehabilitation, 6,270 patients institutional based outpatient phase II rehabilitation and 14,319 patients institutional based phase III rehabilitation. In the year 2000, 14,746 patients received treatment in the 9 Austrian inpatient cardiac rehabilitation centres. This number is compared with an annual demand of 11,630 admissions for phase-II treatment. It follows that an equilibrium can be argued for the supply of and demand for inpatient cardiac rehabilitation in Austria. At present, 10 approved institutions in Austria offer outpatient cardiac rehabilitation services. The maximum number of positions for treatment per institution is currently 200-250. Consequently, maximally 2,000-2,500 patients per annum can be treated. In comparison, there exists a calculated demand for 6,270 patients in institutional based outpatient phase II rehabilitation and 14,319 patients in institutional based phase III rehabilitation. Altogether this amounts to a demand for 20,588 positions for treatment per annum. In Austria, the expenditures for inpatient phase II rehabilitation of a patient given an average duration of stay of 28 days, are [symbol: see text] 4,774.-. Presuming 100% compliance, the institutional based outpatient phase II rehabilitation program costs [symbol: see text] 2,760.- per patient. The costs for institutional based phase III rehabilitation services are [symbol: see text] 2,990.- per patient. This number is accompanied by a potential effective reduction of risks for the patients and a potential effective reduction of costs for the carrier as the number of rehospitalizations and recurrent procedures would decrease significantly. At present, the supply of cardiac rehabilitation in Austria is sufficient for inpatient phase II, but insufficient for the institutional based outpatient phase II and mainly phase III. Thus, a striking asymmetry exists between supply and demand. In view of the enduring effects of institutional based phase III rehabilitation, the individual and social use and finally the expected efficiency in terms of costs, this program should at least be offered without limits to all eligible patients. PMID- 14743583 TI - Long-term follow-up after renal artery stenting. AB - BACKGROUND: Renal artery stenosis may cause secondary arterial hypertension and lead to end-stage renal disease. Percutaneous transluminal angioplasty with stent implantation (PTRAS) allows effective and consistent treatment with a high technical success rate. The present trial focuses on the morphological and clinical results as assessed at a long-term follow-up (FU) visit. The main goals were assessment of the restenosis rate and evaluation of arterial hypertension and renal function. PATIENTS AND METHODS: 40 patients who had undergone successful stenting of a main renal artery were prospectively enrolled. At the FU visit, all patients underwent a risk-factor assessment, evaluation of arterial blood pressure and serum creatinine, and multi-detector computed tomography angiography (CTA). RESULTS: Median FU was 3.3 years. Hyperlipidemia was present in 67.5% of the patients, current cigarette smoking in 35% and diabetes mellitus in 15%. All patients still suffered from arterial hypertension but, compared with the pre-interventional situation, arterial hypertension was improved in 37.5%. Serum creatinine was increased in 25% of patients, mean creatinine level was 1.3 +/- 0.4 mg/dl. Hemodynamically relevant restenosis was detected by observer 1 in five patients and by observer 2 in six patients, giving restenosis rates of 12.5% and 15%, respectively, after the median FU period. Both observers detected three additional relevant stenoses in the contralateral main renal arteries. CONCLUSIONS: PTRAS gives excellent morphological long-term results. However, the clinical long-term outcome regarding arterial hypertension and renal function is only moderate. PMID- 14743584 TI - Acute hemorrhagic respiratory failure caused by Wegener's granulomatosis successfully treated by bronchoalveolar lavage with diluted surfactant. AB - Wegener's granulomatosis (WG) is an idiopathic inflammatory systemic disease that can occasionally cause an acute respiratory distress syndrome. We report on a 17 year-old girl with Wegener's granulomatosis and acute hemorrhagic respiratory failure successfully treated using bronchoalveolar lavage with diluted porcine surfactant (Curosurf; 4.8 mg/mL) followed by a low-dose bolus of surfactant. The cumulative dose of surfactant was 40 mg/kg BW. The lavage with diluted surfactant and the administration of the bolus were performed with a flexible bronchoscope. The patient was ventilated during the whole procedure, stayed hemodynamically stable and showed only a very short phase of desaturation. The PaO2/FiO2 ratio increased from 54.8 to 62.4 after one hour, to 106 after 17 hours and finally to 280 after four days. The patient was extubated five days after lavage treatment, and almost normal lung function was restored after eight weeks. Bronchoalveolar lavage with diluted surfactant by flexible bronchoscopy allows selective and direct drug administration and removes airway and alveolar debris. The technique reduces the amount of surfactant needed to overcome inhibition and thereby reduces therapy costs. We conclude that this early therapeutic intervention with surfactant might help to avoid an invasive rescue therapy such as extra corporeal membrane oxygenation, thus improving outcome in terms of faster recovery of lung function. PMID- 14743585 TI - Principalism at the bed-side. AB - Using an actual case, this paper examines a number of ways in which physicians deal with such a case and with the various "principles" and ethical theories to which they are apt to appeal. It goes on to suggest that using Dewey's method of solving problems is most applicable at the bed-side. PMID- 14743586 TI - EMTALA requirements of provider-based facilities. PMID- 14743587 TI - Florida Board of Dentistry shows its teeth to practice management companies. PMID- 14743588 TI - Provider boycotts--a review of the Merck-Medco decision. Merck-Medco Managed Care L.L.C. v. Rite Aid Corporation. PMID- 14743589 TI - Court holds joint collaboration relationship between two hospitals per se illegal. State of New York v. St. Francis Hospital, Vassar Brothers Hospital and Mid-Hudson Health. PMID- 14743590 TI - [Assessment of motor impairment with electromyography--the kinesiological EMG]. AB - Kinesiological electromyography means the application of the surface EMG measurement technique during movement and locomotion. The authors investigated the motor impairments of the upper limb of patients with spastic hemiparesis. Surface EMG activity in musculus biceps brachii and musculus triceps brachii were recorded during voluntary elbow flexion and extension movements. In some cases even the raw EMG signals contain the accurate information but more often the recorded signals should further be processed to achieve more valuable data concerning muscle performance. The limitations of the technique underlines the importance of the several steps of the processing. The purpose of this study is to present a systematic description of the methods and the precise application of surface EMG. The authors give a comprehensive view of different processing methods and demonstrate the application areas of the kinesiological EMG. PMID- 14743591 TI - [Physical and electrophysiological aspects of electroconvulsive therapy- literature review]. AB - Personnel and technical conditions for ECT (electroconvulsive therapy) is legally determined. In this article the author reviews studies focused on the technical questions of ECT. After the comparison of the unilateral and bilateral electrode placement an analysis is given on the physical properties of the stimulus used to elicit seizures. After discussing the questions of dose, wave-form frequency and polarity, the anatomical and physiological factors of impedance are evaluated. The author then discuss gender, age and encephalopathy as factors influencing seizure threshold. The connection between symmetry, regularity, and postictal suppression registered on the EEG and efficacy is observed. The EEG and EMG concordance is supposed to be connected also with the efficacy. In the discussion the author gives recommendations based on the literature. PMID- 14743592 TI - [Serotonin dysfunctions in the background of the seven deadly sins]. AB - The symbolic characters of the Seven Deadly Sins can be traced from time to time in the cultural history of human mankind, being directly specified in certain artistic products. Such are, among others, the painting entitled "The Seven Deadly Sins and the Four Lost Things" by Hieronymus Bosch and the poems Divina Commedia and The Foerie Queene by Dante Alighieri and Edmund Spenser, respectively. However, there are several paragraphs referring to these behaviours of the Seven Deadly Sins in the Bible and in the dramas of William Shakespeare. The objective of the present review is to propose that dysfunctions in the central serotonergic system might be involved in the neurobiology of these 'sinful' behaviour patterns. Evidences indicate that behaviour traits such as Accidia (Sloth), Luxuria (Lust, Lechery), Superbia (Pride), Ira (Wrath, Anger), Invidia (Envy), Avaritia (Greed, Avarice), and Gula (Gluttony) can relate to the functional alterations of serotonin in the brain. Results of biochemical and molecular genetic (polymorphism) studies on the human serotonergic system (receptor, transporter, enzyme), findings of functional imaging techniques, effects of depletion (or supplementation) of the serotonin precursor tryptophan, data of challenge probe investigations directed to testing central serotonergic functions, alterations in the peripheral serotonin measures (platelet), and the changes in the CSF 5-hydroxy-indoleacetic acid content indicate such serotonergic involvement. Furthermore, results of animal experiments on behaviour change (aggressive, dominant or submissive, appetite, alcohol preference) attributed to serotonin status modification and the clinically evidenced therapeutic efficacy of pharmacological interventions, based on the modulation and perturbation of the serotonergic system (e.g. selective serotonin reuptake inhibitors), in treating the 'sinful' behaviour forms and analogous pathological states reaching the severity of psychiatric disorders (depression, sexual disturbances, social phobia, impulsivity-aggression, obsessive-compulsive and related spectrum disorders, paranoid jealousy, eating disorders) all strongly suggest the possibility that brain serotonin dysfunctions might underlie the neurophysiology of the Seven Deadly Sins. PMID- 14743593 TI - [Risk factors for fatal outcome in subdural hemorrhage]. AB - BACKGROUND: Subdural haemorrhage (SDH) is of high public health importance because of its frequency, high case fatality ratio (CFR) and the young age of affected population. Despite the fact that the effectivity of guideline based treatment has been improved in the last decade, the Hungarian praxis shows variable compliance for recommendations. OBJECTIVES: The study aimed to describe the heterogeneity of the treatment effectivity (by geographically identifying the populations provided with appropriate or non-optimal level care), to determine the relationship between the institutional proxies quality and the results of treatment for SDH by linking the proxies properties to the patients' records. METHODS: The institutions' protocols were assessed by a self-completed questionnaire in 1997. The participating hospitals treated 79% of the Hungarian patients with SDH. The Hungarian hospital discharge data in 1997-1999 were the source of patient specific data. The risk factors of lethal outcome were investigated by logistic regression analysis. RESULTS: High proportion of patients had been treated in hospital with low compliance for guidelines. The non permanent access to neurosurgical service and CT facility, the lack of intracranial pressure monitoring and the respiration support provided out of intensive care units worsened the survival of subjects. It was quantified that the full compliance could have diminished the case fatality ratio by 15-20%. The ratio of extreme county level CFRs exceeded 2.36 and extrapolating the effectivity observed in the county with lowest lethality, the Hungarian CFR would have been reduced by 21% among patients with SDH main diagnosis. (The interpretation of findings is limited by the lack of differentiation between acute and chronic cases and of direct categorisation of severity for subdural haemorrhage in the official hospital discharge records). DISCUSSION: The study results urge the increase of compliance for evidence based guidelines, since despite of some validity issues, it was demonstrated that the deviation from recommended practice is reflected in the disadvantageous outcome. PMID- 14743594 TI - [Neurologic aspects of HIV infections--follow-up of pediatric patients]. AB - OBJECTIVES: Before the widespread introduction of combined antiretroviral therapy (1995) complications from HIV and AIDS in the central nervous system had been reported in larger proportion in infants and children than in adults: 80-90% versus 60-70%. Particular clinical manifestations tend to occur at different stages during the evolution of HIV infection. The authors review the neurological aspects of HIV infection. METHOD: First, a summary of the protocol of the neurological examinations and related experience is given. Then authors present the evaluation of neuro-psychological development, prevalence of neurological impairment and neuro-imaging of nine HIV infected children (seven boys, two girls) for the period of ten years (1991-2001). Three/ten children had vertically transmitted HIV six/nine were infected by a nosocomial route in their early childhood. Children were regularly followed up from the diagnosis of HIV. The median follow up time has been 79 month (range: 18-144 month). Four patients died during the study period. The neurological status, the motor and mental development were examined at three month intervals or monthly under one year of age. EEG was performed every six month and CT/MRI once a year. All patients received combined antiretroviral treatment and immunoglobulin therapy continuously. RESULTS: Three/nine children have normal development, one/nine has hyperactive and attention deficit disorder with normal IQ range, two/nine have slight, one/nine moderate and two/nine serious mental retardation. Mild neurological signs were found in two children, various moderate and serious neuro/psychological symptoms were found in four patients, one of them was treated with benign epilepsy too. There was also dose correlation between the clinical symptoms and the results of EEG examination (diffuse background slowing) and results of neuroimaging studies (cortical atrophy, calcification of the basal ganglia, toxoplasma abscesses). According to the results of different examinations three/nine children were found to be symptom-free, one/nine case showed the static form, two/nine patients showed the plateau form, two/nine the rapid progressive form and one/nine the progressive infantile form of AIDS encephalopathy. The majority of the patients suffered from adopting problems and difficulties of socialisation since their families lives were damaged by isolation and rejection from the community. CONCLUSION: The regular neurological and psychological examinations completed with EEG, CT/MRI were very informative to follow the course of neuro-psychological problems of HIV infected children. Symptom-free patients have to face psychosocial problems too, which cause much more damage in their mental progress than HIV itself. PMID- 14743595 TI - [Thallium poisoning induced polyneuropathy--clinical and electrophysiological data]. AB - INTRODUCTION: The aim of the study was the electrophysiological investigation of thallium induced polyneuropathy. Beyond the rarity of the illness, the motivation of this work was the possibility of following up the pattern of neuronal damage. Thallium is one of the most toxic heavy metal and its wide use increases the chance of chronic or accidental acute poisoning. The entero-hepatic circulation makes the accumulation of this toxic agent in tissues possible, mostly in neurons, in the epithelial cells of the digestive tract, in the germinative cells of the skin and testicles. In addition to alopecia and digestive complaints, the clinical picture of thallium poisoning is dominated by neurological signs. Severe axonal polyneuropathy develops in almost all cases, with further damage to the retina and impairment of cognitive functions being not unusual. The diagnosis is confirmed by finding high levels of thallium in body fluids, especially in saliva and urine. CASE REPORT: Electrophysiological examination of our accidentally poisoned patient revealed severe, sensory-motor, predominant motor axonal polyneuropathy and pointed out some aspects of the pattern of neurotoxic process: the initially distal lesion, the dying-back course and the capacity for regeneration. CONCLUSION: Because thallium has the same molecular targets as potassium ion thus impairing the energetical supply of the nerve cell, the most effective treatment is carefully loading with potassium. If recognized and treated early, thallium poisoning has a favourable prognosis. PMID- 14743596 TI - [Occipital sinus as a potential differential diagnosis in patients suspect of dural sinus thrombosis--a case report]. AB - INTRODUCTIONS: The diagnosis of dural sinus thrombosis despite of using modern imaging techniques still remains a difficult problem. For satisfactory interpretation of CT and MR scans it is indispensably important the knowledge of anatomical variations and possible imaging artifacts. One of the relatively rare developmental variations--giving chance for making false positive diagnosis--is the occipital sinus with hypoplasia or agenesis of transverse sinus. CASE REPORTS CONCLUSION: The incidence, the anatomical classification, the importance and the possibility of differentiation from sinus thrombosis are reviewed based on case reports of four patients. PMID- 14743597 TI - [The role of investigations by Janos Szentagothai in developmental neurology]. AB - The vestibulospinal system plays determining role in the activation processes of elementary sensorymotor patterns characterised by the verticalization of the trunk and elevation of the head. In the thirties of the last century Janos Szentagothai proved that axons of the vestibulospinal tract reach the cervical and thoracic spinal cord and innervate the muscles of the neck. Later he verified existence of various connections among the labyrinth, the vestibular system, and the motor nuclei of the III., IV. and the VI. cranial nerves. His studies explain the functional neuroanatomic background of sitting up, sitting and balancing in the air, head-elevation and head control during the execution of a special elementary sensorymotor pattern: "sitting in air". All these functions can be activated by labyrinthine stimulation long before the maturation of the corticospinal tract. PMID- 14743598 TI - Perspectives. Health person of the year. PMID- 14743599 TI - [Priorities in fundamental research in gerontology: Russia contribution]. AB - Current state-of-art and tendencies in the world gerontology and most important results of Russian researchers in the field are presented in the analytical review. Among the top achievements of the gerontology are: the identification of longevity genes in some animal species; Werner's progeria gene cloning; studies on genetic determinants of human longevity; discovery on evolution origin of species life span and the trade off with the reproduction; the theory of marginotomy and identification of the role of telomere and telomerase in cellular senescence; significant increase in the replicative life span of transfected with hTERT cells in vitro; the development of the free radical theory of aging; the evidence of the role of reactive oxygen species in DNA damage; the evidence of life span extension in genetically modified (transgenic, knockout or mutant) animals; the studies on the role of somatic mutations and DNA repair in mechanisms of aging, apoptosis and malignant transformation; the evidence of the life span extension by the exposure to calorie restriction diet from worms to primates and discovery of key mechanisms of the phenomena; the studies on the role of the pineal gland in the aging and the evidence of geroprotective activity of pineal peptides and melatonin. The prospects of the development of basic research on aging in Russia are reviewed as well. PMID- 14743600 TI - [Redusome aging: commentaries]. AB - The redusome hypothesis of aging and biological age control (Olovnikov, Biochemistry (Moscow) 2003, vol. 68, pp. 2-33; http://protein.bio.msu.su/biokhimiya/contents/v68/ToC6801.htm.) is discussed. Though the main part of telomere-related predictions (Olovnikov, 1971, 1973) have successfully been confirmed (end under-replication of linear DNA molecules; explanation why bacterial genome is circled to avoid this problem; telomerase existence in sex and cancer cells; correlation of telomera shortening with the number of cell doublings already performed by somatic cells that divide and age in vitro), I state that telomere model of cell aging should be abandoned, since a telomere-dependent signal of cellular senescence does not exist. Instead, it is postulated that so called redusomes are involved in control of biological time and aging. Redusomes are postulated nuclear organelles which are presented by small linear double helix DNA molecules of different specificities which are covered by proteins and located at special chromosomal nests. Each redusome has its own ori for replication, as well as promoter for transcription, but it has no centromere. Hence redusomes are distributing in mitoses among daughter cells only due to the behavior of chromosomes as their specific carriers. Transcripts from redusomes (both micro RNAs and so called fountain RNAs) participate in chromatin remodeling and chromosomal structural genes expression. Regular and consecutive losses of repeated genes from chronomeres (DNA of redusomes of neuroendoclinal and neurotrophic cells of a brain) are perceived by cells of brain's biochronometer as a course of biological time. Continuation of shortening of redusomal DNA molecules in the organism that has already achieved its physiological maturity is responsible both for cellular senescence and the organism aging. Telomere attrition is only a bystander process of aging, while the genuine cause of the cell and organism aging is the redusome DNA shortening. PMID- 14743601 TI - How the analysis of genetic mutations can help us to solve basic problems in gerontology? II. Life extending genetic modifications in budding yeast S. cereviseae, fruit fly D. melanogaster and laboratory mice M. musculus. AB - Most studies of aging are conducted in humans and domestic or laboratory animals, i.e. in conditions where artificial environment protection is applied, This yields changes in physiology and behavior, which set up organism's state unobserved in wild life. This state may be less adequate to the evolutionary adjusted genetic construction of an organism, which generates a hypothesis that in natural niches the aging rate can be lower and stress resistance can be higher than in captivity despite the fact that life expectancy in habitat is essentially lower than that in laboratory conditions due to high external mortality. Direct test of this hypothesis is difficult because of problems related to reconstruction of natural environment conditions in the laboratory. Substantial life-extending effect of some mutated genes can serve as indirect test of the hypothesis. We propose that in some cases genetic mutations can distort reaction of an organism on environmental cues and change control parameters of its life cycle. As a result such mutants in laboratory may partly demonstrate life traits similar to those observed in natural environment, e.g. associated with high stress resistance and low rate of aging. These features combined with low external mortality in laboratory conditions may lead to significant extension in the life span of mutants. Recently we considered 56 life-extending gene modifications in nematode C. elegans (Adv. Gerontol., 2003, Vol. 11), scattered in many publications. In this paper we consider pertinent life-extending gene modifications corresponded to the budding yeast S. cerevisease (29 genes), fruit fly D. melanogaster (22 genes) and laboratory mice M. musculus (8 genes). PMID- 14743602 TI - [Biochemistry of lifespan extension]. AB - A review of biochemical mechanisms underlying the known approaches to extension of lifespan and/or slowing down of ageing suggests that they all modify balances between generation of active oxygen and carbonyl species and the mechanisms that protect from their damaging effects or repair their consequences. A likely common target of the geroprotector effects of antioxidants, melatonin, and antidiabetic biguanides is the mitochondrial respiratory chain. In biological species that evolved through r-selection (nematodes, fruit flies, mice, and rats), the balance between anabolic/reproductive and self-maintenance/protective functions is the most significant modifiable factor of longevity and ageing. At the molecular level, the pivot of this balance is formed by the forkhead-type transfactors, whereas at the physiological level the balance is determined by dietary calories and physical activity via mechanism in which the central role is played by insuline-like peptides and, also, growth hormone and leptin or their functional analogs. In biological species that evolved through K-selection (higher primates, particularly humans), the latter balance is less important, and the biochemical factors of aging are more refracted through the higher regulatory systems, of which the most significant are catecholaminergic mechanisms of regulation of neuroendocrine-immune interrelationships and the circulatory system. This results in a decreased geroprotector potential of calory restriction and in an increased importance of the optimal physical activity. When these conclusions are compared with demographic data, it comes out that virtually all advances in gerontology may be reduced to maxims of healthy ageing known from extreme antiquity. Under optimal socioeconomic conditions, the chances to approach the documented world record of human longevity (122 years) may be increased by (not to mention getting rid of smoking and other abuses) high physical activity, adequate nutrition enriched in fresh fruits, optimism, and timely treatment of specific diseases. The most important bottleneck in the realization of these reserves is currently the public consciousness rather than the science. PMID- 14743603 TI - [Polymorphism of the gene of angiotensin-converting enzyme and apolipoprotein E gene in long-livers of Novosibirsk city]. AB - The levels of polymorphism of genes of angiotensin converting enzyme (ACE) and apolipoprotein E (Apo E) were studied in elderly and long-living people in Novosibirsk. The results of the study in the investigated group (97 subjects) were compared with polymorphism of these genes in Novosibirsk population group aged 25-64 who were investigated in MONICA Project survey and had DNA data base formed. Frequency of D/D genotype among senile and long-living men was 5.9%. It is 5 times lower than in men 55-64 years of age (p = 0.04). Similar decrease of this gene frequency was also found in women of the same age. In men older than 83 years of age 4 times lowering of 3/4 genotype of Apo E gene and 2 times increasing of frequency of 2/3 genotype were revealed when comprising frequency of these genotypes in people of middle age. In subjects of senile age and long livers of both sexes genotype 4/4 was not revealed. Lipid levels were more favorable in women with genotype 2/3 of Apo E gene (comparatively lower mean level of total cholesterol and higher level of HDL cholesterol) if compared with genotypes 3/3 and 3/4. PMID- 14743604 TI - [Longevity in Saint-Petersburg: basic medical problems and requirement for a medical care]. AB - The aim of the study was to analyze a demographic situation in St. Petersburg with a special attention to the oldest old. The proportion of long-living persons in total population is increasing in all developed countries. The rate of elderly people grows rapidly alongside with the decrease of the birth rate. The number of the oldest people (90+) has grown especially rapidly during the last decade--from 5.9 to 11 thousands in St. Petersburg. Frequency of visits to general practitional, specialist, and emergency for medical aid during 1996-2002 have been studied in the city. Special groups of centenarians have been selected. 396 persons over 85+ years were under observation in the basic group. There were 38 centenarians among them. Seventy-one person of 40-55 year-old and 110 persons of 65-75 year-old were included in the control group. Elderly people have specific medical problems: vascular or Alzheimer-type dementia, osteoporoses and its complications, diabetes, incontinence, diseases of sense organs. The incidence of an appeal to the urgent medical aid was higher among persons of control group than that in the 85+ group. There were more cases of chronic pathology and less cases of acute diseases in the oldest group as compared with young ones. In centenarians, address for different kinds of a medical care is reduced in comparison with other age groups of long-livers. There were no registered oncological pathologies and "disappearing" of some chronic diseases have been observed in centenarians. CONCLUSIONS: Our study prove the necessity of the creation of medical and social geriatric services in the city. The improve of the qualifications of geriatricians is an important task. PMID- 14743605 TI - [Immune system senescence and mortality dynamics. Role of antigenic load]. AB - This paper is dedicated to investigation of the impact of total antigenic load on immunosenescence. Data on pneumonia mortality have been simulated. The results of modelling revealed that antigenic load as well as probability of infecting could account for the shifting in mortality that takes place for different countries. Dynamics of immunosenescence is described by mathematical model for age related changes in T-cell populations. Numerical simulation of the model yields estimates for pneumonia severity at different ages. Probability function is used to describe relationship between pneumonia mortality and age related changes in disease severity. PMID- 14743606 TI - [Role of free-radical processes in age-dependent change of energy provision in the liver of rats in stress]. AB - This study was designed to assay the effect of dimethyl sulfoxide to adenine nucleotide and inorganic phosphate concentration in the liver of adult (10-12 month) and old (22-25 month) Wistar male rats during immobilization stress. Investigation have shown that dimethyl sulfoxide injection prevented of AMP concentration decreasing and has no effect to magnitude of ATP decreasing in the both adult and old rats during stress. This antioxidant prevents reduction of total adenine nucleotides level in the liver of old rats during immobilization also. The role of free radical processes stimulation in the adenylate system changes in the liver of adult and old rats during stress have been discussed. PMID- 14743607 TI - [Age-dependent reconstruction of element status in Magadan inhabitants]. AB - On the base of macro- and microelements' concentration in hair of Magadan's inhabitants there were analyzed the changes of aged people's element status. It has been shown that with age, differently directed reconstruction happened, leading to accumulation of one elements and decrease of concentration of others in inhabitants' hair. Elements, undergoing natural age reconstruction, may be considered as indicators and initiators of aging. We have allocated two groups of elements, in dependence on changes' direction. The first group is the main one; Na, K, Ca, Se, As, Mg, Mn, Fe, Zn belong to this group; their level in hair is increasing with age. Taking into account the excretory function of hair, accumulation of named elements is considered as aged removing of these elements out of organism. The exception is the toxic As: its superfluous concentrations in hair reflect the accumulation As in the internal environment of organism. Elements (Cr, Si), concentration of which in hair is decreasing with age, belong to the second group. On the base of literary data, the decreasing of named elements in hair must be interpreted as their programmed decreasing in the organism of aged people. Hence, in aged people there is seen the generalized decreasing of the most studied essential macro- and microelements in the organism. The exception is As, concentration of which is increasing in aged people. The age disorder of element homeostasis may serve as a predictor of "normal" diseases and may be one of pathophysiological mechanisms of aging. PMID- 14743608 TI - [Regulating effect of pineal gland peptides on development of T-lymphocytes in CBA aging mice: role of microenvironment of immune system organs and neuroendocrine factors]. AB - Studies were undertaken on the development of T-lymphocytes in adult and old CBA mice and its changes at aging after injections of pineal gland peptides. It was shown that in old mice the disturbances of T-cells differentiation are registered in bone marrow, thymus, spleen and characterized by the changes of lymphocyte markers expression, migration and proliferation of cells. In old mice FTS titer, melatonin and testosterone levels decreased, the balance of noradrenalin and serotonin in hypothalamus and the cell composition of microenvironment immune systems organs impaired. After chronic (18 mo) administration of the pineal gland preparation epithalamin the amount of stromal cells-precursors, CD4+ and Mac-1(+) cells in old bone marrow increased, improved the migration of T-cell precursors from bone marrow to thymus and their proliferative potential. The proportion of CD3+, CD4+CD8-, CD4-CD8+, Mac-1(+)-cells in old thymus increased, while that of CD44(+)-cells decreased. The proportion of CD4-CD8(+)-cells in spleen increased. The most number of indices and their balance showed a pattern of adult mice. In old mice after epithalamin the balance of amines in hypothalamus improved, concentration of melatonin in pineal gland, testosterone and FTS titer in blood increased. Epithalon has also the possibility to increase of thymic endocrine function. PMID- 14743610 TI - [Combined effect of vilon and cyclophosphane on tumor transplants and lymphoid tissue explants in mice and rats of various age]. AB - The experiments were performed in mice with transplanted Lewis lung carcinoma. The injections of synthetic peptide vilon at the doses 1 mg/kg significantly increased the survival of mice. So vilon has possessed the oncomodulating action on the transplanted carcinoma. The synchronous injection both of vilon and cyclophosphan at the doses 100 mg/kg decreases the survival of mice. There was also studied in spleen organotypic tissue culture the effect of vilon and cyclophosphan on the development of explants of rats of various age: 1 day and 2 years old. Vilon stimulated apoptosis both in young and old rats, but the inhibitory effect of cyclophosphan was abolished in the presence of vilon in culture media at the dose 5 ng/ml. The results obtained suggested that perspective preparates in the cancer therapy--vilon and cytostatic drugs-, must be used not synchronously. PMID- 14743609 TI - [Peptide correction of age-related pineal disturbances in monkeys]. AB - Investigation of the age-related changes of the pineal gland function and possible ways for their overcoming on nonhuman monkey model was the purpose of this study. Hormonal function of the pineal gland was studied in 38 Macaca mulatta females of two age groups: 6-8 years old, n = 18 and 20-26 years old, n = 20. Pineal function was studied in basal conditions and after administration of pineal peptide preparations--epithalamin and epitalon, both developed in the St. Petersburg Institute of Bioregulation and Gerontology (Russia). It has been revealed that plasma melatonin concentration in monkeys has well expressed high amplitude diurnal rhythm. Minimum is manifested at 4 p.m. and maximum--at 10 p.m. 3 a.m. In aging the mean diurnal melatonin concentration decreases by 1.5-2 times as well as in different points of the day: 9 p.m., 10 p.m., 3 a.m. and 4 a.m. Administration of pineal peptides--epithalamin (at the dose 5 mg/animal/day intramuscularly during 10 consecutive days) or epitalon (at the dose 10 micrograms/animal/day intramuscularly during 7-10 consecutive days) induced significant increase in the night plasma melatonin in old monkeys, but the treatment did not change the melatonin level in young monkeys. Taking into consideration that melatonin is very important for regulation of the diurnal rhythm of functioning of some organs and systems it should be suggested that applying epithalamin and epitalon are perspective in the correction of age related hormonal imbalance and age pathology. PMID- 14743611 TI - [Melatonin for diagnosis of cancer and assessment of prognosis in elderly patients]. AB - A review is given on the analysis of melatonin (MT) and of its main peripheral metabolite 6-sulfatoxymelatonin (aMT6s) in old patients with different types of primary unoperated carcinomas. A very low production of melatonin (as estimated by the nocturnal urinary excretion of aMT6s) was found in male patients with lung or stomach cancer compared to aged-matched controls as well as in female patients with thyroid cancer. The levels of these women, however, did not differ from female patients with benign thyroid diseases indicating a general suppressive effect of thyroid diseases on the pineal gland. A similar but opposite phenomenon was observed in male patients with primary unoperatide colorectal cancer which showed an elevated production of mellatonin when compared to healthy men but not when compared to patients with colitis ulcerosa. The mechanisms involved in these phenomena are poorly understood and seem to include central as well as peripheral components. This view is supported by the finding that in spite of varying urinary aMT6s excretion measured in patients with different types of tumor, aMT6s shows comparable positive correlations with the degree of tumor cell proliferation (as estimated by the number of PCNA-immunopositive cells). Therefore the amount of aMT6s excreted (as well as the corresponding concentration of circulating MT) has to be understood as the net result of a number of different effects by the tumor on organism. PMID- 14743612 TI - [Prematurely aging OXYS rats as an animal model of senile cataract in human]. AB - Cataracts remain the major cause of blindness and acuity of vision deterioration in the elderly. To date there are no effective methods of preventive measures of this disease and surgical removal of cataract-damaged lenses remains the only way to restore vision. To explore cataract pathogenesis and development methods of its prophylaxis are necessary fundamental investigation, which is impossible without biological models. As the first Russian her senile cataract model it is suggested to use the senescence-accelerated OXYS rat strain which was developed 30 years ago by selection and inbreeding of rats Wistar susceptible to cataractogenic effect of galactose (Institute of Cytology and Genetics, Novosibirsk). We conducted strong selection by the early cataract during 13 generations of OXYS rats. As the result at present time changes of OXYS rat lens appear by the 2 month, at 6 months these changes appear in 100% of animals (in rats Wistar--5%), at 12 month cataract affects both eyes. The data suggest that the cataract in rats OXYS develops simultaneously with progressive macular degeneration while changes of the eye lenses morphologically correspond to senile cataract in man. PMID- 14743613 TI - [Mathematical modeling in gerontology--strategic perspectives]. AB - Mathematical description of aging and mortality is presented. A review of tendencies in modeling is discussed and some difficulties related to adequacy and the ways of information presentation in the models are talked about. A short view is given at the two main modeling techniques in biology, namely the models of data and models of a system. Strategic perspectives of modeling in biogerontology are presented in terms of modeling paradigms. One can speak about a general paradigm consisting of postulates, principles and techniques related to a model, which allows a solving different problems with the same model, if the model can be modified to treat the variety of problems. Modification is to be done in the framework of concepts of the model by using a sum of the methods related to it. It is shown how the homeostatic paradigm can be used to predict a specific human longevity. PMID- 14743614 TI - [Can a biologist fix a radio, or what I learned while studying apoptosis]. PMID- 14743615 TI - [People fix radio because they can create them]. AB - Comments to the article of Yu. Lazebnik Yu. "Can a biologist fix a radio?--Or, what I learned while studying apoptosis". PMID- 14743616 TI - [A mathematician will help a biologist to fix a radio]. AB - Comments to the article by Lazebnik Yu. Can a biologist fix a radio?--Or, what I learned while studying apoptosis. The role of mathematic modeling in biology discussed in the paper. PMID- 14743617 TI - Your professional home is strong and vigorous. PMID- 14743618 TI - Physician leadership. PMID- 14743619 TI - Medicare reform passes: physician fee cut averted. PMID- 14743620 TI - Administrative complexity. Money down the drain. PMID- 14743621 TI - Leasing information technology. Is it right for your practice? PMID- 14743622 TI - Be the performance standard. A staff operations primer. PMID- 14743623 TI - How to get--and keep--employees' trust. Good management is the foundation. PMID- 14743624 TI - Eight to the bar. Success factors affect operations, strategy. PMID- 14743625 TI - Strength in numbers. Medical group practices fill vital niche in U.S. health care system. AB - As the Medical Group Management Association initiates and celebrates National Medical Group Practice Week, Jan. 26-30, we look back at the evolution of this care delivery model in the United States. Echoes of the historical forces shaping group practice remain; modern pressures continue to influence it. Medical group practice has proven a necessary, responsive and adaptable segment of the health care delivery system. PMID- 14743626 TI - Hidden expenses. The true cost of adding a physician to your medical group practice. AB - A new physician's salary is the most visible expense associated with recruitment, but it is not the only one. A practice should anticipate two other significant direct costs and two indirect costs when it prepares to add professional staff. A physician recruitment financial model describes the true expenses that come with recruiting and prioritizes them for planning. PMID- 14743627 TI - Accept a substitute. But make sure to file claims for locum tenens services appropriately. AB - With the rapid growth of the locum tenens industry, it's increasingly important to know how to bill Medicare for locum tenens' services. The author describes when physicians and medical practices may bill Medicare for locum tenens' work and the sanctions they face for improper billing. PMID- 14743628 TI - When goodwill brings no good. IRS challenging donation of goodwill by medical groups. AB - Should physicians be allowed a charitable tax deduction for the value of their practice goodwill when their organization is acquired by a not-for-profit entity? This article explores the circumstances and ramifications of two court cases. PMID- 14743629 TI - [Expression of toll-like receptor mRNA in epithelial cell of nasal mucosa]. AB - OBJECTIVE: To understand the expression of Toll-like receptor-2 (TLR-2) and Toll like receptor-4 (TLR-4) in epithelial cell of nasal mucosa, and to investigate the role of Toll-like receptors in the innate immunity of nasal mucosa. METHODS: Specimens from 30 patients with chronic sinusitis and 21 healthy adults were examined by in situ hybridization for TLR-2 and TLR-4 mRNA. RESULTS: All 30, samples of chronic sinusitis showed a stranger expression of TLR-2 and TLR-4 than in controls (t = 8.605, P < 0.0005, t = 9.050, P < 0.0005). CONCLUSION: It is suggested that epithelial in nasal mucosa were not just a physical barrier, which could represent a novel defense mechanism of the host during bacterial infections through Toll-like receptors. PMID- 14743630 TI - [A study of the link between upper and lower respiratory inflammation]. AB - OBJECTIVE: To evaluate the correlation between upper and lower respiratory inflammation. METHODS: Thirty rats were randomly divided into allergic rhinitis group (AR), asthma group (AS), and control group (Con). The pulmonary function was measured by pulmonary function instrument of animal model, the infiltration of eosinophils and mast cells was detected by hematotoxylin-eosin staining (HE staining) and toluidine blue staining respectively, the expression of VCAM-I and IL-13 was examined by immunohistochemistry. ELISA was used to measure the concentration of IL-5 in the peripheral serum. RESULTS: The numbers of eosinophils and mast cells in nasal mucosa and lung tissue of AR were both significantly higher than those of Con, and similar results were observed between AS and Con. The forced expiratory volume in 0.3 second (FEV0.3)/forced vital capacity (FVC) of AR and AS was both significantly lower than that of Con. The numbers of VCAM-I and IL-13 positive vessels in lung tissue of AR were both significantly higher than those of Con, and similar results were observed between AS and Con. The concentration of IL-5 in the serum of AR and AS was both significantly higher than that of Con. The concentration of IL-5 in serum was positively correlated with the number of eosinophils in peripheral blood of AR and AS. CONCLUSION: The inflammation is similar between AR and AS, and repeatedly challenge upper respiratory tract can impact on lower respiratory response. PMID- 14743631 TI - [Acute fulminant invasive fungal sinusitis]. AB - OBJECTIVE: To discuss the clinical presentation, diagnostic criterion and treatment principle of acute fulminant invasive fungal rhinosinusitis (AFFS). METHODS: Six patients were diagnosed as AFFS based on history, nose, especially eye symptoms with or without fever, sinus CT and MRI, endoscopic and cytological findings in the nasal cavity. Surgical debridement was performed on 5 of 6 biopsy proven AFFS patients, one of them being amputated of the orbital content. With the original disease controlled simultaneously, 4 of 5 patients were prescribed with systemic amphotericin B or liposomal amphotericin B (1 case) and one with Itraconazole orally. One patient had not been involved with any of the anti-fungi measures. RESULT: Mucor (Zygomycetes) was identified on culture in 1 patient, Rhizopus species in 2, Aspergillus in 1, Alternaria in 1 and mixed Mucor and Rhizopus in 1. All patients were proved of tissue invasion histopathologically through biopsy. One patient died without any anti-fungi therapy on the 7th admission day, 3 patients survived for 88 days, 32 and 6 months respectively and died of original diseases (diabetes 1, leukaemia 2). One patient survived 9 months and lost for follow-up, 1 patient survived 11 months after treatment. CONCLUSION: A high index of suspicion and early endoscopic investigation through nasal cavity with fungal investigation should highly be strengthened for recognition of this disease. MRI findings should be considered as, or even more, important as that of CT scan on the early diagnoses. Extensive and aggressive surgical debridement, prompt and enough dosage of antifungal therapy intravenously, together with serious controlling of the underlying disease, all take important roles in the complete control of the disease. PMID- 14743632 TI - [Long-term outcomes and analysis of its relative factors of nasal endoscopic sinus surgery in children]. AB - OBJECTIVE: To evaluate the results of endoscopic sinus surgery (ESS) in children with chronic sinusitis. The long-term outcomes and the relative factors were analyzed retrospectively. MATERIALS AND METHODS: A total of 268 children (432 sides), 186 males and 82 females, aged 3 to 17 years (average, 14.3) were included in this study (underwent ESS). The postoperative follow-up time of 188 cases (305 sides, 70.1%) was over one year. The management during follow-up was adopted under nasal endoscope and local anesthesia. The postoperative treatment included cleaning operative cavity, nasal irrigation, antibiotics, steroid, immune regulator, etc. RESULTS: Overall cases included 89 cases (147 sides) of chronic sinusitis, 86 cases (142 sides) of nasal polyps and 13 cases (16 sides) of mucocele respectively. The cure rate was 70.2% (132 cases). 43 cases (22.9%) were improved, while other 13 cases were of no effect. Overall symptoms, 93.1% children with headache were improved, 85.1% children were free of nasal obstruction and 60.1% children gained a relief of purulent discharge. 11 cases had no change and 2 cases were worse compared with before surgery. 34 cases with anosmia were improved or recovered after ESS. The total effective rate of ESS was 93.1%. 8 cases received a revision ESS. CONCLUSIONS: ESS in children with chronic sinusitis and polyps has a good long-term outcomes. The reasonable reservation of mucosa during ESS must be emphasized. The quality of follow-up is an important factor affecting the results of ESS. Whether the disciplinary preoperative management was made or not is the hypothesis of ESS indications. PMID- 14743633 TI - [Experimental study of sodium nitroprusside on whole cell calcium currents of outer hair cells in the cochlear of guinea pigs]. AB - OBJECTIVE: To study the effects and mechanism of sodium nitroprusside (SNP), a donor of nitric oxide, on the calcium currents of isolated outer hair cells (OHCs) from the cochlear of guinea pigs. METHODS: Acute isolated outer hair cells of guinea pigs were performed, and the whole cell patch clamp recording techniques were used. The K+, Na+ ions were excepted to study effects of SNP on the calcium currents of outer hair cells. RESULTS: SNP inhibited the inward calcium currents of OHCs. Under the condition of holding at -60 mV and stimulation voltage as +10 mV, SNP (10 mmol/L) inhibited (61.12 +/- 1.99)% of the whole cell's calcium currents (n = 5). A dose-reaction response was obtained from 5-8 cells. The half inhibiting concentration was 1.9 mmol/L while the maximum inhibiting concentration was 100 mmol/L, but Hill coefficient was 0.98. SNP could selectively block the L-type calcium channels on outer hair cells (n = 6). CONCLUSION: As a donor of nitric oxide, SNP could affect the physiology function of outer hair cells by inhibition of the calcium currents by blocking the L-type calcium channels. PMID- 14743634 TI - [Effect of linopirdine on potassium currents in isolated outer hair cells and Deiters' cells of the cochlea from guinea pig]. AB - OBJECTIVE: To study the effect of linopirdine, a selective blocker of KCNQ channel family, on the whole-cell potassium currents in isolated outer hair cells (OHCs) and Deiters' cells of the cochlea from guinea pig and to probe the distribution of KCNQ channel family in OHC and Deiters' cells. METHODS: Using the patch clamp whole-cell recording, IK (tetraethylammonium-sensitive outward potassium currents) and IKa (inward potassium currents activated at negative potential) were measured before and after the administration of linopirdine in OHCs. Simultaneously, IK (the outward rectifier potassium current) was recorded in Deiters' cells. RESULTS: IK was partly reduced after the administration of linopirdine in OHCs, moreover, IKa was totally inhibited. However, IK in Deiter's cells was not decreased. CONCLUSION: KCNQ channel family could only involve in constructing the variant potassium channels in outer hair cells, but not in Deiters' cells. PMID- 14743635 TI - [Research of relationship between endoneurial fluid pressure and electroneurography of the facial nerve]. AB - OBJECTIVE: The relationship between endoneurial fluid pressure (EFP) and electroneurography (ENoG) of the facial nerve was studied in order to evaluate the ENoG as the basis of pathophysiology of the decompression of the facial nerve. METHODS: The values of ENoG were recorded by an instrument of physiology on the normal and crushed facial nerves of guinea pigs. Endoneurial fluid pressures were measured by a servo-nulling micropipette system at the same time. After the EFP were measured, the facial nerves were removed and fixed properly for examination under the light and electron microscope in order to determine the differences from various periods after injury. RESULTS: EFP in the facial nerve was changed significantly during the period of three days to three weeks later after crushed injury, and the percentage of degenerated facial nerve fibre was the same. There was positive correlation between ENoG and EFP. After crushed injury 2 and 3 weeks, the coefficient was 0.88 and 0.51, respectively. It could be found that extensive edema in endoneurial and perivascular spaces of the facial nerve occurred in early stage of injury, but numerous Schwann cells proliferation appeared in the later period. CONCLUSION: The change of ENoG could reflect the value of EFP relatively within three weeks after crushed injury. It was suggested that ENoG could be useful in evaluating the pathogenesis underlying facial palsy. And it could be provided a basis indication for decompression of the facial nerve. PMID- 14743636 TI - [Expression and distribution of aquaporin-2 in nasal polyps]. AB - OBJECTIVE: To study the distribution and expression of aquaporion-2 (water channel protein; AQP-2) in nasal polyps and to evaluate the role of AQP-2 in the formation of nasal polyps. METHODS: Eleven samples of normal inferior turbinates and forty-six samples of nasal polyps were used. In the group of nasal polyps, there were ten cases of type II phase 1, twelve type II phase 2, ten type II phase 3 and fourteen type III. The expression of AQP-2 in both groups was studied with immunohistochemical technique. RESULTS: 1. Both the number of AQP-2 positive cells and the area density in the epithelial cell layer from nasal polyps were higher than those from inferior turbinates (P < 0.01, P < 0.05 respectively); In the group of nasal polyps, the area density from type III, type II phase 2 and type II phase 3 was significantly higher than that from type II phase 1 (P < 0.05); 2. Both the number of AQP-2 positive cells and the area density in the subepithelial tissue from type III, type II phase 2 and type II phase 3 nasal polyps were higher than those from type II phase 1 nasal polyps and inferior turbinates(P < 0.05), whereas the difference between the latter two was not significant(P > 0.05). CONCLUSION: The high expression of AQP-2 protein in nasal polyps may contribute to the genesis and development of nasal polyps. PMID- 14743637 TI - [Analysis of growth model in vestibular aqueduct during human fetal development]. AB - OBJECTIVE: To characterize the growth of the developing vestibular aqueduct in order to gain a better understanding of the possible origins of enlarged vestibular aqueduct. METHODS: Serial sections from 32 temporal bones for every other week from human embryos ranging in age from 6 to 38 weeks were studied with 3DMed medical image analysis software. The internal, external aperture, midpoint diameter and length vestibular aqueduct were analyzed with regression analysis to obtain a growth curve. RESULTS: The internal aperture of vestibular aqueduct was began to form at 6 weeks' gestation. It grew to reach the posterior surface of the petrous bone in the posterior cranial fossa by 10 weeks' gestation. All width parameter mean value in our embryos nerve reached the maximum average width in the adult. Statistical analysis showed that the vestibular aqueduct grew in a nonlinear continuous fashion and instability trend throughout embryonic life, except length parameter. CONCLUSION: The vestibular aqueduct grows in a nonlinear fashion throughout embryonic life. The widest aqueduct measured in embryonic life does not reach the maximum average width in the adult. These results suggest that it would be possible for vestibular aqueduct to develop postnatally. PMID- 14743638 TI - [Pneumatization of perpendicular plate of the ethmoid bone and mucocele]. AB - OBJECTIVE: To find out the prevalence of pneumatization of perpendicular plate of the ethmoid bone, and to discuss the diagnosis and management of perpendicular plate mucocele (nasal septal mucocele). METHODS: The CT data from 32 patients with septal deviation were reviewed, and an unusual case of perpendicular plate mucocele was reported. RESULTS: Six cases (6/32, 18%) were found to have pneumatization of perpendicular plate of the ethmoid bone, with 2 located in the anterior portion (frontal-septal pneumatization), and 4 located in the posterior portion (spheno-septal pneumatization). A patient with perpendicular plate mucocele was treated by the technique of marsupialization under nasal endoscope. This patient was followed-up for 10 months without recurrence. CONCLUSION: As a kind of variation of nasal septum, the clinical significance of pneumatization of perpendicular plate should be emphasized. We reported the first case of perpendicular plate mucocele, originated possibly from the pneumatization of perpendicular plate of the ethmoid bone. Marsupialization under endoscope was considered to be the initial management for this unusual disease. PMID- 14743639 TI - [Malignant fibrous histiocytomas of larynx]. AB - OBJECTIVE: To explore the histopathology, diagnosis and treatment of malignant fibrous histiocytomas (MFH) of larynx. METHODS: The clinical and pathologic features of 9 cases with MFH of larynx were reviewed retrospectively. RESULTS: All the 9 cases with MFH of larynx were adults (the age ranged from 43 to 87 years; average 56 years); 8 were men and 1 woman. The tumor of eight cases originated in glottic area and 1 case occurred in supraglottic area. The histological diagnosis was difficult, so the immunohistochemical studies in 5 cases were examined. Because surgical resection was a mainly treatment, seven patients underwent total laryngectomy and 2 cases underwent frontal partial laryngectomy. Six patients were followed up for more than 3 years and no patient died of MFH after surgical resection. CONCLUSION: The diagnosis of MFH of larynx was sometimes difficult and immunohistochemistry could be helpful. The mainly treatment of MFH of larynx was surgical resection by total laryngectomy or partial laryngectomy. PMID- 14743640 TI - [Effects of pingyangmycin on mucous membrane of respiratory tract and its prevention]. AB - OBJECTIVE: To observe the effects of pingyangmycin(PYM) on the histomorphology and ultrastructure of airway mucosa from the healthy Wistar rats and explore the intervenient role of Vitamin E and Composite Salvia Miltiorrhiza (CSM) on these effects. METHODS: Thirty Wistar rats were divided randomly into four groups: Group A as control group (injection of N. S into peritoneal cavity); Group B received intracavitary injection of PYM; Group C received both intracavitary injection of PYM and tube feeding of Vitamin E; Group D received intracavitary injection of PYM and CSM. The nasal and tracheal mucosa were taken for light microscopy(LM), transmission electron microscopy(TEM) and scanning electron microscopy(SEM) at the 1st, 2nd, 4th and 6th week of the experiment. RESULTS: Edema and degeneration of epithelial cells manifested as expansion of endoplasmic reticula, swelling of mitochondria, disarrangement of mitochondrial crista, vesicula formation of mitochondria, reduction of mitochondria in part cells, mild expansion of perinuclear space and slight peripheral accumulation of nuclear chromatin could be seen in Group B at the end of the 1st week without necrosis or detachment of the cells. At the 2nd week, there were necrosis and exfoliation of epithelial cells, rupture of cell membrane, phenomena of outflow of mitochondria and endoplasmic reticula. The degree of cell damage became more seriously at the 4th week and was not recovered to normal condition even 2 weeks after stopping administration of PYM. The cell damages in Group C and D were significantly lighter than that in Group B and its occurrence were significantly postponed. The injured cells recovered definitely after stopping PYM administration and continuous application of Vitamin and CSM for another 2 weeks. CONCLUSIONS: (1) The nasal and tracheal mucosa of Wistar rats can be damaged by injection of PYM and the degree of damage which become more serious with the increase of dose. (2) Vitamin E and CSM can alleviate the damages of the respiratory mucosa from Wistar rats caused by PYM. Both drugs could be used to prevent respiratory tract mucosa from the damages induced by PYM in clinical. PMID- 14743641 TI - [Application of bi-pedicel and bi-muscle flaps for the expanded partial laryngectomy in the patients with T3T4 laryngeal cancer]. AB - OBJECTIVE: To evaluate the application of bi-pedicel and bi-muscle flaps for the expanded partial laryngectomy in the patients with T3T4 larynx cancer. METHODS: From 1990 to 1997, 386 patients of laryngeal cancer were treated by expanded partial laryngectomy and repaired with unilateral or bilateral bi-pedicel and bi muscle flaps. 322 patients were male and 64 were female. The age ranged from 30 to 84 years old. All the patients were in the stage T3 or T4 of cancer. The operation included expanded horizontal supraglottic laryngectomy, vertical partial laryngectomy, 3/4 laryngectomy and sub-total laryngectomy. RESULTS: After operation, the speech function was still good in all cases. The swallow function recovered in 8 to 12 days post-operation. The extraction rate of tracheal tube was 50.3% (194/386), and the 5-years survival rate was 55.2% (213/386) while the local recurrence rate was 16.6% (64/386). CONCLUSION: With the application of bi pedicel and bi-muscle flaps, the patients, if treated by total laryngectomy as traditional technique, could be treated by partial laryngectomy. This method could expand the indication of partial laryngectomy and improve the life quality and survival rate. PMID- 14743642 TI - [CO2 laser endoscopic subtotal arytenoidectomy for bilateral median vocal cord paralysis]. AB - OBJECTIVE: To delineate the surgical procedures and correlated techniques for endoscopic subtotal arytenoidectomy, as well as to discuss their applications and clinical outcomes. METHODS: CO2 Laser endoscopic unilateral arytenoidectomy was performed in eight cases of bilateral median vocal cord paralysis combined with one stage of mucosal micro-anastomosis. All patients suffered from dyspnea in some extent, of which 5 had the history of thyroidectomy and 2 had traumatic causes following esophagectomy and tracheal surgery respectively. One of patient had unknown cause. Six patients had undergone tracheotomy prior to operation or before their referral to our hospital. The airway was evaluated via fibro-optic laryngoscopy, and the voice quality was assessed subjectively by the patients and the surgeon before and after surgery. RESULTS: Following 5-43 months after the surgery, in all cases the function of airway as well as the acceptable voice quality was successfully restored. The tracheotomy done before operation in six patients was decannulated within the mean time of 44.2 days post-operation. CONCLUSION: The endoscopic approach for CO2 laser unilateral arytenoidectomy may lead to better restoration of an adequate airway and satisfying phonation without postoperative aspiration. Mucosal micro-anastomosis can prevent the formation of granulation or scar tissue thus promotes the healing processes. This procedure is simpler than other ordinary surgical methods, and could be a satisfactory alternation of treatment for bilateral median vocal cord paralysis. PMID- 14743643 TI - [Surgical management of hypopharyngeal cancer]. AB - OBJECTIVE: To investigate the preservation of laryngeal function for the patients with hypopharyngeal cancer. METHODS: Two hundred and ninety-three cases of hypopharyngeal cancer with surgical management were reviewed retrospectively, and 222 cases were originated from pyriform sinus, 13 from post-cricoid, and 21 from posterior pharyngeal wall. Radiotherapy (37 cases), operation only (56 cases) and the combined treatment (operation plus radiation or chemotherapy, 200 cases) were adopted. 159 cases were treated with function preserved laryngectomy and 97 with total laryngectomy. RESULTS: The 5 year survival rates of patient with laryngeal function preserved and no laryngeal function preserved were 51.3%, 47.6% (for stage III); 40.4%, 43.3% (for stage IV), respectively. There were no significant differences in 5-year survival between the functionally preserved group and no functioned group (P > 0.05). The analysis of survival rates revealed a significant difference between combined therapy and radiotherapy. CONCLUSION: There is no significant difference between the survive rates of function preserved and non-preserved groups. Conservation laryngectomy improves the quality of patient's life, and combined therapy is the best choice for hypopharyngeal cancer. PMID- 14743644 TI - [Clinical analysis of relevant factors causing laryngeal cancer postoperative recurrence]. AB - OBJECTIVE: To investigate the clinical relevant factors causing laryngeal cancer postoperative recurrence and analysis the reasons of treatment failure. METHODS: Totally, 574 cases of laryngeal cancer from 1958 to 1999 treated in ENT department of PLA general hospital were retrospectively reviewed. All the patients were observed for 3 years postoperatively, and finally 56 cases of the recurrence were diagnosed. We analyzed the factors possibly causing the recurrence including age, sex, history, time in hospital, smoke, grade of pathology, combination, tumor stage, transfusion, radiotherapy, operation manners and cervical lymph metastasis node grade. RESULTS: By statistic analysis, tumor grade (P = 0.0002), operation (P = 0.0017), transfusion (P = 0.0001), radiotherapy (P = 0.0001) and cervical lymphatic metastasis node grade(P = 0.0228) have significant meaning with laryngeal cancer postoperative recurrence. CONCLUSION: Tumor grade, transfusion, radiotherapy, operation and cervical lymph metastasis node grade are main factors which may cause neoplasm postoperative recurring. PMID- 14743645 TI - [Preliminary study on the correlation factors of cervical lymphatic metastasis of glottic carcinoma]. AB - OBJECTIVE: To investigate the incidence of occult nodal in glottic carcinoma and access the correlation between the cervical lymph nodes metastases and originals of tumor. METHODS: A retrospective review was made on 452 patients from January 1983 to May 1998. 413 patients were male and 39 were female. The age ranged from 42 to 79 years with a mean of 59.3 years. RESULTS: The incidence of occult nodal in glottic carcinoma was low (3.54%), and 0.29% in early stage (T1-T2), 13.39% in late stage (T3-T4), respectively. The number of metastasis lymph nodes was 7 in level II (43.75%), 7 in level III (43.75%) and 2 in level IV (12.5%). CONCLUSION: According to the multivariate analysis, none of the factors such as histopathologic differentiation and invasion of peripheral tissue significantly affect on cervical metastases. The incidence of cervical lymph nodes metastases in glottic carcinoma is low. Selective neck resection should not be undertaken even for the advanced stage that don't exist metastases. PMID- 14743646 TI - Pediatric neurology, Part II. Preface. PMID- 14743647 TI - Childhood autoimmune neurologic diseases of the central nervous system. AB - An autoimmune mechanism for ADEM and MS can be supported by the similar patterns of pathologic changes seen in both diseases with the animal model EAE induced by inoculating animals with nervous tissue and the occurrence of ADEM in patients exposed to nervous tissue during vaccination. Whereas there are no universally agreed-upon criteria for the diagnosis of ADEM, a combination of prodromal illness or preceding vaccination, MRI signs of demyelination, and an acute presentation of neurologic symptoms are the triad most commonly looked for in making the diagnosis of ADEM. An ever-increasing number of infections and vaccinations (nonspecific URIs being most common) has been associated with ADEM. Fever and encephalopathy are seen frequently at presentation. Seizures also are common, as are cranial nerve abnormalities and motor symptoms. A mild pleocytosis or protein elevation is found in the majority of patients with ADEM. Intrathecal IgG synthesis and oligoclonal bands are relatively infrequent but should not be considered inconsistent with the diagnosis of ADEM. White matter changes on T2 in a bilateral although asymmetric distribution with relative sparing of the periventricular region with or without deep gray matter involvement is consistent and to some a requirement for the diagnosis. Low-dose steroids have no beneficial effect in the treatment of ADEM and may be contraindicated. High-dose steroids may have a beneficial effect, particularly in more prolonged illnesses, although the evidence is primarily anecdotal. If steroids are used to improve morbidity, 30 mg/kg/d of methylprednisolone for three to five days is the dose with a six week taper to reduce the risk of recurrence. The prodromal infection may be a major factor in the ultimate mortality and morbidity of the disease. The current mortality of ADEM is quite low. Whether or not this is an effect of different triggering agents or changes in medical care cannot be determined. In larger series of patients with ADEM, 10% to 20% of children experience some sort of recurrence with the majority occurring in the initial one to two months after the first event. This is sometimes associated with steroid withdrawal. A second group of children have a late second recurrence that clinically may not be MS but a recurrence of ADEM, although longer follow-up may change that assessment. Two months should be allowed before a second relapse is considered a manifestation of MS, whereas a second attack also may occur years after an initial attack of ADEM and still be consistent with ADEM recurrence. MS does occur during childhood, with the youngest children at the least risk, and risk increasing with age. The criteria of Poser et al can be used to diagnose MS in childhood [40]. The presentation of MS in childhood is most often sensory, motor, and brainstem signs and symptoms. A relapsing-remitting course is most common with a first relapse occurring in the year after presentation. MRI findings in MS typically show periventricular changes. Oligoclonal bands and CSF IgG synthesis are found in the majority. Treatments of childhood MS have not been studied adequately, but, when treatments studied in adults are used in children, they are well tolerated. Efficacy has not been shown. The long-term outcome of MS in childhood can be either severe or benign with no clear consensus that childhood MS is either a less or more severe disease than the adult form. ATM and ON treatments and outcomes are particularly difficult to evaluate because of the heterogeneity of populations included in case series and the small numbers reported. Steroids are used with anecdotal reports of their superiority to nontreatment. Outcome in ATM often can be poor, whereas in ON it rarely is. A multinational collaborative effort to study and collect the large numbers necessary to address the important questions in these childhood autoimmune disorders would be of great benefit and the only way likely to demonstrate good evidenced-based medicine practiced in this field. PMID- 14743648 TI - Channelopathies in pediatric neurology. AB - Although the genetic neurologic channelopathies are uncommon, they serve as models that further understanding of disease mechanisms in paroxysmal disorders. Many other neurologic channelopathies likely will be identified in the future. PMID- 14743649 TI - Congenital myopathies/dystrophies. AB - The congenital myopathies and congenital muscular dystrophies are a group of relatively infrequent neuromuscular disorders. Ultimate understanding of these disorders, however, will undoubtedly shed considerable light on skeletal muscle development and function. Three classical congenital myopathies are central core disease, nemaline myopathy, and centronuclear myopathy. The congenital muscular dystrophies are often distinguished by whether or not they are associated with clinically evident cerebral involvement. PMID- 14743650 TI - Muscular dystrophy overview: genetics and diagnosis. AB - A specific genetic diagnosis can be reached for most children with muscular dystrophy. Advanced diagnostics, including genetic testing and analysis of nonmuscle tissues, such as skin and blood, often allow the diagnosis to be reached using minimally invasive procedures. These diagnostic advances accompany improved understanding of pathophysiology and pave the way for specific and curative treatments. PMID- 14743651 TI - Mitochondrial encephalopathy. AB - Mitochondrial disorders cause a wide spectrum of diseases in children. Their presentation is nonspecific with encephalomyopathy, failure to thrive, seizures, ophthalmoplegia, and sensorineural hearing loss. These disorders are progressive and are aggravated by fever and infections. They can be caused by mutations in nDNA or mtDNA. Diagnosis requires a complex battery of clinical studies coupled with diagnostic findings on muscle biopsy (abnormal structure, histochemistry, or enzyme studies) or DNA testing. Therapy for mitochondrial disorders remains largely ineffective. PMID- 14743652 TI - Germinal matrix-intraventricular hemorrhage in the premature newborn: management and outcome. AB - Germinal matrix-intraventricular hemorrhage (GMH-IVH) in the premature newborn results from rupture of fragile capillaries in the germinal matrix. Its pathogenesis is multifactorial and relates principally to a pressure-passive cerebral circulation, fluctuations in cerebral blood flow, and derangements of coagulation and fragility of the germinal matrix microvasculature. Several interventions have beneficial effects for prevention of GMH-IVH. Outcome after GMH-IVH relates largely to the severity of hemorrhage, the extent of hemorrhagic and ischemic parenchymal involvement, and complications (e.g., posthemorrhagic hydrocephalus). Even in the absence of neuroimaging abnormalities, VLBW infants have a high incidence of academic and behavioral problems which persist into adolescence and early adulthood. PMID- 14743653 TI - Approach to the rehabilitation of spasticity and neuromuscular disorders in children. AB - The management of children and adults with upper motor neuron disorder is complex and multifaceted. This article reviews new information and potential treatment. As part of the upper motor neuron syndrome (UMNS), spasticity may occur in cerebral palsy, congenital brain malformation, head injury, or other etiologies. Within the UMNS the most recognizable clinical concern is the frequent abnormality of tone, which may have a significant functional impact. Tone reduction is not itself a goal, but is performed for the functional benefits it may allow. New approaches to treatment and management of hypertonia recently have become available. There are many other associated features of the UMNS that affect patient functioning. Ones that frequently occur are abnormalities of speech and other areas of oral motor control. A new area of intervention combines the use of botulinum toxin and ultrasonography to address the common problem of slalorrhea, which is a potential medical issue and a substantial social barrier in affected patients. This article also reviews new information and potential treatment for neuromuscular disorders. PMID- 14743654 TI - Pediatric hydrocephalus: current management. AB - Hydrocephalus is a common problem in pediatric neurology and neurosurgery. The key to the diagnosis of hydrocephalus is the clinical or radiographic progression over time. Most children who have hydrocephalus require ventriculoperitoneal shunts, but the complications of shunting remain distressingly common. Using current endoscopic techniques, a small proportion of patients can be treated without ventriculoperitoneal shunts. The population of pediatric patients who have had hydrocephalic shunts is maturing; these young adults now face issues related to independent living, pregnancy, and long-term care of their hydrocephalus. PMID- 14743655 TI - Pediatric brain tumors. AB - Brain tumors in children vary with regard to histology, location, gender distribution, and age of onset. The past several decades have witnessed substantial improvements in the diagnosis and treatment. As a result, the number of long-term survivors also has increased continuously. Recent advances in neuroimaging facilitate tumor localization and mapping of brain function by noninvasive techniques and are becoming important in preoperative assessment for brain tumors. Surgery remains the mainstay of treatment of many tumor types. A role for chemotherapy is emerging, particularly for nonresectable tumors and in infants, for whom the adverse effects of radiation therapy are severe. Despite the improved treatments and prognostic data, however, many long-term survivors experience significant neurocognitive and developmental deficits. PMID- 14743656 TI - Autism. AB - Autism is a neurologic disorder with impairments in language, social communication, and behavior, which may improve over time, but which persist throughout the lifetime. The evaluation of autism requires a separation of clinical and research objectives and is done best in close cooperation with professionals in the fields of communication, education, and psychology. There are no biologic markers of autism. Regression in language and social communication is present in approximately 30% of children with autism and is most likely to occur between 18 and 24 months of age. Early deficits in social communication can be identified by the assessment of joint attention, affective reciprocity, and metacognition. Current evidence suggests that deficits in social cognition and communication in autism may be related to dysfunction in the amygdala, hippocampus, and related limbic and cortical structures. Other neuroanatomic structures, such as the cerebellum, also may form part of a distributed neuronal network responsible for social cognition and communication. Genetics play a major role in autism, but what is inherited and how broad the inheritable phenotype is remain unclear. At a neurochemical level, the principal neurotransmitter implicated in autism is serotonin. Seizures and epileptiform discharges are common in autism and are linked to cognitive dysfunction. The role of medication is to target specific symptoms and open windows of opportunity that allow implementation of a multimodal individualized educational plan. PMID- 14743657 TI - Attention-deficit/hyperactivity disorder in children and adolescents. AB - ADHD is a common developmental disorder frequently associated with additional neurologic and psychiatric disorders. Stimulant medication is an effective first line therapy for most children with ADHD. Current research is defining the genetic and neurobiologic basis for this disorder. PMID- 14743658 TI - Learning disorders. AB - Three broad areas of LDs are recognized in the DSM-IV (RD, mathematics disorder, and disorder of written expression), and many atypical LDs (such as NVLD) also may be found in children with academic underachievement. These disorders are defined by a significant discrepancy between a child's intellectual (learning) ability and specific area of academic achievement, based on individual psychologic testing. Interventions for these disorders begin with careful evaluation and testing, followed by meetings at the school and development of an IEP. The recommendations of the IEP are performed in the school setting, whereas additional interventions may be sought at private learning clinics. Specific treatments for RD have been well defined, whereas those for other LDs must be tailored to match the strengths and weaknesses of the child. Although it is rare for individuals to completely outgrow the academic weaknesses characteristic of an LD, their performance in the area of weakness often can be improved markedly with appropriate interventions. PMID- 14743659 TI - Depression in children and adolescents. AB - Depression is a common problem in children and adolescents. The disorder may be overlooked because of the prominent irritability seen in children with depression and because of the perception that moodiness is a normal phase of childhood. Depression frequently is associated with other psychiatric problems and neurologic disorders. Therapy consists of psychotherapy and medication, with SSRIs the first choice for pharmacotherapy. PMID- 14743660 TI - Sleep disorders in childhood. AB - Childhood sleep-wake disorders are common and associated with significant impairment of quality of life. The recent discovery of hypocretin deficiency as the pathophysiologic basis for narcolepsy-cataplexy is likely to spur the development of hypocretin analogs for definitive treatment [82,83]. The link between disrupted sleep and daytime learning and childhood inattention is a challenging area of mind-brain interaction in which further progress is likely, once methodologic issues have been sorted out. The recent recognition of genetic influences in the control of circadian rhythms also may spur the development of specific therapies for circadian rhythm disorders. In many ways, sleep medicine is benefiting from recent progress in the basic neurosciences, genetics, and technology. PMID- 14743661 TI - Tuberous sclerosis complex and neurofibromatosis type 1: the two most common neurocutaneous diseases. AB - TSC and NF1 are the most common of the neurocutaneous diseases, and both are autosomal dominant with a high spontaneous mutation rate. For diagnosis, two features are necessary for each disease. Skin findings for each are especially helpful for diagnosis, as is neuroimaging in TSC. For NF1, neuroimaging is not yet reliable for diagnosis. In children, brain symptoms cause most of the morbidity in TSC, and nerve sheath and nervous system tumors as well as learning disabilities cause major morbidity in NF1. Renal disease becomes a serious problem for adults with TSC. The TSC1, TSC2, and NF1 genes function as tumor suppressor genes and have other functions that are being investigated. Blood tests for diagnosis have a high false-negative rate. Therapies for TSC and for NF1 are both medical and surgical. PMID- 14743662 TI - Immunohistochemical changes in kidney glomerular and tubular proteins caused by rattlesnake (Crotalus vegrandis) venom. AB - Renal damage is an important cause of death in patients who have survived the early effects of severe crotalid envenomation. Extracellular matrix of renal tissue is altered by Crotalus toxin activities. The aim of this study was to describe how cytoskeletal proteins and basal membrane components undergo substantial alterations under the action of Crotalus vegrandis crude venom and its hemorrhagic fraction (Uracoina-1) in mice. To detect the proteins in question, the immunoperoxidase method with monoclonal and polyclonal antibodies was used. Cell types within renal lesions were characterized by phenotypic identification, by means of immunohistologic analysis of marker proteins using different primary antibodies against mesangial cells, endothelial cells, cytoskeletal proteins (intermediate filament), extracellular matrix and basal membranes. Samples for morphological study by standard procedures (biotin streptavidin-peroxidase technique) using light microscopy were processed. Positive and negative controls for each antigen tested in the staining assay were included. After crude venom and hemorrhagic fraction inoculation of mice, the disappearance of cytoskeletal vimentin and desmin and collagen proteins in the kidney was observed. In extracellular matrix and basal membranes, collagen type IV from envenomed animals tends to disappear from 24 h to 120 h after venom injection. PMID- 14743664 TI - Electrophoretic protein patterns and numerical analysis of Candida albicans from the oral cavities of healthy children. AB - The aim of this research was to evaluate the protein polymorphism degree among seventy-five C. albicans strains from healthy children oral cavities of five socioeconomic categories from eight schools (private and public) in Piracicaba city, Sao Paulo State, in order to identify C. albicans subspecies and their similarities in infantile population groups and to establish their possible dissemination route. Cell cultures were grown in YEPD medium, collected by centrifugation, and washed with cold saline solution. The whole-cell proteins were extracted by cell disruption, using glass beads and submitted to SDS-PAGE technique. After electrophoresis, the protein bands were stained with Coomassie blue and analyzed by statistics package NTSYS-pc version 1.70 software. Similarity matrix and dendrogram were generated by using the Dice similarity coefficient and UPGMA algorithm, respectively, which made it possible to evaluate the similarity or intra-specific polymorphism degrees, based on whole-cell protein fingerprinting of C. albicans oral isolates. A total of 13 major phenons (clusters) were analyzed, according to their homogeneous (socioeconomic category and/or same school) and heterogeneous (distinct socioeconomic categories and/or schools) characteristics. Regarding to the social epidemiological aspect, the cluster composition showed higher similarities (0.788 < SD < or = 1.0) among C. albicans strains isolated from healthy children independent of their socioeconomic bases (high, medium, or low). Isolates of high similarity were not found in oral cavities from healthy children of social stratum A and D, B and D, or C and E. This may be explained by an absence of a dissemination route among these children. Geographically, some healthy children among identical and different schools (private and public) also are carriers of similar strains but such similarity was not found among other isolates from children from certain schools. These data may reflect a restricted dissemination route of these microorganisms in some groups of healthy scholars, which may be dependent of either socioeconomic categories or geographic site of each child. In contrast to the higher similarity, the lower similarity or higher polymorphism degree (0.499 < or = SD < 0.788) of protein profiles was shown in 23 (30.6%) C. albicans oral isolates. Considering the social epidemiological aspect, 42.1%, 41.7%, 26.6%, 23.5%, and 16.7% were isolates from children concerning to socioeconomic categories A, D, C, B, and E, respectively, and geographically, 63.6%, 50%, 33.3%, 33.3%, 30%, 25%, and 14.3% were isolates from children from schools LAE (Liceu Colegio Albert Einstein), MA (E.E.P.S.G. "Prof. Elias de Melo Ayres"), CS (E.E.P.G. "Prof. Carlos Sodero"), AV (Alphaville), HF (E.E.P.S.G. "Honorato Faustino), FMC (E.E.P.G. "Prof. Francisco Mariano da Costa"), and MEP (E.E.P.S.G. "Prof. Manasses Ephraim Pereira), respectively. Such results suggest a higher protein polymorphism degree among some strains isolated from healthy children independent of their socioeconomic strata or geographic sites. Complementary studies, involving healthy students and their families, teachers, servants, hygiene and nutritional habits must be done in order to establish the sources of such colonization patterns in population groups of healthy children. The whole cell protein profile obtained by SDS-PAGE associated with computer-assisted numerical analysis may provide additional criteria for the taxonomic and epidemiological studies of C. albicans. PMID- 14743663 TI - Human leptospirosis: a twenty-nine-year serological study in Sao Paulo, Brazil. AB - A retrospective study of 9,335 cases of human leptospirosis in the state of Sao Paulo, Brazil, diagnosed between 1969 and 1997 showed that the disease is endemic throughout the state. Middle-aged adults, with a range of 20-39 years, were most frequently infected (32.40%). The mean annual incidence was 0.53 per 100,000 population and the disease was more frequent in males (87.0%). Cases occurred mainly in January to April each year. A peak was observed in 1991 and 1996 which rainfall average was 159.9 and 160.3, respectively. These data emphasize the potential public health importance of leptospirosis in the state of Sao Paulo, Brazil. PMID- 14743665 TI - Dermatophyte agents in the city of Sao Paulo, from 1992 to 2002. AB - Dermatophytosis are superficial mycoses caused by fungi that can invade stratum corneum and keratinized tissues. In order to study the frequency of dermatophytes species and the clinical manifestations caused by these fungi, in Sao Paulo, SP, Brazil, the authors analyzed cultures isolated at the Mycology Laboratory from a selected population (15,300 out-patients of the Hospital das Clinicas, Department of Dermatology, Faculty of Medicine of University of Sao Paulo) from January 1992 to June 2002. The most prevalent dermatophyte was Trichophyton rubrum (48.7%), followed by Microsporum canis (20.9%), Trichophyton tonsurans (13.8%), Trichophyton mentagrophytes (9.7%), Epidermophyton floccosum (4.1%), and Microsporum gypseum (2.5%). These agents determined more than one clinical manifestation, i.e., tinea corporis (31.5%), tinea capitis (27.5%), tinea unguium (14.8%), tinea cruris (13.9%), tinea pedis (9.9%), and tinea manuum (1.9%). Clinical variants of dermatophytosis and their relationship to the etiologic agents were studied and the results were compared to those obtained in previous studies in other regions of Brazil and in other countries. PMID- 14743666 TI - Heterologous antigen extract in ELISA for the detection of human IgE anti Strongyloides stercoralis. AB - Strongyloides ratti larval extract was used for the standardization of ELISA to detect genus-specific IgE in human strongyloidiasis. Forty serum samples from monoinfected patients shedding S. stercoralis larvae (Group I), 40 from patients with other intestinal parasites (Group II), and 40 from copronegative healthy subjects (Group III) were analyzed. Genus-specific IgE levels (ELISA Index: EI) were significantly higher in the group I (EI = 1.43) than groups II (EI = 0.70) and III (EI = 0.71), showing positivity rates of 55%, 2.5% and 0%, respectively. Similarly, sera from copropositive patients had significantly higher levels of total IgE (866 IU/mL) as compared to those from group II (302 IU/mL) and III (143 IU/mL). A significant positive correlation was found between levels of Strongyloides specific-IgE and total IgE in sera from patients with strongyloidiasis. In conclusion, S. ratti heterologous extract showed to be a useful tool for detecting genus-specific IgE by ELISA, contributing for a better characterization of the immune response profile in human strongyloidiasis. PMID- 14743667 TI - Chagas disease in a community in southeast Brazil. I. A serologic follow-up study on a vector controlled area. AB - The prevalence of Trypanosoma cruzi infection was evaluated in Berilo, Minas Gerais (MG), Brazil, from January to July 1997. A serological survey using the indirect immunofluorescence test (IFT) in dried blood collected on filter-paper was performed in a sample of 2,261 individuals. The overall prevalence rate of T. cruzi infection was 18%, and reached 50% in individuals older than 30 years from rural areas. The percentage of seropositivity was 0.17% among individuals younger than 10 years old, suggesting that vectorial transmission is controlled in the area. A decrease in prevalence rates among people born after 1960 and 1970 was observed and this appears to be correlated with the beginning of control programs. A reduction in T. cruzi infection rates was observed when comparing our results with the rates estimated in a serologic study carried out in Berilo in 1983. PMID- 14743668 TI - Pulmonary tuberculosis: relationship between sputum bacilloscopy and radiological lesions. AB - OBJECTIVES: To determine the frequency of radiological manifestations of chest tuberculosis among the tuberculosis outpatients at the Santa Casa de Misericordia de Sao Paulo Hospital, and to correlate these radiological findings with the sputum bacilloscopy. SAMPLE AND METHODS: A review was made of the medical record cards and chest X-rays of all patients attended between January 1996 and December 1998. Patients with a diagnosis of tuberculosis who presented intrathoracic manifestations of the disease and negative anti-HIV serology were selected. RESULTS: The selection included 153 patients, with an average age of 37.5 years, who were predominantly male (60.8%) and white (56.9%). Pulmonary lesions were present in 121 (79.9%) and extrapulmonary lesions in 32 (20.1%). Parenchymal infiltrate lesions appeared in 56 patients (36.6%), cavity lesions in 55 (36.0%), pleural effusion in 28 (18.3%), isolated nodules in 6 (3.9%), mediastinal enlargement in 4 (2.6%) and miliary pattern in 4 (2.6%). Cavities were present in 45.5% of the patients with pulmonary lesions, generally in association with the parenchymal-infiltrate lesions. Parenchymal infiltrate was present in 86.8% of the patients with pulmonary lesions. There was significant presence of alcohol acid resistant bacillus in the sputum of patients with cavities (76.4%), in comparison with those without cavities (50%) (p = 0.003). CONCLUSIONS: Parenchymal-infiltrate lesions are the most frequent radiological manifestation of pulmonary tuberculosis, and they are generally associated with cavities. There is a relationship between the presence of acid fast bacilli in sputum and pulmonary cavity lesions. PMID- 14743669 TI - Evidence of active herpesvirus 6 (variant-A) infection in patients with lymphadenopathy in Belem, Para, Brazil. AB - A total of 323 patients with lymphadenopathy were selected in Belem, Brazil, between January 1996 and December 2001, and screened for the presence of human herpesvirus-6 (HHV-6) IgM- and--IgG antibodies by enzyme-linked immunosorbent assay (ELISA). When seroprevalence is analyzed by gender, similar rates are found for female (60.6%) and male (55.7%) individuals. Seventy-seven (23.8%) patients were HHV-6-IgM-and--IgG-positive (IgM+ subgroup), with positivity rates of 29.7% and 17.7% (p = 0.0007) for female- and male individuals, respectively. Sera from a subgroup (n = 120) of these subjects, with high HHV-6 antibody levels (either IgM+ or IgG+ reactivities), were subsequently processed for the presence of HHV-6 DNA by polymerase chain reaction (PCR)/nested PCR. Active infections (IgM+ and/or IgG+ high levels specific antibodies plus detection of viral DNA) were diagnosed in 20/77 (20.0%) and 8/43 (18.6%); subgroup of the 120 individuals suspected of having HHV-6 suggestive recent infection. All (n = 28) cases of active infection were found to be associated with HHV-6 variant-A (HHV-6A), as detectable by PCR/nested PCR, using variant-specific primer that amplify regions of 195 base pairs (bp) (HHV-6A) and 423 bp (HHV-6B). Rates of HHV-6 DNA detection between female and male patients were similar (p > 0.05) in the IgM+ and IgG+ groups: 20.4% versus 35.7% and 25.0% versus 13.0%, respectively. HHV-6 DNA was detected across < or = 5 through 41-50-year age-groups for patients whose serum samples were IgM+, with rates ranging from 7.7% (female subjects aged < or = 5 years) to 80.0% (male, 11-20 years). Among patients whose serological status was IgG+, HHV 6 DNA was detected in < or = 5, 6-10, 21-30 and > 50 age-groups at rates that ranged from 15.4% (male, < or = years of age) to 100.0% (female aged 11-20 years). Swelling cervical lymph nodes were the most common sign, accounting for 9 (32.0%) cases in each gender group. Among patients (n = 28) with active infection by HHV-6A variant, duration of symptoms lasted 1-5 days in 35.7% of subjects, whereas in 64.3% of them the disease lasted 6-20 days. Our data suggest that it is worth seeking for HHV-6 infection whenever a patient (infant or adult) presents with lymphadenopathy as a prominent symptom in the course of an acute febrile illness. PMID- 14743670 TI - Evaluation of the formalin-tween concentration technique for parasitic detection. AB - The formalin-Tween sedimentation method was compared with the formalin-ether sedimentation for parasitic detection. Of a total 297 fecal specimens examined, 72.1% were positive. The formalin-tween technique was effective for ascertaining helminths, particularly Ascaris lumbricoides, Trichuris trichiura and hookworm eggs; however it has less capability for protozoa detection. This method is simple, inexpensive, less time consuming and highly sensitive when detecting the parasitic infection, particularly when focusing on helminth eggs. PMID- 14743671 TI - Human toxocariasis: incidence among residents in the outskirts of Campinas, State of Sao Paulo, Brazil. AB - With the aim of estimating the incidence of infection by Toxocara among residents in the outskirts of Campinas (State of Sao Paulo, Brazil) two serological surveys, using ELISA anti-Toxocara tests, were performed in January 1999 and January 2000, involving, respectively, 138 and 115 individuals, 75 of which examined in both occasions. Among this group 67 individuals did not show the presence of anti-Toxocara antibodies in 1999, and 12 presented seroconversion in the second survey, revealing an annual incidence rate of 17.9%. PMID- 14743673 TI - All in the family. PMID- 14743672 TI - Diagnosis of abdominal angiostrongyliasis by PCR from sera of patients. AB - Abdominal angiostrongyliasis is a zoonotic infection caused by an intra-vascular nematode parasitic of wild rodents, Angiostrongylus costaricensis. No parasitological diagnosis is currently available and immunodiagnosis presents several drawbacks. Primers constructed based on a congeneric species, A. cantonensis, were able to amplify a 232 bp fragment from serum samples of 3 patients with histopathological diagnosis. Extraction was better performed with DNAzol and the specificity of the primers was confirmed by Southern blot. This disease has been diagnosed with frequency in south of Brazil, thus, this method appears like the important and unpublished alternative to improve diagnostic of disease. PMID- 14743674 TI - Winter warming. PMID- 14743675 TI - Lessening the burden. PMID- 14743676 TI - Under the surface. PMID- 14743677 TI - Prescribing independence. PMID- 14743678 TI - Divided attention. PMID- 14743679 TI - Finding context. PMID- 14743680 TI - The co-location equation. PMID- 14743681 TI - In with the new. Landmark Medicare legislation provides relief for rehab providers. PMID- 14743682 TI - [Microsatellite instability and loss of heterozygosity in neoplastic and preneoplastic gastric lesions]. AB - BACKGROUND: Gastric cancer is the leading cause of cancer deaths in the general population in Chile, with mortality rates as high as 33.7 per 105 in males in the IX region. A chain of genetic and morphological events precedes the intestinal type of gastric carcinoma. One of them is the called multifocal atrophic gastritis often associated with intestinal metaplasia. AIM: To study the frequency of microsatellite instability (MSI) and loss of heterocigozity (LOH) in neoplastic and preneoplastic lesions of gastric carcinoma, especially intestinal metaplasia. MATERIAL AND METHODS: Ninety four gastric cancer biopsies were studied using laser capture microdissection, to obtain well defined cell populations from paraffin-embedded tissues: lymphocytes (control DNA), intestinal metaplasia and gastric cancer areas. Primer flanking microsatellite 15 highly polymorphic regions were used to study MSI and LOH. Radioactive PCR products were electrophoresed and exposed for autoradiography. RESULTS: LOH was observed in 83% of gastric carcinomas and in 54% areas containing intestinal metaplasia. The most commonly altered regions were the CA repeat associated with the p53 gene and the 3p21 region. High grade MSI was observed in 11.7% of gastric cancer preparations and 17% of intestinal metaplasia associated to cancer with MSI-H phenotype. CONCLUSIONS: MSI and LOH were frequently observed in intestinal metaplasia glands in patients with gastric carcinoma. The frequency of MSI-H phenotype in gastric patients was slightly lower than the one described in sporadic colorectal cancer not associated to HNPCC. The high incidence of genetic lesions in intestinal metaplasia area, support the idea that intestinal metaplasia is a genetically highly unstable cell population. PMID- 14743683 TI - [Clinical and electrophysiological characteristics of patients with atrioventricular nodal reentry tachycardia who underwent slow pathway ablation]. AB - BACKGROUND: AV nodal reentry tachycardia (AVNRT) is the most common cause of paroxysmal supraventricular tachycardia. Radiofrequency ablation is today the treatment of choice. AIM: To report our experience in patients who underwent slow pathway ablation. PATIENTS AND METHODS: Fifty six consecutive patients (68% female, mean age 43 years old) that underwent slow pathway ablation are reported. RESULTS: Sixty four percent of patients had failed drug therapy. During electrophysiological study, AVNRT was induced in 55 patients. Isoproterenol was required for induction in 36%. Programmed atrial stimulation revealed dual AV nodal pathway in only 64% of the patients; 29% had AVNRT with single nodal curve and 7% only prolongation of AH interval. The slow pathway was ablated in 55 patients. One patient refused ablation because of risk of AV block. All patients had immediate success post ablation. Sixty four percent of patients persisted with partial evidence of dual curve manifested by sudden AH prolongation and single echoes. CONCLUSIONS: Isoproterenol is essential for ruling out AVNRT, since 29% of the patients had baseline single nodal curve and in only 64% was tachycardia induced without isoproterenol. Persistence of residual dual physiology does not rule out the success of ablation. PMID- 14743685 TI - [Sexual function in postmenopausal women using hormone replacement therapy]. AB - BACKGROUND: The loss of sexuality observed in the climacteric period is difficult to evaluate. An important advance has been the development of the Female Sexual Function Index (FSFI), a test based on the norms of the International Consensus Development Conference on Sexual Female Dysfunctions. AIM: To study the effects of hormone replacement therapy (HRT) on sexuality, applying the FSFI. MATERIAL AND METHODS: The FSFI was applied to 300 healthy women between 45-64 years, sexually active, beneficiaries of the Southern Metropolitan Health Service. RESULTS: The mean age of the sample studied was 51 +/- 5 years, 27% were HRT users, 21% had had an hysterectomy and 98% had a stable couple. The total score of the FSFI decreased from 27.3 +/- 5.8 in women between 45 and 49 years of age to 19.3 +/- 7.0 in women between 60 and 64 years (p < 0.01). A significantly better sexuality was observed in HRT users, with FSFI scores of 28.1 +/- 5.5 and 24.6 +/- 6.8 in HRT users and non users, respectively (p < 0.01). Women on HRT obtained a higher score in all of the test domains, especially in lubrication, orgasm and sexual satisfaction. CONCLUSIONS: Female sexuality decreases with aging. HRT users have a better sexual function than non users. PMID- 14743684 TI - [Validation of the chronic respiratory questionnaire in Chilean patients with chronic airflow limitation]. AB - BACKGROUND: Chronic respiratory diseases are an important problem in Chile, causing disability and worsening the quality of life. AIM: To validate the Spanish version of the Chronic Respiratory Questionnaire (CRQ) to be used in Chilean population. MATERIAL AND METHODS: The CRQ was applied to 38 patients with chronic respiratory diseases. At the same time the functional and physiologic status of patients (the 6 Minute Walking Test and spirometry) was assessed in an independent and blinded fashion. RESULTS: CRQ scores were associated with 6 Minute Walking Test and forced expiratory volume results (p < 0.01 adjusted by age, sex and smoking). The Internal Consistency analysis showed a Cronbach alpha between 0.60 and 0.89. CONCLUSIONS: Considering the construct validation as an on going process, the present results give additional evidence supporting the use of the CRQ in Chilean population to evaluate the health-related quality of life of patients with chronic respiratory diseases. PMID- 14743686 TI - [Evaluation of mental and communication functions in mapuche and non mapuche elderly subjects in rural communities in Southern Chile]. AB - BACKGROUND: The main predictors of functional impairment in the elderly are alterations in mental or communication functions. AIM: To study mental and communication functions in rural elderly subjects of Mapuche and non Mapuche origin. PATIENTS AND METHODS: Elderly subjects coming from a rural Mapuche community and a non Mapuche community were studied. Subjects were interviewed at their homes. The communication and mental function assessments of the Functional Autonomy Measurement System were applied. RESULTS: Fifty one Mapuche and 49 non Mapuche subjects with a mean age of 71 +/- 7 and 74 +/- 8 years respectively, were studied. Fifty four percent were female and 31% were illiterate. Twenty six percent had impairment in mental functions. The item with the highest difficulty was memory. The visual function was the most severely impaired among communication items. Mapuche elderly subjects had significantly higher degrees of impairment in mental and communication functions. CONCLUSIONS: There is a higher degree of mental and communication impairment among rural Mapuche elderly subjects than in their non Mapuche counterparts. PMID- 14743687 TI - [10-year experience in flexible bronchoscopy in pediatric patients]. AB - BACKGROUND: Approximately a decade ago, pediatric Flexible Bronchoscopy (FB) was introduced in Chile, after being used for several years in adults. AIM: To describe our clinical experience in FB in a ten years period. PATIENTS AND METHODS: Records of procedures done between January 1993 and September 2002 at the Pediatric Service of the Catholic University Hospital, were retrospectively reviewed. We evaluated the clinical indications for the procedures in relation to patient's age and the correlation between indications and FB findings. RESULTS: A total of 700 procedures were performed during the period, 59% in men and 53% in patients younger than 1 year. Seventy seven percent of procedures were done in an examination room, using a nasal approach. The main indication was visualization of the airway (49%). The most common clinical diagnosis, in descending order were: atelectasis, stridor and etiologic study of pneumonia by bronchoalveolar lavage (BAL). In children younger than 6 months the most common clinical diagnosis was stridor, followed by atelectasis. The main diagnosis in the whole sample, reached by FB was atelectasis secondary to mucous plug. In children younger than 6 months, the main diagnosis was laryngomalacia. A positive microbial culture was obtained in 43% of patients in whom BAL was done. Complications were uncommon (5%) and mostly mild. In 2.3% of cases, these were severe, such as bronchospasm and need for mechanical ventilation. Severe complications were observed in patients younger than 3 months with severe stridor or in children with cancer, who required FB and BAL. CONCLUSIONS: Flexible bronchoscopy is a safe and useful procedure in pediatric patients. PMID- 14743688 TI - [Study of collagen and elastic fibers of connective tissue in patients with and without primary inguinal hernia]. AB - BACKGROUND: There are few studies looking for collagen matrix defects in patients with inguinal bernia. AIM: To study the skin connective tissue in patients with and without inguinal bernia. PATIENTS AND METHODS: Skin from the surgical wound was obtained from 23 patients with and 23 patients without inguinal bernia. The samples were processed for conventional light microscopy. Collagen fibers were stained with Van Giesson and elastic fibers with Weigert stain. RESULTS: Patients without hernia had compact collagen tracts homogeneously distributed towards the deep dermis. In contrast, patients with hernia had zones in the dermis with thinner and disaggregated collagen tracts. Connective tissue had a lax aspect in these patients. Collagen fiber density was 52% lower in patients with hernia, compared to subjects without hernia. No differences in elastic fiber density or distribution was observed between groups. CONCLUSIONS: Patients with inguinal bernia have alterations in skin collagen fiber quality and density. PMID- 14743689 TI - [Phenylketonuria diagnosed during the neonatal period and breast feeding]. AB - BACKGROUND: Phenylketonuria (PKU) is due to of a defect in the phenylalanine hydroxylase gene (12q22-24.1) leading to hyperphenylalaninemia. Treatment consists in a low phenylalanine (Phe) diet. AIM: To evaluate the evolution of early diagnosed PKU children, receiving direct breast feeding, and a special formula without Phe, during their first six months of life. PATIENTS AND METHODS: Nineteen PKU children diagnosed in the neonatal period (19.29 +/- 13.8 days of age), treated with breast feeding and formula without Phe since diagnosis, were studied. Intake of calories, proteins and dietary Phe were quantified. Blood Phe, nutritional status and psychomotor development were also measured. RESULTS: The diet that these children received during the 6 months period of study, had a mean of 127 +/- 19.9 Kcal/kg/day, 1.95 +/- 0.3 g protein/kg/day and 35.3 +/- 9.5 mg Phe/kg/day. Fifteen children maintained the blood level of Phe under 8 mg/dl, considered an excellent metabolic control. Only 4 cases had intermittently high levels, between 10-12 mg/dl. At 6 months of age, 74% of the children maintained breast feeding as the only source of Phe. Sixty three percent had a normal nutritional status, 5.2% were at nutritional risk and 31.6% were overweight. Eighty one percent had a normal mental development. CONCLUSIONS: The use of direct breast feeding allows a good metabolic control and improves growth and development of early diagnosed PKU children. PMID- 14743690 TI - [Bone mineral density in school age children born preterm ]. AB - BACKGROUND: The age at which children born preterm normalize their bone mineral density, is not well known. AIM: To study if children born preterm have normalized their bone mineral density at age 5 to 7 years. PATIENTS AND METHODS: Twenty six infants born preterm (14 male), were studied at age 5 to 7 years. Birth weight, present weight and height, bone age, calcium and phosphate intake at the first year of life and at the current age were assessed. Bone mineral density was measured by single photon X ray absorptiometry in the dominant forearm. A blood sample was obtained to measure insulin growth factor 1 (IGF-1). As a control group, 105 healthy age-paired infants born at term, were studied. RESULTS: Bone mineral density was significantly lower in infants born preterm than in their term counterparts (0.273 +/- 0.01 g/cm2 and 0.302 +/- 0.01 g/cm2 respectively, p < 0.001). There was a positive correlation between bone mineral density and IGF-1 (r = 0.49, p = 0.01). No correlation with the other measured parameters was observed. CONCLUSIONS: Infants that were born preterm have a lower bone mineral density at 5 to 7 years of age than their term controls. Bone mineral density correlates with IGF-1. PMID- 14743691 TI - [Characterization of species of the gender Malassezia in patients with seborrheic dermatitis and subjects without skin lesions]. AB - BACKGROUND: The yeast Malassezia spp has an established etiological role in pityriasis versicolor, folliculitis, systemic infections and onychomycosis. AIM: To assess the presence of Malassezia spp in patients with seborrheic dermatitis (SD), to find a correlation between Malassezia spp count and the severity of the disease and to compare the prevalence of the different Malassezia species in SD patients and subjects without skin lesions. PATIENTS AND METHODS: Scrapings of the face from 81 patients with SD (69 males) and 79 subjects (54 males) without skin lesions were obtained for a direct microscope examination and yeast culture. RESULTS: The yeast Malassezia was found in 76% of SD patients and in 82% of subjects without skin lesions. There was a positive correlation between the number of yeasts found on direct examination and the clinical severity of lesions in SD patients. Although this correlation was statistically significant (P = 0.046), the degree of association (rho = 0.22) was weak. Fifty Malassezia species were identified. M globosa was found in 67% of SD patients, followed by M furfur and M sympodialis, each present in 16.5% of the SD patients. In subjects without skin lesions, the most prevalent species were M globosa (77%), followed by M sympodialis (12%), M slooffiae (7%) and M furfur (4%). CONCLUSIONS: The presence of the yeast Malassezia is not associated with the presence of skin lesions. PMID- 14743692 TI - [Acute Liver Failure in patient with liver amyloidosis associated to multiple myeloma]. AB - We report a 67 years old woman admitted to the hospital for the study of a cholestatic jaundice and massive hepatomegaly. On admission, the patient did not have liver failure. During hospital stay, the patient experienced a progressive deterioration of liver function and a monoclonal gammopathy was detected. An IgG Kappa myeloma-was diagnosed. A fine needle liver biopsy disclosed the presence of amyloid. The patient developed acute liver failure and died three weeks after admission. PMID- 14743693 TI - [Diabetic mastopathy. Report of a case]. AB - Diabetic mastopathy is a rare condition that occurs in type I diabetic patients and that may mimic a breast cancer due to its intense fibrosis and inflammation. We report a 34 years old female consulting for a firm, painless and mobile retroareolar breast lump measuring 3.5 x 4 cm, that lead to the suspicion of a breast cancer. Mammography, mammary ultrasound and fine needle cytology were unspecific. A core biopsy confirmed the diagnosis of diabetic mastopathy. The patient continues under periodical assessments. PMID- 14743694 TI - [Combined liver-kidney transplantation: report of a case]. AB - End stage renal disease is not an absolute contraindication for liver transplantation (LT) in patients with end stage liver disease. Actuarial patient and graft survival are comparable for children and adults who undergo LT alone and liver-kidney transplantation (LKT). The most common indications for LKT are the primary hyperoxaluria type I (PH1) and the liver and renal polycystic disease. We report a 12 years old boy with congenital hepatic fibrosis with severe portal hypertension, encephalopathy and polycystic kidney disease with end stage renal disease on dialysis that underwent LKT. During the second postoperative week, he had a biopsy-proven acute liver and renal rejection, that had a good response to corticosteroids. Thirty days after surgery, the liver biopsy was without rejection. No other complications were observed. PMID- 14743695 TI - [Antimicrobial resistance of Helicobacter pylori: clinical and molecular aspects]. AB - Helicobacter pylori is a relevant pathogen for gastroduodenal diseases in human beings. Although its eradication often improves gastroduodenal diseases, H pylori is acquiring an elevated rate of resistance to various antimicrobials, such as metronidazole, clarithromycin, tetracycline and amoxicillin. Multi-drug resistance is a major problem to select the appropriate treatment of infectious diseases. To improve our understanding on the complexity of the problem, in this article we review the resistance mechanisms and give an update on H pylori antimicrobial resistance. PMID- 14743696 TI - [Dietary phytoestrogen and its potential benefits in adult human health]. AB - Human diet contains a series of bioactive vegetal compounds that can improve human health. Among these, there has been a special interest for phytoestrogens. This article reviews the evidence about the potential benefits of phytoestrogens for human health. Forty eight manuscripts were selected for their study design and relevance to human health. The cell growth inhibitory effects of phytoestrogens and their implication in breast cancer are reviewed. Also the effects of these compounds on serum lipid levels and the effectiveness of a phytoestrogen derivate, ipriflavone, on the prevention of osteoporosis are analyzed. Although these compounds have a great potential for improving health, there is still not enough evidence to recommend the routine use of phytoestrogens. PMID- 14743697 TI - [Bioethical aspects of health care reform in Chile. II. Discrimination, free election and informed consent]. AB - Bioethical issues emerge each time health care reform projects are discussed. These affect diverse moral values and principles and have an impact on cultural, social and political areas. Thus, they demand more than just organizational, financial or administrative solutions. This review analyses discrimination, free election of professionals and informed consent. All three concepts are alluded in the legislative debate raised upon the actual process for health reform. Having clear ideas about these subjects is crucial to foresee the reactions expected to arise among physicians and the general public, when confronting the proposed changes. PMID- 14743698 TI - [Fifty years of the heart-lung machine. Report on the pioneers and heroes and about the circumstances that led to the great invention, which allowed the treatment, and in many cases, the cure of heart illnesses]. AB - In 1953 DNA was discovered and the Everest was conquered but also a great invention was developed: the heart-lung machine, which allowed the treatment, and in many cases, the cure of most cardiovascular illnesses. In fact, on May 6, 1953 John Gibbon crowned with success the work of his entire life closing for the first time an atrial septal defect in a young woman using a heart-lung machine of his own invention. Before that, surgeons had explored other roads like hypothermia, cooling the patient in a cold water tub and then rapidly performing the surgical correction of a heart malformation. After his first success, the following 4 patients of Gibbon died, which led him to abandon heart surgery and produced a generalized pessimism about extracorporeal circulation. However, a year later Walton Lillehei reverted this situation with the introduction of controlled cross-circulation in which a patient, usually a child, was connected to a "donor", usually his father or mother, whose heart and lung served as a pump and oxigenator, allowing the performance of open heart surgery. Finally, it was Lillehei again who a year later introduced the bubble oxigenator, simple and inexpensive, opening the doors of open heart surgery to all surgeons around the world. For this, and many other reasons, Walton Lillebei is considered by most surgeons as the "Father of Open Heart Surgery". Lillehei visited Chile in 1963 and operated on a patient in the surgical theaters of the Hospital Clinico de la Universidad Catolica and was named an Honorary Member of the School of Medicine of this University. Before that, in 1957 Helmut Jaeger at the Hospital Luis Calvo Mackenna performed the first successful surgical closure of an atrial septal defect with extracorporeal circulation in Chile using a De Wall-Lillebei bubble oxigenator. PMID- 14743699 TI - [Comments on possible consequences of health reform plans in Chile]. PMID- 14743700 TI - [Effects of antagonists of the various subtypes of dopamine receptors on the locomotor activity of C57BL mice induced by psychostimulators]. AB - The involvement of dopamine receptors of various subtypes in the mechanisms of locomotor hyperactivity caused by single administration of psychostimulants d amphetamine and syndnocarb was studied in C57BL mice. Administered in a dose not influencing the spontaneous locomotor activity, SCH23390 and raclopride- selective antagonists of the D1 and D2 subtypes of dopamine receptors, respectively--abolished the hyperactivity induced by d-amphetamine, while not significantly changing the effect of sydnocarb. On the contrary, clozapine- antagonist of the D4 subtype of dopamine receptors--failed to reduce the stimulant action of d-amphetamine, but significantly decreased the effect of sydnocarb under the same experimental conditions. PMID- 14743701 TI - [Effect of polidan on the conditioned memory and structural and metabolic parameters of the rat brain neurons]. AB - New experimental data on the mnemotropic (nootropic) properties of the well-known drug polidan (a DNA derivative) are summarized. The results obtained by histological techniques showed changes in the brain structure, indicating protein synthesis activation under the action of polidan. These results confirm prognostic function of the concept of evolutionary-molecular basis of memory. PMID- 14743702 TI - [Membrane mechanisms of effects of antihypoxic agents bemethyl and almide on neurons of Mollusca]. AB - Membranotropic effects of the antihypoxants bemithyl and almide, structural analogs of thiobenzimidazole, have been studied on the isolated neuronal preparations of Lymaea stagnalis branchycephalic mollusk. Both drugs in a concentration range of 100-1000 microM produced a reversible, dose-dependent nonselective single-phase blocking action upon the ion channels and completely blocked the channels at a concentration of 10 mM. Therefore, bemithyl and almide are active membranotropic compounds capable (in sufficiently high concentrations) of changing the conductivity of slow sodium, calcium, and potassium ion channels in excitable cells. The protective antihypoxant drug reactions on a systemic level of the organism are probably related to the fact that both drugs in small concentrations are capable of hyperpolarizing the cell membrane, activating the ion channel function, and stabilizing the action potential under hypoxia conditions; in greater concentrations, bemithyl and almide are capable of blocking ion currents, thus reducing the excitability of cells and protecting them from overstress. PMID- 14743703 TI - [Anti-amnestic activities of 3-hydroxy-3-pyrroline-2-one derivatives]. AB - The antiamnesic properties of a series of new 3-hydroxy-3-pyrrolin-2-on derivatives structurally analogous to pyrrolidinone nootropes and GABA were studied on a model of amnesia of the passive avoidance conditioned reflex. The screening revealed compounds whose antiamnesic action is comparable with that of piracetam. PMID- 14743704 TI - [Effect of cinnarizine on the brain mitochondrial oxidative system, antioxidant blood activity, and the rat behavior in hypoxia]. AB - The effect of cinnarizine on the functional state of brain mitochondria, the activity of blood antioxidant system, and the behavior of rats was studied under model hypoxic hypoxia conditions. A four-day treatments with cinnarizine (50 mg/kg, twice per day via a gastric tube) prevents the hypoxic brain edema development, restores NAD+ dependent oxidation of a succinate substrate, normalizes emotional-exploratory activity, and causes hyperlocomotion of the experimental animals, while not influencing a high level of activity of the blood antioxidant system. PMID- 14743705 TI - [Experimental characteristics of the neurotropic spectrum of some antiepileptic agents]. AB - Investigation of the psychotropic spectrum of phenobarbital, diphenin, milontin, pufemide, sodium valproate (depakine), and diazeparn showed that, besides antagonism with the corazole and electroshock effects, all these drugs prevented to a considerable degree the kindling seizures in rats on a "tonsil swing" (chronic electroencephalographic partial epilepsy) model. The mechanism of this effect probably involves exciting amino acids. All anticonvulsants except for diazepam also restored memory in experimental animals with retrograde electroshock amnesia. The administration of diazepam, diphenin, and sodium valproate to rats with kindling convulsions led to activation of the spontaneous horizontal motions in the open-field test, which can be explained by the presence of antianxiety component in the drug action. PMID- 14743706 TI - [Effect of amiodarone on the extrasystolic and post-extrasystolic contractions of the isolated myocardium of rats and of patients with chronic ischemic heart disease]. AB - Peculiarities in the extrasystolic and post-extrasystolic contractility of the isolated myocardium of patients suffering from chronic ischemic heart disease (IHD), in comparison to that of intact rats, were studied in the presence of the antiarrhythmic drug amiodarone. The contractility was measured in an isometric mode at an electric stimulation frequency of 0.5 Hz, a temperature of 37 degrees C, and a perfusion rate of 10 ml/min. The extrasystolic action was produced by additional electric pulses generated 25 sec after the base stimulation signal. Amiodarone (1 microgram/ml) reduced the inotropic response of intact rat myocardium to the extrasystolic action, but increased the post-extrasystolic contractility. In the myocardium of IHD patients, the drug also suppressed manifestations of the spontaneous abnormal contractility, while not decreasing the inotropic response to the extrasystolic action. An increase in the post extrasystolic contractility reached on the average 24% (against 6% in the control). The effect of amiodarone on the post-extrasystolic contractility can be related to a change in the functional state of sarcoplasmic reticulum (SR) and in the content of deposited Ca2+ ions. SR is an important factor of intracellular homeostasis of cardiomyocytes during IHD and can probably influence the antiarrhythmic efficacy of amidarone in IHD patients. PMID- 14743707 TI - [Nephroprotective effect of bilobil in experimental acute kidney failure]. AB - Bilobil, a commercial preparation based on Ginkgo biloba extract, improves the functional state of kidney and increases the survival of rats with glycerol induced acute renal insufficiency. One possible mechanism of this effect is related to the ability of bilobil to simulate the renal antioxidant system, thus decreasing the rate of lipid peroxidation processes and reducing the renal damage. The results show good prospects for the use of bilobil in nephrology. PMID- 14743708 TI - [Effect of himantane on the rat embryo development]. AB - Himantane introduced via a gastric tube to pregnant rats in a dose of 10, 30, 50, and 100 mg/kg produced a dose-dependent embryotoxic and teratogenic action. An analysis of the experimental results and published data suggests that the embryotoxicity of himantane can be related to its general toxic action upon the organism of pregnant female rats. PMID- 14743709 TI - [Effects of the novel synthetic fatty acid dinitroglycerol esters on aggregation of the human platelets in vitro]. AB - A series of six new synthetic dinitroglycerol esters of fatty acids on the human platelet aggregation was studied in vitro. Inclusion of the dinitroglycerol moiety into the molecule of arachidonic acid deprived this acid from pro aggregant activity. All six compounds produced moderate (dose-dependent) inhibition of the platelet aggregation process induced by arachidonic acid (1 x 10(-3) M). Platelet aggregation was most significantly affected by dinitroglycerol esters of arachidonic and docosahexaenoic acids. This is probably explained by the influence of these esters on the oxidative metabolism of arachidonic acid to eicosanoids playing the role of proaggregants. In the presence of vessel wall (rat aorta fragments), dinitroglycerol esters of arachidonic and docosahexaenoic acids incubated with platelets (5 min, 37 degrees C) significantly reduced their aggregation induced by arachidonic acid (1 x 10( 3) M) or docosahexaenoic acid (1 x 10(-5) M) under the conditions of endothelial cyclooxygenase suppressed by acetylsalicylic acid (10 mg/ml). The pronounced antiaggregant effect of the synthetic dinitroglycerol esters studied is probably related to their ability to act as NO donors suppressing the activity of thrombocytes (provided that the NO production activity is present in the system). PMID- 14743710 TI - [Combined use of an antihypoxic agent and a B-cell stimulator]. AB - Evaluation of the effect of poviargol on the B cells of mice showed that the drug activity is retained for 21 h upon single injection. An analysis of the refractory period between the B-cell stimulant introduction and the administration of various antihypoxants showed that the most effective treatment is provided by poviargol injection 14 days before and antihypoxant (IEM-1875 preparation) 30 min before the hypoxia model induction. This regime, ensuring the best protection of experimental animals from hypoxia, can be of great importance and value in the prophylaxis of hypoxia in various pathological conditions. PMID- 14743711 TI - [Effects of anxiolytic agents on some immunologic parameters of the stressed rats]. AB - Repeated stress under forced swim test conditions suppresses most of the immunological characteristics in rats. Diazepam and melatonin (both in a dose of 0.1 mg/kg), while not changing significantly the immunological state of intact animals, decrease the stress-induced shifts. The immunotropic effect of melatonin is much more pronounced than that of diazepam. PMID- 14743712 TI - [Anti-inflammatory activity of the novel water soluble gossipol derivative mebavine]. AB - Investigation of the effect of mebavin on the adjuvant arthritis showed that this water-soluble derivative of gossypol possesses an antiinflammatory activity comparable with that of prednisolone. PMID- 14743713 TI - [Characteristics of pyrrolidone pharmacokinetics in rats]. AB - The pharmacokinetics of pyrrolidone in a composition with pyroglutamic acid was studied in white mongrel male rats. The former component exhibits cerebrovascular and neuroprotector activity. Pyrrolidone, detected in the blood plasma and brain for 8 h after peroral and intravenous administration, exhibits a high absolute bioavailability and the ability to penetrate via the blood-brain barrier. PMID- 14743714 TI - [Preclinical study of maxar safety]. AB - The results of preclinical safety evaluation of the new hepatoprotector maxar showed that this drug can be classified as a low-toxicity substance with respect to acute toxicity. No significant functional and structural changes in the systems and organs of experimental animals were observed after a 6-month administration in rats (in a dose of 300, 600, and 1200 mg/kg) and in dogs (500 mg/kg). Maxar exhibited no mutagen and allergen properties, produced no immunotoxicant action, and did not adversely affect the reproduction function. PMID- 14743715 TI - [Neuroprotective effect of cytoflavin in closed craniocerebral trauma]. AB - The efficacy of a complex preparation cytoflavin was studied on rats with a model of closed craniocerebral trauma. Cytoflavin produced normalization of the energy exchange, lipid peroxidation, and antioxidant system functioning in nervous tissues. The treatment significantly reduced the degree of posttraumatic edemation in the brain. The cerebroprotective action of cytoflavin was somewhat superior to the effect of the reference drug solcoseryl. PMID- 14743716 TI - [Effect of quamatel on neutrophil properties, parameters of lipid peroxidation and the antioxidant defense system in blood of patients with duodenal ulcer and pancreatitis]. AB - The properties of neutrophils, the parameters of lipid peroxidation, and the characteristics of antioxidant protection (superoxide dismutase, catalase, glutathione peroxidase, and glutathione reductase activity) were studied in the blood of patients with duodenal ulcer and pancreatitis in the course of quamatel administration. The pattern of changes induced by quamatel shows evidence of the antioxidant activity of the drug. PMID- 14743717 TI - [Peripheral effects of ligands of ORL1 receptors]. AB - It has been discussed literature data on the role for ORL1 (NOR) receptors in the regulation of function of gastrointestinal, respiratory, cardiovascular, immune, endocrine systems. In addition, it has been discussed a possibility of penetration of blood brain barrier for ORL1 receptor ligands and species dependence of NOR-ligands' effects. PMID- 14743718 TI - Can American afford tomorrow's retirees: results from the EBRI-ERF Retirement Security Projection Model. PMID- 14743719 TI - A waste of energy. PMID- 14743720 TI - Not so icy stares. PMID- 14743721 TI - Doping by design. PMID- 14743722 TI - Becoming behemoth. PMID- 14743724 TI - The great migration. PMID- 14743723 TI - LASH out. PMID- 14743725 TI - Micro(mechanical)phones. PMID- 14743726 TI - Working the system. PMID- 14743727 TI - A bounty of science. PMID- 14743728 TI - Talking bacteria. PMID- 14743730 TI - Four keys to cosmology. PMID- 14743729 TI - Insights into shock. PMID- 14743731 TI - The cosmic symphony. PMID- 14743732 TI - Reading the blueprints of creation. PMID- 14743733 TI - From slowdown to speedup. PMID- 14743734 TI - Out of the darkness. PMID- 14743735 TI - Better displays with organic films. PMID- 14743736 TI - The case of the unsolved crime decline. PMID- 14743737 TI - A walk in the woods. PMID- 14743738 TI - [Levels of granzyme B, perforin and Fas ligand antigens and mRNAs in patients following allogeneic hematopoietic stem cell transplantation]. AB - In order to monitor the immunological status of patients after allogeneic hematopoietic stem cell transplantation(HSCT), granzyme B(GrB), perforin(PRF) and Fas ligand(L) antigens and mRNAs were measured by flow cytometry and real time RT PCR, respectively. Cytoplasmic antigens were detected in whole blood after fixation and pretreatment with saponin. Real time PCR was carried out using extracted RNA from buffy coat. We measured these substances in a cytotoxic T cell clone, a natural killer cell line, and peripheral blood collected from 11 patients after HSCT. Although changes in antigen levels were not detected, increased levels of GrB and Fas L mRNAs were quantitatively measured in CTLs and NK cells stimulated by IL-2 combined with IL-12. Increased levels of GrB and/or PRF antigens were detected in four of five patients with chronic GVHD. Increased mRNA levels were also observed in one or more of GrB, PRF or Fas L in four of five patients with cGVHD, although there was a discrepancy between antigen and mRNA positivity. Four of six patients without cGVHD were positive for apoptosis inducing factors, either by antigen detection or RT-PCR. One of these four had relapsing leukemia, and another had herpes zoster infection, while the reasons for positive results in the other two patients are not clear. Although changes in antigen levels did not parallel those in mRNA, measurement of these parameters may assist in predicting GVHD, GVL and infections following HSCT. PMID- 14743739 TI - [Molecular-epidemiological evaluation of Serratia marcescens isolates from patient specimens in one Japanese hospital during the three years from April 1999 to March 2002]. AB - One hundred fifty-six isolates of Serratia marcescens from patient specimens in Showa University Fujigaoka Hospital between April 1999 and March 2002 were investigated in this study. Forty-two isolates with serotype O2, detected mainly from patient respiratory specimens, were susceptible to the antimicrobial agents tested, whereas 30 isolates with serotype O14, detected mainly from patient urine, were resistant. Moreover, 19 isolates with serotype O14 susceptible to imipenem were intermediate or resistant to meropenem, while they did not produce metallo beta-lactamase. Both serotypes were significantly distributed in the ICU and surgical wards, compared with other wards. Ten isolates with O2/B (bacteriocin type) 16 and 14 isolates with O14/B76J, showing identical or closely related clones by pulsed-field gel electrophoresis (PFGE) analysis of Spe I restricted chromosome, were detected from different inpatients' specimens during approximately 2 and a half years. The existence of such long-lasting microorganisms suggested the possibility of hospital-acquired infection caused by inadequate use of antimicrobial agents and disinfection procedures for medical tools such as bite blocks and catheters. PMID- 14743740 TI - [Relation between the prostatic tissue components and natural history, estrogen receptor]. AB - PURPOSE: There had been few reports on relations between rates of prostatic tissue components, natural history and estrogen receptor. The present study was performed to evaluate these relations. METHOD: In this study, we examined 25 patients who had benign prostatic hyperplasia(BPH) and underwent transurethral resection of the prostate(TUR-P) or suprapublic prostatectomy(SPP). According to resection volumes of BPH, patients were divided into four groups; less than 5 g of TUR-P, more than 30 g of TUR-P, less than 70 g of SPP, more than 70 g of SPP. We also examined 13 nonhyperplastic prostates that were obtained from autopsied males. Immunoenzymatic staining and computer image analysis were used to determine the mean rate of the stromal. Additionally, resection volumes of BPH were evaluated according to estrogen receptor staining. RESULTS: In nonhyperplastic glands, the stromal rates of age 15-40 were almost equal. We found that as resection volumes of BPH increased, the stromal rates were apt to decrease. However, positive rates of estrogen receptor showed no significant difference among the four groups. CONCLUSIONS: There was a trend of correlation between prostates with a decrease of stromal rate and increase of resection volume. Further studies of large populations are needed to validate this assumption. PMID- 14743741 TI - [The analysis of the sensitivity and the specificity of the mass screening system of BSE]. AB - Since the first infected case of BSE(Bovine Spongiform Encephalopathy) was identified in September, 2001 in Japan, Japanese people have been deeply concerned about safety of meat despite a mass screening system established by Japanese government. In the system all the cows for food must be examined with ELISA as screening and additionally with more accurate tests such as western blot etc. if the screened result is positive. To investigate the accuracy of the system, we calculated the number of false negatives and positives using computer simulation. Consequently, the sensitivity analysis towards ELISA showed that even if Japanese cattle were in a low risk group, a few of them are likely to be misdiagnosed as "negative" in spite of true BSE under the current screening system. Also we indicated how to improve the current system to get less false negatives. We believe the analysis of this study can contribute to risk communication with people to minimize the fear or irrationality caused by BSE epidemic. PMID- 14743742 TI - [Tumor markers in gynecological and breast cancer]. AB - Serum tumor markers are useful in diagnosis and follow-up for patients with gynecological malignancy or breast cancer. In epithelial ovarian cancer, CA125 has been identified as the most sensitive marker. Unfortunately, CA125 detection in the serum of patients with minimal malignant tumor has not been possible. Many nonmalignant conditions including endometriosis, menstruation and massive ascites may elevate the CA125, and almost 50% of patients with clear cell adenocarcinoma do not show CA125 elevated above 100 U/ml. To improve sensitivity and specificity in the diagnosis of ovarian cancer, the use of multiple tumor markers and the simultaneous use of image diagnosis should be employed. The value of tumor markers in the screening for cervical cancer and endometrial cancer has received little attention. However, the utility of serum SCC as a marker for monitoring cervical squamous cell carcinoma has been established. Since hCG is produced by gestational trophoblastic neoplasia and is a sensitive marker of trophoblastic cells in the body, patients with choriocarcinoma or invasive mole must be followed closely for this parameter. The improvement of the hCG detection technique has reduced the mortality rate from trophoblastic neoplasia. In breast cancer, many markers including CEA and CA15-3 are used, and they are reported to be useful as markers for monitoring. PMID- 14743743 TI - [Genetic tests for molecular markers of cancer]. AB - Based on recent advancements in biotechnology and molecular oncology, genetic tests for molecular markers have been widely used for diagnosis and therapeutic monitoring in patients with hematological malignancies. However, incorporation of such tests has been limited in the care of patients with non-hematopoietic malignancies. The reasons for this include difficulty in sampling of the tumor tissue, and a lack of evidence for cost-effectiveness in the test utilization. A practical approach is the use of bodily fluids, such as blood, sputum, urine and feces, as specimens for early detection and monitoring of tumor-derived nucleic acid sequences. The prospect has been raised for development of tests for individualized medicine based on genetic information such as prediction of responsiveness to chemotherapeutic agents for selection of treatment, which will be promoted by gene expression profiling using DNA microarray. PMID- 14743744 TI - [Histopathology and tumor markers]. AB - Serum tumor markers are useful for detection and diagnosis of cancer. Recent advances in cellular and molecular biology have developed procedures of immunohistochemistry including high sensitivity staining and epitope retrieval. Using the immunohistochemical analyses, we can detect various tumor markers, i.e., not only serum tumor markers (serum protein, carbohydrates), but also cellular skeletons, lymphocyte surface antigens, cytoplasmic markers, oncogene products and cell adhesion molecules. This article focuses on several immunohistochemical tumor markers. Carcinoembryonic antigen(CEA) is one of the good markers of colorectal cancer. Recent studies have demonstrated that CEA may function as a metastatic potentiator by different pathway, such as modulation of immune responses, facilitation of intercellular adhesion and cellular migration. CA19-9 (sialyl Le antigen), a member of a family of high molecular weight glycoproteins, was originally described as a gastrointestinal- and pancreatic specific tumor marker. Recent studies have demonstrated that sialyl Le is a ligand for E-selectin and may play an important role in tumor metastasis. Stromal immunoreactivities of CEA or CA19-9 were correlated with lymph node metastasis and/or vascular invasion. p53 is one of the most important tumor suppressor genes, and the p53 gene product is known to regulate cell growth and proliferation. Mutation of the p53 gene can be detected immunohistochemically as overexpression of its protein in the nucleus. Diffuse p53 immunoreactivity was a histological marker of adenocarcinoma. In conclusion, immunohistochemical tumor markers are useful for histopathological diagnosis and can be prognostic predictors of cancer. PMID- 14743745 TI - [Status quo of tumor markers and important points in medical practice]. AB - Tumor markers is a term describing substances produced by tumor cells or in response to tumor cells, and new tumor markers have been found with the progress of examination modalities. Not only Bence-Jones protein and M protein by immunoelectrophoresis, and AFP and CEA by immunochemical analysis, but also hormones, enzymes, and embryonal cancer related substances have been employed for a long time. Recently, a variety of new tumor marker tests have been developed, such as those using monoclonal antibodies that recognize carbohydrate antigens, bone metabolism markers to evaluate bone metastasis of cancer by the type I collagen metabolites, and gene markers to examine oncogenes and tumor suppressor genes by gene cloning and the PCR method. Early detection and diagnosis are keys to countermeasures against cancer, and tumor markers are regarded as a powerful tool for early detection of cancer. However, any tumor marker can be positive without cancer. In addition, problems arise because tumor markers often show negative for early cancer or even advanced cancer. As a result, no ideal tumor marker has been found. Tests of tumor markers aim to screen high-risk patients, differentiate tumor types, monitor therapeutic efficacy, and detect recurrences early. It is important to raise the diagnostic efficiency by avoiding the selection of similar markers in antigenicity, and instead combining two to three markers with little association. PMID- 14743746 TI - [Diagnostic tests approved by Ministry of Health and Welfare (November 2003)]. PMID- 14743747 TI - [The need of general internal medicine: its historical and social background]. PMID- 14743748 TI - [The role of general internal medicine in medical practice: 1. At university hospitals]. PMID- 14743749 TI - [The role of internal medicine in medical practice. 1. At general hospitals]. PMID- 14743750 TI - [The role of internal medicine in medical practice. 2. At clinics]. PMID- 14743751 TI - [The role of general internal medicine in medical education. 1. Undergraduate education]. PMID- 14743752 TI - [The role of general internal medicine in medical education. 1. Postgraduate education]. PMID- 14743753 TI - [Training of physicians who will be in charge of community health care]. PMID- 14743755 TI - [Primary care practiced in European countries and the United States]. PMID- 14743754 TI - [Research in general internal medicine]. PMID- 14743756 TI - [Future of general internal medicine in Japan]. PMID- 14743757 TI - [The current status and an overview of general internal medicine: discussion]. PMID- 14743758 TI - [Muscular sarcoidosis associated with acute renal failure due to hypercalcemia]. PMID- 14743759 TI - [Acute renal failure in a case of Waldenstrom's macroglobulinemia]. PMID- 14743760 TI - [Aortitis syndrome treated with intraaortic stenting]. PMID- 14743761 TI - [Severe hypocalcemia compatible with idiopathic hypoparathyroidism associated with psoriasis vulgaris]. PMID- 14743762 TI - [Fusobacterium necroforum sepsis]. PMID- 14743763 TI - [Fitz-Hugh-Curtis syndrome presenting spontaneous regression of ascites]. PMID- 14743764 TI - [Hypothalamic hypogonadism associated with GRH and CRH deficiencies]. PMID- 14743765 TI - [Systemic sclerosis complicated with left trigeminal neuropathy and myasthenia gravis during a period of six years after onset]. PMID- 14743766 TI - [Regenerative treatment of nervous system diseases]. PMID- 14743767 TI - [Hybrid-type artificial kidney]. PMID- 14743768 TI - [Lessons from prion disease, mad cow disease, and iatrogenic Creutzfeldt-Jakob disease]. PMID- 14743769 TI - [Discoveries through studies of influenza and the development of anti-influenza drugs]. PMID- 14743770 TI - Otolaryngologic allergy. Preface. PMID- 14743771 TI - New horizons in the management of allergy. AB - Over the last year, the understanding of early childhood allergen exposures and their impact on the development of allergy has radically changed. This article reviews these epidemiologic discoveries, although they have not yet led to therapeutic initiatives. PMID- 14743772 TI - The importance of allergy in the practice of otolaryngology. AB - Allergy in the practice of otolaryngology is not confined to patients with typical hay fever symptoms. The otolaryngologist encounters allergy as a primary cause or secondary contributor to a number of otologic, rhinologic, and laryngologic syndromes and disorders. The practitioner of otolaryngology should be prepared to recognize these situations and to address them appropriately. Training and resources are available to help the physician wishing to achieve this capability, no matter the current level of understanding and expertise. PMID- 14743773 TI - Clinical practice guidelines and specific antigen immunotherapy. AB - Allergy immunotherapy is a safe, effective treatment modality in selected patients. The length of therapy and the wide variety of patient sensitivities make it difficult to develop and test evidence-based guidelines in all areas of immunotherapy. A review of techniques and the evidence supporting them is provided in this article. PMID- 14743774 TI - Addressing the prevalence of respiratory allergy in the home environment. AB - Respiratory allergy prevalence has always depended both on genetic predisposition and specific environmental allergenic stimulation that leads to sensitization and eventual symptomatic disease. Changes brought about by modern technology that have afforded a higher quality of life have also accidentally increased the levels of many respirable and ingestible allergens in the environment. In many cases these higher levels of exposure have exceeded individual thresholds, resulting in the phenotypic expression of allergy in many individuals who were previously asymptomatic even though they were genotypically predisposed to developing allergies. Prevalence can be decreased only if susceptible populations are identified as early as possible through careful family history taking and appropriate testing for sensitivity and if exposure to selected allergens is controlled. PMID- 14743775 TI - Cost-effective pharmacotherapy for inhalant allergic rhinitis. AB - The otolaryngologist is one of the decision leaders for patients who seek to learn more about their problems of respiratory allergy. Although these patients do not have a life-threatening illness, the reduction of quality of life and performance can significantly restrict their overall sense of well being. Patients with allergic rhinitis desire the relief of the bothersome problems without other side effects. Second-generation antihistamines were introduced to reduce the significant impairment brought on by the sedation of the first generation products. Most physicians prescribe the intranasal corticosteroids as the prescription drug of first choice for most patients with chronic allergic rhinitis. Second-generation H1 receptor antagonists are better for the patient than the first-generation drugs because of the reduced side-effect profile and improved tolerance. Compliance factors certainly need to be addressed with medications that need more than once-daily dosing. Patients with only sporadic problems in season or on limited exposure are best treated with oral antihistamines, topical cromolyn, and short-term decongestant therapy. PMID- 14743776 TI - Adding allergy to your practice. AB - The ability to blend the management of allergic disease into the general treatment of head and neck disease is core to the otolaryngologist's role as regional specialist of the upper aerodigestive tract. Allergy training is now considered an integral component of the residency curriculum established by the American Board of Otolaryngology. By obtaining any needed additional training through organizations such as the AAOA and by validating that education by obtaining fellowship status in the AAOA, the postgraduate otolaryngologist who adds allergy to his or her practice will find the techniques to be safe, effective, and of great benefits to patients. PMID- 14743777 TI - Current in vivo and in vitro screens for inhalant allergy. AB - Screening for inhalant allergy requires the use of a rapid, accurate, cost effective methodology for the detection of the presence or absence of allergic responsiveness. In vivo and in vitro methods have been demonstrated to be sensitive and specific in the diagnosis of inhalant allergy, even with panels as small as seven or eight antigens. There is good concurrence between skin testing techniques and various serum measures of specific IgE, and both methods can be used effectively for screening inhalant allergy, although per-test costs for in vitro methods remain higher than comparable costs for skin testing [28]. Although the rapidity of both methods is acceptable, skin testing allows immediate observation and interpretation of test results, whereas all in vitro methods involve serum processing that delays results for hours to days. In general, both in vitro methods and epicutaneous testing with a device such as the Multi-Test II offer the clinician effective techniques in screening for inhalant allergy. Each technique has benefit and can be used in both primary care practices and in specialists' offices. The choice of the primary technique to be used for screening depends on the type of practice, the practice's familiarity with skin testing, the geographic region, and the relevant payer mix. A review of the relevant research on allergy screening suggests that the number of antigens used should be in the range of 8 to 12 and should include representative allergens from all the significant classes for the specific geographic [table: see text] region. This screen would probably contain one or two grasses, weeds, and trees, two or three molds, cat allergen, and one dust mite allergen. A sample panel is demonstrated in Box 6. It is important to recognize that the diagnosis and treatment of inhalant allergy is a fluid process. In patients who have negative allergy screens but for whom suspicion of inhalant allergy remains high, additional testing with ID techniques is sometimes indicated. This procedure might be suggested in patients for whom there is a high suspicion of mold allergy or who have unusual inhalant exposures. These individuals may represent a failure of the screening process and require additional testing. In these unusual cases, the judgment of the otolaryngic allergist is critical in guiding this further diagnostic work-up. The use of screening procedures for the diagnosis of inhalant allergy provides a rapid, accurate, and cost-effective method for evaluating patients with a history suggestive of allergic disease. Research suggests that a screening panel has strong clinometric properties and has both sensitivity and specificity in excess of 95%. It is statistically unlikely that a patient with negative findings to an allergy screen will demonstrate clinically significant inhalant allergy. PMID- 14743778 TI - Cutaneous testing for allergy diagnosis: comparison of methods in common use. AB - Practitioners who treat allergic patients must be familiar with the tests that are available to assist them in establishing the diagnosis. They must also understand the strengths and weaknesses of each of the techniques, devices, and grading systems at their disposal and that there is no criterion standard skin test. This article discusses the various techniques used in skin testing for allergy, their grading systems, and their advantages and disadvantages. PMID- 14743779 TI - In vitro testing for allergy diagnosis. AB - The measurement of specific IgE antibodies has progressed so that highly reproducible, accurate, and quality-controlled tests are available [62]. These tests can certainly assist all physicians in evaluating allergic problems and are likely to be used widely in the future. Continued research is allowing the refinement of these tests as well as increasing the understanding of allergens and allergic disease. In vitro testing techniques are major assets to allergy care. Either in vitro or skin methods are appropriate modalities for the diagnosis of allergic rhinitis [45]. When the merits of the allergy skin test and in vitro tests for diagnosis of specific allergens are compared, the following statements are considered to be correct at this time: 1. Optimally performed skin tests and in vitro tests detect IgE E antibody accurately and reproducibly. 2. Results of both tests correlate equally well with allergic signs and symptoms produced by exposure to the specifically tested allergen. 3. Both tests can be used as grounds for instituting avoidance therapy and immunotherapy efficiently and economically. PMID- 14743780 TI - Allergy management for the otolaryngologist: selecting allergenic extracts for inhalant allergy testing and immunotherapy. AB - The proper selection of allergen extracts is important in maintaining an efficient allergy practice. This article discusses pollen and mold spore selection, which may be relevant to an individual practitioner's testing survey. The selection of tree, grass, and weed pollen extracts depends on the location of the clinician's allergy practice. Allergenic extract should also be selected based on the allergenicity of the plant's pollen. Examples of pertinent allergenic trees, grass, and weed pollen are given in this article. The complexities of mold spore testing and selection are also emphasized and discussed at some length. PMID- 14743781 TI - Approaches to testing for food and chemical sensitivities. AB - Testing for food and chemical sensitivities usually becomes necessary as part of the evaluation of otolaryngology patients who have chronic illness. The more complex the patient, and the more recalcitrant the problem is to treatment, the more likely it is that allergies, and especially food or chemical sensitivities, are involved in the pathogenesis of the illness. Failure to consider all major allergen contacts, including foods and chemicals, can lead to inadequate therapy. Similarly, failure to understand total allergic and oxidant load and the effects of chemical toxicity can lead to inappropriate or ineffective treatment. Clinically, food allergies occur in two different types: immediate, anaphylactic, fixed reactions and delayed, chronic, cyclic reactions. Different test methods have been developed for the two types. Fixed food allergies can be safely and efficiently detected by in vitro specific IgE or histamine release tests. Cyclic food allergies are best detected by either oral food challenges or by the IPDFT test. Choosing the best test for a particular patient requires a clear understanding of the two food allergy types and how their clinical presentations differ. Other tests for food allergies are compared and contrasted with these primary tests. Chemical sensitivity also occurs in two different clinical types: allergic, and toxic. True allergy to chemical haptens, either type I, IgE mediated, or type IV, delayed hypersensitivity, occurs with significant frequency but is often unsuspected. Chemical toxicity can be caused by the aftereffects of an acute exposure or as a result of chronic, low-level exposure, but is even more frequently unsuspected and will not be diagnosed without a high index of suspicion. Both types of chemical sensitivity need to be addressed in any patients who have either a high allergen or chemical exposure load [105]. Either in vitro or in vivo tests can be used for chemical allergy detection; the advantages of each are outlined. Chemical toxicity screening tests are available and useful but do not detect all possible toxicants. Definitive toxic chemical tests usually require specialized laboratory facilities and expert consultation, for which possible sources are specified. The most important point in testing for food or chemical sensitivity is to be aware that food or chemical sensitivity can be contributing to a specific patient's clinical problems. Only then can appropriate investigations be undertaken to understand and then, perhaps, to intervene successfully in that illness. PMID- 14743782 TI - Allergy and the contemporary rhinologist. AB - Recently, an expanded view of inflammatory disease of the nose and paranasal sinuses, an improved understanding of the functional anatomy of this region, and the development of more sophisticated methods for examining inflammation have led to subtle changes in the field of rhinology. Careful review of the literature suggests that a number of disparate disease processes may serve as cofactors leading to the ultimate development of inflammatory nasal/paranasal sinus disease. Evaluation of cellular infiltrates and inflammatory mediators associated with various forms of rhinologic disease reveals significant overlap among many diseases that were previously thought to be separate disease processes. Further, epidemiologic studies suggest that diseases such as allergic rhinitis may be associated with the development of various forms of rhinosinusitis, and its treatment may impact the ultimate treatment for these diseases. It is becoming increasingly apparent that the contemporary rhinologist can no longer rely simply on either technical surgical skills or cognitive nonsurgical skills alone to treat these patients. Instead, to be most effective, rhinologists must synthesize diagnostic and treatment plans, using all aspects of this field of medicine. PMID- 14743783 TI - Allergy and the contemporary laryngologist. AB - The author believes that allergy plays an important role in the field of laryngology. Not every patient has significant allergic problems, but the allergic factor in laryngeal problems should not be underestimated. The insights and technology for research have never been better. Many cause-and-effect relationships have been suggested and often provide the working basis for current therapeutics. Many current models of operation need to be verified, explored further, and modified through research. It is hoped that new technologies will achieve a higher degree of sensitivity without sacrificing specificity. Better specificity is particularly needed in allergy testing and in testing thyroid and pulmonary function. The author hopes that the contemporary laryngologist/otolaryngologist will use this overview to formulate a complete and orderly approach to laryngeal problems. Because of the complexity of laryngeal problems, referral to other specialists may be necessary. The laryngologist, however, should be able to orchestrate the appropriate use of technologies and health care specialists to address these problems. PMID- 14743784 TI - Allergy and the contemporary otologist. AB - Allergy may affect the outer, the middle, or the inner ear. Although the otologic manifestations of allergy are not by themselves diagnostic, the history, including family history and associated symptoms in other target organs, will often help lead to the correct diagnosis and institution of therapy. Patients with significant and chronic symptoms, including those with labyrinthine symptoms of allergy, respond well to specific immunotherapy or dietary elimination. PMID- 14743785 TI - Anaphylaxis. AB - Anaphylaxis is an amplified, harmful immunologic reaction that occurs after re exposure to an antigen to which an organism has become sensitive. True anaphylaxis is a systemic reaction caused by antigen-specific cross-linking of IgE molecules or complement proteins on the surface of tissue mast cells and peripheral blood basophils, resulting in the immediate release of potent mediators. Immediate systemic reactions that resemble anaphylaxis but are not caused by an IgE-mediated immune response are referred to as anaphylactoid reactions. It is important for physicians, especially those who treat allergies, to understand the pathophysiology, know the treatment for, and recognize the clinical signs of anaphylaxis. PMID- 14743786 TI - Contact and chemical sensitivities in the hospital environment. AB - As surgeons, otolaryngologists tend to most be interested in operative procedures and leave the hospital environment to the care of administrators and the nursing staff. Given the dangers that are present, it would seem prudent to spend some time considering the agents that used in patient care and in operating suites, to minimize the risk to patients and co-workers. PMID- 14743787 TI - Prevention and treatment in Eastern Europe. PMID- 14743788 TI - Finding a voice in Vietnam. PMID- 14743790 TI - Tales of war. PMID- 14743789 TI - Botswana--the challenges of fighting HIV/AIDS. PMID- 14743792 TI - Waiting in the wings: more new drugs. PMID- 14743791 TI - HIV treatment series II. Part 3 of 4. Managing side effects. PMID- 14743793 TI - Tuberculosis: not a thing of the past. PMID- 14743794 TI - HCV/HIV co-infection III--a patient's perspective. PMID- 14743795 TI - I love my Androgel. PMID- 14743797 TI - HIV and personal finance. Where do we go from here? PMID- 14743796 TI - Human rights in Vancouver: do injection drug users have a friend in city hall? AB - In april 2003, the Vancouver Police Department (VPD) initiated a crackdown in the city's Downtown Eastside, an area frequented by drug users and the site of one of the developed world's worst AIDS epidemics. Human Rights Watch (HRW) visited the city and issued a report documenting first-hand accounts of unnecessary use of force by police officers and other human rights abuses. In this article, HRW staffers Joanne Csete and Jonathan Cohen describe how the initial euphoria that greeted the election of Vancouver Mayor Larry Campbell has worn off; how a crackdown that was supposed to be aimed at drug traffickers had the effect of driving drug users away from health and harm-reduction services; and how both the VPD and city tried to discredit the HRW report. Finally, the authors discuss how concerns about the VPD have led to official complaints being filed, and they question whether police forces should be allowed to investigate themselves. PMID- 14743798 TI - Disclosing HIV in the workplace. One personal perspective from the corporate world. PMID- 14743799 TI - Great expectations. "At first I had high hopes for this new job, but now I'm wondering where it all went wrong". PMID- 14743800 TI - Stress reduction. Healing of all kinds requires a state of rest, which may be impossible with chronic stress. PMID- 14743801 TI - China: government issues licences for three antiretrovirals. AB - The Chinese government has approved the production and sale of at least three generic antiretroviral drugs used in treating people living with HIV/AIDS. The first applications were filed in late 2001 and approved in August and September 2002. PMID- 14743802 TI - Kenya: legislative mendments ease imports of generic drugs. PMID- 14743803 TI - South Africa: voluntary licence to generic drug company for nevirapine. PMID- 14743804 TI - South Africa: TAC, COSATU and allies file complaint of excessive pricing. AB - On 19 September 2002, the Treatment Action Campaign (TAC), the Council of South African Trade Unions (COSATU), the Chemical, Energy, Paper, Printing, Wood and Allied Workers' Union (CEPPWA), and eight individuals (health-care workers and people living with HIV/AIDS) launched a complaint with the country's Competition Commission against two major transnational pharmaceutical companies. TAC and its allies allege that GlaxoSmithKline (GSK) has engaged in excessive pricing for its antiretroviral drugs Retrovir (zidovudine or AZT), 3TC (lamivudine), and Combivir (AZT/lamivudine), and that Boeringer Ingelheim (BI) has engaged in excessive pricing of its antiretroviral drug Viramune (nevirapine). PMID- 14743805 TI - WTO: US and Argentina settle dispute over patents and data protection. AB - In May 2000, supplementing an earlier complaint filed in May 1999, the US filed a complaint against Argentina, alleging that its patent laws violate the World Trade Organization's Agreement on Trade-Related Aspects of Intellectual Property (the TRIPS Agreement). The gist of the US complaint was that Argentina's law failed to provide: (1) adequate protection against "unfair" commercial use of undisclosed test data submitted in order to get market approval of pharmaceutical products; (2) certain safeguards for compulsory licences on an invention granted on the basis of inadequate working by the patent holder; and (3) adequate measures to prevent infringements of patent rights. The US also alleged that Argentina denies certain exclusive rights of patent holders, such as the exclusive right to import the patented product into the country. At the end of May 2002, the US and Argentina notified the WTO that they had reached a "mutually agreed solution," without prejudice to their respective rights and obligations under WTO agreements, and the US has withdrawn its complaint. PMID- 14743806 TI - Inter-American Commission on Human Rights hold hearing on access to treatment in Latin America and the Caribbean. AB - In a historic development, non-governmental HIV/AIDS organizations in Latin America and the Caribbean presented a joint report on access to comprehensive care, including antiretroviral (ARV) drugs, to the Inter-American Commission on Human Rights on 16 October 2002. PMID- 14743807 TI - BC court orders government to pay for treatment. AB - On 9 October 2002, the British Columbia Court of Appeal upheld a ruling of a BC court that the BC government must not discriminate against a disabled and disadvantaged group when choosing what medical treatments it will fund. The Court of Appeal ordered the BC government to pay for a particular form of treatment. The case is significant in the context of HIV/AIDS because it could lend support to arguments that a government must make appropriate accommodation for the health care needs of other disabled and marginalized groups--for example, safe injection supervision for the treatment of addiction. PMID- 14743808 TI - Discrimination in same-sex survivor amendments to the Canada Pension Plan. AB - On 31 July 2002, the British Columbia Supreme Court granted certification of a class proceeding involving same-sex surviving spouses who were denied survivor's benefits under the Canada Pension Plan (CPP) due to their sexual orientation. On 10 July 2002, a similar proceeding in Ontario survived a motion to strike by the government and is headed to trial. If successful, thousands of gay men and women whose same-sex partners died between 17 April 1985 and 1 January 1998 may finally be eligible for monthly survivor's benefits. PMID- 14743809 TI - Supreme Court denies leave to appeal in tainted-blood case. PMID- 14743810 TI - Supreme Court denies leave to appeal in medical marijuana charter challenge. AB - As previously reported in this publication, in January 2002 the Ontario Court of Appeal denied Jim Wakeford's claim that Canada's laws prohibiting marijuana possession and cultivation infringe his constitutional rights to liberty and security of the person. On 22 November 2002 the Supreme Court of Canada announced that it would not hear his appeal from that decision. PMID- 14743811 TI - Vancouver Island Compassion Club president receives absolute discharge. AB - On 5 July 2002, the BC Provincial Court released written reasons for sentencing in R v Lucas. The president of the Vancouver Island Compassion Club had been arrested and charged with possession for the purpose of tracking after police searched the club and his home and seized less than three kilograms of marijuana. PMID- 14743812 TI - BC Court of Appeal refuses to reduce sentence for HIV-positive prisoner. AB - On 9 May 2002, the BC Court of Appeal released its decision in a prisoner's sentencing appeal where the Crown and the defence agreed that the original sentence was illegal, and differed in their positions as to the appropriate sentence by only one day. That one day meant the difference between serving the sentence in a provincial jail as opposed to a federal penitentiary. PMID- 14743813 TI - Criminal law and HIV transmission/exposure: more new cases. PMID- 14743814 TI - Barcelona 2002: law, ethics, and human rights. An advocacy and political conference produces a consensus: effective action is urgently required. AB - From the deep sense of frustration about the gap between what is possible and what is actually happening, a clear consensus emerged at the XIV International AIDS Conference that effective action is urgently required. This article is based on a presentation on 12 July 2002, the last day of the conference, by Terje Anderson, the rapporteur for Track G. The article presents a summary of the discussions in Track G on topics such as advocacy strategies, the use of the law, the use of a human rights framework and approach, the role of people living with HIV/AIDS, and the need to mobilize sufficient resources. The article states and then critically examines some of the consensus positions that emerged from the conference--specifically, the goal to have three million people on antiretroviral therapy by 2005; the notion that the debate around prevention versus care is over; and the idea that the key issue is no longer what we do, but how to secure the commitment and the resources to do it. The article states that the fight against HIV/AIDS must be fought on a political plane, and that it is the responsibility of everyone working in AIDS to engage our leaders. The article concludes by asking whether we really have the courage and the perseverance to turn our knowledge and our commitment into action. PMID- 14743815 TI - Barcelona 2002: law, ethics, and human rights. Global battle cry: health is a right, not a commodity. AB - Health is a fundamental right, not a commodity to be sold at a profit, argues Irene Fernandez in the second Jonathan Mann Memorial Lecture delivered on 8 July 2002 to the XIV International AIDS Conference in Barcelona. Ms Fernandez had to obtain a special permit from the Malaysian government to attend the Conference because she is on trial for having publicly released information about abuse, torture, illness, corruption, and death in Malaysian detention camps for migrants. This article, based on Ms Fernandez' presentation, describes how the policies of the rich world have failed the poor world. According to Ms Fernandez, the policies of globalization and privatization of health care have hindered the ability of developing countries to respond to the HIV/AIDS epidemic. The article decries the hypocrisy of the industrialized nations in increasing subsidies to farmers while demanding that the developing world open its doors to Western goods. It points out that the rich nations have failed to live up their foreign aid commitments. The article concludes that these commitments--and the other promises made in the last few years, such as those in the United Nations' Declaration of Commitment on HIV/AIDS--can only become a reality if they are translated into action. PMID- 14743816 TI - Barcelona 2002: law, ethics, and human rights. The need for harm-reduction approaches in Eastern Europe and the former Soviet Union. AB - In a region where HIV is spread primarily by injection drug use, harm-reduction strategies must be the mainstay of prevention efforts. In her plenary presentation to the XIV International Conference on AIDS on 9 July 2002, Kasia Malinowska-Sempruch asserts that if the world does not turn its attention to the emerging and exploding epidemic in Eastern Europe and the former Soviet Union, the region will suffer the same fate as Africa. The presentation explains that while their economies continue to struggle, the countries in the region have seen their public health systems crumble in the face of the twin health crises of injection drug use and HIV infection. The presentation demonstrates how current repressive laws and practices with respect to drug use serve only to fuel the epidemic. It describes harm-reduction approaches (such as needle exchanges and drug-treatment programs) and provides examples of how NGOs in the region have been attempting to implement such approaches, often with little or no support from governments. Finally, the presentation outlines other measures required to respond to the epidemic in the region, including ensuring that people infected with HIV can access care, treatment, and support services. PMID- 14743817 TI - Barcelona 2002: law, ethics, and human rights. Using the law to improve access to treatments. AB - The XIII International AIDS Conference in Durban, South Africa in July 2000 focused worldwide attention on the problem of accessing treatments in developing countries. In the interim, thanks to the work of activists - from demonstrations to court cases, and from acts of public courage by people living with HIV/AIDS to ongoing lobbying of politicians and trade negotiators - some very significant developments have occurred. But the reality is that the vast majority of people living with HIV/AIDS still lack access to affordable, quality medicines. This article, a summary of a paper presented at "Putting Third First: Vaccines, Access to Treatments and the Law," a satellite meeting held at Barcelona on 5 July 2002 and organized by the Canadian HIV/AIDS Legal Network, the AIDS Law Project, South Africa, and the Lawyers Collective HIV/AIDS Unit, India, explores three approaches for improving access. In the first part, Richard Elliott provides an overview of the state of the right to health as embodied in international human rights law; comments on the experience to date in litigating claims to the right to health; and identifies potential strategies activists can adopt to advance recognition of the right to health. In the second part, Sharan Parmar and Vivek Divan describe price-control and drug-financing mechanisms used by industrialized countries to increase the affordability of medicines; and discuss how some of these mechanisms could be adapted for use in developing countries. Finally, Jonathan Berger describes the use of litigation in the courts by the Treatment Action Campaign in South Africa. PMID- 14743818 TI - Barcelona 2002: law, ethics, and human rights. Introducing ARVs in resource-poor settings: challenges and consequences. AB - In most industrialized countries, AIDS mortality has plunged sharply with the advent of antiretroviral therapy. Yet the majority of people living with HIV/AIDS do not have access to modern HIV care, and some experts have argued that introducing such therapy is not a ranking priority. In this article, which is based on a plenary presentation at the XIV International AIDS Conference, Paul Farmer presents the experience of an integrated HIV prevention and care project in rural Haiti, and explores the challenges to national AIDS programs and other bodies in the "least developed" countries as more resources are made available for HIV prevention and care. PMID- 14743819 TI - Barcelona 2002: law, ethics, and human rights. "I have no watch, but I haven't missed a dose". AB - Anyone who was fortunate enough to hear Fred Minandi speak in Barcelona must have been moved by what he had to say. Mr Minandi is a person living with HIV/AIDS from Chiradzulu, Malawi, who spoke at Time to Treat: Transforming AIDS Treatment from Right to Reality, a satellite meeting of the XIV International Conference on AIDS held on 7 July 2002, and sponsored by Medecins Sans Frontieres (MSF) and the Health Global Access Project. His presentation is a passionate, first-hand account of how antiretroviral therapy has transformed lives, how it has given hope to people, how it has helped to reduce the stigma against AIDS in his community, and how it has contributed to prevention efforts. PMID- 14743820 TI - Barcelona 2002: law, ethics, and human rights. Advancing research and access to HIV vaccines: a framework for action. AB - In light of the continuing spread of HIV infection and the devastating impact of the disease on lives, communities, and economies, particularly in the developing world, the investment in new treatments, vaccines, and microbicides has clearly been inadequate. Efforts must be intensified to develop effective HIV vaccines and to ensure that they are accessible to people in all parts of the world. This article is a summary of a paper by Sam Avrett presented at "Putting Third First: Vaccines, Access to Treatments and the Law," a satellite meeting held at Barcelona on 5 July 2002 and organized by the Canadian HIV/AIDS Legal Network, the AIDS Law Project, South Africa, and the Lawyers Collective HIV/AIDS Unit, India. In the article, Avrett calls for immediate action to increase commitment and funding for HIV vaccines, enhance public support and involvement, accelerate vaccine development, and plan for the eventual delivery of the vaccines. The article briefly outlines steps that governments need to take to implement each of these objectives. The article also provides a menu of potential actions for vaccine advocates to consider as they lobby governments. PMID- 14743821 TI - Barcelona 2002: law, ethics, and human rights. Juggling individual and collective concerns with respect to HIV/AIDS policies. AB - Efforts to prevent the spread of HIV infection sometimes give rise to tensions between individual and collective rights. This article, based on a presentation by Nelson Varas-Diaz (abstract TuOrG1171), explores these tensions in the context of the laws and policies of eight Latin American countries: Costa Rica, Dominican Republic, Ecuador, Guatemala, Honduras, Nicaragua, Panama, and Puerto Rico. The article describes five elements of the response to HIV/AIDS in which tensions between individual and collective rights have surfaced: the participation of people living with HIV/AIDS on national commissions; the ability of HIV-positive persons to access antiretroviral medications; HIV-antibody testing practices; the confidentiality of health information; and the rights and duties of people living with HIV/AIDS. The article concludes that the success of programs designed to prevent the spread of HIV infection depends on the ability of societies and governments to balance the tensions between individual and collective rights. PMID- 14743822 TI - Barcelona 2002: law, ethics, and human rights. Toward a more meaningful involvement of people living with HIV/AIDS. AB - All too often, the involvement of people living with HIV/AIDS in AIDS NGOs is tokenistic rather than meaningful. This article, which is based on a presentation by Chistophe Cornu (abstract WeOrG1292), demonstrates that, under the right conditions, the involvement of people living with HIV/AIDS can be beneficial both for the individuals and for the NGOs. The article describes the beneficial impacts of involvement observed in a study conducted in Burkina Faso, Ecuador, India, and Zambia. It also discusses the harmful effects of involvement, for individuals as well as NGOs, identified by study participants. The article concludes with a series of recommendations for NGOs to maximize the benefits of involvement. PMID- 14743823 TI - Barcelona 2002: law, ethics, and human rights. HIV testing for peacekeeping forces: legal and human rights issues. AB - In 2001, the United Nations Security Council established an Expert Panel to study the issue of whether the UN should institute HIV testing of peacekeeping personnel. This article, based on a 9 July 2002 presentation to the XIV International AIDS Conference (abstract TuOrG1173), reports on the findings of a paper prepared for the Expert Panel by the Canadian HIV/AIDS Legal Network. The paper examined whether it is permissible for the UN to implement mandatory HIV testing of its peacekeeping personnel, and whether HIV-positive UN peacekeeping personnel should be excluded or restricted from service on the basis of their HIV status or HIV disease progression. The article describes some of the court cases in which these issues have been considered; discusses the importance of analyzing such issues in the context of a human rights-based approach to the pandemic; and formulates a series of key principles for guiding UN decision-making. The article concludes that a policy of mandatory HIV testing for all UN peacekeeping personnel cannot be justified on the basis that it is required in order to assess their physical and mental capacity for service; that HIV-positive peacekeeping personnel cannot be excluded from service based on their HIV status alone, but only on their ability to perform their duties; and that the UN cannot resort to mandatory HIV testing for all UN peacekeeping personnel to protect the health and safety of HIV-negative personnel unless it can demonstrate that alternatives to such a policy would not reduce the risk sufficiently. In the end, the Expert Panel unanimously rejected mandatory testing and instead endorsed voluntary HIV counselling and testing for UN peacekeeping personnel. PMID- 14743824 TI - Barcelona 2002: law, ethics, and human rights. HIV/AIDS-related stigma and discrimination: a conceptual framework. AB - Only if we understand where HIV/AIDS-related stigma and discrimination come from, and how they are connected to broader social inequalities and the denial of fundamental human rights, can we develop effective strategies to combat them. This article is a much-condensed version of a keynote presentation given at "Meeting the Stigma Challenge: New Paradigms for Civil Society," a satellite meeting held in Barcelona on 8 July 2002, and sponsored by the Joint United Nations Programme on HIV/AIDS (UNAIDS). The presentation described the conceptual framework underpinning the 2002-2003 World AIDS Campaign, whose theme is "HIV/AIDS-Related Stigma and Discrimination," and whose slogan is "Live and Let Live." In this article, Peter Aggleton provides a conceptual overview of the relationship between the stigma and discrimination associated with HIV and AIDS and the human rights violations that ensue from them, with the goal of demonstrating the interconnectedness of these concerns. He also provides some examples of concrete steps that can be taken to counter the stigma, discrimination, and human rights violations. PMID- 14743825 TI - Traffic volumes and respiratory health care utilization among residents in close proximity to the Peace Bridge before and after September 11, 2001. AB - A recent study based on data over a 10-year period (1991-2000) showed a positive association between health care utilization and prevalence of asthma, and commercial traffic at a U.S.-Canada border crossing. We wanted to determine whether decreases in total traffic would also be associated with decreases in health care utilization for respiratory illnesses. Following September 11, 2001, there was a 50% drop in total traffic at the Peace Bridge border crossing point between Buffalo, New York and Fort Erie, Ontario, Canada. To investigate the impact of such a traffic decline on health care utilization for respiratory illnesses, weekly respiratory admissions to Kaleida Health System, Western New York's largest health care provider were analyzed according to ICD9CM classification and compared with total weekly traffic volumes for 3-month periods in 2000 and 2001 (August, September, and October). The total number of patients admitted to hospital or seen in emergency departments for respiratory illnesses during the 3-month periods of both years was 5288. A 50% drop in total traffic following Labor Day and September 11, 2001, from week 4 to week 7 was found to be statistically significant (p = 0.031) when a one-way ANOVA was performed. Likewise, the drop in total respiratory cases approached statistical significance (p = 0.052) when a one-way ANOVA was conducted. The results suggest an association between decrease in traffic volumes with decrease in health care utilization for respiratory diseases. These results suggest that current levels of traffic may be impacting on the respiratory health of residents in the nearby community. PMID- 14743826 TI - [Reaction of IAMS on broadcast TROS Radar about animal experimentation. IAMS does animal friendly research]. PMID- 14743827 TI - The role of special group article in ancient chinese medical prescription. AB - In the traditional medicine of the East, a prescription to cure diseases consisted of plural sorts of active natural products. Every article of those products has native features respectively, but each article was supposed to carry out fixed roles in the prescription. An article contains generally some sorts of active compounds, therefore it could be able to divide the role from the chemical, medicinal natures of those compounds. For example two groups of articles could be able to list up, each of which was supposed to carry out the proper role in various prescriptions. G. I. Articles were expected to adjust the whole of a prescription, to aid other articles on the induction of them in the inner part and on the removal of waste materials, with their mild, innocuous (or fragrant) natures GUI, JIAO, JIANG, GANCAO, RENCAN, GEGEN, YUANZHI. G. II. Articles were expected to carry out the leading role for cure even on serious diseases, to act on the cause of illness actively, and to remove the harmful product due to diseases, with their strong physical operations WUTOU , LANGDANG, DAHUANG, BADOU, SHEXIANG, SHAOYAO, MAHUANG. The conclusion - many compounds in those articles have physiological and medicinal effects for human body originated in their molecular structures, and these facts could endorse above expectations. PMID- 14743828 TI - The effects of speech output technology in the learning of graphic symbols. AB - The effects of auditory stimuli in the form of synthetic speech output on the learning of graphic symbols were evaluated. Three adults with severe to profound mental retardation and communication impairments were taught to point to lexigrams when presented with words under two conditions. In the first condition, participants used a voice output communication aid to receive synthetic speech as antecedent and consequent stimuli. In the second condition, with a nonelectronic communications board, participants did not receive synthetic speech. A parallel treatments design was used to evaluate the effects of the synthetic speech output as an added component of the augmentative and alternative communication system. The 3 participants reached criterion when not provided with the auditory stimuli. Although 2 participants also reached criterion when not provided with the auditory stimuli, the addition of auditory stimuli resulted in more efficient learning and a decreased error rate. Maintenance results, however, indicated no differences between conditions. Finding suggest that auditory stimuli in the form of synthetic speech contribute to the efficient acquisition of graphic communication symbols. PMID- 14743829 TI - Meals at medical specialty society annual meetings: a preliminary assessment. AB - Little isd known about how meals are chosen for medical meetings. We surveyed the annual meeting planners for 20 major specialty societies. Thirteen (65%) responded; all were currently planning their next meeting. Attendance in 2000 was reported at 113,477 physicians, with 2 million planned meals and snacks. No physician was named as responsible for food choices; the meeting planner and staff were primarily responsible for deciding what food to serve, excluding exhibit halls. Twelve (92%) respondents rated "available budget" as the most important factor. "Nutritional guidelines" were rated "very important" by eight of 13 (63%). However, no specific nutritional guidelines could be identified by any planner. All respondents indicated that members would attend a meeting if "healthy" food were the only option. For 2000, 100% of respondents indicated that for each lunch and for each dinner, a dessert had been included. No annual meeting and no planned 2001 meeting excluded potato chips, snack mixes, or candies at breaks; soda pop was offered at each break. Most respondents (89%) relied on a concluding questionnaire about the meeting facilities to evaluate the food. Respondents reported no difference in charges for "special meals," including vegetarian and kosher meals. Physicians may be unaware that some food served at medical meetings may impair learning, with excessive calorie, fat, and carbohydrate consumption. Small changes can improve the quality of food and beverages selected, without increased cost, and provide choices that conform to national nutritional guidelines. Medical meetings should serve flavorful, healthful food. PMID- 14743830 TI - Molecular and cellular regulation of the gastric proton pump. AB - The gastric H+, K+-ATPase is a proton pump that is responsible for gastric acid secretion and that actively transports protons and K+ ions in opposite directions to generate in excess of a million-fold gradient across the membrane under physiological conditions. This pump is also a target molecule of proton pump inhibitors which are used for the clinical treatment of hyperacidity. In this review, we wish to summarize the molecular regulation of this pump based on mutational studies, particularly those used for the identification of binding sites for cations and specific inhibitors. Recent reports by Toyoshima et al (2000, 2002) presented precise three-dimensional (3-D) structures of the sarcoplasmic reticulum (SR) Ca2+-ATPase, which belongs to the same family as the gastric H+, K+-ATPase. We have studied the structure-function relationships for the gastric H+, K+-ATPase using 3-D structures constructed by homology modeling of the related SR Ca2+-ATPase, which was used as a template molecule. We also discuss in this review, the regulation of cell surface expression and synthesis control of the gastric proton pump. PMID- 14743831 TI - Accuracy of peripheral quantitative computed tomography (pQCT) for assessing area and density of mouse cortical bone. AB - Peripheral quantitative computed tomography (pQCT) is increasingly used for measurement of cortical bone geometry and density in mice. We evaluated the accuracy of pQCT for area and density measurements of thin-walled aluminum phantoms and mouse femora. Aluminum tubes with varying wall thicknesses and femora from 1- to 6-month-old C3H/HeJ (C3H) and C57B1/6J (B6) mice (average cortical thickness 0.14-0.29 mm) were scanned at 70- or 90-microm resolution. pQCT values of area were compared to optical values determined after sectioning, while pQCT density (vBMD) was compared to solid aluminum density or correlated to bone ash content. For the aluminum phantoms, the error in pQCT area and density depended strongly on wall thickness, and density was consistently underestimated. For mouse femora, threshold values were found that produced zero error in bone area for each strain and age group, although the optimal threshold differed between groups. pQCT vBMD correlated strongly with ash content (r2=0.7), although the regression equations differed between strains and the magnitude of the inter strain difference in vBMD was fourfold greater than the difference in ash content. This finding suggests that pQCT can overestimate the differences in volumetric mineral density between inbred mouse strains whose bones are of different thickness (e.g., C3H vs. B6). In conclusion, both area and density values obtained by pQCT depend strongly on specimen thickness, consistent with a partial volume averaging artifact. Investigators using pQCT to assess cortical bones in mice should be aware of the potential for cortical thickness-dependent errors. PMID- 14743833 TI - Iliopsoas muscle injury during a femoral artery puncture: benign but disabling. PMID- 14743834 TI - D-I-V-O-R-C-E needn't spell financial ruin. PMID- 14743835 TI - Forecasting the impact of genetics and nursing. PMID- 14743836 TI - Ehlers-Danlos Syndrome, classical type: case management. AB - As the field of genetics expands, there will be more need for health care professionals to possess basic knowledge of genetic conditions and patterns of inheritance to assist their patients and to make the proper referrals. Ehlers Danlos Syndrome (EDS) is a group of genetic connective tissue disorders that affects approximately 1 in 5,000 live births, including males and females of all racial and ethnic groups. The main characteristics of EDS are skin hyperextensibility, tissue fragility, and joint hypermobility. Diagnosis of EDS is often difficult due to the complexity of symptoms and lack of specific genetic tests. However, once a diagnosis is suspected or confirmed, nurses play a vital role in assisting the patient and family to manage the disorder. This article describes EDS symptoms and genetic basis, provides suggestions for management, and reviews resources available for health care providers, families, and patients with EDS. PMID- 14743837 TI - Anticipatory guidance for parents of Prader-Willi children. AB - Prader-Willi syndrome (PWS) is a complex genetic disorder. It is characterized by hypotonia, short stature, hypogonadism, mental retardation, behavioral problems, and hyperphagia, which result in excessive obesity (Lindgren et al., 2000). The abnormal body composition resembles children seen with growth hormone deficiency (Carrel & Allen, 2001) . The dysmorphic features characteristic of PWS include a narrow forehead, a broad nasal bridge, slightly up slanting almond-shaped palpebral fissures, a down turned mouth with a thin upper lip, and narrow hands and feet (Martin et al., 1998). Management of children with PWS requires an ongoing multidisciplinary approach. The delivery of care includes assistance from geneticists, nutritionists, internists, endocrinologists, physical therapists, and psychologists to meet the medical, developmental, behavioral, and social needs. The focus of the nurse practitioner should include assisting the family in the management of these complex patients throughout their childhood. PMID- 14743838 TI - Cultural intersections in the care of Mexican American children with chronic conditions. AB - Cultural intersections occur whenever people from different cultures meet. Intracultural intersections may also occur between people with a similar heritage but different life experiences. Cultural intersections can present challenges in communication and understanding between people with differing values and priorities. At the same time, they offer opportunities for growth, illuminate universal suffering, and foster compassion. This article describes cultural intersections experienced by Mexican American parents of children with chronic conditions. These intersections affect relationships between parents and among parents, nurses, and other health care providers, sometimes providing unexpected insights for parents about their own lives and experiences. Nurses who are aware of cultural intersections in their practices can enhance their appreciation for the diversity existing within and between cultural groups, while focusing on the universal goal of providing excellent care for children with chronic conditions. PMID- 14743839 TI - Pediatric management problems. Sickle cell disease. PMID- 14743840 TI - Genetic link to unusual cardiac diagnosis. PMID- 14743841 TI - Thalassemia: the facts and the controversies. AB - In the United States, we continue to struggle with and make progress in the treatment of thalassemia. As our culture becomes more diverse, we will likely see more thalassemia in our pediatric populations. Currently, all newborns in the United States are screened for hemoglobinopathies. If a newborn screen returns with large amounts of fetal hemoglobin, alpha hemoglobin, or hemoglobin E, further investigation for thalassemia takes place. This article refreshes knowledge for those not immediately familiar with the red blood cell diseases and presents ethical controversies in care. The Cooley's Anemia Foundation Web site (www.cooleysanemia.org) provides a wealth of information for those with additional interests. PMID- 14743842 TI - Management and prevention strategies for respiratory syncytial virus (RSV) bronchiolitis in infants and young children: a review of evidence-based practice interventions. AB - Respiratory syncytial virus (RSV) is the most common cause of bronchiolitis and pneumonia among infants and children. The purpose of this paper is to review the epidemiology, etiology, pathophysiology, clinical manifestations, risk factors, and assessment of RSV infection in infants and young children. There is a lack of consensus regarding the optimal treatment for children with RSV infection. Bronchodilators, racemic epinephrine, inhaled and systemic corticosteroids, RSV immunoglobulin (RSV-IG), and ribavirin have all been used for treatment of children with RSV infection. A review of current research indicates supportive and symptomatic management should be the mainstay of treatment. Ultimately, prevention of infection through education and immunotherapy is the key to reducing the morbidity and mortality associated with RSV bronchiolitis. PMID- 14743843 TI - Great expectations: a position description for parents as caregivers: Part I. AB - Parents caring for a child with a chronic condition must attend to a myriad of day-to-day management responsibilities and activities. This two-part series examines parental roles and responsibilities in detail. Part I will review both the adult and pediatric family caregiving literature. It will also describe four major caregiving responsibilities with associated tasks in both direct and indirect illness-related care, which parents are expected to perform when caring for a child with a chronic condition. These are: (a) managing the illness; (b) identifying, accessing, and coordinating resources; (c) maintaining the family unit; and (d) maintaining self. Part II (to be published in the next issue of Pediatric Nursing) will present a multifaceted list of parent caregiving management responsibilities and associated activities drawn from the literature. Clinical nursing implications associated with caregiver responsibilities and tasks will also be addressed. PMID- 14743844 TI - Infant with iron deficiency anemia. PMID- 14743845 TI - Preventing meningitis. PMID- 14743846 TI - Learning intuition--less college and more kindergarten: the leader's challenge. AB - Learning to think instinctually and to depend on our intuition means, for many of us, going back to kindergarten and focusing more on our instinctual side to balance the linear approach that we've learned in many college and business school courses. It also means building time into our lives for reflection and allowing our brains the time to reorganize the events of our lives into patterns that we will recognize in the future. The process of the incubating experience and data in our minds over time will build the intuition and instincts that will help us all to learn how to be great leaders (Gardner, 1995). We can learn much from the training of clinicians in pattern recognition and intuitive analysis of clinical situations. Many of us who were clinicians first understand the intuitive side of clinical practice and of administrative practice because we were educated in both worlds. We have the advantage of knowing the concepts of intuitive thinking and must remember to apply those concepts daily to our practices in leadership. Toffler said "You can use all the quantitative data you can get, but you still have to distrust it and use your own intelligence and judgment" (Boone, 1999, p. 80). As we acquire more experience and wisdom, we learn to trust our own intelligence and judgment. What we came to know in kindergarten is as valuable as the analytical knowledge we have since acquired. In leadership, we will do well to remember our clinical roots and to continually hone our skills in intuition and thinking with our gut. PMID- 14743847 TI - Assessing ventricular function for risk stratification. PMID- 14743848 TI - Optimizing adjunctive antithrombotic therapy in the treatment of acute myocardial infarction: a role for low-molecular-weight heparin. AB - Thrombotic complications account for a large proportion of in-hospital deaths from acute myocardial infarction (MI). Although thrombolytic therapy has greatly improved clinical outcomes following MI, thrombin released during clot lysis has a prothrombotic effect, and the thrombolytic agents themselves may directly activate platelets. Antithrombotic therapy as an adjunct to thrombolysis improves the speed and extent of artery recanalization and reduces the incidence of secondary ischemic complications. The current treatment standard is unfractionated heparin (UFH) administered intravenously for 24-48 h. However, UFH has not been unequivocally shown to improve outcomes in large-scale, randomized clinical trials, and shows no evidence of benefit when used as an adjunct to streptokinase-based thrombolysis. Unfractionated heparin also has several clinical and practical disadvantages, such as the need for coagulation monitoring, difficulties attaining a stable and reliable anticoagulant effect, and the risk of hemorrhagic side effects. Low-molecular-weight heparin (LMWH) represents a safe and effective alternative antithrombotic therapy, with a stable and predictable anticoagulant effect, potential for use in combination with either fibrin-specific or streptokinase-based thrombolysis, no need for anticoagulation monitoring, and a low risk of hemorrhagic and other heparin related complications. Several randomized clinical trials have shown that adjunctive LMWH is at least as effective as UFH in the acute phase of MI, is associated with fewer in-hospital recurrent ischemic events, and has an acceptable safety profile. PMID- 14743849 TI - Atheromatous plaque cap thickness can be determined by quantitative color analysis during angioscopy: implications for identifying the vulnerable plaque. AB - BACKGROUND: Coronary angioscopy in acute myocardial infarction has frequently revealed disrupted yellow lesions. Furthermore, postmortem studies have demonstrated that these lesions have thin collagenous caps with underlying lipid rich cores. HYPOTHESIS: We hypothesized that the yellow color is due to visualization of reflected light from the lipid-rich yellow core through a thin fibrous cap. Thus, quantification of yellow color saturation may estimate plaque cap thickness and identify vulnerable plaques. METHODS: To test this hypothesis, the feasibility of detecting cap thickness was tested using both a model of lipid rich plaque and human atherosclerotic plaque. The model was constructed by injecting a yellow beta-carotene-lipid emulsion subendothelially into normal bovine aorta. Human plaque was obtained from cadaver aorta. Digitized images were obtained by angioscopy, and percent yellow saturation was analyzed using a custom computer program. Plaque cap thickness was measured by planimetry of digitized images on stained tissue sections. Percent yellow saturation was then correlated with plaque cap thickness. RESULTS: In the bovine model, plaque cap thickness and percent yellow saturation correlated inversely (r2 = 0.91; p = 0.0001). In human plaques, yellow saturation was significantly greater in atheromatous than in white plaques (p < 0.0004). Also, there was a high correlation between plaque cap thickness and yellow saturation at various angles of view between 40 degrees and 90 degrees, the greatest between 50 degrees and 80 degrees (r2 = 0.75 to 0.88). CONCLUSION: Plaque cap thickness is a determinant of plaque color, and this can be assessed by quantitative colorimetry. Thus, plaque color by angioscopy may be useful for detecting vulnerable plaques. PMID- 14743850 TI - Marked low-density lipoprotein cholesterol reduction below current national cholesterol education program targets provides the greatest reduction in carotid atherosclerosis. AB - BACKGROUND: Current National Cholesterol Education Program (NCEP) guidelines recognize low-density lipoprotein cholesterol (LDL-C) below 100 mg/dl as an optimal level. Evidence supporting this is scant. Both LDL-C and C reactive protein (CRP) are known correlates of atherosclerosis progression. HYPOTHESIS: We examined the effect of final LDL-C and CRP obtained with statin therapy on carotid intima-media thickness (CIMT), a valid surrogate for clinical benefit of lipid-lowering therapies. METHODS: In a randomized, single-center trial, 161 patients were assigned to statin therapy of different potencies (pravastatin 40 mg, n = 82; atorvastatin 80 mg, n = 79). The effects on CIMT were assessed in relationship to LDL-C and CRP levels obtained after 12 months of therapy. RESULTS: Changes in CIMT were directly related to the final LDL-C level obtained on statin therapy after 12 months (R = 0.219, p = 0.015). Carotid intima-media thickness regression was seen in 61% of the subjects in the lowest quartile of final LDL-C (< 70 mg/dl) versus 29% of the subjects with the highest quartile of final LDL-C (> or = 114 mg/dl, p = 0.008). No threshold value was seen, with more favorable effects on absolute change in CIMT with lower values of LDL-C (decrease in CIMT of 0.06 +/- 0.17 mm in the lowest quartile compared with an increase of 0.06 +/- 0.09 in the highest quartile of LDL-C, p = 0.008). On-treatment LDL and CRP concentrations both below the group median values were associated with the greatest likelihood of CIMT regression. CONCLUSIONS: Regression of carotid atherosclerosis is directly related to the absolute LDL-C level on statin therapy. The greatest regression was obtained with an LDL-C < 70 mg/dl, supporting marked LDL-C reduction to levels below current NCEP guidelines. PMID- 14743851 TI - Endomyocardial fibrosis in Churg-Strauss syndrome. PMID- 14743852 TI - Need for hospice and palliative care services in patients with end-stage heart failure treated with intermittent infusion of inotropes. AB - BACKGROUND: Hospice and palliative care programs to relieve suffering and optimize management of terminally ill patients have grown rapidly in the United States. However, there are no data on the need for these services among patients with end-stage heart failure receiving intermittent infusion of intravenous inotropes. HYPOTHESIS: The need for hospice and palliative care programs among patients in end-stage heart failure who receive intermittent infusion of inotropes is investigated. METHODS: The study included all stable patients with refractory heart failure symptoms treated with inotropes in our outpatient unit. A total of 73 patients (65 +/- 12 years; left ventricular ejection fraction 22 +/ 9%; New York Heart Association class 3.6 +/- 0.4) were seen during a 49-month period. Of these, 35 patients (48%) met hospice or palliative care evaluation criteria upon referral but were offered, and accepted, the alternative of parenteral inotropes. In all, 1,737 individual outpatient treatment sessions were given, with a mean of 24 +/- 19 sessions per patient (range 5 to 118 sessions), representing a minimum of 9,948 h of inotrope therapy. RESULTS: A total of 18 (25%) patients died, 6 (8%) patients were withdrawn from the program (3 by their primary physicians and 3 because of significant travel limitations); 4 (5%) patients required continuous intravenous home therapy; and 44 (61%) patients were discharged with significant improvement in their heart failure symptoms. Only 7 of the 18 patients who died had received hospice or palliative care intervention, mainly for the sake of comfort and to ease the transition among family members. The rest of the patients were comfortable and had accepted the natural evolution of their disease; they were not interested in or did not require hospice or palliative care intervention. Of the patients discharged from the outpatient cardiac infusion unit, the interval free of heart failure symptoms after the final infusion treatment ranged from 201 to 489 days, with no need for hospitalization or emergency room visits. CONCLUSION: Our results demonstrate that intermittent infusion of intravenous inotropes can be safely administered and can improve symptoms in a significant number of patients, probably by slowing the natural progression of heart failure. Although the full clinical impact of inotrope therapy in an outpatient setting has not been fully defined, other nonhemodynamic-related benefits should be sought and investigated. Our results suggest that intermittent infusion of intravenous inotropes is one of the prominent variables that requires particular attention. In our experience, the institution of intermittent infusions of intravenous inotropes can, in fact, modify end-stage heart failure symptoms that, in most patients, are currently perceived to lead to a terminal event. Thus, appropriate use of intermittent infusion of intravenous inotropes may not only improve functional class and symptoms in a significant number of patients identified as terminal by their poor response to conventional therapy, but it may also facilitate better utilization of hospice and palliative care resources among patients with end-stage heart failure. Furthermore, the need for hospice and palliative care in patients with heart failure should be revisited in view of adjuvant treatment options such as intermittent infusion of intravenous inotropes. PMID- 14743853 TI - Postsystolic thickening detected by Doppler myocardial imaging: a marker of viability or ischemia in patients with myocardial infarction. AB - BACKGROUND: Postsystolic thickening (PST) of ischemic myocardial segments has been reported to account for the characteristic heterogeneity or regional asynchrony of myocardial wall motion during acute ischemia. HYPOTHESIS: Postsystolic thickening detected by Doppler myocardial imaging (DMI) could be a useful clinical index of myocardial viability or peri-infarction viability in patients with myocardial infarction (MI). METHODS: Doppler myocardial imaging was recorded at each stage of a standard dobutamine stress echocardiogram (DSE) in 20 patients (16 male, 60 +/- 13 years) with an MI in the territory of the left anterior descending artery. Myocardial velocity data were measured in the interventricular septum and apical inferior segment of the MI territory. Postsystolic thickening was identified if the absolute velocity of PST was higher than peak systolic velocity in the presence of either a resting PST > 2.0 cm/s or if PST doubled at low-dose dobutamine infusion. RESULTS: Doppler myocardial imaging data could be analyzed in 38 ischemic segments (95%), and PST was observed in 21 segments (55%), including 3 segments showing PST only at low-dose dobutamine infusion. There was no significant difference of baseline wall motion score index (2.1 +/- 0.3 vs. 2.1 +/- 0.6, p = 0.77) or peak systolic velocity (1.1 +/- 1.1 vs. 1.9 +/- 2.0 cm/s, p = 0.05) between segments with and without PST. Peri-infarction ischemia or viability during DSE was more frequently observed in segments with PST than in those without (86 vs. 24%, p < 0.05). The sensitivity and specificity of PST for prediction of peri-infarction viability or ischemia was 82 and 81%, respectively. CONCLUSIONS: Postsystolic thickening in the infarct territory detected by DMI is closely related with peri-infarction ischemia or viability at DSE. PMID- 14743854 TI - Potential application of tissue Doppler imaging to assess regional left ventricular diastolic function in patients with hypertrophic cardiomyopathy: comparison with 123I-beta-methyl iodophenyl pentadecanoic acid myocardial scintigraphy. AB - BACKGROUND: Tissue Doppler imaging (TDI) has been utilized to evaluate left ventricular myocardial dysfunction in patients with hypertrophic cardiomyopathy (HCM); however, no clear explanation for the abnormality of TDI variables has been forthcoming. HYPOTHESIS: Peak negative myocardial velocity gradient (MVG) derived from TDI may correlate with a disorder of fatty acid metabolism in patients with HCM. METHODS: Tissue Doppler imaging and 123I-beta-methyl iodophenyl pentadecanoic acid (123I-BMIPP) myocardial scintigraphy were performed in 15 patients with asymmetric septal hypertrophy (mean age 47 +/- 18 years) and in 12 healthy controls (mean age 43 +/- 10 years). RESULTS: In early 123I-BMIPP images, accumulation defects were observed in the ventricular septum in 12 patients and in the posterior wall in 8 patients with HCM. Peak negative MVG in the ventricular septum (1.1 +/- 0.5 vs. 2.8 +/- 0.5, p < 0.0001) and posterior wall (5.2 +/- 1.4 vs. 6.7 +/- 0.8, p < 0.01 ) was significantly lower in the HCM group than in the controls; also, these parameters were significantly lower in patients with than in those without a defect in the region in question. The peak negative MVG in the ventricular septum and posterior wall correlated inversely with the washout rate in all subjects. CONCLUSIONS: Peak negative MVG according to TDI is related to disorder of fatty acid metabolism in the regional left ventricular myocardium of patients with HCM. PMID- 14743855 TI - Two large thrombi in the left atrium in an elderly patient with mitral valve stenosis. PMID- 14743856 TI - Mortality and rate of stroke or embolism in atrial fibrillation during long-term follow-up in the embolism in left atrial thrombi (ELAT) study. AB - BACKGROUND: Patients with atrial fibrillation (AF) have a higher mortality and risk of stroke/embolism than patients with sinus rhythm. HYPOTHESIS: The aim of the study was to assess the association of clinical and echocardiographic characteristics with mortality and stroke/embolism and the use of antithrombotic medication in the year 2000 in patients who participated 1990-1995 in the Embolism in Left Atrial Thrombi (ELAT) study. METHODS: The study included 409 outpatients with nonrheumatic AF (62 +/- 12 years, 36% women, 39% intermittent AF). Patients with thrombi received anticoagulation, patients without thrombi aspirin until follow-up in 1995; thereafter, anticoagulation according to clinical risk factors was recommended. Primary events were death and secondary events were stroke/embolism. All patients were contacted during the year 2000. RESULTS: Mean follow-up was 102 months. Mortality was 4%/year; the cause of death was cardiac (n = 84), fatal stroke (n = 26), malignancy (n = 23), sepsis (n = 5), and unknown (n = 24). Multivariate analysis identified age (p < 0.0001), heart failure (p = 0.0013), and reduced left ventricular systolic function (p = 0.0353) as predictors of mortality. Stroke/embolism occurred in 83 patients, with a rate of 3%/year. Multivariate analysis identified age (p = 0.0006) and previous stroke (p = 0.0454) as predictors of stroke/embolism. In the year 2000, 51 (21%) of the 247 surviving patients received no antithrombotic medication, 88 received (36%) oral anticoagulants, 102 (41%) acetylsalicylic acid, and 6 (2%) low-molecular heparin. CONCLUSIONS: Therapy for heart failure and oral anticoagulation in AF should be seriously considered, especially in elderly patients and in those with previous stroke. PMID- 14743857 TI - The imaging spectrum of Apical Hypertrophic Cardiomyopathy. PMID- 14743858 TI - Effect of Bachmann's bundle pacing on atrial fibrillation: electrophysiologic assessment. AB - BACKGROUND: It has recently been reported that simultaneous multisite atrial pacing, Bachmann's bundle (BB) pacing, and coronary sinus (CS) pacing are useful for preventing the induction of atrial fibrillation (AF). HYPOTHESIS: We investigated whether a simple pacing approach via BB could reduce the induction of AF by extrastimuli (S2) from the right atrial appendage (RAA). METHODS: Programmed electrical stimulation was performed from the RAA and the area of BB at the superior aspect of the atrial septum, and bipolar recordings were obtained from the RAA, BB, and CS in 14 patients. RESULTS: In five patients, AF was induced with critically timed RAA-S2 delivered during RAA pacing. However, AF was not induced in any patient when RAA-S2 was delivered during BB pacing. The duration of the P wave during BB pacing was significantly shorter than that during RAA pacing and sinus rhythm (BB 80 +/- 16 ms vs. RAA 106 +/- 36 ms vs. sinus rhythm 100 +/- 24 ms, p < 0.05). The intra-atrial conduction time to the distal coronary sinus (CSd) caused by early S2 at the RAA was significantly reduced by BB pacing (BB 114 +/- 22 ms vs. RAA 157 +/- 35 ms, p < 0.001). CONCLUSION: Bachmann's bundle pacing reduces atrial conduction time caused by RAA S2 and may be useful for preventing the induction of AF. PMID- 14743859 TI - Geoffrey O. Hartzler. PMID- 14743860 TI - How do UV photomorphogenic responses confer water stress tolerance? AB - Although ultraviolet-B (UV-B) radiation is potentially harmful, it is an important component of terrestrial radiation to which plants have been exposed since invading land. Since then, plants have evolved mechanisms to avoid and repair UV radiation damage; therefore, it is not surprising that photomorphogenic responses to UV-B are often assumed to be adaptations to harmful radiation. This presupposes that the function of the observed responses is to prevent UV damage. It has been hypothesized that, as with blue light, UV-B provides a signal important for normal plant development and might be perceived within developing plants through nondestructive processes, perhaps through UV-specific signal perception mechanisms. UV signal perception can lead to photomorphogenic responses that may confer adaptive advantages under conditions associated with high-light environments, such as water stress. Plant responses to UV radiation in this regard include changes in leaf area, leaf thickness, stomatal density, photosynthetic pigment production and altered stem elongation and branching patterns. Such responses may lead to altered transpiration rates and water-use efficiencies. For example, we found that the cumulative effect of ambient UV-B radiation upon stomatal density and conductance can lead to altered water-use efficiencies. In field settings, UV might more properly be viewed as a photomorphogenic signal than as a stressor. Hence, it might be insufficient to attempt to fully evaluate the adaptive roles of plant responses to UV-B cues upon stress tolerance by the simultaneous application of UV and drought stress during development. We propose that rather than examining a plant's response to combinations of stressors one might also examine how a plant's response to UV induces tolerance to subsequently applied stresses. PMID- 14743861 TI - Joint effects of elevated levels of ultraviolet-B radiation, carbon dioxide and ozone on plants. AB - There is growing interest regarding the joint effects of elevated levels of surface ultraviolet B (UV-B) radiation, carbon dioxide (CO2) and ozone (O3) on plants. Our current knowledge of this subject is too limited to draw any specific conclusions, although one might state that such effects are likely to be highly species dependent and may be more than additive, additive or less than additive. There are a number of uncertainties associated with the experimental protocols used and the conclusions reached in many studies. Nevertheless, in North America, there appear to be genotypes of three monocot crop species (Avena sativa L., Oryza sativa L. and Sorghum vulgare L.); six dicot crops (Cucumis sativus L., Lactuca sativa L., Lycopersicon esculentum Mill., Phaseolus vulgaris L., Pisum sativum L. and Solanum tuberosum L.) and two conifer species (Pinus ponderosa Dougl. and Pinus taeda L.) that may be considered sensitive to the joint effects of elevated levels of UV-B, CO2 and O3. However, to provide a more reliable assessment or validation of the predictions, future research must consider the concept of plant response surfaces and describe them more fully in numerical terms. Achieving that objective will require close cooperation among a number of scientists representing geographic locations with known spatial and temporal differences in UV-B, CO2 and O3 to conduct experiments under their site-specific conditions, using common plant materials and experimental protocols. PMID- 14743862 TI - Protochlorophyllide reduction: mechanisms and evolutions. AB - Protochlorophyllide (Pchlide) reductases are key enzymes in the process of chlorophyll biosynthesis. In this review, current knowledge on the molecular organization, substrate specificity and assembly of the light-dependent reduced nicotinamide adenine dinucleotide phosphate:Pchlide oxidoreductases are discussed. Characteristics of light-independent enzymes are also described briefly, and the possible reasons for the selection of light-dependent enzymes during the course of evolution are discussed. PMID- 14743863 TI - Comprehensive photokinetic and NMR study of a biphotochromic supermolecule involving two naphthopyrans linked to a central thiophene unit through acetylenic bonds. AB - A photophysical and photochemical study of a biphotochromic compound where two naphthopyran units are linked by an acetylene-thiophene-acetylene bridge has been carried out in toluene. Both fluorescence and intersystem crossing to the triplet manifold were found to compete with the photocoloration process. Two photoproducts (transoid-trans and transoid-cis stereoisomers), absorbing at approximately 480 nm and corresponding to the opening of a single photochromic unit, were detected by spectrophotometric analysis after short irradiation time in diluted solution and identified by 1H-nuclear magnetic resonance (NMR) spectroscopy. After prolonged irradiation at 228 K of highly concentrated solutions (up to 3 x 10(-3) mol dm(-3)), two additional isomers, absorbing at approximately 550 nm, were formed. Their NMR spectra indicate the opening of both photochromic units. An interesting effect of selective vibronic excitation was found, showing that the photoreaction is favored at excited vibronic levels to the detriment of the radiative relaxation. PMID- 14743864 TI - Reactive oxygen species formation by UV-A irradiation of urocanic acid and the role of trace metals in this chemistry. AB - We have extended our study of the decomposition of urocanic acid (UCA) with ultraviolet A radiation (UV-A) by the self-sensitized generation of singlet oxygen (see Photochem. Photobiol. 75, 565 [2002]). The chemistry has been found to be partially dependent on the presence of trace metal, most likely iron. Rigorous removal of metal impurities from the reaction mixture, using Chelex, retarded (but did not eliminate) the UV-A-initiated UCA degradation. The addition of small amounts of ferric chloride to the Chelex-treated solutions restored reactivity. Chelex treatment had a modest effect on the previously reported ability of UCA photoproducts to photonick supercoiled plasmid DNA. Also, photoinactivation of Sindbis virus on irradiation with the UCA photoproducts is now reported. Inactivation of the virus by a photoproduct mixture derived from a UCA solution that had been pretreated with Chelex was less rapid and gave better behaved time-course plots than was observed for photoproducts from non-Chelex treated solutions. These results are particularly noteworthy in light of the ubiquitous presence of both UCA and iron in the skin. PMID- 14743865 TI - Single-molecule fluorescence spectroscopy of TOTO on poly-AT and poly-GC DNA. AB - Excited state lifetime and amplitude measurements were made on thiazole orange dimer (TOTO), a dimeric DNA-intercalating fluorophore, at single-molecule concentrations. As expected from previous study, the excited state lifetime of TOTO intercalated in DNA is dependent on the sequence of the double-stranded DNA, having values of 2.2 ns in poly-GC DNA and 1.8 ns in poly-AT DNA. The distribution of excited state lifetimes of single molecules of TOTO intercalated into oligonucleotides having varying proportions of poly-GC sequences relative to poly-AT sequences were analyzed as a function of the fraction of poly-GC. By using excited state lifetime distributions from the purely GC and purely AT oligonucleotides as a basis set, it was possible to estimate the GC content of oligonucleotides with intermediate GC composition to within a few percent error. This serves as a model for the analysis of equilibrium binding distributions in DNA using single-molecule methods. PMID- 14743866 TI - TOTO binding affinity analysis using single-molecule fluorescence spectroscopy. AB - The sequence dependence of the double-stranded DNA (dsDNA)-binding affinity of TOTO, a thiazole orange dimer that functions as a DNA-intercalating fluorophore, was measured using single-molecule methods. An analysis was performed of the distribution of excited-state lifetimes of single molecules of TOTO intercalated into dsDNA fragments containing four-base pair sequences shown previously to have high affinity for TOTO under conditions used in nuclear magnetic resonance (NMR) spectroscopy. For the current studies, the putative binding sites were located centrally in 30-base pair-long dsDNA fragments in which the remaining sequence was either all poly-AT or poly-GC. The lifetime of TOTO fluorescence when bound to these fragments was entirely determined by the background sequence, i.e. DNA fragments with a poly-AT background predominantly gave a fluorescence lifetime of 1.7 ns, whereas DNA fragments with a poly-GC background gave a lifetime of 2.0 ns, independent of the presence or absence of the putative binding sequence. By performing competitive binding experiments in which these DNA fragments competed for TOTO binding with pure poly-AT fragments and using single-molecule fluorescence methods to determine the number of each type of DNA fragment having a TOTO bound in an equilibrium mixture, the relative binding affinity of each putative binding site was determined. The results of these experiments showed clearly that TOTO has no preference for binding to the putative binding sites over binding poly-AT or poly-GC under the conditions of these measurements. This suggests that there is very little sequence dependence of TOTO binding under conditions that would likely predominate in most biological applications of this intercalating dye. PMID- 14743867 TI - The embedding of meta-tetra(hydroxyphenyl)-chlorin into silica nanoparticle platforms for photodynamic therapy and their singlet oxygen production and pH dependent optical properties. AB - This study relates to nanoparticle (NP) platforms that attach to tumor cells externally and only deliver singlet oxygen for photodynamic therapy (PDT) while conserving the embedded photosensitizers (PS). As a model, we demonstrate the successful embedding of the PS meta-tetra(hydroxyphenyl)-chlorin (m-THPC) in NP that are based on a sol-gel silica matrix and also show its positive effect on the singlet oxygen production. The embedding of m-THPC inside silica NP is accomplished by a modified Stober sol-gel process, in which (3-aminopropyl) triethoxysilane is introduced during the reaction. Singlet oxygen delivery by the targetable photodynamic NP exceeds that from free PS molecules. In the physiological pH range, there is no significant pH-induced decrease in the fluorescence of m-THPC embedded in silica NP, which might otherwise affect the efficiency of PDT. PMID- 14743868 TI - Visible light irradiation of ethidium bromide-stained interphase nuclei causes DNA-protein linking and structural stabilization of nucleoprotein complexes. AB - Fixation of DNA and proteins in the isolated rat hepatocyte nuclei stained with ethidium bromide and irradiated with visible light was analyzed in this study. It was shown that irradiation results in the following modifications of higher-level nucleoprotein complexes of interphase chromatin: (1) the complexes acquire resistance to decondensing treatments, which may be indicative of the formation of links between proteins or proteins and DNA in the chromatin; (2) the linking rate for both DNA and proteins is dose dependent; (3) the irradiation induces intermolecular link formation between DNA molecules, which brings about an increase in the average molecular weight of DNA fragments; (4) some modifications (dimerization, etc.) of histones and nonhistone proteins occur; and (5) histone proteins are not effectively cross-linked to DNA. The structural stabilization of interphase chromatin is possibly mediated by free radical-based mechanisms, whereas disulfide bonds seem to play no significant role in the cross-linking. PMID- 14743869 TI - Fluorescence fingerprints of Eisenia fetida and Eisenia andrei. AB - We describe a fluorescent method that allows to differentiate the worms Eisenia fetida and Eisenia andrei. In fact, the coelomic fluid of E. andrei displays specific fluorescence absent in that of E. fetida. The two species do not metabolize the same types of molecules and thus can be differentiated at the molecular level. Each species has specific fluorescence fingerprints. PMID- 14743870 TI - Simplified calibration for broadband solar ultraviolet radiation measurements. AB - Aspects of different calibration procedures for erythemally weighing broadband radiometers are presented in this study. These instruments are common in projects dealing with ultraviolet radiation effects on humans. Many erythemally weighing broadband radiometers are still operated using a single calibration factor (cf) that is provided with the instrument. The individual characteristics of every instrument are strongly dependent on the total ozone amount and the solar elevation. Therefore, a calibration procedure also has to take into account the ozone concentrations and the solar elevation to compensate for the effects of the individual characteristics and to provide comparable measurements. Given the variation of the ozone concentrations and the solar elevation, an individual cf has to be calculated for every measurement. Using a simplified version of the calibration procedure, which is presented in this study, can lessen this effort. Taking into account the relevant meteorological conditions for a measuring site, a single cf is calculated to compensate the individual characteristics of the instruments and therefore deliver comparable measurements with less effort. PMID- 14743871 TI - Accumulation and photocytotoxicity of hypericin and analogs in two- and three dimensional cultures of transitional cell carcinoma cells. AB - The aim of this study was to investigate the in vitro cellular accumulation, distribution and photocytotoxic effect of hypericin in two-dimensional (2-D) and three-dimensional (3-D) cultured RT-112 transitional cell carcinoma cells of the bladder. In addition, two iodinated derivatives of hypericin were incorporated to investigate whether these analogs, with their increased lipophilicity and heavy atom effect, display a different biological behavior and optimized photodynamic effect. The results indicate that hypericin and mono-iodohypericin behave similarly in terms of cellular accumulation, spheroidal distribution and photocytotoxic effect. In contrast, di-iodohypericin concentrated to a higher extent in monolayers and spheroids, but the accumulation was restricted to the outermost part of the spheroid. An inverse correlation therefore seems to exist between the extent of cellular uptake under 2-D conditions and the penetration of the compounds in multicellular systems. Moreover, a less pronounced photocytotoxic effect was observed for di-iodohypericin in both 2-D and 3-D cell culture systems. It can be concluded that iodinated derivatives of hypericin do not show an increased cytotoxic effect upon irradiation in either monolayers or spheroids. Moreover, this study shows that when new photosensitizers are preclinically developed, the use of 3-D cell aggregates is critical for a correct evaluation of their efficacy. PMID- 14743873 TI - On the photobiological properties of chimeras combining quaternary ammonium derivatives of retinoic amides and psoralen. A study with cultured human keratinocytes. AB - The effectiveness of the combination of retinoids with 8-methoxypsoralen (8-MOP) and ultraviolet-A (UV-A) light in the treatment of some cutaneous proliferative diseases has motivated the synthesis of new "chimera-type" molecules built from psoralen derivatives and retinoic amides and related molecules. The chimeras result from the combination of 8-(3-bromopropyloxy)-psoralen with amides prepared by reacting 4-amino-pyridine with 13E- and 13Z-retinoic acids or a "retinoid like" derivative with an alkene chain of only three double bonds. The synthesis of chimeras built with the 8-(3-bromopropyloxy)-psoralen and the amide of cinnamic acid or its 4-methoxy derivative has also been carried out. In contrast to 8-MOP, all the chimeras exhibit strong molar absorptivities in the range 20 000-40 000 M(-1) cm(-1) in the 340-390 nm UV-A region. The "retinoid-like"- and retinoid-psoralen chimeras are characterized by a marked dark toxicity toward proliferating NCTC 2544 keratinocytes (with a lethal dose corresponding to 50% cell survival [LD50] of 1-5 microM) as compared with that of the cinnamic acid derivative-psoralen chimeras (LD50 > or = 50 microM). This toxicity leads to alteration of the mitochondrial membrane potential. At nontoxic concentrations, the chimeras demonstrate effective psoralens + UV-A-induced photocytotoxicity. They are moderate photosensitizers of membrane lipid peroxidation. Cell apoptosis is a major photocytotoxic process as suggested by the fluorescence-activated cell sorting technique using annexin-fluorescein isothiocyanate and propidium iodide as apoptotic markers. PMID- 14743872 TI - Activation of heat shock protein 70 promoter with meso-tetrahydroxyphenyl chlorin photodynamic therapy reported by green fluorescent protein in vitro and in vivo. AB - Cellular responses to photodynamic therapy (PDT) include induction of heat shock proteins (HSP). We examined meso-tetrahydroxyphenyl chlorin (mTHPC) PDT-mediated HSP activation in EMT6 cells stably transfected with a plasmid containing the gene for green fluorescent protein (GFP) driven by an hsp70 promoter. mTHPC incubation induced concentration-dependent GFP expression. Irradiation of cells exposed to a sensitizer concentration that induced a slight increase in GFP and no loss of cell viability resulted in fluence-dependent GFP accumulation. In response to drug only and to PDT, GFP levels increased to a maximum of four- to five-fold above control levels with increasing drug or fluence and then decreased at higher doses. A trypan blue-exclusion assay confirmed that decreased GFP levels in both cases were due to a loss of cell viability. For initial evaluation in vivo, HSP70/ GFP-transfected EMT6 tumors were grown in BALB/c mice and subjected to mTHPC-PDT with a fluence of 1 J/cm2. Six hours after PDT, GFP fluorescence was imaged in these tumors through the intact skin in vivo. These results indicate that sublethal doses of mTHPC-PDT stimulate GFP expression under the control of an hsp70 promoter and illustrate the potential of noninvasively monitoring reporter protein fluorescence as a measure of molecular response to PDT. PMID- 14743874 TI - Susceptibility to basal cell carcinoma is associated with high dermal mast cell prevalence in non-sun-exposed skin for an Australian populations. AB - In a Danish population, basal cell carcinoma (BCC) patients have a higher dermal mast cell prevalence in buttock skin than controls. This finding was supported by a functional link in mice between histamine-staining dermal mast cells and the extent of susceptibility to UV-B-induced systemic immunomodulation. It was important to confirm that this association was maintained in an Australian population with very different ancestry and sun exposure patterns. Australian BCC patients (n = 26) had significantly higher densities of mast cells in the dermis of buttock skin than control subjects (n = 25) (P = 0.0003, Mann-Whitney U-test). However, this correlation was lost at the sun-exposed site of the hand (P = 0.547, Mann-Whitney U-test). To further evaluate whether a relationship exists between dermal mast cell prevalence in sun-exposed skin and incidence of BCC in a larger study, biopsies of dorsal hand skin were obtained from an age-stratified random sample of 166 Queensland subjects, together with the 51 South Australian subjects, and dermal mast cell prevalence was quantified. Older subjects (over the median age of 42 years) had a greater incidence of BCC development (P = 0.0001, chi-square test) and significantly higher mast cell densities in hand skin (P = 0.0001, chi-square test) than younger subjects. However, mast cell density in sun-exposed hand skin was not significantly associated with BCC incidence. Finally, cellular expression of c-kit correlated with mast cell prevalence in non-sun-exposed skin, thereby implicating the stem cell factor-c kit axis in the intrinsic mechanisms that regulate prevalence. These results show that high prevalence of dermal mast cells in buttock skin but not hand is associated with BCC development in an Australian population. PMID- 14743875 TI - Simulation of human vision by image evolution in light-excitable nonlinear optical mediums. AB - The nonlinear optical membrane used for image processing was prepared using tetraethyl silicate by sol-gel method under mild conditions. Ru(bpy)3Cl2 (bpy, bipyridine) was immobilized in the sol-gel matrix. This light-excitable membrane containing Ru(bpy)3Cl2 as photocatalyst could catalyze the Belousov-Zhabotinsky reaction by illumination. Our results indicated that a fine-definition image processing could be autoperformed when projecting a picture on this medium. The imaging process was an image evolution process, which was similar to that of the human vision process. PMID- 14743876 TI - Whom and how to treat: weighing the costs and effects. AB - Taking the right decision in health care may be as much an art as a science. However, without exception and whatever the decision, usage is made of scarce resources and they may be used in an alternative way. This implies costs and there comes a time at which the various options have to be assessed in comparison with their effects. Cost-effectiveness aims at informing decision-makers about the balance between the costs and effects of alternative therapies. PMID- 14743877 TI - Sense and nonsense of lipids in artificial nutrition. AB - Lipids are a component of artificial nutrition. For 20 years lipids covered only a limited percentage of the energy requirement of the patient, depending on parenteral nutrition. Ideas about dose, type and aims of lipids administration in artificial nutrition have changed during this period. The study is based on the discussion around lipids in artificial nutrition. PMID- 14743878 TI - Probiotics in gastroenterology: indications and future perspectives. AB - Nowadays. there is a growing interest in probiotics as a safe way of changing the intestinal bacterial flora. Probiotics may have potential in several gastroenterological conditions, especially when the intestinal flora has been disturbed. Most scientific evidence is available for diarrhoea patients treated with Lactobacillus GG, Lactobacillus reuteri or Saccharomyces boulardii. Meta analyses have shown an overall reduction in the risk of antibiotic-associated diarrhoea during treatment with probiotics, and benefits have also been demonstrated for patients with rota-virus-associated diarrhoea. Patients with inflammatory bowel disease, an inflammatory disorder characterized by a change in the intestinal flora, are another important target group for which probiotics may be beneficial. It has been claimed that in ulcerative colitis and Crohn disease patients, lactobacilli, S. boulardii and Escherichia coli reduce relapses. but most studies were not placebo-controlled. A reduction in relapses has also been demonstrated in pouchitis patients treated with a multispecies probiotic. Irritable bowel syndrome might be another clinical indication for probiotic therapy, but results of clinical trials performed in these patients are inconsistent. Additionally, probiotics may improve lactose absorption. Helicobacter pylori eradication and constipation. Finally, in animal models of colorectal cancer, treatment with probiotics reduces the prevalence of this disease, and in humans the amount of genotoxic substances in faeces has been reduced. In conclusion, the results of studies on the effects of probiotics in gastrointestinal conditions are encouraging. but well-designed placebo-controlled studies are warranted before recommendations for therapeutic or preventive use can be given. Many issues still have to be resolved, including optimal dose and duration of treatment, selection of and differences between the several available probiotic strains, and, importantly, their mechanisms of actions have to be elucidated. PMID- 14743879 TI - Clinical application of video capsule endoscopy. AB - BACKGROUND: Non-invasive imaging of the complete small bowel has become possible with the recently available video capsule endoscopy technique. The aim of this study was to review and assess the possibilities and limitations of the clinical application of video capsule endoscopy. METHODS: Literature review. RESULTS: Video capsule endoscopy is a new promising technique with good to excellent visualization of the mucosa of the small bowel. It has a high diagnostic yield in patients with occult gastrointestinal blood loss (OGIB) and Crohn disease (CD). Several other indications need further study. Compared to small-bowel X-ray and push enteroscopy, diagnostic yield appears to be superior. Video capsule endoscopy is very well tolerated by patients. Its most important limitation is the impossibility of taking biopsies and therapeutic interventions. CONCLUSION: Video capsule endoscopy is a promising new diagnostic tool for small-bowel disease. PMID- 14743880 TI - Endoscopic treatment of gastro-oesophageal reflux disease: the new kid on the block. AB - BACKGROUND: Gastro-oesophageal reflux disease is a common disease entity with approximately 7% of European adults experiencing significant daily symptoms. The impact of reflux disease on the quality of life is considerable. Complications of reflux disease include oesophagitis, stricture, Barrett's and pulmonary symptoms. Most patients can be adequately managed by treatment with a proton-pump inhibitor. However, symptom relapse is common after cessation of therapy, thus many patients are committed to life-long therapy. Until recently, anti-reflux surgery was the single therapeutic alternative. Now, novel endoscopic techniques have become available to treat patients suffering from reflux disease. Application of these techniques is challenging. METHODS: Update on new endoscopic techniques for treatment of reflux discase. RESULTS: Currently available endoscopic techniques include endoscopic suturing, radiofrequency ablation and biopolymer injection. Interventions typically take 30-40 min and can be performed under conscious sedation. First reports describe successful reduction of symptoms. Six months after therapy. reportedly 58%-85% of patients are off proton pump inhibition. Yet, there are conflicting results on 24-h pH measurement and insufficient data on the mechanism of altered oesophageal motility. Long-term data are not yet available. In our series of over 50 procedures, no serious complications have occurred. CONCLUSIONS: Endoscopic treatment of reflux disease is feasible and safe. Techniques reduce both symptoms and medication use associated with the disease, albeit with an uncertain long-term outcome. As pursuit of this technology is appealing, techniques are being introduced before thorough comparison and evaluation of therapeutic benefit have been completed. Comparative studies between conventional anti-reflux treatment and various luminal anti-reflux therapies are needed and long-term efficacy remains to be established. PMID- 14743881 TI - Transanal endoscopic microsurgery. AB - BACKGROUND: Transanal endoscopic microsurgery (TEM) is a newly developed, minimally invasive technique for the local resection of rectal tumours. Its place needs to be defined. METHODS: Literature review. RESULTS: Local excision of rectal tumours is associated with low mortality and morbidity. It is indicated in adenomas for curation and in rectal cancer for palliation. Transanal resection is used most frequently, but its use is limited. Other local techniques have no added value and are used sparingly. TEM is a newly developed, minimally invasive technique. Its technical characteristics allow an excellent view also in larger and more proximal tumours. Compared to other local techniques, mortality and morbidity are minimal and a laparotomy is more often preventable. Proper histological examination is possible and free margins are almost always observed. In adenomas it results in hardly any recurrences and in rectal cancer it can be adequately used for palliation. The results following local resection for rectal cancer with curative intent are promising, but they are only described anecdotally. CONCLUSIONS: TEM is an elegant technique with excellent results. It imposes itself as a method of choice for the local resection of rectal adenomas and of rectal cancer for palliation. If curation is intended in rectal carcinomas, TEM should be exercised with caution. Proper judgement over existing local techniques and TEM is being impeded by a lack of scientific argumentation. PMID- 14743882 TI - Pathophysiological characteristics of long- and short-segment Barrett's oesophagus. AB - BACKGROUND: Depending on the length of the segment of columnar epithelium in the distal oesophagus, Barrett's oesophagus can be divided into long-segment and short-segment Barrett's oesophagus. This article describes the pathophysiological characteristics of both forms of Barrett's oesophagus. METHODS: Review of the literature. RESULTS: Although there is some disagreement, ours and most other studies suggest that long-segment columnar-lined oesophagus is causally linked to chronic gastro-oesophageal reflux and that this from of Barrett's oesophagus is characterized by lower oesophageal sphincter tone, reduced oesophageal contractility and increased acid reflux. Short-segment Barrett's oesophagus is also associated with acid reflux, but the degree of oesophageal acid exposure and the level of other pathophysiological alterations seem to be lower. CONCLUSION: Pathophysiological abnormalities appear to be more prominent in long-segment Barrett's oesophagus than in short-segment Barrett's oesophagus. PMID- 14743883 TI - Dyspepsia management in primary care. AB - BACKGROUND: Dyspepsia is common in western society. Prompt endoscopy is imperative in all patients with sinister symptoms or if symptoms first appear after the age of 50-55 years, but the optimal management of younger patients with uncomplicated dyspepsia is still open to debate. METHODS: The literature on the management of uncomplicated dyspepsia is reviewed and a personal view is presented. RESULTS: Strategies based on non-invasive detection of Helicobacter pylori are probably the most cost-effective. Currently (H. pylori prevalence 30% 40%), a 'test and treat' approach using a non-invasive test to detect H. pylori is likely to be the most efficient first step. If the patient is H. pylori negative or if symptoms persist after successful H. pylori eradication, empirical treatment with an anti-secretory drug is justified. Endoscopy is reserved for those patients in whom this approach fails. If the prevalence of H. pylori decreases, the positive predictive value of any non-invasive H. pylori test will become too low. A 'test and scope' approach in which a positive test can be confirmed by two or more biopsy-based tests is then more appropriate. At a very low prevalence of H. pylori in the dyspeptic population, non-invasive testing for H. pylori loses its significance and empirical treatment with an antisecretory drug becomes a rational first step. CONCLUSIONS: The optimal approach to management of uncomplicated dyspepsia is dependent on the prevalence of H. pylori among the dyspeptic population. At the current prevalence rate, 'test and treat', followed by acid suppressive treatment in the case of persisting symptoms, is the most appropriate strategy. PMID- 14743884 TI - Haemostasis in inflammatory bowel diseases: clinical relevance. AB - BACKGROUND: In inflammatory bowel disease (IBD), many alterations of haemostasis have been reported. Furthermore. IBD is associated with thromboembolic disease. METHODS: Literature on new insights into the physiology of haemostasis. the interaction between haemostasis and inflammation, plasmatic and mucosal changes in haemostasis in IBD, and haemostasis-interfering therapy in patients with IBD are reviewed. RESULTS: Haemostasis is a vascular-bed-specific, locally regulated, physiological phenomenon aimed at the maintenance of fluidity of blood, and not a simple cascade-shaped chain of reactions. Coagulation activation is part of the inflammatory response, but coagulation activation induces pro-inflammatory effects at the same time. Coagulation and fibrinolysis are activated in the course of IBD. but sometimes in a misbalanced way. Overall, this may induce a state of plasmatic hypercoagulation, irrespective of disease activity. Thromboembolic disease is a common extra-intestinal manifestation of IBD. Established treatment of thromboembolism is similarly useful in IBD patients: concurrent aggressive treatment of exacerbations is recommended. Intractable gastrointestinal bleeding seldom occurs, and has mainly been reported in patients with untreated active IBD. Thrombophilic genetic background does not seem to be responsible for either the increased risk of thromboembolism, or the prevalence of IBD. Haemostasis-interfering therapy to alter the course of IBD is experimental. CONCLUSION: Thromboembolism is an extraintestinal manifestation of IBD, partly because of the associated state of plasmatic hypercoagulation. Thrombophilic genetic background does not contribute to prevalence or course of IBD: genetic investigations may be restricted to patients with clinically proven thrombophilia. Anticoagulant therapy can normally be given to patients. but not as an established therapy against IBD. PMID- 14743885 TI - IBD and genetics: new developments. AB - BACKGROUND: Inflammatory bowel disease (IBD) is a complex disorder with an aetiology that is only partly understood. Apart from environmental factors, inheritance contributes to IBD. REVIEW: Family studies show an increased risk among family members of a patient with IBD, particularly among first-degree relatives. In twin studies, concordance for disease type and localization is observed. In genetically isolated groups there is a higher prevalence of IBD. For instance. Ashkenazi Jews carry the highest risk. Further evidence comes from animal species that spontaneously develop IBD. Unlike Mendelian inheritance, in complex genetic diseases like IBD, genes are expected to be low penetrant and therefore less prone to selection, which results in higher expected gene frequencies. NOD2/CARD15, the first gene associated with IBD, is a polymorphic gene involved in the innate immune system. The gene has over 60 variations. Three of these play a role in 27% of patients with CD, with a predilection for patients with ileal disease. CONCLUSION: Genetics plays an important role in unravelling the pathogenesis of IBD leading to possible new therapeutic approaches. PMID- 14743886 TI - Safety of thiopurines in the treatment of inflammatory bowel disease. AB - BACKGROUND: Thiopurines have proven efficacy in inflammatory bowel disease. However, concerns regarding toxicity have limited the use of these agents as first line of medical therapy. METHODS: Review of the literature regarding metabolism, efficacy and side effects. RESULTS: In clinical trials, up to 15% of patients discontinued 6-mercaptopurine or its pro-drug azathioprine prematurely due to adverse events. These events may be divided into dose-independent idiosyncratic reactions and dose-related, pharmacologically explainable toxicity. Dose-independent reactions include skin rash, fever, diarrhoea and pancreatitis. Most frequently observed dose-dependent adverse events are nausea, malaise and myelotoxicity. Furthermore, dose-dependent and dose-independent hepatotoxicity may occur. Recent insights obtained by therapeutic drug monitoring in patients on azathioprine or 6-mercaptopurine have led to strategies to reduce toxicity. One strategy is to detect poor metabolisers of thiopurines by establishing the activity of the key enzyme thiopurine methyltransferase. However, the clinical relevance of this strategy is still a point of debate. Another strategy is to administer 6-thioguanine, which is an agent close to the effective 6-thioguanine nucleotides. CONCLUSION: Therapeutic drug monitoring of thiopurines resulted in strategies to reduce toxicity. The value of these strategies has yet to be proven in prospective randomized trials. PMID- 14743887 TI - Screening for colorectal cancer: medical and economic aspects. AB - BACKGROUND: Colorectal cancer (CRC) is the second commonest cause of cancer death in the Western world. In The Netherlands, CRC causes about 4400 deaths per year, and its diagnosis and treatment make up for a large share of health-care costs. METHODS: Review and discussioN. RESULTS: Experts in the field presently assume that screening for CRC and its precursor lesions, colorectal adenomas (CRAs), could prevent death from colorectal neoplasia by more than 80%. Additionally, there is increasing acknowledgement that CRC screening programmes can save lives at a cost similar to, or even less than, the generally accepted breast cancer or cervical cancer screening programmes. Nonetheless, while neighbouring countries have taken vigorous measures to fight CRC, the Dutch are still hesitating in this matter. This is partly due to some yet unanswered questions concerning the acceptability of screening for CRC in the general population, the starting age and the frequency of screening, the type of screening tests to be used, and the programme organization. In this commentary, general epidemiological and pathogenetic aspects of CRC are addressed. In addition, some frequently asked questions (FAQ) and (very subjective) answers about screening for CRC are offered, as potential substrate for further in-depth discussions. CONCLUSION: The emerging message for the community is that an effective national screening programme is urgently required to reduce the substantial morbidity and mortality from this disease. PMID- 14743888 TI - New developments in systemic chemotherapy in advanced colorectal cancer. AB - BACKGROUND: The majority of patients with newly diagnosed colorectal cancer who present with concurrent metastases are considered to be incurable from the disease. For their treatment, these patients depend on systemic anticancer therapy and supportive care. METHODS: This article reviews recent developments in systemic treatment of disseminated colorectal cancer. RESULTS: Until recently, the fluoropyrimidines were the only cytotoxic drugs available, but during recent years three other classes of active conventional cytotoxic drugs and a new class of target-directed drugs have become available. New cytotoxic drug therapies include raltitrexed, irinotecan and oxaliplatin. Patients receiving irinotecan experience a better quality of life than those treated with best supportive care alone. Irinotecan or oxaliplatin in combination with 5-fluoro-uracil (5-FU) and leucovorin are currently the most active treatment regimens. Oral prodrugs of 5 FU, such as capecitabine and uracil, have been developed in order to mimic the protracted infusion schedule of 5-FU, and these drugs may change the daily practice of palliative chemotherapy for colorectal cancer in the coming years. Therapeutic agents that target specific molecular processes that promote proliferation, vascularization and metastasis, and inhibit apoptosis, are being designed and may offer a rational approach to anticancer therapy. Examples of this novel approach are monoclonal antibodies and small molecules adhering to the epithelial growth factor receptor and vascular endothelial growth factor receptor in order to inhibit growth stimulation and angiogenesis. CONCLUSION: The current debate is no longer whether to use palliative chemotherapy in metastatic colorectal, but which patient will benefit from which combination and in what sequence. PMID- 14743889 TI - Role of cyclooxygenase-2 in the development and treatment of oesophageal adenocarcinoma. AB - BACKGROUND: Various studies suggest that aspirin and other non-steroidal anti inflammatory drugs (NSAIDs) are promising anticancer agents. Epidemiological studies have found that long-term use of NSAIDs is associated with a reduced incidence of colorectal, gastric and oesophageal cancers, while experimental and clinical studies have demonstrated that treatment with NSAIDs causes a statistically significant reduction in both the number and the size of polyps in familial adenomatous polyposis (FAP) patients. METHODS: In this review, the mechanisms by which NSAIDs exert their chemopreventive and antineoplastic effects are described. RESULTS: Although the precise anticancer actions of NSAIDs are not fully explained, they probably involve inhibition of cyclooxygenase (COX), which is the rate-limiting enzyme in the conversion of arachidonic acid to prostaglandins. Two isoforms of this enzyme (COX-1 and COX-2) have been identified. COX-1 is constitutively expressed and considered to be a housekeeping gene, while COX-2 is not usually detectable in normal tissues, but can be readily induced in processes like inflammation, reproduction and carcinogenesis. The mechanisms by which COX-2 is thought to be involved in the carcinogenesis include resisting apoptosis, increasing cell proliferation, stimulating angiogenesis and modulating the invasive properties of cancer cells. CONCLUSION: This report reviews the mechanisms by which COX-2 can contribute to carcinogenesis, its role in prognosis, and the possible place of selective COX-2 inhibitors in the prevention and treatment of gastrointestinal malignancies, focusing particularly on oesophageal cancer. PMID- 14743890 TI - Surveillance for familial pancreatic cancer. AB - BACKGROUND: It is estimated that between 5% and 10% of pancreatic cancer (PC) cases are due to hereditary factors. METHODS: Review of the literature. RESULTS: In families with clustering of PC, germline mutations in specific genes might be responsible for the disease. It is suggested that PC progresses from precursor lesions, the pancreatic intraepithelial neoplasias (PanINs). Several key genetic alterations in oncogenes (K-ras, Her2/neu) and tumour suppressor genes (p16, p53, SMAD4) occur in the progression from PanIN lesions towards PC. PC is mostly diagnosed on clinical presentation at an advanced, no longer resectable, stage. The overall 5-year survival rate is extremely poor. Recent studies report a better survival rate of PC, providing surgery takes place at an early stage. Surveillance of family members at increased risk for PC might lead to detection of tumours at an early stage and improve overall survival. CONCLUSION: Clinicians should be aware of the tumour syndromes that are associated with an increased risk of PC. Efforts to improve PC survival must focus on identification of high risk patients, detection of early stage disease and novel screening strategies. PMID- 14743891 TI - Transjugular intrahepatic portosystemic shunt (TIPS): indications and long-term patency. AB - BACKGROUND: Since the introduction of TIPS (the transjugular intrahepatic portosystemic shunt) into clinical practice in 1989, substantial knowledge, partially derived from controlled trials, has become available regarding technical and clinical aspects of the procedure. A number of prospective studies have assessed the long-term patency of radiological shunts, the recognized main technical weakness of the procedure. METHODS: Review of published data regarding the optimal indications and long-term patency of TIPS. RESULTS: Information on the long-term patency of TIPS is surprisingly scarce. Within 2 years of TIPS creation, re-interventions to re-establish or maintain the patency of the shunt are required in 70%-90% of patients, and in 20%-40% total occlusion develops. Limited available data suggest, however, that in about 80% of patients the shunt is patent after 3-5 years. There is consensus that TIPS is a main, second-line treatment option for variceal haemorrhage not responding to other therapies. Although widely used for treating refractory ascites, gastric variceal bleeding and Budd-Chiari syndrome, these indications require more study. A number of other potential indications remain poorly defined. CONCLUSION: TIPS is a major treatment modality to manage the complications of portal hypertension and Budd Chiari syndrome. The available data indicate that TIPS is not only a short-term treatment option but may provide long-term portosystemic decompression. Technical improvements, e.g. the use of covered or drug-eluting stents, are essential to reduce the high rate of shunt dysfunction. PMID- 14743892 TI - Hepatitis C 2002 guidelines: summary and annotations. AB - BACKGROUND: The current NIH and French consensus provide physicians with clear guidelines on how to care best for patients with hepatitis C. METHODS: Review and discussion. RESULTS: Confirming the diagnosis and guiding the initial investigations have become straightforward. The standard treatment and its monitoring have been described in many publications. Recommending therapy to patients with moderate fibrosis has been the custom since the 1999 EASL guidelines. The 2002 guidelines have widened the spectrum of patients with chronic hepatitis C that should be considered for antiviral therapy. Patient categories not previously considered for therapy, such as alcoholics, intravenous drug users, prison inmates and social subgroups of society that lack adequate medical care, can now be offered therapy provided they are well supported in specific programmes. Liver physicians have learned throughout the years to manage side effects successfully and encourage patient adherence. This is reflected in the higher sustained viral response rates with standard interferon and ribavirin reported in the pegylated interferon registration trials compared with the interferon-ribavirin trials. Reducing the dose rather than stopping therapy is the key issue. Antidepressive agents have their place in the management of mood disorders prior to or during therapy. CONCLUSION: Every patient with chronic hepatitis C should be considered for antiviral therapy. It is probably best for a patient to be treated by a physician who has experience in managing possible side effects and in coaching a patient through his 6 or 12 months of treatment. PMID- 14743893 TI - Surgical management of acute necrotizing pancreatitis: a 13-year experience and a systematic review. AB - BACKGROUND: The course of acute pancreatitis (AP) is unpredictable and can vary from mild to lethal. Mortality varies from low (<2%) in mild cases to high (20% 70%) in the case of infected pancreatic necrosis. Surgical management has not been investigated in well-designed trials. Based on literature review and retrospective results from our institution, recent insights are summarized and recommendations concerning surgical treatment of AP are given. METHODS: Data of patients who underwent necrosectomy for AP in our hospital in the period 1988 2001 were reviewed. Surgical treatment strategy was divided into open abdomen strategy (OAS) and primary closure with continuous postoperative lavage (CPL). An extensive database literature search was performed to obtain articles on surgical management of AP. Level 5 evidence articles were excluded. RESULTS: In our institution, 38 patients were treated with OAS and 21 with CPL. Mortality was high (47% in the OAS group and 33% in the CPL group). The primary cause of mortality was multiorgan failure. Only 50 manuscripts from the literature search contained useful data. Mortality of patients with OAS and CPL treatment was 27% and 15%. respectively. Fewer cases of gastro-intestinal fistulas. bleeding and re interventions were reported with CPL. The majority of all survivors regained a good quality of life. CONCLUSION: Mortality of acute necrotizing pancreatitis remains high, despite optimal surgical and medical treatment. Current surgical practice is not based on well-designed clinical trails. Randomized studies are needed to define evidence-based surgery in acute necrotizing pancreatitis. PMID- 14743894 TI - Acute idiopathic pancreatitis: does it really exist or is it a myth? AB - BACKGROUND: Acute pancreatitis is a severe disease with considerable morbidity and mortality. Gallstones and alcohol abuse are the most frequent causes (75% of patients). Other well-known causes are: hyperlipidemia, hypercalcaemia, abdominal surgery and drugs. In 10%-40% of patients however, no cause is identified after initial diagnostic evaluation: acute idiopathic pancreatitis. Identifying a cause in these patients is important, since the recurrence rate is high. METHODS: A systematic review of the current literature was performed to identify possible causes, diagnoses and treatment options of acute idiopathic pancreatitis. Relevant literature was found via Pubmed. RESULTS: The presence of microlithiasis or biliary sludge is an important cause of acute 'idiopathic' pancreatitis (up to 80% of patients). Microlithiasis and sludge can be detected by transabdominal/endoscopic ultrasonography, ERCP or polarizing light microscopy of bile. Cholecystectomy is the treatment of choice, whereas endoscopic sphincterotomy and ursodeoxycholic acid maintenance therapy are effective alternatives. Sphincter of Oddi dysfunction can be identified as the cause of acute 'idiopathic' pancreatitis in up to 30% of patients. Manometry of Oddi's sphincter is the gold standard for its diagnosis. Endoscopic sphincterotomy prevents recurrence in most patients. Anatomic abnormalities such as major papilla stenosis, pancreas divisum, pancreatic duct strictures and tumours may also cause acute 'idiopathic' pancreatitis. Endoscopic sphincterotomy and surgery are effective treatments. Finally, genetic screening may reveal gene mutations as the cause of acute 'idiopathic' pancreatitis. CONCLUSIONS: Acute 'idiopathic' pancreatitis is a severe disease with a high recurrence rate. Extensive diagnostic investigations may lead to a cause in >90% of patients. PMID- 14743895 TI - Variations of the superior cerebellar artery: MR angiographic demonstration. AB - PURPOSE: The purpose of this study was to describe variations of the proximal segments of the superior cerebellar artery (SCA) detected by magnetic resonance (MR) angiography. MATERIALS AND METHODS: We reviewed 145 consecutive MR angiograms. All patients were studied with a 1.5-Tesla imager using the three dimensional time-of-flight technique. RESULTS: There were 16 duplicated SCAs in 13 patients, seven SCAs originating from the posterior cerebral arteries in six patients, four early bifurcations of the SCAs in four patients, and one SCA arising from the internal carotid artery. Because the SCA is small in caliber, the bilateral SCAs in nine patients could not be identified on MR angiograms owing to patient movement. In two patients with duplicated SCA, one of the duplicated trunks compressed the trigeminal nerve at the root entry zone, resulting in trigeminal neuralgia. CONCLUSION: Although most of these SCA variations have no clinical significance, preoperative identification of SCA variations is important for avoiding complications during surgery and/or for interventional procedures of the distal basilar artery. PMID- 14743896 TI - Evaluation of hypervascular hepatocellular carcinoma in cirrhotic liver by means of helical CT: comparison of different contrast medium concentrations within the same patient. AB - PURPOSE: To compare enhancement of the aorta, portal vein, liver, and tumor with contrast medium of a higher iodine concentration to that with one of a standard iodine concentration for liver dynamic CT within the same patient with known liver cirrhosis and hepatocellular carcinoma (HCC). MATERIALS AND METHODS: Fifty three patients with known cirrhosis and HCC underwent repeat computed tomographic (CT) examinations within three months, the first with either 300 (n=26) or 370 mgI/ml (n=27) of nonionic contrast material. In the second examination, only concentration was altered. In 28 patients, weight was equal to or more than 65 kg, and in 25 patients it was less than 65 kg. The CT numbers (HU) of the aorta, portal vein, liver, and tumor were obtained, and CT attenuation was compared between the two concentration studies. The degree of enhancement was scored qualitatively. RESULTS: Mean enhancement values of the aorta, liver, portal vein, and tumor were greater with the 370 mgI/ml injections than with the 300 mgI/ml injections throughout the study. In visual analysis, the difference in aortic, portal venous, liver, and tumor enhancement was not statistically significant between the two groups in patients weighing less than 65 kg. However, in patients weighing 65 kg or more, strong aortic and portal venous enhancement (rated as good or excellent) occurred more frequently with the 370 mgI/ml injections than with the 300 mgI/ml injections. CONCLUSION: Higher contrast enhancement was achieved in patients who received 370 mgI/ml of contrast material, resulting in better tissue contrast between liver parenchyma and HCCs. However, this difference was not visually significant in patients weighing less than 65 kg. PMID- 14743897 TI - CT perfusion of the liver during selective hepatic arteriography: pure arterial blood perfusion of liver tumor and parenchyma. AB - PURPOSE: To quantify pure arterial blood perfusion of liver tumor and parenchyma by using CT perfusion during selective hepatic arteriography. METHODS: A total of 44 patients underwent liver CT perfusion study by injection of contrast medium via the hepatic artery. CT-perfusion parameters including arterial blood flow, arterial blood volume, and arterial mean transit time in the liver parenchyma and liver tumor were calculated using the deconvolution method. The CT-perfusion parameters and vascularity of the tumor were compared. RESULTS: A complete analysis could be performed in 36 of the 44 patients. For liver tumor and liver parenchyma, respectively, arterial blood flow was 184.6 +/- 132.7 and 41.0 +/- 27.0 ml/min/ 100 g, arterial blood volume was 19.4 +/- 14.6 and 4.8 +/- 4.2 ml/100 g, and arterial mean transit time was 8.9 +/- 4.2 and 10.2 +/- 5.3 sec. Arterial blood flow and arterial blood volume correlated significantly with the vascularity of the tumor; however no correlation was detected between arterial mean transit time and the vascularity of the tumor. CONCLUSION: This technique could be used to quantify pure hepatic arterial blood perfusion. PMID- 14743898 TI - Communicating vein between the left renal vein and left ascending lumber vein: incidence and significance on abdominal CT. AB - PURPOSE: To examine the incidence and appearance of the communicating vein between the left renal vein (LRV) and the left ascending lumbar vein (LALV) on abdominal CT. MATERIALS AND METHODS: We reviewed the appearance of the communicating vein on contrast-enhanced CT obtained by multidetector-row CT (MDCT) scanner. One hundred patients without known abnormality in this region were randomly chosen and the following findings were recorded: (1) presence or absence of the visible communicating vein, (2) diameter of the communicating vein, (3) visible length of the vein (none, partial, or total), (4) laterality of the ascending lumbar vein, (5) distance between the superior mesenteric artery and the aorta. RESULTS: The communicating vein was visible in 35 patients (35%). In 20 cases, this vein was visualized within the paraaortic region and could not be traced toward the LALV. The distance between the superior mesenteric artery and the aorta was narrower in the patients with visible communicating vein than in those without it, however, no statistically significant difference was demonstrated. CONCLUSION: This vein is commonly visible in the general population, and care should be taken not to confuse it with lymphadenopathy because this communicating vein was partially visualized within the paraaortic region in 20% of the cases. PMID- 14743899 TI - Crackle analysis for chest auscultation and comparison with high-resolution CT findings. AB - PURPOSE: The purpose of our study was to clarify the correlation between respiratory sounds and the high-resolution CT (HRCT) findings of lung diseases. MATERIALS AND METHODS: Respiratory sounds were recorded using a stethoscope in 41 patients with crackles. All had undergone inspiratory and expiratory CT. Subjects included 18 patients with interstitial pneumonia and 23 without interstitial pneumonia. Two parameters, two-cycle duration (2CD) and initial deflection width (IDW) of the "crackle," were induced by time-expanded waveform analysis. Two radiologists independently assessed 11 HRCT findings. An evaluation was carried out to determine whether there was a significant difference in the two parameters between the presence and absence of each HRCT finding. RESULTS: The two parameters of crackles were significantly shorter in the interstitial pneumonia group than the non-interstitial pneumonia group. Ground-glass opacity, honeycombing, lung volume reduction, traction bronchiectasis, centrilobular nodules, emphysematous change, and attenuation and volume change between inspiratory and expiratory CT were correlated with one or two parameters in all patients, whereas the other three findings were not. Among the interstitial pneumonia group, traction bronchiectasis, emphysematous change, and attenuation and volume change between inspiratory and expiratory CT were significantly correlated with one or two parameters. CONCLUSION: Abnormal respiratory sounds were correlated with some HRCT findings. PMID- 14743900 TI - Coronal multiplanar reconstruction view from whole lung thin-section CT by multidetector-row CT: determination of malignant or benign lesions and differential diagnosis in 68 cases of solitary pulmonary nodule. AB - PURPOSE: The purpose of this study was to evaluate the usefulness of the coronal multiplanar reconstruction (MPR) view in comparison with transverse helical thin section CT for both the determination of malignant or benign lesions and the differential diagnosis of solitary pulmonary nodules. MATERIALS AND METHODS: Sixty-eight cases of pathologically proved solitary pulmonary nodule less than 3 cm in diameter were enrolled in this study. For the routine study, transverse helical thin-section CT (1.25 mm collimation, FOV 20 cm) covering the areas with solitary pulmonary nodules as well as whole lung helical thin-section CT (2.5 mm collimation, 1.25 mm reconstruction interval, FOV 34.5 cm, pitch 6:1, high spatial frequency algorithm) were scanned with a multidetector-row CT (MDCT) scanner. From the whole lung thin-section CT data, coronal MPR views (2.5 mm slice thickness) were reconstructed on a workstation. ROC analysis was used for an observer performance study, in which three observers indicated their confidence level for the determination of malignant or benign lesion for the nodules by means of transverse thin-section CT and coronal MPR. In addition, the observers recorded appropriate disease entities as the final diagnosis of each case. Accuracies of the final diagnosis based on the two sets of images were compared with McNemer' s test. RESULTS: In terms of the determination of malignant or benign lesion, there was no significant difference between the two sets of images (coronal MPR and transverse thin-section CT; mean Az=0.853 and 0.854, respectively). In addition, accuracy of the final diagnosis based on coronal MPR views (74%) was almost equal to that based on transverse thin-section CT (71%) (p=0.3). CONCLUSIONS: The diagnostic efficacy of the coronal MPR view is comparable to that of transverse thin-section CT for the evaluation of solitary pulmonary nodules. PMID- 14743901 TI - False-positive CT and thallium-201 SPECT results in a patient with suspected lung cancer and mediastinal lymph node metastasis. AB - We describe a patient with a final diagnosis of inflammatory change in which combined imaging results were strongly suggestive of lung cancer with mediastinal lymph node metastases. The patient, who was suspected of having a pulmonary lesion on chest radiography performed at a local hospital, was referred to our hospital. Contrast-enhanced computed tomography (CT) of the chest showed a small sized opacity in the left upper lung and multiple mediastinal nodules. The largest mediastinal nodule, depicted at a pretracheal region, was 22 mm in size. Thallium-201 single photon emission computed tomography (SPECT) showed increased uptake in these lesions, which exhibited no remarkable washout of the tracer, supportive of the diagnosis of lung cancer with mediastinal lymph node metastasis. However, bronchoscopic and mediastinoscopic biopsy revealed no malignant cells or reactive lymph nodes, respectively. Therefore, no surgical procedure was performed after the biopsy. The patient has done well without evidence of malignancy for 21 months. Radiologists should be careful in the differential diagnosis of benign and malignant lesions even with combined imaging modalities. PMID- 14743902 TI - Persistent Mullerian duct syndrome suggested by magnetic resonance imaging. AB - We report the case of a 45-year-old man with persistent Mullerian duct syndrome suggested by its characteristic appearance magnetic resonance (MR) imaging. A tubular structure extending from the patient's left inguinal region to the pelvic cavity was found incidentally at surgery for a testicular tumor. T2-weighted MR images obtained after surgery revealed the characteristic three-layer structure in the left pelvic cavity with no relation to the bladder or prostate. The hyperintense inner layer, hypointense middle layer, and hyperintense outer layer were thought to represent the endometrium, junctional zone, and outer myometrium of the uterus, respectively. Cryptorchidism and transverse testicular ectopia were not associated. The structure was not resected because there had been no reports of malignancy arising from the Mullerian duct remnant. PMID- 14743903 TI - Real-time estimation system for mean glandular dose in mammography. AB - PURPOSE: The purpose of this study was to construct a system that calculates mean glandular dose (MGD) in mammography in real-time on the RIS network. MATERIALS AND METHODS: Measurement of the half-value layer (HVL) and exposure dose were measured according to the Japanese mammographic quality control manual. A parallel plate chamber (10X5-6M; soft X-rays only) was used for measurement. Exposure dose was measured from 26 to 32 kV. RESULTS: Effective energy (Eeff [keV]) could be determined from tube voltage (V [kV] ) using the measure of the HVL according to the following equations: for the Mo/Mo combination, Eeff = 0.1325V + 11.80; for the Mo/Rh combination, Eeff = 0.1435V + 12.20; for the Rh/Rh combination, Eeff = 0.1724V + 11.29. Exposure output (XmAs [mR/mAs]), which can be determined from dosimetry per 1 mAs, becomes the following equation if effective energy is expressed as a variable: for the Mo/Mo combination, XmAs = 2.1329 Eeff2 - 57.784 Eeff + 392.71; for the Mo/Rh combination, XmAs = 1.1919 Eeff2 - 31.924 Eeff + 212.23; and for the Rh/Rh combination, XmAs = 2.6929 Eeff2 82.831 Eeff + 643.54. The MGD conversion factor (DgN [mGy/mR]) can be determined from effective energy using the ACR protocol according to the following equation: for the Mo/Mo combination, DgN = (0.3962 Eeff- 4.3178) 10(-3); for the Mo/Rh combination, DgN = (0.3495 Eeff - 3.5479) x 10(-3); and for the Rh/Rh combination, DgN = (0.4498 Eeff - 5.0448) x 10(-3). However, in these cases, compression breast thickness (CBT) was 4.2 cm, and the ratio of adipose and glandular tissue was 50/50. CONCLUSION: MGD can be determined using mAs obtained from mammography, exposure output (XmAs), MGD conversion factor (DgN), and source skin distance compensation factor (SSDcf) according to the following equation: MGD [mGy] = mAs x XmAs [mR/mAs] x DgN [mGy/mR] x SSDcf where SSDcf = (64/(64 - CBT [cm]))2. PMID- 14743904 TI - Regulation of neurosurgical innovation. PMID- 14743905 TI - Surgical innovation or surgical evolution: an ethical and practical guide to handling novel neurosurgical procedures. AB - OBJECT: Surgical innovation is an important driver of improvements in technique and technology, which ultimately translates into improvements in patients' outcomes. Nevertheless, patients may face new risks of morbidity and mortality when surgical innovation is used, and well-intentioned surgical "experimentation" on patients must be regulated and monitored. In this paper the authors examine the challenges of defining surgical innovation and briefly review the literature on this challenging subject. METHODS: Using examples from the field of neurosurgery and in part from the personal experience of the senior author, the authors develop a model of levels of experimental acuity of surgical procedures and offer recommendations on how these procedures would best be regulated. CONCLUSIONS: The authors propose guidelines for determining the need for regulation of innovation. The potential role of institutional review boards in this process is highlighted. PMID- 14743906 TI - Delayed neurological deficits detected by an ischemic pattern in the extracellular cerebral metabolites in patients with aneurysmal subarachnoid hemorrhage. AB - OBJECT: In the treatment of patients with aneurysmal subarachnoid hemorrhage (SAH), early occlusion of the aneurysm is necessary as well as monitoring and treatment of complications following the primary bleeding episode. Monitoring with microdialysis has been studied for its ability to indicate and predict the occurrence of delayed ischemic neurological deficits (DINDs) in patients with SAH. METHODS: In 42 patients with aneurysmal SAH microdialysis monitoring of metabolites was performed using a 0.3-microl/minute perfusion flow over several days, and the results were correlated to clinical events and to brain infarction observed on computerized tomography scans. The microdialysis probe was inserted into the territory of the parent artery of the aneurysm. The authors defined an ischemic pattern as increases in the lactate/glucose (L/G) and lactate/pyruvate (L/P) ratios that were greater than 20% followed by a 20% increase in glycerol concentration. This ischemic pattern was found in 17 of 18 patients who experienced a DIND and in three of 24 patients who did not experience a delayed clinical deterioration. The ischemic pattern preceded the occurrence of a DIND by a mean interval of 11 hours. Maximum L/G and L/P ratios did not correlate with the presence of DIND or outcome, and there was no association between the glycerol level and subsequent brain infarction. CONCLUSIONS: Microdialysis monitoring of the cerebral metabolism in patients with SAH may predict with high sensitivity and specificity the occurrence of a DIND. Whether an earlier diagnosis results in better treatment of DINDs and, therefore, in overall better outcomes remains to be proven, as it is linked to an efficacious treatment of cerebral vasospasm. PMID- 14743907 TI - The multichannel auditory brainstem implant: how many electrodes make sense? AB - OBJECT: Development of multichannel auditory brainstem implant (ABI) systems has been based in part on the assumption that audiological outcome can be optimized by increasing the number of available electrodes. In this paper the authors critically analyze this assumption on the basis of a retrospective clinical study performed using the Nucleus 22 ABI surface electrode array. METHODS: The perceptual performances of 61 patients with neurofibromatosis Type 2 were tested approximately 6 weeks after an eight-electrode ABI had been implanted. Of eight implanted electrodes 5.57 +/- 2.57 (mean +/- standard deviation [SD] provided auditory sensations when stimulated. Electrodes were deactivated when stimulation resulted in significant nonauditory side effects or no auditory sensation at all, and also when they failed to provide distinctive pitch sensations. The mean (+/- SD) scores for patients with ABIs were the following: sound-only consonant recognition, 20.4 +/- 14.3 (range 0-65%); vowel recognition, 28.8 +/- 18% (range 0-67%); Monosyllable Trochee Spondee (MTS) word recognition 41.1 +/- 25.3% (range 0-100%); and sentence recognition, 5.3 +/- 11.4% (range 0-64%). Performance in patients in whom between one and three electrodes provided auditory sensation was significantly poorer than that in patients with between four and eight functional electrodes in the vowel, MTS word, and City University of New York (CUNY) sentence recognition tests. The correlation between performance and electrode number did not reach the 0.05 level of significance with respect to the sound effect, consonant, and MTS stress-pattern recognition tests, probably because a satisfactory performance in these tests can be obtained only with temporal cues, that is, without any information about the frequency of the sounds. In the MTS word and the CUNY sentence recognition tests, performance was optimal in the patients with eight functional electrodes. Although all top performers had more than three functional auditory electrodes, no further improvement (asymptotic performance) was seen in those with five or more active electrodes in the consonant, vowel, and sound effect recognition tests. CONCLUSIONS: A minimum of three spectral channels, programmed in the appropriate individual tonotopic order seem to be required for satisfactory speech recognition in most patients with ABI. Due to the limited access to the tonotopic frequency gradient of the cochlear nucleus with surface stimulation, patients with ABI do not receive a wide range of spectral cues (frequency information) with multielectrode (> 5) surface arrays. PMID- 14743908 TI - Inflammation markers and risk factors for recurrence in 35 patients with a posttraumatic chronic subdural hematoma: a prospective study. AB - OBJECT: To evaluate the role of local inflammation in the pathogenesis and postoperative recurrence of chronic subdural hematoma (CSDH), the authors conducted an investigation in a selected group of patients who could clearly recall a traumatic event and who did not have other risk factors for CSDH. Inflammation was analyzed by measuring the concentration of the proinflammatory and inflammatory cytokines interleukin (IL)-6 and IL-8. The authors also investigated the possible relationship between high levels of local inflammation that were measured and recurrence of the CSDH. METHODS: A prospective study was performed between 1999 and 2001. Thirty-five patients who could clearly recall a traumatic event that had occurred at least 3 weeks previously and who did not have risk factors for CSDH were enrolled. All patients were surgically treated by burr hole irrigation plus external drainage. The concentration of inflammatory cytokines was very high in the lesion, whereas it was normal in serum. In five cases in which recurrence occurred, concentrations of both IL-6 and IL-8 were significantly increased (p < 0.01) in comparison with cases without a recurrence. In a layering hematoma, the IL-6 and IL-8 concentrations were significantly higher (p < 0.05). Layering CSDHs were also significantly correlated with recurrence. Trabecular hematoma had the lowest cytokine levels and the longest median interval between trauma and clinical onset. The interval from trauma did not significantly influence recurrence, although it did differ significantly between the trabecular and layering CSDH groups. Concentrations of IL-6 and IL-8 in the CSDHs did not differ significantly in relation to either the age of the hematoma (measured as the interval from trauma) or the age of the patient. CONCLUSIONS: Brain trauma causes the onset of an inflammatory process within the dural border cell layer; high levels of inflammatory cytokines were significantly correlated with recurrence and layering CSDH. A prolonged postoperative antiinflammatory medicine given as prophylaxis may help prevent the recurrence of a CSDH. PMID- 14743909 TI - Surgical treatment of symptomatic Rathke cleft cysts: clinical features and results with special attention to recurrence. AB - OBJECT: Rathke cleft cysts (RCCs) are rarely symptomatic. The purpose of this study was to clarify the clinical, neuroimaging, surgical, and pathological features of symptomatic RCCs with special attention to their recurrence. METHODS: This retrospective study involved 53 patients with pathologically confirmed symptomatic RCCs. There were 28 female and 25 male patients, ranging in age from 11 to 68 years (mean 37 years). Common clinical presentations included headache, visual impairment, and endocrine disturbance. The most common endocrine disturbances were hyperprolactinemia and diabetes insipidus. Most of these improved or were resolved after surgery, with the exception of diabetes insipidus and panhypopituitarism. The cysts were intrasellar with suprasellar extension in 33 patients, and ranged in size from 5 to 40 mm (mean 17 mm). In the 50 magnetic resonance (MR) images that were reviewed, the signal intensities were quite variable. Fourteen MR images demonstrated enhancement after an injection of Gd. Intraoperatively, the cyst contents were found to be yellowish (18 patients [37%]) and mucoid (25 patients [51%]). Pathological examinations revealed a pseudostratified columnar epithelium in 26 patients (49%). Abundant squamous metaplasia and a stratified squamous epithelium were also found in 12 patients (23%). Follow-up MR images revealed cyst recurrences that required a repeated operation in six patients. Statistically significant risk factors for a recurrence included enhancement of the lesion on MR images (p = 0.017), the extent of cyst removal (p = 0.012), and the presence of squamous epithelium (p = 0.008). CONCLUSIONS: Rathke cleft cysts are associated with a variety of clinical presentations and sometimes confusing intraoperative and pathological findings. Close postoperative observation with neuroimaging and neuroophthalmological assessment is necessary, especially after a partial removal, as in cases with squamous metaplasia. PMID- 14743910 TI - Volume of residual disease as a predictor of outcome in adult patients with recurrent supratentorial glioblastomas multiforme who are undergoing chemotherapy. AB - OBJECT: For patients with recurrent glioblastomas multiforme (GBMs) the prognosis is poor. Although chemotherapy may provide a survival advantage, the role of the extent of tumor resection, or the volume of the residual tumor at the time of recurrence, before instituting chemotherapy, is unclear. This study was designed to assess the response to chemotherapy based on the volume of residual disease (VRD) at the start of treatment in patients with recurrent GBMs. To accomplish this, the authors evaluated a homogeneous group of patients with recurrent GBMs who received the same chemotherapeutic agent. METHODS: One hundred nineteen adult patients with recurrent supratentorial GBMs received temozolomide chemotherapy at the time of tumor recurrence. In this cohort the authors analyzed the prognostic significance of volumetrically assessed tumor mass on time to tumor progression (TTP) and survival time (ST). Multivariate analysis demonstrated that the VRD at the beginning of chemotherapy was a statistically significant predictor of both TTP (p < 0.0001) and ST (p < 0.006) when adjusted for the patient's age, performance score, and time from the initial diagnosis. Patients in whom the VRD was less than 10 cm3 at the start of chemotherapy had a 6-month progression-free survival rate of 32% compared with 8% for patients with a VRD between 10 and 15 cm3 and 3% for patients with a VRD larger than 15 cm3. Patients in whom the VRD was smaller than 10 cm3 had a 1-year survival rate of 37% compared with 9% for patients with a VRD between 10 and 15 cm3 and 18% for patients with a VRD larger than 15 cm3. CONCLUSIONS: These data indicate that patients with recurrent GBMs who start chemotherapy with a smaller volume (< 10 cm3) of residual disease may have a more favorable response to chemotherapy and a more favorable outcome. PMID- 14743911 TI - Errors in compliance with federal rules and regulations relating to healthcare benefits programs: the University of Washington Department of Neurological Surgery experience. AB - This article details the errors in compliance with federal rules and regulations relating to the healthcare benefits programs at the University of Washington Department of Neurological Surgery from 1996 through 2002. University faculty members, regardless of the organization to which they belong, will be identified by the federal government as the individual responsible in healthcare finance inquiries. A full understanding of all regulations and an active compliance program are necessary to avoid problems, including criminal prosecution. PMID- 14743912 TI - Serial evaluation of axonal function in patients with brain death by using anisotropic diffusion-weighted magnetic resonance imaging. AB - OBJECT: The purposes of this study were to evaluate the serial changes in diffusion anisotropy of the brain, probably reflecting axonal function in brain dead patients, and thus to explore the possibility of quantitatively estimating the risk of brain death. METHODS: Ten patients suffering from stroke with or without impending brain death and 10 healthy volunteers were studied using three dimensional anisotropy contrast (3DAC) magnetic resonance (MR) axonography with the aid of a 1.5-tesla MR imaging system. To detect changes in the diffusion anisotropy of neural bundles, the corticospinal tract was evaluated. Diffusion anisotropy of short axonal fibers decreased immediately after apparent brain death. Whereas the trichromatic coefficients of the corticospinal tract greatly diminished between 6 and 12 hours after apparent brain death, the coefficients of the corpus callosum and the optic radiation decreased in less time, that is, between 1 and 6 hours. The coefficients of these three bundles turned isotropic between 24 and 44 hours after apparent brain death. CONCLUSIONS: Results of 3DAC MR axonography revealed that diffusion anisotropy of neural bundles diminished between 1 and 12 hours after the onset of apparent brain death, probably depending on the length of the bundles, and disappeared between 24 and 44 hours after the onset of brain death, which might reflect dynamic changes of axonal structure and indirectly herald axonal dysfunction. These findings seem to be greatly helpful in establishing an appropriate method to estimate the risk of brain death quantitatively and in forming the basis of future definitions of brain death. PMID- 14743914 TI - Transcranial echo-guided transsphenoidal surgical approach for the removal of large macroadenomas. AB - OBJECT: Transsphenoidal surgery for the removal of macroadenomas has some disadvantages, including the risk of performing procedures without adequate visualization, difficulties in estimating the amount of residual tumor, and the risk of injuring major vessels. To overcome these disadvantages, the authors have developed transcranial echo-guided transsphenoidal surgery. METHODS: Three patients with large macroadenomas and two patients with irregularly shaped macroadenomas were selected for this operation. In addition to standard preparations for transsphenoidal surgery, in each case the right frontal bone was trephined and an echo probe was inserted transdurally through the trephination hole. During tumor removal, brightness-mode echo images and Doppler color flow images were obtained. The echo images allowed for real-time visualization of the tumor and surrounding brain structures including major arteries and the cisterns; histological heterogeneities of the tumor could also be appreciated. The tumors were removed safely and maximal tumor removal was achieved. CONCLUSIONS: Transcranial echo-guided transsphenoidal surgery provides real-time visualization of tumor removal. The method enhances the safety of this surgery, maximizes the removal of the tumor, and is inexpensive. PMID- 14743913 TI - Cystic glioblastoma multiforme: survival outcomes in 22 cases. AB - OBJECT: The goal of this study was to determine whether the presence of a large tumor cyst was associated with improved outcome in patients undergoing surgery for newly diagnosed glioblastomas multiforme (GBMs) by comparing these patients with a matched cohort of patients with noncystic GBMs in clinical features, tumor imaging characteristics, survival, and time to tumor recurrence after surgery. METHODS: A retrospective analysis was conducted in 22 patients by using imaging information and chart reviews of operative reports of GBMs with large cysts (> or = 50% of tumor volume) at The University of Texas M. D. Anderson Cancer Center between 1993 and 2002. Clinical and neurosurgical outcomes and recurrence rates were studied. A statistical comparison was made with a matching cohort of 22 patients with noncystic GBMs. No significant differences in clinical variables were found between the cohort with cystic GBMs and the matched cohort with noncystic GBMs. To avoid bias in preoperative assessment of tumor volume, the tumor burden was compared in patients whose tumors had cysts (excluding the cystic mass) and in patients whose tumors did not contain cysts. There was no statistically significant difference between the two groups (p = 0.8). In patients with cystic GBMs the median survival time after surgery was 18.2 months (95% confidence interval [CI] 11.9-24.5 months) and at 2 years 43% of the patients were still alive. In comparison, in patients with noncystic GBMs, the median survival time was 14.3 months (95% CI 12.1-16.4 months) and only 16% of patients were alive at 2 years. The median time to tumor recurrence was 7.6 months (95% CI 0.01-18 months) in patients harboring cystic GBMs and 4.2 months (95% CI 1.8-6.6 months) in the matched cohort (log-rank test, p = 0.04). In the cystic GBM group, no recurrence was observed in 53% of patients at 6 months, 45% at 1 year, and 38% at 2 years after surgery, whereas the corresponding numbers for the noncystic group were 36, 14, and 9%, respectively. CONCLUSIONS: The results indicate that patients harboring a GBM that contains a large cyst survive longer and have a longer time to recurrence than those who lack such a cyst. This is the first such observation in the literature. PMID- 14743915 TI - What we research: survey of American Association of Neurological Surgeons and Congress of Neurological Surgeons member publications. AB - OBJECT: The goal of this study was to create a searchable database of research manuscripts authored by members of the American Association of Neurological Surgeons and the Congress of Neurological Surgeons (AANS/CNS) to describe the nature and character of the research currently being undertaken by neurosurgeons. METHODS: Manuscripts published by all physician members listed in the 2001 AANS/CNS Membership Directory (6921 physicians) were gathered into a database through individual literature searches of the author name for the calendar year 2001. Duplicate publications were purged and the database was reviewed for accuracy. An internal verification of the database revealed a 4% underreporting rate. Statistics from the database were compiled and displayed with information about AANS/CNS members and their clinical activities. The AANS/CNS members published a total of 2748 research the manuscripts in 479 different journals during 2001. Thirty-eight percent of the manuscripts (1042 of 2748) were authored by US members and 62% (1706 of 2748) by non-US members. The focus of the majority of manuscripts included the areas of brain tumor (26%; 707 of 2748), vascular disease (20%; 558 of 2748), spine (10%; 282 of 2748), and trauma (8%; 233 of 2748). Sixty-nine percent of manuscripts (1897 of 2748) were retrospective and technical clinical studies, and of these 39% (744 of 1897) were case reports. Laboratory investigations made up 15% (414 of 2748) of all manuscripts, whereas prospective randomized clinical trials represented 1% (34 of 2748). CONCLUSIONS: The majority of AANS/CNS member manuscripts are authored by non-US members despite their small AANS/CNS representation. Most research is clinical, based on retrospective data, and includes a large number of case reports. A disparity exists between what neurosurgeons do clinically and both the quantity and subject of their research. PMID- 14743916 TI - Prevention of apoptotic but not necrotic cell death following neuronal injury by neurotrophins signaling through the tyrosine kinase receptor. AB - OBJECT: Neurotrophins prevent the death of neurons during embryonal development and have potential as therapeutic agents. During development, neuronal death occurs only by apoptosis and not by necrosis. Following injury, however, neurons can die by both processes. Data from prior studies have not clearly indicated whether neurotrophins can decrease apoptosis compared with necrosis. The goal of this study was to determine the effect of neurotrophin treatment on each of these processes following injury and to characterize the receptor(s) required. METHODS: The authors used an in vitro model of injury with the aid of primary cortical neurons obtained from rat embryos. After 9 days in culture and the elimination of glia, homogeneous and mature neurons were available for experimentation. Noxious stimuli were applied, including radiation, hypoxia, and ischemia. Subsequent cell death by apoptosis or necrosis was noted based on morphological and enzymatic assessments (such as lactate dehydrogenase [LDH] release) and assays for DNA fragmentation. The effect of treatment with nerve growth factor (NGF), brain derived neurotrophic factor (BDNF), and neurotrophin-3 was determined. Finally, Western blot analyses were performed to note the neurotrophin receptor status in the neurons (tyrosine kinase receptors [Trks] and p75). The authors studied different stimuli-induced cell death by using different processes. With the application of radiation, cells died primarily by apoptosis, as evidenced by cell shrinkage, the presence of apoptotic bodies, and specific DNA fragmentation. This was a delayed process (> 6 hours) that could be reduced by gene transcription or protein synthesis inhibitors. With ischemia, cells died immediately by necrosis, showing cell enlargement and rupture. Ischemic cell death was not affected by the inhibition of macromolecular synthesis. Hypoxia produced a mixture of the two cell death processes. Both BDNF and neurotrophin-3 demonstrated protection against apoptotic cell death only. Statistically significant decreases of both LDH release and apoptosis-specific DNA fragmentation were noted following radiation and hypoxia, but not for ischemia. Nerve growth factor, unlike the other neurotrophins, did not affect apoptosis because a functional receptor, Trk A, was not expressed by the cortical neurons. There was expression of both Trk B and Trk C, which bind BDNF and neurotrophin-3. CONCLUSIONS: These findings have significant clinical implications. Neurotrophins may only be effective in disorders in which apoptosis, and not necrosis, is the major process. Furthermore, the Trk signaling cascade must be activated for this response to occur. Because the expression of these receptors diminishes in adulthood, neurotrophin application may be most appropriate in the pediatric population. PMID- 14743917 TI - Proliferation, migration, and differentiation of human neural stem/progenitor cells after transplantation into a rat model of traumatic brain injury. AB - OBJECT: Cultures containing human neural stem and progenitor cells (neurospheres) have the capacity to proliferate and differentiate into the major phenotypes of the adult brain. These properties make them candidates for therapeutic transplantation in cases of neurological diseases that involve cell loss. In this study, long-term cultured and cryopreserved cells were transplanted into the traumatically injured rat brain to evaluate the potential for human neural stem/progenitor cells to survive and differentiate following traumatic injury. METHODS: Neural stem/progenitor cell cultures were established from 10-week-old human forebrain. Immunosuppressed adult rats received a unilateral parietal cortical contusion injury, which was delivered using the weight-drop method. Immediately following the injury, these animals received transplants of neural stem/progenitor cells, which were placed close to the site of injury. Two or 6 weeks after the procedure, these animals were killed and their brains were examined by immunohistochemical analysis. At both 2 and 6 weeks postoperatively, the transplanted human cells were found in the perilesional zone, hippocampus, corpus callosum, and ipsilateral subependymal zone of the rats. Compared with the 2-week time point, an increased number of HuN-positive cells was observed at 6 weeks. In addition, at 6 weeks post-injury/transplantation, the cells were noted to cross the midline to the contralateral corpus callosum and into the contralateral cortex. Double labeling demonstrated neuronal and astrocytic, but not oligodendrocytic differentiation. Moreover, the cortex appeared to provide an environment that was less hospitable to neuronal differentiation than the hippocampus. CONCLUSIONS: This study shows that expandable human neural stem/progenitor cells survive transplantation, and migrate, differentiate, and proliferate in the injured brain. These cells could potentially be developed for transplantation therapy in cases of traumatic brain injury. PMID- 14743918 TI - Proximal occlusion of the middle cerebral artery in C57Black6 mice: relationship of patency of the posterior communicating artery, infarct evolution, and animal survival. AB - OBJECT: The intraluminal suture model for focal cerebral ischemia is increasingly used, but not without problems. It causes hypothalamic injury, subarachnoid hemorrhage, and inadvertent premature reperfusion. The patency of the posterior communicating artery (PCoA) potentially affects the size of the infarct. In addition, survival at 1 week is unstable. The authors operated on C57Black6 mice to produce proximal middle cerebral artery occlusion (MCAO) so that drawbacks with the suture model could be circumvented. METHODS: The MCA segment just proximal to the olfactory branch was occluded either permanently or temporarily. After 1 hour of MCAO the infarct volume was significantly smaller than that found after 2 hours or in instances of permanent MCAO. The differences were assessed at 24 hours and 7 days after surgery (p < 0.05 and p < 0.001, respectively). The patency of the PCoA, as visualized using carbon black solution, did not correlate with the infarct size. Neurologically, the 1- and 2-hour MCAO groups displayed significantly less severe deficits than the permanent MCAO group on Days 1, 4, and 7 (p < 0.005 and p < 0.01, respectively). Although the infarct size, neurological deficits, and body weight loss were more severe in the permanent MCAO group, the survival rate at Day 7 was 80%. CONCLUSIONS: This model provides not only a robust infarct size (which is not affected by the patency of the PCoA), but also a better survival rate. PMID- 14743920 TI - Tensile strength of cranial pia mater: preliminary results. AB - OBJECT: The goal of this study was to determine the tensile strength of cranial pia mater. METHODS: Samples of isolated bovine cranial pia mater were subjected to quasistatic traction to evaluate its tensile strength. The experimental curves of physiological deformation that were obtained can be subdivided into three parts that represent different mechanical properties: the nonlinear initial part of the curve demonstrates increasing stiffness, followed by a quasilinear pattern of elastic behavior, and finally a negative relationship (slope) between force and elongation, which characterizes a progressive deterioration. These three steps precede final sample rupture. The stiffness of the pia mater was calculated for both the initial and the linear (elastic) parts of the mean curve. The initial part and the elastic part of the curve show a typical stiffness value of 0.024 N/mm and 0.19 N/mm, respectively. The maximal mean force and corresponding maximal deformation that were attained were 1.1 N and 0.19, respectively. CONCLUSIONS: Although very thin and apparently fragile, pia mater exhibits an unexpectedly high level of stiffness and should have a significant influence on total brain mechanical properties in response to loading. PMID- 14743919 TI - Magnesium and experimental vasospasm. AB - OBJECT: Interest has developed in the use of magnesium (Mg++) as a neuroprotectant and antivasospastic agent. Magnesium may increase cerebral blood flow (CBF) and reduce the contraction of cerebral arteries caused by various stimuli. In this study the authors tested the hypothesis that a continuous intravenous infusion of Mg++ reduces cerebral vasospasm after experimental subarachnoid hemorrhage (SAH). METHODS: A dose-finding study was conducted in five monkeys (Macaca fascicularis) to determine what doses of intravenous MgSO4 elevate the cerebrospinal fluid (CSF) concentrations of Mg++ to vasoactive levels and to determine what effects these doses have on the diameters of cerebral arteries, as shown angiographically. After a standard dose of MgSO4 had been selected it was then administered in a randomized, controlled, blinded study to 10 monkeys (five animals/group) with SAH, beginning on Day 0 and continuing for 7 days, at which time angiography was repeated. A 0.086-g/kg bolus of MgSO4 followed by an infusion of 0.028 g/kg/day MgSO4 significantly elevated serum and CSF levels of Mg++ (five monkeys). Magnesium sulfate significantly elevated the serum level of total Mg++ from a control value of 0.83 +/- 0.04 mmol/L to 2.42 +/ 1.01 mmol/L on Day 7 and raised the CSF level from 1.3 +/- 0.04 mmol/L to 1.76 +/- 0.14 mmol/L. There was no angiographic evidence of any effect of MgSO4 on normal cerebral arteries. After SAH, the vasospasm in the middle cerebral artery was not significantly reduced (46 +/- 8% in the MgSO4-treated group compared with 35 +/- 6% in the placebo [vehicle]-treated group, p > 0.05, unpaired t-test). CONCLUSIONS: Magnesium sulfate did not significantly reduce cerebral vasospasm after SAH in the doses tested. An investigation of SAH is warranted mainly to test whether a benefit can be achieved by neuroprotection or by augmentation of CBF by dilation of small vessels and/or collateral pathways. PMID- 14743921 TI - Traumatic aneurysm of the supraclinoid internal carotid artery and an associated carotid-cavernous fistula: vascular reconstruction performed using intravascular implantation of stents and coils. Case report. AB - This report documents the treatment of a traumatic aneurysm of the supraclinoid internal carotid artery (ICA) that was associated with a carotid-cavernous fistula (CCF), which appeared following closed head trauma. This life-threatening lesion, which is very rare, required aggressive management achieved using intravascular stents and coils. A 19-year-old man presented with severe traumatic intracerebral and subarachnoid hematoma after he had suffered a severe closed head injury in a motor vehicle accident. Cerebral angiography performed 11 days after the injury demonstrated a traumatic aneurysm and severe narrowing of the right supraclinoid ICA, which was consistent with a dissection-induced stenosis associated with a direct CCF. Both lesions were successfully obliterated with preservation of the parent artery by using stents in conjunction with coils. Follow-up angiography obtained 7 months postoperatively revealed persistent obliteration of the aneurysm and CCF as well as patency of the parent artery. The patient remained asymptomatic during the clinical follow-up period of 14 months. Endovascular treatment involving the use of a stent combined with coils appears to be a feasible, minimally invasive option for treatment of this hard-to-treat lesion. PMID- 14743922 TI - Endovascular treatment of a transverse-sigmoid sinus aneurysm presenting as pulsatile tinnitus. Case report. AB - The authors report on the case of a 38-year-old woman who had experienced incapacitating pulsatile tinnitus in the left ear for 6 months. Angiographic studies revealed a wide-necked venous aneurysm of the left transverse-sigmoid sinus. Solitary stent placement across the aneurysm neck resulted in a slight modification in the lesion's characteristics. A second session, in which embolization with Guglielmi Detachable Coils was performed, resulted in a 100% occlusion of the aneurysm, with patency of the parent vessel and resolution of the tinnitus. PMID- 14743923 TI - Superior semicircular canal dehiscence syndrome. Case report. AB - The authors present the case of a man who had superior semicircular canal dehiscence syndrome in addition to chronic otitis media. This case is atypical because the patient coincidentally had middle ear and mastoid disease, which previously had been treated surgically. The prior ear surgery delayed the diagnosis of superior semicircular canal dehiscence syndrome and increased the complexity of the repair of the superior semicircular canal dehiscence. Superior semicircular canal dehiscence syndrome is a recently recognized syndrome resulting in acute or chronic vestibular symptoms. The diagnosis is made using history, vestibular examination, and computerized tomography studies. Neurosurgeons should be aware that patients with superior semicircular canal dehiscence syndrome who experience disabling chronic or acute vestibular symptoms can be treated using a joint neurosurgical-otological procedure through the middle cranial fossa. PMID- 14743924 TI - Superior semicircular canal dehiscence: a new indication for middle fossa craniotomy. Case report. AB - Superior semicircular canal dehiscence is a recently described condition resulting in pressure-induced vertigo in affected patients. The diagnosis is established with the appearance of characteristic electronystagmographic and neuroimaging findings. This condition is amenable to surgical treatment by resurfacing of the dehiscence in the defect in the middle cranial fossa floor with preservation of superior semicircular canal function. The authors report on the treatment of a 35-year-old man with superior semicircular canal dehiscence by a joint neurosurgical and otolaryngological team. PMID- 14743925 TI - Focal hyperperfusion after superficial temporal artery-middle cerebral artery anastomosis in a patient with moyamoya disease. Case report. AB - Superficial temporal artery-middle cerebral artery (STA-MCA) anastomosis is a standard surgical therapeutic option in patients with moyamoya disease. Most patients experience improvement in their clinical symptoms immediately after surgery. The authors report on the case of a 39-year-old man with moyamoya disease who suffered from temporary and frequent neurological deterioration after undergoing a left STA-MCA anastomosis. Hemodilution and hypervolemia therapies did not improve his course. Technetium-99m hexamethylpropyleneamine oxime single photon emission tomography scans demonstrated focal intense accumulation of the tracer in the frontal operculum on the side of the surgery. Although diffusion weighted magnetic resonance (MR) imaging demonstrated no abnormalities except for the postoperative change, perfusion-weighted MR images and MR digital subtraction angiography revealed focal hyperperfusion in the left frontal operculum where the cerebral perfusion reserve was severely disturbed preoperatively. This evidence strongly supports the notion that focal hyperperfusion observed after STA-MCA anastomosis could occur in the poor perfusion reserve area preoperatively and could cause temporary neurological deterioration. PMID- 14743926 TI - Performance of bipolar forceps during coagulation and its dependence on the tip material: a quantitative experimental assay. Technical note. AB - The aim of this study was to measure objectively the adherence of burned tissue to bipolar forceps to evaluate the coagulation performance of forceps made of different types of metals. Coagulation performance of bipolar forceps made of gold, titanium, and stainless steel was determined by comparing the amount of protein in the adhered coagulum on the tips. The amount of adhered coagulum was significantly less on the gold-plated bipolar forceps than on those made of the other two materials. The ease with which coagulum could be removed was compared using the cleaning cycle of an ultrasonic rinsing device. This ease of removal was also significant with the gold-plated forceps. Electron microscopy observations of the surface of the forceps tips revealed a significant difference in roughness among these materials, and there were also significant differences in wetting tensions. Measuring adherence based on three different types of roughness and wetting tensions of forceps made from the same metal (titanium) also demonstrated a significant difference in the cleaning cycle. Histological examination of an artery coagulated with the gold-plated bipolar forceps showed that the structure had been completely collapsed without destruction of the layers, whereas arteries coagulated with the other materials revealed severely damaged structures. Adherence to bipolar forceps was dependent on both the material in the tips and the roughness of this material. The gold-plated bipolar forceps demonstrated the best performance. PMID- 14743927 TI - Cerebrovascular stereolithographic biomodeling for aneurysm surgery. Technical note. AB - Stereolithographic (SL) biomodeling is a new technology that allows three dimensional (3D) imaging data to be used in the manufacture of accurate solid plastic replicas of anatomical structures. The authors describe their experience with a patient series in which this relatively new visualization method was used in surgery for cerebral aneurysms. Using the rapid prototyping technology of stereolithography, 13 solid anatomical biomodels of cerebral aneurysms with parent and surrounding vessels were manufactured based on 3D computerized tomography scans (three cases) or 3D rotational angiography (10 cases). The biomodels were used for diagnosis, operative planning, surgical simulation, instruction for less experienced neurosurgeons, and patient education. The correspondence between the biomodel and the intraoperative findings was verified in every case by comparison with the intraoperative video. The utility of the biomodels was judged by three experienced and two less experienced neurosurgeons specializing in microsurgery. A prospective comparison of SL biomodels with intraoperative findings proved that the biomodels replicated the anatomical structures precisely. Even the first models, which were rather rough, corresponded to the intraoperative findings. Advances in imaging resolution and postprocessing methods helped overcome the initial limitations of the image threshold. The major advantage of this technology is that the surgeon can closely study complex cerebrovascular anatomy from any perspective by using a haptic, "real reality" biomodel, which can be held, allowing simulation of intraoperative situations and anticipation of surgical challenges. One drawback of SL biomodeling is the time it takes for the model to be manufactured and delivered. Another is that the synthetic resin of the biomodel is too rigid to use in dissecting exercises. Further development and refinement of the method is necessary before the model can demonstrate a mural thrombus or calcification or the relationship of the aneurysm to nonvascular structures. This series of 3D SL biomodels demonstrates the feasibility and clinical utility of this new visualization medium for cerebrovascular surgery. This medium, which elicits the intuitive imagination of the surgeon, can be effectively added to conventional imaging techniques. Overcoming the present limitations posed by material properties, visualization of intramural particularities, and representation of the relationship of the lesion to parenchymal and skeletal structures are the focus in an ongoing trial. PMID- 14743928 TI - Direct repair of a blisterlike aneurysm on the internal carotid artery with vascular closure staple clips. Technical note. AB - Vascular closure staple clips made of titanium were originally developed for microvascular anastomosis. Clinical applications for these clips include arteriotomy closure for carotid endarterectomy, extracranial-intracranial bypass, and dural closure. This is the first report in which vascular closure staple clips have been used successfully for direct repair of a tear on the internal carotid artery (ICA). This report involves a 65-year-old man who presented with sudden onset of headache. Admission computerized tomography scans demonstrated a diffuse and thick subarachnoid hemorrhage in the basal cisterns. Cerebral angiograms demonstrated a broad-based, small bulge on the superomedial wall of the left ICA. Intraoperatively, an extremely thin-walled aneurysm was seen on the segment of the ICA at the C-2 vertebral level. The aneurysm ruptured abruptly, although no surgical manipulation was being performed on the aneurysm itself. After temporary clips were applied on the vessel, a large tear of the ICA was repaired with vascular closure staple clips. Reconstruction with the vascular closure staple clips required only a short period of temporary occlusion of the ICA. Postoperative angiograms revealed reduction of the aneurysm bulge and good patency of the ICA. The postoperative course was uneventful, and the patient has been free of symptoms. The vascular closure staple clipping procedure is useful for urgent repair of an aneurysm tear. This method is a new treatment option for these fragile aneurysms in cases in which other options, such as encircling clips or bypass procedures, may have drawbacks or be impossible. PMID- 14743929 TI - Double-stent method: therapeutic alternative for small wide-necked aneurysms. Technical note. AB - The authors present two cases of patients with small, acutely ruptured, wide necked aneurysms of the distal vertebral artery that were not amenable to conventional coil embolization and were instead treated by means of a double stent method in which one stent was placed inside another. Angiography performed immediately after the procedure revealed a significant reduction in aneurysm filling; total occlusion of the lesion was observed after 7 days and confirmed 6 months later in both aneurysms. By placing one stent inside the other, stent permeability can be reduced, which may result in significant hemodynamic changes with accelerated aneurysm thrombosis. This double-stent method may represent a therapeutic alternative, especially in cases of small, wide-necked aneurysms in which conventional endovascular techniques or stent-supported coil embolization is not considered feasible or is believed to be too dangerous, and surgical treatment is contra-indicated. PMID- 14743930 TI - Transcranial electrical stimulation through screw electrodes for intraoperative monitoring of motor evoked potentials. Technical note. AB - The feasibility of high-frequency transcranial electrical stimulation (TES) through screw electrodes placed in the skull was investigated for use in intraoperative monitoring of the motor pathways in patients who are in a state of general anesthesia during cerebral and spinal operations. Motor evoked potentials (MEPs) were elicited by TES with a train of five square-wave pulses (duration 400 microsec, intensity < or = 200 mA, frequency 500 Hz) delivered through metal screw electrodes placed in the outer table of the skull over the primary motor cortex in 42 patients. Myogenic MEPs to anodal stimulation were recorded from the abductor pollicis brevis (APB) and tibialis anterior (TA) muscles. The mean threshold stimulation intensity was 48 +/- 17 mA for the APB muscles, and 112 +/- 35 mA for the TA muscles. The electrodes were firmly fixed at the site and were not dislodged by surgical manipulation throughout the operation. No adverse reactions attributable to the TES were observed. Passing current through the screw electrodes stimulates the motor cortex more effectively than conventional methods of TES. The method is safe and inexpensive, and it is convenient for intraoperative monitoring of motor pathways. PMID- 14743931 TI - Novel closure technique for the endonasal transsphenoidal approach. Technical note. AB - Transsphenoidal microsurgery has been the standard approach to sellar lesions since the repopularization of the technique with modifications by Dott, Guiot, and Hardy. The endonasal transseptal transsphenoidal approach, as introduced by Hirsch, is still commonly used by pituitary surgeons to remove lesions of the sellar and parasellar region. One disadvantage of this approach is that the submucosal dissection requires postoperative nasal packing, which is a source of discomfort in patients who undergo transsphenoidal surgery. The authors describe a novel closure technique for the unilateral endonasal transsphenoidal approach that eliminates the need for full nasal packing, minimizing postoperative rhinological morbidity. This technique has been performed in 67 patients harboring sellar and parasellar lesions. All patients recovered rapidly without significant rhinological sequelae. PMID- 14743932 TI - Guyon canal. PMID- 14743933 TI - Subdural hematoma. PMID- 14743934 TI - Lamina terminalis fenestration. PMID- 14743935 TI - Spasmodic torticollis. PMID- 14743936 TI - Risk factors that predicted problem drinking in Danish men at age thirty. AB - OBJECTIVE: The Danish Longitudinal Study of Alcoholism utilized a prospective, high-risk research paradigm to identify putative markers of adult male alcoholism from a comprehensive database that began with the birth of the subject and extended over three decades. This article focuses on measures antedating abusive drinking that predicted lifetime alcohol abuse/dependence at age 30 years. METHOD: The original 330 subjects of this study were drawn from a large Danish birth cohort (N = 9,125) born between 1959 and 1961. The sample included 223 sons of treated alcoholic fathers (high-risk group) and 107 matched sons whose biological fathers had no record of treatment for alcoholism (low-risk group). This sample has been thoroughly investigated with a variety of methods representing multiple domains that included perinatal records, pediatric records, school records, teacher ratings, school physician records and a series of structured interviews and psychometric tests at ages 19-20 and 30 years. The present analysis focuses on the degree to which premorbid differences between the high- and low-risk groups later predicted lifetime drinking problems at age 30 (n = 241). RESULTS: As expected lifetime alcohol abuse/dependence by age 30 was reported significantly more often in the high-risk group. Of the 394 premorbid variables tested, 68 were found to distinguish the high- from the low-risk group before any subjects had developed a drinking problem. Of these 68 variables, 28 (41%) were also associated with DSM-III-R alcohol abuse/dependence at age 30. These 28 putative markers were reduced to 12 that were entered into a multiple regression analysis to search for the most powerful unique predictors of alcoholism. Four of the 28 putative markers were independently associated with problem drinking at age 30: low birth weight, number of life crises in childhood, ratings of childhood unhappiness and antisocial personality disorder. The regression model accounted for 46% of the drinking outcome variance. A father's alcoholism by itself no longer independently contributed to the prediction of his son's drinking and with one exception, did not systematically interact with the putative markers to facilitate the prediction of alcohol dependence at age 30. CONCLUSIONS: Risk itself. which significantly predicted problem drinking at age 30, was not uniquely associated with the development of alcoholism in adulthood. These findings, rather, provide broad support for the biopsychosocial model of alcoholism, especially for those models that emphasize the cumulative influence over time of internal and external variables in biologically vulnerable individuals. PMID- 14743937 TI - The role of women's substance use in vulnerability to forcible and incapacitated rape. AB - OBJECTIVE: Although women's substance use is believed to contribute to rape vulnerability, few studies have examined, separately from forcible rape, rape that occurs due to incapacitation. The current study examines the prevalence of completed forcible and incapacitated rape in a representative community sample of young women, identifies the predictors of these two forms of rape and compares characteristics of forcible and incapacitated rape incidents. METHOD: Women, ages 18-30 (N = 1,014), were recruited from households in the Buffalo, NY, area, by means of random-digit-dialing. They completed computer-assisted measures, including the Sexual Experiences Survey and a face-to-face interview regarding sexual assault experiences occurring since age 14. RESULTS: Lifetime prevalence of incapacitated rape was nearly identical to prevalence of forcible rape, with about 1 in 10 women reporting each type of rape since age 14. In multivariate analysis, age and childhood sexual abuse predicted forcible but not incapacitated rape, whereas adolescent alcohol and drug use predicted incapacitated but not forcible rape. Incapacitated rape incidents differed from forcible rape incidents on several contextual variables, including relationship to perpetrator, activities preceding the assault and victim injury. CONCLUSIONS: Findings suggest that forcible rape and incapacitated rape may be different forms of sexual assault, with different distal and proximal correlates. Distinguishing these two forms may facilitate understanding of the role of women's alcohol and drug use in sexual assault. PMID- 14743938 TI - Characteristics of violent bars and bar patrons. AB - OBJECTIVE: The present analysis is an attempt to examine the characteristics of bars in which violence occurs while accounting for the personalities of the clientele that frequent the bar. It is proposed that an explanation of why violence occurs at certain bars requires examining the characteristics of the bars, the personalities of the clientele, and how these two types of variables act together in order to give rise to aggressive behavior. METHOD: We conducted interviews with frequent bar patrons (n = 327), assessing participants on a number of individual differences related to aggression and drinking behavior as well as on characteristics of the usual bar that they attend. Bars were categorized into violent bars (n = 256) or nonviolent bars (n = 71) based on participant responses. RESULTS: Participants' age, alcohol dependence and anger expression differentiated those who frequented violent bars from those who frequented nonviolent bars. The relationship of these individual differences to bar type was mediated by a number of characteristics of the bar itself, including noise, temperature, the presence of bouncers, the gender of the workers, the presence of billiards and illegal activities in the bar. CONCLUSIONS: The results indicate that individuals having certain personality characteristics are attracted to bar environments that promote antinormative behaviors such as violence. However, it seems to be the characteristics of the bars that are the strongest predictors of violence. PMID- 14743939 TI - Alcohol use in pregnant low-income women. AB - OBJECTIVE: The present study had two aims: (1) to examine the prevalence rates of prenatal alcohol consumption in a group of women participating in the Special Supplemental Nutrition Program for Women, Infants and Children (WIC) in Southern California, with special emphasis on Hispanic women, and (2) to identify variables associated with postconception drinking in low-income minority women. METHOD: The study employed a cross-sectional survey that assessed alcohol use rates, demographic variables, and alcohol risk status. Participants were 826 pregnant women enrolled in the Public Health Foundation Enterprises (PHFE) WIC Program in Los Angeles and Orange counties, California. Pregnant women who enrolled self-administered a specially designed alcohol screener. RESULTS: Findings were that 24% of sample women were consuming alcohol post conception. Of that percentage, approximately two thirds drank prior to pregnancy recognition, and one third continued to drink after pregnancy was confirmed. Approximately 30% of white non-Hispanic, black non-Hispanic and English-speaking Hispanic women were found to drink post conception compared with 15.8% of Spanish-speaking Hispanic women. This finding supported recent research that suggests more acculturated Hispanic women tend to incorporate the drinking patterns of the larger U.S. population to a greater extent than less acculturated Hispanic women. Although a number of demographic variables differentiated women who were abstinent from those who were postconception drinkers, the best predictor of postconception alcohol consumption was the woman's high-risk drinking score as measured by the TWEAK (sensitivity = 70.1%, specificity = 88.5%). CONCLUSIONS: Results suggest the importance of screening low-income minority pregnant women in a community setting so that interventions can be initiated to prevent fetal alcohol syndrome and related conditions. PMID- 14743940 TI - Alcohol consumption and marital status of French women in the GAZEL cohort: a longitudinal analysis between 1992 and 1996. AB - OBJECTIVE: The purpose of this research was to describe women's consumption of alcoholic beverages as it related to their marital status over a 5-year period (1992 through 1996) and to study alcohol consumption around the time of marriage or divorce. METHOD: The study sample comprised 4,782 women who worked for Electricite de France-Gaz de France and belonged to the GAZEL cohort. The relevant variables collected by five successive annual questionnaires included marital status and alcohol consumption characteristics. Marginal models were used, reflecting the fact that the data were not independent. RESULTS: Divorcees and widows drank less than married women as measured in fewer glasses per day and fewer days per week drinking wine. Women in the oldest generation drank more than the younger women. Getting married was accompanied by an increased level of drinking, especially of wine, beginning a year before the wedding and lasting until 4 years after it. Consumption declined briefly during the year after a divorce. CONCLUSIONS: These results may be useful for designing prevention programs aimed at groups of women in the general population in France. PMID- 14743941 TI - Gender comparison of alcohol exposure on drinking occasions. AB - OBJECTIVE: This study was designed to assess the influence of gender, age, drinking style and other selected demographic variables on the rate of alcohol beverage consumption on drinking days. The data were used to predict possible differences in ethanol exposure levels (peak blood alcohol concentration [BAC]) among the subgroups examined. METHOD: A representative sample (N = 2,627) of the U.S. adult population was surveyed using computer-assisted telephone interviewing to assess alcohol use (past 12 months) in quantity, frequency and type of beverage, as well as the time (minutes) usually required to consume the typical quantity of alcohol on drinking days. RESULTS: Of the 1,833 current drinkers (1,028 men, 805 women), women reported consuming a mean of 2.2 standard drinks (1 drink = 12 g ethanol) on typical drinking occasions (days); men reported consuming a mean of 3.2. The duration of the drinking episode was similar for women (122 minutes) and men (126 minutes). The hourly rate of drink consumption was thus lower for women (approximately 1.1 drinks/hour) than for men (approximately 1.6 drinks/hour). For both men and women, there was an age-related increase in the frequency of drinking but a decrease with age in predicted peak BACs and in the alcohol intake per drinking episode. Peak BACs achieved during typical drinking episodes were estimated to be quite similar for men (0.037) and for women (0.036) when prediction equations were based on equal rates of alcohol elimination in both genders. The estimated peak BACs were lower in women than in men when the prediction equation assumed a higher rate of ethanol elimination in women. CONCLUSIONS: Information on the rate of alcohol beverage consumption on drinking days facilitates prediction of BACs typically experienced in men and women of different demographic group memberships. The exposure values obtained will depend upon as yet unsettled norms for ethanol elimination rates in men and women. PMID- 14743942 TI - Ethanol-induced advance in the onset of puberty is prevented by the GABAA antagonist bicuculline in female rats. AB - OBJECTIVE: The use of alcohol during lactation produces an advance in the onset of puberty in female rats. This research studied the possible participation of GABAA activity in this phenomenon. METHOD: Female Wistar rats were exposed to one of three different treatments from 13 to 18 days of postnatal age: (1) ethanol (2.5 g/kg PO) twice a day and isotonic saline (10 ml/kg IP) four times a day (AS group), (2) ethanol (2.5 g/kg PO) twice a day and bicuculline (2.5 mg/kg IP) four times a day (ABic group), (3) water (25 ml/kg PO) twice a day and bicuculline (2.5 mg/kg IP) four times a day (WBic group). A fourth group (control) did not receive any drug. RESULTS: The advance in the age at vaginal opening and at the first vaginal estrous seen in females of the AS group was completely prevented in the ABic group when bicuculline was added to alcohol treatment. The age at first behavioral estrous, however, was not different between groups. The age at vaginal opening, first vaginal estrous and first behavioral estrous was not significantly different in control, ABic and WBic groups. Body weight at vaginal opening was lower in the AS group than in the other three groups. Gestational period was longer in the AS group, but the number of pups per litter was lower in females of the ABic and WBic groups. CONCLUSIONS: The results reported here support a significant role for GABAA activity in the effects of alcohol at the onset of puberty. PMID- 14743943 TI - Actions of ethanol on epidermal growth factor receptor activated luteinizing hormone secretion. AB - OBJECTIVE: Activation of the epidermal growth factor receptor (EGF-R) stimulates prepubertal luteinizing hormone-releasing hormone (LHRH) release, which in turn induces luteinizing hormone (LH) secretion. Although ethanol (ETOH) diminishes LHRH secretion and delays the onset of female puberty, its actions following EGF R activation are unknown. We therefore investigated the effects of ETOH on EGF induced LHRH and LH release, both in vivo and in vitro. METHOD: Basal blood samples were taken every 15 minutes from immature female rats, which then received ETOH (3 g/kg) or saline by gastric gavage. After a 90-minute ETOH absorption period, a single blood sample was drawn from each rat. Finally, EGF (200 ng/3 microl) was injected into the third ventricle of all animals and postinjection samples were drawn every 15 minutes. A similar experimental design was performed except no blood samples were taken. After ETOH exposure, EGF was administered, and 45 minutes later the animals were killed and their medial basal hypothalamus and pituitary were removed and analyzed for EGF-R and cyclooxygenase (COX) 1 and 2 by western blot analysis. For in vitro experiment, median eminence fragments from immature female rats were exposed to ETOH in a static incubation system, and prostaglandin-E, (PGE2) and LHRH were measured. RESULTS: EGF stimulated LH release, and this release was blunted (p < .05) by ETOH. Western blot analysis revealed that ETOH did not alter the EGF-R protein levels in the hypothalamus. ETOH blocked EGF-induced PGE2 and LHRH released from isolated median eminences. EGF administration increased both COX-1 (p < .001) and COX-2 (p < .01), but both enzymes were blocked by ETOH. CONCLUSIONS: The ETOH-induced decrease in EGF-stimulated LH release is due to a reduction in the formation of hypothalamic PGE2 and, subsequently, suppressed LHRH release. PMID- 14743944 TI - Drinking to extremes: theoretical and empiricalanalyses of peak drinking levels among college students. AB - OBJECTIVE: Heavy drinking among college students continues to be a substantial problem on campuses across the United States. Attempts to predict these drinking events have been restricted to assessments of the correlates of heavy drinking (measured at 4 or 5 drinks) and have not examined the peak drinking levels that can be fatal to students. This article presents a theoretical analysis of college drinking patterns that provides a basis for estimating peak drinking levels and predicts future risks related to peak drinking events. METHOD: Survey data were collected on sociodemographics and drinking patterns of 2,102 college students from two college campuses in California. A mathematical model of drinking patterns was used to characterize the stochastic distribution of drinking events among 1,273 students who drank five or more times and consumed more than one drink on some occasion since the beginning of the school year. An application of extreme value theory enabled the estimation of peak drinking levels for every college drinker. These estimates were related to self-reported maximum drinking levels and sociodemographic characteristics of respondents. RESULTS: Among these drinkers, the distribution of self-reported maximum drinking levels ranged from 2 to 43 drinks per occasion. Estimated peak drinking levels ranged from 3 to 49. Maximum drinking levels were well characterized by peak drinking estimates (R2 = 0.503). Variations in peak drinking levels were large and specifically related to particular sociodemographic groups (i.e., white male freshmen). CONCLUSIONS: The theoretical model of peak drinking events effectively characterizes maximum drinking levels among college students. High levels of peak drinking are to be expected among specific sociodemographic subgroups. These risks can be assessed on an individual basis. At the population level, risks for harm related to peak drinking events are predictable. PMID- 14743945 TI - Negative consequences of intercollegiate athlete drinking: the role of drinking motives. AB - OBJECTIVE: Intercollegiate athletes consume more alcohol and experience more negative alcohol-related consequences than nonathletes. The purpose of this study was to determine if drinking motives accounted for variability among intercollegiate athletes in experiencing negative alcohol-related consequences, and to analyze how the strength and patterns of the relationship between individual drinking motives and negative consequences varied among the different consequences. METHOD: Self-report data were analyzed on 206 National Collegiate Athletic Association Division I athletes (55% female), selected from a large, midwestern university, who reported drinking alcohol in the past year. Participants completed the Drinking Motives Measure and the Core Alcohol and Drug Survey. RESULTS: Drinking motives accounted for variability in experiencing negative alcohol-related consequences. Drinking for coping reasons displayed the strongest relationship with most of the negative consequences, but for some consequences social and enhancement motives displayed relationships with the consequences that were either similar to or stronger than those of the coping motives. CONCLUSIONS: This research demonstrated that drinking motives in general are useful predictors of negative alcohol-related consequences among a yet unstudied population, intercollegiate athletes. The findings suggest that drinking for negatively reinforcing reasons (i.e., coping motives) is generally the strongest motivational predictor of alcohol-related consequences, although the relative strength of individual motives in predicting consequences can vary depending upon the content of an individual consequence. PMID- 14743946 TI - Perceived social norms and their relation to university student drinking. AB - OBJECTIVE: The aims of this study were to compare university students' perceptions of drinking norms with actual student drinking norms, to examine the relationship between norm misperception and individual drinking status and to investigate the relative importance of three reference groups as potential determinants of individual drinking levels: young people in New Zealand of the same age and gender, local university students of the same age and gender and the closest friends of individual respondents. METHOD: In 2002 a randomly selected representative sample of 1,564 New Zealand university students aged 16-29 years completed an Internet-based survey of their alcohol use (response rate: 82%). Respondents were asked to estimate the incidence of heavy episodic drinking and vomiting in the three reference groups and to rate their own drinking in comparison. Estimates within +/- 10% of actual norms were rated as accurate; estimates above or below actual norms by more than 10% were rated, respectively, as overestimates and underestimates. RESULTS: The vast majority of women (80%) and men (73%) overestimated the incidence of heavy drinking among student peers. The incidence of vomiting was also overestimated, but to a lesser extent. The extents of overestimation for both heavy drinking and vomiting were strongly related to the individual's heavy drinking frequency (p < .001). Only 9% of drinkers believed they drank more than other students. Correlations of perceived norms and self-reported drinking increased with the proximity of the social grouping to the individual and were higher for women than for men. CONCLUSIONS: This New Zealand university sample showed strong evidence of norm misperceptions, consistent with the results of several U.S. studies. Perceived norms are strongly related to individual drinking levels. It is unclear whether norm misperceptions are a cause or effect of heavy drinking. Research in which norm misperceptions are corrected may assist in understanding their importance in the etiology and treatment of heavy drinking. PMID- 14743947 TI - Malt liquor use, heavy/problem drinking and other problem behaviors in a sample of community college students. AB - OBJECTIVE: This study examined the prevalence of malt liquor use and its relationship with heavy or problem drinking, other substance use and other problem behaviors in a sample of 1,029 (58% female) students attending a community college in California. METHOD: Data were collected using anonymous surveys that were administered during daytime classes. Bivariate analyses were conducted to examine relationships between malt liquor use and demographic characteristics, heavy/problem drinking and other types of substance use and problem behaviors. Logistic regression analyses also were conducted to determine whether malt liquor use was predictive of problem drinking, drug use and other problem behavior after adjusting for background variables. RESULTS: Malt liquor use was most prevalent among male, white and Latino students. Compared with nonmalt liquor drinkers, malt liquor drinkers consumed significantly more alcohol, engaged in heavy drinking more often and experienced intoxication more often. Malt liquor drinkers scored significantly higher on the Alcohol Use Disorders Identification Test than nonmalt liquor drinkers. Malt liquor drinkers were more likely than nonmalt liquor drinkers and abstainers to report use of cigarettes, marijuana and illicit drugs and to exhibit problem behaviors. Malt liquor use was predictive of problem drinking, drug use and other problem behavior after adjusting for demographics and alcohol use in regression analyses. CONCLUSIONS: The findings of this study suggest that malt liquor use is associated with heavy and problem drinking, other drug use and behavioral problems among community college students. PMID- 14743949 TI - The costs of screening and brief intervention for risky alcohol use. AB - OBJECTIVE: The purpose of this study was to estimate provider-incurred costs of alcohol screening and brief intervention (SBI) for risky drinking as implemented in four managed care organizations (MCOs) participating in the Cutting Back project implemented by the University of Connecticut Health Center. METHOD: Each MCO provided two comparable primary care clinics in which two different SBI models were implemented: the "Practitioner" (P) model and the "Specialist" (S) model. Risky drinkers were identified based on responses to a health appraisal form. They were administered the AUDIT to determine an appropriate intervention. Using data collected from these sites, we separately estimated start-up and ongoing implementation costs of the intervention. RESULTS: SBI start-up costs per MCO ranged from approximately dollars 86,000 to dollars 115,000 across the four study MCOs. Across all four study MCOs, the estimated median ongoing implementation cost of administering the health appraisal was dollars 0.25 per patient appraised, and the estimated median cost of screenings was dollars 0.42 per patient screened. The estimated median cost of performing the brief intervention across the study MCOs was dollars 2.59 per patient receiving the intervention in the S clinics and dollars 3.43 per patient receiving the intervention in the P clinics. Labor costs dominated start-up and ongoing implementation. Technical assistance costs accounted for a significant proportion of start-up costs. Implementation in the S model is less costly than in the P model, largely because of the S model's use of less expensive nonphysician labor. CONCLUSIONS: Our analysis suggests that the cost of SBI is modest, and MCOs may want to consider adopting SBI as an alcohol use prevention tool. Although our results suggest that the S model is less costly than the P model, clinic-level implementation factors may affect the relative costs of the S versus P models. PMID- 14743948 TI - The influence of race and religion on abstinence from alcohol, cigarettes and marijuana among adolescents. AB - OBJECTIVE: Past research has not fully explained why black youth are less likely than white youth to use alcohol and other substances. One plausible yet underexamined explanation is the "religion hypothesis," which posits that black youth are more likely than white youth to abstain because they are more religious than white youth. The present study tested this hypothesis empirically. METHOD: The study examined data from large, nationally representative samples of white and black 10th graders from the Monitoring the Future project. RESULTS: Relative to white students, black students are more likely to abstain from alcohol, cigarettes and marijuana and are more highly religious. Consistent with the "religiosity hypothesis," race differences in abstinence are substantially reduced when race differences in religiosity are controlled. Unexpectedly, however, highly religious white youth are more likely than highly religious black youth to abstain from alcohol and marijuana use. CONCLUSIONS: Although religion is an important protective factor against alcohol and other substance use for both white and black adolescents, it appears to impact white youth at an individual level, whereas for black youth the influence of religion seems greatest at the group level. Future research should seek to better understand the mechanisms through which religion promotes adolescents' abstinence from the use of drugs and should seek to explain why the magnitude of its effect varies for black and white adolescents. PMID- 14743950 TI - Comparison of a quick drinking screen with the timeline followback for individuals with alcohol problems. AB - OBJECTIVE: Two major strategies have typically been used to assess recent drinking: (1) Daily Estimation (DE) measures such as the Timeline Followback (TLFB) and (2) Quantity-Frequency (QF) summary measures. Although QF measures provide a quick and easy measure of consumption, they have been criticized as not being able to capture sporadic and unpatterned drinking (e.g., days that reflect important social and/or health risks). The TLFB, a psychometrically sound drinking assessment method, is able to capture all drinking, including sporadic heavy days and unpatterned drinking. In some situations, however, recall of daily drinking may not be possible or practical (e.g., limited time; no resources). This article compares results obtained by using a QF measure and a DE measure to assess problem drinkers' pretreatment drinking. METHOD: The current study, part of a large community mail intervention with 825 alcohol abusers, compared results from two drinking measures covering the same time interval that were administered on two different occasions approximately 2.5 weeks apart. Both measures, the Quick Drinking Screen (QDS; a QF summary measure that collected data by telephone) and the TLFB (a self-administered daily estimation measure), collected drinking data for the year prior to the interview. RESULTS: Although the QDS and the TLFB are very different drinking measures, remarkably similar aggregate drinking data were obtained for five drinking variables. CONCLUSIONS: When it is not necessary or possible to gather detailed drinking data, the QDS produces reliable brief summary measures of drinking, at least for not severely alcohol dependent individuals. Also, respondents do not appear to use a repetitive response pattern when completing the TLFB. PMID- 14743951 TI - The role of community services and informal support on five-year drinking trajectories of alcohol dependent and problem drinkers. AB - OBJECTIVE: The purpose of this study was to examine how informal support and community services impact the course of alcohol disorders by examining the trajectories of alcohol consumption over 5 years in dependent and problem drinkers. METHOD: Alcohol dependent adults (n = 600) and problem drinkers (n = 992) were identified through probability surveys in the general population and in public and private treatment programs throughout a California county. Participants were interviewed at baseline and again 1, 3 and 5 years later. Models controlling for demographic characteristics, problem severity, community services and recovery-oriented social networks were estimated, using a multi level, mixed model to predict alcohol consumption over time. RESULTS: A significant trend of reduced drinking over time was observed for both dependent and problem drinkers. Recovery-oriented social networks and AA participation predicted decreased consumption for both groups. Contacts with medical, mental health, welfare and legal systems were predictive of reduced consumption for problem drinkers. In the dependent group, only contacts with mental health agencies marginally predicted decreased consumption. CONCLUSIONS: Findings point to the importance of developing mechanisms for better identifying problem drinkers in the course of contacts with health and social service systems and for facilitating use of self-help groups and positive changes in social networks. Development of recovery-oriented social networks should be emphasized to extend the benefits of treatment for dependent individuals. PMID- 14743952 TI - Positive and negative effects of social support on the relationship between work stress and alcohol consumption. AB - OBJECTIVE: This study was performed to evaluate both positive and negative influences of social support on the relationship between work stress and alcohol consumption in male white-collar workers. METHOD: This study was performed in a population of male white-collar workers in Osaka, Japan. The subjects (N = 661) were categorized into nine subgroups based on the moderated mediation model of Frone and on the results of signal detection analysis using chi-square parameters calculated from work stressors and depression (mediators). Hierarchical linear regression analyses were performed among the nine subgroups to test the effects of interactions between four types of social support (i.e., two types of belonging support and two types of appraisal support) and work stress on alcohol consumption. RESULTS: Our results verified that social support has both positive and negative effects on the relationship between work stress and alcohol consumption: (1) interactions between "belonging support (1)" and depressive symptoms were related to decreased alcohol consumption in two subgroups (p < .01 in Group 1 and p < .05 in Group 6), whereas the interaction was related to increased alcohol consumption in one subgroup (p < .01 in Group 7); (2) interaction between "belonging support (2)" and depressive symptoms was related to decreased alcohol consumption in two groups (p < .05 in Group 5 andp < .01 in Group 7), whereas the interaction was related to increased alcohol consumption in one group (p < .01 in Group 1). CONCLUSIONS: Social support was suggested to have various types of influences (i.e., positive, negative or no effect) on the relationship between work stress and alcohol consumption, depending on the type of social support and environmental factors defining the groups. PMID- 14743953 TI - Alcohol consumption by elderly Americans. AB - OBJECTIVE: The purpose of this study was to estimate the prevalence of alcohol consumption in Americans age 65 years and older using data from three nationally representative cross-sectional surveys: the National Health Interview Survey (NHIS-2000), the Behavioral Risk Factor Surveillance System (BRFSS-2001) and the National Household Survey on Drug Abuse (NHSDA-2000). METHOD: Alcohol consumption levels were defined as none, moderate (< or = 1 drink a day) and heavier (> 1 drink a day). The NHIS assessed alcohol consumption in the past year, and the BRFSS and NHSDA assessed alcohol consumption in the past 30 days. Differences between the BRFSS and NHSDA were tested using multinomial logistic regression. Age trends in alcohol consumption (between age 65 and 84 years) were tested using logistic regression. All analyses were weighted to produce national estimates. RESULTS: In men, the prevalence of moderate drinking was 37.6% (95% confidence interval [CI]: 35.2-40.0) in the NHIS, 38.7% (CI: 37.3-40.1) in the BRFSS and 27.2% (CI: 23.6-30.8) in the NHSDA. The prevalence of heavier drinking among men was 10.1% (CI: 8.7-11.5), 10.1% (CI: 9.3-10.9) and 9.2% (CI: 7.2-11.3), respectively. In women, the prevalence of moderate drinking was 32.3% (CI: 30.4 34.2) in the NHIS, 27.7% (CI: 26.7-28.6) in the BRFSS and 21.5% (CI: 18.9-24.2) in the NHSDA. The prevalence of heavier drinking among women was 2.2% (CI: 1.6 2.7), 2.6% (CI: 2.3-2.9) and 2.4% (CI: 1.4-3.3), respectively. In increasingly older groups of men, moderate drinking remained stable (all surveys, p for age trend [p trend] = NS), while heavier drinking significantly decreased in two of three surveys (NHIS and BRFSS, p trend < .05; NHSDA, p = NS). Conversely, in increasingly older groups of women, moderate drinking significantly decreased (all surveys, p trend = .001), while heavier drinking remained stable (all surveys, p trend = NS). CONCLUSIONS: In the years 2000 to 2001 approximately one third of the U.S. elderly population, about 11 million persons, consumed alcohol. The risks and benefits of drinking by elderly Americans will become an increasingly important public health issue as this segment of the population expands over the coming decades. PMID- 14743954 TI - Association of biochemical values with morbidity in the elderly: a population based Swedish study of persons aged 82 or more years. AB - BACKGROUND: Various inter-dependent factors influence serum biochemical values. In the elderly, the impact of these factors may differ compared with younger age groups and therefore population-based studies among older people are needed. The specific morbidity in old age, including also various types of drug therapy, should be observed. METHODS: Various biochemical tests in 349 females and 186 males over 81 years of age were carried out and the associations of biochemical values with morbidity, drug therapy, anthropometry and gender were estimated. RESULTS: Biochemical serum values deviate in various diseases, characterized by increased frequency in the elderly, i.e. congestive heart failure, osteoporosis, hip fractures, depression and dementia. All of these diseases present a tendency to increased homocysteine, usually combined with low folate. Cases with intact cognitive function throughout the six years after sampling are characterized by low homocysteine, which is the opposite of what is found in dementia. Furthermore, congestive heart failure is associated with impaired creatinine clearance and increased urea and urate, and osteoporosis and hip fractures are characterized by low albumin and cholesterol. Increased values for urate and impaired creatinine clearance are found in coronary diseases. In gout, multiple biochemical changes take place. For cases with a history of diabetes, arterial hypertension, peptic ulcer and malignancy, few changes are found compared with the values of the total sample. Furosemide therapy is associated with the same pattern as congestive heart failure, and laxative treatment is characterized by low folate and high homocysteine values. PMID- 14743955 TI - Simultaneous genotyping of alcohol dehydrogenase 2 and aldehyde dehydrogenase 2 by single-strand conformation polymorphism analysis. AB - Alcohol dehydrogenase (ADH; EC 1.1.1.1) and aldehyde dehydrogenase (ALDH; EC 1.2.1.3) have important roles in the elimination of ingested ethanol. These enzymes have polymorphisms resulting from single-point mutations that cause kinetic differences in their respective enzyme activities. Simultaneous observation of these enzymes would be useful in investigating the association between these enzyme polymorphisms and alcohol-related problems. In this study amplified genomic DNA was amplified from nail clippings with two sets of primers for ADH2 and ALDH2 genes, respectively, in a micro test tube and the accuracy of the amplification was verified by direct sequencing. The PCR products were separated into four distinct bands by single-strand conformation polymorphism analysis. This genotyping method is fast, accurate. reliable and inexpensive, and requires the same amount of template DNA as non-simultaneous methods. In other words, the required amount of template DNA for this method is only half that required for the separate genotyping of ADH2 and ALDH2. PMID- 14743956 TI - Pulmonary function in rats dying from long-term parenteral nutrition. AB - Infusion of Vamin or Intralipid causes death in a rat model of continuous parenteral nutrition. Morphological investigations have shown vascular injury and thrombus formation in the lungs. In this study, lung function in rats was examined before death due to parenteral nutrition. The rats were fed saline intravenously (group I); 100 mL kg(-1) day(-1) (controls); a 7% amino acid glucose solution (Vamin-Glukos) (group II); 100 mL kg(-1) day(-1), or 20% fat emulsion (Intralipid) (group III); 40 mL kg(-1) day(-1). The infusion was stopped when the condition of the rats deteriorated. In a saline-perfused, isolated lung model, pulmonary arterial pressure (Ppa), transpulmonary pressure (Ptp), endothelial function, measured as inactivation of serotonin (bioassay), and the capillary filtration coefficient (CFC) were determined. Haematological parameters were also evaluated. Constant findings in group II and III were central thrombus formation, anaemia and thrombocytopenia. Ppa increased from 0.7 (0.04) kPa in group I to 1.4 (0.1) kPa and 1.7 (0.1) kPa in groups II and III, respectively (p<0.001). Inactivation of serotonin was reduced to 36% (2) in group II and 37% (2) in group III compared with 74% (5) in group I (p<0.002). CFC increased to 25 mg min(-1) (5) (group II) and 30 mg min(-1) (6) (group III) compared with 13 mg min(-1) (2) in controls (p=0.01). The study shows that major pulmonary hypertension and severe reduction of the endothelial function are present when rats deteriorate after infusion of parenteral nutrition substrates. PMID- 14743957 TI - Changes in the plasma levels of protein C system parameters in pregnancy. AB - Resistance to activated protein C (APC) is a condition that leads to a hypercoagulative state with an increased risk for venous thrombosis. The aim of this study was to test the functionality of the protein C system in normal and complicated pregnancies and APC resistance. A total of 131 patients were tested, including pregnant women with normal and complicated pregnancies at different periods, e.g. from weeks 1 to 20, 21 to 30 and 31 to 38 of gestation. The following hemostatic parameters were determined: protein S, protein C, protein C Global and protein C APC sensitivity. Commercial "Behring" tests were used to determine the parameters of hemostasis. The values for protein C activity were within normal limits. Protein S values were below the lower limits. Significantly lower PC-NR (protein C normalized ratio) and APC-NR (activated protein C normalized ratio) values were found in all three of the gestation periods in pregnant women with a history of repeated miscarriages and hypertension in relation to pregnant women with normal pregnancies. In order to assess the diagnostic accuracy of investigated hemostatic parameters as markers of prethrombotic changes in pregnant women, the obtained values of investigated hemostatic parameters were evaluated by ROC analysis. PC-NR and APC-NR showed satisfactory diagnostic accuracy as markers of prethrombotic changes in pregnant women: more precisely, they were found to be good indicators of resistance to activated protein C in pregnancy. PMID- 14743958 TI - Serum iron concentration as a tool to measure relative iron absorption from elemental iron powders in man. AB - OBJECTIVE: To present a new method for measuring the relative bioavailability (RBV) of commercial elemental iron powders by investigating induced changes of serum iron concentration (S-Fe) in relation to ferrous sulphate (FeSO4). Earlier studies have shown that in a group of subjects there is good agreement between the increase in S-Fe and the amount of iron absorbed from a simple iron salt as FeSO4. METHODS: The study included two groups of male blood donors (n=2 x 16 subjects) who were served three meals with an interval of approximately nine weeks between each one. In one group the meal was fortified with reduced iron, ferrous sulphate or no iron at all. In the other group the meal was fortified with electrolytic iron, ferrous sulphate or no iron. The S-Fe increase was followed for 6 h. Studying the non-iron meals was necessary so that the basal diurnal variation in the S-Fe during the six hours could be measured and subtracted. RESULTS: The mean S-Fe increase calculated as the area under the curve (AUC) from the reduced iron (RBV=0.43) and the electrolytic iron (RBV=0.73) differed significantly from the AUC following FeSO4 (p=0.002 and p=0.021, respectively). The difference between the reduced and the electrolytic iron was also statistically significant (p=0.036). CONCLUSION: Measuring increases in S-Fe could be a reliable and simple method to determine the RBV in comparative studies of elemental iron powders in relation to FeSO4. PMID- 14743959 TI - Serum lysophosphatidic acid concentrations measured by dot immunogold filtration assay in patients with acute myocardial infarction. AB - In this study the relation between lysophosphatidic acid (LPA) and myocardial infarction was investigated, the typical and simplified methods for measuring serum LPA concentration by dot immunogold filtration assay (DIFA) based on a polyclonal antibody to LPA were developed, and serum LPA concentrations were measured in 31 patients with acute myocardial infarction (AMI) and 12 controls (blood donors) by DIFA. Serum LPA levels were raised more than twofold 8 h after the onset of AMI. Maximal elevation (10.43 mg/L) was found at 48-72 h following onset and remained higher than the control concentration (1.66 mg/L) 7 days after AMI. The rise in serum LPA concentration in AMI patients suggests that LPA might be involved in AMI-related pathophysiology in the cardiovascular system. The simplified DIFA developed in the present study for measuring serum LPA concentration is convenient and highly sensitive. PMID- 14743960 TI - Epidermal growth factor suppresses interferon-induced accumulation of p53 and p21 by using protein kinase C. AB - The tumor suppressor protein p53 is the most frequently mutated gene in human cancers. Since its discovery, p53 has evolved from a potential oncogene to the principal tumor suppressor in humans. p53 protects not only against oncogenic stress but also against the presence of DNA damage. Now, p53 is positioned at the vertex of cellular signals warning of threats of genomic damage and oxidative stress. Under these conditions p53 is phosphorylated by multiple kinases and these phosphorylations not only increase its half-life but also increase its localization in the nucleus. p53 localized in the nucleus induces cell-cycle arrest to allow repair processes or, failing that, promotes cellular senescence or cell death. In this study it is shown that treatment of ME180S cells with interferon alpha (IFN-alpha) and interferon gamma (IFN-gamma) result in time dependent accumulation of p53 and its transcriptional target, p21. Pretreatment of ME180S cells with epidermal growth factor (EGF) inhibits IFN-dependent induction of p53 and p21 by protein kinase C dependent pathways. PMID- 14743961 TI - Monocyte chemoattractant protein-1 and CC-chemokine receptor-2 in severe hypercholesterolaemia. AB - OBJECTIVES: To investigate whether plasma concentrations of monocyte chemoattractant protein-1 (MCP-1) and the gene expression of its receptor on the monocyte cell surface CCR-2 were elevated above normal in subjects with asymptomatic, isolated hypercholesterolaemia and if statin treatment could influence this cytokine. METHODS: The investigation was designed as a cross sectional study followed by a single, blind, treatment study of patients receiving pravastatin 80 mg/day for 8 weeks. The study included 23 patients with severe hypercholesterolaemia (LDL>5.2 mmol/L) and 39 normocholesterolaemic controls. Blood samples were obtained from patients and controls at baseline and from patients at end of the study and analysed for lipoproteins and inflammatory mediators: MCP-1. high-sensitivity C-reactive protein (HS-CRP). Isolated peripheral mononuclear cells were analysed for CCR-2 gene expression. RESULTS: Mean plasma LDL-C was significantly higher in patients than in controls. No difference in plasma MCP-1 levels or CCR-2 gene expression was seen between the groups at baseline, nor were there any differences in plasma concentrations of CRP. After treatment with pravastatin, LDL-C decreased by 31%. Treatment did not significantly affect the levels of MCP-1 or CCR-2 gene expression, nor was CRP affected by treatment with pravastatin. CONCLUSIONS: Our study does not support the view that MCP-1 plasma levels and CCR-2 gene expression in circulating monocytes are directly responsible for the monocyte recruitment into the arterial intima in patients with severe asymptomatic hypercholesterolaemia. In addition, the inflammatory response of a high concentration of LDL-C in isolated asymptomatic hypercholesterolaemia is minute. PMID- 14743962 TI - Evaluation, including effects of storage and repeated freezing and thawing, of a method for measurement of urinary creatinine. AB - The aims of this study were to elucidate to what extent storage and repeated freezing and thawing influenced the concentration of creatinine in urine samples and to evaluate the method for determination of creatinine in urine. The creatinine method was based on the well-known Jaffe's reaction and measured on a COBAS Mira autoanalyser from Roche. The main findings were that samples for analysis of creatinine should be kept at a temperature of -20 degrees C or lower and frozen and thawed only once. The limit of detection, determined as 3 x SD of 20 determinations of a sample at a low concentration (6.1 mmol/L), was 0.3 mmol/L, and the recovery of a certified reference material was 97%. The relative precision at 3.15 mmol/L was 2.3%. It was concluded that the method is appropriate for measurement of urinary creatinine. PMID- 14743963 TI - On the agreement between two diagnostic methods with binary outcomes. PMID- 14743964 TI - Commentary on the dual-vision method for analysis of agreement data. PMID- 14743965 TI - In vivo interaction between ciprofloxacin hydrochloride and the pulp of unripe plantain (Musa paradisiaca). AB - The absorption of quinolone antibiotics is seriously impaired by polyvalent cations due to the formation of unabsorbable complexes. M. paradisiaca Linn. (Musaceae), an important staple food in most parts of the world, has been demonstrated to contain many polyvalent cations. The aim of the work was to study the influence of concurrent administration of M. paradisiaca and ciprofloxacin HCI on the pharmacokinetic profiles of ciprofloxacin. The study was carried out in two phases in five healthy male rabbits. Phase one involved oral administration of ciprofloxacin (40 mg/kg) to rabbits, withdrawing blood from the marginal ear vein at 1, 2, 4, and 24 h intervals and checking the serum ciprofloxacin concentration. After a one-week drug "wash-out" period, the second phase started with concurrent oral administration of M. paradisiaca (800 mg/kg) and ciprofloxacin (40 mg/kg). Blood was again withdrawn and analyzed for serum ciprofloxacin content. Antimicrobial activity of the serum was also assessed and expressed as reciprocal serum inhibitory titer. Co-administration of both agents resulted in significant (P<0.05) decrease in serum concentration of ciprofloxacin at all the time interval except at the 24th hour. The following pharmacokinetic parameters were also decreased: area under the curve (81.53%), peak serum concentration (94.37%), elimination rate constant (42.35%); while increase in half-life (81.08%) and clearance rate (69.64%) were noted. Antimicrobial study showed that the antimicrobial potency against E. coli was also decreased by such concurrent administration. The pharmacokinetic parameters and antimicrobial activities of ciprofloxacin were significantly decreased when it was given concurrently with pulp of unripe plantain. Complex formation between the drug and the polyvalent cations present in plantain, leading to decrease in absorption and hence bioavailability, may be responsible for the observed antagonistic interactions. PMID- 14743966 TI - Pharmacokinetics of magnesium glycyrrhizinate following intravenous administration of magnesium glycyrrhizinate in rats. AB - The elimination, distribution, excretion of magnesium glycyrrhizinate (MG) after intravenous (i.v.) administration in rats, and the binding rate of MG to plasma protein were investigated. The concentrations of MG in plasma, tissue, and excretion of rats after i.v. administration of MG were measured using reversed phase high performance liquid chromatography (RP-HPLC). The concentrations of MG in plasma declined in an apparent biexponential manner. The pharmacokinetic parameters from a two-compartment model analysis of plasma samples after i.v. administration of MG 30, 60, and 120 mg/kg-, were t(1/2beta) (min): 140, 180, and 240; AUC(0 approximately 18) (g x min x L(-1)): 10212, 15432, and 50321; CL (L x kg(-1) x min(-1)): 0.0025, 0.0039, and 0.0021, respectively. The drug administered as an iv injection, were mainly cumulated in the liver. When MG was administered to bile-duct cannulated rats, about 90% of i.v. dosed MG was excreted into bile under unchanged form within 24 h after administration. The average binding rate of MG to plasma protein was 87%. The experimental results showed that the distributional property of MG in the present rats study is beneficial to its liver protective activity and liver function improvement. PMID- 14743967 TI - Identification of phase I and phase II metabolites of Guanfu base A hydrochloride in human urine. AB - Guanfu base A is a novel arrhythmic drug candidate isolated from the tuber of a traditional Chinese herb. Phase I and Phase II metabolites of Guanfu base A (GFA) Hydrochloride were studied in human urine by means of liquid chromatography mass spectrometry (LC/MSD) and tandem mass spectrometry (MS/MS). For phase I metabolites, Guanfu base I (GFI) was separated by HPLC and identified by comparison with authentic reference for their retention times, molecular ion peaks, fragment ions, and UV spectra. GFA oxide was also indicated to exist in human urine. For phase II metabolites, after human urine was treated either with glucuronidase or sulfatase, GFA occured in the chromatograms. It was suggested that there were GFA glucuronide and GFA sulfate in human urine. Further more, positive molecular ions, m/z 606 and m/z 510, of the two conjugates were detected in human urine by LC/MSD. In addition, characteristic ion of m/z 606 was identified as the precursor ion of m/z 177 [Glucuronic acid+H]+ by using MS/MS. Characteristic ion of m/z 430 [GFA+H]+ was also identified as a product ion of m/z 606 [GFA glucuronide+H]+. It was concluded that there were GFI. GFA oxide, GFA glucuronide and GFA sulfate in human urine. PMID- 14743968 TI - In vivo metabolism of N-phenyl-N'-(3,5-dimethylpyrazole-4-yl) thiourea in rats. AB - Thiourea derivatives have been shown to posses several biological activities. Therefore a series of N-substituted -N'-(3,5-di/1,3,5trimethylpyrazole-4 yl)thioureas were synthesized and the antitubercular and anticonvulsant activities were studied. Among the new compounds, N-phenyl-N'-(3,5 dimethylpyrazole4-yl)thiourea (S) was demonstrated to have remarkable anticonvulsant activity. In this study, S was selected as a model compound and the in vivo metabolic pathway in rats was investigated. The substrate was given intraperitoneally (50 mg/kg) and blood samples were collected at 0, 1, 2, 4, 8, 12, 24 and 48 h. The substrate and its potential metabolites were separated using HPLC on reverse phase system. The substrate was detected at all times with small quantity of metabolites. PMID- 14743969 TI - Hepatic metabolism of the new antipsychotic agent, mazapertine, in rat--API-MS/MS identification of metabolites. AB - The In vitro biotransformation of the antipsychotic agent, mazapertine was studied after incubation with rat hepatic S9 fraction in the presence of an NADPH generating system. Unchanged mazapertine (42% of the sample) plus 12 metabolites were profiled, quantified, and tentatively identified on the basis of API (ionspray)-MS/MS data. The proposed metabolic pathways for mazapertine are proposed, and the 6 metabolic pathways are: (1) phenylhydroxylation, (2) piperidyl oxidation, (3) O-dealkylation, (4) N-dephenylation, (5) oxidative N debenzylation, and (6) dehydration. Pathways 1 to 3 formed 4-OH-phenyl (M1, 10%) and 4-OH-piperidyl (M2, 9%)-mazapertine, O-desisopropyl mazapertine (M3, 17%), and N-desbenzoylpiperidine-mazapertine (M8, 14%) as 4 major metabolites. Mazapertine is extensively metabolized in rat hepatic S9 fraction. PMID- 14743970 TI - Pharmacokinetic evaluation of guar gum-based colon-targeted oral drug delivery systems of metronidazole in healthy volunteers. AB - The present study was carried out to find the in vivo performance of guar gum based colon-targeted tablets of metronidazole as compared to an immediate release tablets in human volunteers. Six healthy volunteers participated in the study and a crossover design was used. Blood samples were obtained at different time intervals and the plasma concentration of metronidazole was estimated by reverse phase HPLC. The immediate release tablets of metronidazole produced peak plasma concentration (Cmax of 2990 +/- 574.6 ng/mL) within 2.8 +/- 0.6 h. On oral administration of colon-targeted tablets, metronidazole started appearing in the plasma between 5 h and 8 h, and reached the peak concentration (Cmax of 1940.0 +/ 528.4 ng/mL) at 11.1 +/- 2.1 h (Tmax). The AUC(0-infinity) and t(1/2) of metronidazole were unaltered on administering the drug as a colon-targeted tablet indicating that the extent of absorption and elimination were not affected by targeting the drug to the colon. However, colon-targeted tablets showed delayed tmax and absorption time (ta), decreased Cmax and decreased absorption rate constant as compared to immediate release tablets. This in turn indicated that metronidazole was delivered to the colon resulting in a slow absorption of the drug and making it available for local action in the colon. PMID- 14743971 TI - Interspecies variability and drug interactions of loxapine metabolism in liver microsomes. AB - Loxapine is a dibenzoxazepine neuroleptic that is metabolized by the liver in humans. In the present study, we investigated first in vitro loxapine metabolism in liver microsomes from various species including rats, mice, guinea pigs, dogs, rabbits, monkeys and humans. This enables us to choose between species to further validate drug-drug interaction studies. We observed the formation of desmethyl- and hydroxy- metabolites of loxapine after incubation of the different species liver microsomes. Hydroxylation pathway was major in all species. Wide interspecies variability of loxapine metabolism was observed. Loxapine metabolism was similar in human, guinea pig and dog microsomes. We screened in vitro effects of 67 molecules, representative of 8 therapeutic classes, on loxapine metabolism. Loxapine (100 microM) was incubated with guinea pig liver microsomes (1 mg/ml) 30 min at 37 degrees C with and without the presence of interacting drug. We found that most of psychotropics (alimemazine, cyamemazine and levomepromazine), antifungal (ketoconazole), anticancer drugs (daunorubicin, pirarubicin) and analgesic (nefopam) inhibited more than 50% of hydroxyloxapine formation in vitro. Complementary clinical and pharmacokinetic studies should be performed to confirm these results. PMID- 14743972 TI - Improving of the accuracy of in vitro-in vivo linear correlation using kinetic models for ultra sustained release theophylline tablets. AB - The objective of the current study was to establish and evaluate a new technique to increase the accuracy of the in vitro/in vivo linear correlation of single and multiple dose ultra-sustained release theophylline (USRT) preparation (Xantium) in hospitalized patients. In vitro dissolution data for theophylline were collected for 24 h using a USP I (basket) and USP II (paddle) methods. In vivo plasma concentration data were obtained from 8 patients after administration of either single or multiple doses of theophylline. Both in vitro and in vivo results were evaluated by zero-order, first-order, RRSBW, Hixson-Crowell, Higuchi, Hopfenberg, Langenbucher, modified Langenbucher and (Bt)a kinetic models. The individual linear correlations between each in vitro and in vivo percent results and their kinetic distributions were established and regression equations were obtained. The determination coefficient results of the linear kinetic correlations were found to be 0.994 and 0.997 for single and multiple doses by basket method and 0.992 and 0.998 for single and multiple doses by paddle method, respectively. Furthermore, the results of the linear correlations were found as 0.953 and 0.950 for single and multiple doses by basket method and 0.963 and 0.962 for single and multiple doses by paddle method respectively. Therefore, this study suggested that the accuracy of the linear correlation could be improved signilicantly by using linear kinetic correlation. PMID- 14743973 TI - Effect of P-glycoprotein inhibitors erythromycin and cyclosporin A on brain pharmacokinetics of nimodipine in rats. AB - Effect of P-glycoprotein (P-gp) inhibitors erythromycin (Ery) and cyclosporin A(CsA) on brain pharmacokinetics of nimodipine (NMD) in rats was studied. NMD concentrations in rat plasma and brain were determined after iv 2 mg/kg NMD alone, co-administration with Ery and CsA, respectively. It was found that concentrations of NMD in plasma of the three groups had a little difference, but brain concentrations of NMD in rats co-administrated with Ery and CsA were significantly higher than those in rats NMD alone. Significances of NMD in rat brain were found at 20, 40, 60 and 90 min after iv NMD. The brain T(1/2) in rat treated with ery(75.0 min) and CsA(79.0 min) were larger than that (44.2 min) in rats NMD alone. The results indicated that P-gp inhibitors Ery and CsA may increase concentration in rat brain by inhibiting elimination of NMD from brain. PMID- 14743974 TI - Mechanism of the stimulatory effect of 6-aminohexanoic acid on plasminogen activation by streptokinase or tissue plasminogen activator: the role of chloride. AB - Studies were conducted on the mechanism of the stimulatory effect of 6 aminohexanoic acid (6-AH) during the in vitro activation of human glutamic plasminogen (Glu-Plg) by streptokinase or by tissue plasminogen activator (t-PA) and the possible role of the addition of physiological concentrations of NaCl to the buffer solution. Enhancement by 6-AH was investigated by measuring the rate of plasmin generation using chromogenic substrate H-D-glu-phe-lys-pNA (S-2403). Control studies using plasmin showed that the addition of 6-AH at concentrations below 20 mM did not significantly affect the initial rate of the amidolytic activity of plasmin with or without the addition of NaCl to 0.05 M Tris buffer (pH 7.4). On the other hand, addition of NaCl to the buffer slowed down the initial rate of activation of Glu-Plg by streptokinase or by t-PA while increasing the percent enhancement by 6-AH when compared with the controls. The ratios of the initial rates of plasmin generation in the presence or in the absence of 6-AH were plotted against the inverse of the volume fraction of Glu Plg, streptokinase or t-PA after serial dilutions. The results showed that when the activation reactions were performed in 50 mM of Tris buffer (pH 7.4), the enhancements by 6-AH were related to its interaction with streptokinase or t-PA, while using the same Tris buffer containing 0.6 % NaCl, the enhancements by 6-AH were related to its interaction with both Glu-Plg and streptokinase or t-PA. However, upon increasing the NaCl to 0.9%, the results showed that the enhancements by 6-AH of the activation of Glu-Plg by streptokinase or t-PA were related to its interaction with Glu-Plg. The results suggested that changes in the concentrations of NaCl play a regulatory role during the activation process. PMID- 14743975 TI - The properties and functions of bacterial aminopeptidases. AB - Aminopeptidases are enzymes that release N-terminal amino residues from oligopeptides, polypeptides and proteins. The classification of aminopeptidases has often been based on mechanism of catalysis, structure of active site, substrate specificity kinetic and molecular properties. In terms of catalytic mechanism bacterial aminopeptidases can be divided into three main catalytic groups: metallo-, cysteine- and serine aminopeptidases. According to their substrate specificity the enzymes can be ordered into two sub-groups: having broad or narrow specificity. Almost half of the characterized aminopeptidases show a subunit structure. Enzymes having a quaternary structure are most often built of a combination of 2, 4, 6 subunits. Bacterial aminopeptidases may be localised in the cytoplasm, on membranes, associated with the envelope or secreted into the extracellular media. Regulation of the synthesis of aminopeptidases is assumed to take place mainly at the level of transcription. Most genes encoding the enzymes are monocistronic and contain a promotor characteristic for the genes transcribed by RNA polymerase associated with the factor sigma70. Aminopeptidases play an important role in the initial and final steps of protein turnover and they are involved in several specific regulatory functions. PMID- 14743976 TI - Horizontal DNA transfer between bacteria in the environment. AB - In the environment horizontal DNA transfer between various bacterial species and genera takes place by transformation, transduction, but mainly by conjugation. Conjugation is responsible for the spread of genes coding for antibiotic resistance and xenobiotic degradation. Transfer events are reported in animal, rhizosphere and phylloplane ecosystems and in non polluted and polluted water and soil. Genetic exchange between Bacteria and Archaea is also observed. Evaluation of the extent of interspecies gene transfer is crucial in view of the deliberate release of a variety of unmodified and genetically modified microorganisms into the natural environments. PMID- 14743977 TI - Rearrangements between differently replicating DNA strands in asymmetric bacterial genomes. AB - Many bacterial genomes are under asymmetric mutational pressure which introduces compositional asymmetry into DNA molecule resulting in many biases in coding structure of chromosomes. One of the processes affected by the asymmetry is translocation changing the position of the coding sequence on chromosome in respect to the orientation on the leading and lagging DNA strand. When analysing sets of paralogs in 50 genomes, we found that the number of observed genes which switched their positions on DNA strand is lowest for genomes with the highest DNA asymmetry. However, the number of orthologs which changed DNA strand increases with the phylogenetic distance between the compared genomes. Nevertheless, there is a fraction of coding sequences that stay on the leading strand in all analysed genomes, whereas there are no sequences that stay always on the lagging strand. Since sequences diverge very fast after switching the DNA strand, this bias in mobility of sequences is responsible, in part, for higher divergence rates among some of coding sequences located on the lagging DNA strand. PMID- 14743978 TI - Synthesis of siderophores by strains of Staphylococcus cohnii isolated from various environments. AB - Siderophore activity as the feature of microorganisms enabling colonization of human body and the survival in inanimate environment was investigated in 108 strains of Staphylococcus cohnii; S. cohnii ssp. cohnii (50 strains) and S. cohnii ssp. urealyticus (58 strains). Strains were isolated from people, hospital and non-hospital environment. Highest siderophore activity was noted in strains S. cohnii ssp. urealyticus particularly from the inanimate environments origin. In 86% analyzed strains siderophores of hydroxamate class were detected. Larger amounts of these compounds were synthesized in strains S. cohnii ssp. urealyticus. Strains belonging to both subspecies from human origin showed lower activity of siderophores (total pool) and did not produce hydroxamate class chelators or produced very small amounts of these compounds. PMID- 14743979 TI - Differences in inhibition of apoptosis depending on the virulence of used herpes simplex virus type 1 strains. Function of interferon alpha in apoptotic death of virus infected cells. AB - The purpose of the present study was to determine whether there is the relation between the virulence of used HSV-1 strains and inhibition of apoptosis. HEp-2 cells were induced to apoptosis by osmotic shock after infection by HSV-1 strains. HSV-1 ts, earlier described as less virulent for mice inhibited apoptosis in smaller degree than native strain and HSV-1 tr. We suggest that this is due to the hyperproduction of IFN alpha by human cells after the stimulation by this strain. All strains of HSV-1 didn't inhibit apoptosis in the presence of IFN alpha and apoptosis was inhibited by anti IFN alpha antibodies. We confirm that IFN alpha plays an important function in controlling acute HSV-1 infection. PMID- 14743980 TI - Enteropathogenic activity and invasion of HEp-2 cells by Aeromonas caviae clinical isolates. AB - Twenty Aeromons caviae isolates from stool of children with diarrhea symptoms were examined for virulence-associated properties: production of cytotoxic and cytotonic toxins, and invasive ability. Most of A. caviae strains were cytotoxic to Vero and CHO cells and produced cytotonic toxins which caused elongation of CHO cells. Moreover, five of A. caviae strains revealed invasive ability towards HEp-2 cells. PMID- 14743981 TI - Amlodipine: a cardiovascular drug with powerful antimicrobial property. AB - Ten cardiovascular drugs were procured in pure form from their manufacturers in India and screened for antimicrobial property against fifteen known bacteria belonging to both gram-positive and gram-negative types. These bacteria were inhibited by the common antibiotics at 1-5 mg ml(-1) level through our earlier studies. Since most of the bacteria were moderate to highly responsive to amlodipine, this compound was further tested in vitro against 504 bacteria comprising 4 genera of gram-positive and 15 genera of gram-negative bacteria. Most of these were inhibited by the drug at 50-200 microg ml(-1) level and few strains were sensitive even at lower concentrations (10 microg ml(-1)). The bacteria could be arranged in the decreasing order of sensitivity towards amlodipine in the following manner: Staphylococcus aureus, Vibrio cholerae, Vibrio parahemolyticus, Shigella spp., Salmonella spp., Bacillus spp., whereas Escherichia coli, Klebsiella spp. and Pseudomonas aeruginosa were found to be resistant to the lower concentrations of the drug. Amlodipine was found to be bactericidal in nature when its mode of action was studied against S. aureus 6571, V. cholerae 14035 and Sh boydii 8 NCTC 254/66. The antibacterial activity of amlodipine could also be confirmed in vivo. When it was given to Swiss strain of white mice at different dosages (30 and 60 microg/mouse), it could significantly protect the animals challenged with 50 MLD of Salmonella typhimurium NCTC 74. According to Chi square test the in vivo data were highly significant (p<0.001). PMID- 14743982 TI - Comparative delta-endotoxins of Bacillus thuringiensis against mosquito vectors (Aedes aegypti and Culex pipiens). AB - Pure crystals of seven Bacillus thuringiensis field isolates from the Lower Silesia region (Poland) were tested against larvae of Aedes aegypti L. and Culex pipiens L. (Culicidae, Diptera). The crystals of OpQ3 phylloplane isolate (belonging to the first biochemical type of B. thuringiensis subsp. japonensis, yoso, jinghongiensis) killed from 68 +/- 7% to 84 +/- 7% of the fourth instar larvae of A. aegypti. The crystals of two other strains (KpF3 and KpC1) of this group caused mortality between 3 +/- 2% and 70 +/- 7%. The LC50 ranged from 3.2 +/- 0.4 to 34.1 +/- 4.8 microg/ml. The effect of B. thulringiensis wratislaviensis H-47 crystals was the lowest with larval mortality from 0% to 17 +/- 3%. No significant (0%-37 +/- 6%) effect of B. thuringiensis crystals on the larvae of C. pipiens was observed. Our results show that the delta-endotoxins of B. thuringiensis act very specifically. PMID- 14743983 TI - Improvement of rifemycins production by Amycolatopsis mediterranei in batch and fed-batch cultures. AB - The production of rifamycins B and SV using glucose as main C-source by Amycolatopsis mediterranei in batch and fed-batch culture was investigated. Fed batch culture using glucose as mono feeding substrate either in the form of pulse addition, in case of shake flask, or with constant feeding rate, in bioreactor level, proved to be an alternative production system with a significant increase in both volumetric and specific antibiotic production. The maximal concentrations of about 1146 mg/l and 2500 mg/l of rifamycins B and SV, respectively, was obtained in fed-batch culture in bioreactor level under non-oxygen limitation. On the other hand, the rate of rifamycins production was increased from 6.58 to 12.13 mg/l x h for rifamycin B and from 9.47 to 31.83 mg/l x h for rifamycin SV on the bioprocess transfer and improvement from the conventional batch cultivation in shake flask to fed-batch cultivation in stirred tank bioreactor. PMID- 14743984 TI - The immediate effects of soft tissue mobilization with proprioceptive neuromuscular facilitation on glenohumeral external rotation and overhead reach. AB - STUDY DESIGN: Randomized controlled 2-group. pretest-posttest, multivariate study of patients with shoulder musculoskeletal disorders. OBJECTIVES: The purpose of this study was to evaluate the immediate effect of soft tissue mobilization (STM) with proprioceptive neuromuscular facilitation (PNF) to increase glenohumeral external rotation at 45 degrees of shoulder abduction and overhead reach. BACKGROUND: It is postulated that limitation in glenohumeral external rotation, when measured at 45 degrees of shoulder abduction, represents subscapularis muscle flexibility deficits and is associated with the inability to fully reach overhead. No research, however, is available to demonstrate whether intervention strategies intended to improve subscapularis flexibility and glenohumeral external rotation range of motion at 45 degrees of shoulder abduction will improve a patient's ability to reach overhead. METHODS AND MEASURES: Twenty patients (10 males, 10 females; age range, 21-83 years) with limited glenohumeral external rotation and overhead reach of 1 year duration or less served as subjects. The subjects were randomly assigned to a treatment group, which consisted of soft tissue mobilization to the subscapularis and proprioceptive neuromuscular facilitation to the shoulder rotators, or a control group. Goniometric measurements of glenohumeral external rotation at 45 degrees abduction and overhead reach were taken preintervention and immediately postintervention for the treatment group or at prerest and postrest periods for the control group. RESULTS: The treatment group improved by a mean of 16.4 degrees (95% confidence interval [CI, 12.5 degrees-20.3 degrees) of glenohumeral external rotation, as compared to less than a 1 degree gain (95% CI, -0.2 degrees 2.0 degrees) in the control group (P < .0005). Overhead reach in the treatment group improved by a mean of 9.6 cm (95% CI, 5.2-14.0 cm) in comparison to a mean gain of 2.4 cm (95% CI, -0.8-5.6 cm) for the control group (P = .009). CONCLUSION: These findings suggest that a single intervention session of STM and PNF was effective for producing immediate improvements in glenohumeral external rotation and overhead reach in patients with shoulder disorders. PMID- 14743985 TI - Strength response in human femoris muscle during 2 neuromuscular electrical stimulation programs. AB - STUDY DESIGN: Experimental mixed repeated-measures design. OBJECTIVE: To determine the effect of 2 versus 3 neuromuscular electrical stimulation (NMES) training sessions per week on the response to strength training of the quadriceps femoris muscle. BACKGROUND: Many studies have examined the influence of training intensity (percent maximal voluntary isometric contraction [MVIC]) during NMES on the strength response of the quadriceps femoris muscle. However, no study has examined the effects of the number of NMES sessions per week on the change in strength of the quadriceps femoris. METHODS AND MEASURES: Twenty-seven healthy subjects (mean age +/- SD, 23.2 +/- 3.2 years) volunteered for the study and were randomly assigned to 1 of 3 groups; control group (no electrical stimulation); group 2 (NMES 2 times per week); and group 3 (NMES 3 times per week). Groups 2 and 3 received NMES (10 minutes per session) over a 4-week period for a total of 8 and 12 NMES training sessions, respectively. The isometric quadriceps femoris muscle force produced during NMES was monitored during each treatment minute. The MVIC force of the quadriceps femoris was assessed prior to the first week and at the start of weeks 2, 3, and 4 of the 4-week training program, with a final measurement after the fourth week (5 total measurements) for all subjects. RESULTS: Only the mean percent change in quadriceps MVIC before and after the 4 weeks of training with NMES between the control group and group 3 was significantly different (P = .021). CONCLUSIONS: Based on the electrical stimulation parameters and healthy subjects used in this study, NMES caused significant increases in the quadriceps femoris muscle strength when used for 3 training sessions per week for 4 weeks. It is possible that the use of a different electrical stimulation paradigm and/or a different patient population may result in strength gains with 1 or 2 sessions per week. PMID- 14743986 TI - The effect of static stretch and warm-up exercise on hamstring length over the course of 24 hours. AB - STUDY DESIGN: Experimental pretest-posttest control design. OBJECTIVES: The purpose of the study was twofold: (1) to determine the lasting effect of static stretch on hamstring length for up to 24 hours and (2) to compare the efficacy of static stretch with and without warm-up exercise on hamstring length. BACKGROUND: Research is limited on the lasting effects of static stretching and is controversial on the combined effects of warm-up activities and static stretching on muscle lengthening. METHODS AND MEASURES: Fifty-six volunteer subjects (ages 18-42 years) with limited bilateral hamstring length were assigned to 1 of 4 groups: (1) warm-up and static stretch, (2) static stretch only, (3) warm-up only, and (4) control. The warm-up was 10 minutes of stair climbing at 70% of maximum heart rate. Static stretch consisted of a single session of three 30 second passive stretches of the hamstring. Hamstring length was measured preintervention and at several intervals postintervention (immediately and then at 15 minutes, 60 minutes, 4 hours, and 24 hours) using the active knee extension (AKE) test. Data were analyzed using a mixed-model analysis of variance. RESULTS: The warm-up-and-static-stretch group and the static-stretch-only group showed a significant increase in hamstring length between preintervention and all postintervention measurements. At 24 hours poststretch, the warm-up-and-static stretch group had a mean increase of 10.3 degrees (95% confidence interval, 7.7 12.9) and the static-stretch-only group had a mean increase of 7.7 degrees (95% confidence interval, 4.7-10.7) in AKE range of motion (ROM). Both of these groups did show significant decrease (2.9 degrees and 4.0 degrees, respectively) in hamstring muscle length (AKE ROM) at 15 minutes poststretch when compared to immediate poststretch values. The static-stretch-only and the warm-up-and-static stretch groups did not differ significantly from each other. Control and warm-up only groups showed no significant increase in hamstring length between preintervention and any of the postintervention measurements. CONCLUSIONS: A significant increase in hamstring length can be maintained for up to 24 hours when using static stretching. Muscle length gains are greatest immediately after stretching and decline within 15 minutes. The addition of a warm-up exercise prior to stretching does not appear to significantly increase the effectiveness of static hamstring stretching. PMID- 14743987 TI - Test-retest reliability of an abbreviated self-report overall health status measure. AB - STUDY DESIGN: Test-retest reliability study. OBJECTIVE: To assess test-retest reliability and estimate minimal detectable change of an overall measure and 2 summary measures of patient self-report of health status. BACKGROUND: Change in patient self-report of health status is a common outcome measure following rehabilitation. Because collection of health status data takes time and clinicians are required to be productive, selected items from reliable instruments were used to form a new, abbreviated instrument of health status relevant to patients in outpatient rehabilitation. There are no test-retest reliability statistics of these health status measures in this population. METHODS AND MEASURES: A convenience sample of 71 patients (mean age +/- SD, 41.9 +/- 17.9 years; age range, 15-83 years; sex, 35% male), with a variety of orthopaedic diagnoses, seeking rehabilitation in 2 outpatient facilities, volunteered. Patients completed health status questionnaires at initial evaluation and at 24 to 72 hours following evaluation. Intraclass correlation coefficients (ICC2.1) were used to estimate test-retest reliability and to estimate measurement error and minimal detectable changes. RESULTS: ICCs with a 1 sided lower limit 95% confidence intervals (CI) of the Overall Health Status measure and the Physical and Mental Component Summary measures for patients with chronic symptoms were 0.92 (0.85), 0.82 (0.68), and 0.85 (0.74), respectively. Minimal detectable changes (90% CI) were +/-12 (scale range, 100), +/- 9 (scale range, 60), and +/- 9 (scale range, 60) scale points, respectively, for the same measures. CONCLUSIONS: Results support the test-retest reliability of the Overall Health Status measure and summary measures for patients with chronic symptoms and demonstrate ability of the Overall Health Status and Physical Summary Scale measures to detect improvement of patient self-report of health status within the first few days of rehabilitation. PMID- 14743988 TI - Physical activity and its relationship to physical performance in patients with end stage knee osteoarthritis. AB - STUDY DESIGN: Cross-sectional observational design. OBJECTIVES: To compare physical activity levels in men and women with end-stage knee osteoarthritis to those of a comparison group and to examine the relationship between physical activity level and physical performance. BACKGROUND: Osteoarthritis of the knee is associated with significant losses in functional performance and high social costs. Although reductions in physical activity are reported, they have not been quantified or explored. METHODS AND MEASURES: Fifty-nine candidates awaiting total knee arthroplasty (TKAC group) and 79 individuals without osteoarthritis (comparison group) participated. Physical activity was assessed using the Voorrips Questionnaire. Performance measures included fast self-paced walk test, timed up-and-go test, and a timed stair performance measure. A subset of subjects completed the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) and underwent muscular strength and endurance testing. The effects of gender and group were tested using GLM ANOVA. Pearson product moment correlations were used to examine relationships between the variables. RESULTS: All aspects of physical activity were lower (P<.001) in the TKAC group, with a moderate difference in household score (18%) and a large difference in leisure activities (63%). Unlike the comparison group, modest but significant correlations (r = 0.31 0.33, P<.03) were observed between overall physical activity and performance test scores for the TKAC group. Physical activity was not significantly related to pain reported on the WOMAC or during the performance tasks. CONCLUSIONS: The belief that pain limits the physical activity of patients with severe osteoarthritis requires further investigation. The profound differences between a comparison group and patients with end-stage osteoarthritis in physical activity have critical implications for the well-being and effective treatment of this population. PMID- 14743989 TI - Cartilage tissue engineering: controversy in the effect of oxygen. AB - Articular cartilage lacks the ability to repair itself and consequently defects in this tissue do not heal. Tissue engineering approaches, employing a scaffold material and cartilage producing cells (chondrocytes), hold promise for the treatment of such defects. In these strategies the limitation of nutrients, such as oxygen, during in vitro culture are of major concern and will have implications for proper bioreactor design. We recently demonstrated that oxygen gradients are indeed present within tissue engineered cartilaginous constructs. Interestingly, oxygen, besides being an essential nutrient, is also a controlling agent of developmental processes including cartilage formation. However, the specific role of oxygen in these processes is still obscure despite the recent advances in the field. In particular, the outcome of published investigations is inconsistent regarding the effect of oxygen tension on chondrocytes. Therefore, this article describes the possible roles of oxygen gradients during embryonic cartilage development and reviews the data reported on the effect of oxygen tension on in vitro chondrocyte proliferation and differentiation from a tissue engineering perspective. Furthermore, possible causes for the variance in the data are discussed. Finally, recommendations are included that may reduce the variation, resulting in more reliable and comparable data. PMID- 14743990 TI - Homogeneous biocatalysis in organic solvents and water-organic mixtures. AB - Biocatalysis in non-aqueous media has undergone tremendous development during the last decade, and numerous reactions have been introduced and optimized for synthetic applications. In contrast to aqueous enzymology, biotransformations in organic solvents offer unique industrially attractive advantages, such as: drastic changes in the enantioselectivity of the reaction, the reversal of the thermodynamic equilibrium of hydrolysis reactions, suppression of water-dependent side reactions, and resistance to bacterial contamination. Currently, the field is dominated by heterogeneous biocatalysis based primarily on lyophilized enzyme powders, cross-linked crystals, and enzymes immobilized on inert supports that are mainly applied in enantioselective synthesis. However, low reaction rates are an inherent problem of the heterogeneous biocatalysis, while the homogeneous systems have the advantage that the elimination of diffusional barriers of substrates and products between organic and water phases results in an increase in the reaction rate. Here the discussion is focused on the correlation between activity and structure of the intact enzymes dissolved in neat organic solvents, as well as modifications of natural enzymes, which make them soluble and catalytically active in non-aqueous environment. Factors that influence conformation and stability of the enzymes are also discussed. Current developments in non-aqueous biocatalysts that combine advantages of protein modification and immobilization, i.e., HIP plastics, enzyme chips, ionic liquids, are introduced. Finally, engineering enzymes for biotransformations in non conventional media by directed evolution is summarized. PMID- 14743991 TI - Common-path interferometry with one-dimensional periodic filters. AB - We discuss a spatial filtering interferometry setup that employs a periodic spatial filter with either cosine transmittance or binary phase modulation. The setup's input plane is formed by two separate windows, one of which supports a phase object and the other, a reference beam. Using the appropriate frequency and orientation of the filter produces an interference pattern of the two input fields at the output plane of the system. The main attributes and advantages of the setup are discussed and experimentally illustrated with the example of a binary phase periodic filter implemented with a spatial light modulator. PMID- 14743992 TI - Ince-Gaussian beams. AB - We demonstrate the existence of the Ince-Gaussian beams that constitute the third complete family of exact and orthogonal solutions of the paraxial wave equation. Their transverse structure is described by the Ince polynomials and has an inherent elliptical symmetry. Ince-Gaussian beams constitute the exact and continuous transition modes between Laguerre and Hermite-Gaussian beams. The propagating characteristics are discussed as well. PMID- 14743993 TI - Localized subluminal envelope pulses in dispersive media. AB - The existence and properties of propagative nondispersive and nondiffracting localized envelope waves in dispersive transparent media, with either normal or anomalous group-velocity dispersion, showing subluminal group velocities and strong spatial localization are pointed out. Nonparaxial effects may lead to group-velocity reduction of several orders of magnitude. Numerical simulations are given for wave propagation in sapphire. PMID- 14743994 TI - High-contrast white-light Lau fringes. AB - We present a new optical assembly with which to achieve Lau fringes with totally incoherent illumination. Gratinglike codification of the spatially incoherent source combined with an achromatic Fresnel diffraction setup allows us to achieve Lau fringe-pattern visibility of almost 100% with broadband light. The white light character to our proposed setup is in stark contrast to previous monochromatic implementations. Potential implications of this fact are identified. PMID- 14743995 TI - Single-mode Raman fiber laser based on a multimode fiber. AB - Single-mode operation of a Raman fiber laser based on a multimode fiber has been demonstrated for what is to our knowledge the first time. Single-mode operation was achieved through the beam cleanup effect of stimulated Raman scattering in multimode fibers. The Raman laser, consisting of a 50-microm-core multimode fiber and a multimode Nd:YAG laser pump, produced an output beam corresponding to the fundamental mode of the fiber. PMID- 14743996 TI - Trans-4-stilbenemethanol-doped photosensitive polymer fibers and gratings. AB - We report the development of photosensitive polymeric optical fibers for gratings. The fiber is made of a copolymer of methyl methacrylate and butyl methacrylate, and its core is made of a copolymer of methyl methacrylate, ethyl methacrylate, and benzyl methacrylate (90/4/6 molar ratio) doped with trans-4 stilbenemethanol. Ultraviolet absorption and photoinduced refractive-index change of the doped core system are presented, and its photosensitivity is demonstrated by fabrication of Bragg gratings in the core of the fiber by use of a pulse laser at 325 nm. PMID- 14743997 TI - Anomalous intensity of pinned speckles at high adaptive correction. AB - Ground-based optical searches for faint stellar or planetary companions about other stars may be limited by speckle noise, which is the rapid intensity fluctuations that are due to motions of remnant atmospheric speckles. Adaptive optics (AO) can reduce residual wave-front phase errors to low values, substantially reducing the unwanted power in the speckle halo. At high correction, however, the noise in the halo will be dominated by anomalously bright "pinned" speckles that have a number of unusual properties. They can have negative intensities and will appear in spatially antisymmetric patterns; they are spatially pinned to Airy rings and have zero mean in a sufficiently long integration. Some of these properties may be used to reduce the unanticipated effect of pinned speckles on companion searches, depending on details of the AO system. But, in short exposures, pinned speckles dominate speckle noise over much of the inner halo for Strehl ratios S as low as 0.6 and over much of the outer halo too as Strehl and deformable-mirror actuator densities increase. I show that these anomalously bright pinned speckles are not included in the traditional expression for speckle power in an image, (1 - S), on which sensitivity estimates of future high-performance AO systems have been based. PMID- 14743998 TI - Distributed feedback-distributed Bragg reflector coupled cavity laser with a Ti:(Fe:)Er:LiNbO3 waveguide. AB - A thermally fixed photorefractive Bragg grating is written in a single-mode Ti:Fe:Er:LiNbO3 channel waveguide and used to develop a distributed feedback distributed Bragg reflector coupled cavity laser with a second broadband dielectric cavity mirror. The optically pumped (lambda(p) = 1480 nm, P = 130 mW) laser emits in single-frequency operation as much as 8 mW at lambda = 1557.2 nm with a slope efficiency of approximately 22%. The laser wavelength can be thermo optically and electro-optically tuned over 100 pm. PMID- 14743999 TI - Continuous-wave and pulsed laser performance of Nd:LuVO4 crystal. AB - Received August 11, 2003 We report continuous-wave and actively Q-switched laser performance achieved with Nd:LuVO4 crystal for the 4F(3/2) --> 4I(1/2) transition (corresponding wavelength of 1065.8 nm) under high-power diode pumping. Continuous-wave output power of 12.55 W is obtained with an optical conversion efficiency of 50.2%. In actively Q-switched operation the average output power reaches 5.42 W at a pulse repetition frequency of 40 kHz with 18 W of pump power incident upon the crystal, yielding an optical conversion efficiency of 30.1%. The pulse energy and peak power reach 138 microJ and 16.2 kW, respectively, at a pulse repetition frequency of 25 kHz under a pump power of 14.2 W; the pulse duration is 8.5 ns. PMID- 14744000 TI - Real-time measurement of in vitro flow by Fourier-domain color Doppler optical coherence tomography. AB - The possibility of measuring a full Doppler flow depth profile in parallel by use of frequency-domain optical coherence tomography is demonstrated. The method is based on a local phase analysis of the backscattered signal and allows for imaging of bidirectional Doppler flow. The Doppler frequency limit is 5 kHz for the presented measurements and is set by half of the frame rate of the CCD detector array. We measured the flow of 0.3-microm microspheres suspended in distilled water at controlled flow rates and in vitro human blood flow through a 200-microm capillary with a real-time color-encoded Doppler tomogram rate of 2 3/s. PMID- 14744001 TI - Near-field visualization of light confinement in a photonic crystal microresonator. AB - By using scanning near-field optical microscopy, we directly map the subwavelength confinement of light around a point defect in a two-dimensional photonic crystal microresonator. Comparison of our results with the outcome of three-dimensional finite-difference time domain calculations allows us to identify small imperfections in the structure that result in the spatial modification of the intensity distribution. PMID- 14744002 TI - Piezo-driven deformable mirror for femtosecond pulse shaping. AB - We have developed a deformable, gold-coated mirror based on piezoelectric actuators with 15-micros response time. With 20 independent channels we were able to compress 72-fs pulses from a Ti:sapphire oscillator down to 45 fs in a 4f zero dispersion compressor arrangement. Spectral interference was used to measure the mirror performance, while the spectral phase interferometry for direct electric field reconstruction (SPIDER) technique was used for the laser pulse characterization. PMID- 14744003 TI - Properties of two interlinked chi(2) interactions in noncollinear phase matching. AB - We present a compact experimental realization of the interaction among five field modes in a chi(2) nonlinear crystal. The classical evolution of the fields can be analytically described assuming that two of the fields play the role of nondepleted pumps. If one of the fields has a nonzero input amplitude, then the other two fields at the output are holographic replicas of the input signal. PMID- 14744004 TI - Generalized phase-shifting interferometry with arbitrary unknown phase steps for diffraction objects. AB - A general method of extracting the arbitrary unknown and unequal phase steps in phase-shift interferometry from interferograms recorded on the diffraction field of an object and then reconstructing the object wave front digitally with our derived formulas is proposed. The phase steps are first calculated based on the statistical nature of the diffraction field and are further improved by an iterative approach. This method is simple, highly accurate, and usable for any frame number N (N > or = 3) and for both smooth and diffusing objects, as is verified by a series of computer simulations. PMID- 14744005 TI - Optimal switching from recording to fixing for high diffraction from a LiNbO3:Ce:Cu photorefractive nonvolatile hologram. AB - An oscillatory characteristic of diffraction is observed as a result of strong beam coupling in a weakly oxidized LiNbO3:Ce:Cu crystal during the recording step in the production of nonvolatile photorefractive holograms. On this basis the optimal switching time from the recording step to the fixing step for high diffraction of a fixed hologram is studied. It is shown that switching after the first diffraction maximum leads theoretically to fixed diffraction of as much as 100% with a suitable switching time. Both an experimental demonstration and a theoretical simulation are presented. PMID- 14744006 TI - Networks of local minima in optical system optimization. AB - We discuss a surprising new feature of the merit function landscape in optical system design. When certain conditions are satisfied, the local minima form a network in which all nodes are connected. Each link between two neighboring minima contains a saddle point with a Morse index of 1. For a simple global optimization search (the symmetric Cooke triplet), the network of the corresponding set of local minima is presented. PMID- 14744007 TI - Narrow frequency and sharp angular defect mode in one-dimensional photonic crystals from a photonic heterostructure. AB - By combining two one-dimensional defective photonic crystals (PCs), we obtained a photonic heterostructure with narrow frequency and a sharp angular defect mode. The key to obtaining such a structure is to design the two sub-PCs to make the frequencies of their defect modes the same at one incident angle and different at all other incident angles. Filters designed on the basis of this heterostructure possess not only a narrow-frequency passband but also a sharp angular pass breadth. Optimization of the practical design is also suggested. PMID- 14744008 TI - Planar silicon-based light polarizers. AB - Silicon-based thin-film polarizers operating in the visible and near-infraed spectral range are fabricated by electrochemical etching of bulk silicon wafers. Anisotropically etched (110) porous silicon layers exhibit a strong in-plane anisotropy of the refractive index. Stackes of alternating layers with different mean refractive indices and thicknesses act as dichroic Bragg reflectors or microcavities, respectively. Both structures have two distinct reflection and transmission bands depending on the polarization of the incident linearly polarized light. Planar polarizers are realized through the combination, in one structure, of a dichroic reflector with either a second reflector or a microcavity with different spectral responses. PMID- 14744009 TI - Regenerative amplification of femtosecond laser pulses in Ti:sapphire at multikilohertz repetition rates. AB - We have generated and applied noncoherent x-ray radiation in an all-solid-state laser system operating at repetition rates up to 20 kHz. Based on a model that takes into account the strong thermal loading of the Ti:sapphire rod, a laser cavity with low sensitivity to thermal lensing was chosen. With a maximum pump power of 80 W, an output power as high as 27 W was obtained in gain-switched operation, and, with a seeding from a femtosecond oscillator, 60-fs, 0.8-mJ (8-W) pulses at 10 kHz and 0.32-mJ (6.5-W) pulses at 20 kHz were generated. High power femtosecond output was used to generate x-ray continuum radiation up to 5 keV from a liquid-gallium jet target. PMID- 14744010 TI - Generation of 14.8-fs pulses in a spatially dispersed amplifier. AB - We demonstrate amplification and compression of 110-nm broad spectra in a spatially dispersed amplifier for what is believed to be the first time and generate 14.8-fs pulses with 450 microJ of energy at a repetition rate of 1 kHz. The amplifier concept is scalable in energy and allows for spectral shaping, which was demonstrated and compared with numerical simulations and showed excellent agreement. PMID- 14744011 TI - Chromatic dispersion characterization by direct instantaneous frequency measurement. AB - A new concept for the measurement of chromatic dispersion is presented. The group delay of the device under test is directly obtained from the instantaneous frequency of a short optical pulse after propagation in the device, itself obtained with temporal and spectral interferometry. Temporal and spectral fringe encoding provide excellent accuracy and resistance to noise, and a wide variety of dispersions can be characterized even at high insertion losses. PMID- 14744012 TI - Optimization of high-order harmonic generation by adaptive control of a sub-10-fs pulse wave front. AB - We present a method for the optimization of high-order harmonic generation based on wave-front correction of the driving laser beam. The technique exploits wave front adaptive control by means of a deformable mirror, governed by an optimization procedure. PMID- 14744013 TI - Zero-additional-phase SPIDER: full characterization of visible and sub-20-fs ultraviolet pulses. AB - We demonstrate a novel spectral-shearing interferometry setup for characterizing the temporal amplitude and phase of ultrashort optical pulses over an extremely wide wavelength region. By the mixing of two strongly chirped auxiliary pulses with the pulse to be characterized, two spectrally sheared replicas are generated, and their spectral interference is evaluated. We fully characterize 10 fs pulses in the visible region by sum-frequency mixing and 19-fs pulses in the ultraviolet region by difference-frequency mixing. The scheme is self-referencing and highly sensitive. The zero-additional-phase scheme does not alter the unknown pulses and yields the pulse shape at the interaction point of a spectroscopic experiment. PMID- 14744014 TI - Molecular mechanism for activation of superoxide-producing NADPH oxidases. AB - The membrane-integrated protein gp91phox, existing as a heterodimer with p22phox, functions as the catalytic core of the phagocyte NADPH oxidase, which plays a crucial role in host defence. The oxidase, dormant in resting cells, becomes activated to produce superoxide, a precursor of microbicidal oxidants, by interacting with the adaptor proteins p47phox and p67phox as well as the small GTPase Rac. In the past few years, several proteins homologous to gp91phox were discovered as superoxide-producing NAD(P)H oxidases (Nox's) in non-phagocytic cells; however, regulatory mechanisms for the novel oxidases have been largely unknown. Current identification of proteins highly related to p47phox and p67phox, designated Noxol (Nox organizer 1) and Noxal (Nox activator 1), respectively, has shed lights on common and distinct mechanisms underlying activations of Nox family oxidases. PMID- 14744015 TI - Age-related changes in the activity of antioxidant and redox enzymes in rats. AB - Cellular defense system, including glutathione, glutathione-related enzymes, and antioxidant and redox enzymes, may play crucial roles in the aging of aerobic organisms. To understand the physiological roles of these factors in the aging process, their levels were compared in the livers and brains of 5-week- and 9 month-old rats. GST activity was higher in livers and brains of 9-month-old rats than in those of 5-week-old rats, and brain catalase activity was about 2-fold higher. However, it was unchanged in the livers of the 9-month-old rats. gamma Glutamylcysteine synthetase activity was about 2-fold higher in the brains of the older rats but again not in their livers. In contrast glutathione synthetase activity appeared to be lower in the livers of the older rats while GSH content did not change with age in livers and brains. Glutathione peroxidase activity was higher in 9-month-old rat brains, but lower in 9-month-old rat livers, while superoxide dismutase activity was higher in both tissues in the older rats. The activities of two redox enzymes, thiol-transferase and thioredoxin reductase, did not change with age, nor did that of glutathione reductase. These results indicate that levels of different cellular defense systems vary with age in an irregular manner. PMID- 14744016 TI - Ca2+: a stabilizing component of the transglutaminase activity of Galphah (transglutaminase II). AB - Galphah (transglutaminase type II; tissue transglutaminase) is a bifunctional enzyme with transglutaminase (TGase) and guanosine triphosphatase (GTPase) activities. The GTPase function of Galphah is involved in hormonal signaling and cell growth while the TGase function plays an important role in apoptosis and in cross-linking extracellular and intracellular proteins. To analyze the regulation of these dual enzymatic activities we examined their calcium-dependence and thermal stability in enzymes from several cardiac sources (mouse heart, and normal, ischemic and dilated cardiomyopathic human hearts). The GTP binding activity of Galphah was markedly inhibited by Ca2+ whereas the TGase activity was strongly stimulated, suggesting that Ca2+ acts as a regulator, switching Galphah from a GTPase to a TGase. The TGase function of Galphah of both mouse and human hearts was more thermostable in the presence of Ca2+. PMID- 14744017 TI - The putative transcriptional activator MSN1 promotes chromium accumulation in Saccharomyces cerevisiae. AB - Yeast is a good system for studying molecular mechanisms of metal tolerance. Using a mini-Tn mutagenized yeast pool, we isolated a chromate-tolerant mutant, CrT9, that displayed metal-specific tolerance since it was only tolerant to Cr(VI), not to Cr(III), Cd, As, or Fe. The Cr-tolerance of CrT9 appeared to be due to reduced Cr accumulation as it accumulated only 56% as much as WT (Y800). Using IPCR (inverse PCR), we found that the mini-Tn had been inserted at nt 741 of the transcriptional activator, MSN1. MSN1 is a multifunctional protein involved in invertase activity, iron uptake, starch degradation, pseudohyphal growth, and osmotic gene expression. We found that there was only one mini-Tn insertion in CrT9 since MSN1 and mini-Tn probes hybridized to the same DNA fragment, and the MSN1 probe detected an enlarged MSN1 mRNA. When we over expressed MSN1 in CrT9 and WT, both accumulated larger amounts of Cr. We conclude that Cr accumulation in S. cerevisiae is promoted by the transcriptional activator MSN1. PMID- 14744018 TI - Elevated P/Q type (alpha1A) and L2 type (alpha1D) Purkinje cell voltage-gated calcium channels in the cerebella of seizure prone gerbils. AB - Differences in expression of N-methyl-D-aspartate (NMDA) receptor and voltage gated Ca2+ channels (VGCC) in the gerbil cerebellum were investigated to identify routes of Ca2+ influx that may be involved in Purkinje cell damage. Immunodensities of NR1 and NR2A/B were the same in seizure resistant (SR) and seizure sensitive (SS) gerbils. However, both P/Q type (alpha1A) and L2 type (alpha1D) VGCC levels were higher in the Purkinje cells of SS gerbils than in those of SR gerbils, whereas N type (alpha1B) and L1 type (alpha1C) VGCC levels were similar in the two groups. Our findings suggest that increases in P/Q type (alpha1A) and L2 type (alpha1D) VGCC are implicated in the degeneration of Purkinje cells in SS gerbils. PMID- 14744019 TI - Age-related changes of parvalbumin immunoreactive neurons in the rat main olfactory bulb. AB - Parvalbumin (PV) is found in the olfactory system, including the main olfactory bulb, and is thought to be one of the neuroactive substances in olfaction. Changes in PV immunoreactivity in the olfactory system during aging have not been examined. We investigated such changes in the main olfactory bulb (MOB) of the rat at postnatal month 1 (PM 1), PM 3, PM 6, PM 12 and PM 24. PV-IR neurons were almost completely restricted to the external plexiform layer. At PM 1 there were only a few PV-IR neurons; at PM 3, the number of PV-IR neurons was at its greatest but they were not well developed morphologically. At PM 6, the number of PV-IR neurons was similar to that at PM 3 and they had satellite somata with well developed processes with many varicosities. By PM 12 the number of neurons and processes had declined, and by PM 24, they had fallen even further and the remaining processes had lost most of their varicosities. We conclude that age related degeneration of PV-IR neurons in the MOB may reduce calcium buffering and affect olfactory function in senile species. PMID- 14744020 TI - Prevalence of human herpesvirus-6B in Korean hematopoietic stem cell transplantation recipients. AB - Human herpesvirus-6 (HHV-6) is a major pathogen associated with diseases of recipients of hematopoietic stem cell transplants (HSCT). We have isolated HHV-6 in Korean HSCT recipients and carried out a prospective investigation of its prevalence. We obtained peripheral blood from HSCT recipients who had signs of HHV-6 infection. Cord blood mononuclear cells (CBMC) and Sup-T1 cells were used to culture the HHV-6. Indirect immunofluorescence assays (IFA), and the polymerase chain reaction (PCR) were employed to detect HHV-6. The prevalence of HHV-6 infection in HSCT recipients was calculated on the basis of the PCR results. HHV-6 was isolated from four clinical samples. After culturing the HHV-6 in CBMC, the standard strain and the four clinical isolates were propagated in Sup-T1 cells. The infected cells became grossly enlarged and multinucleate after 7-21 days. The virus was identified primarily on the basis of the morphological changes of the cultured cells, and confirmed by specific IFA with monoclonal antibody to HHV-6. HHV-6 was detected in each sample by PCR with primers specific for the major immediate early gene. Sequencing of the standard strain and PCR products confirmed identification of the HHV-6B variant. By PCR we detected 415 instances of HHV-6 in 3966 samples (14.6% of peripheral blood mononuclear cells and 6.3% of sera), and HHV-6 DNAemia was most frequent from the second to the fourth week after HSCT. PMID- 14744021 TI - Prefractionation of protein samples for proteome analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. AB - To isolate high molecular weight (HMW) or low-abundance proteins we exploited the high resolving power provided by the molecular sieves of polyacrylamide gel matrices. Rice-leaf protein extracts were applied to a single well of an SDS polyacrylamide gel with prestained molecular size markers at both ends. After electrophoresis, the gel was cut into 4 segments according to size, and each segment was ground in extraction buffer. The eluted proteins were separated from the gel matrix by centrifugation followed by acetone precipitation, and the precipitated proteins were subjected to SDS-PAGE and 2-DE. The SDS-PAGE-based prefractionation method provided non-overlapping discrete sample pools. About 27% more protein spots were detected in the fractionated samples than in the unfractionated samples, and 17% were enhanced. The improvement was especially prominent in the case of HMW proteins. Well-separated HMW proteins were analyzed by MALDI-TOF mass spectrometry. The molecular masses of the identified proteins in the > 48 kDa gel segment were distributed between 50 and 112 kDa, thus validating this prefractionation method. Identified HMW proteins with similar mass but different pI were mostly isoforms. Thus SDS-PAGE-based size prefractionation provides improved separation and detection of HMW proteins. PMID- 14744022 TI - Role of disulfide bonds in the structure and activity of human insulin. AB - Insulin contains two inter-chain disulfide bonds between the A and B chains (A7 B7 and A20-B19), and one intra-chain linkage in the A chain (A6-A11). To investigate the role of each disulfide bond in the structure, function and stability of the molecule, three des mutants of human insulin, each lacking one of the three disulfide bonds, were prepared by enzymatic conversion of refolded mini-proinsulins. Structural and biological studies of the three des mutants revealed that all three disulfide bonds are essential for the receptor binding activity of insulin, whereas the different disulfide bonds make different contributions to the overall structure of insulin. Deletion of the A20-B19 disulfide bond had the most substantial influence on the structure as indicated by loss of ordered secondary structure, increased susceptibility to proteolysis, and markedly reduced compactness. Deletion of the A6-A11 disulfide bond caused the least perturbation to the structure. In addition, different refolding efficiencies between the three des mutants suggest that the disulfide bonds are formed sequentially in the order A20-B19, A7-B7 and A6-A11 in the folding pathway of proinsulin. PMID- 14744023 TI - Induction of apoptosis and expression of cell cycle regulatory proteins in response to a phytosphingosine derivative in HaCaT human keratinocyte cells. AB - Ceramide, a compound derived from sphingomyelin, a sphingolipid precursor, affects cell functions such as growth, differentiation, cell division and apoptosis. We have shown that the phytosphingosine derivative, tetra-acetyl phytosphingosine (TAPS), inhibits the growth of HaCaT cells mainly by inducing apoptosis. In this study, we investigated its effect on the cell cycle and on cell cycle regulatory proteins. We showed by flow cytometry and staining for BrdU and phosphorylated histone H3 that the cells accumulated in S phase and arrested in G2 phase and did not divide before undergoing apoptosis. The level of the pro apoptotic regulator Bax peaked after 6 h and then returned to normal, whereas the level of the anti-apoptotic regulator Bcl-xL, which is presumably induced in order to inhibit apoptosis, started to increase at 6 h, and remained high for 24 h. Phosphorylation of Cdc2 on Tyr-15 greatly increased while p21 rose to a plateau at 8 h. Levels of p53 and Mad2 proteins were unchanged. Our observations suggest that TAPS induces apoptosis of the HaCaT cells at least in part via transient G2 arrest. PMID- 14744024 TI - Destruction of Cucumber green mottle mosaic virus by heat treatment and rapid detection of virus inactivation by RT-PCR. AB - Heat treatment is commonly used to control viral contamination of seeds. To study virus inactivation, virus was purified from seeds contaminated with Cucumber green mottle mosaic virus (CGMMV) after various heat treatments. CGMMV particles were observed to be physically disrupted by high temperature. Analysis of viral RNA revealed that the 5' and 3' termini of the genome were protected whereas regions between 2-2.5, 3.2-3.7 and 4-4.8 kb from the 5' terminus were not. Heat inactivation of virus on seeds was confirmed by RT-PCR using primers for a heat sensitive region. The RT-PCR approach developed here may prove preferable to time and labor-intensive bioassays for assessing virus heat inactivation. PMID- 14744025 TI - Remodeling of the major mouse xenoantigen, Galalpha1-3Galbeta1-4GlcNAc-R, by N acetylglucosaminyltransferase-III. AB - beta-D-Mannoside beta-1,4-N-acetylglucosaminyltransferase III (GnT-III) catalyses the attachment of an N-acetylglucosamine (GlcNAc) residue to mannose in the beta(1-4) configuration in N-glycans, and forms a bisecting GlcNAc. We have generated transgenic mice that contain the human GnT-III gene under the control of the mouse albumin enhancer/promoter [Lee et al., (2003)]. Overexpression of this gene in mice reduced the antigenicity of N-glycans to human natural antibodies, especially in the case of the alpha-Gal epitope, Galalpha1-3Galbeta1 4GlcNAc-R. Study of endothelial cells from the GnT-III transgenic mice revealed a significant reduction in antigenicity, and a dramatic decrease in both complement and natural killer cell-mediated mouse cell lysis. Changes in the enzymatic activities of other glycosyltransferases, such as alpha1,3-galactosyltransferase, and alpha-6-D-mannoside beta-1,6 N-acetylglucosaminyltransferase V, did not point to any interaction between GnT-III and these enzymes in the transgenic mice, suggesting that this approach may be useful in clinical xenotransplantation. PMID- 14744026 TI - Evidence for a conspecific relationship between two morphologically and cytologically different forms of Korean Anopheles pullus mosquito. AB - Despite the medical importance of anopheline mosquitoes as vectors of Korean vivax malaria, differentiation between Korean anopheline mosquitoes by traditional morphological taxonomic criteria is difficult. An. yatsushiroensis is the second most common Anopheles mosquito species in Korea and a possible vector of Korean vivax malaria together with An. sinensis, the predominant anopheline species. Recently, An. yatsushiroensis has been declared a synonym of An. pullus, based on comparisons of egg morphology and adult progeny, although they differ in ecology and morphology. To verify the species status of these two ambiguous forms, we established isofemale lines of Korean An. pullus and An. yatsushiroensis (An. pullus form yatsushiroensis) mosquitoes and investigated their genetic relationship by metaphase karyotype analysis, comparing the DNA sequences of rDNA internal transcribed spacer 2 (ITS2) and mitochondrial cytochrome c oxidase subunits I (COI) and II (COII), and by hybridization experiments. Two isofemale lines had differently shaped X and Y chromosomes. However reciprocal crosses between them yielded viable progeny with completely synaptic salivary gland polytene chromosomes. DNA analyses also strongly supported their conspecificity. The two strains also showed great sequence similarity in the ITS2, COI and COII regions (variation rate = 0.0 to 0.8%). Based on these findings, we conclude that the two forms, though differing distinctly in morphological, cytological and ecological traits, remain interfertile. PMID- 14744027 TI - Purification and characterization of a cytosolic phospholipase A2 from rat liver. AB - A cytosolic phospholipase A2 (PLA2) was purified 640-fold from rat liver by sequential anion-exchange chromatography, Ca2+-precipitation/KCl-solubilization, gel filtration chromatography, and affinity chromatography. A single peak of PLA2 activity was eluted at an apparent molecular mass of 197 kDa from a Superdex 200HR gel filtration column. In the presence of Ca2+, the purified enzyme catalyzed the hydrolysis of 81.8 nmol of phosphatidylethanolamine per hour per mg of protein. The apparent Km was 1.83 nM. The enzyme was inhibited by arachidonyl trifluoromethyl ketone (AACOCF3), an inhibitor of cPLA2. However, it was not inhibited by bromoenol lactone (BEL), an inhibitor of iPLA2, and p-bromophenacyl bromide (p-BPB), an inhibitor of sPLA2. These data suggest that the purified enzyme is a novel Ca2+-dependent cytosolic PLA2. PMID- 14744028 TI - Reduction of the Gal-alpha1,3-Gal epitope of mouse endothelial cells by transfection with the N-acetylglucosaminyltransferase III gene. AB - In order to prevent hyperacute rejection in pig-to-human xenotransplantation, it would be very useful to be able to down-regulate the Gal alpha1-3 Galbeta 1-4 GlcNAc-R (alpha-Gal epitope) in mouse and swine tissues. When the beta-D mannoside beta-1,4-N-acetylglucosaminyl-transferase III (GnT-III) gene was introduced into mouse aorta endothelial cells (MEC) their susceptibility to complement-mediated cell lysis by normal human serum (NHS) was reduced. Expression of GnT-III also suppressed the antigenicity of MEC to human natural antibodies as shown by binding of Griffonia simplicifolia 1 isolectin (GS1B4 lectin) to the alpha-Gal epitope. Western blot analysis indicated that the reactivity of the glycoproteins of the transfectants to NHS and GSIB4 lectin was reduced to approximately the same extent. Thus GnT-III, a key enzyme involved in the formation of branched N-linked sugars, reduces the expression of xenoantigens, suggesting that this approach may be of value in clinical xenotransplantation. PMID- 14744029 TI - The effects of ethidium bromide and Mg++ ion on strand exchange in the Hin mediated inversion reaction. AB - The biochemical reaction of a site-specific recombinase such as Hin invertase or gammadelta resolvase starts with binding of the recombinase to its recombination site and cleavage of the DNA in the center of the site. This is followed by strand exchange and finally ligation of the ends of the recombined strands. Previous biochemical studies have shown that Hin invertase and gammadelta resolvase cannot proceed beyond DNA cleavage in the absence of Mg++ ion, indicating that these recombinases require Mg++ ion in the strand exchange process. We have observed that the intercalating agent, ethidium bromide (2 microM), does not interfere with DNA cleavage, but slows strand exchange in a concentration-dependent manner. Levels of Mg++ ion below 5 mM also slow strand exchange substantially. We infer that random intercalation of ethidium bromide inhibits unwinding of the double helix at the recombination site in the negatively supercoiled DNA and propose that Mg+ may be required for Hin to deform the secondary structure of B-DNA prior to strand exchange. PMID- 14744030 TI - Enhanced uptake of a heterologous protein with an HIV-1 Tat protein transduction domains (PTD) at both termini. AB - Poor membrane permeability of proteins is a major limitation of protein therapy. In a previous study, we showed that the minimal sequence required for efficient transduction of Tat-GFP is the basic domain from 49-57 of HIV-1 Tat called the protein transduction domain (PTD. Here we have generated HIV-1 Tat PTD GFP fusion proteins in which HIV-1 Tat PTD is fused with the N- and/or C-termini of GFP. The various GFP fusion proteins were purified from Escherichia coli and characterized for their ability to enter mammalian cells using Western blot analysis, confocal microscopy and flow cytometry. The GFP fusion protein with Tat PTD at its C terminus was taken up as efficiently as the GFP fusion protein with Tat PTD at its N-terminus. However, the same protein with PTDs at its both termini was taken up even more efficiently. All the GFP fusion proteins were present in both the nucleus and cytosol of the transduced cells. Uptake was lower at 4 degrees C than at 37 degrees C. The availability of the expression vectors developed in this study may help to devise novel strategies in the rational development of protein based drugs. PMID- 14744031 TI - Expression of Rbp9 during mid-oogenesis induces apoptosis in egg chambers. AB - The Drosophila RNA binding protein RBP9 is expressed in both ovarian germline cells and neuronal cells of the adult central nervous system. Expression is limited to postmitotic cells in both cases. It has been suggested that Rbp9 is involved in the regulation of genes that function in cell proliferation or differentiation. We examined the effect of ectopic expression of Rbp9 in germline and somatic cells using the Gal4 system. Over-expression of Rbp9 in various tissues caused severe developmental defects. In particular, expression during mid oogenesis using nos-Gal4:VP16 caused apoptosis in stage 10 egg chambers. A similar phenotype has been reported when one of the effector caspases of Drosophila, Dcp-1, is ectopically expressed during mid-oogenesis with the same Gal4 driver. Tight control of Rbp9 transcription seems to be critical for the proper development of Drosophila. PMID- 14744032 TI - Cloning of the correct full length cDNA of NF-kappaB-repressing factor. AB - NF-kappaB-repression factor (NRF) is a nuclear inhibitor of NF-kappaB proteins that can silence the IFNbeta promoter. Since NRF was cloned in 1999, in-depth studies have been conducted on the biological functions of this constitutive repressor of NF-kappaB proteins. During large-scale sequencing of a human fetal brain cDNA library we isolated a novel human cDNA that proved to be a correct full-length NRF cDNA. The deduced protein contains 690 aa, and has a G-patch and an R3H domain at its C-terminus. The size of the protein is consistent with its counterparts in mouse and rat. There is considerable evidence that there are some mistakes in the NRF cDNA sequence reported by Nourbakhsh. Here we report the correct, full-length cDNA and protein sequences of NRF. Full-length NRF cDNA is 3247 bp long, contains three exons and maps to human chromosome Xq24. RT-PCR shows that NRF is widely expressed in human tissues. PMID- 14744033 TI - Ginsenosides enhance the transduction of tat-superoxide dismutase into mammalian cells and skin. AB - We previously reported that Tat-Cu,Zn-superoxide dismutase (Tat-SOD), a major antioxidant enzyme, can be directly transduced into mammalian cells and skin [Kwon et al. (2000); Park et al. (2002)]. To enhance the therapeutic potential of Tat-SOD in the treatment of various disorders, we screened a number of natural products for their ability to increase transduction efficiency. Ginsenosides were effective with cultured HeLa cells and enhanced the penetration of Tat-SOD into both the epidermis and the dermis of the subcutaneous layer when sprayed on mice skin. Although their mechanism of action is not fully understood we believe that ginsenosides may be useful cofactors with this antioxidant enzyme in anti-aging cosmetics or as a therapeutic protein in disorders related to reactive-oxygen species. PMID- 14744034 TI - Nucleotide sequences of two Korean isolates of Cucumber green mottle mosaic virus. AB - The nucleotide sequences of the genomic RNAs of Cucumber green mottle mosaic virus Korean watermelon isolate (CGMMV-KW) and Korean oriental melon isolate (CGMMV-KOM) were determined and compared to the sequences of other tobamoviruses including CGMMV strains W and SH. Each CGMMV isolate had a genome of 6,424 nucleotides. Each also had 60 and 176 nucleotides of 5' and 3' untranslated regions (UTRs), respectively, and four open reading frames (ORF1-4). ORFs 1 to 4 encode proteins of 129, 186, 29, and 17.4 kDa, respectively. The nucleotide and deduced amino acid sequences of CGMMV-KOM and CGMMV-KW were more than 98.3% identical. When compared to other CGMMV strains in a phylogenetic analysis they were found to form a distinct virus clade, and were more distantly related to other tobamoviruses (23.5-56.7% identity). PMID- 14744035 TI - RNA aptamers that bind the nucleocapsid protein contain pseudoknots. AB - We screened two independent RNA libraries consisting of molecules of 50 nucleotides of random sequence, one of which had additional viral psi-sequences to isolate RNA aptamers that bound to the mature form of the nucleocapsid (NC) protein of Human Immunodeficiency Virus Type-1 (HIV-1). Surface Plasmon Resonance measurements and gel shift assays showed that the RNA aptamers bound with high affinity and specificity. We employed RNase footprinting to characterize the RNA structures and to map their protein binding sites. Most of the selected RNA aptamers contained a plausible pseudoknot in addition to the characteristic stem loop structure. Moreover, the pseudoknots were part of the NC binding sites. We propose that higher order structures such as pseudoknots may constitute binding motifs for nucleic acid binding proteins, especially for NC protein, which is a nucleic acid chaperone. PMID- 14744036 TI - Neuroimaging in eating disorders. AB - Neuroanatomical and functional studies in the eating disorders (ED) are reviewed. Typically, anorexia nervosa (AN) is associated with cerebral spinal fluid spaces enlargement which generally recover as a function of re-feeding. However, specific cortical areas fail to correct in weight restored anorectic patients suggesting trait-related abnormalities. Functional changes in AN associated with starvation reverse with weight recovery, however, reduced 5-HT2A receptor binding may be fundamental to the pathophysiology of AN since this remains after long term weight restoration. Structural studies of bulimia nervosa (BN) provide evidence of brain atrophy, in the absence of significant weight loss but potentially related to chronic dietary restriction. Functional investigations reveal reduced thalamic and hypothalamic serotonin transporter availability in BN which increases with longer illness duration. Thus, BN is associated with substantial structural and functional alterations despite normal weight. Recent advances in neuroimaging techniques and their interpretation are increasing our understanding of normal processes in the control of food intake including neuroanatomical correlates of hunger and satiety. Taken together with the structural and functional changes observed in the ED, neuroimaging provides a powerful platform to identify the underlying trait-related pathophysiological mechanisms in the aetiology and maintenance of AN and BN. PMID- 14744037 TI - Auditory brainstem evoked response in juvenile rats fed rat milk formulas with high docosahexaenoic acid. AB - Previous studies found that juvenile offspring of rats fed high docosahexaenoic acid (DHA; 22:6n-3) diets through gestation and lactation had longer auditory brainstem-evoked response (ABR) accompanied by higher 22:6n-3 and lower arachidonic acid (ARA; 20:4n-6) in brain. In the present study, ABR was assessed in juvenile rats fed high-DHA diets only postnatally. METHODS: Rat pups were fed rat milk formulas with varying amounts of DHA and ARA to 19 days of age followed by diets with the corresponding fatty acids. The high-DHA group was fed 2.3% of fatty acids as DHA, the DHA + ARA group was fed DHA and ARA at 0.6 and 0.4% of fatty acids, levels similar to those in some infant formulas, and the unsupplemented group was fed no DHA or ARA. ABR and fatty acid and monoamine levels in brain were measured on postnatal days 26-28. Statistical analyses were measured by ANOVA. RESULTS: ARA and DHA levels in brain increased with supplementation. ABR was shorter in the high-DHA group than the DHA + ARA group and not different from the unsupplemented or dam-reared suckling group. Norepinephrine levels in the inferior colliculus were lower in the high-DHA group than the DHA + ARA group and higher in all formula groups compared to the dam reared group. CONCLUSION: In contrast to the longer ABR in juvenile offspring of rats fed high-DHA through gestation and lactation, ABR was shorter in juvenile rats fed high-DHA diets only after birth than rats fed ARA + DHA. Further studies are needed to understand the relationship between dietary DHA, norepinephrine, and auditory system development over a range of DHA intakes and discrete periods of development. PMID- 14744038 TI - Dietary n-3 and n-6 fatty acids alter avian pituitary sensitivity. AB - The effects of dietary saturated and polyunsaturated fatty acids (PUFAs) of the n 3 and n-6 series on avian pituitary sensitivity were investigated by infusing human growth hormone (GH) releasing hormone--fragment 1-29--and chicken luteinising hormone releasing hormone (LHRH) into catheterized broiler chickens. At 3 weeks of age three groups (n = 18; six birds per group) were fed for 6 weeks isonitrogenous and isoenergetic experimental diets containing 80 g/kg of edible tallow (saturated fatty acids), fish oil (n-3 PUFAs) or sunflower oil (n-6 PUFAs). Jugular catheterisation was performed under general anaesthesia during week four of the dietary treatments and the birds allowed 7 days post surgery to recover. A bolus of LHRH (20 microg/bird) and a GH releasing hormone (12.5 microg/kg) infusion was given on different days to each chicken and serial blood samples taken over a 1 h period. Plasma luteinising hormone and GH concentrations were measured by radioimmunoassay. Pre-infusion GH concentrations were similar for the tallow, fish and sunflower oil dietary groups (5.2 +/- 3.9, 5.2 +/- 1.0 and 6.1 +/- 3.1 ng/ml, respectively), however, GH concentration in response to the GH releasing hormone infusion was elevated in the sunflower oil group (44.7 +/- 5.7 ng/ml) when compared to chicken fed tallow (33.7 +/- 9.7ng/ml) or fish oil (21.3 +/- 5.0 ng/ml). There was a significant decrease (P < 0.05) in the clearance rate of plasma GH for the birds fed the fish oil compared with those fed sunflower oil with an intermediate value being observed in the tallow fed group. Pre-infusion plasma luteinising hormone concentrations for the birds fed tallow (3.2 +/- 0.7 ng/ml) were significantly elevated (P < 0.05) when compared to birds fed either the sunflower oil (0.84 +/- 0.25 ng.ml) or fish oil (0.93 +/- 0.22 ng/ml) diets. There were no significant differences between the dietary groups in either the maximal plasma luteinising concentration or its disappearance rate following the LHRH infusion. The data demonstrate that dietary fatty acids alter avian pituitary sensitivity and this modulation is determined by the nature of the dietary fat rather than the degree of saturation per se. In addition, this study also shows that dietary fats have a differential effect on pituitary cell activity and are specific to certain pituitary cell types. PMID- 14744039 TI - Effects of long-term vitamin K (phylloquinone) intake on retina aging. AB - Emerging evidence suggests neuroprotective functions of vitamin K and/or vitamin K-dependent proteins. We investigated the effect of dietary vitamin K on retina aging (thinning). Female Sprague-Dawley rats were maintained from weaning on low (80 microg kg(-1) diet), adequate (500 microg kg(-1) diet) or high (2000 microg kg(-1) diet) levels of vitamin K1 (phylloquinone). Relative concentrations of brain vitamin K associated with these diets were 1: 3.3: 25 (K1) and 1: 2.7: 9.0 (menaquinone-4). Histomorphometry of old (21 month) rats revealed positive associations between vitamin K and thickness of retina layers, especially in the equatorial/peripheral retina. No association of diet and retina thickness was detected among young (6 month) animals. The sparing effect of vitamin K in the retina was most evident in the inner plexiform layer and in the photoreceptor inner and outer segments. Surprisingly, we observed no effect of vitamin K on the age-dependent loss of photoreceptor cells, interneurons or ganglion cells. These data suggest a role for vitamin K in maintaining the aging retina and suggest that the sparing effect of vitamin K does not reflect the survival-promoting (anti-apoptotic) activities of vitamin K-dependent proteins. PMID- 14744040 TI - Low dietary protein is associated with an increase in food intake and a decrease in the in vitro release of radiolabeled glutamate and GABA from the lateral hypothalamus. AB - Moderately low-protein diets lead to a rapid increase in food intake and body fat. The increase in feeding is associated with a decrease in the concentration of serum urea nitrogen, suggesting that the low-protein-induced increase in food intake may be related to the decreased metabolism of nitrogen from amino acids. We hypothesized that low dietary protein would be associated with a decrease in the synaptic release of two nitrogen-containing neurotransmitters, GABA and glutamate, whose nitrogen can be derived from amino acids. In this study, we examined the effects of a low-protein diet (10% casein) in Sprague-Dawley rats on the in vitro release of 3H-GABA and 14C-glutamate from the lateral and medial hypothalamus. The low-protein diet increased food intake by about 25% after one day. After four days, the in vitro release of radiolabeled GABA and glutamate was assessed. The calcium-dependent, potassium-stimulated release of radiolabeled GABA and glutamate from the lateral hypothalamus was decreased in rats fed the low-protein diet. The magnitude of neurotransmitter release from the lateral hypothalamus inversely correlated with food intake. No dietary differences in the release of neurotransmitters from the medial hypothalamus were observed. These results support the contention that alterations in nitrogen metabolism are associated with low-protein-induced feeding. PMID- 14744041 TI - Quantitative genetic analysis of ventral midbrain and liver iron in BXD recombinant inbred mice. AB - Male and female mice from 15 of the BXD/Ty recombinant inbred strain panel were examined for regional brain and liver iron content. Brain regions included medial prefrontal cortex, nucleus accumbens, caudate-putamen and ventral midbrain. Our focal tissue was the ventral midbrain, containing the ventral tegmentum and substantia nigra. This area contains the perikarya of the dopamine neurons that project to nucleus accumbens and caudate-putamen. Genetic correlations between ventral midbrain and liver iron content were not statistically significant, suggesting that peripheral and central iron regulatory systems are largely independent. Correlations between ventral midbrain iron and iron in the caudate putamen and nucleus accumbens, but not the prefrontal cortex were moderately high and significant. Ventral midbrain and liver iron contents were subjected to quantitative trait loci analysis to identify associated chromosomal locations. This analysis revealed several suggestive loci for iron content in ventral midbrain but fewer loci for liver. Genetic correlations between ventral midbrain iron and published dopamine functional indices were significant, suggesting a link between ventral midbrain iron status and central dopamine neurobiology. This work shows the value of quantitative genetic analysis in the neurobiology of iron and in showing the close association between ventral midbrain iron and nigrostriatal/mesolimbic dopamine function. PMID- 14744042 TI - Spatial learning deficits induced by muscimol and CL218,872: lack of effect of prenatal malnutrition. AB - The sensitivity of prenatal protein malnourished rats to the amnestic properties of the direct GABAA receptor agonist muscimol and the selective benzodiazepine (BZ) receptor agonist, CL218,872, was studied in the male offspring of rats provided with a protein deficient diet (6% casein) for 5 weeks prior to mating and throughout pregnancy. At postnatal day 90, rats were tested during acquisition of the submerged platform version of the Morris water maze task using four systemic doses of muscimol (0.1, 0.3, 1.0 and 1.8 mg/kg i.p.) or three systemic doses of CL218,872 (1.0, 3.2, and 5.6 mg/kg i.p.). In a dose dependent manner both drugs impaired acquisition of the task and impaired accuracy of the search pattern on the probe trial (platform removed). However, neither drug dissociated the performance of the two nutritional groups. These data are important in light of previous findings of differential behavioral effects of the non-specific BZ agonist, chlordiazepoxide (CDP), on spatial learning and on drug discrimination in prenatally malnourished rats and in the context of previous findings of reduced sensitivity to the anxiolytic effects of non-specific BZ receptor agonists across a wide variety of models of malnutrition. The present findings also support the concept that prenatal malnutrition does not affect the global functioning of the GABAA receptor, but fundamentally alters the way in which a subset of GABAA receptors (i.e. those containing the alpha2, alpha3 and/or the alpha5 but not the alpha1 subunit) is modulated by BZs. PMID- 14744043 TI - Determination of fatty acid levels in erythrocyte membranes of patients with chronic fatigue syndrome. AB - Chronic fatigue syndrome (CFS) is an illness characterized by persistent and relapsing fatigue, often accompanied by numerous symptoms involving various systems of whole body. The etiology of CFS remains unclear. Literature reported whether the concentrations of the essential fatty acids in red cell membranes of CFS patients were decreased is controversial. In our study, Forty-two patients who fulfilled the diagnostic criteria defined by Centers for Disease Control and Prevention (CDC). Thirty-seven age- and sex-matched controls were selected from healthy medical staffs and volunteers. After lipid analysis, we found that the levels of the arachidonic acid (ARA) and docosahexanoic acid (DHA) were decreased in patients suffered from CFS. However, the levels of the palmitic acid and oleic acid were increased. We speculated that there are two possible mechanisms--one of which is that oxidative stress has led to an excessive oxidation and resulting in the above fatty acids. Alternatively, insufficiency of ingestion of fatty acids might not be the major cause. PMID- 14744044 TI - Effects of blood flow on skin heating induced by millimeter wave irradiation in humans. AB - We have previously reported species differences in the rate of skin heating in response to millimeter wavelength microwave exposure. We hypothesized that these differences were predominantly a function of species differences in the ability to increase skin blood flow during local heating. Mathematical modeling also suggested that, in humans, the rate of skin heating during prolonged millimeter wavelength exposure would be dependent on skin blood flow. In order to empirically test this hypothesis, we determined the role of baseline skin blood flow on the rate of cutaneous heating induced by 94-GHz microwave energy in humans (3 female, 3 male) using infrared thermography and laser Doppler imaging to measure skin temperature and relative skin blood flow, respectively. Millimeter wavelength exposure intensities used were high power (HP), 1 W x cm( 2) for 4 s and low power, 175 mW cm(-2) for 180 s. Skin blood flow was (a) normal, (b) eliminated using a blood pressure cuff to occlude forearm blood flow, or (c) elevated by heating the skin prior to irradiation. Results showed that for the HP exposures, these manipulations did not influence the rate of skin heating. For the low power exposures, occlusion of baseline skin blood flow had a small impact on the subsequent rate of heating. In contrast, a two-fold elevation in baseline skin blood flow had a profound impact on the subsequent rate of heating, resulting in a substantially lower rate of heating. Occlusion of an elevated skin blood flow reversed this lower rate of heating. The results of these studies demonstrate that relatively small changes in skin blood flow may produce substantial alterations in the rate of skin heating during prolonged 94-GHz exposure. PMID- 14744045 TI - Dose reconstruction by EPR spectroscopy of tooth enamel: application to the population of Zaborie village exposed to high radioactive contamination after the Chernobyl accident. AB - Individual irradiation doses were determined by electron paramagnetic resonance spectroscopy of the tooth enamel of the inhabitants of Zaborie, the most contaminated inhabited settlement not evacuated after the Chernobyl accident. Dose determination was performed using a specially developed automatic spectrum processing procedure. Spectrum processing was carried out in different operating modes, and average results were taken in order to reduce the contribution of uncertainty in dose determination caused by spectrum processing. The absorbed doses determined in enamel were corrected to take into account the contribution of natural background radiation and to determine the individual excess dose due to radioactive contamination of the territory. Individual excess doses are compared to calculated individualized doses to teeth, estimated using the local radioactive contamination levels, dose rates, and information concerning individual behavior. The individual excess doses measured by electron paramagnetic resonance spectroscopy and the calculated individualized doses are fully independent. Mean square variation between results of two methods was found to be 34 mGy, which is consistent with error estimation for both methods. This result can validate both the methodology of signal processing presented here when using electron paramagnetic resonance dosimetry of tooth enamel for low doses and the methodology of individualized dose calculation. PMID- 14744046 TI - Application of NCRP air screening factors for evaluating both routine and episodic radionuclide releases to the atmosphere. AB - Two separate methods were used to identify the most important historic airborne releases of radionuclides at the Idaho National Engineering and Environmental Laboratory (INEEL) with regard to potential human health impact. Both routine and episodic releases were evaluated. Although not specifically intended for an initial screening or ranking evaluation, particularly for episodic releases, the National Council on Radiation Protection and Measurements (NCRP) screening method was shown to be a valid method for providing a measure of the relative importance of both routine and episodic radionuclide releases, based on comparisons with the Radiological Safety Analysis Code (RSAC). For the work at the INEEL, a relative ranking procedure was used to identify the most important releases because a screening criterion (e.g., dose or risk value) against which the potential health impacts of the releases could be measured was not established. In addition, a precedent for a screening-level evaluation of episodic releases is lacking at this time. As a result, a ranking procedure was considered necessary because it was not clear that the NCRP method would provide screening-level dose estimates for episodic releases that could be defensibly compared to a screening criterion. To evaluate the NCRP method at the INEEL, routine operational releases were evaluated and ranked separately from episodic, or acute, releases because different assumptions and approaches were required to assess their potential importance. Based on comparisons with the RSAC method, the NCRP method may slightly underpredict the ingestion dose for episodic releases; however, using the NCRP screening method to identify the relative importance of release events, radionuclides, years, and facilities was shown to be valid and defensible for both routine and episodic releases. Because of the NCRP method's simplicity and relative ease of application, it provides a cost-effective and scientifically defensible way to make decisions and set priorities about decisions and directions in risk assessment. PMID- 14744047 TI - Concurrent determination of 224Ra, 226Ra, 228Ra, and unsupported 212Pb in a single analysis for drinking water and wastewater: dissolved and suspended fractions. AB - A technique has been developed for the measurement of 224Ra, 226Ra, 228Ra, and unsupported 2t2Pb concurrently in a single analysis. The procedure can be applied to both drinking water and wastewater, including the dissolved and suspended fractions of a sample. For drinking water samples, using 3-L aliquots, the radium isotopes are isolated by a fast PbSO4 co-precipitation and then quantified by gamma-ray spectroscopy. The radium isotopes 224Ra, 226Ra, and 228Ra are measured through their gamma-ray-emitting decay products, 212Pb, 214Pb (and/or 214Bi), and 228Ac, respectively. Because of the short half-life of 224Ra (T1/2 = 3.66 d), the precipitate should be counted within 4 d of the sample collection date. In case the measurement of unsupported 212Pb (T1/2 = 10.64 h) is required, the gamma-ray analysis should be initiated as soon as possible, preferably on the same day of collection. The counting is repeated after about 21 d to ensure the 226Ra progeny are in equilibrium with their parent. At this point, the 228Ac equilibration with its 228Ra parent is already established. In the case of samples containing suspended materials, an aliquot of sample is filtered and then the filtrate is treated as described above for drinking water samples. The suspended fraction of sample, collected on the filter, is directly analyzed by gamma-ray spectroscopy with no further chemical separation. Aliquots of de-ionized water spiked with various radium standards were analyzed to check the accuracy and precision of the method. In addition, analysis results of actual samples using this method were compared with the ones performed using U.S. Environmental Protection Agency approved procedures, and the measured values were in close agreement. This method simplifies the analytical procedures and reduces the labor while achieving the precision, accuracy, and minimum detection concentration requirements of EPA's Regulations. PMID- 14744048 TI - Radon concentrations in some dwellings of Tunisia. AB - Indoor air radon concentrations are still unknown in Tunisia. For the first time, they have been determined in several regions of the country using open alpha track dosimeters containing LR-115 film. Measurements were taken in 69 dwellings located around greater Tunis during 1 y, changing dosimeters every 2 mo. In 12 other locations, devices were placed during 2 winter months. The median of 1,217 measurements was 40 Bq m(-3) and 93.4% of them were less than 100 Bq m(-3). The highest concentration was 392 Bq m(-3). In Tunis, concentrations were higher during winter. Indoor air radon figures varied with geographic location: the highest values were found in Jendouba, Gafsa, Beja, and Tataouine government districts where phosphate and lead mines and deposits are present. This first study showed that indoor air radon concentrations are low in Tunisia, but further studies should be performed in localized areas, taking into consideration the geology, the climatic variations, and the building material. PMID- 14744049 TI - Dosimetry of metal tritide particles as evaluated by the ICRP 66 model and a biokinetic model from laboratory rats. AB - Internal radiation from inhalation of metal tritide aerosols may present a significant radiation protection problem for nuclear facility workers. Dose was evaluated for three metal tritides: hafnium, titanium, and zirconium. The study included in vitro and in vivo exposures. The inhalation doses for the three materials were calculated by using the ICRP 66 lung model. The doses also were evaluated by a biokinetic model, which was developed according to the results of animal studies. Results showed that the hafnium tritide particles were the most dense but they had a lower dissolution rate and a higher retention rate. Among these three tritides, hafnium was classified as a Type S material according to the ICRP 66 publication, whereas titanium and zirconium ranked between Type M and F materials. The dissolution rate of hafnium tritide appeared to agree well with the in vitro and in vivo studies. The dissolution rates of the other two materials in the in vitro studies were a little higher than those of the in vivo studies. The doses calculated by the ICRP 66 model for all materials were approximately two orders smaller than the doses obtained by the animal studies. This bias was caused by the different intake methods of the ICRP 66 model (inhalation) and in the animal study (instillation). The doses were on the same order while correcting for deposition fractions. The effective doses for hafnium, titanium, and zirconium tritides were 5.43 x 10(-10), 9.05 x 10(-11), and 6.5 x 10(-10) Sv Bq(-1), respectively, according to the animal studies. PMID- 14744050 TI - Modeling of an industrial environment: external dose calculations based on Monte Carlo simulations of photon transport. AB - External gamma exposures from radionuclides deposited on surfaces usually result in the major contribution to the total dose to the public living in urban industrial environments. The aim of the paper is to give an example for a calculation of the collective and averted collective dose due to the contamination and decontamination of deposition surfaces in a complex environment based on the results of Monte Carlo simulations. The shielding effects of the structures in complex and realistic industrial environments (where productive and/or commercial activity is carried out) were computed by the use of Monte Carlo method. Several types of deposition areas (walls, roofs, windows, streets, lawn) were considered. Moreover, this paper gives a summary about the time dependence of the source strengths relative to a reference surface and a short overview about the mechanical and chemical intervention techniques which can be applied in this area. An exposure scenario was designed based on a survey of average German and Hungarian supermarkets. In the first part of the paper the air kermas per photon per unit area due to each specific deposition area contaminated by 137Cs were determined at several arbitrary locations in the whole environment relative to a reference value of 8.39 x 10(-4) pGy per gamma m(-2). The calculations provide the possibility to assess the whole contribution of a specific deposition area to the collective dose, separately. According to the current results, the roof and the paved area contribute the most part (approximately 92%) to the total dose in the first year taking into account the relative contamination of the deposition areas. When integrating over 10 or 50 y, these two surfaces remain the most important contributors as well but the ratio will increasingly be shifted in favor of the roof. The decontamination of the roof and the paved area results in about 80-90% of the total averted collective dose in each calculated time period (1, 10, 50 y). PMID- 14744051 TI - Use of the VIP-Man model to calculate energy imparted and effective dose for x ray examinations. AB - A male human tomographic model was used to calculate values of energy imparted (epsilon) and effective dose (E) for monoenergetic photons (30-150 keV) in radiographic examinations. Energy deposition in the organs and tissues of the human phantom were obtained using Monte Carlo simulations. Values of E/epsilon were obtained for three common projections [anterior-posterior (AP), posterior anterior (PA), and lateral (LAT)] of the head, cervical spine, chest, and abdomen, respectively. For head radiographs, all three projections yielded similar E/epsilon values. At 30 keV, the value of E/epsilon was approximately 1.6 mSv J(-1), which is increased to approximately 7 mSv J(-1) for 150 keV photons. The AP cervical spine was the only projection investigated where the value of E/epsilon decreased with increasing photon energy. Above 70 keV, cervical spine E/epsilon values showed little energy dependence and ranged between approximately 8.5 mSv J(-1) for PA projections and approximately 17 mSv J(-1) for AP projections. The values of E/epsilon for AP chest examinations showed very little variation with photon energy, and had values of approximately 23 mSv J(-1). Values of E/epsilon for PA and LAT chest projections were substantially lower than the AP projections and increased with increasing photon energy. For abdominal radiographs, differences between the PA and LAT projections were very small. All abdomen projections showed an increase in the E/epsilon ratio with increasing photon energy, and reached a maximum value of approximately 13.5 mSv J(-1) for AP projections, and approximately 9.5 mSv J(-1) for PA/lateral projections. These monoenergetic E/epsilon values can generate values of E/epsilon for any x-ray spectrum, and can be used to convert values of energy imparted into effective dose for patients undergoing common head and body radiological examinations. PMID- 14744052 TI - Theoretical organically bound tritium dose estimates. AB - This paper illustrates a theoretical approach to estimating the dose associated with the ingestion of both organically bound tritium and tissue free water tritium relative to the ingestion of only tissue free water tritium. Organically bound tritium, specifically non-exchangeable OBT, can result in an increased dose relative to that from exchangeable organically bound tritium and tissue free water tritium because of the longer biological half-life of the former resulting in a dose conversion factor that is twice that of the latter. Non-exchangeable organically bound tritium is tritium that is bound to carbon whereas exchangeable organically bound tritium is tritium bound to oxygen, nitrogen, or sulfur. Tissue free water ranges from 85+% in most fruits and vegetables down to approximately 10% in grains. The remaining edible food mass consists, in part, of exchangeable and nonexchangeable hydrogen that is incorporated into carbohydrates, proteins and fat. The potential organically bound tritium content of several common food items was calculated knowing the amount of bound and unbound hydrogen that exists in these foods and by assuming that the hydrogen to tritium ratio is the same for the "free water" and bound hydrogen compartments. The theoretical ratio of dose from ingestion of organically bound tritium and tissue free water tritium to dose from ingestion of only tissue free water tritium was calculated to be on average within 12%, 30%, and 261% of experimentally based values for fruits and vegetables, meats and eggs, and grains, respectively. The difference is attributed to the T:H ratio being a function of the kinetics associated with the assimilation of tritium into the tissues. PMID- 14744053 TI - Assessment of the current internal dose due to 137Cs and 90Sr for people living within the Semipalatinsk Test Site, Kazakhstan. AB - The Semipalatinsk Test Site in Kazakhstan was one of the major sites used by the USSR for testing nuclear weapons for more than 40 y. Since the early 1990's, responsibility for the site has passed to the Kazakh authorities. There has been a gradual re-establishment of agricultural use such as horse and sheep farming. Therefore, it has become important to evaluate the current and future risk to people living on and using the contaminated area. Internal dose assessment is one of the main components of the total dose when deriving risk factors for population living within the test site. Internal doses based on food monitoring and whole body measurements were calculated for adults and are in the range of 13 500 microSv y(-1) due to radiocesium and radiostrontium. PMID- 14744054 TI - Strontium biokinetics in humans: influence of alginate on the uptake of ingested strontium. AB - Radioactive isotopes of strontium, mainly 90Sr, released into the environment due to nuclear accidents may contribute significantly to the internal radiation exposure of members of the public after ingestion of strontium with contaminated foodstuffs. The committed radiation dose is significantly dependent on the fraction of the ingested activity that crossed the gut wall (f1 value). In tracer kinetic investigations, the effect of sodium alginate on the gastrointestinal absorption of strontium was studied in human subjects. Sodium alginate was proven to be a potent agent for reducing strontium absorption with high efficiency and virtually no toxicity. The data obtained show that the uptake of ingested strontium from milk was reduced by a factor of nine when alginate was added to milk. It is concluded that alginate preparations are a suitable antidote against radiostrontium. PMID- 14744055 TI - An analysis of 45 years of reported overexposure incidents in Texas, 1956 to 2001. AB - Sources of ionizing radiation are commonly encountered in a wide variety of modern work settings. The controls in place to ensure the safe use of these sources have proven to be quite effective, as events involving occupational doses in excess of established limits are quite rare. Nonetheless, instances of doses in excess of established limits, commonly referred to as "overexposures," do occur, but the rarity of such events has resulted in a body of scientific knowledge that consists essentially of sporadic case reports. In this study, incident reports describing radiation overexposure events recorded in Texas from the years 1956 to 2001 were obtained and recorded into a computerized database using a pre-established set of codes. The data were then analyzed for the identification of possible trends or commonalties. During the 45-y period of study, overexposure events accounted for 50% (n = 3,796) of all the radiation related incidents recorded in Texas for the time period (n = 7,534). Of the overexposure events, 65% (n = 2,342) resulted in the actual deposition of energy in the individual exposed. The remainder were determined to be doses recorded only by a personal dosimetry device. In most of the cases where doses were actually delivered to an individual, the doses were less than 0.05 Sv (5 rem). In only 0.5% of the cases (n = 13) were doses greater than 1 Sv (100 rem). The predominant sources reported as involved in the events included 192Ir, 60Co, and 137Cs. The information derived from the analysis may serve as a basis for a variety of interventions, such as preventative education activities, regulatory modifications, and the possible re-design of equipment identified as commonly associated with such events. The results of the study can also assist in the training of health care providers, as the recognition of common causes and sources of overexposures and subsequent treatments can be forecasted and summary treatment protocols developed. PMID- 14744056 TI - The Puskin observation on smoking as a confounder in ecologic correlations of cancer mortality rates with average county radon levels. PMID- 14744057 TI - Annual ICRP meeting. PMID- 14744058 TI - Energy employees occupations illness compensation program. PMID- 14744059 TI - High level cesium source secured in Cote d'Ivoire. PMID- 14744060 TI - First U.S.-Russian business venture in closed nuclear city. PMID- 14744061 TI - ORS interview with Sue Dupre, RSO from Princeton University. PMID- 14744062 TI - Health physics ethics. AB - Ethics is defined in the New World Dictionary as ". . . moral principles governing appropriate conduct for an individual or group." The Health Physics Membership Directory contains 2 references for professional conduct for health physicists. The first is for members of the Health Physics Society. The second is for Certified Health Physicists. They are similar: A health physicist must always maintain the highest ethical standards whether beginning a career or having practiced for decades. A review of some key principles by example will hopefully demonstrate how to avoid ethical dilemmas for health physicists. PMID- 14744063 TI - Considerations for applying VARSKIN mod 2 to skin dose calculations averaged over 10 cm2. AB - VARSKIN Mod 2 is a DOS-based computer program that calculates the dose to skin from beta and gamma contamination either directly on skin or on material in contact with skin. The default area for calculating the dose is 1 cm2. Recently, the U.S. Nuclear Regulatory Commission issued new guidelines for calculating shallow dose equivalent from skin contamination that requires the dose be averaged over 10 cm2. VARSKIN Mod 2 was not filly designed to calculate beta or gamma dose estimates averaged over 10 cm2, even though the program allows the user to calculate doses averaged over 10 cm2. This article explains why VARSKIN Mod 2 overestimates the beta dose when applied to 10 cm2 areas, describes a manual method for correcting the overestimate, and explains how to perform reasonable gamma dose calculations averaged over 10 cm2. The article also describes upgrades underway in Varskin 3. PMID- 14744064 TI - Glow curve evaluation in routine personal dosimetry. AB - The glow curve of LiF:Mg, Ti (TLD-100) has a well known and defined shape. Several factors, however, can distort the glow curve shape and can lead to incorrect dosimetry. Only proper evaluation of glow curves, either by visual inspection or computerized method, can detect these anomalies. Visual observation is riot only tedious and time consuming but may also be susceptible to human errors. The usefulness of a commercially available glow curve analysis software during quality control testing of thermoluminescence dosimetry cards is discussed. The long-term stability of the thermoluminescence dosimetry reader is also investigated using this software. Various types of glow curve anomalies encountered during routine personal dosimetry practice are presented. PMID- 14744065 TI - Radioactivity in cathode ray tubes. AB - While surveying used computer equipment out of a zone posted as a Contamination Area, 100% of the computer monitors surveyed had levels of radioactivity that were significantly above background. The radioactivity was primarily on the front face of the cathode ray tube and was not amenable to decontamination. Hot spots were found also along the edges and seals of the cathode ray tube. Similar surveys of computer monitors that were never in Contamination Areas confirmed that radioactivity was incorporated into the monitor. Surveys were made of recently manufactured television sets with similar results. Gamma spectroscopy indicates that the radioactivity is due to naturally occurring radioactive materials. Since most surveys of cathode ray tubes in the literature were made while the units were energized and indicated low-energy x-rays, the use of naturally occurring radioactive materials in the manufacture of cathode ray tubes has not been widely recognized. This paper presents the results of these surveys, the results of gamma spectroscopy, and a method for releasing existing computer equipment having naturally occurring radioactive materials. PMID- 14744066 TI - Summing coincidence errors using 152Eu lungs to calibrate a lung-counting system: are they significant? AB - The use of a lung phantom containing 152Eu/241 Am activity can provide a sufficient number of energy lines to generate an efficiency calibration for the in vivo measurements of radioactive materials in the lungs. However, due to the number of energy lines associated with 152Eu, coincidence summing occurs and can present a problem when using such a phantom for calibrating lung-counting systems. A Summing Peak Effect Study was conducted at three laboratories to determine the effect of using an efficiency calibration based on a 152Eu/241 Am lung phantom. The measurement data at all three laboratories showed the presence of sum peaks. While one of the laboratories found only small biases (< 5%) when using the 152Eu/241 Am calibration, the other facilities noted up to 30% positive bias in the 140 keV to 190 keV energy range that prevents the use of the 152Eu/241 Am lung phantom for routine calibrations. Although manufactured by different vendors, the three facilities use similar types of germanium detectors (38 cm2 by 25 mm thick or 38 cm2 by 30 mm thick) for counting. These results underscore the need to evaluate the coincidence summing effect, which appear system dependent, when using a nuclide such as 152Eu for the calibration of low energy lung counting systems and highlight the problem of using a general calibration curve in place of specific nuclide calibration factors. PMID- 14744067 TI - Sentinel node biopsy: ALARA and other considerations. AB - For a majority of solid tumors, the most powerful and predictive prognostic factor is the status of the regional lymph nodes. Sentinel lymph node sampling continues to gain in popularity as patients and their physicians seek to avoid the potential morbidity associated with standard axillary node dissection. Lymphoscintigraphy, one of the recently explored techniques of lymphatic mapping, involves pre-operative intradermal or subcutaneous administration of a radiopharmaceutical. While this approach is gaining widely spread acceptance, there is still a lack of consensus on which radiopharmaceutical agent has the most ideal properties. By far, the most commonly used agents are 99mTc labeled colloids, but other agents are also used clinically and are under investigation or development worldwide. A number of other clinical, technical, dosimetric, and logistical considerations regarding this procedure remain. They include questions such as who should be performing the procedure, what precautions to take during surgery, how to better isolate "hot" nodes and thus improve the efficacy of determining metastases to the draining lymph node, what precautions to take when handling surgical specimens, etc. There is clearly a need to review as low as reasonably achievable considerations and other issues that arise as this technique evolves and finds its role in the evaluation of various types of cancers. This paper, based on our own experiences and those of others, fills this gap. PMID- 14744068 TI - Radiation safety precautions for sentinel lymph node procedures. AB - Radiation safety professionals need guidelines for controlling occupational radiation doses and handling generated contaminated tissues from sentinel lymph node procedures involving 99mTc. Many healthcare facilities do not have formal controls in place, or they adopt policies that may be too restrictive or impractical. During accreditation inspections at healthcare facilities, agencies such as the College of American Pathology and the Joint Commission on the Accreditation of Healthcare Organizations occasionally raise questions about these issues. Inspectors expect to find formal policies to ensure these procedures are performed safely. The objective of this article is to review the basic radiation safety controls needed during sentinel lymph node procedures and to propose guidelines that may be used to control occupational radiation doses and handling of SLN contaminated tissues. PMID- 14744069 TI - An unusual incident: breached 22Na sealed source. AB - A typical laboratory contamination with radioactive materials in an academic institution is the result of a spill or other handling errors with unsealed radioactive materials. An unusual incident occurred in late January 2002 at our institution when a 22Na sealed source was breached, and a small portion of the contents accidentally migrated to various locations in the very large laboratory. Damage control provided a challenge to radiation safety personnel. Resources of the Radiation Safety Office were severely taxed during both the immediate reaction and the subsequent several months of decontamination prior to release of the laboratory for unrestricted use of radioactive materials once again. Salient features of this incident are described in conjunction with a portrayal of measures taken during early damage control and the following deliberate remediation. PMID- 14744070 TI - Radiation risks and dirty bombs. AB - For many, the thought of terrorists detonating a dirty bomb--a radiological dispersal device--is frightening. However, the radiation health risks from such an occurrence are small. For most people directly involved, the exposure would have an estimated lifetime health risk that is comparable to the health risk from smoking five packages of cigarettes or the accident risk from taking a hike. The actual impact of a dirty bomb would be economic and social (NCRP 2001). There would be an economic cost for clean-up as well as a decrease in economic activity in the affected area due to radiation fear. If such a bomb were detonated, those exposed as well as those not exposed would have great concern about potential health effects while seeking medical attention and avoiding the impacted area. This paper discusses the health risks from radiation exposure and compares them to risks from various activities of daily life and to exposure to hazardous chemicals. PMID- 14744071 TI - Legal liability and the health physicist. PMID- 14744072 TI - Routine dose estimates for the removal of soil from a basin to the burial ground at the Savannah River Site. AB - Worker dose estimates have been made for various exposure scenarios resulting from the relocation of soil from the H Area Retention Basin to the Old Radioactive Waste Burial Ground at the Savannah River Site. Estimates were performed by hand calculations and using RESRAD and MAXDOSE-SR. Doses were estimated for the following pathways: (1) shine and inhalation as a result of standing on contaminated soil at the H Area Retention Basin and the Old Radioactive Waste Burial Ground; (2) exposure to off-unit receptors due to soil disturbances from excavation type activities at the H Area Retention Basin and the Old Radioactive Waste Burial Ground; (3) exposure to off-unit receptors due to soil disturbances from dumping of soil from bucket and from roll-off pan; and (4) exposure to off-unit receptors from wind driven dust from contaminated area. The highest dose estimates (0.25 mSv h(-1)) resulted from the receptor standing on the H Area Retention Basin. PMID- 14744073 TI - The voluntary siting process, a case study in New Jersey. AB - The Low-Level Radioactive Waste Policy Act of 1980 and its 1985 Amendments has not provided new disposal capacity within the United States; however, sufficient disposal capacity currently exists to handle today's disposal needs. Politics, opposition groups, and public mistrust in government have combined to limit the possibilities for establishing new disposal facilities. In 2000, New Jersey (NJ and Connecticut (CT), as members of the Northeast Compact for the disposal of low level radioactive waste, admitted South Carolina (SC) to their compact, renaming it as the "Atlantic Compact." The advantage to SC is that they are able to prevent disposal of waste from outside the Compact. The advantage to NJ and CT is that they are guaranteed waste disposal for approximately the next 50 years, or until all currently operating nuclear power plants in the states are decommissioned. This paper details the process, much of it not following the scientific method, to try to site a low-level waste facility in NJ. With the formation of the NJ Siting Board in 1987, an effort was made to locate a site using deterministic criteria; however, in 1992, the Board shifted to a voluntary process. In 1998, the Board made the determination that there was adequate capacity for waste disposal and ended active siting. In 2000, the opportunity to form the Atlantic Compact ended siting through an out-of-state solution. While it is not clear that the voluntary process would have ultimately worked in NJ, it has worked in Canada and the process may be one of the few mechanisms for the siting of any type of hazardous material disposal facility. Also, other states still have to decide what they will do after 2008 when Barnwell is no longer open to them. PMID- 14744074 TI - Personality disorders over time: precursors, course and outcome. AB - Personality disorders cause dysfunction over the course of adult life. A chronic course of disorder tends to be associated with an early onset, and personality disorders are preceded by precursor symptoms in childhood. Long-term outcome varies by personality disorder category: antisocial and borderline personality tend to remit with age, an improvement that is not seen in other diagnoses. The chronicity of personality disorders can usefully guide treatment planning, and psychotherapy for personality disorders can focus on rehabilitation. PMID- 14744075 TI - Dangerous and severe personality disorders: a new personality concept from the United Kingdom. AB - In 1998, we in England were shocked by the apparently motiveless murders of a mother and two of her children when they were traveling home from school in a rural area. Subsequently a patient with personality disorder, being monitored by the forensic psychiatric services, was convicted of their murders. The government was determined to prevent this type of offense from recurring and in 1999 introduced a new concept, dangerous and severe personality disorder (DSPD). This subsequently became a treatment and assessment program for individuals who satisfy three requirements: (1) have a severe disorder of personality, (2) present a significant risk of causing serious physical or psychological harm from which the victim would find it difficult or impossible to recover, and (3) the risk of offending should be functionally linked to the personality disorder. The implications of this program are discussed with regard to international systems of classification and service provision. We also attempt to place DSPD in the wider context of psychiatric participation in the management of personality disordered offenders. PMID- 14744076 TI - Frontal lobe dysfunctions in borderline personality disorder? Neuropsychological findings. AB - This study aims to determine whether specific neuropsychological performance impairments in borderline patients can be objectified and whether these findings indicate frontal dysfunctions. Twenty-three patients with borderline personality disorder and 23 normal controls were examined using a neuropsychological test battery to assess intelligence, attentiveness, proneness to interference, learning and memory, as well as planning and problem solving. All subjects filled out standardized questionnaires to assess aggressiveness and impulsiveness in the context of these cognitive performance areas. The neuropsychological test results of the borderline patients were comparable to those of the controls. Although there were no indications of frontal dysfunction of cognitive information processing, inverse correlations were found between the severity of borderline related personality traits regarding impulsiveness and various areas of cognitive performance. Borderline personality patients show no indications of frontal cognitive dysfunction. Further research is needed to clarify the relationship between impulsiveness and cognitive information processing in borderline personality disorder, including a dimensional approach to personality and personality disorder. PMID- 14744077 TI - Outpatient group psychotherapy following day treatment for patients with personality disorders. AB - This prospective, naturalistic study evaluated the practice and effectiveness of an outpatient group therapy program following day treatment for patients with personality disorders (PDs). One hundred and eighty-seven patients (86% patients with PDs and 14% with no PDs), were treated in outpatient psychodynamic group therapy. Outcome was assessed by Global Assessment of Functioning, Symptom Check List 90-R, and Inventory of Interpersonal Problems-Circumplex, short version, at admission and discharge from day treatment, and at the end of outpatient group therapy. Average length of outpatient therapy was 24 months. Forty-three percent terminated in an irregular manner. Outcome of the continuation therapy was satisfactory for patients without PDs. For PD patients, the improvement from the day treatment was maintained during outpatient therapy, but further improvements were modest for symptoms and interpersonal distress, somewhat better for global functioning. Implications for further treatment development are discussed. PMID- 14744078 TI - Parental bonding and personality pathology assessed by clinician report. AB - The present study focused on the link between parental bonding and personality pathology. We developed a clinician-report version of the Parental Bonding Instrument (PBI; Parker, Tupling, and Brown, 1979), which 203 clinicians applied to a randomly selected patient in their care. The goal of the study was to examine the reliability and validity of a clinician-report PBI, and to provide a preliminary examination of the relation between personality disorder (PD) symptoms and parental bonding in a national sample of patients in treatment in the community. Factor analysis yielded two factors (parental rejection/care and parental overprotection) for each parent, which had high internal consistency (coefficient alphas all >.80). The factor structure of the clinician-report measure strongly resembled the factor structure of the widely used self-report instrument. Parental bonding as assessed by clinician report showed coherent patterns of association with PD diagnoses, which resembled those found in other samples. Maternal rejection/care showed a particular link to borderline PD (BPD), and remained a significant predictor of BPD along with sexual abuse and physical abuse in multiple regression analysis. These data, along with others from recent studies using clinicians as informants, suggest that clinicians can provide reliable and valid data when their observations are quantified using psychometrically sound instruments. Clinician-report methods may provide a useful complement to studies of personality disorders that rely primarily on self reports or structured interviews. PMID- 14744079 TI - Personality factors and posttraumatic stress: associations in civilians one year after air attacks. AB - There is an ongoing debate on which risk factors for developing posttraumatic stress symptoms are more important--personality traits reflecting vulnerability, previous stressful experiences or characteristics of the traumatic event. In this study, posttraumatic stress symptoms and their relationship with personality traits, previous stressful experiences and exposure to stressful events during air attacks in Yugoslavia were investigated. The Millon Clinical Multiaxial Inventory (MCMI; Millon, 1983), Impact of Events Scale (IES; Horowitz, Wilner, & Alvarez, 1979), Life Stressor Checklist Revised (LSCL-R; Wolfe & Kimerling, 1997), and List of Stressors were administered to a homogeneous group of medical students 1 year after the attacks. In multiple regression analyses, compulsive and passive-aggressive personality traits and a higher level of exposure to stressors during air attacks independently predicted the degree of intrusion symptoms. Avoidance symptoms were predicted by avoidant personality traits and a higher exposure to stressors both previously in life and during the attacks. In the next step, we tested in analyses of variance whether personality traits, previous stressful experiences, and stressful events during attacks as independent variables interact in predicting intrusion and avoidance symptoms. For this, students were clustered into three groups depending on their predominant personality traits. In addition to direct predictive effects, there were significant interaction effects in predicting both intrusion and avoidance. The findings suggest that each of the tested factors, i.e., personality traits, previous stressful experiences, and exposure to traumatic events may have an independent and direct influence on developing posttraumatic stress. However, the effect of these factors cannot just be added up. Rather, the factors interact in their impact on posttraumatic stress symptoms. Bigger samples and longitudinal designs will be required to understand precisely how different personality traits influence response to stressful events. PMID- 14744080 TI - Assessment of DSM-IV personality disorders in obsessive-compulsive disorder: comparison of clinical diagnosis, self-report questionnaire, and semi-structured interview. AB - In patients with obsessive-compulsive disorder, personality disorders are not many times assessed according to DSM-IV criteria. The purpose of the present study is to examine the prevalence of personality disorders diagnosed according to the DSM-IV in a severely disordered OCD population (n=65) with three different methods of assessing personality disorders (structured interview, questionnaire, and clinical diagnoses). Furthermore, correspondence between these different methods was investigated and their construct validity was examined by relating the three methods to external variables. Each method resulted in a predominance of Cluster C personality disorders, and obsessive-compulsive personality disorder had the highest prevalence. However, there was generally low correspondence regarding which patient had which personality disorder. Results concerning the relation of external variables were the most promising for the structured clinical interview. PMID- 14744081 TI - Short-interval test-retest interrater reliability of the Dutch version of the Structured Clinical Interview for DSM-IV personality disorders (SCID-II). AB - This study examined the short-interval test-retest reliability of the Structured Clinical Interview (SCID-II: First, Spitzer, Gibbon, & Williams, 1995) for DSM-IV personality disorders (PDs). The SCID-II was administered to 69 in- and outpatients on two occasions separated by 1 to 6 weeks. The interviews were conducted at three sites by ten raters. Each rater acted as first and as second rater and equal number of times. The test-retest interrater reliability for the presence or absence of any PD was fair to good (kappa = .63) and was higher than values found in previous short-interval test-retest studies with the SCID-II for DSM-III-R. Test-retest reliability coefficients for trait and sumscores were sufficient, except for dependent PD. Values for single criteria were variable, ranging from poor to good agreement. Further large-scale test-retest research is needed to test the interrater reliability of more categorical diagnoses and single traits. PMID- 14744082 TI - A screening measure for BPD: the McLean Screening Instrument for Borderline Personality Disorder (MSI-BPD). AB - Borderline personality disorder (BPD) is a common psychiatric disorder that is often overlooked in treatment settings. This report describes the validation of a new self-report screening measure for DSM-IV BPD--the McLean Screening Instrument for Borderline Personality Disorder (MSI-BPD). Two hundred subjects with treatment histories whose ages ranged from 18 to 59 filled out the MSI-BPD. Each subject was then interviewed, blind to MSI-BPD results, with the BPD module of the Diagnostic Interview for DSM-IV Personality Disorders (DIPD-IV; Zanarini, Frankenburg, Sickel, & Yong, 1996). Of these 200 subjects, 139 (69.5%) met DSM-IV criteria for BPD as assessed by the BPD module of the DIPD-IV and the remaining 61 subjects (30.5%) did not. Using logistic regression analyses, an MSI-BPD cutoff of 7 or more of the measure's 10 items was judged to be the best cutoff. This was so because it yielded both good sensitivity (.81; percentage of correctly identified cases) and specificity (.85; percentage of correctly identified noncases) for the diagnosis of DSM-IV BPD. For younger subjects, diagnostic efficiency was even greater. For example, sensitivity was .90 and specificity was .93 at a cutoff of 7 for the 63 subjects who were 25 years old or younger. The results of this study suggest that the MSI-BPD may be a useful screening instrument for the presence of DSM-IV borderline personality disorder. PMID- 14744084 TI - Histologic features of melanocytic nevi seen in association with mycosis fungoides. AB - BACKGROUND: Many different tumors have been reported to occur simultaneously as collision lesions. To date, no such events have been reported between mycosis fungoides (MFs) and melanocytic neoplasms. METHODS: Two cases are presented in which patches of MF were superimposed on melanocytic nevi. In addition, 967 biopsies of MF from 411 patients were identified in an 8-year retrospective database search. Patient pathology history summaries were reviewed to identify inflamed nevi, atypical nevi, and melanoma submitted for histologic evaluation from this population. RESULTS: The occurrence of MF in a congenital nevus was associated with a halo phenomenon restricted to the affected region of the nevus in one patient. In the other patient, nests of two morphologies (lymphocytic and melanocytic) in the same biopsy presented a potentially confusing histologic picture. No other cases of MF superimposed on a nevus were identified in 967 biopsies from 411 patients with a histological diagnosis of MF seen over the past 8 years. In this population, 57 biopsies of melanocytic lesions were identified from 28 patients, including three atypical nevi and three melanomas. CONCLUSIONS: The presence of MF superimposed on a nevus is rare and may lead to confounding histologic features or the development of a halo nevus phenomenon. PMID- 14744083 TI - Expression of insulin-like growth factor-binding protein 2 in melanocytic lesions. AB - BACKGROUND: Insulin-like growth factor-1 (IGF-1) is one of the most critical proteins required for the survival, migration, and growth of melanoma cells. IGF binding protein 2 (IGFBP2), which binds and regulates the function of IGF-1, is upregulated in a dose-dependent manner in melanoma cells treated with IGF-1, suggesting a possible role of IGFBP2 in the pathogenesis of melanoma. METHODS: Tissue microarrays were constructed using formalin-fixed, paraffin-embedded archival tissue blocks from 94 melanocytic lesions: 20 benign nevi, 20 dysplastic nevi, 23 primary melanomas, and 31 metastatic melanomas. IGFBP2 expression was evaluated immunohistochemically using a polyclonal antibody against the C terminus of IGFBP2. The number of cells and labeling intensity were assessed semiquantitatively. RESULTS: Positive IGFBP2 labeling was observed in 5.0% of benign nevi, which was significantly lower than in dysplastic nevi (35.0%), primary melanomas (52.2%), or metastatic melanomas (54.8%) (p < 0.05). Among the IGFBP2-positive cases, moderate-to-strong immunostaining was observed in 64.7% of metastatic melanomas and 33.3% of primary melanomas. But none of the dysplastic nevi had moderate-to-strong immunostaining (p < 0.05). CONCLUSIONS: Our study shows that IGFBP2 expression increases from benign and dysplastic nevi to primary and metastatic melanomas and suggests that it may play a role in melanoma progression. PMID- 14744085 TI - The periodic acid-Schiff stain in diagnosing tinea: should it be used routinely in inflammatory skin diseases? AB - BACKGROUND: The clinical presentation of tinea is usually, but not always, characteristic. The purpose of this study was to determine whether the routine use of the periodic acid-Schiff (PAS) stain in inflammatory skin disorders is requisite for identifying clinically undiagnosed tinea cases and to ascertain whether there are histopathologic clues that suggest the diagnosis of a dermatophyte infection. METHODS: Hematoxylin and eosin (H&E)-stained slides from 60 PAS-positive tinea cases were examined histologically by two observers. One observer, aware of the diagnosis of tinea, searched for hyphal elements and also recorded in detail epidermal, dermal, and follicular changes. The second observer, not aware of the diagnosis beforehand, reviewed the same slides, together with randomly mixed slides from 21 non-tinea cases, recording the same parameters as the first reviewer. RESULTS: Of the 60 cases of tinea, only 45% were diagnosed clinically. Histologic examination of H&E sections by the two observers disclosed the presence of hyphal elements in 68 and 45%, respectively. No significant histologic differences, except for the presence of hyphae, were observed between tinea and non-tinea cases. CONCLUSIONS: The finding that only 57% of PAS-positive cases of tinea showed hyphal elements on H&E examination alone, together with no other differentiable histologic characteristics, lends strong support for the routine use of PAS-staining for inflammatory skin disorders. PMID- 14744086 TI - Identification of parasite antigen, correlation of parasite density and inflammation in skin lesions of post kala-azar dermal leishmaniasis. AB - BACKGROUND: Post kala-azar dermal leishmaniasis (PKDL) is an unusual dermatosis following kala-azar (KA). Demonstration of the amastigotes in lesions plays an important role in the diagnosis of PKDL. It was aimed to evaluate the utility of an antibody G2D10 in detecting leishmania parasite antigen, to correlate the parasite number/percentage of parasites with the inflammation, and to assess the epidemiological significance associated with the location of the parasites. MATERIAL AND METHODS: The study was conducted on 50 cases. Hematoxylin and eosin (H & E) stains and immunohistochemical (IHC) stains, using G2D10 antibody, was performed on the skin biopsies. The number of parasites and density of inflammation were semiquantitatively assessed. RESULTS: Leishmania donovan bodies (LDBs) were identified in 50% of cases with the H & E compared to 80% positivity with the IHC. All 50 cases showed inflammation in the superficial dermis (SD). About 44% showed dense inflammation compared to 16% sparse and 40% moderate inflammation. Parasite percentage was maximum in the SD (100%) compared to 75 and 42% in the mid and deep dermis, respectively. CONCLUSIONS: The IHC showed a higher percentage of LDB localization (80 vs. 50%). Density of inflammation was maximum in the SD. The parasite percentage was correlated with the inflammation. Location of parasites could have an epidemiological significance. PMID- 14744087 TI - Expression of desmosomal proteins in squamous cell carcinomas of the skin. AB - BACKGROUND: Desmosomal proteins are well established markers of epithelial differentiation. Down-regulation of desmosomal proteins has been suggested to be a sign of reduced adhesiveness in metastasizing cells. METHODS: We examined actinic keratoses, Bowen's disease, and squamous cell carcinoma (SCC) of the skin for the expression of desmosomal proteins using isoform-specific antibodies on paraffin-embedded sections. Evaluation was performed qualitatively in comparison to the epidermis and semiquantitatively using an area-intensity-score. RESULTS: We found no qualitative correlation of desmoplakin or plakoglobin expression with risk of metastasis. Plakophilin 1, desmoglein 1, and the desmocollins 1-3 were found to be heterogeneously expressed in all neoplasms without significant correlation to aggressive tumor behavior. Plakophilin 2 was not expressed in any of the neoplasms examined. As most striking finding, desmoglein 2 was up regulated qualitatively in half of all neoplasms examined and showed a significant higher proportion of positive cells in high-risk SCC than in low-risk SCC. CONCLUSIONS: Desmosomal proteins are highly regulated in cutaneous SCC. Only desmoglein 2 expression correlates with risk of metastasis. Desmosomes may still be functional in metastasizing tumor cells. PMID- 14744088 TI - Solitary sclerotic fibroma of skin: a possible link with pleomorphic fibroma with immunophenotypic expression for O13 (CD99) and CD34. AB - BACKGROUND: Solitary sclerotic fibroma (SF) presents as a well circumscribed dermal nodule, composed of sparse spindle cells with alternating wavy collagen fibers arranged in a storiform pattern. The histogenesis and nature of this histologically distinct lesion are uncertain. Whether this peculiar tumor represents a true hamartoma or a degenerating end of various fibrous lesions such as pleomorphic fibroma (PF), dermatofibroma, or angiofibroma is still controversial. High proliferating index of spindle cells in SF argues against the possibility of being a degenerating end product of another lesion. METHODS: We studied morphological features and immunoprofile of eight SFs, in comparison with four PFs, one collagenized dermatofibroma, two angiofibromas, and two periungual fibromas. Immunostains for CD34, CD31, O13 (CD99), Factor XIIIa, S-100, CD68 (KP 1), and MIB-1 were carried out using a labeled streptavidin-biotin method with DAKO-automated immunostainer. Paraffin blocks of two SFs were reprocessed for electron microscopic studies. Clinical data of all patients with SF were also reviewed. RESULTS: Spindle cells and pleomorphic cells in SF and PF showed diffuse immunoreactivity for CD34 and O13 but were negative for CD31, S-100, and CD68. Spindle cells in one dermatofibroma and one angiofibroma were positive for Factor XIIIa. Proliferating index (MIB-1) was very low in all cases of SF, contradicting some previous reports. CONCLUSIONS: SF is a fibrotic lesion with cells positive for CD34 and O13. It shares a common immunoprofile with PF but is distinct from dermatofibroma and other common spindle cell lesions of skin. O13 expression in SF has not been previously described. PMID- 14744089 TI - Cutaneous metastasis of hepatocellular carcinoma diagnosed with hepatocyte paraffin (Hep Par 1) antibody immunohistochemistry. AB - Cutaneous metastases from hepatocellular carcinomas are rare, and their diagnosis may be difficult on histological grounds. We report a case of metastatic hepatocellular carcinoma to the skin that was confirmed immunohistochemically by the expression of a hepatomitochondria-specific antigen detectable on paraffin embedded sections (Hep Par 1). PMID- 14744091 TI - Metastatic nevoid melanoma in a 4 1/2-year-old child. AB - BACKGROUND: Childhood melanoma is a rare and controversial diagnosis. METHODS: We present the case of a 4 1/2-year-old child found to have an expanding, elevated pigmented lesion on her back. RESULTS: The biopsy showed a symmetrical, well circumscribed lesion. However, higher magnification revealed sheets of nevoid cells infiltrating deep into the dermis, lacking maturation, and exhibiting a high mitotic rate (average 5/10 high-power field) with deep mitoses. The possibility of nevoid melanoma was raised, and re-excision and sentinel lymph node (SLN) biopsy were recommended. Two SLNs were positive for melanoma, verified by immunohistochemical staining. In order to further characterize this melanoma, we performed immunohistochemistry for the tumor-suppressor p53, proliferation marker MIB-1, and oncogenes Bcl-2, cyclin D-1, and MDM-2. Staining for p53 was diffusely positive in the primary and the metastasis; MIB-1 showed moderate proliferative rate in the primary (approximately 10%); Bcl-2 was weakly positive in the primary and showed focal staining in the metastasis; cyclin D-1 was strongly positive in the primary and metastasis; and MDM-2 staining showed scattered positive cells in both lesions. CONCLUSIONS: These findings are consistent with a metastatic nevoid melanoma arising in the absence of predisposing disease in a young child, a distinctly unusual occurrence. PMID- 14744090 TI - Epidermotropic metastases from breast carcinoma showing different clinical and histopathological features on the trunk and on the scalp in a single patient. AB - A 54-year-old female presented with the cutaneous metastases of the breast carcinoma that produced combination of pigmented zosteriform eruption on the trunk and eroded plaque on the scalp, 13 years after radical mastectomy. Histologically, zosteriform lesions displayed prominent infiltration of the epidermis in nesting or linear pattern by neoplastic cells with focal formation of intraepidermal and subepidermal vesicles due to discohesion of tumor cells and dermal edema. Examination of scalp plaque revealed ulcerations and infiltration of the epidermis with scattered basal and suprabasal malignant cells in pagetoid fashion. Immunohistochemically, tumor cells were cytokeratin 7- and estrogen receptor-positive and cytokeratin 20 negative. HMB-45 and Melan-A-stained numerous dendritic melanocytes intermingled with intraepidermal and superficial dermal tumor cells in the trunk lesion, whereas on the scalp, only occasional melanocytes surrounding intraepidermal carcinomatous cells were identified. Our case described, to our knowledge, so far unreported combination of individually rare, clinical and histological patterns of cutaneous metastases from breast carcinoma in a single patient. PMID- 14744092 TI - Mucoepidermoid carcinoma arising within nevus sebaceus of Jadassohn. AB - BACKGROUND: Nevus sebaceus (NS) of Jadassohn is a common congenital lesion associated with numerous benign and malignant tumors. However, mucoepidermoid carcinoma (MEC) has not been described in association with NS. METHODS: We describe an unusual case of MEC arising within NS. RESULTS: A 72-year-old man presented with an enlarging plaque on his forehead, along the hairline. Physical examination revealed a mounded, erythematous lesion that was completely excised. Histological evaluation revealed a typical MEC with areas of squamous and adenomatous differentiation with foci of typical intermediate cells. CONCLUSION: The diagnosis of MEC is an important one, because it portends a poor prognosis, requiring long follow-up. This is, to our knowledge, the first report of MEC arising within NS. PMID- 14744093 TI - Foreword: from the TRANSCRIPTOME conferences to the SYSTEMOSCOPE international consortium. AB - This thematic issue issue of the Comptes rendus Biologies contains review articles, original papers and conference reports presented at the first two TRANSCRIPTOME conferences From Functional Genomics to Systems Biology and IMAGE Consortium Invitational workshops (Paris, November 2000 and Seattle, March 2002), and discussed during the inaugural meetings of the SYSTEMOSCOPE International Consortium (Paris, June 2003). We describe the founding principles, missions, working plan and policy for partnership and industrial development of SYSTEMOSCOPE to promote the study of the complexity of biological systems by integrating scientific, medical, ethical and economic issues in implementation of interdisciplinary projects for human health. PMID- 14744094 TI - From functional genomics to systems biology: concepts and practices. AB - Systems biology is the iterative and integrative study of biological systems as systems in response to perturbations. It is founded on hypotheses formalized in models built from the results of global functional genomics analyses of the complexity of the genome, transcriptome, proteome, metabolome, etc. Its implementation by cross-disciplinary teams in a standardized mode under quality assurance should allow accessing the small variations of the large number of elements determining functioning of biological systems. Galactose utilization in yeast, and sea urchin development are two examples of emerging systems biology. PMID- 14744095 TI - From the messenger RNA saga to the transcriptome era. AB - This review attempts to provide an overview of the evolution of the ideas and techniques that prevailed at the beginning of research on ribonucleic acids, until the contemporary era of cellular transcript analysis using DNA biochips and microarrays. Certain applications derived from the use of microarrays and the corresponding analyses of transcriptomes are discussed, particularly concerning diagnosis and prevision of evolution of certain cancers. PMID- 14744096 TI - Expanding genome capacity via RNA editing. AB - RNA editing, which results in the creation of RNA molecules that differ from the template from which they were made, is a highly specific process. Alterations include converting one base to another, removal of one nucleotide and substitution of another, deletion of encoded residues, and insertion of non templated nucleotides. Such changes have marked effects on gene expression, ranging from defined amino acid changes to the de novo creation of entire open reading frames. Editing can be regulated in a developmental or tissue-specific manner, and is likely to play a role in the etiology of human disease. PMID- 14744097 TI - Transcriptomic analysis of the NCI-60 cancer cell lines. AB - Pharmacogenomics aims at molecular subsetting of patients for more effective therapy. Transcriptomic profiling of the 60 human cancer cell lines (the NCI-60) used by the US National Cancer Institute serves that aim because the cells have been treated with > 100,000 chemical compounds over the last 13 years. Patterns of potency can be mapped into molecular structures of the compounds or into molecular characteristics of the cells. We discuss conceptual and experimental aspects of the profiling, as well as a number of bioinformatic computer programs that we have developed for biological interpretation of the profiles. PMID- 14744098 TI - The Riken mouse genome encyclopedia project. AB - The Riken mouse genome encyclopedia a comprehensive full-length cDNA collection and sequence database. High-level functional annotation is based on sequence homology search, expression profiling, mapping and protein-protein interactions. More than 1000000 clones prepared from 163 tissues were end-sequenced and classified into 128000 clusters, and 60000 representative clones were fully sequenced representing 24000 clear protein-encoding genes. The application of the mouse genome database for positional cloning and gene network regulation analysis is reported. PMID- 14744099 TI - The NIA cDNA project in mouse stem cells and early embryos. AB - A catalog of mouse genes expressed in early embryos, embryonic and adult stem cells was assembled, including 250000 ESTs, representing approximately 39000 unique transcripts. The cDNA libraries, enriched in full-length clones, were condensed into the NIA 15 and 7.4K clone sets, freely distributed to the research community, providing a standard platform for expression studies using microarrays. They are essential tools for studying mammalian development and stem cell biology, and to provide hints about the differential nature of embryonic and adult stem cells. PMID- 14744100 TI - Transcriptome study in China. AB - The Chinese genome project was initiated in 1993 with the goal of contributing 1% to the Human Genome Program. The study of gene expression profiles with cDNA microarrays, and large-scale sequencing and analysis of 130928 expressed sequence tags (ESTs), allowed isolation and characterization of over 1000 novel full length human cDNAs derived from human hematopoietic stem/progenitor cells, neuroendocrine tissues, liver, and cardiovascular cells. In addition, EST sequencing for model organisms, including rat, zebrafish, Schistosoma japonicum and rice was performed, aiming at identifying genes associated with physiological and/or pathological characteristics. PMID- 14744101 TI - The Kazusa cDNA project for identification of unknown human transcripts. AB - The Kazusa cDNA project is unique by its focus on sequencing large human cDNAs (>4 kb). We describe an overview of the human cDNA sequence data accumulated during the first phase of the project on over 2000 cDNAs and its integration with the genome sequence. In the second phase of the project, which aims at bridging the human genome and proteome using the output of the first phase, we are very carefully evaluating our cDNA clones and, when necessary, experimentally revising them. PMID- 14744102 TI - LNCIB human full-length cDNAs collection: towards a better comprehension of the human transcriptome. AB - LNCIB has been producing a variety of human full-length-enriched, normalized and subtracted cDNA libraries from various cell lines and tissues in different developmental stages by using the CAP-Trapper method. By sequencing 23000 clones of these libraries we identified a pool of about 5800 good quality unique cDNAs. After BLAST analysis on Human RefSeq/Unigene databases, 1717 of these sequences remained with no or poor annotation. We show that cross-species comparative BLAST resulted as a valid tool for the annotation of orthologous genes. PMID- 14744103 TI - Concatenation cDNA sequencing for transcriptome analysis. AB - We describe a high-throughput cDNA sequencing pipeline (http://www.hgsc.bcm.tmc.edu/projects/cdna) built in response to the emerging need for rapid sequencing of large cDNA collections. Using this strategy cDNA inserts are purified and joined through concatenation into large molecules. These 'pseudo-BACs' are subjected to random shotgun sequencing whereby the majority of cDNA inserts in the pool are sequenced. Using this concatenation cDNA sequencing platform, we have contributed more than 13000 full-length cDNA sequences from human and mouse to the Mammalian Gene Collection (MGC). PMID- 14744104 TI - ORESTES are enriched in rare exon usage variants affecting the encoded proteins. AB - A significant fraction of the variability found in the human transcriptome is due to alternative splicing, including alternative exon usage (AEU), intron retention and use of cryptic splice sites. We present a comparison of a large-scale analysis of AEU in the human transcriptome through genome mapping of Open Reading Frame ESTs (ORESTES) and conventional ESTs. It is shown here that ORESTES probe low abundant messages more efficiently. In addition, most of the variants detected by ORESTES affect the structure of the corresponding proteins. PMID- 14744105 TI - Small open reading frames in 5' untranslated regions of mRnas. AB - Using the 5'-end sequence data from 'oligo-capped' cDNAs, we generated a representative full-length cDNA dataset for 4870 RefSeq entries, and analyzed the 5' untranslated region (UTR) of these genes. To our surprise, about half of the 4870 genes had an upstream ATG before the ATG that starts the longest open reading frame (ORF), suggesting that about half of them have small ORFs in their 5' UTR of average length of 31 amino acids. They require attention for further analysis to identify their biological role. PMID- 14744106 TI - High-throughput classification of images of cells transfected with cDNA clones. AB - The sequence of the human genome has been determined. The next task is to determine the function of the genes. Classifying cellular forms of proteins encoded by human cDNA clones is a primary step toward understanding the biological role of proteins and their coding genes. We report here our ongoing work on an automatic system to facilitate this classification. Our system handles the transfection, incubation, acquisition of microscopic images of the cells, and the classification of forms there appearing in the images. Our system correctly classified proteins by their forms at a rate of 90% in feasibility studies. PMID- 14744107 TI - CDNAs for functional genomics and proteomics: the German Consortium. AB - To functionally characterize numerous novel proteins encoded by cDNAs sequenced by the German Consortium, 800 were tagged with green fluorescent protein. The subcellular localizations of the fusion proteins were examined in living cells, enabling their classification in subcellular groups. Their activity in cell growth, cell death, and protein transport was screened in high throughput using robotic liquid handling and reading stations. The resulting information is integrated with functional genomics and proteomics data for further understanding of protein functions in the cellular context. PMID- 14744108 TI - In situ analysis of gene expression in Xenopus embryos. AB - The molecular anatomy of the vertebrate embryo was systematically analysed through gene expression during early development of the Xenopus frog using whole mount in situ hybridization. Expression patterns are documented and assembled into the database Axeldb (http://www.dkfz-heidelberg.de/abt0135/axeldb.htm). Synexpression groups representing genes with shared, complex expression pattern that predict molecular pathways involved in patterning and differentiation have been identified. These sets of co-regulated genes show a striking similarity with operons, and may be a key determinant facilitating evolutionary change leading to animal diversity. PMID- 14744109 TI - RNA amplification, fidelity and reproducibility of expression profiling. AB - We quantitatively address the effect of T7 RNA amplification on expression profiling data, and answer the following questions: (1) What fraction of genes sampled is amplified non-linearly? (2) What is the effect of RNA amplification on comparative expression measurements? (3) If there is amplification bias, is the bias dependent on the degree of amplification or the amount of starting material and (4) Does amplification increase the overall variability of the results? We show that while there is significant amplification bias, the bias is consistent and generally has little effect on array comparisons between amplified samples. PMID- 14744110 TI - Gene expression profiling of breast carcinomas using nylon DNA arrays. AB - Clinically very heterogeneous, breast cancer prognosis and treatment response are difficult to predict with the current prognostic histoclinical parameters. Mammary oncogenesis remains poorly understood. DNA array technology allows the simultaneous analysis of the mRNA expression levels of thousands of genes in biological samples. Applied to breast tumours, expression profiles will boost our knowledge of oncogenesis, will offer new potential therapeutic targets and new prognostic and predictive markers. Today, the most accessible approach for academic research teams is that of Nylon DNA arrays with radioactive detection, which in addition allows profiling of small clinical samples. PMID- 14744111 TI - Osteopontin identified as colon cancer tumor progression marker. AB - Identifying molecular markers for colon cancer is a top priority. Using a pooled sample approach with Affymetrix GeneChip technology, we assayed colon cancers derived from a series of clinical stages to identify molecular markers of potential prognostic value. Of 12000 genes assessed, osteopontin emerged as the leading candidate tumor progression marker. Osteopontin is a secreted glycoprotein known to bind integrins and CD44. Its actual molecular function remains elusive but its increased expression correlates strongly with tumor progression. PMID- 14744112 TI - Regulation of dendritic cell subsets by NKT cells. AB - NKT cells expressing both invariant TCRs and NK cell receptors are an important regulatory cell subset active during initiation of innate immune responses. They are involved in a wide variety of immune responses, but the molecular details of their regulatory action are unknown. Transcriptional profiling has been used for analysis of NKT cell activation profiles, revealing that NKT cells differ from conventional T cells and would be expected to regulate immune responses by controlling dendritic cell activation. PMID- 14744113 TI - In vivo filtering of in vitro expression data reveals MyoD targets. AB - A published set of downstream targets of MyoD defined in a well-controlled in vitro experiment was filtered for relevance to muscle regeneration using a 27 time-point in vivo murine regeneration series. Using interactive hierarchical and Bayes soft clustering, only a minority of the targets defined in vitro can be confirmed in vivo (approximately 50% of induced transcripts, and none of repressed transcripts). This approach provided strong support that 18 targets including of MyoD are biologically relevant during myoblast differentiation. PMID- 14744114 TI - GeneNote: whole genome expression profiles in normal human tissues. AB - A novel data set, GeneNote (Gene Normal Tissue Expression), was produced to portray complete gene expression profiles in healthy human tissues using the Affymetrix GeneChip HG-U95 set, which includes 62 839 probe-sets. The hybridization intensities of two replicates were processed and analyzed to yield the complete transcriptome for twelve human tissues. Abundant novel information on tissue specificity provides a baseline for past and future expression studies related to diseases. The data is posted in GeneNote (http://genecards.weizmann.ac.il/genenote/), a widely used compendium of human genes (http://bioinfo.weizmann.ac.il/genecards). PMID- 14744115 TI - ArrayExpress: a public database of gene expression data at EBI. AB - ArrayExpress is a public repository for microarray-based gene expression data, resulting from the implementation of the MAGE object model to ensure accurate data structuring and the MIAME standard, which defines the annotation requirements. ArrayExpress accepts data as MAGE-ML files for direct submissions or data from MIAMExpress, the MIAME compliant web-based annotation and submission tool of EBI. A team of curators supports the submission process, providing assistance in data annotation. Data retrieval is performed through a dedicated web interface. Relevant results may be exported to ExpressionProfiler, the EBI based expression analysis tool available online (http://www.ebi.ac.uk/arrayexpress). PMID- 14744116 TI - CIBEX: center for information biology gene expression database. AB - We describe the current status of the gene expression database CIBEX (Center for Information Biology gene EXpression database, http://cibex.nig.ac.jp), with a data retrieval system in compliance with MIAME, a standard that the MGED Society has developed for comparing and data produced in microarray experiments at different laboratories worldwide. CIBEX serves as a public repository for a wide range of high-throughput experimental data in gene expression research, including microarray-based experiments measuring mRNA, serial analysis of gene expression (SAGE tags), and mass spectrometry proteomic data. PMID- 14744117 TI - Microarray analysis using bioinformatics analysis audit trails (BAATs). AB - Bioinformatics analysis plays an integrative role in genomics and functional genomics. The ability to conduct quality managed, hypothesis-driven bioinformatics analysis with the plethora of data available is mandatory. Biological interpretation of this data is dependent on versions of databases, programs and the parameters used. Thus, tracking and auditing the analyses process is important. This paper outlines what we term Bioinformatics Analysis Audit Trails (BAATs) and describes YABI, a bioinformatics environment that implements BAATs. YABI can incorporate most bioinformatics tools within the same environment, making it a valuable resource. PMID- 14744118 TI - Application of eVOC: controlled vocabularies for unifying gene expression data. AB - To provide standardised description of gene expression and cross platform querying of databases, we have developed eVOC (http://www.sanbi.ac.za/evoc/), consisting of four orthogonal ontologies which describe Anatomical System, Cell Type, Pathology and Developmental Stage. We have annotated 47 microarray expression data sets and all publicly available human cDNA and SAGE tag libraries. eVOC has been integrated with the public resource EnsMart, which provides linking of transcripts and libraries with expression terms and the human genome sequence (http://www.ensembl.org/Homo_sapiens/martview). PMID- 14744119 TI - Zipf's law and human transcriptomes: an explanation with an evolutionary model. AB - Detailed analysis of human gene expression data reveals several patterns of relationship between transcript frequency and abundance rank. In muscle and liver, organs composed primarily of a homogeneous population of differentiated cells, they obey Zipf's law. In cell lines, epithelial tissue and compiled transcriptome data, only high-rankers deviate from it. We propose an evolutionary process model during which expression level changes stochastically proportionally to its intensity, providing a novel interpretation of transcriptome data and of evolutionary constraints on gene expression. PMID- 14744120 TI - The macro-ethics of genomics to health: the physiome project. AB - Advances in pharmacology and genomics, and their intervention in human biology are beyond our abilities to understand their consequences. Therapeutic intervention in highly complex, non-linear, adaptive biological systems results in some unforeseen and undesirable consequences. To do the most good with the least harm, the information on biological systems should be gathered into databases, and into comprehensive quantitative models that can help to predict the long-range effects of proposed interventions. This is a societal or professional macro-ethical imperative. The Physiome Project helps to meet this imperative via databasing and creating models and tools for large-scale integration. PMID- 14744121 TI - Product patents on human DNA sequences: where do we stand in Europe? AB - In July 1998, the Directive 98/44/EC was adopted by the competent European Community bodies. It should have been implemented into national laws of the member states by the end of July 2000. So far, however, most member states missed that deadline. One reason for resistance and hesitations are concerns about potential negative effects of product patents based on gene sequences. Reasons for this situation are analysed and solutions sought for minimising the prospects of increasing dependencies on dominant patents on genes. PMID- 14744122 TI - Patenting genome research tools and the law. AB - Patenting genes encoding therapeutic proteins was relatively uncontroversial in the early days of biotechnology. Controversy arose in the era of high-throughput DNA sequencing, when gene patents started to look less like patents on drugs and more like patents on scientific information. Evolving scientific and business strategies for exploiting genomic information raised concerns that patents might slow subsequent research. The trend towards stricter enforcement of the utility and disclosure requirements by the patent offices should help clarify the current confusion. PMID- 14744123 TI - The Universal Declaration on the Human Genome and Human Rights. AB - Since 1985, UNESCO studies ethical questions arising in genetics. In 1992, I established the International Bioethics Committee at UNESCO with the mission to draft the Universal Declaration on the Human Genome and Human Rights, which was adopted by UNESCO in 1997 and the United Nations in 1998. The Declaration relates the human genome with human dignity, deals with the rights of the persons concerned by human genome research and provides a reference legal framework for both stimulating the ethical debate and the harmonization of the law worldwide, favouring useful developments that respect human dignity. PMID- 14744124 TI - Patentability of life and ethics. AB - Patent law has relied in part on ethical considerations since its inception in Europe. Such considerations have been introduced more recently in the United States. Whereas the EU Directive on the protection on the occasion of the Human Genome Project of biotechnological inventions was intended to foster economic development in Europe, its implementation is outweighted by controversy about patenting life and commercialization of science. The confusion created must be cleared at the international level through harmonization of patent office policies preventing abusive commercial practices in the absence of inventiveness. PMID- 14744125 TI - Structural elements, mechanism, and evolutionary convergence of Rho protein guanine nucleotide exchange factor complexes. AB - Rho GTPases act as key regulators of cellular biochemistry by determining the timing, direction, and amplitude of signal transduction in a number of important pathways. The rate of activation of a GTPase-controlled reaction is limited by the rate of GTP binding to the Rho protein, and this, in turn, depends on the rate that GDP dissociates from the GTPase. The latter is controlled by the action of guanine nucleotide exchange factors (GEFs) that catalyze GDP-GTP exchange by increasing the rate of GDP dissociation. Here, the recently reported structural information for Rho GTPase-GEF complexes and the molecular basis for the specificity of their interactions are discussed. Underscoring the importance of regulating the Rho GTPase activation pathway, genetically unrelated proteins have evolved which complement or mimic the Dbl homology-Pleckstrin homology (DH-PH) domain-containing family of proteins in their ability to catalyze GDP-GTP exchange. In particular, the structure of the mammalian Cdc42 protein bound to the SopE protein from Salmonella typhimurium illustrates how two unrelated protein folds are able to carry out guanine nucleotide exchange by a remarkably similar mechanism. It will be interesting to see if this conservation of mechanism extends to a newly recognized class of GEFs related to the DOCK180 family. PMID- 14744126 TI - Tazobactam forms a stoichiometric trans-enamine intermediate in the E166A variant of SHV-1 beta-lactamase: 1.63 A crystal structure. AB - Many pathogenic bacteria develop antibiotic resistance by utilizing beta lactamases to degrade penicillin-like antibiotics. A commonly prescribed mechanism-based inhibitor of beta-lactamases is tazobactam, which can function either irreversibly or in a transient manner. We have demonstrated previously that the reaction between tazobactam and a deacylation deficient variant of SHV-1 beta-lactamase, E166A, could be followed in single crystals using Raman microscopy [Helfand, M. S., et al. (2003) Biochemistry 42, 13386-13392]. The Raman data show that maximal populations of an enamine-like intermediate occur 20 30 min after "soaking in" has commenced. By flash-freezing crystals in this time frame, we were able to trap the enamine species. The resulting 1.63 A resolution crystal structure revealed tazobactam covalently bound in the trans-enamine intermediate state with close to 100% occupancy in the active site. The Raman data also indicated that tazobactam forms a larger population of enamine than sulbactam or clavulanic acid does and that tazobactam's intermediate is also the most long-lived. The crystal structure provides a rationale for this finding since only tazobactam is able to form favorable intra- and intermolecular interactions in the active site that stabilize this trans-enamine intermediate. These interactions involve both the sulfone and triazolyl groups that distinguish tazobactam from clavulanic acid and sulbactam, respectively. The observed stabilization of the transient intermediate of tazobactam is thought to contribute to tazobactam's superior in vitro and in vivo clinical efficacy. Understanding the structural details of differing inhibitor effectiveness can aid the design of improved mechanism-based beta-lactamase inhibitors. PMID- 14744127 TI - Family of cytochrome c7-type proteins from Geobacter sulfurreducens: structure of one cytochrome c7 at 1.45 A resolution. AB - The structure of a cytochrome c(7) (PpcA) from Geobacter sulfurreducens was determined by X-ray diffraction at 1.45 A resolution; the R factor is 18.2%. The protein contains a three-heme core that is surrounded by 71 amino acid residues. An unusual feature of this cytochrome is that it has 17 lysine residues, but only nine hydrophobic residues that are larger than alanine. The details of the structure are described and compared with those of cytochrome c(7) from Desulfuromonas acetoxidans and with cytochromes c(3). The two cytochrome c(7) molecules have sequences that are 46% identical, but the arrangements of the hemes in the two structures differ; the rms deviation of all alpha-carbons is 2.5 A. These cytochromes can reduce various metal ions. The reduction site of the chromate ion in D. acetoxidans is occupied by a sulfate ion in the crystal structure of PpcA. We identified four additional homologues of cytochrome c(7) in the G. sulfurreducens genome and three polymers of c(7)-type domains. Of the polymers, two have four repeats and one has nine repeats. On the basis of sequence alignments, one of the hemes in each of the cytochrome c(7)-type domains does not have the bis-histidine coordination. The packing of the molecules in the crystal structure of PpcA suggests that the polymers have an elongated conformation and might form a "nanowire". PMID- 14744128 TI - Nonrandom structure in the urea-unfolded Escherichia coli outer membrane protein X (OmpX). AB - On the basis of sequence-specific resonance assignments for the complete polypeptide backbone and most of the amino acid side chains by heteronuclear nuclear magnetic resonance (NMR) spectroscopy, the urea-unfolded form of the outer membrane protein X (OmpX) from Escherichia coli has been structurally characterized. (1)H-(1)H nuclear Overhauser effects (NOEs), dispersion of the chemical shifts, amide proton chemical shift temperature coefficients, amide proton exchange rates, and (15)N[(1)H]-NOEs show that OmpX in 8 M urea at pH 6.5 is globally unfolded, but adopts local nonrandom conformations in the polypeptide segments of residues 73-82 and 137-145. For these two regions, numerous medium range and longer-range NOEs were observed, which were used as the input for structure calculations of these polypeptide segments with the program DYANA. The segment 73-82 forms a quite regular helical structure, with only loosely constrained amino acid side chains. In the segment 137-145, the tryptophan residue 140 forms the core of a small hydrophobic cluster. Both nonrandom structures are present with an abundance of about 25% of the protein molecules. The sequence-specific NMR assignment and the physicochemical characterization of urea-denatured OmpX presented in this paper are currently used as a platform for investigations of the folding mechanism of this integral membrane protein. PMID- 14744129 TI - Crystal structure of haloalkane dehalogenase LinB from Sphingomonas paucimobilis UT26 at 0.95 A resolution: dynamics of catalytic residues. AB - We present the structure of LinB, a 33-kDa haloalkane dehalogenase from Sphingomonas paucimobilis UT26, at 0.95 A resolution. The data have allowed us to directly observe the anisotropic motions of the catalytic residues. In particular, the side-chain of the catalytic nucleophile, Asp108, displays a high degree of disorder. It has been modeled in two conformations, one similar to that observed previously (conformation A) and one strained (conformation B) that approached the catalytic base (His272). The strain in conformation B was mainly in the C(alpha)-C(beta)-C(gamma) angle (126 degrees ) that deviated by 13.4 degrees from the "ideal" bond angle of 112.6 degrees. On the basis of these observations, we propose a role for the charge state of the catalytic histidine in determining the geometry of the catalytic residues. We hypothesized that double-protonation of the catalytic base (His272) reduces the distance between the side-chain of this residue and that of the Asp108. The results of molecular dynamics simulations were consistent with the structural data showing that protonation of the His272 side-chain nitrogen atoms does indeed reduce the distance between the side-chains of the residues in question, although the simulations failed to demonstrate the same degree of strain in the Asp108 C(alpha)-C(beta)-C(gamma) angle. Instead, the changes in the molecular dynamics structures were distributed over several bond and dihedral angles. Quantum mechanics calculations on LinB with 1-chloro-2,2-dimethylpropane as a substrate were performed to determine which active site conformations and protonation states were most likely to result in catalysis. It was shown that His272 singly protonated at N(delta)(1) and Asp108 in conformation A gave the most exothermic reaction (DeltaH = -22 kcal/mol). With His272 doubly protonated at N(delta)(1) and N(epsilon)(2), the reactions were only slightly exothermic or were endothermic. In all calculations starting with Asp108 in conformation B, the Asp108 C(alpha)-C(beta)-C(gamma) angle changed during the reaction and the Asp108 moved to conformation A. The results presented here indicate that the positions of the catalytic residues and charge state of the catalytic base are important for determining reaction energetics in LinB. PMID- 14744130 TI - Structure of Toxoplasma gondii LDH1: active-site differences from human lactate dehydrogenases and the structural basis for efficient APAD+ use. AB - While within a human host the opportunistic pathogen Toxoplasma gondii relies heavily on glycolysis for its energy needs. Lactate dehydrogenase (LDH), the terminal enzyme in anaerobic glycolysis necessary for NAD(+) regeneration, therefore represents an attractive therapeutic target. The tachyzoite stage lactate dehydrogenase (LDH1) from the parasite T. gondii has been crystallized in apo form and in ternary complexes containing NAD(+) or the NAD(+)-analogue 3 acetylpyridine adenine dinucleotide (APAD(+)) and sulfate or the inhibitor oxalate. Comparison of the apo and ternary models shows an active-site loop that becomes ordered upon substrate binding. This active-site loop is five residues longer than in most LDHs and necessarily adopts a different conformation. While loop isomerization is fully rate-limiting in prototypical LDHs, kinetic data suggest that LDH1's rate is limited by chemical steps. The importance of charge neutralization in ligand binding is supported by the complexes that have been crystallized as well as fluorescence quenching experiments performed with ligands at low and high pH. A methionine that replaces a serine residue and displaces an ordered water molecule often seen in LDH structures provides a structural explanation for reduced substrate inhibition. Superimposition of LDH1 with human muscle- and heart-specific LDH isoforms reveals differences in residues that line the active site that increase LDH1's hydrophobicity. These differences will aid in designing inhibitors specific for LDH1 that may be useful in treating toxoplasmic encephalitis and other complications that arise in immune-compromised individuals. PMID- 14744132 TI - Solution structure of a CCHHC domain of neural zinc finger factor-1 and its implications for DNA binding. AB - The structure of a CCHHC zinc-binding domain from neural zinc finger factor-1 (NZF-1) has been determined in solution though the use of NMR methods. This domain is a member of a family of domains that have the Cys-X(4)-Cys-X(4)-His X(7)-His-X(5)-Cys consensus sequence. The structure determination reveals a novel fold based around a zinc(II) ion coordinated to three Cys residues and the second of the two conserved His residues. The other His residue is stacked between the metal-coordinated His residue and a relatively conserved aromatic residue. Analysis of His to Gln sequence variants reveals that both His residues are required for the formation of a well-defined structure, but neither is required for high-affinity metal binding at a tetrahedral site. The structure suggests that a two-domain protein fragment and a double-stranded DNA binding site may interact with a common two-fold axis relating the two domains and the two half sites of the DNA-inverted repeat. PMID- 14744131 TI - Solution structure and functional characterization of SGTx1, a modifier of Kv2.1 channel gating. AB - SGTx1 is a peptide toxin isolated from the venom of the spider Scodra griseipes that has been shown to inhibit outward K(+) currents in rat cerebellar granule neurons. Although its amino acid sequence is known to be highly (76%) homologous with that of hanatoxin (HaTx), a well-characterized modifier of Kv2.1 channel gating, the structural and functional characteristics of SGTx1 remain largely unknown. Here we describe the NMR solution structure of SGTx1, the mechanism of its interaction with Kv2.1 channels, and its effect on channel activity once bound. The NMR structure of SGTx1 contains a molecular fold closely resembling the "inhibitor cystine knot" of HaTx, which is composed of an antiparallel beta sheet and four chain reversals stabilized by three disulfide bonds. Functionally, SGTx1 reversibly inhibited K(+) currents in oocytes expressing Kv2.1 channels. Moreover, generation of steady-state activation curves showed that, consistent with other gating modifiers, SGTx1 acted by shifting the activation of the channel to more depolarized voltages. Thus, the surface profile and mechanism of action of SGTx1 are similar to those of HaTx. Still, detailed comparison of SGTx1 with HaTx revealed differences in binding affinity and conformational homogeneity that result from differences in the charge distribution at the binding surface and in the amino acid composition of the respective beta-hairpin structures in the peptides. PMID- 14744133 TI - NMR mapping of the HIV-1 Tat interaction surface of the KIX domain of the human coactivator CBP. AB - Tat is required for the expression of the HIV-1 genome. HIV-1 Tat interacts with the human transcriptional coactivator and acetyltransferase CREB-binding protein (CBP) via the KIX domain of CBP. Chemical shift perturbation mapping with nuclear magnetic resonance spectroscopy was used to identify the surface of human KIX that interacts with Tat. It was found that Tat binds to the c-Jun/MLL/Tax binding surface of KIX, as opposed to the CREB binding site. The results provide new insight into the molecular basis of the assembly of protein complexes involving p300/CBP and Tat during HIV gene expression. PMID- 14744134 TI - Dynamics and ligand-induced solvent accessibility changes in human retinoid X receptor homodimer determined by hydrogen deuterium exchange and mass spectrometry. AB - Receptors for retinoic acid act as ligand activated transcription factors. The three-dimensional structure of the retinoid X receptor (RXR) ligand binding domain has been determined, but little information is available concerning the properties of the protein in solution. Hydrogen/deuterium exchange followed by electrospray ionization mass spectrometry was used to probe the solution conformation of the recombinant human RXRalpha homodimer ligand binding domain in the presence and absence of 9-cis-retinoic acid (9-cis-RA). Within the experimental time domain (0.25-180 min), about 20 amide hydrogens showed decreased exchange rates in the presence of saturating concentrations of 9-cis-RA as compared to those found for the homodimer in the absence of ligand. Most of the amides were located in peptides derived from regions of the protein shown by the X-ray structure to interact with the bound ligand: the amino termini of helices 3 and 9, the two beta sheets, helix 8, the H8-H9 loop, and the carboxyl terminus of helix 11. Unexpectedly, protection was also observed in peptides derived from helices 7, 10, 11, and the H7-H8 and H10-H11 loops, regions that are not directly in contact with bound 9-cis-RA. These results suggest that the binding of ligand results in additional effects on the conformation or dynamics of the homodimer in solution as compared to those observed for the X-ray structure. Overall, the change in deuterium exchange induced by the binding of 9 cis-RA correlated reasonably well with changes in hydrogen bonding, residue depth, and/or solvent accessibility predicted from the crystal structure. PMID- 14744136 TI - Cysteine-scanning mutagenesis of transmembrane segment 1 of glucose transporter GLUT1: extracellular accessibility of helix positions. AB - Transmembrane segment 1 of the cysteine-less GLUT1 glucose transporter was subjected to cysteine-scanning mutagenesis. The majority of single-cysteine mutants were functional transporters, as assessed by 2-deoxy-d-glucose uptake or 3-O-methyl-d-glucose transport. Substitution of cysteine for Leu-21, Gly-22, Ser 23, Gln-25, and Gly-27, however, led to uptake rates that were less than 10% of that of the nonmutated cysteine-less GLUT1. NEM, a membrane-permeable agent, was used to identify positions that are sensitive to transport alteration by sulfhydryl reagents, whereas uptake modification by the membrane-impermeant pCMBS indicated accessibility to water-soluble solutes from the external cell environment. Twelve of the 21 single-cysteine mutants were significantly (p < 0.01) affected by NEM, and on the basis of this sensitivity, four positions were identified by pCMBS to form a water-accessible surface within helix 1. The pCMBS sensitive positions are localized at the exofacial C-terminal end along a circumference of the helix. PMID- 14744137 TI - Investigating the importance of the flexible hinge in caerin 1.1: solution structures and activity of two synthetically modified caerin peptides. AB - Caerin 1.1 is a potent broad-spectrum antibacterial peptide isolated from a number of Australian frogs of the Litoria genus. In membrane-like media, this peptide adopts two alpha-helices, separated by a flexible hinge region bounded by Pro15 and Pro19. Previous studies have suggested that the hinge region is important for effective orientation of the two helices within the bacterial cell membrane, resulting in lysis via the carpet mechanism. To evaluate the importance of the two Pro residues, they were replaced with either Ala or Gly. The antibacterial activity of these two peptides was tested, and their three dimensional structures were determined using two-dimensional NMR spectroscopy and restrained molecular dynamics calculations. The resulting structures indicate that the central hinge angle decreases significantly upon replacement of the Pro residues with Gly and to a further extent with Ala. This trend was mirrored by a corresponding decrease in antibiotic activity, further exemplifying the necessity of the hinge in caerin 1.1 and related peptides. In a broader context, the use of Pro, Gly, and Ala variants of caerin 1.1 has enabled the relationship between conformational flexibility and activity to be directly investigated in a systematic manner. PMID- 14744135 TI - Apoptosis of insulin-secreting cells induced by endoplasmic reticulum stress is amplified by overexpression of group VIA calcium-independent phospholipase A2 (iPLA2 beta) and suppressed by inhibition of iPLA2 beta. AB - The death of insulin-secreting beta-cells that causes type I diabetes mellitus (DM) occurs in part by apoptosis, and apoptosis also contributes to progressive beta-cell dysfunction in type II DM. Recent reports indicate that ER stress induced apoptosis contributes to beta-cell loss in diabetes. Agents that deplete ER calcium levels induce beta-cell apoptosis by a process that is independent of increases in [Ca(2+)](i). Here we report that the SERCA inhibitor thapsigargin induces apoptosis in INS-1 insulinoma cells and that this is inhibited by a bromoenol lactone (BEL) inhibitor of group VIA calcium-independent phospholipase A(2) (iPLA(2)beta). Overexpression of iPLA(2)beta amplifies thapsigargin-induced apoptosis of INS-1 cells, and this is also suppressed by BEL. The magnitude of thapsigargin-induced INS-1 cell apoptosis correlates with the level of iPLA(2)beta expression in various cell lines, and apoptosis is associated with stimulation of iPLA(2)beta activity, perinuclear accumulation of iPLA(2)beta protein and activity, and caspase-3-catalyzed cleavage of full-length 84 kDa iPLA(2)beta to a 62 kDa product that associates with nuclei. Thapsigargin also induces ceramide accumulation in INS-1 cells, and this response is amplified in cells that overexpress iPLA(2)beta. These findings indicate that iPLA(2)beta participates in ER stress-induced apoptosis, a pathway that promotes beta-cell death in diabetes. PMID- 14744138 TI - A positive charge at position 33 of thioredoxin primarily affects its interaction with other proteins but not redox potential. AB - Oxidoreductases of the thioredoxin superfamily possess the C-X-X-C motif. The redox potentials vary over a wide range for these proteins. A crucial determinant of the redox potential has been attributed to the variation of the X-X dipeptide. Here, we substitute Lys for Gly at the first X of Escherichia coli thioredoxin to investigate how a positive charge would affect the redox potential. The substitution does not affect the protein's redox potential. The equilibrium constant obtained from pairwise reaction between the mutant and wild-type proteins equals 1.1, indicating that the replacement does not significantly affect the thiol-disulfide redox equilibrium. However, the catalytic efficiency of thioredoxin reductase on the G33K mutant decreases approximately 2.8 times compared to that of the wild type. The mutation mainly affects K(m), with little effect on k(cat). The mutation also inhibits thioredoxin's ability to reduce insulin disulfide by approximately one-half. Whether the mutant protein supports the growth of phages T3/7 and f1 was tested. The efficiency of plating (EOP) of T3/7 on the mutant strain decreases 5 times at 37 degrees C and 3 x 10(4) times at 42 degrees C relative to that of the wild-type strain, suggesting that interaction between phage gene 5 protein and thioredoxin is hindered. The mutation also reduces the EOP of phage f1 by 8-fold at 37 degrees C and 1.5-fold at 42 degrees C. The global structure of the mutant protein does not change when studied by CD and fluorescence spectra. Therefore, G33K does not significantly affect the overall structure or redox potential of thioredoxin, but primarily interferes with its interaction with other proteins. Together with the G33D mutation, the overall results show that a charged residue at the first X has a greater influence on the molecular interaction of the protein than the redox potential. PMID- 14744140 TI - G-1:C73 recognition by an arginine cluster in the active site of Escherichia coli histidyl-tRNA synthetase. AB - Aminoacylation of a transfer RNA (tRNA) by its cognate aminoacyl-tRNA synthetase relies upon the recognition of specific nucleotides as well as conformational features within the tRNA by the synthetase. In Escherichia coli, the aminoacylation of tRNA(His) by histidyl-tRNA synthetase (HisRS) is highly dependent upon the recognition of the unique G-1:C73 base pair and the 5' monophosphate. This work investigates the RNA-protein interactions between the HisRS active site and these critical recognition elements. A homology model of the tRNA(His)-HisRS complex was generated and used to design site-specific mutants of possible G-1:C73 contacts. Aminoacylation assays were performed with these HisRS mutants and N-1:C73 tRNA(His) and microhelix(His) variants. Complete suppression of the negative effect of 5'-phosphate deletion by R123A HisRS, as well as the increased discrimination of Q118E HisRS against a 5'-triphosphate, suggests a possible interaction between the 5'-phosphate and active-site residues Arg123 and Gln118 in which these residues create a sterically and electrostatically favorable pocket for the binding of the negatively charged phosphate group. Additionally, a network of interactions appears likely between G 1 and Arg116, Arg123, and Gln118 because mutation of these residues significantly reduced the sensitivity of HisRS to changes at G-1. Our studies also support an interaction previously proposed between Gln118 and C73. Defining the RNA-protein interactions critical for efficient aminoacylation by E. coli HisRS helps to further characterize the active site of this enzyme and improves our understanding of how the unique identity elements in the acceptor stem of tRNA(His) confer specificity. PMID- 14744139 TI - Discrimination between different methylation states of chemotaxis receptor Tar by receptor methyltransferase CheR. AB - Bacterial chemotaxis receptors are posttranslationally modified by carboxyl methylation of specific glutamate residues within their cytoplasmic domains. This highly regulated, reversible modification counterbalances the signaling effects of ligand binding and contributes to adaptation. On the basis of the crystal structure of the gamma-glutamyl methyltransferase CheR, we have postulated that positively charged residues in helix alpha2 in the N-terminal domain of the enzyme may be complementary to the negatively charged methylation region of the methyltransferase substrates, the bacterial chemotaxis receptors. Several altered CheR proteins, in which positively charged arginine or lysine residues were substituted with alanines, were constructed and assayed for their methylation activities toward wild-type receptor and a series of receptor variants containing different glutamates available for methylation. One of the CheR mutant proteins (Arg53Ala) showed significantly lower activity toward all receptor constructs, suggesting that Arg53 may play a general role in catalysis of methyl transfer. The rest of the mutant proteins exhibited different patterns of relative methylation rates toward different receptor substrates, indicating specificity, probably through interaction of CheR with the receptor at sites distal to the specific site of methylation. The findings imply complementarity between positively charged residues of the alpha2 helix of CheR and the negatively charged glutamates of the receptor. It is likely that this complementarity is involved in discriminating different methylation states of the receptors. PMID- 14744141 TI - Role of the active site cysteine of DpgA, a bacterial type III polyketide synthase. AB - DpgA is a bacterial type III polyketide synthase (PKS) that decarboxylates and condenses four malonyl-CoA molecules to produce 3,5-dihydroxyphenylacetyl-CoA (DPA-CoA) in the biosynthetic pathway to 3,5-dihydroxyphenylglycine, a key nonproteinogenic residue in the vancomycin family of antibiotics. DpgA has the conserved catalytic triad of Cys/His/Asn typical of type III PKS enzymes, and has been assumed to use Cys160 as the catalytic nucleophile to create a series of elongating acyl-S-enzyme intermediates prior to the C(8) to C(3) cyclization step. Incubation of purified DpgA with [(14)C]-malonyl-CoA followed by acid quench during turnover leads to accumulation of 10-15% of the DpgA molecules covalently acylated. Mutation of the active site Cys160 to Ala abrogated detectable covalent acylation, but the C160A mutant retained 50% of the V(max) for DPA-CoA formation, with a k(cat) still at 0.5 catalytic turnovers/min. For comparison, a C190A mutant retained wild-type activity, while the H296A mutant, in which the side chain of the presumed catalytic His is removed, had a 6-fold drop in k(cat). During turnover, purified DpgA produced 1.2 equivalents of acetyl CoA for each DPA-CoA, indicating 23% uncoupled decarboxylation competing with condensative C-C coupling. The C160A mutant showed an increased partition ratio for malonyl-CoA decarboxylation to acetyl-CoA vs condensation to DPA-CoA, reflecting more uncoupling in the mutant enzyme. The Cys-to-Ala mutant thus shows the unexpected result that, when the normal acyl-S-enzyme mechanism for this type III PKS elongation/cyclization catalyst is removed, it can still carry out the regioselective construction of the eight-carbon DPA-CoA skeleton with surprising efficiency. PMID- 14744142 TI - Selection of human cytochrome P450 1A2 mutants with enhanced catalytic activity for heterocyclic amine N-hydroxylation. AB - Cytochrome P450 (P450) 1A2 is the major enzyme involved in the metabolism of 2 amino-3,5-dimethylimidazo[4,5-f]quinoline (MeIQ) and other heterocyclic arylamines and their bioactivation to mutagens. Random mutant libraries of human P450 1A2, in which mutations were made throughout the entire open reading frame, were screened with Escherichia coli DJ3109pNM12, a strain designed to bioactivate MeIQ and detect mutagenicity of the products. Mutant clones with enhanced activity were confirmed using quantitative measurement of MeIQ N-hydroxylation. Three consecutive rounds of random mutagenesis and screening were performed and yielded a highly improved P450 1A2 mutant, SF513 (E225N/Q258H/G437D), with >10 fold increased MeIQ activation based on the E. coli genotoxicity assay and 12 fold enhanced catalytic efficiency (k(cat)/K(m)) in steady-state N-hydroxylation assays done with isolated membrane fractions. SF513 displayed selectively enhanced activity for MeIQ compared to other heterocyclic arylamines. The enhanced catalytic activity was not attributed to changes in any of several individual steps examined, including substrate binding, total NADPH oxidation, or H(2)O(2) formation. Homology modeling based on an X-ray structure of rabbit P450 2C5 suggested that the E225N and Q258H mutations are located in the F-helix and G helix, respectively, and that the G437D mutation is in the "meander" region, apparently rather distant from the substrate. In summary, the approach generated a mutant enzyme with selectively elevated activity for a single substrate, even to the extent of a difference of a single methyl group, and several mutations had interacting roles in the development of the selected mutant protein. PMID- 14744143 TI - Evidence for a direct manganese-oxygen ligand in water binding to the S2 state of the photosynthetic water oxidation complex. AB - The interaction of water with the water oxidizing Mn complex of photosystem II has been investigated using electron spin-echo envelope modulation spectroscopy in the presence of H(2)(17)O. The spectra show interaction of the (17)O with the preparation in the S(2) state induced by 200 K illumination. The modulation is observed only in the center of the multiline spectrum. The inferred hyperfine coupling terms are compatible with water (not hydroxyl) oxygen bound to a particular quasi-axial Mn(III) center in a coupled Mn cluster. PMID- 14744144 TI - Consequences of counterion mutation in sensory rhodopsin II of Natronobacterium pharaonis for photoreaction and receptor activation: an FTIR study. AB - In many retinal proteins the proton transfer from the Schiff base to the counterion represents a functionally important step of the photoreaction. In the signaling state of sensory rhodopsin II from Natronobacterium pharaonis this transfer has already occurred, but in the counterion mutant Asp75Asn it is blocked during all steps of the photocycle. Therefore, the study of the molecular changes during the photoreaction of this mutant should provide a deeper understanding of the activation mechanism, and for this, we have applied time resolved step-scan FTIR spectroscopy. The photoreaction is drastically altered; only red-shifted intermediates are formed with a chromophore strongly twisted around the 14-15 single bond. In addition, the photocycle is shortened by 2 orders of magnitude. Nevertheless, a transition involving only protein changes similar to that of the wild type is observed, which has been correlated with the formation of the signaling state. However, whereas in the wild type this transition occurs in the millisecond range, it is shortened to 200 micros in the mutant. The results are discussed with respect to the altered electrostatic interactions, role of proton transfer, the published 3D structure, and physiological activity. PMID- 14744145 TI - Specific modification of two tryptophans within the nuclear-encoded subunits of bovine cytochrome c oxidase by hydrogen peroxide. AB - Hydrogen peroxide does more than react with the binuclear center of oxidized bovine cytochrome c oxidase and generate the well-characterized "peroxy" and "ferryl" forms. Hydrogen peroxide also inactivates detergent-solubilized cytochrome c oxidase in a time- and concentration-dependent manner. There is a 70 80% decrease of electron-transport activity, peroxidation of bound cardiolipin, modification of two nuclear-encoded subunits (IV and VIIc), and dissociation of approximately 60% of subunits VIa and VIIa. Modification of subunit VIIc and dissociation of subunit VIIa are coupled events that probably are responsible for the inactivation of cytochrome c oxidase. When cytochrome c oxidase is exposed to 500 microM hydrogen peroxide for 30 min at pH 7.4 and room temperature, subunits IV (modified up to 20%) and VIIc (modified up to 70%) each have an increased mass of 16 Da as detected by matrix-assisted laser desorption/ionization time-of flight mass spectrometry and electrospray ionization mass spectrometry. In each case, the increased mass is caused by oxidation of a tryptophan (Trp19 within subunit VIIc and Trp48 within subunit IV), almost certainly due to formation of hydroxytryptophan. We conclude that hydrogen peroxide-induced oxidation of tryptophan and cardiolipin proceeds via the binuclear center since both modifications are prevented if the binuclear center is first blocked with cyanide. Bound cardiolipin and oxidized tryptophans are localized relatively far from the binuclear center (30-60 A); therefore, oxidation probably occurs by migration of a free radical generated at the binuclear center to these distal reaction sites. PMID- 14744147 TI - Molecular dynamics simulations and oxidation rates of methionine residues of granulocyte colony-stimulating factor at different pH values. AB - To understand the connection between the conformation of a protein molecule and the oxidation of its methionine residues, we measured the rates of oxidation of methionine residues by H(2)O(2) in granulocyte colony-stimulating factor (G-CSF) as a function of pH and also studied the structural properties of this protein as a function of pH via molecular dynamics simulations. We found that each of the four methionine groups in G-CSF have significant and different rates of oxidation as a function of pH. Moreover, Met(1), in the unstructured N-terminal region, has a rate of oxidation as low as half that of free methionine. The structural properties of G-CSF as a function of pH are evaluated in terms of properties such as hydrogen bonding, deviations from X-ray structure, helical/helical packing, and the atomic covariance fluctuation matrix of alpha-carbons. We found that dynamics (structural fluctuations) are essential in explaining oxidation and that a static picture, such as that resulting from X-ray data, fails in this regard. Moreover, the simulation results also indicate that the solvent-accessible area, traditionally used to measure solvent accessibility of a protein site, of the sulfur atom of methionine residues does not correlate well with the rate of oxidation. Instead, we identified a structural property, average two-shell water coordination number, that correlates well with measured oxidation rates. PMID- 14744146 TI - Relationship between sterol/steroid structure and participation in ordered lipid domains (lipid rafts): implications for lipid raft structure and function. AB - The formation and stability of ordered lipid domains (rafts) in model membrane vesicles were studied using a series of sterols and steroids structurally similar to cholesterol. In one assay, insolubility in Triton X-100 was assessed in bilayers composed of sterol/steroid mixed with dipalmitoylphosphatidylcholine (DPPC), dioleoylphosphatidylcholine, or a 1:1 mixture of these phospholipids. In a second assay fluorescence quenching was used to determine the degree of ordered domain formation in bilayers containing sterol/steroid and a 1:1 mixture of DPPC and a quencher-carrying phosphatidylcholine. Both methods showed that several single modifications of the cholesterol structure weaken, but do not fully abolish, the ability of sterols and steroids to promote ordered domain formation when mixed with DPPC. Some of these modifications included a shift of the double bond from the 5-6 carbons (cholesterol) to 4-5 carbons (allocholesterol), derivatization of the 3-OH (cholesterol methyl ether, cholesteryl formate), and alteration of the 3-hydroxy to a keto group (cholestanone). An oxysterol involved in atherosclerosis, 7-ketocholesterol, formed domains with DPPC that were as thermally stable as those with cholesterol although not as tightly packed as judged by fluorescence anisotropy. It was also found that 7-ketocholesterol has fluorescence quenching properties making it a useful spectroscopic probe. Lathosterol, which has a 7-8 carbon double bond in place of the 5-6 double bond of cholesterol, formed rafts with DPPC that were at least as detergent-resistant as, and even more thermally stable than, rafts containing cholesterol. Because lathosterol is an intermediate in cholesterol biosynthesis, we conclude it is unlikely that sterol biosynthesis continues past lathosterol in order to create a raft-favoring lipid. PMID- 14744148 TI - Substrate preference in phosphatidylserine biosynthesis for docosahexaenoic acid containing species. AB - Neuronal membranes contain high levels of phosphatidylserine (PS) and docosahexaenoic acid (22:6n-3, DHA). In this study, substrate preference in PS synthesis was determined to gain insight on the biochemical basis for concentrating PS in neuronal membranes where 22:6n-3 is highly enriched. We first established an in vitro assay method using unilamellar vesicles (LUV) of deuterium-labeled substrates and reversed-phase HPLC/electrospray ionization (ESI) mass spectrometry. The PS production by the incubation of deuterium-labeled substrate and microsomal fractions was monitored. We found that tissue-specific substrate preference exists in PS synthesis. Microsomes from the cerebral cortex synthesized PS from 18:0,22:6-PC most favorably among the PC substrates tested, followed by 18:0,22:5-PC, resulting in the PC substrate preference in the order of 18:0,22:6 > 18:0,22:5 > 18:0,20:4 = 18:0,18:1. Liver microsomes also preferred 18:0,22:6-PC as the substrate in PS synthesis but did not use 18:0,22:5-PC favorably. The 18:0,22:5-PC species was converted to PS at the similar extent as 18:0,20:4- or 18:0,18:1-PC species in the liver. Both brain and liver microsomes showed a preference for 18:0 over 16:0 as the sn-1 fatty acid. From these data it was deduced that preferential conversion of 18:0,22:6-PC to the corresponding PS species is at least partly responsible for concentrating PS in neuronal tissues where 22:6n-3 is particularly abundant. The distinctive preference for 18:0,22:5 PS observed with brain microsomes may help to maintain PS at a high level in the brain when 22:6n-3 is replaced by 22:5n-3 as in the case of n-3 fatty acid deficiency. PMID- 14744149 TI - A two-module region of the low-density lipoprotein receptor sufficient for formation of complexes with apolipoprotein E ligands. AB - The low-density lipoprotein (LDL) receptor transports two different classes of cholesterol-carrying lipoprotein particles into cells: LDL particles, which contain a single copy of apolipoprotein B-100 (apoB-100), and beta-migrating very low-density lipoprotein (beta-VLDL) particles, which contain multiple copies of apolipoprotein E (apoE). The ligand-binding domain of the receptor lies at its amino-terminal end within seven adjacent LDL-A repeats (LA1-LA7). Although prior work clearly establishes that LA5 is required for high-affinity binding of particles containing apolipoprotein E (apoE), the number of ligand-binding repeats sufficient to bind apoE ligands has not yet been determined. Similarly, uncertainty exists as to whether a single lipid-activated apoE receptor-binding site within a particle is capable of binding to the LDLR with high affinity or whether more than one is required. Here, we establish that the LA4-5 two-repeat pair is sufficient to bind apoE-containing ligands, on the basis of binding studies performed with a series of LDLR-derived "minireceptors" containing up to four repeats. Using single chain multimers of the apoE receptor-binding domain (N apoE), we also show that more than one receptor-binding site in its lipid activated conformation is required to bind to the LDLR with high affinity. Thus, in addition to inducing a conformational change in the structure of N-apoE, lipid association enhances the affinity of apoE for the LDLR in part by creating a multivalent ligand. PMID- 14744150 TI - Characterization of the adenosinetriphosphatase and transport activities of purified cystic fibrosis transmembrane conductance regulator. AB - The cystic fibrosis transmembrane conductance regulator (CFTR) functions in vivo as a cAMP-activated chloride channel. A member of the ATP-binding cassette superfamily of membrane transporters, CFTR contains two transmembrane domains (TMDs), two nucleotide-binding domains (NBDs), and a regulatory (R) domain. It is presumed that CFTR couples ATP hydrolysis to channel gating, and as a first step in addressing this issue directly, we have established conditions for purification of biochemical quantities of human CFTR expressed in Sf9 insect cells. Use of an 8-azido[alpha-(32)P]ATP-binding and vanadate-trapping assay allowed us to devise conditions to preserve CFTR function during purification of a C-terminal His(10)-tagged variant after solubilization with lysophosphatidylglycerol (1%) and diheptanoylphosphatidylcholine (0.3%) in the presence of excess phospholipid. Study of purified and reconstituted CFTR showed that it binds nucleotide with an efficiency comparable to that of P-glycoprotein and that it hydrolyzes ATP at rates sufficient to account for presumed in vivo activity [V(max) of 58 +/- 5 nmol min(-1) (mg of protein)(-1), K(M)(MgATP) of 0.15 mM]. In further work, we found that neither nucleotide binding nor ATPase activity was altered by phosphorylation (using protein kinase A) or dephosphorylation (with protein phosphatase 2B); we also observed inhibition (approximately 40%) of ATP hydrolysis by reduced glutathione but not by DTT. To evaluate CFTR function as an anion channel, we introduced an in vitro macroscopic assay based on the equilibrium exchange of proteoliposome-entrapped radioactive tracers. This revealed a CFTR-dependent transport of (125)I that could be inhibited by known chloride channel blockers; no significant CFTR-dependent transport of [alpha-(32)P]ATP was observed. We conclude that heterologous expression of CFTR in Sf9 cells can support manufacture and purification of fully functional CFTR. This should aid in further biochemical characterization of this important molecule. PMID- 14744151 TI - Binding of the b-subunit in the ATP synthase from Escherichia coli. AB - The rotary mechanism of ATP synthase requires a strong binding within stator subunits. In this work we studied the binding affinity of the b-subunit to F(1) ATPase of Escherichia coli. The dimerization of the truncated b-subunit without amino acids 1-33, b(34-156)T62C, was investigated by analytical ultracentrifugation, resulting in a dissociation constant of 1.8 microM. The binding of b-subunit monomeric and dimeric forms to the isolated F(1) part was investigated by fluorescence correlation spectroscopy and steady-state fluorescence. The mutants b(34-156)T62C and EF(1)-gammaT106C were labeled with several fluorophores. Fluorescence correlation spectroscopy was used to measure translational diffusion times of the labeled b-subunit, labeled F(1), and a mixture of the labeled b-subunit with unlabeled F(1). Data analysis revealed a dissociation constant of 0.2 nM of the F(1)b(2) complex, yielding a Gibbs free energy of binding of DeltaG(o)= -55 kJ mol(-1). In steady-state fluorescence resonance energy transfer (FRET) measurements it was found that binding of the b subunit to EF(1)-gammaT106C-Alexa488 resulted in a fluorescence decrease of one third of the initial FRET donor fluorescence intensity. The decrease of fluorescence was measured as a function of b-concentration, and data were described by a model including equilibria for dimerization of the b-subunit and binding of b and b(2) to F(1). For a quantitative description of fluorescence decrease we used two different models: the binding of the first and the second b subunit causes the same fluorescence decrease (model 1) or only the binding of the first b-subunit causes fluorescence decrease (model 2). Data evaluation revealed a dissociation constant for the F(1)b(2) complex of 0.6 nM (model 1) or 14 nM (model 2), giving DeltaG(o)= -52 kJ mol(-1) and DeltaG(o)= -45 kJ mol(-1), respectively. The maximal DeltaG observed for ATP synthesis in cells is approximately DeltaG= 55 kJ mol(-1). Therefore, the binding energy of the b subunit seems to be too low for models in which the free energy for ATP synthesis is accumulated in the elastic strain between rotor and stator subunits and then transduced to the catalytic site in one single step. Models in which energy transduction takes place in at least two steps are favored. PMID- 14744152 TI - Rates of elementary catalytic steps for different metal forms of the family II pyrophosphatase from Streptococcus gordonii. AB - Soluble inorganic pyrophosphatases (PPases) form two nonhomologous families, denoted I and II, that have similar active-site structures but different catalytic activities and metal cofactor specificities. Family II PPases, which are often found in pathogenic bacteria, are more active than family I PPases, and their best cofactor is Mn(2+) rather than Mg(2+), the preferred cofactor of family I PPases. Here, we present results of a detailed kinetic analysis of a family II PPase from Streptococcus gordonii (sgPPase), which was undertaken to elucidate the factors underlying the different properties of family I and II PPases. We measured rates of PP(i) hydrolysis, PP(i) synthesis, and P(i)/water oxygen exchange catalyzed by sgPPase with Mn(2+), Mg(2+), or Co(2+) in the high affinity metal-binding site and Mg(2+) in the other sites, as well as the binding affinities for several active-site ligands (metal cofactors, fluoride, and P(i)). On the basis of these data, we deduced a minimal four-step kinetic scheme and evaluated microscopic rate constants for all eight relevant reaction steps. Comparison of these results with those obtained previously for the well-known family I PPase from Saccharomyces cerevisiae (Y-PPase) led to the following conclusions: (a) catalysis by sgPPase does not involve the enzyme-PP(i) complex isomerization known to occur in family I PPases; (b) the values of k(cat) for the magnesium forms of sgPPase and Y-PPase are similar because of similar rates of bound PP(i) hydrolysis and product release; (c) the marked acceleration of sgPPase catalysis in the presence of Mn(2+) and Co(2+) results from a combined effect of these ions on bound PP(i) hydrolysis and P(i) release; (d) sgPPase exhibits lower affinity for both PP(i) and P(i); and (e) sgPPase and Y-PPase exhibit similar values of k(cat)/K(m), which characterizes the PPase efficiency in vivo (i.e., at nonsaturating PP(i) concentrations). PMID- 14744153 TI - Kinetic characterization of yeast pyruvate carboxylase isozyme Pyc1 and the Pyc1 mutant, C249A. AB - The yeast Pyc1 isoform of pyruvate carboxylase has been further characterized and shown to differ from the Pyc2 isoform in its K(a) for K(+) activation. Pyc1 differs from chicken liver pyruvate carboxylase in the lack of effect of acetyl CoA on ADP phosphorylation by carbamoyl phosphate, which may be a result of differences in the loci of action of the effector between the two enzymes. Solvent D(2)O isotope effects have been measured with Pyc1 on the full pyruvate carboxylation reaction, the ATPase reaction in the absence of pyruvate, and the carbamoyl phosphate-ADP phosphorylation reaction for the first time for pyruvate carboxylase. Proton inventories indicate that the measured isotope effects are due to a single proton transfer step in the reaction. The inverse isotope effects observed in all reactions suggest that the proton transfer step converts the enzyme from an inactive to an active form. Kinetic measurements on the C249A mutant enzyme suggest that C249 is involved in the binding and action of enzyme activators K(+) and acetyl-CoA. C249 is not involved in ATP binding as was observed for the corresponding residue in the biotin carboxylase subunit of Escherichia coli acetyl-CoA carboxylase, nor is it directly responsible for the measured inverse (D)(k(cat)/K(m)) isotope effects. The size of the inverse isotope effects indicates that they may result from formation of a low-barrier hydrogen bond. Modification of the wild type and C249A mutant with o phthalaldehyde suggests that C249 is involved in isoindole formation but that the modification of this residue is not directly responsible for the accompanying major loss of enzyme activity. PMID- 14744154 TI - Reactions of serine palmitoyltransferase with serine and molecular mechanisms of the actions of serine derivatives as inhibitors. AB - Serine palmitoyltransferase (SPT) is a key enzyme in sphingolipid biosynthesis and catalyzes the decarboxylative condensation of L-serine and palmitoyl coenzyme A to 3-ketodihydrosphingosine. We have succeeded in the overproduction of a water soluble homodimeric SPT from Sphingomonas paucimobilis EY2395(T) in Escherichia coli. The recombinant SPT showed the characteristic absorption and circular dichroism spectra derived from its coenzyme pyridoxal 5'-phosphate. On the basis of the spectral changes of SPT, we have analyzed the reactions of SPT with compounds related to L-serine and product, and showed the following new aspects: First, we analyzed the binding of L-serine and 3-hydroxypropionate and found that the spectral change in SPT by the substrate is caused by the formation of an external aldimine intermediate and not by the formation of the Michaelis complex. Second, various serine analogues were also examined; the data indicated that the alpha-carboxyl group of L-serine was quite important for substrate recognition by SPT. Third, we focused on a series of SPT inhibitors, which have been used as convenient tools to study the cell responses caused by sphingolipid depletion. The interaction of SPT with myriocin suggested that such product-related compounds would strongly and competitively inhibit enzyme activity by forming an external aldimine in the active site of the enzyme. Beta-chloro-L-alanine and L cycloserine were found to generate characteristic PLP-adducts that produced inactivation of SPT in an irreversible manner. The detailed mechanisms for the SPT inactivation were discussed. This is the first analysis of the inhibition mechanisms of SPT by these compounds, which will provide an enzymological basis for the interpretation of the results from cell biological experiments. PMID- 14744155 TI - Propeptide does not act as an intramolecular chaperone but facilitates protein disulfide isomerase-assisted folding of a conotoxin precursor. AB - Conotoxins comprise a large and diverse group of peptide neurotoxins derived from Conus snail venoms; most contain multiple disulfide bonds. The conotoxin precursors consist of three distinct domains: the N-terminal signal sequence, an intervening propeptide region, and the C-terminal mature conotoxin. Formation of the native disulfide bonds during the oxidative folding of conotoxins is a prerequisite for their proper biological function, but in numerous in vitro folding experiments with mature conotoxins, a lack of specificity in formation of the native Cys-Cys connectivities is observed. The mechanisms that ensure that the native disulfide bonds are formed in venom ducts during biosynthesis remain unknown. To evaluate whether the propeptide could potentially function as an intramolecular chaperone, we studied the oxidative folding of a conotoxin precursor, pro-GI, belonging to the alpha-conotoxin family. Our results indicate that the propeptide sequence did not directly contribute to folding kinetics and thermodynamics. However, we found that the propeptide region of pro-GI played an important role when oxidative folding was catalyzed by protein disulfide isomerase (PDI). The PDI-assisted reaction was more efficient during the early folding in the context of the propeptide sequence (pro-GI), as compared to that of the mature conotoxin (alpha-GI). Taken together, our results suggest for the first time that the propeptide region may play a role in the PDI-catalyzed oxidative folding of conotoxin precursors. PMID- 14744156 TI - Direct comparison of A- and T-strand minor groove interactions in DNA curvature at A tracts. AB - To investigate the relative contributions of minor-groove electrostatic interactions in the mechanism of A-tract DNA curvature, we carried out experiments with modified DNA bases in both strands of the tract. We employed 3 deazaadenine nucleoside (D), which lacks the adenine N3 nitrogen in the minor groove and thus cannot act as an electron donor, as well as difluorotoluene (F), a nonpolar thymine mimic. The effects of these analogues in A-tract curvature were quantified using ligation ladder gel mobility methods developed by Crothers and by Maher. Through single substitutions of D in A(5) tracts, we found that this analogue results in decreased curvature only when situated toward the 3' end of the tract. This is distinct from the behavior in the T-rich strand where F substitution causes the greatest reductions in curvature toward the 5' end. To test for cooperative pairwise effects, we also studied 10 different D + F double substitutions and found evidence supporting a number of localized cooperative electrostatic interactions but not between the two most sensitive sites in the opposite strands. These results suggest that there are two discrete locations in the A-tract minor groove where electrostatic interactions are important in causing curvature: one near the 5' end of the T-rich strand, and one near the 3' end of the A-rich strand. The results are consistent with an important role of localized cations in the minor groove. Possible effects of groove solvation and stacking at the A-tract junction are also discussed. PMID- 14744157 TI - DNA enzyme generated by a novel single-stranded DNA expression vector inhibits expression of the essential bacterial cell division gene ftsZ. AB - Rapid emergence of antibiotic-resistant bacterial pathogens has created urgent demand for the discovery and development of new antibacterial agents directed toward novel targets. Antisense oligodeoxynucleotides (AS-ODN) and their modified forms have been utilized to block gene expression in bacterial cells, showing potential for developing highly specific and efficacious antibacterial agents. In this study, a tetracycline-regulated expression vector was developed for generating single-stranded DNA (ssDNA) of a desired target sequence in bacterial cells. This inducible ssDNA expression vector was tested for producing a DNA enzyme designed to specifically cleave ftsZ mRNA. Our results indicate that the expressed DNA enzyme molecules not only repress ftsZ gene expression and but also inhibit bacterial cell proliferation. Although we believe that the cleavage of ftsZ mRNA by the expressed DNA enzyme molecules is responsible for the inhibitory effects on ftsZ gene expression and bacterial cell proliferation, the antisense mechanism could also be responsible for the biological effects. The ability of this ssDNA expression system to selectively modulate gene expression may provide a powerful strategy in determining the contribution of a given gene product to bacterial growth or pathogenesis and opens a new venue for developing antibacterial agents. PMID- 14744158 TI - Efficient, pH-dependent RNA ligation by the VS ribozyme in trans. AB - The VS ribozyme acts as a very efficient ligase in trans when the 5' cleavage product is prevented from dissociation by an extended helix Ia in the substrate. Provided that the length of this helix is >or=10 bp, the substrate becomes approximately 80% ligated by the ribozyme acting in trans. Most of the nucleotides that have been shown to be important for cleavage are similarly important for ligation, including the critical A756 of the active site. The exception to this is C755. The variant ribozyme C755A has almost normal cleavage activity, whereas the rate of ligation is reduced 70-fold. It is therefore likely that this nucleotide plays a specific role in the organization of the termini of the ligation substrates. We have found that the rate of the trans ligation reaction depends on pH, corresponding to the protonation/deprotonation of a group with a pK(A) of 5.6. A model is suggested whereby the approach to equilibrium is catalyzed by the ribozyme catalyzing the ligation reaction in its deprotonated state (rate 1.05 min(-1)) and the cleavage reaction in its protonated state (rate 0.18 min(-1)). A756 is a candidate for the nucleobase undergoing protonation/deprotonation. PMID- 14744159 TI - NMR spectroscopy of phosphorylated wild-type rhodopsin: mobility of the phosphorylated C-terminus of rhodopsin in the dark and upon light activation. AB - Binding of arrestin to light-activated rhodopsin involves recognition of the phosphorylated C-terminus and several residues on the cytoplasmic surface of the receptor. These sites are in close proximity in dark, unphosphorylated rhodopsin. To address the position and mobility of the phosphorylated C-terminus in the active and inactive receptor, we combined high-resolution solution and solid state NMR spectroscopy of the intact mammalian photoreceptor rhodopsin in detergent micelles as a function of temperature. The (31)P NMR resonance of rhodopsin phosphorylated by rhodopsin kinase at the C-terminal tail was observable with single pulse excitation using magic angle spinning until the sample temperature reached -40 degrees C. Below this temperature, the (31)P resonance broadened and was only observable using cross polarization. These results indicate that the phosphorylated C-terminus is highly mobile above -40 degrees C and immobilized at lower temperature. To probe the relative position of the immobilized phosphorylated C-terminus with respect to the cytoplasmic domain of rhodopsin, (19)F labels were introduced at positions 140 and 316 by the reaction of rhodopsin with 2,2,2-trifluoroethanethiol (TET). Solid state rotational-echo double-resonance (REDOR) NMR was used to probe the internuclear distance between the (19)F and the (31)P-labels. The REDOR technique allows (19)F...(31)P distances to be measured out to approximately 12 A with high resolution, but no significant dephasing was observed in the REDOR experiment in the dark or upon light activation. This result indicates that the distances between the phosphorylated sites on the C-terminus and the (19)F sites on helix 8 (Cys 316) and in the second cytoplasmic loop (Cys140) are greater than 12 A in phosphorylated rhodopsin. PMID- 14744162 TI - Symptoms and circuits, part 3: schizophrenia. AB - Schizophrenia is composed of not only positive symptoms, but also cognitive and affective symptoms that contribute significantly to morbidity. Each of these symptoms may be mediated by a separate and distinct neuronal circuit. PMID- 14744163 TI - Glycogen synthase kinase-3: a target for novel bipolar disorder treatments. AB - The enzyme glycogen synthase kinase-3 (GSK-3) is a direct target of lithium. While originally recognized as an important molecule in a limited number of cellular processes, with unclear significance for the treatment of bipolar disorder, recent evidence suggests it has critically important cellular functions in the adult brain. GSK-3 has an essential role in a number of signaling pathways and regulates the function of a diverse number of proteins, notably transcription factors and cytoskeletal elements. The most important functions of the enzyme in regard to bipolar disorder may be critical effects on cellular resilience and neuronal plasticity. There is tremendous interest in GSK-3 inhibitors as novel therapeutic agents, and selective, small-molecule compounds are rapidly being developed for a broad range of other maladies including diabetes, Alzheimer's disease, stroke, and inflammatory conditions. In this perspectives article, we provide an overview of the molecular targets of lithium, focusing on GSK-3 regulated signaling pathways and the important functions of GSK-3 that may have relevance for the treatment of bipolar disorder. We conclude with a discussion of the GSK-3 inhibitors furthest in development and the clinical trials that may emerge. PMID- 14744164 TI - Impact of age at onset and duration of illness on the expression of comorbidities in obsessive-compulsive disorder. AB - BACKGROUND: Obsessive-compulsive disorder (OCD) patients usually experience comorbidities including tics, trichotillomania, body dysmorphic disorder, and mood and anxiety disorders. The present report verifies how age at onset of obsessive-compulsive symptoms and duration of illness are associated with comorbid diagnoses in OCD patients. METHOD: Psychiatric comorbidity was assessed using a structured clinical interview in 161 consecutive outpatients referred for treatment between 1996 and 2001 who met DSM-IV criteria for OCD. Age at onset and duration of illness were retrospectively assessed by direct interviews. RESULTS: An earlier age at onset of obsessive-compulsive symptoms was associated with tic disorders, while longer illness duration was associated with depressive disorder (major depressive disorder or dysthymia) and social phobia. CONCLUSION: Age at onset and duration of OCD illness are meaningful variables affecting the expression of comorbidities in OCD. Tic disorders and OCD may share common etiologic pathways. Depressive disorders, in contrast, may be secondary complications of OCD. PMID- 14744165 TI - Mental health service utilization by borderline personality disorder patients and Axis II comparison subjects followed prospectively for 6 years. AB - OBJECTIVE: The purpose of this study was to describe the psychiatric treatment received by a well-defined sample of patients with borderline personality disorder and Axis II comparison subjects over 6 years of prospective follow-up. METHOD: 362 inpatients were interviewed about their treatment histories during their index admission (1992-1995). 290 patients met both Revised Diagnostic Interview for Borderlines and DSM-III-R criteria for borderline personality disorder and 72 met DSM-III-R criteria for at least 1 nonborderline Axis II disorder (and neither criteria set for borderline personality disorder). Over 94% of surviving patients were re-interviewed about their psychiatric treatment histories 2, 4, and 6 years later. RESULTS: Only 33% of borderline patients were hospitalized during the final 2 years of the 6-year follow-up, a substantial decline from the 79% who had prior hospitalizations at baseline. Much the same pattern emerged for day and/or residential treatment (from 55% to 22%). In contrast, about three quarters of borderline patients were still in psychotherapy and taking psychotropic medications after 6 years of follow-up. Additionally, over 70% of borderline patients participating in these outpatient modalities did so for at least 75% of each follow-up period. While rates of intensive psychotherapy declined significantly over time (from 36% to 16%), rates of intensive polypharmacy remained relatively stable over time, with about 40% of borderline patients taking 3 or more concurrent standing medications during each follow-up period, about 20% taking 4 or more, and about 10% taking 5 or more. CONCLUSIONS: The results of this study suggest that the majority of borderline patients continue to use outpatient treatment in a sustained manner through 6 years of follow-up, but only a declining minority use more restrictive and costly forms of treatment. PMID- 14744166 TI - A double-blind switch study of paroxetine and venlafaxine in obsessive-compulsive disorder. AB - BACKGROUND: The treatment guidelines for obsessive-compulsive disorder (OCD) propose to switch serotonin reuptake inhibitors (SRIs) in case of refractoriness. However, no controlled research has been published yet that prospectively examined the effects of changing SRIs. This article describes the first double blind switch study of 2 SRIs in patients with OCD. METHOD: 150 patients with primary OCD, according to DSM-IV criteria, were randomly assigned in a 12-week, double-blind trial to receive dosages titrated upward to 300 mg/day of venlafaxine (N = 75) or 60 mg/day of paroxetine (N = 75). Primary efficacy was assessed by the change from baseline on the Yale-Brown Obsessive-Compulsive Scale (Y-BOCS), and nonresponse was defined as less than 25% reduction on the Y-BOCS. After a 4-week tapering phase, 43 nonresponders were switched to 12 additional weeks of the alternate antidepressant, of which 16 patients received venlafaxine and 27 received paroxetine. RESULTS: Eighteen of 43 patients benefited from a switch to the alternate SRI with a mean +/- SD decrease of at least 25% on the Y BOCS. At the end of 12 weeks, responder rates were 56% for paroxetine (15/27) and 19% for venlafaxine (3/16). An intent-to-treat, last-observation-carried-forward analysis demonstrated a mean decrease on the Y-BOCS of 1.8 +/- 3.5 in the venlafaxine group and 6.5 +/- 7.1 in the paroxetine group. After 2 consecutive SRI trials, 109 of 150 patients (73%) achieved a Y-BOCS decrease of at least 25%. CONCLUSION: The results of the current study show that 42% of the nonresponders benefited from a crossover to the other SRI, and that paroxetine was more efficacious than venlafaxine in the treatment of nonresponders to a previous SRI trial. Switching SRIs in case of refractoriness may be considered a useful strategy for patients with OCD. PMID- 14744167 TI - Escitalopram continuation treatment prevents relapse of depressive episodes. AB - BACKGROUND: Current guidelines for antidepressant use recommend 4 to 6 months of continuation treatment to prevent relapse of depression following symptom resolution. This study evaluates the efficacy and safety of continuation escitalopram treatment. METHOD: Outpatients diagnosed with DSM-IV major depressive disorder (male or female, aged 18 to 81 years) who had completed 8 weeks of randomized double-blind treatment with escitalopram, citalopram, or placebo entered an 8-week flexible-dose, open-label phase in which all patients received escitalopram (10-20 mg/day). This study was initiated November 3, 1999, and completed April 5, 2001. Patients who met responder criteria (score of < or = 12 on the Montgomery-Asberg Depression Rating Scale [MADRS]) were randomly assigned in a 2:1 ratio to escitalopram (at the dose each patient was receiving at the end of the open-label phase) or placebo for 36 weeks of double-blind treatment. The primary efficacy variable was time to depression relapse (defined as MADRS score > or = 22 or discontinuation due to an insufficient therapeutic response) from the start of the double-blind treatment phase. RESULTS: A total of 502 patients received open-label escitalopram treatment and had at least 1 MADRS assessment. A total of 274 evaluable subjects entered the double-blind treatment phase; 93 received placebo and 181 received escitalopram. Time to depression relapse was significantly longer (p =.013) and the cumulative rate of relapse was significantly lower in patients who received escitalopram (26% escitalopram vs. 40% placebo; hazard ratio = 0.56; p =.01). Escitalopram-treated subjects had significantly lower depression ratings than those of placebo-treated patients. Escitalopram continuation treatment was safe and well tolerated. Discontinuation rates due to adverse events were 7% for the placebo group and 4% for the escitalopram-treated group. CONCLUSION: Continuation treatment with escitalopram is effective in preventing relapse into an episode of major depressive disorder. PMID- 14744168 TI - Zonisamide in the treatment of binge-eating disorder: an open-label, prospective trial. AB - BACKGROUND: Binge-eating disorder is characterized by recurrent episodes of uncontrollable overeating without compensatory weight-loss behaviors. It commonly co-occurs with obesity. Zonisamide is a novel antiepileptic drug associated with weight loss. The purpose of this study was to preliminarily assess zonisamide in the treatment of binge-eating disorder. METHOD: Fifteen outpatients with DSM-IV TR binge-eating disorder were enrolled from Jan. 25, 2002, through Sept. 10, 2002, in an open-label, prospective, 12-week, flexible dose (100-600 mg/day) study of zonisamide. The primary outcome measure was binge-eating episode frequency. Secondary measures included binge day frequency, body mass index (BMI), weight, Clinical Global Impressions-Severity of Illness scale (CGI-S) scores, Yale-Brown Obsessive-Compulsive Scale Modified for Binge Eating (YBOCS BE) scores, Three Factor Eating Questionnaire (TFEQ) scores, and Hamilton Rating Scale for Depression scores. Safety measures included adverse events, routine blood chemical and hematology laboratory values, urinalyses, plasma zonisamide levels, physical examination findings, and electrocardiograms. Outcome measures were analyzed by a repeated-measures random regression analysis using all data and an endpoint analysis using last observation carried forward. RESULTS: Eight subjects completed the 12 weeks of treatment. The mean (SD) zonisamide daily dose at endpoint evaluation was 513 (103) mg/day. Both the random regression and endpoint analyses found a highly significant decrease in binge-eating episode frequency, binge day frequency, BMI, weight, CGI-S scores, YBOCS-BE total scores, and TFEQ hunger and disinhibition scores (p <.0001 for all measures in both analyses except p =.001 for endpoint analysis of binge eating frequency, p =.0001 for endpoint analysis of TFEQ disinhibition, and p =.0008 for endpoint analysis of TFEQ hunger). The 7 subjects who discontinued zonisamide prematurely did so due to lack of response (N = 1), protocol non-adherence (N = 2), and adverse events (N = 4). CONCLUSION: Zonisamide was effective in reducing binge-eating frequency, severity of illness, and weight and was generally well tolerated in an open trial of binge-eating disorder. Controlled trials appear warranted. PMID- 14744169 TI - Prolactin levels in schizophrenia and schizoaffective disorder patients treated with clozapine, olanzapine, risperidone, or haloperidol. AB - BACKGROUND: Prolactin levels are elevated to varying degrees by antipsychotics. Prolactin elevations may result in sexual and other adverse effects, and they may be related to antipsychotic effects. We used the data collected in a trial of antipsychotics to study the differential effect of these drugs on prolactin level, to explore the relation between clinical effects and prolactin level, and to determine the relationship between plasma levels of antipsychotics and prolactin level. METHOD: Treatment-resistant patients (133 men, 24 women) diagnosed with DSM-IV schizophrenia or schizoaffective disorder participated in a double-blind, randomized, 14-week trial comparing clozapine (N = 40), olanzapine (N = 39), risperidone (N = 41), and haloperidol (N = 37). Plasma levels of prolactin and antipsychotics were determined at baseline and at weeks 5, 8, 10, 12, and 14 during the trial. Clinical effects were measured with the Positive and Negative Syndrome Scale and the Extrapyramidal Symptom Rating Scale. Statistical analyses were limited to the 75 men for whom repeated prolactin levels were available. Data were gathered from June 1996 to December 1999. RESULTS: Risperidone caused significant elevation of prolactin levels (p <.05) that appeared to be dose-dependent. Clozapine and olanzapine were associated with decreases of prolactin, whereas haloperidol led to a minor, nonsignificant increase. Plasma olanzapine and prolactin levels were correlated. Prolactin levels were not related to clinical improvement or extrapyramidal side effects. CONCLUSION: Antipsychotics show major differences in their effects on prolactin, and risperidone has clearly the most robust effect. PMID- 14744170 TI - A placebo-controlled trial of bupropion SR as an antidote for selective serotonin reuptake inhibitor-induced sexual dysfunction. AB - OBJECTIVE: This study reports the results of a placebo-controlled, double-blind comparison of bupropion sustained release (SR) as an antidote for sexual dysfunction versus placebo in 42 patients with selective serotonin reuptake inhibitor (SSRI)-induced sexual dysfunction. Exploratory analyses of the association of testosterone and sexual functioning in women in the study were also performed. METHOD: Patients with DSM-IV major depression who experienced a therapeutic response to any SSRI and were experiencing medication-induced global or phase-specific sexual dysfunction, as measured by the Changes in Sexual Functioning Questionnaire (CSFQ), were randomly assigned to receive either bupropion SR 150 mg b.i.d. or placebo for 4 weeks in addition to the SSRI. Total testosterone levels were assessed at baseline and week 4. RESULTS: The difference in global sexual functioning, based on the total CSFQ score, was not statistically significant between the 2 groups at week 4, nor were differences in orgasm, desire/ interest as measured by sexual thoughts, or self-reported arousal. There was a statistically significant difference between the 2 groups at week 4 in desire as measured by self-report feelings of desire and frequency of sexual activity. Desire/ frequency showed a significantly greater improvement among those patients receiving bupropion SR compared with placebo (Wilk's F = 5.47, df = 1, p =.024). Frequency was significantly correlated to total testosterone level at baseline (r = 0.36, p =.027) and at week 4 (r = 0.41, p =.025). CONCLUSIONS: Bupropion SR, as an effective antidote to SSRI-induced sexual dysfunction, produced an increase in desire to engage in sexual activity and frequency of engaging in sexual activity compared with placebo. A larger study is needed to further investigate this finding. PMID- 14744171 TI - Single-dose intravenous valproate in acute mania. AB - OBJECTIVE: High-dose loading with oral and intravenous valproate has been reported to be therapeutic in mania over 48 to 72 hours. We hypothesized that very high dose intravenous (IV) valproate might have even more rapid effects equivalent to effects in status epilepticus that occur within 20 minutes. METHOD: Seven patients with mania (Young Mania Rating Scale score > 20) and minimal prior drug treatment were given valproate 20 mg/kg IV over 30 minutes. RESULTS: No antimanic effects were observed during 120 minutes of observation. There were no side effects. CONCLUSION: Slowly evolving biochemical changes, perhaps at the gene level, may be required for the antimanic effect of anticonvulsants. PMID- 14744172 TI - In the puerperium, primiparae exhibit higher levels of anxiety and serum peptidase activity and greater immune responses than multiparae. AB - BACKGROUND: Delivery is accompanied by increases in anxiety levels that are significantly related to an activation of the inflammatory response system (IRS), as indicated by increases in the serum concentrations of interleukin-6 and the interleukin-1 receptor antagonist (IL-1RA) and increases in the activity of prolyl endopeptidase (PEP), a cytosolic endopeptidase that cleaves peptide bonds on the carboxyl side of proline in behaviorally active neuropeptides involved in anxiety. Primiparae may show an immune responsivity differing from that in multiparae. The aims of this study were to examine whether there are differences in anxiety levels, the IRS, and serum PEP values between primiparae and multiparae before and after delivery. METHOD: We administered the Spielberger State-Trait Anxiety Inventory (STAI) to and assessed serum IL-1RA and soluble CD8 (sCD8) concentrations and serum PEP activity in 48 primiparae and 48 multiparae at the end of term and 1 and 3 days after delivery. Data were gathered in 1996 and 1997. RESULTS: We found that in primiparae (p =.001), but not in multiparae (p =.6), there was a significant increase in the STAI score 3 days after delivery and that primiparae had significantly higher STAI scores than multiparae 3 days after delivery (p =.01). Primiparae showed significantly higher serum IL-1RA levels than multiparae 1 (p =.003) and 3 (p =.02) days after delivery, but not before delivery. Primiparae also had lower serum sCD8 and higher serum PEP activity than multiparae before and after delivery. CONCLUSIONS: The results suggest that primiparae suffer greater anxiety and have a different neuroimmune responsivity than multiparae and that the increased anxiety levels in primiparae may be related to changes in the IRS and serum PEP activity. PMID- 14744173 TI - Lifetime history of sexual abuse, clinical presentation, and outcome in a clinical trial for adolescent depression. AB - OBJECTIVE: To investigate the impact of sexual abuse on clinical presentation and treatment outcome in depressed adolescents. METHOD: 107 adolescent outpatients, 13 to 18 years old, with DSM-III-R major depression were randomly assigned to cognitive-behavioral therapy (CBT), systemic behavioral family therapy (SBFT), or nondirective supportive therapy (NST) from Oct. 1, 1991 through May 31, 1995. Subjects were classified on the basis of the presence or absence of lifetime history of sexual abuse. Since only 1 subject assigned to SBFT had a history of sexual abuse, we restricted our analyses to those 72 subjects assigned to either CBT or NST. The impact of lifetime history of sexual abuse on service use, depression, and treatment outcome was examined. RESULTS: Depressed adolescents with a past history of sexual abuse were more likely, at 2-year follow-up, to have had a psychiatric hospitalization and have a depressive relapse, even controlling for maternal depression, source of referral, race, and treatment assignment. CBT was more efficacious than NST in absence of sexual abuse but was not better than NST in those with a history of sexual abuse. CONCLUSION: Sexual abuse is a negative predictor of long-term outcome in adolescent depression. CBT for depression may not be as efficacious for those depressed adolescents with a history of sexual abuse. These findings suggest that a history of sexual abuse should be assessed not only in clinical practice, but also in research studies of depressive outcome. Further work is indicated to understand the relationship between sexual abuse and poor outcome in order to help restore these high-risk youths to an optimal developmental trajectory. PMID- 14744175 TI - One-year follow-up after successful ECT: a naturalistic study in depressed inpatients. AB - BACKGROUND: The aim of this study is to examine both long-term efficacy of electroconvulsive therapy (ECT) and the predictive value of adequate pre-ECT pharmacotherapy and the presence of delusions in relation to post-ECT relapse in patients who suffered from DSM-III-R major depression. METHOD: Forty responders (a decrease in Hamilton Rating Scale for Depression score > or = 50%) to ECT were followed for 1 year, the majority (N = 28) prospectively and the remainder (N = 12) retrospectively. Relapse was defined as readmission, an obvious decline in social functioning, or a change of antidepressant medication caused by a clear worsening of depressive symptoms. RESULTS: Both 6- and 12-month post-ECT relapse was significantly lower in patients with delusional depression compared with nondelusional patients: 3/24 (12%) versus 8/15 (53%) and 5/24 (21%) versus 11/15 (73%), respectively. Relapse rates for the whole sample were 11/39 (28%) at 6 months and 16/39 (41%) at 12 months. Regarding the impact of adequate pre-ECT antidepressant trials on relapse, our data are inconclusive, because only a few patients did not receive adequate pharmacotherapy prior to ECT. CONCLUSION: The remarkable finding of the present study is the favorable 1-year outcome for patients with delusional depression. The relapse rate for patients adequately pretreated with anti-depressants (45% over 1 year) is somewhat more favorable than expected. PMID- 14744174 TI - Gabapentin reduces cocaine use among addicts from a community clinic sample. AB - BACKGROUND: Individuals with chronic psychiatric conditions display a high rate of cocaine use. Gabapentin was hypothesized to reduce cocaine use by restoring inhibitory GABAergic feedback on ascending dopaminergic projections to nucleus accumbens neurons. METHOD: Nine participants with DSM-IV cocaine dependence were selected from patients attending a large community psychiatric clinic. During a 24-week open-label trial of gabapentin (800-2400 mg/day), qualitative urine drug screens were collected from the participants up to 3 times per week. Data were collected from September 1999 to May 2001. RESULTS: With gabapentin, the mean +/- SD number of cocaine-positive urine screens decreased from 53.11 +/- 13.23 to 35.22 +/- 14.84 (t = 3.58, N = 9, p <.01). The number of weeks of abstinence from cocaine increased from 2.1 +/- 1.5 to 8.0 +/- 5.5 (t = 3.21, N = 9, p <.01). CONCLUSION: Gabapentin appeared to be a safe and efficacious medication to reduce cocaine usage in a community sample of psychiatric patients. PMID- 14744176 TI - Branched chain amino acid treatment of tardive dyskinesia in children and adolescents. AB - BACKGROUND: A series of studies had demonstrated that deficient clearance of the large neutral amino acid phenylalanine was associated with tardive dyskinesia (TD), that the administration of the branched chain amino acids (BCAA) significantly decreased TD symptoms over placebo, and that the observed TD symptom reduction was significantly correlated with a diminished availability of phenylalanine to the brain of adult men with psychosis. As part of an initiative by the National Institute of Mental Health to expand the testing of treatments that were successful in adults to children and adolescents, the present pilot study was undertaken to test whether the BCAA would also reduce TD symptoms in children and adolescents. A 2-week trial of the BCAA was thus conducted in 6 children and adolescents (age range, 10.5-16.5 years) for the treatment of TD symptoms. METHOD: A clinical diagnosis of TD was made in all subjects on the basis of a global score derived from the Simpson Abbreviated Dyskinesia Rating Scale. Subjects were videotaped for TD evaluation at baseline and after 1 and 2 weeks of BCAA treatment given in the form of a drink administered 3 times daily. TD symptom change over the trial period was evaluated by researchers blinded to the treatment status of the evaluation. RESULTS: TD symptom decreases were substantial in 5 of the 6 participants, ranging from 40% to 65%. Two of the subjects received an additional course of treatment, and further reductions in TD symptoms over those seen in the 2-week trial were observed. CONCLUSION: The substantial symptom decrease and tolerability observed suggest the use of the BCAA formulation for the treatment of TD in children and adolescents and warrant further large-scale studies. PMID- 14744177 TI - An open-label trial of sildenafil addition in risperidone-treated male schizophrenia patients with erectile dysfunction. AB - BACKGROUND: Sexual dysfunction frequently occurs in treated and untreated patients with schizophrenia. Sildenafil is used for treatment of erectile dysfunction caused by diverse factors. The aim of our study was to evaluate its potential value, safety, and effect on compliance with anti-psychotic medications in risperidone-treated male schizophrenia patients suffering from erectile dysfunction. METHOD: In a 6-week open-label trial, sildenafil was administered to 12 male schizophrenia (DSM-IV) patients, treated with risperidone and reporting erectile dysfunction. The starting dose was 25 mg with the possibility to increase the dose to 75 mg. Three patients who did not respond stopped sildenafil treatment after 3 weeks. The effect on sexual function was assessed by the International Index of Erectile Function and the Valevski-Weizman Male Sexual Function scale. RESULTS: Nine (75%) of the 12 patients completed the 6-week trial, and 3 patients (25%) stopped taking sildenafil after 3 weeks due to lack of response. We observed statistically significant improvements in all sexual function domains (desire, erectile function, orgasmic function, intercourse satisfaction, and overall satisfaction) in the 9 patients who completed the trial and in most of the domains for all 12 study participants. More than half (8/12; 67%) of the patients exhibited partial or much improvement. CONCLUSION: Sildenafil is a useful agent for the treatment of erectile dysfunction in risperidone-treated male schizophrenia patients. PMID- 14744178 TI - Olanzapine versus placebo in the treatment of borderline personality disorder. AB - BACKGROUND: Atypical antipsychotics are increasingly used in clinical practice in the management of borderline personality disorder (BPD), and a small but growing body of literature supports their efficacy. Here, we report the results of a double-blind, placebo-controlled study of olanzapine as a treatment for BPD. METHOD: Forty BPD patients (25 female, 15 male) were randomly assigned in equal numbers to olanzapine and placebo. Diagnoses were made using the Structured Clinical Interview for DSM-IV Axis II Personality Disorders and the Mini International Neuropsychiatric Interview. Patients with schizophrenia, bipolar disorder, or current major depression were excluded. Olanzapine dosage was flexible, and the dose range was 2.5 to 20 mg/day, with most patients receiving 5 to 10 mg/day. No concomitant psychotropic medications were allowed. Patients were assessed at baseline and at 2, 4, 8, and 12 weeks. The primary outcome was change in the total score for the 9 BPD criteria on a 1-to-7 Likert scale, the Clinical Global Impressions scale modified for borderline personality disorder (CGI-BPD), using an analysis of covariance model including baseline score as covariate. Data were collected from July 2000 to April 2002. RESULTS: Olanzapine was found to be significantly (p <.05) superior to placebo on the CGI-BPD at endpoint, with separation occurring as early as 4 weeks. Similar results were found for the single-item Clinical Global Impressions scale. Weight gain was significantly (p =.027) greater in the olanzapine group. CONCLUSIONS: This study supports the efficacy of olanzapine for symptoms of BPD in a mixed sample of women and men. Further studies with olanzapine and other atypical antipsychotics are needed. PMID- 14744179 TI - The effect of a switch to ziprasidone in an adult population with autistic disorder: chart review of naturalistic, open-label treatment. AB - BACKGROUND: The present investigation retrospectively assessed the effect of an open-label switch to ziprasidone from other atypical antipsychotics on behavior, weight, and lipid levels in an adult population with autistic disorder. METHOD: We conducted a chart review of 10 adults (mean +/- SD age = 43.8 +/- 6.0 years) with DSM-IV autistic disorder who were switched from other atypical antipsychotics to ziprasidone, primarily due to weight gain, but other reasons included hypercholesterolemia, maladaptive behaviors, drowsiness, and depression. They had been treated with ziprasidone for at least 6 months. Our review focused on frequency of maladaptive behaviors, weight, and lipid levels. RESULTS: The mean +/- SD daily dose of ziprasidone was 128 +/- 41 mg, and all 10 patients continued with this same treatment after completion of the 6-month trial. Seven patients were found to have an improvement or no change in their maladaptive behavior. Eight patients (80%) lost weight (mean change = -13.1 +/- 7.0 lb [-5.9 +/- 3.2 kg]), 4 (80%) of 5 patients had a decrease in total cholesterol level, and 3 (60%) of 5 had a decrease in triglyceride levels. Data on lipid levels were available for 5 of the 10 patients. Behavioral activation was not noted in this population. There were no significant adverse effects associated with ziprasidone. CONCLUSION: In adults with autism, a switch to ziprasidone from other atypical antipsychotics appears to have the potential for maintaining beneficial effect on behavior while improving major health indices including weight and lipid levels. PMID- 14744180 TI - A 3-month, randomized, placebo-controlled, neuroleptic discontinuation study in 100 people with dementia: the neuropsychiatric inventory median cutoff is a predictor of clinical outcome. AB - BACKGROUND: Although few placebo-controlled neuroleptic discontinuation studies have been conducted in people with dementia, such studies are essential to inform key clinical decisions. METHOD: A 3-month, double-blind, placebo-controlled, neuroleptic discontinuation study (June 2000 to June 2002) was completed in 100 care-facility residents with probable or possible Alzheimer's disease (according to National Institute of Neurological and Communicative Diseases and Stroke/Alzheimer's Disease and Related Disorders Association criteria) who had no severe behavioral disturbances and had been taking neuroleptics for longer than 3 months. The Neuropsychiatric Inventory (NPI) was used to measure changes in behavioral and psychiatric symptoms. Quality of life was evaluated using Dementia Care Mapping. RESULTS: Eighty-two patients completed the 1-month assessment (36 placebo, 46 active). The number of participants withdrawing overall (N = 14 [30%] placebo, N = 14 [26%] active treatment) and because of exacerbation of behavioral symptoms (N = 6 [13%] placebo, N = 5 [9%] active treatment) was similar in the neuroleptic- and placebo-treated patients. As hypothesized, patients with baseline NPI scores at or below the median (< or = 14) had a particularly good outcome, with a significantly greater reduction of agitation in the patients receiving placebo (Mann-Whitney U test, z = 2.4, p =.018), while patients with higher baseline NPI scores were significantly more likely to develop marked behavioral problems if discontinued from neuroleptics (chi(2) = 6.8, p =.009). There was no overall difference in the change of quality of life parameters between groups. DISCUSSION: A standardized evaluation with an instrument such as the NPI may be a clinical indicator of which people with dementia are likely to benefit from discontinuation of neuroleptic treatment. PMID- 14744181 TI - Academic highlights: guidelines for the use of long-acting injectable atypical antipsychotics. PMID- 14744182 TI - Hypomanic episodes after receiving ziprasidone: an unintended "on-off-on" course of treatment. PMID- 14744183 TI - Intelligence, memory, and handedness in pedophilia. AB - A sample of 473 male patients with pedophilia (assessed by the patients' sexual history and penile response in the laboratory to standardized, erotic stimuli) or other problematic sexual interests or behaviors received brief neuropsychological assessments. Neuropsychological measures included a short form of the Wechsler Adult Intelligence Scale--Revised (D. Wechsler, 1981), the Hopkins Verbal Learning Test--Revised (R. H. B. Benedict, D. Schretlen. L. Groninger. & J. Brandt, 1998), the Brief Visuospatial Memory Test--Revised (R. H. B. Benedict, 1997), and the Edinburgh Handedness Inventory (S. M. Williams, 1986). Pedophilia showed significant negative correlations with IQ and immediate and delayed recall memory. Pedophilia was also related to non-right-handedness even after covarying age and IQ. These results suggest that pedophilia is linked to early neurodevelopmental perturbations. PMID- 14744184 TI - Memory for visuospatial location following selective hippocampal sclerosis: the use of different coordinate systems. AB - This study addressed the role of the medial temporal lobe regions and, more specifically, the contribution of the human hippocampus in memory for body centered (egocentric) and environment-centered (allocentric) spatial location. Twenty-one patients with unilateral atrophy of the hippocampus secondary to long standing epilepsy (left, n = 7; right, n = 14) and 15 normal control participants underwent 3 tasks measuring recall of egocentric or allocentric spatial location. Patients with left hippocampal sclerosis were consistently impaired in the allocentric conditions of all 3 tasks but not in the egocentric conditions. Patients with right hippocampal sclerosis were impaired to a lesser extent and in only 2 of the 3 tasks. It was concluded that hippocampal structures are crucial for allocentric, but not egocentric, spatial memory. PMID- 14744185 TI - Reading and phonological awareness in Williams syndrome. AB - This article describes the relationship between reading, phonological awareness abilities (PA), and intelligence in a group of 16 individuals with Williams syndrome (WS) and in a group of 16 typically developing children, matched for mental age. The individuals with WS were impaired in passage comprehension, in some areas of PA investigated (syllable deletion and rhyme detection), and in nonword reading accuracy, a measure of grapheme-phoneme conversion. This latter finding is relevant, considering that in Italy regular print-to-sound correspondence is the most practiced teaching routine in the early phases of learning to read. PMID- 14744186 TI - Use of IQ-adjusted norms to predict progressive cognitive decline in highly intelligent older individuals. AB - Identifying high-functioning older individuals in preclinical phases of Alzheimer's disease (AD) may require more sensitive methods than the standard approach. The authors explored the utility of adjusting for premorbid intelligence to predict progressive cognitive decline or Mild Cognitive Impairment (MCI) in 42 highly intelligent older individuals. When scores were adjusted for baseline IQ, 9 participants had executive impairments, 11 had memory impairments, and 22 scored in the normal range. None were impaired according to standard age norms. Three and a half years later, 9 participants with IQ-adjusted memory impairment declined in naming, visuospatial functioning, and memory; 6 convened to MCI. Three participants with normal memory declined. Implications for using IQ-adjusted norms to predict preclinical AD are discussed. PMID- 14744187 TI - Stroop tasks reveal abnormal selective attention among psychopathic offenders. AB - Selective attention among offenders with psychopathy was investigated using 3 Stroop paradigms: a standard color-word (CW) Stroop, a picture-word (PW) Stroop, and a color-word Stroop in which the word and color were spatially separated (separated CW). Consistent with "overselective" attention, offenders with psychopathy displayed reduced Stroop interference on the separated CW and PW tasks relative to offenders who were not psychopathic. However, offenders with psychopathy displayed normal Stroop interference on the standard CW Stroop. Further, the reduced interference of offenders with psychopathy on the separated CW Stroop was accompanied by normal facilitation. These findings suggest a circumscribed attentional deficit in psychopathy that hinders the use of unattended information that is (a) not integrated with deliberately attended information and (b) not compatible with current goal-directed behavior. PMID- 14744188 TI - Confirmatory factor analysis of the california verbal learning test in patients with epilepsy: relationship to clinical and neuropathological markers of temporal lobe epilepsy. AB - Latent constructs involved in California Verbal Learning Test (D. C. Delis, J. H. Kramer, E. Kaplan, & B. A. Ober, 1987) performance were examined using confirmatory factor analysis in 388 epilepsy surgery candidates. Eight factor models were compared. A single-factor model was examined, along with 7 models accommodating constructs of auditory attention, inaccurate recall, and delayed recall in different combinations. The retained model consisted of 3 correlated factors: Auditory Attention. Verbal Learning, and Inaccurate Recall. Validity of this factor structure was examined in a subsample of patients with left and right temporal lobe epilepsy. All 3 factors were related to seizure focus and magnetic resonance imaging hippocampal volume. Only Verbal Learning was related to hippocampal neuropathology, supporting the distinction between learning and attention in the factor structure. PMID- 14744189 TI - Working memory and perseveration in verbal fluency. AB - Letter and semantic fluency tasks are often used in neuropsychological assessment and are sensitive to many conditions. Performance is assessed by correct responses and errors, including perseverations. Healthy young adults performed letter and semantic fluency tasks. One group performed these tasks in the conventional manner; 2 other groups performed them while maintaining memory loads. The memory loads consisted either of words from the same category as the fluency task or of words from a different category. The results showed little effect of memory loads on correct responses but significant effects of memory load on perseveration rates: Same-category loads resulted in higher rates, especially in letter fluency. The results are discussed in terms of frontal lobe function in verbal fluency. PMID- 14744190 TI - Contribution of organizational strategy to verbal learning and memory in adults with attention-deficit/hyperactivity disorder. AB - Statistical mediation modeling was used to test the hypothesis that poor use of a semantic organizational strategy contributes to verbal learning and memory deficits in adults with attention-deficit/hyperactivity disorder (ADHD). Comparison of 28 adults with ADHD and 34 healthy controls revealed lower performance by the ADHD group on tests of verbal learning and memory, sustained attention, and use of semantic organization during encoding. Mediation modeling indicated that state anxiety, but not semantic organization, significantly contributed to the prediction of both learning and delayed recall in the ADHD group. The pattern of findings suggests that decreased verbal learning and memory in adult ADHD is due in part to situational anxiety and not to poor use of organizational strategies during encoding. PMID- 14744191 TI - The relationship between neuropsychological functioning and driving ability in dementia: a meta-analysis. AB - A meta-analysis of 27 primary studies was conducted to examine the relationship between neuropsychological functioning and driving ability for adults with dementia. When studies using a control group were included, the relationship between cognitive measures and on-road or non-road driving measures was significant for all reported domains; mean correlations ranged from.35 to.65. Caregiver reports of driving ability and cognitive variables were correlated significantly only on measures of mental status and visuospatial skills. When studies using a control group were excluded, moderate mean correlations were observed for visuospatial skills and on-road or non-road measures, and for mental status with non-road tests. Other effects were small or nonsignificant. Implications for basing driving recommendations on neuropsychological testing are discussed. PMID- 14744192 TI - Elevated false recognition in patients with frontal lobe damage is neither a general nor a unitary phenomenon. AB - This study examined verbal recognition memory in amnesic patients with frontal lesions (AF), nonamnesic patients with frontal lesions (NAF), and amnesic patients with medial temporal lesions (MT). To examine susceptibility to false alarms, the number of studied words drawn from various categories was varied. The AF and MT groups demonstrated reduced hits and increased false alarms. False alarms were especially elevated when item-specific recollection was strongest in control participants. The NAF group performed indistinguishably from control participants, but several patients showed excessive false alarms in the context of normal hit rates. These patients exhibited impaired monitoring and verification processes. The findings demonstrate that elevated false recognition is not characteristic of all frontal patients and may result from more than 1 underlying mechanism. PMID- 14744193 TI - Retrieval inhibition in directed forgetting following severe closed-head injury. AB - A variant of the list method directed forgetting procedure was used to examine the role of inhibition in memory performance following severe closed-head injury (CHI). Twenty-four participants with severe CHI and 24 controls studied picture and word stimuli in both forget and remember conditions. Memory testing for the to-be-forgotten and to-be-remembered items consisted of a free-recall test followed by a source-monitoring task. Despite poorer recall performance, the participants with CHI exhibited a directed forgetting effect similar to that in controls. Item recognition scores indicated that the inhibited items were not forgotten but rather were items whose accessibility had been lowered. These findings suggest that residual memory deficits in patients with severe CHI are unlikely to reflect inefficient retrieval inhibition. PMID- 14744194 TI - Control of fixation and saccades in humans with chronic lesions of oculomotor cortex. AB - To elucidate the dynamic interactions of cortical and subcortical oculomotor systems, the authors investigated reflexive and strategic control over fixation and saccades in patients with chronic unilateral lesions that involved either frontal or parietal cortex. They measured the effects of indicating the location of the forthcoming target and removing the fixation stimulus on the latencies of eye movements toward a peripheral visual target in 12 patients with frontal eye field (FEF) lesions, 9 patients with lesions restricted to parietal cortex, and 12 neurologically healthy controls. They found that chronic damage to FEF cortex disrupts cortico-collicular interactions, resulting in hypoactivity in the contralesional superior colliculus and a loss of strategic control over the intrinsic collicular circuits that regulate fixation. PMID- 14744196 TI - A lexical stress effect in neglect dyslexia. AB - The aim of this study was to investigate the effect of word stress location in neglect dyslexia. An assessment was made of stress assignment in reading Italian words in 13 patients with neglect dyslexia and 13 matched normal individuals. The correspondence in stress location between target and neglect errors was preserved in 9 patients. Moreover, the proportion of words with stress on the penultimate syllable was about 50% and that on the antepenultimate syllable was also about 50%. This pattern is significantly different from the distribution observed in the Italian lexicon, in which the majority of polysyllabic words are stressed on the penultimate syllable. This result cannot be expected from the application of a "regularization" strategy associated with the operation of a nonlexical reading pathway. Taken together, these findings lead to the conclusion that lexical stress is part of the lexical information preserved in neglect dyslexia. PMID- 14744195 TI - Implicit spatial contextual learning in healthy aging. AB - Three experiments investigated the aging of implicit spatial and spatiotemporal context learning in 2 tasks. In contextual cuing, people learn to use repeated spatial configurations to facilitate search for a target, whereas in higher order serial learning, they learn to use subtle sequence regularities to respond more quickly and accurately to a series of events. Results reveal a dissociation; overall contextual cuing is spared in healthy aging, whereas higher order sequence learning is impaired in the same individuals. This finding suggests that these 2 forms of implicit learning rely on different neural substrates that age differently; the results are also consistent with recent evidence that fronto striatal circuits are particularly susceptible to decline in health aging. PMID- 14744197 TI - Lateralization of epileptic foci by neuropsychological testing in mesial temporal lobe epilepsy. AB - This study investigated the lateralizing value of neuropsychological testing in presurgical evaluation of mesial temporal lobe epilepsy (MTLE). This study differed from previous ones in that the cutoff values were determined to yield high positive predictive values (PPVs), multiple neuropsychological predictors were considered in combination, and patients with atypical language dominance or low intelligence were not excluded from the sample. The participants were 92 patients with MTLE (left, n = 47; right, n = 45) who showed good postoperative seizure control. With a stringent cutoff criterion, the multiple neuropsychological predictors considered in combination yielded a sensitivity of 15% and a PPV of 93%, and with a less stringent cutoff criterion, a sensitivity of 37% and a PPV of 83%. PMID- 14744198 TI - Impaired decision making related to working memory deficits in individuals with substance addictions. AB - This study examined whether individuals with substance dependence (ISDs) show impairments in working memory and whether there is a relationship between their impairments in decision making as measured by the gambling task (GT) paradigm and working memory as measured by a delayed nonmatching to sample (DNMS) task. Using the GT, 11% of healthy control participants and 61% of ISDs opted for choices with high immediate gains in spite of higher future losses. For the ISDs and controls with equal GT impairments, the ISDs performed significantly lower than controls on the DNMS task. The nonimpaired ISDs on the GT also performed significantly worse than matched controls on the DNMS task. The DNMS task deficit in ISDs was across all delay times, suggesting the deficit may lie in the "executive" process of working memory, which supports earlier findings (E. M. Martin et al., 2003). The authors suggest that the prefrontal cortex hosts multiple distinct mechanisms of decision making and inhibitory control and that ISDs may be affected in any one or combination of them. PMID- 14744199 TI - Selection ability in Alzheimer's disease: investigation of a component of semantic processing. AB - Selection ability (selecting a response from several competing semantic and/or lexical representations) was tested in 21 participants with Alzheimer's disease (AD) and 28 control participants to help clarify the nature of semantic impairments in AD. Selection demands were manipulated in 3 tasks (lexical fluency, comparison, and verb generation). In each, high-selection conditions required response selection from competing alternatives, whereas low-selection conditions had a reduced need for selection. Patients with AD were disproportionately impaired on the high-selection conditions of all tasks, even when this condition was easier. Selection deficits on verb generation were evident only relative to nonspeeded controls. Overall results indicate impaired semantic selection abilities in AD, which may contribute to poor performance on some semantic tasks. PMID- 14744200 TI - Orthography and the hemispheres: visual and linguistic aspects of letter processing. AB - Hebrew and Arabic are Semitic languages with a similar morphological structure and orthographies that differ in visual complexity. Two experiments explored the interaction of the characteristics of orthography and hemispheric abilities on lateralized versions of a letter-matching task (Experiment 1) and a global-local task (Experiment 2). In Experiment 1, native Hebrew readers and native Arabic readers fluent in Hebrew matched letters in the 2 orthographies. The results support the hypothesis that Arabic orthography is more difficult than Hebrew orthography for participants who can read both languages and that this difficulty has its strongest effects in the left visual field. In Experiment 2, native Arabic speakers performed a global-local letter detection task with Arabic letters with 2 types of inconsistent stimuli: different and similar. The results support the hypothesis that the right hemisphere of skilled Arabic readers cannot distinguish between similar Arabic letters, whereas the left hemisphere can. PMID- 14744202 TI - Case reports: slices of real life to complement evidence. PMID- 14744201 TI - Effects of perinatal exposure to PCBs on neuropsychological functions in the Rotterdam cohort at 9 years of age. AB - PCBs are known for their neurotoxic properties, especially on the developing brain. To increase insight into the neurotoxic effects of PCB exposure, the authors studied the effects of perinatal exposure to environmental levels of these compounds on different neuropsychological domains. In 9-year-old children of the Rotterdam PCB--dioxin cohort, higher prenatal PCB levels were associated with longer response times (RTs), more variation in RTs, and lower scores on the Tower of London (TOL; Shallice, 1982). A longer breast-feeding duration was associated with lower TOL scores and with better spatial organizational skills. There was some evidence of negative effects of lactational exposure to PCBs on scores on the TOL. PMID- 14744203 TI - Relationships between standing balance and symmetry measurements in patients following recent strokes (3 weeks or less) or older strokes (6 months or more). AB - BACKGROUND AND PURPOSE: The Functional Standing Balance (FSB) Scale was designed to obtain measurements of standing balance and to identify the problems typically faced by people with stroke. The purpose of this study was to investigate the validity of measurements obtained with the FSB Scale for use in the acute and chronic phases of stroke by comparing the measurements obtained with the FSB Scale with those obtained for postural sway and lateral symmetry by use of a force platform. SUBJECTS AND METHODS: Measurements were obtained for 26 people with recent strokes (ie, strokes within 3 weeks of data collection) and for 28 people with long-standing strokes (ie, strokes of 6 months' duration or older). The FSB Scale consists of 3 components: weight distribution, balance without movement, and balance with movement. Measurements of balance performance were compared with measurements of anteroposterior and lateral sway velocity obtained on a force platform. The weight distribution on 2 digital scales was compared with the lateral symmetry measured on the force platform. RESULTS: The highest correlations were found between the FSB Scale balance measurements and the measurements of anteroposterior sway velocity obtained on the force platform with feet apart and eyes open. The correlations (r) were -.68 and -.67 for the group with recent strokes and -.74 and -.91 for the group with long-standing strokes. The correlations (r) between weight distribution measured on the digital scales and lateral symmetry measured on the force platform were.44 for the group with recent strokes and.52 for the group with long-standing strokes. DISCUSSION AND CONCLUSION: The subjects whose results on the FSB Scale were poor had higher sway velocities on the force platform than the subjects whose results on the FSB Scale were good. The results of this study suggest that the FSB Scale provides the same kind of information as that obtained for sway velocity and lateral symmetry as measured with the use of force platforms in both patients with recent strokes and patients with long-standing strokes. PMID- 14744204 TI - Leadership, administration, management, and professionalism (LAMP) in physical therapy: a Delphi study. AB - BACKGROUND AND PURPOSE: The knowledge and skills needed by physical therapists entering practice in the areas of leadership, administration, management, and professionalism (LAMP) are not known. Using the LAMP components identified by American Physical Therapy Association's Section on Administration, this study sought to define the range of LAMP content pertinent to physical therapy clinical management and to explore LAMP knowledge and skills required of physical therapists upon entry into the profession. SUBJECTS AND METHODS: Thirty-four physical therapist managers participated in a Delphi study to (1) create a comprehensive list of defined LAMP components, (2) determine the perceived importance of each component in the management of clinical practices, and (3) identify the level of knowledge and skill for each component believed to be necessary for a new physical therapist graduate. RESULTS: Respondents agreed that 178 items should be on the LAMP component list. They perceived that almost all LAMP components are important in the management of a clinical practice, and they indicated that new graduates needed moderate to extensive knowledge in 44% of them. They believed that new graduates needed no skill in 29% of the components, whereas they needed at least intermediate skill for 22% of them. Top-ranked component categories across the 3 scales (importance, knowledge, and skill) were communication, professional involvement and ethical practice, delegation and supervision, stress management, reimbursement sources, time management, and health care industry scanning. DISCUSSION AND CONCLUSION: This study provides a basis for further exploration of which LAMP components should be included in professional (entry-level) physical therapist curricula and which components should be learned after graduation. PMID- 14744205 TI - Optimizing the sensitivity of the head thrust test for identifying vestibular hypofunction. AB - BACKGROUND AND PURPOSE: The head thrust test (HTT) is used to assess the vestibulo-ocular reflex. Sensitivity and specificity for diagnosing unilateral vestibular hypofunction (UVH) in patients following vestibular ablation is excellent (100%), although sensitivity is lower (35%-39%) for patients with nonsurgically induced UVH. The variability of the test results may be from moving the subject's head outside the plane of the lateral semicircular canals as well as using a head thrust of predictable timing and direction. The purpose of this study was to examine sensitivity and specificity of the horizontal HTT in identifying patients with UVH and bilateral vestibular hypofunction (BVH) when the head was flexed 30 degrees in attempt to induce acceleration primarily in the lateral semicircular canal and the head was moved unpredictably. SUBJECTS: The medical records of 176 people with and without vestibular dysfunction (n=79 with UVH, n=32 with BVH, and n=65 with nonvestibular dizziness) were studied. METHODS: Data were retrospectively tabulated from a de-identified database (ie, with health information stripped of all identifiers). RESULTS: Sensitivity of the HTT for identifying vestibular hypofunction was 71% for UVH and 84% for BVH. Specificity was 82%. DISCUSSION AND CONCLUSION: Ensuring the head is pitched 30 degrees down and thrust with an unpredictable timing and direction appears to improve sensitivity of the HTT. PMID- 14744206 TI - Relationship between spontaneous kicking and age of walking attainment in preterm infants with very low birth weight and full-term infants. AB - BACKGROUND AND PURPOSES: Although early kicking differences have been reported for preterm infants without overt cranial sonographic abnormalities, their functional importance remains unclear because no outcomes have been measured. Therefore, the first purpose of this prospective study was to examine the age of walking attainment in preterm infants who had very low birth weight (VLBW) but no overt neurosonographic abnormalities and full-term infants without known impairments or pathology. The second purpose was to examine the relationship between spontaneous kicking and age of walking attainment in these infants. SUBJECTS AND METHODS: Twenty-two preterm infants and 22 full-term infants were examined for kicking movements at 2 and 4 months corrected age and were followed up for age of walking attainment until 18 months corrected age. RESULTS: Survival analysis showed that infants with VLBW attained walking ability at older ages than full-term infants after correction for prematurity. Cox proportional-hazards regression analyses for all infants revealed that a high hip-knee correlation at 2 months corrected age, a high kick frequency at 4 months corrected age, and a short intra-kick pause together with a low variability in interlimb coordination at 2 and 4 months corrected age were all associated with a decreased rate of walking attainment. DISCUSSION AND CONCLUSION: The results indicated that preterm infants who had VLBW but no overt neurosonographic abnormalities had an increased risk of delayed walking attainment compared with full-term infants. Alterations of spontaneous kicking may predict a decreased rate of walking attainment in both preterm and full-term infants. PMID- 14744208 TI - Toward an integrative science of the person. AB - To build a science of the person, the most basic question was, and remains, how can one identify and understand the psychological invariance that distinctively characterizes an individual and that underlies the variations in the thoughts, feelings, and actions that occur across contexts and over time? This question proved particularly difficult because of the discrepancies that soon emerged between the expressions of consistency that were expected and those that were found. The resulting dilemma became known as the classic "personality paradox": How can we reconcile our intuitions-and theories-about the invariance and stability of personality with the equally compelling empirical evidence for the variability of the person's behavior across diverse situations? Which is right: the intuitions or the research findings? In this chapter I review and discuss some of the advances made to answer this question since it was posed. These findings have allowed a resolution of the paradox, and provide the outlines for a conception of the underlying structure and dynamics of personality that seems to better account for the data. PMID- 14744207 TI - Factors related to the inability of individuals with low back pain to improve with a spinal manipulation. AB - BACKGROUND AND PURPOSE: Although spinal manipulation is one of the few interventions for low back pain supported by evidence, it appears to be underutilized by physical therapists, possibly due to therapists' concerns that a patient may not benefit from the intervention. The purpose of this study was to identify factors that are associated with an inability to benefit from manipulation. SUBJECTS: Seventy-five people with nonradicular low back pain (mean age=37.6 years, SD=10.6, range=19-59; mean duration of symptoms=41.7 days, SD=54.7, range=1-252) participated. METHODS: Subjects underwent a standardized examination that included history-taking; self-reports of pain, disability, and fear-avoidance beliefs; measurement of lumbar and hip range of motion; and use of various tests. All subjects received a spinal manipulation intervention for a maximum of 2 sessions. Subjects who did not show greater than 5 points of improvement on the modified Oswestry Low Back Pain Disability Questionnaire were considered to have shown no improvement with the manipulation. Baseline variables were tested for univariate relationship with the outcome of the manipulation. Variables showing a univariate relationship were entered into a logistic regression equation, and adjusted odds ratios were calculated. RESULTS: Twenty subjects (28%) did not improve with manipulation. Six variables were identified as being related to inability to improve with manipulation: longer symptom duration, having symptoms in the buttock or leg, absence of lumbar hypomobility, less hip rotation range of motion, less discrepancy in left-to-right hip medial rotation range of motion, and a negative Gaenslen sign. The resulting logistic regression model explained 63% of the variance in manipulation outcome. DISCUSSION AND CONCLUSION: The majority of subjects improved with manipulation. Baseline variables could be identified that were predictive of which subjects would not improve. PMID- 14744209 TI - On building a bridge between brain and behavior. AB - Cognitive neuroscience is motivated by the precept that a discoverable correspondence exists between mental states and brain states. This precept seems to be supported by remarkable observations and conclusions derived from event related potentials and functional imaging with humans and neurophysiology with behaving monkeys. This review evaluates specific conceptual and technical limits of claims of correspondence between neural events, overt behavior, and hypothesized covert processes examined using data on the neural control of saccadic eye movements. PMID- 14744210 TI - The neurobiology of consolidations, or, how stable is the engram? AB - Consolidation is the progressive postacquisition stabilization of long-term memory. The term is commonly used to refer to two types of processes: synaptic consolidation, which is accomplished within the first minutes to hours after learning and occurs in all memory systems studied so far; and system consolidation, which takes much longer, and in which memories that are initially dependent upon the hippocampus undergo reorganization and may become hippocampal independent. The textbook account of consolidation is that for any item in memory, consolidation starts and ends just once. Recently, a heated debate has been revitalized on whether this is indeed the case, or, alternatively, whether memories become labile and must undergo some form of renewed consolidation every time they are activated. This debate focuses attention on fundamental issues concerning the nature of the memory trace, its maturation, persistence, retrievability, and modifiability. PMID- 14744211 TI - Understanding other minds: linking developmental psychology and functional neuroimaging. AB - Evidence from developmental psychology suggests that understanding other minds constitutes a special domain of cognition with at least two components: an early developing system for reasoning about goals, perceptions, and emotions, and a later-developing system for representing the contents of beliefs. Neuroimaging reinforces and elaborates upon this view by providing evidence that (a) domain specific brain regions exist for representing belief contents, (b) these regions are apparently distinct from other regions engaged in reasoning about goals and actions (suggesting that the two developmental stages reflect the emergence of two distinct systems, rather than the elaboration of a single system), and (c) these regions are distinct from brain regions engaged in inhibitory control and in syntactic processing. The clear neural distinction between these processes is evidence that belief attribution is not dependent on either inhibitory control or syntax, but is subserved by a specialized neural system for theory of mind. PMID- 14744212 TI - Hypocretin (orexin): role in normal behavior and neuropathology. AB - The hypocretins (Hcrts, also known as orexins) are two peptides, both synthesized by a small group of neurons, most of which are in the lateral hypothalamic and perifornical regions of the hypothalamus. The hypothalamic Hcrt system directly and strongly innervates and potently excites noradrenergic, dopaminergic, serotonergic, histaminergic, and cholinergic neurons. Hcrt also has a major role in modulating the release of glutamate and other amino acid transmitters. Behavioral investigations have revealed that Hcrt is released at high levels in active waking and rapid eye movement (REM) sleep and at minimal levels in non-REM sleep. Hcrt release in waking is increased markedly during periods of increased motor activity relative to levels in quiet, alert waking. Evidence for a role for Hcrt in food intake regulation is inconsistent. I hypothesize that Hcrt's major role is to facilitate motor activity tonically and phasically in association with motivated behaviors and to coordinate this facilitation with the activation of attentional and sensory systems. Degeneration of Hcrt neurons or genetic mutations that prevent the normal synthesis of Hcrt or of its receptors causes human and animal narcolepsy. Narcolepsy is characterized by an impaired ability to maintain alertness for long periods and by sudden losses of muscle tone (cataplexy). Administration of Hcrt can reverse symptoms of narcolepsy in animals, may be effective in treating human narcolepsy, and may affect a broad range of motivated behaviors. PMID- 14744213 TI - Speech perception. AB - This chapter focuses on one of the first steps in comprehending spoken language: How do listeners extract the most fundamental linguistic elements-consonants and vowels, or the distinctive features which compose them-from the acoustic signal? We begin by describing three major theoretical perspectives on the perception of speech. Then we review several lines of research that are relevant to distinguishing these perspectives. The research topics surveyed include categorical perception, phonetic context effects, learning of speech and related nonspeech categories, and the relation between speech perception and production. Finally, we describe challenges facing each of the major theoretical perspectives on speech perception. PMID- 14744214 TI - Visual mechanisms of motion analysis and motion perception. AB - Psychophysical experiments on feature tracking suggest that most of our sensitivity to chromatic motion and to second-order motion depends on feature tracking. There is no reason to suppose that the visual system contains motion sensors dedicated to the analysis of second-order motion. Current psychophysical and physiological data indicate that local motion sensors are selective for orientation and spatial frequency but they do not eliminate any of the three main models-the Reichardt detector, the motion-energy filter, and gradient-based sensors. Both psychophysical and physiological data suggest that both broadly oriented and narrowly oriented motion sensors are important in the early analysis of motion in two dimensions. PMID- 14744215 TI - Cumulative progress in formal theories of attention. AB - Formal theories of attention based on similarity-choice theory and signal detection theory are reviewed to document cumulative progress in theoretical understanding of attention from the 1950s to the present. Theories based on these models have been developed to account for a wide variety of attentional phenomena, including attention to dimensions, attention to objects, and executive control. The review describes the classical similarity-choice and signal detection theories and relates them to current theories of categorization, Garner tasks, visual search, cuing procedures, task switching, and strategy choice. PMID- 14744216 TI - The psychology and neuroscience of forgetting. AB - Traditional theories of forgetting are wedded to the notion that cue-overload interference procedures (often involving the A-B, A-C list-learning paradigm) capture the most important elements of forgetting in everyday life. However, findings from a century of work in psychology, psychopharmacology, and neuroscience converge on the notion that such procedures may pertain mainly to forgetting in the laboratory and that everyday forgetting is attributable to an altogether different form of interference. According to this idea, recently formed memories that have not yet had a chance to consolidate are vulnerable to the interfering force of mental activity and memory formation (even if the interfering activity is not similar to the previously learned material). This account helps to explain why sleep, alcohol, and benzodiazepines all improve memory for a recently learned list, and it is consistent with recent work on the variables that affect the induction and maintenance of long-term potentiation in the hippocampus. PMID- 14744217 TI - Object perception as Bayesian inference. AB - We perceive the shapes and material properties of objects quickly and reliably despite the complexity and objective ambiguities of natural images. Typical images are highly complex because they consist of many objects embedded in background clutter. Moreover, the image features of an object are extremely variable and ambiguous owing to the effects of projection, occlusion, background clutter, and illumination. The very success of everyday vision implies neural mechanisms, yet to be understood, that discount irrelevant information and organize ambiguous or noisy local image features into objects and surfaces. Recent work in Bayesian theories of visual perception has shown how complexity may be managed and ambiguity resolved through the task-dependent, probabilistic integration of prior object knowledge with image features. PMID- 14744218 TI - Development in midlife. AB - The midlife period in the lifespan is characterized by a complex interplay of multiple roles. The goal of this chapter is to summarize research findings on the central themes and salient issues of midlife such as balancing work and family responsibilities in the midst of the physical and psychological changes associated with aging. The field of midlife development is emerging in the context of large demographic shifts in the population. A section on the phenomenology of midlife development presents images and expectations including the seemingly disparate views of midlife as a time of peak functioning and a period of crisis. Conceptual frameworks useful for studying the multiple patterns of change in midlife are presented. Findings demonstrating patterns of gains and losses are reviewed for multiple domains: cognitive functioning, personality and the self, emotions, social relationships, work, and physical health. The need for future research to illuminate and integrate the diverse aspects of midlife is highlighted. PMID- 14744219 TI - The intergenerational transfer of psychosocial risk: mediators of vulnerability and resilience. AB - The recurrence of social, behavioral, and health problems in successive generations of families is a prevalent theme in both the scientific and popular literatures. This review discusses recent conceptual models and findings from longitudinal studies concerning the intergenerational transfer of psychosocial risk, including intergenerational continuity, and the processes whereby a generation of parents may place their offspring at elevated risk for social, behavioral, and health problems. Key findings include the mediational effects of parenting and environmental factors in the transfer of risk. In both girls and boys, childhood aggression and antisocial behavior appear to predict long-term trajectories that place offspring at risk. Sequelae of childhood aggression that may threaten the well-being of offspring include school failure, adolescent risk taking behavior, early and single parenthood, and family poverty. These childhood and adolescent behavioral styles also predict harsh, aggressive, neglectful, and unstimulating parenting behavior toward offspring. Buffering factors within at risk families include maternal educational attainment and constructive parenting practices (e.g., emotional warmth, consistent disciplinary practices, and cognitive scaffolding). These findings highlight the potential application and relevance of intergenerational studies for social, educational, and health policy. PMID- 14744220 TI - Development in the family. AB - In this chapter we review theoretical conceptual and empirical advances in family research and the implications for children's development. Three interdependent family subsystems are considered: the parent-child subsystem, the marital subsystem, and the sibling subsystem. Recent work on the family as a unit of analysis is reviewed as well. Several determinants of family socialization strategies are noted, including child and parental characteristics and resources and social capital available to families. Ethnicity is considered by reviewing recent advances in our understanding of African American, Asian American, and Hispanic families. Several new directions in family research are highlighted. These include research on the linkage between families and work, gay and lesbian parenting, the new reproductive technologies, and the effect of parental incarcerations on families. Future trends in family research are noted, including the role of genetics, the biological correlates of family processes, the role of intervention and prevention designs in family research, and the importance of process work on ethnicity-oriented family research. PMID- 14744221 TI - Schizophrenia: etiology and course. AB - Decades of research on schizophrenia have not produced major breakthroughs, but gradual progress has been made in identifying risk factors and clarifying the nature of the etiologic process. This article provides an overview of trends in research findings as well as current assumptions about the interplay between environmental and genetic factors in the etiology of schizophrenia. Based on the cumulative findings, it appears that both genetic and prenatal factors can give rise to constitutional vulnerability. Subsequent neuromaturational processes, especially those that occur during adolescence, and exposure to stressful events can trigger the behavioral expression of this vulnerability. PMID- 14744222 TI - Clinical implications of reinforcement as a determinant of substance use disorders. AB - Extensive scientific evidence indicates that reinforcement plays an important role in the genesis, maintenance, and recovery from substance use disorders. In this chapter, we review recent clinical research from laboratory, clinic, and naturalistic settings examining the role of reinforcement in substance use disorders. Well-controlled human laboratory studies are reviewed characterizing orderly interactions between the reinforcing effects of drugs and environmental context that have important implications for understanding risk factors for substance use disorders and for the development of efficacious interventions. Recent treatment-outcome studies on voucher-based contingency management and community reinforcement therapy are reviewed demonstrating how reinforcement and related principles can be used to improve outcomes across a wide range of different substance use disorders and populations. Overall, the chapter characterizes a vigorous area of clinical research that has much to contribute to a scientific analysis of substance use disorders. PMID- 14744223 TI - Motivational influences on cigarette smoking. AB - Cigarette smoking is a leading cause of mortality and morbidity and a particularly common and intractable addictive disorder. Research shows that nicotine is a sine qua non of tobacco addiction and that it produces the hallmark effects of addictive drugs: sensitization, tolerance, physical dependence, and euphoria/elation. Research on the development of smoking reveals that although smoking prevalence has declined from a peak in the mid-1990s, close to 30% of twelfth graders still smoke. Smoking in adolescents is related to development of physical dependence, ethnicity, impulsivity, affective disorder, and peer influences. However, which of these exerts the greatest causal effects is unknown, and their influence no doubt varies across individuals and across development. Once dependence on tobacco smoking is established, evidence suggests that tobacco motivation is strongly influenced by a reduction in withdrawal symptoms, an expectation of stress reduction, and conditioned reinforcement. Nicotine motivation may also be influenced by modulation in stimulus incentive value. PMID- 14744225 TI - Gender in psychology. AB - Gender is increasingly understood as defining a system of power relations embedded in other power relations. Psychological research on gender-which has most often focused on analysis of sex differences, within-sex variability, and gender roles-has begun to incorporate this new understanding. By drawing on three resources, psychologists can make more rapid progress in understanding gender's significance for psychological processes: social science theories that link the individual and social levels of analysis; constructs (such as identity) that bridge the social and individual levels; and conceptual tools generated in feminist theory, perhaps especially intersectionality. We review these resources, cite active research programs that have employed them, and conclude by offering some practical suggestions about how to incorporate these resources into our research. PMID- 14744224 TI - Self-knowledge: its limits, value, and potential for improvement. AB - Because of personal motives and the architecture of the mind, it may be difficult for people to know themselves. People often attempt to block out unwanted thoughts and feelings through conscious suppression and perhaps through unconscious repression, though whether such attempts are successful is controversial. A more common source of self-knowledge failure is the inaccessibility of much of the mind to consciousness, including mental processes involved in perception, motor learning, personality, attitudes, and self-esteem. Introspection cannot provide a direct pipeline to these mental processes, though some types of introspection may help people construct beneficial personal narratives. Other ways of increasing self-knowledge include looking at ourselves through the eyes of others and observing our own behavior. These approaches can potentially promote self-knowledge, although major obstacles exist. It is not always advantageous to hold self-perceptions that correspond perfectly with reality, but increasing awareness of nonconscious motives and personality is generally beneficial. PMID- 14744226 TI - Mediated politics and citizenship in the twenty-first century. AB - Since the birth of the nation, concepts about the political duties of citizens have changed drastically to keep pace with growth and development. The information needs have changed as well, as have the institutions that supply this information. In this essay I analyze the interrelation between citizenship in the twenty-first century and the information supply that nourishes it. I focus on studies that explore how political news is shaped to attract public attention and how citizens select it and make sense of it. Evidence from content analyses, focus group data, and intensive interviews supports the conclusion that the news supply is adequate for citizens' civic needs and that they use it judiciously. To accept that conclusion requires abandoning outdated paradigms of citizenship that ignore information-processing capabilities of human beings, the basic motivations that drive the search for political information, and the impact of the ever increasing complexity of politics. PMID- 14744227 TI - The internet and social life. AB - The Internet is the latest in a series of technological breakthroughs in interpersonal communication, following the telegraph, telephone, radio, and television. It combines innovative features of its predecessors, such as bridging great distances and reaching a mass audience. However, the Internet has novel features as well, most critically the relative anonymity afforded to users and the provision of group venues in which to meet others with similar interests and values. We place the Internet in its historical context, and then examine the effects of Internet use on the user's psychological well-being, the formation and maintenance of personal relationships, group memberships and social identity, the workplace, and community involvement. The evidence suggests that while these effects are largely dependent on the particular goals that users bring to the interaction-such as self-expression, affiliation, or competition-they also interact in important ways with the unique qualities of the Internet communication situation. PMID- 14744228 TI - Social influence: compliance and conformity. AB - This review covers recent developments in the social influence literature, focusing primarily on compliance and conformity research published between 1997 and 2002. The principles and processes underlying a target's susceptibility to outside influences are considered in light of three goals fundamental to rewarding human functioning. Specifically, targets are motivated to form accurate perceptions of reality and react accordingly, to develop and preserve meaningful social relationships, and to maintain a favorable self-concept. Consistent with the current movement in compliance and conformity research, this review emphasizes the ways in which these goals interact with external forces to engender social influence processes that are subtle, indirect, and outside of awareness. PMID- 14744229 TI - Group performance and decision making. AB - Theory and research on small group performance and decision making is reviewed. Recent trends in group performance research have found that process gains as well as losses are possible, and both are frequently explained by situational and procedural contexts that differentially affect motivation and resource coordination. Research has continued on classic topics (e.g., brainstorming, group goal setting, stress, and group performance) and relatively new areas (e.g., collective induction). Group decision making research has focused on preference combination for continuous response distributions and group information processing. New approaches (e.g., group-level signal detection) and traditional topics (e.g., groupthink) are discussed. New directions, such as nonlinear dynamic systems, evolutionary adaptation, and technological advances, should keep small group research vigorous well into the future. PMID- 14744230 TI - Creativity. AB - Creativity has clear benefits for individuals and society as a whole. Not surprisingly, a great deal of research has focused on creativity, especially in the past 20 years. This chapter reviews the creativity research, first looking to the relevant traits, capacities, influences, and products, and then within disciplinary perspectives on creativity (e.g., biological, cognitive, developmental, organizational). Great headway is being made in creativity research, but more dialogue between perspectives is suggested. New and important areas of research are highlighted, and the various costs and benefits of creativity are discussed. PMID- 14744231 TI - Psychology and culture. AB - Psychological processes influence culture. Culture influences psychological processes. Individual thoughts and actions influence cultural norms and practices as they evolve over time, and these cultural norms and practices influence the thoughts and actions of individuals. Large bodies of literature support these conclusions within the context of research on evolutionary processes, epistemic needs, interpersonal communication, attention, perception, attributional thinking, self-regulation, human agency, self-worth, and contextual activation of cultural paradigms. Cross-cultural research has greatly enriched psychology, and key issues for continued growth and maturation of the field of cultural psychology are articulated. PMID- 14744232 TI - Teaching of subject matter. AB - Psychology of subject matter refers to the scientific study of learning and instruction within school subjects. The growing research literature on teaching and learning of school subjects represents one of educational psychology's most productive accomplishments of the past two decades. The purpose of this chapter is to examine representative advances in the psychology of subject matter, including how people learn to read words, comprehend printed passages, write compositions, solve arithmetic word problems, and understand how scientific systems work. The introduction provides a historical overview of how to promote transfer and is followed by reviews of representative research in learning and teaching of reading fluency, reading comprehension, writing, mathematics, and science. PMID- 14744233 TI - Coping: pitfalls and promise. AB - Coping, defined as the thoughts and behaviors used to manage the internal and external demands of situations that are appraised as stressful, has been a focus of research in the social sciences for more than three decades. The dramatic proliferation of coping research has spawned healthy debate and criticism and offered insight into the question of why some individuals fare better than others do when encountering stress in their lives. We briefly review the history of contemporary coping research with adults. We discuss three primary challenges for coping researchers (measurement, nomenclature, and effectiveness), and highlight recent developments in coping theory and research that hold promise for the field, including previously unaddressed aspects of coping, new measurement approaches, and focus on positive affective outcomes. PMID- 14744234 TI - Survey research and societal change. AB - Surveys reflect societal change in a way that few other research tools do. Over the past two decades, three developments have transformed surveys. First, survey organizations have adopted new methods for selecting telephone samples; these new methods were made possible by the creation of large databases that include all listed telephone numbers in the United States. A second development has been the widespread decline in response rates for all types of surveys. In the face of this problem, survey researchers have developed new theories of nonresponse that build on the persuasion literature in social psychology. Finally, surveys have adopted many new methods of data collection; the new modes reflect technological developments in computing and the emergence of the Internet. Research has spawned several theories that examine how characteristics of the data collection method shape the answers obtained. Rapid change in survey methods is likely to continue in the coming years. PMID- 14744235 TI - Human research and data collection via the internet. AB - Advantages and disadvantages of Web and lab research are reviewed. Via the World Wide Web, one can efficiently recruit large, heterogeneous samples quickly, recruit specialized samples (people with rare characteristics), and standardize procedures, making studies easy to replicate. Alternative programming techniques (procedures for data collection) are compared, including client-side as opposed to server-side programming. Web studies have methodological problems; for example, higher rates of drop out and of repeated participation. Web studies must be thoroughly analyzed and tested before launching on-line. Many studies compared data obtained in Web versus lab. These two methods usually reach the same conclusions; however, there are significant differences between college students tested in the lab and people recruited and tested via the Internet. Reasons that Web researchers are enthusiastic about the potential of the new methods are discussed. PMID- 14744236 TI - Predicting human drug glucuronidation parameters: application of in vitro and in silico modeling approaches. AB - Cytochrome P450 (CYP) and UDP-glucuronosyltransferase (UGT), which both exist as enzyme "superfamilies," are together responsible for the metabolism of most hepatically cleared drugs. There is currently intense interest in the development of techniques that permit identification of the CYP and UGT isoform(s) involved in the metabolism of a newly discovered drug, and hence prediction of factors likely to alter elimination in vivo. In addition, the quantitative scaling of kinetic parameters for a metabolic pathway assumes importance for identifying newly discovered drugs with undesirable in vivo pharmacokinetic properties. Although qualitative and quantitative in vitro-in vivo correlation based on data generated using human liver tissue or recombinant enzymes have been applied successfully to many drugs eliminated by CYP, these strategies have proved less definitive for glucuronidated compounds. Computational (in silico) modeling techniques that potentially provide a facile and economic alternative to the in vitro methods are now emerging. This review assesses the utility of in vitro and in silico approaches for the qualitative and quantitative prediction of drug glucuronidation parameters and the challenges facing the development of generalizable models. PMID- 14744237 TI - Oxidative stress, toxicology, and pharmacology of CYP2E1. AB - This review describes some of the biochemical and toxicological properties of CYP2E1, especially as it relates to alcohol metabolism and toxicity and the establishment of human hepatoma HepG2 cell lines that overexpress human CYP2E1. Ethanol, polyunsaturated fatty acids, and iron were found to be cytotoxic in HepG2 cells that overexpress CYP2E1. GSH appears to be essential in protecting HepG2 cells against the CYP2E1-dependent cytotoxicity, and GSH levels were elevated owing to a twofold increase in activity and expression of glutamate cysteine ligase. We suggest that this up-regulation of GSH synthesis was an adaptive response to attenuate CYP2E1-dependent oxidative stress and toxicity. Induction of a state of oxidative stress appears to play a central role in the CYP2E1-dependent cytotoxicity. Mitochondrial membrane potential decreased in the CYP2E1-expressing HepG2 cells, and this decrease shared similar characteristics with the developing toxicity. Alcohol-dependent liver injury is likely to be a multifactorial process involving several mechanisms. We believe that the linkage between CYP2E1-dependent oxidative stress, mitochondrial injury, and GSH homeostasis contribute to the toxic actions of ethanol on the liver. PMID- 14744238 TI - The identification of ligands at orphan G-protein coupled receptors. AB - The completion of the human genome sequencing project has identified approximately 720 genes that belong to the G-protein coupled receptor (GPCR) superfamily. Approximately half of these genes are thought to encode sensory receptors. Of the remaining 360 receptors, the natural ligand has been identified for approximately 210 receptors, leaving 150 so-called orphan GPCRs with no known ligand or function. The identification of ligands active at orphan GPCRs has been achieved through the development of a number of experimental approaches, including the screening of putative small molecule and peptide ligands, reverse pharmacology, and the use of bioinformatics to predict candidate ligands. In this review, we discuss the methodologies developed for the identification of ligands at orphan GPCRs and include examples of their successful application. PMID- 14744239 TI - Biochemical mechanism of nitroglycerin action and tolerance: is this old mystery solved? AB - Organic nitrates such as nitroglycerin (NTG) have been used as potent vasodilators in medicine for more than a century, but their biochemical mechanisms of action, particularly in relation to tolerance development, are still incompletely defined. Numerous candidate enzymes for NTG metabolism, as well as a multiplicity of tolerance mechanisms, have been proposed in the literature, but a consolidating hypothesis that links these phenomena together has not appeared. Here, we outline a "thionitrate oxidation hypothesis," which attempts to link nitrate bioactivation and tolerance development in an overall mechanism. We also attempt to compare and contrast the proposed mechanism against existing theories of nitrate action and tolerance. Interactions between organic nitrates, which have been thought of as endothelium-independent agents, and the vascular endothelium and endothelial nitric oxide synthase (eNOS) are also discussed. PMID- 14744241 TI - The integration of pharmacokinetics and pharmacodynamics: understanding dose response. AB - Pharmacokinetic (PK) and pharmacodynamic (PD) studies have proven to be powerful and instructive tools, particularly in elucidating important aspects of human pharmacology. Nevertheless, they remain imperfect tools in that they only allow researchers to indirectly extrapolate, through computational modeling, the dynamic processes of drug action. Furthermore, neither tool alone provides a complete nor necessarily relevant picture of drug action. This review explores the utility and applications of PK and PD in the study of drugs, provides examples of lessons learned from their application to studies of human pharmacology, points out some of their limitations, and advances the thesis that these tools ideally should be employed together in an integrated approach. As we continue to apply these tools across the continuum of age and disease, they provide a powerful means to enhance our understanding of drug action, drug interactions, and intrinsic host factors that influence pharmacologic response. PMID- 14744242 TI - Transporters and renal drug elimination. AB - Carrier-mediated processes, often referred to as transporters, play key roles in the reabsorption and secretion of many endogenous and xenobiotic compounds by the kidney. The renal proximal tubule is the primary site of active transport for a wide variety of substrates, including organic anions/cations, peptides, and nucleosides. During the past decade, significant advances in molecular identification and characterization of transporter proteins have been made. Although it is generally noted that these transporters significantly contribute to renal drug handling and variability in drug disposition, the extent of our knowledge regarding the specific roles of such transporters in drug disposition and drug-drug interactions remains, for the most part, limited. In this review, we summarize recent progress in terms of molecular and functional characterization of renal transporters and their clinical relevance to drug therapy. PMID- 14744240 TI - Developmental neuropathology of environmental agents. AB - The developing central nervous system (CNS) is more vulnerable to injury than the adult one. Although a great deal of research has been devoted to subtle effects of developmental exposure, such as neurobehavioral changes, this review instead focuses on a number of chemicals that have been shown, in several experimental models as well as humans, to cause morphological changes in the developing nervous system. Chemicals that are discussed include methylmercury (MeHg), lead (Pb), antiepileptic drugs, and ethanol. Additionally, the issue of silent neurotoxicity, i.e., persistent morphological and/or biochemical injury that remains clinically unapparent until later in life, is discussed. PMID- 14744243 TI - Identification of the major steps in botulinum toxin action. AB - Botulinum toxin is a uniquely potent substance synthesized by the organisms Clostridium botulinum, Clostridium baratii, and Clostridium butyricum. This toxin, which acts preferentially on peripheral cholinergic nerve endings to block acetylcholine release, is both an agent that causes disease (i.e., botulism) as well as an agent that can be used to treat disease (e.g., dystonia). The ability of botulinum toxin to produce its effects is largely dependent on its ability to penetrate cellular and intracellular membranes. Thus, toxin that is ingested or inhaled can bind to epithelial cells and be transported to the general circulation. Toxin that reaches peripheral nerve endings binds to the cell surface then penetrates the plasma membrane by receptor-mediated endocytosis and the endosome membrane by pH-induced translocation. Internalized toxin acts in the cytosol as a metalloendoprotease to cleave polypeptides that are essential for exocytosis. This review seeks to identify and characterize all major steps in toxin action, from initial absorption to eventual paralysis of cholinergic transmission. PMID- 14744244 TI - ErbB receptors: directing key signaling networks throughout life. AB - The epidermal growth factor (EGF)-related peptides bind the ErbB receptors, inducing the formation of different homo- and heterodimers. Receptor dimerization promotes activation of the intrinsic kinase, leading to phosphorylation of specific tyrosines located in the ErbB's cytoplasmic region. These phosphorylated residues serve as docking sites for a variety of signaling molecules whose recruitment stimulates intracellular signaling cascades, which ultimately control diverse genetic programs. Particular ligand-receptor complexes have essential roles in embryonic development as well as in the adult. Finally, ErbB receptors are being pursued as therapeutic targets because aberrant ErbB activity has been observed in many human cancers. In this review, we discuss these data in more detail, illustrating the importance of tightly regulated ErbB signaling throughout life. PMID- 14744245 TI - Novel angiogenic signaling pathways and vascular targets. AB - Intense investigation into the molecular basis of angiogenesis is rapidly revealing novel signaling pathways involved in the generation of new vasculature. These range from elucidation of the mechanism by which hypoxia initiates expression of a proangiogenic gene repertoire via the hypoxia-inducible transcription factors (HIFs) to molecular pathways involved in extra- and intracellular signaling during new vessel formation. Extracellular pathways include those of the Notch/delta, ephrin/Eph receptor and roundabout/slit families, and intracellular pathway members of the hedgehog and sprouty families. The involvement of these pathways in angiogenesis is discussed, together with some comments on recently identified targets in the vasculature that present new therapeutic opportunities. PMID- 14744246 TI - The role of oxidative stress in carcinogenesis. AB - Chemical carcinogenesis follows a multistep process involving both mutation and increased cell proliferation. Oxidative stress can occur through overproduction of reactive oxygen and nitrogen species through either endogenous or exogenous insults. Important to carcinogenesis, the unregulated or prolonged production of cellular oxidants has been linked to mutation (induced by oxidant-induced DNA damage), as well as modification of gene expression. In particular, signal transduction pathways, including AP-1 and NFkappaB, are known to be activated by reactive oxygen species, and they lead to the transcription of genes involved in cell growth regulatory pathways. This review examines the evidence of cellular oxidants' involvement in the carcinogenesis process, and focuses on the mechanisms for production, cellular damage produced, and the role of signaling cascades by reactive oxygen species. PMID- 14744247 TI - DARPP-32: an integrator of neurotransmission. AB - Dopamine- and cAMP-regulated phosphoprotein, Mr 32 kDa (DARPP-32), was identified initially as a major target for dopamine and protein kinase A (PKA) in striatum. However, recent advances now indicate that regulation of the state of DARPP-32 phosphorylation provides a mechanism for integrating information arriving at dopaminoceptive neurons, in multiple brain regions, via a variety of neurotransmitters, neuromodulators, neuropeptides, and steroid hormones. Activation of PKA or PKG stimulates DARPP-32 phosphorylation at Thr34 and thereby converts DARPP-32 into a potent inhibitor of protein phosphatase-1 (PP-1). DARPP 32 is also phosphorylated at Thr75 by Cdk5 and this converts DARPP-32 into an inhibitor of PKA. Thus, DARPP-32 has the unique property of being a dual-function protein, acting either as an inhibitor of PP-1 or of PKA. The state of phosphorylation of DARPP-32 at Thr34 depends on the phosphorylation state of two serine residues, Ser102 and Ser137, which are phosphorylated by CK2 and CK1, respectively. By virtue of its ability to modulate the activity of PP-1 and PKA, DARPP-32 is critically involved in regulating electrophysiological, transcriptional, and behavioral responses to physiological and pharmacological stimuli, including antidepressants, neuroleptics, and drugs of abuse. PMID- 14744248 TI - Beta-adrenergic receptors and regulation of energy expenditure: a family affair. AB - The family of adrenergic receptors (ARs) expressed in adipocytes includes three sibling betaARs and two alphaAR cousins. Together they profoundly influence the mobilization of stored fatty acids, secretion of fat-cell derived hormones, and the specialized process of nonshivering thermogenesis in brown adipose tissue. The two types of fat cells that compose adipose tissue, brown and white, are structurally and functionally distinct. Studies on the mechanisms by which individual betaAR regulates these cell-specific functions have recently uncovered new signal transduction cascades involved in processes traditionally ascribed to adenylyl cyclase/cAMP/protein kinase A system. They illustrate how betaAR signaling can orchestrate a coordinated set of intracellular responses for fine control of metabolic balance. PMID- 14744249 TI - Protein sulfenic acids in redox signaling. AB - Reactive (low pKa) cysteine residues in proteins are critical components in redox signaling. A particularly reactive and versatile reversibly oxidized form of cysteine, the sulfenic acid (Cys-SOH), has important roles as a catalytic center in enzymes and as a sensor of oxidative and nitrosative stress in enzymes and transcriptional regulators. Depending on environment, sometimes the sulfenic acid provides a metastable oxidized form, and other times it is a fleeting intermediate giving rise to more stable disulfide, sulfinic acid, or sulfenyl amide forms. PMID- 14744250 TI - The role of calpain in oncotic cell death. AB - Numerous lines of evidence demonstrate that calpains, a family of 14 Ca(2+) activated neutral cysteine proteases, are involved in oncotic cell death in a variety of models. At this time, the biochemistry of most calpains and the specific roles of different calpains in physiology and pathology remain to be determined. A number of calpain substrates have been identified in cellular systems, including cytoskeletal proteins, and recent studies suggest that calpains mediate the increase in plasma membrane permeability to ions and the progressive breakdown of the plasma membrane observed in oncosis through the proteolysis of cystokeletal and plasma membrane proteins. Further, a number of reports provide evidence that the mitochondrial dysfunction observed in oncosis may be mediated by a mitochondrial calpain of unknown identity. Finally, a number of diverse calpain inhibitors have been developed that show cytoprotective properties in cellular systems and in vivo following diverse insults. It is suggested that future research be directed toward elucidation of the role(s) of specific calpain isozymes in physiological and pathological conditions; identifying and linking specific calpain substrates with altered cellular functions; and developing cell-permeable, potent, isozyme-selective calpain inhibitors. PMID- 14744251 TI - Voltage-gated sodium channels and hyperalgesia. AB - Physiological and pharmacological evidence both have demonstrated a critical role for voltage-gated sodium channels (VGSCs) in many types of chronic pain syndromes because these channels play a fundamental role in the excitability of neurons in the central and peripheral nervous systems. Alterations in function of these channels appear to be intimately linked to hyperexcitability of neurons. Many types of pain appear to reflect neuronal hyperexcitability, and importantly, use dependent sodium channel blockers are effective in the treatment of many types of chronic pain. This review focuses on the role of VGSCs in the hyperexcitability of sensory primary afferent neurons and their contribution to the inflammatory or neuropathic pain states. The discrete localization of the tetrodotoxin (TTX) resistant channels, in particular NaV1.8, in the peripheral nerves may provide a novel opportunity for the development of a drug targeted at these channels to achieve efficacious pain relief with an acceptable safety profile. PMID- 14744252 TI - Neurogenesis in the adult brain: new strategies for central nervous system diseases. AB - New cells are continuously generated from immature proliferating cells throughout adulthood in many organs, thereby contributing to the integrity of the tissue under physiological conditions and to repair following injury. In contrast, repair mechanisms in the adult central nervous system (CNS) have long been thought to be very limited. However, recent findings have clearly demonstrated that in restricted areas of the mammalian brain, new functional neurons are constantly generated from neural stem cells throughout life. Moreover, stem cells with the potential to give rise to new neurons reside in many different regions of the adult CNS. These findings raise the possibility that endogenous neural stem cells can be mobilized to replace dying neurons in neurodegenerative diseases. Indeed, recent reports have provided evidence that, in some injury models, limited neuronal replacement occurs in the CNS. Here, we summarize our current understanding of the mechanisms controlling adult neurogenesis and discuss their implications for the development of new strategies for the treatment of neurodegenerative diseases. PMID- 14744253 TI - Muscarinic acetylcholine receptor knockout mice: novel phenotypes and clinical implications. AB - Muscarinic acetylcholine receptors (mAChRs; M1-M5) play key roles in regulating the activity of many important functions of the central and peripheral nervous system. Because of the lack of ligands endowed with a high degree of receptor subtype selectivity and the fact that most tissues or cell types express two or more mAChR subtypes, identification of the physiological and pathophysiological roles of the individual mAChR subtypes has proven a difficult task. To circumvent these difficulties, several laboratories recently employed gene-targeting techniques to generate mutant mouse strains deficient in each of the five mAChR subtypes. Phenotyping studies showed that each mutant mouse line displayed characteristic physiological, pharmacological, behavioral, biochemical, or neurochemical deficits. The novel insights gained from these studies should prove instrumental for the development of novel classes of muscarinic drugs. PMID- 14744254 TI - Mixed-lineage kinases: a target for the prevention of neurodegeneration. AB - The activation of the c-Jun N-terminal kinase (JNK) pathway is critical for naturally occurring neuronal cell death during development and may be important for the pathological neuronal cell death of neurodegenerative diseases. The small molecule inhibitor of the mixed-lineage kinase (MLK) family of kinases, CEP-1347, inhibits the activation of the JNK pathway and, consequently, the cell death in many cell culture and animal models of neuronal death. CEP-1347 has the ability not only to inhibit cell death but also to maintain the trophic status of neurons in culture. The possible importance of the JNK pathway in neurodegenerative diseases such as Alzheimer's and Parkinson's diseases provides a rationale for the use of CEP-1347 for the treatment of these diseases. CEP-1347 has the potential of not only retarding disease progression but also reversing the severity of symptoms by improving the function of surviving neurons. PMID- 14744255 TI - Analysis of GABAA receptor function and dissection of the pharmacology of benzodiazepines and general anesthetics through mouse genetics. AB - GABAA receptors are molecular substrates for the regulation of vigilance, anxiety, muscle tension, epileptogenic activity, and memory functions, and the enhancement of GABAA receptor-mediated fast synaptic inhibition is the basis for the pharmacotherapy of various neurological and psychiatric disorders. Two kinds of GABAA receptor-targeted mutant mice have been generated: (a) knockout mice that lack individual GABAA receptor subunits (alpha1, alpha5, alpha6, beta2, beta3, gamma2, delta, and rho1) and (b) knockin mice that carry point mutations affecting the action of modulatory drugs [alpha1(H101R), alpha2(H101R), alpha3(H126R), alpha5(H105R), and beta3(N265M)]. Whereas the knockout mice have provided information primarily with respect to the regulation of subunit gene transcription, receptor assembly, and some physiological functions of individual receptor subtypes, the point-mutated knockin mice in which specific GABAA receptor subtypes are insensitive to diazepam or some general anesthetics have revealed the specific contribution of individual receptor subtypes to the pharmacological spectrum of diazepam and general anesthetics. PMID- 14744256 TI - Sex differences in pharmacokinetics and pharmacodynamics. AB - The importance of reviewing and studying sex-based differences in pharmacologic parameters is demonstrated by the increasing data on gender variation in drug efficacy and toxicity profiles. Sex-based differences in the four major factors that contribute to interindividual pharmacokinetic variability--bioavailability, distribution, metabolism, and elimination--are theorized to stem from variations between men and women in factors such as body weight, plasma volume, gastric emptying time, plasma protein levels, cytochrome P450 activity, drug transporter function, and excretion activity. Sex-determined variations in pharmacodynamics have traditionally been more difficult to study, but a number of recent studies have explored these differences. This review examines the biologic basis of differences in pharmacokinetics and pharmacodynamics between the sexes and summarizes studies that have addressed these differences. As an example, sex based variation in the efficacy and toxicity of antiretroviral therapy in human immunodeficiency virus (HIV)-infected patients is explored more thoroughly to illustrate some of the factors underlying sex-based differences in drug therapy. PMID- 14744257 TI - CRF and CRF receptors: role in stress responsivity and other behaviors. AB - Since corticotropin-releasing factor (CRF) was first characterized, a growing family of ligands and receptors has evolved. The mammalian family members include CRF, urocortinI (UcnI), UcnII, and UcnIII, along with two receptors, CRFR1 and CRFR2, and a CRF binding protein. These family members differ in their tissue distribution and pharmacology. Studies have provided evidence supporting an important role of this family in regulation of the endocrine and behavioral responses to stress. Although CRF appears to play a stimulatory role in stress responsivity through activation of CRFR1, specific actions of UcnII and UcnIII on CRFR2 may be important for dampening stress sensitivity. As the only ligand with high affinity for both receptors, UcnI's role may be promiscuous. Regulation of the relative contribution of the two CRF receptors to brain CRF pathways may be essential in coordinating physiological responses to stress. The development of disorders related to heightened stress sensitivity and dysregulation of stress coping mechanisms appears to involve regulatory mechanisms of CRF family members. PMID- 14744258 TI - Membrane trafficking of G protein-coupled receptors. AB - G protein-coupled receptors (GPCRs) modulate diverse physiological and behavioral signaling pathways by virtue of changes in receptor activation and inactivation states. Functional changes in receptor properties include dynamic interactions with regulatory molecules and trafficking to various cellular compartments at various stages of the life cycle of a GPCR. This review focuses on trafficking of GPCRs to the cell surface, stabilization there, and agonist-regulated turnover. GPCR interactions with a variety of newly revealed partners also are reviewed with the intention of provoking further analysis of the relevance of these interactions in GPCR trafficking, signaling, or both. The disease consequences of mislocalization of GPCRs also are described. PMID- 14744259 TI - 14-3-3-affinity purification of over 200 human phosphoproteins reveals new links to regulation of cellular metabolism, proliferation and trafficking. AB - 14-3-3-interacting proteins were isolated from extracts of proliferating HeLa cells using 14-3-3 affinity chromatography, eluting with a phosphopeptide that competes with targets for 14-3-3 binding. The isolated proteins did not bind to 14-3-3 proteins (14-3-3s) after dephosphorylation with protein phosphatase 2A (PP2A), indicating that binding to 14-3-3s requires their phosphorylation. The binding proteins identified by tryptic mass fingerprinting and Western blotting include many enzymes involved in generating precursors such as purines (AMP, GMP and ATP), FAD, NADPH, cysteine and S-adenosylmethionine, which are needed for cell growth, regulators of cell proliferation, including enzymes of DNA replication, proteins of anti-oxidative metabolism, regulators of actin dynamics and cellular trafficking, and proteins whose deregulation has been implicated in cancers, diabetes, Parkinsonism and other neurological diseases. Several proteins bound to 14-3-3-Sepharose in extracts of proliferating cells, but not in non proliferating, serum-starved cells, including a novel microtubule-interacting protein ELP95 (EMAP-like protein of 95 kDa) and a small HVA22/Yop1p-related protein. In contrast, the interactions of 14-3-3s with the N-methyl-D-aspartate receptor 2A subunit and NuMA (nuclear mitotic apparatus protein) were not regulated by serum. Overall, our findings suggest that 14-3-3s may be central to integrating the regulation of biosynthetic metabolism, cell proliferation, survival, and other processes in human cells. PMID- 14744260 TI - Factors determining the oxygen consumption rate (VO2) on-kinetics in skeletal muscles. AB - Using a computer model of oxidative phosphorylation developed previously [Korzeniewski and Mazat (1996) Biochem. J. 319, 143-148; Korzeniewski and Zoladz (2001) Biophys. Chem. 92, 17-34], we analyse the effect of several factors on the oxygen-uptake kinetics, especially on the oxygen consumption rate (VO2) and half transition time t(1/2), at the onset of exercise in skeletal muscles. Computer simulations demonstrate that an increase in the total creatine pool [PCr+/-Cr] (where Cr stands for creatine and PCr for phosphocreatine) and in glycolytic ATP supply lengthen the half-transition time, whereas increase in mitochondrial content, in parallel activation of ATP supply and ATP usage, in oxygen concentration, in proton leak, in resting energy demand, in resting cytosolic pH and in initial alkalization decrease this parameter. Theoretical studies show that a decrease in the activity of creatine kinase (CK) [displacement of this enzyme from equilibrium during on-transient (rest-to-work transition)] accelerates the first stage of the VO2 on-transient, but slows down the second stage of this transient. It is also demonstrated that a prior exercise terminated a few minutes before the principal exercise shortens the transition time. Finally, it is shown that at a given ATP demand, and under conditions where CK works near the thermodynamic equilibrium, the half-transition time of VO2 kinetics is determined by the amount of PCr that has to be transformed into Cr during rest-to-work transition; therefore any factor that diminishes the difference in [PCr] between rest and work at a given energy demand will accelerate the VO2 on-kinetics. Our conclusions agree with the general idea formulated originally by Easterby [(1981) Biochem. J. 199, 155-161] that changes in metabolite concentrations determine the transition times between different steady states in metabolic systems. PMID- 14744261 TI - Differential modulation of androgen receptor transcriptional activity by the nuclear receptor co-repressor (N-CoR). AB - Antiandrogens are widely used agents in the treatment of prostate cancer, as inhibitors of AR (androgen receptor) action. Although the precise mechanism of antiandrogen action is not yet elucidated, recent studies indicate the involvement of nuclear receptor co-repressors. In the present study, the regulation of AR transcriptional activity by N-CoR (nuclear receptor co repressor), in the presence of different ligands, has been investigated. Increasing levels of N-CoR differentially affected the transcriptional activity of AR occupied with either agonistic or antagonistic ligands. Small amounts of co transfected N-CoR repressed CPA (cyproterone acetate)- and mifepristone (RU486) mediated AR activity, but did not affect agonist (R1881)-induced AR activity. Larger amounts of co-transfected N-CoR repressed AR activity for all ligands, and converted the partial agonists CPA and RU486 into strong AR antagonists. In the presence of the agonist R1881, co-expression of the p160 co-activator TIF2 (transcriptional intermediary factor 2) relieved N-CoR repression up to control levels. However, in the presence of RU486 and CPA, TIF2 did not functionally compete with N-CoR, suggesting that antagonist-bound AR has a preference for N CoR. The AR mutation T877A (Thr877-->Ala), which is frequently found in prostate cancer and affects the ligand-induced conformational change of the AR, considerably reduced the repressive action of N-CoR. The agonistic activities of CPA- and hydroxyflutamide-occupied T877A-AR were hardly affected by N-CoR, whereas TIF2 strongly enhanced their activities. These results indicate that lack of N-CoR action allows these antiandrogens to act as strong agonists on the mutant AR. PMID- 14744263 TI - Demand valves for oxygen therapy: your mileage may vary. PMID- 14744262 TI - Enzymic characterization with progress curve analysis of a collagen peptidase from an enthomopathogenic bacterium, Photorhabdus luminescens. AB - A proteolytic enzyme, Php-B ( Photorhabdus protease B), was purified from the entomopathogenic bacterium, Photorhabdus luminescens. The enzyme is intracellular, and its molecular mass is 74 kDa. Tested on various peptide and oligopeptide substrates, Php-B hydrolysed only oligopeptides, with significant activity against bradykinin and a 2-furylacryloyl-blocked peptide, Fua-LGPA (2 furylacryloyl-Leu-Gly-Pro-Ala; kcat=3.6x10(2) s(-1), K(m)=5.8x10(-5) M(-1), pH optimum approx. 7.0). The p K(a1) and the p K(a2) values of the enzyme activity (6.1 and 7.9 respectively), as well as experiments with enzyme inhibitors and bivalent metal ions, suggest that the activity of Php-B is dependent on histidine and cysteine residues, but not on serine residues, and that it is a metalloprotease, which most probably uses Zn2+ as a catalytic ion. The enzyme's ability to cleave oligopeptides that contain a sequence similar to collagen repeat (-Pro-Xaa-Gly-), bradykinin and Fua-LGPA (a synthetic substrate for bacterial collagenases and collagen peptidases), but not native collagens (types I and IV) or denatured collagen (gelatin), indicates that Php-B is probably a collagen peptidase, the first enzyme of this type to be identified in an insect pathogen, that might have a role in the nutrition of P. luminescens by degrading small collagen fragments. For the determination of enzyme kinetic constants, we fitted a numerically integrated Michaelis-Menten model to the experimental progress curves. Since this approach has not been used before in the characterization of proteases that are specific for the P1'-P4' substrate sites (e.g. collagenolytic enzymes), we present a comparison of this method with more conventional ones. The results confirm the reliability of the numerical integration method in the kinetic analysis of collagen-peptide-hydrolysing enzymes. PMID- 14744264 TI - Hypothermia--in the operating room and beyond. PMID- 14744265 TI - Characteristics of demand oxygen delivery systems: maximum output and setting recommendations. AB - BACKGROUND: Demand oxygen delivery systems (DODS) allot oxygen by interrupting the oxygen flow during exhalation, when it would mostly be wasted. Because DODS conserve oxygen by various methods, there are important performance differences between DODS. We studied certain performance factors that have not previously been carefully examined. METHODS: A bench model was constructed to simulate a nose, airway, and alveolar chamber. A breathing simulator generated 4 respiratory patterns, at frequencies of 15, 20, 25, and 30 breaths/min. Eighteen models of DODS were tested at 4 settings, each up to the maximum output, and compared to continuous-flow oxygen. The variable of interest was the fraction of inspired oxygen (F(I)O(2)) in the alveolar chamber, which was measured for each condition. RESULTS: The DODS differed from continuous-flow oxygen, delivering 0.5-2.1 times (mean = 1.13 times) the F(I)O(2) increase at similar settings. During maximum output the DODS showed a wide range of F(I)O(2), from 0.27 to 0.46. There was a direct relationship between volume output per pulse in the first 0.6 s of inhalation and the delivered F(I)O(2). CONCLUSIONS: DODS settings were not equivalent to continuous-flow oxygen in a bench model assessment; with equivalent settings the DODS tended to deliver greater F(I)O(2) than did continuous-flow oxygen. The maximum output capacity differed markedly among the DODS, and the user should know the device's capacity. A volume-referenced setting system for DODS should be adopted that would allow more predictable oxygen prescription and delivery via DODS. PMID- 14744266 TI - Laboratory evaluation of 4 brands of endotracheal tube cuff inflator. AB - INTRODUCTION: Routine measurement of endotracheal tube (ETT) cuff pressure is a standard in respiratory care, and several devices are available for measuring ETT cuff pressure. Yet an informed choice in the buying process is hindered by the present paucity of unbiased, comparative data. METHODS: Four brands of cuff inflator were tested: Posey Cufflator, DHD Cuff-Mate 2, Rusch Endotest, and SIMS Portex Cuff Pressure Indicator. Ten randomly selected 8.0-mm-inner-diameter ETTs were modified and tested in a trachea model. The cuffs were gradually inflated and deflated. After each sequential change in cuff volume, cuff pressure measurements were simultaneously recorded with the cuff inflator and with a calibration analyzer. These data were compared using limits-of-agreement analysis. Then, with each of the 10 ETTs, each cuff inflator was used to measure 3 known (ie, measured with the calibration analyzer) cuff pressures: 20, 40, and 60 cm H(2)O. Cuff pressure measurements were averaged, by brand, and compared to the respective baseline cuff pressure. Finally, using the 10 ETTs and trachea model, the ETT cuffs were inflated, in 0.25-mL increments, using only a syringe and the calibration analyzer. The cuff pressure and cuff volume data from that procedure were plotted and the best-fit regression line was determined. RESULTS: There were differences in bias and precision among the tested cuff inflators. The Cuff-Mate 2 had the smallest bias and best precision. None of the cuff inflator brands accurately measured cuff pressure. In each case the Cuff-Mate 2 measured cuff pressures closest to actual. The Cuff-Mate 2 contains about half the compressible volume of that in the Endotest and Cufflator and < 20% of that in the Cuff Pressure Indicator. Regarding the relationship between cuff pressure and intracuff volume, the best-fit linear regression equation was: cuff volume = 0.05 x CP - 0.39 (r(2) = 0.96). CONCLUSIONS: The 4 cuff inflators tested differ in bias and precision and none of the devices accurately measure cuff pressure. Cuff inflator manufacturers should design an accurate yet reasonably priced device to inflate ETT cuffs, and ideally that device should allow cuff-pressure checks without decreasing cuff pressure. In the meanwhile clinicians may opt to use my proposed cuff-pressure measurement technique, which minimizes the loss of cuff pressure during cuff-pressure checks and provides more accurate cuff-pressure measurements. PMID- 14744267 TI - Performance comparison of nebulizer designs: constant-output, breath-enhanced, and dosimetric. AB - INTRODUCTION: Design differences among pneumatically powered, small-volume nebulizers affect drug disposition (percentage of the dose delivered to the patient, lost to deposition in the equipment, and lost via exhalation to ambient air) and thus affect drug availability and efficacy. OBJECTIVE: Evaluate in vitro the dose disposition with 5 nebulizer models, of 3 types (constant-output, breath enhanced, and dosimetric), using simulated normal, adult breathing. METHODS: We compared 5 nebulizer models: 2 constant-output (Misty-Neb and SideStream), 1 breath-enhanced (Pari LCD), and 2 dosimetric (Circulaire and AeroEclipse). Each nebulizer was filled with a 3-mL unit-dose of albuterol sulfate and powered by oxygen at 8 L/min. The nebulizers were connected to an induction throat, connected to a breathing simulator. We measured (1) inhaled drug (subdivided into mass deposited in the induction throat and mass deposited in the filter at the distal end of the induction throat), (2) exhaled drug (lost to the ambient air), (3) drug lost to deposition in the apparatus, and (4) drug left in the unit-dose bottle. The duration of nebulization (until sputter) was measured with a stopwatch. All drug amounts were analyzed via spectrophotometry and expressed as a percentage of the total dose. RESULTS: The mean +/- SD inhaled drug percentages were: Misty-Neb 17.2 +/- 0.4%, SideStream 15.8 +/- 2.8%, Pari LCD 15.2 +/- 4.2%, Circulaire 8.7 +/- 1.0%, and AeroEclipse 38.7 +/- 1.3%. The mean +/- SD percentages of drug lost to the ambient air were: Misty-Neb 26.8 +/- 0.7%, SideStream 17.3 +/- 0.4%, Pari LCD 18.3 +/- 0.8%, Circulaire 12.3 +/- 0.8%, and AeroEclipse 6.6 +/- 3.3%. The mean +/- SD percentages of drug lost to deposition in the apparatus were: Misty-Neb 52.3 +/- 0.6%, SideStream 63.4 +/- 3.0%, Pari LCD 62.5 +/- 4.0%, Circulaire 75.8 +/- 0.5%, and AeroEclipse 51.0 +/- 2.1%. Duration of nebulization was shortest with the Circulaire and longest with the AeroEclipse (p < 0.05 via 1-way analysis of variance). CONCLUSIONS: The nebulizers we tested differ significantly in overall drug disposition. The dosimetric AeroEclipse provided the largest inhaled drug mass and the lowest loss to ambient air, with the test conditions we used. method. PMID- 14744268 TI - False-positive sputum cytology in a case of pulmonary infarction. AB - Sputum cytology is an important diagnostic tool in pulmonary medicine, but it can yield a false-positive diagnosis of malignancy. We describe such a case, which involved a 70-year-old man who presented with chest pain, hemoptysis, and bilateral pulmonary infiltrates. In the initial evaluation of hemoptysis, multiple sputum samples demonstrated cytological abnormalities consistent with adenocarcinoma, but bronchoscopy found no evidence of malignancy. He was ultimately found to have pulmonary thromboembolic disease with infarction. Follow up radiographs showed resolution of the pulmonary infarcts, and the absence of malignancy was proven during postmortem examination. Pulmonary infarction is one of many diseases that can produce sputum cytological findings falsely indicative of malignancy. PMID- 14744269 TI - Severe respiratory muscle weakness related to long-term colchicine therapy. AB - We report a case of colchicine-induced myopathy, in which the initial presenting and predominant clinical feature was respiratory muscle dysfunction; there was also chronic renal failure and electromyographically-measured myopathy. Discontinuation of colchicine led to marked improvement. Colchicine discontinuation was the only therapy performed (other medications were unchanged), and within 3 weeks the patient had regained motor function and resumed daily activities. Myopathy from primary biliary cirrhosis was ruled out. In contrast to acute colchicine intoxication, chronic colchicine toxicity is related to prolonged use rather than colchicine serum level, so colchicine serum level was not measured and did not affect the decision to discontinue colchicine. Although the diagnosis was not confirmed by muscle biopsy, we believe the typical presentation and the rapid improvement after withdrawing colchicine confirm the diagnosis. We conclude that long-term colchicine therapy, especially in the setting of chronic renal failure, can produce symptomatic respiratory muscle weakness. PMID- 14744270 TI - The evaluation and management of accidental hypothermia. AB - Accidental hypothermia is defined as an unintentional decrease in core body temperature to below 35 degrees C. Hypothermia causes hundreds of deaths in the United States annually. Victims of accidental hypothermia present year-round and in all climates with a potentially confusing array of signs and symptoms, but increasing severity of hypothermia produces a predictable pattern of systemic organ dysfunction and associated clinical manifestations. The management of hypothermic patients differs in several important respects from that of euthermic patients, so advance knowledge about hypothermia is prerequisite to optimal management. The paucity of randomized clinical trials with hypothermic patients precludes creation of evidence-based treatment guidelines, but a clinically sound management strategy, tailored to individual patient characteristics and institutional expertise and resources, can nonetheless be gleaned from the literature. This article reviews the epidemiology, pathophysiology, clinical presentation, and treatment of accidental hypothermia. Initial evaluation and stabilization, selection of a rewarming strategy, and criteria for withholding or withdrawing support are discussed. PMID- 14744271 TI - Survey of United States neurosurgical residency program directors. AB - OBJECTIVE: The field of neurosurgery in the United States faces many challenges. Neurosurgical program directors in the United States represent a logical source for inquiries about manpower issues, the training process, and Residency Review Committee (RRC) oversight. METHODS: Ninety-one active residency program directors were sent an anonymous 31-question survey. The respondents were given the option of adding additional comments. The questions were designed to address issues related to manpower, the training process, and RRC governance. Sixty-one responses were returned before an email reminder and 11 after the reminder (a total response rate of 79%). The data were entered into a database, and a descriptive analysis, with frequency distribution, was performed. RESULTS: The purpose of this review was to gain a preliminary understanding of the perceptions of program directors regarding the neurosurgical training process, the RRC, the oversight process, and projected manpower needs. A 79% response rate is high for a mail survey and likely reflects heightened concern and interest in such issues. The survey responses indicate general satisfaction with the role and governance of the RRC, significantly divergent perceptions of resident output and available positions, and serious concerns regarding the current training process. CONCLUSION: This survey suggests that a broader discussion of resident training issues would be valuable, perhaps using validated survey instruments. PMID- 14744272 TI - Methods and design considerations for randomized clinical trials evaluating surgical or endovascular treatments for cerebrovascular diseases. AB - OBJECTIVE: The results of clinical trials affect the practice of surgery and endovascular therapy for cerebrovascular diseases. The purpose of this report is to review the basic components of the designs and methods for randomized clinical trials and to describe the influence of those components on the interpretation of trial results. METHODS: The goal of an optimal clinical trial of a new procedure is to provide the most objective and rigorous evaluation of the safety and effectiveness of that procedure. Anything in the design, performance, or analysis that impairs such an assessment decreases the ability of the trial to achieve its goal and answer the research question. To highlight the components of a clinical trial, this report uses examples of Phase III clinical trials that have influenced the practice of cerebrovascular surgery and endovascular therapy in the past three decades, including the International Cooperative Study of Extracranial/Intracranial Arterial Anastomosis, the North American Symptomatic Carotid Endarterectomy Trial, the Asymptomatic Carotid Atherosclerosis Study, the Prolyse in Acute Cerebral Thromboembolism II study, and the International Subarachnoid Aneurysm Trial. RESULTS: The research question (objective) of the trial must be clearly defined, with an objective measure of efficacy and a specified quantitative difference to define the superiority of one intervention over another, in a relatively homogeneous patient population. Allocation concealment, randomization with or without stratification, and blinding (or masking) are important strategies to prevent differences in the study populations that could adversely affect the conclusions of the study. The primary end point must correspond to the specific aims of the trial. It should be objectively defined, quantifiable, reliable, and reproducible. Commonly defined end points in surgical trials include changes from baseline illness or disease severity scores, morbidity and mortality rates, and relative risks of reaching an end point with time. The statistical methods used for interim and final analyses are important. The effects of dropouts, crossovers, and missing data should be understood in the context of the final analysis. Additional concepts, such as intention-to-treat analysis and use of actual versus predicted outcomes, are important with respect to interpretation of the final results of the study. CONCLUSION: The neurosurgical and neuroendovascular communities are currently planning or conducting several clinical trials to evaluate new procedures for the treatment of cerebrovascular diseases. It is hoped that a better understanding of the components of clinical trials will facilitate the design and implementation of effective studies. PMID- 14744273 TI - Endovascular coil occlusion of 1811 intracranial aneurysms: early angiographic and clinical results. AB - OBJECTIVE: We sought to evaluate the early angiographic and clinical outcomes of the first session of endovascular coil occlusion of a large number of patients with intracranial aneurysms treated by five neurointerventionalists during a decade at a single center. METHODS: We performed retrospective analyses of pre- and postprocedural angiographic studies and early clinical outcomes. Enrolled patients underwent endovascular treatment of intracranial aneurysms with detachable coils. RESULTS: A total of 1811 aneurysms in 1579 patients were treated with coil occlusion. Of these, 90 to 100% occlusion of 86.5% of the aneurysms was achieved. In 82.3% of the procedures, no complications occurred. The clinical outcome profile at primary discharge according to the Glasgow Outcome Scale was as follows: Grade V, 74.6%; Grade IV, 6.7%; Grade III, 11.1%; Grade II, 3.1%; and Grade I, 4.5%. In patients with large aneurysms with wide necks, a lower occlusion rate and an increased complication rate were encountered. The use of three-dimensional and fibered coils resulted in higher occlusion rates. Balloon remodeling and stent deployment increased the complication rate. Previous aneurysm rupture, procedural complications, and vasospasm correlated with poor outcome. Of the patients in poor grade after aneurysm rupture, 42% recovered to Glasgow Outcome Scale Grade IV or V, as opposed to 90% of patients who were treated for unruptured aneurysms. The ischemic complication rate was 9%, and the hemorrhagic complication rate was 3%. The early procedural morbidity rate was 5.3%, and the procedural mortality rate was 1.5%. The management mortality rate was 4.4%. CONCLUSION: These data confirm the safety and efficacy of endovascular coil occlusion for patients with intracranial aneurysms. PMID- 14744274 TI - Microsurgical treatment of basilar apex aneurysms: perioperative and long-term clinical outcome. AB - OBJECTIVE: We sought to analyze the perioperative and long-term clinical outcome data for patients with microsurgically treated basilar apex aneurysms. METHODS: We identified 98 consecutively treated basilar apex aneurysms in patients prospectively enrolled in a cerebral aneurysm database. RESULTS: Fifty patients presented with subarachnoid hemorrhage, and 19 aneurysms were giant. Eighty-four of 98 aneurysms were directly clipped. Surgical morbidity was 19.4% for the entire cohort and 8.8% for the unruptured, nongiant subgroup. The most common complication resulting in long-term morbidity was perforator injury. Sixty-seven percent of patients with clipped aneurysms were independent at discharge; this fraction increased to 79.0% at the 3-month follow-up examination. Good long-term outcomes (modified Rankin Scale score < or =2) were achieved in 56 (70%) of 80 cases. The mean Barthel Index of surviving patients was 95.8 +/- 15.0 (median = 100, n = 66). Patients with unruptured, nongiant lesions fared considerably better than patients in other cohorts. Ninety-three percent of this subgroup was independent at discharge; this fraction increased to 100% at the 3-month follow up examination (n = 27). In univariate analyses, poor clinical grade, giant aneurysm size, major operative complications, and operations performed early in the series were associated with worse outcomes. In the multivariate analysis, unruptured giant aneurysm status was found to confer a tremendous risk for poor outcome (risk ratio, 80.0; 95% confidence interval, 8.0-800.7; P < 0.01). Surviving patients were observed for a mean clinical follow-up period of 7.4 +/- 3.7 years. The annual rate of postoperative subarachnoid hemorrhage was 0.18% for all clipped aneurysms and 0% for completely clipped lesions. CONCLUSION: In comparison to data from the existing literature regarding Guglielmi detachable coil embolization of basilar apex aneurysms, the data presented suggest that surgical clipping should be an important component of a multimodality approach to the treatment of patients with basilar apex aneurysms. PMID- 14744275 TI - Surgically treated aneurysms previously coiled: lessons learned. AB - OBJECTIVE: Intravascular coil embolization of cerebral aneurysms has proved to be a safe and effective treatment in certain patient groups; however, this treatment is relatively new, and the long-term outcomes are unknown. One of the known complications is refilling of the aneurysm dome, which is seen in follow-up studies. This patient population poses unique technical difficulties for the neurosurgeon. We present a series of 18 patients who underwent surgery for residual aneurysms after coil remobilization. METHODS: During a 5-year period, we performed surgery in 18 patients who had previously undergone coil embolization for their aneurysms. Of these aneurysms, four were in the anterior communicating artery, five were in the posterior communicating artery, three were in the internal carotid artery, three were in the posteroinferior cerebellar artery, and three were in the middle cerebral artery. One patient presented with rupture, one presented with acute IIIrd cranial nerve palsy, and the rest of the aneurysms were found on routine follow-up angiograms. Fifteen aneurysms were clipped, and in three patients, they were wrapped because the clip could not be placed adequately. RESULTS: There were no major complications in any of the patients, and all had uneventful recoveries. The presence of coils in the aneurysm dome and/or neck made clipping and exposure of the aneurysm neck difficult, resulting in incomplete neck obliteration in three patients. CONCLUSION: Operative clipping after previous coil embolization in aneurysms poses a unique problem for neurosurgeons. With the increasing use of coil embolization, this patient population will undoubtedly increase. The neurosurgeon should be aware of the difficulties and pitfalls encountered in these patients. PMID- 14744276 TI - Safety of high-dose intravenous eptifibatide as an adjunct to internal carotid artery angioplasty and stent placement: a prospective registry. AB - OBJECTIVE: Eptifibatide, a competitive platelet glycoprotein IIb-IIIa receptor inhibitor with high selectivity for platelet glycoprotein IIb-IIIa receptors and a short half-life, has been shown to reduce the risk of ischemic events associated with coronary interventions, particularly when used in high doses. However, its role in conjunction with neurointerventional procedures needs to be determined. We report the results of an open-label prospective registry to evaluate the safety (in terms of avoiding hemorrhagic complications) and effectiveness (in terms of preventing ischemic complications such as stroke) of administering high-dose eptifibatide during internal carotid artery angioplasty and stent placement (CAS) for extracranial carotid artery stenosis. METHODS: After femoral artery access was established and intravenous heparin (30 U/kg bolus) was administered, each patient was administered intravenous eptifibatide (two 180-microg/kg single-dose boluses before CAS, then a 2.0-microg/kg/min infusion for 20-24 hours thereafter). The primary end point was the 30-day composite occurrence of death, cerebral infarction, and unplanned or urgent endovascular or surgical intervention. The primary safety end point was bleeding, for which complications were classified according to the Thrombolysis in Myocardial Infarction scheme as major (hemoglobin decrease of more than 5 g/dl), minor (hemoglobin decrease of 3-5 g/dl), or insignificant. Platelet aggregation was measured in 13 consecutive patients with a rapid platelet-function analyzer. RESULTS: Twenty-six patients (mean age, 68.1 +/- 9.4 yr; 16 men) underwent treatment. The infusion and the CAS procedure were discontinued in one patient who developed angioneurotic edema after being administered intravenous heparin and the first bolus dose of eptifibatide. Among the 25 patients who underwent the procedure, no intracerebral hemorrhages and one minor ischemic stroke occurred during the 1-month follow-up period. The minor stroke was observed on postprocedure Day 7 in a patient for whom antiplatelet therapy was discontinued before a coronary artery bypass graft operation was performed. Another patient was discharged after an uncomplicated hospitalization but died as a result of urinary sepsis 12 days after CAS. One episode of major bleeding from the femoral insertion site required surgical repair and blood transfusions. Minor bleeding occurred in one patient. Platelet aggregation measurements obtained in 13 patients revealed a high degree (mean, 96%; range, 86-100%) of platelet inhibition after the administration of the second bolus dose of intravenous eptifibatide. CONCLUSION: High-dose eptifibatide administered as an adjunct to CAS seems to be safe. Further studies are required to analyze its effectiveness and role in neurointerventional procedures. PMID- 14744277 TI - Carotid angioplasty and stenting versus carotid endarterectomy for treatment of asymptomatic carotid stenosis: a randomized trial in a community hospital. AB - OBJECTIVE: Carotid endarterectomy (CEA) is effective in reducing the risk of stroke in individuals with more than 60% carotid stenosis. Carotid angioplasty and stenting (CAS) has been proffered as effective and used in treating individuals with asymptomatic carotid stenosis despite the absence of proven clinical equivalency. This randomized trial was designed to explore the hypothesis that CAS is equivalent to CEA for treating asymptomatic carotid stenosis. METHODS: A total of 85 individuals presenting with asymptomatic carotid stenosis of more than 80% were selected randomly for CAS or CEA and followed up for 48 months. RESULTS: Stenosis decreased to an average of 5% after CAS. The patency of the reconstructed artery remained satisfactory regardless of the technique, as determined by carotid ultrasonography. No major complications such as cerebral ischemia or death occurred. Procedural complications associated with CAS (n = 5) were hypotension and/or bradycardia; those concomitant with CEA (n = 3) were cervical nerve injury or complications related to general anesthesia (n = 4). Both procedures were well tolerated in the context of pain and discomfort. Hospital stay was similar in the two groups (mean, 1.1 versus 1.2 d). The occurrence of complications associated with CAS or CEA prolonged hospitalization by 3 days (mean, 4.0 versus 4.5 d). Return to full activity was achieved within 1 week by more than 85% of patients; all returned to their usual lifestyle by 2 weeks. Although hospital charges were slightly higher for CAS, costs were similar. CONCLUSION: CAS and CEA may be equally effective and safe in treating individuals with asymptomatic carotid stenosis. PMID- 14744278 TI - Does intracisternal thrombolysis prevent vasospasm after aneurysmal subarachnoid hemorrhage? A meta-analysis. AB - OBJECTIVE: Despite existing strategies for the treatment of vasospasm after aneurysmal subarachnoid hemorrhage, vasospasm remains a persistent contributor to death and disability. The intracisternal application of thrombolytic agents to dissolve subarachnoid clot has been advocated. The goal of this analysis was to assess the currently available evidence regarding the effectiveness of this treatment. METHODS: We conducted a systematic review of the published literature; all controlled trials were included. The outcomes of interest were delayed ischemic neurological deficits, poor Glasgow Outcome Scale scores, and death. A formal meta-analysis was performed with a random-effects model. RESULTS: The search revealed nine trials or trial subgroups (only one of which was randomized), with a total enrollment of 652 patients. Pooled results demonstrated beneficial effects of treatment, with absolute risk reductions of 14.4% (95% confidence interval, 6.5-22.5%; P < 0.001) for delayed ischemic neurological deficits, 9.5% (95% confidence interval, 4.2-14.8%; P < 0.01) for poor Glasgow Outcome Scale scores, and 4.5% (95% confidence interval, 1.5-7.5%; P < 0.05) for death. Regression analysis revealed that treatment effects did not significantly differ among the studies on the basis of the type of thrombolytic agent used (tissue plasminogen activator versus urokinase) or the method of administration (intraoperative versus postoperative) (P > 0.10). Studies that enrolled only patients at high risk for vasospasm seemed to demonstrate greater treatment effects. CONCLUSION: The meta-analysis suggests a clinically relevant and statistically significant beneficial effect of intracisternal thrombolysis. However, the results of the analysis are limited by the predominance of nonrandomized studies. Further randomized, blinded, placebo-controlled trials of high-risk patients would be justified. PMID- 14744279 TI - Detection of intracranial aneurysms with two-dimensional and three-dimensional multislice helical computed tomographic angiography. AB - OBJECTIVE: Computed tomographic angiography (CTA) has become a diagnostic method for the detection of intracranial aneurysms in cases of subarachnoid bleeding. We sought to evaluate the detection of aneurysms with CTA with a novel multislice helical computed tomographic scanner. METHODS: Prospectively, 179 patients underwent multislice CTA, followed by digital subtraction angiography (DSA) of both carotid arteries with or without the posterior circulation, DSA of one carotid artery with or without the posterior circulation, or DSA of the posterior circulation alone. The total number of carotid arteries studied was 298, and the number of vertebrobasilar arteries studied was 124. RESULTS: Of 178 aneurysms verified with DSA or intraoperatively, CTA failed to detect 7 aneurysms of 1 to 2 mm and 1 partially thrombosed, 4-mm aneurysm. The sensitivity and specificity of CTA for aneurysm detection were 0.96 and 0.97, respectively. CONCLUSION: The first generation of multislice computed tomographic technology does not improve CTA to surpass DSA for the detection of small aneurysms of 1 to 2 mm. In practice, however, CTA is superior as a fast noninvasive method without complications. PMID- 14744280 TI - Healing process for cerebral dissecting aneurysms presenting with subarachnoid hemorrhage. AB - OBJECTIVE: This was a pathological study to investigate the healing process for cerebral dissecting aneurysms presenting with subarachnoid hemorrhage (SAH). METHODS: Thirteen dissecting aneurysms that presented with SAH were obtained from 13 patients. Nine aneurysms arose from the vertebral artery, two arose from the anterior cerebral artery, one arose from the internal carotid artery, and one arose from the superior cerebellar artery. Eight aneurysm specimens were collected during autopsy and five were resected during surgery (trapping with or without bypass). The period between the onset of SAH and the time of specimen collection ranged from 6 hours to 35 days. All 13 aneurysms were pathologically examined with immunohistochemical staining, with a focus on the chronological healing process after SAH. RESULTS: All dissecting aneurysms were generated with sudden widespread disruption of the internal elastic lamina and media. The healing process occurred with neointimal proliferation. The neointima, consisting mainly of newly synthesized smooth muscle cells and collagen fibers, extended from the disrupted ends of the media proper forward to the ruptured portion. CONCLUSION: It is assumed that the healing process, with neointimal proliferation, begins after 1 week and may not be complete even after 1 month, depending on the extent of the wall injury. PMID- 14744281 TI - Correlation between promoter hypermethylation of the O6-methylguanine deoxyribonucleic acid methyltransferase gene and prognosis in patients with high grade astrocytic tumors treated with surgery, radiotherapy, and 1-(4-amino-2 methyl-5-pyrimidinyl)methyl-3-(2-chloroethyl)-3-nitrosourea-based chemotherapy. AB - OBJECTIVE: O(6)-Methylguanine-deoxyribonucleic acid methyltransferase (MGMT) is a deoxyribonucleic acid repair protein associated with the chemoresistance of chloroethylnitrosoureas. We investigated whether MGMT promoter hypermethylation is associated with prognosis in patients with high-grade astrocytic tumors treated uniformly with surgery, radiotherapy, and 1-(4-amino-2-methyl-5 pyrimidinyl)methyl-3-(2-chloroethyl)-3-nitrosourea (ACNU)-based chemotherapy. METHODS: Using the methylation-specific polymerase chain reaction, we assayed promoter hypermethylation of the MGMT gene in tumor deoxyribonucleic acid from 116 adult patients with supratentorial high-grade astrocytic tumors (42 anaplastic astrocytomas [AAs] and 74 glioblastomas multiforme [GBMs]). The Cox proportional hazards model was used in forward stepwise regression to assess the relative role of prognostic factors (i.e., age at surgery, sex, Karnofsky Performance Scale score, extent of surgical resection, methylation status of the MGMT promoter, and association between MGMT promoter methylation and survival). RESULTS: MGMT promoter hypermethylation was confirmed in 19 (45.2%) of 42 AA patients and 33 (44.6%) of 74 GBM patients. It was significantly associated with both longer overall and progression-free survival time in AA but not GBM patients. CONCLUSION: Our results demonstrate that MGMT promoter hypermethylation is associated with longer survival time in patients with AA who were treated with surgery, radiotherapy, and ACNU-based chemotherapy but not in patients with GBM. PMID- 14744282 TI - Depression in patients with high-grade glioma: results of the Glioma Outcomes Project. AB - OBJECTIVE: To study the incidence of depression among patients undergoing surgery for high-grade glioma, document factors associated with the presence of depression, and examine the relationship between depression and patient outcome. METHODS: Physician and patient reports of depression were analyzed immediately postoperatively and again 3 and 6 months after surgery for high-grade glioma. Physician-reported depression was defined according to the Diagnostic and Statistical Manual of Mental Disorders, ed 4. Patient self-assessment of depression was based on responses to questions contained in two validated functional status surveys. Concordance of physician- and patient-reported depression was examined, along with the extent of use of pharmacological treatment for depression. Additional outcomes examined included quality of life, survival, patient satisfaction, and posttreatment complications. RESULTS: Data from 598 patients were analyzed. In the early postoperative period, physicians reported depression in 15% of patients, whereas 93% of patients reported symptoms consistent with depression. The incidence of patient self-reported depression remained similar at 3- and 6-month follow-up, whereas physician reported depression increased from 15% in the early postoperative period to 22% at both 3- and 6-month follow-up. Concordance between physician recognition of depression and treatment of depression was low initially (33%) and increased at 3 and 6 months (51 and 60%, respectively). As compared with patients who were not depressed, survival was shorter and complications were more common among depressed patients. CONCLUSION: Symptoms of depression were common immediately after surgery for glioma, and they increased throughout the 6-month period after surgery. These findings support the hypothesis that clinically important depression is a common complication in patients with high-grade glioma. Concordance between physician recognition of depression and self-reports of depression by patients was low. Concordance between physician recognition of depression and initiation of pharmacological antidepressant therapy was fair in the early postoperative period and improved somewhat over the subsequent 6-month period; however, within the 6-month period after surgery for glioma, antidepressant therapy was provided for only 60% of patients in whom the physician recognized depressive symptoms and in only 15% of patients who self reported symptoms of depression. Findings from this observational study suggest the need for a controlled trial that is designed to test the hypothesis that more attention to the identification of postoperative depression and aggressive treatment of depressive symptoms can improve the quality of life and survival of patients after surgery for high-grade glioma. PMID- 14744283 TI - Transforaminal lumbar interbody fusion: surgical technique and results in 24 patients. AB - OBJECTIVE: The advantage of anterior column support and fusion in addition to pedicle fixation in patients with degenerative spinal disorders has become increasingly clear. With the increase in popularity of this treatment, a variety of techniques have been used to achieve the goal of anterior column support, fusion, and segmental instrumentation. Posterior lumbar interbody fusion has been used since the late 1940s in the treatment of degenerative lumbar spine. We evaluated a modification to posterior lumbar interbody fusion called transforaminal lumbar interbody fusion (TLIF). METHODS: A retrospective analysis was performed on 24 patients (9 women, 15 men) who underwent TLIF. The approach involved a unilateral laminectomy and inferior facetectomy at the level of fusion. The interbody fusion was achieved from this unilateral approach by performing discectomy, arthrodesis, and insertion of one or two titanium cages packed with autologous bone. The average age of the patients in this study was 42.6 +/- 12.5 years. Five patients were smokers. Five cases were related to workmen's compensation. Seventeen patients' original symptoms were a combination of low back pain and radiculopathy. Ten patients had had a previous spine operation. RESULTS: Eleven patients had L4-S1 TLIFs. The rest of the patients had a single-level TLIF (L2-S1). Average intensive care unit and floor days were 1.1 +/- 1.0 and 5.8 +/- 2.2 days, respectively. The number of days to ambulation was 2.8 +/- 1.6 days. There were a total of six self-limited complications in 24 patients (including one transient neurological complication). The average follow up time was 16.9 +/- 9.1 months. Twenty-two patients had solid fusions. A modified Prolo scale (4 worst, 20 best) was used to evaluate the clinical outcome. The average score was 16.1 +/- 4.1. CONCLUSION: TLIF is a reliable and safe technique for interbody support that can be performed with excellent clinical outcome. In the authors' experience, TLIF offers excellent exposure with minimal risk. This applies particularly in cases of repeat spine surgery, in which the presence of scar tissue makes traditional posterior lumbar interbody fusion techniques difficult or impossible. In addition, TLIF seems to be a viable alternative to anteroposterior circumferential fusion and/or anterior lumbar interbody fusion. PMID- 14744284 TI - The value of magnetic resonance imaging in the evaluation of fatty filum terminale. AB - OBJECTIVE: To determine whether there are magnetic resonance imaging (MRI) characteristics of fatty fila that are correlated with neurological deficits, especially in the presence of a normal-level conus medullaris. METHODS: Lumbosacral MRI scans were reviewed for patients with fatty fila who were treated at Duke University Medical Center during a 5-year period. The patients were divided into three groups. Group I patients (n = 5) had fatty fila that were incidentally detected during evaluations for metastases or infections. Group II patients (n = 16) exhibited isolated low back pain but were in neurologically intact condition. Group III patients (n = 15) exhibited neurological impairments consistent with distal spinal cord dysfunction. Several characteristics were measured on the MRI scans, including the location of the conus medullaris, the filum thickness, and the distance of fat from the conus. These results were assessed for statistically significant correlation with the presence of clinical symptoms. RESULTS: The majority of patients in all three groups demonstrated the normal conus position (L2 or above) and thickened fila. The distance of fat from the conus was the only parameter that demonstrated a statistically significant difference among the groups. CONCLUSION: The following findings were noted: 1) patients were likely to exhibit neurological deficits at a younger age (<22 yr in Group III versus 47 yr in Groups I and II); 2) a conus level below L2 was associated with neurological deficits (Group III); 3) filum thickness was not correlated with clinical presentation; 4) fat in the filum within 13 mm of the conus medullaris was most predictive of neurological deficits (Group III). PMID- 14744285 TI - Functional anatomy of the human cochlear nerve and its role in microvascular decompressions for tinnitus. AB - OBJECTIVE: The functional anatomy (i.e., tonotopy) of the human cochlear nerve is unknown. A better understanding of the tonotopy of the central nervous system segment of the cochlear nerve and of the pathophysiology of tinnitus might help to ameliorate the disappointing results obtained with microvascular decompressions in patients with tinnitus. METHODS: We assume that vascular compression of the cochlear nerve can induce a frequency-specific form of hearing loss and that when the nerve is successfully decompressed, this hearing loss can recuperate. Thirty-one patients underwent a microvascular decompression of the vestibulocochlear nerve for vertigo or tinnitus. Preoperative audiograms were subtracted from postoperative audiograms, regardless of the surgical result with regard to the tinnitus and vertigo, because the hearing improvement could be the only sign of the vascular compression. The frequency of maximal improvement was then correlated to the site of vascular compression. A tonotopy of the cochlear nerve was thus obtained. RESULTS: A total of 18 correlations can be made between the site of compression and postoperative maximal hearing improvement frequency when 5-dB hearing improvement is used as threshold, 13 when 10-dB improvement is used as threshold. A clear distribution can be seen, with clustering of low frequencies at the posterior and inferior side of the cochlear nerve, close to the brainstem, and close to the root exit zone of the facial nerve. High frequencies are distributed closer to the internal acoustic meatus and more superiorly along the posterior aspect of the cochlear nerve. CONCLUSION: The tonotopic organization of the cisternal segment of the cochlear nerve has an oblique rotatory structure as a result of the rotatory course of the cochlear nerve in the posterior fossa. Knowledge of this tonotopic organization of the auditory nerve in its cisternal course might benefit surgeons who perform microvascular decompression operations for the vestibulocochlear compression syndrome, especially in the treatment of unilateral severe tinnitus. PMID- 14744286 TI - Translabyrinthine approach for acoustic neuroma. AB - The translabyrinthine approach has been popularized during the past 30 years for the surgical treatment of acoustic neuromas. It serves as an alternative to the retrosigmoid approach in patients when hearing preservation is not a primary consideration. Patients with a tumor of any size may be treated by the translabyrinthine approach. The corridor of access to the cerebellopontine angle is shifted anteriorly in contrast to the retrosigmoid approach, resulting in minimized retraction of the cerebellum. Successful use of the approach relies on a number of technical nuances that are outlined in this article. PMID- 14744287 TI - Mini-open carpal tunnel decompression. AB - Carpal tunnel syndrome is the most common entrapment neuropathy, and it is caused by compression of the median nerve at the wrist. The authors describe the mini open carpal tunnel technique for surgical release of the transverse carpal ligament. The success of the procedure depends on meticulous technique with attention to certain important anatomic details and careful avoidance of injury to the palmar cutaneous nerve and the recurrent motor branch. PMID- 14744288 TI - Endoscopic exploration of the brachial plexus: technique and topographic anatomy- a study in fresh human cadavers. AB - OBJECTIVE: The indications for and timing of brachial plexus exploration in closed injuries are controversial. The time-consuming surgery proves its worth in some cases, whereas spontaneous regeneration might have been possible in others. The differentiation is difficult, because no investigational method reveals the exact morphological correlates of the nerve lesions. Minimally invasive, direct observation of the structures is a possible solution. Here we describe our surgical technique and the anatomic features of the normal brachial plexus appreciated with the endoscope. METHODS: Twenty-one brachial plexus in 11 fresh cadavers were investigated. Endoscopic exploration was performed at the supraclavicular and infraclavicular levels. The method involves insertion of an optic shaft-integrated retractor through a stab wound; retraction of landmark muscles produces a working space, into which other instruments are introduced for dissection. After completion of endoscopic surgery, open dissection was performed to verify the endoscopically identified structures and to assess iatrogenic injuries. RESULTS: The omohyoid muscle is a reliable landmark in the supraclavicular region, beneath which the suprascapular nerve can be observed. Following the suprascapular nerve proximally leads to the plexus trunks. Infraclavicular exploration first reveals the axillary artery. The plexus and its nerves are traced around this artery. The anatomic features were constant in all cases, with variations in fat accumulation depending on the corporeal constitution. We detected iatrogenic injuries to the medial circumflex humeral vessels in two cases. No nerve injuries were observed. CONCLUSION: The endoscopic technique combined with intraoperative nerve stimulation studies might provide important information on the type of morphological damage in closed brachial plexus injuries and thus might become an important tool for determination of the surgical treatment strategy. Clinical work is under way. PMID- 14744289 TI - Gene microarray analysis of human brain arteriovenous malformations. AB - OBJECTIVE: Human brain arteriovenous malformations (BAVMs) display abnormal expression of various angiogenesis-related genes and their products. We examined gene expression patterns in BAVMs by the gene microarray technique. METHODS: We analyzed BAVM and control brain samples obtained by temporal lobectomy for medically intractable seizure by Affymetrix Human Gene Set U95Av2 (Affymetrix, Inc., Santa Clara, CA). The gene microarray data were compared with new and previously published data that used conventional molecular biology techniques. RESULTS: We analyzed six BAVM and five control brain samples. From 12,625 gene probes assayed, 1781 gene probes showed differential expression between BAVMs and controls. BAVM samples had a gene expression pattern that was distinct from those of control brain samples. Increased messenger ribonucleic acid expression of vascular endothelial growth factor A was accompanied by increased expression of its protein product. A majority of the gene data was in agreement with previously published data. The gene microarray data generated a new testable hypothesis regarding integrin, and we found increased expression of integrin alphavbeta3 protein in BAVMs. CONCLUSION: The gene expression pattern of BAVMs was distinct from those of control brain samples. We verified the gene microarray data by demonstrating that increased gene expression levels for angiogenesis-related molecules were accompanied by increased levels of their protein product expression. The gene microarray technique may be a useful tool to study multiple pathways simultaneously in BAVM specimens. PMID- 14744290 TI - Experimental subarachnoid hemorrhage: cerebral blood flow and brain metabolism during the acute phase in three different models in the rat. AB - OBJECTIVE: To study the cerebral metabolism and its relationship to cerebral blood flow (CBF) acutely after subarachnoid hemorrhage (SAH). METHODS: SAH was induced in rats by endovascular perforation of the internal carotid artery, blood injection into the prechiasmatic cistern or the cisterna magna. CBF (measured by laser Doppler flowmetry), cerebral perfusion pressure, O(2) tension, and extracellular levels of glucose, lactate, and pyruvate were monitored during 90 minutes after SAH. CBF (assessed by (125)I-antipyrine autoradiography), arteriovenous O(2) difference, and cerebral metabolic rate of O(2) were calculated at 15 or 90 minutes after SAH. RESULTS: After a transient reduction, cerebral perfusion pressure normalized within 5 minutes after SAH in all groups. There was a transient global decrease in CBF after SAH: its duration depended on the severity of the hemorrhage. CBF of less than 20% of baseline was observed for at least 15 minutes in 25% and 14% of the animals after perforation and prechiasmatic SAH, respectively. In all SAH groups, O(2) tension was suddenly reduced to approximately 40% of baseline and gradually increased, reaching 70 to 90% of baseline 90 minutes after SAH. The cerebral metabolic rate of O(2) was reduced only at 15 minutes after perforation and prechiasmatic SAH, but arteriovenous O(2) difference was normal in all groups. During 30 minutes after perforation SAH, a 50% decrease in glucose and a threefold increase in lactate and pyruvate levels were observed. CONCLUSION: The data suggest that SAH induced an acute global decrease in CBF together with a depression in the cerebral metabolism. The degree of the changes was related to the severity of the hemorrhage. The metabolic derangements were not always explained by ischemic episodes. PMID- 14744291 TI - Evolution of neuroendovascular intervention: a review of advancement in device technology. AB - Neuroendovascular surgery is a rapidly evolving field. Each year, numerous improvements are made in the endovascular surgeon's armamentarium. This evolution in technology, which is occurring at a dizzying pace, addresses many of the current limitations of neuroendovascular approaches. The potential to improve the outcomes of our patients is tremendous, particularly because one of the most common and most devastating neurological disorders, ischemic stroke, remains largely untreated. This article presents several of the new technologies that are currently being investigated or are under development and have the potential to lead to major advances in endovascular approaches for the treatment of intracranial and extracranial diseases. PMID- 14744292 TI - Stereotactic navigation, Jean Talairach, and I. AB - This is not meant to be a comprehensive review of the work of Jean Talairach and his associates at the Centre Hospitalier Ste. Anne in Paris. Much more space than is available here would be required for that purpose. Talairach's contributions in the development of stereotactic instrumentation and methods, cerebral anatomy, neurophysiology, epilepsy, stereotactic biopsy and interstitial irradiation of tumors, radionuclide ablation of the pituitary gland, and surgical treatment of pain, movement disorders, and psychiatric illnesses are legend. Talairach is a creative pioneer who, from the time he arrived at Ste. Anne in 1937 at 28 years of age, has remained productive throughout his life and, even in his late eighties, continued to contribute to his field. His legacy is enormous. This work is simply meant to relate some of my personal anecdotes about Talairach and the people around him and to describe how the experience influenced my own career. PMID- 14744293 TI - A short history of the William H. Welch Medical Library. AB - The William H. Welch Medical Library was founded in 1929 with the merger of three libraries, namely, the library of the School of Medicine, the library of the School of Hygiene and Public Health, and the library of the Johns Hopkins Hospital. Engendered by the dreams of Johns Hopkins University bibliophiles, such as founding Hopkins physicians William Osler, William Stewart Halsted, Howard Kelly, and William Henry Welch, the library flourished and expanded. At the same time as the founding of the central medical library, the Institute of the History of Medicine collection was established in the Welch Library building, with William H. Welch as its first director. Innovative leadership led to many exciting projects, such as the Welch Indexing Project, which was a prototype for the National Library of Medicine's medical subject headings, and the establishment of a laboratory for research into the application of information technologies to knowledge management. The Welch Library enters the 21st century as a dynamic library committed to the continuing delivery of information services, as well as ongoing research into the development of library services for the coming 100 years. PMID- 14744294 TI - A history of Todd and his paralysis. AB - OBJECTIVE: To describe the history of Robert Bentley Todd (1809-1860) and certain of his contributions to medicine, including his original and subsequent descriptions of "epileptic hemiplegia," which came to be called "Todd's paralysis." METHODS: The author conducted a comprehensive review of English language literature, modern and historical, related to "Todd's paralysis" and examined Todd's original case histories and commentary by Todd, his contemporaries, and his successors. RESULTS: Todd held that some patients "who recover from a severe fit, or from frequently repeated fits of epilepsy, are often found to labor under hemiplegia, or other modifications of palsy." He believed that this resulted from "undue exaltation. [resulting in] a state of depression or exhaustion." Interestingly, Todd was the first to present an electrical theory of epilepsy, supported by his own animal experimentation, well before his better-known successor John Hughlings Jackson (1835-1911) (famous for his investigations of partial epilepsies and the eponymous "Jacksonian march"). CONCLUSION: Many neurologists and investigators followed Todd in acknowledging transient postictal paralysis as a distinct clinical entity. Yet whether the pathophysiology of "Todd's paralysis" is related to "neuronal exhaustion" or excessive inhibition is still controversial. PMID- 14744295 TI - Bacterial adhesion to surfactant-modified silicone surfaces. AB - OBJECTIVE: Ventricular shunt infections are a major contributor to morbidity in patients being treated for hydrocephalus. The majority of these infections are from Staphylococcus epidermidis. Prevention of bacterial adhesion to the silicone surface of a ventricular catheter could decrease shunt infections. We studied the effectiveness of a surfactant and/or 2% iodine prewash on preventing bacterial adhesion to Silastic catheter material. METHODS: In a laboratory setting, various concentrations of a surfactant, Poloxamer-188 (P188), and a bactericidal agent, iodine, were compared against a control solution in their ability to prevent bacterial adhesion of S. epidermidis to a silicone surface. Silicone wafers were soaked for 1 hour in the test solution, then inoculated and incubated with S. epidermidis for 24 hours. Bacterial counts were then obtained and compared. RESULTS: The most effective method tested in this study was 20% P188, which allowed only 3.02% bacterial adhesion compared with 22.2% bacterial adhesion in the control (P < 0.001). P188 at a 10% concentration or 20% mixed with iodine had the next most effective inhibition. Of the germicidal solutions, a 5-ppm solution of iodine was the most effective. The most ineffective method tested was 2 ppm iodine, which allowed 13.2% bacterial adhesion. CONCLUSION: Use of a surfactant and/or a germicidal will provide some protection against bacteria attaching to silicone surfaces before they are surgically implanted. The use of a surfactant soak of 20% P188 or iodine at a concentration of 5 ppm before inoculation with S. epidermidis significantly decreased the bacterial adhesion to silicone wafers. This finding has relevance to clinical practice because it highlights a simple step undertaken before implanting a ventricular catheter that could reduce the adhesion rate of the most common contaminant of these catheters. This step may become an important factor in decreasing infection rates in shunt-dependent patients. PMID- 14744296 TI - Cavernous sinus and leptomeningeal metastases arising from a squamous cell carcinoma of the face: case report. AB - OBJECTIVE AND IMPORTANCE: Invasion of trigeminal and facial perineural spaces is a recognized complication of cutaneous malignancies. Centripetal spread along the trigeminal nerve axis and into the cavernous sinus and the gasserian ganglion is rare. Metastasis to the leptomeninges and cauda equina has not been reported. We report a unique case of perineural spread and central dissemination from an epithelial squamous cell carcinoma (SCC) associated with a tumor biomarker. CLINICAL PRESENTATION: After excision of multiple cutaneous SCCs and basal cell carcinomas of the head and neck, a 70-year-old male patient developed successive, right-side, V1 and V2 trigeminal neuropathies and complete right cavernous sinus syndrome during a 5-year period. Concurrently, the right face became paralyzed. Left facial paresis developed during the latter half of this period. Two months before admission, subacute left lower-extremity radicular weakness resulted in falls. Serial magnetic resonance imaging scans obtained in the previous 4 years were unrevealing. At the time of admission, enhancing masses were found in the 1) right cavernous sinus and dura, foramina ovale and rotundum, and Meckel's cave, 2) right subtemporal region and orbital rectus muscles, and 3) cauda equina. Cerebrospinal fluid analysis demonstrated mild pleocytosis and rare carcinoma cells. INTERVENTION: Biopsy of the right cavernous sinus mass confirmed moderately differentiated, metastatic SCC. Immunohistochemical staining and fluorescence in situ hybridization revealed epidermal growth factor receptor overexpression and genomic amplification. CONCLUSION: The indolent progression of cranial nerve palsy among patients with resected cutaneous SCCs of the head and neck must raise clinical suspicion of perineural spread, even in the absence of radiological changes. Biomarkers predicting aggressive SCC behavior, illustrated here by epidermal growth factor receptor amplification and central invasion, have the potential to guide early therapy. PMID- 14744297 TI - Malignant nerve sheath tumor of the right cerebral peduncle: case report. AB - OBJECTIVE AND IMPORTANCE: Schwannomas occurring in the neuraxis are very rare. Usually, these tumors are benign. Primary malignant intracerebral nerve sheath tumors are extremely rare, with only five documented cases in the international literature. We report one case of a primary malignant intracerebral nerve sheath tumor occurring in the right cerebral peduncle of a 35-year-old man. CLINICAL PRESENTATION: Magnetic resonance imaging revealed a heterogeneous peripherally enhancing mass of the right cerebral peduncle, surrounded by a small edema. INTERVENTION: Unlike the five cases previously reported, this is the first time a stereotactic biopsy has been performed, and this is the only patient who responded to cranial radiation therapy for approximately 2 years. When the tumor recurred, a systemic chemotherapy treatment was prescribed. No positive response was seen, and the patient died 29 months after the initial diagnosis. CONCLUSION: An accurate diagnosis and planned aggressive treatment seem to be the key elements in the management of the disease. PMID- 14744299 TI - Spinal accessory schwannoma mimicking a tumor of the fourth ventricle: case report. AB - OBJECTIVE AND IMPORTANCE: Spinal accessory schwannomas unassociated with neurofibromatosis are very rare, and only 30 cases have been reported in the literature. To our knowledge, this is the first report of a spinal accessory schwannoma mimicking a tumor of the fourth ventricle. CLINICAL PRESENTATION: A 50 year-old man presented with neck pain after being involved in a motor vehicle accident. There were no neurological deficits, but a computed tomographic scan revealed a large hypodense mass with punctuate calcifications in the fourth ventricle. The tumor exhibited low intensity on the T1-weighted magnetic resonance imaging scan and high intensity on the T2-weighted scan, and it showed inhomogeneous contrast enhancement. INTERVENTION: The tumor was totally removed by a bilateral suboccipital craniectomy and C1 laminectomy. Dissection of the surgical specimen revealed that the tumor had originated from the left spinal accessory nerve. Histopathological examination confirmed the diagnosis of schwannoma. The patient experienced transient postoperative cerebellar ataxia but recovered completely. CONCLUSION: Intracisternal-type spinal accessory schwannomas sometimes mimic a tumor of the fourth ventricle. Total surgical resection can be achieved with good outcome. PMID- 14744298 TI - Trochlear nerve palsy after repeated percutaneous balloon compression for recurrent trigeminal neuralgia: case report and pathogenic considerations. AB - OBJECTIVE AND IMPORTANCE: Repeated percutaneous balloon compression for the treatment of idiopathic trigeminal neuralgia is infrequent. When a second procedure is performed, the outcome is unknown. A patient developed an isolated trochlear nerve palsy after undergoing percutaneous trigeminal ganglion balloon compression for a second time. The mechanism of diplopia and the complications associated with this technique were studied. CLINICAL PRESENTATION: The patient was a 67-year-old woman with a history of medically refractory idiopathic trigeminal neuralgia involving all three divisions of the right trigeminal nerve. INTERVENTION: Percutaneous balloon compression was performed. Despite initial total relief from pain without complications, the patient again displayed manifestations of trigeminal neuralgia 3 months after the procedure. The pain disappeared after she underwent a second balloon compression procedure, but she developed an isolated trochlear nerve palsy, which spontaneously resolved in 2 months. CONCLUSION: Isolated trochlear nerve palsy is a rare and reversible complication after percutaneous balloon compression for trigeminal neuralgia. This case illustrates that the mechanism of injury to the fourth nerve is the result of an erroneous technique: excessive penetration of the Fogarty catheter in Meckel's cave beyond the porus trigemini and compression of the cisternal segment of the trochlear nerve when the inflated balloon is pushed against the tentorium. PMID- 14744300 TI - Hypertrophic neuropathy of the cauda equina: case report. AB - OBJECTIVE AND IMPORTANCE: Hypertrophic neuropathy of the cauda equina (HNCE) is a rare form of peripheral neuropathy. The diagnosis is complicated by an insidious clinical presentation and complex radiographic images. We present a case of HNCE caused by chronic inflammatory demyelinating polyneuropathy with symptomatic improvement after decompressive lumbar laminectomy and dural expansion. CLINICAL PRESENTATION: A 54-year-old woman with a history of back pain since she was in her 20s presented with low back and radicular pain that had increased during a period of 6 months, bilateral lower-extremity weakness, and sensory loss in the right thigh. Magnetic resonance imaging of the lumbosacral spine revealed multiple, poorly enhancing mass lesions and apparent intrathecal nerve root thickening from L1 to L5. INTERVENTION: An L1-L5 decompressive laminectomy, performed with continuous somatosensory evoked potential and electromyographic monitoring, revealed multiple segmentally enlarged nerve roots. One nerve root that did not respond to high levels of stimulation was identified. This root was resected and submitted for pathological analysis. The dura was expanded with an 11-cm-long dural patch. The pathological examination revealed hypertrophic neuropathy, with extensive S-100-positive "onion bulb" formation. The patient's symptoms improved postoperatively. CONCLUSION: HNCE is a rare disorder that can cause radicular pain and lower-extremity weakness, sensory loss, and hyporeflexia. One possible cause is demyelinating polyneuropathy. Although medical management is typically effective in the treatment of demyelinating polyneuropathy, it has little effect on compressive symptoms caused by intradural nerve root enlargement. As this case demonstrates, surgical management of symptomatic radiculopathy by lumbar laminectomy is a reasonable and effective approach to the treatment of HNCE. PMID- 14744301 TI - Should children decide whether they are enrolled in nonbeneficial research? AB - The U.S. federal regulations require investigators conducting nonbeneficial research to obtain the assent of children who are capable of providing it. Unfortunately, there has been no analysis of which children are capable of assent or even what abilities ground the capacity to give assent. Why should investigators be required to obtain the positive agreement of some children, but not others, before enrolling them in research that does not offer a compensating potential for direct benefit? We argue that the scope of children's research decision making should be based on the principles of respect for autonomy and nonmaleficence. These principles imply that the threshold for assent should be fixed at 14 years of age, and a dissent requirement should be adopted for all children in the context of nonbeneficial research. PMID- 14744302 TI - Moral responsiveness in pediatric research ethics. PMID- 14744303 TI - Guiding IRBs and educating researchers. PMID- 14744304 TI - Sham surgery in randomized trials: additional requirements should be satisfied. PMID- 14744305 TI - Not just how, but whether: revisiting Hans Jonas. PMID- 14744307 TI - We need substantive criteria for decisions by children. PMID- 14744308 TI - Rethink "affirmative agreement," but abandon "assent". PMID- 14744309 TI - We should reject passive resignation in favor of requiring the assent of younger children for participation in nonbeneficial research. PMID- 14744310 TI - Knowing who you want to be when you grow up: implications for pediatric assent. PMID- 14744311 TI - Narrative portrayals of genes and human flourishing. PMID- 14744312 TI - Children's consent to research participation: social context and personal experience invalidate fixed cutoff rules. PMID- 14744313 TI - Assent and dissent in 407 research with children. PMID- 14744314 TI - The limits of altruism and arbitrary age limits. PMID- 14744315 TI - Altruism, children, and nonbeneficial research. PMID- 14744316 TI - Public mistrust: the unrecognized risk of the CDC Smallpox Vaccination Program. AB - Phase 1 of the CDC's Smallpox Vaccination Program has foundered because of a lack of volunteers. However, despite the enrollment of fewer than 10% of the projected number of hospital employees in Phase 1, and recent recommendations of two advisory groups, CDC and DHHS officials recently announced plans to expand this vaccination program. During Phase 2 of the Smallpox Vaccination Program, an additional 10 million health care and emergency workers are scheduled to receive vaccination. This paper reviews reasons why state health departments and hospitals rejected participation in Phase 1. It urges the federal government to take account of these problems before proceeding to Phase 2 and argues that the flaws in this program not only threaten bioterrorism preparedness efforts but, more importantly, might endanger trust in public health initiatives. PMID- 14744317 TI - What's altruism got to do with it? PMID- 14744318 TI - Taking children seriously: what's so important about assent? PMID- 14744319 TI - The rebirth of bioethics: extending the original formulations of Van Rensselaer Potter. AB - Van Rensselaer Potter's original concept of bioethics as a global integration of biology and values was designed to guide human survival. His attention to the creation of human knowledge and the incorporation of ecological concepts and values into medicine and health remain important, yet largely neglected, contributions deserving of further elaboration. Bioethicists should heed his warning about unsustainable progress, particularly in health care systems, and work toward changing their behaviors. Incorporating life-affirming spiritual values and extending Potter's global bioethics to a deeper bioethics seem essential. PMID- 14744320 TI - A goodness-of-fit ethic for child assent to nonbeneficial research. PMID- 14744321 TI - Context in shaping the ability of a child to assent to research. PMID- 14744322 TI - Preventing exploitation in pediatric research. PMID- 14744323 TI - Detainee ethics: terrorists as research subjects. PMID- 14744324 TI - Children under age 14 deserve more. PMID- 14744325 TI - A response to "Children in clinical research: a conflict of moral values" by Vera Hassner Sharav. PMID- 14744326 TI - From the mouths of babes: a response to "Should children decide whether they are enrolled in nonbeneficial research?" by David Wendler and Seema Shah. PMID- 14744327 TI - Why shouldn't children decide whether they are enrolled in nonbeneficial medical research? PMID- 14744328 TI - A response to commentators on "Sham surgery: an ethical analysis". PMID- 14744329 TI - Child assent revisited. PMID- 14744330 TI - A response to commentators on "Should children decide whether they are enrolled in nonbeneficial research?". PMID- 14744331 TI - Why a teenager over age 14 should be able to consent, rather than merely assent, to participation as a human subject of research. PMID- 14744332 TI - Sham surgery: an ethical analysis. AB - Surgical clinical trials have seldom used a "sham" or placebo surgical procedure as a control, owing to ethical concerns. Recently, several ethical commentators have argued that sham surgery is either inherently or presumptively unethical. In this article I contend that these arguments are mistaken and that there are no sound ethical reasons for an absolute prohibition of sham surgery in clinical trials. Reflecting on three cases of sham surgery, especially on the recently reported results of a sham-controlled trial of arthroscopic surgery for arthritis of the knee, I present an ethical analysis that focuses on the methodological rationale for use of sham surgery, risk-benefit assessment, and informed consent. PMID- 14744333 TI - Surgical research, an elusive entity. PMID- 14744334 TI - Does placebo surgery-controlled research call for new provisions to protect human research participants? PMID- 14744335 TI - If the goal is relief, what's wrong with a placebo? PMID- 14744336 TI - Sham neurosurgery in Parkinson's disease: ethical at the time. PMID- 14744337 TI - The physician's role, "sham surgery," and trust: a conflict of duties? PMID- 14744338 TI - An innovative paradigm for clinical research. PMID- 14744339 TI - Sham surgery and genuine standards of care: can the two be reconciled? PMID- 14744340 TI - Sham surgery in research: a surgeon's view. PMID- 14744341 TI - Sham surgery: to cut or not to cut--that is the ethical dilemma. PMID- 14744342 TI - The sham surgery debate and the moral complexity of risk-benefit analysis. PMID- 14744343 TI - Social aspects of sham surgeries. PMID- 14744344 TI - Is intravenous urography a prerequisite for renal shockwave lithotripsy? AB - PURPOSE: To determine whether intravenous urography (IVU) is a prerequisite for shockwave lithotripsy (SWL) of renal stones by addressing whether using non contrast-enhanced CT (NCCT) instead of IVU for delineating urinary tract anatomy is associated with post-SWL complications. PATIENTS AND METHODS: Thirty-eight patients treated by SWL (Econolith 2000) for radiopaque renal stones underwent either IVU or NCCT. Twenty patients with normal urinary tracts or with mild hydronephrosis proximal to the stone on urography comprised the IVU group. Eighteen patients who underwent NCCT and plain abdominal (KUB) films and had urinary tract systems similar in appearance to the IVU group comprised the NCCT group. The two groups were of similar mean age (45.75 years, range 24-73 years; and 49.0 years, range 26-72 years, respectively) and had a similar mean stone size (10.1-10.2 mm). Patients with internal ureteral or nephrostomy catheters were excluded. Information on episodes of intractable renal colic, urinary tract infections, and hospitalization was recorded at follow-up 2 to 6 weeks post-SWL. RESULTS: The IVU and NCCT patients had similar mean stone fragmentation rates (80% and 74%, respectively) at 2 to 6 weeks post-SWL. Four IVU patients (20%) had intractable renal colic. One NCCT patient (5.5%) had a urinary infection. Complication and hospitalization rates in the two groups were not significantly different (P = 0.34; Fisher' exact test). CONCLUSIONS: Using only NCCT before SWL was not associated with higher complication rates. Thus, IVU is not a prerequisite for SWL of radiopaque renal stones in patients with a normal urinary tract anatomy as seen on NCCT. PMID- 14744345 TI - Evaluation of chemical composition of urinary calculi by conventional radiography. AB - BACKGROUND AND PURPOSE: The choice of the most efficient treatment modality for renal calculi could be facilitated by determining the precise chemical stone composition before treatment. We investigated the possibility of using conventional X-ray imaging to determine stone composition and to find a simple method of predicting stone fragility for treatment planning. MATERIALS AND METHODS: The X-ray density of 92 stones with known infrared spectroscopy analyses (calcium oxalate monohydrate, calcium oxalate dihydrate, struvite, and calcium phosphate) was retrospectively investigated by scanning the films with a digital camera. The data analysis was done using a commercially available graphics program to compare the total gray-scale levels of the stones. RESULTS: There was a significant difference in the mean gray-scale level of calcium phosphate and calcium oxalate stones (P < 0.01). The mean gray-scale difference between calcium oxalate dihydrate and calcium oxalate monohydrate was also significant (p < 0.02). All calcium oxalate, struvite, and calcium phosphate stones were correctly identified. Of the calcium oxalate monohydrate and calcium oxalate dihydrate stones, 98.4% and 66.6%, respectively, were correctly identified. CONCLUSION: The method allows a reliable diagnosis of stone composition from radiographs, which can be useful in stone treatment planning. PMID- 14744346 TI - Correlation of ureteral stone measurements by CT and plain film radiography: utility of the KUB. AB - BACKGROUND AND PURPOSE: The practice of utilizing helical CT to evaluate patients suspected of renal colic is increasing. Little is known about the accuracy of CT in estimating stone size or the utility of an accompanying plain abdominal radiograph (KUB film). The purpose of our study was to compare ureteral stone size estimation by helical CT and plain film and determine whether a KUB film provides additional information useful in patient management. PATIENTS AND METHODS: Thirty consecutive patients (17 male, 13 female) having both a helical CT and a KUB study for evaluation of renal colic secondary to ureteral calculi comprised the study population. Calculus number, location, and dimensions were determined from these images. Stone dimensions were measured using electronic calipers on a picture archiving and communications system. Information found by KUB and CT was compared, and both sets of stone measurements were correlated with patient outcome. RESULTS: The mean maximal stone transverse diameter and length were similar on CT and plain film: 5.8 mm v 5.8 mm and 9.5 mm v 8.9 mm, respectively (P = 0.57 and 0.29, respectively). The mean anteroposterior stone diameter on CT of 6.8 mm was statistically greater than the transverse diameter as measured by both CT and KUB, which were 5.8 mm and 5.8 mm (P = 0.0002 and 0.0007, respectively). Eleven patients spontaneously passed their stones, while 19 patients required intervention. Patient outcome, as predicted by transverse stone width, was similar for CT and KUB data. CONCLUSIONS: The management of patients with ureteral calculi relies on estimated stone size and the stone's potential for spontaneous passage. Stone dimensions estimated by CT are similar to the size determined by plain film radiography. Although plain film radiography does not provide information on stone dimensions beyond that obtained with CT, it does reveal precise stone location and radiolucency, data helpful in following and treating patients. PMID- 14744347 TI - Limitations of noncontrast CT for measuring ureteral stones. AB - BACKGROUND AND PURPOSE: Patients with renal colic are frequently evaluated in the emergency room with a helical noncontrast CT scan (NCCT) as the primary imaging modality. Treatment decisions are often based on the size of the ureteral stone(s). We wished to assess the accuracy of NCCT in estimating ureteral stone size compared with plain abdominal (KUB) films. PATIENTS AND METHODS: Forty-eight patients were identified who had ureteral stones seen on NCCT and KUB films performed on the same day. The number of consecutive images on which a ureteral stone was visible on NCCT was multiplied by the reconstruction interval of 5 mm to create a size estimate, which was compared with the measurements of the same stone seen on the KUB film. RESULTS: The NCCT overestimated stone size by approximately 30% to 50% compared with KUB. CONCLUSION: Counting the number of consecutive NCCT images depicting a ureteral stone is not an accurate method of stone measurement when a reconstruction interval of 5 mm is used. Urologists should consider stone measurement techniques carefully and understand the limitations of imaging studies when evaluating patients with symptomatic ureteral stones. PMID- 14744349 TI - Philipp Bozzini--the father of endoscopy. PMID- 14744348 TI - Is there a role for pentosan polysulfate in the prevention of calcium oxalate stones? AB - The clinical role for pentosan polysulfate (PPS) in the prevention of calcium oxalate urolithiasis is not known. Crystallization and aggregation are important steps in calcium oxalate stone formation, and PPS has been shown to inhibit these steps, both in vitro and in vivo. In addition, PPS has a role in repairing injured urothelium and inhibiting adhesion to epithelial defects. A randomized double-blind placebo-controlled study appears warranted to assess the utility of PPS in the prevention of recurrent calcium oxalate stones. PMID- 14744350 TI - Comparison of clinical outcome of extracorporeal shockwave lithotripsy in patients with radiopaque v radiolucent ureteral calculi. AB - PURPOSE: To compare the efficacy of extracorporeal shockwave lithotripsy (SWL) in patients with radiopaque and radiolucent ureteral calculi. PATIENTS AND METHODS: Between September 2001 and August 2002, a total of 113 consecutive patients with ureteral calculi (58 radiopaque [Group 1], 55 radiolucent [Group 2]) were treated with a Multimed 2001 trade mark lithotripter under fluoroscopic monitoring. Localization of radiolucent stones was achieved after a bolus injection of contrast medium 1 mL/kg, the shockwaves being focused just below the end of contrast column. All patients were treated on an outpatient basis under analgesia and light sedation if required. The mean follow-up was 14 (range 6-23) months. Patients were reevaluated by some combination of plain films, ultrasound scanning, and intravenous urography 3 months after the treatment. Groups were compared with ANOVA and Mann-Whitney U tests. A P value <0.05 was regarded as statistically significant. RESULTS: There were no statistically significant differences in any parameter between groups. At 3 months, the success rate (stone free status) was 87.9% in Group 1 and 89% in Group 2 (P = 0.848). Double-J catheter replacement was needed for three patients in Group 1 and for two patients in Group 2 (P = 0.693). Two patients from Group 1 and three from Group 2 underwent intracorporeal lithotripsy with ureterorenoscopy (P = 0.606). No adverse reactions to contrast medium occurred in Group 2. CONCLUSION: Patients with radiolucent ureteral calculi can be treated efficiently with SWL by contrast medium injection if ultrasonic localization is not possible. PMID- 14744351 TI - Sticks and stones: use of acupuncture in extracorporeal shockwave lithotripsy. AB - BACKGROUND AND PURPOSE: Extracorporeal shockwave lithotripsy (SWL) is an effective noninvasive, outpatient method of stone clearance. In our unit, it is performed using a combination of oral analgesia and intravenous sedation, which allows us to treat to therapeutic levels in the vast majority of our patients. However, we have encountered patients who do not tolerate various elements of the analgesia protocol and thus cannot be treated to full effect. The options for these people are currently limited to either SWL under formal sedation or epidural or general anesthetic or the use of another technique of stone clearance, such as percutaneous nephrolithotomy, which may not be as appropriate, and again necessitates an anesthetic, an inpatient stay, or both. PATIENTS AND METHODS: We describe three patients who had previously failed SWL who received acupuncture in place of standard analgesia prior to the next treatment. RESULTS: All three patients were able to tolerate the procedure better and were treated at a higher level with more shocks than in the previous session. No side effects were noted. CONCLUSIONS: We propose that acupuncture may be considered in patients unable to take standard sedoanalgesia. It is a cost-effective, safe method of inducing sedation with analgesia and had no demonstrable side effects in our series. It provides an attractive alternative to the use of general or regional anesthetics in these patients. PMID- 14744352 TI - Routine stenting after ureteroscopy for distal ureteral calculi is unnecessary: results of a randomized controlled trial. AB - BACKGROUND AND PURPOSE: Insertion of a ureteral stent is routinely done after ureteroscopy. Recently, several authors have questioned routine stenting after ureteroscopy for distal ureteral stones. We report our results of a randomized study comparing ureteroscopy with and without placement of stents for distal ureteral stones. PATIENTS AND METHODS: A total of 48 patients undergoing ureteroscopy for distal ureteral stones were randomized to a stented group (N = 26) or a nonstented group (N = 22). Ureteroscopy was carried out with Wolf 8.5F semirigid endoscope, and the Swiss Lithoclast was used as the source of energy. Any stent was removed at 3 weeks. Patients were assessed for success, operative time, postoperative pain score, analgesic requirement, stent-related symptoms, and risk of ureteral stricture formation. Baseline variables were not significantly different in the two groups. RESULTS: There was no significant difference in the two treatment groups with regard to need for ureteral dilation, use of intracorporeal lithotripsy, or occurrence of intraoperative and postoperative complications. A successful outcome was achieved in 100% of both groups. The mean pain score on day 0 was 5.23 +/- 0.95 of 10 in the stented group and 4.82 +/- 0.96 in the nonstented group; this difference was not statistically significant. Similarly, the analgesic requirement in the two treatment groups was not significantly different. However, patients with stents had significantly more pain (including flank pain with voiding: P = 0.01), urgency (P = 0.04) and dysuria (P <0.01). Radiologic follow-up was available for 83.33% of the patients at the 3-month visit. None of the patients had evidence of ureteral stricture or residual stone fragments. CONCLUSION: In select patients undergoing ureteroscopy for distal ureteral stones, stents can be safely omitted. Patients without stents have significantly fewer lower-urinary symptoms of pain, urgency, and dysuria and are not at risk of increased complications. Avoiding stents may be particularly cost effective in developing countries. PMID- 14744353 TI - Nephrolithiasis associated with renal insufficiency: factors predicting outcome. AB - BACKGROUND AND PURPOSE: Renal calculous disease may be associated with various degrees of renal insufficiency secondary to a combination of obstruction, urinary infection, frequent surgical intervention, and coexisting medical disease. Herein, we present our data on the progression of renal function in patients with stones associated with renal insufficiency and assess the significance of various factors that could predict postoperative renal function deterioration. PATIENTS AND METHODS: Data were obtained from 4400 patients undergoing treatment for calculous disease at our institute since 1991. Renal insufficiency, defined as a baseline serum creatinine >1.5 mg/dL, was present in 84 (1.9%). Predictive factors evaluated for renal function deterioration were preoperative (age, duration of symptomatology and nephrolithiasis, urinary tract infection, coexistent medical diseases, baseline serum creatinine, and stone burden), intraoperative (number of percutaneous tracts), and postoperative (recurrent infection, proteinuria, cortical atrophy, residual fragments, and stone recurrence). RESULTS: Over a mean follow-up of 2.2 years (range 6 months-6 years), 33 patients (39.3%) showed improvement, 24 (28.6%) showed stabilization, and 27 (32.1%) showed deterioration in their renal function. Higher baseline serum creatinine, proteinuria >300 mg/day, renal cortical atrophy, stone burden >1500 mm(2), recurrent urinary infection, and age <15 years were significant predictors of subsequent renal function deterioration. CONCLUSIONS: Patients with nephrolithiasis and mild to moderate renal insufficiency warrant aggressive treatment aimed at complete stone clearance and prevention of recurrence of stones and urinary infection. A higher baseline preoperative serum creatinine, proteinuria >300 mg/day on follow-up, renal cortical atrophy, stone burden >1500 mm(2), recurrent urinary infection, and age <15 years are associated with a significantly higher likelihood of renal function deterioration after treatment of the calculous disease. PMID- 14744354 TI - Safety and efficacy of same-session bilateral ureteroscopy. AB - PURPOSE: Same-session ureteroscopy for bilateral urinary calculi would potentially reduce costs and the need for a second anesthetic compared with staged procedures. We sought to establish the safety and efficacy of same-session bilateral ureteroscopy relative to procedures for staged bilateral and multiple unilateral calculi in the context of contemporary instrumentation. PATIENTS AND METHODS: A series of 626 consecutive patients underwent ureteroscopy for calculi between January 1997 and August 2001. Among these, 34 patients with bilateral calculi (11 staged and 23 treated in one sitting) and 54 patients with multiple unilateral calculi in distinct locations were included in this study. Multivariable regression was used to determine the association of patient specific and technical factors with postoperative morbidity. RESULTS: Stone-free rates were similar in the two groups and ranged from 50% to 100% depending on stone location. Postoperative complications occurred in 6 (11%) and 3 (14%) of the patients treated for multiple unilateral and for bilateral calculi in a staged procedure, respectively, compared with 7 (29%) of those undergoing same session bilateral ureteroscopy (P = 0.12). Logistic regression revealed that same session bilateral ureteroscopy (odds ratio [OR] 4.0; P = 0.02) and absence of a postoperative stent (OR 1.7; P = 0.03) were associated with added morbidity. However, the cumulative risk of performing staged bilateral procedures (14% per procedure) approximated that of bilateral ureteroscopy in one sitting (29%). CONCLUSION: Bilateral ureteroscopy carries an increased risk of postoperative morbidity. The risk is proportional to the number of renal units treated and may be assumed at once (e.g., same-session) or over time (e.g., staged) as it applies to patients requiring bilateral ureteroscopy. PMID- 14744355 TI - Treatment of ureteral calculi with ballistic lithotripsy. AB - BACKGROUND AND PURPOSE: Ballistic lithotripsy is one of the new intracorporeal lithotripsy methods. In this study, the efficacy and complications of this method in the management of ureteral calculi were evaluated. PATIENTS AND METHODS: From November 1999 to December 2001, 340 patients (mean age 39.8 years; range 1.5-82 years) with a total of 362 ureteral calculi (bilateral in 22 cases) were treated with an 8.5F rigid ureteroscope and the Swiss Lithoclast. Of the calculi, 115 (32%) were located in the upper ureter, 63 (17%) in the middle ureter, and 184 (51%) in the lower ureter. The mean stone size was 10.4 mm (range 5-22 mm). RESULTS: Nearly all (344; 95%) of the calculi were accessible with the ureteroscope, and 321 calculi (88.7%) were fragmented completely, either with no residual fragments or with residual fragments <3 mm. In 3 cases (0.8%), there were residual fragments of about 4 mm after the procedure that passed spontaneously. Twenty calculi (5.5%) migrated to the kidney during the procedure and were subsequently treated with adjuvant SWL. Major complications occurred in 2 cases (0.54%): ureteral perforation and stenosis in 1 patient each. The 2-week stone-free rate was 89.5% (324/362). CONCLUSION: Lithoclast ballistic lithotripsy is a safe and effective approach for the treatment of ureteral calculi regardless of composition. PMID- 14744356 TI - Rapid communication: pure robot-assisted laparoscopic ureteral reimplantation for ureteral stricture disease: case report. AB - BACKGROUND AND PURPOSE: The role of the da Vinci robot is slowly being defined in minimally invasive urologic surgery. We report its use in the management of ureteral stricture disease. CASE REPORT: A 42-year-old man with recurrent kidney stone disease was found to have a left distal-ureteral stricture. After failure of endoscopic treatment, a robot-assisted laparoscopic ureteral reimplantation was performed. The total operative time was 210 minutes. The estimated blood loss was <50 mL. There were no intraoperative or postoperative complications. Total analgesic use was 30 mg of morphine. The hospital stay was 5 days. CONCLUSION: Pure robot-assisted laparoscopic ureteral reimplantation is a safe and feasible approach to the management of ureteral stricture disease. PMID- 14744357 TI - Prospective experience with a second-generation hand-assisted laparoscopic device and comparison with first-generation devices. AB - BACKGROUND AND PURPOSE: The GelPort Hand-Assisted Laparoscopy (HAL) device was licensed for use in the U.K. in September 2001. We compared our experience with this second-generation device with that of first-generation devices; i.e., the Handport, launched in 1999, and the Intromit, first marketed in 1998. MATERIALS AND METHODS: We prospectively compared a number of parameters for operations performed using the GelPort (13 radical nephrectomies, 4 nephroureterectomies) with those performed using the Handport (3 radical nephrectomies, 2 nephroureterectomies, 2 simple nephrectomies) and the Intromit (2 radical nephrectomies, 1 nephroureterectomy, 2 simple nephrectomies). The main outcome measures were ease of application, time required to place the device, and perioperative complications specific to the device. RESULTS: The device requiring the longest time to place was the Intromit (average 15 minutes) followed by the HandPort (average 10 minutes) and then the GelPort (average 5 minutes). There were two leaks with the Intromit (one major and one minor). Pop-outs were a frequent issue with the HandPort, necessitating repeated replacement and resufflation. There was also a need to resufflate every time the hand was removed for a change of swab. None of these problems was noted with the GelPort, which was also found to be the easiest to use. The major disadvantage of the GelPort was its price, which was about a third more than that of the first-generation devices. CONCLUSION: The GelPort is currently a more user-friendly and robust HAL device. It is, however, more expensive than first-generation devices. PMID- 14744358 TI - Octylcyanoacrylate skin closure in laparoscopy. AB - BACKGROUND AND PURPOSE: Octylcyanoacrylate (Dermabond) is a tissue "glue" useful in closing surgical skin incisions. We compared skin octylcyanoacrylate with subcuticular skin sutures to close laparoscopic trocar sites. PATIENTS AND METHODS: A randomized double-armed study was performed with 59 patients in whom 228 trocar sites were closed. Twenty-nine patients underwent subcuticular closure with 4-0 absorbable suture, and thirty patients received closure with octylcyanoacrylate in accordance with the recommendations of the manufacturer (Ethicon; Somerville, NJ). The number of sutures or vials of octylcyanoacrylate used, closure times, and postoperative wound problems were recorded. Wounds were assessed 2 weeks postoperatively for healing complications. Closure costs were estimated using published operating room time per hour plus the cost of octylcyanoacrylate or suture. Student's paired t-test was used for statistical analysis. RESULTS: The overall mean time required for skin closure using octylcyanoacrylate and suture was 3.7 minutes and 14 minutes, respectively (P <0.00001). An average of 2.2 packets of suture were used to close all port sites in a particular patient, while closure with octylcyanoacrylate required an average of 3.4 vials per patient. Wound complications consisted of subcuticular seroma with skin separation and were equally common in the two groups. The overall average cost per closure using octylcyanoacrylate was $198 US dollars, while the cost for closure using suture was $497 US dollars (P <0.00001). CONCLUSIONS: Laparoscopic port-site skin closure with octylcyanoacrylate is rapid and effective. The method yields cost savings and a decrease in operative time of more than 9 minutes per case. PMID- 14744359 TI - Laparoscopic partial nephrectomy in a horseshoe kidney. AB - A 68-year-old man with an incidentally found 2-cm complex enhancing cystic right renal mass in the right moiety of a horseshoe kidney was treated with a three port retroperitoneal laparoscopic approach. The tumor was completely excised with cold Endoshears, and Surgicel bolsters were tightly buttressed into the resection bed with 0 Vicryl sutures. The warm ischemia time was 31 minutes. To our knowledge, this is the initial case of retroperitoneal laparoscopic partial nephrectomy in a horseshoe kidney. Three-dimensional CT with volume rendering in a video format provides the necessary information about the number, location, and extrarenal anatomy of the renal artery and vein. PMID- 14744360 TI - Retroperitoneoscopic nephrectomy of a horseshoe kidney with renal-cell carcinoma. AB - A 55-year-old man had a solid tumor in the right part of a horseshoe kidney. He had previously undergone open cholecystectomy and appendectomy with panperitonitis. Retroperitoneoscopic nephrectomy was performed, with rapid recovery and good short-term results. This appears to be the first case of endoscopic surgery on a horseshoe kidney using ultrasonic shears for cancer. PMID- 14744361 TI - Hand-assisted laparoscopic pyeloplasty and isthmectomy in a patient with a horseshoe kidney. AB - We report the laparoscopic management of ureteropelvic junction obstruction in a patient with a horseshoe kidney. We believe this to be the first reported case of isthmectomy performed with the Harmonic Scalpel. Follow-up diuretic renal scan and CT revealed improved drainage and separation of the divided horseshoe kidney. PMID- 14744362 TI - Laparoscopic radical cystectomy and ileal conduit reconstruction: preliminary experience. AB - PURPOSE: We present our preliminary experience with laparoscopic radical cystoprostatectomy for muscle-invasive carcinoma of the urinary bladder. Patient and operative data and the surgical technique are presented. PATIENTS AND METHODS: Laparoscopic radical cystoprostatectomy and bilateral pelvic lymph node dissection were performed using five or six ports by a transperitoneal approach. An ileal conduit urinary diversion was constructed at the site of specimen retrieval. RESULTS: The procedure was successful in nine of ten patients with a mean blood loss of 533 mL and an average transfusion of 1.3 units per patient. The mean operating time was 6.48 hours and an average of 33 mg of morphine equivalents was required for analgesia. The mean hospital stay was 10.8 days. One patient had surgical margins positive for cancer, while none had histologic evidence of pelvic nodal metastasis. There were five minor and major intraoperative and postoperative complications. The remaining patient, treated early in our experience, developed hypercarbia necessitating conversion to open surgery. No metastases have been seen after a mean duration of follow-up of 19 months. CONCLUSIONS: Laparoscopic radical cystoprostatectomy with open ileal conduit urinary diversion is a feasible alternative to traditional open radical cystectomy. Urinary diversion can be performed through the small incision necessary to extract the surgical (radical cystoprostatectomy) specimen from the abdomen. With our modified technique, it also is feasible to reduce the cost. PMID- 14744363 TI - Endoscopic management of milk of calcium-filled ureterocele stump. AB - A 38-year-old woman with a duplicated right collecting system and a history of right upper-pole heminephrectomy was referred for persistent dysuria and right lower-quadrant abdominal discomfort. Imaging identified a remnant ureter and a ureterocele filled with what appeared to be a large homogenous stone. At cystoscopy, the ureterocele was incised with a holmium:YAG laser, releasing a large quantity of white milky fluid (milk of calcium). There was no evidence of any solid material. Endoscopic evaluation should be the first step in patients with stones in a ureteral stump because milk of calcium may be the etiology of what appears to be a large stone burden in an obstructed system. PMID- 14744364 TI - Ureteral stricture developing after partial nephrectomy with a microwave tissue coagulator: case report. AB - A 32-year-old man with a 1.7-cm tumor in the left kidney underwent laparoscopy assisted partial nephrectomy. Although his postoperative course was uneventful, a 6-month postoperative CT scan showed hydronephrosis secondary to a severe stricture at the ureteropelvic junction. Heat injury to the urinary tract was strongly suspected. The use of microwave tissue coagulator for the tumor close to the renal sinus and excessive coagulation should be avoided to prevent heat related complications. PMID- 14744365 TI - Subsurface tissue lesions created using an Nd:YAG laser and cryogen cooling. AB - BACKGROUND AND PURPOSE: Cooling methods are commonly used in dermatology to protect the skin surface during cosmetic laser procedures. This study investigated deep laser heating of tissue in combination with cryogen cooling of the surface for potential noninvasive targeting of subsurface tissue structures in urology. MATERIALS AND METHODS: Porcine liver and skin were studied because they represent simple and difficult tissue models, respectively, for subsurface lesion production. Gross and histologic methods were used to measure lesion dimensions. RESULTS: Liver lesions measured 4.9 +/- 1.2 mm in diameter, while preserving the tissue surface to a depth of 2.1 +/- 0.4 mm (N = 5). Surface temperatures remained below 40 degrees C, while lesion temperatures ranged from 60 to 70 degrees C. Skin lesions measured 4.1 +/- 0.6 mm, while preserving the skin surface to a depth of 1.2 +/- 0.2 mm (N = 4). CONCLUSION: This technique may prove useful to target subsurface anatomic structures in urology. PMID- 14744366 TI - Bipolar radiofrequency ablation of the kidney: comparison with monopolar radiofrequency ablation. AB - PURPOSE: We report initial ex vivo and in vivo studies using bipolar radiofrequency (RF) ablation of porcine kidneys. An internal ground electrode is positioned in the kidney opposite the RF electrode, resulting in ablation of all the intervening renal tissue. MATERIALS AND METHODS: Ex vivo preparations of 10 porcine kidneys were perfused continuously with Ringer's solution and treated with either standard external grounded RF (N = 3) or bipolar RF ablation with 1 (N = 2), 2 (N = 3), or 3 (N = 2) cm of separation between the ground probe and the RF probe using a Model 30 RITA generator (RITA, Mountain View, CA). Target temperatures were 90 degrees C for 8 minutes. Gross and histologic assessments were made acutely. Four domestic pigs were treated with monopolar RF ablation of the lower pole of one kidney and bipolar RF with a 12-mm separation between the probes of the contralateral lower pole. Animals were harvested 48 hours later to maximize tissue damage for gross measurements and histologic evaluation. RESULTS: Ex vivo studies revealed grossly monopolar lesions 1.5 cm in maximum diameter and 1.75 cm(3) in volume. In comparison, bipolar lesions were 2.8 cm in maximum diameter and 10.3 cm(3) in volume using 3 cm of electrode separation. There was histologic evidence of cell death in all specimens. In vivo studies showed two distinct gross lesions with RF: one blanched and one hemorrhagic. Using bipolar RF, larger blanched lesions were achievable than with monopolar RF (2.80 cm(3) v 1.63 cm(3)). Overall, the combinations of blanched and hemorrhagic lesions were similar with monopolar and bipolar RF (5.01 v 5.31 cm(3)). Histologic evaluation verified cell death in the blanched lesions and rare areas of normal tissue in the hemorrhagic lesions. CONCLUSIONS: As shown by ex vivo data, bipolar RF can create larger lesions than does monopolar RF. In vivo, at 48 hours, both blanched and hemorrhagic gross lesions were seen using RF. In this model, blanched lesions predominated when performing bipolar RF. PMID- 14744367 TI - Effect of retroperitoneoscopic donor nephrectomy on tissue oxidative stress markers in rabbit pneumoretroperitoneum model. AB - BACKGROUND AND PURPOSE: Laparoscopic donor nephrectomy causes increased renal oxidative stress. There are no data about the effects of the retroperitoneoscopic route. The aim of our study was to evaluate the oxidative stress occurring in renal tissues during retroperitoneoscopic donor nephrectomy in a rabbit model. MATERIALS AND METHODS: Eighteen adult rabbits were randomized to three groups, each consisting of six rabbits. Group I (control) underwent 50-mL balloon dissection of the left retroperitoneal space without CO(2) insufflation. Group II (pneumoretroperitoneum) received a 3-hour CO(2) insufflation at a pressure of 10 mm Hg in the retroperitoneal space after balloon dissection. Group III (pneumoretroperitoneum with warm ischemia), in addition to the procedure applied in Group II, underwent left renal artery clamping for 3 minutes and reperfusion for the next 5 minutes. Bilateral nephrectomy was performed in all animals for analysis of oxidative stress markers. Concentrations of malonyldialdehyde (MDA), protein carbonyl, and reduced glutathione (GSH) were measured in renal tissue samples. RESULTS: The MDA and protein carbonyl content were increased both in the donor (P = 0.004 and P = 0.004, respectively) and in the remaining kidneys (P = 0.009 and P = 0.028, respectively) in Group II compared with Group I. There were no statistically significant increases in oxidative stress markers between Group II and Group III in donor kidneys. However, there were statistically significant decreases in MDA in the remaining kidneys in Group III compared with Group II (P = 0.009). CONCLUSION: Pneumoretroperitoneum causes increased oxidative stress in both donor and remaining kidneys. Short-term warm ischemia and reperfusion do not exert an additive effect on pneumoretroperitoneum-associated oxidative stress in donor kidneys. PMID- 14744368 TI - Retrograde vaginal methylene blue injection for localization of complex urinary fistulas. AB - BACKGROUND: Ureterovesical and vesicovaginal fistulae usually are easily diagnosed with standard techniques, including radiology, double-dye test, cystoscopy, and vaginal examination. In a few complex cases, however, the localization of the fistulous orifice may be extremely difficult. METHOD: We describe a novel technique of retrograde vaginal methylene blue testing that may assist in the proper localization of the fistulous opening. RESULTS: Our experience with one case each of ureterovesical and vesicovaginal fistula easily diagnosed with this method proves its efficacy, simplicity, and safety. CONCLUSION: This technique should be reserved for cases of urinary fistulae when all standard diagnostic methods have failed. PMID- 14744369 TI - Percutaneous peritoneal drainage for intraperitoneal bladder perforations during transurethral resection of bladder tumors. AB - PURPOSE: To demonstrate the safety and efficacy of percutaneous drainage of the peritoneal cavity using an 8F pigtail catheter placed under ultrasound guidance in cases of intraperitoneal perforation of the bladder as a result of transurethral resection of a bladder tumor (TURBT). PATIENTS AND METHODS: Three patients undergoing TURBT had inadvertent but significant intraperitoneal perforations of the bladder. All patients developed signs and symptoms of peritonism despite large catheters draining the bladder. An 8F pigtail catheter (Boston Scientific, Watertown, MD, USA) was inserted percutaneously under ultrasound guidance with local anesthetic infiltration. RESULTS: All three patients made a good recovery without needing a laparotomy. Cystograms a week later confirmed an intact bladder. Clinical review revealed no sequelae as a result of the perforation or its treatment. CONCLUSION: Percutaneous drainage of the abdomen in patients with intraperitoneal perforation of the bladder during TURBT is a safe alternative to laparotomy. PMID- 14744371 TI - [Enhance the application of pediatric critical illness score]. PMID- 14744372 TI - [Pediatrician should regard to the prevention and treatment of HIV infection and acquired immunodeficiency syndrome]. PMID- 14744373 TI - [Clinical application of simplified pediatric critical illness scoring system]. AB - OBJECTIVE: To simplify the Pediatric Critical Illness Scoring (PCIS), to evaluate the simplified PCIS and to make the simplified scoring system applicable in the grassroots hospitals. METHODS: Totally 1,036 patients were scored by PCIS on 1, 3, 7 and the last (discharged or dead) hospital days. The PCIS has 10 items. The full score is 100. The scores of approximately 100, approximately 80, and approximately 70 represent non-serious, serious, and extremely serious patients' condition. The PaO(2) and pH (2 items), BUN or creatinine (3 items), plasma sodium and potassium (5 items) were deleted from PCIS's 10 items in turn. The remaining 8, 7, and 5 items were applied as new scoring system after the original scores were multiplied by 0.8, 0.7, and 0.5. The simplified PCIS was evaluated by comparing the patients' condition that was assessed by PCIS. The consistency rate of patients' condition that was obtained by using PCIS and simplified PCIS should be over 80%. The mortality of non-serious, serious, and extremely serious groups that were defined by using PCIS and simplified PCIS was observed and compared. RESULTS: When 2, 3 and 5 items were deleted from the PCIS on d1 scoring the consistency rates of the patients' condition were 82.6%, 80.7%, and 69.9%. While 5 items remained on d1 scoring the consistency rate was lower than 80%. When the same 2, 3 and 5 items were deleted from PCIS on d3, d7 and last scoring, the consistency rates of the patients' condition were 81.5% approximately 97.1%. The PCIS and simplified PCIS had a close correlation (r = 0.629-0.948, P < 0.001). In PCIS the mortality rates of non-serious, serious and extremely serious patients were significantly different. When simplified PCIS was used, the mortality rates of the three groups also had significant differences (chi(2) = 86.13-740.33, P < 0.001). Within a group of patients with the same condition, the mortality rates were not significantly different for PCIS and simplified PCIS. For instance, on d1 and last scoring, the mortality rates of extremely serious patients were 29.8%, 67.3% for PCIS and 30.0%-27.9%, 66.3%-64.4% for simplified PCIS. CONCLUSION: When 2 items (PaO(2) and pH) were deleted from d1 PCIS scoring and 5 items (PaO(2) and pH, BUN or creatinine, plasma sodium and potassium) were deleted from d3, d7, and last PCIS scoring, the results of assessment of patients' condition were basically the same as those of PCIS. The consistency rates of PCIS and simplified PCIS were > 80%. When simplified PCIS was applied, mortality rates of non-serious, serious, and extremely serious patients were significantly different that were the same as those of PCIS. In patients with the same condition, the mortality rates were not significantly different between the simplified PCIS and PCIS. PMID- 14744374 TI - [Effects of inhaled nitric oxide on rabbits with meconium aspiration pneumonia]. AB - OBJECTIVE: To evaluate effects of inhaled nitric oxide (iNO) on the expression of lung neutrophil adhesion molecule CD(11b) in experimental meconium aspiration pneumonia treated with conventional mechanical ventilation under room air or 100% O(2). METHODS: Rabbits were randomly allocated to 10 groups (n = 60), 6 of each group. Control or meconium aspiration pneumonia model groups were inhaled with room air or 100% O(2). Six treatment groups were treated with continuous NO inhalation at the doses of 6 x 10(-6), 10 x 10(-6) and 20 x 10(-6), respectively for 12 hours under room air or 100% O(2). The ratio of wet/dry (W/D) lung weight, alveolar septal width (ASW), myeloperoxidase (MPO) activity and lung injury score were measured. The expression of CD(11b) in neutrophils of the bronchoalveolar lavage fluid (BALF) was detected with flow cytometry. RESULTS: After 12 hours ventilation, the oxygenation was maintained better in treatment groups under different O(2) concentrations than that in model groups. Inflammatory evidence was found in lungs from all the model groups and treatment groups, which was characterized by serious inflammatory cell infiltration in alveolar space and hyaline membrane formation. The lung inflammation was decreased in all groups with nitric oxide inhalation. The ratio of W/D lung weight and ASW among different groups had no significant difference. MPO activities were significantly decreased in groups treated with 10 x 10(-6) and 20 x 10(-6) iNO compared with the model groups [with the concentration of 21% O(2), (1.8 +/- 0.2) U/g vs (4.4 +/- 0.5) U/g and (2.0 +/- 0.1) U/g vs (4.4 +/- 0.5) U/g;with the concentration of 100% O(2), (1.7 +/- 0.4) U/g vs (2.8 +/- 0.5) U/g and (1.4 +/- 0.3) U/g vs (2.8 +/- 0.5) U/g, P < 0.05, respectively]. MPO activities in the 20 x 10(-6) iNO group under 100% O(2) were significantly reduced compared with those under 21%O(2) [(1.4 +/- 0.3) U/g vs (2.0 +/- 0.1) U/g, P < 0.05]. Nitric oxide inhalation with the doses of 10 x 10(-6) and 20 x 10(-6) significantly decreased the expression of CD(11b) (MFI) in neutrophils of the BALF compared with the expressions in model groups without NO treatment (with 21% O(2), 121 +/- 20 vs 392 +/- 204 and 112 +/- 30 vs 392 +/- 204; with 100% O(2), 113 +/- 24 vs 293 +/- 65 and 102 +/- 14 vs 293 +/- 65, P < 0.05, respectively). Under the same iNO dose (10 x 10(-6) or 20 x 10(-6)) no statistic difference was found between groups of different inspired oxygen concentrations (21% and 100%). CONCLUSIONS: Inhaled nitric oxide with the doses of 10 x 10(-6) to 20 x 10(-6) could significantly down-regulate the CD(11b) expression in neutrophil of the BALF and reduce the neutrophil sequestration and MPO activity in rabbit lungs, which may decrease the lung inflammation process in meconium aspiration pneumonia. PMID- 14744375 TI - [Clinical analysis of pediatric SARS cases in Beijing]. AB - OBJECTIVE: To study clinical characteristics of pediatric SARS cases in Beijing. METHODS: Eighteen pediatric cases with SARS diagnosed on admission were analyzed. The cases were admitted to Beijing Children's Hospital and Ditan Hospital (pediatric ward) from April 8 to May 12. RESULTS: The 18 children aged 5 months to 15 years (10 male and 8 female) had epidemiologically linked findings. Fourteen cases had close contact with SARS patients. Four cases were living in the community where adult SARS patients were found. All the 18 patients but one presented with fever and cough. Most of them had high fever, 2 cases had bradycardia, 2 had diarrhea, and another 2 had tachypnea. Malaise and headache were noted only in 3 cases respectively which were much less frequently seen than in adult patients. Symptoms and signs of the children were much less severe and aggressive than adults cases. Thirteen children had chest radiographic consolidation. Of them, 9 cases had progressive changes after admission, then improved quickly. We did not find significant lower hemoglobin and platelet levels. Most patients had leukopenia and lymphopenia. Serologic test was performed for 15 cases and 8 were positive for SARS virus-IgG and 6 for IgM antibody. Of the 4 cases who had close contact with SARS adults and without chest radiograph abnormal findings, 3 were negative for SARS virus-antibodies. Part of the patients had temporary abnormality of myocardial enzyme and liver function. All these children finally had rapid improvement on chest radiograph. The patients were treated with antiviral agents and corticosteroid. Only two cases required oxygen therapy. No child needed assisted ventilation and no death, nor lung fibrosis occurred. After hospitalization, all patients were improved and discharged when this paper was being written. The average hospital stay of these patients was 14.6 days (6 - 22 days). CONCLUSION: Compared with adults, pediatric SARS patients seemed to have their own clinical characteristics. The disease in children had lower severity and infectivity than that in adults. The mechanisms of the disease in children should be studied in well-designed clinical trials. Cases like the 4 children who had close contact with SARS adult patients but without chest radiographic changes deserve further studies with the help of more reliable and sensitive etiologic tests. PMID- 14744376 TI - [Development of a gap ligase chain reaction for detection of Chlamydia trachomatis in newborn infants]. AB - OBJECTIVE: To establish a gap ligase chain reaction (G-LCR) assay for the detection of Chlamydia trachomatis (Ct) in neonates with pneumonia. METHODS: A G LCR DNA amplification assay that targeted the outer major membrane protein gene (omp1) of Ct was developed to detect Ct. The sensitivity and specificity of the G LCR test was examined by the use of highly purified elementary bodies (EBs). Nasopharyngeal swabs taken from 328 neonates with pneumonia were analyzed by Gap LCR and cell culture. RESULTS: The detection limit of G-LCR was 2 EBs. G-LCR could detect five species of Ct and was not cross-reacted with C psittaci and other bacteria. The prevalence of Ct in 328 neonates with pneumonia, using an expanded gold standard of a positive cell culture or two confirmed positive non culture tests, was 21% (69/328). After analysis of discrepant results, the sensitivity, specificity, and positive and negative predictive values for the G LCR were 98.6%, 100%, 100% and 99.6%, respectively; whereas those for culture were 86.9%, 100%, 100% and 96.6%, respectively. CONCLUSION: This study demonstrated that the G-LCR was a highly sensitive nonculture technique and good alternative test for the detection of chlamydial infections. PMID- 14744377 TI - [Activation of nuclear factor kappa B in newborn rats sepsis]. AB - OBJECTIVE: The aim of the study is to explore the effect of NF-kappa B signal pathway in neonatal sepsis so as to provide the experimental base for corresponding clinical treatment of the sepsis, in which NF-kappa B is taken as the target. METHODS: The sepsis model was established in newborn rats by giving Staphylococcus aureus subcutaneously: (1) The electrophoretic mobility shift assay (EMSA) was used to observe the activity of NF-kappa B in the lungs and the livers in newborn rats with Staphylococcus aureus sepsis. (2) Immunohistochemical method was used to observe the activity of NF-kappa B P56 in newborn rats with Staphylococcus aureus sepsis. (3) The anti-oxidant pyrrolidine dithiocarbamate (PDTC) was used to observe its effect on NF-kappa B activities of liver and lungs and on the activity of splenic NF-kappa B P56 in newborn rats with Staphylococcus aureus sepsis. RESULTS: In newborn rats with Staphylococcus aureus sepsis, the NF kappa B activity in lungs was enhanced at the 1st hour and reached to the peak level at the 3rd hour; then, it was weakened gradually and at the 24th hour faded away. The activity of the liver NF-kappa B was also activated and peaked at the 4th hour; then, it was gradually weakened and at the 24th hour faded away. The positive expression of splenic NF-kappa B P56 began to be intensified at the 1st hour (12.0 +/- 3.7), peaked at the 3rd hour (51.4 +/- 5.9) and showed insignificant differences at the 24th hour (3.4 +/- 1.4) as compared with the sepsis group. PDTC had an inhibitive effect on the activities of liver NF-kappa B and lung NF-kappa B and on the positive expression of splenic NF-kappa B P56 used in the dosage of 50-200 mg/kg. The larger the dosage was used, the more intensified inhibitive effect could be obtained. In the dosage of 200 mg/kg, the inhibitive effect was the most intensified. CONCLUSIONS: (1) In newborn rats with Staphylococcus aureus sepsis, the NF-kappa B of lungs, liver and spleen were activated, and all indicate a peak. (2) The anti-oxidant PDTC can inhibit NF kappa B activity in a dose-effect fashion in newborn rats with Staphylococcus aureus sepsis. PMID- 14744378 TI - [A survey of children with HIV/AIDS in highly epidemic villages of AIDS]. AB - OBJECTIVE: To estimate prevalence of HIV/AIDS among children and the transmission routes in a highly endemic villages of AIDS. METHODS: Totally 208 high-risk women of child bearing age and 159 of their children aged 0 - 14 years were investigated. Their medical histories of blood donation or transfusion were collected, blood samples were taken and sera were separated for HIV test. Enzyme linked immunosorbent assay (ELISA) and Western blot assay were performed for HIV antibody. The Nested-polymerase chain reaction (PCR) assay amplifying gag gene p17 was performed on samples of children aged less than 18 months. RESULTS: Thirty-seven HIV infected cases were found among 159 children aged 0 - 14 years of whom 33 were infected by mother-to-child transmission (89.2%, 33/37), 3 by blood transfusion (8.1%, 3/37) and one by iatrogenic route (2.7%, 1/37). Sixty seven mothers who were seropositive for HIV and their 86 children who were born after 1992 were investigated, 33 cases of them were infected with HIV. The rate of vertical transmission was 38.4% (33/86). The HIV vertical transmission rate among mothers with AIDS (68.8%, 22/32) was significantly greater than that among mothers with asymptomatic HIV infection (20.4%, 11/54, P < 0.05). The number of children infected with HIV through vertical transmission increased from 1993 to 2001. Among 37 children infected with HIV, 12 cases developed AIDS and 4 of them died, of whom 2 cases died from tuberculosis. The morbidity of AIDS was 27.3% (9/33). Ninety three point nine percent (31/33) of infected mothers didn't know their HIV seropositive status before pregnancy and delivery. Of 8 pregnant women infected with HIV, one had aggravation of AIDS, 2 miscarried, 2 terminated their pregnancy and 3 continued their pregnancy. CONCLUSION: Mother-to-child transmission of HIV was the major route of HIV/AIDS transmission to the children. The main reason leading to HIV infection in children was the lack of prenatal HIV counseling and testing for the high-risk women of childbearing age and lake of interventions. The countermeasures must be taken to control the further transmission of AIDS in order to protect the health of women and children in the highly endemic areas of AIDS. PMID- 14744379 TI - [A case of MN hemolytic disease in newborn]. PMID- 14744381 TI - [A case of hepato-glycogenosis of newborn]. PMID- 14744380 TI - [Identification of two novel WASP gene mutations in 3 boys with Wiskott-Aldrich syndrome]. AB - OBJECTIVE: The Wiskott-Aldrich syndrome (WAS) is a rare X-linked primary immunodeficiency caused by mutations in the WAS protein (WASP) gene. The disease is characterized by recurrent infections, eczema, and thrombocytopenia with small platelets, and it is known to be associated with extensive clinical variability, and mutation studies indicated that genotypes are also highly variant among WAS patients. The present study was conducted to identify the mutation types of Wiskott-Aldrich syndrome protein (WASP) gene in 3 boys suffering from Wiskott Aldrich syndrome. METHODS: Based on the typical clinical manifestations of Wiskott-Aldrich syndrome including thrombocytopenia, eczema, and recurrent infections and scanning electron micrographs, 3 patients were suspected of having WAS. The WASP gene of the 3 patients and their mothers were detected by PCR direct sequencing analysis. RESULTS: By sequence analysis using sense and antisense primer separately, the authors found two novel WASP gene mutations. For the twin brothers, a C deletion at nucleotide 984 was detected in exon 10 of WASP gene (984delC). The consequence of the C deletion involved frameshift mutation after H317 and premature stop at 444 (H317fsX444). Their mother was a carrier of the mutated WASP gene. For another WAS patient, a nonsense mutation with nucleotide substitution of G to T at position 1388 (1388G-->T) in exon 11 of WASP gene, led to premature translational termination at amino acid position 452 (E452X). His mother had not been found to have WASP gene mutation. CONCLUSION: Genetic analysis is useful in definite diagnosis of Wiskott-Aldrich syndrome patients and in carrier detection and prenatal diagnosis, especially of atypical or sporadic WAS patients. PMID- 14744382 TI - [Inhibition of respiratory syncytial virus replication in cultured cells by RNA cleaving DNAzyme]. AB - OBJECTIVE: DNAzyme/Deoxyribozyme is another novel molecular biological tool following the ribozyme. DNAzyme consists of a 15-nucleotide (nt) internal loop as its catalytic domain and two flanking substrate-recognition domains of 7 to 8 nt each which is complementary to substrate. The RNA substrate is cleaved at a particular phosphodiester located between an unpaired purine and a paired pyrimidine residue. DNAzyme has been applied in fields such as viral infectious disease, tumor, cardiovascular disease and genetic disease. But there is no report about using DNAzyme for anti-respiratory syncytial virus purpose. To observe the inhibitory effects of RNA-cleaving DNAzymes on respiratory syncytial virus (RSV) replication in cultured cells. METHODS: Anti-RSV RNA-cleaving DNAzyme DZ604 was designed to target the RSV genome at the start of the NS2 gene in an effort to inhibit the RNA replication. Microscope and electron microscope were used to observe the effects of anti-RSV genomic RNA DNAzyme on cytopathogenic effect (CPE) and ultrastructural change of 9HTE cell induced by RSV infection. Viral plaque forming reduction assay and MTT assay were used to detect the anti RSV activity and protective function for RSV infected 9HTE cells of DNAzyme. RESULTS: Anti-RSV genomic RNA DNAzyme (DZ604) significantly improved CPE of RSV infected 9HTE cells. The time to appearance of CPE and of total CPE was delayed by using DZ604 in a dose-dependent manner. At a 5 micro mol/L concentration of DZ604, CPE of 9HTE cells induced by RSV infection at 10 and 1 multiplicity of infection (MOI) was not improved. At smaller MOI (0.1, 0.01, 0.001, 0.0001) of RSV infection, CPE of 9HTE cells was significantly lightened by DZ604 at the same concentration. DZ604 also significantly improved ultrastructural change of 9HTE cells at early stage of RSV infection. Reduction in RSV yield was 85.56% and 8.33% at concentrations of 5 micro mol/L and 0.25 micro mol/L of DZ604. DZ604 inhibited RSV yield in a dose-dependent manner (P < 0.05). Non-specific DNAzyme did not have anti-RSV activity (P > 0.05). CONCLUSION: Anti-RSV genomic RNA DNAzyme designed and synthesized in our laboratory was capable of inhibiting respiratory syncytial virus replication specifically in cultured cells. Our data indicated that DNAzymes could be useful for the prevention against respiratory syncytial virus infection. PMID- 14744383 TI - [The correlation between Kawasaki disease and polymorphisms of Tumor necrosis factor alpha and interleukin-10 gene promoter]. AB - OBJECTIVE: Kawasaki disease (KD) is an acute febrile vasculitic syndrome of unknown etiology that preferentially affects coronary artery. It has been suggested that proinflammatory cytokines like tumor necrosis factor alpha (TNF alpha) and interleukin-10 (IL-10) are key players during acute KD. Recently, the polymorphisms relative to major transcriptional start site of TNF-alpha and IL-10 gene were shown to influence the level of TNF-alpha and IL-10 production in vitro. This study was aimed to investigate the genetic association of TNF-alpha and IL-10 promoter polymorphisms in juvenile patients of Han nationality with KD, and to investigate the possible associations with clinical manifestations of the disease. METHODS: Four polymorphism sites of TNF-alpha and IL-10 gene promoter regions from 96 children with KD were determined by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP). One hundred and sixty age matched normal children of the Han nationality were used as control. All patients accepted Doppler echocardiography examination in order to differentiate coronary artery lesions. RESULTS: There was significant difference in allele frequencies of -308 (A/G) site of the TNF-alpha gene between children of the Han nationality and those of Japanese and Caucasian in America. There were significant differences in the allele frequencies of -1082 (G/A), -819 (C/T) and -592 (A/C) of IL-10 gene between children of the Han nationality and their British Counterparts (P < 0.01). There was no significant difference in allele frequencies of -308 (A/G) site of TNF-alpha gene between children with KD and normal controls. There was no significant difference in the haplotypes and the allele frequencies of the above three sites of IL-10 between the two groups. However, when clinical features were examined, the genotype frequency of TNF alpha-308A was significantly higher in IVIG-resistant KD patients than that of TNF-alpha-308G genotype (67% vs 5%, chi(c)(2) = 90.48, P < 0.01). The genotype of TNF-alpha-308A was closely associated with IVIG-resistant KD (P < 0.01, relative risk 42.25, 95% confidence interval 15.81-112.88). The haplotype frequency of IL 10 -1082A/-819T/-592A was also higher in patients with coronary artery lesion (CAL) caused by KD than those of Non-ATA haplotype (52% vs 20%, chi(2) = 18.36, P < 0.01). The haplotypes of IL-10 -1082A/-819T/-592A was significantly associated with CAL caused by KD (P < 0.01, relative risk 4.26, 95% confidence interval 2.20 8.25). CONCLUSION: The genotype of TNF-alpha-308A is one of the important factors that probably influence the therapeutic effect of KD. The haplotypes (-1082/-819/ 592) of IL-10 gene promoter might be related to the pathogenesis of coronary artery complication of KD and -1082A/-819T/-592A haplotypes might be regarded as a genetic marker of risk factor for coronary artery lesion in KD. PMID- 14744385 TI - [The expansion and biological characteristics of human mesenchymal stem cells]. AB - OBJECTIVE: Mesenchymal stem cells (MSCs) were adult stem cells which contribute to the regeneration of mesenchymal tissues such as bone cartilage, muscle, ligament, tendon, adipose and stroma. Due to the multipotential ability and self renewal capacity, the mesenchymal stem cells can be applied in many fields, such as the seed cells in tissues engineering, cell therapy and gene therapy. To enhance the clinical use of MSCs, the investigators studied the isolation and expansion of MSCs from adult bone marrow, fetal bone marrow and human umbilical cord blood, and investigated their biological identities. METHODS: Two kinds of incubation systems containing L-DMEM or MSC special culture medium were used to purify and expand MSCs. The growth, purification and proliferative abilities of 3 kinds of MSCs were observed and their immunophenotypes were determined by flow cytometry. RESULTS: (1) The shapes of 3 kinds of cells were same. There was no difference in number and size. The colonies formed early in adult bone marrow MSCs. (2) There was no difference in the expansion speed of the 3 kinds of MSCs, but after the colonies confluenced there had no touching constrain in MSCs from umbilical cord blood and fetal bone marrow. When the colonies confluenced, the cells also had proliferation ability. But in adult bone marrow, the touching constrain was significant. (3) MSCs had strong self-renewal capacity. After primary culture approximately 5 - 6 x 10(5) MSCs were obtained from 8 x 10(6) MNC of bone marrow and 25 x 10(6) MNC of umbilical cord blood. After passage 3, passage 5 and passage 10, the investigators could get 10(7), 10(8) and 10(10) MSCs, respectively. (4) Along with the increase in the passage and prolonging of culture time, the ability of expansion decreased, but they maintained good puripotentiality. After passage 2, passage 3 and passage 5, the purity of MSCs was 90%, 95% and 99%, respectively. (5) Three kinds of MSCs were all positive for CD(29), CD(44), CD(59), CD(90), CD(105), CD(166) and all negative for the markers of hematopoietic cells such as CD(11a), CD(14), CD(33), CD(34), CD(28), CD(45). All the important GVHD correlation markers were negative, such as HLA-DR, B7-1 (CD(80)), B7-2 (CD(86)), CD(40) and CD(40L). There were no differences in the phenotype among the 3 kinds of MSCs cells. (6) The 2 kinds of culture mediums used did not markedly affect isolation and expansion of MSCs, and the biological properties of MSCs. CONCLUSIONS: (1) Human MSCs could be isolated from many kinds of human tissues, and they had no difference in their origin; (2) Human MSCs maintained good puripotentiality and self-renewal capacity. Therefore, they could meet with the need of clinical tissue engineering. (3) The negative GVHD correlated markers might result from the fact that MSCs had no HLA barrier but had broad clinical use. PMID- 14744384 TI - [Diagnostic values of serum levels of HA, PC III, C IV and LN to the liver fibrosis in children with beta-thalassemia major]. AB - OBJECTIVE: The presence of liver fibrosis in patients with beta-thalassemia major has been demonstrated to be an important negative prognostic factor. Identification of liver fibrosis in early stage would be of great value. Hyaluronic acid (HA), type III pre-collagen (PC III), collagen IV (C IV) and laminin (LN) as serum markers were widely used in the diagnosis of liver fibrosis in patients with chronic viral infections or alcoholic liver diseases. However, their values in thalassemic liver fibrosis have not been studied. This work was to determine the serum HA, PC III, C IV and LN levels in children with beta thalassemia major and evaluate the diagnostic utility. METHOD: Serum HA, PC III, C IV and LN in 49 hospitalized children with beta-thalassemia major (aged 1 - 15 years with the media age of 6.27 years) and 41 healthy children served as controls (aged 1 - 13 years with media age of 6.40 years) were detected by radioimmunoassay (RIA). Forty-five of 49 cases were performed percutaneous liver biopsies, and the histopathological fibrosis was compared with the four serum markers. The correlation and discriminate analysis were used. RESULTS: All the serum levels of HA, PC III, C IV and LN in beta-thalassemia were significantly higher than those in controls (P < 0.01). In 36 of 45 cases, the histopathology showed liver fibrosis including stage I and stage II by biopsies with a positive rate of 80%. The serum levels of four markers increased successively with the aggravation of liver fibrosis from stage 0 to stage II, and significant correlation was observed between the level of HA or PC III and the stage of fibrosis (HA, r = 0.379, P = 0.017; PC III, r = 0.455, P = 0.04). While there was no difference between the level of C IV or LN and fibrosis (C IV, r = 0.312, P = 0.053; LN, r = 0.310, P = 0.055). Using discriminate analysis, the discriminate function of co-detection of the four markers for the diagnosis of fibrosis was 0.002 HA + 0.003 PC III + 0.002 C IV + 0.006 LN-1.859, which had a sensitivity of 93.88%, specificity of 68.29%, predictive value of positive test and negative test of 77.97% and 90.32%, respectively. Moreover, there was a significant correlation between the serum level of HA or PC III and the liver iron concentration (HA, r = 0.318, P = 0.035; PC III, r = 0.305, P = 0.044). CONCLUSION: The results suggest that, in beta-thalassemia major with chronic liver disease, HA and PC III showed more practical value in diagnosing liver fibrosis than the levels of C IV and LN. The combination of the four serum markers could improve the accuracy and reliability of the diagnosis. A validation study is necessary before introducing into the prediction function during the clinical practice. PMID- 14744386 TI - [The recommend of diagnosis and treatment for childhood HIV infection and AIDS]. PMID- 14744388 TI - [Effects of intrauterine acute ischemia on epidermal growth factor and its receptor of the fetal kidney in rats]. PMID- 14744387 TI - [The effects of endotoxin on the Th1/Th2 cells and immune modulation of Astragalus membranaceus]. PMID- 14744389 TI - [The distribution of 131I-anti-CD45 antibody in mice]. PMID- 14744390 TI - [The pathological findings and inflammatory responses in the lung tissue of neonatal rats following lipopolysaccharide treatment]. PMID- 14744391 TI - [ARDS in children: a prospective clinical study in four children's hospitals in Shanghai]. PMID- 14744392 TI - [The diagnosis and differential diagnosis of three suspect SARS cases]. PMID- 14744393 TI - [Analysis of apnea in 229 cases of very low birth weight infants]. PMID- 14744394 TI - [The primary study on immunologic status of umbilical blood]. PMID- 14744395 TI - [The clinic diagnosis and analysis of two cases of congenital pulmonary dysplasia]. PMID- 14744397 TI - [Summary of national conference on pediatric infectious diseases]. PMID- 14744396 TI - [Clinical observation of Rituximab in a CD20+B non-Hodgkin's lymphoma]. PMID- 14744398 TI - [A case of treatment of convulsion caused by poison of tetramethylenedisulfotetramine with medazolam]. PMID- 14744399 TI - [Combination of novel therapy for respiratory failure]. PMID- 14744400 TI - [The role of the host defense response in ARDS and sepsis and its correlations to glucocorticoid treatment]. PMID- 14744401 TI - [Progresses in studies on SARS-associated coronavirus]. PMID- 14744402 TI - [A case of restrictive cardiomyopathy]. PMID- 14744403 TI - Hand dermatitis: a review of clinical features, therapeutic options, and long term outcomes. AB - Hand dermatitis is a common skin condition that may be chronic, debilitating, and costly for patients, insurers, and employers. The epidemiology, clinical features, occupational issues, and long-term outcomes of hand dermatitis are summarized in this review. Therapeutic options are also discussed in detail, focusing on treatment of recalcitrant hand dermatitis. PMID- 14744404 TI - Patch testing with thimerosal in a Canadian center: an 11-year experience. AB - BACKGROUND: Thimerosal has been used for decades as an antiseptic, a disinfectant, and a preservative in various consumer products. It is also a notoriously frequent contact allergen, but the significance of positive patch test reactions has been debated over the past decade. OBJECTIVE: The aim of this study was to assess the prevalence and relevance of positive patch-test reactions to thimerosal in a large Canadian center and to compare our results with those of previous publications. METHODS: A retrospective analysis of data collected over the course of 11 years in the contact dermatitis clinic of a university hospital was performed. RESULTS: Of 2252 subjects patch-tested for thimerosal, positive reactions were observed in 102 individuals (4.53%), placing thimerosal as the fifth most common allergen in our practice. The sensitization rate was equivalent between the patients who underwent aimed patch testing and those who were routinely tested for reaction to thimerosal when this allergen was included in our standard patch-testing series 8 years into the study. There was an increased incidence of thimerosal sensitization in chemists and laboratory technicians, as well as in health care workers, including medical doctors, registered nurses, dentists, and dental assistants. While eight reactions were deemed possibly relevant, in none of the cases was it possible to definitely establish the relevance of positive reactions to thimerosal, either because the presence of this allergen could not be verified in patients' products or because patch testing with these products gave negative results. CONCLUSION: In agreement with previous reports, we conclude that even though positive reactions to thimerosal are frequent, very few seem to be clinically relevant. PMID- 14744405 TI - A cross-sectional survey of complementary and alternative medicine use in patients with atopic dermatitis. AB - BACKGROUND: Studies from Europe reveal that a large percentage of patients with atopic dermatitis use some form of complementary and alternative medicine for the treatment of their skin disease. There are no studies from the United States that examine the prevalence and types of complementary and alternative medicine use among patients with atopic dermatitis. OBJECTIVE: The goal of this study was to determine the prevalence of complementary and alternative medicine use and the patterns of this use among atopic dermatitis patients referred to a university clinic. METHODS: Using a self-administered questionnaire, investigators performed a cross-sectional survey of 70 consecutive patients diagnosed with atopic dermatitis. RESULTS: Of the patients surveyed, 50.4% used some form of complementary and alternative medicine for the management of their skin disease. Common motivating factors were dissatisfaction with conventional treatment and frustration with the chronic nature of the condition. Vitamin supplementation and herbal creams were the most common alternative treatments used. CONCLUSION: The significant percentage of patients seeking alternative care for atopic dermatitis underscores the need for physicians to be familiar with alternative therapies for skin disease. PMID- 14744407 TI - Late patch-test reactions to budesonide need not be a sign of sensitization induced by the test procedure. AB - Patch-test sensitization may be suspected when a flare-up is seen at a test site at least 10 days after the test application. We describe two patients allergic to budesonide who were only diagnosed at patch-test readings on day 10 and day 13, respectively. The patients were patch-tested to a standard series including two corticosteroid mixes and their three constituents in petrolatum, namely, budesonide, tixocortol pivalate, and hydrocortisone-17-butyrate, at differing concentrations. In both patients, positive reactions to the mixes were seen on day 6 or day 7, but positive reactions to the budesonide preparations at 0.1% and 0.002% were not seen on the first or second ordinary reading day, day 3 or 4, and day 6 or 7, but no positive reactions to both budesonide preparations were seen on day 10 and on day 13, respectively. Not all late patch-test reactions represent patch-test sensitization, at least not to budesonide. PMID- 14744406 TI - Multiple patch-test reactions: a pilot evaluation of a combination approach to visualize patterns of multiple sensitivity in patch-test databases and a proposal for a multiple sensitivity index. AB - BACKGROUND: The variety of patterns of multiple sensitivity in patch-test data remains poorly defined. Studies addressing this topic have primarily concerned the occurrence of pairs of allergens, and some reports hypothesize a predisposing factor that influences the individual sensitivity of patients to multiple reactions. OBJECTIVE: The aim of this study was to address this topic by reanalyzing a matched data set from two patch-test units in Cleveland, OH, and one unit in Cologne, Germany, focusing on multiple reactions to identical allergens of the standard screening trays over the same 4-year period. METHODS: Based on the statistical FREQ procedure (SAS Institute Inc., Cary, NC), we propose a program for addressing the difficulties in computing and visualizing patterns of multiple sensitivity. Additionally, we propose the "multiple sensitivity index" (MSI) as an absolute measure for characterizing the occurrence of an allergen with others on a selected panel. RESULTS: For the seventeen allergens examined, 131072 possible combinations were evaluated in a total of 2881 patients. Of patients tested, 12.4% had multiple positive patch-test reactions to two to seven allergens. However, because of the small number of patients with the crucial number of possible combinations, no cluster patterns were evident in the three- to seven-allergen combinations. Pairs of allergens most frequently observed were nickel sulfate and potassium dichromate (n = 23), formaldehyde and quaternium-15 (n = 18), and nickel sulfate and formaldehyde (n = 13). We found that nickel sulfate, once again the most frequent sensitizer, occurred in the majority of noncombined cases (MSI = -0.280). CONCLUSION: Larger patch-test databases require evaluation to obtain further evidence of cluster patterns of multiple sensitivity and to validate the MSI. PMID- 14744408 TI - Permanent-wave dermatitis: contact allergy to cysteamine hydrochloride. AB - Cysteamine hydrochloride (CHC) is a newly recognized sensitizer found in permanent-wave solutions. We report the case of a hairdresser who was found to be allergic to CHC. Our allergic patient was patch-tested to various chemicals found in permanent-wave solutions, including CHC (1.0% in petrolatum). Patch-test reactions were positive to CHC, glyceryl thioglycolate, diglyceryl thioglycolate, p-phenylenediamine (PPD), and PPD through a piece of latex glove. Sixty-four controls to CHC (1.0% in petrolatum) had negative results. Household-weight latex gloves were protective against CHC allergy. Persons with CHC-waved hair were not allergic. CHC contained in "neutral" permanent-wave preparations has been used in American beauty salons since 1993. We briefly discuss the introduction and significance of CHC in permanent waves. PMID- 14744409 TI - Photocontact allergy to diallyl disulfide. AB - Although phototoxic reactions to plants are common, photoallergic contact dermatitis to plants and plant products rarely occurs. Our objective was to review the importance of including diallyl disulfide in the evaluation of patients with suspected photosensitivity. Phototests for ultraviolet B, ultraviolet A, and visible light as well as patch tests and photopatch tests for 49 allergens from the New York University Skin and Cancer Unit Photopatch Test Series were performed. Three patients had positive photopatch-test results to diallyl disulfide, which is the allergen in garlic. The authors conclude that although photocontact allergy to diallyl disulfide is rare, this allergen should be included in photopatch-test series. PMID- 14744410 TI - Occupational allergic contact dermatitis from xanthates and carbamates in mining processes. AB - The objective of this article is to describe allergic contact dermatitis from sodium isopropyl xanthate, potassium amyl xanthate, and carbamates in a geotechnician, to discuss possible cross-reactions, and to report the widespread use of carbamates and mercaptobenzothiazole in mining processes. PMID- 14744411 TI - Contact dermatitis to a limb prosthesis. AB - PROSTHESIS USERS commonly develop various skin problems on the residual limb, directly under the prosthetic device when the device is in direct contact with the skin. Prolonged occlusion and humidity increase the likelihood of developing contact sensitivity to moisturizing creams, medicaments, and materials in the prosthesis itself.1 Allergic contact dermatitis to various prosthetic design materials is uncommon, and the relevance of positive patch-test results to chemicals present in prostheses may be difficult to establish. Most reports of allergic contact dermatitis to prostheses have been concerned with surgical amputees and not congenital amputees. We report a 5-year-old boy with a transverse partial hemimelia who developed allergic contact dermatitis to an adhesive used in his myoelectric prosthesis. Unlike most prostheses, myoelectric prostheses are worn directly against the skin, for surface electromyography electrode contact. PMID- 14744412 TI - Occupational contact dermatitis caused by rhodium. PMID- 14744413 TI - Allergic contact dermatitis from colored surgical suture material: contact allergy to epsilon-caprolactam and acid blue 158. PMID- 14744414 TI - Bacitracin: allergen of the year. PMID- 14744415 TI - Patch-testing practices of American Contact Dermatitis Society members: a cross sectional survey. AB - BACKGROUND: Patch testing is an example of how medicine blends art and science, resulting in differences among practitioners in methods of history taking, application of patch tests, and interpretation of results. OBJECTIVE: To evaluate the "art" of patch testing (specifically, the prevalence of Thin-Layer Rapid Use Epicutaneous (T.R.U.E.) TEST use, photopatch testing, patch testing to dust mites, and immediate testing for latex allergy) as practiced by American Contact Dermatitis Society (ACDS) members and to compare the results of this survey with those of similar studies among United States dermatologists and allergists. METHODS: A cross-sectional survey of all 322 members of the ACDS. RESULTS: The response rate was 46%. The largest proportions of respondents were male, had practiced for 11 to 20 years, were located in the northeast United States, and were in private practice. Forty percent of respondents used T.R.U.E. TEST, and 65% performed photopatch testing. Seventy percent of respondents evaluated patients for latex allergy, and 15% patch-tested to dust mites. CONCLUSIONS: As expected, respondents reported significantly higher prevalences of patch testing, photopatch testing, and evaluation for latex allergy than did United States dermatologists generally. Respondents reported significantly lower prevalences of single patch-test readings and use of T.R.U.E. TEST than did either general United States dermatologists or general United States allergists. PMID- 14744416 TI - Shoe contact dermatitis in Israel. AB - BACKGROUND: Patch testing with the shoe series may be useful for the diagnosis of shoe contact dermatitis. OBJECTIVE: The aim of the study was to verify the usefulness of the shoe series and the association of fungal infection with positive results in the shoe series. METHODS: A total of 140 patients with suspected shoe contact dermatitis underwent patch testing with the standard series and shoe series; 41 underwent mycologic examination. RESULTS: Positive results in the shoe series were noted in 58 patients (41.4%). The most common allergens were potassium dichromate (27.2%), nickel sulfate (11.9%), Kathon CG (10.9%), and 4-tert-butylphenol formaldehyde resin (8.7%). Of the 41 patients with suspected tinea pedis, 13 (32%) had positive mycologic findings and 9 (69.2%) also had a positive response to the shoe series. Of the patients with a negative result on mycologic examination, only 10 (35.7%) had a positive result in the shoe series. CONCLUSION: Our study indicates that all patients with foot dermatitis should undergo patch testing with the shoe series. Positive results in the shoe series seem to be more prevalent in patients with a fungal infection. PMID- 14744417 TI - Pilot evaluation of a convenient and cost-effective method of patch testing. AB - OBJECTIVE: The primary objective of this double-blind randomized, pilot study was to evaluate whether previously prepared stored patches may be as effective as freshly prepared patches are for detecting contact allergy. METHODS: Two sets of 62 nonaqueous allergens were applied to participants' upper backs. One set was freshly prepared, and the second set was previously prepared and stored for an average of 6 weeks. Right or left back location for each set was randomly assigned. The primary outcome assessment compared the proportions of fresh and strong reactions in pairs with complete discordance (negative on one side and 1+, 2+, or 3+ on the other side). RESULTS: In 71 patients enrolled, there were 169 positive reactions. Concordance (any combination of a 1+, 2+, or 3+ reaction on both sides) was found in 63% of pairs (95% CI: 55.4, 70.0). Of the 25 pairs with complete discordance, there was no statistically significant difference between the rate of positive reactions for fresh and stored patches. CONCLUSION: Pilot evidence suggests that stored patches may be effective in detecting the majority of allergic contact reactions. Larger studies are needed to evaluate antigen specific effects as well as effects of varying lengths of antigen storage. PMID- 14744418 TI - Occupational contact dermatitis II: risk assessment and prognosis. AB - Contact dermatitis is a common and important condition in the occupational setting. In a companion paper, I describe changes in the incidence of recorded occupational skin disease from 1972 to 1999. Despite substantial improvements in workplace hygiene, the incidence of occupational skin disease remains half of that recorded in 1972. In the companion paper, it is argued that a more sophisticated approach to prevention and management may be required to substantially reduce the burden of occupational skin disease further. In this paper, I address the present state of risk assessment (including components of hazard identification, measuring or estimating dermal exposure, percutaneous absorption, dose-response relationships, and risk characterization) and the poor prognosis of serious occupational contact dermatitis, with a view to potential improvements in practice and outcomes. PMID- 14744419 TI - Occupational airborne contact dermatitis. PMID- 14744420 TI - Contact urticaria, allergic contact dermatitis, and photoallergic contact dermatitis from oxybenzone. AB - There is little literature regarding conventional patch tests and photopatch tests to oxybenzone resulting in both immediate- and delayed-type hypersensitivity reactions. A patient was patch-tested and photopatch-tested to various sunscreen chemicals. Both immediate- and delayed-type hypersensitivity reactions were observed with oxybenzone. The positive patch tests were also photoaccentuated. Oxybenzone, a common sunscreen allergen, can result in both contact urticaria and delayed-type hypersensitivity on both conventional patch testing and photopatch testing. Allergic contact dermatitis to sunscreen chemicals has traditionally included contact urticaria, allergic contact dermatitis, and photoallergic contact dermatitis. Due to the recognition of p aminobenzoic acid (PABA) and its esters as sensitizers, the presence of benzophenones in "PABA-free" sunscreens has become more prevalent, especially in sunscreens with a sun protection factor (SPF) greater than 8. In our patient, immediate- and delayed-type hypersensitivity reactions were seen to oxybenzone (2 hydroxy-4-methoxybenzophenone, 2-benzoyl-5-methoxyphenol, benzophenone-3, Eusolex 4360, Escalol 567, EUSORB 228, Spectra-Sorb UV-9, Uvinul M-40) upon conventional patch testing and photopatch testing. PMID- 14744421 TI - Intolerance to topical products may be due to dermographism. AB - Patients with reactions to topical products may be eliciting a physical urticaria, dermographism, by rubbing. These reactions may be misinterpreted as allergic, and three cases demonstrating this phenomenon were reviewed. All patients with reactions to topical products due to dermographism improved with counseling and antihistamine therapy. Repeat open application testing confirmed the safety of previously suspect medications in two of the three cases, preventing unnecessary changes in the medication regimens and inaccurate diagnoses of medication allergy. We observe that intolerance to topical medications due to dermographism can usually be managed without extensive testing or treatment. PMID- 14744422 TI - Telangiectatic erythematous cutaneous reaction to an implantable cardioverter defibrillator. AB - A 76-year-old man developed progressive blanchable reticulate erythema at the implantation site of his third implantable cardioverter defibrillator (ICD). Despite reports of allergic contact dermatitis to implantable devices, extensive routine patch testing in our patient failed to identify a responsible allergen. The clinical presentation and pathologic evidence were consistent with reticular telangiectatic erythema that has been reported overlying pacemakers and, in the English literature recently, overlying ICDs in two patients. Reticular telangiectatic erythema is a benign reactive condition that is possibly due to a combination of aberrant local circulatory and physical forces such as heat, electric fields, or magnetic fields. PMID- 14744423 TI - Allergic contact dermatitis from a paper mill slimicide containing 2-bromo-4' hydroxyacetophenone. AB - Slimicides are biocidal products used in paper mills to inhibit the proliferation of slime-forming microorganisms that would otherwise spoil the paper products. A laboratory technician working at a paper mill had recurring dermatitis related to contact with the slimicide Busan 1130. We report the first case of allergic contact dermatitis from the slimicide Busan 1130. Diagnostic patch testing was performed with solutions of Busan 1130 and its active ingredient, 2-bromo-4' hydroxyacetophenone (BHAP). Twenty-five controls were also tested. The patient showed a ++ reaction to 0.1% Busan 1130 aqueous solution and 0.01% BHAP in ethanol. All controls were negative. The patient had recurrent allergic contact dermatitis from exposure to BHAP contained in the slimicide Busan 1130. PMID- 14744424 TI - Stinging nettle dermatitis. AB - The stinging nettle (Urtica dioica) is a common weed that can cause a wide range of cutaneous reactions. Contact with the hairs or spines on the stems and leaves of the stinging nettle causes the release of several biologically active substances. The released chemicals act to cause itching, dermatitis, and urticaria within moments of contact. Extracts from the stinging nettle may provide therapeutic value for some inflammatory medical conditions. There is no standard treatment for stinging nettle dermatitis. PMID- 14744427 TI - Seeing is believing: the bicoid morphogen gradient matures. AB - Although Cell has a long history of publishing some of the most significant advances in developmental biology, the back to back papers by Driever and Nusslein-Volhard on the role of the Bicoid gradient in patterning the Drosophila embryo stand out as the first molecular demonstration of two of the longest standing concepts of the field, namely localized cytoplasmic determinants and morphogen gradients. Here we discuss the impact of this ground-breaking work and review recent results on bicoid mRNA localization and the dual role of Bicoid as a transcription and translation factor. PMID- 14744428 TI - The mechanisms of vesicle budding and fusion. AB - Genetic and biochemical analyses of the secretory pathway have produced a detailed picture of the molecular mechanisms involved in selective cargo transport between organelles. This transport occurs by means of vesicular intermediates that bud from a donor compartment and fuse with an acceptor compartment. Vesicle budding and cargo selection are mediated by protein coats, while vesicle targeting and fusion depend on a machinery that includes the SNARE proteins. Precise regulation of these two aspects of vesicular transport ensures efficient cargo transfer while preserving organelle identity. PMID- 14744429 TI - Rho and Rac take center stage. AB - Many features of cell behavior are regulated by Rho family GTPases, but the most profound effects of these proteins are on the actin cytoskeleton and it was these that first drew attention to this family of signaling proteins. Focusing on Rho and Rac, we will discuss how their effectors regulate the actin cytoskeleton. We will describe how the activity of Rho proteins is regulated downstream from growth factor receptors and cell adhesion molecules by guanine nucleotide exchange factors and GTPase activating proteins. Additionally, we will discuss how there is signaling crosstalk between family members and how various bacterial pathogens have developed strategies to manipulate Rho protein activity so as to enhance their own survival. PMID- 14744430 TI - Back to the future with ubiquitin. AB - Two papers published in 1984 by the Varshavsky laboratory revealed that the ubiquitin/proteasome pathway is the principal system for degradation of short lived proteins in mammalian cells, setting the stage for future demonstrations of this pathway's many regulatory roles. This perspective discusses the impact of those papers and highlights some of the subsequent insights that have led to our current appreciation of the breadth of ubiquitin-mediated signaling. PMID- 14744431 TI - Specificity in signal transduction: from phosphotyrosine-SH2 domain interactions to complex cellular systems. AB - Over the last two decades, a new and unifying concept of cellular organization has emerged in which modular protein-protein interactions provide an underlying framework through which signaling pathways are assembled and controlled. In this scheme, posttranslational modifications such as phosphorylation commonly exert their biological effects by regulating molecular interactions, exemplified by the ability of phosphotyrosine sites to bind selectively to SH2 domains. Although these interactions are rather simple in isolation, they can nonetheless be exploited to generate complex cellular systems. Here, I discuss experiments that have led to this view of dynamic cellular behavior and identify some current and future areas of interest in cell signaling. PMID- 14744432 TI - Cell death: critical control points. AB - Programmed cell death is a distinct genetic and biochemical pathway essential to metazoans. An intact death pathway is required for successful embryonic development and the maintenance of normal tissue homeostasis. Apoptosis has proven to be tightly interwoven with other essential cell pathways. The identification of critical control points in the cell death pathway has yielded fundamental insights for basic biology, as well as provided rational targets for new therapeutics. PMID- 14744433 TI - Recycling the cell cycle: cyclins revisited. AB - I discuss advances in the cell cycle in the 21 years since cyclin was discovered. The surprising redundancy amongst the classical cyclins (A, B, and E) and cyclin dependent kinases (Cdk1 and Cdk2) show that the important differences between these proteins are when and where they are expressed rather than the proteins they phosphorylate. Although the broad principles of the cell cycle oscillator are widely accepted, we are surprisingly ignorant of its detailed mechanism. This is especially true of the anaphase promoting complex (APC), the machine that triggers chromosome segregation and the exit of mitosis by targeting securin and mitotic cyclins for destruction. I discuss how a cyclin/Cdk-based engine could have evolved to assume control of the cell cycle from other, older protein kinases. PMID- 14744434 TI - Principles of tumor suppression. AB - Molecular genetic studies of familial cancer syndromes identified and defined the recessive nature of tumor suppressor genes and resolved the paradox of why tumors arising in such families exhibited an autosomally dominant pattern of inheritance. Subsequent characterization of tumor suppressor proteins revealed their widespread involvement in sporadic cancers and pinpointed key mechanisms that protect animals against tumor development. We now recognize that tumor suppressor genes regulate diverse cellular activities, including cell cycle checkpoint responses, detection and repair of DNA damage, protein ubiquitination and degradation, mitogenic signaling, cell specification, differentiation and migration, and tumor angiogenesis. Their study has become a centerpiece of contemporary cancer research. PMID- 14744435 TI - Regulation of RNA polymerase II transcription by sequence-specific DNA binding factors. AB - In eukaryotes, transcription of the diverse array of tens of thousands of protein coding genes is carried out by RNA polymerase II. The control of this process is predominantly mediated by a network of thousands of sequence-specific DNA binding transcription factors that interpret the genetic regulatory information, such as in transcriptional enhancers and promoters, and transmit the appropriate response to the RNA polymerase II transcriptional machinery. This review will describe some early advances in the discovery and characterization of the sequence specific DNA binding transcription factors as well as some of the properties of these regulatory proteins. PMID- 14744436 TI - The nucleosome: from genomic organization to genomic regulation. AB - Electron micrographs first confirmed that the eukaryotic genome is organized into repeating disk-shaped nucleosomal units composed of histones and their associated DNA. Those images made clear the function of the nucleosome in packaging and condensing the genome. Today, nucleosomes are recognized as highly dynamic units through which the eukaryotic genome can be regulated with epigenetically heritable consequences. This review focuses on the conserved protein structures that mobilize and remodel nucleosomes and specifically mark and recognize their histone and DNA components. These events directly impact DNA transcription, replication, recombination, and repair. PMID- 14744437 TI - Beginning to understand the end of the chromosome. AB - In their 1985 Cell paper, Greider and Blackburn announced the discovery of an enzyme that extended the DNA at chromosome telomeres in the ciliate, Tetrahymena. Since then, there has been an explosion of knowledge about both the RNA and protein subunits of this unusual ribonucleoprotein enzyme in organisms ranging from the ciliates to yeast to humans. The regulation of telomerase is now understood to take place both at the level of synthesis of the enzyme and via the state of its substrate, the telomere itself. The roles of telomerase in both cellular immortality and cancer are vibrant areas of current research. PMID- 14744438 TI - MicroRNAs: genomics, biogenesis, mechanism, and function. AB - MicroRNAs (miRNAs) are endogenous approximately 22 nt RNAs that can play important regulatory roles in animals and plants by targeting mRNAs for cleavage or translational repression. Although they escaped notice until relatively recently, miRNAs comprise one of the more abundant classes of gene regulatory molecules in multicellular organisms and likely influence the output of many protein-coding genes. PMID- 14744439 TI - Unraveling V(D)J recombination; insights into gene regulation. AB - V(D)J recombination assembles antigen receptor genes from component gene segments. We review findings that have shaped our current understanding of this remarkable mechanism, with a focus on two major reports--the first detailed comparison of germline and rearranged antigen receptor loci and the discovery of the recombination activating gene-1. PMID- 14744440 TI - Mammalian prion biology: one century of evolving concepts. AB - Prions have been responsible for an entire century of tragic episodes. Fifty years ago, kuru decimated the population of Papua New Guinea. Then, iatrogenic transmission of prions caused more than 250 cases of Creutzfeldt-Jakob disease. More recently, transmission of bovine spongiform encephalopathy to humans caused a widespread health scare. On the other hand, the biology of prions represents a fascinating and poorly understood phenomenon, which may account for more than just diseases and may represent a fundamental mechanism of crosstalk between proteins. The two decades since Stanley Prusiner's formulation of the protein only hypothesis have witnessed spectacular advances, and yet some of the most basic questions in prion science have remained unanswered. PMID- 14744441 TI - After the holy grail: establishing a molecular basis for Mammalian olfaction. AB - The quest to identify mammalian odorant receptors was a triumph of molecular biology. The characterization of these molecules has provided extraordinary insight into the strategy used by one neuronal system to organize sensory structures and code complex information. The odorant receptor genes have also served as powerful tools in understanding genomic organization and gene regulation. PMID- 14744442 TI - Obesity wars: molecular progress confronts an expanding epidemic. AB - The worldwide prevalence of obesity is increasing at an alarming rate, with major adverse consequences for human health. This "obesity epidemic" is paralleled by a rapid and substantive increase in our understanding of molecular pathways and physiologic systems underlying the regulation of energy balance. While efforts to address the environmental factors that are responsible for the recent "epidemic" must continue, new molecular and physiologic insights into this system offer exciting possibilities for future development of successful therapies. PMID- 14744443 TI - Autoclave method for rapid preparation of bacterial PCR-template DNA. AB - An autoclave method for preparing bacterial DNA for PCR template is presented, it eliminates the use of detergents, organic solvents, and mechanical cellular disruption approaches, thereby significantly reducing processing time and costs while increasing reproducibility. Bacteria are lysed by rapid heating and depressurization in an autoclave. The lysate, cleared by microcentrifugation, was either used directly in the PCR reaction, or concentrated by ultrafiltration. This approach was compared with seven established methods of DNA template preparation from four bacterial sources which included boiling Triton X-100 and SDS, bead beating, lysozyme/proteinase K, and CTAB lysis method components. Bacteria examined were Enterococcus and Escherichia coli, a natural marine bacterial community and an Antarctic cyanobacterial-mat. DNAs were tested for their suitability as PCR templates by repetitive element random amplified polymorphic DNA (RAPD) and denaturing gradient gel electrophoresis (DGGE) analysis. The autoclave method produced PCR amplifiable template comparable or superior to the other methods, with greater reproducibility, much shorter processing time, and at a significantly lower cost. PMID- 14744444 TI - A disintegration method for direct counting of bacteria in clay-dominated sediments: dissolving silicates and subsequent fluorescent staining of bacteria. AB - The masking of bacteria by abundant microparticles of the clay and silt fraction and cell losses due to sonication hampered direct enumeration of bacteria in sediments dominated by fine sediments. These problems can be circumvented by dissolving silicate fine particles using hydrofluoric acid and subsequent staining of bacteria by DTAF. The developed disintegration method partly replaces mechanical separation of bacteria from sediment particles by chemical disintegration of the silicates. Recovery efficiency ranged from 90% to 111% for different clays and clay-dominated sediments. Especially for the analysis of fine sediments and clays, this method circumvents both strong dilution of the sediment sample and harsh sonication. The method can also therefore be used in sediments where particle abundance is several orders of magnitude higher than bacterial abundance and simple dilution would not suffice in reliably counting bacteria. PMID- 14744445 TI - Three-dimensional differentiation of photo-autotrophic biofilm constituents by multi-channel laser scanning microscopy (single-photon and two-photon excitation). AB - A simple microscopic method to three-dimensionally differentiate between various members in photo-autotrophic biofilm systems is described. By dual-channel single photon (confocal) and two-photon laser scanning microscopy, the signals in the red and far red channels as well as their combination can be simultaneously recorded. The method takes advantage of the autofluorescent signal of cyanobacteria-recorded in the red and far red channel and the autofluorescent signal of the green algae-recorded in the far red channel only. The differentiation is based on the specific pigment composition of cyanobacteria and green algae in combination with the appropriate filter settings for detection of the autofluorescent emission signals. The method allows the non-destructive, three-dimensional examination of fully hydrated interfacial microbial communities at high resolution as well as the clear separation between autofluorescent signals of cyanobacteria and green algae. Furthermore, there is a third option to record additional signals simultaneously such as nucleic acid stained bacteria, bacteria labeled with phylogenetic probes or glycoconjugates stained by using lectins. With state of the art laser scanning microscopes, even a fourth channel is available for recording yet another parameter, e.g. in the reflection (single photon only) or fluorescence (single- and two-photon) mode. Thus the approach represents a convenient tool to study multiple parameters of complex photo autotrophic biofilm systems. PMID- 14744446 TI - Using geographical information techniques to quantify the spatial structure of endolithic boring processes within sediment grains of marine stromatolites. AB - Marine stromatolites are generated through the interactions of environmental parameters and specific microbial processes. The activities of endolithic bacteria, that bore canals through calcium carbonate (CaCO(3)) sand grains (ooids) and reprecipitate the CaCO(3) as a single layer (i.e. micritic laminae) are especially important in the longer term stability of the stromatolite macrostructure. Image analysis and classification approaches have been used previously, but only seldom as a quantitative microscopic tool. Here, we develop a new approach that enables the quantification of microscale (i.e. micrometers to millimeters) spatial structure within marine stromatolites. To demonstrate our approach, images were acquired from two different layers of a stromatolite: "orange layers", where microboring of canals within ooids was relatively abundant, and "white layers" where microboring was greatly reduced or lacking. Images were then transformed into spatial maps. Computation of canal and ooid grain areas within each image was conducted and statistically compared between replicate samples from the two stromatolite layers. This allowed quantification of the areas of ooid grains that were microbored. Based on our results, we suggest that our method could be widely applicable to sedimentary environments, and other areas of fundamental research. PMID- 14744447 TI - Soil washing improves the recovery of total community DNA from polluted and high organic content sediments. AB - Treatment of soil with surfactants and chelating agents is used in bioremediation studies to desorb and solubilize contaminants to increase their bioavalability to microorganisms. In the same way that pollutants are made more bioavailable to microorganisms, the procedure can be used to remove potential interfering materials from soil prior to cell lysis and extraction of DNA from indigenous microorganisms. The effect of soil washing was evaluated by extracting DNA from sediments of an intertidal freshwater wetland contaminated with hydrocarbons and from highly contaminated marine sediments from Sydney Harbour, Nova Scotia, Canada. Sediment samples had total organic carbon (TOC) contents that varied between 0.2% and 13%. The chemical lysis technique was also examined by comparison of an ammonium acetate precipitation of proteins and humic acids with a hexadecyltrimethylammonium bromide (CTAB) incubation and phenol:chloroform extraction. In this study, the incorporation of soil washing steps facilitated the desorption of contaminants from sediment surfaces and improved the recovery of DNA of amplifiable quality from both freshwater and marine sediments. CTAB contributed only slightly to the recovery of DNA of higher quality in the most contaminated sample from Sydney Harbour and was concomitant with a decrease in DNA yield in both sediment types. The incorporation of a soil washing step prior to the extraction of DNA from polluted environments may be important to solubilize and remove contaminants when high-quality DNA is required for subsequent analyses. PMID- 14744448 TI - Type-specific enzyme immunoassay for detection of pneumococcal capsular polysaccharide antigens in nasopharyngeal specimens. AB - We developed a new competitive EIA method for the demonstration of pneumococcal capsular polysaccharides from respiratory samples. The pediatric types 4, 6B, 9V, 14, 18C, 19F and 23F were selected for this study, because these capsular polysaccharides were included in the first heptavalent pneumococcal conjugate vaccines, which were used in the Finnish Otitis Media Vaccine Trial. Sensitivity of the EIA tests for purified polysaccharide antigens varied between 5 and 100 ng/ml, depending on the type. The assays performed well in 100 nasopharyngeal samples (NPS) samples processed through an enrichment culture, with an almost 100% sensitivity compared with routine culture. The method appeared type specific, except that EIA for 6B capsule also detected 6A. The method is applicable for type-specific identification of pneumococcus in carriage studies. PMID- 14744449 TI - Identification of clinically relevant yeasts by PCR/RFLP. AB - For molecular diagnosis of fungal disease using DNA amplification procedures in the routine laboratory, choice of appropriate target structures and rapid and inexpensive identification of amplification products are important prerequisites. Most diagnostic procedures described thus far are characterized by limited applicability, considerable cost for laboratory equipment or low power of discrimination between species. This study aimed at identification of a PCR target appropriate for diagnosis of clinically relevant yeasts and an affordable procedure for characterization of the PCR products to the species level. Here, we describe a PCR-based system using amplification of intergenic spacers ITS1 and ITS2 and restriction length polymorphism of PCR products after sequence-specific enzymatic cleavage. We show the evaluation of the system for clinically relevant Candida species. The simple and inexpensive procedure should be instrumental for rapid identification of medically important yeasts. PMID- 14744450 TI - Colloidal chitin stained with Remazol Brilliant Blue R, a useful substrate to select chitinolytic microorganisms and to evaluate chitinases. AB - A simple and sensitive method based on the use of colloidal chitin stained with Remazol Brilliant Blue R (RBB) is proposed to evaluate chitinase activity. If this colloidal-stained substrate is included as a carbon source in a liquid medium, this technique allows the selection or the comparison of chitinolytic microorganisms. The colloidal substrate is proportionally solubilized and the dye released is spectrophotometrically quantified at 595 nm. The procedures used for the staining and fixing of RBB in the colloidal chitin, and a comparison with the commercial substrate chitin-azure, are presented. The influence of several physicochemical and enzymatic parameters on the release of dyes is also shown. Both stained substrates were used for studying the effect of pH, substrate concentration, temperature and time on the chitinase reaction of Bacillus thuringiensis Bt-107. PMID- 14744451 TI - Detection of Aspergillus fumigatus mycotoxins: immunogen synthesis and immunoassay development. AB - Immunological detection of secreted low molecular weight toxins represents a potentially novel means of diagnosing infection by the fungus Aspergillus fumigatus. Two such metabolites, gliotoxin and helvolic acid, were selected and conjugated to thyroglobulin for antisera generation in rabbits. Gliotoxin was initially activated using N-[p-maleimidophenyl] isocyanate (PMPI) and subsequently conjugated to S-acetyl thioglycolic acid N-hydroxysuccinimide activated thyroglobulin, whereas helvolic acid was activated with N-(3 Dimethylaminopropyl)-N'-ethylcarbodiimide (EDC) in the presence of thyroglobulin prior to immunisation. To facilitate subsequent antisera evaluation, both toxins were similarly conjugated to bovine serum albumin (BSA). Matrix-Assisted Laser Desorption Ionisation-Time Of Flight (MALDI-TOF) mass spectrometry and SDS-PAGE analysis confirmed covalent attachment of toxins to BSA in the ratios of 15 and 2.4 mol per mol BSA for gliotoxin and helvolic acid, respectively. Resultant high titer antisera were capable of detecting both BSA-conjugated toxins (inhibitory concentration (IC)(50): 4-5 microg/ml). Free toxins were also detectable by competitive immunoassay, whereby 10 microg/ml free gliotoxin (30 microM) and helvolic acid (17 microM), respectively, inhibited antibody binding to cognate toxin-BSA previously immobilised on microwells. This work confirms that sensitive and specific antisera can be raised against fungal toxins and may have an application in diagnosing fungal infection. PMID- 14744452 TI - PCR-mediated whole genome amplification of phytoplasmas. AB - A method was developed for genome analysis of phytoplasmas, bacterial plant pathogens that cannot be cultivated in vitro in cell-free media. The procedure includes a CsCl-bisbenzimide gradient buoyant centrifugation followed by polymerase chain reaction (PCR)-mediated whole genome amplification. The latter step involves digestion of the DNA by a restriction enzyme with an A/T-rich recognition sequence. Due to the different A/T content in the DNA of the pathogen and its plant host, the fragments originating from phytoplasma are shorter and are preferentially amplified in the PCR reaction. Products obtained were cloned and screened by dot-blot hybridization. Results showed that about 90% of recombinant clones appeared to harbor phytoplasma specific DNA inserts. Sequencing of randomly selected clones was carried out and comparison with the NCBI database confirmed the bacterial origin for the sequences, which have been assigned a putative function. The origin of the recombinant clones was further confirmed by the generation of specific amplicons from the phytoplasma-infected plant and not from the healthy control, using PCR primers devised from the sequences of the recombinant clones. This method could be used for genome-wide comparisons between phytoplasmas. PMID- 14744453 TI - Characterization of the susceptibility of mycobacteria in BACTEC 12B media containing PANTA that had been supplemented with ceftazidime, and characterization of the individual components of PANTA in the presence of C18 carboxypropylbetaine. AB - C(18)-carboxypropylbetaine (CB-18) specimen processing has enhanced the diagnosis of mycobacterioses by smear, culture, and nucleic acid amplification. However, toxic side effects of CB-18 in liquid culture, especially in the presence of antibiotics, have been reported. The interaction of CB-18 at 20-25 microg/ml with the individual components of the antibiotic supplement PANTA that had been fortified with ceftazidime (PANTA-caz) was characterized in BACTEC 12B cultures using four mycobacterial isolates. When the Mycobacterium tuberculosis isolate ATCC 27294 was examined CB-18 plus PANTA-caz did not significantly alter the time to-positive (i.e., time to a growth index (GI) of 15 (GI(15))), but did significantly increase the time to a GI of 500 (GI(500)) by approximately 8.5 days. This result could be attributed primarily to nalidixic acid, but also to ceftazidime to a lesser degree. Statistically significant increases in GI(15) of 12.5 days and GI(500) of 16.5 days were observed in the presence of CB-18 plus PANTA-caz with the Mycobacterium avium isolate ATCC 25291. These increases were due exclusively to trimethoprim. Statistically significant increases of approximately 2.5 and 9 days in GI(15) and GI(500), respectively, were observed with Mycobacterium kansasii ATCC 12478 in CB-18 plus PANTA-caz. The presence of nalidixic acid and ceftazidime were responsible for these alterations. When the behavior of the Mycobacterium fortuitum isolate ATCC 6841 was investigated in CB 18 plus PANTA-caz, significant increases in GI(15) of 8.5 days and GI(500) of 13 days were observed. The additive effects of nalidixic acid and azlocillin were responsible for these results. No single component of the PANTA-caz formulation was responsible for the interaction between CB-18 and PANTA-caz, although nalidixic acid contributed to these effects most often. These findings are consistent with the previous recommendation that CB-18 specimen processing follow a dilution-based format to ensure that the concentration of CB-18 carried-over into liquid media falls below 5-10 microg/ml. PMID- 14744454 TI - A microtiter fluorometric assay to detect the germination of Bacillus anthracis spores and the germination inhibitory effects of antibodies. AB - Bacillus anthracis spore germination is usually detected in vitro by alterations in spore refractility, heat resistance, and stainability. We developed a more quantitative, sensitive, and semi-automated procedure for detecting germination by using a microtiter kinetic reader for fluorescence spectrophotometry. The procedure was based on the increase in fluorescence of spores with time during their incubation in germination medium containing a fluorescent nucleic acid binding dye which stained germinated B. anthracis but not ungerminated (UG) spores. Spore germination in the presence of several germinants was characterized. Although L-alanine and inosine alone stimulated rapid germination in this assay, a medium containing optimal concentrations of L-alanine, adenosine, and casamino acids gave low background fluorescence, stimulated germination completely, and at a reasonable rate. Suspensions of heat-activated, UG spores of B. anthracis strain Ames were preincubated with antibodies (Abs) against whole spores to assess their effect on germination. Analyses of the germination data obtained revealed significant differences between spores pretreated with these Abs and those treated with non-immune sera or IgG. Germination inhibitory activity (GIA) was detected for several polyclonal rabbit anti-spore Ab preparations. These included anti-Ames strain spore antisera, IgG purified from the latter, and spore affinity-purified Abs from antisera elicited against four strains of B. anthracis. Abs elicited against UG as well as completely germinated Ames spores inhibited germination. Abs were ranked according to their GIA, and those specific for UG spores usually exhibited greater GIA. Direct binding to spores of these Abs was detected by an ELISA with whole un-germinated Ames spores. Although specific binding to spores by the anti spore Abs was shown, their titers did not correlate with their GIA levels. Current efforts are focused on identifying the spore antigens recognized by the anti-spore Abs, characterizing the role of these targeted antigens in disease pathogenesis, and evaluating the ability of specific anti-spore Abs to protect against infection with B. anthracis. PMID- 14744455 TI - Evaluation of an automated system for non-radiometric detection of Mycobacterium avium paratuberculosis in bovine feces. AB - Cultivation of Mycobacterium avium subsp. paratuberculosis (M. paratuberculosis) from feces remains the most reliable method to detect infected animals. The purpose of this study was to evaluate a broth-based automated system used for cultivation of mycobacteria such as M. tuberculosis from human hosts, for the detection of M. paratuberculosis in bovine feces. Bovine feces was spiked with tenfold serial dilutions of M. paratuberculosis (5x10(5) to 5x10(-1) organisms), then processed with a double-centrifugation technique that included disinfection prior to inoculation into broth tubes. The same pathogen dilution series was also inoculated directly into broth and broth with uninfected processed feces. All of the system signal-positive bottles were identified within 30 days, with the highest concentration of M. paratuberculosis detected by the system in as few as 8 days. The presence of the pathogen was confirmed with acid-fast staining and an IS900-based PCR assay when growth of M. paratuberculosis was indicated by the system. However, some of the signal-negative cultures inoculated with the equivalent of 0.5 organisms tested PCR-positive 56 days post-inoculation, indicating that longer culture periods may lead to detection of small quantities of the organisms. Additionally, it was indicated that the processing step had a detrimental effect on detection of the organism. Comparison of the broth- and Herrold's egg yolk medium (HEYM) solid media-based culture methods with defined check test specimens corroborated the experimental evaluation of this system, indicating that broth-based detection could provide a more rapid assay for M. paratuberculosis. These results suggest that this automated system could be used to detect this organism in bovine feces, but that new approaches to processing the feces for culture should be explored. PMID- 14744456 TI - Detection of clinically relevant antibiotic-resistance genes in municipal wastewater using real-time PCR (TaqMan). AB - Real-time PCR assays were developed for the quantifiable detection of the antibiotic-resistance genes vanA of enterococci, ampC of Enterobacteriaceae, and mecA of staphylococci in different municipal wastewater samples. Primer and probe designs for these resistance genes were constructed and optimised for application in standardised TaqMan PCR assays. Using reference strains, the linear measurement ranges of the assays were defined and covered concentration ranges of five to seven exponential values. Wastewater isolates of vancomycin-resistant enterococci (VRE) and beta-lactam-resistant Enterobacteriaceae were cultivated from municipal wastewaters in order to verify the specificity and sensitivity of the primer-probe systems. Additionally, clinical strains of staphylococci resistant to methicillin (MRSA) confirmed the applicability of the mecA-specific detection system. Total DNAs were extracted from five different wastewater treatment plants and used for direct TaqMan PCR detection of the resistance genes without prior cultivation. In municipal wastewater, the resistance gene vanA was detected in 21% of the samples, and ampC in 78%. The gene mecA was not found in municipal wastewater, but in two clinical wastewater samples. PMID- 14744457 TI - Artificial soil microcosms: a tool for studying microbial autecology under controlled conditions. AB - A novel artificial microcosm containing all the essential chemical components of soil, but with reduced heterogeneity and biological complexity, has been developed. Its utility for supporting realistic microbial populations was demonstrated and an example of how competing bacteria can be studied is illustrated. PMID- 14744458 TI - Evaluation of the fully automated Bactec MGIT 960 system for the susceptibility testing of Mycobacterium tuberculosis to first-line drugs: a multicenter study. AB - The accuracy of the Bactec MGIT 960 system for susceptibility testing of 177 clinical isolates of Mycobacterium tuberculosis to first line drugs (isoniazid, rifampicin, ethambutol and streptomycin) was compared with the agar reference method. The sensitivity, the ability to detect resistance, of the MGIT system was 100%, while the specificity, the ability to detect susceptibility, ranged from 98.6% to 100% for all drugs tested. PMID- 14744459 TI - Diffusion tensor imaging: background, potential, and utility in psychiatric research. AB - Diffusion tensor imaging is a variation of magnetic resonance imaging that measures the diffusion of water in tissues. This can help measure and quantify a tissue's orientation and structure, making it an ideal tool for examining cerebral white matter and neural fiber tracts. It is only beginning to be utilized in psychiatric research. This article reviews the theory behind diffusion tensor imaging, its potential to map fiber tracts in the brain, and its recent use in psychiatric research. PMID- 14744460 TI - Neuroimaging correlates of anxiety after pediatric traumatic brain injury. AB - BACKGROUND: Anxiety disorders are common after traumatic brain injury (TBI). Data on the neural correlates of these conditions are lacking. This study examines the relationship between brain damage, particularly to the orbitofrontal cortex (OFC) and temporal lobe, and anxiety symptoms and disorders. METHODS: Ninety-five children and adolescents were followed for one year postinjury. Preinjury and one year postinjury anxiety status were obtained from the parent. Magnetic resonance imaging was performed to evaluate brain lesions. The primary analysis used regression models to determine relationships between brain lesions and anxiety outcomes. As a secondary analysis, previously reported posttraumatic stress disorder (PTSD) data were reanalyzed using similar methods for purposes of comparison. RESULTS: The primary analysis showed that greater volume and number of OFC lesions correlated with decreased risk for anxiety, whereas lesions in other brain areas did not correlate with anxiety. Consistent with prior data, the secondary analysis showed an inverse correlation between OFC damage and PTSD; temporal lobe damage was positively correlated with PTSD. CONCLUSIONS: After pediatric TBI, greater damage to the OFC is associated with decreased risk for anxiety outcomes. Similar to adult data, these findings implicate OFC dysfunction in childhood anxiety. Temporal lobe damage did not correlate with anxiety, in contrast to the findings for PTSD. PMID- 14744461 TI - Decreased hippocampal 5-HT2A receptor binding in major depressive disorder: in vivo measurement with [18F]altanserin positron emission tomography. AB - BACKGROUND: Serotonin 5-HT(2A) receptors play an important role in the regulation of many functions that are disturbed in patients with major depressive disorder. Postmortem and positron emission tomography studies have reported both increased and decreased 5-HT(2A) receptor binding in different limbic and paralimbic regions. METHODS: We conducted a quantitative 5-HT(2A) receptor binding study using positron emission tomography and [(18)F]altanserin of four regions hypothesized to have altered levels of 5-HT(2A) receptors in major depressive disorder. Using a four-compartment model, the 5-HT(2A) receptor distribution was estimated by calculating the regional [(18)F]altanserin k(3)/k(4) ratio in which k(3) is the rate of binding to the receptor and k(4) is the rate of dissociation from the receptor. Forty-six antidepressant-free patients with major depressive disorder and 29 healthy control subjects were enrolled. RESULTS: 5-HT(2A) receptor binding in the hippocampus was reduced by 29% in depressed subjects (p =.004). In other regions, 5-HT(2A) receptor binding was decreased (averaging 15%) but not significantly. Both groups had similar age-dependent decreases in 5 HT(2A) receptors throughout all brain regions. CONCLUSIONS: Altered serotoninergic function in the hippocampus is likely involved in the disturbances of mood regulation in major depressive disorder, although the specific role of the 5-HT(2A) receptor changes is still unclear. PMID- 14744462 TI - Serotonin 5-HT1A, 5-HT1B, and 5-HT2A receptor mRNA expression in subjects with major depression, bipolar disorder, and schizophrenia. AB - BACKGROUND: Alterations of serotonin neurotransmission are implicated in both mood disorders and schizophrenia. Specific serotonin-receptor-based abnormalities in these psychiatric illnesses have been intensively studied; however, it has been difficult to draw any conclusions because of a lack of consensus. These inconsistencies have most likely arisen from the unavailability of selective ligands. METHODS: Our study used in situ hybridization to quantify 5-HT(1A), 5 HT(1B), and 5-HT(2A) mRNA levels in the hippocampus (HC) and 5-HT(1A) and 5 HT(2A) mRNA levels in the dorsolateral prefrontal cortex (DLPFC) of subjects with a history of major depression disorder (MDD), bipolar disorder (BPD), schizophrenia, and a normal comparison group (15 subjects per group). RESULTS: In the DLPFC, there is a significant decrease in 5-HT(1A) mRNA of subjects with MDD and in 5-HT(2A) mRNA of subjects with BPD. Subjects with MDD have a significant decrease in 5-HT(1A) mRNA in the HC; subjects with BPD and schizophrenia had increased 5-HT(1B) mRNA levels and a significant decrease in 5-HT(2A) mRNA levels in the hippocampal formation. CONCLUSIONS: Alterations in 5-HT(1A,) 5-HT(1B), and 5-HT(2A) mRNA levels in the brains of subjects with both mood disorders and schizophrenia add further support for hypothesis of dysregulation of the serotonergic system in these psychiatric disorders. PMID- 14744463 TI - Protein kinase A in postmortem brain of depressed suicide victims: altered expression of specific regulatory and catalytic subunits. AB - BACKGROUND: We recently reported reduced [3H]cyclic adenosine monophosphate binding and catalytic activity of protein kinase A in prefrontal cortex of depressed suicide victims. Here we examined the molecular basis of these alterations and whether these findings can be replicated in another cohort. METHODS: Prefrontal cortex from depressed suicide victims and nonpsychiatric controls were obtained from the Lenhossek Human Brain Program, Budapest and the Maryland Brain Collection Program. [3H]cyclic adenosine monophosphate binding and protein kinase A activity were determined by radioligand binding and enzymatic assay, respectively. Expression of catalytic and regulatory subunits was determined by quantitative reverse transcription polymerase chain reaction and Western blot, respectively. RESULTS: [3H]cyclic adenosine monophosphate binding and total and endogenous protein kinase A activity were significantly decreased in membrane and cytosol fractions of prefrontal cortex of depressed suicide victims from the Budapest cohort, with a similar magnitude (33%-40% reduction) as reported for the Maryland cohort. In both cohorts, selective reduction (36%-41%) in mRNA and protein expression of the regulatory RIIbeta and the catalytic Cbeta was observed. CONCLUSIONS: Our results suggest abnormalities in [3H]cyclic adenosine monophosphate binding and catalytic activity kinase A in brain of depressed suicide victims, which could be due to reduced expression of RIIbeta and Cbeta. These abnormalities in PKA may be critical in the pathophysiology of depression. PMID- 14744464 TI - Serotonin transporter and GABAA alpha 6 receptor variants are associated with neuroticism. AB - BACKGROUND: A tendency to experience negative affect, as measured by the neuroticism component of the Neuroticism, Extraversion, and Openness Personality Inventory (NEO-PI), is a trait marker for major depression. Epidemiologic studies indicate a strong genetic component, but to date few specific genetic variants have been definitively implicated. A serotonin transporter promoter polymorphism (5-HTTLPR) has been extensively studied in neuroticism and several psychiatric disorders, with inconclusive results. A GABA(A) receptor alpha6 subunit variant (Pro385Ser) has been associated with alcohol-related traits but has not been studied in neuroticism or depression. METHODS: A total of 384 subjects who completed the NEO-PI were genotyped at 5-HTTLPR and Pro385Ser. Associations between polymorphisms and both alcohol use and personality domains were tested. RESULTS: The 5-HTTLPR short allele (p =.008) and Pro385Ser Pro allele (p =.003) are associated with higher neuroticism scores. The 5-HTTLPR long allele (p =.006), but not Pro385Ser, is also associated with an increased presence of alcohol use. In addition, there is a nonsignificant suggestion of an interaction: the effect of 5-HTTLPR on neuroticism might be dependent on the Pro385Ser genotype. CONCLUSIONS: These findings support a role for the serotonin transporter and GABA(A) alpha6 subunit in depression-related traits. PMID- 14744465 TI - Low posttrauma GABA plasma levels as a predictive factor in the development of acute posttraumatic stress disorder. AB - BACKGROUND: Gamma amino-butyric acid (GABA) regulates the intensity and the duration of the central hyperadrenergic response in times of high stress and has been negatively associated with anxiety, depression, and sleep problems. We hypothesized that individuals with low plasma GABA levels may be more prone to develop posttraumatic stress disorder (PTSD) in the aftermath of trauma exposure. METHODS: To test this hypothesis, we measured plasma GABA levels in a population of 108 road traffic accident victims on arrival at a traumatology department and assessed them for PTSD 6 weeks later. RESULTS: The mean GABA level (nmol/mL) in the PTSD group (n = 55; M =.20; SD =.08) was significantly lower compared with members of the trauma-exposed group who did not develop PTSD [n = 17; M =.30; SD =.09), t(70) = 3.94, p =.0002]. CONCLUSIONS: Provided that GABA levels in the brain are genetically predetermined, our results would suggest that individuals with low plasma GABA levels are premorbidly more vulnerable to stress-related disorders such as acute PTSD. If replicated, plasma GABA levels measured in the aftermath of trauma exposure might help to identify individuals at high risk for developing PTSD. PMID- 14744466 TI - Mu-opioid receptor binding measured by [11C]carfentanil positron emission tomography is related to craving and mood in alcohol dependence. AB - BACKGROUND: The endogenous opioid system has been linked to alcohol dependence through animal and human studies. We investigated the relationship between alcohol craving and brain mu opioid receptors (mu-OR) in alcohol-dependent subjects. METHODS: Regional brain mu-OR binding potential (BP) was measured using [(11)C]carfentanil positron emission tomography in eight male alcohol-dependent subjects undergoing alcohol withdrawal and eight matched control subjects. Self reported alcohol craving, withdrawal, and mood were measured. RESULTS: Lower mu OR BP was associated with higher craving in the right dorsal lateral prefrontal cortex, the right anterior frontal cortex, and right parietal cortex. In these regions, alcoholics showed lower mean mu-OR BP compared with control subjects. Mu OR BP in four other brain regions also correlated with craving, but there were no group differences in receptor binding potential. Mu-OR BP also correlated with depressive symptoms in five brain regions, three of which were identified in the craving analyses. CONCLUSIONS: Results show a strong functional relationship between alcohol craving, mood, and mu-OR binding in specific brain regions of recently abstinent, alcohol-dependent men. PMID- 14744467 TI - Functional connectivity of the prefrontal cortex and the amygdala in posttraumatic stress disorder. AB - BACKGROUND: Persistent, intrusive re-experiencing in posttraumatic stress disorder (PTSD) is commonly construed as a failure of cingulate inhibition (i.e., extinction) over a hyperresponsive amygdala, based primarily on animal research of fear conditioning and the finding of cingulate hypoperfusion in PTSD. METHODS: We examined functional connectivity in patients with PTSD and healthy trauma survivors during repeated symptom provocation using H(2)O(15) positron emission tomography. RESULTS: Memory retrieval networks (right prefrontal cortex, hippocampus, and visual cortex) were common to both groups. Networks supporting autonomic and emotional control and preparatory motor action (amygdala, anterior cingulate, subcallosal gyrus, and premotor cortex) differed between the two groups and became progressively disparate with successive presentations of the traumatic script. Patterns of effective connectivity demonstrated the predominance of direct influences of the amygdala on visual cortex, subcallosal gyrus, and anterior cingulate in PTSD but not in control subjects. There was little evidence for failure of inhibition of cingulate or subcallosal cortex over the amygdala. CONCLUSIONS: These patterns might represent excessive influences of the amygdala over regions involved in autonomic, and higher-order visual memory processing in PTSD. The present data suggest that inferences of direct correspondence between animal studies and pathophysiology of PTSD should be made with caution. PMID- 14744468 TI - Ethanolamine and phosphoethanolamine inhibit mitochondrial function in vitro: implications for mitochondrial dysfunction hypothesis in depression and bipolar disorder. AB - BACKGROUND: A growing body of experimental evidence suggests that mitochondrial dysfunction, including alterations in phospholipid metabolism, might be involved in the pathophysiology of affective illnesses, such as depression and bipolar disorder. The purpose of this study was to determine whether the phosphomonoester phosphoethanolamine (PE) and the lipid metabolite choline (Cho), which are known to be altered in depression and bipolar disorder, and/or their precursors/metabolites, might directly affect mitochondrial bioenergetic function in vitro. METHODS: To this end, rates of oxygen consumption in freshly isolated, intact mitochondria were determined polarographically in the presence and absence of PE, Cho, ethanolamine (Etn), glycerophosphoethanolamine (GPE), and glycerophosphocholine (GPC). RESULTS: The data demonstrate that PE and Etn inhibit mitochondrial respiratory activity in a dose-dependent manner, whereas Cho, GPC, and GPE have no measurable effect on bioenergetic function. CONCLUSIONS: This reflects a specific inhibition by Etn and PE on mitochondrial function rather than a more generalized phenomenon induced by similarities in structure between the lipid metabolites. These results also suggest a possible relationship between mitochondrial dysfunction and altered phospholipid metabolism in the brains of patients with depression and bipolar disorder. PMID- 14744470 TI - Low respiratory sinus arrhythmia and prolonged psychophysiological arousal in posttraumatic stress disorder: heart rate dynamics and individual differences in arousal regulation. AB - BACKGROUND: There is extensive evidence that the parasympathetic branch of the autonomic nervous system can modulate psychophysiological arousal. To date, no studies have investigated associations between cardiac vagal tone and the time course of arousal during exposure to trauma-related stimuli in posttraumatic stress disorder (PTSD). METHODS: Thirty-one subjects, 29 with PTSD and 2 with partial PTSD, had electrocardiograms recorded during baseline and 2-minute traumatic and neutral script-driven imagery periods. Heart rate, respiratory sinus arrhythmia (RSA), and heart rate half-recovery to the trauma script were quantified, and subjects were divided into low and high baseline RSA groups. RESULTS: Across all participants, heart rate significantly increased from the neutral to the trauma script and RSA significantly decreased from baseline to trauma script (p < .05). As predicted, low RSA subjects had more prolonged heart rate increases to the trauma script than high RSA subjects (p < .001), and heart rate half-recovery was negatively correlated to baseline RSA (r = -.50, p =.005). CONCLUSIONS: This study is the first to find decreased RSA in response to a traumatic reminder and an association between low baseline RSA and sustained conditioned arousal in PTSD. Low vagal tone may account for deficient arousal and emotion regulation capacities often observed in PTSD. PMID- 14744469 TI - Immobilization stress elevates tryptophan hydroxylase mRNA and protein in the rat raphe nuclei. AB - BACKGROUND: Stress triggers adaptive and maladaptive changes in the central nervous system, including activation of the hypothalamic-pituitary-adrenal axis, and can trigger mood disorders and posttraumatic stress disorder. We examined the effect of immobilization stress (IMO) on gene expression of tryptophan hydroxylase (TPH), the rate-limiting enzyme in serotonin biosynthesis, and the role of cortisol in that response. METHODS: Regular and adrenalectomized Sprague Dawley rats were exposed to various repetitions of IMO. Tryptophan hydroxylase messenger ribonucleic acid (mRNA) was determined by competitive reverse transcriptase polymerase chain reaction, and TPH protein was examined by immunoblot and immunocytochemistry. RESULTS: Elevation of TPH mRNA by IMO was tissue-specific and dose-dependent. A single IMO elicited a threefold rise in TPH mRNA in median raphe nucleus (MRN), but repeated (3x) IMOs were needed for similar response in dorsal raphe nucleus (DRN). Repeated daily IMO, up to 7 days, triggered a robust induction (6-10-fold) in TPH mRNA, accompanied by corresponding rise in TPH protein levels in raphe nuclei but not in the pineal gland. The rise in TPH immunoreactivity was widespread throughout the DRN and MRN. Bilateral adrenalectomy did not prevent the IMO-triggered increase in TPH immunoreactive protein in the raphe nuclei. CONCLUSIONS: This study reveals adrenal glucocorticoid-independent induction of TPH gene expression in raphe nuclei in response to immobilization stress. PMID- 14744471 TI - Learning and memory in Holocaust survivors with posttraumatic stress disorder. AB - BACKGROUND: Impairments in explicit memory have been observed in Holocaust survivors with posttraumatic stress disorder. METHODS: To evaluate which memory components are preferentially affected, the California Verbal Learning Test was administered to Holocaust survivors with (n = 36) and without (n = 26) posttraumatic stress disorder, and subjects not exposed to the Holocaust (n = 40). RESULTS: Posttraumatic stress disorder subjects showed impairments in learning and short-term and delayed retention compared to nonexposed subjects; survivors without posttraumatic stress disorder did not. Impairments in learning, but not retention, were retained after controlling for intelligence quotient. Older age was associated with poorer learning and memory performance in the posttraumatic stress disorder group only. CONCLUSIONS: The most robust impairment observed in posttraumatic stress disorder was in verbal learning, which may be a risk factor for or consequence of chronic posttraumatic stress disorder. The negative association between performance and age may reflect accelerated cognitive decline in posttraumatic stress disorder. PMID- 14744472 TI - Combining norepinephrine and serotonin reuptake inhibition mechanisms for treatment of depression: a double-blind, randomized study. AB - BACKGROUND: Although several antidepressants are now available, all have limited efficacy and a delayed onset of action. The current study was undertaken as a proof of the concept that combining norepinephrine and serotonin reuptake inhibition would be more effective and act more rapidly than either drug alone. METHODS: Inpatients with nonpsychotic unipolar major depression and a Hamilton Depression Rating Scale (HAMD) score of at least 18 after 1 week of hospitalization without antidepressant medication were randomized to 6 weeks of treatment with fluoxetine (FLX) 20 mg/day, desipramine (DMI) adjusted to an adequate plasma level, or the combination of FLX 20 mg/day and DMI, given under double-blind conditions. Twenty-four-hour DMI levels were used to rapidly adjust DMI dose to achieve a therapeutic level and to anticipate the enzyme-inhibiting effects of FLX. Treatment-resistant patients were stratified. Patients were rated with the HAMD and the Montgomery-Asberg Depression Rating Scale (MADRS). RESULTS: Thirty-nine patients began treatment. One patient withdrew consent. The DMI-FLX combination was significantly more likely to result in remission on the MADRS than either FLX or DMI alone [53.8% vs. 7.1% and 0%, respectively; chi(2)(2) = 13.49, p =.001]. The advantage for combined treatment was not explained by history of treatment resistance or by drug plasma concentrations. Rapid response, at 1 or 2 weeks, was neither statistically nor meaningfully greater with combined treatment. CONCLUSIONS: This study supports the hypothesis that the combination of a noradrenergic and serotonergic agent is more likely to result in remission than either selective agent alone during a 6-week treatment period. PMID- 14744473 TI - Effectiveness of electroconvulsive therapy in community settings. AB - BACKGROUND: Clinical trials indicate that electroconvulsive therapy (ECT) is the most effective treatment for major depression, but its effectiveness in community settings has not been examined. METHODS: In a prospective, naturalistic study involving 347 patients at seven hospitals, clinical outcomes immediately after ECT and over a 24-week follow-up period were examined in relation to patient characteristics and treatment variables. RESULTS: The sites differed markedly in patient features and ECT administration but did not differ in clinical outcomes. In contrast to the 70%-90% remission rates expected with ECT, remission rates, depending on criteria, were 30.3%-46.7%. Longer episode duration, comorbid personality disorder, and schizoaffective disorder were associated with poorer outcome. Among remitters, the relapse rate during follow-up was 64.3%. Relapse was more frequent in patients with psychotic depression or comorbid Axis I or Axis II disorders. Only 23.4% of ECT nonremitters had sustained remission during follow-up. CONCLUSIONS: The remission rate with ECT in community settings is substantially less than that in clinical trials. Providers frequently end the ECT course with the view that patients have benefited fully, yet formal assessment shows significant residual symptoms. Patients who do not remit with ECT have a poor prognosis; this underscores the need to achieve maximal improvement with this modality. PMID- 14744474 TI - Regulation of the norepinephrine transporter by chronic administration of antidepressants. AB - BACKGROUND: Downregulation of serotonin transporter was observed previously after chronic treatment with selective serotonin reuptake inhibitors (SSRIs) but not selective norepinephrine reuptake inhibitors (NRIs). This study investigated if chronic treatment of rats with selective NRIs or SSRIs also affected the norepinephrine transporter (NET). METHODS: Rats were treated for 3 to 6 weeks by osmotic minipumps with either the selective NRIs, desipramine, or the SSRI paroxetine. RESULTS: [(3)H]nisoxetine binding sites as well as [(3)H]norepinephrine uptake were decreased in hippocampus and cortex after treatment with desipramine. By contrast, paroxetine-treated rats showed no alteration in either [(3)H]nisoxetine binding or [(3)H]norepinephrine uptake. NET messenger RNA levels in the locus coeruleus were unchanged by desipramine treatment. CONCLUSIONS: These results demonstrate that the marked decrease in NET density 1) is not a consequence of a decrease in gene expression; 2) was caused only by a selective NRI; and 3) was associated with a parallel decrease in norepinephrine uptake. PMID- 14744475 TI - Seasonal affective disorder and the G-protein beta-3-subunit C825T polymorphism. AB - BACKGROUND: Guanine nucleotide-binding proteins (G-proteins) have been implicated in affective disorders, with reports of altered signal transduction and G-protein levels. Association with seasonal affective disorder (SAD) has been found for the higher activity T-allele of the G-protein beta-3-subunit C825T polymorphism. METHODS: European SAD patients (n = 159) and matched controls (n = 159) were genotyped for the C825T. Seasonality and diurnal preference were investigated in subsets of the material (n = 177 and 92, respectively). RESULTS: We found no association between C825T and SAD (chi(2) =.09, p =.96) or seasonality (F = 1.76, p =.18). There was some evidence for an effect on diurnal preference but only in the control group (n = 46, t = -2.8, Bonferroni corrected p =.045). CONCLUSIONS: These results suggest that the G-protein beta-3-subunit 825 T-allele does not play a major role in susceptibility to seasonal affective disorder in the population studied. PMID- 14744476 TI - Milnacipran: a comparative analysis of human monoamine uptake and transporter binding affinity. AB - BACKGROUND: Though selective serotonin reuptake inhibitors have revolutionized the field of psychiatry with demonstrated efficacy in affective and anxiety disorders with minimal side effects, norepinephrine-serotonin reuptake inhibitors may provide efficacy similar to tricyclic antidepressants without the adverse side effects associated with tricyclic antidepressants. METHODS: The affinity and selectivity of milnacipran, duloxetine, venlafaxine, citalopram, amitriptyline, and nortriptyline were determined for the human serotonin, norepinephrine, and dopamine transporters. RESULTS: Both milnacipran and duloxetine were potent inhibitors of serotonin and norepinephrine uptake. Unlike duloxetine and venlafaxine, milnacipran appears serotonin transporter selective in binding (ratio = 2.61) and norepinephrine transporter selective in uptake (ratio =.45). CONCLUSIONS: Milnacipran's binding and uptake inhibition profile more closely resembles that of the tricyclic antidepressants than that of duloxetine. Whether these differences observed in vitro manifest themselves in vivo is not clear. PMID- 14744477 TI - White matter structure in autism: preliminary evidence from diffusion tensor imaging. AB - BACKGROUND: Individuals with autism have severe difficulties in social communication and relationships. Prior studies have suggested that abnormal connections between brain regions important for social cognition may contribute to the social deficits seen in autism. METHODS: In this study, we used diffusion tensor imaging to investigate white matter structure in seven male children and adolescents with autism and nine age-, gender-, and IQ-matched control subjects. RESULTS: Reduced fractional anisotropy (FA) values were observed in white matter adjacent to the ventromedial prefrontal cortices and in the anterior cingulate gyri as well as in the temporoparietal junctions. Additional clusters of reduced FA values were seen adjacent to the superior temporal sulcus bilaterally, in the temporal lobes approaching the amygdala bilaterally, in occipitotemporal tracts, and in the corpus callosum. CONCLUSIONS: Disruption of white matter tracts between regions implicated in social functioning may contribute to impaired social cognition in autism. PMID- 14744478 TI - Liposomes and disaccharides as carriers in spray-dried powder formulations of superoxide dismutase. AB - In this study, we aim to investigate the effects of disaccharides and liposome carriers on the activity, solid state characteristics, structural preservation, and aerosol powder performance of spray-dried superoxide dismutase (SOD) formulations. Sucrose, trehalose, and lactose were selected as stabilizing adjuvants in the spray-drying process. Dipalmitoyl phosphatidylcholine (DPPC) was the major lipid component for preparing liposomes. These SOD formulations were characterized with SOD activity assay, particle size, residual moisture content, scanning electron microscopy (SEM), electron spectroscopy for chemical analysis (ESCA), differential scanning calorimeter (DSC), and Fourier transform infrared (FT-IR) spectroscopy. We found that the inlet/outlet temperature of spray drying can be performed up to 168/122 degrees C with maintaining the activity of SOD in the SOD?DPPC?sucrose formulation for 98%. The SEM image of this formulation showed wrinkled and raisin-like appearance. Aerosol powder performance test demonstrated that this formulation exhibited excellent emitted dose (ED, 71%), aerodynamic diameter (2 microm), and respirable fraction (RF, 72%). DSC study suggested an indication of initial electrostatic stabilization of SOD by DPPC and sucrose, the following lipid perturbation by SOD, and the formation of an inclusion complex, thus minimizing the individual transition peaks of SOD and DPPC. FT-IR study showed that the major secondary structure of SOD, beta-sheet, was maintained in this formulation. The surface ESCA analysis of this formulation suggested the absence of SOD on the surface region of the powders, indicating that SOD was well surrounded and protected by DPPC and sucrose. Spray drying has been demonstrated to be a feasible process to preserve the activity of SOD in the formulation of DPPC liposomes with sucrose. PMID- 14744479 TI - Hot-melt extruded ethylcellulose cylinders containing a HPMC-Gelucire core for sustained drug delivery. AB - The objective of the study was to develop a sustained release system consisting of a hot-melt extruded ethylcellulose pipe surrounding a drug-containing hydroxypropyl methylcellulose (HPMC)-Gelucire 44/14 core, yielding a monolithic matrix system applicable in the domain of sustained drug release. The influence of HPMC substitution type and viscosity grade was investigated through dissolution testing and erosion studies. All sustained release systems showed a nearly constant drug release profile with only 40% of the drug released after 24 h. To achieve complete drug release after 24 h, the core formulation and the dimensions of the hollow pipe were modified. Changing the composition of the core did not result in the intended zero-order drug release. Shortening the length of the ethylcellulose cylinder accelerated drug release, while modifying the diameter did not affect the drug release rate. The drug dissolution profile and the release mechanism were independent of drug solubility. Increasing the drug loading caused a small increase of the drug release rate, but did not alter the release mechanism. PMID- 14744480 TI - Design of attachment type of drug delivery system by complex formation of avidin with biotinyl drug model and biotinyl saccharide. AB - Recent studies have focused on the active targeting of drug delivery by combining a homing device and antitumor drug. For this purpose, synthesis of a well designed vehicle (such as polymer/drug conjugates or nanoparticles) carrying a drug and a homing device requires many steps. We propose a new type of drug delivery system (DDS) by formation of a complex containing avidin (Av) plus biotinyl drug with a biotinyl homing device, which easily accommodates the combination of various drugs and homing devices. The targetable drug complex can be prepared by selecting an appropriate biotinyl drug derivative and a biotinyl homing device and mixing them with avidin. Fluorescent dye with 5-(and-6) carboxytetramethylrhodamine (TAMRA) was used as a drug model, and galactose (Gal) recognized by liver parenchymal cells was used as a homing device. TAMRA and galactose were attached to biotin (Bio) through a triethyleneglycol (TEG) spacer group to give Bio-TEG-TAMRA conjugate and Bio-TEG-Gal conjugate, respectively. Confocal laser scanning microscopic studies suggest that the complexes prepared by mixing Bio-TEG-Gal conjugate and fluorescein isothiocyanate (FITC)-labeled Av (feed molar ratio 4:1), and mixing Bio-TEG-Gal conjugate, Bio-TEG-TAMRA conjugate and FITC-labeled Av are internalized into the hepatoma cells through a receptor mediated endocytosis mechanism. PMID- 14744481 TI - Starch acetate as a tablet matrix for sustained drug release. AB - The aim of this study was to investigate the effect of a high degree on substitution (DS) on starch acetate (SA) and SA concentration on tablet properties. SAs with a DS of 2.6 and 3.0 were used as matrix formers with propranolol hydrochloride (PH) as a model drug. The SA-3.0 powder had better compactibility than the SA-2.6 powder. A decrease in SA concentration decreased compactibility of PH/SA blended powders when compared to neat SA powders. In general, drug release was considerably slower from SA-3.0 matrices than from SA 2.6 matrices. Also, a decrease in SA concentration increased the drug release rate. Water penetration into 80% (w/w) SA-3.0 matrices was incomplete during 24-h dissolution tests. Diffusion path length increased with time and PH was released by Fickian diffusion. However, all other PH/SA tablets were completely hydrated during dissolution tests. Macroscopic cracks were formed during dissolution, which increased area available for Fickian diffusion and resulted in slow attenuation of the drug release rate. Crack formation, not been reported earlier, must be taken into account in order to understand drug release from SA matrices. PMID- 14744482 TI - Controlled drug release from hydrogel nanoparticle networks. AB - Monodisperse nanoparticles of poly-N-isopropylacrylamide-co-allylamine (PNIPAM-co allylamine) and PNIPAM-co-acrylic acid (PNIPAM-co-AA) were synthesized. The close packed PNIPAM-co-allylamine and PNIPAM-co-AA nanoparticles were converted to three-dimensional gel networks by covalently crosslinking neighboring particles at room temperature and neutral pH using glutaric dialdehyde and adipic acid dihydrazide, respectively. Controlled release studies were conducted using dextran markers of various molecular weights as model macromolecular drugs. Release was quantified under various physical conditions, including a range of temperatures and dextran molecular weights. Dextran, entrapped in cavities in the nanoparticle network, was released with a rate regulated by their molecular weights and cavity size. No release from a conventional bulk PNIPAM gel, with high crosslinking density, was observed. The rate of release from the PNIPAM-co allylamine network was temperature-dependent, being much faster at room temperature than that at human body temperature. In contrast, release of low molecular weight dextrans from the PNIPAM-co-AA network showed a temperature independent release profile. These nanoparticle networks have several advantages over conventional bulk gels for controlling the release of high molecular weight biomolecules. PMID- 14744483 TI - Improvement in the disintegration of shellac-coated soft gelatin capsules in simulated intestinal fluid. AB - Shellac is a natural enteric polymer, which results in good gastric resistance; however, it often dissolves too slowly in intestinal fluids. The objective of this study was to improve the disintegration of shellac-coated soft gelatin capsules in simulated intestinal fluids (phosphate buffer pH 6.8) through the addition of pore-formers, such as organic acids and hydrophilic polymers, while retaining gastric resistance. The mechanical properties (% elongation at rupture, puncture strength at break and modulus at puncture), media uptake and weight loss of shellac films were determined upon exposure in 0.1 N HCl and/or phosphate buffer pH 6.8. Organic acids (e.g., sorbic acid) acted as plasticizers, they reduced the glass transition temperature of ethanol-cast shellac films. The addition of additives effectively decreased the disintegration times in phosphate buffer pH 6.8, while the behavior in 0.1 N HCl remained unchanged. In addition, the hardness and disintegration of shellac-coated soft gelatin capsules were monitored through the whole disintegration experiments. The best disintegration was achieved with sorbic acid as pore-former. Sorbic acid remained in the shellac coating at low pH, but leached in pH 6.8 buffer, thus resulting in good gastric resistance and rapid disintegration in simulated intestinal fluids. The disintegration time of ethanolic shellac-coated soft gelatin capsules decreased with increasing amount of pore-former. The slow disintegration of aqueous shellac coated soft gelatin capsules could be also improved by the addition of hydrophilic polymers, such as hydroxypropyl methylcellulose (HPMC). However, higher HPMC concentrations were required when compared to sorbic acid. PMID- 14744484 TI - Incorporation and release behavior of hydrophobic drug in functionalized poly(D,L lactide)-block-poly(ethylene oxide) micelles. AB - The poly(ethylene oxide)-poly(lactide) (PEO-PLA) block copolymers containing a small quantity of carboxylic acid in the PLA block were synthesized. The microscopic characteristics of nanoparticles with carboxylic acid content in the copolymer were analyzed, and the effect of specific interactions between the copolymer and the model drug on the drug loading capacity and the release behavior were investigated systematically. The sizes of nanoparticles prepared by a dialysis method are within the range of 30-40 nm. The nanoparticles prepared from functionalized block copolymers have a very low critical micelle concentration (CMC) value as low as approximately 10(-3) mg/ml, which indicates a good stability of the nanoparticles in spite of the presence of carboxylic acid. The drug loading efficiency of nanoparticles dramatically increased when carboxylic acid content was increased in the block copolymer. This result may be attributed to the increase of interactions between the copolymer and the drug. The release rate of the drug was much slower from nanoparticles containing higher amounts of carboxylic acid in the copolymer, which might be associated with the enhanced interaction between the carboxylic group of copolymers and the drug. These experimental results suggest that the nanoparticles prepared from functionalized PEO-PLA block copolymers could be a good candidate for an injectable drug delivery carrier. PMID- 14744485 TI - Study on accelerated evaluation system for release profiles of covered-rod type silicone formulation using indomethacin as a model drug. AB - The purpose of this study was to establish a method allowing rapid evaluation in vitro of the profiles of drug release from covered-rod type silicone formulation (CR silicone formulation), which releases drug for a prolonged period of time. Three CR silicone formulations containing indomethacin (IDM) with different release profiles were used in this study. The release of IDM was accelerated in a mixture of methanol and water (MeOH/water) compared with in phosphate-buffered saline (PBS) added by Tween 20 (PBS-based solvent). The velocity of IDM release varied depending on the composition of the MeOH/water. The change in release velocity was dependent on the solubility of IDM and the permeability of IDM through the silicone membrane. In all the tested formulations, the release rates of IDM estimated in 90% (v/v) MeOH/water were equally 14.6 times faster than those estimated in PBS-based solvent. Release of IDM from the cross-sections and lateral side evaluated by a bi-directional elution cell were accelerated in the MeOH/water in a similar degree. By introducing a common factor to shorten the time axis in all formulations, a fairly good agreement was observed between the two release profiles obtained in the accelerated MeOH/water system and the usual PBS-based solvent system. These results indicate that MeOH/water system enables to reduce the period for evaluation of profiles of drug release from CR silicone formulations in reflecting their release characteristics in usual PBS-based solvent system. PMID- 14744486 TI - Modulating drug release with cyclodextrins in hydroxypropyl methylcellulose gels and tablets. AB - This paper reports on the effect of beta-cyclodextrin (beta-CD) and hydroxypropyl beta-cyclodextrin (HP-beta-CD) on the diffusion and the release behavior of diclofenac sodium and sulphamethizole from HPMC K4M gels and matrix tablets. The gels were prepared with 0.5-2.0% polymer and different drug/CD mole ratios, and their viscosity, cloud point and drug diffusion coefficients were estimated. No differences in cloud point were observed. The viscosity of the gels strongly depended on HPMC proportions (from 0.7 to 100 mPa.s), which affected to a lesser extent the resistance to the diffusion of the drugs (D values from 60 x 10(-6) to 5 x 10(-6) cm(2)/s). The influence of CD on diffusion was particularly evident in gels prepared with polymer proportions above its entanglement concentration, 2.0% HPMC K4M. In these systems, while high drug/CD proportions enhanced the diffusivity preventing polymer/drug hydrophobic interactions, low drug/CD ratios hindered it. An excess of free CD, especially the bulky HP-beta-CD, made the diffusion of the complexes in the relatively low mesh size 2% polymer network more difficult. In the case of tablets, CD plays an additional role as dissolution rate promoter. To evaluate to what extent the balance between the increase in dissolution rate and the decrease in diffusion rate induced by CD determines drug release, matrix tablets were prepared by direct compression of 100 mg drug and 400 mg polymer/CD/lactose blends, whose composition was chosen following a simplex centroid design. A higher CD/lactose ratio significantly increased the release rate of hydrophobic drugs (sulphamethizole), but decreased the release rate of hydrophilic drugs (diclofenac sodium), indicating the predominance of a different contribution depending on the hydrophilicity of the drug. Therefore, the use of CD derivatives may be particularly useful to modulate drug release from HPMC gels and matrix tablets; the influence of these additives being dependent on the nature of the drug and on the molecular size and hydrophilic character of the CD used. PMID- 14744487 TI - Mechanism of bacitracin permeation enhancement through the skin and cellular membranes from an ethosomal carrier. AB - The main objective of the present work was to investigate the dermal and intracellular delivery of bacitracin, a model polypeptide antibiotic, from ethosomes. Bacitracin and fluorescently labeled bacitracin (FITC-Bac) ethosomes were characterized for shape, lamellarity, fluidity, size distribution and entrapment capacity by scanning electron microscopy (SEM), transmission electron microscopy (TEM), differential scanning calorimetry (DSC), dynamic light scattering (DLS) and ultracentrifugation, respectively. Confocal laser scanning microscopy (CLSM) experiments revealed that ethosomes facilitated the copenetration of antibiotic and phospholipid into cultured 3T3 Swiss albino mice fibroblasts. These results, confirmed by data obtained in fluorescent-activated cell sorting (FACS) experiments, suggest that ethosomes penetrate cellular membrane releasing the entrapped molecule within cells. Additional work was focused on skin permeation behavior of FITC-Bac from ethosomal systems in in vitro and in vivo experiments through human cadaver and rat skin, respectively. These studies demonstrated that the antibiotic peptide was delivered into deep skin layers through intercorneocyte lipid domain of stratum corneum (SC). Occlusion had no effect on the permeation profile of the drug from ethosomes in in vitro experiments. Efficient delivery of antibiotics to deep skin strata from ethosomal applications could be highly beneficial, reducing possible side effects and other drawbacks associated with systemic treatment. Furthermore, ethosomal delivery systems could be considered for the treatment of a number of dermal infections, requiring intracellular delivery of antibiotics, whereby the drug must bypass two barriers: the SC and the cell membrane. PMID- 14744488 TI - Nonlinear mixed-effects model for the dissolution assays of drugs. AB - Sustained or controlled release delivery systems can achieve predictable and reproducible drug release rates. With the aim of optimizing the bioavailability of conventional drugs with minimum side effects, new drug delivery systems continue to attract much attention. This research has been addressed to investigate in vitro metoclopramide release behavior from inert polymeric matrix tablets. Model-dependent and model-independent approaches are commonly used to investigate the release process and a fitted model results for each experimental condition. The aim of this work was to study the release characterization of the whole batch, therefore, we have developed a population model, taking into account the differences between tablets, experimental conditions, heterogeneity, and autocorrelation of residuals. The study of dissolution profiles was carried out using a mixed-effect model, which has provided a satisfactory tool for this kind of data. PMID- 14744489 TI - Bioadhesive grafted starch copolymers as platforms for peroral drug delivery: a study of theophylline release. AB - Nonirritant bioadhesive drug release systems based on starch-acrylic acid graft copolymers prepared by radiation of starch and acrylic acid mixtures with (60)Co were developed for buccal application. The release rate of theophylline (TPL), used as a model drug, depended on the ratio of starch to acrylic acid and on the presence of cations in the graft copolymers, but was practically not affected by the pH (between pH 3 and 7) of the dissolution medium nor by the type of starch used (corn, rice, or potato). Possible release mechanisms are discussed for specific conditions. In general, the release behavior of the graft copolymers was found to be non-Fickian, n value being between 0.6 and 0.96, suggesting that the release was controlled by a combination of tablet erosion and the diffusion of the drug from the swollen matrix. Incorporation of divalent cations into the graft copolymers led to a significant decrease in swelling erosion of the tablets as well as a substantial retardation of drug release. Highest work of adhesion was obtained with graft copolymers containing calcium ions as well as longer time of adhesion on dogs' gingiva. PMID- 14744490 TI - Effect of hyaluronidase and PEG chain conjugation on the biologic and antitumor activity of RNase A. AB - Subcutaneous application of bovine RNase A conjugated to HYase (bovine hyaluronidase), polyethylene glycol (PEG) and HYase+PEG resulted in a marked reduction of the width of the spermatogenic layers of the mouse testes. The number of sperms in caput epididymidis was significantly decreased in mice injected with conjugated RNase A. There was not any significant embryotoxic effect of free RNase A even conjugated with HYse, PEG and HYse+PEG. The immunogenicity, expressed in production of antibodies against free RNase A or conjugates with PEG, was very low. However, the immunogenic action of this enzyme conjugated only to HYase was much higher and produced the same immunogenicity as HYase itself. The immunogenic effect of RNase A+HYase conjugate decreased when PEG was joined to this conjugate. The inhibitory effect of RNase A conjugated to HYase, PEG and HYase+PEG on human ML-2 cells studied in vitro, was practically ineffective. On the other side, when RNase A conjugated to HYase or PEG was administered intraperitoneally into the mice bearing human melanoma, the antitumor effect was pronounced. PMID- 14744491 TI - Micellar formulations for drug delivery based on mixtures of hydrophobic and hydrophilic Pluronic block copolymers. AB - Micelles formed by Pluronic block copolymers (PBC) have been studied in multiple applications as drug delivery systems. Hydrophobic PBC form lamellar aggregates with a higher solubilization capacity than spherical micelles formed by hydrophilic PBC. However, they also have a larger size and low stability. To overcome these limitations, binary mixtures from hydrophobic PBC (L121, L101, L81, and L61) and hydrophilic PBC (F127, P105, F87, P85, and F68) were prepared. In most cases, PBC mixtures were not stable, revealing formation of large aggregates and phase separation within 1-2 day(s). However, stable aqueous dispersions of the particles were obtained upon (1). sonication of the PBC mixtures for 1 or 2 min or (2). heating at 70 degrees C for 30 min. Among all combinations, L121/F127 mixtures (1:1% weight ratio) formed stable dispersions with a small particle size. The solubilizing capacity of this system was examined using a model water-insoluble dye, Sudan (III). Mixed L121/F127 aggregates exhibited approximately 10-fold higher solubilization capacity compared to that of F127 micelles. In conclusion, stable aqueous dispersions of nanoscale size were prepared from mixtures of hydrophobic and hydrophilic PBC by using the external input of energy. The prepared mixed aggregates can efficiently incorporate hydrophobic compounds. PMID- 14744492 TI - Biosurfactant MEL-A dramatically increases gene transfection via membrane fusion. AB - Biosurfactants, which are surface-active compounds produced by microorganisms growing on water-insoluble substrates, have many biological activities. We studied here three different biosurfactants, mannosylerythritol lipid (MEL) or 4 O-[(4',6'-di-O-acetyl-2',3'-di-O-alkanoyl)-beta-D-mannopyranosyl] meso-erythritol (MEL-A), 4-O-[(6'-O-acetyl-2',3'-di-O-alkanoyl)-beta-D-mannopyranosyl] meso erythritol (MEL-B) and 4-O-[(4'-O-acetyl-2',3'-di-O-alkanoyl)-beta-D mannopyranosyl] meso-erythritol (MEL-C). MEL-A enhanced the efficiency of gene transfection by cationic liposomes, but MEL-B and MEL-C did not. We also studied the localization of FITC-conjugated antisense DNAs (15-mer oligonucleotides; phosphorothioate) in the target cells by confocal laser scanning microscopy (CLSM). The FITC-conjugated antisense oligonucleotides were temporarily on the plasma membrane of the target cells, thereafter they were transferred into the nucleus of the target cells. In the case of MEL-B and MEL-C, such localization of DNA was not observed both in the plasma membrane and in the nucleus. The results obtained by CLSM images were in good agreement with the transfection efficiency. This suggests that MEL-A induces the membrane fusion between the target cells and the cationic liposomes, accelerating the efficiency of gene transfection dramatically. PMID- 14744494 TI - New directions in cancer research 2003: technological advances in biology, drug resistance, and molecular pharmacology. AB - The 94th Annual Meeting of the American Association for Cancer Research (AACR) was held from July 11 to 14, 2003 in Washington, DC, and provided an overview of the latest developments in the field of cancer. This report provides highlights of presentations on array-based and RNA-interference technologies to study cancer biology and molecular pharmacology of anticancer drugs, mechanisms and modulation of drug resistance patterns, recent developments in the treatment of prostate cancer, and the medicinal chemistry of established and novel anticancer drugs. PMID- 14744493 TI - In vitro and in vivo evaluation of recombinant silk-elastinlike hydrogels for cancer gene therapy. AB - The objectives of this study were to evaluate: (i). the influences of hydrogel geometry, DNA molecular weight, and DNA conformation on DNA release from a silk elastinlike protein polymer (SELP) hydrogel, (ii). the bioactivity and transfection efficiency of encapsulated DNA over time in vitro, (iii). the delivery and transfection of a reporter gene in a murine model of human breast cancer in vivo, and (iv). the in vitro release and bioactivity of adenovirus containing the green fluorescent protein (gfp) gene as a marker of gene transfer. Plasmid DNA was released from SELP hydrogels in a size-dependent manner, with the average effective diffusivity ranging from 1.70+/-0.52 x 10(-12) cm(2)/s for a larger plasmid (11 kbp) to 2.55+/-0.51 x 10(-10) cm(2)/s for a smaller plasmid (2.6 kbp). Plasmid conformation also influenced the rate of release, with the rank order linear>supercoiled>open-circular. DNA retained bioactivity in vitro, after encapsulation in a SELP hydrogel for up to 28 days. Delivery of pRL-CMV from a SELP hydrogel resulted in increased transfection in a murine model of human breast cancer by 1-3 orders of magnitude, as compared to naked DNA. The release of a bioactive adenoviral vector was related to the concentration of the polymer in the hydrogel. These studies indicate that genetically engineered SELP hydrogels have potential as matrices for controlled nonviral and viral gene delivery. PMID- 14744495 TI - The importance of p53 location: nuclear or cytoplasmic zip code? AB - The regulation of p53 functions is tightly controlled through several mechanisms including p53 transcription and translation, protein stability, post translational modifications, and subcellular localization. Despite intensive study of p53, the regulation of p53 subcellular localization although important for its function is still poorly understood. The regulation of p53 localization depends on factors that influence its nuclear import and export, subnuclear localization and cytoplasmic tethering and sequestration. In this review, we will focus on various proteins and modifications that regulate the location and therefore the activity of p53. For example, MDM2 is the most important regulator of p53 nuclear export and degradation. Cytoplasmic p53 associates with the microtubule cytoskeleton and the dynein family of motor proteins; while Parc and mot2 are involved in its cytoplasmic sequestration. Finally, a portion of p53 is localized to the mitochondria as part of the non-transcriptional apoptotic response. In this review we strive to present the most recent data on how the activity of p53 is regulated by its location. PMID- 14744496 TI - p53 family members and chemoresistance in cancer: what we know and what we need to know. AB - Resistance to chemotherapy remains a major obstacle to the successful management of many human cancers. Numerous genetic and epigenetic changes in the cancer cell may contribute to drug resistance. However, with the recognition of important roles for both p53 and its more recently described paralog p73 in mediating the activity of anti-cancer drugs, there has been increasing recognition that cellular resistance to such agents can and does arise through failure of p53 family member signalling. Abrogation of function in p53 occurs through point mutations which abolish DNA binding or by dominant negative inhibition by variants of p73 lacking the N-terminal transactivation domain. Conversely, the function of full-length transactivation-competent (TA)p73 variants expressing the transactivation domain is itself subject to trans-dominant inhibition by certain p53 mutants and, in some cancers, by transcriptional silencing. The specificity of target gene activation by p53 and TAp73 is modulated by transcriptional co activators which thereby act as response modifiers. Changes in the activity/expression of co-activators may therefore cause an altered cellular response to p53/p73 activation. Despite these advances in understanding how cells respond to DNA damage in vitro, and how this is affected by molecular genetic changes which affect p53 family member signalling, the contribution of these to in vivo drug resistance has not been definitively established. Our major task now is to determine how these changes operate individually and collectively in vivo to produce the phenotype of clinical drug resistance, and how we can translate this knowledge into clinically useful strategies to improve the outcome of chemotherapy. PMID- 14744497 TI - Modulating protein kinase C (PKC) to increase the efficacy of chemotherapy: stepping into darkness. AB - The identification of molecules that promote chemotherapeutic resistance would allow rationally designed approaches to abrogate this resistance, thereby possibly improving clinical outcomes for patients with cancer. In this regard, the PKC family is attractive for targeting, because it is comprised of a family of isoforms that play key roles in multiple cellular processes and can contribute to cellular transformation. Encouraging in vitro data originally showed that approaches to modulate PKC activity through small-molecule inhibitors or genetic manipulation could affect tumor cell survival. Recently, some of these approaches have begun clinical testing. Early-stage clinical trials revealed scattered clinical responses to these agents, but the most recent clinical trials have shown that combining modulators of PKC with standard chemotherapy does not improve outcome over chemotherapy alone. In this review, we will trace the development of these approaches, and discuss possible explanations for the recent negative results. Importantly, we will suggest guidelines for the clinical evaluation of PKC modulators. PMID- 14744498 TI - DNA methylation and cancer therapy. AB - Vertebrate DNA is modified by methyl moieties at the 5'-position of cytosine rings residing in the di-nucleotide sequence CpG. Approximately 80% of CpG dinucleotide sequences are methylated. The pattern of distribution of methylated CGs is cell-type specific and correlates with gene expression programming and chromatin structure. Three kinds of seemingly contradictory aberrations in DNA methylation are observed in cancer, global hypomethylation, and regional hypermethylation and deregulated level of expression of DNA methyltransferases. It was previously proposed that the DNA methylation machinery is a candidate target for anticancer therapy. Inhibition of hypermethylation was the first therapeutic target. However, recent data suggests that inhibition of DNA methylation might have untoward effects such as induction of genes involved in metastasis. This review discusses the relative role of the three levels of alteration in the DNA methylation in cancer, proposes a unified hypothesis on the relative roles of increased DNA methyltransferase as well as the coexistence of hypo -and hyper- methylation in cancer and its possible implications on anticancer therapy. PMID- 14744499 TI - Mechanism of action of 2-methoxyestradiol: new developments. AB - 2-Methoxyestradiol (2ME2) is an endogenous metabolite of estrogen that has both antiangiogenic and antitumor effects. In preclinical models, 2ME2 showed promising activity that led to its clinical development as an orally active, small-molecule inhibitor of angiogenesis. Initial results suggest that 2ME2 is well tolerated and several Phase I and II clinical trials are evaluating 2ME2 in multiple tumor types. While many studies over the past 10 years have increased our understanding of how 2ME2 exerts its pleiotropic effects, its molecular mechanisms of action are not yet clear. Recent data have shown that 2ME2 inhibits HIF-1alpha, a key angiogenic transcription factor. The ability of 2ME2 to inhibit HIF-1alpha correlates with its microtubule-depolymerizing effects. The extrinsic and intrinsic pathways of apoptosis and reactive oxygen species are involved in apoptosis initiated by 2ME2; the relative contribution of each pathway appears to vary depending on the cell type. This review focuses on papers published within the past 2 years up to September 2003 that provide significant new insights into how 2ME2 exerts its diverse effects. PMID- 14744500 TI - Selective targeting of cancer cells using synthetic peptides. AB - To establish efficient and reliable therapeutic delivery into cancer cells, a number of delivery agents and concepts have been investigated in the recent years. Among many improvements in targeted and controlled delivery of therapeutics, cell-targeting peptides have emerged as the most valuable non immunogenic approach to target cancer cells. Peptides can be incorporated into multicomponent gene-delivery complexes for cell-specific targeting. In contrast to larger molecules such as monoclonal antibodies, peptides have an excellent tumor penetration, which make them ideal carriers of therapeutics to the site of primary tumor and the distant metastatic sites. Here we give an update on the progress made during the last two years on the identification and potential of specific synthetic tumor targeting peptides. PMID- 14744501 TI - Collaborative care management for a pregnant woman with an ostomy. AB - Pregnant women with an ostomy face many challenges during the prenatal period. Although these challenges may be present, pregnancy in the ostomate is presented as a continuum of health and wellness in this article. Collaborative care professionals, especially nurses and midwives, are often the most accessible professional sources for health care information, but current research and information on this topic is sparse. Ostomy-related problems that may occur during pregnancy, include: intestinal obstruction, peristomal hernia, pouching problems, including the effect of nausea and vomiting on pouch hydration, stomal retraction, stenosis, or laceration, and stomal prolapse. Three types of ostomies are discussed: continent ileostomies, urostomies and colostomies. A description of the normal characteristic appearance of each is also included. Collaborative care from pre-conception through the post partum period is described with emphasis on teaching self-help techniques such as: special nutritional considerations, how to recognize potential problems, and how to fit and adapt appliances to the changing body form of the patient as the pregnancy progresses. Phenomenologic information about the pregnant ostomate's perceptions about her pregnancy experience is also presented. Psycho-prophylaxis methods of complementary therapy alternatives alleviating nausea and vomiting within pregnancy, as well as the pain of childbirth are also presented. Although nursing/midwifery management of a pregnant ostomy patient is a challenge, those challenges are surmountable and outcomes resemble those found in the general population. PMID- 14744502 TI - Public, nurse and medical practitioner attitude and practice of natural medicine. AB - In the past 20 years, the popularity of natural medicine has increased, with nurses and medical practitioners being amongst several groups of clinicians expressing a widespread interest in natural therapies. It is argued that the degree of natural therapy use and the attitudes of these practitioners toward natural medicine may not only influence patient exposure to these therapies, but also have some bearing on the future use of natural medicine. Hence, in a critical review of recent studies, the utilisation of natural therapies and the attitudes of nurses, medical practitioners and the public toward natural medicine are explored. PMID- 14744503 TI - An aura of confusion: 'seeing auras-vital energy or human physiology?' Part 1 of a three part series. AB - The first of three papers that considers claims made for the perception or detection of vital energy. Many systems of Complementary and Alternative Medicine (CAM) assume the existence of a vital force that mediates therapeutic efficacy, for example chi or qi in Traditional Chinese medicine. Vital energy directly perceived or imaged that surrounds living organisms is frequently termed the aura. This paper aims to show how phenomena that arise as a consequence of the normal functioning of the human visual system can be inappropriately offered as support of claims for the direct perception of vital energy or the aura. Specifically, contrast and complementary colour phenomena, entoptic phenomena and the deformation phosphene, the 'flying corpuscle effect', the blind spot and the 'reverse telescope effect' are explained and discussed. PMID- 14744504 TI - Use of anti-emetic herbs in pregnancy: women's choices, and the question of safety and efficacy. AB - The majority of North American pregnant women experience some degree of nausea and vomiting, usually in the first few months of pregnancy. Women utilize many coping strategies, including self-treatment with herbal medicine and other alternative therapies. In a qualitative study of self-care in pregnancy, birth and lactation within a non-random sample of 27 women in British Columbia, Canada, 20 women (74%) experienced pregnancy-induced nausea. Ten of these women used anti emetic herbal remedies, which included ginger, peppermint, and Cannabis. The safety and efficacy of each of these herbal remedies is discussed here. Only ginger has been subjected to clinical trials among pregnant women, though all three herbs were clinically effective against nausea and vomiting in other contexts, such as chemotherapy-induced nausea and post-operative nausea. While safety concerns exist in the literature for all three herbs with regards to their use by pregnant women, clinical evidence of harm is lacking. PMID- 14744505 TI - A holistic model of advocacy: factors that influence its use. AB - Although advocacy is embraced by nursing as an essential component of holistic philosophy, its scope is often limited in practice. In this article, a research study that examined the use of an expanded definition of advocacy is described. A link to the role of advocacy as a complementary therapy and in relation to facilitating the use of complementary therapies by patients is provided. Fifty two registered nurses completed a researcher developed advocacy research instrument that assessed the use of moral-ethical, legal, political, spiritual, and substitutive advocacy along with various factors thought to influence the use of advocacy including moral development, perceived assertiveness, and perceived job security. An additional 40 RN-BSN students generated case studies of advocacy enacted in practice that were used as examples of the five categories of advocacy and to support the findings of the survey. Results indicated that moral-ethical advocacy was used more often than the other four categories. Moral stage development had a significant effect on substitutive advocacy but assertiveness and job security were not significant factors influencing any category of advocacy. PMID- 14744506 TI - Therapeutic Eurythmy-movement therapy for children with attention deficit hyperactivity disorder (ADHD): a pilot study. AB - This paper considers Therapeutic Eurythmy (TE) as a possible therapy for children with attention deficit hyperactivity disorder (ADHD).ADHD manifests as a complex psychological disturbance in which deficit of attention such as forgetfulness or distraction is the main symptom. It would seem that a growing number of children seem to be affected by this syndrome and an increasing number of alternative approaches to treatment are being sought. Therapeutic Eurythmy is a movement Therapy in the context of anthroposophical medicine. As a holistic therapy TE affects both physical and spiritual aspects of illness. TE entails the practice of movement exercises learned from a trained therapist. In this exploratory study, the effects of TE on behavioural functioning were examined.This paper addresses five single cases where the therapy induced positive changes to client's attention span, concentration, tempo of work and motor skills such as coordination, dexterity and social behaviour. Standard psychological tests parameters for movement, and for attention were used to assess aspects of performance.A positive shift was observed with reference to concentration and development of movement skills. Results were less pronounced but showed positive improvements on working speed and social behaviour problems. Hyperactivity also diminished to some extent. Generally, children were considered to be more mature in their development after therapy. The results of these case studies suggest that TE may be helpful for children with ADHD. However, more systematic research is warranted. PMID- 14744507 TI - Countering misleading information. AB - Orthodox medicine generally demands evidence in the form of randomised controlled trials (RCTs) before accepting the value of a particular therapy/intervention from the CAM field. Yet many RCTs are badly executed as they are carried out by doctors or scientists rather than holistic practitioners, and peer reviewers for conventional medical journals may not have sufficient knowledge to be able to assess a CAM paper properly. This article discusses inadequacies found in RCTs and other papers related to CAM, and pinpoints how research should be critically evaluated and reviewed. Examples are taken from the fields of aromatherapy, herbalism, acupuncture/TCM and homeopathy. The aim of this paper is to highlight common misunderstandings and misguided assumptions that may arise when undertaking research in the field of complementary medicine that may result in erroneous conclusions being drawn from data and which may have far reaching implications for clinical practice. The STRICTA recommendations for acupuncture are discussed. PMID- 14744508 TI - Critical incident: idiosyncratic allergic reactions to essential oils. AB - Essential oils have the potential to initiate allergic reactions due to their volatile and skin absorbent nature. Practitioners and aromatherapy teachers need to be aware of the potential for allergies and be equipped to deal with them if they should arise. Two cases are presented of potentially serious reactions that occurred within a learning situation along with a brief literature critique about allergic reactions to essential oils. PMID- 14744509 TI - Learning from dolphins. AB - Dolphins have been shown to have a powerful impact on the wellbeing of humans, how do they do it? This article reflects the thoughts of one person after spending time with these wonderful creatures. PMID- 14744519 TI - A comparison of flashbacks and ordinary autobiographical memories of trauma: content and language. AB - We investigated hypotheses derived from the dual representation theory of posttraumatic stress disorder, which proposes that flashbacks and ordinary memories of trauma are supported by different types of representation. Sixty-two participants meeting diagnostic criteria for posttraumatic stress disorder completed a detailed written trauma narrative, and afterwards identified those sections in the narrative that had been written in flashback and ordinary memory periods. As predicted, flashback periods were characterised by greater use of detail, particularly perceptual detail, by more mentions of death, more use of the present tense, and more mention of fear, helplessness, and horror. In contrast, ordinary memory sections were characterised by more mention of secondary emotions such as guilt and anger. PMID- 14744520 TI - The Speech Anxiety Thoughts Inventory: scale development and preliminary psychometric data. AB - Cognitions have been known to play a central role in the development, maintenance, and treatment of speech anxiety. However, few instruments are currently available to assess cognitive contents associated with speech anxiety. This report describes three studies examining the psychometric characteristics of a revised English version of the Speech Anxiety Thoughts Inventory (SATI)-an instrument measuring maladaptive cognitions associated with speech anxiety. In Study 1, factor analyses of the SATI revealed a two-factor solution-"prediction of poor performance" and "fear of negative evaluation by audience", respectively. In Study 2, the two-factor structure was replicated. In addition, results revealed stability over a four-week period, high internal consistency, and good convergent and discriminant validity. In Study 3, the scale demonstrated sensitivity to change following brief exposure-based treatments. These findings suggest that the SATI is a highly reliable, valid measure to assess cognitive features of speech anxiety. PMID- 14744521 TI - Correcting distorted perception of sleep in insomnia: a novel behavioural experiment? AB - Patients with primary insomnia overestimate their sleep onset latency (SOL) and underestimate their total sleep time (TST). The present study aimed to test the utility of a novel behavioural experiment designed to correct distorted perception of sleep among patients diagnosed with primary insomnia. Individuals with primary insomnia were asked to wear an actigraph and keep a sleep diary for three nights. On the following day, half were shown the discrepancy between the data recorded on the actigraph and their sleep diary (Shown-Discrepancy Group), the other half were not shown the discrepancy (No-Demonstration Group). Participants were then asked to wear the actigraph and keep a sleep diary for three further nights. Following the behavioural experiment, the Shown-Discrepancy Group estimated their SOL more accurately and reported less anxiety and preoccupation about sleep compared to the No-Demonstration Group. The theoretical and clinical implications of these findings are discussed. PMID- 14744522 TI - Beyond abuse: the association among parenting style, abdominal pain, and somatization in IBS patients. AB - This study assessed the relative strength of the association between abuse, negative parenting style, and somatization in irritable bowel syndrome (IBS) patients. Drawing from preclinical stress physiology and abuse research identifying the family social climate as a frequently stronger and independent determinant of long-term health effects than abuse-specific variables, we predicted that negative parenting behaviors would more strongly correlate with somatization than abuse. Subjects were 81 consecutively evaluated patients, who at baseline underwent psychological testing, measuring perceived parental style, abuse history, somatization, and pain. Although abuse correlated with maternal and paternal rejection, abuse was not associated with somatization. Higher levels of rejection and/or hostility among fathers (not mothers) were more strongly correlated with somatization than was abuse. Further, paternal parenting behaviors were more predictive of somatization than abuse, age, and gender. The lack of an association between abuse and somatization is discussed in light of limitations of biopsychosocial IBS models, whose strong focus on "pathological stressors" (e.g., abuse, trauma) as risk factors may overlook the importance of "less extreme" parenting variables in influencing somatic complaints. The relationship between parenting and somatization is discussed in the context of broader behavioral science research linking disruptions in the quality of parenting to dramatic and long-term changes in patterns of stress reactivity and brain abnormalities seen in IBS patients. PMID- 14744523 TI - Diagnostic efficiency of DSM-IV criteria for obsessive compulsive personality disorder in patients with binge eating disorder. AB - This study examined the diagnostic efficiency of the DSM-IV criteria for obsessive compulsive personality disorder (OCPD) in patients with binge eating disorder (BED). Two hundred and eleven consecutive adult patients with axis I diagnoses of BED were reliably assessed with semi-structured diagnostic interviews. Conditional probabilities-sensitivity, specificity, positive predictive power (PPP), and negative predictive power (NPP)-were calculated for each of the eight criteria for OCPD, using the 'best-estimate' OCPD diagnosis as the standard. The diagnostic efficiencies of the OCPD criteria were variable, with three criteria failing to have predictive value (PPP<0.50). The best inclusion criterion (highest PPP) was 'Perfectionism,' which was also the overall most predictive criterion. The findings suggest ordering of the DSM-IV criteria for OCPD based on performance and call into question the utility of some criteria. PMID- 14744524 TI - Subtyping female adolescent psychiatric inpatients with features of eating disorders along dietary restraint and negative affect dimensions. AB - Cluster-analytic studies of eating disorders in adult patients have yielded two subtypes (pure dietary and mixed dietary-negative affect). This study aimed to replicate the subtyping in female adolescent psychiatric inpatients with features of eating disorders. Cluster analyses of 137 patients with eating-disordered features revealed a dietary-negative affect subtype (43%) and a pure dietary subtype (57%). The dietary-negative affect subtype was characterized by greater likelihood of binge eating, greater eating-related psychopathology, and greater body image dissatisfaction. The two subtypes did not differ significantly in scores reflective of clinical syndromes (other than the significantly higher depressive affect in the negative affect subtype), but the dietary-negative affect subtype was characterized by greater personality disturbance and higher reported concerns in clinical areas, including suicidality and childhood abuse. The cluster analysis produced different results from an alternative approach to subtyping by vomiting. These findings provide further support for the reliability and validity of this subtyping scheme for eating pathology. Clinically, the findings suggest that the combination of dieting and negative affect signals a more disturbed variant of eating-disorder related psychopathology in female adolescent psychiatric inpatients. PMID- 14744525 TI - Factor structure and stability of the Anxiety Sensitivity Index in a longitudinal study of anxiety disorder patients. AB - The past decade witnessed considerable debate over the factor structure of the Anxiety Sensitivity Index (ASI), with an eventual consensus emerging that supported a hierarchically organized factor structure. The present study attempted to replicate and examine the overall stability and utility of the hierarchical ASI factor pattern using a large sample of outpatients participating in an ongoing longitudinal study of anxiety disorders. Results supported a hierarchical factor structure for the ASI consisting of three lower-order factors measuring physical concerns, mental incapacitation concerns, and social concerns, all of which loaded significantly on a single second-order factor. Correlational analyses show good test-retest reliability and consistent patterns of intercorrelation for these factor-derived subscales across a 10-month time frame. Additional analyses provide support for the discriminant validity of the ASI subscales with regard to individuals with specific anxiety disorders. The theoretical implications of these findings for future evaluations of anxiety sensitivity are discussed. PMID- 14744526 TI - Disgust domains in the prediction of contamination fear. AB - Previous research has shown a relationship between the emotion of disgust and the fear of contamination. Heightened sensitivity to disgust and increased concerns over contamination has been observed in various disorders, such as obsessive compulsive disorder (OCD) and specific phobias. However, there is a paucity of research identifying the specific domains of disgust that contribute to contamination fear. The present study soughts to determine which domains of disgust elicitors reliably predict scores on a measure of OCD contamination obsessions and washing compulsions. We further conducted exploratory analyses that examined differences in disgust sensitivity among individuals classified as high and low in contamination fear. Three hundred and twenty-three undergraduate participants completed two measures of disgust sensitivity (Disgust Scale; Disgust Emotion Scale) and a measure of contamination fear (Padua inventory, contamination obsessions and washing compulsions subscale). Stepwise multiple regression analyses indicated that contamination fear was best predicted by seven different disgust domains, thereby suggesting that contamination fear is accounted for by generalized, rather than domain-specific, disgust elicitors. The categories of disgust that predicted contamination fear appeared to have an underlying commonality of threat of contagion. The relationship between fear of contamination and disgust sensitivity was more pronounced for animal reminder disgust elicitors as opposed to core disgust elicitors. Results also showed that individuals classified as high in contamination fear scored significantly higher than the low contamination fear group on all disgust domains. Clinical and research implications regarding the interrelationships between fear, disgust, and the fear of contamination are discussed. PMID- 14744527 TI - Neuroticism and self-criticism associated with posttraumatic stress disorder in a nationally representative sample. AB - Broad and specific psychological traits may uniquely differentiate trauma victims with PTSD from trauma victims without PTSD, but there is a need for representative, population-based research. We investigated elevated neuroticism and self-criticism in association with the presence versus absence of PTSD in a nationally representative sample of adults who experienced a traumatic stressor. Respondents were from the National Comorbidity Survey Part II (N=5877) (). Individuals who experienced one or more traumatic events were selected (N=3238). In separate regression analyses, elevated levels of neuroticism and self criticism were each significantly associated with PTSD among men and women who had experienced one or more traumatic events. After controlling for types of traumas experienced and other previously identified factors (Bromet, Sonnega, & Kessler, 1998. American Journal of Epidemiology, 147, 353-361), neuroticism remained significantly associated with PTSD in women and both neuroticism and self-criticism remained significant in men. Evidence from this nationally representative sample of adults who experienced traumatic events suggests that self-criticism and especially the broad personality domain of neuroticism may represent robust psychological dimensions associated with the presence of PTSD. PMID- 14744528 TI - Comment on "Methylation patterns and K-ras mutations in tumor and paired serum of resected non-small-call lung cancer patients" (Cancer Letters 2003;193:207-216). PMID- 14744529 TI - Searching for cancer-associated gene polymorphisms: promises and obstacles. AB - Low-penetrance genetic variations appear to form the most essential component of the heritability of cancer risk. Search for relevant polymorphic candidates faces significant obstacles, due to both the high number of potentially promising single nucleotide polymorphisms (SNPs) and the intrinsic difficulties in identification of weak gene-disease interactions. At present, extensive case control studies can be applied only to a limited number of gene polymorphisms. Therefore, the choice of SNPs that deserve an exhaustive populational analysis is of primary importance. Preferences are usually given to those genetic pathways, whose variability and role in cancer causation have been already shown by prior studies. The available electronic databases and software tools may allow further SNP sorting, based on functional predictions. The design for the pilot study may need to be different from the one for large-scale case-control analysis. Some investigations justify non-random patient selection for preliminary assessment of low-penetrance effects, with the emphasis on particularly susceptible individuals (familial, early onset, multiple cancer cases). Other presumably accelerating approaches suggest a decisive exclusion of SNP candidates showing only marginal effects, relaxed formats for rapid dissemination of preliminary data, use of more demonstrative controls such as elderly tumor-free subjects, etc. These short-cuts cannot be properly validated for the time being, due to the paucity of identified low-penetrance risk modifiers. It is expected that the increasing capacities of available DNA collections, coupled with the rapid development of high-throughput genotyping technologies, will vastly accelerate the research on polygenic cancer susceptibility. PMID- 14744530 TI - Changes in cecal microbial metabolism of rats induced by individual and a mixture of drinking water disinfection by-products. AB - Disinfection of drinking water has been one of the greatest public health successes. Numerous halogenated disinfection by-products (DBPs) occur and chronic ingestion has been associated with an increased risk for colorectal cancer in human populations. Because the intestinal microbiota can bioactivate xenobiotics, studies have been performed to examine the effects of individual DBPs on intestinal microbial metabolism. No studies have been conducted on a defined mixture of DBPs to determine if there is an enhancement of response to a mixture. Ten-week-old male Long-Evans rats were treated in their drinking water for 17 weeks with 0.4 g/l potassium bromate, 1.8 g/l chloroform, 0.7 g/l bromodichloromethane (BDCM), 0.07 g/l 3-chloro-4-(dichloromethyl)-5-hydroxy-2(5H) furanone (MX), or a mixture of the four chemicals or distilled water. Cecal nitroreductase (NR), azoreductase (AR), dechlorinase (DC), beta-glucuronidase (GLR), beta-galactosidase (GAL), and beta-glucosidase (GLU) were assayed. No change in GLU or GLR activity was detected after treatment. BDCM treatment reduced DC and GAL activities and elevated NR and AR activity. GAL, AR, and NR activities were significantly different after treatment with bromate, chloroform, BDCM, and MX, but not the mixture. DC activity after chloroform-, MX-, or BDCM treatment was significantly below control levels. The present study shows that changes in intestinal microbial metabolism do occur after treatment with individual and a mixture of DBPs but the changes were not additive in the mixture group. PMID- 14744531 TI - Cellular accumulation of dietary anticarcinogenic isothiocyanates is followed by transporter-mediated export as dithiocarbamates. AB - Many dietary isothiocyanates (ITCs) are potent anticarcinogenic agents. ITCs rapidly accumulate to high concentrations in cells as a result of conjugation with intracellular thiols, especially glutathione (GSH). The anticarcinogenic activity of ITCs depends on, at least partly, their accumulation in cells. We report that three major anticarcinogenic ITCs, including allyl-ITC, benzyl-ITC, and phenethyl-ITC, were rapidly exported, upon accumulation in cells, mainly in the forms of GSH- and cysteinylglycine-conjugates, apparently involving MRP-1 and Pgp-1. These findings are consistent with our previous results regarding cellular export of another anticarcinogenic ITC, sulforaphane, and suggest a common cellular response to ITCs. PMID- 14744532 TI - DNA damage and repair efficiency in lymphocytes from schizophrenic patients. AB - In the present study we examined schizophrenic patients' lymphocytes sensitivity to the effects of external factors, such as hydrogen peroxide and gamma irradiation and also their repair efficiency with the comet assay. Our results did no show any difference in basal levels of DNA damage between schizophrenic and normal populations. The slightly increased sensitivity of the schizophrenic population to the externally induced DNA damage compared to controls was not statistically significant. Also the small reduction in the DNA repair efficiency in schizophrenics in comparison to normal population was found to be not statistically significant. Finally, patients with heritable predisposition to schizophrenia did not show any difference in their response from the other schizophrenics. PMID- 14744533 TI - Apoptosis-related gene expression after hyperthermia in human tongue squamous cell carcinoma cells harboring wild-type or mutated-type p53. AB - Hyperthermia is useful for the treatment of human head and neck cancer, as it is relatively easy to perform thermoregulation when compared with deep organs. In this study, we focused attention on the p53 as a predictive indicator of hyperthermic cancer therapy. We used two kinds of cell lines of a human squamous cell carcinoma (SAS) with identical backgrounds of function except for the p53 protein. We assayed the heat sensitivity, frequency of apoptosis, and apoptosis related gene expression after heat treatment using DNA array. The SAS/neo (wild type p53; wtp53) cells were sensitive to heat, and the induction of Caspase-3 activation and apoptosis in the wtp53 cells was clearly high compared with the SAS/mp53 (mutated p53; mp53) cells. The gene expression of apoptosis suppressive genes such as IL-12 p35 decreased in the wtp53 cells, and IL-12 R beta1 increased in the mp53 cells, though apoptosis-promotive genes of Caspase-9, CD30 and CD40 were induced p53-independently by hyperthermia. It is suggested that heat-induced apoptosis was suppressed by IL-12-related genes in the mp53 cells. These findings strongly imply that p53 status is a useful candidate for a predictive indicator of the effectiveness in hyperthermic therapy. PMID- 14744534 TI - Sensitivity of proteasome to its inhibitors increases during cAMP-induced differentiation of neuroblastoma cells in culture and causes decreased viability. AB - Inhibition of proteasome activity is associated with a reduction in proliferation and apoptosis in cancer cells, depending upon the extent of inhibition. We have reported that a minimal inhibition of proteasome activity prevented adenosine 3'5'-cyclic monophosphate (cAMP)-induced differentiation and caused apoptosis in murine neuroblastoma (NB) cells in culture. In order to establish whether an elevated cAMP level increases the sensitivity of proteasome to its inhibitors, MG 132 and lactacystin (proteasome inhibitors) were added concomitantly with a stimulator of adenylate cyclase (prostaglandin A1) and an inhibitor of cyclic nucleotide phosphodiesterase (RO20-1724). Results showed that concentrations of MG-132 that did not reduce or that minimally inhibited proteasome activity also did not affect the proliferation of undifferentiated NB cells. However, these concentrations of MG-132 in the presence of an elevated cAMP level markedly inhibited proteasome activity and caused extensive cell death. Similar results were obtained with lactacystin. In normal murine fibroblasts, cAMP-induced reduction in proliferation was not affected by any concentration of MG-132 used in this study. These results suggest that proteasome exhibits increased sensitivity to its inhibitors following an elevation of cAMP level in NB cells, but not in normal fibroblasts, and that this may account for the enhanced cell death in NB cells. Thus, the strategy of using low doses of a proteasome inhibitor in combination with a cAMP-stimulating agent may be useful in pre clinical and clinical studies on NB tumor because of the selectivity of the effect on cancer cells. PMID- 14744535 TI - Expression of the chondromodulin-I gene in chondrosarcomas. AB - We investigated the expression of the Chondromodulin-I (ChM-I) gene, a putative tumor suppressor gene in cartilaginous tumors, by quantitative RT-PCR in 15 chondrosarcomas (CSs). Eight CSs expressed the ChM-I gene at the level higher than those in articular cartilage (positive cases), whereas the expression of the ChM-I gene in the remaining seven CSs was lower than those in articular cartilage (negative cases). All of five peripheral CS were positive, and the ChM-I positive tumors shared expression profiles of cartilage-related genes with articular cartilage cells. On the other hand, all of four central CSs without extramedullary lesions were negative, and the ChM-I negative tumors expressed the parathyroid hormone-related peptide gene at the lower level and the COL10A1 genes at the higher level than articular cartilage cells. Neither the histological grade nor the rate of recurrence showed clear association with the level of ChM-I gene expression. These results suggested that the expression of ChM-I gene in CS has no direct role in tumorigenesis but rather reflects the site of tumor development and therefore precursor of tumor cells. PMID- 14744536 TI - ITIH5, a novel member of the inter-alpha-trypsin inhibitor heavy chain family is downregulated in breast cancer. AB - The inter-alpha-trypsin inhibitor (ITI) family constitutes a group of proteins built up from one light chain and a variable set of heavy chains. Originally identified as plasma protease inhibitors, recent data indicate that ITI plays a role in extracellular matrix (ECM) stabilization and in prevention of tumor metastasis. Here we describe cloning as well as phylogenetic and expression analysis of a novel member of the heavy chain gene family, ITIH5. ITIH5 contains the two domains conserved in all known ITIHs, the vault protein inter-alpha trypsin (VIT) domain and a von Willebrand type A (vWA) domain. However, ITIH5 diverged early from a common ancestor of the other subfamilies. We found strong downregulation of ITIH5 expression in breast tumors by real-time PCR and RNA in situ hybridization. While normal breast epithelial cells clearly express ITIH5, expression is consistantly lost or strongly downregulated in invasive ductal carcinoma. ITIH5 mRNA was neither detectable in cancerous nor benign breast cell lines. We propose that loss of ITIH5 expression may be involved in breast cancer development. PMID- 14744537 TI - Proteomic evaluation of core biopsy specimens from breast lesions. AB - Analysis of tumour samples by a proteomic technology, which combines two dimensional gel electrophoresis and mass spectrometry analysis, is a promising approach for molecular characterization of cancer. Proteomic analysis of neoplasms is usually performed on surgical material. The possibility to perform proteomic analysis on pre-operative samples might be useful for diagnostic purposes or for determination of tumour sensitivity to therapy. In this study, we report how tissues from core biopsy of breast lesions can be routinely used to obtain accurate protein expression profiles by proteomic analysis. Protein profiles from fibroadenomas were compared to those from ductal infiltrating carcinomas. By using mass spectrometry, identification of proteins overexpressed in carcinomas with respect to fibroadenomas was obtained. Thus, our study provides a methodology to perform proteomic analysis on pre-operative samples of breast lesions. PMID- 14744538 TI - Correlation between serum vascular endothelial growth factor and endostatin levels in patients with breast cancer. AB - Serum vascular endothelial growth factor (VEGF) and endostatin levels were detected in 59 patients with breast cancer before surgery and at 3 weeks after surgery. Pre-operatively, their levels were significantly elevated and correlated with each other. Post-operatively, VEGF level decreased significantly and endostatin remained at a high level. Patients with both normalized VEGF and elevated endostatin following surgery had a lower risk of relapse than patients whose VEGF failed to normalize. Univariate and multivariate analyses showed a correlation between elevated VEGF level and short free-relapse survival. These findings suggest a new angiogenesis balance is formed in the patients after surgery and such a resultant balance may be beneficial for the prognosis of breast cancer, which deserves more extensive study. PMID- 14744539 TI - Tumor dihydropyrimidine dehydrogenase in stage II and III colorectal cancer: low level expression is a beneficial marker in oral-adjuvant chemotherapy, but is also a predictor for poor prognosis in patients treated with curative surgery alone. AB - The aim of this study was to evaluate the prognostic significance of tumor dihydropyrimidine dehydroganase (DPD) in curatively resected colorectal cancer patients who received or did not receive oral 5-FU based-adjuvant chemotherapy. Among 182 patients with stage II-III colorectal cancers, 89 patients (adjuvant chemotherapy group) received oral 5-FU based-adjuvant chemotherapy, and 93 patients (surgery alone group) did not receive 5-FU. DPD expressions in the tumors and in the normal colonic mucosa were measured by enzyme-linked immunosorbent assays. The mean DPD expression of the tumors was significantly lower than that of the normal mucosa (54.4 +/- 40.4 versus 72.3 +/- 23.3 Unit/mg protein, P < 0.01). For survival analyses, we designated the cut-off value of tumor DPD as its median value (46.3). In the adjuvant chemotherapy group, high tumor DPD levels were associated with poor survival (HR, 5.24; P = 0.03). In the surgery alone group, high tumor DPD levels were associated with better survival (HR, 0.32; P = 0.02). In conclusion, tumor DPD level is an efficacious marker in oral 5-FU based-adjuvant chemotherapy for colorectal cancer; however, low tumor DPD predicts reduced survival in patients treated with curative surgery alone. PMID- 14744540 TI - Establishment and characterization of a biphasic synovial sarcoma cell line, SYO 1. AB - We describe here the establishment of a new synovial sarcoma cell line, SYO-1, derived from a biphasic synovial sarcoma that developed in the groin of a 19-year old female. The cell line was maintained for more than 70 passages (more than 24 months) in vitro. The SYO-1 cells in monolayer culture exhibited a spindle shape without conspicuous pleomorphism. Immunohistochemically, the cells were positive for vimentin, type IV collagen, S-100, mdm2, bcl-2, c-Met and c-Kit. Tumors developed by their implantation in nude mice histologically showed biphasic features that were composed of areas of fascicles of spindle cells and areas of compact proliferation of polygonal to ovoid cells, which occasionally formed epithelial plaque and expressed cytokeratin and EMA. SYO-1 cells harbored the characteristic t(X;18)(p11.2;q11.2) translocation by chromosome analysis and SYT SSX2 chimeric transcript by RT-PCR. The SYO-1 cells, the first characterized cell line derived from biphasic synovial sarcoma retaining the characteristic genetic and phenotypic features of the tumor, will be useful for various investigations on synovial sarcoma, especially for its epithelial differentiation. PMID- 14744541 TI - Components of early maternal environment affecting the predisposition to catalepsy. AB - Reciprocal pup substitution (cross-fostering) in cataleptic GC (designated so by the initials of words "genetic" and "catalepsy") and control Wistar females resulted in attenuation of cataleptic predisposition in GC rats fostered by Wistar foster-mothers. The latter demonstrate a more intense maternal care than GC females. There was a significant negative correlation between the frequency of mother staying in nest and the duration of pinch-induced catalepsy in pups fostered by her. In the home-cage retrieval test, the females of the strains compared showed a significant dependence of the latencies of approach to, and retrieval of, pups on their own and the pups' genotype. PMID- 14744542 TI - Mild mutations in the pan neural gene prospero affect male-specific behaviour in Drosophila melanogaster. AB - The fruitfly Drosophila melanogaster is one of the most appropriate model organisms to study the genetics of behaviour. Here, we focus on prospero (pros), a key gene for the development of the nervous system which specifies multiple aspects from the early formation of the embryonic central nervous system to the formation of larval and adult sensory organs. We studied the effects on locomotion, courtship and mating behaviour of three mild pros mutations. These newly isolated pros mutations were induced after the incomplete excision of a transposable genomic element that, before excision, caused a lethal phenotype during larval development. Strikingly, these mutant strains, but not the strains with a clean excision, produced a high frequency of heterozygous flies, after more than 50 generations in the lab. We investigated the factors that could decrease the fitness of homozygotes relatively to heterozygous pros mutant flies. Flies of both genotypes had slightly different levels of fertility. More strikingly, homozygous mutant males had a lower sexual activity than heterozygous males and failed to mate in a competitive situation. No similar effect was detected in mutant females. These findings suggest that mild mutations in pros did not alter vital functions during development but drastically changed adult male behaviour and reproductive fitness. PMID- 14744543 TI - Novelty-seeking behavior and stress-induced locomotion in rats of juvenile period differentially related to morphine place conditioning in their adulthood. AB - The relationship between motor responses in a novel environment and susceptibility to place conditioning effect of psychostimulants has been reported in adult rats. However, it is in question whether this correlation could be generalized to motor activity in rats of juvenile period and place conditioning effect in their adulthood for narcotic morphine. In the present study, we tested locomotor activity in an arena open-field and the subsequent novelty-seeking behavior after adaptation process in juvenile rats (P42) and morphine (2 mg/kg) place conditioning effect 56 days later in the same rats' adulthood (P98). Our results showed that rats with high response to novelty (HRN) spent more prolonged duration in the drug-paired compartment in the place conditioning test compared with their low response counterparts (LRN), with the latter group no salient change on this measure. Moreover, rats with high response to the open-field test (HRS) expressed equally elevated duration in drug-paired side relative to their low response counterparts (LRS). The present research demonstrated that novelty seeking behavior and locomotor activity in the open-field in rats of juvenile period differentially related to morphine place conditioning in their adulthood, with slow acquisition of morphine place conditioning effect in LRN animals. PMID- 14744544 TI - Pigeon's recognition of cartoons: effects of fragmentation, scrambling, and deletion of elements. AB - J. Cerella [Pattern Recognit. 12 (1980) 1] and more recently S. Watanabe [Behav. Proc. 53 (2001) 3] demonstrated that pigeons showed no decrement in recognizing cartoons that were spatially scrambled, indicating that pigeons' discriminative responding is controlled by local features alone. In contrast Kirkpatrick-Steger et al. [J. Exp. Psychol. Anim. Behav. Proc. 24 (1998) 34] used line drawings as stimuli and demonstrated the importance of spatial organization for picture recognition by pigeons, confirming related findings reported in their previous studies. The present study revisited the recognition of cartoons by pigeons. In Experiment 1, pigeons were trained to discriminate cartoon people on a variety of background scenes. Subsequent tests revealed that discriminative performances with both familiar and novel instances decreased as the objects and object-like parts were progressively fragmented, indicating that search for the targeted cartoon people in the stimulus array might have enhanced the pigeons to attend to global aspects of cartoon people. Experiment 2 used line drawings of cartoon faces as stimuli and examined effects of scrambling and deletion of components. A set of components (eyes and eyebrows) exerted strong control over behavior and scrambling only moderately suppressed responding. The results suggest that pigeons use both global and local aspects, with different mixtures of these types of information depending on the particular perceptual context. PMID- 14744545 TI - Delay discounting and probability discounting as related to cigarette smoking status in adults. AB - This study examined relations between adult smokers and non-smokers and the devaluation of monetary rewards as a function of delay (delay discounting, DD) or probability (probability discounting, PD). The extent to which individuals discount value, either as a function of a reward being delayed or probabilistic, has been taken to reflect individual differences in impulsivity. Those who discount most are considered most impulsive. Previous research has shown that adult smokers discount the value of delayed rewards more than adult non-smokers. However, in the one published study that examined probability discounting in adult smokers and non-smokers, the smokers did not discount the value of probabilistic rewards more than the non-smoker controls. From this past research, it was hypothesized that measures of delay discounting would differentiate between smokers and non-smokers but that probability discounting would not. Participants were 54 (25 female) adult smokers (n = 25) and non-smokers (n = 29). The smokers all reported smoking at least 20 cigarettes per day, and the non smokers reported having never smoked. The results indicated that the smokers discounted significantly more than the non-smokers by both delay and probability. Unlike past findings, these results suggest that both delay and probability discounting are related to adult cigarette smoking; however, it also was determined that DD was a significantly stronger predictor of smoking than PD. PMID- 14744546 TI - The effects of concurrent task and gap events on peak time in the peak procedure. AB - The effect of a concurrent task on timing performance of pigeons was investigated with the peak interval procedure. Birds were trained to peck a side key on a discrete-trial schedule that included reinforced fixed-interval (FI) 30-s trials and nonreinforced extended probe trials. Then, in separate sessions, birds were trained to peck a 6-s center key for food. In a subsequent test phase, the FI procedure was in effect along with dual-task probe test trials. On those test trials, the 6-s center key (task cue) was presented at 3, 9, or 15s after probe trial onset. During another test phase, a 6-s gap (the FI keylight was extinguished) was presented at 3, 9, or 15s after probe trial onset. Peak time increased with center key time of onset, and was greater under task than gap conditions. Moreover, peak time under task conditions exceeded values predicted by stop and reset clock mechanisms. These results are at variance with current attentional accounts of timing behavior in dual-task conditions, and suggest a role of nontemporal factors in the control of timing behavior. PMID- 14744548 TI - Effect of behavioral history on subsequent responding: experiments using rotary fixed-ratio schedules. AB - The present study proposes a novel experimental approach to examine the effect of previous schedule history on subsequent responses. College students used a three button device to indicate their choice of response. We performed two experiments. Each experiment consisted of three phases. In experiment I, fixed-ratio 10 (FR 10) reinforcement schedule was employed in phases 1 and 3. Phase 2 employed "Rotary-FR 10" schedule: after obtaining a point produced by pressing one of three specified buttons in a triangular array 10 times, the effective button changed to another in a clockwise direction. Optimal response pattern for Rotary FR 10 was analyzed. Two types of behavior, referred to as "consecutive" and "scatter" were observed in phase 1. The mean number of optimal responses in consecutive type was significantly higher than scatter type in phases 2 and 3. In experiment II, two schedules were used in phase 1, and the same protocol used in experiment I was followed. We found that the reinforcement schedule used in phase 1 affected acquisition of the optimal response in phase 2. These results suggest that the earlier behavioral tendencies play an important role in determining subsequent behavior and that such behavioral tendency can be modified by specifically designed protocols. PMID- 14744547 TI - Need probability effects in animal short-term memory. AB - Five pigeons performed in a delayed matching-to-sample (DMTS) procedure with five delay durations (0.5, 2.5, 5, 10 and 20 s) mixed within sessions. Contrary to the predictions of need probability theory, discriminability decreased when fewer short than long delays were included in each session. To test whether the decrease in discriminability was due to a decrease in obtained reinforcement at short delays, the number of trials at each delay was held constant and reinforcer probability was increased with increasing delay. This manipulation produced a similar decrease in discriminability as when the frequency of delays was manipulated. It was concluded that the effect of delay frequency on the forgetting function is mediated by the effect of the reinforcer distribution, which influences discriminability by weakening stimulus control. PMID- 14744549 TI - Comparison of social ranks based on worm-running and aggressive behaviour in young domestic fowl. AB - Worm-running is behaviour in which a chick runs carrying a worm-like object while flock mates follow and attempt to grab the object from its beak. We hypothesised that social ranks based on worm-running frequency are stable over time and are positively correlated with social ranks based on success in aggressive interactions when older. At 8-12 days of age, we scored worm-running in 17 groups of 12 female White Leghorn chicks during three 10-min tests. Based on instantaneous scans at 5-s intervals, the bird carrying the 'worm' most often was placed in rank one and so on down the rank order. These tests were repeated at 68 70 days of age. An aggression index for each bird was calculated as the number of aggressive acts given, divided by the number given and received, during three 1-h observation periods when the birds were 68-70 days. Ranks obtained in worm running tests were positively correlated over the two age periods (P < 0.05) but were not correlated with ranks based on the aggression index (P > 0.05). Our results indicate that worm-running ranks are not predictive of success in aggressive interactions. Instead, worm-running fits some criteria for play. PMID- 14744550 TI - Tool-use and tool-making by captive, group-living orangutans (Pongo pygmaeus abelii) at an artificial termite mound. AB - The present study examined the use and making of tools to obtain foodstuffs in artificial-mound holes by five captive, group-living Sumatran orangutans (Pongo pygmaeus abelii). Three adult orangutans frequently stripped leaves and twigs from a branch provided (tool-making), and then inserted the tool into a hole to obtain foodstuffs (tool-using). A 5-year-old female juvenile usually used the tools that adult orangutans had previously used, but rarely made tools herself. A 2-year-old male infant did not use any tools. The adult orangutans tend to leave one to several leaves at the top of the branch than to leave many leaves on the branch or to strip all leaves. It seemed likely that tools with appropriate leaves are easier to insert into holes and obtain more foodstuffs, compared with branches with many leaves or sticks without any leaves. When the orangutans were unable to insert a tool into a hole, they usually modified the tool and/or changed their tool-using technique, such as changing how they grasped the tool. These findings are discussed from the perspectives of the orangutan's behavioral flexibility regarding tool-use skills and hierarchical organization in food processing techniques. PMID- 14744551 TI - Adaptation level effects in discrimination of flicker frequency. AB - Pigeons accustomed to food reinforcement for responding in the presence of a 25 Hz flickering light were exposed to several sets of flicker-frequency stimuli arranged as increasing and decreasing series. In the first experiment, food was occasionally delivered for key pecks during 30-s periods of 25-Hz flicker appearing at the beginning, midway, and at the end of an ascending and descending series of nine frequencies, ranging from 13 to 37 Hz. These stimuli appeared for 15-s periods with no food available (extinction). Gradients of responding to flicker values in the ascending series differed from those in the descending series, showing displacements in peak responding toward the lower and higher frequency values, respectively. The same effects occurred when the sequence was changed so that a descending series was followed by an ascending series of frequencies. These effects are consonant with an adaptation level (AL) interpretation and were replicated in a second experiment in which durations of the extinction presentations were increased to 30s. In a final condition, only a descending series was presented and displacement of peak responding from 25 Hz to a higher frequency stimulus, 28 Hz, was observed. PMID- 14744552 TI - Malaria: integrated approaches for prevention and treatment. PMID- 14744553 TI - Trends in malaria cases in Japan. AB - Just after World War II, more than 10,000 malaria cases per year were reported in Japan, including indigenous, imported and induced malaria. Malaria has been successfully eradicated since 1961 in Japan and now only imported malaria cases are encountered. However, as the number of Japanese people who are going abroad and also the number of foreigners who are visiting Japan increases (about 16 and 5 millions, respectively, in 2001), so does the chance for Japanese doctors to see imported malaria or transfusion-transmitted malaria cases. In fact, the total number of the patients with acute malaria in Japan has been around 100-150 annually for the last 10 years. Of those, about 75% are Japanese and 25% are foreigners, and about 75% are male and 25% are female. The peak age is in the 20s. Recently, about 45% of patients are Plasmodium falciparum and another 45% Plasmodium vivax infections. The former species is likely to be seen in travelers coming back from African countries and the latter is mainly from Asian countries. The important issue is that patients in Japan have not been diagnosed promptly nor treated properly because doctors in Japan are no longer familiar with tropical medicine. Therefore, some patients are dying from severe malaria as a consequence. As it is, most of the effective medicines for drug-resistant malaria or severe malaria have not been registered in Japan. There is now a need for medical practitioners to focus on travel medicine in Japan. PMID- 14744554 TI - A potential role of interleukin 18 in severe falciparum malaria. AB - Interleukin-18 (IL-18) is a potent proinflammatory cytokine that induces interferon-gamma (IFN-gamma) production from Th1 cells, NK cells and activated macrophages, particularly in the presence of IL-12. However, it is also shown that without help from IL-12, IL-18 is capable of inducing IL-4 and IL-13 production in T cells, NK cells, mast cells and basophils, and that administration of IL-18 in conjunction with an allergen increases serum IgE levels. In order to clarify the role of IL-18 in disease severity of falciparum malaria, we have examined serum levels of IL-18, IFN-gamma, and IgE for 96 patients with falciparum malaria [Trans. R. Soc. Trop. Med. Hyg. 97, 236-241]. Results suggested that IL-18 plays a key role in inducing severe malaria through a pathway of elevating IFN-gamma, rather than its IgE inducing activity. Based on these results, the role of IL-18 in severe falciparum malaria will be discussed in this review. PMID- 14744555 TI - Channels and transporters as drug targets in the Plasmodium-infected erythrocyte. AB - Throughout the intraerythrocytic phase of its lifecycle the malaria parasite is separated from the extracellular medium by the plasma membrane of its host erythrocyte and by the parasitophorous vacuole in which the parasite is enclosed. The intracellular parasite itself has, at its surface, a plasma membrane, and has a variety of membrane-bound organelles which carry out a range of biochemical functions. Each of the various membranes of the infected cell have in them proteins that facilitate the movement of molecules and ions from one side of the membrane to the other. These 'channels' and 'transporters' play a central role in the physiology of the parasitised cell. From a clinical viewpoint they are of interest both as potential targets in their own right, and as potential drug targeting routes capable of mediating the entry of cytotoxic drugs into the appropriate compartment of the infected cell. In this review both of these aspects are considered. PMID- 14744556 TI - The effectiveness of impregnated bed net in malaria control in Laos. AB - Impregnated bed net (IBN) were used in 366 villages in the central and southern three provinces of Lao PDR from 1999 to 2000. It was confirmed that 81.0% of 40000 bed nets, which were donated by Japanese Grant Aid, were delivered within 2 years. The strengthening of information network systems in anti-malaria and strong relationship between community and local authorities ensured the success of operation in a short period. The number of patients and the slide positive rate of malaria decreased markedly in public health facilities in three provinces after the use of IBN. An entomological survey was conducted in Boualapha district, where malaria is endemic, to investigate the IBN efficacy on malaria vector. The density and parous rate of Anopeles dirus, which is the main malaria vector in the area, were markedly decreased in the village where IBN was used. This mosquito's behavior, which was baiting mainly humans during the time when the inhabitants sleep in the IBN, was considered to be advantageous in preventing malaria infection using by IBN. The area of distribution of A. dirus is similar to the high endemic area of malaria in Lao PDR. Thus, it is expected that the expansion of the IBN program in the southern provinces will lead to successful malaria control in subsequent years. PMID- 14744557 TI - Reduced microcirculatory flow in severe falciparum malaria: pathophysiology and electron-microscopic pathology. AB - The pathophysiology of severe falciparum malaria is complex, but evidence is mounting that its central feature is the old concept of a mechanical microcirculatory obstruction. Autopsy studies, but also in vivo observations of the microcirculation, demonstrate variable obstruction of the microcirculation in severe malaria. The principal cause of this is cytoadherence to the vascular endothelium of erythrocytes containing the mature forms of the parasite, leading to sequestration and obstruction of small vessels. Besides, parasitized red cells become rigid, compromising their flow through capillaries whose lumen has been reduced by sequestered erythrocytes. Adhesive forces between infected red cells (auto-agglutination), between infected and uninfected red cells (rosetting) and between uninfected erythrocytes (aggregation) could further slow down microcirculatory flow. A more recent finding is that uninfected erythrocytes also become rigid in severe malaria. Reduction in the overall red cell deformability has a strong predictive value for a fatal outcome. Rigidity may be caused by oxidative damage to the red blood cell membrane by malaria pigment released at the moment of schizont rupture. Anti-oxidants, such as N-acetylcysteine can reverse this effect and are promising as adjunctive treatment in severe malaria. PMID- 14744558 TI - Reduced levels of transforming growth factor-beta1, interleukin-12 and increased migration inhibitory factor are associated with severe malaria. AB - In the present study, we investigated plasma levels of interleukin (IL)-12 and transforming growth factor (TGF-beta1) in malaria patients as these two cytokines regulate the balance between pro- and anti-inflammatory cytokines. We compared plasma IL-12 and TGF-beta1 levels in groups of malaria patients categorized as uncomplicated, severe, cerebral and placental malaria. Both TGF-beta1 and IL-12 levels were significantly reduced in peripheral plasma of adults with severe and cerebral malaria as well as in plasma of Tanzanian children with cerebral malaria (P<0.05). Similar results were observed with both placental and peripheral plasma of pregnant women who were infected with Plasmodium falciparum. IL-18, a cytokine known to be critical for the induction of IFN-gamma along with IL-1, was produced more in uncomplicated adult patients than in aparasitimic healthy controls (P<0.05). However, IL-18 response rate declined as the symptoms of the disease became more severe suggesting that the IL-18 response may be impaired with increased malaria severity. Together, the results of the three cytokines support the notion that imbalance between pro- and anti-inflammatory cytokines may contribute to the development of severe malaria infection. With malaria infection during pregnancy, we demonstrated that macrophage migration inhibitory factor (MIF) levels in infected placental plasma were significantly higher than those in the paired peripheral plasma (P<0.05). MIF, therefore, may play an important role in the local immune response to placental P. falciparum infection. PMID- 14744559 TI - Development of artemisinin and its structurally simplified trioxane derivatives as antimalarial drugs. AB - Artemisinin and simplified trioxane analogs constitute a promising class of antimalarial chemotherapeutic agents. Their development since the early 1970s into clinical trials and clinical use has drawn much attention from medical scientists worldwide although the crude extract containing artemisinin has been used in China for treatment of fever for many centuries. Many research groups have independently and collaboratively conducted various studies on the artemisinin system both in search for the new compounds more antimalarially active than the parent artemisinin and in an attempt to understand its molecular mechanism(s) of action. Ongoing studies have provided a better understanding of the putative intermediates essential for the antimalarial activity and have led to designer trioxanes whose chemical structures have been simplified and modified to increase efficacy while lowering toxicity. Other desirable features beneficial to clinical uses such as bioavailability, drug stability and water solubility have been considered, and portions of the trioxane skeleton have been added or modified to accommodate these parameters accordingly. PMID- 14744560 TI - The pharmacodynamic study of a potent new antimalarial (MC1). AB - 2,3-bis(Trifluoromethyl)-4-(3-hydroxyquinuclidinylquinoline) or MC(1) is a new synthetic compound with potent antimalarial activity in vitro and in vivo studies. The IC(50) values of MC(1) and chloroquine in in vitro culture of Plasmodium falciparum are 7.0x10(-8) and 6.06x10(-7)M, respectively. In an in vivo study using Plasmodium berghei infected mice as the test model, the survival time of the infected mice without drug treatment was 6.00+0.58 days. Chloroquine and MC(1) at an equal dose of 7.5mg/kg, orally administered once daily for 4 days, prolonged the survival time of the infected mice from 6 to 14 days, and more than 28 days, respectively. At the doses that exhibit potent antimalarial activity in vivo, there are no observable toxic effects. Preliminary studies of the pharmacodynamic activity of this newly synthesized compound revealed that at the doses which exhibit potent antimalarial activity, there is no alteration in motor activity such as distance traveled, rotational behavior, and stereotypic activity. The blood glucose was not significantly altered. In the spontaneous beating, isolated right atria of mice, MC(1) exhibits direct negative chronotropism at high concentrations (10(-4)M). This effect is augmented in hyper K(+) bathing solution. A direct negative chronotropic effect was also observed when mefloquine at 5x10(-5)M was used. Preliminary pharmacodynamic study suggested that MC(1) is a potential new antimalarial drug that should be studied further. PMID- 14744561 TI - Therapeutic responses to antimalarial and antibacterial drugs in vivax malaria. AB - Plasmodium vivax is the most prevalent malaria infection and is an important cause of morbidity in Central and South America and Asia. P. vivax is generally sensitive to the common antimalarial drugs but high level resistance to chloroquine and/or pyrimethamine has been documented in some geographic locations. In the studies reviewed here, the therapeutic responses to antimalarial and antibacterial drugs in vivax malaria have been assessed in the Bangkok Hospital for Tropical Diseases. The evaluated drugs consisted of the eight most widely used antimalarial drugs and anti-bacterial drugs that possess antimalarial activities (tetracycline, doxycycline, clindamycin or azithromycin). The activities of these drugs in descending order of parasite clearance times were artesunate, artemether, chloroquine, mefloquine, quinine, halofantrine, primaquine, followed by the antibacterial drugs and lastly sulfadoxine pyrimethamine. Clinical responses to sulfadoxine-pyrimethamine were also poor with evidence of high grade resistance in 42% of the patients. Of the four antibacterial drugs, clindamycin was more effective than azithromycin and can be an alternative to the tetracyclines. Except for chloroquine and mefloquine which have long plasma half lives and may therefore suppress first relapses, the cumulative cure rates for the short acting antimalarial drugs were similar. Double infection with Plasmodium falciparum was common and usually manifested 3-4 weeks following clearance of vivax malaria. The prevalence of cryptic falciparum malaria was 8-15% and was higher in patients treated with less potent antimalarial drugs. Follow-up studies have revealed that the relapse time in Thai patients with vivax malaria is on average only 3 weeks, but can be suppressed by the slowly eliminated antimalarial drugs such as chloroquine and mefloquine. For accurate comparison of relapse/recrudescence rates in vivax malaria, at least 2 month's follow-up is required. It can be concluded that in malarious areas of Thailand, double infection with P. falciparum and P. vivax is common affecting at least 25% of the patients and usually manifests as sequential illnesses. P. vivax in Thailand is sensitive to chloroquine but has acquired high grade resistance to sulfadoxine-pyrimethamine. PMID- 14744563 TI - The hexose transporter of Plasmodium falciparum is a worthy drug target. AB - Despite substantial efforts at control over several decades, malaria is still a major global health problem as chemotherapy of malaria parasites is limited by established drug resistance and lack of novel treatment options. Intraerythrocytic stages of these parasites are wholly dependent upon host glucose for energy and malarial proteins involved in hexose permeation are therefore attractive new drug targets. For Plasmodium falciparum, the causative agent of severe malaria, a facilitative hexose transporter (PfHT), encoded by a single-copy gene mediates glucose uptake. We first established heterologous expression in Xenopus laevis to allow functional characterisation of PfHT. This review describes the value of using Xenopus oocytes in heterologous studies of P. falciparum-encoded proteins and summarises the properties of PfHT. Comparisons between Gluts (mammalian facilitative hexose transporters) and PfHT using this expression system have highlighted important mechanistic and structural differences between parasite and host proteins. Certain O-methyl derivatives of glucose proved particularly useful discriminators between mammalian transporters and PfHT. We exploited this selectivity and synthesised a long-chain O-3-hexose derivative (compound 3361) that potently inhibits PfHT expressed in oocytes and also kills P. falciparum when it is cultured in medium containing either glucose or fructose as a carbon source. To extend our observations to the second most important human malarial pathogen, we have cloned and expressed the Plasmodium vivax orthologue of PfHT, and demonstrate inhibition of glucose uptake by compound 3361. These findings validate malarial hexose transporters as a novel target. We now aim to design a new class of antimalarials by the discovery of highly specific inhibitors which could act with a broad spectrum of action on different Plasmodium spp. infections. PMID- 14744562 TI - Microfluidic approaches to malaria detection. AB - Microfluidic systems are under development to address a variety of medical problems. Key advantages of micrototal analysis systems based on microfluidic technology are the promise of small size and the integration of sample handling and measurement functions within a single, automated device having low mass production costs. Here, we review the spectrum of methods currently used to detect malaria, consider their advantages and disadvantages, and discuss their adaptability towards integration into small, automated micro total analysis systems. Molecular amplification methods emerge as leading candidates for chip based systems because they offer extremely high sensitivity, the ability to recognize malaria species and strain, and they will be adaptable to the detection of new genotypic signatures that will emerge from current genomic-based research of the disease. Current approaches to the development of chip-based molecular amplification are considered with special emphasis on flow-through PCR, and we present for the first time the method of malaria specimen preparation by dielectrophoretic field-flow-fractionation. Although many challenges must be addressed to realize a micrototal analysis system for malaria diagnosis, it is concluded that the potential benefits of the approach are well worth pursuing. PMID- 14744564 TI - Naphthoquine phosphate and its combination with artemisinine. AB - Naphthoquine phosphate and artemisinine are two antimalarials developed in China. Both drugs have proven to be efficacious and well tolerated as monotherapy as well as in combination in patients suffering from malaria. The Co-naphthoquine, a novel antimalarial combination, is an oral fixed combination tablet of the naphthoquine phosphate and artemisinine. Artemisinin is characterised by a rapid onset of schizonticidal action and a short half-life. Parasite clearance is, however, often incomplete when it is employed as a single agent unless high dosages are used over several days, but such a regimen may reduce patient compliance and increase the danger of toxicity. Naphthoquine phosphate, by contrast, has a slower onset of action and a longer half-life, associated with a low recrudescence rate. The two components act synergistically in animal, and clinically provide more rapid relief of symptoms and a higher cure rate than either component alone. The combination tablet was initially developed by the Academy of Military Medical Sciences (AMMS), Beijing, China. PMID- 14744565 TI - The physiological basis of transcranial motor cortex stimulation in conscious humans. AB - Transcranial stimulation of the human motor cortex can evoke several different kinds of descending activity depending on the type of stimulation, the intensity of stimulation and the area of the cortex being stimulated. Thus, transcranial magnetic stimulation preferentially activates different structures than transcranial electrical stimulation. In addition, the response to magnetic stimulation depends on the direction of the induced current in the brain, the waveform of the stimulating current, and the shape of the coil. Stimulation of the lower limb area of motor cortex recruits different elements than stimulation of the upper limb area. These differences occur because different structures in the motor cortex have a differential threshold to the different techniques of stimulation. We have had the opportunity to perform a series of direct recordings of the corticospinal volley evoked by the different techniques of transcranial stimulation from the epidural space of conscious patients with chronically implanted spinal electrodes. These recordings provide insights about the physiological basis of the excitatory and inhibitory phenomena produced by transcranial stimulation. PMID- 14744566 TI - Stochastic resonance and sensory information processing: a tutorial and review of application. AB - OBJECTIVE: To review the stochastic resonance phenomena observed in sensory systems and to describe how a random process ('noise') added to a subthreshold stimulus can enhance sensory information processing and perception. RESULTS: Nonlinear systems need a threshold, subthreshold information bearing stimulus and 'noise' for stochastic resonance phenomena to occur. These three ingredients are ubiquitous in nature and man-made systems, which accounts for the observation of stochastic resonance in fields and conditions ranging from physics and engineering to biology and medicine. The stochastic resonance paradigm is compatible with single-neuron models or synaptic and channels properties and applies to neuronal assemblies activated by sensory inputs and perceptual processes as well. Here we review a few of the landmark experiments (including psychophysics, electrophysiology, fMRI, human vision, hearing and tactile functions, animal behavior, single/multiunit activity recordings). Models and experiments show a peculiar consistency with known neuronal and brain physiology. A number of naturally occurring 'noise' sources in the brain (e.g. synaptic transmission, channel gating, ion concentrations, membrane conductance) possibly accounting for stochastic resonance phenomena are also reviewed. Evidence is given suggesting a possible role of stochastic resonance in brain function, including detection of weak signals, synchronization and coherence among neuronal assemblies, phase resetting, 'carrier' signals, animal avoidance and feeding behaviors. CONCLUSIONS: Stochastic resonance is a ubiquitous and conspicuous phenomenon compatible with neural models and theories of brain function. The available evidence suggests cautious interpretation, but justifies research and should encourage neuroscientists and clinical neurophysiologists to explore stochastic resonance in biology and medical science. PMID- 14744567 TI - Neonatal electrocortical brain activity and cerebral tissue oxygenation during non-acidotic, normocarbic and normotensive graded hypoxemia. AB - OBJECTIVE: Neonates are commonly exposed to isolated hypoxemic episodes. In order to identify the risk of this, we correlated cerebral oxygen delivery and electrocortical brain activity during isolated graded and repetitive hypoxemia in 1-week-old piglets. METHODS: Six halothane-anesthetized piglets were subjected to two episodes of graded hypoxemia of 45 min duration. The fractional concentration of inspired oxygen (FiO(2)) was stepwise decreased at 15 min intervals from 0.21 to 0.15, 0.10 and 0.05. A second identical hypoxemic event was induced after 1 h of normoxemia (FiO(2) 0.21). Mean arterial pressure (MAP) and pH were maintained at baseline values during the whole experiment. We measured near infrared spectroscopy parameters (cerebral oxidized cytochrome aa(3) (Cytaa3), total hemoglobin (tHb: oxy- +deoxyhemoglobin)) corresponding to cerebral blood volume (CBV), carotid blood flow (Q(car)), intra-arterial oxygen saturation (SaO(2)), and mean maximal EEG amplitude and relative spectral power. RESULTS: Delta (delta) power increased significantly and the EEG amplitude dropped below 10 and 5 microV at the end of the first and the second hypoxemic period (PaO(2) 2.68+/ 1.08 (P<0.05) and 2.87+/-0.58 kPa, respectively). Both EEG variables normalized during recovery (FiO(2) 0.21). Q(car), CBV and Cytaa3 were not changed. CONCLUSION: Acute isolated hypoxemia has to be sustained to induce neuronal hypofunction in normotensive animals. Hypoxic hypoxemia led to acute changes in neuronal activity, whereas cellular oxygenation remained unaffected. PMID- 14744568 TI - Impaired movement-related potentials in acute frontal traumatic brain injury. AB - OBJECTIVE: Focal brain lesions due to traumatic brain injury (TBI) do not only lead to functional deficits in the lesion area, but also disturb the structurally intact neuronal network connected to the lesion site. Therefore we hypothesized dysfunctions of the cortical motor network after frontal TBI. The movement related potential (MRP) is an EEG component related to voluntary movement consisting of the Bereitschaftspotential (BP), the negative slope (NS), and the motor potential (MP). The aim of our study was to demonstrate alterations in the movement related cortical network in the acute stage after TBI by comparing our patients' MRPs to those of a healthy control group. METHODS: EEGs of 22 patients with magnetic resonance imaging defined contusions of the prefrontal cortex were recorded within 8 weeks after TBI. We further recruited a healthy control group. The paradigm consisted of self-paced abductions of the right index finger. RESULTS: Compared to healthy controls, the BP in the patient group was significantly reduced and its onset delayed. Moreover, an enhanced contribution of the postrolandic hemisphere ipsilateral to the movement and a reduced contribution of the left frontal cortex, ipsilateral to the lesion in the majority of the patients, were observed during motor execution (MP). CONCLUSIONS: Anatomical connections between the prefrontal cortex and the supplementary motor area (SMA) are known to exist. We suggest that prefrontal lesions lead to reduced neuronal input into the SMA. This deficit in the preparatory motor network may cause the reduced BPs in our patients. Moreover, an increased need for attentional resources might explain the enhanced motor potentials during movement execution. In conclusion, we demonstrated altered MRPs in the acute stage after frontal TBI, which are a consequence of disturbed neuronal networks involved in the preparation and execution of voluntary movements. PMID- 14744569 TI - Individual analysis of EEG frequency and band power in mild Alzheimer's disease. AB - OBJECTIVE: This EEG study investigates the role of the cholinergic system, cortico-cortical connections, and sub-cortical white matter on the relationship between individual EEG frequencies and their relative power bands. METHODS: EEGs were recorded at rest in 30 normal elderly subjects (Nold), 60 mild Alzheimer disease (AD) and 20 vascular dementia (VaD) patients, comparable for Mini Mental State Evaluation scores (MMSE 17-24). Individual EEG frequencies were indexed by the theta/alpha transition frequency (TF) and by the individual alpha frequency (IAF) with power peak in the extended alpha range (5-14 Hz). Relative power was separately computed for delta, theta, alpha1, alpha2, and alpha3 bands, on the basis of the TF and IAF. RESULTS: Using normal subjects as a reference, VaD patients showed 'slowing' of alpha frequency (TF-IAF) and lower alpha2 power; Mild AD patients showed lower alpha2 and alpha3 power; delta power was higher in both AD and VaD patients; Theta power was higher only in VaD patients. CONCLUSIONS: Individual analysis of the alpha frequency and power can discriminate mild AD from VaD and normal elderly subjects. SIGNIFICANCE: This analysis may probe pathophysiological mechanisms causing AD and VaD. PMID- 14744570 TI - Magnetoencephalographic evidence of abnormal auditory processing in amyotrophic lateral sclerosis with bulbar signs. AB - OBJECTIVE: Amyotrophic lateral sclerosis (ALS) is characterized by progressive motor neuron damage that gives rise to muscle denervation. Besides motor neuron damage, conscious auditory processing appears to be impaired in ALS, whereas it has remained ambiguous whether preceding automatic auditory processing is abnormal in ALS and specifically in ALS with bulbar signs. METHODS: Auditory evoked fields (AEFs) to monaurally presented frequent and infrequent tones with stimulus intervals of 500 and 2500 ms were recorded with magnetoencephalography (MEG) from 10 ALS patients having bulbar signs and from 10 age-matched healthy subjects. RESULTS: The amplitudes of the P50m and N100m responses, which index automatic auditory processing underlying stimulus detection, were significantly increased and P50m latency was shortened in ALS patients. MMNm, which reflects memory-based auditory comparison process, was increased in amplitude in the patient group, whereas the MMNm latency was similar in both groups. AEF latency and amplitude values failed to correlate with the severity of ALS as measured by ALS Functional Rating Scale (ALSFRS). CONCLUSIONS: The present results suggest that auditory processing underlying stimulus detection, and subsequent memory based comparison processes are abnormal in ALS patients with severe bulbar signs. This might be due to cortical overactivity of excitatory neurotransmitter glutamate observed in ALS. PMID- 14744571 TI - Creutzfeldt-Jakob disease and non-convulsive status epilepticus: a clinical and electroencephalographic follow-up study. AB - OBJECTIVE: To describe the clinical and electroencephalographic findings from a confused elderly woman with Creutzfeldt-Jakob disease (CJD) that initially were compatible with the diagnosis of non-convulsive status epilepticus (NCSE). METHODS AND RESULTS: A 75-year-old right-handed woman was admitted to our hospital because of confusion and alteration of mental status. The two first electroencephalograms (EEGs) showed continuous diffuse spikes, rhythmic sharp waves and sharp-and-slow wave complexes which were completely abolished after the administration of 10 mg of intravenous diazepam. Over the following days, the clinical state of the patient was unmodified despite aggressive antiepileptic therapy. A third EEG revealed pseudo-periodic negative or positive-negative slow waves localised in the right frontal region. Subsequently, two consecutive EEGs showed continuous periodic generalised bi-triphasic complexes at a rate of 1 Hz, compatible with the diagnosis of CJD. Finally, the patient died, and postmortem examination was diagnostic of the sporadic form of CJD. CONCLUSIONS: Clinical and electroencephalographic features in the early stages of CJD may resemble NCSE. The administration of intravenous benzodiazepines and its clinical and electroencephalographic correlation, response to the antiepileptic therapy, and monitoring with serial EEG recordings may be helpful considerations in the differential diagnosis. PMID- 14744572 TI - Blink reflex abnormalities in Tourette syndrome. AB - OBJECTIVE: Tourette syndrome (TS) is a not uncommon disorder which represents the most complex manifestation of the spectrum of tic disorders, with onset during childhood or early adolescence. There are no definitive tests for diagnosis of TS. The objective of this study has been to demonstrate whether neurophysiological abnormalities of the blink reflex can be observed in patients affected with TS and correlate with the severity of TS. METHODS: We enrolled 17 patients with Tourette syndrome, diagnosed according to DSM IV Diagnostic Criteria, and 10 healthy volunteers. Tic severity was assessed using a self rating scale (Tourette Syndrome Symptom List, TSSL) and examiner ratings (Yale Global Tic Severity Scale (YGTSS), and Tourette-Syndrome Global Scale (TSGS)). The blink reflex was elicited by stimulating the supraorbital nerve in order to measure the early response (R1), homolateral and contralateral R2 (late) responses, amplitude of R1 and duration of R2. RESULTS: We observed a mean duration of R2 significantly longer in the patient group than in the control group (P<0.01, Student t test), without any statistically significant differences of R1 and R2 latencies and of R1 amplitude between the patient group and the control group. Correlations between changes in clinical rating scores and R2 duration were tested by simple linear regression analysis, which has not demonstrated a significant correlation between TSSL scores, clinical rating scores (measured by TSGS and YGTSS) and duration of R2. CONCLUSIONS: A pattern as to excitability of the blink reflex can be a frequent abnormality in TS patients, not correlated with its severity. PMID- 14744573 TI - Flumazenil does not affect intracortical motor excitability in humans: a transcranial magnetic stimulation study. AB - OBJECTIVE: The motor cortex may be subject to tonic inhibitory drive. One inhibitory mechanism is supported by activity at benzodiazepine (BZP) receptors. In this study we investigate whether or not the BZP antagonist, flumazenil, increases cortical motor excitability in humans. METHODS: Eight healthy subjects received a 1 mg intravenous (i.v.) loading dose of flumazenil followed by a 0.5 mg i.v. infusion over the next 30 min. Before, during, and 1 h after flumazenil infusion, we measured cortical motor excitability using transcranial magnetic stimulation (TMS). This included resting motor threshold (rMT), paired-pulse measurements of intracortical inhibition and facilitation (ICI and ICF), recruitment curve (RC), and silent period (SP). We also measured F response and compound muscle action potential (CMAP) with peripheral nerve stimulation. The study was carried out using a randomized, double-blind crossover design controlled with a saline infusion. RESULTS: None of the measures of cortical or peripheral excitability were significantly affected by drug administration. CONCLUSIONS: Our findings suggest that flumazenil has no effect on cortical motor excitability in normal humans. SIGNIFICANCE: There does not appear to be any tonic activity at benzodiazepine receptors in the normal resting human motor cortex. PMID- 14744574 TI - Subtle hemispheric asymmetry of motor cortical inhibitory tone. AB - OBJECTIVE: To test whether a novel paired transcranial magnetic stimulation (TMS) protocol (J Physiol 545.1 (2002) 153) detects hemispheric differences in motor cortical inhibition. METHODS: Nine right-handers and 8 left-handers participated. Focal paired TMS was applied to the hand area of the dominant (M1-D) or non dominant motor cortex (M1-ND). Motor evoked potentials (MEP) were recorded from the relaxed contralateral abductor digiti minimi. The first (S1) and second pulse (S2) were separated by 1.5 or 2.1 ms. Nine stimulus intensities of S1 and S2 (i.e. 9x9 intensity conditions) ranging from 60 to 140% of resting motor threshold (RMT) were tested. The interaction between S1 and S2 was expressed by MEP(S1+S2)/(MEP(S1)+MEP(S2))*100%. Values below and above 100% indicate short interval intracortical inhibition (SICI) and facilitation (SICF), respectively. RESULTS: In right-handers, RMT was lower, SICI was present with fewer intensity conditions and the magnitude of SICI was less in M1-D than M1-ND. No hemispheric asymmetry was found for SICF. Left-handers showed no hemispheric difference for any of these measures. CONCLUSIONS: Findings suggest that, in right-handers, M1-D is controlled by less inhibitory tone than M1-ND. This may put the M1-D to an advantage for processes that are associated with a reduction of SICI, such as voluntary activation and use-dependent plasticity. PMID- 14744575 TI - Cortical control of voluntary blinking: a transcranial magnetic stimulation study. AB - OBJECTIVE: To investigate cortical regions related to voluntary blinking. METHODS: Transcranial magnetic stimulation (TMS) was applied to the facial motor cortex (M1) and the midline frontal region (Fz) in 10 healthy subjects with eyes opened and closed. Motor-evoked potentials were recorded from the orbicularis oculi (OOC), orbicularis oris (OOR), abductor digiti minimi and tibialis anterior using surface and needle electromyography electrodes. Facial M waves and blink reflex were measured using supramaximal electrical stimulation of the facial and supraorbital nerves. RESULTS: TMS at Fz elicited 3 waves in OOC with no response in other tested muscles except for the early wave in OOR. Facial M1 stimulation produced only early and late waves. Because of their latencies, shapes, and relationship to coil position and stimulation intensity, early and late waves appeared to be analogous to the facial M wave and R1 component of the blink reflex. The intermediate wave at 6-8 ms latency was elicited in OOC by Fz stimulation with eyes closed. CONCLUSIONS: Since its latency matches the central conduction time of other cranial muscles and it has characteristic of muscle activation-related facilitation, the intermediate wave is presumably related to cortical stimulation. This result provides evidence that the cortical center for the upper facial movements, including blinking, is not principally located in the facial M1, but rather in the mesial frontal region. PMID- 14744576 TI - Stimulus-response properties of motor system in patients with cerebellar ataxia. AB - OBJECTIVE: The aim of the study was to examine the stimulus-response properties of the excitatory and inhibitory components of corticospinal projections at rest and during voluntary contraction in cerebellar patients. METHODS: We investigated motor evoked potential (MEP) and cortical silent period recruitment curves in response to increasing intensities of transcranial magnetic stimulation in 8 patients with 'pure' cerebellar syndromes and in 14 age-matched controls. The transcranial magnetic stimulation intensity was increased from 90 to 180% of the resting motor threshold. MEP recruitment curves were recorded at rest and during voluntary contraction in the right abductor pollicis brevis muscle. RESULTS: No statistical differences were found between patients and controls in MEP recruitment curves in either the resting or active condition. A significant difference was found between patients and controls in the cortical silent period threshold (patients: 33.2+/-3.4% of maximal stimulator output; controls 39.4+/ 3.2%; P=0.01) and recruitment curve, the duration of the cortical silent period being longer in patients at transcranial magnetic stimulation intensities ranging from 90 to 130% of the resting motor threshold (patients: 135-191 ms; controls: 53-158 ms). No changes were found in the silent period evoked by peripheral nerve stimulation. CONCLUSIONS: Inhibitory components of corticospinal projections were recruited with a lower threshold in patients. No abnormalities were found in the recruitment of the excitatory networks. Our data show a prevalence of inhibitory phenomena in the motor cortex of cerebellar patients. These findings would appear to be specific to cerebellar diseases and are the opposite of those previously documented in movement disorders such as dystonia and Parkinson's disease. Our results suggest that the cerebellum and the basal ganglia may counteract each other in modulating the level of motor system excitability. PMID- 14744577 TI - Triple stimulation technique (TST) in amyotrophic lateral sclerosis. AB - OBJECTIVE: The authors studied amyotrophic lateral sclerosis (ALS) patients using triple stimulation technique (TST) to detect upper motor neuron (UMN) involvement. METHODS: Nineteen ALS patients (aged 45-72 years) were enrolled in the study. According to the El Escorial criteria, 6 diagnoses were suspected or possible, 6 probable, and 7 definite. Patients were examined clinically, with conventional (single-pulse) transcranial magnetic stimulation (TMS), and with TST (on one side only). RESULTS: Among the whole group of patients, TST appeared to be more sensitive than conventional TMS techniques. In particular among suspected/possible ALS patients, TST area ratio was pathologic in 100%, while single-pulse TMS was abnormal only in 50% of cases. Overall, the use of TST area ratio was more sensitive than the analysis of TST amplitude ratio. CONCLUSIONS: The results suggest that TST might be more sensitive and useful in the diagnosis of subclinical UMN involvement than conventional TMS techniques, even if TST is performed on one side only. PMID- 14744578 TI - Effects of a go/nogo task on event-related potentials following somatosensory stimulation. AB - OBJECTIVE: We investigated the effects of a go/nogo task on event-related potentials (ERPs) evoked by somatosensory stimuli. METHODS: ERPs following electrical stimulation of the second (go stimulus) or fifth (nogo stimulus) left handed digit were recorded from 9 subjects. The recordings were conducted in 3 conditions: Control, Count and Movement. The subjects were instructed to count the go stimuli silently in Count, and respond to the go stimuli by grasping right hands in Movement. Go and nogo stimuli were presented at an even probability. RESULTS: N140 was recorded in all conditions and P300 in Count and Movement. The mean amplitudes of the nogo stimuli in the interval 140-200 msec and nogo-N140 amplitude were significantly more negative than those of the go stimuli in Count or Movement. Nogo-P300 was larger in amplitude than go-P300 in Movement but not Count. The effect of P300 was applied to Fz and Cz, but not at Pz. CONCLUSIONS: In the present study, effects of a somatosensory go/nogo task on ERPs were investigated, and our findings were very similar to those of previous studies using visual and auditory go/nogo tasks. Therefore, we suggest that cortical activities relating to go/nogo tasks are not dependent on sensory modalities. SIGNIFICANCE: The present study showed for the first time the go/nogo effects on somatosensory-evoked ERPs. These effects were similar to those in visual and auditory ERP studies. PMID- 14744579 TI - Predictive validity of event-related potentials (ERPs) in relation to the directed forgetting effects. AB - OBJECTIVE: A directed forgetting paradigm (word method) was used to assess the relationships between the event-related potentials (ERPs) recorded during the study phase and the subsequent forgetting effects. METHODS: In the study phase 100 words were presented each followed by either an instruction to remember (R) or to forget (F). Then these 100 words, together with another 100 new words, were presented and subjects had to perform an old/new decision task. The forgetting effect was considered as the difference between the rate of R- and F-words correctly recognised. Based on this difference, the whole sample was divided into two 10-subject groups with high and low forgetting effect. RESULTS: From 300 to 600 msec post-instruction differences in ERPs between R and F instructions were similar in both groups. In contrast, in the 100-300 msec epoch, ERP differences only appeared in the high-effect group and consisted of an enhanced positive activity elicited by F instruction at frontal and prefrontal areas and a larger positivity associated to R instruction at parietal area. CONCLUSIONS: The findings suggest that the processing of the word is kept on stand-by until the instruction is provided. Instruction to remember would reactivate this processing, which would mainly involve the parietal area, whereas instruction to forget appears to trigger frontal and prefrontal mechanisms trying to inhibit such processing. PMID- 14744580 TI - Habituation in a simple repetitive motor task: a study with movement-related cortical potentials. AB - OBJECTIVE: The movement-related cortical potential (MRCP) attenuates over the right hemisphere during the performance of a complex motor sequence. Because subjects have to learn complex sequences, earlier research has attributed this decrease of negativity to motor learning. The aims of this study were (1) to examine whether such an attenuation also occurs in the course of a repetitive motor task so simple that learning is not required, and (2) whether there are any transfer effects when subjects change from one to the other hand. METHODS: MRCPs were recorded in 33 right-handed healthy subjects. In a counterbalanced crossover design, subjects made 100 self-paced unilateral button presses with their left or right index finger, and then continued on the other side of the body for another 100 movements. Tasks were performed with the same speed during the entire experiment. RESULTS: (1) The amplitude of the MRCP attenuated over the right hemisphere in the course of the tasks irrespective of the side of movement. (2) The amplitude of the MRCP decreased across the entire scalp until a minimum level was reached at the end of the earlier sequence, and then increased slightly from this low level during the following sequence made after a switch of hand. CONCLUSIONS: The right hemisphere may predominate motor activation or attentional demands directed towards movement execution or somatosensory inputs even in very simple motor tasks. The decrease and subsequent increase of negativity during the first and second sequence, respectively, may reflect the subjects' changing arousal. PMID- 14744581 TI - Language-related brain activity revealed by independent component analysis. AB - OBJECTIVE: When an individual engages in a cognitive task, a multitude of diverse processes are activated in his/her brain and it is reasonable to assume that multiple brain sources are simultaneously active at any one time. Magnetoencephalographic (MEG) data recorded in such circumstances provide a picture of spatial distribution and time course of the sum of the magnetic fields generated by all these sources. Thus, the experimenter faces the challenge of separating the multiple contributions to the total recorded signal before attempting a localization of their sources and studying their functional roles. METHODS: We describe in this paper how independent component analysis of MEG data collected in a word/pseudo-word reading experiment elegantly solves this problem. RESULTS: Using a few statistical assumptions, independent component analysis resolved simultaneously active brain sources in the right-frontal, left-parietal and left-frontal areas, all showing well defined dipolar field distributions. DISCUSSION: We describe the characteristics of these contributions and discuss the language-related functional roles that appear to be associated to some of the independent sources. We report in particular on one source, localized near Broca's area, which showed to be affected by reading words but not pseudo-words. PMID- 14744582 TI - Clozapine, but not typical antipsychotics, correct P50 suppression deficit in patients with schizophrenia. AB - OBJECTIVE: To find out if there is a difference in P50 suppression between patients using typical antipsychotic drugs and those using clozapine, as well as to confirm the findings of abnormal P50 suppression in patients with schizophrenia, when compared to healthy volunteers. METHODS: Fifty patients with schizophrenia and 25 healthy volunteers were divided into 3 groups: group 1 - patients using typical antipsychotics; group 2 - patients using clozapine; group 3 - controls. Before the examination, all patients were interviewed by a psychiatrist using the Brief Psychiatry Rating Scale (BPRS). RESULTS: The average S2/S1 ratio was 0.82+/-0.45 in group 1, 0.57+/-0.41 in group 2, and 0.44+/-0.27 in group 3 (P=0.003). Statistical analysis showed a significant difference when the results of group 1 were compared to those of groups 2 (P=0.045) and 3 (P=0.001). There was no significant difference between groups 2 and 3 (P=0.182). There was a significant difference in the S1-S2 difference only between groups 1 and 3 (P=0.007), but a non-significant trend towards a similar difference was found between groups 1 and 2 (P=0.067). There was no correlation between the BPRS values and any P50 parameter. CONCLUSIONS: The suppression of P50 among patients using clozapine was significantly greater than that obtained in patients using typical antipsychotics. SIGNIFICANCE: This study confirms, in a more evident way, the improvement of the suppression of P50 potential in schizophrenics using clozapine. Additionally, it discusses the physiopathological mechanism involved. PMID- 14744583 TI - EEG differences in children as a function of resting-state arousal level. AB - OBJECTIVE: This study aimed to explore the basis of a theoretical position which has major impact in the current literature on attention-deficit/hyperactivity disorder (AD/HD) - that the ratio of theta to beta power in the EEG (the theta/beta ratio) represents central nervous system (CNS) arousal. METHODS: Resting state topographic EEG activity was investigated as a function of CNS arousal in normal right-handed boys. Arousal was defined in terms of electrodermal activity, which has a long history in Psychology as a measure of CNS arousal. RESULTS: Relative delta, theta, and beta power, and the theta/beta ratio, failed to differ significantly between age-matched groups which differed markedly in skin conductance level and non-specific electrodermal fluctuations. The high-arousal group showed significantly lower levels of relative alpha power, particularly in posterior and hemispheric regions, with higher alpha frequencies in these areas. CONCLUSIONS: These data fail to support the theoretical linkage between the theta/beta ratio and CNS arousal. Further work is needed to disentangle the different correlates of arousal and task-related activation, particularly in cognitive and attentional-processing terms. SIGNIFICANCE: The outcomes of this study, in addition to clarifying the nature of EEG markers of CNS arousal, have important implications for our understanding of AD/HD, as they require re-evaluation of current models of the disorder. PMID- 14744584 TI - Developmental changes in ERP correlates of spoken word recognition during early school years: a phonological priming study. AB - OBJECTIVE: We investigated event-related potential (ERP) correlates of developmental changes in spoken word recognition during early school years. We focused on implicit processing of word onsets as this may change considerably due to vocabulary growth and reading acquisition. METHODS: Subjects were pre schoolers (5-6 years), beginning readers (7-8 years) and adults. Two experiments examined phonological onset priming effects on ERP measures in an auditory lexical decision task. Primes were words (Expt. 1) or non-words (Expt. 2). RESULTS: ERPs elicited by words showed clear developmental changes in ERP latency and morphology, especially with regard to early negativities like the N1. Prominent priming effects were an enhanced N400 amplitude due to alliterating word primes in beginning readers, and typical phonological N400 reductions due to alliterating non-word primes in all groups. Priming further led to opposite early effects with word primes (N1/N(200) reduction) vs. non-word primes (P1/N1 enhancement) in all groups. CONCLUSIONS: These changes in ERP morphology and priming effects suggest that the lexical system undergoes substantial restructuring at the level of phonological processing and representation. In particular, our results indicate distinct processing of word onsets in beginning readers, confirming that vocabulary growth and the acquisition of reading may critically contribute to the formation of a fully segmental lexical system. PMID- 14744585 TI - Somatosensory evoked potentials (SEPs) recorded from deep brain stimulation (DBS) electrodes in the thalamus and subthalamic nucleus (STN). AB - OBJECTIVE: To examine the location of deep brain stimulation (DBS) electrode somatosensory evoked potentials (SEPs) and determine the generators of the median nerve SEPs recorded in thalamus and subthalamic nucleus (STN). METHODS: SEPs were recorded from contacts of DBS electrodes and microelectrodes in thalamus and STN to establish the latencies of N13, N18 and N20 in 24 patients (8 tremor, 4 chronic pain, 12 Parkinson disease) undergoing chronic DBS. RESULTS: A large SEP with a mean latency of 17.9+/-1.7 ms was recorded from thalamic contacts. Phase reversal occurred at the horizontal level of the anterior commissure-posterior commissure line. Smaller potentials with similar latency but no reversal could be recorded from STN electrodes. CONCLUSIONS: We propose that the thalamic SEP is generated by excitatory post-synaptic potentials in sensory relay neurons in nucleus ventrocaudalis. A small potential in STN at a similar latency, may be due to volume conduction from thalamus. Intraoperative and postoperative SEP recordings from DBS electrodes could be used to determine the optimal position of the contacts relative to the sensory pathways and the choice of contacts for chronic stimulation. PMID- 14744586 TI - Is there training-dependent reorganization of digit representations in area 3b of string players? AB - OBJECTIVE: The digit representations in area 3b were studied to examine whether there is training-dependent reorganization in string players. METHODS: Somatosensory evoked magnetic fields were recorded following electrical stimulation of digits 1 (D1), 2 (D2) and 5 (D5) of both hands in 8 string players and of the left hand in 12 control subjects. The N20m and P30m responses, and high-frequency oscillations (HFOs) were separated by 3-300 Hz and 300-900 Hz bandpass filtering. RESULTS: The dipole locations on the coronal plane and strengths of D1, D2 and D5, and D1-D5 cortical distance estimated at the peak of N20m or P30m did not differ between left and right hand in string players or between left hand in string players and controls. On the other hand, the dipole locations of D2 estimated from N20m and P30m and of D1 from N20m were significantly anterior, the D2-D5 distance from P30m longer, and the number of HFO peaks larger for D5 in string players than controls. CONCLUSIONS/SIGNIFICANCE: With strong mutual competition among the fingering digits, the scale of reorganization should be much smaller as compared with the competition-free denervation-induced reorganizations. Taken together, the training-dependent reorganization of somatosensory cortex in string players is manifest not only in the enlarged cortical representation but also in the enhanced HFOs presumably representing activity of the fast-spiking interneurons. PMID- 14744587 TI - Somatosensory evoked potentials and magnetic fields elicited by tactile stimulation of the hand during active and quiet sleep in newborns. AB - OBJECTIVE: Our objective was to characterize the effects of sleep stages on tactile somatosensory evoked responses in full-term newborns. METHODS: Somatosensory evoked potentials (SEPs) and magnetic fields (SEFs) to tactile stimulation of the tip of the index finger and/or thenar eminence were measured from 14 healthy newborns. The stimulus was a gentle tap produced by a moving membrane driven by an air-pressure pulse. RESULTS: SEPs and SEFs to tactile stimulation of the skin were similar in waveform and latency to SEPs known to be produced by electrical stimulation of the fingertip of neonates. The two most distinguishable positive deflections of SEPs, P1 and P2, within 300 ms of the stimulation, and their magnetic counterparts were clearly smaller in active compared to quiet sleep. CONCLUSIONS: Our study demonstrates for the first time that it is possible to record SEFs in neonates, and that clear late cortical somatosensory responses are produced by tactile stimulation. In addition, the effect of sleep stage on these responses indicates differences in the processing of the incoming information, at least in the somatosensory modality, in active and quiet sleep. SIGNIFICANCE: Tactile stimulation may be useful as a completely non-invasive technique for studying the physiology of the somatosensory system in neonates. Methodologically, since the effect of sleep stage is profound, one must carefully monitor the sleep stages in studies of event-related responses in newborns, or else this effect may confound the phenomena being studied. PMID- 14744588 TI - Effect of transcranial DC sensorimotor cortex stimulation on somatosensory evoked potentials in humans. AB - OBJECTIVE: To study the after-effect of transcranial direct current stimulation (tDCS) over the sensorimotor cortex on the size of somatosensory evoked potentials (SEPs) in humans. METHODS: SEPs were elicited by electrical stimulation of right or left median nerve at the wrist before and after anodal or cathodal tDCS in 8 healthy subjects. tDCS was applied for 10 min to the left motor cortex at a current strength of 1 mA. RESULTS: Amplitudes of P25/N33, N33/P40 (parietal components) and P22/N30 (frontal component) following right median nerve stimulation were significantly increased for at least 60 min after the end of anodal tDCS, whereas P14/N20, N20/P25 (parietal components) and N18/P22 (frontal component) were unaffected. There was no effect on SEPs evoked by left median nerve stimulation. Cathodal tDCS had no effect on SEPs evoked from stimulation of either arm. CONCLUSIONS: Anodal tDCS over the sensorimotor cortex can induce a long-lasting increase in the size of ipsilateral cortical components of SEPs. SIGNIFICANCE: tDCS can modulate cortical somatosensory processing in humans and might be a useful tool to induce plasticity in cortical sensory processing. PMID- 14744589 TI - Visual electrophysiological screening in diagnosing infants with congenital nystagmus. AB - OBJECTIVE: To determine the value of visual electrophysiological screening in evaluating retinal and postretinal visual pathway function in infants with congenital nystagmus. METHODS: In 28 infants with congenital nystagmus electroretinogram (ERG) was recorded with skin electrodes and, since the infants were alert, visual evoked potentials (VEP) were simultaneously recorded. The first recording was performed before the age of 1 year (age 2-11 months, mean age 6.8 months). Follow-up was performed between the ages of 7 months and 6 years (mean age 2.7 years). ERG was detected to flash stimulus and VEP to flash and/or pattern-reversal stimulus. RESULTS: In infants less than 1 year of age, retinal dysfunction was recognised in 36% of infants who had been diagnosed as Leber's congenital amaurosis, achromatopsia and retinal dystrophy; postretinal dysfunction was recognised in 32% of infants who had been diagnosed with ocular albinism, optic nerve hypoplasia and neurological nystagmus; retinal or postretinal function was normal in 7% of infants that had been diagnosed as congenital idiopathic nystagmus, while retinal function was normal and postretinal function was undefined in 25% of infants that had been diagnosed as optic nerve hypoplasia, neurological nystagmus and congenital idiopathic nystagmus. At follow-up the findings were: retinal dysfunction in 36%, postretinal dysfunction in 50% and normal retinal and postretinal function in 14% of children. All those children with normal retinal and postretinal function had been diagnosed as congenital idiopathic nystagmus. CONCLUSIONS: Visual electrophysiological screening of infants with congenital nystagmus can establish or exclude retinal and postretinal pathway dysfunction. Therefore simultaneous skin ERG and VEP which is a non-invasive approach for infants enables an objective means for identifying the basis of congenital nystagmus, thereby assisting in its classification. PMID- 14744590 TI - Evoked K-complex generation: the impact of sleep spindles and age. AB - OBJECTIVE: It has been proposed that spindles and spontaneous K-complexes reflect two sides of a coin, with the spindle reflecting an inhibitory microstate and the K-complex reflecting an excitatory or aroused microstate [Physiol Behav 1993;54(4):795]. This hypothesis predicts that the presence of a sleep spindle at the time of stimulus presentation would decrease the likelihood of a K-complex being elicited by that stimulus. The present study sought to test this hypothesis in young and elderly subjects. METHODS: Ten young and 7 elderly adults who were neurologically healthy and free from medications spent one night in the sleep laboratory. EEG was recorded from 6 gold plate electrodes (Fz, FCz, Cz, CPz, Pz and O2) referenced to A1+A2. Tone clicks (1000 Hz) at 80 dB above measured awake detection thresholds were presented binaurally either during a spindle (SP+) or in the absence of a spindle (SP-). This was achieved by viewing a central EEG channel filtered to pass only the frequencies between 12 and 14 Hz. Trials were further classified based on whether (KC+) or not (KC-) they produced a K-complex. K-Complex probability and the amplitude and latency of the N550 component of the averaged evoked potential for KC+ trials were assessed using a two-way analysis of variance with main effects of age and spindle presence/absence. RESULTS: There were significant reductions in K-complex probability and N550 amplitude and a significant increase in N550 latency, as a function of age. However, no variable displayed a significant effect of spindle presence/absence, or an age x spindle interaction effect. CONCLUSIONS: The data failed to support the hypothesis that sleep spindles are antagonistic to the production of K-complexes, both in terms of the likelihood of K-complexes being elicited or in their amplitude when elicited (N550). The absence of spindle effects on K-complex generation argues against them being two sides of a coin and supports the notion of K-complexes having an extra-thalamic or non-specific generation mechanism. PMID- 14744592 TI - Assessing somatosensory evoked potential (SEP) generators by human intracranial recordings. PMID- 14744591 TI - Canine electroencephalographic recording technique: findings in normal and epileptic dogs. AB - OBJECTIVE: The absence of a standardized recording technique and of normal reference patterns may have hindered the use of experimental electroencephalography (EEG) in dogs and in veterinary medicine as well. Therefore, the aim of the present work was to develop a recording technique for brachycephalic, mesocephalic and dolichocephalic breeds of dogs that would standardize the recording parameters, including the electrical activity of the rhinencephalon and prorean gyrus, and to prove its feasibility for detecting abnormal activity. METHODS: We accomplished this task by employing 12 recording electrodes. Three different doses of xylazine (0.5, 1 and 1.5 mg/kg) were used to determine which gave enough sedation to obtain a recording free of artifact. Fifty normal dogs were studied; for comparison 9 epileptic dogs were also explored. RESULTS: We found specific anatomical skull sites where the EEG recording electrodes should be attached; the location of these sites is, in practical terms, the same for every dog, regardless of their skull type. A dose of 1 mg/kg xylazine was chosen for restraining. Normal dogs displayed predominant 6-12 Hz waves which could be recognized all over the cortex. Epileptic dogs were individualized because of the appearance of paroxysmal discharges, composed of spikes, sharp and anomalous sinusoidal waves which usually started in one area, spreading to the whole cortex later on, in the clinical interictal periods. CONCLUSIONS: It was possible to obtain more extensive electrophysiological coverage of the canine cerebral cortex, including the rhinencephalon and prorean gyrus, than previously described in the literature. Intramuscular or subcutaneous xylazine, at a dose of 1 mg/kg, proved to produce the best restraint for recordings. The technique allowed to distinguish abnormal activity in epileptic dogs. PMID- 14744595 TI - Antiproliferative mechanisms of raxofelast (IRFI-016) in H2O2-stimulated rat aortic smooth muscle cells. AB - Reactive oxygen species-mediated cellular injury is involved in the pathogenesis of many diseases, including those affecting the cardiovascular system, such as myocardial ischemia-reperfusion injury, inflammation, and atheroscleosis. Raxofelast (IRFI-016; (+/-)-5-acetoxy-2, 3-dihydro-4, 6, 7-trimethyl-2-benzofuran acetic acid) was designed with the aim of maximizing the antioxidant potency of phenols chemically related to vitamin E. The antioxidant activity of raxofelast has been convincingly demonstrated in several in vitro studies and in various models of ischemia-reperfusion injury. In this study, the antiproliferative effects of raxofelast were investigated to determine whether transduction signals and protooncogenes are affected in H(2)O(2)-stimulated rat aortic smooth muscle cells. In a tetrazolium-based colorimetric assay, the proliferation of rat aortic smooth muscle cells was increased by 3-fold in 0.1% fetal bovine serum/Dulbecco's modified Eagle's medium (DMEM) containing 500 microM H(2)O(2), indicating that exogenous 500 microM H(2)O(2) was a growth stimulator of rat aortic smooth muscle cells. Exogenous H(2)O(2) significantly activated extracellular signal-regulated kinases (ERKs) activity within 30 min and raxofelast inhibited the ERKs activation dose dependently in 500 microM H(2)O(2)-stimulated rat aortic smooth muscle cells (IC(50): 200 microM). Raxofelast reduced the intracellular reactive oxygen species generated by exogenous H(2)O(2) in a dose-dependent manner. In 500 microM H(2)O(2)-stimulated rat aortic smooth muscle cells, raxofelast dramatically attenuated the activation of mitogen-activating protein kinase (MAPK)/ERK kinase 1, 2 (MEK1,2) and protein kinase C (PKC) without affecting Ras expression. Induction of c-myc mRNA was significantly reduced dose dependently up to 100 microM by raxofelast in concentrations. These data indicate that the antiproliferative effects of raxofelast in H(2)O(2)-stimulated rat aortic smooth muscle cells may involve the suppression of intracellular reactive oxygen species formation and the inhibition of ERKs by inactivation through PKC and MEK1,2 and down-regulation of c-myc expression, regardless of Ras activation. PMID- 14744596 TI - Molecular cloning and pharmacological characterization of the guinea pig 5-HT1E receptor. AB - The human 5-HT(1E) receptor gene was cloned more than a decade ago. Little is known about its function, and there have been no reports of its existence in the genome of small laboratory animals. In this study, attempts to clone the 5-HT(1E) gene from the rat and mouse were unsuccessful. In fact, a search of the mouse genome database revealed that the 5-HT(1E) receptor gene is missing from the mouse genome. However, the 5-HT(1E) gene was cloned from guinea pig genomic DNA and was characterized. The guinea pig 5-HT(1E) receptor gene encodes a protein of 365 amino acids. It shares 88% (nucleic acid) and 95% (amino acid) homology with the human receptor. The guinea pig 5-HT(1E) receptor showed similar pharmacology to the human 5-HT(1E) receptor in radioligand binding assays. Serotonin (5 hydroxytryptamine, 5-HT) dose-dependently stimulated [35S]GTPgammaS binding to the guinea pig 5-HT(1E) receptor with an EC(50) of 13.6+/-1.92 nM, similar to that of the human 5-HT(1E) receptor (13.7+/-1.78 nM). Activation of the guinea pig 5-HT(1E) receptor was also achieved by ergonovine, alpha-methyl-5-HT, 1 naphthylpiperazine, methysergide, tryptamine, and 1-(2,5-dimethoxy-4-iodophenyl) 2-aminopropane (DOI). Methiothepin exhibited antagonist activity. Quantitative real-time polymerase chain reaction (qRT-PCR) analysis showed that 5-HT(1E) mRNA was present in the guinea pig brain with the greatest abundance in the hippocampus, followed by the olfactory bulb. Lower levels were detected in the cortex, thalamus, pons, hypothalamus, midbrain, striatum, and cerebellum. Our current study marks the first identification of the 5-HT(1E) receptor gene in a commonly used laboratory animal species. This finding should allow the elucidation of the receptor's role(s) in the complex coordination of central serotonergic effects. PMID- 14744597 TI - Acute actions of 3 alpha-hydroxy-tibolone on factors influencing contraction in guinea-pig ventricular myocytes. AB - We investigated whether one of the main estrogenic metabolites of the postmenopausal agent, tibolone (Org OD14), exerts direct cardiac actions in a similar way to 17beta-estradiol. 3alpha-OH-tibolone (40 microM) decreased both cell shortening and Ca2+ transient amplitude of field-stimulated (1 Hz, 37 degrees C) guinea-pig ventricular myocytes. These effects were still observed in cells that had been incubated with the specific estrogen receptor antagonist, ICI 182,780 (C(32)H(47)F(5)O(3)S), suggesting an estrogen receptor-independent mechanism of action. In addition, 3alpha-OH-tibolone inhibited the L-type Ca2+ current and shortened action potential durations (APD). This mechanism may contribute to a potential cardiovascular action of tibolone. PMID- 14744598 TI - Suppressive effects of FR167653, an inhibitor of p38 mitogen-activated kinase, on calreticulin mRNA expression induced by endoplasmic reticulum stresses. AB - Several endoplasmic reticulum chaperones are simultaneously transactivated in response to various forms of endoplasmic reticulum stresses. Calreticulin is one such chaperone. We here show that the compound FR167653 [1-[7-(4-fluorophenyl) 1,2,3,4-tetrahydro-8-(4-pyridyl)pyrazolo[5,1-c][1,2,4]triazin-2-yl]-2 phenylethanedione sulfate monohydrate] suppresses the transactivation of calreticulin following endoplasmic reticulum stress. FR167653, like SB203580 [4 (4-fluorophenyl)-2-(4-methylsulfinylphenyl)-5-(4-pyridyl)-imidazole], has been reported to inhibit p38 mitogen-activated kinase (p38 MAPK). In this study, FR167653 concentration-dependently inhibited the up-regulation of the calreticulin mRNA level following an endoplasmic reticulum stress induced by thapsigargin in human embryonic kidney 293 (HEK293) cells and rat phechromocytoma PC12 cells. The compound concentration-dependently suppressed the transactivation of luciferase by thapsigargin in a reporter assay with a calreticulin promoter luciferase conjugated reporter vector. SB203580 also significantly suppressed the transactivation of calreticulin by thapsigargin. Therefore, FR167653 regulated the mRNA expression of calreticulin at the transcriptional level without affecting the stability of the mRNA, as well as via inhibition of p38 MAPK activated by thapsigargin. FR167653 also inhibited the transactivation of calreticulin stimulated by two other endoplasmic reticulum stress inducers, tunicamycin and A23187. Moreover, the inhibitory action of the compound on the transactivation was observed in other cell lines. The calreticulin promoter region includes three sequential cis-acting endoplasmic reticulum stress response elements (ERSEs). As each of these ERSEs was sequentially deleted, there was an increasing loss of the transactivation by thapsigargin or tunicamycin. FR167653 inhibited the transactivation in all the reporter plasmid constructs containing the calreticulin promoter region with an ERSE/ERSEs. In conclusion, FR167653 is the first compound shown to inhibit the transactivation of calreticulin following various endoplasmic reticulum stresses. The suppressive effects of the compound were considered to be due to an inhibition of the signaling leading to ERSEs activation in the calreticulin promoter region via an inhibition of p38 MAPK, which is activated by endoplasmic reticulum stresses. PMID- 14744599 TI - Analysis of the analgesic interactions between ketorolac and tramadol during arthritic nociception in rat. AB - The potential advantage of using combination therapy is that analgesia can be maximized while minimizing the incidence of adverse effects. In order to assess a possible synergistic antinociceptive interactions, the antinociceptive effects of ketorolac tromethamine, p.o., a nonsteroidal anti-inflammatory drug (NSAID), and tramadol hydrochloride, p.o., an atypical opioid analgesic, administered either separately or in combination, were determined using a rat model of arthritic pain. The data were interpreted using the surface of synergistic interaction (SSI) analysis and an isobolographic analysis to establish the nature of the interaction. 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. Female rats received orally ketorolac alone (0.18, 0.32, 0.56, 1.0, 1.78, 3.16, and 5.62 mg/kg), tramadol alone (3.16, 5.62, 10.0, 17.78, 31.62, 56.23, and 100.0 mg/kg), or 24 different combinations of ketorolac plus tramadol. Ten combinations exhibited various degrees of potentiation of antinociceptive effects (17.78 mg/kg tramadol with either 0.18, 0.32, or 0.56 mg/kg ketorolac; 10.0 mg/kg tramadol with either 0.18, 0.32, 0.56, or 1.8 mg/kg ketorolac; 5.6 mg/kg tramadol with either 0.32 or 0.56 mg/kg ketorolac; and 3.16 mg/kg tramadol with 0.32 mg/kg ketorolac), whereas the other 14 combinations showed additive antinociceptive effects. Three combinations of ketorolac+tramadol (1.0+17.78, 1.78+10, and 1.78+17.78, mg/kg respectively) produced the maximum antinociceptive effects, and two combinations (0.32+10.0 and 0.56+10.0 mg/kg, respectively) presented effects of high potentiation (P<0.001). This combination caused gastric injuries less severe than those observed with indomethacin. The synergistic antinociceptive effects of ketorolac and tramadol were important and suggest that combinations with these drugs may have clinical utility in pain therapy. PMID- 14744600 TI - Alleviation of ischemic neuronal damage by histamine H2 receptor stimulation in the rat striatum. AB - Transient ischemia was produced for 15 min by occlusion of the middle cerebral artery in halothane-anesthetized rats, and changes in the extracellular concentrations of neurotransmitter monoamines and amino acids were examined in the striatum. The occlusion produced marked increases in the extracellular concentrations of both dopamine and glutamate in the striatum in the saline injected control group, the peak values being 148 and 5.2 times those before ischemia, respectively. Preischemic administration of histamine (200 nmol, i.c.v.) suppressed the increase in dopamine and glutamate levels during ischemia, the peak values being 38% and 40% of those in the control group, respectively. Neither the dopamine nor glutamate level was affected by 6-[2-(4 imidazolyl)ethylamino]-N-(trifluoromethylphenyl)heptanecarboxamide (HTMT), an H(1) agonist (100 nmol, i.c.v.). However, dimaprit, an H(2) agonist (100 nmol, i.c.v.) suppressed the peak values to 42% and 32%, respectively. Most neurons were degenerated 7 days after ischemia in control animals. Histologic outcome was alleviated by either histamine or dimaprit treatment, whereas HTMT did not affect the outcome. Although postischemic administration of mepyramine, an H(1) antagonist (5 nmol, i.c.v.), did not affect the histologic alleviation caused by preischemic treatment with histamine, ranitidine, an H(2) antagonist (30 nmol, i.c.v.), partly abolished the improvement caused by histamine. These results suggest that suppression of ischemic release of excitatory neurotransmitters by histamine H(2) action is a contributing factor in alleviation of histologic outcome. PMID- 14744601 TI - Lactoferrin enhances peripheral opioid-mediated antinociception via nitric oxide in rats. AB - Lactoferrin (LF) is a multifunctional protein found in various biological fluids. However, the peripheral action of lactoferrin remains unknown. In this study, peripherally applied bovine lactoferrin showed antinociceptive effect that was reversed by a mu-opioid receptor antagonist, D-Phe-Cys-Tyr-D-Trp-Orn-Thr-NH(2) (CTOP), or by a nitric oxide synthase (NOS) inhibitor, N(G)-nitro-L-arginine methyl ester (L-NAME), but not by an inactive enantiomer of L-NAME, N(G)-nitro-D arginine methyl ester (D-NAME), during phase 1 and phase 2 in the rat formalin test. Peripheral coadministration of a micro-opioid receptor agonist, morphine, with subeffective dose of bovine lactoferrin produced a potentiated antinociceptive effect compared to that of morphine alone during both phases in the formalin test. This potentiated antinociception by morphine with bovine lactoferrin was reversed by CTOP or by L-NAME. These results suggest that bovine lactoferrin exerts an antinociceptive activity via potentiation of the peripheral micro-opioidergic system, and that nitric oxide (NO) is involved in this potentiation. PMID- 14744602 TI - Nerve injury alters the effects of interleukin-6 on nociceptive transmission in peripheral afferents. AB - Interleukin-6 (IL-6) is markedly upregulated in the peripheral and central nervous systems following nerve injury; however, the functional effects of this are unclear. This study investigates the effect of peripheral interleukin-6 on nociceptive transmission in naive and neuropathic states. Using an in vitro rat skin-nerve preparation, 50 ng interleukin-6 inhibited responses of single nociceptive fibers to noxious heat. A 20-ng sample of interleukin-6 only inhibited heat responses in the presence of soluble interleukin-6 receptors. To examine in vivo effects of peripheral interleukin-6, extracellular recordings from dorsal horn neurons were made in anaesthetised naive, sham-operated and neuropathic (spinal nerve ligated) rats. Peripheral interleukin-6 (40-100 ng) markedly inhibited all naturally evoked neuronal responses in naive rats, yet only neuronal responses to heat in neuropathic rats. Behaviourally, intraplantar administration of interleukin-6 (0.01-1 microg) elicited ipsilateral thermal hypoalgesia in naive rats. Thus, interleukin-6 inhibits normal peripheral nociceptive transmission, yet such anti-nociceptive effects are attenuated following nerve injury in a modality-specific manner. PMID- 14744603 TI - Participation of the L-arginine-nitric oxide-cyclic GMP-ATP-sensitive K+ channel cascade in the antinociceptive effect of rofecoxib. AB - The antinociceptive effect of rofecoxib, a preferential inhibitor of cyclooxygenase-2, was assessed in the pain-induced functional impairment model in the rat. Systemic administration of rofecoxib generated a dose-dependent antinociceptive effect in rats injected with uric acid into the knee joint of the right hindlimb in order to produce nociception. Ipsilateral intra-articular pretreatment with N(G)-L-nitro-arginine methyl ester (L-NAME, an inhibitor of nitric oxide (NO) synthesis), 1H-(1,2,4)-oxadiazolo (4,2-a)quinoxalin-1-one (ODQ, an inhibitor soluble guanylyl cyclase), and the ATP-sensitive potassium channel blocker glibenclamide reversed the antinociceptive effect of rofecoxib p.o. However, ipsilateral intra-articular pretreatment with L-arginine (a NO substrate), or 3-morpholino-sydnonimine-HCl (SIN-1, a non-enzymatic donor of NO), potentiated the antinociceptive effect induced by rofecoxib. The present results suggest that, in addition to cyclooxygenase-2 inhibition, the antinociceptive effect of rofecoxib could also involve activation of the L-arginine-NO-cyclic GMP (cGMP) pathway, followed by opening of ATP-sensitive K+ channels at the peripheral level. PMID- 14744604 TI - Effect of K+ channel modulators on the antiallodynic effect of gabapentin. AB - The effect of K+ channel inhibitors on the antiallodynic activity induced by spinal gabapentin was assessed in rats. Ligation of L5 and L6 spinal nerves made the rats allodynic, whereas that intrathecal administration of gabapentin (25-200 microg) reduced tactile allodynia in a dose-dependent manner. Spinal pretreatment with glibenclamide (12.5-50 microg, ATP-sensitive K+ channel inhibitor), charybdotoxin (0.01-1 ng) or apamin (0.1-3 ng, large-and small-conductance Ca2+ activated K+ channel blockers, respectively), but not margatoxin (0.01-10 ng, voltage-dependent K+ channel inhibitor), significantly prevented gabapentin induced antiallodynia. Pinacidil (1-30 microg, K+ channel opener) significantly reduced nerve ligation-induced allodynia. Intrathecal glibenclamide (50 microg), charybdotoxin (1 ng) and apamin (3 ng), but not margatoxin (10 ng), significantly reduced pinacidil-induced antiallodynia. K+ channel inhibitors alone did not modify allodynia produced by spinal nerve ligation. Results suggest that gabapentin and pinacidil may activate Ca2+-activated and ATP-sensitive K+ channels in order to produce part of its spinal antiallodynic effect in the Chung model. PMID- 14744605 TI - Sibutramine induces potential-dependent exocytotic release but not carrier mediated release of dopamine and 5-hydroxytryptamine. AB - In order to clarify the mechanism underlying the anti-obesity effects of sibutramine, we examined the effects of sibutramine on extracellular levels of dopamine and 5-hydroxytryptamine (5-HT) through microdialysis in the striatum in unanesthetized and freely moving rats. Sibutramine (5 mg/kg, oral administration (p.o.)) increased extracellular dopamine and 5-HT levels in rat striatum. The tricyclic antidepressant dosulepin (80 mg/kg, p.o. or 1 microM perfusion through the striatal probe) increased 5-HT levels only. Sibutramine-induced dopamine release was antagonized by perfusion of tetrodotoxin (1 microM) through the microdialysis probe in the striatum. However, sibutramine-induced dopamine release was not inhibited by prazosin (1 mg/kg, intraperitoneal injection (i.p.)), a suppressor of serotonergic activity in the striatum via blockade of alpha(1)-adrenoceptors, or perfusion with nomifensine (1 microM), an inhibitor of dopamine re-uptake. These results suggest that sibutramine increases dopamine levels in the striatum by exocytotic release and not by a carrier-mediated mechanism. PMID- 14744606 TI - Decrease in serotonin concentration in raphe magnus nucleus and attenuation of morphine analgesia in two mice models of neuropathic pain. AB - The alleviation of neuropathic pain cannot be satisfactorily achieved by treatment with opioids. There is much evidence to indicate that the active site of morphine for inducing effective analgesia is in the raphe magnus nucleus, where serotonin (5-HT, 5-hydroxytryptamine) acts as a primary transmitter. Therefore, we developed the hypothesis that 5-HT released in the raphe magnus nucleus could be related to the effectiveness of morphine in two mice models of neuropathic pain, diabetic (DM)-induced neuropathy and sciatic nerve ligation (SL). Two weeks after a single administration of streptozotocin, or 10 days after sciatic nerve ligation, mice were subcutaneously (s.c.) injected with morphine at 3, 5 and 10 mg/kg. The antinociceptive effect of morphine was estimated in the tail-pinch test; 5-HT content was measured after induction of neuropathic pain by microdialysis followed by high-performance liquid chromatography with electrochemical detection (HPLC-ECD). Morphine produced as insufficient antinociceptive effect in SL mice at all doses compared with that in sham operated mice, while in DM mice, morphine given s.c. at 5 and 10 mg/kg produced antinociceptive effects compared with those in non-diabetic mice, but not at 3 mg/kg. The 5-HT content of dialysates, expressed as AUC for 75 min, in SL and DM mice was less than that in control mice. However, morphine given s.c. at 5 mg/kg did not significantly affect 5-HT levels in both mice models compared to their controls. These results suggest that the decrease in 5-HT levels in the raphe magnus nucleus may be related to attenuation of the analgesic effect of morphine caused by the abnormal pain state found in diabetes and partial peripheral nerve injury. PMID- 14744607 TI - Behavioural and neurochemical effects of phosphatidylserine in MPTP lesion of the substantia nigra of rats. AB - The present study investigated the effects of intranigral MPTP (1-methyl-4-phenyl 1,2,3,6-tetrahydropyridine) infusion on rats treated with phosphatidylserine and evaluated in two memory tasks and on striatal dopamine levels. The results indicated that MPTP produced a significant decrease in the avoidance number in comparison to sham-operated and non-operated rats submitted to a two-way avoidance task. MPTP-lesioned rats exhibited an increase in the latencies to find the platform in cued version of the water maze in comparison to sham-operated and non-operated animals. The tested toxin reduced striatal dopamine levels in comparison to sham-operated and non-operated groups. A final surprising result was that phosphatidylserine was unable to reverse the cognitive deficits produced by MPTP or the reduction of striatal dopamine levels. In conclusion, the data suggest that MPTP is a good model to study the early impairment associated with Parkinson's disease and phosphatidylserine did not improve the memory impairment induced by MPTP. PMID- 14744608 TI - Cholecystokinin fails to block the spinal inhibitory effects of nociceptin in sham operated and neuropathic rats. AB - Cholecystokinin (CCK) has a number of roles in the central nervous system and can reduce the analgesic effect of activation of mu (micro), delta (delta) and kappa (kappa) opioid receptors. CCK has been proposed to be a major reason for reduced effects of morphine after nerve injury. This study examines if CCK modulates the effect of the Opioid Receptor Like-1 (ORL1) agonist, nociceptin on dorsal horn neurone activity in vivo in the spinal nerve ligation model of neuropathic pain compared with sham-operated and naive rats. In naive and neuropathic rats nociceptin alone inhibited the C-fibre evoked response, post-discharge, wind-up and input, while in sham operated rats nociceptin did not cause any inhibition but by contrast caused a facilitation of post-discharge and wind-up. CCK alone had no significant effect, although did cause slight facilitation in the three groups. In the presence of CCK the inhibitory effect of nocieceptin was blocked in naive animals, but in contrast the inhibitory effect of nociceptin was enhanced by CCK in sham and neuropathic rats. These results emphasize the differences between ORL1 and other opioid receptors. This loss of the inhibitory effect of CCK on nociceptin after nerve injury may be of clinical interest in the treatment of neuropathic pain. PMID- 14744609 TI - Ethanol-induced increases in neuroactive steroids in the rat brain and plasma are absent in adrenalectomized and gonadectomized rats. AB - Peripheral administration of alcohol has been demonstrated to cause significant increases in neurosteroid levels in the brain and periphery. These findings have led to several theories suggesting a role for neurosteroids in the actions of alcohol. However, the anatomical sources of these steroids (e.g., brain or periphery) are as yet unknown. This study utilized gas chromatography/mass spectrometry (GC/MS) to assess the levels of several neuroactive steroids in plasma and brain frontal cortex 30-360 min following acute administration of alcohol (2 g/kg, i.p.). Concentrations of pregnenolone, allopregnanolone (3alpha hydroxy-5alpha-pregnan-20-one), and allotetrahydrodeoxycorticosterone (3alpha,21 dihydroxy-5alpha-pregnan-20-one) were all measured. In order to determine the contribution of peripheral endocrine organs to neurosteroid responses, neuroactive steroid levels were measured in both intact and adrenalectomized/gonadectomized male Wistar rats 30 min after acute administration of alcohol. Intact animals exhibited a maximal increase of pregnenolone in plasma and frontal cortex 30 min after acute administration of alcohol. In addition, allopregnanolone levels increased, with a maximal effect observed at 60 min in plasma. However, in the adrenalectomized/gonadectomized groups treated with alcohol, no significant increases of pregnenolone, allopregnanolone, or allotetrahydrodeoxycorticosterone were found after 30 min. Thus, the alcohol-induced response was associated first with a relatively rapid increase in the first and rate-limiting step in the conversion of cholesterol to steroids, leading to increases in pregnenolone levels. This response was followed by the further secretion of the anxiolytic neuroactive steroids allopregnanolone and allotetrahydrodeoxycorticosterone, both of which appeared to be of adrenal and gonadal origin. PMID- 14744610 TI - In vivo pharmacological actions of two novel inhibitors of anandamide cellular uptake. AB - Two inhibitors of the cellular uptake of the endocannabinoid anandamide, (R)-N oleoyl-(1'-hydroxybenzyl)-2'-ethanolamine and (S)-N-oleoyl-(1'-hydroxybenzyl)-2' ethanolamine (OMDM-1 and OMDM-2, respectively), were recently synthesized, and their in vitro pharmacological activity described. Here we have assessed their activity in two typical pharmacological responses of cannabimimetic compounds. We first examined whether these compounds exert any effect per se on locomotion and pain perception in rats, and/or enhance the effects of anandamide on these two processes. We compared the effects of the novel compounds with those produced by a previously developed selective inhibitor, N-arachidonoyl-(2-methyl-4 hydroxyphenyl)amine (VDM-11). When assayed alone, OMDM-1 and OMDM-2 (1-10 mg/kg, i.p.) did not affect any of the five motor parameters under investigation, although the former compound exhibited a trend for the inhibition of ambulation, fast movements, and speed in rats. OMDM-2 and, to a lesser extent, VDM-11 (5 mg/kg, i.p.) enhanced the motor-inhibitory effects of a noneffective dose (2 mg/kg, i.p.) of anandamide, while OMDM-1 did not. In a typical test of acute analgesia, OMDM-2 and VDM-11 (1-10 mg/kg, i.p.), but not OMDM-1, significantly enhanced the time spent by rats on a "hot plate." However, the same compounds (5 mg/kg, i.p.) did not enhance the analgesic effect of a subeffective dose (2 mg/kg, i.p.) of anandamide, whereas OMDM-1 exerted a strong trend towards potentiation (P=0.06). We next explored the possible use of the two novel compounds in a pathological condition. Thus, we determined if, like other previously developed anandamide reuptake inhibitors, OMDM-1 and OMDM-2 inhibit spasticity in an animal model of multiple sclerosis-the chronic relapsing experimental allergic encephalomyelitis in mice. As previously shown with a higher dose of VDM-11, both novel compounds (5 mg/kg, i.v.) significantly reduced spasticity of the hindlimb in mice with chronic relapsing experimental allergic encephalomyelitis. We suggest that OMDM-1 and, particularly, OMDM-2 are useful pharmacological tools for the study of the (patho)physiological role of the anandamide cellular uptake process, and represent unique templates for the development of new antispastic drugs. PMID- 14744611 TI - Chronic 1,4-butanediol treatment in rats: cross-tolerance to gamma hydroxybutyrate and (+/-)-baclofen. AB - The effects of 1,4-butanediol, gamma-hydroxybutyrate (GHB), and (+/-)-baclofen on food-maintained responding in rats were assessed before, during, and after chronic treatment with 1,4-butanediol. Six weeks of treatment with 1,4-butanediol (twice daily, 320 mg/kg for 3 weeks followed by 560 mg/kg for 3 weeks) decreased sensitivity to the rate-decreasing effects of (+/-)-baclofen and GHB without changing sensitivity to 1,4-butanediol. Sensitivity to (+/-)-baclofen and GHB returned to control values 2-3 weeks after discontinuation of treatment. These data suggest that tolerance to the effects of GHB or its precursors might result from changes in GABA(B) mechanisms. PMID- 14744612 TI - Increased vasoconstriction to noradrenaline by 1400W, inhibitor of iNOS, in rats with streptozotocin-induced diabetes. AB - There is evidence that inducible nitric oxide synthase (iNOS) is activated at the acute phase of diabetes. We examined if selective inhibition of iNOS by 1400W (N 3-aminomethyl-benzyl-acetamidine) increases vascular response to noradrenaline in rats with streptozotocin (60 mg/kg i.v.)-induced diabetes for a duration of 3 weeks. The effects of noradrenaline on mean arterial pressure (MAP; 6, 16, 45 and 122x10(-9) mol/kg/min) and mean circulatory filling pressure (MCFP; 16 and 45x10( 9) mol/kg/min) were obtained in conscious and unrestrained diabetic rats and control rats before as well as after treatment with 1400W (3 mg/kg followed by 3 mg/kg/h, i.v.). Rats with early streptozotocin-induced diabetes had decreased mean arterial pressure and mean circulatory filling pressure responses to noradrenaline. Treatment with 1400W did not affect responses in the control rats but increased maximum pressor response to noradrenaline (from 46+/-3 to 63+/-5) and mean circulatory filling pressure response to the high dose (45 nmol/kg/min) of noradrenaline (from 1.0+/-0.2 to 3.8+/-0.3 mmHg) in the diabetic rats. Thus, selective inhibition of iNOS by 1400W increases arterial and venous constriction to noradrenaline in conscious rats with streptozotocin-induced diabetes. PMID- 14744613 TI - Lack of adenosine A1 and dopamine D2 receptor-mediated modulation of the cardiovascular effects of the adenosine A2A receptor agonist CGS 21680. AB - Some behavioral and biochemical effects of the systemically administered adenosine A(2A) receptor agonist 2-p-(2-carboxyethyl)phenethylamino-5'-N ethylcarboxamidoadenosine (CGS 21680) in rats are potentiated by adenosine A(1) receptor agonists and counteracted by dopamine D2 receptor agonists. In the present study we compared potentiating and antagonistic interactions between CGS 21680 and adenosine A(1) and dopamine D2 receptor agonists on motor activity and on cardiovascular responses (arterial blood pressure and heart rate). The motor depressant effects produced by CGS 21680 (0.5 mg/kg, i.p.) were potentiated by the adenosine A(1) receptor agonist N(6)-cyclopentyladenosine (CPA, 0.3 mg/kg, i.p.) and counteracted by the dopamine D2 receptor agonist quinpirole (0.5 mg/kg, i.p.). In contrast, neither CPA nor quinpirole significantly modified the decrease in arterial pressure or the increase in heart rate induced by CGS 21680. However, the adenosine A(2A) receptor antagonist 3-(3-hydroxypropyl)-8-(m methoxystyryl)-7-methyl-1-propargylxanthine phosphate disodium salt (MSX-3, 3 mg/kg, i.p.) counteracted both the motor-depressant and cardiovascular effects of CGS 21680. Therefore, the effects of the systemically administered adenosine A(2A) receptor agonist CGS 21680 on cardiovascular function, in contrast to its effects on motor behavior, appear to be independent of the effects of adenosine A(1) and dopamine D2 receptor activity. PMID- 14744614 TI - Endothelium-dependent noradrenergic hyperresponsiveness induced by thapsigargin in human saphenous veins: role of thromboxane and calcium. AB - To further investigate the mechanisms which regulate sympathetic vascular tone, we studied the effects of the sarcoplasmic reticulum Ca(2+)-ATPase inhibitor, thapsigargin, on the vasoconstriction induced by transmural nerve stimulation and noradrenaline in superfused human saphenous vein rings. The contractions induced by both transmural nerve stimulation and noradrenaline were potentiated by thapsigargin in endothelium-intact, but not in endothelium-denuded vessels. This potentiation was unaffected by the non-selective endothelin ET(A/B) receptor antagonist, Ro 47-0203 (4-tert-Butyyl-N-[6-(2-hydroxy-ethoxy)-5-(2-methoxy phenoxy)-2,2'-bipyrimidin-4yl]benzene sulfonamide), or by the nitric oxide (NO) synthase inhibitor, L-NNA (N(omega)-nitro-L-arginine), but was inhibited by the thromboxane A(2) receptor antagonist, Bay u3405 (3(R)-[[(4-flurophenyl) sulphonyl]amino-1,2,3,4-tetrahydro-9H-carbazole-9-propanoic acid]) or by the thromboxane A(2) synthase inhibitor, UK 38485 (3-(1H-imidazol-1-yl-methyl)-2 methyl-1H-indole-1-propanoic acid). Moreover, the thapsigargin-induced noradrenergic hyperresponsiveness, as well as that produced by subthreshold concentrations of the thromboxane A(2) mimetic, U 46619, were blocked by the Ca(2+) channel antagonist, verapamil. In conclusion, our results indicate that thapsigargin enhances the contractions produced by sympathetic nerve stimulation in human saphenous vein rings through the endothelial release of thromboxane A(2) that potentiates the vasoconstriction induced by the noradrenergic mediator with a verapamil-sensitive mechanism. PMID- 14744615 TI - 5-HT1B receptors and alpha 2A/2C-adrenoceptors mediate external carotid vasoconstriction to dihydroergotamine. AB - Dihydroergotamine produces external carotid vasoconstriction in vagosympathectomized dogs by 5-HT(1B/1D) receptors and alpha(2)-adrenoceptors. This study identified the specific subtypes involved in this response. One-minute intracarotid infusions of dihydroergotamine (5.6-10 microg/min) dose-dependently decreased external carotid blood flow without affecting blood pressure or heart rate. This response was: (1) partly blocked in dogs pretreated intravenously with the antagonists SB224289 (5-HT(1B); 2,3,6,7-tetrahydro-1'-methyl-5-[2'-methyl-4' (5-methyl-1,2,4-oxadiazol-3-yl)biphenyl-4-carbonyl]furo[2,3-f]indole-3-spiro-4' piperidine hydrochloride), rauwolscine (alpha(2)), BRL44408 (alpha(2A); 2-[2H-(1 methyl-1,3-dihydroisoindole)methyl]-4,5-dihydroimidazole) or MK912 (alpha(2C); (2S,12bS)-1'3'-dimethylspiro(1,3,4,5',6,6',7,12b-octahydro-2Hbenzo[b]furo[2,3 a]quinazoline)-2,4'-pyrimidin-2'-one); (2) markedly blocked after SB224289 plus rauwolscine; and (3) unaffected after BRL15572 (5-HT(1D); 1-(3-chlorophenyl)-4 [3,3-diphenyl (2-(S,R) hydroxypropanyl) piperazine] hydrochloride) or imiloxan (alpha(2B)). Therefore, the above response involves 5-HT(1B) receptors and alpha(2A/2C)-adrenoceptors. PMID- 14744616 TI - The action of extracellular NAD+ on Ca2+ efflux, hemodynamics and some metabolic parameters in the isolated perfused rat liver. AB - The action of NAD+ on hemodynamics and metabolism of the isolated perfused rat liver was investigated. Extracellular NAD+ (20-100 microM) stimulated glycogen breakdown (glucose release) and inhibited oxygen uptake. Lactate production was predominantly increased, and pyruvate production was predominantly inhibited. NAD+ also increased the portal perfusion pressure. All metabolic effects were strictly Ca2+-dependent. The effects were absent when Ca2+ was excluded, and reintroduction of the cation restored the effects. In preloaded livers, NAD+ accelerated 45Ca2+ efflux. The action of NAD+ was sensitive to three inhibitors of eicosanoid synthesis, suggesting that this action is mediated by these compounds, which are known to be produced and released by Kupffer and endothelial cells. It is impossible to infer from the available data if NAD+ exerts all these effects by itself or if they are caused by one or more of its extracellular hydrolysis products. Nicotinamide was ineffective and can be excluded, but especially cyclic ADP-ribose and ADP-ribose are possibilities that should be considered in future work. PMID- 14744617 TI - Use of radiotelemetry to evaluate respiratory depression produced by chronic methadone administration. AB - Illicit and therapeutic opioid administration can result in overdose due to opioid-induced respiratory depression. Research investigating the respiratory depressant effects of opioids has been limited due to difficulties associated with acquiring long-term respiratory data. This study examined the novel use of radiotelemetry to measure respiratory rate, heart rate, locomotor activity and blood pressure in rats treated chronically with methadone. Over 4 days of treatment, respiratory rate decreased, but partial tolerance appeared to develop during active (night) periods. Decreased heart rate was observed during the night periods and tolerance appeared to develop to this effect. Activity and blood pressure did not change with treatment. The effects of naloxone hydrochloride and naloxone methiodide administration on the methadone-treated rats were also examined and both antagonists increased respiratory rate and heart rate, with only naloxone hydrochloride producing significant increases in activity. Radiotelemetry offers a means of evaluating drug effects on respiratory rate continually in ambulatory, unstressed animals. PMID- 14744618 TI - Antiemetic effect of dexamethasone on cisplatin-induced early and delayed emesis in the pigeon. AB - We investigated the ability of dexamethasone to attenuate cisplatin (4 mg/kg, i.v.)-induced early and delayed emesis. These appear within the first 8-h period (early phase) and between 8 and 48 h (delayed phase), respectively, after cisplatin administration in the pigeon. Dexamethasone (0.1 and 1 mg/kg, i.m.) reduced significantly the number of emetic responses to cisplatin by 56% and 82% (P<0.05), respectively, in the early phase, and by 41% and 66% (P<0.05), respectively, in the delayed phase. Dexamethasone (1 and 10 microg/kg, i.c.v.) reduced the number of emetic responses by 66% and 91% (P<0.05), respectively, in the early phase, and by 56% and 87% (P<0.05), respectively, in the delayed phase. Indomethacin (10 mg/kg, i.m.) did not suppress cisplatin-induced early and delayed emesis. Dexamethasone (1 mg/kg, i.m.) did not affect the content of platinum in the medulla oblongata after cisplatin administration. The above results suggest that dexamethasone has antiemetic effects on both the early and delayed emetic responses to cisplatin in pigeons, partially via its central site of action, and that the antiemetic mechanism of dexamethasone is related to factors other than its inhibition of prostanoid synthesis or its membrane stabilizing effect which reduces influx of cisplatin into the medulla oblongata. PMID- 14744619 TI - Stereoselectivity for interactions of agonists and antagonists at mouse, rat and human beta3-adrenoceptors. AB - This study examines the stereoselectivity profile of recombinant mouse, rat and human beta(3)-adrenoceptors expressed in Chinese Hamster Ovary (CHO-K1) cells using radioligand binding, in comparison with endogenously expressed beta(3) adrenoceptors mediating relaxation responses in mouse ileum. The enantiomeric ratios for several beta-adrenoceptor agonists and antagonists at the cloned mouse, rat and human beta(3)-adrenoceptor were less than those reported at the cloned beta(1)-/beta(2)-adrenoceptor but higher than those reported in previous studies. The degree of stereoselectivity was relatively low for the agonists isoprenaline and noradrenaline but higher for antagonists and, in particular, tertatolol and propranolol. In mouse ileum, stereoselectivity of propranolol and tertatolol was observed under beta(1)-/beta(2)-adrenoceptor blockade. The (-) enantiomers of propranolol and tertatolol were more effective at antagonism of ( )-isoprenaline-mediated relaxation of mouse ileum than their (+)-enantiomers. The recombinant mouse, rat and human beta(3)-adrenoceptors display stereoselective interactions for agonists and antagonists similar to the stereoselective profile of beta(3)-adrenoceptors in mouse ileum. The degree of stereoselectivity varied between species and the human beta(3)-adrenoceptor displayed higher affinities and enantiomeric ratios than the mouse or rat beta(3)-adrenoceptors. PMID- 14744620 TI - Azithromycin reverses anticancer drug resistance and modifies hepatobiliary excretion of doxorubicin in rats. AB - The present study aims to investigate whether azithromycin reverses P glycoprotein-dependent anticancer drug resistance in vitro and modifies the hepatobiliary excretion of doxorubicin, a substrate for P-glycoprotein in vivo. Azithromycin increased dose-dependently the intracellular accumulation of doxorubicin in adriamycin-resistant human myelogenous leukemia cells (K562/ADR) with no effect on the expression of P-glycoprotein in the cells. However, the inhibitory effect was much weaker than that of cyclosporin A and was comparable to that of erythromycin. When Sprague-Dawley (SD) rats, which have drug transporting P-glycoprotein and multidrug resistance-associated protein 2 (Mrp2) in the bile canalicular membrane of hepatocytes, received an infusion of doxorubicin, the steady-state biliary clearance of doxorubicin was significantly decreased for 40 min after a single intravenous injection of azithromycin. However, azithromycin did not increase the plasma concentration of doxorubicin. The biliary clearance of doxorubicin in Eisai hyperbilirubinemic rats (EHBRs), which have a hereditary deficiency in Mrp2, was significantly decreased compared with that in Sprague-Dawley rats, suggesting the involvement of Mrp2 in the biliary excretion of doxorubicin. The present findings suggest that azithromycin overcomes P-glycoprotein-dependent anticancer drug resistance of tumors by inhibiting the binding of doxorubicin to P-glycoprotein in K562/ADR cells and inhibits the hepatobiliary excretion of drugs that are substrates for P glycoprotein and Mrp2. PMID- 14744621 TI - Effect of methylguanidine in carrageenan-induced acute inflammation in the rats. AB - In vitro and in vivo studies have demonstrated that methylguanidine, an inhibitor of nitric oxide synthase (NOS), is also able to reduce tumour necrosis factor alpha (TNF-alpha) release. In the present study, we evaluated the anti inflammatory potential of methylguanidine treatment in two models of acute inflammation (carrageenan-induced paw edema and pleurisy) where oxyradical, nitric oxide (NO) and prostaglandins play a crucial role in the inflammatory processes. Our data show that methylguanidine, given intraperitoneally at the dose of 30 mg/kg, inhibits the inflammatory response reducing significantly (P<0.05) paw swelling, pleural exudates formation, mononuclear cell infiltration and histological injury. Furthermore, our data suggests that there is a significant (P<0.05) reduction in the activity and expression both of the inducible NOS (iNOS) and of cyclooxygenase-2 in lung tissue of pleurisy model. Methylguanidine is also able to reduce the appearance of nitrotyrosine and of the nuclear enzyme poly(adenosine diphosphate [ADP]-ribose) synthase immunoreactivity in the inflamed lung tissues. Treatment with aminoguanidine, the reference drug, significantly reduced all the evaluated pro-inflammatory parameters in carrageenan-treated rats. Taken together, the present results demonstrate that methylguanidine exerts potent anti-inflammatory effects that could be, in part, related to an inhibition of the expression/activity of the iNOS and cyclooxygenase-2 and, another part, may be related to a reduction of TNF-alpha release. PMID- 14744622 TI - Effect of the K+(ATP) channel opener, KCO912, on baseline and allergen induced airway hyperresponsiveness in allergic rabbits. AB - The effect of the adenosine triphosphate sensitive K+ (K(ATP)) channel opener (3S,4R)-3,4-dihydro-3-hydroxy-2,2-dimethyl-4-(2-oxo-1-piperidinyl)-N-phenyl-1 benzopyran-6-sulphonamide (KCO912) on airway hyperresponsiveness induced using either a combination of allergen immunization (i.p.) followed by aerosol allergen challenge or immunization alone was investigated. Rabbits were immunized with Alternaria tenuis for the first 3 months of life. Airway responsiveness to histamine was measured 24 h before and after A. tenuis aerosol challenge. Fifteen minutes before the second challenge, rabbits were pre-treated with 10 microg of KCO912 or vehicle by inhalation. Allergen challenge induced airway hyperresponsiveness in vehicle pre-treated rabbits and pre-treatment with KCO912 abolished the airway hyperresponsiveness. The effect of KCO912 (10 microg) or vehicle on baseline airway hyperresponsiveness to the adenosine A(1) receptor agonist, cyclopentyl adenosine (CPA), induced by immunization with A. tenuis alone, was also assessed. Rabbits, immunized with A. tenuis alone, exhibited baseline airway hyperresponsiveness as demonstrated by an increase in airway resistance to CPA. Treatment with KCO912 did not alter the allergen-induced airway responsiveness to CPA. This study demonstrates that KCO912 can inhibit allergen-induced exacerbations of airway hyperresponsiveness. PMID- 14744623 TI - Significance of chymase inhibition for prevention of adhesion formation. AB - To clarify the role of chymase in adhesion formation, we investigated whether a chymase inhibitor could prevent adhesion formation after surgery in hamsters. Hamsters received a lesion produced by uterus scraping. A specific chymase inhibitor, 2-[4-(5-fluoro-3-methylbenzo[b]thiophen-2-yl)sulfonamido-3 methanesulfonylphenyl]oxazole-4-carboxylicacid (TY-51184), or placebo was injected into the abdomen before closing and scores for adhesion formation were assessed at 1, 4, and 12 weeks. A single peritoneal administration of TY-51184 significantly decreased the adhesion scores even at 12 weeks (placebo, 2.80+/ 0.20; chymase inhibitor, 1.60+/-0.31). Thus, chymase inhibitors may be a novel strategy to prevent adhesion formation. PMID- 14744624 TI - Chronic vasopeptidase inhibition normalizes diabetic endothelial dysfunction. AB - Type 2 diabetes mellitus is a major cause of vascular morbidity but animal models for this disease have not been adequately characterized. We demonstrate that endothelial dysfunction is present in the Zucker diabetic fatty (ZDF) rat. Vasopeptidase inhibition with AVE7688 (7-[[(2S)-2-(acetylthio)-1-oxo-3 methylpropyl]amino]-1,2,3,4,6,7,8,12b-octahydro-6-oxo-(4S,7S,12bR)-pyrido[2,1 a][2]benzazepine-4-carboxic acid), 45 mg/kg/day in chow for 6 weeks, normalized acetylcholine mediated relaxation of mesenteric artery rings. Thus, chronic vasopeptidase inhibition may prevent vascular complications related to type 2 diabetes mellitus. PMID- 14744625 TI - Inhibition of basal and interleukin-1-induced VCAM-1 expression by phospholipid hydroperoxide glutathione peroxidase and 15-lipoxygenase in rabbit aortic smooth muscle cells. AB - Cytokines or hydroperoxides upregulate cell adhesion molecules (CAM) in early stages of atherosclerosis. VCAM-1 expression was therefore investigated in rabbit aortic smooth muscle cells (SMC) stably transfected either with phospholipid hydroperoxide glutathione peroxidase (PHGPx; SMCPHGPx) as a hydroperoxide reducing enzyme or with 15-lipoxygenase (15-LOX; SMCLOX) as a hydroperoxide producing enzyme. Transfected cells showed up to 3-fold enhanced PHGPx and a marked LOX activity, respectively, that was absent in controls. Intracellular hydroperoxides were 6-fold higher in SMCLOX than in SMC or SMCPHGPx. Intracellular protein thiols were decreased by 50 and 90% in SMCPHGPx and SMCLOX, respectively. Glutathione mixed disulfides were tentatively increased from SMC via SMCPHGPx to SMCLOX, accordingly. Thiol reduction with tris(2 carboxyethyl)phosphine completely restored protein thiols in SMCPHGPx, whereas in SMCLOX only 60% of control values were recovered. Basal VCAM-1 mRNA levels were decreased by 50% in SMCPHGPx and 75% in SMCLOX. VCAM-1-inducibility was abrogated in SMCLOX but not in SMCPHGPx. Accordingly, NFkappaB-driven reporter gene activation by IL-1 was unaffected in SMCPHGPx but abolished in SMCLOX. The data confirm that PHGPx overexpression dampens CAM expression either by lowering stimulatory hydroperoxides or by using hydroperoxides for protein modification. But hydroperoxides, when constitutively overproduced as in SMCLOX, inhibit CAM expression and render cells refractory to IL-1 stimulation likely due to oxidation of protein thiols of the signaling system. PMID- 14744626 TI - Variability of glutathione during the menstrual cycle-due to estrogen effects on hepatocytes? AB - Oxidative stress and alterations in the antioxidative defense system may be involved in carcinogenesis. We have previously shown that the levels of glutathione (GSH) in vivo in both breast tissue and subcutaneous fat were higher in the luteal phase compared with the follicular phase, suggesting an overall increase in GSH. This result was confirmed in the present study. Moreover, we exposed normal breast tissue in vivo, breast epithelial cells in vitro, and hepatocytes in culture to ovarian hormones. We found that local perfusion with estradiol, using microdialysis, in normal human breast tissue did not alter the local GSH levels in vivo. In vitro, treatment with estradiol and progesterone of normal human breast epithelial cells did not alter GSH levels. However, levels of GSH in hepatocytes were after 8 h estradiol exposure initially decreased, 76.6 +/ 5% of control cells, p <.05, whereas 20 h exposure more than doubled GSH, 209 +/ 26% compared with control cells, p <.01. Progesterone had no additional effect. Exposure of hepatocytes to estradiol increased the cellular content of gamma glutamylcysteine synthetase, the rate-limiting enzyme in GSH synthesis. In conclusion we suggest that estradiol affects the GSH homeostasis mainly by effects on hepatocytes, whereas local production in the breast is unaffected by estradiol. PMID- 14744627 TI - Peroxynitrite-mediated alpha-tocopherol oxidation in low-density lipoprotein: a mechanistic approach. AB - Previous reports proposed that peroxynitrite (ONOO-) oxidizes alpha-tocopherol (alpha-TOH) through a two-electron concerted mechanism. In contrast, ONOO- oxidizes phenols via free radicals arising from peroxo bond homolysis. To understand the kinetics and mechanism of alpha-TOH and gamma-tocopherol (gamma TOH) oxidation in low-density lipoprotein (LDL) (direct vs. radical), we exposed LDL to ONOO- added as a bolus or an infusion. Nitric oxide (.NO), ascorbate and CO2 were used as key biologically relevant modulators of ONOO- reactivity. Although approximately 80% alpha-TOH and gamma-TOH depletion occurred within 5 min of incubation of 0.8 microM LDL with a 60 microM bolus of ONOO-, an equimolar infusion of ONOO- over 60 min caused total consumption of both antioxidants. gamma-Tocopherol was preserved relative to alpha-TOH, probably due to gamma tocopheroxyl radical recycling by alpha-TOH. alpha-TOH oxidation in LDL was first order in ONOO- with approximately 12% of ONOO- maximally available. Physiological concentrations of.NO and ascorbate spared both alpha-TOH and gamma-TOH through independent and additive mechanisms. High concentrations of.NO and ascorbate abolished alpha-TOH and gamma-TOH oxidation. Nitric oxide protection was more efficient for alpha-TOH in LDL than for ascorbate in solution, evidencing the kinetically highly favored reaction of lipid peroxyl radicals with.NO than with alpha-TOH as assessed by computer-assisted simulations. In addition, CO2 (1.2 mM) inhibited both alpha-TOH and lipid oxidation. These results demonstrate that ONOO induces alpha-TOH oxidation in LDL through a one-electron free radical mechanism; thus the inhibitory actions of.NO and ascorbate may determine low alpha-tocopheryl quinone accumulation in tissues despite increased ONOO- generation. PMID- 14744628 TI - Isomer-specific contractile effects of a series of synthetic f2-isoprostanes on retinal and cerebral microvasculature. AB - F2-isoprostanes (F2-IsoP's) are biologically active prostanoids formed by free radical-mediated peroxidation of arachidonic acid. Four different F2-IsoP regioisomers (5-, 8-, 12-, and 15-series), each comprising eight racemic diastereomers, total 64 compounds. Information regarding the biological activity of IsoP's is largely limited to 15-F2t-IsoP (8-iso-PGF2alpha). We recently demonstrated that 15-F2t-IsoP and its metabolite, 2,3-dinor-5,6-dihydro-15-F2t IsoP, evoked vasoconstriction and TXA2 generation in retina and brain microvasculature. We have now examined and compared the biological activities of a series of recently synthesized new 5-, 12-, and 15-series F2-IsoP isomers in pig retinal and brain microvasculature. We hereby show that other 15-series F2 IsoP isomers, 15-epi-15-F2t-IsoP, ent-15-F2t-IsoP, and ent-15-epi-15-F2t-IsoP, are also potent vasoconstrictors. The 12-series isomers tested, 12-F2t-IsoP and 12-epi-12-F2t-IsoP, also caused marked vasoconstriction. Of the 5-series isomers tested, 5-F2t-IsoP and 5-epi-5-F2t-IsoP possessed no vasomotor properties, whereas ent-5-F2t-IsoP caused modest vasoconstriction. The vasoconstriction of ent-5-F2t-IsoP, 12-F2t-IsoP, and 12-epi-12-F2t-IsoP was abolished by removal of the endothelium, by TXA2 synthase and receptor inhibitor (CGS12970, L670,596), and by receptor-mediated Ca2+ channel blockade (SK & F96365); correspondingly, these isomers increased TXB2 formation by activating Ca2+ influx (detected with fura 2-AM) through non-voltage-dependent receptor-mediated Ca2+ entry (SK & F96365 sensitive) in endothelial cells. In conclusion, as seen with 15-F2t-IsoP, ent-5-F2t-IsoP, 12-F2t-IsoP, and 12-epi-12-F2t-IsoP constricted both retinal and brain microvessels by inducing endothelium-dependent TXA2 synthesis. These new findings broaden the scope of our understanding regarding the potential involvement of F2-IsoP's as mediators of oxidant injury. PMID- 14744629 TI - Collaborative effects of Photobacterium CuZn superoxide dismutase (SODs) and human AP endonuclease in DNA repair and SOD-deficient Escherichia coli under oxidative stress. AB - The defenses against free radical damage include specialized repair enzymes that correct oxidative damage in DNA and detoxification systems such as superoxide dismutases (SODs). These defenses may be coordinated genetically as global responses. We hypothesized that the expression of SOD and DNA repair genes would inhibit DNA damage under oxidative stress. Therefore, protection of Escherichia coli mutants deficient in SOD and DNA repair genes (sod-, xth-, and nfo-) was demonstrated by transforming the mutant strain with a plasmid pYK9 that encoded Photobacterium leiognathi CuZnSOD and human AP endonuclease. The results show that survival rates were increased in sod+ xth- nfo+ cells compared with sod- xth ape-, sod- xth- ape-, and sod+ xth- ape- cells under oxidative stress generated with 0.1 mM paraquat or 3 mM H2O2. The data suggest that, at the least, SOD and DNA repair enzymes may collaborate on protection and repair of damaged DNA. Additionally, both enzymes are required for protection against free radicals. PMID- 14744630 TI - Genistein ameliorates beta-amyloid peptide (25-35)-induced hippocampal neuronal apoptosis. AB - beta-Amyloid protein (Abeta), a major component of senile plaques of Alzheimer's disease (AD) brain, causes elevation of the intracellular free Ca2+ level and the production of robust free radicals, both of which contribute greatly to the AD associated cascade including severe neuronal loss in the hippocampus. Genistein, the most active molecule of soy isoflavones, protects diverse kinds of cells from damage caused by a variety of toxic stimuli. In the present study, we investigated the neuroprotective effect of genistein against Abeta25-35-induced apoptosis in cultured hippocampal neurons, as well as the underlying mechanism. Abeta25-35-induced apoptosis, characterized by decreased cell viability, neuronal DNA condensation, and fragmentation, is associated with an increase in intracellular free Ca2+ level, the accumulation of reactive oxygen species (ROS), and the activation of caspase-3. All these phenotypes induced by Abeta25-35 are reversed by genistein. Our results further show that at the nanomolar (100 nM) level, genistein protects neurons from Abeta25-35-induced damage largely via the estrogen receptor-mediated pathway, and at the micromolar (40 microM) level, the neuroprotective effect of genistein is mediated mainly by its antioxidative properties. Our data suggest that genistein attenuates neuronal apoptosis induced by Abeta25-35 via various mechanisms. PMID- 14744631 TI - Low-affinity receptor-mediated induction of superoxide by N-formyl-methionyl leucyl-phenylalanine and WKYMVm in IMR90 human fibroblasts. AB - We investigated in IMR90 cells the effects of N-formyl-Met-Leu-Phe (N-fMLP) and WKYMVm (W peptide) on activation of the NADPH oxidase-like enzyme. In serum deprived human fibroblasts, exposure to 100 microM N-fMLP or 10 microM peptide W for 1 min induced both p47phox translocation and NADPH-dependent superoxide generation. These effects were in large part mediated by prevention of the rapid activation of extracellular signal-regulated kinases (ERKs) by preincubation with the MEK1 inhibitor PD098059. Furthermore, responses to N-fMLP or W peptide were inhibited by pertussis toxin, suggesting the involvement of a seven-transmembrane G protein-coupled receptor(s) for peptides. RT-PCR experiments demonstrated the expression in these cells of the low-affinity receptor FPRL1, but not the high affinity receptor FPR. Incubation with radiolabeled WKYMVm, which had a higher efficiency on FPRL1, revealed that human fibroblasts express binding sites for 125I-WKYMVm that are specifically displaced by increasing concentrations of unlabeled ligand. Analysis of the binding data predicted a Kd of 155.99 nM and a receptor density of about 16,200 molecules/cell. HEK293 cells, which express a NADPH oxidase-like enzyme but not formyl peptide receptors, transiently transfected with FPRL1 cDNA produced superoxide on stimulation with N-fMLP or W peptide, demonstrating that this receptor is biologically functional. PMID- 14744632 TI - Relevance of apple polyphenols as antioxidants in human plasma: contrasting in vitro and in vivo effects. AB - Apples are a major source of flavonoids in the Western diet, and flavonoid-rich foods may help protect against chronic diseases by antioxidant mechanisms. In the present study we investigated: (1) the antioxidant capacity of representative apple polyphenols and their contribution to the total antioxidant capacity of apple extracts; (2) the effects of adding apple extract to human plasma in vitro on oxidation of endogenous antioxidants and lipids; and (3) the effects of apple consumption by humans on ex vivo oxidation of plasma antioxidants and lipids. We found that the apple-contained flavonols and flavanols, quercetin, rutin, (-) epicatechin, and (+)-catechin, had a higher antioxidant capacity than the dihydrochalcones, phloridzin and phloretin, and the hydroxycinnamate, chlorogenic acid. However, together these apple polyphenols contributed less than 20% to the total antioxidant capacity of aqueous apple extracts. When human plasma was exposed to a constant flux of aqueous peroxyl radicals, endogenous ascorbate (70.0 +/- 10.3 microM) was oxidized within 45 min of incubation, while endogenous urate (375 +/- 40 microM) and alpha-tocopherol (24.7 +/- 1.2 microM) were oxidized after ascorbate. Addition of 7.1 or 14.3 micrograms/ml total phenols of apple extract did not protect ascorbate from oxidation, but increased the half life (t1/2) of urate from 136 +/- 15 to 192 +/- 16 and 208 +/- 23 min, respectively (p < 0.05 each), and t1/2 of alpha-tocopherol from 141 +/- 18 to 164 +/- 8 min (p = ns) and 188 +/- 8 min (p < 0.05). Lipid peroxidation started after ascorbate depletion, and addition of apple extract increased the lag time preceding detectable lipid peroxidation from 36.3 +/- 3.7 to 50.9 +/- 2.7 min (p < 0.05) and 70.4 +/- 4.2 min (p < 0.001). However, when six healthy volunteers ate five apples and plasma was obtained up to 4 h after apple consumption, no significant increases in the resistance to oxidation of endogenous urate, alpha tocopherol, and lipids were found. Thus, despite the high antioxidant capacity of individual apple polyphenols and apple extracts and the significant antioxidant effects of apple extract added to human plasma in vitro, ingestion of large amounts of apples by humans does not appear to result in equivalent in vivo antioxidant effects of apple polyphenols. PMID- 14744633 TI - Colonic metabolism of dietary polyphenols: influence of structure on microbial fermentation products. AB - The metabolism of chlorogenic acid, naringin, and rutin, representative members of three common families of dietary polyphenols, the hydroxycinnamates, the flavanones, and the flavonols, respectively, was studied in an in vitro mixed culture model of the human colonic microflora. Time- and concentration-dependent degradation of all three compounds was observed, which was associated with the following metabolic events after cleavage of the ester or glycosidic bond: reduction of the aliphatic double bond of the resulting hydroxycinnamate caffeic acid residue; dehydroxylation and ring fission of the heterocyclic C-ring of the resulting deglycosylated flavanone, naringenin, and of the deglycosylated flavonol, quercetin (which differed depending on the substitution). The metabolic events, their sequences, and major phenolic end products, as identified by GC-MS or LC-MS/MS, were elucidated from the structural characteristics of the investigated compounds. The major phenolic end products identified were 3-(3 hydroxyphenyl)-propionic acid for chlorogenic acid, 3-(4-hydroxyphenyl)-propionic acid and 3-phenylpropionic acid for naringin, and 3-hydroxyphenylacetic acid and 3-(3-hydroxyphenyl)-propionic acid for rutin. The degree of degradation of the compounds studied was significantly influenced by the substrate concentration as well as individual variations in the composition of the fecal flora. The results support extensive metabolism of dietary polyphenols in the colon, depending on substrate concentration and residence time, with resultant formation of simple phenolics, which can be considered biomarkers of colonic metabolism if subsequently absorbed. It is also apparent that a relatively small number of phenolic degradation products are formed in the colon from the diverse group of natural polyphenols. PMID- 14744634 TI - Oxidative stress in human hypertension: association with antihypertensive treatment, gender, nutrition, and lifestyle. AB - There is growing evidence that oxidative stress contributes to the pathogenesis of hypertension. Our aim was to measure markers of oxidative stress in hypertensive subjects, and assess the potential confounding influences of antihypertensive therapy, other cardiovascular risk factors, gender, lifestyle, and nutrition. Markers of oxidative stress, including plasma and 24 h urinary F2 isoprostanes, were measured in 70 untreated (men = 43, women = 27) and 85 treated (men = 43, women = 42) hypertensive subjects and 40 normotensive controls (men = 20, women = 20). Overall, F2-isoprostanes were not elevated in hypertensive subjects compared with controls. However, urinary and plasma F2-isoprostanes were significantly lower in treated compared with untreated hypertensive men, but not women. In hypertensive men, the number of antihypertensive drugs taken was inversely associated with both urinary and plasma F2-isoprostanes (p <.05). Self reported alcohol intake and biomarkers of alcohol consumption (gamma-glutamyl transpeptidase and high-density lipoprotein cholesterol) were positively associated with plasma but not urinary, F2-isoprostanes in men. Several nutrients were independently associated with plasma and urinary F2-isoprostanes in women. The results do not support the hypothesis that treated or untreated hypertensive subjects are under increased oxidative stress compared with normotensive controls. PMID- 14744635 TI - Mechanistic analysis of the immunomodulatory effects of a catalytic antioxidant on antigen-presenting cells: implication for their use in targeting oxidation reduction reactions in innate immunity. AB - Reactive oxygen species (ROS) have an indispensable role in controlling the growth of pathogens. Recent evidence also suggests that they can function as second messengers and modulators of the immune system. The identification of many redox-sensitive signal transduction pathways that are necessary for initiating the innate proinflammatory immune response suggests that modulation of these oxidation-reduction reactions may provide a means of therapeutic benefit for controlling inflammatory-mediated diseases. In order to test this hypothesis we employed two catalytic antioxidants (AEOL 10113 and 10150) for the determination of the role of oxidation-reduction reactions in innate immune system activation. Catalytic antioxidants prevented the initiation of the innate immune response in LPS-stimulated macrophages as evidenced by the suppression of proinflammatory cytokines (TNF-alpha, IL-1beta) and ROS (NO2- and O2-). The suppression of proinflammatory cytokine and ROS production correlated with the inhibition of NF kappaB DNA binding, without any effects on the mitogen-activated protein kinase signaling pathway. Catalytic antioxidants prevented NF-kappaB from binding DNA by an oxidation mechanism that was reversible with the addition of DTT. Although the primary use of these agents was to reduce and scavenge ROS, surprisingly, we also observed the ability of these compounds to exhibit oxidoreductase activity and oxidize redox-sensitive transcription factors such as NF-kappaB. Catalytic antioxidants exhibit antioxidant and pro-oxidant activities and our data further demonstrate the importance of redox balance for the initiation of proinflammation. The coupling of the innate with the adaptive immune response is dependent on TNF-alpha, IL-1beta, NO2-, and O2- generation; therefore, agents like catalytic antioxidants that decrease proinflammatory cytokines and ROS may provide protective effects in diseases in which chronic inflammation plays a pathogenic role. PMID- 14744636 TI - Redox-sensitive mechanism of no scavenging by nitronyl nitroxides. AB - Nitronyl nitroxides, NN., have been increasingly used in the field of NO-related studies as specific antagonists of NO. . We employed a combination of EPR and NMR spin trapping to study the mechanisms of the reaction of NN. with NO. in reducing environments. EPR allowed observation of NO-induced transformation of the paramagnetic trap, NN., to the corresponding iminonitroxide, IN. . In a complementary way, corresponding EPR-invisible diamagnetic products (the hydroxylamines NN-H and IN-H) were detected by 19F-NMR using newly synthesized fluorinated traps. Addition of reducing agents to a solution of NN. resulted in fast disappearance of its EPR spectra and appearance of a 19F-NMR peak of the corresponding hydroxylamine, NN-H. Addition of NO. as a bolus, or NO. generated on sodium nitroprusside photolysis, resulted in 19F-NMR-detectable accumulation of the hydroxylamine, IN-H. Upon high rates of NO. generation in ascorbate containing solutions, partial recovery of NN. was observed, which undergoes further reactions with NO. and ascorbate in a competitive manner. Using 19F-NMR and a fluorinated trap, NO-induced conversion of NN-H into IN-H was also observed in vivo in hypertensive ISIAH rats compared with normotensive WAG rats. The results provide insight into a new potential redox-sensitive mechanism of the antagonistic action of NN. against NO., which may provide insight into previously unexplained behavior of this category of NO-reacting compounds. PMID- 14744638 TI - [Is aspirin resistance a reality?]. PMID- 14744637 TI - Selenomethionine protects against adverse biological effects induced by space radiation. AB - Ionizing radiation-induced adverse biological effects impose serious challenges to astronauts during extended space travel. Of particular concern is the radiation from highly energetic, heavy, charged particles known as HZE particles. The objective of the present study was to characterize HZE particle radiation induced adverse biological effects and evaluate the effect of D-selenomethionine (SeM) on the HZE particle radiation-induced adverse biological effects. The results showed that HZE particle radiation can increase oxidative stress, cytotoxicity, and cell transformation in vitro, and decrease the total antioxidant status in irradiated Sprague-Dawley rats. These adverse biological effects were all preventable by treatment with SeM, suggesting that SeM is potentially useful as a countermeasure against space radiation-induced adverse effects. Treatment with SeM was shown to enhance ATR and CHK2 gene expression in cultured human thyroid epithelial cells. As ionizing radiation is known to result in DNA damage and both ATR and CHK2 gene products are involved in DNA damage, it is possible that SeM may prevent HZE particle radiation-induced adverse biological effects by enhancing the DNA repair machinery in irradiated cells. PMID- 14744640 TI - [Influence of hypergammaglobulinemia on antiphospholipid antibodies titres]. AB - PURPOSE: Antiphospholipid antibodies (aPL), anticardiolipin antibodies (aCL) or lupus anticoagulant (LA), are indispensable for the diagnosis of antiphospholipid syndrome (APS). However, antiphospholipid assays can generate false positive results. MATERIALS: We have studied the influence of hypergammaglobulinemia (HG) on aPL antibodies titers in 232 patients twice as positive for aPL antibodies. RESULTS: Out of 232 patients, 93 have an APS (76 primary APS, 17 secondary APS). Thrombosis occurred 138 times in APS patients. Of 139 patients without APS, 95 have an auto-immune disease, 28 have an isolated prolonged KCT and 16 an evolutive neoplasia. LA seems to be the best marker of APS. On the other hand aCL IgG and M, anti-beta2-GP1 IgM titers are significantly higher in patients without APS but with HG. CONCLUSION: Those results suggest that biological APS diagnosis should be carefully performed in patients with HG. In this case, other additional risk factors must be considered for the etiological diagnosis of thrombosis. PMID- 14744639 TI - [Gaucher's disease in Tunisia (multicenter study)]. AB - Gaucher's disease is one of the rare lysosomial disease that could receive substitutive enzymatic treatment which may improve considerably the prognosis of certain forms. The purpose of this work is to study the epidemiology of the disease in Tunisia, to highlight the diagnostic and therapeutic difficulties and also to precise our subsequent needs for substitutive medication. PATIENTS AND METHODS: We have conducted a retrospective survey of the hospital wards that were susceptible to take care of patients having Gaucher's disease. These wards are the paediatric, neonatology, internal medicine, haematology, neurology and cardiology wards. RESULTS: In this study we have observed 27 cases of Gaucher's disease over a period of 18 years (1983-2001). The age at onset ranges from birth to 73 years of age, with an average age of 14.5 years. According to the age at onset and the clinical presentation, we classify our patients into: 20 cases of type 1 (74%), three cases of type 2 (12%), and three cases of type 3 (12%), and one case of unspecified type.Gaucher's disease type 1: The age at onset ranged from 10 months to 73 years with an average of 19 years. The main clinical signs that we have observed were splenomegaly, hepatomegaly, pallor, haemorrhagic appearance and also osteoporosis and bone pain observed in 40% of the cases. The diagnosis was based on histology showing the Gaucher's cells in various tissues while the diagnosis obtained by the dosage of glucocerebrosidase took place only in 50% of the cases. The treatment has always been symptomatic (analgesics, transfusion). A splenectomy was performed in 47% of the cases and none of the patients received a specific treatment. The follow-up period ranged from 1 month to 18 years with an average follow-up of 4 years. Among the 12 patients having a follow-up of at least 1 year, we have noticed an improvement after splenectomy in three cases, a stability in three cases and two worsening cases dealing mainly with bone problems. One patient aged 73 died from respiratory problem and three were lost to follow-up. Gaucher's disease type 2: We have observed three cases of Gaucher's disease type 2 diagnosed at 1 day, 45 days and 3 months of age. The visceral manifestations were serious and the neurological features included seizures, hypertony, ocular-nerve palsies and psychometric decline. The three patients died. Gaucher's disease type 3: Three patients were probably suffering from Gaucher's disease type 3 with visceral manifestations observed at the ages of 9 months, 1 year and 3 years, and also neurological signs observed at respective ages of 2.5 and 3 years. Two patients died and the remaining one was lost to follow-up. CONCLUSION: Gaucher's disease is not exceptional in Tunisia. Type 1 is by far the most common one. We have noticed some insufficiency in the diagnosis as the glucocerebrosidase enzymatic dosage was performed only in 50% of the cases as well as therapeutic insufficiency with no prescription of the specific treatment. PMID- 14744641 TI - [Cardiac troponin I and T: specific biomarkers of cardiomyocyte]. AB - PURPOSE: Cardiac troponin I and troponin T have replaced creatine kinase MB (CK MB) for the diagnosis of cardiomyocyte necrosis. Cardiac specificity of these new markers leads to a change in our practice. CURRENT KNOWLEDGE AND KEY POINTS: Following necrosis, intracellular proteins are released into blood. This easy concept overlaps a biological complexity since troponins are released as complexes leading to various cut-off values depending on the assay used, as least for cardiac troponin I. The increase in both specificity and analytical sensitivity of these markers reached to propose a new definition of myocardial infarction. The diagnosis of acute coronary syndrome is a clinical based diagnosis, the use of troponin contributing to their classification. Finally, pathological processes leading to cardiac injury may induce an increase in the cardiac troponin level. FUTURE PROSPECTS AND PROJECTS: Troponin standardization is a challenge for the near future leading to better follow-up of patients and comparison between cohorts. PMID- 14744642 TI - [Long term survival in metastatic colorectal cancer treated with leucovorin and 5 fluoro-uracil chemotherapy]. AB - PURPOSE: Median survival in advanced colorectal cancer patients treated with 5 fluoro-uracil (5FU) and leucovorin (LV) is between 12 and 18 months. The aim of this study was to evaluate long term survival in this disease. METHODS: We report here, retrospectively, the survival of 445 patients who entered in first-line prospective studies with LV-5FU-based regimen chemotherapy, between 1985 and 1995. RESULTS: Median survival was 18 months. The 3, 5 and 10 year survival were respectively 17.9%, 4.5% and 2.4%. Seventy-five patients survived more than 3 years, among them, 10 achieved a complete and 34 a partial response, 12 had curative liver or lung surgery. Fifteen patients lived more than 5 years, 2 achieved a complete and 7 a partial response. Seven had curative surgery. Eleven patients were still alive in 2002, among them 7 in complete remission at 5 years, including 3 who did not have surgery. CONCLUSION: This study shows that some patients with metastatic colorectal cancer can achieve long survival, especially when secondary curative surgery can be performed. However, 1% of the patients can be cured with LV-5FU chemotherapy alone. These results will be probably improve with the use of the new drugs: oxaliplatin and irinotecan. PMID- 14744643 TI - [AA Amyloidosis: recent knowledges on pathophysiology]. AB - PURPOSE: Amyloidosis is a rare disease associated with an underestimated frequency because of the need of a pathological diagnosis identifying extracellular deposits with affinity for Congo red. There are moreover 20 proteins that can form extracellular fibril deposits. Some amyloidosis forms are more common than others, especially AA amyloidosis and AL amyloidosis. Among genetic amyloidosis, the transthyretin related amyloidosis is the most prevalent. The amyloid frequency could also be increased if amyloidosis related to Alzheimer's disease or prion's disease is included. In the absence of specific treatment for amyloidosis, researches are focused on amyloidosis pathophysiology especially, on AA amyloid pathophysiology. CURRENT KNOWLEDGE AND KEY POINTS: Amyloid is not only composed of fibrils but also of proteoglycanes, P component and amyloid-enhancing factor. A new research aim is focused on the cells involved in amyloid formation and on the relationship between amyloid, proteoglycanes and P component. FUTURE PROSPECTS AND PROJECTS: It was demonstrated that, in the absence of macrophages, an extracellular amyloid formation was possible with amyloid-enhancing factor as starting point. Some inhibitors of intra or extracellular amyloid formation are still to be discovered. Anti-P component has been recently developed; it was successful in the treatment of murin AA amyloidosis and gave some hope concerning the treatment of human amyloidosis. PMID- 14744644 TI - [Conventional ultrasonography Doppler in the assessment of arterial peripheral circulation]. AB - Ultrasonography Doppler in the assessment of arterial peripheral circulation. PURPOSE: The ultrasonography-Doppler study of peripheral arterial circulation requires measurements of arterial area and arterial blood flow. The aim of this study was to assess their variabilities, intra- and interobserver. METHODS: We studied brachial arterial circulation of 13 volunteers. Two experimented investigators carried out the examinations. Arterial blood pressure measurements were obtained at the level of the brachial artery of the left arm by means of a semi automated oscillometric device. The brachial artery was approached by ultrasound scan in semi-decubitus position, the arm in exterior rotation and abduction. Brachial artery cross-sectional areas were obtained by a transversal view with a depth of 4-15-cm upstream the antecubital fossa of the forearm. Cross sectional areas were measured at end diastole (SD) and end systole (SS). We measured maximal systolic and diastolic velocities (Vmax and Vmin), mean velocity (Vmoy) and velocity integral (ITV) of the blood flow. We deduced brachial output (D), resistance and pulsatility index (IR, IP), cross-sectional compliance coefficient (C). RESULTS: Intra- and interoperator variability as assessed by the Bland-Altman approach was acceptable SS, SD, ITV, D, Vmax, Vmoy, Vmin C, IR, and IP, in regard with values of variation coefficient, biais, and concordance limits. CONCLUSION: Conventional B-mode ultrasonography may offer a reliable method for arterial peripheral circulation evaluation. PMID- 14744645 TI - [Involvement of thoracic aorta revealing temporal arteritis. Three cases]. AB - INTRODUCTION: Temporal arteritis involves large vessels in 15% of cases. Their discovery is usually late, commonly several years after the diagnosis. EXEGESIS: We describe three cases of temporal arteritis with thoracic aorta involvement: two patients presented with aorta aneurysm which revealed temporal arteritis, one patient had aortic insufficiency. Clinical features of temporal arteritis were absent in two patients. Temporal artery biopsy, performed in two cases, was positive. Two patients were treated with corticosteroids, associated in one case with immunosuppressive agent. Surgery was necessary in all patients. CONCLUSION: Thoracic aorta involvement in Horton's disease is most often discovered when corticotherapy is decreased. Aneurysm rupture or aortic dissection in aortic affection are the major complication. Patient with thoracic aorta involvement need a clinical and radiological follow up over long period. PMID- 14744646 TI - [Ultra-late metastasis of ocular melanoma: about two cases]. AB - INTRODUCTION: Ocular melanoma is associated with extended risk of metastases, mainly liver metastases. Nevertheless, the late and ultra-late recurrence (respectively 10 and 15 years after initial treatment) are exceptional events. EXEGESIS: We describe hereafter two cases. In the first case, the treatment is palliative chemotherapy with death 12 months after metastasis diagnosis. In the second case, the patient underwent two resections of metastases (lung and liver). This patient had excellent quality of live, and survived 11 months later after palliative surgery. DISCUSSION: Follow-up of ocular melanoma patients should include liver enzymes and ultrasonography of liver in the long run. PMID- 14744647 TI - [Delayed diagnosis of homocystinuria by major deficiency in cystathionine beta synthase]. AB - INTRODUCTION: Homocystinuria due to cystathionine beta synthase (CBS) deficiency is a special type of hyperhomocysteinemia because of its clinical expression (thrombotic events, ectopic lens and mental retardation). It's a rare, hereditary recessive autosomic disease generally diagnosed during childhood. EXEGESIS: Thrombophilia examination in a 50-year-old man found a dramatically increase homocysteinemia. Homocystinuria, profile of plasmatic amino acids and reduced CBS activity, (0.05 microkat/kg protein; N = 1.5 +/- 0.8) confirmed homocystinuria's diagnosis. Family study demonstrates that three siblings suffer from homocystinuria. Vitamin enriched diet with pyridoxin, vitamin B12 and folates induced reducing hyperhomocysteinemia and homocystinuria. CONCLUSION: This case report, original because of the diagnosis age, suggests a hyperhomocysteinemia's screening in patients with recurrent thrombotic events. PMID- 14744648 TI - [Sneddon-Wilkinson disease. Four cases report]. AB - INTRODUCTION: We report four cases of subcorneal pustular dermatosis or Sneddon Wilkinson disease. Clinical and histological lesions and immunofluorescence data were presented. This disease is classified among neutrophilic dermatitis. PATIENTS AND METHODS: All of four patients presented with clinical and histological lesions compatible with the diagnosis of Sneddon-Wilkinson disease. Indeed, direct and indirect immuno-testing were negative. We noted an association with a benign IgA monoclonal gammapathy in one case and with a seronegative polyarthritis in one other case. Three patients correctly responded to dapsone. One of them after transient improvement was resistant to dapsone and then dramatically responded to etretinate. CONCLUSION: Subcorneal pustular dermatosis is a chronic disease, rarely described in literature. It's a pustular eruption, involving the trunck, axillae and inguinal holds. It's often associated with monoclonal gammapathy, particulary IgA. Its nosological situation is still contested, especially with IgA pemphigus sharing with it the association with IgA monoclonal gammapathy and the same efficacy of dapsone. We discuss relationships between both diseases. PMID- 14744649 TI - [Emphysematous cystitis]. PMID- 14744650 TI - [Sudden death due to primary systemic amyloidosis]. PMID- 14744651 TI - [Severe typhoid fever and hemophagocytic syndrome]. PMID- 14744652 TI - [Auricular chondritis and Wegener's granulomatosis: a rare association but doubtless not fortuitous]. PMID- 14744653 TI - [Legionellosis in patient treated with infliximab]. PMID- 14744654 TI - [Subacute dysphagia in elderly woman: an unexpected foreign body]. PMID- 14744655 TI - [American College of Rheumatology, October 23-28, 2003, Orlando, Florida]. PMID- 14744656 TI - Patterns of contraceptive use within teenagers' first sexual relationships. AB - CONTEXT: Teenagers have a high unintended pregnancy rate, in part because of inconsistent use or nonuse of contraceptives. It is important to determine how partner and relationship characteristics are related to contraceptive use and consistency within adolescents' first sexual relationships. METHODS: Logistic and multinomial logistic regression analyses of data from 1,027 participants in the first two waves of the National Longitudinal Study of Adolescent Health examined the influence of relationship and partner characteristics on ever-use and consistent use of contraceptive methods during teenagers' first sexual relationships. RESULTS: Teenagers who had waited a longer time between the start of a relationship and first sex with that partner, discussed contraception before first having sex or used dual contraceptive methods had significantly increased odds of ever or always using contraceptives. Adolescents who had taken a virginity pledge, had an older partner, had a greater number of close friends who knew their first partner, or reported having a relationship that was not romantic but that involved holding hands, kissing and telling their partners they liked or loved them had decreased odds of contraceptive use or consistency. As relationship length increased, teenagers were more likely to ever have used a method, but less likely to always have used a method. CONCLUSIONS: Parents and programs should encourage teenagers to delay sexual intercourse, discuss contraception with partners before initiating sex and be vigilant about contraceptive use, particularly in long-term sexual relationships and in relationships with older partners. PMID- 14744657 TI - Partner influence on early discontinuation of the pill in a predominantly Hispanic population. AB - CONTEXT: Although studies have examined U.S. pill users' patterns of discontinuation, little is known about pill discontinuation, and the effects of partner influence, among Hispanics. METHODS: Follow-up data on pill use were collected from 213 predominantly Hispanic women who requested the pill in an urban family planning clinic in 2000. Logistic regression analyses were conducted to assess the association between a range of factors, including partner and relationship variables, and the odds of discontinuation within one month. RESULTS: The overall discontinuation rate within one month was 23%. Women who reported that their partner was unaware of their planned pill use had significantly elevated odds of discontinuing use (odds ratio, 3.4). Other variables that were independently associated with increased odds of early pill discontinuation were not taking the first pill during the clinic visit (3.0), feeling happy about the prospect of a pregnancy in the next six months (2.4) and intending to use the pill for one year or less (2.3). Age, which was assessed as a continuous variable, was negatively associated with the odds of early discontinuation (0.9). CONCLUSIONS: Male partners' awareness of planned pill use may be a marker for the level of communication and commitment in the relationship. Women may choose to disclose their planned pill use to supportive partners, and this may be beneficial to the relationship and the couple's contraceptive use. However, clinicians should take women's circumstances and needs into account before counseling them to tell a partner that they intend to use the pill. PMID- 14744658 TI - Factors associated with the content of sex education in U.S. public secondary schools. AB - CONTEXT: While sex education is almost universal in U.S. schools, its content varies considerably. Topics such as abstinence, and basic information on HIV and other sexually transmitted diseases (STDs), are commonly taught; birth control and how to access STD and contraceptive services are taught less often. Factors potentially associated with these variations need to be examined. METHODS: Data on 1,657 respondents to a 1999 national survey of teachers providing sex education in grades 7-12 were assessed for variation in topics covered. Logistic regression was used to ascertain factors associated with instruction on selected topics. RESULTS: The content of sex education varied by region and by instructors' approach to teaching about abstinence and contraception. For example, teaching abstinence as the only means of pregnancy and STD prevention was more common in the South than in the Northeast (30% vs. 17%). Emphasizing the ineffectiveness of contraceptives was less common in the Northeast (17%) than in other regions (27-32%). Instructors teaching that methods are ineffective and presenting abstinence as teenagers' only option had significantly reduced odds of teaching various skills and topics (odds ratios, 0.1-0.5). CONCLUSIONS: Instructors' approach to teaching about methods is a very powerful indicator of the content of sex education. Given the well-documented relationship between what teenagers learn about safer sexual behavior and their use of methods when they initiate sexual activity, sex education in all U.S. high schools should include accurate information about condoms and other contraceptives. PMID- 14744659 TI - A new look at an old method: the diaphragm. PMID- 14744660 TI - Comparative subchronic inhalation study of smoke from the 1R4F and 2R4F reference cigarettes. AB - A subchronic, nose-only inhalation study compared the effects of mainstream smoke from a 1R4F research cigarette to that of a 2R4F research cigarette. Male and female rats were exposed for 1 h/day, 5 days/wk, for 13 wk to mainstream smoke at 0, 0.06, 0.20, or 0.80 mg wet total particulate matter per liter of air. Clinical signs, body and organ weights, clinical chemistry, hematology, carboxyhemoglobin, serum nicotine, pulmonary plethysmography, gross pathology, and histopathology were determined. When histological changes resulting from exposure to smoke from the two types of cigarettes were compared, no biologically significant differences were observed. At the end of the exposure period, subsets of rats from each group were maintained without smoke exposures for an additional 13 wk (recovery period). At the end of the recovery period, there were no statistically significant differences in histopathological findings observed between the 1R4F and the 2R4F cigarettes. The complete toxicological assessment in this comparative inhalation study of 1R4F and 2R4F cigarettes suggests no overall biologically significant differences between the rats exposed to the two cigarettes. PMID- 14744661 TI - Influence of relatively low level of particulate ar pollution on hospitalization for COPD in elderly people. AB - To assess the association between relatively low levels of size-fractioned particulate matter (PM) and hospitalization for chronic obstructive pulmonary disease (COPD), we conducted a time-series analysis among elderly people 65 yr of age or more living in Vancouver between June 1995 and March 1999. Measures of thoracic PM (PM(10)), fine PM (PM(2.5)), coarse PM (PM(10-2.5)), and coefficient of haze (COH) were examined over periods varying from 1 to 7 days prior to hospital admissions. Generalized additive models (GAMs; general linear models, GLMs) were used, and temporal trends and seasonal and subseasonal cycles in COPD hospitalizations were removed by using GLM with parametric natural cubic splines. The relative risks were calculated based on an incremental exposure corresponding to the interquartile range of these measures, and were adjusted for daily weather conditions and gaseous pollutants. PM measures had a positive effect on COPD hospitalization, especially 0 to 2 days prior to the admissions, before copollutants were accounted for. For 3-day average levels of exposure the relative risk estimates were 1.13 (95% confidence interval: 1.05-1.21) for PM(10), 1.08 (1.02-1.15) for PM(2.5), 1.09 (1.03-1.16) for PM(10-2.5), and 1.05 (1.01-1.09) for COH. The associations were no longer significant when NO(2) was included in the models. We concluded that the particle-related measures were significantly associated with COPD hospitalization in the Vancouver area, where the level of air pollution is relatively low, but the effects were not independent of other air pollutants. PMID- 14744662 TI - Lung tumor response in strain a mice exposed to tobacco smoke: some dose-effect relationships. AB - Male strain A/J mice were exposed for 5 mo in a whole-body inhalation chamber to 3 different concentrations of a mixture of cigarette sidestream and mainstream smoke (99, 120, and 176 mg/m(3) of total suspended particulate material, TSP). After an additional 4-mo recovery period in air, lung tumor multiplicities and incidences were determined. The two highest smoke concentrations produced significantly more lung tumors than did the low dose and control groups, although the response to the high was slightly less than to the medium dose. Lung tumor incidences were in all three groups significantly higher than in controls. Lung displacement volume was increased in a dose-dependent manner, but morphometric analysis of the tissues failed to provide evidence for airspace enlargement. Plasma cotinine levels were dose-dependent and similar after 1-day and 5-day exposure. The shape of the dose-response curve and a comparison with previous data suggest that cigarette smoke is only a comparatively weak mouse lung carcinogen. PMID- 14744663 TI - Ovalbumin-induced airway inflammation and fibrosis in mice also exposed to ozone. AB - A murine model of allergen-induced airway inflammation was used to examine the effects of exposure to ozone on airway inflammation and remodeling. Sensitized BALB/c mice were exposed to ovalbumin aerosol for 4 wk before and after 2 wk of exposure to either 0.2 ppm or 0.5 ppm ozone. Other groups of mice were exposed to ovalbumin aerosol for 6 wk with continuous concurrent exposure to ozone during wk 1-6, or during intermittent concurrent exposure to ozone. Lung inflammation was measured by quantitative differential evaluation of lung lavage cells and by histological evaluation of stained lung sections. Alterations in lung structure (airway fibrosis) were evaluated by quantitative biochemical analysis of microdissected airways. The same total number of cells was observed in lavage fluid from animals exposed for 4 wk to ovalbumin alone or to ovalbumin for 4 wk immediately before or after exposure to 2 wk of 0.2 or 0.5 ppm ozone. Mice exposed to ovalbumin for 6 wk with concurrent exposure to either 0.2 ppm or 0.5 ppm ozone during wk 3-6 had a significant decrease in the total number of cells recovered by lavage. Values as low as 7% of the cell number found in mice exposed to ovalbumin aerosol alone were observed in the mice exposed to ovalbumin plus 0.2 ppm ozone during wk 3-6. There were significant differences in the cell differential counts in the lavage fluid from mice exposed to ovalbumin alone as compared with values from mice exposed to ovalbumin and ozone under all of the protocols studied. When ozone was given for 2 wk prior to ovalbumin exposure (Experiment 1), there were a high percentage of macrophages and low percentages of lymphocytes and eosinophils in the lung lavage. When ozone was given for 2 wk after ovalbumin exposure (Experiment 2), there were a moderate percentage of macrophages, a low percentage of eosinophils, and a high percentage of lymphocytes in the lung lavage. When ozone and ovalbumin were given simultaneously (Experiments 3 and 4), there were a high percentage of macrophages in the lavage with 0.2 ppm ozone and a high percentage of eosinophils. Ozone appears to antagonize the specific inflammatory effects of ovalbumin exposure, especially when given before or during exposure to ovalbumin. Airway remodeling was examined by two different quantitative methods. None of the groups exposed concurrently to ovalbumin and ozone had a significant increase in airway collagen content as compared to the matched groups of mice exposed to ovalbumin alone. The findings were consistent with an additive response of mice to simultaneous exposure to ovalbumin and ozone. Ozone exposure alone for 6 wk did not affect the number of goblet cells in the airways, while mice exposed to ovalbumin aerosol alone for 6 wk had about 25% goblet cells in their conducting airways. Concurrent exposure to ovalbumin and 0.2 ppm ozone caused significant increases in goblet cells (to 43% of total cells) in the conducting airways of the exposed mice. We conclude that when mice with allergen-induced airway inflammation induced by ovalbumin are also exposed to ozone, the lung inflammatory response may be modified, but that this altered response is dependent on the sequence of exposure and the concentration of ozone to which they are exposed. At the concentrations of ozone tested, we did not see changes in airway fibrosis. However, goblet-cell hyperplasia appeared to be increased in mice exposed concurrently to ovalbumin and 0.2 ppm ozone. PMID- 14744664 TI - A novel method to aerosolize powder for short inhalation exposures at high concentrations: isolated rat lungs exposed to respirable diesel soot. AB - More efficient methods are needed to aerosolize dry powders for short-duration inhalation exposures at high concentrations. There is an increasing need to reach the peripheral lung with dry powder medications as well as with collected ambient aerosol particulates in environmental research projects. In a novel aerosol generator, a fixed volume of compressed air was used to create a short burst of a highly concentrated aerosol in a 300-ml holding chamber. Collected diesel soot was deagglomerated to a fine aerosol with a mass median aerodynamic diameter (MMAD) of 0.55 microm, not much larger than the 0.25 microm MMAD of diesel exhaust particles measured in air. A fine powder such as 3-microm silica particles was completely deagglomerated to an aerosol with a MMAD of 3.5 microm. Immediately after generation, the aerosol was available for exposure at a chosen flow rate by the use of an automated valve system. Tritium-labeled diesel soot was thus used to expose the isolated perfused rat lung at an air concentration of approximately 3 mg/L and a flow rate of 370 ml/min in a 1-min-long exposure. The lungs were ventilated at 75 breaths/min and a tidal volume of 1.13 +/- 0.11 ml (SD, n = 3). Results showed that 19.8 +/- 1.1 microg (SD, n = 3) soot was deposited in the lungs. This amount constitutes 9.5% of the amount inhaled and is close to literature data on deposition of similar sized particles in the rat lung. More than 97% of the deposited soot was located distal to the extrapulmonary bronchi, indicating that the system delivers a highly respirable aerosol. The aerosol system is particularly useful for peripheral lung delivery of collected ambient aerosols or dry powder pharmaceuticals following a minimal effort in formulation of the powder. PMID- 14744665 TI - Exposure to concentrated ambient particles (CAPs): a review. AB - Epidemiologic studies support a participation of fine particulate matter (PM) with a diameter of 0.1 to 2.5 microm in the effects of air pollution particles on human health. The ambient fine particle concentrator is a recently developed technology that can enrich the mass of ambient fine particles in real time with little modification. The advantages of concentrators are that the particles produced are "real world" and they allow exposure at pertinent masses. Limitations include variability in both particle mass and composition and some uncertainty over the best statistical approach to analyze the data. Cumulative evidence provided by the body of initial investigation shows that exposures to concentrated ambient particles (CAPs) can be accomplished safely in both humans and animals. Human investigation using the CAPs has shown acute lung inflammation and changes in both blood indices and heart rate variability. Animal studies support a potential pulmonary inflammation, blood changes, alterations of specific cardiac endpoints, and an increased susceptibility of specific models. These studies have helped establish the causal relationship between find particle exposure and adverse health effects in the lung and cardiovascular system. In addition, it appears that specific components in CAPS may differentially affect these tissues. PMID- 14744666 TI - Diesel exhaust-associated gas components enhance chemokine production by cervical lymph-node cells from mice immunized with sugi basic proteins. AB - Previously, we showed that exposure to diesel exhaust (DE) increased inflammatory cells in the airway and cytokine production from local lymph-node cells after antigen stimulation. To clarify the role of particle-free diesel gas components in induction of allergic inflammation, we compared the effect of DE and gas components on pollen-antigen-stimulated chemokine production by cervical lymph nodes (CLN) cells in BALB/c mice. Groups of mice were exposed to 0 (control), 1.0 mg diesel exhaust particles (DEP)/m(3) (DE), or filtered 1.0 mg DEP/m(3) DE (gas) for 12 h daily for 5 wk. Each group of mice was injected intraperitoneally with sugi basic protein (SBP), a major allergen of Japanese cedar pollen, immediately before their exposure to DE or gas. On days 14 and 35, each mouse received an additional SBP intranasally. Exposure to DE or gas did not affect the lymphocyte subpopulations of CLN. Culture supernatants of CLN cells from DE-exposed, SBP immunized mice had significantly increased levels of monocyte chemoattractant protein-1. Exposure to gas significantly increased the amount of thymus- and activation-regulated chemokine and macrophage inflammatory proteins-1 alpha in the CLN cells from SBP-immunized mice. These results suggest that Gas components as well as DEP may differentially regulate production of chemokines at local sites. PMID- 14744668 TI - Neurocognitive sequelae of childhood cancers and their treatment. PMID- 14744670 TI - Feuerstein, R., Rand, Y., and Hoffman, M. et al.: Cognitive modifiability in retarded adolescents: effects of Instrumental Enrichment. American Journal of Mental Deficiency 83:539-550, 1979. PMID- 14744671 TI - Cognitive modifiability in retarded adolescents: effects of Instrumental Enrichment. 1979. PMID- 14744672 TI - Research in paediatric neuropsychology--past, present and future. AB - Reviews of the research literature in a range of neurodevelopmental disorders and acquired brain injury reveal a remarkably consistent historical transition through three phases, here termed structural, theoretical and dynamic neuropsychology. Of course, any attempt to summarize such a complex and rich history using a simplistic heuristic will inevitably fail to capture the wide diversity of the research effort. Nevertheless, it is argued that looking at three distinct phases in the history of research helps to organize the field and points to possible future directions for applied research. Using examples from an eclectic range of disorders including childhood obsessive compulsive disorder, congenital hemiplegia and disorders implicating mutation of neurodevelopmental control genes, the implications for future efforts in paediatric neurorehabilitation are considered. PMID- 14744673 TI - Improved ambulation and speech production in an adolescent post-traumatic brain injury through a therapeutic intervention to increase postural control. AB - PURPOSE: This case study examined the effectiveness of a programme designed to improve anticipatory postural control in an adolescent over years 2 and 3 post traumatic brain injury (TBI). It was hypothesized that her difficulty in walking and talking simultaneously was caused by excessive co-activation of extremity, trunk, and oral musculature during upright activities. METHODS: The participant was treated weekly by physical and speech therapy. Treatment focussed on improving anticipatory postural control during gross motor activities in conjunction with oral-motor function. RESULTS: Initially, the participant walked using a walker at a speed of 23 cm s(-1). Two years later, she could walk without a device at 53 cm s(-1). Initial laryngoscopic examination showed minimal movement of the velum or pharyngeal walls; full movement was present after treatment. The measure of intelligibility improved from no single word intelligible utterances to 85% intelligible utterances after 2 years. DISCUSSION: The results suggest that less compensatory rigidification of oral musculature was needed to maintain an upright position against gravity as postural control improved. CONCLUSION: An adolescent 1-year post-TBI was followed as she underwent additional rehabilitation focussed on improving anticipatory postural control. The functional goal of simultaneously talking while walking was achieved through this intervention. PMID- 14744674 TI - Abstracts of the International Conference on Conservative Management of Spinal Deformities. Barcelona, Spain, 23-24 January 2004. PMID- 14744677 TI - Methods for allergen analysis in food: a review. AB - Food allergies represent an important health problem in industrialized countries. Undeclared allergens as contaminants in food products pose a major risk for sensitized persons. A proposal to amend the European Food Labelling Directive requires that all ingredients intentionally added to food products will have to be included on the label. Reliable detection and quantification methods for food allergens are necessary to ensure compliance with food labelling and to improve consumer protection. Methods available so far are based on protein or DNA detection. This review presents an up-to-date picture of the characteristics of the major food allergens and collects published methods for the determination of food allergens or the presence of potentially allergenic constituents in food products. A summary of the current availability of commercial allergen detection kits is given. One part of the paper describes various methods that have been generally employed in the detection of allergens in food; their advantages and drawbacks are discussed in brief. The main part of this review, however, focuses on specific food allergens and appropriate methods for their detection in food products. Special emphasis is given to allergenic foods explicitly mentioned in the Amendment to the European Food Labelling Directive that pose a potential risk for allergic individuals, namely celery, cereals containing gluten (including wheat, rye and barley) crustaceans, eggs, fish, peanuts, soybeans, milk and dairy products, mustard, tree-nuts, sesame seeds, and sulphite at concentrations of at least 10 mg kg(-1). Sulphites, however, are not discussed. PMID- 14744678 TI - Flow-through spectrophotometric sensor for the determination of saccharin in low calorie products. AB - A simple, rapid and inexpensive monoparameter flow-through sensor has been developed for the determination of saccharin in low calorie and dietary products. The method is based on the transient adsorption of the sweetener on Sephadex G-25 solid phase packed to a height of 20 mm in the flow cell. The optimal transient retention of the synthetic sweetener, in terms of sensitivity and sampling frequency, was obtained when pH 2.75 citric acid-sodium citrate buffer 5 x 10(-3) M was used as a carrier at a flow-rate of 1.5 ml min(-1). Saccharin was determined measuring its intrinsic absorbance at 217 nm at its residence time. Calibration graphs for peak height and peak area were linear over the range 5.0 200.0 microg ml(-1), RSD 1.18%, and 1.0-200.0 microg ml(-1), RSD 0.78%, respectively. Saccharin was determined in several food samples measuring height or area peak, obtaining recoveries ranging between 98-104 and 99-102% for height and area peak, respectively. The procedure was validated for use in the determination of saccharin in low calorie and dietary products giving reproducible and accurate results. PMID- 14744679 TI - Phytosterols: lack of cytotoxicity but interference with beta-carotene uptake in Caco-2 cells in culture. AB - Ingestion of phytosterols has been shown to reduce plasma cholesterol in both animals and humans. The esterified forms of phytosterols are increasingly being incorporated into margarine and fat spreads, which are then marketed as functional foods. The aim was to assess the cytotoxicity and uptake of four phytosterols, beta-sitosterol, campesterol, stigmasterol and stigmastanol, in human intestinal cells in culture. Another aim was to determine if phytosterols would interfere with alpha-tocopherol or beta-carotene uptake by these cells. Human adenocarcinoma Caco-2 cells were supplemented for 24 h with increasing concentrations (0-12.5 microM) of each phytosterol. Cytotoxicity was assessed by neutral red uptake (NRU), lactate dehydrogenase release (LDH) and fluorescein diacetate/ethidium bromide (FDA/EtBr) assays. The phytosterols had no significant effects on Caco-2 cell viability assessed using LDH and FDA/EtBr assays. The highest concentrations of beta-sitosterol and campesterol tested (12.5 microM) resulted in decreased cell viability assessed using the NRU assay. All phytosterols were taken up by Caco-2 cells in culture. The results demonstrate a reduction in the uptake of beta-carotene when Caco-2 cells were supplemented with 20 microM beta-sitosterol. beta-Sitosterol did not interfere with alpha tocopherol uptake by the cells. In conclusion, Caco-2 cells are a useful model system to study potential interactive effects of phytosterols with fat-soluble dietary components. PMID- 14744680 TI - Confirmatory assay for zeranol, taleranol and the Fusarium spp. toxins in bovine urine using liquid chromatography-tandem mass spectrometry. AB - A method is described for the quantitative determination of the veterinary drug zeranol, its epimer taleranol and the mycotoxins zearalenone, alpha-zearalenol and beta-zearalenol in bovine urine. The method is based on liquid chromatography coupled to negative-ion electrospray mass spectrometry-mass spectrometry of urine extracts prepared by solid-phase extraction with C(18) columns. Two transition ions at m/z 277 and 91 are monitored for zeranol and taleranol along with the transition ion at m/z 281 for their respective deuterated (d(4)) internal standards. Similarly, two transitions are monitored for each of the three mycotoxins along with a transition ion for each of their corresponding internal standards. The method has been validated according to the new European Union criteria for analysis of veterinary drug residues, and is suitable for monitoring urine samples taken under National Surveillance Schemes. The method has been validated at 1, 1.5 and 2 ng ml(-1) for zeranol and taleranol and at 5, 10 and 15 ng ml(-1) for each of the three mycotoxins. Correlation between the described method and a routine method, based on gas chromatography-mass spectrometry, was assessed using a range of naturally incurred samples. PMID- 14744681 TI - Implications of the use of semicarbazide as a metabolic target of nitrofurazone contamination in coated products. AB - Data from the Brazilian Agricultural Ministry show that before the implementation of the Brazilian programme of nitrofuran control in February 2003, the cases of contamination of Brazilian chicken by nitrofurans were almost exclusively due to furaltadone. After May 2003, such cases decreased until no more reports of Brazilian chicken contamination with this nitrofuran were reported. Curiously, after April 2003, an increase was observed in the numbers of contaminated samples by semicarbazide, the target metabolite of nitrofurazone. Most Brazilian chicken found to be contaminated with semicarbazide has been covered with flour, salt and spices. For this reason, the authors' laboratory initialized a programme for tracing possible sources of food contamination by semicarbazide. After several semicarbazide positives in flour of controlled origin (results varying between 2.2 and 5.2 microg kg(-1)), the different additives used in the cereal industry as flour improvement agents were studied. The results indicate that the compound azodicarbonamide was responsible for the source of the contaminant semicarbazide. PMID- 14744682 TI - Estimation of the daily exposure of Koreans to aflatoxin B1 through food consumption. AB - A survey for aflatoxin B(1) (AFB(1)) was conducted on 88 food-grade rice samples randomly collected during July and August 2002 in Seoul, Korea. The presence of AFB(1) was determined by enzyme-linked immunosorbent assay, and the positive samples from enzyme-linked immunosorbent assay were confirmed using high performance liquid chromatography. Besides this, from the surveying data from the literature published since 1997, the intake of AFB(1) from food in Korea was calculated and compared with the provisional maximum tolerable daily intakes. Naturally occurring AFB(1) was found in 5/88 (6%) samples of rice with an average of 4.8 ng g(-1). A calculated probable daily intake of AFB(1) for Koreans fell into the range 1.19-5.79 ng kg(-1) bw day(-1), hence exceeding the estimated provisional maximum tolerable daily intakes. In conclusion, the exposure of Koreans to AFB(1) could bring about health concerns. This is the first report discovering that rice is the major contributor to the dietary intake of AFB(1) in Korea. PMID- 14744683 TI - Atmospheric pressure photo-ionization liquid chromatography/mass spectrometric determination of aflatoxins in food. AB - A comparison between electrospray ionization (ESI) and the recent introduced atmospheric pressure photo-ionization (APPI) technique is described for the liquid chromatography/mass spectrometric (LC/MS) determination of aflatoxin B1, B2, G1 and G2 in food. For the optimization of APPI, several APPI ion source parameters were examined. Furthermore, the chemical noise and signal suppression of ESI and APPI by sample matrix interference were investigated. The results indicated that APPI has the low chemical noise and the little signal suppression in comparison with ESI. The mean recovery of each aflatoxin from peanuts, corn, nutmeg and red pepper sample spiked at 1 ng g(-1) ranged from 89 to 105% with relative standard deviations of 3.3-5.1%. The limit of detection (S/N = 3) in all food was in the range 0.11-0.5 ng g(-1). From these studies, it is suggested that APPI can be used as an additional tool in food residue analysis. PMID- 14744684 TI - Determination of ochratoxin A in virgin olive oils of Greek origin by immunoaffinity column clean-up and high-performance liquid chromatography. AB - An improved specific analytical method for ochratoxin A (OTA) determination in olive oil is described, using a methanolic-aqueous extraction, an immunoaffinity column clean up step and high-pressure liquid chromatography with fluorescence detection. The mean recovery was found at 108% (relative standard deviation, RSD = 4.7%) and the detection limit (DL) was estimated at 4.6 ng kg(-1). Along with OTA, aflatoxin B(1) (AFB(1)) was determined using the same extract. The recovery factor was 84.8% (RSD = 17.8%) and the DL was 56 ng kg(-1) olive oil. Both determinations were applied in 50 samples of olive oil originated from representative regions of Greece. Results revealed the presence of OTA in 88% of samples tested (n = 44, mean 267 ng kg(-1)). Among them, 10 were contaminated with more than 500 ng kg(-1) (median 568 ng kg(-1)), 10 with 200-500 ng kg(-1) (median 260 ng kg(-1)), 15 with 100-200 ng kg(-1) (median 140 ng kg(-1)), nine with DL-100 (median 60 ng kg(-1)) and in six samples, OTA was not detectable. Interestingly, most contaminated samples were from Southern Greece. Results of AFB(1) determination showed the presence of aflatoxin B(1) (60 ng kg(-1)) in only one olive oil sample also from Southern Greece. The levels of OTA found in Greek olive oil were relatively low as compared with other commodities such as cereals or wine reported in the literature. PMID- 14744686 TI - Medical education and health services research: the linkage. AB - The medical community is coming under increased scrutiny. Challenges to the integrity of the healthcare system have been raised due to reports about the prevalence of medical errors. A heightened level of vigilance is required. Equally important is the need to isolate and correct the source of any problem, perceived or real. We are faced with challenging questions. Is the selection of students and residents appropriate? Are their education and evaluation valid? These questions must be answered at least in part by understanding the climate in which the services to the patients are rendered. Otherwise deficiencies noted in practice may be inappropriately attributed to the educational process. This article addresses the importance, implications and impact of the link between medical education and health services research. The goal of medical education is to prepare physicians to meet the challenges of practice by fulfilling their roles of clinician, educator and resource manager. Health services research can be linked to any of these physician roles. An understanding of health services is necessary to assess how well this goal is being met in the context of the changing healthcare system. A partnership between medical education and health services research is essential for academic health centers and health services institutions in assessing issues of health manpower and for the public good. Academic health centers have an important role in this partnership providing an infrastructure and expertise for both education and health services research. PMID- 14744687 TI - Becoming a student in PBL course: twelve tips for successful group discussion. AB - Problem-based learning (PBL) serves as an educational method to foster self directed learning, integration across disciplines, small-group learning and decision-making strategies. The approach is student centred. During the discussion of a PBL case there are a number of important issues to be considered by students, such as keeping ground rules, knowing their roles, keeping group dynamics, becoming a purposeful learner, planning how to use tutors' feedback to enhance group discussion and boost student's learning skills, as well as striving to become a winning team. This paper provides 12 practical tips to PBL students to enhance their skills in discussing a case in their group. PMID- 14744688 TI - Developing the teaching instinct. 11: Small group learning. PMID- 14744689 TI - How to...use audiovisual aids--a 25-year update. PMID- 14744690 TI - Clinical teachers' tacit knowledge of basic pedagogic principles. AB - Academic faculty members in medical schools rarely receive formal instruction in basic pedagogic principles; nevertheless many develop into competent teachers. Perhaps they acquire tacit knowledge of these principles with teaching experience. This study was designed to assess clinical teachers' tacit knowledge of basic pedagogic principles and concepts. The authors developed a multiple choice question (MCQ) exam based on 20 pedagogic principles judged by a panel of education experts to be important for clinical teaching. Three groups of clinician-educators sat the test: (1) clinicians with advanced education training and experience; (2) internal medicine specialists; (3) surgical specialists. All four groups of clinicians-educators passed the test, indicating that they possess a reasonable tacit knowledge of basic pedagogic principles. Those with advanced education training performed much better than members of the other two groups while specialists and residents working in teaching hospitals outperformed specialists from non-teaching hospitals. It is possible that converting this tacit knowledge to explicit knowledge may improve individual teaching effectiveness. PMID- 14744691 TI - Formal appraisal of undergraduate medical students: is it worth the effort? AB - Medical student stress is most often related to difficulties of adjusting to university academic standards, and work-social life balance. Faculty systems identify academically failing students for counselling, whilst the majority of students do not have opportunities for individual discussion about progress. This study reports a pilot formal appraisal process for first-year undergraduates. Preparatory material required students to reflect on their academic performance, factors contributing to their university life and satisfaction with career choice. Individual appraisal sessions were held with trained, experienced senior faculty staff, with completion of an appraisal record to document agreed outcomes. Individualized study skills advice was the commonest documented outcome on appraisal records. Students were overwhelmingly positive about the experience, reporting both enhanced perceptions of faculty and reduced anxiety about academic performance. Medical schools have responsibilities to consider ways to optimize students' performance; attainment can be related more to personal and motivational factors than academic ability. PMID- 14744692 TI - Student-selected components in the undergraduate medical curriculum: a multi institutional consensus on purpose. AB - Student-selected components (SSCs) are an established part of undergraduate medical curricula in the UK. Implementation has been against a background of differing educational approaches to curricular change, together with a lack of clarity about their purpose, relationship with the core curriculum and contribution to overall assessment. This has resulted in a diversity of programmes with perceived differing importance between medical schools. This paper documents the approach used by a consortium of medical schools with diverse curricula to develop consensus on the educational purpose and learning outcomes of SSCs. Agreement on common purposes and outcomes was achieved, and consensus documents are reported. These may be valuable for other medical schools implementing any form of student-selected project work. This work will now be the starting point for further work on producing recommendations for assessment of SSCs, which will be applicable across different medical schools. PMID- 14744693 TI - Identifying the perceived weaknesses of a new curriculum by means of the Dundee Ready Education Environment Measure (DREEM) Inventory. AB - The Canadian Memorial Chiropractic College is in the fourth year of implementing a new four-year Integrative Curriculum. In order to determine how the students experienced the educational environment generated by the new curriculum, the Dundee Ready Education Environment (DREEM) Inventory was administered on the same day to 148 (96%) first-year, 131 (87%) second-year, and 128 (84%) third-year students (n = 407). Resultant scores indicated many areas of concern. However, bimodal and large numbers of 'uncertain' responses also occurred. In order to establish and compare areas of most concern between the three year groups, responses were calculated as percentages indicating for each item those who agree, disagree or are uncertain. This result clearly indicated areas of joint concern for each of the five domains addressed by the DREEM Inventory. Analysis of these concerns formed the basis of strategic planning in order to institute remedial action, and on which to focus institutional resources. PMID- 14744694 TI - A self-directed summative examination in problem-based learning in dentistry: a new approach. AB - This paper describes and evaluates a new method of assessment in PBL, which was developed with two cohorts of dental students. The method involves students in PBL groups designing PBL problems and assessment tasks that are in line with the objectives of the course and the principles of PBL. Construction of tasks and feedback were provided. The student groups based the summative assessment on a selection of the PBL assessments provided. The evaluation indicates that senior students are capable of designing challenging PBL assessment tasks that are in line with course objectives and PBL principles. This experience raised the pass rates of the students compared those of with earlier cohorts of students. The students rated the method favourably; they considered it enabled them to demonstrate their competences and that it was more closely in line with PBL principles than other methods of assessment that they had experienced. The method is a promising innovation, which could be used in a variety of PBL courses. PMID- 14744695 TI - Student attitudes and their academic performance: is there any relationship? AB - This study examined the relationship between students' attitudes to consultation skills, their confidence in performing those skills and their academic results. Final-year medical students completed a questionnaire before and after the year 6 general practice attachment, which teaches holistic, patient-centred clinical management and emphasizes core verbal and non-verbal communication skills. When underlying academic ability was taken into account only pre-attachment attitudes regarding the importance of consultation skills remained significantly correlated with at least one measure of performance. Student confidence in performing a skill was not associated with academic measures of performance. These results show that students perform better on those skills that they value and this may be influenced by underlying motivation to master the skill. In contrast, students' confidence in performing a skill is not related to their assessed performance and using confidence as a performance measure may misrepresent the quality of learning being assessed. PMID- 14744696 TI - Demonstration of the effectiveness and acceptability of self-study module use in residency education. AB - Educators face increasing challenges to promote lifelong learning skills, to include new content areas in an already full curriculum and to maximize limited resources for curriculum implementation. Self-study modules (hereafter modules) offer potential solutions. Three modules on preventive medicine topics were evaluated in Family Medicine residencies. A retrospective pre-/post-test of a resident's ability to meet the module's objectives was used for evaluation. Additionally, residents rated the appropriateness and acceptability of the modules, their preference for 13 methods of learning, and completed a multiple choice knowledge test. This study demonstrates the effectiveness of modules at multiple levels of evaluation in accordance with a modified version of Kirkpatrick's hierarchy of levels of evaluation. Residents found the modules to be acceptable and useful. Significant gains were seen in residents' abilities to meet objectives. The multiple-choice knowledge test was used to demonstrate mastery of the module materials at an appropriate performance level for future practitioners. Module use was in the top five choices of preferred learning methods. No correlation was seen between residents' preference for learning using modules and educational outcomes. Modules are an effective and acceptable learning method for residents. Even those who prefer other learning methods show improved educational outcomes. PMID- 14744697 TI - Linking appraisal of PRHO professional competence of junior doctors to their education. AB - In the UK, new medical graduates are known as Pre-registration House Officers (PRHOs). Postgraduate Deans are responsible for the PRHO year and for the final certification of PRHOs to allow them to be fully registered by the GMC as medical practitioners. However, as much as appraisal of professional growth is central to PRHO training, they are in need of a robust assessment mechanism to detect, at an early stage, individuals with significant clinical and professional deficiencies. Documented, reliable and valid ongoing information on PRHO performance will provide information for early intervention and will establish 'hard' observable evidence, for certification decisions. Thus, an approach that links appraisal/assessment of professionalism and clinical skills to education is the way forward. This paper describes a new approach to appraisal/assessment of PRHOs, which is currently being piloted in a number of regions in Scotland. The conceptual paradigm was developed during the last three years as a proposal for a Scottish national PRHO reform. 'Grounded' qualitative studies were employed to explore trainees' and trainers' perceptions of the expected competences (outcomes) of PRHO performance for appraisal/assessment purpose. The GMC recommendations are reviewed in light of the study results. An assessment model emerged that links appraisal to education. PRHOs' cumulative performance is documented over one year of training resulting in diagnostic profiles that provide guidance for evaluation and training of PRHOs. Poor performers are flagged in the early stages of training, thus allowing early intervention. The feasibility and acceptance of the model by educators, the health system and PRHOs has yet to be established. PMID- 14744698 TI - Clinical and education implications of shift work. AB - Yorkshire teaching hospital staffs' perceptions before and after implementing a junior doctor shift system are reported. The study is placed in context by discussing the European Working Time Directive and the New Deal for Doctors. Five education and clinical issues are examined using data from activity diaries and attitude questionnaires. Interesting findings emerge that sometimes contradict the literature. Important recommendations are made to ease the NHS's inevitable move towards shift work. PMID- 14744699 TI - A locally created EBM course for faculty development. AB - International EBM workshops have significant barriers and lack focus on institutional needs. The authors describe a local EBM curriculum and report its effectiveness and participant satisfaction. Local EBM experts devised an EBM course designed to improve faculty understanding of EBM skills and concepts. Retention of EBM concepts was assessed with a 10-item EBM knowledge test administered before and after the course. Participants were also asked to self assess five EBM skills immediately after and nine months after the course. They also filled out a satisfaction survey. In total, 61% of the participants completed the course. The EBM knowledge test showed a significant change in scores for EBM concepts. The participants' self-assessed EBM skills remained high at nine months. Participants rated most course variables highly. It is concluded that a locally developed EBM faculty curriculum can succeed if scholars define institutional needs, receive broad institutional support, use proven educational methodologies and avoid scheduling conflicts. PMID- 14744700 TI - Applying adult learning principles to medical education in the United States. AB - An extensive literature on adult learning principles has been developed over the last 40 years. Simultaneously, undergraduate medical educational programs have undergone varying degrees of curricular reform. The present paper discusses the educational and societal factors that have functioned as a catalyst for innovations in medical education, and reviews the major initiatives which have been undertaken. Data are analyzed to assess the extent to which some of these changes have been incorporated into medical curricula. A comprehensive review of adult learning principles is than presented, in order to provide a framework for the incorporation of principles of adult learning into the next wave of medical education reform. The review of adult learning principles reveals that undergraduate medical educational reform has underutilized the robust literature on adult learning. The present paper concludes with suggestions for medical education reform that incorporates the principles of adult learning and discusses the major impediment to curricular reform. PMID- 14744701 TI - Exploring medical students' attitudes towards peer physical examination. AB - With opportunities for dissection and examination of sick patients decreasing, the role of peer physical examination (PPE) is increasing. This study explores students' attitudes towards PPE and the relationship between attitudes and demographics. A total of 129 first-year medical students from the Peninsula Medical School completed the Examining Fellow Students (EFS) questionnaire. At least 97% of students were comfortable participating in PPE of all body parts except breast and inguinal regions. Over 20% of students were unwilling to participate in PPE of the breast and inguinal regions. Students were more comfortable with PPE within gender than across gender. Females were more likely to be uncomfortable with PPE. Further research with larger sample sizes is required to determine whether attitudes are related to age and religious faith. PMID- 14744706 TI - Silica deposition in relation to ageing of leaf tissues in Sasa veitchii (Carriere) Rehder (Poaceae: Bambusoideae). AB - BACKGROUND AND AIMS: Silica deposition is one of the important characteristics of plants in the family Poaceae. There have been many investigations into the distribution, deposition and physiological functions of silica in this family. Two hypotheses on silica deposition have been proposed based on these studies. First, that silica deposition occurs passively as a result of water uptake by plants, and second, that silica deposition is controlled positively by plants. To test these two apparently contradictory hypotheses, silica deposition in relation to the ageing of leaf tissues in Sasa veitchii was investigated. METHODS: Tissues were examined using a light microscope and a scanning electron microscope equipped with an energy dispersive X-ray microanalyser. KEY RESULTS: The deposition process differed depending on cell type. In mesophyll tissue, fusoid cells deposited large amounts of silica depending on leaf age after maturation, while chlorenchyma cells deposited little. In epidermal tissue, comprised of eight cell types, only silica cells deposited large amounts of silica during the leaf's developmental process and none after maturation. Bulliform cells, micro hairs and prickle hairs deposited silica densely and continuously after leaf maturation. Cork cells, guard cells, long cells and subsidiary cells deposited silica at low levels. CONCLUSIONS: The significance of these observations is discussed in relation to the two hypotheses proposed for silica deposition in Poaceae. The results of the present study clearly indicate that both hypotheses are compatible with each other dependent on cell types. PMID- 14744707 TI - Pollen dispersion, pollen viability and pistil receptivity in Leymus chinensis. AB - BACKGROUND AND AIMS: Leymus chinensis is an economically and ecologically important grass that is widely distributed across eastern areas of the Eurasian steppe. A major problem facing its propagation by man is its low sexual reproductivity. The causes of low fecundity are uncertain, largely because many aspects of the reproductive biology of this species remained unknown or incomplete. This study aims to address some of these issues. METHODS: Pollen dispersion, pollen viability, pollen longevity and pistil receptivity were studied in a representative, natural population of L. chinensis growing in Inner Mongolia. KEY RESULTS: Flowering of L. chinensis occurred at the end of June and lasted for 5 d. Pollination peaked between 1600 h and 1700 h, and about 56.1 % of the total pollen grains were released at this time. Pollen density was highest towards the middle of flowering spikes and lowest at the bottom over the 5 d measurement period. Pollen viability (62.4 %) assessed using TTC was more accurate than using IKI (85.6 %); 50 % of pollen arriving on stigmas germinated. Pollen remained viable for only 3 h and the pollen : ovule ratio was 79 333 : 1. Pistil receptivity lasted for only 3 h and, overall, 86.7 % of pistils were pollinated. Within the spike, the relative fecundity of different positions was middle > lower > upper throughout the period of pollination; daily variation of fecundity was similar to that of the pollen flow. The spikes that opened on the day of highest pollen density exhibited the highest fecundity (36.0 %). No seeds were produced by self-pollination. CONCLUSIONS: The data suggest that low pollen viability, short pollen longevity and short pistil receptivity all appear to contribute to the low seed production typical of this important forage crop. PMID- 14744708 TI - Future doctors should learn about our country's health care system. PMID- 14744709 TI - Teaching medical students and residents skills for delivering bad news: a review of strategies. AB - Although delivering bad news is something that occurs daily in most medical practices, the majority of clinicians have not received formal training in this essential and important communication task. A variety of models are currently being used in medical education to teach skills for delivering bad news. The goals of this article are (1) to describe these available models, including their advantages and disadvantages and evaluations of their effectiveness; and (2) to serve as a guide to medical educators who are initiating or refining curriculum for medical students and residents. Based on a review of the literature and the authors' own experiences, they conclude that curricular efforts to teach these skills should include multiple sessions and opportunities for demonstration, reflection, discussion, practice, and feedback. PMID- 14744710 TI - Challenging linguistic barriers to health care: students as medical interpreters. AB - Inadequate medical interpretation services are a barrier to the delivery of optimal health care to persons with limited English proficiency. Even though Title VI of the Civil Rights Act of 1964 requires that interpretation services be available to persons speaking limited English, many health care institutions are struggling to reach full compliance. Communication through untrained interpreters is likely to include mistranslations or omissions of physicians' questions, truncated or slanted patient responses, and inadequate information to facilitate accurate diagnosis and treatment. The Interpreter's Aide Program (IAP) is a service-learning program that was implemented at Brown Medical School in 1997. The IAP is a collaborative effort among Brown students, the Rhode Island Hospital Department of Social Work, and Brown Medical School. This three-way partnership strengthens the IAP and expands interpretation services to Spanish- and Portuguese-speaking patients at Rhode Island Hospital. Bilingual undergraduate and medical students become trained medical interpreters and render community service while developing cross-cultural skills. The authors review the development and implementation of the IAP. There is potential for other academic health centers to develop similar partnerships with local colleges and universities, and to provide service-learning opportunities for future physicians and health care consumers. PMID- 14744711 TI - Medical education goes to prison: why? AB - The authors describe a pilot medical education program that developed a new and ongoing correctional medicine curriculum for third- and fourth-year medical students at Nova Southeastern University College of Osteopathic Medicine. During the first two years of the pilot program (2000-02), a total of 53 students were placed in one-month rotations in prison health care settings. Students received orientations, directed readings, and prison clinic experience under the director of board-certified physician preceptors. An evaluation of the pilot experience was conducted by student survey. The findings indicate that students had positive experiences related to continuity of care, access to pathology, access to procedures, and exposure to a unique managed care model. Students requested more structured curriculum and more opportunities to develop content understanding of the unique clinical aspects of prison health care. The authors conclude that given the increasing U.S. prison population, the constitutional requirement to provide medical care to inmates, and demand for career-oriented correctional physicians, the favorable outcome of this pilot educational program provides support for implementing such programs in medical schools throughout the country. They also speculate that the program may encourage some students to practice in correctional institutions as a career. PMID- 14744712 TI - Informed consent skills in internal medicine residency: how are residents taught, and what do they learn? AB - PURPOSE: Obtaining informed consent is an essential skill in internal medicine (IM). The authors' informal observations and formal testing revealed deficiencies in residents' informed consent skills. This study evaluated how residents acquire informed consent skills and how informed consent skills are addressed in Canadian IM residency programs. METHOD: A questionnaire was delivered to all 16 IM program directors in Canada, asking how informed consent is taught and assessed. At the University of Saskatchewan IM residency program, residents were assessed through an objective structured clinical examination station, written examination, and a self-assessment questionnaire. RESULTS: No consistent approach to teaching or evaluating informed consent skills exists within Canadian IM programs. Program directors and residents identified informal mentoring by residents as an important learning modality. Although residents performed well in discussing procedural indications and techniques, discussing risks was inadequate. Residents focused on general and minor risks but avoided discussing serious risks and had difficulty discussing the frequency of complications. Residents lacked a structured approach to assessing capacity and often assessed only comprehension. Residents were unfamiliar with concepts such as material risk, implied consent, and therapeutic privilege. CONCLUSION: Explicit training in informed consent skills is urgently needed. Informal mentoring must be recognized as an important training method for informed consent and supported by appropriate teaching and evaluation strategies to ensure that resident-instructors do so effectively. PMID- 14744713 TI - Patient complaints about physician behaviors: a qualitative study. AB - PURPOSE: Health care institutions are required to routinely collect and address formal patient complaints. Despite the availability of this feedback, no published efforts explore such data to improve physician behavior. The authors sought to determine the usefulness of patient complaints by establishing meaningful categories and exploring their epidemiology. METHOD: A register of formal, unsolicited patient complaints collected routinely at the Wake Forest University Baptist Medical Center in Winston-Salem, North Carolina was used to categorize complaints using qualitative research strategies. After eliminating complaints unrelated to physician behavior, complaints from March 1999 were analyzed (60) to identify complaint categories that were then validated using complaints from January 2000 (122). Subsequently, all 1,746 complaints for the year 2000 were examined. Those unrelated to physician behavior (1,342) and with inadequate detail (182) were excluded, leaving 222 complaints further analysis. RESULTS: Complaints were most commonly lodged by patient (111), followed by a patient's spouse (33), (52), parent (50), relative/friend (15), or health professional (2). The most commonly identified category was disrespect (36%), followed by disagreement about expectations of care (23%), inadequate information (20%), distrust (18%), perceived unavailability (15%), interdisciplinary miscommunication (4%), and misinformation (4%). Multiple categories were identified in (19%) complaints. Examples from each category provide adequate detail to develop instructional modules. CONCLUSION: The seven complaint categories of physician behaviors should be useful in developing curricula related to professionalism, communication skills, practice-based learning. PMID- 14744714 TI - Do patients understand their physician's level of training? a survey of emergency department patients. AB - PURPOSE: To assess patients' understanding of levels of training and responsibilities for residents, medical students, and attendings in the emergency department as well as their degree of comfort in being cared for by a physician in-training. METHOD: In 1999, a questionnaire was administered to a convenience sample of 430 adult patients and family members in a university emergency department. The questionnaire asked for demographic information and contained 17 questions addressing the different levels of medical training and seven opinion based questions on patients' willingness to have physicians-in-training care for them. RESULTS: Respondents answered 65% of the knowledge-based questions about physicians' training correctly. Only 43% understood that residents are always supervised when caring for patients, and 30% thought attendings required supervision by a resident. Respondents with education greater than a high school diploma answered more questions correctly (71% versus 59%; p <.05). A total of 80% felt it was very important to know their physician's level of training, but only 58% reported actually knowing the level of training. Only 62% felt comfortable knowing that their physician might be a supervised physician-in training. In addition, despite the fact that this survey took place at a teaching hospital, 22% of respondents prefer not to be treated in a teaching hospital. CONCLUSIONS: Patients and their families do not fully understand the roles and responsibilities of the physicians-in-training that may be caring for them despite feeling it is important to know their physicians' level of training. PMID- 14744715 TI - Patients' willingness to allow residents to learn to practice medical procedures. AB - PURPOSE: Consent for teaching procedures has been the focus of ethical discussion recently. Patients may consent to a procedure but be unaware that the procedure is to be performed by a resident, perhaps for the first time. In such cases, patients have not specifically consented to the practice of teaching medical procedures. The authors studied patients' awareness of resident training and willingness to allow residents to perform procedures on them. METHOD: A survey was administered to a convenience sample of 202 Vanderbilt University Medical Center emergency department patients from February to April 2000. Three procedures (intubation, lumbar puncture, and sutures) were demonstrated. Patients were asked about their awareness of residents' training and willingness to allow a resident to perform the procedures for the first time versus the tenth time. RESULTS: In all, 60% of patients did not realize they could be the first person a resident performs a procedure on. Only 49% of the patients were completely comfortable being the first patient for sutures, 29% for intubation, and 15% for a lumbar puncture. Most patients felt they should be informed if it was the resident's first time performing procedures (66% for sutures, 69% for intubation, and 82% for lumbar puncture). CONCLUSION: Not only do the majority of patients not know that they might be the first patient on whom a resident performs a procedure, more than two thirds believed they should be told if they are the first patient. Particularly for intubation and lumbar puncture, patients indicated that they would be uncomfortable being the first patient on whom these procedures were performed. These data raise ethical questions regarding physicians' obligations to inform patients about resident-performed procedures. PMID- 14744716 TI - Doctors and patients: gender interaction in the consultation. AB - BACKGROUND: Much research on gender differences in medicine has centered on women as better communicators, more egalitarian, more patient-centered, and more involved with psychosocial problems, preventive care, and female-specific problems. Hardly any research has examined the interaction between the doctor's gender and the patient's gender. The authors examined students' perceptions and comfort levels regarding patients' gender during consultation. METHOD: This cross sectional study used a questionnaire to survey final-year medical students at one school in 1999. It tested students' patient-centeredness, "patient-care" values, and degree of comfort in performing certain intimate physical examinations. RESULTS: Women students were more patient-centered than were men students. Both genders were more attuned to the concerns of patients of their own gender, were more comfortable with personal rather than sexual issues, and were more uncomfortable with performing more intimate examinations upon the opposite gender. Using comparable case studies, it was also shown that the female student female patient dyad had significantly greater "patient-care" values than did the male student-male patient dyad. CONCLUSION: Medical students did not behave in a gender-neutral way in the consultation. There is a powerful interaction between a student's gender and a patient's gender. This warrants further investigation in the real clinical situation because it has implications on the outcomes of the consultation. PMID- 14744717 TI - Physicians' weight loss counseling in two public hospital primary care clinics. AB - PURPOSE: Primary care physicians are an important source of information on weight management. Nevertheless, weight loss counseling by these physicians remains inadequate. This study sought to determine physicians' barriers to providing weight loss counseling in a public hospital, patients' recall of physicians' weight loss recommendations, and the influence of physicians' counseling on patients' understanding, motivation, and behavior regarding weight loss. METHOD: In 2001, four focus groups of faculty and residents were held at two primary care clinics affiliated with the Louisiana State University Health Sciences Center Shreveport to determine the barriers to providing weight loss counseling. Scripted probes were used to uncover consensus norms. In 2001-02, structured exit interviews were conducted with 210 overweight or obese patients recruited from the clinics to determine patients' recall of physicians' weight loss recommendations, and patients' understanding of the relationship between weight and health, and their stages of readiness for weight loss. RESULTS: Physicians identified major barriers to providing weight loss counseling, including insufficient confidence, knowledge, and skills. Obesity was underdocumented as a distinct clinical diagnosis. Only 5% of the patients recalled being given the combined weight loss strategy of diet and exercise. However, patients who recalled being counseled to lose weight were more likely to understand the risks of obesity, the benefits of weight loss, and were at a higher stage of readiness for weight loss. CONCLUSIONS: Physicians' weight loss counseling had a significant effect on patients' understanding of and motivation for weight loss. However, physicians provided insufficient guidance on weight management strategies, possibly because of inadequate counseling skills and confidence. PMID- 14744718 TI - Behavioral coding for evaluation of medical student communication: clarification or obfuscation? AB - PURPOSE: To understand which specific student behaviors predict performance ratings from standardized patients and behavioral scientist preceptors. METHOD: In 1996-98, objective, real-time ratings of student verbal and nonverbal behaviors were conducted on 75 videotaped interviews between second-year medical students and standardized patients at University of North Carolina at Chapel Hill School of Medicine. The coding system used in these analyses was developed based on evidence-based literature reviews and used software that provides for real time recording; 30 nonverbal and 33 behaviors were coded. The coded behaviors were then compared with four ratings: a global rating of the encounter by the standardized patient, a summary score derived from a checklist completed by the standardized patient, a global rating of the encounter by a behavioral science preceptor who observed the encounter, and a summary score derived from a checklist completed by the preceptor. RESULTS: Analyses identified strong correlations between all four of the preceptor and patient ratings, a strong independent effect of case scenario, and significant between-rater variation. When multivariable analysis was used to predict these global ratings based on coding of specific behaviors, a relatively high proportion of observer variation was explained by a small group of coded behaviors. CONCLUSIONS: This study suggests that the coding of specific behaviors may hold promise as a student evaluation technique thereby improving medical training techniques and ultimately enhancing the communication skills in physician-patient encounters. It also illustrates the need to better clarify which specific behaviors are most critical in influencing patient satisfaction. PMID- 14744719 TI - A revisionist view of the integrated academic health center. AB - Like many academic health centers that had expanded aggressively during the 1990s, the nation's first vertically integrated academic health center, the University of Pennsylvania Health System, was profoundly challenged by the dramatic and unanticipated financial impacts of the Balanced Budget Act of 1997. The author explains why-although Penn's Health System had lost $300 million over two years and its debts threatened to cause serious financial and educational damage to the rest of the University-Penn chose to manage its way out of the financial crisis (instead of selling or spinning off its four hospitals, clinical practices, and possibly even its medical school). A strategy of comprehensive integration has not only stabilized Penn's Health System financially, but strengthened its position of leadership in medical education, research, and health care delivery. The author argues that a strategy of greater horizontal integration offers important strategic advantages to academic health centers. In an era when major social and scientific problems demand broadly multidisciplinary and highly-integrated approaches, such horizontally integrated institutions will be better able to educate citizens and train physicians, develop new approaches to health care policy, and answer pressing biomedical research questions. Institutional cultural integration is also crucial to create new, innovative organizational structures that bridge traditional disciplinary, school, and clinical boundaries. PMID- 14744720 TI - Teaching about the changing U.S. health care system: an innovative clerkship. AB - Changes in the U.S. health care system have necessitated modifying the scope and content of existing courses in the medical school curricula. In 1996, the Weill Medical College of Cornell University created a new, integrated public health curriculum to reflect the changes in the ways that medical care is organized, financed, and delivered. Teaching medical students to understand the constantly changing health care system is a primary objective of the new curriculum. As part of this curriculum, the medical college instituted a required public health clerkship that focused on the health care system, to be taken in either the third or fourth year. Students are prepared for the clerkship by taking courses in epidemiology, biostatistics, and evidence-based medicine in the first year and an introduction to the health system in the second year. The two-week clerkship, which may be unique in U.S. medical education, seeks to present an in-depth exposure to issues in health care financing and delivery by means of lectures, panel discussions with experts in the field, seminars, and field assignments to health care organizations and agencies. PMID- 14744723 TI - Night owl. Commentary. PMID- 14744724 TI - Communication failures: an insidious contributor to medical mishaps. AB - PURPOSE: To describe how communication failures contribute to many medical mishaps. METHOD: In late 1999, a sample of 26 residents stratified by medical specialty, year of residency, and gender was randomly selected from a population of 85 residents at a 600-bed U.S. teaching hospital. The study design involved semistructured face-to-face interviews with the residents about their routine work environments and activities, the medical mishaps in which they recently had been involved, and a description of both the individual and organizational contributory factors. The themes reported here emerged from inductive analyses of the data. RESULTS: Residents reported a total of 70 mishap incidents. Aspects of "communication" and "patient management" were the two most commonly cited contributing factors. Residents described themselves as embedded in a complex network of relationships, playing a pivotal role in patient management vis-a-vis other medical staff and health care providers from within the hospital and from the community. Recurring patterns of communication difficulties occur within these relationships and appear to be associated with the occurrence of medical mishaps. CONCLUSION: The occurrence of everyday medical mishaps in this study is associated with faulty communication; but, poor communication is not simply the result of poor transmission or exchange of information. Communication failures are far more complex and relate to hierarchical differences, concerns with upward influence, conflicting roles and role ambiguity, and interpersonal power and conflict. A clearer understanding of these dynamics highlights possibilities for appropriate interventions in medical education and in health care organizations aimed at improving patient safety. PMID- 14744726 TI - Helicobacter pylori eradication and gastric preneoplastic conditions: a randomized, double-blind, placebo-controlled trial. AB - Helicobacter pylori causes gastric adenocarcinoma; whether treatment of H. pylori infection prevents this cancer remains unknown. In a randomized, double-blind, placebo-controlled trial of H. pylori eradication, we determined whether treatment for H. pylori decreases gastric cancer risk, using preneoplastic conditions as surrogate markers. A total of 248 healthy volunteers (age >40 years) randomly received H. pylori treatment (omeprazole, amoxicillin, clarythromycin; n = 122) or matched placebo (n = 126) for 1 week. Endoscopy was performed at baseline and at 6 weeks and 1 year. Seven biopsies from each endoscopy were reviewed by two pathologists using the revised Sydney classification. Outcome measures were both a consensus "worst biopsy" diagnosis and a weighted index score that incorporated degrees of severity of preneoplasia from all biopsies. We compared change in these outcomes over time between the two treatment groups. H. pylori cure rates for compliant subjects in the treatment arm were 79.2% and 75.7% at 6 weeks and 1 year, respectively. No statistically significant change in the worst biopsy diagnosis was observed from 6 weeks to 1 year between placebo and treated subjects (for improvement/worsening, placebo, 19.4%/10.5%; treatment, 22.5%/8.3%; P = 0.74). Change in index score was favorably greater in treatment compared with placebo subjects (intention-to-treat analysis, P = 0.03); this finding was particularly evident in the antrum. H. pylori eradication gave more favorable gastric histopathologies over 1 year than no treatment. Such incomplete regression suggests but does not prove that eradication of H. pylori decreases cancer risk. PMID- 14744727 TI - p53 codon 72 polymorphism and cervical neoplasia: a meta-analysis review. AB - The Arg/Arg genotype versus Arg/Pro or Pro/Pro at codon 72 of the p53 gene has been implicated as a risk marker in cervical neoplasia. However, research on this topic has produced controversial results. We reviewed the published literature to summarize the association and to identify methodological features that may have contributed to the heterogeneity. Information on specific methodological features of studies addressing this topic published between 1998 and 2002 were obtained. Study-specific odds ratios (ORs) were combined in a meta-analysis, assuming random effects. To identify characteristics that significantly contributed to heterogeneity, we used meta-regression analysis. We identified 50 articles, of which 45 were included in the meta-analyses and regressions. No evidence of association or heterogeneity was detected for preinvasive lesions. For invasive cervical cancer with undefined histology, the Arg/Arg genotype was not found to affect risk (OR, 1.1; 95% confidence interval (CI), 0.9-1.3). However, a slightly increased risk was observed for squamous cell carcinoma (OR, 1.5; 95% CI, 1.2 1.9) and adenocarcinoma (OR, 1.7; 95% CI, 1.0-2.7). Meta-regression analysis identified that the most important factor contributing to heterogeneity among results for invasive lesions was departures from Hardy-Weinberg equilibrium in the control group. Summary ORs for studies in equilibrium were essentially null. A possible susceptibility role by the p53 codon 72 polymorphism at a late carcinogenetic stage in cervical cancer cannot be ruled out. However, various methodological features can contribute to departures from Hardy-Weinberg equilibrium and consequently to less than ideal circumstances for the examination of this polymorphism. Future investigations require appropriate attention to design and methodological issues. PMID- 14744728 TI - DNA repair gene XRCC1 and XPD polymorphisms and risk of prostate cancer. AB - The X-ray repair cross-complementing group 1 (XRCC1) and xeroderma pigmentosum group D (XPD) genes are involved in base excision repair and nucleotide excision repair of DNA repair pathways, respectively. A growing body of evidence suggests that XRCC1 and XPD are important in environmentally induced cancers, and polymorphisms in both genes have been identified. To determine whether the XRCC1 (codon Arg399Gln) and XPD (codon Asp312Asn and codon Lys751Gln) polymorphisms are associated with prostate cancer susceptibility, we genotyped these polymorphisms in a primarily Caucasian sample of 506 sibships (n = 1,117) ascertained through a brother with prostate cancer. Sibships were analyzed with a Cox proportional hazards model with age at prostate cancer diagnosis as the outcome. Of the three polymorphisms investigated, only the XPD codon 312 Asn/Asn genotype had an odds ratio (OR) significantly different from one (OR, 1.61; 95% CI, 1.03-2.53). Analyses stratified by the clinical characteristics of affected brothers in the sibship did not reveal any significant heterogeneity in risk. In exploring two way gene interactions, we found a markedly elevated risk for the combination of the XPD codon 312 Asn/Asn and XRCC1 codon 399 Gln/Gln genotypes (OR, 4.81; 95% CI, 1.66-13.97). In summary, our results suggest that the XPD codon 312 Asn allele may exert a modest positive effect on prostate cancer risk when two copies of the allele are present, and this effect is enhanced by the XRCC codon 399 Gln allele in its recessive state. PMID- 14744729 TI - Expression of the high mobility group proteins HMGI(Y) correlates with malignant progression in Barrett's metaplasia. AB - Expression of the high mobility group proteins HMGI(Y) has been shown to be a marker of malignancy in thyroid and pancreatic lesions and to correlate significantly with malignant progression in the colon. The aim of this study was to determine whether HMGI(Y) expression is associated with malignant progression in Barrett's metaplasia (BM). Immunoperoxidase staining for HMGI(Y) was performed on sections of formalin-fixed paraffin-embedded endoscopic esophageal biopsies from 42 patients with BM. These consisted of 19 biopsies negative for dysplasia (ND), 16 with low-grade dysplasia (LGD)/indeterminate for dysplasia (IND), and 7 with high-grade dysplasia (HGD)/adenocarcinoma (CA). The percentage of positive cells was recorded, and nuclear HMGI(Y) immunoreactivity in >10% of the cells was considered positive. Statistical analysis was performed using Fisher's exact test. Positive HMGI(Y) staining was detected in 2 of 19 (11%) cases ND, 5 of 16 (30%) LGD/IND cases, and 7 of 7 (100%) HGD/CA cases. Biopsies with HGD/CA were significantly more likely to be positive for HMGI(Y) than biopsies ND (P < 0.0001) or with LGD/IND (P = 0.0046). We conclude that HMGI(Y) expression is significantly associated with malignant progression in BM. Additional studies are needed to determine whether BM biopsies that are ND or LGD/IND and positive for HMGI(Y) are more likely to progress to adenocarcinoma. PMID- 14744730 TI - Nonsteroidal anti-inflammatory drug use associated with reduced incidence of adenocarcinomas of the esophagus and gastric cardia that overexpress cyclin D1: a population-based study. AB - This study was undertaken to determine whether selected risk factors for esophageal and gastric cancer are associated with tumors that overexpress cyclin D1. Archived tumor tissue was available for 630 esophageal and gastric cancer patients who participated in a population-based case-control study. Patients were categorized into case groups based on whether protein overexpression of the cyclin D1 gene, as assessed by immunohistochemistry, was present (cyclin D1+, n = 285) or not (cyclin D1-, n = 345) in the tumor. The distribution of risk factors in each of these case groups was then compared with the distribution among the 695 controls. Multivariate-adjusted odds ratios (OR) for esophageal adenocarcinoma were reduced in relation to use of aspirin and other nonsteroidal anti-inflammatory drug (NSAID) use but only among patients with cyclin D1+ tumors (0.45, 95% confidence interval [CI] = 0.26, 0.79) and not among those with cyclin D1- tumors (1.12, 95% CI = 0.67, 1.86). A similar pattern was observed for gastric cardia adenocarcinomas. In contrast, ORs for esophageal squamous cell carcinoma and noncardia gastric adenocarcinomas in relation to NSAID use were reduced, regardless of cyclin D1 status. ORs did not vary with cyclin D1 status in relation to alcohol, body size, or cigarette smoking, with the following exception; for noncardia gastric adenocarcinomas the cyclin D1- tumors showed a 2 fold elevation in the OR with ever smoking. These data suggest that the reduction in risk associated with NSAID use may be restricted to those esophageal and gastric cardia adenocarcinomas that overexpress cyclin D1. PMID- 14744731 TI - Dietary carotenoids and risk of lung cancer in a pooled analysis of seven cohort studies. AB - Intervention trials with supplemental beta-carotene have observed either no effect or a harmful effect on lung cancer risk. Because food composition databases for specific carotenoids have only become available recently, epidemiological evidence relating usual dietary levels of these carotenoids with lung cancer risk is limited. We analyzed the association between lung cancer risk and intakes of specific carotenoids using the primary data from seven cohort studies in North America and Europe. Carotenoid intakes were estimated from dietary questionnaires administered at baseline in each study. We calculated study-specific multivariate relative risks (RRs) and combined these using a random-effects model. The multivariate models included smoking history and other potential risk factors. During follow-up of up to 7-16 years across studies, 3,155 incident lung cancer cases were diagnosed among 399,765 participants. beta Carotene intake was not associated with lung cancer risk (pooled multivariate RR = 0.98; 95% confidence interval, 0.87-1.11; highest versus lowest quintile). The RRs for alpha-carotene, lutein/zeaxanthin, and lycopene were also close to unity. beta-Cryptoxanthin intake was inversely associated with lung cancer risk (RR = 0.76; 95% confidence interval, 0.67-0.86; highest versus lowest quintile). These results did not change after adjustment for intakes of vitamin C (with or without supplements), folate (with or without supplements), and other carotenoids and multivitamin use. The associations generally were similar among never, past, or current smokers and by histological type. Although smoking is the strongest risk factor for lung cancer, greater intake of foods high in beta-cryptoxanthin, such as citrus fruit, may modestly lower the risk. PMID- 14744732 TI - Cigarette smoking and testicular cancer. AB - The purpose of this study was to determine the relation between testicular cancer and cigarette smoking. Data were collected between 1995 and 1996 in Ontario, Canada, as part of the Enhanced Cancer Surveillance Study. Pack-years and years of smoking were examined among all subjects (212 cases and 252 controls) and former and current smokers. Years since quitting and age at smoking initiation were examined among former and current smokers only. Independent of smoking status, significant associations were noted among those who smoked between 12 and 24 pack-years [odds ratio (OR) = 1.96 (95% confidence interval (CI): 1.04-3.69), relative to nonsmokers] or greater [>24 pack-years, OR = 2.31 (95% CI: 1.12 4.77), relative to nonsmokers], and among those who smoked >21 years [OR = 3.18 (95% CI: 1.32-7.64), relative to nonsmokers]. Quitting smoking was not found to result in a reduction of risk. No association was observed for smoking at adolescence relative to a later period. Results from the study suggest that cigarette smoking exerts an adverse influence on testicular cancer risk that is not mitigated by smoking cessation and not altered by age at initiation. PMID- 14744733 TI - Smokeless and other noncigarette tobacco use and pancreatic cancer: a case control study based on direct interviews. AB - Cigarette smoking is an important and well-established cause of pancreatic cancer. In contrast, little is known about the effects of smoking cigars, pipes, and use of smokeless tobacco on pancreatic cancer risk. The objective of the present study was to examine the association between noncigarette tobacco use (i.e., cigars, pipes, smokeless tobacco) and pancreatic cancer risk among nonsmokers of cigarettes. A population-based case-control study of pancreatic cancer was conducted during 1986-1989 among residents of Atlanta, Georgia, Detroit, Michigan, and 10 counties in New Jersey. Direct interviews were successfully completed with 526 newly diagnosed pancreatic cancer patients and 2153 controls ages 30-79 years. This analysis was restricted to lifelong nonsmokers of cigarettes and based on interviews with 154 cases newly diagnosed with carcinoma of the exocrine pancreas and 844 population controls who reported no history of cigarette smoking. We observed a consistent pattern of increased risk associated with cigar smoking, although these elevations were not statistically significant. Participants who smoked cigars regularly (i.e., at least one cigar/week for >/=6 months) experienced a 70% increased risk [95% confidence interval (CI): 0.9-3.3], and those who never used other form of tobacco had a 90% increased risk (95% CI: 0.8-4.3). Risk was elevated among those who smoked more than one cigar/day [odds ratio (OR) = 1.8; 95% CI: 0.8-4.2) and among those who smoked cigars > 20 years (OR = 1.9; 95% CI: 0.9-3.9). Trends in risk with increasing amount and duration smoked were consistent but not statistically significant (P = 0.17 and P = 0.16, respectively). Subjects who used smokeless tobacco regularly had a 40% increased risk of pancreatic cancer (95% CI: 0.5-3.6) compared with nonusers of tobacco. We observed a marginally significant increasing risk with increased use of smokeless tobacco (P = 0.04); participants who used >2.5 oz of smokeless tobacco a week had an OR of 3.5 (95% CI: 1.1-11). Long-term use of smokeless tobacco (i.e., >20 years) was also associated with a nonsignificant increased risk (OR = 1.5; 95% CI: 0.6-4.0). In contrast, pipe smokers experienced no increased risk (OR = 0.6; 95% CI: 0.1-2.8). Our results suggest that heavy use of smokeless tobacco, and to a lesser extent, cigar smoking may increase the risk of pancreatic cancer among nonsmokers of cigarettes. PMID- 14744734 TI - Ultraviolet radiation and incidence of non-Hodgkin's lymphoma among Hispanics in the United States. AB - Non-Hodgkin's lymphoma (NHL) is one of the most common cancers among American Hispanics. Several studies suggest that solar UV radiation (UVR) may be an environmental risk contributing to the rise of NHL over the past decades. These studies focused primarily on light-skinned Caucasian populations; it is unknown what role UVR plays in NHL for Hispanics. We described the incidence of NHL in Hispanics from selected states in the United States between 1989 and 2000. To evaluate the role of UVR, we correlated cancer rates with the UV index and latitude of residency. Variations in NHL incidence rates with estimated amount of UVR among whites and blacks from the selected states were also analyzed. We found that NHL occurred less frequently in Hispanics than in whites. Hispanic men had higher incidence of NHL than Hispanic women. Incidence rates of NHL in Hispanics were inversely associated with estimated amount of UVR as an increase in NHL was observed with decreasing UV index (r = -0.7 in men; r = -0.41 in women) or increasing latitude of residency (r = 0.59 in men; r = 0.48 in women). This trend, although it did not reach statistical significance, was also observed in whites and blacks. Our findings do not support previous reports of a positive association between solar radiation and NHL. The inverse relationship between UVR and incidence of NHL is unexplained but presents the need for generation of hypotheses regarding the epidemiology of causal factors for NHL in the United States. Additional studies should be conducted to clarify whether sunlight exposure contributes to the development of NHL. PMID- 14744736 TI - Dietary patterns and prostate cancer risk in the National Health and Nutrition Examination Survey Epidemiological Follow-up Study cohort. AB - Ecological studies implicate a "Western" diet in prostate cancer development, but whether dietary patterns measured in individuals are associated with risk has not been studied previously. We examined this issue using prospective data from the nationally representative United States Health Examination Epidemiological Follow up Study. Among 3,779 men followed from 1982-84 to 1992, 136 incident cases were identified. Using principal component analysis on responses to a 105-item dietary questionnaire, the following three distinct patterns were identified: a vegetable fruit pattern; a red meat-starch pattern characterized by red meats, potatoes, cheese, salty snacks, and desserts; and a Southern pattern characterized by such foods as cornbread, grits, sweet potatoes, okra, beans, and rice. In adjusted proportional hazards models, prostate cancer risk was not associated with the vegetable-fruit or red meat-starch pattern, but higher intake of the Southern pattern showed a reduction in risk (3rd versus 1st tertile relative risk, 0.6; 95% confidence interval, 0.4-1.1; trend P = 0.08) that approached statistical significance. The inverse association was observed in black and non-black men and was not attributable to intake of any individual foods or nutrients. A Southern dietary pattern may reflect a history of living in the South and serve as an integrative marker of sunlight exposure and protection through 1,25 dihydroxyvitamin D production. Further evaluation and better characterization of the pattern would offer more information on potentially beneficial features of the diet or its associated lifestyle. PMID- 14744735 TI - Dietary glycemic load and breast cancer risk in the Women's Health Study. AB - A diet with a high glycemic load (GL) may contribute to a metabolic environment that enhances tumorigenesis. Little is known, however, about whether high glycemic diets increase breast cancer risk in women. We examined the associations between baseline measurements of dietary GL and overall glycemic index (GI) and subsequent breast cancer in a cohort of 39,876 women, ages 45 years or older, participating in the Women's Health Study. During a mean of 6.8 years of follow up there were 946 confirmed cases of breast cancer. We found no association between dietary GL [multivariable-adjusted relative risk (RR), 1.01; confidence interval (CI), 0.76-1.35, comparing extreme quintiles; P for trend = 0.96] or overall GI (corresponding RR, 1.03; CI, 0.84-1.28; P for trend = 0.66) and breast cancer risk in the cohort as a whole. Exploratory analyses stratified by baseline measurements of menopausal status, physical activity, smoking history, alcohol use, and history of diabetes mellitus, hypertension, or hypercholesterolemia showed no significant associations, except in the subgroup of women who were premenopausal and reported low levels of physical activity (GL multivariable adjusted RR, 2.35; CI, 1.03-5.37; P for trend = 0.07; GI multivariable-adjusted RR, 1.56; CI, 0.88-2.78; P for trend = 0.02, comparing extreme quintiles). Although we did not find evidence that a high glycemic diet increases overall breast cancer risk, the increase in risk in premenopausal women with low levels of physical activity suggests the possibility that the effects of a high glycemic diet may be modified by lifestyle and hormonal factors. Prospective studies of a larger sample size and longer duration are warranted to confirm our findings. PMID- 14744737 TI - Behaviors used by men to protect themselves against prostate cancer. AB - This paper reports on behaviors men use to protect themselves against prostate cancer. Data were collected via a telephone or mailed survey from 353 men enrolled in two studies of prostate cancer screening. Respondents reported behaviors they used to protect themselves against prostate cancer, and responses were coded as conventional care, self-care, or nothing. Men who reported using both conventional care and self-care were categorized as conventional care users. Polytomous logistic regression was conducted to evaluate the association between sociodemographic background, prior prostate screening, and cognitive, affective, and social support and influence factors with protective behavior type. The distribution of protective behaviors was as follows: conventional care, 63%; self care only, 19%; and nothing, 18%. In multivariable analyses, higher education level was found to be positively associated with conventional care use. Perceived salience and coherence of prostate cancer screening was positively associated with conventional care use among men in one of the two studies. Low concern about screening was positively associated with self-care use, as was mailed survey completion. This study presents self-report data regarding prostate cancer protection behaviors. Most men in the study reported using some type of prostate cancer protective behavior. Decision-making about whether or not to take protective action and what type of behavior to use may be influenced by socioeconomic background, cognitive perceptions related to behavioral options, and concern about risk. PMID- 14744738 TI - Risk estimation for healthy women from breast cancer families: new insights and new strategies. AB - Risk estimation in breast cancer families is often estimated by use of the Claus tables. We analyzed the family histories of 196 counselees; compared the Claus tables with the Claus, the BRCA1/2, the BRCA1/2/ models; and performed linear regression analysis to extend the Claus tables with characteristics of hereditary breast cancer. Finally, we compared the Claus extended method with the Claus, the BRCA1/2, and the BRCA1/2/u models. We found 47% agreement for Claus table versus Claus model; 39% agreement for Claus table versus BRCA1/2 model; 48% agreement for Claus table versus BRCA1/2/u model; 37% agreement for Claus extended method versus Claus model; 44% agreement for Claus extended model versus BRCA1/2 model; and 66% agreement for Claus extended method versus BRCA1/2/u model. The regression formula (Claus extended method) for the lifetime risk for breast cancer was 0.08 + 0.40 (*) Claus Table + 0.07 (*) ovarian cancer + 0.08 (*) bilateral breast cancer + 0.07 (*) multiple cases. This new method for risk estimation, which is an extension of the Claus tables, incorporates information on the presence of ovarian cancer, bilateral breast cancer, and whether there are more than two affected relatives with breast cancer. This extension might offer a good alternative for breast cancer risk estimation in clinical practice. PMID- 14744739 TI - Association of CYP17, CYP19, CYP1B1, and COMT polymorphisms with serum and urinary sex hormone concentrations in postmenopausal women. AB - Women with high circulating estrogen concentrations have an increased risk of breast cancer; thus, it is important to understand factors, including genetic variability, that influence estrogen concentrations. Several genetic polymorphisms that may influence sex hormone concentrations have been identified, including CYP17 (5'-untranslated region T-->C), CYP19 [intron 4 (TTTA)(n = 7-13) and a 3-bp deletion (-3)], CYP1B1 (Val(432)Leu), and COMT (Val(108/158)Met). We examined associations between these polymorphisms and serum concentrations of estrogens, androgens, and sex hormone-binding globulin and urinary concentrations of 2- and 16alpha-hydroxyestrone in 171 postmenopausal women, using data from the prerandomization visit of an exercise clinical trial. Participants were sedentary, not taking hormone therapy, and had a body mass index >24.0. Compared with noncarriers, women carrying two CYP19 7r(-3) alleles had 26% lower estrone (P < 0.001), 19% lower estradiol (P = 0.01), 23% lower free estradiol (P = 0.01), and 22% higher sex hormone-binding globulin concentrations (P = 0.06). Compared with noncarriers, women carrying at least one CYP19 8r allele had 20% higher estrone (P = 0.003), 18% higher estradiol (P = 0.02), and 21% higher free estradiol concentrations (P = 0.01). Women with the COMT Met/Met genotype had 28% higher 2-hydroxyestrone (P = 0.08) and 31% higher 16alpha-hydroxyestrone concentrations (P = 0.02), compared with Val/Val women. Few associations were found for CYP17 and CYP1B1 or with serum androgen concentrations. This study provides further evidence that genetic variation may appreciably alter sex hormone concentrations in postmenopausal women not taking hormone therapy. PMID- 14744740 TI - Correlation between the UDP-glucuronosyltransferase (UGT1A1) TATAA box polymorphism and carcinogen detoxification phenotype: significantly decreased glucuronidating activity against benzo(a)pyrene-7,8-dihydrodiol(-) in liver microsomes from subjects with the UGT1A1*28 variant. AB - Of the hepatic UDP-glucuronosyltransferases (UGTs), only UGT1A1 and UGT1A9 exhibit activity against benzo(a)pyrene-trans-7R,8R-dihydrodiol [BPD(-)], precursor to the highly mutagenic anti-(+)-benzo(a)pyrene-7R,8S-dihydrodiol 9S,10R-epoxide. The UGT1A1*28 allelic variant contains an additional (TA) dinucleotide repeat in the "TATAA" box [(TA)(6)>(TA)(7)] of the UGT1A1 promoter that has been linked to decreased expression of the UGT1A1 gene and decreased bilirubin conjugation, leading to the relatively nondebilitating condition known as Gilbert's syndrome. To determine whether the UGT1A1 TATAA box polymorphism may play a role in the overall glucuronidation of BPD(-) in humans, we compared UGT1A1 TATAA box genotype with BPD(-) glucuronidating activity in normal liver microsomes. Significant decreases in UGT1A1 protein (P < 0.005) and bilirubin conjugation activity (P < 0.001) were observed in liver microsomes from subjects homozygous for the UGT1A1*28 allelic variant compared with subjects homozygous for the wild-type UGT1A1*1 allele. Significant decreases in BPD(-) glucuronidation activity (P < 0.02) were observed in subjects with the UGT1A1(*28/*28) genotype compared with subjects having the wild-type UGT1A1(*1/*1) genotype in assays of liver microsomes that included 0.1 mM alpha naphthylamine, a competitive inhibitor of UGT1A9 and not UGT1A1. Similar phenotype:genotype correlations were observed when we compared subjects with the UGT1A1(*28/*28) genotype with subjects having the UGT1A1(*1/*28) genotype. In assays with alpha-naphthylamine, the K(m) of liver microsomes against BPD(-) was similar to that reported for UGT1A1-overexpressing baculosomes (319 micro M versus 290 micro M; Fang et al., Cancer Res., 62: 1978-1986, 2002). These data suggest that the UGT1A1 TATAA box polymorphism plays a role in an individual's overall ability to detoxify benzo(a)pyrene and in cancer risk. PMID- 14744741 TI - Natural history of human papillomavirus type 16 virus-like particle antibodies in young women. AB - Immunization with a vaccine of human papillomavirus (HPV) type 16 virus-like particles (VLPs) can reduce incidence of HPV-16 infection and its related cervical intraepithelial neoplasia. However, development of detectable antibodies to VLPs does not always occur after natural HPV infection. This study examined prospectively for seroconversion and duration of antibodies to HPV-16 VLPs and their associated host and viral factors. Six-hundred eight subjects were tested for HPV DNA biannually and for IgG and IgA antibodies to HPV-16 VLPs annually for 3 years. Both IgG and IgA antibodies to HPV-16 VLPs were predominantly type specific. Women with cervicovaginal HPV-16 infection were 8-10 times more likely to seroconvert than those with infection of HPV-16-related types. Among subjects who had an incident infection with HPV-16, a maximum of 56.7% became seropositive for IgG within 8.3 months and 37.0% had IgA within 14 months. Detectable seroconversion was a slow process that required sufficient antigenic exposure associated with either a high viral load (relative risk = 5.7 for IgG) or persistent infection of HPV-16 (relative risk = 3.4 for IgA). The median duration for both types of antibodies was approximately 36 months. Antibodies could persist for a long period of time if the initial antibody levels were high or if there was continued antigenic exposure. PMID- 14744742 TI - Albumin adducts of naphthalene metabolites as biomarkers of exposure to polycyclic aromatic hydrocarbons. AB - We investigated the utility of adducts formed by the reaction of the naphthalene metabolites naphthalene-1,2-oxide, 1,2-naphthoquinone (1,2-NPQ), and 1,4 naphthoquinone (1,4-NPQ) with serum albumin (Alb) as biomarkers of exposure to polycyclic aromatic hydrocarbons. Cysteinyl serum Alb adducts of 1,2- and 1,4-NPQ (1,2-NPQ-Alb and 1,4-NPQ-Alb, respectively) but not of naphthalene-1,2-oxide were detected in 28 coke oven workers and 22 controls from the steel industry of northern China. The median level of 1,2-NPQ-Alb in coke oven workers (76.6 pmol/g) was significantly higher than that observed in controls (44.9 pmol/g; P = 0.0027). However, the median level of 1,4-NPQ-Alb in exposed subjects was not significantly different from that of controls (48.6 versus 44.2 pmol/g; P = 0.296). Levels of 1,2-NPQ-Alb were significantly correlated with exposure category (controls, side and bottom workers, and top-of-oven workers) as well as with previously measured levels of urinary naphthalene, 1- and 2-naphthol, and 1 pyrenol in these subjects. Multiple linear regression analysis revealed that 35% of the variation in 1,2-NPQ-Alb could be explained by the work category and age. A negative relationship between 1,2-NPQ-Alb and age was observed, suggesting that cytochrome P450 c metabolism diminished with age at approximately 3%/year of life. PMID- 14744743 TI - Hydrolysis of glucosinolates to isothiocyanates after ingestion of raw or microwaved cabbage by human volunteers. AB - Cabbage contains the glucosinolate sinigrin, which is hydrolyzed by myrosinase to allyl isothiocyanate. Isothiocyanates are thought to inhibit the development of cancer cells by a number of mechanisms. The effect of cooking cabbage on isothiocyanate production from glucosinolates during and after their ingestion was examined in human subjects. Each of 12 healthy human volunteers consumed three meals, at 48-h intervals, containing either raw cabbage, cooked cabbage, or mustard according to a cross-over design. At each meal, watercress juice, which is rich in phenethyl isothiocyanate, was also consumed to allow individual and temporal variation in postabsorptive isothiocyanate recovery to be measured. Volunteers recorded the time and volume of each urination for 24 h after each meal. Samples of each urination were analyzed for N-acetyl cysteine conjugates of isothiocyanates as a measure of entry of isothiocyanates into the peripheral circulation. Excretion of isothiocyanates was rapid and substantial after ingestion of mustard, a source of preformed allyl isothiocyanate. After raw cabbage consumption, allyl isothiocyanate was again rapidly excreted, although to a lesser extent than when mustard was consumed. On the cooked cabbage treatment, excretion of allyl isothiocyanate was considerably less than for raw cabbage, and the excretion was delayed. The results indicate that isothiocyanate production is more extensive after consumption of raw vegetables but that isothiocyanates still arise, albeit to a lesser degree, when cooked vegetables are consumed. The lag in excretion on the cooked cabbage treatment suggests that the colon microflora catalyze glucosinolate hydrolysis in this case. PMID- 14744744 TI - Delivery of tea polyphenols to the oral cavity by green tea leaves and black tea extract. AB - Catechins and theaflavins, polyphenolic compounds derived from tea (Camellia sinensis, fam. Theaceae), have been reported to have a wide range of biological activities including prevention of tooth decay and oral cancer. The present study was undertaken to determine the usefulness of green tea leaves and black tea extract for the delivery of catechins and theaflavins to the oral cavity. After holding either green tea leaves (2 g) or brewed black tea (2 g of black tea leaves in 100 ml) in the mouth for 2-5 min and thoroughly rinsing the mouth, high concentrations of catechins (C(max) = 131.0-2.2 micro M) and theaflavins (C(max) = 1.8-0.6 micro M) were observed in saliva in the 1st hour. Whereas there was significant interindividual variation in the peak levels of catechins and theaflavins, the overall kinetic profile was similar, with t(1/2) = 25-44 min and 49-76 min for catechins and theaflavins, respectively (average coefficient of variation in t(1/2) was 23.4%). In addition to the parent catechin and theaflavin peaks, five unidentified peaks were also observed in saliva after black tea treatment. Hydrolysis of theaflavin gallates, apparently by salivary esterases, was observed in vitro and in vivo. These results indicate that tea leaves can be used as a convenient, slow-release source of catechins and theaflavins and provide information for the possible use of tea in the prevention of oral cancer and dental caries. PMID- 14744745 TI - Expert review of non-Hodgkin's lymphomas in a population-based cancer registry: reliability of diagnosis and subtype classifications. AB - Incidence rates of non-Hodgkin's lymphomas (NHLs) have nearly doubled in recent decades. Understanding the reasons behind these trends will require detailed surveillance and epidemiological study of NHL subtypes in large populations, using cancer registry or other multicenter data. However, little is known regarding the reliability of NHL diagnosis and subtype classification in such data, despite implications for the accuracy of incidence statistics and studies. Expert pathological re-review was completed for 1526 NHL patients who were reported to the Greater Bay Area Cancer Registry and who participated in a large population-based case-control study. Agreement of registry diagnosis with expert diagnosis and with International Classification of Diseases for Oncology-2 (Working Formulation) subtype classifications was measured with positive predictive values and kappa statistics. Agreement of registry and expert diagnoses was high (98%). Thirty patients were found on review not to have NHL; most of these had leukemia. For subtypes, agreement of registry and expert classification was more moderate (59%). Agreement varied substantially by subtype from 5% to 100% and was 77% for the most common subtype, diffuse large cell lymphoma. Seventy-seven percent of 128 registry-unclassified lymphomas were assigned a subtype on re-review. Our analyses suggest excellent diagnostic reliability but poorer subtype reliability of NHL in cancer registry data information that is critical to the interpretation of lymphoma time trends. Thus, overall NHL incidence and survival statistics from the early 1990s are probably accurate, but subtype-specific statistics could be substantially biased, especially because of high (15-20%) proportions of unclassified lymphomas. PMID- 14744746 TI - A mechanism for cox-2 inhibitor anti-inflammatory activity in chemoprevention of epithelial cancers. PMID- 14744747 TI - Genetic variants of GPX1 and SOD2 and breast cancer risk at the Ontario site of the Breast Cancer Family Registry. AB - There are several genes that code for enzymes, including various forms of superoxide dismutase and glutathione peroxidase, that protect the cell against oxidative damage that, in turn, can lead to carcinogenesis. There are a few common genetic polymorphisms in these genes that lead to altered proteins. Three that have been identified are SOD2 Val-9Ala, GPX1 Pro198Leu, and the GPX1 GCG repeat (three alleles with four, five, or six repeats). The SOD2 variant has been associated with increased breast cancer risk in two studies. The GPX1 variants have not been studied with respect to breast cancer, but Pro198Leu has been associated with lung cancer. We conducted a case-control study of these three polymorphisms in incident, invasive breast cancer in Caucasian women under 55. There were 399 cases and 372 controls genotyped, of whom 488 were premenopausal, 208 postmenopausal, and 75 of unknown menopausal status. We were unable to replicate the previously observed association with SOD2 Val-9Ala and also found no association between breast cancer and GPX1 Pro198Leu. However, the allele of GPX1 containing four GCG repeats was significantly associated with breast cancer risk in premenopausal women (odds ratio, 1.55; 95% confidence interval, 1.04-2.30 for carriers versus noncarriers). There is a significant trend of increasing risk with increasing number of alleles with four GCG repeats (P = 0.03). This variant has not previously been reported to be associated with breast cancer. PMID- 14744748 TI - Ataxia-telangiectasia-mutated (ATM) gene in head and neck squamous cell carcinoma: promoter hypermethylation with clinical correlation in 100 cases. AB - The Ataxia-telangiectasia-mutated (ATM) gene product is a well-characterized tumor suppressor that plays a key role in maintenance of genomic stability. We have recently documented that the ATM promoter is a target for epigenetic silencing in cultured tumor cells. Here we show that aberrant methylation of the ATM promoter occurs in a significant percentage (25%) of head and neck squamous cell carcinomas. The presence of methylated ATM promoter shows a statistically significant correlation with an earlier age of initial diagnosis and decreased overall survival, particularly in early-stage tumors. These findings indicate that ATM promoter hypermethylation occurs in head and neck squamous cell carcinoma, and this feature is a potentially useful prognostic marker in this tumor type. PMID- 14744749 TI - Methionine synthase D919G polymorphism, folate metabolism, and colorectal adenoma risk. AB - Methionine synthase [5-methyltetrahydrofolate-homocysteine S-methyltransferase (MTR)] is involved in folate-mediated one-carbon metabolism, a pathway known to play a role in colorectal carcinogenesis. We investigated whether the MTR D919G polymorphism was associated with risk of colorectal adenoma in a colonoscopy based study of 513 cases and 609 controls from Minneapolis, MN. Adenoma risk appeared nonsignificantly increased among women with DG or GG genotype [adjusted odds ratio (OR) versus DD, 1.4; 95% confidence interval (CI), 0.9-2.1] but not men (OR, 1.0; 95% CI, 0.7-1.5). An interaction with methionine intake was observed among women, such that low versus high intake was associated with a 2.3 fold increased risk only among those with DG or GG genotype (95% CI, 1.1-4.9; P for interaction = 0.05). Similarly, risk associated with alcohol intake was not elevated among women with the DD genotype; however, consumption of >7 g of alcohol/day versus none was associated with an increased risk among women with DG or GG genotype (adjusted OR, 2.5; 95% CI, 1.4-4.4; P for interaction = 0.03). An interaction between MTR D919G and the thymidylate synthase (TS or TYMS) 3' untranslated region polymorphism 1494del6 was also observed among women (P for interaction = 0.007). No evidence of interaction with intake of folate, vitamin B(12), or vitamin B(6) or with genotype at MTHFR C677T or the TS enhancer region 28-bp repeat polymorphism was seen. These findings add to what is known about the complexities of genetic variations in one-carbon-metabolizing enzymes in relation to colorectal carcinogenesis. PMID- 14744750 TI - Serum insulin-like growth factor-I is positively associated with serum prostate specific antigen in middle-aged men without evidence of prostate cancer. AB - We have examined the relationship between serum insulin-like growth factor-I (IGF I) and prostate-specific antigen in 367 healthy men without evidence of prostate cancer and found a positive association (P = 0.05). In men without prostate cancer, serum prostate-specific antigen is closely related to prostate size, and our findings, therefore, suggest that IGF-I may induce prostatic epithelial proliferation. Higher circulating levels of IGF-I have been associated with increased risk of both prostate cancer and possibly benign prostatic hyperplasia. Greater rates of cell proliferation induced by IGF-I may be a key biological pathway underlying these disorders. PMID- 14744751 TI - Association between insulin-like growth factor-I: insulin-like growth factor binding protein-1 ratio and metabolic and anthropometric factors in men and women. AB - Several prospective observational studies have suggested that elevated circulating IGF-I levels are associated with an increased risk of cancer. These observations may provide a potential mechanism through which previously identified metabolic and anthropometric factors, such as obesity and elevated insulin and glucose levels, may operate. We therefore examined metabolic and anthropometric influences on circulating levels of insulin-like growth factor-I (IGF-I), insulin-like growth factor-binding protein-1 (IGFBP-1), and the IGF I:IGFBP-1 ratio in a middle-aged population of 349 men and 492 women. IGF-I showed only modest inverse associations with indices of adiposity. However, we found that low IGFBP-I levels and an increased IGF-I:IGFBP-1 ratio were strongly associated with increased levels of insulin and glucose in men and women. Body mass index was also positively related to the IGF-I:IGFBP-1 ratio in men (P < 0.001) and women (P < 0.001), independent of metabolic correlates of IGFBP-1 and IGF-I. Similarly, waist:hip ratio and waist circumference were also associated with an increased IGF-I:IGFBP-1 ratio and low circulating IGFBP-1 levels. These findings suggest that individuals with greater fat mass and upper body obesity may have elevated levels of bioavailable or free IGF-I, which could, in part, mediate the reported associations among metabolic and anthropometric factors and cancer risk. PMID- 14744752 TI - Combinatorial screenings in patients: the interleukin-11 receptor alpha as a candidate target in the progression of human prostate cancer. AB - Direct screening of combinatorial peptide libraries in patients may allow the identification of ligands that target biochemical differences in the endothelium of blood vessels. In a screening performed in a patient, we selected and isolated a mimic motif of interleukin 11 (IL-11) from prostate biopsies after an i.v. administration of a phage display peptide library. We also demonstrated that the IL-11 phage mimic (displaying the cyclic nonapeptide CGRRAGGSC) bound specifically to a corresponding IL-11 receptor (IL-11Ralpha). Here we show that IL-11Ralpha is a potential target for intervention in human prostate cancer through morphological and functional analyses. First, a comprehensive serial immunohistochemical analysis of primary and metastatic prostate cancer samples showed increased stage-specific expression of IL-11Ralpha during disease progression. Second, a proapoptotic peptide was specifically targeted and internalized through this functional IL-11Ralpha-based ligand-receptor pair: treatment of prostate cancer cells in vitro with a proapoptotic peptide guided by the CGRRAGGSC peptide to the IL-11Ralpha resulted in dose-dependent apoptosis. Together, these data indicate that the IL-11Ralpha is a candidate target for translational clinical trials against advanced and metastatic prostate cancer. Moreover, our results illustrate the ability of direct combinatorial screening systems in cancer patients for identification of relevant targets in the context of human disease. PMID- 14744753 TI - Slippage of mitotic arrest and enhanced tumor development in mice with BubR1 haploinsufficiency. AB - A compromised spindle checkpoint is thought to play a key role in genetic instability that predisposes cells to malignant transformation. Loss of function mutations of BubR1, an important component of the spindle checkpoint, have been detected in human cancers. Here we show that BubR1(+/-) mouse embryonic fibroblasts are defective in spindle checkpoint activation, contain a significantly reduced amount of securin and Cdc20, and exhibit a greater level of micronuclei than do wild-type cells. RNA interference-mediated down-regulation of BubR1 also greatly reduced securin level. Moreover, compared with wild-type littermates, BubR1(+/-) mice rapidly develop lung as well as intestinal adenocarcinomas in response to challenge with carcinogen. BubR1 is thus essential for spindle checkpoint activation and tumor suppression. PMID- 14744754 TI - Tobacco carcinogen-induced cellular transformation increases activation of the phosphatidylinositol 3'-kinase/Akt pathway in vitro and in vivo. AB - The role of the phosphatidylinositol 3'-kinase (PI3K)/Akt pathway during tobacco carcinogen-induced transformation is unknown. To address this question, we evaluated this pathway in isogenic immortalized or tumorigenic human bronchial epithelial cells in vitro, as well as in progressive murine lung lesions induced by a tobacco-specific carcinogen, 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone. Compared with immortalized cells, tumorigenic cells had greater activation of the PI3K/Akt pathway, enhanced survival, and increased apoptosis in response to inhibition of the pathway. In vivo, increased activation of Akt and mammalian target of rapamycin was observed with increased phenotypic progression. Collectively, these results support the hypothesis that maintenance of Akt activity is necessary for survival of preneoplastic as well as transformed lung epithelial cells and suggest that inhibition of the PI3K/Akt pathway might be a useful approach to arrest lung tumorigenesis. PMID- 14744755 TI - T-bet regulates metastasis rate in a murine model of primary prostate cancer. AB - The local progression of primary tumors is extrinsically controlled by type 1 immune responses, particularly via the cytokine IFN-gamma, whose secretion is highly dependent on helper T cells. The T-box transcription factor T-bet (Tbx21) plays a critical role in the development of type 1 helper T cells and is essential for the production of IFN-gamma. Here, the T-bet pathway in the autochthonous transgenic adenocarcinoma mouse prostate model is demonstrated to have only a modest effect on the characteristics of primary prostate cancers but rather exerts a significant suppressor function in the development of metastatic disease. PMID- 14744756 TI - Protein kinase C zeta transactivates hypoxia-inducible factor alpha by promoting its association with p300 in renal cancer. AB - Hydroxylation at an asparagine residue at the COOH-terminal activation domain of hypoxia-inducible factor (HIF)-1/2 alphas is essential for its inactivation under normoxic condition. To date, the mechanism by which HIF-alpha avoids the inhibitory effect of asparagine hydroxylase in renal cell carcinoma (RCC) in normoxia is undefined. We have shown herein that protein kinase C (PKC) zeta has an important role in HIF-alpha activation in RCC. By using dominant negative mutant and small interference RNA approaches, we have demonstrated that the association between HIF-alpha and p300 is modulated by PKCzeta. Moreover, a novel signaling pathway involving phosphatidylinositol 3'-kinase and PKCzeta has been shown to be responsible for the activation of HIF-alpha by inhibiting the mRNA expression of FIH-1 (factor inhibiting HIF-1) in RCC and thereby promoting the transcription of hypoxia-inducible genes such as vascular permeability factor/vascular endothelial growth factor. PMID- 14744757 TI - Aurora-A kinase regulates telomerase activity through c-Myc in human ovarian and breast epithelial cells. AB - Aurora-A kinase is frequently overexpressed/activated in human ovarian and breast cancers. A rat mammary tumor model study indicates that alterations of Aurora-A are early events during mammary tumor development (T. M. Goepfert et al., Cancer Res., 62: 4115-4122, 2002), suggesting that Aurora-A plays a pivotal role in transformation. However, the molecular mechanism by which Aurora-A induces ovarian and breast cell transformation remains elusive. Here we show that ectopic expression of Aurora-A induces telomerase activity in human ovarian and breast epithelial cell lines HIOSE118 and MCF-10A. The mRNA and promoter activities of human telomerase reverse transcriptase (hTERT) are stimulated by Aurora-A. Furthermore, we have demonstrated that the c-Myc binding sites of hTERT promoter are required for Aurora-A-induced hTERT promoter activity. Ectopic expression of Aurora-A up-regulates c-Myc. Knockdown of c-Myc by RNA interference attenuates Aurora-A-stimulated hTERT expression and telomerase activity. To our knowledge, these findings demonstrate, for the first time, that Aurora-A induces telomerase activity and hTERT by up-regulation of c-Myc and provides an additional mechanism for the role of Aurora-A in malignant transformation in addition to its cell cycle control. PMID- 14744758 TI - Human papillomavirus type 16 and TP53 mutation in oral cancer: matched analysis of the IARC multicenter study. AB - TP53 mutations were analyzed in 35 human papillomavirus (HPV) type 16 DNA positive cancers of the oral cavity and oropharynx and in 35 HPV DNA-negative cancers matched by subsite, country, sex, age, and tobacco and alcohol consumption. Wild-type TP53 was found more frequently in cancer specimens that contained HPV16 DNA than in those that did not. All 14 HPV16 DNA-positive cancers in HPV16 E6 antibody-positive patients contained wild-type TP53, compared with 50% of corresponding HPV DNA-negative cancers (matched odds ratio, infinity; 95% confidence interval, 1.4- infinity ). In contrast, for HPV16 DNA-positive cancers in E6-negative patients, wild-type TP53 frequency was similar to that in corresponding HPV DNA-negative cancers (matched odds ratio, 1.0; 95% confidence interval, 0.2-5.4). TP53 inactivation is a major mechanism of HPV-related carcinogenesis in the oral cavity and oropharynx. The role of HPV in cancers also containing TP53 mutations remains to be clarified. PMID- 14744760 TI - Kinetics of senescence-associated changes of gene expression in an epithelial, temperature-sensitive SV40 large T antigen model. AB - Replicative senescence limits the number of times primary cells can divide and is therefore regarded as a potential checkpoint for cancer progression. The majority of studies examining changes of gene expression upon senescence have been made with stationary senescent cells. We wanted to study the transition from normal growth to senescence in detail and identify early regulators of senescence by analyzing early changes in global gene expression, using Affymetrix microarrays. For this purpose, we used a murine epithelial senescence model, where senescence is abrogated by SV40 large T antigen and can be induced by using a temperature sensitive form of SV40 large T antigen (SV40ts58). Comparisons were made to wild type SV40 large T antigen-expressing cells and to cells expressing SV40ts58 large T antigen grown to confluence. After removal of genes that are similarly regulated in wild-type and temperature-sensitive SV40 large T antigen-expressing cells, 60% of the remaining genes were shared between cells arrested by inactivation of SV40 T antigen and by confluence. We identified 125 up-regulated and 39 down-regulated candidate genes/expressed sequence tags that are regulated upon SV40 T antigen inactivation and not during heat shock or confluence and classified these based on their kinetic profiles. Our study identified genes that fall into different functional clusters, such as transforming growth factor-beta related genes and transcription factors, and included genes not identified previously as senescence associated. The genes are candidates as early regulators of the senescence checkpoint and may be potential molecular targets for novel anticancer drugs. PMID- 14744759 TI - Involvement of Rel/nuclear factor-kappaB transcription factors in keratinocyte senescence. AB - After a finite doubling number, normal cells become senescent, i.e., nonproliferating and apoptosis resistant. Because Rel/nuclear factor (NF)-kappaB transcription factors regulate both proliferation and apoptosis, we have investigated their involvement in senescence. cRel overexpression in young normal keratinocytes results in premature senescence, as defined by proliferation blockage, apoptosis resistance, enlargement, and appearance of senescence associated beta-galactosidase (SA-beta-Gal) activity. Normal senescent keratinocytes display a greater endogenous Rel/NF-kappaB DNA binding activity than young cells; inhibiting this activity in presenescent cells decreases the number of cells expressing the SA-beta-Gal marker. Normal senescent keratinocytes and cRel-induced premature senescent keratinocytes overexpressed manganese superoxide dismutase (MnSOD), a redox enzyme encoded by a Rel/NF-kappaB target gene. MnSOD transforms the toxic O()(2) into H(2)O(2), whereas catalase and glutathione peroxidase convert H(2)O(2) into H(2)O. Neither catalase nor glutathione peroxidase is up-regulated during cRel-induced premature senescence or during normal senescence, suggesting that H(2)O(2) accumulates. Quenching H(2)O(2) by catalase delays the occurrence of both normal and premature cRel induced senescence. Conversely, adding a nontoxic dose of H(2)O(2) to the culture medium of young normal keratinocytes induces a premature senescence-like state. All these results indicate that Rel/NF-kappaB factors could take part in the occurrence of senescence by generating an oxidative stress via the induction of MnSOD. PMID- 14744761 TI - Role of p12(CDK2-AP1) in transforming growth factor-beta1-mediated growth suppression. AB - p12(CDK2-AP1) (p12) is a growth suppressor isolated from normal keratinocytes. Ectopic expression of p12 in squamous carcinoma cells reversed the malignant phenotype of these cells, in part due an ability of p12 to bind to both DNA polymerase alpha/primase and to cyclin-dependent kinase 2 (CDK2), thereby inhibiting their activities. We report in this article that in normal epithelial cells, transforming growth factor beta1 (TGF-beta1) induces p12 expression transcriptionally, which, in turn, mediates the growth inhibitory activity of TGF beta1. We created inducible p12 antisense HaCaT cell lines [ip12 (-) HaCaT] and showed that selective reduction of cellular p12 resulted in an increase in: (a) CDK2-associated kinase activity; (b) protein retinoblastoma (pRB) phosphorylation; and (c) [(3)H]thymidine incorporation, and partially reversed TGF-beta1-mediated inhibition of CDK2 kinase activity, pRB phosphorylation, and cell proliferation. Furthermore, we generated p12-deficient mouse oral keratinocytes (MOK(p12-/-)) and compared their growth characteristics and response to TGF-beta1 with that of wild-type mouse oral keratinocytes (MOK(WT)). Under normal culture conditions, the number of MOK(p12-/-) in S phase is 2-fold greater than that of MOK(WT). Concomitantly, fewer cells are in G(2) phase in MOK(p12-/-) than that in MOK(WT). Moreover, response to TGF-beta1-mediated growth suppression is compromised in MOK(p12-/-) cells. Mechanistic studies showed that MOK(p12-/-) have increased CDK2 activity and reduced sensitivity to inhibition by TGF-beta1. Collectively our data suggest that p12 plays a role in TGF-beta1 mediated growth suppression by modulating CDK2 activities and pRB phosphorylation. PMID- 14744762 TI - A double-strand break repair defect in ATM-deficient cells contributes to radiosensitivity. AB - The ATM protein, which is mutated in individuals with ataxia telangiectasia (AT), is central to cell cycle checkpoint responses initiated by DNA double-strand breaks (DSBs). ATM's role in DSB repair is currently unclear as is the basis underlying the radiosensitivity of AT cells. We applied immunofluorescence detection of gamma-H2AX nuclear foci and pulsed-field gel electrophoresis to quantify the repair of DSBs after X-ray doses between 0.02 and 80 Gy in confluence-arrested primary human fibroblasts from normal individuals and patients with mutations in ATM and DNA ligase IV, a core component of the nonhomologous end-joining (NHEJ) repair pathway. Cells with hypomorphic mutations in DNA ligase IV exhibit a substantial repair defect up to 24 h after treatment but continue to repair for several days and finally reach a level of unrepaired DSBs similar to that of wild-type cells. Additionally, the repair defect in NHEJ mutants is dose dependent. ATM-deficient cells, in contrast, repair the majority of DSBs with normal kinetics but fail to repair a subset of breaks, irrespective of the initial number of lesions induced. Significantly, after biologically relevant radiation doses and/or long repair times, the repair defect in AT cells is more pronounced than that of NHEJ mutants and correlates with radiosensitivity. NHEJ-defective cells analyzed for survival following delayed plating after irradiation show substantial recovery while AT cells fail to show any recovery. These data argue that the DSB repair defect underlies a significant component of the radiosensitivity of AT cells. PMID- 14744763 TI - Transcriptional regulation of the melanoma prognostic marker melastatin (TRPM1) by MITF in melanocytes and melanoma. AB - Determining the metastatic potential of intermediate thickness lesions remains a major challenge in the management of melanoma. Clinical studies have demonstrated that expression of melastatin/TRPM1 strongly predicts nonmetastatic propensity and correlates with improved outcome, leading to a national cooperative prospective study, which is ongoing currently. Similarly, the melanocytic markers MLANA/MART1 and MITF also have been shown to lose relative expression during melanoma progression. Recent studies have revealed that MITF, an essential transcription factor for melanocyte development, directly regulates expression of MLANA. This prompted examination of whether MITF also might transcriptionally regulate TRPM1 expression. The TRPM1 promoter contains multiple MITF consensus binding elements that were seen by chromatin immunoprecipitation to be occupied by endogenous MITF within melanoma cells. Endogenous TRPM1 expression responded strongly to MITF up- or down-regulation, as did TRPM1 promoter-driven reporters. In addition, MITF and TRPM1 mRNA levels were correlated tightly across a series of human melanoma cell lines. Mice homozygously mutated in MITF showed a dramatic decrease in TRPM1 expression. Finally, the slope of TRPM1 induction by MITF was particularly steep compared with other MITF target genes, suggesting it is a sensitive indicator of MITF expression and correspondingly of melanocytic differentiation. These studies identify MITF as a major transcriptional regulator of TRPM1 and suggest that its prognostic value may be linked to MITF-mediated regulation of cellular differentiation. PMID- 14744764 TI - An Msh2 point mutation uncouples DNA mismatch repair and apoptosis. AB - Mutations in the human DNA mismatch repair gene MSH2 are associated with hereditary nonpolyposis colorectal cancer as well as a significant proportion of sporadic colorectal cancer. The inactivation of MSH2 results in the accumulation of somatic mutations in the genome of tumor cells and resistance to the genotoxic effects of a variety of chemotherapeutic agents. Here we show that the DNA repair and DNA damage-induced apoptosis functions of Msh2 can be uncoupled using mice that carry the G674A missense mutation in the conserved ATPase domain. As a consequence, although Msh2(G674A) homozygous mutant mice are highly tumor prone, the onset of tumorigenesis is delayed as compared with Msh2-null mice. In addition, tumors that carry the mutant allele remain responsive to treatment with a chemotherapeutic agent. Our results indicate that Msh2-mediated apoptosis is an important component of tumor suppression and that certain MSH2 missense mutations can cause mismatch repair deficiency while retaining the signaling functions that confer sensitivity to chemotherapeutic agents. PMID- 14744765 TI - Expression of POT1 is associated with tumor stage and telomere length in gastric carcinoma. AB - Pot1, a telomere end-binding protein in fission yeast and human, is proposed not only to cap telomeres but also to recruit telomerase to the ends of chromosomes. No study has been performed regarding Pot1 expression status in human cancers. Thus, we examined POT1 mRNA expression in 51 gastric cancer (GC) tissues and evaluated telomere length and 3' telomeric overhang signals in 20 of the 51 GC tissues. Quantitative reverse transcription-PCR analysis showed that POT1 expression levels in the tumor relative to those in nonneoplastic mucosa (T/N ratio) were significantly higher in stage III/IV tumors than in stage I/II tumors (P = 0.005). Down-regulation of POT1 (T/n < 0.5) was observed more frequently in stage I/II GC (52.4%, 11 of 21) than in stage III/IV GC (23.3%, 7 of 30; P = 0.033), whereas up-regulation of POT1 (T/n > 2.0) was observed more frequently in stage III/IV GC (33.3%, 10 of 30) than in stage I/II GC (9.5%, 2 of 21; P = 0.048). POT1 expression levels showed decreased in accordance with telomere shortening (r = 0.713, P = 0.002). In-gel hybridization analysis showed that 3' telomeric overhang signals decreased in accordance with decreases in POT1 expression levels (r = 0.696, P = 0.002) and telomere shortening (r = 0.570, P = 0.013). Reduced POT1 expression was observed in GC cell lines with telomeres shortened by treatment with azidothymidine. In addition, inhibition of Pot1 by antisense oligonucleotides led to telomere shortening as well as inhibition of telomerase activity in GC cells. Moreover, inhibition of Pot1 decreased 3' overhang signals and increased the frequency of anaphase bridge (P = 0.0005). These data suggest that Pot1 may play an important role in regulation of telomere length and that inhibition of Pot1 may induce telomere dysfunction. Moreover, changes in POT1 expression levels may be associated with stomach carcinogenesis and GC progression. PMID- 14744766 TI - Moderate hypermutability of a transgenic lacZ reporter gene in Myc-dependent inflammation-induced plasma cell tumors in mice. AB - Mutator phenotypes, a common and largely unexplained attribute of human cancer, might be better understood in mouse tumors containing reporter genes for accurate mutation enumeration and analysis. Previous work on peritoneal plasmacytomas (PCTs) in mice suggested that PCTs have a mutator phenotype caused by Myc deregulating chromosomal translocations and/or phagocyte-induced mutagenesis due to chronic inflammation. To investigate this hypothesis, we generated PCTs that harbored the transgenic shuttle vector, pUR288, with a lacZ reporter gene for the assessment of mutations in vivo. PCTs exhibited a 5.5 times higher mutant frequency in lacZ (40.3 +/- 5.1 x 10(-5)) than in normal B cells (7.36 +/- 0.77 x 10(-5)), demonstrating that the tumors exhibit the phenotype of increased mutability. Studies on lacZ mutant frequency in serially transplanted PCTs and phagocyte-induced lacZ mutations in B cells in vitro indicated that mutant levels in tumors are not determined by exogenous damage inflicted by inflammatory cells. In vitro studies with a newly developed transgenic model of inducible Myc expression (Tet-off/MYC) showed that deregulated Myc sensitizes B cells to chemically induced mutations, but does not cause, on its own, mutations in lacZ. These findings suggested that the hypermutability of PCT is governed mainly by intrinsic features of tumor cells, not by deregulated Myc or chronic inflammation. PMID- 14744767 TI - E6 and E7 oncoproteins induce distinct patterns of chromosomal aneuploidy in skin tumors from transgenic mice. AB - Inactivation of the tumor suppressor genes p53 and Rb are two of the most common genetic alterations in cancer cells. We use a mouse model to dissect the consequences of compromising the function of either of these genes on the maintenance of genomic stability. Thirteen cell lines established from skin tumors of mice expressing either the E6 or E7 oncoprotein of the human papillomavirus (HPV) type 16 under control of the keratin 14 promoter were analyzed by comparative genomic hybridization, spectral karyotyping and fluorescence in situ hybridization, reverse transcription-PCR, and mutation analysis. Deducing from the wealth of molecular cytogenetic data available from human cancers, we hypothesized that the more benign tumors in mice expressing E7 would be distinct from the more aggressive lesions in E6 transgenic mice. Tumorigenesis in E6-expressing mice required specifically the selection and maintenance of cells with extra copies of chromosome 6. Aneuploidy of chromosome 6 was independent of activating mutations in H-ras on chromosome 7. Expression of either E6 or E7 resulted in centrosome aberrations, indicating that each viral oncoprotein interferes independently with the centrosome cycle. Although centrosome aberrations are consistent with development of aneuploidy, no direct correlation was evident between the degree of aneuploidy and the percentage of cells with aberrant centrosomes. Our results show that although aneuploidy and centrosome aberrations are present in tumor cells from mice expressing either E6 or E7, tumorigenesis via E6 requires copy number increases of mouse chromosome 6, which is partially orthologous to human chromosome 3q, a region gained in HPV associated carcinomas. PMID- 14744768 TI - Radiation improves the distribution and uptake of liposomal doxorubicin (caelyx) in human osteosarcoma xenografts. AB - Liposomal drug delivery appears to improve the antitumor effect and reduce toxicity compared with the free drug. The therapeutic index may be improved further by combining cytotoxic drugs and radiotherapy. Successful therapy requires that the cytotoxic agents reach the tumor cells. Therefore, we studied tumor growth and the microdistribution of liposomal doxorubicin (Caelyx) with and without additional ionizing radiation in human osteosarcoma xenografts in athymic mice. Caelyx was injected i.v. 1 day before single or fractionated radiotherapy. Both chemoirradiation regimens induced significant tumor growth delays and worked synergistically. Confocal laser scanning microscopy showed that intact liposomes were located in close proximity to endothelial cells, and the distribution of released doxorubicin was heterogeneous. Before radiotherapy, hardly any doxorubicin was localized in the central parts of the tumor. Radiotherapy increased the tumor uptake of doxorubicin by a factor of two to four, with drug being redistributed farther from the vessels in the tumor periphery and located around vessels in the central parts of the tumor. Colocalization of doxorubicin and hypoxic cells showed no distribution of drug into hypoxic areas. Dynamic contrast-enhanced magnetic resonance imaging (MRI) 1 day before the injection of Caelyx and 2 days after treatment start showed that the combined treatment reduced the vascular volume and the vascular transfer rate of the MRI tracer. The results show that chemoirradiation with Caelyx induces synergistic treatment effects. Improved intratumoral drug uptake and distribution are responsible to some extent for the enhanced antitumor effect. PMID- 14744769 TI - Cyclooxygenase-2 induces EP1- and HER-2/Neu-dependent vascular endothelial growth factor-C up-regulation: a novel mechanism of lymphangiogenesis in lung adenocarcinoma. AB - Cyclooxygenase (COX)-2, the inducible isoform of prostaglandin H synthase, has been implicated in the progression of human lung adenocarcinoma. However, the mechanism underlying COX-2's effect on tumor progression remains largely unknown. Lymphangiogenesis, the formation of new lymphatic vessels, has recently received considerable attention and become a new frontier of tumor metastasis research. Here, we study the interaction between COX-2 and the lymphangiogenic factor, vascular endothelial growth factor (VEGF)-C, in human lung cancer cells and their implication in patient outcomes. We developed an isopropyl-beta-D thiogalactopyranoside-inducible COX-2 gene expression system in human lung adenocarcinoma CL1.0 cells. We found that VEGF-C gene expression but not VEGF-D was significantly elevated in cells overexpressing COX-2. COX-2-mediated VEGF-C up-regulation was commonly observed in a broad array of non-small cell lung cancer cell lines. The use of pharmacological inhibitors or activators and genetic inhibition by EP receptor-antisense oligonucleotides revealed that prostaglandin EP(1) receptor but not other prostaglandin receptors is involved in COX-2-mediated VEGF-C up-regulation. At the mechanistic level, we found that COX 2 expression or prostaglandin E(2) (PGE(2)) treatment could activate the HER 2/Neu tyrosine kinase receptor through the EP(1) receptor-dependent pathway and that this activation was essential for VEGF-C induction. The transactivation of HER-2/Neu by PGE(2) was inhibited by way of blocking the Src kinase signaling using the specific Src family inhibitor, PP1, or transfection with the mutant dominant negative src plasmid. Src kinase was involved in not only the HER-2/Neu transactivation but also the following VEGF-C up-regulation by PGE(2) treatment. In addition, immunohistochemical staining of 59 lung adenocarcinoma specimens showed that COX-2 level was highly correlated with VEGF-C, lymphatic vessels density, and other clinicopathological parameters. Taken together, our results provided evidence that COX-2 up-regulated VEGF-C and promotes lymphangiogenesis in human lung adenocarcinoma via the EP(1)/Src/HER-2/Neu signaling pathway. PMID- 14744770 TI - Coupling tumor necrosis factor-alpha with alphaV integrin ligands improves its antineoplastic activity. AB - Despite the impressive results obtained in animal models, the clinical use of tumor necrosis factor-alpha (TNF) as an anticancer drug is limited by severe toxicity. We have shown previously that targeted delivery of TNF to aminopeptidase N (CD13), a marker of angiogenic vessels, improved the therapeutic index of this cytokine in tumor-bearing mice. To assess whether the vascular targeting approach could be extended to other markers of tumor blood vessels, in this work, we have fused TNF with the ACDCRGDCFCG peptide, a ligand of alpha(V) integrins by recombinant DNA technology. We have found that subnanogram doses of this conjugate are sufficient to induce antitumor effects in tumor-bearing mice when combined with melphalan, a chemotherapeutic drug. Cell adhesion assays and competitive binding experiments with anti-integrin antibodies showed that the Arg Gly-Asp moiety interacts with cell adhesion receptors, including alpha(V)beta(3) integrin, as originally postulated. In addition, ACGDRGDCFCG-mouse TNF conjugate induced cytotoxic effects in standard cytolytic assays, implying that ACGDRGDCFCG mouse TNF conjugate can also bind TNF receptors and trigger death signals. These results indicate that coupling TNF with alpha(V) integrin ligands improves its antineoplastic activity and supports the concept that vascular targeting is a strategy potentially applicable to different endothelial markers, not limited to CD13. PMID- 14744771 TI - Nuclear accumulation of globular actin as a cellular senescence marker. AB - We evaluated the nuclear actin accumulation as a new marker of cellular senescence, using human diploid fibroblast (HDF), chondrocyte primary cultures, Mv1Lu epithelial cells, and Huh7 cancer cells. Nuclear accumulation of globular actin (G-actin) and dephosphorylated cofilin was highly significant in the senescent HDF cells, accompanied with inhibition of LIM kinase (LIMK) -1 activity. When nuclear export of the actin was induced by 12-O tetradecanoylphorbol-13-acetate, DNA synthesis of the senescent cells increased significantly, accompanied with changes of morphologic and biochemical profiles, such as increased RB protein phosphorylation and decreased expressions of p21(WAF1), cytoplasmic p-extracellular signal-regulated kinase 1/2, and caveolins 1 and 2. Significance of these findings was strengthened additionally by the fact that nuclear actin export of young HDF cells was inhibited by the treatment with leptomycin B and mutant cofilin transfection, whose LIMK-1 phosphorylation site was lost, and the old cell phenotypes were duplicated with nuclear actin accumulation, suggesting that nuclear actin accumulation was accompanied with G1 arrest during cellular senescence. The aforementioned changes were observed not only in the replicative senescence but also in the senescence induced by treatment of HDF cells, Mv1Lu, primary culture of human chondrocytes, or Huh7 cells with H-ras virus infection, hydroxyurea, deferoxamine, or H(2)O(2). Nuclear actin accumulation was much more sensitive and an earlier event than the well known, senescence-associated beta-galactosidase activity. PMID- 14744772 TI - The role of p53 in suppression of KSHV cyclin-induced lymphomagenesis. AB - Kaposi's sarcoma-associated herpesvirus (KSHV) encodes a cyclin D homolog, K cyclin, that is thought to promote viral oncogenesis. However, expression of K cyclin in cultured cells not only triggers cell cycle progression but also engages the p53 tumor suppressor pathway, which probably restricts the oncogenic potential of K cyclin. Therefore, to assess the tumorigenic properties of K cyclin in vivo, we transgenically targeted expression of K cyclin to the B and T lymphocyte compartments via the E micro promoter/enhancer. Around 17% of E micro K cyclin animals develop lymphoma by 9 months of age, and all such lymphomas exhibit loss of p53. A critical role of p53 in suppressing K cyclin-induced lymphomagenesis was confirmed by the greatly accelerated onset of B and T lymphomagenesis in all E micro -K cyclin/p53(-/-) mice. However, absence of p53 did not appear to accelerate K cyclin-induced lymphomagenesis by averting apoptosis: E micro -K cyclin/p53(-/-) end-stage lymphomas contained abundant apoptotic cells, and transgenic E micro -K cyclin/p53(-/-) lymphocytes in vitro were not measurably protected from DNA damage-induced apoptosis compared with E micro -K cyclin/p53(wt) cells. Notably, whereas aneuploidy was frequently evident in pre-lymphomatous tissues, end-stage E micro -K cyclin/p53(-/-) tumors showed a near-diploid DNA content with no aberrant centrosome numbers. Nonetheless, such tumor cells did harbor more restricted genomic alterations, such as single-copy chromosome losses or gains or high-level amplifications. Together, our data support a model in which K cyclin-induced genome instability arises early in the pre-tumorigenic lymphocyte population and that loss of p53 licenses subsequent expansion of tumorigenic clones. PMID- 14744773 TI - Endometase/matrilysin-2 in human breast ductal carcinoma in situ and its inhibition by tissue inhibitors of metalloproteinases-2 and -4: a putative role in the initiation of breast cancer invasion. AB - Local disruption of the integrity of both the myoepithelial cell layer and the basement membrane is an indispensable prerequisite for the initiation of invasion and the conversion of human breast ductal carcinoma in situ (DCIS) to infiltrating ductal carcinoma (IDC). We previously reported that human endometase/matrilysin-2/matrix metalloproteinase (MMP) 26-mediated pro-gelatinase B (MMP-9) activation promoted invasion of human prostate carcinoma cells by dissolving basement membrane proteins (Y. G. Zhao et al., J. Biol. Chem., 278: 15056-15064, 2003). Here we report that tissue inhibitor of metalloproteinases (TIMP)-2 and TIMP-4 are potent inhibitors of MMP-26, with apparent K(i) values of 1.6 and 0.62 nM, respectively. TIMP-2 and TIMP-4 also inhibited the activation of pro-MMP-9 by MMP-26 in vitro. The expression levels of MMP-26, MMP-9, TIMP-2, and TIMP-4 proteins in DCIS were significantly higher than those in IDC, atypical intraductal hyperplasia, and normal breast epithelia adjacent to DCIS and IDC by immunohistochemistry and integrated morphometry analysis. Double immunofluorescence labeling and confocal laser scanning microscopy revealed that MMP-26 was colocalized with MMP-9, TIMP-2, and TIMP-4 in DCIS cells. Higher levels of MMP-26 mRNA were also detected in DCIS cells by in situ hybridization. PMID- 14744774 TI - A fragment of histidine-rich glycoprotein is a potent inhibitor of tumor vascularization. AB - In this study, we show that recombinant human histidine-rich glycoprotein (HRGP) has potent antiangiogenic properties as judged from effects on a syngeneic tumor model in C57/bl6 mice. Growth of fibrosarcoma, a very aggressive tumor, was reduced by >60% by HRGP treatment, and tumor angiogenesis was dramatically decreased. Treatment with HRGP led to increased apoptosis and reduced proliferation in the tumors. In contrast, HRGP did not affect apoptosis or DNA synthesis in endothelial cells or tumor cells in vitro. The mechanism of action of HRGP involves rearrangement of focal adhesions and decreased attachment of endothelial cells to vitronectin and, as a consequence, reduced endothelial cell migration. By using truncated versions of HRGP, we demonstrate that the isolated 150 amino acid-residue His/Pro-rich domain, which is also released by spontaneous proteolysis from purified HRGP, mediates the inhibitory effect on chemotaxis. Moreover, the His/Pro-rich domain must be released from HRGP to exert its effect. This study shows for the first time inhibitory effects of HRGP on tumor vascularization in vivo, thus providing proof of concept that HRGP is an angiogenesis inhibitor. PMID- 14744775 TI - herg1 gene and HERG1 protein are overexpressed in colorectal cancers and regulate cell invasion of tumor cells. AB - The acquisition of the capacity to invade surrounding tissues confers a more malignant phenotype to tumor cells and is necessary for the establishment of metastases. The understanding of the molecular mechanisms underlying cell invasion in human solid tumors such as colorectal cancers could provide not only more sensitive prognostic analyses but also novel molecular targets for cancer therapy. We report in this article that K(+) ion channels belonging to the HERG family are important determinants for the acquisition of an invasive phenotype in colorectal cancers. The herg1 gene and HERG1 protein are expressed in many colon cancer cell lines, and the activity of HERG channels modulates colon cancer cell invasiveness. Moreover, the amount of HERG1 protein expressed on the plasma membrane is directly related to the invasive phenotype of colon cancer cells. Finally, both the herg1 gene and HERG1 protein were expressed in a high percentage of primary human colorectal cancers, with the highest incidence occurring in metastatic cancers, whereas no expression could be detected either in normal colonic mucosa or in adenomas. PMID- 14744776 TI - Induction of syndecan-1 expression in stromal fibroblasts promotes proliferation of human breast cancer cells. AB - Infiltrating carcinomas characteristically elicit a reactive stromal response, and accumulating evidence indicates that tumor stroma fibroblasts reciprocally promote tumor development and growth. The cell surface heparan sulfate proteoglycan, syndecan-1 (Sdc1), is thought to function as a coreceptor for growth factor and extracellular matrix interactions, and Sdc1 expression is induced in reactive stromal cells in both mice and man. Mice with a targeted mutation in Sdc1 show reduced tumor development in response to oncogene expression and altered responses to other pathological stimuli that are associated with the induction of stromal Sdc1. Here, we test the hypothesis that Sdc1 is required for the growth-promoting activities of reactive stroma. We found that when highly invasive carcinoma cells (MDA-MB-231) were placed in contact with mouse embryonic fibroblasts (MEFs) in a coculture model, Sdc1 expression was induced. Sdc1 was not induced by less invasive or normal cell lines (T47D and NMuMG). Furthermore, the growth of MDA-MB-231 cells was enhanced by 42% when cocultured with Sdc1+/+ MEFs compared with Sdc1-/- MEFs. When T47D cells were cocultured with fibroblasts that expressed transfected Sdc1, these Sdc1-positive fibroblasts stimulated growth of the breast epithelial cells by 85% compared with untransfected controls. The growth-promoting effect was completely abolished when fibroblasts were transfected with mutant Sdc1 lacking heparan sulfate attachment sites. In conclusion, we have demonstrated that a growth-promoting loop exists between breast cancer cells and their stroma that depends on the activity of glycanated Sdc1. PMID- 14744777 TI - Inhibition of MUC4 expression suppresses pancreatic tumor cell growth and metastasis. AB - The MUC4 mucin is a high molecular weight membrane-bound glycoprotein. It is aberrantly expressed in pancreatic tumors and tumor cell lines with no detectable expression in the normal pancreas. A progressive increase of MUC4 expression has also been observed in pancreatic intraepithelial neoplasia, suggesting its association with disease development. Here, we investigated the consequences of silencing MUC4 expression in an aggressive and highly metastatic pancreatic tumor cell line CD18/HPAF that expresses high levels of MUC4. The expression of MUC4 was down-regulated by the stable integration of a plasmid-construct expressing antisense-MUC4 RNA. A decrease in MUC4 expression, confirmed by Western blot and immunofluorescence analyses, resulted in diminished growth and clonogenic ability of antisense-MUC4-transfected (EIAS19) cells compared with parental, empty vector (ZEO) and sense transfected (ES6) control cells. In addition, EIAS19 cells displayed a significant decrease in tumor growth and metastatic properties when transplanted orthotopically into the immunodeficient mice. In vitro biological assays for motility, adhesion, and aggregation demonstrated a 3-fold decrease in motility of EIAS19 cells compared with control cells, whereas these cells adhered more and showed an increase in cellular aggregation. Interestingly, MUC4 down regulation also correlated with the reduced expression of its putative interacting partner, HER2/neu, in antisense-MUC4-transfected cells. In conclusion, the present work demonstrates, for the first time, a direct association of the MUC4 mucin with the metastatic pancreatic cancer phenotype and provides experimental evidence for a functional role of MUC4 in altered growth and behavioral properties of the tumor cell. PMID- 14744778 TI - Activator protein 2alpha inhibits tumorigenicity and represses vascular endothelial growth factor transcription in prostate cancer cells. AB - Activator protein-2alpha (AP-2) is a transcription factor that regulates proliferation and differentiation in mammalian cells. We have shown previously that although AP-2 is expressed highly in normal prostatic epithelium, its expression is lost in high-grade prostatic intraepithelial neoplasia and prostate cancer, suggesting that loss of AP-2 plays a role in prostate cancer development. We demonstrate that forced AP-2 expression in the prostate cancer cell line LNCaP LN3 (AP-2 negative) inhibited dramatically tumor incidence in nude mice. To identify the genes that might have been responsible for this effect, we used microchip expression array. We found several genes known to be involved in malignancy were deregulated, including the vascular endothelial growth factor (VEGF) gene. Because VEGF was down-regulated by 14.7-fold in the AP-2-transfected cells and because it is a major angiogenic factor in prostate cancer development and progression, we chose to examine the AP-2-VEGF interaction. Our evidence suggests that AP-2 repressed transcriptionally the VEGF promoter by competing with the transcriptional activator Sp3. Loss of AP-2 in prostate cancer cells reduced the AP-2:Sp3 ratio and activated VEGF expression. AP-2 acts as a tumor suppressor gene in prostate cancer. Elucidating the molecular events resulting from loss of AP-2 in the prostate epithelium has implications for the understanding and prevention of the onset of prostate cancer. PMID- 14744779 TI - Novel nanosensors for rapid analysis of telomerase activity. AB - Elevated telomerase levels are found in many malignancies, offering an attractive target for therapeutic intervention and diagnostic or prognostic purposes. Here we describe the use of a novel nanosensor developed for rapid screens of telomerase activity in biological samples. The technique utilizes magnetic nanoparticles that, on annealing with telomerase synthesized TTAGGG repeats, switch their magnet state, a phenomenon readily detectable by magnetic readers. We tested the efficacy of different telomerase inhibitors in crude human and murine samples and show that phosphorylation of telomerase regulates its activity. High-throughput adaptation of the technique by magnetic resonance imaging allowed processing of hundreds of samples within tens of minutes at ultrahigh sensitivities. Together, these studies establish and validate a novel and powerful tool for rapidly sensing telomerase activity and provide the rationale for developing analogous magnetic nanoparticles for in vivo sensing. PMID- 14744780 TI - Capsaicin inhibits in vitro and in vivo angiogenesis. AB - Capsaicin (trans-8-methyl-N-vanillyl-6-nonenamide), a natural product of Capsicum species, is known to induce excitation of nociceptive terminals involved in pain perception. Recent studies have also shown that capsaicin not only has chemopreventive properties against certain carcinogens and mutagens but also exerts anticancer activity. Here, we demonstrated the antiangiogenic activity of capsaicin using in vitro and in vivo assay systems. In vitro, capsaicin inhibited vascular endothelial growth factor (VEGF) -induced proliferation, DNA synthesis, chemotactic motility, and capillary-like tube formation of primary cultured human endothelial cells. Capsaicin inhibited both VEGF-induced vessel sprouting in rat aortic ring assay and VEGF-induced vessel formation in the mouse Matrigel plug assay. Moreover, capsaicin was able to suppress tumor-induced angiogenesis in chick chorioallantoic membrane assay. Capsaicin caused G(1) arrest in endothelial cells. This effect correlated with the down-regulation of the expression of cyclin D1 that led to inhibition of cyclin-dependent kinase 4-mediated phosphorylation of retinoblastoma protein. Signaling experiments show that capsaicin inhibits VEGF-induced p38 mitogen-activated protein kinase, p125(FAK), and AKT activation, but its molecular target is distinct from the VEGF receptor KDR/Flk-1. Taken together, these results demonstrate that capsaicin is a novel inhibitor of angiogenesis and suggest that it may be valuable to develop pharmaceutical drugs for treatment of angiogenesis-dependent human diseases such as tumors. PMID- 14744782 TI - Predictive impact of urokinase-type plasminogen activator: plasminogen activator inhibitor type-1 complex on the efficacy of adjuvant systemic therapy in primary breast cancer. AB - One of the most thoroughly studied systems in relation to its prognostic relevance in patients with breast cancer, is the plasminogen activation system. This system comprises of, among others, the urokinase-type plasminogen activator (uPA) and its main inhibitor (PAI-1). In this study we investigated whether the uPA:PAI-1 complex is associated with the responsiveness of patients with primary breast cancer to adjuvant systemic therapy. Quantitative enzyme-linked immunosorbent assays were used to assess the levels of uPA, PAI-1, and uPA:PAI-1 complex in 1119 tumors of patients with primary invasive breast cancer. These patients were followed for a median follow-up time of 59 months (range, 2-267 months) after the primary diagnosis. Correlations with well-known clinicopathological factors, and univariate and multivariate survival analyses were performed. High uPA:PAI-1 complex levels were correlated with an adverse histological grade, and inversely associated with negative estrogen and progesterone receptor status. High tumor levels of uPA:PAI-1 complex predicted an early relapse in the univariate relapse-free survival analysis (P < 0.001). The multivariate analysis showed that high uPA:PAI-1 complex levels were associated with a decreased relapse-free survival time (P = 0.033), independently of age, tumor size, number of lymph nodes affected, progesterone receptor status, uPA, adjuvant endocrine, and chemotherapy. More important, it was demonstrated that there is a larger benefit from adjuvant chemotherapy for patients with higher versus lower tumor levels of uPA:PAI-1 complex. The results of this study imply that the expression of uPA:PAI-1 complex independently predicts the efficacy of adjuvant chemotherapy in patients with primary breast cancer. PMID- 14744781 TI - Pro-matrix metalloproteinase-2 transfection increases orthotopic primary growth and experimental metastasis of MDA-MB-231 human breast cancer cells in nude mice. AB - The ability to activate pro-matrix metalloproteinase (pro-MMP)-2 via membrane type-MMP is a hallmark of human breast cancer cell lines that show increased invasiveness, suggesting that MMP-2 contributes to human breast cancer progression. To investigate this, we have stably transfected pro-MMP-2 into the human breast cancer cell line MDA-MB-231, which lacks MMP-2 expression but does express its cell surface activator, membrane type 1-MMP. Multiple clones were derived and shown to produce pro-MMP-2 and to activate it in response to concanavalin A. In vitro analysis showed that the pro-MMP-2-transfected clones exhibited an increased invasive potential in Boyden chamber and Matrigel outgrowth assays, compared with the parental cells or those transfected with vector only. When inoculated into the mammary fat pad of nude mice, each of the MMP-2-tranfected clones grew faster than each of the vector controls tested. After intracardiac inoculation into nude mice, pro-MMP-2-transfected clones showed a significant increase in the incidence of metastasis to brain, liver, bone, and kidney compared with the vector control clones but not lung. Increased tumor burden was seen in the primary site and in lung metastases, and a trend toward increased burden was seen in bone, however, no change was seen in brain, liver, or kidney. This data supports a role for MMP-2 in breast cancer progression, both in the growth of primary tumors and in their spread to distant organs. MMP-2 may be a useful target for breast cancer therapy when refinement of MMP inhibitors provides for MMP-specific agents. PMID- 14744783 TI - Matrix metalloproteinase-7 facilitates insulin-like growth factor bioavailability through its proteinase activity on insulin-like growth factor binding protein 3. AB - Matrix metalloproteinase-7 (MMP-7) secreted by cancer cells has been implicated classically in the basement membrane destruction associated with tumor cell invasion and metastasis. Recent epidemiologic studies have established a correlation between high levels of circulating insulin-like growth factor (IGF) and low levels of IGF binding protein 3 (IGFBP-3), and relative risk of developing colon, breast, prostate, and lung cancer, which are known to produce MMP-7. In this study, IGFBP-3 was assessed as a candidate for the physiologic substrate of MMP-7. MMP-7 proteolysis generated four major fragments (26 kDa, 17 kDa, 15.5 kDa, and 15.5 kDa), and two cleavage sites were identified: one at the site of hydrolysis of the K(144)-I(145) peptide bond and one at the R(95)-L(96) peptide bond. The former site is different from the previously reported site of cleavage of IGFBP-3 by other proteases. Addition of IGFBP-3 inhibited IGF-I mediated IGF type 1 receptor (IGF-IR) phosphorylation and activation of the downstream molecule Akt in BALB/c 3T3 fibroblasts overexpressing human IGF-IR (3T3-IGF-IR) and in two human colon cancer cell lines (COLO201 and HT29). Coincubation of the IGF-I/IGFBP-3 complex with MMP-7 restored IGF-I-mediated IGF IR phosphorylation and activation of Akt in these cell lines. The IGF-I signal recovered by MMP-7 protected against apoptosis induced by anoikis in 3T3-IGF-IR cells. These results indicate that MMP-7 proteolysis of IGFBP-3 plays a crucial role in regulating IGF-I bioavailability, thereby promoting cell survival. This mechanism may contribute to the tumorigenesis of MMP-7-producing IGF-IR expressing tumors in the primary site and to organ-specific metastasis in a paracrine manner. PMID- 14744784 TI - Imatinib mesylate resistance through BCR-ABL independence in chronic myelogenous leukemia. AB - Imatinib mesylate (IM) binds to the BCR-ABL protein, inhibiting its kinase activity and effectively controlling diseases driven by this kinase. IM resistance has been associated with kinase mutations or increased BCR-ABL expression. However, disease progression may be mediated by other mechanisms that render tumor cells independent of BCR-ABL. To demonstrate this potential, IM resistant cells were found in chronic myelogenous leukemia patients with continuous BCR-ABL gene expression but undetectable BCR-ABL protein expression. These cells were unresponsive to IM and acquired BCR-ABL-independent signaling characteristics. IM resistance in some patients may be mediated through loss of kinase target dependence. PMID- 14744785 TI - Novel mode of action of tylophorine analogs as antitumor compounds. AB - Tylophorine and its analogs are phenanthroindolizidine alkaloids, several of which have been isolated from the Tylophora genus of plants. Evaluation of (+)-S tylophorine [DCB-3500 (NSC-717335)] and its analog DCB-3503 (NSC-716802) in the National Cancer Institute tumor screen showed a fairly uniform and potent inhibition of cell growth in all 60 cell lines (GI(50) approximately 10(-8) M). To further evaluate the antitumor potential of these compounds, we synthesized four tylophorine analogs, designated DCB-3500, DCB-3501, DCB-3502, and DCB-3503. All four tylophorine analogs exerted potent growth-inhibitory effects against HepG2, a human hepatocellular carcinoma cell line, and KB, a human nasopharyngeal carcinoma cell line. HepG2 cells were more sensitive than KB in terms of loss of clonogenicity. KB variants, which are resistant to etoposide, hydroxyurea, or camptothecin, have similar sensitivities to the tylophorine analogs, as do the parental KB cells. Treatment of nude mice bearing HepG2 tumor xenografts by i.p. injections of DCB-3503 at 6 mg/kg every 8 h on days 0 and 3 resulted in significant tumor growth suppression (P < 0.0001). Unlike conventional antitumor drugs, 3 micro M DCB-3503 did not cause DNA breaks or apoptosis in HepG2 cells. Tylophorine analogs induced albumin expression and decreased alpha-fetoprotein expression in HepG2 cells, which suggests that tylophorine analogs could induce HepG2 differentiation. Tylophorine analogs had an inhibitory effect on cyclic AMP response elements, activator protein-1 sites, or nuclear factor-kappaB binding site-mediated transcriptions. In summary, these tylophorine analogs are a unique class of antitumor compounds that have a mode of action different from known antitumor drugs. PMID- 14744786 TI - Selective growth inhibition of tumor cells by a novel histone deacetylase inhibitor, NVP-LAQ824. AB - We have synthesized a histone deacetylase inhibitor, NVP-LAQ824, a cinnamic hydroxamic acid, that inhibited in vitro enzymatic activities and transcriptionally activated the p21 promoter in reporter gene assays. NVP-LAQ824 selectively inhibited growth of cancer cell lines at submicromolar levels after 48-72 h of exposure, whereas higher concentrations and longer exposure times were required to retard the growth of normal dermal human fibroblasts. Flow cytometry studies revealed that both tumor and normal cells arrested in the G(2)-M phase of the cell cycle after compound treatment. However, an increased sub-G(1) population at 48 h (reminiscent of apoptotic cells) was observed only in the cancer cell line. Annexin V staining data supported our hypothesis that NVP LAQ824 induced apoptosis in tumor and transformed cells but not in normal cells. Western blotting experiments showed an increased histone H3 and H4 acetylation level in NVP-LAQ824-treated cancer cells, suggesting that the likely in vivo target of NVP-LAQ824 was histone deacetylase(s). Finally, NVP-LAQ824 exhibited antitumor effects in a xenograft animal model. Together, our data indicated that the activity of NVP-LAQ824 was consistent with its intended mechanism of action. This novel histone deacetylase inhibitor is currently in clinical trials as an anticancer agent. PMID- 14744787 TI - Resveratrol-induced autophagocytosis in ovarian cancer cells. AB - Resveratrol (3,5,4-trihydroxystilbene), a natural phytoalexin present in grapes, nuts, and red wine, has antineoplastic activities. Several molecular mechanisms have been described to underlie its effects on cells in vitro and in vivo. In the present study, the response of ovarian cancer cells to resveratrol is explored. Resveratrol inhibited growth and induced death in a panel of five human ovarian carcinoma cell lines. The response was associated with mitochondrial release of cytochrome c, formation of the apoptosome complex, and caspase activation. Surprisingly, even with these molecular features of apoptosis, analysis of resveratrol-treated cells by light and electron microscopy revealed morphology and ultrastructural changes indicative of autophagocytic, rather than apoptotic, death. This suggests that resveratrol can induce cell death through two distinct pathways. Consistent with resveratrol's ability to kill cells via nonapoptotic processes, cells transfected to express high levels of the antiapoptotic proteins Bcl-x(L) and Bcl-2 are equally sensitive as control cells to resveratrol. Together, these findings show that resveratrol induces cell death in ovarian cancer cells through a mechanism distinct from apoptosis, therefore suggesting that it may provide leverage to treat ovarian cancer that is chemoresistant on the basis of ineffective apoptosis. PMID- 14744788 TI - Mapping tumor epitope space by direct selection of single-chain Fv antibody libraries on prostate cancer cells. AB - The identification of tumor-specific cell surface antigens is a critical step toward the development of targeted therapeutics for cancer. The epitope space at the tumor cell surface is highly complex, composed of proteins, carbohydrates, and other membrane-associated determinants including post-translational modification products, which are difficult to probe by approaches based on gene expression. This epitope space can be efficiently mapped by complementary monoclonal antibodies. By selecting human antibody gene diversity libraries directly on the surface of prostate cancer cells, we have taken a functional approach to identifying fully human, tumor-specific monoclonal antibodies without prior knowledge of their target antigens. Selection conditions have been optimized to favor tumor-specific antibody binding and internalization. To date, we have discovered >90 monoclonal antibodies that specifically bind and enter prostate cancer cells, with little or no binding to control cells. These antibodies are able to efficiently deliver intracellular payloads when attached to nanoparticles such as liposomes. In addition, a subset of the antibodies displayed intrinsic antiproliferative activity. These tumor-specific internalizing antibodies are likely to be useful for targeted therapeutics either alone or in combination with effector molecules. The antigens they bind constitute a tumor-specific internalizing epitope space that is likely to play a significant role in cancer cell homeostasis. Targeting components of this epitope space may facilitate development of immunotherapeutic and small molecule-based strategies as well as the use of other therapeutic agents that rely upon delivery to the interior of the tumor cell. PMID- 14744789 TI - Determination and modeling of kinetics of cancer cell killing by doxorubicin and doxorubicin encapsulated in targeted liposomes. AB - Various mathematical approaches have been devised to relate the cytotoxic effect of drugs in cell culture to the drug concentration added to the cell culture medium. Such approaches can satisfactorily account for drug response when the drugs are free in solution, but the approach becomes problematic when the drug is delivered in a drug delivery system, such as a liposome. To address this problem, we have developed a simple model that assumes that the cytotoxic potency of a drug is a function of the intracellular drug level in a critical compartment. Upon exposure to drug, cell death commences after a lag time, and the cell kill rate is dependent on the amount of drug in the critical intracellular compartment. The computed number of cells in culture, at any time after exposure to the drug, takes into account the cell proliferation rate, the cell kill rate, the average intracellular drug concentration, and a lag time for cell killing. We have applied this model to compare the cytotoxic effect of doxorubicin (DOX), or DOX encapsulated in a liposome that is targeted to CD44 on B16F10 melanoma cells in culture. CD44 is the surface receptor that binds to hyaluronan and is overexpressed on various cancer cells, including B16F10. We have shown previously that the drug encapsulated in hyaluronan-targeted liposomes was more potent than was the free drug. The model required the determination of the cell-associated DOX after the cells were incubated with various concentrations of the free or the encapsulated drug for 3 h, and the quantification of cell number at various times after exposure to the drug. The uptake of encapsulated drug was greater than that of the free drug, and the ratio of cell association of encapsulated:free drug was 1.3 at 0.5 micro g/ml and increased to 3.3 at 20 micro g/ml DOX. The results demonstrate that the enhanced potency of the encapsulated drug could stem from its enhanced uptake. However, in certain cases, where larger amounts of the free drug were added, such that the intracellular amounts of drug exceeded those obtained from the encapsulated drug, the numbers of viable cells were still significantly smaller for the encapsulated drug. This finding demonstrates that for given amounts of intracellular DOX, the encapsulated form was more efficient in killing B16F10 cells than the free drug. The outcome was expressed in the kinetic model as a 5-6-fold larger rate constant of cell killing potency for the encapsulated drug versus the free drug. The model provides a quantitative framework for comparing the cytotoxic effect in cultured cells when applying the drug in the free form or in a delivery system. PMID- 14744790 TI - Prion protein prevents human breast carcinoma cell line from tumor necrosis factor alpha-induced cell death. AB - To define genetic determinants of tumor cell resistance to the cytotoxic action of tumor necrosis factor alpha (TNF), we have applied cDNA microarrays to a human breast carcinoma TNF-sensitive MCF7 cell line and its established TNF-resistant clone. Of a total of 5760 samples of cDNA examined, 3.6% were found to be differentially expressed in TNF-resistant 1001 cells as compared with TNF sensitive MCF7 cells. On the basis of available literature data, the striking finding is the association of some differentially expressed genes involved in the phosphatidylinositol-3-kinase/Akt signaling pathway. More notably, we found that the PRNP gene coding for the cellular prion protein (PrP(c)), was 17-fold overexpressed in the 1001 cell line as compared with the MCF7 cell line. This differential expression was confirmed at the cell surface by immunostaining that indicated that PrP(c) is overexpressed at both mRNA and protein levels in the TNF resistant derivative. Using recombinant adenoviruses expressing the human PrP(c,) our data demonstrate that PrP(c) overexpression converted TNF-sensitive MCF7 cells into TNF-resistant cells, at least in part, by a mechanism involving alteration of cytochrome c release from mitochondria and nuclear condensation. PMID- 14744791 TI - The role of cytidine deaminase and GATA1 mutations in the increased cytosine arabinoside sensitivity of Down syndrome myeloblasts and leukemia cell lines. AB - Myeloblasts from Down syndrome (DS) children with acute myeloid leukemia (AML) are significantly more sensitive in vitro to 1-beta-D-arabinofuranosylcytosine (ara-C) and generate higher 1-beta-D-arabinofuranosylcytosine 5'-triphosphate (ara-CTP) than non-DS AML myeloblasts. Semiquantitative reverse transcription-PCR analyses demonstrated that transcripts for cytidine deaminase (CDA) were 2.7-fold lower in DS than for non-DS myeloblasts. In contrast, transcripts of cystathionine-beta-synthase and deoxycytidine kinase were a median 12.5- and 2.6 fold higher in DS compared with non-DS myeloblasts. The ratio of deoxycytidine kinase/CDA transcripts significantly correlated with ara-C sensitivities and ara CTP generation. In clinically relevant AML cell line models, high cystathionine beta-synthase transcripts in DS CMK cells were accompanied by 10-fold greater ara C sensitivity and 2.4-fold higher levels of ara-CTP compared with non-DS CMS cells. Overexpression of CDA in non-DS THP-1 cells was associated with a 100-fold decreased ara-C sensitivity and 40-fold decreased ara-CTP generation. THP-1 cells secreted CDA into the incubation media and converted extracellular ara-C completely to 1-beta-D-arabinofuranosyluracil within 30 min. Rapid amplification of 5'-cDNA ends (5'-RACE) and reverse transcription-PCR assays identified short- (sf) and long-form (lf) CDA transcripts in THP-1 cells with different 5' untranslated regions and translational start sites; however, only the latter resulted in the active CDA. Although 5' flanking sequences for both CDA transcripts exhibited promoter activity in reporter gene assays, activity for the CDAlf was low. The presence of several GATA1 binding sites in the CDAsf promoter and the uniform detection of GATA1 mutations in DS megakaryocytic leukemia suggested the potential role of GATA1 in regulating CDA transcription and the CDAsf promoter acting as an enhancer. Transfection of GATA1 into Drosophila Mel-2 cells stimulated the CDAlf promoter in a dose-dependent fashion. Additional identification of the mechanisms of differential expression of genes encoding enzymes involved in ara-C metabolism between DS and non-DS myeloblasts may lead to improvements in AML therapy. PMID- 14744792 TI - Selective potentiation of the hypoxic cytotoxicity of tirapazamine by its 1-N oxide metabolite SR 4317. AB - Tirapazamine (TPZ), a bioreductive drug with selective toxicity for hypoxic cells in tumors, is currently in Phase III clinical trials. It has been suggested to have a dual mechanism of action, both generating DNA radicals and oxidizing these radicals to form DNA breaks; whether the second (radical oxidation) step is rate limiting in cells is not known. In this study we exploit the DNA radical oxidizing ability of the 1-N-oxide metabolite of TPZ, SR 4317, to address this question. SR 4317 at high, but nontoxic, concentrations potentiated the hypoxic (but not aerobic) cytotoxicity of TPZ in all four of the human tumor cell lines tested (HT29, SiHa, FaDu, and A549), thus providing a 2-3-fold increase in the hypoxic cytotoxicity ratio. In potentiating TPZ, SR 4317 was 20-fold more potent than the hypoxic cell radiosensitizers misonidazole and metronidazole but was less potent than misonidazole as a radiosensitizer, suggesting that the initial DNA radicals from TPZ and radiation are different. SR 4317 had favorable pharmacokinetic properties in CD-1 nude mice; coadministration with TPZ provided a large increase in the SR 4317 plasma concentrations relative to that for endogenous SR 4317 from TPZ. It also showed excellent extravascular transport properties in oxic and anoxic HT29 multicellular layers (diffusion coefficient 3 x 10(-6) cm(2)s(-1), with no metabolic consumption). Coadministration of SR 4317 (1 mmol/kg) with TPZ at a subtherapeutic dose (0.133 mmol/kg) significantly enhanced hypoxic cell killing in HT29 tumor xenografts without causing oxic cell killing, and the combination at its maximum tolerated dose was less toxic to hypoxic cells in the retina than was TPZ alone at its maximum tolerated dose. This study demonstrates that benzotriazine mono-N-oxides have potential use for improving the therapeutic utility of TPZ as a hypoxic cytotoxin in cancer treatment. PMID- 14744793 TI - Transcriptional activation of p21(waf1/cip1) by alkylphospholipids: role of the mitogen-activated protein kinase pathway in the transactivation of the human p21(waf1/cip1) promoter by Sp1. AB - Alkylphospholipids (ALKs) are a novel class of antitumor agents with an unknown mechanism of action. The first ALK tested in the clinic, miltefosine, has been approved recently in Europe for the local treatment of patients with cutaneous metastasis. Perifosine, the only available oral ALK, is being studied currently in human cancer clinical trials. We have shown previously that perifosine induces p21(waf1/cip1) in a p53-independent fashion and that induction of p21(waf1/cip1) is required for the perifosine-induced cell cycle arrest because cell lines lacking p21(waf1/cip1) are refractory to perifosine. In this report, we investigated the mechanism by which perifosine induces p21(waf1/cip1) protein expression. We observed that perifosine induces the accumulation of p21(waf1/cip1) mRNA without affecting p21(waf1/cip1) mRNA stability. Using several p21(waf1/cip1) promoter-driven luciferase reporter plasmids, we observed that perifosine activates the 2.4-kb full-length p21(waf1/cip1) promoter as well as a p21 promoter construct lacking p53-binding sites, suggesting that perifosine activates the p21(waf1/cip1) promoter independent of p53. The minimal p21 promoter region required for perifosine-induced p21 promoter activation contains four consensus Sp1-binding sites. Mutations in each particular Sp1 site block perifosine-induced p21(waf1/cip1) expression. Moreover, we showed that perifosine activates the mitogen-activated protein/extracellular signal-regulated kinase pathway, and this activation promotes the phosphorylation of Sp1 in known mitogen activated protein kinase residues (threonine 453 and 739), thereby leading to increased Sp1 binding and enhanced p21(waf1/cip1) transcription. These results represent a novel mechanism by which alkylphospholipids modulate transcription, and may contribute to the discovery of new signal transduction pathways crucial for normal and neoplastic cell cycle control. PMID- 14744794 TI - A novel orally bioavailable inhibitor of kinase insert domain-containing receptor induces antiangiogenic effects and prevents tumor growth in vivo. AB - A strategy for antagonizing vascular endothelial growth factor (VEGF) -induced angiogenesis is to inhibit the kinase activity of its receptor, kinase insert domain-containing receptor (KDR), the first committed and perhaps the last unique step in the VEGF signaling cascade. We synthesized a novel ATP-competitive KDR tyrosine kinase inhibitor that potently suppresses human and mouse KDR activity in enzyme (IC(50) = 7.8-19.5 nM) and cell-based assays (IC(50) = 8 nM). The compound was bioavailable in vivo, leading to a dose-dependent decrease in basal- and VEGF-stimulated KDR tyrosine phosphorylation in lungs from naive and tumor bearing mice (IC(50) = 23 nM). Pharmacokinetics and pharmacodynamics guided drug dose selection for antitumor efficacy studies. HT1080 nude mice xenografts were treated orally twice daily with vehicle, or 33 or 133 mg/kg of compound. These doses afforded trough plasma concentrations approximately equal to the IC(50) for inhibition of KDR autophosphorylation in vivo for the 33 mg/kg group, and higher than the IC(99) for the 133 mg/kg group. Chronic treatment at these doses was well-tolerated and resulted in dose-dependent inhibition of tumor growth, decreased tumor vascularization, decreased proliferation, and enhanced cell death. Antitumor efficacy correlated with inhibition of KDR tyrosine phosphorylation in the tumor, as well as in a surrogate tissue (lung). Pharmacokinetics and pharmacodynamics assessment indicated that the degree of tumor growth inhibition correlated directly with the extent of inhibition of KDR tyrosine phosphorylation in tumor or lung at trough. These observations highlight the need to design antiangiogenic drug regimens to ensure constant target suppression and to take advantage of PD end points to guide dose selection. PMID- 14744795 TI - Adjuvant-mediated tumor regression and tumor-specific cytotoxic response are impaired in MyD88-deficient mice. AB - The Mycobacterium bovis bacillus Calmette-Guerin cell-wall skeleton (BCG-CWS) activates Toll-like receptor (TLR) 2 and TLR4, but unlike the typical TLR4 agonist bacterial lipopolysaccharide barely induces type 1 IFN. BCG-CWS has been used for adjuvant immunotherapy for patients with cancer. We investigated the adjuvant potential of BCG-CWS for induction of CTLs subsequent to TLR-mediated dendritic cell (DC) maturation, using a syngeneic mouse tumor model (B16 melanoma in C57BL/6). We evaluated the retardation of tumor growth and cytotoxic response in wild-type and MyD88-/- mice immunized with tumor debris and/or BCG-CWS. Delays in tumor growth and cytotoxic response were induced by immunization with a mixture of BCG-CWS emulsion and the tumor. BCG-CWS was capable of activating DCs ex vivo by the criteria of CD80/CD86 up-regulation and cytokine (interleukin-12, tumor necrosis factor-alpha) induction. Efficient tumor suppression and ex vivo cytokine induction did not occur in MyD88-deficient mice and cells, suggesting that the MyD88 adapter is crucial for induction of tumor cytotoxicity. Because TLR4 is involved in both MyD88-dependent and -independent pathways and the latter affects DC maturation, our findings indicate that both pathways cooperate to induce CTL-based tumor immunity. PMID- 14744796 TI - The adrenal androgen androstenediol is present in prostate cancer tissue after androgen deprivation therapy and activates mutated androgen receptor. AB - Despite an initial response to androgen deprivation therapy, prostate cancer (PCa) progresses eventually from an androgen-dependent to an androgen-independent phenotype. One of the mechanisms of relapse is antiandrogen withdrawal phenomenon caused by mutation of 877th amino acid of androgen receptor (AR). In the present study, we established a method to measure the concentration of androstenediol (adiol) in prostate tissue. We found that adiol maintains a high concentration in PCa tissue even after androgen deprivation therapy. Furthermore, adiol is a stronger activator of mutant AR in LNCaP PCa cells and induces more cell proliferation, prostate-specific antigen (PSA) mRNA expression, and PSA promoter than dihydrotestosterone (DHT). Because antiandrogen, bicalutamide, blocked adiol activity in LNCaP cells, it was suggested that adiol effect was mediated through AR. However, high concentration of bicalutamide was necessary to block completely adiol activity. These effects were specific to LNCaP cells because adiol had less effect in PC-3 PCa cells transfected with wild-type AR than DHT and had similar effect in PC-3 cells transfected with mutant AR. The mechanism that adiol activates mutant AR in LNCaP cells did not result from the increased affinity to mutant AR or from AR's association with coactivator ARA70. However, low concentration of adiol induced more AR nuclear translocation than DHT in LNCaP cells and not PC-3 cells transfected with AR. These results indicate that adiol may cause the progression of PCa even after hormone therapy. PMID- 14744797 TI - Do human myeloma cells directly produce the receptor activator of nuclear factor kappaB ligand (RANKL) or induce RANKL in the bone marrow microenvironment? PMID- 14744798 TI - Identification and clinical relevance of receptor activator of nuclear factor kappaB ligand expression of myeloma cells. PMID- 14744799 TI - Correspondence re: T. Zhang et al., Evidence that APC regulates survivin expression: a possible mechanism contributing to the stem cell origin of colon cancer. Cancer Res., 61: 8664-8667, 2001. PMID- 14744800 TI - Regulation of microglial inflammatory response by sodium butyrate and short-chain fatty acids. AB - 1. Recent studies have shown that sodium butyrate and other short-chain fatty acids (SCFAs) can prevent inflammation in colon diseases. Our aim was to elucidate whether sodium butyrate and SCFAs regulate the inflammatory responses in different neural inflammation models in cell cultures. 2. Inflammatory responses to LPS-induced microglial activation were recorded by the secretion of nitric oxide (NO) and cytokines IL-6 and TNF-alpha and related to the changes in the DNA-binding activities of NF-kappaB complex. 3. We observed that sodium butyrate is strongly anti-inflammatory against LPS-induced responses in rat primary microglia as well as in hippocampal slice cultures and in neural cocultures of microglial cells, astrocytes and cerebellar granule neurons. 4. In murine N9 microglial cell line, instead, sodium butyrate and other SCFAs (propionate, valerate and caproate) enhanced the LPS-induced inflammatory response. 5. The pretreatment with butyrate before LPS exposure induced an equal or more enhanced response than simultaneous exposure with butyrate and LPS. This indicates that butyrate induces an adaptative response against microglial activation. 6. We also observed that butyrate treatment both in transformed N9 cells and in hippocampal slice cultures downregulates the NF-kappaB-binding capacity induced by LPS stimulation. 7. Our results show that butyrate is anti inflammatory in primary, brain-derived microglial cells, as observed recently in colon diseases, but proinflammatory in transformed, proliferating N9 microglial cells, which may be related to the anticancer properties of butyrate observed in tumor cells. PMID- 14744801 TI - The endocannabinoid system: a general view and latest additions. AB - After the discovery, in the early 1990s, of specific G-protein-coupled receptors for marijuana's psychoactive principle Delta(9)-tetrahydrocannabinol, the cannabinoid receptors, and of their endogenous agonists, the endocannabinoids, a decade of investigations has greatly enlarged our understanding of this altogether new signalling system. Yet, while the finding of the endocannabinoids resulted in a new effort to reveal the mechanisms regulating their levels in the brain and peripheral organs under physiological and pathological conditions, more endogenous substances with a similar action, and more molecular targets for the previously discovered endogenous ligands, anandamide and 2-arachidonoylglycerol, or for some of their metabolites, were being proposed. As the scenario becomes subsequently more complicated, and the experimental tasks to be accomplished correspondingly more numerous, we briefly review in this article the latest 'additions' to the endocannabinoid system together with earlier breakthroughs that have contributed to our present knowledge of the biochemistry and pharmacology of the endocannabinoids. PMID- 14744802 TI - Noradrenaline inhibits pacemaker currents through stimulation of beta 1 adrenoceptors in cultured interstitial cells of Cajal from murine small intestine. AB - 1. Interstitial cells of Cajal (ICCs) are pacemaker cells that activate the periodic spontaneous inward currents (pacemaker currents) responsible for the production of slow waves in gastrointestinal smooth muscle. The effects of noradrenaline on the pacemaker currents in cultured ICCs from murine small intestine were investigated by using whole-cell patch-clamp techniques at 30 degrees C. 2. Under current clamping, ICCs had a mean resting membrane potential of -58+/-5 mV and produced electrical slow waves. Under voltage clamping, ICCs produced pacemaker currents with a mean amplitude of -410+/-57 pA and a mean frequency of 16+/-2 cycles min(-1). 3. Under voltage clamping, noradrenaline inhibited the amplitude and frequency of pacemaker currents and increased resting currents in the outward direction in a dose-dependent manner. These effects were reduced by intracellular GDP beta S. 4. Noradrenaline-induced effects were blocked by propranolol (beta-adrenoceptor antagonist). However, neither prazosin (alpha(1)-adrenoceptor antagonist) nor yohimbine (alpha(2)-adrenoceptor antagonist) blocked the noradrenaline-induced effects. Phenylephrine (alpha(1) adrenoceptor agonist) had no effect on the pacemaker currents, whereas isoprenaline (beta-adrenoceptor agonist) mimicked the effect of noradrenaline. Atenolol (beta(1)-adrenoceptor antagonist) blocked the noradrenaline-induced effects, but butoxamine (beta(2)-adrenoceptor antagonist) did not. In addition, BRL37344 (beta(3)-adrenoceptor agonist) had no effect on pacemaker currents. 5. 9 (Tetrahydro-2-furanyl)-9H-purine-6-amine (SQ-22536; adenylate cyclase inhibitor) and a myristoylated protein kinase A inhibitor did not inhibit the noradrenaline induced effects and 8-bromo-cAMP had no effects on pacemaker currents. 8-Bromo cGMP and SNAP inhibited pacemaker currents and these effects of SNAP were blocked by 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ; a guanylate cyclase inhibitor). However, ODQ did not block the noradrenaline-induced effects. 6. Neither tetraethylammonium (a voltage-dependent K(+) channel blocker), apamin (a Ca(2+)-dependent K(+) channel blocker) nor glibenclamide (an ATP-sensitive K(+) channel blocker) blocked the noradrenaline-induced effects. 7. The results suggest that noradrenaline-induced stimulation of beta(1)-adrenoceptors in the ICCs inhibits pacemaker currents, and that this is mediated by the activation of G-protein. Neither adenylate cyclase, guanylate cyclase nor a K(+) channel dependent pathway are involved in this effect of noradrenaline. PMID- 14744803 TI - Endothelins contribute towards nociception induced by antigen in ovalbumin sensitised mice. AB - 1. The contribution of endogenous endothelins to nociceptive responses elicited by ovalbumin (OVA) in the hind-paw of mice sensitised to this antigen (50 microg OVA+5 mg Al(OH)(3), s.c., 14 days beforehand) was investigated. 2. Sensitised mice exhibited greater nocifensive responsiveness to intraplantar (i.pl.) OVA (total licking time over first 30 min: 85.2+/-14.6 s at 0.3 microg; 152.6+/-35.6 s at 1 microg) than nonsensitised animals (29.3+/-7.4 s at 1 microg). Nocifensive responses of sensitised mice to 0.3 microg OVA were inhibited by morphine (3 mg kg(-1), s.c.) or local depletion of mast cells (four daily i.pl. injections of compound 48/80). 3. Pretreatment with i.v. bosentan (mixed ET(A)/ET(B) receptor antagonist; 52 micromol kg(-1)) or A-122722.5 (selective ET(A) receptor antagonist; 6 micromol kg(-1)) reduced OVA-induced licking from 124.8+/-20.6 s to 45.7+/-13.0 s and 64.2+/-12.1 s, respectively, whereas A-192621.1 (selective ET(B) receptor antagonist; 25 micromol kg(-1)) enhanced them to 259.2+/-39.6 s. 4. Local i.pl. pretreatment with BQ-123 or BQ-788 (selective ET(A) or ET(B) receptor antagonists, respectively, each at 3 nmol) reduced OVA-induced licking (from 106.2+/-15.2 to 57.0+/-9.4 s and from 118.6+/-10.5 to 76.8+/-14.7 s, respectively). Sarafotoxin S6c (selective ETB receptor agonist, 30 pmol, i.pl., 30 min after OVA) induced nocifensive responses in OVA-sensitised, but not in nonsensitised, animals. 5. Compound 48/80 (0.3 microg, i.pl.) induced nocifensive responses per se and potentiated those induced by i.pl. capsaicin (0.1 microg). Treatment with BQ-123 (3 nmol, i.pl.) reduced only the hyperalgesic effect of compound 48/80, whereas BQ-788 (3 nmol) was ineffective. 6. Thus, immune-mediated Type I hypersensitivity reactions elicit mast cell- and endothelin-dependent nociception in the mouse hind-paw, which are mediated locally by both ET(A) and ET(B) receptors. The nocifensive response to antigen is amenable to blockade by systemic treatment with dual ET(A)/ET(B) or selective ET(A) receptor antagonists, but is sharply potentiated by systemic selective ET(B) receptor antagonist treatment. The apparently distinct roles played by ET(B) receptors in this phenomenon at local and other sites remain to be characterised. PMID- 14744804 TI - Neuroprotective effects of M826, a reversible caspase-3 inhibitor, in the rat malonate model of Huntington's disease. AB - 1. Caspases, key enzymes in the apoptosis pathway, have been detected in the brain of HD patients and in animal models of the disease. In the present study, we investigated the neuroprotective properties of a new, reversible, caspase-3 specific inhibitor, M826 (3-([(2S)-2-[5-tert-butyl-3-[[(4-methyl-1,2,5-oxadiazol 3-yl)methyl]amino]-2-oxopyrazin-1(2H)-yl]butanoyl]amino)-5-[hexyl(methyl)amino]-4 oxopentanoic acid), in a rat malonate model of HD. 2. Pharmacokinetic and autoradiography studies after intrastriatal (i.str.) injection of 1.5 nmol of M826 or its tritiated analogue [(3)H]M826 indicated that the compound diffused within the entire striatum. The elimination half-life (T(1/2)) of M826 in the rat striatum was 3 h. 3. I.str. injection of 1.5 nmol of M826 10 min after malonate infusion induced a significant reduction (66%) in the number of neurones expressing active caspase-3 in the ipsilateral striatum. 4. Inhibition of active caspase-3 translated into a significant but moderate reduction (39%) of the lesion volume, and of cell death (24%), 24 h after injury. The efficacy of M826 at inhibiting cell death was comparable to that of the noncompetitive NMDA receptor antagonist MK801. 5. These data provide in vivo proof-of-concept of the neuroprotective effects of reversible caspase-3 inhibitors in a model of malonate induced striatal injury in the adult rat. PMID- 14744806 TI - Donepezil modulates nicotinic receptors of substantia nigra dopaminergic neurones. AB - 1. The effects of donepezil, one of the most common cholinesterase inhibitors used for treatment of Alzheimer's disease, were studied on nicotinic receptors (nAChRs)-mediated postsynaptic currents, in dopaminergic neurons of the substantia nigra pars compacta, using the patch-clamp recording technique in slice preparations. 2. Donepezil (10-100 microM) selectively and reversibly depressed nicotine currents, induced by brief puffer pulses, through a glass micropipette positioned above the slice. 3. The peak amplitude fading of the responses generated by repeated test applications of low doses of nicotine was accelerated by donepezil, while it slowed the recovery of nicotine currents after a large, desensitising, dose of the same agonist. 4. Donepezil depressed even maximal responses to nicotine, revealing a noncompetitive mechanism of action; moreover, the inhibition of nAChRs was voltage and time independent. 5. Pretreatment with vesamicol or methamidophos did not prevent the reduction of nicotine-induced currents. The data indicated direct effect on nAChR, independent from the activity of donepezil as cholinesterase inhibitor. PMID- 14744805 TI - Role of cyclic GMP on inhibition by nitric oxide donors of human eosinophil chemotaxis in vitro. AB - 1. This study was designed to investigate the effects of the nitric oxide (NO) donors sodium nitroprusside (SNP), 3-morpholinosydnonimine (SIN-1) and S-nitroso N-acetylpenicillamine (SNAP) on N-formyl-L-methionyl-L-leucyl-phenylalanine (fMLP, 1 x 10(-7) M)-induced human eosinophil chemotaxis, cyclic guanosine-3',5' monophosphate (cGMP) levels, protein nitration and cytotoxicity. 2. Human eosinophils were exposed to SNP, SIN-1 and SNAP (0.001-1.0 mM) for either short (10 min) or prolonged (90 min) time periods. Exposition of eosinophils with these NO donors significantly inhibited the eosinophil chemotaxis irrespective of whether cells were exposed to these agents for 10 or 90 min. No marked differences were detected among them regarding the profile of chemotaxis inhibition. 3. Exposition of eosinophils to SNP, SIN-1 and SNAP (0.001-1.0 mM) markedly elevated the cGMP levels above basal levels, but the 90-min exposition resulted in significantly higher levels compared with the 10-min protocols (5.3+/ 0.6 and 2.6+/-0.2 nM 1.5 x 10(6) cells(-1), respectively). The cGMP levels achieved with SNAP were greater than SNP and SIN-1. 4. The NO donors did not induce cell toxicity in any experimental condition used. Additionally, eosinophils exposed to SNP, SIN-1 and SNAP (1.0 mM each) either for 10 or 90 min did not show any tyrosine nitration in conditions where a strong nitration of bovine serum albumin was observed. 5. Our findings show that inhibitory effects of fMLP-induced human eosinophil chemotaxis by NO donors at short or prolonged exposition time were accompanied by significant elevations of cGMP levels. However, additional elevations of cGMP levels do not change the functional profile (chemotaxis inhibition) of stimulated eosinophils. PMID- 14744807 TI - Regional haemodynamic effects of cyclosporine A, tacrolimus and sirolimus in conscious rats. AB - 1. The observation that the immunosuppressants, cyclosporine A (CsA) and tacrolimus, have pressor effects, but sirolimus does not, has led to an hypothesis that generalised sympathoexcitation, resulting from inhibition of calcineurin by CsA and tacrolimus underlies their pressor effects, because sirolimus does not inhibit calcineurin. It is unknown if sirolimus has haemodynamic actions not accompanied by a pressor effect, and whether or not the pressor effects of CsA and tacrolimus are accompanied by similar haemodynamic changes. Therefore, the first aim of our studies was to investigate these possibilities in conscious, chronically-instrumented, male, Sprague-Dawley rats. 2. CsA (5.9 mg kg(-1) bolus i.v.) caused rapid-onset, prolonged hypertension, tachycardia and mesenteric vasoconstriction. There was a slower onset renal vasoconstriction, but no significant change in hindquarters vascular conductance; all the effects of CsA were significantly greater than those of vehicle. CsA given by infusion (over 30 min or 2 h) caused changes qualitatively similar to those above. Repeated administration of CsA over 4 days did not enhance its cardiovascular effects. 3. Pretreatment with the angiotensin (AT(1)) receptor antagonist, losartan, and the endothelin (ET(A) and ET(B)) receptor antagonist, SB 209670, reduced the pressor and mesenteric vasoconstrictor effects of CsA. Additional administration of the alpha-adrenoceptor antagonist, phentolamine, completely inhibited the cardiovascular effects of CsA. 4. Tacrolimus (450 microg kg(-1) bolus i.v.) caused similar peak pressor and tachycardic effects to CsA, but these were much slower in onset, and were maximal when there were no significant regional vasoconstrictions, indicating that the pressor effect was probably due to a rise in cardiac output. However, although propranolol reversed the tachycardic effect of tacrolimus, it did not influence the pressor response. 5. Sirolimus (450 microg kg(-1) bolus i.v.) had no tachycardic action, and only a modest, transient pressor effect, accompanied by equally brief reductions in renal, mesenteric, and hindquarters vascular conductances. 6. The differences between the regional haemodynamic profiles of equipressor doses of CsA and tacrolimus, and the finding that sirolimus has significant cardiovascular actions, indicate that generalised sympathoexcitation, resulting from calcineurin inhibition (with CsA and tacrolimus), is unlikely to be the sole explanation of their pressor effects. PMID- 14744808 TI - Sildenafil (Viagra) reduces arrhythmia severity during ischaemia 24 h after oral administration in dogs. AB - Sildenafil (Viagra) prolongs repolarisation in cardiac muscle, an effect that could lead to ventricular fibrillation (VF). Sildenafil (2 mg kg(-1)) was given by mouth to 12 mongrel dogs and, 24 h later, these dogs were anaesthetised, thoracotomised and subjected to a 25 min occlusion of the anterior descending coronary artery. Haemodynamic parameters were similar in this and the control group, but there were fewer and less serious ventricular arrhythmias during occlusion in the sildenafil group (VF 17 vs 60%; ventricular premature beats 140+/-52 vs 437+/-127% and episodes of ventricular tachycardia 4.0+/-3.2 vs 19.3+/-7.7%, all P<0.05). However, reperfusion VF and indices of ischaemia severity (epicardial ST-segment mapping, inhomogeneity) were not modified by the drug. Sildenafil increased the QT interval, especially during ischaemia. Our conclusion is that ischaemia-induced ventricular arrhythmias are reduced by sildenafil, but this protection is less pronounced than that following cardiac pacing or exercise. PMID- 14744809 TI - Histamine-induced inhibition of leukotriene biosynthesis in human neutrophils: involvement of the H2 receptor and cAMP. AB - 1. Histamine is generally regarded as a pro-inflammatory mediator in diseases such as allergy and asthma. A growing number of studies, however, suggest that this autacoid is also involved in the downregulation of human polymorphonuclear leukocyte (PMN) functions and inflammatory responses through activation of the Gs coupled histamine H(2) receptor. 2. We report here that histamine inhibits thapsigargin- and ligand (PAF and fMLP)-induced leukotriene (LT) biosynthesis in human PMN in a dose-dependent manner. 3. The suppressive effect of histamine on LT biosynthesis was abrogated by the histamine H(2) receptor antagonists cimetidine, ranitidine, and tiotidine. In contrast, the histamine H(1), H(3), and H(4) receptor antagonists used in this study were ineffective in counteracting the inhibitory effect of histamine on the biosynthesis of LT in activated human PMN. 4. The inhibition of LT biosynthesis by histamine was characterized by decreased arachidonic acid release and 5-lipoxygenase translocation to the nuclear membrane. 5. Incubation of PMN with the cAMP-dependent protein kinase (PKA) inhibitor N-[2-(p-bromocinnamylamino)ethyl]-5-isoquinoline-sulfonamide prevented the inhibitory effect of histamine on LT biosynthesis, suggesting an important role for PKA in this effect of histamine on LT biosynthesis in PMN. 6. These data provide the first evidences that, similarly to adenosine and prostaglandin E(2), histamine is a potent suppressor of LT biosynthesis, and support the concept that histamine may play a dual role in the regulation of inflammation. PMID- 14744810 TI - Circadian rhythm of dihydrouracil/uracil ratios in biological fluids: a potential biomarker for dihydropyrimidine dehydrogenase levels. AB - 1. In many cancer patients, 5-fluorouracil (5-FUra) treatment is toxic and even causes death. Nevertheless, all patients are subjected to a standard therapy regimen because there is no reliable way to identify beforehand those patients who are predisposed to 5-FUra-induced toxicity. In this study, we identified the dihydrouracil/uracil (UH2/Ura) ratio in plasma or urine as a potential biomarker reflecting the activity of dihydropyrimidine dehydrogenase (DPD), the rate limiting enzyme in 5-FUra metabolism. 2. UH2/Ura ratios were measured by high performance liquid chromatography tandem triple quadrupole mass spectrometry (HPLC-MS/MS) in both healthy subjects (n=55) and in patients (n=20) diagnosed with grade I/II gestational trophoblastic tumours. In addition, rats (n=18) were used as an animal model to verify a correlation between UH2/Ura ratios and DPD levels in the liver. 3. A significant circadian rhythm was observed in UH2/Ura ratios in healthy subjects, whereas a disrupted rhythm occurred in cancer patients who were continuously infused with a high dose of 5-FUra. In rats, UH2/Ura ratios, liver DPD levels and PBMC DPD levels showed a definite circadian rhythm. Significant linear correlations with liver DPD levels were demonstrated for plasma UH2/Ura ratios (r=0.883, P<0.01), urine UH2/Ura ratios (r=0.832, P<0.01) and PBMC DPD levels (r=0.859, P<0.01). 4. The UH2/Ura ratio in biological fluid was significantly correlated with liver DPD levels; hence, this ratio could be a potential biomarker to identify patients with a deficiency in DPD. PMID- 14744811 TI - Particular sensitivity to calcium channel blockers of the fast inward voltage dependent sodium current involved in the invasive properties of a metastastic breast cancer cell line. AB - 1. A voltage-dependent sodium current has been described in the highly invasive breast cancer cell line MDA-MB-231. Its activity is associated with the invasive properties of the cells. The aim of our study was to test whether this current (I(Na)) is sensitive to three representative calcium channel blockers: verapamil, diltiazem and nifedipine. I(Na) was studied in patch-clamp conditions. 2. I(Na) was sensitive to verapamil (IC(50)=37.6+/-2.5 microM) and diltiazem (53.2+/-3.6 microM), while it was weakly sensitive to nifedipine. 3. The tetrodotoxin (TTX) concentration, which fully blocks I(Na) (30 microM), did not affect cell proliferation. Diltiazem and verapamil, at concentrations that do not fully block I(Na), strongly reduced cell proliferation, suggesting, regarding proliferation, that these molecules act on targets distinct from sodium channels. These targets are probably not other ionic channels, since the current measured at the end of a 500 ms long pulse in the voltage range between -60 and +40 mV was unaffected by verapamil and diltiazem. 4. We conclude that the sodium channel expressed in MDA MB-231 cells is sensitive to several calcium channel blockers. The present study also underlines the danger of concluding to the possible involvement of membrane channel proteins in any phenomenon on the sole basis of pharmacology, and without an electrophysiological confirmation. PMID- 14744812 TI - E-ring 8-isoprostanes inhibit ACh release from parasympathetic nerves innervating guinea-pig trachea through agonism of prostanoid receptors of the EP3-subtype. AB - 1. In the present study, we examined the effect of E-ring 8-isoprostanes on cholinergic neurotransmission in guinea-pig trachea and identified the receptor(s) involved. As isoprostanes are isomeric with prostaglandins, PGE(2) and sulprostone (a selective EP(3)-receptor agonist) were examined in parallel. 2. 8-Iso-PGE(1), 8-iso-PGE(2) (0.1 nm-1 microM), sulprostone (1 nm-1 microM) and PGE(2) (1 microM) suppressed EFS-evoked [(3)H]ACh release from guinea-pig trachea in a concentration-dependent manner, producing 39.5, 53.9, 61.2 and 59.9% inhibition, respectively, at 1 microM. It should be noted that an established maximum effective concentration was not determined. 3. Neither SQ 29,548 (1 microm; a TP-receptor antagonist) nor AH 6809 (10 microM; an EP(1)-/EP(2)-/DP receptor antagonist) reversed the inhibitory effect of these compounds. 4. L 798,106, a novel and highly selective EP(3)-receptor antagonist, produced a parallel shift to the right of the concentration-response curves that described the inhibitory action of sulprostone on EFS-evoked contractile responses in guinea-pig vas deferens (an established EP(3)-receptor-expressing tissue), from which a mean pA(2) of 7.48 was derived. On guinea-pig trachea, L-798,106 also antagonised sulprostone-induced inhibition of EFS-induced twitch responses, with similar potency (mean pA(2)=7.82). 5. The inhibitory effects of 8-iso-PGE(1), 8 iso-PGE(2), sulprostone and PGE(2) on EFS-induced [(3)H]ACh release was blocked by L-798,106 at a concentration (10 microM) that binds only weakly to human recombinant EP(1)-, EP(2)- and EP(4)-receptor subtypes expressed in HEK 293 cells. 6. These data suggest that E-ring 8-isoprostanes, PGE(2) and sulprostone inhibit EFS-evoked [(3)H]ACh release from cholinergic nerves innervating guinea pig trachea, by interacting with prejunctional prostanoid receptors of the EP(3) subtype. PMID- 14744813 TI - Comparison of U46619-, endothelin-1- or phenylephrine-induced changes in cellular Ca2+ profiles and Ca2+ sensitisation of constriction of pressurised rat resistance arteries. AB - 1. In pressurised rat mesenteric small arteries (50 mmHg), we examined the effects of stimulation with U46619, endothelin-1 (ET-1) or phenylephrine (PE) on changes in vessel diameter, global [Ca(2+)](i), individual smooth muscle cell [Ca(2+)](i) and Ca(2+)-sensitisation of contraction. 2. U46619 or ET-1 gave tonic diameter reductions, whereas PE-stimulated vessels gave tonic contractions or initial vasoconstrictions followed by diameter oscillations. Global [Ca(2+)](i) changes were transient for each agonist, with tonic constrictions being accompanied by maintained submaximal global [Ca(2+)](i) levels. 3. U46619, ET-1 or PE tonic constrictions were accompanied by apparently asynchronous [Ca(2+)](i) waves in individual smooth muscle cells of the vessel wall, as examined by confocal fluorescent microscopy. In vessels exhibiting vasomotion to PE, some apparent synchrony of activation of individual cells was evident; however, this was incomplete with many cells responding out of phase with their neighbours. 4. In alpha-toxin-permeabilised preparations, agonist-induced Ca(2+)-sensitisation of constriction at submaximal Ca(2+) (pCa6.7) in the presence of GTP was greater with U46619 or ET than PE. 5. We conclude that, in pressurised mesenteric arteries, (i) a general feature of receptor-coupled constriction is the generation of periodic smooth muscle [Ca(2+)](i) waves; (ii) complete synchrony of Ca(2+) oscillations between smooth muscle cells is not a prerequisite for receptor-coupled vasomotion; (iii) varied Ca(2+)-sensitising actions of agonists may partly determine tonic or phasic vessel responses to different stimuli. PMID- 14744814 TI - Potential mechanisms for the enhancement of HERG K+ channel function by phospholipid metabolites. AB - 1. Phospholipid metabolites lysophospholipids cause extracellular K(+) accumulation and action potential shortening with increased risk of arrhythmias during myocardial ischemia. Here we studied effects of several lysophospholipids with different lengths of hydrocarbon chains and charged headgroups on HERG K(+) currents (I(HERG)) expressed in HEK293 cells and the potential mechanisms using whole-cell patch-clamp techniques. 2. Only the lipids with 16 hydrocarbons such as 1-palmitoyl-lysophosphatidylcholine (LPC-16) and 1-palmitoyl lysophosphatidylglycerol (LPG-16) were found to produce significant enhancement of I(HERG) and negative shifts of HERG activation, although the voltage dependence of the effects was different between LPC-16 and LPG-16 which have differently charged headgroups. The lipid with 18 hydrocarbons modestly increased I(HERG). The lipids with 6 or 24 hydrocarbons had no effect or slightly decreased I(HERG). 3. Inhibition or activation of protein kinase C did not alter the effects of LPC-16 and LPG-16. Participation of phosphatidylinositol-4,5 bisphosphate in I(HERG) enhancement by LPC-16/LPG-16 was also excluded. 4. Vitamin E augmented the effects of LPC-16/LPG-16 whereas xanthine/xanthine oxidase reduced I(HERG): indicating that LPC-16/LPG-16 produced dual effects on I(HERG): direct enhancement of I(HERG) and indirect suppression via production of superoxide anion. 5. We conclude that enhancement of HERG function by lysophospholipids is specific to the lipids with 16-hydrocarbon chain structure and the pattern of voltage dependence is determined by the polar headgroups. The increase in I(HERG) is best described by direct interactions between lipid molecules and HERG proteins, which is consistent with lack of effects via membrane destabilization or modulation by intracellular signaling pathways. PMID- 14744815 TI - EP4 prostanoid receptor-mediated vasodilatation of human middle cerebral arteries. AB - 1. Dilatation of the cerebral vasculature is recognised to be involved in the pathophysiology of migraine. Furthermore, elevated levels of prostaglandin E(2) (PGE(2)) occur in the blood, plasma and saliva of migraineurs during an attack, suggestive of a contributory role. In the present study, we have characterised the prostanoid receptors involved in the relaxation and contraction of human middle cerebral arteries in vitro. 2. In the presence of indomethacin (3 microm) and the TP receptor antagonist GR32191 (1 microM), PGE(2) was found to relax phenylephrine precontracted cerebral arterial rings in a concentration-dependent manner (mean pEC(50) 8.0+/-0.1, n=5). 3. Establishment of a rank order of potency using the EP(4)>EP(2) agonist 11-deoxy PGE(1), and the EP(2)>EP(4) agonist PGE(1) OH (mean pEC(50) of 7.6+/-0.1 (n=6) and 6.4+/-0.1 (n=4), respectively), suggested the presence of functional EP(4) receptors. Furthermore, the selective EP(2) receptor agonist butaprost at concentrations <1 microM failed to relax the tissues. 4. Blockade of EP(4) receptors with the EP(4) receptor antagonists AH23848 and EP(4)A caused significant rightward displacements in PGE(2) concentration-response curves, exhibiting pA(2) and pK(B) values of 5.7+/-0.1, n=3, and 8.4, n=3, respectively. 5. The IP receptor agonists iloprost and cicaprost relaxed phenylephrine precontracted cerebral arterial rings (mean pEC(50) values 8.3+/-0.1 (n=4) and 8.1+/-0.1 (n=9), respectively). In contrast, the DP and FP receptor agonists PGD(2) and PGF(2 alpha) failed to cause appreciable relaxation or contraction at concentrations of up to 30 microm. In the absence of phenylephrine contraction and GR32191, the TP receptor agonist U46619 caused concentration-dependent contraction of cerebral artery (mean pEC(50) 7.4+/-0.3, n=3). 6. These data demonstrate the presence of prostanoid EP(4) receptors mediating PGE(2) vasodilatation of human middle cerebral artery. IP receptors mediating relaxation and TP receptors mediating contraction were also functionally demonstrated. PMID- 14744816 TI - Kinin B2 receptor is not involved in enalapril-induced apoptosis and regression of hypertrophy in spontaneously hypertensive rat aorta: possible role of B1 receptor. AB - 1. Treatment with enalapril induces smooth muscle cell apoptosis and regression of aortic hypertrophy in spontaneously hypertensive rats (SHRs), whereas combined blockade of angiotensin II AT(1) and AT(2) receptors does not. We postulated that vascular apoptosis with enalapril involves enhanced half-life of bradykinin (BK) and kinin B(2) receptor stimulation. 2. SHR, 11-weeks old, were treated for 4 weeks with enalapril (30 mg kg(-1) day(-1)), Hoe 140 (500 microg kg(-1) day(-1); B(2) receptor antagonist), alone or in combination. Controls received vehicle. 3. The half-life of hypotensive responses to intra-arterial bolus injections of BK were significantly increased in SHR anesthetized after 4 weeks of enalapril, an effect prevented by Hoe 140. The magnitude of BK-induced hypotension was significantly attenuated in all rats treated with Hoe 140. 4. As compared to placebo, enalapril treatment significantly reduced blood pressure (-34+/-2%), aortic hypertrophy (-20+/-3%), hyperplasia (-37+/-5%) and DNA synthesis (-61+/ 8%), while it increased aortic DNA fragmentation by two-fold. Hoe 140 given alone or in combination with enalapril affected none of these parameters. 5. As a possible alternative mechanism, aortae isolated during the second week of enalapril treatment showed a transient upregulation of contractile responses to des-Arg(9)BK (EC(50)<1 nM), which were significantly reduced by [Leu(8)]des Arg(9)BK (10 microM). Moreover, in vitro receptor autoradiography revealed an increase in expression of B(1) and B(2) receptor binding sites by 8-11 days of enalapril treatment. 6. Aortic apoptosis induction and hypertrophy regression with enalapril do not involve kinin B(2) receptors in SHR. Kinins acting via B(1) receptors remains a candidate mechanism. PMID- 14744817 TI - Lymphocyte function antigen-1 mediates leukocyte adhesion and subsequent liver damage in endotoxemic mice. AB - 1. Sepsis is associated with leukocyte activation and recruitment in the liver. We investigated the role of lymphocyte function antigen-1 (LFA-1) in endotoxin induced leukocyte-endothelium interactions, microvascular perfusion failure, hepatocellular injury and apoptosis in the liver by use of gene-targeted mice, blocking antibodies and a synthetic inhibitor of LFA-1 (LFA703). For this purpose, mice were challenged with lipopolysaccharide (LPS)+D-galactosamine (Gal), and intravital microscopy of the liver microcirculation was conducted 6 h later. 2. The number of firmly adherent leukocytes in response to LPS/Gal was reduced by 48% in LFA-1-deficient mice. Moreover, endotoxin-induced increases of apoptosis and enzyme markers of hepatocellular injury were decreased by 64 and 69 90%, respectively, in LFA-1-deficient mice. Furthermore, sinusoidal perfusion was improved in endotoxemic mice lacking LFA-1. 3. A similar protective pattern was observed in endotoxemic mice pretreated with an antibody against LFA-1. Thus, immunoneutralization of LFA-1 reduced endotoxin-induced leukocyte adhesion by 55%, liver enzymes by 64-66% and apoptosis by 42%, in addition to the preservation of microvascular perfusion. 4. Administration of a novel statin derived inhibitor of LFA-1, LFA703, significantly decreased leukocyte adhesion (more than 56%) and the subsequent liver injury in endotoxemic mice. 5. Thus, this study demonstrates a pivotal role of LFA-1 in supporting leukocyte adhesion in the liver. Moreover, interference with LFA-1-mediated leukocyte adhesion protects against endotoxemic liver damage, and may constitute a potential therapeutic strategy in sepsis. PMID- 14744819 TI - Reduction by gabapentin of K+-evoked release of [3H]-glutamate from the caudal trigeminal nucleus of the streptozotocin-treated rat. AB - Recently, we showed that gabapentin can inhibit a facilitatory effect of substance P (SP) on K(+)-evoked glutamate release in rat trigeminal slices (Maneuf et al., 2001), and we have now examined the effect of gabapentin on glutamate release in the trigeminal slice from the streptozotocin (STZ)-treated rat. 1. At 4 weeks following STZ treatment (50 mg kg(-1) i.p.), blood glucose was increased in the majority of cases, compared to the control level. All the treated animals showed a significant degree (P<0.001) of tactile allodynia (assessed using von Frey filaments) that did not appear to correlate with blood glucose levels. 2. In this study, we demonstrated that, after STZ treatment, 30 microM gabapentin reduced K(+)-evoked release of [(3)H]-glutamate in either normal (11 mM) or high (30 mM) glucose conditions by 24 and 22%, respectively. In the normal rat, gabapentin (up to 100 microM) is ordinarily unable to affect release of glutamate from the trigeminal slice. 3. The uptake of glutamate in Sp5C punches from streptozotocin-treated rats was reduced under normal glucose conditions (41.7% of control), whereas high glucose restored uptake to normal (84.7% of control). 4. The addition of 1 microm substance P potentiated the evoked release of glutamate in both normal (40% increase) and high glucose (28%), and this was blocked by gabapentin (30 microM) in both conditions. It is interesting to speculate that this ability of gabapentin to reduce the release of glutamate in the trigeminal nucleus after streptozotocin treatment may be of relevance to the antihyperalgesic-allodynic actions of the drug. PMID- 14744818 TI - Activation of VPAC1 receptors by VIP and PACAP-27 in human bronchial epithelial cells induces CFTR-dependent chloride secretion. AB - 1. In the human airway epithelium, VIP/PACAP receptors are distributed in nerve fibers and in epithelial cells but their role in transepithelial ion transport have not been reported. Here, we show that human bronchial epithelial Calu-3 cells expressed the VPAC(1) receptor subtype which shares similar high affinity for VIP and PACAP-27. 2. The stoichiometric binding parameters characterizing the (125)I-VIP and (125)I-PACAP-27 binding to these receptors were determined. 3. We found that VIP (EC(50) approximately 7.6 nM) and PACAP-27 (EC(50) approximately 10 nM) stimulated glibenclamide-sensitive and DIDS-insensitive iodide efflux in Calu-3 cells. 4. The protein kinase A (PKA) inhibitor, H-89 and the protein kinase C (PKC) inhibitor, chelerythrine chloride prevented activation by both peptides demonstrating that PKA and PKC are part of the signaling pathway. This profile corresponds to the pharmacological signature of CFTR. 5. In the cystic fibrosis airway epithelial IB3-1 cell lacking functional CFTR but expressing VPAC(1) receptors, neither VIP, PACAP-27 nor forskolin stimulated chloride transport. 6. Ussing chamber experiments demonstrated stimulation of CFTR dependent short-circuit currents by VIP or PACAP-27 applied to the basolateral but not to the apical side of Calu-3 cells monolayers. 7. This study shows the stimulation in human bronchial epithelial cells of CFTR-dependent chloride secretion following activation by VIP and PACAP-27 of basolateral VPAC(1) receptors. PMID- 14744820 TI - Role of oxidative stress and nitric oxide in regulation of spontaneous tone in aorta of DOCA-salt hypertensive rats. AB - 1. The roles of nitric oxide (NO), superoxide anion (O(2)(-)), and hydrogen peroxide (H(2)O(2)) in the modulation of spontaneous tone were investigated in isolated aorta from deoxycorticosterone acetate (DOCA)-salt hypertensive rats. 2. Increases in preload from 1 to 5 g were accompanied by increases in spontaneous tone in aortic rings from DOCA-salt hypertensive rats but not from SHAM normotensive rats. 3. Tone was higher in endothelium-denuded aortic rings than in endothelium-intact vessels. Inhibition of nitric oxide synthase (NOS) with 300 microM N(G)-nitro-L-arginine methyl ester (l-NAME) increased spontaneous tone. 4. Basal O(2)(-) generation was higher in aortic rings from DOCA-salt hypertensive rats than in those from SHAM-normotensive rats. Stretch increased O(2)(-) levels even further in the DOCA-salt group. In rings isolated from DOCA-salt hypertensive rats, administration of the O(2)(-) scavenger, superoxide dismutase (SOD, 150 U ml(-1)), or the nicotinamide adenine dinucleotide phosphate (NADPH) oxidase inhibitor, apocynin (100 microM), completely abolished the development of spontaneous tone in endothelium-intact aortic rings but not in endothelium denuded or in L-NAME-treated rings. SOD and apocynin decreased the generation of O(2)(-) in endothelium-intact, endothelium-denuded, and L-NAME-treated aortic rings. 5. Oral treatment of DOCA-salt hypertensive rats with the O(2)(-) scavengers, tempol or tiron, or with apocynin for 3 weeks prevented the development of hypertension and abolished the increases in O(2)(-) generation and spontaneous tone. 6. Administration of catalase (1000 U ml(-1)) to aortic rings increased spontaneous tone in vessels from DOCA-salt hypertensive rats. 7. Administration of the cyclooxygenase (COX) inhibitor, valeroyl salicylate, or the thromboxane/prostaglandin antagonist, SQ 29548, to aortic rings abolished tone. 8. The results suggest that NO plays a major role in preventing the generation of spontaneous tone in isolated aortic rings from DOCA-salt hypertensive rats. NADPH oxidase-derived O(2)(-) enhanced spontaneous tone by inactivating NO. Endogenous H(2)O(2) appears to mitigate the increase in tone. In addition, a COX component may also contribute to spontaneous tone. PMID- 14744821 TI - Randomised controlled trial of effect of hands and knees posturing on incidence of occiput posterior position at birth. AB - OBJECTIVE: To evaluate the efficacy of hands and knees position and pelvic rocking exercises on the incidence of fetal occiput posterior position at birth. DESIGN: Multicentre randomised controlled trial. SETTING: Seven maternity units in New South Wales, Australia, encompassing teaching hospitals and district general hospitals. PARTICIPANTS: 2547 pregnant women at 37 weeks' gestation; 1292 randomised to the intervention group and 1255 to the control group. INTERVENTION: Hands and knees position and pelvic rocking exercises from 37 weeks' gestation until the onset of labour. MAIN OUTCOME MEASURE: Incidence of fetal occiput posterior position at birth. RESULTS: 1046 women in the intervention group and 1209 women in the control group remained in the study until they went into labour. No significant difference existed between the groups for the incidence of occiput posterior position at birth: 105 (8.1%) women in the intervention group and 98 (7.8%) in the control group had a baby in a posterior position at delivery (difference in risk 0.3%, 95% confidence interval -1.8 to 2.4). The incidence of fetal transverse arrest was 3.4% (44 women) in the intervention group and 3.0% (38 women) in the control group (difference in risk 0.4, -1 to 1.7). No differences occurred between intervention and control groups for induction of labour, use of epidural, duration of labour, mode of delivery, use of episiotomy, or Apgar score. CONCLUSION: Hands and knees exercise with pelvic rocking from 37 weeks' gestation to the onset of labour did not reduce the incidence of persistent occiput posterior position at birth. PMID- 14744822 TI - Randomised controlled trial of labouring in water compared with standard of augmentation for management of dystocia in first stage of labour. AB - OBJECTIVES: To evaluate the impact of labouring in water during first stage of labour on rates of epidural analgesia and operative delivery in nulliparous women with dystocia. DESIGN: Randomised controlled trial. SETTING: University teaching hospital in southern England. PARTICIPANTS: 99 nulliparous women with dystocia (cervical dilation rate < 1 cm/hour in active labour) at low risk of complications. Interventions Immersion in water in birth pool or standard augmentation for dystocia (amniotomy and intravenous oxytocin). MAIN OUTCOME MEASURES: Primary: epidural analgesia and operative delivery rates. Secondary: augmentation rates with amniotomy and oxytocin, length of labour, maternal and neonatal morbidity including infections, maternal pain score, and maternal satisfaction with care. RESULTS: Women randomised to immersion in water had a lower rate of epidural analgesia than women allocated to augmentation (47% v 66%, relative risk 0.71 (95% confidence interval 0.49 to 1.01), number needed to treat for benefit (NNT) 5). They showed no difference in rates of operative delivery (49% v 50%, 0.98 (0.65 to 1.47), NNT 98), but significantly fewer received augmentation (71% v 96%, 0.74 (0.59 to 0.88), NNT 4) or any form of obstetric intervention (amniotomy, oxytocin, epidural, or operative delivery) (80% v 98%, 0.81 (0.67 to 0.92), NNT 5). More neonates of women in the water group were admitted to the neonatal unit (6 v 0, P = 0.013), but there was no difference in Apgar score, infection rates, or umbilical cord pH. CONCLUSIONS: Labouring in water under midwifery care may be an option for slow progress in labour, reducing the need for obstetric intervention, and offering an alternative pain management strategy. PMID- 14744823 TI - Population based study of early risk of stroke after transient ischaemic attack or minor stroke: implications for public education and organisation of services. AB - OBJECTIVE: To estimate the very early stroke risk after a transient ischaemic attack (TIA) or minor stroke and thereby inform the planning of effective stroke prevention services. DESIGN: Population based prospective cohort study of patients with TIA or stroke. SETTING: Nine general practices in Oxfordshire, England, from April 2002 to April 2003. PARTICIPANTS: All patients who had a TIA (n = 87) or minor stroke (n = 87) during the study period and who presented to medical attention. MAIN OUTCOME MEASURES: Risk of recurrent stroke at seven days, one month, and three months after TIAs and minor strokes. RESULTS: The estimated risk of recurrent stroke was 8.0% (95% confidence interval 2.3% to 13.7%) at seven days, 11.5% (4.8% to 18.2%) at one month, and 17.3% (9.3% to 25.3%) at three months after a TIA. The risks at these three time periods after a minor stroke were 11.5% (4.8% to 11.2%), 15.0% (7.5% to 22.5%), and 18.5% (10.3% to 26.7%). CONCLUSIONS: The early risks of stroke after a TIA or minor stroke are much higher than commonly quoted. More research is needed to determine whether these risks can be reduced by more rapid instigation of preventive treatment. PMID- 14744824 TI - Frailty modelling of testicular cancer incidence using Scandinavian data. AB - The incidence of testicular cancer is highest among young men, and then decreases sharply with age. This points towards a frailty effect, where some men have a much greater risk of testicular cancer than the majority of the male population. Those with the highest risk get cancer, drain the group of individuals at risk, and leave a healthy male population which has approximately zero risk of testicular cancer. This leads to the observed decrease in incidence. We discuss a frailty model, where the frailty is compound-Poisson-distributed. This allows for a non-susceptible group (of zero frailty). The model is successfully applied to incidence data from the Danish and Norwegian registries. It is indicated that there was a decrease in incidence for males born during World War II in both countries. Bootstrap analysis is used to find the degree of variation in the estimates. In the Armitage-Doll multistage model, the estimated number of transitions needed for a cell to become malignant is close to 3 for non-seminomas and 4 for seminomas in both the Danish and Norwegian data. This paper demonstrates that a model including a frailty effect fits the incidence data well and gives interesting results and interpretations, although this is no proof of the effect's truth. PMID- 14744825 TI - Composite likelihood and two-stage estimation in family studies. AB - In this paper register based family studies provide the motivation for linking a two-stage estimation procedure in copula models for multivariate failure time data with a composite likelihood approach. The asymptotic properties of the estimators in both parametric and semi-parametric models are derived, combining the approaches of Parner (2001) and Andersen (2003). The method is mainly studied when the families consist of groups of exchangeable members (e.g. siblings) or members at different levels (e.g. parents and children). The advantages of the proposed method are especially clear in this last case where very flexible modelling is possible. The suggested method is also studied in simulations and found to be efficient compared to maximum likelihood. Finally, the suggested method is applied to a family study of deep venous thromboembolism where it is seen that the association between ages at onset is larger for siblings than for parents or for parents and siblings. PMID- 14744826 TI - Estimating treatment effects in studies of perinatal transmission of HIV. AB - Fetal loss often precludes the ascertainment of infection status in studies of perinatal transmission of HIV. The standard analysis based on liveborn babies can result in biased estimation and invalid inference in the presence of fetal death. This paper focuses on the problem of estimating treatment effects for mother-to child transmission when infection status is unknown for some babies. Minimal data structures for identifiability of parameters are given. Methods using full likelihood and the inverse probability of selection-weighted estimators are suggested. Simulation studies are used to show that these estimators perform well in finite samples. Methods are applied to the data from a clinical trial in Dar es Salaam, Tanzania. To validly estimate the treatment effect using likelihood methods, investigators should make sure that the design includes a mini-study among uninfected mothers and that efforts are made to ascertain the infection status of as many babies lost as possible. The inverse probability weighting methods need precise estimation of the probability of observing infection status. We can further apply our methodology to the study of other vertically transmissible infections which are potentially fatal pre- and perinatally. PMID- 14744827 TI - Semi-parametric ROC regression analysis with placement values. AB - Advances in technology provide new diagnostic tests for early detection of disease. Frequently, these tests have continuous outcomes. One popular method to summarize the accuracy of such a test is the Receiver Operating Characteristic (ROC) curve. Methods for estimating ROC curves have long been available. To examine covariate effects, Pepe (1997, 2000) and Alonzo and Pepe (2002) proposed distribution-free approaches based on a parametric regression model for the ROC curve. Cai and Pepe (2002) extended the parametric ROC regression model by allowing an arbitrary non-parametric baseline function. In this paper, while we follow the same semi-parametric setting as in that paper, we highlight a new estimator that offers several improvements over the earlier work: superior efficiency, the ability to estimate the covariate effects without estimating the non-parametric baseline function and easy implementation with standard software. The methodology is applied to a case control dataset where we evaluate the accuracy of the prostate-specific antigen as a biomarker for early detection of prostate cancer. Simulation studies suggest that the new estimator under the semi parametric model, while always being more robust, has efficiency that is comparable to or better than the Alonzo and Pepe (2002) estimator from the parametric model. PMID- 14744828 TI - A comparison of non-iterative and iterative estimators of heterogeneity variance for the standardized mortality ratio. AB - This paper continues work presented in Bohning et al. (2002b, Annals of the Institute of Statistical Mathematics 54, 827-839, henceforth BMSRB) where a class of non-iterative estimators of the variance of the heterogeneity distribution for the standardized mortality ratio was discussed. Here, these estimators are further investigated by means of a simulation study. In addition, iterative estimators including the Clayton-Kaldor procedure as well as the pseudo-maximum likelihood (PML) approach are added in the comparison. Among all candidates, the PML estimator often has the smallest mean square error, followed by the non iterative estimator where the weights are proportional to the external expected counts. This confirms the theoretical result in BMSRB in which an asymptotic efficiency could be proved for this estimator (in the class of non-iterative estimators considered). Surprisingly, the Clayton-Kaldor iterative estimator (often recommended and used by practitioners) performed poorly with respect to the MSE. Given the widespread use of these estimators in disease mapping, medical surveillance, meta-analysis and other areas of public health, the results of this study might be of considerable interest. PMID- 14744829 TI - Nonparametric correction for covariate measurement error in a stratified Cox model. AB - Stratified Cox regression models with large number of strata and small stratum size are useful in many settings, including matched case-control family studies. In the presence of measurement error in covariates and a large number of strata, we show that extensions of existing methods fail either to reduce the bias or to correct the bias under nonsymmetric distributions of the true covariate or the error term. We propose a nonparametric correction method for the estimation of regression coefficients, and show that the estimators are asymptotically consistent for the true parameters. Small sample properties are evaluated in a simulation study. The method is illustrated with an analysis of Framingham data. PMID- 14744830 TI - Factorial and time course designs for cDNA microarray experiments. AB - Microarrays are powerful tools for surveying the expression levels of many thousands of genes simultaneously. They belong to the new genomics technologies which have important applications in the biological, agricultural and pharmaceutical sciences. There are myriad sources of uncertainty in microarray experiments, and rigorous experimental design is essential for fully realizing the potential of these valuable resources. Two questions frequently asked by biologists on the brink of conducting cDNA or two-colour, spotted microarray experiments are 'Which mRNA samples should be competitively hybridized together on the same slide?' and 'How many times should each slide be replicated?' Early experience has shown that whilst the field of classical experimental design has much to offer this emerging multi-disciplinary area, new approaches which accommodate features specific to the microarray context are needed. In this paper, we propose optimal designs for factorial and time course experiments, which are special designs arising quite frequently in microarray experimentation. Our criterion for optimality is statistical efficiency based on a new notion of admissible designs; our approach enables efficient designs to be selected subject to the information available on the effects of most interest to biologists, the number of arrays available for the experiment, and other resource or practical constraints, including limitations on the amount of mRNA probe. We show that our designs are superior to both the popular reference designs, which are highly inefficient, and to designs incorporating all possible direct pairwise comparisons. Moreover, our proposed designs represent a substantial practical improvement over classical experimental designs which work in terms of standard interactions and main effects. The latter do not provide a basis for meaningful inference on the effects of most interest to biologists, nor make the most efficient use of valuable and limited resources. PMID- 14744831 TI - Quantifying and comparing the predictive accuracy of continuous prognostic factors for binary outcomes. AB - The positive and negative predictive values are standard ways of quantifying predictive accuracy when both the outcome and the prognostic factor are binary. Methods for comparing the predictive values of two or more binary factors have been discussed previously (Leisenring et al., 2000, Biometrics 56, 345-351). We propose extending the standard definitions of the predictive values to accommodate prognostic factors that are measured on a continuous scale and suggest a corresponding graphical method to summarize predictive accuracy. Drawing on the work of Leisenring et al. we make use of a marginal regression framework and discuss methods for estimating these predictive value functions and their differences within this framework. The methods presented in this paper have the potential to be useful in a number of areas including the design of clinical trials and health policy analysis. PMID- 14744832 TI - Tests for multivariate recurrent events in the presence of a terminal event. AB - In studies involving diseases associated with high rates of mortality, trials are frequently conducted to evaluate the effects of therapeutic interventions on recurrent event processes terminated by death. In this setting, cumulative mean functions form a natural basis for inference for questions of a health economic nature, and Ghosh and Lin (2000) recently proposed a relevant class of test statistics. Trials of patients with cancer metastatic to bone, however, involve multiple types of skeletal complications, each of which may be repeatedly experienced by patients over their lifetime. Traditionally the distinction between the various types of events is ignored and univariate analyses are conducted based on a composite recurrent event. However, when the events have different impacts on patients' quality of life, or when they incur different costs, it can be important to gain insight into the relative frequency of the specific types of events and treatment effects thereon. This may be achieved by conducting separate marginal analyses with each analysis focusing on one type of recurrent event. Global inferences regarding treatment benefit can then be achieved by carrying out multiplicity adjusted marginal tests, more formal multiple testing procedures, or by constructing global test statistics. We describe methods for testing for differences in mean functions between treatment groups which accommodate the fact that each particular event process is ultimately terminated by death. The methods are illustrated by application to a motivating study designed to examine the effect of bisphosphonate therapy on the incidence of skeletal complications among patients with breast cancer metastatic to bone. We find that there is a consistent trend towards a reduction in the cumulative mean for all four types of skeletal complications with bisphosphonate therapy; there is a significant reduction in the need for radiation therapy for the treatment of bone. The global test suggests that bisphosphonate therapy significantly reduces the overall number of skeletal complications. PMID- 14744833 TI - How special is a 'special' interval: modeling departure from length-biased sampling in renewal processes. AB - Length-biased sampling occurs in renewal processes when the probability that an interval is selected is proportional to the length of the interval. This can occur when intervals are selected because they contain an event that is independent of the renewal process and occurs with constant hazard. For example, if the times between donations for repeat blood donors are independent and identically distributed, and if the donor seroconverts to HIV (develops antibodies that indicate infection with human immunodeficiency virus), then the interval between the last HIV seronegative and first HIV seropositive test is expected to be longer than that donor's previous time intervals between donations. We develop hypothesis tests to determine if the relationship between the typical and length-biased intervals is as expected, or if there is departure from length-biased sampling. We further develop a regression method to determine if there are covariates that explain the departure from length-biased sampling. Our approach is motivated by the question of whether there is evidence that repeat blood donors who develop antibodies to HIV or other viral infections change their donation pattern in some way because of seroconversion. PMID- 14744834 TI - Treatment of the patient with diabetes mellitus and risk of nephropathy: what do we know, and what do we need to learn? PMID- 14744836 TI - Updates on the treatment of epilepsy in women. AB - Epilepsy is a condition of the central nervous system that is characterized by recurrent seizures. The goal of management is to make patients seizure free without intolerable adverse effects from treatment. Men and women differ in their physiologic makeup and therefore have different needs that must be considered when attempting to attain this goal. There are special concerns for women of child-bearing years with regard to contraception, pregnancy, and teratogenicity that should be considered during counseling and selection of appropriate treatment. There are also emerging concerns about the interaction of antiepileptic drugs and endocrine function that can affect ovarian function, induce polycystic ovary (PCO)-like syndrome, and threaten fertility. Systemic adverse effects can have a negative impact on weight, cosmetic appearance, sexual function, and bone health. Individualized treatment coupling antiepileptic drug use and the specific phase of impact of the reproductive cycle must be considered in treatment selection. Important concerns regarding long-term therapy are being raised as there are more treatment options to consider because of the plethora of new antiepileptic drugs that are available, often with more favorable pharmacokinetics and different adverse event profiles. Also, sex hormone fluctuations during maturation may exacerbate seizures at particular points during the life cycle for women, including menarche, during menses, during pregnancy, or later in the perimenopausal years, often presenting a uniquely challenging aspect to treatment. As the number of available treatment options for epilepsy increases, the optimal goal for primary care physicians is to work as a team with obstetricians, gynecologists, and neurologists in an effort to ensure the best treatment of women with epilepsy. PMID- 14744837 TI - Use of statins and fracture: results of 4 prospective studies and cumulative meta analysis of observational studies and controlled trials. AB - BACKGROUND: The 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors (statins) are widely used for the treatment of hyperlipidemia, and recent in vitro and animal data suggest that statins promote bone formation and increase bone strength. METHODS: To determine whether statin use is associated with a reduced risk for fracture, we analyzed statin use and fracture rates in 4 large prospective studies (the Study of Osteoporotic Fractures, the Fracture Intervention Trial, the Heart and Estrogen/Progestin Replacement Study, and the Rotterdam Study). We searched MEDLINE through January 2002 and abstracts from major scientific meetings and performed a cumulative meta-analysis of published and unpublished observational studies and clinical trials. The meta-analysis included 8 observational studies and 2 clinical trials that reported statin use and documented fracture outcomes. RESULTS: After adjustment for multiple factors, including age, body mass index, and estrogen use, we found a trend toward fewer hip fractures (relative hazards [RHs], 0.19-0.62) and, to a lesser extent, nonspine fractures (RHs, 0.49-0.95) among statin users in each of the 4 prospective studies. The meta-analysis of observational studies was consistent with these findings. The summary odds ratio (OR) for statin use and hip fracture was 0.43 (95% confidence interval [CI], 0.25-0.75), whereas that for nonspine fracture was 0.69 (95% CI, 0.55-0.88). The meta-analysis of clinical trial results did not support a protective effect with statin use for hip fracture (summary OR, 0.87; 95% CI, 0.48-1.58) or nonspine fracture (OR, 1.02; 95% CI, 0.83-1.26). CONCLUSIONS: Observational studies suggest that the risk for hip and nonspine fractures is lower among older women taking statin medications for hyperlipidemia, but post hoc analyses of cardiovascular trials do not. Controlled trials specifically designed to test the effect of statins on skeletal metabolism and fracture are needed. PMID- 14744839 TI - A twin study of erectile dysfunction. AB - BACKGROUND: The extent of genetic influence on erectile dysfunction (ED) is unknown. This study determines the contribution of heredity to ED in a sample of middle-aged men. METHODS: A classical twin study was conducted in the Vietnam Era Twin Registry, a national sample of male-male pairs (mean birth year, 1949) who served on active duty during the Vietnam era (1965-1975). A 1999 male health survey was completed by 890 monozygotic (MZ) and 619 dizygotic (DZ) pairs. The prevalence and heritability of 2 self-report indicators of ED, difficulty in having an erection and in maintaining an erection, are estimated. RESULTS: The prevalence of difficulty in having an erection is 23.3% and in maintaining an erection is 26.7%. Twin correlations for dysfunction in having an erection are 0.35 (95% confidence interval [CI], 0.28-0.41) in MZ and 0.17 (95% CI, 0.09-0.27) in DZ pairs. For dysfunction in maintaining an erection, the twin correlations in MZ and DZ pairs are 0.39 (95% CI, 0.32-0.45) and 0.18 (95% CI, 0.09-0.27), respectively. The estimated heritability of liability for dysfunction in having an erection is 35% and in maintaining an erection is 42%. The heritable influence on ED remained significant after adjustment for ED risk factors. CONCLUSIONS: The present study demonstrates an ED-specific genetic component that is independent of genetic influences from numerous ED risk factors. The results suggest that future molecular genetic studies to identify ED-related polymorphisms are warranted. PMID- 14744840 TI - Accuracy of ultrasonography in predicting celiac disease. AB - BACKGROUND: Various ultrasonographic (US) signs have been reported in overt celiac disease (CD). The aim of this study was to investigate the diagnostic accuracy of 6 US parameters in predicting CD. METHODS: One hundred sixty-two consecutive patients with chronic diarrhea (n=105), iron deficiency anemia (n=25), or dyspepsia (n=32) underwent anti-endomysial IgA antibody determination and duodenal biopsy. Moreover, US evaluation of 6 parameters (ie, fasting gallbladder volume, transverse diameter of small bowel loops, thickness of the small bowel wall, pattern of peristalsis, presence of free abdominal fluid, and diameter of the mesenteric lymph nodes) was done by 2 operators blind to the serological and histological findings. The pretest probability of CD was estimated to be between 5% and 10%. The percentage of agreement between US and histologic findings, the sensitivity, specificity, positive and negative likelihood ratios, and the posttest probability for positive and negative results were calculated. RESULTS: Celiac disease was diagnosed in 12 patients (7.4%). An increased gallbladder volume, the presence of free fluid in the abdominal cavity, and enlarged mesenteric lymph nodes showed a specificity of 96%, 96%, and 97%, respectively (95% confidence intervals [CIs], 92%-99%, 93%-99%, and 95%-99%), whereas the presence of dilated small bowel loops with increased fluid content and increased peristalsis had a sensitivity of 92% and 83%, respectively (95% CIs, 76%-100% and 62%-100%). Eleven (92%) of the 12 patients with celiac disease and 35 (23%) of the 150 patients who did not have the disease had at least 1 US sign (P=.001); all of the US signs were concomitantly present in 4 patients with CD (33%) and 1 patient without CD (0.6%) (P=.001). CONCLUSION: Ultrasonographic evaluation can accurately predict CD but its place in the diagnostic algorithm depends upon the probability of the disease in the considered population. PMID- 14744841 TI - Pain and satisfaction with pain control in hospitalized medical patients: no such thing as low risk. AB - BACKGROUND: Pain is common in many populations of hospitalized patients. It is unknown if any populations of hospitalized patients are at low risk for pain. We studied the prevalence of pain and satisfaction with pain control in a general medicine inpatient service to determine if this population was at low risk for pain. METHODS: We performed a prospective cohort study of 5584 hospitalized patients. Pain and pain control were assessed in a follow-up telephone survey. Predictors of pain were determined through administrative databases and patient survey. RESULTS: Of the study patients, 59% had pain (28% reported severe, 19% moderate, and 12% mild pain). Among patients with common diagnoses, those with sickle cell crisis were the most likely and those with syncope were the least likely to report significant pain (90% and 34%, respectively). Patient characteristics significantly associated with increased pain included DRG (diagnosis related group) weight (odds ratio [OR], 1.19), Charlson Index score (OR, 1.03), age older than 65 years (OR, 0.65), female sex (OR, 1.17), and education level higher than high school (OR, 1.31). Pain was reported by 28% of patients without high-risk characteristics for pain; and 82.2% of patients were satisfied, 11.1% somewhat satisfied, and 6.7% dissatisfied with their pain treatment. CONCLUSIONS: Pain was common in the study population and more patients rated their pain as severe than as moderate or mild. Pain was dispersed among diagnoses. Although most patients thought that their pain was adequately controlled, 18% of patients with pain (10% of all patients) reported that their pain was inadequately controlled. Although patient characteristics were associated with pain or dissatisfaction with pain control, they were weak predictors and significant pain was common even in populations at the lowest-risk for pain. PMID- 14744842 TI - Trends in self-reported multiple cardiovascular disease risk factors among adults in the United States, 1991-1999. AB - BACKGROUND: There are few national- and state-level data on multiple cardiovascular disease (CVD) risk factor status and trends over time. We examined the prevalence of self-reported multiple CVD risk factors from 1991 through 1999. METHODS: The Behavioral Risk Factor Surveillance System is a state-based telephone survey of adults 18 years or older. Surveys in 1991, 1993, 1995, 1997, and 1999 ascertained reported high blood pressure, high blood cholesterol level, diabetes, obesity, and current smoking status. Trends in the prevalence of persons with each risk factor and of having 2 or more risk factors were calculated. Data were age standardized to the 2000 US population. RESULTS: From 1991 to 1999, the prevalence of reported high blood pressure increased from 23.8% to 25.4%, high cholesterol levels increased from 24.9% to 27.7%, diabetes increased from 5.5% to 7.1%, obesity increased from 13.5% to 20.3%, and smoking remained at approximately 21%. The prevalence of adults with 2 or more risk factors increased from 23.6% in 1991 to 27.9% in 1999 and significantly increased for both men and women and for all race or ethnic, age, and education groups. Among states, the prevalence of multiple risk factors ranged from 15.0% to 29.9% in 1991 and from 18.7% to 37.1% in 1999. From 1991 to 1999, the prevalence of multiple risk factors increased by 10% or more in 36 states. CONCLUSIONS: The substantial proportion of persons with known multiple risk factors (25% of the population) suggests that increased CVD prevention and risk factor reduction efforts should focus on comprehensive risk reduction strategies. PMID- 14744838 TI - Conceptual foundations of the UCSD Statin Study: a randomized controlled trial assessing the impact of statins on cognition, behavior, and biochemistry. AB - BACKGROUND: Statin cholesterol-lowering drugs are among the most prescribed drugs in the United States. Their cardiac benefits are substantial and well supported. However, there has been persistent controversy regarding possible favorable or adverse effects of statins or of cholesterol reduction on cognition, mood, and behavior (including aggressive or violent behavior). METHODS: The literature pertaining to the relationship of cholesterol or statins to several noncardiac domains was reviewed, including the link between statins (or cholesterol) and cognition, aggression, and serotonin. RESULTS: There are reasons to think both favorable and adverse effects of statins and low cholesterol on cognition may pertain; the balance of these factors requires further elucidation. A substantial body of literature links low cholesterol level to aggressive behavior; statin randomized trials have not supported a connection, but they have not been designed to address this issue. A limited number of reports suggest a connection between reduced cholesterol level and reduced serotonin level, but more information is needed with serotonin measures that are practical for clinical use. Whether lipophilic and hydrophilic statins differ in their impact should be assessed. CONCLUSION: There is a strong need for randomized controlled trial data to more clearly establish the impact of hydrophilic and lipophilic statins on cognition, aggression, and serotonin, as well as on other measures relevant to risks and quality-of-life impact in noncardiac domains. PMID- 14744843 TI - Symptomatic venous thromboembolism in cancer patients treated with chemotherapy: an underestimated phenomenon. AB - BACKGROUND: The exact incidence of venous thromboembolism (VTE) in cancer patients is unknown, partly because of confounding factors. Prophylactic treatment is warranted in surgical patients with cancer because of a high incidence of VTE. We performed a retrospective study to evaluate if the same applies for cancer patients treated with chemotherapy. METHODS: The medical records of 206 consecutive patients with malignancy, treated with chemotherapy, were identified. The kind of malignancy and chemotherapeutic treatment were recorded, as was the date of treatment. The records were reviewed for other risk factors for VTE, and were searched for proved deep venous thrombosis or pulmonary embolism. RESULTS: Of those 206 patients, 15 (7.3%) had proved VTE during or within 3 months after chemotherapeutic treatment. The annual incidence was 10.9%. The incidence of VTE was specifically high in the 39 patients treated with a combination of fluorouracil and leucovorin calcium because of colorectal cancer (6 [15%] of the patients were affected). The occurrence of VTE in the latter group of patients was not influenced by factors such as surgery, central venous catheters, or tumor load. CONCLUSIONS: The annual incidence of VTE in patients treated with chemotherapy was high, specifically in patients with colorectal cancer treated with fluorouracil-leucovorin. If these observations are confirmed, trials to evaluate the use of prophylactic anticoagulant treatment should be conducted. PMID- 14744844 TI - Sex and the risk of restless legs syndrome in the general population. AB - BACKGROUND: Restless legs syndrome (RLS) is characterized by the desire to move the limbs associated with paresthesias of the legs, a motor restlessness, an intensification of symptoms at rest with relief by activity, and a worsening of symptoms in the evening or at night. Population-based studies are rare, and risk factors in the general population are not known. METHODS: Cross-sectional survey with face-to-face interviews and physical examination among 4310 participants in the Study of Health in Pomerania in northeastern Germany. Participants were aged 20 to 79 years and were randomly selected from population registers. Restless legs syndrome was assessed with standardized, validated questions addressing the 4 minimal criteria for RLS as defined by the International Restless Legs Syndrome Study Group. RESULTS: The overall prevalence of RLS was 10.6%, increasing with age, and women were twice as often affected as men. While nulliparous women had prevalences similar to those among men up to age 64 years, the risk of RLS increased gradually for women with 1 child (odds ratio, 1.98; 95% confidence interval, 1.25-3.13), 2 children (odds ratio, 3.04; 95% confidence interval, 2.11 4.40), and 3 or more children (odds ratio, 3.57; 95% confidence interval, 2.30 5.55). Subjects with RLS had significantly lower quality-of-life scores than those without the syndrome. CONCLUSIONS: Restless legs syndrome is a common disease in the general population, affecting women more often than men. It is associated with reduced quality of life in cross-sectional analysis. Parity is a major factor in explaining the sex difference and may guide further clarification of the etiology of the disease. PMID- 14744845 TI - Get with the guidelines for cardiovascular secondary prevention: pilot results. AB - BACKGROUND: The use of Web-based technology and a collaborative model to improve hospital adherence to secondary prevention guidelines has not been previously evaluated. METHODS: Twenty-four hospitals in Massachusetts participated in a collaborative that met quarterly, with didactic and best-practice presentations and interactive multidisciplinary team workshops. A customized tool kit and interactive, Web-based management tool were used for data collection and on-line feedback. Data from 1738 patients admitted with coronary artery disease were collected by hospital staff from July 1, 2000, to June 30, 2001. Outcome measures included differences between baseline and 10- to 12-month follow-up measurements of use of aspirin, beta-blockers, angiotensin-converting enzyme inhibitors, cholesterol measurement and treatment, smoking cessation counseling, blood pressure control, and cardiac rehabilitation referral. RESULTS: Clinically and statistically significant increases from baseline to 10- to 12-month follow-up were demonstrated in smoking cessation counseling (48% [95% confidence interval [CI], 36.6%-58.4%] to 87% [95% CI, 73.1%-100.7%]), lipid treatment (54% [95% CI, 46.6%-70.2%] to 79% [95% CI, 70.2%-88.3%]), lipid measurement (59% [95% CI, 51.5% 66.0%] to 81% [95% CI, 72.0%-89.5%]), and cardiac rehabilitation referral (34% [95% CI, 25.9%-39.7%] to 73% [95% CI, 63.2%-82.9%]). An improving trend was seen in blood pressure control (60% [95% CI, 55.3%-65.6%] to 68% [95% CI, 60.2% 76.1%]). High baseline use was maintained for use of aspirin, beta-blockers, and angiotensin-converting enzyme inhibitors. CONCLUSION: Implementation of a collaborative quality improvement initiative, interactive training of hospital teams with physician champions, and the use of an interactive Web-based Patient Management Tool enhanced adherence to prevention guidelines in hospitalized patients with coronary artery disease. PMID- 14744846 TI - Effects of an ad libitum low-fat, high-carbohydrate diet on body weight, body composition, and fat distribution in older men and women: a randomized controlled trial. AB - BACKGROUND: The efficacy of ad libitum low-fat diets in reducing body weight and fat in overweight and obese adults remains controversial. METHODS: We examined the effect of a 12-week low-fat, high-complex carbohydrate diet alone (HI-CHO) and in combination with aerobic exercise training (HI-CHO + EX) on body weight and composition in 34 individuals with impaired glucose tolerance (20 women and 14 men; mean +/- SEM age, 66 +/- 1 years). Participants were randomly assigned to a control diet (41% fat, 14% protein, 45% carbohydrates, and 7 g of fiber per 1000 kcal), a HI-CHO diet (18% fat, 19% protein, 63% carbohydrates, and 26 g of fiber per 1000 kcal), or a HI-CHO diet plus endurance exercise 4 d/wk, 45 min/d, at 80% peak oxygen consumption (HI-CHO + EX). Participants were provided 150% of estimated energy needs and were instructed to consume food ad libitum. Total food intake, body composition, resting metabolic rate, and substrate oxidation were measured. RESULTS: There was no significant difference in total food intake among the 3 groups and no change in energy intake over time. The HI-CHO + EX and HI-CHO groups lost more body weight (-4.8 +/- 0.9 kg [P=.003] and -3.2 +/- 1.2 kg [P=.02]) and a higher percentage of body fat (-3.5% +/- 0.7% [P=.01] and -2.2% +/ 1.2% [P=.049]) than controls (-0.1 +/- 0.6 kg and 0.2% +/- 0.6%). In addition, thigh fat area decreased in the HI-CHO (P=.003) and HI-CHO + EX (P<.001) groups compared with controls. High carbohydrate intake and weight loss did not result in a decreased resting metabolic rate or reduced fat oxidation. CONCLUSION: A high-carbohydrate diet consumed ad libitum, with no attempt at energy restriction or change in energy intake, results in losses of body weight and body fat in older men and women. PMID- 14744847 TI - Quinine/quinidine-induced thrombocytopenia: a great imitator. PMID- 14744848 TI - Rocky Mountain spotted fever revisited. PMID- 14744849 TI - In-hospital mortality and long-term use of inhaled corticosteroids. PMID- 14744850 TI - Achieved vs initial blood pressure in predicting renal outcomes. PMID- 14744851 TI - Do findings of high mortality from pneumonia in the elderly make it the old man's friend? PMID- 14744852 TI - Prolonged hypoxia differentially regulates hypoxia-inducible factor (HIF)-1alpha and HIF-2alpha expression in lung epithelial cells: implication of natural antisense HIF-1alpha. AB - Transcriptional adaptations to hypoxia are mediated by hypoxia-inducible factor (HIF)-1, a heterodimer of HIF-alpha and aryl hydrocarbon receptor nuclear translocator subunits. The HIF-1alpha and HIF-2alpha subunits both undergo rapid hypoxia-induced protein stabilization and bind identical target DNA sequences. When coexpressed in similar cell types, discriminating control mechanisms may exist for their regulation, explaining why HIF-1alpha and HIF-2alpha do not substitute during embryogenesis. We report that, in a human lung epithelial cell line (A549), HIF-1alpha and HIF-2alpha proteins were similarly induced by acute hypoxia (4 h, 0.5% O(2)) at the translational or posttranslational level. However, HIF-1alpha and HIF-2alpha were differentially regulated by prolonged hypoxia (12 h, 0.5% O(2)) since HIF-1alpha protein stimulation disappeared because of a reduction in its mRNA stability, whereas HIF-2alpha protein stimulation remained high and stable. Prolonged hypoxia also induced an increase in the quantity of natural antisense HIF-1alpha (aHIF), whose gene promoter contains several putative hypoxia response elements to which (as we confirm here) the HIF-1alpha or HIF-2alpha protein can bind. Finally, transient transfection of A549 cells by dominant-negative HIF-2alpha, also acting as a dominant-negative for HIF-1alpha, prevented both the decrease in the HIF-1alpha protein and the increase in the aHIF transcript. Taken together, these data indicate that, during prolonged hypoxia, HIF-alpha proteins negatively regulate HIF-1alpha expression through an increase in aHIF and destabilization of HIF-1alpha mRNA. This trans regulation between HIF-1alpha and HIF-2alpha during hypoxia likely conveys target gene specificity. PMID- 14744853 TI - A threshold mechanism governing activation of the developmental regulatory protein sigma F in Bacillus subtilis. AB - The RNA polymerase sigma factor sigma(F) is a developmental regulatory protein that is activated in a cell-specific manner following the formation of the polar septum during the process of spore formation in the bacterium Bacillus subtilis. Activation of sigma(F) depends on the membrane-bound phosphatase SpoIIE, which localizes to the septum, and on the formation of the polar septum itself. SpoIIE is responsible for dephosphorylating and thereby activating the phosphoprotein SpoIIAA, which, in turn, triggers the release of sigma(F) from the anti-sigma(F) factor SpoIIAB. Paradoxically, however, the presence of unphosphorylated SpoIIAA is insufficient to cause sigma(F) activation as SpoIIAA reaches substantial levels in mutants blocked in polar septation. We now describe mutants of SpoIIE, SpoIIAA, and SpoIIAB that break the dependence of sigma(F) activation on polar division. Analysis of these mutants indicates that unphosphorylated SpoIIAA must reach a threshold concentration in order to trigger the release of sigma(F) from SpoIIAB. Evidence is presented that this threshold is created by the action of SpoIIAB, which can form an alternative, long lived complex with SpoIIAA. We propose that formation of the SpoIIAA-SpoIIAB complex serves as a sink that traps SpoIIAA in an inactive state and that only when unphosphorylated SpoIIAA is in excess to the sink does activation of sigma(F) take place. PMID- 14744854 TI - Double strand break repair by homologous recombination is regulated by cell cycle independent signaling via ATM in human glioma cells. AB - To investigate double strand break (DSB) repair and signaling in human glioma cells, we stably transfected human U87 (ATM(+), p53(+)) glioma cells with a plasmid having a single I-SceI site within an inactive green fluorescent protein (GFP) expression cassette, allowing for the detection of homologous recombination repair (HRR) by GFP expression. HRR and nonhomologous end joining (NHEJ) were also determined by PCR. DSB repair was first detected at 12 h postinfection with an adenovirus expressing I-SceI with repair reaching plateau levels between 24 and 48 h. Within this time frame, NHEJ predominated over HRR in the range of 3-50 fold. To assess the involvement of ATM in DSB repair, we first examined whether ATM was associated with the DSB. Chromatin immunoprecipitation showed that ATM was present at the site of the DSB as early as 18 h postinfection. In cells treated with caffeine, an inhibitor of ATM, HRR was reduced, whereas NHEJ was not. In support of this finding, GFP flow cytometry demonstrated that caffeine reduced HRR by 90% under conditions when ATM kinase activity was inhibited. Dominant-negative ATM expressed from adenovirus inhibited HRR by 45%, also having little to no effect on NHEJ. Furthermore, HRR was inhibited by caffeine in serum starved cells arrested in G(0)/G(1), suggesting that ATM is also important for HRR outside of the S and G(2) cell cycle phases. Altogether, these results demonstrate that HRR contributes substantially to DSB repair in human glioma cells, and, importantly, ATM plays a critical role in regulating HRR but not NHEJ throughout the cell cycle. PMID- 14744855 TI - Lysophosphatidic acid and autotaxin stimulate cell motility of neoplastic and non neoplastic cells through LPA1. AB - Autotaxin (ATX) is a tumor cell motility-stimulating factor originally isolated from melanoma cell supernatant that has been implicated in regulation of invasive and metastatic properties of cancer cells. Recently, we showed that ATX is identical to lysophospholipase D, which converts lysophosphatidylcholine to a potent bioactive phospholipid mediator, lysophosphatidic acid (LPA), raising the possibility that autocrine or paracrine production of LPA by ATX contributes to tumor cell motility. Here we demonstrate that LPA and ATX mediate cell motility stimulating activity through the LPA receptor, LPA(1). In fibroblasts isolated from lpa(1)(-/-) mice, but not from wild-type or lpa(2)(-/-), cell motility stimulated with LPA and ATX was completely absent. In the lpa(1)(-/-) cells, LPA stimulated lamellipodia formation was markedly diminished with a concomitant decrease in Rac1 activation. LPA stimulated the motility of multiple human cancer cell lines expressing LPA(1), and the motility was attenuated by an LPA(1) selective antagonist, Ki16425. The present study suggests that ATX and LPA(1) represent potential targets for cancer therapy. PMID- 14744856 TI - Mammalian electron transferring flavoprotein.flavoprotein dehydrogenase complexes observed by microelectrospray ionization-mass spectrometry and surface plasmon resonance. AB - Microelectrospray ionization-mass spectrometry was used to directly observe electron transferring flavoprotein.flavoprotein dehydrogenase interactions. When electron transferring flavoprotein and porcine dimethylglycine dehydrogenase or sarcosine dehydrogenase were incubated together in the absence of substrate, a relative molecular mass corresponding to the flavoprotein.electron transferring flavoprotein complex was observed, providing the first direct observation of these mammalian complexes. When an acyl-CoA dehydrogenase family member, human short chain acyl-CoA dehydrogenase, was incubated with dimethylglycine dehydrogenase and electron transferring flavoprotein, the microelectrospray ionization-mass spectrometry signal for the dimethylglycine dehydrogenase.electron transferring flavoprotein complex decreased, indicating that the acyl-CoA dehydrogenases have the ability to compete with the dimethylglycine dehydrogenase/sarcosine dehydrogenase family for access to electron transferring flavoprotein. Surface plasmon resonance solution competition experiments revealed affinity constants of 2.0 and 5.0 microm for the dimethylglycine dehydrogenase-electron transferring flavoprotein and short chain acyl-CoA dehydrogenase-electron transferring flavoprotein interactions, respectively, suggesting the same or closely overlapping binding motif(s) on electron transferring flavoprotein for dehydrogenase interaction. PMID- 14744857 TI - Synthesis in vitro of rabbit hemorrhagic disease virus subgenomic RNA by internal initiation on (-)sense genomic RNA: mapping of a subgenomic promoter. AB - Rabbit hemorrhagic disease virus (RHDV), a positive-strand RNA virus, is the type species of the Lagovirus within the Caliciviridae. In addition to the genomic RNA of 7.4 kb, a subgenomic mRNA (sgRNA) of 2.2 kb, which is identical in sequence to the 3' one-third of the genomic RNA, is also synthesized in RHDV-infected cells. Numerous RNA viruses make sgRNA for expression of their 3'-proximal genes. A relevant mechanism for viral gene expression is the regulation of sgRNA synthesis by specific promoter elements. In this study, we have investigated in vitro the sgRNA synthesis mechanism using recombinant RHDV RNA-dependent RNA polymerase produced in baculovirus-infected insect cells and synthetic RHDV (-)RNAs of different lengths containing regions located upstream of the subgenomic start site. We report evidences supporting that the sgRNA of RHDV is synthesized in vitro by internal initiation (subgenomic promoter) on (-)RNA templates of genomic length. The deletion mapping of the subgenomic promoter starting from minus strand genomic length RNA showed that a sequence of 50 nucleotides upstream of the sgRNA start site (+1) is sufficient for full subgenomic promoter activity in an in vitro assay using recombinant RHDV RNA-dependent RNA polymerase. This study reports the first description of a subgenomic promoter in a member of the Caliciviridae. PMID- 14744858 TI - Primary structure and in vitro antibacterial properties of the Drosophila melanogaster attacin C Pro-domain. AB - In Drosophila melanogaster, seven distinct families of antimicrobial peptides with different structures and specificities are synthesized by the fat body and released into the hemolymph during the immune response. Using microscale high performance liquid chromatography, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, and Edman degradation, we have isolated and characterized from immune-challenged Drosophila two novel induced molecules, under the control of the Imd pathway, that correspond to post-translationally modified antimicrobial peptides or peptide fragments. The first molecule is a doubly glycosylated form of drosocin, an O-glycosylated peptide that kills Gram negative organisms. The second molecule represents a truncated form of the pro domain of the Drosophila attacin C carrying two post-translational modifications and has significant structural similarities to proline-rich antibacterial peptides including drosocin. We have synthesized this peptide and found that it is active against Gram-negative bacteria. Furthermore, this activity is potentiated when the peptide is used in combination with the Drosophila antimicrobial peptide cecropin A. The synergistic action observed between these two molecules suggests that the truncated post-translationally modified pro domain of attacin C by itself may play an important role in the antimicrobial defense of Drosophila. PMID- 14744859 TI - Human mitotic spindle-associated protein PRC1 inhibits MgcRacGAP activity toward Cdc42 during the metaphase. AB - Although many proteins have been shown to participate in mitotic events, including cytokinesis, their specific roles and interactions remain unclear. A novel interaction of proteins is demonstrated in this report. Yeast two-hybrid screening using PRC1 (protein-regulating cytokinesis 1) cDNA, a human mitotic spindle-associated cyclin-dependent kinase (CDK) substrate, which is involved in cytokinesis, as bait was performed. Data show that the PRC1 bait bound to MgcRacGAP, which is a GTPase-activating protein (GAP) for the Rho family GTPases also involved in cytokinesis. In addition, the two proteins showed similar localization during the M phase. PRC1 was shown to bind to the COOH-terminal GAP conserved domain of MgcRacGAP and to inhibit its GAP activity toward Cdc42. This binding and/or inhibition of MgcRacGAP GAP activity was found to depend on further binding of PRC1 to the basic region (125-285 amino acids) of MgcRacGAP. Furthermore, the basic region was phosphorylated with Aurora B kinase, and this phosphorylation prevented the inhibition of GAP activity by PRC1. Cells overexpressing a phosphorylation mimic mutant of MgcRacGAP exhibited an abnormality of spindle morphology in the metaphase. Cdc42 showed high activity and was localized to the mitotic spindles and centrosomes during the metaphase. We propose that PRC1 down-regulates the GAP activity of MgcRac-GAP during the metaphase and thereby contributes to the correct formation of the spindle. PMID- 14744860 TI - Ribosome stalling during translation elongation induces cleavage of mRNA being translated in Escherichia coli. AB - Recently, it has been found that ribosome pausing at stop codons caused by certain nascent peptides induces cleavage of mRNA in Escherichia coli cells (1, 2). The question we addressed in the present study is whether mRNA cleavage occurs when translation elongation is prevented. We focused on a specific peptide sequence (AS17), derived from SecM, that is known to cause elongation arrest. When the crp-crr fusion gene encoding CRP-AS17-IIA(Glc) was expressed, cAMP receptor protein (CRP) proteins truncated around the arrest sequence were efficiently produced, and they were tagged by the transfer-messenger RNA (tmRNA) system. Northern blot analysis revealed that both truncated upstream crp and downstream crr mRNAs were generated along with reduced amounts of the full-length crp-crr mRNA. The truncated crp mRNA dramatically decreased in the presence of tmRNA due to rapid degradation. The 3' ends of truncated crp mRNA correspond well to the C termini of the truncated CRP proteins. We conclude that ribosome stalling by the arrest sequence induces mRNA cleavage near the arrest point, resulting in nonstop mRNAs that are recognized by tmRNA. We propose that the mRNA cleavage induced by ribosome stalling acts in concert with the tmRNA system as a way to ensure quality control of protein synthesis and possibly to regulate the expression of certain genes. PMID- 14744861 TI - Proprotein convertase furin interacts with and cleaves pro-ADAMTS4 (Aggrecanase 1) in the trans-Golgi network. AB - A member of the A disintegrin and metalloproteinase domain with thrombospondin type-1 motifs (ADAMTS-4) protease family can efficiently cleave aggrecan at several sites detected in joints of osteoarthritic patients. Although recent studies have shown that removal of the prodomain of ADAMTS4 is critical for its ability to degrade aggrecan, the cellular mechanisms for its processing and trafficking remain unclear. In this study, by using both furin-specific inhibitor and RNA interference technique, we demonstrate that furin plays an important role in the intracellular removal of ADAMTS4 prodomain. Further, we demonstrate that proADAMTS4 can be processed by means of multiple furin recognition sites: (206)RPRR(209), (209)RAKR(212), or (211)KR(212). The processing of proADAMTS4 was completely blocked by brefeldin A treatment, suggesting that processing occurs in the trans-Golgi network. Indeed, ADAMTS4 is co-localized with furin in trans Golgi network. Interestingly, the pro form of ADAMTS4, not its mature one, co precipitates with furin, suggesting that furin physically interacts with the prodomain of ADAMTS-4. In addition, our evidence suggests that a furin independent pathway may also contribute to the activation of ADAMTS4. These results indicate that the activation mechanism for ADAMTS4 can be targeted for therapeutical intervention against this enzyme. PMID- 14744862 TI - A monomer-to-trimer transition of the human mitochondrial transcription termination factor (mTERF) is associated with a loss of in vitro activity. AB - The human mitochondrial transcription termination factor (mTERF) is a nuclear encoded 39-kDa protein that recognizes a mtDNA segment within the mitochondrial tRNA(Leu(UUR)) gene immediately adjacent to and downstream of the 16 S rRNA gene. Binding of mTERF to this site promotes termination of rDNA transcription. Despite the fact that mTERF binds DNA as a monomer, the presence in its sequence of three leucine-zipper motifs suggested the possibility of mTERF establishing intermolecular interactions with proteins of the same or different type. When a mitochondrial lysate from HeLa cells was submitted to gel filtration chromatography, mTERF was eluted in two peaks, as detected by immunoblotting. The first peak, which varied in proportion between 30 and 50%, appeared at the position expected from the molecular mass of the monomer (41 +/- 2 kDa), and the gel filtration fractions that contained it exhibited DNA binding activity. Most interestingly, the material in this peak had a strong stimulating activity on in vitro transcription of the mitochondrial rDNA. The second peak eluted at a position corresponding to an estimated molecular mass of 111 +/- 5 kDa. No mTERF DNA binding activity could be detected in the corresponding gel filtration fractions. Therefore, we propose that mTERF exists in mitochondria in two forms, an active monomer and an inactive large size complex. The estimated molecular weight of this complex and the fact that purified mTERF can be eluted from a gel filtration column as a complex of the same molecular weight strongly suggest that this inactive complex is a homotrimer of mTERF. PMID- 14744863 TI - The aqueous accessibility in the external half of transmembrane domain I of the GABA transporter GAT-1 Is modulated by its ligands. AB - The sodium- and chloride-dependent gamma-aminobutyric acid (GABA) transporter GAT 1 is the first identified member of a family of transporters, which maintain low synaptic neurotransmitter levels and thereby enable efficient synaptic transmission. To obtain evidence for the idea that the highly conserved transmembrane domain I (TMD I) participates in the permeation pathway, we have determined the impact of impermeant methanethiosulfonate (MTS) reagents on cysteine residues engineered into this domain. As a background the essentially insensitive but fully active C74A mutant has been used. Transport activity of mutants with a cysteine introduced cytoplasmic to glycine 63 is largely unaffected and is resistant to the impermeant MTS reagents. Conversely, transport activity in mutants extracellular to glycine 63 is strongly impacted. Nevertheless, transport activity could be measured in all but three mutants: G65C, N66C, and R69C. In each of the six active cysteine mutants the activity is highly sensitive to the impermeant MTS reagents. This sensitivity is potentiated by sodium in L64C, F70C, and Y72C, but is protected in V67C and P71C. GABA protects in L64C, W68C, F70C, and P71C. The non-transportable GABA analogue SKF100330A also protects in L64C, W68C, and P71C as well as V67C, but strikingly potentiates inhibition in F70C. Although cysteine substitution in this region may have perturbed the native structure of GAT-1, our observations, taken together with the recently published accessibility study on the related serotonin transporter (Henry, L. K., Adkins, E. M., Han, Q., and Blakely, R. D. (2003) J. Biol. Chem. 278, 37052-37063), suggest that the extracellular part of TMD I is conformationally sensitive, lines the permeation pathway, and forms a more extended structure than expected from a membrane-embedded alpha-helix. PMID- 14744864 TI - SH2-containing inositol phosphatase 2 predominantly regulates Akt2, and not Akt1, phosphorylation at the plasma membrane in response to insulin in 3T3-L1 adipocytes. AB - SH2-containing inositol phosphatase 2 (SHIP2) is a physiologically important negative regulator of insulin signaling by hydrolyzing the phosphatidylinositol (PI) 3-kinase product PI 3,4,5-trisphosphate in the target tissues of insulin. Targeted disruption of the SHIP2 gene in mice resulted in increased insulin sensitivity without affecting biological systems other than insulin signaling. Therefore, we investigated the molecular mechanisms by which SHIP2 specifically regulates insulin-induced metabolic signaling in 3T3-L1 adipocytes. Insulin induced phosphorylation of Akt, one of the molecules downstream of PI3-kinase, was inhibited by expression of wild-type SHIP2, whereas it was increased by expression of 5'-phosphatase-defective (DeltaIP) SHIP2 in whole cell lysates. The regulatory effect of SHIP2 was mainly seen in the plasma membrane (PM) and low density microsomes but not in the cytosol. In this regard, following insulin stimulation, a proportion of Akt2, and not Akt1, appeared to redistribute from the cytosol to the PM. Thus, insulin-induced phosphorylation of Akt2 at the PM was predominantly regulated by SHIP2, whereas the phosphorylation of Akt1 was only minimally affected. Interestingly, insulin also elicited a subcellular redistribution of both wild-type and DeltaIP-SHIP2 from the cytosol to the PM. The degree of this redistribution was inhibited in part by pretreatment with PI3 kinase inhibitor. Although the expression of a constitutively active form of PI3 kinase myr-p110 also elicited a subcellular redistribution of SHIP2 to the PM, expression of SHIP2 appeared to affect the myr-p110-induced phosphorylation, and not the translocation, of Akt2. Furthermore, insulin-induced phosphorylation of Akt was effectively regulated by SHIP2 in embryonic fibroblasts derived from knockout mice lacking either insulin receptor substrate-1 or insulin receptor substrate-2. These results indicate that insulin specifically stimulates the redistribution of SHIP2 from the cytosol to the PM independent of 5'-phosphatase activity, thereby regulating the insulin-induced translocation and phosphorylation of Akt2 at the PM. PMID- 14744865 TI - Munc-18-1 inhibits phospholipase D activity by direct interaction in an epidermal growth factor-reversible manner. AB - Mammalian phospholipase D (PLD) has been reported to be a key enzyme for epidermal growth factor (EGF)-induced cellular signaling, however, the regulatory mechanism of PLD is still unclear. In this report, we found that Munc-18-1 is a potent negative regulator of PLD in the basal state and that its inhibition is abolished by EGF stimulation. We investigated PLD-binding proteins obtained from rat brain extract, and identified a 67-kDa protein as Munc-18-1 by peptide-mass finger-printing. The direct association between PLD and Munc-18-1 was confirmed by in vitro binding analysis using the purified proteins, and their binding sites were identified as the phox homology domain of PLD and multiple sites of Munc-18 1. PLD activity was potently inhibited by Munc-18-1 in vitro (IC50 = 2-5 nm), and the cotransfection of COS-7 cells with Munc-18-1 and PLD inhibited basal PLD activity in vivo. In the basal state, Munc-18-1 coprecipitated with PLD and colocalized with PLD2 at the plasma membrane of COS-7 cells. EGF treatment triggered the dissociation of Munc-18-1 from PLD when PLD was activated by EGF. The dissociation of the endogenous interaction between Munc-18-1 and PLD, and the activation of PLD by EGF were also observed in primary cultured chromaffin cells. These results suggest that Munc-18-1 is a potent negative regulator of basal PLD activity and that EGF stimulation abolishes this interaction. PMID- 14744866 TI - YY1AP, a novel co-activator of YY1. AB - With the aim of identifying potential cellular proteins that mediate the transcriptional regulation of YY1, a HeLa cDNA library was screened using the yeast two-hybrid system. A previously unknown protein interacting with YY1 was identified and named YY1AP. By using the 5'-rapid amplification of cDNA ends technique, the full-length cDNA of YY1AP was cloned and sequenced. The cDNA was 2253 bp in length and encoded an open reading frame of 750 amino acids. The chromosomal gene was made up of 10 exons separated by nine introns and is localized on chromosome 1 (1q21.3). Northern blot analysis revealed that YY1AP is ubiquitously expressed in various human tissues and cancer cell lines. Co immunoprecipitation and immunostaining of cells further indicated that YY1AP co localizes with YY1 in the nucleus. Furthermore, YY1AP was shown to be capable of enhancing the transcriptional activation of an YY1 responsive promoter. Subsequent analysis by glutathione S-transferase pull-down assay showed that YY1AP contained two YY1 binding regions. The transactivation region of YY1AP would seem to be localized within the section of amino acids 260-345. It is proposed that YY1AP is a novel co-activator of YY1. PMID- 14744867 TI - Cyclic AMP activation of the extracellular signal-regulated kinases 1 and 2: implications for intestinal cell survival through the transient inhibition of apoptosis. AB - The proliferative compartment of the intestinal crypt is critical in the process of intestinal epithelial cell homeostasis. The ability of these progenitor crypt cells to resist apoptosis and ensure restitution during a potentially lethal insult, but retain the ability to remove damaged or altered cells afterward, is necessary for preservation of the crypt-villus unit. We have examined the ability of cAMP to transiently inhibit apoptosis via the extracellular signal-regulated kinases 1 and 2 (ERK1/2), in T84 cells, an intestinal crypt-like cell line. Using the cAMP analog 8-bromo-cAMP and cholera toxin (CT), cAMP-mediated ERK1/2 activation was first measured by Western blot analysis of the phosphorylated (activated) and total (activated and inactivated) forms of ERK1/2. Cyclic AMP activated ERK1/2 in a time- and dose-dependent manner, and the effect was inhibited by PD098059, an inhibitor of the ERK1/2 signaling pathway. However, inhibition of protein kinase A (PKA) did not alter the activation of ERK1/2. CT transiently inhibited both staurosporine and Fas antibody mediated apoptosis as measured by a caspase-3 activation assay and the detection of nucleosomes in an apoptosis based enzyme-linked immunosorbent assay. This inhibitory effect was reversed by the simultaneous addition of PD098059. Our data suggest that in the T84 cell line, cAMP activates ERK1/2 in a PKA independent fashion and a physiological consequence of this activated pathway is the transient inhibition of apoptosis. These findings suggest a novel pathway that intestinal cells use to protect against injury while maintaining the overall ability to remove damaged cells and preserve intestinal homeostasis. PMID- 14744868 TI - Identification of novel binding elements and gene targets for the homeodomain protein BARX2. AB - BARX2 is a homeobox transcription factor that influences cellular differentiation in various developmental contexts. To begin to identify the gene targets that mediate its effects, chromatin immunoprecipitation (ChIP) was used to isolate BARX2 binding sites from the human MCF7 breast cancer cell line. Cloning and sequencing of BARX2-ChIP-derived DNA fragments identified 60 potential BARX2 target loci that were proximal to or within introns of genes involved in cytoskeletal organization, cell adhesion, growth factor signaling, transcriptional regulation, and RNA metabolism. The sequences of over half of the fragments showed homology with the mouse genome, and several sequences could be mapped to orthologous human and mouse genes. Binding of BARX2 to 21 genomic loci examined was confirmed quantitatively by replicate ChIP assays. A combination of sequence analysis and electrophoretic mobility shift assays revealed homeodomain binding sites within several fragments that bind to BARX2 in vitro. The majority of BARX2 binding fragments tested (14/19), also affected transcription in luciferase reporter gene assays. Mutation analyses of three fragments showed that their transcriptional activities required the HBS, and suggested that BARX2 regulates gene expression by binding to DNA elements containing paired TAAT motifs that are separated by a poly(T) sequence. Inhibition of BARX2 expression in MCF7 cells led to reduced expression of eight genes associated with BARX2 binding sites, indicating that BARX2 directly regulates their expression. The data suggest that BARX2 can coordinate the expression of a network of genes that influence the growth of MCF7 cells. PMID- 14744869 TI - SREBP-1c and Sp1 interact to regulate transcription of the gene for phosphoenolpyruvate carboxykinase (GTP) in the liver. AB - The sterol regulatory element-binding protein-1c (SREBP-1c), as well as SREBP-1a and SREBP-2, inhibit transcription of the gene encoding the cytosolic form of phosphoenolpyruvate carboxykinase (GTP) (PEPCK-C). There are two SREBP regulatory elements (SREs) in the PEPCK-C gene promoter (-322 to -313 and -590 to -581). The SRE at -590 overlaps an Sp1 site on the opposite strand of the DNA. These SREs bound SREBP-1a and SREBP-1c with low affinity but the addition of purified upstream stimulatory activity enhanced the binding of SREBP-1 to both of these sites. Mutating these SREs increased both unstimulated (5-fold) and protein kinase A-stimulated transcription (8-27-fold) from the PEPCK-C gene promoter; this was lost when both SREs were mutated. The SRE at -590 differs by a single base pair from the SRE in the low density lipoprotein (LDL) receptor gene (T in the PEPCK-C gene promoter at -582, compared with an A in the SRE of the gene for the LDL receptor promoter). Introduction of the LDL receptor SRE into the PEPCK-C gene promoter increased SREBP-1c binding and caused a 10-fold enhancement of basal transcription from the promoter, rather than an inhibition as observed with the SRE in the PEPCK-C gene promoter. The T/A change does not alter the binding of Sp1 to its site on the opposite strand of the DNA. Sp1 bound to the promoter independently of SREBP-1c but competed with SREBP-1c for binding. Sp1 does not bind to the SRE at -322. Chromatin immunoprecipitation analysis, using rat hepatocytes, demonstrated that SREBP-1 and Sp1 were associated in vivo with putative regulatory regions corresponding to the SREs in the PEPCK-C gene promoter. We propose that insulin represses transcription of the gene for PEPCK-C by inducing SREBP-1c production in the liver, which interferes with the stimulatory effect of Sp1 at -590 of the PEPCK-C gene promoter. PMID- 14744870 TI - Progesterone receptor deficient in chromatin binding has an altered cellular state. AB - Our previous work has shown that the progesterone receptor (PR) can exist in two distinct functional states in mammary adenocarcinoma cells. The differences in function included the ability to activate a promoter in organized chromatin, sensitivity to ligand, and ligand-independent activation. To determine whether these functional differences were because of altered cellular processing, we carried out biochemical analyses of the functionally distinct PRs. Although the majority of PR is localized to the nucleus, biochemical partitioning resulted in a loosely bound (cytosolic) fraction, and a tightly bound (nuclear) fraction. In the absence of progestins, the functionally distinct PRs differed significantly in partitioning between the two fractions. To characterize these fractions further, we analyzed interactions of unliganded PR with chaperones by coimmunoprecipitation. We determined that PR in the cytosolic fraction associated with hsp90 and p23. In contrast, PR in the nuclear fraction consisted of complexes containing hsp90, p23, and FKBP51 as well as PR that was dimerized and highly phosphorylated. Hormone treatment significantly reduced the formation of all PR-chaperone complexes. The hsp90 inhibitor, geldanamycin, similarly blocked transcriptional activity of both functionally distinct receptors. However, the two forms of the PR differed in their ability to associate with the mouse mammary tumor virus promoter in organized chromatin. These findings provide new information about the composition and distribution of mature progesterone receptor complexes in mammary adenocarcinoma cells, and suggest that differences in receptor subcellular distribution have a significant impact on their function. These findings also reveal that transiently expressed steroid receptors may not always be processed like their endogenous counterparts. PMID- 14744871 TI - Degradation of mutated bovine pancreatic trypsin inhibitor in the yeast vacuole suggests post-endoplasmic reticulum protein quality control. AB - The rate-limiting step in protein secretion is folding, which occurs in the endoplasmic reticulum (ER) lumen, and almost all secreted proteins contain disulfide bonds that form in the ER and stabilize the native state. Secreted proteins unable to fold may aggregate or they may be subject to ER-associated protein degradation. To examine the fate of aberrant forms of a well characterized, disulfide-bonded secreted protein, we expressed bovine pancreatic trypsin inhibitor in yeast. Bovine pancreatic trypsin inhibitor is a single domain, 58-amino acid polypeptide containing three disulfide bonds, and yeast cells secrete the wild type protein. In contrast, the Y35L mutant, which folds rapidly but is unstable, remains soluble and is not secreted. Surprisingly, the proteolysis of Y35L is unaffected in yeast containing mutations in genes encoding factors required for ER-associated protein degradation and is stable if artificially retained in the ER. Rather, Y35L is diverted from the Golgi to the vacuole and degraded. Because only the mutant protein is quantitatively proteolyzed these data suggest that a post-ER quality control check-point diverts unstable proteins to the vacuole for degradation. PMID- 14744872 TI - Identification of a role for beta-catenin in the establishment of a bipolar mitotic spindle. AB - beta-Catenin is a multifunctional protein that is known to participate in two well defined cellular processes, cell-cell adhesion and Wnt-stimulated transcriptional activation. Here we report that beta-catenin participates in a third cellular process, the establishment of a bipolar mitotic spindle. During mitosis, beta-catenin relocalizes to mitotic spindle poles and to the midbody. Furthermore, biochemical fractionation demonstrates the presence of beta-catenin in purified centrosome preparations. Reduction of cellular beta-catenin by RNA interference leads to the failure of centrosomes to fully separate, resulting in a marked increase in the frequency of monoastral mitotic spindles. Our results define a new and important function for beta-catenin in mitosis and demonstrate that beta-catenin is involved in vital biological processes beyond cell adhesion and Wnt signaling. PMID- 14744873 TI - Immunohistochemical classification and functional morphology of human choroidal ganglion cells. AB - PURPOSE: To characterize human choroidal ganglion cells (CGCs) further, regarding their immunohistochemical and ultrastructural appearance and their pre- and postsynaptic connections. METHODS: . Choroidal wholemounts and serial sections of human donor eyes were stained with antibodies against neuronal nitric oxide synthase (nNOS), vasoactive intestinal peptide (VIP), tyrosine hydroxylase (TH), vesicular monoaminergic transporter (VMAT)-2, vesicular acetylcholine transporter (VAChT), neuropeptide Y (NPY), substance P (SP), calcitonin gene-related peptide (CGRP), calretinin, galanin, synaptophysin, and alpha-smooth muscle actin. Ultrathin sections of glutaraldehyde-fixed eyes were studied with an electron microscope. RESULTS: All CGCs stained for nNOS, most for VIP, approximately 45% for calretinin, and only single neurons for NPY and galanin. Ultrastructurally, the CGCs showed an incomplete glial sheath and, in places, showed close contact to surrounding collagen fibrils. The CGCs were in close contact with numerous boutons staining for the different neuronal markers including synaptophysin, nNOS, VIP, NPY, TH, VMAT-2, VAChT, calretinin, and NPY. CONCLUSIONS: The data indicate a complex integrative function of CGCs. The immunohistochemical and ultrastructural characteristics also indicate that the CGCs may have mechanosensory properties. The complex synaptic information points to a specific regulative CGC function in parallel with ciliary muscle contraction (accommodation). Axons originating from CGCs mainly supply the choroidal vasculature, thus implicating the CGCs as vasodilative neurons, but single CGCs may also innervate other structures such as nonvascular choroidal smooth muscle cells. PMID- 14744874 TI - VEGF164(165) as the pathological isoform: differential leukocyte and endothelial responses through VEGFR1 and VEGFR2. AB - PURPOSE: Vascular endothelial growth factor (VEGF) induces angiogenesis and vascular permeability and is thought to be operative in several ocular vascular diseases. The VEGF isoforms are highly conserved among species; however, little is known about their differential biological functions in adult tissue. In the current study, the inflammatory potential of two prevalent VEGF isoform splice variants, VEGF(120(121)) and VEGF(164(165)), was studied in the transparent and avascular adult mouse cornea. METHODS: Controlled-release pellets containing equimolar amounts of VEGF(120) and VEGF(164) were implanted in corneas. The mechanisms underlying this differential response of VEGF isoforms were explored. The response of VEGF in cultured endothelial cells was determined by Western blot analysis. The response of VEGF isoforms in leukocytes was also investigated. RESULTS: VEGF(164) was found to be significantly more potent at inducing inflammation. In vivo blockade of VEGF receptor (VEGFR)-1 significantly suppressed VEGF(164)-induced corneal inflammation. In vitro, VEGF(165) more potently stimulated intracellular adhesion molecule (ICAM)-1 expression on endothelial cells, an effect that was mediated by VEGFR2. VEGF(164) was also more potent at inducing the chemotaxis of monocytes, an effect that was mediated by VEGFR1. In an immortalized human leukocyte cell line, VEGF(165) was found to induce tyrosine phosphorylation of VEGFR1 more efficiently. CONCLUSIONS: Taken together, these data identify VEGF(164(165)) as a proinflammatory isoform and identify multiple mechanisms underlying its proinflammatory biology. PMID- 14744875 TI - Delayed expression of the Crx gene and photoreceptor development in the Chx10 deficient retina. AB - PURPOSE: The Chx10 homeobox gene is expressed in neural progenitor cells during retinal development. The absence of Chx10 causes microphthalmia in humans and in the mouse mutant ocular retardation. The purpose of this study was to examine how neuronal development is affected by absence of the Chx10 transcription factor in the mouse retina. METHODS: Expression of transcription factor genes, Crx, Pou4f2, and Pax6, that mark specific cell types as they begin to differentiate was analyzed by RNA in situ hybridization of retina from wild-type and Chx10-null ocular retardation mice (Chx10(or-J/or-J)). RT-PCR analysis was used to compare expression of these genes and putative targets of Crx regulation. Photoreceptor development was analyzed by using peanut agglutinin (PNA)-rhodamine and blue cone opsin antibody to label cones and rhodopsin antibody to label rods. RESULTS: The photoreceptor gene Crx, normally expressed during embryonic retinal development, was not detected in the embryonic mutant retina, but was expressed after birth. Expression of the targets of Crx regulation, rhodopsin, peripherin, rod phosphodiesterase beta (Pdeb), and arrestin, with the exception of interphotoreceptor retinoid binding protein (Irbp), was delayed in the Chx10(or J/or-J) retina. Rhodopsin localization in rod outer segments was also delayed. By contrast, temporal and spatial expression of Pou4f2 and Pax6 in developing ganglion and amacrine cells and PNA and blue opsin in developing cone cells was relatively normal in the mutant. CONCLUSIONS: Delay of the normal temporal expression of genes essential for photoreceptor disc morphogenesis leads to failure of correct rod and cone outer segment formation in the Chx10(or-J/or-J) mutant retina. In addition, the absence of Chx10 appears to affect the development of late-born cells more than that of early-born cells, in that a low number of rods develops, whereas formation of ganglion, amacrine, and cone cells is relatively unaffected. PMID- 14744876 TI - Functional properties of natural human PAX6 and PAX6(5a) mutants. AB - PURPOSE: Pax6 is essential for development of the eye, brain, and pancreas. Two major products of PAX6 are specific DNA-binding proteins, PAX6 and PAX6(5a). PAX6(5a) contains a short insertion influencing its DNA-binding activity. Heterozygous mutations in PAX6 result in abnormal eye development implicating haploinsufficiency. Deletions of one PAX6 allele result in aniridia characterized by severe ocular phenotypes. Approximately 10% of PAX6 mutations encode missense mutations. These mutations usually cause less severe abnormalities than does aniridia. The moderate phenotypes raise the possibility that different ocular tissues are differently sensitive to specific mutations. To test this hypothesis, we probed functional properties of individual mutated Pax6 proteins in a variety of conditions. METHODS: Mutations in PAX6 and PAX6(5a) were introduced by site directed mutagenesis and tested by transfections in four cell lines using reporters containing three different Pax6 binding sites. Pax6 binding to DNA was studied by electrophoretic mobility shift assays. RESULTS: Functional studies of PAX6 and PAX6(5a) and their eight natural missense (G18W, R26G, A33P, S43P, G64V, I87R, V126D and R128C) and two nonsense (R317X and S353X) disease-causing mutants revealed unexpected pleiotropic effects in gene regulation, not predicted by the PAX6-DNA crystal structure. Transactivation by PAX6 and PAX6(5a) was dependent on the location of mutation, type of DNA-binding site, and cellular environment. CONCLUSIONS: This work provides evidence that activation by PAX6 and PAX6(5a) is modulated by specific cellular environments. It is likely that moderate phenotypes associated with PAX6 missense mutations originate from abnormal protein function in a restricted number of ocular cell types. PMID- 14744877 TI - Cellular retinaldehyde-binding protein interacts with ERM-binding phosphoprotein 50 in retinal pigment epithelium. AB - PURPOSE: To characterize mechanisms of apical localization of visual cycle components in retinal pigment epithelium (RPE) by the identification of cellular retinaldehyde-binding protein (CRALBP) interaction partners. METHODS: An overlay assay was used to detect interactions of CRALBP with components of RPE microsomes. Interacting proteins were identified with two-dimensional (2D)-PAGE and liquid chromatography tandem mass spectrometry (LC MS/MS). Protein interactions were characterized by affinity chromatography, peptide competition, and expression of protein domains. Protein colocalization in mouse retina was examined using double-label immunocytochemistry and confocal microscopy. RESULTS: CRALBP bound to a 54-kDa protein in RPE microsomes, which was identified as ERM (ezrin, radixin, moesin)-binding phosphoprotein 50 (EBP50), a PDZ domain protein, also known as sodium/hydrogen exchanger regulatory factory type 1 (NHERF-1). EBP50 and ezrin in solubilized microsomes bound to CRALBP-agarose but not to a control agarose column. CRALBP bound to both recombinant PDZ domains of EBP50 but not to the C-terminal ezrin-binding domain. In outer retina, EBP50 and ezrin were localized to RPE and Muller apical processes. CRALBP was distributed throughout both RPE and Muller cells, including their apical processes. CONCLUSION: RM proteins are multivalent linkers that connect plasma membrane proteins with the cortical actin cytoskeleton. EBP50 interacts with ERM family members through a C terminal domain and binds targets such as CRALBP through its PDZ domains, thus contributing to an apical localization of target proteins. Our results provide a structural basis for apical localization of a retinoid-processing complex in RPE cells and offer insight into the cell biology of retinoid processing and trafficking in RPE. PMID- 14744878 TI - Effects of positive and negative lens treatment on retinal and choroidal glucagon and glucagon receptor mRNA levels in the chicken. AB - PURPOSE: It has been found in the chicken that the amount of retinal glucagon mRNA increases during treatment with positive lenses. Pharmacological studies support the idea that glucagon may act as a stop signal for visually induced eye growth. To gain more insight into the functional role of glucagon, the changes of glucagon and glucagon receptor mRNA concentrations in retina and choroid over time were studied. Furthermore, the abundance of glucagon and the glucagon receptor was studied in different fundal layers (retina, retinal pigment epithelium[RPE], choroid) and the blood. METHODS: Semiquantitative real-time RT PCR was used to measure glucagon and glucagon receptor mRNA levels in retina and choroid after positive and negative lens treatment for 2, 6, or 24 hours, by unilateral -7- or +7-D lenses. Contralateral eyes served as the control, and completely untreated animals provided further reference data. Intravitreal colchicine injections (which are known to reduce the number of glucagon cells sharply) were used to verify that the related decline in glucagon mRNA could be measured by real-time RT-PCR. RESULTS: In the retina, treatment with -7-D lenses induced an initial upregulation of glucagon mRNA in both eyes, followed by a significant downregulation. The treatment with +7-D lenses showed a significant but transient downregulation in the control eye superimposed on a trend toward upregulation in the treated eye. However, the changes in glucagon mRNA expression were not confined to the lens-treated eyes but were also found, although sometimes to a lesser extent, in the non-lens-covered fellow eyes. There was evidence of a transient increase in glucagon receptor mRNA levels in lens-treated eyes after either -7- or +7-D lens wear. In the choroid, no effect of imposed defocus was detected. The injection of colchicine led to the destruction of approximately 75% of the glucagon amacrine cells but the mRNA level of retinal glucagon decreased by only approximately 50%. Glucagon receptor expression was found to be higher in the RPE than the retina and choroid whereas, in the blood, glucagon and glucagon receptor mRNA expression was below detection level. CONCLUSIONS: The observed bidirectional regulation of glucagon mRNA in correlation with the sign of imposed defocus supports the idea that glucagon may act as a stop-and-go signal for eye growth. This is in line with a previous proposal based on studies of changes of the glucagon peptide content. PMID- 14744879 TI - Correction of refractive error in the adult population of Bangladesh: meeting the unmet need. AB - PURPOSE: To assess the extent of uncorrected refractive error and associated factors in Bangladesh and to suggest ways in which this need can be met. METHODS: A nationally representative sample of 12,782 adults (>/= 30 years of age) was selected. Of them, 11,624 subjects underwent a demographic interview, visual acuity (logarithm of the minimum angle of resolution [logMAR]) measurement, automated refraction, and optic disc examination. Subjects with visual acuity less than 6/12 in either eye also had a corrected refraction measurement, cataract grading, and dilated retinal examination. RESULTS: Of the 11,624 subjects examined, 2,469 (22.1%) were myopes (less than -0.5 D) and 2,308 (20.6%) hyperopes (more than +0.5 D). The spectacle coverage percentage, calculated as [met need/(met need + unmet need) x 100%] was 25.2% and 40.5%, using 6/12 and 6/18 visual acuity cutoffs, respectively, and was higher in men and urban inhabitants. Older subjects and the literate and more highly educated were more likely to wear spectacles; however, most spectacle wearers (81%) had inadequate correction. Of the 1142 subjects who would benefit from spectacles, 827 (72.4%) would be suitable for off-the-shelf spectacles. Subjects without spectacles with less than 6/12 in the better eye (n = 835), would achieve 6/12 or better with correction (unmet need). Extrapolation to the national population yields an estimate that 1.5 million (6.7%) adult men and 1.8 million (9.2%) women have an unmet need for refractive correction. CONCLUSIONS: In Bangladesh, there is low spectacle coverage with a large unmet need. This survey identified risk groups, in particular women and those living in rural areas. This description of the availability of refractive services suggests areas for improvement (e.g., off-the shelf spectacles) that may enable Bangladesh to achieve the goals of the World Health Organization's Vision 2020 initiative. PMID- 14744880 TI - Corneal nerve tortuosity in diabetic patients with neuropathy. AB - PURPOSE: Corneal confocal microscopy is a reiterative, rapid, noninvasive in vivo clinical examination technique capable of imaging corneal nerve fibers. Nerve fiber tortuosity may indicate a degenerative and attempted regenerative response of nerve fibers to diabetes. The purpose of this study was to define alterations in the tortuosity of corneal nerve fibers in relation to age, duration of diabetes, glycemic control, and neuropathic severity. METHODS: The cornea and collected images of the subbasal nerve plexus of 18 diabetic patients (stratified into mild, moderate, and severe neuropathic groups using conventional clinical measures of neuropathy) and 18 age-matched nondiabetic control subjects were scanned, and a novel mathematical paradigm was applied to quantify the extent of nerve tortuosity, which was termed the tortuosity coefficient (TC). RESULTS: TC was significantly different between the four clinical groups (F(3) = 12.2, P < 0.001). It was significantly greater in the severe neuropathic group than in control subjects (P < 0.003) and in the mild (P < 0.004) and moderate (P < 0.01) neuropathic groups. TC did not correlate significantly with the age (r = -0.003, P > 0.05), duration of diabetes (r = -0.219, P > 0.05), or hemoglobin A1c (HbA1c; r = 0.155, P > 0.05) of diabetic patients. CONCLUSIONS: Corneal confocal microscopy allows rapid, noninvasive in vivo evaluation of corneal nerve tortuosity. This morphologic abnormality relates to the severity of somatic neuropathy and may reflect an alteration in the degree of degeneration and regeneration in diabetes. PMID- 14744881 TI - Morphological and functional changes in the rat cornea with an ethanol-mediated epithelial flap. AB - PURPOSE: To establish morphologic and functional changes in the rat cornea after 20% ethanol-mediated epithelial flap creation. METHODS: The epithelial flap was detached using 20% ethanol and then repositioned. On the other eye, the corneal epithelium was mechanically scraped. The morphologic changes were examined by light and transmission electron microscopy (LM and TEM). Terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) assays and proliferative cell nuclear antigen (PCNA) staining were performed to detect apoptosis and proliferation, respectively. The number of central stromal keratocytes was counted during wound-healing periods. A Y-chromosome-specific gene (SRY) was detected in genomic DNA obtained from female epithelial cells with male epithelial flap. Expressions of matrix metalloproteinase (MMP)-9, aquaporine5 (AQP5), and mucin1 (MUC1) mRNAs were examined by reverse transcription-polymerase chain reaction. RESULTS: After flap repositioning, damaged basal cells were observed until day 1, and the death of epithelial cells and stromal keratocytes loss peaked at day 1. Cell proliferation and MMP-9 mRNA levels peaked from days 1 to 3. By contrast, after mechanical scraping, the denuded stromal surface was covered with multilayer epithelial cells at day 3, cell death peaked at 4 hours, cell proliferation peaked from 12 hours to day 1, stromal keratocytes loss peaked at 8 hours, and MMP-9 mRNA was widely expressed from 12 hours to day 3. In both corneas, hemidesmosomes were not observed until day 1, and they continued to be present at day 3. The SRY gene was detected in female epithelial cells at day 1 after transplantation, but not at day 3. Expressions of AQP5 and MUC1 mRNAs were unchanged. CONCLUSIONS: Ethanol-mediated flap creation induces less keratocyte loss and a slower wound-healing process than mechanical scraping. The flap protecting the underlying stromal surface may have epithelial function. In addition, this Sprague-Dawley rat model may be useful in the study of the wound-healing response after LASEK. PMID- 14744882 TI - Postnatal gene expression in the normal mouse cornea by SAGE. AB - PURPOSE: To provide a detailed gene expression profile of the normal postnatal mouse cornea. METHODS: Serial analysis of gene expression (SAGE) was performed on postnatal day (PN)9 and adult mouse (6 week) total corneas. The expression of selected genes was analyzed by in situ hybridization. RESULTS: A total of 64,272 PN9 and 62,206 adult tags were sequenced. Mouse corneal transcriptomes are composed of at least 19,544 and 18,509 unique mRNAs, respectively. One third of the unique tags were expressed at both stages, whereas a third was identified exclusively in PN9 or adult corneas. Three hundred thirty-four PN9 and 339 adult tags were enriched more than fivefold over other published nonocular libraries. Abundant transcripts were associated with metabolic functions, redox activities, and barrier integrity. Three members of the Ly-6/uPAR family whose functions are unknown in the cornea constitute more than 1% of the total mRNA. Aquaporin 5, epithelial membrane protein and glutathione-S-transferase (GST) omega-1, and GST alpha-4 mRNAs were preferentially expressed in distinct corneal epithelial layers, providing new markers for stratification. More than 200 tags were differentially expressed, of which 25 mediate transcription. CONCLUSIONS: In addition to providing a detailed profile of expressed genes in the PN9 and mature mouse cornea, the present SAGE data demonstrate dynamic changes in gene expression after eye opening and provide new probes for exploring corneal epithelial cell stratification, development, and function and for exploring the intricate relationship between programmed and environmentally induced gene expression in the cornea. PMID- 14744883 TI - Mouse strain-dependent heterogeneity of resting limbal vasculature. AB - PURPOSE: Heterogeneity of the extent of angiogenesis induced by exogenous growth factors may be determined by genetic influences. Because angiogenesis is the formation of new vessels from preexisting ones, strain-related influences on naive resting limbal vessel phenotype and gene expression were determined in mice having divergently low and high angiogenic responses. METHODS: Resting limbal vessel surface area and density and extent of bFGF-induced corneal angiogenesis were determined in C57BL/6J, BALB/cJ, F1 intercross identical with C57BL/6J X 129S3/SvIM, and 129S3/SvIM mouse strains by quantitative three-dimensional reconstruction confocal microscopy. Strain-related influences on pro- and antiangiogenic gene expression in naive cornea were determined by quantitative real-time RT-PCR. RESULTS: The strain-dependent rank order of resting limbal vessel surface area and resting vessel density paralleled bFGF-induced neovascularization: 129S3/SvIM > BALB/cJ, F1 > C57BL/6J (P < 0.0006). Pigment epithelium-derived factor (PEDF) was increased more than 67-fold compared to Ang 2 in resting cornea of both C57BL/6J and 129S3/SvIM strains (P < 0.0001; P < 0.0001), suggesting a strongly antiangiogenic environment. The corneas of the C57BL/6J mice demonstrated 1.8-, 1.5-, and 1.7-fold increased mRNA levels for Flt 1, VEGF, and bFGF, respectively (P < 0.02; P < 0.04; P < 0.02); however, TSP-1 expression was increased 2.4-fold compared with 129S3/SvIM (P < 0.0004). CONCLUSIONS: Strain-dependent differences in the resting limbal vessel surface area and density correlated with heterogeneity in the extent of bFGF-induced angiogenesis. Differences in pro- and antiangiogenic gene expression levels in resting cornea may influence vascular limbal phenotype during quiescence and may predict susceptibility to angiogenesis-dependent diseases. PMID- 14744884 TI - Promotion of corneal allograft survival by the induction of oxidative macrophages. AB - PURPOSE: A Th1-type immune response was detected in allotransplanted, rejected corneas. Because the intracellular thiol redox status of antigen-presenting cells (APCs) reportedly regulates the Th1/Th2 balance through distinctive cytokine production by APCs, this study was conducted to investigate the effect of the intracellular thiol redox status of macrophages (Mps) on corneal allograft survival. METHODS: N,N'-diacetyl-L-cystine dimethylester (NACOMe)(2) was injected intraperitoneally into BALB/c (H-2(d)) mice to induce Mps with a low intracellular glutathione content (icGSH). Corneal grafts from C57BL/10 (H-2(b)), B10.D2 (H-2(d)), and DBA/2 (H-2(d)) donor mice were placed on neovascularized BALB/c graft beds for assessment. B10.D2-grafted recipients were evaluated for donor-specific delayed-type hypersensitivity (DTH), and the cytokines produced by their lymphocytes were examined (IFN-gamma, IL-4, and IL-10). In other experiments, naive BALB/c mice, injected intravenously with Mps of low icGSH content, received B10.D2 corneal grafts. RESULTS: In (NACOMe)(2)-treated mice, 13 of 20 B10.D2 grafts and 6 of 10 DBA/2 grafts survived indefinitely. No grafts survived in the control mice (P < 0.0001). (NACOMe)(2) treatment did not enhance C57BL/10 graft survival. At 2 weeks after B10.D2 grafting, control mice exhibited DTH, but (NACOMe)(2)-treated mice did not (P < 0.01). Lymphocytes from (NACOMe)(2)-treated mice did not respond to donor splenocytes. Those of control mice showed Th1-type cytokine secretion. The intravenous transfer of peritoneal Mps from (NACOMe)(2)-treated mice prolonged corneal allograft survival (P < 0.003). CONCLUSIONS: The observed enhanced graft acceptance may be due to the suppression of alloantigen-induced Th1 polarization through the induction of Mps with reduced icGSH levels. PMID- 14744885 TI - Ocular rotation axes during dynamic Bielschowsky head-tilt testing in unilateral trochlear nerve palsy. AB - PURPOSE: To explain the positive Bielschowsky head-tilt (BHT) sign in unilateral trochlear nerve palsy (uTNP) by the kinematics of three-dimensional eye rotations. METHODS: Twelve patients with uTNP monocularly fixed on targets on a Hess screen were oscillated (+/- 35 degrees, 0.3 Hz) about the roll axis on a motorized turntable (dynamic BHT). Three-dimensional eye movements were recorded with dual search coils. Normal data were collected from 11 healthy subjects. RESULTS: The rotation axis of the viewing paretic or unaffected eye was nearly parallel to the line of sight. The rotation axis of the covered fellow eye, however, was tilted inward relative to the other axis. This convergence of axes increased with gaze toward the unaffected side. Over entire cycles of head roll, the rotation axis of either eye remained relatively stable in both the viewing and covered conditions. CONCLUSIONS: In patients with uTNP, circular gaze trajectories of the covered paretic or unaffected eye during dynamic BHT are a direct consequence of the nasal deviation of the rotation axis from the line of sight. This, in turn, is a geometrical result of decreased force by the superior oblique muscle (SO) of the covered paretic eye or, according to Hering's law, increased force parallel to the paretic SO in the covered unaffected eye. The horizontal incomitance of rotation axes along horizontal eye positions can be explained by the same mechanism. PMID- 14744886 TI - Perimetric defects and ganglion cell damage: interpreting linear relations using a two-stage neural model. AB - PURPOSE: To better understand the relations between glaucomatous perimetric defects and ganglion cell damage, a neural model was developed to interpret empiric findings on linear relations between perimetric defects and measures of ganglion cell loss. METHODS: A two-stage model computed responses of ganglion cell mosaics (first stage), then computed perimetric sensitivity in terms of processing by spatial filters (second stage) that pool the ganglion cell responses. Cell death and dysfunction were introduced in a local patch of the first-stage ganglion cell mosaic, and perimetric defect depth was computed for the corresponding region of the visual field. Calculations were performed for both sparse and dense ganglion cell mosaics and for spatial filters with peak frequencies from 0.5 to 4.0 cyc/deg. RESULTS: The model yielded nonlinear functions for perimetric defect depth in decibel versus the percentage of ganglion cell damage, but functions for lower spatial frequencies became linear when perimetric defect was expressed as a percentage of normal. The relations between perimetric defects and percentage of ganglion cell loss were determined primarily by spatial tuning of the second-stage spatial filters. For averaging sensitivities across different visual field locations, linear units (arithmetic mean) can more closely approximate mean ganglion cell loss than decibel units (geometric mean). Fits to data from experimental glaucoma required ganglion cell dysfunction in addition to ganglion cell loss. CONCLUSIONS: Pooling by second stage spatial filters can account for empiric findings of linear relations between perimetric defects and measures of ganglion cell loss. PMID- 14744887 TI - Selenium's effects on MMP-2 and TIMP-1 secretion by human trabecular meshwork cells. AB - PURPOSE: Because of the observed increase in incidence of glaucoma among some individuals taking selenium as a dietary supplement, the present study was undertaken to investigate mechanisms of selenium-induced changes in homeostasis of human trabecular meshwork (HTM) cells. Specifically, the impact of selenium on matrix metalloproteinases (MMPs), their inhibitors (tissue inhibitors of metalloproteinases; TIMPs), and the second messengers that regulate MMP expression was investigated in an HTM cell culture model. METHODS: HTM cell cultures were treated with an organic selenium compound (methyl seleninic acid), and changes in secretion and activity of MMPs and TIMPs were analyzed by Western blot and zymography. Changes in extracellular-signal-related kinases 1 and 2 (ERK1/2) and phospho-ERK1/2 levels were monitored by Western blot analysis of whole-cell lysates prepared from selenium-treated cells. Photographs of cultures over time were used to document selenium-induced changes in cell morphology. RESULTS: Treatment of HTM cells with selenium for 24 hours at doses ranging from 1 to 10 micro M caused a dose-dependent decrease in the secretion of MMP-2 and TIMP-1. Treatment for 6 hours revealed a significant decrease in MMP-2 and TIMP-1 at the highest dose. MMP-1, -3, and -9 and TIMP-2 were either not detected or their secretion was not consistently influenced by selenium treatment. Selenium treatment caused a significant decrease in ERK1/2 phosphorylation, but no change in overall ERK protein levels. Selenium treatment resulted in dose-dependent, reversible changes in HTM cell-matrix associations. CONCLUSIONS: Selenium-induced changes in MMP-2/TIMP-1 secretion may alter the balance of extracellular matrix turnover in the conventional outflow pathway and cause an increase in intraocular pressure that eventually leads to glaucoma. PMID- 14744888 TI - Influence of molecular weight on intracameral dextran movement to the posterior segment of the mouse eye. AB - PURPOSE: Uveoscleral outflow provides a potential pathway to the posterior segment for drug delivery. In this study, the influence of molecular weight on the intraocular distribution of dextran molecules after intracameral injection in the mouse eye was investigated. METHODS: The anterior chambers of the eyes of 64 anesthetized NIH Swiss mice were perfused with various fluorescent dextran solutions (10, 40, 70, and 500 kDa) at 500 nL/min for 10 minutes. At 10, 20, or 60 minutes after the initiation of the anterior chamber perfusion, the mice were killed and tissue obtained for evaluation by fluorescence microscopy. RESULTS: Each of the different molecular weight dextrans were visible in the anterior chamber of the mouse eye after the termination of the experiments. The 10-kDa dextran was observed in the supraciliary space and the supraciliary sclera after 10 minutes and in the anterior sclera after 60 minutes of perfusion. The 40-kDa dextran was detected in the supraciliary space and the anterior sclera after 10 minutes and in the anterior choroid and sclera after 20 and 60 minutes, but not in the posterior segment. The 70-kDa dextran was observed in the supraciliary space and anterior choroid after 10 minutes. After 20 minutes, it was visible in the equatorial choroid. After 60 minutes, it was observed in the posterior choroid. The 500-kDa dextran was observed in the supraciliary space and the anterior choroid after 10 minutes and in the supraciliary sclera at 20 minutes. At 60 minutes, 500-kDa dextran was observed in the equatorial choroid, but not farther toward the posterior. CONCLUSIONS: The influence of molecular weight on the redistribution of macromolecules from the anterior chamber to the posterior globe in the mouse eye appears to be similar to primate eyes. These similarities include passage of all size dextrans through the proximal uveoscleral pathway, the dependence of the extent of posterior movement on the size of the dextran, and the absence of large dextran entry into the distal uveoscleral pathway. PMID- 14744889 TI - Expression of connective tissue growth factor after glaucoma filtration surgery in a rabbit model. AB - PURPOSE: Connective tissue growth factor (CTGF) appears to play a significant role in mediating fibrosis in several tissues. To gain further understanding of the role of CTGF in the scar formation that occurs after glaucoma filtering surgery (GFS), experiments were performed in a rabbit model. METHODS: . Three experiments were performed: (1) CTGF and transforming growth factor (TGF)-beta expression were measured quantitatively after GFS, using ELISA. (2) After GFS conjunctival bleb tissues were immunostained for the presence of CTGF and TGF beta. (3) Exogenous CTGF was injected into mitomycin-C (MMC)-treated filtering blebs and the scaring response compared to TGF-beta and physiological saline injected blebs. RESULTS: CTGF and TGF-beta were expressed maximally by day 5 after surgery and were both shown to be present in the bleb tissues after GFS. The addition of exogenous CTGF and TGF-beta increased the rate of failure of GFS blebs. CONCLUSIONS: These data support the hypothesis that CTGF plays an important role in scarring and wound contracture after GFS. Inhibition of CTGF synthesis or its action may help prevent bleb failure and improve long-term GFS outcomes. PMID- 14744890 TI - Detecting early to mild glaucomatous damage: a comparison of the multifocal VEP and automated perimetry. AB - PURPOSE: To gain better understanding of the relationship between abnormalities detected by the multifocal VEP (mfVEP) compared with those detected by static achromatic, automated perimetry in patients with glaucoma. METHODS: Fifty patients were studied who had open-angle glaucoma that met the following criteria: (1) a mean deviation (MD) of better than -8 dB in both eyes on the 24-2 Humphrey visual field (HVF) test (Carl Zeiss Meditec, Dublin, CA); and (2) glaucomatous damage in at least one eye, as defined by a glaucomatous optic disc and an abnormal 24-2 HVF test result (pattern standard deviation [PSD] <5% and/or glaucoma hemifield test [GHT] results outside normal limits). Monocular mfVEPs were obtained from each eye by using a pattern-reversal dartboard array, 44.5 degrees in diameter, which contained 60 sectors. Recording electrodes were placed at the inion (I) and I+4 cm, and also at two lateral locations up 1 cm and over 4 cm from I. Monocular and interocular mfVEP probability plots were derived by comparing the results with those of normal control subjects. For both the HVF and mfVEP probability plots, a hemifield was classified as abnormal if three or more contiguous points were significant at less than 5%, with at least one at less than 1%. RESULTS: Of the 200 hemifields tested (50 patients x two eyes x two hemifields), 75 showed significant clusters on the HVF, and 74 (monocular probability plot) and 93 (monocular or interocular plot) showed significant clusters on the mfVEP. Overall, the HVF and mfVEP results agreed on 74% of the hemifields, and 90 hemifields were normal and 58 were abnormal on both the mfVEP (interocular and/or monocular abnormal) and HVF cluster tests. Of the 52 disagreements, 35 hemifields had a significant cluster on the mfVEP, but not on the HVF, whereas the reverse was true of 17 hemifields. A case-by-case analysis indicated that misses and false-positive results occurred on both the HVF and mfVEP tests. CONCLUSIONS: As predicted from a theoretical analysis, under these conditions (i.e., the signal-to-noise level) the HVF and monocular mfVEP tests showed a comparable number of defects, and, with the addition of the interocular test, the mfVEP showed more abnormalities than the HVF. However, although there were abnormalities detected by the mfVEP that were missed by the HVF, the reverse was true as well. PMID- 14744891 TI - Simultaneous comparison of relative damage to chromatic pathways in ocular hypertension and glaucoma: correlation with clinical measures. AB - PURPOSE: To use a new methodological approach, based on luminance noise, to assess without bias the relative damage of blue-yellow and red-green pathways in ocular hypertension and glaucoma and to correlate obtained measurements with clinical markers of disease progression. METHODS: A psychophysical procedure modified from Cambridge color test was used to assess color discrimination in three different groups: patients with primary open-angle glaucoma (n = 51 eyes), patients with ocular hypertension (n = 95 eyes), and control subjects (n = 46 eyes). Viewing conditions were such that the function of the macula was being tested, using a discrimination task under noisy conditions. Confusion vectors, and parameters obtained from discrimination ellipses were correlated with perimetric and clinical data taken from the same groups. RESULTS: The lengths of the major axis of chromatic discrimination ellipses and all confusion vectors were significantly different between the groups. These measures followed a significant gradient of worsening performance from the control to hypertensive and glaucoma groups, which was steeper for the tritan axis. There was a significant global positive correlation between test parameters and cup-to-disc ratio and a significant negative correlation with the perimetric mean deviation index. Ellipse length in patients with ocular hypertension correlated significantly with the duration of their hypertensive state. CONCLUSIONS: This psychophysical test can detect visual dysfunction in a significant subset of patients with long-term hypertension with preserved visual fields. Macular function is damaged earlier than previously believed, in both the blue-yellow and red-green pathways. PMID- 14744892 TI - Improved protection from primary ocular HSV-1 infection and establishment of latency using multigenic DNA vaccines. AB - PURPOSE: To compare the effectiveness of immunization with "naked" DNA corresponding to the genes encoding five HSV-1 glycoproteins, gB, gC, gD, gE, and gI (5gP DNA), with immunization with the five glycoproteins (5gP protein). Also, to compare immunization of 5gP protein in Montanide ISA 720 (SEPPIC, Paris, France), an adjuvant recently approved for use in humans, with immunization of 5gP protein in Freund's adjuvant. METHODS: BALB/c mice were vaccinated with 5gP DNA or 5gP protein emulsified in ISA 720 or Freund's adjuvant. Neutralizing antibody titers were determined by plaque-reduction assays. IL-2, -4, and -12 and IFN-gamma levels were determined by ELISA after in vitro stimulation of spleen cells. After ocular challenge with 2 x 10(5) plaque-forming units [pfu] per eye of HSV-1 strain McKrae, virus replication in the eye, survival, blepharitis, corneal scarring, and latency were determined. RESULTS: Neutralizing antibody titers (approximately 1:800-1:1200), corneal scarring (trace) and survival (100%) were similar for all vaccine groups, including 5gP DNA. Compared with the other vaccine groups, the 5gP DNA group had less ocular virus replication, as judged both by maximum virus titer and time of viral clearance. ISA 720 appeared more effective than Freund's against ocular virus replication and eye disease. The 5gP DNA-vaccinated mice had less blepharitis and latency than any other group and had the highest levels of IL-12 and IFN-gamma. All vaccine groups had similar levels of IL-2. CONCLUSIONS: The 5gP DNA vaccine appeared to be more effective than the corresponding protein subunit vaccine, regardless of adjuvant. Emulsification of the 5gP protein in ISA 720 appeared to be more effective than emulsification in Freund's adjuvant. PMID- 14744893 TI - Effects of TGF-beta2 on immune response-related gene expression profiles in the human corneal endothelium. AB - PURPOSE: To determine the effects of transforming growth factor (TGF)-beta2 on immune-response-related gene expression profiles in the stimulated human corneal endothelium (HCE). METHODS: A human complementary DNA (cDNA) expression array analysis was used to investigate the effects of TGF-beta2 on cultured 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 expression patterns revealed by the cDNA expression array analysis. Moreover, the expression of newly identified genes in HCE was confirmed by RT-PCR in human donor corneas. RESULTS: cDNA expression array analysis and semiquantitative RT-PCR revealed that TGF-beta2 downregulated the expression of IL-6, growth-related (Gro)-alpha (CXCL1), monocyte chemotactic protein (MCP)-1 (CCL2), granulocyte-colony stimulating factor (G-CSF), and insulin-like growth factor binding protein (IGFBP)-5 and upregulated the expression of tissue inhibitor of metalloproteinase (TIMP)-1. ELISA confirmed TGF-beta2-mediated changes in the expression of IL-6, CXCL1, CCL2, G-CSF, IGFBP-5, and TIMP-1 at the protein level. CXCL1, G-CSF, and IGFBP-5 mRNAs were detected for the first time in the HCE of donor corneas. CONCLUSIONS: TGF-beta2 downregulates IL-6, CXCL1, CCL2, G-CSF, and IGFBP-5, and upregulates TIMP-1 in cultured HCE stimulated with proinflammatory cytokines, suggesting the immunomodulatory role of TGF-beta2 in the aqueous humor and the pathophysiological significance of TGF-beta2 in the anterior chamber of the eye. PMID- 14744894 TI - Pseudomonas keratitis: protease IV gene conservation, distribution, and production relative to virulence and other Pseudomonas proteases. AB - PURPOSE: To determine the distribution of the protease IV gene, the production of this and other proteases by multiple strains of Pseudomonas, and the virulence of a mutant specifically deficient in protease IV. METHODS: The protease IV gene was cloned, its sequence analyzed, and its chromosomal location determined by pulse field gel electrophoresis. Three PCR reactions were used to detect the protease IV gene in 30 Pseudomonas isolates and protease production was determined by Western blot analysis, colorimetric assay, and zymography. An allelic replacement mutant deficient in the protease IV gene was analyzed for enzyme production, corneal growth, and corneal virulence. RESULTS: The protease IV gene was identified in all P. aeruginosa, but none of the non-aeruginosa strains tested. The protease IV genes of strains PA103-29 and PAO1 were in a common chromosomal site and had 98.5% sequence identity with variations occurring mainly in the promoter region. The protease IV activity of the 23 wild-type P. aeruginosa strains tested varied from 2.3 to 221.5 x 10(-3) U/mg protein in the culture supernatant. Protease IV was produced by all P. aeruginosa wild-type strains. A protease IV-deficient mutant derived from strain PA103-29 had reduced virulence compared with its parent strain and unexpectedly produced alkaline protease. CONCLUSIONS: The protease IV gene and its product are common to P. aeruginosa, but not to other Pseudomonas species. Protease IV activity varies among P. aeruginosa strains, and a mutant specifically deficient in this activity produced alkaline protease and had reduced corneal virulence. PMID- 14744895 TI - Macular pigment measurement by heterochromatic flicker photometry in older subjects: the carotenoids and age-related eye disease study. AB - PURPOSE: To develop a standardized protocol for measuring macular pigment optical density (MPOD) of experimentally naive subjects by heterochromatic flicker photometry (HFP). METHODS: MPOD in eyes of 54 women, age 50 and 79 years (mean, 66), was studied. The spatial profile of MPOD was measured in the right eye, and two spatial points were also measured in the left eye. Forty-eight of these inexperienced subjects completed the protocol on two separate visits. For a subset of the group, the MPOD at two different wavelengths was measured. RESULTS: The test-retest correlation at 0.5 degrees eccentricity in the right eye was 0.9. On the second visit, more than 90% of the subjects were able to perform the HFP test with results that were consistent with the absorption spectrum of macular pigment. On the first visit, data from the inexperienced subjects deviated more from the expected relationships between the two wavelengths, presumably because they had less skill in performing the task. However, subjects with high or low macular pigment density were distinguished clearly. CONCLUSIONS: Reliable and meaningful measurements of macular pigment density in older subjects can be made using HFP, with a standardized protocol in the limited time available in large epidemiologic studies. This protocol will be made freely available to other researchers on request. PMID- 14744896 TI - Magnetic resonance imaging study of the effects of age and accommodation on the human lens cross-sectional area. AB - PURPOSE: To evaluate the effect of age and accommodation on lens cross-sectional area (CSA). METHODS: High-resolution magnetic resonance images of the eye were acquired from 25 subjects ranging in age from 22 to 50 years during accommodation and with accommodation at rest. The images were analyzed to obtain the total lens CSA and the CSAs of the anterior and posterior portions of the lens. RESULTS: The total lens CSA and the CSA of the anterior portion increased with age in both accommodative states. With accommodation, the CSA was larger in these portions of the lens; however, this difference decreased with age. Conversely, the CSA of the posterior portion of the lens remained statistically independent of both age and accommodative state. CONCLUSIONS: This preliminary study documents, in vivo, that the lens grows with age. This growth appears to be confined to the anterior portion. A quite unexpected finding is that both the total lens CSA and the CSA of the anterior portion are greater during accommodation when zonular tension is minimized. This accommodative change in CSA, which decreases with age, may be due to compression of the lens material during relaxed accommodation when zonular tension is greatest. That both age and accommodative changes in CSA appear to be limited to the anterior portion of the lens may be related to properties of the anterior capsule and lens material, the position of the zonular attachments, and the location of the fetal nucleus. PMID- 14744897 TI - Posture changes and subfoveal choroidal blood flow. AB - PURPOSE: To evaluate the effect of posture change on subfoveal choroidal blood flow (ChBF) in normal volunteers. METHODS: The pulsatile, nonpulsatile, and mean ChBF were measured with laser Doppler flowmetry in 11 healthy volunteers with a mean age of 32 +/- 13 (SD) years. The posture of the subjects was changed from standing (90 degrees ), to supine (-8 degrees ), and back to standing, with a mechanically driven table. During the whole experimental procedure, ChBF and heart rate (HR) were continuously recorded. After 30 seconds in standing position, the subjects were tilted to supine during approximately 30 seconds. They remained in this position for approximately 2 minutes, after which they were tilted back to the standing position (recovery), where they remained for another approximately 2 minutes. Systemic brachial artery blood pressure (BP) was measured in the baseline, supine, and recovery positions. This procedure was repeated to measure the intraocular pressure (IOP) at the different postures. RESULTS: Mean BP did not change significantly throughout the experimental procedure. As the body was tilted from standing to supine, HR decreased by 16% (P < 0.0004), IOP increased by 29% (P < 0.001), and mean ChBF increased by 11% (P < 0.01). The increase in ChBF was primarily due to an increase in the nonpulsatile component of the blood velocity. CONCLUSIONS: Based on previously reported experimental data that indicate that the ocular perfusion pressure increases less than predicted by purely hydrostatic considerations when the body is tilted from the standing to the supine position, the observed increase in ChBF suggests a passive response of the choroidal circulation to the posture change. PMID- 14744898 TI - Influence of adrenomedullin on tone of isolated bovine retinal arteries. AB - PURPOSE: To assess and characterize the vasorelaxing effect of adrenomedullin (AM) on isolated bovine retinal arteries (BRAs). METHODS: Retinal arteries were isolated from bovine eyes and mounted in a wire myograph for isometric tension recording. Concentration-response curves were generated by cumulative addition of AM (1 pM to 0.1 micro M) to the organ bath. RESULTS: AM caused a concentration dependent relaxation of the BRAs. Removal of the endothelium of the BRAs, inhibition of nitric oxide synthase with -nitro-L-arginine (L-NA) or inhibition of soluble guanylyl cyclase with 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ) significantly reduced the AM response. Cyclooxygenase inhibition with indomethacin or sodium diclofenac did not reduce, but rather increased, vasodilation. The AM-receptor antagonist AM 22-52 slightly, but significantly, reduced the AM response, whereas the CGRP-receptor antagonist CGRP 8-37 caused a more pronounced reduction. The adenosine receptor antagonist 8-(p-sulfophenyl) theophylline (8-SPT) did not affect AM-induced vasorelaxation. Inhibition of several intracellular calcium ([Ca(2+)](i))-reducing mechanisms failed to block the relaxation induced by AM. Only inhibition of the plasma membrane Ca(2+) adenosine triphosphatase (ATPase) with vanadate significantly attenuated the AM response. CONCLUSIONS: AM induces vasodilation in isolated bovine retinal arteries. Endothelium-derived NO and stimulation of CGRP- and AM-receptors appear to be involved in the AM response, whereas prostanoids and activation of adenosine receptors are not involved. Activation of Ca(2+)-extrusion by the plasma membrane Ca(2+)-ATPase may elicit the relaxation of BRAs in response to AM. PMID- 14744899 TI - Electrophysiological studies of the feasibility of suprachoroidal-transretinal stimulation for artificial vision in normal and RCS rats. AB - PURPOSE: Assessment of a novel method of retinal stimulation, known as suprachoroidal-transretinal stimulation (STS), which was designed to minimize insult to the retina by implantation of stimulating electrodes for artificial vision. METHODS: In 17 normal hooded rats and 12 Royal College of Surgeons (RCS) rats, a small area of the retina was focally stimulated with electric currents through an anode placed on the fenestrated sclera and a cathode inserted into the vitreous chamber. Evoked potentials (EPs) in response to STS were recorded from the surface of the superior colliculus (SC) with a silver-ball electrode, and their physiological properties and localization were studied. RESULTS: In both normal and RCS rats, STS elicited triphasic EPs that were vastly diminished by changing polarity of stimulating electrodes and abolished by transecting the optic nerve. The threshold intensity (C) of the EP response to STS was approximately 7.2 +/- 2.8 nC in normal and 12.9 +/- 7.7 nC in RCS rats. The responses to minimal STS were localized in an area on the SC surface measuring 0.12 +/- 0.07 mm(2) in normal rats and 0.24 +/- 0.12 mm(2) in RCS rats. The responsive area corresponded retinotopically to the retinal region immediately beneath the anodic stimulating electrode. CONCLUSIONS: STS is less invasive in the retina than stimulation through epiretinal or subretinal implants. STS can generate focal excitation in retinal ganglion cells in normal animals and in those with degenerated photoreceptors, which suggests that this method of retinal stimulation is suitable for artificial vision. PMID- 14744900 TI - Changes in focal macular ERGs after macular translocation surgery with 360 degrees retinotomy. AB - PURPOSE: To evaluate the short- and long-term changes of focal macular electroretinograms (fmERGs) after macular translocation with 360 degrees retinotomy. METHODS: This was a retrospective study. fmERGs were recorded in 19 eyes of 19 consecutive patients who underwent macular translocation with 360 degrees retinotomy for choroidal neovascularization (CNV) secondary to age related macular degeneration (AMD; 17 eyes) or polypoidal choroidal vasculopathy (2 eyes). The changes in the fmERGs, recorded before, shortly after (6-12 months; mean 8.3 months), and more than 18 months (18-30 months; mean 22.4 months) after surgery from 12 eyes, were analyzed. A 15 degrees stimulus centered on the fovea was used to elicit the fmERGs. RESULTS: The mean logarithm of minimum angle of resolution (logMAR) was 1.06 +/- 0.07 (20/230) before surgery, 0.78 +/- 0.08 (20/121) early after surgery (n = 19), and 0.64 +/- 0.07 (20/87) late after surgery (n = 12). These improvements in visual acuity were significant (P = 0.0074, P = 0.0050, respectively). Before surgery, the amplitudes of all components of the fmERGs were markedly reduced in all eyes. The mean b-wave amplitude in 17 AMD eyes recorded early after surgery was significantly larger (P = 0.0262), and the mean a-wave amplitude was also increased but not significantly (P = 0.1180). The mean amplitudes of the a- and b-waves in 10 AMD eyes recorded after 18 months were significantly larger than those before the surgery (P = 0.0218, and P = 0.0284). The mean implicit time of the b-wave in 17 AMD eyes decreased early after surgery, and a further decrease was detected at the later testing time. CONCLUSIONS: These results indicate that macular function is partially recoverable after macular translocation in some patients. PMID- 14744901 TI - Photopic and scotopic fine matrix mapping of retinal areas of increased fundus autofluorescence in patients with age-related maculopathy. AB - PURPOSE: To investigate photopic and scotopic sensitivity of retinal areas that show increased fundus autofluorescence (FAF) in patients with age-related maculopathy (ARM). METHODS: FAF was imaged with a modified confocal scanning laser ophthalmoscope (cSLO). Fine matrix mapping (FMM) was performed with a modified field analyzer. Photopic and scotopic thresholds were obtained at 100 locations on a 9 degrees x 9 degrees matrix with 1 degrees spacing, centered on a macular area of increased FAF. Inclusion criteria included ARM fundus changes, areas of increased FAF, central and stable fixation, and visual acuity of 20/40 or better. RESULTS: FAF images were reviewed in 436 patients with age-related maculopathy (ARM), of whom 38 met the inclusion criteria. FMM was performed in seven eyes of seven patients. Areas of increased FAF in patients with late ARM (choroidal neovascularization or geographic atrophy) showed normal or only mildly abnormal photopic, but severely reduced scotopic, sensitivity. The central area of increased FAF corresponding to a large foveal druse in a patient with ARM showed moderately reduced photopic and severely reduced scotopic sensitivity. In the other patients with ARM with drusen, areas of increased FAF showed normal or near-normal photopic sensitivity, but moderately reduced scotopic sensitivity. CONCLUSIONS: In retinal areas of increased FAF in patients with ARM, scotopic sensitivity loss considerably exceeded photopic sensitivity loss. This finding is in line with histologic data that have demonstrated a preferential loss of rods in ARM, but does not explain the magnitude of sensitivity loss. The study shows that increased FAF in ARM has a functional correlate. PMID- 14744902 TI - Retinal function loss after monocarboxylate transport inhibition. AB - PURPOSE: To test the proposal that inhibiting monocarboxylate transport in the rat retina results in altered retinal function measured using the electroretinogram (ERG) and to evaluate the efficacy of exogenous metabolic substrates to restore any functional deficit. METHODS: Full-field white-flash ERGs were measured after monocarboxylate transport inhibition with intravitreal injection of alpha-cyano-4-hydroxycinnamic acid (4-CIN, 10 mM), and functional recovery was assessed after the introduction of various exogenous metabolic substrates (10 mM): lactate, pyruvate, alpha-ketoglutarate, alanine, succinate, and glutamine. The efficacy of glutamine as a metabolic substrate was also considered in the presence of phosphate-activated glutaminase inhibition (6-diazo 5-oxo-norleucin, 10 mM) or aminotransferase inhibition (aminooxyacetic acid, 10 mM). Pyruvate and alanine recovery was also assessed after aminooxyacetic acid application. RESULTS: 4-CIN application resulted in an increased phototransduction amplitude but a mild reduction of gain. A greater reduction of postreceptoral b-wave and oscillatory potential amplitudes (80%) was observed, along with delayed implicit times (35 ms). Partial recovery of b-wave amplitudes was achieved with exogenous lactate (24%), pyruvate (27%), alpha-ketoglutarate (27%), alanine (25%), and succinate (26%), whereas glutamine provided 62% recovery. However, none of the substrates improved phototransduction gain. Both 6 diazo-5-oxo-norleucin and aminooxyacetic acid completely suppressed the glutamine induced b-wave recovery. Aminooxyacetic acid also abolished the b-wave recovery from 4-CIN afforded by pyruvate and alanine. CONCLUSIONS: The greater loss of the b-wave and oscillatory potentials may reflect preferential routing of amino acid carbon skeletons to oxidative metabolic pathways, which in turn reduces glutamate availability for neurotransmission between photoreceptors and ON-bipolar cells. The reduction in log S provides evidence that inhibition of monocarboxylate transport produced some metabolic dysfunction in the rat. PMID- 14744903 TI - Microvascular remodeling after occlusion-recanalization of a branch retinal vein in rats. AB - PURPOSE: To describe the time course of microvascular changes after transient branch retinal vein occlusion (BRVO) in rats. METHODS: BRVO was induced in pigmented rats by focal laser photocoagulation. The subsequent changes in the retinal angiogram were followed up, both in vivo by confocal scanning laser ophthalmoscopy and ex vivo by confocal microscopy. RESULTS: At day 1, capillary closure affected the three microvessel layer differentially, the intermediary layer being the most affected. Collateral veins, which were initiated by the dilation of deep-layer venules, pursued their course below adjacent arteries. These microvascular changes peaked between days 1 and 3. After recanalization at day 3, microvascular changes regressed gradually but incompletely, and at day 30 capillary closure and venule dilation persisted. CONCLUSIONS: Transient occlusion of a retinal vein in rats leads to short- and long-term microvascular remodeling upstream of the occlusion site. This study describes a model for the tridimensional arrangement of retinal microvessel that accounts for the topography of the early capillary closure and collateral vessel formation that occur after BRVO. In the long term, these changes regressed incompletely, with recanalization of the occluded vein, suggesting that after a short period of occlusion, microvascular changes may become at least partially independent of flow. Despite the intrinsically limited applicability of this model to human vein occlusion, the results suggest that even if therapeutic decompression of an occluded vein is performed early, it may not reverse capillary dropout completely. PMID- 14744904 TI - Electroretinography as a screening method for mutations causing retinal dysfunction in mice. AB - PURPOSE: To detect mice with hereditary retinal impairment, a high-throughput electroretinography (ERG) screening system was established. METHOD: Mice from eight different strains without known retinal disorders (102, 129/SvJ, AKR, C57BL/6J, C57BL/6JIco, CBA/CaJ, and DBA/2NCrlBR) and one control strain with retinal degeneration (C3HeB/FeJ) were fixed on a specially constructed sled, ERG electrodes were placed on the cornea, and mice were moved into a Ganzfeld stimulator. From a luminance range of 0.0125 to 500 cd-s/m(2) in a pretest series two levels (5 and 125 cd-s/m(2)) were chosen to shorten examination times. The root mean square (RMS) of the ERG-recording was analyzed to detect animals with abnormal retinal function. ERG responses of the left and right eyes were compared in amplitudes and implicit times of the a- and b-waves. Statistical analysis of the latter parameters was performed in all wild-type animals. Histology was performed on selected mice. RESULTS: ERG recordings of individual animals for the left and right eye revealed good agreement in amplitudes and implicit times of the a- and b-waves (P < 0.05). Comparison of these parameters among the wild-type strains showed several differences. Evaluation of the RMS revealed, in addition to the C3HeB/FeJ mice, a subgroup of mice within the 129/SvJ strain with abnormal retinal function. Molecular analysis of these mice demonstrated the presence of the same retroviral insertion in the Pde6b gene, which is causative of the Pde6b(rd1) allele carried in C3HeB/FeJ mice. Histologic analysis demonstrated good correlation between retinal electrophysiology and morphology. CONCLUSIONS: The present results demonstrate the feasibility of ERG for screening a large number of mice to detect animals with functional retinal impairment. PMID- 14744905 TI - Comparison of Heidelberg Retina Tomograph II and Retinal Thickness Analyzer in the assessment of diabetic macular edema. AB - PURPOSE: To compare the within-session variability of the Macular Edema Module of the Heidelberg Retinal Tomograph II (HRT II; Heidelberg Engineering, Heidelberg, Germany) and the Retinal Thickness Analyzer (RTA, Talia Technology Ltd., Neve Ilan, Israel) in patients with diabetes and nondiabetic subjects and to determine the agreement of both instruments to clinical observation. METHODS: Seventeen patients with diabetic macular edema (DME) and 17 nondiabetic subjects were examined. Three scans of the posterior pole were acquired for each volunteer with both the HRT II and the RTA. The edema index and foveal average thickness were derived for a 600- micro m radius circle centered on the fovea for the HRT II and RTA scans. The coefficient of variation (COV) was calculated. Clinical examination of detectable edema was performed using stereo fundus biomicroscopy and the level of agreement between each instrument and clinical observation was determined using a zonal analysis. RESULTS: Individual COVs for the HRT II and RTA ranged from 2.3% to 24.6% (median 8.3%) and 2.1% to 46.7% (median 6.4%), respectively, in diabetic subjects and 2.0% to 37.5% (median 6.0%) and 2.3% to 14.7% (median 8.5%), respectively, in nondiabetic subjects. Clinical assessment identified edema in a total of 47 sectors in patients with DME. In comparison to clinical assessment, the HRT II gave a sensitivity of 92% and a specificity of 68% and the RTA gave a sensitivity of 57% and a specificity of 71%. CONCLUSIONS: Both instruments have good within-session repeatability. The HRT II showed better agreement with clinical assessment than the RTA. The agreement between instruments was poor. PMID- 14744906 TI - Supplemental oxygen improves diabetic macular edema: a pilot study. AB - PURPOSE: Diabetic macular edema (DME) is the most common cause of moderate visual disability in persons of working age in the United States. The pathogenesis of DME is poorly understood. In this study, the effect of retinal hypoxia in the development and maintenance of DME was investigated. METHODS: Five patients with chronic DME despite at least one focal laser photocoagulation treatment (nine eyes) received 4 L/min of inspired oxygen by nasal cannula for 3 months. Best corrected visual acuity (VA) and retinal thickness, assessed by optical coherence tomography (OCT), were measured at baseline, during 3 months of oxygen treatment, and for 3 months after stopping oxygen. RESULTS: After 3 months of oxygen therapy, nine of nine eyes with DME at baseline showed a reduction in thickness of the center of the macula. Foveal thickness (FTH) above the normal range was reduced by an average of 43.5% (range, 14%-100%), excess foveolar thickness (CEN) was reduced by an average of 42.1% (range, 13%-100%), and excess macular volume was reduced by an average of 54% (range, 35%-100%). Statistical analyses suggested that these changes were unlikely to be due to chance (P = 0.0077 by Wilcoxon signed-rank test). Three eyes showed improvement in VA by at least 2 lines, one by slightly less than 2 lines, and five eyes showed no change. Three months after discontinuation of oxygen, five of the nine eyes showed increased thickening of the macula compared with when oxygen was discontinued. CONCLUSIONS: Supplemental inspired oxygen may decrease macular thickness due to DME, suggesting that retinal hypoxia is involved in the development and maintenance of DME. PMID- 14744907 TI - The squirrel monkey: characterization of a new-world primate model of experimental choroidal neovascularization and comparison with the macaque. AB - PURPOSE: To evaluate and characterize the New-World squirrel monkey as a primate model for experimental choroidal neovascularization (CNV) studies and to compare it with the current Old-World macaque monkey model. METHODS: Fibrovascular tissues (FVT) were elicited in 12 maculae of seven squirrel monkeys by laser photocoagulation using optimized laser parameters (532 nm, 0.05 second, 75 micro m, 650 mW). Follow-up fundus and fluorescein angiography (FA) examinations were conducted on postlaser days 30 and 35, followed by euthanasia and histologic analysis of tissues. For comparative evaluations, FVT development also was induced and analyzed in eight maculae of four macaque monkeys with laser parameters previously used in this species (514 nm, 0.1 second, 50 micro m, 390 and 455 mW). RESULTS: FVT developed in both primate species, consisting of fibrous tissue that contained vessels that ranged from sparse but identifiable capillaries to well-established neovascular networks. Overall, 65% of the photocoagulation sites in the squirrel monkey and 37% of sites in macaque monkey elicited development of FVT. Localized FVT ranged from modest to extensive thickenings of the choriocapillaris layer. Unexpectedly, 76% of the FVT sites in squirrel monkey eyes and 27% of the sites in macaque eyes showed diffuse FVT that expanded beyond the original photocoagulation sites, accompanied by neovascular infiltration of the retina. CONCLUSIONS: Like the macaque, the squirrel monkey can be considered a useful primate model for experimental CNV investigations, while additionally offering certain species-specific advantages. Diffuse FVT permit studies of antiangiogenic therapies in areas distant from laser photocoagulative trauma sites. PMID- 14744908 TI - Inhibition of choroidal neovascularization in rats by the urokinase-derived peptide A6. AB - PURPOSE: To evaluate the inhibitory effects of a urokinase-derived octapeptide A6 on laser-induced choroidal neovascularization (CNV). METHODS: In the first arm of the study, subcutaneous injection of A6 (200 mg/kg per day) into the right eyes in 20 rats and phosphate-buffered saline in 20 control rats was started 1 day before laser injury. Angiography was performed at week 2. To evaluate the dose response, a second arm of the study was performed in the left eyes. Half of the treatment group was treated with 400 mg/kg per day, and the remaining half continued to receive 200 mg/kg per day beginning on week 4. Laser injury was made at week 5 and angiography was performed at week 7. Angiographic evaluation, histopathologic evaluation including maximum CNV thickness and factor-VIII stained endothelium counting were performed in the second arm of the study. Choroidal concentrations of A6 were measured. RESULTS: In the first arm of the study, angiography showed a 40.8% reduction in CNV in the 200-mg/kg per day treatment group, compared with the control (P = 0.0008). In the second arm of the study, angiographic reduction in CNV was 37.9% in the 200-mg/kg per day group (P = 0.0314) and 70.0% in the 400-mg/kg per day group (P = 0.0124), compared with the control. CNV was significantly less in the 400-mg/kg per day group than in the 200-mg/kg per day group (P = 0.0393). Both CNV thickness and number of endothelial cells were reduced in a dose-dependent manner and significantly less than in the control (P < 0.05). Mean choroidal concentration of A6 2 hours after injection was 0.72 micro M in the 200-mg/kg per day (100 mg/kg every 12 hours) and 1.75 micro M in the 400-mg/kg per day (200 mg/kg every 12 hours) treatment groups. Levels at 11 hours after injections were undetectable. CONCLUSIONS: A6 demonstrated antiangiogenic properties in a rat model of CNV and may be useful in the treatment of CNV. PMID- 14744909 TI - Posterior vitreous detachment induced by microplasmin. AB - PURPOSE: To demonstrate the efficacy of microplasmin in inducing posterior vitreous detachment (PVD) and to evaluate the human and the feline retina after treatment. METHODS: Thirteen human donor eyes were injected with 62.5, 125, or 188 micro g microplasmin. The 13 fellow eyes received balanced salt solution. Four of the microplasmin-treated eyes received an additional intravitreal gas injection. After incubation at 37 degrees C for 30 minutes, all globes were placed in 4% paraformaldehyde. Retinal specimens were processed for scanning (SEM) and transmission (TEM) electron microscopy. Five feline eyes were injected with 14.5- or 25- micro g microplasmin. Animals were killed after 1 day, 3 days, or 3 weeks, and retinal specimens were evaluated by electron and confocal microscopy. RESULTS: In all control eyes, SEM demonstrated the cortical vitreous covering the inner limiting membrane (ILM). Intravitreal injection of 125 or 188 micro g microplasmin resulted in complete PVD. After treatment with 62.5 micro g microplasmin, SEM revealed collagen fibrils covering the ILM. Additional gas injection did not change the dose necessary for PVD. In vivo in cats, 25 micro g microplasmin resulted in complete PVD after 3 days. After 3 weeks, there was complete PVD with both doses of microplasmin. The retina and the ILM were well preserved in all eyes. CONCLUSIONS: Both after death and in vivo, microplasmin induces a dose-dependent cleavage between the vitreous cortex and the ILM without morphologic alterations of the retina. In the feline eye, there is no cellular response of retinal glial cells or neurons. PMID- 14744910 TI - Preventing retinal detachment-associated photoreceptor cell loss in Bax-deficient mice. AB - PURPOSE: To characterize photoreceptor cell apoptosis and cell loss in a mouse model of experimental retinal detachment (RD), and to use the technology of mouse genetics to study the molecular mechanisms underlying RD-associated photoreceptor degeneration. METHODS: Retinal detachments were created in adult wild-type and Bax-deficient mice by subretinal injection of 1.4% sodium hyaluronate. At 1, 3, 7, and 28 days after injection, animals were killed, eyes enucleated, and retinal sections studied by histochemistry, immunofluorescence labeling, and confocal microscopy. Rods and cones were labeled, and apoptotic cells were identified with terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL). Photoreceptor cell apoptosis and cell loss were assessed quantitatively by counting both surviving and TUNEL-positive rods and cones. RESULTS: TUNEL positive cells were found within the outer nuclear layer (ONL) of the detached portions of the retina. They were detected in the detached retina on day 1, peaked on day 3, and dropped precipitously after day 7 after RD. Photoreceptor cell loss of both rods and cones followed a similar time course after RD. Moreover, deletion of the proapoptotic gene Bax in a knockout mouse model abolished the RD-associated photoreceptor cell degeneration. CONCLUSIONS: Apoptosis is a major mechanism leading to photoreceptor cell death after RD. Blockage of the activity of the proapoptotic molecule Bax in a knockout mouse model prevents photoreceptor cell apoptosis and cell loss. These data suggest that the Bax-mediated apoptotic signaling pathway plays a critical role in RD associated photoreceptor cell death. PMID- 14744911 TI - Expression of CNTF receptor-alpha in chick violet-sensitive cones with unique morphologic properties. AB - PURPOSE: Application of ciliary neurotrophic factor (CNTF) can rescue mature photoreceptors from lesion-induced and hereditary degeneration. In the chick retina, expression of the CNTF receptor is present in a subpopulation of photoreceptor cells. The present study was undertaken to identify the CNTF receptor-expressing photoreceptors and to describe the subcellular localization of the receptor protein. METHODS: The localization of the CNTF receptor was analyzed by light and electron microscopic immunocytochemistry in chick retinal wholemount preparations, with an antibody for CNTF receptor alpha (CNTFRalpha). Immunoreactive cells were identified by double labeling with immunocytochemical markers for photoreceptor subpopulations. RESULTS: The CNTFRalpha antibody labeled evenly distributed outer segments (OS) of a photoreceptor subpopulation. CNTFRalpha-positive OS were associated with oil droplets of uniform size. Receptor immunoreactivity did not colocalize with markers for rods and red-green cones. Complete overlap was found after double labeling with the antibody CERN 933, which recognizes violet-sensitive cones in the chick retina. Ultrastructurally, the CNTFRalpha-immunoreactive OS showed rodlike properties: an elongated shape and stacks of membrane discs separated from the plasma membrane. Immunoreactivity was completely restricted to the plasma membrane of the OS and the inner membrane sheet of the photoreceptor calices present in avian retinas. CONCLUSIONS: CNTFRalpha expression identifies a unique type of photoreceptors in the avian retina which does not fit into the classic morphologic definition of rods and cones. The specific expression in violet-sensitive photoreceptors suggests that CNTF may have a neuroprotective role related to the specific function of these cells. PMID- 14744912 TI - Neuroprotective effect of inosine on axotomized retinal ganglion cells in adult rats. AB - PURPOSE: To explore the potential survival-promoting effect of inosine on axotomized retinal ganglion cells (RGCs) of adult rats in vivo. METHODS: The left optic nerves (ON) in the subject rats were transected at 1.5 mm from the optic disc. Repeated intraperitoneal injections or single intraocular injection of inosine were administered. The RGCs were retrogradely labeled with a gold fluorescent dye and the density of surviving RGCs in number per square millimeter of retina was calculated in wholemounted retinas. The functional integrity of the blood-retinal barrier (BRB) after ON transection was evaluated with an intravenous injection of Evans blue. RESULTS: In control animals, the mean density of surviving RGCs (number per square millimeter) of the whole retina was 2007 +/- 68 at 2 days (taken as the normal value), 927 +/- 156 at 7 days, and 384 +/- 33 at 14 days after surgery. Repeated intraperitoneal injections (75 mg/kg for each injection) of inosine significantly enhanced RGC survival at 14 days after ON transection (500 +/- 38), whereas no significant difference in the densities was detected at 7 days (974 +/- 101), even when the dosage of inosine was doubled (1039 +/- 61). At this time point, however, a single intraocular injection of inosine significantly increased the density of surviving RGCs (1184 +/- 156). Moreover, more RGCs around the optic disc were rescued when inosine, administered either intraperitoneally or intraocularly, showed a beneficial effect on RGC survival. No breakdown of the BRB after ON transection was detected with the method used in the study. CONCLUSIONS: These findings demonstrate that inosine could protect axotomized RGCs in vivo after ON transection. PMID- 14744913 TI - Involvement of rho-kinase pathway in contractile activity of rabbit RPE cells in vivo and in vitro. AB - PURPOSE: Increased alpha-smooth muscle actin (alpha-SMA) expression in epiretinal membranes causes tractional retinal detachment (TRD) in proliferative vitreoretinopathy (PVR). The Rho-A/Rho-associated kinase signaling pathway is a principal mediator of contractile force generation in nonmuscle cells. In the current study, the relation between the Rho-kinase pathway and alpha-SMA expression and type I collagen gel contractile activity in retinal pigment epithelial (RPE) cells was investigated, using Y27632, a specific inhibitor of p160ROCK, and the involvement of the Rho-kinase pathway was evaluated in a rabbit PVR model with cultured RPE cells and platelet-rich plasma (PRP). METHODS: RPE cells were obtained from rabbits and cultured. The number of passages and the effect of Y27632 on alpha-SMA expression were studied by immunohistochemistry and immunoblot analysis. An in vitro type I collagen gel contraction assay and MTT assay evaluated the effect of Y27632 on RPE cell contractile force and proliferation. Cultured sixth-passage rabbit RPE cells were coinjected with PRP intravitreally, followed by 50 micro M of Y27632, injected weekly. The presence of TRD was assessed until 28 days to evaluate the effect of Y27632 in vivo. RESULTS: Expression of alpha-SMA was increased according to the passages. Y27632 suppressed alpha-SMA expression in cultured RPE cells and impaired contractile force. Y27632 did not affect the proliferative potential. Y27632 significantly (P < 0.01) reduced TRD development. CONCLUSIONS: Y27632 decreased alpha-SMA expression and the contractile force generated by RPE cells and attenuated PVR, indicating the involvement of the Rho-kinase pathway in cell-dependent collagen contraction in vitro and in vivo. The drug may affect the biological event by inhibiting alpha-SMA expression, and Y27632 could be useful for preventing PVR. PMID- 14744915 TI - The effect of retinal ganglion cell injury on light-induced photoreceptor degeneration. AB - PURPOSE: To determine the effect of optic nerve transection (ONT) and excitotoxic retinal ganglion cell (RGC) injury on light-induced photoreceptor degeneration. METHODS: Age- and sex-matched rats underwent unilateral ONT or received intravitreal injections of N-methyl-D-aspartate (NMDA). The fellow eye received sham treatment, and 7 or 21 days later each eye was subjected to an intense photic injury. Maximum a- and b-wave amplitudes of the flash electroretinogram (ERG) were measured at baseline, after the RGC insult, and 5 days after the photic injury. Semiquantitative reverse transcription-polymerase chain reaction analysis and immunoblot analysis were used to assess rod opsin mRNA and rhodopsin kinase protein levels and to measure defined trophic factors 7 or 21 days after ONT or injection of NMDA. Structural changes after the insults were determined histologically and immunohistochemically. RESULTS: ONT caused time-dependent reductions in the mean a- and b-wave amplitudes. Seven days after intravitreal NMDA the b-wave amplitude was reduced, but the a-wave was unaffected. ONT and NMDA injection attenuated the light-induced reductions in the a- and b-wave. Rod opsin mRNA levels and rhodopsin kinase protein levels were also significantly greater in the axotomized and NMDA-treated eyes compared with the sham-treated fellow eyes after the photic injury. Structural protection in the RGC-injured eyes was also evident histologically. Fibroblast growth factor (FGF)-2, ciliary neurotrophic factor (CNTF), and glial fibrillary acidic protein (GFAP) were significantly upregulated after ONT and NMDA. CONCLUSIONS: ONT and intravitreal injection of NMDA protect against subsequent photic injury. This protection may relate to the activation of retinal glial cells and the possible action of trophic factors such as FGF-2 and CNTF. PMID- 14744914 TI - Oxidative stress affects the junctional integrity of retinal pigment epithelial cells. AB - PURPOSE: Oxidative stress has been implicated in the pathogenesis of age-related macular degeneration. The cell line ARPE-19 was therefore examined for response to oxidative stress and its effect on stress protein induction and junctional integrity. METHODS: ARPE-19 cell viability after 1 week or 5 weeks in culture was assessed in response to different concentrations of hydrogen peroxide. The response to sublethal doses was assessed by examination of heme oxygenase (HO)-1, Hsp27 and Hsp70 by immunofluorescence and Western blot analysis. Immunofluorescence was used to investigate the localization of the junctional proteins zonula occludens (ZO)-1, occludin, and N-cadherin, and beta-catenin. Subcellular fractionation was used to assess any redistribution of beta-catenin. Monolayer integrity was examined by measurement of flux of rhodamine-conjugated dextrans from the apical to basal aspect of cells. RESULTS: ARPE-19 cells cultured for 5 weeks were less sensitive to chronic oxidative stress induced by hydrogen peroxide than those cultured for 1 week. The more differentiated ARPE-19 cells had higher steady state levels of Hsp27 and Hsp70. The response to stress also differed with time in culture. The localization of junctional proteins, which became strongly peripheral after 5 weeks in culture, became disrupted after oxidative stress, and cytosolic beta-catenin increased. Chronic oxidative stress also increased paracellular flux across the monolayer. CONCLUSIONS: Increased resistance to chronic oxidative stress with differentiation in ARPE-19 cells correlated with higher steady state levels of Hsp27 and Hsp70. Oxidative stress disrupted RPE cell junction and barrier integrity, which may contribute to the pathogenesis of diseases related to RPE through disruption of the blood-retinal barrier. PMID- 14744916 TI - Osmoregulation of taurine transporter function and expression in retinal pigment epithelial, ganglion, and muller cells. AB - PURPOSE: To determine whether taurine transporter (TauT) activity and expression are regulated by hyperosmolarity in RPE, ganglion, and Muller cells. METHODS: Uptake of taurine was measured in ARPE-19 cells cultured in DMEM-F12 medium without or with the addition of 50 mM NaCl or 100 mM mannitol. The kinetics of the transport were analyzed. RT-PCR and Northern and Western blot analyses were used to assess TauT mRNA and protein levels. The influence of hyperosmolarity on the uptake of taurine, myo-inositol, and gamma-aminobutyric acid GABA was studied in RPE, RGC-5, and rMC1 cells. RESULTS: TauT activity was abundant in RPE and was stimulated (3.5-fold) when the cells were exposed to hyperosmolar conditions (DMEM-F12 culture medium plus 50 mM NaCl or 100 mM mannitol). Peak stimulation of taurine uptake occurred after 17 hours of exposure to hyperosmolar medium. Kinetic analysis revealed that the hyperosmolarity-induced stimulation was associated with an increase in V(max) of TauT with no change in K(m). TauT mRNA and protein levels increased in RPE cells exposed to hyperosmolar conditions. Hyperosmolarity also stimulated the uptake of myo-inositol ( approximately 15 fold); GABA uptake was influenced less markedly. Immunofluorescence and functional studies showed that TauT is present in cultured RGC-5 and rMC1 cells. TauT activity was robust in these cells in normal osmolar conditions and increased by approximately twofold in hyperosmolar conditions. CONCLUSIONS: These studies provide the first evidence that hyperosmolarity regulates TauT activity and expression in RPE and that TauT is present in ganglion and Muller cells and is regulated by hypertonicity. The data are relevant to diseases such as diabetes, macular degeneration, and neurodegeneration, in which retinal cell volumes may fluctuate dramatically. PMID- 14744917 TI - Excitability changes induced in the human primary visual cortex by transcranial direct current stimulation: direct electrophysiological evidence. AB - PURPOSE: Transcranial direct current stimulation (tDCS) has been shown to modify the perception threshold of phosphenes elicited by transcranial magnetic stimulation (TMS). The current study was undertaken to examine whether tDCS, when applied over the occipital cortex, is also able to affect visual-evoked potentials (VEPs), which characterize occipital activation in response to visual stimulation, in a polarity-specific way. METHOD: For this purpose, VEPs evoked by sinusoidal luminance grating in an on/off mode were recorded before, immediately after, and 10, 20, and 30 minutes after the end of 5, 10, or 15 minutes of anodal or cathodal tDCS of the primary visual cortex. RESULTS: Significant effects were observed only when low-contrast visual stimuli were applied. Cathodal stimulation decreased, whereas anodal stimulation increased the amplitude of the N70 component. The effect of cathodal stimulation was significant immediately after and 10 minutes after the end of stimulation, if the stimulation duration was sufficiently long (i.e., 10-15 minutes). An increase of N70 amplitude by anodal stimulation was significant only 10 minutes after the end of the 15 minutes tDCS. Cathodal stimulation tended also to affect the amplitude of the P100 component; however, the effect of stimulation was inverse. The amplitude increased immediately after the end of cathodal stimulation. In contrast, anodal stimulation did not affect the P100. The latencies of the N70 and the P100 were not affected by tDCS. CONCLUSIONS: tDCS appears to be a suitable method of inducing reversible excitability changes in a polarity-specific way, not only in the motor but also in the primary visual cortex. The duration of the induced aftereffects depends not only on stimulation duration but also on stimulation polarity. Cathodal stimulation seems to be more effective, in line with previous reports on the motor cortex. PMID- 14744918 TI - Functional characterization of the L-type Ca2+ channel Cav1.4alpha1 from mouse retina. AB - PURPOSE: To study the electrophysiological and pharmacological properties of the L-type Ca(2+) channel (LTCC) Ca(v)1.4alpha1 (alpha1F) subunit from mouse retina and assess their contributions to the native retinal channel. METHODS: The full length cDNA of Ca(v)1.4alpha1 was cloned from murine retina in an RT-PCR approach. Ca(v)1.4alpha1 was expressed alone or together with the auxiliary alpha2delta1 and beta2a or beta3 subunits in HEK293 cells. The electrophysiological and pharmacological characteristics of L-type Ca(2+) and Ba(2+) inward currents (I(Ca) and I(Ba)) induced by Ca(v)1.4alpha1 were determined by the whole-cell configuration of the patch-clamp method and compared with currents induced by the cardiac and smooth muscle-type Ca(v)1.2alpha1 (alpha1C) channel. RESULTS: Ca(v)1.4alpha1-mediated I(Ba) was observed only when the alpha2delta1 and beta subunits were coexpressed. Current densities were approximately two times higher with beta2a than with beta3. I(Ba) activated faster and revealed much slower time-dependent inactivation than I(Ba) induced by Ca(v)1.2alpha1. Unlike in Ca(v)1.2alpha1, inactivation was not accelerated with Ca(2+) as the charge carrier, indicating the absence of Ca(2+)-dependent inactivation in Ca(v)1.4alpha1. Ca(v)1.4alpha1 exhibited voltage-dependent inactivation. The dihydropyridine (DHP) antagonist isradipine blocked Ca(v)1.4alpha1 with approximately 20-fold lower sensitivity than Ca(v)1.2alpha1. The agonistic DHP BayK 8644 stimulated maximum I(Ba) approximately sixfold. Ca(v)1.4alpha1 revealed only moderate sensitivities to L- and D-cis-diltiazem, with IC(50) in the micromolar range. Both enantiomers unexpectedly blocked Ca(v)1.4alpha1 with almost equal IC(50). CONCLUSIONS: The data indicate that Ca(v)1.4alpha1 subunit constitutes the major molecular correlate of retinal L type Ca(2+) current. Its intrinsic biophysical properties, in particular its unique inactivation properties, enable Ca(v)1.4alpha1 to provide a sustained I(Ca) over a voltage range such as required for tonic glutamate release at the photoreceptor synapse. PMID- 14744919 TI - Can misalignments in typical infants be used as a model for infantile esotropia? AB - PURPOSE: To investigate the nature of early ocular misalignments in human infants to determine whether they can provide insight into the etiology of esotropia and, in particular, to examine the correlates of misalignments. METHODS: A remote haploscopic photorefraction system was used to measure accommodation and vergence in 146 infants between 0 and 12 months of age. Infants underwent photorefraction immediately after watching a target moving between two of five viewing distances (25, 33, 50, 100, and 200 cm). In some instances, infants were tested in two conditions: both eyes open and one eye occluded. The resultant data were screened for instances of large misalignments. Data were assessed to determine whether accommodative, retinal disparity, or other cues were associated with the occurrence of misalignments. RESULTS: The results showed that there was no correlation between accommodative behavior and misalignments. Infants were more likely to show misalignments when retinal disparity cues were removed through occlusion. They were also more likely to show misalignments immediately after the target moved from a near to a far position in comparison to far-to-near target movement. DISCUSSION: The data suggest that the prevalence of misalignments in infants of 2 to 3 months of age is decreased by the addition of retinal disparity cues to the stimulus. In addition, target movement away from the infant increases the prevalence of misalignments. These data are compatible with the notion that misalignment are caused by poor sensitivity to targets moving away from the infant and support the theory that some forms of strabismus could be related to failure in a system that is sensitive to the direction of motion. PMID- 14744920 TI - Cardiovascular risk factors and sequelae in hypertensive children identified by referral versus school-based screening. AB - To determine whether systematic differences exist between hypertensive children referred for evaluation by primary care providers and children identified through community-based screening, cardiovascular risk factors and surrogate markers of hypertensive injury were compared based on subject source (referral versus screening). Children referred to a hypertension clinic for persistently elevated blood pressure were compared with children identified as hypertensive during school screening of 5102 students in Houston public schools. M-mode echocardiography of the left ventricle was performed and subsequently reviewed by 2 independent sonographers blinded to identifying subject information. Subsets of subjects also underwent carotid artery ultrasound for measurement of intimal medial thickness, overnight urine collections for microalbuminuria, and fasting serum cholesterol, triglycerides, and glucose. Ninety-seven total subjects (54 screening and 43 referral) met inclusion criteria and had technically adequate echocardiography performed. The prevalence of left ventricular hypertrophy (LVH) was 37%. Referral subjects demonstrated significantly greater left ventricular mass index (38.8 versus 34.2 g/m(2.7); P<0.01) and a higher prevalence of LVH (49% versus 28%; P<0.05). Among subjects who underwent carotid ultrasound (n=75), carotid intimal-medial thickness was significantly higher in referral subjects (0.61 versus 0.57, P<0.05). When controlling for BMI z score, which was significantly higher in referral subjects, systematic differences by subject source did not persist. These findings suggest that hypertensive children who are predominantly overweight, independent of the manner in which patients come to medical attention, will manifest evidence of more severe cardiovascular disease assessed by surrogate markers such as left ventricular mass index or carotid artery intimal medial thickness. PMID- 14744921 TI - Effects of COX inhibition on blood pressure and kidney function in ACE inhibitor treated blacks and hispanics. AB - Cyclo-oxygenase (COX) inhibitors attenuate the antihypertensive effects of angiotensin-converting enzyme (ACE) inhibitors and reduce kidney function. The study tests the hypothesis that these two classes of drugs have similar effects on glomerular filtration rate (GFR) and 24-hour blood pressure. The primary endpoint was change in 24-hour systolic blood pressure. Using a randomized crossover design, 25 black and Hispanic hypertensive participants (mean age 58+/ 3 years) with osteoarthritis were studied. All participants received an ACE inhibitor at baseline. Once systolic blood pressure was <140 mm Hg, either celecoxib 200 mg/d or diclofenac 75 mg twice daily for 4 weeks was started. After measurements were obtained, all participants underwent a 2-week washout period and crossed over to the other drug for 4 weeks. A significant difference in mean 24-hour systolic blood pressure was noted between groups at 4 weeks (+4.1+/-1.1 mm Hg diclofenac versus +0.6+/-0.6 mm Hg celecoxib; P=0.01). However, because celecoxib has duration of action shorter than 24 hours, we compared ambulatory values at celecoxib trough and peak activities. At peak, no difference in systolic blood pressure was noted between agents (+3.6+/-0.04 mm Hg diclofenac versus +4.2+/-1.9 mm Hg celecoxib; P=0.67). GFR was also differentially affected at 24 hours (-9.9+/-2.4 mL/min diclofenac versus -0.4+/-1.2 mL/min celecoxib; P=0.01). We conclude that diclofenac and celecoxib increase systolic blood pressure at peak levels; however, these agents differ in their 24-hour effects. Differences observed in blood pressure response between COX inhibitors may not be related in their sensitivity but rather their dosing frequency. PMID- 14744922 TI - Childhood blood pressure as a predictor of arterial stiffness in young adults: the bogalusa heart study. AB - Increased arterial stiffness is an independent predictor of cardiovascular disease and mortality in middle-aged and older adults. However, limited data are available regarding the relationship of arterial stiffness in young adults with risk factors measured in childhood, adulthood, or as a cumulative burden from childhood to adulthood. This aspect was examined in a sample of 835 black and white young adults (72% whites, 44% men) aged 24 to 44 years who had at least 4 measurements of traditional risk factors over an average follow-up period of 26.5 years since childhood. Brachial-ankle pulse wave velocity (baPWV) measured by a simple automatic oscillometric technique was used as an index of arterial stiffness. The cumulative burden of risk factors since childhood was measured as area under the curve divided by follow-up years. In young adults, the baPWV was higher in males versus females (P<0.001) and blacks versus whites (P<0.001). In multiple regression analyses, independent predictors of baPWV in young adults were systolic blood pressure in childhood; systolic blood pressure, high-density lipoprotein cholesterol, triglycerides, and smoking in adulthood; and cumulative burden of systolic blood pressure and triglycerides and duration of smoking years from childhood. Thus, systolic blood pressure beginning in childhood is a consistent predictor of arterial stiffness in free-living, asymptomatic young adults. These findings underscore the importance of childhood blood pressure in the evolution of arterial stiffness and the need for beginning preventive cardiology early in life. PMID- 14744923 TI - L-arginine depletion in preeclampsia orients nitric oxide synthase toward oxidant species. AB - Less nitric oxide (NO)-dependent vasodilation and excess formation of reactive oxygen species could explain poor placenta perfusion in preeclampsia, but the pathways involved are unknown. We tested the hypothesis that reduced NO activity and increased oxidative stress in preeclamptic placenta is related to a low bioavailability of l-arginine. Placental endothelial NO synthase (ecNOS) expression (by immunoperoxidase) and activity (by diaphorase and [(3)H]L citrulline formation) were comparable in normotensive pregnancy and in preeclampsia, whereas nitrotyrosine staining, a marker of peroxynitrite, was stronger in preeclamptic villi, confirming previously reported data. Oxidative tissue damage was documented in preeclamptic villi by strong 4-hydroxynonenal lysine staining (by immunoperoxidase), which closely colocalized with nitrotyrosine. Concentration of the NO precursor l-arginine (by HPLC) in umbilical blood and in villous tissue was lower in preeclampsia than in normotensive pregnancy. This was not caused by a defective l-arginine transport, because gene expression of the CAT-1, 4F2hc, and LAT-1 cationic amino acid transporters (by real-time reverse-transcription polymerase chain reaction [RT PCR]) was normal. Instead, gene expression (by real-time RT-PCR) and protein tissue content (by immunoperoxidase and Western blot) of arginase II-the enzyme that degrades arginine to ornithine-were higher in preeclamptic villi than in normotensive pregnancy. These results provide a biochemical explanation for defective NO activity and increased oxidative stress in preeclamptic placenta. In normal placenta, adequate concentration of l-arginine orients ecNOS toward NO. In preeclampsia, a lower than normal l-arginine concentration caused by arginase II overexpression redirects ecNOS toward peroxynitrite. PMID- 14744924 TI - Angiotensin II stimulates calcium and nitric oxide release from Macula densa cells through AT1 receptors. AB - A fluorescent nitric oxide (NO) indicator, 4,5-diaminofluorescein diacetate, and the calcium indicator, indo-1, with 488 nm and 364 nm UV confocal laser scanning microscopy were used to detect NO and calcium concentration in rabbit macula densa (MD) cells challenged by angiotensin II (Ang II). Glomeruli with attached thick ascending limbs with the MD plaque were isolated and perfused. Ang II concentration from 10(-9) to 10(-5) progressively increased MD cell calcium and NO to peak values at 10(-6) and 10(-7), respectively. Ang II (10(-6) M) caused the cytosolic calcium concentration ([Ca(2+)](i)) to increase by 125.8+/-16.3 nM (n=17) from the bath and by 52.3+/-11.5 nM (n=18) from the lumen. AT(1) antagonist CV-11974 (10(-6) M) blocked the Ang II-induced calcium responses from bath and lumen, but AT(2) antagonist PD-123319 (10(-6) M) did not. AT(2) agonist CGP-42112A (10(-6) M) did not affect [Ca(2+)](i) in MD cells from either side. Ang II (10(-6) M) increased the NO production by 16%+/-3.4% (n=26) from the bath and by 18%+/-3.1% (n=24) from the lumen. CV-11974 (10(-6) M) blocked the NO responses from both sides, but PD-123319 (10(-6) M) did not on either side. CGP 42112A (10(-6) M) had no effect on NO in MD cells. In calcium-free experiments there was no difference from the result in normal calcium solutions. In conclusion, we found that Ang II increased [Ca(2+)](i) and stimulated NO production in MD cells from the basolateral and luminal sides through AT(1) receptors. PMID- 14744925 TI - Functional variant in the (alpha)2B adrenoceptor gene, a positional candidate on chromosome 2, associates with hypertension. AB - In a genome-wide scan in Scandinavians, we found suggestive linkage between early onset primary hypertension and a region on chromosome 2. The alpha(2B) adrenoceptor gene, a candidate gene within this region, harbors a functional insertion/deletion (I/D) polymorphism of three glutamate residues. The aim of this study was to investigate if the DD genotype is associated with hypertension in Swedes. We performed an association study between the I/D polymorphism of the alpha(2B)-adrenoceptor and hypertension in the Skaraborg population. The material consists of all known patients with primary hypertension in Skara (n=772 nondiabetic subjects; n=171 normoalbuminuric type 2 diabetic subjects) and 817 population control subjects. We first compared genotype frequencies between patients with early-onset hypertension (aged 50 years or younger at onset) and subjects with normotension (blood pressure <120/80 mm Hg). Thereafter, the polymorphism was tested for association with hypertension at the population level. When comparing patients with early-onset hypertension and normotensive subjects, the DD versus II genotype was associated with early-onset hypertension when diabetic subjects were excluded from the analysis (OR=2.0; 95% CI=1.2 to 3.5) or when they were not excluded (OR=1.8; 95% CI=1.0 to 3.1). At the population level, the DD versus II genotype was weakly associated with nondiabetic hypertension (OR=1.4; 95% CI=1.0 to 1.8). Our data suggest that carriers of the DD versus II genotype of the alpha(2B)-adrenoceptor are at increased risk for hypertension. The genotypic effect is most evident when comparing groups corresponding to the upper and lower tails of the blood pressure distribution in the population; however, in nondiabetic hypertensive subjects it is weakly detectable even at the population level. PMID- 14744926 TI - Effects of heart rate changes on arterial distensibility in humans. PMID- 14744928 TI - Sympathetic responses to stress and rilmenidine in 2K1C rabbits: evidence of enhanced nonvascular effector mechanism. AB - We determined whether the sympathetic excitatory responses to environmental stressors and the sympathoinhibitory responses to rilmenidine are altered by renovascular hypertension. Rabbits were made hypertensive with a clip on the right renal artery, and a left renal nerve recording electrode was implanted. After 3 or 6 weeks, the animals were given air-jet stress and loud noise stress before and after intravenous rilmenidine. Three and 6 weeks after renal clipping, mean arterial pressure was 28% and 36% greater than preclip values. Air-jet stress elicited a marked increase in renal sympathetic nerve activity, mean arterial pressure, and heart rate. Renal sympathetic nerve activity responses were much greater in hypertensive rabbits, but the pressor responses were similar to those observed in normotensive animals. Acute administration of rilmenidine decreased blood pressure more in hypertensive animals but with a much lesser inhibition of sympathetic activity. Rilmenidine markedly reduced increased sympathetic activity during air-jet stress in 3-week clipped rabbits but to a lesser extent in the other groups. These studies show that while sympathetic responses to stress were markedly enhanced in renal clip hypertensive rabbits, they did not result in greater pressor responses, thus suggesting that vascular neuroeffector mechanisms were not altered. By contrast, the increased effects of rilmenidine suggest a much greater contribution to the hypertension by the sympathetic nervous system, but one that is caused by an enhanced "nonvascular" neuroeffector mechanism. As such, sympathoinhibitory agents such as rilmenidine are very suitable and very effective agents for the treatment of renovascular hypertension. PMID- 14744929 TI - Factors associated with hypertension awareness, treatment, and control in a representative sample of the chinese population. AB - We studied factors associated with awareness, treatment, and control of hypertension (systolic/diastolic blood pressure measurements >/=140/90 mm Hg, respectively, and/or current drug treatment for hypertension) in a representative sample of the Chinese population (n=15 838). Awareness, treatment, and control were defined by self-report of a hypertension diagnosis, self-report of current antihypertensive medication use, and a systolic and diastolic blood pressure <140/90 mm Hg, respectively. Higher awareness (OR; 95% CI) was noted for persons who were married (1.43; 1.09, 1.88) and had their blood pressure measured in 1 year (47.4; 31.7, 70.4) or 1 to 5 years (11.4; 7.09, 18.2) preceding their study visit; lower awareness was found among men (0.62; 0.52, 0.74) and current smokers (0.79; 0.63, 0.99). Among those aware of their hypertension, treatment was more common (OR; 95% CI) at higher income (1.24 [0.87, 1.75], 1.46 [1.01, 2.12] and 1.58 [1.06, 2.37] for increasing versus the lowest quartile of income) and for participants who had their blood pressure measured during the preceding year (3.87; 1.89, 7.93) and was less common among men (0.76; 0.59, 0.98). Controlled hypertension was more common (OR; 95% CI) among persons whose blood pressure had been measured 1 year (4.93; 1.51, 16.1) and 1 to 5 years (14.8; 3.63, 60.5) prior to their study visit and, among persons aware of their hypertension diagnosis, those who undertook lifestyle modification (1.59; 1.11, 2.27). These data identify potential methods for improving blood pressure control in China through the identification, follow-up, and lifestyle modification of persons with high blood pressure. PMID- 14744930 TI - PPAR(gamma) agonist rosiglitazone improves vascular function and lowers blood pressure in hypertensive transgenic mice. AB - The peroxisome proliferator activated receptor (PPARgamma) agonist rosiglitazone has been reported to yield cardiovascular benefits in patients by a mechanism that is not completely understood. We tested whether oral rosiglitazone (25 mg/kg per day, 21 days) treatment improves blood pressure and vascular function in a transgenic mouse expressing both human renin and human angiotensinogen transgenes (R(+)A(+)). Rosiglitazone decreased systolic (138+/-5 versus 128+/-5 mm Hg) and mean blood pressure (145+/-5 versus 126+/-7 mm Hg) of R(+)A(+) mice as measured by tail-cuff and indwelling carotid catheters, respectively. Relaxation of carotid arteries to acetylcholine and authentic nitric oxide, but not papaverine, was impaired in R(+)A(+) mice when compared with littermate controls (RA(-)). There were no effects of rosiglitazone on RA(-) mice; however, relaxation to acetylcholine (49+/-10 versus 82+/-9% at 100 micromol/L) and nitric oxide (51+/ 11 versus 72+/-6% at 10 micromol/L) was significantly improved in treated R(+)A(+) mice. Rosiglitazone treatment of R(+)A(+) mice did not alter the expression of genes, including endothelial nitric oxide synthase (eNOS), angiotensin 1 receptors, and preproendothelin-1, nor did it alter the levels of eNOS or soluble guanylyl cyclase protein. In separate studies, carotid arteries from R(+)A(+) and RA(-) mice relaxed in a concentration-dependent manner to rosiglitazone, suggesting possible PPARgamma-independent effects in the vasculature. This response was not inhibited with the nitric oxide synthase inhibitor N(omega)-nitro-l-arginine methyl ester (200 micromol/L) or the PPARgamma antagonist bisphenol A diglycidyl ether; 4,4'-isopropylidenediphenol diglycidyl ether (100 micromol/L). These data suggest that in addition to potential genomic regulation caused by PPARgamma activation, the direct effect of rosiglitazone in blood vessels may contribute to the improved blood pressure and vessel function. PMID- 14744931 TI - Normative values for circadian and ultradian cardiovascular rhythms in childhood. AB - To assess the prevalence and characteristics of physiological circadian (24-hour) and ultradian (12-, 8-, and 6-hour) rhythms of mean arterial blood pressure (BP) and heart rate (HR), we analyzed 24-hour ambulatory BP profiles from 938 healthy school children aged 5 to 18 years. Cosine harmonics were fitted by Fourier analysis, and an amplitude and acrophase (time of peak) were calculated for each rhythm. Ninety percent of children displayed circadian rhythmicity of BP, independent of age, whereas circadian HR rhythmicity decreased with puberty from 96% to 87% (P<0.0001). Puberty had marked effects on the prevalence of ultradian rhythmicity: 12- and 6-hour rhythms increased for BP (27% to 47%, P<0.0001; 18% to 25%, P=0.01) and HR (36% to 47%, 17% to 31%, both P=0.001), whereas 8-hour BP rhythms decreased (34% to 23%, P=0.002). Median amplitudes were 10.1, 5.9, 5.6, and 5.2 mm Hg for the 24-, 12-, 8-, and 6-hour BP rhythms, respectively, and 13.4, 7.7, 6.8, and 6.4 bpm for HR. The acrophase occurred at approximately 14:00 hours, 8:00 hours, 5:30 hours, and 2:00 hours (military time) for the four BP rhythms, and at 13:30 hours, 08:30 hours, 01:50 hours, and 02:00 hours for HR. For the combined curve, the peak-trough difference was 25.9 mm Hg and 35 bpm for BP and HR, respectively, with the peaks occurring at 13:50 hours and 13:10 hours. There was marked association between BP and HR rhythms, both for prevalence (P<0.0001 for coupling of BP and HR rhythms of the same period length) and timing, with a median time lag of BP after HR acrophase of only 21, 16, 13, and 5 minutes for the four rhythms, respectively. PMID- 14744932 TI - Diastolic and pulse pressure: the old and the new? PMID- 14744933 TI - Suppression of mitochondrial respiration through recruitment of p160 myb binding protein to PGC-1alpha: modulation by p38 MAPK. AB - The transcriptional coactivator PPAR gamma coactivator 1 alpha (PGC-1alpha) is a key regulator of metabolic processes such as mitochondrial biogenesis and respiration in muscle and gluconeogenesis in liver. Reduced levels of PGC-1alpha in humans have been associated with type II diabetes. PGC-1alpha contains a negative regulatory domain that attenuates its transcriptional activity. This negative regulation is removed by phosphorylation of PGC-1alpha by p38 MAPK, an important kinase downstream of cytokine signaling in muscle and beta-adrenergic signaling in brown fat. We describe here the identification of p160 myb binding protein (p160MBP) as a repressor of PGC-1alpha. The binding and repression of PGC 1alpha by p160MBP is disrupted by p38 MAPK phosphorylation of PGC-1alpha. Adenoviral expression of p160MBP in myoblasts strongly reduces PGC-1alpha's ability to stimulate mitochondrial respiration and the expression of the genes of the electron transport system. This repression does not require removal of PGC 1alpha from chromatin, suggesting that p160MBP is or recruits a direct transcriptional suppressor. Overall, these data indicate that p160MBP is a powerful negative regulator of PGC-1alpha function and provide a molecular mechanism for the activation of PGC-1alpha by p38 MAPK. The discovery of p160MBP as a PGC-1alpha regulator has important implications for the understanding of energy balance and diabetes. PMID- 14744934 TI - Ovca1 regulates cell proliferation, embryonic development, and tumorigenesis. AB - Loss of OVCA1/DPH2L1 correlates with ovarian and breast cancer. To study its in vivo role, we generated Ovca1 mutant alleles in mice. Ovca1 heterozygotes spontaneously develop cancer. Ovca1 mutant mice die during embryonic development and at birth with developmental delay and defects in multiple organ systems. Cell proliferation defects were observed in Ovca1 mutant mouse embryonic fibroblasts (MEFs). p53 deficiency can rescue these Ovca1 mutant MEF proliferation defects and partially rescue Ovca1 mutant embryonic phenotypes. Furthermore, Ovca1; p53 double heterozygotes developed tumors quicker than p53 heterozygotes and with an increased carcinoma incidence. Multiple tumor burden in Ovca1 heterozygotes that were also p53 deficient was significantly higher than in p53 homozygous mutants. These in vivo findings demonstrate that Ovca1 is a tumor suppressor that can modify p53-induced tumorigenesis and suggest that it acts as a positive regulator for cell cycle progression. The close linkage of OVCA1 and p53 on human Chromosome 17 suggests that coordinated loss may be an important mechanism for the evolution of ovarian, breast, and other tumor phenotypes. PMID- 14744935 TI - Endoplasmic reticulum stress induces p53 cytoplasmic localization and prevents p53-dependent apoptosis by a pathway involving glycogen synthase kinase-3beta. AB - The tumor suppressor p53, a sensor of multiple forms of cellular stress, is regulated by post-translational mechanisms to induce cell-cycle arrest, senescence, or apoptosis. We demonstrate that endoplasmic reticulum (ER) stress inhibits p53-mediated apoptosis. The mechanism of inhibition involves the increased cytoplasmic localization of p53 due to phosphorylation at serine 315 and serine 376, which is mediated by glycogen synthase kinase-3 beta (GSK-3beta). ER stress induces GSK-3beta binding to p53 in the nucleus and enhances the cytoplasmic localization of the tumor suppressor. Inhibition of apoptosis caused by ER stress requires GSK-3beta and does not occur in cells expressing p53 with mutation(s) of serine 315 and/or serine 376 to alanine(s). As a result of the increased cytoplasmic localization, ER stress prevents p53 stabilization and p53 mediated apoptosis upon DNA damage. It is concluded that inactivation of p53 is a protective mechanism utilized by cells to adapt to ER stress. PMID- 14744936 TI - Differential regulation of the human CYP3A4 promoter in transgenic mice and rats. AB - Previously we described a transgenic mouse model [FVB/NTg(CYP3A4-luc)Xen] using a reporter construct consisting of 13 kilobases of the human CYP3A4 promoter driving the firefly luciferase gene in the inbred FVB/N mouse strain. Here we report regulation of the same CYP3A4-luc reporter gene in a transgenic outbred mouse strain (CD-1) and in a transgenic rat (Sprague-Dawley). Basal reporter expression and responses to several xenobiotics in the transgenic CD-1 mice [CD 1/Crl-Tg(CYP3A4-luc)Xen] were similar to those in the transgenic FVB/N mice. In both mouse backgrounds, the basal levels of the reporter were higher in male compared with female, and in the FVB/N strain there was greater induction for all drugs in male compared with female; however, in the CD-1 background this gender difference for induction was not obvious. In contrast with transgenic mice, transgenic rats [SD/Tac-Tg(CYP3A4-luc)Xen] expressed the luciferase reporter at higher basal levels in female compared with male rats. Responses to some compounds were much greater in rats than in mice, and the kinetics of induction was different with peak induction occurring later in the rat compared with the mouse. Our results suggest that the human CYP3A4 promoter is regulated differently in transgenic mice and rats in some aspects. PMID- 14744937 TI - Effect of recombinant interleukin-2 pretreatment on oral and intravenous digoxin pharmacokinetics and P-glycoprotein activity in mice. AB - P-glycoprotein (P-gp) is an ATP-dependent efflux membrane transporter involved in many drug pharmacokinetics in humans. Decreasing its expression could enhance the bioavailability of substrates as digoxin. We have recently found that human recombinant interleukin-2 (rIL2) in vivo decreases P-gp expression in intestine and brain of mice and modifies oral digoxin pharmacokinetics. The aim of the study was to evaluate the involvement of bioavailability in the rIL2 pretreatment effect on digoxin pharmacokinetics by comparing oral and i.v. digoxin pharmacokinetics before and after rIL2 pretreatment (10 microg/kg). We also tried to show the possible effect of a low rIL2 dose (1 microg/kg) pretreatment on oral digoxin pharmacokinetics. First, adult Swiss mice received a single oral or i.v. dose of digoxin (0.03 mg/kg). Two weeks later, the same animals were treated by rIL2 i.p. twice a day (10 microg/kg) for 4 days and received digoxin again at day 5. As well, another group received oral digoxin (0.03 mg/kg) with a 1 microg/kg rIL2 pretreatment. Blood was collected after digoxin administration with and without rIL2 pretreatment. Digoxin pharmacokinetics were described by a one compartment model. The 10 microg/kg rIL2 pretreatment did not modify i.v. digoxin pharmacokinetics, whereas oral digoxin pharmacokinetics were significantly modified by the 10 microg/kg rIL2 pretreatment and not by the 1 microg/kg rIL2 pretreatment. The decrease of P-gp activity, caused by rIL2 (10 microg/kg), increased digoxin bioavailability. An increase in exposure and intracellular level of drugs is expected from rIL2 pretreatment. PMID- 14744938 TI - Development of an in vivo preclinical screen model to estimate absorption and first-pass hepatic extraction of xenobiotics. II. Use of ketoconazole to identify P-glycoprotein/CYP3A-limited bioavailability in the monkey. AB - The effect of P-glycoprotein (Pgp) and/or CYP3A on the disposition of xenobiotics has been extensively investigated and is often of interest during drug discovery lead optimization. We have previously described a monkey pharmacokinetic screen to rapidly estimate absorption and first-pass extraction. In the present work, this monkey screen has been expanded to include an assessment of Pgp/CYP3A effects on absorption and first-pass extraction, using ketoconazole as a prototypic dual Pgp/CYP3A inhibitor. To generate a ketoconazole dosing regimen, the pharmacokinetics of ketoconazole were first determined in the monkey and were found to be consistent with that previously described in the rat, dog, and human. Dose-ranging experiments demonstrated that a single 10-mg/kg intraduodenal ketoconazole dose would provide an appropriate exposure; this dose was used throughout subsequent interaction experiments. Next, erythromycin and propranolol were explored as positive and negative control substrates for Pgp/CYP3A interactions, respectively. As anticipated, ketoconazole produced no change in the absorption or first-pass extraction of propranolol but resulted in a substantial increase in absorption and decrease in first-pass extraction of erythromycin. Finally, this ketoconazole-based monkey screen was deployed in a drug discovery setting, and examples of such use are presented. These experiments have allowed a more complete characterization of ketoconazole as a prototypic dual Pgp/CYP3A inhibitor and its use as a tool in a preclinical setting and further demonstrate the use of the monkey to investigate the role of Pgp/CYP3A in limiting the oral bioavailability of new drug candidates. PMID- 14744939 TI - Identification of a hydroxylamine glucuronide metabolite of an oral hypoglycemic agent. AB - Glucuronides of piperazine hydroxylamines are rarely reported in the literature, and even more rarely are their structures unambiguously identified. One major metabolite was detected by liquid chromatography/mass spectrometry-radioactivity in urine from monkeys treated with the aryl piperazine oral hypoglycemic agent 9 [(1S,2R)-2-fluoro-1-methylpropyl]-2-methoxy-6-(1-piperazinyl) purine hydrochloride (1). The mass spectrum of this metabolite indicated that it was both monooxygenated and glucuronidated on the piperazine ring. Possible structures included the N- or O-glucuronic acid conjugates of a carbinolamine, hydroxylamine, or N-oxide. Treatment with beta-glucuronidase gave a monooxygenated derivative of the parent compound. 1H NMR analysis of either the glucuronic acid conjugate or the monooxygenated product provided insufficient evidence to unambiguously determine their structures. Incubation of 1 with pig liver microsomes resulted in formation of the same monooxygenated derivative derived from beta-glucuronidase treatment of the glucuronide metabolite. This in vitro system was used to generate sufficient material for analysis by 13C NMR, and the metabolite was identified as a hydroxylamine derivative 2. Incubation of the hydroxylamine with monkey liver microsomes and uridine diphospho-5' glucuronic acid gave the same glucuronic acid conjugate as that observed in monkey urine. 13C NMR analysis of this biosynthetic product led to its unequivocal structure assignment as the O-glucuronic acid conjugate of the hydroxylamine 3. PMID- 14744940 TI - Differential enantioselectivity and product-dependent activation and inhibition in metabolism of verapamil by human CYP3As. AB - In vitro studies of enantioselective metabolism of R-(+)- and S-(-)verapamil (VER) were conducted using human cDNA-expressed CYP3A isoforms, CYP3A4, CYP3A5, and CYP3A7. N-dealkylated products nor-VER [2,8-bis-(3,4-dimethoxyphenyl)-2 isopropyl-6-azaoctanitrile] and D617 [2-(3,4-dimethoxyphenyl)-5-methylamino-2 isopropylvaleronitrile] were the major metabolites for all CYP3A isoforms regardless of enantiomer. Enantioselectivity of CYP3A4 and CYP3A7 was most similar among the three isoforms. This coincides with the degree of homology of amino acids at the active sites and in the total amino acid sequences of the enzymes. Biphasic substrate inhibition was observed for the formation of nor-VER and D617, whereas simple biphasic kinetics were observed for the formation of O demethylated products for both enantiomers with CYP3A4. The biphasic substrate inhibition was observed only for nor-VER, and simple biphasic kinetics were observed for D617 and O-demethylated products for both enantiomers with CYP3A5. However, with CYP3A7, D617 and O-demethylated products showed typical Michaelis Menten kinetics, and only nor-VER displayed substrate (monophasic) inhibition. When metabolic rates of VER were determined in the presence of three different effectors, midazolam, testosterone, and nifedipine, activation, inhibition, or activation and inhibition of VER metabolism was observed depending on the enantiomers, metabolites, effectors, and cytochrome P450 isoforms. Addition of anti-CYP3A4 antibody inhibited formation of all metabolites for both CYP3A4 and CYP3A5. The atypical phenomena (biphasic substrate inhibition, activation, and inhibition depending on product formation) of VER kinetics could be adequately explained by introducing the concept of steric interaction into a two binding site model. PMID- 14744941 TI - Nonlinear oral pharmacokinetics of the alpha-antagonist 4-amino-5-(4 fluorophenyl)-6,7-dimethoxy-2-[4-(morpholinocarbonyl)-perhydro-1,4-diazepin-1 yl]quinoline in humans: use of preclinical data to rationalize clinical observations. AB - 4-amino-5-(4-fluorophenyl)-6,7-dimethoxy-2-[4-(morpholinocarbonyl)-perhydro-1,4 diazepin-1-yl]quinoline (UK-294,315) is an antagonist of the human alpha1 adrenoceptor and exhibits nonlinear oral pharmacokinetics in humans. Superproportional increases in Cmax occur (220-fold, over a 1- to 50-mg dose range), area under the curve increases linearly, but time to maximum concentration decreases with dose, suggesting variation in rate but not extent of absorption. Oral absorption in humans is extensive, with only 14% of an orally administered (20 mg) radiolabeled dose excreted unchanged in the feces. In rats and dogs, UK-294,315 is partially eliminated as unchanged drug in feces (29 and 14% of an intravenous dose, respectively). Oral bioavailability is low in rats (11%) and high in dogs (71%), in keeping with systemic clearance. Fecal elimination of unchanged drug was 60% after oral administration to rats, indicating incomplete absorption in this species, whereas absorption in dogs is complete. UK-294,315 is a P-glycoprotein (P-gp) substrate (Km, 15 microM) exhibiting polarized flux in Caco-2 cell monolayers, saturable across a concentration range of 5 to 200 microM. Furthermore, the observations in vitro occurred at similar concentrations to those estimated in the gut lumen in clinical trials (dose range, 1-100 mg). It is considered that P-gp acts as a saturable absorption barrier to UK-294,315, slowing the rate of absorption at low doses, and is responsible for the observed nonlinearity in oral disposition in humans. Rat and dog pharmacokinetic studies offered limited insight into the process(es) driving nonlinear pharmacokinetics in humans. Our current understanding of the functional effects of P-gp in the human intestine, in combination with in vitro studies at clinically relevant concentrations, has helped rationalize the clinical data for UK-294,315. PMID- 14744942 TI - Biotransformation of fucoxanthinol into amarouciaxanthin A in mice and HepG2 cells: formation and cytotoxicity of fucoxanthin metabolites. AB - Fucoxanthin, a major carotenoid in edible brown algae, potentially inhibits the proliferation of human prostate cancer cells via apoptosis induction. However, it has been postulated that dietary fucoxanthin is hydrolyzed into fucoxanthinol in the gastrointestinal tract before absorption in the intestine. In the present study, we investigated the further biotransformation of orally administered fucoxanthin and estimated the cytotoxicity of fucoxanthin metabolites on PC-3 human prostate cancer cells. After the oral administration of fucoxanthin in mice, two metabolites, fucoxanthinol and an unknown metabolite, were found in the plasma and liver. The unknown metabolite was isolated from the incubation mixture of fucoxanthinol and mouse liver preparation (10,000 g supernatant of homogenates), and a series of instrumental analyses identified it as amarouciaxanthin A [(3S,5R,6'S)-3,5,6'-trihydroxy-6,7-didehydro-5,6,7',8' tetrahydro-beta,epsilon-carotene-3',8'-dione]. The conversion of fucoxanthinol into amarouciaxanthin A was predominantly shown in liver microsomes. This dehydrogenation/isomerization of the 5,6-epoxy-3-hydroxy-5,6-dihydro-beta end group of fucoxanthinol into the 6'-hydroxy-3'-oxo-epsilon end group of amarouciaxanthin A required NAD(P)+ as a cofactor, and the optimal pH for the conversion was 9.5 to 10.0. Fucoxanthinol supplemented to culture medium via HepG2 cells was also converted into amarouciaxanthin A. The 50% inhibitory concentrations on the proliferation of PC-3 human prostate cancer cells were 3.0, 2.0, and 4.6 microM for fucoxanthin, fucoxanthinol, and amarouciaxanthin A, respectively. To our knowledge, this is the first report on the enzymatic dehydrogenation of a 3-hydroxyl end group of xanthophylls in mammals. PMID- 14744943 TI - Stereoselective pharmacokinetics of fluoxetine and norfluoxetine enantiomers in pregnant sheep. AB - We examined the stereoselective disposition of fluoxetine (FX) and its metabolite norfluoxetine (NFX) in five pregnant sheep. Racemic FX was administered i.v. to the ewe (50 mg) and the fetus (10 mg) on separate occasions. Maternal and fetal blood, maternal urine, and fetal amniotic and tracheal fluid samples were collected for 72 h. FX and NFX isomers were quantified by gas chromatography-mass spectrometry. They rapidly crossed the placenta [maternal to fetal area under the plasma concentration versus time curve (AUC) ratios 0.59 and 0.65, respectively]. There was significant FX stereoselectivity with S/R FX AUC ratios averaging 1.65 +/- 0.33 and 1.73 +/- 0.29 in ewe and fetus, respectively, after maternal dosing. The maternal clearance and volume of distribution were also higher for (R) fluoxetine than for (S)-fluoxetine. FX, NFX, and their glucuronides were present in maternal urine but accounted for only 3.4% of maternal drug elimination. In contrast, NFX was not detected in the fetus after fetal FX administration, which is consistent with the absence of measurable fetal nonplacental clearance of the drug and the lack of NFX formation in fetal hepatic microsomal incubations. There was also no fetal production of FX and NFX glucuronides in vivo. Both FX and NFX were extensively and stereoselectively bound in maternal and fetal plasma, with the free fraction S/R FX ratio averaging 0.46 +/- 0.06 and 0.58 +/- 0.10 in ewe and fetus, respectively. Thus, FX exhibits extensive stereoselective disposition, which is likely due to differential plasma protein binding of the FX isomers, and there is no detectable fetal formation of NFX, FX, and NFX glucuronides. PMID- 14744944 TI - Effect of total parenteral nutrition and choline on hepatic flavin-containing and cytochrome P-450 monooxygenase activity in rats. AB - Total parenteral nutrition provides nutrition by infusion into the systemic circulation. Bypassing the intestine and processes associated with absorption can cause additional pathophysiological changes to occur. For example, in rats, normal gut and pancreatic cell function may change, absorptive capacity may be altered, and enzyme functional activity including drug metabolism may be affected. The objective of this study was to examine the effects of a control diet or a diet of total parenteral nutrition in the presence or absence of choline on urinary biomarkers and hepatic microsome functional activity from rats. Selective functional markers of cytochrome P-4502E1 (CYP2E1) and flavin containing monooxygenase (FMO) were examined in vitro. The N-oxygenation of trimethylamine was used as an in vivo selective functional marker for FMO. After the administration of total parenteral nutrition plus choline for 5 days, the urinary excretion of trimethylamine and trimethylamine N-oxide declined approximately 7- and 3-fold, respectively, compared with rats treated with control diet. The concentration of urinary biogenic amines was also significantly affected by total parenteral nutrition. Compared with control animals, rats administered total parenteral nutrition plus choline for 5 days showed a decrease of approximately 5- and 2-fold in urinary dopamine and norepinephrine concentration, respectively. To examine a molecular basis for the influence of total parenteral nutrition +/- choline on monooxygenase regulation, hepatic microsomal activity of the FMO and CYP2E1 was examined. Compared with animals treated with a control diet, total parenteral nutrition plus choline in rats caused a 3-fold increase in hepatic microsomal FMO and a 2-fold increase in hepatic cytochrome CYP2E1 functional activity, respectively. Although the data did not reach statistical significance, selective immunoblot studies using hepatic microsomes from rats treated with total parenteral nutrition + choline showed that compared with controls, FMO1 protein was decreased 1.4-fold and FMO3 increased 1.3-fold, respectively. In hepatic microsomes from rats treated with total parenteral nutrition + choline, compared with control animals, FMO4 immunoreactivity was increased 1.6-fold. The data suggest that total parenteral nutrition has a detectable effect on modulating rat FMO3, FMO4, and CYP2E1 monooxygenase functional activity. The clinical relevance of these results is unknown but may be of significance for individuals receiving total parenteral nutrition and those afflicted with trimethylaminuria. PMID- 14744945 TI - An evaluation method for nonlinear local disposition in rat liver and kidney. AB - A two-sampling sites method was developed to separately estimate the nonlinear local disposition in the liver and kidney by sampling blood simultaneously from the hepatic vein and an artery after intravenous administration. Using this method, it was attempted to predict the renal elimination from the systemic and hepatic elimination. Etoposide, a substrate of both P-glycoprotein and CYP3A, was used as a model drug. The blood samples from the hepatic vein and an artery were simultaneously taken from a rat after intravenous administration of etoposide at a dose of 20 or 80 mg/kg. At a dose of 20 mg/kg, the total clearance (CL), hepatic clearance (CLH), and renal clearance (CLR=CL-CLH), which were almost constant, were 2.82 +/- 0.24, 0.742 +/- 0.214, and 2.09 +/- 0.34 l/h/kg, respectively. At a dose of 80 mg/kg, CL and CLR considerably decreased with an increase in plasma concentration, whereas CLH slightly decreased. By means of the two-sampling sites method, we estimated the local drug disposition in the liver and kidney. The present local pharmacokinetic method would be applicable to assess the local disposition of other drugs that are mainly metabolized in these organs. PMID- 14744946 TI - Involvement of human hepatic UGT1A1, UGT2B4, and UGT2B7 in the glucuronidation of carvedilol. AB - Carvedilol ((+/-)-1-carbazol-4-yloxy)-3-[[2-(o-methoxyphenoxy)ethyl]amino]-2 propanol) is metabolized primarily into glucuronide conjugates. In the present study, we identified the human UDP-glucuronosyltransferase (UGT) isoforms involved in the glucuronidation of carvedilol by thin-layer chromatography using microsomes from human liver or insect cells expressing recombinant UGT isoforms. We observed two forms of carvedilol glucuronides, namely G1 and G2, in hepatic microsomes. The glucuronidation of carvedilol was catalyzed by at least three recombinant UGT isoforms: UGT1A1, UGT2B4, and UGT2B7. UGT2B4 formed both G1 and G2, whereas UGT1A1 and UGT2B7 were responsible for the formation of glucuronide G2 and G1, respectively. The enzyme kinetics for carvedilol glucuronidation by UGT1A1, UGT2B4, and UGT2B7 in addition to human liver microsomes were examined by Lineweaver-Burk analysis. The values of Km and Vmax for human liver microsomes were 26.6 microM and 106 pmol/min/mg protein for G1, and 46.0 microM and 44.5 pmol/min/mg protein for G2, respectively. The Km values for UGT1A1, UGT2B4, and UGT2B7 for G1 and G2 (22.1-55.1 microM) were comparable to those of the liver microsomes, whereas the Vmax values were in the range of 3.33 to 7.88 pmol/min/mg protein. The Km and Vmax/Km values for UGT2B4 and UGT2B7 for G1 were similar, whereas UGT2B4 had lower Km and higher Vmax/Km values for G2 compared with those of UGT1A1. These results suggest that G1 formation is catalyzed by UGT2B4 and UGT2B7, whereas G2 is formed by UGT2B4 and UGT1A1. These three hepatic UGT isoforms may have important roles in carvedilol metabolism. PMID- 14744947 TI - Polymorphic expression of CYP1A2 leading to interindividual variability in metabolism of a novel benzodiazepine receptor partial inverse agonist in dogs. AB - 5-(3-methoxyphenyl)-3-(5-methyl-1,2,4-oxadiazol-3-yl)-2-oxo-1,2-dihydro-1,6 naphthyridine (AC-3933) is a novel cognitive enhancer with central benzodiazepine receptor partial inverse agonistic activity. AC-3933 is predominantly metabolized to hydroxylated metabolite [SX-5745; 3-(5-hydroxymethyl-1,2,4-oxadiazol-3-yl)-5 (3-methoxyphenyl)-2-oxo-1,2-dihydro-1,6-naphthyridine] in dog. Initially, we found that there is considerable interindividual variability in AC-3933 hydroxylation in dogs and that dogs could be phenotyped as extensive metabolizer (EM) and poor metabolizer (PM). Then, to clarify the cause of AC-3933 polymorphic hydroxylation in dogs, in vitro studies were carried out using liver microsomes from EM and PM dogs. Our results show that AC-3933 hydroxylation clearance in PM dogs was much lower than that in EM dogs (0.2 versus 10.8-20.5 microl/min/mg, respectively). In addition, AC-3933 hydroxylation was significantly inhibited by alpha-naphthoflavone, a CYP1A inhibitor, and by anti-CYP1A2 antibodies, indicating that CYP1A2 was responsible for the polymorphic hydroxylation of AC 3933 in dogs. Furthermore, immunoblotting results have shown that although CYP1A2 protein was not detected in PM dogs (<0.86 pmol/mg), CYP1A2 content in EM dogs was prominent (6.1-13.0 pmol/mg). These results indicate that AC-3933 polymorphic hydroxylation arises from the polymorphic expression of CYP1A2 in dogs, which might involve genetic polymorphism of the CYP1A2 gene. PMID- 14744949 TI - Prediction of cytochrome P450 3A inhibition by verapamil enantiomers and their metabolites. AB - Verapamil inhibition of CYP3A activity results in many drug-drug interactions with CYP3A substrates, but the mechanism of inhibition is unclear. The present study showed that verapamil enantiomers and their major metabolites [norverapamil and N-desalkylverapamil (D617)] inhibited CYP3A in a time- and concentration dependent manner by using pooled human liver microsomes and the cDNA-expressed CYP3A4 (+b5). The values of the inactivation kinetic parameters kinact and KI obtained with the cDNA-expressed CYP3A4 (+b5) were 0.39 min(-1) and 6.46 microM for R-verapamil, 0.64 min(-1) and 2.97 microM for S-verapamil, 1.12 min(-1) and 5.89 microM for (+/-)-norverapamil, and 0.07 min(-1) and 7.93 microM for D617. Based on the ratio of kinact and KI, the inactivation potency of verapamil enantiomers and their metabolites was in the following order: S-norverapamil>S verapamil>R-norverapamil>R-verapamil>D617. Using dual beam spectrophotometry, we confirmed that metabolic intermediate complex formation with CYP3A was the mechanism of inactivation for all compounds. The in vitro unbound fraction was 0.84 for S-verapamil, 0.68 for R-verapamil, and 0.84 for (+/-)-norverapamil. A mechanism-based pharmacokinetic model predicted that the oral area under the curve (AUC) of a CYP3A substrate that is eliminated completely (fm=1) by the hepatic CYP3A increased 1.6- to 2.2-fold after repeated oral administration of verapamil. For midazolam (fm=0.9), a drug that undergoes extensive intestinal wall metabolism, the predicted increase in oral AUC was 3.2- to 4.5-fold. The predicted results correlate well with the in vivo drug interaction data, suggesting that the model is suitable for predicting drug interactions by mechanism-based inhibitors. PMID- 14744948 TI - The metabolic disposition of aprepitant, a substance P receptor antagonist, in rats and dogs. AB - The absorption, metabolism, and excretion of [14C]aprepitant, a potent and selective human substance P receptor antagonist for the treatment of chemotherapy induced nausea and vomiting, was evaluated in rats and dogs. Aprepitant was metabolized extensively and no parent drug was detected in the urine of either species. The elimination of drug-related radioactivity, after i.v. or p.o. administration of [14C]aprepitant, was mainly via biliary excretion in rats and by way of both biliary and urinary excretion in dogs. Aprepitant was the major component in the plasma at the early time points (up to 8 h), and plasma metabolite profiles of aprepitant were qualitatively similar in rats and dogs. Several oxidative metabolites of aprepitant, derived from N-dealkylation, oxidation, and opening of the morpholine ring, were detected in the plasma. Glucuronidation represented an important pathway in the metabolism and excretion of aprepitant in rats and dogs. An acid-labile glucuronide of [14C]aprepitant accounted for approximately 18% of the oral dose in rat bile. The instability of this glucuronide, coupled with its presence in bile but absence in feces, suggested the potential for enterohepatic circulation of aprepitant via this conjugate. In dogs, the glucuronide of [14C]aprepitant, together with four glucuronides derived from phase I metabolites, were present as major metabolites in the bile, accounting collectively for approximately 14% of the radioactive dose over a 4- to 24-h period after i.v. dosing. Two very polar carboxylic acids, namely, 4-fluoro-alpha-hydroxybenzeneacetic acid and 4-fluoro-alpha oxobenzeneacetic acid, were the predominant drug-related entities in rat and dog urine. PMID- 14744950 TI - Identification of human UDP-glucuronosyltransferase enzyme(s) responsible for the glucuronidation of posaconazole (Noxafil). AB - Posaconazole (Noxafil, SCH 56592), an orally available broad-spectrum triazole antifungal, is currently in phase III clinical studies for treating serious opportunistic fungal infections. The major in vitro metabolite of posaconazole formed by human liver microsomes supplemented with uridine 5'-diphosphate glucuronic acid was a glucuronide of posaconazole (m/z877). Screening of 10 cDNA expressed recombinant human UDP-glucuronosyltransferase (UGT) enzymes showed that only UGT1A4 exhibited catalytic activity with respect to the formation of the glucuronide of posaconazole. The formation of glucuronide by human liver microsomes and UGT1A4 was inhibited by bilirubin, a known inhibitor of UGT1A4. There was a high correlation (r =0.90) between the rate of formation of glucuronide, determined in 10 human liver microsomal samples, and trifluoperazine glucuronidation catalyzed by UGT1A4. These results confirmed that the formation of major posaconazole-glucuronide produced from human liver microsomes was mediated via UGT1A4. PMID- 14744951 TI - Metabolism of 8-prenylnaringenin, a potent phytoestrogen from hops (Humulus lupulus), by human liver microsomes. AB - The female flowers of hops are used throughout the world as a flavoring agent for beer. Recently, there has been increasing interest in the potential estrogenic properties of hop extracts. Among the possible estrogenic compounds in hops, 8 prenylnaringenin is perhaps most significant due to its high in vitro potency exceeding that of other known phytoestrogens. Since data regarding the pharmacokinetic properties of this compound are lacking, we investigated the in vitro metabolism of 8-prenylnaringenin by human liver microsomes. A total of 12 metabolites were identified, and biotransformation occurred on the prenyl group and the flavanone skeleton. The major site of oxidation was on the terminal methyl groups, and of the two possible isomers, the transisomer was more abundant. The double bond on the prenyl group was also oxidized to an epoxide that was opened by intramolecular reaction with the neighboring hydroxyl group. On the flavanone skeleton, the major site of oxidation was at 3'position on the B ring. Other metabolites included oxidation at carbon-3 as well as desaturation of the C ring to produce 8-prenylapigenin. An unusual hydroxy quinone product formed by ipso hydroxylation of the B ring of 8-prenylnaringenin was also detected. This product was probably an intermediate for the B ring cleavage product, 8 prenylchromone. PMID- 14744952 TI - Renal disease, homocysteine, and cardiovascular complications. PMID- 14744953 TI - Ventricular resynchronization: current state of the art. PMID- 14744954 TI - Cardiac resynchronization therapy: current state of the art: cost versus benefit. PMID- 14744955 TI - Cardiac resynchronization therapy: helpful now in selected patients with CHF. PMID- 14744956 TI - Microvasculature in acute myocardial ischemia: part II: evolving concepts in pathophysiology, diagnosis, and treatment. PMID- 14744957 TI - Imaging of intracoronary thrombus by multidetector helical computed tomography angiography. PMID- 14744958 TI - Definition of metabolic syndrome: Report of the National Heart, Lung, and Blood Institute/American Heart Association conference on scientific issues related to definition. PMID- 14744959 TI - Cardiology patient page. New blood tests for detecting heart disease. PMID- 14744960 TI - Diagnosis of hemi-truncus arteriosis by three-dimensional magnetic resonance angiography. PMID- 14744961 TI - Vasodilation during systemic tyramine administration response. PMID- 14744962 TI - Selective serotonin reuptake inhibitors and myocardial infarction. PMID- 14744963 TI - C-reactive protein in patients with acute myocardial infarction. PMID- 14744964 TI - All opinions are not equal. PMID- 14744965 TI - Drug-eluting stents: do we know enough about safety? PMID- 14744967 TI - Value of d-dimer testing in acute aortic dissection. PMID- 14744966 TI - Metabolic syndrome and diabetes mellitus. PMID- 14744969 TI - Hydroxymethyl-glutaryl coenzyme a reductase inhibition limits cytomegalovirus infection in human endothelial cells. AB - BACKGROUND: Statins exert anti-inflammatory effects independently of cholesterol lowering properties. Cytomegalovirus (CMV) infection appears to be implicated in the pathophysiology of atherosclerosis by inducing inflammatory modifications in endothelial cells, especially in immunosuppressed patients. We investigated whether the activity of statins can inhibit replication of CMV in human endothelial cells. METHODS AND RESULTS: Human umbilical vein endothelial cells (HUVECs) were infected with CMV and coincubated with fluvastatin at 0.1 and 0.2 micromol/L. Fluvastatin inhibited (P<0.001) CMV antigen expression, and this effect was dose related (P<0.001). Quantitative polymerase chain reaction showed that CMV DNA concentration was consistently lower in supernatants from fluvastatin-treated cells than in infected controls, and viral particle concentration was up to 30 times lower in 0.2 micromol/L fluvastatin-treated cells than in infected controls (10.5+/-0.9 versus 0.34+/-0.03 per 10(3) pfu/mL, P<0.001). Addition of mevalonate to treated cultures almost completely abolished fluvastatin inhibition of viral growth. Electrophoretic mobility shift assay showed that fluvastatin reduced nuclear factor-kappaB binding activity in CMV infected cells. CONCLUSIONS: HMG-CoA inhibition by fluvastatin restrains CMV replication in HUVECs by inhibiting viral antigen expression, DNA synthesis, and viral particle production, conceivably by involving a reduction of nuclear factor kappaB binding activity. PMID- 14744970 TI - Determinants and prognostic impact of heart failure complicating acute coronary syndromes: observations from the Global Registry of Acute Coronary Events (GRACE). AB - BACKGROUND: Few data are available on the impact of heart failure (HF) across all types of acute coronary syndromes (ACS). METHODS AND RESULTS: The Global Registry of Acute Coronary Events (GRACE) is a prospective study of patients hospitalized with ACS. Data from 16 166 patients were analyzed: 13 707 patients without prior HF or cardiogenic shock at presentation were identified. Of these, 1778 (13%) had an admission diagnosis of HF (Killip class II or III). HF on admission was associated with a marked increase in mortality rates during hospitalization (12.0% versus 2.9% [with versus without HF], P<0.0001) and at 6 months after discharge (8.5% versus 2.8%, P<0.0001). Of note, HF increased mortality rates in patients with unstable angina (defined as ACS with normal biochemical markers of necrosis; mortality rates: 6.7% with versus 1.6% without HF at admission, P<0.0001). By logistic regression analysis, admission HF was an independent predictor of hospital death (odds ratio, 2.2; P<0.0001). Admission HF was associated with longer hospital stay and higher readmission rates. Patients with HF had lower rates of catheterization and percutaneous cardiac intervention, and fewer received beta-blockers and statins. Hospital development of HF (versus HF on presentation) was associated with an even higher in-hospital mortality rate (17.8% versus 12.0%, P<0.0001). In patients with HF, in-hospital revascularization was associated with lower 6-month death rates (14.0% versus 23.7%, P<0.0001; adjusted hazard ratio, 0.5; 95% CI, 0.37 to 0.68, P<0.0001). CONCLUSIONS: In this observational registry, heart failure was associated with reduced hospital and 6-month survival across all ACS subsets, including patients with normal markers of necrosis. More aggressive treatment of these patients may be warranted to improve prognosis. PMID- 14744971 TI - Inhibition of restenosis with a paclitaxel-eluting, polymer-free coronary stent: the European evaLUation of pacliTaxel Eluting Stent (ELUTES) trial. AB - BACKGROUND: The use of a stent to deliver a drug may reduce in-stent restenosis. Paclitaxel interrupts the smooth muscle cell cycle by stabilizing microtubules, thereby arresting mitosis. METHODS AND RESULTS: On the basis of prior animal studies, the European evaLUation of the pacliTaxel Eluting Stent (ELUTES) pilot clinical trial (n=190) investigated the safety and efficacy of V-Flex Plus coronary stents (Cook Inc) coated with escalating doses of paclitaxel (0.2, 0.7, 1.4, and 2.7 microg/mm2 stent surface area) applied directly to the abluminal surface of the stent in de novo lesions compared with bare stent alone. The primary efficacy end point was angiographic percent diameter stenosis at 6 months. At angiographic follow-up, percent diameter stenosis was 33.9+/-26.7% in controls (n=34) and 14.2+/-16.6% in the 2.7-microg/mm2 group (n=31; P=0.006). Late loss decreased from 0.73+/-0.73 to 0.11+/-0.50 mm (P=0.002). Binary restenosis (> or =50% at follow-up) decreased from 20.6% to 3.2% (P=0.056), with no significant benefit from intermediate paclitaxel doses. Freedom from major adverse cardiac events in the highest (effective) dose group was 92%, 89%, and 86% at 1, 6, and 12 months, respectively (P=NS versus control). No late stent thromboses were seen in any treated group despite clopidogrel treatment for 3 months only. CONCLUSIONS: Paclitaxel applied directly to the abluminal surface of a bare metal coronary stent, at a dose density of 2.7 microg/mm2, reduced angiographic indicators of in-stent restenosis without short- or medium-term side effects. PMID- 14744972 TI - High plasma heparin cofactor II activity is associated with reduced incidence of in-stent restenosis after percutaneous coronary intervention. AB - BACKGROUND: Thrombin plays an important role in the development of atherosclerosis and restenosis after percutaneous coronary intervention. Because heparin cofactor II (HCII) inhibits thrombin action in the presence of dermatan sulfate, which is abundantly present in arterial wall, HCII may affect vascular remodeling by modulating thrombin action. We hypothesized that patients with high plasma HCII activity may show a reduced incidence of in-stent restenosis (ISR). METHODS AND RESULTS: Sequential coronary arteries (n=166) with NIR stent (Boston Scientific Corp) implantation in 134 patients were evaluated before, immediately after, and at 6 months after percutaneous coronary intervention. Patients were divided into the following groups: high HCII (> or =110%, 45 lesions in 36 patients), normal HCII (> or =80% and <110%, 81 lesions in 66 patients), and low HCII (<80%, 40 lesions in 32 patients). Percent diameter stenosis at follow-up in the high-HCII group (18.7%) was significantly lower (P=0.046) than that in the normal-HCII group (30.3%) or the low-HCII group (29.0%). The ISR rate in the high HCII group (6.7%) was significantly lower than that in the low-HCII group (30.0%) (P=0.0039). Furthermore, multivariate analysis demonstrated that high plasma HCII activity is an independent factor in reducing the incidence of angiographic restenosis (odds ratio, 0.953/1% increase of HCII; 95% CI, 0.911 to 0.998). CONCLUSIONS: The results demonstrate that HCII may have a hitherto unrecognized effect in inhibiting ISR. The effect of HCII may be mediated by inactivating thrombin in injured arteries, thereby inhibiting vascular smooth muscle cell migration and proliferation. PMID- 14744973 TI - Vascular oxidant stress enhances progression and angiogenesis of experimental atheroma. AB - BACKGROUND: Although multiple pathological processes have been associated with oxidative stress, the causative relation between oxidative stress and arterial lesion progression remains unclear. METHODS AND RESULTS: To test the effect of creating arterial wall oxidative stress, we compared progression of mouse carotid lesions induced by flow cessation in the wild-type (WT) versus transgenic mice (Tg(p22vsmc)), in which overexpression of p22phox, a critical component of NAD(P)H oxidase was targeted to smooth muscle cell (SMC). Compared with WT mice, arterial lesions grew significantly larger in Tg(p22vsmc) (P<0.001) and demonstrated elevated hydrogen peroxide (H2O2) and vascular endothelial growth factor (VEGF) levels at all time points examined (P<0.001, n=4 animals per time point), probably related to increased expression of hypoxia inducible factor (HIF)-1alpha via SMC oxidative stress in the Tg(p22vsmc) arteries, both basally (203+/-12% versus WT, P<0.001, n=3) and after lesion formation. Interestingly, Tg(p22vsmc) lesions were complicated by extensive neointimal angiogenesis. In vitro experiments confirmed SMCs isolated from Tg(p22vsmc) to be the source for increased H2O2, VEGF, and HIF-1alpha and their capacity to induce angiogenic cord like structures when cocultured with endothelial cells. The antioxidant ebselen inhibited SMC activities in vitro and intralesion angiogenesis and lesion progression in vivo. CONCLUSIONS: We have demonstrated a novel pathway by which oxidative stress can trigger in vivo an angiogenic switch associated with experimental plaque progression and angiogenesis. This pathway may be related to human atheroma progression and destabilization through intraplaque hemorrhage. PMID- 14744974 TI - Overexpression of glutathione peroxidase prevents left ventricular remodeling and failure after myocardial infarction in mice. AB - BACKGROUND: Oxidative stress plays an important role in the pathophysiology of heart failure. We determined whether the overexpression of glutathione peroxidase (GSHPx) could attenuate left ventricular (LV) remodeling and failure after myocardial infarction (MI). METHODS AND RESULTS: We created MI in 12- to 16-week old, male GSHPx transgenic mice (TG+MI) and nontransgenic wild-type littermates (WT+MI) by ligating the left coronary artery. GSHPx activity was increased in the hearts of TG mice, with no significant changes in other antioxidant enzymes. LV concentrations of thiobarbituric acid-reactive substances measured in TG+MI at 4 weeks were significantly lower than those in WT+MI. The survival rate during 4 weeks of MI was significantly higher in TG+MI than in WT+MI, although the infarct size was comparable. LV cavity dilatation and dysfunction were significantly attenuated in TG+MI. LV end-diastolic pressure was increased in WT+MI and reduced in TG+MI. Improvement of LV function in TG+MI was accompanied by a decrease in myocyte hypertrophy, apoptosis, and interstitial fibrosis in the noninfarcted LV. Myocardial matrix metalloproteinase-9 zymographic and protein levels were increased in WT+MI after 3 days but were attenuated in TG+MI. CONCLUSIONS: Overexpression of GSHPx inhibited LV remodeling and failure after MI. Therapies designed to interfere with oxidative stress might be beneficial to prevent cardiac failure. PMID- 14744975 TI - C-reactive protein accelerates the progression of atherosclerosis in apolipoprotein E-deficient mice. AB - BACKGROUND: Plasma C-reactive protein (CRP) concentration is a strong predictor of atherosclerosis. However, to date, there is no in vivo evidence that CRP is proatherogenic. METHODS AND RESULTS: We studied the effect of human CRP transgene (tg) expression, under basal and turpentine-stimulated conditions, on atherosclerosis in apolipoprotein (apo) E-/- mice. Aortic atherosclerotic lesions in 29-week-old male mice were 48% larger (P<0.02) in turpentine-treated mice and 34% larger (P<0.05) in untreated CRPtg+/0/apoE-/- mice. Turpentine treatment per se did not affect the extent of atherosclerosis in CRP transgenic or nontransgenic apoE-/- mice. Transgenic mice exhibited lower plasma complement C3 but increased deposition of CRP and C3 in the lesions, which suggests that CRP stimulated activation of complement within the lesion. There was more intense and widespread vascular cell adhesion molecule-1 and collagen staining in the lesions of CRPtg+/0/apoE-/- mice than in CRPtg0/0/apoE-/- littermates. Lesions of CRPtg+/0/apoE-/- mice contained increased angiotensin type 1 receptor (AT1-R) transcripts and displayed increased AT1-R immunostaining compared with those of CRPtg0/0/apoE-/- mice. There was no difference in blood pressure in the 2 types of mice, which indicates that the proatherogenic effect of CRP-associated AT1-R overexpression is local and not mediated by its hypertensive properties. CONCLUSIONS: Human CRP transgene expression causes accelerated aortic atherosclerosis in apoE-/- mice. CRP was detected in the lesion, which was associated with increased C3 deposition and increased AT1-R, vascular cell adhesion molecule-1, and collagen expression. These data document a proatherogenic role for CRP in vivo. PMID- 14744976 TI - Localized hypersensitivity and late coronary thrombosis secondary to a sirolimus eluting stent: should we be cautious? AB - BACKGROUND: The US Food and Drug Administration recently issued a warning of subacute thrombosis and hypersensitivity reactions to sirolimus-eluting stents (Cypher). The cause and incidence of these events have not been determined. METHODS AND RESULTS: We present findings of a 58-year-old man who died of late stent thrombosis 18 months after receiving 2 Cypher stents for unstable angina. Although angiographic and intravascular ultrasound results at 8 months demonstrated the absence of neointimal formation, vessel enlargement was present. An autopsy showed aneurysmal dilation of the stented arterial segments with a severe localized hypersensitivity reaction consisting predominantly of T lymphocytes and eosinophils. CONCLUSIONS: The known pharmacokinetic elution profile of Cypher stents and the presence of polymer fragments surrounded by giant cells and eosinophils suggest that a reaction to the polymer may have caused late stent thrombosis. Careful long-term follow-up of patients with vessel enlargement after Cypher stent placement is recommended. PMID- 14744977 TI - Cassette-like variation of restriction enzyme genes in Escherichia coli C and relatives. AB - A surprising result of comparative bacterial genomics has been the large amount of DNA found to be present in one strain but not in another of the same species. We examine in detail one location where gene content varies extensively, the restriction cluster in Escherichia coli. This region is designated the Immigration Control Region (ICR) for the density and variability of restriction functions found there. To better define the boundaries of this variable locus, we determined the sequence of the region from a restrictionless strain, E.coli C. Here we compare the 13.7 kb E.coli C sequence spanning the site of the ICR with corresponding sequences from five E.coli strains and Salmonella typhimurium LT2. To discuss this variation, we adopt the term 'framework' to refer to genes that are stable components of genomes within related lineages, while 'migratory' genes are transient inhabitants of the genome. Strikingly, seven different migratory DNA segments, encoding different sets of genes and gene fragments, alternatively occupy a single well-defined location in the seven strains examined. The flanking framework genes, yjiS and yjiA, display approximately normal patterns of conservation. The patterns observed are consistent with the action of a site specific recombinase. Since no nearby gene codes for a likely recombinase of known families, such a recombinase must be of a new family or unlinked. PMID- 14744978 TI - Design and synthesis of a photocleavable biotinylated nucleotide for DNA analysis by mass spectrometry. AB - We report here the design, synthesis and evaluation of a novel photocleavable (PC) biotinylated nucleotide analog, dUTP-PC-Biotin, for DNA polymerase extension reaction to isolate DNA products for mass spectrometry (MS) analysis. This nucleotide analog has a biotin moiety attached to the 5-position of 2' deoxyribouridine 5'-triphosphate via a photocleavable 2-nitrobenzyl linker. We have demonstrated that dUTP-PC-Biotin can be faithfully incorporated by the DNA polymerase Thermo Sequenase into the growing DNA strand in a DNA polymerase extension reaction and that its incorporation does not hinder the addition of the subsequent nucleotide. Therefore, the DNA extension fragments generated by using the dUTP-PC-Biotin can be efficiently isolated by a streptavidin-coated surface and recovered by near-UV light irradiation at room temperature in mild condition for further analysis without using any chemicals or heat. Single and multiple primer extension reactions were performed using the dUTP-PC-Biotin to generate DNA products for MALDI-TOF MS analysis. Such nucleotide analogs that carry a biotin and a photocleavable linker will allow the isolation and purification of DNA products under mild conditions for MS-based genetic analysis by DNA sequencing or multiplex single nucleotide polymorphism (SNP) detection. Furthermore, these nucleotide analogs should also be useful in isolating DNA protein complexes under non-denaturing conditions. PMID- 14744979 TI - Surprising features of plastid ndhD transcripts: addition of non-encoded nucleotides and polysome association of mRNAs with an unedited start codon. AB - RNA editing in higher plant plastids is a post- transcriptional RNA maturation process changing single cytidine nucleotides into uridine. In the ndhD transcript of tobacco and several other plant species, editing of an ACG codon to a standard AUG initiator codon is believed to be a prerequisite for translation. In order to test this assumption experimentally, we have analyzed the editing status of ndhD mRNA species in the process of translation. We show that unedited ndhD transcripts are also associated with polysomes in vivo, suggesting that they are translated. This surprising finding challenges the view that ACG to AUG editing is strictly required to make the ndhD message translatable and raises the possibility that ACG can be utilized as an initiator codon in chloroplasts. In addition, we have mapped the termini of the ndhD transcript and discovered a novel form of RNA processing. Unexpectedly, we find that highly specific sequences are added to the 3' end of the ndhD mRNA at high frequency. We propose a model in which these sequences are added by the successive action of a CCA adding enzyme (tRNA nucleotidyltransferase) and an RNA-dependent RNA polymerase (RdRp) activity. The presence of an RdRp activity may have general implications also for other steps in plastid gene expression. PMID- 14744980 TI - Protein structure prediction using sparse dipolar coupling data. AB - Residual dipolar coupling (RDC) represents one of the most exciting emerging NMR techniques for protein structure studies. However, solving a protein structure using RDC data alone is still a highly challenging problem. We report here a computer program, RDC-PROSPECT, for protein structure prediction based on a structural homolog or analog of the target protein in the Protein Data Bank (PDB), which best aligns with the (15)N-(1)H RDC data of the protein recorded in a single ordering medium. Since RDC-PROSPECT uses only RDC data and predicted secondary structure information, its performance is virtually independent of sequence similarity between a target protein and its structural homolog/analog, making it applicable to protein targets beyond the scope of current protein threading techniques. We have tested RDC-PROSPECT on all (15)N-(1)H RDC data (representing 43 proteins) deposited in the BioMagResBank (BMRB) database. The program correctly identified structural folds for 83.7% of the target proteins, and achieved an average alignment accuracy of 98.1% residues within a four residue shift. PMID- 14744981 TI - Automated correction of genome sequence errors. AB - By using information from an assembly of a genome, a new program called AutoEditor significantly improves base calling accuracy over that achieved by previous algorithms. This in turn improves the overall accuracy of genome sequences and facilitates the use of these sequences for polymorphism discovery. We describe the algorithm and its application in a large set of recent genome sequencing projects. The number of erroneous base calls in these projects was reduced by 80%. In an analysis of over one million corrections, we found that AutoEditor made just one error per 8828 corrections. By substantially increasing the accuracy of base calling, AutoEditor can dramatically accelerate the process of finishing genomes, which involves closing all gaps and ensuring minimum quality standards for the final sequence. It also greatly improves our ability to discover single nucleotide polymorphisms (SNPs) between closely related strains and isolates of the same species. PMID- 14744983 TI - The 57th Annual Meeting and Symposium of the Society of General Physiologists: the biology of chloride. PMID- 14744982 TI - Superior 5' homogeneity of RNA from ATP-initiated transcription under the T7 phi 2.5 promoter. AB - Transcription from the commonly used GTP- initiating T7 class III promoter phi6.5 frequently produces heterogeneous RNA at both 3' and 5' ends. We demonstrate here that RNA transcripts from the T7 class II promoter phi2.5 have superior 5' homogeneity over those from the phi6.5 promoter, with comparable total RNA yields. The overall homogeneity of RNA transcripts is improved to different degrees depending on RNA sequences, although transcription under phi2.5 does not affect the 3' heterogeneity of RNA. In combination with 3' RNA trimming by DNAzymes or ribozymes, this ATP- initiated transcription system based on the T7 phi2.5 promoter can provide excellent quality of RNA for applications requiring a high degree of RNA size homogeneity. PMID- 14744984 TI - Paul F. Cranefield Award to Daniel H. Cox. PMID- 14744985 TI - What the frog's nose tells the frog's brain. PMID- 14744986 TI - Classes and narrowing selectivity of olfactory receptor neurons of Xenopus laevis tadpoles. AB - In olfactory receptor neurons (ORNs) of aquatic animals amino acids have been shown to be potent stimuli. Here we report on calcium imaging experiments in slices of the olfactory mucosa of Xenopus laevis tadpoles. We were able to determine the response profiles of 283 ORNs to 19 amino acids, where one profile comprises the responses of one ORN to 19 amino acids. 204 out of the 283 response profiles differed from each other. 36 response spectra occurred more than once, i.e., there were 36 classes of ORNs identically responding to the 19 amino acids. The number of ORNs that formed a class ranged from 2 to 13. Shape and duration of amino acid-elicited [Ca2+]i transients showed a high degree of similarity upon repeated stimulation with the same amino acid. Different amino acids, however, in some cases led to clearly distinguishable calcium responses in individual ORNs. Furthermore, ORNs clearly appeared to gain selectivity over time, i.e., ORNs of later developmental stages responded to less amino acids than ORNs of earlier stages. We discuss the narrowing of ORN selectivity over stages in the context of expression of olfactory receptors. PMID- 14744987 TI - Membrane tension accelerates rate-limiting voltage-dependent activation and slow inactivation steps in a Shaker channel. AB - A classical voltage-sensitive channel is tension sensitive--the kinetics of Shaker and S3-S4 linker deletion mutants change with membrane stretch (Tabarean, I.V., and C.E. Morris. 2002. Biophys. J. 82:2982-2994.). Does stretch distort the channel protein, producing novel channel states, or, more interestingly, are existing transitions inherently tension sensitive? We examined stretch and voltage dependence of mutant 5aa, whose ultra-simple activation (Gonzalez, C., E. Rosenman, F. Bezanilla, O. Alvarez, and R. Latorre. 2000. J. Gen. Physiol. 115:193-208.) and temporally matched activation and slow inactivation were ideal for these studies. We focused on macroscopic patch current parameters related to elementary channel transitions: maximum slope and delay of current rise, and time constant of current decline. Stretch altered the magnitude of these parameters, but not, or minimally, their voltage dependence. Maximum slope and delay versus voltage with and without stretch as well as current rising phases were well described by expressions derived for an irreversible four-step activation model, indicating there is no separate stretch-activated opening pathway. This model, with slow inactivation added, explains most of our data. From this we infer that the voltage-dependent activation path is inherently stretch sensitive. Simulated currents for schemes with additional activation steps were compared against datasets; this showed that generally, additional complexity was not called for. Because the voltage sensitivities of activation and inactivation differ, it was not possible to substitute depolarization for stretch so as to produce the same overall PO time course. What we found, however, was that at a given voltage, stretch-accelerated current rise and decline almost identically--normalized current traces with and without stretch could be matched by a rescaling of time. Rate-limitation of the current falling phase by activation was ruled out. We hypothesize, therefore, that stretch-induced bilayer decompression facilitates an in-plane expansion of the protein in both activation and inactivation. Dynamic structural models of this class of channels will need to take into account the inherent mechanosensitivity of voltage-dependent gating. PMID- 14744988 TI - The Na+ channel inactivation gate is a molecular complex: a novel role of the COOH-terminal domain. AB - Electrical activity in nerve, skeletal muscle, and heart requires finely tuned activity of voltage-gated Na+ channels that open and then enter a nonconducting inactivated state upon depolarization. Inactivation occurs when the gate, the cytoplasmic loop linking domains III and IV of the alpha subunit, occludes the open pore. Subtle destabilization of inactivation by mutation is causally associated with diverse human disease. Here we show for the first time that the inactivation gate is a molecular complex consisting of the III-IV loop and the COOH terminus (C-T), which is necessary to stabilize the closed gate and minimize channel reopening. When this interaction is disrupted by mutation, inactivation is destabilized allowing a small, but important, fraction of channels to reopen, conduct inward current, and delay cellular repolarization. Thus, our results demonstrate for the first time that physiologically crucial stabilization of inactivation of the Na+ channel requires complex interactions of intracellular structures and indicate a novel structural role of the C-T domain in this process. PMID- 14744989 TI - A store-operated calcium channel in Drosophila S2 cells. AB - Using whole-cell recording in Drosophila S2 cells, we characterized a Ca(2+) selective current that is activated by depletion of intracellular Ca2+ stores. Passive store depletion with a Ca(2+)-free pipette solution containing 12 mM BAPTA activated an inwardly rectifying Ca2+ current with a reversal potential >60 mV. Inward currents developed with a delay and reached a maximum of 20-50 pA at 110 mV. This current doubled in amplitude upon increasing external Ca2+ from 2 to 20 mM and was not affected by substitution of choline for Na+. A pipette solution containing approximately 300 nM free Ca2+ and 10 mM EGTA prevented spontaneous activation, but Ca2+ current activated promptly upon application of ionomycin or thapsigargin, or during dialysis with IP3. Isotonic substitution of 20 mM Ca2+ by test divalent cations revealed a selectivity sequence of Ba2+ > Sr2+ > Ca2+ >> Mg2+. Ba2+ and Sr2+ currents inactivated within seconds of exposure to zero-Ca2+ solution at a holding potential of 10 mV. Inactivation of Ba2+ and Sr2+ currents showed recovery during strong hyperpolarizing pulses. Noise analysis provided an estimate of unitary conductance values in 20 mM Ca2+ and Ba2+ of 36 and 420 fS, respectively. Upon removal of all external divalent ions, a transient monovalent current exhibited strong selectivity for Na+ over Cs+. The Ca2+ current was completely and reversibly blocked by Gd3+, with an IC50 value of approximately 50 nM, and was also blocked by 20 microM SKF 96365 and by 20 microM 2-APB. At concentrations between 5 and 14 microM, application of 2-APB increased the magnitude of Ca2+ currents. We conclude that S2 cells express store-operated Ca2+ channels with many of the same biophysical characteristics as CRAC channels in mammalian cells. PMID- 14744992 TI - Asymmetric division and lineage commitment at the level of hematopoietic stem cells: inference from differentiation in daughter cell and granddaughter cell pairs. AB - How hematopoietic stem cells (HSCs) commit to a particular lineage is unclear. A high degree of HSC purification enabled us to address this issue at the clonal level. Single-cell transplantation studies revealed that 40% of the CD34-/low, c Kit+, Sca-1+, and lineage marker- (CD34-KSL) cells in adult mouse bone marrow were able, as individual cells, to reconstitute myeloid and B- and T-lymphoid lineages over the long-term. Single-cell culture showed that >40% of CD34-KSL cells could form neutrophil (n)/macrophage (m)/erythroblast (E)/megakaryocyte (M) (nmEM) colonies. Assuming that a substantial portion of long-term repopulating cells can be detected as nmEM cells within this population, we compared differentiation potentials between individual pairs of daughter and granddaughter cells derived in vitro from single nmEM cells. One of the two daughter or granddaughter cells remained an nmEM cell. The other showed a variety of combinations of differentiation potential. In particular, an nmEM cell directly gave rise, after one cell division, to progenitor cells committed to nm, EM, or M lineages. The probability of asymmetric division of nmEM cells depended on the cytokines used. These data strongly suggest that lineage commitment takes place asymmetrically at the level of HSCs under the influence of external factors. PMID- 14744991 TI - Insulin induces the release of vasodilator compounds from platelets by a nitric oxide-G kinase-VAMP-3-dependent pathway. AB - Insulin-induced vasodilatation is sensitive to nitric oxide (NO) synthase (NOS) inhibitors. However, insulin is unable to relax isolated arteries or to activate endothelial NOS in endothelial cells. Since insulin can enhance platelet endothelial NOS activity, we determined whether insulin-induced vasodilatation can be attributed to a NO-dependent, platelet-mediated process. Insulin failed to relax endothelium-intact rings of porcine coronary artery. The supernatant from insulin-stimulated human platelets induced complete relaxation, which was prevented by preincubation of platelets with a NOS inhibitor, the soluble guanylyl cyclase inhibitor, NS 2028, or the G kinase inhibitor, KT 5823, and was abolished by an adenosine A2A receptor antagonist. Insulin induced the release of adenosine trisphosphate (ATP), adenosine, and serotonin from platelet-dense granules in a NO-dependent manner. This response was not detected using insulin stimulated platelets from endothelial NOS-/- mice, although a NO donor elicited ATP release. Insulin-induced ATP release from human platelets correlated with the association of syntaxin 2 with the vesicle-associated membrane protein 3 but was not associated with the activation of alphaIIbbeta3 integrin. Thus, insulin elicits the release of vasoactive concentrations of ATP and adenosine from human platelets via a NO-G kinase-dependent signaling cascade. The mechanism of dense granule secretion involves the G kinase-dependent association of syntaxin 2 with vesicle-associated membrane protein 3. PMID- 14744993 TI - Chemokine receptor CCR9 contributes to the localization of plasma cells to the small intestine. AB - Humoral immunity in the gut-associated lymphoid tissue is characterized by the production of immunoglobulin A (IgA) by antibody-secreting plasma cells (PCs) in the lamina propria. The chemokine CCL25 is expressed by intestinal epithelial cells and is capable of inducing chemotaxis of IgA+ PCs in vitro. Using a newly generated monoclonal antibody against murine CCR9, we show that IgA+ PCs express high levels of CCR9 in the mesenteric lymph node (MLN) and Peyer's patches (PPs), but down-regulate CCR9 once they are located in the small intestine. In CCR9 deficient mice, IgA+ PCs are substantially reduced in number in the lamina propria of the small intestine. In adoptive transfer experiments, CCR9-deficient IgA+ PCs show reduced migration into the small intestine compared with wild-type controls. Furthermore, CCR9 mutants fail to mount a regular IgA response to an orally administered antigen, although the architecture and cell type composition of PPs and MLN are unaffected and are functional for the generation of IgA PCs. These findings provide profound in vivo evidence that CCL25/CCR9 guides PCs into the small intestine. PMID- 14744994 TI - CD1d-dependent activation of NKT cells aggravates atherosclerosis. AB - Adaptive and innate immunity have been implicated in the pathogenesis of atherosclerosis. Given their abundance in the lesion, lipids might be targets of the atherosclerosis-associated immune response. Natural killer T (NKT) cells can recognize lipid antigens presented by CD1 molecules. We have explored the role of CD1d-restricted NKT cells in atherosclerosis by using apolipoprotein E-deficient (apoE-/-) mice, a hypercholesterolemic mouse model that develops atherosclerosis. ApoE-/- mice crossed with CD1d-/- (CD1d-/-apoE-/-) mice exhibited a 25% decrease in lesion size compared with apoE-/- mice. Administration of alpha galactosylceramide, a synthetic glycolipid that activates NKT cells via CD1d, induced a 50% increase in lesion size in apoE-/- mice, whereas it did not affect lesion size in apoE-/-CD1d-/- mice. Treatment was accompanied by an early burst of cytokines (IFNgamma, MCP-1, TNFalpha, IL-2, IL-4, IL-5, and IL-6) followed by sustained increases in IFNgamma and IL-4 transcripts in the spleen and aorta. Early activation of both T and B cells was followed by recruitment of T and NKT cells to the aorta and activation of inflammatory genes. These results show that activation of CD1d-restricted NKT cells exacerbates atherosclerosis. PMID- 14744995 TI - Massive thymic deletion results in systemic autoimmunity through elimination of CD4+ CD25+ T regulatory cells. AB - Incomplete deletion of KRN T cells that recognize the ubiquitously expressed self antigen glucose-6-phosphate-isomerase (GPI) initiates an anti-GPI autoimmune cascade in K/BxN mice resulting in a humorally mediated arthritis. Transgenic (Tg) expression of a KRN T cell receptor (TCR) agonist under the major histocompatibility complex class II promoter resulted in thymic deletion with loss of anti-GPI T and B cell responses and attenuated arthritis course. However, double Tg mice succumbed to systemic autoimmunity with multiorgan inflammation and autoantibody production. Extensive thymic deletion resulted in lymphopenia and elimination of CD4+ CD25+ regulatory T cells (Tregs), but spared some CD4+ T cells expressing endogenous TCR, which oligoclonally expanded in the periphery. Disease was transferred by these T cells and prevented by cotransfer of CD4+ CD25+ Tregs. Moreover, we extended our findings to another TCR system (anti-hen egg lysozyme [HEL] TCR/HEL mice) where similarly extensive thymic deletion also resulted in disease. Thus, our studies demonstrated that central tolerance can paradoxically result in systemic autoimmunity through differential susceptibility of Tregs and autoreactive T cells to thymic deletion. Therefore, too little or too much negative selection to a self-antigen can result in systemic autoimmunity and disease. PMID- 14744996 TI - The metallo-beta-lactamase/beta-CASP domain of Artemis constitutes the catalytic core for V(D)J recombination. AB - The V(D)J recombination/DNA repair factor Artemis belongs to the metallo-beta lactamase (beta-Lact) superfamily of enzymes. Three regions can be defined within the Artemis protein sequence: (a) the beta-Lact homology domain, to which is appended (b) the beta-CASP region, specific of members of the beta-Lact superfamily acting on nucleic acids, and (c) the COOH-terminal domain. Using in vitro mutagenesis, here we show that the association of the beta-Lact and the beta-CASP regions suffices for in vivo V(D)J recombination of chromosome integrated substrates. Single amino acid mutants point to critical catalytic residues for V(D)J recombination activity. The results presented here define the beta-Lact/beta-CASP domain of Artemis as the minimal core catalytic domain needed for V(D)J recombination and suggest that Artemis uses one or two Zn(II) ions to exert its catalytic activity, like bacterial class B beta-Lact enzymes hydrolyzing beta-lactam compounds. PMID- 14744997 TI - Extracellular HMGB1, a signal of tissue damage, induces mesoangioblast migration and proliferation. AB - High mobility group box 1 (HMGB1) is an abundant chromatin protein that acts as a cytokine when released in the extracellular milieu by necrotic and inflammatory cells. Here, we show that extracellular HMGB1 and its receptor for advanced glycation end products (RAGE) induce both migration and proliferation of vessel associated stem cells (mesoangioblasts), and thus may play a role in muscle tissue regeneration. In vitro, HMGB1 induces migration and proliferation of both adult and embryonic mesoangioblasts, and disrupts the barrier function of endothelial monolayers. In living mice, mesoangioblasts injected into the femoral artery migrate close to HMGB1-loaded heparin-Sepharose beads implanted in healthy muscle, but are unresponsive to control beads. Interestingly, alpha-sarcoglycan null dystrophic muscle contains elevated levels of HMGB1; however, mesoangioblasts migrate into dystrophic muscle even if their RAGE receptor is disabled. This implies that the HMGB1-RAGE interaction is sufficient, but not necessary, for mesoangioblast homing; a different pathway might coexist. Although the role of endogenous HMGB1 in the reconstruction of dystrophic muscle remains to be clarified, injected HMGB1 may be used to promote tissue regeneration. PMID- 14744999 TI - Signaling switches and bistability arising from multisite phosphorylation in protein kinase cascades. AB - Mitogen-activated protein kinase (MAPK) cascades can operate as bistable switches residing in either of two different stable states. MAPK cascades are often embedded in positive feedback loops, which are considered to be a prerequisite for bistable behavior. Here we demonstrate that in the absence of any imposed feedback regulation, bistability and hysteresis can arise solely from a distributive kinetic mechanism of the two-site MAPK phosphorylation and dephosphorylation. Importantly, the reported kinetic properties of the kinase (MEK) and phosphatase (MKP3) of extracellular signal-regulated kinase (ERK) fulfill the essential requirements for generating a bistable switch at a single MAPK cascade level. Likewise, a cycle where multisite phosphorylations are performed by different kinases, but dephosphorylation reactions are catalyzed by the same phosphatase, can also exhibit bistability and hysteresis. Hence, bistability induced by multisite covalent modification may be a widespread mechanism of the control of protein activity. PMID- 14744998 TI - WASp is required for the correct temporal morphogenesis of rhabdomere microvilli. AB - Microvilli are actin-based fingerlike membrane projections that form the basis of the brush border of enterocytes and the Drosophila melanogaster photoreceptor rhabdomere. Although many microvillar cytoskeletal components have been identified, the molecular basis of microvillus formation is largely undefined. Here, we report that the Wiskott-Aldrich syndrome protein (WASp) is necessary for rhabdomere microvillus morphogenesis. We show that WASp accumulates on the photoreceptor apical surface before microvillus formation, and at the time of microvillus initiation WASp colocalizes with amphiphysin and moesin. The loss of WASp delays the enrichment of F-actin on the apical photoreceptor surface, delays the appearance of the primordial microvillar projections, and subsequently leads to malformed rhabdomeres. PMID- 14745000 TI - Mating type-dependent constraints on the mobility of the left arm of yeast chromosome III. AB - Mating-type gene (MAT) switching in budding yeast exhibits donor preference. MATa preferentially recombines with HML near the left telomere of chromosome III, whereas MATalpha prefers HMR near the right telomere. Donor preference is controlled by the recombination enhancer (RE) located proximal to HML. To test if HML is constrained in pairing with MATalpha, we examined live-cell mobility of LacI-GFP-bound lactose operator (lacO) arrays inserted at different chromosomal sites. Without induction of recombination, lacO sequences adjacent to HML are strongly constrained in both MATalpha and RE-deleted MATa strains, compared with MATa. In contrast, chromosome movement at HMR or near a telomere of chromosome V is mating-type independent. HML is more constrained in MATa Deltare and less constrained in MATa RE+ compared with other sites. Although HML and MATa are not prealigned before inducing recombination, the three-dimensional configuration of MAT, HML, and HMR is mating-type dependent. These data suggest there is constitutive tethering of HML, which is relieved in MATa cells through the action of RE. PMID- 14745001 TI - Noninvasive two-photon imaging reveals retinyl ester storage structures in the eye. AB - Visual sensation in vertebrates is triggered when light strikes retinal photoreceptor cells causing photoisomerization of the rhodopsin chromophore 11 cis-retinal to all-trans-retinal. The regeneration of preillumination conditions of the photoreceptor cells requires formation of 11-cis-retinal in the adjacent retinal pigment epithelium (RPE). Using the intrinsic fluorescence of all-trans retinyl esters, noninvasive two-photon microscopy revealed previously uncharacterized structures (6.9 +/- 1.1 microm in length and 0.8 +/- 0.2 microm in diameter) distinct from other cellular organelles, termed the retinyl ester storage particles (RESTs), or retinosomes. These structures form autonomous all trans-retinyl ester-rich intracellular compartments distinct from other organelles and colocalize with adipose differentiation-related protein. As demonstrated by in vivo experiments using wild-type mice, the RESTs participate in 11-cis-retinal formation. RESTs accumulate in Rpe65-/- mice incapable of carrying out the enzymatic isomerization, and correspondingly, are absent in the eyes of Lrat-/- mice deficient in retinyl ester synthesis. These results indicate that RESTs located close to the RPE plasma membrane are essential components in 11-cis-retinal production. PMID- 14745002 TI - Defective angiogenesis and fatal embryonic hemorrhage in mice lacking core 1 derived O-glycans. AB - The core 1 beta1-3-galactosyltransferase (T-synthase) transfers Gal from UDP-Gal to GalNAcalpha1-Ser/Thr (Tn antigen) to form the core 1 O-glycan Galbeta1 3GalNAcalpha1-Ser/Thr (T antigen). The T antigen is a precursor for extended and branched O-glycans of largely unknown function. We found that wild-type mice expressed the NeuAcalpha2-3Galbeta1-3GalNAcalpha1-Ser/Thr primarily in endothelial, hematopoietic, and epithelial cells during development. Gene targeted mice lacking T-synthase instead expressed the nonsialylated Tn antigen in these cells and developed brain hemorrhage that was uniformly fatal by embryonic day 14. T-synthase-deficient brains formed a chaotic microvascular network with distorted capillary lumens and defective association of endothelial cells with pericytes and extracellular matrix. These data reveal an unexpected requirement for core 1-derived O-glycans during angiogenesis. PMID- 14745003 TI - Coat protein regulates formation of replication complexes during tobacco mosaic virus infection. AB - The genome of tobacco mosaic virus (TMV) encodes replicase protein(s), movement protein (MP), and capsid protein (CP). On infection, one or more viral proteins direct the assembly of virus replication complexes (VRCs), in association with host-derived membranes. The impact of CP-mediated resistance on the structures of the replication complexes was examined in nontransgenic and transgenic BY-2 cell lines that produce wild-type CP, mutant CP(T42W), and Ds-Red, which was targeted to endoplasmic reticulum by using immunofluorescence and 3D microscopy. We developed a model of VRCs that shows a clear association of MP with and surrounding the endoplasmic reticulum. Replicase is located within the MP bodies, as well as isolated sites throughout the cell. CP surrounds the VRCs. CP enhances the production of MP and increases the size of the VRC; however, the mutant CP(T42W) reduces the amount of MP and interferes with the formation of VRCs. We propose a regulatory role of the CP in the establishment of the VRC. We suggest that the lack of formation of VRCs restricts the efficiency of virus replication and the formation of virus movement complexes, resulting in restriction of cell cell spread of infection. This results in higher levels of plant CP-mediated protection provided by CP(T42W). PMID- 14745004 TI - ELG1, a regulator of genome stability, has a role in telomere length regulation and in silencing. AB - Telomeres, the natural ends of eukaryotic chromosomes, prevent the loss of chromosomal sequences and preclude their recognition as broken DNA. Telomere length is kept under strict boundaries by the action of various proteins, some with negative and others with positive effects on telomere length. Recently, data have been accumulating to support a role for DNA replication in the control of telomere length, although through a currently poorly understood mechanism. Elg1p, a replication factor C (RFC)-like protein of yeast, contributes to genome stability through a putative replication-associated function. Here, we show that Elg1p participates in negative control of telomere length and in telomeric silencing through a replication-mediated pathway. We show that the telomeric function of Elg1 is independent of recombination and completely dependent on an active telomerase. Additionally, this function depends on yKu and DNA polymerase. We discuss alternative models to explain how Elg1p affects telomere length. PMID- 14745005 TI - A pituitary gene encodes a protein that produces differentiation of breast and prostate cancer cells. AB - A cDNA clone of 1.1 kb encoding a 108-aa polypeptide was isolated from a human pituitary cDNA library by expression cloning. This protein was named tumor differentiation factor (TDF). The recombinant TDF protein and a 20-aa peptide, P1, selected from the ORF of the gene, induced morphological and biochemical changes consistent with differentiation of human breast and prostate cancer cells. Fibroblast, kidney, hepatoma, and leukemic lymphocytic cell lines were unaffected. Breast and prostate cancer cells aggregated in spheroid-like structures within 24 h of exposure to TDF. This effect was abrogated by a specific affinity-purified rabbit polyclonal anti-P1 Ab. E-cadherin expression was increased in a dose-dependent manner by TDF. Treatment of MCF7 cells with TDF led to production of a lactalbumin-related protein. Peptide P1 significantly decreased the growth of androgen-independent DU145 prostate cancer in severe combined immunodeficient mice. The presence of TDF protein in human sera was detected by the anti-P1 Ab, suggesting a role of TDF in endocrine metabolism. The fact that all activities of TDF can be mimicked by a peptide derived from the encoding TDF sequence opens the possibility of therapeutic applications. PMID- 14745006 TI - Autoimmune glomerulonephritis with spontaneous formation of splenic germinal centers in mice lacking the estrogen receptor alpha gene. AB - In mice, ovariectomy accelerates the progression of the end-stage renal disease glomerulosclerosis. In women, the incidence of this disease increases after menopause, and estrogen alters its progression. Polymorphisms in the human estrogen receptor alpha (ERalpha) gene have been suggested to constitute a genetic predisposition for lupus nephritis. Here we show that by 1 year of age, mice lacking ERalpha (ERalpha(-/-)) but not those lacking ERbeta (ERbeta(-/-)) exhibit immune complex-type glomerulonephritis, proteinuria, and destruction of tubular cells with severe infiltration of B lymphocytes in the kidney and the presence of anti-DNA antibodies in serum. No gender difference occurred in the incidence or severity of these symptoms. However, in female but not in male ERalpha(-/-) mice there were elevated serum levels of IgG3. Other prominent features of these mice were (i) spontaneous formation of germinal centers in the spleen in the absence of antigen challenge and (ii) infiltration of plasma cells in the kidney and plasmacytosis in the spleen. Immunohistochemistry indicated a selective expression of ERalpha protein in the germinal centers but not in the follicular mantle zone of murine spleens and human tonsils. Our results indicate that ERalpha has indispensable functions in the kidney and in germinal centers, and that defective ERalpha signaling results in glomerulonephritis. PMID- 14745007 TI - Cerebellar granule cell precursors can differentiate into astroglial cells. AB - During CNS development, multipotent neural stem cells give rise first to various kinds of specified precursor cells, which proliferate extensively before terminally differentiating into either neurons or glial cells. It is still not clear, however, whether the specified precursor cells are irreversibly determined to differentiate into their particular cell types. In this study, we show that isolated mouse cerebellar granule cell precursors from the outermost, proliferative zone of the external germinal layer can differentiate into astroglial cells when exposed to sonic hedgehog (Shh) and bone morphogenetic proteins. These induced cells initially expressed both glial fibrillary acidic protein and neuronal markers, but they then lost their neuronal markers and acquired S100-beta, a marker of differentiated astroglial cells. These results indicate that at least some granule cell precursors are not irreversibly committed to neuronal development but can be induced to differentiate into astroglial cells by appropriate extracellular signals. PMID- 14745008 TI - Release of full-length 55-kDa TNF receptor 1 in exosome-like vesicles: a mechanism for generation of soluble cytokine receptors. AB - Soluble tumor necrosis factor receptors (TNFRs) are important modulators of TNF bioactivity. Proteolytic cleavage of the 28-kDa ectodomain of TNFR1 has been recognized as the mechanism by which soluble TNFR is shed. We now describe the release of exosome-like vesicles as a mechanism for the generation of soluble, full-length 55-kDa TNFR1. We found unexpectedly that the predominant form of soluble TNFR1 in human serum and lung epithelial lining fluid is a full-length 55 kDa protein. Furthermore, supernatants from human vascular endothelial cells contain only full-length 55-kDa TNFR1 that can be sedimented by high-speed centrifugation, floated on sucrose gradients at a density of 1.1 g/ml, and associated with vesicles that range in diameter from 20 nm to 50 nm. We conclude that the release of TNFR1 exosome-like vesicles represents a previously unrecognized mechanism by which constitutive production of soluble cytokine receptors may be regulated, independent of ectodomain cleavage by receptor sheddases. PMID- 14745009 TI - Functional reclassification of the putative cinnamyl alcohol dehydrogenase multigene family in Arabidopsis. AB - Of 17 genes annotated in the Arabidopsis genome database as cinnamyl alcohol dehydrogenase (CAD) homologues, an in silico analysis revealed that 8 genes were misannotated. Of the remaining nine, six were catalytically competent for NADPH dependent reduction of p-coumaryl, caffeyl, coniferyl, 5-hydroxyconiferyl, and sinapyl aldehydes, whereas three displayed very low activity and only at very high substrate concentrations. Of the nine putative CADs, two (AtCAD5 and AtCAD4) had the highest activity and homology (approximately 83% similarity) relative to bona fide CADs from other species. AtCAD5 used all five substrates effectively, whereas AtCAD4 (of lower overall catalytic capacity) poorly used sinapyl aldehyde; the corresponding 270-fold decrease in k(enz) resulted from higher K(m) and lower k(cat) values, respectively. No CAD homologue displayed a specific requirement for sinapyl aldehyde, which was in direct contrast with unfounded claims for a so-called sinapyl alcohol dehydrogenase in angiosperms. AtCAD2, 3, as well as AtCAD7 and 8 (highest homology to sinapyl alcohol dehydrogenase) were catalytically less active overall by at least an order of magnitude, due to increased K(m) and lower k(cat) values. Accordingly, alternative and/or bifunctional metabolic roles of these proteins in plant defense cannot be ruled out. Comprehensive analyses of lignified tissues of various Arabidopsis knockout mutants (for AtCAD5, 6, and 9) at different stages of growth/development indicated the presence of functionally redundant CAD metabolic networks. Moreover, disruption of AtCAD5 expression had only a small effect on either overall lignin amounts deposited, or on syringyl-guaiacyl compositions, despite being the most catalytically active form in vitro. PMID- 14745010 TI - Neanderthal taxonomy reconsidered: implications of 3D primate models of intra- and interspecific differences. AB - The taxonomic status of Neanderthals lies at the center of the modern human origins debate. Proponents of the single-origin model often view this group as a distinct species with little or no contribution to the evolution of modern humans. Adherents to the regional continuity model consider Neanderthals a subspecies or population of Homo sapiens, which contributed significantly to the evolution of early modern Europeans. Paleontologists generally agree that fossil species should be equivalent to extant ones in the amount of their morphological variation. Recognition of fossil species therefore hinges on analogy to living species. A previous study by one of the authors and recent work by other researchers [Schillachi, M. A. & Froelich, J. W. (2001) Am. J. Phys. Anthropol. 115, 157-166] have supported specific status for Neanderthals based on analogy to chimpanzees and Sulawesi macaques, respectively. However, these taxa may not be the most appropriate models for Pleistocene humans. Here we test the hypothesis that Neanderthals represent a subspecies of H. sapiens by comparing the degree of their morphological differentiation from modern humans to that found within and between 12 species of extant primates. The model taxa comprised >1,000 specimens, including phylogenetic (modern humans and African apes) and ecological (eight papionin taxa) models for Pleistocene humans. Morphological distances between model taxon pairs were compared to the distances between Neanderthals and modern humans obtained by using a randomization technique. Results strongly support a specific distinction for Neanderthals. PMID- 14745011 TI - The 1.8-A resolution crystal structure of YDR533Cp from Saccharomyces cerevisiae: a member of the DJ-1/ThiJ/PfpI superfamily. AB - The yeast gene YDR533C encodes a protein belonging to the DJ-1/ThiJ/PfpI superfamily. This family includes the human protein DJ-1, which is mutated in autosomal recessive early-onset Parkinson's disease. The function of DJ-1 and its yeast homologue YDR533Cp is unknown. We report here the crystal structure of YDR533Cp at 1.8-A resolution. The structure indicates that the closest relative to YDR533Cp is the Escherichia coli heat shock protein Hsp31 (YedU), which has both chaperone and protease activity. As expected, the overall fold of the core domain of YDR533Cp is also similar to that of DJ-1 and the bacterial protease PfpI. YDR533Cp contains a possible catalytic triad analogous to that of Hsp31 and an additional domain that is present in Hsp31 but is not seen in DJ-1 and other members of the family. The cysteine in this triad (Cys-138) is oxidized in this crystal structure, similar to modifications seen in the corresponding cysteine in the crystal structure of DJ-1. YDR533Cp appears to be a dimer both in solution and the crystal, but this dimer is formed by a different interface than that found in Hsp31 or other members of the superfamily. PMID- 14745012 TI - A Sertoli cell-selective knockout of the androgen receptor causes spermatogenic arrest in meiosis. AB - Androgens control spermatogenesis, but germ cells themselves do not express a functional androgen receptor (AR). Androgen regulation is thought to be mediated by Sertoli and peritubular myoid cells, but their relative roles and the mechanisms involved remain largely unknown. Using Cre/loxP technology, we have generated mice with a ubiquitous knockout of the AR as well as mice with a selective AR knockout in Sertoli cells (SC) only. Mice with a floxed exon 2 of the AR gene were crossed with mice expressing Cre recombinase ubiquitously or selectively in SC (under control of the anti-Mullerian hormone gene promoter). AR knockout males displayed a complete androgen insensitivity phenotype. Testes were located abdominally, and germ cell development was severely disrupted. In contrast, SC AR knockout males showed normal testis descent and development of the male urogenital tract. Expression of the homeobox gene Pem, which is androgen regulated in SC, was severely decreased. Testis weight was reduced to 28% of that in WT littermates. Stereological analysis indicated that the number of SC was unchanged, whereas numbers of spermatocytes, round spermatids, and elongated spermatids were reduced to 64%, 3%, and 0% respectively of WT. These changes were associated with increased germ cell apoptosis and grossly reduced expression of genes specific for late spermatocyte or spermatid development. It is concluded that cell-autonomous action of the AR in SC is an absolute requirement for androgen maintenance of complete spermatogenesis, and that spermatocyte/spermatid development/survival critically depends on androgens. PMID- 14745013 TI - The Drosophila gene Start1: a putative cholesterol transporter and key regulator of ecdysteroid synthesis. AB - Human metastatic lymph node 64 (MLN64) is a transmembrane protein that shares homology with the cholesterol-binding vertebrate steroid acute regulatory protein (StAR)-related lipid transfer domain (START) and is involved in cholesterol traffic and steroid synthesis. We identified a Drosophila melanogaster gene whose putative protein product shows extensive homology with MLN64 and that we name Start1 (FlyBase CG3522). The putative Start1 protein, derived from Start1 cDNA sequences, contains an additional 122 aa of unknown function within the StAR related lipid transfer domain. Similar inserts seem to exist in the Start1 homologues of Drosophila pseudoobscura and Anopheles gambiae, but not in the homologous protein of the urochordate Ciona intestinalis. Immunostaining using an insert-specific antibody confirms the presence of the insert in the cytoplasm. Whereas RT-PCR data indicate that Start1 is expressed ubiquitously, RNA in situ hybridizations demonstrate its overexpression in prothoracic gland cells, where ecdysteroids are synthesized from cholesterol. Transcripts of Start1 are detectable in embryonic ring gland progenitor cells and are abundant in prothoracic glands of larvae showing wave-like expression during larval stages. In adults, Start1 is expressed in nurse cells of the ovary. These observations are consistent with the assumption that Start1 plays a key role in the regulation of ecdysteroid synthesis. Vice versa, the expression of Start1 itself seems to depend on ecdysone, as in the ecdysone-deficient mutant ecd-1, Start1 expression is severely reduced. PMID- 14745015 TI - Method for analyzing signaling networks in complex cellular systems. AB - Now that the human genome has been sequenced, the challenge of assigning function to human genes has become acute. Existing approaches using microarrays or proteomics frequently generate very large volumes of data not directly related to biological function, making interpretation difficult. Here, we describe a technique for integrative systems biology in which: (i) primary cells are cultured under biologically meaningful conditions; (ii) a limited number of biologically meaningful readouts are measured; and (iii) the results obtained under several different conditions are combined for analysis. Studies of human endothelial cells overexpressing different signaling molecules under multiple inflammatory conditions show that this system can capture a remarkable range of functions by a relatively small number of simple measurements. In particular, measurement of seven different protein levels by ELISA under four different conditions is capable of reconstructing pathway associations of 25 different proteins representing four known signaling pathways, implicating additional participants in the NF-kappaBorRAS/mitogen-activated protein kinase pathways and defining additional interactions between these pathways. PMID- 14745014 TI - Methionine sulfoxide reductase A protects neuronal cells against brief hypoxia/reoxygenation. AB - Hypoxia/reoxygenation induces cellular injury by promoting oxidative stress. Reversible oxidation of methionine in proteins involving the enzyme peptide methionine sulfoxide reductase type A (MSRA) is postulated to serve a general antioxidant role. Therefore, we examined whether overexpression of MSRA protected cells from hypoxia/reoxygenation injury. Brief hypoxia increased the intracellular reactive oxygen species (ROS) level in PC12 cells and promoted apoptotic cell death. Adenovirus-mediated overexpression of MSRA significantly diminished the hypoxia-induced increase in ROS and facilitated cell survival. Measurements of the membrane potentials of intact mitochondria in PC12 cells and of isolated rat liver mitochondria showed that hypoxia induced depolarization of the mitochondrial membrane. The results demonstrate that MSRA plays a protective role against hypoxia/reoxygenation-induced cell injury and suggest the therapeutic potential of MSRA in ischemic heart and brain disease. PMID- 14745016 TI - T cell receptor zeta reconstitution fails to restore responses of T cells rendered hyporesponsive by tumor necrosis factor alpha. AB - Expression and function of the antigen T cell receptor (TCR) play a central role in regulating immune responsiveness. Accordingly, targeting the expression of TCRalphabeta or its associated CD3 subunits profoundly influences T cell development and adaptive immunity. Down-regulation of the invariant TCRzeta chain has been documented in a wide variety of chronic inflammatory and infectious diseases, and is thought to contribute to the paradoxical immune suppression observed in these diseases. Previously, we reported that prolonged exposure of T cell hybridoma clones to tumor necrosis factor alpha (TNF) induces nondeletional and reversible hyporesponsiveness to TCR engagement, associated with down regulation of TCRzeta chain expression, impaired TCR/CD3 complex assembly, and attenuation of TCR-induced membrane proximal tyrosine phosphorylation. Here, we have tested whether receptor specific T cell responses are rescued in TNF-treated T cell hybridomas by retroviral-mediated expression of zeta-chimeric (C2zeta) receptors or wild-type TCRzeta. Expression of C2zeta receptors at the cell surface is relatively refractory to chronic TNF stimulation. However, C2zeta receptor function depends on association with endogenous TCRzeta chains, whose expression is down-regulated by TNF, and so C2 receptor specific responses are attenuated in TNF-treated T cells. Unexpectedly, overexpression of wild-type TCRzeta maintains cell surface TCR/CD3 complex expression but fails to rescue receptor proximal signaling in TNF-treated T cells, suggesting the existence of hitherto unrecognized mechanisms through which TNF regulates T cell responsiveness. We provide additional evidence that TNF also uncouples distal TCR signaling pathways independently of its effects on TCRzeta expression. PMID- 14745017 TI - Interferon antagonist proteins of influenza and vaccinia viruses are suppressors of RNA silencing. AB - Homology-dependent RNA silencing occurs in many eukaryotic cells. We reported recently that nodaviral infection triggers an RNA silencing-based antiviral response (RSAR) in Drosophila, which is capable of a rapid virus clearance in the absence of expression of a virus-encoded suppressor. Here, we present further evidence to show that the Drosophila RSAR is mediated by the RNA interference (RNAi) pathway, as the viral suppressor of RSAR inhibits experimental RNAi initiated by exogenous double-stranded RNA and RSAR requires the RNAi machinery. We demonstrate that RNAi also functions as a natural antiviral immunity in mosquito cells. We further show that vaccinia virus and human influenza A, B, and C viruses each encode an essential protein that suppresses RSAR in Drosophila. The vaccinia and influenza viral suppressors, E3L and NS1, are distinct double stranded RNA-binding proteins and essential for pathogenesis by inhibiting the mammalian IFN-regulated innate antiviral response. We found that the double stranded RNA-binding domain of NS1, implicated in innate immunity suppression, is both essential and sufficient for RSAR suppression. These findings provide evidence that mammalian virus proteins can inhibit RNA silencing, implicating this mechanism as a nucleic acid-based antiviral immunity in mammalian cells. PMID- 14745018 TI - Estrogen receptor beta inhibits 17beta-estradiol-stimulated proliferation of the breast cancer cell line T47D. AB - Estrogen receptor (ER) beta counteracts the activity of ERalpha in many systems. In agreement with this, we show in this study that induced expression of ERbeta in the breast cancer cell line T47D reduces 17beta-estradiol-stimulated proliferation when expression of ERbeta mRNA equals that of ERalpha. Induction of ERbeta reduces growth of exponentially proliferating cells with a concomitant decrease in components of the cell cycle associated with proliferation, namely cyclin E, Cdc25A (a key regulator of Cdk2), p45(Skp2) (a key regulator of p27(Kip1) proteolysis), and an increase in the Cdk inhibitor p27(Kip1). We also observed a reduced Cdk2 activity. These findings suggest a possible role for ERbeta in breast cancer and imply that ERbeta-specific ligands may reduce proliferation of ER-positive breast cancer cells through actions on the G(1) phase cell-cycle machinery. PMID- 14745019 TI - Folate synthesis in plants: the p-aminobenzoate branch is initiated by a bifunctional PabA-PabB protein that is targeted to plastids. AB - It is not known how plants synthesize the p-aminobenzoate (PABA) moiety of folates. In Escherichia coli, PABA is made from chorismate in two steps. First, the PabA and PabB proteins interact to catalyze transfer of the amide nitrogen of glutamine to chorismate, forming 4-amino-4-deoxychorismate (ADC). The PabC protein then mediates elimination of pyruvate and aromatization to give PABA. Fungi, actinomycetes, and Plasmodium spp. also synthesize PABA but have proteins comprising fused domains homologous to PabA and PabB. These bipartite proteins are commonly called "PABA synthases," although it is unclear whether they produce PABA or ADC. Genomic approaches identified Arabidopsis and tomato cDNAs encoding bipartite proteins containing fused PabA and PabB domains, plus a putative chloroplast targeting peptide. These cDNAs encode functional enzymes, as demonstrated by complementation of an E. coli pabA pabB double mutant and a yeast PABA-synthase deletant. The partially purified recombinant Arabidopsis protein did not produce PABA unless the E. coli PabC enzyme was added, indicating that it forms ADC, not PABA. The enzyme behaved as a monomer in size-exclusion chromatography and was not inhibited by physiological concentrations of PABA, its glucose ester, or folates. When the putative targeting peptide was fused to GFP and expressed in protoplasts, the fusion protein appeared only in chloroplasts, indicating that PABA synthesis is plastidial. In the pericarp of tomato fruit, the PabA-PabB mRNA level fell drastically as ripening advanced, but there was no fall in total PABA content, which stayed between 0.7 and 2.3 nmol.g(-1) fresh weight. PMID- 14745021 TI - Protein kinases talk to lipid phosphatases at the synapse. PMID- 14745020 TI - Avian influenza A virus (H7N7) associated with human conjunctivitis and a fatal case of acute respiratory distress syndrome. AB - Highly pathogenic avian influenza A viruses of subtypes H5 and H7 are the causative agents of fowl plague in poultry. Influenza A viruses of subtype H5N1 also caused severe respiratory disease in humans in Hong Kong in 1997 and 2003, including at least seven fatal cases, posing a serious human pandemic threat. Between the end of February and the end of May 2003, a fowl plague outbreak occurred in The Netherlands. A highly pathogenic avian influenza A virus of subtype H7N7, closely related to low pathogenic virus isolates obtained from wild ducks, was isolated from chickens. The same virus was detected subsequently in 86 humans who handled affected poultry and in three of their family members. Of these 89 patients, 78 presented with conjunctivitis, 5 presented with conjunctivitis and influenza-like illness, 2 presented with influenza-like illness, and 4 did not fit the case definitions. Influenza-like illnesses were generally mild, but a fatal case of pneumonia in combination with acute respiratory distress syndrome occurred also. Most virus isolates obtained from humans, including probable secondary cases, had not accumulated significant mutations. However, the virus isolated from the fatal case displayed 14 amino acid substitutions, some of which may be associated with enhanced disease in this case. Because H7N7 viruses have caused disease in mammals, including horses, seals, and humans, on several occasions in the past, they may be unusual in their zoonotic potential and, thus, form a pandemic threat to humans. PMID- 14745022 TI - An ultradian clock shapes genome expression in yeast. PMID- 14745023 TI - Increase of the RNA-binding protein HuD and posttranscriptional up-regulation of the GAP-43 gene during spatial memory. AB - Neuronal ELAV-like proteins (HuB, HuC, and HuD) are highly conserved RNA-binding proteins able to selectively associate with the 3' UTR of a subset of target mRNAs and increase their cytoplasmic stability and rate of translation. We previously demonstrated the involvement of these proteins in learning, reporting that they undergo a sustained up-regulation in the hippocampus of mice trained in a spatial discrimination task. Here, we extend this finding, showing that a similar up-regulation occurs in the hippocampus of rats trained in another spatial learning paradigm, the Morris water maze. HuD, a strictly neuron-specific ELAV-like protein, is shown to increase after learning, with a preferential binding to the cytoskeletal fraction. HuD up-regulation is associated with an enhancement of GAP-43 mRNA and protein levels, with an apparently increased HuD colocalization with the GAP-43 mRNA and an increased association of neuronal ELAV like proteins with the GAP-43 mRNA. These learning-dependent biochemical events appear to be spatiotemporally controlled, because they do not occur in another brain region involved in learning, the retrosplenial cortex, and at the level of protein expression they show extinction 1 month after training despite memory retention. By contrast, HuD mRNA levels still remain increased after 1 month in the CA1 region. This persistence may have implications for long-term memory recall. PMID- 14745024 TI - Production of homoplasmic xenomitochondrial mice. AB - The unique features of mtDNA, together with the lack of a wide range of mouse cell mtDNA mutants, have hampered the creation of mtDNA mutant mice. To overcome these barriers mitochondrial defects were created by introducing mitochondria from different mouse species into Mus musculus domesticus (Mm) mtDNA-less (rho(0)) L cells. Introduction of the closely related Mus spretus (Ms) or the more divergent Mus dunni (Md) mitochondria resulted in xenocybrids exhibiting grossly normal respiratory function, but mild metabolic deficiencies, with 2- and 2.5-fold increases in lactate production compared with controls. The transfer of this model from in vitro to in vivo studies was achieved by introducing Ms and Md mitochondria into rhodamine-6G-treated Mm mouse embryonic stem (ES) cells. The resultant xenocybrid ES cells remained pluripotent, and live-born chimerae were produced from both Ms and Md xenocybrid ES cells. Founder chimeric females (G(0)) were mated with successful germ-line transmission of Ms or Md mtDNA to homoplasmic G(1) offspring. These xenocybrid models represent the first viable transmitochondrial mice with homoplasmic replacement of endogenous mtDNA and confirm the feasibility of producing mitochondrial defects in mice by using a xenomitochondrial approach. PMID- 14745025 TI - Stem cell-mediated muscle regeneration is enhanced by local isoform of insulin like growth factor 1. AB - We investigated the mechanism whereby expression of a transgene encoding a locally acting isoform of insulin-like growth factor 1 (mIGF-1) enhances repair of skeletal muscle damage. Increased recruitment of proliferating bone marrow cells to injured MLC/mIgf-1 transgenic muscles was accompanied by elevated bone marrow stem cell production in response to distal trauma. Regenerating MLC/mIgf-1 transgenic muscles contained increased cell populations expressing stem cell markers, exhibited accelerated myogenic differentiation, expressed markers of regeneration and readily converted cocultured bone marrow to muscle. These data implicate mIGF-1 as a powerful enhancer of the regeneration response, mediating the recruitment of bone marrow cells to sites of tissue damage and augmenting local repair mechanisms. PMID- 14745027 TI - Evolution of the MAT locus and its Ho endonuclease in yeast species. AB - The genetics of the mating-type (MAT) locus have been studied extensively in Saccharomyces cerevisiae, but relatively little is known about how this complex system evolved. We compared the organization of MAT and mating-type-like (MTL) loci in nine species spanning the hemiascomycete phylogenetic tree. We inferred that the system evolved in a two-step process in which silent HMR/HML cassettes appeared, followed by acquisition of the Ho endonuclease from a mobile genetic element. Ho-mediated switching between an active MAT locus and silent cassettes exists only in the Saccharomyces sensu stricto group and their closest relatives: Candida glabrata, Kluyveromyces delphensis, and Saccharomyces castellii. We identified C. glabrata MTL1 as the ortholog of the MAT locus of K. delphensis and show that switching between C. glabrata MTL1a and MTL1alpha genotypes occurs in vivo. The more distantly related species Kluyveromyces lactis has silent cassettes but switches mating type without the aid of Ho endonuclease. Very distantly related species such as Candida albicans and Yarrowia lipolytica do not have silent cassettes. In Pichia angusta, a homothallic species, we found MATalpha2, MATalpha1, and MATa1 genes adjacent to each other on the same chromosome. Although some continuity in the chromosomal location of the MAT locus can be traced throughout hemiascomycete evolution and even to Neurospora, the gene content of the locus has changed with the loss of an HMG domain gene (MATa2) from the MATa idiomorph shortly after HO was recruited. PMID- 14745026 TI - Strong selective sweep associated with a transposon insertion in Drosophila simulans. AB - We know little about several important properties of beneficial mutations, including their mutational origin, their phenotypic effects (e.g., protein structure changes vs. regulatory changes), and the frequency and rapidity with which they become fixed in a population. One signature of the spread of beneficial mutations is the reduction of heterozygosity at linked sites. Here, we present population genetic data from several loci across chromosome arm 2R in Drosophila simulans. A 100-kb segment from a freely recombining region of this chromosome shows extremely reduced heterozygosity in a California population sample, yet typical levels of divergence between species, suggesting that at least one episode of strong directional selection has occurred in the region. The 5' flanking sequence of one gene in this region, Cyp6g1 (a cytochrome P450), is nearly fixed for a Doc transposable element insertion. Presence of the insertion is correlated with increased transcript abundance of Cyp6g1, a phenotype previously shown to be associated with insecticide resistance in Drosophila melanogaster. Surveys of nucleotide variation in the same genomic region in an African D. simulans population revealed no evidence for a high-frequency Doc element and no evidence for reduced polymorphism. These data are consistent with the notion that the Doc element is a geographically restricted beneficial mutation. Data from D. simulans Cyp6g1 are paralleled in many respects by data from its sister species D. melanogaster. PMID- 14745028 TI - Transgenic zebrafish produced by retroviral infection of in vitro-cultured sperm. AB - Transgenic modification of sperm before fertilization has distinct advantages over conventional transgenic methods. The primary advantage is that the mosaicism inherent in those other techniques is avoided. A culture system using primary cultures of zebrafish male germ cells, in which the differentiation from spermatogonia to functional sperm can occur in vitro, provides the opportunity for genetic modification of sperm in vitro. Here, we report the production of transgenic zebrafish from cultured sperm. The sperm were differentiated from premeiotic germ cells infected with a pseudotyped retrovirus in vitro. The collected sperm were used to perform successful in vitro fertilizations, and transgenic embryos were identified. The transgenic fish transmitted the proviral integration to the next generation in a Mendelian fashion. We report the generation of a transgenic animal by cultured sperm and open the door to many exciting possibilities for the rapid generation of transgenic lines in model organisms such as zebrafish or other animal systems that are otherwise intractable to transgenesis. PMID- 14745029 TI - An Arabidopsis indole-3-butyric acid-response mutant defective in PEROXIN6, an apparent ATPase implicated in peroxisomal function. AB - Genetic evidence suggests that plant peroxisomes are the site of fatty acid beta oxidation and conversion of the endogenous auxin indole-3-butyric acid (IBA) to the active hormone indole-3-acetic acid. Arabidopsis mutants that are IBA resistant and sucrose dependent during early development are likely to have defects in beta-oxidation of both IBA and fatty acids. Several of these mutants have lesions in peroxisomal protein genes. Here, we describe the Arabidopsis pex6 mutant, which is resistant to the inhibitory effects of IBA on root elongation and the stimulatory effects of IBA on lateral root formation. pex6 also is sucrose dependent during early seedling development and smaller and more pale green than WT throughout development. PEX6 encodes an apparent ATPase similar to yeast and human proteins required for peroxisomal biogenesis, and a human PEX6 cDNA can rescue the Arabidopsis pex6 mutant. The pex6 mutant has reduced levels of the peroxisomal matrix protein receptor PEX5, and pex6 defects can be partially rescued by PEX5 overexpression. These results suggest that PEX6 may facilitate PEX5 recycling and thereby promote peroxisomal matrix protein import. PMID- 14745030 TI - The rise and fall of a networked society: a formal model. AB - In a well networked community, there is intense social interaction, and information disseminates briskly and broadly. This is important if the environment is volatile (i.e., keeps changing) and individuals never stop searching for fresh opportunities. Here, we present a simple model that attributes the rise of a dynamic society to the emergence of some key features in its social network. We also explain the apparently paradoxical observation that although such features do not necessarily materialize even under favorable conditions they display a significant resilience to deteriorating conditions. We interpret these findings as a discontinuous phase transition in the network formation process. PMID- 14745031 TI - Induction of aneuploidy by increasing chromosomal instability during dedifferentiation of hepatocellular carcinoma. AB - To gain more insight into the role of chromosomal instability (CIN), the cytogenetic hallmark of most solid tumors, we performed fluorescence in situ hybridization (FISH) on interphase nuclei of cytological specimens enabling the correct detection of chromosome copies in intact tumor cells of 18 well (G1), moderately (G2), or poorly (G3) differentiated hepatocellular carcinomas (HCCs). A close correlation between the morphological dedifferentiation and increasing copy numbers and variation of FISH signals was seen for chromosomes 1 and 8, respectively (P < or = 0.0002). Four HCC G1 had constant chromosome patterns for chromosomes 1 and/or 8 with a mean of signals per nucleus < or =5.08 and < or =3 different signal combinations, indicating a low level of CIN, as confirmed by FISH using probes for centromeres of chromosomes 3, 7, and 17. In contrast to this, five HCC G2-3 revealed > or =8.46 signals per nucleus and 23-41 different signal combinations, indicating high levels of CIN. In the remaining cases, signal counts from 5.96-8.46 and 7-15 combinations were seen. Here, nuclei with constant aberration patterns and low copy numbers occurred alongside nuclei with inconstant patterns and high copy numbers. It is evident that in these cases a transition from well to moderately differentiated HCC developed in parallel to an increase in CIN, possibly induced by a major dysregulation of mitotic control mechanisms. In conclusion, CIN may induce a stepwise increase of aneuploidy in HCC that is mirrored by the morphological dedifferentiation of tumor cells. PMID- 14745032 TI - Control of late off-center cone bipolar cell differentiation and visual signaling by the homeobox gene Vsx1. AB - Retinal bipolar cells are interneurons that transmit visual signals from photoreceptors to ganglion cells. Although the visual pathways mediated by bipolar cells have been well characterized, the genes that regulate their development and function are largely unknown. To determine the role in bipolar cell development of the homeobox gene Vsx1, whose retinal expression is restricted to a major subset of differentiating and mature cone bipolar (CB) cells, we targeted the gene in mice. Bipolar cell fate was not altered in the absence of Vsx1 function, because the pan-bipolar markers Chx10 and Ret-B1 continued to be expressed in inner nuclear layer neurons labeled by the Vsx1 targeting reporter gene, tauLacZ. The specification, number, and gross morphology of the subset of on-center and off-center (OFF)-CB cells defined by tauLacZ expression from the Vsx1 locus were also normal in Vsx1(tauLacZ)/Vsx1(tauLacZ) mice. However, the terminal differentiation of OFF-CB cells in the retina of Vsx1(tauLacZ)/Vsx1(tauLacZ) mice was incomplete, as demonstrated by a substantial reduction in the expression of at least four markers (recoverin, NK3R, Neto1, and CaB5) for these interneurons. These molecular abnormalities were associated with defects in retinal function and documented by electroretinography and in vitro ganglion cell recordings specific to cone visual signaling. In particular, there was a general reduction in the light-mediated activity of OFF, but not on-center, ganglion cells. Thus, Vsx1 is required for the late differentiation and function of OFF-CB cells and is associated with a heritable OFF visual pathway-specific retinal defect. PMID- 14745033 TI - Costimulatory ligand 4-1BBL (CD137L) as an efficient adjuvant for human antiviral cytotoxic T cell responses. AB - Effective adjuvants capable of inducing strong cytotoxic T cell responses in humans are lacking. In this study, we tested 4-1BBL as an adjuvant for activation of human memory antiviral CD8 T cell responses ex vivo. A recombinant replication defective 4-1BBL adenovirus was used to convert autologous monocytes into efficient antigen-presenting cells after overnight incubation, bypassing the need to generate dendritic cells. Together with viral peptides, 4-1BBL led to robust memory responses of human Epstein-Barr virus- and influenza virus-specific cytotoxic T cells, with expansion of peptide-specific CD8 effector cells; up regulation of Bcl-x(L), granzyme A, and perforin; enhanced cytotoxic activity; and increased cytokine production. The response was significant even at a 100 fold lower peptide dose, compared with responses obtained with control adenovirus. Adenovirus-delivered B7.1 also expanded and activated virus-specific CD8 T cells, but 4-1BBL was more effective in driving the T cells toward a more fully differentiated CD27(-) effector state. Thus, 4-1BBL is a promising adjuvant for human memory CD8 T cells and will likely be most effective in the boost phase of a prime-boost strategy. PMID- 14745034 TI - Reorganization of the auditory cortex specialized for echo-delay processing in the mustached bat. AB - Focal excess sensory stimulation evokes reorganization of a sensory system. It is usually an expansion of the neural representation of that stimulus resulting from the shifts of the tuning curves (receptive fields) of neurons toward those of the stimulated neurons. The auditory cortex of the mustached bat has an area that is highly specialized for the processing of target-distance information carried by echo delays. In this area, however, reorganization is due to shifts of the delay tuning curves of neurons away from those of the stimulated cortical neurons. Elimination of inhibition in the target-distance processing area in the auditory cortex by a drug reverses the direction of the shifts in neural tuning curves. Therefore, such unique reorganization in the time domain is due to strong lateral inhibition in the highly specialized area of the auditory cortex. PMID- 14745035 TI - Protein phosphorylation can regulate metabolite concentrations rather than control flux: the example of glycogen synthase. AB - Despite dramatic increases in glucose influx during the transition from fasting to fed states, plasma glucose concentration remains tightly controlled. This constancy is in large part due to the capacity of skeletal muscle to absorb excess glucose and store it as glycogen. The magnitude of this capacity is controlled by insulin by way of regulated insertion of glucose transporters into the muscle cell membrane. Here, we examine the mechanism by which muscle cells are able to tolerate large flux increases across their transporters without significantly changing their own metabolite pools. MCA was used to probe data sets that measured the effects of changing plasma glucose and/or insulin concentrations on the rates of glycogen synthesis and the concentrations of metabolites, particularly glucose-6-phosphate. We find that homeostasis is achieved by insulin-dependent phosphorylation changes in GSase sensitivity to the upstream metabolite glucose-6-phosphate. The centrality of GSase to homeostasis resolves the paradox of its sensitivity to allosteric and covalent regulation despite its minimal role in flux control. The importance of this role for enzymatic phosphorylation to diabetes pathology is discussed, and its general applicability is suggested. PMID- 14745036 TI - Herpes simplex virus infections are arrested in Oct-1-deficient cells. AB - Expression of the herpes simplex virus (HSV) immediate early (IE) genes is regulated by a multiprotein complex that is assembled on the TAATGARAT enhancer core element. The complex contains the cellular POU domain protein Oct-1, the viral transactivator VP16, and the cellular cofactor host cell factor 1. The current model suggests that the assembly depends on recognition of the core element by Oct-1. Here, HSV infection of Oct-1-deficient mouse embryonic fibroblast cells demonstrates that Oct-1 is critical for IE gene expression at low multiplicities of infection (moi). However, the protein is not essential for IE gene expression at high moi, indicating that VP16-mediated transcriptional induction through other IE regulatory elements is also important. This induction depends, at least in part, on the GA-binding protein binding elements that are present in each IE enhancer domain. Surprisingly, whereas the viral IE genes are expressed after high moi infection of Oct-1-deficient cells, the assembly of viral replication factories is severely impaired, revealing a second critical role for Oct-1 in HSV replication. The results have implications for both the HSV lytic and latency-reactivation cycles. PMID- 14745037 TI - Ultrafast electron crystallography: transient structures of molecules, surfaces, and phase transitions. AB - The static structure of macromolecular assemblies can be mapped out with atomic scale resolution by using electron diffraction and microscopy of crystals. For transient nonequilibrium structures, which are critical to the understanding of dynamics and mechanisms, both spatial and temporal resolutions are required; the shortest scales of length (0.1-1 nm) and time (10(-13) to 10(-12) s) represent the quantum limit, the nonstatistical regime of rates. Here, we report the development of ultrafast electron crystallography for direct determination of structures with submonolayer sensitivity. In these experiments, we use crystalline silicon as a template for different adsorbates: hydrogen, chlorine, and trifluoroiodomethane. We observe the coherent restructuring of the surface layers with subangstrom displacement of atoms after the ultrafast heat impulse. This nonequilibrium dynamics, which is monitored in steps of 2 ps (total change 4 centuries ago, the legacy of these human disturbances is still evident in the pond's present-day limnology and is characterized by elevated nutrient concentrations and atypical biota. This is the earliest reported paleolimnological record of changes in aquatic ecology associated with local human activities in Canada or the United States, or for any circumpolar ecosystem. PMID- 14745044 TI - Disruption of Ixodes scapularis anticoagulation by using RNA interference. AB - Ixodes scapularis ticks transmit many pathogens, including Borrelia burgdorferi, Anaplasma phagocytophilum, and Babesia microti. Vaccines directed against arthropod proteins injected into the host during tick engorgement could prevent numerous infectious diseases. Salp14, a salivary anticoagulant, poses a key target for such intervention. Salp14 is the prototypic member of a family of potential I. scapularis anticoagulants, expressed and secreted in tick saliva during tick feeding. RNA interference was used to assess the role of Salp14 in tick feeding. Salp14 and its paralogs were silenced, as demonstrated by the reduction of mRNA and protein specific for these antigens. Tick salivary glands lacking Salp14 had reduced anticoagulant activity, as revealed by a 60-80% reduction of anti-factor Xa activity. Silencing the expression of salp14 and its paralogs also reduced the ability of I. scapularis to feed, as demonstrated by a 50-70% decline in the engorgement weights. Because ticks have several anticoagulants, it is likely that the expression of multiple anticoagulants in I. scapularis saliva would have to be ablated simultaneously to abolish tick feeding. These studies demonstrate that RNA interference can silence I. scapularis genes and disrupt their physiologic function in vivo, and they identify vaccine candidates that can alter vector engorgement. PMID- 14745045 TI - Suppressing immunity in advancing MS: too much too late, or too late for much? PMID- 14745046 TI - Testosterone and Alzheimer disease: is it men's turn now? PMID- 14745047 TI - Back to school on APS: rethinking the risk of recurrent stroke in children. PMID- 14745048 TI - Epilepsy versus antiepileptic drugs and gonadal function in men. PMID- 14745050 TI - Ophthalmologic features of Parkinson's disease. AB - Patients with Parkinson's disease (PD) commonly complain of impaired visual function and difficulty reading, despite normal visual acuity. Although previous studies have evaluated contrast sensitivity, color vision, visuospatial processing, visual hallucinations, and ocular movements, none has systematically evaluated the ocular complaints and ocular findings of PD patients. Thirty patients with early untreated PD and 31 control subjects without neurologic or known ocular diseases were ophthalmologically evaluated for the frequency of visual complaints, dry eyes, blepharitis, visual hallucinations, reduced blink rate, blepharospasm, and convergence insufficiency. Ocular complaints suggesting ocular surface irritation, altered tear film, visual hallucinations, blepharospasm, decreased blink rate, and decreased convergence amplitudes were more common in PD patients than in control subjects. These findings likely account for many of the visual difficulties commonly encountered by PD patients. These ocular abnormalities frequently respond to treatment. PMID- 14745051 TI - DLB fluctuations: specific features that reliably differentiate DLB from AD and normal aging. AB - OBJECTIVE: To determine whether certain aspects of fluctuations reliably distinguish dementia with Lewy bodies (DLB) from Alzheimer's disease (AD) and normal aging. METHODS: Participants included 200 community-dwelling cognitively normal elderly persons, 70 DLB patients, and 70 AD patients with collateral informants. A 19-item questionnaire was administered to the informants that queried about symptoms of fluctuations and delirium. RESULTS: Fluctuations occur infrequently in nondemented elderly persons aged 58 to 98 years. In contrast, four characteristics of fluctuations were found to significantly differentiate AD from DLB. These composite features include daytime drowsiness and lethargy, daytime sleep of 2 or more hours, staring into space for long periods, and episodes of disorganized speech. The presence of three or four features of this composite occurred in 63% of DLB patients compared with 12% of AD patients and 0.5% of normal elderly persons. Informant endorsement of three or four of these items yielded a positive predictive value of 83% for the clinical diagnosis of DLB against an alternate diagnosis of AD. Endorsement of fewer than three items had a negative predictive value of 70% for the absence of a clinical diagnosis of DLB in favor of AD. The authors present evidence of test-retest reliability, convergent validity, and empirical verification with a separate cross-validation sample. Fluctuations were not associated with any particular combination of hallucinations, parkinsonism, or REM sleep behavior disorder. CONCLUSIONS: Based on informant report, disturbed arousal and disorganized speech are specific aspects of fluctuations in dementia with Lewy bodies that reliably distinguish dementia with Lewy bodies from Alzheimer's disease and normal aging. PMID- 14745052 TI - Free testosterone and risk for Alzheimer disease in older men. AB - OBJECTIVE: To investigate the relationships between age-associated decreases in endogenous serum total testosterone (T) and a free T index (FTI) in men and the subsequent development of Alzheimer disease (AD). METHOD: The authors used a prospective, longitudinal design with follow-up in men since 1958. Participants were from the Baltimore Longitudinal Study of Aging, a community-dwelling volunteer sample with baseline ages of 32 to 87 years. All subjects were free of AD at baseline T assessment. Five hundred seventy-four men assessed at multiple time points were followed for a mean of 19.1 years (range, 4 to 37 years). Diagnoses of AD were based on biennial physical, neurologic, and neuropsychological evaluations. RESULTS: Diagnosis of AD was associated inversely with FTI by itself and after adjustments for age, education, smoking status, body mass index, diabetes, any cancer diagnoses, and hormone supplements. In separate analyses, total T and sex hormone binding globulin were not significant predictors after adjustment with covariates. Increases in the FTI were associated with decreased risk of AD (hazard ratio = 0.74; 95% CI = 0.57 to 0.96), a 26% decrease for each 10-nmol/nmol FTI increase. CONCLUSIONS: Calculated free testosterone concentrations were lower in men who developed Alzheimer disease, and this difference occurred before diagnosis. Future research may determine whether higher endogenous free testosterone levels offer protection against a diagnosis of Alzheimer disease in older men. PMID- 14745053 TI - Increased anticardiolipin antibody IgG titers do not predict recurrent stroke or TIA in children. AB - BACKGROUND: Increased anticardiolipin antibody (ACLA) immunoglobulin (Ig) G titers are commonly found in children with arterial ischemic stroke (AIS) or TIA (AIS/TIA). The associated risk of recurrent thromboembolism is unknown. OBJECTIVE: To determine the risk of recurrent thromboembolism associated with persistently increased ACLA titers of the IgG isotype in children with AIS/TIA. METHODS: The authors studied a cohort of children surviving first AIS/TIA tested by standardized ELISA for beta2-glycoprotein I-dependent ACLA of the IgG isotype. Children with ACLA titers >15 IgG phospholipid (GPL) units (per manufacturer's cutoff point) on more than two occasions > or =6 weeks apart were classified as ACLA-positive (ACLA+) and compared with ACLA-negative (ACLA-) children with respect to recurrent thromboembolic events (AIS/TIA, sinovenous thrombosis, and extracerebral thromboembolism). RESULTS: The authors recruited 34 ACLA+ children and 151 ACLA- children. Most ACLA+ children (30/34; 88%) had ACLA titers < or =40 GPL units. During the follow-up period (median duration, 2.8 years for ACLA+ children and 3.0 years for ACLA- children), AIS/TIA recurred in 26% of ACLA+ children and in 38% of ACLA- children; none developed sinovenous thrombosis or extracerebral thromboembolism. Based on survival analysis, this difference was nonsignificant (p = 0.54). Using binary partition evaluation, no titer criteria for ACLA positivity (range, 0 to 60 GPL units) predicted recurrent AIS/TIA. CONCLUSION: In children surviving arterial ischemic stroke/TIA, increased anticardiolipin antibody immunoglobulin G titers do not predict recurrent thromboembolism. PMID- 14745055 TI - High concordance for essential tremor in monozygotic twins of old age. AB - OBJECTIVE: To assess the relative contribution of genetic and environmental factors for the etiology of essential tremor (ET) and to explore the effect of different diagnostic criteria. METHODS: A total of 2,448 twins of the Danish twin registry aged 70 years or more were screened for ET by an interview and an Archimedes spiral test. All twin pairs (n = 162) with a positive screening test of at least one of the twins were recontacted and 218 individuals (109 pairs) were interviewed and examined by a movement disorder specialist. The consensus criteria of the Tremor Investigation Group were applied to diagnose ET. RESULTS: Twenty-nine twins fulfilled the criteria of definite, 7 of probable, and 56 of possible ET. The probandwise concordance rate for the broadest definition of ET was 77% for monozygotic twins (MZ) and 59% for dizygotic twins (DZ). However, in an analysis restricted to cases of probable and definite ET, the concordance rates were 93% and 29%. The heritability for the liability to ET ranged from 93% to 99% using a general population prevalence of 1.2% for white 70+-year-olds. The inclusion of probable and exclusion of possible cases in the diagnosis of ET produced the highest concordance rates. CONCLUSION: The high concordance among MZ twins of very old age in this first population-based twin study of ET suggests that a disease phenotype consisting of definite and probable ET has a high heritability and hence is a good candidate for a phenotype to be used in linkage studies. PMID- 14745054 TI - Unilateral pallidotomy versus bilateral subthalamic nucleus stimulation in PD: a randomized trial. AB - OBJECTIVE: To compare the efficacy of unilateral pallidotomy and bilateral subthalamic nucleus (STN) stimulation in patients with advanced Parkinson disease (PD) in a randomized, observer-blind, multicenter trial. METHODS: Thirty-four patients with advanced PD were randomly assigned to have unilateral pallidotomy or bilateral STN stimulation. The primary outcome was the change from baseline to 6 months in the motor part of the Unified PD Rating Scale (motor UPDRS) in the off phase. Secondary outcomes were parkinsonian symptoms in the on phase (motor UPDRS), dyskinesias (Clinical Dyskinesia Rating Scale and dyskinesias UPDRS), functional status (activities of daily living UPDRS and Schwab and England scale), PD Quality of Life questionnaire, changes in drug treatment, and adverse effects. RESULTS: The off phase motor UPDRS score improved from 46.5 to 37 points in the group of pallidotomy patients and from 51.5 to 26.5 in the STN stimulation patients (p = 0.002). Of the secondary outcome measures, on phase motor UPDRS and dyskinesias UPDRS improved significantly in favor of the STN stimulation patients. Reduction of antiparkinsonian drugs was greater after STN stimulation than after pallidotomy. One patient in each group had a major adverse effect. CONCLUSIONS: Bilateral STN stimulation is more effective than unilateral pallidotomy in reducing parkinsonian symptoms in patients with advanced PD. PMID- 14745056 TI - Painful stimuli evoke itch in patients with chronic pruritus: central sensitization for itch. AB - BACKGROUND: Central sensitization for pain is important for patients with chronic pain. The authors investigated a possible role of central sensitization for itch in patients with chronic pruritus. METHODS: Noxious stimuli were applied in lesional and visually nonlesional skin areas of 25 patients with atopic dermatitis, in lesional skin areas of 9 patients with psoriasis vulgaris, and in 20 healthy subjects. The stimuli included mechanical pinpricks, electrical stimuli, contact heat, and injection of low-pH solution. Intensities of itch and pain were assessed separately on a numeric rating scale. RESULTS: All the noxious stimuli primarily evoked pain in control subjects and patients with psoriasis vulgaris. In patients with atopic dermatitis, however, itch was evoked instead of burning pain. In their lesional skin, itch was the predominant sensation. Chemical stimuli evoked intense itch in lesional and visually healthy skin areas (the area under the curve of itch rating compared with the control, mean +/- SEM, 668 +/- 166 and 625 +/- 192 vs 38 +/- 23; p < 0.001; p < 0.01). Chemically induced itch also was observed in healthy subjects after a conditioning histamine stimulus of 15 minutes, but not after a conditioning histamine stimulus of 2 minutes. CONCLUSION: The chronic barrage of pruriceptive input may elicit central sensitization for itch so that nociceptive input no longer inhibits itch but on the contrary is perceived as itch. In contrast to the well-known A-fiber-mediated alloknesis and hyperknesis, this type of central sensitization appears to be elicited by C-nociceptors. PMID- 14745057 TI - Systemic lidocaine in pain due to peripheral nerve injury and predictors of response. AB - OBJECTIVE: To investigate the effects of IV lidocaine on spontaneous and evoked pain (allodynia and hyperalgesia) due to peripheral nerve injury (postherpetic neuralgia or nerve trauma) using quantitative sensory testing. METHOD: The authors randomized 22 patients to receive lidocaine 5 mg/kg IV during 30 minutes or placebo in a double-blind crossover design and 16 patients subsequently received mexiletine on an open basis titrated from 400 to 1,000 mg per day (mean 737 mg/day). RESULTS: Lidocaine induced a significant decrease in ongoing pain for up to 6 hours with a peak effect 60 to 120 minutes postinjection. The drug also decreased mechanical dynamic allodynia and static (punctate) mechanical allodynia/hyperalgesia, but not thermal allodynia and hyperalgesia. The effects of lidocaine and mexiletine on spontaneous pain intensity were significantly higher in patients with concomitant mechanical allodynia in comparison with those without allodynia. CONCLUSIONS: These data indicate modality-specific antihyperalgesic effects of IV lidocaine in patients with peripheral nerve injury. Patients with mechanical allodynia may be good candidates for treatment with local anesthetic-like drugs and possibly with other sodium-channel blockers. PMID- 14745058 TI - Spinal cord abnormalities in recently diagnosed MS patients: added value of spinal MRI examination. AB - OBJECTIVE: The most recent diagnostic criteria for multiple sclerosis (MS) ascertain that findings from spinal cord MRI can be used to demonstrate dissemination in space. Because little is known about the prevalence and characteristics of cord lesions early in the disease, the authors studied the prevalence of spinal cord abnormalities in patients with early-stage MS and assessed their impact on diagnostic classification. METHODS: The brains and spinal cords of 104 recently diagnosed patients with MS were examined. Median interval between first symptom and diagnosis was 18.4 months. The brain MRI protocol included before and after gadolinium axial T1-weighted conventional spin echo sequences and dual-echo spin-echo images. For spinal cord MRI, sagittal cardiac-triggered dual-echo T2-weighted and sagittal T1-weighted spin-echo images were included. Clinical assessment for each patient included age, sex, clinical signs for spinal cord involvement, and Expanded Disability Status Scale. RESULTS: Abnormal cord MRIs were found in 83% of patients, usually with only focal lesions. Diffuse cord abnormalities were found in 13% of patients, although in isolation they were found in only three patients. Focal cord lesions were often multiple (median number, 3.0), small (median, 0.8 vertebral segments), and primarily (56.4%) situated in the cervical spinal cord. In 68 of 104 patients (65.4%), two or more focal lesions were visible on spinal cord images. The criteria for dissemination in space, as defined in the McDonald criteria for the brain, were met in only 66.3% of the patients. This percentage increased to 84.6% when spinal cord MRI abnormalities were also included. CONCLUSION: Spinal cord abnormalities are prevalent in patients with early-stage MS, have distinct morphologic characteristics, and help to determine dissemination in space at time of diagnosis. PMID- 14745059 TI - Brain activation patterns associated with working memory in relapsing-remitting MS. AB - BACKGROUND: Patients with multiple sclerosis (MS) show changes in brain activation patterns during visual and motor tasks that include decreases in the typical local network for a function and increases in other brain regions. OBJECTIVE: To determine whether brain activation patterns associated with working memory are affected by MS. METHODS: Activation of working memory circuitry was examined using an fMRI n-back task in adults with mild relapsing-remitting MS (RRMS; n = 10) and demographically matched healthy controls (n = 10). RESULTS: Group differences in brain activation emerged during both low- and high-demand conditions (p < 0.001). Overall, patients showed less activation than controls in core prefrontal and parietal regions of working memory circuitry, and greater activation in other regions within and beyond typical working memory circuitry, including bilateral medial frontal, cingulate, parietal, bilateral middle temporal, and occipital regions. CONCLUSIONS: Relative to controls, patients with mild RRMS showed shifts in brain activation patterns within and beyond typical components of working memory circuitry. PMID- 14745060 TI - Peripheral blood leukocyte NO production in MS patients with a benign vs progressive course. AB - BACKGROUND: Nitric oxide (NO) may play a role in tissue destruction and axonal degeneration in multiple sclerosis (MS). OBJECTIVE: To investigate NO production by peripheral blood leukocytes (PBL) in patients with a benign and progressive course of MS. METHODS: PBL were isolated from 25 patients with a benign course of MS (BMS), 33 with secondary progressive MS (SPMS), 21 with primary progressive MS (PPMS), and 29 healthy individuals. Leukocyte supernatants were assayed for nitrite concentration, which is an index of NO generation, using the Griess reaction. Serum levels of tumor necrosis factor (TNF)alpha and interleukin (IL) 12 were measured using ELISA. RESULTS: Compared to healthy controls, nitrite concentrations were higher in patients with BMS (p < 0.001), SPMS (p < 0.001), and PPMS (p < 0.05). There were no significant differences among the three clinical subgroups of MS. There was a correlation between nitrite concentrations and serum levels of IL-12 (p = 0.04), but not of TNFalpha. CONCLUSION: Increased NO production by PBL in patients with MS is independent of the disease course. PMID- 14745061 TI - Epilepsy syndrome, focus location, and treatment choice affect testicular function in men with epilepsy. AB - OBJECTIVE: To evaluate testicular function in men with epilepsy treated with different antiepileptic drugs (AED) vs no drugs. METHODS: Two hundred men with epilepsy (mean age 36 years) were investigated. Inclusion criteria included one or no AED, no co-medication, and no endocrine co-morbidity. Findings were compared with those from 105 healthy men (mean age 33.9 years). Luteinizing hormone (LH), follicle-stimulating hormone (FSH), testosterone (T), free testosterone (free T), and inhibin B were measured. RESULTS: One hundred seventy eight men had focal epilepsy (117 of these had temporal lobe epilepsy [TLE]) and 22 idiopathic generalized epilepsy (IGE). Thirty-three men were not on AED; 124 men received enzyme-inducing AED. Free T was lowered in all patient subgroups, and the T/LH ratio was lowered in all groups except patients with IGE and patients receiving valproate (VPA). T was lower in patients with temporal than extratemporal focal epilepsy. Compared with TLE patients on carbamazepine (CBZ), patients treated with VPA had higher total T and lower LH values, resulting in a significantly lower T/LH ratio with CBZ. CONCLUSIONS: Epilepsy, especially TLE, adversely affects testicular endocrine function. CBZ may increase the negative effects of epilepsy on serum levels of reproductive hormones. PMID- 14745062 TI - Effect of epilepsy and antiepileptic drugs on male reproductive health. AB - BACKGROUND: Men with epilepsy have reduced fertility, and antiepileptic drugs may affect semen quality. Moreover, animal studies suggest that valproate (VPA) may be associated with testicular atrophy. OBJECTIVE: To evaluate reproductive function in men with epilepsy. METHODS: Sixty men with epilepsy and 41 control men were evaluated for their reproductive health. Fifteen men were taking carbamazepine (CBZ) and 18 men oxcarbazepine (OXC) for partial epilepsy, and 27 men were taking VPA for generalized epilepsy. Reproductive hormones were assayed from serum samples, semen analysis and ultrasonography of the testicles were performed, and testicular volume was calculated. RESULTS: Men on CBZ had low serum dehydroepiandrosterone sulfate concentrations (p < 0.001), and men on VPA had high concentrations of serum androstenedione (p < 0.001). The frequency of morphologically abnormal sperm was higher among CBZ-treated (p < 0.01), OXC treated (p < 0.05), and VPA-treated men (p < 0.01) than among the control men. Moreover, both CBZ and VPA were associated with poor motility of sperm (p < 0.05). In addition, the frequency of abnormally low sperm concentration was high in men on CBZ (p < 0.001), and the frequency of any sperm abnormality was high in men on VPA (p < 0.01). The VPA-treated men with abnormal sperm had smaller testicular volumes than the control men (p = 0.003). CONCLUSIONS: CBZ, OXC, and VPA are associated with sperm abnormalities in men with epilepsy. In addition, VPA-treated men with generalized epilepsy who have abnormal sperm may have reduced testicular volume. PMID- 14745063 TI - Risk of seizures while awake in pure sleep epilepsies: a prospective study. AB - OBJECTIVE: To estimate the risk of seizures while awake in pure sleep epilepsies in a prospective study. METHODS: From October 1, 1992, to October 31, 1996, all patients with pure sleep epilepsy presenting at a participating center were enrolled. Children with benign rolandic epilepsy and patients with frontal lobe epilepsy were excluded. Patients were followed for at least 2 up to 6 years. The primary endpoint was the occurrence of a seizure while awake. RESULTS: The authors enrolled 161 patients (64% male). Age at the time of inclusion ranged from 11 to 83 years (mean 43.2, median 39). Eighty-five percent presented generalized tonic clonic seizures. Both sleep seizures and seizures while awake were absent for 2 years after inclusion in the study in 78% of patients. Eighteen patients presented a seizure while awake. The estimated risk of a seizure while awake during 6 years of follow-up was 13% (95% CI 7 to 18%). Multivariate analysis showed that episodes of sudden withdrawal of therapy and a higher frequency of seizures at inclusion were associated with an increased risk of seizures while awake. The estimated risk of a seizure while awake in patients with none of the above risk factors was 6.5% (95% CI 1.5 to 11.3%) during 6 years of follow-up. CONCLUSION: The clinical picture of pure sleep epilepsies is characterized by a preponderance of generalized tonic clonic seizures, low seizure frequency, and a good prognosis. The risk of occurrence of a seizure while awake is low, particularly among patients with rare seizures and good compliance with the therapy. PMID- 14745064 TI - Depression but not seizure frequency predicts quality of life in treatment resistant epilepsy. AB - BACKGROUND: The two-thirds of patients with epilepsy who become seizure-free have a quality of life (QOL) similar to the general population. The major treatment challenge is patients with refractory epilepsy. Whereas neurologists typically focus on seizure reduction in the treatment of these patients, results of studies relating seizure frequency to QOL are conflicting. As depression is associated with reduced QOL in epilepsy and antiepileptic medications (AEDs) can cause depression, it is important to determine the relative roles of depression and seizure frequency in QOL in refractory epilepsy. METHODS: Prospective evaluation was conducted of patients with refractory epilepsy being admitted to an inpatient video-EEG monitoring unit. The impact of clinical variables (age, sex, marital status, seizure frequency, duration and type of seizure disorder, seizure localization, number of AEDs, depression) on QOL was analyzed. RESULTS: Depression was a powerful predictor of QOL (n = 122, beta = -35.8, p < 0.0001). No other variable predicted QOL. Depression was common (54%), severe (19% with suicidal thoughts), underdiagnosed (37%), and largely untreated (17% on antidepressants). CONCLUSIONS: Treatment of depression may be inadequately prioritized in the management of intractable epilepsy. PMID- 14745065 TI - Clinical and genetic studies in hereditary spastic paraplegia with thin corpus callosum. AB - BACKGROUND: A complicated form of recessive hereditary spastic paraplegias (HSPs) with thin corpus callosum (TCC) was first described in Japan, and most of the Japanese families showed linkage to chromosome 15q13-15. A recessive HSP locus (SPG11) has also been mapped to chromosome 15q13-15 in Italian and North American families with and without TCC, and it overlaps the region identified in the Japanese families. OBJECTIVE: To study clinically and genetically 12 Italian families with HSP and TCC. METHODS: The authors investigated 18 affected and 30 healthy individuals from 12 unrelated Italian families with recessive HSP-TCC. Clinical, neurophysiologic, and neuroradiologic studies were undertaken. All patients were negative for SPG7 mutations. Genetic linkage analyses were carried out with polymorphic DNA markers on 15q13-15. RESULTS: Five families were consistent with linkage, thus defining a 19.8-cM region between markers D15S1007 and D15S978, encompassing the SPG11 interval. In one consanguineous family, linkage could be firmly excluded, confirming genetic heterogeneity. Two families appeared not linked to the region, but this could not be firmly proved because of the small family size. The remaining four families were uninformative for linkage purposes. CONCLUSION: HSP-TCC is common in Italy. The phenotype is fairly homogeneous and is associated with impaired cognition. There are at least two loci for HSP-TCC, one of which is on chromosome 15q13-15. PMID- 14745066 TI - Germline mutational dynamics in myotonic dystrophy type 1 males: allele length and age effects. AB - BACKGROUND: The CTG repeat expansion causing myotonic dystrophy type 1 is unstable in the germline, and frequent intergenerational length changes are observed, giving rise to the unusual genetics of the disorder. The repeat is also somatically unstable, and expanded alleles accumulate throughout life, thus compromising simple measures of intergenerational stability. OBJECTIVE: To gain a better understanding of the intergenerational dynamics of the DM1 repeat in the male germline. METHODS: We used sensitive small pool PCR procedures to analyze sperm and somatic DNA from 22 DM1 men of different ages, CTG repeat length, and clinical form. RESULTS: High levels of repeat length variation heavily biased toward further expansions were observed in the sperm of all DM1 men. Progenitor allele length was revealed as a major modifier of interindividual variation, with the largest length changes observed for premutation and protomutation alleles and the highest frequency of contractions in full mutation alleles. However, despite clear increases in the degree of somatic mosaicism, no differences were observed in replicate sperm samples obtained from two men during a 4-year period. CONCLUSIONS: Progenitor allele length is a major modifier of the mutational dynamics of the DM1 repeat in the male germline, but surprisingly age is not. Therefore, other as yet unidentified modifiers must be responsible for the considerable residual interindividual variation that cannot be accounted for by these factors. PMID- 14745067 TI - Dietary intake of fatty acids and fish in relation to cognitive performance at middle age. AB - OBJECTIVE: To examine the associations of fatty acid and fish intake with cognitive function. METHODS: Data are from a cross-sectional population-based study among 1,613 subjects ranging from 45 to 70 years old. From 1995 until 2000, an extensive cognitive battery was administered and compound scores were constructed for memory, psychomotor speed, cognitive flexibility (i.e., higher order information processing), and overall cognition. A self-administered food frequency questionnaire was used to assess habitual food consumption. The risk of impaired cognitive function (lowest 10% of the compound score) according to the energy adjusted intake of fatty acids was assessed with logistic regression, adjusting for age, sex, education, smoking, alcohol consumption, and energy intake. RESULTS: Marine omega-3 polyunsaturated fatty acids (PUFA) (eicosapentaenoic acid and docosahexaenoic acid) were inversely related to the risk of impaired overall cognitive function and speed (per SD increase: OR = 0.81, 95% CI 0.66 to 1.00 and OR = 0.72, 95% CI 0.57 to 0.90). Results for fatty fish consumption were similarly inverse. Higher dietary cholesterol intake was significantly associated with an increased risk of impaired memory and flexibility (per SD increase: OR = 1.27, 95% CI 1.02 to 1.57 and OR = 1.26, 95% CI 1.01 to 1.57). Per SD increase in saturated fat intake, the risk of impaired memory, speed, and flexibility was also increased, although not significantly. CONCLUSIONS: Fatty fish and marine omega-3 PUFA consumption was associated with a reduced risk and intake of cholesterol and saturated fat with an increased risk of impaired cognitive function in this middle-aged population. PMID- 14745068 TI - Spontaneous carotid artery dissection. PMID- 14745069 TI - Clinical and MRI outcome after autologous hematopoietic stem cell transplantation in MS. AB - The authors report the outcome of 14 patients with severe multiple sclerosis treated with autologous hematopoietic stem cell transplantation (AHSCT) after a median follow-up period of 3 years. The 3-year actuarial probability of progression-free survival was 85.7% and that of disease activity-free survival was 46.4%. On MRI, no T1-enhanced lesions were detected after AHSCT. The mean change in T2 lesion volume from baseline to the third year was -20.2% and that of the corpus callosum area was -12.7%; 50% of this reduction was seen during the first year. PMID- 14745070 TI - Valproic acid has no effect on pain in polyneuropathy: a randomized, controlled trial. AB - The aim of this randomized, double-blind, placebo-controlled cross-over study was to test whether valproic acid relieves painful polyneuropathy. Thirty-one patients completed two treatment phases of 4 weeks' duration with valproic acid (1,500 mg daily) and placebo. There was no significant difference between valproic acid and placebo in the rating of total pain (median, 5 in the valproic acid period vs 6 in the placebo period; p = 0.24) or in individual pain ratings. PMID- 14745071 TI - Lateralizing value of Todd's palsy in patients with epilepsy. AB - The authors retrospectively investigated the value of Todd's palsy (TP) in lateralizing the hemisphere of seizure onset in patients admitted for video-EEG monitoring in a tertiary epilepsy center. In 29 patients, a postictal hemiparesis was observed. TP always occurred contralateral to the epileptogenic hemisphere in 27 patients (93%). In the remaining two patients, the seizure onset could not be lateralized. In some patients, TP occurred after a seizure without focal motor features or secondary generalization. PMID- 14745072 TI - The impact of pregnancy and childbirth on the metabolism of lamotrigine. AB - This study was performed to clarify alterations in lamotrigine (LTG) clearance during pregnancy and childbirth. Fourteen women on LTG monotherapy had LTG concentration samples obtained before conception and monthly. LTG apparent clearance, weight-adjusted relative clearance, and percentages of baseline clearance significantly differed between preconception baseline and each trimester and between trimesters. LTG clearance progressively increased until 32 weeks' gestational age, reaching a peak of >330% of baseline, and then began to decline. PMID- 14745073 TI - Zonisamide treatment for symptomatic infantile spasms. AB - The authors evaluated zonisamide for symptomatic infantile spasms in 23 patients. Spasm cessation, EEG evolution, and tolerability were assessed for a mean duration of 6.5 months. Six patients (26%) had complete control with cessation of spasms and clearing of hypsarrhythmia. Mean latency time from onset of zonisamide treatment to complete spasm control was 19 days. There were no discontinuations due to adverse effects. PMID- 14745074 TI - Reversible language regression as an adverse effect of topiramate treatment in children. AB - Profound language regression developed in three children with epilepsy 4 to 28 weeks after beginning topiramate (TPM). TPM was administered as an adjunctive antiepileptic drug at doses of 2.5 to 6.0 mg/kg/day. Language functions recovered while TPM was being reduced in dose or stopped. PMID- 14745075 TI - Low androgenization index in elderly women and elderly men with Alzheimer's disease. AB - The authors investigated whether testosterone levels and testosterone availability differ between older lean subjects with and without Alzheimer's disease (AD). Sex hormone binding globulin (SHBG) and estradiol levels were higher, whereas the free androgenization index (FAI) was lower, in lean subjects with AD than in lean subjects without AD. Factors involved in the increase of SHBG secretion could have an important role in the lower testosterone availability of subjects with AD. PMID- 14745076 TI - PEN2 is not a genetic risk factor for Alzheimer's disease in a large family sample. AB - PEN2 is a reasonable Alzheimer's disease (AD) candidate gene because it is a necessary component of the gamma-secretase complex that generates beta-amyloid peptide. Moreover, its gene (PEN2) maps to a highly significant linkage region on chromosome 19q13. Four common polymorphisms in PEN2 were tested for genetic association with AD in a large and carefully ascertained AD family sample (789 subjects from 202 nuclear families) using single-locus and haplotype-based analyses. These results do not suggest PEN2 to be a major AD risk factor. PMID- 14745077 TI - Association between tumor necrosis factor-alpha promoter polymorphism and Alzheimer's disease. AB - Tumor necrosis factor-alpha (TNFalpha) gene polymorphisms have been reported to be associated with Alzheimer's disease (AD) in Caucasian populations. Three TNFalpha polymorphisms (-857, -863, and -1,031) were studied in a Chinese population. A high-risk TNFalpha haplotype (-1,031C-863C-857C) with an odds ratio of 2.54 (95% CI 1.37 to 4.79) for AD was identified. No interaction effect of APOE and TNFalpha genotypes was found, but both acted as important risk factors for AD. PMID- 14745078 TI - White matter lesions and hippocampal atrophy in Alzheimer's disease. AB - White matter lesions (WML) and hippocampal atrophy (HA) on MRI commonly co-occur in Alzheimer's disease (AD) and are thought to play a role in the etiology of AD. It is still not known whether WML and HA are independent or related. The authors investigated the relation between WML and HA in 179 patients with probable AD who had a cerebral MRI. A linear relation was found between WML and HA, especially for WML in the frontal and parieto-occipital regions. The results suggest that vascular pathology and typical AD pathology (HA) are related. PMID- 14745079 TI - Polymorphisms within the prion (PrP) and prion-like protein (Doppel) genes in AD. AB - The authors present a study on the association of PRNP and PRND gene polymorphisms with the occurrence and age at onset of Alzheimer's disease (AD). DNA from 79 Polish patients with probable AD and 107 healthy control subjects was studied. The PRNP codon 129 homozygosity seemed to be associated with the occurrence of AD: In AD patients, the percentage of Val/Val and Met/Met genotypes was higher than in the control subjects. A significant difference appeared also between early-onset (<70 years) and late-onset (> or = 70 years) AD patients in the PRND genotypes. PMID- 14745080 TI - POLG mutations causing ophthalmoplegia, sensorimotor polyneuropathy, ataxia, and deafness. AB - The authors identified two novel heterozygous missense transitions in the gene for the mitochondrial polymerase gammaA subunit (POLG) in a family with an autosomal recessive syndrome comprising progressive external ophthalmoplegia (PEO), polyneuropathy, ataxia, sensorineural hearing loss, and affective disorders. These mutations were not detected in 120 healthy control subjects. PMID- 14745081 TI - Widespread increased expression of the DNA repair enzyme PARP in brain in ALS. AB - Expression of the DNA repair enzyme poly(ADP-ribose) polymerase (PARP) is a known response to oxidative damage of DNA. In ALS brain, PARP expression by western analyses was increased in the motor cortex, parietal cortex, and cerebellum. PARP immunostaining in the motor cortex was increased in ALS neurons and subcortical glia and macrophages. Importantly, there was widespread increased PARP expression in neurons in the parietal cortex and cerebellum, regions that are typically clinically unaffected in ALS, suggesting widespread oxidative stress. PMID- 14745082 TI - Multiparametric MRI in a patient with adult-onset leukoencephalopathy with vanishing white matter. AB - This is a multiparametric MR study of the first reported patient with adult-onset genetically confirmed vanishing white matter (VWM) disease. It shows that, despite the presence of a severe and diffuse damage of the brain WM, brain gray matter, and cervical cord tissue, the cortical adaptive capacities were relatively preserved. Interpatient variability of brain plasticity may contribute to the known phenotypic variation of patients with VWM disease. PMID- 14745083 TI - Spinocerebellar ataxia type 5: clinical and molecular genetic features of a German kindred. AB - The authors report a German family with autosomal dominant cerebellar ataxia tightly linked to the spinocerebellar ataxia type 5 (SCA5) locus (multipoint lod score 5.76). The phenotype is characterized by a purely cerebellar syndrome with a downbeat nystagmus occurring prior to the development of other features. Imaging studies demonstrated cortical cerebellar atrophy. Progression is slow even in patients with a disease onset during the second decade. The age at onset varies from 15 to 50 years. PMID- 14745084 TI - The safety and tolerability of a mixed lineage kinase inhibitor (CEP-1347) in PD. AB - CEP-1347 is an inhibitor of members of the mixed lineage kinase family, key signals triggering apoptotic neuronal death. The authors performed a randomized, blinded, placebo-controlled study assessing the safety, tolerability, pharmacokinetics, and acute symptomatic effects of CEP-1347 in 30 patients with Parkinson's disease (PD). In this short-term study, CEP-1347 was safe and well tolerated. It had no acute effect on parkinsonian symptoms or levodopa pharmacokinetics, making it well suited for larger and longer studies of its potential to modify the course of PD. PMID- 14745085 TI - Tic douloureux or "tic dentaire". PMID- 14745086 TI - Tamoxifen: a new risk factor for cerebral sinus thrombosis. PMID- 14745087 TI - Reversible abnormalities of DWI in high-altitude cerebral edema. PMID- 14745088 TI - Somatic mosaicism in Charcot-Marie-Tooth type X disease. PMID- 14745089 TI - Therapeutic effect and [123I]IMP SPECT findings of sodium dichloroacetate in a patient with MELAS. PMID- 14745090 TI - Levetiracetam improves paroxysmal symptoms in a patient with stiff-person syndrome. PMID- 14745091 TI - Personal History: the pelican, in brief. PMID- 14745092 TI - Effect of antiepileptic drugs on bone density in ambulatory patients. PMID- 14745093 TI - The prevalence of migraine in neurologists. PMID- 14745094 TI - Slowing Parkinson's disease progression: recent dopamine agonist trials. PMID- 14745095 TI - Pure dysarthria due to small cortical stroke. PMID- 14745096 TI - Patient page. Surgery for patients with Parkinson disease. PMID- 14745097 TI - Effects of nitric oxide on proprioceptive signaling. AB - We have analysed the effects of the neuromodulator nitric oxide (NO) on proprioceptive information processing by ascending intersegmental interneurons that form part of the local circuits within the terminal abdominal ganglion of the crayfish. NO modulates the synaptic inputs to ascending interneurons, enhancing the amplitude of class I interneurons and reducing the amplitude of class II interneurons. Repetitive proprioceptive stimulation leads to rapid depression in a specific set of identified interneurons but not in others. Bath application of a nitric oxide scavenger, PTIO, causes a significant decrease in the rate of depression of the interneurons showing a rapid depression, independent of interneuron class, but has no effect on the dynamic responses of the interneurons that show little initial depression. These results indicate that NO exerts multiple effects at the very first stage of synaptic integration in local circuits. PMID- 14745098 TI - Functional changes of cricket giant interneurons caused by chronic unilateral cercal ablation during postembryonic development. AB - One of a pair of cerci was ablated in the first-, fourth- and last-instar nymphs of the cricket, Gryllus bimaculatus. The insects were then reared until the final molt, after which the intensity-response (I-R) relationships for four giant interneurons (GIs) 8-1, 9-1, 9-2 and 9-3 with regard to a controlled air current stimulus were measured. In order to examine the functional changes during postembryonic development and the differences in the physiological plasticity of GIs between nymphs and adults, the obtained I-R curves for each GI were compared with those measured in intact and unilaterally cercus-ablated adult crickets. Each GI showed a distinctive change in response magnitudes after the long-term unilateral cercal ablation. In most cases, the I-R curves for each GI in the crickets ablated from nymphal periods were different from those in the adult crickets mentioned above. Moreover, the pattern of change in response magnitude was different from GI to GI. In contrast to these observations, it was reported that some important characteristics of the wind-evoked escape behavior such as relative occurrence and escape direction in unilaterally cercus-ablated crickets investigated after a long-term rearing were almost identical with those in intact crickets. Therefore, the results obtained in the present study suggest that functional changes occur not only in GIs but also in many other neural elements in the escape-eliciting system in order to maintain the features of wind-evoked escape behavior. PMID- 14745099 TI - Dual effects of dopamine on the adult heart of the isopod crustacean Ligia exotica. AB - In the adult heart of the isopod crustacean Ligia exotica, the cardiac ganglion acts as the primary pacemaker with the myocardium having a latent pacemaker property. We show several lines of evidence that dopamine modulates the heartbeat of adult L. exotica affecting both pacemaker sites in the heart. Dopamine caused positive chronotropic (frequency increase) and inotropic (amplitude increase) effects on the heartbeat in a concentration dependent manner. The time courses of these effects were considerably different and the inotropic effect appeared later and lasted longer than the chronotropic effect. Dopamine rapidly increased the frequency of the bursting activity in the cardiac ganglion neurons and each impulse burst of the cardiac ganglion was always followed by a heartbeat. Moreover, dopamine slowly increased the amplitude and duration of the action potential plateau (plateau potential) of the myocardium. When the myocardial pacemaker activity was induced by application of tetrodotoxin, which suppresses cardiac ganglion activity, dopamine slowly increased the amplitude and duration of the myocardial plateau potential while decreasing its frequency. These results suggest that dopamine modulates the heartbeat in adult L. exotica producing a dual effect on the two pacemaker sites in the heart, the cardiac ganglion and myocardium. PMID- 14745100 TI - The changes of gene expression in honeybee (Apis mellifera) brains associated with ages. AB - Honeybee (Apis mellifera) worker bees (workers) are known to perform wide variety of tasks depending on their ages. The worker's brains also show the activity and behavior-dependent chemical and structural plasticity. To test if there are any changes of gene expression associated with different ages in the worker brains, we compared the gene expression patterns between the brains of newly emerged bees and old foraging workers (foragers) by macroarray analysis. The expression of genes encoding signal transduction pathway components, ion channels, and neurotransmitter transporters is elevated in the old forager brains, suggesting that the neuronal activities would be enhanced. The mRNA levels of cell adhesion protein, transcription related factors, and plasma membrane associated proteins are also increased in the old forager brains. Meanwhile, the mRNA level of one putative cell adhesion protein is decreased in the old forager brains. These results thus suggest that the dramatic changes of gene expression occur in honeybee brains associated with ages. PMID- 14745101 TI - Entrainment of the circatidal swimming activity rhythm in the cumacean Dimorphostylis asiatica (Crustacea) to 12.5-hour hydrostatic pressure cycles. AB - The cumacean Dimorphostylis asiatica (Crustacea) exhibits a circatidal swimming activity rhythm. The animals were exposed to a 12.5 hr sinusoidal change of hydrostatic pressure of 0.3 atm amplitude in the laboratory. Under constant dark conditions, most of the specimens were entrained to a daily bimodal swimming activity rhythm by the hydrostatic pressure cycle. A small number of individuals exhibited a unimodal daily rhythm, with no apparent entraining from the administered cycles. A marked feature was a flexible phase relationship between the entrained daily bimodal rhythm and the hydrostatic pressure cycles: the swimming activity of most of the specimens occurred around the pressure decreasing phase, but for a small number of individuals it coincided with the pressure-increasing phase. Such flexibility suggests a weak entraining effect of hydrostatic pressure on the circatidal rhythm of this species. When exposed to 24 hr light-dark cycles and a hydrostatic pressure cycle simultaneously, the specimens exhibited a rhythmic activity entrained by the hydrostatic pressure cycle during the dark period, which closely resembles the temporal activity pattern of this species in the field. The light cycles entrained the swimming activity via direct inhibition and induction of activity (i.e., masking). Under light-dark conditions, the specimens exhibited activity on the pressure increasing phase more frequently compared with specimens kept in constant darkness. PMID- 14745102 TI - Notes on a discrepancy in mitochondrial DNA and allozyme differentiation in a pond frog Rana nigromaculata. AB - Analyses of complete 1143-base pair sequence of the mitochondrial cytochrome b gene demonstrated a sister relationship between Japanese R. nigromaculata and Korean R. plancyichosenica, but not with Korean R. nigromaculata, while the allozyme data strongly supported the monophyly of the Korean and Japanese populations of R. nigromaculata. We surmise this discordance to be the result of the inheritance of introduced mtDNA and the dilution of introduced nuclear DNA in mixed lineages after past hybridization and genome introgression between the two species, although the direction of introgression is unknown. PMID- 14745103 TI - Inter-male mating-like behavior in the domesticated house musk shrew, Suncus murinus. AB - In the present study, inter-male interaction of the domesticated house musk shrew was observed in detail under laboratory conditions. In most cases, during inter male interaction, male house musk shrews exhibited a sequence of behavior items including tail-wagging, following, mounting and thrusting. In the minority of cases, males did not progress beyond following. Offensive behavior was not sufficiently violent to cause injury. It appeared that role assignment was decided by contact manner and vocalization. One of fundamental characters of this animal made a start of following, in which one shrew followed another, who touched and then separated. Role assignment (i.e., which male led and which followed) was decided in status battle. Roles were often reversed during following. Following behavior was maintained by 'polite' mutual contact, and the interaction progressed to thrusting in the majority cases. After role assignment, the variation in contact manner decreased. The ratio of time spent in front-and behind contact to that spent in multi-lateral contact increased when both males commenced following behavior simultaneously. This ratio was maintained until the following male snapped after he finished thrusting. Even if the following male did not reach thrusting, he mounted the preceding male. The pairs who did not reach thrusting repeated following behavior or mounting. In those cases, while one male concentrated on touching the other to maintain following, the other attempted to divert attention from the following behavior. Male shrews were able to reach thrusting irrespective of sex. PMID- 14745104 TI - Identification of honeybee antennal proteins/genes expressed in a sex- and/or caste selective manner. AB - We identified three candidate proteins/genes involved in caste and/or sex specific olfactory processing in the honeybee Apis mellifera L., that are differentially expressed between the antennae of the worker, queen, and drone honeybees using SDS-polyacrylamide gel electrophoresis or the differential display method. A protein was identified, termed D-AP1, that was expressed preferentially in drone antennae when compared to those of workers. cDNA cloning revealed that D-AP1 is homologous to carboxylesterases. Enzymatic carboxylesterase activity in the drone antennae was higher than in the workers, suggesting its dominant function in the drone antennae. In contrast, two proteins encoded by genes termed W-AP1 and Amwat were expressed preferentially in worker antennae when compared to those of queens. W-AP1 is homologous to insect chemosensory protein, and Amwat encodes a novel secretory protein. W-AP1 is expressed selectively in worker antennae, while Amwat is expressed both in the antennae and legs of the workers. These findings suggest that these proteins are involved in the antennal function characteristic to drone or worker honeybees. PMID- 14745105 TI - Crystalline calcium phosphate and magnetic mineral content of Daphnia resting eggs. AB - Daphnia is a key crustacean zooplankton of freshwater food chains. One factor that ensures successful propagation is the Daphnia resting eggs, which are able to retain structural integrity under extreme conditions. Until recently little was known about the chemical composition, microanatomy, and physical properties of the egg itself. The current study demonstrates that the resting eggs: (1) have shells that are made up of crystalline calcium phosphate and include a honeycombed structure, and (2) contain magnetic material having properties consistent with magnetite. These properties of the resting eggs may ensure Daphnia survival in harsh environments. PMID- 14745106 TI - Development and neural organization of the tornaria larva of the Hawaiian hemichordate, Ptychodera flava. AB - We report scanning and transmission electron microscopic studies of the early development of the Hawaiian acorn worm, Ptychodera flava. In addition, we provide an immunohistochemical identification of the larval nervous system. Development occurs and is constrained within the stout chorion and fertilization envelope that forms upon the release of the cortical granules in the cytoplasm of the egg. The blastula consists of tall columnar blastomeres encircling a small blastocoel. Typical gastrulation occurs and a definitive tornaria is formed compressed within the fertilization envelope. The young tornaria hatches at 44 hr and begins to expand. The major circumoral ciliary band that crosses the dorsal surface and passes preorally and postorally is well developed. In addition, we find a nascent telotroch, as well as a midventral ciliary band that is already clearly developed. The epithelium of tornaria is a mosaic of monociliated and multiciliated cells. Immunohistochemistry with a novel neural marker, monoclonal antibody 1E11, first detects nerve cells at the gastrula stage. In tornaria, 1E11 staining nerve cells occur throughout the length of the ciliary bands, in the apical organ, in a circle around the mouth, in the esophageal epithelium and in circumpylorus regions. Axon(s) and apical processes extend from the nerve cell bodies and run in tracks along the ciliary bands. Axons extending from the preoral and postoral bands extend into the oral field and form a network. The tornaria nervous system with ciliary bands and an apical organ is rather similar to the echinoderm bipinnaria larvae. PMID- 14745107 TI - Disturbance of plasma melatonin profile by high dose melatonin administration inhibits testicular maturation of precocious male masu salmon. AB - We have previously shown that the testicular development of underyearling male masu salmon Oncorhynchus masou reared under a long photoperiod was accelerated by oral melatonin treatment (0.5 mg melatonin/kg body weight/day), suggesting that melatonin mediates photoperiodic signaling. In this study, we further examined the effects of a disturbance in the plasma melatonin profile on gonadal development in underyearling male masu salmon by administering a higher dose of melatonin. Fish randomly selected in June were divided into two groups. They were reared under a light:dark (LD) cycle of 16:8 (lights on 04:00-20:00 hr) and fed with pellets sprayed with melatonin or vehicle twice a day at 08:30 and at 15:30 hr (7.5 mg melatonin/kg body weight/day) until October. Fish were sampled on Day 0, 25, 60, 90 and 120. The plasma melatonin levels were high in the dark phase and low in the light phase in the control group, while they were constantly high with no significant change in the melatonin-treated group. Melatonin treatment had inhibitory effects on the gonadosomatic index and plasma testosterone levels. Pituitary salmon gonadotropin-releasing hormone content and luteinizing hormone content were significantly lower in the melatonin-treated group on Day 60 and 90, respectively. These results indicate that the plasma melatonin profile is important for mediating photoperiodic signals that regulate brain-pituitary gonadal axis in underyearling precocious male masu salmon. PMID- 14745108 TI - Glucose-stimulated somatostatin gene expression in the Brockmann bodies of rainbow trout (Oncorhynchus mykiss) results from increased mRNA transcription and not from altered mRNA stability. AB - Previously, we showed that glucose increases the steady-state levels of the mRNAs encoding two distinct preprosomatostatins (each containing [Tyr7, Gly10] somatostatin-14 at their C-termini; denoted PPSS II' and PPSS II") in the endocrine pancreas (Brockmann body) of rainbow trout (Oncorhynchus mykiss). In the present study, isolated islet cells were used to determine whether glucose stimulated expression resulted from altered rates of transcription and/or from changes in RNA stability. Nuclear run-on assays indicated that the number of PPSS II nascent transcripts were significantly higher in nuclei isolated from islet cells cultured in 10 mM glucose compared to those isolated from cells incubated in 4 mM glucose. High glucose (10 mM) did not, however, affect the stability of PPSS II mRNAs. These results indicate that glucose-stimulated somatostatin expression in the Brockmann bodies of rainbow trout results from increased endogenous mRNA transcription and not from altered mRNA stability. PMID- 14745109 TI - A new hermit crab species of Pylopaguropsis (Crustacea: Decapoda: Anomura: Paguridae) from the western Pacific, and supplemental note on P. laevispinosa McLaughlin and Haig. AB - Pylopaguropsis vicina, a new species of the Paguridae, is described and illustrated from southern Japan and Indonesia. This new species is most similar to P. laevispinosa McLaughlin and Haig, but is distinguished by the armature of the chelae and ambulatory dactyli, spination of the telson, and stripe pattern of the pereopods. Reexamination of specimens referred to P. laevispinosa by recent authors has shown these assignments to be incorrect. A brief supplemental description is provided for P. laevispinosa in order to clarify its systematic status. An amended key to species of the genus Pylopaguropsis is presented. PMID- 14745110 TI - A new species Megabruchidius sophorae (Coleoptera, Bruchidae), feeding on seeds of Styphnolobium (Fabaceae) new to Bruchidae. AB - A new species Megabruchidiussophorae (Insecta, Coleoptera) is described from Japan (Honshu). The larval host of this bruchid is the seeds of the tree legume 'enju', or chinese scholar tree, Styphnolobium japonicum (a senior synonym of Sophora japonica), which is a new host genus to Bruchidae. Styphnolobium is positioned basally in molecular phylogeny of the leguminous subfamily Papilionoideae. Other members of Megabruchidius are known to feed on Gleditsia, the tree legumes that belong to the most ancestral subfamily Caesalpinioideae. Therefore, Megabruchidius utilizes ancestral groups of legumes as its host plants. Megabruchidius has been inferred to be ancestral, based on its behavior. The character state of the host for this third Megabruchidius species supports that the genus is ancestral, at least in the subfamily Bruchinae. We also reviewed the genera closely related to Megabruchidius, i.e., Bruchidius and Sulcobruchus in Bruchidini, and wrote a key to the species in the genus Megabruchidius. PMID- 14745111 TI - Molecular phylogeny of arctoids (Mammalia: Carnivora) with emphasis on phylogenetic and taxonomic positions of the ferret-badgers and skunks. AB - Phylogenetic relationships among the ferret-badger Melogale moschata, the skunk Mephitis mephitis, and 21 other arctoid carnivorans, representing Mustelidae (Mustelinae: Mustela, Martes, Gulo; Lutrinae: Enhydra; Melinae: Meles), Procyonidae (Procyon), and Ursidae (Ursus, Melursus), were evaluated through maximum-parsimony phylogenetic analysis of concatenated partial nucleotide sequences of the nuclear recombination-activating gene 1 (RAG1) and gene encoding interphotoreceptor retinoid-binding protein (IRBP). The analysis strongly supports Melogale as more closely related to a musteline-lutrine clade (containing Mustela and Enhydra) than to Meles or another musteline clade containing Martes and Gulo (causing Melinae and Mustelinae, as traditionally circumscribed, to be nonmonophyletic). This, together with known morphological and karyological evidence for nonmeline affinities of Melogale, justify the exclusion of the ferret-badgers from the monophyletic Melinae. Therefore, we recommend that Melogale be classified in a distinct mustelid subfamily, the monotypic Helictidinae. Our analysis also strongly supports an outgroup position of the skunks to a clade containing Procyonidae and the nonmephitine Mustelidae (causing Mustelidae, as traditionally circumscribed, to be paraphyletic). This position of the skunks agrees with results of most previous genetic studies. However, it is contradicted by known morphological evidence from both living and fossil taxa, as well as genetic evidence from protein electrophoresis. These consistently support the traditional placement of the skunks within the monophyletic Mustelidae (recently in a close relationship to Lutrinae). Therefore, we consider the recent elevation of the skunks to the level of family as premature, and recommend that this clade be left at the subfamily level (Mephitinae) within the family Mustelidae, pending further evidence. PMID- 14745112 TI - Three-finger alpha-neurotoxins and the nicotinic acetylcholine receptor, forty years on. AB - The discovery, about forty years ago, of alpha-bungarotoxin, a three-finger alpha neurotoxin from Bungarus multicinctus venom, enabled the isolation of the nicotinic acetylcholine receptor (nAChR), making it one of the most thoroughly characterized receptors today. Since then, the sites of interaction between alpha neurotoxins and nAChRs have largely been delineated, revealing the remarkable plasticity of the three-finger toxin fold that has optimally evolved to utilize different combinations of functional groups to generate a panoply of target specificities to discern subtle differences between nAChR subtypes. New facets in toxinology have now broadened the scope for the use of alpha-neurotoxins in scientific discovery. For instance, the development of short, combinatorial library-derived, synthetic peptides that bind with sub-nanomolar affinity to alpha-bungarotoxin and prevent its interaction with muscle nAChRs has led to the in vivo neutralization of experimental alpha-bungarotoxin envenomation, while the successful introduction of pharmatopes bearing "alpha-bungarotoxin-sensitive sites" into toxin-insensitive nAChRs has permitted the use of various alpha neurotoxin tags to localize and characterize new receptor subtypes. More ambitious strategies can now be envisaged for engineering rationally designed novel activities on three-finger toxin scaffolds to generate lead peptides of therapeutic value that target the nicotinic pharmacopoeia. This review details the progress made towards achieving this goal. PMID- 14745113 TI - YC-1, a nitric oxide-independent activator of soluble guanylate cyclase, inhibits the spontaneous contractions of isolated pregnant rat myometrium. AB - The aim of this study was to investigate the effect of YC-1 (3-(5'-hydroxymethyl 2'-furyl)-1-benzyl indazole) on spontaneous contractions and levels of cyclic GMP (cGMP) of myometrial strips isolated from pregnant rats. It is a nitric oxide independent soluble guanylate cyclase activator. Myometrial strips were obtained from eight pregnant Wistar albino rats and were mounted in organ baths for the recording of isometric tensions. We evaluated the effect of increasing concentrations of YC-1 on spontaneous myometrial contractions and on contractions of myometrial smooth muscle pretreated with methylene blue (10(-5) M), tetraethylammonium chloride (TEA) (3 x 10(-4) M), and glibenclamide (10(-6) M). YC-1 (10(-9) - 3 x 10(-5) M) concentration-dependently decreased the amplitude and frequency of spontaneous contractions of myometrial strips. The inhibition of the amplitude and frequency of spontaneous contractions by YC-1 were antagonized with methylene blue (10(-5) M) and TEA (3 x 10(-4) M), but they were not changed by glibenclamide (10(-6) M); however, the antagonistic effect of methylene blue was significantly more than that of TEA (P<0.05). We also evaluated the effect of YC-1 on the levels of cGMP in myometrial strips obtained from pregnant rat uterine horns. YC-1-stimulated myometrial strips showed an excessive elevation in myometrial cGMP that declined slowly during the subsequent washout period. These results show that YC-1 decreases spontaneous contractile activity of myometrial strips isolated from pregnant rat and causes elevation of myometrial cGMP levels in vivo. This effect of YC-1 is significantly reduced by the methylene blue and TEA, suggesting the activation of soluble guanylate cyclase and Ca(2+)-sensitive K(+) channels as the mechanisms of action. PMID- 14745114 TI - A novel method to quantify calcium response pattern and oscillation using fura2 and acridine orange. AB - To study calcium imaging data of cell populations that have various response patterns in peak amplitude and frequency of calcium oscillation in response to stimulation, comprehensive characterization based on statistical analysis of each response is important. In cultures of cells that are flat and in contact with each other, it is difficult to distinguish individual cells in calcium imaging data. We have developed a novel method to determine areas corresponding to individual cells in calcium imaging data. Rat neonatal cerebral astrocytes were filled with the calcium indicator Fura2, stained with acridine orange, and illuminated with UV light. The cell nuclei were clearly visualized. In addition, the images of these nuclei were useful for analyzing concentration-dependent alteration of calcium oscillation of cultured astrocytes in response to glutamate. This novel method may be useful for studying factors affecting calcium response patterns of cultured cell populations, including culture conditions, stimulus paradigms, and synthetic compounds. PMID- 14745116 TI - Detrimental role of corticotropin-releasing factor on the decrease of CA1 field potential induced by in vitro ischemia in rat hippocampal slices. AB - This experiment was designed to test the hypothesis that endogenous corticotropin releasing factor (CRF) contributes to the neurodegenerative process following an ischemic insult. To test this hypothesis, the effects of chronic intracerebroventricular administration of CRF or astressin, a CRF-receptor antagonist, on the decrease in the Schaffer collateral-CA1 field potential induced by hypoxia/hypoglycemia (ischemia), were tested in rat hippocampal slices. The chronic treatment with CRF had a significant exacerbating effect on the 10-min ischemia, a condition that did not affect the evoked synaptic response in the hippocampal CA1 area, as compared to vehicle-treated rats. On the other hand, astressin had a significant ameliorative effect on the 15-min ischemia induced reduction of the evoked synaptic response in the hippocampal CA1 area. These findings suggest that CRF accelerates hippocampal ischemic vulnerability induced by hypoxia and hypoglycemia. PMID- 14745115 TI - Effect of nicotine-induced corticosterone elevation on nitric oxide production in the passive skin arthus reaction in rats. AB - To elucidate the anti-inflammatory action of nicotine-induced corticosterone elevation on the passive skin Arthus reaction (PSAR), we investigated the inflammatory process in the PSAR. The polymorphonuclear leukocyte (PMNs) infiltration was observed just before as well as after elicitation by measuring extractable myeloperoxidase. The plasma exudation was significantly inhibited by anti-rat tumor necrosis factor (TNF)-alpha antibody (5 microg/site, i.d.) at the time of sensitization or by superoxide dismutase (52500 units/kg, i.p.) 1 h before elicitation or N(G)-nitro-L-arginine-methyl ester (100 mg/kg, i.v.) just at elicitation. Pretreatment with a single injection of nicotine (0.8 mg/kg, i.p.) 30 min before elicitation suppressed the plasma exudation but not the PMNs infiltration. This nicotine-induced decreasing effect was abolished in animals supplemented with L-arginine (300 mg/kg, i.v.) just at elicitation. The production of nitric oxide (NO) in peritoneal PMNs derived from an animal injected peritoneally with oyster glycogen was significantly suppressed by pretreatment with nicotine (0.8 mg/kg, i.v.) 30 min prior to harvesting. This inhibitory action of nicotine was abolished in animals pretreated with mifepristone (30 mg/kg, s.c.), a glucocorticoid receptor antagonist. These findings indicate that a single systematic administration of nicotine may attenuate the plasma exudation in the PSAR by suppressing the production of NO in the PMNs primed with TNF-alpha via nicotine-induced endogenous glucocorticoid. PMID- 14745117 TI - The action mode of lysophosphatidylcholine in human monocytes. AB - To elucidate the action and signal transduction of lysophosphatidylcholine (LPC), we challenged a set of LPC on U937 human monocytes and found that LPC mobilized Ca(2+). The Ca(2+) response was not blocked by pertussis toxin, an inhibitor of G(i/o) proteins, or by U73122, a phospholipase C inhibitor. Furthermore, the response was totally blocked by addition of EGTA to the extracellular media, suggesting that Ca(2+) influx across the plasma membrane was the only source of LPC-induced Ca(2+) response in the U937 cells. 16:0 and 18:0 LPC induced similar responses. Recently it has been suggested that two G protein-coupled receptors function as LPC receptors in the plasma membrane. RT-PCR analysis indicated that neither the G2A receptor nor the GPR4 receptor is expressed in the U937 monocytes. Our data suggests that another action mechanism of LPC may be involved in the LPC response in the U937 cells. PMID- 14745118 TI - Estrogenic effects of Pueraria mirifica on the menstrual cycle and hormone related ovarian functions in cyclic female cynomolgus monkeys. AB - This study investigated the estrogenic effect of Pueraria mirifica (P. mirifica) on menstrual cycle length and hormone-related ovarian function. Nine normal cyclic monkeys (Macaca fascicularis) were separated into 3 groups; each group was force fed with a single dose of 10, 100, and 1,000 mg of P. mirifica. The experimental schedule was separated into the pre-treatment and post-treatment periods. Blood samples were collected on days 3, 9 - 14, 19, 24, 29, and every 10 days until the next menstruation for one and two menstrual cycles during two consecutive periods and assayed for serum levels of gonadotropins and ovarian hormones. The result showed a significant increase in lengths of the follicular phase and total menstrual cycle in monkeys treated with 1,000 mg of P. mirifica, but no change in menstrual cycle length in monkeys treated with 10 and 100 mg of P. mirifica. Serum levels of follicle stimulating hormone, luteinizing hormone, estradiol, progesterone, or immunoreactive-inhibin did not change during the first and second menstrual cycles of the post-treatment period for all monkey groups. Our findings demonstrate that although changes in hormonal levels could not be observed in this study, a single dose of 1,000 mg of P. mirifica can disturb ovarian function and menstrual cycle in monkeys. PMID- 14745119 TI - Effect of ethanol extracts of three Chinese medicinal plants with anti-diarrheal properties on ion transport of the rat intestinal epithelia. AB - Effects of ethanol extracts of three Chinese medicinal plants, namely, Qinpi (Fraxini cortex), Kushen (Sophora flavescens, AITON), and Huanglian (Coptis teeta, WALLICH), on ion transport of the rat intestinal epithelia were determined in this study. Rat intestinal epithelia mounted in an Ussing chamber attached to a voltage/current clamp were used for measuring changes in the short circuit current across the epithelia. Activation of the intestinal epithelia by serosal administration of 5 microM forskolin resulted in an increase in basal short circuit current. The ethanol extracts of each of the three plants partially reduced the current stimulated by forskolin. In the following experiments, ouabain and bumetanide were added prior to adding the ethanol extract of these plants for revealing their effect on Na(+) and Cl(-) movement. The results suggest that the ethanol extract of the Qinpi would affect Cl(-) transport. On the contrary, the ethanol extract of Kushen would affect Na(+) transport rather than Cl(-) movement. This study provides evidences that reveal the pharmacological mechanism of the Chinese plants with anti-diarrheal properties. PMID- 14745120 TI - Sustained plasma fibrinogen elevation in subtotal nephrectomized rats: effect of cilazapril, an angiotensin-converting enzyme inhibitor. AB - The present study was undertaken to examine whether plasma fibrinogen persistently elevates in subtotal nephrectomized rats, an animal model with inflammatory renal changes. Eight weeks after the induction of 5/6 nephrectomy in male Wistar rats, plasma fibrinogen concentration was determined for the next 12 weeks in the animals received vehicle or an angiotensin-converting enzyme inhibitor, cilazapril (1 or 10 mg/kg per day) orally. In the vehicle-treated nephrectomized rats, plasma fibrinogen concentration significantly (P<0.001) increased (from 127.3 +/- 4.6 [S.E.M.] to 182.3 +/- 5.2 mg/dL) compared with that in the control rats (from 118.0 +/- 2.0 to 153.5 +/- 5.4 mg/dL). Cilazapril attenuated the increases in plasma fibrinogen concentration in a dose-dependent manner. Serum concentration of monocyte chemoattractant protein-1, a key macrophage chemoattractant and activator, increased in the vehicle-treated nephrectomized rats, which was also reduced by cilazapril. These results suggest that plasma fibrinogen elevates persistently in the nephrectomized rats. Local inflammation may be involved in the hepatic fibrinogen synthesis in this model. PMID- 14745121 TI - 5-HT3-receptor antagonist inhibits visceral pain differently in chemical and mechanical stimuli in rats. AB - The present study was designed to compare the effects of a selective 5-HT(3) receptor antagonist, alosetron, on the glycerol-and colorectal distention (CRD) induced visceral nociception as measured by changes in EMG of the external oblique muscle in conscious rats. Both glycerol and CRD evoked the EMG response, and these amplified EMG were attenuated by morphine, indicating that these responses might reflect visceral nociceptive responses. In the present study, we showed that alosetron significantly attenuated the glycerol-induced visceral pain, but not that of CRD. These results suggest that the mechanism of glycerol induced visceral nociception are apparently different from that of CRD. PMID- 14745122 TI - Neuronal nitric oxide synthase is crucial for ganglion cell death in rat retinal explant cultures. AB - We examined possible involvement of nitric oxide synthase (NOS) on ganglion cell death in explant cultures of neonatal rat retina. Survival of retinal ganglion cells was significantly prolonged by a broad-spectrum NOS inhibitor N(omega) nitro-L-arginine methylester. NADPH diaphorase staining revealed a diffused distribution of NOS activity in neuropils of the inner plexiform layer as well as several neurons in the inner nuclear layer. Moreover, 7-nitroindazole but not aminoguanidine promoted the survival of retinal ganglion cells. These results suggest a crucial role of neuronal NOS-derived nitric oxide in retinal ganglion cell death. PMID- 14745123 TI - The inhibitory effects of aqueous extract of Magnolia officinalis on human mesangial cell proliferation by regulation of platelet-derived growth factor-BB and transforming growth factor-beta1 expression. AB - Mesangial cell (MC) proliferation, mediated by platelet-derived growth factor (PDGF)-BB, transforming growth factor (TGF)-beta1, and cyclin-dependent kinases (CDK), is the common feature of glomerulosclerosis. Magnolia officinalis, stem bark of Machilus thunbergii S., has multiple pharmacological effects. In this study, we investigated the influence of aqueous extract of Magnolia officinalis on MC proliferation, DNA synthesis, and expression of PDGF-BB, TGF-beta1, CDK1, CDK2, and CDK4 in fetal bovine serum (FBS)-activated human MC. Magnolia officinalis inhibited the MC proliferation, DNA synthesis, and the expression of PDGF-BB, CDK1, and CDK2 gene and CDK1, CDK2, and TGF-beta1 protein. These results suggest that the inhibitory effect of Magnolia officinalis on MC proliferation may be mediated by regulation of PDGF-BB and TGF-beta1expressions and by modulation of CDK1 and CDK2 expression. PMID- 14745124 TI - [A new paradigm for the progression of advanced heart failure]. AB - To clarify the precise mechanism for the progression of advanced heart failure (AdHF), we assessed the scheme in two HF models, using (I) TO-2 strain hamsters sharing common genetic and clinical features to human families with the delta sarcoglycan (SG) gene mutation and (II) administration of a high-dose (HD) of isoproterenol (Isp) to normal rats. Delta-SG is a component in dystrophin (Dys) related proteins that stabilize the sarcolemma (SL) during repeated heart beats. In TO-2, we followed time course of hemodynamics, immunostaining and Western blotting of Dys and in situ SL permeability by Evans blue uptake with or without the gene therapy. Dys was age-dependently translocated from the SL to myoplasm (MP) where the SL instability accompanied the fragmentation of Dys. By gene therapy to supplement the normal delta-SG gene in hearts in vivo, we found that Dys translocation was selectively improved in cardiomyocytes expressing the delta SG transgene, where the SL fragility was ameliorated. Most importantly, the survival period of the animals was prolonged. Furthermore, Dys but not delta-SG was also time-dependently shifted with a HD of Isp from the SL to MP and fragmented, while delta-SG was preserved intact. We present a novel paradigm that disruption of Dys, but not delta-SG per se, leads to AdHF irrespective of hereditary or acquired origin. PMID- 14745126 TI - [Induction of heat shock protein 70 in failing heart]. AB - Heat shock protein (Hsp) 70s including Hsp72 and Hsp73 are suggested to play an important role in the cardioprotection against stress-induced functional damage. Myocardial tolerance against ischemia/reperfusion injury is increased when myocardial Hsp72 is accumulated after an exposure of normal animals to heat shock. Post-ischemic contractile recovery is improved in the perfused heart of Hsp72-overexpressed mice. However, the role of Hsp72 and Hsp73 in the failing heart following acute myocardial infarction remains unclear. The present study was undertaken to determine whether Hsp72 and Hsp73 production may contribute to the protection of cardiac function in rats with chronic heart failure (CHF) following coronary artery ligation (CAL). The rats with CAL revealed the signs of CHF at the 8th week after the operation. The hearts isolated from rats with CHF were perfused and then subjected to heat shock (at 42 degrees C) for 15 min followed by 6-h perfusion (HS group). The cardiac function of the HS group was markedly decreased and the heat shock-induced increase in myocardial Hsp72 and Hsp73 was attenuated after 6-h perfusion. In the CAL rat treated with the ACE inhibitor trandolapril from the 2nd to the 8th week, induction of Hsp70s was preserved and heat stress-induced reduction in cardiac function was attenuated. The results suggest that a reduction in the production of Hsp70s may play a significant role in the decrease in contractile function during the development of heart failure. PMID- 14745125 TI - [Effects of humoral factors on left ventricular remodeling under chronic heart failure]. AB - Chronic heart failure is a slowly progressive disease. Hemodynamic deterioration activates various neuro-humoral factors and increases stresses, such as catecholamine, angiotensin II (AII), cytokines, endothelin, wall stress, ischemia, tachycardia, and oxidative stress. These factors affect the myocardium to cause phenotype switching, leading to ventricular remodeling. We investigated the effects of pharmacological blocking for neuro-humoral factors in rats with dilated cardiomyopathy. Experimental autoimmune myocarditis (EAM) was elicited in Lewis rats by immunization with cardiac myosin. After acute inflammation healed, rats were treated with angiotensin converting enzyme inhibitors (ACEI), type 1 AII receptor blockers, and amiodarone. These agents had favorable effects on hemodynamics and myocardial contractility, prevented fibrosis, suppressed the expression of ANP, and reversed phenotypic change of cardiac myosin. AII receptor blockers were less effective than ACEI. In order to prevent ventricular remodeling in chronic heart failure, wide and complete blocking of neuro-humoral factors is important. PMID- 14745127 TI - [Energy utility of failing heart]. AB - We investigated left ventricular (LV) mechanoenergetics in acute and chronic failing hearts, induced by high Ca(2+), ischemic-reperfusion injury, diabetes mellitus (DM), and hypothyroidism, using cross-circulated excised rat heart preparations. After high Ca(2+) or ischemic-reperfusion, there was a contractile failure associated with a parallel downward shift of the linear relation between myocardial O(2) consumption per beat (VO(2)) and systolic pressure-volume area (PVA). This result indicated a decrease in VO(2) for total Ca(2+) handling in E-C coupling. We found proteolysis of a cytoskeletal protein, alpha-fodrin. A calpain inhibitor significantly suppressed contractile failure, decreased VO(2) for total Ca(2+) handling, and membrane alpha-fodrin degradation. In DM, the LV relaxation rate was significantly slower, resulting in the decreased O(2) consumption per min for total Ca(2+) handling in E-C coupling. In hypothyroidism, there were systolic and diastolic failures associated with the decreased O(2) consumption per beat for total Ca(2+) handling in E-C coupling. The protein level of sarcoplasmic reticulum Ca(2+) ATPase (SERCA2) was significantly lower in DM and hypothyroidism. We conclude that suppression of O(2) consumption for total Ca(2+) handling, mainly utilized by SERCA2, is a major cause of failing hearts, mediated through degradation of membrane alpha-fodrin via activation of calpain or suppressed expression of SERCA2. PMID- 14745128 TI - [The mechanism of contractile dysfunction in heart failure, focussing on SERCA2a function]. AB - Cytosolic Ca(2+) is a key regulator of excitation-contraction coupling in myocardium. Myocardial contractile dysfunction in heart failure is characterized by a decrease in contraction and prolonged relaxation. These alterations are mainly due to changes in 1) intracellular Ca(2+) transients (CaT), 2) Ca(2+) sensitivity of the contractile elements, and/or 3) contractile proteins. It is useful to investigate the relationship between CaT and contraction for understanding of the mechanism of contractile dysfunction in heart failure. There are many reports regarding the alterations in CaT, Ca(2+) sensitivity, and contractile proteins in heart failure. Changes in the activity of the sarcoplasmic Ca(2+) pump protein, SERCA2a, may be involved in the altered contractility in heart failure. We generated cardiac-restricted overexpression of SERCA2a transgenic mice (TG) and non-transgenic littermates (NTG). To investigate the role of SERCA2a activity for ischemic heart, we used acidosis as a model of acute contractile dysfunction. During acidosis and recovery from acidosis, the peaks of CaT and tension in TG were significantly larger than those in NTG. These results suggest that an increase in the activity of SERCA2a could be beneficial to preserve contractility during acidosis and recovery. Thus, a disturbance of the intracellular Ca(2+) homeostasis is one of the key factors for the contractile dysfunction in heart failure. PMID- 14745129 TI - [Molecular pharmacology of opioid receptors]. AB - We cloned kappa and mu opioid receptor cDNAs. Using these cDNAs, first, we examined the molecular mechanism for the subtype selectivity of opioid ligands, especially a mu-selective ligand DAMGO. Binding experiments using various chimera and mutated receptors revealed that DAMGO discriminates between mu and delta receptors by recognizing the difference in only one amino acid residue, that is, N(127) in mu and K(108) in delta, at the first extracellular loop, and that it distinguishes between mu and kappa receptors by the difference in four amino acid residues at the third extracellular loop. Second, we established the cell lines expressing the cloned mu, delta, or kappa receptor and elucidated the pharmacological properties, that is, binding affinity and agonistic activity of several opioid agonists. Third, distribution of the mRNAs for mu, delta, and kappa receptors in the brain, spinal cord, and DRG was examined by in situ hybridization histochemistry (ISHH). Double ISHH demonstrated that most of the substance P-producing DRG neurons express the micro receptor. Recently, we are interested in the emotional aspect of pain and its regulation by opioids. Behavioral and microdialysis studies showed that sustained pain evoked by the intraplanter injection of formalin induced conditioned place aversion through the increment of glutamate release followed by the activation of NMDA receptors in the basolateral nucleus of amygdala (BLA). Intra-BLA injection of morphine suppressed the place aversion by inhibiting the glutamate release. PMID- 14745130 TI - [The life cycle of the GABA(A) receptor and its regulating molecules]. AB - Gamma-aminobutyric acid(A) (GABA(A)) receptors mediate most of the fast inhibitory neurotransmission in the central nervous system. These ligand-gated ion channels are crucial in the control of cell and network activity. Therefore, modulating their function or cell surface stability will have major consequences for neuronal excitation. This review highlights recent findings on the regulation of GABA(A)-receptor expression and function, focusing on the mechanisms of sorting, targeting, synaptic clustering, and endocytic events of GABA(A) receptors, all which are regulated by their associated proteins. Now these topics are an area of active interest in studies on inhibitory neurotransmission. PMID- 14745131 TI - [Molecular mechanisms of excitatory synaptic transmission: dynamic regulation of AMPA receptors]. AB - AMPA receptors play central roles for synaptic transmission in the central nervous system in mammals. Here we review the molecular mechanisms for how AMPA receptor subunits are correctly assembled, how AMPA receptors are conveyed to dendrites, and how AMPA receptors are exposed in postsynaptic sites. We also discuss the molecular mechanisms of synaptic plasticity that is the cellular basis of learning and memory. PMID- 14745132 TI - Membrane traffic in the post-Golgi network: toward a better understanding of the higher order functioning systems. PMID- 14745133 TI - Vesicle-mediated protein transport pathways to the vacuole in Schizosaccharomyces pombe. AB - The vacuole of Saccharomyces cerevisiae plays essential roles not only for osmoregulation and ion homeostasis but also down-regulation (degradation) of cell surface proteins and protein and organellar turnover. Genetic selections and genome-wide screens in S. cerevisiae have resulted in the identification of a large number of genes required for delivery of proteins to the vacuole. Although the complete genome sequence of the fission yeast Schizosaccharomyces pombe has been reported, there have been few reports on the proteins required for vacuolar protein transport and vacuolar biogenesis in S. pombe. Recent progress in the S. pombe genome project of has revealed that most of the genes required for vacuolar biogenesis and protein transport are conserved between S. pombe and S. cerevisiae. This suggests that the basic machinery of vesicle-mediated protein delivery to the vacuole is conserved between the two yeasts. Identification and characterization of the fission yeast counterparts of the budding yeast Vps and Vps-related proteins have facilitated our understanding of protein transport pathways to the vacuole in S. pombe. This review focuses on the recent advances in vesicle-mediated protein transport to the vacuole in S. pombe. PMID- 14745134 TI - Adaptor protein complexes as the key regulators of protein sorting in the post Golgi network. AB - Adaptor protein (AP) complexes are cytosolic heterotetramers that mediate the sorting of membrane proteins in the secretory and endocytic pathways. AP complexes are involved in the formation of clathrin-coated vesicles (CCVs) by recruiting the scaffold protein, clathrin. AP complexes also play a pivotal role in the cargo selection by recognizing the sorting signals within the cytoplasmic tail of integral membrane proteins. Six distinct AP complexes have been identified. AP-2 mediates endocytosis from the plasma membrane, while AP-1, AP-3 and AP-4 play a role in the endosomal/lysosomal sorting pathways. Moreover, tissue-specific sorting events such as the basolateral sorting in polarized epithelial cells and the biogenesis of specialized organelles including melanosomes and synaptic vesicles are also regulated by members of AP complexes. The application of a variety of methodologies have gradually revealed the physiological role of AP complexes. PMID- 14745135 TI - The structure and function of GGAs, the traffic controllers at the TGN sorting crossroads. AB - GGAs (Golgi-localizing, gamma-adaptin ear homology domain, ARF-binding proteins) are a family of monomeric clathrin adaptor proteins that are conserved from yeasts to humans. Data published during the past four years have provided detailed pictures of the localization, domain organization and structure-function relationships of GGAs. GGAs possess four conserved functional domains, each of which interacts with cargo proteins including mannose 6-phosphate receptors, the small GTPase ARF, clathrin, or accessory proteins including Rabaptin-5 and gamma synergin. Together with or independent of the adaptor protein complex AP-1, GGAs regulate selective transport of cargo proteins, such as mannose 6-phosphate receptors, from the trans-Golgi network to endosomes mediated by clathrin-coated vesicles. PMID- 14745136 TI - The roles of ubiquitin and lipids in protein sorting along the endocytic pathway. AB - After cell surface receptors are internalized for endocytosis, they are accurately sorted in endosomes. Some are recycled to the plasma membrane and others are downregulated by delivery to lysosomes. Evidence is rapidly accumulating that ubiquitination of cargo proteins acts as a sorting signal during endocytosis. Sorting devices that recognize ubiquitin are distributed to various compartments, probably acting in a concerted manner. Cholesterol is enriched in the plasma membrane and endosomes, and is involved in protein sorting by forming microdomains called lipid rafts. Ubiquitin and cholesterol hold the key to control the endocytic sorting, and they are likely acting cooperatively. PMID- 14745137 TI - Lysosome and lysosome-related organelles responsible for specialized functions in higher organisms, with special emphasis on vacuolar-type proton ATPase. AB - Mammals contain various cells differentiated in both morphology and function, which play vital roles in tissue-specific functions. Late endosome/lysosome and lysosomal-related organelles are involved in these specialized functions including antigen presentation, bone remodeling and hormone regulation. To fulfill these diverse roles, lysosomes are present at different levels in different tissues and cell types; however, their morphology within these different tissues varies and the regulation of their activities differs with lysosomal compartments in some cells also functioning as secretory compartments. The luminal acidification of these organelles is closely correlated with their functions. This review will discuss the functions of lysosomes and lysosomal related organelles, with particular emphasis on the major proton pump, the vacuolar-type proton ATPase (V-ATPase), which is responsible for luminal acidification. PMID- 14745138 TI - The roles of Rab27 and its effectors in the regulated secretory pathways. AB - Regulated secretory pathways are highly developed in multicellular organisms as a means of intercellular communication. Each of these pathways harbors unique store organelles, such as granules in endocrine and exocrine tissues and melanosomes in melanocytes. It has recently been shown that the monomeric GTPase Rab27 subfamily regulates the exocytosis of these cell-specific store organelles. Furthermore, genetic alterations of Rab27a cause Griscelli syndrome in humans that manifests as pigmentary dilution of the skin and the hair and variable immunodeficiency due to defects in the transport of melanosomes in melanocytes and lytic granules in cytotoxic T-lymphocytes. Rab27 acts through organelle-specific effector proteins, such as granuphilin in pancreatic beta cells and melanophilin in melanocytes. The Rab27 and effector complex then interacts with proteins that are essential for membrane transport and fusion, such as syntaxin 1a and Munc18-1 for granuphilin and myosin Va for melanophilin. Genome information suggests that other putative Rab27 effector proteins, tentatively termed as exophilins or Slp/Slac2, are predicted to exist because these proteins share the conserved N-terminal Rab27 binding domain and show Rab27-binding activity in vitro or when overexpressed in cell lines. These findings suggest that the Rab27 subfamily regulates various exocytotic pathways using multiple organelle-specific effector proteins. PMID- 14745139 TI - Butyric acid sensitizes Vero cells to ricin-induced apoptosis via accelerated activation of multiple signal transduction pathways. AB - We found that the treatment with 1 mM butyric acid for 2 days renders Vero cells highly sensitive to ricin-induced apoptosis reflected by cytolysis concomitant with apoptotic cellular and nuclear morphological changes, DNA fragmentation, and increase in caspase-3 like activity, whereas butyric acid alone had no cytotoxic effect on Vero cells. During the treatment with butyric acid, gradual increase in alkaline phosphatase activity, an indicator for butyric acid-induced differentiation, was observed in Vero cells. Although the potency of ricin mediated protein synthesis was increased in butyric acid-treated Vero cells as compared to untreated cells, the binding and internalization of ricin to the cells were not much affected. Furthermore, DNA fragmentation caused by other protein synthesis inhibitors such as diphtheria toxin and anisomysin were also highly potentiated in butyric acid-treated Vero cells, whereas the potencies of these toxins to inhibit the protein synthesis were not affected by butyric acid treatment. These results suggest that the apoptosis signaling pathway, which may be triggered by cytotoxic stress response caused by toxins, is sensitized in butyric acid-treated cells, while the pathways leading to the protein synthesis inhibition by these toxins are relatively unchanged. No significant differences in the expression levels of p21, p53, and Bcl-2 proteins were observed between butyric acid-treated and untreated Vero cells. The treatment with ricin resulted in the activation of p38 MAP kinase, and this activation occurred on an accelerated time schedule in butyric acid-treated Vero cells than in untreated cells. The specific inhibitor of p38 MAP kinase SB203580 showed a partial inhibitory effect on ricin-induced apoptosis in control Vero cells, but it was less effective in butyric acid-treated Vero cells. Taken together, our results suggest that butyric acid-treatment may result in sensitization of multiple intracellular signal transduction pathways including apoptotic signaling pathways and p38 MAP kinase pathway. PMID- 14745140 TI - Lipid droplet formation in human myeloid NB4 cells stimulated by all trans retinoic acid and granulocyte colony-stimulating factor: possible involvement of peroxisome proliferator-activated receptor gamma. AB - All trans retinoic acid (ATRA), a differentiation inducer for human myeloid NB4 cells, induced accumulation of lipid droplet as determined by positivity of Nile Red and Oil Red O in this cell line. Granulocyte colony-stimulating factor (G CSF), although not having detectable effect by itself, exerted the additive effects on lipid droplet formation in NB4 cells when combined with ATRA. mRNA analysis for peroxisome proliferator-activated receptors (PPARs) revealed the initial transient downregulation followed by upregulation of the transcript for PPARgamma2, a master molecule for adipogenesis, and upregulation of PPARalpha. BADGE, a synthetic antagonist for PPARgamma, potently inhibited lipid droplet formation in NB4 cells stimulated by ATRA and/or G-CSF, but not the functional differentiation of the cells by ATRA and/or G-CSF. These results suggest that ATRA and G-CSF induce lipid droplet formation via certain PPARgamma-mediated specific mechanisms in human myeloid NB4 cells during functional differentiation. PMID- 14745141 TI - Rationale and design of a large-scale trial using atrial natriuretic peptide (ANP) as an adjunct to percutaneous coronary intervention for ST-segment elevation acute myocardial infarction: Japan-Working groups of acute myocardial infarction for the reduction of Necrotic Damage by ANP (J-WIND-ANP). AB - BACKGROUND: The benefits of percutaneous coronary intervention (PCI) in acute myocardial infarction (AMI) are limited by reperfusion injury. In animal models, atrial natriuretic peptide (ANP) reduces infarct size, so the Japan-Working groups of acute myocardial Infarction for the reduction of Necrotic Damage by ANP (J-WIND-ANP) designed a prospective, randomized, multicenter study, to evaluate whether ANP as an adjunctive therapy for AMI reduces myocardial infarct size and improves regional wall motion. METHODS AND RESULTS: Twenty hospitals in Japan will participate in the J-WIND-ANP study. Patients with AMI who are candidates for PCI are randomly allocated to receive either intravenous ANP or placebo administration. The primary end-points are (1) estimated infarct size (Sigmacreatine kinase and troponin T) and (2) left ventricular function (left ventriculograms). Single nucleotide polymorphisms (SNPs) that may be associated with the function of ANP and susceptibility of AMI will be examined. Furthermore, a data mining method will be used to design the optimal combinational therapy for post-MI patients. CONCLUSIONS: J-WIND-ANP will provide important data on the effects of ANP as an adjunct to PCI for AMI and the SNPs information will open the field of tailor-made therapy. The optimal therapeutic drug combination will also be determined for post-MI patients. PMID- 14745142 TI - Rationale and design of a large-scale trial using nicorandil as an adjunct to percutaneous coronary intervention for ST-segment elevation acute myocardial infarction: Japan-Working groups of acute myocardial infarction for the reduction of Necrotic Damage by a K-ATP channel opener (J-WIND-KATP). AB - BACKGROUND: The benefits of percutaneous coronary intervention (PCI) in acute myocardial infarction (AMI) are limited by reperfusion injury. In animal models, nicorandil, a hybrid of an ATP-sensitive K(+) (KATP) channel opener and nitrates, reduces infarct size, so the Japan-Working groups of acute myocardial Infarction for the reduction of Necrotic Damage by a K-ATP channel opener (J-WIND-KATP) designed a prospective, randomized, multicenter study to evaluate whether nicorandil reduces myocardial infarct size and improves regional wall motion when used as an adjunctive therapy for AMI. METHODS AND RESULTS: Twenty-six hospitals in Japan are participating in the J-WIND-KATP study. Patients with AMI who are candidates for PCI are randomly allocated to receive either intravenous nicorandil or placebo. The primary end-points are (1) estimated infarct size and (2) left ventricular function. Single nucleotide polymorphisms (SNPs) that may be associated with the function of KATP-channel and the susceptibility of AMI to the drug will be examined. Furthermore, a data mining method will be used to design the optimal combined therapy for post-myocardial infarction (MI) patients. CONCLUSIONS: It is intended that J-WIND-KATP will provide important data on the effects of nicorandil as an adjunct to PCI for AMI and that the SNPs information that will open the field of tailor-made therapy. The optimal therapeutic drug combination will also be determined for post-MI patients. PMID- 14745143 TI - More Western hypercholesterolemic patients achieve Japan Atherosclerosis Society LDL-C goals with rosuvastatin therapy than with atorvastatin, pravastatin, or simvastatin therapy. AB - BACKGROUND: Data from Western comparative trials suggest that rosuvastatin is more effective than atorvastatin, simvastatin, and pravastatin in helping hypercholesterolemic patients achieve US and European lipid-lowering guidelines. The purpose of this analysis was to assess the comparative efficacy of rosuvastatin in reducing low-density lipoprotein cholesterol (LDL-C) to levels recommended by the Japan Atherosclerosis Society (JAS). METHODS AND RESULTS: A post hoc analysis of data from 6 randomized, double-blind, active-controlled trials was conducted to evaluate the relative efficacy of rosuvastatin and comparator statins in helping patients achieve the LDL-C goals established by the JAS. The first 5 trials, prospectively designed for pooling, were originally conducted to compare the effects of rosuvastatin with either atorvastatin, simvastatin, or pravastatin in reducing lipid levels and helping patients achieve the LDL-C goals established by the National Cholesterol Education Program. The 6th trial was conducted with similar objectives, but in patients with heterozygous familial hypercholesterolemia (HeFH). Data from 2,139 hypercholesterolemic patients in the first 5 trials were pooled for analysis: rosuvastatin 5 mg (n=390) or 10 mg (n=389) vs atorvastatin 10 mg (n=393); rosuvastatin 5 mg (n=240) or 10 mg (n=226) vs simvastatin 20 mg (n=249) or pravastatin 20 mg (n=252). In the studies with atorvastatin as the comparator, JAS-defined LDL-C goals were reached by 67.2% of the rosuvastatin 5-mg group, 82.3% of the rosuvastatin 10-mg group, and 58.0% of the atorvastatin 10-mg group (p<0.001 for both rosuvastatin groups vs atorvastatin) at 12 weeks. Similarly, in the trials with pravastatin and simvastatin as comparators, the JAS LDL-C goals were reached by 77.5% of the rosuvastatin 5-mg group, 86.7% of the rosuvastatin 10-mg group, 45.2% of the pravastatin 20-mg group and 65.5% of the simvastatin 20 mg group (p<0.001 for both rosuvastatin groups vs pravastatin and simvastatin). In the trial of HeFH patients (n=433 for rosuvastatin, n=187 for atorvastatin), 31.9% of patients treated with rosuvastatin 20 mg achieved JAS LDL-C goals, compared with 17.6% of patients treated with atorvastatin 20 mg (p<0.001). CONCLUSIONS: Rosuvastatin has demonstrated clinical superiority over atorvastatin, pravastatin, and simvastatin in reducing LDL-C levels and in enabling patients to reach goals established by the JAS. PMID- 14745144 TI - Plasma concentrations of monocyte chemoattractant protein 1 (MCP-1) and neopterin in the coronary circulation of patients with coronary artery disease. AB - BACKGROUND: The dynamics of MCP-1 and neopterin and the relation between their concentrations in coronary circulation and the severity of coronary atherosclerosis were evaluated in patients with stable coronary artery disease (CAD). METHODS AND RESULTS: Blood samples were obtained from the aortic root (Ao) and coronary sinus (CS) of 78 patients who underwent coronary angiography. Plasma MCP-1 and neopterin concentrations were measured using an enzyme-linked immunosorbent assay method and the CS-Ao differences were calculated. The severity of coronary heart disease (CHD) was evaluated in 52 patients who had no history of coronary angioplasty, using 3 coronary scoring systems: the clinical 1 to 3-vessel disease score, the American Heart Association extension score (1-15 segments), and the Gensini score. The plasma MCP-1 and neopterin concentrations increased significantly with age. The CS-Ao differences for neopterin showed weak, but significant, positive correlation with the Gensini score (r=0.347, p=0.013). There were no correlations among the MCP-1 concentrations in the Ao or CS, or in the CS-Ao difference, with the severity of CHD. CONCLUSIONS: The results indicate that neopterin is a useful marker of the severity of coronary atherosclerosis in patients with stable CAD, acting as an index of the activity of monocytes/macrophages. PMID- 14745145 TI - Statins induce the regression of left ventricular mass in patients with angina. AB - Background There is evidence that statins induce the regression of cardiac hypertrophy in a transgenic rabbit model of hypertrophic cardiomyopathy. Methods and Results The association between treatment with statins and the regression of cardiac mass (left ventricular mass index, LVMI) was investigated in a case - control study using transthoracic echocardiography in 304 patients with angina who underwent coronary angiography. Those who received pravastatin or simvastatin were defined as cases (n=66), and age, sex and body mass index-matched controls (n=127) were selected. The cases showed a significant decrease in LVMI compared with the controls. Although the cases included a significantly higher percentage of patients with hypertension and calcium antagonist (CaA) treatment than the controls, there were no relationships between LVMI and either hypertension or CaA treatment. Because the cases had a significantly higher number of stenosed vessels than the controls, LVMI for each number of stenosed vessels was analyzed, and a significant interaction effect between the association of LVMI with statin and the number of stenosed vessels was found. Conclusions Treatment with statins was associated with a lower cardiac mass in patients with angina, suggesting that this is one of the drugs' pleiotropic effects. PMID- 14745146 TI - Arrhythmias late after repair of tetralogy of fallot: a Japanese Multicenter Study. AB - BACKGROUND: Arrhythmia is a major late complication in adults with repaired tetralogy of Fallot, although a large-scale study has not been carried out in Japan. METHODS AND RESULTS: A nationwide multicenter study with 512 operative survivors was performed. Actuarial survival rate at 30 years (maximum follow-up) was 98.4%. Fifty-four patients (10.5%) had clinically important arrhythmias, including 23 patients with bradycardia caused by sick sinus syndrome or atrioventricular block (AVB). A patient with complete AVB (CAVB) without pacemaker implantation (PMI) died later. Two patients had sustained ventricular tachycardia (VT) and syncope was reported in 18 patients with ventricular arrhythmias (VA). Atrial tachyarrhythmias were observed in 13 patients. Older age at operation was a risk factor for atrial fibrillation/flutter, longer postoperative survival duration for VA, and QRS duration >120 ms for VT. Perimembranous ventricular septal defect was related to CAVB. Right ventricular outflow patch was not a risk factor. Importantly, 60% of the subjects had QRS duration <120 ms. CONCLUSION: The prevalence of serious arrhythmias is low in Japanese TOF patients as compared with the results from Western countries. CAVB without PMI and VT are the major risk factors for late morbidity and mortality. The excellent result could be related to the narrow QRS after surgery. PMID- 14745147 TI - Double-counting of intracardiac electrogram during biatrial pacing. AB - BACKGROUND: Biatrial pacing has a significant problem with memory function that misinterprets normal sinus rhythm as atrial tachyarrhythmias and in addition estimation of the atrial pacing thresholds (biatrial and uniatrial pacing thresholds) is sometimes difficult because of small P waves. METHODS AND RESULTS: The intracardiac electrograms recorded by a pacemaker in 10 patients (age, 66.7+/ 10.7 (SD) years) with implanted biatrial pacemakers were analyzed. Atrial sensing within the atrial refractory period after atrial pacing was counted in 6 of the 10 patients (timing of the double counting was 143+/-64 ms) when pacing failed in the left or right atrium. Atrial sensing within the atrial refractory period after atrial pacing disappeared when biatrial pacing was successfully performed. Atrial double-counts depend on interatrial conduction delay. The memory function of implanted pacemaker devices misinterpreted normal sinus rhythm as atrial tachyarrhythmias because of atrial double-counts. On the other hand, the biatrial pacing threshold was easily recognized using this phenomenon. CONCLUSIONS: The memory function of pacemaker devices is unreliable because of atrial double counting during sinus rhythm in patients with biatrial pacing. However, the biatrial pacing threshold is easily checked using this phenomenon. PMID- 14745148 TI - Evaluation of the usefulness of recording the ECG in the 3rd intercostal space and prevalence of Brugada-type ECG in accordance with recently established electrocardiographic criteria. AB - BACKGROUND: It has been reported that recording electrocardiograms (ECGs) in the 3rd intercostal space (ICS) is one method that can be used for detecting Brugada syndrome; however, the prevalence of Brugada-type ECGs recorded in the 3rd ICS and the usefulness of recording the ECG in the 3rd ICS in accordance with recently established electrocardiographic criteria is unknown. METHODS AND RESULTS: ECGs were recorded in both the 4th and 3rd ICS in 17 Brugada-type ECG patients (group A) and in 206 consecutive male subjects (group B). Brugada-type ECGs were divided into 3 types. In group A, the prevalence of type 1 ECG, which is a coved-type ECG with ST-segment elevation of >/=2 mm, increased from 23.5% to 64.7% when ECG was recorded in the 3rd ICS. The conversion to type 1 ECG was found to be related to induction of ventricular arrhythmia. In group B, the prevalence of Brugada-type ECG increased from 1.5% to 5.8% when the ECG was recorded in the 3rd ICS. CONCLUSIONS: Recording the ECG in the 3rd ICS is useful for identifying high-risk patients with Brugada-type ECG and for detecting concealed Brugada-type ECG. PMID- 14745149 TI - Severity indices of right coronary cusp prolapse and aortic regurgitation complicating ventricular septal defect in the outlet septum: which defect should be closed? AB - BACKGROUND: The factors that may determine the evolution of right coronary cusp prolapse (RCCP) and regurgitation (AR) associated with a ventricular septal defect in the outlet septum (outlet VSD) have not been clarified. METHODS AND RESULTS: The Doppler echocardiograms of 316 patients were grouped according to both the development of RCCP, and the values of the right coronary cusp deformity index (RCCD) and the right coronary cusp imbalance index (R/L). All detected AR was /=0.30 or R/L >/=0.30. Rupture of the sinus of Valsalva was identified in patients with RCCD >/=0.30. A significantly large number of patients with both RCCD >/=0.30 and R/L >/=1.30 (p<0.01), and a few patients with either RCCD >/=0.30 or R/L >/=0.30 underwent aortic valvuloplasty or replacement. Operative outcome for AR /=0.30 or R/L >/=1.30. PMID- 14745150 TI - Office cardiologists cooperative study on whole blood rapid panel tests in patients with suspicious acute myocardial infarction: comparison between heart type fatty acid-binding protein and troponin T tests. AB - BACKGROUND: The whole blood rapid troponin T test, used to determine the early diagnosis of acute myocardial infarction (AMI), is effective only for 3-4 h after onset. METHODS AND RESULTS: The present office cardiologists cooperative study compared the diagnostic efficacy of a newly developed whole blood rapid panel test for heart-type fatty acid-binding protein (H-FABP) with that of the rapid troponin T test in 129 consecutive patients with suspicious AMI according to certain time-frames from onset to presentation. Thirty-one patients (24.0%) had a final diagnosis of AMI. The respective sensitivities of the rapid H-FABP and troponin T tests were 100% vs 50% (p<0.05) for patients presenting within 3 h of onset; 75% vs 0% for those between 3 and 6 h; 100% vs 60% for those between 6 and 12 h; and 100% vs 100% for those presenting later than 12 h. The respective specificities were 63% vs 96.3% (p<0.05); 93.8% vs 93.8%; 72.7% vs 100%; and 75.0% vs 87.5%. Negative predictive value was 100% vs 86.7%; 93.8% vs 78.9%; 100% vs 84.6%; and 100% vs 100%, respectively. Patients with non-AMI myocardial damage associated with unstable angina or severe heart failure showed positive H-FABP test results and blunted the specificity. CONCLUSIONS: When using the novel rapid H-FABP test, cardiac emergency triage to exclude non-AMI patients should be effectively organized within 3 h of onset. PMID- 14745151 TI - Monomethoxypolyethylene glycol-modified cardiac myosin treatment blocks the active and passive induction of experimental autoimmune myocarditis. AB - BACKGROUND: Injecting various protein antigens conjugated to monomethoxypolyethylene glycol (mPEG) results in antigen-specific tolerance to subsequent immunization. In the present study the ability of mPEG-modified cardiac myosin (CM) to block the development of experimental autoimmune myocarditis (EAM) induced by CM immunization or by the transfer of lymphocytes from CM-immunized donors was studied. METHODS AND RESULTS: A/J mice were injected with mPEG-CM before active or passive EAM induction. We examined the suppressive mechanism by the transfer of lymphocytes from mPEG-CM-treated mice into naive mice. To ascertain the cells responsible for suppressing EAM induction, in vivo or in vitro depletion of CD4(+) or CD8(+) T cells was performed. mPEG-CM administered before active or passive EAM induction markedly suppressed the incidence and severity of EAM and reduced CM-specific antibody responses. When lymphocytes from mPEG-CM treated mice were transferred into naive mice that were then immunized with CM, the suppressive effect was recapitulated. CONCLUSIONS: mPEG-CM treatment blocked the active and passive induction of EAM. PMID- 14745152 TI - Difference in the cardioprotective mechanisms between ischemic preconditioning and pharmacological preconditioning by diazoxide in rat hearts. AB - BACKGROUND: Recent studies have implicated the opening of mitochondrial K(ATP) (mitoK(ATP)) channels and the production of reactive oxygen species (ROS) in the cardioprotective mechanism of ischemic preconditioning (IPC). METHODS AND RESULTS: The involvement of mitoK(ATP) channels and ROS in the cardioprotective effects of both IPC and the mitoK(ATP) channel opener diazoxide (DZ) was investigated in ischemic/reperfused rat hearts. The effects of IPC and DZ on myocardial high-energy phosphate concentrations and intracellular pH (pH(i)) were also examined using (31)P nuclear magnetic resonance spectroscopy. Although both the mitoK(ATP) channel inhibitor 5-hydroxydecanoate and the antioxidant N acetylcysteine abolished the postischemic recovery of contractile function by DZ, neither of them inhibited that by IPC. IPC attenuated the decline in pHi during ischemia, but DZ did not (6.28+/-0.04 in IPC, p<0.05, and 6.02+/-0.05 in DZ vs 6.02 +/-0.06 in control hearts). DZ, but not IPC, reduced the decrease in ATP levels during ischemia (ATP levels at 20-min ischemia: 26.3+/-3.4% of initial value in DZ, p<0.05, and 8.1+/-3.0% in IPC vs 15.1+/-1.3% in control hearts). CONCLUSIONS: These results suggest that DZ-induced cardioprotection is related to ROS production and reduced ATP degradation during ischemia, whereas attenuated acidification during ischemia is involved in IPC-induced cardioprotection, which is not mediated through mitoK(ATP) channel opening or ROS production. PMID- 14745153 TI - Troponin-T and brain natriuretic peptide as predictors for adriamycin-induced cardiomyopathy in rats. AB - BACKGROUND: Clinical methods for the early detection of doxorubicine (adriamycin; ADR) -induced cardiotoxicity have not been established. This study prospectively investigated whether atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP) and cardiac troponin T (TnT) are predictors for ADR-induced cardiotoxicity, and examined the correlations between the serum concentrations of these biomarkers and the functional alternations associated with ADR-induced myocardial damage. METHODS AND RESULTS: Male Wistar rats were injected weekly with 2 mg/kg of ADR via the tail vein for 8 weeks to induce cardiotoxicity. Echocardiograms of each ether anesthetized rat were taken at 6, 8, 10 and 12 weeks after the first administration of ADR, and blood samples collected from the tail vein were used to quantify plasma ANP and BNP, and serum TnT after echocardiography. Plasma BNP and serum TnT significantly increased from 6 to 12 weeks (81.5 to 173.3 pg/ml (p<0.001), <0.01 to 1.09 ng/ml (p<0.05), respectively) with deterioration of left ventricular % fractional shortening (%FS) (58.6% to 36.8%). The %FS significantly correlated with TnT (r=-0.51, p<0.001) and BNP (r=-0.75, p<0.0001); however, the increase of TnT was antecedent to the increase of BNP and the deterioration of %FS. CONCLUSION: Plasma BNP and serum TnT concentrations, especially TnT, measured by this highly sensitive method are useful predictors for ADR-induced cardiomyopathy. PMID- 14745154 TI - Complex atrial reentrant circuits evaluated by entrainment mapping using a multielectrode basket catheter. AB - Atrial tachycardias after open heart surgery sometimes have complex reentrant circuits. A patient with a dual-loop atrial reentrant circuit occurring after mitral valve replacement was evaluated by entrainment mapping with a basket catheter. The position of the catheter was adjusted to obtain atrial electrograms of the anterior and posterior septal areas, the crista terminalis, the free wall, and the tricuspid annular region. Entrainment mapping identified a dual-loop reentry consisting of one circuit around the tricuspid annulus and another around the septal atriotomy scar. The reentrant circuit around the septal incision was eliminated by ablating the area between the septal incision and the inferior vena cava, and the circuit around the tricuspid annulus was terminated with an additional linear ablation between the tricuspid annulus and the inferior vena cava. Entrainment mapping using a multielectrode basket catheter is very useful for identifying complex atrial reentrant circuits. PMID- 14745155 TI - Successful treatment of primary cardiac lymphoma with monoclonal CD20 antibody (rituximab). AB - Primary cardiac malignant lymphoma is extremely rare and almost all patients die within weeks. Monoclonal CD20 antibody (rituximab) was administered to a patient with primary cardiac B-cell non-Hodgkin's lymphoma expressing a CD20 molecule. The results suggest that rituximab may be a safe and effective new therapy for primary cardiac B-cell lymphoma. PMID- 14745156 TI - 17-year follow-up study of a patient with obstructive hypertrophic cardiomyopathy with a deletion mutation in the cardiac myosin binding protein C gene. AB - A 60-year-old Japanese man with obstructive hypertrophic cardiomyopathy was found to have a mutation in the cardiac myosin binding protein C gene: a single base deletion of a thymidine residue at nucleotide 11645 (codon 593) in exon 18. He was diagnosed at the age of 43 and has been followed for 17 years. During this follow-up period, echocardiograms and mechanocardiograms revealed progressive hypertrophy until the age of 54, then gradual dilation of the left ventricle associated with a decrease in the obstruction. Paroxysmal atrial fibrillation occurred at the age of 52 and progressed to chronic atrial fibrillation at the age of 53. He had congestive heart failure at the age of 58. PMID- 14745157 TI - Cyanotic tetralogy of fallot with its infective endocarditis complication on the tricuspid and pulmonary. AB - A 55-year-old man had undiagnosed tetralogy of Fallot with the complications of decompensated heart failure and infective endocarditis, as well as pulmonic involvement secondary to the endocarditis. The patient had a massive hemoptysis and died. This case is a rare insight into the late outcome of this congenital heart disease. PMID- 14745158 TI - Molecular dissection of the Selenomonas ruminantium cell envelope and lysine decarboxylase involved in the biosynthesis of a polyamine covalently linked to the cell wall peptidoglycan layer. AB - The wild type of Selenomonas ruminantium subsp. lactilytica, which is a strictly anaerobic, Gram-negative bacterium isolated from sheep rumen, requires one of the normal saturated volatile fatty acids with 3 to 10 carbon atoms for its growth in a glucose medium; however, no such obligate requirement of fatty acid is observed when the cells are grown in a lactate medium. This bacterium is characterized by a unique structure of the cell envelope and a novel lysine decarboxylase and its regulatory protein. In the first part of this article, we will refer to the chemical structure of phospholipid and lipopolysaccharide in the cell membranes of this bacterium compared with that from the general Gram-negative bacteria for understanding their biological functions. S. ruminantium has neither free nor bound forms of Braun lipoprotein which plays an important role of the maintenance of the structural integrity of the cell surface in general Gram-negative bacteria. However, S. ruminantium has cadaverine, which links covalently to the peptidoglycan as a pivotal constituent for the cell division. In the second part of this article, we will refer to the chemical structure of the cadaverine containing peptidoglycan, its biosynthesis, and the biological function. In the third part of this article, we will depict the molecular cloning of the genes encoding S. ruminanitum lysine decarboxylase (LDC) and its regulatory protein of 22-kDa (22-kDa protein; P22) which has similar characteristics to that of antizyme of ornithine decarboxylase in eukaryotic cells, and the molecular dissection of these proteins for understanding the regulation of cadaverine biosynthesis. Finally, we will illustrate a proposed structure of the cell envelope, a processes of biosynthesis of the cadaverine-containing peptidoglycan layer, and the LDC degradation mechanism in S. ruminantium, on the basis of the analyses of the cell envelope components, the results from the in vitro experiments on the biosynthesis of the peptidoglycan layer, and the current status of the knowledge on LDC and P22 in this organism. PMID- 14745159 TI - Distribution of Prx-linked hydroperoxide reductase activity among microorganisms. AB - Peroxiredoxin (Prx) constitutes a large family of enzymes found in microorganisms, animals, and plants, but the detection of the activities of Prx linked hydroperoxide reductases (peroxiredoxin reductases) in cell extracts, and the purification based on peroxide reductase activity, have only been done in bacteria and Trypanosomatidae. A peroxiredoxin reductase (NADH oxidase) from a bacterium, Amphibacillus, displayed only poor activities in the presence of purified Prx from Saccharomyces or Synechocystis, while it is highly active in the presence of bacterial Prx. These results suggested that an enzyme system different from that in bacteria might exist for the reduction of Prx in yeast and cyanobacteria. Prx-linked hydroperoxide reductase activities were detected in cell extracts of Saccharomyces, Synechocystis, and Chlorella, and the enzyme activities of Saccharomyces and Chlorella were induced under vigorously aerated culture conditions and intensive light exposure conditions, respectively. Partial purification of Prx-linked peroxidase from the induced yeast cells indicated that the Prx-linked peroxidase system consists of two protein components, namely, thioredoxin and thioredoxin reductase. This finding is consistent with the previous report on its purification based on its protein protection activity against oxidation [Chae et al., J. Biol. Chem., 269, 27670-27678 (1994)]. In this study we have confirmed that Prx-linked peroxidase activity are widely distributed, not only in bacteria species and Trypanosomatidae, but also in yeast and photosynthetic microorganisms, and showed reconstitution of the activity from partially purified interspecies components. PMID- 14745160 TI - Rare bacterium of new genus isolated with prolonged enrichment culture. AB - Dynamic change in microbial flora was monitored with an oxygen electrode. The 1st phase microorganisms, which first grew well in LB medium, were followed by the 2nd phase microorganisms, which supposedly assimilated microbial cells of the 1st phase and their metabolites. In a similar way, a change in microbial flora was observed from the 1st phase to the 4th phase in 84 hr. Based on this observation, prolonged enrichment culture was done for as long as two months to increase the ratio of existence of rare microorganisms. From these culture liquids, four slow growing bacteria (provisionally named Shinshu-ah1, -ah2, -ah3, and -ah4), which formed scarcely visible small colonies, were isolated. Sequence analysis of their 16S rDNA showed that Shinshu-ah1 had 97% homology with Bradyrhizobium japonicum and uncultured alpha proteobacterium clone blaii 16, Shinshu-ah2 91% with Rasbo bacterium, Alpha proteobacterium 34619, Bradyrhizobium genosp. P, Afipia felis and an unidentified bacterium, Shinshu-ah3 99% with Methylobacterium mesophilicum, and Shinshu-ah4 95% with Agromyces ramosus DSM 43045. Phylogenetic study indicated that Shinshu-ah2 had a possibility to form a new family, Shinshu ah1 a new genus, and Shinshu-ah4 a new species. PMID- 14745161 TI - Antioxidative effects of glycosyl-ascorbic acids synthesized by maltogenic amylase to reduce lipid oxidation and volatiles production in cooked chicken meat. AB - Glycosylated ascorbic acids were synthesized by using the transglycosylation activity of Bacillus stearothermophilus maltogenic amylase with maltotriose to show effective antioxidative activity with enhanced oxidative stability. The modified ascorbic acids comprised mono- and di-glycosyl transfer products with an alpha-(1,6)-glycosidic linkage. The antioxidative effects of the glycosyl derivatives of ascorbic acid on the lipid oxidation of cooked chicken breast meat patties were compared, and the synergistic effect when combined with alpha tocopherol was determined in terms of thiobarbituric acid-reactive substances (TBARS) and volatiles production during storage. The results indicate that the glycosylated ascorbic acids had very effective antioxidative activity in preventing lipid oxidation, and were better in their synergistic effect in comparison to authentic ascorbic acid, with maltosyl-ascorbic acid being the most effective. Volatiles production was highly correlated with the TBARS values in the lipid oxidation of cooked meat. The antioxidative effect preventing the production of volatiles was particularly strong on pentanal, fairly strong on propanal and butanal, and not at all on ethanal. Propanal, pentanal, and the total volatiles thus provided a good representation of the lipid oxidation status of cooked chicken meat. PMID- 14745162 TI - Effects of excess nicotinamide administration on the urinary excretion of nicotinamide N-oxide and nicotinuric acid by rats. AB - We investigated a useful chemical index for an excessive nicotinamide intake and how this excessive nicotinamide intake affects the tryptophan-nicotinamide metabolism in rats. Weaning rats were fed on a tryptophan-limited and nicotinic acid-free diet containing no, 0.003%, 0.1%, 0.2%, or 0.3% nicotinamide for 21 days. Urine samples were collected on the last day and analyzed the intermediates and metabolites on the tryptophan-nicotinamide pathway. Nicotinamide N-oxide, nicotinic acid and nicotinuric acid, metabolites of nicotinamide, were detected when nicotinamide at more than 0.1% had been taken. An intake of nicotinamide of more than 0.1% increased the urinary excretion of quinolinic acid, an intermediate on the pathway. Nicotinamide N-oxide and nicotinuric acid increased with increasing dietary concentration of nicotinamide. These results show that the measurements of nicotinamide N-oxide and nicotinuric acid in urine would be useful indices for an excessive nicotinamide intake. PMID- 14745163 TI - Simultaneous determination of isoflavones and bisphenol A in rat serum by high performance liquid chromatography coupled with coulometric array detection. AB - A method for simultaneous detection and quantification is presented to determine the presence of isoflavones and bisphenol A in a biological sample. A coulometric array detector was used with reversed-phase high-performance liquid chromatography (HPLC). Daidzein (1), glycitein (2), genistein (3) and their glucoside conjugates, daidzin (4), glycitin (5) and genistin (6), were measured as phytochemicals. Also assayed here was equol (7), a metabolite from compound 1, and bisphenol A (8), an industrial chemical that acts as an endocrine disrupter. All chemicals were simultaneously detected by using a 600-mV single detection voltage with high efficacy. A mixture of 1, 3 and 8 was orally administered to rats, and the levels of these three chemicals in the serum were clearly increased after a 4 kU beta-glucuronidase treatment. The levels of compounds 1 and 3 in the serum were detected at 1665 and 2040 ng/ml, while 8 was at a low level of 417 ng/ml. Compound 7 in the serum was not detected until after enzymatic hydrolysis (72 ng/ml). These results suggest that this analytical method would be useful for metabolic and pharmacokinetic studies on isoflavones and bisphenol A. PMID- 14745164 TI - Isolation of tryptophan as an inhibitor of ovalbumin permeation and analysis of its suppressive effect on oral sensitization. AB - Tryptophan was isolated from rat feces as an active compound against ovalbumin permeation in an in vitro Caco-2 cell model. Tryptophan dose-dependently inhibited ovalbumin permeation with accompanying increase in transepithelial electric resistance, and its inhibitory activity reached a plateau at 10 mM. Brown Norway rats were sensitized by intragastric administration of ovalbumin together with or without tryptophan. Antibody levels specific to ovalbumin in the sera and proliferative responses of spleen mononuclear cells to ovalbumin were significantly lower in rats administered ovalbumin plus tryptophan than those administered ovalbumin alone. These results suggest that tryptophan suppresses oral sensitization to ovalbumin, probably via suppression of ovalbumin absorption from the intestinal tract. PMID- 14745165 TI - Encapsulation of shiitake (Lenthinus edodes) flavors by spray drying. AB - Powdery encapsulation of shiitake flavors, extracted from dried shiitake, was investigated by spray drying. Flavor retention increased with an increase in drying air temperature and solid content, and decreased with an increase in dextrose equivalents of maltodextrin. A heat-treatment of the extract liquid made the lenthionine concentration increase, but did not influence the concentrations of the other flavors. The formation of lenthionine with heat-treatment could be described by the consecutive unimolecular-type first order reaction. Lenthionine content in a spray-dried powder prepared with the heated extracted liquid significantly increased. alpha-Cyclodextrin was the most suitable encapsulant of alpha-, beta-, and gamma-cyclodextrins to prepare the spray-dried powder, including lenthionine. The flavor retentions were markedly increased by using of alpha-cyclodextrin and maltodextrin in combination as an encapsulant. PMID- 14745166 TI - Comparison of the changes in lipid metabolism between hepatoma-bearing and lipopolysaccharide-treated rats. AB - To elucidate the mechanism for hyperlipidemia in the hepatoma-bearing state, changes in some parameters related to the lipid metabolism and serum tumor necrosis factor-alpha (TNF-alpha) level were examined in Donryu rats that had been subcutaneously implanted with an ascites hepatoma cell line of AH109A. These parameters were also examined in rats that had been given a single injection of lipopolysaccharide (LPS), a model for acute infection with TNF-alpha secretion into the blood circulation. The serum triglyceride and total cholesterol (Ch) levels were significantly higher in both the hepatoma-implanted and LPS-injected rats than in normal rats. The level of adipose tissue lipoprotein lipase was decreased by hepatoma implantation and LPS injection, while the hormone-sensitive lipase activity was increased by the same treatments. Fatty acid (FA) oxidation and Ch synthesis were also stimulated by both treatments. The serum TNF-alpha level was noticably elevated by hepatoma implantation and greatly by the LPS injection. This LPS injection increased hepatic FA synthesis. The serum high density lipoprotein Ch level and hepatic Ch 7alpha-hydroxylase activity were not changed by the LPS injection. Hepatoma implantation led to hyperlipidemia and elevated the serum TNF-alpha level, as did the LPS injection. PMID- 14745167 TI - Catalytic Reaction of Basidiomycete Lentinula edodes Cytochrome P450, Le.CYP1 Enzyme Produced in Yeast. AB - The basidiomycete Lentinula edodes (Le.) cytochrome P450, Le.CYP1 was functionally expressed in Saccharomyces cerevisiae. The microsomal fraction containing Le.CYP1 was prepared from the recombinant yeast and the Le.CYP1 was analyzed. The 7-ethoxycoumarin and benzo(a)pyrene were found to be the substrates of Le.CYP1 enzyme. Le.CYP1 converted 7-ethoxycoumarin to 7-hydroxycoumarin. PMID- 14745168 TI - Triterpene acids from the leaves of Perilla frutescens and their anti inflammatory and antitumor-promoting effects. AB - Nine triterpene acids, viz., six of the ursane type, ursolic acid (1), corosolic acid (2), 3-epicorosolic acid (3), pomolic acid (4), tormentic acid (5) and hyptadienic acid (6), and three of the oleanane type, oleanolic acid (7), augustic acid (8) and 3-epimaslinic acid (9), among which 1 constituted the most predominant triterpene acid, were isolated and identified from ethanol extracts of the leaves of red perilla [Perilla frutescens (L.) Britton var. acuta Kudo] and green perilla [P. frutescens (L.) Britton var. acuta Kudo forma viridis Makino]. These eight compounds, 1, 2, 4-9, were evaluated for their inhibitory effects on 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced inflammation (1 microg/ear) in mice. All the compounds tested showed a marked anti-inflammatory effect, with a 50% inhibitory dose (ID50) of 0.09-0.3 mg per ear. In addition, an evaluation against the Epstein-Barr virus early antigen (EBV-EA) activation induced by TPA showed five compounds, 1-3, 5 and 9, with a potent inhibitory effect on EBV-EA induction (91-93% inhibition at 1x10(3) mol ratio/TPA). Furthermore, compound 5 exhibited strong antitumor-promoting activity in an in vivo two-stage carcinogenesis test of mouse tumor by using 7,12 dimethylbenz(a)anthracene (DMBA) as an initiator and TPA as a promoter. PMID- 14745169 TI - Decomposition kinetics of maltose in subcritical water. AB - The decomposition process of maltose in subcritical water was studied using a tubular reactor in the temperature range of 180 to 260 degrees C and at 10 MPa. The formation of glucose and 5-hydroxymethyl-2-furaldehyde during the maltose decomposition was also observed. The decomposition rate of maltose was faster at higher temperatures. The rate was approximated by first-order kinetics during the early stage of the decomposition, but was accelerated and deviated from these kinetics at the later stage. The effluent pH decreased as the residence time in the reactor increased and the decrease of pH affected the maltose decomposition rate and glucose formation. Low pH of a feed solution accelerated maltose decomposition. A good correlation was obtained between the pH of the effluent and the rate constant of the first-order kinetics. PMID- 14745170 TI - High level expression of thermostable lipase from Geobacillus sp. strain T1. AB - A thermostable extracellular lipase of Geobacillus sp. strain T1 was cloned in a prokaryotic system. Sequence analysis revealed an open reading frame of 1,251 bp in length which codes for a polypeptide of 416 amino acid residues. The polypeptide was composed of a signal peptide (28 amino acids) and a mature protein of 388 amino acids. Instead of Gly, Ala was substituted as the first residue of the conserved pentapeptide Gly-X-Ser-X-Gly. Successful gene expression was obtained with pBAD, pRSET, pET, and pGEX as under the control of araBAD, T7, T7 lac, and tac promoters, respectively. Among them, pGEX had a specific activity of 30.19 Umg(-1) which corresponds to 2927.15 Ug(-1) of wet cells after optimization. The recombinant lipase had an optimum temperature and pH of 65 degrees C and pH 9, respectively. It was stable up to 65 degrees C at pH 7 and active over a wide pH range (pH 6-11). This study presents a rapid cloning and overexpression, aimed at improving the enzyme yield for successful industrial application. PMID- 14745171 TI - Tetrapetalone A, a novel lipoxygenase inhibitor from Streptomyces sp. AB - A simple new assay was designed for lipoxygenase inhibitors. This assay was used to find the novel lipoxygenase inhibitor, tetrapetalone A (1). Tetrapetalone A (1), C26H33NO7, was isolated from Streptomyces sp. USF-4727 strain. Its planar structure was determined by spectroscopic evidence and by methylating with diazomethane to show the presence of a novel tetracyclic skeleton and a beta-D rhodinosyl moiety. The stereochemistry of 1 was investigated by the coupling constant in the 1H-NMR spectrum, NOE correlations, modified Mosher's method and derivation. We have reported the structural elucidation of 1 in our previous paper. However, further investigation gave another structure for 1, which is described in this paper. Tetrapetalone A showed similar inhibitory activity against soybean lipoxygenase to the two well-known lipoxygenase inhibitors, kojic acid and NDGA, while methylated tetrapetalone A (2) showed little inhibitory activity, even at a concentration of 1 mM. PMID- 14745172 TI - Proteomic identification of alpha-amylase isoforms encoded by RAmy3B/3C from germinating rice seeds. AB - We isolated and identified 10 alpha-amylase isoforms by using beta-cyclodextrin Sepharose affinity column chromatography and two-dimensional polyacrylamide gel electrophoresis from germinating rice (Oryza sativa L.) seeds. Immunoblots with anti-alpha-amylase I-1 and II-4 antibodies indicated that 8 isoforms in 10 are distinguishable from alpha-amylase I-1 and II-4. Peptide mass fingerprinting analysis showed that there exist novel isoforms encoded by RAmy3B and RAmy3C genes. The optimum temperature for enzyme reaction of the RAmy3B and RAmy3C coding isoforms resembled that of alpha-amylase isoform II-4 (RAmy3D). Furthermore, complex protein polymorphism resulted from a single alpha-amylase gene was found to occur not only in RAmy3D, but also in RAmy3B. PMID- 14745174 TI - Anisodamine causes the changes of structure and function in the transmembrane domain of the Ca(2+)-ATPase from sarcoplasmic reticulum. AB - The effects of anisodamine on the Ca(2+)-ATPsae of sarcoplasmic reticulum (SR) were investigated by using differential scanning calorimetry to measure the ability of anisodamine to denature the transmembrane domain and the cytoplasmic domain. Anisodamine significantly altered the thermotropic phase behaviors of the transmembrane domain of purified Ca(2+)-ATPase. Specifically, the melting temperature of the transmembrane domain moved toward lower temperatures with the concentrations of anisodamine increasing and the thermotropic phase peak was abolished at 10 mM, indicating that the stabilized structure of the transmembrane domain in the presence of Ca2+ could be destabilized by anisodamine. Decreases of the intrinsic fluorescence and increases of the extrinsic fluorescence of ANS, a fluorescent probe, showed the exposure of tryptophan and hydrophobic region, respectively, suggesting again that anisodamine caused a less compact conformation in the transmembrane domain. A marked inhibition of the Ca2+ uptake activity of SR Ca(2+)-ATPase was observed when the addition of anisodamine. The drug did not affect the cytoplasmic domain of the enzyme and only slightly decreased the ATPase activity of the enzyme at concentrations up to 10 mM. This was likely due to the destabilized protein transmembrane domain. To sum up, our results revealed that anisodamine interacted specifically with the transmembrane domain of SR Ca(2+)-ATPase and inhibited the Ca2+ uptake activity of the enzyme. PMID- 14745173 TI - A hydroxyl group of flavonoids affects oral anti-inflammatory activity and inhibition of systemic tumor necrosis factor-alpha production. AB - We previously reported that oral administration of luteolin can inhibit serum tumor necrosis factor (TNF)-alpha production and several inflammatory and allergic models. We investigated here the effect of various flavonoids which resemble luteolin in structure. Lipopolysaccharide (LPS)-induced TNF-alpha production from macrophages was inhibited by treatment with flavone (luteolin, apigenin, and chrysin), flavonol (quercetin and myricetin), flavanonol (taxifolin), and anthocyanidin (cyanidin chloride) in vitro. Most of these, however, did not affect mice when administered orally. Serum TNF-alpha production was inhibited only by luteolin or apigenin, and only luteolin or quercetin inhibited 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced ear edema. These results suggest that the structure of luteolin: 3',4',5,7-tetrahydroxyflavone, is most suitable for the oral anti-inflammatory activity and that existence or disappearance of a hydroxy group may cause a loss of efficiency. PMID- 14745175 TI - Bacillus subtilis ypgA gene is fni, a nonessential gene encoding type 2 isopentenyl diphosphate isomerase. AB - We previously identified the fni gene of Streptomyces sp. strain CL190 as type 2 isopentenyl diphosphate (IPP) isomerase, which needs both FMN and NADPH for enzyme activity. An fni gene homolog, ypgA, was detected in the database of the Bacillus subtilis genome. However, the ypgA product was about 140 amino acids shorter in the N-terminal than the Streptomyces fni gene product. A database search found three new putative start codons in 129, 225, and 411 bases upstream of the original start codon of the ypgA gene. The longest gene product, which was named ypgA3, showed the most significant homology to the Streptomyces fni gene product. The ypgA3 gene was expressed with an N-terminal His-tag in Escherichia coli and the purified soluble protein was characterized in detail. The ypgA3 protein converted IPP to its isomer dimethylallyl diphosphate in the presence of both FMN and NADPH. The enzyme also catalyzed the reverse reaction in the presence of both the cofactors. Disruption of the ypgA3 gene was not lethal to B. subtilis. These results indicate that Bacillus ypgA3 gene is fni, a nonessential gene encoding type 2 IPP isomerase. PMID- 14745176 TI - Royal jelly inhibits the production of proinflammatory cytokines by activated macrophages. AB - In this study, we have examined the anti-inflammatory actions of royal jelly (RJ) at a cytokine level. When supernatants of RJ suspensions were added to a culture of mouse peritoneal macrophages stimulated with lipopolysaccharide and IFN-gamma, the production of proinflammatory cytokines, such as TNF-alpha, IL-6, and IL-1, was efficiently inhibited in a dose-dependent manner without having cytotoxic effects on macrophages. This suggests that RJ contains factor(s) responsible for the suppression of proinflammatory cytokine secretion. We named the factor for honeybees RJ-derived anti-inflammatory factor (HBRJ-AIF), and further investigated the molecular aspects of it. Size fractionation study showed that HBRJ-AIF is composed of substances of low (< 5 kDa) and high (> 30 kDa) molecular weights, with the former being a major component. Chromatographic analysis showed that MRJP3 is one candidate for the HBRJ-AIF with high molecular weights. Thus, our results suggest that RJ has anti-inflammatory actions through inhibiting proinflammatory cytokine production by activated macrophages. PMID- 14745178 TI - Chiral discrimination of branched-chain fatty acids by reversed-phase HPLC after labeling with a chiral fluorescent conversion reagent. AB - Anteiso fatty acids having 16 to 29 carbon atoms were labeled with the chiral fluorescent conversion reagents, (1R,2R)- and (1S,2S)-2-(2,3 anthracenedicarboximido)cyclohexanol. The diastereomeric esters of anteiso acids having up to 20 carbon atoms were separated into two peaks in an ODS column under low column-temperature conditions, while those having more than 21 carbon atoms were not separated. A C30 column made it possible to separate diastereomeric esters up to C29 anteiso acid. It was possible to predict the absolute configuration of each acid by the elution order of the derivatives. PMID- 14745177 TI - Cloning of a gene cluster responsible for the biosynthesis of diterpene aphidicolin, a specific inhibitor of DNA polymerase alpha. AB - The fungal diterpene, aphidicolin, is a well-known specific inhibitor of DNA polymerase alpha. Terpenoids are an important class of natural products. However, identification of the biosynthetic gene cluster in terpenoids is relatively rare compared with another important class of natural products, polyketides. To explore a reliable identification method for the biosynthetic gene cluster in fungal diterpenoids, cloning of the biosynthetic gene cluster of aphidicolin was employed. The application of a simple PCR method for genome walking based on the sequence of cDNA encoding aphidicolan-16beta-ol synthase (ACS) allowed us to analyze a 15.6-kb region of the Phoma betae genomic DNA. Six ORFs, PbGGS, ACS, PbP450-1, PbP450-2, PbTP, and PbTF were found in this region, and respectively expected to encode geranylgeranyl diphosphate synthase, diterpene synthase, two cytochrome P-450s, the transporter and transcription factor. Their amino acid sequences and introns were deduced by a corresponding cDNA analysis. This study shows that simple PCR-based genome walking without constructing a genomic DNA library is useful for identification of a small gene cluster. We propose a general strategy for the cloning the biosynthetic genes of fungal diterpenoids by using fungal GGS. PMID- 14745179 TI - The primary structure of a novel goose-type lysozyme from rhea egg white. AB - G-type lysozyme is a hydrolytic enzyme sharing a similar tertiary structure with plant chitinase. To discover the relation of function and structure, we analyzed the primary structure of new G-type lysozyme. The complete 185 amino acid residues of lysozyme from rhea egg white were sequenced using the peptides hydrolyzed by trypsin, V8 protease, and cyanogen bromide. Rhea lysozyme had sequence similarity to ostrich, cassowary, goose, and black swan, with 93%, 90%, 83%, and 82%, respectively. The six substituted positions were newly found at positions 3 (Asn), 9 (Ser), 43 (Arg), 114 (Ile), 127 (Met), and 129 (Arg) when compared with ostrich, cassowary, goose, and black swan lysozymes. The amino acid substitutions of rhea lysozyme at subsite B were the same as ostrich and cassowary lysozymes (Ser122 and Met123). This study was also constructed in a phylogenetic tree of G-type lysozyme that can be classified into at least three groups of this enzyme, namely, group 1; rhea, ostrich, and cassowary, group 2; goose, black swan, and chicken, and group 3; Japanese flounder. The amino acid sequences in assembled three alpha-helices found in this enzyme group (Thammasirirak, S., Torikata, T., Takami, K., Murata, K., and Araki, T., Biosci. Biotechnol. Biochem., 66, 147-156 (2002)) were also highly conserved, so that they were considered to be important for the formation of the hydrophobic core structure of the catalytic site and for maintaining a similar three-dimensional structure in this enzyme group. PMID- 14745180 TI - Metofluthrin: a potent new synthetic pyrethroid with high vapor activity against mosquitoes. AB - (1R)-trans-Norchrysanthemic acid fluorobenzyl esters are synthesized and their structure-activity relationships are discussed. These esters show outstanding insecticidal activity against mosquitoes. In particular, the 2,3,5,6-tetrafluoro 4-methoxymethylbenzyl analog (metofluthrin) exhibits the highest potency, being approximately forty times as potent as d-allethrin in a mosquito coil formulation when tested against southern house mosquitoes (Culex quinquefasciatus). Metofluthrin also exhibits a significant vapor action at room temperature. PMID- 14745181 TI - Frost-susceptible protein in plasma membranes in tubers of Helianthus tuberosus L. AB - When plasma membranes were prepared from tubers of Helianthus tuberosus L. (Jerusalem artichoke) frozen at a sublethal temperature (-10 degrees C), the levels of some plasma membrane proteins, named frost-susceptible proteins (FSPs), decreased [Uemura, M., et al., Plant Physiol., 80, 187-195 (1986)]. The aim of this study was to characterize the response of FSP120, which is named FSP-3 in a previous report, to freezing treatment by immunoblotting. Levels of FSP120 in the plasma membranes of tubers decreased after sublethal freezing, whereas no degraded products were detected in the microsomes or the soluble fraction. The amount of FSP120 in the crude extract of frozen tubers remained at a comparable level to that of the unfrozen tubers. These results suggest that FSP120 might be released from plasma membranes during freezing treatment of the tubers of Jerusalem artichoke. PMID- 14745182 TI - Synthesis and antioxidant activity of oxygenated furofuran lignans. AB - Nine furofuran compounds having a different type of oxidation were synthesized from one common intermediate in a short series of steps, and the antioxidant activity was evaluated. It was found that the tertiary hydroxy group on the furofuran ring affected the degree of antioxidant activity and that the structure, except for the phenolic part, was important for the antioxidant activity. PMID- 14745183 TI - Preparation of selenium-enriched sprouts and identification of their selenium species by high-performance liquid chromatography-inductively coupled plasma mass spectrometry. AB - Sprouts of several plants (10 families and 28 species) were cultivated in a high selenium environment, and the chemical species of selenium in these selenium enriched sprouts were identified by using high-performance liquid chromatography inductively coupled plasma mass spectrometry (HPLC-ICPMS). Cultivation of sprouts of kaiware daikon (type of radish) with 5.0 microg/ml or 10.0 microg/ml of selenium as selenite inhibited the growth. However, no abnormalities in the shape or color were apparent even in the sprouts exposed to 10.0 microg/ml of selenium. The selenium concentration in the sprouts of most plants examined was higher than that from environmental exposure. Among the types of selenium that were accumulated, a large part (69-98%) was extractable in 0.2 M HCl. Chemical analysis of selenium in the HCl extract showed that the main selenium species in all the sprouts examined was Se-methylselenocysteine. In addition to Se methylselenocysteine, selenomethionine, non-metabolized selenite, gamma-glutamyl Se-methylselenocysteine and an unknown selenium compound were also detected in several high-selenium sprouts. Since higher anticarcinogenic activities of these monomethylated selenoamino acids have been observed, it is anticipated that such selenium-enriched sprouts will be used as a foodstuff for cancer prevention. PMID- 14745184 TI - Tomato paste fraction inhibiting the formation of advanced glycation end products. AB - A water-soluble and low-molecular-weight fraction (SB) was obtained from tomato paste. The effects of SB on the formation of advanced glycation end-products (AGE) in protein glycation were studied by the methods of specific fluorescence, ELISA and a Western blot analysis, using the anti-AGE antibody after incubating protein with sugar. The results suggest that SB had strong inhibitory activity, in comparison with aminoguanidine as a positive control, and that the inhibitory mechanism of SB differed from that of aminoguanidine to involve trapping of reactive dicarbonyl intermediates in the early stage of glycation. SB contained an antioxidant, rutin, which showed potent inhibitory activity. The results also suggest that rutin chiefly contributed to inhibiting the formation of AGE, and that other compounds in SB may also have been related to the activity. PMID- 14745185 TI - Self-cloning yeast strains containing novel FAS2 mutations produce a higher amount of ethyl caproate in Japanese sake. AB - Point mutation of Gly1250Ser (1250S) of the yeast fatty acid synthase gene FAS2 confers cerulenin resistance. This mutation also results in a higher production of the apple-like flavor component ethyl caproate in Japanese sake. We mutated the 1250th codon by in vitro site-directed mutagenesis to encode Ala (1250A) or Cys (1250C) and examined cerulenin resistance and ethyl caproate production. The mutated FAS2 genes were inserted into a binary plasmid vector containing a drug resistance marker and a counter-selectable marker, GALp-GIN11M86. The plasmids were integrated into the wild-type FAS2 locus of a sake yeast strain, and the loss of the plasmid sequences from the integrants was done by growth on galactose plates, which is permissive for loss of GALp-GIN11M86. These counter-selected strains contained either the wild type or the mutated FAS2 allele but not the plasmid sequences, from which FAS2 mutant strains were selected by allele specific PCR. The FAS2-1250C mutant produced a higher amount of ethyl caproate in sake than FAS2-1250S, while FAS2-1250A produced an ethyl caproate level intermediate between FAS2-1250S and the parental Kyokai no. 7 strain. Interestingly, these mutants only showed detectable cerulenin resistance. These 'self-cloning' yeast strains should be acceptable to the public because they can improve sake quality without the presence of extraneous DNA sequences. PMID- 14745186 TI - Effects of several variable factors on the isotope ratio by HRGC-MS. AB - In the isotope ratio (Ir) analysis using GC-MS, several variable factors in sampling incidental to any food analysis were investigated for yuzu fruit. The Irs of ten monoterpene hydrocarbons in yuzu essential oils from each of six fruiting positions of three trees were measured. The sign test following t-test of all the Ir values demonstrated that there was no significant difference between both sampling years of 2001 and 2002. There was also no significant variation in the Ir values among the three trees and six fruiting positions in the individual two years. PMID- 14745187 TI - sigma marY1, the LTR of the gypsy-type retroelement marY1 from the basidiomycete tricholoma matsutake, allows multicopy DNA integration in Lentinula edodes. AB - sigma marY1 is the LTR of the retroelement marY1 from the homobasidiomycete Tricholoma matsutake. Upon integration through transformation, pLC1-hph carrying a sigma marY1 derivative, sigma* marY1, conferred the hygromycin-resistant phenotype stronger than the vector without sigma* marY1 on Lentinula edodes. Based on the densitometric analysis after Southern hybridization, a copy number of the former construct integrated in the genome is much higher than that of the latter. We conclude that sigma marY1 allows multicopy DNA integration and will be useful in the genetic research on this fungal group. PMID- 14745188 TI - Detection of hydrolytic activity of trypsin with a fluorescence-chymotryptic peptide on a TLC plate. AB - To find a new trypsin-like enzyme, a simple assay method of the hydrolysis activity for trypsin has been found. We used 6-aminoquinolyl-N hydroxysuccinimidyl carbamate (AQC) in the peptide labeling as a substrate for the trypsin-like peptidase in this study. The peptidase activity of trypsin was detected by using an AQC-chymotryptic peptide (AHP1) obtained from bovine hemoglobin. This showed that the substrate specificity of trypsin-like peptidase was distinguishable from that of the others by this procedure, and the method was used extensively in cases of various trypsin inhibitors with no significant interference from the concomitant. PMID- 14745189 TI - SMXA-5 mouse as a diabetic model susceptible to feeding a high-fat diet. AB - The SMXA-5 strain, a new mouse model for type 2 diabetes, is a recombinant inbred strain derived from non-diabetic SM/J and A/J strains. As dietary fat is a key component in the development of diabetes, we compared the glucose tolerance and diabetes-related traits among the SMXA-5, SM/J, and A/J strains while feeding a high-fat diet for 10 weeks. SMXA-5 fed on a high-fat diet showed an increased serum insulin concentration. Judging from the hyperinsulinemia in SMXA-5, this strain showed insulin resistance, an inability of peripheral tissues to respond to insulin, which was strengthened by feeding with a high-fat diet. When fed on a high-fat diet for 5 weeks, the SMXA-5 mice showed severely impaired glucose tolerance. On the other hand, SM/J showed mildly impaired glucose tolerance, even when fed on a high-fat diet for 10 weeks. These results indicate that SMXA-5 would be available for use as a diabetic model susceptible to a high-fat diet. PMID- 14745190 TI - Distribution of minerals in quinoa (Chenopodium quinoa Willd.) seeds. AB - The distribution of minerals in quinoa (Chenopodium quinoa Willd.) seed was examined using energy dispersive X-ray microanalysis (EDX) in combination with scanning electron microscopy (SEM). Phosphorus, K, and Mg coincided in localization in embryonic tissue. Since phytin globoids have been known to localize in protein bodies in embryonic cells of quinoa seed, it is thought that P is attributed to phytic acid and that K and Mg form to phytate. Calcium and K were present in the pericarp, where the cell wall is thickly developed, suggesting that these minerals are associated with pectin. Sulfur occurred in embryonic tissues, which would be derived from sulfur amino acid residues of storage proteins concentrated in the tissues. Abrasion of quinoa seeds resulted particularly in decrease in Ca content. PMID- 14745191 TI - High-level production of hyperthermophilic cellulase in the Bacillus brevis expression and secretion system. AB - A hyperthermophilic cellulase derived from Pyrococcus horikoshii was successfully produced with the Bacillus brevis host-vector system. The production of the recombinant enzyme was increased about 20-fold (to a level of 100 mg per liter) by the insertion of certain amino acid such as alanine and peptides like AEEAADP between the carboxyl end of signal peptide and the N-terminus of the mature cellulase. These recombinant cellulases had the same characteristics as that of the cellulase expressed in Escherichia coli. PMID- 14745192 TI - Suppressive effects of selected antioxidants on the activated leukocytes-induced mutagenesis in the co-culture assay systems. AB - We recently established a novel co-culture assay system using activated inflammatory cells and AS52 Chinese hamster ovary cells, and demonstrated that reactive oxygen and nitrogen species (RONS) generated from activated inflammatory leukocytes induce mutations in the gpt recorder gene in AS52 cells. In this study, we examined the inhibitory effects of 19 agents with antioxidative properties on RONS generation in cultured inflammatory cells and on mutagenesis in AS52 cells co-cultured with activated inflammatory cells. The results demonstrate that there is a linear correlation between the ability of these agents to suppress RONS production in activated inflammatory cells and to inhibit mutation in AS52 cells. PMID- 14745193 TI - High phosphorus diet changes phosphorus metabolism regardless of PTH action in rats. AB - In this study, we ascertained whether the parathyroid hormone (PTH) dominantly regulated the effects of high phosphorus (P) intakes on urinary excretion of P and bone metabolism in rats. To maintain serum PTH level equally, parathyroidectomy (PTX) and sham-operated rats were constantly exposed to rPTH(1 34) and fed both control (0.3% P) and high P (1.2% P) diet for 7 days, respectively. Urinary excretions of P and C-terminal telopeptides of type I collagen were significantly increased in both PTX and sham rats by the high P diet. These results suggest that high P diet increased urinary P excretion while promoting bone resorption regardless of PTH-dependent regulation. PMID- 14745194 TI - Protective effect of Blue-M1 against oxidative stress on COS-1 cells. AB - Blue-M1 is a blue pigment formed from xylose and glycine in the Maillard reaction. Previous work revealed that Blue-M1 scavenged hydroxyl radicals, and prevented the autoxidation of linoleic acid in vitro. We investigated the protective effect of Blue-M1 for 2,2'-azobis(2-amidino-propane)dihydrochloride (AAPH)-induced toxicity in COS-1 cells. COS-1 cells were cultured in AAPH containing DMEM medium with or without Blue-M1 at 37 degrees C for 24 h. Blue-M1 decreased the AAPH-induced toxicity in COS-1 cells, and this effect was dose dependent. Furthermore, COS-1 cells were treated with diphenyl-1-pyrenylphosphine (DPPP), as a reagent for the detection of lipid peroxide, and then were cultured in AAPH containing DMEM medium with or without Blue-M1 at 37 degrees C for 6 h. Blue-M1 prevented the AAPH-induced peroxidation of cell membrane on COS-1 cells, and this effect was also dose-dependent. These results suggest that Blue-M1 prevents the oxidative cell injury. Therefore, Blue-M1 will be an antioxidant, which protect against the oxidative stress in living systems. PMID- 14745195 TI - Maturation of fermented rice-koji miso can be monitored by an increase in fatty acid ethyl ester. AB - A mixture of steamed soybean and boiled rice with seeded Aspergillus oryzae was naturally fermented without addition of yeasts or Lactobacilli, and kept matured for 12 months at room temperature. Chemical analysis of this rice-koji miso sample for lipid changes during maturation showed that triacylglycerol was gradually decomposed into free fatty acid, with distinct formation of fatty acid ethyl ester which, six months after the start of fermentation, came to account for 35.0% of total lipid. The ester was constituted primarily with linoleic acid (ca. 50%) and oleic acid (ca. 20%), no appreciable change in this proportion being observed during maturation. Also, the proportion was unique in that this did not reflect the fatty acid composition in a mixture of the two materials. It is possible to monitor the maturation of the rice-koji miso by following up the increase with time in fatty acid ethyl ester. PMID- 14745196 TI - Introduction of DPR, an enterostatin fragment peptide, into soybean beta conglycinin alpha' subunit by site-directed mutagenesis. AB - DPR, a fragment peptide of enterostatin (VPDPR) having hypocholesterolemic activity, was introduced into the three homologous sites, EPR, DYR, and DPI, in the soybean beta-conglycinin alpha' subunit by site-directed mutagenesis. The modified beta-conglycinin was expressed in Escherichia coli and recovered in the soluble fraction. After purification on ion-exchange HPLC, the modified beta conglycinin was digested by trypsin to release integrated DPR. The yield of DPR from 1 mole of the modified beta-conglycinin was 1.2 mole. PMID- 14745197 TI - Penipratynolene, a novel nematicide from penicillium bilaiae Chalabuda. AB - New acetylenic nematicidal compound, penipratynolene (1), methy (2'R)-4-(2' hydroxy-3'-butynoxy)benzoate, together with two known compounds, 6 methoxycarbonylpicolinic acid (2) and 2,6-pyridinedicarboxylic acid (3), were isolated from the culture filtrate of Penicillium bilaiae Chalabuda. The structures of 1-3 were established by spectroscopic methods. The absolute configuration of 1 was confirmed by using a modified version of Mosher's method. Compounds 1-3 showed nematicidal activity of 77%, 52%, and 98%, respectively, by a bioassay at 300 mg/l with the root-lesion nematode Pratylenchus penetrans. PMID- 14745198 TI - A new prenylated flavonoid from propolis collected in Okinawa, Japan. AB - The new prenylflavonoid, isonymphaeol-B (1), together with three known compounds, nymphaeol-A (2), nymphaeol-B (3), and nymphaeol-C (4), were isolated from propolis collected in Okinawa, the southern-most prefecture of Japan. The structure of each compound was determined by spectral methods, including mass spectrometry and 2D NMR. Each compound had 1,1-diphenyl-2-picryl-hydrazyl radical scavenging activity. PMID- 14745199 TI - Mac1 positive cells are required for enhancement of splenocytes proliferation caused by bisphenol a. AB - We examined the effects of bisphenol A (BPA) on immune cells and it was shown that BPA upregulated the proliferation of murine splenocytes stimulated with Concanavalin A (ConA). The upregulating effects of BPA were removed with depleting Mac1+ cells from the splenocytes. This study provides evidence for the first time that Mac1+ cells were required for enhancement of splenocytes proliferation caused by bisphenol A. PMID- 14745200 TI - Truncated Sla1 induces haploid meiosis through the Pat1-Mei2 system in fission yeast. AB - We previously reported that expression of Sla1DeltaC, a truncated form of Sla1, induces ectopic meiosis in heterothallic fission yeast and this was possibly due to the inhibition of Pat1 kinase by Sla1DeltaC. Here we found mei2 mRNA and the Mei2 protein accumulated and stability of the Mei2 protein increased when Sla1DeltaC was expressed. The former two results are considered to be the consequence of de-repression of Ste11, which is the transcription factor of mei2 and negatively regulated by Pat1 kinase. The latter result reflects the consequence of deregulation of Mei2 by Pat1 kinase. In addition, Ste11 accumulated in the nucleus when Sla1DeltaC was expressed. All these data consistently support the idea that the action of Sla1DeltaC is to inactivate Pat1 kinase. PMID- 14745201 TI - Vagus nerve stimulation therapy for patients with persistent seizures after epilepsy surgery. AB - BACKGROUND: This study reports the effectiveness of vagus nerve stimulation (VNS) among patients who failed cranial surgery for intractable epilepsy. METHODS: Data were obtained from the Cyberonics VNS therapy patient outcome registry. The integrity of the systems for collecting and processing registry data was authenticated by an independent auditing agency. RESULTS: Two nonconsecutive cohorts were compared: patients who had had prior cranial surgery (CS group, n = 921) and those who had not (non-CS group, n = 3,822). For the CS group, the median reduction in seizure frequency was 42.5% after 3 months of VNS therapy, 42.9% at 6 months, 45.7% at 12 months, 52.0% at 18 months and 50.5% at 24 months. For the non-CS group, analogous rates were 47.0, 52.9, 60.0, 62.7 and 66.7%, respectively. CONCLUSION: The effectiveness of VNS is maintained during prolonged stimulation. Patients who failed prior cranial surgery did not respond quite as favorably as all other patients receiving VNS therapy. PMID- 14745202 TI - A 10-year experience with magnetic source imaging in the guidance of epilepsy surgery. AB - Magnetic source imaging (MSI) of interictal epileptiform dipoles was studied in 100 epilepsy surgery candidates. Sixty underwent surgery. MSI epileptiform data were classified as focal, regional, multifocal, scattered or none. Resections of MSI epileptiform foci were classified as extensive (EXT) versus partial or none (P/N). MSI interictal epileptiform dipoles were found in 22 of 27 anterior temporal (ATL) cases, and in 31 of 33 extratemporal (XMT) cases. Of 10 EXT ATL cases, 5 (50%) were seizure free (SF). Of 12 P/N ATL cases, 7 (58%) were SF. Of 10 nonlesional EXT XMT resections, 8 (80%) were SF. Of 10 nonlesional P/N XMT resections, 1 (10%) was SF. Neither focality of MSI data or spatial agreement of electrographic and MSI data significantly affected outcomes. PMID- 14745203 TI - Subtemporal amygdalohippocampectomy prevents verbal memory impairment in the language-dominant hemisphere. AB - BACKGROUND: In this report, we show the operative and neuropsychological results for 20 patients with medically intractable nonlesional temporal lobe epilepsy treated surgically by subtemporal amygdalohippocampectomy whose mean postoperative follow-up period was more than 6 years. METHODS: Pre- and postoperative Wechsler Adult Intelligence Scale-Revised (WAIS/-R) scores, including verbal intelligence quotient (VIQ), performance IQ (PIQ) and full-scale IQ (FIQ) scores, were determined in the 19 adults. The revised Wechsler Intelligent Scale for Children was used in a 9-year-old boy. The verbal paired associates learning test was performed pre- and postoperatively in 15 patients. These data were compared pre- and postoperatively in 10 patients in whom the language-dominant hemisphere side was operated on, and in 5 patients in whom the language-non-dominant side was operated on. RESULTS: Seizure control was achieved in 80% of patients, with improvement into Engel categories I and II without definite permanent complications except for postoperative memory impairment in one patient. VIQ improved after 2 years, and PIQ and FIQ improved after both 2 months and 2 years postoperatively. CONCLUSION: There was no significant decline in the postoperative verbal memory scores in those patients whose medial temporal structure of the language-dominant side had been removed. PMID- 14745204 TI - Peri-insular hemispherotomy: potential pitfalls and avoidance of complications. AB - Techniques for cerebral hemispherectomy have progressively evolved towards more disconnection and less excision over the last 50 years. Peri-insular hemispherotomy (PIH), as described by the senior author, has the maximal ratio of disconnection to excision among all procedures for hemispheric epilepsy. In this study, we focus on surgical complications and intraoperative anatomical observations during PIH over the last 10 years. Based on this experience, the procedure has undergone some modifications, which we detail herein. PMID- 14745205 TI - Hemorrhagic complications of microelectrode-guided deep brain stimulation. AB - BACKGROUND: The incidence of intracranial hemorrhage occurring during microelectrode-guided implantation of deep brain stimulators (DBS) for movement disorders has not been well defined. We report the incidence of hemorrhage in a large series of DBS implants into the subthalamic nucleus (STN), thalamus (VIM) and internal globus pallidus (GPi). METHODS: All DBS procedures performed by a single surgeon (P.A.S.) between June 1998 and April 2003 were included in this study. Patients had postoperative imaging (MRI or CT) 4-24 h following surgery, and all hematomas >0.2 cm(3) in volume were noted and scored as symptomatic (associated with any new neurologic deficit lasting >24 h) or asymptomatic. RESULTS: The total number of lead implants was 357. There were 5 symptomatic hematomas and 6 asymptomatic hematomas. The relative risk of hematoma (any type) per lead implant was 3.1%. The incidence of hematoma by target site was 2.5% per lead for STN-DBS, 6.7% for GPi-DBS and 0% for VIM-DBS. CONCLUSION: The overall risk of intraoperative or early postoperative symptomatic hemorrhage with microelectrode-guided DBS, over all targets, was 1.4% per lead implant. The brain target had a significant effect on the risk of hemorrhage. PMID- 14745206 TI - Effect of high-frequency stimulation of the subthalamic nucleus on subthalamic neurons: an intracellular study. AB - BACKGROUND/AIMS: The precise mechanism of action of deep brain stimulation in the subthalamic nucleus (STN) for the treatment of Parkinson's disease and epilepsy is unknown. In the present study, the intracellular effects on STN neurons following high-frequency stimulation (HFS) of STN were examined to test the hypothesis that HFS results in either an increase or a decrease in neuronal action potential generation. METHODS: Intracellular electrophysiological recordings were made in the rat STN neurons in in vitro slice preparations. A concentric bipolar stimulating electrode was placed in the STN, and electrical stimulation (duration, 100-2000 ms; amplitude, 10-500 microA, and frequency, 10 200 Hz) was delivered while simultaneously recording intracellularly from a STN neuron using a sharp electrode. RESULTS: HFS of STN resulted in the generation of excitatory postsynaptic potentials and an increase in action potential firing during the stimulation period followed by a period of poststimulation inhibition of firing in STN neurons. The degree of increase in action potentials from HFS was critically dependent on the frequency of electrical stimulation, i.e. at approximately 100-140 Hz, maximal increase was obtained, but at 200 Hz, the activity was blocked. Interestingly, the duration of poststimulation inhibition of firing was dependent on the duration of stimulation, i.e. the longer the HFS, the longer the inhibition. CONCLUSIONS: These results suggest that the mechanism of action of deep brain stimulation involves initial excitation followed by later inhibition of STN neurons at a cellular level rather than primary inhibition, as previously hypothesized. PMID- 14745207 TI - Subthalamic nucleus stimulation in Parkinson's disease: correlation of active electrode contacts with intraoperative microrecordings. AB - BACKGROUND/AIMS: The most effective site for subthalamic nucleus (STN) stimulation has remained unclear. The position of active contacts relative to the dorsal margin of the STN was determined. METHODS: Electrodes (n = 49) were implanted following STN mapping by microrecording and microstimulation along five tracks (n = 25 patients). The stereotactic position of active contacts was determined and correlated with microrecordings using an algorithm for direct three-dimensional comparisons (n = 37). RESULTS: Most active contacts were detected within +/-1.0 mm from the dorsal margin of the STN as defined by microrecording (32.4%) or farther dorsal in the subthalamic area (37.8%), and only 29.7% were localized to the STN proper. This was consistent with the average stereotactic coordinates of the active contacts in these three groups. CONCLUSION: Our data suggest that the dorsal border area of the STN is the most effective target. Besides the dorsolateral STN (sensorimotor part) this may include projections from/to STN, the zona incerta, and pallidofugal projections in the fields of Forel. PMID- 14745208 TI - Effect of motor improvement on quality of life following subthalamic stimulation is mediated by changes in depressive symptomatology. AB - BACKGROUND/AIMS: Subthalamic deep brain stimulation (STN-DBS) for Parkinson's disease (PD) improves motor symptoms and quality of life (QOL). Because depression is a potent correlate of QOL, and STN-DBS may be associated with changes in mood, this study sought to determine whether QOL improvement is a direct or indirect consequence of motor improvement. METHODS: 26 patients with PD, free of dementia and major depression, who consecutively underwent bilateral, microelectrode-guided STN-DBS, underwent preoperative and 3-month postoperative neuropsychological evaluation, including measures of QOL (PD Questionnaire -39) and depressive symptoms (Beck Depression Inventory). RESULTS: Motor score in the Unified Parkinson's Disease Rating Scale (UPDRS Part III) improved significantly with STN-DBS relative to preoperative 'on' and 'off' scores, as did QOL and depressive symptoms. Extent of QOL improvement tended to be associated with improvement in motor score from presurgical on to postsurgical on stimulation and on medication state. QOL improvement was significantly related to amelioration of depressive symptoms. Partial correlations revealed that the association between QOL improvement and depression remained significant when influence of motor improvement on QOL and depression was controlled for. The motor-QOL association was no longer significant when effects of depression were controlled for. CONCLUSIONS: Significant QOL improvements after STN-DBS are associated with improved motor 'on' state and depressive symptoms. The influence of motor improvement on QOL may be largely indirect by reducing depression. PMID- 14745209 TI - Thalamic stimulation in patients with multiple sclerosis: long-term follow-up. AB - BACKGROUND: We assessed the long-term effect of deep brain stimulation (DBS) in patients with multiple sclerosis (MS). METHODS: Nine patients with MS-induced tremor underwent placement of a DBS electrode in the thalamus. All patients were referred from the tertiary MS center at the New Jersey Medical School. Intraoperative macrostimulation was performed under local anesthesia before permanent implantation. Preoperative and postoperative evaluation included magnetic resonance imaging (MRI), the Extended Disability Status Scale (EDSS), the Bain-Finchley tremor scale, neuropsychological testing and patient assessment of the benefit from surgery. RESULTS: There were no surgical complications. Follow-up ranged from 9 to 48 months (mean 32 months). EDSS scores averaged 7.2 before surgery, 6.8 at 6 months after surgery and 7.8 at late follow-up. Tremor scores averaged 5.4 before surgery, 1.7 at 6 months after surgery and 2.1 at late follow-up. Four patients who had significant tremor reduction at 6 months had severe MS progression afterwards, and one was lost to late follow-up. Another patient, in whom excellent tremor control was obtained, developed increasing stimulation-induced fatigue, and the implant was removed at his request. The 3 other patients have maintained a worthwhile benefit from DBS. MRI scans did not show any new MS plaques in relation to the electrode, although imaging was consistent with disease progression in all patients. Neuropsychological testing showed a mild to moderate decline in cognitive function consistent with disease progression. CONCLUSIONS: Chronic thalamic stimulation decreases tremor in patients with MS. However, this improvement does not correlate with improvement in objective measures of function. Any actual benefit may be limited in most patients by the progression of underlying disease. DBS for relief of tremor in patients with MS should be considered only in carefully selected patients with relatively stable disease, in whom upper extremity tremor is a disabling symptom. PMID- 14745210 TI - Longevity of batteries in internal pulse generators used for deep brain stimulation. AB - The longevity of batteries in internal pulse generators (IPGs) used clinically for deep brain stimulation is not known. We conducted a study to assess the life span of these batteries. From 1993 to 2000, 163 single-channel batteries were surgically implanted in our institution. The electrical settings utilized in patients who had battery failures were assessed and the total electrical energy delivered (TEED) was calculated and correlated with battery longevity. Fourteen IPGs had battery failure requiring replacement. The median life span of the batteries was 45 months. Batteries with high energy consumption as assessed by TEED had a reduced life span (r = -0.82, p < or = 0.001). Patients with essential tremor who required battery replacement needed higher settings to control their symptoms and therefore presented a shorter battery life when compared to patients with Parkinson's disease. In our series of patients who needed battery replacement, battery longevity varied with stimulation parameters but was longer than expected from the manufacturer's specifications. PMID- 14745211 TI - History repeats itself. AB - Although many advances in stereotactic surgery appear to be of recent origin, there are precedents in the literature documenting the planting of those seeds that eventually grew into common procedures. Pallidotomy was the first stereotactic procedure in humans that used the Spiegel-Wycis apparatus in 1947. Other targets for Parkinson's disease have their roots in the decade following that. Using stereotactic techniques to target solid or cystic lesions was introduced in the 1950's, when either calcification or pneumoencephalography were necessary to visualize the mass. One of the first uses of computers in surgery was in stereotaxis. Infusion of a neurotransmitter was first done over 30 years ago. PMID- 14745212 TI - Subthalamic nucleus deep brain stimulation for Parkinson's disease: magnetic resonance imaging targeting using visible anatomical landmarks. AB - BACKGROUND: The aim of the present study was to validate a magnetic resonance imaging (MRI) visual procedure to target the subthalamic nucleus (STN) based on surrounding anatomical landmarks. METHODS: 31 consecutive bilaterally implanted parkinsonian patients were included in this study. After identification of the anterior commissure (AC), posterior commissure (PC) and midcommissural point on a three-dimensional T1-weighted sequence, inversion recovery (IR) T2-weighted coronal slices were performed orthogonal to the AC-PC line. On the slice showing the anterior pole of the red nucleus (RN), the target was placed in the inferolateral portion of the subthalamic zone, limited superiorly by the thalamus, laterally by the internal capsule, inferiorly by the substantia nigra and medially by the midline. The distribution of the targets was analyzed in the AC-PC referential. RESULTS: The mean target coordinates were as follows: anteroposterior (AP) = -2.54 mm (+/-1.37 mm), lateral (LAT) = 12.03 mm (+/-0.91 mm) and vertical (VERT) = -6.10 mm (+/-1.52 mm) for the right side, and AP = 2.65 mm (+/-1.36 mm), LAT = -11.97 mm (+/-1.30 mm) and VERT = -5.89 mm (+/-1.52 mm) for the left side. They projected in the inferior portion of the STN on the Schaltenbrand and Wahren atlas [Stuttgart, Thieme, 1977]. CONCLUSION: Identification of the anterior pole of the RN and the subthalamic zone on coronal IR T2-weighted MRI performed orthogonal to the AC-PC line provides a precise visual procedure to target the STN. PMID- 14745213 TI - Variability in position of the subthalamic nucleus targeted by magnetic resonance imaging and microelectrode recordings as compared to atlas co-ordinates. AB - BACKGROUND: Traditional methods for localisation of target nuclei for deep brain stimulation (DBS) have used brain atlas co-ordinates for initial targeting. It is now possible to visualise the subthalamic nucleus (STN) on magnetic resonance imaging (MRI) and determine the individual variability of its position. METHODS: The present study was performed in patients undergoing STN DBS for Parkinson's disease. The STN was directly targeted from axially obtained MRI and verified with microelectrode recordings. Postoperatively, the most effective contact was identified for each patient, and its position was calculated. RESULTS: Fifty electrodes were inserted in 25 patients. The target position varied considerably in relation to the mid-commissural point. The mean effective contact position lies just dorsal to the location of the STN in a standard brain atlas. CONCLUSION: The STN varies in position, and can be accurately targeted from MRI alone. PMID- 14745214 TI - Stereotactic ventrooralis thalamotomy for task-specific focal hand dystonia (writer's cramp). AB - BACKGROUND: Writer's cramp is a type of focal dystonia due to dysfunction of the pallido-thalamo-cortical circuit. The symptom is refractory to most conservative treatment, though botulinum toxin injection is generally used for symptomatic relief. As a surgical treatment of dystonia, we performed stereotactic nucleus ventrooralis (Vo) thalamotomy for dystonic cramp of the hand. METHODS: Twelve patients (5 men, 3 women; age 26-40 years, mean 32.1 years) with medically intractable task-specific focal dystonia of the hand underwent Vo thalamotomy. The stereotactic target was chosen at the junction of the anterior and posterior Vo nuclei. RESULTS: The mean duration of the symptom ranged from 3 to 6 years (mean 4.5 years.) All patients had complained of difficulty in writing. Seven patients were professionals, such as a comic artist, guitarist and barber, and, because of the dystonic symptoms occurring during their work, they had stopped pursuing their profession. All patients showed immediate postoperative disappearance of dystonic symptoms, and the effect was sustained during the follow-up period (3-33 months, mean 13.1 months), except in one case. Two patients showed partial recurrence of the symptom and underwent second thalamotomy 5 months after the initial surgery with satisfactory results. The score on the writer's cramp rating scale decreased significantly (p < 0.001) after Vo thalamotomy. There were no permanent operative complications. There was no mortality or permanent morbidity. CONCLUSION: Although a longer follow-up is needed, stereotactic Vo thalamotomy is a useful and safe therapeutic option for writer's cramp. PMID- 14745215 TI - A novel denervation procedure for idiopathic cervical dystonia. AB - BACKGROUND: For idiopathic cervical dystonia, the treatment of choice is botulinum toxin injection or surgical denervation. There are two major procedures of surgical denervation: intradural ventral rhizotomy and extradural peripheral neurotomy (Bertrand procedure). The Bertrand procedure is always accompanied by popstoperative sensory loss in the C2 region. METHODS: The authors have modified these procedures to minimize the complications. Our method is unilateral intradural ventral rhizotomy of C1 and C2, extradural denervation of the C3-C6 posterior rami and contralateral peripheral sectioning of the branches of the accessory nerve to the sternocleidomastoid muscle. Forty-four patients underwent this modified operation ('Taira's' method: group A) and the results were compared with those in a matched control group of 38 patients who underwent the traditional Bertrand's procedure (Bertrand's method: group B). RESULTS: Three patients in group A showed a sensory deficit in the C2 area, while all of the patients in group B had C2 sensory disturbance. Pre- and postoperative rating scores did not differ between the two groups. The intraoperative blood loss was significantly smaller in group A. CONCLUSION: Compared with the traditional Bertrand's operation, our procedure carries a much lower incidence of complications and a significant decrease of intraoperative blood loss. PMID- 14745217 TI - Effect of stereotactic high-frequency stimulation in the anterior lobe of the cerebellum in cerebral palsy: a new suboccipital approach. AB - The direct stereotactic suboccipital approach to the anterior lobe of the cerebellum was applied for deep high-frequency stimulation in three patients (for 29, 8 and 3 months) suffering from cerebral palsy. In agreement with our previous experience with a transtentorial approach in 30 patients, spasticity, dyskinesias and behavior were improved during chronic intermittent stimulation (frequency 185 Hz, pulse width 210 micros, voltage individually altered according to motor response at 0.5-4.0 V, 15 min on, 2-6 h off). Patients attained useful motor skill improvements. The results indicate that the method is safe, effective and reasonable. PMID- 14745216 TI - A quick and universal method for stereotactic visualization of the subthalamic nucleus before and after implantation of deep brain stimulation electrodes. AB - For deep brain stimulation (DBS) of the subthalamic nucleus (STN), it would be an advantage if the STN could be visualized with fast acquisition of MR images, allowing direct and individual targeting. We present a protocol for T2-weighted, nonvolumetric fast-acquisition MRI, implemented at 8 centers in 6 countries. Acquisition time varied between 3 min 5 s and 7 min 48 s according to the center, and imaging often provided visualization of the STN on axial and coronal scans. Postoperatively, the same imaging protocol permitted visualization of the target area and DBS electrodes with minimum artifacts. This imaging technique may contribute to a decrease in the number of electrode passes at surgery. PMID- 14745218 TI - Deep brain stimulation for parkinson's disease: correlation between intraoperative subthalamic nucleus neurophysiology and most effective contacts. AB - Though intraoperative neurophysiology is essential to precisely define the definitive target, little is known regarding its predictive value in defining the most effective contact for chronic deep brain stimulation. In this retrospective study, we reviewed the correlation between intraoperative neurophysiology and contacts selected for chronic stimulation. Twenty consecutive patients implanted for subthalamic nucleus (STN) stimulation were reviewed. There was no significant correlation between the electrophysiologically defined STN and the most effective contact for chronic stimulation at 3 months or at 6 months. Furthermore, there was a discrepancy between the most effective contact for rigidity versus akinesia or tremor at 3 months. Interestingly, at 3 months, the same electrode contact was maximally efficient for rigidity, akinesia and tremor in only 13 of the 39 cases. This lack of correlation did not affect the global improvement. PMID- 14745219 TI - The NASA Smart Probe Project for real-time multiple microsensor tissue recognition. AB - BACKGROUND: Remote surgery requires automated sensors, effectors and sensor effector communication. The NASA Smart Probe Project has focused on the sensor aspect. METHODS: The NASA Smart Probe uses neural networks and data from multiple microsensors for a unique tissue signature in real time. Animal and human trials use several probe configurations: (1) 8-microsensor probe (2.5 mm in diameter) for rodent studies (normal and subcutaneous mammary tumor tissues), and (2) 21 gauge needle probe with 3 spectroscopic fibers and an impedance microelectrode for breast cancer diagnosis in humans. Multisensor data are collected in real time (update 100 times/s) using PCs. RESULTS: Human data (collected by NASA licensee BioLuminate) from 15 women undergoing breast biopsy distinguished normal tissue from both benign tumors and breast carcinoma. Tumor margins and necrosis are rapidly detected. CONCLUSION: Real-time tissue identification is achievable. Potential applications, including probes incorporating nanoelectrode arrays, are presented. PMID- 14745220 TI - A noncontacting 3-D digitizer for use in image-guided neurosurgery. AB - Current neuronavigational systems face two primary challenges: (1) automatic and robust registration between preoperative images and the operating room space, and (2) compensation for brain deformations that compromise the accuracy of the initial registration. To contend with these difficulties, we firstly estimate the three-dimensional (3-D) structure of the cortical surface using a noncontacting 3 D digitizer. This 3-D structure is then used to establish the initial registration, and to update the preoperative MR volume as the brain deforms. We show that this approach improves the accuracy of registration in a phantom study, and demonstrate the ability to capture cortical motion in six clinical cases. PMID- 14745221 TI - Functional magnetic resonance imaging in a low-field intraoperative scanner. AB - BACKGROUND: Functional magnetic resonance imaging (fMRI) has been used for preoperative planning and intraoperative surgical navigation. However, most experience to date has been with preoperative images acquired on high-field echoplanar MRI units. We explored the feasibility of acquiring fMRI of the motor cortex with a dedicated low-field intraoperative MRI (iMRI). METHODS: Five healthy volunteers were scanned with the 0.12-tesla PoleStar N-10 iMRI (Odin Medical Technologies, Israel). A finger-tapping motor paradigm was performed with sequential scans, acquired alternately at rest and during activity. In addition, scans were obtained during breath holding alternating with normal breathing. The same paradigms were repeated using a 3-tesla MRI (Siemens Corp., Allandale, N.J., USA). Statistical analysis was performed offline using cross-correlation and cluster techniques. Data were resampled using the 'jackknife' process. The location, number of activated voxels and degrees of statistical significance between the two scanners were compared. RESULTS: With both the 0.12- and 3-tesla imagers, motor cortex activation was seen in all subjects to a significance of p < 0.02 or greater. No clustered pixels were seen outside the sensorimotor cortex. The resampled correlation coefficients were normally distributed, with a mean of 0.56 for both the 0.12- and 3-tesla scanners (standard deviations 0.11 and 0.08, respectively). The breath holding paradigm confirmed that the expected diffuse activation was seen on 0.12- and 3-tesla scans. CONCLUSIONS: Accurate fMRI with a low-field iMRI is feasible. Such data could be acquired immediately before or even during surgery. This would increase the utility of iMRI and allow for updated intraoperative functional imaging, free of the limitations of brain shift. PMID- 14745222 TI - Use of the NeuroMate stereotactic robot in a frameless mode for movement disorder surgery. AB - BACKGROUND/AIMS: To evaluate the use of the NeuroMate stereotactic robot with a novel ultrasound registration system for movement disorder surgery (MDS). METHODS: Using the robot in a frameless mode, 51 patients underwent MDS. Surgical planning was carried out using MRI data obtained more than 24 h before surgery. RESULTS: 37 out of 50 targets in the subthalamic nucleus were satisfactorily identified with a single microelectrode trajectory and the final electrode positions were at a mean distance of 1.7 mm from the calculated target. There was a significant improvement in motor scores of the Unified Parkinson's Disease Rating Scale III (off medication) at 6 (43%) and 18 months (51.7%) compared to pre-operative scores (p < 0.05). CONCLUSIONS: The frameless robot using only MRI data can be used for MDS. The temporal separation of imaging from the surgical procedure provides additional time for detailed image analysis and planning. PMID- 14745223 TI - Integration of neurosurgical image guidance and an intraoperative magnetic resonance scanner. The University Hospitals of Cleveland experience. AB - BACKGROUND: Intraoperative magnetic resonance (MR) imaging has been employed as an alternative to image guidance using preoperative images. We integrated both systems to evaluate their clinical use. METHODS: The BrainLAB VectorVision system was integrated in an intraoperative Siemens Open Viva 0.2-tesla MR system. Clinical experience was assessed. RESULTS: Patterns of intraoperative imaging emerged, and benefit was seen in registering preoperative and intraoperative images. CONCLUSIONS: This integrated system has clinically observed effects on imaging, navigation, and surgery. PMID- 14745224 TI - Stereotactic accuracy of a 3-tesla magnetic resonance unit. AB - Recently, magnetic resonance imagers (MRIs) with 3-tesla magnets were approved for clinical use. The spatial accuracy of these high-resolution scanners has yet to be proven. In the present study, a computed tomography (CT)- and MRI compatible phantom was scanned on a CT, a 1.5-tesla MRI and a 3-tesla MRI scanner. The model was registered to the images using an infrared-based surgical navigation system. The distance between the predicted position of the navigation probe tip and the actual target on the image was measured on the x, y and z axes for 13 points on each image. Error was compared across imaging modalities, peripheral versus central targets and along each axis. We found that 3-tesla MRI scans are accurate as stereotactic data sets. PMID- 14745225 TI - Two-dimensional IgE-binding spectrum of Japanese cedar (Cryptomeria japonica) pollen allergens. AB - BACKGROUND: Japanese cedar (Cryptomeria japonica) pollen is one of the most prevalent sources of the allergens that elicit rhinitis and conjunctivitis. Only Cry j 1 and Cry j 2 have been well characterized as the major allergens of this pollen. OBJECTIVES: The aims of this study were to complete the repertoire of C. japonica pollen allergens, to investigate their variability with respect to IgE reactive patterns and to identify the isoforms of Cry j 1 and Cry j 2 by proteome analysis. METHODS: Proteins in C. japonica pollen separated on two-dimensional (2 D) polyacrylamide gel electrophoresis were immunodetected with IgE in sera of 40 subjects allergic to C. japonica pollen. Mass fingerprinting was used to elucidate the diversity of the major allergens. RESULTS: 2-D immunolabeling with individual patients' sera showed the distinguishable IgE-binding patterns inlaid with 4-87 spots from a total of 131 IgE-binding protein spots. At least 12 Cry j 1 (27.5-75% of IgE-binding frequency) and 3 Cry j 2 (32.5-40%) isoforms were localized. In total, 31 spots were found to be more reactive than the highest IgE reactive isoform of Cry j 2. CONCLUSIONS: The proteomics approaches showed great interindividual variation of IgE-binding patterns to C. japonica proteins and contributed to the repertoire of numerous C. japonica allergens other than Cry j 1 and Cry j 2. Protein microsequencing demonstrated more complicated multiplicity in Cry j 1 than previously known and new isoforms in Cry j 2. PMID- 14745226 TI - Partial amino acid sequence of a cellulase-like component with IgE-binding properties from Stachybotrys chartarum. AB - BACKGROUND: The aim of this study was to characterize the amino acid sequence of a selected Stachybotrys chartarum component and to investigate human IgE reactivity against components of S. chartarum and nine other fungal species. METHODS: Human IgE reactivity against S. chartarum and nine other fungal extracts was investigated by the immunoblotting method. For automated amino acid sequencing analyses, the S. chartarum extract was purified by ion exchange chromatography prior to in-gel alkylation and digestion with modified trypsin. RESULTS: Human IgE reactivity was detected against eight components in the S. chartarum extract. Over 80% of the sera from the exposed subjects and less than 50% of the control sera recognized the 33-, 48- and 50-kD S. chartarum components. The human sera detected a 48- to 50-kD component from the extracts of eight fungal species. Nineteen peptide sequences were identified from the 48-kD component of S. chartarum. An analysis of the peptide sequences revealed homology with known fungal glycoside hydrolase enzymes (cellulases). CONCLUSIONS: The data showed human IgE reactivity against several S. chartarum components, including one at 48 kD. On the other hand, the human sera recognized 48- to 50-kD components from seven other fungal species, suggesting shared antigenic components (e.g. enolase) between the fungi. Thus, to our knowledge, this is the first antigen identified from S. chartarum. PMID- 14745227 TI - Antigen-specific versus total immunoglobulin synthesis: total IgE and IgG1, but not IgG2a levels predict murine antigen-specific responses. AB - BACKGROUND: Induction of an effective antibody (Ab) response requires delivery of multiple signals to B cells. Cross-linking of the B cell antigen receptor (BCR), signaling through CD40 and CD80/86 and cytokine signals combine to induce class switching and expression of specific isotypes. These signals are principally derived from activated, antigen (Ag)-specific T cells. In contrast, IFNgamma, the only cytokine known to induce class switch to IgG2a, can be produced systemically by activated NK or NKT cells, suggesting that Ag-nonspecific signals may also regulate IgG2a production. METHODS: Given the potential differences in regulation between IgE/IgG1 versus IgG2a, we immunized mice on day 0 with ovalbumin (OVA) in the presence of strong type-1- or type-2-immunity-inducing adjuvants and boosted mice 4 weeks later. Mice were bled during the primary immune response and after boost to assess primary and recall Ab responses. RESULTS: Regardless of strain of mice used, phenotype (type 1 versus type 2 dominated) or nature of the immune response induced (primary versus recall), strong correlations between OVA specific and total IgE and IgG1 were demonstrated. In contrast, a consistent lack of correlation between OVA-specific and total IgG2a levels was observed in all but BALB/c mice. CONCLUSION: These data indicate that the increase in total levels of IgE/IgG1 isotypes is primarily a result of increased levels of OVA specific Ab. In contrast, the lack of correlation between total and OVA-specific IgG2a suggests broader activation of IgG2a-producing B cells routinely occurs following exogenous Ag immunization. PMID- 14745229 TI - A critical review of the use and application of the baboon as a model for research in women's reproductive health. PMID- 14745228 TI - Induction of TNF-alpha autoantibody production by AutoVac TNF106: a novel therapeutic approach for the treatment of allergic diseases. AB - BACKGROUND: Cytokines play an integral role in the coordination and persistence of allergic inflammatory processes and therefore represent prime targets for novel therapies in diseases such as asthma. Multiple attempts to generate low molecular-weight cytokine inhibitors have failed, and the main attention has focused on the development of biological agents such as neutralizing antibodies. The present work describes a simple and effective method to induce the production of therapeutic anti-cytokine autoantibodies by active immunization against a modified endogenous cytokine. METHODS: Balb/c mice were subcutaneously injected with AutoVac TNF106, a recombinant murine TNF-alpha molecule containing a foreign immunogenic T helper epitope, and the induction of neutralizing anti-TNF-alpha autoantibodies was analysed. These mice were then sensitized with ovalbumin (OVA), and the effect of neutralizing anti-TNF-alpha autoantibodies on the allergen-induced airway inflammation was analysed. RESULTS: AutoVac TNF106 immunized mice developed high titres of neutralizing anti-TNF-alpha autoantibodies, which were maintained for at least 4 weeks after the last booster injection. Mice vaccinated with AutoVac TNF106 and further immunized against OVA showed diminished TNF-alpha levels in the bronchoalveolar lavage (BAL) fluid after OVA challenge. Moreover, pretreatment with AutoVac TNF106 resulted in significantly reduced numbers of eosinophils and neutrophils in BAL fluid in response to single or multiple allergen exposure. CONCLUSION: The induction of anti-TNF-alpha autoantibody production by the AutoVac TNF106 technology not only confirmed the role of TNF-alpha in the induction of allergic inflammation but also offers a novel approach to block the activity of cytokines in order to treat allergic inflammatory conditions. PMID- 14745230 TI - NOPPAIN: a nursing assistant-administered pain assessment instrument for use in dementia. AB - The Non-Communicative Patient's Pain Assessment Instrument (NOPPAIN) is a nursing assistant-administered instrument for assessing pain behaviors in patients with dementia. This study investigated the validity of the NOPPAIN. Twenty-one nursing assistants (NAs) with no prior training in using the NOPPAIN watched six videos, each portraying a bed-bound patient with severe dementia receiving personal care from a nursing assistant and responding with a different level of pain intensity. The NAs completed a NOPPAIN rating for each video. The NAs were also presented with each possible pair of videos and asked to identify the video showing the most pain. Results indicated the NAs were quite accurate in their ratings of the videos, providing excellent preliminary evidence on the use of the NOPPAIN for detecting pain in nursing home patients with dementia. PMID- 14745231 TI - Overexpression of TRAP in the enamel matrix does not alter the enamel structural hierarchy. AB - The secreted, full-length amelogenin is the dominant protein of the forming enamel organ. As enamel mineralization progresses, amelogenin is quickly subjected to proteolytic activity, and eliminated from the enamel environment. Mature enamel contains only traces of structural proteins, including enamelin and the sheath protein ameloblastin. In addition, a proteolytic fragment of amelogenin, known as the tyrosine-rich amelogenin peptide or TRAP, is present in low but isolatable quantities. By overexpressing TRAP during enamel development we sought to determine if such overexpression would result in structural alterations to the mature enamel. We reasoned that overexpressing a protein associated with enamel maturation, at an inappropriate developmental stage, would result in alterations to the enamel protein assembly and hence, alterations in enamel structure and morphology. As judged by transmission and scanning electron microscopy, the enamel formed by overexpressing TRAP showed little morphological differences when compared to the enamel of normal nontransgenic animals. Based on scanning electron-microscopic images, there was modest hypomineralization evident in the interrod enamel of the TRAP-overexpressing animals. However, this finding was inconsistent and inconsequential from a structural and functional perspective. From these results it appears that additional amounts of TRAP protein in the immature enamel matrix are not sufficient to alter the properties of the enamel extracellular matrix to an extent that the hierarchical structure of mature enamel is altered. PMID- 14745232 TI - Immunolocalization of Alk8 during replacement tooth development in zebrafish. AB - The novel type I transforming growth factor-beta (TGF-beta) family member receptor Alk8 was previously identified in a degenerate RT-PCR screen for zebrafish type I and II TGF-beta family member receptors. Functional analyses revealed that Alk8 acts through Bmp signaling pathways in early embryonic dorsoventral patterning, in neural crest cell specification, and in patterning and differentiation of neural crest cell-derived pharyngeal arch cartilages. In addition, Alk8 forms active signaling complexes with TGF-beta1 and the TGF-beta RII receptor, suggesting that Alk8 mediates cross talk between Bmp and TGF-beta subfamily members. In this study, immunohistochemical analysis was performed on zebrafish aged 2 days postfertilization to 1 year, revealing immunolocalization of Alk8 to tissues of the tooth-bearing ceratobranchial 5 (cb5) arch including dental epithelial and mesenchymal tooth tissues of developing primary and replacement teeth, mucous-producing crypt epithelium, keratinized bite plate, and developing taste buds. These results suggest roles for Alk8 in patterning tooth bearing pharyngeal epithelium, in the initiation of tooth development, in odontoblast and ameloblast differentiation, and in osteoblast maturation. The ability for zebrafish to continuously form teeth throughout their lives allows for the comparison of Alk8 expression in both primary and replacement tooth development, revealing identical Alk8 expression profiles. This study advances our current understanding of the functions of Alk8, particularly with respect to primary and replacement tooth formation, reveals additional roles for Alk8 in dental epithelial patterning and in odontoblast, ameloblast and osteoblast differentiation, and demonstrates the utility of the zebrafish as a model for primary and replacement tooth development. PMID- 14745233 TI - Identification of potential modifiers of Runx2/Cbfa1 activity in C2C12 cells in response to bone morphogenetic protein-7. AB - Treatment with BMP-7 causes a shift in the differentiation pathway from myoblastic to osteoblastic in C2C12 mouse myoblast precursor cells in vitro. The underlying molecular mechanism is largely unknown. BMP-7 at 200 ng/ml completely inhibited myotube formation in C2C12 cells and dramatically induced alkaline phosphatase activity up to 20-fold when compared to untreated cells by day 12 in culture. The level of Runx2/Cbfa1 mRNA, a bone-specific transcription factor, was also stimulated up to 6-fold by BMP-7 with a peak at 24 h. In addition BMP-7 treatment stimulated a 55-fold increase in osteocalcin mRNA as early as 24 h after treatment. A novel finding was that the expression of the chondrocyte markers Sox9 and type II collagen was increased as well. Runx2/Cbfa1 is a molecular switch for osteoblast differentiation. To initiate the study of modulators of Runx2/Cbfa1, such as kinases and cofactors, during osteoblastic differentiation of C2C12 cells treated by BMP-7 in vitro, microarray analyses of gene expressions were performed. Microarray data suggested that a total of 882 transcripts were either up- or downregulated at least 2-fold. Cluster analyses revealed 76 genes (including ESTs) with expression patterns that paralleled Runx2/Cbfa1. Thirteen of these 76 genes were initially selected as potential transcription modulators for further study; including CCAAT/enhancer binding protein delta, distal- less homeobox 1, forkhead box F2, insulin-like growth factor binding protein 4, an ortholog of human osteoclast stimulating factor 1 and p300/CBP-associated factor. Some transcription modulators have been associated with osteoblastic differentiation or interacted with Runx2/Cbfa1. Most of them have not been extensively studied in osteoblastic differentiation and in relationship to Runx2/Cbfa1. Thus, these studies identify potential regulators for Runx2/Cbfa1 and osteoblast differentiation. In addition, our data revealed for the first time that BMP-7 not only induced the expression of osteoblastic differentiation markers but also stimulated the expression of chondroblastic markers in C2C12 cells. PMID- 14745234 TI - New chondrocyte genes discovered by representational difference analysis of chondroinduced human fibroblasts. AB - This report includes a review of the potential for gene expression analyses to provide new information for solving problems in skeletal repair and regeneration. It focuses on two approaches: high-throughput gene array methods and representational difference analysis (RDA). The principles underlying these methods are presented with experimental tutorials and some applications. Second, this report includes a review of results from applying both approaches to an in vitro model of postnatal chondroinduction by demineralized bone powder (DBP). Human dermal fibroblasts (hDFs) cultured with DBP acquire a chondroblast phenotype and express cartilage-specific matrix proteins after 7 days. We used cDNA macroarrays and RDA to identify the genes that were altered prior to expression of the chondroblast phenotype, i.e., after only 3 days' culture with DBP. Using a strategy of data management and reduction based upon biological functions, we reported several functional families of genes (cytoskeletal elements, protein synthesis/trafficking, and matrix molecules and their modifiers) that are upregulated during chondroinduction of hDFs. Together with histological and biochemical evidence of the chondroblast phenotype, the gene expression patterns indicate that there are specific stages of induced chondrocyte differentiation in this experimental system. Third, this report includes a new study, in which DBP-regulated genes were used as a data base to derive new information on the cell biology of chondrocytes. The objective was to determine whether a set of genes expressed during induction of chondrocyte differentiation is also expressed by mature articular chondrocytes. Our search of the literature for 59 of the DBP-regulated genes disclosed that expression of 20 of them (33%) had been documented in mature cartilage or chondrocytes. Of the 39 genes not previously documented in cartilage, 11 were tested by RT-PCR and all were found to be expressed in freshly isolated adult human chondrocytes. This review and these new data show how the strategy of high-throughput methods and functional data reduction can expand our knowledge of chondrocyte cell biology. PMID- 14745235 TI - Regulation of cranial suture morphogenesis. AB - The cranial sutures are the primary sites of bone formation during skull growth. Morphogenesis and phenotypic maintenance of the cranial sutures are regulated by tissue interactions, especially those with the underlying dura mater. Removal of the dura mater in fetuses causes abnormal suture development and premature suture obliteration. The dura mater interacts with overlying tissues of the cranial vault by providing: (1) intercellular signals, (2) mechanical signals and (3) cells, which undergo transformation and migrate to the suture. The intercellular signaling governing suture development employs the fibroblast growth factors (FGFs). In rats during formation of the sutures in the fetus, FGF-1 is localized mainly in the dura mater, while other FGFs are expressed in the overlying tissues. By birth, FGF-2 largely replaces FGF-1 in the dura mater. FGFs present in the calvaria bind either the IIIb or IIIc mRNA splice variants of the FGF receptors (FGFRs) 1, 2, or 3. Monoclonal antibodies to the b variant of FGFR2 were used to determine the distribution of FGFR2IIIb during suture development and its extracellular localization. FGFR2IIIb is present in association with mature osteoblasts and osteogenic precursor cells of the suture in the fetus. Ectodomains of FGFR2IIIb, the products of proteolytic cleavage of the receptors, were present throughout the extracellular matrix of sutures resisting obliteration (coronal and sagittal), but absent from the core of sutures undergoing normal fusion (posterior intrafrontal). This observation is consistent with a possible mechanism, in which truncated receptors bind FGFs, thus regulating free FGF available to nearby cells. Mechanical signaling in the calvaria results from tensional forces in the dura mater generated during rapid expansion of the neurocranium. Posterior intrafrontal sutures of rats, which fuse between days 16 and 24, were subjected to cyclical tensional forces in vitro. Significant delay in the timing of suture fusion and increases in the expression domains of FGFR1 and 2 were observed, demonstrating the sensitivity of suture patency to mechanical signals and a possible role of the FGF system in mediating such stimuli. Finally, cells of the dura mater beneath the intrafrontal and sagittal sutures were observed to undergo a morphological transformation to a dendritic morphology and migrate into the suture mesenchyme between days 10 and 16 of development. This process may participate in suture and bone morphogenesis and influence the patency of the sutures along the anterior-posterior axis. PMID- 14745236 TI - Forming a multinucleated cell: molecules that regulate myoblast fusion. AB - In mammals, cell fusion occurs among a limited number of cell types: sperm and oocytes during fertilization, trophoblasts during placenta formation, macrophages during giant cell and osteoclast formation and myoblasts in the formation of myofibers and myotubes. The molecular mechanisms involved in these membrane fusion events largely are unknown. This review will focus on the known molecules that regulate myoblast fusion with an emphasis on a novel signaling pathway involving the calcium-regulated transcription factor NFATC2 in the regulation of myoblast fusion. PMID- 14745237 TI - Perlecan functions in chondrogenesis: insights from in vitro and in vivo models. AB - Perlecan is a large heparan sulfate proteoglycan that is typically found in basal lamina of adult and embryonic tissues. Recent studies have demonstrated that perlecan accumulates impressively during cartilage development and is maintained as the major heparan sulfate proteoglycan of adult cartilage. In vertebrates, perlecan mutations result in skeletal defects. Moreover, in vitro studies indicate that perlecan can stimulate early stages of cartilage differentiation and cooperate with chondrogenic growth factors to promote this process. This short article will summarize these results and propose a model for perlecan function that incorporates these genetic and cell biological findings. PMID- 14745238 TI - Elevated vascular endothelial growth factor in keloids: relevance to tissue fibrosis. AB - Excessive scar or keloid shares common features of a benign dermal growth. Yet, in contrast to malignant tumor, a keloid does not expand beyond the dermis. What triggers the continuing growth of a benign lesion? Deficient or overabundant levels of vascular endothelial growth factor have been reported to contribute to impaired or excessive wound healing. Although numerous studies have examined the pathophysiology of impaired wounds, little information has been provided on mechanisms of exuberant healing. The molecular basis of keloid formation is governed by the interplay of cellular signaling pathways, specific target gene activation, and the nature of the microenvironment. Recent works have demonstrated an accumulation of hypoxia-inducible factor-1alpha protein in freshly biopsied keloid tissues, thus providing first evidence that a local state of hypoxia exists in keloids. Our findings and the findings of others support at least two plausible mechanisms implicated in the development of fibrotic wounds, a state of ongoing fibroplasia or inflammation and an excessive accumulation of extracellular matrix. This article will review recent works examining the potential role of vascular endothelial growth factor in keloid pathogenesis with particular focus on its involvement in the two proposed pathological processes, a prolonged inflammation and an altered balance in extracellular matrix metabolism. PMID- 14745239 TI - Gene therapy approaches for bone regeneration. AB - Gene therapy represents a promising approach for delivering regenerative molecules to specific tissues including bone. Several laboratories have shown that virus-based BMP expression vectors can stimulate osteoblast differentiation and bone formation in vivo. Both in vivo and ex vivo transduction of cells can induce bone formation at ectopic and orthotopic sites. Adenovirus and direct DNA delivery of genes encoding regenerative molecules can heal critical-sized defects of cranial and long bones. Although osteogenic activity can be demonstrated for individual BMP vectors, substantial synergies may be achieved using combinatorial gene therapy to express complimentary osteogenic signals including specific combinations of BMPs or BMPs and transcription factors. Further control of the bone regeneration process may also be achieved through the use of inducible promoters that can be used to control the timing and magnitude of expression for a particular gene. Using these types of approaches, it should be possible to mimic natural processes of bone development and fracture repair and, in so doing, be able to precisely control both the amount and type of bone regenerated. PMID- 14745242 TI - Development of a tissue-engineered human oral mucosa: from the bench to the bed side. AB - The main objective of this publication is to make the reader aware of the complexity and steps that are necessary to make a Food and Drug Administration (FDA)-approved laboratory produced cell-based device, for use in clinical trials for reconstructive surgery. Most tissue-engineered cell-based devices are considered as 'human somatic cell therapy' and fall under the auspices of the Center of Biologic Evaluation and Research (CBER) and are considered a combination product by the FDA. We have illustrated the algorithm that is necessary to follow an Independent New Drug (IND) application by using our ex vivo produced oral mucosa equivalents (EVPOME), a tissue-engineered oral mucosa, as an example of a cell-based device that needs FDA approval prior to clinical application. By illustrating the experimental approach and presenting resulting data we attempt to explain each step that we address along the way. PMID- 14745241 TI - New insight into progenitor/stem cells in dental pulp using Col1a1-GFP transgenes. AB - In recent years there has been increasing progress in identifying stem cells from adult tissues and their potential application in tissue engineering. These advances provide a promising future for tooth replacement/regeneration. Essential for this approach is the identification of donor stem cells for various components of the teeth. Our studies show that pOBCol3.6GFPtpz and pOBCol2.3GFPemd transgenic animals provide a unique model to gain insight into stem cells in the dental pulp. Our in vivo studies of the developing teeth of these transgenic lines show both Col1a1-GFP transgenes are expressed in functional and fully differentiated odontoblasts. The patterns of expression of Col1a1-GFP transgenes during odontoblast differentiation correlates with the expression of DSPP. In the developing craniofacial bones both Col1a1-GFP transgenes are also expressed in osteoblasts and osteocytes of alveolar and calvarial bones. In the alveolar bones, the expression of Col1a1-GFP in osteocytes correlates with the expression of DMP1. Col1a1-3.6-GFP is expressed in the entire layer of the periosteum and in suture mesenchyme containing osteoprogenitor cells. On the other hand, Col1a1-2.3- GFP expression was limited to the osteoblastic layer of the periosteum and was not detected in the fibroblastic layer of the periosteum or in the suture mesenchyme. These observations indicate that Col1a1-3.6-GFP and Col1a1-2.3-GFP transgenes identify different subpopulations of cells during intramembranous ossification. By using the coronal portion of dental pulps isolated from postnatal transgenic mice our observations also provide direct evidence that the dental pulp contains progenitor/stem cells capable of giving rise to a new generation of odontoblast like cells, as well as osteoblast-like cells. PMID- 14745243 TI - Optimal degradation rate for collagen chambers used for regeneration of peripheral nerves over long gaps. AB - The experimental study of peripheral nerve regeneration has depended heavily on the use of a nerve chamber in which the stumps of the transected nerve are inserted. A large variety of chamber fillings and chamber types have been used in an effort to induce a higher quality of regeneration across the gap initially separating the two stumps. In this study we studied the morphology of nerves regenerated across a 15 mm gap following implantation of a series of five chambers. The chambers were fabricated from type I collagen and possessed identical pore volume fractions as well as average pore diameters, but differed in cross-link density continuously along the series. The residual mass of the implanted chambers at 9 weeks was observed to increase continuously with increasing cross-link density along the series, indicating a continuous decrease in degradation rate. The quality of regenerated nerves, determined by the number of large diameter fibers (A-fibers) per nerve, the average diameter of all axons and the ratio of area occupied by axons (N-Ratio), was superior at an intermediate level of chamber degradation rate. The maximal quality of peripheral nerve regeneration corresponded to an optimal degradation rate with an estimated chamber half-life of approximately 2-3 weeks following implantation. A speculative mechanistic explanation of the observed optimum focuses on the hypothetical role of cell and cytokine traffic that may take place through holes in the chamber generated by the degradation process. The data show the presence of a hitherto unreported optimal chamber degradation rate that leads to regenerated nerves of maximum quality. PMID- 14745240 TI - Different effects of BMP-2 on marrow stromal cells from human and rat bone. AB - Bone morphogenetic proteins (BMPs) promote the differentiation of osteoprogenitor cells, and also induce osteogenesis in bone marrow stromal cells (MSC) from rats and mice. However, compared to results with animal models, BMPs are relatively inefficient in inducing human MSC to undergo osteogenesis, and are much less effective in promoting bone formation in human clinical trials. Previous studies indicated that, while human MSC respond to dexamethasone with elevated levels of the osteoblast marker alkaline phosphatase, most isolates of human MSC fail to show alkaline phosphatase induction in response to BMP-2, BMP-4, or BMP-7. Several other genes known to be induced by BMPs are appropriately regulated; thus, human MSC are capable of some BMP-activated signaling. Analysis of the BMP receptors ALK-3 and ALK-6 indicated that, although ALK-6 mRNA was not expressed in human MSC, overexpressing a constitutively active ALK-6 receptor did not induce elevated alkaline phosphatase. Real-time RT-PCR was used to investigate expression of several osteoblast-related transcription factors in MSC after 6 days' exposure to BMP2 or dexamethasone. Msx-2, a transcription factor that has been reported to inhibit differentiation of osteoprogenitor cells, showed 10-fold elevation in BMP-2-treated human MSC, but not in BMP-2-treated rat MSC. Overexpression of Msx-2 in human and rat MSC, however, did not alter alkaline phosphatase levels, which suggests that absence of BMP-stimulated alkaline phosphatase was not caused by the BMP-2-induced increase in Msx-2. Although Runx2 isoforms have been implicated in control of osteoblast differentiation, levels of this transcription factor were unaffected by BMP treatment. Expression of the FKHR transcription factor, which has been reported to regulate alkaline phosphatase transcription in mouse cells, showed a modest increase in response to BMP-2, but a much greater increase in dexamethasone-treated cells. We propose that BMP regulation of the bone/liver/kidney alkaline phosphatase gene is indirect, requiring expression of new transcription factor(s) that behave differently in rodent and human MSC. PMID- 14745245 TI - [Cancer metastasis and metastasis suppressors]. AB - Cancer metastasis, a complex and sequential network of cellular events involved in the migration and establishment of malignant cells from original site to distant foci, is an important and significant contributor to morbidity and mortality of cancer patients. Despite the clinical importance of cancer metastasis, its molecular and biochemical mechanism remains unclear. The identification of tumor suppressor gene confirmed that metastasis might involve the functional loss of genes that maintain the cellular differentiation optimally. Metastasis suppressor is defined by the ability to reduce the metastatic property of cancer cells without affecting its tumorigenesis. Since NM23 was first identified in 1988 as a metastasis suppressor, several metastasis suppressor genes have been identified and characterized. In this article, we review the complex and multi-step process of cancer metastasis and describe the recent progress of metastasis suppressors in the studies of identified. Consequently, we hope to introduce the new therapeutic target for the metastasis suppressors in cancer patients. PMID- 14745244 TI - Transitive behavior in hippocampal-lesioned pigeons. AB - The hippocampus of birds and mammals is critical for the learning of map-like memory representations of environmental space. It has been suggested that the hippocampus of rats also participates in non-spatial relational learning, including the learning of non-spatial transitive relationships among odor stimuli [Bunsey and Eichenbaum, Nature 1996]. Although transitive-like learning has been demonstrated in a variety of vertebrate species, from a comparative perspective the role of the hippocampus in this form of learning has not been tested in other amniote groups. We trained control and hippocampal-lesioned homing pigeons on a series of visual, non-spatial, go/no-go conditional discriminations and then tested them on novel transitivity probe trials. The hippocampal-lesioned pigeons were as successful as control pigeons in responding appropriately to correct and incorrect transitivity pairs. The finding that the homing pigeon hippocampal formation is not necessary for solving this serial, conditional discrimination task is important for further understanding hippocampal function across species, and represents one of the few studies that have attempted to localize a brain region responsible for the phenomenon of transitive behavior learning. PMID- 14745246 TI - [Clinical characteristics at diagnosis and course of Korean patients with Crohn's disease]. AB - BACKGROUND/AIMS: The aim of this study was to evaluate the clinical characteristics and course of Korean patients with Crohn's disease (CD). METHODS: A total of 113 patients who were newly diagnosed as having CD at the Asan Medical Center between October 1989 and December 2001 were included. RESULTS: The male to female ratio was 1.9:1 and the mean age at diagnosis was 24.2 +/- 7.8 years. The disease location was the small bowel in 19.5%, the large bowel in 11.5%, and the both small and large bowels in 69.0% of the patients. Empirical anti-tuberculous medications were administered to 53 cases (46.9%). Major complications of CD were intestinal obstruction (25.7%) and perianal fistula (42.5%). The cumulative remission rate was 75.7% at 1 year and 80.7% at 3 years from diagnosis. The cumulative relapse rate was 30.7% at 1 year and 53.0% at 3 years from remission. The cumulative operation rate was 11.5% at 1 year and 13.1% at 3 years from diagnosis. CONCLUSIONS: There are no differences in the remission and recurrence rates between Korean and Western patients with CD. Although the cumulative operation rate seems to be lower in Korea than in Western countries, the cause of the difference is uncertain. Further studies are needed to determine the characteristics of CD in Korea. PMID- 14745247 TI - [Serotonin transporter gene polymorphism in healthy adults and patients with irritable bowel syndrome]. AB - BACKGROUND/AIMS: Serotonin is thought to be an important neurotransmitter in the pathogenesis of irritable bowel syndrome (IBS). It is reported that functional polymorphism in the promotor region of the serotonin transporter gene is related with the subtypes of IBS and shows racial difference. However, a functional relation between polymorphism and IBS is not clear. The aim of this study was to investigate 5-hydroxytryptamine transporter (5-HTT) gene polymorphism in patients with IBS. METHODS: For fifty-six healthy controls and 33 patients with IBS fulfilling Rome II criteria, 5'-flank promotor region of 5-HTT gene was analyzed by polymerase chain reaction. RESULTS: The genotypes of healthy controls were S/S (57.1%), S/L (37.5%), and L/L (5.4%). Those of IBS patients were S/S (54.5%), S/L (36.4%), and L/L (9.1%). IBS patients were divided into three groups: diarrhea predominant (n=15; S/S, 40%; S/L, 53.3%; L/L, 6.7%), constipation predominant (n=12; S/S, 75.0%; S/L, 8.3%; L/L, 16.7%), diarrhea-constipation alternating type (n=6; S/S, 50%; S/L, 50%). There was no statistical difference in the 5-HTT gene polymorphism between patients and controls, and according to the subtypes of IBS patients (p=0.135). CONCLUSIONS: There was no relationship between serotonin transporter gene polymorphism and IBS. However, allele S/S genotype was most prominent genotype in both controls and patients. PMID- 14745248 TI - [Prognostic factors of stage II rectal cancer]. AB - BACKGROUND/AIMS: We aimed to verify the prognostic factors of stage II rectal cancer and the effect of radiation therapy on the survival and local recurrence rate. METHODS: This study was undertaken in 202 patients who underwent curative resection of rectal cancer and confirmed to be stage II between July 1989 and December 1996. Univariate and multivariate (Cox's model) analyses of survival were employed to identify prognostic factors. Statistical significance was assigned by p value of <0.05. RESULTS: Overall recurrence occurred in 32 patients. Four patterns of recurrence were observed: hematogenous recurrence in 17 patients, local recurrence in 11, peritoneal seeding in two and simultaneous hematogenous and local recurrence in two cases. Overall 5-year survival rate was 85.6% and 5 year disease free survival rate was 82.8%. There was no significant difference in local recurrence rate and survival according to radiation therapy or location of cancer. In multivariate analysis, the number of harvested lymph node was only a prognostic factor. CONCLUSIONS: The number of harvested lymph nodes has prognostic value in stage II rectal cancer. Postoperative radiation therapy should be considered for stage II rectal cancer with poor prognostic factors although radiation did not decrease local recurrence rate in present study. PMID- 14745250 TI - [Clinical characteristics of pancreatic cancer according to the presence of diabetes mellitus]. AB - BACKGROUND/AIMS: Pancreatic cancer is fatal with a dismal 6-month median survival from diagnosis. Diabetes mellitus is reported to be present up to 33.3 percent of patients with pancreatic cancer. The reason for the high frequency of diabetes is unknown. We studied the prevalence and duration of diabetes in patients with pancreatic cancer and the relationship between the two diseases. METHODS: A total of 152 patients with pancreatic cancer diagnosed at Yeungnam University Hospital from January 1999 to December 2001 were enrolled in this study. Clinical features, family history, smoking history, and characteristics of the tumor were compared between diabetic and non-diabetic groups. RESULTS: Among 152 patients with pancreatic cancer, 43 patients (28.3%) had diabetes. In diabetic group, mean age of diagnosis was significantly younger than non- diabetic group (62.0 +/- 7.2 vs. 65.0 +/- 8.8, p<0.05). Most of the patients with diabetes had non-insulin dependent diabetes mellitus (NIDDM) and did not have family history. Diabetes was diagnosed within 2 years after the diagnosis of pancreatic cancer in 35 patients (74.3%) of the diabetic group. There were no differences in the location and stage of tumor, chief complaints, presence of weight loss, and body mass index between the two groups. CONCLUSIONS: Diabetes mellitus occurs frequently in patients with pancreatic cancer and does not influence clinical features of pancreatic cancer. Pancreatic cancer should be suspected in patients with recent onset diabetes, especially in patients without family history of diabetes and with type of NIDDM. PMID- 14745249 TI - [Clinical features and prognostic factors in primary adenocarcinoma of the appendix]. AB - BACKGROUND/AIMS: Primary appendiceal adenocarcinoma is a rare neoplasm that constitutes less than 0.5% of all gastrointestinal neoplasm. The aim of this study was to figure out its clinicopathologic characteristics that are not well understood. METHODS: We reviewed the medical records of nineteen patients (9 males and 10 females) with histologically proven appendiceal adenocarcinoma. They had been treated at Asan Medical Center between June 1989 and December 2002. Their median follow-up duration was 72.5 months. RESULTS: Their median age was 56.5 (range, 33-80) years. Thirteen patients had mucinous variants and the other five had adenocarcinoma. Seven patients (36.8%) were diagnosed as acute appendicitis. In fact, none of the patients was diagnosed correctly before surgery. The operative procedure, included right hemicolectomy in 9 patients, appendectomy alone in 2 patients, and debulking of their tumors or a biopsy in 8 patients. The 5-year survival rate was 20.5%. The patients with mucinous type had better prognosis than those with the non-mucinous type (p<0.01). In the patients with mucinous type, the survival rate after debulking operation was similar to that after right hemicolectomy. CONCLUSIONS: The most important prognostic factor of primary appendiceal adenocarcinoma was histology. The outcome of debulking operation is being watched compared with that of right hemicolectomy in mucinous variant. PMID- 14745251 TI - [Cationic trypsinogen gene mutation in patients with chronic idiopathic pancreatitis]. AB - BACKGROUND/AIMS: Mutation of Cationic trypsinogen gene is clearly associated with hereditary pancreatitis and plays an important role in the pathogenesis of pancreatitis. According to literature, this mutation is occasionally occurred in patients with pancreatitis in Western countries and Japan. The aim of this study was to find out whether the mutation was observed in Korean patients with chronic idiopathic pancreatitis. METHODS: Peripheral blood samples of 11 patients with chronic idiopathic pancreatitis were collected consecutively, and DNA was extracted from the samples. Polymerase chain reaction was performed in exon 2 and 3 of cationic trypsinogen gene. Then, DNA products were purified and sequenced. RESULTS: The mutation was not found in exon 2 and 3 of cationic trypsinogen gene in these patients. CONCLUSIONS: There was no cationic trypsinogen mutation in Korean patients with chronic idiopathic pancreatitis. Further large sampled cohort study is needed. PMID- 14745252 TI - [A case of gangliocytic paraganglioma in duodenum]. AB - Duodenal gangliocytic paraganglioma derived from neural crest is a peculiar neuroendocrine tumor. It is incidentally found during radiographic studies or due to gastrointestinal hemorrhage caused by frequent ulceration of the overlying mucosa. Most lesions are pedunculated and submucosal with distinctive histology consisting of endocrine cells, ganglion cells and spindle-shaped Schwann cells. We experienced a duodenal gangliocytic paraganglioma in a 40-year-old woman presenting with episodes of melena. Esophagogastroduodenoscopy revealed a submucosal ulcerated tumor in the second portion of duodenum and a biopsy confirmed gangliocytic paraganglioma. The tumor was enucleated through a duodenotomy. It's size was 5.5 X 3 X 1.5 cm sized and revealed positive cellular reaction for chromogranin, synaptophysin, neuron-specific enolase, and neurofilament by immunohistochemistry. The majority of the reported duodenal gangliocytic paraganglioma were of benign nature. Therefore, radical surgery or a lymph node dissection could be avoided if that disease was confirmed. However, thorough investigation for lymph node metastasis and postoperative follow-up are needed. We report the first case of a duodenal gangliocytic paraganglioma in Korea with a review of literature. PMID- 14745253 TI - [A case of gastritis cystica profunda associated with high grade tubular adenoma]. AB - Gastritis cystica profunda is usually characterized by hyperplastic and cystic dilatation of pseudopyloric gland with submucosal invasion. These lesions possess malignant histologic features as metaplastic and even dysplastic alteration can be observed, however they usually exhibit benign behavior. Macroscopically, gastritis cystica profunda may present as a submucosal tumor or as a polyp. In rare case, however, it can be found as a giant mucosal fold. The proposed pathogenesis of lesion relates to ischemia, chronic inflammation, and the presence of foreign body. Herein, we report a case of gastritis cystica profunda associated with high grade tubular adenoma in a patient without antecedent surgery. PMID- 14745254 TI - [A case of sarcomatoid combined hepatocellular-cholangiocarcinoma]. AB - Combined hepatocellular-cholangiocarcinoma (HCC-CC) with sarcomatoid features is an extremely rare primary liver cancer, of which only four cases have been reported. We report a case of sarcomatoid combined HCC-CC in a 60-year-old woman who complained of right upper quadrant pain and presented with a 7 cm mass in the S4 region of the liver in abdominal CT. Ultrasonography-guided needle biopsy diagnosed it as HCC, and left lobectomy of the liver followed. Microscopically, the tumor consisted of two portions: HCC portion showing trabecular pattern, which had partially sarcomatous area with spindle-shaped tumor cells, and CC portion with glandular pattern. Immunohistochemically, HCC portion reacted positively with alpha-fetoprotein while CC portion demonstrated positive reactivity with carcinoembryonic antigen and mucicarmine. Sarcomatoid cells reacted positively for cytokeratin. She died of tumor recurrence and hepatic failure 12 months after the operation. Combined HCC-CC has poor prognosis, and sarcomatoid HCC has high metastatic potential and poor prognosis compared with ordinary HCC. PMID- 14745255 TI - [Idiopathic hypereosinophilic syndrome presenting as a single mass in the liver]. AB - Hypereosinophilic syndrome is characterized by persistent blood eosinophilia of 1,500/mm(3) or more in the absence of known causes and multiorgan dysfunction by eosinophil-related tissue damage. In Korea, some cases of hypereosinophilic syndrome with hepatic involvement have been described with prolonged benign clinical courses. Most of them were diffuse or multifocal lesions in imaging studies, and any case presenting as a large single mass lesion has not been described. Herein we report a case of hypereosinophilic syndrome with hepatic involvement in a 48-year-old woman who presented with a giant single mass. By abdominal CT scan, a lobulated well-margined heterogenous mass lesion was detected in the left lateral segment of the liver. Liver biopsy revealed severe eosinophilic infiltration and centrilobular necrosis of hepatocytes. The lesion completely disappeared after steroid administration for eleven months. PMID- 14745256 TI - Malignant gastrointestinal stromal tumor of the ampulla of Vater: a case report. AB - GIST is a rare neoplasm, the majority of GISTs are located in the stomach and small intestine. Most GISTs are diagnosed histopathologically after resection because of submucosal location. A 37-year-old female patient presented with a 2 weeks history of generalized weakness, nausea accompanied by intermittent passage of black, tarry stools. Esophagogastroduodenoscopy and ERCP showed a large round mass measuring 5 cm in diameter in the ampulla of Vater with ulcer crack. Endoscopic multiple biopsies from the mass including ulcer base were taken. Light microscopic findings showed spindle-shaped and epitheloid tumor cells having high cellularity and frequent mitotic figures. On immunohistochemical stainings, the tumor cells were positive for CD34 and smooth muscle actin. Based on these preoperative findings, a diagnosis of malignant GIST of the ampulla of Vater was made probably. After operation, immunohistochemical studies revealed positive reaction for c-kit and vimentin, as well as focally reactive for CD34 and smooth muscle actin. We report a case of GIST in the ampulla of Vater presenting with melena that was diagnosed preoperatively and postoperatively. PMID- 14745258 TI - Breast reconstruction with gluteal artery perforator (GAP) flaps: a critical analysis of 142 cases. AB - Fujino was the first to introduce gluteal tissue as a free flap for breast reconstruction. The use of the musculocutaneous flap from the buttock in breast reconstruction has been championed by Shaw. Despite the initial enthusiasm for this area as a donor site, few other large series exist on the subject. Two decades of experience with this region as a donor site led to recognition of advantages and drawbacks. Furthermore, use of both the superior and inferior gluteal musculocutaneous flap was associated with certain important donor site complications and the use of vein grafts to allow for microvascular anastomosis. The evolution of free tissue transfer has progressed to the level of the perforator flap. This reconstructive technique allows elevation of tissue from any region consisting only of fat and skin. This minimizes donor site morbidity by allowing preservation of the underlying muscle and coverage of important structures in the region such as nerves. The superior and inferior gluteal perforator flaps have been used at our institution for breast reconstruction since 1993. The superior gluteal artery perforator (S-GAP) flap is our preferred method of breast reconstruction when the abdomen is not available or preferable. We report the result of this flap over the past 9 years and point out important surgical refinements, advantages, disadvantages, and lessons learned during this time. PMID- 14745257 TI - Brain cancer and cosmetic breast implants: a review of the epidemiologic evidence. AB - Recently, the findings of 1 mortality follow-up study have raised concern regarding the risk of brain cancer among women with cosmetic breast implants. In this review, the authors examine the results of 4 existing large-scale incidence follow-up studies comprising more than 10,000 women with cosmetic implants followed for as long as 29 years. Overall, there were 12 observed incident cases of brain cancer compared with 9.6 cases expected, yielding a nonstatistically significant summary standardized incidence ratio of 1.25 (95% confidence interval, 0.7-2.2). Based on the confidence interval alone, this result rules out the standardized mortality ratio of 2.45 reported in the 1 positive mortality study. Overall, the epidemiologic evidence does not support an association between breast implants and brain cancer incidence. PMID- 14745259 TI - Suspicious findings in reduction mammaplasty specimens: review of 182 consecutive patients. AB - Breast reduction mammaplasty allows examination of specimens from a seemingly healthy population for the presence of proliferative breast disease. The authors reviewed the charts of all reduction mammaplasty patients of a single surgeon over 7.5 years for age, family history, mammographic results, unilateral or bilateral nature of the procedure, and final pathologic diagnosis. Of 182 patients, 168 had bilateral and 14 had unilateral breast reductions. Ages ranged from 16 to 79 years (average and median: 37 years and 35 years respectively). Fifty-seven patients (31%) were younger than 30 years, 53 patients (29%) were between the ages 30 years and 39 years, for a total of 110 patients (60%) younger than 40 years in this study. A total of 163 patients (89%) had a diagnosis of normal breast tissue. Nineteen patients (10%) had proliferative changes: 9 patients (5%) without atypia, 5 patients (3%) with atypia, 3 patients (2%) with sclerosing adenosis, and 1 patient each (0.5%) with papillomatosis and lobular carcinoma in situ. A total of 95% of patients with proliferative changes were older than 30 years. Women ages 30 to 39 years may be at higher risk (15%) of having proliferative changes than previously reported, and histologic examination of all reduction mammaplasties is recommended. PMID- 14745260 TI - Ultrasound guided minimally invasive breast surgery (UMIBS): a superior technique for gynecomastia. AB - The last 30 years has seen a shift in surgical treatment of breast diseases to less invasive, more conservative treatment options. The mammotome equipment was originally introduced as a diagnostic tool, but advances in technology have extended its role to therapeutic procedures. The mammotome device (8-gauge) is inserted through a cosmetically placed 4-mm scar and breast tissue is resected sequentially using a suction facility without the need to remove the biopsy device. This is done under ultrasound guidance. Operative morbidity and postoperative score for cosmesis and patient satisfaction were recorded prospectively. Patients were reviewed in the clinic after 6 to 8 weeks. Five patients (4 unilateral, 1 bilateral) with idiopathic gynecomastia were treated. Mean age was 41.8 years (range, 16-88 years) with a median procedural time of 32 minutes. No postoperative morbidity was noted and mean cosmetic score was 9/10. The mammotome is an emerging minimal invasive tool that is safe and ensures excellent cosmesis and very high patient satisfaction rates. PMID- 14745261 TI - An anatomic study of nasal tip supporting structures. AB - Successful rhinoplasty depends on nasal tip support and its influence on nasal tip projection. It is generally agreed that the components of nasal tip support include the attachment between the upper and lower lateral cartilages, the attachment between the lateral crus of the lower lateral cartilage and the pyriform aperture, the attachment between the paired domes of the lower lateral cartilages, and the medial crural attachment to the caudal septum. However, these structures are still not clearly determined, and there was no anatomic study of nasal tip supporting structures in Asia. The purpose of this study was to determine the nasal tip supporting structures and find out the differences in these structures between white and Asian people. Ten noses of fresh cadavers were investigated. Dissection was performed and the previously mentioned nasal tip supporting structures were observed and excised. Histologic examination was done with hematoxylin and eosin stain and Van Gieson elastin stain. Macroscopic study showed that there were dense fibrous tissue between the upper and lower lateral cartilages, dense fibrous tissue and sesamoid cartilages between the lateral crus and the pyriform aperture, loose connective tissue between the paired domes of lower lateral cartilages, and no identified specific tissue between the medial crus and the caudal septum. Microscopic investigation allowed a more detailed analysis of these structures. Between the upper and lower lateral cartilages, dense collagen fibers were running in one direction and anchoring firmly to each cartilage, which meets the histologic criteria of a ligament. Between the lateral crus and the pyriform aperture, there were intermingled collagen fibers and muscular fibers, which meets the histologic criteria of fibromuscular tissue. Between the paired domes of lower lateral cartilages, there were few fibers with abundant amorphous ground substances, which meets the histologic criteria of loose connective tissue. Based on our results, we recommend that the previously mentioned nasal tip supporting structures should be named intercartilaginous ligament, sesamoid fibromuscular tissue, and interdomal loose connective tissue, respectively. In addition, we consider that the loose connection between the domes of middle crura and the absence of an attachment of the medial crura to the caudal septum can be one of the reasons why the nasal tip of Asian people is broad and unprojected and the base is wide. PMID- 14745262 TI - Innervation of the corrugator supercilii muscle. AB - The frowning forehead is unwanted, generally because it gives an impression of anger and displeasure. The frown is formed mainly by the repeated contraction of the corrugator supercilii muscle. We studied the relevant nerve supply to the muscle in detail to enhance selective neurotomy. A bicoronal incision extending downward to the preauricle was made and the subgaleal dissection proceeded in 19 hemifaces of cadavers. The temporal branch of the facial nerve was divided into 2 to 4 smaller branches at the zygomatic arch. There were commonly 3 branches. The temporal branch is located 10 mm lateral to the supraorbital notch or foramen 2.8 to 25 mm above the supraorbital rim. It contains 4 to 7 thin rami (5 on average). A plexus mainly from the inferior ramus partly from the middle ramus of the temporal branch of the facial nerve enters the corrugator supercilii muscle in the supraorbital area. Because the temporal branch had multiple interconnection among its own rami, selective blocking of the middle ramus cannot promise the elimination of the frown line. PMID- 14745263 TI - One-stage reconstruction of infected cranial defects using a titanium mesh plate enclosed in an omental flap. AB - Cranial osteomyelitis is a common complication of brain surgery. A one-stage reconstructive repair of the cranial bone is desirable, but risks active infection. The authors have developed a reconstructive technique for osteomyelitis-related cranial defects that fills the dead space with an omental flap and uses a titanium mesh plate for the structural element. This report describes the successful treatment of 8 patients using this approach. PMID- 14745264 TI - Recipient vessel analysis for microvascular reconstruction of the head and neck. AB - The selection of recipient vessels that are suitable for microvascular anastomosis in the head and neck region is one of many components that is essential for successful free tissue transfer. The purpose of this study was to evaluate a set of factors that are related to the recipient artery and vein and to determine how these factors influence flap survival. A retrospective review of 102 patients over a 5-year consecutive period was completed. Indications for microvascular reconstruction included tumor ablation (n = 76), trauma (n = 13), and chronic wounds or facial paralysis (n = 13). The most frequently used recipient artery and vein included the facial, superficial temporal, superior thyroid, carotid, and jugular. Various factors that were related to the recipient vessels were analyzed and included patient age, recipient artery and vein, diabetes mellitus, tobacco use, the timing of reconstruction, the method of anastomosis, previous radiation therapy, creation of an arteriovenous loop, and use of an interposition vein graft. Successful free tissue transfer was obtained in 97 of 102 flaps (95%). Flap failure was the result of venous thrombosis in 4 and arterial thrombosis in 1. Statistical analysis demonstrated that anastomotic failure was associated with an arteriovenous loop (2 of 5, P = 0.03) and tobacco use (3 of 5, P = 0.03). Flap failure was not related to patient age, choice of recipient vessel, diabetes mellitus, previous irradiation, the method of arterial or venous anastomosis, use of an interposition vein graft, or the timing of reconstruction. PMID- 14745266 TI - Large and giant congenital pigmented nevi of the upper extremity: an algorithm to surgical management. AB - The timing and choice of treatment of congenital giant pigmented nevi continues to evolve under the influence of changing opinions regarding the risk of malignant degeneration and the impact of excision and reconstruction on the affected child. Many studies exist to support a notable enough risk of malignancy to warrant excision, yet other series and pigmented lesion clinics suggest that the risk of malignancy does not warrant the potential scarring and deformity that has followed the surgery necessary to remove these giant lesions. To satisfy both sides in this controversy, we have been challenged to modify our surgical techniques in a manner that minimizes the risk of malignant degeneration and at the same time provides optimal functional and aesthetic outcomes for these complex reconstructions. Thirty consecutive patients with large and giant nevi of the upper extremity were treated over a 23-year period (1979-2002) by the senior author. These patients represent a subset of 259 children (12%) with large or giant congenital pigmented nevi treated and followed during this period of time. In proximal upper extremity lesions, expanded transposition flaps from the upper back and shoulder have effectively eliminated contour defects or circumferential constriction in the upper arm and axilla. An expanded free transverse rectus abdominis musculocutaneous flap has offered a possible avenue for larger lesions (shoulder and upper extremity to below the elbow), and pedicle flaps from the flank (both expanded and nonexpanded) have offered ways of improving the long term contour in the forearm. Expanded and nonexpanded full-thickness skin grafts were chosen for reconstruction of the hand and the fingers. The authors describe in detail the surgical strategies and the techniques for reconstruction of each region of the upper extremity and then bring these ideas together in an algorithm for assessment and treatment of these challenging lesions. PMID- 14745267 TI - Technical modifications and extended applications of the distally based adipofascial flap for dorsal digital defects. AB - A series of 15 cases of distally based adipofascial flaps to cover dorsal digital defects is presented. All flaps were raised just superficial to the dorsal veins (and hence preserving some fat with the reflected flaps) rather than raising them at the subdermal plane. In 3 cases where there was another injury proximal to the defect, which precluded the use of an adipofascial turnover flap from the injured digit, a distally based cross-finger adipofascial flap was used from the adjacent finger. In 4 cases, the flap was used to cover compound fractures. The results showed complete survival of all flaps without loss of the overlying skin graft and without epidermolysis of the donor skin, indicating that raising the flap just superficial to the dorsal veins does not affect the reliability of its blood supply, and yet it enhances the blood supply of the reflected skin flaps. The series also showed uneventful healing of compound fractures covered by the flap, indicating flap reliability to cover exposed fractures. Finally, the study demonstrated the feasibility of using the flap as a cross-finger flap. PMID- 14745268 TI - Proximal radial compression neuropathy. AB - Proximal radial nerve compression occurs infrequently and is diagnosed successfully even less frequently. A large clinical series of patients with proximal radial nerve compression neuropathy was reviewed to determine better the common symptoms, physical findings, and electrodiagnostic findings, and to identify the predictors of better or worse outcome after surgical decompression. Seventy-nine proximal radial compression neuropathies were treated in 71 patients by the same surgeon between 1991 and 2000. The most consistent symptoms were deep aching pain in the forearm, pain radiation to the neck and shoulder, and a "heavy" sensation of the affected arm. The most common physical findings were tenderness over the radial nerve at the supinator muscle level, pain on resisted supination, and the presence of a Tinel sign over the radial forearm. Electrophysiologic studies were of limited value in diagnosis, with 90% of patients having normal findings. On operation, prominent pathology of the posterior interosseous nerve was observed in 36 of 79 limbs (46%). Follow-up ranged from 12 to 86 months (mean, 21 months) with no significant complications or recurrence of symptoms. Of the 79 nerve decompressions, 77% had excellent recovery and 20% were judged to be good. Of 69 patients employed when treated, 60 resumed gainful employment, including 53 who returned to their regular jobs. Proximal radial compression neuropathies are uncommon but present with a basic constellation of symptoms and physical findings, and decompression can provide excellent relief of symptoms. PMID- 14745270 TI - The medial sural MEDIAL GASTROCNEMIUS perforator free flap: an 'ideal' prone position skin flap. AB - The medial sural MEDIAL GASTROCNEMIUS perforator flap is a potentially large, thin cutaneous flap that encompasses the calf skin territory. Its medial sural vascular pedicle has a long leash of large caliber that simplifies microanastomoses when used as a free flap. Because the identification of the requisite perforators and their subsequent intramuscular dissection is facilitated with the patient in a prone position, this can be an "ideal" skin free flap for the posterior aspect of the body. This is especially true for the lower limb where all surgical morbidity could then be restricted to the ipsilateral extremity. This approach has been used in 5 clinical cases, with success except once when the flap was aborted as a result of anatomic anomalies, which is always a concern with muscle perforator flaps. PMID- 14745271 TI - Acellular cadaveric dermis (AlloDerm): a new alternative for abdominal hernia repair. AB - AlloDerm is an intact human tissue matrix with the critical biochemical and structural components of the dermal extracellular scaffold that enable it to recellularize and revascularize. Its biologic nature provides AlloDerm with an improved capacity to reintegrate with surrounding tissues and less inclination toward infection, erosion, extrusion, and rejection compared with synthetic tissue replacement materials. This retrospective review describes the experience with AlloDerm for incisional hernia and transverse rectus abdominis musculocutaneous flap reconstructions at a plastic surgery practice that handles complex, nonprimary repairs. AlloDerm was observed to provide exceptional safety and tolerability, and to become reintegrated and vascularized with surrounding tissue for tolerable and persistent tissue replacement. PMID- 14745272 TI - Donor site morbidity after serratus anterior free muscular flap: a prospective clinical study. AB - This study was designed to assess donor site morbidity after using the serratus anterior muscle as a free vascularized flap. The 3 distal slips of the serratus anterior were harvested from 3 dominant and 4 nondominant shoulders of 7 consecutive patients (age range, 36-61 years) to treat chronic osteitis or infected nonunions of the lower limb. Both donor and recipient sites healed primarily in all patients. Six of the 7 patients were enrolled in a postoperative shoulder-strengthening program. Preoperative and 3-month follow-up Constant scores and peak torque values of the operated shoulders were compared using the Wilcoxon matched pairs signed rank test. The pre- and postoperative Constant score were 95% and 93% respectively. Peak shoulder torque for abduction, adduction, flexion, and extension was assessed at both 60 degrees /second and 120 degrees /second. No statistical differences were found between the pre- and postoperative values. At the final follow-up (mean, 17 months), clinical examination revealed no scapular winging in all patients. Six patients answered a self-administered questionnaire to assess function of the shoulder. The average score in the pain domain was 36.3 points (with 40 points meaning free of pain). The average score in the activities of daily living domain was 18.5 of 20 points. The overall satisfaction rate with the donor site was very good in 2 patients and good in 4 patients. In conclusion, removal of the 3 distal slips of the serratus anterior for use as a free vascularized transfer did not impair shoulder function in this group of patients. PMID- 14745273 TI - The prepuce and circumcision: dual application as a graft. AB - Circumcision is probably one of the first plastic surgery operations that has been used for centuries. The aim of this study was to apply the bilamellar tissue (skin and mucosa) obtained from circumcision to various defects and to evaluate the clinical results. During the last 2 years, 19 patients have been operated, and the skin and mucosal grafts were applied individually or simultaneously. The etiology was trauma for the whole series of patients (12 burns and 7 strap injuries). In 15 patients the defect was localized to the hand whereas in 4 patients it was located on the dorsum of the foot. In 10 patients, mucosa and skin graft were applied to the same defect as a single, compact layer. In 9 patients, skin and mucosa were applied separately to multiple defects. Using these methods, comparative evaluation of the consequences of prepuce mucosal and skin graft applications could be made. Four obvious differences were observed: (1) in mucosal grafts, early graft edema that resolves spontaneously after 48 hours; (2) better adaptation of the mucosal grafts to the recipient bed; (3) hyperpigmentation in both graft types, but the skin part was slightly darker than the mucosa; and (4) less secondary contraction was seen in mucosal grafts. The results were evaluated in light of the authors' knowledge of the prepuce as an alternative full-thickness donor site. The relative differences in the dual anatomic structure of mucosa and skin, and the role of circumcision as a medical, cultural, and religious application in some societies are discussed. PMID- 14745274 TI - Integrin-linked kinase: a possible role in scar contracture. AB - Integrin-linked kinase (ILK) participates with beta1 integrin to mediate extracellular matrix interactions, such as extracellular matrix reorganization. Thus, ILK is hypothesized to influence wound contraction and scar contracture and, as such, would be a target molecule to manipulate pharmacologically in expediting wound contraction or possibly preventing scar contracture. The expression of ILK messenger ribonucleic acid, along with ILK-protein expression, was found in fibroblasts. The localization of ILK in human skin and rat granulation tissue was documented by immunohistology. ILK was present in human dermal fibroblasts, but was not found in human epidermal cells in skin. Cells were transfected with wild-type ILK or kinase-deficient ILK (E359K) and were assayed for collagen lattice contraction, migration, and myosin adenosine triphosphatase (ATPase) activity. Cells overexpressing E359K were poorer at collagen lattice contraction than control cells, whereas cells overexpressing wild-type ILK were equal to control cells at lattice contraction. ILK overexpression enhanced cell migration, but E359K overexpression did not affect cell migration. Neither ILK nor E359K overexpression altered myosin ATPase activity. Hence, ILK action within fibroblasts appears unrelated to myosin ATPase control of microfilament-generated forces. ILK appears to be a target molecule for pharmacologic manipulation to expedite wound contraction or to prevent scar contracture. PMID- 14745275 TI - Forearm musculofasciocutaneous flap to cover glenohumeral arthrodesis hardware during reconstruction of the flail upper extremity. AB - SUMMARY: For selected patients with flail upper extremities after brachial plexus injury, glenohumeral arthrodesis, above-the-elbow amputation, and fitting of an above-the-elbow prosthesis are effective reconstructive options. In such situations, soft-tissue thinning at the shoulder, especially deltoid atrophy, presents the potential problem of shoulder fusion hardware causing pain or even eroding overlying soft tissue. The authors have used a pedicled forearm musculofasciocutaneous flap, elevated just before above-the-elbow amputation and subsequently transposed to the shoulder, to provide high-quality soft-tissue coverage of the arthrodesis hardware. Preliminary results with three patients suggest that this procedure may be useful for preventing tenderness and breakdown of tissue overlying glenohumeral arthrodesis hardware. In all three patients, the forearm flap remained well perfused, and all wounds healed fully with no subsequent problems with skin breakdown. All three patients had long-term benefit from prosthesis fitting. PMID- 14745276 TI - Severe neck scar contracture reconstructed with a ninth dorsal intercostal perforator augmented "Super-Thin Flap". AB - The authors have reconstructed neck scar contractures with "super-thin flaps" (subdermal vascular network [SVN] flaps) since 1994 and have used the circumflex scapular artery and veins (CSAV) and dorsal intercostal perforators (DICPs) to augment the blood flow in the distal portion of the flaps. These free microvascular augmentations enlarge the flap survival area. In this report, the authors describe a severe neck scar contracture reconstructed with a ninth DICP augmented "super-thin flap." The patient was a 51-year-old woman with severe flame burns on 44% of her total body surface area, resulting from a cooking accident. After emergency skin grafting, the patient had a severe scar contracture and intractable ulcer of the anterior neck. CSAV and a ninth DICP augmented occipito-cervico-dorsal (OCD) "super-thin flap" transfer were used to reconstruct the anterior neck. The flap size was 28 x 15 cm, and it survived completely. The cervico-mental angle was clear and esthetically pleasing, and not only the aesthetic results but also the functional results were excellent. PMID- 14745278 TI - And still more bloopers. PMID- 14745277 TI - Surgical management of hidradenitis suppurativa of the nipple-areolar complex. AB - Involvement of the nipple-areolar complex is an infrequent area of distribution of hidradenitis suppurativa. The difficulty of this particular distribution has been encountered in the past and resulted in bilateral mastectomies. The authors describe successful treatment of a patient with hidradenitis suppurativa of the breasts and nipple-areolar complexes using breast-conserving measures. Normal breast contour was preserved and overall cosmetic result was good. PMID- 14745280 TI - Endoscopic therapy with multiple plastic stents for benign biliary strictures due to chronic calcific pancreatitis: the good, the bad, and the ugly. PMID- 14745281 TI - Pitfalls of endoscopic ultrasound staging after application of electrocautery. AB - Patients may be referred for endosonography after endoscopic resection of polyps because of cancer identified in the histologic specimen. To assess the effects of electrocautery-induced tissue changes on tumor staging by endosonography, endosonography findings after endoscopic removal of large polyps were correlated with surgical and endoscopic pathology. Endosonography findings revealed irregular and thickened wall layers, especially in the muscularis propria with pseudopod extensions. Five of 7 patients had evidence of cancer in the endoscopic specimen. However, no residual tumor was found in the surgically resected bowel (2 patients) or in subsequent biopsies of the endoscopic resection site (3 patients). In 2 other patients, no cancer was present in the endoscopic specimen, and follow-up biopsies of the endoscopic resection site were all benign. Electrocautery-induced inflammatory changes create hypoechoic changes within the gut wall that may mimic tumor invasion. Irregularities in the muscularis propria layer cannot be relied upon to diagnose a T2 or T3 lesion by endosonography in this setting. Patients with large polyps greater than 2 cm and other mucosal lesions with malignant potential should undergo endosonography prior to endoscopic resection. PMID- 14745282 TI - Gastrointestinal endoscopic evaluation of premenopausal women with iron deficiency anemia. AB - GOALS: To evaluate whether the gastrointestinal tract could be a source of chronic blood loss in premenopausal women with iron deficiency anemia. BACKGROUND: While premenopausal women with iron deficiency anemia are typically managed with simple iron replacement, the standard of care for postmenopausal women and men is to exclude a gastrointestinal source of bleeding. STUDY: We identified 111 premenopausal women who underwent endoscopy for the sole indication of iron deficiency anemia. RESULTS: The mean age was 42.5 years. Lesions potentially causative of iron deficiency anemia were detected in 22 patients (20%). Upper gastrointestinal lesions were present in 14 patients (13%) and included only erosive lesions. Lower gastrointestinal lesions were detected in 8 patients (7.2%) and included colon cancer (2.7%), inflammatory bowel disease (3.6%), and a colonic ulcer >1 cm (0.9%). Patients with upper gastrointestinal lesions were more likely to use aspirin or nonsteroidal antiinflammatory drugs (11/14, 79%) than patients with no lesions (26/89, 23%; P = 0.043). Occult blood was more common in patients with lower gastrointestinal lesions 8/8 (100%) and patients with upper gastrointestinal lesions (9/14, 64%) than in patients without lesions (28/89, 31%; P = 0.037 and 0.039). Gastrointestinal symptoms were significantly more common in patients with gastrointestinal lesions than in patients without lesions. CONCLUSIONS: A gastrointestinal source of chronic blood loss was identified in a substantial proportion of premenopausal women with iron deficiency anemia. Patients with gastrointestinal symptoms, fecal occult blood, and/or weight loss should undergo endoscopy. PMID- 14745283 TI - High-dose versus low-dose clarithromycin in 1-week triple therapy, including rabeprazole and levofloxacin, for Helicobacter pylori eradication. AB - GOALS: To compare high-dose versus low-dose clarithromycin in 1-week triple therapy including rabeprazole and levofloxacin. BACKGROUND: Regimens containing rabeprazole and levofloxacin have proved to be effective against H. pylori infection. STUDY: One-hundred H. pylori-positive patients were randomly assigned to one of the following 1-week regimens: rabeprazole 20 mg o.d. plus levofloxacin 500 mg o.d. and clarithromycin 250 mg b.d. (RLC-1 group); rabeprazole 20 mg o.d. plus levofloxacin 500 mg o.d. and clarithromycin 500 mg b.d. (RLC-2 group). H. pylori status was assessed at entry and after the treatment. Patients who experienced treatment failure underwent antibiotic susceptibility testing. RESULTS: Forty-two patients in RLC-1 group (both PP and ITT analysis: 84%; 95%CI: 71-93%) and 47 in RLC2 group (both PP and ITT analysis: 94%; 95% CI: 83-98%) became H. pylori negative. Clarithromycin resistance was detected in all of 8 (100%) RLC-1 failures and in 1 out of 3 (33%) RLC-2 failures. Side effects occurred in 8% of patients in RLC-1 group and in 12% in RLC-2. CONCLUSIONS: Regimens tested are competitive with other PPI-based treatments. One-week triple therapy containing rabeprazole plus, levofloxacin, and high-dose clarithromycin yielded a higher eradicating rate than the one containing low-dose clarithromycin and may be considered as a first-line therapy option. PMID- 14745284 TI - Peripheral neuropathy as first sign of ulcerative colitis in a child. AB - A 6-year-old girl developed an axonal sensorimotor polyneuropathy a few days before intestinal symptoms with bloody diarrhea and abdominal pain. Colon biopsy established the diagnosis of ulcerative colitis. Vitamin and acid folic levels were normal. Peripheral neuropathy is an unusual event in ulcerative colitis in childhood, and an autoimmune pathogenesis is suspected. PMID- 14745285 TI - Medium-term results of endoscopic treatment of common bile duct strictures in chronic calcifying pancreatitis with increasing numbers of stents. AB - OBJECTIVES: The goal of this study was to evaluate our medium-term results on common bile duct stenting with increasing numbers of stents on strictures due to chronic calcifying pancreatitis. BACKGROUND: Common bile duct strictures frequently complicate the course of chronic calcifying pancreatitis. The effectiveness of endoscopic stenting to resolve definitely these strictures is still debated. STUDY: Twenty-nine patients with common bile duct stricture due to chronic calcifying pancreatitis were stented and followed up. Biliary sphincterotomy, dilation of the stricture, and insertion of plastic biliary stents (7.5-10 F) were performed. Patients were scheduled for elective stent changing/restenting at 3-month intervals or any time when it was urgently indicated. Our basic intention was to insert the maximum possible number of stents to reach as large diameter as the stricture allowed. All stents were removed after the disappearance of common bile duct dilatation or left in place in cases of persisting strictures. RESULTS: Eighteen patients (60%) had complete radiologic and serologic recovery after a mean of 21.1 months overall stenting time and had a stent free follow-up period for a mean of 12.1 months without recurrence of stricture. Five patients (16%) still have stents in place after 26 months. Three patients (13%) required surgery. There were 3 deaths (10%): 1 for unrelated cause and 2 with septic shock of biliary origin. CONCLUSIONS: Most chronic calcifying pancreatitis patients with common bile duct strictures respond to the increasing numbers of endoscopic stents, and remain stent free for medium term periods. Less patients (30%) does not benefit of biliary stenting, who are candidates for surgery. PMID- 14745286 TI - Which patients with chronic hepatitis B are more likely to relapse after interferon alpha-induced hepatitis B e antigen loss in Korea? AB - BACKGROUND AND AIMS: Although hepatitis B e antigen (HBeAg) loss after interferon alpha (IFN-alpha) therapy in patients with chronic hepatitis B is durable in whites, frequent relapse of hepatitis B virus (HBV) has been reported in endemic areas. This study was to evaluate the durability of interferon-induced HBeAg loss and find the risk factors for relapse. METHODS: We retrospectively analyzed 32 patients with IFN-alpha-induced HBeAg loss. They were followed up to 50 months. Age, sex, presumed route of infection, pretreatment levels of serum alanine aminotransferase and HBV DNA, and histologic findings were evaluated as potential risk factors for relapse. RESULTS: The cumulative relapse rates at 1-, 2- and 3 year were 25.5%, 35.7%, and 43.5%, respectively. They were higher in the older patients (>37 years) than in the younger ones (43.5% vs. 6.7% at 1-year, 50.6% vs. 20% at 2-year, and 64.7% vs. 20% at 3-year; P = 0.017). Patients with presumed vertical HBV infection tended to have a higher relapse rate (P = 0.11). Old age and presumed vertical infection were 2 independent risk factors for relapse. CONCLUSION: Relapse after IFN-alpha-induced HBeAg loss was common in this endemic area, especially in old patients and in those with presumed vertical HBV infection. Intensive surveillance and other therapeutic modalities would be necessary for these patients. PMID- 14745287 TI - Plasma and liver carnitine levels of children with chronic hepatitis B. AB - BACKGROUND: Carnitine status of children with chronic hepatitis B is not yet clear. Because it is well-known that carnitine interferes with T-cell immunity, which is closely related to spontaneous or treatment-induced seroconversion in hepatitis B, it was hypothesized that carnitine status would be effective on the end of therapy response (ETR). The aim of this study is to investigate both carnitine status of children with chronic HBV infection and its probable effects on liver histology and ETR. METHODS: Thirty-one children with chronic HBV infection, and age and sex matched 20 healthy children were included in the study. Plasma and liver free carnitine level determination was performed before IFN-alpha and lamivudine combination therapy in children with chronic hepatitis B. Mean plasma carnitine level of healthy children was used as control. IFN-alpha was injected as 5 million U/m2 subcutaneously 3 times a week for 6 months and lamivudine 4 mg/kg per day orally, maximum of 100 mg, for 1 year. RESULTS: The mean plasma carnitine level of patients with chronic HBV infection was significantly lower than that of controls (P < 0.001) The ETR was achieved in 14 (45.2%) patients. While plasma carnitine level was inversely correlated with portal inflammation score (P < 0.05), liver carnitine level was inversely correlated with fibrosis score (P < 0.05). CONCLUSION: It was found that plasma carnitine level was lower in children with chronic hepatitis B compared with healthy ones and carnitine was inversely correlated with liver portal inflammation and fibrosis scores. The role of carnitine in immunopathogenesis and histology of HBV needs to be clarified with further studies. PMID- 14745289 TI - Introducing fractals in magnification endoscopy. PMID- 14745288 TI - Lamivudine-induced acute pancreatitis in a patient with decompensated Hbv-related chronic liver disease. PMID- 14745290 TI - Hypoglycemic coma in a pregnant woman in association with hepatitis B virus carrier state and hepatocellular carcinoma. PMID- 14745291 TI - Spinal metastasis by hepatocellular carcinoma. PMID- 14745292 TI - Migrating shell fragment as a cause of recurrent obscure gastrointestinal bleeding. PMID- 14745295 TI - Are sobriety and consciousness determined by water in protein cavities? PMID- 14745296 TI - Association of GABRG3 with alcohol dependence. AB - BACKGROUND: Evidence from human, animal, and in vitro cell models suggests that gamma-aminobutyric acid (GABA), the major inhibitory neurotransmitter in the human central nervous system, is involved in many of the neurochemical pathways that affect alcohol use, abuse, and dependence. Both linkage and association to the region on chromosome 15q that contains a cluster of GABAA receptor genes have previously been reported, but the role of these genes in alcoholism remains inconclusive. METHODS: We conducted family-based association analyses by using a large sample of multiplex alcoholic families collected as part of the Collaborative Study on the Genetics of Alcoholism, to test for an association between alcohol dependence and the GABAA receptor genes clustered on chromosome 15q. Multiple single-nucleotide polymorphisms were tested in each of the three chromosome 15q GABAA receptor genes: GABRA5, GABRB3, and GABRG3. RESULTS: Using both classic trio-based analyses and extended-family analyses, we found consistent evidence of association between alcohol dependence and GABRG3. Nearly all single-nucleotide polymorphisms across the gene yielded evidence of association, and haplotype analyses were highly significant. No consistent evidence of association was observed with either GABRA5 or GABRB3, nor was there evidence for parent-of-origin effects with any of the genes. CONCLUSIONS: These analyses suggest that GABRG3 may be involved in the risk for alcohol dependence. These findings support the theory that the predisposition to alcoholism may be inherited as a general state of central nervous system disinhibition/hyperexcitability that results from an altered responsiveness to GABA. PMID- 14745297 TI - Alcohol dehydrogenase polymorphisms influence alcohol-elimination rates in a male Jewish population. AB - BACKGROUND: Genetic variation in the alcohol dehydrogenase (ADH) enzyme is associated with an aversion to alcohol and a lower risk of alcoholism among Asians. There is growing evidence of a functional role of the ADH2*2 allele in alcohol-drinking patterns among Jews, who have traditionally exhibited low rates of alcoholism and alcohol-related problems. The mechanism by which this allelic effect is mediated is not yet clearly understood. This study examined the effect of ADH2*2 on alcohol-elimination rates (AER) under experimental conditions. METHODS: Young adult male Jews (N = 109) received an intravenous alcohol infusion; metabolism was measured by using standard breath alcohol concentration tests. A clamping technique was used to achieve and maintain a target breath alcohol concentration of 50 mg/100 ml for a defined time period. The AER at steady state was calculated. The alcohol disappearance rate was also calculated from the descending limb slope. Polymerase chain reaction was used for allelic determination of the ADH2 and ADH3 loci. RESULTS: The mean AER among ADH2*2 carriers was significantly higher (8.09 +/- 1.4 g/hr) than among ADH2*1 homozygotes (7.14 +/- 1.5 g/hr; p = 0.003). Significance was retained on adjustment for potential confounding covariates. The ADH2 allele explains 8.5% of the AER variance in this population. Little AER difference was observed across ADH3 genotype groups. The slope of the descending limb increased with increasing copies of the ADH2*2 allele. CONCLUSIONS: The rate of alcohol elimination is significantly associated with the ADH2 genotype of Jewish males. Evidence for variation in alcohol metabolism across ADH genotypic groups provides support for the role of physiologic protective factors in alcohol drinking and suggests that reduced drinking among Jews may be genetically as well as environmentally determined. We believe that application of the novel "Indiana clamp" enhances AER measurement accuracy, allowing for detection of hitherto undetectable differences. PMID- 14745298 TI - Endogenous opioid receptor genes and alcohol dependence among Taiwanese Han. AB - BACKGROUND: Nonselective opioid antagonists reduce alcohol consumption under various experimental situations, and several association studies have examined possible roles of opioid receptor mu (OPRM), delta (OPRD), and kappa (OPRK) genes in the development of alcohol dependence. METHODS: We examined 20 single nucleotide polymorphisms (SNPs) across the OPRM, OPRD, and OPRK genes in 158 alcohol-dependent subjects and 149 controls. Differences in allele frequency and genotype distribution between case subjects and controls, as well as the deviation from Hardy-Weinberg equilibrium, were examined using Fisher's exact tests. RESULTS: No significant difference in either allele or genotype frequency was found between case subjects and controls for each of the SNPs. CONCLUSIONS: Our findings do not support a possible role of the opioid receptor genes for the proclivity to alcohol dependence in the Taiwanese Han. PMID- 14745299 TI - Determination of an estradiol dose-response relationship in the modulation of ethanol intake. AB - BACKGROUND: Estradiol (E2) potentiates the self-administration of numerous psychoactive drugs in female rodents. An analogous modulatory role of E2 on ethanol consumption remains unresolved because of examination of limited doses. The purpose of this study was to delineate a dose-response relationship for E2 on ethanol intake with an extended range and number of E2 doses. METHODS: Female Long-Evans rats had continuous access (22 hr/day) to both 10% ethanol (10E) solution and water. After the establishment of stable 10E intake baselines, rats were assigned to one of seven dose groups balanced for 10E intake [sham-operated (Shm) or ovariectomized (Ovx) plus E2 (microgram/kg)]: Shm + 0, Ovx + 0, Ovx + 0.05, Ovx + 0.15, Ovx + 0.5, Ovx + 1.5, and Ovx + 5. Ethanol preference drinking was evaluated during 25 consecutive days of E2 replacement treatment, and trunk blood was collected for the determination of plasma E2 and progesterone concentrations. RESULTS: Chronic E2 replacement regimens (0.05-1.5 micrograms/kg) dose-dependently augmented 10E intakes and ethanol preference ratios without concomitantly altering water consumption. Despite the maintenance of 2- to 3-fold greater plasma E2 levels, a supraphysiologic E2 dose (5 micrograms/kg) failed to precipitate ethanol intakes in excess of levels observed after treatment with a high physiologic E2 dose (1.5 micrograms/kg). Plasma progesterone concentrations were significantly increased in treatment groups (1.5 and 5 micrograms/kg E2) that exhibited corresponding significant increases in ethanol consumption. CONCLUSIONS: A positive dose-response relationship between E2 and ethanol intake (incremental increases in E2 dose corresponded to incremental increases in intake) was apparently limited to a physiologic concentration range, because a supraphysiologic dose failed to elicit an additional stepwise increase in ethanol intake. These findings stipulate a modulatory role for E2 in the regulation of moderate ethanol intake and suggest that endogenous fluctuations of E2 may alter the propensity toward consumption in women and in female animal models of ethanol self-administration. PMID- 14745300 TI - Is risk for alcoholism mediated by individual differences in drinking motivations? AB - BACKGROUND: Individual differences in motivations to drink have been proposed as a mechanism that mediates risk for alcoholism. We investigated the genetic and environmental sources of variation in motivations for drinking, as assessed by four scales of the Alcohol Use Inventory (AUI), and then examined the extent to which genetic and environmental variations in risk for alcoholism are mediated by individual differences in drinking motives. METHODS: Data on four AUI scales (assessing drinking to manage mood states, to relieve social anxiety, in social situations, and to improve mental functioning) and lifetime DSM-IV alcohol abuse and/or dependence (AAD) were obtained from 2529 female and 3709 male adult twins, including 2229 complete twin pairs, from the population-based Virginia Twin Registry. RESULTS: Logistic regression analyses indicated that higher scores on each of the four AUI variables were significantly associated with AAD, with increases in risk for diagnosis of 40% to 141% per standard deviation increase in AUI score. Structural modeling analyses conducted using Mplus indicated that individual differences in AUI scores were in part due to genetic variation, particularly among women. Among males, genetic factors were substantial for drinking to alter mood but small for other measures. A substantial portion of the genetic variation in AAD overlapped with drinking to manage mood states. Results from bivariate twin models of AAD and the AUI scales were consistent with the mediation hypothesis for the social anxiety and social interaction scales but not drinking to manage mood or to enhance mental functioning. CONCLUSIONS: Genetic contributions to variation in risk for alcoholism may be mediated in part by individual differences in motivations related to drinking in social settings. Drinking to manage mood indexes genetic risk for alcoholism but does not appear to act as a direct cause of alcoholism. PMID- 14745301 TI - Acute ethanol withdrawal (hangover) and social behavior in adolescent and adult male and female Sprague-Dawley rats. AB - BACKGROUND: The extensive use of alcohol during adolescence may be facilitated by an age-specific attenuation in sensitivity to adverse effects of ethanol. Adolescent rats are less sensitive than adults to some delayed effects of acute ethanol, including hangover-related anxiety on an elevated plus maze. The purpose of this study was to determine whether adolescent rats are also less sensitive than adults to hangover-related anxiogenesis when indexed in terms of social inhibition. METHODS: Anxiety during ethanol hangover was indexed in adolescent and adult Sprague-Dawley rats of both genders by assessing the suppression in social behavior during a social interaction test. Animals were tested 18 hr after intraperitoneal administration of 0 g/kg (saline) or 4 g/kg ethanol (experiment 1) or at test intervals chosen on the basis of assessments of ethanol clearance time (experiment 2) to equate clearance-to-test intervals across age and gender (experiments 3-5). RESULTS: Adults showed more hangover-related social suppression and slower postclearance recovery than adolescents. Sex differences were more pronounced in adults than adolescents, with males being more affected than females. Ethanol clearance time was considerably longer in adult males than in adolescent animals and adult females. In contrast to the modest decreases in social activity observed in adolescent animals shortly after ethanol clearance, adolescents showed a surprising increase in play fighting later in the recovery period- a hangover-related social facilitation that was not evident in adults. CONCLUSIONS: The attenuated anxiety and increase in social interactions seen in adolescents during hangover are less likely to serve as deterrents for further drinking than the aversive increase in anxiety seen in adults. A facilitation of social interactions not only during a drinking episode, but also during the postalcohol recovery period, could help to establish a persisting cycle of drinking in at-risk adolescents, leading to dependency and a lifelong history of alcohol-related problems. PMID- 14745302 TI - The efficacy of acamprosate in the maintenance of abstinence in alcohol-dependent individuals: results of a meta-analysis. AB - BACKGROUND: A number of clinical trials have been undertaken to determine the efficacy of acamprosate in the maintenance of abstinence in alcohol-dependent individuals. However, the reported differences in patient populations, treatment duration, and study endpoints make comparisons difficult. An assessment of the efficacy of treatment with acamprosate was, therefore, undertaken using meta analytical techniques. METHODS: All randomized, placebo-controlled trials (RCTs) that fulfilled predetermined criteria were identified using (1) a language unrestricted search of 10 electronic databases; (2) a manual search of relevant journals, symposia, and conference proceedings; (3) cross-referencing of all identified publications; (4) personal communications with investigators; and (5) scrutiny of Merck-Sante's internal reports of all European trials. Study quality was assessed, independently, by three blinded workers. Key outcome data were identified; some outcome variables were recalculated to ensure consistency across trials. The primary outcome measure was continuous abstinence at 6 months; abstinence rates were determined by estimating Relative Benefit (RB). RESULTS: A total of 19 published 1 unpublished RCTs were identified that fulfilled the selection criteria; 3 were excluded because the documentation available was insufficient to allow adequate assessment. The remaining 17 studies, which included 4087 individuals, 53% of whom received active drug, were of good quality and were otherwise reasonably comparable. There was no evidence of publication bias. Continuous abstinence rates at 6 months were significantly higher in the acamprosate-treated patients (acamprosate, 36.1%; placebo, 23.4%; RB, 1.47; [95% confidence intervals (CI): 1.29-1.69]; p < 0.001). This effect was observed independently of the method used for assigning missing data. The effect sizes in abstinent rates at 3, 6, and 12 months were 1.33, 1.50, and 1.95, respectively. At 12 months, the overall pooled difference in success rates between acamprosate and placebo was 13.3% (95% CI, 7.8-18.7%; number needed to treat, 7.5). Acamprosate also had a modest but significant beneficial effect on retention (6.01%; [95% CI, 2.90-8.82]; p = 0.0106). CONCLUSION: Acamprosate has a significant beneficial effect in enhancing abstinence in recently detoxified, alcohol-dependent individuals. PMID- 14745303 TI - Improvement in quality of life after treatment for alcohol dependence with acamprosate and psychosocial support. AB - BACKGROUND: The impact of disease on health-related quality of life is now well recognized, as is the importance of this variable as a measure of treatment efficacy. METHODS: Patients from five European countries were enrolled in an open, multicenter, prospective study designed to observe outcome in dependent drinkers treated for 6 months with acamprosate and psychosocial support. Version 1 of the 36-item Short Form Health Profile (SF-36v1) questionnaire was administered at inclusion and at 3 and 6 months. Responses were described as handicaps compared with an appropriately matched, healthy reference population. One-way fixed ANOVA and simultaneous stepwise linear regression analysis were used to identify potential predictors of quality of life at baseline and after treatment. RESULTS: Baseline SF-36v1 data were obtained from 1216 patients (mean age, 43 +/- 9 years; 77% male). Mean values for all SF-36v1 dimensions were significantly lower in the patient population than in the normative reference population; the most important deficits were observed in physical and emotional role limitations and in social functioning. The most important predictors of baseline quality of life were severity of alcohol dependence, employment status, psychiatric history, quantity and frequency of alcohol consumption, attendance at Alcoholics Anonymous, global alcohol health status, age, gender, and education. SF-36v1 data were obtained from 686 patients at 3 months and from 497 at 6 months. Significant improvements were observed in all SF-36v1 dimensional and summary scores after 3 months of treatment (p < 0.001); further marginal improvements were observed between 3 and 6 months. The most important predictors of quality of life following treatment were the SF-36v1 profile at baseline, followed by abstinence duration; patients who completed the trial and remained abstinent throughout showed the greatest improvement. CONCLUSIONS: Health-related quality of life is severly impaired in dependent drinkers. Treatment with acamprosate and psychosocial support, by promoting abstinence, improves the quality of life profile to levels comparable to those observed in healthy individuals. PMID- 14745304 TI - Effective strategies for recruiting families ascertained through alcoholic probands. AB - BACKGROUND: Recruiting a large number of participants meeting strict inclusion criteria can be challenging, particularly when selecting for a condition associated with a social stigma such as alcoholism, when participation involves collection of medical specimens and sensitive information, and when the participation of family members or other collaterals is required. Developing and implementing a successful recruitment plan depends upon identifying the most effective recruitment strategies given the available resources. METHODS: Several strategies for recruiting subjects for a large family study on the genetics of alcoholism were evaluated over a two-year period with regard to participant yield, time and cost expenditure. RESULTS: Overall effectiveness of a recruitment strategy was determined based on a composite of yield, cost and time expenditure. The most effective recruitment strategies were direct mail, press release, the Internet and treatment center collaborations. CONCLUSION: Results provide insight into successful strategies for recruiting large numbers of participants and their family members selected for a condition associated with a social stigma. PMID- 14745305 TI - Ethanol withdrawal influences survival and morphology of developing rat hippocampal neurons in vitro. AB - BACKGROUND: Previous studies in this laboratory have shown that, like their counterparts in vivo, fetal rat hippocampal pyramidal neurons in culture develop abnormally small dendritic arbors when exposed to ethanol. This study asked whether ethanol's inhibitory effects on dendritic development differ when the duration of ethanol exposure and timing of withdrawal are varied to correspond with early versus later stages of development and whether ethanol withdrawal influences survival of these neurons. METHODS: We compared neurons exposed continuously for 6 or 14 days to ethanol (70 mM) with neurons transferred from ethanol-containing medium to control medium either 1 day after adding ethanol (before dendrites elongated) or 6 days after adding ethanol (after dendrites began elongating). We then performed morphometric and cell density analyses at 6 and 14 days using digital images of neurons immunostained with microtubule associated protein 2 (MAP2) to visualize dendrites. RESULTS: Continuous exposure to ethanol decreased the length and number of dendrites formed but had no effect on neuron survival compared with controls without ethanol. Dendritic length was less inhibited when ethanol was withdrawn after 1 day, but the number of dendrites per cell was unchanged compared with neurons continuously exposed to ethanol. Withdrawal from ethanol at 1 day slightly enhanced the survival of neurons assessed at 14 days compared with neurons in control medium and with neurons exposed continuously to ethanol. In contrast, withdrawal from ethanol at 6 days severely decreased the number of neurons at 14 days. CONCLUSIONS: These results suggest that dendrites can achieve normal length when ethanol exposure is limited to only 1 day and withdrawal occurs before dendrites begin elongating. However, a persistent reduction in dendrite number results in smaller overall dendritic arbor size. Although continuous exposure to ethanol has little effect on neuron survival in these cultures, and exposure limited to 1 day followed by withdrawal can be neuroprotective against cell death associated with increased time in culture, longer exposure before withdrawal can trigger cell death. PMID- 14745306 TI - Survival of AA and ANA rats during lifelong ethanol exposure. AB - BACKGROUND: Study of the long-term effects of chronic alcohol consumption in human populations is confounded by genetic and environmental factors. METHODS: The study was intended to investigate the effects on morbidity and survival of lifetime forced ethanol consumption in male and female AA (Alko, Alcohol) and ANA (Alko, Non-Alcohol) rats. The ethanol-exposed rats had 12% ethanol as the only available fluid from 3 to 24 months of age. The control groups had water. Rats that died during the experiment and those that were killed at 24 months of age were all autopsied, and the pathologic findings were recorded. RESULTS: Lifelong ethanol consumption did not change the survival rate of the rats, and had no significant effect on the rates of any of the pathologic measures in either the AA or ANA line of rats, suggesting that this may not be a good animal model for studying the detrimental effects of chronic alcohol. An unexpected, highly significant finding was observed: the AA rats, bred for high voluntary ethanol drinking, lived much longer than the ANA rats, bred for ethanol avoidance. The death rate by 24 months in the AA line was less than one-third of that in the ANA line. This difference was found regardless of whether the animals were maintained on alcohol or water, and in both genders. The AA rats had significantly lower rates of kidney disease, benign tumors, and cardiovascular disease than the ANA animals. CONCLUSIONS: Lifelong ethanol consumption increased neither the mortality nor the morbidity of AA and ANA line of rats. Genes selected in the development of the high drinking AA line have additional effects producing rats that are healthier and living longer than the ANA rats possessing genes resulting in alcohol avoidance. PMID- 14745307 TI - Chronic ethanol consumption reduces delta-and mu-opioid receptor-stimulated G protein coupling in rat brain. AB - BACKGROUND: Ethanol consumption is thought to enhance the release of endogenous opioids acting at opioid receptors (ORs) in the central nervous system. Prior studies have shown that chronic ethanol consumption in alcohol-preferring rats uncouples mu-ORs from Gi proteins. The purpose of this study was to investigate the potential for uncoupling of the delta- and the mu-OR after chronic ethanol consumption in a nonpreferring rat strain. METHODS: We used radiohistochemical methods to study mu- and delta-OR-stimulated G-protein coupling in brain tissue of rats ingesting liquid diets containing 6.7% ethanol (v/v) for 16 days, as compared with 0% ethanol pair-fed control rats. Sections of brain from pair-fed and ethanol-treated rats were incubated with guanylyl 5'-[gamma-[35S]-thio] triphosphate ([35S]-GTPgammaS) in the absence and presence of d-Pen2,d-Pen5 enkephalin (DPDPE), a delta-OR agonist, or Tyr-d-Ala-Gly-N(me)Phe-Gly-ol enkephalin (DAMGO), a mu-OR agonist. RESULTS: DPDPE significantly stimulated [35S]-GTPgammaS binding in the hippocampal dentate gyrus (DG), CA1, cerebellum, and inferior colliculus of untreated pair-fed controls. By contrast, DPDPE stimulated [35S]-GTPgammaS binding was reduced significantly in those brain regions in the ethanol-consuming group. DAMGO stimulated [35S]-GTPgammaS binding in cortex, caudate, nucleus accumbens, DG, CA1, and superior and inferior colliculi, whereas the DG, CA1, and colliculi showed a significant reduction of binding after chronic ethanol. Basal [35S]-GTPgammaS binding was not different between the two diet groups. CONCLUSIONS These data are the first to demonstrate functional uncoupling of delta-ORs from G proteins after chronic ethanol consumption. Uncoupling may result from modulation of receptors, possibly by internalization or phosphorylation. Alterations in functional coupling of both delta- and mu-ORs and subsequent effects may contribute to continued ethanol consumption. PMID- 14745308 TI - NMDA receptor subunit expression after combined prenatal and postnatal exposure to ethanol. AB - BACKGROUND: The N-methyl-D-aspartate receptor (NMDAR), a subtype of glutamate receptor, is essential for normal neurodevelopment. The brain growth spurt, which is both prenatal and postnatal in the rat, is a time when the brain is especially sensitive to the effects of a teratogen, such as alcohol. Changes in NMDAR function after early perinatal exposure to ethanol (EtOH) may be related to alterations in the expression of secondary subunits. Thus, we investigated the expression of the NR1, NR2A, and NR2B subunits after combined prenatal and postnatal exposure to EtOH. METHODS: A binge model was used to administer EtOH (5 g/kg) or isocaloric vehicle to pregnant female rats followed by EtOH (6.2 g/kg) or isocaloric control diet from postnatal days 4 through 9 via an artificial rearing method. Proteins from crude membrane homogenates isolated from cortex and hippocampus at postnatal day 10, 14, or 21 were separated in a standard Western blot procedure. RESULTS: The expression of the NR2A subunit of EtOH-exposed pups showed a significant increase at postnatal day 10 in hippocampus compared with diet controls. No significant changes were seen for any other subunit in either region. CONCLUSIONS: The up-regulation of NR2A during EtOH withdrawal is consistent with compensatory changes to prolonged inhibition of the NMDAR. These results indicate that postnatal exposure to ethanol produces distinct effects on the NMDAR, which may underlie deficits associated with alcohol-related neurodevelopmental disorder. PMID- 14745309 TI - Fetal alcohol exposure alters GAP-43 phosphorylation and protein kinase C responses to contextual fear conditioning in the hippocampus of adult rat offspring. AB - BACKGROUND: The growth- and plasticity-associated protein GAP-43 plays a significant role in the establishment and remodeling of neuronal connections. We have previously shown that GAP-43 levels, protein kinase C (PKC) activity, and GAP-43 phosphorylation increase during contextual fear conditioning and that fetal alcohol exposure (FAE) decreases PKC activity and GAP-43 phosphorylation in the hippocampus of adult offspring. Drawing on these observations, we hypothesized that FAE manifests its cognitive impairment by disrupting PKC activation and membrane translocation, thereby decreasing GAP-43 phosphorylation and function. METHODS: Three groups of pregnant rat dams (FAE and two control diet groups) were placed on different diet regimens. Offspring from each of these groups were placed into each of four test groups, a contextual fear conditioned (CFC) group, a naive unhandled group, and two nonlearning stress control groups. Hippocampi were dissected, homogenized, and used to prepare a cytosolic and a membrane fraction. These fractions were probed for total GAP-43, PKC phosphorylated GAP-43, and several PKC subtypes. PKC activity also was measured in total homogenates. RESULTS: Compared with both control diet groups, FAE animals showed a deficit in the activation of PKC in the hippocampus at 24 hr but not at 1.5 hr after CFC. Likewise, we found that the amount of GAP-43 and its phosphorylation were decreased 24 hr after CFC in FAE rats but not at early times after training. Analysis of the translocation of various PKC isoforms revealed that FAE animals had decreased levels of membrane-bound PKC beta2 and PKC epsilon 24 hr after CFC. CONCLUSIONS: Considering the role of PKC activation and GAP-43 phosphorylation in synaptic plasticity, our results suggest that deficient translocation of PKC beta2 and PKC epsilon in the hippocampus may mediate the electrophysiological and behavioral deficits observed in fetal alcohol exposed animals. PMID- 14745310 TI - Postnatal binge ethanol exposure affects habituation of the cardiac orienting response to an olfactory stimulus in preweanling rats. AB - BACKGROUND: Prenatal exposure to alcohol can result in intellectual impairments, reduced habituation and distractibility, visual-spatial deficits, and problems in attention. Dysfunction in attention, including habituation of responses to nonsignal stimuli, can have devastating consequences for cognitive development. This research examined whether binge exposure to ethanol on postnatal days (PD) 4 to 9 would affect cardiac orienting or response habituation to an olfactory stimulus in preweanling rats. METHODS: Ethanol-exposed subjects were given ethanol 5.25 g/kg/day on PD4 to 9. Controls were either sham intubated or unhandled during this time. To assess baseline and phasic cardiac function, PD16 subjects were tested under differential pharmacological blockade of the sympathetic or parasympathetic nervous systems by administration of the peripherally acting antagonists atenolol or atropine methyl nitrate. In a second experiment, separate groups of subjects were tested for habituation of the cardiac orienting response to an olfactory stimulus. RESULTS: Results indicate that postnatal ethanol had no observable effect on the functional development of autonomic control over heart rate. Similarly, ethanol exposure had no effect on the form or magnitude of the cardiac orienting response. However, neonatal ethanol exposure did result in a deficit in orienting response habituation; ethanol-exposed subjects continued to respond to the stimulus with a large magnitude bradycardia after control subjects exhibited complete response habituation. In addition, ethanol-exposed subjects had longer orienting response latencies than controls. CONCLUSIONS: The results indicate that this animal model has the potential to contribute to investigations designed to understand basic forms of memory impairments observed in humans with a history of prenatal alcohol exposure. The postnatal binge ethanol model of fetal alcohol effects produces deficits in nonassociative memory that are similar to those observed in human infants exposed prenatally to ethanol. Deficits in response habituation have important implications for cognitive development. PMID- 14745311 TI - Liver disease in heavy drinkers with and without alcohol withdrawal syndrome. AB - BACKGROUND: Withdrawal syndrome is a hallmark of alcohol dependence. The characteristics of alcohol consumption, closely related to dependence, could influence the development of alcoholic liver disease. The study aimed to investigate if patients with severe alcohol withdrawal syndrome have a peculiar profile of liver disease. METHODS: The study included 256 heavy drinkers (aged 19 75 years, 70.3% males) admitted to an Internal Medicine Department. Patients admitted for complications of liver disease were not included. Severe alcohol withdrawal syndrome (seizures, disordered perceptions, or delirium) developed in 150 patients (58.6%). Alcohol consumption (daily quantity, duration, and pattern [regular or irregular]) was assessed by questionnaire. Liver biopsy was performed in all cases. RESULTS: Patients with alcohol withdrawal syndrome showed a lower prevalence of liver cirrhosis and a higher prevalence of alcoholic hepatitis than patients without it. The negative association of alcohol withdrawal syndrome with liver cirrhosis persisted after we adjusted for sex, daily intake, duration, and pattern of alcohol consumption. Alcoholic hepatitis was independently associated with the irregular pattern of alcohol consumption, which was closely associated with severe alcohol withdrawal syndrome. CONCLUSIONS: The profile of liver injury is different in heavy drinkers who develop and who do not develop a severe alcohol withdrawal syndrome when admitted to the hospital. PMID- 14745312 TI - Mismatch negativity: no difference between controls and abstinent alcoholics. AB - BACKGROUND: A number of studies have examined the amplitude of the mismatch negativity (MMN) evoked potential as a measure of a brain inhibitory deficit in alcoholics or those at risk for alcoholism. The current study examined MMN in alcoholics abstinent an average of 6.7 years (with a minimum of six months abstinence) compared to controls. This study examined the association of MMN with alcoholism family history density, with indices of the presence and severity of externalizing disorders (a risk-factor for alcoholism), and with alcohol use variables. METHODS: Electroencephalograms were gathered on 76 subjects (38 controls, 38 abstinent alcoholics) during a nonattending mismatch negativity experiment. Measures of alcoholism family history density, disinhibited personality traits, and antisocial symptoms served as measures of risk-factors known to be associated with a genetic liability to alcoholism. Alcohol use variables were used as measures of alcoholism severity. RESULTS: There were no differences in MMN amplitude or latency between controls and abstinent alcoholics. There also were no significant associations between MMN measures and the measures of risk for alcoholism or with the severity of alcohol use or duration of abstinence. CONCLUSIONS: The results suggest that MMN is neither affected in chronic alcoholics nor associated with alcoholism vulnerability, and thus does not reflect a trait marker of alcoholism or alcoholism risk. The current results do not address effects on MMN of acute alcohol ingestion or withdrawal from alcohol. PMID- 14745313 TI - Ethanol selectively inhibits enhanced vesicular release at excitatory synapses: real-time visualization in intact hippocampal slices. AB - BACKGROUND: Conflicting information exists concerning the actions of ethanol on vesicular release at excitatory synapses. Because long-term alterations in synaptic transmission are thought to underlie neuroadaptive responses to ethanol, we have directly measured the actions of ethanol on release dynamics at an intact central synapse. METHODS: Here we investigated the effects of ethanol on release dynamics in hippocampal slices using confocal microscopy with the lipophilic dye, FM1-43, complemented by a patch clamp analysis of AMPA miniature excitatory postsynaptic currents (mEPSCs). After a pretreatment/loading paradigm with sulforhodamine (S-Rhd) and FM1-43, stable, dense punctate FM1-43 staining in the CA1 stratum radiatum was evident. RESULTS: FM1-43 fluorescence destaining was dose-dependently induced by perfusion with elevated K+ (20-60 mM). Cadmium inhibited K+-induced destaining, whereas nifedipine had no significant effect. Ethanol (25-75 mM) inhibited K+-induced destaining with high efficacy and had no effect on basal destaining. Both omega-Conotoxin GVIA and omega-Agatoxin IVA inhibited K+-induced destaining with high efficacy. The combination of omega Conotoxin GVIA and omega-Agatoxin IVA occluded the inhibitory effect of ethanol, indicating that ethanol inhibition of release was dependent on inhibition of N/P/Q-voltage-gated calcium channels (VGCCs). Patch clamp studies of AMPA mEPSCs revealed similar findings in that vesicular release was enhanced with K+ depolarization in an ethanol-sensitive manner. CONCLUSIONS: These findings indicate that the FM1-43/S-Rhd method is a stable and powerful approach for direct real-time measurement of vesicular release kinetics in intact brain slice preparations and that ethanol inhibits vesicular release induced by depolarization via inhibition of N/P/Q-VGCCs. PMID- 14745314 TI - Ethanol interferes with the measurement of extracellular ionized calcium. AB - BACKGROUND: Maintenance of extracellular calcium (Ca) concentration within narrow limits is critical for normal cell function and optimization of bone health. Ionized Ca (iCa), the form of Ca that is regulated, has been reported to vary inversely with blood alcohol concentration (BAC). The mechanism by which ethanol decreases blood iCa levels is unknown. However, one possible explanation is that it is, at least partially, a methodological artifact due to an effect of ethanol on the function of the ion selective electrode used to measure iCa. The purpose of this study was to determine if ethanol interferes with the measurement of iCa and if this interference can account for the typically observed in vivo effects of ethanol consumption on blood iCa levels. METHODS: Ethanol (0-5 microl/ml) was added to blood or iCa standards and the iCa concentration measured using the ICA2 iCa analyzer (Radiometer) or the I-Stat Clinical Analyzer (Abbott Laboratories). Both instruments use an ion selective electrode to measure iCa. The relationship between iCa and BAC determined from the ex vivo addition of ethanol to blood was compared with relationships obtained in vivo during chronic ethanol consumption. RESULTS: Addition of ethanol to blood or iCa standards resulted in a dose dependent decrease in iCa concentration when iCa was measured using the ICA2 analyzer but had no effect on iCa concentration when measured with the I-Stat Clinical Analyzer. Ethanol's effect on iCa with the ICA2 analyzer did not differ between blood and standards, and ethanol had no effect on pH, suggesting that the ethanol-induced decrease in iCa was due to a methodological artifact. However, the magnitude of ethanol's effect was small and does not account entirely for the relationship between iCa and BAC observed in vivo with chronic ethanol consumption. CONCLUSION: Ethanol can interfere with the measurement of iCa using ion selective electrodes, but this interference depends on the analyzer used. This is a significant methodological issue that has largely been unrecognized in the field of alcohol research. Although this interference does not explain entirely the relationship between iCa and BAC observed in vivo in studies on chronic ethanol consumption, it complicates investigations designed to assess the effect of ethanol on Ca homeostasis. PMID- 14745315 TI - Alcohol, but not lipopolysaccharide-induced liver apoptosis involves changes in intracellular compartmentalization of apoptotic regulators. AB - BACKGROUND: While alcohol-induced augmentation of liver apoptosis has been demonstrated in humans and laboratory animals, the underlying mechanisms are not fully elucidated. This study addresses the question whether alcohol and bacterial lipopolysaccharide (LPS), a putative mediator of alcohol effects on the liver, induce augmentation of liver apoptosis by intrinsic or extrinsic signaling pathways. This information may prove important for future design of therapies for alcoholic liver disease. METHODS: Male rats were fed either an alcohol-containing liquid diet or an isocaloric, control diet for 15-16 weeks. At the end of feeding period, the rats were treated with LPS (0.8 mg.kg-1 body weight) or sterile saline and killed 3 and 24 hr later. The liver and blood were sampled for histology and biochemical assays. Hepatocytes were isolated by collagenase perfusion and fractionated to yield mitochondria and cytoplasm. The propensity of mitochondria to undergo permeability transition in the presence of a Ca2+ overload was determined along with distribution of various apoptotic regulators (AIF, Smac2, Bax, cytochrome c, Bcl-XL, Bfl-1, and caspase-2) between mitochondria and cytoplasmic fractions. RESULTS: Increased liver apoptosis in alcohol-treated rats was associated with translocation of several apoptotic regulators between mitochondria and cytoplasm in a manner suggesting that alcohol induces augmentation of apoptosis by recruiting intrinsic apoptotic signals. LPS treatment of rats counteracted alcohol-induced changes in intracellular compartmentalization of apoptotic regulators despite an increased rate of apoptosis. LPS may, therefore, recruit extrinsic apoptotic signals, such as proinflammatory cytokines. CONCLUSIONS: Hepatocytes are to be able to mount an apoptotic response to both intrinsic and extrinsic signals. Alcohol increases liver apoptosis predominantly through an intrinsic signaling pathway while LPS recruits extrinsic signaling pathways. PMID- 14745316 TI - Abnormal hepatic methionine and glutathione metabolism in patients with alcoholic hepatitis. AB - BACKGROUND: Abnormal methionine metabolism occurs in animals fed ethanol and in end-stage cirrhotic patients. Expected consequences of these abnormalities include reduced hepatic S-adenosylmethionine and glutathione (GSH) levels, impaired transmethylation, and reduced homocysteine catabolism, resulting in the often-observed hyperhomocystinemia in cirrhotic patients. These parameters have not been examined simultaneously in patients with less advanced alcoholic liver disease. METHODS: Six patients hospitalized for alcoholic hepatitis were studied. Plasma was analyzed for homocysteine, methionine, and GSH levels. Liver biopsies diagnosed acute alcoholic hepatitis and underlying fibrosis. Liver specimens were processed for messenger RNA (mRNA) levels and various metabolites and were compared with those of six normal controls. RESULTS: Three patients had cirrhosis, and three had only portal fibrosis. Plasma levels of homocysteine and methionine were increased in two of the three patients with cirrhosis but not in the patients with fibrosis. All patients had markedly lower plasma GSH levels (mean +/- SD: 0.27 +/- 0.19 microM, which is at least 10-fold lower than the normal range). Hepatic S-adenosylmethionine levels were reduced by 50%, whereas methionine, GSH, and cysteine levels were reduced by 70-80%. The mRNA levels of most enzymes involved in methionine metabolism and GSH synthesis were decreased, whereas albumin expression was unchanged. Despite the well known induction of cytochrome P450 2E1 in chronic alcoholics, its mRNA levels were nearly 70% lower in these patients. CONCLUSIONS: In alcoholic hepatitis, abnormal hepatic gene expression in methionine and GSH metabolism occurs and often contributes to decreased hepatic methionine, S-adenosylmethionine, cysteine, and GSH levels. It may be important to replenish these thiols in patients hospitalized with alcoholic hepatitis. PMID- 14745318 TI - Basic certification in surgery by the American Board of Surgery (ABS): what does it mean? Does it have value? Is it relevant? A personal opinion. PMID- 14745317 TI - The effects of binge alcohol exposure on bone resorption and biomechanical and structural properties are offset by concurrent bisphosphonate treatment. AB - BACKGROUND: Chronic alcohol consumption reduces bone mass and strength, increasing fracture risk for alcohol abusers. Mechanisms underlying this vulnerability involve modulation of bone remodeling. Direct effects of alcohol on bone formation have been documented; those on bone resorption are less well studied. Skeletal effects of exposure to high blood alcohol concentrations (BAC's) attained during binge drinking have not been studied. We examined the effects of repeated binge-like alcohol treatment on bone resorption, bone mineral density and vertebral compressive strength in adult male rats treated with the aminobisphosphonate, risedronate. METHODS: A binge alcohol exposure model was developed using intraperitoneal (IP) injection to administer a 20% (vol/vol) alcohol/saline solution (3 g/kg, 1X/day) on four consecutive days for 1, 2 or 3 weeks in 400 g rats, with and without weekly risedronate treatment (0.5 mg/kg, 1X/week). Total serum deoxypyridinoline (Dpd) a crosslink of bone type collagen released during resorption was measured by ELISA. Bone mineral density (BMD) was measured using peripheral quantitative computed tomography (pQCT). Vertebral compressive strength was determined using an Instron materials testing machine. Trabecular integrity was analyzed by computer-aided trabecular analysis system (TAS). RESULTS: Peak BAC's averaged 308.5 +/- 12 mg/dL; average BAC was 258.6 +/- 28.7 mg/dL at time of euthanasia. No significant effects of treatment were observed after 1 or 2 weeks of binge alcohol exposure. At 3 weeks of alcohol treatment serum Dpd was significantly increased (205%, p < 0.05) over controls. Bone mineral density (BMD) in cancellous bone of distal femur and lumbar spine were significantly decreased (34% and 21% respectively, p < 0.01) after 3 weeks of binge treatment. Vertebral (L4) compressive strength (maximum load sustained before failure) also decreased (27%, p < 0.05) after 3 binge alcohol cycles. Risedronate maintained the Dpd level (p < 0.01), BMD (p < 0.001) and vertebral structural biomechanical properties (p < 0.01) of binge-treated rats at control levels (E vs ER). Indices of trabecular architectural integrity [Trabecular bone volume/tissue volume (BV/TV), bone area (BAR) and trabecular separation (Tb.Sp)] analyzed at week 3 showed (BV/TV) and (BAR) were significantly reduced in alcohol binged rats (p < 0.01), while (Tb.Sp) was significantly increased (p < 0.01). Risedronate also maintained the trabecular architectural indices of binge-treated rats at control levels (E versus ER, p < 0.01). CONCLUSIONS: In adult male rats, BAC's reflective of those attained during alcoholic binge drinking may affect the skeleton in part by stimulating bone resorption, an effect mitigated by risedronate. PMID- 14745319 TI - Don't be afraid of change: a commentary on surgical training and the American Board of Surgery. PMID- 14745320 TI - Adult living donor liver transplantation for patients with hepatocellular carcinoma: extending UNOS priority criteria. AB - INTRODUCTION: For patients with hepatocellular carcinoma (HCC) in particular, living donor liver transplant (LDLT) improves access to transplant. We report our results in 36 patients with HCC who underwent LDLT with a median follow-up >1 year. METHODS Underlying diagnoses included: hepatitis C (24), hepatitis B (9), cryptogenic cirrhosis (1), hemochromatosis (1), and primary biliary cirrhosis (1). Patients with tumors >or= 5 cm received IV doxorubicin intraoperatively and 6 cycles of doxorubicin at 3-week intervals. Patients were followed with CT scan and alpha-fetoprotein levels every 3 months for 2 years posttransplant. Mean waiting time, pretransplant treatment, tumor variables, and survival were analyzed. Univariate and multivariate analysis were done to analyze tumor variables; Kaplan-Meier and log rank were used to compare survivals. P < 0.05 was considered significant. RESULTS Mean wait for LDLT was 62 days, compared with 459 days in 50 patients with HCC transplanted with cadaveric organs during the same time period (P = 0.0001). At median follow-up of 450 days, there have been 10 deaths due to non-tumor-related causes and 3 deaths from recurrence; recurrence has also been observed in 3 other patients. On univariate and multivariate analysis, bilobar distribution was the only significant tumor variable (P = 0.03, log rank = 0.02). Fifty-three percent of patients exceeded UNOS priority criteria. One- and two-year patient survivals were 75% and 60%, respectively. Freedom from recurrence at 365 and 730 days was 82% and 74%, respectively. Overall and in patients with HCC > 5 cm (n = 12), there were no statistically significant differences in survival or in freedom from recurrence between recipients of living donor and cadaveric grafts. CONCLUSION Although one third of patients had tumors > 5 cm, the incidence of recurrence as well as patient survival and freedom from recurrence are comparable to results after cadaveric transplant. LDLT allows timely transplantation in patients with early or with large HCC. PMID- 14745321 TI - Liver transplantation for the treatment of moderately or well-differentiated hepatocellular carcinoma. AB - OBJECTIVE: To determine the long-term results of liver transplantation for well- or moderately differentiated hepatocellular carcinoma (HCC). SUMMARY BACKGROUND DATA: HCC patient selection for liver transplantation remains controversial, and deciding exclusively on the strength of criteria such as number and size of nodules appears prognostically inaccurate. METHODS: Since 1991, preoperative tumor grading has been used at our center to establish whether a patient with HCC is fit for transplantation. Poorly differentiated HCC cases were excluded, while size and number of nodules were not considered as absolute selection criteria. Thirty-three patients with moderately or well-differentiated HCC were prospectively studied after liver transplantation. A group of 15 patients with incidental HCC transplanted during the same period were also evaluated and compared with the 33 patients with preoperatively diagnosed HCC. RESULTS: On histologic examination, 38% of the entire group of 48 patients did not meet the "Milan criteria" and 42% were pTNM stages III and IV. The median follow-up was 44 months. The 5-year actuarial survival rate was 75% and recurrence-free survival was 92%. HCC recurred in only 3 patients (6%). The only histomorphologic variable differing significantly between incidental and nonincidental HCC was nodule size. The timing of diagnosis (incidental vs. nonincidental HCC), the Milan criteria, and the TNM stage revealed no statistically significant impact on overall and recurrence-free survival rates. CONCLUSIONS: The routine pre-orthotopic liver transplantation tumor grading may represent a valid tool in the selection of unresectable HCC patients for transplantation. PMID- 14745322 TI - Challenging choices: liver transplantation for hepatocellular carcinoma. PMID- 14745323 TI - Evidence-based compression: prevention of stasis and deep vein thrombosis. AB - OBJECTIVE: To summarize the currently published scientific evidence for the venous flow effects of mechanical devices, particularly intermittent pneumatic compression, and the relation to prevention of deep vein thrombosis (DVT). SUMMARY BACKGROUND DATA: While intermittent pneumatic compression is an established method of DVT prophylaxis, the variety of systems that are available can use very different compression techniques and sequences. In order for appropriate choices to be made to provide the optimum protection for patients, the general performance of systems, and physiological effects of particular properties, must be analyzed objectively. METHODS: Medline was searched from 1970 to 2002, and all relevant papers were searched for further appropriate references. Papers were selected for inclusion when they addressed specifically the questions posed in this review. RESULTS: All the major types of intermittent compression systems are successful in emptying deep veins of the lower limb and preventing stasis in a variety of subject groups. Compression stockings appear to function more by preventing distension of veins. Rapid inflation, high pressures, and graded sequential intermittent compression systems will have particular augmentation profiles, but there is no evidence that such features improve the prophylactic ability of the system. CONCLUSIONS: The most important factors in selecting a mechanical prophylactic system, particularly during and after surgery, are patient compliance and the appropriateness of the site of compression. There is no evidence that the peak venous velocity produced by a system is a valid measure of medical performance. PMID- 14745324 TI - Split-liver transplantation in the United States: outcomes of a national survey. AB - OBJECTIVE: Assess application and outcomes of split-liver transplantation within the United States. SUMMARY BACKGROUND DATA: While a theoretically attractive mechanism to increase cadaver organ supply, split-liver transplantation has been infrequently applied. The American Society of Transplant Surgeons, in an attempt to gather preliminary data on split-liver transplantation, performed a data protected survey of transplant centers participating in the U.S. Scientific Registry for Transplant Recipients. METHODS: Between April 2000 and May 2001, 89 surgical teams were surveyed. Elicited data included graft type, recipient status, procurement method, graft sharing, graft outcomes, recipient outcomes, and experience with cadaver, whole-organ transplantation. RESULTS: Eighty-three surgical teams reported data on 207 left lateral segment, 152 right trisegment, 15 left lobe, and 13 right lobe grafts. The split procedure was performed ex vivo in 54% and in situ in 46% of grafts. Complications were frequent in all graft types with biliary and vascular complications equally distributed between grafts procured by either technique. Primary nonfunction, graft failure, and recipient death correlated with transplant status. CONCLUSIONS: Split-liver transplantation has been principally applied to adult-child pairs with at least one recipient critically ill. Biliary and vascular complications account for the majority of morbidity in grafts procured by either split technique with graft failure and recipient death observed more frequently in critically ill recipients. Enhanced utilization and improved results may be possible through improved information sharing and modification of allocation criteria. PMID- 14745325 TI - Safety zones for anterior abdominal wall entry during laparoscopy: a CT scan mapping of epigastric vessels. AB - OBJECTIVE: To determine the efficacy of CT scan in mapping the superior and inferior epigastric vessels, relative to landmarks apparent at laparoscopy. SUMMARY BACKGROUND DATA: Trauma to abdominal wall blood vessels occurs in 0.2% to 2% of laparoscopic procedures. Both superficial and deep abdominal wall vessels are at risk. The superficial vessels may be located by transillumination; however, the deep epigastric vessels cannot be effectively located by transillumination and, thus, other techniques should be used to minimize the risk of injury to these vessels. METHODS: Abdominal and pelvic CT images of 100 patients were studied. The location of the superior and inferior epigastric vessels from the midline were determined at five levels, correlated with each other and with the patient age, body mass index, and history of midline laparotomy using Pearson's correlation coefficient and multivariate analysis. RESULTS: CT scan was successful in mapping the epigastric vessels in 95% of patients. At the xiphoid process level, the superior epigastric vessels (SEA) were 4.41 +/- 0.13 cm from the midline on the right and 4.53 +/- 0.14 cm on the left. Midway between xiphoid and umbilicus, the SEA were 5.50 +/- 0.16 cm on the right of the midline and 5.36 +/- 0.16 cm on the left. At the umbilicus, the epigastric vessels were 5.88 +/- 0.14 cm on the right and 5.55 +/- 0.13 on the left of the midline. Midway between the umbilicus and symphysis pubis, the inferior epigastric (IEA) were 5.32 +/- 0.12 cm on right and 5.25 +/- 0.11 cm on the left. At the symphysis pubis, the IEA were 7.47 +/- 0.10 cm on the right and 7.49 +/- 0.09 cm away from the midline on the left side. CONCLUSIONS: Epigastric vessels are usually located in the area between 4 and 8 cm from the midline. Staying away from this area will determine the safe zone of entry of the anterior abdominal wall. PMID- 14745326 TI - Correlation between preoperative serum concentration of type IV collagen 7s domain and hepatic failure following resection of hepatocellular carcinoma. AB - OBJECTIVE: To determine the predictive value of the preoperative serum concentration of type IV collagen 7s domain (7s collagen) for postoperative hepatic failure in patients undergoing liver resection for hepatocellular carcinoma. SUMMARY BACKGROUND DATA: Clear and reliable criteria for predicting hepatic failure after liver resection are needed. The serum 7s collagen concentration correlates with the histologic degree of active hepatitis and hepatic fibrosis and may predict the regenerative potential of the liver. METHODS: Potential risk factors for postoperative hepatic failure, including the serum 7s collagen concentration, were evaluated in 251 patients who underwent liver resection for hepatocellular carcinoma. Prognostic significance was determined by univariate and multivariate analyses. RESULTS: Hepatic failure developed postoperatively in 25 patients, 4 of whom died. The serum 7s collagen concentration correlated with the histologic degree of hepatitis activity and hepatic fibrosis. The serum 7s collagen concentration was a risk factor for postoperative hepatic failure by univariate analysis and was the only risk factor on multivariate analysis. No patient with a serum 7s collagen concentration <12 ng/mL died of postoperative hepatic failure, and all 4 patients who died had a serum 7s collagen concentration >or=12 ng/mL. CONCLUSIONS: The preoperative serum 7s collagen concentration correlated independently with hepatic failure following liver resection for hepatocellular carcinoma. Patients whose serum 7s collagen is >or=12 ng/mL are poor candidates for hepatic resection. PMID- 14745327 TI - Abdominal drainage after hepatic resection is contraindicated in patients with chronic liver diseases. AB - OBJECTIVE: The aim of this study was to determine whether abdominal drainage is beneficial after elective hepatic resection in patients with underlying chronic liver diseases. SUMMARY BACKGROUND DATA: Traditionally, in patients with chronic liver diseases, an abdominal drainage catheter is routinely inserted after hepatic resection to drain ascitic fluid and to detect postoperative hemorrhage and bile leakage. However, the benefits of this surgical practice have not been evaluated prospectively. PATIENTS AND METHODS: Between January 1999 and March 2002, 104 patients who had underlying chronic liver diseases were prospectively randomized to have either closed suction abdominal drainage (drainage group, n = 52) or no drainage (nondrainage group, n = 52) after elective hepatic resection. The operative outcomes of the 2 groups of patients were compared. RESULTS: Fifty seven (55%) patients had major hepatic resection with resection of 3 Coiunaud's segments or more. Sixty-nine (66%) patients had liver cirrhosis and 35 (34%) had chronic hepatitis. Demographic, surgical, and pathologic details were similar between both groups. The primary indication for hepatic resection was hepatocellular carcinoma (n = 100, 96%). There was no difference in hospital mortality between the 2 groups of patients (drainage group, 6% vs. nondrainage group, 2%; P = 0.618). However, there was a significantly higher overall operative morbidity in the drainage group (73% vs. 38%, P < 0.001). This was related to a significantly higher incidence of wound complications in the drainage group compared with the nondrainage group (62% vs. 21%, P < 0.001). In addition, a trend toward a higher incidence of septic complications in the drainage group was observed (33% vs. 17%, P = 0.07). The mean (+/- standard error of mean) postoperative hospital stay of the drainage group was 19.0 +/- 2.2 days, which was significantly longer than that of the nondrainage group (12.5 +/- 1.1 days, P = 0.005). With a median follow-up of 15 months, none of the 51 patients with hepatocellular carcinoma in the drainage group developed metastasis at the drain sites. On multivariate analysis, abdominal drainage, underlying liver cirrhosis, major hepatic resection, and intraoperative blood loss of >1.5L were independent and significant factors associated with postoperative morbidity. CONCLUSION: Routine abdominal drainage after hepatic resection is contraindicated in patients with chronic liver diseases. PMID- 14745328 TI - The role of lymphadenectomy for liver tumors: further considerations on the appropriateness of treatment strategy. AB - OBJECTIVE: To evaluate the role of regional lymphadenectomy in patients with liver tumors. BACKGROUND: Lymph node status is 1 of the most important prognostic factors in oncologic surgery; however, the role of lymph node dissection remains unclear for hepatic tumors. METHODS: A total of 120 consecutive patients undergoing liver resections for primary and secondary hepatic tumors were prospectively enrolled in the study. "Regional" lymphadenectomy was carried out routinely after specimen removal. Incidence, site, and influence on survival of node metastases were analyzed. RESULTS: Only 1 postoperative complication (intra abdominal bleeding) was related to lymph node excision. Median number of dissected nodes was 6.8 +/- 3.6. Periportal, pericholedochal, and common hepatic artery stations were always removed. Lymph node metastases were found in 17 (16.5%) patients. The percentage rises to 20.3% when considering only noncirrhotic patients. The incidence of lymph node metastases was 7.5% for hepatocellular carcinoma, 14% for colorectal metastases, 40% for noncolorectal metastases, and 40% for intrahepatic cholangiocarcinoma (P < 0.002). Median survival time was 486 +/- 93.2 days among all patients with node metastases and 725 +/- 29.7 among patients without node metastases. The 2-year survival was 37.1% and 86.7%, in the 2 groups (P < 0.05). The 2-year recurrence rate was 77.6% and 45.3%, respectively (P < 0.05). CONCLUSIONS: Regional lymphadenectomy is a safe procedure after liver resection, and it should be routinely applied in patients with primary and secondary hepatic tumors, particularly in those without chronic disease. A careful evaluation of node status is nevertheless advisable also in patients with hepatocellular carcinoma on cirrhosis. PMID- 14745329 TI - Hepatic resection and transplantation for primary carcinoid tumors of the liver. AB - OBJECTIVE: To discuss the diagnosis and management of primary carcinoid tumors of the liver in light of our experience and a literature review. SUMMARY BACKGROUND DATA: Carcinoid tumors of the liver are rare and pose a diagnostic and management dilemma. This series is the largest reported and the only one to include liver transplantation as a treatment option. METHODS: Between March 1994 and May 2002, we treated 8 patients (4 male, 4 female) with primary hepatic carcinoid tumors. Carcinoid syndrome complicated only 1 of the cases. Treatment was by liver resection in 6 patients and orthotopic liver transplantation in 2. RESULTS: The diagnosis was confirmed histologically with light microscopy and immunohistochemistry in the absence of an alternative primary site. Six patients remain alive and disease free after follow-up of more than 3 years: 39, 43, 45, 50, 50, and 95 months. Two patients are recently postoperative. CONCLUSIONS: Active exclusion of an extrahepatic primary site is essential for the diagnosis of primary carcinoid of the liver. The mainstay of treatment should be liver resection, although liver transplantation may be considered in patients with widespread hepatic involvement. A radical surgical approach is warranted as this disease carries a better prognosis than for other primary hepatic tumors and for secondary hepatic carcinoids. PMID- 14745330 TI - Glutathione protects the rat liver against reperfusion injury after prolonged warm ischemia. AB - OBJECTIVE: To evaluate the potential of postischemic intravenous infusion of the endogenous antioxidant glutathione (GSH) to protect the liver from reperfusion injury following prolonged warm ischemia. BACKGROUND DATA: The release of reactive oxygen species (ROS) by activated Kupffer cells (KC) and leukocytes causes reperfusion injury of the liver after warm ischemia. Therefore, safe and cost-effective antioxidant strategies would appear a promising approach to prevent hepatic reperfusion injury during liver resection, but need to be developed. METHODS: Livers of male Lewis rats were subjected to 60, 90, or 120 minutes of normothermic ischemia. During a 120 minutes reperfusion period either GSH (50, 100 or 200 micromol/h/kg; n= 6-8) or saline (n= 8) was continuously administered via the jugular vein. RESULTS: Postischemic GSH treatment significantly prevented necrotic injury to hepatocytes as indicated by a 50-60% reduction of serum ALT and AST. After 1 hour of ischemia and 2 hours of reperfusion apoptotic hepatocytes were rare (0.50 +/- 0.10%; mean +/- SD) and not different in GSH-treated animals (0.65 +/- 0.20%). GSH (200 micromol GSH/h/kg) improved survival following 2 hours of ischemia (6 of 9 versus 3 of 9 rats; P < 0.05). Intravital fluorescence microscopy revealed a nearly complete restoration of sinusoidal blood flow. This was paralleled by a reduction of leukocyte adherence to sinusoids and postsinusoidal venules. Intravenous GSH administration resulted in a 10- to 40-fold increase of plasma GSH levels, whereas intracellular GSH contents were unaffected. Plasma concentrations of oxidized glutathione (GSSG) increased up to 5-fold in GSH-treated animals suggesting counteraction of the vascular oxidant stress produced by activated KC. CONCLUSIONS: Intravenous GSH administration during reperfusion of ischemic livers prevents reperfusion injury in rats. Because GSH is well tolerable also in man, this novel approach could be introduced to human liver surgery. PMID- 14745331 TI - Lymphatic drainage patterns from the breast. AB - OBJECTIVES: The aim of this study was to describe the lymphatic drainage patterns from the 5 "quadrants" of the breast. SUMMARY BACKGROUND DATA: Lymphatic mapping has provided techniques to visualize and harvest sentinel nodes in various locations and has generated renewed interest in nodes outside the axilla. METHODS: Between January 1997 and June 2002, 700 sentinel node procedures were performed in patients with cN0 breast cancer. Preoperative lymphoscintigraphy was performed after injection of 99mTc-nanocolloid into the tumor in a volume of 0.2 mL and a mean dose of 107.7 MBq (2.8 mCi). Intraoperatively, the sentinel node was pursued with the aid of a gamma-ray detection probe and patent blue dye (1.0 mL, into the lesion). The patients were divided into 5 groups according to the location of the primary breast cancer. In each group, a distinction was made between palpable and nonpalpable lesions of the breast. RESULTS: Drainage to either an axillary or an extra-axillary basin was observed in 678 patients. Both palpable and nonpalpable lesions may drain toward the internal mammary chain, although the latter more frequently, regardless of the quadrant. Drainage was also observed to supraclavicular, infraclavicular, interpectoral, and intramammary sentinel nodes. CONCLUSION: In each quadrant, a breast cancer may drain to sentinel nodes in various locations. There is a distinct difference in drainage patterns between palpable and nonpalpable lesions. These findings may improve the assessment of lymphatic dissemination in invasive breast cancer. PMID- 14745332 TI - The use of contrast-enhanced computed tomography before neoadjuvant chemotherapy to identify patients likely to be treated safely with breast-conserving surgery. AB - OBJECTIVE: To select suitable candidates for breast-conserving treatment (BCT) after neoadjuvant chemotherapy (NAC), based on the classification of tumors into localized or diffuse types using contrast-enhanced computed tomography (CE-CT). SUMMARY BACKGROUND DATA: A relatively high rate of loco-regional failure after BCT has been reported with breast cancer downstaged by NAC. Accurate assessment of the suitability of BCT and the response to NAC, before the initiation of NAC, will allow the optimal selection of an appropriate therapeutic course. METHODS: We evaluated 110 consecutive patients with operable breast carcinomas measuring 3 cm or more in diameter by CE-CT after NAC treatment with doxorubicin and docetaxel at National Cancer Center Hospital, Tokyo, from May 1998 to November 2001. Lesions were classified as either localized or diffuse types by mammography (MMG), ultrasonography (US), and CE-CT. RESULTS: Tumors designated as localized type by MMG, US, and CE-CT were reduced to tumors less than 3.0 cm (P < 0.0001) in a concentric circle (P < 0.0001). Localized tumors by CE-CT were treated safely with BCT maintaining a negative margin status (P = 0.01). In contrast, diffuse type tumors shrunk into a mosaic pattern consisting of tumors larger than 3.1 cm. Tumors classified as localized by CE-CT responded better pathologically than diffuse tumors (P = 0.0365). Multivariate analysis demonstrated that morphologic type by CE-CT and histologic type were significant predictors of candidates for safe BCT. CONCLUSIONS: The classification of tumors into either localized or diffuse types, using CE-CT before NAC administration, accurately predicts which tumors will be suitable candidates for BCT after NAC. PMID- 14745333 TI - Operative management of primary retroperitoneal sarcomas: a reappraisal of an institutional experience. AB - OBJECTIVE: To review our recent experience with primary retroperitoneal sarcomas, determine prognostic factors for disease recurrence and patient survival, and compare them to our previous results. BACKGROUND: Medical therapies have shown little efficacy in the management of retroperitoneal sarcomas, making total surgical extirpation the best chance for patient cure. METHODS: The case histories of all patients operated upon for retroperitoneal sarcomas between January 1983 and December 1995 were retrospectively reviewed. RESULTS: Ninety seven patients underwent attempted surgical resection of a primary retroperitoneal sarcoma. There were 54 (56%) men and 43 (44%) women, with a mean age of 59 years. Seventy-six (78%) patients underwent gross total resection, 13 (14%) had residual disease, and 8 (8%) underwent biopsy only with an actuarial 1 year survival of 88%, 51%, and 47%, respectively (P = 0.001). The actuarial 5- and 10-year survivals for patients who underwent gross total resection were 51% and 36%, respectively. Thirty-three patients (43%) developed locoregional recurrence, and 20 patients (26%) developed distant metastases at a median time of 12 months. The cumulative probability at 5 years was 44% for locoregional recurrence and 29% for distant metastases. On univariate analysis, factors associated with improved survival were complete resection of the tumor (P = 0.001), nonmetastatic disease at presentation (P = 0.01), low-grade tumors (P = 0.02), liposarcomas (P = 0.003), and no disease recurrence (P = 0.0001). Contrary to previous reports, the histologic subtype (P = 0.04) was the only significant factor predicting survival on multivariate analysis. CONCLUSIONS: Compared with our earlier experience, the rates of complete resection and overall survival have improved. Local control continues to be a significant problem in the management of retroperitoneal sarcomas. Because new surgical options for this problem are limited, further outcome improvement requires novel adjuvant therapies. PMID- 14745334 TI - Cecropin B enhances betalactams activities in experimental rat models of gram negative septic shock. AB - OBJECTIVE: To investigate the efficacy of imipenem, piperacillin combined with cecropin B in the prevention of lethality in 2 rat models of septic shock. Main outcome measures were bacterial growth in blood and intra-abdominal fluid, endotoxin and TNF-alpha concentrations in plasma, and lethality. BACKGROUND: Sepsis remains a serious clinical problem despite intense efforts to improve survival. It is a major cause of morbidity and mortality in hospitalized patients. The primary cause of Gram-negative shock results from activation of host effector cells by endotoxin, the lipopolysaccharide (LPS) associated with cell membranes of Gram-negative bacteria. METHODS: Adult male Wistar rats were given (1). an intraperitoneal injection of 1 mg of Escherichia coli 0111:B4 LPS or (2). 2 x 1010 CFU of E. coli ATCC 25922. All animals were randomized to receive intraperitoneally isotonic sodium chloride solution, 1 mg/kg cecropin B, 20 mg/kg imipenem, and 120 mg/kg piperacillin alone and combined with 1 mg/kg cecropin B. Each group included 20 animals. RESULTS All compounds reduced the lethality when compared with controls. Piperacillin and imipenem significantly reduced the lethality and the number of E. coli in abdominal fluid compared with saline treatment. On the other hand, each betalactam determined an increase of plasma endotoxin and TNF-alpha concentration. Combination between cecropin B and betalactams showed to be the most effective treatment in reducing all variables measured. CONCLUSION: Cecropin B enhances betalactams activities in Gram-negative sepic shock rat models. PMID- 14745335 TI - Enteral administration of high-fat nutrition before and directly after hemorrhagic shock reduces endotoxemia and bacterial translocation. AB - OBJECTIVE: To determine whether potential enhancement of endotoxin neutralization via high-fat enteral nutrition affects endotoxemia and bacterial translocation after hemorrhage. SUMMARY BACKGROUND DATA: Endotoxin and bacterial translocation due to gut barrier failure are important initiating events in the pathogenesis of sepsis after hemorrhage. Systemic inhibition of endotoxin activity attenuates bacterial translocation and distant organ damage. Triacylglycerol-rich lipoproteins constitute a physiological means of binding and neutralizing endotoxin effectively. We hypothesized that enhancement of triacylglycerol-rich lipoproteins via high-fat enteral nutrition would reduce endotoxemia and prevent bacterial translocation. METHODS: A rat model of nonlethal hemorrhagic shock was used. Hemorrhagic shock (HS) rats were divided into 3 groups: rats starved overnight (HS-S); rats fed with a low-fat enteral diet (HS-LF), and rats receiving a high-fat enteral diet (HS-HF). RESULTS: Circulating triacylglycerol and apolipoprotein B, reflecting the amount of triacylglycerol-rich lipoproteins, were elevated in HS-HF rats compared with both HS-S rats (P 2 cm), grade III and IV esophagitis, and Barrett's esophagus. The following were assessed over a follow-up period of 6 months: GERD-Health-Related Quality of Life, Gastrointestinal Symptom Rating Scale, and SF-36 Health Survey, medication use, 24-hour esophageal pH monitoring and esophageal manometry. Patients underwent a single, full-thickness plication in the gastric cardia just distal to the gastroesophageal junction. Re-treatment was not permitted. RESULTS: A total of 64 patients (mean age 46.3 years, range 23-71 years) underwent endoscopic full thickness plication (mean procedure time 17.2 minutes). At 6 months after plication, proton pump inhibitor therapy had been eliminated in 74% of previously medication-dependent patients. Median GERD-Health-Related Quality of Life scores improved 67% (19.0 vs. 5.0; p<0.001). Improvements also were observed in median Gastrointestinal Symptom Rating Scale and SF-36 Health Survey mental and physical composite scores. Median esophageal acid exposure improved significantly (10 vs. 8; p<0.008) with normalization of pH noted in 30% of patients. No significant change in esophageal manometry was noted. One gastric perforation occurred and was managed conservatively without sequelae. CONCLUSIONS: In this study, a single full-thickness plication placed at the gastroesophageal junction reduced symptoms, medication use, and esophageal acid exposure associated with GERD. PMID- 14745388 TI - Nasogastric aspirate predicts high-risk endoscopic lesions in patients with acute upper-GI bleeding. AB - BACKGROUND: Active upper-GI bleeding (spurting or oozing) or a visible vessel at endoscopy are high-risk lesions that predict recurrence of bleeding. The aim of this study is to determine whether nasogastric aspirate predicts the presence of high-risk lesions. METHODS: The Canadian Registry of patients with Upper Gastrointestinal Bleeding undergoing Endoscopy was used to identify patients with upper-GI bleeding who underwent nasogastric aspiration and subsequent endoscopy. An association between nasogastric aspirate findings (bloody, "coffee ground," clear/bile) and high-risk lesions was sought. RESULTS: Of 1869 patients in the registry, 520 had documented nasogastric aspiration before endoscopy. Those who underwent aspiration did not differ from those who did not. A bloody nasogastric aspirate was significantly associated with high-risk lesions (odds ratio 4.82: 95% CI[2.3, 10.1] vs. clear/bile; and odds ratio 2.8: 95% CI[1.8, 4.3] vs. coffee ground). A bloody nasogastric aspirate had the highest specificity for high-risk lesions (75.8%: 95% CI[70.0, 80.0]) with a negative predictive value of 77.9%: 95% CI[73.2, 82.0], and raised the probability of having a high-risk lesions from 0.29 to 0.45. A clear nasogastric aspirate reduced the likelihood to 0.15. Nasogastric aspirate yielded the most useful information in hemodynamically stable patients without hematemesis. CONCLUSIONS: Nasogastric aspirate is useful in predicting high-risk lesions. Whether it can be used to determine which patients would benefit from earlier endoscopy deserves further study. PMID- 14745389 TI - The effect of lidocaine sprayed on the major duodenal papilla on the frequency of post-ERCP pancreatitis. AB - BACKGROUND: Acute pancreatitis remains a serious cause of ERCP-related morbidity. Topical application of lidocaine reportedly blunts cholecystokinin release from intestinal mucosa and reduces sphincter of Oddi spasm. A randomized trial was conducted to evaluate the effect of lidocaine sprayed on the major duodenal papilla on the frequency of post-ERCP pancreatitis. Secondary outcomes evaluated were ease of cannulation and severity of post-ERCP pancreatitis. METHODS: Patients undergoing ERCP were randomized in blocks of 6 to have 10 mL of either 1% lidocaine or normal saline solution sprayed on the major papilla before cannulation. Patients were observed for the development of post-ERCP pancreatitis. Patient history- and procedure-related variables were recorded. RESULTS: A total of 326 patients were enrolled, of whom 32 were excluded after randomization but before analysis. Of patients analyzed, 145 were randomized to treatment with lidocaine and 149 to placebo. No patient was lost to follow-up. There was no significant difference noted in patient history- or procedure related variables. Seven patients in the lidocaine group and 5 in the placebo group developed post-ERCP pancreatitis (p=0.73). Ease of cannulation did not differ between the two groups. CONCLUSIONS: Lidocaine sprayed on the major papilla does not decrease the frequency of post-ERCP pancreatitis. PMID- 14745390 TI - EUS-guided trucut needle biopsies in patients with solid pancreatic masses: a prospective study. AB - BACKGROUND: A trucut needle biopsy device that can be used to obtain specimens from the pancreas and other perigastric organs under EUS guidance has been developed and successfully tested in animals. Moreover, EUS-guided trucut needle biopsy has been used safely in humans and appears to provide more accurate results than EUS-guided FNA. This study prospectively assessed the clinical utility of this new device in patients with solid pancreatic masses. METHODS: Twenty-three consecutive patients with radiologically detected solid pancreatic masses underwent EUS-guided trucut needle biopsy. Pancreatic malignancy detected by EUS-guided trucut needle biopsy was considered a definitive diagnosis. Further diagnostic procedures and clinical course were used to establish or exclude the presence of malignancy in all other patients. RESULTS: Pancreatic tissue was obtained in 17 of the 23 patients (74%), including all patients in whom the transgastric approach was used. No acute or long-term complication was observed. Histopathologic evaluation revealed pancreatic cancer in 12 patients. CT-guided biopsy specimens were obtained in 4 of the 5 patients with a negative EUS-guided trucut needle biopsy result; two were positive for adenocarcinoma. Overall diagnostic accuracy was 61%. Subgroup analysis of the 16 patients in whom EUS guided trucut needle biopsy was successful and who were available for follow-up revealed a diagnostic accuracy of 87.5%. CONCLUSIONS: This prospective study demonstrates that EUS-guided trucut needle biopsy, when performed transgastrically, is safe and accurate in the evaluation of patients with solid pancreatic masses. PMID- 14745391 TI - Evaluation of in vivo endoscopic autofluorescence spectroscopy in gastric cancer. AB - BACKGROUND: The aim of this study was to evaluate light-induced autofluorescence spectroscopy for the in vivo diagnosis of gastric cancer. METHODS: A total of 344 endogenous fluorescence spectra were obtained from normal (164) and cancerous gastric mucosa (180) in 15 patients with pure adenocarcinoma and in 16 patients with gastric cancer containing signet-ring cells. A special light source capable of delivering either white or violet-blue light for the excitation of tissue autofluorescence via the endoscope was used. Endogenous fluorescence spectra emitted by the tissue were collected with a fiberoptic probe and analyzed with a spectrograph. RESULTS: Gastric adenocarcinoma exhibits specific changes in the emitted fluorescence spectra as compared with normal gastric mucosa. By algorithmic classification of the spectra, a sensitivity of 84%, specificity of 87%, a likelihood ratio for a positive test of 6.5 and for a negative test of 0.18 were obtained for the diagnosis of pure adenocarcinoma of the stomach. However, gastric cancer with signet-ring cells exhibits great variation in emitted autofluorescence spectra as compared with normal mucosa. The sensitivity for the diagnosis of all carcinomas containing signet-ring cells was 55%, specificity 85%, the likelihood ratio for a positive test was 3.7 and for a negative test, 0.53. The diagnostic value decreases with increasing numbers of signet-ring cells and tumor grade. CONCLUSIONS: Light-induced autofluorescence spectroscopy is a new and promising bio-optical technique for the endoscopic in vivo diagnosis of gastric adenocarcinoma. The poor diagnostic accuracy for signet ring cell carcinoma may be explained by the diffuse and frequent submucosal growth of this tumor and the presence of collagen fibers. PMID- 14745392 TI - EMR combined with chemoradiotherapy: a novel treatment for superficial esophageal squamous-cell carcinoma. AB - BACKGROUND: Esophagectomy or chemoradiotherapy (CRT) are the procedures of choice for patients with superficial esophageal squamous-cell carcinoma. However, esophagectomy is highly invasive, and CRT is associated with the risk of local failure. A study was conducted of a novel treatment, EMR combined with CRT, for patients with superficial esophageal carcinoma. EMR was performed for the purpose of complete local tumor control and chemoradiotherapy was performed for regional and distant control. METHODS: EMR combined with CRT was performed for patients with esophageal carcinoma invading the muscularis mucosae or upper submucosa who refused esophagectomy. The planned treatment after EMR was 40 to 46 Gy of external beam radiation to the mediastinum, including the supraclavicular fossa or cardia. Chemotherapy was given during weeks 1 and 5 (5-fluorouracil, 700 mg/m(2) per 24 hours in a 120-hour infusion, and cisplatin 15 mg/m(2) per day intravenously on days 1 to 5). RESULTS: During the study period, 16 patients underwent EMR combined with CRT (EMR plus CRT group) and 39 patients with similar stage cancer underwent esophagectomy (surgical resection group). None of the patients in the EMR plus CRT group have had local recurrence or metastasis. Overall survival rates at 5 years in the EMR plus CRT and surgical resection groups were estimated to be, respectively, 100% and 87.5%. CONCLUSIONS: Although this study was not randomized, the results suggest that EMR combined with CRT is a safe and effective method for treating patients with superficial esophageal carcinoma. The results were equivalent or, in view of the lower degree of invasiveness, superior to surgical resection. PMID- 14745393 TI - Accuracy of EUS criteria and primary tumor site for identification of mediastinal lymph node metastasis from non-small-cell lung cancer. AB - BACKGROUND: EUS with FNA is useful for staging non-small-cell lung cancer. However, benign mediastinal adenopathy is common. The aims of this study were to identify clinical factors, especially primary tumor location, and EUS lymph nodal characteristics predictive of aortopulmonary window and subcarinal lymph node metastases of non-small-cell lung cancer. METHODS: Patients with known or suspected non-small-cell lung cancer underwent EUS staging at which EUS-FNA was performed for all identified mediastinal lymph nodes. Clinical characteristics, primary tumor data, EUS findings, and histopathology were reviewed. Exact tests were performed for both aortopulmonary window and subcarinal lymph nodes to identify factors predictive of malignant cytology. RESULTS: Ninety-two patients with non-small-cell lung cancer were included. Fifty-one had aortopulmonary window, and 73 had subcarinal lymph nodes on EUS. The EUS with FNA specimens were interpreted as suspicious or diagnostic for malignancy for 9 aortopulmonary window and 9 subcarinal lymph nodes. When comparing benign vs. malignant EUS with FNA findings for aortopulmonary window and subcarinal lymph nodes, only lymph node size of 1 cm or greater and sharp lymph nodal edges were associated with malignancy in lymph nodes at both sites, whereas primary tumor site, lymph node shape, and echogenicity were associated with malignant subcarinal nodes. When 4 classic lymph nodal features of malignancy were evaluated, the presence of 3 or more typical features had positive and negative predictive values of, respectively, 41% and 96%. CONCLUSIONS: Although tumor location and EUS lymph nodal characteristics are associated with malignant involvement of lymph nodes, the accuracy of these predictors does not obviate the need for cytologic evaluation. EUS with FNA should be performed for all lymph nodes when an abnormal finding will alter management. PMID- 14745394 TI - Clinicopathologic and endoscopic features of colorectal serrated adenoma: differences between polypoid and superficial types. AB - BACKGROUND: Serrated adenoma is a distinct histologic colorectal lesion. There are two macroscopic types: polypoid and superficial. The aim of this study was to clarify clinicopathologic and endoscopic differences between polypoid and superficial serrated adenomas. METHODS: An analysis was conducted of the clinicopathologic and endoscopic features for 240 polypoid and 127 superficial serrated adenomas examined by colonoscopy, and the surface pit patterns of 114 polypoid and 64 superficial serrated adenomas examined by magnifying videoendoscopy. RESULTS: The male:female gender ratio for the polypoid serrated adenomas (3.5:1) was significantly higher than that for the superficial serrated adenomas (1.7:1). Superficial serrated adenomas were significantly larger than polypoid serrated adenomas (mean [standard deviation], respectively, 10.1 [7.9] mm vs. 6.3 [4.6] mm). In the distal segments of the colorectum, polypoid serrated adenomas were more common than superficial serrated adenomas. Granulonodular and lobular appearances at endoscopy were significantly more common for polypoid (23.3%) than for superficial serrated adenomas (7.1%). Pit patterns differed between the lesion types: polypoid serrated adenomas had type III(L) or IV pit patterns; all superficial serrated adenomas had the type II pit pattern. The relative frequency of occurrence of high-grade dysplasia and carcinoma in situ among superficial serrated adenomas (25.2%) was significantly greater than that among polypoid serrated adenomas (9.2%). The tubulovillous growth pattern was significantly more common in polypoid tumors (31.5%) than in superficial tumors (0%). CONCLUSIONS: Polypoid and superficial serrated adenomas have different clinicopathologic characteristics and growth patterns. PMID- 14745395 TI - A new method of EMR: submucosal injection of a fibrinogen mixture. AB - BACKGROUND: The technical limitation associated with submucosal injection of normal saline solution during EMR is the relatively short duration of mucosal elevation. The clinical application of a new method of EMR with submucosal injection of a fibrinogen mixture was evaluated. METHODS: Thirty-five early stage neoplastic gastric lesions were resected by EMR with submucosal injection of a fibrinogen mixture. The efficacy and clinical outcomes were analyzed. RESULTS: Additional submucosal injection was not required for any of the 35 lesions to complete the EMR with submucosal injection of a fibrinogen mixture procedure. The rates of en bloc resection and complete resection were, respectively, 82.9% and 88.6%. The en bloc resection rate was significantly lower for lesions over 20 mm in diameter (60% vs. 92%; p<0.05) and for lesions on the lesser curvature or posterior wall of the stomach compared with those on the greater curvature or anterior wall (55.6% vs. 92.3%; p<0.05). The rate of complete resection also was dependent on the size and location of the lesions. There was no major EMR with submucosal injection of a fibrinogen mixture related complication including bleeding or perforation. CONCLUSIONS: EMR with submucosal injection of a fibrinogen mixture is an easy, safe, and technically efficient method for complete EMR. PMID- 14745396 TI - Endoscopic management of adenoma of the major duodenal papilla. AB - BACKGROUND: It is well established that adenoma of the major duodenal papilla has a potential for malignant transformation. Standard treatment has been surgical (duodenotomy/local resection, pancreaticoduodenectomy). Endoscopic management is described, but there is no established consensus regarding the approach to papillectomy or the need for surveillance. This study describes endoscopic management and long-term follow-up of papillary tumors by 4 groups of expert pancreaticobiliary endoscopists. METHODS: Consecutive patients with papillary tumors referred to 4 pancreaticobiliary endoscopy centers for evaluation for endoscopic papillectomy were reviewed. For each patient, an extensive questionnaire was completed, which included 19 preoperative and 15 postoperative data points. A total of 103 patients (53 women, 50 men, age range 24-93) who underwent attempted endoscopic resection were included. Of these, 72 had sporadic adenoma, and the remaining patients had familial adenomatous polyposis, including Gardner's variant. Presenting symptoms were jaundice/cholangitis/pain (n=59), pancreatitis (n=18), and bleeding (n=12). Twenty-six patients were asymptomatic. RESULTS: Endoscopic treatment was successful, long term, in 83 patients (80%) and failed (initial failure or recurrent tumor) in 20 (20%) patients. Success was significantly associated with older age (54.7 [16.6] vs. 46.6 [21.7] years; p=0.08) and smaller lesions (21.1 [8.3] vs. 29.7 [7.2] mm; p<0.0001). Success rate was higher for sporadic lesions compared with genetically determined lesions (63 of 72 [86%] vs. 20 of 31 [67%]; p=0.02). There were 10 initial failures, which was more common for sporadic lesions (7 of 10). The overall success rate for papillectomy was similar in patients who had adjuvant thermal ablation (81%) compared with those who did not (78%). However, recurrence (n=10) was more common in the former group (9 of 10, [90%]; p=0.22). Complications (n=10) included acute pancreatitis (n=5), bleeding (n=2), and late papillary stenosis (n=3). Acute pancreatitis was more common in patients who did not have pancreatic duct stents placed (17% vs. 3.3%). Papillary stenosis was more frequent without short-term pancreatic duct stent placement (15.4% vs. 1.1%), although the difference was not statistically significant, because this complication was infrequent. CONCLUSIONS: Endoscopic treatment of papillary adenoma in selected patients appears to be highly successful. The majority can undergo complete resection after ERCP. In expert hands, complications are infrequent and may be avoided by routine placement of a pancreatic duct stent. PMID- 14745397 TI - Endoscopic therapy for upper-GI vascular ectasias. AB - BACKGROUND: Upper-GI vascular ectasias, including angiodysplasia and gastric antral vascular ectasia may present with either acute or chronic bleeding. Endoscopic thermal modalities have been used to control acute bleeding and reduce transfusion requirements. METHODS: Endoscopic experience was reviewed for a 6 year period during which 32 patients requiring blood transfusions for upper-GI angiodysplasia or gastric antral vascular ectasia were evaluated. Patients seen during the first 5 years were treated with either Nd:YAG laser photocoagulation or multipolar electrocoagulation. During the most recent 12 months, all patients were treated by argon plasma coagulation. Response to therapy was assessed by change in mean Hb and transfusion requirements. RESULTS: Overall, 16 patients were treated by laser photoablation alone; 9, argon plasma coagulation with or without laser; and 7, multipolar electrocoagulation with or without laser. Mean follow-up for all patients was 19 months. After therapy, mean Hb concentration rose from 76 to 114 g/L for patients with gastric antral vascular ectasia and from 85 to 118 g/L for those with angiodysplasia. Endoscopic therapy abolished or reduced transfusion requirements in 93% of patients with gastric antral vascular ectasia and 76% with angiodysplasia. Patients with gastric antral vascular ectasia required a mean of 6 treatment sessions, while those with angiodysplasia required one to two sessions. CONCLUSIONS: Endoscopic thermal ablation effectively controls acute bleeding and reduces transfusion requirements in most patients with upper-GI vascular ectasias. Patients with gastric antral vascular ectasia require significantly more treatment sessions to achieve this effect. PMID- 14745398 TI - Clinical and sociodemographic factors associated with colon surveillance among patients with a history of colorectal cancer. AB - BACKGROUND: Substantial variability in the use of colon surveillance among colorectal cancer survivors has been reported. This study sought to examine trends in the use of colon surveillance among patients who have had colorectal cancer and to investigate factors associated with utilization. METHODS: Health maintenance organization enrollees with a diagnosis of local or regional colon or rectal cancer between January 1993 and December 1999 were studied. Receipt of a colon examination by colonoscopy or by flexible sigmoidoscopy, together with barium contrast radiography of the colon was determined from automated clinical records, and rates of colon surveillance were estimated by using survival analysis. RESULTS: A total of 1002 patients with a diagnosis of colorectal cancer met inclusion criteria for the study. Colon examinations were performed in 61% of patients within 18 months of diagnosis and in 80% of patients within 5 years of diagnosis. The median time from diagnosis to first colon surveillance examination (14 months) was unchanged over the study period, but the interval between first and second surveillance examinations increased by 17 months (p<0.001). Patients over 80 years of age (relative risk=0.32; 95% CI[0.22, 0.45]) and those with rectal cancer (relative risk=0.80; 95% CI[0.66, 0.97]) were less likely to undergo surveillance. Higher socioeconomic status (relative risk=1.29; 95% CI[1.03, 1.61]) and being married (relative risk=1.27; 95% CI[1.05, 1.53]) were associated with greater utilization. There was lower utilization among African American patients (relative risk=0.70; p=0.14) and increased utilization among other minorities (relative risk=1.47; p=0.06). CONCLUSIONS: There is substantial variability in the use of colon examination for surveillance in patients with a history of colorectal cancer, and clinical and sociodemographic factors appear to influence the likelihood of surveillance. PMID- 14745399 TI - Ureterosigmoid anastomosis: risk of colorectal cancer and implications for colonoscopists. PMID- 14745400 TI - The venerable nasogastric tube. PMID- 14745401 TI - Colonic lymphomatous polyposis. PMID- 14745402 TI - Bleeding jejunal diverticulum. PMID- 14745403 TI - Chicken-bone ulcer. PMID- 14745404 TI - Colocolic fistula. PMID- 14745405 TI - Recurrent esophageal tears. PMID- 14745406 TI - Ileal Dieulafoy's lesion. PMID- 14745407 TI - Endoclips for GI endoscopy. PMID- 14745408 TI - EUS-guided portal vein catheterization and pressure measurement in an animal model: a pilot study of feasibility. AB - BACKGROUND: The extrahepatic portal vein is inaccessible to direct catheterization. METHODS: Because EUS can readily image the portal vein, the feasibility of EUS-guided portal vein catheterization by using a 22-gauge needle was studied in 7 normal pigs and 14 pigs in which portal hypertension was induced (7/14 anticoagulated). RESULTS: Catheterization was not possible by EUS or transhepatic methods in, respectively, 3 and 5 animals. One anticoagulated animal had a small amount of periduodenal bleeding as a result of EUS catheterization. The mean normal portal vein pressure (1 standard deviation) as determined by EUS and transhepatic methods was, respectively, 20.3 (4) mm Hg and 20.4 (2) mm Hg. Injection of polyvinyl alcohol particles increased the portal vein pressure by 10.2 (11.59) mm Hg. There was a close correlation under all conditions between the mean portal vein pressures obtained by EUS and transhepatic catheterization (r=0.91). CONCLUSIONS: EUS-guided portal vein catheterization appears to be feasible in an animal model and provides accurate pressure measurements. PMID- 14745409 TI - Percutaneous traction-assisted EMR by using an insulation-tipped electrosurgical knife for early stage gastric cancer. AB - BACKGROUND: EMR now is accepted as a minimally invasive treatment for early stage gastric cancer. However, the endoscopic approach sometimes is limited by the size and location of the tumor. The technique and early results of percutaneous, traction-assisted EMR for resection of larger early stage gastric cancers is described. METHODS: EMR, by using an insulation-tipped electrosurgical knife, was performed with the simultaneous assistance of laparoscopy devices. A small snare was introduced into the gastric lumen through a gastric port (2-mm diameter) to grasp and pull the EGC away from the muscularis propria to facilitate resection. RESULTS: The technique was performed successfully in our endoscopic suite in the first patient under general anesthesia and the remaining seven under conscious sedation. Tumors situated in any part of the stomach could be reached with percutaneous assistance, and this facilitated en bloc resection of large specimens (mean size 50 mm). The procedure was performed in about 60 minutes. Seven patients (88%) resumed eating 1 or 2 days after surgery. All patients were discharged after 1 week without complication. CONCLUSIONS: Percutaneous, traction assisted EMR is useful for resection of large lesions en bloc and can be performed easily and safely under direct vision. PMID- 14745410 TI - Narrow-band imaging system with magnifying endoscopy for superficial esophageal lesions. AB - BACKGROUND: By assessing the intrapapillary capillary loop in esophageal mucosa, magnifying endoscopy can play an important role in the evaluation of superficial esophageal lesions. A newly developed narrow-band imaging system was applied to magnifying endoscopy in a clinical setting; the benefit of the narrow-band imaging system was evaluated. METHODS: Forty-one patients (37 men, 4 women; mean age 63.5 [7.3] years) were enrolled between March 2002 and January 2003 in the study. Endoscopy was performed with a magnifying endoscope, a standard video endoscopic system, and a narrow-band imaging system. The assessment consisted of 3 phases: a numerical analysis of the red, green, blue color value of endoscopic images, creation of model images, and assessment on the actual images. In the numerical analysis, the red, green, blue color value for intrapapillary capillary loop and background mucosa were obtained, and the ratio and contrast value were calculated. RESULTS: In the numerical analysis, both the ratio and the contrast value between the intrapapillary capillary loop and background mucosa were statistically different. Based on an evaluation of created model images, almost all assessors found the narrow-band imaging system to be superior. In the assessment of actual images, the narrow-band imaging system improved overall accuracy for depth of invasion, especially for inexperienced endoscopists. CONCLUSIONS: The narrow-band imaging system improved the accuracy of magnifying endoscopy for assessment of esophageal lesion. PMID- 14745411 TI - Endoscopic management of gastrocutaneous fistula after bariatric surgery by using a fibrin sealant. AB - BACKGROUND: Gastrocutaneous fistula is an uncommon and difficult to treat complication that occurs in 0.5% to 3.9% of patients who undergo gastric surgery. Sepsis usually follows, and, when it is not managed effectively, the associated mortality rate can be as high as 85%. A fibrin sealant was used to endoscopically manage gastrocutaneous fistulas that developed in 3 morbidly obese patients after bariatric surgery. METHODS: Two of 14 (14.29%) patients who underwent vertical gastroplasty (MacLean procedure) developed a non-healing gastrocutaneous fistula. In addition, one of 24 (4.17%) patients who had a biliopancreatic diversion with preservation of pylorus developed a gastrocutaneous fistula. Endoscopic application of a fibrin sealant was performed under direct vision via a double lumen catheter passed through a forward-viewing endoscope. OBSERVATIONS: Treatment was successful in all patients after one or more endoscopic sessions in which the fibrin sealant was applied; no evidence of fistula was found at follow up endoscopy. CONCLUSIONS: Endoscopic closure of gastrocutaneous fistula with human fibrin tissue sealant is simple, safe, and effective, and, in some cases, can be life-saving. Endoscopic application of fibrin sealant should be considered a therapeutic option for treatment of gastrocutaneous fistula that develops after bariatric surgery. PMID- 14745412 TI - Unusual presentations of aorto-enteric fistula. AB - BACKGROUND: Aorto-enteric fistula is rare but can result in exsanguination without timely surgery or endovascular stent placement. METHODS: Four cases of aorto-enteric fistula were reviewed in which the presentation was unusual and diagnosis difficult. OBSERVATIONS: The first patient had an aorto-sigmoid fistula in the setting of an aorto-bi-femoral graft. Two patients had a primary aorto enteric fistula, one to the stomach from a suprarenal aortic aneurysm, and the other, to the duodenum in the setting of retroperitoneal spread of renal cancer. The aortoduodenal fistula recurred in the 4th patient within 3 months of surgical repair; this patient is the only one who survived long term. CONCLUSIONS: When presentation is atypical, the diagnosis of aorto-enteric fistula can be extremely difficult. Because investigative studies are not consistently useful in making a definitive pre-operative diagnosis, a strong index of clinical suspicion and a willingness to consider surgical exploration are essential for timely and successful management. PMID- 14745413 TI - Extramedullary plasmacytoma of the rectum arising in ulcerative colitis: case report and review. PMID- 14745414 TI - EUS-guided FNA of a thyroid mass. PMID- 14745415 TI - Endoscopic therapy for bleeding proximal esophageal varices: a case report. PMID- 14745416 TI - The value of capsule endoscopy in the diagnosis and management of Crohn's disease: report of two cases. PMID- 14745417 TI - Metastatic gastric carcinoma presenting as multiple submucosal colonic cysts. PMID- 14745418 TI - Submucosal nodules containing purulent fluid: a colonoscopic sign of actinomycosis? PMID- 14745419 TI - Endoscopic removal of a metallic biliary stent: case report. PMID- 14745420 TI - Use of a duodenoscope to identify and treat a colonic vascular malformation. PMID- 14745421 TI - Laparoscopy-assisted transgastrostomy ERCP after bariatric surgery: case report of a novel approach. PMID- 14745424 TI - Myelodysplastic syndromes. AB - Myelodysplastic syndrome (MDS) is a clonal disorder characterized by ineffective hematopoiesis, which can lead to either fatal cytopenias or acute myelogenous leukemias (AML). During the last 15 years, important progress has been made in the understanding of the biology and prognosis of MDS. Risk-adapted treatment strategies were established due to the high median age (60-75 years) of MDS patients and the individual history of the disease (number of cytopenias, cytogenetic changes, transfusion requirements). The use of allogeneic bone marrow transplantation for MDS patients currently offers the only potentially curative treatment, but this form of therapy is not available for the 'typical' MDS patient who is >60 years of age. The development of small molecules directed against specific molecular targets with minimal adverse effects is the hope for the future. Innovative uses of immunomodulatory agents and the optimizing of cytotoxic treatment should continue to help in the treatment of MDS. PMID- 14745425 TI - Strategies to improve the outcome of stem cell transplantation in multiple myeloma. AB - Multiple myeloma (MM) is an incurable hematological malignancy with an average survival of 3 years with conventional therapy. Allogeneic hematopoietic cell transplantation (allo-HCT) may cure some patients, but has been associated with a very high transplantation-related mortality (TRM) of over 40%.(1) In contrast to allo-HCT, autologous hematopoietic cell transplantation (AHCT) has been much safer, with a TRM <3% in the 1990s. Therefore, in the last 15 years AHCT has become a common procedure for MM patients. The widespread use of AHCT has been associated with a median survival of 55-72 months,(2,3,4,5,6) and two large randomized trials have shown that AHCT is superior to conventional chemotherapy for the treatment of MM.(3,7) Approaches to improve the outcome of stem cell transplantation for MM patients include consideration of patient status, efficacy and toxicity of induction therapy, source of hematopoietic rescue, conditioning regimens, and maintenance therapy. Recent attempts to improve outcome include tandem AHCT, AHCT followed by RIC (reduced intensity conditioning) allo-HCT, and allo-HCT with T-cell depletion and subsequent donor-lymphocyte infusions (DLI), while novel therapies and improved supportive care may improve the overall survival (OS) of all MM patients with or without transplantation. PMID- 14745426 TI - Possibility of long-term remission in patients with advanced hematologic malignancies after reduced intensity conditioning regimen (RIC) and allogeneic stem cell transplantation. AB - High-dose chemotherapy and radiation used as a preparative regimen for allogeneic stem cell transplantation (SCT) produces a considerable morbidity and mortality. An alternative strategy, developed to reduce transplant-related toxicity and to induce graft versus malignancy effect in the presence of full hematopoietic engraftment, includes the application of RIC that provide sufficient immunosuppression. We studied the efficacy and toxicity of RIC followed by allogeneic SCT in elderly patients or with relative contraindications to conventional SCT. In all, 22 patients with hematologic malignancies and HLA identical sibling donors were included in this study. All patients were either refractory to therapy or beyond first complete remission (CR). The majority of patients received fludarabine 120 mg/m(2)+thiotepa 10 mg/kg as conditioning regimen and cyclosporine as graft versus host disease (GVHD) prophylaxis. Organ toxicity was acceptable and all evaluable patients achieved engraftment. Three patients (15%) showed grade >II aGVHD; extensive chronic GVHD occurred in three out of 15 evaluable patients (20%). With median follow-up of 63.5 months, survival was 31%, disease-free survival (DFS) was 23%. All the durable responses occurred in patients who developed GVHD. Transplant related mortality (TRM) at 100 days was 27.3%, 67% of that caused by infections, while 6-year cumulative incidence of TRM was 38%. Our data show that RIC: (1). allows the engraftment of HLA-matched hematopoietic stem cells (2). provide durable responses in patients not eligible for conventional SCT exploiting the graft-versus-malignancy effect and (3). is loaded by an high rate of fatal infections. PMID- 14745427 TI - Non-transferrin-bound serum iron (NTBI) in megaloblastic anemia: effect of vitamin B(12) treatment. AB - INTRODUCTION: The abnormalities in iron metabolism associated with megaloblastic anemia are rapidly reversed by B(12) therapy in pernicious anemia (PA). Although non-tranferrin-bound plasma iron (NTBI) was previously shown to be associated with severe iron overload, its origin is unknown. METHODS AND RESULTS: Four patients with PA were studied before and after B(12) treatment. NTBI was measured by a fluorescence-based one-step assay. All patients had very high transferrin saturation, NTBI values ranging from 1.1 to 2.6 micromol/l and normal serum ferritins. B(12) treatment resulted in the disappearance of NTBI and normalization of transferrin saturation within 22-42 h. CONCLUSIONS: The prompt disappearance of NTBI following B(12) therapy implicates catabolic iron derived from ineffective erythropoiesis as the major source of NTBI in untreated PA and possibly in thalassemia major and sideroblastic anemia. Our findings offer further insight into the pathogenesis of NTBI in diseases associated with abnormal erythropoiesis. PMID- 14745428 TI - Comparative study of two portable systems for oral anticoagulant monitoring. AB - Portable prothrombin time (PT) monitors offer the potential for both simplifying and improving oral anticoagulation management. It is necessary to evaluate their concordance and correlation with other PT systems. Our objective was to evaluate the concordance and clinical correlation of two portable PT determination systems, ProTime (ITC) and CoaguChek S (Roche Diagnostics). In all, 20 healthy individuals and 60 anticoagulated patients stabilized over 3 months in a therapeutic International Normalized Ratio (INR) range between 2-3.5 were studied. A drop of capillary blood was obtained simultaneously from two different fingers of each patient and applied to the monitor's application zone. The mean INR of the patients' blood samples of the two monitors differed by 0.01 units (2.32+/-0.63 for Pro Time and 2.33+/-0.68 for CoaguChek). The percentage of simple concordance and the kappa index were 88.3 and 75.9%, respectively. The coefficient of correlation was 0.922. The mean difference (bias) between the monitors was 0.01. The portable PT monitors evaluated presented a high percentage of concordance in INR results. PMID- 14745429 TI - In vitro effects of interferon-gamma and tumor necrosis factor-alpha on CD34+ bone marrow progenitor cells from aplastic anemia patients and normal donors. AB - Acquired aplastic anemia is characterized by loss or dysfunction of hematopoietic stem and progenitor cells. The proinflammatory cytokines Tumor necrosis factor alpha (TNF-alpha) and interferon-gamma (IFN-gamma) may be responsible for the immune-mediated pathology observed in some patients. The CD34+ population of bone marrow mononuclear cells contains primitive cells responsible for hemopoiesis. We investigated the response of CD34+ cells from aplastic anemia patients to a combination of IFN-gamma and TNF-alpha, and compared them to cells from normal volunteer donors. This was to determine whether aplastic CD34+ cells are more sensitive than normal cells to IFN-gamma/TNF-alpha-mediated effects, and whether cytokine-induced CD95 expression can explain the high levels of apoptosis observed in CD34+ cells from aplastic patients. CD34+38- cells were most affected by overnight incubation with these cytokines, their proportion and numbers being reduced in both normal donors and patients. There was no evidence for increased apoptosis, suggesting that this effect may be due to differentiation. IFN gamma/TNF-alpha induced upregulation of CD95 on both normal and aplastic CD34+ cells, although the basal level of CD95 expression was increased in aplastic cells. However, CD95 induction did not make cells from normal donors or aplastic anemia patients susceptible to induction of apoptosis by agonistic anti-CD95 antibodies, soluble CD95 ligand, or membrane-bound CD95L. In vivo CD95L is required for CD95 induced apoptosis. No forms of this protein were detectable in lymphocytes from aplastic patients. We conclude that increased apoptosis in aplastic CD34+ cells is not due to increased sensitivity to IFN-gamma/TNF-alpha. We further show that normal and aplastic CD34+ cells are resistant to CD95 apoptosis, even in the presence of mCD95L. PMID- 14745430 TI - Characterisation of TP53 abnormalities in chronic lymphocytic leukaemia. AB - Abnormalities of TP53 in chronic lymphocytic leukaemia (CLL) correlate with aggressive disease and transformation. We studied 115 patients with CLL including 90 untreated, 25 with heavily pretreated/refractory CLL using fluorescent in situ hybridisation (FISH) to detect allelic loss at chromosome 17p and flow cytometry (FC) to test p53 protein overexpression. A total of 17 cases were identified with TP53 deletion and/or protein expression. Both tests correlated in 10 of 17 patients; in six, one or the other abnormality was detected and in one case, with a deletion, flow cytometry failed. Material for direct DNA sequencing was available in 14 of 17 cases. Mutations were found in seven cases. Five of 14 patients with allelic loss and seven of 13 expressing p53 protein had a mutation. These were single-base substitutions and were located in exons 5, 7 or 8. Mutations were not found in 13 of 14 other cases without deletions by FISH or protein expression. The incidence of p53 abnormalities in this series was 15%, with a significant difference between untreated patients (7%) and the pretreated/refractory group (50%; P<0.01). Abnormal p53 was predicted for shorter survival, regardless of the method used. We confirm that p53 abnormalities are more common in refractory CLL and that mutations occur at the known hot spots. Testing for TP53 deletions by FISH and protein expression by FC is an effective and simple way of screening patients who are likely to have aggressive disease. DNA sequencing adds little to these methods in identifying the population at risk. PMID- 14745431 TI - High incidence of BCR-ABL kinase domain mutations and absence of mutations of the PDGFR and KIT activation loops in CML patients with secondary resistance to imatinib. AB - Imatinib, a specific inhibitor of the Abl, Kit and platelet-derived growth factor receptor (PDGFR) tyrosine kinases, is effective in all phases of chronic myelogenous leukemia. While responses in chronic phase are usually durable, resistance frequently develops in patients with advanced disease after an initial response. Several mechanisms of resistance have been demonstrated in vivo, including mutations in the BCR-ABL kinase domain and amplification of the BCR-ABL gene. We analyzed cytogenetics and screened for mutations of the BCR-ABL kinase domain as well as the activation loops of KIT and PDGFRA and B in 49 patients with CML or Ph-positive acute lymphoblastic leukemia with resistance to imatinib. Mutations in the kinase domain of BCR-ABL were detected in 51.6% of patients with secondary resistance but not in patients with primary resistance. Three of these mutations have not been described before (T315D, F359D and D276G). By contrast, KIT and PDGFRA and B were consistently wildtype. Clonal evolution prior to imatinib was present in 68.8% of patients with primary resistance and in 45.5% with secondary resistance. Additional cytogenetic aberrations developed in 18.2% of patients at the time of relapse. Our results confirm the high frequency of BCR ABL kinase domain mutations in patients with secondary resistance to imatinib and exclude mutations of the activation loops of KIT, PDGFRA and PDGFRB as possible causes of resistance in patients without ABL mutations. PMID- 14745432 TI - Use of polymorphisms in the noncoding region of the human mitochondrial genome to identify potential contamination of human leukemia-lymphoma cell lines. AB - The availability of the complete sequence of human mitochondrial DNA (mtDNA) has proven extremely useful in phylogenetic studies, forensic science and the determination of chimerism after allogeneic stem cell transplantation. In this study, we could demonstrate that the analysis of mtDNA polymorphisms is a quick and reliable method to identify contamination of human hematopoietic cell lines. This assay is based on PCR-sequencing of three hypervariable segments of the control region of mtDNA (hypervariable region (HRV) 1, 2 and 3). All three regions contain a large number of single-base polymorphisms. mtDNA was isolated according to standard laboratory procedures and amplified by PCR. Subsequently products were sequenced and evaluated with a semiautomated DNA sequencer system. So far, 21 human leukemia-lymphoma (LL) cell lines and nine other human cell lines were screened for contamination by other cell lines applying this method. We conclude that analysis of mtDNA polymorphisms is a quick, reliable and inexpensive method to detect intra - and interspecies cross-contamination and for the authentication of human LL cell lines. In comparison to other methods (cytogenetics, fluorescence in situ hybridization or immunophenotyping), this technique is less laborious and time consuming. PMID- 14745433 TI - A prospective comparison of fine-needle aspiration cytology and histopathology in the diagnosis and classification of lymphomas. AB - INTRODUCTION: Surgical biopsy examination is the gold standard for lymphoma diagnostics. However, fine-needle aspiration cytology (FNAC) offers several advantages in that it is quick, inexpensive, and the aspiration procedure has very few complications. This prospective study compares the diagnostic outcome between FNAC and surgical biopsy. MATERIALS AND METHODS: A total of 103 patients (>15 years) with lymphadenopathy and accessible lymph nodes underwent both diagnostic procedures. Immunophenotyping was performed on both FNAC and histopathological specimens. The updated KIEL classification was used for primary diagnosis and the WHO classification for reclassification. RESULTS: FNAC- and histopathology-based diagnoses were concordant in 76 patients. In 10 patients, there was a major diagnostic discordance: four differed with regard to degree of malignancy (low- versus high-grade NHL), three lymphoma versus reactive changes, and three regarding Hodgkin's lymphoma versus anaplastic large cell lymphoma. In 10 patients there was some (minor) discordance regarding subclassification: in eight patients the results of immunophenotyping differed, in two cases there were discrepancies in the cell type classification. In the remaining seven cases, there were diagnostic difficulties due to an insufficient sample. two serious adverse events occurred following surgical biopsy. CONCLUSIONS: FNAC is an accurate method in the diagnosis of lymphomas when the cytologic diagnosis is corroborated by immunophenotyping. However, an increasing use of FNAC for primary diagnosis and classification of lymphomas may result in a loss of archival tissue for complementary analyses, reclassification, and research purposes. In addition, some of the lymphoma entities are impossible to diagnose with use of the FNAC technique. PMID- 14745434 TI - Overwhelming postsplenectomy infection: is quality of patient knowledge enough for prevention? AB - BACKGROUND/OBJECTIVE: Fulminant, potentially life-threatening infection represents a major long-term risk after splenectomy. This study examines the impact of patient's knowledge and compliance on the prevention of overwhelming postsplenectomy infection (OPSI). METHODS: A Total of 318 splenectomized patients (median age: 18 years (10-26 years); M : F, 187 : 131) were enrolled in this study. A questionnaire was administered to assess the degree of knowledge and patient compliance and their role in the prevention of postsplenectomy risks; while identifying the group of health-care providers most successful in conveying information. RESULTS: The 318 patients had been splenectomized and followed up through a 17-year period. OPSI occurred among 5.7% (n=18) of patients. Of these, 56% occurred within the first 6 months and 44% in the following 10 years post splenectomy. Three patients died of OPSI, two during the first 6 months and one 2 years later. Of the followed up patients, 44.8% (n=142) had good knowledge of the risks of splenectomy and their prevention, 30.4% (n=96) had fair knowledge and 24.8% (n=79) had poor knowledge. Patients displaying greatest knowledge had a prevalence of OPSI of 1.4% compared to 16.5% among those with poor knowledge (P<0.001). In all, 60% of patients with good knowledge got their information principally from their tending hematologist. Among patients on regular and irregular prophylactic oral penicillin, OPSI occurred in 2.7 and 10% respectively (P<0.01). The incidence of OPSI also decreased from 7.3 to 3.2% after routine administration of pneumococcal vaccine (P<0.05). CONCLUSION: Although good knowledge, prophylactic penicillin and pneumococcal vaccination have remarkably reduced OPSI, it was not enough for complete prevention. The use of lifelong antibiotic prophylaxis remains of disputed value since no OPSI was recorded more than 10 years post splenectomy. PMID- 14745435 TI - Fludarabine-induced hemolytic anemia: successful treatment by rituximab. AB - Autoimmune disorders are frequent in chronic lymphocytic leukemia (CLL). Some drugs such as fludarabine may increase the risk of these phenomena and may even be fatal. The management of autoimmune disorders is not standard and corticosteroid is the most frequently used drug in these situations. Here, a case with CLL and severe autoimmune hemolytic anemia associated with fludarabine and successful treatment by rituximab has been reported. PMID- 14745436 TI - Acute myeloid leukemia with t(8;21)(q22;q22) manifesting as granulocytic sarcomas in the rhinopharynx and external acoustic meatus at relapse after high-dose cytarabine: case report and review of the literature. AB - We report a 31-year-old female with t(8;21)(q22;q22) acute myeloid leukemia (AML), M2 in the FAB classification. Complete remission was achieved with daunorubicin and cytarabine induction therapy followed by three courses of high dose cytarabine consolidation. Only 3 months later, the patient relapsed with granulocytic sarcomas (GSs) in her rhinopharynx, external acoustic meatus, and bone marrow. She received focal radiation for the GSs and successfully underwent reinduction chemotherapy. Subsequently, she received a matched related donor peripheral blood stem cell transplantation followed by high-dose chemotherapy and is now in a second remission. We summarized 79 reported cases of t(8;21) AML with GS and reviewed the literature to identify differences in the characteristics of t(8;21) AML with GS between adults and children. To our knowledge, this is the first report of pharyngeal GS in t(8;21) AML, and focal irradiation plus more intensive postinduction therapy during first remission, such as allogeneic-SCT, may be effective in adult t(8;21) AML patients with GS. PMID- 14745437 TI - Recurrent psychotic episodes in a patient who received rituximab for the treatment of non-Hodgkin's lymphoma. PMID- 14745438 TI - Myelodysplastic syndrome in a patient with hereditary pyruvate kinase deficiency. PMID- 14745439 TI - Synergistic effect of vitamin E and selenium in human prostate cancer cell lines. AB - Vitamin E and selenium are the two most popular dietary supplements used to prevent prostate cancer. The hypothesis that these antioxidants reduce prostate risk is being tested in the selenium and vitamin E chemoprevention trial (SELECT). We hypothesize that selenium potentiates vitamin E-induced inhibition of prostate cancer cell growth in vitro. Prostate cancer cell populations growing asynchronously were treated with a combination of vitamin E and selenium and processed for flow cytometric analysis. Prostate cancer cells treated with a combination of the antioxidants revealed that selenium potentiates vitamin E induced inhibition of LNCaP cells in vitro. This was demonstrated by a reduction in the percentage of cells in the S phase. This crucial finding confirms our previous observations that antioxidant molecules act via distinct mechanistic pathways. These independent biological effects can be exploited in order to augment the anticancer properties of individual agents. These data also validate the two factorial design of the SELECT trial, permitting pairwise comparisons between agents in combination and alone. PMID- 14745440 TI - Solution structure of domain 5 of a group II intron ribozyme reveals a new RNA motif. AB - Domain 5 (D5) is the central core of group II intron ribozymes. Many base and backbone substituents of this highly conserved hairpin participate in catalysis and are crucial for binding to other intron domains. We report the solution structures of the 34-nucleotide D5 hairpin from the group II intron ai5 gamma in the absence and presence of divalent metal ions. The bulge region of D5 adopts a novel fold, where G26 adopts a syn conformation and flips down into the major groove of helix 1, close to the major groove face of the catalytic AGC triad. The backbone near G26 is kinked, exposing the base plane of the adjacent A-U pair to the solvent and causing bases of the bulge to stack intercalatively. Metal ion titrations reveal strong Mg(2+) binding to a minor groove shelf in the D5 bulge. Another distinct metal ion-binding site is observed along the minor groove side of the catalytic triad, in a manner consistent with metal ion binding in the ribozyme active site. PMID- 14745441 TI - Reactive oxygen species impair Slo1 BK channel function by altering cysteine mediated calcium sensing. AB - Vascular dysfunction is a hallmark of many diseases, including coronary heart disease, stroke and diabetes. The underlying mechanisms of these disorders, which are intimately associated with inflammation and oxidative stress caused by excess reactive oxygen species (ROS), have remained elusive. Here we report that ROS are powerful inhibitors of vascular smooth muscle calcium-dependent Slo1 BK or Maxi-K potassium channels, an important physiological determinant of vascular tone. By targeting a cysteine residue near the Ca(2+) bowl of the BK alpha subunit, H(2)O(2) virtually eliminates physiological activation of the channel, with an inhibitory potency comparable to a knockout of the auxiliary subunit BK beta 1. These results reveal a molecular structural basis for the vascular dysfunction involving oxidative stress and provide a solid rationale for a potential use of BK openers in the prevention and treatment of cardiovascular disorders. PMID- 14745442 TI - Differentiating the pathologies of cerebral malaria by postmortem parasite counts. AB - To study the pathogenesis of fatal cerebral malaria, we conducted autopsies in 31 children with this clinical diagnosis. We found that 23% of the children had actually died from other causes. The remaining patients had parasites sequestered in cerebral capillaries, and 75% of those had additional intra- and perivascular pathology. Retinopathy was the only clinical sign distinguishing malarial from nonmalarial coma. These data have implications for treating malaria patients, designing clinical trials and assessing malaria-specific disease associations. PMID- 14745443 TI - Lymphoid follicle destruction and immunosuppression after repeated CpG oligodeoxynucleotide administration. AB - DNA containing unmethylated cytidyl guanosyl (CpG) sequences, which are underrepresented in mammalian genomes but prevalent in prokaryotes, is endocytosed by cells of the innate immune system, including macrophages, monocytes and dendritic cells, and activates a pathway involving Toll-like receptor-9 (TLR9). CpG-containing oligodeoxynucleotides (CpG-ODN) are potent stimulators of innate immunity, and are currently being tested as adjuvants of antimicrobial, antiallergic, anticancer and antiprion immunotherapy. Little is known, however, about the consequences of repeated CpG-ODN administration, which is advocated for some of these applications. Here we report that daily injection of 60 microg CpG-ODN dramatically alters the morphology and functionality of mouse lymphoid organs. By day 7, lymphoid follicles were poorly defined; follicular dendritic cells (FDC) and germinal center B lymphocytes were suppressed. Accordingly, CpG-ODN treatment for > or =7 d strongly reduced primary humoral immune responses and immunoglobulin class switching. By day 20, mice developed multifocal liver necrosis and hemorrhagic ascites. All untoward effects were strictly dependent on CpG and TLR9, as neither the CpG-ODN treatment of Tlr9(-/-) mice nor the repetitive challenge of wild-type mice with nonstimulatory ODN (AT-ODN) or with the TLR3 agonist polyinosinic:cytidylic acid (polyI:C) were immunotoxic or hepatotoxic. PMID- 14745445 TI - Ligand-independent redistribution of Fas (CD95) into lipid rafts mediates clonotypic T cell death. AB - Clonotypic elimination of activated T cells through Fas-Fas ligand (CD95-CD95L) interactions is one mechanism of peripheral self-tolerance. T cell receptor (TCR) stimuli trigger FasL synthesis but also sensitize activated T cells to Fas mediated apoptosis through an unknown mechanism. Here we show that TCR restimulation of activated human CD4(+) T cells resulted in Fas translocation into lipid raft microdomains before binding FasL, rendering these cells sensitive to apoptosis after stimulation with bivalent antibody or FasL. Disruption of lipid rafts reduced sensitivity to Fas-mediated apoptosis after TCR restimulation. Thus, the redistribution of Fas and other tumor necrosis factor family receptors into and out of lipid rafts may dynamically regulate the efficiency and outcomes of signaling by these receptors. PMID- 14745446 TI - MHC class Ia-restricted memory T cells inhibit expansion of a nonprotective MHC class Ib (H2-M3)-restricted memory response. AB - Listeria monocytogenes infection generates major histocompatibility complex (MHC) class Ia-restricted and MHC class Ib-(H2-M3)-restricted effector and memory CD8+ T cells. However, only MHC class Ia-restricted memory cells expand after rechallenge, and it is unknown if MHC class Ib-restricted memory CD8+ T cells generated by vaccination are protective. We show here that H2-M3-restricted memory CD8+ T cells were capable of secondary expansion but, in contrast to primary H2-M3-restricted effector cells, failed to provide protective immunity. In lm-immune mice, MHC class Ia-restricted memory CD8+ T cells prevented the expansion of H2-M3-restricted memory T cell populations by limiting dendritic cell antigen presentation. Thus, protective immunity by H2-M3-restricted T cells is limited to primary infection, indicating that memory MHC class Ia-restricted T cells prevent nonessential immune responses during secondary infection. PMID- 14745444 TI - Direct evidence that the VEGF-specific antibody bevacizumab has antivascular effects in human rectal cancer. AB - The effects of vascular endothelial growth factor (VEGF) blockade on the vascular biology of human tumors are not known. Here we show here that a single infusion of the VEGF-specific antibody bevacizumab decreases tumor perfusion, vascular volume, microvascular density, interstitial fluid pressure and the number of viable, circulating endothelial and progenitor cells, and increases the fraction of vessels with pericyte coverage in rectal carcinoma patients. These data indicate that VEGF blockade has a direct and rapid antivascular effect in human tumors. PMID- 14745447 TI - Regulation of flowering time by FVE, a retinoblastoma-associated protein. AB - The initiation of flowering in plants is controlled by environmental and endogenous signals. Molecular analysis of this process in Arabidopsis thaliana indicates that environmental control is exerted through the photoperiod and vernalization pathways, whereas endogenous signals regulate the autonomous and gibberellin pathways. The vernalization and autonomous pathways converge on the negative regulation of FLC, a gene encoding a MADS-box protein that inhibits flowering. We cloned FVE, a component of the autonomous pathway that encodes AtMSI4, a putative retinoblastoma-associated protein. FVE interacted with retinoblastoma protein in immunoprecipitation assays, and FLC chromatin was enriched in acetylated histones in fve mutants. We conclude that FVE participates in a protein complex repressing FLC transcription through a histone deacetylation mechanism. Our data provide genetic evidence of a new developmental function of these conserved proteins and identify a new genetic mechanism in the regulation of flowering. PMID- 14745448 TI - Convergent evidence for impaired AKT1-GSK3beta signaling in schizophrenia. AB - AKT-GSK3beta signaling is a target of lithium and as such has been implicated in the pathogenesis of mood disorders. Here, we provide evidence that this signaling pathway also has a role in schizophrenia. Specifically, we present convergent evidence for a decrease in AKT1 protein levels and levels of phosphorylation of GSK3beta at Ser9 in the peripheral lymphocytes and brains of individuals with schizophrenia; a significant association between schizophrenia and an AKT1 haplotype associated with lower AKT1 protein levels; and a greater sensitivity to the sensorimotor gating-disruptive effect of amphetamine, conferred by AKT1 deficiency. Our findings support the proposal that alterations in AKT1-GSK3beta signaling contribute to schizophrenia pathogenesis and identify AKT1 as a potential schizophrenia susceptibility gene. Consistent with this proposal, we also show that haloperidol induces a stepwise increase in regulatory phosphorylation of AKT1 in the brains of treated mice that could compensate for an impaired function of this signaling pathway in schizophrenia. PMID- 14745449 TI - Elastic fiber homeostasis requires lysyl oxidase-like 1 protein. AB - Elastic fibers are components of the extracellular matrix and confer resilience. Once laid down, they are thought to remain stable, except in the uterine tract where cycles of active remodeling occur. Loss of elastic fibers underlies connective tissue aging and important diseases including emphysema. Failure to maintain elastic fibers is explained by a theory of antielastase-elastase imbalance, but little is known about the role of renewal. Here we show that mice lacking the protein lysyl oxidase-like 1 (LOXL1) do not deposit normal elastic fibers in the uterine tract post partum and develop pelvic organ prolapse, enlarged airspaces of the lung, loose skin and vascular abnormalities with concomitant tropoelastin accumulation. Distinct from the prototypic lysyl oxidase (LOX), LOXL1 localizes specifically to sites of elastogenesis and interacts with fibulin-5. Thus elastin polymer deposition is a crucial aspect of elastic fiber maintenance and is dependent on LOXL1, which serves both as a cross-linking enzyme and an element of the scaffold to ensure spatially defined deposition of elastin. PMID- 14745450 TI - A genetic link between cold responses and flowering time through FVE in Arabidopsis thaliana. AB - Cold induces expression of a number of genes that encode proteins that enhance tolerance to freezing temperatures in plants. A cis-acting element responsive to cold and drought, the C-repeat/dehydration-responsive element (C/DRE), was identified in the Arabidopsis thaliana stress-inducible genes RD29A and COR15a and found in other cold-inducible genes in various plants. C/DRE-binding factor/DRE-binding protein (CBF/DREB) is an essential component of the cold acclimation response, but the signaling pathways and networks are mostly unknown. Here we used targeted genetic approach to isolate A. thaliana mutants with altered cold-responsive gene expression (acg) and identify ACG1 as a negative regulator of the CBF/DREB pathway. acg1 flowered late and had elevated expression of FLOWERING LOCUS C (FLC), a repressor of flowering encoding a MADS-box protein. We showed that acg1 is a null allele of the autonomous pathway gene FVE. FVE encodes a homolog of the mammalian retinoblastoma-associated protein, a component of a histone deacetylase (HDAC) complex involved in transcriptional repression. We also showed that plants sense intermittent cold stress through FVE and delay flowering with increasing expression of FLC. Dual roles of FVE in regulating the flowering time and the cold response may have an evolutionary advantage for plants by increasing their survival rates. PMID- 14745451 TI - Bi-directional effects of GABA(B) receptor agonists on the mesolimbic dopamine system. AB - The rewarding effect of drugs of abuse is mediated by activation of the mesolimbic dopamine system, which is inhibited by putative anti-craving compounds. Interestingly, different GABA(B) receptor agonists can exert similarly opposing effects on the reward pathway, but the cellular mechanisms involved are unknown. Here we found that the coupling efficacy (EC(50)) of G-protein-gated inwardly rectifying potassium (GIRK, Kir3) channels to GABA(B) receptor was much lower in dopamine neurons than in GABA neurons of the ventral tegmental area (VTA), depending on the differential expression of GIRK subunits. Consequently, in rodent VTA slices, a low concentration of the canonical agonist baclofen caused increased activity, whereas higher doses eventually inhibited dopamine neurons. At behaviorally relevant dosages, baclofen activated GIRK channels in both cell types, but the drug of abuse gamma-hydroxy-butyric acid (GHB) activated GIRK channels only in GABAergic neurons. Thus GABA(B) receptor agonists exert parallel cellular and behavioral effects due to the cell-specific expression of GIRK subunits. PMID- 14745452 TI - Random presentation enables subjects to adapt to two opposing forces on the hand. AB - Studies have shown that humans cannot simultaneously learn opposing force fields or opposing visuomotor rotations, even when provided with arbitrary contextual information, probably because of interference in their working memory. In contrast, we found that subjects can adapt to two opposing force fields when provided with contextual cues and can consolidate motor memories if random and frequent switching occurs. Because significant aftereffects were seen, this study suggests that multiple internal models can be acquired simultaneously during learning and predictively switched, depending only on contextual information. PMID- 14745453 TI - Diclofenac residues as the cause of vulture population decline in Pakistan. AB - The Oriental white-backed vulture (OWBV; Gyps bengalensis) was once one of the most common raptors in the Indian subcontinent. A population decline of >95%, starting in the 1990s, was first noted at Keoladeo National Park, India. Since then, catastrophic declines, also involving Gyps indicus and Gyps tenuirostris, have continued to be reported across the subcontinent. Consequently these vultures are now listed as critically endangered by BirdLife International. In 2000, the Peregrine Fund initiated its Asian Vulture Crisis Project with the Ornithological Society of Pakistan, establishing study sites at 16 OWBV colonies in the Kasur, Khanewal and Muzaffargarh-Layyah Districts of Pakistan to measure mortality at over 2,400 active nest sites. Between 2000 and 2003, high annual adult and subadult mortality (5-86%) and resulting population declines (34-95%) (ref. 5 and M.G., manuscript in preparation) were associated with renal failure and visceral gout. Here, we provide results that directly correlate residues of the anti-inflammatory drug diclofenac with renal failure. Diclofenac residues and renal disease were reproduced experimentally in OWBVs by direct oral exposure and through feeding vultures diclofenac-treated livestock. We propose that residues of veterinary diclofenac are responsible for the OWBV decline. PMID- 14745454 TI - Association study of an SNP combination pattern in the dopaminergic pathway in paranoid schizophrenia: a novel strategy for complex disorders. AB - Schizophrenia is a common mental disorder with a complex pattern of inheritance. Despite a large number of studies in the past decades, its molecular etiology remains unknown. In this study, we proposed a 'system-thinking' strategy in seeking the combined effect of susceptibility genes for a complex disorder by using paranoid schizophrenia as an example. We genotyped 85 reported single nucleotide polymorphisms (SNPs) present in 23 genes for the dopamine (DA) metabolism pathway among 83 paranoid schizophrenics and 108 normal controls with detailed clinical and genetic information. We developed two novel multilocus approaches-the potential effective SNP combination pattern and potential effective dynamic effects analysis, by which three susceptibility genotype combinations were found to be associated with schizophrenia. These results were also validated in a family-based cohort consisting of 95 family trios of paranoid schizophrenia. The present findings suggest that the COMT and ALDH3 combination may be the most common type involved in predisposing to schizophrenia. Since the combination blocks the whole pathways for the breakdown of DA and noradrenaline, it is very likely to play a central role in developing paranoid schizophrenia. PMID- 14745457 TI - Impairment of hepatic microcirculation in fatty liver. AB - Fatty liver or hepatic steatosis, which is the result of the abnormal accumulation of triacylglycerol within the cytoplasm of hepatocytes, is a common histological finding in human liver biopsy specimens that is attributed to the effects of alcohol excess, obesity, diabetes, or drugs. There is a general consensus that fatty liver compromises hepatic microcirculation, the common exchange network upon which hepatic arterial and portal inflows converge, regardless of underlying etiology. A significant reduction in hepatic microcirculation has been observed in human fatty donor livers and in experimental models of hepatic steatosis. There is an inverse correlation between the degree of fat infiltration and both total hepatic blood flow and flow in microcirculation. Fatty accumulation in the cytoplasm of the hepatocytes is associated with an increase in the cell volume that reduces the size of the hepatic sinusoid space by 50% compared with a normal liver and may result in partial or complete obstruction of the hepatic sinusoid space. As a result of impaired hepatic microcirculation, the hepatocytes of the fatty liver have reduced tolerance against ischemia-reperfusion injury, which affects about 25% of the donors for liver transplantation because severe steatosis is associated with a high risk of primary nonfunction after liver transplantation. PMID- 14745455 TI - Activation of methionine synthase by insulin-like growth factor-1 and dopamine: a target for neurodevelopmental toxins and thimerosal. AB - Methylation events play a critical role in the ability of growth factors to promote normal development. Neurodevelopmental toxins, such as ethanol and heavy metals, interrupt growth factor signaling, raising the possibility that they might exert adverse effects on methylation. We found that insulin-like growth factor-1 (IGF-1)- and dopamine-stimulated methionine synthase (MS) activity and folate-dependent methylation of phospholipids in SH-SY5Y human neuroblastoma cells, via a PI3-kinase- and MAP-kinase-dependent mechanism. The stimulation of this pathway increased DNA methylation, while its inhibition increased methylation-sensitive gene expression. Ethanol potently interfered with IGF-1 activation of MS and blocked its effect on DNA methylation, whereas it did not inhibit the effects of dopamine. Metal ions potently affected IGF-1 and dopamine stimulated MS activity, as well as folate-dependent phospholipid methylation: Cu(2+) promoted enzyme activity and methylation, while Cu(+), Pb(2+), Hg(2+) and Al(3+) were inhibitory. The ethylmercury-containing preservative thimerosal inhibited both IGF-1- and dopamine-stimulated methylation with an IC(50) of 1 nM and eliminated MS activity. Our findings outline a novel growth factor signaling pathway that regulates MS activity and thereby modulates methylation reactions, including DNA methylation. The potent inhibition of this pathway by ethanol, lead, mercury, aluminum and thimerosal suggests that it may be an important target of neurodevelopmental toxins. PMID- 14745458 TI - Identification of a novel route of iron transcytosis across the mammalian blood brain barrier. AB - OBJECTIVE: This study was undertaken to assess the role of p97 (also known as melanotransferrin) in the transfer of iron into the brain, because the passage of most large molecules is limited by the presence of the blood-brain barrier, including that of the serum iron transporter transferrin. METHODS: To study the function of the soluble form of p97, we followed the uptake of radioiodinated and 55Fe loaded p97 and transferrin by the brain during a 24-hour period. RESULTS: We show that the soluble form of p97 has the ability to transcytose across the murine blood-brain barrier, and its transcytosis can be inhibited in a specific manner. We also provide evidence that p97 transports iron into the brain more efficiently than transferrin. CONCLUSIONS: These data support the idea that p97 is an important iron transporter across the blood-brain barrier in normal physiology and possibly in neurodegenerative diseases, such as Alzheimer disease, in which iron homeostasis in the brain becomes disrupted. PMID- 14745459 TI - Blood-brain barrier permeability precedes senile plaque formation in an Alzheimer disease model. AB - OBJECTIVE: To establish the generality of cerebrovascular pathology frequently observed with Alzheimer disease, we have assessed blood-brain barrier (BBB) integrity using the Alzheimer disease model Tg2576 mice in which cognitive deficits and neuritic plaque formation develop around 10-12 months of age. METHODS: We assessed BBB integrity using well-established methods involving albumin and Evans blue uptake and introduce the use of a novel perfusion protocol using succinimidyl ester of carboxyfluorescein diacetate. RESULTS: BBB permeability is increased in the cerebral cortex of 10-month-old Tg2576 mice preceding Alzheimer disease pathology presentation. Furthermore, when compared with their nontransgenic littermates, 4-month-old Tg2576 mice exhibit compromised BBB integrity in some areas of the cerebral cortex. An age-related increase in albumin uptake by the brains of Tg2576 mice, compared with nontransgenic mice, was also observed. These findings were supported by quantitative Evans blue analysis (p = 0.07, two-way analysis of variance). CONCLUSION: A breakdown of BBB was evident in young 4- to 10-month-old Tg2576 mice. Compromised barrier function could explain the mechanisms of Abeta entry into the brain observed in experimental Alzheimer disease vaccination models. Such structural changes to the BBB caused by elevated Abeta could play a central role in Alzheimer disease development and might define an early point of intervention for designing effective therapy against the disease. PMID- 14745460 TI - Microvascular abnormalities in chronic heart failure: a cross-sectional analysis. AB - OBJECTIVES: Similar to what has been found in hypertension, elevated peripheral resistance in chronic heart failure (CHF) might be related to microvascular constriction and rarefaction. Our objective was to evaluate both structural and functional microvascular changes in patients with CHF in relation to left ventricular function and neurohumoral activation. METHODS: In 25 patients with mild and severe CHF (New York Heart Association class I-II [n = 11] and class III IV [n = 14]) and 10 age-matched healthy subjects, we studied microvascular density, diameters, and morphology of the bulbar conjunctiva and skin nailfold using intravital microscopy. RESULTS: Total conjunctival microvascular density was higher in patients with mild heart failure compared with healthy controls, whereas it was lower in severe heart failure (medians, 6.75, 4.31, and 3.56 mm/mm2, respectively, p < 0.01). In patients with heart failure, venular density was correlated with left ventricular ejection fraction. Nailfold capillary recruitment during postocclusive reactive hyperemia, a measure of functional reserve capacity, was impaired in patients with severe CHF (p < 0.05). Furthermore, in severe CHF, more abnormal capillaries and enlarged diameters were found in the nailfold (p < 0.05). CONCLUSIONS: In heart failure, several microvascular abnormalities occur that differ, depending on the severity of this condition. PMID- 14745461 TI - A model of nitric oxide capillary exchange. AB - OBJECTIVE: Our aim was to develop a mathematical model that describes the nitric oxide (NO) transport in and around capillaries. The model is used to make quantitative predictions for (1) the contribution of capillary endothelium to the nitric oxide flux into the parenchymal tissue cells; (2) the scavenging of arteriolar endothelium-derived NO by capillaries in the surrounding tissue; and (3) the role of myoglobin in tissue cells and plasma-based hemoglobin on NO diffusion in and around capillaries. METHODS: We used a finite element model of a capillary and surrounding tissue with discrete parachute-shape red blood cells (RBCs) moving inside the capillary to obtain the NO concentration distribution. An intravascular mass transfer coefficient is estimated as a function of RBC membrane permeability and capillary hematocrit. A continuum model of the capillary is also formulated, in which blood is treated as a homogeneous fluid; it uses the mass transfer coefficient and provides a closed-form analytic solution for the average exchange rate of NO in a capillary-perfused region. RESULTS: The NO concentration in the parenchymal cells depends on parameters such as RBC membrane permeability and capillary hematocrit; the concentration is predicted for a wide range of parameters. In the absence of myoglobin or plasma based hemoglobin, the average tissue concentration generally ranges between 20 and 300 nM. In the presence of myoglobin or after transfusion of a hemoglobin based blood substitute, there is minimal NO penetration into the tissue from the capillary endothelium. CONCLUSIONS: The model suggests that NO originating from the capillary wall can diffuse toward the parenchymal cells and potentially sustain physiologically significant concentrations. The model provides estimates of NO exchange and concentration level in capillary-perfused tissue, and it can be used in models of NO transport around arterioles or other NO sources. PMID- 14745463 TI - Receptive field scatter, topography and map variability in different layers of the hindpaw representation of rat somatosensory cortex. AB - We recorded neurons extracellularly in layers II/III, IV, and V of the hindpaw representation of primary somatosensory cortex in anesthetized rats and studied laminar features of receptive fields (RFs) and representational maps. On average, RFs were smallest in layer IV and largest in layer V; however, for individual penetrations we found substantial deviations from this rule. Within the hindpaw representation, a distinct rostrocaudal gradient of RF size was present in all layers. While layer V RFs were generally largest independent of this gradient, layer IV RFs recorded caudally representing the proximal portions of the paw were larger than layer II/III RFs recorded rostrally representing the digits. The individual scatter of the locations of RFs across laminar groups was in the range of several millimeters, corresponding to about 25% of the average RF diameter. The cutaneous representations of the hindpaw in extragranular layers were confined to the areal extent defined by responsive sites in layer IV. Comparison between RFs determined quantitatively and by handplotting showed a reliable correspondence. Repeated measurements of RFs revealed spontaneous fluctuations of RF size of no more than 5% of the initial condition over an observation period of several hours. The topography and variability of cortical maps of the hindpaw representation were studied with a quantitative interpolation method taking into account the geometric centers of RFs and the corresponding cortical recording sites. On average, the overall topography in terms of preservation of neighborhood relations was present in all layers, although some individual maps showed severe distortions of topography. Factors contributing to map variability were overall position of the representation on the cortical surface, internal topography and spatial extent. Interindividual variability of map layout was always highest in the digit representations. Local topographic orderliness was lowest in layer V, but comparable in layers II/III and IV. Within layer IV, the lowest orderliness was observed in the digit representations. Our data emphasize a substantial variability of RF size, overlap and position across layers and within layers. At the level of representational maps, we found a similar degree of variability that often co-varied across layers, with little evidence for significant layer specificity. Laminar differences are likely to arise from the specific input-output pattern, layer-specific cell types and the connectivity between different layers. Our findings emphasizing similarities in the variability across layers support the notion of tightly coupled columnar interactions between different layers. PMID- 14745464 TI - Stimulation-induced inhibition of neuronal firing in human subthalamic nucleus. AB - The subthalamic nucleus (STN) is an important component of the basal ganglia (BG) and plays a major role in the pathogenesis of Parkinson's disease (PD). Hyperactivity of STN as a consequence of the loss of dopaminergic inputs to the BG is believed to be a major factor in producing the motor symptoms of PD. High frequency (HF) deep brain stimulation (DBS) of the STN has recently become an important treatment in PD patients where medications no longer provide satisfactory therapy. However, the mechanisms underlying DBS therapy are unknown, and there is seemingly conflicting data suggesting inhibition or excitation of STN neurons. This study directly examined the effects of stimulation in STN on the activity of STN neurons in PD patients during functional stereotactic mapping prior to insertion of DBS electrodes. Electrical stimulation in STN was investigated in twelve PD patients by recording the neural activity of a cell in STN with one electrode while applying current pulses through a second electrode located about 600 microm away. Stimulation at high frequencies (100-300 Hz) was found to produce inhibition following the stimulus train in 42% of the 60 cells tested. Inhibition during the train was seen in 13 of 15 neurons where it was possible to detect such activity. Furthermore, in 44% of the cases where HF stimulation produced inhibition there was an early inhibition followed by rebound excitation and a further inhibitory period, suggesting that the inhibitions observed are due to hyperpolarization. In eight of the 25 neurons inhibited by HF stimulation, the effects of single stimuli were determined and revealed that in seven of these there was an inhibitory period of 15-20 ms following each stimulus. Thus, the present findings suggest that local HF stimulation inhibits many STN neurons. However, these studies could not determine whether the stimulus also directly excited the cell and/or its axon, but other recent findings suggest that this is likely the case. Therefore, the overall effects of DBS stimulation in STN are likely to be inhibition of intrinsic and synaptically mediated activity, and its replacement by regular high-frequency firing. PMID- 14745465 TI - What drives children's limb selection for reaching in hemispace? AB - Arguably, the act of reaching constitutes one of the most devoted lines of contemporary developmental research. In addition to the underlying dynamical characteristics of motor coordination, a key element in programming is limb selection, a phenomenon (handedness) that has so far resisted any reasonable unified explanation. From a more contemporary view, two factors appear to have the most influence on hand selection for a given task: motor dominance and attentional information related to task demands. This study was designed to determine what factor(s) influence choice of limb for reaching in hemispace in reference to motor dominance, object proximity, and a hemispheric bias favoring use of the hand on the same side as the stimulus. Strong right-handed children were asked to reach and retrieve a small object across right and left hemispace locations beginning with the arms uncrossed and arms-crossed. With the arms crossed condition, an imagined and actual movement execution was administered. Results from the uncrossed condition supported previous reported findings for adults and children. That is, participants responded ipsilaterally using the hand on the same side as the stimulus, thus supporting the case for object proximity and hemispheric bias. However, in the arms-crossed condition the vast majority of participants preferred keeping the limbs crossed in response to right and left hemispace stimuli, which leads to the suggestion that object proximity rather than hemispheric bias was the driving factor in this context. The behavioral pattern for imagined and actual movement was not significantly different. Overall, the findings add to the growing acceptance that limb selection is task and context dependent, rather than a biologically based invariant feature of motor behavior. PMID- 14745466 TI - Use-dependent cortical plasticity in thalidomide-induced upper extremity dysplasia: evidence from somaesthesia and neuroimaging. AB - In this study cerebral reorganization was investigated in thalidomide-damaged subjects who use their feet to compensate for their malformed upper extremities. Tactile localization across toes was combined with fMRI to study use-dependent plasticity of the human somatosensory cortex. The manner of compensatory foot use was assessed by a questionnaire. In the behavioural experiment toes were stimulated with above threshold monofilaments and subjects had to report which toe was stimulated. When feet were employed for all everyday actions subjects made significantly fewer errors in the localization task. In subjects who use their feet only for specific actions such as grasping objects there were as many localization errors as in the control group of thalidomide-affected subjects with normal extremities. However, the patterns of mislocalizations were different with less errors occurring for the toe of the dominant foot involved in these actions. Functional MRI showed stronger haemodynamic responses to electrical stimulation of the toes in subjects using their feet for everyday actions as compared to controls. Our data show that long-term use of the feet for fine sensorimotor skills leads to better performance in tactile localization and changes in cerebral SI representation supporting the notion of use-dependent plasticity in the somatosensory cortex. PMID- 14745467 TI - Effects of transcranial direct current stimulation over the human motor cortex on corticospinal and transcallosal excitability. AB - Weak transcranial direct current stimulation (tDCS) can induce long lasting changes in cortical excitability. In the present study we asked whether tDCS applied to the left primary motor cortex (M1) also produces aftereffects distant from the site of the stimulating electrodes. We therefore tested corticospinal excitability in the left and the right M1 and transcallosal excitability between the two cortices using transcranial magnetic stimulation (TMS) before and after applying tDCS. Eight healthy subjects received 10 min of anodal or cathodal tDCS (1 mA) to the left M1. We examined the amplitude of contralateral motor evoked potentials (MEPs) and the onset latency and duration of transcallosal inhibition with single pulse TMS. MEPs evoked from the tDCS stimulated (left) M1 were increased by 32% after anodal and decreased by 27% after cathodal tDCS, while transcallosal inhibition evoked from the left M1 remained unchanged. The effect on MEPs evoked from the left M1 lasted longer for cathodal than for anodal tDCS. MEPs evoked from the right M1 were unchanged whilst the duration of transcallosal inhibition evoked from the right M1 was shortened after cathodal tDCS and prolonged after anodal tDCS. The duration of transcallosal inhibition returned to control values before the effect on the MEPs from the left M1 had recovered. These findings are compatible with the idea that tDCS-induced aftereffects in the cortical motor system are limited to the stimulated hemisphere, and that tDCS not only affects corticospinal circuits involved in producing MEPs but also inhibitory interneurons mediating transcallosal inhibition from the contralateral hemisphere. PMID- 14745468 TI - Unconscious modulation of motor cortex excitability revealed with transcranial magnetic stimulation. AB - The neuronal effects of sensory events that do not enter conscious awareness have been reported in numerous pathological conditions and in normal subjects. In the present study, unconscious modulation of corticospinal excitability was probed in healthy volunteers with transcranial magnetic stimulation (TMS). TMS-induced motor evoked potentials (MEPs) were collected from the first dorsal interosseus muscle while subjects performed a masked semantic priming task that has been shown to elicit covert motor cortex activations. Our data show that the amplitude of the MEPs is modulated by an unseen prime, in line with temporal patterns revealed with event related potentials. These data confirm previous reports showing specific motor neural responses associated with an unseen visual stimulus and establish TMS as a valuable tool in the study of the neural correlates of consciousness. PMID- 14745469 TI - Anionic liposomes in capillary electrophoresis: effect of calcium on 1-palmitoyl 2-oleyl- sn-glycero-3-phosphatidylcholine / phosphatidylserine-coating in silica capillaries. AB - The effect of calcium on phospholipid coatings in fused silica capillaries used in capillary electrophoresis was studied. The anionic liposomes used for the coating consisted of 3 mM 1-palmitoyl-2-oleyl- sn-glycero-3-phosphatidylcholine and phosphatidylserine in the ratio 80/20 mol%. Coating was performed as part of the preconditioning, and the capillaries could be used for several runs without the need for liposomes in the background electrolyte solution or for liposome rinses between runs. Phospholipids could easily be flushed away by rinsing with a chloroform-methanol (2:1 v/v) mixture, which made it possible to recoat and reuse the capillaries. A calcium:phospholipid ratio of approximately 3 gave the most stable coating. The stability of the coating and success of the coating procedure were studied by measuring the electroosmotic flow and by separating uncharged steroids, which were used as model compounds. Many parameters that affect the coating, such as preconditioning (with different acids and bases), buffer, temperature during coating, and the physical structures of liposomes, were studied, with and without calcium in the liposome solution. The separation of steroids was improved and was less dependent on coating conditions when calcium was present during the coating. Capillaries optimally coated with anionic phospholipids were applied in the separation of phenols. PMID- 14745470 TI - Preparation and certification of a reference material for the determination of nutrients in seawater. AB - There is an urgent need for natural water reference materials certified for nutrients. In 1996, NRC collected seawater for a proposed CRM at a depth of 200 m in the North Atlantic; this was immediately filtered through 0.05-microm cartridge filters into 50-L carboys. The water was later homogenized in the NRC laboratories in Ottawa and stabilized via gamma irradiation. Over six years of stability testing no significant deterioration was detected. In addition to the usual customary standard colorimetric procedures, alternative analytical methods were developed to enable the certification process. The production of a CRM called MOOS-1 will be discussed. Certified values, with uncertainty components addressing the homogeneity, stability, and characterization of the material, were calculated to be: orthophosphate=1.56+/-0.07 micro mol L(-1), silicate=26.0+/-1.0 micro mol L(-1), nitrite=3.06+/-0.15 micro mol L(-1), and nitrite and nitrate=23.7+/-0.9 micro mol L(-1). PMID- 14745471 TI - Determination of vanadium in mussels by electrothermal atomic absorption spectrometry without chemical modifiers. AB - A method was developed for the quantitative determination of total vanadium concentration in mussels via electrothermal atomic absorption spectrometry (ETAAS). After the microwave digestion of the samples, a program using temperatures of 1600 degrees C and 2600 degrees C for ashing and atomization respectively, without any matrix modifiers, allowed us to obtain results that were satisfactory since they agreed closely with certified reference material values. The detection limit was 0.03 mg kg(-1) (dry weight), indicating that the method is suitable for the analysis of mussel samples. This determination was compared with matrix modifiers that have been reported previously. The method was applied to various cultivated and wild mussels from the Galician coast, yielding levels below 1 mg kg(-1) (wet weight). PMID- 14745473 TI - Biomimetic piezoelectric quartz sensor for caffeine based on a molecularly imprinted polymer. AB - A piezoelectric quartz sensor coated with molecularly imprinted polymer (MIP) for caffeine was developed. The MIP was prepared by co-polymerizing methacrylic acid (MAA) and ethylene glycol dimethacrylate (EDMA) in the presence of azobis(isobutyronitrile) as initiator, caffeine as template molecule, and chloroform as solvent. The MIP suspension in polyvinyl chloride/tetrahydrofuran (6:2:1 w/w/v) solution was spin coated onto the surface of the electrode of a 10 MHz AT-cut quartz crystal. The sensor exhibited a linear relationship between the frequency shift and caffeine concentration in the range of 1 x 10(-7) mg mL(-1 )up to 1 x 10(-3) mg mL(-1) [correlation coefficient ( r)=0.9935] in a stopped flow measurement mode. It has a sensitivity of about 24 Hz/ln(concentration, mg mL(-1)). A steady-state response was achieved in less than 10 min. The performance characteristic of the sensor shows a promising and inexpensive alternative method of detecting caffeine. Surface studies were carried out for the reagent phase of the sensor using SEM, AFM, and XPS analysis in order to elucidate the imprinting of the caffeine molecule. The SEM micrograph, AFM image, and XPS spectra confirmed the removal of caffeine by Soxhlet extraction in the imprinting process and the rebinding of caffeine to the MIP sensing layer during measurement. PMID- 14745474 TI - Dynamic ultrasound-assisted leaching of essential macro and micronutrient metal elements from animal feeds prior to flame atomic absorption spectrometry. AB - A method for routine analysis of essential metal elements in animal feeds using a dynamic and automated ultrasound-assisted extractor is proposed here. Owing to the different concentration of the essential metal elements in the samples, two macronutrients-namely, Ca and Mg-and three micronutrients or trace elements namely, Fe, Cu and Zn-were selected as models for the optimisation and characterisation of the proposed method by using experimental design methodology. Moreover, the extraction kinetics and influence of the particle size on the extraction efficiency were studied. The detection and quantification limits ranged between 2.9 and 40 microg kg(-1) and between 7.9 and 95.5 microg kg(-1), respectively. The precision, expressed as repeatability relative standard deviation (RSD) and as within-laboratory RSD, ranged between 1.86 and 5.66% and between 5.06 and 6.15%, respectively. The proposed approach allows the extraction of these metal elements from animal feeds with extraction efficiencies similar to those provided by the AOAC Official Method 968.08, but with a drastic reduction in both the extraction time (18 min versus 4.5 h) and sample handling, and using smaller volumes of extractant (an acid aqueous solution). PMID- 14745475 TI - Two new marine sediment standard reference materials (SRMs) for the determination of organic contaminants. AB - Two new marine sediment standard reference materials (SRMs), SRM 1941b Organics in Marine Sediment and SRM 1944 New York/New Jersey Waterway Sediment, have been recently issued by the National Institute of Standards and Technology (NIST) for the determination of organic contaminants including polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyl (PCB) congeners, and chlorinated pesticides. Both sediment SRMs were analyzed using multiple analytical methods including gas chromatography/mass spectrometry (GC/MS) on columns with different selectivity, reversed-phase liquid chromatography with fluorescence detection (for PAHs only), and GC with electron capture detection (for PCBs and pesticides only). SRM 1941b has certified concentrations for 24 PAHs, 29 PCB congeners, and 7 pesticides, and SRM 1944 has certified concentrations for 24 PAHs, 29 PCB congeners, and 4 pesticides. Reference concentrations are also provided for an additional 58 (SRM 1941b) and 39 (SRM 1944) PAHs, PCB congeners, and pesticides. SRM 1944, which was collected from multiple sites within New York/New Jersey coastal waterways, has contaminant concentrations that are generally a factor of 10-20 greater than SRM 1941b, which was collected in the Baltimore (Maryland) harbor. These two SRMs represent the most extensively characterized marine sediment certified reference materials available for the determination of organic contaminants. PMID- 14745476 TI - Metrological assessment of the high-accuracy RNAA method for determination of cobalt in biological materials. AB - The paper summarizes work on the development of the high-accuracy RNAA method for the determination of trace amounts of cobalt in biological materials. The method is based on a combination of neutron activation with selective and quantitative isolation of the analyte in a state of high radiochemical purity by use of column chromatography followed by gamma-ray spectrometric measurements. The method was devised according to a set of rules, which were formulated to obtain high accuracy of the method. The procedure has been also equipped with several criteria as key factors in quality assurance. Qualification of the high-accuracy RNAA method as a primary ratio method has been demonstrated and its usefulness in the certification of the candidate reference materials tea leaves and mixed Polish herbs is presented. PMID- 14745483 TI - Fos immunocytochemical studies on the neuroanatomical sites of action of acute tyrosine depletion in the rat brain. AB - RATIONALE: Acute depletion of brain tyrosine using a tyrosine-free amino acid mixture offers a nutritional approach to reduce central catecholamine function. Recent preclinical data suggest that tyrosine-free amino acid mixtures may have region-specific effects through targeting dopamine neurones. OBJECTIVES: Here we used fos immunocytochemistry to examine the neuroanatomical sites of action of a tyrosine-free amino acid mixture administered either alone or combined with amphetamine. METHODS: Rats (male, Sprague Dawley, 240-260 g) were administered (IP) either a tyrosine-free amino acid mixture (1 g/kg), or the same mixture supplemented with tyrosine and phenylalanine (1 g/kg). Mixtures were injected twice (1 h apart) followed 1 h later by amphetamine (2 mg/kg SC). Two hours later, cardiac perfusion was performed and brains were processed for fos immunocytochemistry. Fos positive cells were counted using computer imaging software. RESULTS: The tyrosine-free amino acid mixture alone did not alter fos expression in ten regions of the rat forebrain compared to saline controls. However, the mixture reduced the increase in fos expression evoked by amphetamine. This effect was region-specific and was greatest in caudate putamen, nucleus accumbens, bed nucleus stria terminalis and lateral habenula, and lacking in other areas including cingulate and insular cortices, lateral septum and central amygdaloid nucleus. Moreover, in most regions the effect of the tyrosine free mixture was less after tyrosine and phenylalanine supplementation. CONCLUSIONS: In summary, a tyrosine-free amino acid mixture reduced amphetamine induced fos expression but this effect was region-specific and included dopamine rich regions. These data further support the idea that tyrosine depletion strategies have potential as treatments for mania and other hyperdopaminergic states. PMID- 14745484 TI - Protein phosphorylation on tyrosine in bacteria. AB - Protein phosphorylation on tyrosine has been demonstrated to occur in a wide array of bacterial species and appears to be ubiquitous among prokaryotes. This covalent modification is catalyzed by autophosphorylating ATP-dependent protein tyrosine kinases that exhibit structural and functional features similar, but not identical, to those of their eukaryotic counterparts. The reversibility of the reaction is effected by two main classes of protein-tyrosine phosphatases: one includes conventional eukaryotic-like phosphatases and dual-specific phosphatases, and the other comprises acidic phosphatases of low molecular weight. Less frequently, a third class concerns enzymes of the polymerase histidinol phosphatase type. In terms of genomic organization, the genes encoding a protein-tyrosine phosphatase and a protein-tyrosine kinase in a bacterial species are most often located next to each other on the chromosome. In addition, these genes are generally part of large operons that direct the coordinate synthesis of proteins involved in the production or regulation of exopolysaccharides and capsular polysaccharides. Recent data provide evidence that there exists a direct relationship between the reversible phosphorylation of proteins on tyrosine and the production of these polysaccharidic polymers, which are also known to be important virulence factors. Therefore, a new concept has emerged suggesting the existence of a biological link between protein-tyrosine phosphorylation and bacterial pathogenicity. PMID- 14745485 TI - Thermobaculum terrenum gen. nov., sp. nov.: a non-phototrophic gram-positive thermophile representing an environmental clone group related to the Chloroflexi (green non-sulfur bacteria) and Thermomicrobia. AB - A novel bacterium was cultivated from an extreme thermal soil in Yellowstone National Park, Wyoming, USA, that at the time of sampling had a pH of 3.9 and a temperature range of 65-92 degrees C. This organism was found to be an obligate aerobic, non-spore-forming rod, and formed pink-colored colonies. Phylogenetic analysis of the 16S rRNA gene sequence placed this organism in a clade composed entirely of environmental clones most closely related to the phyla Chloroflexi and Thermomicrobia. This bacterium stained gram-positive, contained a novel fatty acid profile, had cell wall muramic acid content similar to that of Bacillus subtilis (significantly greater than Escherichia coli), and failed to display a lipopolysaccharide profile in SDS-polyacrylamide gels that would be indicative of a gram-negative cell wall structure. Ultrastructure examinations with transmission electron microscopy showed a thick cell wall (approximately 34 nm wide) external to a cytoplasmic membrane. The organism was not motile under the culture conditions used, and electron microscopic examination showed no evidence of flagella. Genomic G+C content was 56.4 mol%, and growth was optimal at 67 degrees C and at a pH of 7.0. This organism was able to grow heterotrophically on various carbon compounds, would use only oxygen as an electron acceptor, and its growth was not affected by light. A new species of a novel genus is proposed, with YNP1(T) (T=type strain) being Thermobaculum terrenum gen. nov., sp. nov. (16S rDNA gene GenBank accession AF391972). This bacterium has been deposited in the American Type Culture Collection (ATCC BAA-798) and the University of Oregon Culture Collection of Microorganisms from Extreme Environments (CCMEE 7001). PMID- 14745486 TI - Broadband ultrasound attenuation (BUA) of the heel bone and its correlates in men and women in the EPIC-Norfolk cohort: a cross-sectional population-based study. AB - Osteoporotic fractures have substantial clinical and public health impact. Bone quality is an important determinant of fracture risk. Quantitative ultrasound (QUS) of bone measured as broadband ultrasound attenuation (BUA) has been shown to predict fracture risk. However, there have been very few large population studies, particularly in men. We investigated the correlates of calcaneal BUA using a CUBA clinical machine in 15,668 middle and older aged men and women (42 82 years) from the UK, EPIC-Norfolk cohort. At all ages mean BUA was significantly greater in men than women (men, 90.1+/-17.6; women 72.1+/-16.5). The age-related decline in BUA was five times greater in women than men (-0.77 vs. -0.15 dB/MHz per year). Pre- and post-menopausal bone loss was 0.39 and 0.85 dB/MHz per year, respectively. In univariate regression BUA increased with weight and height by 0.45 dB/MHz per kg and 0.68 per cm in women and 0.24 dB/MHz per kg and 0.33 per cm in men. BUA increased with body mass index (BMI) by 0.84 dB/MHz per kg/m2 in women and 0.55 in men. However, weight was twice as influential as height in men and seven times as great in women. Age, weight and height explained 27% of the variance of BUA in women, but only 3% in men. Adjusted BUA was significantly lower in men and women with an existing history of any hip, wrist or spinal fracture both overall and when analysed for specific site. Figures were: all fractures 66.8 vs. 72.5 dB/MHz ( P<0.001), women; 84.1 vs. 90.5 ( P<0.001), men; hip fractures 61.9 vs. 72.2 dB/MHz ( P<0.001), women; 81.5 vs. 90.2 ( P<0.001), men; wrist fractures 66.6 vs. 72.5 dB/MHz ( P<0.001), women; 81.5 vs. 90.2 ( P<0.001), men; spinal fractures 68.1 vs. 72.1 dB/MHz ( P<0.01), women; 85.1 vs. 90.2 ( P<0.01), men. These differences equate to reductions of 14, 9 and 6% and 10, 7 and 6% for fractures of the hip, wrist and spine in the BUA of women and men, respectively. Thus, despite the overall gender difference in BUA the relative magnitude of a previous history of fracture was equally important in both men and women. Adjusted BUA was also lower in those with previous history of osteoporosis. In women currently taking hormone replacement therapy (HRT) the adjusted BUA was 5 dB/MHz or one-third of an SD greater than in those who did not. The BUA of those with a current smoking habit was 1.7% lower in women and 3.2% lower in men. Overall, there are substantial sex differences in the relationship of the physical and osteoporotic risk factors associated with BUA. A better understanding of these determinants of heel ultrasound may provide insights into how some of the sex differences in bone health can be explained and bone loss in later life minimised. PMID- 14745487 TI - Management of osteoporosis in general practice: a cross-sectional survey of primary care practitioners in Spain. AB - A cross-sectional survey was conducted to determine the current situation in Spain regarding diagnosis and care of patients with osteoporosis in the primary care setting. A total of 2,500 primary care physicians who were homogeneously grouped in autonomous communities throughout the country received a postal 30 item anonymous self-administered questionnaire. The questionnaire covered demographics and personal characteristics of the physicians, conditions in everyday consultation, and degree of knowledge with regard to risk factors, diagnosis, treatment, and follow-up of the disease. The overall response was 850 (34%). The mean age of physicians surveyed was 43 years (range 23-66 years). The percentage of physicians specialized in community and family medicine was 46.7%. In 55.2% of cases, years of practice ranged between 11 and 20, and 55.7% of physicians visited between 31 and 50 patients per day. Age and years of practice were not associated with daily number of visits. Only 4% of physicians stated that there were specific programs for osteoporosis implemented in their primary care center. Diagnostic complementary investigations that could be ordered included plain radiographs in 96.2% of cases and bone densitometry in 27.8%. Laboratory tests included serum hormones in 61.6% of cases, PTH in 50.2%, and bone alkaline phosphatase in 33.4%. The diagnosis of osteoporosis was made always personally in 25.2% of cases. Personal diagnosis and follow-up, as well as actions directed to detection of osteoporosis were significantly higher among physicians working in centers with specific programs for osteoporosis. With regard to knowledge about osteoporosis, the mean percentage of correct responses was 63%. The percentage of correct responses was inversely associated with age and years of practice, and positively associated with speciality of community and family medicine. Primary care providers are in a good position to assess risk factors and recommend prevention strategies, as well as to play an active role in the diagnosis, care, and follow-up of patients with osteoporosis. Practitioners of younger age and relatively few years of practice were those with more up-to date information regarding the disease, and the existence of a specific program for osteoporosis seems to improve the management of this condition. PMID- 14745488 TI - Re-use of a liver graft and multi-organ procurement from a liver transplant patient. PMID- 14745489 TI - Aminopyrine breath test compared to the MELD and Child-Pugh scores for predicting mortality among cirrhotic patients awaiting liver transplantation. AB - Better tools for predicting the risk of death while awaiting transplantation are urgently needed because organ shortage is increasing the numbers on transplantation waiting lists. The aminopyrine breath test (ABT), model for end stage liver disease (MELD), and Child-Pugh (C-P) score were compared as predictors of this risk in 137 cirrhotic candidates for liver transplantation. Eighty-three were transplanted within 3 months of registration, 35 others survived, 13 died before transplantation, and 6 were removed from the list. By univariate analysis, the continuous variables significantly associated with death while awaiting transplantation were: history of infected ascites, C-P score, ABT, and international normalized ratio or prothrombin time. Receiver operating characteristic curves for quantitative variables showed that the area under the curve was greatest for ABT (0.858 +/- 0.067). By Youden curve analysis, the best cut-off points for identifying cirrhotic patients at high risk of death while on the waiting list were: > 10, > 16, and < 0.7% for the C-P score, MELD score, and ABT, respectively. These results show that ABT is as good as the MELD and C-P scores, or better, as a predictor of death among cirrhotic patients awaiting liver transplantation. PMID- 14745490 TI - A survey of the public attitudes towards organ donation in a Turkish community and of the changes that have taken place in the last 12 years. AB - In 1990 we carried out a survey on public attitudes toward organ donation in a Turkish community. We repeated this study 12 years later in order to evaluate the changes that had taken place in the meantime. Using the same questionnaire and method, we repeated the study in a different part of the city with similar socio economic characteristics as in the former area, which had in the meantime ceased to be our research and training area. The 983 participants were chosen by a random stratified method. Of those interviewed, 57.0% were willing to donate, while 18.3% refused and 24.7% were uncertain. A total of 52.6% consented to donation. Twelve years later, some public attitudes toward organ donation had changed. Refusal to donate for religious reasons had diminished (16.1% versus 26.2%); uncertainty whether to donate had risen (24.7% versus 15.8%). Attitudes towards organ donation were clearly related to educational level, age and sex. PMID- 14745492 TI - The Golgi apparatus--still causing problems after all these years! PMID- 14745493 TI - ER-to-Golgi transport and cytoskeletal interactions in animal cells. AB - The endoplasmic reticulum (ER)-Golgi system has been studied using biochemical, genetic, electron and light microscopic techniques. We now understand many aspects of trafficking from the ER to the Golgi apparatus, including some of the signals and mechanisms for selective retention and retrieval of ER resident proteins and export of cargo proteins. Proteins that leave the ER emerge in 'export complexes' or ER 'exit sites' and accumulate in pleiomorphic transport carriers referred to sometimes as VTCs or intermediate compartments. These structures then transit from the ER to the Golgi apparatus along microtubules using the dynein/dynactin motor and fuse with the cis cisterna of the Golgi apparatus. Many proteins (including vSNAREs, ERGIC53/p58 and the KDEL receptor) must cycle back to the ER from pre-Golgi intermediates or the Golgi. We will discuss both the currently favored model that this cycling occurs via 50-nm COPI coated vesicles and in vivo evidence that suggests retrograde trafficking may occur via tubular structures. PMID- 14745491 TI - Pro- and anti-inflammatory cytokine production by autoimmune T cells against preproinsulin in HLA-DRB1*04, DQ8 Type 1 diabetes. AB - AIMS/HYPOTHESIS: Preproinsulin is a target T cell autoantigen in human Type 1 diabetes. This study analyses the phenotype and epitope recognition of preproinsulin reactive T cells in subjects with a high genetic risk of diabetes [HLA-DRB1*04, DQ8 with Ab+ (autoantibody-positive) or without islet autoantibodies (control subjects)], and in HLA-matched diabetic patients. METHODS: A preproinsulin peptide library approach was used to screen for cytokine profiles and epitope specificities in human peripheral blood lymphocytes, and CD4(+)CD45RA(-) and CD4(+)CD45RA(+) T cell subfractions, representing memory and naive and recently primed T cells respectively. RESULTS: In CD4(+) T cell subsets we identified immunodominant epitopes and cytokine production patterns that differed profoundly between patients, Ab+ subjects and non-diabetic HLA-matched control subjects. In Ab+ subjects, a C-peptide epitope C13-29 and insulin B-chain epitope B11-27 were preferentially recognised, whereas insulin-treated Type 1 diabetic patients reacted to native insulin and B-chain epitope B1-16. In peripheral blood lymphocytes of Ab+ subjects, an increase in T helper (Th) 1 (IFNgamma, IL-2) and Th2 (IL-4) cytokines was detectable, wheras in CD45RA(+) and CD45RA(-) subsets, IL-4 and IL-10 phenotypes dominated, compatible with the contribution of non-CD4 cells to IFNgamma content. In insulin-treated Type 1 diabetic patients, naive and recently primed CD4(+) cells were characterised by increasd IFNgamma, TNFalpha, and IL-5. CONCLUSIONS/INTERPRETATION: Our data show that T cell reactivity to preproinsulin in CD45RA subsets is Th2-dominant in Ab+ subjects, challenging the Th1 paradigm in Type 1 diabetes. Characteristic immunodominant epitopes and cytokine patterns distinguish diabetic patients and Ab+ subjects from HLA-matched healthy individuals. This could prove useful in monitoring of T-cell immunity in clinical diabetes intervention trials. PMID- 14745494 TI - Structural aspects of Golgi function. AB - Since its discovery, the Golgi complex has attracted the attention of cell biologists because of its 'fashionable' morphology and central position within the secretory system of the cell. Here, we discuss how the three-dimensional architecture of the Golgi complex relates to its multiple functions in protein sorting and processing, and how an analysis of the morphology of the Golgi complex can help to provide an understanding of the mechanisms involved in transport through this unique organelle. PMID- 14745495 TI - Protein localization in the plant Golgi apparatus and the trans-Golgi network. AB - This review presents plant-specific characteristics of the Golgi apparatus and discusses their impact on retention of membrane proteins in the Golgi or the trans-Golgi network (TGN). The plant Golgi consists of distinct stacks of cisternae that actively move throughout the cytoplasm. The Golgi apparatus is a very dynamic compartment and the site for maturation of N-linked glycans. It is also a factory for complex carbohydrates that are part of the cell wall. The TGN is believed to be the site from where vacuolar proteins are sorted by receptors towards each type of vacuole. To maintain the structure and specific features of the Golgi, resident proteins ought to be maintained in the proper Golgi cisternae or in the TGN. Two families of membrane proteins will be taken as examples for Golgi/TGN retention: (i) the enzymes involved in N-glycosylation processes and (ii) a vacuolar sorting receptor. Although the number of available plant proteins localized in Golgi/TGN is low, the basis of retention appears to be shared over all kingdoms and may result from pure retention and recycling mechanisms. In this review, we will summarize the characteristics of a plant Golgi and will discuss especially their consequences on on the study of this highly dynamic structure. We then choose membrane proteins with a single transmembrane domain to illustrate the signals and mechanisms involved in plants to localize and maintain proteins in the Golgi and the TGN. PMID- 14745496 TI - Dynamics of proteins in Golgi membranes: comparisons between mammalian and plant cells highlighted by photobleaching techniques. AB - In less than a decade the green fluorescent protein (GFP) has become one of the most popular tools for cell biologists for the study of dynamic processes in vivo. GFP has revolutionised the scientific approach for the study of vital organelles, such as the Golgi apparatus. As Golgi proteins can be tagged with GFP, in most cases without altering their targeting and function, it is a great substitute to conventional dyes used in the past to highlight this compartment. In this review, we cover the application of GFP and its spectral derivatives in the study of Golgi dynamics in mammalian and plant cells. In particular, we focus on the technique of selective photobleaching known as fluorescence recovery after photobleaching, which has successfully shed light on essential differences in the biology of the Golgi apparatus in mammalian and plant cells. PMID- 14745497 TI - Yeast Golgi apparatus--dynamics and sorting. AB - The yeast Saccharomyces cerevisiae is one of the best-studied organisms to understand molecular mechanisms of membrane traffic, but as far as the organization of the Golgi apparatus is concerned, yeast is only just beginning to yield clues about how dynamic and flexible the organelle is. PMID- 14745498 TI - Diversity of structures and properties among catalases. AB - More than 300 catalase sequences are now available, divided among monofunctional catalases (> 225), bifunctional catalase-peroxidases (> 50) and manganese containing catalases (> 25). When combined with the recent appearance of crystal structures from at least two representatives from each of these groups (nine from the monofunctional catalases), valuable insights into the catalatic reaction mechanism in its various forms and into catalase evolution have been gained. The structures have revealed an unusually large number of modifications unique to catalases, a result of interacting with reactive oxygen species. Biochemical and physiological characterization of catalases from many different organisms has revealed a surprisingly wide range of catalatic efficiencies, despite similar sequences. Catalase gene expression in micro-organisms generally is controlled either by sensors of reactive oxygen species or by growth phase regulons, although the detailed mechanisms vary considerably. PMID- 14745499 TI - Hopes, disillusions and more hopes from vitamin C. AB - In the current view of most biochemists and physiologists, the role of L-ascorbic acid (AA) in cell metabolism would be more or less confined to the scavenging of reactive oxygen species. Nevertheless, many data have been collected in our and other laboratories concerning the involvement of AA in many different aspects of cell metabolism. At the present time the molecular sites of action of AA have not been completely elucidated, but recent findings on the specific requirement of AA for the activity of several 2-oxoacid-dependent dioxygenases involved in cell signalling and the activation of transcription factors open new fascinating perspectives for further research. PMID- 14745500 TI - Mitochondrial respiratory function and antioxidant capacity in normal and cirrhotic livers following partial hepatectomy. AB - For many liver malignancies, major hepatectomy is the usual therapy. Although a normal liver has a tremendous capacity for regeneration, liver hepatectomy in humans is usually carried out on a diseased liver and, in such cases, liver regeneration takes place in a cirrhotic remnant. Mitochondrial function in cirrhotic livers shows a variety of changes compared to control livers. This study investigated how mitochondrial respiratory function and antioxidant capacity change following partial hepatectomy of cirrhotic livers, because liver regeneration requires greater energy demands and control of oxidative stress. Cirrhosis was induced in male Wistar-Furth rats by administration of thioacetamide. NADH-cytochrome c reductase activity, mitochondrial glutathione peroxidase activity and mitochondrial GSH levels were all significantly lowered in cirrhotic livers and in the cirrhotic remnants up to 72 h after 70% hepatectomy when compared to the corresponding controls. Lower respiratory control ratios with succinate as substrate were also observed from 6 to 48 h post hepatectomy. At 24 h post-hepatectomy, higher levels of lipid peroxidation were observed. We conclude that, compared to the controls, cirrhotic livers have diminished oxidative phosphorylation capabilities due to changes in NADH and FADH(2)-linked respiration as well as impaired antioxidant defenses following partial hepatectomy. Both of these factors, if critical, could then impede liver regeneration. PMID- 14745501 TI - Nucleotide-binding domains of human cystic fibrosis transmembrane conductance regulator: detailed sequence analysis and three-dimensional modeling of the heterodimer. AB - The cystic fibrosis transmembrane conductance regulator (CFTR) protein is encoded by the gene that is defective in cystic fibrosis, the most common lethal inherited disease among the Caucasian population. CFTR belongs to the ABC transporter superfamily, whose members form macromolecular architectures composed of two membrane-spanning domains and two nucleotide-binding domains (NBDs). The experimental structures of NBDs from several ABC transporters have recently been solved, opening new avenues for understanding the structure/function relationships and the consequences of some disease-causing mutations of CFTR. Based on a detailed sequence/structure analysis, we propose here a three dimensional model of the human CFTR NBD heterodimer. This model, which is in agreement with recent experimental data, highlights the specific features of the CFTR asymmetric active sites located at the interface between the two NBDs. Moreover, additional CFTR-specific features can be identified at the subunit interface, which may play critical roles in active site interdependence and are uncommon in other NBD dimers. PMID- 14745502 TI - Aspartate 338 contributes to the cationic specificity and to driver-amino acid coupling in the insect cotransporter KAAT1. AB - To investigate the peculiar ionic specificity of KAAT1, an Na+- and K+-coupled amino acid cotransporter from Lepidoptera, a detailed analysis of membrane topology predictions was performed, together with sequence comparison with strictly Na+-dependent mammalian cotransporters from the same family. The analysis identified aspartate 338, a residue present also in the other cotransporter accepting K+ (CAATCH1), but absent in most mammalian transporters that have, instead, an asparagine in the corresponding position. Mutation of D338 in KAAT1 led either to non-functional transporters (D338G, D338C), or to an altered ionic selectivity (D338E, D338N), observable in uptake experiments and in electrophysiological properties. In particular, in D338E, the transport activity, while persisting in the presence of Na+, appeared to be completely abolished in the presence of K+. D338E also showed uncoupling between transport-associated current and uptake. The opposite mutation in the gamma-aminobutyric acid transporter rGAT-1 (N327D) resulted in complete loss of function. In conclusion, aspartate 338 in KAAT1 appears to be important in allowing K+, in addition to Na+, to drive the transport mechanism, although other residues in different parts of the protein may also play a role in the complete determination of ionic selectivity. PMID- 14745503 TI - Agrobacterium rhizogenes-mediated DNA transfer to Aesculus hippocastanum L. and the regeneration of transformed plants. AB - Hairy roots were induced from androgenic embryos of horse chestnut (Aesculus hippocastanum L.) by infection with Agrobacterium rhizogenes strain A4GUS. Single roots were selected according to their morphology in the absence of antibiotic or herbicide resistance markers. Seventy-one putative transformed hairy root lines from independent transformation events were established. Regeneration was induced in MS liquid medium supplemented with 30 microM 6-benzylaminopurine (BA), and the regenerants were multiplied on MS solid medium containing 10 microM BA. Following elongation on MS medium supplemented with 1 microM BA and 500 mg/l polyvinylpyrrolidone, the shoots were subjected to a root-inducing treatment. Stable integration of TL-DNA within the horse chestnut genome was confirmed by Southern hybridization. The copy number of transgenes was estimated to be from two to four. PMID- 14745504 TI - Interspecific hybridization of Trifolium alexandrinum with T. constantinopolitanum using embryo rescue. AB - The embryo rescue technique was successfully used to raise hybrids between Trifolium alexandrinum and T. constantinopolitanum. As a result of its narrow genetic base, genetic improvement in Egyptian clover (syn. Berseem; T. alexandrinum), an important fodder crop in tropical and subtropical countries, is hampered, thereby making it imperative to introduce alien genes from related species. In a conventional interspecific hybridization program, hybrids could not be raised due to post-fertilization barriers. Of the several combinations tried, pollination 2 days after emasculation was found to be the best. Globular embryos were observed 5-7 days after pollination (DAP), followed by heart-shaped embryos 10-12 DAP. Embryos excised at the heart-shaped stage responded well to EC3 culture medium. Of 612 crosses, 33 healthy embryos could be excised and cultured on EC3 medium. The plumule emerged 8-12 days following inoculation. The embryo rescued plants were hardened, inoculated with Rhizobium and transferred to the field. The hybrids showed intermediate morphological features with reduced pollen fertility (55-65%) and a chromosomal complement of 2n = 16. Biochemical characterization using isozymes confirmed hybridity. PMID- 14745505 TI - The prevalence of joint hypermobility among high school students. AB - OBJECTIVE: The aim of this study was to determine the prevalence of joint hypermobility among high school students and to define the characteristics of patients with joint hypermobility. METHODS: The students underwent complete history and physical examination. In order to designate marfanoid habitus, body weight, height, and span/height and upper/lower segment ratios were recorded. The degree of joint hypermobility was scored by the Beighton scoring system. The following features were also examined: arthralgia, myalgia, low back pain, sciatica, spinal deformities, temporomandibular joint pain and crepitus, effusion, swan neck deformity, arachnodactyly, joint dislocation, joint sprain, Raynaud's phenomenon, stria, varicose veins, abdominal and inguinal hernia, heart disease history, myopia, dropping eyelids, and antimongoloid slant. RESULTS: Eight hundred sixty-one students (433 females and 428 males) with a mean age of 15.4+/-1.1 years (range 13-19) were examined. Joint hypermobility was observed in 101 (11.7%) of the students. According to the Beighton scoring system, the majority of these (61.4%) were observed to score 4. Our results show that phenotype has no relation with joint mobility. Of the total number of students, there were 31 male (7.2%) and 70 female (16.2%) hypermobile subjects. The difference between sexes was highly significant (P=0.00005). Joint sprain was detected in 14 of hypermobile students (13.9%) and 50 of nonhypermobile students (6.6%). Its presence was the only significant parameter between hypermobile and nonhypermobile students (P=0.0094). CONCLUSIONS: Joint hypermobility was found in 11.7% of the students in our study, and the results are in harmony with the previous studies on Western populations. Although hypermobility does not seem to be very problematic in young people, as in our focus group, we believe that it is important for physicians to recognize this problem to ensure correct diagnosis and treatment, since it may lead to mimic rheumatic diseases in the future. PMID- 14745506 TI - A one-step method to convert vectors into binary vectors suited for Agrobacterium mediated transformation. AB - Bacterial artificial chromosomes (BACs) are widely used for the construction of physical maps, positional-cloning and whole-genome sequencing strategies. Unfortunately, their use for functional genomics is limited, as currently there is no efficient method to use BACs directly for complementation. We describe a novel strategy for one-step conversion of any BAC into a binary BAC (BIBAC). Using Agrobacterium tumefaciens, these BIBACs can be efficiently transformed to virtually all organisms, including plants, fungi, yeasts and human cells. As the strategy is based on in vivo recombineering and does not depend on restriction sites, it is applicable to any vector. To show the feasibility of the method five BACs, containing 0-75 kb of fungal DNA, were converted into BIBACs. These were subsequently transformed to the plant pathogenic fungus Fusarium oxysporum f.sp. lycopersici and to Aspergillus awamori, a filamentous fungus often used for large scale protein production. Molecular characterisation of the transformants showed that the BIBACs were efficiently transferred to the fungi and stably integrated into their genomes. PMID- 14745507 TI - The role of non-viraemic transmission on the persistence and dynamics of a tick borne virus--Louping ill in red grouse ( Lagopus lagopus scoticus) and mountain hares ( Lepus timidus). AB - There exist many tick borne infections that are of either economic or public health interest. Mathematical models have previously been used to describe the dynamics of these infections. However it has recently come to light that there is an alternative mechanism for the transmission of these diseases that has not been considered in a modelling framework. This is transmission through ticks co feeding on non-viraemic hosts. This paper extends a simple mathematical model to include this alternative transmission mechanism. The model is used to describe the dynamics of Louping ill virus in red grouse (the viraemic host) and hares (the non-viraemic host). However, these results are applicable to many other systems. The model is analysed using joint threshold density curves. It is found that the presence of a non-viraemic host allows the virus to persist more readily than it would in the presence of a host that simply amplified the tick population. More importantly, if the level of non-viraemic transmission is high enough the virus can persist in the absence of the viraemic host. This result has important implications for the control of tick borne diseases. PMID- 14745508 TI - Small mutation rate and evolutionarily stable strategies in infinite dimensional adaptive dynamics. AB - An integrodifferential equations model for the distribution of individuals with respect to the age at maturity is considered. Mutation is modeled by an integral operator. Results concerning the behaviour of the steady states and their relation to evolutionarily stable strategies when the mutation rate is small are given. The same results are obtained for a (rather) general class of models that include the one mentioned before. PMID- 14745509 TI - On the minimum number of recombination events in the evolutionary history of DNA sequences. AB - In representing the evolutionary history of a set of binary DNA sequences by a connected graph, a set theoretical approach is introduced for studying recombination events. We show that set theoretical constraints have direct implications on the number of recombination events. We define a new lower bound on the number of recombination events and demonstrate the usefulness of our new approach through several explicit examples. PMID- 14745510 TI - Deriving reaction-diffusion models in ecology from interacting particle systems. AB - We use a scaling procedure based on averaging Poisson distributed random variables to derive population level models from local models of interactions between individuals. The procedure is suggested by using the idea of hydrodynamic limits to derive reaction-diffusion models for population interactions from interacting particle systems. The scaling procedure is formal in the sense that we do not address the issue of proving that it converges; instead we focus on methods for computing the results of the scaling or deriving properties of rescaled systems. To that end we treat the scaling procedure as a transform, in analogy with the Laplace or Fourier transform, and derive operational formulas to aid in the computation of rescaled systems or the derivation of their properties. Since the limiting procedure is adapted from work by Durrett and Levin, we refer to the transform as the Durrett-Levin transform. We examine the effects of rescaling in various standard models, including Lotka-Volterra models, Holling type predator-prey models, and ratio-dependent models. The effects of scaling are mostly quantitative in models with smooth interaction terms, but ratio-dependent models are profoundly affected by the scaling. The scaling transforms ratio dependent terms that are singular at the origin into smooth terms. Removing the singularity at the origin eliminates some of the unique dynamics that can arise in ratio-dependent models. PMID- 14745511 TI - Macrophage response to Mycobacterium tuberculosis infection. AB - The immune response to Mycobacterium tuberculosis (Mtb) infection is the formation of multicellular lesions, or granolomas, in the lung of the individual. However, the structure of the granulomas and the spatial distribution of the immune cells within is not well understood. In this paper we develop a mathematical model investigating the early and initial immune response to Mtb. The model consists of coupled reaction-diffusion-advection partial differential equations governing the dynamics of the relevant macrophage and bacteria populations and a bacteria-produced chemokine. Our novel application of mathematical concepts of internal states and internal velocity allows us to begin to study this unique immunological structure. Volume changes resulting from proliferation and death terms generate a velocity field by which all cells are transported within the forming granuloma. We present numerical results for two distinct infection outcomes: controlled and uncontrolled granuloma growth. Using a simplified model we are able to analytically determine conditions under which the bacteria population decreases, representing early clearance of infection, or grows, representing the initial stages of granuloma formation. PMID- 14745515 TI - Identification of an antigenic peptide derived from the cancer-testis antigen NY ESO-1 binding to a broad range of HLA-DR subtypes. AB - NY-ESO-1 is a SEREX-defined cancer-testis antigen of which several MHC I, but only few MHC II-restricted epitopes have been identified. Searching for highly promiscuous MHC II-restricted peptides that might be suitable as a CD4+ stimulating vaccine for many patients, we used the SYFPEITHI algorithm and identified an NY-ESO-1-derived pentadecamer epitope (p134-148) that induced specific CD4+ T-cell responses restricted to the HLA-DRB1 subtypes *0101, *0301, *0401, and *0701 that have a cumulative prevalence of 40% in the Caucasian population. The DR restriction of the p134-148 pentadecamer was demonstrated by inhibition with an HLA-DR antibody and a functional peptide displacement titration assay with the pan-DR-binding T-helper epitope PADRE as the competitor. The natural processing and presentation of this epitope was demonstrated by recognition of an NY-ESO-1+ melanoma cell line by T cells with specificity for p134-148. The pentadecamer p134-148 was able to induce CD4+ responses in 4/38 cancer patients tested. However, no strict correlation was found between CD4+ T cell responses against p134-148 reactivity and anti-NY-ESO-1 antibody titers in the serum of patients, suggesting that CD4+ and B-cell responses are regulated independently. In conclusion, p134-148 holds promise as a broadly applicable peptide vaccine for patients with NY-ESO-1-positive neoplasms. PMID- 14745516 TI - Targeting of lacZ reporter gene expression with radioiodine-labelled phenylethyl beta- d-thiogalactopyranoside. AB - There has recently been increasing interest in the development of radioprobes that specifically target proteins transcribed from expression of reporter genes of interest. The purpose of this study was to develop a radioprobe that targets one of the most widely used reporter genes, the bacterial lacZ gene. We synthesised and purified radioiodine-labelled phenylethyl-beta- d thiogalactopyranoside (PETG), a competitive inhibitor specific against Escherichia coli beta-galactosidase. We showed that [(125)I]iodo-PETG specifically binds to beta-galactosidase as verified by column chromatography and polyacrylamide gel electrophoresis after incubation of radiotracer with the protein. We also showed through enzyme kinetic studies that iodo-PETG retains inhibitory action against beta-galactosidase activity. COS-7 cells infected with a recombinant adenovirus expressing the lacZ gene had viral titre-dependent enhancements in [(125)I]iodo-PETG uptake ( r(2)=0.897; P=0.001), which reached up to 642.5%+/-16.7% of control levels ( P<0.00001). Moreover, the level of uptake was highly correlated to luminescent measurements of beta-galactosidase activity ( r(2)=0.878; P<0.0001). These results confirm that radioiodine-labelled PETG specifically targets beta-galactosidase and that its uptake rates faithfully reflect levels of expression of the lacZ reporter gene. Further investigations were performed in nude mice bearing human neuroblastoma tumours transferred with the lacZ gene. Compared with control tumours, lacZ-expressing tumours were slightly better visualised on [(123)I]iodo-PETG images and had a modest increase in tumour to muscle count ratio (2.6+/-0.2 vs 1.9+/-0.1, P<0.05). The present results provide proof-of-principle for the potential of radiolabelled inhibitors as promising radiotracers to monitor lacZ gene expression levels. Future modifications to improve cell permeability should enhance in vivo contrast levels and may allow the use of radiolabelled beta-galactosidase inhibitors for non invasive monitoring of lacZ gene expression. PMID- 14745517 TI - Subareolar injection of technetium-99m nanocolloid yields reliable data on the axillary lymph node tumour status in breast cancer patients with previous manipulations on the primary tumour: a prospective study of 117 patients. AB - According to recently published guidelines, histological clarification by interventional techniques should be undertaken before planning the surgical management of patients with breast carcinoma. In patients with previous manipulations on the primary tumour, peritumoural injection in the context of preoperative scintigraphic detection of the sentinel lymph nodes is not possible. The aim of this prospective study was to clarify whether subareolar injection of nanocolloid can yield reliable data on the axillary lymph node tumour status in breast cancer patients with previous manipulations on the primary tumour. To date, 117 women (age 31-80 years) with breast carcinoma have been enrolled. All of these patients had undergone a biopsy (n=88) or surgery on the primary tumour (n=29) and were without clinical suspicion of lymph node metastases. Subareolar injection of 40 MBq technetium-99m nanocolloid was carried out in at least eight deposits around the areolar margin [one deposit in the middle of each quadrant and one deposit at each quadrant intersection (0.05 ml/deposit)]. Immediately after injection, dynamic and static lymphoscintigraphy of the axillary, thoracic and cervical areas was performed in various views with a gamma camera (LEAP collimator, 256x256 matrix). Lymphatic drainage was directed exclusively to the ipsilateral axilla. Sentinel lymph node biopsy and elective dissection of axillary lymph nodes were performed in all patients. All lymph nodes removed were examined by histology and immunohistochemistry. In 26 patients, lymph node metastases were found in the sentinel lymph nodes. In six of them, non-sentinel lymph nodes also showed tumour involvement. In the remaining 91 patients, lymph node metastases could be found neither in sentinel lymph nodes nor in non sentinel lymph nodes. In conclusion, subareolar nanocolloid injection can yield reliable information on the axillary lymph node tumour status in patients with previous manipulations on the primary tumour in the breast. PMID- 14745518 TI - Influence of the substrate on the ultrastructure of Pleurotus pulmonarius fruit body primordia. AB - Pleurotus pulmonarius fruit body primordia (FBP) formed on potato extract agar (PEA) and wheat straw-based media (WS) were studied. When grown on PEA, FBP hyphae had a large number of vacuoles and less stainable cytoplasmic material. In contrast, when grown on WS, there was abundant cytoplasmic material and fewer vacuoles. FBP grown on WS had a more well-defined mushroom shape than those grown on PEA. The average hyphal diameters on PEA and WS were 4.15 and 3.52 micro m, respectively; the average hyphal lengths were 11.75 and 11.80 micro m, respectively. The control mechanisms regulating formation of the normal shape and size of FBP might be more dependent on the amount of cytoplasmic material than on the physical volume of the hyphal compartment. Since the ultrastructure of the hyphal compartment, which makes up the substance of the FBP, depends on the substrate in which the FBP has been grown, the composition of the substrate may be important for manipulating the nutritive and organoleptic characteristics of the mushroom crop. PMID- 14745519 TI - Orally administrable enterotoxigenic Escherichia coli vaccine encapsulated by ethylcellulose powder dispersion. AB - To overcome the limitations of injection administration to vaccinate neonatal piglets against diarrheal disease, an oral vaccine needs to be developed. Enteric microspheres of oral vaccines were developed by a co-spray drying process based on formalin-inactivated enterotoxigenic Escherichia coli antigens with various encapsulating materials. The encapsulating efficiencies of ECN7m, ECN14m and ECN22m (vaccine microsphere formulations) tested by extraction procedure are high, more than 85%. To assess enteric characteristics, an in vitro dissolution test was performed with microspheres. Formulations with ethylcellulose ECN14m and ECN22m allow controlled release in a neutral or basic environment and resisted acid damage. In all cases, 95% of the E. coli protein was released within 2 h at pH 6.8-7, but there was no release at pH 1.5-2. However, ECN7m was less acid resistant and had lower release at low pH. In animal immunization tests, oral immunization with microspheres of formulations ECN14 and ECN22m effectively evoked both systemic IgG and mucosal IgA responses against E. coli whole cell antigens in mice. In the mice challenge test, orally administrable ECNm14 (12 mg) or ECN22m (12.6 mg) vaccine (i.e., encapsulating 3.0x10(9) cfu inactive bacterial mass) provided good protection from infection in animals. PMID- 14745520 TI - Assimilation of grape phytosterols by Saccharomyces cerevisiae and their impact on enological fermentations. AB - Although yeasts are known to be able to incorporate a wide variety of exogenous sterols under strict anaerobiosis, no data are available on the assimilation of grapevine phytosterols under enological conditions and the eventual impact on fermentation kinetics. We used therefore a mixture of pure phytosterols, in a proportion representative of the different grape skins phytosterols, to supplement a synthetic fermentation medium simulating a grape must. Under anaerobiosis, normal biomass formation was achieved with 5 mg phytosterols l(-1). Similar results were obtained in comparison with the observed maximal fermentation rates. These results clearly indicated that grape phytosterols may efficiently act as a substitute for ergosterol in the yeast membrane for promoting yeast growth and initial fermentative activity. Analysis of total yeast sterols indicated that phytosterols are accumulated without further modification, mainly in their esterified form. However, all the fermentations performed with synthetic media supplemented with phytosterols led to stuck fermentations, linked to a correlative strong decrease in cell viability during the stationary phase. Therefore, grape phytosterols are easily incorporated by yeast cells under enological conditions for promoting initial growth and fermentative activity, but rapidly perturb the yeast membrane properties by being the predominant sterols. PMID- 14745521 TI - Rat T-cell receptor TRAV11 (Valpha14) genes: further evidence of extensive multiplicity with homogeneous CDR1 and diversified CDR2 by genomic contig and cDNA analysis. AB - The antigen recognition system of NKT cells acts via an invariant T-cell receptor (TR) which recognizes CD1d and is highly conserved in mice, rats and humans. NKT cells expressing an invariant mouse TR composed of TRAV11-TRAJ18 (formerly Valpha14-Jalpha281) are positively selected by CD1d, and recognize an antigen in context with CD1d. Here we show ten distinct TRAV11 genes (previously designated by us as TRAV14) on rat Chromosome 15 (BN/SsNHsd/MCW strain). In the rat TRAV11 genes, the splicing sites, the recombination signal sequences, and the possible promoter regions were well conserved, indicating that they were functional. Predicted protein sequences of rat TRAV11 genes were analyzed, including the three loops (CDR1-3) which connect the beta-strands of the domain encoded by the TRA V-REGION and is hypervariable in sequence. The CDR1-IMGT sequence (from 27 to 32; VTPFNN) was conserved among most rat TRAV11 genes. The CDR2-IMGT sequences (from 56 to 61) were grouped into two types: type 1 [L(T/K)NKEE], and type 2 [LAYKKE]. The mRNAs of both types have a different tissue distribution. The CDR3 sequences were short and invariant, the rat TRAV11 genes being preferentially rearranged with rat TRAJ18 (Jalpha281), with the joint consisting of a single amino acid (A or G). Thus, rats had multiple TRAV11 chains with diversified CDR2 IMGT and homogenous CDR1-IMGT and CDR3-IMGT. PMID- 14745522 TI - The DY genes of the cattle MHC: expression and comparative analysis of an unusual class II MHC gene pair. AB - The major histocompatibility complex of cattle (BoLA) contains the class II genes DYA and DIB which are transcribed with a dendritic cell restricted distribution. As part of the process to determine whether these genes have any functional significance, we demonstrate that they form a closely linked pair characteristic of other expressed class II MHC molecules. Accepted nomenclature convention suggests that BoLA-DIB should therefore be renamed BoLA-DYB. Analysis of the first full-length DYA and DYB transcripts revealed open reading frames with potential to translate 253 and 259 amino acid proteins, respectively. Comparative sequence analysis between the DY polypeptides and classical cattle, human and mouse class II MHC alpha and beta polypeptide chains revealed 16 unique amino acid residues at positions predicted to form and line the putative peptide binding region. Expression of tagged constructs demonstrates for the first time that the DY genes of cattle are capable of translating distinctive class II MHC alpha and beta polypeptide chains. PMID- 14745524 TI - Thermal and guanidine hydrochloride-induced denaturation of human cystatin C. AB - Wild-type human cystatin C is directly involved in pathological fibrils formation, leading to hemorrhage, dementia and eventually death of people suffering from cerebral amyloid angiopathy. Some studies on cystatin C oligomerization have been already done but some points are still unclear. In order to learn more about this important process, we have investigated thermal and chemical (guanidine hydrochloride-induced) denaturation of human cystatin C. Studies performed using tryptophan fluorescence, calorimetry, circular dichroism and Fourier transform infrared spectroscopy demonstrate that neither chemical nor thermal denaturation of hCC are simple two-state events. One recognized intermediate form was dimeric cystatin C, whose appearance was preceded mainly by changes in the L2 binding loop. The other form occurred only in the chemical denaturation process and was characterized by partially recovered interactions maintaining the protein tertiary structure. Our studies also strongly indicate that the beta-structural motif of cystatin C is directly implicated in formation of temperature-induced aggregates. PMID- 14745523 TI - Is the junctional uncoupling elicited in rat ventricular myocytes by some dephosphorylation treatments due to changes in the phosphorylation status of Cx43? AB - Gap junctions, specialized membrane structures that mediate cell-to-cell communication in almost all animal tissues, are composed of channel-forming integral membrane proteins termed connexins. Most of them, particularly connexin43 (Cx43), the most ubiquitous connexin, the major connexin present in cardiac myocytes, are phosphoproteins. Connexin phosphorylation has been thought to regulate gap junctional protein trafficking, gap junction assembly, channel gating, and turnover. Some connexins, including Cx43, show mobility shifts in gel electrophoresis when cells are exposed to phosphorylating or dephosphorylating treatments. However, after exposure of rat cardiac myocytes to different uncoupling dephosphorylating agents such as H7 or butanedione monoxime, no modification in the Cx43 phosphorylation profile was generally observed. The lack of direct correlation between the inhibition of cell-to-cell communication and changes in the phosphorylation pattern of Cx43 or, conversely, modifications of the latter without modifications of the intercellular coupling degree, suggest that the functional state of junctional channels might rather be determined by regulatory proteins associated with Cx43. The modulation of the activity of junctional channels by protein phosphorylation/dephosphorylation processes very likely requires (as for several other membrane channels) the formation of a multiprotein complex, where pore-forming subunits bind to auxiliary proteins (e.g. scaffolding proteins, enzymes, cytoskeleton elements) that play essential roles in channel localization and activity. Such regulatory proteins, behaving as targets for phosphorylation/dephosphorylation catalysers, might in particular control the open probability of junctional channels. A schematic illustration of the regulation of Cx43-made channels by protein phosphorylation involving a partner phosphoprotein is proposed. PMID- 14745525 TI - Issues specific to implementing PET-CT for pediatric oncology: what we have learned along the way. AB - In parallel with the expansion of PET imaging to pediatric patients has been the technological development of merging state-of-the-art cross-sectional anatomic information (CT) with functional imaging (PET) into a single modality: PET-CT. Attending to the clinical, scheduling, and medical needs that are unique to imaging children and adolescents can be a challenge, particularly when instituting a single new modality. When that modality bridges two unique, previously independent methods-often previously located in two separate departmental divisions-the details and logistics required to set up a smoothly functioning process can be particularly difficult. This paper focuses on our experience in implementing PET-CT in a tertiary pediatric referral center. PMID- 14745526 TI - PET imaging in pediatric oncology. AB - High-quality PET imaging of pediatric patients is challenging and requires attention to issues commonly encountered in the practice of pediatric nuclear medicine, but uncommon to the imaging of adult patients. These include intravenous access, fasting, sedation, consent, and clearance of activity from the urinary tract. This paper discusses some technical differences involved in pediatric PET to enhance the quality of scans and assure the safety and comfort of pediatric patients. PMID- 14745527 TI - Positron emission tomography in pediatric radiation oncology: integration in the treatment-planning process. AB - The application of PET imaging to pediatric radiation oncology allows new approaches to targeting and selection of radiation dose based not only on the size of a tumor, but also on its metabolic activity. In order to integrate PET into treatment planning for radiation oncology, logistical issues regarding patient setup, image fusion, and target selection must be addressed. Through prospective study, the role of PET in pediatric malignancies will be established for diagnosis, treatment, and surveillance. To explore the potential role of PET and its incorporation into treatment planning in pediatric radiation oncology, an example case of pediatric Hodgkin's disease is discussed. PMID- 14745528 TI - PET imaging in pediatric Hodgkin's lymphoma. AB - Advances in diagnostic imaging technology, especially functional imaging modalities like positron emission tomography (PET), have significantly influenced the staging and treatment approaches used for pediatric Hodgkin's lymphoma. Today, the majority of children and adolescents diagnosed with Hodgkin's lymphoma will be cured following treatment with non-cross-resistant combination chemotherapy alone or in combination with low-dose, involved-field radiation. This success produced a greater appreciation of long-term complications related to radiation, chemotherapy, and surgical staging that prompted significant changes in staging and treatment protocols for children and adolescents with Hodgkin's lymphoma. Contemporary treatment for pediatric Hodgkin's lymphoma uses a risk-adapted approach that reduces the number of combination chemotherapy cycles and radiation treatment fields and doses for patients with localized favorable disease presentation. Advances in diagnostic imaging technology have played a critical role in the development of these risk-adapted treatment regimens. The introduction of computed tomography (CT) provided an accurate and non-invasive modality to define nodal involvement below the diaphragm that motivated the change from surgical to clinical staging. The introduction of functional imaging modalities, like positron emission tomography (PET) scanning, provided the means to correlate tumor activity with anatomic features generated by CT and modify treatment based on tumor response. For centers with access to this modality, PET imaging plays an important role in staging, evaluating tumor response, planning radiation treatment fields, and monitoring after completion of therapy for pediatric Hodgkin's lymphoma. This trend will likely increase in the future as a result of PET's superior sensitivity in correlating sites of tumor activity compared to other available functional imaging modalities. Ongoing prospective studies of PET in pediatric patients will increase understanding about the optimal use of this modality in children with cancer and define the characteristics of FDG-avid nonmalignant conditions that may be problematic in the interpretation of tumor activity. PMID- 14745529 TI - Invertebrate species with nonpelagic larvae have elevated levels of nonsynonymous substitutions and reduced nucleotide diversities. AB - Under a nearly neutral model in which most amino acid substitutions are slightly deleterious, variation in demography, population structure, and other ecological factors among closely related species can potentially modify the effective population size or the selective regime, leading to differences in the rate of nonsynonymous substitution. Ratios of nonsynonymous to synonymous substitutions (d(N)/d(S)) between species were analyzed in a sea star genus (Patiriella) and a molluscan genus (Littorina), each with diverse modes of reproduction, including multiple lineages with pelagic and nonpelagic larvae. In both genera, lineages with nonpelagic larvae had significantly higher d(N)/d(S) ratios than lineages with pelagic larvae. The hypothesis that the elevated d(N)/d(S) ratios in species with nonpelagic larvae was due to reduced effective population size was tested by comparing nucleotide diversities in three genera of gastropod mollusks (Littorina, Crepidula, and Hydrobia), each with several modes of reproduction. Overall, there was a significant (p < 0.05) reduction in nucleotide diversity in species with nonpelagic larvae compared to species with pelagic larvae. PMID- 14745530 TI - Comparisons of two large phaeoviral genomes and evolutionary implications. AB - The evolution of viral genomes has recently attracted considerable attention. We compare the sequences of two large viral genomes, EsV-1 and FirrV-1, belonging to the family of phaeoviruses which infect different species of marine brown algae. Although their genomes differ substantially in size, these viruses share similar morphologies and similar latent infection cycles. In fact, sequence comparisons show that the viruses have more than 60% of their genes in common. However, the order of genes is completely different in the two genomes, suggesting that extensive recombinational events in addition to several large deletions had occurred during the separate evolutionary routes from a common ancestor. We investigated genes encoding components of signal transduction pathways and genes encoding replicative functions in more detail. We found that the two genomes possess different, although overlapping, sets of genes in both classes, suggesting that different genes from each class were lost, perhaps randomly, after the separate evolution from an ancestral genome. Random loss would also account for the fact that more than one-third of the genes in one viral genome has no counterparts in the other genome. We speculate that the ancestral genome belonged to a cellular organism that had once invaded a primordial brown algal host. PMID- 14745531 TI - Marked intragenomic heterogeneity and geographical differentiation of nrDNA ITS in Larix potaninii (Pinaceae). AB - Nuclear ribosomal DNA (nrDNA) of gymnosperms, especially Pinaceae, is characterized by slow concerted evolution and exhibits substantial ITS-region length variation (975-3663 bp), in sharp contrast to the narrow range (565-700 bp) in angiosperms. Here we examined intra- and intergenomic heterogeneity of the nrDNA internal transcribed spacer (ITS) region in four varieties of Larix potaninii, a species from the mountainous areas of western China. Two clones with more than a 100-bp deletion in ITS1 were detected in L. potaninii var. chinensis and L. potaninii var. australis, respectively. The deletion resulted in the loss of most part, including the motif sequence, of subrepeat 1 (SR1). Sequence divergence and phylogenetic analyses showed that some clones would be pseudogenes given their low GC content, high substitution rates, unique positions in the phylogenetic trees, or significant length variation. These clones might represent orphons or paralogues at minor loci resulting from large-scale gene or chromosome reorganization. Some recombinants characterized by chimeric structure and discordant phylogenetic positions in partitioned sequence analyses indicate that unequal crossing-over plays an important role in the process of nrDNA concerted evolution. In addition, some varieties of L. potaninii might have experienced an nrDNA founder effect parallel to their geographical isolation. PMID- 14745532 TI - Relative rates of nucleotide substitution in frogs. AB - Accurate estimation of relative mutation rates of mitochondrial DNA (mtDNA) and single-copy nuclear DNA (scnDNA) within lineages contributes to a general understanding of molecular evolutionary processes and facilitates making demographic inferences from population genetic data. The rate of divergence at synonymous sites ( K(s)) may be used as a surrogate for mutation rate. Such data are available for few organisms and no amphibians. Relative to mammals and birds, amphibian mtDNA is thought to evolve slowly, and the K(s) ratio of mtDNA to scnDNA would be expected to be low as well. Relative K(s) was estimated from a mitochondrial gene, ND2, and a nuclear gene, c-myc, using both "approximate" and likelihood methods. Three lineages of congeneric frogs were studied and this ratio was found to be approximately 16, the highest of previously reported ratios. No evidence of a low K(s) in the nuclear gene was found: c-myc codon usage was not biased, the K(s) was double the intron divergence rate, and the absolute K(s) was similar to estimates obtained here for other genes from other frog species. A high K(s) in mitochondrial vs. nuclear genes was unexpected in light of previous reports of a slow rate of mtDNA evolution in amphibians. These results highlight the need for further investigation of the effects of life history on mutation rates. PMID- 14745533 TI - Presence of dUTPase in the various human endogenous retrovirus K (HERV-K) families. AB - Various retroviruses have been shown to encode dUTPase. The overall phylogeny of dUTPase is unclear, though. The human genome contains a significant amount of human endogenous retroviruses (HERV) representing fossilized sequences of ancient exogenous retroviruses. A few HERV families have been reported to harbor dUTPase domains. We surveyed the various HERV families for the presence of dUTPase and found that ancestors of all HERV-K families but one encoded dUTPase. With two exceptions phylogenetic analysis shows a monophyletic origin of dUTPase for the different HERV-K dUTPases. Sequences of consensus dUTPase domains suggest that the various exogenous ancestors of HERV-K once encoded active enzymes. Our analysis provides informations on dUTPase phylogeny and further shows that endogenous retroviruses provide important informations regarding retrovirus evolution. PMID- 14745534 TI - Selective pressure on the allantoicase gene during vertebrate evolution. AB - During vertebrate evolution, the uric acid degradation pathway has been modified and several enzymes have been lost. Consequently, the end product of purine catabolism varies from species to species. In the past few years, we have focused our attention on vertebrate allantoicase (an uricolytic pathway enzyme), whose activity is present in certain fish and amphibians only, but whose mRNA we detected also in mammals. As allantoicase activity disappeared in amniotes, we wonder why these sequences not only remain present in the mammalian genome, but are still transcribed. To elucidate this issue, we have cloned and analyzed comparable cDNA sequences of different organisms from ascidians to mammals. The analysis of the nonsynonymous-synonymous substitution rate that we performed on the coding region comprising exons 3 to 8 by means of maximum likelihood suggested that a certain amount of purifying selection is acting on the allantoicase sequences. Some implications of the preservation of an apparently unnecessary gene in higher vertebrates are discussed. PMID- 14745535 TI - Highly variable polymorphism of the alpha-amylase gene family in Litopenaeus vannamei (Crustacea Decapoda). AB - Alpha-amylase from the tropical shrimp Litopenaeus vannamei presents a high degree of polymorphism and at least eight different electromorphs are detected by electrophoresis. Based on nucleotide sequences, three cDNAs have been previously characterized. In this paper we report on the organization and the evolution of corresponding alpha-amylase genes, determined after PCR amplification. Three AMY genes have been characterized, spanning over 3.3 kb and encoding mature proteins of 495 amino acids (aa), which are all expressed in the digestive gland. The existence of nine short introns, ranging from 86 to 454 bp, located at the same positions for each of the different genes, and presenting no similarity between them, is reported. Between 11 and 15% of changes are observed in the coding aa sequences of genes II and III compared to the gene I sequence respectively. One 5' putative promoter sequence has been sequenced and shows no classical TATA box upstream to the coding sequence. Based on the intron size difference, a single PCR (producing the S-R fragments) allows the separation of a partial gene I (750 bp), corresponding to cDNA 20, from the others (650-680 bp). Sequencing different S-R PCR fragments from one shrimp shows at least eight different haplotypes. A complex microsatellite repeat is present in intron 6 of gene II. Using size and sequence differences in this repeated portion, it is possible to characterize two gene subfamilies (IIa and IIb) encoding previously described cDNAs 28 and 37, respectively. For the gene II family, two to four alleles are present in one shrimp corresponding to these two genes. Within the Panama natural population, 35 different alleles are shown at this locus. Regarding alpha-amylase gene structure in the shrimp, many recombinants are present from a set of individuals and constitute an important mechanism of evolution of alpha-amylase function. PMID- 14745536 TI - Speciation in Chlamydia: genomewide phylogenetic analyses identified a reliable set of acquired genes. AB - Horizontal gene transfer (HGT), a process through which genomes acquire sequences from distantly related organisms, is believed to be a major source of genetic diversity in bacteria. A central question concerning the impact of HGT on bacterial genome evolution is the proportion of horizontally transferred sequences within genomes. This issue, however, remains unresolved because the various methods developed to detect potential HGT events identify different sets of genes. The present-day consensus is that phylogenetic analysis of individual genes is still the most objective and accurate approach for determining the occurrence and directionality of HGT. Here we present a genome-scale phylogenetic analysis of protein-encoding genes from five closely related Chlamydia, identifying a reliable set of sequences that have arisen via HGT since the divergence of the Chlamydia lineage. According to our knowledge, this is the first systematic phylogenetic inference-based attempt to establish a reliable set of acquired genes in a bacterial genome. Although Chlamydia are obligate intracellular parasites of higher eukaryotes, and thus suspected to be isolated from HGT more than the free-living species, our results show that their diversification has involved the introduction of foreign sequences into their genome. Furthermore, we also identified a complete set of genes that have undergone deletion, duplication, or rearrangement during this evolutionary period leading to the radiation of Chlamydia species. Our analysis may provide a deeper insight into how these medically important pathogens emerged and evolved from a common ancestor. PMID- 14745537 TI - A comparison of intraspecific patterns of DNA sequence variation in mitochondrial DNA, alpha-enolase, and MHC class II B loci in auklets (Charadriiformes: Alcidae). AB - Patterns of DNA sequence variation can be used to learn about mechanisms of organismal evolution, but only if mechanisms of sequence evolution are well understood. Although theories of molecular evolution are well developed, few empirical studies have addressed patterns and mechanisms of sequence evolution in nuclear genes within species. In the present study, we compared DNA sequences among three loci with different evolutionary constraints to determine the influences of effective population size, balancing selection, and linkage on intraspecific patterns of sequence variation. Specifically, we assessed the degree and nature of polymorphism in a 307-base pair (bp) fragment of the mitochondrial cytochrome b gene, intron VIII of the gene for alpha-enolase (a presumably neutral nuclear gene), and an approximately 600-bp fragment of an MHC class II B gene, including 155 bp of the hypervariable peptide binding region (a nuclear locus thought to be under balancing selection) for least and crested auklets (Aethia pusilla and A. cristatella; Charadriiformes: Alcidae). Transspecies polymorphism was found in both alpha-enolase and the MHC but not cytochrome b and, given estimates of effective population size, probably represents retained ancestral variation. Biases in nucleotide composition suggested that mutational bias, tRNA availability, and the secondary structure of mRNA and/or DNA may influence base usage. Several lines of evidence indicated that balancing selection may be acting on the MHC II B exon 2. However, no evidence of balancing selection was observed in the intron and exon sequences immediately downstream of MHC II B exon 2. PMID- 14745538 TI - Mammalian mutation pressure, synonymous codon choice, and mRNA degradation. AB - The usage of synonymous codons (SCs) in mammalian genes is highly correlated with local base composition and is therefore thought to be determined by mutation pressure. The usage is nonetheless structured. For instance, mammals share with Saccharomyces and Drosophila most preferences for the C-ending over the G-ending codon (or vice versa) within each fourfold-degenerate SC family and the fact that their SCs are placed along coding regions in ways that minimize the number of T|A and C|G dinucleotides ("|" being the codon boundary). TA and CG underrepresentations are observed everywhere in the mammalian genome affecting the SC usage, the amino acid composition of proteins, and the primary structure of introns and noncoding DNA. While the rarity of CG is ascribed to the high mutability of this dinucleotide, the rarity of TA in coding regions is considered adaptive because UA dinucleotides are cleaved by endoribonucleases. Here we present in vivo experimental evidence indicating that the number of T|A and/or C|G dinucleotides of a human gene can affect strongly the expression level and degradation of its mRNA. Our results are consistent with indirect evidence produced by other workers and with the detailed work that has been devoted to characterize UA cleavage in vitro and in vivo. We conclude that SC choice can influence strongly mRNA function and gene expression through effects not directly related to the codon-anticodon interaction. These effects should constrain heavily the nucleotide motif composition of the most abundant mRNAs in the transcriptome, in particular, their SC usage, a usage that must be reflected by cellular tRNA concentrations and thus defines for all other genes which SCs are translated fastest and most accurately. Furthermore, the need to avoid such effects genome-wide appears serious enough to have favored the evolution of biases in context-dependent mutation that reduce the occurrence of intrinsically unfavorable motifs, and/or, when possible, to have induced the molecular machinery mediating such effects to rely opportunistically on already existing motif rarities and abundances. This may explain why nucleotide motif preferences are very similar in transcribed and nontranscribed mammalian DNA even though the preferences appear to be adaptive only in transcribed DNA. PMID- 14745539 TI - Evolutionary history and mode of the amylase multigene family in Drosophila. AB - Previous studies indicate that the tandemly repeated members of the amylase (Amy) gene family evolved in a concerted manner in the melanogaster subgroup and in some other species. In this paper, we analyzed all of the 49 active and complete Amy gene sequences in Drosophila, mostly from subgenus Sophophora. Phylogenetic analysis indicated that the two types of diverged Amy genes in the Drosophila montium subgroup and Drosophila ananassae, which are located in distant chromosomal regions from each other, originated independently in different evolutionary lineages of the melanogaster group after the split of the obscura and melanogaster groups. One of the two clusters was lost after duplication in the melanogaster subgroup. Given the time, 24.9 mya, of divergence between the obscura and the melanogaster groups (Russo et al. 1995), the two duplication events were estimated to occur at about 13.96 +/- 1.93 and 12.38 +/- 1.76 mya in the montium subgroup and D. ananassae, respectively. An accelerated rate of amino acid changes was not observed in either lineage after these gene duplications. However, the G+C contents at the third codon positions (GC3) decreased significantly along one of the two Amy clusters both in the montium subgroup and in D. ananassae right after gene duplication. Furthermore, one of the two types of the Amy genes with a lower GC3 content has lost a specific regulatory element within the montium subgroup species and D. ananassae. While the tandemly repeated members evolved in a concerted manner, the two types of diverged Amy genes in Drosophila experienced frequent gene duplication, gene loss, and divergent evolution following the model of a birth-and-death process. PMID- 14745540 TI - Phylogeny and self-splicing ability of the plastid tRNA-Leu group I Intron. AB - Group I introns are mobile RNA enzymes (ribozymes) that encode conserved primary and secondary structures required for autocatalysis. The group I intron that interrupts the tRNA-Leu gene in cyanobacteria and plastids is remarkable because it is the oldest known intervening sequence and may have been present in the common ancestor of the cyanobacteria (i.e., 2.7-3.5 billion years old). This intron entered the eukaryotic domain through primary plastid endosymbiosis. We reconstructed the phylogeny of the tRNA-Leu intron and tested the in vitro self splicing ability of a diverse collection of these ribozymes to address the relationship between intron stability and autocatalysis. Our results suggest that the present-day intron distribution in plastids is best explained by strict vertical transmission, with no intron losses in land plants or a subset of the Stramenopiles (xanthophyceae/phaeophyceae) and frequent loss among green algae, as well as in the red algae and their secondary plastid derivatives (except the xanthophyceae/phaeophyceae lineage). Interestingly, all tested land plant introns could not self-splice in vitro and presumably have become dependent on a host factor to facilitate in vivo excision. The host dependence likely evolved once in the common ancestor of land plants. In all other plastid lineages, these ribozymes could either self-splice or complete only the first step of autocatalysis. PMID- 14745541 TI - The universal ancestor and the ancestor of bacteria were hyperthermophiles. AB - The definition of the node of the last universal common ancestor (LUCA) is justified in a topology of the unrooted universal tree. This definition allows previous analyses based on paralogous proteins to be extended to orthologous ones. In particular, the use of a thermophily index (based on the amino acids' propensity to enter the [hyper] thermophile proteins more frequently) and its correlation with the optimal growth temperature of the various organisms allow inferences to be made on the habitat in which the LUCA lived. The reconstruction of ancestral sequences by means of the maximum likelihood method and their attribution to the set of mesophilic or hyperthermophilic sequences have led to the following conclusions: the LUCA was a hyperthermophile "organism," as were the ancestors of the Archaea and Bacteria domains, while the ancestor of the Eukarya domain was a mesophile. These conclusions are independent of the presence of hyperthermophile bacteria in the sample of sequences used in the analysis and are therefore independent of whether or not these are the first lines of divergence in the Bacteria domain, as observed in the topology of the universal tree of ribosomal RNA. These conclusions are thus more easily understood under the hypothesis that the origin of life took place at a high temperature. PMID- 14745542 TI - Selection for highly biased amino acid frequency in the TolA cell envelope protein of Proteobacteria. AB - The bacterial cell envelope protein TolA functions to maintain the integrity of the cell membrane. This protein contains high levels of alanine and lysine that are used in the formation of alpha helices, which are required for normal protein function. The neutral model of molecular evolution predicts that amino acid composition and nucleotide composition are driven by the underlying GC content, as a result of mutation bias. However, this study shows that selection has acted to maintain high levels of alanine and lysine in the TolA protein of Proteobacteria, which in turn has biased nucleotide composition in the corresponding tolA gene. PMID- 14745543 TI - Lineage-specific homogenization of the polyubiquitin gene among human and great apes. AB - Ubiquitin is a highly conserved protein, and is encoded by a multigene family among eukaryote species. The polyubiquitin genes, UbB and UbC, comprise tandem multiple ubiquitin coding units without a spacer region or intron. We determined nucleotide sequences for the UbB and UbC of human, chimpanzee, gorilla, and orangutan. The ubiquitin repeat number of UbB was constant (3) in human and great apes, while that of UbC varied: 6 to 11 for human, 10 to 12 for chimpanzee, 8 for gorilla, and 10 for orangutan. The heterogeneity of the repeat number within closely related hominoid species suggests that a lineage-specific unequal crossover and/or gene duplication occurred. A marked homogenization of UbC occurred in gorilla with a low level of synonymous difference (p(s)). The homogenization of UbC also occurred in chimpanzee and less strikingly in human. The first and last ubiquitin coding units of UbC were clustered independently between species, and less affected by homogenization during the hominoid evolution. Therefore, the homogenization of ubiquitin coding units is likely due to an unequal crossing-over inside the ubiquitin units. The lineage-specific homogenization of UbC among closely related species suggests that concerted evolution has a key role in the short-term evolution of UbC. PMID- 14745546 TI - A simple immunometric assay to assess the feeding habits of Meprai spinolai, a Trypanosoma cruzi vector. AB - We propose a simple assay to assess the importance of seven vertebrate species as food sources for Mepria spinolai, a wild arthropod vector of Trypanosoma cruzi (the agent of Chagas' disease). Rabbits were immunized with serum proteins from one of each of the seven species. After titration, a consensus 1/100,000 dilution of the immune sera detected vertebrate serum proteins in the intestinal contents of 48.9% of 131 insects tested. The high proportion of negative samples is consistent with previous information indicating that these insects can withstand prolonged fasting periods. Alternatively, they may have fed on a different animal species than those used to produce the antisera. In about 70% of the positive samples, only one species of serum protein was detected. All pre-immune sera were negative. In 67% of the positive vectors, rabbit immunoglobulins were detected directly by means of a specific goat antibody. Thus, rabbits may play a role in T. cruzi transmission. PMID- 14745545 TI - Humoral responses against the C-terminal region of merozoite surface protein 1 can be remembered for more than 30 years in persons exposed to Plasmodium vivax. AB - Most people infected with Plasmodium vivax malaria developed antibodies against the C-terminal region of P. vivax merozoite surface protein (PvMSP1c) and the antibodies are sustained for a period up to 10 months after anti-malarial treatment. The longer-term stability of the specific humoral response was evaluated indirectly by determining the antibody titers in the sera from healthy individuals who lived an area from which malaria had been eradicated (450 persons) and an area in which it had recurred (1,524 persons). There were considerable residual antibody responses to PvMSP1c in over 15% of sera from healthy individuals, but only those who had lived in the era when malaria was prevalent. This means that antibodies against PvMSP1c may persist for more than 30 years, the malaria-free duration. This long-term memory of humoral immunity supports the C-terminal region of merozoite surface protein 1 as an effective malaria vaccine, in addition to the neutralizing activity reported previously. PMID- 14745547 TI - Differentiation of Babesia bigemina, B. bovis, B. divergens and B. major by Western blotting--first report of B. bovis in Austrian cattle. AB - To establish an assay for the serological differentiation of bovine Babesia species ( B. bigemina, B. bovis, B. divergens and B. major), antigens from experimentally infected cattle were Western blotted and probed with homologous and heterologous sera. Varying antigen patterns for each species allowed the determination of species-specific diagnostic antigens. Blood samples from 36 naturally infected cattle from the province of Styria were tested by indirect immunofluorescence antibody test (IFAT) against B. divergens, as well as by Western blotting against B. bigemina, B. bovis, B. divergens and B. major, 3 weeks after clinical babesiosis was diagnosed by blood smears. All 36 cattle were B. divergens-positive when tested by IFAT. In four cases (11%), an infection with both B. bovis and B. divergens and in two cases a single infection with B. bovis were diagnosed when tested by Western blot. B. bigemina and B. major infections were not detected. These are the first serologically confirmed cases of B. bovis in Austrian cattle. PMID- 14745548 TI - Dinurus ivanosi sp. nov. (Digenea: Hemiuridae) from the stomach of dolphin fish, Coryphaena hippurus, caught off the Kerala Coast, India. AB - A new species of stomach fluke, Dinurus ivanosi (sp. nov.), was recovered from the dolphin fish, Coryphaena hippurus, off the Kerala Coast, India. It is similar to other Dinurus species in the plicated prosoma, trilobed seminal vesicle and seven vitellaria, but differs from them in the length of the pharynx, unequal lobes of the seminal vesicle and unequal size of the testes. The most important diagnostic features of the new species are the general shape and proportions of the body, the position of the oral sucker, elongated pharynx, small distal lobe of the seminal vesicle, and larger anterior testis, smaller egg size, short and slender vitellaria and the limited extent of the vitelline field. PMID- 14745549 TI - Analysis of ataxia-telangiectasia mutated (ATM)- and Nijmegen breakage syndrome (NBS)-regulated gene expression patterns. AB - PURPOSE: Ataxia-telangiectasia (A-T) is a progressive, degenerative, complex autosomal recessive disease characterized by cerebellar degeneration, immunodeficiency, premature aging, radiosensitivity, and a predisposition to cancer. Mutations in the ataxia-telangiectasia mutated (atm) gene, which phosphorylates downstream effector proteins, are linked to A-T. One of the proteins phosphorylated by the ATM protein is Nijmegen Breakage Syndrome protein (NBS, p95/nibrin), which was recently shown to be encoded by a gene mutated in the Nijmegen breakage syndrome (nbs), an autosomal recessive disease with a phenotype virtually similar to that of A-T. The similarities in the clinical and cellular features of NBS and A-T have led us to hypothesize that the two corresponding gene products may function in similar ways in the cellular signaling pathway. Thus, we sought to identify genes whose expression is mediated by the atm and nbs gene products. MATERIAL AND METHODS: To identify genes, we performed an analysis of oligonucleotide microarrays using the appropriate cell lines, isogenic A-T (ATM-) and control cells (ATM+), and isogenic NBS (NBS-) and control cells (NBS+). RESULTS: We examined genes regulated by ATM and NBS, respectively. To determine the effect of ATM and NBS on gene expression in detail, we classified these genes into different functional categories, including those involved in apoptosis, cell cycle/DNA replication, growth/differentiation, signal transduction, cell-cell adhesion, and metabolism. In addition, we compared the genes regulated by the ATM and NBS to determine the relationship of their signaling pathways and to better understand their functional relationship. CONCLUSIONS: We found that, while ATM and NBS regulate several genes in common, both of these proteins also have distinct patterns of gene regulation, findings consistent with the functional overlap and distinctiveness of these two conditions. Due to the role of ATM and NBS in tumor suppression and the response to chemotherapy and radiotherapy, these findings may assist in the development of a more rational approach to cancer treatment, as well as a better understanding of tumorigenesis. PMID- 14745550 TI - Recombinant human erythropoietin attenuates weight loss in a murine cancer cachexia model. AB - BACKGROUND: Within hypoxic tumor regions anaerobic dissimilation of glucose is the sole source of energy generation. It yields only 5% of the ATP that is normally gained by means of oxidative glucose catabolism. The increased need for glucose may aggravate cancer cachexia. We investigated the impact of recombinant human erythropoietin (RhEPO) and increased inspiratory oxygen concentrations on weight loss in tumor-bearing mice. METHODS: Fragments of the murine C26-B adenocarcinoma were implanted in 60 BALB/c-mice. The mice were divided into four groups and assigned to: (A) no treatment; (B) RhEPO- administration (25 IU daily from day 1-11, three times per week from day 12); (C) RhEPO and 25% oxygen; and (D) RhEPO and 35% oxygen. Three control groups of four healthy mice each received the same treatment as groups A, B, and D, respectively. Hematocrit and hemoglobin levels, tumor volume, and body weight were monitored. At day 17 the experiment was terminated and the serum lactate concentration was measured. The tumors were excised and weighed and, for each mouse, the percentage weight loss was calculated. The impact of tumor weight and the treatments on lactate concentration and weight loss was evaluated. RESULTS: Significant positive correlations were found between tumor weight and lactate concentration and between tumor weight and percentage weight loss. In the mice with the largest tumors, RhEPO displayed a significant weight loss-reducing effect, and a significant negative correlation was found between hemoglobin concentration and weight loss. An oxygen-rich environment did not appear to influence weight loss. CONCLUSION: Anaerobic glycolysis in a growing C26-B tumor is related to weight loss. RhEPO administration results in a reduction of the percentage weight loss; this effect is probably mediated by an increased hemoglobin concentration. PMID- 14745551 TI - Gemcitabine-induced vasculitis in advanced transitional cell carcinoma of the bladder. AB - BACKGROUND: Gemcitabine (GEM) is an alternative chemotherapeutic agent for patients with metastatic bladder cancer. It is believed to be a well-balanced agent, having acceptable toxicity and enhanced antitumor activity. The integration of GEM into the initial chemotherapy plan for these patients is still being developed. CASE REPORT: The patient, male, aged 56 years, was suffering from a transitional cell carcinoma of the bladder. Due to frequent local superficial recurrences, radical cystectomy with pelvic lymphadenectomy and continent ileal diversion was performed. Four years after the operation a left inguinal lymphadenopathy was noted and metastatic bladder carcinoma was confirmed on biopsy. Cytotoxic therapy combining GEM and cisplatin and local external irradiation therapy was initiated. The patient developed extensive necrotising vasculitis with muscle damage after the second course of therapy. Chemotherapy was stopped immediately but this was not enough to relieve the symptoms of severe myalgia and swelling, and additional treatment consisting of cyclophosphamide and prednisolone was initiated. CONCLUSION: Although GEM seems to be relatively safe, some unexpected complications may occur during treatment. This case is not common, but it reinforces the need for careful attention to any new symptoms that seem to be unassociated with the primary disease. Prompt evaluation of such symptoms should be carried out in patients receiving GEM therapy. PMID- 14745552 TI - Paroxetine during breast-feeding: infant weight gain and maternal adherence to counsel. AB - The aims of the present study were to examine the weight gain in infants breast fed by mothers taking paroxetine and to assess the clinical implementation of the recommendations of the teratology service in our centre. A prospective cohort study design was used. The study group included 27 mothers who had taken paroxetine for at least 2 weeks while breast-feeding, 19 mothers who did not taken paroxetine and did not breast-feed their infants, and 27 mothers who breast fed their infants but did not take any drugs. Participants completed a detailed outcome questionnaire at 3 months to 1 year after the birth of their child. Additional infant data were obtained from specific follow-up forms routinely completed by the paediatricians and nurses at well baby clinics and paediatric services in Israel. Infant weight and developmental milestones at ages 3, 6, and 12 months were recorded. There were no statistically significant differences between the paroxetine group and control groups in mean infant weight at ages 6 and 12 months. The usual developmental milestones were reached in all groups. There were no reported adverse effects of paroxetine during lactation except for irritability in one infant. Mothers expressed satisfaction with the treatment. CONCLUSION: paroxetine use during breast-feeding does not affect infant weight gain and rarely has adverse effects on the infant. Paroxetine may be an acceptable solution for depression in lactating women provided that they adhere to four important restrictions: use lowest dose (20 mg/day), preferable single bed-time dose, avoid combinations of drugs, and ensure close medical follow-up of the infant. PMID- 14745553 TI - Acute renal failure after treatment with non-steroidal anti-inflammatory drugs. AB - Non-steroidal anti-inflammatory drugs (NSAIDs) are known to have adverse effects on kidney function. Situations with a stimulated renin-angiotensin system such as volume depletion or pre-existing chronic renal failure predispose to acute renal failure (ARF) via inhibition of prostaglandin synthesis by NSAIDs. To date, NSAIDs are frequently used as antipyretic drugs even in situations predisposing to ARF. Within 20 months, seven children presenting with diarrhoea and/or vomiting and fever were treated with therapeutic doses (11.5-32 mg/kg per day) of ibuprofen for 1 to 3 days before developing ARF. Maximum plasma creatinine levels were 180-650 micromol/l. One patient required emergency dialysis for hyperkalaemia, uraemia, and hyperphosphataemia. After cessation of NSAID treatment and rehydration, all patients recovered completely with a normalised creatinine level after 3 to 9 days. Once the acute phase is controlled, long-term outcome is excellent. Interstitial nephritis, another possible adverse effect of NSAIDs, might require steroid treatment and is the major differential diagnosis. Only histological examination can confirm the exact pathomechanism of ARF after NSAID exposure. If immunological events are responsible for the ARF, the recovery period is usually longer. CONCLUSION: non-steroidal anti-inflammatory drugs are potentially dangerous in situations with even moderate volume depletion. PMID- 14745554 TI - Malignant granular cell tumor of the esophagus. PMID- 14745555 TI - 21-kDa polypeptide, a low-molecular-weight cyclophilin, is released from pollen of higher plants into the extracellular medium in vitro. AB - Calcium ions play a key role in the elongation and orientation of pollen tubes. We found that significant amounts of 21-kDa polypeptide were specifically released into the extracellular medium when pollen grains of lily, Lilium longiflorum Thunb., were incubated in the presence of EGTA or at low concentrations of Ca2+. This phenomenon was also dependent on pH and on the concentrations of MgCl2 in the medium; the release of 21-kDa polypeptide from pollen was suppressed by increasing the MgCl2 concentration and by lowering pH. Germination of pollen grains was inhibited in the medium into which the 21-kDa polypeptide had been released. This inhibition was irreversible; germination did not occur on transfer of the pollen grains into basal culture medium. Immuno electron microscopy using an antibody against 21-kDa polypeptide showed that this polypeptide was present in the cytoplasm, vegetative nucleus and generative cell. When the pollen was treated with a medium containing EGTA, the density of 21-kDa polypeptide in the cytoplasm significantly decreased, but its density in vegetative nuclei and the generative cell did not, suggesting that only cytoplasmic 21-kDa polypeptide was released into the extracellular medium. The 21 kDa polypeptide was also present in the pollen of other higher-plant species, such as Tradescantia virginiana L., Nicotiana tabacum L. (angiosperms), and Cryptomeria japonica D. Don. (gymnosperm), and was also released into the medium in the presence of EGTA. In the case of C. japonica, however, it was released from pollen at alkaline pH above 8.5. The expression of 21-kDa polypeptide was not pollen-specific, because 21-kDa components immunoreactive with the anti-21 kDa polypeptide serum also existed in vegetative organs and cells of lily or tobacco. However, the 21-kDa polypeptide was not released into the extracellular medium from cultured tobacco BY-2 cells, even in the presence of EGTA. Amino acid sequences of two peptide fragments derived from 21-kDa polypeptide matched well those of low-molecular-weight cyclophilin (CyP). The antiserum against 21-kDa polypeptide recognized the CyP A from calf thymus and that in A431 carcinoma cells. The 21-kDa polypeptide fraction purified from lily pollen possessed peptidyl-prolyl cis- trans isomerase activity, which was suppressed by cyclosporin A (CsA), an inhibitor of enzyme activities of CyPs. From these results, we concluded that the 21-kDa polypeptide is a low-molecular-weight CyP. The present study showed that CyP in the pollen of higher plants is released into the extracellular matrix under unfavorable conditions. PMID- 14745556 TI - Formation of wheat ( Triticum aestivum L.) embryogenic callus involves peroxide generating germin-like oxalate oxidase. AB - In wheat ( Triticum aestivum L.), embryogenic callus formation comprises suppression of precocious germination by the zygotic embryo and the initiation of dedifferentiated cellular proliferation within it. Embryogenic calli are induced by treating immature embryos with 2,4-dichlorophenoxyacetic acid (2,4-D). Upon withdrawal from 2,4-D, somatic embryos develop from the periphery of the callus. Prior to visible callus formation, there is a striking induction of "germin-like" oxalate oxidase ("gl-OXO": EC 1.2.3.4) gene expression. Accumulation of gl-OXO mRNA is rapidly stimulated upon auxin treatment, with a consequent development of apoplastic enzyme activity producing H(2)O(2) within the cell wall. Within the dedifferentiated calli, gl-OXO enzyme activity becomes widespread over the surface of embryogenic calli. Differentiation of somatic embryos is initiated in regions of densely cytoplasmic, meristematic cells that are marked by highly localised expression of gl-OXO activity within these embryogenic cell masses. We suggest that this localised generation of H(2)O(2) by gl-OXO promotes peroxidative cross-linking of cell wall components, thereby preventing cellular expansion and maintaining these cell masses in an embryogenically competent condition. PMID- 14745557 TI - Incisional abdominal hernia: the open mesh repair. AB - BACKGROUND: Mesh techniques are the methods of choice for the repair of incisional hernias since these are due to the formation of unstable scar tissue. METHODS: We review the materials and techniques used in the repair of incisional hernias. We describe in detail the operative technique performed in our clinic, the pitfalls of the repair, and the overlap behind the xiphoid and the pubic bone. RESULTS: Polypropylene is the material widely used for open mesh repair. New developments have led to low-weight, large-pore polypropylene prostheses, which are adjusted to the physiological requirements of the abdominal wall and permit a proper tissue integration. These meshes provide the possibility of forming a scar net instead of a stiff scar plate and therefore help to avoid former known mesh complications. CONCLUSIONS: The ideal position for the mesh is the retromuscular sublay position where the force of the abdominal pressure holds the prosthesis against the deep surface of the muscles. The lowest incidence rates of recurrence have been reported for the retromuscular sublay repair; even after long-term follow-up recurrence rates of 10% are possible. Attaining such good results requires an adequate size of the mesh with sufficient overlap of at least 5-6 cm in all directions. Open mesh repair using modern low-weight polypropylene meshes in the retromuscular sublay technique offers excellent results for the treatment of incisional hernias. PMID- 14745558 TI - Paclitaxel and vinorelbine cause synergistic increases in apoptosis but not in microtubular disruption in human lung adenocarcinoma cells (A-549). AB - Concurrent administration of paclitaxel and vinorelbine results in cytotoxicity in vivo and in vitro in a number of tumor cell lines, yet the mechanisms of enhanced cell killing are undefined. In studies here, we show that low concentrations (1 nM) of paclitaxel and vinorelbine in combination result in enhanced cell killing by apoptosis ( P<0.05) in the human lung adenocarcinoma cell line, A-549. In contrast, necrotic cell death and formation of multinucleated cells, which were significantly increased by paclitaxel ( P<0.05) alone, but not vinorelbine, were not increased synergistically by both drugs. Paclitaxel also caused microtubular disruption which was not observed with vinorelbine. These data provide further rationale for the combined use of paclitaxel and vinorelbine in clinical trials, and suggest that the cooperative effects of drugs on apoptosis are not mediated through similar disruptional effects on microtubules. PMID- 14745559 TI - Cell transplantation in age related macular degeneration: current concepts and future hopes. PMID- 14745560 TI - Apoptotic cell death and microglial cell responses in cultured rat retina. AB - BACKGROUND: Optic nerve transection results in degeneration of axotomized retinal ganglion cells followed by the activation of resident microglial cells. METHODS: An organotypic culture of neonatal rat retina was used to examine the temporal aspect of retinal ganglion cell death and microglial cell recruitment. Retinas were fixed at various times after explantation and prepared for immunohistochemistry and lectin staining. RESULTS: Terminal deoxytransferase dUTP nick-end labeling (TUNEL) and immunohistochemistry for cleaved caspase-3 demonstrated a massive cascade of cell death in the ganglion cell layer (GCL) within hours after explantation. The rate of cell death in this layer was high and continued over a period of 48 h. In contrast, the rate of cell death was low in the outer nuclear layer (ONL) and apoptotic cells were evident after 6 days in vitro. Increases in the density of microglial cells in the GCL appeared to be recruited by proliferation within hours after explantation. In parallel, resident microglial cells also acquired an activated morphology as revealed by isolectin B(4) staining. Microglial cell activation in the GCL also included an upregulated expression for the lysosomal protein ED-1 and the cysteine protease inhibitor cystatin C. After 1 week of culture, immunolabeling for ED-1 demonstrated the presence of activated microglial cells also in the ONL. CONCLUSION: These data show rapid microglial cell recruitment and activation following the axotomy induced cell death of differentiated ganglion cells. The processes of microglial cell activation and cell death are slower in the outer retina. PMID- 14745561 TI - A retrospective study of preoperative CA 125 levels in 82 patients with ovarian cancer. AB - INTRODUCTION: The aim of our study was to investigate preoperative serum CA 125 as a prognostic factor in patients with ovarian carcinoma. METHODS: A retrospective analysis was conducted on 82 patients with ovarian carcinoma treated at our Unit between 1998 and 2000 who had a serum CA 125, evaluated by a commercially available radioimmunoassay, prior to cytoreductive surgery. We looked for an association between preoperative CA 125 and known prognostic factors of ovarian cancer. We compared outcomes of patients with preoperative CA 125 at or below to 500 U/ml with outcomes of patients with preoperative CA 125 above 500 U/ml. RESULTS: A significant ( p<0.002) correlation between stage and CA 125 serum levels was found as 16 out of 18 stage I-II patients (89%) had CA 125 level 500 U/ml. Among stage III and IV patients there was nonstatistically significant relation between serum CA 125 and histologic grade (G1+G2 vs. G3) and residual disease (<1 cm vs. >1 cm) after primary cytoreductive surgery. Preoperative serum CA-125 level did not predict either recurrences or disease free interval. CONCLUSION: Preoperative CA 125 correlated well with FIGO stage but not with age, grade, residual disease after primary surgery, relapse and disease free interval. PMID- 14745562 TI - Simultaneous supraventricular tachycardias in both fetuses of a twin gestation. AB - BACKGROUND: Fetal supraventricular tachycardia confers an increased risk of cardiac failure, hydrops, and eventual intrauterine death. Although protocols for prenatal anti-arrhythmic treatment are now well established, few published reports discuss this condition in the setting of multiple pregnancies. CASE REPORT: A 20-year-old primigravida woman with a twin pregnancy presented at 31 weeks of gestation for routine obstetrical check-up which revealed simultaneous supraventricular tachycardia in both fetuses. She was treated with oral digoxin, resulting in successful cardioversion in both of the fetuses, which was maintained until they were delivered by caesarian section at 38 weeks gestation. However, several hours after birth, tachyarrhythmias recurred in each of the infants. Combined disopyramide therapy with digoxin was necessary to control their heart rates. CONCLUSION: The treatment of arrhythmia in fetuses of a multiple gestation presents unique issues, particularly when diagnosed prior to fetal lung maturity. PMID- 14745563 TI - Tumor necrosis factor alpha secretion by peritoneal macrophages in patients with endometriosis. AB - INTRODUCTION: As a cytotoxic product of activated monocytes, macrophages, and lymphocytes, tumor necrosis factor alpha (TNF-alpha)--together with other cytokines and growth factors--is an important component in the immune response of the human organism. In addition, TNF-alpha plays a central role in neoangiogenesis. Because of its cytotoxicity with regard to several tumor cells and its motility-hindering effect on human sperm, TNF-alpha is considered to be a significant pelvic mediator of female sterility. OBJECTIVE: The goal of our study was to determine as to whether or not an increased TNF-alpha secretion by peritoneal macrophages (PM) can be measured in female patients with endometriosis compared with healthy subjects, and if TNF-alpha secretion can be correlated with the activity of endometriosis. METHODS: During infertility work-up, 100 female patients underwent a diagnostic laparoscopy. In accordance with the rAFS classification as well as from the macroscopic aspect of the degree of activity of the endometriosis, the patients were divided as follows: an endometriosis-free control group with a completely normal pelvic status (n=35) and three groups with increasing stages of endometriosis (n=65). In the control group (Group 1), the TNF-alpha concentrations (median values with minimum / maximum) were 6.2 pg/ml (1.9/10.2), in Group 2 with rAFS stage I/II less active endometriosis 56.33 pg/ml (39.5/71.2), in Group 3 with rAFS stage I/II but highly active endometriosis 81.41 pg/ml (68.4/98.7), while in Group 4 with rAFS stage III/IV 200,15 pg/ml (182.6/226.8), respectively. CONCLUSION: In conclusion, we were able to show that the TNF-alpha secretion of PM was significantly higher in patients with proven endometriosis compared to the control group. These results were found to be statistically significant and were in accordance with the histological findings. Thus, due to its immunomodulating potential, TNF-alpha may be a marker of both activity and stage of endometriosis. PMID- 14745564 TI - A survey of uterine perforation following dilatation and curettage or evacuation of retained products of conception. AB - STUDY: Of 11,914 women who underwent dilatation and curettage over a 7-year period (1995-2002) at Princess Badea Teaching Hospital in Irbid, North Jordan, 23 patients sustained a uterine perforation. In 22 cases, the operator was a trainee. Previous gynaecological surgery had been performed in only 2 of these 23 women. CONCLUSION: Operator inexperience seemed to be the only risk factor in this very common operation. PMID- 14745565 TI - The influence of vacuum extractor on fetal oxygenation and newborn status. AB - AIM: The aim of this study was to determine the effects of vacuum extractor assisted delivery on fetal oxygenation and acid-base balance. METHODS: Sixty-one women were enrolled in the present study. The subjects were divided into two groups. Group A, consisting of 39 women, had normal vaginal deliveries. Group B, consisting of 22 women, underwent a vacuum extractor assisted vaginal delivery. Fetal arterial oxygen saturation (SpO2) monitoring was used in all women after full cervical dilatation. After delivery, umbilical artery pH, pCO2, pO2 and base deficit (BDecf) levels were determined in all neonates. RESULTS: The mean FSpO2 value in Group A was 51.53+/-5.87% and in Group B 48.03+/-6.39% (p<0.03). The mean cord pH value in fetuses of Group A was 7.26+/-0.05, and in Group B 7.17+/ 0.09. There was also a significant difference between the two groups with regards to mean pO2, pCO2 and BDecf values. CONCLUSIONS: Vacuum assisted vaginal delivery was associated with lower fetal arterial oxygen saturation levels as well as lower cord blood pH values compared to those seen after unassisted vaginal delivery. Although decreased, however, the above parameters remained within normal ranges. PMID- 14745566 TI - Improved comfort in lower limb lengthening with the intramedullary skeletal kinetic distractor. Principles and preliminary clinical experiences. AB - INTRODUCTION: Limb lengthening by external fixators is associated with many problems such as pain at the pin tracts, muscle transfixation, pin tract infections, reduced joint motion, and prolonged fixation time. The intramedullary skeletal kinetic distractor (ISKD) is a new internal, mechanically activated implant, which distracts by mild rotations of 3 degrees. MATERIAL AND METHODS: In a prospective clinical study, four patients with an mean age of 29 years (18-36 years) underwent intramedullary lengthening via ISKD. The average lengthening of three femora and one tibia was 31 mm (26-40 mm). RESULTS: All patients performed the rotations for the distraction themselves without any significant problems. One patient took mild analgesics during the first days of distraction, whereas three patients did not require any analgesics. The average patient discharge occurred 10 days (8-11 days) postoperatively with no complications during the hospital stay. The planned length of distraction was achieved in all patients with normal alignment and normal joint orientation. Full weight bearing was performed on average after 10 weeks (7-14 weeks). Consolidation was noted 80 days (51-111 days) postoperatively with an average consolidation index of 2.9 days/mm. No complications were observed during the follow-up period of 14 months. The Enneking score was 26.8 points, and according to the classification of Paley all patients had an excellent result. CONCLUSIONS: From these preliminary results we conclude that the comfort of limb lengthening with the ISKD is increased by the elimination of fixator-associated complications and by the simple distraction mechanism, which is well tolerated by the patients. Further advantages of the ISKD are early full weight bearing and excellent limb function. PMID- 14745568 TI - The reliability of clinical examination in detecting pelvic fractures in blunt trauma patients: a meta-analysis. AB - INTRODUCTION: Several studies have recently questioned whether routine radiographic screening for pelvic fractures is necessary in the initial evaluation of blunt trauma patients. Therefore, we assessed how sensitive and specific the clinical examination is in detecting fractures of the pelvis. METHODS: We extensively searched various medical databases for studies that reported on the accuracy of pelvic examination in severely injured adults or children. Individual study results were summarized in a receiver operating characteristics (ROC) curve and pooled in a meta-analysis. RESULTS: Twelve studies with a total of 5454 patients met our inclusion criteria and provided data in sufficient detail. Pooled sensitivity and specificity were 0.90 (95% confidence interval: 0.85-0.93) and 0.90 (0.84-0.94), respectively. Results were better in those studies which excluded neurologically impaired patients [e.g., Glasgow Coma Scale (GCS) <13]. Among the 49 false negative cases whose fractures went undetected on clinical examination, the majority of patients had either altered consciousness or minor pelvic fracture only. Only 3 clinically relevant pelvic fractures were missed among 441 patients with fracture within a total population of 5235 patients. CONCLUSION: In stable and alert trauma patients, a thorough clinical examination will detect pelvic fractures with nearly 100% sensitivity, thus rendering initial radiography unnecessary in this group of patients. PMID- 14745567 TI - Comparative analysis of the benefits of autotransfusion of blood by a shed blood collector after total knee replacement. AB - INTRODUCTION: Joint replacement is associated with massive blood loss. Various techniques have been used to avoid the use of allogeneic blood. One of the techniques used is postoperative salvage and reinfusion of shed blood that was found to reduce the use of banked blood with its potential risk. MATERIALS AND METHODS: We prospectively studied 365 patients who underwent knee joint replacement (TKR) and were divided in two groups. Group A's shed blood (SureTrans System) was collected ( n=194) and reinfused and group B's was not ( n=171, "controls"). Hemoglobin levels before and after the operation were recorded. RESULTS: Allogeneic blood requirement for TKR decreased by 65% in group A compared to group B. The packed cell/patient index dropped from 0.91 to 0.29 in group 2A. Statistical analysis yielded the odds ratio for blood replacement, a "predicting formula" for blood replacement depending on hemoglobin levels, and a cutoff point for a patient's receiving blood replacement. CONCLUSION: We recommend using this system in TKR for decreasing allogeneic blood replacement and potential associated risks. The predicting formula for blood replacement may be a helpful tool when making a decision of whether or not to use the collector system and for whom. PMID- 14745569 TI - Age-related axonal and myelin changes in the rumpshaker mutation of the Plp gene. AB - The PLP1/Plp gene encodes proteolipid protein (PLP) and DM20, the major central nervous system myelin proteins. Mutations in the PLP1/ Plp gene cause dysmyelinating disorders in man and animals. The rumpshaker mutation was first identified in mice and later linked to a family diagnosed with neurological deficits akin to spastic paraplegia. The dysmyelination in the young rumpshaker mouse is well characterised. Here we report evidence for an age-related increase in myelin due mainly to the myelination of small axons, many large axons remain dysmyelinated. Levels of PLP/DM20 and myelin basic protein are considerably greater in myelin fractions from older compared with younger mutants. Myelin in sheaths of larger axons remains poorly compacted and may account for levels of 2',3'-cyclic nucleotide 3'-phosphodiesterase and myelin-associated glycoprotein being elevated over wild type in older mutant mice. A late-onset distal degeneration of the axons of the longest spinal tract, the fasciculus gracilis, is also noted. This is the first report of Wallerian-type degeneration in mice with spontaneous mutations of the Plp gene. PMID- 14745570 TI - Extrapyramidal features in patients with motor neuron disease and dementia; a clinicopathological correlative study. AB - Motor neuron disease (MND) may be complicated by frontotemporal dementia and/or an extrapyramidal movement disorder. Several studies have identified the pathological substrate for dementia in MND as being ubiquitin-immunoreactive inclusions and dystrophic neurites in the extramotor neocortex and hippocampus. Although degenerative changes have previously been noted in the basal ganglia and substantia nigra in MND, detailed pathological studies with clinical correlation are lacking. We examined postmortem material from eight patients with a history of MND and dementia, four of whom also had prominent extrapyramidal features. All cases showed the expected degenerative changes in the pyramidal motor system and ubiquitin-positive inclusions in the extramotor cortex. In addition, the cases with a history of extrapyramidal features had striking pathology in the basal ganglia and substantia nigra; neuronal loss and gliosis ranged from moderate to severe and immunohistochemistry demonstrated numerous neuronal inclusions and dystrophic neurites, which were reactive for ubiquitin, but not tau or alpha synuclein. Similar pathology was either absent or much milder in cases without extrapyramidal features. This study illustrates the utility of ubiquitin immunohistochemistry in demonstrating the range of pathology in MND and provides a neuropathological correlate for the extrapyramidal features which may occur in MND with dementia. PMID- 14745571 TI - Retinal neurodegeneration in the DBA/2J mouse-a model for ocular hypertension. AB - Mice of the DBA/2J strain spontaneously develop complex ocular abnormalities, including glaucomatous loss of retinal ganglion cells (RGC). In the present study ultrastructural features of retinal neurodegeneration in DBA/2J mice of different age (3, 6, 8 and 11 months) are described. By 3 months, RGC apoptosis characterized by electron-dense karioplasm and cytoplasm of ganglion cells was observed. The occurrence of apoptotic ganglion cells peaked at the age of 6 months. Past this age, necrosis characterized by swelling and electron-rare cytoplasm appeared to be the prevailing form of cell death. Muller glia activation increased with age, but there were no signs of leukocyte infiltration. At 8 and 11 months, signs of neoangiogenesis were found both at the ultrastructural level and in clinical examinations. In these older animals myelin like bodies, most probably representing the intracellular aggregates of phospholipids in irreversibly injured cells, were also seen. Photoreceptor cells were not affected at any age. Our observations suggest that retinal degeneration in the DBA/2J mice does not involve recruitment of blood-borne inflammatory/phagocytosing cells, and that apoptosis is gradually replaced by necrosis as the predominant pathway of RGC death. Retinal degeneration in 3- to 11-month-old DBA/2J mice partially resembles human pigment dispersion syndrome and pigmentary glaucoma with characteristic anterior segment changes and elevation of intraocular pressure. However, neovasculogenesis and myelin-like bodies are observed during aging. Therefore, the DBA/2J model requires judicious interpretation as a glaucoma model. PMID- 14745572 TI - Optimising the diagnostic value of anti-Saccharomyces cerevisiae-antibodies (ASCA) in Crohn's disease. AB - BACKGROUND/AIMS: Anti-Saccharomyces cerevisiae-antibodies (ASCA) are used to discriminate Crohn's disease (CD) from ulcerative colitis (UC). ASCA tests are not standardised and different methods for ASCA detection exist. This study was undertaken to compare ASCA tests and to clarify their diagnostic value. PATIENTS/METHODS: One hundred and two sera from CD patients, 53 from UC patients, and 50 sera from normal controls were examined for ASCA IgA and ASCA IgG using four different ELISA (Aesku.lab, Inova, Euroimmun, and Medipan) and indirect immunofluorescence (Euroimmun). Sensitivity, specificity, and positive and negative predictive values (PPV and NPV) for CD in this population were determined. Agreement between tests was expressed by kappa statistics. RESULTS/FINDINGS: The presence of either ASCA IgA or ASCA IgG had a PPV between 77 and 88%. Only the combined presence of ASCA IgA and ASCA IgG had a specificity which was constantly above 90%. Sensitivity, specificity, PPV, and NPV were then: Inova: 53, 97, 95, and 68%; Euroimmun ELISA: 46, 97, 94, and 66%; Aesku.lab: 50, 97, 94, and 66%; Medipan: 30, 98, 94, and 59%; indirect immunofluorescence: 51, 97, 94, and 66%. Agreement between ELISA test results of the Aesku.lab, Euroimmun, Medipan IgA and Inova systems was good (kappa: 0.63-0.79); between the Medipan IgG ELISA or indirect immunofluorescence and the others, it was lower (kappa: 0.33-0.6). If both ASCA IgA and ASCA IgG were detected by indirect immunofluorescence as well as ELISA, specificity for CD increased to >99% at a sensitivity of 23-38%. INTERPRETATION/CONCLUSIONS: The combined detection of ASCA IgA and ASCA IgG by indirect immunofluorescence as well as ELISA may optimise the discrimination of CD from UC. PMID- 14745574 TI - An audit of neonatal colostomy for high anorectal malformation: the developing world perspective. AB - A high divided sigmoid colostomy has been recommended for staged management of high anorectal malformation. We audited our cases of neonatal colostomy for high anorectal malformation to assess its effectiveness. A retrospective study was carried out of all surgical newborns admitted with high imperforate anus as the single diagnosis at our centre between December 1998 and December 2000. Morbidity and mortality were analysed after retrospective stratification into two groups (group A: birth weight >2.5 kg; group B: birth weight <2.5 kg). The chi square test was used to test the statistical significance in terms of outcome in the two groups. Overall mortality was 16%. Group A consisted of 34 babies: 30 with divided sigmoid colostomy and four with transverse loop colostomy. One baby with a divided sigmoid colostomy died from wound complications and septicaemia (mortality 2.9%). All four babies with transverse loop colostomy done under local anaesthesia survived, despite being sick on arrival. Group B consisted of 16 babies: 15 with sigmoid colostomy and one with transverse loop colostomy, with seven deaths (44%). None of the five babies with transverse loop colostomy done under local anaesthesia died, despite being sick on arrival, whereas all eight babies who died had undergone sigmoid colostomy under general anaesthesia. The difference in the outcomes of babies in groups A and B is highly significant ( p <.01). Sick, small (<2.5 kg) and septic babies arriving late to the unit do not appear to tolerate general anaesthesia and divided sigmoid colostomy well, despite that procedure's long-term advantages. Divided sigmoid colostomy has produced excellent results in babies >2.5 kg, but in the context of the developing world and limited critical care availability, transverse loop colostomy under local anaesthesia may save lives. PMID- 14745573 TI - Microarray-based gene expression profiling in pancreatic ductal carcinoma: status quo and perspectives. AB - BACKGROUND: Pancreatic ductal adenocarcinoma has an extremely poor prognosis. To improve the prognosis, novel molecular markers and targets for earlier diagnosis and adjuvant and/or neoadjuvant treatment need to be identified. One of the key techniques that has been developed to achieve this goal is DNA microarray profiling, which is used to identify the mechanisms of deregulated molecular functions in pancreatic carcinoma cells. OBJECTIVE: As several studies using microarrays have already been published, this review attempts to compare published data and crossvalidate the results. In addition, the applied techniques are discussed. PMID- 14745575 TI - Experience with primary laparoscopy-assisted endorectal pull-through for Hirschsprung's disease. AB - From October 1996 to July 2001, 61 patients received primary laparoscopy-assisted endorectal pull-through (LAERPT) for biopsy-proven Hirschsprung's disease (HD) at Mackay Memorial Hospital. The patients' ages at operation ranged from 7 days to 8 years. The patients were followed up for an average of 3.8 years (range 1-5.5 years). Major complications occurred in five (8.1%) patients, of whom four had surgical complications (two instances of anastomotic leakage, one colon perforation, and one delayed formation of colovesical fistula) and one had postoperative Salmonella infection-induced colonic stricture. The five patients required a diverting colostomy and a redo-pull-through procedure. Postoperative enterocolitis developed in 13 (21%) infants. All of them had enterocolitis before the diagnosis of HD was established. The majority of the episodes of postoperative enterocolitis were mild and resolved spontaneously, but four neonates were hospitalized with significant systemic manifestations, and two of them needed a second operation to solve the problem. After primary LAERPT, stooling frequency in young infants declined rapidly from 10.5+/-3.2 to 4.4+/-1.6 bowel movements per day in the first 3 months and more slowly thereafter. Most of these infants had regular bowel movements one to two times per day 1 year after operation. Continence evaluation of the 43 patients over 3 years of age was graded as normal in 24 (56%) patients and good in 19 (44%) patients. Of the 13 patients between 1 and 3 years of age, 11 (85%) had regular normal stools, and two (15%) had occasional soiling. From the current study, the authors concluded that primary LAERPT is a safe and effective method of managing HD with excellent continence results. Considering the complications of surgery and postoperative enterocolitis, patients with long segment aganglionosis, severe enterocolitis, or prominently dilated colon are not good candidates for primary LAERPT. PMID- 14745576 TI - Diagnostic laparoscopy with planned appendectomy: an integral step in the evaluation of unexplained right lower quadrant pain. AB - Recurrent right lower quadrant (RLQ) abdominal pain is often difficult to diagnose and treat. We reviewed our experience with diagnostic laparoscopy with planned appendectomy for children with unexplained RLQ pain. This procedure was performed when the cause of atypical RLQ pain could not be diagnosed by comprehensive medical and radiological evaluation. Outcome data was obtained at office visits and by telephone. From 1997 to 2000, 30 children (22 female) presented with unexplained RLQ pain. Ages ranged from 5 to 16 years (mean 11 years). Symptoms had been present from 6 days to 2 years (median 6 weeks). Gross and/or histological appendiceal abnormalities were found in 26 children (87%). Incidental findings included patent processus vaginalis in one and adnexal cysts in six. Two complications occurred: pelvic fluid collection and umbilical suture reaction. At initial follow-up, 29 patients (97%) were pain-free. Long-term follow-up was continued through 2002. At a median of 19 months (range 2-47 months), 25 of 28 patients (89%) reported no recurrence of their original pain. Our results confirm that the appendix is an important source of unexplained recurrent RLQ pain in children. Diagnostic laparoscopy with planned appendectomy is highly effective and should be considered an integral step in the management of these patients. PMID- 14745577 TI - Posterior urethral valves: incidence and progress of vesicoureteric reflux after primary fulguration. AB - This is a prospective study of 20 cases of posterior urethral valves (PUV) presenting between the ages of 12 days and 5.5 years (median 15 months) in order to determine the incidence and progress of vesicoureteric reflux (VUR), hydronephrosis (HN), and renal functional status. The efficacy of fulguration as the sole modality of treatment for PUV was assessed in terms of improvement or disappearance of VUR over a 6-month follow-up period. VUR was present in 60% of the patients, being unilateral in 41.7%. Out of 19 renal units with VUR, reflux subsided in 31.5% by 3 months and 78.94% by 6 months. The blood urea and serum creatinine levels, which were raised in 50% of the patients at presentation, came down to normal by 6 months in all the cases. Improvement in glomerular filtration rates (GFR) was noted in all the children at each follow-up and was found to be statistically significant ( p<0.01). HN was present in all the patients at presentation and was bilateral in 90%. It decreased significantly during the follow-up period, though its complete disappearance was seen only in one case. Vesicoureteric reflux dysplasia syndrome (VURD) was present in two cases. Our study showed that VUR disappeared in a majority of the cases by 6 months once adequate urethral patency was restored, although hydronephrosis persisted. PMID- 14745578 TI - Eyebrow incision with supraorbital trephination for endoscopic corpus callosotomy: a feasibility study. AB - OBJECTIVE: We describe a novel technique for the partial bisection of the corpus callosum in order to increase the minimally invasiveness of this procedure. METHODS: Brow incisions with midline trephinations were performed in six adult cadavers. An endoscope was next introduced and used to transect approximately the anterior two-thirds of the corpus callosum. RESULTS: No complications such as injury to the superior sagittal sinus or anterior cerebral artery were encountered in any of our cadaveric specimens. The corpus callosum was easily transected in each specimen. CONCLUSION: As a feasibility study, we believe this technique could provide a less invasive mechanism for patients who require corpus callosotomy and will minimize much of the morbidity associated with the traditional methods of sectioning the corpus callosum. PMID- 14745580 TI - Power and control muscles of cicada song: structural and contractile heterogeneity. AB - Sound production in cicadas is powered by a pair of large muscles whose contractions cause buckling of cuticular tymbals and thereby create sound pulses. Sound is modulated by control muscles that alter the stiffness of the tymbals or change the shape of the abdominal resonance chamber. Muscle ultrastructure and contractile properties were characterized for the tymbal muscle and two control muscles, the ventral longitudinal muscle and the tymbal tensor, of the periodical cicada Magicicada septendecim. The tymbal muscle is a fast muscle that is innervated by a single motoraxon. The control muscles are an order of magnitude less massive than the tymbal muscles, but their innervation patterns were considerably more complex. The tensor muscle is innervated by two axons, each of which evokes rather slow twitches, and the ventral muscle is innervated by at least six axons, some of which produce fast and the others slow contractions. Muscle contraction kinetics correlated well with ultrastructure. Fibers of the tymbal muscle and the portions of the ventral muscle thought to be fast were richly supplied with transverse tubules (T-tubules) and sarcoplasmic reticulum (SR); slow portions of the ventral muscle and the tensor muscle had relatively little SR. PMID- 14745579 TI - Split cord malformation and situs inversus totalis: case report and review of the literature. AB - INTRODUCTION: Situs inversus is a rare condition of visceral transposition in which the spinal axis is rarely affected. CASE REPORT: The authors report a patient with situs inversus totalis and Type II split cord malformation. The patient had no symptoms and presented with scoliosis. CONCLUSIONS: Recent compelling evidence from animal models and human case reports has lead to hypotheses that defects of the midline and laterality defects (e.g., situs inversus) are etiologically related. Confirmation from additional case reports of situs inversus and split cord malformation could prove useful in determining a genetic locus for split cord malformations or implicating various chemical agents that are known to produce situs inversus as potential causative factors in the production of split cord malformations. PMID- 14745581 TI - Ovarian maldescent. AB - Undescended ovary is a rare disorder that can be associated with mullerian malformations. There is an unclear association with infertility and malignant disease. When an ovary is not in its normal location, it is sought in other locations above the pelvic brim. This is important in cases of undetermined cyclical abdominal pain, follicle aspiration and surgical castration. MRI has proven to be the best imaging method for finding an ovary in an anomalous position and for disregarding associated malformations. We report two patients with unilateral ovarian maldescent, one of them with Rokitansky-Kuster-Hauser syndrome. A review of the literature is included. PMID- 14745582 TI - Production of an endoglucanase by the shipworm bacterium, Teredinobacter turnirae. AB - The nutritional behavior of a cellulolytic nitrogen-fixing shipworm bacterium, Teredinobacter turnirae, is described, with respect to various carbon and nitrogen sources, in terms of endoglucanase production. Also, the effects of various surfactants on enzyme production are reported. Among the carbon sources, sucrose results in the maximum enzyme production, followed by cellulose. Ammonium phosphate proves to be the best nitrogen source for endoglucanase production. Various surfactants enhance the enzyme titers, with Triton X-100 yielding the best results. A combination of the above-mentioned components improves the enzyme production by 3.6-fold. PMID- 14745583 TI - Molecular phylogeny of monocotyledons inferred from combined analysis of plastid matK and rbcL gene sequences. AB - Using matK and rbcL sequences (3,269 bp in total) from 113 genera of 45 families, we conducted a combined analysis to contribute to the understanding of major evolutionary relationships in the monocotyledons. Trees resulting from the parsimony analysis are similar to those generated by earlier single or multiple gene analyses, but their strict consensus tree provides much better resolution of relationships among major clades. We find that Acorus (Acorales) is a sister group to the rest of the monocots, which receives 100% bootstrap support. A clade comprising Alismatales is diverged as the next branch, followed successively by Petrosaviaceae, the Dioscoreales-Pandanales clade, Liliales, Asparagales and commelinoids. All of these clades are strongly supported (with more than 90% bootstrap support). The sister-group relationship is also strongly supported between Alismatales and the remaining monocots (except for Acorus) (100%), between Petrosaviaceae and the remaining monocots (except for Acorus and Alismatales) (100%), between the clade comprising Dioscoreales and Pandanales and the clade comprising Liliales, Asparagales and commelinoids (87%), and between Liliales and the Asparagales-commelinoids clade (89%). Only the sister-group relationship between Asparagales and commelinoids is weakly supported (68%). Results also support the inclusion of Petrosaviaceae in its own order Petrosaviales, Nartheciaceae in Dioscoreales and Hanguanaceae in Commelinales. PMID- 14745584 TI - How do rhesus monkeys ( Macaca mulatta) scan faces in a visual paired comparison task? AB - When novel and familiar faces are viewed simultaneously, humans and monkeys show a preference for looking at the novel face. The facial features attended to in familiar and novel faces, were determined by analyzing the visual exploration patterns, or scanpaths, of four monkeys performing a visual paired comparison task. In this task, the viewer was first familiarized with an image and then it was presented simultaneously with a novel and the familiar image. A looking preference for the novel image indicated that the viewer recognized the familiar image and hence differentiates between the familiar and the novel images. Scanpaths and relative looking preference were compared for four types of images: (1) familiar and novel objects, (2) familiar and novel monkey faces with neutral expressions, (3) familiar and novel inverted monkey faces, and (4) faces from the same monkey with different facial expressions. Looking time was significantly longer for the novel face, whether it was neutral, expressing an emotion, or inverted. Monkeys did not show a preference, or an aversion, for looking at aggressive or affiliative facial expressions. The analysis of scanpaths indicated that the eyes were the most explored facial feature in all faces. When faces expressed emotions such as a fear grimace, then monkeys scanned features of the face, which contributed to the uniqueness of the expression. Inverted facial images were scanned similarly to upright images. Precise measurement of eye movements during the visual paired comparison task, allowed a novel and more quantitative assessment of the perceptual processes involved the spontaneous visual exploration of faces and facial expressions. These studies indicate that non-human primates carry out the visual analysis of complex images such as faces in a characteristic and quantifiable manner. PMID- 14745585 TI - Genetic and environmental factors in the pathogenesis of Huntington's disease. AB - Huntington's disease is a fatal inherited disorder in which there is progressive neurodegeneration in specific brain areas, mainly the striatum and cerebral cortex, producing motor, cognitive, and psychiatric symptoms. The trinucleotide repeat mutation involved is common to many other brain diseases, which may therefore involve similar mechanisms of pathogenesis. We are beginning to understand how a CAG trinucleotide repeat expansion in the disease gene, encoding an expanded polyglutamine tract, induces neuronal dysfunction and symptomatology in Huntington's disease. Recent evidence that environmental factors modify the onset and progression of neurodegeneration has shed new light on Huntington's disease and other devastating brain diseases. This review focuses on genetic mediators, environmental modulators, and associated gene-environment interactions in the pathogenesis of Huntington's disease. PMID- 14745586 TI - Importance of statistical significance. PMID- 14745587 TI - Historical comment on modern work. PMID- 14745589 TI - Acquired indirect inguinal hernias. PMID- 14745590 TI - Relationship between dental erosion, soft drink consumption, and gastroesophageal reflux among Icelanders. AB - The aim of this study was to determine the prevalence of dental erosion in young Icelandic adults (19-22 years old) and patients with gastroesophageal reflux disease (GERD), in relation to their soft drink consumption and gastroesophageal reflux. Eighty subjects (40 males and 40 females), comprising 57 young adults (mean age 21 +/- 2 years) and 23 GERD patients (mean age 35 +/- 10 years), were enrolled in this study. All subjects answered a detailed frequency questionnaire of soft drink consumption and participated in a clinical examination. Erosion was scored for incisor and molar teeth separately. No significant difference was observed in the prevalence of dental erosion between young adults and GERD patients. However, by combining the two study groups a three-fold higher risk of having erosion in molars or incisors was found for subjects drinking Coca-Cola three times a week or more often ( p < 0.05). Additionally, significantly higher erosion scores were found in molars among subjects drinking more than 1 litre of carbonated drinks (all brands) per week ( p < 0.05). It is concluded that the frequency of soft drink consumption is a strong risk factor in the development of dental erosion. PMID- 14745591 TI - Identification of a collagen-like protein gene from white spot syndrome virus. AB - The most unique feature of white spot syndrome virus (WSSV) is the presence of a collagen-like protein (termed as WSSV-CLP). In this report, the N-terminal fragment of WSSV-CLP (CLPn) was expressed as a fusion protein with glutathione S transferase (GST) in Escherichia coli and purified. Specific antibody was then raised against the purified fusion protein (GST-CLPn). Temporal analysis showed that the WSSV collagen gene was an early viral gene. Immunogold localization using specific antibody revealed that the gold particles, under a transmission electron microscope, were presented along the outer envelope of WSSV virions. This experiment suggested that the collagen gene encoded an envelope protein of WSSV. Using immuno-affinity chromatography, the WSSV-CLP was purified from crudely purified WSSV virions. The WSSV-CLP was N-glycosylated, as indicated by the increased migration in SDS-PAGE after treatment with N-linked glycosidase F. PMID- 14745592 TI - Further characterization of 'sweet potato virus 2': a distinct species of the genus Potyvirus. AB - An incompletely described potyvirus isolate from sweet potato in Taiwan, referred to as 'sweet potato virus 2' (SPV2), was further characterised. Electron microscopy revealed that SPV2 has filamentous particles of 850 nm in length and induces cytoplasmic cylindrical inclusions consisting of pinwheels and scrolls. The virus was mechanically transmitted to several species of the genera Chenopodium, Datura, Nicotiana, and Ipomoea. Two biotypes of Myzus persicae transmitted SPV2 in a non-persistent manner. Decoration titer experiments revealed a distant serological relationship between SPV2 and other potyviruses infecting sweet potato. The 3'-terminal 2006 nucleotides of the viral RNA were determined and shown to be a potyviral genome fragment comprising the coding region for the C-terminal half of the NIb protein, the entire coat protein cistron, and the 3' untranslated region (UTR). Comparison of the capsid protein and 3' UTR sequences of SPV2 with those of other potyviruses demonstrated that it is a distinct member of the genus Potyvirus (family Potyviridae). We propose that SPV2 is named Sweet potato virus Y. PMID- 14745593 TI - Characterization of Plys-proximal morphogenetic genes of transposable bacteriophage Mu. AB - Late during the bacteriophage Mu lytic cycle, Mu DNA must be matured and packaged from its dispersed integration sites in the host DNA in order to produce progeny virions. Whereas control of late gene transcription in Mu is becoming well understood, less is known about the phage morphogenetic process. To investigate the latter, we cloned and sequenced a approximately 4.3-kb region of the phage DNA beginning just upstream of the leftmost late promoter Plys. Previous mapping of amber mutations had located the lysis (lys) and proposed DNA maturation genes D and E in this region. When the DNA sequence was analyzed, seven potential open reading frames were found. DNA sequence analysis of amber mutations in genes D and E identified the sixth and seventh open reading frames as D and E, respectively. Cloning and expression of this region enabled production of cell free protein extracts that specifically recognize the phage-encoded packaging sequence (pac), a characteristic exhibited by phage maturation enzymes. In addition, the E protein was found to share homology with the large subunit of many phage DNA maturation enzymes. These results support the hypothesis that D and E encode subunits of the Mu DNA maturation enzyme. PMID- 14745594 TI - Unambiguous identification of JC polyomavirus strains transmitted from parents to children. AB - JC polyomavirus (JCV), the etiological agent of progressive multifocal leukoencephalopathy, is ubiquitous in humans, infecting children asymptomatically, then persisting in renal tissue. It has been proposed that JCV is transmitted mainly from parents to children through long-term cohabitation. The objective of this study was to further elucidate the mode of JCV transmission. In 5 families, we selected parent/child pairs between whom JCV was probably transmitted (judged on the basis of the identity of a 610-bp JCV DNA sequence between the parent and child). We established 5 to 9 complete JCV DNA clones from the urine of each parent or child. The complete sequences of these clones were determined and compared in each family. Nucleotide substitutions were detected in 4 parents and 1 child, and sequence rearrangements (deletions or duplications) were found in 2 parents and 2 children. Phylogenetic comparison of the detected sequences indicated that the diversity of JCV DNA sequences was generated in each family (i.e. not caused by multiple infection). We found that in 4 of the 5 families, a sequence detected in the parent was completely identical to one in the child. These findings provided further support for the proposed mode of JCV transmission, i.e. parent-to-child transmission during cohabitation. PMID- 14745595 TI - Biochemical properties of the P42 protein encoded by RNA segment 6 of influenza C virus. AB - P42, encoded by a colinear transcript of Influenza C virus RNA segment 6 (M gene), is an integral membrane protein which is cleaved by signal peptidase to generate M1' and CM2 composed of N-terminal 259 amino acids and C-terminal 115 amino acids, respectively. Herein, the biochemical features of P42 were investigated. N-glycosylated form of P42, designated P44, forms disulphide-linked dimers and tetramers. P44 is transported to the Golgi apparatus, but not to the trans-Golgi, since P44 is completely sensitive to endoglycosidase H. P44 and P42 are unstable irrespective of N-glycosylation or oligomerization. 26S proteasome inhibitor, lactacystin prevented the degradation of P42 as well as M1', but not that of P44 efficiently, suggesting that P44 is degraded by another protease besides the 26S proteasome. PMID- 14745596 TI - Epstein-Barr virus-mediated protection against etoposide-induced apoptosis in BJA B B cell lymphoma cells: role of Bcl-2 and caspase proteins. AB - Epstein-Barr virus (EBV)-infected B cell lymphomas are resistant to apoptosis during cancer development and treatment with therapies. The molecular controls that determine why EBV infection causes apoptosis resistance need further definition. EBV-positive and EBV-negative BJA-B B cell lymphoma cell lines were used to compare the expression of selected apoptosis-regulating Bcl-2 and caspase proteins in EBV-related apoptosis resistance, after 8 hr or 18-24 hr etoposide treatment (80 microM). Apoptosis was quantified using morphology and verified with Hoechst 33258 nuclear stain and electron microscopy. Fluorescence activated cell sorting (FACS) was used to analyse effects on cell cycle of the EBV infection as well as etoposide treatment. Anti-apoptotic Bcl-2 and Bcl-XL, pro apoptotic Bax, caspase-3 and caspase-9 expression and activation were analysed using Western immunoblots and densitometry. EBV-positive cultures had significantly lower levels of apoptosis in untreated and etoposide-treated cultures in comparison with EBV-negative cultures (p < 0.05). FACS analysis indicated a strong G2/M block in both cell sublines after etoposide treatment. Endogenous Bcl-2 was minimal in the EBV-negative cells in comparison with strong expression in EBV-positive cells. These levels did not alter with etoposide treatment. Bcl-XL was expressed endogenously in both cell lines and had reduced expression in EBV-negative cells after etoposide treatment. Bax showed no etoposide-induced alterations in expression. Pro-caspase-9 and -3 were seen in both EBV-positive and -negative cells. Etoposide induced cleavage of caspase-9 in both cell lines, with the EBV-positive cells having proportionally less cleavage product, in agreement with their lower levels of apoptosis. Caspase-3 cleavage occurred in the EBV-negative etoposide-treated cells but not in the EBV-positive cells. The results indicate that apoptosis resistance in EBV-infected B cell lymphomas is promoted by an inactive caspase-3 pathway and elevated expression of Bcl-2 that is not altered by etoposide drug treatment. PMID- 14745597 TI - Translation enhancer in the 3'-untranslated region of rotavirus gene 6 mRNA promotes expression of the major capsid protein VP6. AB - The eleven rotavirus mRNAs contain 5'-cap structures and most end with the 3' consensus sequence 5'-UGACC-3'. The UGACC functions as a common translation enhancer (3'-TE-con) that upregulates viral protein expression through a process mediated by the nonstructural protein NSP3. To address the possibility that gene specific enhancers are also contained in the untranslated regions (UTRs) of the rotavirus mRNAs, we used rabbit reticulocyte lysates to investigate the translation efficiencies of analog RNAs containing viral-specific 5'-and 3'-UTRs and the open reading frame for chloramphenicol acetyltransferase. These experiments combined with the analysis of full-length viral RNAs and RNAs containing 3'-truncations showed that a highly active enhancer was present near the 5'-end of the 139-nucleotide 3'-UTR of the gene 6 mRNA (3'-TEg6). The 3'-TEg6 represents a functionally independent enhancer, as no other portion of the gene 6 mRNA was required for its activity. The 3'-TEg6 differs significantly from the 3' TE-con in that the gene 6-specific enhancer does not require viral protein for activity and is formed by a sequence unique to only one of the eleven viral mRNAs. Together, our findings suggest that the 3'-UTR of the gene 6 mRNA contains two TEs, one is gene-specific (3'-TEg6) and the other is common to nearly all rotavirus genes (3'-TE-con). The activity of the 3'-TEg6 is likely important for directing the efficient translation of the gene 6 mRNA at levels sufficient to provide the 780 copies of VP6 necessary for the assembly of each progeny virion. PMID- 14745599 TI - Sequencing and characterization of the full-length gene encoding the single stranded DNA binding protein of a novel Chelonian herpesvirus. AB - Through 4 consecutive genomic walks employing a recently modified inverse polymerase chain reaction technique, a 4,054-bp DNA fragment of a newfound green turtle herpesvirus (GTHV) was obtained from tumor tissues of a green turtle with fibropapillomas. This newly identified viral DNA fragment contains two non overlapping open reading frames (ORF) oriented in the 3' to 5' direction. The first ORF is 59% G+C rich and contains the full-length genomic sequence of the DNA binding protein (DBP) gene (3,585 bp) encoding a protein of 1,195 amino acid residues in length. The second ORF encodes a partial peptide of the UL28 gene. Phylogenetic analysis of the GTHV DBP gene confirmed and substantiated that this novel Chelonian herpesvirus is closely related to the subfamily Alphaherpesvirinae. Examination of the translated amino acid sequence further supports this categorization since GTHV DBP comprises a highly conserved zinc finger motif (CXLCX4RX2C) and a putative DNA binding domain, and exhibits high sequence homology to other alphaherpesviruses. Cloning and sequencing the genome of this putative herpesvirus will facilitate current understanding of its role in causing GTFP and the development of molecular- and immuno-based methods for the diagnosis and prevention of this devastating disease of green turtles. PMID- 14745598 TI - HCV core, NS3, NS5A and NS5B proteins modulate cell proliferation independently from p53 expression in hepatocarcinoma cell lines. AB - Several reports have shown that activity and/or expression of p53 can be modulated by Hepatitis C virus (HCV) proteins and may interfere with normal regulation of cell growth. In order to understand the relationship between p53 function and HCV proteins expression, we have investigated potential effects of the core, NS3, NS5A and NS5B proteins on Huh-7 (p53 +/+) and Hep3B (p53 -/-) cell proliferation. The effect of HCV proteins transiently expressed after recombinant adenoviral infection was analyzed by Western blot, crystal violet and propidium iodide staining. Expression of the core, NS3, NS5A or NS5B proteins inhibited cell proliferation and blocked both cell lines in the G2/M phase of the cell cycle. c-myc and p53 expression were respectively induced and increased in Huh-7 cells only following expression of the Core protein. No expression of p21(waf1/cip1) could be detected and expression of cyclin A, cdk2 and p27(Kip1) were independent of HCV protein's expression. Our results show that the effect of core, NS3, NS5A and NS5B on cell proliferation is independent of p53 expression and that only the Core protein, induces the expression of both c-myc and p53. PMID- 14745600 TI - A virus related to soybean mosaic virus from Pinellia ternata in China and its comparison with local soybean SMV isolates. AB - A potyvirus isolated from Pinellia ternata in China was characterised and shown to be related to Soybean mosaic virus (SMV). The virus was pathogenic on P. ternata and some soybean cultivars, whereas the local soybean SMV isolate HH5 did not infect P. ternata. Western blot experiments demonstrated a serological relationship between the virus from Pinellia, SMV and Watermelon mosaic virus (WMV). The complete nucleotide sequences of the Pinellia virus (isolate P-1, 9735 nt) and of the Chinese soybean SMV isolates HH5 (9585 nt) and HZ (9588 nt) were determined. A 1733 nt sequence at the 3'-terminus of a second isolate from Pinellia (isolate P-2) was also determined. The predicted polyprotein of isolate P-1 has 83% amino acid (aa) identity with those of published SMV sequences. In many parts of the genome, aa identity was about 90% but it was much lower in the P1 protein region (24-29%), where it more closely resembled Dasheen mosaic virus (62%). The partial sequence of isolate P-2 had 91% nt identity to P-1 and both isolates resembled a recent sequence in the public databases (AF469171) wrongly named Zantedeschia mosaic virus. The two complete SMV soybean sequences had 93 95% nt identity with those of the previously sequenced isolates and >97% amino acid identity. Phylogenetic analysis and comparisons of coat proteins suggest that the Pinellia, WMV and SMV potyviruses should probably be treated as strains of the same species. PMID- 14745601 TI - Expression of a foot-and-mouth disease virus immunodominant epitope by a filamentous bacteriophage vector. AB - We described the construction of a recombinant filamentous phage displaying on its surface the immunodominant site of VP1 protein of foot-and-mouth disease virus (FMDV). The coding sequence was inserted at the amino-terminus of the major coat protein pVIII via a spacer. The hybrid phage proved to be antigenic as it was recognized by polyclonal and monoclonal anti FMDV sera. In two experiments involving immunisation of guinea-pigs with the recombinant phage, a low antibody response was generated. This suggests a possible role for phage displayed peptides in inducing anti FMDV immunity and the possibility of further development is discussed. PMID- 14745602 TI - Breakage of resistance to Cucumber mosaic virus by co-infection with Zucchini yellow mosaic virus: enhancement of CMV accumulation independent of symptom expression. AB - Resistance to the cucumovirus Cucumber mosaic virus (CMV) in cucumber cv. Delila was manifested as a very low level of accumulation of viral RNA and capsid protein, and an absence of CMV-induced symptoms. In addition, resistance was observed at the single cell level, with a reduction in accumulation of CMV RNAs, compared to accumulation in cells of the susceptible cucumber cv. Bet Alpha. Resistance to CMV in cv. Delila was broken by co-infection with the potyvirus Zucchini yellow mosaic virus (ZYMV). Resistance breakage in cv. Delila plants was manifested by an increase in the accumulation of (+) and (-) CMV RNA as well as CMV capsid protein, with no increase in the level of accumulation of ZYMV. Resistance breakage in the resistant cultivar by ZYMV also occurred at the single cell level. Thus, synergistic interactions known to occur between a potyvirus and a cucumovirus led to resistance breakage during a double infection. However, resistance breakage was not accompanied by an increase in disease symptoms beyond those induced by ZYMV itself. On co-inoculation with an asymptomatic variant of ZYMV-AG an enhancement of CMV infection occurred without disease manifestation. Consequently, intensification of viral RNA and capsid protein accumulation can occur without a corresponding increase in disease development, suggesting that different host genes regulate viral accumulation and disease development in the CMV-resistant cucumber plants. PMID- 14745603 TI - RNA silencing as related to viroid induced symptom expression. AB - Evidence of post-transcriptional gene silencing (PTGS) in avocado infected by Avocado sunblotch viroid (ASBVd), the type species of family Avsunviroidae, was suggested by detection of ASBVd-specific 22-nucleotide RNAs. PTGS was observed in infected bleached and variegated symptomatic tissues as well as symptomless carrier foliar sources and fruit with typical sunblotch disease lesions. Tissues with the different symptom expressions, characterized by the presence of different predominant ASBVd variants, were found to induce PTGS at differential levels. Detection of the PTGS-associated small interfering RNAs (siRNAs) as well as relative concentration was also related to viroid titer. PTGS induced in Gynura aurantiaca infected with two closely-related variants of Citrus exocortis viroid, a member of family Pospiviroidae, was not directly related to viroid titer with initiation of symptoms. PMID- 14745604 TI - Variability of the progeny of a sequence variant Citrus bent leaf viroid (CBLVd). AB - A field isolate of CBLVd was previously shown to contain two dominant subpopulations (I and II), which differed by the presence or absence of a Sal I restriction site in the PCR product [10]. Here we demonstrate the infectivity and symptom expression of subpopulation II by inoculating Etrog citron with a single representative haplotype. The resulting progeny was characterised as an heterogeneous population of closely related variants with a new fitness peak represented by an haplotype that was not identified in the original isolate. This demonstrates that CBLVd conforms a "quasispecies" model. The progeny shared features of the two subpopulations of the original isolate indicating that the original isolate probably arose from a single CBLVd ancestor. PMID- 14745605 TI - Characterization and genetic diversity of potato yellow mosaic virus from the Caribbean. AB - The begomovirus Potato yellow mosaic virus (PYMV) is responsible of significant yield losses in tomato in Guadeloupe. Four field isolates from Guadeloupe were analyzed in term of their host range using three inoculation methods (mechanical, grafting and insect vector), sequences analysis of PCR fragments and phylogenetic analysis of an infectious clone, PYMV-[GP]. Capsicum annuum, Datura stramonium, Nicotiana benthamiana, N. tabacum 'Xanthi NC', Petunia hybrida, and Solanum tuberosum were found to be hosts. All isolates from Guadeloupe, Martinique, Puerto Rico and the Dominican Republic were closely related to PYMV-[GP]. Sequence identity between PYMV-[GP] and PYMV-Ve from Venezuela and PYMTV from Trinidad and Tobago clearly confirmed that it is a new strain of PYMV. PMID- 14745606 TI - Sequence analysis of both genome segments of two very virulent infectious bursal disease virus field isolates with distinct pathogenicity. AB - The deduced amino acid sequences of segment A and B of two very virulent Infectious bursal disease virus (vvIBDV) isolates, UPM94/273 and UPM97/61 were compared with 25 other IBDV strains. Twenty amino acid residues (8 in VP1, 5 in VP2, 2 in VP3, 4 in VP4, 1 in VP5) that were common to vvIBDV strains were detected. However, UPM94/273 is an exceptional vvIBDV with usual amino acid substitutions. The differences in the divergence of segment A and B indicated that the vvIBDV strains may have been derived from genetic reassortment of a single ancestral virus or both segments have different ability to undergo genetic variation due to their different functional constraints. PMID- 14745607 TI - Microscopic tumor cell dissemination in gastric cancer. AB - PURPOSE: There is still much controversy surrounding the prognostic significance of microscopic tumor cell dissemination in gastric cancer and its correlation with histopathologic parameters. We herein investigate such dissemination, predominantly restricted to the subserosa, in patients with gastric cancer. METHODS: Intraoperative bone marrow aspiration was done in 26 patients undergoing resection of gastric carcinoma. Peritoneal lavage could not be done in 8 of these 26 patients. The R0-resection rate was 84% (n = 22). A cytokeratin-directed antibody and an antibody directed against carcinoembryonic antigen served for the immunocytochemical detection of tumor cells, and the alkaline phosphatase antialkaline phosphatase method was used for visualization. RESULTS: The bone marrow aspirate and peritoneal lavage fluid were immunocytochemically positive in 31% and 56% of the patients, respectively. There was a trend (P = 0.10) towards higher overall survival rates in patients with negative bone marrow samples than in those with tumor cells detected in bone marrow samples. Analyzing the results of peritoneal lavage did not reveal any significant differences. In the group of T1/2 cancers, survival was significantly worse if the bone marrow was positive for tumor cells, with 3-year survival rates of 25% vs 77%, respectively (P < 0.05). CONCLUSION: The rates of tumor cell dissemination into the bone marrow or into the peritoneal cavity were within the scope of previous reports. Dissemination into the bone marrow resulted in significantly impaired survival in patients with T1 and T2 gastric carcinoma, and it did not correlate with known prognostic parameters. PMID- 14745608 TI - Preoperative spirometry versus expired gas analysis during exercise testing as predictors of cardiopulmonary complications after lung resection. AB - PURPOSE: As cardiopulmonary load increases with the amount of lung resected, to perform surgery safely it is important to be able to predict cardiopulmonary insufficiency. However, lung function testing with spirometry and blood gas analysis does not accurately measure cardiopulmonary reserve. We conducted this study to evaluate expired gas analysis during exercise testing for predicting postoperative complications after lung resection. METHODS: Expired gas analysis during exercise and spirometry were done 1 week preoperatively in 211 patients who underwent pulmonary resection for lung cancer. Patients were divided postoperatively according to whether cardiopulmonary complications were absent (group A) or present (group B). RESULTS: In group B there were more men than women (P < 0.01), and the mean age was greater (P < 0.05). There was no difference in disease stage, but more patients underwent pneumonectomy in group B than in group A ( P < 0.005). The results of expired gas analysis during exercise testing and of spirometry showed that maximum oxygen uptake/m(2) (P < 0.0005), anaerobic threshold/m(2) (P < 0.01), vital capacity (VC)/m(2) (P < 0.005), %VC (P < 0.0001), forced expiratory volume in 1 s (FEV(1.0))/m(2) (P < 0.0001), and FEV(1.0%) (P < 0.05) were lower in group B than in group A. CONCLUSIONS: The combination of expired gas analysis during exercise and conventional pulmonary function tests identified patients at risk for postoperative cardiopulmonary complications following pulmonary resection. PMID- 14745609 TI - Management of traumatic diaphragmatic rupture. AB - PURPOSE: Diaphragmatic rupture following trauma is often an associated and missed injury. This report documents our experience of treating traumatic diaphragmatic rupture (TDR). METHODS: We retrospectively analyzed 18 patients who presented between 1993 and 2000 with TDR, caused by blunt injuries in 14 and by penetrating injuries in 4. RESULTS: The average age of the patients was 32 years and the female to male ratio was 4 : 14. The TDR was right-sided in 5 patients and left sided in 13. The diagnosis was made by chest X-ray, thorax and upper abdominal computed tomography, and upper gastrointestinal contrast studies. The most common herniated organs were the omentum (n = 11), stomach (n = 10), spleen and colon (n = 9), and liver (n = 2). Sixteen diaphragmatic injuries were repaired primarily, and two were repaired using a prolene mesh graft. The mortality rate was 5.5% (n = 1). CONCLUSIONS: A high index of suspicion and early surgical treatment determine the successful management of TDR, with or without the herniation of abdominal organs. The surgical approach to TDR is individualized. Acute left sided injuries are best approached through the abdomen, although we prefer the chest approach, adding laparotomy when necessary. Acute right-sided injuries and chronic injuries should be approached through the chest. PMID- 14745611 TI - Stapled versus hand-sewn anastomoses in emergency intestinal surgery: results of a prospective randomized study. AB - PURPOSE: Sutured and stapled intestinal anastomoses are perceived to be equally safe in elective intestinal surgery. However, our search of the literature failed to find any studies comparing hand-sewn and mechanical anastomoses in emergency intestinal surgery. Thus, we compared the short-term outcomes of patients with sutured as opposed to stapled anastomoses in emergency intestinal surgery. METHODS: Between 1995 and 2001, 201 patients underwent emergency intestinal operations at the Department of Emergency Surgery of Sant'Orsola-Malpighi University Hospital. The outcomes of patients with sutured and stapled anastomoses were compared in a prospective analysis. Patients were randomly divided into a stapled group (106 anastomoses) with anastomoses made using linear and circular staplers, and a hand-sewn group (95 anastomoses) with anastomoses made by double-layer suturing. RESULTS: There were no significant differences between the groups in operative indications or other parameters. The operation times in the stapled group were significantly shorter than those in the hand-sewn group (P < 0.05), but there were no significant differences in anastomotic leak rates, morbidity, or postoperative mortality between the two groups. CONCLUSIONS: In emergency intestinal surgery comparable results can be achieved using mechanical and manual anastomoses. PMID- 14745610 TI - c-Met expression in a gastric cancer cell line producing alpha-fetoprotein. AB - PURPOSE: We previously reported a higher frequency of c-Met protein expression and high proliferative status in gastric cancers producing alpha-fetoprotein (AFP) than in those not producing AFP. METHODS: To investigate this further, we established an AFP-producing gastric cancer cell line, designated as AME-1, from the primary tumor of a patient with AFP-producing gastric cancer. RESULTS: alpha Fetoprotein production in the AME-1 cell line was confirmed at the protein level by immunocytochemistry and at the mRNA level by reverse transcription-polymerase chain reaction (RT-PCR). A high amount of AFP was also detected in the supernatant of cultured AME-1. The AME-1 cell line expressed c-Met both at the mRNA and protein levels. A semiquantitative RT-PCR method indicated that AME-1 had a higher amount of c-Met mRNA than well-known gastric cancer cell lines (MKN 28, MKN-45) with strong c-Met expression. Hepatocyte growth factor (HGF), a ligand for the c-Met receptor, was not detected in the mRNA or protein of AME-1. The AME-1 cell line showed a proliferative response to exogenous HGF treatment in a dose-dependent manner, but the activity of migration was not stimulated by exogenous HGF. CONCLUSIONS: The AME-1 cell line may be a useful model for elucidating aggressive behavior, especially related to regulation of the c Met/HGF system, in AFP-producing gastric cancers. PMID- 14745612 TI - Optimal strategy of preoperative transcatheter arterial chemoembolization for hepatoblastoma. AB - PURPOSE: To investigate the optimal strategy of preoperative transcatheter arterial chemoembolization (TACE) for hepatoblastoma. METHODS: Between 1992 and 2001, 7 children with hepatoblastoma (aged 9 months to 13 years) underwent preoperative TACE. The chemoembolic agent used was an emulsion of pirarubicin and lipiodol. Four patients without distant metastasis underwent "primary" TACE without systemic chemotherapy. The other 3 with distant metastases underwent "delayed" TACE following systemic chemotherapy. These patients were all examined retrospectively using clinical data. RESULTS: The average dosage of lipiodol was 0.6 ml per tumor maximal diameter (cm). All the primary cases showed a significant decrease in alpha-fetoprotein (AFP) and a reduction in the tumor size. They consequently underwent a complete surgical resection and are now disease free. All the delayed cases showed a slight decrease in AFP and underwent complete surgical resection; however, two of them died of lung metastases, and the other died of a second malignancy. Regarding complications, liver dysfunction and pulmonary embolism occurred in one patient each. CONCLUSIONS: For patients without distant metastasis, regardless of the resectability of the primary tumor, TACE may be considered the initial and only preoperative treatment, and it may be repeated. For patients with distant metastases, their complete eradication with systemic chemotherapy prior to TACE is essential. PMID- 14745613 TI - Excisional surgery for chronic advanced lymphedema. AB - PURPOSE: We conducted a retrospective study to evaluate the efficacy of excisional surgery to treat chronic advanced lymphedema. Excisional surgery was performed to reduce the extremity size, improve limb function, and decrease the frequency of lymphangitis and prevent sepsis. METHODS: Between August 1997 and December 2001, we performed excisional surgery on one or two affected legs in 20 patients with chronic lymphedema (total 24 legs). The lymphedema was graded as stage III in 5 extremities and as stage IV in 19 extremities. Five patients had primary lymphedema and 15 had secondary lymphedema. The mean duration of follow up was 17.8 months. RESULTS: Excisional surgery resulted in clinical improvement in 18 extremities, all of which received regular physiotherapy after surgery. However, lymphedema progressed to the preoperative status in six extremities, four of which did not receive regular physiotherapy after surgery. CONCLUSIONS: Excisional surgical therapy can be effective for chronic advanced lymphedema, but appropriate postoperative physiotherapy is essential to ensure that the effects last. PMID- 14745614 TI - Altered contractile responsiveness in ileal longitudinal muscle after total gastrectomy in a rat model. AB - PURPOSE: To test the hypothesis that the changes in ileal smooth muscle contractility accompanying postgastrectomy syndromes are agonist-specific, we investigated the effects of potassium chloride (KCl), carbachol, substance P (SP), and serotonin on ileal smooth muscle contractility after total gastrectomy in rats. METHODS: We performed total gastrectomy in ten rats and a sham operation in another ten rats as a control. All of the rats were killed and their ileums excised 4 weeks postoperatively. The concentration-response relationships for KCl, SP, and serotonin were obtained by adding each agent cumulatively to the organ bath. Morphological changes in the ileum were also examined by light microscopy. RESULTS: There was no significant difference between the responsiveness of gastrectomized and control tissues to KCl. Maximum responses (E(max)) to carbachol and SP were less in the gastrectomized ileal segments than in the control ileal segments. E(max) to serotonin was higher in the gastrectomized ileal segments than in the control ileal segments. The pD(2)value, i.e., the negative logarithm of the concentration for the half-maximal response, EC(50), for carbachol, SP, and serotonin was unchanged in the gastroctomized ileal segments and the control segments. Total gastrectomy also caused morphological changes in the ileum. CONCLUSIONS: These data indicate that the contractile response to various agents is altered after total gastrectomy and that receptor-mediated mechanisms may cause these changes. PMID- 14745615 TI - Teflon-felt grafting of giant gastroduodenal perforation in a canine model. AB - PURPOSE: Giant gastroduodenal perforations involving fragile and inflammatory surrounding tissue are difficult to repair with primary closure, which often results in suture tension, gastric outlet obstruction, or reperforation. However, it is still unclear if Teflon-felt can be used to effectively repair a fragile and giant gastroduodenal defect. METHODS: We made a 3.2 x 1.8-cm perforation in the gastroduodenal wall of seven adult mongrels, then grafted the defect with Teflon-felt secured with silk sutures 4 h later. Postoperatively, the dogs were fed as usual, then killed 3, 6, and 12 weeks after surgery. The results of the repair were examined carefully. RESULTS: Grafting Teflon-felt onto the giant perforations was easy to perform. Six dogs survived the operation without suffering gastric outlet obstruction, infection, or fistula formation. By the time they were killed, the dogs had lost 18%-23% of their body weight. Gross and microscopic examinations showed that the outer surface of the Teflon-felt graft was covered with omentum and fibrosis, and the undersurface was covered with slowly proliferating gastric mucosa. CONCLUSIONS: Teflon-felt appears to be a viable alternative for the delayed repair of giant ulcer perforation. PMID- 14745616 TI - Primary osteosarcoma of the lung: report of a case. AB - Osteosarcoma of the lung without an extrathoracic primary tumor is extremely rare, with only eight cases documented in the literature, to the best of our knowledge. We report a case of primary osteosarcoma of the lung found in an asymptomatic 74-year-old woman. Computed tomography showed a heterogeneous mass beside the aortic arch, and the patient underwent a left upper lobectomy. The pathology results confirmed a diagnosis of primary osteosarcoma. PMID- 14745617 TI - Extrapericardial pulmonary vein clamp technique for pulmonary arteriovenous fistula: report of a case. AB - The major concern when operating on a patient with a neurologically symptomatic pulmonary arteriovenous fistula (PAVF) is how to prevent a thromboembolic event during surgery. We describe a new technique whereby the extrapericardial pulmonary vein is clamped before transecting the afferent and efferent vessels of the fistula. The potentially pooled clots that can form while manipulating lung are stopped by the clamp. Before the extrapericardial pulmonary vein is declamped, one of the drainage veins is incised halfway and the pooled blood containing the potential clots is completely washed out. We successfully performed segmentectomy using this technique in a 66-year-old man with chronic left hemianopia and a large PAVF in the left anteromedial and lateral basal segments, and no thromboembolic events occurred. PMID- 14745619 TI - Pedunculated giant gastrointestinal stromal tumor of the stomach showing extragastric growth: report of a case. AB - We report an unusual case of a pedunculated gastrointestinal stromal tumor (GIST) of the stomach with extragastric growth. An 84-year-old woman was referred to our hospital for investigation of anemia. Barium meal examination showed a defect occupying the lesser curvature of the stomach from the middle of the corpus to the fornix and an abdominal computed tomography scan, showed a lesion, 20 cm in diameter, touching the surface of the spleen. Laparotomy revealed a tumor located between the liver and spleen. As the tumor appeared to be connected with the posterior wall of the gastric corpus by a pedicle, it was resected with a pedunculated lesion of the stomach. Based on the pathological findings, the tumor was diagnosed as a GIST of uncommitted type, with low-grade malignancy. PMID- 14745618 TI - Simultaneous esophagectomy and coronary artery bypass grafting without cardiopulmonary bypass: report of a case. AB - We successfully performed off-pump coronary artery bypass grafting (OPCAB) with concomitant esophagectomy in a 77-year-old man with esophageal cancer and severe stenosis of the anterior descending branch of the left coronary artery. Off-pump coronary artery bypass grafting was performed via median sternotomy and esophagectomy was done via the left thoracoabdominal approach. The patient was discharged with a patent graft 8 weeks after surgery. The benefits of OPCAB include that it is less invasive and heparinization can be avoided. This case report demonstrates that simultaneous OPCAB and esophagectomy is advantageous for a selected population with surgically correctable coronary artery disease and resectable esophageal cancer. PMID- 14745620 TI - Multiple jejunal diverticulitis with perforation in a patient with systemic lupus erythematosus: report of a case. AB - A 70-year-old man with systemic lupus erythematosus (SLE) was brought to our Emergency Department after the sudden onset of acute and severe abdominal pain. Physical examination revealed a tender and distended abdomen with guarding and rebound tenderness in the periumbilical region and the left upper quadrant. A plain abdominal X-ray taken with the patient upright showed air fluid levels with dilatation of several loops in the small bowel. As the examination could not rule out bowel ischemia, perforation, or obstruction, an emergency laparotomy was performed, which revealed multiple jejunal diverticulosis, one of which had perforated and adhered to the right colon, causing rotation. The diverticulosis segment was resected and an end-to-end anastomosis was done. The patient had an uneventful postoperative recovery without any complications. This is an unusual cause of peritonitis in a patient with SLE, and we could not find any evidence to suggest involvement of the underlying SLE in the jejunal diverticulosis and diverticulitis in this patient. Nevertheless, the involvement of SLE might be possible and further investigation is warranted. PMID- 14745621 TI - Heterotopic ossification of rectal adenocarcinoma: report of a case. AB - Heterotopic ossification of gastrointestinal tract tumors is rare. We report the case of a 67-year-old man with heterotopic ossification of a rectal adenocarcinoma. The patient presented with intermittent abdominal pain and frequent diarrhea, and colonoscopic examination showed a large polypoid tumor partially obstructing the rectal lumen. Abdominal computed tomography (CT) revealed a large tumor in the rectal lumen with calcified spots. We performed low anterior resection of the rectum, and histologic examination showed a well differentiated adenocarcinoma with heterotopic ossification infiltrating the full thickness of the rectum. Local recurrence and liver metastases were found 2 months after surgery, and the patient died 3 months later. Such a rapidly progressive course of rectal adenocarcinoma with heterotopic ossification is very unusual. PMID- 14745622 TI - Anorectal agenesis with a rectourethral fistula diagnosed in an adult: report of a case. AB - We report an unusual case of anorectal agenesis with a rectourethral fistula diagnosed in a 48-year-old man. The patient presented after noticing hematuria, although he had been aware of urinary leakage from his colostomy with occasional fecal urine for about 4 years. He had had a double-barrel colostomy created soon after birth for an imperforate anus, with revision at the age of 4 years to correct a prolapse of the stoma, but his malformation had never been repaired. We performed a physical examination, which did not reveal a perineal fistula, but urethrocystography demonstrated high anorectal agenesis with a rectourethral fistula. Thus, we resected the rectourethral fistula and created an end colostomy. The patient had an uneventful postoperative course, and was discharged in good health on postoperative day 19. To our knowledge, this is the oldest patient to be diagnosed with anorectal agenesis and undergo resection of a rectourethral fistula. PMID- 14745623 TI - Septic thrombophlebitis of the portal and superior mesenteric veins as a complication of appendicitis: report of a case. AB - Pylephlebitis is extremely rare and associated with high mortality, even in this modern era. It usually occurs secondary to infection in the region drained by the portal systems or in the structure contiguous to the portal vein. We report a case of septic thrombophlebitis of the portal and superior mesenteric veins (SMV) with multiple liver abscesses caused by acute appendicitis with an abscess of the mesoappendix. We performed appendectomy and successfully removed the thrombi using a Fogarty catheter. Postoperative histopathological examination confirmed a diagnosis of appendicitis and septic thrombophlebitis of the portal vein and SMV. The patient recovered completely with appropriate medical and surgical treatment. PMID- 14745624 TI - Small nonfunctioning islet cell tumor in the body of the pancreas: report of a case. AB - Islet cell tumors of the pancreas are uncommon, and nonfunctioning tumors are even rarer than functioning ones. We report the case of a 67-year-old woman with a small nonfunctioning islet cell tumor, 6 x 5 mm in diameter, which was detected incidentally by ultrasonography, and subsequently confirmed by double-helical computed tomography. Diagnosis was established by histopathological examination after 80% distal pancreatectomy with splenectomy, and by various laboratory tests. Histologically, the islet cell tumor showed highly cellular spindle or epithelioid cells, which were positive for Grimelius stain. Immunohistochemical examination revealed that the tumor cells were positive for chromogranin A, but negative for somatostatin, insulin, glucagon, and gastrin. Its small size, location, and benignity make this a very rare type of nonfunctioning islet cell tumor. PMID- 14745625 TI - Pseudoaneurysm of the superior pancreaticoduodenal artery, a rare cause of hemosuccus pancreaticus: report of a case. AB - Chronic pancreatitis with a pseudoaneurysm is an established cause of hemosuccus pancreaticus. We herein describe a patient with chronic alcoholic pancreatitis associated with hemosuccus pancreaticus due to a pseudoaneurysm of the anterior superior pancreaticoduodenal artery rupturing in a pseudocyst of pancreas in the head region. Angiographic embolization was unsuccessful and therefore a laparotomy, ligation, and excision of the pseudoaneurysm with external drainage of pseudocyst were performed. Hemosuccus pancreaticus is a rare cause of upper gastrointestinal bleeding. Contrast-enhanced computed tomography and angiography is diagnostic in the majority of the cases. Surgery is the treatment of choice. Selective angiographic embolization may be helpful in tiding over the emergency until surgery can be performed. PMID- 14745626 TI - Aortobifemoral graft causing complete intestinal obstruction in a patient with Behcet's disease: report of a case. AB - A 50-year-old man with Behcet's disease and a history of aortobifemoral (ABF) bypass grafting for an abdominal aortic aneurysm was admitted to our Emergency Room with ileus. Upon exploration we found that the ABF graft was occluding the jejunal passage. To the best of our knowledge this is the first such case ever to be reported. PMID- 14745627 TI - Surgical treatment for an atherosclerotic aneurysm of the superficial femoral artery: report of a case. AB - We report the case of an isolated atherosclerotic aneurysm of the superficial femoral artery (SFA). The patient was a 76-year-old man who presented with intermittent claudication and muscle weakness in his right leg. Angiography showed an aneurysm in the lower portion of the right SFA, but no abdominal aortic aneurysm or any other peripheral aneurysms were detected. The patient had not suffered any major complications such as rupture, thrombosis, or distal emboli by the time of presentation. Thus, we resected the aneurysm and replaced it with a Dacron graft. Pathological examination revealed typical atherosclerotic changes and no evidence of infection, previous injury, or connective tissue disorders. While isolated atherosclerotic aneurysms of the SFA are rare, with only about 30 cases documented, patients frequently present with major complications and therefore, making an early diagnosis and initiating appropriate surgical treatment are essential. PMID- 14745628 TI - Incidence of anterior displacement of the anus and its relationship to constipation in children. AB - Anterior displacement of the anus (ADA), otherwise known as "anterior ectopic anus," is considered to be a common congenital abnormality of anorectal region. However, the true incidence of this condition is not well known. We evaluated the incidence of ADA and its relationship to constipation in 357 children (191 boys and 166 girls) who were admitted to our hospital with unrelated disorders. For this purpose, the anal index was defined as the ratio of the scroto-anal distance to the scroto-coccygeal distance in boys, and as the ratio of the fourchette-anal distance to the fourchette-coccygeal distance in girls. An anal index of less than 0.34 in girls and less than 0.46 in boys was determined to be an ADA. The incidence of ADA was 43.4% in girls and 24.6% in boys (P < 0.01). However, the incidence of constipation in children with a normal anal index and those with a low anal index indicative of an ADA was not significantly different. These findings suggest that ADA is a common variant of the normal anatomical location of the anus, especially in girls. PMID- 14745629 TI - External tissue expansion successfully achieved using negative pressure. AB - The practice of internal tissue expansion by implanting a silicone balloon into the soft tissue under the skin is now known to be associated with a significant risk of complications. Therefore, we designed and developed a new technique of mechanical external tissue expansion achieved by using negative pressure created through special valved cups. The subjects were 60 patients suffering from postburn scar contractures in different parts of their bodies. A valved cup was applied to the healthy skin on both sides of the scar and negative pressure of 100 to -200 mbar was connected to the cup through a special valve for 5 h out of every 12 h for 2-3 weeks preoperatively. During the subsequent operation, the scar was excised, and then the expanded skin was dissected followed by wound closure. We were able to excise the scars and close the wounds directly in 14 patients, but needed to perform Z-plasty in the other 46 patients. Some ecchymosis appeared on the skin at the site of application of the cup in the initial stages, when the negative pressure exceeded the upper limits of -100 to 200 mbar. The clinical application of external tissue expansion achieved by creating negative pressure shows great promise. PMID- 14745630 TI - Boron uptake by ectomycorrhizas of silver birch. AB - Boron (B) is an essential micronutrient for plants but it is thought not to be essential for fungi. We studied whether the extraradical mycelia of Paxillus involutus in symbiosis with silver birch (Betula pendula) take up B and transport it to the host plant. We grew mycorrhizal plants in flat microcosms with a partitioning wall, below which there was only extraradical mycelium. A boric acid solution enriched in 10B was applied to these mycelia. Increased 10B/11B isotope ratios were subsequently measured in birch leaves, stems, and roots plus mycorrhizas in the upper compartment. Boron was therefore taken up by the mycorrhizal mycelia and transported to the host plant in this species combination. PMID- 14745631 TI - Morphological and molecular characterization of selected Ramaria mycorrhizae. AB - Ramaria species are conspicuous mycorrhizal symbionts of conifers in the Pacific Northwest. Here we collected and identified sporocarps and associated ectomycorrhizae of Ramaria acrisiccescens Marr & Stuntz, R. cyaneigranosa Marr & Stuntz, R. sandaracina Marr & Stuntz, R. celerivirescens Marr & Stuntz, and R. flavobrunnescens var. aromatica Marr & Stuntz. An internal transcribed spacer (ITS)- restriction fragment length polymorphism pattern was observed for each of the Ramaria species and used as a diagnostic tool to support the identification of mycorrhizae occurring in mats below the sporocarps. We provide a description of ectomycorrhizae of Ramaria, which exhibit similar macro- and microscopic characteristics such as ramification pattern, coloration, abundance of mycelial strands and emanating hyphae, mantle morphology and chemical reactions of mantle and mycelial strands with KOH, FeSO4 and Melzer's reagent. Sequences of the ITS region for each of the species are deposited in the GenBank. PMID- 14745632 TI - Acute renal failure associated with Gemella haemolysans pneumonia. AB - We describe a 13-year-old boy who developed acute renal failure associated with Gemella haemolysans pneumonia. At presentation he was found to have macroscopic hematuria associated with lobar pneumonia. Gemella haemolysans was isolated from blood cultures. Renal failure was detected on admission, progressed and dialysis was required until day 18. Renal impairment had resolved by 3 months after the initial presentation. Histopathology of a renal biopsy showed focal proliferative glomerulonephritis, but the predominant abnormality was tubular damage associated with erythrocyte casts in tubular lumina. We believe that tubular damage due to hematuria rather than the glomerular changes was the most likely cause of renal failure. PMID- 14745633 TI - Soluble adhesion molecules in children and young adults on chronic hemodialysis. AB - Children on chronic hemodialysis (HD) present with impaired immunity that may result from disturbances in leukocyte migration, caused by changes in expression of adhesion molecules on endothelium and immunocompetent cells. However, it is still not clear whether the type of dialyzer or a single dialysis session influences the concentrations of soluble adhesion molecules in these patients. We evaluated by ELISA serum levels of soluble (s) VCAM-1, ICAM-1, L-selectin, and P selectin in 22 patients on cuprophane HD (CU), 8 on polysulfone HD (PS), 10 on vitamin E-modified cellulose HD (VE), and 15 controls. In all HD patients, sVCAM 1 levels were elevated compared with controls and were higher in CU than in VE. The sICAM-1 concentrations were increased in VE compared with controls, but remained unchanged in CU and PS. The sL-selectin levels were reduced in all HD patients. The mean values of sP-selectin were comparable in CU, PS, and controls. The lowest levels were observed in VE. In CU patients, sVCAM-1, sICAM-1, and sP selectin concentrations rose after HD. A single PS session had no impact on adhesion molecules, whereas a VE session increased the level of sVCAM-1. The type of dialysis membrane may change the profile of adhesion molecule concentrations, thus influencing the immune system of a child on HD. The increase in levels of adhesion molecules in the course of a single HD session, which was pronounced in CU and VE patients, suggests poor biocompatibility of these dialyzers. PMID- 14745634 TI - Clinical experience with darbepoietin alfa (NESP) in children undergoing hemodialysis. AB - Darbepoietin alfa (NESP) is a new long-acting erythropoietin, with a half-life 3 times longer than the old epoietins. In the present study, we evaluated the efficacy of NESP in a group of children on hemodialysis. Seven children, five male and two female, with a mean age of 11.5 +/- 3 years and a mean weight of 34.1 +/- 11 kg, were enrolled in the study. All had been treated for at least 6 months with epoietin alfa at a mean dose of 106 +/- 76 IU/kg 3 times/week i.v. They were then given NESP at a mean dose of 1.59 +/- 1.19 microg/kg once a week i.v., according to the suggested conversion index (weekly epoietin alfa dose/200=weekly NESP dose). Anemia was evaluated at the end of a dialysis session. This was especially important for children less compliant with water restriction. Serum ferritin and percentage transferrin saturation (TSAT) were also monitored, as were dialysis efficacy (Kt/V), blood pressure, and heparin requirements. Before starting the new treatment, all patients had an adequate mean hemoglobin (Hb) level (11.19 +/- 1.7 g/dl) and an adequate iron status (TSAT 24.2 +/- 11.5, serum ferritin 220 +/- 105 mg/dl). Five of the seven patients were also treated with intravenous ferric gluconate (10-20 mg/kg per week). Six children were on antihypertensive treatment. After the 1st month of treatment, we observed an excessive increase in Hb, 12.3 +/- 1.7 g/dl, (P<0.05), with severe hypertension in the youngest two patients (Hb>13 g/dl). A short discontinuation of the medication, followed by restarting at a decreased dosage, allowed us to continue with the treatment. At the 2nd month of follow-up, a mean plasma Hb level of 12.2 +/- 1.2 g/dl was observed, with a NESP mean dose of 0.79 +/- 0.4 microg/kg per week. Steady state was reached at 3 months, with a mean Hb of 11.8 +/- 1.4 g/dl and a mean NESP dose of 0.51 +/- 0.18 microg/kg per week (P<0.05). These results persisted at 6 months of follow-up; only one child had a persistent increase in platelet level (373,000 vs. 555,000). Dialysis efficiency and heparin requirements during dialysis did not change significantly. The high efficacy of NESP allowed a consistent reduction in dosage. The suggested conversion index does not appear to be correct for pediatric patients. Our experience suggests that in this population the correct dose could be 0.25-0.75 microg/kg per week. Hypertension was the only major side effect reported. The influence of NESP on platelet proliferation needs to be further investigated. The single weekly administration of NESP could be effective and beneficial for both patients and clinicians. PMID- 14745636 TI - Slow progressive FSGS associated with an F392L WT1 mutation. AB - Constitutional missense mutations in the WT1 gene are usually associated with the Denys-Drash syndrome, characterized by a rapid progressive nephropathy, male pseudohermaphroditism, and an increased risk for Wilms tumor. We report here a patient with scrotal hypospadias and a slow progressive nephropathy due to focal and segmental glomerulosclerosis. WT1 mutation analysis revealed a constitutional missense mutation in exon 9 resulting in an exchange F392L. This mutation has previously been reported by others in a patient with a similar mild course of nephropathy. In contrast, a mutation in the corresponding codon of exon 8 (F364L) was previously found by us in a patient with a very rapid progression to end stage renal disease. Whether the position of a mutation may influence the course of the nephropathy must be evaluated in a larger patient cohort. The individual tumor risk for this alteration cannot be given at present because neither of the two patients has shown evidence of a Wilms tumor or a gonadoblastoma to date. PMID- 14745635 TI - Enalapril in children with Alport syndrome. AB - Ten pediatric patients with Alport syndrome received enalapril for 5 years. There were nine boys. Eight patients have the X-linked form of the disease and two the autosomal recessive form. The median age at the start of treatment was 10.25 years. Only one patient was hypertensive. The starting dose of enalapril was 0.05 mg/kg; the target dose was 0.5 mg/kg per day. The median dose given effectively was 0.24, 0.37, 0.45, 0.43, and 0.49 mg/kg per day at years of study 1, 2, 3, 4, and 5, respectively. The median urinary protein/creatinine ratio was 1.58 g/g (range 0.49-4.60) before treatment. This decreased to 0.98, 1.09, 1.35, 1.11, and 1.38 g/g after 1, 2, 3, 4, and 5 years, respectively. The median creatinine clearance at baseline was 100 ml/min per 1.73 m2 (range 82-133) and after 5 years 92 ml/min per 1.73 m2 (range 22-115). Three patients did not reach the target dose of enalapril because of orthostatic hypotension. One of them was the only patient to develop chronic renal failure within 5 years. The present study indicates that enalapril reduces urinary protein excretion and preserves glomerular filtration in Alport patients as a group. However, there was individual variation, as in most studies of patients with proteinuric nephropathies given inhibitors of the angiotensin-converting enzyme. PMID- 14745637 TI - Hyponatremia and hyperkalemia in infants with acute pyelonephritis. PMID- 14745638 TI - Sterile pyuria in Kawasaki disease. PMID- 14745639 TI - Modulation of voluntary swallowing by visual inputs in humans. AB - The purpose of this study was to test the hypothesis that a stimulus which strengthens a central input to a swallowing-related cortical area, given before voluntary swallowing, could facilitate subsequent swallowing movements. The subjects consisted of seven healthy volunteers. We used visual images to strengthen central input. The subjects voluntarily performed either dry swallowing or water swallowing after presentation of the visual images. Under the water-swallowing condition, the latency was significantly shorter and the maximum amplitude of the suprahyoid electromyographic (EMG) activity was significantly smaller in subjects who received drink-related visual input. However, there were no similar differences under the dry-swallowing condition. In addition, there were no significant differences in the mean EMG amplitude or the duration of EMG activity between subjects who did and did not receive drink-related visual input under either swallowing condition. We concluded that drink-related visual inputs prior to voluntary swallowing facilitate the initiation of swallowing and enhance swallowing-related muscle activity in the presence of peripheral inputs. PMID- 14745640 TI - Characteristics of dysphagia in elderly patients requiring mechanical ventilation. AB - The purpose of this study was to describe the swallowing characteristics of elderly patients requiring mechanical ventilation with tracheostomy admitted to a long-term, acute-care hospital. The study was conducted through retrospective record review of patients on mechanical ventilation who had received a Modified Barium Swallow Study (MBSS) during their hospitalization. In a period from 1994 to 2002, 58 patients met the inclusion criteria. The study examined the results of both the clinical and the MBSS evaluations and compared the results and recommendations of the two examinations. Data were obtained from the MBSS records to describe the group in terms of dysphagia symptoms, frequency and occurrence of aspiration, respiratory status, and demographic variables. Parametric and nonparametric statistics were used to determine differences between the evaluations and any significant associations between aspiration and demographic variables, pharyngeal symptoms, and cognitive deficits. Significant differences were found between diet recommendations before and after the MBSS, and significant associations were found between aspiration and three pharyngeal symptoms. Although aspiration and especially silent aspiration occurred frequently in this group, most individuals were able to begin some level of oral intake after the MBSS evaluation. Due to the lack of reliable clinical evaluation measures, the MBSS is necessary for differential diagnosis of dysphagia and dietary recommendations for these individuals. PMID- 14745641 TI - Assessment of the predictors of response to glucagon in the setting of acute esophageal food bolus impaction. AB - Esophageal food impactions are frequently seen in endoscopic practice. Glucagon is known to relax the lower esophageal sphincter and has been used with variable success to treat food impactions. We retrieved clinical information of all patients with acute food impactions who attended the emergency room from 1975 to 2000 from the Mayo diagnostic database. Data were abstracted on age, sex, body mass index, relevant prior medical history, food type ingested (meat, bread, vegetable, or other), duration of symptoms at presentation, dosage (in mg) of glucagon, outcome including success of glucagon or spontaneous passage, and endoscopic findings. A total of 222 cases of food impaction were identified, of whom 106 patients (48%) received glucagon, average 1 mg. In glucagon responders, meat was less likely to be the offending food type, accounting for 70% (glucagon responders) vs. 90% (in nonresponders) ( p = 0.03), while responders were less likely to have esophageal rings/strictures detected on subsequent EGD compared with nonresponders, 0% (glucagon responders) vs. 31% (nonresponders) ( p = 0.05). In the patients that did not receive glucagon, spontaneous resolvers had a shorter duration of symptoms at presentation, 3.3 h vs. 12.4 h ( p = 0.07) and were less likely to have an organic esophageal obstruction detected on EGD, 0% vs. 21%. There were no significant differences between the resolvers and nonresolvers in terms of age, gender, BMI, and prior medical history. Conservative management of acute food bolus obstruction, either with or without glucagon, is most successful in the absence of a fixed esophageal obstruction. An impacted meat bolus is more likely to require intervention for removal than other food types. These clinical predictors should be considered before administration of glucagon. PMID- 14745642 TI - Endoscopic intubation with conventional plastic stents: a safe and cost-effective palliation for inoperable esophageal cancer. AB - Access to expensive equipment and costly self-expanding metal endoprostheses is limited in some regions where unresectable esophageal cancer is not infrequent. The aim of this study was to review the long-term results of palliation of malignant esophageal obstruction using low-priced conventional plastic stents. One hundred sixty-nine patients with dysphagia due to inoperable esophageal cancer underwent esophageal intubation under endoscopic control alone, without general anesthesia, by the pulsion method. Stents mounted on their delivery device were inserted over an endoscopically placed guide wire. Improvement in swallowing was seen in all patients. Dysphagia scores have improved from 3.64 +/- 0.21 to 1.08 +/- 0.17. Major early procedure-related morbidity was high at 0.6% with one intramural perforation (no transmural perforation at all). Minimal mucosal bleeding was seen with 72 cases (42.6%). Procedure-related mortality was 0%. Late procedure-related complications requiring further endoscopic procedures occurred in 8.2% (tube occlusion: 5.3%, tube dislocation: 2.9%). Our 7-day mortality was 0% and 5 patients died within 30 days, usually from the disease itself. Those surviving the procedure (more than 7 days) had a mean survival of 209 days. Esophageal plastic stents can be accurately and safely placed under direct endoscopic control with lower costs. Therefore, endoscopic intubation remains a useful palliative treatment for patients with unresectable carcinoma of the esophagus. PMID- 14745643 TI - Awareness of dysphagia by patients following stroke predicts swallowing performance. AB - Patients' awareness of their disability after stroke represents an important aspect of functional recovery. Our study aimed to assess whether patient awareness of the clinical indicators of dysphagia, used routinely in clinical assessment, related to an appreciation of "a swallowing problem" and how this awareness influenced swallowing performance and outcome in dysphagic stroke patients. Seventy patients were studied 72 h post hemispheric stroke. Patients were screened for dysphagia by clinical assessment, followed by a timed water swallow test to examine swallowing performance. Patient awareness of dysphagia and its significance were determined by detailed question-based assessment. Medical records were examined at three months. Dysphagia was identified in 27 patients, 16 of whom had poor awareness of their dysphagic symptoms. Dysphagic patients with poor awareness drank water more quickly (5 ml/s vs. <1 ml/s, p = 0.03) and took larger volumes per swallow (10 ml vs. 6 ml, p = 0.04) than patients with good awareness. By comparison, neither patients with good awareness or poor awareness perceived they had a swallowing problem. Patients with poor awareness experienced numerically more complications at three months. Stroke patients with good awareness of the clinical indicators of dysphagia modify the way they drink by taking smaller volumes per swallow and drink more slowly than those with poor awareness. Dysphagic stroke patients, regardless of good or poor awareness of the clinical indicators of dysphagia, rarely perceive they have a swallowing problem. These findings may have implications for longer-term outcome, patient compliance, and treatment of dysphagia after stroke. PMID- 14745644 TI - Oropharyngeal scintigraphy: a reliable technique for the quantitative evaluation of oral-pharyngeal swallowing. AB - A valid and reliable technique to quantify the efficiency of the oral-pharyngeal phase of swallowing is needed to measure objectively the severity of dysphagia and longitudinal changes in swallowing in response to intervention. The objective of this study was to develop and validate a scintigraphic technique to quantify the efficiency of bolus clearance during the oral-pharyngeal swallow and assess its diagnostic accuracy. To accomplish this, postswallow oral and pharyngeal counts of residual for technetium-labeled 5- and 10-ml water boluses and regional transit times were measured in 3 separate healthy control groups and in a group of patients with proven oral-pharyngeal dysphagia. Repeat measures were obtained in one group of aged (> 55yr) controls to establish test-retest reliability. Scintigraphic transit measures were validated by comparison with radiographic temporal measures. Scintigraphic measures in those with proven dysphagia were compared with radiographic classification of oral vs. pharyngeal dysfunction to establish their diagnostic accuracy. We found that oral ( p = 0.04), but not pharyngeal, isotope clearance is swallowed bolus-dependently. Scintigraphic transit times do not differ from times derived radiographically. All scintigraphic measures have extremely good test-retest reliability. The mean difference between test and retest for oral residual was -1% (95% CI -3%-1%) and for pharyngeal residual it was -2% (95% CI -5%-1%). Scintigraphic transit times have very poor diagnostic accuracy for regional dysfunction. Abnormal oral and pharyngeal residuals have positive predictive values of 100% and 92%, respectively, for regional dysfunction. We conclude that oral-pharyngeal scintigraphic clearance is highly reliable, bolus volume-dependent, and has a high predictive value for regional dysfunction. It may prove useful in assessment of dysphagia severity and longitudinal change. PMID- 14745645 TI - Evaluating swallowing dysfunction using a 100-ml water swallowing test. AB - This study used comparison with videofluoroscopic examination of swallowing (VFES) to examine the validity of a 100-ml water swallowing test (WST) in assessing swallowing dysfunction. Fifty-nine consecutive outpatients (15 females, 44 males) with clinically suspected dysphagia were enrolled in this study. Each subject underwent a 100-ml WST followed by VFES. Data was obtained on swallowing speed and signs of choking (coughing and a wet-hoarse voice). The analytical results revealed that 49 subjects had abnormal swallowing speeds (< 10 ml/s) in the 100-ml WST, and 47 of them were identified as having dysphagia by VFES. Among the ten participants with normal swallowing speed (> 10 ml/s), eight were diagnosed with dysphagia by VFES. Notably, 14 participants choked in the 100-ml WST, 11 of whom exhibited aspiration or penetration in VFES. Among the 45 participants without choking in WST, 12 displayed aspiration or penetration in VFES. The sensitivity of swallowing speed in detecting the swallowing dysfunction was 85.5%, and the specificity was 50%. Moreover, the sensitivity of using choking or wet-horse voice in the 100-ml WST as the sole factor for predicting the presence of aspiration was 47.8%, while the specificity was 91.7%. Therefore, this study concluded that swallowing speed is a sensitive indicator for identifying patients at risk for swallowing dysfunction. Moreover, choking in the 100-ml WST may be a potential specific indicator for followup aspiration. PMID- 14745646 TI - Postcricoid hemangioma: an overlooked cause of dysphagia in infants?-a case report. AB - Feeding and swallowing disorders in children remain a major challenge owing to a wide differential diagnosis. Hemangioma of the upper aerodigestive tract represents one of the numerous non-neoplastic causes of dysphagia. We report two cases of postcricoid hemangioma causing inhalation and recurrent respiratory infections, treated successfully with systemic corticotherapy alone. To our knowledge, these are the second and third cases described in the literature. After a short review of the literature, the diagnostic procedures are discussed and a management strategy is proposed for this clinical entity, by far underestimated. PMID- 14745647 TI - Prolonged effect of botulinum toxin injection in the treatment of cricopharyngeal dysphagia: case report and literature review. AB - Cricopharyngeus (CP) muscle spasm can lead to severe dysphagia. Myotomy of the CP muscle was the treatment of choice. Recently, botulinum toxin type A (BtxA) has been used for CP spasm. It usually brings improvement in deglutition but most patients require reinjection in 3-5 months. We report a 35-year-old man who had an arteriovenous malformation hemorrhage in the brain stem resulting in CP spasm and consequently severe dysphagia. He received BtxA injection and deglutition and nutrition remained good one year after treatment. A literature review analyzing 28 patients and our patient showed negative correlations between age and BtxA dose and between age and duration. Efficacy was positively correlated with duration and BtxA dose was positively correlated with pretreatment severity. In conclusion, physicians would use higher doses on patients with more severe cases but use lower doses on older patients. Those who obtained better post-treatment results would enjoy longer effective duration. Thus, the effective duration of the BtxA is multifactorial. PMID- 14745648 TI - Allometric constraints on Sr/Ca and Ba/Ca partitioning in terrestrial mammalian trophic chains. AB - In biological systems, strontium (Sr) and barium (Ba) are two non-essential elements, in comparison to calcium (Ca) which is essential. The Sr/Ca and Ba/Ca ratios tend to decrease in biochemical pathways which include Ca as an essential element, and these processes are termed biopurification of Ca. The quantitative pathway of the biopurification of Ca in relation to Sr and Ba between two biological reservoirs ( Rn and R(n -1)) is measured with an observed ratio (OR) expressed by the (Sr/Ca) Rn /(Sr/Ca)( Rn-1) and (Ba/Ca) Rn /(Ba/Ca)( Rn-1) ratios. For a mammalian organism, during the whole biopurification of Ca starting with the diet to the ultimate reservoir of Ca which is the bone, the mean values for ORSr and ORBa are 0.25 and 0.2, respectively. In this study, published Sr/Ca and Ba/Ca ratios are used for three sets of soils, plants, and bones of herbivorous and carnivorous mammals, each comprising a trophic chain, to illustrate the biopurification of Ca at the level of trophic chains. Calculated ORSr and ORBa of herbivore bones in relation to plants and of bones of carnivores in relation to bones of herbivores give ORSr=0.30+/-0.08 and ORBa=0.16+/-0.08, thus suggesting that trophic chains reflect the Sr/Ca and Ba/Ca fluxes that are prevalent at the level of a mammalian organism. The slopes of the three regression equations of log(Sr/Ca) vs. log(Ba/Ca) are similar, indicating that the process of biopurification of Ca with respect to Sr and Ba is due to biological processes and is independent of the geological settings. Modifications of the logarithmic expression of the Sr/Ca and Ba/Ca relationship allow a new formula of the biopurification process to be deduced, leading to the general equation ORBa=ORSr(1.79+/-0.33), where the allometric coefficient is the mean of the slopes of the three regression equations. Some recent examples are used to illustrate this new analysis of predator-prey relations between mammals. This opens up new possibilities for the utilization of Ba/Ca and Sr/Ca in addition to stable isotope ratios (delta13C and delta15N) for the determination of the relative contribution of different food sources to an animal's diet. PMID- 14745649 TI - Fruiting trees as dispersal foci in a semi-deciduous tropical forest. AB - Quantification of seed rain patterns is an initial step toward explaining variation in plant recruitment, and consequently, organization of forest communities. Spatially contagious patterns of seed deposition, where seeds are patchily dispersed with some sites receiving relatively high densities and others receiving low densities of seeds, may be a common phenomenon for which we have very little knowledge. For example, prior feeding events by frugivores (monkeys and birds) combined with transport and dispersal of seeds to other fruiting trees may result in the contagious deposition of non-conspecific seeds below them. Here, we examined whether fruiting trees act as dispersal foci in the semi deciduous tropical rainforest of the Dja Reserve, Cameroon. Seed rain was sampled below the canopies of nine tree species: three typically dispersed by large, frugivorous birds, three dispersed by monkeys, and three dispersed by wind. We found no evidence that monkeys generate spatially contagious patterns of seed rain under fruiting trees at which they feed. However, we found that rates of deposition of non-conspecific seeds and species richness of seeds delivered by birds (hornbills and turacos) were significantly greater during fruiting than non fruiting periods, and significantly greater under fruiting individuals of bird dispersed tree species than under fruiting individuals of monkey- or wind dispersed tree species. Additionally, during fruiting periods, the composition of non-conspecific seed rain under bird-dispersed tree species was more similar to other bird-dispersed trees than to monkey- or wind-dispersed tree species. The contagious dispersal of non-conspecific seeds to fruiting, bird-dispersed trees leads to higher seed densities under fruiting trees than those caused by local seed production. Non-conspecific seeds deposited in high densities may experience increased seed mortality even far from parent trees if predators are generalists. Alternatively, in the absence of complete density-dependent mortality, contagious seed dispersal could result in associations among species dispersed by the same dispersal agent. PMID- 14745650 TI - Direct and ecological costs of resistance and tolerance in the stinging nettle. AB - Plant resistance and tolerance to herbivores, parasites, pathogens, and abiotic factors may involve two types of costs. First, resistance and tolerance may be costly in terms of plant fitness. Second, resistance and tolerance to multiple enemies may involve ecological trade-offs. Our study species, the stinging nettle ( Urtica dioicaL.) has significant variation among seed families in resistance and tolerance as well as costs of resistance and tolerance to the holoparasitic plant Cuscuta europaea L. Here we report on variation among seed families (i.e. genetic) in tolerance to nutrient limitation and in resistance to both mammalian herbivores (i.e. number of stinging trichomes) and an invertebrate herbivore (i.e. inverse of the performance of a generalist snail, Arianta arbustorum). Our results indicate direct fitness costs of snail resistance in terms of host reproduction whereas we did not detect fitness costs of mammalian resistance or tolerance to nutrient limitation. We further tested for ecological trade-offs among tolerance or resistance to the parasitic plant, herbivore resistance, and tolerance to nutrient limitation in the stinging nettle. Tolerance of nettles to nutrient limitation and resistance to mammalian herbivores tended to correlate negatively. However, there were no significant correlations among resistance and tolerance to the different natural enemies (i.e. parasitic plants, snails, and mammals). The results of this greenhouse study thus suggest that resistance and tolerance of nettles to diverse enemies are free to evolve independently of each other but not completely without direct costs in terms of plant fitness. PMID- 14745652 TI - Management of amiodarone-induced thyrotoxicosis. AB - Amiodarone is used increasingly in a number of cardiac conditions. Amiodarone is heavily iodinated and can cause thyroid dysfunction. The diagnosis of amiodarone induced thyrotoxicosis remains difficult and more common causes of thyrotoxicosis need to be considered and excluded. Amiodarone has a significant side effect profile, which includes thyroid dysfunction. Amiodarone is an effective drug and its withdrawal may have significant impact on a patient's already fragile cardiac status. There are three different types of amiodarone-induced thyrotoxicosis (AIT) (I, II and mixed). Identification of the different subtypes of AIT allows a rational and appropriate management strategy to be chosen. Type I occurs in patients with underlying thyroid disease, whilst type II is thought to result from a destructive thyroiditis. Differentiation is based on clinical grounds together with investigations, which can include thyroid function test, radioiodine uptake scanning, measurement of interleukin-6 levels and colour flow Doppler sonography. Amiodarone should be discontinued in both types of AIT if the indication for its use is not a life-threatening cardiac condition. The management of type I centres around antithyroid drugs to control thyrotoxicosis and later consideration of more definitive treatment. Type II AIT responds to steroid therapy, although antithyroid drugs may be useful if symptoms are severe. Therapeutic options for refractory cases of AIT include surgery, radioiodine and plasmapheresis. PMID- 14745651 TI - Common variants within the interleukin 4 receptor alpha gene (IL4R) are associated with susceptibility to osteoarthritis. AB - Primary osteoarthritis (OA) is a common late-onset arthritis that demonstrates a complex mode of transmittance with both joint-site and gender-specific heterogeneity. We have previously linkage-mapped an OA susceptibility locus to a 12-cM interval at chromosome 16p12.3-p12.1 in a cohort of 146 affected female sibling-pair families ascertained by total hip replacement (female-THR families), with a maximum multipoint LOD score of 1.7. Despite the low LOD score, we were encouraged to investigate this interval further following the report of a linkage to the same interval in an Icelandic pedigree with an early-onset form of hip OA. Using public databases, we searched the interval for plausible candidates and concluded that the gene encoding the interleukin 4 receptor alpha chain (IL4R) was a particularly strong candidate based on its known role in cartilage homeostasis. We genotyped nine common single nucleotide polymorphisms (SNPs) from within IL4R, including six non-synonymous SNPs, in the 146 probands from our female-THR families (stage 1) and in an independent cohort of 310 female-THR cases (stage 2). We compared allele frequencies with those of 399 age-matched female controls. All individuals were UK Caucasians. The minor alleles of two SNPs demonstrated association in both stages, with the most significant association having a P-value of 0.004 with an odds ratio (OR) of 2.1. These two SNPs defined two associated SNP groups. Inheriting a minor SNP allele from both groups was a particular risk factor (OR=2.4, P=0.0008). Our data suggest that functional variants within the IL4R gene predispose to hip OA in Caucasian females. PMID- 14745653 TI - Mechanisms of alveolar epithelial repair in acute lung injury--a translational approach. AB - In patients with acute lung injury (ALI) or acute respiratory distress syndrome (ARDS), extensive damage to the alveolar epithelial and endothelial barrier is observed, resulting in the influx of protein-rich oedema fluid into the air spaces. Efficient alveolar epithelial repair is crucial to ALI/ARDS patients' recovery. Future therapeutic strategies may therefore include acceleration of the epithelial repair process in the injured lung. However, a better understanding of the cellular and molecular mechanisms that promote alveolar epithelial repair is needed if novel therapeutic strategies are to be developed. Pulmonary oedema fluid from patients with ALI/ARDS and from patients with hydrostatic oedema as control was obtained, and the effect on alveolar epithelial repair in vitro using our alveolar epithelial wound repair bioassay was studied. In contrast to the initial hypothesis, pulmonary oedema fluid from ALI/ARDS patients increased alveolar epithelial repair in vitro by an interleukin-1beta (IL-1beta)-dependent mechanism, demonstrating a novel, possibly beneficial role for IL-1beta in patients with ALI/ARDS. Further studies using primary alveolar epithelial cells from rats revealed that IL-1beta induced alveolar epithelial repair by an epidermal growth factor (EGF)/transforming growth factor-alpha (TGF-alpha) dependent pathway. Besides EGF and TGF-alpha, keratinocyte growth factor (KGF) and hepatocyte growth factor (HGF)--both present in pulmonary oedema fluid obtained from patients with ALI/ARDS--stimulate alveolar epithelial repair in vitro. Further experimental and clinical studies will show whether acceleration of alveolar epithelial repair by modulating cytokines and growth factors in the injured lung represents a promising new therapeutic strategy in patients with ALI/ARDS. PMID- 14745654 TI - Appropriateness of serum level determinations of antiepileptic drugs. AB - AIM: To assess the appropriateness of the determination of the serum levels of the antiepileptic drugs (AEDs) phenytoin, valproic acid and carbamazepine in inpatients of a tertiary care institution. METHODS: We performed a retrospective analysis of 602 AED serum level determinations. Appropriateness criteria regarding indication and timing were defined a priori using existing criteria from the literature. The main outcome measure was the proportion of serum levels with an appropriate indication and sampling time. RESULTS: Of 602 levels assessed, 463 (77%; 95% confidence interval [95% CI], 74-80%) had an appropriate indication with a range of 68% to 84% for individual AEDs; overall, 65% (95% CI: 61-69%) met the criteria for appropriate timing. Combining the two criteria, 268 (48%; 95% CI: 44-52%) AED level measurements were assessed as appropriate. Of 139 (23%, 95% CI: 20-27%) levels assessed as having an inappropriate indication, the majority (77%) were performed for routine monitoring. CONCLUSIONS: Less than half of all AED measurements met our criteria for appropriate AED level determinations. This creates unnecessary costs. Our data indicate the need for means to improve the rational use of AED serum level determination. PMID- 14745655 TI - Prevalence rate and reasons for refusals of influenza vaccine in elderly. AB - More knowledge on the reasons for refusal of the influenza vaccine in elderly patients is essential to target groups for additional information, and hence improve coverage rate. The objective of the present study was to describe precisely the true motives for refusal. All patients aged over 64 who attended the Medical Outpatient Clinic, University of Lausanne, or their private practitioner's office during the 1999 and 2000 vaccination periods were included. Each patient was informed on influenza and its complications, as well as on the need for vaccination, its efficacy and adverse events. The vaccination was then proposed. In case of refusal, the reasons were investigated with an open question. Out of 1398 patients, 148 (12%) refused the vaccination. The main reasons for refusal were the perception of being in good health (16%), of not being susceptible to influenza (15%), of not having had the influenza vaccine in the past (15%), of having had a bad experience either personally or a relative (15%), and the uselessness of the vaccine (10%). Seventeen percent gave miscellaneous reasons and 12% no reason at all for refusal. Little epidemiological knowledge and resistance to change appear to be the major obstacles for wide acceptance of the vaccine by the elderly. PMID- 14745656 TI - Perioperative management of patients with increased risk of laparoscopy-induced hepatic hypoperfusion. AB - Because hepatic hypoperfusion induced by laparoscopy has been underestimated, the aim of this article is to review the numerous factors influencing hepatosplanchnic blood flow during laparoscopy and to alert clinicians to the adverse consequences of hepatic hypoperfusion in high risk patients undergoing this procedure. PMID- 14745657 TI - Optimising the performance of an outpatient setting. AB - BACKGROUND: An outpatient setting typically includes experienced and novice resident physicians who are supervised by senior staff physicians. The performance of this kind of outpatient setting, for a given mix of experienced and novice resident physicians, is determined by the number of senior staff physicians available for supervision. The optimum mix of human resources may be determined using discrete-event simulation. METHODS: An outpatient setting represents a system where concurrency and resource sharing are important. These concepts can be modelled by means of timed Coloured Petri Nets (CPN), which is a discrete-event simulation formalism. We determined the optimum mix of resources (i.e. the number of senior staff physicians needed for a given number of experienced and novice resident physicians) to guarantee efficient overall system performance. RESULTS: In an outpatient setting with 10 resident physicians, two staff physicians are required to guarantee a minimum level of system performance (42-52 patients are seen per 5-hour period). However, with 3 senior staff physicians system performance can be improved substantially (49-56 patients per 5 hour period). An additional fourth staff physician does not substantially enhance system performance (50-57 patients per 5-hour period). CONCLUSION: Coloured Petri Nets provide a flexible environment in which to simulate an outpatient setting and assess the impact of any staffing changes on overall system performance, to promote informed resource allocation decisions. PMID- 14745659 TI - Frequency and outcome of in-hospital resuscitation outside the ICU-setting. AB - BACKGROUND: Guidelines on performing cardiopulmonary resuscitation and its research have been published. Only few data concerning in-hospital resuscitation are available from Switzerland. The aim of our study was to evaluate the frequency and outcome of cardiopulmonary arrests in our hospital and to look for ways of improving our resuscitation management. METHODS: The prospective study was performed in the Kantonsspital Liestal, a primary care hospital with 360 beds, where about 24'300 in-patients were treated during the 2 year observation period. Only in-hospital resuscitations outside the ICU were included and recorded according to the Utstein criteria. RESULTS: Within a 24 months period, 61 emergency calls were registered. 25 patients needed cardiopulmonary resuscitation. Initial cardiac rhythms were available for all subjects: 8 patients had asystole, 7 ventricular fibrillation and 10 pulseless electrical activity. 12 of 25 resuscitated patients had a return of spontaneous circulation, 7 lived longer than 24 hours and 6 patients (24%) survived to hospital discharge, 4 of them in a very good or good neurological condition. After 12 months 3 patients (12%) were living independently at home, 2 patients had to be treated in a nursing home and 1 patient had died. CONCLUSIONS: Our data correspond to survival rates in larger studies from abroad but are limited by the number of patients investigated. Improvements are necessary in documentation of resuscitation efforts. Rapid defibrillation must be further stressed. The implementation of a multicentre study is suggested because quality control and further improvement of in-hospital resuscitation are needed in Switzerland. PMID- 14745660 TI - Where we are--where we are heading. PMID- 14745658 TI - Salmeterol/fluticasone propionate (50/250 microg) combination is superior to double dose fluticasone (500 microg) for the treatment of symptomatic moderate asthma. AB - QUESTIONS UNDER STUDY: if patients with asthma remain symptomatic in spite of chronic treatment with inhaled corticosteroids (ICS), increasing the ICS dosage or adding another drug to the treatment regimen are possible therapeutic alternatives. We compared the efficacy and safety of combined salmeterol fluticasone therapy (SFC, 50/250 microg twice daily) with twice the dose of fluticasone propionate (FP, 500 microg b.i.d.) in symptomatic asthmatics. METHODS: this prospective, double-blind study was conducted in 76 study centres. 365 symptomatic patients with moderate asthma aged >18 years and receiving ICS in a dose equivalent to 1000 microg beclomethasone propionate per day were randomly assigned to receive either salmeterol xinafoate (50 microg) and fluticasone propionate (250 microg) in a single dry powder inhaler (Diskus) or 500 microg FP twice daily. The primary efficacy endpoint was morning peak expiratory flow rate (PEFR). Secondary measurements included forced expiratory volume in 1 second (FEV1), asthma symptom scores, and use of rescue medication. RESULTS: combined salmeterol fluticasone therapy resulted in significantly greater improvements in PEFR and symptom control than doubling the dose of FP. At week 12, morning PEFR had increased by 52 L/min from baseline in patients on SFC and by 36 L/min in subjects receiving FP. The adjusted difference between groups was 16.6 L/min (95% confidence interval, 1.1 to 32.0 L/min). In the SFC group, the percentage of symptom-free days increased from baseline by 49% of days as compared with 38% of days after FP (adjusted difference: 12.6% of days, 95% CI 4.0 to 20.7). Quality of life improved to a greater degree after SFC therapy, and patients regarded study drugs as superior to their previous asthma medication. Adverse event profiles were similar between groups. CONCLUSIONS: the combination of salmeterol 50 microg and fluticasone 250 microg in a single dry powder inhaler was superior to twice the dose of FP (500 microg). It seems justified to add salmeterol rather than increasing the ICS dose if symptomatic asthmatics require supplementary therapy. PMID- 14745661 TI - Corticosteroid-dependent hypertension: environmental influences. AB - Hypertension is a major disease of later life affecting 25% of the adult population in the industrialized world with most hypertensive individuals diagnosed as having essential hypertension. Approximately one third of these patients have elevated blood pressure due to increased sodium and water retention by the kidney resulting in suppressed plasma renin and aldosterone concentrations with high urinary potassium excretion and low plasma potassium levels. Monogenic forms of corticosteroid-dependent hypertension are rare. However, the discovery of these disorders has revealed genes whose proteins play a key role in the regulation of sodium homeostasis. The impaired function of these proteins, due to altered protein expression or the presence of inhibitors, contributes to the development of corticosteroid-dependent hypertension. This article focuses on the potential impact of environmental influences on corticosteroid-dependent regulation of sodium homeostasis and the development of hypertension. PMID- 14745662 TI - Intravenous thrombolysis for acute ischaemic stroke in a hospital without a specialised neuro-intensive care unit. AB - BACKGROUND: Treatment with intravenous (i.v.) recombinant tissue plasminogen activator (rt-PA) is recommended for selected patients with acute ischaemic stroke. We evaluated the feasibility and safety of this treatment in clinical practice in a hospital without a specialised neuro-intensive care unit. METHODS: We prospectively studied all patients who were treated with i.v. rt-PA for ischaemic stroke at our hospital between January 2001 and June 2002. The selection criteria corresponded to those published by the NINDS [1] and ECASS [2] groups. Time intervals between stroke symptom onset, hospital arrival and treatment with rt-PA were measured. A modified NIH stroke scale was used to assess clinical outcome 24 hours after stroke onset and before discharge. Cerebral computed tomography was performed prior to thrombolysis and again if the neurological status failed to improve or deteriorated. RESULTS: Thrombolytic therapy was administered to 15 acute ischaemic stroke patients, 13 men and two women with a median age of 69 years. The median time from stroke onset to rt-PA therapy was 135 minutes and from arrival in the emergency room to the start of thrombolysis 74 minutes. Ten patients exhibited early clinical improvement, defined as a decrease in NIHSS score by 4 points at 24 hours. Further improvement until discharge was observed in nine of these ten patients. One patient developed a non-fatal intracerebral haemorrhage. Another patient with severe stroke and clinical failure of thrombolysis died after 25 days. CONCLUSIONS: This study in a small patient population suggests that thrombolysis with rt-PA for acute ischaemic stroke is feasible without excess risk in a hospital experienced in the management of stroke patients, with a neurological consultant service but without a specialised neuro-intensive care unit (NICU). The outcome in this small series of patients corresponds to the results described in the randomised trials. PMID- 14745663 TI - Ten years of lung transplantation in Switzerland: results of the Swiss Lung Transplant Registry. AB - OBJECTIVE: Lung transplantation has evolved from an experimental procedure to a viable therapeutic option in many countries. In Switzerland, the first lung transplant was performed in November 1992, more than ten years after the first successful procedure world-wide. Thenceforward, a prospective national lung transplant registry was established, principally to enable quality control. PATIENTS: The data of all patients transplanted in the two Swiss Lung Transplant centres Zurich University Hospital and Centre de Romandie (Geneva-Lausanne) were analysed. RESULTS: In 10 years 242 lung transplants have been performed. Underlying lung diseases were cystic fibrosis including bronchiectasis (32%), emphysema (32%), parenchymal disorders (19%), pulmonary hypertension (11%) and lymphangioleiomyomatosis (3%). There were only 3% redo procedures. The 1, 5 and 9 year survival rates were 77% (95% CI 72-82), 64% (95% CI 57-71) and 56% (95% CI 45-67), respectively. The 5 year survival rate of patients transplanted since 1998 was 72% (95% CI 64-80). Multivariate Cox regression analysis revealed that survival was significantly better in this group compared to those transplanted before 1998 (HR 0.44, 0.26-0.75). Patients aged 60 years and older (HR 5.67, 95% CI 2.50-12.89) and those with pulmonary hypertension (HR 2.01, 95% CI 1.10-3.65) had a significantly worse prognosis The most frequent causes of death were infections (29%), bronchiolitis obliterans syndrome (25%) and multiple organ failure (14%). CONCLUSION: The 10-year Swiss experience of lung transplantation compares favourably with the international data. The best results are obtained in cystic fibrosis, pulmonary emphysema and parenchymal disorders. PMID- 14745664 TI - Reducing the 137Cs-load in the organism of "Chernobyl" children with apple pectin. AB - As a complement of standard radioprotective measures, apple-pectin preparations are given, especially in the Ukraine, to reduce the 137Cs uptake in the organism of children. The question has been raised: is oral pectin also useful when children receive radiologically clean food, or does this polysaccharide only act in binding 137Cs in the gut, blocking its intestinal absorption? In this case, pectin would be useless if radiologically clean food could be given. The study was a randomised, double blind placebo-controlled trial comparing the efficacy of a dry and milled apple-extract containing 15-16% pectin with a similar placebo powder, in 64 children originating from the same group of contaminated villages of the Gomel oblast. The average 137Cs load was of about 30 Bq/kg bodyweight (BW). The trial was conducted during the simultaneous one-month stay in the sanatorium Silver Spring. In this clean radiological environment only radiologically "clean" food is given to the children. The average reduction of the 137Cs levels in children receiving oral pectin powder was 62.6%, the reduction with "clean" food and placebo was 13.9%, the difference being statistically significant (p <0.01). The reduction of the 137Cs load is medically relevant, as no child in the placebo group reached values below 20 Bq/kg BW (which is considered by Bandazhevsky as potentially associated with specific pathological tissue damages), with an average value of 25.8 +/- 0.8 Bq/kg. The highest value in the apple-pectin group was 15.4 Bq/kg, the average value being 11.3 +/- 0.6 Bq/kg BW. PMID- 14745665 TI - A suprasellar arachnoid cyst destructing sphenoid sinus: an unusual cause of headache in an elderly female. PMID- 14745666 TI - Cheyne-Stokes respiration in patients with congestive heart failure. AB - Cheyne-Stokes Respiration (CSR) is a breathing pattern characterised by rhythmic oscillation of tidal volume with regularly recurring periods of hyperpnoea, hypopnoea and apnoea. CSR is no longer solely regarded as a symptom of severe congestive heart failure (CHF), but has been recognised as an independent risk factor for worsening heart failure and reduced survival in patients with CHF. CSR is associated with frequent awakening that fragment sleep and with concomitant sympathetic activation both of which may worsen CHF. Cheyne-Stokes Respiration is very common in patients with severe CHF and its prevalence may have been underestimated in the past due to technical limitations that precluded respiratory monitoring outside sleep laboratories. Since treatment of CSR appears to be beneficial and safe, patients at risk should be promptly diagnosed and treated. Treatment of CSR has been demonstrated to improve left ventricular ejection fraction and potentially prolongs survival in patients with severe CHF. This article briefly summarises the current knowledge of the patho-physiology, prevalence and therapy of Cheyne-Stokes respiration. PMID- 14745667 TI - The susceptibility of patients with type-2 diabetes to hepatitis C virus infection during long-term haemodialysis. AB - BACKGROUND: Type-2 diabetes has emerged as the commonest cause of end stage renal disease (ESRD) requiring long-term haemodialysis (HD) that constitutes a high risk environment for the transmission of hepatitis C virus (HCV). The likelihood of acquiring HCV infection in this rapidly growing population on HD conceivably vulnerable to viral infections has not been well studied. The present study aims to determine the susceptibility of the patients with type-2 diabetes to HCV infection in a HD unit with high HCV prevalence. METHODS: The records of 196 patients with ESRD enrolled on long-term HD at King Fahad Hospital and tertiary care centre, in Hofuf, Saudi Arabia, from November1995 to November 2000, were retrospectively reviewed. HCV prevalence, seroconversion rates, history of blood transfusion, and time on dialysis (time span since initiation of HD therapy) were recorded and compared between the group of patients with type-2 diabetes, and the non-diabetic group. RESULTS: The overall, HCV seroprevalence of 41.3% (81/196) and annual seroconversion rate of 8.26% were observed. Anti-HCV positivity was associated with longer time on dialysis. Of the 196 patients 54 (27.5%) had type 2 diabetes mellitus and 142 (72.5%) were non-diabetics. Patients with type-2 diabetes recorded higher HCV prevalence (57.4% vs 35.2%), and annual seroconversion rates (11.48% vs 7.04%) after a shorter period on dialysis (32.6 vs 50.6 months), as compared to those of the non-diabetic group. CONCLUSIONS: A significantly higher HCV prevalence [odds ratio (OR)-2.462, 95% CI (1.338- 4.542)] and annual seroconversion rate [OR-2.483, 95% CI (1.241-4.946)] despite relatively shorter period on dialysis [OR-3.320, 95% CI (1.487-7.4810)] among patients with type-2 diabetes clearly point to the greater likelihood of their acquiring HCV infection even at an earlier stage than the non-diabetic patients, receiving treatment in a high prevalence HD unit. PMID- 14745668 TI - 20 years of peritoneal dialysis in a mid-sized Swiss hospital. AB - PRINCIPLES: Few long term studies exist about peritoneal dialysis (PD). We collected the experiences over nearly 20 years in a single mid-sized centre in Switzerland. METHODS: In a retrospective survey we examined our PD-cohort with respect to mortality, technique survival, peritonitis rate and other complications. We calculated the proportion of PD-patients of the total dialysis population (penetration rate) and measured the time of PD-associated hospitalisations. RESULTS: 50 patients were included during an observation period of 20 years. The mean penetration rate was 23% (range 11% to 34%). The mean treatment time per patient was 2.8 years (median: 3.6 years; range 0.4-9.5 years). Patient survival was 80% at three years and 60% at five years. Technique retention rate was 40% after three, and 20% after five years. Each of the three outcome categories--transplantation, switch to haemodialysis (HD) and death during PD--accounted for one third of the PD drop-out number. CONCLUSION: Compared to the average of Swiss dialysis centres the penetration rate is high. Patient and technique survival correspond to data in the literature, as do the frequency and types of complications. We consider PD as an efficient and well tolerated dialysis modality, which should be offered also in smaller dialysis centres. Since PD is not only feasible, but appears to be less costly than HD, we recommend PD as the first-line dialysis option for patients in end-stage renal disease. PMID- 14745669 TI - Facial features and skeletal abnormalities in Larsen syndrome--a study of three generations of a Tunisian family. AB - We report on a 3 generation study of a Tunisian family, in which eight subjects had or have features of Larsen syndrome: three siblings, two females and one male are affected with flattened facies, multiple congenital joint dislocations, and club foot deformities. Five other family members were recognised as being variably affected with the syndrome. Over the three generations, despite the characteristic facial features being the most constant clinical signs of the syndrome, none of those still living had palatal clefts, the multiple infantile deaths in this family, however, were characterised by an association with cleft palate. PMID- 14745670 TI - Stressors and strains of medical training and practice. PMID- 14745671 TI - Notice of retraction. PMID- 14745672 TI - [Lamellar kreatoplasty - new concepts]. PMID- 14745673 TI - [Lamellar keratoplasty - new concepts]. AB - New concepts of lamellar keratoplasty techniques mainly deal with the refinement of the so-called deep lamellar endothelial keratoplasty (DLEK). The aim of the deep lamellar technique is to expose bare Descemet's membrane of the recipient and to suture in a full thickness graft. Graft rejections are not known with DLEK. We describe the preparation techniques of Anwar, Melles, and Krumeich and discuss the advantages and disadvantages. The femtosecond laser enables the surgeon to cut the cornea non-mechanically with a cutting accuracy of +/- 10 microns. Fuchs endothelial dystrophy can be treated by posterior lamellar keratoplasty (POLK). In this technique only a sheet of Descemet's and endothelium are replaced. In severe alkali burn with limbal stem cell deficiency a lamellar corneo-scleral disc can be grafted in order to restore the limbus. Routine central penetrating keratoplasty should follow at least half a year later. PMID- 14745674 TI - [Subconjunctival anaesthesia using cocaine or mepivacaine for cyclocryotherapy in advanced glaucoma]. AB - BACKGROUND: Retro- or peribulbar anaesthesia are the standard procedures for cyclodestructive surgery. Because these methods of anaesthesia may further compromise optic nerve function, especially in advanced glaucoma, subconjunctival anaesthesia was evaluated as an alternative procedure in cyclocryotherapy. PATIENTS AND METHODS: 1. To evaluate the current methods of anaesthesia a questionnaire was sent to all University Eye Clinics in Germany in 2001. 2. A prospective, consecutive study concerning cyclocryotherapy in advanced glaucoma was started using subconjunctival anaesthesia with 5 % cocaine or, alternatively, 2 % mepivacaine in 25 eyes of 25 patients in each group. Pain during cyclocryotherapy was recorded using a 10-point rating-scale (1 = no pain to 10 = intolerably severe pain) and adverse events were also recorded. RESULTS: 1. 88.6 % of the University Eye Clinics in Germany replied to our questionnaire. In 2001 cyclocryotherapy was performed in 80.6 % of the University Eye Clinics in Germany (Tubingen not included), 16.1 % among them solely used cyclocryotherapy for the treatment of advanced glaucoma. 2. Using 5 % cocaine for subconjunctival anaesthesia - 92 % of patients experienced no pain and 8 % of patients reported very mild pain during cyclocryotherapy. One patient reported nausea, 4 patients mouth dryness. After anaesthesia with mepivacaine - 80 % experienced no pain, 12 % very mild pain, and 8 % mild pain. There were no systemic side effects. CONCLUSION: Most of the patients experienced no pain during cyclocryotherapy using subconjunctival anaesthesia with cocaine or mepivacaine. Therefore, risks and side effects of retro- or parabulbar anaesthesia can be successfully avoided by this simple modification. PMID- 14745675 TI - [Frequency of reoperations after amniotic membrane transplantation]. AB - BACKGROUND: Retrospective evaluation of the frequency of reoperations after amniotic membrane transplantation for different pathologic entities. MATERIAL AND METHODS: We included 81 cases of amniotic membrane transplantation (AMT), which were operated on at the university eye clinic Tubingen since 1997 and which had been followed up for at least 12 months. Patient diagnoses were subdivided as follows: conjunctival defects after tumour excision (N = 2), bullous keratopathy (N = 5), external fistula after glaucoma filtering surgery (N = 3), recurrent pterygium (N = 5), symblepharon (N = 6), corneal ulcer with descemetocele (N = 3), non-perforated corneal ulcer (N = 51), perforated corneal ulcer (N = 1), other (n = 5). RESULTS: The overall reoperation rate was determined to be 42 % after a follow-up period of one year. Most frequently, a second AMT (N = 16) and a perforating keratoplasty (N = 16) were performed after the initial AMT. The following reoperation rates were determined for the subgroups: Conjunctival defects after tumour excision (0 %), Bullous keratopathy (60 %), External fistula after glaucoma filtering surgery (67 %), Recurrent pterygium (60 %), Symblepharon (67 %), Corneal ulcer with descemetocele (67 %), Non-perforated corneal ulcer (49 %), Perforated corneal ulcer (100 %). CONCLUSIONS: Particularly in patients with conjunctival defects after tumour excision and with non-perforated corneal ulcers, stabilisation of the ocular surface homeostasis can be achieved with a single amniotic membrane transplantation for at least one year. PMID- 14745676 TI - [Laser in situ keratomileusis with a flying-spot excimer laser and a carriazo barraquer microkeratome - outcomes after 6 months]. AB - PURPOSE: To evaluate safety, efficacy, predictability and refractive stability of LASIK, using a flying-spot excimer laser, 91 eyes were examined during a 6-months follow-up. PATIENTS AND METHODS: Consecutively, 91 eyes were enrolled in the study, each eye previously underwent LASIK with the ESIRIS excimer laser (ESIRIS, supplier: eye-tech-solutions Schwind, Kleinostheim, Germany). 91 myopic eyes (mean preoperative refraction: - 5.4 D, range: - 3 to - 13.3 D, astigmatism: - 1 D, range: 1.3 to 3.5 D) were included. A full ophthalmological examination was performed during the first week, 1, 3 and 6 months postoperatively. RESULTS: 26 % of all eyes achieved an uncorrected visual acuity of 0.8, 35 % of 1.0 and 5 % 1.2 6 months postoperatively. 23 % of the eyes lost 1 line of best spectacle corrected visual acuity (BSCVA), 25 % gained one or more lines of BSCVA. Refraction of 90 eyes were within +/- 1 D of emmetropia 6 months postoperatively. There was no change in spherical equivalent within the interval of 6 months, all eyes were within +/- 0.5 D. CONCLUSION: LASIK with the ESIRIS excimer laser was an effective and safe option with good stability and predictability within the specified interval. PMID- 14745677 TI - [Heparin Surface-Modified Poly(methylmethacrylate) and Foldable Hydrophobic Acrylic Intraocular Lenses in Cataract Patients with Acquired Immune Deficiency Syndrome and CMV-Retinitis]. AB - BACKGROUND: In HIV-positive patients with necrotising retinitis, well-tolerated lens material as well as minimal invasive surgery to avoid postoperative complications are imperative. Heparin surface-modified PMMA-IOLs as well as acrylic IOLs are associated with less postoperative inflammation than conventional intraocular lenses. METHODS: In this randomised prospective clinical trial 18 patients received 7 HSM-IOLs through a self-sealing scleral tunnel and 11 foldable Acrysof-IOLs through a self-sealing limbal tunnel incision by the same surgeon. 7 of the 18 eyes were silicone oil-filled before cataract surgery, one eye received oil after the IOL implantation. Visual acuity, intraocular pressure, contrast sensitivity, intraocular inflammation and posterior capsule opacification were assessed. The follow-up period was 1 year. RESULTS: Eyes with Acrysof-IOLs were associated with lower, but not statistically significant, laser flare photometry values (photon counts/ms) than those with HSM-IOLs pre- (90.18 +/- 54.7 vs. 73.92 +/- 6.44) and 3 months post-surgery (69.06 +/- 8.27 vs. 55.03 +/- 7.75). 6 of the 7 eyes with HSM-IOL and 3 eyes of the Acrysof-group developed a dense posterior capsule opacification (PCO), 2 eyes had no and 6 eyes only a distinct PCO. YAG capsulotomy did not improve visual acuity in any case but retinal examination was again possible. Eyes with Acryl-IOL showed pre- and postoperatively a better visual acuity than eyes with HSM-IOL ("hand movement" - 0.4 vs. "no light perception" - 0.1 pre- and 0.05 - 0.6 vs. "no light perception" - 0.1 3 months postoperatively. Contrast sensitivity testing (Vistech method) could be performed in the PMMA-group preoperatively only in 1 eye (A1), in the Acryl-group in 2 eyes (A4, B4, C2, D2) and revealed postoperatively "no contrast vision" - A1, B2, C1 vs. "no contrast vision" - A2, B2, C3, D1, E1. The intraocular pressure was preoperatively 13.86 mm Hg (PMMA) vs. 14.82 mm Hg (Acrysof) and 14.4 mm Hg vs. 12.89 mm Hg 3 months post surgery. CONCLUSION: In the parameters we observed there was no statistical significant difference between heparin-surface modified (Pharmacia 811 C) and hydrophobic acrylic IOLs (Alcon Acrysof MA60BM) in patients with AIDS. As foldable Acrysof-IOLs can be implanted through a self-sealing bloodless tunnel incision and are associated with less posterior capsule opacification we prefer this IOL type in cataract surgery of AIDS patients. PMID- 14745678 TI - [Diffuse diabetic macular edema - a therapeutic challenge]. PMID- 14745679 TI - [Grid laser photocoagulation in diffuse diabetic macular edema]. AB - BACKGROUND: To evaluate the clinical outcome of macular grid laser photocoagulation in the treatment of diffuse diabetic macular oedema. PATIENTS AND METHOD: The retrospective study included 30 consecutive patients (41 eyes) who were treated by macular argon green grid laser photocoagulation for diffuse diabetic macular oedema. Follow-up time was 31.4 +/- 19.6 weeks. RESULTS: Visual acuity decreased from 0.25 +/- 0.18 (range, 0.03 - 0.8) to 0.20 +/- 0.18 (range, 0.02 - 0.8) (P = 0.045), representing a change of - 0.9 +/- 2,32 lines. 5 (12.2 %) eyes gained in visual acuity, visual acuity remained unchanged for 23 (56.1 %) eyes, and 13 (31.7 %) eyes showed a visual loss of more than one line. In eyes with a baseline visual acuity > or = 0.2 (N = 24) visual acuity dropped from 0.36 +/- 0.15 (0.2 - 0.8; median 0.3) to 0.29 +/- 0.19 (0.05 - 0.8; median 0.2) (p = 0.038). 3 eyes (12.5 %) gained > or = 2 lines, 11 eyes (45.8 %) lost > or = 2 lines, 10 eyes (41.7 %) remained stable. Mean loss was - 1.63 +/- 2.53 lines. Eyes with a baseline visual acuity < or = 0.2 did not change significantly. CONCLUSION: In the present study mean visual acuity decreased in the whole population and especially in the subgroup with a baseline visual acuity of > or = 0.2 after macular grid laser photocoagulation for diffuse diabetic macular oedema. Mean visual loss was just below the predefined 2 lines. In view of these results and upcoming new pharmacological and surgical treatment modalities, the significance of grid laser photocoagulation should be re-discussed. PMID- 14745680 TI - [Nosocomial pseudomonas aeruginosa-associated keratitis in soft contact lens wearer]. AB - BACKGROUND: Pseudomonas aeruginosa is the most common cause of bacterial associated keratitis in soft contact lens wearers, due to wrong use of soft contact lenses. Problems are often severe corneal ulcers and even corneal perforations. We report on a soft contact lens wearer with credibly correct use of soft contact lenses and nosocomial Pseudomonas aeruginosa-associated keratitis. CASE REPORT: A 33-year old woman suffered from corneal ulcer and corneal infiltration with beginning endophthalmitis 2 days after having used of new soft contact lenses. After systemic and local antibiosis and penetrating keratoplasty we could stop endophthalmitis before reaching the vitreous and retina. RESULTS: Histological and microbiological examinations showed a corneal ulcer with severe corneal infection due to Pseudomonas aeruginosa with resistance to mezlocillin and intermediale resistance to gentamicin. After therapy a stable situation with visual acuity of 20/60 was attained. CONCLUSIONS: Previous reports on Pseudomonas aeruginosa-associated keratitis in soft contact lens wearers demonstrate corneal problems due to extended or overnight wear or unsuccessful contact lens cleaning. We present a case of nosocomial corneal infection after soft contact lens wearing and nosocomial infection because of contact with a partner working in an intensive-care unit. Hygienic rules should be strictly followed by patients and staff using soft or hard contact lenses for visual correction or for therapeutic reasons. PMID- 14745681 TI - [The "Clothesline Phenomenon" after Cerclage with Arruga's Suture - Is Cutting of the Suture Necessary?]. AB - BACKGROUND: Several early and late complications have been described since the introduction of cerclage surgery using a supramid suture. Due to cutting of the supramid suture through scleral tissue the supramid suture may appear at the fundus. Long-term results of scleral penetration and crossing the bulbus by the supramid suture, which is then only covered by the retina ("clothesline phenomenon"), could not be described so far. We report on a patient 20 years after cerclage surgery with Arruga's suture. CASE REPORT: A now 63-year old female patient with high myopia (D - 14.0 OU) suffering from complete retinal detachment had had ophthalmic surgery using Arruga's encircling suture with additional horizontal buckling at her left eye in 1983. Postoperative visual acuity was 10/20. The retina was attached. In 2002 phakoemulsification with posterior chamber lens implantation due to cataracta corticalis was done externally. In 2003 a complete scleral and chorioidal penetration and appearance of the Arruga's suture behind the retina occurred extending from 4 over 6 to 8 o'clock position with a visual acuity of 20/25. The patient did not agree with the cutting of the suture at any time. CONCLUSIONS: The "clothesline phenomenon" is a very rare late complication after encircling surgery using an Arruga suture by retinal detachment. Suture cutting is suggested because of a possible risk of retinal detachment. As shown in our case report, stable retinal attachment after Arruga's suturing is possible. PMID- 14745682 TI - [Heterochromia of the Iris after t-PA and Gas Injection of Sulfur Fluoride]. AB - BACKGROUND: Are changes in iris colour and retinal pigment epithelium after t-PA and gas injection and subsequent vitreous haemorrhage caused by blood cells, or is this a toxic effect? CASE: A 81-year old female presented for vitrectomy with a persistent vitreous haemorrhage two months after t-PA and gas injection because of subretinal macular haemorrhage. Sonographic control revealed attached retina. The examination disclosed a new heterochromia of the iris with a change from blue to green-brown. A discrete anterior chamber flare and blood cells were found. Subsequent vitrectomy revealed a thick, rubber-like haemorrhage in the vitreous with suspect pigmentation. A change in retinal pigmentation was also evident. CONCLUSION: The cause for the heterochromia and the change in pigmentation of the retinal pigment epithelium remained unclear. It may have been a consequence of the persistent bleeding with iron apposition on the iris. A toxic effect of t-PA has to be discussed which led to the alteration in pigmentation. PMID- 14745683 TI - [Reversible Perimacular Retinopathy by Long-Time Interferon alpha Therapy]. AB - BACKGROUND: Recently interferon therapy has become increasingly important in the treatment of viral and malignant diseases. Subsequently, a variety of adverse effects of interferon use has been reported. Ocular complications following interferon therapy include retinal ischaemia with cotton wool spots, retinal haemorrhages, capillary nonperfusion, vasospasm, and arteriolar occlusion. CASE REPORT: A 37-year old male complained about increasingly blurred vision in his right eye. Because of a chronic type B hepatitis he has been treated with interferon alpha for a total of 35 weeks. Visual acuity was slightly reduced (OD cc 0.9, OS cc 1.0 p). Dilated fundus examination and fluorescein angiography showed an incipient ischaemic retinopathy with perimacular arteriolar alterations in both eyes and single cotton wool spots in his right eye. Perimetry showed a slight reduction of the mean deviation. Amsler-grid did not show any pathology. As the retinopathy was thought to be related to the interferon treatment, it was decided to discontinue this therapy immediately. 2 weeks later visual acuity was improved (OD cc 1.25 p, OS cc 1.0) and dilated fundus examination showed no signs of perimacular ischaemic retinopathy and the initial cotton wool spots had been resolved. CONCLUSION: In patients with long-time interferon treatment non specific visual complaints like blurred vision or dark spots might be symptoms of an early ischaemic retinopathy. Retinal alterations might be reversible if treatment is discontinued. If an ischaemic retinopathy is present it should be considered to abandon interferon therapy, since permanent loss of vision has been described in advanced capillary non-perfusion and arteriolar occlusion. However, the decision to discontinue the interferon therapy should be made interdisciplinary while considering diagnosis and therapy indication. PMID- 14745684 TI - [Stigma experiences of patients with obsessive compulsive disorders]. AB - BACKGROUND: Only in recent years has research shown interest in stigma experiences of patients with mental disorders. However, subjective experiences of patients with OCD have not been published until now. METHOD: Fifty-one patients with OCD were questioned about anticipated and actual stigmatization experiences as well as their stigma coping strategies, using a questionnaire especially developed for this study. RESULTS: Most of the patients expected negative reactions from their environment. Eighty per cent of the respondents were convinced that they would be rejected at work when it became known that they are mentally ill. Two thirds of the respondents feared that they would be rejected in a partnership. Concrete stigmatization was rarely reported by patients with OCD. Contrary to patients with depression and schizophrenia, the group of patients with OCD showed some differences between anticipated and actually experienced stigmatization. Concealing is a relevant strategy for dealing with the illness for three quarters of the respondents. CONCLUSION: Possibilities of reducing stigmatization and discrimination because of mental illness in general and OCD in particular are discussed. PMID- 14745685 TI - [Is premature termination of opiate detoxification due to intensive withdrawal or craving?]. AB - Opiate addicts terminate inpatient detoxification prematurely in about 50% of treatment episodes. Premature termination of treatment is often considered to be motivated by intensive withdrawal symptoms. Therefore, the relation between discontinuing treatment and the intensity of withdrawal symptoms and heroin craving is investigated. 130 opiate addicts consecutively admitted to a detoxification ward daily assessed the intensity of withdrawal symptoms on the Short Opiate Withdrawal Scale (SOWS) as well as the intensity of heroin craving on a visual analogue scale. Withdrawal symptoms were treated by stepwise reduction of methadone and symptom-oriented medication. 66 patients (50.8%) terminated treatment prematurely. However, during the days preceding treatment termination, these patients did not differ from regularly detoxified patients assessed on corresponding days with respect to craving, and reported even less intensity of withdrawal symptoms. In conclusion, neither the intensity of withdrawal symptoms nor the intensity of heroin craving constitutes the primary reason for premature termination of detoxification. PMID- 14745686 TI - [Kleptomania: clinical and legal aspects]. AB - Kleptomania is characterized by repetitive and irresistible impulses to steal objects, with growing excitement before, and relief after the incidents. The thefts are not planned, carried out without confidants and regarded as illegal by the offenders who often feel guilty for it. The objects are stolen not for monetary gain or personal use, but are mostly discarded, given away or hoarded. Though there is a debate going on for more than 100 years, the nosological position of kleptomania remains unclear. On the one hand the clinical picture is observed as an isolated disorder in otherwise healthy persons, on the other hand it may also occur as an accompanying symptom of various other psychiatric disorders, e. g. impulse control disorders, acute psychoses, affective, or organic psychic disorders. The present article gives an overview of the most important aetiopathogenetic viewpoints about kleptomania and discusses their nosological, therapeutical and legal implications. PMID- 14745687 TI - [Spongiform leucoencephalopathy after inhaling illicit heroin and due to carbon monoxide-intoxication]. AB - A spongiform leucoencephalopathy sometimes develops as a result of inhaling illicit heroin as well as due to carbon monoxide-intoxication. Clinically psychiatric symptoms precede a neurological deterioration. Some patients die. After a brief description of several epidemiological and historical-cultural aspects regarding the smoking of opiates, the typical neuroradiological signs such as hypodensity of the white matter in CCT and signal alterations in MRT, and neuropathological sequelae such as intramyelinic vacuolisation are listed. Pathophysiologically an edema of the white matter in the beginning is suspected. Second, a dysfunction of the mitochondria is addressed relying on the particular metabolism of the oligodendrocytes. Since smoking of heroin is an increasingly preferred way of application in all continents and therapeutic options are still lacking, the need of further explanation of the underlying processes is stressed. PMID- 14745688 TI - [Why are stereotypes about mentally ill so resistant? Lessons from social psychology]. AB - Prejudices and social stigma in mental illness have a long tradition and have hardly been modified by educational programmes on the long term. The question, why these negative social attitudes appear to be considerably resistant to change is analysed with reference to social psychological research on stereotypes. First, a short review of studies on public attitudes towards the mentally ill and media analyses will give evidence to the widespread stereotypical misconceptions and their reproduction by selected media information. Then the dynamics of social stereotyping will be explained in relation to cultural, cognitive and emotional factors focussing on the development and activation of stereotypes within the paradigm of information processing. It is supposed that stereotype activation facilitates the processing of consistent information as well as it inhibits the processing of inconsistent information. Implications for changing negative attitudes about the mentally ill are discussed. PMID- 14745689 TI - [Myasthenia gravis: pathology, diagnosis and treatment]. PMID- 14745691 TI - Prospects for preventing serious systemic toxemic complications of Shiga toxin producing Escherichia coli infections using Shiga toxin receptor analogues. PMID- 14745692 TI - Oral therapeutic agents with highly clustered globotriose for treatment of Shiga toxigenic Escherichia coli infections. AB - Shiga toxin (Stx) is a major virulence factor in infection with Stx-producing Escherichia coli (STEC). We developed a series of linear polymers of acrylamide, each with a different density of trisaccharide of globotriaosylceramide (Gb3), which is a receptor for Stx, and identified Gb3 polymers with highly clustered trisaccharides as Stx adsorbents functioning in the gut. The Gb3 polymers specifically bound to both Stx1 and Stx2 with high affinity and markedly inhibited the cytotoxic activities of these toxins. Oral administration of the Gb3 polymers protected mice after administration of a fatal dose of E. coli O157:H7, even when the polymers were administered after the infection had been established. In these mice, the serum level of Stx was markedly reduced and fatal brain damage was substantially suppressed, which suggests that the Gb3 polymers entrap Stx in the gut and prevent its entrance into the circulation. These results indicate that the Gb3 polymers can be used as oral therapeutic agents that function in the gut against STEC infections. PMID- 14745693 TI - Cerebrospinal fluid abnormalities in patients with syphilis: association with clinical and laboratory features. AB - OBJECTIVE: To define clinical and laboratory features that identify patients with neurosyphilis. METHODS: Subjects (n=326) with syphilis but no previous neurosyphilis who met 1993 Centers for Disease Control and Prevention criteria for lumbar puncture underwent standardized history, neurological examination, venipuncture, and lumbar puncture. Neurosyphilis was defined as a cerebrospinal fluid (CSF) white blood cell count >20 cells/ microL or reactive CSF Venereal Disease Research Laboratory (VDRL) test result. RESULTS: Sixty-five subjects (20.1%) had neurosyphilis. Early syphilis increased the odds of neurosyphilis in univariate but not multivariate analyses. In multivariate analyses, serum rapid plasma reagin (RPR) titer > or =1 : 32 increased the odds of neurosyphilis 10.85 fold in human immunodeficiency virus (HIV)-uninfected subjects and 5.98-fold in HIV-infected subjects. A peripheral blood CD4+ T cell count < or =350 cells/ microL conferred 3.10-fold increased odds of neurosyphilis in HIV-infected subjects. Similar results were obtained when neurosyphilis was more stringently defined as a reactive CSF VDRL test result. CONCLUSION: Serum RPR titer helps predict the likelihood of neurosyphilis. HIV-induced immune impairment may increase the risk of neurosyphilis. PMID- 14745694 TI - Increased burden of illness associated with antimicrobial-resistant Salmonella enterica serotype typhimurium infections. AB - This study investigated the burden of illness associated with 440 cases of Salmonella enterica serotype Typhimurium infection identified in Canada between December 1999 and November 2000. We categorized case subjects' infections by definitive phage type 104 (DT104) and antimicrobial-resistance patterns. These variables were then investigated as risk factors for hospitalization. Hospitalization was more likely to occur among case subjects whose infections were resistant to at least ampicillin, chloramphenicol and/or kanamycin, streptomycin, sulphamethoxazole, and tetracycline (R-type AK/CSSuT; odds ratio [OR], 2.3; P=.003), compared with case subjects with AK/CSSuT-susceptible infections, and among case subjects with non-DT104 R-type AKSSuT infections (OR, 3.6; P=.005), compared with case subjects with non-DT104 AKSSuT-susceptible infections. In contrast, hospitalization rates did not differ between case subjects with DT104 infections and case subjects with non-DT104 infections or between case subjects with DT104 R-type ACSSuT infections and case subjects with DT104 ACSSuT-susceptible infections. We estimated that 57% of the hospitalizations among AK/CSSuT case subjects and 72% of the hospitalizations among non-DT104 AKSSuT case subjects were attributable to the resistance patterns of the infections. PMID- 14745695 TI - Four antibiotic-resistant Streptococcus pneumoniae clones unrelated to the pneumococcal conjugate vaccine serotypes, including 2 new serotypes, causing acute otitis media in southern Israel. AB - This study examined the prevalence of antibiotic-resistant clones that belong to serotypes not included in the pneumococcal conjugate vaccines and that cause a significant percentage of acute otitis media (AOM) in children in southern Israel. During 1998-2001, 2467 pneumococcal isolates, obtained from middle-ear fluid of children <3 years old with AOM, were characterized by antimicrobial susceptibility testing, serotype testing, and pulsed-field gel electrophoresis. Non-vaccine type (NVT) strains constituted 477 (19%) of the 2467 isolates, of which 173 (36%) belonged to only 4 serotypes: 35B, 33F, 21, and 15B/C. For serotype 35B, 47 (96%) of 49 strains were penicillin nonsusceptible, and 93% constituted a single clone; for serotype 33F, 31 (82%) of 38 strains were penicillin nonsusceptible, and 95% constituted a single clone; for serotype 21, 38 (93%) of 41 strains were penicillin nonsusceptible, and 93% constituted a single clone; for serotype 15B/C, 22 (49%) of 45 strains were penicillin nonsusceptible, and 42% constituted a single clone. Two of these clones have not been described elsewhere. The high prevalence of NVT clones should increase the awareness of the potential for replacement of the vaccine strains with these NVT antibiotic-resistant strains. PMID- 14745696 TI - Inefficient replication of Listeria innocua in the cytosol of mammalian cells. AB - The efficiency of adherence to, internalization by, and replication in the cytosol of J774 macrophages and HEp-2 epithelial cells was compared between a nonspreading Listeria monocytogenes actA mutant and L. innocua. The studied L. innocua strains were equipped either with listeriolysin alone or with listeriolysin O (LLO) and the recently identified hexose-phosphate transporter of L. monocytogenes. All listerial strains expressed green fluorescent protein (GFP) under the control of the PrfA-dependent actA promoter. GFP expression was observed exclusively in the cytosol of host cells. Escape from the phagosome of LLO-expressing L. innocua strains was as efficient as that from L. monocytogenes. hpt-positive L. innocua showed significantly enhanced adherence to HEp-2 cells, but internalization was only slightly increased, compared with hpt-negative L. innocua. Subsequent replication of L. monocytogenes in the cytosol of the host cells proceeded within the next 6 h in most infected host cells, with a generation time <40 min. L. innocua prfA hly replicated more slowly (with a generation time of 60-90 min), and, in most host cells, bacterial replication stopped after 2-3 rounds of replication. In some cells, bacterial replication did not occur. Twenty-four hours after infection, the majority of J774 cells (>90%) infected with L. monocytogenes actA were dead, whereas most host cells infected with L. innocua were still alive. L. innocua equipped with the prfA, hly, and hpt genes of L. monocytogenes did not show significantly increased cytosolic replication, which indicates that expression of this sugar phosphate uptake system is not sufficient for extensive listerial replication in the cytosol of host cells. PMID- 14745697 TI - Neisseria meningitidis undergoes PilC phase variation and PilE sequence variation during invasive disease. AB - Neisseria meningitidis colonizes the upper respiratory tract (URT), enters the blood stream, and reaches the cerebrospinal fluid (CSF). In the present study, we show that bacteria isolated from the URT adhere better to human epithelial cells, compared with bacteria from blood or CSF, which suggests that important changes of virulence-associated proteins take place during bacterial dissemination. Phase variation in the pilus adhesin PilC and sequence variation in the pilus subunit PilE occurred among strains from 1 patient. Changes were not found in the invasion-associated opacity proteins or in lipooligosaccharides. PilC was frequently expressed in serogroup B strains and in URT strains but was often switched off in other serogroups and in CSF strains. Strains lacking PilC showed impaired adhesion to epithelial cells. These data argue that N. meningitidis undergoes PilC phase variation and PilE sequence variation during invasive disease. PMID- 14745698 TI - Inhibition of Neisseria gonorrhoeae genital tract infection by leading-candidate topical microbicides in a mouse model. AB - The development of effective vaginal microbicides is paramount in the fight against the spread of sexually transmitted infections. Preclinical testing of candidate microbicides for the prevention of gonorrhea has been seriously hindered by the lack of an animal model. We assessed the efficacy of 7 promising formulated agents--CarraGuard, Ushercell, [poly]sodium 4-styrene sulfonate (T PSS), PRO 2000, ACIDFORM, cellulose acetate phthalate (CAP), and BufferGel--by use of a mouse model of Neisseria gonorrhoeae genital tract infection. Mice received test agent, relevant placebo, or no treatment, followed by intravaginal N. gonorrhoeae challenge. N. gonorrhoeae colonization was tested by vaginal culture. CarraGuard, Ushercell, and T-PSS demonstrated significant protection, compared with control agents and no treatment. PRO 2000, ACIDFORM, and CAP showed significant protection, compared with no treatment but not compared with respective control agents. Mice that received BufferGel were provided significant protection, compared with untreated control mice; no placebo was tested. The findings of the present study suggest that topical agents may effectively reduce N. gonorrhoeae infection and that further evaluation is warranted. PMID- 14745699 TI - A putative sugar-binding transcriptional regulator in a novel gene locus in Enterococcus faecalis contributes to production of biofilm and prolonged bacteremia in mice. AB - A biofilm-negative transposon mutant was created from an Enterococcus faecalis strain that produces a lot of biofilm. The transposon had been inserted in the second gene of a locus consisting of 4 open-reading frames, designated bop (biofilm on plastic surfaces). A nonpolar deletion of this gene and of parts of the 2 flanking genes was created; production of biofilm by this deletion mutant was significantly enhanced, compared with that by the wild-type strain. Expression of a downstream gene was significantly lower in the transposon mutant than in the wild-type strain and the biofilm-enhanced deletion mutant. Transformation of this gene into the transposon mutant partially restored production of biofilm. Mice challenged by intravenous injection with the biofilm negative mutant strain showed significantly reduced numbers of colony-forming units in the blood, compared with mice challenged with the biofilm-enhanced deletion mutant and the wild-type. These results indicate that bop is involved in production of biofilm and probably regulates expression of biofilm in the E. faecalis strain tested. PMID- 14745700 TI - Susceptibility of common fibroblast cell lines to transmissible spongiform encephalopathy agents. AB - The risk of contamination of tissue culture cells with transmissible spongiform encephalopathy (TSE) agents as a result of the use of animal products as medium components has been considered to be low, in part, because only a few brain derived cell lines have been reported to be susceptible to TSE infection. In the present study, we demonstrate that the common laboratory fibroblast cell lines NIH/3T3 and L929, which express low levels of cellular mouse prion protein, are susceptible to infection with mouse-adapted scrapie. Our results show that the susceptibility of a cell line to TSE infection cannot be predicted on the basis of its tissue origin or its level of expression of the cellular prion protein, and they suggest that any cell line expressing normal host prion protein could have the potential to support propagation of TSE agents. Thus, testing of cells for TSE susceptibility might be necessary for all cell lines that are routinely used in vaccine production and in other medical applications. PMID- 14745701 TI - Management of influenza in households: a prospective, randomized comparison of oseltamivir treatment with or without postexposure prophylaxis. AB - We determined the efficacy of postexposure prophylaxis (PEP) and treatment of ill index cases with oseltamivir, in an attempt to prevent influenza transmission in households, in a study conducted in 277 households with 298 index cases (62% with laboratory-confirmed influenza) and 812 contacts aged > or =1 year. Contacts were randomized by household to receive treatment (5 days; n=402), if illness developed, or PEP for 10 days (n=410), and the number of households with at least 1 contact developing laboratory-confirmed influenza was measured. PEP provided a protective efficacy of 58.5% (95% confidence interval [CI], 15.6%-79.6%; P=.0114) for households against proven influenza and 68.0% (95% CI, 34.9%-84.2%; P=.0017) for individual contacts, compared with treatment of index cases alone. No oseltamivir-resistant variants were detected in treated index cases or contacts. PEP of household contacts of those with influenza reduces the secondary spread of influenza in families when the initial household case is treated. PMID- 14745702 TI - Development and validation of a clinical prediction rule for hospitalization due to pneumonia or influenza or death during influenza epidemics among community dwelling elderly persons. AB - BACKGROUND: Uncertainties among health care providers and patients about the risk of serious influenza-associated complications and the potential benefits of vaccination may contribute to unsatisfactorily low influenza vaccination rates. To quantify the risk of serious outcomes (hospitalization due to pneumonia or influenza or death due to any cause) during influenza seasons, we developed a clinical prediction rule for the probability of hospitalization due to pneumonia or influenza or death among elderly persons. METHODS: We developed the clinical prediction rule using data from linked administrative databases in a cohort of 16,280 noninstitutionalized and unvaccinated elderly persons. Validation of the rule was conducted in 5 unvaccinated and 6 vaccinated cohorts, each consisting of >11,000 elderly members of 3 managed care organizations. Logistic regression was used to produce a prognostic score on the basis of the following predictors: age; sex; presence of pulmonary, cardiac, and renal disease; dementia or stroke and cancer; number of outpatient visits; and hospitalization due to pneumonia or influenza during the previous year. RESULTS: Reliability of the regression model was good (P=.65, by goodness-of-fit test), and it discriminated well between those who did and those who did not experience an outcome (area under the receiver-operating curve, 0.83; 95% confidence interval, 0.81-0.85). Validation revealed moderately lower but acceptable discriminating values (0.72-0.81). In the derivation cohort, the prognostic accuracy of the rule was high when a cutoff score for the upper 50th percentile was used: > or =10 of 1000 subjects with a score in the upper 50th percentile were predicted to have an outcome, and 89% of all outcomes were observed in this high-risk group, whereas <10 of 1000 subjects with a score in the lower 50th percentile were predicted to have an outcome, and only 11% of outcomes occurred in this group. Among unvaccinated subjects in the single-derivation cohort and the 11 validation cohorts combined, the outcome event rates were 35 events/1000 subjects in the higher-risk group and 6 events/1000 subjects in the lower-risk group. With vaccination, these event rates dropped by 15 events/1000 subjects and 2 events/1000 subjects, respectively. CONCLUSIONS: This prediction rule may be a useful tool to complement other age based strategies, to further encourage vaccination, especially among those at the highest risk of serious complications due to influenza. PMID- 14745703 TI - Protection of mice with recombinant influenza virus neuraminidase. AB - Contemporary influenza vaccines are standardized with respect to their content of hemagglutinin, the major virus antigen. Although the immunizing effect of viral neuraminidase--the less abundant of the 2 major surface glycoproteins--has been well documented in experimental animals, the importance of the purified recombinant protein has not yet been adequately assessed in animals or humans. We demonstrate that different lots of a baculovirus-derived recombinant N2 protein, in the absence of other influenza virus proteins, can induce neuraminidase specific antibodies, reduce the replication of both homologous and heterovariant virus in mice, and suppress disease, as it is manifested by total body weight loss. PMID- 14745704 TI - Phase 2 evaluation of parainfluenza type 3 cold passage mutant 45 live attenuated vaccine in healthy children 6-18 months old. AB - A phase 2 evaluation of live attenuated parainfluenza type 3 (PIV3)-cold passage mutant 45 (cp45) vaccine was conducted in 380 children 6-18 months old; 226 children (59%) were seronegative for PIV3. Of the 226 seronegative children, 114 received PIV3-cp45 vaccine, and 112 received placebo. No significant difference in the occurrence of adverse events (i.e., runny nose, cough, or temperature > or =38 degrees C) was noted during the 14 days after vaccination. There was no difference between groups in the occurrence of acute otitis media or serous otitis media. Paired serum samples were available for 109 of the seronegative vaccine recipients and for 110 of the seronegative placebo recipients; 84% of seronegative vaccine recipients developed a > or =4-fold increase in antibody titers. The geometric mean antibody titer after vaccination was 1 : 25 in the vaccine group and <1 : 4 in the placebo group. PIV3-cp45 vaccine was safe and immunogenic in seronegative children and should be evaluated for efficacy in a phase 3 field trial. PMID- 14745705 TI - Two years' prospective collection of molecular and epidemiological data shows limited spread of hepatitis A virus outside risk groups in Amsterdam, 2000-2002. AB - We performed a viral sequencing study on samples representing all reported primary cases of acute hepatitis A virus (HAV) infection reported for 2 years in Amsterdam. Two regions of HAV RNA were amplified, sequenced, and used for phylogenetic analysis. Of 156 cases, strains of 104 isolates (66.6%) clustered into 3 genotypes: 1A, 1B, and 3. Two separate transmission circles occurred, without mutual interrelation. In genotype 1A, 4 clusters occurred in men having sex with men (MSM), and the fifth cluster was related to a virus from Morocco. In genotype 1B, 6 small clusters were directly related to the Moroccan virus. In genotype 3, strains were related to a virus from Pakistan. Our analysis indicates that, to stop transmission of HAV in Amsterdam, the entire MSM population and travelers to countries where HAV is endemic, especially children, should be vaccinated. Prevention strategies need not include the vaccination of all children living in Amsterdam. PMID- 14745706 TI - Biology of Epstein-Barr virus during infectious mononucleosis. AB - Infectious mononucleosis is the clinical manifestation of primary infection with Epstein-Barr virus (EBV). We monitored primary infection during convalescence and during the establishment of persistent infection. The profiles of EBV strains in the oral cavity and in peripheral blood were determined by use of a heteroduplex tracking assay specific for the EBV gene encoding latent membrane protein 1. Multiple EBV strains were detected in most patients and persisted in and were possibly transmitted among 3 distinct compartments of infection, including the oral cavity, peripheral blood lymphocytes, and the cell-free fraction of the blood plasma. We also tracked transmission of multiple strains from an asymptomatic carrier to a patient diagnosed with primary EBV infection. These data reveal that primary EBV infection is complex, with transmission of multiple strains and clear differences in relative abundance of strains in distinct compartments. PMID- 14745707 TI - Dose-dependent neutralizing-antibody responses to vaccinia. AB - To evaluate the humoral immune responses to smallpox-vaccine stocks currently available in the United States (Dryvax; Wyeth) and to generate data for comparison of responses to newly produced lots of smallpox vaccine, we evaluated dose-response effects, using undiluted and diluted smallpox vaccine. At 28 and 56 days after vaccination, serum samples were obtained from vaccinated subjects (N=674) who had participated in a randomized, single-blinded trial of an undiluted or a 1 : 5 or 1 : 10 dilution of smallpox vaccine and who subsequently were tested for plaque-reduction neutralizing-antibody titer. All subjects who developed a vesicle after vaccination also developed neutralizing antibodies by day 28. Subjects given either a 1 : 5 or 1 : 10 dilution of vaccinia had significantly higher neutralizing-antibody titers than did subjects given undiluted vaccine. Larger lesion size and fever after vaccination were associated with significantly higher neutralizing-antibody titers after vaccination. PMID- 14745708 TI - No differences in cellular immune responses between asymptomatic HIV type 1- and type 2-infected Gambian patients. AB - Fewer people infected with human immunodeficiency virus (HIV) type 2 progress to acquired immunodeficiency syndrome, compared with those infected with HIV-1. To understand the immune mechanisms leading to slow progression in HIV-2 infection, cell-mediated immune responses were compared between the 2 infections in asymptomatic subjects with a CD4 cell count > or =20%. Interferon- gamma release from T lymphocytes and the cytotoxicity of CD8+ T lymphocytes were measured by ELISPOT and 51Cr release assays. The level of responses and the proportion of responders were similar in the 2 infections, despite a 20-fold difference in their geometric mean plasma virus loads. The proliferation of CD4+ T helper cells, which was evaluated by thymidine incorporation, was not different between the 2 infections. Contrary to widely held views, our results suggest that nonprogression in HIV-2 infection may not be due to more vigorous immune responses. PMID- 14745709 TI - CXCR4-tropic HIV-1 suppresses replication of CCR5-tropic HIV-1 in human lymphoid tissue by selective induction of CC-chemokines. AB - In infected individuals, human immunodeficiency virus type 1 (HIV-1) exist as a "swarm" of quasi species compartmentalized in tissues where individual viral variants may interact locally. We have used human lymphoid tissue, where the critical events of HIV disease occur, to study local interactions in model HIV-1 binary swarms ex vivo. We infected tissue blocks with binary mixtures consisting either of CCR5-dependent and CXCR4-dependent variants or of 2 dual-tropic HIV-1 variants, of which one is skewed to utilization of CXCR4 and the other of CCR5. HIV-1 variants that use CXCR4 suppress replication of CCR5-dependent HIV-1 variants, whereas CCR5-dependent HIV-1 variants do not affect replication of CXCR4-dependent HIV-1. CC-chemokines that inhibit replication of CCR5-dependent HIV-1 variants were up-regulated by CXCR4-dependent HIV-1, thus possibly contributing to this suppression. Tissue-specific chemokine/cytokine network modulations triggered by individual HIV-1 variants may be an important mechanism of local interactions among HIV-1 quasi species in infected tissue. PMID- 14745710 TI - Association of T cell proliferative responses and phenotype with virus control in chronic progressive HIV-1 disease. AB - The role of T cell immunity in virus control during chronic infection was examined in 79 human immunodeficiency virus type 1 (HIV-1)-infected subjects randomized to receive antiretroviral therapy. HIV-1 p24-specific responses were detected in 20% of the subjects at baseline, increasing to 28% of the subjects at weeks 16-24. Induction of virologic suppression was associated with lower plasma HIV-1 RNA levels and a higher percentage of Fas+ CD8+ T cells at baseline, whereas maintenance of suppression was associated with higher CD4+ T cell counts and, marginally, with a higher percentage of Fas+ CD4+ T cells at weeks 16-24. These findings indicate that Fas coexpression on T cells, in addition to plasma HIV-1 RNA levels and CD4+ T cell counts, may predict virologic outcome. PMID- 14745711 TI - Influence of human leukocyte antigen class II alleles on susceptibility to Entamoeba histolytica infection in Bangladeshi children. AB - BACKGROUND: The association of antibody responses with both innate and acquired immunity to amebiasis indicate that CD4+ T cells play a role in protection against Entamoeba histolytica infection. To test this hypothesis, we compared the genotype frequencies of human leukocyte antigen (HLA) class II alleles in a cohort of Bangladeshi children intensively monitored for E. histolytica infection for a 3-year period. METHODS: Using logistic regression, we calculated the odds of disease by genotype and by haplotype. RESULTS: The DQB1*0601 heterozygous and homozygous genotypes were found in 55% of E. histolytica-negative children but in only 34% of E. histolytica-positive children (overall odds ratio, 2.39; 95% confidence interval [CI], 1.26-4.54). Children who were heterozygous for the DQB1*0601/DRB1*1501 haplotype were 10.1 times (95% CI, 2.02-50.6) more likely to be both E. histolytica negative and serum anti-lectin immunoglobulin G negative at baseline. Other DQB1 and DRB1 alleles (DQB1*0202, DQB1*0301, and DRB1*0701) were not associated with any of the clinical outcomes related to amebiasis. CONCLUSION: A potential protective association was observed with the HLA class II allele DQB1*0601 and the heterozygous haplotype DQB1*0601/DRB1*1501. This association may explain why amebiasis does not occur in some children who are exposed to the parasite and implicates HLA class II-restricted immune responses in protection against E. histolytica infection. PMID- 14745712 TI - P75 tumor necrosis factor-receptor shedding occurs as a protective host response during African trypanosomiasis. AB - In experimental murine trypanosomiasis, resistance is often scored as the capacity to control peak parasitemia levels, which results in prolonged survival. Infection-induced pathology has not systematically been used as a resistance criterion. Because this parameter could be the most relevant for comparative analysis of natural and experimental infections, as well as for understanding of pathology-associated immune alterations, we analyzed Trypanosoma brucei infections in 4 different established conventional mouse models, as well as in tumor necrosis factor (TNF)-deficient and TNF-receptor-deficient mice. Results indicate the following: (1) there is no correlation between peak parasitemia control or survival and the induction of infection-associated anemia, loss of body weight, liver pathology, reduced locomotor activity, and general morbidity; (2) serum levels of TNF, interferon- gamma, and interleukin-10, which are known to affect survival, do not correlate with induction of pathology; and (3) infection-induced occurrence of lipopolysaccharide hypersensitivity does not correlate with survival. However, one parameter that was found to correlate with the inhibition of trypanosomiasis-associated pathology in all models was the shedding of soluble p75 TNF-receptor during peak parasitemia stages. These results are important for future cytokine and trypanosomiasis pathology studies, because the interplay between TNF and the soluble receptors it sheds has not been considered in either human clinical sleeping sickness studies or in veterinary trypanosomiasis research. PMID- 14745714 TI - Bacteria and diseases of unknown cause: hemolytic-uremic syndrome. PMID- 14745713 TI - Antibodies to variant surface antigens of Plasmodium falciparum-infected erythrocytes and adhesion inhibitory antibodies are associated with placental malaria and have overlapping and distinct targets. AB - We measured antibodies to chondroitin sulfate A (CSA)-binding and placental Plasmodium falciparum-infected red blood cells (PRBCs) among pregnant women with or without placental malaria. Immunoglobulin G to PRBC surface antigens was rare in uninfected primigravidae (3.7%), more prevalent in infected primigravidae (70%; P<.001), and common in infected (77%) and uninfected (83%) multigravidae. Similar patterns were seen for agglutinating antibodies, and antibodies were similar among women with past or active placental infection. PRBC adhesion to CSA was inhibited 60% by serum from infected primigravidae but 24% by serum from uninfected primigravidae (P=.025), whereas infection did not alter adhesion inhibition by multigravidae (77% inhibition)[corrected]. There was substantial heterogeneity in antibody type and levels. Antibodies did not correlate with parasite density or pregnancy outcome. Comparisons between antibodies suggest that adhesion-inhibitory antibodies and those to PRBC variant antigens have distinct and overlapping epitopes, may be acquired independently, and have different roles in immunity. PMID- 14745715 TI - Influenza virus and febrile convulsions. PMID- 14745717 TI - New technologies in research and diagnosis: implications and opportunities for Human Pathology, its authors, and its readership. PMID- 14745718 TI - Prognostic significance of thyroid transcription factor-1 expression in both early-stage conventional adenocarcinoma and bronchioloalveolar carcinoma of the lung. AB - Thyroid transcription factor 1 (TTF-1) is a diagnostic immunohistochemical marker for primary pulmonary neoplasms, but its utility as a prognostic marker is not well established. Surgical specimens from 100 cases of pulmonary adenocarcinoma were retrieved from the hospital computer system, including 50 cases of conventional adenocarcinoma (CA) and 50 cases of bronchioloalveolar adenocarcinoma (BAC) (32 nonmucinous type and 18 mucinous type). Representative sections were immunostained for TTF-1. Positive immunohistochemical study was correlated with other prognostic parameters. In the CA group, strong or moderate TTF-1 expression was seen in 30 of 50 (60%) patients and was associated with significantly better survival compared with those patients having weak staining (7 cases; 14%) or negative staining (13 cases; 26%) (P <0.01; log-rank test). Spearman and Pearson's correlation showed no significant correlation between TTF 1 expression and tumor grade, size, recurrence, or vascular invasion; therefore, TTF-1 was considered an independent predictor of survival. In the BAC group, TTF 1 was strongly expressed in 34 of 50 cases (68%) and was negative in 16 of 50 cases (32%), including 14 mucinous BACs. Although TTF-1 immunoreactivity was not statistically associated with good survival in BAC patients, those patients with strong immunohistochemical expression showed a trend toward longer survival. Our results indicate that TTF-1 positivity is an independent predictor of better survival, especially in patients with CA. Mucinous and nonmucinous BACs exhibit disparate staining patterns with TTF-1, with nonmucinous BAC demonstrating greater positivity. Although nonmucinous BAC patients showing strong positive staining had longer survival, the difference was not statistically significant, which is probably related to the overall good survival of patients with early stage BAC. PMID- 14745719 TI - Carcinoma of lung with rhabdoid features. AB - Lung tumors with rhabdoid features, included as variants of large cell carcinoma in the 1999 World Health Organization classification of lung tumors, are rare and have an aggressive clinical course. We report 11 patients with primary lung tumors with rhabdoid features and review the literature on this uncommon tumor. We examined samples from 7 primary (6 resections, 1 biopsy) and 4 metastatic tumor samples. All specimens were stained with immunohistochemical stains for pancytokeratin (CK), cytokeratin 7 (CK7), cytokeratin (CK20), thyroid transcription factor-1 (TTF-1), and vimentin. The patients were 7 men and 4 women whose ages ranged from 35 to 70 years. Nine patients presented with respiratory symptoms, and 9 patients had a history of heavy smoking. One patient had TNM stage I tumor, 3 had stage III tumors, and 6 had stage IV tumors at presentation; tumor stage could not be determined in 1 patient. Histological examination of these tumors showed typical rhabdoid cells: large cells with abundant cytoplasm, a large eccentric nucleus with a central macronucleolus, and a rounded eosinophilic cytoplasmic inclusion that sometimes caused nuclear indentation. These cells constituted 10% to 90% of the tumor. The "parent" neoplasm was sarcomatoid carcinoma and adenocarcinoma in 4 cases each and was large cell undifferentiated carcinoma in 3 cases. Cytoplasmic staining in the rhabdoid cells was seen in 9 of 11 cases for CK, in 4 of 10 cases for CK7, and in all 11 cases for vimentin. Nuclear staining for TTF-1 in the rhabdoid cells was absent in all 11 cases, and cytoplasmic staining for CK20 was negative in the rhabdoid cells in all 10 cases studied. Of the 9 patients with available follow-up information, 8 died of disease, and 1 is alive with no evidence of disease 20 months after the initial diagnosis. We conclude that rhabdoid features can occur in a variety of lung tumors, including sarcomatoid carcinoma. Recognizing these lesions is important because of their possibly aggressive clinical course. PMID- 14745720 TI - Cutaneous myoepithelioma: a clinicopathologic and immunohistochemical study of 14 cases. AB - Analogous to mixed tumors of salivary glands (" pleomorphic adenomas" ), cutaneous mixed tumors (" chondroid syringomas" ) contain a ductal (epithelial) component and a variably prominent myoepithelial component. Tumors showing purely myoepithelial differentiation (myoepitheliomas) have only recently been recognized to arise in the dermis, and to date very few cases have been described. To characterize these tumors further, 14 cutaneous myoepithelial tumors were retrieved from the authors' consult files. Eleven patients were male and 3 were female; their median age was 22.5 years (range, 10 to 63 years), and 7 patients were between 10 and 20 years old. Tumor size ranged from 0.5 to 2.5 cm (mean, 1.1 cm). Most tumors arose on the extremities: 6 on the upper limbs, 6 on the lower limbs, and 1 each on the back and nose. Ten tumors were limited to the dermis, and 5 also extended into superficial subcutis. Thirteen tumors were myoepitheliomas (lacking ductal differentiation), and 1 tumor was a myoepithelial carcinoma (exhibiting severe cytological atypia and a high mitotic rate). Histologically, 7 tumors were solid, composed of ovoid to spindled, histiocytoid, or epithelioid cells with no significant stroma, and 7 were predominantly lobulated, with cords or nests of epithelioid, plasmacytoid, or spindled cells with a variably reticular architecture and a chondromyxoid or collagenous/hyalinized stroma. One tumor was composed solely of plasmacytoid (hyaline) cells, and 1 exhibited extensive adipocytic differentiation. Among the 13 myoepitheliomas, mitoses ranged from 0 to 6 per 10 high-power fields (HPFs) (mean, 1.5); 8 tumors contained no mitoses. The myoepithelial carcinoma had 39 mitoses per 10 HPFs. By immunohistochemistry, all cases were reactive for epithelial markers (keratins and/or epithelial membrane antigen [EMA]); 13 of 14 (93%) expressed S-100 protein, 10 of 11 expressed (91%) calponin, 11 of 14 (79%) expressed EMA, 9 of 14 (64%) expressed keratins, 8 of 14 (57%) expressed smooth muscle actin, 7 of 14 (50%) expressed glial fibrillary acidic protein, 3 of 11 (27%) expressed p63, and 1 of 6 (17%) expressed desmin. All 5 cases without keratin staining were diffusely positive for EMA, and all of these cases showed a solid growth pattern. Follow-up was available for 8 patients (median follow-up, 40 months; range, 6 months to 9 years); 3 tumors (38%) recurred locally, and 1 tumor (13%) also metastasized to the lymph nodes. The case that resulted in recurrence and metastasis had the highest mitotic rate (6 per 10 HPFs) of the cytologically benign tumors. Follow-up information was not available for the myoepithelial carcinoma. This study suggests that approximately 50% of cutaneous myoepitheliomas are distinctive lesions composed of a solid proliferation of cells with abundant eosinophilic syncytial cytoplasm, which often lack immunostaining for keratin, whereas the remainder demonstrate focally reticular architecture and myxoid stroma or plasmacytoid cells, similar to their counterparts in salivary gland and soft tissue. Whereas most cutaneous myoepitheliomas behave in a benign fashion, there is apparently a significant risk for local recurrence but a low metastatic potential. PMID- 14745721 TI - Expression and localization of mutant p16 proteins in melanocytic lesions from familial melanoma patients. AB - Little is known about the correlation between the loss of p16 expression and tumor progression in familial melanoma; no systematic study has been conducted on p16 expression in melanocytic tumors from patients carrying germline CDKN2A mutations. We analyzed 98 early primary lesions from familial patients, previously tested for germline CDKN2A status, by quantitative immunohistochemistry using 3 p16 antibodies. We found that p16 expression was inversely correlated with tumor progression and was significantly lower in melanomas, including in situ lesions, than in nevi. Of other features analyzed, tumor thickness showed the most significant correlation with p16 levels. Lesions from mutation-negative patients displayed combined nuclear and cytoplasmic staining. However, some mutation-positive lesions (ie, G101W, 113insR, M53I, R24P, and 33ins24), including benign nevi, showed nuclear mislocalization, confirming previous studies suggesting that subcellular distribution indicates functional impairment of p16. PMID- 14745722 TI - Normal and pathological human testes express hormone-sensitive lipase and the lipid receptors CLA-1/SR-BI and CD36. AB - Numerous studies have demonstrated the important role of cholesterol and cholesteryl esters in tumor cell proliferation and progression of cancer. However, few studies have focused on the role of lipid transporters and lipases in cancer development and progression. The present study examined the expression of hormone-sensitive lipase (HSL) and the scavenger receptors CLA-1/SR-BI and CD36 in normal human testis and in nontumor and tumor testicular disorders by immunohistochemistry and Western blotting analysis. In normal young testes, immunoreaction to CLA-1/SR-BI was found in the spermatid acrosomic vesicle and on the surface of Sertoli and Leydig cells. HSL was detected in spermatogonia, the Golgi region of spermatocytes, the nucleus of spermatids, and the cytoplasm of both Sertoli and Leydig cells. Elderly testes and testes with hypospermatogenesis showed a similar staining pattern to that of normal young testes except for CD36, which was expressed in Sertoli cells. Cryptorchid testes demonstrated intense labeling to HSL and weak labeling to SR-BI in Sertoli cells (nucleus and cytoplasm) and Leydig cells (cytoplasm). Seminiferous tubules with intratubular germ cell neoplasia exhibited intense immunolabeling to the 3 lipid receptors in the surface of neoplastic cells and to HSL in the nucleus. In seminoma and spermatocytic seminoma, neoplastic cells labeled to HSL but failed to stain with antilipid receptors; in the seminiferous tubules at the periphery of the tumour, Charcot-Bottcher crystalloids of Sertoli cells were strongly positive to CLA-1. Testes with mature teratoma showed a weak reaction to CD36 and SR-BI in some cells of enteric-type glands, and immature teratoma were exclusively immunolabeled with HSL. Western blotting analysis revealed that multiple bands were immunolabeled, with differences seen between normal and pathological testes. The results of this study indicate that the presence of lipid receptors (CLA-1/SR BI) and hormone-sensitive lipase in Leydig cells suggests a role of these proteins in steroidogenesis. Also, these proteins seem to be involved in spermiogenesis, as their labeling in spermatids suggests. In nonmalignant and malignant pathologies, cholesterol metabolism is probably altered, and HSL labeling in neoplastic germ cell nuclei suggests a still-unknown function of this enzyme, probably related to cell cycle regulation. PMID- 14745723 TI - Negative 34betaE12 staining in a small focus of atypical glands on prostate needle biopsy: a follow-up study of 332 cases. AB - Atypical glands on prostate needle biopsy with a negative 34betaE12 (cytokeratin 903; CK903) immunostain, indicating a lack of a basal cell layer, are typically diagnostic of prostate cancer. However, in certain cases a negative 34betaE12 immunostain in a small focus of atypical glands is still not convincing enough to make the diagnosis of cancer. This study is the first report to evaluate the incidence of prostate cancer on follow-up biopsy in individuals with this diagnosis. A total of 543 men who had prostate core biopsy specimens diagnosed as a small focus of atypical-appearing glands with a negative 34betaE12 immunostain between January 1, 1997 and December 31, 2000 were selected for study. Some 61% of these 543 individuals (n = 332) had undergone at least one follow-up biopsy procedure. Of these, 43% of repeat biopsy cases (n = 142) were diagnostic of prostate cancer. A total of 46 individuals had at least 2 follow-up biopsy procedures, with 48% of these (n = 22) being diagnosed as cancer. The Gleason grades of the detected carcinomas were broken down as follows: Gleason grade 3 + 2 = 5, 6%; grade 3 + 3 = 6, 86%; grade 3 + 4 = 7, 1%; grade 4 + 3 = 7, 4%; and grade 4 + 4 = 8, 3%. The median amount of time to the first follow-up biopsy was 79 days, with 52% of follow-up biopsies performed within 90 days. A negative 34betaE12 immunohistochemical stain in a small focus of atypical glands is not associated with an increased prediction of prostate cancer on follow-up biopsy (43%), compared with previously published data for "small focus of atypical glands" alone (approximately 45%). Because 48% of men with an initial negative biopsy and multiple follow-up biopsy procedures were found to have cancer, more than one repeat biopsy session or more extensive sampling on the first repeat biopsy procedure may be necessary to maximize the identification of cancer. This finding is similar to that found in men with atypical diagnoses in general, without a negative 34betaE12 immunohistochemical stain. Only half of all individuals with a diagnosis of 34betaE12-negative focus of atypical glands underwent repeat biopsy within 3 months. Urologists need to be educated as to the significance of an atypical diagnosis and the need for repeat biopsy. In a small focus of atypical glands on prostate biopsy, negative staining for 34betaE12 should not necessarily lead to a definitive malignant diagnosis in all cases, because almost half of these biopsies on follow-up sampling are benign. PMID- 14745724 TI - Immunohistochemical distribution of Toll-like receptor 4 in term and preterm human placentas from normal and complicated pregnancy including chorioamnionitis. AB - Recently discovered Toll-like receptors (TLRs) are essential for the induction of innate immune responses. One member of the TLR family, TLR4 mediates lipopolysaccharide (bacterial endotoxin)-induced inflammatory responses. Although the innate immune system appears to be important in the pathogenesis of infection induced preterm delivery, the distribution of TLRs in human placenta is poorly understood. Here we investigated the expression of TLR4 protein in 43 human placentas obtained from normal and complicated pregnancies delivered in the second and third trimesters, using immunohistochemistry. TLR4 was localized to the extravillous trophoblasts, intermediate trophoblasts/X cells in the degenerative villi, and villous Hofbauer cells of all preterm and term placentas examined and to the inflammatory cells in placentas with chorioamnionitis (CAM). The villous Hofbauer cells of preterm CAM placentas demonstrated increased TLR4 immunoreactivity compared with those of preterm placentas without CAM or those of term placentas with or without CAM. These results suggest an important role of the villous Hofbauer cells in the activation of innate immune system in response to infectious pathogens in preterm placentas. Elucidation of biological functions of placental TLR4 under physiological conditions requires further investigation. PMID- 14745725 TI - Expression of the mitogen-inducible gene-2 (mig-2) is elevated in human uterine leiomyomas but not in leiomyosarcomas. AB - To investigate the role of a recently cloned cell proliferation-related gene, mig 2 (mitogen-inducible gene-2), in the growth of uterine leiomyomas, this gene's expression at mRNA and protein levels was examined in normal myometrium, leiomyomas, and leiomyosarcomas of the uterus. Northern blotting, Western blotting, and immunohistochemical staining demonstrated that mig-2 expression was increased in leiomyomas compared with normal myometrium, especially during the secretory phase of the menstrual cycle. In contrast, the mig-2 expression was greatly decreased in leiomyosarcomas. Direct sequencing of the whole coding region of mig-2 cDNA and Southern blotting revealed that mig-2 alterations, such as mutations, rearrangement, and amplification, were not present in either leiomyomas or leiomyosarcomas. These findings suggest that mig-2 expression is transcriptionally elevated in leiomyomas and could be involved in its hormone mediated growth of leiomyomas of the uterus. PMID- 14745726 TI - Decreased expression of transforming growth factor-beta II receptor is associated with that of p27KIP1 in giant cell tumor of bone: a possible link between transforming growth factor-beta and cell cycle-related protein. AB - Transforming growth factor (TGF)-beta is a potential regulator of cell growth that sends its signals through a heteromeric complex composed of type I and II receptors (IR and IIR). This study examined a correlation between TGF-beta1, TGF beta-IR and -IIR, and cell cycle-related proteins in giant cell tumor (GCT) of bone, using immunohistochemical and Western blot analysis. First, an immunohistochemical study for TGF-beta1, TGF-beta-IR and -IIR, p27KIP1 (p27), p21WAF1 (p21), cyclin D1, and cyclin E was carried out on 92 cases of GCT of bone; the expression of these proteins was evaluated in multinucleated giant cells (MGCs) and mononucleated stromal cells (MSCs) separately; and proliferative activity was assessed using MIB-1. Next, to confirm our immunohistochemical results, Western blot analysis was performed in 19 cases for which frozen samples were available. Immunoreactivity for TGF-beta-IR and -IIR showed a tendency to be greater in MGCs than in MSCs; however, no differences were observed in TGF-beta1. Cyclin D1 expression was correlated with the occurrence of vascular invasion in both MGCs and MSCs (P = 0.0255 and 0.0183, respectively). The expression of TGF beta-IIR and p27 was concordantly decreased in both MGSs and MSCs (P = 0.0014 and 0.0317, respectively). The expression for TGF-beta-IIR and p27 in Western blot analysis was related to the results from immunohistochemical analysis, and the expression of TGF-beta-IIR and p27 was concordant in almost all GCT cases. Furthermore, there was a statistically significant inverse relationship between p27 expression and MIB-1 labeling index in MSCs (P = 0.0397). In GCT of bone, TGF beta-IIR and p27 expression were concordantly decreased; this result supports the possibility that these 2 factors may play an important role in cell proliferation of this tumor. Furthermore, our results provide a possible link between the effects of extracellular growth factors and cell cycle control. In addition, p27 expression may be a useful indicator of cell proliferation in MSCs of this tumor. PMID- 14745727 TI - Osteofibrous dysplasia and adamantinoma: correlation of proto-oncogene product and matrix protein expression. AB - To investigate the relationship between osteofibrous dysplasia (OFD) and adamantinoma, we analyzed the expression of several proto-oncogene products and extracellular matrix proteins by immunohistochemistry and correlated our results with histological and ultrastructural findings. C-fos and c-jun, but not c-Met, were observed in OFD and in the fibrous and epithelial components of differentiated and classical adamantinomas. Staining for collagen IV, laminin and galectin-3, a laminin binding protein was seen in OFD and around cell nests in adamantinoma. E-, P-, and N-cadherin expression was found in all cases of classical adamantinoma, but not in differentiated adamantinoma or OFD. Osteonectin was detected in both the epithelial and fibrous components of adamantinomas, but osteopontin and osteocalcin were not seen in classical adamantinomas. The results show common expression of a number of oncoproteins and bone matrix proteins in adamantinoma and OFD, some of which are associated with mesenchymal-to-epithelial cell transformation. These findings would be in keeping with the hypothesis that OFD represents a precursor lesion of adamantinoma. Differential expression of a number of bone matrix protein in adamantinoma may also be of diagnostic use in distinguishing these 2 lesions immunohistochemically. PMID- 14745728 TI - Altered plakoglobin expression at mRNA and protein levels correlates with clinical outcome in patients with oropharynx squamous carcinomas. AB - Previous studies have established that expression of plakoglobin is down regulated during malignant transformation. The aim of this study was to evaluate for the first time the expression of plakoglobin at the mRNA and protein levels in primary oropharyngeal squamous cell carcinomas (SCCs) and determine the extent to which the patterns of expression correlated with clinical parameters. Plakoglobin expression was evaluated in 37 new tumor cases and normal oral epithelium using immunofluorescence, reverse transcriptase-polymerase chain reaction (RT-PCR), and Northern and Western blotting analysis. The results indicated that the steady-state levels of plakoglobin protein were down-regulated in all tumors compared with normal epithelium. Furthermore, in 87.1% of the tumors, plakoglobin immunoreactivity displayed an abnormal cytoplasmic localization that was inversely correlated with tumor size and directly correlated with a poor clinical outcome for the patient. Northern blotting analysis revealed that down-regulation of mRNA expression occurred in only 65.6% of the tumors, with plakoglobin mRNA levels similar to normal epithelium in the remaining cases. In the tumors expressing mRNA levels similar to those of normal tissue, a 3.7-kb transcript was detected in addition to the expected 3.4-kb transcript observed in normal epithelium. RT-PCR analysis of the 3' untranslated region of the 3.7-kb plakoglobin mRNA transcript identified a 297-base insertion from +2369 to +2666 that had been previously reported only in transformed cell lines (GenBank M23410). Interestingly, the prognosis was poor for patients with tumors expressing both RNA transcripts. These results are consistent with the concept that complex regulation of plakoglobin expression and intracellular routing may contribute to malignant transformation. The study also shows evidence that the level of expression and intracellular localization of plakoglobin may be useful in predicting the course of disease in patients with oropharyngeal SCC. PMID- 14745729 TI - P53, N- and K-Ras, and beta-catenin gene mutations and prognostic factors in nasal NK/T-cell lymphoma from Hokkaido, Japan. AB - We have shown previously that nasal natural killer (NK)/T-cell lymphoma was associated with Epstein-Barr virus (EBV) and had peculiar clinical features. However, little is known about its biological and genetic changes. The aim of this study is to determine the p53, N- and K-ras, and beta-catenin status in this lymphoma in relation to EBV status and clinical features. The study group consisted of 32 Japanese patients with nasal NK/T-cell lymphoma. The p53 and beta catenin expression, phenotype, and EBV-oncogenic protein latent membrane protein type 1 (LMP-1) were determined by immunoperoxidase staining. The presence of EBV encoded small nuclear early region (EBER) RNA was determined by in situ hybridization. The p53 mutations (exons 5 to 9), N- and K-ras mutations (exons 1 and 2), and beta-catenin mutations (exon 3) were analyzed by direct sequencing of the PCR-amplified products that were obtained from laser-microdissected tissues. CD56, CD43, and CD3 were expressed in 32 (100%), in 31 (96%), and in 18 (56%) tumors, respectively. EBER RNA was detected in 31 (96%) tumors. LMP-1 was expressed in 15 (48%) tumors, and p53 and beta-catenin protein were overexpressed in 18 (56%) and 4 (13%) tumors, respectively. Six mutations of the p53 gene, 1 mutation of each N- and K-ras gene, and 8 mutations of beta-catenin gene were detected in 6 (19%), 1 (3%), and 5 (16%) tumors, respectively. The p53 missense mutation was associated with LMP-1 expression (P = 0.038), but not with p53 overexpression. Kaplan-Meier analysis as well as univariate analysis using Cox proportional hazards model showed that high lactate dehydrogenase (LDH) level (P = 0.009, P = 0.0100, respectively), large cell, immunoblastoid polymorphous histology (P = 0.005, P = 0.0162, respectively), and p53 missense mutations (P = 0.021, P = 0.0342, respectively) were significantly related to worse cause specific survival. Multivariate analysis showed that p53 missense mutation was the most independent among these 3 factors. Although the incidence of thep53, N- and K-ras, and beta-catenin gene mutations is not high, p53 missense mutation has a prognostic value for aggressive course in nasal NK/T-cell lymphoma. PMID- 14745730 TI - Histological score for cells with irregular nuclear contours for the diagnosis of reflux esophagitis in children. AB - Histological criteria for the diagnosis of reflux esophagitis include basal zone hyperplasia, stromal papillae elongation, and inflammatory infiltrate. However, endoscopic esophageal biopsy specimens may include little or no lamina propria. Intraepithelial T lymphocytes, seen in hematoxylin and eosin-stained sections as cells with irregular nuclear contours (CINC), may have a higher density in children with esophagitis. We evaluated the diagnostic accuracy of a numerical score built up by grading the "classical" parameters and its correlation with CINC density in grasp biopsy specimens obtained from children undergoing esophagogastroduodenoscopy with and without esophagitis. We analyzed esophageal biopsy specimens from 349 children (median age, 5 years) subdivided in 4 groups according to the previous routine histology report: group 1, 144 children with esophagitis; group 2, 65 controls; group 3, 51 children with dubious esophagitis; and group 4, 75 children with esophagitis on endoscopy but a normal histology report. A numerical value was assigned to each parameter; the sum of these values represented the histological score. We also evaluated intraepithelial CINC density (ie, number of CINC per high-power field). We separately analyzed histological sections with and without lamina propria. For both total score and for CINC density, we calculated a cutoff using a receiver operating characteristic curve. Cutoffs of 6 for score and of 4 for CINC density provided the best sensitivity and specificity. Sensitivity of the histological score was better in biopsy specimens containing lamina propria (94%) than in those without lamina propria (4%). Sensitivity of CINC density was satisfactory in both specimens with (78%) and without (75%) lamina propria. Specificity was satisfactory for both parameters. In conclusion, when lamina propria was present in sections of endoscopic esophageal biopsy specimens, histological score provided a better diagnostic accuracy for the diagnosis of esophagitis. However, when no lamina propria was present, as was the case in 67% of our children, CINC density had better sensitivity. In addition, this latter parameter showed esophageal mucosa damage in 34% of previously dubious cases or cases with esophagitis at endoscopy but a previous routine histology report of normal mucosa. PMID- 14745731 TI - Infrequency of microsatellite instability in complete and incomplete gastric intestinal metaplasia. AB - Chronic inflammation may be associated with microsatellite instability (MSI). To test the hypothesis that MSI frequently occurs in gastric intestinal metaplasia, we examined gastric biopsies from 58 subjects from an area of high risk for gastric cancer. These were selected to have 2 types of intestinal metaplasia: complete (31 subjects) and incomplete or mixed-type (27 subjects). None of the subjects had gastric cancer, but 95% had chronic inflammation with Helicobacter pylori. We used laser capture microdissection to retrieve metaplastic glands to compare with lymphocytes microdissected from the adjacent gastric mucosae in the same subjects. We performed microsatellite analysis using 6 microsatellite loci, including BAT26. None of the cases were found to have reproducible MSI, and only 1 case showed loss of heterozygosity at 1 marker, D3S1067. To test the sensitivity of our assay, we mixed templates to produce bands of different mobility and found that we could detect an aberrant microsatellite pattern if only 2% of the DNA showed that pattern. Our results indicate that MSI is a rare event in intestinal metaplasia in subjects who do not have gastric cancer. PMID- 14745732 TI - Reversibility of hepatitis C virus-related cirrhosis. AB - The aim of this retrospective study was to determine the potential reversibility of hepatitis C virus (HCV) cirrhosis with the combined antifibrotic effects of interferon-alpha and the increasing frequency of sustained virologic response. Sixty-four HCV-cirrhotic immunocompetent patients who underwent antiviral therapies (interferon-alpha with or without ribavirin) and pretreatment and posttreatment liver biopsies were included (group 1). Resolution of cirrhosis was defined as a decrease in the fibrosis score from 4 to 2 or less by the Metavir score after blinded analysis by 2 independent pathologists. An additional group of 4 HCV-infected dialysis patients (group 2) who had received antiviral treatment, among whom 3 underwent a combined renal and liver transplantation allowing the analysis of the whole liver, was also studied. In 5 (all stage Child A) of the 64 cirrhotic patients (7.8%), the final biopsy showed only F2 to portal and periportal fibrosis with rare fibrous septa without nodule formation. Four of these 5 were complete sustained responders (negative PCR and normal ALT), and 1 was a relapser. In group 2, reversibility of cirrhosis was observed in 3 of the 4 patients and was clearly shown in 2 patients by the analysis of the whole-liver examination at the time of the hepatectomy preceding the transplantation. In conclusion, long-lasting suppression of the necroinflammatory activity of liver disease and/or antifibrogenetic effects of interferon-alpha may allow regression of cirrhosis. PMID- 14745733 TI - Age-dependent development of the splenic marginal zone in human infants is associated with different causes of death. AB - Infants are more susceptible to infections caused by T cell- independent type 2 (TI-2) polysaccharide antigens of certain encapsulated bacteria. Immune responses against this type of antigen are related to the splenic marginal zone (MZ). However, only few data exist on the age-dependent developmental stages of the human spleen in early childhood and on their association with different diseases. Therefore, the present study aimed to investigate spleens of a large number of children at very young ages (12 days to 32 months), derived from autopsy cases. Immunohistochemical labeling was performed on paraffin sections of 34 spleens using a panel of monoclonal antibodies. The shape and size of the white pulp compartments were examined and correlated to the cause of death of the children. Results show that the development of the different compartments was statistically age-dependent, but no clear-cut time point for the maturity of each compartment was seen. Furthermore, the MZ was significantly more often missing when sudden infant death (SID) and/or infection were the cause of death, compared with other violent or traumatic reasons that served as controls. This association supports the concept that an immature state of the spleen and especially of the MZ might contribute to the increased susceptibility to bacterial infections in young infants. PMID- 14745734 TI - Frequent EpCam protein expression in human carcinomas. AB - Expression of the transmembrane glycoprotein EpCam (epithelial cellular adhesion molecule) occurs in normal epithelium of different organs and was described in carcinomas of various sites. Specific anti-EpCam therapies are now being used in clinical trials. Thus, it is of interest to know which tumor types express or overexpress this protein, and in what frequency. We therefore analyzed EpCam expression by immunohistochemistry on a tissue microarray containing 3900 tissue samples of 134 different histological tumor types and subtypes. EpCam expression was detected in 98 of 131 tumor categories. At least a weak EpCam expression in >10% of tumors was observed in 87 of 131 different tumor categories. Adenocarcinomas of the colon (81%) and pancreas (78%), as well as hormone refractory adenocarcinomas of the prostate (71%), were identified as particularly promising therapy targets with a high fraction of strongly positive tumors. Most soft-tissue tumors and all lymphomas were EpCam negative. It is concluded that anti-EpCam therapies, if proven to be successful, will have broad applications in a wide variety of carcinomas. PMID- 14745735 TI - Diagnostic accuracy of second-opinion diagnoses based on still images. AB - Second opinion of histological specimens is an important part of the daily routine in anatomic pathology practices. Today, extramural second opinion can be easily obtained by sending still images via an electronic network. The aim of this study was to examine the diagnostic accuracy of second opinion diagnosis based on still images selected from glass slides of 90 archived cases originally referred for extramural second opinion. Two pathologists together diagnosed first the still images (phase 1) and then the glass slides (phase 2). Phase 1 and phase 2 diagnoses were compared with the original second opinion diagnoses (OSODs). The pathologists achieved the same diagnostic results in phase 1 and in phase 2 measured against the OSOD, 67.8% (n = 61) and 68.9% (n = 62) complete agreement, respectively. In 29 cases in phase 1, the diagnoses were discordant with the OSOD. Three cases had incorrect benign diagnoses and 8 cases had incorrect malignant diagnoses. There were 8 false-negative diagnoses regarding malignancy, 6 false-positive diagnoses regarding malignancy, and 4 other discordant diagnoses. Eleven of the 29 discordant diagnoses could have had clinical implications. In interpreting these results, it is important to acknowledge the observer variability in diagnostic histopathology in general. In conclusion, the results support the concept of using still images to obtain second opinion diagnosis. PMID- 14745737 TI - Request, respectfully, return to rudiments. PMID- 14745741 TI - Cancer diagnosis and prognosis in Taiwan: patient preferences versus experiences. AB - Considering the emphasis of nonmaleficence and beneficence and the relative power of family, there is a well-recognized suggestion that, in Asian culture, informing cancer patients about their diagnosis and prognosis should be modified according to the family's opinion. However, up until now, except in Japan, the argument that cancer patients from an Asian culture have different preferences about being informed of their diagnosis and prognosis has not been demonstrated by evidence-based studies especially by directly exploring cancer patients' preferences. The purpose of this study was to describe Taiwanese cancer patients' knowledge and experiences of being informed of their diagnosis and prognosis. Preferences of information from health-care professionals and attitudes toward disclosing information to family and even respecting family's opinions not to disclose the life-threatening diagnosis and prognosis to patients have also been explored from the patient's point of views. Evidence from this study demonstrated that Taiwanese cancer patients' information needs are substantial. However, the common practice of nondisclosure of prognosis and detailed disease-related information by health-care professionals continues. Cancer patients in Taiwan expressed a strong preference for health-care professionals to inform them of disease-related information before disclosing information to their family members. Health-care professionals need to consider and respect cancer patients' views rather than routinely taking the family's opinions into consideration first in the event of disagreement. PMID- 14745742 TI - Psychosocial functioning in women who have undergone bilateral prophylactic mastectomy. AB - The purpose of this study was to determine the current psychosocial functioning of women who had previously had a bilateral prophylactic mastectomy. Women in the province of Ontario who had undergone prophylactic mastectomy between 1991 and 2000 were asked to complete questionnaires that assessed psychological distress, sexual activity, overall satisfaction with decision to have a prophylactic mastectomy, and body image. Ninety-seven percent of the women were satisfied with their decision to have a prophylactic mastectomy, but young women (<50 years) were less likely to report satisfaction than older women (p=0.001). Women with a strong family history of breast cancer or a BRCA1 or BRCA2 mutation experienced more cancer-related distress than those with a limited family history. Women who had reconstruction following mastectomy reported higher levels of satisfaction with general body shape and appearance than those without reconstruction. In conclusion, the majority of women were satisfied with their decision to undergo prophylactic mastectomy and were not experiencing abnormal levels of psychological distress, low levels of sexual activity, or difficulties with body image. PMID- 14745743 TI - Illusions in advanced cancer: the effect of belief systems and attitudes on quality of life. AB - Patients with advanced cancer frequently express positive attitudes and can be unduly optimistic about the potential benefits of treatment. In order to evaluate an illusory domain in the context of advanced cancer, we developed a scale of will to live and characterized the beliefs that patients held about the curability of their cancer, and how committed they were to using alternative treatments. A measure of quality of life was used as the dependent variable in order to assess the association between these attributes. After a preliminary exploration confirmed the presence of an illusory domain, these concepts were prospectively tested in 149 ambulant patients with advanced cancer who attended for palliative systemic treatment, radiation treatment or supportive care. The scale of global quality of life was reliable (Cronbach's alpha coefficient 0.72). The distribution of the scores of will to live was skewed, with no respondent scoring poorly, and the scale was reliable (Cronbach's alpha coefficient 0.82). The scale of belief in curability showed diverse beliefs. In some cases, there was a discrepancy between respondents' beliefs in curability and what they believed to be the report by their doctors. There was also an association between a committed use of alternative treatments and a belief in the curability of the cancer (p<0.001). In a multiple regression analysis, both will to live and performance status remained associated with better quality of life scores after adjustment for other relevant variables (p<0.05 and <0.001, respectively). These results suggest that positive illusory beliefs can be measured and are an important component of adaption for some patients with advanced cancer. Furthermore, this illusory domain may influence the perception and measurement of quality of life. PMID- 14745744 TI - Psychoeducational group intervention for wives of men with prostate cancer. AB - OBJECTIVE: The effects of a 6-week psychoeducational group intervention on the distress, coping, personal growth, and marital communication of wives of men diagnosed with prostate cancer were evaluated using a randomized clinical trial. METHODS: Sixty wives completed measures prior to random assignment to either the psychoeducational group intervention or a no-treatment control group, and 1 month after completion of the group. RESULTS: No differences with regard to wives' general distress or cancer-specific distress were noted. In comparison with the control group, participants receiving the intervention perceived that having a spouse with prostate cancer had made positive contributions to their lives, reported gains in the use of positive reappraisal coping and reductions in denial coping. CONCLUSION: Although the psychoeducational intervention did not result in changes in psychological distress, improvements in adaptive coping and indicators of psychological growth were found. The utility of group interventions for spouses of men with prostate cancer is discussed. PMID- 14745745 TI - The effect of dyadic intervention on self-efficacy, social support, and depression for men with prostate cancer. AB - Urinary and sexual dysfunctions are side effects of radical prostatectomy (RP) for prostate cancer (PC) that contribute to depression. Despite the effectiveness of support groups at reducing depression in cancer patients, men typically do not participate in them. The purpose of this pilot study was to test the effects of a dyadic intervention (one-to-one support) on social support (Modified Inventory of Socially Supportive Behaviors), self-efficacy (Stanford Inventory of Cancer Patient Adjustment), and depression (Geriatric Depression Scale). Subjects were randomized to group. Controls (N=15; Mage=59.7) received usual care. Experimentals were paired with long-term survivors (LTS) who had RP and who had treatment side effects in common. Experimentals (N=15; Mage=57.5) met with a LTS 8 times in 8 weeks to discuss concerns associated with survivorship. No significant differences were detected on social support, but after 4 weeks, significant differences were present on depression between controls and experimentals, however these differences were not seen at 8 weeks. After 8 weeks, there were also significant differences on self-efficacy between controls and experimentals. Weekly anecdotal data supported the feasibility and acceptance of the intervention that was a low cost strategy effective at reducing depression and increasing self-efficacy in men treated by RP. Future research directions and clinical application is presented. PMID- 14745746 TI - Does hormone therapy for the treatment of breast cancer have a detrimental effect on memory and cognition? A pilot study. AB - This pilot study examines whether hormone therapy for breast cancer affects cognition. Patients participating in a randomised trial of anastrozole, tamoxifen alone or combined (ATAC) (n=94) and a group of women without breast cancer (n=35) completed a battery of neuropsychological measures. Compared with the control group, the patients were impaired on a processing speed task (p=0.032) and on a measure of immediate verbal memory (p=0.026) after controlling for the use of hormone replacement therapy in both groups. Patient group performance was not significantly related to length of treatment or measures of psychological morbidity. The results showed specific impairments in processing speed and verbal memory in women receiving hormonal therapy for the treatment of breast cancer. Verbal memory may be especially sensitive to changes in oestrogen levels, a finding commonly reported in studies of hormone replacement therapy in healthy women. In view of the increased use of hormone therapies in an adjuvant and preventative setting their impact on cognitive functioning should be investigated more thoroughly. PMID- 14745747 TI - K-space in the clinic. AB - Magnetic resonance imaging (MRI) sequences are characterized by both radio frequency (RF) pulses and time-varying gradient magnetic fields. The RF pulses manipulate the alignment of the resonant nuclei and thereby generate a measurable signal. The gradient fields spatially encode the signals so that those arising from one location in an excited slice of tissue may be distinguished from those arising in another location. These signals are collected and mapped into an array called k-space that represents the spatial frequency content of the imaged object. Spatial frequencies indicate how rapidly an image feature changes over a given distance. It is the action of the gradient fields that determines where in the k-space array each data point is located, with the order in which k-space points are acquired being described by the k-space trajectory. How signals are mapped into k-space determines much of the spatial, temporal, and contrast resolution of the resulting images and scan duration. The objective of this article is to provide an understanding of k-space as is needed to better understand basic research in MRI and to make well-informed decisions about clinical protocols. Four major classes of trajectories-echo planar imaging (EPI), standard (non-EPI) rectilinear, radial, and spiral-are explained. Parallel imaging techniques SMASH (simultaneous acquisition of spatial harmonics) and SENSE (sensitivity encoding) are also described. PMID- 14745748 TI - Blood oxygenation level-dependent MRI of cerebral gliomas during breath holding. AB - PURPOSE: To assess the cerebrovascular responses to short breath holding of cerebral gliomas using blood oxygenation level-dependent (BOLD) magnetic resonance imaging (MRI). MATERIALS AND METHODS: Six patients with a low-grade glioma and one patient with a high-grade glioma were studied using T2*-weighted echo planar imaging (EPI) during repeated periods of 15-second or 20-second breath-holding. Tumor vascularity was evaluated using dynamic susceptibility contrast perfusion MRI. RESULTS: Increases in BOLD signal intensity during repeated breath-holding were consistently identified in patients' normal appearing gray matter, comparable with those in healthy adults. Absence of significant BOLD signal enhancement was noted both in low-grade and high-grade gliomas, which is either due to overwhelming hypoxia in a tumor, inadequacy or absence of hypercapnia-induced vasodilatation of tumor vessels, or both. Breath hold regulated decreases in BOLD signals occurred only in the high-grade glioma, which is most likely due to the hypercapnia-induced steal effect that redistributes blood flow from tumor regions with unresponsive neovasculature to surrounding normal tissue. CONCLUSION: BOLD MRI during short breath holding can disclose differential cerebrovascular response between normal tissue and cerebral glioma. PMID- 14745749 TI - Activation of neural pathways associated with sexual arousal in non-human primates. AB - PURPOSE: To evaluate brain activity associated with sexual arousal, fully conscious male marmoset monkeys were imaged during presentation of odors that naturally elicit high levels of sexual activity and sexual motivation. MATERIAL AND METHODS: Male monkeys were lightly anesthetized, secured in a head and body restrainer with a built-in birdcage resonator and positioned in a 9.4-Tesla spectrometer. When fully conscious, monkeys were presented with the odors of a novel receptive female or an ovariectomized monkey. Both odors were presented during an imaging trial and the presentation of odors was counterbalanced. Significant changes in both positive and negative BOLD signal were mapped and averaged. RESULTS: Periovulatory odors significantly increased positive BOLD signal in several cortical areas: the striatum, hippocampus, septum, periaqueductal gray, and cerebellum, in comparison with odors from ovariectomized monkeys. Conversely, negative BOLD signal was significantly increased in the temporal cortex, cingulate cortex, putamen, hippocampus, substantia nigra, medial preoptic area, and cerebellum with presentation of odors from ovariectomized marmosets as compared to periovulatory odors. A common neural circuit comprising the temporal and cingulate cortices, putamen, hippocampus, medial preoptic area, and cerebellum shared both the positive BOLD response to periovulatory odors and the negative BOLD response to odors of ovariectomized females. CONCLUSION: These data suggest the odor-driven enhancement and suppression of sexual arousal affect neuronal activity in many of the same general brain areas. These areas included not only those associated with sexual activity, but also areas involved in emotional processing and reward. PMID- 14745750 TI - MR flow mapping of dobutamine-induced changes in diastolic heart function. AB - PURPOSE: To evaluate the ability of MR flow mapping to measure changes in left ventricular filling during beta-adrenergic stimulation. MATERIALS AND METHODS: Mitral flow was measured in 10 healthy volunteers using conventional free breathing fast-field echo (FFE) with a spatial resolution of 2.7 x 2.2 mm and a temporal resolution of 22 msec. The sequence was repeated during dobutamine infusion (20 microg/kg/minute). RESULTS: Stroke volume increased from a median of 99 mL (range: 68-142 mL) (Note: values as presented are medians and ranges, throughout) to 114 mL (87-180 mL) (P < 0.05). Both early (E) peak filling rate (554 mL/second [433-497 mL] to 651 mL/second [496-1096 mL/second]) (P < 0.05) and atrial (A) peak filling rate (238 mL/second [183-352 mL/second] to 341 mL/second [230-538 mL/second]) (P < 0.05) increased. These changes, together with the increase in E acceleration peak and A deceleration peak, were consistent with facilitated myocardial relaxation. CONCLUSION: Conventional free breathing FFE has the ability to measure the effects of beta-adrenergic stimulation on left ventricular filling. PMID- 14745751 TI - Transient uterine myometrial contraction associated with moles. AB - PURPOSE: To examine the incidence of transient distortion of uterine central tissue and myometrial hypointense areas observed on MR images in women with clinically suspicious moles. MATERIALS AND METHODS: The study population consisted of six women aged 15-47 years with clinically suspicious moles (hydatidiform mole in four, invasive mole in one, and microscopic mole in one). The control study population was 105 reproductive-age women (18-52 years) without uterine malignancy, gestational trophoblastic disease, or pregnancy. MR images were analyzed to check for discrepancies of the uterine central tissue configuration. If a discrepancy was observed, the myometrial hypointense area, its diameter, and changes in its shape and location were analyzed. RESULTS: Differences in uterine central tissue configuration and hypointense areas were observed in all six patients. In the control study, only seven cases showed uterine endometrial distortion, and five exhibited hypointense areas. These areas disappeared, changed in shape, or other hypointense areas appeared on subsequent MR images. Significant differences (P < 0.01) in the incidence of uterine central tissue distortion and hypointense areas, and in their maximum diameter between the study and the control groups were observed. CONCLUSION: Uterine myometrial hypointense areas with central tissue distortion, most likely due to transient myometrial contraction, are frequently seen in women with clinically suspicious moles. PMID- 14745752 TI - Wall shear rates differ between the normal carotid, femoral, and brachial arteries: an in vivo MRI study. AB - PURPOSE: To investigate wall shear rates in vivo in the common carotid, brachial, and superficial femoral arteries using very high resolution magnetic resonance imaging (MRI) phase contrast measurements. MATERIALS AND METHODS: Mean, maximum, and minimum wall shear rate and an oscillatory shear index were measured for 20 volunteers, aged 23.3 +/- 1.9 years, in the three arteries, using phase contrast imaging with 0.0625 mm2 resolution and three-dimensional paraboloid fitting. RESULTS: The superficial femoral artery had the lowest mean (130.3 +/- 13.1 second(-1)), maximum (735.8 +/- 32.4 second(-1)), and minimum (-224.5 +/- 17.0 second(-1)) wall shear rate, as well as the highest oscillatory shear index (0.21 +/- 0.02). All values were significantly different (P < 0.05) from both the brachial artery and the common carotid artery values. The highest mean (333.3 +/- 13.6 second(-1)) and minimum (117.9 +/- 24.5 second(-1)) wall shear rates and the lowest oscillatory shear index (0 +/- 0) were found in the common carotid artery. CONCLUSION: It is possible to measure wall shear rate in vivo in different arteries using MRI with very high resolution. The findings exhibit the in vivo environment of wall shear rates and suggest a nonuniform distribution of wall shear rates throughout the arterial system. PMID- 14745753 TI - Is half-dose contrast-enhanced three-dimensional MR angiography sufficient for the abdominal aorta and pelvis? AB - PURPOSE: To evaluate the usefulness of half-dose contrast-enhanced magnetic resonance (MR) angiography for depicting the abdominal aorta and its major branches. MATERIALS AND METHODS: A total of 72 consecutive patients were randomly assigned to one of four groups that underwent MR angiography after receiving different concentrations (original or diluted to 50%) and total amounts (single or half-dose) of gadolinium chelate injected at different rates (1 or 0.5 mL/second). The signal-to-noise ratio (SNR) and contrast-to-noise ratio (CNR) of the abdominal aorta and of the common and external iliac arteries were calculated, and two blinded readers rated the respective image qualities. RESULTS: The SNR and CNR of the abdominal aorta and the common iliac artery in the 0.5 mL/second groups were statistically significantly lower than those in the 1 mL/second groups. The differences in overall image quality across the four groups were not statistically significant. CONCLUSION: Half-dose MR angiography using diluted contrast medium injected at a rate of 1 mL/second depicted the abdominal aorta and its branches as clearly as using a full single dose. PMID- 14745754 TI - Time-resolved contrast-enhanced three-dimensional pulmonary MR-angiography: 1.0 M gadobutrol vs. 0.5 M gadopentetate dimeglumine. AB - PURPOSE: To compare contrast characteristics and image quality of 1.0 M gadobutrol with 0.5 M Gd-DTPA for time-resolved three-dimensional pulmonary magnetic resonance angiography (MRA). MATERIALS AND METHODS: Thirty-one patients and five healthy volunteers were examined with a contrast-enhanced time-resolved pulmonary MRA protocol (fast low-angle shot [FLASH] three-dimensional, TR/TE = 2.2/1.0 msec, flip angle: 25 degrees, scan time per three-dimensional data set = 5.6 seconds). Patients were randomized to receive either 0.1 mmol/kg body weight (bw) or 0.2 mmol/kg bw gadobutrol, or 0.2 mmol/kg bw Gd-DTPA. Volunteers were examined three times, twice with 0.2 mmol/kg bw gadobutrol using two different flip angles and once with 0.2 mmol/kg bw Gd-DTPA. All contrast injections were performed at a rate of 5 mL/second. Image analysis included signal-to-noise ratio (SNR) and contrast-to-noise ratio (CNR) measurements in lung arteries and veins, as well as a subjective analysis of image quality. RESULTS: In patients, significantly higher SNR and CNR were observed with Gd-DTPA compared to both doses of gadobutrol (SNR: 35-42 vs.17-25; CNR 33-39 vs. 16-23; P < or = 0.05). No relevant differences were observed between 0.1 mmol/kg bw and 0.2 mmol/kg bw gadobutrol. In volunteers, gadobutrol and Gd-DTPA achieved similar SNR and CNR. A significantly higher SNR and CNR was observed for gadobutrol-enhanced MRA with an increased flip angle of 40 degrees. Image quality was rated equal for both contrast agents. CONCLUSION: No relevant advantages of 1.0 M gadobutrol over 0.5 M Gd-DTPA were observed for time-resolved pulmonary MRA in this study. Potential explanations are T2/T2*-effects caused by the high intravascular concentration when using high injection rates. PMID- 14745755 TI - The utility of MRCP in preoperative mapping of biliary anatomy in adult-to-adult living related liver transplant donors. AB - PURPOSE: To assess the utility of magnetic resonance cholangiopancreatography (MRCP) in preoperative mapping of biliary anatomy in adult-to-adult living related liver transplant (LRLT) donors. MATERIALS AND METHODS: From 57 potential donors with preoperative MRCP, 27 cases (16 men, 11 women, age range 22-51 years, mean 37.2 years) underwent right lobe resection and had intraoperative cholangiography (IOC) for comparison. The MRCP and IOC reports were retrospectively reviewed in all 27 cases. The MRCP was performed on 1.5 Tesla MR magnets using breath-hold heavily T2-weighted sequences in axial/coronal thin sections, and variable-thickness rotating slabs. The accuracy of preoperative MRCP for biliary mapping in potential LRLT donors was analyzed compared to the IOC findings. RESULTS: Of 27 donors, 26 (96.3%) had MRCP which showed adequate information of central intrahepatic biliary anatomy. Of these, 19 had normal bifurcation confirmed by IOC, and single biliary anastomosis was created in the recipient at transplantation. MRCP correctly predicted 17 of 19 normal cases (sensitivity for normals: 89.5%). In seven donors with variant biliary anatomy, two separate biliary anastomoses were performed in the recipient. MRCP correctly predicted five of seven variants (sensitivity for variants: 71.4%). Overall, MRCP had an accuracy 84.6% (22/26). CONCLUSIONS: MRCP has potential in the preoperative assessment of nondilated bile ducts in LRLT donors, however further improvements are desired to increase its quality and accuracy. PMID- 14745756 TI - MR colonography: how does air compare to water for colonic distention? AB - PURPOSE: To prove the feasibility of air-distended magnetic resonance colonography (MRC) and compare it with water-based distention. MATERIALS AND METHODS: In five volunteers, the colon was imaged twice: once after distending the colon with air and a second time after distending the colon with water. A total of 50 patients, who had been referred to colonoscopy for a suspected colorectal pathology were randomized into water-distention (N = 25) and air distention (N = 25) groups. A contrast-enhanced T1-weighted three-dimensional volume interpolated breath-hold (VIBE) sequence was collected. Comparative analysis was based on qualitative ratings of image quality and bowel distention, as well as contrast-to-noise ratio (CNR) measurements for the colonic wall with respect to the colonic lumen. In addition, patient acceptance was evaluated. RESULTS: Inflammatory changes and colorectal masses were correctly identified on MRC in eight patients each. One 4-mm polyp identified at colonoscopy was missed on water-distended MRC. There were no false positive findings. No significant differences were found between air- and water-distention regarding discomfort levels and image quality. The presence of air in the colonic lumen was not associated with susceptibility artifacts. CNR of the contrast-enhanced colonic wall, as well as bowel distention, were superior on air-distended three dimensional data sets. CONCLUSION: MRC can be performed using either water or air for colonic distention. Both techniques permit assessment of the colonic wall and identification of colorectal masses. While discomfort levels are similar for both agents, MRC with air provides higher CNR and better colonic distention. PMID- 14745757 TI - Bone marrow diffusion in osteoporosis: evaluation with quantitative MR diffusion imaging. AB - PURPOSE: To determine the diffusion of vertebral body marrow with quantitative MR diffusion imaging and to examine whether differences exist between subjects with postmenopausal osteoporosis and premenopausal control subjects. MATERIALS AND METHODS: A total of 44 consecutive women (mean age, 70 years) with documented bone mineral density (BMD) measured by dual energy x-ray absorptiometry (T-score) and 20 normal subjects (mean age, 28 years) were examined with echo-planar diffusion imaging at 1.5 T using b values of 0, 20, 40, 60, 80, 100, 200, 300, 400, and 500 seconds/mm2. Extravascular diffusion (D) and apparent diffusion coefficient (ADC) were calculated and results from both groups compared. RESULTS: Both D and ADC values tended to decrease with decreasing BMD. Mean D values were significantly lower in postmenopausal women with reduced BMD (0.42 +/- 0.12 x 10( 3) mm2/second) than normal premenopausal women (0.50 +/- 0.09 x 10(-3) mm2/second). Mean ADC values were significantly lower both in subjects with reduced BMD (0.41 +/- 0.10 x 10(-3) mm2/second) and normal BMD (0.43 +/- 0.08 x 10(-3) mm2/second) compared to normal controls (0.49 +/- 0.07 x 10(-3) mm2/second). CONCLUSION: Accumulation of fatty bone marrow associated with osteoporosis is reflected by a decrease in D and ADC. Diffusion imaging may prove useful in the study of osteoporosis. PMID- 14745758 TI - Assessment of motion gating strategies for mouse magnetic resonance at high magnetic fields. AB - PURPOSE: To assess the performance of motion gating strategies for mouse cardiac magnetic resonance (MR) at high magnetic fields by quantifying the levels of motion artifact observed in images and spectra in vivo. MATERIALS AND METHODS: MR imaging (MRI) of the heart, diaphragm, and liver; MR angiography of the aortic arch; and slice-selective 1H-spectroscopy of the heart were performed on anesthetized C57Bl/6 mice at 11.75 T. Gating signals were derived using a custom built physiological motion gating device, and the gating strategies considered were no gating, cardiac gating, conventional gating (i.e., blanking during respiration), automatic gating, and user-defined gating. Both automatic and user defined modes used cardiac and respiratory gating with steady-state maintenance during respiration. Gating performance was assessed by quantifying the levels of motion artifact observed in images and the degree of amplitude and phase stability in spectra. RESULTS: User-defined gating with steady-state maintenance during respiration gave the best performance for mouse cardiac imaging, angiography, and spectroscopy, with a threefold increase in signal intensity and a sixfold reduction in noise intensity compared to cardiac gating only. CONCLUSION: Physiological gating with steady-state maintenance during respiration is essential for mouse cardiac MR at high magnetic fields. PMID- 14745759 TI - Magnetic resonance portography using contrast-enhanced fat-saturated three dimensional steady-state free precession imaging. AB - PURPOSE: To assess the feasibility of contrast-enhanced fat-saturated three dimensional steady-state free precession (FIESTA) imaging for contrast-enhanced magnetic resonance (MR) portography. MATERIALS AND METHODS: Contrast-enhanced fat saturated three-dimensional fast spoiled gradient-echo (SPGR) and FIESTA were performed as MR portography. In 10 cases, fat-saturated three-dimensional FIESTA was first performed and followed by fast SPGR, and the order of post-contrast imaging was reversed in the other 10 cases. Signal-to-noise ratios (SNRs) and contrast-to-noise ratios (CNRs) were estimated for portal and visceral veins on the source images. The visualization of portal vein was scored on three dimensional MR portography. Portal venous system disorders were assessed using three-dimensional MR portography. RESULTS: The SNRs, CNRs, and visual assessment of portal and visceral veins were significantly higher in contrast-enhanced fat saturated three-dimensional FIESTA than contrast-enhanced fat-saturated three dimensional fast SPGR (P < 0.05). The contrast-enhanced fat-saturated three dimensional FIESTA provided high venous signals even at 8 minutes after gadolinium injection. The abnormalities of portal venous system were well visualized with MR portography using contrast-enhanced fat-saturated three dimensional FIESTA. CONCLUSION: Contrast-enhanced fat-saturated three-dimensional FIESTA was valuable for MR portography, with flexible time window and high vascular signals. This imaging may allow for other post-contrast imaging options before portography and release patients from consecutive breath-holds. PMID- 14745760 TI - Automatic model-based evaluation of magnetic resonance-guided radio frequency ablation lesions with histological correlation. AB - PURPOSE: To develop a model-based method for automatic evaluation of radio frequency (RF) ablation treatment using magnetic resonance (MR) images. MATERIALS AND METHODS: RF current lesions were generated in a rabbit thigh model using MR imaging (MRI) guidance. We created a 12-parameter, three-dimensional, globally deformable model with quadric surfaces that delineates lesion boundaries and is automatically fitted to MR grayscale data. We applied this method to in vivo T2- and contrast-enhanced (CE) T1-weighted MR images acquired immediately post ablation and four days later. We then compared results to manually segmented MR and three-dimensional registered corresponding histological boundaries of cellular damage. RESULTS: Resulting lesions featured a two-boundary appearance with an inner region and an outer hyperintense margin on MR images. For automated vs. manual MR boundaries, the mean errors over all specimens were 0.19 +/- 0.51 mm and 0.27 +/- 0.52 mm for the inner surface, and -0.29 +/- 0.40 mm and -0.12 +/ 0.17 mm for the outer surface, for T2- and CE T1-weighted images, respectively. For automated vs. histological boundaries, mean errors over all specimens were 0.07 +/- 0.64 mm and 0.33 +/- 0.71 mm for the inner surface, and -0.27 +/- 0.69 mm and 0.02 +/- 0.43 mm for the outer surface, for T2- and CE T1-weighted images, respectively. All boundary errors compared favorably to MR voxel dimensions, which were 0.7 mm in-plane and 3.0 mm thick. CONCLUSION: The method is accurate both in describing MR-apparent boundaries and in predicting histological response and has applications in lesion visualization, volume estimation, and treatment evaluation. PMID- 14745761 TI - A method for preamplifier-decoupling improvement in quadrature phased-array coils. AB - PURPOSE: To improve preamplifier decoupling in a quadrature phased-array coil using a new quadrature combiner configuration. MATERIALS AND METHODS: A commercial six-element quadrature cervical-thoracic-lumbar (CTL) coil was used on a 1.5 T GE Signa Lx Scanner. The new combiner configuration is designed based on the fact that when one compares the amount of flux linkage of two loops with butterfly elements separated by the same distance, the loop elements have much higher coupling than the butterfly elements. RESULTS: The results are demonstrated using an imaging-based element coupling measurement method. The new quadrature combiner configuration has introduced an approximately -8dB improvement in preamplifier decoupling. CONCLUSION: One-dimensional quadrature phased arrays can be built even if the preamplifier is not integrated in the coil. If the combiner is placed between the coil and the preamplifiers, the butterflies should be on the high impedance side of the quadrature combiner. The loops should see the low input impedance of the preamplifiers. PMID- 14745762 TI - Qualitative and quantitative analysis of low molecular weight compounds by ultraviolet matrix-assisted laser desorption/ionization mass spectrometry using ionic liquid matrices. AB - A major problem hampering the use of matrix-assisted laser desorption/ionization (MALDI) mass spectrometry for quantitative measurements is the inhomogeneous distribution of analytes and matrices in solid sample preparations. The use of ionic liquids as matrices for the qualitative and quantitative analysis of low molecular weight compounds like amino acids, sugars and vitamins was investigated. The ionic liquid matrices are composed of equimolar combinations of classical MALDI matrices (sinapinic acid, alpha-cyano-4-hydroxycinnamic acid or 2,5-dihydroxybenzoic acid) with organic bases. These matrix systems allow a homogenous sample preparation with a thin ionic liquid layer having negligible vapour pressure. This leads to a facilitated qualitative and quantitative measurement of the analytes compared with classical solid matrices. PMID- 14745764 TI - Solid-phase microextraction combined with surface-enhanced laser desorption/ionization introduction for ion mobility spectrometry and mass spectrometry using polypyrrole coatings. AB - The successful application of polypyrrole (PPY) solid-phase microextraction (SPME) coatings as both an extraction phase and a surface to enhance laser desorption/ionization (SELDI) of analytes is reported. This SPME/SELDI fiber integrates sample preparation and sample introduction on the tip of a coated optical fiber, as well as acting as the transmission medium for the UV laser light. Using ion mobility spectrometry (IMS) detection, the signal intensity was examined as a function of extraction surface area and concentration of analyte. The linear relationship between concentration and signal intensity shows potential applicability of this detection method for quantitative analysis. Extraction time profiles for the fiber, using tetraoctylammonium bromide as test analyte, illustrated that equilibrium can be reached in less than one minute. To investigate the performance of the PPY coating, the laser desorption profile was studied. The fiber was also tested using a quadrupole time-of-flight (Q-TOF) mass spectrometer with leucine enkephalin as test analyte. Since no matrix was used, mass spectra free from matrix background were obtained. This novel SPME/SELDI fiber is easy to manufacture, and is suitable for studying low-mass analytes because of the intrinsic low background. These findings suggest that other types of conductive polymers could also be used as an extraction phase and surface to enhance laser desorption/ionization in mass spectrometry. PMID- 14745763 TI - Mass spectrometry for the detection of differentially expressed proteins: a comparison of surface-enhanced laser desorption/ionization and capillary electrophoresis/mass spectrometry. AB - The discovery of biomarkers is currently attracting much interest as it harbors great potential for the diagnosis and monitoring of human diseases. Here we have used two advanced mass spectroscopy based technologies, surface enhanced laser desorption ionization (SELDI-MS) and capillary electrophoresis/mass spectrometry (CE/MS), to obtain proteomic patterns of urine samples from patients suffering from membranous glomerulonephritis (MGN) and healthy volunteers. The results indicate that CE/MS analysis is able to display a rich and complex pattern of polypeptides with high resolution and high mass accuracy. In order to analyze these patterns, the MosaiqueVisu software was developed for peak identification, deconvolution and the display of refined maps in a three-dimensional format. The polypeptide profiles obtained with SELDI-MS from the same samples are much sparser and show lower resolution and mass accuracy. The SELDI-MS profiles are further heavily dependent on analyte concentration. SELDI-MS analysis identified three differentially expressed polypeptides, which are potential biomarkers that can distinguish healthy donors from patients with MGN. In contrast, approximately 200 potential biomarkers could be identified by CE/MS. Thus, while SELDI-MS is easy to use and requires very little sample, CE/MS generates much richer data sets that enable an in-depth analysis. PMID- 14745765 TI - Improvements and new capabilities for the multiple Knudsen cell device used in high-temperature mass spectrometry. AB - The thermodynamic properties of condensed phases, i.e. the activities of components, can be determined from partial pressures measured by the Knudsen cell mass spectrometric method. Improvements in accuracy and yield of this method are obtained with the use of twin cells, an idea proposed in the 1960s. The multiple cell method was perfected in 1977 in our laboratory. Changes to molecular beam sampling and furnace assemblies were required to make the multiple Knudsen cell technique work properly. This paper summarizes these prerequisites, and presents a new device and the associated method of measurements, as well as the necessary tests performed with a silver sample in each cell. PMID- 14745766 TI - A new method for determining delta13C and deltaD simultaneously for CH4 by gas chromatography/continuous-flow isotope-ratio mass spectrometry. AB - A continuous-flow technique has been developed to analyse the deltaD and delta(13)C values for CH(4) from gas samples, in a single run. This is achieved by splitting the sample gas stream and directing the streams simultaneously through a CuNiPt combustion reactor and an alumina pyrolysis reactor. The CO(2) from CH(4) combustion is trapped in a liquid nitrogen trap while the H(2) exiting the pyrolysis reactor is directed to the mass spectrometer for deltaD(CH4) determination. The CO(2) is then sublimed and directed to the mass spectrometer for delta(13)C(CH4) determination. Sample runs take approximately 10 minutes. This technique gives accurate delta(13)C(CH4) results to within +/-0.3-0.5 per thousand and deltaD(CH4) results to within +/-2-5 per thousand. Injection volumes between 0.5 and 2.5 microL of CH(4), equivalent to between 20 and 100 nmol CH(4), are required for accurate delta(13)C and deltaD analyses, respectively, using sample injection into a split flow with a split ratio of 10. This method provides rapid, accurate and reproducible results on multiple sample runs and is, therefore, an ideal method for analysing natural gas samples from a variety of sources. PMID- 14745767 TI - Sensitive sulphur-specific detection of omeprazole metabolites in rat urine by high-performance liquid chromatography/inductively coupled plasma mass spectrometry. AB - The use of high-performance liquid chromatography/inductively coupled plasma mass spectrometry (HPLC/ICPMS) with sulphur-specific detection was investigated as a method for obtaining metabolite profiles for the drug omeprazole administered as a 1:1 mixture of (32)S- and (34)S-labelled material. Analysis based on the monitoring of the chromatographic eluent at either m/z 32 or 34 was not successful due to insufficient sensitivity caused by interferences from polyatomic ions. However, reaction of sulphur with oxygen in the hexapole collision cell, combined with monitoring at m/z 48 (for (32)S) or m/z 50 (for (34)S), provided a facile method for metabolite profiling. Detection of m/z 48 was superior in sensitivity to detection of m/z 50. PMID- 14745768 TI - On-line identification of phenanthroindolizidine alkaloids in a crude extract from Tylophora atrofolliculata by liquid chromatography combined with tandem mass spectrometry. AB - Electrospray ionization multi-stage tandem mass spectrometry (ESI-MS(n)) and liquid chromatography coupled with sequential mass spectrometry (LC/MS(n)) were applied to identify trace-level phenanthroindolizidine alkaloids in crude extracts from Tylophora atrofolliculata. Based on the relationship between the characteristic fragmentation reactions and the structural features of related compounds of known structure from this plant, the bioactive crude extract was analyzed in detail by positive and negative ion ESI-MS(n), LC/UV-MS and LC/MS(n) techniques. A total of nine constituents in the crude extract were identified rapidly, including several isomers; seven of these constituents are new and two are known compounds. The structures of four of these constituents were subsequently confirmed by nuclear magnetic resonance (NMR) and accurate mass measurements using high-resolution fast-atom bombardment mass spectrometry (FAB HRMS). PMID- 14745769 TI - De novo sequencing of tryptic peptides sulfonated by 4-sulfophenyl isothiocyanate for unambiguous protein identification using post-source decay matrix-assisted laser desorption/ionization mass spectrometry. AB - A simple method of solid-phase derivatization and sequencing of tryptic peptides has been developed for rapid and unambiguous identification of spots on two dimensional gels using post-source decay (PSD) matrix-assisted laser desorption/ionization (MALDI) mass spectrometry. The proteolytic digests of proteins are chemically modified by 4-sulfophenyl isothiocyanate. The derivatization reaction introduces a negative sulfonic acid group at the N terminus of a peptide, which can increase the efficiency of PSD fragmentation and enable the selective detection of only a single series of fragment ions (y-ions). This chemically assisted method avoids the limitation of high background normally observed in MALDI-PSD spectra, and makes the spectra easier to interpret and facilitates de novo sequencing of internal fragment. The modification reaction is conducted in C(18) microZipTips to decrease the background and to enhance the signal/noise. Derivatization procedures were optimized for MALDI-PSD to increase the structural information and to obtain a complete peptide sequence even in critical cases. The MALDI-PSD mass spectra of two model peptides and their sulfonated derivatives are compared. For some proteins unambiguous identification could be achieved by MALDI-PSD sequencing of derivatized peptides obtained from in-gel digests of phosphorylase B and proteins of hepatic satellite cells (HSC). PMID- 14745770 TI - Determination of specific volatile organic compounds synthesised during Tuber borchii fruit body development by solid-phase microextraction and gas chromatography/mass spectrometry. AB - Fruit body development is a particular phase of the Tuber life cycle, characterised by the aggregation of different types of hyphae, i.e., vegetative hyphal cells and highly specialised reproductive hyphae (asci). In order to identify the volatile organic compounds (VOCs) produced in different stages of the Tuber borchii ripening fruit body, solid-phase microextraction with gas chromatography and mass spectrometry was used. The volatile organic compounds were extracted using a DVB/CAR/PDMS 50/30 microm fiber placed for 10 min at room temperature in the truffle headspace. The results obtained reveal 49 compounds each of which was present only in a particular stage of maturation. 1-octen-3-ol, aromadendrene, alpha-farnesene and other terpenoid compounds were of particular interest, and their possible biological roles are discussed. The production of aromadendrene in the completely unripe fruit body suggests the existence of communication events in the early stage of ascomata formation between the fungus and the host plant. alpha-Farnesene could represent a chemotactic attractant to saprophytic organisms in order to disperse the fungal spores in the environment. The identification of the VOCs produced by truffles during their maturation could give information about the processes underlying this phase of Tuber life cycle. PMID- 14745771 TI - Headspace solid-phase microextraction with gas chromatography and mass spectrometry in the investigation of volatile organic compounds in an ectomycorrhizae synthesis system. AB - Ectomycorrhizae formation represents one of the most significant steps in the truffle life cycle and is determined by a complex molecular signaling between two symbionts. In order to understand the molecular pathway of ectomycorrhiza development, we focused on the signaling interaction between the ectomycorrhizal fungus Tuber borchii Vittad. and the Tilia americana L. plant roots. The medium of a pre-symbiotic (T. americana-T. borchii) in vitro system was analysed by headspace solid-phase microextraction coupled with gas chromatography and mass spectrometry. In total, 73 volatile organic compounds (VOCs) were identified. Twenty-nine of these VOCs were produced only during the interaction phase between the two partners, leading to a hypothesis that these molecules might act as molecular messengers in order to pilot the ectomycorrhizae formation. PMID- 14745772 TI - Simultaneous co-extraction of organometallic species of different elements by accelerated solvent extraction and analysis by inductively coupled plasma mass spectrometry coupled to liquid and gas chromatography. AB - This paper describes the development of an accelerated solvent extraction methodology that is capable of simultaneously extracting organometallic species of As, Sn and Hg in a semi-automated manner. Accelerated solvent extraction (ASE) methods based on previous research on the separate extraction of organotin and arsenic species in our laboratory were adapted for the co-extraction of six different species from an oyster tissue certified reference material (BCR 710). For the first time, the extraction of MeHg by this technique is also investigated. The proposed ASE conditions employed 50% acetic acid in methanol at a temperature of 100 degrees C with up to five consecutive extraction cycles of 3 minutes. Extraction efficiencies for organoarsenic species ranged from 80% (dimethylarsinic acid, DMA) to 99% for arsenobetaine (AsB). Species of toxicological interest, such as dibutyltin (DBT), tributyltin (TBT) and methylmercury (MeHg), were extracted with mean recoveries of 81, 84 and 76%, respectively. The extracted species were analysed by gas chromatography/inductively coupled plasma mass spectrometry (GC/ICPMS; DBT, TBT and MeHg) and liquid chromatography/inductively coupled plasma mass spectrometry (LC/ICPMS; MMA, DMA, AsB) after ethylation with sodium tetraethylborate or dilution with water, respectively. In addition to those species for which the extraction efficiency was assessed during this study, a further five arsenic species (arsenite, arsenate and three unidentified species), as well as monobutyltin (MBT) and mono-, di- and triphenyltin, could also be extracted from other matrices. The developed ASE method provides a significant improvement over many currently available routine monitoring methods for trace element speciation due to the fact that it is capable of extracting several species of toxicological interest simultaneously and quantitatively. PMID- 14745773 TI - Quantitative analysis of triacylglycerol regioisomers in fats and oils using reversed-phase high-performance liquid chromatography and atmospheric pressure chemical ionization mass spectrometry. AB - Positional distribution of fatty acyl chains of triacylglycerols (TGs) in vegetable oils and fats (palm oil, cocoa butter) and animal fats (beef, pork and chicken fats) was examined by reversed-phase high-performance liquid chromatography (RP-HPLC) coupled to atmospheric pressure chemical ionization using a quadrupole mass spectrometer. Quantification of regioisomers was achieved for TGs containing two different fatty acyl chains (palmitic (P), stearic (S), oleic (O), and/or linoleic (L)). For seven pairs of 'AAB/ABA'-type TGs, namely PPS/PSP, PPO/POP, SSO/SOS, POO/OPO, SOO/OSO, PPL/PLP and LLS/LSL, calibration curves were established on the basis of the difference in relative abundances of the fragment ions produced by preferred losses of the fatty acid from the 1/3 position compared to the 2-position. In practice the positional isomers AAB and ABA yield mass spectra showing a significant difference in relative abundance ratios of the ions AA(+) to AB(+). Statistical analysis of the validation data obtained from analysis of TG standards and spiked oils showed that, under repeatability conditions, least-squares regression can be used to establish calibration curves for all pairs. The regression models show linear behavior that allow the determination of the proportion of each regioisomer in an AAB/ABA pair, within a working range from 10 to 1000 microg/mL and a 95% confidence interval of +/-3% for three replicates. PMID- 14745774 TI - Detection of technical chlordane by liquid chromatography with atmospheric pressure chemical ionization mass spectrometry. PMID- 14745775 TI - A nitrocellulose matrix for infrared matrix-assisted laser desorption/ionization of polycyclic aromatic hydrocarbons. PMID- 14745776 TI - Determination of boron isotope ratio in boron carbide using a laser mass spectrometric method. PMID- 14745777 TI - Multi-stage mass spectrometry of furostanol saponins combined with electrospray ionization in positive and negative ion modes. PMID- 14745778 TI - Isolation and characterization of Candida albicans homologue of RAP1, a repressor and activator protein gene in Saccharomyces cerevisiae. AB - To study the function of RAP1, a Candida albicans gene (CaRAP1) that shows sequence similarity to RAP1 of Saccharomyces cerevisiae was isolated by colony hybridization. DNA sequencing predicted an open reading frame of 429 amino acids with an overall identity of 24% to the ScRap1p. The DNA binding domain (DBD) was highly conserved, and EMSA using a GST-CaRap1p fusion protein confirmed its binding ability to the RPG-box of S. cerevisiae ENO1. In contrast, the N-terminus was less conserved and a moderate homology was observed in the BRCT domain. Interestingly, CaRap1p did not contain the C-terminal activation/repression region of ScRap1p. PMID- 14745779 TI - Identification of an UDP-Glc:glycoprotein glucosyltransferase in the yeast Yarrowia lipolytica. AB - The UDP-Glc:glycoprotein glucosyltransferase (UGT) is a soluble protein of the endoplasmic reticulum (ER) that plays a determining part in the mechanism by which unfolded, partially folded or misfolded glycoproteins are retained into the ER. We have identified an UGT in the yeast Yarrowia lipolytica. This protein, of a predicted molecular weight of 165.7 kDa, is encoded by a 5054 bp coding sequence containing a 643 bp intron at position 682-1323. The N-terminal part of the protein displays a signal sequence whereas its C-terminal part carries an ER retrieval signal HDEL. An interruption of the gene that removes the 1075 last nucleotides of its sequence did not lead to any evident phenotype except for a slight increased sensitivity to tunicamycin. YlUGT1 mRNA levels respond to tunicamycin treatment by an induction factor of 2-4, which indicates that the gene product participates in the quality control mechanism in this yeast. Finally, an immunofluorescence study of the protein localization, shows that the protein distribution is different from that of previously studied ER resident proteins. This could indicate that UGT distribution in the secretory pathway is not confined to the ER. PMID- 14745780 TI - A DNA microarray for fission yeast: minimal changes in global gene expression after temperature shift. AB - Completion of the fission yeast genome sequence has opened up possibilities for post-genomic approaches. We have constructed a DNA microarray for genome-wide gene expression analysis in fission yeast. The microarray contains DNA fragments, PCR-amplified from a genomic DNA template, that represent > 99% of the 5000 or so annotated fission yeast genes, as well as a number of control sequences. The GenomePRIDE software used attempts to design similarly sized DNA fragments corresponding to gene regions within single exons, near the 3'-end of genes that lack homology to other fission yeast genes. To validate the design and utility of the array, we studied expression changes after a 2 h temperature shift from 25 degrees C to 36 degrees C, conditions widely used when studying temperature sensitive mutants. Obligingly, the vast majority of genes do not change more than two-fold, supporting the widely held view that temperature-shift experiments specifically reveal phenotypes associated with temperature-sensitive mutants. However, we did identify a small group of genes that showed a reproducible change in expression. Importantly, most of these corresponded to previously characterized heat-shock genes, whose expression has been reported to change after more extreme temperature shifts than those used here. We conclude that the DNA microarray represents a useful resource for fission yeast researchers as well as the broader yeast community, since it will facilitate comparison with the distantly related budding yeast, Saccharomyces cerevisiae. To maximize the utility of this resource, the array and its component parts are fully described in On-line Supplementary Information and are also available commercially. PMID- 14745781 TI - KlSEC53 is an essential Kluyveromyces lactis gene and is homologous with the SEC53 gene of Saccharomyces cerevisiae. AB - Phosphomannomutase (PMM) is a key enzyme, which catalyses one of the first steps in the glycosylation pathway, the conversion of D-mannose-6-phosphate to D mannose-1-phosphate. The latter is the substrate for the synthesis of GDP mannose, which serves as the mannosyl donor for the glycosylation reactions in eukaryotic cells. In the yeast Saccharomyces cerevisiae PMM is encoded by the gene SEC53 (ScSEC53) and the deficiency of PMM activity leads to severe defects in both protein glycosylation and secretion. We report here on the isolation of the Kluyveromyces lactis SEC53 (KlSEC53) gene from a genomic library by virtue of its ability to complement a Saccharomyces cerevisiae sec53 mutation. The sequenced DNA fragment contained an open reading frame of 765 bp, coding for a predicted polypeptide, KlSec53p, of 254 amino acids. The KlSec53p displays a high degree of homology with phosphomannomutases from other yeast species, protozoans, plants and humans. Our results have demonstrated that KlSEC53 is the functional homologue of the ScSEC53 gene. Like ScSEC53, the KlSEC53 gene is essential for K. lactis cell viability. Phenotypic analysis of a K. lactis strain overexpressing the KlSEC53 gene revealed defects expected for impaired cell wall integrity. PMID- 14745782 TI - Positive and negative selection LYS5MX gene replacement cassettes for use in Saccharomyces cerevisiae. AB - Novel MX cassettes are described that contain the open reading frames (ORFs) of Saccharomyces cerevisiae or Candida albicans LYS5. The LYS5MX and CaLYS5MX cassettes, the targeting efficiencies of which are equivalent to those of other MX cassettes, are positively selected for Lys+ in a lys5 background. Unlike most of the other MX cassettes, the LYS5MX cassettes are also negatively selectable (alpha-aminoadipate-resistant), which will allow the use of the LYS5MX cassettes in plasmid shuffling and will also greatly facilitate marker recycling. PMID- 14745783 TI - Alanine : glyoxylate aminotransferase of Saccharomyces cerevisiae-encoding gene AGX1 and metabolic significance. AB - Alanine : glyoxylate aminotransferase is one of three different enzymes used for glycine synthesis in Saccharomyces cerevisiae. The open reading frame YFL030w (named AGX1 in the following), encoding this enzyme, was identified by comparing enzyme specific activities in knockout strains. While 100% activity was detectable in the parental strain, 2% was found in a YFL030w::kanMX4 strain. The ORF found at that locus was suspected to encode alanine : glyoxylate aminotransferase because its predicted amino acid sequence showed 23% identity to the human homologue. Since the YFL030w::kanMX4 strain showed no glycine auxtrophic phenotype, AGX1 was replaced by KanMX4 in a Delta GLY1 Delta SHM1 Delta SHM2 background. These background mutations, which cause inactivation of threonine aldolase, mitochondrial and cytosolic serine hydroxymethyltransferase, respectively, lead to a conditional glycine auxotrophy. This means that growth is not possible on glucose but on ethanol as the sole carbon source. Additional disruption of AGX1 revealed a complete glycine auxotrophy. Complementation was observed by transformation with a plasmid-encoded AGX1. PMID- 14745784 TI - The high general stress resistance of the Saccharomyces cerevisiae fil1 adenylate cyclase mutant (Cyr1Lys1682) is only partially dependent on trehalose, Hsp104 and overexpression of Msn2/4-regulated genes. AB - The initiation of fermentation in the yeast Saccharomyces cerevisiae is associated with a rapid drop in general stress resistance. Previously we identified a mutant which is deficient in fermentation-induced loss of stress resistance (fil1), as a partially inactivating mutant in adenylate cyclase. We have now investigated possible causes of its high stress resistance. Deletion of the TPS1 gene, encoding the first enzyme in the biosynthesis of trehalose, or the heat shock protein gene HSP104 only resulted in a minor effect on heat stress resistance compared with deletion of these genes in a wild-type background. A strain with a deletion of both genes still showed a higher stress resistance in the fil1 background compared to the corresponding wild-type background. Deletion of the transcription factor genes MSN2 and MSN4, which are required for the expression of STRE-regulated genes, resulted in a dramatic drop in heat resistance in the wild-type background but had much less effect in the fil1 mutant. The fil1 msn2Deltamsn4Delta strain remained more heat-resistant than a wild-type strain. A strain in which all four genes, TPS1, HSP104, MSN2 and MSN4, are deleted was very sensitive to heat stress and also to oxidative and salt stress. Presence of the fil1 mutation in such a strain, however, still clearly enhanced heat, oxidative and salt stress resistance. These results indicate that, in addition to trehalose, Hsp104 and the Msn2/4-controlled genes, other factors exist in S. cerevisiae that can, significantly and independently of the known factors, enhance general stress resistance. The mutants described in this work provide a tool to identify these novel components. PMID- 14745785 TI - Editorial. PMID- 14745788 TI - Conformational response of the phosphatidylcholine headgroup to bilayer surface charge: torsion angle constraints from dipolar and quadrupolar couplings in bicelles. AB - The effects of bilayer surface charge on the conformation of the phosphocholine group of phosphatidylcholine were investigated using a torsion angle analysis of quadrupolar and dipolar splittings in, respectively, (2)H and (13)C NMR spectra of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) labelled in the phosphocholine group with either deuterons (POPC-alpha-d(2), POPC-beta-d(2) and POPC-gamma-d(9)) or carbon-13 (POPC-alpha-(13)C and POPC-alphabeta-(13)C(2)) and incorporated into magnetically aligned bicelles containing various amounts of either the cationic amphiphile 1,2-dimyristoyl-3-trimethylammoniumpropane (DMTAP) or the anionic amphiphile 1,2-dimyristoyl-sn-glycero-3-phosphoglycerol (DMPG). Three sets of quadrupolar splittings, one from each of the three deuteron labelling positions, and three sets of dipolar splittings ((13)C(alpha)-(31)P, (13)C(alpha)-(13)C(beta), (13)C(beta)-(14)N), were measured at each surface charge, along with the (31)P residual chemical shift anisotropy. The torsion angle analysis assumed fast anisotropic rotation of POPC about its long molecular axis, thus projecting all NMR interactions onto that director axis of motion. Dipolar, quadrupolar and chemical shift anisotropies were calculated as a function of the phosphocholine internal torsion angles by first transforming into a common reference frame affixed to the phosphocholine group prior to motional averaging about the director axis. A comparison of experiment and calculation provided the two order parameters specifying the director orientation relative to the molecule, plus the torsion angles alpha(3), alpha(4) and alpha(5). Surface charge was found to have little effect on the torsion angle alpha(5) (rotations about C(alpha)-C(beta)), but to have large and inverse effects on torsion angles alpha(3) [rotations about P-O(11)] and alpha(4) [rotations about O(11)-C(alpha)], yielding a net upwards tilt of the P-N vector in the presence of cationic surface charge, and a downwards tilt in the presence of anionic surface charge, relative to neutrality. PMID- 14745789 TI - Effects of antidepressants on the conformation of phospholipid headgroups studied by solid-state NMR. AB - The effect of tricyclic antidepressants (TCA) on phospholipid bilayer structure and dynamics was studied to provide insight into the mechanism of TCA-induced intracellular accumulation of lipids (known as lipidosis). Specifically we asked if the lipid-TCA interaction was TCA or lipid specific and if such physical interactions could contribute to lipidosis. These interactions were probed in multilamellar vesicles and mechanically oriented bilayers of mixed phosphatidylcholine-phosphatidylglycerol (PC-PG) phospholipids using (31)P and (14)N solid-state NMR techniques. Changes in bilayer architecture in the presence of TCAs were observed to be dependent on the TCA's effective charge and steric constraints. The results further show that desipramine and imipramine evoke distinguishable changes on the membrane surface, particularly on the headgroup order, conformation and dynamics of phospholipids. Desipramine increases the disorder of the choline site at the phosphatidylcholine headgroup while leaving the conformation and dynamics of the phosphate region largely unchanged. Incorporation of imipramine changes both lipid headgroup conformation and dynamics. Our results suggest that a correlation between TCA-induced changes in bilayer architecture and the ability of these compounds to induce lipidosis is, however, not straightforward as imipramine was shown to induce more dramatic changes in bilayer conformation and dynamics than desipramine. The use of (14)N as a probe was instrumental in arriving at the presented conclusions. PMID- 14745790 TI - Multi-dimensional pulsed field gradient magic angle spinning NMR experiments on membranes. AB - The benefits of gradient techniques in the study of lipid membranes are demonstrated on a sample of 1-palmitoyl-2-oleoyl-sn-glycero-3 phosphocholine (POPC) liposomes embedded with ibuprofen. Most techniques from gradient NMR spectroscopy on solution samples are directly applicable to membrane samples subjected to magic angle spinning (MAS). Gradient-enhanced homo- and heteronuclear chemical shift correlation techniques were used to make resonance assignments. Gradient NOESY experiments provide insight into the location and dynamics of lipids, ibuprofen and water. Application of gradients not only reduces experiment time but also the t(1) noise in the multi-dimensional spectra. Diffusion measurements with pulsed field gradients characterize lateral movements of lipid and drug molecules in membranes. The theoretical framework for data analysis of MAS diffusion experiments on randomly oriented multilamellar liposomes is presented. PMID- 14745791 TI - Lateral diffusion studied by pulsed field gradient NMR on oriented lipid membranes. AB - This mini-review focuses on the utilization of pulsed magnetic field gradients to measure diffusional motion in systems of macroscopically oriented lipid bilayers. The NMR diffusion technique is proposed to have appreciable potential for future biophysical investigations in the field of membrane biology. Topics such as transport of molecules both across and in the plane of the membrane can be successfully studied, and the formation of lipid domains and their intrinsic dynamics can also be scrutinized. First, a short introduction to the NMR technique is given together with a brief discussion on methods of obtaining a good bilayer orientation. Then, a number of recent results on biophysical/biological membrane systems of great interest is presented, in which some unique conclusions on so-called 'raft membranes' are reached. It is shown for systems with large two-phase areas of liquid disordered and liquid ordered phases that lipid lateral diffusion is faster in the former phase and has a smaller apparent activation energy. Further, on the time-scale of the experiments (50-250 ms), exchange between the two phases is fast in the phospholipid cholesterol-water ternary system, whereas it is slow in the sphingomyelin dioleoylphosphatidylcholine-cholesterol-water quaternary system. PMID- 14745792 TI - Solid-state 2H NMR studies of the effects of cholesterol on the acyl chain dynamics of magnetically aligned phospholipid bilayers. AB - We report the utilization of magnetically aligned phospholipid bilayers (bicelles) to study the effects of cholesterol in phospholipid bilayers for both chain perdeuterated DMPC and partially deuterated alpha-[2,2,3,4,4,6-d(6)] cholesterol using (2)H solid-state NMR spectroscopy. The quadrupolar splittings at 40 degrees C were 25.5 and 37.7 kHz, respectively, for the 2,4-(2)H(eq) and 2,4-(2)H(ax) deuterons when the bilayer normal of the discs was aligned perpendicular to the static magnetic field. The quadrupolar splittings were doubled when Yb(3+) ions were added to flip the bicelles 90 degrees such that the bilayer normal was colinear with the magnetic field. The results suggest that cholesterol is incorporated into the bicelle discs. For chain perdeuterated DMPC d(54), incorporated into DMPC-DHPC bicelle discs, the individual quadrupolar splittings of the methylene and methyl groups doubled on going from the perpendicular to the parallel alignment. Also, the presence of cholesterol increased the overall ordering of the acyl chains of the phospholipids. S(CD) (i) calculations were extracted directly from the (2)H quadrupolar splittings of the chain perdeuterated DMPC. The order parameter, S(CD) (i), calculations clearly indicated an overall degree of ordering of the acyl chains in the presence of cholesterol. We also noted a disordering effect at higher temperatures. This study demonstrates the ease with which (2)H order parameters can be calculated utilizing magnetically aligned phospholipid bilayers when compared with randomly dispersed membrane samples. PMID- 14745793 TI - Properties of polyunsaturated phosphatidylcholine membranes in the presence and absence of cholesterol. AB - Mixtures of cholesterol with phosphatidylcholine species containing the polyunsaturated acyl chains arachidonoyl or docosahexaenoyl were studied by (13)C magic angle spinning (MAS) NMR using both cross-polarization and direct polarization, by (31)P NMR and by differential scanning calorimetry. Several unique features of these systems were observed. The separation of cholesterol in crystalline form occurred at much lower molar fractions than with other forms of phosphatidylcholine. The crystals that were formed were sensitive to the history of the sample. At cholesterol molar fractions below 0.5, they dissolved into the membrane by sequential heating and cooling scans. With higher molar fractions of cholesterol, larger amounts of anhydrous crystals were formed after the first heating. This was accompanied by the formation of non-lamellar phases. The cholesterol crystals that were formed generally were not observed by direct polarization (13)C MAS NMR, even with delay times of 100 s. This suggests that the cholesterol crystals are in a more rigid state in mixtures with these lipids. This is in contrast with the terminal methyl group of the acyl chains that is too mobile to allow cross-polarization using 1 ms contact times. PMID- 14745794 TI - The alphaM1 transmembrane segment of the nicotinic acetylcholine receptor interacts strongly with model membranes. AB - The transmembrane domain of the nicotinic acetylcholine receptor (nAChR) plays a role in the regulation of the activity of this important ligand-gated ion channel. The lipid composition of the host membrane affects conformational equilibria of the nAChR and several classes of inhibitors, most notably anaesthetics, interact directly or indirectly with the four transmembrane M segments, M1-M4, of the nAChR subunits. It has proven difficult to gain insight into structure-function relationships of the M-segments in the context of the entire receptor and the biomembrane environment. However, model membrane systems are well suited to obtain detailed information about protein-lipid interactions. In this solid-state NMR study, we characterized interactions between a synthetic alphaM1 segment of the T. californica nAChR and model membranes of different phosphatidylcholine (PC) lipids. The results indicate that alphaM1 interacts strongly with PC bilayers: the peptide orders the lipid acyl chains and induces the formation of small vesicles, possibly through modification of the lateral pressure profile in the bilayer. The multilamellar vesicle morphology was stabilized by the presence of cholesterol, implying that either the rigidity or the bilayer thickness is a relevant parameter for alphaM1-membrane interactions, which also has been suggested for the entire nAChR. Our results suggest that the model systems are to a certain extent sensitive to peptide-bilayer hydrophobic matching requirements, but that the lipid response to hydrophobic mismatch alone is not the explanation. The effect of alphaM1 on different PC bilayers may indicate that the peptide is conformationally flexible, which in turn would support a membrane-mediated modulation of the conformation of transmembrane segments of the nAChR. PMID- 14745795 TI - Peptide-related alterations of membrane-associated water: deuterium solid-state NMR investigations of phosphatidylcholine membranes at different hydration levels. AB - Deuterated water associated with oriented POPC bilayers was investigated before and after the addition of 2 mol% peptide. Membranes in the presences of antimicrobial-(LAH4), pore-forming- (the segments M2 of influenza A and S4 of the domain I of rat brain sodium channels) or lysine-containing model peptides (LAK1 and LAK3) were investigated by (2)H and proton-decoupled (31)P solid-state NMR. The NMR spectra were recorded as a function of hydration in the range between 15 and 93% relative humidity and of sample composition. In the presence of peptides an increased association of water is observed. A quantitative analysis suggests that the peptide-induced changes in the lipid bilayer packing have a significant effect on membrane-water association. The quadrupolar splittings of (2)H(2)O at a given degree of hydration indicate that the changes of the water deuterium order parameter are specific for the peptide sequence and the lipid composition. PMID- 14745796 TI - Structure determination of aligned samples of membrane proteins by NMR spectroscopy. AB - The paper briefly reviews the process of determining the structures of membrane proteins by NMR spectroscopy of aligned samples, describes the integration of recent developments in the interpretation of spectra of aligned proteins and illustrates the application of these methods to the trans-membrane helical domain of a protein. The emerging methods of interpreting the spectral parameters from aligned samples of isotopically labeled proteins provide opportunities for simultaneously assigning the spectra and determining the structures of the proteins, and also for comparing the results from solid-state NMR experiments on completely aligned samples with those of solution NMR experiments on weakly aligned samples. PMID- 14745798 TI - NMR methods for studying the structure and dynamics of oncogenic and antihistaminic peptides in biomembranes. AB - We present several applications of both wide-line and magic angle spinning (MAS) solid-state NMR of bicelles in which are embedded fragments of a tyrosine kinase receptor or enkephalins. The magnetically orientable bicelle membranes are shown to be of particular interest for studying the functional properties of lipids and proteins in a state that is very close to their natural environment. Quadrupolar, dipolar and chemical shielding interactions can be used to determine minute alterations of internal membrane dynamics and the orientation of peptides with respect to the membrane plane. MAS of bicelles can in turn lead to high resolution proton spectra of hydrated membranes. Using deuterium-proton contrast methods one can then obtain pseudo-high-resolution proton spectra of peptides or proteins embedded in deuterated membranes and determine their atomic 3D structure using quasi-conventional liquid-state NMR methods. PMID- 14745797 TI - Structural studies of apoptosis and ion transport regulatory proteins in membranes. AB - Solid-state NMR spectroscopy is being used to determine the structures of membrane proteins involved in the regulation of apoptosis and ion transport. The Bcl-2 family includes pro- and anti-apoptotic proteins that play a major regulatory role in mitochondrion-dependent apoptosis or programmed cell death. The NMR data obtained for (15)N-labeled anti-apoptotic Bcl-xL in lipid bilayers are consistent with membrane association through insertion of the two central hydrophobic alpha-helices that are also required for channel formation and cytoprotective activity. The FXYD family proteins regulate ion flux across membranes, through interaction with the Na(+), K(+)-ATPase, in tissues that perform fluid and solute transport or that are electrically excitable. We have expressed and purified three FXYD family members, Mat8 (mammary tumor protein), CHIF (channel-inducing factor) and PLM (phospholemman), for structure determination by NMR in lipids. The solid-state NMR spectra of Bcl-2 and FXYD proteins, in uniaxially oriented lipid bilayers, give the first view of their membrane-associated architectures. PMID- 14745799 TI - Temperature dependence and resonance assignment of 13C NMR spectra of selectively and uniformly labeled fusion peptides associated with membranes. AB - HIV-1 and influenza viral fusion peptides are biologically relevant model fusion systems and, in this study, their membrane-associated structures were probed by solid-state NMR (13)C chemical shift measurements. The influenza peptide IFP L2CF3N contained a (13)C carbonyl label at Leu-2 and a (15)N label at Phe-3 while the HIV-1 peptide HFP-UF8L9G10 was uniformly (13)C and (15)N labeled at Phe-8, Leu-9 and Gly-10. The membrane composition of the IFP-L2CF3N sample was POPC-POPG (4:1) and the membrane composition of the HFP-UF8L9G10 sample was a mixture of lipids and cholesterol which approximately reflects the lipid headgroup and cholesterol composition of host cells of the HIV-1 virus. In one-dimensional magic angle spinning spectra, labeled backbone (13)C were selectively observed using a REDOR filter of the (13)C-(15)N dipolar coupling. Backbone chemical shifts were very similar at -50 and 20 degrees C, which suggests that low temperature does not appreciably change the peptide structure. Relative to -50 degrees C, the 20 degrees C spectra had narrower signals with lower integrated intensity, which is consistent with greater motion at the higher temperature. The Leu-2 chemical shift in the IFP-L2CF3N sample correlates with a helical structure at this residue and is consistent with detection of helical structure by other biophysical techniques. Two-dimensional (13)C-(13)C correlation spectra were obtained for the HFP-UF8L9G10 sample and were used to assign the chemical shifts of all of the (13)C labels in the peptide. Secondary shift analysis was consistent with a beta-strand structure over these three residues. The high signal-to-noise ratio of the 2D spectra suggests that membrane-associated fusion peptides with longer sequences of labeled amino acids can also be assigned with 2D and 3D methods. PMID- 14745800 TI - 'Boomerang'-like insertion of a fusogenic peptide in a lipid membrane revealed by solid-state 19F NMR. AB - Solid state (19)F NMR revealed the conformation and alignment of the fusogenic peptide sequence B18 from the sea urchin fertilization protein bindin embedded in flat phospholipid bilayers. Single (19)F labels were introduced into nine distinct positions along the wild-type sequence by substituting each hydrophobic amino acid, one by one, with L-4-fluorophenylglycine. Their anisotropic chemical shifts were measured in uniaxially oriented membrane samples and used as orientational constraints to model the peptide structure in the membrane-bound state. Previous (1)H NMR studies of B18 in 30% TFE and in detergent micelles had shown that the peptide structure consists of two alpha-helical segments that are connected by a flexible hinge. This helix-break-helix motif was confirmed here by the solid-state (19)F NMR data, while no other secondary structure (beta-sheet, 3(10)-helix) was compatible with the set of orientational constraints. For both alpha-helical segments we found that the helical conformation extends all the way to the respective N- and C-termini of the peptide. Analysis of the corresponding tilt and azimuthal rotation angles showed that the N-terminal helix of B18 is immersed obliquely into the bilayer (at a tilt angle tau approximately 54 degrees), whereas the C-terminus is peripherally aligned (tau approximately 91 degrees). The azimuthal orientation of the two segments is consistent with the amphiphilic distribution of side-chains. The observed 'boomerang'-like mode of insertion into the membrane may thus explain how peptide binding leads to lipid dehydration and acyl chain perturbation as a prerequisite for bilayer fusion to occur. PMID- 14745801 TI - Insights into the interactions between a drug and a membrane protein target by fluorine cross-polarization magic angle spinning NMR. AB - The fluorinated anti-psychotic drug trifluoperazine (TFP) has been shown to be a K(+)-competitive inhibitor of gastric H(+)/K(+)-ATPase, a membrane-embedded therapeutic target for peptic ulcer disease. This paper describes how variable contact time (19)F cross-polarization magic angle spinning (VCT-CP/MAS) NMR has been used to probe the inhibitory interactions between TFP and H(+)/K(+)-ATPase in native gastric membranes. The (19)F CP/MAS spectra for TFP in H(+)/K(+)-ATPase enriched (GI) gastric membranes and in control membranes containing less than 5 nmol of the protein indicated that the drug associates with the membranes independently of the presence of H(+)/K(+)-ATPase. The (19)F peak intensities in the VCT-CP/MAS experiment confirmed that TFP undergoes slow dissociation (k(off) < 100 s(-1)) from binding sites in GI membranes, and more rapid dissociation (k(off) < 100 s(-1)) from control membranes. The spectra showed that up to 40% of bound TFP was displaced from GI membranes by 100 mM K(+) and by the K(+) competitive inhibitor TMPIP, but TFP was not displaced from the control membranes. Hence the spectra of TFP in GI membranes represent the drug bound to the K(+)-competitive inhibitory site of H(+)/K(+)-ATPase and to other non specific sites. The affinity of TFP for the K(+)-competitive site (K(D) = 4 mM) was determined from a binding curve of (19)F peak intensity versus TFP concentration after correction for non-specific binding. The K(D) was much higher than the IC(50) for ATPase inhibition (8 microM), which suggests that the substantial non-specific binding of TFP to the membranes contributes to ATPase inhibition. This novel approach to probing ligand binding can be applied to a wide range of membrane-embedded pharmaceutical targets, such as G-protein coupled receptors and ion channels, regardless of the size of the protein or strength of binding. PMID- 14745802 TI - 15N T2' relaxation times of bacteriorhodopsin transmembrane amide nitrogens. AB - 15N T(2)' relaxation times of bacteriorhodopsin (BR) amide nitrogens were determined in the temperature range from 40 to -60 degrees C using a Hahn echo pulse sequence and proton decoupling during the echo and detection times. Using oriented membrane samples, with their bilayer normal parallel to the external magnetic field, the (15)N amide nitrogens belonging to the transmembrane helices could be selected for the analysis. The experiments were performed on purple membrane fragments (in which BR is organized in a 2D crystalline network) and on BR reconstituted into dimyristoylphosphatidylcholine at a 1:150 molar ratio (in which BR is in a freely diffusing monomeric state at 40 degrees C and in an aggregated state at 4 degrees C and below). The results are discussed in terms of helix dynamics, mosaic spread and resolution of the (15)N spectra for the various samples and experimental conditions. PMID- 14745803 TI - Site-directed 13C solid-state NMR studies on membrane proteins: strategy and goals toward revealing conformation and dynamics as illustrated for bacteriorhodopsin labeled with [1-13C]amino acid residues. AB - We have so far demonstrated that well-resolved and site-specifically assigned (13)C peaks as recorded by site-directed NMR study on (13)C-labeled membrane proteins can serve as a convenient probe to reveal their local conformation and dynamics. We attempted here to clarify the extent to which (13)C NMR spectra of (13)C-labeled fully hydrated bacteriorhodopsin (bR) as a typical membrane protein are visible or well resolved in the presence of inherent fluctuation motions with frequency of 10(2)-10(8) Hz, especially at the membrane surfaces. Accordingly, we estimated the relative proportion of (13)C NMR signals from the surface areas with and without peak suppression by the accelerated transverse relaxation effect by surface-bound Mn(2+) ions, which could be effective for residues within 8.7 angstroms of the membrane surface. It turned out that the experimental findings are consistent with the predicted amount of amino acid residues under consideration located within 8.7 angstroms of the surface for [1-(13)C]Val- and Ile-labeled bR and also [3-(13)C]Ala-bR. In contrast, (13)C NMR peaks from such surfaces area are almost completely or partially suppressed for [1-(13)C]Gly-, Ala-, Leu-, Phe- and Trp-labeled bR, as a result of plausible interference of the fluctuation frequency with frequency of magic angle spinning (10(4) Hz). We further assigned several (13)C NMR signals of [1-(13)C] Val-, Trp- and Ile labeled bR on the basis of a variety of site-directed mutants with reference to those of the wild type. Further, we recorded the (13)C NMR of bR in lipid bilayers to search for the optimal conditions to be able to obtain signals with the highest peak intensities and spectral resolution. Backbone dynamics turn out to be essential for recording (13)C NMR spectra so as to escape from motional frequencies of the order of 10(4)-10(5) Hz, either in the direction of accelerated fluctuation or slowed motions in the direction of forming the 2D array. PMID- 14745804 TI - Amyloidosis of Alzheimer's Abeta peptides: solid-state nuclear magnetic resonance, electron paramagnetic resonance, transmission electron microscopy, scanning transmission electron microscopy and atomic force microscopy studies. AB - Aggregation cascade for Alzheimer's amyloid-beta peptides, its relevance to neurotoxicity in the course of Alzheimer's disease and experimental methods useful for these studies are discussed. Details of the solid-phase peptide synthesis and sample preparation procedures for Alzheimer's beta-amyloid fibrils are given. Recent progress in obtaining structural constraints on Abeta-fibrils from solid-state NMR and scanning transmission electron microscopy (STEM) data is discussed. Polymorphism of amyloid fibrils and oligomers of the 'Arctic' mutant of Abeta(1-40) was studied by (1)H,(13)C solid-state NMR, transmission electron microscopy (TEM) and atomic force microscopy (AFM), and a real-time aggregation of different polymorphs of the peptide was observed with the aid of in situ AFM. Recent results on binding of Cu(II) ions and Al-citrate and Al-ATP complexes to amyloid fibrils, as studied by electron paramagnetic resonance (EPR) and solid state (27)Al NMR techniques, are also presented. PMID- 14745805 TI - Structural diversity of amyloid fibril formed in human calcitonin as revealed by site-directed 13C solid-state NMR spectroscopy. AB - Fibril formation in human calcitonin (hCT) from aqueous solution at pH 4.1 was examined and compared with those at pH 3.3 and 7.5 corresponding to three different net charges by means of site-directed (13)C solid-state NMR spectroscopy. Notably, the observed (13)C chemical shifts and lineshapes of the (13)C CP/MAS spectra differed substantially among fibrils prepared at different pHs. It was found that antiparallel beta-sheet structures were formed at pH 7.5 and 4.1 in the central core regions. In the C-terminal region, random coils were formed at both pH 7.5 and 4.1, although the random coil region at pH 4.1 was larger than that at pH 7.5. Fibrillation kinetics analyzed by a two-step autocatalytic reaction mechanism showed that the rate constants k(1) and k(2) for nucleation and maturation reactions of the fibril formation, respectively, were separately determined and the values correlated well with the net positive charges of Lys(18) and His(20) rather than the existence of a negative charge of Asp(15). Further, an attempt was made to assess interatomic distances between amide nitrogen and carbonyl carbon of neighboring chains of (13)C, (15)N-labeled hCT and a model pentapeptide by (13)C REDOR measurements by taking into account its dipolar interaction analyzed by the 3 spin system proposed previously. A unique chain packing of the antiparallel beta-sheets was proposed as a dominant fibril structure, although the possibility of a contribution of chain packing consisting of sliding one or two residues perpendicular to the fibril direction cannot be ruled out. In addition, it appears that the phenyl rings of Phe(16) are aligned on the same side of the beta-sheet and make the beta-sheet stable by forming pi-pi interactions between the beta-strands. PMID- 14745806 TI - Structural role of tyrosine in Bombyx mori silk fibroin, studied by solid-state NMR and molecular mechanics on a model peptide prepared as silk I and II. AB - The influence of the bulky and H-bonding Tyr side-chain on its Ala- and Gly-rich environment in Bombyx mori silk fibroin was examined by (13)C cross-polarization magic angle spinning (CP/MAS), static (2)H and (19)F NMR and molecular mechanics calculations. Model peptides of the type (AG)(15) were synthesized with Tyr in a number of different positions, precipitated under conditions favoring either of the two characteristic protein conformations, and the resulting structures were assigned from their (13)C chemical shifts. Dialysis of native fibroin or the simple (AG)(15) peptide from a 9 M LiBr solution against water produces silk I (the structure of silk before spinning), whereas drying from formic acid yields silk II (fibrous structure after spinning). We found that the introduction one or more Tyr into (AG)(15) can have a dramatic effect not only on the local backbone conformation but also on the long-range intermolecular chain packing in the samples. The antiparallel beta-sheet conformation of silk II is able readily to accommodate a single Tyr residue. Interestingly, the beta-turn conformation of silk I only remains stable when Tyr is positioned near the chain terminus in (AG)(12)YG(AG)(2), but the conformation is driven towards silk II when Tyr is located in the central region of (AG)(7)YG(AG)(7). The role of H-bonding was tested by replacing Tyr with Phe or 4F-Phe, which are no longer compatible with silk I and fully induced a silk II conformation. In the presence of several Tyr residues a mixture of distorted beta-sheet and beta-turn conformations was obtained, regardless of the precipitation conditions. Static (2)H NMR of ring deuterated [3',5'-(2)H(2)]Tyr located in the central region of (AG)(7)YG(AG)(7) showed that the side-chain is immobilized in both silk I and II, which was also observed by static (19)F NMR of the 4F-Phe analogue. To visualize the local packing around the Tyr side-chain, molecular mechanics calculations were performed on a mixture of (AG)(4) and AGAGYGAG, starting from either the beta turn type II or the antiparallel beta-sheet structure. The resulting structures show that the intermolecular chain arrangement is significantly affected by Tyr, thus explaining the long-range packing effects in the semi-crystalline regions of silk fibers compared with the crystalline regions that are devoid of Tyr. PMID- 14745807 TI - Investigation of the dynamics of an elastin-mimetic polypeptide using solid-state NMR. AB - Elastin is the main structural protein that provides elasticity to various tissues and organs in vertebrates. Molecular motions are believed to play a significant role in its elasticity. We have used solid-state NMR spectroscopy to characterize the dynamics of an elastin-mimetic protein as a function of hydration to better understand the origin of elastin elasticity. Poly(Lys-25), [(VPGVG)(4)(VPGKG)](39), has a repeat sequence common to natural elastin. (13)C cross-polarization and direct polarization spectra at various hydration levels indicate that water enhances the protein motion in a non-uniform manner. Below 20% hydration, the backbone motion increases only slightly whereas above 30% hydration, both the backbone and the side-chains undergo large-amplitude motions. The motional amplitudes are extracted from (13)C-(1)H and (1)H-(1)H dipolar couplings using 2D isotropic-anisotropic correlation experiments. The root mean square fluctuation angles are found to be 11-18 degrees in the dry protein and 16 21 degrees in the 20% hydrated protein. Dramatically, the amplitudes increase to near isotropic at 30% hydration. Field-dependent (1)H rotating-frame spin-lattice relaxation times (T(1rho)) indicate that significant motions occur on the microsecond time-scale (1.2-2.3 micros). The large-amplitude and low-frequency motion of poly(Lys-25) at relatively mild hydration indicates that the conformational entropy of the protein in the relaxed state contributes significantly to the elasticity. PMID- 14745808 TI - A solid-state NMR study of the fast and slow dynamics of collagen fibrils at varying hydration levels. AB - We report solid-state NMR investigations of the effect of temperature and hydration on the molecular mobility of collagen isolated from bovine achilles tendon. (13)C cross-polarization magic angle spinning (MAS) experiments were performed on samples at natural abundance, using NMR methods that detect motionally averaged dipolar interactions and chemical shift anisotropies and also slow reorientational processes. Fast motions with correlation times much shorter than 40 micro s scale dipolar couplings and chemical shift anisotropies of the carbon sites in collagen. These motionally averaged anisotropic interactions provide a measure of the amplitudes of the segmental motions expressed by a molecular order parameter. The data reveal that increasing hydration has a much stronger effect on the amplitude of the molecular processes than increasing temperature. In particular, the Cgamma carbons of the hydroxyproline residues exhibit a strong dependence of the amplitude of motion on the hydration level. This could be correlated with the effect of hydration on the hydrogen bonding structure in collagen, for which this residue is known to play a crucial role. The applicability of 1D MAS exchange experiments to investigate motions on the millisecond time-scale is discussed and first results are presented. Slow motions with correlation times of the order of milliseconds have also been detected for hydrated collagen. PMID- 14745809 TI - Improved pulse sequences for pure exchange solid-state NMR spectroscopy. AB - Spin-exchange experiments are useful for improving the resolution and establishment of sequential assignments in solid-state NMR spectra of uniformly (15)N-labeled proteins oriented macroscopically in phospholipid bilayers. To exploit this advantage fully, it is crucial that the diagonal peaks in the two dimensional exchange spectra are suppressed. This may be accomplished using the recent pure-exchange (PUREX) experiments, which, however, suffer from up to a threefold reduction of the cross-peak intensity relative to experiments without diagonal-peak suppression. This loss in sensitivity may severely hamper the applicability for the study of membrane proteins. In this paper, we present a two dimensional exchange experiment (iPUREX) which improves the PUREX sensitivity by 50%. The performance of iPUREX is demonstrated experimentally by proton-mediated (15)N-(15)N spin-exchange experiments for a (15)N-labeled N-acetyl-L-valyl-L leucine dipeptide. The relevance of exchange experiments with diagonal-peak suppression for large, uniformly (15)N-labeled membrane proteins in oriented phospholipid bilayers is demonstrated numerically for the G-protein coupled receptor rhodopsin. PMID- 14745810 TI - Precision 1H-1H distance measurement via 13C NMR signals: utilization of 1H-1H double-quantum dipolar interactions recoupled under magic angle spinning conditions. AB - We applied the POST-C7 DQ-dipolar recoupling pulse sequence to the measurement of (1)H-(1)H distances with high precision. The spectral resolution is enhanced by detecting the (1)H magnetization via (13)C signals. A least-squares fitting of the build-up curve of the transferred magnetization to the exact numerical simulations yielded a (1)H(alpha)-(1)H(beta) distance of 248 +/- 4 pm for fully (13)C-labeled L-valine. This distance agrees with the neutron diffraction study. The negative transferred magnetization clearly indicates that the direct DQ (1)H (1)H dipolar couplings have the largest effect. The signal for the magnetization transfer builds up rapidly by the direct (1)H-(1)H dipolar coupling, and decreases to zero at longer mixing time when the relayed magnetization transfer becomes significant. This large intensity change of the signal leads to the high precision in the distance measurement. We inspected factors that limit the effective bandwidth of the POST-C7 recoupling for the (1)H and (13)C homonuclear spin systems. The spin interactions at times shorter than the cycle time of the C7 sequence were also evaluated to measure the distances. The carbon-detected 2D (1)H DQ mixing experiment was demonstrated for the measurement of multiple (1)H (1)H distances. PMID- 14745811 TI - Non-empirical calculations of NMR indirect carbon-carbon coupling constants. Part 6: propellanes. AB - A full set of carbon-carbon coupling constants have been calculated at the SOPPA level in the series of six most representative propellanes. Special attention was focused on spin-spin couplings involving both bridgehead carbons, and these data were rationalized in terms of the multipath coupling mechanism and hybridization effects. Many unknown couplings in the propellane frameworks were predicted with high reliability. PMID- 14745812 TI - Identifying residues in natural organic matter through spectral prediction and pattern matching of 2D NMR datasets. AB - This paper describes procedures for the generation of 2D NMR databases containing spectra predicted from chemical structures. These databases allow flexible searching via chemical structure, substructure or similarity of structure as well as spectral features. In this paper we use the biopolymer lignin as an example. Lignin is an important and relatively recalcitrant structural biopolymer present in the majority of plant biomass. We demonstrate how an accurate 2D NMR database of approximately 600 2D spectra of lignin fragments can be easily constructed, in approximately 2 days, and then subsequently show how some of these fragments can be identified in soil extracts through the use of various search tools and pattern recognition techniques. We demonstrate that once identified in one sample, similar residues are easily determined in other soil extracts. In theory, such an approach can be used for the analysis of any organic mixtures. PMID- 14745813 TI - Experimental and theoretical NMR study of selected oxocarboxylic acid oximes. AB - 1H and 13C NMR spectra of the oxocarboxylic acid oximes 2-hydroxyiminopropanoic acid (1), 2-(4-methylthiazol-2-yl)-2-(hydroxyimino)acetic acid (2) and 2-cyano-2 (hydroxyimino)acetic acid (3) were measured in DMSO-d6, D2O and acetone-d6 solutions. The data indicate the presence of hydrogen bonding in 1 and 2 and a strong electron-withdrawing effect due to the cyano group in 3. The effect of intra- and intermolecular hydrogen bonding on the hydrogen and carbon chemical shifts in these molecules was studied theoretically. Total energy calculations of the stability of various hydrogen-bonded species, in addition to equilibrium parameters and chemical shifts, were calculated using ab initio methods (RHF, MP2) and density functional theory (B3LYP), implemented in the Gaussian 98 software package. The gauge-including atomic orbital (GIAO) method was used to predict magnetic shielding constants. Chemical shift calculations for the most stable species agree fairly well with the observed data, especially for the hydroxyl protons. Substituents adjacent to the alpha-carbon show some influence of the oximic and carboxyl groups on the 13C chemical shifts, as expected for groups with different polar and anisotropic character. PMID- 14745814 TI - The aluminum ordering in aluminosilicates: a dipolar 27Al NMR spectroscopy study. AB - The spatial ordering of aluminum atoms in CsAl(SiO3)2 and 3Al2O3.2SiO2 was probed by 27Al dipolar solid-state NMR spectroscopy. The 27Al response to a Hahn spin echo pulse sequence in a series of aluminum-containing model crystalline compounds demonstrates that quantitative 27Al homonuclear dipolar second moments can be obtained to within +/-20% of the theoretical values, if evaluation of the spin-echo response curve is limited to short evolution periods (2t1 < or = 0.10 ms). Additionally, selective excitation of the central transition m = 1/2 --> 1/2 is necessary in order to ensure quantitative results. Restriction of spin exchange affecting the dephasing of the magnetization may decelerate the spin echo decay at longer evolution periods. Considering these restraints, the method was used to probe the spatial distribution of aluminum atoms among the tetrahedral sites in two aluminosilicate materials. Experimental 27Al spin-echo response data for the aluminosilicates CsAl(SiO3)2 (synthetic pollucite) and 3Al2O3.2SiO2 (mullite) are compared with theoretical data based on (I) various degrees of aluminum-oxygen-aluminum bond formation among tetrahedrally coordinated aluminum atoms (Al(T(d) )-O-Al(T(d) )) and (II) the maximum avoidance of Al(T(d) )-O-Al(T(d) ) bonding. Analysis of the second moment values and resulting echo decay responses suggests that partial suppression of spin exchange among aluminum atoms in crystallographically distinct sites may contribute to the 27Al spin echo decay in 3Al2O3.2SiO2, thus complicating quantitative analysis of the data. Silicon-29 and aluminum-27 magic angle spinning (MAS) NMR spectra of 3Al2O3.2SiO2 are consistent with those previously reported. The experimental 27Al spin-echo response behavior of CsAl(SiO3)2 differs from the theoretical response behavior based on the maximum avoidance of Al-O-Al bonding between tetrahedral aluminum sites in CsAl(SiO3)2. A single unresolved resonance is observed in both the silicon-29 and aluminum-27 MAS spectra of CsAl(SiO3)2. PMID- 14745815 TI - Dynamic NMR and ab initio studies of exchange between rotamers of derivatives of octahydrofuro[3,4-f]isoquinoline-7(1H)-carboxylate and tetrahydro-2,5,6(1H) isoquinolinetricarboxylate. AB - The 1H and 13C NMR spectra of methyl-8-(2-furyl)-5-methyl-1,3-dioxo 3,3a,4,6,8,9,9a,9b-octahydrofuro[3,4-f]isoquinoline-7(1H)-carboxylate (1) and trimethyl 8-methyl-3-phenyl-3,4,4a,7-tetrahydro-2,5,6(1H) isoquinolinetricarboxylate (2) at room temperature displayed doubling of the majority of signals, suggesting the presence of two rotational conformers (rotamers) in a ratio approximately 1:1.2 (in a mixture of CDCl3 and C6D6), approximately 1:1 (in CD2Cl2) and approximately 1:1.4 (in C6D6). On the basis of the temperature-dependent 1H NMR spectra of 1 and 2, the barrier to interconversion of the rotamers was calculated to be approximately 16 kcal mol( 1). The average chemical shifts and spin-spin coupling constants were analyzed for the resolution-enhanced 300 MHz 1H NMR spectrum of 1 at 333 K in CDCl3 solution. From analysis of the spin-spin coupling constants, it is proposed that the nitrogen-containing ring is in a chair conformation with C-2-H equatorial. Low- and room-temperature NOESY experiments in conjunction with theoretical ab initio calculations supported the hypothesis that the two rotamers interchange via rotation about the C-N bond. Spectral assignments of all proton and carbon resonances were made on the basis of one- and two-dimensional NMR experiments (DEPT, DQCOSY, NOESY, HETCOR and gHMBC). PMID- 14745816 TI - Application of the TORO technique of 1H NMR to the structural analysis of cyclic peptide isomers having a slightly distorted symmetry from C2. AB - The extended TORO technique was applied to the structural analysis of endo-D-Tyr gramicidin S, cyclo(-Val-Orn-Leu-D-Phe-D-Tyr-Pro-Val-Orn-Leu-D-Phe-Pro-), which has a slightly distorted symmetry from C2, by the insertion of D-Tyr and equivalent alpha-proton chemical shifts in the 1H NMR spectrum. All NMR signals of the two dominant isomers of this antibiotic with trans-trans prolines were determined by using the extended TORO technique with TOCSY and ROESY spectra. This technique is generally applicable for distinguishing overlapped signals of alpha- and amide protons from the main chains of peptides. PMID- 14745817 TI - HMBC-RELAY: a combined technique for the differentiation of simultaneously detected 2J(C,H) and (n)J(C,H) connectivities. AB - We present a new pulse sequence that detects simultaneously (n)J(C,H) and 2J(C,H) connectivities. The corresponding coherences are created along independent pathways and therefore can be separated into two different subspectra. One spectrum is to show all (n)J(C,H) connectivities and the other is to show exclusively 2J(C,H) connectivities. In contrast to the previously published 2J/(n)J experiment, this sequence detects the 2J(C,H) connectivities via a C,H,H RELAY pathway leading to an intensification of the 2J(C,H) signals. Strictly, the 2J(C,H) spectrum does not show 2J(C,H) but 3J(H,H) coupling interactions within 13CH(k)-12CH(l) fragments. Therefore, 2J(C,H) signals can appear even if the corresponding 2J(C,H) coupling constant is zero. PMID- 14745818 TI - Evaluation of protein 15N relaxation times by inverse Laplace transformation. AB - Relaxation times (T1, T2, T1rho) are usually evaluated from exponential decay data by least-squares fitting methods. For this procedure, the integrals or amplitudes of signals must be determined, which can be laborious with large data sets. Moreover, the fitting requires a priori knowledge of the number of exponential components responsible for the decay. We have adapted inverse Laplace transformation (ILT) for the analysis of relaxation data. Exponential components are resolved with ILT to reciprocal space on their corresponding relaxation rate values. The ILT approach was applied to 3D linewidth-resolved 15N HSQC experiments to evaluate 15N T1 and T2 relaxation times of ubiquitin. The resulting spectrum is a true 3D spectrum, where the signals are separated by their 1H and 15N chemical shifts (HSQC correlations) and by their relaxation rate values (R1 or R2). From this spectrum, the relaxation times can be obtained directly with a simple peak-picking procedure. PMID- 14745819 TI - Orientations of the principal components of electric field gradients and internal motions in dihydrogen ligands from the 2H T1 NMR relaxation data in solution. AB - The deuterium spin-lattice relaxation times in (D2) ligands of W, Ru and Os complexes are reviewed and analyzed in terms of the fast internal (D2) motions: free rotation, librations and 180 degrees jumps. The analysis was performed using quadrupolar coupling constant (DQCC) parameters taken from the solid-state 2H NMR spectra and density function theory calculations. It is shown that the calculated DQCC values can be corrected for further use in interpretations of deuterium relaxation times for Ru and Os dihydrogen complexes. The resulting data led to a criterion for using the relaxation data to distinguish fast-spinning dihydrogen ligands. It is shown that the principal components of electric field gradient tensors at D in the dihydrogen ligands are oriented closer to M-D directions. PMID- 14745820 TI - Evidence for spin diffusion in a H,H-NOESY study of imidazolium tetrafluoroborate ionic liquids. AB - The ionic liquids 1-ethyl-3-methylimidazolium tetrafluoroborate ([EMIM][BF4]) and 1-methyl-3-propylimidazolium tetrafluoroborate ([PMIM][BF4]) were studied by H,H NOESY NMR using a cross-relaxation matrix analysis. Cross-peak intensities are seen to increase with increasing mixing time. Experimental and theoretical hydrogen-hydrogen distances are in agreement at short mixing times (50 ms). Mixing times longer than 50 ms result in an increasing contribution of spin diffusion that produces unrealistically short hydrogen-hydrogen distances. Gas phase ab initio molecular structures are obtained using Hartree-Fock (HF) and density functional theory (B3LYP) methods at the 6311 + G(2d,p) basis set level. The hydrogen-hydrogen distances obtained from the theoretical structures are in reasonable agreement with those calculated from the cross-relaxation matrices. PMID- 14745821 TI - Quantitative estimation of the degree of derivatization: an alternative methodology using 1-D and 2-D NMR experiments. AB - The precise estimation of the degree of derivatization of functional groups in polymers is important for determining their macroscopic properties. In this work, the quantitative estimation of the extent of esterification of novolac copolymers with di-tert-butyl dicarbonate was studied. Although the extent of esterification has been calculated previously by quantifying the signals from FT-IR and UV-Vis spectroscopy, these were restricted to monitoring the progress of the derivatization process. The 13C NMR signal intensities from the inverse-gated 1H decoupled NMR spectrum have been used recently for the quantitative estimation of the degree of esterification of polymers. An alternative methodology has been suggested by us based on the fully relaxed 1H chemical shift intensities. However, since the proton signals of novolac resins are generally broad and overlapping, the proton decoupled 13C NMR spectrum was used to identify the 1H NMR signals using the 2-D HSQC technique. A TOCSY experiment was also performed to confirm further the 1H NMR signal assignments and, finally, the deconvoluted 1H NMR spectrum was used for the calculation of the extent of derivatization. PMID- 14745822 TI - Complete assignment of the 1H, 13C, 15N and 19F NMR spectra of nine DL phenylalcoholamide anticonvulsants. AB - The 1H, 13C, 15N and 19F NMR spectra of nine DL-phenylalcoholamides bearing fluorine and chlorine as substituents of the phenyl ring are reported. All of them are active as anticonvulsants in pentylenetetrazole-induced seizures. PMID- 14745823 TI - The effect of mutations altering biogenic amine metabolism in Drosophila on viability and the response to environmental stresses. AB - Dopamine (DA) content, tyrosine decarboxylase (TDC) activity and survival were studied under normal and environmental stress conditions in the ste and e strains carrying ebony mutation increasing DA level and the octopamineless strain Tbetah(nM18) of Drosophila melanogaster. Wild-type strains Canton S and Oregon R, and strain p845 from which Tbetah(nM18) strain was derived were used as controls. Sexual dimorphism of TDC activity, DA content, and survival in flies of all D. melanogaster strains under study was found. Tbetah(nM18) mutation sharply reduced TDC activity in females, while ebony had no such effect. DA content and survival under heat stress in Tbetah(nM18) flies did not differ from those in the wild type. ste and e flies had drastically increased DA content under normal conditions, dramatically decreased survival under heat stress, but increased survival under starvation. DA content and survival under heat stress were also studied in the reciprocal hybrids (males) F(1) of the cross D. virilis strains 101 (wild type) and 147 with X-linked mutation, which significantly increases DA content. 147x101 males had a considerably higher DA content and lower survival than 101x147 ones. Individuals of all D. melanogaster strains under study developed the stress reaction, as judged by changes in TDC activity and DA levels. The role of biogenic amines in the stress reaction development and adaptation to environmental stresses in Drosophila is discussed. Arch. Insect Biochem. Physiol. 55:55-67, 2004. PMID- 14745824 TI - Morphological and molecular effects of 20-hydroxyecdysone and its agonist tebufenozide on CF-203, a midgut-derived cell line from the spruce budworm, Choristoneura fumiferana. AB - The morphological and molecular responses of a midgut-derived cell line of the spruce budworm, Choristoneura fumiferana, to 20-hydroxyecdysone (20E) and the nonsteroidal ecdysone agonist, tebufenozide (RH-5992), were investigated. The cells responded to these compounds by clumping, generating filamentous extensions, increased mortality and expression of the transcription factor, Choristoneura hormone receptor 3 (CHR3). This cell line can be used as a model system to study the mode of action of ecdysone and its agonists. With subsequent passaging in ecdysteroid-containing medium, the degree of clumping increased and the clumping could not be reversed by subculturing in ecdysteroid-free medium. Cell numbers of the adapted cell lines in 20E and RH-5992 containing media were not significantly decreased, compared to the control, but both cell lines accumulated less (14)C-labeled RH-5992 and lost the capability of expressing CHR3 in response to these compounds. Taken together, the cell lines appeared to develop a mechanism to adapt to the toxic effects of these compounds. Arch. Insect Biochem. Physiol. 55:68-78, 2004. PMID- 14745825 TI - Role of antioxidant defense during different stages of preadult life cycle in European corn borer (Ostrinia nubilalis, Hubn.): Diapause and metamorphosis. AB - Antioxidant enzymes, total glutathione (GSH), and ascorbic acid (ASA) were determined in whole body homogenates of nondiapausing larvae, diapausing larvae during the diapausing period (October, December, and February), and in pupae emerged from both diapausing and nondiapausing larvae of the European corn borer (Ostrinia nubilalis, Hubn., Lepidoptera: Pyralidae). The activities of catalase, selenium nondependent glutathione peroxidase (GPx), and glutathione-S-transferase (GST), as well as the content of GSH and ASA, were found to vary throughout the larval diapause. Compared to diapausing larvae, nondiapausing larvae were higher in levels of catalase, GPx, GST, and dehydroascorbate reductase (DHAR) activity. GSH content was also increased. However, nondiapausing larvae contained less ASA than diapausing ones. Pupae had higher GPx and GST activity and an increased ASA content compared to larvae. The pupae emerged from nondiapausing larvae had higher GST, glutathione reductase (GR), and DHAR activities, but lower GPx activity and ASA content than those emerged from diapausing larvae. Correlation analysis revealed differences in the way the antioxidant level is equilibrated for a particular stage and developmental pattern. The results suggest that cellular antioxidants are involved in both the protection of cells and the regulation of redox levels during the pre-adult stages of Ostrinia nubilalis. Arch. Insect Biochem. Physiol. 55:79-89, 2004. PMID- 14745826 TI - Changes in leucine transport activity in Chironomus riparius larvae after short term exposure to potassium dichromate and fenitrothion. AB - The effect of sublethal concentrations of potassium dichromate and fenitrothion on sodium-leucine cotransport in brush border membrane vesicles from Chironomus riparius larvae has been investigated. Exposure to potassium dichromate and fenitrothion caused a dose- and time-dependent inhibition of leucine uptake. Transport inhibition is easily detectable at doses 100-fold lower than LD50. Kinetic experiments showed that inhibition was mainly caused by a decrease of the Vmax (680 +/- 53 vs. 382 +/- 23 and 555 +/- 27 nmol/15s/mg protein in control and exposed larvae to K2Cr2O7 and fenitrothion, respectively). Inhibition is possibly related to a variation of sodium ions permeability as evidenced by increased membrane lipid peroxidation. Appropriate control experiments ruled out that the observed differences could be due to changes in general features of membrane preparations. Transport inhibition observed in larvae exposed to potassium dichromate was accompanied by changes in ascorbate peroxidase and dehydroascorbate reductase activities, whereas those exposed to fenitrothion displayed an increase in transaminase activity. The possible value of leucine uptake as biochemical biomarker is briefly discussed. Arch. Insect Biochem. Physiol. 55:90-101, 2004. PMID- 14745828 TI - Expansion of the genetic code in yeast: making life more complex. AB - Proteins account for the catalytic and structural versatility displayed by all cells, yet they are assembled from a set of only 20 common amino acids. With few exceptions, only 61 nucleotide triplets also direct incorporation of these amino acids. Endeavors to expand the genetic code recently progressed to nucleus containing cells, after Chin et al.1 transferred Escherichia coli genes for a mutant tyrosine-adaptor molecule and its synthetase into Saccharomyces cerevisiae. Transformed yeast cells were produced that exhibit efficient site specific incorporation of non-biotic amino acids into proteins. This makes it likely that code complexity can be elevated experimentally in mammals. PMID- 14745829 TI - Deciphering the swordtail's tale: a molecular and evolutionary quest. AB - The power of sexual selection to influence the evolution of morphological traits was first proposed more than 130 years ago by Darwin. Though long a controversial idea, it has been documented in recent decades for a host of animal species. Yet few of the established sexually selected features have been explored at the level of their genetic or molecular foundations. In a recent report, Zauner et al.1 describe some of the molecular features associated with one of the best characterized of sexually selected traits, the male-specific tail "sword" seen in certain species of the fish genus Xiphophorus. Zauner et al. find that the msxC gene, a gene previously implicated in fin development from work in zebrafish, is dramatically and specifically upregulated in the development of the ventral caudal fin rays, which give rise to the sword, in males. The results provide the first molecular insight into the development of this sexually selected trait while prompting new questions about the structure of the entire genetic network that underlies this trait. To fully understand the molecular-genetic and evolutionary history of this network, however, it will be essential to determine whether sword-development is a basal or derived trait in Xiphophorus. PMID- 14745830 TI - Sex determination: insights from the chicken. AB - Not all vertebrates share the familiar system of XX:XY sex determination seen in mammals. In the chicken and other birds, sex is determined by a ZZ:ZW sex chromosome system. Gonadal development in the chicken has provided insights into the molecular genetics of vertebrate sex determination and how it has evolved. Such comparative studies show that vertebrate sex-determining pathways comprise both conserved and divergent elements. The chicken embryo resembles lower vertebrates in that estrogens play a central role in gonadal sex differentiation. However, several genes shown to be critical for mammalian sex determination are also expressed in the chicken, but their expression patterns differ, indicating functional plasticity. While the genetic trigger for sex determination in birds remains unknown, some promising candidate genes have recently emerged. The Z linked gene, DMRT1, supports the Z-dosage model of avian sex determination. Two novel W-linked genes, ASW and FET1, represent candidate female determinants. PMID- 14745831 TI - The many colours of chromodomains. AB - Local differences in chromatin organisation may profoundly affect the activity of eukaryotic genomes. Regulation at the level of DNA packaging requires the targeting of structural proteins and histone-modifying enzymes to specific sites and their stable or dynamic interaction with the nucleosomal fiber. The "chromodomain", a domain shared by many regulators of chromatin structure, has long been suspected to serve as a module mediating chromatin interactions in a variety of different protein contexts. However, recent functional analyses of a number of different chromodomains revealed an unexpected diversity of interaction targets, including histones, DNA and even RNA. The chromodomains of today seem to have evolved from a common ancestral fold to fulfill various functions in different molecular contexts. Combining information gained from recent functional and structural studies of chromodomains with a bioinformatic classification of their structure could lead to the definition of sequence motifs with predictive quality for chromodomain function. PMID- 14745832 TI - LIMK1 and CLIP-115: linking cytoskeletal defects to Williams syndrome. AB - Williams Syndrome is a developmental disorder that is characterized by cardiovascular problems, particular facial features and several typical behavioral and neurological abnormalities. In Williams Syndrome patients, a heterozygous deletion is present of a region on chromosome 7q11.23 (the Williams Syndrome critical region), which spans approximately 20 genes. Two of these genes encode proteins that regulate dynamic aspects of the cytoskeleton of the cell, either via the actin filament system (LIM kinase 1, or LIMK1), or through the microtubule network (cytoplasmic linker protein of 115 kDa, or CLIP-115). The recent findings that knockout mice lacking LIMK1 or CLIP-115 have distinct neurological and behavioural phenotypes, indicates that cytoskeletal defects might play a role in the development of neurological symptoms in Williams Syndrome patients. In this review, we discuss the properties of LIMK and CLIP family proteins, their function in the regulation of the actin and microtubule cytoskeletal systems, respectively, and the relationship with neurodevelopmental aspects of Williams Syndrome. PMID- 14745833 TI - Post-replication repair in DT40 cells: translesion polymerases versus recombinases. AB - Replication forks inevitably stall at damaged DNA in every cell cycle. The ability to overcome DNA lesions is an essential feature of the replication machinery. A variety of specialized polymerases have recently been discovered, which enable cells to replicate past various forms of damage by a process termed translesion synthesis. Alternatively, homologous recombination can be used to restart DNA replication across the lesion. Genetic and biochemical studies have shed light on the impact of these two post-replication repair pathways in bacteria and yeast. In vertebrates, however, a genetic approach to study post replication repair has been compromised because many of the genes involved appear to be essential for embryonic development. We have taken advantage of the chicken cell line DT40 to perform a genetic analysis of translesion synthesis and homologous recombination and to characterize genetic interactions between these two pathways in vertebrates. In this article, we aim to summarize our current understanding of post-replication repair in DT40 in the perspective of bacterial, yeast and mammalian genetics. PMID- 14745834 TI - How mammalian sex chromosomes acquired their peculiar gene content. AB - It has become increasingly evident that gene content of the sex chromosomes is markedly different from that of the autosomes. Both sex chromosomes appear enriched for genes related to sexual differentiation and reproduction; but curiously, the human X chromosome also seems to bear a preponderance of genes linked to brain and muscle functions. In this review, we will synthesize several evolutionary theories that may account for this nonrandom assortment of genes on the sex chromosomes, including 1) asexual degeneration, 2) sexual antagonism, 3) constant selection, and 4) hemizygous exposure. Additionally, we will speculate on how the evolution of sex-chromosome gene content might have impacted on the phenotypic evolution of mammals and particularly humans. Our discussion will focus on the mammalian sex chromosomes, but will cross reference other species where appropriate. PMID- 14745835 TI - The use of animal models in the study of complex disease: all else is never equal or why do so many human studies fail to replicate animal findings? AB - The study of the genetics of complex human disease has met with limited success. Many findings with candidate genes fail to replicate despite seemingly overwhelming physiological data implicating the genes. In contrast, animal model studies of the same genes and disease models usually have more consistent results. We propose that one important reason for this is the ability to control genetic background in animal studies. The fact that controlling genetic background can produce more consistent results suggests that the failure to replicate human findings in the same diseases is due to variation in interacting genes. Hence, the contrasting nature of the findings from the different study designs indicates the importance of non-additive genetic effects on human disease. We discuss these issues and some methodological approaches that can detect multilocus effects, using hypertension as a model disease. This article contains supplementary material, which may be viewed at the BioEssays website at http://www.interscience.wiley.com/jpages/0265-9247/suppmat/index.html. PMID- 14745836 TI - Recombination in HIV and the evolution of drug resistance: for better or for worse? AB - The rapid evolution of drug resistance remains a major obstacle for HIV therapy. The capacity of the virus for recombination is widely believed to facilitate the evolution of drug resistance. Here, we challenge this intuitive view. We develop a population genetic model of HIV replication that incorporates the processes of mutation, cellular superinfection, and recombination. We show that cellular superinfection increases the abundance of low fitness viruses at the expense of the fittest strains due to the mixing of viral proteins during virion assembly. Moreover, we argue that whether recombination facilitates the evolution of drug resistance depends critically on how resistance mutations interact to determine viral fitness. Contrary to the commonly held belief, we find that, under the most plausible biological assumptions, recombination is expected to slow down the rate of evolution of multi-drug-resistant virus during therapy. PMID- 14745837 TI - A cluster translocation model may explain the collinearity of Hox gene expressions. AB - A model is proposed that deals with the observed collinearities (spatial, temporal and quantitative) of Hox gene expression during pattern formation along the primary and secondary axes of vertebrates. In particular, in the proximodistal axis of the developing limb, it is assumed that a morphogen gradient is laid down with its source at the distal tip of the bud. The extracellular signals in every cell of the morphogenetic field are transduced and uniformly amplified so that molecules are produced in the nucleus with appropriate physicochemical properties. These molecules can exert a concentration dependent force on the Hox cluster. It is assumed that, before activation, the Hox cluster is packaged as an elongated rigid body inside the chromatin and is covered by a coat that prevents the transcription factors reaching the genes of the cluster. The transcription factors are confined to the interchromatin domain and their density decreases with their distance from the chromatin surface. A gradual increase in the extracellular morphogen concentration causes a corresponding increase in the number of the nuclear molecules and the resulting bigger force pushes the Hox cluster toward the interchromatin domain. The step-by step translocations of the Hox cluster initiate the consecutive exposure of genes to their transcription factors. The model explains how gene activation is triggered and it describes spatial, temporal and quantitative collinearities at the initial stages of gene expression. Some recent experiments of Hox deletions and duplications are accounted for by the model. PMID- 14745838 TI - Analyzing protein-protein interactions in cell membranes. AB - Interactions among membrane proteins regulate numerous cellular processes, including cell growth, cell differentiation and apoptosis. We need to understand which proteins interact, where they interact and to which extent they interact. This article describes a set of novel approaches to measure, on the surface of living cells, the number of clusters of proteins, the number of proteins per cluster, the number of clusters or membrane domains that contain pairs of interacting proteins and the fraction of one protein species that interacts with another protein within these domains. These data can then be interpreted in terms of the function of the protein-protein interactions. PMID- 14745839 TI - Interview with Gerald M. Edelman: part I. PMID- 14745840 TI - Significance of the detection of esters of p-hydroxybenzoic acid (parabens) in human breast tumours. AB - This issue of Journal of Applied Toxicology publishes the paper Concentrations of Parabens in Human Breast Tumours by Darbre et al. (2004), which reports that esters of p-hydroxybenzoic acid (parabens) can be detected in samples of tissue from human breast tumours. Breast tumour samples were supplied from 20 patients, in collaboration with the Edinburgh Breast Unit Research Group, and analysed by high-pressure liquid chromatography and tandem mass spectrometry. The parabens are used as antimicrobial preservatives in underarm deodorants and antiperspirants and in a wide range of other consumer products. The parabens also have inherent oestrogenic and other hormone related activity (increased progesterone receptor gene expression). As oestrogen is a major aetiological factor in the growth and development of the majority of human breast cancers, it has been previously suggested by Darbre that parabens and other chemicals in underarm cosmetics may contribute to the rising incidence of breast cancer. The significance of the finding of parabens in tumour samples is discussed here in terms of 1). Darbre et al's study design, 2). what can be inferred from this type of data (and what can not, such as the cause of these tumours), 3). the toxicology of these compounds and 4). the limitations of the existing toxicology database and the need to consider data that is appropriate to human exposures. PMID- 14745841 TI - Concentrations of parabens in human breast tumours. AB - Parabens are used as preservatives in many thousands of cosmetic, food and pharmaceutical products to which the human population is exposed. Although recent reports of the oestrogenic properties of parabens have challenged current concepts of their toxicity in these consumer products, the question remains as to whether any of the parabens can accumulate intact in the body from the long-term, low-dose levels to which humans are exposed. Initial studies reported here show that parabens can be extracted from human breast tissue and detected by thin layer chromatography. More detailed studies enabled identification and measurement of mean concentrations of individual parabens in samples of 20 human breast tumours by high-pressure liquid chromatography followed by tandem mass spectrometry. The mean concentration of parabens in these 20 human breast tumours was found to be 20.6 +/- 4.2 ng x g(-1) tissue. Comparison of individual parabens showed that methylparaben was present at the highest level (with a mean value of 12.8 +/- 2.2 ng x g(-1) tissue) and represents 62% of the total paraben recovered in the extractions. These studies demonstrate that parabens can be found intact in the human breast and this should open the way technically for more detailed information to be obtained on body burdens of parabens and in particular whether body burdens are different in cancer from those in normal tissues. PMID- 14745842 TI - Microarray profile analysis of toxic cocaine-induced alterations in the expression of mouse brain gene sequences: a possible 'protective' effect of buprenorphine. AB - Using a mouse brain cDNA microarray consisting of 2688 gene sequences, which include unknown sequences with the empirical possibility of expression in the brain, the effects of repeated toxic doses of intraperitoneal (i.p.) cocaine (40 mg x kg(-1), 4 days) on the expression pro fi le of the cerebral genes were investigated. The modifications in this pro file caused by buprenorphine (BUP) (0.25 mg x kg(-1) i.p., 4 days), a protective drug against cocaine, were also examined. In the cocaine group, the expression levels reached the recommended increased levels (>or=2 times the control value in the saline-treated control group) in 24.0% of the genes but were equal to or less than the recommended attenuation levels (0.5 and <2 times the control value). Although statistically significant modifications in the expression of cocaine- or BUP-related brain-region-specific genes were not proved using whole cerebrums, including many unknown genes, our results suggest that the expression of genes related to neuronal cell damage, including non-peculiar genes related to the damage accompanying convulsive seizures and malignant tumors, were normalized and the genes related to the protection of neural cells were induced by BUP. PMID- 14745843 TI - Protective effect of alpha-lipoic acid against chloroquine-induced hepatotoxicity in rats. AB - Oral administration of a-lipoic acid, a metavitamin, was investigated for its possible hepatoprotective effect in Wistar rats against chloroquine-induced toxicity. Rats were treated orally with alpha-lipoic acid (10, 30 and 100 mg x kg(-1) day(-1)) for 7 days before a single oral administration of chloroquine (970 mg x kg(-1) day(-1)) and alpha-lipoic acid treatment was continued for three more days. The increased level of serum enzymes (aspartate transaminase, alanine transaminase and alkaline phosphatase), bilirubin, lipids and plasma thiobarbituric acid-reactive substances (TBARS) and hydroperoxides observed in rats treated with chloroquine were very much reduced in rats treated with alpha lipoic acid plus chloroquine. A significant decrease in plasma antioxidants such as reduced glutathione (GSH), vitamin C and vitamin E were observed in chloroquine-treated rats when compared with control rats. Administration of alpha lipoic acid significantly improved the levels of plasma antioxidants GSH, vitamin C and vitamin E in chloroquine-treated rats. In the case of 100 mg x kg(-1) day( 1) the effect was highly significant compared with the other doses (10 and 30 mg x kg(-1) day(-1)). The results of the study revealed that alpha-lipoic acid could offer protection against chloroquine-induced hepatotoxicity. alpha-Lipoic acid had a better protective effect when compared with silymarin, a reference drug. PMID- 14745844 TI - Role of caffeic acid phenethyl ester, an active component of propolis, against cisplatin-induced nephrotoxicity in rats. AB - We have investigated the effect of caffeic acid phenethyl ester (CAPE) on cisplatin-induced nephrotoxicity in rats. Administration of a single dose of cisplatin resulted in the elevation of blood urea nitrogen and creatinine in serum, as well as nitric oxide in kidney tissue of rats. Cisplatin also caused reduction of catalase (P < 0.0001), superoxide dismutase (P = 0.149) and glutathrone peroxidase (P < 0.0001) activities in kidney tissue. Although cisplatin caused elevation in malondialdehyde levels and myeloperoxidase activities in kidney tissue, they were not statistically significant. Caffeic acid phenethyl ester was found to be protective against cisplatin-induced antioxidant enzyme reductions. Treatment with free-radical scavenger CAPE attenuated the increase in plasma blood urea nitrogen and kidney nitric oxide levels, and showed histopathological protection against cisplatin-induced acute renal failure. Extensive epithelial cell vacuolization, swelling, desquamation and necrosis were observed in the kidney of the cisplatin-treated rat. There were also larger tubular lumens in cisplatin-treated rats than those of the control and the CAPE groups. Caffeic acid phenethyl ester caused a marked reduction in the extent of tubular damage. It is concluded that administration of cisplatin imposes an oxidative stress to renal tissue and CAPE confers protection against the oxidative damage associated with cisplatin. This mechanism may be attributed to its free-oxygen-radical scavenging activity. PMID- 14745845 TI - Examination of changes in connective tissue macromolecular components of large white pig skin following application of Lewisite vapour. AB - The aim of this study was to provide information about the degradative processes that occur in major connective tissue components in skin following exposure of large white pigs to Lewisite vapour. Of particular interest were alterations in glycoproteins, which are known to mediate dermo-epidermal attachment (laminin and type IV collagen) and the main collagen found in the dermis (type III collagen). The immunostaining of transfer blots from skin extracts run on sodium dodecyl sulphate polyacrylamide gel electrophoresis gels revealed no evidence of cross linking of laminin or of type III or IV collagen. However, there was evidence of a very considerable degradation of laminin and, to a lesser extent, of type IV collagen. Type III collagen did not appear to be degraded in skin exposed to Lewisite. These degradative processes appeared to be more severe than found in previous studies in Yucatan mini-pigs percutaneously exposed to sulphur mustard, in which only laminin was found to undergo partial cleavage rather than wholesale degradation. The results suggest that damage to macromolecular components in the sub-epidermal basement membrane in skin which mediate dermo-epidermal separation processes may be a common feature in the mechanism of action of vesicating agents such as Lewisite and sulphur mustard. It is of interest that the damage to laminin in this study appeared to be more severe than that previously found for sulphur mustard. This suggests that skin can suffer substantial damage yet, in the case of Lewisite exposure, recover relatively quickly. However, Lewisite is not an alkylating agent. Sulphur mustard, in contrast, generates characteristically slow healing lesions, most probably because of its ability to alkylate cell types that normally would be involved in skin regenerative processes. PMID- 14745846 TI - Protective effects of caffeic acid phenethyl ester on doxorubicin-induced cardiotoxicity in rats. AB - The prevention of doxorubicin (DXR)-induced cardiotoxicity may be helpful to improve future DXR therapy. The aim of this study was to investigate the cardio protective effects of caffeic acid phenethyl ester (CAPE), an antioxidant agent, on DXR-induced cardiotoxicity. Rats were divided into three groups and treated with saline, DXR and DXR + CAPE. Rats were treated with CAPE (10 micromol x kg( 1) day(-1) i.p.) or saline starting 2 days before a single dose of DXR (20 mg x kg(-1) i.p.). Ten days later, haemodynamic measurements were performed and the hearts were excised for biochemical analyses and microscopic examination. The heart rate and mean blood pressure were higher and the pulse pressure was lower in the DXR group than in the other two groups. The administration of DXR alone resulted in higher myeloperoxidase activity, lipid peroxidation and protein carbonyl content than in the other groups. The activities of superoxide dismutase and catalase were higher in DXR and DXR + CAPE groups than in the saline group. Rats in the DXR + CAPE group had increased catalase activity in comparison with the DXR group and high glutathione peroxidase activity in comparison with the other two groups. There was severe disruption of mitochondrial fi ne structure in the electron microscopy of the DXR group. In contrast, myocardial microscopy appeared nearly normal in the DXR + CAPE group (as de fi ned at the electron microscopic level). In light of these in vivo haemodynamic, enzymatic and morphological results, we conclude that CAPE pretreatment significantly attenuated DXR-induced cardiac injury, possibly with its antioxidant effects. PMID- 14745847 TI - Adrenal toxicity in dogs and cats as a contributing cause of hormonal and immune destabilization. AB - The adrenal cortex is regarded as the organ most vulnerable to toxicity within the endocrine system. The production of cortisol, among the many steroidal hormones produced by the cortex, may suffer as a result. In a veterinary clinical practice, household dogs and cats with a wide variety of diseases ranging from allergies to cancer commonly have a cortisol deficiency or defect that triggers endocrine imbalances and immune system destabilization. The causes of deficient cortisol are linked primarily to genetics but also to acquired adrenal damage likely stemming from environmental toxins. An innovative blood test to determine relevant endocrine-immune imbalances in pets and a treatment method based on low dosage steroidal medication, as a form of cortisol replacement therapy, are described. Despite a prevailing reluctance to use steroidal medications long term because of the fear of side effects, extended and even life-time usage of these medications at low, physiologic dosages has been applied successfully for decades and appears to be gaining wider acceptance. The validity of a combined testing and treatment method for humans based on the veterinary model deserves investigation as a tool with which to identify and correct toxic damage to adrenal function. PMID- 14745848 TI - Low-level exposure of guinea pigs and marmosets to sarin vapour in air: lowest observable-adverse-effect level (LOAEL) for miosis. AB - The purpose of this pilot study was to indicate, for low-level exposure of conscious guinea pigs and marmoset monkeys to sarin vapour in air, the lowest observable-adverse-effect level (LOAEL) of sarin for miosis. This is the concentration x time (C.t) value (t = 5 h) of exposure at which miosis becomes significant. The ratio of pupil and iris diameters, measured on digital photographs taken on-line during exposure, was calculated as a measure for miosis. The exposure concentrations were in the range 7-150 microg x m(-3) and the exposure times needed to achieve significant miosis were in the range 10-300 min. Both vehicle- and pyridostigmine-pretreated animals were used in the experiments. The latter pretreatment resulted in ca. 30% inhibition of erythrocyte acetylcholinesterase in both species. In vehicle-pretreated guinea pigs and marmosets the pupil size was decreased significantly (P < 0.05) at sarin doses of 1.8 +/- 0.3 and 2.5 +/- 0.8 mg x min x m(-3), respectively. In pyridostigmine-pretreated guinea pigs and marmosets the pupil size was affected significantly (P < 0.05) at 1.8 +/- 0.5 and 3.0 +/- 0.8 mg x min x m(-3), respectively. Evidently there is no significant influence of pyridostigmine pretreatment on the LOAEL. These data were addressed in light of the recommended occupational and detection limits for sarin vapour in air. It was concluded that miosis will occur during low-level sarin exposure at levels that are not detectable by the currently fielded alarm systems, assuming that humans are as sensitive for sarin vapour in air as guinea pigs and marmosets. PMID- 14745849 TI - Mouse strain differences in eosinophilic airway inflammation caused by intratracheal instillation of mite allergen and diesel exhaust particles. AB - Response differences by different strains of mice towards house dust mites (Dermatophagoides farinae) or diesel exhaust particles (DEP) were investigated. Mouse strains BALB/c, ICR and C3H/He received 1 micro g of D. farinae or 1 microg of D. farinae + 50 microg of DEP intratracheally four times at 2-week intervals. Dermatophagoides farinae treatment caused the recruitment of eosinophils and lymphocytes. The order of magnitude of the eosinophilic airway inflammation was BALB/c < ICR < C3H/He mice. The protein levels of eotaxin and IL-5 in lung tissues correlated with the manifestations of eosinophilic airway inflammation by D. farinae administration. Diesel exhaust particles aggravated the manifestation of the eosinophilic inflammation through goblet cell proliferation in the airway and enhanced the local expression of eotaxin and IL-5 in all three strains of mice. The levels of eotaxin and IL-5 in lung tissues corresponded to the pathological changes caused by D. farinae + DEP. The increasing order of production levels of antigen-specific IgG1 by D. farinae or D. farinae + DEP was BALB/c < ICR < C3H/He mice. The significant adjuvant effect of DEP on IgG1 production was observed in the C3H/He mice (P < 0.05). These results suggest that the murine strain differences in the production of eosinophilic airway inflammation by D. farinae + DEP are related to differences in local expression of IL-5 and eotaxin. The enhancing effects of DEP may be mediated by a cytokine increase in the local expression. Antigen-specific IgG1 may be an important immunoglobulin in the pathogenesis of allergic asthma enhanced by DEP. PMID- 14745850 TI - Assessment of the skin sensitization potency of eugenol and its dimers using a non-radioisotopic modification of the local lymph node assay. AB - Allergic contact dermatitis is a serious health problem. There is a need to identify and characterize skin sensitization hazards, particularly with respect to relative potency, so that accurate risk assessments can be developed. For these purposes the murine local lymph node assay (LLNA) was developed. Here, we have investigated further a modi fi cation of this assay, non-radioisotopic LLNA, which in place of tritiated thymidine to measure lymph node cell proliferation employs incorporation of 5-bromo-2'-deoxyuridine. Using this method we have examined the skin sensitizing activity of eugenol, a known human contact allergen, and its dimers 2,2'-dihydroxyl-3,3'-dimethoxy-5,5'-diallyl-biphenyl (DHEA) and 4,5'-diallyl-2'-hydroxy-2,3'-dimethoxy phenyl ether (DHEB). Activity in the guinea pig maximization test (GPMT) also measured. On the basis of GPMT assays, eugenol was classified as a mild skin sensitizer, DHEA as a weak skin sensitizer and DHEB as an extreme skin sensitizer. In the non-radioisotopic LLNA all chemicals were found to give positive responses insofar as each was able to provoke a stimulation index (SI) of >or=3 at one or more test concentrations. The relative skin sensitizing potency of these chemicals was evaluated in the non radioisotopic LLNA by derivation of an ec(3) value (the concentration of chemical required to provoke an SI of 3). The ec(3) values calculated were 25.1% for eugenol, >30% for DHEA and 2.3% for DHEB. Collectively these data suggest that assessments of relative potency deriving from non-radioisotopic LLNA responses correlate well with evaluations based on GPMT results. These investigations provide support for the proposal that the non-radioisotopic LLNA may serve as an effective alternative to the GPMT where there is a need to avoid the use of radioisotopes. PMID- 14745851 TI - Reflecting in tranquility: service on the National Institute of Nursing Research Initial Review Group. PMID- 14745852 TI - Korean women's attitudes toward physical activity. AB - In this study attitudes toward physical activity of three groups of Korean women were explored using a feminist qualitative research design. Seventeen healthy Korean women, 11 Korean women at risk of muscular atrophy, and 16 Korean immigrant women were recruited using a purposive sampling method. In-depth interviews using an interview guide were audiotaped and transcribed. Data were analyzed using thematic analysis. The findings indicate that the women viewed physical activity holistically, that death was viewed as the opposite of physical activity, that exercise was differentiated from physical activity, that exercise was connected to health, and that the women rarely participated in exercise because of their busy lives. The findings confirm the importance of considering the psychosocial contexts of attitudes toward health behavior. PMID- 14745853 TI - Depression burden, psychological adjustment, and quality of life in women with breast cancer: patterns over time. AB - The purpose of this study was to examine how level of depression burden influences women's psychological adjustment and quality of life over time and how depression burden interacted with a community-based oncology support program to influence psychological adjustment and life quality. Participants were 169 women who completed a side effects checklist at three data collection points. Women were divided into two groups based on their depression burden scores: 123 women reporting no burden, and 46 women reporting high depression burden. For psychological adjustment, there were significant interaction effects for intervention by time and for intervention by depression burden by time and significant main effects for depression burden. For life quality, there was a significant interaction effect for intervention by time and a significant main effect for depression burden. The findings document the negative impact of depression burden on psychological adjustment and life quality. Oncology support interventions can be effective in reducing this negative impact. PMID- 14745854 TI - Resilience to risk-taking behaviors in impoverished African American girls: the role of mother-daughter connectedness. AB - Rates of risk behaviors (e.g., violence, substance use) for impoverished girls are exceedingly high. Some view their future pessimistically, decreasing their resilience to avoid risky behaviors. Others resist such behaviors. Connectedness with an adult promotes this resilience, but how is unclear. The purpose of this secondary analysis was to investigate the relationships among connectedness to mother, time perspective, and resilience to risk-taking behaviors in impoverished African American girls ages 11-15. Structural equation modeling was used to cross validate this model. In contrast to the results found in earlier studies, no direct relationship emerged between maternal connectedness and resilience. Instead, future time perspective was the key mediator between connectedness and resilience. These findings suggest that an important aspect of interventions to foster resilience to risk behaviors in young girls could be to assist their mothers in developing a connected relationship with their daughters that would promote a future time perspective. PMID- 14745855 TI - Alzheimer's disease caregiving information and skills. Part II: family caregiver issues and concerns. AB - In recent years researchers have carried out an increasing number of clinical trials with family caregivers of the elderly. The results of these interventions have suggested that caregiver skill-building interventions may be more effective than information/support interventions. Although researchers have given considerable attention to the information and support needed by family caregivers, less is known about how this information and support translates into caregiver skills or into changes in behavior. This is the second in a series of three articles on a study in which researchers used qualitative methods to analyze summaries from the group component of a larger caregiver clinical trial. In this article we describe caregiver issues and concerns in five major areas: (a). dealing with change, (b). managing competing responsibilities and stressors, (c). providing a broad spectrum of care, (d). finding and using resources, and (e). experiencing emotional and physical responses to care. We identify specific skills needed by caregivers in addressing these caregiving issues and concerns, and we compare and contrast skilled with less skilled caregivers. Further work is needed concerning the focus and methods of future caregiver skill-based interventions. PMID- 14745856 TI - The life closure scale: additional psychometric testing of a tool to measure psychological adaptation in death and dying. AB - The purpose of this study was to conduct additional psychometric testing on an instrument designed to measure psychological adaptation in end-of-life populations across a wide spectrum of terminal illnesses. A sample of 20 participants completed initial testing of the Life Closure Scale (LCS); however, its usefulness was limited by the small sample size. A larger sample of 113 home hospice individuals who met established criteria and who gave informed consent completed the 27-item LCS for additional psychometric testing. Cronbach's alphas and correlation coefficients were computed, and factor analysis was conducted to establish internal consistency reliability, theoretical clarity, and criterion related validity. The number of scale items was reduced to 20, with a total alpha of.87. Cronbach's alphas for the two subscales were.80 (self-reconciled) and.82 (self-restructuring). Item-total correlations for the subscales ranged from a low of.37 to a high of.68, with confirmatory factor analysis yielding two loadings. These findings lend credence to the usefulness of the LCS in measuring psychological adaptation in dying persons. PMID- 14745857 TI - Locating and retaining research participants for follow-up studies. AB - Two common pitfalls of longitudinal research are loss of participants over time and inability to locate participants whose contact information has changed. This article is based on our experiences in locating and retaining a sample of caregivers of persons with Parkinson's disease 8-10 years after we last contacted them. The strategies we used resulted in locating 86% of our sample and retaining 80% of those who were eligible. These strategies included asking participants for a backup contact, asking participants if they would be willing to be contacted again for a future study, making the most of existing search engines, keeping in touch, and being flexible, patient, and professional. PMID- 14745858 TI - A potential role for perioperative hepatic arterial infusion chemotherapy? PMID- 14745860 TI - Proliferative activity in primary breast carcinomas is a salient prognostic factor. AB - BACKGROUND: The goal of the current study was to investigate the prognostic impact of proliferative activity, together with the other classic clinicopathologic prognostic factors (tumor size, tumor grade, receptor status, ploidy, and lymph node status), in breast carcinoma by counting mitoses and evaluating S phase fraction (SPF) in fresh and frozen tumor samples. METHODS: From March 1, 1990, to July 1, 1999, a total of 1984 previously untreated invasive breast carcinoma samples were snap-frozen for flow cytometry. RESULTS: After multivariate analysis incorporating all classic prognostic factors, SPF combined with mitotic activity (i.e., proliferative activity) remained the sole prognostic factor in the lymph node-negative group; proliferative activity was accompanied by tumor size as a prognostic factor in patients with lymph node positive disease and by lymph node status, lymphatic invasion, and receptor status in the overall population. The predictive value of proliferative activity was superior to that of the reference standards (classic prognostic predictors according to the guidelines of our institution [common oncology practice] and the St. Gallen classification). A review of the literature, focusing on series in which fresh material was used, allowed us to demonstrate that there is widespread agreement regarding the correlation between SPF and prognosis, even after multivariate analysis. CONCLUSIONS: S phase fraction is a valuable predictor of survival and can confidently be assessed in approximately 80% of cases. In conjunction with mitotic activity, SPF should become a prognostic factor that is used in daily practice by oncologists for the management of breast carcinoma. PMID- 14745859 TI - New agents in acute myeloid leukemia and other myeloid disorders. AB - Over the past several decades, improvements in chemotherapeutic agents and supportive care have resulted in significant progress in treating patients with acute myeloid leukemia (AML). More recently, advances in understanding the biology of AML have resulted in the identification of new therapeutic targets. The success of all-trans-retinoic acid in acute promyelocytic leukemia and of imatinib mesylate in chronic myeloid leukemia have demonstrated that targeted therapy may be more effective and less toxic when well defined targets are available. At the same time, understanding mechanisms of drug resistance and means to overcome them has led to modification of some of the existing cytotoxic agents. Rational design and conduct of clinical trials is necessary to ensure that the full potential of these new agents is realized. PMID- 14745861 TI - Effects of pregnancy after treatment for breast carcinoma on survival and risk of recurrence. AB - BACKGROUND: The goal of the current study was to assess the effect of pregnancy on the subsequent risk of recurrence after treatment for breast carcinoma, adjusting for established prognostic factors. METHODS: Between 1974 and 1998, 383 patients age < or =35 years were treated for breast carcinoma with adjuvant chemotherapy at The University of Texas M. D. Anderson Cancer Center (Houston, TX). The median follow-up period was 13 years. Of these, 13 patients were excluded from analysis, as no history was available regarding pregnancy; 240 (65%) were >30 years old; 47 (13%) had at least 1 pregnancy after therapy; 32 had full-term pregnancies; 10 had spontaneous or elective abortions; 4 had miscarriages; and 1 had a premature delivery. Estrogen receptor (ER) status, lymph node involvement, and disease stage were evaluated as potential risk factors for recurrence. Information on ER status was unavailable for 123 (33%) patients. RESULTS: Patients who experienced a pregnancy tended to have earlier stage disease (Stage I/II: 80% vs. 73%), fewer positive lymph nodes (<4: 87% vs. 52%), more ER negativity (68% vs. 58%), and younger age (<30 years: 57% vs. 32%) than patients who did not. The incidence of disease recurrence was 23% for women who experienced a pregnancy and 54% for women who did not. The hazard ratio (using the multivariate Cox proportional hazards model) for disease recurrence in patients with posttreatment pregnancy was 0.71 (P=0.4). CONCLUSIONS: In the current study population, pregnancy was not associated with an increased risk of disease recurrence or poorer survival in patients previously treated for breast carcinoma. PMID- 14745862 TI - Noninflammatory breast carcinoma with skin involvement. AB - BACKGROUND: According to the TNM Supplement, only breast carcinoma with macroscopic 'classic' skin changes (e.g., edema, peau d'orange, ulceration) should be placed in the T4 category; the classification of tumors with histologically proven skin involvement but no clinical skin changes or only discreet changes should be based on the size of the tumor (T1-3). To the authors' knowledge, no data supporting these recommendations have been reported to date. METHODS: Seventy-six patients with noninflammatory breast carcinoma and histologically proven skin involvement were classified based on the degree of their skin involvement. Fifty patients (66%) presented with clinically obvious skin involvement (Group A), and 26 patients (34%) had breast carcinoma with histologically proven skin involvement but without clinical skin changes (Group B). Reclassification was undertaken to assess the malignant potential of tumors independent of the morphologic parameter 'skin involvement'. RESULTS: Patients in Group A presented significantly more frequently with extensive disease at regional or distant sites (Stages IIIC and IV; P=0.009). The clinical outcome of patients in Group B was significantly more favorable (P=0.0003). The adjusted 3 year survival rates in Group A and Group B were 46.7% and 92.3%, respectively, and the 5-year rates were 38.1% and 83.7%, respectively. Patients in Group B were found to have significantly better (P=0.036) distant recurrence-free survival (DRFS) rates. The DRFS rate at 3 years was 63.6% in Group A, compared with 91.7% in Group B, and the DRFS rate at 5 years was 56.9% in Group A, compared with 82.0% in Group B. Using a cutoff point of tumor size=3 cm, similar findings were made. CONCLUSIONS: Both study groups exhibited distinct clinical entities with significant differences in clinical course and prognosis. Cases with histologic skin involvement, relative to those with classic skin involvement, were found to have much less malignant potential. This corroborates the recommendation that cases with only histologic skin involvement should not be placed in the T4 category of the TNM classification. PMID- 14745863 TI - Breast magnetic resonance image screening and ductal lavage in women at high genetic risk for breast carcinoma. AB - BACKGROUND: Intensive screening is an alternative to prophylactic mastectomy in women at high risk for developing breast carcinoma. The current article reports preliminary results from a screening protocol using high-quality magnetic resonance imaging (MRI), ductal lavage (DL), clinical breast examination, and mammography to identify early malignancy and high-risk lesions in women at increased genetic risk of breast carcinoma. METHODS: Women with inherited BRCA1 or BRCA2 mutations or women with a >10% risk of developing breast carcinoma at 10 years, as estimated by the Claus model, were eligible. Patients were accrued from September 2001 to May 2003. Enrolled patients underwent biannual clinical breast examinations and annual mammography, breast MRI, and DL. RESULTS: Forty-one women underwent an initial screen. Fifteen of 41 enrolled women (36.6%) either had undergone previous bilateral oophorectomy and/or were on tamoxifen at the time of the initial screen. One patient who was a BRCA1 carrier had high-grade ductal carcinoma in situ (DCIS) that was screen detected by MRI but that was missed on mammography. High-risk lesions that were screen detected by MRI in three women included radial scars and atypical lobular hyperplasia. DL detected seven women with cellular atypia, including one woman who had a normal MRI and mammogram. CONCLUSIONS: Breast MRI identified high-grade DCIS and high-risk lesions that were missed by mammography. DL detected cytologic atypia in a high-risk cohort. A larger screening trial is needed to determine which subgroups of high-risk women will benefit and whether the identification of malignant and high-risk lesions at an early stage will impact breast carcinoma incidence and mortality. PMID- 14745864 TI - Potential applicability of balloon catheter-based accelerated partial breast irradiation after conservative surgery for breast carcinoma. AB - BACKGROUND: Balloon catheter-based accelerated partial breast irradiation (APBI) is an alternative to whole-breast external-beam irradiation during breast conserving therapy (BCT) for breast carcinoma, but it is limited by the size of the segmental mastectomy cavity. There are scant data on the average or optimal volume of resection (VR) in BCT. The objective of the current study was to evaluate the percentage of patients who would be eligible for balloon catheter based APBI based on the selection criteria of the American Society of Breast Surgeons and the surgical VR. METHODS: The authors reviewed the medical records of 443 patients with ductal carcinoma in situ (DCIS) or invasive carcinoma treated with BCT. Patient treatment and pathologic data were analyzed to assess VR and eligibility for APBI. RESULTS: BCT was performed for 178 patients with DCIS and 267 patients with invasive breast carcinoma. The majority of invasive carcinomas (63.3%) were infiltrating ductal carcinomas. The median overall lumpectomy volume was 67.61 cm3, with no significant difference between DCIS and invasive carcinoma (P>0.05). Although the majority (62.9-82.0%) of patients met the individual selection criteria for APBI, only 27.4% of the cohort was found to be eligible for any type of APBI when the selection criteria were considered together. Based on VR, only approximately one-half of the patients initially eligible for APBI would be candidates for immediate balloon catheter-based APBI using the 70 cm3 balloon device (13.3%). However, with the new, larger 125 cm3 balloon device, approximately three-fourths of patients initially eligible for APBI would be eligible for balloon catheter-based APBI at the time of the initial surgical procedure (20.7%). Although not evaluated in the current study, shrinkage of the lumpectomy cavity with time may increase the number of patients eligible based strictly on VR criteria. Patients with a very large VR (> or =125 cm3) were more likely to have invasive carcinoma (P=0.02; hazard ratio [HR], 7.4) and tumors > or =5 cm on final pathology (P<0.01; HR, 22.0). CONCLUSIONS: Approximately one-fifth to one-fourth of patients presenting for BCT may be eligible for balloon catheter-based APBI according to accepted national guidelines and VR. VR must be considered when selecting patients for balloon catheter-based APBI, because a minority of patients will have a lumpectomy cavity that exceeds the size limit of the current balloon device. PMID- 14745865 TI - Prognostic significance of phosphorylated P38 mitogen-activated protein kinase and HER-2 expression in lymph node-positive breast carcinoma. AB - BACKGROUND: Chemotherapy-induced p38 mitogen-activated protein kinase (MAPK) phosphorylation reportedly leads to increased apoptosis in breast carcinoma cells. The goals of the current study were to assess the incidence of activated phosphorylated p38 MAPK (P-p38) expression in invasive breast carcinoma, correlate expression of P-p38 MAPK with HER-2, and estimate the prognostic value of this marker in patients with lymph node-positive breast carcinoma treated with adjuvant chemotherapy. METHODS: P-p38, HER-2, and Ki-67 were measured using immunohistochemistry (peroxidase method) in 96 patients with lymph node-positive breast carcinoma treated with adjuvant fluorouracil, doxorubicin, and cyclophosphamide chemotherapy. All markers were measured in the primary tumors, before the initiation of adjuvant chemotherapy. The median follow-up period was 11 years after initial cancer surgery. P-p38 MAPK expression was scored visually and quantified using an image analyzer. RESULTS: The rate of P-p38 MAPK expression ranged from 19-24%, depending on the scoring system used. There was a trend toward shorter progression-free survival (PFS) for patients whose tumors expressed high levels of P-p38 MAPK, although the difference was not statistically significant (P=0.39). PFS was shorter in patients whose tumors overexpressed P-p38 MAPK and had a high level of Ki-67 (P=0.04). In HER-2 negative patients, P-p38 MAPK overexpression was associated with a shorter PFS (P=0.05). CONCLUSIONS: P38 MAPK phosphorylation occurred in 20% of primary breast carcinomas and may be associated with poor outcome in patients with lymph node positive breast carcinoma. Further studies are needed to define the interaction between P-p38 MAPK and HER-2 expression in breast carcinoma. PMID- 14745866 TI - A retrospective analysis of health care costs for bone fractures in women with early-stage breast carcinoma. AB - BACKGROUND: In this retrospective data base study, the authors sought to estimate direct costs for bone fractures in women age > or =65 years with early-stage breast carcinoma and to compare those costs with treatment costs for bone fractures in older women without early-stage breast carcinoma. METHODS: Direct costs for bone fractures in patients with early-stage breast carcinoma, which consist of excess treatment costs for bone fracture and excess costs of long-term care for bone fracture, were evaluated by using the 1997-1998 Standard Analytical File. The statistical significance of the difference in inpatient costs, medical treatment costs, and long-term care admission rates were determined with the t test and the Fisher chi-square test, respectively. RESULTS: For older women with early-stage breast carcinoma, the direct costs for bone fracture were estimated at $45,579, and 57% of those costs came from treating the bone fracture (32% came from inpatient hospital costs, and 25% came from noninpatient hospital costs), 25% came from other excess treatment costs, and 18% came from excess long-term care costs. The women who had early-stage breast carcinoma and sustained bone fracture did not differ significantly from the women without early-stage breast carcinoma who sustained a bone fracture. CONCLUSIONS: Bone fracture was associated with high direct costs in older women with early-stage breast carcinoma. Additional research should include appropriate, incidence-based studies to investigate the potential benefit of an intervention for preventing bone fracture in this increasingly large patient population. PMID- 14745867 TI - Should decisions on internal mammary lymph node irradiation be based on current lymphoscintigraphy techniques for sentinel lymph node identification? AB - BACKGROUND: Treatment of internal mammary lymph node (IMN) metastases remains controversial because of the difficulty in predicting involvement, potential treatment-related morbidity, and questionable efficacy. Lymphoscintigraphy with sentinel lymph node biopsy offers a means to identify occult involvement of IMN, allowing appropriate patient selection for IMN treatment. METHODS: The authors retrospectively reviewed 262 lymphoscintigraphies (LS) of 248 patients treated at the University of Florida (Gainesville, FL) between 1998 and 2002. Tumor characteristics were assessed for their value in predicting IMN drainage and their association with IMN radiation. RESULTS: Lymph flow to the IMN was documented with LS in 23 of 262 tumor specimens (9%). Flow to the IMN was not correlated with any of the five factors: tumor location, tumor size, lymphovascular invasion, pathologic lymph node status, and laterality of the involved breast (right vs. left breast). Identification of IMN flow increased from 5.7% to 10.1% with the use of a deep injection technique. IMN radiotherapy was used more frequently in patients with larger tumors (15 of 188 in Tis/T1 vs. 31 of 70 in T2-T4; P<0.0001) and positive lymph nodes (17 of 91 in lymph node negative patients vs. 28 of 66 in lymph node-positive patients; P<0.0001). In patients with T2N0 tumors (n=32), IMN radiotherapy was used more frequently with medial tumors (5 of 11 [45%]) than with lateral tumors (4 of 21 [19%]). CONCLUSIONS: The incidence of flow to the IMN documented with the current LS technique was low compared with other LS and extended radical mastectomy series. Histopathologic information was obtained for the sentinel IMN when IMN flow was identified on the LS. In the absence of histopathologic information, treatment decisions should continue to be based on clinical factors known to be correlated with occult IMN involvement. PMID- 14745868 TI - Association of ampullary and colorectal malignancies. AB - BACKGROUND: Because of the similarities in terms of carcinogenesis and natural history between cancer of the ampulla of Vater and colorectal cancer, the authors examined whether ampullary and colorectal malignancies occur in the same individuals at increased rates. METHODS: We used data from the Surveillance, Epidemiology, and End Results (SEER) Program of the National Cancer Institute for the period from January 1973 through December 1999. Person-years of follow-up for patients with ampullary (or colorectal) cancer were used to calculate the expected number of cases of colorectal (or ampullary) cancer as a second primary malignancy. Standardized incidence ratios (SIRs) with 95% confidence intervals (CIs) were calculated using Byar limits and assuming a Poisson distribution. RESULTS: The authors identified 2043 white patients with ampullary cancer who were included in the SEER registry between 1973 and 1999. Over an aggregate 5674 person-years of follow-up, 30 patients, compared with an expected 14, developed colorectal cancer, yielding an overall SIR of 2.14 (95% CI, 1.45-3.06). Similarly, 57 of 262,066 white patients with colorectal cancer developed ampullary cancer over an aggregate 1,270,255 person-years of follow-up, yielding an SIR of 2.18 (95% CI, 1.69-2.85). CONCLUSIONS: Patients with ampullary cancer are at increased risk for a second primary colorectal malignancy, and patients with colorectal cancer are at increased risk for a second primary ampullary malignancy. These findings suggest that ampullary and colorectal malignancies share common environmental and/or genetic risk factors. PMID- 14745869 TI - Treatment of advanced colorectal carcinoma with oxaliplatin and capecitabine: a phase II trial. AB - BACKGROUND: The current study was designed to evaluate the antitumor activity and toxicity of capecitabine and oxaliplatin in previously untreated patients with advanced colorectal carcinoma. The primary endpoint of the study was to determine the objective response rate, and a secondary endpoint was to measure the time to disease progression. METHODS: A 2-stage trial was planned with an accrual goal of 35 patients. The treatment included oxaliplatin given at a dose of 130 mg/m2 on Day 1 of each 3-week cycle. Initially, capecitabine at a dose of 2000 mg/m2/day in 2 divided doses was given on Days 1-14 of each cycle, but this was reduced to a dose of 1500 mg/m2/day because of toxicity. Patients were followed by computed tomography scans every two cycles to evaluate treatment response, and toxicity was monitored. RESULTS: The first 13 patients on the trial received the higher dose of capecitabine. Although 5 responses (38.5%) were noted, 5 patients were hospitalized with diarrhea and dehydration. This toxicity led to a decrease in the dose of capecitabine to 1500 mg/m2/day and an additional 35 patients were treated. At the lower dose, the partial response rate was 37.1% (95% confidence interval [95% CI], 21.5-55.1%). The estimated median progression-free survival was 6.9 months (95% CI, 4.4-8.2 months). At the lower dose, four patients were hospitalized with diarrhea/dehydration (with one death reported), one with febrile neutropenia, and one with ventricular fibrillation. Overall, Grade (according to version 2.0 of the National Cancer Institute Common Toxicity Criteria) 3-4 diarrhea was reported to develop in 20% of those patients treated at the capecitabine dose of 1500 mg/m2/day compared with 62% of patients treated at the dose of 2000 mg/m2/day. CONCLUSIONS: The combination of oxaliplatin and capecitabine is an active and convenient regimen for the treatment of patients with advanced colorectal carcinoma and should be compared with other front-line regimens as therapy for disease. PMID- 14745870 TI - The relationship of increasing radiotherapy dose to reduced distant metastases and mortality in men with prostate cancer. AB - BACKGROUND: The association of increasing radiotherapy (RT) dose with reduced biochemical failure (BF) is accepted widely. However, there is little direct evidence that dose escalation has an impact on distant metastasis (DM) or overall mortality (OM). These associations were examined in the current study. METHODS: The outcome of 835 patients who were treated at the Fox Chase Cancer Center (Philadelphia, PA) between 1989 and 1997 using 3-dimensional, conformal RT alone (median dose, 74 Gray [Gy]) was analyzed. Stepwise multivariate Cox proportional hazards regression analyses (MVAs) were performed with RT dose included as a covariate along with log-transformed initial pretreatment PSA level, Gleason score, palpation T status, age, and year of treatment (YOT), where indicated. To minimize the effect of YOT, an analysis was performed on a subgroup of 363 patients who were treated prior to 1994. RESULTS: With a median follow-up of 64 months, there were 220 PSA failures, 44 distant metastases, and 162 deaths. In MVA, RT dose (as a continuous variable) was a significant predictor for BF, DM, and OM. When YOT was included as a covariate, it was related strongly to all endpoints, and the correlations of RT dose with DM and OM were lost. When the effect of YOT was minimized by limiting the MVA to patients who were treated prior to 1994, RT dose again emerged as a significant predictor of DM. CONCLUSIONS: Escalation of RT dose reduced the rates of BF, DM, and OM significantly in patients with prostate cancer. The inclusion of YOT had a pronounced effect on these correlations that may confound interpretation. PMID- 14745871 TI - Restaging of recurrent cervical carcinoma with dual-phase [18F]fluoro-2-deoxy-D glucose positron emission tomography. AB - BACKGROUND: The clinical value of positron emission tomography (PET) with [18F]fluoro-2-deoxy-D-glucose (FDG) for primary staging in cervical carcinoma appears to be promising. The authors sought to evaluate the diagnostic efficacy and benefit of PET in restaging cervical carcinoma at the time of first recurrence. METHODS: Forty patients with cervical carcinoma who experienced confirmed treatment failure but who were feasible candidates for curative salvage therapy were enrolled prospectively in the current study. Restaging was performed with PET and with computed tomography and/or magnetic resonance imaging (CT/MRI). Dual-phase PET was performed by adding 3-hour-delayed images to the 40-minute scans. The results of the PET and CT/MRI scans were compared. Lesion status was determined by pathologic findings or by clinical follow-up. The receiver operating characteristic curve method with calculation of area under the curve (AUC) was used to evaluate diagnostic efficacy. The primary endpoint was percent improvement in restaging (with improvement indicated by treatment modification) after PET. The secondary endpoint was 2-year overall survival among study participants compared with comparable previously treated patients who did not undergo disease restaging with PET. RESULTS: Twenty-two patients (55%) had their treatment modified due to PET findings. PET was significantly superior to CT/MRI (sensitivity: 92% vs. 60%; AUC: 0.962 vs. 0.771; P<0.0001) in identifying metastatic lesions. For individuals receiving primary surgery, a significantly better 2-year overall survival rate was observed among study participants compared with patients who underwent disease restaging without PET (HR, 0.21 [95% confidence interval, 0.05-0.83]; P=0.020). CONCLUSIONS: Dual-phase FDG-PET is superior to CT/MRI in the restaging of recurrent cervical carcinoma. Restaging with PET provides benefit by allowing the physician to offer optimal management of recurrent cervical carcinoma. PMID- 14745872 TI - Prognostic significance of intratumoral lymphangiogenesis in squamous cell carcinoma of the oral cavity. AB - BACKGROUND: Clinicopathologic data demonstrated that the lymphatic system is the main route for solid tumor metastasis. However, the effect of intratumoral lymphangiogenesis (IL) on prognosis in oral carcinoma is still unknown because, until recently, no reliable markers for lymphatic endothelium were available. The current study analyzed the lymphatic vessels in tumor tissue specimens of patients with primary oral carcinoma using the new marker, PA2.26. METHODS: The authors investigated IL in surgical tissue samples of 61 patients with early stage (Stages I-II) oral carcinoma. The tissue specimens were stained for PA2.26 and the correlation between IL and relevant parameters was analyzed by the Pearson chi-square test. In a univariate analysis using the Kaplan-Meier method, IL was analyzed against survival and disease-free period. Statistical significance of differences between distributions was studied by the log-rank test. Clinicopathologic parameters, including IL, were entered in a multivariate analysis to determine independent prognostic significance. RESULTS: Thirty-three patients had IL. In the follow-up, a strong association was found between IL and locoregional recurrence (30.3 % of the patients with IL and 7.1% of the patients without IL). The presence of IL did not correlate significantly with the pT classification, primary location, or tumor differentiation. IL was found to have no influence on overall survival in univariate analysis, but there was significant association between IL and disease-free survival (P=0.03). Multivariate analysis revealed IL to be the sole independent factor influencing disease-free interval (P=0.02). CONCLUSIONS: These results suggested that IL is associated with locoregional disease recurrence in early-stage oral carcinoma. The presence of IL was a useful discriminator in predicting the outcome of patients with absence of lymph node metastasis. PMID- 14745873 TI - Gamma radiation sensitivity and risk of malignant and benign salivary gland tumors: a pilot case-control analysis. AB - BACKGROUND: The salivary gland is a highly radiosensitive organ. Exposure to gamma radiation is a risk factor for both malignant (MSTs) and benign salivary gland tumors (BSTs), but the exact mechanisms remain unknown. The objectives of the current study were to determine whether gamma radiation-induced chromatid breaks increase the risk of MSTs and BSTs and whether there is any difference in risk between these two diseases. METHODS: The authors performed a pilot case control study of 57 patients with salivary gland diseases (45 patients with MSTs and 12 patients with BSTs) and 105 cancer-free controls. Peripheral blood lymphocytes from these participants were cultured and exposed to gamma radiation (1.5 grays). Five hours later, metaphase spread slides were evaluated. The chromatid breaks in 50 well-spread metaphase slides were counted to determine the average number of chromatid breaks per cell (b/c). RESULTS: Multivariate logistic regression analyses revealed that gamma radiation-induced b/c values greater than the median of the controls were a significant risk factor for salivary gland tumors (adjusted odds ratio [OR], 17.25; 95% confidence interval [CI], 4.92 60.49). The risk remained significant for MSTs (adjusted OR, 40.45; 95% CI, 5.27 310.17) but was of borderline significance for BSTs (adjusted OR, 4.73; 95% CI, 0.94-23.87) when these tumors were analyzed separately. CONCLUSIONS: In the current study, high levels of chromatid breaks in lymphocytes induced by gamma irradiation were associated with an independent risk for MSTs and were likely to increase the risk of BSTs. However, larger studies are needed to verify these findings. PMID- 14745874 TI - Intravenous itraconazole for prophylaxis of systemic fungal infections in patients with acute myelogenous leukemia and high-risk myelodysplastic syndrome undergoing induction chemotherapy. AB - BACKGROUND: Systemic fungal infections remain the leading cause of mortality in patients with newly diagnosed acute myelogenous leukemia (AML) and high-risk myelodysplastic syndrome (MDS). The objective of the current study was to determine whether intravenous itraconazole (i.v. ITRA) reduced the incidence of probable/proven fungal infections in this group of patients, and compare the results with those of a historic control group treated with fluconazole plus itraconazole capsules (F+I). METHODS: Patients with AML and high-risk MDS who underwent induction chemotherapy received 200 mg of i.v. itraconazole over 60 minutes every 12 hours during the first 2 days followed by 200 mg given i.v. once daily. RESULTS: One hundred patients were enrolled, 96 of whom were evaluable. Approximately 48% of the patients in the group of patients treated with i.v. ITRA as well as in the F+I group completed prophylaxis. Nine patients (9%) in the study group developed either proven/probable fungal infections (Candida glabrata in 5 patients, C. tropicalis in 1 patient, C krusei in 1 patient, and Fusarium in 2 patients) compared with 3 patients (4%) with proven fungal infection in the historic control group (C. tropicalis in 1 patient and Aspergillus in 2 patients). There were no significant differences noted between the two groups with regard to the percentage of patients who developed proven/probable or possible fungal infection as well as with regard to survival. These results also were obtained after adjusting for relevant prognostic factors (creatinine and bilirubin). The most common toxicity encountered with the use of i.v. ITRA was NCI Grade 3-4 hyperbilirubinemia (6%). CONCLUSIONS: Despite its theoretic advantages, the authors found no evidence that i.v. ITRA is superior to itraconazole capsules, at least when the latter is combined with fluconazole. PMID- 14745875 TI - Activity of interferon-alpha and isotretinoin in patients with advanced, refractory lymphoid malignancies. AB - BACKGROUND: Interferon-alpha (IFN-alpha) and retinoids have shown nonoverlapping toxicity and each has shown antitumor activity in patients with lymphoma. The aim of the current study was to assess the toxicity, safety, and efficacy of IFN alpha combined with isotretinoin in patients with advanced, refractory lymphoid malignancies. METHODS: Adults with biopsy-proven advanced lymphoid malignancy were treated. Patients with compromised bone marrow function (platelet counts as low as 30 x 10(9)/L) were eligible. Treatment was comprised of IFN-alpha at a starting daily dose of 3 mega units subcutaneously and isotretinoin orally starting at a dose of 1 mg/kg daily in 2 divided doses. RESULTS: Forty-four patients were evaluable. Their median age was 57 years (range, 18-82 years). Eighteen patients had advanced cutaneous T-cell lymphoma, 6 patients had peripheral T-cell lymphoma, 14 patients had Hodgkin disease, and 6 patients had a variety of other lymphoid malignancies. Patients with Hodgkin disease had received a median of 6 previous therapies (range, 3-12 therapies) and patients with other lymphoid malignancies had received a median of 4 previous therapies (range, 1-9 therapies). The median duration of treatment was 4 months (range, 0.25-38 months). The overall response rate was 38.6% (complete response in 5 patients [11.3%] and partial response in 12 patients [27.3%]). The median response duration was 3 months (range, 1-95+ months). The most common toxicities were low-grade fever, flu-like symptoms, and fatigue (IFN-alpha effects); dry mouth and skin and hypertriglyceridemia (cis-retinoic acid effects); and thrombocytopenia (which generally occurred in patients with low baseline platelet counts). CONCLUSIONS: IFN-alpha and isotretinoin combination therapy had antitumor activity and was well tolerated in heavily pretreated patients with lymphoid malignancies. PMID- 14745876 TI - Amphotericin B lipid complex as prophylaxis of invasive fungal infections in patients with acute myelogenous leukemia and myelodysplastic syndrome undergoing induction chemotherapy. AB - BACKGROUND: The optimal antifungal prophylactic regimen for patients with acute myelogenous leukemia (AML) or high-risk myelodysplastic syndrome (MDS) undergoing induction chemotherapy has yet to be identified. A prospective historical control study evaluated the efficacy and safety of amphotericin B lipid complex (ABLC) in this patient population. METHODS: Newly diagnosed patients with AML or high-risk MDS who were undergoing induction chemotherapy received prophylactic ABLC 2.5 mg/kg intravenously 3 times weekly. This treatment group was compared with a historical control group that had similar baseline characteristics and received prophylactic liposomal amphotericin B (L-AmB) 3 mg/kg 3 times weekly. The primary endpoint was the incidence of documented or suspected fungal infections during and up to 4 weeks after cessation of prophylaxis. Reported adverse events were used to assess tolerability. RESULTS: The overall efficacy of antifungal prophylaxis was similar in patients who received ABLC and patients who received L AmB (P=0.95). Among 131 ABLC-treated patients and 70 L-AmB-treated patients who were assessed for efficacy and safety, 49% of patients in each group completed therapy without developing a documented or suspected fungal infection. Documented fungal infections occurred in 5% of ABLC-treated patients and in 4% of L-AmB treated patients. Alternative antifungal strategies were required because of persistent fever or pneumonia of unknown pathogen in 28% and 32% of ABLC-treated and L-AmB-treated patients, respectively. Grade 3 and 4 adverse events, therapy discontinuations due to adverse events, and survival rates also were similar between treatment groups. CONCLUSIONS: ABLC and L-AmB appeared to have similar efficacy and were tolerated well as antifungal prophylaxis in patients with AML and high-risk MDS who were undergoing induction chemotherapy. PMID- 14745877 TI - Prophylactic hepatic arterial infusion chemotherapy for the prevention of liver metastasis in patients with colon carcinoma: a randomized control trial. AB - BACKGROUND: The liver is the most frequent site of recurrence after curative resection in patients with colon carcinoma. For liver metastasis, a high response rate can be achieved with hepatic arterial infusion (HAI) chemotherapy. In the current study, the authors administered 5-fluorouracil (5-FU) as adjuvant chemotherapy by HAI to patients with colon carcinoma without liver metastases and studied its effects on recurrence in the liver and survival. METHODS: A total of 316 patients with preoperative Stage II or Stage III colon carcinoma (according to the 1997 revision of the International Union Against Cancer TNM staging system) were randomly assigned to receive surgery plus 3-week continuous HAI of 5 FU or surgery alone. There were 305 eligible patients, of whom the 119 patients assigned to the HAI arm actually received 5-FU. The primary endpoint was disease free survival, whereas the secondary endpoints were overall survival and liver metastasis-free survival. Analysis was by intent to treat. RESULTS: There were no significant differences noted in morbidity between the two treatment arms. During the follow-up period (median, 59.0 months), the incidence of liver metastasis was significantly decreased in the HAI arm whereas there were no significant differences reported between the 2 arms with regard to the frequency of metastasis at other sites. In the HAI arm, the risk ratio for recurrence was 0.40 (95% confidence interval [95% CI], 0.24-0.64; P=0.0002), the risk ratio for death was 0.37 (95% CI, 0.21-0.67; P=0.0009), and the risk ratio for liver metastasis was 0.38 (95% CI, 0.22-0.66; P=0.0005). These differences were found to be significant only for patients with Stage III disease. Toxicities were mild. CONCLUSIONS: A schedule of 3-week HAI of 5-FU given as adjuvant chemotherapy to patients with Stage III colon carcinoma appeared to contribute to a significant decrease in the frequency of liver metastases and was associated with an improved survival rate. PMID- 14745878 TI - Desmoplastic and neurotropic melanoma. AB - BACKGROUND: Desmoplastic and neurotropic melanoma (DNMM) occasionally metastasizes to regional lymph nodes and extranodal sites. The value of sentinel lymph node biopsy (SLNB) has not been demonstrated clearly for patients with DNMM. The authors report on the utility of SLNB in the management of patients with DNMM. METHODS: The authors identified 33 patients with DNMM who were seen during a 5-year period in their institution who underwent lymphatic mapping and SLNB. Clinical and histopathologic data were reviewed. RESULTS: Thirty-three patients with DNMM underwent SLNB (mean Breslow depth, 4.0 mm; median, 2.8 mm). There were 25 male patients and 8 female patients with a median age of 61 years (range, 31-86 years). Fifty-two percent of tumors presented in the head and neck region, and 24% were associated with lentigo maligna. Four of 33 patients (12%) without clinical evidence of metastatic disease who underwent SLNB had at least 1 positive sentinel lymph node. No additional positive lymph nodes were found in subsequent therapeutic regional lymphadenectomy in any of these four patients. CONCLUSIONS: SLNB detected subclinical metastases of DNMM to regional lymph nodes. SLNB at the time of resection can provide useful information to guide early treatment and, coupled with lymphadenectomy in positive patients, may limit tumor spread and prevent recurrence at the draining lymph node basin. PMID- 14745879 TI - Temozolomide in the treatment of recurrent malignant glioma. AB - BACKGROUND: Options for chemotherapy at the time of recurrence in patients with malignant glioma are limited. The authors describe the efficacy and safety results of their institution's open-label, compassionate-use protocol of temozolomide for patients with recurrent malignant glioma. METHODS: Patients with recurrent malignant glioma at any time during recurrence were treated with oral temozolomide at a dose of 150 mg/m2 per day on a 5-day schedule every 28 days. If this dose was tolerated, then escalation to 200 mg/m2 was allowed. Clinical evaluations and assessments of tumor response were performed every 2 months. All patients or their surrogates signed approved Institutional Review Board consent forms. RESULTS: Among 213 patients who were treated, 33% had Grade 3 tumors, and 67% had Grade 4 tumors. The overall objective response rate was 16% in both of these patient groups; and an additional 51% and 30% of patients with Grade 3 and Grade 4 tumors, respectively, had stable disease as their best response. The 6 month progression-free survival rates were 41% and 18% for patients with Grade 3 and Grade 4 tumors, respectively. The median survival was 49 weeks for patients with Grade 3 tumors and 32 weeks for patients with Grade 4 tumors. The major toxicity was hematologic toxicity. In multivariate analysis, the Karnofsky performance score was a significant predictor of survival for patients with Grade 4 tumors. CONCLUSIONS: Temozolomide was well tolerated in patients with recurrent malignant glioma and had modest efficacy, even at the time of multiple recurrences. PMID- 14745880 TI - High-dose tamoxifen and sulindac as first-line treatment for desmoid tumors. AB - BACKGROUND: Desmoid tumors are mesenchymal nonmetastasizing neoplasms. Although rare in the general population, they are a common extracolonic manifestation of familial adenomatous polyposis (FAP). Because of high tumor recurrence rates, surgery has been less than satisfactory in the treatment of desmoid tumors. In the current study, high doses of tamoxifen in combination with sulindac were used to treat severe desmoid tumors to avoid surgery. METHODS: Since 1992, 25 patients at Heinrich Heine University (Dusseldorf, Germany) were treated with a combination of tamoxifen and sulindac. In the current study, 17 patients with FAP associated and 8 patients with sporadic desmoid tumors received 120 mg of tamoxifen and 300 mg of sulindac daily. Every 6 months, the protracted course of desmoid growth was measured by computed tomography and/or magnetic resonance imaging scans. Tumor responses were characterized as progressive disease, stable disease (SD), partial regression (PR), and complete regression (CR). RESULTS: Of the group of patients who received tamoxifen and sulindac as a primary treatment, all three patients with sporadic desmoid tumors demonstrated cessation of growth, and 10 of the 13 patients with FAP-associated tumors achieved either a PR or CR. In the sporadic desmoid tumor group, eight of nine patients developed tumor recurrences after undergoing surgery at other institutions. Of these, two patients had SD and two patients had a PR to CR. CONCLUSIONS: The patients with desmoid tumors who were managed conservatively with high-dose tamoxifen and sulindac had the best outcome. Desmoid tumor recurrence after surgery was high and in the FAP-associated tumor group, therapy with tamoxifen and sulindac was found to be less successful. Based on this experience, the authors recommended high-dose tamoxifen and sulindac as the primary treatment for patients with FAP associated desmoid tumors. However, to our knowledge, the best approach after surgical intervention for patients with sporadic desmoid tumors remains to be determined. PMID- 14745881 TI - Survival experience of black patients and white patients with bladder carcinoma. AB - BACKGROUND: Blacks are less likely than whites to develop bladder carcinoma. However, once they are diagnosed, black patients experience poorer survival. The authors investigated which factors were related to survival differences in black patients and white patients with bladder carcinoma stratified by extent of disease. METHODS: A population-based cohort of black patients with bladder carcinoma and a random sample of frequency-matched white patients with bladder carcinoma, stratified by age and gender, were identified through cancer registry systems in Atlanta, New Orleans, and San Francisco/Oakland. Patients had no previous cancer history and were ages 20-79 years at the time they were diagnosed with bladder carcinoma in 1985-1987. Medical records were reviewed at initial diagnosis, and 77% of patients were interviewed. Tumor grade, T classification, and other variables, including age, socioeconomic position, symptom duration, smoking history, and comorbidities, were recorded. Survival of black patients and white patients by extent of disease was modeled using Cox regression analysis. RESULTS: A greater proportion of black patients had histologic types of tumors that were associated with poorer survival. Among those with pure urothelial carcinoma, black patients had greater extent of disease at the time of diagnosis. Within specific extent-of-disease categories, there was some evidence of poorer survival for black patients with T2 tumors and strong evidence of poorer survival among those with T3 tumors compared with white patients. Black patients with muscle-invasive carcinoma who died within 6 months of diagnosis tended to present with life-threatening symptoms. Black patients and white patients did not differ with respect to diagnostic tests performed or therapy given. CONCLUSIONS: Black patients with bladder carcinoma had poorer survival due to greater extent of disease at diagnosis and a higher proportion of more aggressive histologies compared with white patients. Within urothelial carcinomas, by extent of disease (clinical/pathologic stage) these black/white survival differences were limited to patients with muscle invasion (T2 and T3 tumors). PMID- 14745882 TI - The cancer screening practices of adult survivors of childhood cancer: a report from the Childhood Cancer Survivor Study. AB - BACKGROUND: The current study characterized the self-reported cancer screening practices of adult survivors of childhood cancer. METHODS: A cohort of 9434 long term survivors of childhood cancer and a comparison group of 2667 siblings completed a 289-item survey that included items regarding cancer-screening practices. RESULTS: Overall, 27.3% of female respondents reported performing breast self-examination (BSE) regularly, 78.2% reported undergoing a Papanicolaou smear within the previous 3 years, 62.4% underwent a clinical breast examination (CBE) within the last year, and 20.9% had gotten a mammogram at least once in their lifetime. Approximately 17.4% of male respondents reported performing regular testicular self-examination (TSE). Women age > or =30 years who had been exposed to chest or mantle radiation therapy were more likely to report undergoing CBE (odds ratio [OR], 1.59; 95% confidence interval [95% CI], 1.32 1.92) and mammography (OR, 1.92; 95% CI, 1.47-2.56). Compared with the sibling comparison group, survivors demonstrated an increased likelihood of performing TSE (OR, 1.52; 95% CI, 1.22-1.85) or BSE (OR, 1.30; 95% CI, 1.10-1.52), of having undergone a CBE within the last year (OR, 1.18; 95% CI, 1.02-1.35), and of ever having undergone a mammogram (OR, 1.82; 95% CI, 1.52-2.17). CONCLUSIONS: The results of the current study demonstrate that the cancer screening practices among survivors of childhood cancer are below optimal levels. Primary care physicians who include childhood cancer survivors among their patients could benefit these individuals by informing them about future cancer risks and recommending appropriate evidence-based screening. PMID- 14745883 TI - Radiofrequency ablation: the experts weigh in. PMID- 14745884 TI - Fourteen-gauge needle core biopsy of mammographically evident radial scars: is excision necessary? PMID- 14745886 TI - Short-term morbidity of the upper limb after sentinel lymph node biopsy or axillary lymph node dissection for Stage I or II breast carcinoma. PMID- 14745888 TI - 2003 Warkany Lecture: Autism as a birth defect. PMID- 14745889 TI - Communicating risks during pregnancy: a workshop on the use of data from animal developmental toxicity studies in pregnancy labels for drugs. PMID- 14745890 TI - Sex ratio and associated risk factors for 50 congenital anomaly types: clues for causal heterogeneity. AB - BACKGROUND: Sex ratio (SR) deviations have been reported for many congenital anomalies, but so far no satisfactory explanation for these deviations has been found. The aim of this study was to detect sex-related differences in the association between risk factors and congenital anomalies, and to relate these differences with possibly underlying causes of birth defects. METHODS: Between 1982 and 1999, 1,444,646 newborn infants were examined by the Estudio Colaborativo Latino Americano de Malformaciones Congenitas (ECLAMC) network of South American maternity hospitals. Male relative risks were established for 39,425 infants with 50 selected single anomalies. Associations between male sex and risk factors were identified in nonmalformed infants. In malformed infants, sex-related risk differences were established, and the SR of these infants, with and without associated risk factors, were compared. RESULTS: Infants with neural tube defects (NTDs) and intrauterine growth restriction had a lower SR than those with normal growth, while spina bifida without hydrocephaly (SB[sHy]) was the only NTD subtype without a significant female predominance. Multigravidity lowered the SR of SB(sHy) and HPP (HPP) cases. Increased paternal age inverted the SR of cleft lip (CL) with or without cleft palate (CL[P]) cases from male to female. CONCLUSIONS: The results indicate etiological differences between high and low SB, a stronger relationship between multigravidity and female sex of the offspring than between multigravidity and a specific congenital anomaly, and a possible involvement of dominant mutations for CL(P), as suggested by the association with increased paternal age. PMID- 14745891 TI - Immunostimulation by complete Freund's adjuvant, granulocyte macrophage colony stimulating factor, or interferon-gamma reduces severity of diabetic embryopathy in ICR mice. AB - BACKGROUND: Increased risk of fetal malformation is a complication occurring in pregnant women with type 1 diabetes. Local (uterine) immune stimulation has been shown to reduce diabetes-induced teratogenesis in mice. Limited information is available regarding the ability of diverse methods of maternal immune stimulation to cause this effect or regarding timing requirements of the immune stimulation. METHODS: Diabetes was induced in pregnant ICR mice by streptozocin (STZ) injection. Three different techniques of maternal immune stimulation, complete Freund's adjuvant (CFA), granulocyte-macrophage colony-stimulating factor (GM CSF), or interferon-gamma (IFN-gamma), were then used to stimulate the immune system of the mice. RESULTS: Approximately 50% of fetuses from hyperglycemic (>26 mM/liter blood glucose) dams were malformed, with neural tube defects predominating. Maternal immune stimulation during the time of normoglycemia, i.e., prior to the onset of hyperglycemia, was necessary to reduce teratogenic effects associated with hyperglycemia for each of the immune stimulants. The immune-stimulated diabetic mice then produced significantly lower and approximately equal numbers of malformed fetuses: CFA 20.9%, GM-CSF 23.3%, and IFN-gamma 13.9%. CONCLUSIONS: These results suggest that mechanistically diverse forms of nonspecific immune activation result in protection against diabetes related teratogenesis, but only if given prior to onset of hyperglycemia. PMID- 14745892 TI - Developmental toxicity of arecoline, the major alkaloid in betel nuts, in zebrafish embryos. AB - BACKGROUND: The major alkaloid in the betel nut, arecoline, has been reported to be potent in inducing developmentally toxic effects by generally lowering the embryo weight and retarding development of the embryo. This study examined the adverse effects of arecoline and tried to unravel the mechanism through the tools of molecular biology. METHODS: Arecoline was administered to zebrafish embryos by incubation at concentrations ranging from 0.01-0.04% (wt/vol) and lethality and morphological changes were recorded. The expression of genes was analyzed by reverse transcription-polymerase chain reaction (RT-PCR) and whole-mount in situ hybridization. In addition, the protective effects of several antioxidants were tested. RESULTS: The survival rate of treated embryos during a three-day incubation significantly declined as the arecoline concentration increased. Treated embryos showed general growth retardation and lower rate of heartbeat. When examined at the 24-hr stage, the relative amounts of transcripts of p53, p21, and cyclin D1, and the spatial expression patterns of these genes in treated groups, were comparable to those of the untreated early stages of embryos. Finally, the addition of glutathione (GSH) or its precursor, N-acetyl-L-cysteine (NAC), ameliorated the developmental retardation of embryos by arecoline. CONCLUSIONS: Arecoline-treated embryos exhibited general developmental retardation in a dose-dependent manner. Our results from RT-PCR, in situ hybridization, and antioxidant-protection experiments indicate that the mechanism underlying growth retardation by arecoline in embryos is predominantly due to a general cytotoxic effect induced by depletion of intracellular thiols. PMID- 14745893 TI - Evidence that intrauterine and postnatal androgens affect the development of pyloric stenosis. AB - BACKGROUND: It is rather well established that the causal antecedents of pyloric stenosis (PS) contain both genetic and environmental factors. However, in spite of substantial quantities of epidemiological data, no widespread environmental causal agent has yet been established. There have been recent extensions of our knowledge of the endocrine consequences of fetal growth restriction and of the endocrine determinants of sex ratio. It seemed worth reviewing the epidemiological data on PS in the light of these new developments to see whether this would provide a basis for a plausible hypothesis on these suspected environmental causes of PS. METHODS AND RESULTS: The search terms "pyloric stenosis" and ("epidemiology" or "risk factors") were combined on the Ovid Medline data base for the years 1966 onwards. The data from the resulting papers were augmented by notes occasioned by almost daily reading at the Library of the Royal Society of Medicine (London) for the past 25 years. (Electronic data retrieval systems do not access incidentally recorded data such as sib sex ratios). CONCLUSIONS: It is hypothesized that high intrauterine and early postnatal androgen levels constitute an environmental cause of PS. PMID- 14745894 TI - Mature ovarian cystic teratoma with a highly differentiated homunculus: a case report. AB - BACKGROUND: Mature ovarian cystic teratomas, which are commonly observed benign ovarian tumors, consist of ectodermal, mesodermal, and endodermal components that are generally disorganized. In this report, we document a case in which the solid portion of an ovarian teratoma demonstrated considerable differentiation, forming a doll-like structure. CASE: A 25-year-old virginal Japanese woman underwent surgery for an ovarian tumor that was diagnosed as a mature teratoma. A solid mass within the tumor was found to have a head, trunk, and extremities. Consequently, this mass was diagnosed as a mature fetiform teratoma (homunculus). Brain, eye, spinal nerve, ear, teeth, thyroid gland, bone, bone marrow, gut, trachea, blood vessels, and phallic cavernous tissue were confirmed microscopically. Distinctive features were the clear anterior-posterior, ventral dorsal, and left-right axes, with a spatially well-organized arrangement of the organs. An eye was located on the front of the head, a spinal nerve lay dorsal to the spinal bones, the thyroid gland was anterior to the trachea, and the gut was deep inside the trunk. CONCLUSIONS: These findings indicate that the information necessary for organization of the body plan may be conserved and transmitted, even with parthenogenesis. Mature cystic teratomas of the ovary are mostly benign and do not always attract detailed attention. However, precise analyses of such tumors may significantly enhance our understanding of both parthenogenetic and normal human development. PMID- 14745895 TI - Birth defects among infants of Gulf War veterans, 1989-1993. PMID- 14745913 TI - Differential risks to males and females for congenital malformations among 2.5 million California births, 1989-1997. AB - BACKGROUND: Although many studies have observed variations in the prevalence of specific malformations by sex, there is a lack of population-based data on potential malformation prevalence differences by sex at birth. METHODS: Our objective was to explore differences in the prevalence of structural congenital malformation phenotypes between sexes in a California population of 2.5 million live- and stillbirths, using data from a population-based active surveillance system. Ascertainment was performed among offspring of California women who delivered in nonmilitary hospitals during the period of 1989-1997. Malformations were grouped according to the four-digit malformation codes of the British Pediatric Association. RESULTS: Overall, 32,619 males and 21,835 females were considered to have structural congenital malformations, with prevalences of 2.52% and 1.76%, respectively. Thus, males demonstrated a malformation prevalence that was 22% higher than that in females. Using a criterion of a 40% increase or decrease in the relative risk for males, increased risks for 15 and decreased risks for 17 specific malformation categories were observed. Increased risks were associated with most organ systems, with the notable exception of the nervous system (increased risks for nervous system malformations were observed among female births). Risks were not substantially influenced by adjusting for maternal age, race/ethnicity, parity, or education. CONCLUSIONS: Our observations extend the relatively few studies that have investigated differential prevalences of a large number of specific structural malformations between male and female births. PMID- 14745914 TI - Method of weighted proportion of reproductive-aged women taking folic acid supplements to predict a neural tube defect rate decline. AB - BACKGROUND: Neural tube defect (NTD) rates can be lowered by increased consumption of folic acid (FA) by women before and during early pregnancy. The crude proportion of reproductive-aged women taking FA supplements has been used to predict a decline of the NTD rate in the general population. In this study we examine the potential error in using the crude proportion to predict NTD risk reduction, and offer an alternative method. METHODS: The crude proportion measures the number of women taking FA. It ignores the substantial variability by maternal age in the probability of giving birth. Age-specific fertility rates (ASFRs) reflect the probability that a woman in a specific age group will give birth in a given year. In this study, we show how to calculate a proportion weighted by ASFRs to predict a decline in the NTD rate, and to assess the effectiveness of FA consumption in preventing NTDs. RESULTS: Our results show that a crude proportion of 50% of women (15-49 years old) taking FA is associated with a range of 24-77% in weighted proportions. Assuming a 40% risk reduction from taking 400 microg of FA daily, the expected NTD rate decline could vary from 9.6% to 30.6%, depending on the age distribution of women taking FA. CONCLUSIONS: The ASFR-weighted proportion estimates the proportion of babies born to women taking FA, as opposed to the crude proportion of women taking FA. We recommend using the ASFR-weighted proportion to predict an NTD rate decline and measure the success of FA education campaigns. We found that when women in high-fertility age groups increased their FA consumption, the decline in the NTD rate was greater than when women in low-fertility age groups did so. Our findings suggest that the more efficient approach to NTD prevention is to focus on women with a higher probability of giving birth. For example, by focusing on <50% of women of childbearing age (20-34 years), as much as 76% of the maximum NTD rate reduction can be achieved. PMID- 14745915 TI - First trimester exposure to corticosteroids and oral clefts. AB - BACKGROUND: The possible association between oral cleft in the newborn and maternal exposure to corticoids during pregnancy is still controversial. The aim of this study was to test this association by a case-control analysis using the large multicentric MADRE database. METHODS: The MADRE database is a collection of information on malformed infants with a history of maternal first-trimester drug exposure. Nine malformation registries participate in the data collection. Cases were defined as infants presenting with a cleft palate or cleft lip, and exposure was defined by the use of corticosteroids during the first trimester of pregnancy. RESULTS: After 12 years of data collection, the database includes data on 11,150 malformed infants. A slight association is observed between exposure to corticoids for systemic use and the occurrence of cleft lip with or without cleft palate (OR, 2.59; 95% CI, 1.18-5.67). CONCLUSIONS: If the observed association is real, an interpretation is suggested, based on a likely interaction between corticosteroids and environmental dioxins. It is indeed possible that human fetuses may become sensitive to the teratogenic effect of corticosteroids when they are exposed in utero to environmental pesticides as well. PMID- 14745916 TI - Puerto Rican primary physicians' knowledge about folic acid supplementation for the prevention of neural tube defects. AB - BACKGROUND: We conducted a study of a group of primary physicians in Puerto Rico to evaluate their knowledge about folic acid supplementation to prevent neural tube defects. METHOD: The study design was transverse-correlational. A total of 66 primary physicians in two hospitals (public and private) participated in the study. The sample was nonrandom and opportunistic, and only those physicians present in the hospitals at the moment of distribution of the questionnaires participated. A self-administered and anonymous questionnaire was used. Descriptive statistics and cross-tabular analysis were used to describe the results of this study. Inferential statistics were also used, including Chi square and t-tests to establish the associations/differences between physician knowledge and the independent variables. RESULTS: Of the participants, 87.9% demonstrated an inadequate knowledge about folic acid supplementation for the prevention of neural tube defects as part of preconception care and only 12.1% demonstrated adequate knowledge. Older physicians had greater knowledge about folic acid. Also, women demonstrated greater knowledge about folic acid than did men. Most of the physicians who always recommend supplementation to their patients demonstrated a greater knowledge about folic acid, and all participants with adequate knowledge came from the public hospital. CONCLUSIONS: Despite a concerted effort by the Health Department of Puerto Rico to provide education in the importance of folic acid supplementation to reduce the incidence of neural tube defects, primary physicians in two Puerto Rican hospitals generally have not availed themselves of this training and showed a lack of knowledge on this important clinical issue. PMID- 14745917 TI - Valproate-induced neural tube defects in folate-binding protein-2 (Folbp2) knockout mice. AB - BACKGROUND: Folate is an important B vitamin that is transported into cells by way of folate-binding proteins and transporters. Folate-binding protein-2 nullizygous (Folbp2(-/-)) mice develop normally; however, we have found them to be more susceptible to the teratogenic effects of arsenate exposure than wild type control mice. METHODS: In the current study, we wanted to extend our findings and test the hypothesis that Folbp2(-/-) mice are more susceptible to the teratogenic effects of valproic acid (VPA), a commonly used antiepileptic drug that is known to induce neural tube defects (NTDs) in both humans and laboratory animals. RESULTS: Folbp2(-/-) mice had higher VPA-induced frequencies of embryonic lethality and exencephaly than did the wild-type control mice during folate supplementation and a control diet, respectively. All other differences in response between the two genotypes were short of reaching statistical significance. Folate supplementation of wild-type, but not Folbp2(-/-) dams reduced embryonic lethality of VPA-treated wild-type embryos compared to the folate-deficient diet. CONCLUSIONS: Unlike our previous findings with arsenate, enhanced susceptibility of Folbp2(-/-) mice to in utero VPA exposure was demonstrated in some dietary folate regimens. Thus, our data indicate a relatively frail relationship between Folbp2 and VPA-induced NTDs. PMID- 14745919 TI - New case of non-mosaic tetrasomy 9p in a severely polymalformed newborn girl. AB - BACKGROUND: The phenotypic expression of an additional chromosome 9 causes a very broad clinical spectrum of anomalies. The prognosis for infants with non-mosaic tetrasomy 9p is poor, and they usually die at a very early age. CASE: In this article we present a new case of complete tetrasomy 9p in a newborn girl with multiple dysmorphologic features. Cytogenetic studies were carried out by CBG, GTG, and QFQ chromosome bandings, as well as by fluorescence in situ hybridization (FISH). The cytogenetic findings for the newborn girl showed an extra chromosome interpreted as an isochromosome 9p. The karyotype was characterized as 47,XX,+mar.ish i(9)(p10)(wcp9+). The parental chromosomes were normal. CONCLUSIONS: The karyotype and clinical features of the newborn girl (e.g., typical craniofacial dysmorphism, severe skeletal anomalies, and visceral and genito-urinary malformations), compared with cases reported in the literature, give additional support to a clinical definition of this chromosomal syndrome. PMID- 14745918 TI - Phenotype of the neural tube defect mouse model bent tail is not sensitive to maternal folinic acid, myo-inositol, or zinc supplementation. AB - BACKGROUND: Bent tail is a mouse model for X-linked neural tube defects (NTDs) that is characterized by the presence of exencephaly, a delayed posterior neuropore closure, and a tail phenotype. In addition, Bent tail shows laterality defects and increased prenatal mortality. The congenital malformations of this mouse are caused by a submicroscopic deletion that completely encompasses the gene coding for the zinc finger transcription factor Zic3. In this study we investigated the sensitivity of the phenotype of Bent tail to the nutrients folinic acid, myo-inositol, and zinc. These nutrients are thought to be involved in the etiology of NTDs, in combination with a genetic predisposition. METHODS: The most penetrant phenotype of the Bent tail mouse, the tail malformation, was used as a marker for the nutrient sensitivity of the neural phenotype. The size of the litters and the survival of the offspring, subdivided according to genotype, were analyzed as markers for the nutrient sensitivity of other phenotypic features of Bent tail. RESULTS: In confirmation of earlier studies, we observed the prenatal loss of a number of homozygous females and hemizygous males, as well as the effect of genotype on the tail phenotype of Bent tail. However, periconceptional supplementation of the maternal diet with folinic acid, myo-inositol, or zinc produced no significant effects on either the tail phenotype of the offspring or the size and genotypic composition of the litters. CONCLUSIONS: Bent tail appears to be a folinic acid-, myo-inositol-, and zinc insensitive mouse model for NTDs. PMID- 14745920 TI - Malformations following methimazole exposure in utero: an open issue. AB - BACKGROUND: In hyperthyroidism-complicated pregnancies, medical therapy is necessary to reach an euthyroid condition, and propylthiouracil (PTU) or methimazole (MMI) are used. These drugs are equally effective, but may cause fetal and neonatal hypothyroidism because they freely cross the placenta. Although PTU has not been significantly associated with embryo-fetal anomalies, it has been suggested that MMI might be responsible for a specific embryopathy. CASE(S): Two cases of major congenital anomalies after MMI exposure during pregnancy are reported. CONCLUSIONS: PTU should be the drug of choice, and the use of MMI should be restricted to cases with allergic reactions, intolerance, or poor response to PTU. PMID- 14745921 TI - Multiple congenital anomalies associated with in utero exposure of phenytoin: possible hypoxic ischemic mechanism? AB - BACKGROUND: The characteristics of the phenotype of the malformed phenytoin exposed infant can help to clarify the mechanism of the drug's teratogenesis. One postulated mechanism is vascular disruption. CASE: An infant who was exposed to phenytoin as monotherapy throughout pregnancy was born with the following abnormalities: midface hypoplasia, digit hypoplasia with syndactyly in the hands and feet, meningomyelocele, talipes equinovarus, and a long skin pedicle on the back. The mother was also exposed to cigarette smoking and alcohol during the pregnancy. CONCLUSIONS: The malformations of the hands and feet, and the talipes deformity are potential effects of vascular disruption, a postulated fetal effect of both phenytoin and cigarette smoking. The mechanism of the teratogenicity of phenytoin may have included episodes of bradyarrhythmia in the fetus; however, no such episodes were documented. PMID- 14745922 TI - Topically applied minoxidil may cause fetal malformation: a case report. AB - BACKGROUND: Minoxidil is a K(+) channel opener able to cause relaxation of vascular smooth muscles and modify cell growth and cell fate or migration. It is now widely used for its hair growth promoting effects. When locally applied, it is absorbed through the skin and may have systemic pharmacological effects. CASE: A 28-year-old white pregnant woman daily applied minoxidil 2% to her scalp because of hair loss. At the 22nd gestational week, after a routine ultrasound test showing significant brain, heart, and vascular malformations of the fetus, pregnancy was interrupted. The placenta had numerous ischemic areas and a discrepancy between gestational age and villi maturation. In the villi, capillaries were increased in number, significantly enlarged, and excessively marginalized. The fetus' heart was increased in volume and had a globose shape, the aorta had a distal stenosis. The sigmoid colon was significantly increased in length and a mesentery commune was present. The brain had enlarged ventricles and abundant hemorrhages. Histological examination showed areas of demyelinization with gliosis, signs of excessive and inappropriate angiogenesis, and capillary rearrangement. CONCLUSIONS: Further knowledge on minoxidil-induced fetal toxicity would be beneficial before allowing its use in pregnant women. PMID- 14745923 TI - Mobius sequence in children exposed in utero to misoprostol: neuropathological study of three cases. AB - BACKGROUND: Misoprostol exposure in the first trimester of pregnancy has been related to congenital malformations, particularly the Mobius sequence and terminal transverse limb defects. CASES: Neuropathological findings of three patients with Mobius sequence related to misoprostol are reported. No previous pathological studies have shown these abnormalities to be associated with misoprostol exposure in utero. The brain stem was cut serially, from the rostral mesencephalum to the caudal aspect of the medulla, and all fragments were stained with hematoxylin-eosin and cresyl violet. Old ischemic-anoxic foci of gliosis, with necrosis and calcification, dorsally situated, were present from the pons to the medulla, involving some cranial nerve nuclei (especially the IV, VII, and XII) that were partially or completely depopulated of neural cells. CONCLUSIONS: The findings suggest a circulatory mechanism to the Mobius sequence, with vascular disruption involving the territory of the subclavian artery, occurring in a critical period of embryonic life between six to eight weeks postconception. These cases add further evidence of the role of misoprostol as a teratogen. PMID- 14745924 TI - Statement of the Public Affairs Committee of the Teratology Society on the importance of smoking cessation during pregnancy. PMID- 14745925 TI - A new look at behavioral outcomes and teratogens: a commentary. PMID- 14745926 TI - Folate deficiency is an "imminent health hazard" causing a worldwide birth defects epidemic. PMID- 14745927 TI - Association between maternal fever and psychological/behavior outcomes: a hypothesis. AB - BACKGROUND: This study is one of the first to investigate the association between maternal report of fever during middle to late pregnancy and psychological, behavioral, and educational outcomes in offspring. The hypothesis guiding this research was that maternal fever during the second trimester of pregnancy has an adverse effect on the development of the central nervous system (CNS) of the fetus, resulting in abnormalities of psychological development and behavior that can be observed in childhood. METHODS: Multivariate analyses of a birth cohort compared outcomes for children whose mothers never reported fever during pregnancy and those who reported fever in the second and third trimesters. Children were compared on measures of temperament, behavior, and academic performance in infancy and at five and 12 years of age. RESULTS: Associations were obtained for second-trimester fever and distress to novelty (p < 0.05) in infancy. Significant associations were also obtained for inhibition (p < 0.01), negative emotionality (p < 0.05), and lack of task persistence (p < 0.01) at age five. Furthermore, school achievement (p < 0.05) and task orientation (p < 0.01) at age 12 were associated with maternal reports of second-trimester fever exposure. CONCLUSIONS: Much of the gestation/hyperthermia research has focused on the relationship between hyperthermia exposure and profoundly teratogenic outcomes. In this study we investigated subtler psychological/behavioral associations that may not be observable until later in development. Although the current study was hampered by technical limitations, the results support the need for more rigorously controlled research into a possible association between gestational fever and psychological/behavioral outcomes. PMID- 14745928 TI - Maternal febrile illnesses, medication use, and the risk of congenital renal anomalies. AB - BACKGROUND: Renal anomalies occur in about three infants per 1000 live births and have been associated with several environmental risk factors. Researchers have yet to assess the effect of maternal febrile illnesses on renal anomalies, even though febrile illnesses have been associated with other birth defects. Our objective was to determine whether maternal illness, fever, or medication use during the first trimester of pregnancy is associated with the occurrence of renal anomalies. METHODS: In this population-based case-control study, we evaluated 192 infants with renal anomalies (renal agenesis [n = 44], obstructive defects [n = 134], and renal duplication defects [n = 14]) and 3029 infant without birth defects, all of whom were born in metropolitan Atlanta, Georgia, from 1968 through 1980. Maternal illness was defined as reported flu-like illness and/or episodic illness during the first trimester. RESULTS: Our adjusted multivariate analyses showed that among the 192 case-infants, 38 had mothers with an illness (adjusted odds ratio [AOR], 1.71; 95% confidence interval [CI], 1.15 2.52), 20 had mothers who reported a fever (AOR, 1.80; 95% CI, 1.07-3.02) and 26 had mothers who reported taking medication (AOR, 1.69; 95% CI, 1.07-2.68). Fifteen mothers reported a fever and medication use (AOR, 1.90; 95% CI, 1.05 3.45). Nonprescription aspirin-containing medication use showed the strongest association (AOR, 3.45; 95% CI, 1.36-8.75) with renal anomalies. CONCLUSIONS: Our data suggest that maternal exposure to illness, fever, and medication (particularly aspirin) may increase the risk of congenital renal anomalies. PMID- 14745929 TI - Trend in prevalence of neural tube defects in Quebec. AB - BACKGROUND: In Canada, the first recommendations on the use of folic acid (FA) supplements by women planning a pregnancy or capable of becoming pregnant were issued in 1993. In 1998, fortification of flour with FA became mandatory. The objective of this study was to assess the impact of these measures on the prevalence of neural tube defects (NTDs) in the province of Quebec. METHODS: The study population included stillbirths, live births, and elective terminations for fetal malformations that were reported in 1992-2000 for women residing in the province of Quebec. NTD cases were identified from stillbirth certificates and hospital discharge summaries. RESULTS: There was a marked decrease in the total NTD rate after 1997. The average NTD rate was 1.89 per 1000 total births during the period of 1992-1997, and 1.28 per 1000 in 1998-2000, a 32% reduction (p < 0.001). CONCLUSIONS: Fortification of flour, which began in early 1997 and gradually became widespread, is a very plausible explanation for the timing, shape, and magnitude of the decrease in NTD prevalence observed in Quebec and other Canadian provinces. An increase in FA supplement use may have played only a minor role. Birth Defects Research (Part A) 67:000-000, 2003. PMID- 14745930 TI - Associations between polymorphisms within the thymidylate synthase gene and spina bifida. AB - BACKGROUND: Polymorphisms within the thymidylate synthase (TS) gene that influence enzyme activity may affect plasma folate levels and, indirectly, plasma homocysteine concentrations. We investigated whether TS polymorphisms contribute to spina bifida (SB) risk, given that a reduction in the risk of SB has been linked to folate metabolism. METHODS: Genomic DNA was extracted from newborn screening blood spots obtained from case infants with SB, and randomly selected, nonmalformed control infants. Genotype frequencies of two polymorphisms in the TS gene-a 28-bp tandem repeat in the promoter enhancer region (TSER) and a 6-bp deletion in the 3'UTR-were determined by polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) methods. Additionally, all seven exons of the TS gene were sequenced to identify variations within the coding region of the gene. RESULTS: We found that the TSER 2/2 homozygous genotype was associated with a slightly increased risk for SB infants (odds ratio [OR] = 1.4 [0.8-2.4], p = 0.1). When the cohort was divided into separate ethnic groups, this risk increased by 4-fold with the TSER 2/2 homozygous genotype (OR = 4.0 [1.8-8.8], p = 0.001), and by 3-fold with the 3'UTR +/+ homozygous genotype (OR = 3.6 [1.3-10.1], p = 0.02) in non-Hispanic white cases. The combined TSER,3'UTR (2/2,+/+) genotype showed a more than 4-fold increased risk for SB within this specific ethnic group (OR = 4.7 [1.1-19.8], p = 0.04). CONCLUSIONS: This study is the first to evaluate how TS polymorphisms contribute to the risk of SB. The current findings indicate that polymorphisms in the untranslated regions of the TS gene are associated with 4-fold or more increased risks of SB in non-Hispanic whites, but not in Hispanic whites, African-Americans, or Asian-Americans. PMID- 14745931 TI - Alterations in mitochondrial morphology are associated with hyperthermia-induced apoptosis in early postimplantation mouse embryos. AB - BACKGROUND: Previously, we showed that teratogens such as hyperthermia activate the mitochondrial apoptotic pathway in day nine mouse embryos. Activation of this pathway involves an initial release of cytochrome c from intermembranous spaces of the mitochondria into the cytoplasm. Cytoplasmic cytochrome c then activates a caspase cascade resulting in the orderly demise of the cell. In addition, we showed that teratogens activate the mitochondrial pathway in cells of the neuroepithelium, but not the heart. METHODS: To further investigate the role of the mitochondrion in teratogen-induced apoptosis, we used transmission electron microscopy (TEM) to compare mitochondrial morphology in cells of the neuroepithelium and heart of control and hyperthermia-treated embryos. Because we know that the apoptotic pathway is activated some time during the first 5 hr after teratogen exposure is initiated, we assessed mitochondrial morphology at 1, 2.5, and 5 hr after day nine mouse embryos were exposed to hyperthermia (43 degrees C, 15 min). RESULTS: In neuroepithelial cells of the prosencephalon, abnormally-shaped mitochondria were observed at the 1 hr time point and thereafter, whereas loss of cristae and shrunken mitochondria were noted at the 5 hr time point. In contrast, no obvious changes in mitochondria of heart cells were observed at any of the time points monitored. CONCLUSIONS: These results indicate that teratogen-induced cell death in neuroepithelial cells is temporally correlated with alterations in mitochondrial morphology, whereas the absence of cell death in the heart is correlated with a corresponding lack of change in mitochondrial morphology. Birth Defects Research (Part A), 2003. PMID- 14745932 TI - Timetable for intestinal rotation in staged human embryos and fetuses. AB - BACKGROUND: The existing data on intestinal rotation during human development are contradictory regarding the timing of major events, and as such an exact timetable for rotation of the intestine in humans is not yet available. METHODS: We studied the initial formation and rotation of the intestine by microdissection and histological observations in 72 human embryos and fetuses at two to 12 weeks postfertilization. The embryos were classified according to the Carnegie staging system. RESULTS: The primordium of the primitive gut was first observed as a yolk sac at stage 5. With the formation of the embryonic foldings, three divisions of the primitive gut (the foregut, midgut, and hindgut) were observed at stage 10. At stage 12, the primitive gut was located on the midline. At stage 15, a 90 degrees counterclockwise rotation of the intestine began. At stage 16, herniation of the intestine into the umbilical cord was not evident in observations of the external form or a transversely sectioned embryo, but was evident in a sagittally sectioned embryo. There was another 90 degrees counterclockwise rotation at stage 20. Reduction of the intestine was a rapid process, since it was still in the cord in fetuses of <40 mm crown-rump length (CRL), and was reduced above 40 mm in general during nine weeks of development. When the intestine returned to the abdominal cavity, the cecum was located in the right lower quadrant (the adult position). CONCLUSIONS: We have developed a standard timetable to describe the rotation of the intestine. The current results will be helpful in studies describing the pathogenesis of some developmental abnormalities in the intestine due to abnormal rotation. PMID- 14745933 TI - Preimplantation genetic diagnosis for a couple with recurrent pregnancy loss and triploidy. AB - BACKGROUND: Triploidy may arise from fertilization of a mature haploid egg by two haploid sperm or by failure of meiotic divisions yielding a diploid gamete. We encountered a couple with habitual abortion, in which the last two fetuses were documented as viable triploid. METHODS: To avoid dispermic penetration and development of abnormal preembryos, insemination was done by intracytoplasmic sperm injection (ICSI) followed by fluorescence in situ hybridization (FISH) of biopsied blastomeres. RESULTS: Tests of the husband's spermatozoa by FISH, revealed that only 2-3% of the sperm were disomic for chromosomes 16, 13, 21, X, and Y. No triple disomy was detected among chromosomes 16, 13 and 21, which makes it very unlikely that triploidy resulted from diploid spermatozoa. Following a controlled ovulation induction protocol, low quality oocytes with immature cumuli were revealed. After ICSI, five eggs became two pronuclei (2PN) zygotes and none of the other eggs developed a 3PN zygote. FISH was performed on chromosomes 16 and 21 in four preembryos developed to a 6-8 cell stage. Aneuploidy or mosaicism for each of these chromosomes was detected in one preembryo and later in two disaggregated blastocysts. FISH failed in one preembryo that became atretic after biopsy. CONCLUSIONS: Although this case was unsuccessful in achieving embryo transfer and normal pregnancy, we detected many abnormal morphological features in the oocytes and chromosomal abnormalities in the cleaving preembryos. This protocol can be proposed to patients with recurrent pregnancy loss associated with chromosomal abnormalities in the fetus. PMID- 14745934 TI - Fetal bilateral renal agenesis, phocomelia, and single umbilical artery associated with cocaine abuse in early pregnancy. AB - BACKGROUND: Maternal cocaine abuse in pregnancy is associated with complications such as intrauterine growth retardation, abruptio placentae, and preterm delivery. CASE: We report what is, to our knowledge, the first published observation of fetal bilateral renal agenesis associated with a vascular disruption syndrome comprising upper limb reduction defect and a single umbilical artery following maternal cocaine abuse in early pregnancy. CONCLUSIONS: This constellation in a fetus aborted at 18 weeks extends the spectrum of complications possibly associated with cocaine abuse in pregnancy. PMID- 14745936 TI - Molecular epidemiology of hypospadias: review of genetic and environmental risk factors. AB - Hypospadias is one of the most common congenital anomalies in the United States, occurring in approximately 1 in 125 live male births. It is characterized by altered development of the urethra, foreskin, and ventral surface of the penis. In this review, the embryology, epidemiology, risk factors, genetic predisposition, and likely candidate genes for hypospadias are described. Recent reports have identified increases in the birth prevalence of mild and severe forms of hypospadias in the United States from the 1960s to the present. Studies in consanguineous families and small case series have identified allelic variants in genes controlling androgen action and metabolism that cause hypospadias, but the relevance of these findings to the general population is unknown. Concern has also focused on whether exposure to endocrine disrupting chemicals (EDC) with antiandrogenic activity is the cause of this increase. Hypospadias is believed to have a multifactorial etiology in which allelic variants in genes controlling androgen action and metabolism predispose individuals to develop this condition. When genetic susceptibility is combined with exposure to antiandrogenic agents, a threshold is surpassed, resulting in the manifestation of this birth defect. A clear role for exposure to antiandrogenic environmental chemicals has yet to be established in the etiology of hypospadias, although results from laboratory animal models indicate that a number of environmental chemicals could be implicated. Molecular epidemiology studies that simultaneously examine the roles of allelic variants in genes controlling androgen action and metabolism, and environmental exposures are needed to elucidate the risk factors for these anomalies and the causes of the increased rate of hypospadias. PMID- 14745937 TI - Antibodies to folate receptors impair embryogenesis and fetal development in the rat. AB - BACKGROUND: Folic acid (FA) supplementation reduces neural tube defects (NTDs) by 70%. However, the cause of most NTDs cannot be attributed to folate deficiency, to mutations of genes that encode folate pathway enzymes, and folate receptors (FRs) that mediate cellular folate uptake. Mouse embryos nullizygous for the ortholog of the FRalpha gene have lethal congenital abnormalities that are preventable by administration of folinic acid to the dams. To determine whether antibodies to FRs are similarly teratogenic, we studied a rat model. METHODS: Immunohistochemistry with an antiserum to rat FRs was used to identify the receptors on reproductive tissues and embryos. Gestation day (GD) 8 rats received intraperitoneal injections of antiserum to the FRs, and their embryos were examined 2-9 days later. Some rats received pharmacologic doses of folinic acid or dexamethasone before the antiserum was administered. RESULTS: The FRs are present on oocytes, the oviduct, and uterine epithelial cells, and in the embryo at all stages examined between GD4 and GD15. The antiserum has a dose-related effect on embryo viability and organogenesis. Folinic acid prevented teratogenicity resulting from smaller doses of antiserum, but not that caused by larger doses. Resorption of embryos with the larger doses of the antiserum was prevented by dexamethasone. CONCLUSIONS: FRs are expressed on oocytes, epithelial cells of reproductive organs, and embryonic and extraembryonic tissues. Antiserum to FRs administered to pregnant rats causes embryonic damage. Embryo lethality with smaller doses of antiserum is preventable by administration of folinic acid, while larger doses cause embryo damage by immune-mediated cell lysis, which can be prevented by dexamethasone. PMID- 14745938 TI - Caspase-3 mediates retinoid-induced apoptosis in the organogenesis-stage mouse limb. AB - BACKGROUND: Caspases are key mediators in the regulation and execution of apoptosis, a crucial part of the morphogenetic process during limb development. Caspase-8 and -9 are upstream caspases. Caspase-8 mediates the extrinsic pathway of apoptosis triggered by signaling through TNF-R1 family receptors. Caspase-9 is activated during the intrinsic pathway downstream of mitochondria. Caspase-3 is an effector caspase that initiates degradation of the cell in the final stages of apoptosis. Vitamin A is a potent teratogen that causes limb reduction defects in embryos exposed during organogenesis. Previous in vitro studies have shown that exposure of the organogenesis-stage murine limb to vitamin A results in excessive levels of apoptosis. The goal of this work was to characterize the involvement of caspase-3, -8, and -9, as well as cytochrome-c release from the mitochondria, in the apoptotic cascade induced by vitamin A. METHODS: Limb buds from gestational day 12 CD-1 mice were cultured in a chemically defined medium in the absence or presence of vitamin A. Cultures were terminated after 6 days to examine the effect of the drug on gross morphology. Apoptosis was detected by TUNEL staining after culture for 24 hr. Caspase activation was determined by Western blotting and localized by immunohistochemistry of control and treated limbs. The release of cytochrome-c into the cytoplasm was assessed by Western blotting after cell fractionation. RESULTS: Limbs cultured in the presence of vitamin A showed a dose dependent growth reduction and dysmorphogenesis of the cartilaginous anlagen. Apoptosis was increased in the interdigital, anterior, and posterior marginal zones and in the apical ectodermal ridge. Western-blotting confirmed the presence of activated caspase-3 that increased with time in culture and vitamin A concentration. Cleaved caspase-3 immunoreactivity colocalized with TUNEL stained limb regions and increased dramatically with increasing drug concentrations. In contrast, procaspase-8 and -9 were not activated. Exposure to high concentrations of vitamin A did, however, increase cytoplasmic cytochrome-c, suggesting mitochondrial involvement. CONCLUSIONS: Caspase-3 is a key effector caspase in the apoptotic pathway induced by Vitamin A. While caspases-8 and -9 are not responsible for the activation of caspase-3 in response to the drug, cytochrome-c release from mitochondria may play an upstream role. PMID- 14745939 TI - Developmental immunotoxicity of lead: impact on thymic function. AB - BACKGROUND: Since the potential effects of early exposure to lead on thymic functions have not been fully characterized, in this study we evaluated the capacity of lead to alter thymic function in juvenile chickens following embryonic exposure. METHODS: Cornell K strain White Leghorn chicken eggs were administered lead acetate (400 microg/egg) or sodium acetate (control) on embryonic development (E12) with and without thymulin supplementation. Ex vivo production of interferon-gamma (IFN-gamma)-like cytokine by thymocytes and a delayed-type hypersensitivity (DTH) reaction were measured in the juvenile. Additionally, the effects of in vitro exposure to lead on both thymocytes and thymic stromal cells (TSCs) were evaluated. RESULTS: Following E12 exposure to lead, ex vivo production of IFN-gamma-like cytokine by juvenile-derived thymocytes decreased significantly compared to the control. The same effect was observed when thymocytes were directly exposed to lead in vitro and stimulated with thymic stromal supernatant. In contrast, when TSCs were exposed to lead in vitro, no change was seen in their functional capacity for promoting cytokine production. In ovo supplementation with thymulin partially reversed lead-induced DTH depression without any change in IFN-gamma-like cytokine production. Embryonic exposure to thymulin alone partially depressed the DTH response. CONCLUSIONS: These results suggest that lead can directly influence thymocyte function in the absence of the thymic microenvironment. Since thymulin levels may influence lead-induced immunotoxicity, embryonic endocrine status may be an important consideration. Lead exposure appears to alter thymic functions directly; however, indirect effects via endocrine factors are not precluded. PMID- 14745940 TI - Persistent Mullerian duct syndrome caused by both a 27-bp deletion and a novel splice mutation in the MIS type II receptor gene. AB - BACKGROUND: Persistent Mullerian duct syndrome (PMDS) is a rare form of male pseudohermaphroditism that is characterized by the persistence of Mullerian derivatives in otherwise normally virilized males. Mutations of the Mullerian inhibiting substance (MIS) gene or the MIS type II receptor (MISRII) gene have been identified in PMDS patients with autosomal recessive transmission. We analyzed a compound heterozygote PMDS patient who had a 27-bp deletion in exon 10 in one allele and a novel mutation in intron 5 in the other allele of the MISRII gene. METHODS: Whole blood and tissue samples were obtained from a one-month-old 46,XY male with persistent PMDS and the MISRII gene was sequenced and compared to his mother's genomic DNA and that of 22 normal individuals. Serum MIS and the reproductive hormones were measured by standard immunoassays. RESULTS: The patient's hormone levels were normal but the gene for MISRII contained several mutations, a 27-bp deletion in exon 10 on one allele (one of the most common mutations in PMDS) and a novel mutation in intron 5 in the other allele that altered splicing, resulting in retention of the intron and a frameshift, introducing a stop codon. Other mutations in introns 6 and 9 and in exon 11 might not be functionally significant. CONCLUSIONS: This case reveals a novel mutation in the MISRII gene involving intronic sequences, which when coexisting with the already identified 27-bp deletion in exon 10, leads to PMDS. PMID- 14745941 TI - Maternal periconceptional alcohol consumption and risk for conotruncal heart defects. AB - BACKGROUND: In this study we investigated whether the risk of delivering infants with conotruncal heart defects was increased among mothers who consumed alcohol during the periconceptional period (i.e., 1 month before conception to 3 months after conception). METHODS: Data were obtained from a population-based case control study of California births from 1987-1988. Information concerning alcohol consumption was obtained via telephone interviews with mothers of 207 (87% of eligibles) case infants and 481 (76%) nonmalformed control infants. RESULTS: Bivariate results indicated that relative to nonconsumers, women who consumed alcohol less than once a week had a 1.3-fold increased risk of delivering infants with a conotruncal heart defect (95% confidence interval (CI) 1.0, 1.9), and women who consumed alcohol once a week or more had a 1.9-fold increased risk (95% CI 1.0, 3.4). The risks associated with consuming five or more drinks per drinking occasion were 1.6 (95% CI 0.8, 3.2) for less than once a week, and 2.4 for once a week or more (95% CI 0.6, 9.7). The results for the phenotypic subgroups were similar to those for all cases. Adjustment for potential covariates resulted in somewhat weaker, but still elevated, risks. CONCLUSIONS: This study found that the risk of conotruncal heart defects in offspring was moderately elevated among women who consumed alcoholic beverages during the periconceptional period, and that risk was higher with increased frequency of drinking or increased number of drinks consumed per occasion. Most of the risk estimates were imprecise, and chance could not be ruled out as an explanation for the observed findings. PMID- 14745942 TI - Increased risk of birth defects among children from multiple births. AB - BACKGROUND: Multiple births are increasing, and may be associated with birth defects. METHODS: To explore this relationship, data from the Virginia birth defects registry (VaCARES) was analyzed. RESULTS: During 1989-1998, a total of 44505 children from singleton births and 2258 children from multiple births were born with birth defects in Virginia. The risk of birth defects was significantly increased in children from multiple births as compared to singleton births (birth defect rate per 10000 live births: singleton 482.3, twin 922.0, triplet 1300.0, and quadruplet or higher 2222.2). Increased risk was observed for 39 of 86 diagnoses. The five diagnoses with the highest risk ratio per 10000 live births (RR) were: neurofibromatosis (RR, 12.80), retrolental fibroplasia (RR, 9.96), microphthalmos (RR, 5.24), pulmonary valve anomalies (RR, 5.00), and patent ductus arteriosus (RR, 4.68). A significantly reduced risk ratio for congenital hip dislocation was found in these children (RR, 0.54). In most multiple births, only one child was born with birth defects (81% in twin births, 71% in triplet births, and 56% in quadruplet and higher births). The diagnosis of birth defects might be concordant or discordant for children in which all siblings had birth defects. The children from multiple births who had birth defects were generally preterm and had significantly lower birth weight. A higher fatality and mortality rate and longer hospital stay were also observed. CONCLUSIONS: Overall, children from multiple births have an increased risk of birth defects. PMID- 14745943 TI - Pediatric neurobehavioral diseases in Nevada counties with respect to perchlorate in drinking water: an ecological inquiry. AB - BACKGROUND: Contamination of drinking water with perchlorate, a known thyrotropic agent, has been demonstrated in areas in the western United States. The health consequences of that exposure have been studied, particularly in the State of Nevada. Previous studies in Nevada, comparing the area with perchlorate in the drinking water and the areas without perchlorate in the drinking water, have found no difference in neonatal thyroxine (T(4)) or thyrotropin (TSH) levels, or in the prevalences of thyroid diseases and thyroid cancer. This same study design has now been applied to the major neurobehavioral diseases of childhood (i.e., attention deficit-hyperactivity disorder (ADHD) and autism) and to school performance in order to determine whether those conditions are more frequent in the area with perchlorate-contaminated water. METHODS: Medical services data on ADHD and autism were obtained from the Nevada Medicaid system for the period of January 1, 1996, to December 31, 2000, with county of residence used as the basis for residential information. Analyses of fourth-grade school performance results for two recent time periods came from the state government. Perchlorate concentrations in drinking water had been determined by local water authorities. ADHD and autism rates for the area with perchlorate in the drinking water (Clark County) were calculated and compared with the rates for the other areas in the state, as were fourth-grade school performances. RESULTS: Analysis of the data from the Nevada Medicaid program shows that the rates for ADHD and for autism in the area where perchlorate was in the drinking water did not exceed the rates in those areas where there was no perchlorate contamination in the drinking water. Fourth-grade standardized test results for students in Clark County were not different from those of the remainder of the state. CONCLUSIONS: This ecological study of children in the exposure area did not find evidence of an increased risk of either ADHD or of autism caused by perchlorate contamination in the drinking water. Furthermore, no difference in overall fourth-grade school performance was observed. No evidence was found that children from the area with perchlorate in the drinking water (up to 24 microg/liter) had either an increase in pediatric neurobehavioral disease (ADHD and autism) or a decrease in fourth-grade academic performance. The limitations of this ecological study relate to diagnostic criteria and ascertainment of geographic and demographic differences and to data on individual residence and water consumption during pregnancy. PMID- 14745948 TI - DAN directs endolymphatic sac and duct outgrowth in the avian inner ear. AB - Bone morphogenetic proteins (BMPs) are expressed in the developing vertebrate inner ear and participate in inner ear axial patterning and the development of its sensory epithelium. BMP antagonists, such as noggin, chordin, gremlin, cerberus, and DAN (differential screening-selected gene aberrative in neuroblastoma) inhibit BMP activity and establish morphogenetic gradients during the patterning of many developing tissues and organs. In this study, the role of the BMP antagonist DAN in inner ear development was investigated. DAN-expressing cell pellets were implanted into the otocyst and the periotic mesenchyme to determine the effects of exogenous DAN on otic development. Similar to the effects on the inner ear seen after exposure of otocysts to the BMP4 antagonist noggin, semicircular canals were truncated or eliminated based upon the site of pellet implantation. Unique to the DAN implantations, however, were effects on the developing endolymphatic duct and sac. In DAN-treated inner ears, endolymphatic ducts and sacs were merged with the crus or grew into the superior semicircular canal. Both the canal and endolymphatic duct and sac effects were rescued by joint implantation of BMP4-expressing cells. Electroporation of DAN antisense morpholinos into the epithelium of stage 15-17 otocysts, blocking DAN protein synthesis, resulted in enlarged endolymphatic ducts and sacs as well as smaller semicircular canals in some cases. Taken together, these data suggest a role for DAN both in helping to regulate BMP activity spatially and temporally and in patterning and partitioning of the medial otic tissue between the endolymphatic duct/sac and medially derived inner ear structures. PMID- 14745949 TI - Calcium transients regulate patterned actin assembly during myofibrillogenesis. AB - The highly ordered arrangement of sarcomeric myosin during striated muscle development requires spontaneous calcium (Ca(2+)) transients. Here, we show that blocking transients also compromises patterned assembly of actin thin filaments, titin, and capZ. Because a conserved temporal assembly pattern has been described for these proteins, selective inhibitors of either thick or thin filament formation were used to determine their relative temporal interdependencies. For example, inhibition of myosin light chain kinase (MLCK) by application of a specific inhibitory peptide or phorbol myistate acetate (PMA) disrupts myosin assembly without significantly affecting formation of actin bands. The MLCK inhibitor ML-7, however, disrupted actin as well as myosin. Surprisingly, agents that interfere with actin dynamics, such as cytochalasin D, produced only minor organizational disruptions in actin, capZ, and titin staining. However, cytochalasin D and other actin disrupting compounds significantly perturbed myosin organization. The results indicate that (1) Ca(2+) transients regulate one or more of the earliest steps in sarcomere formation, (2) mature actin filaments can assemble independently of myosin band formation, and (3) myosin thick filament assembly is extremely sensitive to disruption of either the actin or titin filament systems. PMID- 14745950 TI - Properties of four human embryonic stem cell lines maintained in a feeder-free culture system. AB - Several laboratories have begun evaluating human ES (hES) cell lines; however, direct comparisons between different hES cell lines have not been performed. We have characterized the properties of four human cell lines maintained in feeder free culture conditions. Quantitative assessment of surface markers, microarray analysis of gene expression patterns, expression of SOX-2, UTF-1, Rex-1, OCT3/4, CRIPTO, and telomerase activity demonstrated similar patterns in all hES cell lines examined. Undifferentiated hES cells do not respond to neurotransmitters such as acetylcholine, glutamate, and gamma-aminobutyric acid. In addition, the undifferentiated hES cells possess gap junctions. Although similarities in marker expression were observed, allotyping showed that all four lines have a distinct HLA profile, predicting differences in transplantation responses. These data provide the first detailed comparison of different hES cell lines and demonstrate remarkable similarities among lines maintained in identical culture conditions. PMID- 14745951 TI - Long-term culture of human embryonic stem cells in feeder-free conditions. AB - We have demonstrated previously that human embryonic stem (hES) cells possess a characteristic morphologic, antigenic, and molecular profile that can be used to assess the state of ES cells (Carpenter et al., [2004] Dev Dyn 229:243-258). In this manuscript, we have examined the long-term stability of three hES cell lines in feeder-free culture. We demonstrate that the expression of antigens and transcription factors, telomerase activity, telomere length, and karyotype appear stable for all three hES cell lines after continuous culture for over 1 yr. All three lines retained pluripotent differentiation in vitro and in vivo. Although hES cell lines were remarkably stable over the period of analysis, a detailed quantitative analysis of antigen expression by flow cytometry and gene expression by microarray suggested that cell lines show subtle differences in the expression of small subsets of genes upon long-term culture. PMID- 14745952 TI - Amphibian cardiac troponin I gene's organization, developmental expression, and regulatory properties are different from its mammalian homologue. AB - In mammals, the expression of the troponin I-slow (TnIs) isoform is predominant in the heart during embryogenesis and, shortly after birth, is replaced by the cardiac-specific isoform, TnIc; a developmental switch thought to be mediated by thyroid hormone. Whereas, in Xenopus, TnIc is expressed at the onset of heart formation and is the only TnI isoform expressed in the heart. Herein, we demonstrate that the expression patterns of these genes appear to be common within the anuran lineage and, unlike their mammalian counterparts, are not affected by thyroid hormone. To elucidate the regulatory mechanism(s) governing the expression of the amphibian TnIc gene, we characterized the TnIc gene from Rana catesbeiana and used its 5'-flanking region to drive expression of green fluorescent protein in the Xenopus transgenic system. Our results demonstrate that a 300-bp minimal promoter containing intact GATA and CArG-box elements is sufficient to drive expression of this reporter gene in a pattern that mimics, both spatially and temporally, the expression of the endogenous Xenopus TnIc gene. PMID- 14745953 TI - Pilot morpholino screen in Xenopus tropicalis identifies a novel gene involved in head development. AB - The diploid frog X. tropicalis has recently been adopted as a model genetic system, but loss-of-function screens in Xenopus have not yet been performed. We have undertaken a pilot functional knockdown screen in X. tropicalis for genes involved in nervous system development by injecting antisense morpholino (MO) oligos directed against X. tropicalis mRNAs. Twenty-six genes with primary expression in the nervous system were selected as targets based on an expression screen previously conducted in X. laevis. Reproducible phenotypes were observed for six and for four of these, a second MO gave a similar result. One of these genes encodes a novel protein with previously unknown function. Knocking down this gene, designated pinhead, results in severe microcephaly, whereas, overexpression results in macrocephaly. Together with the early embryonic expression in the anterior neural plate, these data indicate that pinhead is a novel gene involved in controlling head development. PMID- 14745954 TI - Calcineurin signaling in avian cardiovascular development. AB - Experiments were initiated in avian embryos to determine the embryonic expression of calcineurin protein phosphatase isoforms as well as to identify developmental processes affected by inhibition of calcineurin signal transduction. Chicken calcineurin A alpha (CnAalpha) and calcineurin A beta (CnAbeta) are differentially expressed in the developing cardiovascular system, including primitive heart tube and valve primordia. Inhibition of calcineurin signaling by cyclosporin A (CsA) treatment in ovo resulted in distinct cardiovascular malformations, depending on the timing and localization of treatment. Initial formation of the heart tube was apparently normal in embryos treated with CsA from embryonic day (E)1 to E2, but hallmarks of heart failure were apparent with treatment from E2 to E3. Vascular defects were apparent in whole embryos treated on either day, but local administration of CsA directly to the forming vessels on E2 did not inhibit blood vessel formation. This observation supports an indirect effect of calcineurin inhibition on angiogenic remodeling as a result of compromised heart development. Together these studies are consistent with multiple roles for calcineurin signaling in the developing cardiovascular system. PMID- 14745955 TI - Endothelium-specific Cre recombinase activity in flk-1-Cre transgenic mice. AB - The use of the Cre-loxP recombination system allows the conditional inactivation of genes in mice. The availability of transgenic mice in which the Cre recombinase expression is highly cell type specific is a prerequisite to successfully use this system. We previously have characterized regulatory regions of the mouse flk-1 gene sufficient for endothelial cell-specific expression of the LacZ reporter gene in transgenic mice. These regions were fused to the Cre recombinase gene, and transgenic mouse lines were generated. In the resulting flk 1-Cre transgenic mice, specificity of Cre activity was determined by cross breeding with the reporter mouse lines Rosa26R or CAG-CAT-LacZ. We examined double-transgenic mice at different stages of embryonic development (E9.5-E16.5) and organs of adult animals by LacZ staining. Strong endothelium-specific staining of most vascular beds was observed in embryos older than E11.5 in one or E13.5 in a second line. In addition, the neovasculature of experimental BFS-1 tumors expressed the transgene. These lines will be valuable for the conditional inactivation of floxed target genes in endothelial cells of the embryonic vascular system. PMID- 14745956 TI - Differentiation trapping screen in live culture for genes expressed in cardiovascular lineages. AB - We have developed a gene trap vector that transduces an EGFP-neo fusion gene (Eno) to monitor the expression of trapped genes in living cells and embryos. Upon in vitro differentiation, most gene-trapped embryonic stem (ES) cell clones exhibited detectable green fluorescence in various specialized cell types, which can be followed in the live culture in real time. Populations of ES cell-derived cardiomyocytes, smooth muscle cells, vascular endothelial cells, and hematopoietic cells were readily recognized by their distinctive morphologies coupled with unique activities, allowing efficient screening for clones with trapped genes expressed in cardiovascular lineages. Applying G418 selection in parallel differentiation cultures further increased detection sensitivity and screening throughput by enriching reporter-expressing cells with intensified green fluorescent protein signals. Sequence analyses and chimera studies demonstrated that the expression of trapped genes in vivo closely correlated with the observed lineage specificity in vitro. This provides a strategy to identify and mutate genes expressed in lineages of interest for further functional studies. PMID- 14745957 TI - Differences in the embryonic expression patterns of mouse Foxf1 and -2 match their distinct mutant phenotypes. AB - Murine genes encoding the forkhead transcription factors Foxf1 and -2 are both expressed in derivatives of the splanchnic mesoderm, i.e., the mesenchyme of organs derived from the primitive gut. In addition, Foxf2 is also expressed in limbs and the central nervous system. Targeted mutagenesis of Foxf1 and -2 suggests that Foxf1 is the more important of the two mammalian FoxF genes with early embryonic lethality of null embryos and a haploinsufficiency phenotype affecting foregut-derived organs. In contrast, the only reported defect in Foxf2 null embryos is cleft palate. To investigate if the differences in mutant phenotype can be attributed to nonoverlapping expression patterns or if distinct functions of the encoded proteins have to be inferred, we analyzed the early embryonic expression of Foxf2 and compared it with that of the better investigated Foxf1. We find that in the early embryo, Foxf1 is completely dominating-in terms of expression-in extraembryonic and lateral plate mesoderm, consistent with the malformations and early lethality of Foxf1 null mutants. Along the developing gut, Foxf1 is highly expressed throughout, whereas Foxf2 expression is concentrated to the posterior part-fitting the foregut haploinsufficiency phenotypes of Foxf1 mutants. Foxf2, on the other hand, is more prominent than Foxf1 in mesenchyme around the oral cavity, as would be predicted from the cleft palate phenotype. The differences in expression pattern also highlight areas where defects should be sought for in the Foxf2 mutant, for example limbs, the posterior gut, genitalia, and derivatives of the neural crest mesenchyme. PMID- 14745958 TI - Isolation and expression of the homeobox gene Gbx1 during mouse development. AB - In zebrafish, gbx1 and otx2 are among the earliest genes expressed in the neuroectoderm, dividing it into an anterior and a posterior domain with a common border that marks the midbrain-hindbrain boundary (MHB) primordium. Here, we describe the sequence and expression pattern of Gbx1 in mouse. The first transcripts are found at embryonic day 7.75 in the hindbrain. Later on, expression of Gbx1 is detectable in the hindbrain (rhombomeres 2 to 7), spinal cord, optic vesicles, and in the ventral telencephalon. In mouse, Gbx1 expression is not observed at the MHB as is the case during early zebrafish development. We suggest that an evolutionary switch occurred: in mouse Gbx2 is involved in the early specification of the MHB primordium, whereas in zebrafish, gbx1 is required instead of gbx2. PMID- 14745959 TI - Zinc finger protein Zac1 is expressed in chondrogenic sites of the mouse. AB - Zac1 is a zinc finger transcription factor that elicits antiproliferative activity and is a potential tumor suppressor gene. Through a detailed spatiotemporal study by in situ hybridization of mouse embryos, we have found that Zac1 transcript is predominantly localized in developing chondrogenic tissue, in addition to the central nervous system as reported elsewhere. Zac1 is also expressed transiently in the myocardium, skeletal muscle, and basal aspect of the stratified embryonic epithelia. During cartilage development, the pattern of Zac1 expression is in close accordance with the distribution of type II collagen mRNA in mesenchymal condensation and prehypertrophic chondrocytes. In mouse ATDC5 cells undergoing in vitro chondrogenesis, the Zac1 mRNA is up regulated in parallel with genes expressed in precartilage but the Zac1 expression is low when type II collagen mRNA is markedly increased in differentiated cells. Together, these results suggest that Zac1 is a potential regulatory gene involved in chondrogenic differentiation. PMID- 14745960 TI - Identification of the mammary line in mouse by Wnt10b expression. AB - Mammogenesis in rabbit, rat, and human begins with the formation of an elevated ectodermal ridge in the embryo. Attempts to demonstrate a morphologically or histologically equivalent mammary line in the mouse have yielded controversial results. We show here that a mammary line exists in the mouse embryo at embryonic day (E) 11.25 as a concise line of Wnt10b expression and a broader band of Wnt6 expression in the surface ectoderm, between the subaxillary and suprainguinal region of each flank. Additional streaks of Wnt10b expression in the axillary and inguinal region join the mammary line on the flank slightly later. Expression of Wnt10b and Wnt6 becomes restricted to the placodes within 1.5 days. The ectoderm of the mammary line is organized as a pseudostratified epithelium connecting the developing mammary placodes at around E11.5, whereas all other surface ectoderm is single-layered. Thus, the mammary line expressing Wnt10b defines a distinct ectodermal region that we consider the equivalent of the ectodermal ridge in, for example, rabbit. To date, the formation of the mammary line expressing Wnt10b is the earliest discernible ectodermal event in murine embryonic mammary gland development. PMID- 14745961 TI - Survey of forkhead domain encoding genes in the Drosophila genome: Classification and embryonic expression patterns. AB - Genetic approaches in Drosophila led to the identification of Forkhead, the prototype of forkhead domain transcription factors that are now known to comprise an evolutionarily conserved family of proteins with essential roles in development and differentiation. Sequence analysis of the recently published genomic scaffold sequence from Drosophila melanogaster has allowed us to determine the presumably full complement of forkhead domain encoding genes in this species. We show herein that the Drosophila genome contains 17 forkhead domain encoding genes; 13 of these genes have orthologs in chordate species, and their products can be assigned to 10 of the 17 forkhead domain subclasses known from chordates. One Drosophila forkhead domain gene only has a Caenorhabditis elegans ortholog and may represent a subclass that is absent in chordates, while the remaining three cannot be classified. We present the mRNA expression patterns of seven previously uncharacterized members of this gene family and show that they are expressed in tissues from all three germ layers, including central and peripheral nervous system, epidermis, salivary gland primordia, endoderm, somatic mesoderm, and hemocyte progenitors. Furthermore, the expression patterns of two of these genes, fd19B and fd102C, suggest a role for them as gap genes during early embryonic head segmentation. PMID- 14745962 TI - Tracing of Xenopus tropicalis germ plasm and presumptive primordial germ cells with the Xenopus tropicalis DAZ-like gene. AB - A gamete is derived initially from a presumptive primordial germ cell (pPGC) and transmits genetic potential to the next generation. Xenopus tropicalis, which is a close relative of Xenopus laevis, has a diploid genome and advantages for genetic and genomic research; however, little is known about the developmental mechanism of its germinal lineage. Here, we identified the Xenopus tropicalis DAZ like gene (Xtdazl), which encodes RNA-binding proteins homologous to Xdazl in Xenopus laevis and examined the expression patterns of Xtdazl transcripts during embryogenesis. In this work, we showed that Xtdazl mRNA was localized in the germ plasm and was expressed from the previtellogenic oocyte to early tadpole, in testis and ovary. The same localization patterns have been reported in Xenopus laevis germ plasm and pPGCs. These results indicate that Xtdazl mRNA is the first specific marker of germ plasm and pPGCs in Xenopus tropicalis and is very useful to trace Xenopus tropicalis pPGCs, including germ plasm until the early tadpole stage. PMID- 14745963 TI - Identification of a novel left-right asymmetrically expressed gene in the mouse belonging to the BPI/PLUNC superfamily. AB - In the process of left-right (L-R) axis formation in the mouse, the node plays a critical role as a structure where the initial breaking of L-R symmetry occurs. Here, we report on the gene LPlunc1, a member of BPI/PLUNC gene superfamily, which is asymmetrically expressed in the developing mouse node. LPlunc1 protein is secreted as a processed form of relative molecular mass 54K-60K and shares sequence features with the other members of BPI/PLUNC superfamily, including the N-terminal and C-terminal homology domains, each of which is considered to form a lipid binding pocket. LPlunc1 is transiently expressed in the crown cells of the node asymmetrically. This expression pattern of Lplunc1 highly overlaps with that of Nodal, a major player during the L-R formation. Interestingly, this asymmetric expression pattern is randomized in the iv mutant and reversed in the inv mutant, indicating that LPlunc1 is downstream of iv and inv. Our results suggest a link between lipid binding/transfer and the axis development. PMID- 14745964 TI - Embryonic myogenesis pathways in muscle regeneration. AB - Embryonic myogenesis involves the staged induction of myogenic regulatory factors and positional cues that dictate cell determination, proliferation, and differentiation into adult muscle. Muscle is able to regenerate after damage, and muscle regeneration is generally thought to recapitulate myogenesis during embryogenesis. There has been considerable progress in the delineation of myogenesis pathways during embryogenesis, but it is not known whether the same signaling pathways are relevant to muscle regeneration in adults. Here, we defined the subset of embryogenesis pathways induced in muscle regeneration using a 27 time-point in vivo muscle regeneration series. The embryonic Wnt (Wnt1, 3a, 7a, 11), Shh pathway, and the BMP (BMP2, 4, 7) pathway were not induced during muscle regeneration. Moreover, antagonists of Wnt signaling, sFRP1, sFRP2, and sFRP4 (secreted frizzled-related proteins) were significantly up-regulated, suggesting active inhibition of the Wnt pathway. The pro-differentiation FGFR4 pathway was transiently expressed at day 3, commensurate with expression of MyoD, Myogenin, Myf5, and Pax7. Protein verification studies showed fibroblast growth factor receptor 4 (FGFR4) protein to be strongly expressed in differentiating myoblasts and newly formed myotubes. We present evidence that FGF6 is likely the key ligand for FGFR4 during muscle regeneration, and further suggest that FGF6 is released from necrotic myofibers where it is then sequestered by basal laminae. We also confirmed activation of Notch1 in the regenerating muscle. Finally, known MyoD coactivators (MEF2A, p/CIP, TCF12) and repressors (Twist, Id2) were strongly induced at appropriate time points. Taken together, our results suggest that embryonic positional signals (Wnt, Shh, and BMP) are not induced in postnatal muscle regeneration, whereas cell-autonomous factors (Pax7, MRFs, FGFR4) involving muscle precursor proliferation and differentiation are recapitulated by muscle regeneration. PMID- 14745965 TI - Early requirement for fgf8 function during hindbrain pattern formation in zebrafish. AB - Fibroblast growth factor (FGF) signaling is required for normal development of the vertebrate brain, including the isthmus and caudal regions of the hindbrain. Recent work in zebrafish has identified a requirement for the combination of fgf3 and fgf8 functions in specification of rhombomeres 5 and 6 (r5, r6), when evaluated at mid- and late somitogenesis stages. However, when examined earlier in development, during early somitogenesis stages, FGF8 alone is required to initiate r5 and r6 development. Both a mutation in fgf8 and injection of fgf8 targeted antisense morpholino-modified oligonucleotides result in suppression of genes normally expressed in r5 and r6 by the one- to two-somite stage. This expression recovers by the six-somite stage, and we propose that this recovery is a response to activation of fgf3 and to delayed accumulation of fgf8. These data demonstrate an early, nonredundant requirement for fgf8 function in hindbrain patterning. PMID- 14745966 TI - Ror2 knockout mouse as a model for the developmental pathology of autosomal recessive Robinow syndrome. AB - Robinow syndrome (RS) is a human dwarfism syndrome characterized by mesomelic limb shortening, vertebral and craniofacial malformations and small external genitals. We have analyzed Ror2(-/-) mice as a model for the developmental pathology of RS. Our results demonstrate that vertebral malformations in Ror2(-/ ) mice are due to reductions in the presomitic mesoderm and defects in somitogenesis. Mesomelic limb shortening in Ror2(-/-) mice is a consequence of perturbed chondrocyte differentiation. Moreover, we show that the craniofacial phenotype is caused by a midline outgrowth defect. Ror2 expression in the genital tubercle and its reduced size in Ror2(-/-) mice makes it likely that Ror2 is involved in genital development. In conclusion, our findings suggest that Ror2 is essential at multiple sites during development. The Ror2(-/-) mouse provides a suitable model that may help to explain many of the underlying developmental malformations in individuals with Robinow syndrome. PMID- 14745968 TI - Biology of insulin-like growth factors in development. AB - Insulin-like growth factors (IGFs) provide essential signals for the control of embryonic and postnatal development in vertebrate species. In mammals, IGFs act through and are regulated by a system of receptors, binding proteins, and related proteases. In each of the many tissues dependent on this family of growth factors, this system generates a complex interaction specific to the tissue concerned. Studies carried out over the last decade, mostly with transgenic and gene knockout mouse models, have demonstrated considerable variety in the cell type-specific and developmental stage-specific functions of IGF signals. Brain, muscle, bone, cartilage, pancreas, ovary, skin, and fat tissue have been identified as major in vivo targets for IGFs. Concentrating on several of these organ systems, we review here phenotypic analyses of mice with genetically modified IGF systems. Much progress has also been made in understanding the specific intracellular signaling cascades initiated by the binding of circulating IGFs to their cognate receptor. We also summarize the most relevant aspects of this research. Considerable efforts are currently focused on deciphering the functional specificities of intracellular pathways, particularly the molecular mechanisms by which cells distinguish growth-stimulating insulin-like signals from metabolic insulin signals. Finally, there is a growing body of evidence implicating IGF signaling in lifespan control, and it has recently been shown that this function has been conserved throughout evolution. Very rapid progress in this domain seems to indicate that longevity may be subject to IGF-dependent neuroendocrine regulation and that certain periods of the life cycle may be particularly important in the determination of individual lifespan. PMID- 14745969 TI - Biology of platelet-derived growth factors in development. AB - Platelet-derived growth factor (PDGF) was one of the first growth factors to be characterized, and the PDGF family of ligand and receptors has remained an archetype system for studies of the mechanisms of action of growth factors and receptor tyrosine kinases for more than two decades. The small size of the family has also facilitated genetic studies and, in particular, manipulations of the mouse PDGF and PDGF receptor genes have given important insights into the role of this family during mammalian development. These studies have shown that discrete populations of mesenchymal and neuroectodermal progenitor cells depend on PDGF signaling for their growth and distribution within developing organs. Other studies suggest that the same, or similar, cells may be targeted by exaggerated PDGF signaling in a number of pathological processes, including different types of cancer. The present review summarizes current views on the roles of PDGFs in developmental processes, and discusses the critical importance of the amount, spatial distribution, and bioavailability of the PDGF proteins for acquisition of the correct number and location of target cells. PMID- 14745970 TI - Roles of FGF receptors in mammalian development and congenital diseases. AB - Four fibroblast growth factor receptors (FGFR1-4) constitute a family of transmembrane tyrosine kinases that serve as high affinity receptors for at least 22 FGF ligands. Gene targeting in mice has yielded valuable insights into the functions of this important gene family in multiple biological processes. These include mesoderm induction and patterning; cell growth, migration, and differentiation; organ formation and maintenance; neuronal differentiation and survival; wound healing; and malignant transformation. Furthermore, discoveries that mutations in three of the four receptors result in more than a dozen human congenital diseases highlight the importance of these genes in skeletal development. In this review, we will discuss recent progress on the roles of FGF receptors in mammalian development and congenital diseases, with an emphasis on signal transduction pathways. PMID- 14745971 TI - Wnts and wing: Wnt signaling in vertebrate limb development and musculoskeletal morphogenesis. AB - In the past twenty years, secreted signaling molecules of the Wnt family have been found to play a central role in controlling embryonic development from hydra to human. In the developing vertebrate limb, Wnt signaling is required for limb bud initiation, early limb patterning (which is governed by several well characterized signaling centers), and, finally, late limb morphogenesis events. Wnt ligands are unique, in that they can activate several different receptor mediated signal transduction pathways. The most extensively studied Wnt pathway is the canonical Wnt pathway, which controls gene expression by stabilizing beta catenin in regulating a diverse array of biological processes. Recently, more attention has been given to the noncanonical Wnt pathway, which is beta-catenin independent. The noncanonical Wnt pathway signals through activating Ca(2+) flux, JNK activation, and both small and heterotrimeric G proteins, to induce changes in gene expression, cell adhesion, migration, and polarity. Abnormal Wnt signaling leads to developmental defects and human diseases affecting either tissue development or homeostasis. Further understanding of the biological function and signaling mechanism of Wnt signaling is essential for the development of novel preventive and therapeutic approaches of human diseases. This review provides a critical perspective on how Wnt signaling regulates different developmental processes. As Wnt signaling in tumor formation has been reviewed extensively elsewhere, this part is not included in the review of the clinical significance of Wnt signaling. PMID- 14745972 TI - Planar cell polarity genes and neural tube closure. AB - Closure of the neural tube is essential for normal development of the brain and spinal cord. Failure of closure results in neural tube defects (NTDs), common and clinically severe congenital malformations whose molecular mechanisms remain poorly understood. On the other hand, it is increasingly well established that common molecular mechanisms are employed to regulate morphogenesis of multicellular organisms. For example, signaling triggered by polypeptide growth factors is highly conserved among species and utilized in multiple developmental processes. Recent studies have revealed that the Drosophila planar cell polarity (PCP) pathway, which directs position and direction of wing hairs on the surface of the fly wing, is well conserved, and orthologs of several genes encoding components of the pathway are also found in vertebrates. Interestingly, in vertebrates, this signaling pathway appears to be co-opted to regulate "convergent extension" cell movements during gastrulation. Disruption of vertebrate PCP genes in Xenopus laevis or zebrafish causes severe gastrulation defects or the shortening of the trunk, as well as mediolateral expansion of somites. In Xenopus, in which the neural tube closes by elevation and fusion of neural folds, inhibition of convergent extension can also prevent neural tube closure causing a "spina bifida-like" appearance. Furthermore, several of the genes involved in the PCP pathway have recently been shown to be required for neural tube closure in the mouse, since mutation of these genes causes NTDs. Therefore, understanding the mechanisms underlying the establishment of cell polarity in Drosophila may provide important clues to the molecular basis of NTDs. PMID- 14745973 TI - Nodal signaling and vertebrate germ layer formation. AB - The understanding of germ layer formation in vertebrates began with classical experimental embryology. Early in the 20th century, Spemann and Mangold (1924) identified a region of the early embryo capable of inducing an entire embryonic axis. Termed the dorsal organizer, the tissue and the activity have been shown to exist in all vertebrates examined. In mice, for example, the activity resides in a region of the gastrula embryo known as the node. Experiments by the Dutch embryologist Nieuwkoop (1967a, 1967b, 1973, 1977) showed that a signal derived from the vegetal half of the amphibian embryo is responsible for the formation of mesoderm. Nieuwkoop's results allowed the development of in vitro assays that led, in the late 1980s and early 1990s, to the identification of growth factors essential for germ layer formation. Through more recent genetic investigations in mice and zebrafish, we now know that one class of secreted growth factor, called Nodal because of its localized expression in the mouse node, is essential for formation of mesoderm and endoderm and for the morphological rearrangements that occur during gastrulation. PMID- 14745974 TI - TGF-beta signaling in human skeletal and patterning disorders. AB - Members of the transforming growth factor beta (TGF-beta) family of multifunctional peptides are involved in almost every aspect of development. Model systems, ranging from genetically tractable invertebrates to genetically engineered mice, have been used to determine the mechanisms of TGF-beta signaling in normal development and in pathological situations. Furthermore, mutations in genes for the ligands, receptors, extracellular modulators, and intracellular signaling molecules have been associated with several human disorders. The most common are those associated with the development and maintenance of the skeletal system and axial patterning. This review focuses on the mechanisms of TGF-beta signaling with special emphasis on the molecules involved in human disorders of patterning and skeletal development. PMID- 14745975 TI - PTHrP, PTH, and the PTH/PTHrP receptor in endochondral bone development. AB - Endochondral bone development is a fascinating story of proliferation, maturation, and death. An understanding of this process at the molecular level is emerging. In particular, significant advances have been made in understanding the role of parathyroid-hormone-related peptide (PTHrP), parathyroid hormone (PTH), and the PTH/PTHrP receptor in endochondral bone development. Mutations of the PTH/PTHrP receptor have been identified in Jansen metaphyseal chondrodysplasia, Blomstrand's lethal chondrodysplasia, and enchondromatosis. Furthermore, genetic manipulations of the PTHrP, PTH, and the PTH/PTHrP receptor genes, respectively, have demonstrated the critical role of these proteins in regulating both the switch between proliferation and differentiation of chondrocytes, and their replacement by bone cells. A future area of investigation will be the identification of downstream effectors of PTH, PTHrP, and PTH/PTHrP receptor activities. Furthermore, it will be of critical importance to study how these proteins cooperate and integrate with other molecules that are essential for growth plate development. PMID- 14745976 TI - From the cradle to the clinic: VEGF in developmental, physiological, and pathological angiogenesis. AB - Formation of new blood vessels, which is fundamental in embryonic development, occurs through a combination of angiogenesis and vasculogenesis. Angiogenesis also plays a vital role postnatally, especially in reparative processes such as wound and fracture healing. Some of these events, especially in fracture healing, recapitulate processes observed in developmental angiogenesis. However, dysregulated angiogenesis is well documented to underlie a number of pathological disorders, including rheumatoid arthritis (RA). The vascular endothelial growth factor (VEGF)/VEGF receptor system is the best characterized regulator of angiogenesis. VEGF is expressed in a range of cells in response to soluble mediators (such as cytokines and growth factors), cell-bound stimuli (such as CD40 ligand), and environmental factors (such as hypoxia). As a consequence, this molecule is vital in the modulation of physiological and pathological angiogenesis. This review will focus in particular on the role played by VEGF in embryogenesis and skeletal growth, in fracture healing (in which increased angiogenesis is likely to be beneficial in promoting union), and in RA (in which excessive angiogenesis is thought to play a significant role in disease pathogenesis). In the not-too-distant future, targeting VEGF may prove to be of benefit in the treatment of diseases associated with excessive or aberrant angiogenesis, such as malignancies and RA. PMID- 14745978 TI - Does terbutaline damage the developing heart? AB - BACKGROUND: Beta(2)-Adrenoceptor (betaAR) agonists, such as terbutaline, are widely used to arrest preterm labor. They also cross the placenta where they stimulate receptors in fetal tissues, which in turn use betaAR input for trophic control of cell replication and differentiation. METHODS: As rats are altricial, we administered terbutaline in two different postnatal exposure periods (10 mg/kg given daily on Days 2-5 or 11-14). RESULTS: Hearts were examined twenty-four hours after the last dose and on postnatal day 30 for cardiac damage. Neither treatment paradigm caused an increase in cardiac abnormalities compared to controls but quantitative analysis of the number of nuclei indicated reductions in females. CONCLUSIONS: These findings do not support earlier case reports of outright myocardial necrosis after terbutaline tocolysis in human infants. Nevertheless, the significant statistical association between terbutaline and cardiac anomalies in epidemiological studies suggest that terbutaline may sensitize the developing heart to other insults that affect development. PMID- 14745979 TI - Mercury, cadmium, and arsenite enhance heat shock protein synthesis in chick embryos prior to embryotoxicity. AB - BACKGROUND: Cells respond to adverse environmental stimuli by enhancing the expression of specific genes, the products of which include a suite of proteins known as heat shock proteins (hsps), a response often attributed to cellular protection. METHODS: In this study, we characterized alterations in hsp expression in chick embryos (Hamburger-Hamilton stage 17, 72 h) exposed in ovo to arsenite (As), mercury (Hg), and cadmium (Cd), known developmental toxicants. Embryos were incubated for 2 h following exposure to 3, 10, 30, or 100 nmol metal, or for 2, 4, 12, or 24 h following treatment with 10 nmol metal. RESULTS: An enhanced de novo synthesis of 24-, 70-, and 90-kD, 70- and 90-kD, and 70-kD proteins was observed with As, Hg, and Cd treatments, respectively. These responses were transient; apparent rates of protein synthesis were maximal 2-4 h after exposure and returned to control rates by 24 h. Actinomycin D experiments demonstrated that arsenite-induced expression of these proteins is transcriptionally regulated. Immunoblotting experiments identified the 24-, 70-, and 90-kD proteins as the heat shock proteins hsp24, hsp70, and hsp90, respectively. Exposure duration-related abnormalities were noted in the neural tube with all metals and in the ganglia and somites with Cd and As. Retina, allantois, and limb defects were specific to Cd-treated embryos, and branchial arch defects were specific to As-treated embryos. CONCLUSIONS: The data support metal-induced developmental abnormalities, which are preceded by synthesis of stress proteins. PMID- 14745980 TI - Prenatal window of susceptibility to perfluorooctane sulfonate-induced neonatal mortality in the Sprague-Dawley rat. AB - The critical period for increased neonatal mortality induced by perfluorooctane sulfonate (PFOS) exposure was evaluated in the rat. Timed-pregnant Sprague-Dawley rats were treated by oral gavage with 25 mg/kg/d PFOS/K(+) on four consecutive days (gestation days (GD) 2-5, 6-9, 10-13, 14-17, or 17-20) or with 0, 25, or 50 mg/kg/d PFOS/K(+) on GD 19-20. Controls received vehicle (10 ml/kg 0.5% Tween-20) on these days. Maternal weight gain was reduced in treated animals during dosing, as were food and water consumption. Following a 4-day treatment, litter size at birth was unaffected while pup weight was similarly reduced in the three earliest PFOS groups. All PFOS groups experienced decreases in survival while controls remained near 100%. Neonatal survival decreased in groups dosed later during gestation, approaching 100% with dosing on GD 17-20. Most deaths occurred before postnatal day (PND) 4, with the majority in the first 24 hours. Maternal serum PFOS levels on GD 21 were higher in groups exhibiting higher mortality. Following a 2-day treatment, PFOS groups experienced significant pup mortality by PND 1. Neonatal mortality continued through PND 5, when survival was 98, 66, and 3% for the 0, 25, and 50 mg/kg groups, respectively. Pup weight was reduced in treated groups with surviving litters. Gross dissection and histological examination of lungs revealed differences in maturation between control and treated animals on PND 0. We conclude that exposure to PFOS late in gestation is sufficient to induce 100% pup mortality and that inhibition of lung maturation may be involved. PMID- 14745981 TI - Induction of developmental toxicity in mice treated with Alstonia scholaris (Sapthaparna) In utero. AB - BACKGROUND: The teratogenic effect of hydroalcoholic extract of Alstonia scholaris (ASE) was studied in the pregnant Swiss albino mice administered with 0, 60, 120, 240, 360, and 480 mg/kg ASE on Day 11 of gestation. METHODS: Females were allowed to complete the term and parturiate. The litters were monitored regularly for mortality, growth retardation, congenital malformations, and appearance of physiological markers up to 7 weeks post-parturition (p.p.). RESULTS: The administration of 60, 120, 180, and 240 mg/kg ASE to the pregnant mice on Day 11 did not induce mortality, congenital malformations, or alter the normal growth patterns. A further increase in the herbal extract dose up to 360 or 480 mg/kg resulted in a dose dependent increase in the mortality, growth retardation, and congenital malformations, characterized mainly by bent tails and syndactyly. The administration of higher doses (360 or 480 mg) of ASE also caused a significant delay in the morphological parameters such as fur development, eye opening, pinna detachment, and vaginal opening. The incisor eruption and testes decent were found to be delayed in litters born to the mothers treated with 240 480 mg/kg ASE. CONCLUSIONS: Our study indicates clearly that ASE treatment caused teratogenic effect only at doses above 240 mg/kg (>20% of LD(50)). Lower doses had no developmental toxicity. PMID- 14745982 TI - Role of thyroid hormones in human and laboratory animal reproductive health. AB - The highly conserved nature of the thyroid gland and the thyroid system among mammalian species suggests it is critical to species survival. Studies show the thyroid system plays a critical role in the development of several organ systems, including the reproductive tract. Despite its highly conserved nature, the thyroid system can have widely different effects on reproduction and reproductive tract development in different species. The present review focuses on assessing the role of thyroid hormones in human reproduction and reproductive tract development and comparing it to the role of thyroid hormones in laboratory animal reproduction and reproductive tract development. The review also assesses the effects of thyroid dysfunction on reproductive tract development and function in humans and laboratory animals. Consideration of such information is important in designing, conducting, and interpreting studies to assess the potential effects of thyroid toxicants on reproduction and development. PMID- 14745983 TI - What do we know about the reproductive and developmental risks of herbal and alternate remedies? PMID- 14745984 TI - Interactions of herbal products with conventional medicines and potential impact on pregnancy. PMID- 14745985 TI - Potential toxicities of herbal therapies in the developing fetus. PMID- 14745986 TI - Herbal medicine use in pregnancy: a new frontier in clinical teratology. PMID- 14745987 TI - Pregnancy outcomes in women using herbal therapies. PMID- 14745988 TI - Regulatory issues concerning the safety, efficacy and quality of herbal remedies. AB - Herbal remedies and alternative medicines are used throughout the world, and in the past herbs were often the original sources of most drugs. Today we are witnessing an increase in herbal remedy use throughout the Western world raising the question as to how safe are these preparations for the unborn fetus? Many women use herbal products during pregnancy. The dilemma facing most regulatory authorities is that the public considers these products as either traditional medicines or natural food supplements. The user sees no reason for regulation. Most countries have laws concerning foods, drugs, and cosmetics, the details of which seldom clearly define to what section of the law and regulations alternative remedies belong. In most countries alternative remedies are regulated as foods, provided that no medicinal claim is made on the label. The global regulatory sector, however, is changing rapidly. The Therapeutic Goods Administration (TGA) in Australia created a Complimentary Medicines Evaluation Committee in late 1997 to address this issue, and Canada has created a new Natural Health Products Directorate in the realigned Therapeutic Products and Foods Branch in 2000. In parallel, the European Agency for the Evaluation of Medicinal Products has drafted test procedures and acceptance criteria for herbal drug preparations and herbal medicinal products. In the US, the Food and Drug Administration classifies these natural products as dietary supplements. Manufacturers must label a dietary supplement thus: "this statement has not been evaluated by the FDA [, and] this product is not intended to diagnose, treat, cure or prevent any disease." Whether these products are foods or drugs is undecided. To add complexity to this issue, most of the potential deleterious effects of natural products on the unborn may be related to hormonal effects (e.g., phytoestrogens) and nutriceutical drug interactions (e.g., St. John's Wort and antidepressants), rather than direct embryotoxicity per se. We suggest that ensuring quality of herbal products should receive immediate attention by regulatory authorities, before embarking on the more arduous tasks of safety and efficacy. PMID- 14745989 TI - Species comparison of postnatal CNS development: functional measures. PMID- 14745990 TI - Evaluation of testicular toxicology: a synopsis and discussion of the recommendations proposed by the Society of Toxicologic Pathology. AB - BACKGROUND: Detection of chemically induced effects on male fertility and on testicular spermatogenesis in particular, has become of increasing concern. More stringent regulatory guidelines, introduced by ICH, EPA and OECD (Table 1) have raised the awareness of toxicologists and pathologists for the need to conduct sensitive and careful evaluation of the male reproductive tract for potential toxic effects of administered compounds. With it has come confusion and in many cases, inappropriate procedures, often based on misunderstanding of what is required and on inadequate understanding of spermatogenesis. This article summarizes and discusses the main recommendations recently proposed by the Society of Toxicologic Pathology on recommended approaches for the evaluation of testicular and epididymal toxicity [Lanning LL, Creasy DM, Chapin RE, Mann PC, Barlow NJ, Regan KS, Goodman DG. Toxicologic Pathology 30:518-531, 2002]. The major recommendations are: Use sexually mature animals to evaluate effects on spermatogenesis. Sample left and right testes and epididymides and record organ weights. Use modified Davidson's fixative to fix testes from all species from studies of 13 wks duration and less. Examine transverse sections of the testes (including part of the rete), and longitudinal sections of the epididymides. Embed tissues in paraffin wax. For rodent studies up to 28 days, examine periodic acid-Schiff's-hematoxylin stained sections. For all other studies examine hematoxylin and eosin stained sections. Microscopic evaluation of the testis should be a qualitative evaluation carried out with an awareness of the spermatogenic cycle. Quantitative procedures are inappropriate for screening studies. Nomenclature and grading of findings for spermatogenic disturbances will vary on a case by case basis. PMID- 14745991 TI - Preliminary screening study of reproductive outcomes after exposure to yarrow in the pregnant rat. AB - BACKGROUND: Yarrow (Achillea millefolium) is used for the treatment of diarrhea, as a urinary antiseptic, hypotensive, and diuretic. Yarrow also has traditionally been used as an abortifacient, emmenagogue, contraceptive, and for stimulating uterine contractions. For this reason, it is contra-indicated for use in pregnancy. There has, however, been little scientific research carried out to either confirm or refute this recommendation. METHODS: Female rats were dosed, orally by gavage using 56 times the human dose of yarrow daily on either gestation days (GD) 1-8 or GD 8-15. Two groups of controls were included; the first received water and the second received an equivalent dose of ethanol to that found in the yarrow preparation over the two gestation periods. On GD 20, rats were sacrificed, placentae were weighed, and corpora lutea counted. The fetuses were weighed and examined for signs of external, internal or skeletal malformations. RESULTS: The dose used was not materno-toxic. There was no increase in pre- or post-implantation losses suggesting that yarrow was neither an abortifacient nor a contraceptive. Placental weights were increased in rats treated with yarrow on GD 8-15 compared to water and ethanol controls and on GD 1 8 compared to water control fetuses. Body weight was reduced in fetuses exposed to yarrow on GD 8-15 compared to water control fetuses. There was no difference in incidence of external or internal malformations. CONCLUSIONS: In the present study it was found that yarrow, when administered to rats at 56 times the human dose, was associated with reduced fetal weight and increased placental weight. In the absence of a no observable effect level for these variables it must be concluded that the consumption of yarrow is contraindicated during pregnancy until further investigations have been carried out. PMID- 14745992 TI - Embryo/fetal development studies with hydroxypropyl methylcellulose acetate succinate (HPMCAS) in rats and rabbits. AB - BACKGROUND: Hoshi et al. [Hoshi et al. J Toxicol Sci 10(Suppl):187-255, 1985a,b,c,d] evaluated the potential for hydroxypropyl methylcellulose acetate succinate (HPMCAS) to produce developmental and reproductive toxicity in a series of studies that included rat and rabbit teratology studies, a rat fertility study, and a rat peri- and postnatal study. The authors concluded that there were no compound-related findings. In the cesarean-section phase of the rat teratology study, however, clubfoot was reported for 0.8, 2.1, 5.5, and 4.1% of fetuses in the control, 625, 1250, and 2500 mg/kg groups, respectively. There were no significant increases in external anomalies, but the apparent dose-related increase in clubfoot was not specifically addressed. In the rabbit teratology study, the number of litters evaluated (12-13 per group) was not consistent with current regulatory guidelines. Therefore, to definitively establish the potential of HPMCAS to produce developmental toxicity, embryo/fetal development studies were carried out in rats and rabbits. METHODS: Groups of 20 pregnant Sprague Dawley rats and New Zealand White rabbits were dosed with 0, 50, 150, 625, or 2500 mg/kg HPMCAS from gestational day (GD) 6-17 or GD 7-19 for rats and rabbits, respectively. Fetuses were collected by cesarean section and examined for external, visceral and skeletal development. RESULTS: No developmental toxicity was observed as a result of HPMCAS exposure demonstrating that maternal HPMCAS exposure during gestation does not induce developmental anomalies. There were no findings of clubfoot or other limb anomalies in these studies at dose levels equivalent to those that were previously associated with a possible increase in clubfoot. CONCLUSIONS: The conclusion of the earlier study indicating that treatment with HPMCAS at doses up to and including 2500 mg/kg did not produce developmental toxicity was confirmed with these studies. It is likely that the clubfoot noted in the earlier rat teratology study was a misdiagnosis or artifact. PMID- 14745994 TI - The Dutch-Belted rabbit: an alternative breed for developmental toxicity testing. AB - BACKGROUND: Historical control data play a crucial role in the interpretation of observations made from developmental and reproductive toxicity studies. METHODS: In our effort to develop the Dutch-Belted rabbit as an alternative breed of rabbit for testing of small molecule pharmaceuticals, data collected from 236 untreated pregnant Dutch Belted rabbits are summarized and compared to data collected from 350 untreated pregnant New Zealand White Rabbits. RESULTS: The data presented include mean maternal body weight and food consumption values, mean maternal hematological and serum biochemical values collected during gestation, maternal reproductive (laparotomy) parameters collected at cesarean section, and incidences of spontaneous fetal morphological alterations collected during 1999-2002. Due to their smaller size the use of this breed of rabbit would require approximately 40% less drug than studies conducted with New Zealand White rabbits. Because the Dutch-Belted rabbit is pigmented this breed also offers the ability to test the potential effects of various xenobiotics on melanocyte development and to evaluate the potential sequela of drugs that have a propensity to bind melanin in the skin and pigmented retinal epithelium. CONCLUSIONS: Based on these data, the Dutch-Belted rabbit is considered to be a practical alternative to use of the New Zealand White rabbit for these types of studies. PMID- 14745993 TI - Experimental studies on reproductive toxicologic effects of lamotrigine in mice. AB - BACKGROUND: Virtually all antiepileptic drugs (AED) tested so far have been found to be teratogenic. The second generation AED possess a number of therapeutic advantages over the older ones. There are, however, very little data on their effects on embryonic development. A recent report suggests that lamotrigine (LTG) can be teratogenic to human fetuses. With only a few cases of prenatal exposure to LTG in the record, however, it has not been possible to establish a recognizable pattern of malformations in the infants of LTG-treated mothers. OBJECTIVES: The objectives of the present study were to evaluate the reproductive toxic effects of LTG . RESULTS: Single (50-200 mg/kg) or multiple doses (25, 50, 75 mg/kg) of LTG were administered by intraperitoneal (i.p.) injection (note that the therapeutic administration is oral) to groups of TO mice on gestation day (GD) 7 or 8. Fetuses were collected on GD 18. Maternal toxic effects including a dose-related mortality, a high incidence of abortion, embryo lethality, congenital malformations and intrauterine growth retardation (IUGR) were observed in the LTG-treated group. Administration of LTG in multiple low doses resulted in a better maternal survival and increased incidence of embryonic resorption and malformations with increasing dose; IUGR was significant but not dose-dependent. The malformations characteristic of the LTG multiple low dose group fetuses included maxillary-mandibular hypoplasia, exencephaly, cleft palate, median facial cleft, urogenital anomalies and varying degrees of caudal regression. Skeletal malformations and developmental delay of the skeleton were observed both in single and multiple dose groups. CONCLUSIONS: The results of this study indicate that LTG administered i.p. at high doses can induce intrauterine growth retardation and at low multiple doses causes a dose-dependent increase in embryonic resorption, craniofacial and caudal malformations as well as maternal toxicity in the mouse. Previous studies in other laboratories have used oral route of exposure and concluded that there are no teratogenic effects of LTG at dose levels that are not maternally toxic. PMID- 14746002 TI - [Lead poisoning in children:an old problem requiring new solutions]. PMID- 14746004 TI - [Trends in atmospheric concentrations of lead in the metropolitan area of Mexico city, 1988-1998]. AB - OBJECTIVE: To describe the trends in airborne lead levels and their association with the control measures implemented from 1988 to 1998 to phase out lead from gasoline that included the introduction of a lead-free gasoline and the use of catalytic converters in automobiles in Mexico City. MATERIAL AND METHODS: Data of atmospheric lead levels were obtained from the 9 field stations included in the local air quality monitoring network (RMMA). Trends for airborne lead are shown by monitor, monitoring area and for the entire metropolitan zone. Analysis of data was performed using linear regression models including auto correlation analysis. The models were validated by standard diagnostic techniques using student residuals analysis. RESULTS: The introduction of lead-free gasoline and the use of catalytic converters decreased the airborne lead levels by 23% while the slow elimination of lead from gasoline produced a cumulative decrease of 89% during the period and in all areas studied. CONCLUSIONS: The strategy implemented to reduce sources of airborne lead pollution had an important impact on atmospheric lead levels. Health risks of this environmental hazard are now primarily due to less important sources and to particular occupational exposures. The English version of this paper is available at: http://www.insp.mx/salud/index.html. PMID- 14746003 TI - Determining lead sources in Mexico using the lead isotope ratio. AB - OBJECTIVE: Lead poisoning can, in some cases, be traced to a specific route or source of exposure on the basis of the individual's blood lead isotope ratio. To assess the major source of lead exposure among women residing in Mexico City, we compared blood, ceramic, and gasoline lead isotope ratios. MATERIAL AND METHODS: The study population, randomly selected from participants of a large trial, (1/1996-12/1996) comprised of 16 women whose lead levels exceeded 10 micrograms/dl and who reported using lead-glazed ceramics. Lead isotope ratios were performed on a Perkin Elmer 5000 Inductively Coupled Plasma Mass Spectrometer (ICP-MS) interfaced with a Perkin Elmer HGA-600MS Electrothermal Vaporization System (ETV). RESULTS: The isotope ratios (206Pb/204Pb, 207Pb/204Pb, and 208Pb/204Pb) of both the blood specimens and their corresponding ceramic specimens were highly correlated, with r = 0.9979, r2 = 0.9958, r = 0.9957, r2 = 0.9915 and r = 0.9945, r2 = 0.9890 values for the three isotope ratios, respectively, suggesting that the lead exposure most likely resulted from the use of these ceramic. Measurements of lead isotope ratios from leaded gasoline in use at the time of blood sampling, differed from those in blood and ceramics. CONCLUSIONS: Determining lead isotope ratios can be an efficient tool to identify a major source of lead exposure and to support the implementation of public health prevention and control measures. This paper is available too at: http://www.insp.mx/salud/index.html. PMID- 14746005 TI - [Blood lead levels and exposure factors in children of Morelos state, Mexico]. AB - OBJECTIVE: To assess blood lead levels and lead exposure factors in children living in Morelos State, Mexico. MATERIAL AND METHODS: A cross-sectional study was conducted between June and October 1996, in 232 children aged 1-12 years, at Hospital del Nino Morelense de Cuernavaca, Morelos, Mexico. Blood lead levels were measured by anodic voltameter, and exposure factors were collected by questionnaire. The lead concentration value was log transformed for statistical analysis. Odds ratios were obtained for some risk factors. The statistical significative risk factors were later analyzed with ANOVA. RESULTS: A total of 232 children were recruited (50% female); 73% resided in Cuernavaca City. The geometric mean blood lead level was 6.7 micrograms/dl; 29.7% of the children had levels over 10 micrograms/dl; 66% reported use of lead glazed pottery for cooking, 36% for storing food, and 19% for drinking. CONCLUSIONS: Blood lead levels were similar to those reported in other Mexican children studies, after the reduction of lead in gasoline. The main risk factors were use of lead glazed pottery and vehicle traffic intensity near the household. These results will be useful for future prevention and control interventions. This paper is available too at: http://www.insp.mx/salud/index.html. PMID- 14746006 TI - Determinants of blood-lead levels in children in Callao and Lima metropolitan area. AB - OBJECTIVES: To determine blood lead levels in urban populations of children (n = 2,510) and women (n = 874) in the early postpartum in certain districts of Lima and Callao, and to correlate those levels with particular exposures. MATERIAL AND METHODS: Between July 1998 and January 1999 cross sectional study was conducted. The study population was selected using three sampling strategies in the government operated school system and from public pediatric and maternity hospitals at Lima and Callao, Peru. Study personnel were trained to collect finger stick blood samples with a protocol that minimizes external lead contamination. Lead determinations in blood and environmental samples were performed at the study site using portable anodic striping voltamenters. To determine the simultaneous effects of different predictors on blood lead levels, multivariate regression models were used to estimate adjusted mean differences. RESULTS: The mean blood lead level in the children studied was 9.9 micrograms/dl ranging from 1 microgram/dl to 64 micrograms/dl with 29% of the children displaying values greater than 10 micrograms/dl and 9.4% at levels greater than 20 micrograms/dl. Among the women, the mean was 3.5 micrograms/dl (SD = 2.4 micrograms/dl), and 2.4% (n = 21) displayed levels greater than 10 micrograms/dl. Important differences were observed between the sample locations, and the highest levels were documented in the port region near Callao. The mean level of blood lead in this group was 25.6 micrograms/dl (SD = 4.6 micrograms/dl), while among the rest of the sample it was 7.1 micrograms/dl (SD = 5.1 micrograms/dl). The presence of a mineral storage area signified a difference in exposure in excess of 13 micrograms/dl for children living near the port area in contrast to the other children who were not as close to such fixed sources of lead exposure. For the participants in Lima, the risk of showing levels above 10 micrograms/dl was associated with exposure to high vehicular traffic. CONCLUSIONS: In metropolitan Lima, we conclude that the mean blood lead levels of the populations studied were not alarming and that a positive health impact can be made by a reduction of lead in gasoline. With regard to the port area, the study demonstrates that the presence of mineral storage areas pose a detrimental risk factor for the health of the children living in this area. The English version of this paper is available too at: http://www.insp.mx/salud/index.html. PMID- 14746007 TI - [Effects of lead exposure on children's health]. AB - Lead is an element that has no known physiologic function in humans but adversely affects a variety of fundamental biochemical processes. A large body of evidence shows adverse health effects of lead in children at levels common in populations around the world. Acute lead poisoning with encephalopathy, though infrequent, is life-threatening, requiring timely and aggressive treatment. It should be considered in the differential diagnosis of any unexplained illness that includes anemia, seizures, lethargy, abdominal pain, or recurrent vomiting. Many more children are harmed by chronic, subclinical effects of low-level lead exposure that include impaired cognitive development, adverse effects on behavior, subtle impairment of hearing acuity, and reduced height. Available evidence indicates that the only effective interventions to prevent low level lead toxicity are those that control lead exposure. The English version of this paper is available at: http://www.insp.mx/salud/index.html. PMID- 14746008 TI - [Managing childhood lead poisoning]. AB - This paper reviews the clinical management of children with lead poisoning. A first step is to define the measures to be used in their assessment and be aware of the limitations. Measurements of blood lead levels can be made on anticoagulated whole blood samples using either: atomic absorption spectroscopy or anodic stripping voltametry. However a more accurate method is fluorescent RX'ray of the skeleton or systematic biochemical tests of lead levels in urine. Remedies include elimination of lead in the environment, changes in children's behavior and dietary checks for adequate calcium and iron intake. Chelation therapy, using Ca edetate and succimer eliminates lead from the skeleton, which is then quickly excleted using a cathartic to help prevent re-absorption. Chelation may save lives where BLLs are very high. There is usually a short term reduction of BLLs with a subsequent rise. Serious cases may require repeat therapies. Chelation should be considered in children with BLLs > = 45 micrograms/dl. Chelation therapy reduces BLLs and associated symptoms. However cognitive decline may be irreversible, indicating that emphasis should be on prevention rather than cure. The English version of this paper is available at: http://www.insp.mx/salud/index.html. PMID- 14746009 TI - [Environmental intervention in sites contaminated by lead: the United States of America experience]. AB - For a decade, state and federal agencies have worked jointly with communities throughout the USA, with the objective of measuring the health-risk from lead pollution in residential zones. Often these communities have been linked with facilities previously associated with activities like mining and metallurgy; nevertheless, there are other industries like paint manufacturing and battery recycling, that have also been identified as lead pollution sources. The vast experience in cleaning up the contaminated sites has shown that ample programs designed to identify and handle the exposure routes can help, in an effective manner, to diminish blood lead levels (BLL) in susceptible populations, such as in young children. Environmental intervention programs are more effective when the affected communities carry out health education/intervention programs, geared towards the development of individualized strategies for handling the risk implied by the presence of lead in the atmosphere. The English version of this paper is available at: http://www.insp.mx/salud/index.html. PMID- 14746010 TI - [Establishing lead in air and water standards in the United States of America]. AB - Lead regulations and standards have resulted in a lower exposure to lead in the general population of the United States of America. This paper highlights some of the know-how developed through lead-containing experiences, particularly regarding lead content in air and water. The availability of a solid and clear scientific knowledge is central to the success of these policies. Collateral effects of some air-related standards show the extent to which problem-oriented efforts may be beneficial in other areas, inasmuch as they may have untoward consequences if careful planning and evaluation are not considered. Finally, this paper presents a discussion of the differences between centralized and decentralized approaches to lead exposure control. The English version of this paper is available at: http://www.insp.mx/salud/index.html. PMID- 14746011 TI - [Use of blood lead data to evaluate and prevent childhood lead poisoning in Latin America]. AB - Exposure to lead is a widespread and serious threat to the health of children in Latin America. Health officials should monitor sources of exposure and health outcomes to design, implement, and evaluate prevention and control activities. To evaluate the magnitude of lead as a public health problem, three key elements must be defined: I) the potential sources of exposure, 2) the indicators to evaluate health effects and environmental exposure, and 3) the sampling methods for the population at risk. Several strategies can be used to select the study population depending on the study objectives, the time limitations, and the available resources. If the objective is to evaluate the magnitude and sources of the problem, the following sampling methods can be used: I) population-based random sampling; 2) facility-based random sampling within hospitals, daycare centers, or schools; 3) target sampling of high risk groups; 4) convenience sampling of volunteers; and 5) case reporting (which can lead to the identification of populations at risk and sources of exposures). For all sampling methods, information gathering should include the use of a questionnaire to collect general information on the participants and on potential local sources of exposure, as well as the collection of biological samples. In interpreting data, one should consider the type of sampling used and the non-response rates, as well as factors that might influence blood lead measurements, such as age and seasonal variability. Blood lead measurements should be integrated in an overall strategy to prevent lead toxicity in children. The English version of this paper is available at: http://www.insp.mx/salud/index.html. PMID- 14746012 TI - [Childhood health and lead in Argentina]. PMID- 14746013 TI - Human health risk reduction due to lead exposure in Brazil. PMID- 14746014 TI - [Pattern of decrease of blood lead levels in the population of Costa Rica]. PMID- 14746015 TI - [Study of levels of lead in water for human consume in Cuba]. PMID- 14746016 TI - [Prevalence of lead in blood in scholar children in Santiago of Chile]. PMID- 14746017 TI - [Lead contamination in the suburb of La Teja; Montevideo, Uruguay]. PMID- 14746018 TI - [Levels of lead in Venezuelan people with no occupational exposure. ]. PMID- 14746019 TI - [Epidemiologic research on lead levels in children and indoor air in Ciudad Juarez , Chihuahua, related with a smelter in El Paso, Texas. 1975]. PMID- 14746020 TI - Cervical cancer: advances in prevention and knowledge of its etiology. PMID- 14746021 TI - Cervical cancer prevention: the case of developing countries. PMID- 14746022 TI - Cervical cancer: new perspectives for diagnosis, prevention and control in developing countries. PMID- 14746023 TI - Incidence and mortality of cervical cancer in Latin America. AB - Cervical cancer incidence and mortality estimates for 2000 are presented for the 21 Latin American countries, using estimates from the statistical package GLOBOCAN 2000. Additional data on time-trends are also presented, using the WHO mortality database. By the year 2000, some 76,000 cervical cancer and almost 30,000 deaths were estimated for the whole region, which represent 16% and 13% of the world burden, respectively. Thus, Latin American countries are among those with highest incidence rates in the world, together with countries from Sub Saharan Africa, South and South East Asia. Variation in incidence among countries is large. Very high rates are found in Haiti (ASR 93.9 per 100,000), Nicaragua (ASR 61.1 per 100,000) and Bolivia (ASR 58.1 per 100,000). It seems unlikely that differences in risks in the region can be explained as the result of screening activities. Several descriptive studies carried out to evaluate the screening programmes in Latin America have pointed out problems related to insufficient coverage and frequency of screening. Other related problems include inadequate collection and reading of cytological samplings as well as incomplete follow-up of women after the test. The main challenge for Latin America countries remains on how to organize effective screening programmes, and for this, a real and urgent commitment from public health services and decision-makers in the region is needed. This paper is available too at: http://www.insp.mx/salud/index.html. PMID- 14746024 TI - Cervical cancer, a disease of poverty: mortality differences between urban and rural areas in Mexico. AB - OBJECTIVE: To examine cervical cancer mortality rates in Mexican urban and rural communities, and their association with poverty-related factors, during 1990 2000. MATERIAL AND METHODS: We analyzed data from national databases to obtain mortality trends and regional variations using a Poisson regression model based on location (urban-rural). RESULTS: During 1990-2000 a total of 48,761 cervical cancer (CC) deaths were reported in Mexico (1990 = 4,280 deaths/year; 2000 = 4,620 deaths/year). On average, 12 women died every 24 hours, with 0.76% yearly annual growth in CC deaths. Women living in rural areas had 3.07 higher CC mortality risks compared to women with urban residence. Comparison of state CC mortality rates (reference = Mexico City) found higher risk in states with lower socio-economic development (Chiapas, relative risk [RR] = 10.99; Nayarit, RR = 10.5). Predominantly rural states had higher CC mortality rates compared to Mexico City (lowest rural population). CONCLUSIONS: CC mortality is associated with poverty-related factors, including lack of formal education, unemployment, low socio-economic level, rural residence and insufficient access to healthcare. This indicates the need for eradication of regional differences in cancer detection. This paper is available too at: http://www.insp.mx/salud/index.html. PMID- 14746025 TI - Epidemiology of human papillomavirus infections: new options for cervical cancer prevention. AB - In the last two decades, the cervical cancer puzzle has become a coherent description that includes the identification of human papillomavirus (HPV) as the sexually transmitted etiologic agent and the characterization of the major determinants of HPV acquisition. Triage studies have consistently shown that HPV testing is more sensitive that repeated cytology in identifying underlying high grade lesions in women with atypical squamous cells of undetermined significance (ASCUS). Studies that reflect primary screening conditions have shown that the sensitivity of HPV tests is higher than standard cytology in detecting high-grade lesions whereas the specificity is similar only in women aged 30-35 and above. HPV vaccines have an intrinsic attraction as a preventive strategy in populations with limited resources. However, vaccines designed to widespread use are still in development and testing phases. Time is ripe for exploring in depth the clinical implications of current achievements and to devise novel strategies for the prevention of cervical cancer. This paper is available too at: http://www.insp.mx/salud/index.html. PMID- 14746026 TI - The 2001 Bethesda system. AB - The Bethesda system is a system of terminology for reporting the results of cervical cytology. It was developed in 1988, and is now widely used in the United States. This system was updated in April 2001. The most important modifications are the following: a) elimination of the category "satisfactory but limited by", b) the reintegration of benign modifications in the normal category, c) "atypical squamous cells of undetermined significance" is now named "atypical squamous cells" and subdivided into "atypical squamous cells of undetermined significance" and "cannot exclude high grade squamous intraepithelial lesion"; and d) endocervical adenocarcinoma in situ which is now a separate entity. Low grade squamous intraepithelial lesions and high grade squamous intraepithelial cells remain unchanged. Recommendations have been proposed concerning automated review and ancillary testing. This paper is available too at: http://www.insp.mx/salud/index.html. PMID- 14746027 TI - The male role in cervical cancer. AB - Experimental, clinical, and epidemiological evidence strongly suggests that genital Human Papillomaviruses (HPVs) are predominantly sexually transmitted. Epidemiological studies in virginal and HPV-negative women clearly indicate that sexual intercourse is virtually a necessary step for acquiring HPV. As with any other sexually transmitted disease (STD) men are implicated in the epidemiological chain of the infection. Penile HPVs are predominantly acquired through sexual contacts. Sexual contacts with women who are prostitutes play an important role in HPV transmission and in some populations sex workers may become an important reservoir of high-risk HPVs. Acting both as "carriers" and "vectors" of oncogenic HPVs male partners may markedly contribute to the risk of developing cervical cancer in their female partners. Thus, in the absence of screening programs, a woman's risk of cervical cancer may depend less on her own sexual behavior than on that of her husband or other male partners. Although more rarely than women, men may also become the "victims" of their own HPV infections as a fraction of infected men are at an increased risk of developing penile and anal cancers. Male circumcision status has been shown to reduce the risk not only of acquiring and transmitting genital HPVs but also of cervical cancer in their female partners. More research is needed to better understand the natural history and epidemiology of HPV infections in men. This paper is available too at: http://www.insp.mx/salud/index.html. PMID- 14746028 TI - Cervical carcinogenesis: the role of co-factors and generation of reactive oxygen species. AB - Several HPV co-factors have been proposed, some more or less consistently associated with cervical dysplasia and cancer risk. More research, using prospective cohort designs, is needed to further describe where in carcinogenesis these factors are working and to assess the biological mechanism of these factors. In addition, further research is needed to define the role of various hormonal contraceptive formulations in promoting cervical carcinogenesis. While many interesting scientific questions remain to be answered, results from the numerous epidemiological studies conducted to date indicate that cervical dysplasia and cancer may be reduced if the oxidant antioxidant ratio is shifted to more of and antioxidant profile. In addition to cervical cancer screening, a reduction in cervical cancer incidence may be accomplished by reducing tobacco use, increasing nutritional status, and utilizing barrier contraception to prevent infection with other sexually acquired infections. This paper is available too at: http://www.insp.mx/salud/index.html. PMID- 14746029 TI - Serology for human papillomavirus. AB - Difficulties with serology for papillomavirus are associated with the large number of human papillomavirus, cross-reactions between papillomavirus, and to the diversity of lesions and target sites for infection. In addition, the expression of the papillomavirus in the superficial layers of the epithelium gives rise to the weak presentation to immunocompetent cells of viral antigens, which in turn gives rise to a weak serological response. Distinct efforts have been made in previous decades to develop more specific and sensitive serological assays. These former studies use fusion proteins and synthetic peptides, although they remain on the whole uninteresting, due to their lack of sensitivity and specificity. Only in the last few years, and principally due to the advent of various virus-like particles (VLP), have more sensitive and specific assays become available. This paper is available too at: http://www.insp.mx/salud/index.html. PMID- 14746030 TI - Prospects for controlling cervical cancer at the turn of the century. AB - Cervical cancer morbidity and mortality have decreased substantially during the last 50 years mostly due to successful organized or opportunistic screening with Pap cytology in high and middle income countries. In many low income countries Pap cytology screening is yet to be effectively implemented or has failed to reduce cervical cancer rates to an appreciable extent. The fact that infection with certain human papillomavirus (HPV) types is now recognized as a necessary cause of this disease has led to new research fronts on prevention of cervical cancer. Testing for HPV DNA has shown great promise as a screening tool with better sensitivity but somewhat lower specificity than Pap cytology. In combination with the latter, HPV testing has the potential to improve the negative predictive value of cytology, thus allowing for increased testing intervals, which would lower program costs with acceptable safety. Advances in cytology processing and automation have also led to new screening approaches that are increasingly gaining acceptance in high and middle income countries. For low income countries, visual inspection with acetic acid has proven to be an effective alternative to conventional Pap cytology, especially in settings where no screening programs have been implemented. Concerning primary prevention of cervical cancer, recent research on the safety and efficacy of candidate prophylactic vaccines against HPV have shown very promising results with nearly 100% efficacy in preventing persistent infections and development of cervical cancer precursors. However, policy makers are strongly cautioned to avoid deferring decisions concerning the implementation of cervical cancer screening under the expectation that a successful vaccine could obviate the need for secondary prevention strategies. This paper is available too at: http://www.insp.mx/salud/index.html. PMID- 14746031 TI - Screening for cervical cancer: new alternatives and research. AB - Evidence for the clinical utility of human papillomavirus (HPV) DNA testing has increased over the years and has now become very convincing. Some specific uses of HPV detection are a) triage of women with cytological determinations of atypical squamous cells of undetermined significance (ASC-US) and related management strategies, b) as a marker for test of cure post-treatment, and c) most importantly, as an adjunct to cytology in routine cervical disease screening programs. There are many studies that support each of these applications and include 8 studies on ASC-US triage, 10 on test of cure and 13 on adjunctive or stand-alone HPV screening. The most notable investigation of ASC-US triage was ALTS, a randomized controlled trial of 3,488 women. With respect to routine HPV screening the combined studies included 77,000 women, providing as a histological endpoint more than 1,000 cases of high-grade cervical intraepithelial neoplasia (CIN) or cancer. Testing methods were either the Hybrid Capture 2 (HC2) test or the polymerase chain reaction (PCR) test. HPV testing of women with ASC-US cytology had on average a higher sensitivity (90%) and specificity (70%) than repeating the cytological test (sensitivity 75%, specificity 60%) and was also more sensitive than colposcopy for follow-up. As an adjunct to the Papanicolaou (Pap) cytology test in routine screening, HPV DNA testing was a more sensitive indicator for prevalent high-grade CIN than either conventional or liquid cytology. A combination of HPV DNA and Papanicolaou testing had almost 100% sensitivity and negative predictive value. The specificity of the combined tests was slightly lower than the specificity of the Papanicolaou test. One "double negative" HPV DNA and Papanicolaou test indicated a higher prognostic assurance against risk of future CIN 3 than three subsequent negative conventional Papanicolaou tests and may safely allow three-year or longer screening intervals for such low-risk women. It appears that HPV DNA testing is on the way to becoming a common testing strategy in cervical cancer prevention programs. Research continues into approaches for improving the performance and cost effectiveness of HPV detection methods. Hybrid Capture 3 will offer improved HPV typing capabilities and the Rapid Capture machine allows for robot-assisted HPV DNA testing, permitting greater test throughput. PCR test improvements are expected to contribute to the growth of flexible accurate and cost-effective HPV DNA tests. It is likely that improved diagnostic technology along with HPV genotyping and quantitation may provide more value in future. A particularly promising approach is to combine HPV DNA testing with expression levels of other markers such as proliferative or cell cycle regulatory proteins to subdivide HPV positive women into those who are at greater risk of cancer and those who can be safely followed by screening at longer intervals. This paper is available too at: http://www.insp.mx/salud/index.html. PMID- 14746032 TI - Improving cervical cancer screening in Mexico: results from the Morelos HPV Study. AB - OBJECTIVE: The purpose of this paper is to describe some of the results of the Morelos HPV Study. The main objective of the Morelos HPV Study is to evaluate the use of human papillomavirus (HPV) DNA testing, as compared to the Papanicolaou (Pap) test, for cervical cancer (CC) screening. MATERIAL AND METHODS: The Morelos HPV Study is currently being conducted in Mexico, to examine the possibility of using HPV testing for CC screening. The HPV testing of self-collected vaginal and clinician-collected cervical specimens was evaluated as part of this study. The acceptability of the HPV testing of self-collected specimens was compared to that of the Pap test. A cost-effectiveness analysis (CEA) and cost-benefit analysis (CBA) was also performed. RESULTS: The Morelos HPV Study results indicate that HPV testing has a greater sensitivity to detect cervical intraepithelial neoplasia (CIN) 2/3 and CC than the Pap test. Our results also indicate an over all lower acceptability of the Pap test as compared to the self-collected procedure. The results of the CEA and CBA indicate that screening women between the ages of 20-80 for CC using some type of HPV testing is always more cost effective than screening for CC using the Pap test. CONCLUSIONS: Our results suggest that self- and clinician-collected HPV testing could be used in CC prevention programs, as an effective complement or substitute for the Pap test. This paper is available too at: http://www.insp.mx/salud.index.html. PMID- 14746033 TI - Early detection of cervical cancer with visual inspection methods: a summary of completed and on-going studies in India. AB - India is a high-risk country for cervical cancer which accounts a quarter (126,000 new cases, 71,000 deaths around 2,000) of the world burden. The age standardized incidence rates range from 16-55 per 100,000 women in different regions with particularly high rates in rural areas. Control of cervical cancer by early detection and treatment is a priority of the National Cancer Control Programme of India. There are no organized cytology screening programmes in the country. The technical and financial constraints to organize cytology screening have encouraged the evaluation of visual inspection approaches as potential alternatives to cervical cytology in India. Four types of visual detection approaches for cervical neoplasia are investigated in India: a) naked eye inspection without acetic acid application, widely known as 'downstaging'; b) naked eye inspection after application of 3-5% acetic acid (VIA); c) VIA using magnification devices (VIAM); d) visual inspection after the application of Lugol's iodine (VILI). Downstaging has been shown to be poorly sensitive and specific to detect cervical neoplasia and is no longer considered as a suitable screening test for cervical cancer. VIA, VIAM and VILI are currently being investigated in multicentre cross-sectional studies (without verification bias), in which cytology and HPV testing are also simultaneously evaluated, and the results of these investigations will be available in 2003. These studies will provide valuable information on the average, comparative test performances in detecting high-grade cervical cancer precursors and cancer. Results from pooled analysis of data from two completed studies indicated an approximate sensitivity of 93.4% and specificity of 85.1% for VIA to detect CIN 2 or worse lesions; the corresponding figures for cytology were 72.1% and 91.6%. The efficacy of VIA in reducing incidence of an mortality from cervical cancer and its cost effectiveness is currently being investigated in two cluster randomized controlled intervention trials in India. One of these studies is a 4-arm trial addressing the comparative efficacy of VIA, cytology and primary screening with HPV DNA testing. This trial will provide valuable information on comparative detection rates of CIN 2-3 lesions by the middle of 2003. The expected outcomes from the Indian studies will contribute valuable information for guiding the development of public health policies on cervical cancer prevention in countries with different levels of socio-economic and health services development and open up new avenues of research. This paper is available too at: http//www.insp.mx/salud/index.html. PMID- 14746035 TI - Cervical squamous and glandular intraepithelial neoplasia: identification and current management approaches. AB - Certain types of human papillomaviruses (HPV) are associated with squamous intraepithelial lesions and cancer and these are termed high-risk. HPV type 16 is detected in approximately half of the high-grade squamous intraepithelial lesions and cancer. Because of the high rate of spontaneous regression of low-grade squamous lesions, follow-up by cytology, colposcopy and possible biopsy appears preferable. Due to the higher rate of progression to malignancy of the high-grade lesions conservative treatment is recommended. One of the most common reasons for persistence relates to the human immunodeficiency virus. Adenocarcinoma in situ is an uncommon disorder and not well identified by cytologic sampling or colposcopic inspection. The diagnosis is made by cone biopsy, the specimen having negative margins for disease. Hysterectomy is the treatment procedure of choice unless fertility is an issue. Excisional methods (particularly electrosurgical loop) can interfere with accurate histological interpretation in some cases of both squamous disease and adenocarcinoma in situ. This paper is available too at: http://www.insp.mx/salud/index.html. PMID- 14746034 TI - Factors affecting utilization of cervical cancer prevention services in low resource settings. AB - Strategies for introducing or strengthening cervical cancer prevention programs must focus on ensuring that appropriate, cost-effective services are available and that women who most need the services will, in fact, use them. This article summarizes the experiences of research projects in Bolivia, Peru, Kenya, South Africa, and Mexico. Factors that affect participation rates in cervical cancer prevention programs are categorized in three sections. The first section describes factors that arise from prevailing sociocultural norms that influence women's views on reproductive health, well being, and notions of illness. The second section discusses factors related to the clinical requirements and the type of service delivery system in which a woman is being asked to participate. The third section discusses factors related to quality of care. Examples of strategies that programs are using to encourage women's participation in cervical cancer prevention services are provided. This paper is available too at: http://www.insp.mx/salud/index.html. PMID- 14746036 TI - Prospects for primary prevention of cervical cancer in developing countries. AB - The HPV types that cause cervical cancer are sexually transmitted, but there is little evidence that infection can be avoided by behavioural changes, such as condom use. In contrast, prophylactic vaccines against HPV infection are likely to have high efficacy. In principle, the effectiveness of HPV vaccination as a strategy for cervical cancer control can be measured either by monitoring secular trends in cervical cancer incidence or by conducting randomized trials. The former approach is unlikely to provide convincing evidence of effectiveness, since cervical cancer rates are subject to strong secular trends that are independent of intervention measures. A few phase III trials of HPV prophylactic vaccines are now being started. Such trials are very expensive studies involving frequent and complicated investigations. It is important, however, to start as soon as possible simpler trials designed to demonstrate the effectiveness of HPV vaccine in field conditions, i.e. in developing or intermediate countries which suffer the major burden of mortality from cervical cancer. Such trials may capture a difference in the most severe, and rarest, preinvasive cervical lesions (i.e., the real target of any HPV vaccine) over a prolonged follow-up (20 years at least). The design of such studies is briefly considered for two areas: Southern India and South Korea. This paper is available at: http://www.insp.mx/salud/index.html. PMID- 14746037 TI - Vaccines against human papillomavirus and perspectives for the prevention and control of cervical cancer. AB - Today, "persistent" infections by certain types of human papillomavirus (HPV) are considered necessary for developing cervical cancer. Producing efficient vaccines against these viruses may eventually lead to a great reduction in incidence and mortality rates of this cancer. In the case of HPV, the production of traditional vaccines usually based in dead or attenuated viruses is not possible due in part to the lack of systems where large quantities of viral particles could be obtained. Fortunately, the expression of the late L1 protein alone, or in combination with L2, leads to the generation of structures resembling true virions that have been called virus-like particles (VLPs) and constitute excellent candidates as prophylactic vaccines. VLPs have shown to be very immunogenic, and have prevented development of natural or challenged infections in both animal systems and humans. Recently, HPV16VLPs were shown to be very efficient to prevent the development of "persistent" infections, as determined by PCR assays, in a large group of vaccinated women. Therapeutic vaccines, on the other hand, are expected to have an impact on advanced lesions and residual illness, by taking advantage of the fact that early E6 and E7 genes are thought to be constitutively expressed in cervical tumors and precursor lesions. Finally, DNA-based vaccines could represent a useful alternative for preventing infections by genital HPV. This paper is available too at: http://www.insp.mx/salud/index.html. PMID- 14746038 TI - Vaccines against papillomavirus infections and disease. AB - Squamous cell carcinoma of the uterine cervix is the second cause of cancer related deaths in women, the higher incidence being observed in developing countries. Infection with oncogenic types of human papillomavirus (HPV) is considered the major risk factor for the development of malignancies in the uterine cervix. However, HPV is considered to be a necessary but not sufficient cause for cervical cancer and, therefore, other factors contribute to the carcinogenic process, both present in the environment and from the host. Studies performed in animals, and more recently in humans, indicate that vaccination against the capsid proteins of the virus can prevent efficiently from infection. Furthermore, therapeutic vaccines are under investigation aiming the regression of papillomavirus induced tumors. The scientific basis for the development of papillomavirus vaccines and present status of clinical trials will be addressed in this chapter. This paper is available too at: http://www.insp.mx/salud/index.html. PMID- 14746039 TI - Recommendations for cervical cancer screening programs in developing countries. The need for equity and technological development. AB - The cervical cancer screening programs (CCSP) have not been very efficient in the developing countries. This explains the need to foster changes on policies, standards, quality control mechanisms, evaluation and integration of new screening alternatives considered as low and high cost, as well as to regulate colposcopy practices and the foundation of HPV laboratories. Cervical cancer (CC) is a disease most frequently found in poverty-stricken communities and reflecting a problem of equity at both levels gender and regional, and this, is not only due to social and economic development inequalities, but to the infrastructure and human resources necessary for primary care. For this reason, the CCSP program must be restructured, a) to primarily address unprivileged rural and urban areas; b) to foster actions aimed at ensuring extensive coverage as well as a similar quality of that coverage in every region; c) to use screening strategies in keeping with the availability of health care services. In countries with a great regional heterogeneity, a variety of screening procedures must be regulated and standardized, including a combination of assisted visual inspection, cervical cytology and HPV detection; d) regional community intervention must be set up to assess the effectiveness of using HPV detection as an strategy in addition to cervical cytology (pap smear); e) the practice of colposcopy must be regulated to prevent the use of it in healthy women at a population level, thus preventing unnecessary diagnosis and treatment which not only are expensive but also causes unnecessary anxiety to women at risk; f) the operation of those clinical laboratories using HPV as a detection strategy must likewise be accredited and regulated and g) the CCSP program for assuring health care quality should meet the expectations of its beneficiaries, and increase the knowledge in cervical cancer related matters. Finally, though a variety of clinical tests on prophylactic and therapeutic vaccines against HPV are recently being developed worldwide; it will take at least from 5 to 10-years time to have them available in the market. For this reason, it will be necessary to intensify the CCSP programs. All these reasons lay emphasis on the need to reinforce actions for CCSP programs. This paper is available too at: http://www.insp.mx/salud/index.html. PMID- 14746040 TI - [Results of the Second Nutrition National Survey]. PMID- 14746041 TI - Nutritional status of indigenous children younger than five years of age in Mexico: results of a national probabilistic survey. AB - OBJECTIVE: To compare the prevalence of undernutrition and anemia in indigenous and non-indigenous children < 5 years of age at the national level, by region and by urban and rural areas, and to evaluate the degree to which the socioeconomic condition of the family predicts the differences. MATERIAL AND METHODS: A national probabilistic survey was conducted in Mexico in 1999. Indigenous families were identified as those in which at least one woman 12-49 years of age in the household spoke a native language. The prevalence of undernutrition (stunting, wasting and underweight) and anemia was compared between indigenous and non-indigenous children. Probability ratios (PR) were used to compare prevalences in indigenous and non-indigenous children adjusting for socioeconomic status (SES) of the family and for other covariates. RESULTS: The prevalences of stunting and underweight were greater in indigenous than in non-indigenous children. At the national level and in urban areas the prevalences were three times greater and in rural areas approximately 2 times greater (p < 0.05). No differences were found in the prevalence of wasting (p > 0.05). The prevalence of anemia in indigenous children was one third greater than in non-indigenous children at the national level (p < 0.05) and was between 30 and 60% greater in urban areas and in the regions studied (p < 0.05) but was not statistically significant (p > 0.05) in rural areas. These differences were reduced to about half when adjusting for SES but remained significantly higher in indigenous children (p < 0.05). CONCLUSIONS: Indigenous children have higher probabilities of stunting and underweight than non-indigenous children. The differences are larger in urban areas and in higher socioeconomic geographic regions and are explained mostly by socioeconomic factors. The overall difference in the probability of anemia is small, is higher only in urban relative to rural areas, and is explained to a lesser degree by socioeconomic factors. Policy and programs should be designed and implemented to reduce the dramatic differences in nutritional status between indigenous and non-indigenous children in Mexico. The English version of this paper is available too at: http://www.insp.mx/salud/index.html. PMID- 14746042 TI - Breast-feeding practices in Mexico: results from the Second National Nutrition Survey 1999. AB - OBJECTIVE: To assess breast-feeding (BF) practices and determinants of exclusive BF (EBF) < 4 and < 6 months (mo) among women and infants < 23 mo in the NNS-1999. MATERIAL AND METHODS: BF practices from the day and night before the interview were ascertained, and median duration estimated. Determinants of EBF < 4 and < 6 mo were analysed by logistic regression models for complex surveys. RESULTS: Prevalence of EBF < 4 mo was 25.7%, and of < 6 mo 20.3%. The overall rate of continued BF (second year) was 30.9%, median duration of BF 9 mo, and the national proportion of children ever breast-fed 92.3%. The probability (p) of EBF < 4 mo was determined by infant age and sex, by maternal socio-economic level (SEL) and ethnicity, and by the interaction between infant sex and SEL. The pEBF < 6 mo was determined by infant age and length, by maternal ethnicity, and employment. CONCLUSIONS: EBF rates and duration are low in Mexico and have improved only slightly in the last 20 y. Infant and maternal characteristics determine the pEBF. If improvements in infant health are a national priority, aggressive interventions to promote and protect BF are urgently needed in Mexico, as well as formal evaluation of current initiatives. The English version of this paper is available too at: http://www.insp.mx/salud/index.html. PMID- 14746043 TI - Prevalence of anemia in children 1 to 12 years of age. Results from a nationwide probabilistic survey in Mexico. AB - OBJECTIVE: To describe the epidemiology and analyze factors associated with iron deficiency anemia in a probabilistic sample of the Encuesta Nacional de Nutricion 1999 (ENN-99) [National Nutritional Survey 1999 (NNS-99)]. MATERIAL AND METHODS: The sample included 8,111 children aged 1 to 12 years, and was nationally representative by rural and urban strata and by four geographical regions. Capillary hemoglobin was measured using a portable photometer (HemoCue). The analysis of the determining factors of anemia was performed by odds ratios derived from a logistic regression model and multiple regression models. RESULTS: The prevalence of anemia was 50% in infants < 2 years of age, with no significant differences between urban and rural strata or among regions. It varied between 14 and 22% in 6-11 year-old children and was higher in the South region and among the indigenous children. Dietary intake of iron was 50% of the recommended daily allowance in children < 2 years of age, but not in older children. Phytate ( approximately 500-800 mg/d) and tannin (approximately 19 mg/d) intakes were very high in children over 7 years of age. Hemoglobin was positively associated with nutritional status of children (p = 0.01), socioeconomic status (p range 0.05 0.001), duration of lactation in children under 2 years of age (p = 0.1), and iron and calcium intake (p = 0.02), but not with folic acid or vitamin B12 intake. Hemoglobin was negatively associated with maternal education (p = 0.01) in older children, but not in those under 2 years of age. CONCLUSIONS: We present evidence of an alarming national epidemic of anemia, particularly marked in children 12 to 24 months of age. The control of anemia should be considered as an urgent national concern given its grave consequences on the physical and mental development of these children and on their long-term health. The English version of this paper is available too at: http://www.insp.mx/salud/index.html. PMID- 14746044 TI - Anemia in Mexican women: a public health problem. AB - OBJECTIVE: The purpose of this study is to quantify the prevalence and distribution of anemia among women of childbearing age (12 to 49 years) participating in the 1999 National Nutrition Survey (NNS-1999). MATERIAL AND METHODS: The survey had a probabilistic design and was representative at the national level, of urban and rural areas and four regions: North, South, Center, and Mexico City. Hemoglobin concentration was determined in capillary blood samples using a portable photometer (HemoCue), in 17,194 women, 697 of whom were pregnant. RESULTS: The overall prevalence of anemia was 27.8% in pregnant women and 20.8% in non-pregnant women. Higher prevalences were observed in rural as compared to urban areas, both in pregnant (28.0% vs 27.7%) and non-pregnant (22.6% vs 20.0%) women, but the differences were not statistically significant (p > 0.05). Women in the South had the greatest prevalence (23.2%), followed by those in the North (20.9%), Center (20.6%), and Mexico City (16.4%). Non-pregnant indigenous women had a prevalence of 24.8%, while in-non-indigenous women the prevalence was 20.4%. CONCLUSIONS: Anemia in women of childbearing age is a growing public health problem that justifies the implementation of interventions for its prevention and control. The English version of this paper is available too at: http://www.insp.mx/salud/index.html. PMID- 14746045 TI - Vitamins A, and C and folate status in Mexican children under 12 years and women 12-49 years: a probabilistic national survey. AB - OBJECTIVE: To describe the epidemiology of Vitamin A and C and folic acid deficiencies and their association with sociodemographic and dietary factors in a national probabilistic sample of Mexican women and children. MATERIAL AND METHODS: This is a probabilistic sample from the National Nutrition Survey 1999 (ENN-99) including 1,966 children and 920 women. Vitamins A and C were measured in serum by high-performance liquid chromatography, and folic acid in total blood by a microbiological method. Determinants for such deficiencies were explored by multiple regression models. RESULTS: Vitamin A deficiency (retinol < 10 micrograms/dl) was rare in both children and women. But subclinical deficiency (retinol > 10 and < 20 micrograms/dl) was present in 25% of children. The likelihood of subclinical deficiency of vitamin A was less in older children (OR = 0.98, p = 0.01) and in women with higher body mass index (OR = 0.93, p = 0.01). About 30% of children < 2 years of age and 40% of women were vitamin C deficient. The likelihood of vitamin C deficiency was less in children and women as socioeconomic level increased (OR = 0.69, p = 0.03, and OR = 0.80, p = 0.04), and higher in older women (OR = 1.02, p = 0.05). The prevalence of folate deficiency varied in children (2.3 to 11.2), in women it was 5%. Folate deficiency was less in children of higher socioeconomic level (OR = 0.62, p = 0.01), and in those eating more vegetables (OR = 0.22, p = 0.01). CONCLUSIONS: The high prevalence of subclinical deficiency of vitamin A in children is indicative of risk of further deterioration under adverse circumstances. Vitamin C deficiency in both children and women implies in addition diminished ability for iron absorption. The English version of this paper is available too at: http://www.insp.mx/salud/index.html. PMID- 14746046 TI - Iron, zinc and iodide status in Mexican children under 12 years and women 12-49 years of age. A probabilistic national survey. AB - OBJECTIVE: To describe the epidemiology of iron, zinc and iodide deficiencies in a probabilistic sample of Mexican women and children and explore its association with some dietary and socio-demographic variables. MATERIAL AND METHODS: We carried out in 1999 an epidemiological description of iron (percent transferrin saturation, PTS, < 16%), serum zinc (< 65 ug/dl) and iodide (< 50 ug/l urine) deficiencies in a probabilistic sample of 1,363 Mexican children under 12 years and of 731 women of child-bearing age. Serum iron, Total Iron Binding Capacity (TIBC) and zinc were measured by atomic absorption spectrometry, and urinary iodide by a colorimetric method. Logistic regression models explored determinants for such micromineral deficiencies. RESULTS: Iron deficiency was higher (67%) in infants < 2 years of age. Prevalence declined (34-39%) at school age. The prevalence for iron deficiency in women was 40%. Zinc deficiency was higher in infants < 2 years of age (34%) than in school-age children (19-24%). Prevalence in women was 30%, with no rural/urban difference. In women the likelihood of iron deficiency decreased as SEL improved (p = 0.04) and increased with the intake of cereals (p = 0.01). The likelihood of low serum zinc levels was greater in women and children of low socioeconomic level (SEL) (p < 0.02 and p = 0.001) iodide deficiency was negligible in both children and women. CONCLUSIONS: The data shows high prevalence of iron deficiency-specially in infants 12 to 24 months of age. It is suggested that in older children and women 12 to 49 years of age that iron bioavailability is low. The prevalence of zinc deficiency was also very high. The English version of this paper is available too at: http://www.insp.mx/salud/index.html. PMID- 14746047 TI - Energy and nutrient consumption in Mexican women 12-49 years of age: analysis of the National Nutrition Survey 1999. AB - OBJECTIVE: To describe the reported energy and nutrient intake and adequacies in Mexican women. MATERIAL AND METHODS: A 24-hour dietary recall was used to obtain nutrient intake in a representative sub-sample of 2,630 women from 12 to 49 years of age from the National Nutrition Survey 1999. Nutrient adequacies were estimated using the Dietary Reference Intakes and stratified according to region, area (urban or rural), socioeconomic status and obesity status (non-obese: BMI < 30 kg/m2, obese: > or = 30 kg/m2). Differences were analyzed using linear regression for complex surveys of log-transformed intake and adequacy, adjusting for multiple comparisons with the Bonferroni test. RESULTS: The median national energy intake was 1,471 kcal. The Risk of Inadequacy (RI) (prevalence of adequacy < 50%) was: vitamin A: 38.3%, vitamin C: 45.5%, and folate: 34.3%. Carbohydrates, folate, iron and calcium intake was significantly higher in rural than in urban areas. The RI was higher in women of the lowest socioeconomic status tertile for all nutrients with the exception of carbohydrates and calcium. Macro-nutrient adequacies were significantly higher in non-obese women. CONCLUSIONS: Differences within the country among regions, rural and urban areas, and socioeconomic status tertile reflect an increasing availability of inexpensive calorie-dense foods in marginal groups. However, total energy, cholesterol, saturated and total fat were consumed in greater quantities by women from the higher socioeconomic status tertile and from urban areas. These patterns could be a contributing factor to the rise of obesity and other non-communicable nutrition-related chronic diseases in Mexico. The English version of this paper is available at: http://www.insp.mx/salud/index.html. PMID- 14746048 TI - Energy and nutrient intake in preschool and school age Mexican children: National Nutrition Survey 1999. AB - OBJECTIVE: To estimate energy and nutrient intake and adequacy in preschool and school age Mexican children, using the National Nutrition Survey 1999 (NNS-1999). MATERIAL AND METHODS: Twenty four-h dietary recalls from pre-school (n = 1,309) and school (n = 2,611) children obtained from a representative sub-sample of the NNS-1999 were analyzed. Intakes and adequacies were estimated and compared across four regions, socio-economic strata, and between urban and rural areas, and indigenous vs. non-indigenous children. RESULTS: Median energy intake in pre school children was 949 kcal and in school children 1,377 kcal, with adequacies < 70% for both groups. Protein adequacy was > 150% in both age groups. The North and Mexico City regions had the highest fat intake and the lowest fiber intake. Children in the South region, indigenous children, and those in the lowest socio economic stratum had higher fiber and carbohydrate intakes and the lowest fat intake. These children also showed the highest risks of inadequacies for vitamin A, vitamin C, folate, iron, zinc and calcium. CONCLUSIONS: Mexico is experiencing a nutrition transition with internal inequalities across regions and socio economic strata. Food policy must account for these differences in order to optimize resources directed at social programs. The English version of this paper is available too at: http://www.insp.mx/salud/index.html. PMID- 14746049 TI - Factors associated with overweight and obesity in Mexican school-age children: results from the National Nutrition Survey 1999. AB - OBJECTIVE: The objective of the study was to measure the prevalence of overweight and obesity in Mexican school-age children (5-11 years) in the National Nutrition Survey 1999 (NNS-1999). MATERIAL AND METHODS: Overweight and obesity (defined as an excess of adipose tissue in the body) were evaluated through the Body Mass Index (BMI) in 10,901 children, using the standard proposed by the International Obesity Task Force. Sociodemographic variables were obtained using a questionnaire administered to the children's mothers. RESULTS: The national prevalence of overweight and obesity was reported to be 19.5%. The highest prevalence figures were found in Mexico City (26.6%) and the North region (25.6%). When adjusting by region, rural or urban area, sex, maternal schooling, socioeconomic status, indigenous ethnicity and age, the highest prevalences of overweight and obesity were found among girls. The risks of overweight and obesity were positively associated with maternal schooling, children's age and socioeconomic status. CONCLUSIONS: Overweight and obesity are prevalent health problems in Mexican school-age children, particularly among girls, and positively associated with socioeconomic status, age, and maternal schooling. This is a major public health problem requiring preventive interventions to avoid future health consequences. The English version of this paper is available too at: http://www.insp.mx/salud/index.html. PMID- 14746050 TI - Methods of the National Nutrition Survey 1999. AB - OBJECTIVE: To describe the methods and analyses of the 1999 National Nutrition Survey (NNS-99). MATERIAL AND METHODS: The 1999 National Nutrition Survey (NNS 99) is a probabilistic survey with nationwide representativity. The NNS-99 included four regions and urban and rural areas of Mexico. The last sampling units were households, selected through stratified cluster sampling. The study population consisted of children under five years of age, school-age children (6 11 years), and women of childbearing age (12-49 years). Data were collected on anthropometric measurements, hemoglobin levels, morbidity and its determinants, and socioeconomic and demographic characteristics. In addition, data on diet and micronutrients intakes (iron, zinc, vitamin A, folic acid, vitamin C, and iodine) were obtained in a sub-sample of subjects. RESULTS: The response rate for the NNS 99 was 82.3%; the non-response rate was 5.9% and the remaining did not participate due to uninhabited houses. CONCLUSIONS: This survey updates the information on nutritional status in Mexico and should serve as the basis for food and nutrition policy-making and priority program design. The English version of this paper is available too at: http://www.insp.mx/salud/index.html. PMID- 14746051 TI - Conclusions from the Mexican National Nutrition Survey 1999: translating results into nutrition policy. AB - OBJECTIVE: This article presents and overview of the main results and conclusions from the Mexican National Nutrition Survey 1999 (NNS-1999) and the principal nutrition policy implications of the findings. MATERIAL AND METHODS: The NNS-1999 was conducted on a national probabilistic sample of almost 18,000 households, representative of the national, regional, as well as urban and rural levels in Mexico. Subjects included were children < 12 years and women 12-49 years. Anthropometry, blood specimens, diet and socioeconomic information of the family were collected. RESULTS: The principal public nutrition problems are stunting in children < 5 years of age; anemia, iron and zinc deficiency, and low serum vitamin C concentrations at all ages; and vitamin A deficiency in children. Undernutrition (stunting and micronutrient deficiencies) was generally more prevalent in the lower socioeconomic groups, in rural areas, in the south and in Indigenous population. Overweight and obesity are serious public health problems in women and are already a concern in school-age children. CONCLUSIONS: A number of programs aimed at preventing undernutrition are currently in progress; several of them were designed or modified as a result of the NNS-1999 findings. Most of them have an evaluation component that will inform adjustments or modifications of their design and implementation. However, little is being done for the prevention and control of overweight and obesity and there is limited experience on effective interventions. The design and evaluation of prevention strategies for controlling obesity in the population, based on existing evidence, is urgently needed and success stories should be brought to scale quickly to maximize impact. The English version of this paper is available too at: http://www.insp.mx/salud/index.html. PMID- 14746052 TI - [Malnutrition]. PMID- 14746053 TI - [Enzootic calcinosis in sheep after consumption of golden oat grass (Trisetum flavescens L., P. B.)]. AB - In 12 sheep (Coburger Fuchsschaf; 10 female, 1 male, 1 wether), grazing on a pasture with approximately 20 (to 40)% golden oat grass and fed the respective hay, clinical findings and outcome of Trisetum flavescens induced calcinosis were controlled for 2 1/2 years. Besides lameness typical for calcinosis (slightly bent carpus and relatively stretched position of tarsal and fetlock joints when standing, 'kneeling' on the carpi) the patients showed increasing impairment of the circulatory and respiratory systems (holosystolic endocardial murmur, congestion, exspiratory dyspnoea etc.). 4 of the 12 sheep died (2) or had to be euthanatized (2) because of peracute heart failure; 5 patients showing chronic circulatory insufficiency as well as 1 animal suffering from severe lameness had likewise to be euthanatized (Ubersicht 1-3). One calcinotic ewe (No. 12), still alive at the time of this evaluation (2003), gave birth to 2 healthy lambs and nursed them. The ram (No. 5) had to be eliminated for another disease. The mostly severe calcifications of the cardiac valves, the endocardium and the arterial vessels as observed during the pathomorphological examination are consistent with the clinically diagnosed cardiovascular insufficiency. Furthermore, calcification of several tendons and ligaments, the kidneys and in 3 cases of the pulmonary parenchyma could be found. Clinical observations and post mortem findings showed a remarkable individual variation. Compared to calcinosis in cattle, in sheep the functional disturbance of the circulatory system was striking. PMID- 14746054 TI - [The incidence of cervical dystocia and disorders of cervical involution in the post partum cow]. AB - The aim of this study was to obtain information on the frequency of cervical dystocia and involution disorders of the canalis cervicalis after difficult dystocia and to characterise factors possibly influencing these pathological conditions. Therefore 317 difficult births in the cow (extraction: 123; foetotomy: 82; caesarean section: 112) and the involution of the cervical canal were documented during the first ten days post partum. In total a cervical dystocia could be diagnosed 53 times (16.7% related to the total number of births). This kind of birth disorder is regularly followed by a caesarean section (p < 0.001). Animals who show a narrowness in the cervix were older than the cows without cervical dystocia (p < 0.05). The same correlation could be detected for the number of births. Animals with delivery problems associated with the cervix had already given birth to more calves than cows who were not affected by cervical disorders (p < 0.01). 58 cows developed a disorder of the cervical involution (18.3% related to the total number of deliveries). This puerperal disorder can be frequently observed after foetotomy (p < 0.001). A correlation between the incidence of disorders of cervical involution and the age or the parity of the animals could not be detected. In the same way, cows with a diagnosis of a cervical dystocia did not develop more frequently disorders of cervical involution than animals who showed a physiological dilation of the cervical canal intra partum. PMID- 14746055 TI - [Airborne microorganisms in a rearing henhouse for layers during vaccination]. AB - Airborne microorganisms are proved regularly in livestock houses as a part of stable dust and its amount depends on housing conditions, the flow of air and the movement of material. Health of animals and farmers can be influenced in a negative way by these bioaerosols. In a rearing house for layers concentrations of various groups of airborne microorganisms were measured during vaccination by a veterinary and his three assistants. During the vaccination activities the concentrations of some airborne microorganisms increased by a factor of ten to the following medians of colony forming units (cfu) on used selective agars (cfu/m3): 10(3) on MacConkey (36 degrees C), 10(3) on Dichloran-Glycerol (25 degrees C), 10(7) on Tryptone Soy (CaSo, 36 degrees C), 10(3) on Salmonella Shigella (36 degrees C), 10(2) yeasts on Sabouraud (36 degrees C), and 10(2) on Campylobacter (36 degrees C). Thermophilic fungi were only grown on some of the used Maltextract agar dishes (45 degrees C) in concentrations near of the limit of detection. Some aerial samples were analysed for Chlamydia. Chlamydophila psittaci was not detected. Concentrations of airborne microorganisms in livestock houses depends not only on housing conditions but also on specific work procedures of farmers or on the activity of the animals. PMID- 14746056 TI - A smart system for surveillance of animal welfare during transport. AB - New welfare regulations will impose surveillance systems so that information on the quality of transport conditions is available. Moreover a route description is useful for optimisation of transport logistics, but also in relation to estimating of sanitary risk and food safety, including traceability of individual animals. Therefore a transport surveillance system has been developed which is integrating the following information: individual identification of animals, (un)loading place and time, air quality (temperature, relative humidity, emissions), vibration and behaviour of the animals. These data are collected by telemetry and GPS, and are transmitted to a dispatch centre by GSM. Hence, information is available on-line and on disk, so that the driver can be informed and corrected at the spot. Dynamic route optimization of cattle collection from farms and logistical activities of abattoirs are considered in relation to animal welfare. Another instrumentation package that comprises sensors of heart rate and vibration on the animal has been integrated. These sensors can be mounted on animals and the data is transferred to a database through a wireless network. Comprehensive field measurement has been made to evaluate the system and found that the package performs well. Hence, advice will be generated for vehicle manufacturers, hauliers, farmers, slaughterhouses and retailers. PMID- 14746057 TI - [Differences in the seroprevalence of Salmonella spp. in free-ranging and corralled semi-domesticated reindeer (Rangifer tarandus tarandus) in Finland]. AB - Serum samples from 1032 semi-domesticated reindeer (Rangifer tarandus tarandus) from Finland were examined for the occurrence of Salmonella-antibodies by use of an indirect ELISA. The majority of samples originated from clinically healthy slaughter reindeer, kept extensively (n = 802; year of sampling: 1996). The remaining samples (n = 230) came from a research herd, permanently kept intensively, with repeated outbreaks of diarrhoea. In this study, 29 of the examined serum samples showed an OD above the determined cut-off. The prevalence in the clinically healthy slaughter reindeer was 0.9%, in the research herd 4.2% in 1996, 10.5% in 1997 and 12.9% in 1998. It must be assumed that the intensive husbandry in the corralled research herd may favour the spreading of infectious agents and eventually outbreaks of crowding diseases in the herd. This investigation is complemented by a review on the occurrence of Salmonella in wild and semi-domesticated cervids. PMID- 14746058 TI - [B-mode and color Doppler sonography of kidneys in healthy lambs and lambs with urolithiasis]. AB - The objectives of this study were to determine some morphological characteristics of kidneys and resistive index (RI) changes in renal interlobar artery using B Mode and color coded doppler sonography in lambs with urolithiasis. The study was performed in 14 male lambs (8 patients, 6 healthy) aged between 2-2.5 months in the same herd. The kidneys had a mean length of 55.6 +/- 1.6 mm, a width of 31.8 +/- 1.2 mm and a parenchymal width of 10.6 +/- 0.6 mm in healthy lambs. These values were 95.4 +/- 3.6., 52.4 +/- 1.3 and 8.75 +/- 0.5 mm, respectively in lambs with urolithiasis. These differences between both groups were significant. Furthermore, the mean resistive index values in lambs with urolithiasis (0.81 +/- 0.017 m/sn) were significantly higher than controls (0.62 +/- 0.013 m/sn). It was concluded that B-Mode and color coded doppler sonography might provide useful information for detection of changes in kidneys of lambs with urolithiasis like hydronephrosis and renal swelling as well as elevated resistance in the renal interlobar artery. PMID- 14746059 TI - [Cross-contamination of ruminant feed stuffs in Bavaria after the comprehensive feed ban]. AB - The results of the official feed control program in Bavaria between 1997 and 2002 document that the proportion of ruminant feedstuffs that was cross-contaminated with animal material was distinctively reduced after the comprehensive feed ban since 2001, and decreased close to zero in 2002. The results underline the high efficiency of the comprehensive feed ban. They also indicate risks of a potential re-admittance of animal protein for nonruminant domestics under the momentary production procedures. PMID- 14746060 TI - Ethical issues in pediatric genetics: views of geneticists, parents and primary care physicians. PMID- 14746061 TI - Children and genetics: comparative study of international policy positions. PMID- 14746062 TI - Selected legal and ethical issues relevant to pediatric genetics. PMID- 14746063 TI - Genetic testing, legal capacity and adolescents. PMID- 14746064 TI - The legal challenge of health research involving children. PMID- 14746065 TI - The mentally incompetent and medical/drug research experimentation: New York saves the day for the underdog. PMID- 14746066 TI - Freebies for subject 641: a discussion of the ethical prospect of providing drug trial subjects with post-trial access to the drug tested--a Canadian perspective. AB - "Pharmaceutical Company Incorporated" is developing a new drugs to treat AIDS. The Company is hoping to begin clinical trials soon and has submitted its tiral proposal for consideration by the Research Ethics Board (REB). The proposal includes provision to supply the drug free of charge to trial subjects if the drug is shown to be effective and safe. The members of the REB are split as to whether such an offer is acceptable. Some members of the REB are concerned that the offer is an undue inducement for prospective subjects to participate in the clinical trial. PMID- 14746067 TI - [The right to health care services under Quebec law]. AB - The main goal of the Canada Health Act is to guarantee that Canadian residents have reasonable access to a comprehensive and universal health care plan. However, reduced federal funding for health care and increases in health care costs due to technical and scientific developments have created unprecedented financial pressures on provincial health care systems. The right to health care, once perceived as one of the pillars of Canadian society, may be imperiled. This article will provide a detailed analysis of the nature and scope of the right to health care from mainly a legal, but also from a political, perspective. Based on the premises that the Canada Health Act is basically a financial agreement between the Federal and provincial governments and that it does not enshrine a substantive right on which individuals may claim services, the author explores the nature and scope of this right under Quebec legislation. Indeed, the Quebec Health and Social Services Act has, since the 1960s, included various provisions that establish a right of access to health care services. This right, however, is fraught with regulatory, organizational and financial limits. The first part of this paper examines relevant regulation from an historical perspective, highlighting the relationships between federal and Quebec provincial legislation. In the second part, the author explores exhaustively the principal provisions relevant to the right to health care. This entails the analysis of administrative regulations as well as of the responsibilities of the various provincial, regional, institutional and professional authorities involved. Ultimately, as this study will demonstrate, the availability of health care services depends more on a vague process than on a legal right to health care. This conclusion is further confirmed by the analysis of the adjudication process of patient complaints provided under the Quebec Health and Social Services Act and by the limited case law on the right of access to services. Even though judges are generally favorable to patients claims, court interventions remain ad hoc and a posteriori. But if the right of access to health care seems limited, its scope is nonetheless important. The right to health care acts as an obstacle to the current dismantling of health care services and to government withdrawal from this field. It also places limits on abusive, arbitrary and discriminatory decision-making and obliges public authorities to take into closer consideration patients' rights in formulating its health care policy and budgets. This protective role of the right to health care is currently illustrated by the politicians' insistence that the ongoing health care reform is not affecting the quality or quantity of available services. PMID- 14746068 TI - Wrestling with the limits of law: regulating new reproductive technologies. PMID- 14746069 TI - Expert psychological opinion evidence in the courts. PMID- 14746070 TI - Benefits of atypical antipsychotic medications in the treatment of psychosis. PMID- 14746071 TI - Kansas illegal tobacco sales to minors: excerpts from the Kansas Annual Synar Report. PMID- 14746072 TI - Future of State Mental Health Hospitals Project Steering Committee report. November, 2003 draft II. PMID- 14746073 TI - Precepts of palliative care. "Managing nutrition and hydration while providing palliative care". PMID- 14746074 TI - Likorall. The lift system of the future. PMID- 14746075 TI - I hate you just the way you are: exploring the formation, maintenance, and need for enemies. PMID- 14746076 TI - Parent-child relationships and enmity with peers: the role of avoidant and preoccupied attachment. PMID- 14746077 TI - Mutual antipathies in the peer group as a moderating factor in the association between community violence exposure and psychosocial maladjustment. PMID- 14746078 TI - Describing the dark side of preadolescents' peer experiences: four questions (and data) on preadolescents' enemies. PMID- 14746079 TI - Enemies in the gendered societies of middle childhood: prevalence, stability, associations with social status, and aggression. PMID- 14746081 TI - Toward understanding mutual antipathies in childhood and adolescence. PMID- 14746080 TI - Developmental risk associated with mutual dislike in elementary school children. PMID- 14746082 TI - [Assessment as a new aspect of nursing insufficiency]. PMID- 14746083 TI - [Scales for evaluation of decubitus ulcer risk]. PMID- 14746084 TI - [The care that nurses give back in return]. PMID- 14746085 TI - [Career profile of nurses]. PMID- 14746086 TI - [Needlestick injuries must not happen]. PMID- 14746087 TI - [It is not right to burden nurses]. PMID- 14746088 TI - [Public health--abundant in Europe]. PMID- 14746089 TI - [Theory and practice in continuing education]. PMID- 14746090 TI - The launch of Shared Visions-New Pathways. PMID- 14746091 TI - Standards review project hits its mark. PMID- 14746092 TI - PFP Clinical/Service Groups and priority focus areas focus survey process. PMID- 14746093 TI - Periodic performance review key to continuous provision of high-quality care. PMID- 14746094 TI - On-site survey brings Shared Vision-New Pathways to the heart of health care organizations. PMID- 14746095 TI - Pilot surveys put new process to the test. PMID- 14746096 TI - Team identifies ways to mitigate risks in new accreditation process. PMID- 14746098 TI - The new JCAHO accreditation report. PMID- 14746097 TI - Accreditation decision process supports continuous operational improvement. PMID- 14746099 TI - Quality report receives additional improvements prior to launch. PMID- 14746100 TI - Security measures protect integrity of Shared Visions-New Pathways system. PMID- 14746101 TI - JCAHO evaluation plan expanded for Shared Visions-New Pathways. PMID- 14746102 TI - The power of one nurse. PMID- 14746103 TI - Attitudes and factors affecting research utilization. AB - PROBLEM: The gap between nursing research findings and their application in practice, particularly in rural areas. METHODS: A descriptive-correlational study design focused on the attitude of nurses (N = 106) in rural practice setting towards nursing research and the relationship between their attitudes and other factors. FINDINGS: Fewer than a quarter of the nurses in this study had favorable attitudes toward research. Attitudes and interest varied with levels of education and position. Also, the isolation of rural nurses from nurse researchers creates a peculiar barrier to research utilization. CONCLUSION: The influence of nurse educators/researchers and administrators cannot be overemphasized. Nurse leaders need to perceive nursing research as the base for evidence-based practice, and enhance the utilization of scientific evidence in practice settings. PMID- 14746104 TI - The suffering of the healer. AB - TOPIC: The severe distress sometimes experienced by nurses in their role as healer. PURPOSE: To identify the sources that give rise to the suffering of the healer, describe the responses of healers to their sufferings, and make recommendations on how to prepare nurses to cope with suffering. SOURCES: The concept of the suffering of the healer is derived from the work of Erik Cassell; the framework for understanding responses to suffering from the work of Dorothy Solle. CONCLUSION: Nurses need to be aware that practice in health care can give rise to the suffering of the healer. Nursing education and administration need to help nurses learn to cope. PMID- 14746105 TI - Clinical practice: an emphasis strategy for promotion and tenure. AB - TOPIC: Emphasizing clinical practice in promotion and tenure materials. PURPOSE: To share a successful strategy centered on clinical practice with members of the tenure track, advanced practice nurse faculty. SOURCE: Published literature, author's experience. CONCLUSION: Creativity and planning can assist tenure track faculty members in emphasizing their clinical practice for promotion and tenure. Recognition and rewards for clinical practice beyond the school of nursing continue to be challenges in the larger univerisity. PMID- 14746106 TI - Concept analysis of adolescent decision making and contraception. AB - TOPIC: Adolescent decision making. PURPOSE: To clarify the concept of adolescent decision making and the use of contraceptives. SOURCE: Published literature. CONCLUSION: The process of adolescent decision making and contraception is influenced by the integration of the intimacy and closeness of the relationship; family, partner and peer influences; locus of control; and self-image. PMID- 14746107 TI - Moving forward way: Medicare prescription drug coverage. AB - Nursing is multidimensional, interactive, interdisciplinary, and complex. Almost anything that can be said about nursing can be said another way. Some things worth being said and heard will not follow the norms of journal presentation. A forum accommodates the emerging voice, the new format, the innovative approach. Nursing Forum, in an effort to honor the independent voice in nursing, presents here the voice who elects to enter the dialogue, but who does so "in another way." PMID- 14746108 TI - Discrimination against the elderly. PMID- 14746109 TI - Laboratory confirmation of a SARS case, southern China. PMID- 14746110 TI - Assessment of neonatal tetanus elimination in Malawi. PMID- 14746111 TI - Recommendations of the Interim Policy on Collaborative TB/HIV activities. PMID- 14746112 TI - Supreme Court rejects attack on medical marijuana. PMID- 14746115 TI - The importance of magnet recognition in recruiting and retaining nurses. PMID- 14746116 TI - Education outreach: taking the "fire" beyond the lecture hall and lab. 2003 Arthur C. Guyton Teacher of the Year Award. PMID- 14746118 TI - Making moments matter. PMID- 14746119 TI - Individual determinants of home-care nursing and housework assistance. AB - The purpose of this study was to examine individual determinants of use of publicly funded home-care nursing and housework assistance by Canadians 18 years and older from 1994 to 1999. Andersen and Newman's Behavioural Model of Health Services Use guided the selection of variables, analyses, and interpretation of the findings. Descriptive, correlation, and multiple logistic regression analyses were completed in each of the first 3 cross-sectional cycles of Statistics Canada's National Population Health Surveys. The determinants of use of housework assistance were older age, female, living alone, lower income, activity restriction, needing help with housework, not hospitalized in the previous year, and having at least 1 chronic condition. The determinants for home nursing tended to be the opposite of those for housework assistance. Between 1994 and 1999, use of housework assistance appeared to decrease and use of nursing services appeared to remain relatively stable. The findings underscore the need to target these 2 discrete subgroups of home-care users and ensure that funding is directed at support services as well as nursing services. PMID- 14746120 TI - Seeking support: caregiver strategies for interacting with health personnel. AB - Support from health professionals can assist family caregivers and have a positive impact on their health. The purpose of this study was to explore women's perceptions of support from community resources while caring for a family member with dementia. The research questions were: What factors influence female caregivers' interactions with health personnel when seeking support? What strategies do women employ in interactions with health personnel to secure support? Symbolic interaction was the theoretical foundation for the study, which included secondary analysis of 62 interviews with 20 women concerning their caregiving experience. In addition, new data were collected from 2 focus groups with 8 volunteers recruited from among the original 20 participants. The data indicated that the women's expectations of their caregiving role and their appraisal of the care recipient influenced their interactions with health personnel when seeking support. They employed 4 broad strategies: collaborating, getting along, twigging, and fighting/struggling. A woman's use of strategies varied according to the degree of mutuality in decision-making with staff and was accompanied by both positive and negative experiences. These findings confirm the importance of mutuality in relationships with health personnel and support the use of partnership and empowerment models of professional practice. PMID- 14746121 TI - A comparison of pain-assessment tools for use with elderly long-term-care residents. AB - The purpose of this study was to examine the psychometric properties (test-retest and interrater reliability, criterion concurrent validity) of 3 verbal pain assessment tools (Faces Pain Scale, Numerical Rating Scale, Present Pain Intensity Scale) and a behavioural pain-assessment scale for use with an elderly population. The study used a repeated-measures design to examine the reliability and validity of the tools across 4 groups of participants with varying levels of cognitive impairment using a non-random stratified sample of 130 elderly long term-care residents. The findings support the test-retest and interrater reliability of the behavioural pain-assessment tool across all levels of cognitive impairment, whereas the same measures of reliability for the verbal report tools decreased with increasing cognitive impairment; however, the majority of elderly with mild to moderate cognitive impairment were able to complete at least 1 of these tools. The findings are discussed in relation to their clinical and research implications. PMID- 14746122 TI - Development and psychometric evaluation of supportive leadership scales. AB - The purpose of this study was to develop and evaluate the psychometric properties of 2 supportive leadership scales, the Charge Nurse Support Scale and the Unit Manager Support Scale, designed for long-term-care environments. These 6-item self-report scales were administered to 70 nursing staff and their internal consistency reliability, test-retest reliability, content validity, factor structure, and construct validity investigated. Content validity was established with the assistance of experts. Both scales were deemed reliable. As hypothesized, a significant relationship was found between the measure of how nursing staff related to residents and measures of charge nurses' supportive behaviours (r = .42, p = .05). Reliable and valid measures of supportive leadership could be developed for use in identifying the quality of support provided to staff in long-term-care environments. PMID- 14746123 TI - Interventions for caregivers of persons with dementia: a systematic review. AB - The prevalence of dementia in older adults in increasing. Due to cuts in funding for formal support, many families are having to provide care for a loved one with dementia at home. This systematic review gathered and synthesized information on interventions designed to enhance the well-being of caregivers of people with dementia. A search of the literature resulted in 36 relevant studies. Of these, 11 were rated as strong, 11 as moderate, 13 as weak, and 1 as poor. This paper focuses on the strong studies. No one intervention had an overall significant impact on the well-being of caregivers. Although several interventions have been shown to be of benefit to caregivers, non-significant findings were more common. Further investigation is greatly needed. Systematic reviews are an important means of guiding consumers and practitioners in making evidence-based decisions. PMID- 14746124 TI - Recognizing and understanding the symptoms of type 2 diabetes. AB - Although diabetes education encourages people to monitor symptoms of glycemic imbalance, there has been little research on how people from ethnic minorities recognize and understand their symptoms. To explore this question, semi structured interviews were conducted with 23 African Americans over age 60 living with diabetes. Thematic analysis revealed 3 patterns of symptom experience. Those with prominent symptoms understood their body to be providing meaningful feedback on the diabetes and their efforts to manage the disease. Those with absent symptoms perceived no physical response to their diabetes, which they took to mean that the disease was well controlled. Those with perplexing symptoms could not interpret the messages they received from their body and were discouraged by this, often feeling they could not trust their body. The findings suggest that innovative strategies are needed to help people become more attuned to their body so they might better recognize and understand their symptoms. PMID- 14746125 TI - Factors in duration of anxiolytic, sedative, and hypnotic drug use in the elderly. AB - At least 2 decades of descriptive research on factors associated with psychotropic drug use by the elderly in the community has failed to yield convergent results. The authors posited that duration of use may have been confounding results of previous studies, since variables influencing initial use may not be those influencing long-term use. They conducted a secondary analysis of the elderly respondents in the cross-sectional 1998 Quebec Health Survey (n = 3,012). Results clearly show that factors associated with ASH use vary with duration of use. Apart from depression, medical and mental health factors significant for short-term use are not associated with long-term use. The only factors found that explain long-term but not short-term use were gender (female) and health perception (less than positive). These findings suggest that over the long term it is unlikely that mental health therapeutic benefits explain ASH use. The authors hypothesize that drug dependency could play a role in long-term use. They therefore encourage community health nurses to implement withdrawal programs in order to reduce harmful long-term consumption. PMID- 14746126 TI - Relationships between families and registered nurses in long-term-care facilities: a critical analysis. AB - Although much has been written about the relationship between families and nurses, little systematic analysis has been undertaken of this dyadic relationship in long-term care (LTC). Using a critical ethnographic approach, the researchers conducted separate in-depth interviews with 17 family-nurse dyads caring for residents with Alzheimer disease or a related disorder in one LTC setting. Analysis of interview transcripts and fieldnotes revealed 4 types of family-nurse relationships--conventional, competitive, collaborative, and "carative"--each reflecting the roles of nurse and family, negotiating strategies, and consequences. In addition, it became apparent that intrinsic and extrinsic factors influence the development of certain types of relationships. The findings have implications for nursing practice, policy development, and further research within LTC settings. PMID- 14746127 TI - Gerontological nursing research: a challenging but rewarding field. PMID- 14746128 TI - CIHR's Institute of Aging: improving the health and quality of life of older Canadians. PMID- 14746130 TI - Leisure-time activities--its program and importance in the institutionalized protection of old people. AB - This paper is a "report" or preliminary summation of a larger research project and paper. Leisure activities programs and their importance have not yet been systematically investigated in Croatian nursing homes, so this will contribute to a better understanding of this area of research. Through a ten year period of research study of 60 old people it has been shown that by the application of organized and suitable leisure activities we can prevent and redirect the measures so as to continually improve the life quality of old people living in nursing homes, regardless of their medical condition/place of residence. The topic of this paper is very popular in gerontological science. The research applied modern qualitative and quantitative methods of research in gerontology and therefore represents a novelty to the methodologically obsolete methods that have been in use in this country so far, which included polls and simplified quantitative processing of collected data. The results are useful for practical purposes because programs have been elaborated which will serve to improve the quality of leisure time and active life-planning in nursing homes. The foundations for further scientific research have been set with specific goals to focus on the certain aspects of the problems. In that sense, this paper invites all sorts of other challenging hypothesis to come out (e.g. the ratio of intellectual activities, active and passive types of activities etc.) and also opens the door for this kind of methodology in these types of research. This will help increase the number of such types of research as the qualitative methods of research have been disregarded in our country. PMID- 14746129 TI - A European project on health problems, mental disorders and cross-cultural aspects of developing effective rehabilitation procedures for refugee and immigrant youth. AB - The present paper describes the conceptual framework, rationale and methods of an international comparative study on risk and protective factors of adolescent health and well-being, with particular focus on youth with immigrant (or refugee) experience. This is a comprehensive study on the quality of life and health outcomes of adolescent youth that looks at group-specific differences within different socio-cultural contexts across six European countries, including those of post-conflict communities. The research project combines both quantitative and qualitative methods, using a common set-up across all countries involved with the goal of collecting data on 3,500 adolescents that are strictly comparable to allow cross-country analyses. It is particularly aimed at increasing the understanding of acculturation processes of a particularly sensitive population of adolescent refugees and immigrants and of the influence that the interaction of contextual and developmental factors has on their mental health and psychological well-being. PMID- 14746131 TI - Experiencing infertility--social work dilemmas in child adoption procedures. AB - The research deals with experiencing infertility and its consequences in the adoption of a child and focuses on infertile couples that have wished to adopt a child and joined a program preparing them to be foster parents. The results show that most of the infertile couples experience infertility very much as being different from couples with children as well as having to cope with the feelings of deep emotional loss resulting from the inability to reproduce biologically. There is therefore the question whether these facts should be taken into account by the profession (i.e. social workers) when dealing with child adoption as, according to most of the respondents of our survey, the process of coming to terms with infertility and its consequences is an important factor in establishing healthy family relationships and the child's identity within the adoptive family. We concluded from the results of the research that the infertile couples preparation program for adopting a child carried out by the Society of Adoptive Families "Deteljica", is a comprehensive autopoietic social workers' answer to the needs of participants for a successful adoption of a child, as it makes it possible to supply these future adoptive parents with the requisite information and experience and provides support to the entire family upon accepting a child in its midst, while its fundamental attribute is offering help to couples in overcoming the traumas resulting from their infertility. PMID- 14746132 TI - First toll free helpline for smoking cessation--analysis of results after one year of operation. AB - The aim of the study was to prove that there is a strong need among the smoking population for the antismoking telephone helpline, and to describe the structure and dynamics of calls to the Call-center. Basic data on socio-demographic characteristics, smoking habits, and reasons for calling the Center were collected during telephone conversations with smokers. The data were entered into previously prepared tables. Statistical analysis included 7,452 telephone calls; most calls were received from persons aged 26-45 years (34%), followed by 19-24 (24%) and 45-60 (19%) age groups. There was no statistically significant difference in the number of calls between men and women (54% vs. 46%, respectively). Most callers consumed on average 20 cigarettes per day. The most frequent reason for calling was to get on how to stop smoking, whereas seeking information on professional literature was the least frequent reason for calling among our respondents. Four-fifths of persons making a call to the Center started smoking when they were between 16 and 20 years of age. We can conclude that there was a need for this type of intervention due to its accessibility and potentially wide coverage of interested users. PMID- 14746133 TI - Acculturation process and its effects on dietary habits, nutritional behavior and body-image in adolescents. AB - Previous studies have indicated that the acculturation process may contribute to psychosocial and health problems among immigrants through the mediation of acculturation stress. This study focuses on adolescents with immigrant background permanently settled in the Croatian region of Dalmatia and the influences of the acculturation process on their dietary habits, nutritional behavior and perceived body-image. The survey was conducted on the total sample of 510 adolescents (aged 14-19) including 52 first generation and 248 second generation immigrants. The analysis included dietary habits and questions of restrictive diet as indicators of unhealthy influence on physical health. Psychological factors (stress and self esteem) have been found to be associated with dietary habits, diet behavior and dissatisfaction with body-image among adolescents with significant differences by immigrant status. Immigrant status is viewed as a risk factor for psychological distress and unhealthy dieting behavior. Research stresses the need to study the interaction between acculturation and health not only from an individual perspective, but also from the broader socio-ecological context of population subgroups. PMID- 14746134 TI - Comparative research on substance abuse and self-perception among adolescents with physical handicap. AB - The research on substance (alcohol, tobacco and drug) abuse and on self perception was done by comparing a test group of physically disabled adolescents and a test group of non-disabled adolescents. The respondents of the experimental group were students of the only special high school for physically handicapped persons in Croatia, Zagreb. The respondents of the control group were the students of two regular high schools in the capital of Croatia. The instrument used in this research was a self-reported, anonymous questionnaire. The respondents completed the questionnaire in the classroom. The data analysis regarding alcohol abuse indicated that physically disabled adolescents drink more often and out of quite different motives than their non-disabled peers. Regarding the prevalence, frequency, quantity and motives for smoking, no statistically significant difference has been found between the tested groups. On the contrary, significant differences between handicapped and non-disabled adolescents were evident regarding drug abuse. Only one physically disabled examinee used a drug- marijuana, only a few times a year. On the other hand, almost one quarter of the non-disabled adolescents use at least one, five at the most, type of drug sometimes or often. The results on the self-perception scale show that adolescent with physical disabilities have a much more negative attitude toward themselves than non-disabled controls. Their self-esteem and self-confidence are seriously diminished. Described findings could have a mighty impact on ways of preventing substance abuse, and on ways of increasing self-esteem among disabled and non disabled adolescents. PMID- 14746135 TI - Alu insertions in the Iberian Peninsula and north west Africa--genetic boundaries or melting pot? AB - The Western Mediterranean Basin joins a set of ethnically different populations as Iberians and Basques in the North shore and Berbers and Arab-speakers in the South one. In spite of this differentiation, they have maintained historical contacts since ancient times. The existence of a possible common genetic background (specially for Berbers and Iberians) together with the genetic impact of the Islamic occupation of the Iberian Peninsula during 7 centuries are some of the intriguing anthropological questions that have been studied in this area using several classical and DNA markers. The aim of this work is to present the results on a survey of polymorphic Alu elements in 10 human populations of the Western Mediterranean. Recent Alu subfamilies include a significant number of polymorphic Alu insertions in humans. The polymorphic Alu elements are neutral genetic markers of identical descent with known ancestral states. This fact turns Alu insertions into useful markers for the study of human population genetics. A total number of 14 Alu insertions were analyzed in 5 Iberian populations, 3 Berber groups from North-Western Africa, an Arab-speaker population from Morocco and a sub-Saharan ethnic group from Ivory Coast. The results of this study allow the genetic characterization of Berber populations, which show a certain degree of differentiation from Arab-speaking groups of the same geographic area. Furthermore, a closer genetic distance between South Spain and Moroccan Berbers as compared with other Spanish samples supports a major genetic influx consistent with some (but not all) previous genetic studies on populations from the two shores of the Gibraltar Straits. PMID- 14746136 TI - Genetic studies in south Balkan populations. AB - Within a study of the genetics of Balkan populations, four DNA-STR systems and 19 classical markers were examined in seven samples: Romanians (two groups), Albanians, Greeks and Aromuns (three groups). The results for the DNA-STR systems have been compared with data from the literature. The results show four clear separated groups: sub-Saharan black populations, North-African, Japanese and European populations. The large Balkan populations, except the Greek sample, are genetically more homogenous than the Aromun populations. A second Neighbor joining tree based on all 23 analyzed systems, show a particular trend of the Aromun groups, which indicates a particular genetic structure. PMID- 14746137 TI - Isonymy and the genetic structure of Albanian populations. AB - It is well known that in systems of surname transmission through the paternal line, surnames simulate neutral gene alleles belonging to the Y chromosome. This property of surnames was used to analyze the genetic structure of Albanian populations. Two large samples of surnames belonging to two different periods of time were analyzed. The analysis of indicators of population structure showed that geographical distance has an important effect on surname distribution. It seems that isolation by distance and genetic drift have been still important factors in the determination of the genetic structure of the Albanian population. PMID- 14746138 TI - Intergenerational trend of some dermatoglyphic traits in Vaidyas of West Bengal, India. AB - In order to investigate the intergenerational change of dermatoglyphics, fingerprints of 400 individuals were collected from an endogamous caste Vaidyas of Barasat, West Bengal. Results were compared with the data of an earlier sample of Banerjee collected in 35 years before on the same community of the same area. As it is generally known that dermatoglyphics is selectively neutral, thus if no other evolutionary forces play a role, we cannot expect any change of dermatoglyphic characters after several years. In the present study, non significant change in the frequency of pattern and more or less same PII have been observed in both sexes. But significant quantitative differences were found between the two samples. These differences may not be due to the change of intra uterine environment, rather due to the inter-observer error of these two studies and the small sample size of the earlier study. Because though same methods were used in both studies, inter-observer variation is much possible in ridge counting than pattern type determination. PMID- 14746139 TI - Molecular variation at functional genes and the history of human populations- data on candidate genes for cardiovascular risk in the Mediterranean. AB - A screening of 22 DNA polymorphisms has been performed in western Mediterranean populations (Iberian Peninsula, Morocco, and Central Mediterranean Islands). The analyzed markers correspond to polymorphic sites in several candidate genes for cardiovascular disease including apolipopoteins and their receptors (APOA1, APOB, APOE, APOC1, APOC2, LPA, and LDLR), genes implied in the hemostasis regulation (Factor VII, alpha and beta-fibrinogen, alpha and beta platelet-integrin, tissue plasminogen activator, and plasminogen activator inhibitor-1), and the angiotensin converting enzyme gene. The results are presented of a partial analysis carried out in following population samples: 6 from the Iberian Peninsula, 2 from Morocco, and 3 from Central Islands. The degree of inter population diversity was significant and consistent with data from other kind of genetic polymorphisms. The apportionment of the allele frequency variance supported a geographic structure into three main regions: Central Mediterranean Islands, the Iberia Peninsula and North Africa. The genetic distance pattern is compatible with a south-to-north North African influence in the Iberian Peninsula and a remarkable gene flow from sub-Saharan Africa into Morocco. Epidemiologically, North Africa is characterized by high frequencies of LPA PNR alleles with high number of repeats (protective for cardiovascular risk) and high frequencies of the APOE*E4 allele (risk factor) as compared with European populations. PMID- 14746140 TI - Insertion/deletion polymorphism of angiotensin-converting enzyme gene--risk factor for coronary artery disease in the Tuzla region population (Bosnia and Herzegovina). AB - Angiotensin II is the major effector molecule of renin-angiotensin system; its production can be conveniently interrupted by angiotensin-converting enzyme (ACE). Typical plasma levels of ACE accompany the I/D polymorphism; however, a controversy exists as to whether the DD genotype of the ACE polymorphism affects the risk for the development of coronary artery disease (CAD) and to what extent the ACE polymorphism is associated with CAD in different populations. We compared the I/D polymorphism in 212 CAD patients younger than 50 years with 165 healthy control individuals. They were all from the Tuzla region in Bosnia and Herzegovina. Patients with CAD had a higher prevalence of the DD genotype (36.3%) than controls (25.6%). The odds ratio for the ACE DD genotype in CAD patients was 1.7 (95% confidence interval 1.0-2.7; p < 0.05). We may conclude that the D/D genotype of the ACE gene polymorphism is associated with an increased risk for CAD in the Bosnian population. PMID- 14746141 TI - Cardiovascular risk factors in a small urban community in central Croatia. AB - The aim of this study was to determine in Karlovac (southern part of central Croatia) the most important risk factors for coronary heart diseases in men and women according to age < or = 59 and > or = 60 on the basis of their prevalence in 558 non-coronary patients and 442 symptomatic coronary patients. In younger male coronary patients (< or = 59 years of age) in relation to the control study, the statistically significant more frequent risk factors were hypercholesterolemia (p < 0.001), smoking (p < 0.01) and diabetes (p < 0.01). In older male patients (> or = 60 years of age) there was no statistically significant difference in a single risk factor. In younger female coronary patients, the statistically significant more frequent risk factors were hypercholesterolemia (p < 0.001) and diabetes (p < 0.001) and in older female patients diabetes (p < 0.05). This population sample showed higher prevalence of cardiovascular risk factors in younger coronary patients. The most frequent risk factors were diabetes, hypercholesterolemia and smoking. The difference is slighter in older coronary patients where it is diabetes, which is the most important for women. PMID- 14746142 TI - The incidence of stroke in Baranya county (east Croatia). AB - The aim of this retrospective study was to provide a survey of the incidence of stroke in Baranya, Croatia, on patients examined at Beli Manastir Health Center Department of Emergency from November 1, 1997 (the time of Baranya reintegration into the legal system of the Republic of Croatia after the war) till December 31, 2001. A total of 513 patients with symptoms of cerebrovascular diseases, or one patient every third day on an average, were examined. Total incidence of stroke was 16.09 per 10,000 population. The majority of patients were in the 61-80 age group with an incidence of 46.94/10,000 after the age of 60, 15-fold that was recorded in younger age groups. The most common risk factors recorded in examined group included hypertension, heart diseases, hyperlipidemia and diabetes mellitus. Total stroke mortality was 38.38%, whereas mortality in patients with hemorrhagic and ischemic stroke was 62.85% and 33.52%, respectively. The ratio of ischemic and hemorrhagic stroke in study subjects was 5:1, and in the causes of death 2.5:1. Out of 81 deceased stroke patients, 96.3% died within first 28 of admission. All of the patients with hemorrhagic stroke died within first 28 days, most within first 7 days (81.8%), whereas 94.9% of patients with ischemic stroke died within first 28 days. PMID- 14746143 TI - Circadian rhythm of blood leptin level in obese and non-obese people. AB - Leptin, an adipose tissue hormone, has circadian variations in its secretion. Aims of this study were to show how circadian rhythm depends on fat tissue distribution in obese and non-obese subjects. The research was carried out on 70 subjects (37 men and 33 women) with an average body mass index (BMI) of 25.22 kg/m2. Concentration of leptin in blood was measured at 8.30 a.m., 12.30 p.m. and 6.30 p.m. Basal leptin level correlated strongly with all isolated regions of subcutaneous fat tissue in women and obese subjects. Circadian changes of blood leptin level in non-obese people are more significant than these changes in obese people. Differences in circadian pattern of leptin secretion between obese and non-obese subjects were probably caused by enlarged volume of subcutaneous fat tissue in obese people. Lean subjects have subcutaneous fat in physiological range which allows influence of some hormones (insulin or cortizol) or food intake on leptin secretion. PMID- 14746144 TI - Development of centrality indices of subcutaneous fat during growth. AB - Changes of fat distribution were followed up in Czech and Slovak children from 1.5 to 15 years of age, using centrality indices, which relate the values of skinfolds on the trunk to the skinfolds on the extremities, head and neck. Up to 5 years of age, subcutaneous fat was deposited relatively more on the extremities, head and neck than on the trunk, which was expressed by lower values of the centrality indices. After the age of 5 years, the accumulation of subcutaneous fat was greater on the trunk, which was also expressed by higher values of the centrality indices. The comparison of the individual indices revealed in both genders a relatively higher amount of subcutaneous fat on the trunk in boys until 12 years of age. During puberty subcutaneous fat over triceps and on the forearm was reduced. In girls the deposition of the subcutaneous fat was relatively greater at different sites of the trunk than in boys, with the exception of the age of 14-15 years. The deposition of subcutaneous fat was greater on the trunk than on the head (cheek) and on the extremities in Czech compared to Slovak children, except for 12-year-old girls. During the period between the fifties and the seventies of the last century, in Czech children, especially in girls, the deposition of subcutaneous fat on the trunk was relatively smaller than on other parts of the body surface, which was expressed by the reduction of the centrality indices. Index 12 was therefore considered as the most valuable for the characterization of fat distribution on the body surface. PMID- 14746145 TI - Impact of war on growth patterns in school children in Croatia. AB - The objective of this paper is to present the growth patterns of school children in Osijek--the city which was exposed to severe attacks during the aggression on Croatia. The mean height and weight of Osijek schoolchildren aged 7 to 18 and the menarcheal age in girls in academic year 1995/96 were compared to the analogous data collected in 1980/81. The secular changes in height were heterogeneous. In older age groups from 12 in girls and 13 in boys, the mean height in 1995/96 increased markedly, whereas from 9 to 11 or 12, changes were undulating. In the youngest groups--at the age of 7 in both genders, and at 8 in boys, negative changes were observed. Markedly smaller height in this cohort was still pronounced in 1999/2000 when these children reached the age of 11. However, one year later (2000/01), at the age of 12, boys and girls caught up with their peers in the previous generations. These children during the war were approximately at the age of 2.5 to 4, a period when growth patterns are highly sensitive to adverse environmental influences. It might be possible that the emotional stress caused by a change of environment and separation from home, contributed to the deceleration of growth rate, i.e. the smaller height in a large part of childhood. PMID- 14746146 TI - Contribution to the physique of women with manic-depressive disorder in Hungary. AB - The purpose of this study was to get new data about the physique (somatotype) of manic-depressive patients. The somatotypes of manic-depressive females (n = 31, mean age: 30 year) investigated show a balanced mesomorphic-endomorphic predominance. The mean somatotype was 6.34, 5.27, 1.39. Previous Hungarian studies showed a meso-endomorphic somatotype in manic-depressive females. The physique of these patients determined by Kretschmer as pycnic did not show significant alteration due to environmental changes. Thus, according to the recent study, Kretschmer's statements (1921) are still valid in manic-depressive females. They are invariably characterized by a pycnic physique. PMID- 14746147 TI - Non-linear relations between selected anthropological predictors and psycho physiological exercise-responses. AB - In this paper, some very useful non-linear-relation procedures are actualized. The authors have defined the characteristic correlations between a set of anthropological characteristics (14 anthropometric and 14 motor-endurance status variables) and a set of psycho-physiological exercise-responses during the hi-lo and during the step aerobic dance training (heart rate, lactate concentration and rating of the perceived exertion). 60 healthy females served as the sample of subjects (mean age 21 +/- 1.4 years). The experiment consisted of two parts. In the first one, the linear correlations between the two sets of the variables were established. In the second part, non-linear (squared) relations, between the variables of the two sets were calculated. Results confirm the statement that the non-linear correlations, in some cases, better determinate the real nature of the relations between the variables, than linear correlation models. PMID- 14746148 TI - Phylogeny of vertebrate nuclear receptors--analysis of variance components in protein sequences. AB - Nuclear receptors (NR) constitute a large family of proteins and play a crucial role in regulating mineral metabolism and physiological homeostasis of various organ systems. The aim of this study was to elucidate whether the variance among NRs of estrogen, androgen and vitamin-D in various vertebrate species including humans is attributed to differences between the taxonomic groups within a specific receptor (i.e. between orthologous) or between the different proteins within the taxon (i.e. between paralogous genes). Published data on 57 protein sequences of the above NRs were used for phylogenetic analysis. The results showed that in DNA- and ligand-binding regions, 94% and 70% of variance is due to differences between the three proteins. However, in non-binding regions, 47% of the variance results from differences between the three paralogous proteins. Human sequences consistently clustered with their mammal orthologous within the three groups of NR sequences, clearly indicating that evolution of human sequences is not distinct from mammal sequence evolution. PMID- 14746149 TI - The fate of the arachnoid villi in humans. AB - Villi arachnoidales undergoes in the course of life changes in relation to the skull bones and sinuses. Our aim was to determine the relations of the villi arachnoidales to the skull bone and/or sinuses from the neonatal period to adults. The investigations were performed on collection of 50 disarticulated macerated skull bones from the new-born to 30 years of age and on 20 skulls from individuals in the life period from 30 to 80 years of age. Villi arachnoidales produced imprints on the skull bones in the shape of holes and/or furrows corresponding to different shape of the villi arachnoidales. These imprints appeared very early in the period when the bony sprouts of the large skull bones received a thin covering of compact bone, the future lamina vitrea. At that time villi arachnoidales had no connection with the dural sinuses but with the diploe and with the diploic veins. By agglomeration of the villi in larger and large formations, granula meningea, Pacchionian granulations, the contact to sinuses was realized by means of short channels. The structural changes of villi arachnoidales may produce thrombophlebitis and hydrocephalus externus, especially in children. The fate and the relations of the villi arachnoidales are therefore of great importance for neurologist, neurosurgeon and otorhinolaryngologist. PMID- 14746150 TI - Increase of classical antiepileptic drug utilization in Croatia during the process of introducing the new generation of drugs. AB - During the last decade the process of introducing the new generation of antiepileptic drugs (AEDs) has substantially changed the ways of treating epilepsy. Although a great deal of information about the role of new drugs has been accumulated, much less attention was paid to the impact of the new generation of AEDs on the utilization of classical AEDs. In order to detect the relation between the new and classical AEDs, the data about drug consumption in Croatia in the period 2000-2002 were analyzed. The main results indicated that the growth utilization rate (15%) was more the result of increasing consumption of the classical antiepileptic substances (in almost 2/3). It has been discussed that one of the possible interpretations for this phenomenon could lie in the fact that the continuing process of introducing the new AEDs was accompanied by a great number of educational activities. These activities have led to greater awareness of the facilities in treating epilepsy and consequently to a more active therapeutic approach, which encompassed both generations of drugs, even more the older one. PMID- 14746151 TI - Calculating lumbar puncture depth in children. AB - Lumbar puncture was performed in 195 children and the depth of needle was recorded. Our results show that the depth of lumbar puncture necessary to obtain uncontaminated cerebrospinal fluid correlates best with the child's weight. The simple formula: mean depth of insertion (cm) = 1.3 + 0.07 x body weight (kg), can be used to estimate the depth of lumbar puncture of children older than 3 months. The depths of lumbar puncture of children younger than 3 months are mostly 1.0 1.5 cm. PMID- 14746152 TI - The advancement in the early diagnostics of developmental hip dysplasia in infants--the role of ultrasound screening. AB - Developmental hip dysplasia (DDH) is a deformation of the locomotor system that can occur on previously normal bone structures in late fetal development. The study was undertaken to determine the value of continuous ultrasound screening in early diagnosis of DDH. In the area covered by the Labin Primary Health Center, clinical and ultrasound examination of the hips was performed in all children during the fourth month of life from 1989 to 2001. The incidence of DDH was 3.3%. The authors compared the obtained results with results of clinically examined hips in the period from 1968 to 1988 (incidence 1.7%), before the ultrasound diagnostics was introduced. The hip radiograms were performed only in children with clinically positive signs of higher degree of DHD. The ultrasound examination was performed with Hitachi EUB 410 real time linear transducer with a 5 MHz probe. The examiners used methods and criteria suggested by Graf. The most useful feature of the ultrasound method is that it allows the visualization of cartilaginous femoral head and its relationship to the acetabulum in infants. The method is non-invasive, repeatable, without risk of ionizing radiation or need for the contrast material administration. The results of the study revealed an important increase of DHD incidence after the introduction of ultrasound examination. US is highly sensitive in hip imaging and reveals the existence of a number of dislocated and subluxated hips that otherwise would be missed. The anomaly was discovered in the early period of life, which is very important for appropriate therapeutic effects. PMID- 14746153 TI - The structure of body measurements for the determination of shoe sizing for young Croatian men. AB - The determination and promotion of the system of shoe sizing requires accurate knowledge of morphological properties of the foot and lower leg of the tested population. Similarly it is necessary to establish the occurrence and regional distribution of definite sizes in the tested population. Possible regional differences in morphological properties must not be ignored because it has been established by means of anthropological measuring that both foot dimensions and foot shapes differ between populations and within the same population. This has been proven by the investigation of body measures, carried out for the purpose of establishing a system of footwear sizes, which was executed on a randomly selected sample of 4,268 healthy and normally developed males aged 18-22. The investigation was carried out in 1993 on five locations each of which representing a definite region of the Republic of Croatia: Jastrebarsko (central), Koprivnica (northwestern), Pula (southwestern), Sinj (southern) and Pozega (northeastern). The measuring instrument was 31 foot and lower-leg sizes according to the existing ISO standards for footwear. Interregional differences are significant in all body measures. The role and the contribution of individual measures to these differences has been established by means of discriminatory analysis with regions as a priori defined samples. In order to constitute a convenient and purposeful standard for the footwear size system it is necessary to work out a database with referent values for the same system. PMID- 14746154 TI - Holistic approach to functional anatomy of the injured ankle joint. AB - Talocrural joint injuries are among the most common injuries of the joints and therefore there is a need for a holistic approach to analysis of morphology, biomechanics and visualization of the talocrural joint ligamentary apparatus in different positions. The research was carried out on 20 fresh and conserved anatomical specimens of the lower leg on which X-ray, computed tomography, ultrasonography and stress analysis were performed before and after the lesion of ligaments. Also the gait of 130 adults without (100) and with ligament and joint capsule lesion (30) was analyzed by infrared telemetry. After complete discission of the lateral ligaments, arthrography and CT could register the lesion, while X ray and ultrasonography could not detect it. Gait analysis of healthy and injured leg showed that the injured leg was significantly less loaded. PMID- 14746155 TI - Orthotopic liver retransplantation--case report. AB - We report a case of orthotopic liver retransplantation (OLRT) in a patient who was suffering from rejection and graft failure after orthotopic liver transplantation (OLT). The patient was a 32-year old female who had diagnosed liver lesion--hepatic cirrhosis. Within two months, with presented condition as a terminal stage of her disease, she underwent the OLT and immunosuppressive postoperative management. Two months after the OLT, in the one-week period, the patient underwent two new operations because of obstructive icterus due to fulminant cholangitis and subhepatic abscess. In spite of this operative and conservative treatment the patient's condition did not improve. Because of graft failure due to bile duct necrosis, she underwent an ORLT operation and her condition is satisfactory, till now. We confirmed that the overall impact of retransplatation persists because patients undergoing elective retransplatation have significantly better prognosis than those requiring an emergency operation. PMID- 14746156 TI - A model of oncologic care in general medicine. AB - The problems related to cancer and its control initially manifest in local community, and general practitioners are those who most commonly have to face them there. The aim of the study was to develop a program of comprehensive oncologic care for primary care physicians, which would be highly professional, efficient, economically justified and feasible, with the ultimate goal of upgrading the target population health and quality of life. Opinions on the priorities and intensity of work in particular activities of general practitioners in the field of oncologic care were obtained from 54 Croatian experts in oncologic care. An Expert Opinion was designed to collect oncologists' opinions by use of Delphi method. The study was performed in two runs, yielding a high rate of accordance among the oncologists. 38 of 54 participants responded in the first run, and 40 of 54 (74%) responded in the second run. The results indicated pain therapy and terminal care to be given highest priority, whereas measures of primary prevention ranked first as a group. There was a unanimous agreement that current activities of primary care physicians in the field of oncologic care were not satisfactory, and that they should take the role of a coordinator of the oncologic care of both individual patients and the population at large. The study showed that a model of oncologic care applicable throughout the country could be developed by combining data from a small health care office with the knowledge of renowned experts in the field. PMID- 14746157 TI - The role of transbronchial lung biopsy in the diagnosis of solitary pulmonary nodule. AB - Transbronchial lung biopsy (TBLB) is a well-recognized diagnostic technique in diffuse interstitial lung diseases, but it is not considered to be the first choice in investigation of solitary pulmonary nodules (SPN). The main idea of this study was to increase the sensitivity of bronchoscopy using multiple techniques, especially TBLB, thus to avoid more aggressive diagnostic procedures. The objective of this prospective study was to evaluate the efficacy and safety of TBLB in the diagnosis of SPN, in comparison with other bronchoscopic techniques. Fifty patients with chest x-ray finding consistent with SPN underwent bronchoscopy with bronchial washing, brushing, bronchoalveolar lavage (BAL) and TBLB were included in this study. Thirty-one patients suffered from malignant tumors, while 19 patients had nonmalignant lesions. TBLB achieved overall diagnostic sensitivity of 62%, BAL of 29%, bronchial brushing of 16% and washing of 6%. Combining all techniques together, bronchoscopy had overall sensitivity of 86%. Concerning malignant lesions, TBLB had a sensitivity of 65%, specificity of 100%, and accuracy of 82%. TBLB had a significantly better yield for lesions with a diameter > or = 25 mm than for lesions of < 25 mm (sensitivity of 82% and 53% respectively, p < 0.05). Diagnostic yield improved significantly with the increasing number of specimens (less than 3 specimens: sensitivity 59%, 3 or more specimens: sensitivity 87%, p < 0.05). Complications of TBLB occurred in 2 (4%) patients: 1 incomplete pneumothorax and 1 hemorrhage. According to the results, we conclude that TBLB is an accurate and safe technique for the diagnosis of pulmonary solitary nodule with a diameter equal or greater than 25 mm. PMID- 14746158 TI - Early cancer in congenital choledochal cyst. AB - We report a case of 35-yr-old woman with early cancer in congenital choledochal cyst. She had a five-year history of intermittent right upper abdominal pain and intermittent jaundice. In this period she had a few abdominal ultrasonographies, but the cholelithiasis had not been found. Now, she was admitted to our hospital because she felt right upper abdominal pain with slight jaundice and subfebrile temperature four weeks ago. Abdominal ultrasonography showed enormous dilatation of the common bile duct, which was suspected as choledochal cyst. Computed tomography and endoscopic retrograde cholangiopancreatography revealed cystic dilatation of extrahepatic bile duct. An anomalous pancreaticobiliary junction was not found. The patient with congenital choledochal cyst was operated on. The excision of choledochal cyst was done with hepaticojejunostomy Roux-en-Y. There were no lymph nodes metastases. On the central part of choledochal cysts mucosa, it was shown a white plain area of thickness 0.3 cm and 0.8 cm in diameter. Histologically it was well-differentiated tubular adenocarcinoma, which was limited to the mucosa and which did not penetrate to other parts of the bile duct wall. From our knowledge, only small number cases of early cholangiocarcinoma in choledochal cyst were until now reported. Nearly eight years after the operation the patient feels very well, and has optimal working ability (Karnofsky 100%). PMID- 14746159 TI - Effects of war aggression in Croatia on some forms and manifestations of breast cancer. AB - The aim of this study was to determine the differences in epidemiological and clinical manifestations of breast cancer in time of war in Croatia and in peacetime before and after the war, in the defined population of Pozesko Slavonska County. The methods used in this study were the evaluation of relative predictive value of variables by means of chi 2-test and the analysis of variance, while the survival studies were tested by Long Rank test according to Kaplan-Meier analysis of survival. This work encompasses 660 patients who had breast cancer. The differences between three time periods were evaluated: the war period (1991-1995), and two control periods: before the war (1981-1990) and after the war (1996-2000). The patients were grouped by age, localization of tumor and survival. During the war period the patients were, on the average, 4.2 years younger than those who acquired the disease in control periods, and the mean age of patients was between 50 and 59 years (36.5% of patients). Although the difference in tumor distribution by sides (left or right breast) and quadrants was statistically significant (war period vs. control periods; p < 0.001), there was no statistically significant difference in the survival of patients according to the results of Kaplan-Meier analysis of localization of tumor (p > 0.05). The length of survival in terms of age of patients in time of surgical procedure was significantly different (p < 0.001). This study confirms the effects of war on some epidemiological and clinical manifestation of breast cancer in the defined population of Pozesko-Slavonska County. PMID- 14746160 TI - Unilateral multicentric breast cancer. AB - Clinical characteristics of unilateral multicentric breast cancer (UMBC) were explored depending on aggressiveness, survival rate, disease-free period and local recurrence. The study included 296 women with breast cancer, surgically treated between 1990 and 2001. UMBC was histologically proved in 29 (9.8%) patients. Multicentricity was defined by following criteria: a) tumor with minimum one satellite node in the same or other quadrant of the breast; b) minimum one cut through the breast without tumor cells; c) histopathologically, discontinued tumors with intra-ductal invasion. The average age of patients was 63.4 (range 36-85). There were 9 (31.0%) women with one satellite node, 7 (24.1%) women with two satellite nodes, and 13 (44.8%) women with three or more satellite nodes. At the operation, axilla was positive in 20 (68.9%) women. Steroid receptors were highly positive in 12 (41.4%) patients. Primary and secondary tumors were of the same histological type in 26 (89.6%) patients. Local recurrence was found in only 3 (10.3%) patients. A five-year period without disease was achieved in 24 (82.7%) women. Kaplan-Meier analysis showed a significantly higher survival rate at lower tumor stages (I or II) unlike in advanced stages with predominantly N2 grade. The results of this study showed a slightly lower five-year disease-free period than in the case of patients with monocentric breast cancer (MOBC). The survival rate was significantly lower at all advanced stages, especially determined by N2 axilla. Therefore, the conclusion is that multicentricity doesn't increase the risk of poor prognosis, especially at lower tumor stages. PMID- 14746161 TI - Effects of war aggression in Croatia on histopathological manifestations of breast cancer in defined population of one county. AB - The aim of the present study was to determine the differences in epidemiological and clinical manifestations of breast cancer during the war in Croatia and in peacetime. 660 consecutive patients were recorded (656 female and 4 male patients) from Pozesko-Slavonska County. The changes in histopathological features were recorded in war period (1991-1995, 156 patients) and through two control periods, before the war (1981-1990, 282 patients) and after the war (1995 2000, first five months, 223 patients). The relative predictive value was calculated using chi 2-test. The survival was calculated according to Kaplan Meier analysis of survival. The histopathological analysis showed an equal distribution of noninvasive cancer (in situ cancer) across periods. In the war period, the level of the most common invasive cancer, ductal breast cancer, was lower (57.7%), compared to control periods (71.2%:63.7%:68.2%). Opposite to that, invasive lobular cancer was more common in the war period (3.2%), compared to control periods (0.7%-1.3%). Furthermore, mixed cancer was also increased in the war period (7.1%) compared to control periods (0.7%-2.2%), as was medullar cancer (10.9% vs. 5.5%-5.9%). The study showed statistically significant differences in the survival of patients with different histopathological diagnoses (Log Rank = 47.49, df = 7, p < 0.0001), while the histological grade of tumor, as a predictive factor was not proved to be statistically significant (p > 0.05). This study confirmed the influence of war of war on histopathological incidence of some forms of breast cancer. PMID- 14746162 TI - Anthropological and clinical characteristics in adolescent women with dysmenorrhea. AB - The aim of this study was to examine the prevalence of dysmenorrhea in female adolescents and the influence of anthropological characteristics and lifestyle factors on menstrual pain. Two hundred and ninety seven girls from several elementary and secondary schools were interviewed about the presence of the menstrual pain, their age, height and weight, menarcheal age, menstrual cycles quality, smoking and sexual activity. There were 164 (55%) subjects with and one hundred and thirty three (45%) without dysmenorrhea. The adolescents with dysmenorrhea answered the questions about missing activities and taking pills for pain. No difference was observed between the girls with and the girls without dysmenorrhea in their chronological age, height, weight, menarcheal age, menstrual cycles quality, cigarette smoking and sexual activity. In the group of dysmenorrheic adolescents there was infrequent missing activities and bedrest, but missing school was observed in 22 percent and taking pills for pain was observed in 96 percent of the subjects. Young girls who experienced menstrual pain are good candidates for a prophylactic therapy, such as hormonal contraception. A replication of this study is needed for public health services in the future to improve the quality of life of the dysmenorrheic young women. PMID- 14746163 TI - Model of total skin electron treatment using the 'six-dual-field' technique. AB - During implementation of the total skin electron treatment, using six-dual-field technique, at radiotherapy department a large number of measurements are needed. To assess depth dose curve required by clinicians and dose uniformity over a whole treatment plane, combinations of different irradiation parameters are used (electron energy, beam angle, scatterers). Measurements for each combination must be performed. One possible way to reduce number of measurements is to model the treatment using the Monte Carlo simulation of electron transport. We made a simplified multiple-source Monte Carlo model of electron beam and tested it by comparing calculations and experimental results. Calculated data differs less than 5 percent from measurements in the treatment plane. During the treatment patient can be approximated using cylinders with different diameters and orientations. We tried to model the depth dose variations in the total skin electron treatment not just around the body cross-section (simplified to cylinders of different diameters), but also along the body to account for the variations in body curvature longitudinally. This effect comes down to the problem of modeling distribution in different cylinders, but varying the longitudinal orientation of those cylinders. We compared Monte Carlo calculations and film measurements of depth dose curves for two orientations of the cylindrical phantom, which were the simplest for experimental arrangement. Comparison of the results proved accuracy of the model and we used it to calculate depth dose curves for a number of other cylinder orientations. PMID- 14746164 TI - Changes in the interpupillary distance (IPD) with ages and its effect on the near convergence/distance (NC/D) ratio. AB - This study aims at determining the mean value of the ophthalmic anthropometrics parametar-IPD (Interpupillary distance) and the mean value of NC/D (Near convergence/distance) in 300 subjects aged 5 to 60 years. The influence of IPD on the NC/D ratio in the same subjects will also be investigated. The investigation showed that the mean value of IPD is 60.5 +/- 2.4 cm, and the mean value of NC/D ratio is 4.95 +/- 2.28 prD. Mean IPD of 5.1 cm in 5-year old children increases to 6.3 cm in adult over 20. In adulthood, IPD remains the same, i.e. 6.3 cm. Our study showed that the NC/D ratio is stable during lifetime. Since NC/D is stabile and IPD changes during lifetime, it is evident that factors other than IPD, such as convergence and accommodation, influence the stability of the NC/D ratio. PMID- 14746165 TI - A study of replacement of timolol-pilocarpine with latanoprost in pseudoexfoliation glaucoma. AB - The aim of the study was to evaluate the efficacy of replacing current dual local therapy (timolol and pilocarpine) with latanoprost 0.005% in 71 pseudoexfoliation glaucoma patients with controlled intraocular pressure (IOP). 39 patients switched to latanoprost 0.005%) and 32 patients continued timolol-pilocarpine therapy. Mean diurnal (IOP) was measured at baseline, after 0.5, 1, 3 and 6 months of treatment. After 6 months 38 patients with latanoprost and 30 patients with timolol-pilocarpine had completed the study. At baseline the mean diurnal IOP was 20.4 +/- 2.0 mmHg for patients in latanoprost treatment group and 21.4 +/ 2.1 mmHg for patients in timolol-pilocarpine group. At the end of the study, after 6 months of treatment, the mean diurnal IOP values were 16.6 +/- 2.4 and 17.9 +/- 2.0 mmHg respectively. IOP was statistically significantly reduced from baseline (p < 0.001). The mean diurnal IOP change from baseline was -3.3 +/- 0.5 mmHg (mean +/- SEM, ANCOVA) for the patients treated with latanoprost and -3.2 +/ 0.4 mmHg for the patients treated with timolol + pilocarpine. This difference in IOP reduction between groups was not statistically significant (z = 0.69; p = 0.49). This study showed that combination therapy (timolol plus pilocarpine) in pseudoexfoliation glaucoma can effectively be replaced by latanoprost monotherapy. PMID- 14746166 TI - Antioxidant enzymes activity in patients with peripheral vascular disease, with and without presence of diabetes mellitus. AB - The study evaluated antioxidant status in patients with peripheral vascular disease (PVD), with and without concomitant diabetes mellitus (DM). 211 participants were divided into standardized 4 groups: patients with PVD and DM (PVD+DM+), patients with PVD without DM (PVD+DM-), patients without PVD with DM (PVD-DM+) and patients without PVD and DM (PVD-DM-). The diagnosis of PVD was established by Doppler sonography analysis, including determination of the ankle brachial index (ABI), partial pressures along the leg, and CW Doppler sonography at typical locations. Antioxidant status has been evaluated through the colorimetrically assessed serum activity of key antioxidant enzymes: superoxide dismutase (SOD), catalase, and glutathione peroxidase (GLPX) as well as through total antioxidant status (TAS) determination. In PVD+DM- group, as well as PVD DM+ group, a significantly lower activity of the GLPX, catalase and TAS was found, whereas activity of SOD was significantly higher. There was no statistically significant difference between PVD+DM+ and PVD-DM+ group. Likewise, there was no statistically significant difference between PVD+DM- and PVD-DM group. This study has shown that there is statistically significant difference in activity of antioxidant enzymes between diabetic and non-diabetic patients, irrespectively of PVD presence. Furthermore, PVD present alone does not alter key antioxidant enzymes activity in comparison with healthy subjects. PMID- 14746167 TI - Pediatric urolithiasis in Croatia. AB - A retrospective review was performed of the records of 148 Croatian children with urolithiasis treated between 1989 and 2003. The study evaluated age, gender, family history, clinical symptoms, location of stone, laboratory findings, stone composition, mode of treatment and compared our results with data from higher and lower socio-economic countries. The mean age of our patients was 9.38 years (10 months to 18 years). Thirty-seven children (25%) were less than 5 years (group 1), 44 (29.7%) were between 5 and 10 years (group 2) and 67 (45.3%) were older than 10 years of age (group 3). There were 60 girls and 88 boys with overall male to female ratio of 1.47. Abdominal pain (83%) and haematuria (59.5%) were the main symptoms in the groups 2 and 3. Urinary tract infection was predominant symptom in the group 1 (62.1%). Calculi were located in the kidney in 90 children (60.8%), in the ureter in 39 (26.4%), in the bladder in 8 (5.4%). Urinary tract anomalies with or without infection were associate with a greater frequency of urolithiasis in the youngest age group and hypercalciuria was predominant cause in children over 5. Stone analysis was performed in 80 children. Predominant constituent of stones was calcium oxalate (48.7%), followed by struvite (25%), calcium phosphate (13.7%), cystine (10%) and uric acid (1.2%). Calcium oxalate stones were most common in all age groups. Struvite stones were most prevalent in the children younger than 5 years of age. Most patients (33.1%) underwent surgery for removal of their calculi. In 31.8% of children stones were passed spontaneously and the highest spontaneous passage rate was in the group 3 (37.3%). Stone composition, location and etiology in Croatian children are similar to those in developed Western countries. PMID- 14746168 TI - Craniofacial anthropometric pattern profile in hypohidrotic ectodermal dysplasia- application in detection of gene carriers. AB - Hypohidrotic ectodermal dysplasia (HED) is characterized by clinical manifestations of severe hypodontia or anodontia, hypotrichosis, hypohidrosis, and specific facial appearance. Affected males show complete expression of clinical features of this condition. Their mothers, who are gene carriers, express only some signs, which are usually very mild. Currently available clinical methods are not sufficient for routine identification of the HED heterozygous gene carriers. The purpose of this study was to identify and describe the facial characteristics of HED patients and their mothers and to evaluate the usefulness of craniofacial pattern profile analysis (CFPP) in the diagnosis of this syndrome and the detection of gene carriers. In this study six affected males and their mothers were evaluated. Z-scores for each variable were calculated and compared with age- and sex-matched controls. Anthropometric analysis showed a specific dysmorphic pattern in CST patients that includes decreased skull base width (t-t: -1.67 Z); decreased forehead width (ft-ft: -1.8 Z), decreased midface depth (sn-t: -2.02 Z), markedly decreased total facial height (n-gn: -3.4 Z), and markedly decreased maxillary arc (t-sn-t: -2.5 Z). Gene carriers showed a similar tendency in their pattern profiles. They showed the same tendency towards lower Z-values for forehead width, facial height, and mouth width. The values for these measurements were between those of the affected and healthy controls. The most pronounced findings were increased head width (eu eu: +2.83 Z), increased lower face width (go-go: +2.06 Z), and reduction of total facial height (n-gn: -0.95 Z). They also displayed increased nose width (al-al: +2.41 Z) and increased biocular distance (ex-ex: +2.01 Z). When used in conjunction with other methods the anthropometrics pattern profile analysis can considerably enhance detection of gene carriers for HED and increase objective assessment of the craniofacial region in HED patients. PMID- 14746169 TI - Occlusal molar surfaces in females with Turner's syndrome. AB - The aim of this study was to identify the molar occlusal features in 73 subjects with the Turner's syndrome (TS) and compared to a control group (CG) of 322 healthy females. The occlusal features were scored on dental plaster casts using the Scoring Procedures for Key Morphological Traits of the Permanent Dentition: The Arizona State University Dental Anthropology System (ASU). The results were analyzed through frequency, percentage and chi 2-test. TS subjects have more frequent reduction of cusp number, distolingual cusp on the upper molars and distal cusp on the lower molar, with the consequent reduction of the occlusal surface. Reduced size of occlusal surface and number cusps on upper molars resulted in the transformation of rhomboid occlusal shape into triangular, with the consequent loss of H-shaped groove system (in the upper right first molars H shaped groove system was significantly less frequently found in TS (p < 0.05); in the upper left second molars H-shaped groove system was significantly less frequently found in TS (p < 0.01). The X-chromosome aneuploidy can cause a decrease in developmental homeostasis, which results in the alteration of apposition of the enamel and in consequently substantial changes of the molar occlusal morphological features. PMID- 14746170 TI - Dental and minor physical anomalies in children with developmental disorders--a discriminant analysis. AB - A discriminant analysis was performed in a sample of 303 children with developmental disorders (DD) and 303 healthy controls (C) in order to test whether some oro-dental and physical minor anomalies could discriminate these groups of children. DD sample comprised 176 mentally retarded (MR) children. 70 children with impaired hearing (IH) and 57 children with impaired vision (IV). The control group included 303 healthy subjects, matched for sex and age. The analysis comprised seven common oral and dental anomalies: median diastema, hypodontia, impacted teeth, microdontia, dens invaginatus, upper lip frenulum and frenulum of the tongue. Minor physical anomalies were assessed by the method proposed by Waldrop et al., as the average number of minor anomalies per individual (W1) and as the weighted score of minor anomalies (W2). Three discriminant functions were obtained by analysis of nine initial variables. Distinct discrimination and considerable distances were found between the centroids of the controls and all groups of DD children. The first two discriminant functions were significant for discrimination between the groups and they explained 98.6% of the total variance. The first function contained 90.2% of information and was defined by the number and weighted scores of minor anomalies. The second variable explained 8.4% of the total variability and was defined by three dental anomalies. The results obtained by the discriminant analysis show that application of dental and minor physical anomalies enables discrimination between the group of healthy children and the groups of children with different developmental disorders. PMID- 14746171 TI - Patients' ranking of therapeutic factors in group analysis. AB - The aim of this research is to assess which therapeutic factors are of greatest importance to patients in group analytic psychotherapy, and whether the patients' characteristics and the phase of the group process influenced their evaluation of therapeutic factors. The Yalom's group therapeutic factors questionnaire was filled out by 66 patients, members of small groups conducted according to group analytic principles. The average scores for each therapeutic factor were subsequently ranked by importance to the patients and related to their age, sex, education, previous psychotherapeutic experience and phase of group process. Self understanding was the highest-ranking therapeutic factor for the patients (average score 21.32 +/- 0.04 out of 25 maximum), whereas identification was the lowest ranking factor (15.88 +/- 0.06 in average). Group therapeutic factors were scored higher by women, patients up to 30 years of age, high-school graduates, and those with previous psychotherapeutic experience. Self-understanding seems to be the most important therapeutic factor in group analysis, emphasizing the importance of appropriate selection of patients for group analysis in order to utilize therapeutic factors the best. PMID- 14746172 TI - A cephalometric comparison of skulls from different time periods--the Bronze Age, the 19th century and the present. AB - The aim of this study was to evaluate secular trends by means of orthodontic measurements on lateral cephalograms. We use roentgenograms from three populations: 22 Bronze Age skulls from a cemetery near Hainburg/Austria, 140 soldiers who served in the Hapsburg Imperial Army in the late 19th century, and 154 contemporary recruits of the Austrian Federal Army. Using conventional morphometric analysis, no statistically significant differences could be established. But applying geometric morphometrics to the 2D-coordinates of the pentagon composed of the landmarks Sella, Nasion, Articulare, Gonion and Menton, some biologically interpretable differences were detected, the size allometry between the 19th- and 20th-century populations being the only notable one. We conclude that landmarks should be digitized directly (and many more of them) and that conventional methods used in clinical orthodontics are inappropriate for addressing the scientific questions approached here. PMID- 14746173 TI - Anthropological analysis of the Late Roman/Early Medieval cemetery of Novigrad (Istria). AB - The paper presents results of analysis of human skeletal remains recovered from Late Roman/Early Medieval cemetery of Novigrad (Istria). The "terminus post quem" for the site was established archaeologically as 5th or 6th century A.D. The aim of this work was detailed bioarchaeological analysis of each individual. It included determination of sex, age at the time of death, reconstruction of body height, and detailed description of pathological changes on bones and joint surfaces acquired during lifetime. The analysis provides limited information on demography, health and disease of the ancient inhabitants of Novigrad due to the limited sample size. Results show unusually high proportion of subadults, a life span range of women slightly lower compared to other contemporary populations, a high level of metabolic stress in childhood and a high level of skeletal indicators of physical stress suggesting that several of the analyzed individuals were exposed to heavy physical labor during their adulthood. PMID- 14746174 TI - Empathy, communication, deception. AB - Empathy is understood as a mode of understanding operating on a subconscious level of mental processing. The cognitive element can only abstractly be distinguished from its affective expression. When recognizing a fellow creature we involuntarily sympathize with it. Recognition of covert motivations of overt behavior is the first step in formation of a communication channel between two (or more) empathizing agents. Yet, since communication evolved in variably complex social environments it was subject to pressure of conflicting individual interests. Deception thus evolved as an adaptive evolutionary strategy. Empathic understanding does not necessarily entail recognition of agent's real intentions. Deception may be achieved on both conscious and unconscious processing levels. A sufficient degree of biopsychosocial maturity must be reached for a child to be able to independently recognize verbal and non-verbal communication finesses. Once this level has been attained, the prevailing emotional orientation determines his/her degree of empathizing competence. PMID- 14746175 TI - Croatian-Hungarian radiological cooperation. PMID- 14746176 TI - [Applying deep lamellar keratoplasty in corneal disorders without endothelial abnormalities]. AB - PURPOSE: To present own experiences in the surgery of deep lamellar keratoplasty. MATERIALS AND METHODS: 25 eye of 25 patients suffering from various corneal pathologies with preserved endothelium: 22 with keratoconus eyes, 2 with post inflammatory scar and one eye with leucoma in Stevens-Johnson syndrome. Mean follow-up was 25.5 months (from 6 to 32). Main outcome measures were Non Corrected and Best Corrected Visual Acuity (NCVA, BCVA), astigmatism, refractive error, IOP and corneal transparency. The surgery was performed in general anesthesia. In majority of cases the recipient stroma was trephined to a depth of 70% with 7.5 mm diameter vacuum trephine. In cases of intraoperative perforation of Descemet membrane with shallow anterior chamber we converted the procedure into penetrating keratoplasty. The donor lenticule was trephined in the artificial chamber with the 0.5 mm oversized manual trephine to a depth of 90%. In two cases after DLK was finished, limbal stem cell grafting was additionally performed. RESULTS: Postoperative NCVA ranged from 0.01 to 0.8 and BCVA from 0.1 to 0.8. Astigmatism ranged from 0.5 to 10.0 D. Refractive error ranged from -8.0 to +1.0 D. In follow-up period all grafts maintained transparent. The complications were mainly intraoperative perforations of Descemet membrane (9 cases): in 7 cases we convert the procedure into penetrating keratoplasty. In 2 cases the double chamber-forming was observed: in one case on 7 day following surgery the penetrating keratoplasty was performed, in other a spontaneous attachment was observed. We also observed loose sutures in 2 cases, inflammatory infiltrates in one case. Two cases of ocular hypertension was successfully treated by medication. CONCLUSIONS: Deep lamellar keratoplasty is an effective procedure for treating various diseases of corneal stroma with unaffected endothelium. PMID- 14746177 TI - [Results of cataract extraction with IOL implantation in children aged 17 months to 8 years old]. AB - PURPOSE: To evaluate anatomical and functional results of cataract extraction and IOL implantation, together with posterior capsulectomy and anterior vitrectomy in children. MATERIALS AND METHODS: Cataract extraction and IOL implantation with posterior capsulectomy and anterior vitrectomy was performed in 32 eyes of children between 17 months and 8 years old. Patients were followed up for a mean period 36 months (from 6 months to 5.5 years). RESULTS: Good anatomical results were achieved in all, except 2 cases. One eye required removal of opacifications located posterior to the IOL In the second eye membrane from the anterior surface of the lens were excised. Final postoperative visual acuity in eyes with bilateral cataract was 0.59, whereas in unilateral cataract 0.25. Half of the children with bilateral cataract has significantly worse BCVA in one eye. CONCLUSIONS: Cataract extraction and IOL implantation with posterior capsulectomy and anterior vitrectomy in young children gives good anatomical results. Functional results are better in bilateral congenital cataract, comparing to unilateral cases. To achieve good functional results, visual rehabilitation is necessary. PMID- 14746178 TI - [Heparin-surface-modified PMMA intraocular lenses in children in early and late follow-up]. AB - PURPOSE: To compare anatomical and functional state of eyeballs after congenital cataract extraction and heparin-surface-modified PMMA (HSM-PMMA) or PMMA implanted lenses and to conclude, which type of implants is more advantageous in young patients. MATERIALS AND METHODS: 42 eyes of 30 children at the age of 3-16 years with congenital or developmental cataract after extraction and IOL implantation: I group--HSM-PMMA lenses in 24 eyes, and PMMA lenses in 18 eyes--II group. Mean age in the I group was 6 years and in the II group 8 years, p = 0.07. The presence of cellular and pigment deposits on the IOL surface as a sign of postoperative inflammation was examined by slit-lamp within 1 month after surgery. We have compared the results of visual acuity (V/A), corneal endothelium cell density and posterior capsule state. The examination were done in the short term follow-up mean 14.1 months (4-24 months) in the I group and mean 32.1 months (16-48 months) in the II group. In the long-term follow-up mean 38 months (30-44 months) in the I group, and mean 56.1 months (40-72 months) in the II group. RESULTS: Within first month after operation cellular and pigment deposits on the IOL surface were seen more often in the II group, p = 0.03. Best corrected V/A was comparable in both groups in the short-term and the long-term follow-up. At last visit, mean value of V/A was 0.54 (I group) and 0.42 (II group), p = 0.37. There was no significant difference between central corneal endothelial cell density in both groups: before (2893 +/- 261/mm2 and 2821 +/- 217/mm2, p = 0.143) and after operation (2371 +/- 202/mm2 and 2361 +/- 299/mm2, p = 0.428). In the short-term follow-up, but not longer than within first 12 months after surgery the frequency of clinically significant posterior capsule opacification (PCO) was less in the HSM-PMMA group (8%), than in the PMMA group (33%), p = 0.006. In the long-term follow-up there was no statistically significant difference in PCO appearance between these groups (44% and 50%, p = 0.27). CONCLUSIONS: Heparin surface-modified intraocular lenses reduced postoperative inflammation and delayed the incidence of PCO in children. Heparin-surface-modified intraocular lenses are more advantageous, than PMMA lenses in young patients. PMID- 14746179 TI - [The evaluation of accommodation range in patients of various age groups by WASCA wavefront aberrometer]. AB - PURPOSE: The aim of this study was, to analyse the results of accommodation range measurement obtained in groups of patients of different age. Wavefront aberrometer was used for evaluation of accommodation range of the patients eyes. MATERIAL AND METHODS: The accommodation range was analysed in 200 eyes in 100 patients. The examined group of patients was divided into 4 groups, depending on the patients age. GROUP I--AGE: 20-30 years old; group II--age: 31-40 years old; group III--age: 41-50 years old; group IV--age: 51-60 years old. The measurements were analysed using WASCA System depend on: refractive error, sex and difference of pupil diameter for far and near vision. RESULTS: Different ranges of accommodation were obtained. We did not observe any correlation between the accommodation range and the sex and refractive error. We noticed statistically significant differences between the accommodation range and pupil diameter, as well as the patients' age. CONCLUSIONS: The WASCA System is a device, which can be used for analysis of accommodation range. This system gives us the possibility, to trace changes of eye accommodation range in the long period of time. PMID- 14746180 TI - [Flicker perimetry (CFF) in glaucoma diagnosis]. AB - PURPOSE: Evaluation of CFF perimetry results in patients with suspected glaucoma. MATERIAL AND METHODS: Visual field was measured on Octopus 301 perimeter with option "flicker perimetry". 49 patients (98 eyes) with suspected glaucoma were tested. Standard W/W perimetry revealed glaucomatous visual field defect in one eye, but not in the second eye. Patients with cataract and diabetes were not taken into consideration. RESULTS: Defects in CFF perimetry tests were found in 71.4% of cases, but in W/W perimetry test in 50.0% of cases. CONCLUSIONS: Perimetry CFF is useful for early detection and follow up of glaucoma. Indices MD and LV may be used for those purposes. PMID- 14746181 TI - [Evaluations of ophthalmological care expectation in the opinion of countryside dwellers in the example of the Wola Uhruska region]. AB - PURPOSE: The aim of the study was the evaluations of ophthalmological care expectation in opinion of countryside inhabitants and their knowledge of eye diseases. MATERIALS AND METHODS: The analysis was performed based on the results of a survey, which covered 198 rural inhabitants living in Wola Uhruska region. The study was carried out during training arranged by Medical University of Lublin in July 2002. RESULTS: The obtained material was analysed and presented in the form of diagrams and written report. The results of the study showed, that 77% of patients was dissatisfied with the ophthalmologic health care access, despite, that the nearest ophthalmologic practice was in the distance of 30 km. Unfortunately, only 25% of persons could indicate the cause of their eye problems. It is important, to provide more information of eye condition for all patients. PMID- 14746182 TI - [Pars plana vitrectomy with corneal transplantation -- combined procedure]. AB - The aim of our study is to present own observations with Eckardt temporary keratoprosthesis, during combined pars plana vitrectomy and corneal transplantation. We operated on two aphakic patients with unclear corneas and retinal detachment--I case, phthisis bulbi after recurrent uveitis--II case. Eckardt temporary keratoprosthesis was sutured to the corneal bed with 4 or 6 Ethilon 10.0 bites, pars plana vitrectomy was performed followed by corneal transplantation and silicone oil tamponade. We obtained good transplant clarity only in the I case, in the II case because of hipotony and persistent contract of silicone oil with cornea, transplant was cloudy and collapsed with small exception in the central area. After 3-rd month we noticed local retinal detachment in the periphery, which was suppressed with laser photocoagulations. We think, that Eckardt temporary keratoprosthesis gives possibility to do vitrectomy in patients with undear cornea, which was in the past impossible. However, combined procedure requires surgical skills in both: anterior and posterior segments of the eye. PMID- 14746183 TI - [Acute posterior multifocal epitheliopathy: case report]. AB - PURPOSE: To present an atypical course of acute posterior multifocal placoid epitheliopathy. MATERIALS AND METHODS: 59 years-old diabetic man was seen at emergency because of a few days history of blurred vision in his right eye. Ophthalmologic examination revealed patches of blurred margine, multiple, yellowish lesions 1-2 dd in the fundus in both eyes. There were also lesions typical for non proliferative diabetic retinopathy like microaneurysms, dot and blot intraretinal hemorrhages and hard exudates. Optic discs and blood vessels were normal. We found peripheral drusen extending from the equator to the ora serata in both eyes. During 8 months of observation, we performed three times ophthalmologic examination, fluorescein and indocyanine green angiographies, USG A and B. No treatment was given. CONCLUSIONS: There is disagreement about the site of origin of placoid lesions, etiology and pathogenesis in acute posterior multifocal placoid epitheliopathy. In our report we agree with thesis, that the choriocapillaris are the primary site of the break. We propose, that different duration of the APMPPE and recurrences are directly proportional to the state and quality of the blood vessels. The possible explanation for longstanding lesions in our patient is diabetic damage of the capillaries in the retina and choroid. PMID- 14746185 TI - [Delayed massive suprachoroidal hemorrhage after retinal detachment surgery: case report]. AB - Massive suprachoroidal hemorrhage may occur at surgery, early postoperatively or may be delayed. The authors describe a case of 79-year-old woman, who developed a massive suprachoroidal hemorrhage in 72 hours after retinal detachment surgery. In our case, the main reason of suprachoroidal hemorrhage development was postoperative hypotony and occurrence of a lot of factors such hypertension, diabetes, high myopia, pseudophakia, which predispose to hemorrhage. This case is very rare but interesting because of its multifocal pathogenetical mechanism. PMID- 14746184 TI - [Vitrectomy in the treatment of endogenous fungal endophthalmitis in a patient after renal transplantation]. AB - The authors present a case of fungal endophthalmitis in the immunosuppressed patient after renal transplantation. The treatment comprised pars plana vitrectomy with silicone oil tamponade, peribulbar injections of Fungizone and intravenous Diflucan. Diagnostic and therapeutic difficulties in endogenous endophthalmitis are discussed. PMID- 14746186 TI - [Sequela after ocular trauma by injection needle]. AB - We present a case of patient with ophthalmological changes after ocular trauma by injection needle. PMID- 14746187 TI - [Changes in visual field of a child treatment with vigabatrin for 2 years]. AB - We described course of disease of a 12 year old boy with epilepsy, treated with vigabatrin. Pathological changes in visual field in both eyes are reported after 2 years of treatment. PMID- 14746188 TI - [Surgical techniques in limbal stem cell transplantation]. AB - Limbal stem cells (LSC) deficiency is one among the causes of corneal transparency loss. The only effective management is LSC transplantation. In available techniques the source of donor tissue containing LSC can be healthy fellow eye (CLAU, EVELAU), living-related (lr-CLAL, lr-EVELAL) or cadaveric donor (c-CLAL). From among them, the most efficient and the safest procedure is transplantation of cultured stem cells. Preoperative diagnosis and ocular surface condition determine the choice, of which procedure is the best. The surgical restoration of ocular surface is the sequence of procedures and the penetrating keratoplasty is the ultimate rehabilitative step in this process. PMID- 14746189 TI - [The role of vascular endothelial growth factor (VEGF)]. AB - The authors present the role of VEGF in microaneurysm formation, blood-retinal barrier breakdown, development of capillary nonperfusion and retinal neovascularization in pathogenesis of diabetic retinopathy. Inhibitors of VEGF in treatment of diabetic retinopathy are presented. PMID- 14746190 TI - [Treatment of retinal detachment of the course of retinopathy of prematurity]. AB - Cryo- and laser therapy of stage 3 have reduced, but not eliminated the occurrence of retinal detachment in stages 4a, 4b and 5 of ROP. In this disease the treatment of these stages is still the greatest challenge to the ophthalmologist. Therefore, the aim of this paper is to present our up-to-date possibilities of treatment of different kinds of retinal detachments in ROP, including segmental scleral buckling, encircling scleral buckling, scleral resection, vitrectomy and its modifications in ROP. Guidelines of surgery of retinal detachment in active stage 4 and 5 of retinopathy of prematurity have been described. PMID- 14746191 TI - [Influence of vigabatrin on visual fields and electrophysiological tests in patients with epilepsy]. AB - Vigabatrin (VGB) is a new antiepileptic drug, that acts by the irreversible inhibition of gamma-aminobutyric acid (GABA) aminotransferase. Chronic antiepileptic therapy with VGB is associated with visual field defects and electrophysiological changes. PMID- 14746192 TI - [Greek medicine in ancient times and its ophthalmological aspects]. AB - The article presents the state of medical knowledge in ancient Greece. Sacred medicine related to the cult of Asclepius and the origin of secular medicine is described. Ophthalmological aspects concerning the anatomy of the eye, diagnosis and treatment of ocular diseases are emphasized. PMID- 14746193 TI - [Daring to make a fresh start. Becoming a novice]. PMID- 14746194 TI - [Preoperative prevention and therapy of deep venous thromboses of the leg- current trends]. PMID- 14746195 TI - [Hormone therapy: treatment halt taking too long]. PMID- 14746196 TI - [Current studies reveal significant survival advantage. Breast cancer therapy with herceptin]. PMID- 14746197 TI - [Antimicrobial effect. Comparison of wound dressings containing silver]. PMID- 14746198 TI - [Fatigue--still an unsolved problem]. PMID- 14746199 TI - [Salt and stomach cancer: chances for healthy nutrition. Stomach cancer--what is really reliable?]. PMID- 14746200 TI - [Immunology. Bolstering the immune system with the power of fermented papaya]. PMID- 14746201 TI - [Legislation. Effective measures for preventing needle-stick injuries]. PMID- 14746202 TI - ["Up and down". Bipolar illnesses--underestimated prevalence and complexity]. PMID- 14746203 TI - [Mood stabilization as the goal. Expectations for ideal drug therapy of bipolar illnesses]. PMID- 14746204 TI - [Olanzapine (Zyprexa)--reliable in acute treatment, prevention of recurrence and long-term mood stabilization]. PMID- 14746205 TI - [18 million sick days due to depressive illnesses. New prospects in therapy of depression]. PMID- 14746206 TI - [Strategic cooperation of the Psychiatric Action Circle and the German Family Physician Association. Modern therapy standards in psychiatry]. PMID- 14746207 TI - ["Courage 2003". Politicians support the new school project "Crazy? So what!"- Benefit gala as for AIDS and cancer patients planned]. PMID- 14746208 TI - [Federal Family of Psychiatric Patients Association informs industry, pharmacies and medical practices about psychiatric illnesses. Family self-help psychiatry with a new educational campaign]. PMID- 14746209 TI - [Miele large capacity disinfection equipment facilitates flexible planning in central instrument sterilization department. New automated equipment: distribution into contaminated and sterile side even with little space]. PMID- 14746210 TI - [Nutrition therapy system--proper energy supply for every patient. General practice instructions for optimal fat supply in parenteral nutrition]. PMID- 14746211 TI - [New world-wide therapy standard determined by German Study Group. Decisive progress in ovarian carcinoma]. PMID- 14746212 TI - [Bone anabolic drug for severe osteoporosis. Forsteo available on the German market]. PMID- 14746213 TI - [2003 Responsible Care Contest]. PMID- 14746214 TI - [Does faith contribute to healing and health? "Your faith has helped you"]. PMID- 14746215 TI - Seeing the best and worst of humanity. Interview by Carl Bloice. PMID- 14746216 TI - [How surgery has changed and is still changing]. AB - Previous half of the century has influenced surgery not only by enormous progress of the knowledge in medicine, but namely in the last decade, by the introduction of mini-invasive operation techniques. Because of the highly effective conservation treatment, the spectrum of operations has been changed as well as the tactics and techniques of individual surgical procedures. Perspectives of surgery are in further specialization, introduction of one-day operations, and in the continuous development of mini-invasive operation techniques. The share of robots in the clinical praxis and in the training of surgeons is still difficult to forecast. PMID- 14746217 TI - [Heart transplantation in the Czech Republic and abroad]. AB - Paper presents brief history of the research in the field of heart transplantation in the Czech Republic and in the word. Indication criteria for the transplantation, operation technique, post operation treatment and long-term results are reviewed. It is considered that the heart transplantation became the routine clinical method, indicated to patients with irreversible heart failure when all other method of treatment had failed. The long-term survival is between 60 to 70%. PMID- 14746218 TI - [Lung transplantation--present status worldwide and in the Czech Republic]. AB - Lung transplantation has evolved to a standard treatment modality for patients suffering from end-stage lung diseases and it provides very good short- and satisfactory long-term survival. Accepted indications for lung transplantation include chronic obstructive pulmonary disease (COPD) and other causes of emphysema (i.e., alpha-1-antitrypsin deficiency), parenchymal diseases (i.e., idiopathic pulmonary fibrosis), genetic disorders such as cystic fibrosis, vascular diseases (i.e., primary pulmonary hypertension), chronic infectious diseases (i.e., bronchiectasis), as well as rare indications such as lymphangioleiomyomatosis or sarcoidosis. Presented article reviews the current strategies in the treatment of lung transplant recipients, surgical techniques, limitations of and effects of lung transplantation. PMID- 14746219 TI - [Liver transplantation--present status worldwide and in the Czech Republic]. AB - Since 1983 liver transplantation (LT) has been accepted as clinical method of treatment, namely of patients with "end stage liver disease", it is patients with the incurable phase of liver disease which excludes spontaneous recovery or successful conservation therapy, but which is not the terminal state. Patients, indicated for LT should be therefore in generally good condition, but with a prospect of survival with conservative treatment lower than one year. The most frequent indication in adults is the hepatic failure resulting from viral hepatitis and cholestatic diseases. In children younger than 2 years it is atresia of biliary ducts, in older children metabolic diseases and fulminant failure predominate. Children with secondary liver impairment resulting from a cystoid fibrosis or a hepatic malignancy are recommended for LT only rarely. About 8000 liver transplantations are performed annually in the world, about one sixth is in child recipients. Contrary to LT in adults, such operation in children has several risk moments. The liver splitting or grafting of a hepatic segment from a living relative donor belongs to the most complicated surgical techniques. At the end of nineties, in the advanced centers, one-year survival reached 85 to 90% with high probability of improving quality of life. In the Czech Republic 60 to 70 liver transplantations are performed annually in two centers (Prague, Brno) with comparable results. PMID- 14746220 TI - [Transplantation of the pancreas]. AB - Diabetes mellitus is a chronic disease with increasing prevalence in the population of the whole word. Transplantation of pancreas represents pro tempore the only treatment of Type I diabetes, which can assure long-lasting normal glycaemia and the independence from the insulin administration. However, transplantation still has higher morbidity and requires permanent immunosuppression. Authors describe their clinical experience with more than 200 transplantation of pancreas and they compare them with literary data. They review the basic problems and indications to the transplantation and in details describe the technique of the operation. Attention is given to the possible complications and to the immunological side of treatment. As a standard, the surgical technique has been using combined transplantation of pancreas and kidney. The most widely used method is the transplantation of the whole gland with a duodenal segment, which enables drainage of pancreatic juices into the intestine of the recipient. Due to the introduction of new immunosuppressives, it is possible to improve the long-term results of transplantations. Early diagnostics of the rejection of the pancreatic graft remains a specific problem. PMID- 14746221 TI - [Autologous hematopoietic cell transplantation in the Czech Republic]. AB - Transplantations of the autologous hematopoietic cell have been performed in the Czech Republic since 1993. Their total number has increased during the past ten years from 43 to more than three hundred procedures annually. Presented article describes, how the indications for autologous transplants have gradually changed according to the accumulating scientific evidence and it also gives examples of Czech researchers contributing to the advances in this field. Currently, autologous transplants belong to the standard treatment of a number of hemato oncological diseases and further studies, both national and international, are being performed in Czech Republic. Their aim is to improve this role of the treatment modality in the global care for patients with these diagnoses. PMID- 14746222 TI - [Allogenic bone marrow transplantation]. AB - Allogeneic bone marrow transplantation (alloBMT) represents an important part of the therapeutic approaches in many malignant as well as non-malignant hematological diseases. For some of them it represents the only curative strategy available, in others it may improve the outcome of treatment in high-risk stages. Curative potential of alloBMT consists of the curative potential of cytostatic activity of the pre-transplantation treatment and of so-called graft versus leukaemia effect (GVL), which is mediated by transplanted lymphocytes. GVL seems to be one of the most effective mechanisms in the treatment of many hematological malignancies and it is therefore studied in the context of the nonmyeloablative transplantations (minitransplantations). PMID- 14746223 TI - [Pure red cell aplasia in erythropoietin therapy in patients with kidney failure]. AB - Since the nineties of the previous century, incidence of pure red-cell aplasia (PRCA) in patients with chronic renal failure (CRF) and renal anaemia treated with recombinant human erythropoietin (rHuEPO) has significantly increased. Due to the positive effects of rHuEPO on quality of life, lowering of morbidity and mortality of patients with CRF, such increased incidence has attained a widespread interest, though PRCA remains only a rare complication. The responsibility for the development of PRCA lies with the neutralizing anti erythropoietin antibodies. The rise of antibodies and development of PRCA is related to the subcutaneous administration of erythropoietin and in the vast majority of patients to the treatment with Eprex, one of the epoetins alpha. At present, the most probable explanation is a change of the stabilizer in Eprex formulation, which is related to the increased immunogeneity of the product. The subcutaneous administration of rHuEPO, preferred for medical and economical reasons in both American and European guidelines, is known for its higher immunization power. Properties of the product, emphasized by the route of administration, can cause the rise of these antibodies. To prevent the rise of anti-erythropoietin antibodies and the development of PRCA, regulatory authorities and Eprex producers decided that Eprex cannot be administered to CRF patients subcutaneously, but only intravenously. Also the requirements on the handling of Eprex have become more stringent. Limitations do not concern either epoetin beta (NeoRecormon) or other epoetins alpha (of which the latter are not available in this country). Therapy of PRCA in patients treated with rHuEPO is based on suspension of rHuEPO and on the immunosuppressive therapy. Many questions concerning PRCA in CRF patients treated with rHuEPO remain unsolved. It is necessary to study further the ethiopathogenesis of this complication and possibly adjust preventive and therapeutic measures. PMID- 14746224 TI - [Tuberculosis in the Czech Republic in 2002]. AB - BACKGROUND: The number of all notified TB cases decreased in the year 2002. 1200 (11.8/100,000) cases of new TB diseases and relapses were reported in the year 2002. 972(9.6/100,000) were pulmonary TB and from them 658 (6.4/100,000) were confirmed pulmonary cases. METHODS AND RESULTS: The decreasing trend in TB incidence, reported since 1999, has continued. The treatment of confirmed pulmonary cases notified in 2001 and evaluated in 2002 was effective. Treatment success was in 71% of cases, but failures and interrupted were only 3.3%. Results of treatment were affected by many patients who died for other reason then TB, because they were from the oldest group of population. We have no influence on these deaths. The resistance for AT drugs was less then 7% and MDR cases were less the 2%. CONCLUSION: The low level of resistance and the effectiveness of treatment allow presuming a further decrease of TB cases. Next target of TB control should be active case finding in high-risk groups and monitoring latent infection. PMID- 14746225 TI - [Effect of fasting on gastric mucosa thickness: experimental study in laboratory rats]. AB - BACKGROUND: It has been proven that damage to the gastric mucosa is the result of the combined effects of acidopeptic agents including infection with Helicobacter pylori and insufficient protective mechanisms of the mucosa. Thickness of gastric mucus is very important protective factor. We can hypothetically expect that fasting may cause changes in the thickness of gastric mucus layer, which may in certain circumstances contribute to the conditions for mucosal injury. We have therefore conducted an experimental study on laboratory rats, aiming to assess the changes in the thickness of gastric mucus after 8 and 24-hour periods of fasting. METHODS AND RESULTS: We measured gastric mucus thickness with the aid of microelectrodes in 279 laboratory rats (group I) after 8 hours of fasting (2 measurements each, total of 558 measurements), followed by the same measurement in 70 laboratory rats (group II) after a 24-hour fasting. Group I had a gastric mucus thickness of 595 +/- 10 (mean +/- SD, microns); in group II the thickness was 502 +/- 10 (mean +/- SD, microns). After statistical analysis of the homogeneity of the two populations (Fisher's analysis), it has been proven that both groups belong to the same basic population sample. The statistical evaluation of the thickness of gastric mucus in both groups showed thinning of the mucus layer to 1% of the significant statistical difference. The difference between both groups is statistically significant. CONCLUSIONS: The thickness of gastric mucus is reduced after 24 hours' fasting in laboratory rats. This change is statistically significant (P < 0.01). It can be hypothesized that--under the specific conditions--this process can facilitate the injury of gastric mucosa. PMID- 14746226 TI - [Home care services yesterday, today and tomorrow in the Czech Republic]. AB - Paper presents description of the development of the home care system--since the experimental phase till to the projection of its outputs into valid legal standards of the Czech Republic. Authors discuss problems related to the standardization and stabilization of the integrated system of home care. Process of efficient induction of the home care by the general practitioners is described in relation to the specific indications and diagnoses. Multidisciplinary approach and interdisciplinary collaboration are required for granting of the home care of maximal quality and permanent availability for all indicated cases. Present state in Czech Republic is demonstrated on data sets recording development of the home care in the relation to effectivity and availability of the home care system for all citizens in the Czech Republic. PMID- 14746227 TI - [HIV/AIDS-positive patients and surgery. Part III. Surgical procedures in HIV positive patients and risk factors]. PMID- 14746228 TI - [HIV/AIDS-positive patients and surgery. Part IV. Case reports]. PMID- 14746229 TI - [Late manifestations of congenital diaphragmatic hernia]. AB - The aim of the work was to evaluate clinical and anatomical features in children with late manifestations of inborn diaphragma hernia after the newborn period. The group included 11 boys and 7 girls at the age of two to 78 months during the time of diagnosis. The diaphragma hernia was diagnosed in 15 children during non acute manifestations and in three children affected by emergencies. A posterior lateral defect of diaphragma was on the left side in 11 children and in 7 children on the left. A hernia sac was found in 11 children. Organs protruding into thorax included intestinal loops in 12 children, liver in 7 subjects, spleen in 6 individuals, stomach in five and kidney, pancreas and omentum in one each. The defect in diaphragma was occluded by a primary plasty in 17 children, while a patch from Goretex was used once. An intestinal strangulation required resection of necrotic part of intestine, but the boy died one month later for a multi-organ failure despite long-term resuscitation care. The other children were cured up without complications. The inborn diaphragma hernia should be considered in differential diagnosis of every child with unusual respiratory or gastrointestinal symptoms and abnormal X-ray picture of thorax. Acute strangulation of inborn diaphragma hernia after the newborn period is a life threatening disease, where the gastrointestinal obstruction is combined with respiratory and circulation failure. PMID- 14746230 TI - [Transanal resection of the recto-sigmoid--the future in the treatment of classic Hirschsprung's disease?]. AB - INTRODUCTION: The technique of transanal resection (TAR) of aganglionary part in rectosigmoid, published in 1998, has gradually become a standard operation technique in the treatment of Hirschsprung's disease (H.d.). Since the TAR technique for H.d. is not yet generally widespread and has not been used in the Czech Republic yet, the authors intended to share their own experience and early results obtained by this surgical technique. COHORT OF PATIENTS: Seven patients (five boys and two girls) suffering from the classical recto-sigmoid form have been operated on since June 2003. One girl was affected with the Down's syndrome. The age of the patients at the time of operation was between one and 24 months. SURGICAL TECHNIQUE: The extent of intestinal resection was determined on the basis of per-operation biopsy. The length of the rectrosigmoid resection was between 25 and 35 cm. The colorectal anastomosis with partial sphincteromyectomy of internal anal sphincter was performed by the technique according to Swenson. The antibiotic prophylaxis was secured by three doses of cefoxitin and one dose of isepamycin. RESULTS: Intestinal passage reestablished 8-12 hours after the surgery, and complete oral intake began on day 3 or 4 after the operation. All patients healed up without complications, the period of hospitalization was seven days on the average. The final diagnosis of H.d. was confirmed by biopsy examination of the dissected portion in all cases. CONCLUSION: TAR is univocally the method of first choice in the surgery of classical Hirsprung's disease when compared with laparotomy or laparoscopy. PMID- 14746231 TI - [Omentoplasty of the sternum--surgical solution in a sternal defect due to sternal osteomyelitis in a female patient with mediastinal localization of a parathyroid adenoma. Case report]. AB - We report here the case report of patient with empyema of the thorax and sternal osteomyelitis, which undertool removal of ectopic mediastinal parathyroid adenoma with omentoplasty of sternal defect. Osteomyelitis was cured; though empyema persisted for several months and it's healing was complicated with central venous catheter sepsis. PMID- 14746232 TI - [Complications after laparoscopic surgery of inguinal hernias]. AB - AIMS: The authors describe the occurrence of complications in laparoscopic approach to treat inguinal hernias in patients operated on at 2nd Surgical Clinic in Olomouc from May 1991 to the end of the year 2002. METHODS: The occurrence of preoperation and postoperation complications and relapses in inguinal hernias treated with laparoscopy in the period of May 1991 to the end of 2002. The TAPP method was used for operation on 98% of inguinal hernias, the IPOM method for 0.9% of inguinal hernias, TEP method in 0.4% and a simple suture in operation on 0.7% of inguinal hernias. RESULTS: Five hundred and thirty four (56% of all surgically treated inguinal hernias) were treated with laparoscopy in adult patients in the period of May 1991 to the end of 2002. Eight preoperation and postoperation complications (1.4%) were encountered. These included two cases of hematoma in the wound after port (0.3%), one hematoma of scrotum (0.2%), two artificial perforations of intestine (0.3%), one case of neuralgia of n. genitofemoralis, one artificial perforation of urinary bladder (0.2%), and one postoperation hydrocele (0.2%). There were 14 relapses (2.6%) in the 534 inguinal hernias operated on with laparoscopy. CONCLUSION: Based on our experience in the solution of inguinal hernias with laparoscopy the method of transabdominally preperitoneally localized grid (TAPP). In this method we have used three-point fixation--by a screw to pected ossis pubis and the upper margins of the grip with two transparietal stitches fixed permanently in subcutaneous tissue. PMID- 14746234 TI - [Magnetic resonance urography as an indicator for surgical treatment of anomalies of the upper urinary tract]. AB - AIMS: To compare the quality of MR of urography with intravenous urography, sonography and scintigraphy of urinary tract, presentation of MR urography as a possible substitution of so far used examinations in preoperation diagnostics of anomalies in upper urinary tract. METHODS: Static MR urography was used to examine 51 patients at the age of one month to 18 years with dilation of upper urinary tract detected by sonography. In 12 of them a dynamic MR study of the kidneys was also performed. Magnetic urography was compared with elimination urography, sonography and dynamic kidney scintigraphy. The detection rate was evaluated within the framework of the complex preoperation examinations for different stages of urinary tract in patients up to 2, 6, 12 and 18 years, respectively. RESULTS: In the total number of 36 kidneys, modified by hydronephrosis, magnetic urography was evaluated as more precise in 18 cases, elimination urography in eight and both examinations were considered equal in 10 cases. The imaging of megaureters by MR urography was more precise in 7 cases, elimination urography was at the same level six times and less precise in eight patients. The results differed in individual age categories. Sonography imaging of the kidney was good as well as for the lower portion of ureter but insufficient for the middle portion. The same course of excretion curves in dynamic excretion urography and dynamic scintigraphy of the kidney was reached in 6 patients. CONCLUSION: MR urography is capable to provide information on anatomy and function of the whole urinary tract in a single examination, being different from excretion urography by its ability of imaging and evaluating the hypofunctional part of the kidney. It represents a possibility to significantly reduce present examination which exerts a load upon the patient. PMID- 14746235 TI - [Benefits and risks of urologic laparoscopic surgery in adult patients]. AB - The paper describes the advantages and disadvantages of the laparoscopic operations, the number of which steadily rises in urology. The laparoscopic surgery is considered to be a benefit regarding the short postoperative hospital stay, painless postoperative course, and virtually non-existing postoperative paralytic ileus. As disadvantage are deemed the long learning curve for the operating personal, and high economical costs, which could be cut down only if short off-work period in productive population is included. In the paper, the pathophysiological guidelines are outlined and emphasized during the laparoscopic operation, which the surgical and anesthesiological teams have to have in mind. On the own cohort of patients, the numbers and types of operations are described, which have been done at our department. PMID- 14746233 TI - [CHS 100P surgical mesh in inguinal hernioplasty in adults using the Lichtenstein tension-free method]. AB - AIMS: The authors present their initial experience with surgical grid CHS 100 P with strengthened middle strip for the intestinal hernioplasty by the method without tension according to Lichtenstein. METHODS: In the inguinal hernioplasty by anterior transinguinal way in adult patients the authors use prolene grid and a surgical grid CHS 100 P with strengthened middle strip having been developed in collaboration with the Research Knitting Institute in Brno. RESULTS: From November 2001 to May 2003 56 adult patients were operated on inguinal hernia by the tensionless plasty according to Lichtenstein using a surgical grid CHS 100 P. No preoperation of postoperation complication or relapse were encountered. The patients were subject to load in the first postoperation day. CONCLUSION: Based on our first experience with flexible surgical grid with strengthened middle band CHS 100 P used for tensionless inguinal hernioplasty in adult patients according to Lichtenstein it may be concluded that the grid is well tolerated by the patients. Flexibility of the grid makes it possible to fully load inguinal area during the first postoperation day. PMID- 14746236 TI - [Recurrent prostatic carcinoma after curative radiotherapy and radical prostatectomy]. AB - INTRODUCTION: The authors evaluate indication criteria for individual therapeutic modalities of localized and locally advanced prostate cancer (CaP), but especially investigate progression of the disease in relation to CaP classification and other prognostic criteria. MATERIAL AND METHODS: The paper evaluates 120 patients altogether, treated by curative radiotherapy (RT) and 115 patients who underwent radical retropubic prostatectomy (RAPE). The patients were followed according to age, PSA value, Gleason score and life prognosis. The Partin nomograms were used to evaluate probability of localized disease, locally advanced disease, the involvement of seminal vesicles (gonecysts) and lymphatic nodes. The patients were also stratified according to T-classification, patients after RAPE also according to pT classification. RESULTS: RAPE: the mean period of observation was 37.9 month (6-114), 90 patients being without relapse (78.3%). A biochemical relapse occurred in 25 patients (21.7%). A local progression was confirmed by histology in 11 patients (44% of patients with progression, 9.6% of all patients after RAPE). RT: the mean period of observation was 26.75 months. A histological verification of the relapse was demonstrated in 14 patients (35.9% of patients with progression, 11.7% of all patients treated with RT). CONCLUSION: No progression was detected (under comparable period of observation) in patients with lower mean PSA values, a lower Gleason score and a higher probability of localized disease. In contrast, detection of a progression of the disease is significantly higher in patients with higher PSA values, a higher mean value of the Gleason score, higher probability of the locally advance disease or involvement of seminal vesicles. In order to reach good results in the therapy of prostate cancer for long periods of time, the disease must be detected in time and the stage of the disease should be determined exactly with a maximum precision possible. The prognosis in patients with worse "input" parameters is worse from the long-term point of view and these indices should be carefully controlled. PMID- 14746237 TI - [Multimodality therapy in progressive rectal carcinoma]. AB - Rectal carcinoma cannot be controlled only by surgical technique despite radical resection (TME, systematic lymphadenectomy). Therefore, progressive rectal carcinoma of stage 2 + 3 that are detectable pre-operatively by CT and 3-D endosonography, should be treated by a multimodal concept. Based on recent Scandinavian and Dutch studies, we introduced a modified post-operative short term radiation in which the acute toxicity on tumor tissue corresponds to that of the Swedish and Dutch studies (5 x 5 Gy), its late toxicity, however, is reduced by 20%. Thus, we expect to reduce long-term damages to the pelvic organs, i.e. incontinence. A phase 2 study revealed good therapeutic management due to interdisciplinary tumor conference and a high acceptance by patients. The post operative rate of wound healing is a bit higher, but does not influence the time of hospitalization. PMID- 14746238 TI - [Early immunomodulatory treatment of multiple sclerosis: pros and cons]. AB - Early immunomodulatory treatment of multiple sclerosis has raised great hopes, especially among MS patients and their families. However, there are considerable differences of opinion among medical professionals, particularly as regards the economic aspect of the therapy (often undertaken before any definite diagnosis is made). The findings documenting early irreversible axonal lesions and brain atrophy in MS, as well as results of clinical studies suggest that early immunomodulatory treatment is the method of choice, even if critical arguments are taken into account. PMID- 14746239 TI - [Brain metabolic disorders after chemotherapy in the study by magnetic resonance spectroscopy]. AB - The purpose of our study was to evaluate CNS pathology due to chemotherapy neurotoxicity, using MRI and localized proton MRS in patients with lung cancer treated with cisplatine, Vinca alkaloids and etoposide. A reduction in N acetylaspartate was expected as a result of chemotherapy neurotoxicity. METHODS: 31 patients aged 42 to 73 years underwent the following procedures before and after chemotherapy: clinical examination; MRI of the brain (Elscint Prestige 2T), MRS (PRESS sequence, TR 1500 ms, TE 80 ms) with volumes of interest (VOI) of 8 ml localized in the semi-oval center and a cerebellar hemisphere. The analysis of each patient's NAA/Cr and Cho/Cr ratios was carried out separately for the semi oval center and cerebellum measurements. RESULTS: None of the patients demonstrated any clinical manifestations of the CNS neuropathy. MRI of the brain did not reveal any abnormalities caused by chemotherapy. Pre-treatment NAA/Cr and Cho/Cr ratios in the semi-oval center did not differ significantly from these measured after chemotherapy. However, the analysis of the cerebellar spectra showed a significant decrease in the NAA/Cr ratio (p < 0.05) and a time-related decrease in the Cho/Cr ratio (p < 0.05) after chemotherapy. An analysis of Pearson's correlations showed a very strong linear relationship between NAA/Cr and Cho/Cr ratios (p < 0.001), both in the semi-oval center and cerebellum. CONCLUSION: The decreased NAA/Cr ratio can indicate some neuronal loss caused by chemotherapy. The decrease in the Cho/Cr ratio could be associated with some myelin damage. The MRS results suggest the presence of a sub-clinical selective cerebellar neuropathy caused by chemotherapy. The MRS revealed that reaction to chemotherapy was different at the semi-oval center than that in the cerebellum. The results allow theorizing about an alternative or two-stage brain response to the neurotoxic factor found both in the cerebrum (the semi-oval center) and cerebellum. These initial results indicate that proton MR spectroscopy is a potentially useful modality for detecting an early stage of the CNS pathology caused by neurotoxicity of chemotherapy. PMID- 14746240 TI - [The assessment of development of binding antibodies to interferon beta during interferon beta 1a treatment of multiple sclerosis]. AB - The importance of binding antibodies (BAb) that develop during the treatment of multiple sclerosis with interferon beta has not been fully explained yet. However, they are generally regarded as one of factors that may diminish treatment efficacy. The aim of the study was to evaluate firstly, BAb occurrence in interferon beta 1-a (IFN beta 1-a)-treated MS patients and secondly, BAb impact on clinical efficacy of this medication. In the 36-month study participants were 21 patients with relapsing-remitting multiple sclerosis, RR-MS, (14 women, 7 men, aged 29.6 +/- 8.5). All the patients were receiving intramuscular IFN beta-1a (Avonex) for 24 months, in the dose of 30 micrograms per week. Clinical parameters and serum BAb levels (the EIA method) were estimated every 3 months. Two control groups, examined only once, included 20 RR MS patients without IFN-beta therapy and 20 healthy volunteers. While before treatment a high BAb level was found in 2 patients (9.5%), at 6 months of treatment it was found in 8 patients (38.1%). A similar number of patients with high BAb levels was seen throughout the study during the IFN-beta treatment. On therapy completion serum BAb levels decreased very rapidly. After 2 years of treatment, disability as measured by the EDSS scale was more pronounced in patients with serum BAb, but the differences were statistically not significant. No statistically significant relationship was found either between elevated BAb levels and the number of relapses during the IFN-beta treatment (including relapses that required steroid therapy). PMID- 14746241 TI - [Diagnosis and treatment of carcinoid meningitis: a challenge to the neurologist and oncologist]. AB - BACKGROUND: Two clinical types of leptomeningeal metastases from solid tumors are observed: local and disseminated. The former (meningeal carcinomatosis) consists in nodular infiltration of leptomeninges, while the latter (carcinomatous meningitis)--in tumor cells free floating in the cerebrospinal fluid and adhering as a monolayer to the surface of neural structures. Despite the same etiology, the two types of metastasis differ in their clinical manifestation and prognosis. Meningeal carcinomatosis is more frequently diagnosed nowadays due to advances in neuroimaging techniques and the long survival of breast cancer patients. Patients with local, nodular infiltration of leptomeninges may survive many years without symptoms of the disease. On the other hand, carcinomatous meningitis, with its usually violent course and short survival, has become a major problem for oncologists and neurologists because of limited efficacy and considerable toxicity of the treatment. AIMS: The purpose of this article is to review the current knowledge about carcinomatous meningitis in breast cancer patients, taking into account pathophysiology, clinical symptoms, diagnosis, treatment and prognosis. The second aim was to present the authors' experience with the treatment of breast cancer patients suffering from carcinomatous meningitis. MATERIAL AND METHODS: 37 patients with breast cancer and carcinomatous meningitis were treated in the Oncology Center, Warsaw, in the years 2000-2002. Their mean age was 51. The diagnosis was based on results of neurological examination, MRI scan, and the presence of neoplastic cells in the cerebrospinal fluid. In a majority of cases combined treatment was applied, including intrathecal administration of cytostatics, intravenous systemic chemotherapy and radiotherapy. RESULTS: The observation period ranged from 2 to 33 months. A response to the treatment was achieved in 76% of the patients. Their median overall survival was 19 weeks, mean 18 weeks. Seven patients (19%) survived for over 6 months. CONCLUSIONS: The ever-growing incidence of carcinomatous meningitis in the course of breast cancer has become a serious clinical problem for neurologists and oncologists. Treatment results are disappointing, although the combined modality treatment appears to be the best option. New pharmacological approaches to the treatment of meningeal malignancy are required to improve the outcome of patients with carcinomatous meningitis. PMID- 14746242 TI - [Changes of cerebral blood flow in the basilar artery system in patients with neurogenic syncope]. AB - Syncope is a short loss of consciousness caused by a transient decrease of global cerebral blood flow. Neurally mediated syncope is associated with vasodilatory response and bradycardia. The Doppler ultrasonography was used to evaluate changes of the basilar artery blood flow mean velocity and pulsatility index during orthostatic challenge and CO2 reactivity testing in patients with neurally mediated syncope and a negative result of the Tilt Test. Eighteen patients aged 20-40 years with a history of cerebral syncope and 10 healthy volunteers were qualified. RESULTS: There were no significant differences between syncope patients and controls in the basilar artery blood flow parameters in various body positions during normocapnia, hypocapnia and hypercapnia. Irrespective of body position, the reactivity index in patients with neurally mediated syncope was lower than that in controls, but not significantly. CONCLUSIONS: No significant pathology was found in cerebrovasoreactivity responses in the area supplied by the basilar artery, which suggests that cerebral autoregulation disorders cannot be the primary cause of neurally mediated syncope. PMID- 14746243 TI - [Transcranial color Doppler sonography (TCCD) in the diagnosis of vasospasm in the middle cerebral artery. Evaluation of TCCD diagnostic effectiveness by means of receiver operating characteristics curve (ROC) analysis]. AB - Transcranial Doppler sonography is commonly used for diagnosis of cerebral vasospasm. However, the overall diagnostic performance of this method in detection of arterial narrowing has not been established. Blood velocity threshold, diagnostic for vasospasm, has been proposed for conventional, "blind" transcranial Doppler sonography (TCD). Nevertheless it cannot be used for transcranial color Doppler sonography (TCCD), because in this method the obtained blood velocities in the major cerebral arteries are higher than are in TCD. The aim of this study was to estimate the accuracy of transcranial color Doppler sonography in detection of middle cerebral artery (MCA) narrowing by means of receiver operating characteristic curve analysis (ROC). One hundred thirty four patients were studied with TCCD immediately before cerebral angiography. There were 75 men and 59 women, age range from 18 to 74 years, mean age 49 years. Of the 268 MCAs examined, 227 arteries were finally included in the construction of the ROC curve. Angiographic vasospasm was graded as none, mild (equal to or less than 25% of vessel caliber reduction--16 patients) and moderate-to-severe (more than 25% of vessel caliber reduction--29 patients). The overall diagnostic performance of transcranial color Doppler sonography in the detection of moderate to-severe vasospasm of middle cerebral artery was found to be very high. The value of associated area under the ROC curve was 0.94. The value of this area for all vasospasms was 0.85, and this indicates good accuracy of a test. The best performing TCCD parameter for the detection of MCA narrowing was revealed to be peak-systolic velocity. The optimal trade-off between sensitivity and specificity was achieved with a peak systolic velocity of 182 cm/s. Therefore this velocity threshold is proposed as most universal for the diagnosis of vasospasm in the middle cerebral artery. PMID- 14746244 TI - [Direct trans-sylvian approach to the ventrolateral pons in surgical management of large cystic cavernous malformations of the brain stem in children]. AB - OBJECTIVES: In the study surgical indications are reviewed and the effect of the applied surgical technique on long-term treatment outcome in children with large cystic brainstem cavernous malformations is evaluated. METHOD: Clinical data of 5 patients treated surgically for large cystic cavernous malformations of the pons in the years 1995-2001 were retrospectively evaluated. In all the cases the transsylvian approach to the ventrolateral pons with splitting of the crus cerebri was used to reach cavernoma within the cystic cavity. The long-term follow up (mean 4.9 years) included a complete neurological examination and annual MRI studies. RESULTS: At least one hemorrhagic episode was diagnosed in all the cases, while bleeding to the pons, with clinical course mimicking that of stroke, was noted twice in 3 patients. All the episodes confirmed by MRI imaging appeared to be intralesional. The malformations ranged in size from 30 to 50 mm (mean 36.9 mm). Their most characteristic MRI features were not only the localization in the pontine structure exclusively i.e. within the natural anatomical barriers separating the pons from the midbrain and medulla, but also the growth pattern corresponding to that of brainstem benign gliomas. The patients' mean preoperative KPS score was 60 (30 to 90). Cavernous hemangiomas were totally removed in 3 out of 5 cases. One patient with partially removed lesion presented after a year recurrent hemorrhage and new neurological deficits that required a second stage surgery. In all but one case transient complications included some new cranial nerve, motor and sensory deficits that influenced the patients' immediate postoperative KPS assessment, with the mean score of 48. In a follow-up assessment at 1.5 to 7 years, a significant amelioration of both their neurological status and quality of life was found, with KPS scores from 70 to 90 (mean score 80). Annual MRI examinations showed extensive hemosiderin deposits within the pontine tissue in 3 cases of totally removed cavernous hemangiomas and this image was stable during the follow-up period. CONCLUSIONS: The findings suggest that resection of large pontine cavernomas in children is indicated in cases of symptomatic hemorrhage with lesions approaching the pial surface, or surrounded by a small margin of normal tissue. Recurrent hemorrhages are intralesional (i.e. limited to pontine structures by the natural anatomical barriers), resulting in a "cyst-like" growth of malformations mimicking that of focal neoplasms and in compression of the brain stem tissue rather than in a direct hemorrhagic insult. PMID- 14746245 TI - [Endovascular embolization of cerebral arteriovenous malformation (AVM)]. AB - The paper presents the technique and outcomes of brain arteriovenous malformation (AVM) endovascular treatment using a glue-lipiodol mixture. Embolization was attempted in 33 AVM patients (18 women). In 6 cases (18%) AVMs were complicated by brain aneurysms. In 20 patients (61% of cases) the presence of AVM was manifested by intracranial bleeding. RESULTS: 87 procedures were performed; the nidus was completely embolized in every case, total occlusion of AVM was obtained in 12 (35%) cases, subtotal (i.e. 75%) in 13 (36%) cases, and 50% occlusion--in 9 (28%) cases. Out of 8 (24%) cases in which complications occurred, one patient (2%) died. PMID- 14746246 TI - [Hyperhomocysteinemia in patients with dementia]. AB - Angiogenic brain damage and Alzheimer's disease caused by a progressing degenerative process are listed among the most frequent causes of dementia. These two processes are often concurrent and interrelated. Risk factors for vascular diseases including hypercholesteremia, arterial hypertension, and diabetes are also recognized risk factors for Alzheimer's disease. Results of many studies conducted in recent years suggest that the atheromatous process may be induced by elevated levels of homocysteine. Hyperhomocysteinemia first and foremost accelerates the onset of microangiopathic changes in small vessels. The mechanism underlying atherogenic action of homocysteine is still unclear. Hyperhomocysteinemia, generally assumed to have cytotoxic properties, damages endothelium in blood vessels, enhances thrombotic changes, and directly acts upon nitrogen oxide (NO), a vessel-dilating factor. Homocysteine is a metabolite of methionine. Homocysteine metabolism depends on current needs of the organism and involves either methionine reproduction (the reaction of remethylation, with such cofactors as B12 vitamin and folic acid), or cysteine synthesis (the transsulphuration reaction, with B6 vitamin as a cofactor). The normal range of plasma homocysteine concentration is assumed to be 5-14 mumol/L. The prevalence rates of hyperhomocysteinemia are 3-7% in the general population and 25% among those with vascular diseases. Elevated plasma homocysteine concentrations are due both to genetic and to environmental factors. In 2/3 of cases hyperhomocysteinemia is caused by decreased levels of folic acid, pyridoxine, and cobalamin. Deficiency of these vitamins is often seen in healthy elderly people. PMID- 14746247 TI - [Relationship between stroke and hormonal changes in various periods of life time]. AB - Strokes are still considered a major social problem. The purpose of this paper is to discuss hormonal changes in various periods of life in the context of stroke occurrence. The influence of menopause and andropause on cerebrovascular diseases is described and the association between such conditions and osteoporosis is analyzed. Particular attention is paid to the incidence of strokes in pregnancy, as well as among women receiving oral contraceptives and replacement therapy. PMID- 14746248 TI - [Subsidence and its effect on the anterior plate stabilization in the course of cervical spondylodesis. Part I: definition and review of literature]. AB - A definition of subsidence in terms of spinal biomechanics is presented in the paper. Subsidence is defined as sinking of a body with a higher elasticity modulus (e.g. graft, cage, spacer) in a body characterized by a lower elasticity modulus (e.g. vertebral body), resulting in 3D changes of the spinal geometry. Magnitude of subsidence is directly proportional to the load pressure and to the difference between the elasticity modules, but inversely proportional to the area of the graft-bed interface. Both biological and mechanical qualities of the graft bed interface are important for the subsidence process. Any excessive subsidence decreases the interbody space and produces both local and general kyphotization of the spine. This may cause destabilization of the screw-plate and/or screw-bone interfaces (e.g. pulling-out, altered angulation or breakage of the screws). A method is proposed of radiological estimating the absolute magnitude of subsidence, based on the real known length of the implanted stabilizer (e.g. plate). Clinical examples of an excessive subsidence and its impact on the stabilizing plate system are presented. Subsidence is inherent in the interbody fusion process. Endplate preservation and a dynamic modification of cervical plates may enables us to control subsidence and reduce the number of complications. PMID- 14746249 TI - [Fronto-temporal dementia: case report]. AB - Frontotemporal dementia is a relatively common form of primary dementia. The article presents a case of a female patient with probably inherited frontotemporal dementia, originally diagnosed as Alzheimer's disease. PMID- 14746250 TI - [Removal of multiple intracranial meningiomas preceded by intravascular embolization in a Jehovah's witness patient. Case report]. AB - The paper presents a case of a Jehovah's Witness, 64-year-old woman harboring multiple meningiomas in the right cerebral hemisphere. As the first stage of her treatment endovascular embolization of the tumor vascular bed was performed. Through the middle cerebral artery Ivalon particles and acrylic glue were injected. Four days later the patient underwent craniotomy and four intracranial meningiomas were removed. Intraoperative blood loss was minimal and the patient was discharged with no neurological sequelae and without anemia. The authors emphasize benefits that endovascular embolization may bring, especially as regards improvement of neurosurgical treatment safety in Jehovah's Witnesses. PMID- 14746251 TI - [Post-traumatic unilateral atlanto-axial rotatory subluxation in an adult]. AB - This retrospective single-patient case report deals with a rare form of spinal trauma, i.e. atlantoaxial rotatory subluxation. The authors present a review of the literature including a classification of rotatory atlantoaxial subluxation types proposed by Fielding, and describe their own experience with treatment of this condition. A case is reported of a 29-year-old woman with a history of head contusion in a car accident. Immediately after the trauma she had the following symptoms: torticollis and neck pain with decreased cervical motion. Atlantoaxial rotatory subluxation of the right C1-C2 articulation was diagnosed by plain radiographs and CT. The patient was treated surgically by an open reduction, unilateral screw fixation of the right CI-C2 articulation (according to Magerl) and posterior C1-C2 wiring with graft. The normal atlantoaxial relation was restored with disappearance of torticollis. At follow-up certain limitation of neck movements due to C1-C2 stabilization persisted. No complications were noted. The patient remains neurologically intact and has radiographic documentation of fusion. PMID- 14746252 TI - [The DREZ lesion as an effective treatment for chronic hypothetically post herpetic neuropathic pain. Case report and review of literature]. AB - The authors present a case of a 54-year-old woman with a 3-year history of chronic pain syndrome, probably of postherapeutic origin, with diffuse segmentary dermatome characteristics, both somatic and autonomic. The former were exemplified by a constant "burning" skin pain in the representation of Th8-LI dermatomes unilaterally, while the latter by a unilateral visceral pain within the abdominal cavity. Electrophysiological examination indicated a neuropathic origin of the pain, despite the lack of clinically evident sensory deficits and/or hypersensitivity. The pain was so intense that normal walking was difficult for the patient and ineffectiveness of her treatment made her suicidal. Since both pharmacological treatment (non-steroid analgesics, opioids, antidepressants, and anticonvulsants including gabapentin) and minimally invasive methods of treatment (blockades, thermolesions) failed to control pain, she was subjected to surgery. A right-sided DREZ lesion within the Th8-LI dermatomes resulted in a complete pain relief, both within the somatic and autonomic innervation projections, and in the patient's functional recovery. PMID- 14746253 TI - [Acute cervical epidural hematoma as a complication of anterior cervical C5-C6 diskectomy. A case report]. AB - A 32 year-old woman with cervico-brachial pain and progressive neurological disorders was admitted to the Department and Clinic of Neurosurgery and Neurotraumatology of the K. Marcinkowski University of Medical Sciences in Poznan. Neurological examination revealed right hemiparesis with muscular atrophy in the right upper limb, progressing in the past 3 months. MRI demonstrated relatively large herniations of C4-C5 and C5-C6 discs with spinal and nerve compressions. A surgery was carried out. Discectomy of C4-C5 and C5-C6 discs was performed using the anterior vertebral approach. After the removal of C4-C5 and C5-C6 discs and C5 vertebral body a fragment of the iliac bone strut was wedged between C4 and C6 vertebral bodies. The bone graft was fixed to C4 and C6 vertebral bodies with a titanium plate. Within 6 hours after the surgery the patient developed a complete loss of motor functions in the upper and lower limbs. MRI scan of the cervical spine showed a huge anterior epidural hematoma compressing the spinal cord. An emergency re-operation and removal of the hematoma was performed by the anterior approach. A complete neurological recovery followed during the first 24 postoperative hours. PMID- 14746254 TI - "The big C": it can't happen to me! PMID- 14746255 TI - President's address. "Researching for the Height of Excellence". PMID- 14746256 TI - Curing halitosis: the sweet smell of success. AB - Halitosis is socially incapacitating, physically damaging, and difficult to discuss. As the dentist is responsible for the oral cavity, it behooves the dental health provider to diagnose and treat halitosis if we are to offer total oral health care to our patients. While oral malodor has been recognized in the literature throughout our history, interest has recently increased in treating the problem and research is now providing the answers to this condition. This article summarizes some of the current research in managing halitosis. PMID- 14746257 TI - Contemporary permanent tooth dimensions, with comparisons to G.V. Black's data. AB - The tooth dimensions most commonly cited in the dental literature are those published by the famous restorative dentist G. V. Black late in the 19th century. We report here on the crown dimensions of a large series of contemporary North American Whites and show that Black's values differ significantly from modern values, but not in any consistent or predictable fashion. Black supplied no information about the nature of his sample. We suggest that it may be time to replace Black's data with modern tooth size standards. PMID- 14746258 TI - Preparing for the 2004 Georgia General Assembly. PMID- 14746259 TI - MAG's health policy office coordinates its efforts with public health and other community agencies. PMID- 14746260 TI - Moving backward. PMID- 14746261 TI - Current legal status of malpractice reform. PMID- 14746262 TI - Ten ways to attract new patients to your practice. PMID- 14746263 TI - The MDA health plan: we're all in it for the long haul. PMID- 14746264 TI - When does ethical advertising cross the line? PMID- 14746265 TI - Waste management regulations and dental amalgam: Part II. PMID- 14746266 TI - Oral pathology quiz: an unusual pericoronal cyst. The diagnosis was glandular odontogenic cyst, although focal areas consistent with dentigerous cyst were seen. PMID- 14746269 TI - Humicolous buthoid scorpions: a new genus and species from French Guiana. AB - A new genus and species of humicolous buthid scorpion are described on the basis of a single specimen collected in French Guyana. New considerations on the taxonomy and morphology of some micro-buthoid humicolous scorpions are proposed, mainly based on the study by scanning electron microscopy of the peg-shaped sensillae of the pectines. PMID- 14746268 TI - Injection of exogenous RNA in amphibian oocytes leads to RNA level fluctuations which are sensitive to cordycepin, an RNA chain elongation terminator. AB - Using an in vivo heterologous system to study the stability of Xenopus laevis RNA injected into axolotl (Ambystoma mexicanum) fertilized eggs, we have previously observed unexpected fluctuations in RNA level during early development [Andeol et al., Differentiation 63 (1998) 69-79]. In this study, we further characterize this phenomenon and establish its existence during axolotl and Xenopus oogenesis, suggesting a phylogenetically conserved mechanism. The phenomenon can occur with a variety of exogenous sense and antisense substrates. RNase protection experiments establish that most of the molecules have the same polarity as the initially injected RNA. In addition, trace amount of complementary RNA (cRNA) can be detected the injected samples. Cordycepin prevent increases in RNA levels indicating the involvement of an RNA synthesis. These results indicate the existence of an in vivo post-transcriptional RNA amplification mechanism during the early development of amphibians. PMID- 14746270 TI - [Structural variations of the eye linked to daily vertical migrations of Daphnia longispina]. AB - The role of the eyes, and more precisely that of the ommatidian pigments, in the control of daily rhythms of movement of Daphnia longispina is investigated. In the laboratory, under permanent light (LL), the pigmentary modifications observed are globally similar to those observed in situ, except in their timing, around dusk, LL cycles do not coincide with the ascent of daphnids but precede it. This is the expression of an endogenous free-running rhythm. Therefore, in D. longispina, an internal oscillator controls DVMs, which are circadian and not nycthemeral, and, in situ, illumination at dawn suffices to synchronize migrations on dark/light alternation. PMID- 14746271 TI - [Bioprotection mechanisms of the lentil plant by Rhizobium leguminosarum against Fusarium oxysporum f. sp. lentis]. AB - Living and heat-killed bacterial cells of Rhizobium leguminosarum protected totally lentil plants against infection by the pathogen Fusarium oxysporum MR 84. Culture filtrate of this rhizobacterium was also able to protect the plants to a high degree. However, when they were inoculated separately of the pathogen, living bacterial cells did not protect the plants whereas culture filtrate and killed bacterial cells protected them. These results suggest that Rhizobium cannot protect lentil plants without interaction with the pathogen, but the culture filtrate and the killed bacterial cells can protect them even in the absence of this interaction. It seems that the culture filtrate and the killed bacterial cells contain signals able to induce plant resistance. Those signals would be suppressed once Rhizobium is in contact with the plant. PMID- 14746272 TI - Replication of hexitol oligonucleotides as a prelude to the propagation of a third type of nucleic acid in vivo. AB - No backbone motif other than phospho-ribose and phospho-deoxyribose has been found in natural nucleic acids, currently restricting the molecular types of replicable biopolymers to DNA and RNA. With the aim of propagating and expressing a third type of nucleic acid in vivo, we assessed the replicability of polynucleotides with a phospho-hexitol backbone (HNA) in vivo and in vitro. Faithful polymerisation of up to four deoxynucleotides templated by hexitol oligonucleotides was established in vitro using DNA polymerase from Escherichia coli (PolA Klenow exo-fragment) and Thermus aquaticus (Taq polymerase). Condensation of up to three successive hTTPs (hexitol thymidine triphosphate) in responses to a pentameric hexitol template (hA)5 could also be demonstrated in vitro. Such a marginal HNA-dependent HNA polymerase activity of natural polymerases may be evolved in the future to catalyse in vitro amplification of HNA. The transmission of a two-codon-long genetic message carried on a hexameric hexitol template was also established using a selection screen for restoring thymidylate synthase activity in E. coli. These results exemplify the potential that can be explored by converting artificial substrates with natural enzymes in the field of informational polymer synthesis. PMID- 14746273 TI - Vocal similarities and social bonds in Campbell's monkey (Cercopithecus campbelli). AB - This study examined variations of social call structures in female Campbell's monkeys. The comparison of frequency modulations using a similarity index revealed that each individual presented one to three variants. Some variants were shared by two to several individuals, often associated with play. On the contrary, vocal divergence was observed in a socially isolated animal and a negative correlation appeared between similarity index and the frequency of avoidance. This preliminary study gives the first evidence of variant sharing between group members in primates. PMID- 14746274 TI - Bloodborne pathogen exposures and prevention in the perioperative environment. AB - The fear of exposure to bloodborne pathogens and disease transmission from patients to health care professionals or health care professional to patients is real. Perioperative nurses and other surgical personnel should be aware of the dangers and risks of exposure to these viruses. Patients may be infected with one of the currently recognized bloodborne viruses including Hepatitis B (HBV), Hepatitis C (HCV) and the Human Immunodeficiency Virus (HIV). It is imperative that accidental exposure during surgical procedures be avoided. The most common means of exposure are percutaneous and mucous membrane routes. The persistence of HBV, HCV and HIV infections, long incubation periods, and the likelihood of frequent asymptomatic carrier states remain continuing threats to the surgical team and makes it difficult to rely on a diagnosis. Infection prevention and control strategies to reduce the risk of exposures and prevent transmission are based on a number of approaches including engineering, administrative and works practice controls and the proper selection and use of personal protective equipment. PMID- 14746275 TI - Operation Smile China. PMID- 14746276 TI - [Adolescence: which solutions to better help our young people?]. PMID- 14746277 TI - [Nurses: a profession on all fronts. Interview by Paul Dauriac]. PMID- 14746278 TI - [The cancer plan of Jean-Francois Mattei]. PMID- 14746279 TI - [Classification of analgesics]. PMID- 14746280 TI - [Rotation of opioids]. PMID- 14746281 TI - [Acute (or chronic) intense pain. Oral administration impossible. Titration]. PMID- 14746282 TI - [Hepatitis C, there is much more to do...]. PMID- 14746284 TI - [Pheochromocytoma: a hypertensive tumor]. PMID- 14746285 TI - [Emergency management of cardiorespiratory arrest]. PMID- 14746286 TI - [Reflections on the education of patients with stomas: from myth to reality]. PMID- 14746287 TI - ["Handicapped persons: focus our attention on their potentials rather than their deficiencies. Interview by Marie-France Guerel]. PMID- 14746290 TI - Recent developments in privacy legislation. AB - In this article, Ruth Carey takes a critical look at recent privacy-of-personal information legislation drafted in three Canadian provinces--Ontario, Alberta, and British Columbia. The article begins with a historical overview of international legal instruments and other privacy guidelines, and the Canadian experience with privacy protection. It then critically analyzes the provincial initiatives in the context of the federal Personal Information Protection and and Electronic Documents Act and accepted privacy principles. The article goes on to highlight certain types of legislative provisions of particular interest to people with HIV/AIDS and those who advocate on their behalf. It concludes that the numerous legislative initiatives underway in Canada provide an opportunity to alter the public discourse around the virus, thereby improving the lives of people with HIV/AIDS. PMID- 14746291 TI - Supervised injection sites: Health Canada approves Vancouver pilot sites, but concerns remain. AB - The paperwork has been processed, and Health Canada says that two pilot supervised injection sites (SISs) in Vancouver can now proceed, but operational funding for the site to be located in the Downtown Eastside has yet to be secured. Meanwhile, tired of waiting for the sanctioned SISs to open, community groups in Vancouver have begun to operate unsanctioned sites. There are also concerns that even if operational funding for the Downtown Eastside site is obtained, the pilot SIS project is too small in scope to make a major impact on public health in the area. PMID- 14746292 TI - Health Canada unveils plan to distribute marijuana for medical use. AB - Under pressure from the courts, Health Canada reluctantly comes up with a distribution plan to provide dried cannabis and seeds to patients using medical marijuana. The plan has been greeted with considerable criticism PMID- 14746293 TI - Clinical trial on medicinal use of marijuana cancelled. AB - Despite concerns expressed about the lack of scientific data on the safety and efficacy of marijuana, no clinical trials on the medical uses of marijuana have been completed in Canada. PMID- 14746294 TI - Ottawa moves to decriminalize small amounts of marijuana. AB - Making good on a promise made in December 2002, the federal government has tabled a bill that would decriminalize possession of small amounts of marijuana. However, the bill would also toughen penalties for growing large amounts. The bill is part of a renewed Canada Drug Strategy that emphasizes the need to discourage cannabis use. PMID- 14746295 TI - Commons committee recommends significant increase in funding for AIDS strategy. AB - A House of Commons committee says that current funding for the Canadian Strategy on HIV/AIDS should be more than doubled. It calls for designated funding for inmates and First Nations and Inuit peoples, and for HIV vaccine research. Finally, it says that more federal government departments need to become involved in the response to AIDS. PMID- 14746296 TI - Ontario: pressure from stakeholders leads to improved access to selected drugs. AB - A campaign spearheaded by the Canadian Treatment Action Council, and involving other disease groups, has convinced the Ontario government to cover the costs of vaccines for HIV-positive children and to speed up access to other subsidized drugs. PMID- 14746297 TI - Class orders now possible under Ontario's public health legislation. AB - In April 2003, the Ontario Legislature amended the province's public health legislation as part of a package of amendments related to the recent outbreak of Severe Acute Respiratory Syndrome (SARS). Although the amendments to the Health Protection and Promotion Act (HPPA) were clearly designed to address emergency situations like SARS, they may have unintended and negative consequences for people living with HIV/AIDS. PMID- 14746298 TI - Commons committee calls for more leadership and resources to fight AIDS in Africa. AB - The House of Commons Standing Committee on Foreign Affairs and International Trade issues a report that concludes that the magnitude and urgency of the AIDS crisis in sub-Saharan Africa requires additional action and resources from the federal government. PMID- 14746299 TI - Saskatchewan agrees to cover new HCV and HIV drugs. PMID- 14746300 TI - Still no HIV/AIDS strategy in New Brunswick. PMID- 14746301 TI - Health Canada announces public consultations on proposed health legislation. PMID- 14746302 TI - Woman sues physicians over failure to offer prenatal HIV test. PMID- 14746303 TI - US international AIDS funding will also advance a moral agenda. PMID- 14746304 TI - Zambia: judges workshop addresses HIV/AIDS. PMID- 14746305 TI - UK report on migration and HIV rejects mandatory testing. PMID- 14746306 TI - Caribbean nations hold first training workshop on AIDS laws and policies. PMID- 14746307 TI - Canada: national summit on "Global health is a human right"! AB - Held in Ottawa on 21-22 May 2003, a national summit on "Global Health is a Human Right!" brought together 86 people representing over 60 Canadian civil society groups interested in joint advocacy on realizing the human right to health in developing countries. The summit focused particularly on addressing the global crises of communicable diseases such as HIV/AIDS, tuberculosis, and malaria. PMID- 14746308 TI - [Development and problem of soft tissues neoplasms]. PMID- 14746309 TI - [Approaches in molecular pathology of some gynecological tumors]. PMID- 14746311 TI - [Rethinking about the prevalence of SARS, recent work on SARS prevention and control]. PMID- 14746310 TI - [Application of proteomics in digestive system tumours]. PMID- 14746312 TI - [Present status and future of basic research on breast cancer]. PMID- 14746313 TI - Cerebral vasoconstriction and stroke after use of serotonergic drugs. PMID- 14746314 TI - Polio-endemic countries pledge to boost eradication efforts in 2004. But whether the disease can be eradicated will depend on situation in Nigeria. PMID- 14746315 TI - Canada's new prime minister vows to cut surgery wait lists. Martin considers point-scoring systems to rationalise and speed scheduling. PMID- 14746316 TI - WHO praises Bam response but warns of disease. PMID- 14746317 TI - FDA demands more information about breast implants. PMID- 14746318 TI - Many countries not on target to reach millennium goals. PMID- 14746319 TI - Japan steps up SARS plans. PMID- 14746320 TI - EU deplores US execution of mentally ill prisoner. PMID- 14746321 TI - Violence prevention receives international attention. PMID- 14746322 TI - Education for HIV orphans. PMID- 14746323 TI - Better breastfeeding. PMID- 14746324 TI - Food for AIDS. PMID- 14746325 TI - Ugandan officials negotiate Global Fund grants. Government limits on health sector spending may jeapordise funding agreement. PMID- 14746326 TI - Deprivation, disease, and the ethics of international research. PMID- 14746327 TI - Health authority cervical screening recall policies and time since last smear: a retrospective cohort analysis in the north west England. AB - OBJECTIVES: To evaluate the impact of health authority cervical screening programmes upon the the actual screening interval for women attending as a result of routine recall and to assess the impact of general practice recall policies on attendance for smears. SETTING: Nine health authorities constituting the Lancashire and Greater Manchester zones of the North West region on the National Health Service. METHODS: Information was obtained on the invitation and recall policies of each health authority. A retrospective analysis of information held on cervical screening databases ("Exeter" computer systems was performed. The proportions of smears, classified by a screening interval of "unscheduled", "on time" or "overdue", in the different health authority recall programmes were compared. General practices in one health authority (operating five-yearly recall) were contacted to determine their current practice screening policy. RESULTS: Women who were invited as part of a three-yearly recall programme were more likely to attend for smears at intervals of 3-5 years than those on a five yearly recall programme (68% vs 47%). Those health authorities operating three yearly recall that issued recall invitations to women more than four weeks before the date the smear was due had higher rates of "unscheduled" smears. In the one health authority (operating a five-yearly recall programme) in which we determined general practice screening policy, 62% of the general practitioners (GPs) were operating their own three-yearly recall policy; these GPs had a higher proportion of the women who attended for smears "on-time" (51%) than did those who relied on the health authority five-yearly recall procedure (44%). CONCLUSIONS: Despite the cervical screening programme being a national programme, there is scope at a local level for considerable variation in the interpretation of national guidance. This variation effects the extent to which programmes comply with the requirement to screen all women at an interval of 3-5 years. PMID- 14746328 TI - [Development and property study of zirconia toughened nano-composite alumina ceramic powder for dental application]. AB - OBJECTIVE: To prepare zirconia toughened nano-composite alumina ceramic powder for dental application. Physical and chemical property of the prepared material were tested, and the effect of development technology on composite powder was also studied in this study. METHODS: Nano-composite alumina powder was prepared by surface-induced precipitation method. The effect of pH value and dispersing agent content on volume of alumina suspension sediment was recorded. The effect of ultrasonic time on agglomeration was measured also. X ray diffraction (XRD) was used to analyze powder phase before and after the stabilizer was added. Scanning electronic microscope (SEM) was applied for characterizing the specimen. RESULTS: The dispersion was better at pH=9 and wt (dispersing agent) = 0.2% approximately 0.3%. Selecting proper ultrasonic time can decrease the agglomeration of powders and lower the average particle size. XRD analysis indicated that the phase composition of the prepared nano-composite ceramic powder was shown as alpha-Al2O3, t-ZrO2 and a small amount of m-ZrO2 after the addition of stabilizer. Through SEM observation, nanometer-sized ZrO2 particles (80 approximately 100 nm) were uniformly located on the surface of submicrometer alumina grains. CONCLUSIONS: By choosing appropriate preparation method, weakly agglomerated powders with fine particle size can be obtained. The zirconia part of nano-composite powder was transmitted to partially stabled zirconia after the use of stabilizer. PMID- 14746329 TI - [The research of using TiN nanometer film to improve the anticorrosive property of FeCrMo alloy]. AB - OBJECTIVE: The aim of the study is to improve the anticorrosive property of the dental FeCrMo soft magnetic alloy covered with TiN film obtained by ion beam assisted deposition (IBAD) technology in oral environment. METHODS: The magnetic force of the FECrMo soft magnetic alloy after TiN film treated were measured by Instron test machine. An advanced electro-chemical method was used to measure the electric potential of corrosion (Ecorr), passive potential (Ep), passive current density (Ip), current density of corrosion (Icorr), polarization resistance (Rp), of FeCrMo soft magnetic alloy in simulated oral environment before and after surface modification. RESULTS: There were no statistic changes of the magnetic force in 4 groups after alloy with TiN film treated. Comparing with the alloy without surface modified, the Ecorr, Rp of FeCrMo soft magnetic alloy was obviously higher, and the Icorr, Ip and Ep were obviously lower. CONCLUSIONS: The anticorrosive property of the dental FeCrMo soft magnetic alloy with TiN film is better than that without modified. PMID- 14746330 TI - [The study of hBMP-4 gene modified tissue-engineered bone]. AB - OBJECTIVE: Bone marrow stromal cells (bMSCs) of rabbits transferred with mammalian hBMP-4 expression plasmid were used to construct tissue-engineered bone. Gene therapy combined with tissue-engineering technique was explored to further improve osteogenesis. METHODS: pEGFP-hBMP-4 plasmid was constructed by subcloning technique. bMSCs were then transferred with either pEGFP-hBMP-4, pEGFP plasmid by lipofectamine or left uninfected in vitro. The cells from the 3 groups were combined with natural non-organic bone (NNB) to construct tissue-engineered bones, which were subcutaneously implanted into nude mice (6 implants per group) for 4 weeks. Specimens were evaluated through histological and computerized new bone formation analysis. RESULTS: pEGFP-hBMP-4 plasmid was successfully constructed. bMSCs could attach and proliferate on the surface on NNB. In vivo experiment showed that new bone formation in pEGFP-hBMP-4 group was higher than those of the control groups. CONCLUSIONS: Tissue-engineered bone using hBMP-4 gene modified bMSCs might be an ideal alternative for the repair of bone. PMID- 14746331 TI - [A study of injectable autogenous tissue-engineered bone]. AB - OBJECTIVE: To investigate the utilization of carrier for delivering osteoblasts and creating autogenous bone tissue in ectopic site of animal via injection. METHODS: Bone marrow cells harvested from iliac bone of New Zealand rabbits were induced to differentiate into marrow stromal osteoblasts. The osteoblasts were mixed with 1.5% alginate sodium solution to generate osteoblasts/alginate composites with final cellular density of 4 x 10(9)/L. Calcium chloride was used as cross-linking agent to gel aqueous alginate solution. The marrow stromal osteoblasts/alginate composites were injected into the dorsal subcutaneous tissue of rabbits with autogenous cells transplantation. The samples were examined with X-ray and histological analysis. RESULTS: Four, eight and twelve weeks after injection, the hard knobbles were easily palpated under the dorsal skin of animals. On X-ray photograph the samples showed calcified image with more density than surrounding soft tissue, new bone formation was observed in the osteoblasts/alginate composites in histological analysis. The osteogenesis was in association with regenerated hematopoietic bone marrow. CONCLUSIONS: These results demonstrate that new bone tissue could be created through the injection of alginate sodium treated with autogenous marrow stromal osteoblasts. PMID- 14746333 TI - Proceedings of the Second International Conference on Male Mediated Developmental Toxicity. Montreal, Canada. June 2001. PMID- 14746332 TI - New trends in cancer for the 21st Century. Proceedings of the International Symposium on Cancer. Valencia, Spain. November 10-13, 2002. PMID- 14746334 TI - Human gene therapy: current opportunities and future trends. Proceedings of the Ernst Schering Foundation Workshop. Berkeley, California, USA. October 2-4, 2002. PMID- 14746335 TI - Novel approaches for the discovery of anticancer agents. Abstracts from a symposium. Freiburg, Germany. June 18-21, 2003. PMID- 14746336 TI - Proceeds and abstracts of the Middle Eastern Mediterranean Summer Institute on Drug Use. Pavia, Italy. May 23-26, 2001. PMID- 14746337 TI - Proceedings of the International Conference for the Bone and Joint Decade 2000 2010. Tokyo, Japan. April 17-19, 2002. PMID- 14746338 TI - The educator's guide to diabetes resources. PMID- 14746339 TI - [Advances in the study of Atractylodes macrocephala Koidz]. PMID- 14746340 TI - First and second trimester antenatal screening for Down's syndrome: the results of the Serum, Urine and Ultrasound Screening Study (SURUSS). PMID- 14746341 TI - Possible mechanism on enhanced carcinogenesis of genotoxic carcinogens and unsolved mechanisms on lesser carcinogenic susceptibility to some carcinogens in rasH2 mice. AB - The rasH2 mice are hemizygous transgenic mice carrying the human prototype c-Ha ras gene with its own promoter region, and have been used in 6-month short-term carcinogenicity tests for pharmaceutical drugs in accordance with the recommendation of the International Conference on Harmonization of Technical Requirements of Pharmaceuticals for Human Use (ICH). Based on the validation studies, it has been recognized that they are very susceptible to genotoxic carcinogens. To elucidate the mechanism of the enhanced carcinogenesis, spontaneous and chemically induced tumors in rasH2 mice have been subjected to molecular analyses, but the results have thus far been equivocal. This article focuses on the possible molecular mechanism of enhanced carcinogenesis in rasH2 mice, based on the results of a search in the literature. In addition, there are several reports suggesting lesser carcinogenic susceptibility of rasH2 mice to some carcinogens: Malignant lymphomas were induced by treatment with phenolphthalein in heterozygous p53 knockout mice, but not in rasH2 mice, and ethinylestradiol, uterine tumor promoter, resulted in depression of uterine proliferative lesions in rasH2 mice. In this review, the possible mechanisms of why rasH2 mice were less sensitive for these carcinogens are also discussed. PMID- 14746342 TI - Effects of maternal exposure to diethylstilbestrol on the development of the reproductive system and thyroid function in male and female rat offspring. AB - Diethylstilbestrol (DES) was administered subcutaneously at 1.5 or 15 microg/kg/day (DES 1.5 group, DES 15 group) to pregnant SD rats daily on days 7 21 of gestation to investigate its effects on the development and functions of the reproductive system and thyroid gland in their offspring. Of the 11 pregnant rats in the DES 15 group, only one delivered a live pup. Rat pups in the DES 1.5 group were autopsied at 1, 3, or 6 weeks after birth. In the DES 1.5 group, the plasma T4 concentrations at all weeks of age at autopsy were significantly increased, the TSH concentration at 6 weeks of age was also significantly increased, and the height of thyroid follicular epithelial cells was increased at 3 weeks. The testosterone concentration in the DES 1.5 group at 6 weeks was significantly decreased and the plasma LH concentration was increased. The DES treatment increased the plasma FSH concentration in female pups at 3 weeks, increased the percentages of primary and secondary ovarian follicles, and decreased the percentage of primordial follicles, but did not influence the timing of the vaginal opening or the onset of the estrous cycle. These observations indicate that prenatally administered DES increases thyroid function, and has an inhibitory effect on testicular function and a promoting effect on female reproductive function. PMID- 14746343 TI - Formation of highly analgesic morphine-6-glucuronide following physiologic concentration of morphine in human brain. AB - 3H-Morphine at physiologic concentration was metabolized in vitro to its 3- and 6 glucuronides by human brain homogenate. Recombinant UGT2B7, one of the UDP glucuronosyltransferase (UGT) isoforms, is able to glucuronidate the 3- and 6 hydroxy groups of morphine at nanomolar concentrations. These results suggest that endogenous morphine is converted to its 6-glucuronide, a more highly analgesic substance than the parent compound, to suppress effectively pain symptoms in humans. PMID- 14746344 TI - A comparative study on the reliablility of toxicokinetic parameters for predicting hepatotoxicity of DDT in rats receiving a single or repeated administration. AB - A comparative study on the reliability of toxicokinetic parameters for predicting hepatotoxicity was conducted in male F344 rats receiving a single (106 mg/kg by gavage) or 7-day repeated (1000 ppm in feed, 97 mg/kg/day) administration of p,p' DDT. DDT was selected as the test substance because it is known as a hepatotoxic agent and its metabolic pathway is well documented. Concentrations of DDT and its metabolites (DDE and DDD) in the plasma, brain, and liver were measured at various time intervals during the study and the results were compared with the area under the concentration-time curve (AUC) in relation to hepatotoxic response. Increases in the absolute and relative (ratio to body weight) liver weights were observed as a typical toxic response after a single or repeated exposure to DDT. The coefficient (R2) of correlation between the increases in the relative liver weight and the concentrations or AUC of DDT and its metabolites in the plasma and liver was estimated. The values of R2 between the relative liver weight and AUC of DDT or the total DDT (T-DDT) in the plasma and liver were found to be more consistent and higher than those with their concentrations in the repeated dose study. In addition, the R2 values in correlation with their AUCs after a single exposure were lower than those in the repeated dose study. These results indicate that the AUC of DDT or T-DDT in the plasma and liver would be more reliable than their concentrations for predicting hepatotoxicity caused by DDT, especially in the repeated dose study. PMID- 14746345 TI - Comparison of poultice-type and tape-type patches containing ketoprofen on human skin irritation. AB - Although the patch test for visual skin observation is widely used clinically, it does not allow the mechanisms of side effects to be assessed. In this study, we examined poultice-type KP801 and tape-type KP-T patches containing ketoprofen. The parameters to measure side effects on skin were peeling intensity, amount of stripped stratum corneum, skin moisture and redness of skin color under various mechanical conditions. Since the amount of stripped stratum corneum with the tape type KP-T patch was higher than with the poultice-type KP801 patch, the bio adhesive strength of the latter was concluded to be lower. A clear relationship exists between the amount of stripped stratum corneum and skin moisture after tape-type patch removal, but this was not found with the poultice-type patch because of its hydration effects. Peeling intensity, one parameter to predict pain at the time of patch removal, was higher with the KP-T. As for mechanical conditions, when the patch is removed, it is important to remove it as slowly as possible and horizontally, and to avoid any rise in skin temperature. Finally, when a patch is applied to a region with little skin moisture, the amount of stripped stratum corneum may increase accordingly. PMID- 14746346 TI - Differential modulation of muscarinic receptors in the rat brain by repeated exposure to methyl parathion. AB - The neurochemical and behavioral effects of repeated subdermal administration of methyl parathion (MP) at low doses were investigated. Adult male rats were treated repeatedly with either vehicle or MP subcutaneously (3 mg/kg/day) and observed for the signs of toxicity during the treatment period. The toxic sign, tremor, reached maximum right after 9th injection in MP-treated rats, and declined thereafter. Animals were sacrificed and brains were taken 1 week or 3 weeks after the daily treatment for measurement of acetylcholinesterase (AChE) activity and binding of radioligands, [3H]QNB (nonselective), [3H]pirenzepine (M1 selective), and [3H]AF-DX384 (M2-selective) to muscarinic receptors. With this treatment regimen, the AChE activity in the blood dropped quickly and maintained at 30% of the control level after 6 injections. After 3 weeks of treatment, MP caused 80-90% AChE inhibition and substantial reductions in [3H]QNB binding (9 33%), [3H]pirenzepine binding (9-22%) and [3H]AF-DX384 binding (6-38%) in different brain regions, including striatum, hippocampus, frontal cortex, thalamus and midbrain. After 1 week of treatment, the inhibition of AChE in brain regions was from 54 to 74%, whereas receptor densities were only marginally affected in a few regions. The timing of the changes in receptor population correlates well with the changes in behaviors during the repeated MP exposure. Our findings suggest that down-regulation of muscarinic receptors plays a role in the development of tolerance to MP. And, the regulations of muscarinic receptors were different among receptor subtypes and brain regions. PMID- 14746347 TI - Mechanism for shortening PT and APTT in dogs and rats--effect of fibrinogen on PT and APTT--. AB - The possible mechanisms for shortening prothrombin time (PT) and activated partial thromboplastin time (APTT) were investigated using citrated plasma from rats and dogs in vitro, especially focusing on increased fibrinogen concentrations. When purified canine fibrinogen was added to citrated canine plasma at final concentrations of 2, 4 and 8 mg/mL, PT and APTT were significantly shortened. The increased concentrations of clottable fibrinogen in the test system were confirmed by markedly shortened thrombin time (TT). In citrated rat plasma, while purified rat fibrinogen had no effect on PT or APTT at final concentrations of 2, 4 and 8 mg/mL, it did shorten TT. These results suggest that an increased concentration of fibrinogen is a possible mechanism to shorten PT and APTT in dogs, but not in rats. PMID- 14746348 TI - Four-week oral toxicity study of 1-carboxy-5,7-dibromo-6-hydroxy-2,3,4 trichloroxanthone (HXCA), an impurity of Phloxine B, in F344 rats. AB - This study was designed to evaluate and characterize any subacute toxicity of 1 carboxy-5,7-dibromo-6-hydroxy-2,3,4-trichloroxanthone (HXCA), an impurity of Phloxine B (Food Red No. 104 in Japan, D&C Red No. 28 in the USA), when administered to both sexes of F344 rats at dietary levels of 0 (control), 0.005, 0.05 and 0.5%. During the study, the treatment had no effects on clinical signs, survival, urinalysis or ophthalmology. Hematology, blood biochemistry, gross pathology, organ weights, organ to body weight ratios and histopathology exhibited no differences of toxicological significance between control and treated rats. Reactions to treatment may be summarized as follows: there was a tendency for increased food and water consumption and decreased food efficiency in both sexes of the 0.5% group. Thus, these results indicated the no-observed adverse-effect level (NOAEL) of HXCA to be 0.05% (39.3 mg/kg/day for males, and 41.0 mg/kg/day for females). PMID- 14746349 TI - A novel model of continuous depletion of glutathione in mice treated with L buthionine (S,R)-sulfoximine. AB - L-buthionine (S,R)-sulfoximine (BSO), an inhibitor of glutathione (GSH) synthesis, was administered to mice via drinking water for 14 days in order to establish an animal model with continuously depleted levels of GSH. No toxicity was observed at 20 mM BSO, even though a significant decrease in liver weight was observed at 30 mM BSO. GSH levels in the liver, kidney, brain, lung, heart, spleen, pancreas, small intestine, large intestine, skeletal muscle, plasma and blood cells from mice given 20 mM of BSO were all less than those from the control mice continuously throughout a 24-hr period. The ratios of the GSH levels to that of the control were 46.4% and 16.7% in the liver and kidney, respectively, suggesting a decrease in GSH conjugation activity in vivo by GSH depletion. Liver cytochrome P450 content and UDP-glucuronosyltransferase activity to p-nitrophenol were not influenced by the BSO dosing. To confirm the adequacy of this GSH-depletion model, 0.125 or 0.25% of acetaminophen (APAP) was administered via diet to this model for 14 days. Nine out of the ten mice given both 20 mM BSO and 0.25% APAP died on Day 2, and remarkable necrosis was observed in the hepatocytes and renal tubular epithelium. Moreover, focal necrosis of hepatocytes with proliferation of fibroblasts was observed on Day 15 in some mice coadministered 20 mM BSO and 0.125% APAP. However, no toxicity was observed in mice given APAP alone. Based on these results, a mouse given 20 mM of BSO via drinking water for 14 days was concluded to be an animal model with continuously depleted levels of GSH in various organs without toxicity. This model shows high susceptibility to toxicity induced by chemicals which are metabolized to electrophilic and reactive metabolite(s), such as APAP. PMID- 14746350 TI - Thirteen-week repeated dose toxicity study of wormwood (Artemisia absinthium) extract in rats. AB - Wormwood, Artemisia absinthium, is a very bitter plant, and its extract has been used as food additives such as seasonings for food and drinks. A 13-week repeated dose toxicity study of wormwood extract was performed in both sexes of Wistar Hannover (GALAS) rats. Rats were divided into 4 groups consisting of 10 males and 10 females each, and were given water containing 0, 0.125, 0.5, or 2% wormwood extract. All rats had survived at the end of the study, and no changes indicating obvious toxicities that are attributable to the treatment of wormwood extract were observed in the body weights, hematological and serum biochemical examinations, organ weights, and histopathological examinations. Based on the results of the present study, the NOAEL (no-observed-adverse-effect-level) of wormwood extract of Wistar Hannover rats was estimated to be 2% (equivalent to 1.27 g/kg/day in males and 2.06 g/kg/day in females) or more. PMID- 14746351 TI - A classification of Black-American self-esteem. PMID- 14746352 TI - Patient attitudes regarding healthcare utilization and referral: a descriptive comparison in African- and Caucasian Americans with chronic pain. AB - The purpose of this study was to evaluate healthcare utilization and referral patterns for pain management services in a racially and ethnically diverse population. A study-specific mail survey was directed at African- (N=324) and Caucasian Americans (N=300) receiving chronic pain treatment at a tertiary care pain center to address their healthcare access, referral, and utilization patterns. Overall, 46% (N=286) responded, with the majority of respondents being Caucasian Americans (57%) and women (68%). The majority (58%) reported asking their physicians to refer them to a pain physician. African Americans were more likely to report that chronic pain was a major reason for financial problems. They made significantly more visits to the emergency room for pain care. African Americans agreed more that ethnicity and culture affected access to healthcare and pain management. They also tended to agree more than Caucasian Americans that pain medication could not control pain. These results demonstrate significant differences in healthcare utilization, access, and attitudes amongst African- and Caucasian Americans receiving chronic pain management. In light of the socioeconomic and health consequences of chronic pain, these results suggest the need for further studies addressing variability in pain care access and utilization in diverse populations. PMID- 14746353 TI - Racial differences in hepatitis B and hepatitis C and associated risk behaviors in veterans with severe mental illness. AB - Racial differences in the seroprevalence of and risks for hepatitis B (HBV) and hepatitis C (HCV) were examined in military veterans with severe mental illnesses (SMI). Participants (376; 155 Caucasian, 221 African American) were inpatients at a Veterans Affairs (VA) psychiatric unit in Durham, N.C., from 1998 to 2000. Prevalence rates of HBV and HCV were 21.3% and 18.9%, respectively. African Americans had a higher HBV seroprevalence than did Caucasians: 27.6% versus 12.3%; odds ratio (OR) 2.73; 95% confidence interval (CI)=1.55, 4.79. Although not statistically significant, HCV seroprevalence was also higher for African Americans than it was for Caucasians: 21.3% versus 15.5%; OR=1.47; 95% CI=0.86, 2.53. No racial difference was observed for injection drug use (IDU), the strongest risk indicator for both HBV and HCV. Multivariable analyses indicated that African-American race, IDU, and multiple sex partners in the past six months were related to an increased risk of HBV, whereas IDU and smoking crack cocaine were both independently related to an increased risk of HCV. Thus, veterans with SMI--particularly African-American veterans--have high rates of HBV and HCV infection. African-American veterans have significantly higher rates of HBV than do Caucasian veterans, which persist after controlling for prominent risk behaviors. PMID- 14746354 TI - The relationship between motivation to volunteer, gender, cultural mistrust, and willingness to donate organs among Blacks. AB - The relationship between motivation to volunteer, gender, cultural mistrust, and the willingness of blacks to donate their organs, as well as the organs of relatives, was explored. Participants consisted of 107 black students attending a university located in the southwest. All participants were given the Volunteer Functions Inventory (VFI), Cultural Mistrust Inventory (CMI), Organ Donation Questionnaire (ODQ), and a background information questionnaire. It was found that individuals with low scores on the VFI and high scores on the CMI were less willing to consent to donating their organs. Also, females and individuals with high CMI scores were less willing to permit the recovery of organs from relatives. Some theoretical and applied implications for mental health professionals are suggested. PMID- 14746355 TI - Surgical treatment of obesity: a review. AB - Obesity is a chronic disease due to excess fat storage, a genetic predisposition, and strong environmental contributions. This problem is worldwide, and the incidence is increasing daily. There are medical, physical, social, economic, and psychological comorbid conditions associated with obesity. There is no cure for obesity except possibly prevention. Nonsurgical treatment has been inadequate in providing sustained weight loss. Currently, surgery offers the only viable treatment option with longterm weight loss and maintenance for the morbidly obese. Surgeries for weight loss are called bariatric surgeries. There is no one operation that is effective for all patients. Gastric bypass operations are the most common operations currently used. Because there are inherent complications from surgeries, bariatric surgeries should be performed in a multidisciplinary setting. The laparoscopic approach is being used by some surgeons in performing the various operations. The success rate--usually defined as >50% excess weight loss that is maintained for at least five years from bariatric surgery--ranges from 40% in the simple to >70% in the complex operations. The weight loss from surgical treatment results in significant improvements and, in some cases, complete resolution of comorbid conditions associated with obesity. Patients undergoing surgery for obesity need lifelong nutritional supplements and medical monitoring. PMID- 14746356 TI - Addressing multiple breast cancer risk factors in African-American women. AB - This pilot study explored the acceptability and feasibility of and estimated the effectiveness of a weight loss/breast health intervention designed to reduce breast cancer risk in African-American women ages 35-65. The study had a one group repeated-measures design and took place in a community setting. Forty-four African-American women were recruited, 35 completed the program, and 30 returned for the one-year follow-up. The pilot intervention was three weeks in duration and included twice-weekly exercise classes and weekly active learning seminars that addressed weight loss, breast health, healthy eating, and leading an active life. Measures included those of behavior related to diet, physical activity, and breast health. Satisfaction questionnaires and focus groups were also used to assess acceptability and cultural competency. Statistical analyses included Paired t-tests and Wilcoxon signed ranks tests. Significant results postintervention showed improved physical activity, dietary, and breast health behaviors. Results suggest the acceptability, feasibility, and effectiveness of this comprehensive weight/loss breast health program in reducing multiple breast cancer risk factors among African-American women. PMID- 14746357 TI - Survival of U.S. Black and White patients with squamous cell cancer of the esophagus. AB - Using data from 11 population-based cancer registries on 1,125 black and 2,392 white patients diagnosed with squamous cell carcinoma of the esophagus in 1992 1998, black-white differences in the relative survival rate (RSR)-which adjusts for mortality in the general population-were large only for localized-stage cancer. Within localized stage, black-white differences in RSR were smaller among patients without pathologic review of regional lymph nodes (RLNs). The low frequency of pathologic review of lymph nodes (8% of blacks and 12% of whites) among patients coded as "localized stage" indicates that staging was based on clinical tests (not recorded in cancer registries) and cancer-directed surgery (which was less frequent in blacks than whites). In Cox proportional hazards regression models, including cancer-directed surgery, along with demographic characteristics, the relative risk of death (hazard ratio) for all blacks versus whites was only 1.13 for all stages but 1.31 in a model with only localized-stage patients. Studies are needed on the extent of radiographic, endoscopic and other techniques used to assess stage in black versus white patients. The low survival rates for both blacks and whites emphasize the need for improved treatment and primary prevention efforts. PMID- 14746358 TI - Intestinal parasitoses. AB - BACKGROUND: Intestinal parasitoses is a clinical problem in the developing world and severe parasitaemia may be associated with retroviruses. OBJECTIVE: Studies on intestinal parasitoses were conducted in Dominica, and the health implications in an HTLV-1 endemic area were discussed. METHOD OF STUDY: A retrospective study of data of stool samples analysed at the parasitology unit of the medical laboratory services of Princess Margaret Hospital, Dominica, was conducted in January-December 1999. RESULTS: Parasites were found in 393 out of 3,752 stool samples (10.47%). The main parasites were Entamoeba coli, 1.4% (51/3,752); hookworm, 1.5% (56/3,752); Giardia lamblia, 1.4% (51/3,752); Strongyloides stercoralis, 1.0% (37/3,752); Ascaris lumbricoides, 0.8% (28/3,752); and Trichuris trichiura, 0.9% (34/3,752). CONCLUSION: Intestinal parasites are still endemic in Dominica, but significant reduction in prevalence has occurred over the last two decades. PMID- 14746359 TI - Race and sexual identity: perceptions about medical culture and healthcare among Black men who have sex with men. AB - Black men who have sex with men (BMSM) in the United States are disproportionately affected by HIV. Using a qualitative approach, the authors describe the healthcare experiences of BMSM in New York State and Atlanta, GA, exploring the social issues that influence barriers to care, communication, and adherence in medical settings. Racial and sexual discrimination socially displace BMSM, and are often compounded by negative encounters within medical institutions. The internalization of these experiences influences healthcare utilization, HIV testing, communication, and adherence behaviors among members of this population. Increasing the number of ethnic and sexual minority providers, expanding current definitions of cultural competency curricula at academic institutions, targeting future research efforts on BMSM, and improving the structural and communication barriers within healthcare settings should be incorporated into our HIV prevention and routine healthcare interventions for BMSM. PMID- 14746360 TI - Prevention and treatment of excessive dermal scarring. AB - Today, wound management to avoid excessive scar formation is increasingly important, especially in populations with Fitzpatrick 3 or higher skin pigmentation. Medical science and industrial development are devoting more effort toward understanding and offering better therapy to control scars. However, advances in scar management have been hampered by the confusing or ambiguous terminology. There is no consensus on what amount of post-traumatic skin scar formation is "normal" and what should be considered "hypertrophic". In the World Health Organization's ICD-9, there is no diagnostic code for hypertrophic scar- only keloid is listed. Yet, the medical and scientific literature distinguishes them as different conditions. Our experience suggests that the diagnosis of keloid disease is greatly over-rendered. For black patients, an elevated scar seems, by default, diagnosed as keloid by most. This confusion results in inappropriate management of scar formation, and occasionally contributes to decision making related to elective or cosmetic surgery. Given that patients are expecting better outcomes from wound care today than in the past, this review article attempts to capture the essential biological factors related to wound scar production and discusses treatment options and indications used by the authors. PMID- 14746361 TI - A case of intestinal-type sinonasal adenocarcinoma. AB - Intestinal type sinonasal adenocarcinomas are a rare malignancy of the nasal cavity and paranasal sinuses. Its association with lengthy occupational exposure to dust has been well documented, but sporadic cases have been reported. Diagnostic imaging is essential to exclude metastatic disease from the colon and may be supported by immunohistochemical studies of biopsy specimens. Treatment consists of surgery, radiation, or both, but its effect on prognosis is heavily influenced by the stage at which the disease presents. PMID- 14746362 TI - Right-sided chest-lead abnormalities on EKG in acute pulmonary embolism: ST elevation. PMID- 14746363 TI - Digital imaging. PMID- 14746364 TI - CD/DVD technology. AB - For storage of large amounts of data, there are many options to consider, including both CD and DVD methods. Given that these are now becoming very cheap alternatives, and that we work in a specialty which generates large volumes of digital data, it is now obvious why CD and DVD burning is of interest to radiologists. These technologies can most assuredly be of great benefit to both our professional and personal lives. PMID- 14746365 TI - Radiology teaching presentation tools. PMID- 14746366 TI - Beyond PACS: getting images to referring physicians. AB - The digital imaging hospital has the potential advantage that imaging studies are available anytime, and anywhere they are needed, with negligible loss. Fulfilling this promise in reality, however, presents challenges as described in this paper. No single best formula for successful deployment exists, since each institution is unique, and its medical staff may have varying preferences. This requires a collaborative team approach to design a solution acceptable to all parties, inside and outside of radiology. We have attempted to identify the significant issues. In addition, hardware and software technology continuously evolve and improve. What is clear is that all institutions will adopt this technology for the reasons described. The successful implementations will be those that are collaboratively and thoughtfully designed. User training of all members of the healthcare team involved with image study access is essential. PMID- 14746367 TI - Handhelds in radiology. PMID- 14746368 TI - Hypocretin administration as a treatment for human narcolepsy. PMID- 14746369 TI - Sleep deprivation and circulatory control. PMID- 14746370 TI - Does sleep deprivation worsen mild obstructive sleep apnea? PMID- 14746371 TI - Masticatory movements during sleep: some food for thought. PMID- 14746372 TI - Anandamide enhances extracellular levels of adenosine and induces sleep: an in vivo microdialysis study. AB - STUDY OBJECTIVES: The principal component of marijuana, delta-9 tetrahydrocannabinol increases sleep in humans. Endogenous cannabinoids, such as N-arachidonoylethanolamine (anandamide), also increase sleep. However, the mechanism by which these molecules promote sleep is not known but might involve a sleep-inducing molecule such as adenosine. Microdialysis samples were collected from the basal forebrain in order to detect levels of adenosine before and after injection of anandamide. DESIGN: Rats were implanted for sleep studies, and a cannula was placed in the basal forebrain to collect microdialysis samples. Samples were analyzed using high-performance liquid chromatography. SETTINGS: Basic neuroscience research laboratory. PARTICIPANTS AND INTERVENTIONS: Three month-old male F344 rats. At the start of the lights-on period, animals received systemic injections of dimethyl sulfoxide (vehicle), anandamide, SR141716A (cannabinoid receptor 1 [CB1] antagonist), or SR141716A and anandamide. One hour after injections, microdialysis samples were collected (5 microL) from the basal forebrain every hour over a 20-minute period for 5 hours. The samples were immediately analyzed via high-performance liquid chromatography for adenosine levels. Sleep was also recorded continuously over the same period. MEASUREMENTS AND RESULTS: Anandamide increased adenosine levels compared to vehicle controls with the peak levels being reached during the third hour after drug injection. There was a significant increase in slow-wave sleep during the third hour. The induction in sleep and the rise in adenosine were blocked by the CB1-receptor antagonist, SR141716A. CONCLUSIONS: Anandamide increased adenosine levels in the basal forebrain and also increased sleep. The soporific effects of anandamide were mediated by the CB1 receptor, since the effects were blocked by the CB1 receptor antagonist. These findings identify a potential therapeutic use of endocannabinoids to induce sleep in conditions where sleep may be severely attenuated. PMID- 14746373 TI - Liver temperature during sleep. AB - STUDY OBJECTIVE: The liver, like brown adipose tissue, is an important source of nonshivering thermogenesis. We tested the hypothesis that the thermal state of the liver is preserved during slow wave sleep but deteriorates during paradoxical sleep. METHODS: Twelve adult male Wistar rats were equipped with electrodes and thermistor probes--which measured cortical (Tco), interscapular brown adipose tissue (Tibat), and liver (Tl) temperatures--for thermal and sleep studies. Six rats were exposed to thermoneutral (24 degrees C) and 6 to low (9 degrees C) ambient temperature. Isolated paradoxical sleep episodes preceded and followed by more than 3 minutes of wakefulness or slow wave sleep were analyzed. The Tco, Tibat, and Tl levels were also assessed during the slow wave sleep that preceded each isolated paradoxical sleep episode. RESULTS: During slow wave sleep, the Tibat and Tl were clearly higher in rats exposed to the cold than in those exposed to the thermoneutral condition (< 1 degrees C). During paradoxical sleep, however, these 2 temperatures decreased in rats exposed to the cold condition, and the changes observed for Tco were completely inversed. CONCLUSIONS: These results support the hypothesis that the thermal state in the liver of rats deteriorates during paradoxical sleep and suggest that nonshivering thermogenesis in the liver contributes to the defense of global thermal homeostasis in the sleeping endothermic organism, especially during slow wave sleep. PMID- 14746374 TI - Effects of IV and ICV hypocretin-1 (orexin A) in hypocretin receptor-2 gene mutated narcoleptic dogs and IV hypocretin-1 replacement therapy in a hypocretin ligand-deficient narcoleptic dog. AB - STUDY OBJECTIVES: Using two different canine models of narcolepsy, we evaluated the therapeutic effects of hypocretin-1 on cataplexy and sleep. MEASUREMENTS AND RESULTS: Intracerebroventricular administration of hypocretin-1 (10 and 30 nmol per dog) but not intravenous administration (up to 6 microg/kg) induced significant wakefulness in control dogs. However, hypocretin-1 had no effect on cataplexy or wakefulness in hypocretin receptor-2 gene (Hcrtr2) mutated narcoleptic Dobermans. Only very high intravenously doses of hypocretin-1 (96-384 microg/kg) penetrated the brain, to produce a short-lasting anticataplectic effect in a hypocretin-ligand-deficient animal. CONCLUSIONS: Hypocretin-1 administration, by central and systemic routes, does not improve narcoleptic symptoms in Hcrtr2 mutated Dobermans. Systemic hypocretin-1 hardly crosses the blood-brain barrier to produce therapeutic effects. The development of more centrally penetrable and longer lasting hypocretin analogs will be needed to further explore this therapeutic pathway in humans. PMID- 14746375 TI - Hypocapnia decreases the amount of rapid eye movement sleep in cats. AB - CONTEXT: Sleep is disturbed at high altitudes. Low PO2 levels at high altitude cause hyperventilation, which results in secondary hypocapnia (low PaCO2 levels). Thus, although sleep disruption at high altitudes is generally assumed to be caused by hypoxia, it may instead be the result of hypocapnia. OBJECTIVE: To determine whether hypocapnia disrupts sleep. METHODS: Four cats were studied for a total of 345 hours of sleep recordings. Two methods were used to test this idea. First we studied their sleep when the cats breathed oxygen concentrations (15% and 10%) equivalent to those at approximately 12,000 feet and 21,000 feet. Then we studied their sleep again in response to the same hypoxic stimuli but with CO2 added to the inspirate to maintain normal CO2 levels. Second, we used mechanical hyperventilation to vary the levels of CO2 while maintaining normal O2 levels. RESULTS: Hypoxia (10% O2) decreased the amount of rapid eye movement sleep to about 20% of normal, and adding back CO2 restored rapid eye movement sleep to approximately 70% of normal. Periodic breathing and apneas were not observed during hypoxia in sleep. When mechanical hyperventilation lowered the CO2 to 85%, 75%, and 65% of normal, rapid eye movement sleep decreased progressively from a control level of 17% of total recording time to 12%, 7%, and 4%, respectively. CONCLUSION: We conclude that hypocapnia rather than hypoxia may account for most of the sleep disturbance at high altitudes. PMID- 14746376 TI - Electroencephalographic activities during wakefulness and sleep in the frontal cortex of healthy older people: links with "thinking". AB - STUDY OBJECTIVE: Separate studies have shown for the frontal cortex that: i) sleep electroencephalogram (EEG) activity less than 1 Hz may be linked to waking recovery, and ii) waking frontal theta activity may reflect "cortical workload." We explored the potential linkage between (i) and (ii) in relation to specific waking theta frequencies and 0.5-Hz to 1.0-Hz activity in sleep. DESIGN: A correlational study SETTING: Laboratory-based waking EEGs under different (eyes closed) contrived "thinking" conditions (mostly localized to the left frontal area), and night sleep EEGs at home. PARTICIPANTS: Twelve right-handed, healthy, good-sleeping, older adults (mean age, 67.3 years) MEASUREMENT & RESULTS: EEGs comprised 4 bipolar derivations (Fp1-F3; Fp2-F4; O1-P3; O2-P4). Power in the 7-8 Hz bin was the only waking frequency significantly (positively) correlated with power in the 0.5- to 1.0-Hz bin during the first NREM period and only for the left frontal EEG. Interestingly, 7- to 8-Hz power immediately after lights out at bedtime, and before the appearance of EEG sleepiness, showed an even higher positive correlation with 0.5- to 1.0-Hz power during sleep. This wake-sleep EEG link was confirmed in another sample of 8 similar participants. Waking frontal 7- to 8-Hz EEG may not be typical theta, but "kappa" activity, believed to be associated with thinking CONCLUSIONS: Within limits of the EEG, and for the left frontal area, waking thinking may be reflected by putative cortical reorganization during the first NREM period. PMID- 14746377 TI - Rapid eye movement density is reduced in the normal elderly. AB - STUDY OBJECTIVES: While there is general agreement on the age changes in non rapid eye movement sleep, there is conflicting evidence on whether eye movement density (EMD) in rapid eye movement sleep is affected by aging. We therefore performed computer measurement of EMD in young and elderly normal subjects. DESIGN: Sleep electroencephalogram and electrooculogram were recorded in each subject on 4 nonconsecutive baseline nights. Eye movement density in the elderly subjects was compared to that in young adults. SETTING: A sleep research laboratory with 4 separate bedrooms. INTERVENTIONS: Not applicable. PARTICIPANTS: Subjects were 19 young normal adults and 19 elderly normal adults. MEASUREMENTS AND RESULTS: Digitized electrooculograms were analyzed with the extensively validated zero-cross period-amplitude module of PASS PLUS software. The EMD was measured as 0.3 to 2 Hz integrated amplitude per 20-second stage of rapid eye movement sleep. Eye-movement incidence was the number of half waves. Eye-movement amplitude was the sum of peak-trough excursions (curve length) in the average half wave. We also counted visually the number of 2-second epochs with eye movements for 1 baseline night in both groups. The EMD in the elderly subjects was substantially and significantly lower than in the young subjects. Visual scoring of EMD on 1 baseline night confirmed the statistically significant difference between age groups. Period-amplitude analysis revealed that a lower eye-movement incidence rather than reduced amplitude caused the lower EMD in the elderly. The EMD was significantly correlated within subjects across the nonconsecutive baseline nights in both groups; in both, subjects' EMD average across 2 nights provided a correlation greater than .90 with the 4-night mean. CONCLUSIONS: The incidence of eye movements during rapid eye movement sleep is substantially reduced in the elderly. We hypothesize that this reduction is due to degenerative (aging) rather than developmental brain changes. The correlation analysis indicates that EMD is a reasonably stable individual trait in both young and elderly adults. These results encourage normative studies of EMD over a wider age span and continued exploration of the relation of EMD to cognitive function in the elderly. PMID- 14746378 TI - Ethanol and sleep loss: a "dose" comparison of impairing effects. AB - STUDY OBJECTIVES: Studies to assess the risks associated with sleep loss relative to the well-documented risks of alcohol are limited in number and design. This study compared the "dose"-related sedative, performance-impairing, and amnestic effects of sleep loss to those of ethanol ingestion. DESIGN: Mixed-design experiment with random assignment to a sleep loss (n=12) or ethanol (n=20) group, with each participant assessed under 4 conditions. PARTICIPANTS: Thirty-two healthy normal adult volunteers, aged 21 to 35 years. INTERVENTIONS: In sleep loss, participants had 8, 6, 4, and 0 hours time in bed, producing 0, 2, 4, and 8 hours of sleep loss. For ethanol, participants ingested 0.0 g/kg, 0.3 g/kg, 0.6 g/kg, and 0.9 g/kg ethanol from 8:30 AM to 9:00 AM after 8 hours of time in bed the previous night. Each participant received his or her 4 doses of ethanol or sleep loss in a Latin square design with 3 to 7 days between doses. MEASUREMENTS: All subjects completed the Multiple Sleep Latency Test (MSLT) at 9:30 AM, 11:30 AM, 1:30 PM, 3:30 PM, and 5:30 PM and a performance battery at 10:00 AM, 12:00 NOON, 2:00 PM, and 4:00 PM consisting of memory, psychomotor vigilance, and divided attention tests. RESULTS: Ethanol and sleep loss reduced the average daily sleep latency on the MSLT, both as a linear function of dose, with sleep loss in hours being 2.7 times more potent than ethanol in grams per kilogram. Ethanol and sleep loss also slowed reaction time on the psychomotor vigilance test in a linear dose-related function with the 2 being equipotent in their impairing effect. On the divided attention test, tracking deviations were increased by both ethanol and sleep loss in an equipotent and linear dose-related function. Memory recall was reduced in a linear dose-related function by both ethanol and sleep loss with ethanol being slightly more potent. Finally, sleep loss doses produced a linear decrease in self-rated quality of performance, while only at the highest ethanol dose was performance rated as poorer. CONCLUSIONS: At the studied doses, sleep loss was more potent than ethanol in its sedative effects but comparable in effects on psychomotor performance. Ethanol produced greater memory deficits, and subjects were less aware of their overall performance impairment. PMID- 14746379 TI - Total sleep deprivation elevates blood pressure through arterial baroreflex resetting: a study with microneurographic technique. AB - STUDY OBJECTIVES: Sleep deprivation has a profound effect on cardiovascular regulation through the autonomic nervous system. This study examined the effect of 24-hour total sleep deprivation on muscle sympathetic nerve activity (MSNA), which is a direct measurement of the postganglionic sympathetic efferent innervating the vascular bed in the skeletal muscle and other circulatory structures. DESIGN: The study was performed on 6 young healthy men. The factors exerting influence on MSNA, such as aging, obesity, body posture, activity, intensity of illumination, and food and beverage consumption were strictly controlled. Burst rate and burst incidence were used as parameters of MSNA. The burst rate, burst incidence, heart rate, and systolic and diastolic blood pressure were measured after total sleep deprivation and control sleep. To perform a linear regression analysis of arterial baroreflex (ABR), the incidence of MSNA bursts corresponding to a given diastolic blood pressure (%MSNA) was examined. MEASUREMENT AND RESULTS: The diastolic blood pressure was significantly higher after total sleep deprivation than after control sleep (66.5 +/- 1.7 vs 57.4 +/- 3.3 mm Hg). The burst rate (9.6 +/- 1.8 vs 13.3 +/- 2.7 bursts/min) and burst incidence (21.6 +/- 4.5 vs 30.3 +/- 8.9 bursts/100 heart beats) of MSNA were significantly lower after total sleep deprivation than after control sleep (P < .05). Analysis of the ABR disclosed a significant linear regressive relation between %MSNA and diastolic blood pressure in every subject after both total sleep deprivation and control sleep. This result implies that the ABR regulates the occurrence of MSNA bursts under different diastolic blood pressure conditions. The threshold (X-axis intercept) of the blood pressure regression line (ie, an indicator of the ABR set point) shifted by 12 +/- 4.3 mm Hg toward a higher blood pressure level after total sleep deprivation (P < .05). The ABR sensitivity, or the slope of the regression line, tended to be less steep after total sleep deprivation than after control sleep, although it was not statistically significant (P = .09). CONCLUSIONS: The diastolic blood pressure increased and both burst rate and burst incidence of MSNA decreased after total sleep deprivation. The results show that resetting of the ABR toward a higher blood pressure level occurred after total sleep deprivation. This ABR resetting probably brings about an increase in arterial blood pressure after total sleep deprivation. PMID- 14746380 TI - Instrumental REM sleep deprivation in neonates leads to adult depression-like behaviors in rats. AB - STUDY OBJECTIVES: Previous studies have demonstrated that neonatal suppression of rapid eye movement (REM) sleep by pharmacologic agents, particularly clomipramine, produces adult depressive behavior. These findings suggest the hypothesis that REM sleep deprivation (RSD) mediates the depressogenic behaviors of neonatally administered antidepressant drugs. Drug suppression of RSD, however, was thought to be confounded by the other effects of the drugs. The current study was aimed to show the adult effect of neonatal RSD in rats by instrumental means, ie, a computer-controlled shaking method. DESIGN: Three treatment groups were studied: an instrumental RSD group, a yoked control group, and a nonshaken, maternally separated, control group. All treatments began at the age of 14 days and lasted for 7 days. Adult behavior measurements including tests of sexual activity, locomotor activity, shock-induced fighting, and sleep recording were subsequently performed. MEASUREMENTS AND RESULTS: The major findings of our investigation were that rats subjected to neonatal instrumental RSD demonstrated diminished sexual activity, decreased aggressive behavior, increased percentage of REM sleep, and decreased wake-REM sleep ratio compared with yoked control rats. These data are compatible with the findings from adult rats subjected to neonatal treatment with the REM-sleep suppressant, clomipramine, and supports the hypothesis that neonatal RSD results in adult depressive abnormalities. CONCLUSION: Neonatal RSD induced by a nondrug method results in adult depression-like changes similar to those induced by a REM-sleep suppressant drug, although the extent of these changes varies. PMID- 14746381 TI - Adenotonsillectomy improves neurocognitive function in children with obstructive sleep apnea syndrome. AB - OBJECTIVE: To evaluate neurocognitive functions of children with obstructive sleep apnea syndrome (OSAS), before and after adenotonsillectomy, compared with healthy controls. DESIGN: Prospective study. PATIENTS AND METHODS: Thirty-nine children with OSAS aged 5 to 9 years (mean age, 6.8 +/- 0.2 years) and 20 healthy children (mean age, 7.4 +/- 1.4 years) who served as controls, underwent a battery of neurocognitive tests containing process-oriented intelligence scales. Twenty-seven children in the OSAS group underwent follow-up neurocognitive testing 6 to 10 months after adenotonsillectomy. Fourteen children in the control group were also reevaluated 6 to 10 months after the first evaluation. RESULTS: Children with OSAS had lower scores compared with healthy children in some Kaufman Assessment Battery for Children (K-ABC) subtests and in the general scale Mental Processing Composite, indicating impaired neurocognitive function. No correlation was found between neurocognitive performance and OSAS severity. Six to 10 months after adenotonsillectomy, the children with OSAS demonstrated significant improvement in sleep characteristics, as well as in daytime behavior. Their neurocognitive performance improved considerably, reaching the level of the control group in the subtests Gestalt Closure, Triangles, Word Order, and the Matrix analogies, as well as in the K-ABC general scales, Sequential and Simultaneous Processing scales, and the Mental Processing Composite scale. The magnitude of the change expressed as effect sizes showed medium and large improvements in all 3 general scales of the K-ABC tests. CONCLUSIONS: Neurocognitive function is impaired in otherwise healthy children with OSAS. Most functions improve to the level of the control group, indicating that the impaired neurocognitive functions are mostly reversible, at least 3 to 10 months following adenotonsillectomy. PMID- 14746382 TI - Prevalence of sleep-disordered breathing in children with Down syndrome: polygraphic findings in 108 children. AB - STUDY OBJECTIVES: To assess the prevalence of sleep-disordered breathing in a nonselected group of children with Down syndrome and to determine significant predisposing factors for this condition. DESIGN: Prospective study. SETTING: Tertiary care university hospital in Madrid, Spain. PATIENTS: The study population included 108 consecutive children with Down syndrome (mean [SD] age, 7.9 [4.5] years; range, 1-18 years) independently of whether or not suggestive clinical features of sleep-disordered breathing were present. INTERVENTIONS: In addition to history, physical examination, and lateral radiographs of the nasopharynx, all participants underwent an overnight cardiorespiratory polygraphy at the hospital using a portable ambulatory device (Apnoescreen II plus). An apnea-hypopnea index of at least 3 was required for defining the presence of sleep-disordered breathing. RESULTS: The prevalence of sleep-disordered breathing was 54.6%, with a significantly higher prevalence in boys (64.7%) than in girls (38.5%) (P < .05). The group with sleep-disordered breathing was significantly younger (6.4 [3.9] years) than those with normal polysomnographic recordings (9.6 [4.6] years) (P < .001). In the multivariate analysis, age (less than 8 years old) (odds ratio [OR], 3.36; 95% confidence interval [CI], 1.40, 8.06); male sex (OR, 3.32; 95% CI, 1.32, 8.12); and tonsillar hyperplasia (OR, 5.24; 95% CI, 1.52, 19.03) were significantly associated with sleep-disordered breathing. Body mass index, adenoid hyperplasia, previous tonsillectomy or adenoidectomy, congenital heart disease, malocclusion, and macroglossia did not affect the prevalence of sleep-disordered breathing. CONCLUSIONS: The prevalence of sleep disordered breathing in children with Down syndrome is very high, particularly in boys. Tonsillar hyperplasia may play a role in the pathophysiology of sleep disordered breathing in these patients. Adenoid hyperplasia, obesity, and congenital heart disease were not important risk factors for sleep-disordered breathing. PMID- 14746383 TI - Improving pediatric compliance with positive airway pressure therapy: the impact of behavioral intervention. AB - STUDY OBJECTIVE: The purposes of this study were (1) to describe behavior interventions provided to improve compliance with positive airway pressure (PAP) therapies in children with obstructive sleep apnea, (2) to survey physician and caregiver satisfaction with these interventions, and (3) to present preliminary outcome data on the effects of these interventions. DESIGN: Retrospective, descriptive analysis. SETTING: Pediatric psychology consultation service at a university-affiliated rehabilitation hospital. PARTICIPANTS: 20 children (aged 1 17 years) with obstructive sleep apnea, referred by physicians for noncompliance with PAP. INTERVENTIONS: Patients self-selected into 1 of 3 groups: (1) a group receiving a 1.5-hour consultation and recommendation session (CR+), (2) a group receiving consultation and recommendations plus a course of behavior therapy (BT), and (3) a group for whom behavior therapy was recommended after the consultation and recommendations, but the family did not follow-up (CR-). RESULTS: Prior to behavior intervention, none of the children were consistently wearing the PAP equipment. After intervention, 75% of children who received behavior intervention (CR+ and BT groups) successfully tolerated PAP with increased hours of documented usage. This was in contrast to children whose families declined recommended behavior therapy (CR- group), of whom 0% increased their usage of PAP. High satisfaction ratings were obtained from referring physicians and patient caregivers for children in the CR+ and BT groups. CONCLUSIONS: The results are encouraging and support the importance of behavior analysis and therapy for increasing compliance and making the benefits of PAP available to a greater number of children. PMID- 14746384 TI - Willingness to pay for polysomnography in children with obstructive sleep apnea syndrome: a cost-benefit analysis. AB - OBJECTIVES: To analyze willingness to pay (WTP) for polysomnography (PSG) among parents of children with obstructive sleep apnea syndrome (OSAS). To analyze the cost-benefit of PSG in a collectively funded healthcare system. SETTING: University-affiliated sleep laboratory. SUBJECTS: Parents of 158 boys and 94 girls, who had a mean age of 6.0 +/- 3.9 years. The telephone survey, using a contingent valuation approach, was conducted with 3 groups of parents: those whose children were scheduled for PSG (n = 83), whose children were had had PSG within the previous 6 months (n = 77), and whose children had had PSG and adenotonsillectomy in the previous 6 months (n = 92). RESULTS: Two hundred and fifty-two parents (92% compliance rate), 75% of whom were mothers, responded to the WTP interview. Multivariate analysis revealed that the independent variables influencing WTP were bid (OR = 0.745, P < .001), age times bid (OR = 0.835, P < .05), and affected health status (OR = 3.5, P < .001). The median WTP value for PSG studies of children with OSAS following adenotonsillectomy was dollars 762 plus the savings of dollars 60 to the health care system-subtracting the cost of the dollars 250 PSG study resulted in a monetary benefit of dollars 572 per diagnosis. CONCLUSIONS: We conclude that PSG diagnosis for children with OSAS is beneficial. Decision makers and sleep specialists can use WTP to prioritize allocation of resources to increase the availability of PSG studies for children. PMID- 14746385 TI - Tolerance of esophageal pressure monitoring during polysomnography in children. AB - OBJECTIVES: To assess tolerance of esophageal pressure monitoring (EPM) among 5- to 13-year-old children during research polysomnography at study entry and again 1 year later. DESIGN: Prospective, observational study. SETTING: University-based sleep laboratory. PARTICIPANTS: Children scheduled for adenotonsillectomy or hernia repair. INTERVENTIONS: None; all operations were performed for clinical indications only. RESULTS: Forty-two of 336 families approached about the study declined to participate mainly to avoid EPM. The EPM was usually the main concern for the 47 adenotonsillectomy and 7 hernia-repair patients and families who did participate. Among 54 enrolled subjects, 51 allowed attempts at insertion of the esophageal catheter, and insertion was successful in all cases; 38 tolerated EPM for at least 2 hours; 33 maintained EPM for the entire night; and 36 had repeat EPM 1 year later for at least 2 hours. Reasons for EPM failure included crying at insertion, vomiting, pain, and inadvertent catheter removal during sleep. The children who tolerated EPM for at least 2 hours did not differ from other subjects based on age, sex, presence of a disruptive behavior disorder, anxiety, tonsil size, history of tonsillitis, or body mass index (all P > .05). CONCLUSIONS: The EPM was well tolerated in most school-aged volunteers, but many families did not volunteer, and some children were not able to endure EPM for at least 2 hours. Although better success might be achieved in clinical settings if EPM is medically indicated and not part of voluntary research, EPM is still likely to create significant concern, for children and parents, that must be weighed against anticipated benefits. PMID- 14746386 TI - Situational insomnia: consistency, predictors, and outcomes. AB - STUDY OBJECTIVES: The purpose of this study was to determine the consistency of situational insomnia across several stressful conditions, including the first night in the sleep laboratory, phase advance of sleep time by 3 hours, phase advance of sleep time by 6 hours, and administration of 400 mg of caffeine. The impact of situational insomnia on alertness, metabolic rate, and cardiac measures on the following day was also measured. DESIGN AND SETTING: Subjects spent 5 to 7 nights and the following days in the laboratory. Standard polysomnographic recordings were made on each night. On each day, subjects had a Multiple Sleep Latency Test, performance testing, and metabolic and heart-rate observation periods. PARTICIPANTS: Fifty adult normal sleepers. INTERVENTIONS: Subjects had 1 night with their sleep phase advanced by 3 hours, 1 night with sleep phase advanced by 6 hours, and 1 night with the administration of 400 mg of caffeine 30 minutes prior to lights out. MEASUREMENT AND RESULTS: Sleep efficiency was reduced and variability was increased in each of the stressful conditions, as predicted. Those subjects with the greatest sleep efficiency on the adaptation night (top 25%) were compared with those subjects with the lowest sleep efficiency on the adaptation night (bottom 25%). Those subjects with the poorest sleep on the adaptation night (situational insomnia) had normal sleep on the baseline night that followed but had significantly reduced sleep efficiency when their sleep was advanced or they were given caffeine. Those same subjects had a significant decrease on their Multiple Sleep Latency Test on the day following the 6-hour advance and a significant increase in their Multiple Sleep Latency Test on the day following caffeine administration. The good sleepers had no significant change in their Multiple Sleep Latency Test during any of the study conditions. In terms of demographic variables, the situational insomnia group used less alcohol and tended to include a higher percentage of men. The situational insomnia group also had an elevated heart rate and increased low frequency and decreased high-frequency electrocardiographic spectral power compared to the good sleepers. Significant differences were not found on personality or historical reports of poor sleep. CONCLUSIONS: Normal young adults have a consistent response to various types of situational stress. Those individuals who respond with poor sleep may display increased sleepiness associated with their poor sleep but may also be more sensitive to the effects of caffeine. These individuals have cardiac changes consistent with sympathetic nervous system activation, and they may be at risk for developing insomnia and other associated disorders. PMID- 14746387 TI - Does sleep deprivation worsen mild obstructive sleep apnea? AB - STUDY OBJECTIVES: Sleep deprivation is believed to worsen obstructive sleep apnea (OSA). We assessed the effect of acute sleep deprivation on polysomnography in a cohort of subjects with mild OSA and a cohort of subjects without OSA. DESIGN: Crossover study in which subjects initially had polysomnography after a normal night's sleep or after 36 hours of sleep deprivation, followed by a 2- to 4-week interval, after which subjects were restudied under the alternate testing condition. SETTING AND PARTICIPANTS: 13 subjects with mild OSA and 16 subjects without OSA were studied in a university teaching hospital sleep laboratory. INTERVENTIONS: 36 hours of supervised sleep deprivation. MEASUREMENTS: Subjects' age, body mass index, neck circumference and Epworth Sleepiness Scale scores were measured; actigraphy and sleep diaries were used to estimate prior sleep debt before each sleep study. RESULTS: Sleep deprivation was found to significantly increase total sleep time, sleep efficiency, and rapid eye movement and slow-wave sleep time. Subjects with OSA showed a lower minimum oxygen saturation after sleep deprivation. However, subjects did not show a significantly different respiratory disturbance index, arousal index, or length of the longest apnea after sleep deprivation. CONCLUSIONS: Acute sleep deprivation did not worsen most OSA parameters as measured by polysomnography. A lower minimum oxygen saturation in mild OSA subjects after sleep deprivation may be important in patients with significant cardiorespiratory disease. More research is needed to assess whether daytime performance and function (eg, driving, sleepiness) is more greatly impaired in OSA subjects who are sleep deprived, compared to normal subjects who are sleep deprived. PMID- 14746388 TI - Arousals in sleep-disordered breathing: patterns and implications. AB - STUDY OBJECTIVE: To describe the whole spectrum of electroencephalographic (EEG) transients associated with the termination and recovery of obstructed respiratory events and, thus, widen the recognized spectrum of arousal phenomena from sleep. DESIGN: Retrospective review of diagnostic polysomnograms. SETTING: American Academy of Sleep Medicine (AASM)-accredited multidisciplinary sleep disorders center. PATIENTS: 17 patents with obstructive sleep-disordered breathing. INTERVENTIONS: None. MEASUREMENTS AND RESULTS: Nasal airflow using a nasal cannula-pressure-transducer system and oral flow by a thermistor were used to score apneas and hypopneas; the latter included flow-limitation events. The EEG patterns that crested or occurred within 2 to 3 seconds of respiratory recovery were recorded, and posthoc categories were created for the purpose of tabulation ranging from an AASM 3-second arousal to a single K-complex with no electromyographic increase. Chi-square statistic was calculated to assess the difference in EEG patterns at event termination between apneas and hypopneas. Score-rescore agreement was tested. Apneas were significantly more likely to be associated with a 3-second arousal than were hypopneas, but all types of EEG change were seen with both types of events. Spindles were rarely seen with arousal-linked K-complexes. The majority of events in rapid eye movement sleep were terminated with visible electromyography tone increase. CONCLUSIONS: The spectrum of EEG change associated with the termination of respiratory events identified by using a nasal-cannula-pressure-transducer system is wider than that recognized as arousal phenomena by the 1992 AASM criteria. Scoring arousals with the 3-second rule may falsely minimize the apparent impact of abnormal breathing on sleep. The time may be right to update arousal recognition rules. PMID- 14746389 TI - Assessment of sleep and sleepiness in Parkinson disease. AB - STUDY OBJECTIVES: To develop a short and practical scale (SCOPA-SLEEP) that evaluates nighttime sleep and daytime sleepiness. The scale is developed for research in Parkinson disease but may be of value for other somatic diseases. DESIGN: Postal survey including 4 instruments, the SCOPA-SLEEP nighttime sleep (5 items) and daytime sleepiness (6 items), the Pittsburgh Sleep Quality Index, and the Epworth Sleepiness Scale. SETTING: Movement Disorders Center, Department of Neurology, Leiden University Medical Center, Leiden, The Netherlands. PARTICIPANTS: 143 patients with Parkinson disease and 104 controls. INTERVENTIONS: N/A. MEASUREMENTS AND RESULTS: Reliability of the scale was high: internal consistency of the nighttime sleep and daytime sleepiness scales were 0.88 and 0.91, respectively (Cronbach alpha), and test-retest reliabilities were 0.94 and 0.89, respectively (intraclass correlation coefficient). Scale scores differed significantly between patients and controls (P < .001). Construct validity was assessed by correlations with scales that addressed similar constructs. Correlation between the nighttime sleep scale and the Pittsburgh Sleep Quality Index was 0.83 (P < .001), and the correlation between the daytime sleepiness scale and the Epworth Sleepiness Scale was 0.81 (P < .001). Factor analysis revealed 1 factor each for both scales, indicating that the scales measure 1 construct, which justifies the calculation of sumscores. The coefficient of variation of both the nighttime sleep and the daytime sleepiness scale was higher than that of the Pittsburgh Sleep Quality Index and the Epworth Sleepiness Scale, indicating a better ability to detect differences between individuals. CONCLUSIONS: The SCOPA-SLEEP is a reliable and valid instrument for assessing nighttime sleep and daytime sleepiness in patients with Parkinson disease. PMID- 14746390 TI - Analysis of CAG repeat expansions in restless legs syndrome. AB - STUDY OBJECTIVES: A relatively high prevalence of restless legs syndrome symptoms has been recently reported in a substantial proportion of patients affected with spinocerebellar ataxia type 3. Our aim was to investigate whether there is a common genetic etiology between restless legs syndrome and spinocerebellar ataxia type 3. DESIGN: Systematic differences in the number of spinocerebellar ataxia type 3 trinucleotide repeat were investigated by means of an association study. The relationship between the size of the expanded alleles and several clinical features was also considered. PARTICIPANTS AND SETTING: 125 extensively characterized restless legs syndrome patients compared with 188 healthy controls matched for ethnic background. INTERVENTIONS: N/A. MEASUREMENTS AND RESULTS: No CAG repeat expansions in the pathologic or intermediate range were detected in any of the examined subjects, including patients and controls. A similar allelic distribution was observed in both groups (Mann-Whitney U test = 78406; P = 0.99). Moreover, stratification analyses of the patients' samples according to different clinical and polysomnographic variables disclosed no significant differences. CONCLUSIONS: These results do not provide evidence toward an involvement of large CAG trinucleotide expansions at the spinocerebellar ataxia type 3 locus in idiopathic restless legs syndrome. PMID- 14746391 TI - Potential action of melatonin in insomnia. PMID- 14746393 TI - Increased percentage of slow-wave sleep in REM sleep behavior disorder (RBD): a reanalysis of previously published data from a controlled study of RBD reported in SLEEP. PMID- 14746392 TI - Upper airway management of the adult patient with obstructive sleep apnea in the perioperative period--avoiding complications. AB - PURPOSE: To help practitioners avoid adverse perioperative events in patients with obstructive sleep-disordered breathing. REVIEWERS: Members of the American Academy of Sleep Medicine's Clinical Practice Review Committee. METHODS: A search of MEDLINE database using MeSH terms apnea, obstructive sleep apnea and anesthesia was conducted in October 2001. This review focuses on articles published in English between 1985 and 2001 that pertain to non-upper airway surgery in obstructive sleep apnea patients. RESULTS AND CONCLUSIONS: Scientific literature regarding the perioperative risk and best management techniques for OSAHS patients is scanty and of limited quality. There is insufficient information to develop an AASM standards of practice recommendation. Therefore, the Clinical Practice Review Committee (CPRC) used the available data to make this statement based upon a consensus of clinical experience and published peer reviewed medical evidence. Important components of the perioperative management of OSAHS patients include a high degree of clinical suspicion, control of the airway throughout the perioperative period, judicious use of medications, and appropriate monitoring. Further research is needed to define the magnitude of risk and optimal perioperative care. PMID- 14746394 TI - Normal hypocretin-1 levels in the cerebrospinal fluid of patients with fatal familial insomnia. PMID- 14746395 TI - Creative approach to financing a PACS. PMID- 14746396 TI - Angiogenesis model for ultrasound contrast research: exploratory study. AB - RATIONALE AND OBJECTIVES: To optimize an angiogenesis model for imaging research that is stable and can be imaged several times over the angiogenic time course. MATERIALS AND METHODS: Mice and rats received two injections of 0.4 mL of extract of basement membrane matrix (Matrigel; Becton Dickinson Labware, Bedford, MA) in the subcutaneous spaces on either side of the spine. One of the two Matrigel plugs in each animal had either 0.1 microg/mL of basic fibroblast growth factor (bFGF) (11 mice), 1.0 microg/mL of bFGF (12 mice, 5 rats), or 1.0 microg/mL of bFGF and 60 U/mL of heparin (11 mice). Three to 12 days after implantation, animals were imaged before and after the administration of up to four injections of 0.1 mL AF0150. Phase inversion imaging was used on a Siemens Elegra (Siemens ultrasound, Issaquah, WA) equipped with a 13 MHz VFX transducer. Three observers subjectively assessed the pattern of enhancement using a four-point scale. The Matrigel plugs were then removed and two observers graded the angiogenic response on a four-point scale. Ten Matrigel plugs, five with 1.0 microg/mL bFGF and five without, were evaluated histologically following immunohistochemical staining with anti-CD31. RESULTS: The angiogenic response was greater in Matrigel plugs with 1.0 than with 0.1 microg/mL of bFGF. Heparin did not increase the angiogenic response. Vessels were predominantly at the periphery of the plugs with variable central penetration. Plugs appeared anechoic and homogeneous on ultrasound. Contrast enhancement within the plug occurred in 44% of mice with an angiogenic response at or after day 6 and the enhancement increased with the angiogenic response. In the others, peripheral enhancement could not be distinguished from the enhancement of surrounding tissues that were also hyperemic. The thicker rat skin interfered with plug assessment. CONCLUSION: A stable angiogenesis model without the complexity of tumors is described. This model offers the opportunity to image the development and/or inhibition of angiogenesis. Neovasculature in Matrigel was detectable using ultrasound contrast. Quantitative studies correlating the degree of enhancement to microvascular density will be determined in subsequent studies. PMID- 14746398 TI - Sodium magnetic resonance imaging of proteoglycan depletion in an in vivo model of osteoarthritis. AB - RATIONALE AND OBJECTIVES: The aim of the study was to investigate the feasibility of using sodium magnetic resonance imaging (MRI) as a noninvasive quantitative technique for measuring proteoglycan (PG) content in an in vivo porcine model of osteoarthritis (OA). MATERIALS AND METHODS: Biochemical conditions similar to those of OA were created by an intra-articular injection of recombinant porcine interleukin-1beta (IL-1beta) into the knee joint of pigs (n = 6) before performing MRI. The contralateral knee joint was given a saline injection to serve as an internal control. Sodium MRI data were acquired on a 4-T clinical MR scanner and used to compute quantitative sodium and fixed charge density (FCD) maps based on a previously established methodology. In vivo FCD maps were compared with FCD maps obtained using ex vivo patellae harvested from the specimens. The tissue and joint fluid were subjected to histologic and immunohistochemical analyses as independent measurements of IL-1beta activity and PG loss. RESULTS: The average FCD of IL-1beta-treated patellae was measured to be 49% lower than that of saline-treated patellae, indicating a loss of PG content. These results were supported by histologic and immunochemical findings, most notably a reduction in staining for PG and an increase in matrix metalloproteinases in the synovial fluid. CONCLUSION: Sodium MRI can serve as a quantitative method to measure in vivo changes in PG content in an animal model of OA. The use of a noninvasive quantitative in vivo PG measurement technique such as sodium MRI on an animal model would aid greatly in efforts to monitor the efficacy of treatments for OA. Furthermore, these results indicate that early degenerative events could be detected noninvasively in vivo in humans with PG depleting diseases such as OA. PMID- 14746397 TI - Effect of an imaging-based streamlined electronic healthcare process on quality and costs. AB - RATIONALE AND OBJECTIVES: A streamlined process of care supported by technology and imaging may be effective in managing the overall healthcare process and costs. This study examined the effect of an imaging-based electronic process of care on costs and rates of hospitalization, emergency room (ER) visits, specialist diagnostic referrals, and patient satisfaction. MATERIALS AND METHODS: A healthcare process was implemented for an employer group, highlighting improved patient access to primary care plus routine use of imaging and teleconsultation with diagnostic specialists. An electronic infrastructure supported patient access to physicians and communication among healthcare providers. The employer group, a self-insured company, manages a healthcare plan for its employees and their dependents: 4,072 employees were enrolled in the test group, and 7,639 in the control group. Outcome measures for expenses and frequency of hospitalizations, ER visits, traditional specialist referrals, primary care visits, and imaging utilization rates were measured using claims data over 1 year. Homogeneity tests of proportions were performed with a chi-square statistic, mean differences were tested by two-sample t-tests. Patient satisfaction with access to healthcare was gauged using results from an independent firm. RESULTS: Overall per member/per month costs post-implementation were lower in the enrolled population (126 dollars vs 160 dollars), even though occurrence of chronic/expensive diseases was higher in the enrolled group (18.8% vs 12.2%). Lower per member/per month costs were seen for inpatient (33.29 dollars vs 35.59 dollars); specialist referrals (21.36 dollars vs 26.84 dollars); and ER visits (3.68 dollars vs 5.22 dollars). Moreover, the utilization rate for hospital admissions, ER visits, and traditional specialist referrals were significantly lower in the enrolled group, although primary care and imaging utilization were higher. Comparison to similar employer groups showed that the company's costs were lower than national averages (119.24 dollars vs 146.32 dollars), indicating that the observed result was not attributable to normalization effects. Patient satisfaction with access to healthcare ranked in the top 21st percentile. CONCLUSION: A streamlined healthcare process supported by technology resulted in higher patient satisfaction and cost savings despite improved access to primary care and higher utilization of imaging. PMID- 14746399 TI - Artificial neural networks (ANNs) for differential diagnosis of interstitial lung disease: results of a simulation test with actual clinical cases. AB - RATIONALE AND OBJECTIVES: To evaluate the performance of an artificial neural network (ANN) scheme with use of consecutive clinical cases and its effect on radiologists with an observer test. MATERIALS AND METHODS: Artificial neural networks were designed to distinguish among 11 interstitial lung diseases on the basis of 26 inputs (16 radiologic findings, 10 clinical parameters). Chest radiographs of 96 consecutive cases with interstitial lung disease were used. Five radiologists independently rated their radiologic findings on the 96 chest radiographs. Based on their ratings of radiologic findings and clinical parameters obtained from the hospital information system, the output values indicating the likelihood of each of the 11 interstitial lung diseases were determined. Subsequently, 30 cases were selected from these 96 cases for an observer test. Five radiologists marked their confidence levels for diagnosis of 11 possible diseases in each case without and with ANN output. The performance of ANNs and radiologists was evaluated by receiver operating characteristic analysis based on their outputs and on confidence levels, respectively. RESULTS; The average Az value (area under the receiver operating characteristic curve) indicating ANN performance for the 96 consecutive cases was 0.85 +/- 0.03. The average Az values indicating radiologists' performance without and with ANN outputs were 0.81 +/- 0.11 and 0.87 +/- 0.06, respectively. The diagnostic accuracy was improved significantly when radiologists read chest radiographs with ANN outputs (P < .05). CONCLUSION: Artificial neural networks for differential diagnosis of interstitial lung disease may be useful in clinical situations, and radiologists may be able to utilize the ANN output to their advantage in the differential diagnosis of interstitial lung disease on chest radiographs. PMID- 14746400 TI - Increased manganese concentration in the liver after oral intake. AB - RATIONALE AND OBJECTIVES: Manganese is a well-known liver-specific agent used in magnetic resonance imaging. For this purpose, manganese is now administered intravenously. In theory it should be possible to increase the gastrointestinal uptake of manganese through the use of nutritional products as promoters. Such an agent has now been formulated. As part of a primary pharmacologic investigation, the uptake of manganese in the kidney, heart, and liver was studied. MATERIALS AND METHODS: One hundred two female Sprague Dawley rats fasted for 18 hours before the agent (CMC-001) was given orally by gavage. One hundred micromol/kg BW was given to all rats except six, who served as controls. Various concentrations of the promoters (vitamin D3 and the amino acid alanine) were also given. Three hours after administration the rats were killed and the heart, the liver and kidneys were removed. The manganese content was determined by atomic absorption. RESULTS: No systematic increased concentration of manganese was found in either the kidneys or the heart; whereas the manganese content of the liver (approximately 100%) increased significantly compared with the controls and the group receiving pure manganese. No side effects were observed. CONCLUSION: It is possible to increase the gastrointestinal uptake of manganese in fasting rats and thereby increase the concentration in the liver. PMID- 14746401 TI - Qualitative diagnosis of calvarial metastasis by neural network and logistic regression. AB - RATIONALE AND OBJECTIVES: To simplify the diagnostic features used by an artificial neural network compared with logistic regression (LR) in the diagnosis of calvarial metastasis with computed tomography and analyze their accuracy. MATERIALS AND METHODS: Twenty-one of 167 patients with calvarial lesions were found to have metastasis. Clinical and computed tomography data were used for LR and neural network models. Both models were tested with the leave-one-out method. The final results of each model were compared using the area under receiver operating characteristic curve (Az). RESULTS: The neural network identified metastasis significantly more successfully than LR with an Az of 0.9324 +/- 0.0386 versus 0.9192 +/- 0.0373, P = .01. The most important features selected by the LR and neural network were age and edge definition. CONCLUSION: Neural networks offer wide possibilities over statistics for the study of calvarial metastases other than their minimum clinical and radiologic features for diagnosis. PMID- 14746402 TI - Differentiation of cysts from solid tumors in the breast with diffuse optical tomography. AB - RATIONALE AND OBJECTIVES: Near-infrared diffuse optical tomography (DOT) is an emerging imaging technology that has the potential to offer enhanced contrast resolution over the existing technologies for detection and diagnosis of breast cancer. Thus far, the clinical evaluation of DOT has been largely limited to solid tumors. A pilot clinical study focused on DOT imaging of breasts with cysts is presented. MATERIALS AND METHODS: Six cases were studied using the recently developed compact, parallel-detection DOT system. Images characterizing the tissue absorption and scattering were obtained with a finite element-based reconstruction algorithm. The optical images were compared with the mammograms and sonograms. In one case, in vitro measurements of optical properties were conducted for the fluid obtained from needle aspiration. RESULTS: Substantial contrast between cyst and adjacent parenchyma is observed. For the six cases evaluated, the locations and sizes of cysts imaged optically are accurate and consistent with the mammographic and sonographic findings. For the case that aspiration was performed, the absorption and scattering coefficients imaged in the cyst region are quantitatively accurate compared with that measured in vitro from the fluid aspirated. CONCLUSION: This pilot study shows that cysts ranging from 1-4 cm in diameter can be quantitatively imaged. They can be differentiated from solid breast tumors because cysts generally demonstrate lower absorption and scattering coefficients compared with the surrounding normal tissue, whereas solid tumors show concurrent higher absorption and scattering related to the normal tissue. PMID- 14746403 TI - Gadolinium-enhanced computed tomography angiography in multi-detector row computed tomography: initial observations. AB - RATIONALE AND OBJECTIVES: The feasibility of using gadolinium contrast medium for computed tomography angiography (CTA) in multi-detector row computed tomography and the effect of contrast medium dilution was investigated. MATERIALS AND METHODS: Three pigs were each scanned in multiple sessions with injections of non dilute and dilute contrast medium at a dose of 0.3 mmol/kg body weight. Non spiral dynamic scanning at a fixed mid-abdominal aortic level and thoracoabdominal CTA were performed. RESULTS: The magnitude of peak aortic enhancement was not significantly different between dilute and non-dilute contrast medium injections (P = .88), but the former showed earlier enhancement (mean of 2.3 seconds sooner, P < .01) than the latter. CT angiography with gadolinium contrast medium showed much lower enhancement than iodine contrast medium, but small vessels were readily identifiable. CONCLUSION: Gadolinium contrast medium combined with multi-detector row computed tomography may provide clinically useful CTA. Dilution of contrast medium shortens the enhancement time but has little effect on the magnitude. PMID- 14746404 TI - Should pay in radiology be tied to productivity? The case in favor. PMID- 14746405 TI - Reporting instruction for radiology residents. AB - RATIONALE AND OBJECTIVES: To determine the amount of formal instruction and evaluation about reporting given to radiology residents in the U.S.A., to document report generation methods and to quantify the performance of physician coding. MATERIALS AND METHODS: E-mail requests with links to a web-based, anonymous survey were sent to program directors of all accredited radiology residencies in the USA. Demographic questions included university or private affiliation, number of residents, geographic location, and number of hospitals covered. Subject-specific items covered the amount of didactic instruction, formal evaluation of reports, and use of structured reports. A didactic activity index (DAI) was calculated as the sum of answers to domain-specific questions and tested for relation to demographic variables. We also asked about dictation methods and International Classification of Diseases (ICD) or Common Procedural Terminology (CPT) coding of examinations by radiologists. RESULTS: Of the 191 active radiology residencies, 151 (79%) completed the survey. Responses for hours of didactic instruction in reporting given more than a 4-year residency were distributed as follows: 0-1 = 40%, 2-4 = 46%, >4 = 14%. The percentage of resident reports formally graded was distributed as follows: 0-1 = 82%, 2-4 = 8%, >4 = 10%. The extent to which faculty-designed, structured reports were used by residents was distributed as follows: none = 16%, minimal = 25%, few = 17%, some = 33%, most = 9%. The DAI was normally distributed with a mean of 14.8 and a standard deviation of 2.4. Military programs had higher DAIs than university residencies (P = .03). There was no significant relation between any other program demographic variables and the DAI (P > .05). A substantial number of programs reported that physicians performed coding for some or most studies: ICD 9 = 30%, CPT = 26%. The dominant method for report generation was human transcription in 79% followed by speech recognition at 19%. Speech recognition penetration (departments reporting use of the technology for at least some dictation) was estimated to be 38%. CONCLUSION: In 86% of sampled radiology residencies, trainees receive no more than one hour of didactic instruction in radiology reporting per year. An aggregate measure of didactic activity about interpretative reporting was identical across all program demographic variables except that military residencies seemed to do slightly more than those located at universities. PMID- 14746406 TI - How I do it: mentoring the sound-bite generation: part I. The match game. PMID- 14746407 TI - How do I mentor medical students interested in radiology? PMID- 14746408 TI - Financing a large-scale picture archival and communication system. AB - RATIONALE AND OBJECTIVES: An attempt to finance a large-scale multi-hospital picture archival and communication system (PACS) solely based on cost savings from current film operations is reported. MATERIALS AND METHODS: A modified Request for Proposal described the technical requirements, PACS architecture, and performance targets. The Request for Proposal was complemented by a set of desired financial goals-the main one being the ability to use film savings to pay for the implementation and operation of the PACS. RESULTS: Financing of the enterprise-wide PACS was completed through an operating lease agreement including all PACS equipment, implementation, service, and support for an 8-year term, much like a complete outsourcing. Equipment refreshes, both hardware and software, are included. Our agreement also linked the management of the digital imaging operation (PACS) and the traditional film printing, shifting the operational risks of continued printing and costs related to implementation delays to the PACS vendor. An additional optimization step provided the elimination of the negative film budget variances in the beginning of the project when PACS costs tend to be higher than film and film-related expenses. CONCLUSION: An enterprise wide PACS has been adopted to achieve clinical workflow improvements and cost savings. PACS financing was solely based on film savings, which included the entire digital solution (PACS) and any residual film printing. These goals were achieved with simultaneous elimination of any over-budget scenarios providing a non-negative cash flow in each year of an 8-year term. PMID- 14746409 TI - On the repeated use of databases for testing incremental improvement of computer aided detection schemes. PMID- 14746410 TI - Continuous arterial spin-labeling perfusion magnetic resonance imaging of the human testis. AB - RATIONALE AND OBJECTIVES: The purpose of this study was to determine if continuous arterial spin-labeling perfusion magnetic resonance imaging could be used to detect testicular perfusion in human subjects. MATERIALS AND METHODS: Continuous arterial spin-labeling magnetic resonance perfusion imaging was performed in seven normal male volunteers and in one patient with a painful scrotum following vasectomy. RESULTS: Normal testicular blood flow was demonstrated in 14 of 14 normal testes in seven volunteers, as well as in two normally perfused testes in the post-vasectomy patient. A change in the steady state magnetization was observed in all of the normally perfused testes of the seven volunteers. CONCLUSION: It is possible to detect blood flow to the normally perfused testes using noninvasive spin-labeling perfusion magnetic resonance imaging. This modality could potentially, in future investigations, be used to image patients with suspected testicular torsion and resultant testicular ischemia. PMID- 14746411 TI - Motion tracking in MR-guided liver therapy by using navigator echoes and projection profile matching. AB - RATIONALE AND OBJECTIVES: Image registration in magnetic resonance (MR) image guided liver therapy enhances surgical guidance by fusing preoperative multimodality images with intraoperative images, or by fusing intramodality images to correlate serial intraoperative images to monitor the effect of therapy. The objective of this paper is to describe the application of navigator echo and projection profile matching to fast two-dimensional image registration for MR-guided liver therapy. MATERIALS AND METHODS: We obtain navigator echoes along the read-out and phase-encoding directions by using modified gradient echo imaging. This registration is made possible by masking out the liver profile from the image and performing profile matching with cross-correlation or mutual information as similarity measures. The set of experiments include a phantom study with a 2.0-T experimental MR scanner, and a volunteer and a clinical study with a 0.5-T open-configuration MR scanner, and these evaluate the accuracy and effectiveness of this method for liver therapy. RESULTS: Both the phantom and volunteer study indicate that this method can perform registration in 34 ms with root-mean-square error of 1.6 mm when the given misalignment of a liver is 30 mm. The clinical studies demonstrate that the method can track liver motion of up to approximately 40 mm. Matching profiles with cross-correlation information perform better than with mutual information in terms of robustness and speed. CONCLUSION: The proposed image registration method has potential clinical impact on and advantages for MR-guided liver therapy. PMID- 14746412 TI - Alzheimer's disease: to tell or not to tell. PMID- 14746413 TI - Medical comorbidity in black and white patients with Alzheimer's disease. AB - BACKGROUND: Little is known about co-medical illnesses in black and white patients with probable Alzheimer's disease (AD). METHODS: To address this question, we used two methods. In the first (Group I), black and white probable AD patients were matched on age at presentation to the clinic, age of onset of AD, duration of illness, and Mini-Mental State Examination scores; then, a variety of co-medical illnesses were compared between blacks and whites. In Group II, whites were randomly matched to blacks on the variables listed above. RESULTS: In Group I, blacks were found to have a higher rate of hypertension than whites, whereas whites had a higher incidence of atrial fibrillation and cancer than blacks. In Group II, age at presentation to the clinic was found to be shorter for men than for women; duration of illness was shorter for black men than for white men, white women, and black women; and Mini-Mental State Examination scores were lower in blacks than whites. As in Group I, blacks were found to have a higher rate of hypertension, whereas whites had higher rates of atrial fibrillation, cancer, coronary artery disease, high cholesterol, and gastrointestinal disease. CONCLUSION: In both groups, black patients with probable AD had a higher rate of hypertension than white patients with probable AD, and whites had higher rates of atrial fibrillation and cancer. This finding suggests that these comorbid illnesses in black and white patients with probable AD is not due to a statistical Type II error, but rather to differences in these groups. PMID- 14746414 TI - Surgical treatment of achalasia in the 21st century. AB - BACKGROUND: Achalasia is a primary motility disorder of the esophagus characterized by poor mid-esophageal motility and failure of the lower esophageal sphincter to properly relax. The optimal treatment of the disease would improve esophageal peristalsis and promote lower esophageal sphincter relaxation. Currently, such therapy is not possible, so treatment of the disorder is aimed at relief of symptoms by disruption of the lower esophageal sphincter. METHODS: Data were collected prospectively on all patients undergoing laparoscopic myotomy and Toupet fundoplication during a 6-year period. RESULTS: Fifty-nine patients with a mean age of 44 years were treated during a 6-year period. Fifty-three patients underwent laparoscopic myotomy with Toupet fundoplication (91%), and four had laparoscopic myotomy without a fundoplication (6%). Fundoplication was not performed in two patients who had a megaesophagus. Two patients required conversion to an open operation. Sixty percent of patients were discharged the day after surgery; the average length of stay for all patients was 2.1 days. Ten percent of patients had minor complications; none required reoperation. Mortality was 0%, and 96% of patients rated their postoperative swallowing ability as excellent or good. CONCLUSION: Surgical myotomy is becoming first-line therapy for all patients with achalasia. A strong working relationship between surgeon and gastroenterologist helps to optimize patient care. PMID- 14746415 TI - Tonsillectomy using the Colorado microdissection needle: a prospective series and comparative technique review. AB - Tonsillectomy remains one of the most common surgical procedures performed worldwide. Recent advancements in equipment technology have ushered in several new tonsillectomy techniques. Among these is the Colorado tip electromicrodissection needle. In this report, we describe the technical aspects of this modality and report our results in a prospective study of 12 adults and 13 children. We then compare our results with several published series using a variety of techniques. We found tonsillectomy using the electromicrodissection needle compares most favorably in all criteria examined, including operative and perioperative blood loss, perioperative pain, return to regular diet, and cost. We conclude that electromicrodissection tonsillectomy is an excellent option for all surgeons performing tonsillectomy. PMID- 14746416 TI - Four-cut sinus computed tomographic scanning in screening for sinus disease. AB - OBJECTIVE: We sought to calculate the sensitivity, specificity, negative predictive value, and positive predictive value of a limited sinus computed tomographic (CT) scan for sinus disease. METHODS: We conducted a retrospective case series. Inclusion criteria included a sinus CT scan obtained between April 1999 and November 2000. From the complete sinus CT scan, the limited series were obtained by blocking from view all the other cuts and leaving the radiologist only four slices to read (midfrontal, anterior maxillary sinuses, posterior maxillary sinuses, and midsphenoidal). The complete CT scan was the "gold standard." RESULTS: Fifty-one patients were eligible. We observed 81.3% sensitivity, 89.5% specificity, a 73.9% negative predictive value, and a 92.9% positive predictive value for the limited CT scan for the detection of sinus disease. This sensitivity and specificity were higher than reported in the literature for plain films. CONCLUSION: The limited sinus CT scan is superior to plain radiographs but is not as good as the full CT scan in the evaluation of sinusitis. PMID- 14746418 TI - Handgrip strength, pulmonary function tests, and pulmonary muscle strength in fibromyalgia syndrome: is there any relationship? AB - OBJECTIVE: It has been reported that patients with fibromyalgia syndrome (FMS) have lower maximal respiratory pressures than healthy subjects, indicating reduced pulmonary muscle strength. It has also been reported that patients with FMS have reduced grip strength. In this study, we aimed to examine the possible relationship between handgrip strength as a determinant of peripheral muscle strength and pulmonary muscle strength in patients with FMS by comparing them with healthy controls. METHODS: Forty-one consecutive women with FMS (diagnosed according to the American College of Rheumatology 1990 criteria) were compared with 40 age- and body mass index-matched healthy female controls. Pulmonary function tests were assessed by spirometry. Maximal pulmonary pressures were evaluated using an oral pressure meter. A dyspnea score was obtained. Pain was scored according to visual analogue scale and chest pain was classified (0-9) in fibromyalgia patients. Chest expansion was also measured in the two groups. Tender points were also evaluated in FMS patients. Grip strength (Jamar handheld dynamometer) was also measured in the two groups. RESULTS: The difference in pulmonary function tests was not statistically significant between groups. Maximal respiratory pressures (maximum inspiratory pressure and maximum expiratory pressure) and endurance (maximum ventilatory volume) were significantly lower in patients with FMS than in controls. There was also a statistically significant difference between groups regarding grip strength. There was also significant correlation between maximal inspiratory pressure and maximal expiratory pressure values and handgrip strength in patients with FMS. CONCLUSION: These data indicate that handgrip strength may be a determinant of pulmonary muscle strength in fibromyalgia patients. PMID- 14746417 TI - Changes in transcutaneous carbon dioxide, oxygen saturation, and respiratory rate after interscalene block. AB - BACKGROUND: We used transcutaneous (TC) carbon dioxide (CO2) monitoring to prospectively evaluate changes in respiratory status after interscalene anesthesia in 45 adults (40 successful and 5 unsuccessful blocks). METHODS: TC CO2 oxygen saturation and respiratory rate were recorded every minute for 5 minutes before block and every 2 minutes for a total of 30 minutes (15 data sets) after injection of the local anesthetic solution. RESULTS: After successful block, TC-CO2 increased from 41 +/- 5 mm Hg to a maximum value of 44 +/- 6 mm Hg (P < 0.0001) and the respiratory rate increased from 14 +/- 2 breaths/min to a maximum of 20 +/- 4 breaths/min (P < 0.001). The increase in TC-CO2 was > or = 5 mm Hg in 11 patients and > or = 10 mm Hg in 4 patients, with a maximum increase of 12 mm Hg. Of the 600 TC-CO2 data points recorded (15 each from the 40 patients with a successful block), 62 (10.3%) showed a TC-CO2 value of 50 mm Hg or greater, with a maximum value of 57 mm Hg. No significant change in TC-CO2 or respiratory rate was seen in the five patients with unsuccessful block. CONCLUSION: After interscalene blockade, we found an increase in respiratory rate and hypercarbia that resulted in no clinically significant effect. PMID- 14746420 TI - Sex differences in analgesia: a randomized trial of mu versus kappa opioid agonists. AB - OBJECTIVES: We sought to evaluate whether there is a sex difference in the analgesic response to mu versus kappa opioids in the management of acute moderate to severe pain of injury in the emergency department. METHODS: The study was a randomized, double-blind, clinical trial comparing the prototypical mu-receptor agonist, morphine sulfate, to the prototypical kappa agonist, butorphanol. The primary endpoints were degree of relief by visual analog scores at 30 and 60 minutes. Statistical analysis was performed using Mann-Whitney Utest for nonparametric analysis and repeated-measures analysis of variance. RESULTS: Ninety-four patients were entered in the study, with 49 (52%) males and 45 (48%) females. Forty-six received morphine sulfate and 48 received butorphanol. There was no difference in demographics in the two groups. At 60 minutes, females had significantly lower visual analog scores with butorphanol compared with morphine (P = 0.046). At 60 minutes, there was a trend for a difference in response of males versus females to morphine, with males responding better than females (P = 0.06). CONCLUSION: Females had better pain scores with butorphanol than morphine at 60 minutes. PMID- 14746419 TI - Prevalence of occult celiac disease in patients with iron-deficiency anemia: a prospective study. AB - BACKGROUND: Occult celiac disease has been reported in 0 to 6% of adults presenting with iron-deficiency anemia. Most prior studies have been retrospective or screened only a selected population of patients with small bowel biopsies. To more accurately define the true prevalence of this disorder in patients presenting with iron-deficiency anemia (with or without stool hemoccult positivity), we initiated this prospective study. METHODS: Esophagogastroduodenoscopy with small bowel biopsies and colonoscopy were performed in all iron-deficiency anemia patients (including those with hemoccult positive stools) referred to the gastroenterology service during a 2-year period (1998-2000). Inclusion criteria included iron-deficiency anemia as defined by a serum ferritin < 25 ng/ml and anemia with hemoglobin < 12 g/dl. Patients were excluded for documented prior erosive, ulcerative, or malignant disease of the gastrointestinal tract, previous gastrointestinal surgery, overt gastrointestinal bleeding within the past 3 months, or inability to access the duodenum for biopsy. All patients underwent upper endoscopy with more than two biopsies of the distal duodenum and colonoscopy. A serum immunoglobulin A antiendomysial antibody test was to be performed in those patients with a positive small bowel biopsy to confirm the diagnosis of celiac disease. RESULTS: One hundred five of 139 consecutive patients with iron-deficiency anemia met the inclusion criteria and were enrolled in the study. Fifty-seven men (mean age, 51.6 yr) and 48 women (mean age, 54.1 yr) constituted the study population. The demographics of this study population included 36 blacks, 38 Hispanics, and 22 whites. Nine patients were of mixed or unknown ethnic background. Forty-three and eight-tenths percent of the men and 37.5% of women had hemoccult-positive stools, accounting for a total of 40.9% of the study patients. Upper endoscopic findings included gastritis in 22.8%, gastric ulcers in 9.5%, duodenitis in 8.5%, esophagitis in 7.6%, Barrett's ulcer in 2.8%, duodenal ulcer in 2.8%, gastric polyp in 2.8%, and celiac disease in 2.8%. Colonoscopic findings included colon polyps in 21.9%, diverticula in 10.4%, and hemorrhoids in 16.1%. Multiple findings were found in 32.3% of patients, and there were no findings in 28.5% of patients. CONCLUSION: The prevalence of occult celiac disease in this prospective study of patients presenting with iron-deficiency anemia was 2.8%. A significant number of other gastrointestinal lesions amenable to therapy were also found on upper and lower endoscopy in these patients. Given the treatable nature of celiac disease, it should be screened for in patients with unexplained iron-deficiency anemia with or without hemoccult-positive stools. PMID- 14746421 TI - Intralesional interferon-alpha-2B injections for the treatment of Peyronie's disease. AB - BACKGROUND: Intralesional therapies provide an alternative to surgical treatment of Peyronie's disease (PD). This study examines the efficacy of intraplaque injections of interferon-alpha-2B (IFN-alpha-2B) in the treatment of PD. METHODS: Twenty-five patients were enrolled in the study with 21 completing the study. The average age of the study participants was 55.8 years, with 10 of the 21 having diabetes, hypertension, or both. Seven patients received placebo injections using 10 ml saline biweekly for 6 weeks before interferon therapy, and all 21 patients received biweekly injections of 2 x 10(6) U IFN-alpha-2B for 6 weeks before reevaluation. Patients were evaluated with duplex Doppler ultrasonography to assess penile curvature and blood flow. RESULTS: The International Index of Erectile Function questionnaire was completed by 14 of 21 men to assess severity of erectile dysfunction (ED) before and after treatment. Improvements of 20% or more in penile curvature occurred in 14 (67%) of 21 men. Penile pain decreased in eight (80%) of 10, and plaque size decreased subjectively in 15 (71%) of 21 participants. The International Index of Erectile Function questionnaire showed significant improvement of ED in five (71%) of seven men with baseline moderate to severe ED. Peak systolic blood flow improved in three (43%) of seven men with baseline hemodynamic impairment, and veno-occlusive disease resolved in three (18%) of 17 individuals. IFN-alpha-2B injections led to significant improvements in penile pain and curvature caused by PD. CONCLUSION: Results of intralesional IFN-alpha-2B injections in improving ED are encouraging. The study findings warrant further investigation of the effectiveness of IFN-alpha-2B injections for treatment of PD. PMID- 14746422 TI - Cardiopathia fantastica: the cardiac variant of Munchausen syndrome. AB - Munchausen syndrome is a rare condition in which the patient repeatedly seeks medical care for factitious illnesses. Cardiac Munchausen syndrome was first reported in 1953 and later referred to as cardiopathia fantastica. It is characterized by clinical manifestations of acute cardiac disease that are feigned and recurrent. Cardiopathia fantastica can have a variety of presentations similar to true cardiac disease. Most cases have presented with chest pain simulating acute coronary artery disease, but arrhythmias, hypertensive crises, abnormal biochemistry, and electrocardiographic findings have also been noted. These patients are willing to undergo expensive, invasive, and risky procedures to evaluate their simulated illness. This condition is likely to be significantly underreported. In some patients, the presence of abnormal findings that are clinically insignificant may complicate the investigative approach. Patients with this disorder consume a disproportionate amount of health care dollars and sometimes are left with residual deficits as complications of invasive procedures. In this review, we discuss the recognition, manifestations, and treatment of cardiopathia fantastica. PMID- 14746423 TI - Vasectomy: a "seminal" analysis. AB - Vasectomy is one of the most reliable and cost-effective permanent methods of contraception. Despite its popularity, certain issues pertaining to the procedure remain unresolved. Appropriate selection of candidates for vasectomy requires thorough counseling and screening, though a foolproof method of eliminating dissatisfaction and regret remains a mystery. Debate continues over the relative merits of the various techniques of isolating and sealing the vasal ends. Postoperative complication rates remain minimal regardless of the technique used, and no single strategy attempting to maximize patient compliance with postoperative semen analysis has enjoyed unmitigated success. Long-term consequences, other than regret, are rare. Finally, issues regarding residency training in the procedure and its impact on procedure morbidity are scarcely addressed in the literature. This study reviews the evolution of vasectomy as a contraceptive procedure and attempts to summarize current literature addressing these unresolved issues. PMID- 14746424 TI - Cerebellar involvement in legionellosis. AB - The involvement of the cerebellum in Legionnaires' disease has been noted in occasional case reports, but there have been no reviews on this subject to date. We report a previously healthy patient who contracted Legionella pneumonia and developed profound cerebellar dysfunction during his illness. He was treated with antibiotics with improvement of his pneumonia, but his cerebellar symptoms persisted. We review 29 case reports of cerebellar dysfunction in Legionnaires' disease and summarize the clinical course, cultures, cerebrospinal fluid analyses, and neuroimaging. Finally, possible methods of pathogenicity are discussed including data regarding direct bacterial invasion, toxin production, and immune-mediated mechanisms. PMID- 14746425 TI - Capgras syndrome related to diazepam treatment. AB - Capgras syndrome, the delusion that identical-appearing impostors have replaced familiar people, is an unusual phenomenon usually seen in schizophrenia or dementia. We recently cared for a 78 year old man who seemed to develop Capgras syndrome as an adverse reaction to diazepam. An iatrogenic cause should be considered in the differential diagnosis of any new delusion, including Capgras syndrome. PMID- 14746426 TI - Celiac disease as a manifestation of Munchausen by proxy. AB - In typical cases of Munchausen by proxy maltreatment, a mother feigns or produces illness in her child. Her primary goal is to accrue emotional gratification, and no mental disorder better accounts for the behavior. We present the first published case in which the principal manufactured ailment was celiac sprue. In addition, a panoply of other ailments ranging from seizures to behavioral abnormalities was reported. The case is also very unusual in the involvement of the paternal grandmother and, to a lesser extent, the paternal grandfather as the perpetrators. Although definitive intervention to protect the child occurred only after 7 years had passed, multidisciplinary teamwork ultimately resulted in a successful outcome for the child, who is now doing well. PMID- 14746427 TI - Medication-associated depersonalization symptoms: report of transient depersonalization symptoms induced by minocycline. AB - Patients with depersonalization disorder experience episodes in which they have a feeling of detachment from themselves. Symptoms of depersonalization may occur in individuals who have other mental disorders, or who have various medical conditions, or who have taken certain medications. A woman developed depersonalization symptoms after initiation of minocycline therapy. Her symptoms ceased after treatment was stopped and recurred when she restarted the drug. Medications that have been associated with causing symptoms of depersonalization are presented and the postulated pathogenesis by which some of these drugs induced depersonalization symptoms is discussed. Medication-associated depersonalization symptoms typically resolve once the inducing drug has been withdrawn. PMID- 14746428 TI - Septic arthritis caused by Chryseobacterium meningosepticum in an elbow joint prosthesis. AB - Chryseobacterium meningosepticum is a Gram-negative bacillus historically associated with meningitis and sepsis in premature neonates. Clinicians should suspect this organism when Gram-negative bacilli are seen on Gram-stain and culture, particularly in immunocompromised patients, and in cases of disrupted host tissue integrity. We report the first case of septic arthritis due to this organism. PMID- 14746429 TI - Septic arthritis of the ankle due to Salmonella enteritidis: a case report. AB - Salmonella septic arthritis in healthy individuals is a rare phenomenon in the United States. This case report chronicles the clinical course of a 41-year-old male farmworker who presented with a 3-week history of fever, chills, night sweats with pain, and swelling and redness of his left ankle. He had an open fracture of the ankle 2 years earlier that healed and was asymptomatic despite prior radiographic evidence of avascular necrosis of the talar dome. One month before presentation, he had an ipsilateral periungual abscess of the great toe that he opened and drained himself. Joint cultures were positive for Salmonella enteritidis that was successfully treated with a 6-week course of i.v. ceftriaxone. PMID- 14746430 TI - Pigmented villonodular synovitis of the knee in a 9-year-old child. AB - This report describes a 9-year-old girl with pigmented villonodular synovitis of the right knee. She presented with a chronic bloody effusion of the knee. The diagnosis of pigmented villonodular synovitis was suggested by the findings on magnetic resonance imaging and confirmed at the time of arthroscopic synovectomy. This report emphasizes the importance of considering pigmented villonodular synovitis in the differential diagnosis of chronic hemarthrosis in children. PMID- 14746431 TI - Neck pain: common complaint, uncommon diagnosis--symptomatic clival chordoma. AB - Patients presenting with neck complaints, such as pain or stiffness, are not uncommon in the Emergency Department. Complaints of neck instability, however, are unusual. We report the case of a 30-year-old woman who presented with multiple neck complaints that included having a "wobbly" sensation of her neck on flexion, feeling as if it were unstable. Our patient indeed had atlanto-occipital instability secondary to a locally destructive tumor of the cranial base, known as a clival chordoma. Chordomas are rare and unique bony tumors that arise along the neural axis and are thought to originate from the nucleus pulposus. The tumors are slow growing; locally invasive; and cause a variety of neurologic, musculoskeletal, cranial, and neck complaints. We describe this unique case and its presentations in an attempt to increase the sensitivity of physicians in early detection of this rare and lethal tumor. PMID- 14746432 TI - Risk-factor profile in severe, generalized, obliterating vascular disease. AB - A 74-year-old woman had a history over 25 years of endarterectomy of both renal arteries, iliac venous thrombosis, pulmonary embolism, left internal carotid artery endarterectomy, coronary angioplasty, aortocoronary bypass grafting, occlusion of the right axillary artery, lower-limb claudication due to common iliac artery aneurysm, external iliac artery stenosis, multiple femoral artery stenoses, bifurcational stent grafting, occlusion of the left brachial artery and the right external iliac artery, and stroke. Assessment of the risk-factor profile revealed an absence of classic risk factors but the presence of the factor V Leiden mutation, the methylenetetrahydrofolate reductase AI298C mutation, the HFE C282Y mutation, plasminogen activator inhibitor-1 gene mutation, the -455 G/A fibrinogen gene polymorphism, the epsilon3/epsilon4 apolipoprotein E -675 4G gene polymorphism, and hyperhomocysteinemia. This case shows that severe, generalized, occlusive vascular disease may be due to the combination of various genetic risk factors for atherosclerosis and venous thromboembolism. PMID- 14746433 TI - Esophageal foreign body aspiration presenting as asthma in the pediatric patient. AB - This report describes three cases of foreign body ingestion incorrectly diagnosed as asthma and/or upper respiratory tract infection. These cases demonstrate the need for further clinical investigations in any child who does not improve with adequate therapy. Many of our patients had symptoms of asthma and/or respiratory tract infection for at least 1 month before correct diagnosis. PMID- 14746434 TI - Cytomegalovirus enteritis in common variable immunodeficiency. AB - A woman with previously undiagnosed common variable immunodeficiency presented with diarrhea and volume depletion. Biopsies from upper and lower endoscopy revealed atrophic gastritis, villous atrophy, and an inflammatory bowel disease like chronic colitis, with absence of plasma cells in all sites. Cytomegalovirus inclusions were demonstrated in the colon and small bowel mucosa. Despite therapy with intravenous immunoglobulin and ganciclovir, the patient deteriorated rapidly and subsequently died. This case report highlights the potential for cytomegalovirus to cause extensive disease in patients with common variable immunodeficiency and, thus, the importance of considering it in the initial differential diagnosis so that further morbidity and mortality might be prevented. PMID- 14746435 TI - Hyalinizing Spitz nevus: spindle and epithelioid cell nevus with paucicellular collagenous stroma. AB - Hyalinizing spindle and epithelioid cell nevus (Spitz nevus) is an uncommon melanocytic lesion. The histologic features of this benign tumor can mimic those of certain benign (dermatofibroma and desmoplastic cellular blue nevus) and malignant (metastatic carcinoma and malignant melanoma) neoplasms. We report a woman with a hyalinizing Spitz nevus and review the clinical characteristics, histologic features, and differential diagnosis of this lesion. The lesion presented as an asymptomatic red firm nodule on her abdomen. Microscopic examination showed a symmetric lesion in the dermis. The neoplastic cells were large and epithelioid-shaped or plump and spindle-shaped. They were present as isolated individual cells, single cells in a linear pattern, small nests, and fascicles in a paucicellular hyalinized stroma. Neoplastic cells with prominent eosinophilic nucleoli were evenly exhibited from the superficial to deep lesion. Diffuse expression of S-100 protein and absence of staining with antibodies to cytokeratin and HMB-45 were observed. The features were diagnostic of a hyalinizing Spitz nevus. Hyalinizing Spitz nevus is a benign lesion whose histologic features may mimic dermatofibroma, desmoplastic cellular blue nevus, metastatic carcinoma, and malignant melanoma. Immunohistochemical studies are helpful to differentiate these lesions. PMID- 14746436 TI - Official positions of the International Society for Clinical Densitometry. PMID- 14746437 TI - Analytic strategies for longitudinal studies. PMID- 14746438 TI - Deviations from the population-averaged versus cluster-specific relationship for clustered binary data. AB - There has been much debate about the relative merits of mixed effects and population-averaged logistic models. We present a different perspective on this issue by noting that the investigation of the relationship between these models for a given dataset offers a type of sensitivity analysis that may reveal problems with assumptions of the mixed effects and/or population-averaged models for clustered binary response data in general and longitudinal binary outcomes in particular. We present several datasets in which the following violations of assumptions are associated with departures from the expected theoretical relationship between these two models: 1) negative intra-cluster correlations; 2) confounding of the response-covariate relationship by cluster effects; and 3) confounding of autoregressive relationships by the link between baseline outcomes and subject effects. Under each of these conditions, the expected theoretical attenuation of the population-averaged odds ratio relative to the cluster specific odds ratio does not necessarily occur. In all cases, the naive fitting of a random intercept logistic model appears to lead to bias. In response, the random intercept model is modified to accommodate negative intra-cluster correlations, confounding due to clusters, or baseline correlations with random effects. Comparisons are made with GEE estimation of population-averaged models and conditional likelihood estimation of cluster-specific models. Several examples, including a cross-over trial, a multicentre nonrandomized treatment study, and a longitudinal observational study are used to illustrate these modifications. PMID- 14746439 TI - Instrumental variables and inverse probability weighting for causal inference from longitudinal observational studies. AB - Inferring causal effects from longitudinal repeated measures data has high relevance to a number of areas of research, including economics, social sciences and epidemiology. In observational studies in particular, the treatment receipt mechanism is typically not under the control of the investigator; it can depend on various factors, including the outcome of interest. This results in differential selection into treatment levels, and can lead to selection bias when standard routines such as least squares regression are used to estimate causal effects. Interestingly, both the characterization of and methodology for handling selection bias can differ substantially by disciplinary tradition. In social sciences and economics, instrumental variables (IV) is the standard method for estimating linear and nonlinear models in which the error term may be correlated with an observed covariate. When such correlation is not ruled out, the covariate is called endogenous and least squares estimates of the covariate effect are typically biased. The availability of an instrumental variable can be used to reduce or eliminate the bias. In public health and clinical medicine (e.g., epidemiology and biostatistics), selection bias is typically viewed in terms of confounders, and the prevailing methods are geared toward making proper adjustments via explicit use of observed confounders (e.g., stratification, standardization). A class of methods known as inverse probability weighting (IPW) estimators, which relies on modeling selection in terms of confounders, is gaining in popularity for making such adjustments. Our objective is to review and compare IPW and IV for estimating causal treatment effects from longitudinal data, where the treatment may vary with time. We accomplish this by defining the causal estimands in terms of a linear stochastic model of potential outcomes (counterfactuals). Our comparison includes a review of terminology typically used in discussions of causal inference (e.g., confounding, endogeneity); a review of assumptions required to identify causal effects and their implications for estimation and interpretation; description of estimation via inverse weighting and instrumental variables; and a comparative analysis of data from a longitudinal cohort study of HIV-infected women. In our discussion of assumptions and estimation routines, we try to emphasize sufficient conditions needed to implement relatively standard analyses that can essentially be formulated as regression models. In that sense this review is geared toward the quantitative practitioner. The objective of the data analysis is to estimate the causal (therapeutic) effect of receiving combination antiviral therapy on longitudinal CD4 cell counts, where receipt of therapy varies with time and depends on CD4 count and other covariates. Assumptions are reviewed in context, and resulting inferences are compared. The analysis illustrates the importance of considering the existence of unmeasured confounding and of checking for 'weak instruments.' It also suggests that IV methodology may have a role in longitudinal cohort studies where potential instrumental variables are available. PMID- 14746440 TI - Functional data analysis in longitudinal settings using smoothing splines. AB - Data in many experiments arise as curves and therefore it is natural to use a curve as a basic unit in the analysis, which is termed functional data analysis (FDA). In longitudinal studies, recent developments in FDA have extended classical linear models and linear mixed effects models to functional linear models (also termed varying-coefficient models) and functional mixed effects models. In this paper we focus our review on the functional mixed effects models using smoothing splines, because functional linear models are special cases of this more general framework. Due to the connection between smoothing splines and linear mixed effects models, functional mixed effects models can be fitted using existing software such as SAS Proc Mixed. A case study is presented as an illustration. PMID- 14746441 TI - Mixed effects multivariate adaptive splines model for the analysis of longitudinal and growth curve data. AB - In this article, I review the use of nonparametric methods in the analysis of longitudinal and growth curve data, particularly the multivariate adaptive splines models for the analysis of longitudinal data (MASAL). These methods combine nonparametric techniques (B-splines, kernel smoothing, piecewise polynomials) and models with random effects, and provide fruitful alternatives to mixed effects linear models. Similarities, differences, strengths and limitations among these methods are presented. The analysis of a real example is also presented to illustrate the application and interpretation of MASAL. Open questions are posed for further investigation. PMID- 14746442 TI - High mobility of dithiophene-tetrathiafulvalene single-crystal organic field effect transistors. AB - Single-crystal field effect transistors of the organic semiconductor dithiophene tetrathiafulvalene (DT-TTF) were prepared by drop casting. Long, thin crystals connected two microfabricated gold electrodes, and a silicon substrate was used as a back gate. The highest hole mobility observed was 1.4 cm2/Vs, which is the highest reported for an organic semiconductor not based on pentacene. A high ON/OFF ratio of at least 7 x 105 was obtained for this device. PMID- 14746443 TI - A new class of "tethered" ruthenium(II) catalyst for asymmetric transfer hydrogenation reactions. AB - Ruthenium dimer 4 is converted directly to monomeric asymmetric transfer hydrogenation catalyst 2 under the conditions employed for ketone reduction. Using 0.25 mol % of either 4 or 0.5 mol % of 2 in formic acid/triethylamine, it is possible to achieve ketone reduction in quantitative conversion and with ee's as high as 98%. Complex 2 is a robust "single-reagent" catalyst which offers significant scope for modification toward specific substrates. The synthesis and applications of an analogous complex derived from (1R,2S)-norephedrine are also described. PMID- 14746444 TI - A condensable amphiphile with a cleavable tail as a "Lizard" template for the sol gel synthesis of functionalized mesoporous silica. AB - Use of a dialkoxysilane-containing, condensable alanine amphiphile with a cleavable hexadecyl ester tail (1) as a "lizard" template for sol-gel synthesis results in the formation of a mesoporous silicate (2) whose channels are filled with an organic group of the template. Treatment of 2 with aqueous HCl allows selective cleavage of the ester moiety to leave nanochannels (3) whose surface is densely covered with alanine-CO2H. According to XRD and TEM, 2, on conversion into 3, can preserve its regular hexagonal structure. 3 displays a clear N2 adsorption/desorption isotherm with a BET surface area of 536 m2 g-1. 3 can also adsorb a basic guest such as NH3 up to 0.7 mmol g-1, which is 7 times greater than that observed for mesoporous silica obtained by calcination of 2. PMID- 14746445 TI - Labeling vibrations by light: ultrafast transient 2D-IR spectroscopy tracks vibrational modes during photoinduced charge transfer. AB - We report on a novel ultrafast two-dimensional infrared laser experiment that correlates vibrational bands of reactant and product of a photoreaction. The possibilities of this technique are demonstrated for the metal-to-ligand charge transfer (MLCT) in [Re(CO)3Cl(dmbpy)] (dmbpy = 4,4'-dimethyl-2,2'bipyridine) where we correlated the CO vibrational modes of the ground state and the MLCT state. A distinct vibrational mode is excited in the electronic ground state by an infrared laser pulse. This vibrational label survives the subsequent electronic excitation and can be followed in the excited electronic state. It is shown that the order of the vibrational energy levels is not preserved when exciting the molecule as was commonly assumed in the literature. PMID- 14746446 TI - Desulfurization of diesel fuels by adsorption via pi-complexation with vapor phase exchanged Cu(I)-Y zeolites. AB - Desulfurization of a commercial diesel fuel by vapor-phase ion exchange (VPIE) copper(I) faujasite zeolites was studied in a fixed-bed adsorber operated at ambient temperature and pressure. The zeolite adsorbed approximately five thiophenic molecules per unit cell. After treating 18 cm3 of fuel, the cumulative average sulfur concentration detected was 0.032 ppmw-S. GC-FPD results showed that the pi-complexation sorbents selectively adsorbed highly substituted thiophenes, benzothiophenes, and dibenzothiophenes from diesel, which is not possible by using conventional hydrodesulfurization (HDS) reactors. The high sulfur selectivity and high sulfur capacity of the VPIE Cu(II)-zeolites were due to pi-complexation. PMID- 14746447 TI - One-dimensional ion transport in self-organized columnar ionic liquids. AB - New fan-shaped ionic liquids forming columnar liquid crystalline phases have been prepared to obtain one-dimensional ion-transporting materials. The ionic liquids consist of two incompatible parts: an imidazolium-based ionic part as an ion conducting part and tris(alkyloxy)phenyl parts as insulating parts. Two compounds having octyl and dodecyl chains have been synthesized. Self-assembly of these materials leads to the formation of thermotropic hexagonal columnar liquid crystalline states at room temperature. Anisotropic one-dimensional ionic conductivities have been successfully measured by the cells having comb-shaped gold electrodes. The self-organized columns have been aligned macroscopically in two directions by shearing perpendicular and parallel to the electrodes. The ionic conductivities parallel to the column axis are higher than those perpendicular to the axis. The incorporation of lithium salts in these columnar materials leads to the enhancement of the ionic conductivities and their anisotropy. These materials would be useful for anisotropic transportation of ions at the nanometer level. PMID- 14746448 TI - From evolution to green chemistry: rationalization of biomimetic oxygen-transfer cascades. AB - Thermodynamic electron-transfer potentials from biology textbooks elucidate the sequence of electron-transfer events in the respiratory chain in mitochondria. In this study, thermodynamic and kinetic oxygen-transfer potentials have been defined and predicted for oxidants and substrates using density functional theory, aiming to rationalize multiple oxygen-transfer events in chemical catalysis, particularly in current developments of the Sharpless dihydroxylation. Key transition states for competing mechanisms in a recent dihydroxylation method containing the olefin, osmium tetraoxide, methyltrioxorhenium(VII), a chiral tertiary amine, and the green terminal oxidant hydrogen peroxide have been investigated rigorously. The calculations show the amine to function as an oxygen transfer mediator between rhenium peroxides and osma-2,5-dioxolanes, in addition to its role as a carrier of chiral information. Unique mechanistic and stereoelectronic patterns in this oxygen-transfer cascade explain the unexpected failure of reactivity predictions using simpler models such as Marcus theory. PMID- 14746449 TI - Synthesis of amphidinolide T1 via catalytic, stereoselective macrocyclization. AB - Two nickel-catalyzed reductive coupling reactions of alkynes were instrumental in a modular, stereoselective synthesis of amphidinolide T1 (1). The C13-C21 fragment was prepared from two simple starting materials that were joined in a catalytic alkyne-epoxide fragment coupling operation, whereas an intramolecular aldehyde-alkyne reductive coupling simultaneously formed the final carbon-carbon bond of the macrocycle and established the C13 carbinol configuration with complete selectivity in the desired fashion. PMID- 14746450 TI - Generalized reconstruction of n-D NMR spectra from multiple projections: application to the 5-D HACACONH spectrum of protein G B1 domain. AB - Reconstructing multidimensional NMR spectra from 2-D projections significantly reduces the time needed for data collection over conventional methodology. Here, we provide a generalization of the projection-reconstruction process to spectra of arbitrary dimensionality, using a concept of coordinate rotation to produce explicit expressions for reconstruction. These expressions allow one to reconstruct subsets of the higher dimensionality space without producing the full spectrum, permitting convenient analysis of the data. We demonstrate the effectiveness of these procedures in the reconstruction of the 5-D HACACONH spectrum of protein G B1 domain from 12 2-D projections collected in five experiments. We further demonstrate that the base spectra of GFT-NMR are equivalent to projections of the 5-D spectrum at fixed angles. PMID- 14746451 TI - Glycosidic bond cleavage of thymidine by low-energy electrons. AB - Thymidine was exposed to low-energy electrons (LEE) as a thin solid film under a high vacuum. Nonvolatile radiation products, remaining on the irradiated surface, were analyzed by HPLC/UV and GC/MS. Here, we show that exposure of thymidine to 3 100 eV electrons gives thymine as a major product with a yield of 3.2 x 10-2 per electron (about one-third of the total decomposition of thymidine). The formation of thymine indicates that LEE induces cleavage of the glycosidic bond separating the base and sugar moieties, suggesting a nonionizing resonant process involving dissociative attachment (<15 eV). In contrast, this reaction is not very efficient by DNA base ionization and does not occur by the reaction of solvated electrons with DNA. These studies introduce a new mechanism of DNA damage involving the interaction of LEE. PMID- 14746452 TI - Preparation of micro- and nanopatterns of polymer chains grafted onto flexible polymer substrates. AB - In this work a simple novel method for preparing micro- and nanoscale patterns of polymer chains grafted onto flexible polymer substrates is described. A combination of the two techniques of radiation grafting and "grafting-from" has been made. This combination makes it possible to prepare grafted structures having micro- or nanoscale lateral dimensions that are determined by the electron beam or X-ray irradiation patterns used. The height of the grafted features can be controlled by the irradiation dose or such grafting reaction conditions as time, temperature, or monomer concentration. Our first results for nanopatterned samples demonstrate resolution comparable to those of other polymer-based lithography processes. PMID- 14746453 TI - Thermal hysteresis in the photoresponsivity of a langmuir film of amphiphilic spiropyran. AB - We have found a thermal hysteresis in the photoresponsivity of a Langmuir film for the first time. The Langmuir film of an amphiphilic spiropyran, 1',3'-dihydro 3',3'-dimethyl-6-nitro-1'-octadecyl-8-(docosanoyloxymethyl)spiro[2H-1-benzopyran 2,2'-(2H)-indole] (SP) was fabricated at 13 degrees C at 10 mN m-1, followed by heating to a given temperature. UV irradiation of this film caused only the isomerization of SP to the corresponding merocyanine (MC) up to 29 degrees C. Light-induced J-aggregation of MC occurred at 30 degrees C. On the other hand, once the film was heated to 30 degrees C, light-induced J-aggregation was observed down to 27 degrees C. The hysteresis should be related with the phase transitions that occur in the bulk of SP at similar temperatures. No significant morphological change occurred by light-induced J-aggregation in the Langmuir Blodgett (LB) film of SP by the present method, in contrast to the case of the LB films fabricated under isothermal conditions at 30 degrees C. This feature enabled us to pattern the LB film with J-aggregate of MC by UV irradiation through a photomask of lines with a width of 5 mum each. PMID- 14746454 TI - Mutation of two active-site residues converts a phosphatidylinositol-specific phospholipase C to a glucose phosphatase. AB - Two mutations, R69D and K115E, converted a bacterial phosphatidylinositol specific phospholipase C (PI-PLC) to a phosphatase with much higher specific activity toward glucose-6-phosphate than inositol-1-phosphate. PI-PLC single mutations R69D and K115E can cleave PI but lack any demonstrable phosphatase activity. The bacterial PI-PLC has no sequence homology with known glucose-6 phosphatase enzymes, which need His, Arg, and negatively charged residues (Asp or Glu) at the active site. The change in chemical reaction and substrate specificity can be rationalized by energy minimization of the mutant with I-1-P or G-6-P bound. PMID- 14746455 TI - Controlling the selectivity of competitive nitroaldol condensation by using a bifunctionalized mesoporous silica nanosphere-based catalytic system. AB - A series of bifunctionalized mesoporous silica nanosphere-based (MSN) heterogeneous catalysts for the nitroaldol (Henry) reaction have been synthesized. A common 3-[2-(2-aminoethylamino)ethylamino]propyl (AEP) primary group and three different secondary groups, ureidopropyl (UDP), mercaptopropyl (MP), and allyl (AL) functionalities, were incorporated to these mesoporous silica materials by introducing equal amounts of AEP-trimethoxysilane with UDP-, MP-, or AL-trialkoxysilane precursors to our previously reported co-condensation reaction. Structures and relative concentrations of the functional groups were detailed by solid-state NMR and other spectroscopic techniques. The AEP group served as a catalyst, and the other secondary groups provided different noncovalent interactions to reactants and thereby controlled the reaction selectivity. By varying the secondary group in these bifunctionalized MSN catalysts, we investigated the selectivity of a nitroaldol reaction of two competing benzaldehydes reacting with nitromethane by measuring the molar ratio of the nitroalkene products. The selectivity of the bifunctionalized MSN catalysts could be systematically tuned simply by varying the physicochemical properties of the pore surface-bound secondary groups, i.e., polarity and hydrophobicity. PMID- 14746456 TI - Comparison of activation parameters for ionization reactions within zeolites and in aqueous solution. AB - Absolute rate constants for the ionization of chloride from the 2-chloro-1-(4 methoxyphenyl)ethyl radical are measured in aqueous methanol and in alkali-metal cation zeolites as a function of temperature. The absolute rate constants are very fast in the two distinct media. However, the activation parameters are considerably different. In solution, the reaction proceeds with low enthalpies of activation and large, negative entropies of activation, while in zeolites, the reaction is characterized by significantly higher activation enthalpies and large, positive entropies of activation. These differences reveal that the fundamental factors allowing for such rapid reactions are not the same in the two media. PMID- 14746457 TI - Selective interactions of porphyrins with semiconducting single-walled carbon nanotubes. AB - A derivatized porphyrin with long alkyl chains, 5,10,15,20 tetrakis(hexadecyloxyphenyl)-21H,23H-porphine, is selective toward semiconducting single-walled carbon nanotubes (SWNTs) in presumably noncovalent interactions, resulting in significantly enriched semiconducting SWNTs in the solubilized sample and predominantly metallic SWNTs in the residual solid sample according to Raman, near-IR absorption, and bulk conductivity characterizations. PMID- 14746458 TI - Mono- and diaminocarbenes from chloroiminium and -amidinium salts: synthesis of metal-free bis(dimethylamino)carbene. AB - Bis(trimethylsilyl)mercury cleanly reacts at low temperature with chloroamidinium and -iminium chlorides, generating persistent metal-free cyclic and acyclic diaminocarbenes, as well as transient aryl-, chloro-, and hydrogenoaminocarbenes; with the latter, the corresponding olefin dimers were isolated, whereas with the former, no dimerization processes were observed. PMID- 14746459 TI - C-H bond activation of hydrocarbons by an imidozirconocene complex. AB - Monomeric imidozirconocene complexes of the type Cp2(L)Zr=NCMe3 (Cp = cyclopentadienyl, L = Lewis base) have been shown to activate the carbon-hydrogen bonds of benzene, but not the C-H bonds of saturated hydrocarbons. To our knowledge, this singularly important class of C-H activation reactions has heretofore not been observed in imidometallocene systems. The M=NR bond formed on heating the racemic ethylenebis(tetrahydro)indenyl methyl tert-butyl amide complex, however, cleanly and quantitatively activates a wide range of n-alkane, alkene, and arene C-H bonds. Mechanistic experiments support the proposal of intramolecular elimination of methane followed by a concerted addition of the hydrocarbon C-H bond. Products formed by activation of sp2 C-H bonds are generally more thermodynamically stable than those formed by activation of sp3 C H bonds, and those resulting from reaction at primary C-H bonds are preferred over secondary sp3 C-H activation products. There is also evidence that thermodynamic selectivity among C-H bonds is sterically rather than electronically controlled. PMID- 14746460 TI - A quantum mechanics-based scoring function: study of zinc ion-mediated ligand binding. AB - In this communication, we report the development of a novel quantum mechanics based scoring function to predict free energy of ligand binding in the zinc metalloenzymes carbonic anhydrase (CA) and carboxypeptidase A (CPA). In particular, the AM1 method is used in conjunction with solvation modeling to predict the relative binding affinities of 18 CA and 5 CPA inhibitors. The effect of metal-ligand charge transfer is also discussed and shown to be different in CPA and CA, providing a further challenge to computing metalloenzyme binding affinities. PMID- 14746461 TI - Direct detection of oxygen intermediates in the non-heme Fe enzyme taurine/alpha ketoglutarate dioxygenase. AB - The reaction of substrate-bound taurine/alpha-ketoglutarate dioxygenase with O2 has been studied using cryogenic continuous-flow spectroscopy. Transient absorption spectra acquired at -38 degrees C show an exponential decay of a 318 nm chromophore with an apparent rate of 1.3 s-1. The observed optical changes and their kinetics are consistent with the profile of an Fe(IV) species detected recently by Mossbauer spectroscopy (Price et al., Biochemistry 2003, 42, 7497 7508). Resonance Raman measurement upon excitation at 363.7 nm reveal at least two oxygen isotope-sensitive vibrations at 821/787 cm-1 and 583/555 cm-1 for 16O and 18O derivatives, respectively. An additional mode is likely to be obscured by an ethylene glycol vibration at 865 cm-1 and/or 1089 cm-1. The 821 cm-1 vibration is assigned to the stretching mode of Fe(IV)=O species on the basis of its frequency and isotopic shift amplitude. The 583 cm-1 band is likely to originate from an Fe-O2 precursor of the Fe(IV)=O species, although its structural details are unclear at present. PMID- 14746463 TI - Stereocontrolled polymerization of racemic lactide with chiral initiator: combining stereoelection and chiral ligand-exchange mechanism. AB - New opportunities, provided by the 2,2'-[1,1'-binaphthyl-2,2' diylbis(nitrylomethilidyne)]diphenol (SB(OH)2)/Al(OiPr)3/racemic lactide (rac-LA) polymerization system, employing a combination of stereoelection with (S) and (R) ligand-exchange mechanism at Al-alkoxide active centers were explored. The stereoelectivity was comparable to that determined for the process with an additional synthetic step of isolation and purification of the SBO2Al-OiPr alkoxide. The resultant poly(rac-LA) had a gradient stereocopolymer structure and exhibited enhanced thermal stability due to a stereocomplex formation (Tm = 210 degrees C). This is the highest melting temperature reported until now for poly(lactide) (PLA) prepared directly from rac-LA. PMID- 14746462 TI - Direct synthesis of sulfonamides and activated sulfonate esters from sulfonic acids. AB - A general new method for the preparation of sulfonamides and activated sulfonate esters by the direct coupling of sulfonic acid salts with amines and alcohols using the reagent triphenylphosphine ditriflate is described. A new reusable polymer-supported reagent for these transformations under heterogeneous conditions is also described. These methods provide a fundamentally new approach to making small molecules containing the sulfonamide functional group. PMID- 14746464 TI - Detection of free nickel monocarbonyl, NiCO: rotational spectrum and structure. AB - Unsaturated transition metal carbonyls are important in processes such as organometallic synthesis, homogeneous catalysis, and photochemical decomposition of organometallics. In particular, a metal monocarbonyl offers a zeroth-order model for interpreting the chemisorption of a CO molecule on a metal surface in catalytic activation processes. Quite large numbers of theoretical papers have appeared which predict spectroscopic and structural properties of transition metal carbonyls. The nickel monocarbonyl NiCO has been one of the metal carbonyls most extensively studied by the theoretical calculations. At least 50 theoretical studies have been published on this simplest transition metal carbonyl up to the present time. However, experimental evidence of NiCO is much more sparse than theoretical predictions, and the actual structure of NiCO has never been determined by any experimental methods. This Communication reports the first preparation of free nickel monocarbonyl and observation of its rotational transitions. The NiCO molecule was generated by the sputtering reaction of a Ni cathode in the presence of CO. The accurate bond lengths of Ni-C and C-O were experimentally determined from isotopic data and were compared with the theoretical predictions for the first time. PMID- 14746465 TI - Total synthesis of spiruchostatin A, a potent histone deacetylase inhibitor. AB - The total synthesis of spiruchostatin A was accomplished, unambiguously confirming its structure. Key steps included the use of the Nagao thiazolidinethione auxiliary for a diastereoselective acetate aldol reaction and as an activated acylating agent for amide formation, and macrolactonization by the Yamaguchi protocol. Spiruchostatin A is shown to have biological activity similar to that of FK228, a potent histone deacetylase (HDAC) inhibitor in clinical trials. The spiruchostatin A analogue, epimeric at the beta-hydroxy acid, is inactive, highlighting the importance of stereochemistry at this position for interactions with HDACs. PMID- 14746466 TI - Non-covalent assembly of a photoswitchable surface. AB - A noncovalently bound multilayered thin film in which individual layers are linked by metal ligand interactions undergoes a photochemically initiated permanent change in surface wettability. The film consists of three separate layers: a SAM on gold of 4-[(10-mercaptodecyl)oxy]pyridine-2,6-dicarboxylic acid, a layer of Cu(II) ions that are deposited onto the SAM and bind symmetrically in the site provided by the two carboxylate groups and the pyridyl nitrogen atom, and a layer of cis-2,2'-dipyridylethylene, which caps the Cu(II) layer by complexation through both pyridyl nitrogen atoms (Film I). Photoexcitation of the film in chloroform at 300 nm leads to substantial cis-trans isomerization as indicated by conductivity, impedance, grazing incidence IR, and contact angle measurements. The latter show a decrease in contact angle (increase in wettability) of 17 degrees , which is attributed to exposure of both the underlying Cu(II) layer and one of the pyridyl ring nitrogen atoms following isomerization to the trans isomer. PMID- 14746467 TI - Switching of chiral induction in helical aromatic oligoamides using solid state solution state equilibrium. AB - The introduction of an R asymmetric center in an aromatic oligoamide that adopts stable helical conformations leads to a significant shift of the equilibrium between the right-handed and left-handed helices in solution: the R-P and R-M helices are diastereoisomers. However, these two species were found to cocrystallize in 1:1 proportions. Thus the chiral induction observed in solution is switched off in the solid state. This phenomenon represents an original and unexpected means to control handedness in helical oligomers. PMID- 14746468 TI - Extending the chemistry of the uranyl ion: Lewis acid coordination to a U=O oxygen. AB - Treatment of the thf adduct UO2(NCN)thf (NCN = [(Me3SiN)CPh(NSiMe3)]) (1) with 2 equiv of B(C6F5)3 provides UO{OB(C6F5)3}(NCN)2 (2) the first example of a neutral uranyl complex exhibiting Lewis basic behavior. The crystal structure of 2 shows a U=O-B interaction with an elongated U=O bond (1.898(3) A). Raman spectroscopy suggests weakening of the O=U=O bonding, giving the lowest reported symmetric stretching frequency for a monomeric uranyl complex, nu1 = 780 cm-1. The borane can be selectively removed using PMe3 to give the coordinatively unsaturated UO2(NCN)2 (3) or using tBuNC to provide UO2(CNBut)(NCN)2 (4), the first example of an isonitrile coordinated to uranium. PMID- 14746469 TI - Discovery of potent cell migration inhibitors through total synthesis: lessons from structure-activity studies of (+)-migrastatin. AB - Synthesis of highly active migrastatin-based tumor migration cell inhibitors has been accomplished. Our flexible and concise total synthesis of migrastatin has allowed us to explore otherwise inaccessible migrastatin-derived structural motifs. This effort has resulted in the discovery of analogues with tumor cell migration inhibitory activity 3 orders of magnitude higher than that of the natural product. PMID- 14746470 TI - Total synthesis and examination of three key analogues of ramoplanin: a lipoglycodepsipeptide with potent antibiotic activity. AB - The total synthesis and evaluation of three key ramoplanin aglycon analogues are detailed. The first (5a) represents replacement of the labile depsipeptide ester with a stable amide (HAsn2 --> Dap2) with removal of the HAsn pendant carboxamide, and it was found to be slightly more potent than the natural aglycon in antimicrobial assays providing a new lead structure with an improved profile and a more stable and accessible macrocyclic template on which to conduct structure-function studies. In contrast, a second amide analogue 5b which contains a single additional methylene relative to 5a (HAsn2 --> Dab2) was found to be inactive in antimicrobial assays (>100-fold loss in activity). The third key analogue 5c in which the Asn1 lipid side chain was replaced with an acetyl group revealed that it contributes significantly to the antimicrobial activity (16-fold) of the ramoplanins, but is not essential. PMID- 14746471 TI - Modular DNA-programmed assembly of linear and branched conjugated nanostructures. AB - A new strategy for self-assembly and covalent coupling of encoded molecular modules into nanostructures with predetermined connectivity has been developed. The method uses DNA-functionalized oligo(phenylene ethynylene)-derived organic modules for controlling the assembly and covalent coupling of multiple modules. Rigid linear modules (LM) and tripoidal modules (TM) were functionalized with short oligonucleotides at each terminus. They can hybridize and thereby link up modules containing complementary sequences. Each terminus of the oligo(phenylene ethynylene) modules also consists of a salicylaldehyde moiety, which can form metal-salen complexes with other modules. The salicylaldehyde groups of two modules are brought in proximity when their adjoining DNA sequences are complementary, and they selectively form a manganese-salen complex in the presence of ethylenediamine and manganese acetate. The resulting structures consist of a matrix of linear and branched oligo(phenylene ethynylene)s which are linked by conjugated and rigid manganese-salen complexes. These nanostructures are potential conductors for applications in molecular electronics. PMID- 14746472 TI - Dissecting the cholera toxin-ganglioside GM1 interaction by isothermal titration calorimetry. AB - The complex of cholera toxin and ganglioside GM1 is one of the highest affinity protein-carbohydrate interactions known. Herein, the GM1 pentasaccharide is dissected into smaller fragments to determine the contribution of each of the key monosaccharide residues to the overall binding affinity. Displacement isothermal titration calorimetry (ITC) has allowed the measurement of all of the key thermodynamic parameters for even the lowest affinity fragment ligands. Analysis of the standard free energy changes using Jencks' concept of intrinsic free energies reveals that the terminal galactose and sialic acid residues contribute 54% and 44% of the intrinsic binding energy, respectively, despite the latter ligand having little appreciable affinity for the toxin. This analysis also provides an estimate of 25.8 kJ mol(-1) for the loss of independent translational and rotational degrees of freedom on complexation and presents evidence for an alternative binding mode for ganglioside GM2. The high affinity and selectivity of the GM1-cholera toxin interaction originates principally from the conformational preorganization of the branched pentasaccharide rather than through the effect of cooperativity, which is also reinvestigated by ITC. PMID- 14746473 TI - Versatile protein biotinylation strategies for potential high-throughput proteomics. AB - We present intein-mediated approaches for efficient biotinylation of proteins site-specifically. The reactive C-terminal thioester generated from intein assisted protein splicing (either in vitro or in live cells) served as an attractive and exclusive site for attaching cysteine-containing biotin. Using these novel biotinylation strategies, we were able to efficiently biotinylate many proteins from different biological sources in a potentially high-throughput, high-content fashion. Some of these proteins were subsequently immobilized, in a very simple manner, onto different avidin-functionalized solid surfaces for applications such as protein microarray and surface plasmon resonance (SPR) spectroscopy, highlighting the numerous advantages of using biotin over other tags (e.g., GST, His-tag, etc.) as the method of choice in protein purification/immobilization. In addition, our intein-mediated strategies provided critical advantages over other protein biotinylation strategies in a number of ways. For the first time, we also successfully demonstrated that intein-mediated protein biotinylation proceeded adequately inside both bacterial and mammalian living cells, as well as in a cell-free protein synthesis system. Taken together, our results indicate the versatility of these intein-mediated strategies for potential high-throughput proteomics applications. They may also serve as useful tools for various biochemical and biophysical studies of proteins both in vitro and in vivo. PMID- 14746475 TI - Magneto-mechanical detection of nucleic acids and telomerase activity in cancer cells. AB - The ultra-sensitive magneto-mechanical detection of DNA, single-base-mismatches in nucleic acids, and the assay of telomerase activity are accomplished by monitoring the magnetically induced deflection of a cantilever functionalized with magnetic beads associated with the biosensing interface. The analyzed M13phi DNA hybridized with the nucleic acid-functionalized magnetic beads is replicated in the presence of dNTPs that include biotin-labeled dUTP. The resulting beads are attached to an avidin-coated cantilever, and the modified cantilever is deflected by an external magnetic field. Similarly, telomerization of nucleic acid-modified magnetic beads in the presence of dNTPs, biotin-labeled dUTP, and telomerase from cancer cell extracts and the subsequent association of the magnetic beads to the cantilever surface results in the lever deflection by an external magnetic field. M13phi DNA is sensed with a sensitivity limit of 7.1 x 10(-20) M by the magneto-mechanical detection method. PMID- 14746474 TI - Regioselective cis-trans isomerization of arachidonic double bonds by thiyl radicals: the influence of phospholipid supramolecular organization. AB - Trans unsaturated fatty acids in humans may be originated by two different contributions. The exogenous track is due to dietary supplementation of trans fats and the endogenous path deals with free-radical-catalyzed cis-trans isomerization of fatty acids. Arachidonic acid residue (5c,8c,11c,14c-20:4), which has only two out of the four double bonds deriving from the diet, was used to differentiate the two paths and to assess the importance of a radical reaction. A detailed study on the formation of trans phospholipids catalyzed by the HOCH2CH2S* radical was carried out on L-alpha-phosphatidylcholine from egg lecithin and 1-stearoyl-2-arachidonoyl-L-alpha-phosphatidylcholine (SAPC) in homogeneous solution or in large unilamellar vesicles (LUVET). Thiyl radicals were generated from the corresponding thiol by either gamma-irradiation or UV photolysis, and the reaction course was followed by GC, Ag/TLC, and 13C NMR analyses. The isomerization was found to be independent of cis double bond location (random process) in i-PrOH solution. In the case of vesicles, the supramolecular organization of lipids produced a dramatic change of the isomerization outcome: (i) in egg lecithin, the reactivity of arachidonate moieties is higher than that of oleate and linoleate residues, (ii) in the linoleate residues of egg lecithin, the 9t,12c-18:2 isomer prevailed on the 9c,12t-18:2 isomer (3:1 ratio), and (iii) a regioselective isomerization of SAPC arachidonate residues occurred in the 5 and 8 positions. This effect of "positional preference" indicates that thiyl radicals entering the hydrophobic region of the membrane bilayer start to isomerize polyunsaturated fatty acid residues having the double bonds nearest to the membrane surfaces. We propose that arachidonic acid and its trans isomers can function as biomarkers in membranes for distinguishing the two trans fatty acid-forming pathways. PMID- 14746476 TI - Quenched auto-ligating DNAs: multicolor identification of nucleic acids at single nucleotide resolution. AB - We describe the synthesis and study of multicolor quenched autoligating (QUAL) probes for identification and discrimination of closely related RNA and DNA sequences in solution and in bacteria. In these probes, a dabsyl quencher doubles as an activator in the oligonucleotide-joining reaction. The oligonucleotides remain dark until they bind at adjacent sites, and "light up" on nucleophilic displacement of the dabsyl probe by the phosphorothioate probe. Four fluorescent dye conjugates were prepared and tested with probes and targets that differ by one nucleotide. Experiments on polymer beads show clear color-based discrimination of DNAs added in solution. Two-color quenched probe pairs were then tested in the discrimination of 16S rRNA sequences in Escherichia coli. Single nucleotide resolution was achieved in the cells with green/red QUAL probes, allowing identification of a one-base sequencing error in the 16S rRNA database. Finally, QUAL probes were successfully applied in live bacterial cells. The method requires only incubation followed by fluorescence imaging, and requires no enzymes, added reagents, cross-linking, fixing, or washes. Because probes must bind side-by-side to generate signal, there is little or no interference from unintended protein binding, which can occur with other probe types. The results suggest that QUAL probes may be of general use in the detection and identification of sequences in solution, on microarrays, and in microorganisms. PMID- 14746477 TI - Influences of ribonucleotide on a duplex conformation and its thermal stability: study with the chimeric RNA-DNA strands. AB - To understand the influences of the ribonucleotide on a duplex conformation and its stability, we systematically studied the CD spectra and the thermodynamics of nucleic acid duplexes formed by the chimeric RNA-DNA strand in which ribonucleotides and deoxyribonucleotides were covalently attached. It was found that the duplex stability was context-dependent and independent of the number of ribonucleotides in the chimeric strand, whereas the CD spectra showed less overall structural perturbation by the chimeric junctions. Combining the results of the CD and the thermodynamic data revealed a stability-structure relationship for the duplexes. Importantly, DeltaG(o)37 values estimated for the chimeric junction formation in the RNA-DNA/DNA and the RNA-DNA/RNA duplexes were close to those of RNA/DNA and RNA/RNA interactions, respectively. Furthermore, DeltaG(o)37s of the DNA-RNA/DNA and DNA-RNA/DNA-RNA junctions were similar to those of the DNA duplex, and the values of DNA-RNA/RNA-DNA were similar to those of the DNA/RNA. The thermodynamic analyses suggest that the 5'-nucleotide may be the crucial factor that determines the stability at the chimeric junction. Our results not only suggest influences of the ribonucleotide on a duplex conformation and its stability but also are useful for the design of RNA-DNA chimeric strands applicable to biotechnology. PMID- 14746478 TI - Efficient trapping of HNO by deoxymyoglobin. AB - Nitrosyl hydride, HNO, also commonly termed nitroxyl, is a transient species that has been implicated in the biological activity of nitric oxide, NO. Herein, we report the first generation of a stable HNO-metal complex by direct trapping of free HNO. Deoxymyoglobin (Mb-Fe(II)) rapidly reacts with HNO produced from the decomposition of methylsulfonylhydroxylamine (MSHA) or Angeli's salt (AS) in aqueous solutions from pH 7 to pH 10, forming an adduct, Mb-HNO. The unique 1H NMR signal of the Fe-bound HNO at 14.8 ppm allows definitive proof of its formation. The generation of Mb-HNO and quantification of various myoglobin byproducts were accomplished by correlation of 1H NMR, UV-vis, and EPR spectroscopies. Typically, the maximum Mb-HNO yield obtained is 60-80%; competitive side reactions with byproducts as well as the further reactivity of the Mb-HNO decrease the overall yield. At pH 10, the observed rate of Mb-HNO generation by trapping HNO from MSHA is close to that for MSHA decomposition; kinetic simulations give a lower limit to the bimolecular rate of trapping as 1.4 x 10(4) M(-1) s(-1). The binding of HNO to deoxymyoglobin is rapid and essentially irreversible, which suggests that the biological activity of nitroxyl may be mediated by its reactivity with ferrous heme proteins such as myoglobin and hemoglobin. PMID- 14746479 TI - Toward a new genetic system with expanded dimensions: size-expanded analogues of deoxyadenosine and thymidine. AB - We describe the design, preparation, and properties of two key building blocks of a size-expanded genetic system. Nucleoside analogues of the natural nucleosides dA and dT are reported in which the fusion of a benzo ring increases their size by ca. 2.4 A. The expanded dA analogue (dxA), having a tricyclic base, was first reported by Leonard nearly three decades ago. We describe a shortened and more efficient approach to this compound. The expanded dT analogue (dxT), a methylquinazolinedione C-glycoside, was previously unknown; we describe its preparation in eight steps from 5-methylanthranilic acid. The key glycoside bond formation employed Pd-mediated coupling of an aryl iodide precursor with a dihydrofuran derivative of deoxyribose. Both nucleosides are shown to be efficient fluorophores, emitting light in the blue-violet range. The base protected phosphoramidite derivatives were prepared, and short oligonucleotides containing them were characterized. The two size-expanded nucleosides are key components of a new four-base genetic system designed to form helical paired structures having a diameter greater than that of natural DNA. Elements of the design of this expanded genetic molecule, termed xDNA, are discussed, including the possibility of up to eight base pairs of information storage capability. PMID- 14746480 TI - The photochemistry of polydonor-substituted phthalimides: Curtin-Hammett-type control of competing reactions of potentially interconverting zwitterionic biradical intermediates. AB - The results of studies designed to obtain information about the factors that control the chemical efficiencies/regioselectivities and quantum yields of single electron transfer (SET)-promoted reactions of acceptor-polydonor systems are reported. Photochemical and photophysical investigations were carried out with bis-donor tethered phthalimides and naphthalimides of general structure N phthalimido- and N-naphthalimido-CH2CH2-D-CH2CH2-NMsCH2-E (E = SiMe3 or CO2NBu4 and D = NMs, O, S, and NMe). These substrates contain common terminal donor groups (NMsCH2SiMe3 or NMsCH2CO2NBu4) that have known oxidation potentials and cation radical fragmentation rates. Oxidation potentials and fragmentation rates at the other donor site in each of these substrates are varied by incorporating different heteroatoms and/or substituents. Photoproduct distribution, reaction quantum yield, and fluorescence quantum yield measurements were made. The results show that photocyclization reactions of alpha-trimethylsilylmethansulfonamide (E = SiMe3)- and alpha-carboxymethansulfonamide (E = CO2NBu4)-terminated phthalimides and naphthalimides that contain internal sulfonamide, ether, and thioether donor sites (D = NMs, O, or S) are chemically efficient (80-100%) and that they take place exclusively by a pathway involving sequential photoinduced SET (zwitterionic biradical desilylation or decarboxylation) biradical cyclization. In contrast, photoreactions of alpha-trimethylsilylmethansulfonamide and alpha-carboxymethansulfonamide-terminated phthalimides and naphthalimides that that contain an internal tertiary amine donor site (D = NMe) are chemically inefficient and follow a pathway involving alpha-deprotonation at the tertiary amine radical cation center in intermediate, iminium radical-containing, zwitterionic biradicals. In addition, the quantum efficiencies for photoreactions of alpha-trimethylsilylmethansulfonamide- and alpha-carboxymethansulfonamide terminated phthalimides are dependent on the nature of the internal donor (eg., phi = 0.12 for D = NMs, E = SiMe3; phi = 0.02 for D = S, E = SiMe3; phi = 0.04 for D = NMe, E = SiMe3). The results of this effort are discussed in terms of how the relative energies of interconverting zwitterionic biradical intermediates and the energy barriers for their alpha-heterolytic fragmentation reactions influence the chemical yields and quantum efficiencies of SET promoted photocyclization reactions of acceptor-polydonor substrates. PMID- 14746481 TI - Charge separation in DNA via consecutive adenine hopping. AB - Charge transfer in DNA is of current interest because of the involvement of charge transfer in oxidative DNA damage and electronic molecular devices. We have investigated the charge separation process via the consecutive adenine (A) hopping mechanism using laser flash photolysis of DNA conjugated with naphthaldiimide (NDI) as an electron acceptor and phenothiazine (PTZ) as a donor. Upon the 355-nm laser flash excitation of NDI, the charge separation and recombination process between NDI and PTZ was observed. The yields of the charge separation via the consecutive A-hopping were slightly dependent upon the number of A bases between the two chromophores, while the charge recombination rate was strongly dependent upon the distance. The charge-separated state persisted over 300 micros when NDI was separated from PTZ by eight A bases. Furthermore, the rate constant of the A-hopping process was determined to be 2 x 10(10) s(-1) from an analysis of the yield of the charge separation depending on the number of A hopping steps. PMID- 14746482 TI - Preparation of oligodiazo compounds by using the suzuki coupling reaction and characterization of their photoproducts. AB - [9-[10-(4-tert-Butyl-2,6-dimethyl)phenyl]anthryl](4-bromo-2,6 dimethylphenyl)diazomethane was found to be stable enough to survive under Suzuki coupling conditions and underwent mono-, di-, and trisubstitution with benzene mono-, di-, and triboronic acids to afford benzene derivatives having one, two, and three diazo units, respectively. The products from irradiation of those diazo compounds were characterized by ESR and SQUID measurements, which revealed that triplet, quintet, and septet ground states were formed from mono-, bis-, and tris(diazo) compounds, respectively. The stability of those high-spin species was estimated by temperature-dependent ESR and UV/vis measurements as well as laser flash photolysis, which indicated that all three species are stable up to 160 K and have a half-life of a few seconds in solution at room temperature. The finding unequivocally shows that a precursor diazo unit can basically be handled as a building block to construct ploydiazo compounds and that persistent triplet carbenes, even though they greatly lose typical reactivity as a triplet carbene, still retain electronic properties and act as a spin source when aligned properly in the pi-electron frameworks to generate a high-spin molecule with remarkable thermal stability. PMID- 14746483 TI - Chemical synthesis of selenium-modified oligoribonucleotides and their enzymatic ligation leading to an U6 SnRNA stem-loop segment. AB - The derivatization of nucleic acids with selenium is highly promising to facilitate nucleic acids structure determination by X-ray crystallography using the multiwavelength anomalous dispersion (MAD) technique. The foundation for such an approach has been laid by Huang, Egli, and co-workers and was exemplified on small DNA duplexes. Here, we present a comprehensive study on the preparation of RNAs containing 2'-Se-methylpyrimidine nucleoside labels. This includes the synthesis of a novel 2'-Se-methylcytidine phosphoramidite 11 and its incorporation into oligoribonucleotides by solid-phase synthesis. Deprotection of the oligonucleotides is achieved in the presence of millimolar amounts of threo 1,4-dimercapto-2,3-butandiol (DTT). With this additive, oxidation products and follow-up side-products are suppressed and acceptable HPLC traces of the crude material are obtained, so far tested for sequences of up to 22-mers. Moreover, an extensive investigation on the enzymatic ligation of the selenium-containing oligoribonucleotides demonstrates the high flexibility of the selenium approach. Our target sequence, an U6 snRNA stem-loop motif comprising all naturally occurring nucleoside modifications beside the Se-label is achieved by ligation using T4 RNA ligase. PMID- 14746484 TI - (2,7-Disubstituted-1,8-biphenylenedioxy)bis(dimethylaluminum) as bidentate organoaluminum Lewis acids: elucidation and synthetic Utility of the double electrophilic activation phenomenon. AB - A series of (2,7-disubstituted-1,8-biphenylenedioxy)bis(dimethylaluminum) (2) has been readily prepared in situ by treatment of the requisite 2,7-disubstituted-1,8 biphenylenediol (1) with Me3Al (2 equiv) in CH2Cl2 at room temperature; this primarily relies on the successful establishment of a new synthetic procedure of 1 starting from inexpensive m-anisidine. Evaluation of 2 as a bidentate organoaluminum Lewis acid has been performed by the reduction of ketonic substrates using Bu3SnH as a hydride source in comparison to the conventional monodentate Lewis acid dimethylaluminum 2,6-xylenoxide (11), uncovering the significantly high activation ability of 2 toward carbonyl. Particularly, (2,7 dimethyl-1,8-biphenylenedioxy)bis(dimethylaluminum) (2a) exerted the highest reactivity, which has also been emphasized in the Mukaiyama aldol reaction. The structure of the bidentate Lewis acid 2 was unambiguously determined by single crystal X-ray diffraction analysis of 2g possessing a bulky 3,5-di-tert butylphenyl substituent, revealing the rigid dimeric assembly in the solid state. The double electrophilic activation of carbonyl substrate by 2a has been supported by low-temperature 13C NMR analysis as well as theoretical study using the Gaussian 98 program. Moreover, unique stereoselectivity has been observed in the 2a-promoted Mukaiyama Michael addition, and highly chemoselective functionalization of carbonyl compounds in the presence of their acetal counterparts has been realized using 2a. Finally, the effectiveness of 2a for the activation of ether functionality has been demonstrated in the Claisen rearrangement of allyl vinyl ethers. PMID- 14746485 TI - Directed metalation route to ferroelectric liquid crystals with a chiral fluorenol core: the effect of restricted rotation on polar order. AB - A new series of smectic C* (SmC*) mesogens containing a chiral (R)-2-octyloxy side chain and either a fluorenone (2a-e) or chiral fluorenol (3a-e) core were synthesized using a combined directed ortho metalation-directed remote metalation strategy. The SmC phase formed by the fluorenol mesogens is more stable and has a wider temperature range than that formed by the fluorenone mesogens, which may be ascribed to intermolecular hydrogen bonding according to variable-temperature FT IR measurements. The C11 fluorenol mesogens (R,R)-3d and (S,R)-3d were obtained in diastereomerically pure form and gave reduced polarization (Po) values of +106 and +183 nC/cm2, respectively, at 10 K below the SmA*-SmC* phase transition temperature. The difference in Po values suggests that the chiral fluorenol core contributes to the spontaneous polarization of the SmC* phase. This is ascribed to the bent shape of the fluorenol core, which should restrict its rotation with respect to the side chains in the SmC* phase and favor one orientation of its transverse dipole moment along the polar axis, and to steric coupling of the core to the chiral 2-octyloxy side chain. PMID- 14746486 TI - Selective growth and distribution of crystalline enantiomers in hydrogels. AB - The crystallization of sodium chlorate (NaClO3) is a classic example of spontaneous chirality, since it is achiral in solution but adopts a chiral form in the solid state. While crystal growth of NaClO3 from pure aqueous solutions yields a 50:50 statistical distribution of d- and l-crystals, large enantiomeric excesses of either d- and l-crystals can be achieved by crystal growth in agarose gel, a naturally occurring chiral polysaccharide. The influence of gel density (0.1-0.75 wt %), temperature, and the diffusion of cosolvents on crystal distribution was discerned from statistical data obtained from 752 gel-mediated crystallization experiments yielding 12,384 individual crystals. These studies demonstrate that the magnitude and direction of the bias can be selectively engineered toward either d- or l-forms by changing the gelation conditions. Aqueous agarose gels infused with 48 wt % NaClO3 at 6 degrees C, favored the growth of d-NaClO3 crystals, with ee's reaching 22% at the highest gel concentrations. Crystal growth under methanol diffusion favored deposition of the opposite enantiomorph, l-NaClO3. The bias in the crystal distribution is enhanced at higher temperatures. Aqueous gels at 24 degrees C infused with methanol cosolvent favored l-NaClO3, with ee's reaching 53%. The changing magnitude and direction of the enantiomorph bias can be ascribed to differences in the agarose conformation and intermolecular interactions between the gel and crystal surfaces that inhibit the formation of the two enantiomers to different extents. PMID- 14746487 TI - Demonstrating the feasibility of monitoring the molecular-level structures of moving polymer/silane interfaces during silane diffusion using SFG. AB - In this paper, the feasibility of monitoring molecular structures at a moving polymer/liquid interface by sum frequency generation (SFG) vibrational spectroscopy has been demonstrated. N-(2-Aminoethyl)-3 aminopropyltrimethoxysilane (AATM, NH2(CH2)2NH(CH2)3Si(OCH3)3) has been brought into contact with a deuterated poly(methyl methacrylate) (d-PMMA) film, and the interfacial silane structure has been monitored using SFG. Upon initial contact, the SFG spectra can be detected, but as time progresses, the spectral intensity changes and finally disappears. Additional experiments indicate that these silane molecules can diffuse into the polymer film and the detected SFG signals are actually from the moving polymer/silane interface. Our results show that the molecular order of the polymer/silane interface exists during the entire diffusion process and is lost when the silane molecules traverse through the thickness of the d-PMMA film. The loss of the SFG signal is due to the formation of a new disordered substrate/silane interface, which contributes no detectable SFG signal. The kinetics of the diffusion of the silane into the polymer have been deduced from the time-dependent SFG signals detected from the AATM molecules as they diffuse through polymer films of different thickness. PMID- 14746488 TI - In situ TA-MS study of the six-membered-ring-based growth of carbon nanotubes with benzene precursor. AB - By using the in situ thermal analysis-mass spectroscopic technique, combined with transmission electron microscopic characterization of the carbon nanotube (CNT) product, we have studied the chemical vapor deposition (CVD) growth of CNTs with Fe-Co/gamma-Al2O3 catalyst and benzene precursor in the range of room temperature to 700 degrees C. The growth process has been clearly illuminated, which starts from the reduction of catalyst around 645 degrees C followed by the dissociation of carbon-hydrogen bonds of benzene and the sequential growth of CNTs. A surprising fact is that no possible hydrocarbon species derived from benzene was detected, indicating that the carbon-carbon bond was not broken under our experimental conditions. All of the experimental results strongly reinforce the six-membered-ring-based growth model, and a schematic elucidation is presented accordingly. This in situ study not only reveals the unique and convincing information directly related to the growth mechanism from the involved chemistry, but also provides a powerful way to clarify the mechanism of CVD synthesis of CNTs with other precursors. PMID- 14746490 TI - Solution-phase synthesis of single-crystalline iron phosphide nanorods/nanowires. AB - A solution-phase route for the preparation of single-crystalline iron phosphide nanorods and nanowires is reported. We have shown that the mixture of trioctylphosphine oxide (TOPO) and trioctylphosphine (TOP), which are commonly used as the solvents for semiconductor nanocrystal synthesis, is not entirely inert. In the current process, TOP, serving as phosphor source, reacts with Fe precursors to form FeP nanostructures with large aspect ratios. In addition, the experimental results show that both TOP and TOPO are necessary for the formation of FeP nanowires and their ratio appears to control the morphology of the produced FeP structures. A possible growth mechanism is discussed. PMID- 14746489 TI - Synthesis, structure, theoretical studies, and Ligand exchange reactions of monomeric, T-shaped arylpalladium(II) halide complexes with an additional, weak agostic interaction. AB - A series of monomeric arylpalladium(II) complexes LPd(Ph)X (L = 1-AdPtBu2, PtBu3, or Ph5FcPtBu2 (Q-phos); X = Br, I, OTf) containing a single phosphine ligand have been prepared. Oxidative addition of aryl bromide or aryl iodide to bis-ligated palladium(0) complexes of bulky, trialkylphosphines or to Pd(dba)2 (dba = dibenzylidene acetone) in the presence of 1 equiv of phosphine produced the corresponding arylpalladium(II) complexes in good yields. In contrast, oxidative addition of phenyl chloride to the bis-ligated palladium(0) complexes did not produce arylpalladium(II) complexes. The oxidative addition of phenyl triflate to PdL2 (L = 1-AdPtBu2, PtBu3, or Q-phos) also did not form arylpalladium(II) complexes. The reaction of silver triflate with (1-AdPtBu2)Pd(Ph)Br furnished the corresponding arylpalladium(II) triflate in good yield. The oxidative addition of phenyl bromide and iodide to Pd(Q-phos)2 was faster than oxidative addition to Pd(1-AdPtBu2)2 or Pd(PtBu3)2. Several of the arylpalladium complexes were characterized by X-ray diffraction. All of the arylpalladium(II) complexes are T shaped monomers. The phenyl ligand, which has the largest trans influence, is located trans to the open coordination site. The complexes appear to be stabilized by a weak agostic interaction of the metal with a ligand C-H bond positioned at the fourth-coordination site of the palladium center. The strength of the Pd.H bond, as assessed by tools of density functional theory, depended upon the donating properties of the ancillary ligands on palladium. PMID- 14746491 TI - Oxidation of CO on gold supported catalysts prepared by laser vaporization: direct evidence of support contribution. AB - Gamma-Al2O3, ZrO2, and TiO2 gold supported model catalysts have been synthesized by laser vaporization. Structural characterization using Transmission Electron Microscopy and X-ray Photoelectron Spectroscopy experiments have shown that the gold clusters deposited on the different supports have similar distribution of size centered around 3 nm and are in the metallic state. However, X-ray photoemission measurements also indicate lower binding energies than the usual Au 4f(7/2) at 84.0 eV for both alumina and titania supported catalysts, indicating a modification of the electronic structure of the metal. One has taken benefit of these features to study the influence of the nature of the support toward CO oxidation activities without being hindered by particle size or gold oxidic species effects. By comparing the activities of the different catalysts, it is concluded that the nature of the support directly affects the activity of gold. The following tendency is observed: titania and zirconia are superior to alumina as supports, titania being slightly better than zirconia. From XPS and activity results we can conclude that the existence of negatively charged clusters is not the key point to explain the high activity observed for Au/ZrO2 and Au/TiO2 catalysts and also that metallic Au is the major catalytically active phase. Hence, due to their very nature, titania and to a less extent zirconia should participate to the catalytic process. PMID- 14746492 TI - Water molecule adsorption on protonated dipeptides. AB - Equilibrium constants for the adsorption of the first water molecule on six protonated dipeptides (Gly-Gly+H(+), Gly-Ala+H(+), Ala-Gly+H(+), Ala-Ala+H(+), Pro-Gly+H(+), and Gly-Trp+H(+)) have been measured as a function of temperature, and DeltaH(o) and DeltaS(o) determined. Density functional theory calculations were performed for both the unsolvated peptides and the peptide water complexes at the B3LYP/6-311++G level. MP2/6-311++G** calculations were also carried out for Gly/Ala peptides. The calculations suggest that adsorption of a water molecule by these simple dipeptides is a complex process, both the unsolvated peptide and the peptide-water complexes have multiple conformations with similar free energies. Average DeltaH(o) and DeltaS(o) values derived from the calculations are in reasonable agreement with the experimental results. According to the calculations, the dominant water adsorption process involves a significant conformational change to accommodate a bridging water molecule. DeltaH(o) is diminished for Pro-Gly+H(+) mainly because the water interacts with a secondary amine, whereas for Gly-Trp+H(+), DeltaH(o) is significantly decreased by the loss of cation-pi interactions upon water adsorption. For unsolvated peptides the proton affinities of the N-terminus and the backbone carbonyl groups are known to be similar. Addition of a single water molecule causes a significant stabilization of the N-terminus protonation site. PMID- 14746493 TI - Heterotopic assemblage of two different disk-shaped ligands through trinuclear silver(I) complexation: ligand exchange-driven molecular motion. AB - The sandwich-shaped heterotopic trinuclear Ag+ complex Ag(3)1.2 was exclusively formed from two different tris(thiazolyl) and hexa(thiazolyl) disk-shaped ligands, 1 and 2, with the aid of three Ag+ ions. The variable-temperature 1H NMR study on its complexation behavior revealed that metal-ligand exchanges between the two neighboring thiazolyl nitrogen donors of 2 take place at the three Ag+ centers in concert. DeltaH++ and DeltaS++ for the exchange process were calculated to be 50.5 kJ mol(-1) and -26.7 J mol(-1) K(-1), respectively, and its energy barrier at 298 K was estimated to be 58.5 kJ mol(-1). Each concerted metal ligand exchange leads to an intramolecular 60 degrees-rotational motion ((P) <==>(M) conversion) between the two disk-shaped ligands. PMID- 14746494 TI - Electron density distribution of an oxamato bridged Mn(II)-Cu(II) bimetallic chain and correlation to magnetic properties. AB - The electron density distribution of the ferrimagnetic MnCu(pba)(H2O)3.2H2O chain compound, where pba stands for 1,3-propylenebis(oxamato), has been derived from high resolution X-ray diffraction measurements at 114 K using a multipolar model. The analysis of the chemical bonding has been carried out through the "Atoms in Molecules" formalism and thoroughly interpreted with regards to the strong intrachain and weak interchain magnetic couplings. The topological properties of the electron density on the oxamato bridge indicate large electron delocalization and conjugation effects, in addition to high charge transfer from both metals to the bridge. The resulting positive charges on Mn (+1.45 e) and Cu (+1.56 e) induce charge polarization of the bridge, leading to a shift of electron density from the central C atoms to the metal coordinating O and N atoms. The Mn-bridge interactions are mainly closed-shell interactions with low electron density at the corresponding bond critical points, whereas the Cu-bridge interactions exhibit significant covalent character. The Cu-N bonds are moreover stronger than the Cu-O bonds. The 3d Cu and Mn orbital populations are consistent with pyramidal and regular octahedral environments, respectively, in agreement with the loss of degeneracy due to ligand field effects. Interchain interaction pathways are evidenced by the existence of four bond critical points in hydrogen bond regions. Finally, these intrachain and interchain bonding features are correlated to the results of experimental and theoretical spin density distributions, as well as magnetic measurements. PMID- 14746495 TI - Scanning tunneling microscopy of self-assembled phenylene ethynylene oligomers on Au(111) substrates. AB - In this paper, we report the self-assembly, electrical characterization, and surface modification of dithiolated phenylene-ethynylene oligomer monolayers on a Au(111) surface. The self-assembly was accomplished by thiol bonding the molecules from solution to a Au(111) surface. We have confirmed the formation of self-assembled monolayers by scanning tunneling microscopy (STM) and optical ellipsometry, and have studied the kinetics of film growth. We suggest that self assembled phenylene ethynylene oligomers on Au(111) surfaces grow as thiols rather than as thiolates. Using low-temperature STM, we collected local current voltage spectra showing negative differential resistance at 6 K. PMID- 14746496 TI - Microwave spectra and structures of KrAuF, KrAgF, and KrAgBr; 83Kr nuclear quadrupole coupling and the nature of noble gas-noble metal halide bonding. AB - Microwave spectra of the complexes KrAuF and KrAgBr have been measured for the first time using a cavity pulsed jet Fourier transform microwave spectrometer. The samples were prepared by laser ablation of the metal from its solid and allowing the resulting plasma to react with an appropriate precursor (Kr, plus SF6 or Br2) contained in the backing gas of the jet (usually Ar). Rotational constants; geometries; centrifugal distortion constants; vibration frequencies; and 197Au, 79Br, and 81Br nuclear quadrupole coupling constants have all been evaluated. The complexes are unusually rigid and have short Kr-Au and Kr-Ag bonds. The 197Au nuclear quadrupole coupling constant differs radically from its value in an AuF monomer. In addition 83Kr hyperfine structure has been measured for KrAuF and the previously reported complex KrAgF. The geometry of the latter has been reevaluated. Large values for the 83Kr nuclear quadrupole coupling constants have been found for both complexes. Both the 197Au and 83Kr hyperfine constants indicate a large reorganization of the electron distribution on complex formation. A thorough assessment of the nature of the noble gas-noble metal bonding in these and related complexes (NgMX; Ng is a noble gas, M is a noble metal, and X is a halogen) has been carried out. The bond lengths are compared with sums of standard atomic and ionic radii. Ab initio calculations have produced dissociation energies along with Mulliken populations and other data on the electron distributions in the complexes. The origins of the rigidity, dissociation energies, and nuclear quadrupole coupling constants are considered. It is concluded that there is strong evidence for weak noble gas-noble metal chemical bonding in the complexes. PMID- 14746497 TI - Mapping the electronic structure of the blue copper site in plastocyanin by NMR relaxation. AB - The biological function of metalloproteins stems from the electronic and geometric structures of their active sites. Thus, in blue copper proteins such as plastocyanins, an unusual electronic structure of the metal site is believed to contribute to the rapid, long-range electron-transfer reactivity that characterizes these proteins. To clarify this structure-function relationship, numerous quantum chemical calculations of the electronic structure of the blue copper proteins have been made. However, the obtained structures depend strongly on the applied model. Experimental approaches based on ENDOR spectroscopy and X ray absorption have also been used to elucidate the electronic structure of the blue copper site. Still, the determination of the electronic structure relies on a calibration with quantum chemical calculations, performed on small model complexes. Here we present an approach that allows a direct experimental mapping of the electron spin delocalization in paramagnetic metalloproteins using oxidized plastocyanin from Anabaena variabilis as an example. The approach utilizes the longitudinal paramagnetic relaxation of protons close to the metal site and relies on the dependence of these relaxations on the spatial distribution of the unpaired electron of the metal ion. Surprisingly it is found that the unpaired electron of the copper ion in plastocyanin is less delocalized than predicted by most of the quantum chemical calculations. PMID- 14746498 TI - Role of the spacer in the singlet-singlet energy transfer mechanism (Forster vs Dexter) in cofacial bisporphyrins. AB - The cofacial bisporphyrins H4DPS (DPS = 4,6-bis[5-(2,8,13,17-tetraethyl-3,7,12,18 tetramethylporphyrinyl)]dibenzothiophene), H4DPO (DPO = 4,6-bis[5-(2,8,13,17 tetraethyl-3,7,12,18-tetramethylporphyrinyl)]dibenzofuran), H4DPX (DPX = 4,5 bis[5-(2,8,13,17-tetraethyl-3,7,12,18-tetramethylporphyrinyl)]-9,9 dimethylxanthene), H4DPA (DPA = 1,8-bis[5-(2,8,13,17-tetraethyl-3,7,12,18 tetramethylporphyrinyl)]anthracene), and H4DPB (DPB = 1,8-bis[5-(2,8,13,17 tetraethyl-3,7,12,18-tetramethylporphyrinyl)]biphenylene) have been monometalated by Zn(OAc)2.2H2O and by GaCl3 to explore the singlet-singlet energy transfer from the photoexcited metal porphyrin center to the linked free base porphyrin. The spectroscopic (UV-vis and fluorescence) and photophysical properties (fluorescence lifetimes, tauF, and quantum yields, phiF) have been investigated at 298 and 77 K in degassed 2-MeTHF for the donor-acceptor systems, (Zn)H2DPS, (Zn)H2DPO, (Zn)H2DPA, (Zn)H2DPX, and (Zn)H2DPB, as well as for the bis-zinc complexes, (Zn)2DPS, (Zn)2DPO, (Zn)2DPX, and (Zn)2DPB, respectively, and the monoporphyrin derivatives, H2P, (Zn)P, and (Ga-OMe)P (P2- = 5-phenyl-2,8,13,17 tetraethyl-3,7,12,18-tetramethylporphyrin-dianion). The singlet-singlet energy transfer rate constants (KET) were obtained using KET = (1/tauF -1/tauFo), where tauFo is the fluorescence lifetime of the corresponding bis-zinc(II) systems (or (Zn)P and (Ga-OMe)P) where no energy transfer occurs. The tauF value for three bis-zinc(II) compounds varies from 1.69 to 2.01 ns and is 1.84 (at 298 K) and 3.20 ns (at 77 K) for (Ga-OMe)P. In the donor-acceptor bismacrocycles, depending on the spacer and the temperature, the fluorescence lifetimes decrease down to 50 240 ps. The KET values range from approximately 4 to approximately 21 (ns(-1)) and have been analyzed considering both the Forster and the Dexter mechanisms. Using the C(meso)-C(meso) distance parameters in the calculations, the Forster and Dexter mechanisms operate for DPS and DPO, and for DPA, DPX, and DPB spacer systems, respectively. The limit distance where one mechanism dominates over the other is estimated to be around 5-6 A. PMID- 14746499 TI - Real-time 2D NMR identification of analytes undergoing continuous chromatographic separation. AB - We have recently proposed and demonstrated an approach that enables the acquisition of multidimensional nuclear magnetic resonance (NMR) spectra within a single scan. A promising application opened up by this new accelerated form of data acquisition concerns the possibility of monitoring in real time the chemical nature of analytes subject to a continuous flow. The present paper illustrates such potential, with the real-time acquisition of a series of 2D 1H NMR spectra arising from a mixture of compounds subject to a continuous liquid chromatography (LC) separation. This real-time 2D NMR identification of chemicals eluted minutes apart under usual LC-NMR conditions differs from the way in which LC-2D NMR has hitherto been carried out, which relies on stopped-flow modes of operations whereby fractions are first collected and then subject to individual, aliquot-by aliquot analyses. The real-time LC-2D NMR experiment hereby introduced can be implemented in a straightforward manner using modern commercial LC-NMR hardware, thus opening up immediate possibilities in high-throughput characterizations of complex molecules. PMID- 14746500 TI - Quasi-relativistic density functional study of aurophilic interactions. AB - Fifteen molecules containing the Au(I) species have been calculated by ab initio HF and MP2 methods and by five different density functional approaches. The aurophilic Au(d10)-Au(d10) bonding mechanism has been investigated. Both, one electron interactions (i.e., electrostatic, polarization, charge transfer, and orbital interference) and two-electron effects (i.e., correlation, dispersion) contribute significantly to the so-called 'secondary' or metallophilic bonds representing the Au-Au interaction. Second, the applicability of density functional approaches to this type of bonding has been tested. It is well-known that present day density functionals are not yet designed to simulate the long range London dispersion forces between nonoverlapping systems, whereas they approximately reproduce the short range dynamical electron correlations of strongly overlapping chemically bonded nondegenerate species. It is found here empirically for the investigated groups of gold(I) cluster compounds that simple local density functionals (LDF) of the Slater (or Slater plus Vosko) type yield rather reasonable estimates for the equilibrium distances, and (on the average) also for the aurophilic interaction energies, though with rather large standard deviations. Still LDF are useful for survey investigations of Au cluster compounds. Common gradient corrected DF are not recommended here, nor are the large core pseudopotentials for Au. PMID- 14746501 TI - Photoinduced intramolecular charge transfer in 4-(dimethyl)aminobenzonitrile--a theoretical perspective. AB - Recent advances in time-dependent density functional theory (TDDFT) have led to computational methods that can predict properties of photoexcited molecules with satisfactory accuracy at comparably moderate cost. We apply these methods to study the photophysics and photochemistry of 4-(dimethyl)aminobenzonitrile (DMABN). DMABN is considered the paradigm of photoinduced intramolecular charge transfer (ICT), leading to dual fluorescence in polar solvents. By comparison of calculated emission energies, dipole moments, and vibrational frequencies with recent results from transient spectroscopy measurements, a definitive assignment of the electronic and geometric structure of the two lowest singlet excited states of DMABN is possible for the first time. We investigate the mechanism of the ICT reaction by means of minimum energy path calculations. The results confirm existing state-crossing models of dual fluorescence. Our study suggests that analytical TDDFT derivative methods will be useful to predict and classify emissive properties of other donor-acceptor systems as well. PMID- 14746502 TI - Hydration structure and free energy of biomolecularly specific aqueous dications, including Zn2+ and first transition row metals. AB - The hydration of some of the alkaline earth divalent metal cations and first row transition metal cations is considered within the quasi-chemical theory of solutions. Quantum chemical calculations provide information on the chemically important interactions between the ion and its first-shell water molecules. A dielectric continuum model supplies the outer-shell contribution. The theory then provides the framework to mesh these quantities together. The agreement between the calculated and experimental quantities is good. For the transition metal cations, it is seen that the ligand field contributions play an important role in the physics of hydration. Removing these bonding contributions from the computed hydration free energy results in a linear decrease in the hydration free energy along the period. It is precisely such effects that molecular mechanics force fields have not captured. The implications and extensions of this study to metal atoms in proteins are suggested. PMID- 14746503 TI - Primary intermediates of oxygen photoevolution reaction on TiO2 (Rutile) particles, revealed by in situ FTIR absorption and photoluminescence measurements. AB - Primary intermediates of oxygen photoevolution (water photooxidation) reaction at the TiO2 (rutile)/aqueous solution interface were investigated by in situ multiple internal reflection infrared (MIRIR) absorption and photoluminescence (PL) measurements. UV irradiation of TiO2 in the presence of 10 mM Fe3+ in the solution caused the appearance of a new peak at 838 cm(-1) and a shoulder at 812 cm(-1). Detailed investigations of the effects of solution pH, the presence of methanol as a hole scavenger, and isotope exchange in water (H2(16)O-->H2(18)O) on the spectra have shown that the 838- and 812-cm(-1) bands can be assigned to the O-O stretching mode of surface TiOOH and TiOOTi, respectively, produced as primary intermediates of the oxygen photoevolution reaction. The results give strong support to our previously proposed mechanism that the oxygen photoevolution is initiated by a nucleophilic attack of a H2O molecule on a photogenerated hole at a surface lattice O site, not by oxidation of surface OH group by the hole. The conclusion is supported by PL measurements. A plausible reaction scheme is proposed for the oxygen photoevolution on TiO2 (rutile) in aqueous solutions of pH less than about 12. PMID- 14746504 TI - Protein kinase C inhibits formation of va gene transcription initiation complex. AB - Activation of protein kinase c (PKC) reduces transcription from the polymerase III (pol III)-transcribed adenovirus VA gene. Data presented here support a role for PKC in disrupting the formation of transcription-competent initiation complexes. The study used the plasmids VA and VA/EL (VA gene with a linker to distinguish its transcript from that of the VA gene) in in vitro assays to show that preincubation of either template for a minimum of 10 min before the activation of PKC did not result in PKC-induced repression of transcription. In contrast, under the same conditions, efficient transcription occurs from a preincubated template but not from a second template if it is added during or after the activation of PKC. Simultaneous preincubation of both VA and VA/EL resulted in efficient transcription from both templates. Rescue experiments confirm that PKC modifies a target within transcription factor B (TFIIIB) because phosphocellulose fractionation of whole-cell extracts that yield partially purified pol III transcription factor, TFIIIB, successfully rescues VA transcription from PKC-induced repression. Subsequent studies confirmed that the TATA box-binding protein (TBP), a constituent of TFIIIB, substituted for the crude preparation of TFIIIB. These data support a conclusion that activation of PKC triggers a cascade that likely involves the sequestration or degradation of TBP, resulting in the disruption of the steps that leads to successful pol III transcription initiation. PMID- 14746506 TI - The impact of the completed human genome sequence on the development of novel therapeutics for human disease. AB - With the official completion of the Human Genome Project in April 2003, we have both the opportunity and the imperative to translate this unprecedented scientific accomplishment into tangible improvements in human health. Medical benefits from the genome will come in stages and can be conceptualized as occurring in three areas: improved understanding of disease causation at the molecular level, improved diagnosis and disease classification based on genetic profiles, and new therapeutics based on targets identified in the genome. These improvements will require increased physician understanding of genetic principles applied to common diseases. PMID- 14746507 TI - Toward Alzheimer therapies based on genetic knowledge. AB - Genetic analysis has allowed the dissection of the pathogenic pathway that leads to Alzheimer's disease. It has also been integral to the development of earlier and more accurate diagnostic practices. This analysis has identified many potential therapeutic targets, and clinical trials aimed at these targets are now under way. If these approaches are successful, it will be a spectacular validation of genetic-knowledge-based treatment strategies; if they are not, researchers will need to re-evaluate this approach toward understanding and developing strategies for treating complex diseases. PMID- 14746508 TI - Inherited diseases involving g proteins and g protein-coupled receptors. AB - Heterotrimeric G proteins couple seven-transmembrane receptors for diverse extracellular signals to effectors that generate intracellular signals altering cell function. Mutations in the gene encoding the alpha subunit of the G protein coupling receptors to stimulation of adenylyl cyclase cause developmental abnormalities of bone, as well as hormone resistance (pseudohypoparathyroidism caused by loss-of-function mutations) and hormone hypersecretion (McCune-Albright syndrome caused by gain-of-function mutations). Loss- and gain-of-function mutations in genes encoding G protein-coupled receptors (GPCRs) have been identified as the cause of an increasing number of retinal, endocrine, metabolic, and developmental disorders. GPCRs comprise an evolutionarily conserved gene superfamily ( 1 ). By coupling to heterotrimeric G proteins, GPCRs transduce a wide variety of extracellular signals including monoamine, amino acid, and nucleoside neurotransmitters, as well as photons, chemical odorants, divalent cations, hormones, lipids, peptides and proteins. Following a brief overview of G protein-coupled signal transduction, we review the growing body of evidence that mutations in genes encoding GPCRs and G proteins are an important cause of human disease. PMID- 14746509 TI - The scientific basis for the current treatment of Parkinson's disease. AB - Parkinson's disease (PD) is an age-related neurodegenerative disease that affects approximately one million people in the United States. The introduction of levodopa revolutionized the treatment for this disorder, but the long-term utility of the drug is limited by motor complications, the development of features such as postural instability and dementia that do not respond to treatment, and continued disease progression. Insights into the organization of the basal ganglia in the normal and PD conditions has permitted the design of new treatment strategies that reduce the risk of developing motor complications. Additionally, increased knowledge of the mechanisms responsible for cell death in PD has permitted the development of putative neuroprotective drugs that might slow or stop disease progression. No drug has yet been established to alter the rate of disease progression, but the rapid pace of research offers reason for optimism. PMID- 14746510 TI - Progress in antisense technology. AB - Antisense technology exploits oligonucleotide analogs to bind to target RNAs via Watson-Crick hybridization. Once bound, the antisense agent either disables or induces the degradation of the target RNA. Antisense agents can also alter splicing. During the past decade, much has been learned about the basic mechanisms of antisense, the medicinal chemistry, and the pharmacologic, pharmacokinetic, and toxicologic properties of antisense molecules. Antisense technology has proven valuable in gene functionalization and target validation. With one drug marketed, Vitravenetm, and approximately 20 antisense drugs in clinical development, it appears that antisense drugs may prove important in the treatment of a wide range of diseases. PMID- 14746511 TI - Serum proteomics in cancer diagnosis and management. AB - Mass spectrometry-based diagnostics has the potential to revolutionize molecular medicine. Using modern mass-spectrometer technologies, clinical tests can be developed that are practical, robust, accurate, and inexpensive. Serum proteomic pattern profiling couples mass spectrometry with adaptive artificial-intelligence based bioinformatics, which can now be employed to detect pathological states reflected in the serum proteome. With this approach, rapid and cost-effective tests with exquisite clinical sensitivity and specificity are emerging. These tools may dramatically change how disease is detected, monitored, and managed. PMID- 14746512 TI - Molecular neurobiology of drug addiction. AB - Addiction can be viewed as a form of drug-induced neural plasticity. One of the best-established molecular mechanisms of addiction is upregulation of the cAMP second messenger pathway, which occurs in many neuronal cell types in response to chronic administration of opiates or other drugs of abuse. This upregulation and the resulting activation of the transcription factor CREB appear to mediate aspects of tolerance and dependence. In contrast, induction of another transcription factor, termed DeltaFosB, exerts the opposite effect and may contribute to sensitized responses to drug exposure. Knowledge of these mechanisms could lead to more effective treatments for addictive disorders. PMID- 14746513 TI - Beta-cell replacement for type I diabetes. AB - The ability to achieve insulin independence with either solid-organ pancreas or islet transplantation has increased the number of patients seeking beta-cell replacement as an alternative to insulin therapy. Despite dramatic improvements in the ability to achieve insulin independence following solid-organ pancreas transplantation, the secondary complications of long-standing diabetes are frequently irreversible by the time surgical intervention is justified based on the risk of this procedure. Pancreatic islet transplantation provides a safer and less invasive alternative for beta-cell replacement that could be justified earlier in the course of diabetes to prevent the development of secondary complications. Recent advances in the technology of islet isolation, as well as the ability to prevent the alloimmune and recurrent autoimmune response following islet transplantation with immunosuppressive regimens that are not toxic to beta cells, have rekindled an interest in this field. Widespread application of islet transplantation will depend on further improvements in selective immunosuppression, development of immunologic tolerance, and finding new sources of beta cells. PMID- 14746514 TI - Cochlear implantation for the treatment of deafness. AB - Cochlear implants have dramatically changed the treatment and prognosis for patients with profound sensorineural hearing loss. Deaf adults and children can be successfully (re)integrated into the hearing world through a multidisciplinary approach involving otolaryngologists, audiologists, and speech/language pathologists. As the technology of the cochlear prosthesis advances, the candidacy for these devices continues to broaden. This review addresses the basic technology, candidacy criteria, and important issues in the fields of adult and pediatric cochlear implantation. Cost utility and future directions in the treatment of the profoundly hearing impaired are discussed. PMID- 14746515 TI - Drug-eluting stents. AB - Advances in catheter and stent design have made stent implantation the standard coronary angioplasty procedure. Unfortunately, in-stent restenosis continues to plague this procedure, with the optimum binary restenosis rates reaching ~10% to 20%. In the past few years, it has become clear that in-stent restenosis is largely due to the migration and proliferation of vascular smooth muscle cells to form a neointima. To address this issue, stents coated with drug-delivery vehicles have been developed to deliver antiproliferative therapeutics. Two drugs, rapamycin and taxol, have been the lead compounds for testing the idea of a drug-eluting stent. These drugs have been successful largely because of the solid mechanistic understanding of their effects and extensive preclinical examination. The result of these years of work is that the rapamycin-coated stent entered the US market in April of 2003, and the taxol-coated stent appears poised to follow soon. PMID- 14746516 TI - New approaches to hemodialysis. AB - Treatment of end-stage renal disease with dialysis is characterized by high mortality rate, low quality of life, and high cost. Recent randomized controlled studies showed that increasing the dialysis dose above the currently recommended levels in thrice-weekly hemodialysis does not decrease the patient mortality rate. Short daily hemodialysis or daily home nocturnal hemodialysis are promising alternatives. Both improve quality of life and control blood pressure and anemia; nocturnal hemodialysis additionally controls serum phosphates without phosphate binders, allows a free diet, and corrects sleep apnea. Although the direct cost of daily hemodialysis is higher than that of conventional hemodialysis, the cost of total care, especially when delivered at home, seems to be lower. Further confirmation of these results is important. Restructuring of the dialysis reimbursement system is necessary to make the use of daily hemodialysis possible. Hemofiltration techniques, sorbents, and the renal tubular assist device may also help change the current grim statistics. PMID- 14746517 TI - Emerging infectious threats to the blood supply. AB - During the past 15 years, it has become clear that new agents and new strains of existing agents continue to emerge worldwide as protagonists of infectious disease. These emerging agents pose threats not only to the general human population but also to recipients of blood transfusions. Indeed, the modern era of blood safety perhaps began with the recognition of HIV as an emerging agent transmissible by blood transfusion. Today, emerging infectious agents that pose a threat to the blood supply are not limited to viruses, but include bacterial, protozoan, and prion agents. Preventing the transmission of these new agents by blood transfusion is often problematic, as the available tools may be inadequate. It is certain, however, that new agents will continue to emerge as threats to blood safety and these agents are likely to require novel approaches to prevent their transmission. PMID- 14746518 TI - Lead poisoning. AB - Understanding of lead toxicity has advanced substantially over the past three decades, and focus has shifted from high-dose effects in clinically symptomatic individuals to the consequences of exposure at lower doses that cause no symptoms, particularly in children and fetuses. The availability of more sensitive analytic methods has made it possible to measure lead at much lower concentrations. This advance, along with more refined epidemiological techniques and better outcome measures, has lowered the least observable effect level until it approaches zero. As a consequence, the segment of the population who are diagnosed with exposure to toxic levels has expanded. At the same time, environmental efforts, most importantly the removal of lead from gasoline, have dramatically reduced the amount of lead in the biosphere. The remaining major source of lead is older housing stock. Although the cost of lead paint abatement is measured in billions of dollars, the monetized benefits of such a Herculean task have been shown to far outweigh the costs. PMID- 14746519 TI - The impact of minimally invasive surgical techniques. AB - The use of minimally invasive techniques (MIT) in patient care is well documented in ancient history; however, it was not until the 1990s that advancements in technology enabled surgeons to realize the true potential of this approach. The minimally invasive approach has revolutionized surgical care, significantly reducing postoperative pain, recovery time, and hospital stays with marked improvements in cosmetic outcome and overall cost-effectiveness. It is now used around the world and in all major fields of surgery, compelling changes in training programs in order to assure quality control and patient safety. The bond between surgeons practicing minimally invasive surgery (MIS) and the high-tech industry is of utmost importance to future developments. Surgical robotic systems represent the most technologically advanced product of this collaboration, and their potential application in MIS shows much promise. As technology advances, additional developments in MIT are likely. PMID- 14746520 TI - Implementing a research agenda for complementary and alternative medicine. AB - Complementary and alternative medicine (CAM) consists of diverse clinical interventions that are practiced because of their popularity rather than the prior demonstration of safety and efficacy required for conventional agents. CAM therapies can be grouped into five categories: biologically based therapies, manipulative and body-based interventions, mind-body interventions, "energy" therapies, and alternative medical systems. The present evidence that individual CAM interventions are efficacious is largely anecdotal, but hundreds of small trials have yielded positive results. For a few modalities, existing data are either very encouraging or else sufficient to conclude that they are ineffective. CAM interventions are presumed to be safe, yet they may not be, particularly in the case of botanical agents with inherent toxicities, significant drug interactions, or potent adulterants. The public health questions regarding CAM can only be addressed through a research agenda that defines which interventions have favorable therapeutic indices. Implementation of this agenda involves adequate characterization and standardization of the product or practice, with rigorous investigation to demonstrate its safety, mechanism of action, and efficacy. PMID- 14746522 TI - Clinical management of tuberculosis in the context of HIV infection. AB - Globally, the HIV and tuberculosis epidemics are stoking each other, creating a public health crisis of enormous proportions. At the level of individuals, contemporaneous infection with M. tuberculosis and HIV poses great challenges to clinical management. This chapter provides an overview of active and latent tuberculosis treatment in HIV-infected and -uninfected individuals. The discussion focuses on medication issues, including interactions between antitubercular drugs, antiretroviral drugs, and medicines used for opportunistic infections and treatment in the face of comorbidities. Clinical questions specific to coinfection are discussed, including duration and timing initiation of therapy and immune reconstitution. Most of the data presented were generated in industrialized settings and are presented to assist patient management in such settings. However, given the disproportionate amount of TB/HIV in less-developed nations and the increasing availability of antiretroviral therapy in resource limited settings, the issues presented will become increasingly relevant globally. PMID- 14746523 TI - HIV-associated lipodystrophy: pathogenesis, prognosis, treatment, and controversies. AB - Potent antiretroviral agents markedly suppress HIV and have dramatically improved the clinical course, prognosis, and survival of HIV-infected patients. Unfortunately, highly active antiretroviral therapy is often compromised by metabolic complications, including insulin resistance, dyslipidemia, and fat redistribution. Together these changes have been termed the HIV-lipodystrophy syndrome, which is estimated to affect a majority of patients treated with potent combination antiretroviral therapy. Routine testing of fasting glucose is recommended for all HIV-infected patients, particularly those who are obese, have a family history of diabetes mellitus, or are receiving protease inhibitor therapy. Preliminary investigations have demonstrated the potential utility of insulin-sensitizing agents and lipid-lowering therapies to ameliorate these metabolic disturbances. Patients with HIV infection who demonstrate fat redistribution and develop hyperinsulinemia and dyslipidemia may be at increased risk of cardiovascular disease. However, the long-term effects on cardiovascular disease have not yet been determined. PMID- 14746521 TI - Basic advances and new avenues in therapy of spinal cord injury. AB - The prospects for successful clinical trials of neuroprotective and neurorestorative interventions for patients with acute and chronic myelopathies depend on preclinical animal models of injury and repair that reflect the human condition. Remarkable progress continues in the attempt to promote connections between the brain and the sensory and motor neurons below a spinal cord lesion. Recent experiments demonstrate the potential for biological therapies to regenerate or remyelinate axons and to incorporate new neural cells into the milieu of a traumatic spinal cord injury. The computational flexibility and plasticity of the sensorimotor systems of the brain, spinal cord, and motor unit make functional use of new circuitry feasible in patients. To incorporate residual and new pathways, neural repair strategies must be coupled to rehabilitation therapies that drive activity-dependent plasticity for walking, for reaching and grasping, and for bowel and bladder control. Prevention of pain and dysautonomia are also clinical targets. Research aims to define the temporal windows of opportunity for interventions, test the safety and efficacy of delivery systems of agents and cells, and provide a better understanding of the cascades of gene expression and cell interactions both acutely and chronically after injury. These bench-to-bedside studies are defining the neurobiology of spinal cord injury rehabilitation. PMID- 14746524 TI - Human papillomavirus vaccines and prevention of cervical cancer. AB - Cervical cancer and precancerous cervical lesions constitute a major problem in women's health. Every year 470,000 cases of cervical cancer are diagnosed worldwide, and about half the women afflicted will die. In the United States alone, approximately 14,000 cases of cervical cancer are diagnosed each year despite the availability of screening and access to high-quality gynecological care. With the confirmation that cervical cancer is caused by an infectious agent, human papillomavirus, the possibility of fighting this disease with either prophylactic or therapeutic vaccination arose. This review describes advances in vaccine development and very promising first results for prophylactic vaccination against cervical cancer. PMID- 14746525 TI - Opportunities for control of meningococcal disease in the United States. AB - The United States currently has relatively low rates of meningococcal disease caused by Neisseria meningitidis. Serogroups Y, C, and B are most common. Although most cases are sporadic, a minority are associated with outbreaks. Pediatric populations have disproportionately higher rates of disease, but nearly two thirds of all cases occur in persons aged 15 years and older. The major challenge to control of domestic meningococcal disease is the absence of a vaccine to prevent sporadic cases spanning many age groups. The quadrivalent A/C/Y/W-135 meningococcal polysaccharide vaccine is licensed in the United States, but because of its limited efficacy in children under two years of age, it is recommended for high-risk groups and outbreak response rather than routine childhood immunization. New conjugate meningococcal vaccines have successfully reduced endemic disease in the United Kingdom, and similar vaccines promise to have a dramatic impact on the burden of meningococcal disease in the United States. PMID- 14746526 TI - Recent advances in the development of HIV-1 vaccines using replication incompetent adenovirus vectors. AB - An increasing body of evidence suggests that a vaccine that elicits anti-HIV-1 cellular immunity could provide the basis for an effective AIDS vaccine. Comparative immunization experiments testing a variety of vaccine approaches have demonstrated that replication-incompetent adenovirus vectors are an effective means for eliciting cytotoxic T-lymphocyte (CTL) immune responses against HIV-1 antigens. These immune responses effectively control viremia in nonhuman primates following challenge with simian AIDS viruses. Such data, coupled with epidemiology studies that identify HIV-1 gag, pol, and nef as the best antigens for broadly directed cellular immune responses, provide guidance for the development of a potential AIDS vaccine. PMID- 14746527 TI - Left ventricular diastolic dysfunction and diastolic heart failure. AB - Thirty to fifty percent of patients presenting with signs and symptoms of heart failure have a normal left ventricular (LV) systolic ejection fraction. The clinical examination cannot distinguish these patients (diastolic heart failure) from those with a depressed ejection fraction (systolic heart failure), but echocardiography can. The management of diastolic heart failure has two major objectives. The first is to reverse the consequences of diastolic dysfunction (e.g., venous congestion), and the second is to eliminate or reduce the factors responsible for diastolic dysfunction (e.g., myocardial hypertrophy, fibrosis, and ischemia). PMID- 14746528 TI - Mechanisms of pulmonary fibrosis. AB - Tissue injury evokes highly conserved, tightly regulated inflammatory responses and less well-understood host repair responses. Both inflammation and repair involve the recruitment, activation, apoptosis, and eventual clearance of key effector cells. In this review, we propose the concept of pulmonary fibrosis as a dysregulated repair process that is perpetually "turned on" even though classical inflammatory pathways may be dampened or "switched off." Significant regional heterogeneity, with varied histopathological patterns of inflammation and fibrosis, has been observed in individual patients with idiopathic pulmonary fibrosis. We discuss environmental factors and host response factors, such as genetic susceptibility and age, that may influence these varied manifestations. Better understanding of the mechanisms of lung repair, which include alveolar reepithelialization, myofibroblast differentiation/activation, and apoptosis, should offer more effective therapeutic options for progressive pulmonary fibrosis. PMID- 14746529 TI - Systemic mastocytosis. AB - Systemic mastocytosis is a clonal disorder of the mast cell and its progenitor. The symptoms of systemic mastocytosis are due to the pathologic accumulation and activation of mast cells in various tissues such as bone marrow, skin, gastrointestinal tract, liver, and spleen. Recent studies revealed striking differences between the molecular and cellular biology of mast cells in patients with mastocytosis and those of healthy individuals. These findings are being used in formulating diagnostic criteria as well as designing novel treatment approaches to the disease. PMID- 14746530 TI - The erbB family: targets for therapeutic development against cancer and therapeutic strategies using monoclonal antibodies and tyrosine kinase inhibitors. AB - The overexpression and aberrant function of members of the erbB family of receptors, particularly erbB1 (also known as epidermal growth factor receptor), and its ligands in many human cancers have provided a rationale for targeting this signaling network with novel approaches. erbB1 is a selective target for inhibiting cancers because its activation often confers a proliferative advantage. Activation of the erbB1 tyrosine kinase provides signals that drive dysregulated proliferation, invasion, metastasis, angiogenesis, and cell survival, and its inhibition has potential in both the treatment and prevention of these malignancies. Based on the structure and function of erbB1, two therapeutic strategies have been developed. The first uses human monoclonal antibodies (MAbs) generated against the receptor's ligand-binding extracellular domain. These MAbs block binding of receptor-activating ligands, and, in some cases, can induce receptor endocytosis and downregulation. The second uses small molecules that compete with adenosine triphosphate (ATP) for binding to the receptor's kinase pocket, thereby blocking receptor activation and the transduction of postreceptor signals. Early clinical studies suggest that both approaches are well tolerated and can induce clinical activity in many common malignancies. PMID- 14746531 TI - Nonmyeloablative allogeneic immunotherapy for solid tumors. AB - Over the past decade, considerable advances have been made in the field of allogeneic hematopoietic stem cell transplantation. Recognition that transplanted donor immune cells can cure patients with leukemia has led to the development of nonmyeloablative or "low-intensity" conditioning regimens, which have expanded the application of allogeneic transplantation to a growing number of hematological malignancies. The improved safety and preliminary success of this transplant approach have justified applying allogeneic immunotherapy to patients with treatment-refractory solid tumors. PMID- 14746532 TI - Rituximab: expanding role in therapy for lymphomas and autoimmune diseases. AB - Rituximab (Rituxan) is a human-mouse chimeric monoclonal antibody that targets the B-cell CD20 antigen and causes rapid and specific B-cell depletion. Rituximab was approved in the United States in 1997 to treat low-grade or follicular, relapsed or refractory, CD20-positive B-cell non-Hodgkin's lymphoma (NHL). Since then, further clinical experience with rituximab has been incorporated into the prescribing information, which now stipulates an extended eight-week schedule, treatment of patients with refractory or relapsed bulky disease measuring >10 cm, and retreatment of patients who responded to rituximab previously. In 1998, the European Union approved rituximab (MabThera) to treat stage III/IV, follicular, chemotherapy-resistant, or relapsed NHL. Recently, the European Union also approved the use of rituximab in combination with standard chemotherapy for aggressive NHL. Many clinical trials have evaluated rituximab, alone or with other therapies, in indolent and aggressive NHL as well as other B-cell lymphoproliferative disorders. New studies are evaluating rituximab's role in first-line therapy, maintenance therapy, and stem-cell transplantation procedures. The use of rituximab against autoimmune disorders, such as rheumatoid arthritis, immune thrombocytopenic purpura, autoimmune hemolytic anemia, systemic lupus erythematosus, and multiple sclerosis, is also under investigation. PMID- 14746533 TI - Botulinum toxin and other new approaches to migraine therapy. AB - The number of migraine treatments and our understanding of migraine pathophysiology are both increasing. Newer treatments focus on migraine prevention. Botulinum toxin (BTX) is a potent neurotoxin used primarily to treat diseases associated with increased muscle activity. Recently, BTX was found to have antinociceptive effects that are probably independent of its muscle-relaxant action. Clinical trials support the efficacy of BTX type A (and possibly also type B) in the treatment of migraine. The anticonvulsant topiramate was recently shown to be effective for migraine prevention. At the low doses used for this indication, cognitive side effects are not a major concern. Another new approach to migraine prevention is angiotensin type 1 (AT1) receptor blockade. The high tolerability of the AT1 receptor blocker candesartan warrants further studies to assess its role in migraine prevention. PMID- 14746534 TI - Management of infections in the neutropenic patient. AB - Neutropenic patients continue to be at increased risk for developing serious infections despite substantial advances in supportive care. Epidemiologic shifts occur periodically and need to be detected early because they influence prophylactic, empiric, and specific therapy strategies. Although effective in preventing bacterial and some fungal infections, prophylaxis must be used with caution because it is associated with the emergence of resistance. The choices for empiric therapy include combination regimens and monotherapy. Specific choices depend on local factors (epidemiology, susceptibility/resistance patterns, availability). Various treatment settings (hospital-based, early discharge, outpatient) are also available, and the choice depends on the patient's risk category. Early diagnosis and treatment of many fungal and viral infections remains suboptimal. Infection control and prevention are important strategies, especially with the emergence of multidrug-resistant organisms. PMID- 14746535 TI - Diagnostic PCR: making internal amplification control mandatory. PMID- 14746536 TI - Regulation of pyrimidine nucleotide formation in Pseudomonas reptilivora. AB - AIMS: To study the regulation of de novo pyrimidine biosynthesis in the pathogenic bacterium Pseudomonas reptilivora ATCC 14836. METHODS AND RESULTS: The pyrimidine biosynthetic pathway enzymes were assayed in extracts of Ps. reptilivora ATCC 14836 cells and of cells from an auxotroph lacking aspartate transcarbamoylase activity. Pyrimidine biosynthetic pathway enzyme activities in ATCC 14836 were influenced by the addition of pyrimidine bases to the culture medium with orotic acid addition inducing dihydroorotase activity. Pyrimidine starvation of the transcarbamoylase mutant strain increased its de novo enzyme activities suggesting that the de novo pathway was also subject to repression by a pyrimidine-related compound. Aspartate transcarbamoylase activity in ATCC 14836 was inhibited in vitro by pyrophosphate and ATP. CONCLUSIONS: Regulation of pyrimidine biosynthesis in Ps. reptilivora was observed at the level of enzyme synthesis and at the level of activity for aspartate transcarbamoylase. Its regulation of enzyme synthesis seemed to be more highly controlled than what was observed in the related species Ps. fluorescens. SIGNIFICANCE AND IMPACT OF THE STUDY: This investigation found that pyrimidine biosynthesis is controlled in Ps. reptilivora. This could prove helpful to future studies exploring its pathogenicity. PMID- 14746537 TI - Chemical composition and microbicidal activity of extracts from Brazilian and Bulgarian propolis. AB - AIMS: The chemical composition of ethanol extracts from a Brazilian (Et-Bra) and a Bulgarian (Et-Blg) propolis, and their activity against the protozoan Trypanosoma cruzi, several fungi and bacteria species were determined. METHODS AND RESULTS: The chemical composition was determined by high temperature high resolution gas chromatography coupled to mass spectrometry. Microbiological activity was assayed in vitro against T. cruzi, Candida albicans, Sporothrix schenckii, Paracoccidioides brasiliensis, Neisseria meningitidis, Streptococcus pneumoniae and Staphylococcus aureus. CONCLUSIONS: Et-Bra and Et-Blg, although with totally distinct compositions, were active against T. cruzi and the three species of fungi. Et-Blg was more effective than Et-Bra against bacteria, particularly N. meningitidis and Strep. pneumoniae. SIGNIFICANCE AND IMPACT OF THE STUDY: Although with different classes of components, both propolis extracts showed microbicidal activity. For the bactericidal activity it was possible to establish a positive correlation with the high content of flavonoids of the Bulgarian extract. PMID- 14746538 TI - Influence of petroleum contamination and biostimulation treatment on the diversity of Pseudomonas spp. in soil microcosms as evaluated by 16S rRNA based PCR and DGGE. AB - AIMS: The aim of this study was to apply a group specific PCR system followed by denaturing gradient gel electrophoresis (DGGE) analysis to evaluate the effect of oil contamination and the biostimulation process on the diversity of Pseudomonas populations in soil ecosystems. METHODS AND RESULTS: Direct DNA extraction from biostimulated- and oil-contaminated soil samples was performed. Primers specific for the genus Pseudomonas spp. were used to amplify 16S rRNA genes and then a semi-nested PCR reaction was applied to obtain smaller fragments for comparing the PCR products by DGGE. Whether in bulk, oil-contaminated or biostimulated soils, the DGGE profiles revealed little change in Pseudomonas community throughout the 270 days of experiment. The presence of a few additional bands observed only in treated samples indicated that a bacterial shift occurred with the addition of nutrients and with oil contamination. CONCLUSIONS, SIGNIFICANCE AND IMPACT OF THE STUDY: The combination of semi-nested PCR and DGGE was found to be a rapid and sensitive technique to study the diversity within the genus Pseudomonas and may be suitable for further studies concerning the role of this bacterial group in large-scale oil-contaminated areas. PMID- 14746539 TI - Study of two bacteriocins produced by Enterococcus casseliflavus and Ent. faecalis. AB - AIMS: The antimicrobial activity of two plasmid-borne bacteriocins produced by Enterococcus casseliflavus IM 416K1 and Ent. faecalis IM 388C and their mating transferability were studied. METHODS AND RESULTS: Both bacteriocins showed antibacterial activity against taxonomically related micro-organisms and Listeria monocytogenes but differ for heat sensitivity, antimicrobial titre, molecular size and class of affiliation. The transferability by mating of the antibacterial properties from producers to Enterococcus faecalis JH2-2 revealed that the bacteriocin-phenotype was linked in both strains to genes located on a 34 MDa plasmid. This result was confirmed by loss of antibacterial activity and immunity after curing treatment. CONCLUSIONS: Restriction analysis has shown a different profile of the two conjugative plasmids. Enterocin 416K1 and Enterocin 388C could represent natural antilisterial agents to use in food technology. SIGNIFICANCE AND IMPACT OF THE STUDY: The transferability of the 34 MDa conjugative plasmids might be considered a possibility for the study of bacteriocins expression in bacterial hosts different from the native strains. PMID- 14746540 TI - Growth kinetics of aerobic granules developed in sequencing batch reactors. AB - AIMS: This paper attempts to develop a kinetic model to describe the growth of aerobic granules developed under different operation conditions. METHODS AND RESULTS: A series of experiments were conducted by using four-column sequencing batch reactors to study the formation of aerobic granules under different conditions, e.g. organic loading rates, hydrodynamic shear forces and substrate N/COD ratios. A simple kinetic model based on the Linear Phenomenological Equation was successfully derived to describe the growth of aerobic granules. It was found that the growth of aerobic granules in terms of equilibrium size and size-dependent growth rate were inversely related to shear force imposed to microbial community, while a high organic loading favoured the growth of aerobic granules, leading to a large size granule. The effect of substrate N/COD ratio on the growth kinetics of aerobic granules was realized through change in microbial populations, and enriched nitrifying population in aerobic granules developed at high substrate N/COD ratio resulted in a low overall growth rate of aerobic granules. CONCLUSIONS: The proposed model can provide good prediction for the growth of aerobic granules indicated by the correlation coefficient >0.95. SIGNIFICANCE AND IMPACT OF THE STUDY: The kinetic model proposed could offer a useful tool for studying the growth kinetics of cell-to-cell immobilization process. The study confirmed that the growth of aerobic granules and biofilms are subject to a similar kinetic pattern. This work would also be helpful for better understanding the mechanism of aerobic granulation. PMID- 14746541 TI - Occurrence and dominance of yeast species in sourdough. AB - AIMS: The aim of this work is to identify the dominant yeast species in homemade sourdoughs. METHODS AND RESULTS: PCR/restriction fragment length polymorphism analysis of internal transcribed spacer regions was used for the identification of isolates and the data were confirmed with phenotypic tests. The strains belonging to Saccharomyces cerevisiae were identified to strain level by analysis of inter-delta regions. CONCLUSION: This work shows that the dominant species in homemade sourdoughs can differ from each other. Saccharomyces cerevisiae was found to be the dominant species, followed by the Candida milleri, C. humilis, S. exiguus and Issatchenkia orientalis. The inter-delta regions of S. cerevisiae strains showed high polymorphism. SIGNIFICANCE AND IMPACT OF THE STUDY: Occurrence of single, non-Saccharomyces species and S. cerevisiae polymorphism in the yeast populations of sourdough samples. PMID- 14746542 TI - Rapid detection of Oenococcus oeni in wine by real-time quantitative PCR. AB - AIMS: To develop a real-time polymerase chain reaction (PCR) method for rapid detection and quantification of Oenococcus oeni in wine samples for monitoring malolactic fermentation. METHODS AND RESULTS: Specific primers and fluorogenic probe targeted to the gene encoding the malolactic enzyme of O. oeni were developed and used in real-time PCR assays in order to quantify genomic DNA either from bacterial pure cultures or wine samples. Conventional CFU countings were also performed. The PCR assay confirmed to be specific for O. oeni species and significantly correlated to the conventional plating method both in pure cultures and wine samples (r = 0.902 and 0.96, respectively). CONCLUSIONS: The DNA extraction from wine and the real-time PCR quantification assay, being performed in ca 6 h and allowing several samples to be concurrently processed, provide useful tools for the rapid and direct detection of O. oeni in wine without the necessity for sample plating. SIGNIFICANCE AND IMPACT OF THE STUDY: Rapid quantification of O. oeni by a real-time PCR assay can improve the control of malolactic fermentation in wines allowing prompt corrective measures to regulate the bacterial growth. PMID- 14746543 TI - Species-specific PCR method for identification of Streptococcus downei. AB - AIMS: To establish a rapid method to differentiate Streptococcus downei and S. sobrinus by multiplex PCR. METHODS AND RESULTS: A PCR primer pair specific to S. downei was designed on the basis of the nucleotide sequence of the dextranase gene of S. downei NCTC 11391T. The primer pair specifically detected S. downei, but none of the other mutans streptococci (16 strains of six species). The PCR procedure was capable of detecting 1 pg of genomic DNA purified from S. downei NCTC 11391 and as few as 14 CFU of S. downei cells. The mixture of primer pairs specific to each S. downei (this study) and S. sobrinus (Igarashi et al. 2000) detected only the strains of these two species among all the mutans streptococcal strains, and concomitantly differentiated the two species by species-specific amplicons of different lengths. CONCLUSIONS: The present PCR method is highly specific to S. downei and is useful for detection and identification of S. downei. SIGNIFICANCE AND IMPACT OF THE STUDY: Multiplex PCR using dextranase gene primers is a useful method for simultaneous detection and differentiation of S. downei and S. sobrinus. PMID- 14746544 TI - rpoB-PCR amplified gene and temporal temperature gradient gel electrophoresis: a rapid tool to analyse bacterial strains representative of cold-smoked salmon microflora. AB - AIM: To evaluate rpoB gene as a biomarker of microbial biodiversity associated to cold-smoked salmon by a novel nested-polymerase chain reaction/temporal temperature gradient gel electrophoresis (PCR/TTGE) technique applied on pure cultures of reference strains. METHODS AND RESULTS: DNA obtained from pure cultures of reference strains was used in a succession of a first PCR amplification of rpoB fragment with degenerated nonclamped primers and a nested PCR with nondegenerated clamped primers. PCR products were then applied on a TTGE gel in order to analyse strains profile. High quantity of nested-PCR products were obtained for each tested strain and TTGE profiles showed a good separation between the different reference bacteria and an easy way to associate one band to one species. CONCLUSION: The nested-PCR/TTGE technique used in this study is a promising way of investigating bacterial community structure of cold-smoked salmon or other food matrix. SIGNIFICANCE AND IMPACT OF THE STUDY: Because of its single copy state leading to single band profiles in TTGE, rpoB constitute a good potential molecular marker for further development of cold-smoked salmon biodiversity analysis. PMID- 14746545 TI - Thermal characteristics of recombinant green fluorescent protein (GFPuv) extracted from Escherichia coli. AB - AIMS: The thermal stability of isolated and extracted recombinant green fluorescent protein (GFPuv) was evaluated by analysing the loss of fluorescence intensity. METHODS AND RESULTS: GFPuv was expressed by Escherichia coli, extracted by the three-phase partitioning method and purified by elution through an hydrophobic interaction column. The collected fractions were further diluted in Tris-HCl-EDTA (pH 8.0) and subjected to continuous heating at set temperatures (45-95 degrees C). From a standard curve relating fluorescence intensity to GFPuv concentration, the loss of fluorescence intensity was converted to denatured GFPuv concentration (microg ml-1). To determine the extent of the thermal stability of GFPuv, decimal reduction times (D-values), z-value and energy of activation (Ea) were calculated. CONCLUSIONS: For temperatures between 45 and 70 degrees C, extracted native GFPuv activity decreased from 11 to 75% relative to initial native protein concentration above 70 degrees C, the average decrease in GFPuv fluorescence was between 72 to 83%. SIGNIFICANCE AND IMPACT OF THE STUDY: The thermal stability of GFPuv provides the basis for its potential utility as a fluorescent biological indicator to assess the efficacy of the treatment of liquids and materials exposed to steam. PMID- 14746547 TI - Identification of heat stable protease of Klebsiella oxytoca isolated from raw milk. AB - AIMS: The aim of this study was to identify and characterize heat stable proteinases of psychrotrophic proteolytic bacteria isolated from raw milk. METHODS AND RESULTS: A strain of Klebsiella oxytoca producing a high proteolytic activity when cultured on milk was isolated. Maximum proteolytic activity was observed at the stationary phase during growth on milk or casein-peptone broth. The bacterium demonstrated the capability to grow at 7 degrees C, classified as psychrotrophic. The crude enzyme showed optimum activity at 37 degrees C, and pH 5.0 and 7.0. The proteinase was very resistant to heat, maintaining 74% of initial activity after incubation at 142 degrees C. CONCLUSIONS: A heat stable protease of a psychrotrophic strain of K. oxytoca was identified and partially characterized. SIGNIFICANCE AND IMPACT OF THE STUDY: Thermal stable proteases may constitute a serious problem to ultra-high temperature (UHT) processed milk, leading to undesirable physical and sensory alterations. PMID- 14746546 TI - Characterization of antimicrobial resistant Salmonella Kinshasa from dairy calves in Texas. AB - AIM: To determine the prevalence of antimicrobial resistance among Salmonella isolated from a central Texas dairy calf farm that raises animals for dairy-beef production. METHODS AND RESULTS: Salmonella isolates collected from 50 faecal samples were characterized for susceptibility to 20 antimicrobial agents. Seventy per cent of the faecal samples (35 of 50) tested positive for Salmonella, and high rates of resistance to the following drugs that are commonly used for treatment of bacterial enteritis in livestock were observed: ampicillin (88%), apramycin (83%), neomycin (86%), spectinomycin (91%) and oxytetracycline (90%). No resistance to the fluoroquinolone antibiotics was observed. The most prevalent Salmonella serotype was Kinshasha (22 of 35 samples), followed by Agona (4 of 35), Newport (3 of 35), Infantis (2 of 35), Montevideo (2 of 35), Lille (1 of 35) and Newington (1 of 35). The Kinshasa, Agona, Newport and Infantis serotypes all displayed resistance to ampicillin, chloramphenicol, streptomycin, sulphamethoxazole and tetracycline, and the penta-resistance phenotype was transferable to an Escherichia coli recipient strain. SIGNIFICANCE: Multi-drug resistant Salmonella in dairy calves pose a costly animal health problem and a potential risk to the public health. This study emphasizes the need for alternative, non-antimicrobial intervention strategies for the control of zoonotic pathogens. PMID- 14746548 TI - Identification of a gene encoding a putative phosphotransferase system enzyme IIBC in Listeria welshimeri and its application for diagnostic PCR. AB - AIMS: To identify a Listeria welshimeri-specific gene that can be used for identification of this species by PCR. METHODS AND RESULTS: Through comparative analysis of genomic DNA from Listeria species using dot blot hybridization, an L. welshimeri-specific clone was isolated that contained a gene segment whose translated protein sequence is similar to enzyme IIBC from phosphotransferase systems in other bacteria. Using oligonucleotide primers derived from this L. welshimeri-specific clone, a 608-bp fragment was amplified from L. welshimeri genomic DNA and not from other Listeria species or other Gram-negative and Gram positive species. CONCLUSION AND SIGNIFICANCE: The PCR employing L. welshimeri specific primers shows promise as a useful method for differentiating L. welshimeri from other Listeria species and related bacteria. PMID- 14746549 TI - The effect of hydraulic retention time on the stability of aerobically grown microbial granules. AB - AIMS: The aim of this study is to evaluate the effect of hydraulic retention time (HRT) on the development of aerobically grown microbial granules. METHODS AND RESULTS: Five column-shaped sequential aerobic sludge blanket reactors (SASBRs) were seeded with aerobically grown microbial granules and operated in a cyclic mode at different HRTs. At the shortest HRT of 1 h, the strong hydraulic pressure triggered biomass washout and led to reactor failure. At the longest HRT of 24 h, which represented the weakest hydraulic selection in this study, aerobic granules were gradually substituted by bioflocs because of the lower frequency of volumetric exchange. Within the optimum range of HRTs from 2 to 12 h, however, aerobic granules became stabilized in the presence of adequate hydraulic selection in the reactors, with good mixed liquor volatile suspended solids (MLVSS) retention, high volumetric chemical oxygen demand (COD) removal, low sludge volume index (SVI) values, good effluent quality, low sludge production rate, stronger and more compact structures, high cell hydrophobicity and high ratios of extracellular polysaccharides (PS) to extracellular proteins (PN). CONCLUSIONS: HRTs between 2 and 12 h provided the hydraulic selection pressures favourable for the formation and maintenance of stable aerobic granules with good settleability and activity. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first systematic study on the effect of HRT on heterotrophic aerobic granules. The results of the investigation are useful in understanding how aerobic granules can be applied for wastewater treatment. PMID- 14746550 TI - A cryopreservation procedure for the rumen protozoon Entodinium caudatum: estimation of its viability by fluorescence microscopy. AB - AIMS: To study the viability of a culture of the rumen protozoon Entodinium caudatum after a cryopreservation procedure by a fluorescence microscopy staining method. METHODS AND RESULTS: Fluorescence method is based on the different colour of cells depending on their membrane integrity. When the temperature effect was studied either by fluorescence or motility, the techniques were correlated (r = 0.727) and their slopes and intercepts were not different (P > 0.05). However, motility showed a higher variation coefficient (0.40 vs 0.12). There were no differences between cooling rates at cryopreservation (1 and 4 degrees C min-1) at 38, 15 or 5 degrees C, nor after thawing. CONCLUSIONS: Fluorescence staining is more accurate than motility for assessing protozoal viability. Viability after thawing was 0.50, and the number of viable cells per 250 microl straw was 320 and 420 for 1 and 4 degrees C min-1. SIGNIFICANCE AND IMPACT OF THE STUDY: This cryopreservation procedure seems to ensure culture recovery for E. caudatum. PMID- 14746551 TI - Trichoderma harzianum enhances the production of nematicidal compounds in vitro and improves biocontrol of Meloidogyne javanica by Pseudomonas fluorescens in tomato. AB - AIMS: To determine the influence of soil-borne fungus Trichoderma harzianum on the biocontrol performance of Pseudomonas fluorescens strain CHA0 and its 2,4 diacetylphloroglucinol (DAPG) overproducing derivative CHA0/pME3424 against Meloidogyne javanica. METHODS AND RESULTS: Amendment of the culture filtrate (CF) or methanol extract of the CF of a T. harzianum strain Th6 to P. fluorescens growth medium enhanced the production of nematicidal compound(s) by bacterial inoculants in vitro. In addition, bacteria overwhelmingly expressed phl'-'lacZ reporter gene when the medium was amended with CF of T. harzianum. Pseudomonas fluorescens and T. harzianum applied together in unsterilized sandy loam soil caused greater reduction in nematode population densities in tomato roots. CONCLUSIONS: Trichoderma harzianum improves root-knot nematode biocontrol by the antagonistic rhizobacterium P. fluorescens both in vitro and under glasshouse conditions. SIGNIFICANCE AND IMPACT OF THE STUDY: The synergistic effect of T. harzianum on the production of nematicidal compound(s) critical in biocontrol may improve the efficacy of biocontrol bacteria against plant-parasitic nematodes. Considering the inconsistent performance of the biocontrol agents under field conditions, application of a mixture of compatible T. harzianum and P. fluorescens would more closely mimic the natural situation and might broaden the spectrum of biocontrol activity with enhanced efficacy and reliability of control. PMID- 14746552 TI - Novel method for cell immobilization and its application for production of oligosaccharides from sucrose. AB - AIMS: The purpose of the present investigation was to develop a novel method for cell immobilization. METHODS AND RESULTS: Aureobasidium pullulans cells were mixed with an alginate solution, and the mixture was extruded to form small gel beads as hydrated-immobilized cells. The beads were then placed at -15 degrees C for 6-24 h to induce freeze-dehydration. The freeze-dehydration resulted in shrinkage of beads as a result of water removal reducing bead volume by 82% and bead weight by 85%. The dehydrated beads were successfully used for the production of fructo-oligosaccharides in a model reactor system. CONCLUSIONS: Dehydrated beads may provide some commercial advantages over conventional immobilized cells. SIGNIFICANCE AND IMPACT OF THE STUDY: This study shows that bioreactor performance can be improved up to two times by the use of the dehydrated beads. PMID- 14746553 TI - Where are clinical trials going? Society and clinical trials. AB - Clinical trials now increasingly impinge on society at large. First there is growing emphasis from health organizations on the need for unbiased evidence about the effectiveness of promoted remedies. Second, as most novel treatments accrue increased costs to society, these need to be evaluated in terms of value for money. Third, there has been confusion and concern about the resolution of conflicting evidence, especially the role of advertising and commercial pressures from a powerful pharmaceutical industry motivated by profit. Fourth, there is concern about research fraud and the ethics of clinical trials. Fifth, there is increasing suspicion of political advice, which sometimes has sought to reassure an anxious public on the basis of complex and possibly inadequate scientific information. Some of these issues are addressed by truly independent and properly constituted data and safety monitoring committees, which are of particular importance when academic investigators or universities have a large financial conflict of interest. This is now more problematic with the current encouragement of investigator-led spin-off companies. These issues are best resolved by independent financial support (from government or other institutions) rather than relying on the commercial sponsor. PMID- 14746554 TI - The systemic amyloidoses: clearer understanding of the molecular mechanisms offers hope for more effective therapies. AB - Knowledge about the systemic amyloidoses has increased considerably during the last few years. This group of diseases is characterized by great biochemical variability, including at least 11 different amyloid fibril proteins and a remarkable range of clinical manifestations. With the understanding that the pathogenesis is different in the various forms of amyloidosis, it is now being increasingly accepted that an early and accurate diagnosis, including that of the underlying biochemical nature, is crucial for a successful treatment. The elucidation of the molecular mechanisms involved in amyloidogenesis is at the basis of the recent blossoming of new, innovative and more effective therapeutic approaches. PMID- 14746555 TI - Metformin and lactic acidosis: cause or coincidence? A review of case reports. AB - OBJECTIVE: Metformin has been associated with the serious side-effect lactic acidosis. However, it remains unclear whether the use of metformin was a cause or a coincidence in lactic acidosis. DESIGN: A literature search of the Index Medicus (1959-66) and of the databases Embase, Medline, Medline Express (1966-99) was performed using the keywords metformin, biguanides and lactic acidosis. All articles of cases with metformin-induced lactic acidosis (MILA) were cross referenced. SUBJECTS: Cases were included for analysis if they met the following criteria: serum pH < or =7.35, lactate concentration > or =5 mmol L(-1). INTERVENTION: A forum of six experts in intensive care medicine independently categorized the cases in MILA unlikely (score 0), possible MILA (score 1) or probable MILA (score 2). MAIN OUTCOME MEASURES: Statistical analysis included the paired interobserver agreement (kappa) and multivariate regression analysis. RESULTS: Of 80 reported cases, 33 were excluded because of insufficient quality. The forum scores of the remaining 47 cases were distributed normally with a mean score of 7 (range 2-10). The kappa-value was 0.041 (SD = 0.24, range -0.514, 0.427). Neither lactate concentration nor mortality correlated with serum metformin concentrations. CONCLUSIONS: Given the low interobserver agreement and the lack of any relationship between metformin levels and outcome parameters, the concept that there is a simple, causal relationship between metformin use and lactic acidosis in diabetic patients has to be reconsidered. PMID- 14746556 TI - Apolipoprotein B and apolipoprotein A-I: risk indicators of coronary heart disease and targets for lipid-modifying therapy. AB - Although LDL cholesterol (LDL-C) is associated with an increased risk of coronary heart disease, other lipoproteins and their constituents, apolipoproteins, may play an important role in atherosclerosis. Elevated levels of apolipoprotein (apo) B, a constituent of atherogenic lipoproteins, and reduced levels of apo A I, a component of anti-atherogenic HDL, are associated with increased cardiac events. Apo B, apo A-I and the apo B/apo A-I ratio have been reported as better predictors of cardiovascular events than LDL-C and they even retain their predictive power in patients receiving lipid-modifying therapy. Measurement of these apolipoproteins could improve cardiovascular risk prediction. PMID- 14746557 TI - QRS prolongation on surface ECG and brain natriuretic peptide as indicators of left ventricular systolic dysfunction. AB - OBJECTIVE: To determine whether the combination of prolonged QRS duration and brain natriuretic peptide (BNP) levels predicts left ventricular systolic dysfunction (LVSD) with a higher accuracy compared with QRS duration or BNP alone. SETTING: University hospital. METHODS: We studied 128 consecutive patients with suspected cardiac disease. At rest the QRS duration on 12-lead ECG and BNP levels were determined. A left ventricular ejection fraction (LVEF) <50% at echocardiography was defined as LVSD. RESULTS: QRS duration in the LVSD group (n = 66, LVEF 30 +/- 8%) was longer than in the group without LVSD (n = 62, LVEF 60 +/- 5%; QRS 129 +/- 34 vs. 96 +/- 20 ms, P < 0.001). BNP was higher in the LVSD group compared with controls (467 +/- 397 vs. 169 +/- 242 pg mL(-1), P < 0.001). A QRS duration of >0.1, >0.11 or >0.12 s was highly specific (63, 90 and 98%) but less sensitive (84, 81 and 75%) for the prediction of LVSD. A QRS cut-off value of 106 ms was moderately sensitive (65%) but very specific (87%) for the prediction of LVSD, whereas a BNP cut-off value of >84 pg mL(-1) was highly sensitive (89%) but only modestly specific (58%). The positive likelihood ratio for LVSD of abnormal BNP (2.0) and QRS prolongation >0.1 s (2.3) was improved by the combination of both criteria (5.1). In multivariate analysis, BNP and QRS duration were independent predictors of LVSD. CONCLUSIONS: The combination of abnormal BNP and QRS prolongation yields a higher positive likelihood ratio for the detection of LVSD compared with the two criteria alone. PMID- 14746558 TI - Sudden cardiac death in the young in Sweden: electrocardiogram in relation to forensic diagnosis. AB - OBJECTIVES: To study electrocardiogram (ECG) in relation to forensic diagnosis in young persons who suffered a sudden cardiac death (SCD) in Sweden during 1992-99. DESIGN: A register study of a national database of forensic medicine in Sweden, selecting all cases of SCD 15-35 years of age. In this group, 12-lead ECGs and clinical data were searched for in military conscription and medical records. The ECGs were re-analysed and classified according to the Minnesota code criteria. SETTING: The whole nation of Sweden. SUBJECTS: Sudden cardiac death victims (66 individuals), 15-35 years of age, where it was possible to obtain an ECG recording. RESULTS: We observed major or minor ECG abnormalities in 82% of the subjects. The most common changes were T wave abnormalities (35%), ST segment changes (32%) and conduction defects (20%). The ECGs were evaluated as pathological in 50% of the cases, more often in arrhythmogenic right ventricular cardiomyopathy (88%) and hypertrophic cardiomyopathy (82%). Cardiac-related symptoms were seen in 76% of the total group and there was a family history of a similar cardiac condition in 18%. CONCLUSIONS: Pathological ECGs were common in young SCD victims, in spite of being taken many years before death. An ECG could help identify prospective victims of SCD, and should always be taken in cases with possible cardiac-related symptoms or a family history of SCD. The pathological ECGs were often found in connection with routine screening at military enlistment for men, which raises the question of a routine screening in the young, including women. PMID- 14746559 TI - A new Q/QS pattern on the resting electrocardiogram is associated with impaired insulin secretion and a poor prognosis in elderly men independently of history of myocardial infarction. AB - OBJECTIVES: To evaluate risk factors and prognosis of subjects who had developed a new Q/QS pattern on the resting electrocardiogram (ECG) in relation to history of myocardial infarction (MI). DESIGN: Cross-sectional and prospective population based cohort study. SETTING: Uppsala, Sweden. SUBJECTS: In 1970-73, all 50-year old men in Uppsala, were invited to participate in a health survey aimed at identifying risk factors for cardiovascular disease. The present study included the 1221 subjects who also were re-examined at age 70. RESULTS: Subjects with a new Q/QS pattern on the resting ECG at age 70 were characterized by impaired insulin secretion compared with those without Q/QS, and a higher prevalence of diabetes compared with the control group. In Cox proportional hazard analysis a new Q/QS pattern at age 70 was a significant predictor of cardiovascular mortality (hazard ratio : 1.67, 95% CI: 1.22-2.26) and total mortality (hazard ratio: 1.31, 95% CI: 1.04-1.62) (after age 70) during 9.4 years follow-up, also when adjusted for other risk factors and MI diagnosis. CONCLUSION: The finding of a new Q/QS pattern on the resting ECG, regardless of history of MI was associated with impaired insulin secretion and was an independent predictor of total and cardiovascular mortality. Therefore, these subjects must be given a high priority to preventive measures against both coronary heart disease and diabetes. PMID- 14746560 TI - Family burden of cardiovascular mortality: risk implications for offspring in a national register linkage study based upon the Malmo Preventive Project. AB - OBJECTIVE: To investigate the adjusted relative risk of cardiovascular disease (CVD) events in offspring of parents with cardiovascular mortality before 75 years. SETTING: The city of Malmo, Sweden. DESIGN: A follow-up study based on register linkage analyses. SUBJECTS AND METHODS: In the Malmo Preventive Project (MPP), a total of 22,444 men and 10,902 women attended the screening programme between 1974 and 1992. At the screening conventional risk factors for CVD were measured (blood pressure, lipids, glucose, smoking and social class). MAIN OUTCOME MEASURES: Parental CVD mortality was determined via register linkage analysis between the Multiple-Generation Register and the National Mortality Register (NMR). CVD events (morbidity and mortality) in offspring were collected from national registers. The relative risk for CVD events in offspring, in relation to parental CVD mortality, was adjusted for age and risk factors at screening. RESULTS: The age-adjusted relative risk (RR; 95%CI) for a son to experience a CVD event was increased in relation to a maternal positive family history of CVD mortality before 75 years when compared with no maternal history, RR 1.74 (1.43-2.11). This RR decreased to 1.51 (1.23-1.84; P < 0.001) after full adjustment for risk factors. The corresponding fully adjusted RRs for father-son heritage was RR 1.22 (1.02-1.47; P < 0.05), mother-daughter RR 0.87 (0.54-1.41), and father-daughter RR 1.20 (0.83-1.73). CONCLUSION: The existence of maternal CVD mortality before the age of 75 years implies a substantial risk increase for CVD morbidity and mortality in sons that cannot be explained by social background, lifestyle, or conventional cardiovascular risk factors in the adult offspring. PMID- 14746561 TI - Birth weight and cardiovascular risk factors in a cohort followed until 80 years of age: the study of men born in 1913. AB - OBJECTIVES: To analyse whether there is a relation-ship between birth weight and cardiovascular risk factors given the influence of potential modifying factors from birth time, former generations and adult life. DESIGN: Population-based cohort followed until 80 years of age. SETTING: Sweden. SUBJECTS: A total of 478 singleton men born in 1913 and participating in a population study in Gothenburg, Sweden, from age 50. MAIN OUTCOME MEASURES: Systolic blood pressure (SBP), antihypertensive treatment, incident diabetes mellitus, and serum total cholesterol, serum triglycerides and waist circumference as both continuous variables and in the highest quintiles of their distributions. RESULTS: After adjustment for the influence of birth time variables, hereditary factors and anthropometric and socio-economic adult life variables, SBP decreased by 3.7 mmHg per 1000 g increase of birth weight, the prevalence of antihypertensive treatment decreased by 32%, diabetes decreased by 53%, serum total cholesterol decreased by 0.20 mmol L(-1) and being in the top quintile of serum cholesterol decreased by 23%. The population risk percentage due to a birth weight < or =3000 g was for all three outcomes 3.8% and for antihypertensive treatment, diabetes and cholesterol 0.2, 18 and 2.5%, respectively. CONCLUSIONS: Low birth weight thus seems to affect the development of increasing SBP, antihypertensive treatment, diabetes and high cholesterol even when potential effect modifiers from birth time, former generations and adult life were taken into account. In the general population, the risk percentage due to a birth weight < or =3000 g was largest for diabetes. PMID- 14746562 TI - Microalbuminuria and stroke in a British population: the European Prospective Investigation into Cancer in Norfolk (EPIC-Norfolk) population study. AB - OBJECTIVES: To examine the relationship between microalbuminuria and incident stroke in the general population. DESIGN: Population-based prospective cohort study. SETTING: Participants were recruited in a primary care setting from 35 participating general practice units in Norfolk, UK. SUBJECTS AND MAIN OUTCOME MEASURES: The study population consisted of 23,630 individuals aged 40-79 years recruited between 1993 and 1997 for the EPIC-Norfolk Study and followed up for an average of 7.2 years. Random spot urine specimens were collected at baseline and albumin-to-creatinine ratio measured. Participants were categorized into normoalbuminuria, microalbuminuria and macroalbuminuria groups. During follow-up, the main end point was stroke incidence (fatal and nonfatal), ascertained from the UK Office for National Statistics and from the National Health Service Health District database of all hospital admissions. RESULTS: A total of 246 stroke events occurred during follow-up [crude incidence rate of stroke, 1.5 per 1000 person years (pyrs)]. The age-adjusted incidence of stroke increased significantly across categories of baseline albuminuria (0.9, 1.1 and 1.4/1000 pyrs for tertiles of normoalbuminuria, 2.6/1000 pyrs for microalbuminuria, and 6/1000 pyrs for macroalbuminuria in the total population, P < 0.001 for trend). In all women and men, the multivariate hazard ratio [95% confidence interval (CI)] for stroke associated with microalbuminuria was 1.49 (1.13-2.14) and macroalbuminuria 2.43 (1.11-6.26). After stratifying by stroke subtype, microalbuminuria was only independently predictive of ischaemic stroke, with hazard ratio (95% CI) of 2.01 (1.29-3.31). CONCLUSION: Microalbuminuria is independently associated with approximately 50% increased risk of stroke in the general population. Microalbuminuria may be useful in identifying those at increased risk of stroke in the general population. PMID- 14746563 TI - U-shaped relationship between mortality and admission blood pressure in patients with acute stroke. AB - OBJECTIVE: To evaluate the relationship between systolic blood pressure (SBP) or diastolic blood pressure (DBP) on admission and early or late mortality in patients with acute stroke. DESIGN: Prospective study of hospitalized first-ever stroke patients over 8 years. SETTING: Stroke unit and medical wards in a University hospital. SUBJECTS: A total of 1121 patients admitted within 24 h from stroke onset and followed up for 12 months. MAIN OUTCOME MEASURES: Mortality at 1 and 12 months after stroke in relation to admission SBP and DBP. RESULTS: Early and late mortality in patients with acute ischaemic or haemorrhagic stroke in relation to admission SBP and DBP followed a 'U-curve pattern'. After adjusting for known outcome predictors, the relative risk of 1-month and 1-year mortality associated with a 10-mmHg SBP increase above 130 mmHg (U-point of the curve) increased by 10.2% (95% CI: 4.2-16.6%) and 7.2% (95% CI: 2.2-12.3%), respectively. For every 10 mmHg SBP decrease, below the U-point, the relative risk of 1-month and 1-year mortality rose by 28.2% (95% CI: 8.6-51.3%) and 17.5% (95% CI: 3.1-34.0%), respectively. Low admission SBP-values were associated with heart failure (P < 0.001) and coronary artery disease (P = 0.006), whilst high values were associated with history of hypertension (P < 0.001) and lacunar stroke (P < 0.001). Death due to cerebral oedema was significantly (P = 0.005) more frequent in patients with high admission SBP-values, whereas death due to cardiovascular disease was more frequent (P = 0.004) in patients with low admission SBP-values. CONCLUSION: Acute ischaemic or haemorrhagic stroke patients with high and low admission BP-values have a higher early and late mortality. Coincidence of heart disease is associated with low initial BP-values. Death due to neurological damage from brain oedema is associated with high initial BP values. PMID- 14746564 TI - Changes in blood pressure and body weight following smoking cessation in women. AB - OBJECTIVE: Few have studied the long-term effects of smoking and smoking cessation on weight gain and blood pressure increase and compared with the age related increases experienced by most adults. This study compared the development of weight and blood pressure in female never smokers, continuing smokers and smokers who quit smoking. DESIGN: Weight, systolic (SBP) and diastolic (DBP) blood pressure and smoking habits were assessed at baseline and re-assessed after a mean follow-up of 9.0 +/- 5.8 years. SETTING: Population-based cohort. SUBJECTS: A total of 2381 female never smokers and 1550 female smokers. At the re examination, 388 of the smokers had quit smoking. RESULTS: Mean weight gain was 7.6 +/- 6.1, 3.2 +/- 5.8 and 3.7 +/- 5.2 kg, respectively, in quitters, continuing smokers and never smokers (P < 0.001). In women without blood pressure treatment, mean SBP increase was 20.9 +/- 16.8, 19.1 +/- 15.8 and 16.1 +/- 16.3 mmHg, respectively, in these groups (P < 0.001). Mean DBP increase was 6.2 +/- 8.7, 5.7 +/- 9.3 and 3.1 +/- 8.0 mmHg, respectively (P < 0.001). After adjustments for potential confounders, the increased weight gain in quitters remained highly significant. The differences in SBP and DBP increase were attenuated after adjustments, but remained significant. Incidence of hypertension (> or = 160/95 mmHg or treatment) was significantly higher in quitters [adjusted odds ratio (OR): 1.8; CI: 1.4-2.5] when compared with continuing smokers (OR: 1.3; CI: 1.07-1.6) and never smokers (reference). CONCLUSION: Over a long follow up, weight gain was approximately 3-4 kg higher in quitters when compared with continuing smokers or never smokers. Although the differences in blood pressure increase were moderate, smoking cessation was associated with an increased incidence of hypertension. PMID- 14746565 TI - Early alterations in the postprandial VLDL1 apoB-100 and apoB-48 metabolism in men with strong heredity for type 2 diabetes. AB - OBJECTIVES: To study the postprandial triglyceride-rich lipoprotein (TRL) metabolism, specifically the concentrations of very low-density lipoproteins (VLDL); from intestine (apoB-48) and liver (apoB-100), in men with normal fasting triglycerides but at increased risk of developing type 2 diabetes. DESIGN: Cross sectional study. SUBJECTS AND SETTINGS: Sixteen healthy men with at least two first-degree relatives with type 2 diabetes were individually matched with 16 control subjects without known diabetes heredity for: age, body mass index, and fasting triglyceride level. They underwent an 8-h meal tolerance test (919 kcal, 51 g fat) during which lipoproteins were separated by density gradient ultracentrifugation. They were characterized by euglycaemic hyperinsulinaemic clamp, peak VO2, 7-day diet registration and computed tomography. RESULTS: The relatives were, as expected, more insulin resistant than the controls and had increased concentration of postprandial VLDL1 particles (49% higher for VLDL1 apoB-48, P = 0.04 and 21% higher for VLDL1 apoB-100, P = 0.048). The elevation was related to insulin sensitivity, but not to lifestyle and body composition. Moreover, the concentration of postprandial triglycerides in VLDL1 fraction was inversely related to low-density lipoprotein (LDL) size in both relatives (rs = 0.60, P = 0.03) and controls (rs = -0.72, P = 0.004). There were no differences in the concentration of triglycerides or apoB-48 and apoB-100 particles in the other fractions (plasma, chylomicron or VLDL2). CONCLUSION: Increased postprandial concentration of TRLs in the VLDL1 fraction seems to be present at an early stage in the development of diabetes and probably contributes to the excess risk of future coronary events in insulin-resistant men. PMID- 14746566 TI - Is the relationship between type of alcohol and mortality influenced by socio economic status? AB - OBJECTIVES: To assess the impact of socio-economic status on the relationship between type of alcohol and all-cause mortality. DESIGN: A prospective population study. SETTING: The Copenhagen City Heart Study, Denmark. SUBJECTS: A total of 14,223 men and women participated in the first examination of The Copenhagen City Heart Study in 1976-1978. The participants were followed up until 18th of September 2001 during which 7208 persons died. The effect of beer, wine and spirits on mortality was stratified according to levels of education, income and cohabitation, and the association was examined after controlling for intake of the other types of alcohol, and for sex, smoking, physical activity and body mass index. MAIN OUTCOME MEASURES: Number and time of death from all causes. RESULTS: Consumers of wine were better educated and wealthier compared with beer and spirits drinkers. The association between type of beverage and mortality was noticed to differ according to socio-economic level, especially where the apparent protective effect of wine consumption tended to be strongest in the lower income and educational groups. CONCLUSIONS: This study finds the specific effects of beer, wine or spirits to moderately diverge in the socio-economic groups. Future studies addressing the association between the type of beverage and mortality may need to more thoroughly take socio-economic factors into account. PMID- 14746567 TI - Hodgkin's lymphoma, Epstein-Barr virus reactivation and fatal haemophagocytic syndrome. AB - Haemophagocytic syndrome is a serious disorder, often related to Epstein-Barr virus (EBV) or other infectious agents. Frequently an underlying immune abnormality or a T-cell lymphoma is present. The combination of haemophagocytosis and Hodgkin's lymphoma seems to be rare. A 70-year-old female with rheumatoid arthritis was admitted with constitutional symptoms, persistent fever, pancytopenia, deranged liver enzymes, lymphadenopathy and splenomegaly. A fatal coagulopathy supervened. The clinical picture and the bone marrow findings indicated a haemophagocytic syndrome and a lymph node biopsy disclosed an EBV positive Hodgkin's lymphoma. EBV serology pointed at viral reactivation and a high EBV DNA content was detected in serum by real-time quantitative PCR analysis (5.5 x 10(6) copies per mL). The case history is presented and the literature is reviewed. PMID- 14746568 TI - Acute myocardial infarction: an unusual culmination of rheumatic pancarditis and antiphospholipid antibody syndrome. AB - A 29-year-old man with history of antiphospholipid antibody syndrome (APS) and two prior episodes of acute rheumatic fever developed a third episode of acute rheumatic fever. This was complicated by acute myocardial infarction due to spontaneous arterial thrombosis of the left anterior descending and right coronary arteries. We postulate that rheumatic pancarditis created an inflammatory, thrombogenic environment that facilitated coronary artery thrombosis secondary to APS, a novel association. Our patient was left with significant left ventricular dysfunction at a young age, and cases such as this emphasize the need for early recognition and proper treatment of APS. Further, the pathogenesis of thrombosis in APS is incompletely understood, and the cellular and molecular basis for this, including the role of 'second hits', are areas needing further investigation. PMID- 14746569 TI - HDL deficiency and atherosclerosis: lessons from Tangier disease. PMID- 14746570 TI - Mild cobalamin deficiency associated with long-term metformin intake--reply. PMID- 14746571 TI - Effects of losartan on diabetic maculopathy in type 2 diabetic patients. PMID- 14746573 TI - Weight loss in obese diabetic and non-diabetic individuals and long-term diabetes outcomes--a systematic review. AB - The aim was to review the evidence for the long-term effects of weight loss on diabetes outcomes in obese people or for those at risk of developing type 2 diabetes. Current evidence is mostly based on short-term studies. This is a systematic review of long-term outcomes of weight loss in studies published between 1966 and 2001. Eleven long-term studies with a follow up of more than 2 years were included. Results show that those with diabetes who lost weight intentionally significantly reduced their mortality risks by 25%. Additionally, weight loss of 9-13 kg was most protective. Patients with the risk of developing diabetes due to either family history of diabetes or impaired glucose tolerance, saw a reduction in this risk. Those with large weight losses achievable with surgical interventions reduced their risk by at least 63%. Metabolic handling of glucose improved in 80% of those already with type 2 diabetes who lost weight. Based on one large study, intentional weight loss in obese patients appears to have a beneficial effect on mortality risk for those with type 2 diabetes. Clearly, further studies are needed to endorse this. The risk of developing diabetes being reduced by weight loss was shown in seven studies. However, the results were from studies with different analytical adjustments and outcome indices, making it difficult to make direct comparisons and should be viewed with caution. More long-term prospective studies need to be conducted with commitment to improving the methodological quality and standardization, in order to accurately assess the long-term effects of weight loss for obese diabetic and non diabetic individuals. PMID- 14746574 TI - Signalling pathways involved in retinal endothelial cell proliferation induced by advanced glycation end products: inhibitory effect of gliclazide. AB - AIM: We have previously demonstrated that advanced glycation end products (AGEs) stimulate bovine retinal endothelial cell (BREC) proliferation through induction of vascular endothelial growth factor (VEGF) production by these cells. We have also shown that gliclazide, a sulfonylurea which decreases oxidative stress, inhibits this effect. The aim of the present study was to characterize the signalling pathways involved in AGE-induced BREC proliferation and VEGF production and mediating the inhibitory effect of gliclazide on these biological events. METHODS: BRECs were treated or not treated with AGEs in the presence or absence of gliclazide, antioxidants, protein kinase C (PKC), mitogen-activated protein kinase (MAPK) or nuclear factor-kappaB (NF-kappaB) inhibitors. BREC proliferation was assessed by measuring [3H]-thymidine incorporation into DNA. Activation of PKC, MAPK and NF-kappaB signal transduction pathways and determination of VEGF expression were assessed by Western blot analysis using specific antibodies. MAPK activity was also determined by an in vitro kinase assay. RESULTS: Treatment of BRECs with AGEs significantly increased cell proliferation and VEGF expression. AGEs induced PKC-beta translocation, extracellular signal-regulated protein kinase 1/2 and NF-kappaB activation in these cells. Pharmacological inhibition of these signalling pathways abolished AGE effects on cell proliferation and VEGF expression. Exposure of BRECs to gliclazide or antioxidants such as vitamin E or N-acetyl-l-cysteine resulted in a significant decrease in AGE-induced activation of PKC-, MAPK- and NF-kappaB signalling pathways. CONCLUSIONS: Our results demonstrate the involvement of PKC, MAPK and NF-kappaB in AGE-induced BREC proliferation and VEGF expression. Gliclazide inhibits BREC proliferation by interfering with these intracellular signal transduction pathways. PMID- 14746575 TI - A placebo-controlled crossover study comparing the effects of nateglinide and glibenclamide on postprandial hyperglycaemia and hyperinsulinaemia in patients with type 2 diabetes. AB - AIM: This was a randomized, double blind, three period crossover study. The objective was to compare glucose, insulin and C-peptide 24 h profiles in patients with type 2 diabetes mellitus after dosing with nateglinide (given preprandially before three test meals), glibenclamide (administered once before breakfast) or placebo (given before three test meals). METHODS: Fourteen patients underwent screening followed within 3 weeks by three treatment periods of 1 day, each separated by 7 days. Dosing followed a six-sequence balanced, two 3 x 3 replicated Latin square. RESULTS: Mean peak serum insulin levels were lower after nateglinide (115 mU/l) than after glibenclamide (145 mU/l.h; p = 0.017) but higher than after placebo (79 mU/l; p = 0.001). However, peak insulin levels were reached earlier after nateglinide [mean time to peak (tmax) 1.7 h] compared to glibenclamide (mean tmax 2.1 h, p = 0.06). Total insulin exposure over the day was higher after glibenclamide compared with that following nateglinide (1216 vs. 1067 mU/l.h; p = 0.009). Similar findings were seen with serum C-peptide. Despite this, mean peak plasma glucose concentrations were lower following nateglinide (11.4 mmol/l from a baseline of 8.3 mmol/l) compared with glibenclamide (13.2 mmol/l from a baseline of 8.5 mmol/l; p = 0.001) and placebo (14.0 mmol/l from a baseline of 8.0 mmol/L; p < 0.001). CONCLUSIONS: Nateglinide improves early prandial measures of insulin and glucose response to a standard meal, more so than glibenclamide, in people with type 2 diabetes. PMID- 14746576 TI - Prevention of weight gain in type 2 diabetes requiring insulin treatment. AB - BACKGROUND: Patients with type 2 diabetes who are failing on oral agents will generally gain a large amount of body fat when switched to insulin treatment. This adverse effect may be related to chronic hyperinsulinism induced by long acting insulin compounds. AIM: To test the concept that regain of glycaemic control can be achieved without causing weight gain, using a regimen free of long acting insulin. METHODS: In a 3-month open-label pilot study including 25 patients with moderate overweight and secondary failure, we investigated whether nocturnal glycaemic control could be achieved with glimepiride administered at 20:00 hours. The starting dose was 1-2 mg, with subsequent titration up to a maximum of 6 mg. Rapid-acting insulin analogues were used three times daily to regain postprandial glucose control. RESULTS: Glycaemic control at 3 months was established with glimepiride in a dose of 4.4 +/- 0.3 mg/day (mean +/- standard error of the mean), and a total daily insulin dose of 24.1 +/- 2.6 IU. Fasting glucose levels decreased from 12.7 +/- 0.6 mmol/l to 8.1 +/- 0.3 mmol/l (p < 0.001), and target levels were reached in 14 of 25 patients (56%). Mean HbA1c decreased from 10.5 +/- 0.4 to 7.7 +/- 0.2% (p < 0.001). Symptomatic nocturnal hypoglycaemia was not reported. Body weight did not change (85.7 +/- 3.6 kg vs. 85.7 +/- 3.3 kg, p = 0.99). CONCLUSION: The data suggest that this new approach may be useful in about 50% of type 2 diabetes patients presenting with failure on maximal oral treatment. PMID- 14746577 TI - The effect of high-fat and high-fructose diets on glucose tolerance and plasma lipid and leptin levels in rats. AB - AIM: High-fat and high-fructose diets are usually used to induce animal model diabetes mellitus. The purposes of this research were to compare the abnormalities of glucose metabolism caused by high-fructose diet and a high-fat diet and the effects of the high-fructose diet and high-fat diet on plasma leptin. METHODS: In this research, 24 Sprague-Dawley rats were used as the experimental animals, which were divided into three groups: chow diet (control group), high-fructose diet (60% fructose w/w) and high-fat diet (20% lard w/w). They were fed for a period of 8 weeks, during which an oral glucose tolerance test was conducted in the seventh week, and after completion of the eighth week, the abdominal adipose tissue and liver of the rats were excised and weighed, and the plasma cholesterol, triglyceride, insulin and leptin concentrations were assayed. RESULTS: The high-fat diet group presented a fasting blood glucose concentration that was higher than that of the control group. Furthermore, after 2 h of glucose challenge, the rats in the high-fat and high-fructose diet groups all presented higher plasma glucose concentrations than did the control group. The high-fat diet group showed higher body weight, higher relative liver weight, a higher plasma cholesterol concentration and higher amylase activity than did the other groups, whereas the high-fructose diet group showed higher fasting insulin and triglyceride concentrations. As for adipose tissue, the high-fat diet group presented an amount that was higher than that of the high-fructose and control groups, but the plasma leptin concentration of the high-fructose group was higher than that of the control group. CONCLUSIONS: It can be concluded from the above-mentioned experimental results that a high-fructose diet can cause hyperinsulinaemia, while a high-fat diet can result in impaired pancreatic function of insulin secretion and glucose intolerance, indicating that high fructose diet and a high-fat diet may exert divergent effects on glucose metabolism in rats. PMID- 14746578 TI - The metabolic syndrome as a link between smoking and cardiovascular disease. AB - OBJECTIVE: Smoking is associated with a significant increase in the cardiovascular risk. The possible relationship of smoking with insulin resistance might further enhance the cardiovascular risk of the patients and is therefore of great clinical interest. DESIGN, SETTING AND SUBJECTS: We have retrospectively evaluated data of 3804 non-diabetic men attending a medical outdoor clinic. Clinical [body mass index (BMI), percentage of body fat, waist-to-hip ratio] and laboratory results were compared between smokers (n = 124) and non-smokers (n = 1915) without cardiovascular disease, as well as between smokers (n = 759) and non-smokers (n = 1006) with cardiovascular disease. RESULTS: Smokers without clinically manifest cardiovascular disease revealed significantly higher fasting glucose (5.8 +/- 0.6 mmol/l) and triglyceride levels (1.8 +/- 0.9 mmol/l) than non-smokers (fasting glucose: 5.1 +/- 0.7 mmol/l, p < 0.010; triglycerides: 1.5 +/- 0.8 mmol/l, p < 0.030). The adverse metabolic profile of smokers was even more pronounced in patients with cardiovascular disease. An age-matched analysis of smokers could demonstrate that cardiovascular patients revealed higher BMI values (27.3 +/- 2.4 kg/m2) and a higher percentage of body fat (25.5 +/- 5.5%) than those without cardiovascular disease (BMI: 25.7 +/- 2.2 kg/m2, p < 0.010; percentage of body fat: 23.0 +/- 5.5%, p < 0.030). CONCLUSION: In men with and without clinically manifest cardiovascular disease, smoking was associated with a metabolic profile indicating a higher degree of insulin resistance. PMID- 14746579 TI - The effects of oral anti-hyperglycaemic medications on serum lipid profiles in patients with type 2 diabetes. AB - AIM: Patients with type 2 diabetes often have dyslipidaemia, putting them at risk of cardiovascular disease, and are frequently treated with oral anti hyperglycaemic medications (OAMs). This review compares the effects of OAMs on serum lipids [total cholesterol (TC), low-density lipoprotein cholesterol (LDL C), high-density lipoprotein cholesterol (HDL-C), triglycerides (TGs) and free fatty acids (FFAs)] in patients with type 2 diabetes. METHODS: medline was searched for entries indexed from January 1966 to November 2002; search terms included the names of OAMs and serum lipids, limited to English language and human subjects. We selected clinical studies in type 2 diabetes of OAM monotherapy that included serum lipid data, treated all patients in a treatment group with the same drug, used therapeutic OAM doses not higher than the maximum recommended in the USA, compared therapy with baseline or placebo and specified statistical tests used. One unblinded investigator selected studies for inclusion. Data reported include number of patients, study length, OAM dose, serum lipid data at baseline and endpoint, p-values and statistical tests. RESULTS: Data on the serum lipid effects of sulphonylureas, repaglinide, nateglinide and miglitol were inconclusive. Acarbose increased HDL-C and decreased LDL-C and voglibose reduced TC. Metformin at higher doses reduced TC; data on its effects on other lipids were inconclusive. Rosiglitazone increased LDL-C, HDL-C and TC and reduced FFAs but had no effect on TGs. Pioglitazone increased HDL-C and reduced TGs and FFAs but did not affect LDL-C or TC. CONCLUSIONS: Lipid changes as a result of improved glycaemic control are not uniform findings associated with anti-diabetic therapy. Only metformin, acarbose, voglibose, rosiglitazone and pioglitazone had significant effects on the lipid profile. These effects should be considered when selecting OAMs for patients with type 2 diabetes. PMID- 14746580 TI - Whither the blade? PMID- 14746581 TI - Orbital floor blow-out fractures. PMID- 14746582 TI - Is directed donation misguided? AB - The worldwide shortage of organs and tissue for transplant has led to many ethical discussions involving restrictions concerning organ and tissue donation, including living donations and payment for donation. Efforts are being made to increase the donor pool; however, it is timely to ask whether moral decisions such as the rejection of directed donations are defensible. In this Clinical Conundrum, six specialists delve into the ethical and practical issues surrounding directed donation of human organs and tissues with particular reference to its implications in the field of ophthalmology. PMID- 14746583 TI - Major determinants of optic nerve head topographic characteristics in a normal Turkish population. AB - AIM: To determine the factors affecting the topographic characteristics of the optic nerve head in a normal Turkish population. METHODS: A cross-sectional study was performed on the optic nerve head of the right eyes of 613 healthy subjects of both sexes aged 11-77 years using a confocal scanning laser ophthalmoscope. The study population was divided into three groups (<20 years, 20-50 years, and>50 years) to evaluate age-related changes in the optic nerve head topography. A total of 15 variables were determined. Statistical analysis was performed using Student's t-test, anova, Tukey HSD test, linear regression analysis and Pearson's correlation coefficient. RESULTS: Four eyes were excluded from the study due to poor image quality. There were 307 female subjects and 302 male subjects. The mean age of the subjects was 42.1 +/- 15.1 years. The mean optic disc areas were 1.99 +/- 0.45 mm2 and 1.97 +/- 0.41 mm2 for male and female subjects, respectively. The disc area of the subjects ranged between 1.01 and 4.19 mm2. Disc area was found to be associated with all of the optic nerve head variables studied, and was also found to be significantly increased with advancing age (P < 0.05). The only variable that showed a sex-related difference was neuroretinal rim volume, which was found to be higher in female subjects in all cases. CONCLUSION: Optic disc area was found to be the main determinant of the optic nerve head topography in a normal population. Optic disc area was found to be significantly increased with ageing in a normal population. These acquired changes to the optic disc could suggest stretching of the scleral ring, which may highlight the role of advancing age on the pathogenesis of glaucomatous optic neuropathy. PMID- 14746584 TI - Primary infantile glaucoma in an Australian population. AB - BACKGROUND: Primary infantile glaucoma presents rarely, but can be responsible for significant visual morbidity. There is little information on the clinical features and visual outcome of a pure population of primary infantile glaucoma, as opposed to a mixed population of primary and secondary glaucoma or combined group of those with trabeculodysgenesis and iridotrabeculodysgenesis. METHODS: We conducted a retrospective review of children with primary infantile glaucoma seen in south-eastern Australia between 1980 and 2000, using The Royal Children's Hospital ophthalmic diagnostic coding database. RESULTS: Fifty-one patients with primary infantile glaucoma were identified (83 eyes). This equates to an estimated incidence of approximately 1 in 30,000 births. The mean +/- SD age at presentation was 135 +/- 84 days. 'Burnt-out' disease (megalocornea without raised intraocular pressure) was diagnosed in 10.8%. Goniotomy was the most commonly performed surgical procedure (69.4% of 72 eyes). Surgical success with one or two goniotomies was achieved in 74% of eyes. Visual outcomes at final review were generally good with 61.8% reading 6/12 or better. There were a disproportionately high number of children having a final recorded acuity of <6/60 in the group diagnosed in the first 3 months of life. CONCLUSIONS: Primary infantile glaucoma is a rare ocular condition in this population that presents at a mean age of 4.4 months. Surgical and visual outcomes are generally favourable. PMID- 14746585 TI - Outcome of consecutive trabeculectomy. AB - PURPOSE: To establish a criterion for success of primary phakic trabeculectomy in the second eye of the same patient, using the first operated eye as a predictor for the surgical outcome. METHODS: The outcome of primary phakic trabeculectomy was retrospectively compared in both eyes of 23 patients. Sixteen patients were treated with antimetabolites and seven were not. Postoperative intraocular pressure and number of glaucoma medications in paired eyes of the same patients were compared. Surgical success was defined as postoperative IOP of <20 mmHg without medication. Bleb morphology and the number of glaucoma medications were also compared in both eyes. RESULTS: There was a positive correlation between the two eyes of a patient in the IOP values at each postoperative time point in both groups (with and without antimetabolites) (P > 0.05). At the last follow-up visit after trabeculectomy, the number of glaucoma medications used in paired eyes were not significantly different (P > 0.83). Surgical failure occurred more often in paired eyes than in single eyes. Bilateral failure occurred in 60% (3/5) of the failed trabeculectomies with antimetabolites and in 100% (3/3) of the failed trabeculectomies without antimetabolites. CONCLUSION: Trabeculectomy outcome in paired eyes of patients was similar in both antimetabolites treated and untreated eyes. Thus, trabeculectomy outcome in the first operated eye can predict the surgical result in the second eye of the same patient. PMID- 14746586 TI - Long-term evaluation of trabeculectomy in primary open angle glaucoma and chronic primary angle closure glaucoma in an Asian population. AB - PURPOSE: A retrospective cohort study was undertaken to evaluate and compare the long-term results of trabeculectomy in primary open angle glaucoma (POAG) and chronic primary angle closure glaucoma (CPACG) in an Asian population. METHODS: Yearly diurnal measurements of intraocular pressure (IOP), best-corrected visual acuity, optic disc and visual field records of patients having primary adult glaucomas who had undergone trabeculectomy, without anti-mitotic agents, with a minimum of 5 years follow up were evaluated. Only one eye of each patient was studied. The success rates for IOP control in POAG and CPACG were statistically analysed. RESULTS: Sixty-four eyes of 64 patients were studied. The overall probability of success of trabeculectomy in controlling IOP to < or = 21 mmHg with or without additional topical antiglaucoma medication was 0.94 and 0.88 at 5 and 10 years, respectively. There was no statistically significant difference in the qualified and absolute success rates for IOP control between POAG and CPACG eyes (log rank test P= 0.6, 0.88, respectively). Twelve of 38 CPACG eyes had a two-line decrease in visual acuity as compared to four of 26 POAG eyes (P = 0.17). Progression or development of a cataract was the most common cause of visual decline. CONCLUSIONS: Trabeculectomy without antimetabolite use appears to be efficacious in lowering IOP and in visual field preservation over a period of 10 years in both POAG and CPACG. Development/progression of cataract especially in eyes with chronic angle closure glaucoma after trabeculectomy must be considered an important issue. PMID- 14746587 TI - Comparison of the Proview pressure phosphene tonometer performed by the patient and examiner with the Goldmann applanation tonometer. AB - OBJECTIVE: To compare the results of Proview pressure phosphene tonometry (PPPT) performed by the patient and an examiner with Goldmann applanation tonometry (GAT). METHODS: A comparative case series of 96 (192 eyes) consecutive patients from a glaucoma clinic was conducted. Intraocular pressure (IOP) was measured with GAT by one examiner, PPPT by another examiner, and PPPT by the patient. All examiners were masked to the results from any preceding IOP measurement. RESULTS: The coefficient of repeatability for repeated measurements for the GAT was +/ 0.48 mmHg. The limits of agreement for self-measurement of IOP with the PPPT and examiner measured IOP with PPPT were 6.3 mmHg and 4.8 mmHg, respectively. The limits of agreement between GAT and self assessed PPPT were +/-11.8 mmHg (mean difference of 0.63 mmHg). When the same comparison was made between GAT and examiner assessed PPPT, the results were limits of agreement of +/-10 mmHg (mean difference of 2.86 mmHg). No significant difference was identified in the agreement of the GAT and the PPPT when subanalysed for age of patient or diagnosis (P > 0.05). The limits of agreement between self-assessed IOP with the PPPT and the GAT were +/-8.2 for those with IOP <20 mmHg and +/-14.9 mmHg for those with IOP>20 mmHg. CONCLUSIONS: Poor agreement exists between IOP measured by GAT and PPPT measured by an examiner or by the patient. PMID- 14746588 TI - Evaluation of contact versus non-contact diode laser cyclophotocoagulation for refractory glaucomas using similar energy settings. AB - PURPOSE: To compare the efficacy of contact and non-contact techniques of diode transscleral cyclophotocoagulation (TSCPC) in patients with refractory glaucoma in a prospective randomized manner. METHODS: Sixty eyes with refractory glaucoma were randomly treated with either the contact or non-contact technique of diode TSCPC and followed up for a minimum of 12 months. A standard protocol of 1500 mW for 2 s per spot, with 40 spots over the entire 360 degrees circumference was followed for all eyes in both groups. RESULTS: The reduction of mean baseline intraocular pressure (IOP) from 44.5 +/- 7.4 mmHg to 14.8 +/- 1.8 mmHg with the contact mode was not significantly different from that in the non-contact mode of 41.9 +/- 8.8 mmHg reduced to 14.5 +/- 1.5 mmHg (P = 0.34). The probability of IOP survival (<22 mmHg) at 1 year using the Kaplan-Meier plot was better using the contact mode, although the results did not achieve statistical significance. However, the average repeat treatments per eye with the non-contact mode (1.6 +/- 0.7) was significantly higher than with the contact method (1.2 +/- 0.6; P < 0.05). CONCLUSION: Using an energy setting of 3 J per spot, both the contact and non-contact methods of laser delivery were found to be equally effective in lowering IOP in eyes with refractory glaucoma. PMID- 14746589 TI - Black eye drop bottle tips improve compliance. AB - AIM: To determine if dark coloured tips on eye drop bottles facilitate ocular therapy. METHODS: Eye drop bottles were modified by placing sterile black tape around the bottle tip. Patients on regular timolol therapy were asked to use bottles with the modified tip for one month. They then completed a questionnaire comparing the black tips with the standard tips on the bottle they normally use. RESULTS: Eighty-eight per cent of patients found the black tipped bottles easier to use than bottles with the standard tip. Sixty-eight per cent of patients had fewer occasions on which they needed to instil a second drop and 30% of patients touched their eyelid less when using the modified bottle tips. CONCLUSION: Black coloured bottle tips aid ocular therapy. They are easier to use, result in less contact with the eye on instillation and patients note a reduction in need for a second or additional drop. This is likely to improve compliance and reduce contamination. A change in manufacturing practise should be encouraged. PMID- 14746590 TI - Management of isolated orbital floor blow-out fractures: a survey of Australian and New Zealand oral and maxillofacial surgeons. AB - BACKGROUND: This is the first report of involvement of Australian and New Zealand oral and maxillofacial surgeons in the management of isolated orbital floor blow out fractures and was conducted to obtain comparisons with the results from a recent similar survey of British oral and maxillofacial surgeons. METHODS: A questionnaire survey was sent to all 113 practising members of the Australian and New Zealand Association of Oral and Maxillofacial Surgeons in April 2002 with a second mailout 1 month later. RESULTS: Sixty-nine per cent of the respondents were referred isolated orbital floor blow-out fractures for management, and just over half of these respondents estimated that 50% or more of the cases went to surgery. The materials most commonly used in orbital floor reconstruction were resorbable membrane for small defects and autologous bone for large defects. CONCLUSION: As in Britain, management of isolated orbital floor blow-out fractures comprises part of the surgical spectrum for many oral and maxillofacial surgeons in Australia and New Zealand. The management protocol was observed to be very similar between the two groups. PMID- 14746591 TI - Late reactivation of thyroid orbitopathy. AB - BACKGROUND: Thyroid orbitopathy is characterized by spontaneous remissions and exacerbations. Although data on the natural history are scarce, the active phase is usually self-limited, encompassing an average period of 18-36 months. Late reactivation, defined as active orbitopathy occurring after more than 5 years of quiescent disease, appears to be an uncommon and poorly documented event. The aim of the present study was to assess the incidence and characteristics of late reactivation of thyroid orbitopathy within a tertiary referral orbital clinic. METHODS: A retrospective case review was conducted of patients with thyroid orbitopathy seen at the Royal Adelaide Hospital Orbital Clinic between 1996 and 2001. RESULTS: From 193 cases of thyroid orbitopathy, eight cases (5%) of late reactivation were identified. The mean age of patients was 51 years (range 32-76 years) with a female to male ratio of 3:1. All cases presented with proptosis and seven demonstrated increased diplopia and gaze restriction. Signs of soft tissue inflammation were seen in three cases. All were euthyroid at presentation with a past history of thyroid orbitopathy and hyperthyroidism. One patient had an association of smoking with reactivation but the remainder had no discernible precipitants. The average interval between the initial orbitopathy and reactivation was 12 years (range 6-30 years). Computed tomography revealed enlarged extraocular muscles in seven cases and two showed partial response to treatment with low dose radiotherapy and steroids. The average length of disease activity was 14 months (range 9-18 months). CONCLUSION: Late reactivation of thyroid orbitopathy appears to be an uncommon phenomenon. It may occur under euthyroid conditions with no obvious precipitants and often presents as a reactivation of myopathy. PMID- 14746592 TI - Pattern of orbital exenteration in a tertiary eye care centre in India. AB - PURPOSE: This study aimed to determine the clinical indications for orbital exenteration, profile of these patients and clinicopathological correlations, and to compare these results with previous published data. METHODS: A retrospective analysis was conducted of exenterations performed in adults at a tertiary eye care centre in India over a period of 10 years (January 1990 to December 2000). Patient records were reviewed to obtain demographic data, presenting symptoms and their duration, laterality, and clinical and histopathological diagnosis. RESULTS: A total of 26 cases were identified. Patients ranged in age from 32 to 72 years (mean +/- SD 58.7 +/- 9.23 years). No sex predilection was observed. Classification of cases on histopathological criteria showed that exenterations were performed mostly for squamous cell carcinoma (10 cases), followed by sebaceous gland carcinoma (six cases) and basal cell carcinoma (two cases) of the eyelid. Conjunctival malignant melanoma contributed to 5 of 26 cases of exenteration. Adenoid cystic carcinoma of the lacrimal gland (one case), neurofibroma (one case) and orbital fungal infection (one case) were the other indications of exenteration. The clinicopathological correlation was 100% for squamous cell carcinoma, 100% for basal cell carcinoma, 80% for malignant melanoma and 75% for sebaceous gland carcinoma. CONCLUSION: Exenteration is mainly performed as a life-saving treatment for advanced malignant tumours with epithelial tumours being the commonest. In comparison to previous published data, the indications of surgery in India differed, as squamous cell carcinoma and sebaceous gland carcinoma were the commonest indications for exenteration. PMID- 14746593 TI - Intravitreal sodium fluorescein enhances visualization of clear vitreous during vitreous surgery for macular hole: a safety and efficacy study. AB - PURPOSE: To assess the safety and efficacy of sodium fluorescein (NaFl) as an aid for clear vitreous vitrectomy. METHODS: In 50 consecutive cases of macular hole surgery, NaFl 0.6% prepared from 20% injectable dye was injected into the vitreous cavity after core vitrectomy so as to stain the clear formed vitreous. RESULTS: NaFl stained the peripheral uncut clear vitreous green and improved the visualization for a safe and complete vitrectomy. No complications associated with the dye injection were noted. CONCLUSION: NaFl 0.6% can safely be used in the vitreous cavity for easy identification of clear uncut vitreous gel for clear vitreous vitrectomy. This enhances expeditious excision of this tissue. PMID- 14746594 TI - Role of prophylactic scleral buckling in the management of retained intraocular foreign bodies. AB - PURPOSE: To evaluate the role of prophylactic scleral buckling procedure with pars plana vitrectomy for the removal of retained intraocular foreign bodies (RIOFB) in the prevention of postoperative retinal detachment. METHODS: In a prospective, clinical, controlled study, 28 patients with RIOFB without retinal detachment were randomly divided into two groups of 15 (group I) and 13 (group II) patients. All patients underwent a standard three-port pars plana vitrectomy with foreign body removal. In addition to this standard procedure, patients assigned to group I had a 360 degrees encircling scleral buckle placed at the time of surgery. Group II patients did not have any scleral buckling. All patients were followed up for an average period of 11.8 months. Both groups were comparable with respect to age, sex, duration of injury, nature of injury, and site, size and type of RIOFB. RESULTS: The retinal detachment rate was found to be 6.6% in group I and 30.8% in group II (P = 0.24). Prophylactic scleral buckling reduced the risk of retinal detachment by 24% but this was not found to be statistically significant. CONCLUSION: Prophylactic scleral buckle placement with pars plana vitreous surgery for RIOFB may reduce the risk of late onset retinal detachment. A larger study would seem warranted. PMID- 14746595 TI - Effects of topical antiglaucoma drugs on apoptosis rates of conjunctival epithelial cells in glaucoma patients. AB - BACKGROUND: To investigate the effects of topical antiglaucoma drugs on apoptosis rates of conjunctival epithelial cells in vivo. METHODS: Specimens were obtained from the bulbar conjunctiva of 30 patients who underwent either cataract or glaucoma surgery. Of these 30 patients, 14 had glaucoma and received topical antiglaucoma drugs. The 16 patients in the control group had neither systemic nor ocular disease except cataract and they did not receive any medications. TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling) method was used to show the apoptosis of the conjunctival epithelial cells. RESULTS: Increased apoptosis rates were detected in specimens of patients who were on topical antiglaucoma therapy (mean +/- SD 0.208 +/- 0.154) when compared with the healthy subjects (mean +/- SD 0.083 +/- 0.102; P = 0.013). The apoptosis rates did not differ according to type, number or duration of drugs. CONCLUSIONS: The apoptosis observed in the conjunctival epithelial cells might be due to the preservative benzalkonium chloride in the antiglaucoma medications because the apoptosis rates did not differ in patients using different types of medications. PMID- 14746596 TI - Controlled trial of hyperbaric oxygen treatment for alkali corneal burn in the rabbit. AB - PURPOSE: To evaluate the efficacy of hyperbaric oxygen therapy in the treatment of alkali-induced corneal burns in an animal model. METHODS: Twenty-four rabbits were randomized into a control group (n = 12) and hyperbaric oxygen treatment group (n = 12). After induction of anaesthesia, the alkali burn model was established by application of 1 N sodium hydroxide to one eye of each rabbit. The hyperbaric oxygen treatment group was treated each day for 21 days with hyperbaric oxygen at 2.4 Atmospheres Absolute (ATA) for 1 h. The eyes of the animals were examined daily for 2 weeks and then weekly until the end of the trial. The principal endpoint was that of perforation of the cornea at which time the animals were killed with a lethal dose of either intravenous or intraperitoneal barbiturate and the eyes immediately enucleated and fixed in 10% neutral buffered formalin. All animals in which complete healing took placed were also killed, the eyes removed, fixed and examined histologically. Photographs were taken of the rabbit's eyes at weekly intervals and the area of vascularization and epithelial defects in the hyperbaric and control groups were compared. RESULTS: Equal numbers (seven) of the control and hyperbaric oxygen treated groups had perforated corneas and there was no statistical difference in the mean time to perforation (control 30.1 days; treated 30 days). There was also no statistical difference between the two groups with respect to epithelial defect size. CONCLUSION: Treatment with hyperbaric oxygen for 1 h daily for 21 days had no beneficial effect on alkali-induced corneal burns. PMID- 14746597 TI - Arthur Thomas Paterson: the life and times of a New Zealand ophthalmologist. AB - Arthur Thomas Paterson was a pre-eminent New Zealand ophthalmologist who settled in the United Kingdom after the First World War and became a leading figure in the development of ophthalmic services in the north-east of England. This vignette explores his contributions to clinical education and the establishment of comprehensive ophthalmic services in Newcastle upon Tyne. It also highlights the Arthur Thomas Paterson scholarship bequest to enable New Zealand ophthalmologists to acquire additional specialist training in inter-national centres of excellence. PMID- 14746598 TI - Goldenhar syndrome: a case from Papua New Guinea. AB - Goldenhar syndrome is well known for its classical triad of epibulbar dermoids or lipodermoids, auricular appendages and pretragal fistulas. The syndrome was later included under the broader classification of oculoauriculovertebral spectrum to cover a wide range of associated anomalies. Here, a case is presented of Goldenhar syndrome in a child from Papua New Guinea. The patient had large epibulbar lipodermoids requiring surgical debulking primarily for mechanical lagophthalmos and secondarily for cosmesis. Further multidisciplinary effort is required to manage her systemic manifestations of the syndrome. PMID- 14746599 TI - Fluorescein angiographic monitoring of corneal vascularization in lipid keratopathy. AB - The use of digital fluorescein corneal angiography to assist argon laser photocoagulation is reported. Photocoagulation was performed on the vascular supply of lipid keratopathy in the left eye of a 44-year-old woman. PMID- 14746600 TI - Systemic anti-inflammatory fibrosis suppression in threatened trabeculectomy failure. AB - PURPOSE: To provide a rationale for the use of systemic anti-inflammatory fibrosis suppression in the postoperative management of threatened early trabeculectomy bleb failure. METHODS: A review of the literature and of the authors' own experiences was conducted. RESULTS: The most important cause of persistent elevation of intraocular pressure after trabeculectomy is unduly marked or persistent inflammation with deposition of fibrous tissue, which prevents the formation of an adequately draining bleb. It was found that a clinically useful degree of suppression of bleb inflammation and fibrosis can be obtained with a 4-6 week course of the combined systemic administration of prednisone (10 mg t.i.d.), a non-steroidal anti-inflammatory agent (e.g. diclofenac 100 mg SR daily) and colchicine (0.25 mg or 0.3 mg t.i.d.), which was termed anti-inflammatory fibrosis suppression. Topical atropine 1% t.i.d. and adrenaline 1% t.i.d. eye drops can also be considered in addition to routine postoperative topical steroids. CONCLUSIONS: Despite advances in surgical techniques and methods to control fibrosis, anti-inflammatory fibrosis suppression is a valuable tool to have available in the post-operative period for management of trabeculectomies that threaten failure. This regime produces a diffuse bleb, which has a very low risk of late infection or bleb perforation. It is recommend that this regime be added to the list of therapies that are considered when clinical features suggestive of a failing bleb are confronted early in the post-operative course. PMID- 14746601 TI - Review of severe vaso-occlusive retinopathy in systemic lupus erythematosus and the antiphospholipid syndrome: associations, visual outcomes, complications and treatment. AB - PURPOSE: To discuss the pathogenesis of severe vaso-occlusive retinopathy in systemic lupus erythematosus (SLE), the association with antiphospholipid antibodies, and its implications for management and prognosis. METHODS: An illustrative case history of a woman with SLE and severe vaso-occlusive retinopathy in the presence of antiphospholipid antibodies is presented. A literature review of previously reported cases and previously published data on the topic was performed and forms the basis for discussion. RESULTS: This is a rare form of retinopathy in SLE as distinct from the more common, benign form, being classically a microangiopathy with diffuse capillary non-perfusion and small arterial or arteriolar occlusions in the retina. Poor visual outcomes with visual loss are reported in 80% of cases with neovascularization occurring in 40% of cases. It is associated with antiphospholipid antibodies, typically characterized by microthrombosis and immune complex mediated vasculopathy rather than a true vasculitis. There is a strong association between this severe form of retinopathy and central nervous system manifestations of SLE. Anticoagulation has a role in the secondary prevention of thrombosis in the presence of antiphospholipid antibodies, but the role of aspirin and immunosuppression is unclear in the treatment of this condition. Vigilant ophthalmic follow up and aggressive treatment of neovascularization and vitreous haemorrhage can prevent further visual loss. These points are highlighted in the brief case report presented. CONCLUSION: Severe vaso-occlusive retinopathy is a rare form of retinopathy in SLE often associated with poor visual prognosis and neovascularization. It may be a manifestation of the antiphospholipid syndrome. Treatment is aimed at preventing further thrombosis and complications arising from neovascularization. PMID- 14746602 TI - Retinoblastoma, microphthalmia and the chromosome 13q deletion syndrome. AB - A 10-month-old boy, known to have the 13q deletion syndrome, presented with leukocoria in a microphthalmic right eye. He was found to have bilateral retinoblastoma requiring enucleation of the right eye and laser, cryotherapy and external beam radiation to the left eye. This case demonstrates that retinoblastoma may be present in a microphthalmic eye and that more than one imaging modality should be used in the evaluation of suspected retinoblastoma, as calcification may be absent on the computed tomography scan but present on B-mode ultrasound. Children with a known 13q deletion require immediate and serial ophthalmological examinations. PMID- 14746603 TI - Malignant melanoma in eviscerated eyeball. AB - A 65-year-old woman presented with malignant melanoma of the right socket following evisceration of her right eye for ocular trauma 8 years previously. Computed tomography demonstrated a solid mass adjacent to the residual eviscerated eyeball without extra-orbital extension. The tumour was removed by total exenteration of the orbit. The clinical and radiological features of this condition, and a review of the literature are presented. PMID- 14746604 TI - Tamoxifen optic neuropathy. AB - A 68-year-old woman with metastatic breast cancer presented with slowly progressive visual loss in each eye. Electrophysiology confirmed bilateral optic neuropathies. Investigations to determine the cause of the vision loss were negative. Cessation of tamoxifen resulted in a dramatic improvement in acuity. PMID- 14746605 TI - Propecia-associated bilateral cataract. AB - A 43-year-old man presented suffering from decreasing vision in both eyes for 3 months. The patient's visual acuity was 6/20 (non-corrected) in the right eye and 6/10 (-1.75/-1.00 x 91) in the left. Ocular examination of both eyes revealed anterior subcapsular opacities of both lenses with the right eye being more severe than the left. He had been taking finasteride (Propecia; Merck, Sharp and Dohme) at 1 mg/day for 3 years to treat early stage of androgenic alopecia. It was highly suspected that finasteride was associated with the anterior subcapsular opacity on the lens, and the patient therefore discontinued use of finasteride. He underwent uneventful cataract extraction surgery and intraocular lens implantation of the right eye. One month after cataract surgery in the right eye, the best-corrected visual acuity was right 6/6 (-1.25) and left 6/10 (-2.00/ 0.50 x 100). To the best of the authors' knowledge, this is the first reported case of Propecia-associated cataract. PMID- 14746606 TI - Castleman's disease of the lacrimal gland. AB - Castleman's disease, also known as angiofollicular lymphoid hyperplasia, is a lymphoproliferative disorder that is generally benign. The lesions most commonly originate in the lymph nodes of the mediastinum. The second documented case of Castleman's disease of the lacrimal gland is reported in an 84-year-old woman who presented with an 8-year history of a left upper lid mass. This was excised via a lateral orbitotomy, and a diagnosis of Castleman's disease was made histologically. Relevant images of the clinical findings and histology are shown. PMID- 14746607 TI - Lipoid proteinosis: a rare disorder with pathognomonic lid lesions. AB - Lipoid proteinosis is a rare disorder seen even more rarely by ophthalmologists. The interesting clinical and histopathological features of this disease are described in a brother and sister who presented predominantly with lid lesions, without any systemic complaints. Knowledge of the typical lid lesions may help to diagnose this disorder. PMID- 14746608 TI - Intraorbital ophthalmic artery aneurysms. AB - Angiographically documented aneurysms of the intraorbital ophthalmic artery are extremely rare. Two cases are presented of intra-orbital ophthalmic artery aneurysms, both associated with arteriovenous malformation (AVM). There have only been isolated reports documenting this combination of vascular anomalies. In the first case, the aneurysm was treated conservatively and in the second case, after staging angiography and interventional embolization, the aneurysm was removed along with the associated AVM. The clinical presentation, pathogenesis and management of this rare combination of vascular anomalies is briefly discussed. PMID- 14746609 TI - Optical coherence tomography of adult-onset fovemacular vitelliform dystrophy. AB - Adult-onset foveomacular vitelliform dystrophy is characterized by a focal, round or oval shaped, subretinal foveal yellow lesion that presents bilaterally and usually symmetrically in the fourth or fifth decade of life. In addition to clinical observation, diagnosis of the disease process has until now been assisted by electrophysiological and fundus fluorescein studies. Two cases are presented that correlate the clinical fundus appearance with optical coherence tomography findings and describe the location of the yellow vitelliform material. PMID- 14746610 TI - Comment on 'Macular hole surgery with or without indocyanine green stained internal limiting membrane peeling'. PMID- 14746611 TI - Comment on 'Photodynamic therapy in practice'. PMID- 14746612 TI - Cutaneous complications of intravenous drug abuse. AB - Injection drug abuse is a world-wide problem responsible for numerous minor to life-threatening and fatal complications. The skin is the tissue most evidently affected by intravenous drug addiction. A wide spectrum of cutaneous complications may occur in intravenous drug users. These include acute or delayed local complications, hypersensitivity reactions, cutaneous manifestations of systemic infections or becoming the site of toxigenic infections. Between 1996 and 2001, in our institution in south-eastern France, we observed cutaneous complications after crushed buprenorphine tablet injections in 13 patients. This paper reviews and classifies adverse effects of parenteral drug abuse on the skin. PMID- 14746613 TI - Updates from the British Association of Dermatologists 83rd annual meeting, 1-4 July 2003, Brighton, U.K. PMID- 14746614 TI - A new analytical system for quantification scratching behaviour in mice. AB - BACKGROUND: Scratching behaviour is an important component of human atopic dermatitis. The duration of scratching determines the extent of skin damage and thus the rash, but quantification of this is difficult. Establishment of a method for measuring the duration of scratching is important in order to make objective assessments of the factors that may cause the itch and also the efficacy of new antipruritic drugs. OBJECTIVES: A novel method for assessing the duration of scratching in mice was evaluated, based on the time course changes in the distance between the animal's hind limbs and the back of the neck during scratching behaviour. METHODS: Compound 48/80 was administered intradermally to the back of ICR mice and their scratching behaviour was recorded on digital videotape. The distance between the back and the hind limb was measured continuously using an image analysis system. RESULTS: Measurement of the frequency and duration when the mouse's hind limb came into contact with the back of the neck provided an accurate method of quantitating scratching behaviour. CONCLUSIONS: This system provides a new method of quantifying scratching behaviour in a mouse. PMID- 14746615 TI - Pigment-lightening effect of N,N'-dilinoleylcystamine on human melanoma cells. AB - BACKGROUND: Cystamine and linoleic acid have been reported to reduce melanin synthesis in vitro and in vivo. N,N'-dilinoleylcystamine (DLC) is a compound of cystamine and linoleic acid connected by an ester bond. OBJECTIVES: To investigate the effects of DLC on melanin synthesis using cultured human melanoma cells. METHODS: Levels of total melanin, eumelanin and phaeomelanin, tyrosinase protein and tyrosinase activity in situ were measured in HM3KO melanoma cells. Changes in degree of pigmentation were quantified by image analysis and compared with absorbance values. Tyrosinase from HM3KO cells was used to measure the direct effect of DLC on DOPA and DOPAchrome production. RESULTS: At concentrations of 1.4-14 micromol L-1, DLC reduced the pigmentation of HM3KO melanoma cells but did not affect cell growth. The visual decrease in pigmentation produced by DLC was more dramatic than the decrease in total melanin content as measured by absorbance at 500 nm. DLC treatment decreased eumelanin synthesis and increased phaeomelanin synthesis in HM3KO melanoma cells. An in situ tyrosinase assay showed that DLC inhibited tyrosinase activity, as well as the level of tyrosinase protein. CONCLUSIONS: These results suggest that DLC has pigment-lightening effects on HM3KO melanoma cells, produced by reducing the level of eumelanin while increasing the level of phaeomelanin. PMID- 14746616 TI - Characterization of the ultraviolet B and X-ray response of primary cultured epidermal cells from patients with disseminated superficial actinic porokeratosis. AB - BACKGROUND: Disseminated superficial actinic porokeratosis (DSAP) is the most common porokeratosis and is characterized by multiple keratotic lesions which tend to occur at sun-exposed sites. A mild hypersensitivity to X-rays has been reported for DSAP-derived fibroblasts and frequent over-expression of p53 has been found in lesional epidermis. OBJECTIVES: In order to clarify whether genome maintenance mechanisms might be compromised in this disease the following approaches were undertaken: (i) primary cultured keratinocytes and fibroblasts from DSAP patients were characterized for ultraviolet (UV) B and X-ray response; (ii) 15 lesions were studied for p53 mutations, and (iii) the differentiation status of DSAP-derived keratinocytes was evaluated. METHODS: Primary cultures of keratinocytes and fibroblasts were established from lesional and nonlesional skin biopsies of two subjects with DSAP. p53 mutations were analysed by DNA sequencing of the conserved region of the TP53 gene. Differentiation was evaluated both in stratified epithelial sheets from confluent keratinocyte cultures and in organotypic skin cultures. RESULTS: The cytotoxic and apoptotic response to UVB or X-irradiation was similar in DSAP-derived keratinocytes and fibroblasts when compared with normal cells. Two of 15 lesions examined presented p53 mutations located at nondipyrimidine sites. A strikingly decreased expression of filaggrin was observed both in reconstructed epidermis and in reconstructed skin. CONCLUSIONS: The UVB and X-ray response of DSAP-derived keratinocytes and fibroblasts indicates that the actinic character of this skin pathology is not due to radiation hypersensitivity. In agreement with this finding, mutations in the p53 gene, which are often associated with UV-related skin carcinogenesis, were rarely detected in DSAP lesions and were not UV-specific. Reconstructed epidermis and reconstructed skin models successfully reproduced the main features of this genodermatosis, showing that DSAP-derived keratinocytes bear an inherent defect in the terminal differentiation programme. PMID- 14746617 TI - Ageing of human epidermis: the role of apoptosis, Fas and telomerase. AB - BACKGROUND: Aged human epidermis is characterized by morphological changes including flattening of the dermal-epidermal junction and a decrease in thickness. OBJECTIVES: To determine the roles of proliferation, apoptosis, Fas (CD95), Fas ligand (FasL) and telomerase in changes of human epidermis during ageing. METHODS: Human epidermis from aged subjects (n = 14; mean age 70.7 years) and young subjects (n = 14; mean age 23.4 years) was studied by histology, immunohistochemistry, terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labelling assay for apoptotic cells and reverse transcription-polymerase chain reaction to determine epidermal thickness, proliferation (Ki-67), apoptosis, expression of Fas and FasL, and telomerase activity. RESULTS: Aged skin was associated with thinning of the epidermis, decreased proliferation, and increased apoptosis below the granular layer. This was associated with increased epidermal expression of Fas and FasL. Telomerase activity was similar in aged and young epidermis. CONCLUSIONS: Fas/FasL-mediated apoptosis, along with decreased proliferation, may have a role in changes of human epidermis during ageing. Telomerase activity did not appear to be limiting in young vs. old human epidermis. PMID- 14746618 TI - Clinically equivocal melanocytic skin lesions with features of regression: a dermoscopic-pathological study. AB - BACKGROUND: Benign melanocytic skin lesions may be difficult to differentiate from melanoma both clinically and dermoscopically. One of the most confounding dermoscopic features, commonly seen in melanoma but in our experience also in melanocytic naevi, is represented by the so-called blue-white structures (BWS). OBJECTIVES: To evaluate diagnostic significance and histopathological correlates of BWS seen by dermoscopy in a series of clinically equivocal melanocytic skin lesions that were excised. METHODS: Patients were recruited from six specialized pigmented lesion clinics in Austria, Italy and Spain over a period of 9 months. All consecutive patients showing one or more melanocytic lesions with BWS, but not classified as melanoma dermoscopically, were included. Each lesion was photographed clinically and dermoscopically. All images were reviewed by one of us and the degree, type and location of BWS evaluated for each lesion. A panel of four experienced dermatopathologists independently reviewed all specimens for diagnosis and one of them evaluated presence and degree of melanosis and/or fibrosis. The main outcome measures were the percentage and histopathological correlates of lesions with different degree, type and location of BWS. RESULTS: All included lesions with BWS (n = 158) showed partial or focal regression histopathologically. One hundred and thirty-five (85.4%) lesions were diagnosed as melanocytic naevi (complete histopathological interobserver agreement), whereas 23 (14.6%) were defined as equivocal because at least one of four pathologists diagnosed the given lesion as melanoma. Only one lesion was diagnosed as melanoma by all four pathologists. The majority of naevi exhibited blue areas (84.4%) with a central distribution (57%) and involving < 50% of the lesion surface (89.6%). By contrast, 78.3% of equivocal lesions revealed a combination of white and blue areas with an irregular distribution (60.9%) and involving > 50% of the lesion surface (47.8%). CONCLUSIONS: Using degree and type of BWS, an algorithm was constructed that can be applied for the management of lesions exhibiting dermoscopic features of regression. PMID- 14746619 TI - Inflammation in acne scarring: a comparison of the responses in lesions from patients prone and not prone to scar. AB - BACKGROUND: Many patients with inflammatory acne suffer from significant scarring, which is disfiguring and difficult to treat. A cell-mediated immune response is considered to be involved in the pathogenesis of acne, although the extent of this response has been found to differ among patients. OBJECTIVE: To assess whether there were differences in the cell-mediated immune responses at different time points in inflamed lesion development and resolution in patients who were prone (S patients) and those with the same degree of inflamed acne who were not prone (NS patients) to develop scarring. METHODS: Cellular and vascular markers were investigated using standard immunohistochemical techniques on biopsies of inflamed lesions of known duration, i.e. < 6 h (n = 14), 24 h (n = 14), 48 h (n = 10), 72 h (n = 10) and 6-7 days (n = 11) from the backs of acne patients. RESULTS: In early lesions from NS patients there was a large influx of CD4+ T cells, macrophages and Langerhans cells with a high number of cells expressing HLA-DR. Also there was significant angiogenesis and vascular adhesion molecule expression. Cell recruitment peaked in 48 h lesions, after which leucocyte numbers decreased and vascular activity returned to normal. Of the T cells, only 50% were memory/effector (CD45RO+) and naive (CD45RA+) cells, while the remainder were unclassified (CD45RO-, CD45RA-). In early lesions from S patients, CD4+ T cell numbers were smaller, although a high proportion were skin homing memory/effector cells. Langerhans cell numbers and cellular HLA-DR expression were low, while numbers of macrophages, blood vessels and vascular adhesion molecules were high. In resolving lesions angiogenesis remained high, with a further influx of macrophages and skin homing memory/effector cells and increased cellular HLA-DR expression. CONCLUSIONS: The cellular infiltrate was large and active with a greater nonspecific response (few memory T cells) in early lesions of NS patients, which subsided in resolution. In contrast, a predominantly specific immune response was present in S patients, which was initially smaller and ineffective, but was increased and activated in resolving lesions. Such excessive inflammation in healing tissue is conducive to scarring and suggests that the use of topical anti-inflammatory treatments would be appropriate for these patients. PMID- 14746620 TI - Artificial disruption of skin barrier prior to irritant patch testing does not improve test design. AB - BACKGROUND: Irritant patch testing is often performed as a 24- or 48-h occlusive patch test with low concentrations of sodium lauryl sulphate (SLS). OBJECTIVES: The aim of this study was to investigate potential ways to shorten this test procedure and obtain precise test results. PATIENTS AND METHODS: Thirty-six healthy volunteers underwent irritant patch testing with different pretreatments (PT) of the test fields. Occlusive test chambers were applied on the upper back with SLS 0.5%, 1%, 2% and 5% in large Finn Chambers(R). The patches were removed after 4 and 24 h, respectively, depending on the concentration used. Test fields were pretreated as follows: PT 0, field without any PT (control); PT 1, prick with lancet; PT 2, prick with test stamp; PT 3, scratch with lancet; PT 4, incision with standardized incision instrument (0.1-0.2 mm depth). Skin reactions were evaluated by transepidermal water loss (TEWL), skin erythema and skin hydration and as well by a visual score (VS) at 4, 24 and 72 h. RESULTS: Our data show an obvious distinction between PT 0-2 and PT 3-4 at all measurement methods. The average TEWL values with PT 3-4 were higher than those with PT 0-2, especially on the 4-h course. This distinction may derive from the shape and size of the skin impairment achieved by PT 3-4, leading to a mechanical barrier disruption. However, SLS may infiltrate directly into deeper skin layers supported by capillarity. Consequently, no or little penetration through the epidermis and interaction with its structures occurs, which is responsible for irritant skin reactions. The SLS dose in the upper skin layers is therefore lower at these PTs. The lower remaining dose of SLS also explains this distinction, especially for the VS. Additionally, there are presumed reactions in deeper layers of the epidermis and dermis at PT 3-4. CONCLUSIONS: In summary, all data suggest a different reaction pattern from the classical irritant response. Therefore, application without any PT seems to be best suited for irritancy skin testing, especially for visual assessment. PTs prior to irritant patch testing have been shown to be unjustifiable. PMID- 14746621 TI - Methyldibromoglutaronitrile in rinse-off products causes allergic contact dermatitis: an experimental study. AB - BACKGROUND: The frequency of sensitivity to the cosmetic preservative methyldibromoglutaronitrile (MDBGN) has increased significantly in Europe. Most cases of allergic contact dermatitis from MDBGN are caused by leave-on cosmetic products. The risk of developing allergic contact dermatitis from rinse-off products has been less studied. OBJECTIVES: To investigate the allergic response elicited in presensitized individuals from exposure to a rinse-off product preserved with the maximum permitted level of MDBGN. METHODS: Nineteen contact allergic individuals and nine controls participated in a double-blind, randomized repeated open application test (ROAT) using two coded liquid soaps with and without MDBGN. Areas of 50 cm2 on the lower arms were washed with the soaps twice a day for up to 28 days; two of the subjects continued for 34 days. The subjects were also patch tested with a dilution series of MDBGN to determine their patch test threshold values. RESULTS: Seven presensitized individuals (37%) developed allergic contact dermatitis from the soap containing MDBGN. The mean dose of MDBGN per application was 2.2 micro g cm-2 and the reactions appeared between days 6 and 34. All nine controls had negative ROATs. The difference in reactivity between test subjects and controls was significant (one-sided Fisher's exact test, P = 0.04). Patch test threshold values ranged from < 0.001% to 0.2% MDBGN in ethanol/water. CONCLUSIONS: This study shows that the exposure to a rinse-off product containing the maximum permitted level of MDBGN can easily elicit an allergic response in presensitized individuals. Along with reported cases of induction and elicitation caused by MDBGN in rinse-off products the study indicates that the permitted level of MDBGN in rinse-off products is too high. We recommend that this level should be re-evaluated. PMID- 14746623 TI - Prognosis and prognostic factors in adult patients with atopic dermatitis: a long term follow-up questionnaire study. AB - BACKGROUND: Atopic dermatitis (AD) is a chronic relapsing skin disease. Several investigations concerning the long-term prognosis of AD among children and teenagers have been performed but there are only few data among adults. OBJECTIVES: To investigate the prognosis and prognostic factors in adult patients with AD by a long-term follow-up (25-38 years). The prognostic factors were defined as those factors of importance for the persistence of AD. PATIENTS AND METHODS: A follow-up questionnaire was sent in November/December 1998 to 922 AD patients examined in our outpatient clinic between 1960 and 1973 among 1366 registered patients with AD. The patients were aged 20 years or older when they visited the clinic and 45 years or older when they answered the follow-up questionnaire. RESULTS: The response rate was 90.4%. The age range at the time of follow-up was 45-86 years (mean 55 years). Of the 833 patients who responded, 59% reported AD at some time during the last 12 months, which we defined as persistent AD. The mean value of clearance rate per person-years was 18%. One of the most important factors associated with persistence of AD was a head and neck dermatitis with or without other AD locations at the time of examination according to the old patient records. CONCLUSIONS: This study showed that the majority of adults with AD still had AD when they became older. This applies particularly if negative prognostic factors existed. PMID- 14746622 TI - Validation of the Eczema Area and Severity Index for atopic dermatitis in a cohort of 1550 patients from the pimecrolimus cream 1% randomized controlled clinical trials programme. AB - OBJECTIVE: To validate the Eczema Area and Severity Index (EASI) by assessing its internal consistency, reliability and sensitivity to change and by correlating it to other efficacy parameters. DESIGN: Three short-term and two long-term double blind, randomized, controlled trials, performed in 138 study centres in Europe, South Africa, Australia, New Zealand, and North and South America. PATIENTS AND METHODS: In total, 1550 paediatric patients with atopic dermatitis were studied. Pimecrolimus cream 1% was used twice daily to treat atopic dermatitis. The three short-term studies were placebo controlled. The two long-term studies evaluated the efficacy and safety of early intervention with pimecrolimus to prevent progression to disease flare requiring topical corticosteroid treatment, compared with reactive treatment with topical corticosteroids to treat flares of atopic dermatitis. MAIN OUTCOME MEASURES: Five parameters were measured: (i) the EASI (range of score 0-72); (ii) Investigators' Global Assessment (IGA), using a six point (0-5) scale; (iii) patients' assessment, using a four-point (0-3) scale; (iv) severity of pruritus assessment, using a four-point (0-3) scale; and (v) a quality-of-life evaluation. RESULTS: The EASI score varied in parallel and in correlation with the IGA, pruritus and patients' assessment. All correlation coefficients were statistically different from 0 (P < 0.05). The EASI correlated well with each of its components, and all paired comparisons were within agreed limits. The EASI showed good sensitivity to changes in severity. CONCLUSION: In a large, multinational patient population with atopic dermatitis, the EASI showed good validity, reliability and sensitivity to change and correlated well with other measures of severity. It therefore qualifies as a valid method of assessment in clinical studies of atopic dermatitis. PMID- 14746624 TI - The effect of environmental tobacco smoke on eczema and allergic sensitization in children. AB - BACKGROUND: The negative impact of environmental tobacco smoke (ETS) on airway diseases in children is well known. Whether there is an effect on atopic eczema is not clear. OBJECTIVES: To determine the impact of ETS on atopic eczema, allergic sensitization and allergic airway diseases in 1669 school beginners. METHODS: The prevalence of atopy-related health outcomes was assessed by questionnaire, dermatological examination, skin prick testing and specific immunoglobulin E measurement. Exposure assessments were based on measurement of cotinine [expressed as cotinine to creatine ratio (CCR)] in spot urine samples (n = 1220) together with questionnaire and interview data on smoking behaviour of the parents. RESULTS: In the total study group, prevalence of atopic eczema diagnosed on examination was significantly associated with urinary CCR values. The odds ratio (OR) and 95% confidence interval (CI), calculated for an increase of 100 ng mg-1 CCR was 1.97 (95% CI 1.23-3.16). The prevalence of skin manifestations according to questionnaire data as well as a history of asthma, wheezing, and hay fever were positively although not significantly associated with ETS exposure. When genetically predisposed children (defined by the presence of parental atopy) were compared with children whose parents had no atopy, the ORs of allergic outcome variables were generally higher in the first group. In the group of predisposed children, significant associations with urinary CCR were found for allergic sensitization against house dust mites as measured by skin prick test (OR 3.10, 95% CI 1.63-5.90). CONCLUSIONS: Children are at a higher risk of developing an atopic eczema when exposed to ETS and genetically predisposed children are at higher risk of developing a sensitization against house dust mites. PMID- 14746625 TI - Treatment of tinea imbricata: a randomized clinical trial using griseofulvin, terbinafine, itraconazole and fluconazole. AB - BACKGROUND: Griseofulvin has been the mainstay of treatment for tinea imbricata (TI) for decades; however, there have been few reports of efficacy of newer antifungals in the treatment of this condition. Many patients with TI have several obstacles to treatment due to their remote geographical locations and the primitive nature of their societies. OBJECTIVES: The aim of this study was to compare the efficacy of itraconazole, terbinafine and fluconazole with that of griseofulvin after 4 weeks of therapy. METHODS: Patients aged 12-76 years with the clinical diagnosis of TI were randomly assigned to one of four treatment groups: griseofulvin 500 mg twice daily for 4 weeks, terbinafine 250 mg daily for 4 weeks, itraconazole 200 mg twice daily for 1 week or fluconazole 200 mg once weekly for 4 weeks. Disease activity was monitored weekly. Laboratory measurements included monitoring complete blood count and liver function enzymes. Fifty-nine patients were included in the efficacy analysis: 13 in the fluconazole group, 15 in the griseofulvin group, 12 in the terbinafine group and 19 in the itraconazole group. RESULTS: Significant remission was achieved in the terbinafine and griseofulvin groups, lasting up to 8 weeks after cessation of therapy. The fluconazole group experienced no significant remission, and remission was of short duration in the itraconazole group. No adverse events were reported, and non-compliance with medications or follow-up was the only reason for removal from the study. CONCLUSIONS: Griseofulvin and terbinafine are effective in the treatment of TI. The decision of whether to treat at all and which medication to choose depends greatly on the extent of involvement, the social situation, and the availability of resources such as laboratory testing and follow-up. PMID- 14746626 TI - Clinical effectiveness of a silk fabric in the treatment of atopic dermatitis. AB - BACKGROUND: In children with atopic dermatitis (AD), eczema is easily aggravated by contact with irritant factors (e.g. aggressive detergents, synthetic and woollen clothes, climatic factors). OBJECTIVES: To evaluate the effectiveness of a special silk fabric (MICROAIR DermaSilk) in the treatment of young children affected by AD with acute lesions at the time of examination. METHODS: Forty-six children (mean age 2 years) affected by AD in an acute phase were recruited: 31 received special silk clothes (group A) which they were instructed to wear for a week; the other 15 served as a control group (group B) and wore cotton clothing. Topical moisturizing creams or emulsions were the only topical treatment prescribed in both groups. The overall severity of the disease was evaluated using the SCORAD index. In addition, the local score of an area covered by the silk clothes was compared with the local score of an uncovered area in the same child. All patients were evaluated at baseline and 7 days after the initial examination. RESULTS: At the end of the study a significant decrease in AD severity was observed in the children of group A (mean SCORAD decrease from 43 to 30; P = 0.003). At the same time, the improvement in the mean local score of the covered area (from 32 to 18.6; P = 0.001) was significantly greater than that of the uncovered area (from 31 to 26; P = 0.112). CONCLUSIONS: The use of special silk clothes may be useful in the management of AD in children. PMID- 14746627 TI - Plasma trough levels of mycophenolic acid do not correlate with efficacy and safety of mycophenolate mofetil in psoriasis. AB - BACKGROUND: Mycophenolate mofetil (MMF) has been shown to be effective for systemic treatment of psoriasis. MMF is the prodrug of mycophenolic acid (MPA), the pharmacologically active compound. The measurement of plasma MPA levels could be useful for optimizing therapeutic management using MMF. OBJECTIVES: To investigate whether plasma trough levels of MPA correlate with the efficacy and safety of oral MMF in the treatment of patients with psoriasis. METHODS: Six patients (four women and two men, mean age 58 years) with severe chronic plaque type psoriasis were treated with oral MMF 1 g twice daily. The Psoriasis Area and Severity Index (PASI), routine laboratory examinations and plasma MPA trough levels, measured by an enzyme-multiplied immunoassay (EMIT), were determined at 2 weeks and 1, 3, 5 and 7 months. RESULTS: All the patients experienced a marked improvement within the first 15 days and continued to do so for 5-7 months. Two patients achieved complete remission. MMF was well tolerated. MPA levels showed a wide intra- and interindividual variability. There was no significant correlation between MPA trough levels and the reduction of the PASI or the presence of adverse effects, but a good correlation with therapeutic compliance. CONCLUSIONS: The monitoring of MPA trough levels with EMIT appears to be a poor predictor of efficacy or toxicity. In contrast, it is a useful tool to evaluate the degree of therapeutic compliance. PMID- 14746628 TI - The occurrence of type 1 diabetes in patients with dermatitis herpetiformis and their first-degree relatives. AB - BACKGROUND: The increased prevalence of type 1 diabetes (T1D) is well documented in patients with coeliac disease, whereas evidence is scanty in patients with dermatitis herpetiformis (DH). OBJECTIVES: To assess the prevalence of T1D in patients with DH and their first-degree relatives, and to study how DH patients with associated T1D respond to a gluten-free diet (GFD) treatment. METHODS: A series of 1104 consecutive patients with DH was recorded and a specific questionnaire sent to 341 of these for familial disease surveillance. Sex- and age-matched patients with isolated DH served as controls in the diet treatment analysis. RESULTS: Twenty-five (2.3%) patients with DH were affected by T1D and three (3.0%) of their first-degree relatives also were affected by T1D, the frequencies being significantly higher than in the general population. Most DH patients with T1D and with isolated DH could adhere strictly to the GFD. The response was good or moderate in 84% of the DH patients with T1D and in 94% of the patients with isolated DH. CONCLUSIONS: The prevalence of T1D is increased in patients with DH and their first-degree relatives. The rash in DH patients with T1D responds to a GFD in a way similar to that seen in patients with isolated DH. PMID- 14746629 TI - Acute generalized exanthematous pustulosis associated with pseudoephedrine. AB - Acute generalized exanthematous pustulosis (AGEP) is an uncommon skin disorder most often caused by drugs. Few adverse reactions to sympathomimetic drugs have been reported, despite their extensive use. Although the aetiology of AGEP remains uncertain, recent data have reported involvement of drug-specific T cells and interleukin (IL)-8 production. We characterized an adverse reaction to pseudoephedrine both clinically and immunologically. Histological analysis of skin biopsies confirmed the clinical entity as AGEP, while epicutaneous tests confirmed the specificity of the reaction to the drug. Moreover, immunohistochemical studies showed a mononuclear infiltrate consisting of activated memory T cells in addition to polymorphonuclear cells. Reverse transcription-polymerase chain reaction revealed an increased expression of IL-8 in AGEP-affected skin. PMID- 14746630 TI - Allergic contact dermatitis to 5-aminolaevulinic acid methylester but not to 5 aminolaevulinic acid after photodynamic therapy. AB - We report a patient with an allergy induced by photodynamic therapy (PDT) following simultaneous treatment with both 5-aminolaevulinic acid (ALA) and ALA methylester (ALA-ME). After several PDT treatments the patient presented with acute eczema of the treated areas and itch and hyper-reactivity of the untreated skin. Patch testing demonstrated a strong +++ reaction to ALA-ME only, indicating that derivatives common to ALA and ALA-ME were not involved. This is the first case of allergy to ALA-ME. PMID- 14746631 TI - Infliximab therapy for sarcoidosis (lupus pernio). AB - Sarcoidosis is a chronic granulomatous disease of unknown aetiology in which the primary cytokine tumour necrosis factor (TNF)-alpha appears to play a major role. Older immune-modulating drugs including corticosteroids, antimalarials and thalidomide, as well as cytotoxic drugs with immune modulatory effects, have been used to control disease. We present a patient with severe mutilating cutaneous sarcoidosis (lupus pernio) who had showed only partial response to courses of a wide spectrum of immune modulators and cytotoxic therapies, and who had developed significant side-effects due to prolonged high-dose corticosteroids. However, the patient's cutaneous disease responded rapidly to the TNF-alpha inhibitor infliximab. PMID- 14746632 TI - Capillary leak syndrome induced by acitretin. AB - The mucocutaneous side-effects of systemic retinoids are well known. We report a patient with diffuse oedema, which is a very rare side-effect of retinoids. The pathophysiology of this condition is poorly known because of the rarity of relevant observations, but apparently corresponds to capillary leak syndrome. The outcome is invariably favourable following withdrawal of the drug, which should be definitively contraindicated in the patient. PMID- 14746633 TI - Annular leucocytoclastic vasculitis induced by chlorzoxazone. PMID- 14746634 TI - Infectious and toxic cellulitis due to suicide attempt by subcutaneous injection of ricin. PMID- 14746635 TI - Influence of environmental temperature on the occurrence of non-necrotizing cellulitis of the leg. PMID- 14746636 TI - Malignant proliferating trichilemmal tumour and CAV (cisplatin, adriamycin, vindesine) treatment. PMID- 14746637 TI - Efficacy of danazol treatment in a patient with the new variant of hereditary angio-oedema (HAE III). PMID- 14746638 TI - Lisinopril and Kaposi's sarcoma. PMID- 14746640 TI - Serum concentrations of vascular endothelial growth factor-D in angiosarcoma patients. PMID- 14746641 TI - Verruciform xanthoma of the scrotum in a renal transplant patient. PMID- 14746642 TI - Control of childhood pemphigus erythematosus with steroids and azathioprine. PMID- 14746643 TI - Reduced therapeutic activity of warfarin during treatment with oral isotretinoin. PMID- 14746644 TI - Delayed onset of bullous reaction with severe deep skin necrosis in association with sertraline. PMID- 14746645 TI - Acral ulcerations and osteolysis, a severe form of the carpal tunnel syndrome. PMID- 14746646 TI - Scleromyxoedema-like eruption following haemodialysis or nephrogenic fibrosing dermopathy? PMID- 14746647 TI - Lichen sclerosus-lichen planus overlap in a patient with hepatitis C virus infection. PMID- 14746648 TI - A peculiar type of dystrophic calcinosis cutis affecting the surface of damaged genital skin and mucosa in an elderly woman with a vesicovaginal fistula. PMID- 14746649 TI - Generalized eruptive histiocytosis: a possible therapeutic cure? PMID- 14746650 TI - A case of reticular erythematous mucinosis treated with topical tacrolimus. PMID- 14746651 TI - A CD56-negative case of blastic natural killer-cell lymphoma (agranular CD4+/CD56+ haematodermic neoplasm). PMID- 14746653 TI - A mathematical model for electrical stimulation of a monolayer of cardiac cells. AB - BACKGROUND: The goal of our study is to examine the effect of stimulating a two dimensional sheet of myocardial cells. We assume that the stimulating electrode is located in a bath perfusing the tissue. METHODS: An equation governing the transmembrane potential, based on the continuity equation and Ohm's law, is solved numerically using a finite difference technique. RESULTS: The sheet is depolarized under the stimulating electrode and is hyperpolarized on each side of the electrode along the fiber axis. CONCLUSIONS: The results are similar to those obtained previously by Sepulveda et al. (Biophys J, 55: 987-999, 1989) for stimulation of a two-dimensional sheet of tissue with no perfusing bath present. PMID- 14746654 TI - The experts speak. Changing landscape of opportunistic infections. Interview by Vicki Glaser. PMID- 14746655 TI - Treatment of Mycobacterium avium complex immune reconstitution disease in HIV-1 infected individuals. AB - Immune reconstitution disease caused by Mycobacterium avium complex (MAC) infection presenting shortly after the introduction of highly active antiretroviral therapy (HAART) has been reported with increasing frequency in persons with HIV-1 infection during the past several years. Several therapeutic modalities have been utilized for this entity, but the optimal means of treating MAC immune reconstitution disease remains unclear. We now describe a patient who underwent some of these therapies. We then review the therapeutic outcomes from the numerous case reports of this disorder. Finally, we propose recommendations and a clinical algorithm regarding the optimal means of treatment of MAC immune reconstitution disease during HIV-1 infection. PMID- 14746656 TI - HIV and hepatitis B: a review. PMID- 14746657 TI - Psychiatric and social barriers to HIV medication adherence in a triply diagnosed methadone population. AB - Although antiretrovirals can prolong life, medication adherence also poses a constant challenge for HIV-infected individuals because the success of antiretroviral regimens demands nearly perfect adherence to medications. This paper describes the psychiatric and social barriers to adherence in a convenience sample of HIV-positive clients in methadone treatment in the Bronx, New York. The study sample was part of a national study of HIV treatment adherence and health care utilization among triply diagnosed populations, the HIV/AIDS Treatment Adherence Health Outcomes and Cost Study. The triply diagnosed study sample is defined here as HIV-infected individuals who screened into the study with at least one psychiatric diagnosis in addition to opioid dependence on agonist therapy (methadone treatment) and at least one substance use diagnosis. Interviewers utilized modified versions of the Structured Clinical Interview for DSM-IV Disorders (SCID-I), the Structured Clinical Interview for DSM-IV Axis II Personality Disorders (SCID-II), and the Addiction Severity Index (ASI-Lite), among a battery of Cost Study instruments. Results showed that within this sample, borderline personality disorder was significantly associated with nonadherence to HIV medications. A related finding showed a significant relationship between serious social/family problems and nonadherence. These findings build on previous research on the impact of psychiatric illness on HIV medication adherence and suggest that psychiatric assessment and treatment options be linked to adherence interventions. PMID- 14746659 TI - Symptoms of posttraumatic stress and death anxiety in persons with HIV and medication adherence difficulties. AB - The present study examined the frequency and correlates of self-reported symptoms of posttraumatic stress among patients with HIV and medication adherence problems. Self-report data revealed that more than half of participants met diagnostic criteria for posttraumatic stress disorder (PTSD) according to the Posttraumatic Diagnostic Scale PDS. Death anxiety was associated with overall PTSD symptom severity scores as well as severity scores for reexperiencing, avoidance, and arousal symptoms. The association between death anxiety and total PTSD severity, reexperiencing and avoidance symptom scores remained after controlling for depression and satisfaction with social support. Anxiety, as manifested by PTSD symptoms should be routinely assessed among patients with HIV, and variables such as death anxiety, social support, and depression are appropriate targets for mental health interventions. PMID- 14746658 TI - Theory-guided, empirically supported avenues for intervention on HIV medication nonadherence: findings from the Healthy Living Project. AB - OBJECTIVES: Adherence to antiretroviral therapy (ART) remains a challenge in efforts to maximize HIV treatment benefits. Previous studies of antiretroviral adherence are limited by low statistical power, homogeneous samples, and biased assessment methods. Based on Social Action Theory and using a large, diverse sample of men and women living with HIV, the objectives of the current study are to clarify correlates of nonadherence to ART and to provide theory-guided, empirically supported direction for intervening on ART nonadherence. DESIGN: Cross-sectional interview study utilizing a computerized interview. SETTING: Recruited from clinics, agencies, and via media ads in four U.S. cities from June 2000 to January 2002. PARTICIPANTS: Two thousand seven hundred and sixty-five HIV positive adults taking ART. MAIN OUTCOME MEASURE: Computer-assessed self-reported antiretroviral adherence. RESULTS: Thirty-two percent reported less than 90% adherence to ART in the prior 3 days. A number of factors were related to nonadherence in univariate analysis. Multivariate analyses identified that being African American, being in a primary relationship, and a history of injection drug use or homelessness in the past year were associated with greater likelihood of nonadherence. Furthermore, adherence self-efficacy, and being able to manage side effects and fit medications into daily routines were protective against nonadherence. Being tired of taking medications was associated with poorer adherence whereas a belief that nonadherence can make the virus stronger was associated with better adherence. CONCLUSIONS: Results support the need for multifocused interventions to improve medication adherence that address logistical barriers, substance use, attitudes and expectancies, as well as skills building and self-efficacy enhancement. Further exploration of issues related to adherence for African Americans and men in primary relationships is warranted. PMID- 14746660 TI - Resistance seen in newly diagnosed patients. PMID- 14746661 TI - Rise in syphilis for San Francisco. PMID- 14746662 TI - Mastitis due to Mycobacterium avium complex in an HIV-infected woman taking highly active antiretroviral therapy. AB - Although infection with Mycobacterium avium complex (MAC) is common in HIV infected patients with CD4+ T cell counts less than 100/mm3, localized infection of the breast is uncommon. A brief literature review of mastitis from atypical mycobacteria is presented, along with the first reported case of localized MAC infection of the nonaugmented breast in an HIV-infected woman taking highly active antiretroviral therapy. PMID- 14746663 TI - New anti-HIV protease inhibitors provide more treatment options. AB - For several years, protease inhibitor (PI)-containing antiretroviral treatment (ART) regimens have demonstrated long-term virologic and immunologic benefits and good durability of response. However, first-generation PIs have been associated with high pill burdens, gastrointestinal side effects, perturbation of lipid levels and glucose metabolism, and, in some cases, food and hydration requirements. Coadministration of low-dose ritonavir with PIs has enhanced their pharmacokinetic profile (lower doses, fewer pills, less frequent dosing schedules) and pharmacodynamics (increased potency, especially against resistant viruses) but has also been associated with increases in lipid levels. Two new PIs, atazanavir and 908 (fosamprenavir), may offer salvageable PI treatment options and may also address issues of potency, tolerability, and convenience by requiring fewer pills and causing fewer lipid and glucose perturbations than current PI options. The availability of these novel PIs may improve longterm treatment options for many patients. PMID- 14746664 TI - Hospitalized HIV-infected patients in the HAART era: a view from the inner city. AB - To evaluate hospitalizations of HIV-infected patients in the highly active antiretroviral therapy (HAART) era, we analyzed 2736 admissions of 1562 HIV infected patients to Cook County Hospital from September 20, 1999 to July 10, 2002. Patients were predominantly African American (81%), male (72%), and active substance abusers (74%). Only 48% of patients with a prior HIV diagnosis were taking HAART and 37% of them had a viral load less than 1000 copies per milliliter. Patients on protease inhibitor (PI)-sparing regimens more frequently achieved a viral load less than 1000 copies per milliliter than those on a PI containing regimens (41% vs. 34% p = 0.036). For patients with CD4 cell counts less than 200 cells per milliliter, those not taking HAART were more likely African American (83% vs. 76%, p < 0.031), homeless (13% vs. 5%, p < 0.001), active substance abusers (79% vs. 65%, p < 0.001), female (28% vs. 22%, p = 0.001), new to the hospital system (19% vs. 6%, p < 0.001), or not recently seen in the outpatient clinic (42% vs. 17%, p < 0.001). In our population, active substance abuse was prevalent and only a minority of patients was taking HAART. Women were receiving HAART less often, independent of race and substance abuse. Aggressive programs are needed in high-risk populations to address substance abuse issues and to improve patient use of HAART. PMID- 14746665 TI - Attitudes toward HIV protease inhibitors and medication adherence in an inner city HIV population. AB - The objective of this pilot study was to examine attitudes toward protease inhibitors (PIs) among HIV-infected individuals and to assess the relationship between PI attitudes and adherence to PIs. Respondents were recruited from four AIDS service organizations in New York City; the total sample consisted of 97 HIV infected individuals who were taking a PI. The sample consisted largely of African Americans and Latinos from inner city areas, and most had a low level of education. Adherence was suboptimal, with more than 50% of respondents failing to take their PI medications exactly as prescribed within the previous month. Individuals who had recently used illegal drugs within the past 6 months were more likely to be nonadherent to PIs. Those who were not adherent to PI medications reported greater concern about the side effects of PIs and were more likely to believe that it was acceptable to skip doses of PIs. PMID- 14746666 TI - Program to enhance health literacy and treatment adherence in low-income HIV infected Latino men and women. AB - This paper reports the initial results of a pilot study to evaluate the acceptability and effectiveness of a program to enhance health literacy in low income HIV-infected Latino men and women receiving antiretroviral therapy. Participants rated the program highly on measures of satisfaction, providing evidence of its acceptability. The effectiveness of the program was assessed in comparisons of the intervention (n = 41) and standard care only (n = 40) groups at baseline and 6-week intervals. Program participants showed significant improvement over comparison group participants on measures of HIV/AIDS and treatment-related knowledge and recognition and understanding of HIV terms. Although there were no significant changes in adherence mastery and behaviors during the 6-week follow up period, there were significant changes in program participants' knowledge about medication adherence. Future steps to examine the sustainability of the program in the medical management of patients are planned in addition to determining its long-range relative impact. PMID- 14746667 TI - Syphilis and HIV prevalence among commercial sex workers in Central Java, Indonesia: risk-taking behavior and attitudes that may potentiate a wider epidemic. AB - A cross-sectional study was conducted on 200 commercial sex workers (CSWs) from two brothel communities in Central Java, Indonesia, to determine the seroprevalence of syphilis and HIV and characterize associated knowledge, beliefs, and risk-taking behaviors. A questionnaire was administered and blood drawn for HIV and syphilis serologies. Focus groups with a total of 20 women were also conducted at both communities to supplement survey data. The mean CSW age was 27.3 years; mean number of clients seen per day was 2.27. The prevalence of syphilis and HIV were 7.5% and 0.5%, respectively. Thirty percent said they and their partners never used condoms during sex, and only 3.0% said they always used condoms. The most common client groups were truck drivers and sailors. While Central Java appears to remain in a pre-epidemic state, there is enormous potential for a significant increase in HIV and STD transmission. PMID- 14746668 TI - Smoking and avascular necrosis. PMID- 14746669 TI - Early gestation viral load and HIV transmission. PMID- 14746670 TI - [New stage of child health care development in China]. PMID- 14746671 TI - [Effects of different feeding methods on the iodine status of the infants during the weaning period]. AB - OBJECTIVE: To investigate the iodine levels of urine from 1 month old breast-fed infants and the ones of milk and urine from the lactating women, and to observe the effects of different feeding methods (breast-feeding, mixed-feeding, bottle feeding) on the iodine status of the infants during the weaning period in Beijing. METHODS: From March, 2001 to March, 2002, the iodine levels of urine from 97 breast-fed infants 1 month of age and the ones in milk and urine from lactating women were measured and compared. The infants followed up were divided into 3 groups (breast-fed, mixed and bottle-fed) until 6 months old. Their iodine levels of urine were measured and compared with the ones of 1 month of age. RESULTS: The median value of urine iodine from breast-fed 1 month old infants was 183 micro g/L, suggesting that the infants with breast-fed had good iodine nutritional status. The median value of urine iodine from lactating women was 122 micro g/L, significantly lower than the value of milk iodine, 201 micro g/L (P < 0.001). which suggests that the lactating women were iodine deficient but could provide infants iodine adequately through breast feeding. Compared with 1 month af age, the urine iodine levels of 6 months old infants with breast-feeding increased (P < 0.001), the ones with bottle-feeding decreased significantly (P < 0.001) and the mixed-feeding group did not change (P > 0.05). The differences among 3 groups were significant (P < 0.005), the urine iodine levels of infants of both breast-feeding and mixed-feeding groups were higher than the ones of bottle-feeding. The breast-feeding group was the highest one among three groups. CONCLUSION: The breast-fed infants were nourished with iodine, but the lactating women were iodine deficient. Accompanied the decrease of the amount of breast milk, the iodine levels of infants urine decreased during the weaning period, some bottle-feeding infants were iodine deficient. PMID- 14746672 TI - [Blood leptin level of 154 Chinese children 0 - 14 years of age]. AB - OBJECTIVE: To study blood Leptin level of 154 (78 male, 76 female) Chinese obese/non-obese children aged 0 - 14 years during 1999 - 2001. METHODS: The gender- and age-specific distribution pattern of Leptin and its relationship with anthropometric parameters (waist circumference, waist/hip ratio, lean body mass, fat mass, body fat percentage, BMI/Kaup index etc.) and blood insulin level were recorded. RESULTS: (1) The blood Leptin level in healthy non-obese kids ranged from 1.01 - 29.92 (ng/ml), the mean values and SD were 2.99 +/- 2.13 (ng/ml) [90% confidence interval was 1.36 - 14.21 (ng/ml) in boys and 1.74 - 21.17 (ng/ml) in girls]. There was no significant difference in the blood Leptin level between serum and plasma. (2) The blood Leptin level was higher in overweight/obese kids than that in non-obese kids (P < 0.001). (3) There was significant difference in the blood Leptin levels between boys and girls groups (P = 0.023), especially in non-obese group (P = 0.004). The multiple regression analysis showed that there was no correlation between gender and blood Leptin level when body fat factor was added (P = 0.138, 0.241, 0.990), but there was still a strong correlation between blood leptin level and BMI, FM and BF% (P < 0.001). (4) There was a correlation between blood Leptin level and age (P = 0.005), especially in overweight/obese group and in girls (P = 0.001). The blood Leptin level rose from early puberty, especially in girl group (P = 0.045). There was significant difference in blood Leptin level in different age groups (P < 0.001) (5) There were strong positive correlation between blood Leptin level and BMI, BM and FM%, a weak correlation with LBM, and no correlation with W/H ratio in boys and a positive relationship in girls. The Quatatic equation was better than the linear equation in description of the correlation mentioned above. (6) There was a correlation between blood Leptin from 0 to 7 yr and birth weight (P = 0.001), after 7 yr of age this correlation disappeared (P = 0.456). (7) A positive correlation was seen between blood Leptin level and blood insulin level (P < 0.001). CONCLUSION: The blood Leptin level of 0 - 14 years old children is consistent with the level of growth of adiposity tissue and the degree of adiposity rebound. PMID- 14746673 TI - [Association of the vitamin D receptor gene start codon polymorphism with vitamin D deficiency rickets]. AB - OBJECTIVE: Vitamin D deficiency rickets often causes growth retardation, impaired bone formation and hypocalcemia in children. It is well known that rickets is mainly caused by vitamin D deficiency, but whether there is hereditary susceptibility of children to develop vitamin D deficiency rickets is unknown. Vitamin D receptor (VDR) gene has been used as one of genetic markers in studying the metabolic diseases of bone. The present study aimed to explore the hereditary susceptibility of children to develop rickets through studying the association between VDR gene start codon polymorphism and vitamin D deficiency rickets, METHODS: The subjects were selected from Kunming city, every subject was of Han ethnic group. The subjects were composed of two groups, the patient group consisted of 48 children with active vitamin D deficiency rickets which was diagnosed clinically and confirmed radiologically; the control group was composed of 92 normal children. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), DNA sequence analysis and genetic analysis methods were used. A restriction fragment length polymorphism in the start codon of VDR gene (FokI) was tested in both groups. RESULTS: VDR gene start codon polymorphism was tested successfully for every subject. Frequencies of FF, Ff and ff genotypes were 46%, 33% and 21% in the rickets group, and 22%, 52% and 26% in the control group, respectively. A significant difference was found in the frequency distribution of VDR genotype between two groups (chi(2) = 8.912, P = 0.012). In the patient group, Ff and ff genotypes were less common than control group, but the FF genotype was more common than control group (OR = 3.046), indicating that FF genotype may be significantly associated with vitamin D deficiency rickets. Moreover, VDR allele frequencies of FokI polymorphism also showed significant difference between the two groups (chi(2) = 5.451, P = 0.020), F alleles were more common in patient group than in control group. DNA sequence analysis identified that the start codon of F allele was mutated from ATG to ACG. CONCLUSION: There is an association between VDR gene start codon polymorphism and vitamin D deficiency rickets. This study suggested the possibility that VDR gene polymorphism might be important in determining an individual's susceptibility to development of vitamin D deficiency rickets. PMID- 14746674 TI - [Evaluation on intervening efficacy of health education on accidental suffocation and drowning of children aged 0 - 4 in countryside]. AB - OBJECTIVE: The main causes of death in children aged 0 - 4 were accidental suffocation and drowning which had a significant relation with parents' lack of prevention knowledge and effective measures. By comparing parents' acknowledge and behavior in preventing accidental suffocation and drowning and the mortality rate of accidental suffocation and drowning after intervening with those before the intervening, evaluation was made on intervening efficacy of health education. METHODS: Six counties in north Jiangsu were involved in the survey. Two townships drawn from each county were divided into the intervened group and the controlled group. Health education was carried out in the intervened group for one year in 2000. Twenty to 30 parents of infants and children aged 1 - 4 drawn randomly from every township in the intervened group were taken as investigation subjects before and after intervening, and a questionnaire was made for them to answer. Meanwhile, the mortality rates of accidental suffocation and drowning were measured. RESULTS: Followed by health education for a year, the parents' opinion about whether unintentional injury can be avoided or not has changed from 18.8% to 20.5% to 1.8% - 2.9%, and parents' knowledge about how to give first aid in spot has increased from 11.1% - 13.5% to 41.1% - 56.8%. The parents' behavior that not sleeping with their infants in the same beds and not tying infants in a candle with blanket, and setting up fence beside pools and rivers has increased by 75.7%, 61.5% and 61.2%, respectively, while their relative knowledge has increased by 212.7% and 194.3%. In the intervened group, the mortality rates of infants' accidental suffocation per 100,000 has fallen from 487.8 to 71.2, dropped by 85.4%; and the mortality rates of drowning in children aged 1 - 4 per 100,000 has fallen from 60.0 to 36.2, dropped by 39.7%. In comparison, in the controlled group, the mortality rates of infants' accidental suffocation per 100,000 has fallen from 344.1 to 276.4, dropped by 19.7%; and the mortality rates of drowning in children aged 1 - 4 per 100,000 has increased by 26.3%, from 51.7 to 65.3. CONCLUSION: Health education to parents is an effective intervening measure for prevention of accidental suffocation and drowning. The goal of health education should be to change inadequate behavior and dangerous environment in which unintentional injury is easily happened. The intervening measures that not sleeping with their infants in the same beds and not tying infants in a candle with blanket, and putting up fence beside pools and rivers are feasible and practicable. PMID- 14746675 TI - [Investigation and analysis of child neglect in 1163 urban children aged 3 - 6 years]. AB - OBJECTIVE: Child abuse and neglect (CNS) increasingly have become a serious problem in modern society all over the world. Children neglected and abused are at risk for maladaptation in behavioral, emotional, social, cognitive, and physical functioning. The purpose of this investigation was to develop the Child Neglect Scale for children aged 3 - 6 years in Chinese urban areas and to test the reliability and validity of this norm. METHODS: According to the principle and procedure of developing scales, a child neglect questionnaire was designed. Multistage sampling was used in the study in accordance with sex, age, the levels of economy and kindergartens, a random sample of 1,465 children aged 2 - 6 years was selected from 25 cities. Of the 1,465 subjects who completed the Child Neglect Questionnaires again 2 weeks later, 66 had completed data on each of the two occasions. The Child Neglect Questionnaires were completed by parents and caregivers. The data were analyzed using items analysis, factor analysis, internal consistency analysis and the reliability and validity of the norm were tested. The factor structure of the Child Neglect Scale was determined using an oblique rotation. The internal consistency was reflected in Cronbach's alpha. Since no existing categorization scheme on child neglect could be found in the literature, one was formulated in an attempt to efficiently and meaningfully group similar types of caregiver behaviors. The initial pool of potentially neglecting caregiver behaviors was drawn from literature. The most common patterns of parental behaviors were grouped according to six neglect subscale schemes found in the literature, with modifications made to increase item specificity and reduce overlap. The initial Child Neglect Questionnaire contained a 25-item physical neglect subscale, a 59-item emotional neglect subscale, a 23 item educational neglect subscale, a 20-item safety neglect subscale, a 10-item medical neglect subscale, a 9-item social neglect subscale. RESULTS: Completed Child Neglect Questionnaires were obtained from 1,457 subjects, 726 males and 731 females. The overall findings of this study came from 1,163 subjects, because children aged 2 years (294 subjects) were removed from subjects by statistical method. The 91-item Child Neglect Scale was presented to the respondent as a child neglect questionnaire with a 9-item social neglect subscale and some items in other subscales were removed from the initial questionnaire by using statistical method. The questionnaire contained 17 physical neglect items, 40 emotional neglect items, 17 educational neglect items, a 10 safety neglect items, and 7 medical neglect items. The internal consistency of the overall Child Neglect Scale as reflected in Cronbach's alpha was 0.94, the split-half reliability was 0.88, the test-retest reliability was 0.92. For physical, emotional, educational, safety, medical neglect subscale, Cronbach's alpha, respectively was 0.80, 0.88, 0.84, 0.68, 0.60, the split-half reliability was 0.54 - 0.88, the test reliability was 0.89, 0.94, 0.87, 0.80, 0.77, respectively. The construct validity, face validity and exterior validity of the norm were good. For physical, emotional, educational, safety, medical neglect subscale and overall child neglect scale, the scores of percentiles 90 were used as cut-point. They were 35, 80, 40, 20, 15, 190, respectively. CONCLUSIONS: The norm was consistent with China's economic-cultural background and reality. It not only may be used for research purpose but also may be useful in assessing parent's and child-oriented individual's behaviors and attitude as a measuring instrument. PMID- 14746676 TI - [Influences of environmental endocrine disruptors on growth, development and health of children]. PMID- 14746677 TI - [Metallothionein and prevention and treatment of environmental risk factors for health]. PMID- 14746678 TI - [Results and suggestion from comparing between world excellent protocols of childhood acute lymphoblastic leukemia]. PMID- 14746679 TI - [Clinical analysis of 29 children with early infectious complications following hematopoietic stem cells transplantation]. AB - OBJECTIVE: To study the clinical features and the incidence of early infectious complications following children hematopoietic stem cells transplantation (HSCT). METHODS: The clinical data of 29 cases with early infectious complications following HSCT was retrospectively analyzed. RESULTS: The incidence of early infectious complications following HSCT in 31 children (including 22 cord blood transplantation and 9 peripheral blood stem cells transplantation) was 94% (29/31). The first occurrence of the early infectious complications was at a median of 6 (0 - 22) days, the peak time of incidence was at a median of 4 - 7 days post transplantation. The duration of the first early infectious complications was at a median of 9 (3 - 20) days. The occurrence of the second early infectious complications was at a median of 19 (13 - 27) days. For all of the 29 children, when they developed early infectious complications their absolute neutrophil counts (ANC) were all > 0.5 x 10(9)/L. The most common infectious sites were the digestive tract (oral and gastro-intestinal mucositis) and then the respiratory tract. Gram negative blood infections were quite frequent and Pseudomonas aeruginosa was common in the oral-pharynx discharge cultures. Two children had Mycoplasma pneumonia infections and there were 4 incidences with fever but no definite infectious foci. The incidence and duration of early infectious complications following hematopoietic stem cells transplantation were associated with the duration of neutropenia. The source and the MNCs dose of the graft, the difference of conditioning regimen and GVHD prophylaxis method did not have a significant impact on the incidence and duration of early infectious complications. Antibiotic prophylaxis (including Tienam) could delay the occurrence of the early infections significantly. CONCLUSION: The incidence and duration of early infectious complications following hematopoietic stem cells transplantation were directly associated with the duration of neutropenia. Tienam regimen could postpone the early infections incidence and had effect of preventing the early infectious complications. PMID- 14746680 TI - [Reversal of multidrug resistance in leukemic cell line K562/AO2 by chlordelazine in vitro]. AB - OBJECTIVE: Some recent studies revealed that phenthiazine might be able to reverse tumor cell drug-resistance. Chlorderazin belongs to the phenthiazine compounds. The study aimed to investigate the reversing effect and mechanism of chlorderazin on multidrug resistance of leukemic cell line K562/AO2. METHODS: (1) The cytotoxicities of chlorderazin were assayed with the tetrazolium dye, 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method. (2) The reverse effect of chlorderazin on K562/AO2 cells was analyzed with MTT method. The multidrug resistance reversal index (RI) was equal to the ratio of control group IC(50)/test group half inhibition concentration IC(50). (3) The intracellular daunorubicin (DNR) concentrations were measured by the flow cytometry. (4) Mdr1 mRNA expression was detected by reverse transcription polymerase chain reaction (RT-PCR). The ratio of mdr-1/beta-actin density was calculated. RESULTS: (1) Chlorderazin 3 micro g/ml showed little toxicity to K562/AO2 cells and the suppression rate was less than 5%, so the concentration of 3 micro g/ml chlorderazin was selected as the experiment concentration. (2) The cytotoxicities of DNR to K562/AO2 were enhanced by 3 micro g/ml of chlorderazin (P < 0.05) and RI was 1.901. (3) Chlorderazin of 3 micro g/ml could increase the intracellular DNR accumulation significantly (P < 0.05), and the fluorescence staining by the flow cytometry was higher (250.95 +/- 18.96) than the control group (112.75 +/- 15.78) and shift right in K562/AO2 cells treated with chlorderazin, and the difference was significant (P < 0.05). (4) Chlorderazin has no significant influence to the expression level of mdr-1 mRNA. Both test group and control group showed a clear mdr-1 mRNA band located at the position of 157 kb. The ratios of mdr-1/beta-actin density were 0.414 +/- 0.012 in the test group and 0.447 +/- 0.027 in the control group, respectively, and the difference was not significant (P > 0.05). CONCLUSION: Chlorderazin could reverse the multidrug resistance by increasing the intracellular DNR accumulation in K562/AO2 cells. The effects had no correlation to the mdr-1 gene. Further study is needed. PMID- 14746682 TI - [Transcatheter closure of atrial septal defects in 40 pediatric patients]. AB - OBJECTIVE: To evaluate the clinical efficiency of transcatheter closure of atrial septal defect (ASD) with AGA-Amplatzer occlusion device in pediatric patients. METHODS: Forty patients with ASD, 16 males, 24 females, at a mean age of 10.2 years (ranged from 3 to 15 years of age) and with a mean weight of 35.8 kg (ranged from 11 to 87 kg) were studied. Six cases were complicated with pulmonary stenosis (PS), 1 was complicated with ventricular tachycardia (VT). Right heart catheterizations were done in 40 patients for measuring the pressures of right ventricle and pulmonary artery. The balloon diameter of ASD was measured using balloon catheter with guiding wire. The diameter of ASD was measured by TTE and/or TEE, ascertaining the location and size of ASD. Amplatzer occlusion device was sized to be equal to or 1 - 2 mm more than the diameter of balloon stretched. RESULTS: All patients had successful implantation of the Amplatzer device. The success rate was 100%. The diameter measured by TTE was 7 - 30 mm (mean 17.12 mm). The diameter measured by TEE was 7 - 32 mm (mean 18.44 mm). The diameter of balloon stretched of ASD was 8 - 34 mm. Of the 40 cases, 6 were complicated with PS and accepted percutaneous balloon valvuloplasty (PBPV). One case was complicated with VT and accepted radiofrequency catheter ablation (RFCA). Neither complication nor residual shunt was found in any of the patients. The patients were recovered and followed up for 3 or 4 days after deployment of the Amplatzer device. Clinical symptom, cardiac murmur, and findings in ECG, echocardiography and X-ray were improved markedly. CONCLUSION: AGA-Amplatzer occlusion device is safe and efficient in pediatric patients with ASD. PMID- 14746681 TI - [Erythropoietin increases transferrin receptor expression and the impact of erythropoietin on K562 leukemic cell cycle]. AB - OBJECTIVE: Functionally, erythropoietin (EPO) can promote the proliferation and growth of erythroid progenitor cells, and it is widely used in the treatment of anemia in chronic diseases caused by tumor and inflammation. However, it is unclear whether EPO has any effect on tumor cell iron metabolism and tumor cell proliferation. The purpose of this study was to explore the effects of recombinant human EPO (rhEPO) on the expression of transferrin receptor (TfR, CD(71) antigen) of leukemic cell K562 and its relation to cell cycle. METHODS: In vitro culture of K562 cell was performed with additions of various concentrations of rhEPO and Fe. Treatments were terminated at 24 h and 72 h, respectively. Then each group of cells was incubated with FITC-IgG antibody to CD(71) or PI, a kind of DNA dye. And TfR expression and DNA synthesis status were analyzed by flow cytometry. RESULTS: (1) The expression of TfR by K562 cells increased significantly when incubated for 72 h with different concentrations of rhEPO. The measurement values of 5 U/ml, 10 U/ml and 20 U/ml groups were 12.2 +/- 1.40, 10.7 +/- 0.99 and 11.1 +/- 0.90, respectively. They were markedly increased when compared with that of control group (6.27 +/- 0.11, P < 0.05). (2) When incubated with rhEPO (5 u/ml) alone or combined with FeCl(3) (100 micro mol/L), the percentages of cells in S phase were 51.1% and 59.6%, respectively. They significantly increased when compared with that of control group (42.9%, P < 0.05). CONCLUSIONS: Iron is very important for the proliferation of both normal cells and leukemic cells. It is essential to the activity of ribonucleotide reductase (RR). The authors hypothesized that rhEPO would increase the expression of TfR and intracellular iron content of leukemic cells, which would enhance the DNA synthesis and cell proliferation. Therefore, the clinical application of rhEPO to promote erythropoiesis of cancer patients should be cautious. PMID- 14746683 TI - [Change of transforming growth factor beta in peripheral blood mononuclear cell of children with nephrotic syndrome and its significance]. AB - OBJECTIVE: Idiopathic nephrotic syndrome (INS) is a common glomerular disease. The pathogenesis of the disease remains unclear. Recent studies indicate that transforming growth factor beta (TGF beta) is the main cytokine involved in glomerular disease. It plays an important role in the development of INS and in occurrence of glomerulosclerosis. The present study aimed to study changes and significance of TGF beta in children with idiopathic nephrotic syndrome (INS). METHODS: Totally 35 cases with INS (13 males, 22 females) were studied. The age of onset was between 2 years and 1 months and 14 years with an average of 8 years and 3 months. The active stage group had 35 cases and the remission stage groups had 25 cases. The cases in active stage group had first onset of the disease with obvious clinical symptoms and abnormal laboratory findings without use of corticosteroids. The cases in remission stage group were asymptomatic without abnormal laboratory findings. Protein in urine was negative over 4 weeks after oral administration of prednisone for 8 weeks. Twenty five cases were steroid responsive and 10 cases were steroid non-responsive among the 35 cases. Thirty healthy young children were enrolled as control. TGF beta was detected by ELISA in peripheral blood mononuclear cell (PBMC) culture medium. The TGF beta mRNA gene expression was measured by in situ PCR in PBMC. RESULTS: (1) Concentration of TGF beta(247 +/- 26) ng/L and TGF beta mRNA expression (0.57 +/- 0.18) in active stage of simple type or nephritis type INS were higher than those of remission stage and control (P < 0.01). Concentration of TGF beta[(125 +/- 16) ng/L] and TGF beta mRNA expression (0.30 +/- 0.12) in remission stage were higher than that of control (P < 0.05). (2) The level of TGF beta protein in nephritis type [(275 +/- 26) ng/L] was significantly higher than that in simple type [(220 +/- 18) ng/L] in active stage INS (t = 6.45, P < 0.01). No significant difference in TGF beta mRNA expression was found between the nephritis type (0.58 +/- 0.15) and simple type (0.55 +/- 0.16) in active stage INS, either (P > 0.05). But these two types were different from the control (P < 0.01). (3) Concentration of TGF beta and TGF beta mRNA expression after therapy was clearly lower than that before therapy in steroid responsive group (P < 0.01). Whereas no significant change was seen in steroid non-responsive group. Both indicators were higher in steroid non-responsive group than in steroid responsive group whether before or after therapy. CONCLUSION: TGF beta may play an important role in the mechanism of INS and its level in PBMC can be used as an immunological indicator for the illness state, therefore, determination of TGF beta level and mRNA may be of some clinical significance. PMID- 14746684 TI - [Individual allergens as risk factors for asthma and bronchial hyperresponsiveness in Chinese children]. AB - OBJECTIVE This study aimed to determine the relationship between individual allergens with current wheezing and bronchial hyperresponsiveness (BHR) in schoolchildren from three chinese cities: Beijing, Guangzhou and Hong Kong. METHODS Community-based random samples of 10-yr-old schoolchildren from the 3 cities were recruited for study using the International Study of Asthma and Allergies in Childhood (ISAAC) Phase II protocol. The subjects were studied by parental questionnaires (n = 10,902), skin-prick tests (n = 3478), and methacholine challenge tests (n = 608). RESULTS The highest prevalence rates of wheezing in the past 12 months (Beijing, 3.8%; Guangzhou, 3.4%; Hong Kong, 5.8%) and atopy (Beijing, 23.9%; Guangzhou, 30.8%; Hong Kong, 41.2%, defined as having 30 mmHg after a mean period of 15.3 months post-surgery. Mean transhomograft pressure gradient was 19.8 (16.2%) (range, 2-100 mmHg). All patients were in functional class I, except 2 who were in New York Heart Association class II with severe stenosis. These 2 patients were treated percutaneously with stent placement and no reoperation. No association was found between clinical outcome and sex, age or homograft diameter. We found a trend toward greater perioperative use of plasma, platelets and red cells in the group of patients in comparison to controls, although the difference was significant only for postoperative use of plasma (1.7 [3] vs 5.5 [5.3] units; P<0.05). Cardiac magnetic resonance imaging showed stenosis predominantly in the body of the homograft, whereas the valvular component itself remained competent. Right ventricular hypertrophy was mild or very mild in 7 patients and moderate in 2. CONCLUSIONS: The incidence of some degree of pulmonary homograft dysfunction after the Ross procedure was non-negligible, but its clinical impact seems to be slight. Patients with severe stenosis were treated successfully via a percutaneous approach. The use of blood products might be a risk factor for the development of this complication. PMID- 14746716 TI - [Non-fluoroscopic electroanatomical mapping (CARTO system) in the ablation of atrial tachycardias]. AB - INTRODUCTION AND OBJECTIVES: The recent introduction of navigation systems has made substantial improvements in cardiac electrophysiological mapping. We present our experience with non-fluoroscopic electroanatomical mapping in patients with atrial tachycardias. PATIENTS AND METHOD: We studied 24 consecutive patients with atrial tachycardias (10 of whom had undergone previous radiofrequency ablation which failed). In all patients we performed electroanatomical mapping of the atria with the CARTO system, which combines electrophysiological and spatial information and allows visualization of atrial activation in a three-dimensional anatomical reconstruction of the atrial cavity. Mapping was performed during tachycardia (22 patients) or in sinus rhythm (2 patients), using a left atrial approach in 12 patients. Cooled-tip ablation was performed in 3 patients. RESULTS: Three-dimensional mapping distinguished clearly and rapidly between reentrant (9 patients) and focal mechanisms (15 patients). Radiofrequency catheter ablation was aimed at the critical isthmus of conduction (voltage maps) in patients with macroreentrant tachycardias. For focal tachycardias the catheter was re-navigated within the target area (activation maps) to the earliest focus of ectopic impulses. Acute success was obtained in 19 patients (79.2%), with early recurrence in 2 of them. Fluoroscopy time was 60 (21 min). CONCLUSIONS: Visualization of atrial activation in a three-dimensional reconstruction of the atria with the CARTO electroanatomical mapping system facilitated the integration of electrophysiological and anatomical information in patients with atrial tachycardias. This technique is potentially helpful in ensuring successful treatment of the substrate of tachycardia in this selected group of patients. PMID- 14746717 TI - [Differences in clinical profile and outcome in patients with decompensated heart failure and systolic dysfunction or preserved systolic function]. AB - OBJECTIVES: To compare the clinical characteristics and short- and long-term prognosis for chronic heart failure with left ventricular systolic dysfunction or preserved systolic function. PATIENTS AND METHOD: Three-hundred twenty-eight consecutive patients with decompensated chronic heart failure were studied prospectively. Depending on ejection fraction, participants were classified as having systolic dysfunction (group 1, ejection fraction < or = 40%,) or preserved systolic function (group 2, ejection fraction >40%). RESULTS: Systolic dysfunction was detected in 192 patients (58.5%) and preserved systolic function in 41.5%. Mean age was 62.7 (12.5 years) in group 1 and 65.2 (16.2 years) in group 2 (P=.03), with a male prevalence of 73.3% and 49.3%, respectively (P<.001). Ischemic cardiomyopathy was more frequent in group 1 (44.8% vs 25%; P<.001). Physical examination and electrocardiogram findings were similar in both groups, except for a higher proportion of patients in group 1 with a heart third sound (43.2% vs 25%; P=.001) and left bundle branch block (40.6% vs 15.4%; P<.001) and abnomal Q waves (31.3% vs 20.6%; P=.04). In-hospital mortality was similar in patients with systolic dysfunction and preserved systolic function (2.9% vs 1%; P=NS). Twenty-four-month cumulative survival was 61% for patients with systolic dysfunction and 76% for patients with preserved systolic function (log rank test P=NS). In the Cox proportional hazards model, which included age, sex, functional class, hepatomegaly, peripheral hypoperfusion, BUN, sodium level, ejection fraction > 40%, and biventricular heart failure, preserved systolic function was not associated with late mortality. The variables that were independent predictors of late mortality were peripheral hypoperfusion (OR = 3.7; P<.0001), low sodium level (OR=0.9; P=.009) and male sex (OR=1.9; P=.041). CONCLUSIONS: Decompensated chronic heart failure with preserved systolic function was more frequent in women and older patients. Patients with preserved systolic function had a lower prevalence of coronary heart disease. However, these differences had no impact on the short- and long-term prognosis. PMID- 14746718 TI - [Postmyocardial infarction cardiac rehabilitation in low risk patients. Results with a coordinated program of cardiological and primary care]. AB - INTRODUCTION AND OBJECTIVES: To assess the efficacy of cardiac rehabilitation with a mixed primary and cardiological care program in patients with low-risk myocardial infarction. PATIENTS AND METHOD: The participants in this 12-month prospective study were 153 consecutive patients with low-risk myocardial infarction (MI) referred to their primary care center for follow-up care. Of these patients, 113 were referred to a mixed primary and specialized care program that included physical exercise, cardiovascular risk control, an antismoking program, health education talks and psychological evaluation. The other 40 patients served as controls. We analyzed the results after 3 months and 1 year of follow-up. RESULTS: There were no differences between the two groups at baseline. After 1 year, improvements were seen in smoking habit (4.6% vs 15.6%; P<.05) and body mass index (26 [2] vs 29 [2]; P<.05). Dyslipidemia, glucose and blood pressure were similar in both groups after follow-up. Greater improvements in the group of patients who participated in the program were seen after 1 year in quality of life (78 [2] vs 91 [2]; P<.05), exercise capacity (10.3 [2] vs 8.4 [3]; P<<.01) and return to active employment (84.6% vs 53.3%; P<.05). CONCLUSIONS: After 1 year of follow-up, the cardiac rehabilitation program coordinated by cardiological and primary care services for low-risk post-MI patients improved quality of life, and increased exercise tolerance, active employment, and the number of participants who quit smoking. The mixed program also reduced body mass index. These results suggest the need for similar programs. PMID- 14746720 TI - [Ionic currents and ventricular fibrillation dynamics]. AB - Ventricular fibrillation is the principal immediate cause of sudden cardiac death. Yet, in contrast to other arrhythmias, ventricular fibrillation is considered to be inaccessible to pharmacologic therapy because of its characteristic and apparently never-ending disarray of electrical waves that seem to propagate chaotically throughout the ventricles. Its prevention has historically been focused on the suppression of ventricular ectopy, with the idea of eliminating potential triggers of fibrillation, which from a clinical standpoint has proven to be detrimental. During the last decade, the application of the theory of wave propagation in non-linear excitable media to the study of cardiac fibrillation has led to a dramatic increase in our understanding of its mechanisms. It is now clear that fibrillation is generated and maintained by rotors that gyrate at exceedingly high frequencies. From such rotors emanate spiral waves of excitation that propagate throughout the myocardium in very complex ways. Among the most important factors that determine rotor dynamics are the electrophysiological properties of the ventricular cells, established by their underlying transmembrane ionic currents. Thus, in recent years, studies have focused on the roles played by specific ionic mechanisms and their modulation by antiarrhythmic drugs in ventricular fibrillation dynamics. This review article summarizes the main findings of such studies, which pave the way for a better understanding of fibrillation, and for the development of new pharmacological approaches that aim to prevent rotor formation and maintenance rather than to suppress the triggering ectopic event. PMID- 14746721 TI - [Coronary fold resembling slit-like lesion]. PMID- 14746722 TI - [Direct stenting without predilatation: influence of stent diameter on the immediate results]. AB - The influence of stent diameter in a direct stenting technique was analyzed. We retrospectively identified 987 consecutive lesions in 773 patients in whom direct stenting was attempted. Lesions were divided into two groups: group 1, nominal stent diameter 2.5 mm (237 lesions) and group 2, > or =2.75 mm (n=750). Differences between groups were found in age (64.4 [10.4] vs 62.3 [11] P=.009), female sex (33.2% vs 17%; P<.0001), diabetes (44% vs 33.1%; P=.003), tortuosity (5.4% vs 2.5%; P=.034), reference diameter (2.5 [0.3] vs 3.3 [0.6]; P<.0001) and location in distal segments (44.5% vs 29.4%; P<.0001). Primary deployment (85.5% vs 95.5%; P<.0001) and postdilatation success rates (1.9% vs 4.8%; P=.039) were higher in group 2, with no differences in vessel dissection rate (4.7% vs 4.4%; P=.85). Direct stenting with 2.5 mm stents was associated with a lower success rate than larger stents. Vessel tortuosity, angulation, calcification, lesion severity and distal location were also associated with a higher failure rate. The predictive power of our model was 0.87 (95% CI, 0.82-0.92). PMID- 14746723 TI - [Transient left ventricular apical ballooning without coronary stenoses syndrome: importance of the intraventricular pressure gradient]. AB - A syndrome of transient apical ballooning without coronary stenosis, which mimics acute myocardial infarction, was recently described. Although several possible etiologic mechanisms have been proposed and investigated, the precise cause remains unclear. We describe 3 cases of transient left ventricular apical ballooning without coronary stenosis, and discuss the etiology of this syndrome, in particular the possible role of a transient intraventricular gradient. PMID- 14746724 TI - [Effects of cisapride on QT interval in children]. AB - This prospective study evaluated the effects of cisapride on corrected QT interval (QTc) in infants and children. From October 2000 to March 2003 two electrocardiograms (ECG) were obtained for 175 children (ranging in age from 1.5 months to 16.8 years), before and after 15 days of treatment with cisapride (0.2 mg/kg/dose, 3-4 times/day). A single posttreatment ECG was also obtained for 24 patients (ranging in age from 1.5 month to 15.8 years). No statistically significant differences were found between the mean QTc interval before (0.390 [0.018 s]) and after treatment (0.391 [0.018 s]). In patients for whom only a posttreatment ECG recording was performed, mean QTc interval was 0.399 (0.018 s). The QTc interval was never longer than 0.450 s in any of the children. In our experience the use of cisapride at therapeutic doses in infants and children who have no associated risk factors does not significantly prolong QTc interval. PMID- 14746726 TI - [Management of end-stage lung disease]. PMID- 14746727 TI - [Exercise limitation in patients with chronic obstructive pulmonary disease at the altitude of Bogota (2640 m). Breathing pattern and arterial gases at rest and peak exercise]. AB - OBJECTIVE: To describe the response to exercise of normal subjects and patients with chronic obstructive pulmonary disease (COPD) in Bogota, Colombia (altitude: 2640 m; atmospheric pressure: 560 mm Hg) and compare it with data published on COPD patients at sea level. Healthy people increase their minute ventilation to attenuate hypoxemia (PaCO2: 30 mm Hg; PaO2: 63 mm Hg). MATERIAL AND METHOD: A descriptive study was carried out on healthy subjects and COPD patients. Exercise limitation was determined by an incremental test on a cycle ergometer. RESULTS: The study enrolled 16 healthy subjects and 25 COPD patients (forced expiratory volume in 1 second: 43.3% [SD 13%]). Minute ventilation at rest was greater in COPD patients compared with healthy subjects, it was not adequately sustained during exercise, and there was a reduction in peak oxygen uptake (53.0% [15%]). At peak exercise, inspiratory capacity decreased (-0.62 [0.34] L), the ratio of minute ventilation to maximal voluntary ventilation increased, and severe hypoxemia occurred (PaO2: 49.9 [9.9] mm Hg). There was significant correlation between hypoxemia and the percentage of predicted peak oxygen uptake (r=0.60), leg fatigue (r=-0.62), percentage of predicted peak inspiratory capacity (r=0.61), and the percentage of predicted peak tidal volume (r=0.49). Minute ventilation at rest was shown to be higher, there was a greater reduction in the inspiratory capacity during exercise, and hypoxemia was more severe at rest and during exercise for patients with COPD in Bogota, compared with those at sea level. CONCLUSIONS: Patients with COPD living in Bogota were shown to have lower tolerance to exercise evidenced by ventilatory limitation and severe hypoxemia. Increased minute ventilation at rest, greater reduction in inspiratory capacity, and severity of hypoxemia during exercise were the main differences between COPD in Bogota and at sea level. PMID- 14746728 TI - [Lung function in cardiac dysfunction]. AB - OBJECTIVE: The alterations in lung function caused by heart failure were first described some decades ago. The advent of new tools for the diagnosis and investigation of heart disease, such as echocardiography, has subsequently made it possible to classify cardiac dysfunction with greater precision. The objective of this study was to analyze and compare a series of lung function and gas exchange variables in patients who had been classified into 4 groups according to type of heart disease as diagnosed by echocardiography. MATERIALS AND METHOD: Emergency room patients whose main symptom was acute dyspnea caused by cardiac or respiratory disease were included in the study. The final sample comprised 71 patients whose echocardiogram revealed cardiac dysfunction. Spirometry was carried out and resting arterial blood gases measured in this group. RESULTS: Of the 71 patients with cardiopathy, 31 had systolic dysfunction, 27 diastolic dysfunction, 7 cor pulmonale, and 6 primary valve disease. Spirometry revealed a generally obstructive pattern, more marked in the group with cor pulmonale. Analysis of arterial blood gases revealed slight hypoxemia with normocapnia in all groups, but this was more accentuated in the patients with cor pulmonale and diastolic dysfunction. An analysis of the correlations (Pearson's r) between cardiac and pulmonary variables revealed the statistically significant associations between cardiac mass and other variables to be as follows: forced vital capacity r=0.34 (P=.02), forced expiratory volume in one second r=0.526 (P=.0001), forced expiratory volume in one second as a percentage of predicted r=0.3 (P=.037), and forced midexpiratory flow rate r=0.31 (P=.03). The correlation between left ventricular ejection fraction and PaO2 was r=-0.312 (P=.01); the correlation between left ventricular end-diastolic diameter and PaO2 was r=0.369 (p=.006). CONCLUSIONS: In patients with cardiac dysfunction, spirometry reveals a generally obstructive pattern, which is more accentuated in patients with right ventricular dysfunction owing to the existence of prior lung disease. The associations found between the cardiac and lung function variables do not help the physician to determine the predominant diagnosis for a patient more precisely or to establish a prognosis. PMID- 14746729 TI - [Thoracoscopic sympathectomy for palmar hyperhidrosis. Immediate results and postoperative quality of life]. AB - OBJECTIVE: Primary palmar hyperhidrosis is a socially and occupationally debilitating disorder characterized by excessive sweating. The purpose of this study was to evaluate the results, complications, and degree of satisfaction among patients who underwent video-assisted bilateral thoracoscopic sympathectomy of the second and third ganglia (T2-T3) to treat primary palmar hyperhidrosis at the Department of General and Thoracic Surgery of the Hospital Universitario Virgen Macarena in Seville, Spain. MATERIAL AND METHOD: A study of 226 thoracoscopic sympathectomies was undertaken based on case histories and a prospective pre- and postoperative questionnaire survey. The sample was composed of 113 patients (47 males and 66 females), ranging in age from 14 to 50 years, with primary palmar and axillary hyperhidrosis, in some cases severe. Bilateral video-assisted thoracoscopic T2-T3 sympathectomy was performed under general anesthesia in all cases. Follow up included a questionnaire on pre- and postoperative quality of life and degree of satisfaction. Descriptive statistics on the surgical procedure, quality of life, and postoperative changes were compiled and frequency analyzed. A nonparametric Wilcoxon test for paired variables was performed to contrast significant differences between pre- and postoperative quality of life related to hyperhidrosis and its complications. RESULTS: The therapeutic success rate was 100%. Complications were seen in 14.2% of the cases and included hemothorax in two, hemopneumothorax in three, pleural hemorrhage in two, and minimal apical airspace in nine. Of the 106 patients who were monitored over a period of 6 to 12 months through follow-up interviews and questionnaires, 67% developed compensatory sweating, 95% reported improvement in quality of life, and 4% experienced no change in quality of life, mainly because of the emergence of compensatory sweating. Of the patients interviewed, 97.2% said that they would undergo the operation again. CONCLUSION: Video-assisted thoracoscopic sympathectomy for the treatment of primary palmar hyperhidrosis is effective, with low rates of morbidity and no mortality. Despite the appearance of postoperative changes such as compensatory sweating, patient satisfaction with the procedure is high and their quality of life improves. PMID- 14746730 TI - [Costs of chronic obstructive pulmonary disease in Spain. Estimation from a population-based study]. AB - OBJECTIVE: In the studies carried out to date, the cost of chronic obstructive pulmonary disease (COPD) may have been overestimated due to the inclusion of previously diagnosed patients seeking medical attention for their symptoms. As a result, the severity of the cases included in these studies may have been greater than in an unselected sample of the general population. The aim of the present study was to estimate the direct cost of COPD on the basis of a representative sample of the overall Spanish population between 40 and 69 years of age (from the IBERPOC study). METHOD: The cost was evaluated retrospectively by means of a questionnaire completed by the 363 patients with COPD from the IBERPOC study with questions referring to the previous year. Standardized spirometry was performed on all the patients by a pneumologist in each of the 7 geographical areas in which the study was carried out. RESULTS: Hospitalization accounted for the greatest expenditure (41% of total), followed by drug therapy (37%). The cost was euro;98.39 per patient, and euro;909.50 per previously diagnosed patient. The cost per person of severe COPD was more than 3 times that of moderate COPD and more than 7 times that of mild COPD. The estimated annual cost of COPD in Spain was euro;238.82 million (for 1997). CONCLUSION: The present study, which was the first to estimate the cost of COPD in a representative sample of the general population, found the cost to be lower than in studies analyzing samples of patients with previous diagnoses of COPD. The cost distribution is not in line with recommended health care practices, underlining the need to optimize resources used to monitor and treat the disease, with an emphasis on early diagnosis. PMID- 14746731 TI - [Evaluation exercise tolerance in COPD patients: the 6-minute walking test]. PMID- 14746732 TI - [Lung preservation: current practices]. PMID- 14746733 TI - [Primary pulmonary lymphoma presenting as a pulmonary mass with cavitation]. AB - Primary pulmonary lymphoma is a rare entity usually formed of B-type cells, usually low-grade and composed of mucosal- or bronchial-associated lymphoid tissue. High-grade primary pulmonary lymphomas usually occur in immunodeficient patients who mostly present with respiratory and nonspecific symptoms. A chest x ray may show a pulmonary mass or atelectasis and pleural effusion. In such cases, the prognosis is worse than for low-grade pulmonary lymphomas; survival is 8 to 10 years and there is a higher probability of local progression or metastasis. We report the case of an immunocompetent 76-year-old patient who had a pulmonary mass with cavitation secondary to a large B-cell primary pulmonary lymphoma. After the fourth session of chemotherapy the pulmonary mass was reduced in size and an aspergilloma was seen to have developed in the residual cavity. A review of the literature revealed this case to be anecdotal as it is extremely infrequent for a primary pulmonary lymphoma to present in the form of a single mass with cavitation and with few symptoms. PMID- 14746734 TI - [Surgical treatment for compensatory hyperhidrosis in Adie syndrome]. AB - Ross syndrome is characterized by a triad of tonic pupil, areflexia and segmental hypohidrosis. Hypohidrosis may be accompanied by contralateral hyperhidrosis, probably due to a compensatory mechanism. We report a case of Ross syndrome with socially disabling left-sided hyperhidrosis. Sympathectomy of the second and third thoracic ganglia was performed with satisfactory results. With excellent results in primary hyperhidrosis and very low morbidity, thoracic sympathectomy is the definitive treatment for hyperhidrosis. PMID- 14746735 TI - [Solitary plasmacytoma in a rib]. PMID- 14746736 TI - [Hypoxemia from multiple causes in a patient with acquired immunodeficiency syndrome]. PMID- 14746737 TI - [HIV/AIDS infection and primary care]. PMID- 14746738 TI - [Written information on the use of aerosols in COPD patients. Can we improve their use?]. AB - OBJECTIVES: To determine whether the use of written information alone improved inhalation technique with pressurized canister inhalers in patients with chronic obstructive pulmonary disease (COPD). To compare the effectiveness of written information versus verbal explanation provided by nursing personnel on the use of inhalers. DESIGN: Interventional study. SETTING: Urban health center. PARTICIPANTS: 120 patients with COPD treated with pressurized canister inhalers. INTERVENTIONS: The participants were divided randomly into three groups of 40 patients each. In one group no intervention was used, in the second group verbal explanations were provided, and in the third group written information was provided. MAIN MEASURES: We recorded percentage compliance with 5 criteria for the correct use of inhalers at the start of the study and 3 months after the intervention in all groups. RESULTS: Initially, performance of the inhalation technique by patients with COPD was poor (mean compliance 40%). Performance improved significantly in both intervention groups, with no significant difference between them. Final mean compliance was 74% in the written information group and 82% in the verbal information group. CONCLUSIONS: The use of written information about the use of inhalers for patients with COPD significantly improved utilization to a degree similar to that obtained with verbal explanations. PMID- 14746739 TI - [Interventional studies of inhalation technique in patients with chronic obstructive pulmonary disease have methodological drawbacks, but are nonetheless necessary]. PMID- 14746740 TI - [Mortality due to asthma in the city of Barcelona (1983-1993)]. AB - OBJECTIVE: To find asthma mortality in the city of Barcelona. DESIGN: Descriptive study of mortality. SETTING: City of Barcelona. MAIN MEASUREMENTS: Deaths due to asthma in the city of Barcelona in the period 1983-1993 were studied through the register of mortality at Barcelona's Municipal Institute of Health, which in turn is supplied by the Statistical Gazette of Deaths. Rates of mortality per 100,000 inhabitants were calculated, overall and broken down by sex and by age. The ratio of mortality comparing city districts and the place and season of decease was also worked out. RESULTS: There were 716 deaths due to asthma (overall rate of 3.82/100,000 inhabitants; 3.3 in men and 4.33 in women). Almost two-thirds of deaths occurred in people over 65. Mortality was stable in the entire period except in the over-65s, in which a downwards trend was discerned (beta=-0.63; P=.037). For the 5-34 year old group, the rate oscillated between 0.1 and 0.6/100,000 inhabitants. The number of deaths in the over-65s was greater in winter (31.7%; 95% CI, 27.8-35.7). 56.2% of deaths occurred at home. Hospital deaths were more common among women (P<.001) and the under-65s, and their trend is upwards (P=.004). CONCLUSIONS: Asthma mortality in the city of Barcelona was stable during the period studied. Its rate for the 5-34 year-old age group was higher than for Spain and slightly greater than in similar nearby countries. PMID- 14746741 TI - [Lifestyle and metabolic control in patients with type 2 diabetes mellitus. Construct validation of IMEVID questionnaire]. AB - OBJECTIVE: To evaluate construct validity of an instrument to measure lifestyle in patients with type 2 diabetes mellitus (IMEVID questionnaire) DESIGN: Comparative cross-sectional study. SETTING: Six family medicine units (primary care). PATIENTS: 412 adults with type 2 diabetes. MEASUREMENTS: The IMEVID was applied by self-administration and the following metabolic control parameters values were measured: body mass index (BMI), waist/hip index (WHI), hemoglobin A1C (HbA1C), total cholesterol (TC), triglycerides (TG), systolic blood pressure (SBP), diastolic blood pressure (DBP), and the average of fasting plasma glucose in the last three months (FG). The test hypothetic construct was that higher total scores in the IMEVID (better lifestyle) would be associated with lower values in the metabolic control parameters. Three groups of subjects were formed based on the total score: group one (quartile 75). Differences on these parameters between groups were researched. RESULTS: 389 subjects completed the study, 69,2% were women. Seven IMEVID domains had weak correlations at least with two of eight parameters (r between 0.22 and 0.16; P<.05). The total score had correlation with six of eight parameter (r between 0.18 and 0.10; P< or =.05). The group three subjects had lower levels of BMI, WHI, HbA1C, FG, TC and TGL than group one subjects (P<.05). CONCLUSIONS: IMEVID has construct validity to measure the lifestyle in subjects with DM2. Its total score discriminates outstanding clinical characteristics in these patients. PMID- 14746742 TI - [Repercussions on primary care of Act 41/2002 on patient autonomy, rights to information and clinical documents (II)]. PMID- 14746743 TI - [Characteristics of the demand for medical attention in primary care of a health area in the Community of Valencia (SyN-PC study)]. AB - OBJECTIVES: To quantify with primary data the various kinds of consultations, the reasons for them and the diagnoses; to measure preventive actions and diagnoses using machines. To check the recording of the kinds of consultations and reasons for them in the clinical notes and on the doctor's work-sheet. DESIGN: Observation, cross-sectional and multi-centre study. Clinical care activity was measured on two January days in 2002. SETTING: All the health centres in Area 17 of the Community of Valencia. PARTICIPANTS: Representative sample of 2051 patients belonging to 20 primary care medical clinics from the above area, selected by sampling stratified by health centres. MAIN MEASUREMENTS: The activity of a working day was recorded and structured as follows: a) care activity (scheduling, age, sex, records code, kind of consultation and motive, specifying whether this was care activity, preventive or bureaucratic); b) activity caused by main reason for consultation (anamnesis, examination and further tests); c) plan of action (hygienic and dietary measures, medication and referrals); d) use of the clinical records according to SOAP sections; e) interruptions during the consultation. RESULTS: Study of care activity and its correlation with the actions recorded in the clinical record with primary data enables the reality of health care to be fairly faithfully perceived. These results highlighted the usefulness of clinical record audits to measure health care delivery and to identify patterns of consumption of health resources, as a necessary pre-condition of more efficient primary care management. PMID- 14746744 TI - [Vaccinations and pregnancy (I): vaccines that are indicated for pregnant women]. PMID- 14746746 TI - [Survey on the prescription of generic medicines in primary care]. PMID- 14746747 TI - [Secondary prevention of ischaemic cardiopathy: are we doing it right?]. PMID- 14746748 TI - [Syndrome of confusion as side-effect of cycloplegic collyrium]. PMID- 14746749 TI - [Congenital myotonic dystrophy in a rural health area]. PMID- 14746750 TI - [High temperature and ankylosing spondylitis]. PMID- 14746751 TI - [Type-2 diabetes mellitus: does insulin provision in primary care improve metabolic control?]. PMID- 14746752 TI - [Diabetes type 2: the misunderstood disease]. PMID- 14746753 TI - [Factors related to the tuberculous patient and the study of contacts]. AB - CONTEXT: While the maximum priority of the programs for tuberculosis prevention and control is the identification and treatment of all the people with active tuberculosis, the second priority is the evaluation of their contacts. The goal of this study has been to evaluate the factors of the tuberculous patients associated with the study of their contacts. PATIENTS AND METHODS: The incident cases of tuberculosis occurred during the period 1992-1997 in the province of Lleida were studied. The variables of the study were: age, sex, place of residence, infection by HIV, parenteral drug use (PDU), excessive alcohol consumption, anatomical location of tuberculosis, existence of cavernous lesions in the chest X-ray, and microbiological result of the samples of respiratory secretions. The association of the dependent variable (study of the contacts in the incident cases of tuberculosis) with the rest of independent variables it was determined by the raw and adjusted odds ratio. RESULTS: The prevalence of tuberculosis patients whose contacts were studied was 60.4% (63.1% in the bacilliferous cases and 51.9% in the non-bacilliferous cases, p=0.003). A higher number of patients whose contacts were studied was detected between those that were not HIV-infected (ORa: 4.53; 95% CI: 2.91-7.05), between those with lung as the anatomical localization of the tuberculous disease (ORa: 2.62; 95% CI: 1.70 4.05) and between those that did not showed excessive alcohol consumption (ORa: 1.69; 95 CI: 1.17-2.46). CONCLUSIONS: The global prevalence of tuberculous whose contacts are studied should be increased, especially among the patients with sociosanitary risk (HIV, PDU and alcoholism), and among the patients with higher probability of transmission. PMID- 14746754 TI - [Syncope: many studies and few diagnoses?]. AB - INTRODUCTION: Syncope is a frequent disorder shows diagnostic difficulties, so that an important percentage of patients remain without diagnosis after the implementation of diagnostic tests that sometimes are unnecessary. METHOD: A retrospective analysis of the patients admitted during a period of two years because of syncope in an Internal Medicine Service was carried out. RESULTS: Of a total of 2,878 patients hospitalized during the years 1999 and 2000, 79 were admitted because of syncope (2.74%). Forty-five (57%) were men and 34 (43%) were women, with a median age of 68.8 years. Twenty-seven patients (34%) had history of syncope. After a median hospitalization of 9.5 days, the etiological diagnosis was established in 53 (67%) patients: in 25 patients (31.6%) the syncope was mediated by neurological mechanisms, in 16 (20.25%) it was of cardiology origin, in 4 (5.06%) it was of neurological origin and in 8 (10.1%) the syncope was secondary to other causes. In 54 patient (68.8%) at least one cardiology test was carried out (41 Holter, 32 echocardiograms, 6 tilt board tests, 3 maneuvers of massage of the carotid sinus, 3 ergometry studies, 3 electrophysiological studies, 2 coronary angiographies and 3 ventilation perfusion lung gammagraphies). In 51 patient (64%) at least one neurological test was carried out (31 EEG, 42 CT, 2 MNR, 1 Doppler cranial ultrasound and 3 electroneuromyogram). Only 4 of the Holter carried out were diagnostic, as well as 3 of the echocardiograms and only 2 of the CT carried out. No EEG was useful for the etiological diagnosis of the syncope. CONCLUSIONS: There is an excessive utilization of some diagnostic tests (EEG, Holter, cerebral CT), in spite of which in an important percentage of patients with syncope the etiological diagnosis is no done (33%). The use of diagnostic algorithms based on detailed clinical history, physical exploration and assessment of the ECG is proposed, that they would make it possible to issue a diagnostic hypothesis from which there could be begun the diagnostic study with a use as rational as possible of the complementary examinations. PMID- 14746755 TI - [Intestinal parasitosis in the asymptomatic Subsaharian immigrant population. Gran Canaria 2000]. AB - CONTEXT: A prospective study has been conducted in order to known the prevalence and the type of intestinal parasitoses in a cohort of healthy Subsaharian immigrants that reached Gran Canaria along the year 2000. METHODS: 348 fecal samples of 121 immigrants have been studied. In each patient a minimum of two samples and a maximum of three samples was studied. Each sample was evaluated through two techniques: visualization of a direct preparation dyed with Lugol and through the technique of concentration of Kato. RESULTS: 23.1% of the patients presented pathogenic parasites in feces, being observed 2 parasites in 5 of the patients (17.8%). The geohelminths (Ancylostoma duodenale/Necator americanus, Ascaris lumbricoides, Trichuris trichiura and Strongyloides stercolaris) were the parasites more frequently found (87.9%). Specifically, the parasitosis found in the greatest number of participants was hookworm disease (44.8%). In this study the effectiveness of the evaluation of three feces samples is demonstrated in order to increase the percentage of diagnosed intestinal parasitoses. The visualization of a direct preparation dyed with Lugol permitted the diagnosis of 63.6% of the parasitoses, while the technique of Kato detected 56.6% of the helminth infections. CONCLUSIONS: The prevalence of geohelminths in the asymptomatic Subsaharian immigrant population is elevated. The complementary use of the microscopic study of a direct preparation dyed with Lugol and the technique of Kato increase the diagnostic performance. PMID- 14746756 TI - [Pathological characteristics of patients with diabetes mellitus type 2, in Spanish Primary Care]. AB - CONTEXT: To know the characteristics, related risk factors, and degree of control in patients with diabetes mellitus type 2 (DM2) in our country. METHODS: Observational, unrandom, opened, and multicenter study. Anthropometric characteristics, substance abuse, medication, control of diabetes, cholesterol, and triglycerides were analyzed in 5,395 patients. The patients were classified according to the length of the diagnosis: recent diabetes (RD) and known diabetes (KD). The chi-square test was utilized in order to compare the categorical variables, and the Student's t test for compare the continuous variables. The relationship between these variables was analyzed through the Pearson's correlation coefficient, and an analysis of multiple correspondence was carried out. RESULTS: Median age, 63 years; obese, 34%; smokers, 11%; drinkers, 24%; hypertensives, 44%; lipemic, 42%. Control degree: HbA1c>6.5% in 79%, LDLc>115 mg/dl in 80%. Hypoglycemic treatment: sulfonylureas, 62.8%; antidiabetics combination, 5%; only insulin, 4.3%; insulin combined with antidiabetics, 20.6%. Control of diabetics, of lipids, and of weight was lower in the patients with KD that in the patients with RD (HbA1c, 7.6% versus 7%; LDLc 148 mg/dl versus 136 mg/dl; percentage of obese, 27.2% versus 38.62%). A relationship between the length of evolution of diabetes and the deterioration of the control of blood glucose and of lipids was detected. CONCLUSION: DM2 and its associated risk factors are insufficiently controlled in our country. The drug combination and insulin are utilized scarcely. PMID- 14746757 TI - [A 48 year old man with recurrent abdominal pain for 8 months, anemia and weight loss]. PMID- 14746758 TI - [Giant hepatomegaly with calcifications]. PMID- 14746759 TI - [Hypopigmented cutaneous lesions in a black immigrant]. PMID- 14746761 TI - [Tendinitis by quinolones: treatment and drug class effect in two new cases]. PMID- 14746760 TI - [Usefulness of miglitol in patients with diabetes mellitus type 2 and insufficient control of the blood glucose]. AB - OBJECTIVE: Miglitol belongs to the group of the inhibitors of the alpha glucosidases and in several studies has demonstrated its usefulness in the improvement of the glycemic control in the different types of diabetic patients. The goal of our study was to analyze the usefulness of miglitol in the blood glucose and lipid control in a group of patients with diabetes type 2 treated with sulfonylureas and insulin, with insufficient glycemic control. PATIENTS AND METHODS: A group of 33 patients with diabetes mellitus type 2 treated with sulfonylureas and insulin and with insufficient glycemic control (HbA1c>7.5%) was studied. All patients were treated with miglitol during 3 months with progressive doses: the first week 50 mg twice a day, subsequently and during one month 50 mg three times a day. At the beginning of the study and up to three months the following variables were measured; weight, height, BMI (body mass index), systolic and diastolic blood pressure, HbA1c, number of episodes of peripheral hypoglycemia, basal glucose, albuminuria, total cholesterol, LDL-cholesterol, HDL cholesterol and triglycerides, as well as transaminases (GOT, GPT, gamma GT and bilirubin). RESULTS: The median age was 64.1 +/- 12.5 years, and the median length of diabetes mellitus was 9.1 +/- 7 years. The median value of BMI was 27.7 +/- 6.9 kg/cm2. The blood glucose and HbA1c values decreased 4.8% and 5.8%, respectively. A decrease in the number of hypoglycemia episodes (39.4% previous quarter versus 3% quarter wih miglitol) was observed. Also decreased the dose of sulfonylureas needed by the patients (86.2 +/- 24.3 mg/day versus 64.6 +/- 21.9 mg/day; p<0.05) (25%). Total cholesterol, HDL-cholesterol, and LDL-cholesterol levels were not modified, but indeed a reduction of the levels of triglycerides (145.2 +/- 111 mg/dl versus 133.1 +/- 79 mg/dl; p<0.05) (8.3%) was detected. Fifteen percent of patients showed side effects (digestive discomfort) that disappeared two or three weeks after beginning the treatment. CONCLUSIONS: Miglitol is a useful drug in order to improve the glycemic control in diabetic type 2 patients treated previously with other drugs and with insufficient control of the blood glucose. Its side effects are limited and transitory. PMID- 14746762 TI - [Non-pharmacological methods in tobacco dishabituation]. PMID- 14746763 TI - [Troponins in its context: from the test tube to the clinical arena]. PMID- 14746764 TI - [Indigenous amebic liver abscess. Presentation of a case]. PMID- 14746765 TI - [Extrapulmonary sarcoidosis as the etiology in a patient with fever of unknown origin]. PMID- 14746766 TI - [Primary epiploic appendix inflammation]. PMID- 14746767 TI - [Hypomagnesemia and cardiac failure]. PMID- 14746768 TI - The effect of milk production level on host resistance of dairy cows, as assessed by the severity of experimental Escherichia coli mastitis. AB - This study investigated the possible effects of milk production level on the host resistance of dairy cows. High (n = 18) and low (n = 18) producing cows on a research farm, which respectively produced 11 443 and 7 727 kg milk in their previous lactation, were compared. To enhance the possible differences in host resistance between high and low producing cows, the animals in both groups were metabolically stressed by overfeeding during the dry period or were fed according to requirements, resulting in four groups of nine cows. The metabolic status was monitored from two weeks pre-partum until 2.5-4.5 weeks post-partum. Host resistance was assessed by measuring the severity of experimentally induced Escherichia coli mastitis. Pre-partum blood glucose levels tended to be higher in overfed cows than in cows fed according to requirements. The post-partum energy balance was significantly more negative in high producing cows than in low producers, and tended to be more negative in overfed cows compared to cows fed according to the requirements. Post-partum plasma glucose, NEFA, beta-OH-butyrate and urea concentrations were similar in the four groups. Plasma glucose concentrations were significantly lower and liver triacylglycerol concentrations were significantly higher in third than in second parity cows. Host resistance was not affected by the production level or feeding regimen. There were no significant correlations between the metabolic status and the severity of experimental E. coli mastitis, except for the relatively more severe mastitis in the cows with beta-OH-butyrate concentrations above 1.4 mmol/L. In conclusion, milk production level did not affect host resistance in dairy cows, as measured by the severity of experimental E. coli mastitis. Even in a situation where cows were metabolically stressed by overfeeding, high producers were as able as low producers to cope with the demands of milk production, without consequences for host resistance. PMID- 14746769 TI - Listeria monocytogenes contamination of finishing pigs: an exploratory epidemiological survey in France. AB - Listeria monocytogenes is a foodborne pathogen of major concern for public health in industrialised countries. Since L. monocytogenes carriage by pigs at the herd level could be a primary source for carcass contamination, control measures should be designed to reduce the L. monocytogenes load at the pre-harvest stage. For this purpose, an exploratory analytical survey was carried out in 2000-2001 in 93 French farrow-to-finish pig farms concerning L. monocytogenes contamination in pigs before they left for the slaughterhouse. On each farm, the L. monocytogenes status of a batch of contemporary fattening pigs housed in the same room was assessed on faecal material samples taken by means of gauze swabs wiped on the perianal region of the pigs. Fourteen percent of the batches studied had at least one contaminated sample and were therefore classified as L. monocytogenes contaminated batches. Two logistic regression models were used to assess the association between managerial and hygiene practices and the risk of L. monocytogenes contamination of the batch at the end of the finishing period on the whole data set (n = 93) and in the wet feeding farms only (n = 57). Wet feeding during the fattening period was identified as a risk factor for L. monocytogenes contamination. Risk factors related to the introduction of L. monocytogenes in pig facilities were identified for both the general and wet feeding farm data sets. Poor care paid to hygiene on the farms was found to increase the risk of being infected (boots cleaning, change room presence). When the duration of the empty period prior to the introduction of growing pigs was less than one day in the fattening section, the risk of L. monocytogenes contamination was significantly increased. For wet feeding farms, a distribution pipeline cleaning procedure including disinfection was found to be associated with a higher risk of contamination than no cleaning or a procedure consisting of rinsing with water only. PMID- 14746770 TI - Descriptive spatial analysis of BSE in western France. AB - The spatial heterogeneity of Bovine Spongiform Encephalopathy (BSE) was analysed on the 84 cases confirmed in western France (WF) between August and December 2000, when both the Mandatory Reporting System and an active surveillance on cattle at risk were running. Ninety-four percent of these cases were born between June 1993 and June 1996, and we analysed the location at birth. One disease mapping and two clustering methods (Scan of Kulldorff and the method of Besag and Newell) were used. In order to attenuate the contrasts artificially created by the standard disease mapping method (over-dispersion), we estimated the Standard Incidence Ratio (SIR) with a Bayesian method (Poisson-Gamma model) allowing a smoothing of the estimators. The geographical location of interest was the "canton", that divided the total area into 526 geographical units. The background population (2.6 million cattle) was obtained from the Agricultural Census 2000. We tested the hypothesis of a homogenous spatial distribution of the BSE risk where the expected number of BSE cases per unit area was obtained by applying the overall BSE rate in WF to each "canton", standardised on the type of breed, dairy versus beef suckler. The SIR ranged from 0.80 to 2.18 and the spatial distribution of BSE cases was significantly heterogeneous. Two spatial clusters were detected with the spatial scan statistics of Kulldorff and the method of Besag and Newell (18 to 20 observed BSE-cases per cluster with a radius of 45 km) centred on the "departement" of Cotes-d'Armor and Mayenne. Another cluster was detected with the method of Besag and Newell (9 observed BSE-cases) in the "departement" of Finistere. The results proved that the risk of BSE is linked to the geographical location in the area of the study. PMID- 14746771 TI - Antigenic and genetic characterisation of lipoprotein lppC from Mycoplasma mycoides subsp. mycoides SC. AB - Lipoprotein lppC, an immunodominant antigen, and its corresponding gene lppC were characterised in Mycoplasma mycoides subspecies mycoides small colony (SC) type, the etiological agent of contagious bovine pleuropneumonia (CBPP). The lppC gene was found in the type strain of M. mycoides subsp. mycoides SC and in field strains isolated in Europe, Africa, and Australia, as well as in vaccine strains. Southern blot analysis indicated the presence of at least four copies of lppC in the genome of M. mycoides subsp. mycoides SC, of which only one seems to be functional. Genes homologous to lppC have also been detected in closely related mycoplasmas such as M. mycoides subsp. mycoides large colony (LC) type and in M. sp. bovine group 7. lppC is encoded as a precursor with a consensus sequence for a prokaryotic signal peptidase II. The amino acid sequence of lppC and its precursor showed similarity to both LppB (at the N-terminal domain) and LppQ (at the C-terminal domain), two lipoproteins described previously in M. mycoides subsp. mycoides SC. The N-terminal domain of the mature lppC seems to be surface exposed. The C-terminal domain presented an integral membrane structure made up of five repeated units, rich in hydrophobic and aromatic amino acids, which may have pore forming potential in the mycoplasmal membrane. A recombinant peptide representing the N-terminal half of lppC was obtained following cloning in vector pETHIS-1 and expression in Escherichia coli hosts. The recombinant protein was used on immunoblots for serological analysis of sera from cattle that were naturally or experimentally infected with M. mycoides subsp. mycoides SC. PMID- 14746772 TI - Comparison of genome segments 2, 7 and 10 of bluetongue viruses serotype 2 for differentiation between field isolates and the vaccine strain. AB - Bluetongue (BT) virus serotype 2 (BTV 2) was first confirmed in Tunisia in February 2000 and has since spread northward and westward, infecting several other countries and islands, including Corsica, where clinical disease was reported in October 2000. BT was again reported on the Island in July 2001, some six months after a vaccination campaign against BTV 2. The molecular relationship between isolates of the BTV 2 Corsican wild-type viruses from 2000 and 2001, and the attenuated BTV 2 vaccine were determined by comparing corresponding sequences of genome segments 2, 7 and 10 with each other and with already published sequences available in the genome database. Complete genetic stability was observed between the isolates of the Corsican BTV 2. There was some divergence between the nucleotide sequences of segment 10 obtained from the wild-type and vaccine virus strains. Based on these differences, primers were selected that could be used in RT-PCR to differentiate between the wild-type and the vaccine viruses. PMID- 14746773 TI - Cross-species reactivity of seven monoclonal antibodies with equine lymphocytes by flow cytometry. AB - The recognition of equine lymphocyte antigens by monoclonal antibodies (mAbs) directed against human CD11a, CD18, CD21, CD23, CD29 and DR, as well as mouse CD23 was studied by flow cytometry. Unlike anti-CD11a, -CD21, -CD23 and DR mAbs, anti-CD18 and CD29 mAbs labelled the same percentage of horse peripheral blood lymphocytes (PBL) as human PBL. Double-staining with anti-horse immunoglobulin antibodies showed that anti-CD21 and -CD23 mAbs are mainly bound to peripheral blood B lymphocytes. The seven mAbs were also tested on the lymph node and thymus cells. The molecular targets of anti-CD11a, CD18 and CD29 mAbs were confirmed by immunoprecipitation of the membrane proteins. Our results suggest that anti-CD18, -CD29 and -DR mAbs recognise similarly expressed molecular homologues on equine cells, but that anti-CD11a, -CD21 and -CD23 mAbs recognise either different molecules or homologues that are expressed at different levels on horse cells. PMID- 14746774 TI - Evaluation of a procedure to assess the adverse effects of illicit drugs. AB - The assessment procedure of new synthetic illicit drugs that are not documented in the UN treaty on psychotropic drugs was evaluated using a modified Electre model. Drugs were evaluated by an expert panel via the open Delphi approach, where the written score was discussed on 16 items, covering medical, health, legal, and criminalistic issues of the drugs. After this face-to-face discussion the drugs were scored again. Taking the assessment of ketamine as an example, it appeared that each expert used its own scale to score, and that policymakers do not score deviant from experts trained in the medical-biological field. Of the five drugs evaluated by the panel, p-methoxy-metamphetamine (PMMA), gamma hydroxybutyric acid (GHB), and 4-methylthio-amphetamine (MTA) were assessed as more adverse than ketamine and psilocine and psilocybine-containing mushrooms. Whereas some experts slightly adjusted during the assessment procedure their opinion on ketamine and PMMA, the opinion on mushrooms was not affected by the discussion held between the two scoring rounds. All experts rank the five drugs in a similar way on the adverse effect scale i.e., concordance scale of the Electre model, indicating unanimity in the expert panel with respect to the risk classification of these abused drugs. PMID- 14746775 TI - Oral-to-inhalation route extrapolation in occupational health risk assessment: a critical assessment. AB - Due to a lack of route-specific toxicity data, the health risks resulting from occupational exposure are frequently assessed by route-to-route (RtR) extrapolation based on oral toxicity data. Insight into the conditions for and the uncertainties connected with the application of RtR extrapolation has not been clearly described in a systematic manner. In our opinion, for a reliable occupational health risk assessment, it is necessary to have insight into the accuracy of the routinely applied RtR extrapolation and, if possible, to give a (semi-)quantitative estimate of the possible error introduced. Therefore, experimentally established no-observed-adverse-effect-levels for inhalation studies were compared to no-adverse-effect-levels predicted from oral toxicity studies by RtR extrapolation. From our database analysis it can be concluded that the widely used RtR extrapolation methodology based on correction for differences in (estimates of) absorption is not generally reliable and certainly not valid for substances inducing local effects. More experimental data are required (from unpublished data or new experiments) to get insight into the reliability of RtR extrapolation and the possibility to derive an assessment factor to account for the uncertainties. Moreover, validated screening methods to predict/exclude the occurrence of local effects after repeated exposure are warranted. Especially, in cases where chemical exposure by inhalation or skin contact cannot be excluded route-specific toxicity studies should be considered to prevent from inadequate estimates of human health risks. PMID- 14746776 TI - Safety determination for the use of bovine milk-derived lactoferrin as a component of an antimicrobial beef carcass spray. AB - Bovine milk-derived lactoferrin (BMDL), an iron-binding glycoprotein, is known to be an effective natural antimicrobial. It is used as a spray, applied electrostatically, to raw beef carcasses to detach bacteria adhering to the surface in order to reduce microbial contamination. The use of BMDL as a component (at not more than 2% by weight) of an antimicrobial spray was determined Generally Recognized As Safe (GRAS) (GRN 67) for three proposed uses (i.e., beef carcasses, subprimals, and finished cuts), provided that the ingredient statement of food products that contain milk-derived lactoferrin identifies the source of the protein. The use of BMDL spray on only beef carcasses (not subprimals or finished cuts) at a level not to exceed 0.20 ml of formulation per kg of beef was determined safe without the requirement of labeling of food products so treated. The two key components of the assessment are: (1) a determination that exogenous lactoferrin exposure (resulting from its application to beef carcasses) is in the range of existing background exposures of lactoferrin as a result of lactoferrin found naturally in beef, and (2) a determination that this potentially small incremental increase in lactoferrin is safe (i.e., there is no reasonable expectation that BMDL will become an allergen under the conditions of its intended use). PMID- 14746777 TI - Methodological approaches for studying pharmaceuticals in the environment by comparing predicted and measured concentrations in River Po, Italy. AB - A predictive approach seems useful to study human and veterinary pharmaceuticals in the environment and provide an idea of overall levels of contamination, so as to restrict monitoring to those molecules which are most likely to represent possible environmental contaminants. Predicted environmental concentrations (PECs) can be calculated by a mass balance approach, while a recent proposal from the European Agency for the Evaluation of Medicinal Products (EMEA) suggests an alternative method for calculating PEC for each pharmaceutical and then focusing further work on molecules with high PEC values. We used the results of monitoring campaigns on the River Po, in Northern Italy, to assess the accuracy of predictive models with measured environmental concentrations (MECs). The comparison indicated that in some cases a refined PEC value can provide a good approximation of the MEC. In other cases PECs substantially differed from the MECs, particularly when there were not enough data to estimate the environmental fate of the molecule. Predictive models might therefore be useful for studying pharmaceuticals in the environment, providing enough experimental data is available on the environmental fate of the molecules. PMID- 14746778 TI - Indicators of uncertainty in chemical risk assessments. AB - For most of the chemical substances that are subject to regulatory and industrial decision making, important toxicological data are missing. The available dataset is often difficult to interpret, and the differences between alternative, scientifically reasonable interpretations may have major impact on decision making. Since such uncertainties are an important factor in decision making it is essential that they be reported in a transparent and understandable way. In order to clarify how toxicologists report uncertainties, 30 risk assessments for one and the same substance (trichloroethylene) from the years 1973-2001 were searched for phrases indicating uncertainty. These phrases can be divided between four categories: contentual, epistemic, conditionalising, and inferential uncertainty indicators. A typology of uncertainty indicators, based on these categories, is proposed. It is concluded that the use of uncertainty indicators in these texts is not transparent and that the development of standardised uncertainty indicators should significantly improve communications both within the scientific community and between scientists and policymakers. PMID- 14746779 TI - Reference dose for perchlorate based on thyroid hormone change in pregnant women as the critical effect. AB - The most relevant data for developing a reference dose (RfD) for perchlorate exposures comes from human epidemiology and clinical studies, supplemented with available and extensive information on experimental animals. Specifically, serum T4 decrease is the critical effect of perchlorate, based on a mode-of-action analysis and the evidence provided by the body of rodent studies on perchlorate. However, no T4 decreases have been observed in human populations following perchlorate exposure at non-therapeutic doses. An RfD of 0.002 mg/kg-day can be derived using an epidemiology study. A freestanding NOAEL of 0.006 mg/kg-day for T4 decrease was identified in children from the epidemiology study. The use of this NOAEL has the advantage of a being identified in a sensitive subgroup, neonates and children. Data are sufficient to estimate an overall uncertainty factor of 3-fold with this NOAEL based on expected differences in toxicokinetics and toxicodynamics between children, and pregnant women and their fetuses, the second identified sensitive subgroup for perchlorate, and concerns about the over iodination of this population. This RfD is supported by a human clinical study using inhibition of iodine uptake in adults as a measurable surrogate for the critical effect of T4 decrease in humans. However, although this latter study has a well-established dose-response curve for inhibition of iodine uptake, even perchlorate doses that result in a 70% inhibition of iodine uptake have no apparent effect on human T4 levels. Thus, the use of this study as the primary basis of the RfD is problematic. Nevertheless, a benchmark dose of 0.01 mg/kg-day was identified in this clinical study, which supports a threshold value of 0.006 mg/kg-day identified by its authors and the RfD of 0.002 mg/kg-day estimated in this paper. PMID- 14746780 TI - Gaucher disease in children: radiology of non-central nervous system manifestations. AB - The radiological findings in paediatric Gaucher disease (GD) are reviewed and future challenges for radiology are discussed. This overview is based on a literature review and our experience of children with GD in one of two national institutions for paediatric GD in the UK. GD is known to progress more rapidly in childhood. Current imaging is mainly suitable for ascertaining the complications of GD. The UK recommendations for routine radiological surveillance are discussed. With enzyme replacement therapy (ERT), which dramatically modifies the course of the disorder, the challenge for radiology in the future is likely to be assessing treatment efficacy rather than the detection of disease complications. Disease manifestations are likely to change in those on ERT and the most notable recent alteration in the disease profile in childhood is the virtual disappearance of the acute bone crisis in this population. PMID- 14746781 TI - Malignant and benign compression fractures: differentiation and diagnostic pitfalls on MRI. AB - The distinction between malignant and benign compression fractures is a common problem in clinical practice. Various imaging techniques (plain radiography, computed tomography, bone scintigraphy) have been used to differentiate these conditions but they are often inadequate in distinguishing the nature of compression fracture. This review illustrates the magnetic resonance imaging (MRI) features of malignant and benign compression fractures with emphasis on the usefulness, limitations and pitfalls of MRI. PMID- 14746782 TI - Bone within a bone. AB - The "bone within a bone" appearance is a well-recognized radiological term with a variety of causes. It is important to recognize this appearance and also to be aware of the differential diagnosis. A number of common conditions infrequently cause this appearance. Other causes are rare and some remain primarily of historical interest, as they are no longer encountered in clinical practice. In this review we illustrate some of the conditions that can give the bone within a bone appearance and discuss the physiological and pathological aetiology of each where known. PMID- 14746783 TI - Imaging features of hepatocellular carcinoma. AB - This pictorial review concentrates on the imaging features of hepatocellular carcinoma as revealed by ultrasonography, computed tomography, magnetic resonance imaging and angiography. Understanding of the pathomorphological characteristics of the disease is important to precise image interpretation. PMID- 14746785 TI - Trainee reporting of computed tomography examinations: do they make mistakes and does it matter? AB - AIM: To determine the accuracy of trainees reporting computed tomography (CT) examinations. MATERIAL AND METHODS: Over a 6-month period a single consultant reviewed all the CT examinations reported by registrars in one radiology department. After recording a provisional registrar report each examination was jointly reviewed by the consultant and registrar. The consultant's opinion was regarded as the gold standard. Data collected included: the error rate, whether an error was significant, leading to a change in patient management, and whether the mistake was a false-negative or positive. RESULTS: Three hundred and thirty one patients were included in the study. There was an overall error rate of 21.5%. A significant error leading to a change in management was made in 10% of reports, and a significant error that did not lead to a change in management was made in 9.3%; 2.1% of reports had insignificant errors; and 69% of errors were false-negatives. CONCLUSION: Registrars make a significant number of errors affecting patient management when reporting CT and ideally all examinations should be reviewed by a consultant. PMID- 14746787 TI - Mammographic casting-type calcification associated with small screen-detected invasive breast cancers: is this a reliable prognostic indicator? AB - AIM: The aim of the present study was to establish whether mammographic casting type calcification associated with small screen-detected invasive breast cancers is a reliable prognostic indicator. METHODS AND MATERIALS: We retrospectively identified 50 consecutive women diagnosed with an invasive cancer less than 15 mm who showed associated casting calcification on their screening mammograms. Controls were identified that showed no microcalcification and were matched for tumour size, histological type and lymph node status. A minimum of 5 years follow up was obtained, noting recurrence and outcome. Conditional and unconditional logistic regression, depending on the outcome variable, were used to analyse the data, taking the matched design into account in both cases. Where small numbers prohibited the use of logistic regression, Fisher's exact test was used. RESULTS: Five deaths from breast cancer occurred out of the 50 cases, of which three were lymph node positive, two were lymph node negative and none were grade 3. None of the 78 control cases died from breast cancer. The difference in breast cancer death rates was significant by Fisher's exact test (p=0.02). Risk of recurrence was also significantly increased in the casting cases (OR=3.55, 95% CI 1.02 12.33, p=0.046). CONCLUSION: Although the overall outcome for small screen detected breast cancers is good, our study suggests that casting calcification is a poorer prognostic factor. The advantage of a mammographic feature as an independent prognostic indicator lies in early identification of high-risk patients, allowing optimization of management. PMID- 14746788 TI - Major complications after percutaneous nephrostomy-lessons from a department audit. AB - AIM: To audit the performance of our percutaneous nephrostomy service by comparing the major complication rate with the standards recommended by the Society of Cardiovascular and Interventional Radiology and the American College of Radiology: major haemorrhage in <4%, and septic shock in <4%. Secondary aims were to identify common sources of errors for quality improvement measures. METHODS: Major complications sustained between January 1997-December 2002 were identified. All cases had been carried out by the interventional radiology service of a large teaching hospital with the assistance of a nurse and radiographer, under fluoroscopic and ultrasound guidance using a Seldinger technique. Existing department protocols specified pre-procedure antibiotics for suspected infected cases and normal coagulation studies. From records and review of case notes pertinent clinical/procedural details and eventual outcome were assessed. The following were particularly noted: adherence to protocols, clinical status at time of procedure, delay in referral, complication sustained, signs of infection or coagulopathy, timing of procedure (in versus out of hours), level of operator and technical faults. RESULTS: Ten of 318 (3.1%) cases sustained a major complication: five had sepsis alone, two haemorrhage (one with sepsis as well) and three patients had a major pelvic injury (one with sepsis as well). Thus the major sepsis and haemorrhage rates were 2.2 and 0.6%, and were within the recommended threshold limits but proportionately more complications occurred out of hours: six of 105 (5.7%) versus four of 312 (1.8%; p=0.087). Sepsis was the most serious complication and may have contributed to the death of two patients. On individual case analysis, failed instrumentation with delay to definitive renal drainage was a common factor with sepsis; but the following were contributory factors in one or more cases: omitted antibiotics (in three of 10; two became septic), technical factors in four cases [medial renal puncture (n=1), damage due to fascial dilator (n=1) or peelaway sheath (n=2)] and delay in diagnosis/therapy (of 1-8 days, in six of 10 cases of whom four out of six became septic). One pelvic injury required surgical correction (contributory factor faulty use of peelaway sheath). Patients with haemorrhage settled with prolonged tube drainage alone. CONCLUSION: An adequately staffed percutaneous nephrostomy service can perform within published clinical standards. Best practice factors identified were: attention to agreed protocols and algorithms, pre-procedure antibiotics, careful renal puncture and care with use of dilators/peelaway sheaths, but the paramount finding was that sepsis was the most serious complication, contributing to death (two of 10 in this study) or a significant increase in the level of care required. The risk is greatest after failed instrumentation (retrograde ureteral stent or percutaneous nephrostomy insertion) and particularly if there is a further delay before establishment of renal drainage. A close working relationship between interventional radiologists and urologists is crucial. PMID- 14746789 TI - Conscious sedation for endoscopic and non-endoscopic interventional gastrointestinal procedures: meeting patients' expectations, missing the standard. AB - AIM: To assess the level of sedation, patient satisfaction and frequency of unplanned events with conscious sedation for interventional procedures. MATERIALS AND METHODS: One hundred and seventeen patients were assessed prospectively before, during and after procedures. Blood pressure, pulse, oxygen saturation and sedation level were monitored and patients followed up after 24 h. Sedation was scored after drugs were given in accordance with an established protocol. Doses were recorded, as were patients' weight, age and ASA grade and any unplanned events and their management. RESULTS: Seventy-six of the 117 patients (65%) had no unplanned event, 20 (17.1%) became agitated, 15 (12.8%) hypotensive, three (2.6%) hypoxic and three (2.6%) had more than one response. Twelve patients required active management. Fifty-two (44.4%) had a sedation level of CAD>CPA, which was consistent with A(2) receptor identification. CAD and NECA induced both endothelium-dependent and -independent relaxations of the aortas. The endothelium-dependent responses to both agents and the independent responses to CAD were significantly attenuated by beta-alanine treatment. The relaxation responses of the aortas from control and taurine deficient rats to CAD and NECA were markedly antagonized by ZM241385 (10(-5) M), suggesting the involvement of A(2A) adenosine receptors. Further, N-nitro-L arginine methyl ester (L-NAME; 10(-5) M) significantly attenuated the endothelium mediated relaxation produced by CAD and NECA in both groups. However, the inhibitory effect of L-NAME was less on the beta-alanine-treated tissues, providing evidence that the effect of taurine deficiency was linked to a reduction in nitric oxide generation. As in the aorta, CAD produced both endothelium-dependent and -independent relaxation responses in the rat superior mesenteric artery, and both responses were inhibited by chronic beta-alanine treatment, suggesting that not only similar responses can be generated by a given adenosine agonist in different vascular beds, but also beta-alanine treatment modulates these responses. On the other hand, while CPA elicited only endothelium independent aortic relaxation, this response was not altered by taurine deficiency. The results indicate that endogenous taurine deficiency causes differential inhibitory effects on adenosine receptor-mediated vasorelaxation, depending upon the agonists used. Given the recognized role of adenosine in the vasculature, these alterations suggest taurine-mediated modulation of blood flow regulation. PMID- 14746831 TI - Minimally invasive surgery for achalasia: a 10-year experience. AB - Minimally invasive esophagomyotomy for achalasia has become the preferred surgical treatment; the employment of a concomitant fundoplication with the myotomy is controversial. Here we report a retrospective analysis of 53 patients with achalasia treated with laparoscopic Heller myotomy; fundoplication was used in all patients except one, and 48 of the fundoplications were complete (floppy Nissen). There were no deaths or reoperations, and minor complications occurred in three patients. Good-to-excellent long-term results were obtained in 92% of the subjects (median follow-up 3 years). Two cases (4%) of persistent postoperative dysphagia were documented, one of which was treated with dilatation. Postoperative reflux occurred in five patients, four of whom did not receive a complete fundoplication; these patients were well controlled with medical therapy. We suggest that esophageal achalasia may be successfully treated with laparoscopic Heller myotomy and floppy Nissen fundoplication with an acceptable rate of postoperative dysphagia. PMID- 14746830 TI - Barrett's esophagus. PMID- 14746832 TI - Esophageal achalasia: is the herpes simplex virus really innocent? AB - This study was designed to test the hypothesis that mononuclear cells in the myenteric plexus of patients with achalasia may be activated by herpes simplex virus type 1 (HSV-1). Strips of esophageal muscle were obtained from patients with achalasia and multiorgan transplant donors who served as control subjects. After muscle digestion, mononuclear cells were purified through a Percoll gradient and cultured in medium, either alone or containing ultraviolet inactivated HSV-1 or poliovirus (multiplicity of infection 1:1.5). As an indicator of HSV-1-induced lymphocyte activation, we determined T-cell proliferation by means of 3H-thymidine incorporation and interferon gamma release. DNA was extracted from esophageal muscle of achalasia patients and control subjects, and used as a template for PCR analysis using primer pairs specific for HSV-1. Circulating anti-HSV-1 and HSV-2 antibodies were detected by enzyme-linked immunosorbent assay on serum samples. Fifteen patients with naive achalasia and eight control subjects were studied. The prevalence of circulating anti-HSV-1 and HSV-2 antibodies proved similar in the two groups, and no HSV-1 DNA was detected by polyermase chain reaction in the esophageal muscle samples. The proliferative index in mononuclear cells from achalasia patients stimulated with HSV-1 showed a 3.4-fold increase in comparison with control subjects (P<0.01). In addition, a 1.4-fold increase in interferon gamma release after incubation with HSV-1 was observed in cells from achalasia patients but not control subjects. The results of this study indicate that HSV-1-reactive immune cells are present in lower esophageal sphincter muscles of patients with achalasia. We hypothesize that the HSV-1-reactive lymphocytes in lower esophageal sphincter muscles of achalasia patients may contribute to damage of the neurons in the myenteric plexus and lead to the motor dysfunction. PMID- 14746833 TI - Disparity between symptomatic and physiologic outcomes following esophageal lengthening procedures for antireflux surgery. AB - Although esophageal lengthening procedures (Collis gastroplasty) have been recommended as an adjunct to antireflux surgery in patients with shortened esophagus, there are few data on physiologic outcomes in these patients. This study details the long-term outcomes in patients who underwent antireflux surgery with Collis gastroplasty. All patients undergoing esophagogastric fundoplication (EGF) with a Collis gastroplasty for the management of gastroesophageal reflux disease or paraesophageal hernia were identified from a prospectively maintained database. Symptom questionnaires were used during follow-up to assess symptomatic outcomes. Barium esophogram, upper endoscopy with biopsy, and catheterless esophageal acid monitoring (BRAVO system) were recommended for all patients. Patients with abnormal results of physiologic studies underwent further treatment based on a standardized algorithm. Between 1996 and 2002, a total of 68 patients underwent EGF with Collis gastroplasty. Twenty-seven (40%) had a large paraesophageal hernia, and 20 (30%) had undergone a prior EGF. Fifty-six (82%) of the procedures were performed laparoscopically. Mean follow-up time was 30 months, with 10 (15%) patients lost to latest follow-up. Symptomatic outcome data were available for 85% of patients, with significant improvements reported for heartburn (86%), chest pain (90%), dysphagia (89%), and regurgitation (91%). Most patients (84%) were off medications. Physiologic data were completed in 37% of the patients. Of those undergoing physiologic follow-up studies, 17% had recurrent hiatal hernia, and 80% had endoscopically identified esophagitis and pathologic esophageal acid exposure on pH testing. Despite this, 65% of the patients with objectively identified abnormalities reported significant symptomatic improvement compared to their preoperative symptoms. Two patients developed changes associated with Barrett's esophagus that were not present preoperatively. Distal esophageal injury can persist after EGF with Collis gastroplasty, despite significant symptomatic improvements. Appropriate follow-up in these patients requires objective surveillance, which should eventuate in further treatment if esophageal acid is not completely controlled. Although the Collis gastroplasty is conceptually appealing, these results call into question the liberal application of this technique during EGF. PMID- 14746834 TI - Insulin-like growth factor-2 activation of intestinal glutamine transport is mediated by mitogen-activated protein kinases. AB - Insulin-like growth factor-2 (IGF-2) plays a pivotal role in regulating intestinal epithelial metabolism, growth, and proliferation, but its regulatory effects on mucosal cell amino acid transport have not been well studied. The purpose of this in vitro study was to investigate the regulatory mechanisms and intracellular signaling pathways involved in the regulation of IGF-2 on glutamine transport in cultured intestinal cells. Continuous incubation with IGF-2 stimulated glutamine transport activity in cultured IEC-6 cells in a dose- and time-dependent fashion. Prolonged incubation (up to 48 hours) resulted in a 50% increase in transport activity (0.81+/-0.21 nmole/mg protein/min in IGF-2 cells vs. 0.57+/-0.15 nmole/mg protein/min in control cells) and a threefold increase in glutamine transporter ATB(0) mRNA levels. IGF-2 stimulated transport activity by increasing transport maximal capacity (V(max) 4.31+/-0.36 nmole/mg protein/min in IGF-2 cells vs. 2.51+/-0.23 nmole/mg protein/min in control cells) without affecting the transport affinity (K(m) 0.31+/-0.03 mmol/L glutamine in IGF-2 cells vs. 0.28+/-0.03 mmol/L glutamine in control cells). This IGF-2-induced glutamine transport activity was attenuated by actinomycin-D or cycloheximide. The levels of mitogen-activated protein kinases p42/44, MEK1/2, and p38 as well as protein kinase C levels were elevated in IGF-2-treated cells and inhibitors of mitogen-activated protein kinase MEK1 (PD 98059), mitogen-activated protein kinase p38, and protein kinase C (chelerythrine chloride) individually attenuated the IGF-2-induced glutamine transport. These data suggest that IGF-2 stimulates intestinal glutamine uptake in cultured rat intestinal epithelial cells via a mechanism that involves transcription and translation of the transporter. Activation of mitogen-activated protein kinases and protein kinase C cascades are involved in the regulation. This increase in glutamine uptake may occur to support intestinal cell growth and proliferation. PMID- 14746835 TI - Serum fat-soluble vitamin deficiency and abnormal calcium metabolism after malabsorptive bariatric surgery. AB - Weight loss after biliopancreatic diversion or duodenal switch is due to decreased calorie absorption secondary to fat malabsorption. Fat malabsorption may also cause essential fat-soluble vitamin deficiencies, which may have severe clinical consequences and alter calcium metabolism. Serum vitamins A, D, E, and K, zinc, parathyroid hormone, corrected calcium, and alkaline phosphatase levels were measured in a cohort of patients who had previously undergone biliopancreatic diversion. Two bariatric surgery units were involved in the study: New York University School of Medicine (New York, NY), and the Wesley Medical Center (Brisbane, Australia). A total of 170 patients completed the study. The incidence of vitamin A deficiency was 69%, vitamin K deficiency 68%, and vitamin D deficiency 63% by the fourth year after surgery. The incidence of vitamin E and zinc deficiency did not increase with time after surgery. The incidence of hypocalcemia increased from 15% to 48% over the study period with a corresponding increase in serum parathyroid hormone values in 69% of patients in the fourth postoperative year. There is a progressive increase in the incidence and severity of hypovitaminemia A, D, and K with time after biliopancreatic diversion and duodenal switch. Calcium metabolism is affected with an increasing incidence of secondary hyperparathyrodisim and evidence of increased bone resorption in 3% of patients. Long-term nutritional monitoring is necessary after malabsorptive operations for morbid obesity. PMID- 14746836 TI - Predicting the node-negative mesorectum after preoperative chemoradiation for locally advanced rectal carcinoma. AB - Preoperative chemoradiation therapy (CRT) in patients with locally advanced rectal cancer allows for radical surgery with sphincter preservation in many patients. To determine whether patients downsized with preoperative CRT may be potential candidates for local excision, we investigated residual disease patterns after neoadjuvant treatment. A retrospective analysis was carried out of patients with T3 or T4 rectal adenocarcinoma who were treated with neoadjuvant CRT. Clinical and pathologic data were analyzed to (1). determine the response rates to preoperative CRT in the tumor bed and regional nodal basin and (2). identify the incidence of residual disease in the mesorectum in patients downsized to 1cm) but was not helpful for the infiltrating type. PET was also helpful for detecting residual gallbladder carcinoma following cholecystectomy, but was not helpful in patients with carcinomatosis. Although FDG-PET led to a change in management in 30% of patients with cholangiocarcinoma, it must be interpreted with caution in patients with primary sclerosing cholangitis and with stents in place, as well as in those with known granulomatous disease. PMID- 14746841 TI - Specific gene expression and therapy for pancreatic cancer using the cytosine deaminase gene directed by the rat insulin promoter. AB - Suicide gene therapy has been shown to be an effective means of destroying pancreatic cancer cells, but cell-specific delivery of the gene is required to limit host toxicity. The objective of this study is to determine whether the rat insulin promoter (RIP) will permit cell-specific gene delivery and subsequent cell death in human pancreatic cancer cells. The RIP DNA was amplified using polymerase chain reaction (PCR), and the purified fragment was inserted into pCR Blunt II-TOPO plasmid at the SpeI site, which contains the coding sequence of yeast cytosine deaminase (CD). Transfection assays were carried out using both RIP-lacZ and RIP-CD DNA constructs in two human pancreatic cancer cell lines, PANC-1 and MIA PaCa-2. Reporter assays using X-gal staining were performed, and the in vitro cytotoxicity was examined in RIP-CD-transfected cells treated with 5 flucytosine for 5 days. The expression levels of CD protein in the transfected cells were determined 2 days after transfection by Western blot analysis. The expression levels of insulin promoter factor (IPF-1/PDX-1) in these human pancreatic cell lines, as well as in freshly isolated human pancreatic cancer specimens, were determined using in situ immunohistochemistry analysis. After transfection with RIP-lacZ, only PANC-1 cells, but not MIA PaCa-2 cells, were positive for RIP-lacZ expression, indicating that RIP-directed reporter gene expression occurred only in PANC-1 cells. After transfection with RIP-CD and treatment with 5-flucytosine, PANC-1 cells had a significantly increased cell death rate compared with that of MIA PaCa-2 cells, suggesting that RIP-directed suicide gene expression occurred only in PANC-1 cells. Western blot analysis demonstrated that only PANC-1 cells were able to express the CD protein and that significantly increased levels of PDX-1 were found in PANC-1 but not in Mia PaCa 2 cells. In situ immunohistochemical analysis of both cell lines showed that PDX 1 was only expressed in the nuclei of PANC-1 cells and not in MIA PaCa-2 cells. Furthermore, two freshly isolated human pancreatic cancer specimens had significantly increased levels of PDX-1. The RIP is activated in PANC-1 cells, but not in Mia PaCa-2 cells, and the mechanism of activation is via PDX-1. Pancreatic cancer-specific cytotoxicity can be achieved with the use of RIP-CD and 5-flucytosine treatment in vitro. Significantly increased levels of PDX-1 have been found in human pancreatic cancer specimens. These results suggest that RIP could be used for cell-specific suicide gene therapy to target human pancreatic tumors. PMID- 14746842 TI - Clinical application of porcine small intestinal submucosa in the management of infected or potentially contaminated abdominal defects. AB - The repair of abdominal wall defects in potentially contaminated or grossly infected fields presents a difficult clinical problem. Polypropylene mesh is relatively contraindicated in these settings because of the potential for chronic infection. The alternatives to polypropylene include polyglactin mesh, which is not associated with chronic infection but is associated with a 100% recurrence of hernia. The ideal prosthetic for this patient group should be resistant to infection and ensure a low rate of hernia recurrence. We studied the use of small intestinal submucosa, which has been reported to be resistant to infection and incorporates into the fascia over 3 to 6 months, in 20 patients with ventral or inguinal hernias (18 ventral, 2 inguinal hernia) in the setting of bacterial contamination. The early postoperative complication rate was 50%. One patient with fasciitis had degradation of the small intestinal submucosa and loss of the bioprosthesis within 7 days. Other early complications included seroma (n=2), ileus (n=1), and wound infection (n=8). No patient experienced chronic infection. Mean follow up was 15.7 months and the rate of recurrence documented by CT or physical examination was 30%. We concluded the following: (1). small intestinal submucosa is an effective alternative bioprosthesis in the management of ventral/inguinal hernia when there is associated bacterial contamination; (2). human vs. pig immune response has not been seen in this patient population; (3). early graft failure due to overwhelming fascial infection was noted in one patient and may be a limitation of this technology in a minority of patients; and (4). early hernia recurrence is relatively low but long-term follow-up has not been completed. PMID- 14746843 TI - Octreotide improves reperfusion-induced oxidative injury in acute abdominal hypertension in rats. AB - Ischemia/reperfusion injury plays an important role in the pathogenesis of abdominal compartment syndrome, which is characterized by increased intra abdominal pressure. The aim of this study was to investigate whether octreotide, a synthetic somatostatin analogue, improves the reperfusion injury after decompression of acute abdominal hypertension. This study was carried out in Wistar albino rats. With the rats under anesthesia, an arterial catheter was inserted intraperioneally and with the use of an aneroid manometer connected to the catheter, intra-abdominal pressure was kept at 20 mm Hg (ischemia group) for 1 hour. In the ischemia/reperfusion group, pressure applied for 1 hour was decompressed and a 1-hour reperfusion period was allowed. In another ischemia/reperfusion group, octreotide was administered (50 microg/kg intraperitoneally) immediately before the decompression of intra-abdominal pressure. At the end of the experiment, liver and intestinal tissues were taken and malondialdehyde (an index of lipid peroxidation) and glutathione (a key to antioxidant) levels and myeloperoxidase (an index of tissue neutrophil infiltration) activity were estimated. The results demonstrated that tissue levels of malondialdehyde and myeloperoxidase activity were elevated, whereas glutathione levels were reduced in both the ischemia and ischemia/reperfusion groups. Octreotide treatment reversed these oxidant responses. In conclusion, increased intra-abdominal pressure causes oxidative organ damage and octreotide, by controlling the reperfusion of abdominal organs and inhibiting neutrophil infiltration, could improve the reperfusion-induced oxidative damage. Therefore its therapeutic role as a "reperfusion injury-limiting" agent must be further elucidated in intra-aortic pressure-induced abdominal organ injury. PMID- 14746844 TI - Laparoscopic splenectomy reverses thrombocytopenia in patients with hepatitis C cirrhosis and portal hypertension. AB - Pegylated-interferon (IFN) plus ribavirin remains the most effective therapeutic regimen for patients with chronic hepatitis C infection. Thrombocytopenia is a common side effect of this treatment, often leading to discontinuation of a potentially curative therapy. We sought to determine the safety and efficacy of laparoscopic splenectomy in correcting thrombocytopenia, thus allowing completion of IFN therapy. Data were collected prospectively from September 2000 to May 2003 on all patients undergoing laparoscopic splenectomy for thrombocytopenia associated with IFN therapy and/or hepatitis C cirrhosis with portal hypertension. Demographic data, model of end-stage liver disease (MELD) score, platelet count, operative time, blood loss, spleen weight, complications, length of stay, and follow-up time were calculated. Eleven patients (7 men, 4 women) underwent laparoscopic splenectomy; their mean age was 45.4 years (range 27 to 55 years) and mean body mass index was 27 kg/m(2) (range 21 to 44 kg/m(2)). All patients were Child's class A, with a mean preoperative MELD score of 9.1 (range 6 to 11). Mean operative time was 189 minutes (range 70 to 245 minutes), and blood loss averaged 141 ml (range 10 to 600 ml). A hand-assisted laparoscopic technique was used in four cases. Six patients received empiric intraoperative platelet administration. None required transfusion with packed red cells. Splenic weight averaged 1043 g (range 245 to 1650 g). Average length of stay was 2.6 days (range 1 to 6 days). Four patients had the following minor postoperative complications: self-limited atrial fibrillation (n=1), trocar site cellulitis (n=1), and atelectasis (n=2). There have been no major complications over an average follow-up of 11 months (range 1 to 18 months). Mean postoperative MELD score was 8.3 (range 6 to 10). Platelet counts improved from a preoperative mean of 55000/ul (16000 to 88000/microl) to 439000/microl (200000 to 710000/microl) postoperatively and have remained above 100000/microl (104000 to 397000/microl) during subsequent pegylated-IFN therapy. Three patients have completed a full course of IFN therapy and have obtained a sustained virologic response. Treatment is ongoing in the remaining patients. Laparoscopic splenectomy is safe in the setting of portal hypertension and thrombocytopenia associated with chronic hepatitis C infection. It can be performed with little blood loss, no need for red cell transfusion, and minimal perioperative morbidity. Laparoscopic splenectomy appears to effectively reverse thrombocytopenia and may allow these patients to safely complete IFN therapy. PMID- 14746845 TI - Bovine pericardium buttress limits recanalization of the uncut Roux-en-Y in a porcine model. AB - In contrast to the traditional Roux-en-Y reconstruction, an uncut Roux-en-Y provides biliopancreatic diversion and may preserve myoelectric continuity. Previous iterations of the uncut Roux have been plagued by recanalization of the uncut staple line in the afferent small bowel. Our aim was to determine if bovine pericardium buttress prevents recanalization of the stapled small bowel partition in a porcine model. Sixteen female pigs ( approximately 30 kg) underwent a side to-side stapled jejunojejunostomy, 20 cm distal to the ligament of Treitz, with placement of a nondivided stapled partition with a single row of 2.5 mm width staples in the intervening jejunal loop. Nine animals in the experimental group had a bovine pericardium buttressed staple line (5 permanent, 4 absorbable), whereas seven animals in the control group had a nonbuttressed staple line. At 6 or 12 weeks, necropsy was performed and the primary outcome, staple line recanalization, was assessed grossly and histologically. Statistical analysis was performed by means of the chi-square test. There were no major complications and all animals gained weight. Overall, eight of nine bovine pericardium buttressed staple lines were grossly and histologically intact at necropsy, whereas all nonbuttressed uncut staple lines had recanalized completely (P<0.05). At 6 weeks, both permanent (N=4) and absorbable (N=3) buttress preparations prevented recanalization. At 12 weeks the permanent buttress remained closed (N=1), but the absorbable buttress had allowed partial recanalization (N=1). The use of bovine pericardium buttress will prevent small bowel recanalization of uncut small bowel staple lines at early follow-up. Pilot data at intermediate follow-up suggest permanent buttress is more durable than absorbable buttress. These results warrant investigation of bovine pericardium for intestinal applications in humans. PMID- 14746846 TI - Laparoscopic Nissen fundoplication for treating reflux in lung transplant recipients. AB - Gastroesophageal reflux disease may contribute to pulmonary injury and the development of bronchiolitis obliterans syndrome in lung transplant patients. As a result, such individuals are increasingly likely to undergo corrective gastrointestinal surgery. The present study collected outcome data for 28 lung transplant patients with documented reflux who underwent an uncomplicated laparoscopic Nissen fundoplication at our institution. The results were compared to data from 63 nontransplant reflux patients who had undergone the procedure over the same time period. All Nissen fundoplications were conducted by the same surgeon. There were no intraoperative or perioperative deaths in either patient group. Operative parameters did not differ but the postoperative hospital stay was significantly greater for the lung transplant patients (P<0.05). Seven transplant patients (25%) were readmitted within 30 days compared to two readmissions (3.2%) in the reflux group. Five transplant patients (17.9%) have died, all from pulmonary complications; on average, death occurred 15.5 months after the Nissen surgery. There have been no deaths in the reflux group. These data indicate that laparoscopic Nissen fundoplication can be performed on lung transplant recipients to treat reflux. The average hospital stay is longer and there are more frequent readmissions in this population, but this does not appear to be due to any Nissen-related morbidity. PMID- 14746847 TI - Behavioural and social science research in cancer: time for action. PMID- 14746848 TI - Research in the behavioural and social sciences to improve cancer control and care: a strategy for development. A report of an expert group. AB - The need for a better co-ordinated interdisciplinary approach to cancer is widely recognised. An analysis of strengths and weaknesses has emphasised the importance of a better understanding of the behavioural and social factors which determine the success of preventative and screening programmes and those which will enhance the quality of care and support available to patients and their families. The European Commission provided funds to establish a consultation to formulate a strategy for the development of research in the behavioural, social and related sciences relevant to cancer. The key objectives were to inform research organisations of the steps necessary to enhance research capacity in these areas and identify the most productive directions for research in the forseeable future. The expert group identified 11 areas in which research might be expected to improve cancer control and treatment. PMID- 14746849 TI - Period analysis for 'up-to-date' cancer survival data: theory, empirical evaluation, computational realisation and applications. AB - Long-term survival rates are the most commonly used outcome measures for patients with cancer. However, traditional long-term survival statistics, which are derived by cohort-based types of analysis, essentially reflect the survival expectations of patients diagnosed many years ago. They are therefore often severely outdated at the time they become available. A couple of years ago, a new method of survival analysis, denoted period analysis, has been introduced to derive more 'up-to-date' estimates of long-term survival rates. We give a comprehensive review of the new methodology, its statistical background, empirical evaluation, computational realisation and applications. We conclude that period analysis is a powerful tool to provide more 'up-to-date' cancer survival rates. More widespread use by cancer registries should help to increase the use of cancer survival statistics for patients, clinicians, and public health authorities. PMID- 14746850 TI - Wide metastatic spreading in infiltrating lobular carcinoma of the breast. AB - The aim of this study was to determine whether the metastatic potential of breast cancer could be related to phenotypic characteristics of the tumour. Therefore, we compared the metastatic patterns of invasive lobular (ILC) and ductal (IDC) carcinomas. In ILC, we also analysed this pattern according to the histological subtype of the primary and the E-cadherin (EC) expression level. Metastatic ILC cases (n=96) were retrospectively analysed and classified into classical, alveolar, solid, tubulo-lobular, signet ring cells or pleomorphic subtypes. Anatomical distribution of metastases was detailed for every patient and compared with that registered for IDC (n=2749). Immunostaining of EC (HECD1 antibody) was performed in 82 cases. Histologically, 78 of the 96 cases (81%) corresponded to classical ILC. The pleomorphic subtype was observed in 14 cases (15%), a rate that was higher than that expected. Others corresponded to alveolar (2 cases), signet ring cell (1 case) and solid (1 case) subtypes. EC was undetectable in 72/82 cases (88%). The rate of multiple metastases was higher in ILC (25.0%) than in IDC (15.8%) (P=0.016). Metastases were found more frequently in ILC than in IDC in the bone (P=0.02) and/or in various other sites (peritoneum, ovary, digestive tract, skin em leader ) (P<0.001). In ILC, no significant link was found between the localisation(s) of metastases, the histological subtype and the EC status in the primary. In conclusion, in breast carcinomas, the frequency of multiple metastasis was found to be higher in ILC than IDC. This fact may be related to the phenotypic trait of discohesive small cells which characterises ILC. EC loss, observed in most cases of ILC, may result in alterations in cell cell adhesion and a preferential growth at metastatic sites. A high rate of pleomorphic tumours was observed in the group of metastatic ILC, but the pattern of metastatic site(s) was not related to the histological subtype of the primary. PMID- 14746851 TI - The poor responsiveness of infiltrating lobular breast carcinomas to neoadjuvant chemotherapy can be explained by their biological profile. AB - The aim of this study was to determine the chemosensitivity of infiltrating lobular breast carcinoma (ILC) in comparison with infiltrating ductal carcinoma (IDC). Between 1987 and 1995, 457 patients with invasive T2>3 cm-T4 breast carcinomas were treated with primary chemotherapy (CT), surgery, radiation therapy. Clinical response, the possibility of breast preservation, pathological response and survival were evaluated according to the histological type. In order to evaluate the biological differences between ILC and IDC patients and their implication with regard to tumour chemosensitivity, additional immunohistochemical stainings (oestrogen receptor (ER), Bcl2, p53, c-erbB-2 and Ki67) were performed on 129 pretherapeutical specimens. 38 (8.3%) ILC were diagnosed by core needle biopsy before CT. ILC was an independent predictor of a poor clinical response (P=0.02) and ineligibility for breast-conserving surgery after neoadjuvant chemotherapy (P=0.03). Histological and biological factors predicting a poor response to CT (histological grade, ER, Ki67 and p53 status) were more frequent in ILC than in IDC patients. After a median follow-up of 98 months (range: 3-166), the low chemosensitivity of ILC did not result in a survival disadvantage. Our results demonstrate that ILC achieved a lower response to CT than IDC because of their immunohistochemical profile. Preoperative CT did not allow a high rate of conservative treatment for ILC and therefore the use of neoadjuvant CT for ILC patients should be questioned. PMID- 14746852 TI - Weekly paclitaxel as first-line chemotherapy for elderly patients with metastatic breast cancer. A multicentre phase II trial. AB - Paclitaxel is a cytotoxic agent with proven antitumour activity in metastatic breast cancer. Weekly administration of paclitaxel has demonstrated sustained efficacy together with a more favourable toxicity profile (e.g. less myelotoxicity) than the 3-weekly administration. This study evaluates the activity and toxicity of weekly paclitaxel (Taxol(R)) as first-line chemotherapy in elderly patients (>70 years of age) with hormone-refractory metastatic breast cancer. Patients with metastatic breast cancer received 80 mg/m(2) paclitaxel administered weekly on days 1, 8 and 15 of a 28-day cycle. Additional cycles were given until disease progression, or unacceptable toxicity. A dose increase to 90 mg/m(2) was allowed in the absence of toxicity. 26 Patients received a total of 101 cycles (median 4, range 1-11). 22 patients completed at least two cycles (six administrations). In 23 patients who were evaluable for response, there were 10 partial responses (38%), 9 patients with stable disease (35%), while 4 patients had disease progression (15%). The median duration of response was 194 days (>6 months). Overall treatment was relatively well tolerated, but 8 patients (32%) had to prematurely discontinue treatment because of fatigue. Neuropathy >grade 1 was noted only after five or more cycles in 4 patients. Weekly paclitaxel at this dose and schedule is an effective treatment regimen in the elderly patient with metastatic breast cancer, and is feasible, but yields relevant fatigue in a subset of patients. PMID- 14746853 TI - Selective immunohistochemical staining shows significant prognostic influence of lymphatic and blood vessels in patients with malignant melanoma. AB - Few data on the influence of lymphatic microvessel density (LMVD) on survival in patients with melanoma are available. The aim of this study was to assess LMVD and blood microvessel density (MVD) in tissue samples from 120 patients with melanoma. LMVD was stained with an antibody staining for podoplanin, and blood MVD was assessed by CD31 (PECAM-1)-immunostaining. Survival was determined using univariate and multivariate analysis. A significant association between a high CD31 MVD (but not LMVD) and the presence of lymph node metastases (P=0.007) was observed. Patients with a high LMVD had a significant shorter overall (OS) (P=0.0436) and disease-free survival (DFS) (P=0.0249) in univariate analysis. The survival analysis showed CD31 MVD was a strong prognostic factor for OS and DFS in both uni-and multivariate analyses. Our results demonstrate LMVD as a prognostic factor in malignant melanoma, although its prognostic relevance is much smaller compared with blood MVD. PMID- 14746854 TI - Preoperative psychological reactions and quality of life among women with an increased risk of breast cancer who are considering a prophylactic mastectomy. AB - A consecutive sample of 56 women with a familial risk for breast cancer who were considering a prophylactic mastectomy (PM) completed questionnaires preoperatively concerning risk perception, expectations with regard to surgery, anxiety and depressive symptoms (the Hospital Anxiety and Depression Scale (HAD) scale) and quality of life (The Swedish SF-36 Health Survey). 16 had had a previous breast cancer (Group BC) and 40 had not (Group R). They were compared with normative data from an age-matched random sample of the Swedish population and with a reference sample of women with breast cancer. Most women estimated their breast cancer risk accurately. No statistically significant differences were found between Group BC and the normative sample on the HAD scale and SF-36, but Group R reported better physical functioning, emotional role functioning and mental health than the reference sample with breast cancer. Group BC scored closer to them than to the normative sample. Levels of emotional problems and quality of life were comparable to normative values among women considering PM. All women in the present study had previous genetic counselling and our results suggest that their interest in PM was not due to an overestimation of their personal risk. PMID- 14746855 TI - 'State of the art' of radical hysterectomy; current practice in European oncology centres. AB - Quality control of medical performance requires adequate 'state-of-the-art' data and this is currently not uniformly defined for radical hysterectomy. We have used data from a randomised multicentre clinical trial examining the clinical significance of surgical drains following radical hysterectomy (European Organisation for Research and Treatment of Cancer (EORTC)-55962). Although the study was not designed to analyse the quality of the surgical procedure per se, surgical data during and after the operation were carefully noted. A total of 234 patients from 12 European institutes were included in the study. We reported on the clinical and surgical characteristics, the radicality of surgery and short- and long-term complications of radical hysterectomy: median duration of surgery: 240 min; median number of nodes removed: 26; lymph node metastases: 22%; post operative mortality: <1%; urinary tract infection: 42%; deep venous thrombosis: 3%; fistula: 2%. The data from our study provides an honest and realistic picture of the current practice of radical hysterectomy among European oncology centres and may be considered as the 'standard of care' in this part of the world. PMID- 14746856 TI - Central nervous system progression among patients with metastatic breast cancer responding to trastuzumab treatment. AB - Central nervous system (CNS) metastases from breast cancer are common and can present as the first or solitary site of disease progression. The CNS has been reported to act as a sanctuary site that denies access to many chemotherapeutic agents. We present here, a series of 10 metastatic breast cancer patients who developed CNS metastases after an initial response to trastuzumab treatment. Forty one patients with metastatic HER2-overexpressing breast cancer, without evidence of CNS involvement prior to the initiation of trastuzumab treatment, were followed during trastuzumab treatment. A neurological evaluation was performed in those patients who developed neurological signs or symptoms during the course of treatment. The clinical course and pattern of CNS involvement in these patients are discussed. Thirty two patients (78%) showed an initial response to trastuzumab treatment. Ten (31%) of the responding patients developed either isolated CNS relapse or concurrent CNS and systemic progression at a median of 43 weeks after the initiation of trastuzumab treatment. Trastuzumab as a single agent was continued following control of brain symptoms in three patients, two showed signs of systemic disease progression at 11 and 15 weeks following the diagnosis of CNS metastases, respectively. In two other patients, trastuzumab in combination with weekly chemotherapy was continued for more than 20 weeks after CNS relapse without evidence of disease progression. The incidence of CNS involvement in our group of patients was higher than expected. With more successful and prolonged systemic anti-tumour effects achieved by novel drug combinations, the risk of developing CNS metastases might be even greater. Evaluation of prophylactic cranial irradiation strategies might be studied for high-risk patients. PMID- 14746857 TI - Second malignancies and Richter's syndrome in patients with chronic lymphocytic leukaemia treated with cladribine. AB - The increased frequency of second malignancies in chronic lymphocytic leukaemia (CLL) is well known. Moreover, antineoplastic therapy additionally increases the risk of secondary cancers. In this study, we analysed whether treatment with cladribine (2-chlorodeoxyadenosine, 2-CdA) during the course of CLL had an impact on the subsequent occurrence of either secondary solid tumours or Richter's syndrome. There were 1487 eligible patients, 251 treated with 2-CdA alone, 913 treated with alkylating agents (AA)-based regimens alone and 323 treated with both 2-CdA and AA. Median time from the start of CLL treatment to the diagnosis of secondary malignancy was 1.9 years (0.5-5.1 years) for the 2-CdA group, 1.8 years (0.3-7.9 years) for the AA group and 3.9 years (0.3-8.4 years) for the 2 CdA+AA group. A total of 68 malignancies were reported in 65 patients. Ten events were non-melanotic skin cancers and were excluded from the analysis, leaving 58 events in 58 patients. In the group of patients treated with 2-CdA alone, there were 15 (6.0%) cases, in the group of patients treated with AA alone there were 26 (2.8%) cases, and in the group treated with 2-CdA+AA there were 17 (5.3%) cases of secondary malignancies. The differences between the frequency of secondary malignancies in the 2-CdA and 2-CdA+AA versus AA alone groups were not significant (P=0.05 and P=0.06, respectively). Only lung cancers occurred significantly more frequently in the 2-CdA (2.8%) and 2-CdA+AA (2.2%) treated groups compared with the AA patients (0.3%) (P<0.001 and P<0.01, respectively). In conclusion, 2-CdA in CLL patients does not seem to increase the risk of secondary malignancies except for lung cancers. However, further studies are necessary to establish the real risk of lung cancer in CLL patients treated with 2-CdA. PMID- 14746858 TI - Final results of the EORTC 18871/DKG 80-1 randomised phase III trial. rIFN alpha2b versus rIFN-gamma versus ISCADOR M versus observation after surgery in melanoma patients with either high-risk primary (thickness >3 mm) or regional lymph node metastasis. AB - Between 1988 and 1996, the European Organisation for Research and Treatment of Cancer Melanoma Group (EORTC-MG) performed a prospective, randomised phase III adjuvant trial to evaluate the efficacy and toxicity of low dose recombinant interferon-alpha 2 b (rIFN-alpha2b) (1 MU) or recombinant interferon gamma (rIFN gamma), (0.2 mg) both given subcutaneously (s.c.), every other day (qod), for 12 months in comparison with an untreated control group. The German Cancer Society (DKG) added a fourth arm with Iscador M, a popular mistletoe extract. High-risk stage II patients (thickness >3 mm) and stage III patients (positive lymph nodes) without distant metastasis were randomised and followed until their first progression or death. An intention-to-treat analysis was performed. From 1988 to 1996, a total of 830 patients were randomised: 423 in the three-arm EORTC 18871 trial and 407 patients in the four-arm DKG 80-1 trial. The median follow-up was 8.2 years and a total of 537 relapses and 475 deaths were reported. At 8 years, the disease-free interval (DFI) rate was 32.4% and the overall survival (OS) rate was 40.0%. In terms of the DFI, the hazard ratio estimates (95% Confidence Intervals (CI)) were: 1.04 (0.84, 1.30) for the comparison of rIFN-alpha2b versus control, 0.96 (0.77, 1.20) for rIFN-gamma versus control, and 1.32 (0.93, 1.87) for Iscador M versus control. In terms of OS, the corresponding estimates (95% CI) for the 3 treatment comparisons were: for IFN-alpha2b 0.96 (0.76, 1.21), for rIFN-gamma 0.87 (0.69, 1.10) and for Iscador M 1.21 (0.84, 1.75), respectively. The results show no clinical benefit for adjuvant treatment with low dose rIFN alpha2b or rIFN-gamma or with Iscador M in high-risk melanoma patients. PMID- 14746859 TI - The oral NK(1) antagonist, aprepitant, given with standard antiemetics provides protection against nausea and vomiting over multiple cycles of cisplatin-based chemotherapy: a combined analysis of two randomised, placebo-controlled phase III clinical trials. AB - In early clinical trials, the NK(1) receptor antagonist, aprepitant (EMEND(R)) was shown to improve the protection provided by the best available therapy (hereafter referred to as 'standard therapy': a 5-HT(3) receptor antagonist and dexamethasone) against chemotherapy-induced nausea and vomiting over multiple cycles of cisplatin-based chemotherapy. To further study the sustainment of antiemetic efficacy of aprepitant plus standard therapy over more than one cycle of chemotherapy, we examined combined data from the multiple cycles extensions of two phase III clinical trials of oral aprepitant plus standard therapy for the prevention of chemotherapy-induced nausea and vomiting. Data were pooled from two multicentre, randomised, double-blind, placebo-controlled studies with identical design and treatment regimens. Cancer patients receiving a first cycle of cisplatin-based (>or=70 mg/m(2)) chemotherapy were randomised to one of two treatment groups as follows: the standard therapy group received ondansetron 32 mg intravenously (i.v.) and dexamethasone 20 mg on day 1 and dexamethasone 8 mg twice daily (b.i.d.) on days 2-4. The aprepitant group received aprepitant 125 mg, ondansetron 32 mg i.v., and dexamethasone 12 mg on day 1, aprepitant 80 mg and dexamethasone 8 mg on days 2-3, and dexamethasone 8 mg on day 4. Patients had the option to receive the same blinded treatment for up to five additional cycles. The analysis used a combined exploratory endpoint of no emesis and no significant nausea (i.e. nausea which interfered with a patient's normal activities) over the 5 days following cisplatin, for up to six cycles of chemotherapy. A cumulative probabilities approach incorporating a model for transitional probabilities was used to analyse the data. Tolerability was assessed by reported adverse events and physical and laboratory assessments. Baseline characteristics, reasons for discontinuation, and drop-out rates were similar between groups. In every cycle, the estimated probabilities (rates) of no emesis and no significant nausea were significantly higher (P<0.006) in the aprepitant group: in the first cycle, rates were 61% in the aprepitant group (N=516) and 46% in the standard therapy group (N=522), and thereafter, rates for the aprepitant regimen remained higher throughout (59% (N=89) versus 40% (N=78) for the standard therapy, by cycle 6). Repeated dosing with aprepitant over multiple cycles was generally well tolerated. Compared with patients who received standard therapy alone (a 5-HT(3) antagonist plus dexamethasone), those who received aprepitant in addition to standard therapy had consistently better antiemetic protection that was well maintained over multiple cycles of highly emetogenic chemotherapy PMID- 14746860 TI - Risk-adapted treatment for childhood hepatoblastoma. final report of the second study of the International Society of Paediatric Oncology--SIOPEL 2. AB - SIOPEL 2 was a pilot study designed to test the efficacy and toxicity of two chemotherapy (CT) regimens, one for patients with hepatoblastoma (HB) confined to the liver and involving no more than three hepatic sectors ('standard-risk (SR) HB'), and one for those with HB extending into all four sectors and/or with lung metastases or intra-abdominal extra hepatic spread 'high-risk (HR) HB'. SR-HB patients were treated with four courses of cisplatin (CDDP), at a dose of 80 mg/m(2) every 14 days, delayed surgery, and then two more similar CDDP courses. HR-HB patients were given CDDP alternating every 14 days with carboplatin (CARBO), 500 mg/m(2), and doxorubicin (DOXO), 60 mg/m(2). Two courses of CARBO/DOXO and one of CDDP were given postoperatively. Between October 1995 and May 1998, 77 SR-HB (10 of whom were actually treated with the HR protocol) and 58 HR-HB patients were registered and all 135 could be evaluated. Response rates for the entire SR-HB and HR-HB groups were 90% (95% CI 80-96%) and 78% (95% CI 65 87%), and resection rates were 97% (95% CI 87-99%) and 67% (95% CI 54-79%) including several children undergoing liver transplantation. For SR-HB patients, 3-year overall and progression-free survivals were 91% (+/-7%) and 89% (+/-7%) and for the HR-HB group 53% (+/-13%) and 48% (+/-13%), respectively. The short term toxicity of these regimens was acceptable, with no toxic deaths. A treatment strategy based on CDDP monotherapy and surgery thus appears effective in SR-HB but, despite CT intensification, only half of the HR-HB patients are long-term survivors. For SR-HB patients, the efficacy of CDDP monotherapy and the CDDP/DOXO ('PLADO') combination are now being compared in a prospective randomised trial (SIOPEL 3). PMID- 14746861 TI - One in 10 ovarian cancer patients carry germ line BRCA1 or BRCA2 mutations: results of a prospective study in Southern Sweden. AB - At least 10% of all ovarian cancers are estimated to have a hereditary background. Hereditary breast-ovarian cancer (HBOC) due to mutations in the BRCA genes is a major cause of hereditary ovarian cancer, although its frequency and relationship to age and family history in unselected series of ovarian cancers is not completely known. We report here the results of a full mutational screening analysis for germ line BRCA1 and BRCA2 mutations in 161 patients with invasive epithelial ovarian carcinomas. Age at diagnosis ranged from 22 to 82 years (mean 59 years). Deleterious (frame-shift, nonsense and missense) mutations were detected in 13/161 (8%) of the patients and affected BRCA1 in 12 cases and BRCA2 in one case. Four additional missense variants (one in BRCA1 and three in BRCA2) with a possible association with an increased risk ovarian cancer were revealed, resulting in a total frequency of BRCA gene alterations of 17/161 (11%). The 13 patients with deleterious mutations had a mean age of 57 years (range 41-76 years) and only three of these patients were below 50 years of age. A family history of at least one breast cancer and/or ovarian cancer was reported in all but 1 of the patients with BRCA mutations compared with only 24% of patients without mutations. Our findings in this prospective study confirm approximately 1 in 10 patients with ovarian cancer carry a germ line BRCA gene mutation associated with HBOC, and also indicate that a large number of these patients are over 50 years of age at diagnosis. PMID- 14746862 TI - Exposure to the sun and sunbeds and the risk of cutaneous melanoma in the UK: a case-control study. AB - Migration, latitude and case-control studies have clearly established a link between melanoma and sun exposure. This case-control study of melanoma was set up to examine the role of sun exposure and sunbeds in the pathogenesis of melanoma in the United Kingdom (UK), a country with low levels of ultraviolet radiation. The study included 413 cases and 416 controls. More than 10 severe sunburns compared with less than 10 sunburns was associated with an Odds Ratio (OR) of 1.98 (95% Confidence Interval (CI) 1.02-3.86) (P=0.04) when adjusted for age, gender and skin type. Sunburns before the age of 15 years were not associated with melanoma once adjustments for age, gender and skin were made. 31% of women and 16% of the men had used sunbeds. Sunbed users were younger than non-users (40 years versus 51 years, P<0.0001). Ever use of sunbeds gave an adjusted OR of 1.19 (95% CI 0.84-1.68) (P=0.33). The risk of melanoma did not increase with increasing hours or years of sunbed exposure. The risk associated with sunbed use was only significant for young individuals with fair skin for whom there was a significant OR of 2.66 (95% CI 1.66-6.09) (P=0.02) after adjustment for the sun exposure variables. Outdoor occupation and residence in hot countries were not associated with an increased risk of melanoma. The only significant associations in this study were with 10 or more sunburns and the use of a sunbed in young subjects with fair skin. Sunbed use is now becoming more prevalent in Caucasian populations and the results of this study suggest that sunbed usage may moderately affect individuals with sun-sensitive skin types. However, the magnitude of melanoma risk in association with natural and artificial sun exposure is small compared with phenotypic risk factors such as skin type and naevus counts. However, it is possible that the mean lag time of 7 years between exposure to sunbed and melanoma in this study may have led to an under-estimation of the long-term melanoma risk. PMID- 14746863 TI - Antagonists of growth hormone-releasing hormone (GH-RH) enhance tumour growth inhibition induced by androgen deprivation in human MDA-Pca-2b prostate cancers. AB - In the present study, we investigated whether the growth hormone-releasing hormone (GH-RH) antagonist JV-1-38 could enhance the effects of androgen deprivation produced by the anti-androgen Flutamide and luteinising hormone releasing hormone (LH-RH) agonist Decapeptyl in an experimental model of human androgen-sensitive MDA PCa 2b prostate carcinoma implanted subcutaneously (s.c.) into nude mice. We also evaluated by reverse transcriptase-polymerase chain reaction (RT-PCR) the effects of combined treatment on the mRNA expression for prostate-specific antigen (PSA) and measured serum PSA levels. In experiment 1, GH-RH antagonist JV-1-38 greatly inhibited tumour growth in combination with Decapeptyl, but was ineffective when given alone. Thus, combined therapy with JV 1-38 at 20 microg/day and Decapeptyl microcapsules releasing 12.5 microg/day for 29 days inhibited significantly (P<0.01) MDA PCa 2b tumour growth by 65%, compared with controls. Combined treatment also significantly (P<0.05) decreased serum PSA levels by 52% and reduced tumour weight by 54% vs. controls. In experiment 2, GH-RH antagonist JV-1-38 at 20 microg/day likewise showed powerful growth inhibitory effects when combined with Flutamide (25 mg/kg/day) for 21 days. Combined treatment with JV-1-38 and slow-release pellets of Flutamide significantly (P<0.001) inhibited tumour growth by 61% versus controls, and was significantly (P<0.05) more effective than Flutamide or JV-1-38 alone. Combination therapy also reduced significantly (P<0.001) tumour weight and serum PSA levels by 59 and 47%, respectively. The mRNA expression for PSA in MDA PCa 2b tumours was not changed by JV-1-38, Decapeptyl and Flutamide alone or in their respective combinations. Our findings suggest that GH-RH antagonists could enhance the tumour inhibitory effects of androgen deprivation for the primary therapy of patients with advanced prostate carcinoma. PMID- 14746864 TI - Alkaline single-cell gel electrophoresis (comet assay): a simple technique to show genomic instability in sporadic breast cancer. AB - The alkaline comet assay was used to study the genomic instability of lymphocytes derived from untreated sporadic breast cancer patients (50 cases), and also following their in vitro irradiation up to 5 Gy. We compared the results (mean tail moment (MTM)) with a control population of 25 patients and with breast cancer patients who had been 'cured' of their disease, with a follow-up of 10 years or more (25 cases). At the basal level, 77.5% (P<0.01) of the untreated patients and 73.7% (P<0.05) of the 'cured' women had values higher than the basal cut-off level of 5.3, compared with only 44% of the controls. After in vitro irradiation, 83% of the untreated patients were above the cut-off value of 10.8 at the 5-Gy dose compared with only 48% of the controls (P<0.01). These results support the hypothesis that women affected by sporadic breast cancer have a constitutional genomic instability. The assessment of the prognostic value of this test could be of interest, particularly in women without axillary nodal involvement. PMID- 14746865 TI - Detection of cell-free nucleic acids in bronchial lavage fluid supernatants from patients with lung cancer. AB - The aim of this study was to determine whether nucleic acids are detectable in cell-free bronchial lavage supernatants, and whether it is possible to find alterations in this DNA and RNA of genes known to be present in lung tumour cells. DNA was isolated from cell-free lavage supernatants from 30 and RNA from 25 lung cancer patients. The DNA was examined for microsatellite alterations (MA) and the RNA analysed for the expression of seven tumour-associated genes. Intact DNA and mRNA could be isolated from all cell-free bronchial lavage supernatants. MA were found in lavage supernatants of 12/30 patients and in lavage cells of 6/30 patients. Altogether alterations were found in 14/30 patients. Analyses of tumour-associated gene expression showed positive results, with at least one marker in the lavage supernatants of all 25 patients. Thus, we could demonstrate, for the first time, that it is possible to isolate intact DNA and RNA from cell free bronchial lavage supernatants. Their quantity and quality is sufficient for further amplification by polymerase chain reaction (PCR)/reverse transcriptase (RT)-PCR. Altogether, tumour-associated changes were detected in DNA samples from 47% of the patients and in RNA samples from all of the patients analysed. PMID- 14746866 TI - Heterogeneity of immunostaining for tumour markers in non-small cell lung carcinoma. AB - Lung carcinoma is a leading cause of death. However, there are few indicators that can aid in prediction and prognosis. Many tumour markers are available, but their reliability is questionable. For example, Ki-67 expression has been associated with increased as well as decreased survival or with no clinical significance. The varying results have been attributed to the methodology, relative intensity of staining, variety of marking and statistical methods. To determine whether differential expression of markers within tumours may be a contributory factor to this lack of agreement, we used two marking methods to evaluate the level of expression of Ki-67, p53 and bcl-2, in addition to the apoptotic index, in serial sections of non-small cell carcinoma. All stains exhibited a degree of heterogeneity. This small study highlights the importance of standardisation of marking methods and interpretation of results if tumour markers are to be used as predictive or prognostic factors. PMID- 14746867 TI - Temporal dynamics and magnitude of the blood flow response at the optic disk in normal subjects during functional retinal flicker-stimulation. AB - Near-infrared laser Doppler flowmetry was applied in 15 normal volunteers to record the time course and magnitude of changes in the velocity (Vel), volume (Vol) and flow (F) of blood and tissue reflectance (R) at the optic disk in response to 40 and 50 s of increased retinal neural activity. This activity was evoked by diffuse luminance flicker of the retinal posterior pole. After 20 s of flicker, the group averages of Vel, Vol, and F were significantly higher than at baseline (pre-flicker) by 12, 24 and 38%. Time constants of the increases in Vel, Vol, and F were 3.4, 12.7 and 9.1 s, respectively. The group average change in R of 1% was not significant. However, in one subject, 15 recordings from the same site of the optic disk showed a significant increase in R of 8%, with a time course similar to that of Vol. Our findings show that, in the human optic nerve, a white matter tissue, the temporal dynamics and magnitude of the response of blood flow to an increase in retinal neural activity are similar to those reported for brain gray matter. Furthermore, although the R-response could be due, in part, to changes in blood volume, other factors, such as activity-evoked tissue scattering changes, may also affect this response. PMID- 14746868 TI - An electrophysiological investigation of memory encoding, depth of processing, and word frequency in humans. AB - Memory encoding can be studied by monitoring brain activity correlated with subsequent remembering. To understand brain potentials associated with encoding, we compared multiple factors known to affect encoding. Depth of processing was manipulated by requiring subjects to detect animal names (deep encoding) or boldface (shallow encoding) in a series of Chinese words. Recognition was more accurate with deep than shallow encoding, and for low- compared to high-frequency words. Potentials were generally more positive for subsequently recognized versus forgotten words; for deep compared to shallow processing; and, for remembered words only, for low- than for high-frequency words. Latency and topographic differences between these potentials suggested that several factors influence the effectiveness of encoding and can be distinguished using these methods, even with Chinese logographic symbols. PMID- 14746869 TI - Differential processing of duration changes within short and long sounds in humans. AB - It has been postulated that there exist two different mechanisms of duration processing. Brief durations, shorter than 500 ms, are processed sensorially, whereas longer durations are processed via a cognitive mechanism. We investigated whether electrophysiological evidence for this distinction can be found. In particular, we measured the mismatch negativity (MMN) to auditory duration deviants (40% decrements) in blocks of short (200 ms) and long sounds (1000 ms) in Ignore and in Attend conditions. Our results show a typical MMN for long and short durations in the Attend condition, whereas no MMN was obtained for long durations in the Ignore condition. This interaction between duration and attention seems to reflect a breakdown of the sensorial mechanism of temporal processing at some critical duration of about 1 s when sounds are ignored. PMID- 14746870 TI - Repetitive transcranial magnetic stimulation of the motor cortex attenuates pain perception in complex regional pain syndrome type I. AB - In complex regional pain syndrome (CRPS) many clinical symptoms suggest involvement of the central nervous system. Neuropathic pain as the leading symptom is often resistant to therapy. In the present study we investigated the analgesic efficiency of repetitive transcranial magnetic simulation (rTMS) applied to the motor cortex contralateral to the CRPS-affected side. Seven out of ten patients reported decreased pain intensities. Pain relief occurred 30 s after stimulation, whereas the maximum effect was found 15 min later. Pain re intensified increasingly 45 min after rTMS. In contrast, sham rTMS did not alter pain perception. These findings provide evidence that in CRPS I pain perception can be modulated by repetitive motor cortex stimulation. PMID- 14746871 TI - Sustained increase of somatosensory cortex excitability by 5 Hz repetitive transcranial magnetic stimulation studied by paired median nerve stimulation in humans. AB - Repetitive transcranial magnetic stimulation (rTMS) has been shown to alter cortical processing within primary motor cortex dependent on the choice of stimulation variables. However, little is known about the effects of TMS in other cortical areas such as the primary somatosensory cortex (SI). Here we asked whether high-frequency (5 Hz) rTMS applied over the left SI evokes sustained changes in cortical excitability. To assess excitability changes, we applied a paired-pulse protocol consisting of paired electrical stimulation of the median nerve using an interstimulus interval of 30 ms and recordings of somatosensory evoked potentials. For ipsilateral SI we found that 1 h after termination of 5 Hz rTMS applied over the left SI with a figure-of-eight coil there was a sustained suppression of the normally present paired-pulse inhibition. Latencies and N20 amplitudes of the first peak remained unchanged. No changes of paired-pulse behavior were observed in the contralateral SI that was not TMS stimulated. The sustained excitability enhancement in SI is discussed in respect to previous findings regarding an improvement of tactile discrimination behavior by rTMS. PMID- 14746872 TI - Prenatal and neonatal exposure to bisphenol-A affects the morphine-induced rewarding effect and hyperlocomotion in mice. AB - Bisphenol-A (BPA), one of the most common environmental endocrine disrupters, has been extensively evaluated for toxicity and carcinogenicity. However, little is still known about its action on the CNS. Here we found that prenatal and neonatal exposure to BPA resulted in the enhancement of the rewarding effect and hyperlocomotion induced by morphine in mice. Under these conditions, no change in the G-protein activation by morphine and mu-opioid receptor expression in the lower midbrain was observed by prenatal and neonatal exposure to BPA. These results suggest that chronic exposure to BPA produces the supersensitivity of the morphine-induced rewarding effect and hyperlocomotion without direct changes in mu-opioid receptor function in the lower midbrain. The present data provide further evidence that prenatal and neonatal exposure to BPA can directly influence the development of the central dopaminergic system. PMID- 14746873 TI - Effects of electroacupuncture on gastric myoelectric activity and substance P in the dorsal vagal complex of rats. AB - Acupuncture has been widely applied to treat gastrointestinal diseases in the clinic. However, its precise mechanism remains unknown. In the present study, effects of electroacupuncture (EA) at Zusanli acupoints (ST36) on gastric myoelectric activity and substance P (SP) in the dorsal vagal complex (DVC) were observed. EA at ST36 significantly increased the frequency of slow waves and the frequency of the spike potentials superimposed on the slow waves, which persisted for 90 min after cessation of EA and was completely abolished after bilateral vagotomy. Meanwhile, the mean optical density of SP immunoreactive fibers in the DVC increased following EA. The results suggested that EA at ST36 promoted the gastric myoelectric activity, which was regulated by the vagus, and SP in the DVC may be involved in the excitatory effects. Furthermore, EA had a relative long term post-effect. We presumed that DVC may be a supraspinal center in the effect of EA on gastric function. PMID- 14746874 TI - Enhanced re-habituation of the orienting response of the human event-related potential. AB - Previous studies found the amplitude of the orienting response (OR) of the human event-related potential to decrease with repeated stimulus presentations. This decrease has been suggested to reflect short-term habituation and/or long-term habituation, both of which are learning processes. However, this earlier research failed to provide direct evidence supporting this claim. The present study attempted to show that the OR pattern shares one important feature of habituation: an enhanced response decrement across stimulus-presentation blocks (enhanced re-habituation). Participants received four blocks of 25 auditory stimulus presentations and showed an OR decrement both within (short-term habituation) and across (long-term habituation) blocks. Importantly, the OR decreased more rapidly during later than initial trial blocks, suggesting enhanced re-habituation. The latter result supports the notion that the amplitude decrement reflects an elementary learning process. PMID- 14746875 TI - Identification of the mouse neurochondrin promoter region and the responsible region for cell type specific gene regulation. AB - Neurochondrin is a cytoplasmic protein possibly involved in neurite outgrowth and chondrocyte differentiation. In the present study, we have identified 202 bp of the mouse neurochondrin minimal promoter sequences encompassing the transcriptional initiation site, and both of the activating and repressing regions in the first exon. These two regulatory regions in the first exon had a cell type dependent effect on the identified minimal promoter. In the regulatory region, the duplication of potential binding sites for GATA family transcriptional factors was observed. Prospective binding sites for sex determining region Y and c-Ets1 were also found in the minimal promoter region. These factors could be potential regulators for the mouse neurochondrin gene. PMID- 14746876 TI - Microglia-derived glial cell line-derived neurotrophic factor could protect Sprague-Dawley rat astrocyte from in vitro ischemia-induced damage. AB - The interrelationship between microglia and astrocytes in cerebral ischemia was determined in vitro by adding in vitro ischemia-induced supernatant from microglia into astrocytes under the same conditions (glucose-, oxygen- and serum free). The involvement of glial cell line-derived neurotrophic factor (GDNF) was further investigated by immunoblocking assay and Western blot analysis. Results showed that microglia-derived supernatant protected against in vitro ischemia induced damage of astrocytes and this protection was pre-blocked by anti-GDNF but not normal rabbit serum. In addition, in vitro ischemia appeared to induce the expression of GDNF in microglia. These results indicate that microglia-derived protection on astrocytes during in vitro ischemia is GDNF-dependent. PMID- 14746877 TI - Early versus late bedtimes phase shift the human dim light melatonin rhythm despite a fixed morning lights on time. AB - Short sleep/dark durations are common in modern society. In a within subjects counterbalanced design, we compared the effect of 7-19 nights with a late bedtime (01:00) versus 7-19 nights with an early bedtime (22:00) on the human dim light melatonin rhythm. All ten healthy subjects experienced lights on time at 07:00 and at least 5 min of outdoor light between 07:00 and 08:00. We found that despite the morning light exposure, the melatonin rhythm following the late bedtime nights was delayed by approximately 0.6 h (P<0.05), as compared to after the early bedtime nights. These results suggest that when people truncate their sleep by delaying their bedtime they also significantly phase delay their circadian rhythms and/or when people advance their bedtime their circadian rhythms advance. PMID- 14746878 TI - Promoter haplotypes of interleukin-10 gene and sporadic Alzheimer's disease. AB - Clinical and immunopathological evidence support a potential role of the pro- and anti-inflammatory cytokine network in neurodegeneration of Alzheimer's disease (AD). Moreover, association studies suggest a possible involvement of cytokine related genes in the susceptibility to sporadic AD. Since conflicting results are associated with the pro-inflammatory pathway, we investigated a putative effect of the anti-inflammatory counterpart focusing on the interleukin-10 (IL-10) gene. The 5' flanking region contains numerous polymorphisms; in particular, three single nucleotide polymorphisms (-1082 G/A, -819 T/C, -592 C/A) are in linkage disequilibrium resulting in three haplotypes GCC, ACC and ATA. We analyzed the IL 10 haplotype distributions in 215 Italian sporadic AD patients and 153 controls in an association case-control study. Haplotype frequencies did not reveal differences between the two samples, however the genotype GCC/ACC was more represented in AD patients (OR 1.91, 95% CI: 1.18-3.07). This putative risk factor could be independent of the presence of the ApoE epsilon 4 allele. Our results provide new insights on a possible involvement of the IL-10 gene in susceptibility to sporadic AD even though further functional and genetic investigations are necessary to clarify its role in AD. PMID- 14746879 TI - Sodium salicylate-induced amelioration of experimental autoimmune encephalomyelitis in Lewis rats is associated with the suppression of inducible nitric oxide synthase and cyclooxygenases. AB - We studied the effects of oral administration of sodium salicylate on the expression of the pro-inflammatory mediators, nitric oxide synthase (iNOS) and cyclooxygenase- (COX-) 1 and 2, in rats with experimental autoimmune encephalomyelitis (EAE). Sodium salicylate (200 mg/kg) was administered orally for 13 days after the induction of EAE by immunization with guinea pig myelin basic protein and complete Freund's adjuvant. The onset (P<0.0001) and severity (P<0.05) of EAE paralysis in salicylate-treated animals were delayed and suppressed significantly compared with vehicle-treated controls. Western blot analysis showed that expression of COX-2 and iNOS, but not COX-1, decreased significantly in the spinal cords of salicylate-treated rats compared with vehicle-treated controls (P<0.05) and this finding was paralleled by immunohistochemical observations. These results suggest that the amelioration by salicylate of paralysis in rats with EAE is mediated in part by the suppression of COX and iNOS. PMID- 14746880 TI - Dopamine modulation of glutamate metabotropic receptors in conditioned reaction of sensory motor cortex neurons of the cat. AB - The dopamine modulation of microiontophoretic application of antagonists of glutamate metabotropic synaptic transmission was studied in the sensory motor cortex of awake cats during instrumental conditioned reflex. The substances depressed the background and intensity of evoked impulse activity of pyramidal neurons of the sensory-motor cortex and provoked significant increases in the latency of impulse reaction and corresponding conditioned movements of animals. Simultaneous application of the antagonists and dopamine (DA) eliminated their depression of the background and evoked activity of neurons and decreased the latency of the impulse reactions and movements. Similar qualitative effects were observed in experiments with simultaneous application of biccuculine and (RS) alpha-methyl-4-carboxyphenylglycine. It is supposed that the DA modulation in the brain consists in stabilizing the background and evoked activity of cortical neurons during reduced intensity of metabotropic glutamatergic synaptic transmission. Such modulation can be important when considering some pathological disorders of glutamatergic transmission. PMID- 14746881 TI - Increased dopamine transporter density in the male rat brain following chronic nandrolone decanoate administration. AB - Adolescent males currently employ anabolic-androgenic steroids (AAS) to become intoxicated, besides the traditional desires of an improved physical appearance and enhanced sports performance. Several studies indicate that AAS affect the brain reward system. Recently chronic administration with nandrolone decanoate to male rats has been shown to increase the dopamine transporter (DAT) density in the striatum visualised in vivo by positron emission tomography. The present study aimed to investigate if the increased DAT density could be confirmed using in vitro autoradiography following a comparable regimen of nandrolone treatment. Specific binding of 50 pM [125I] RTI-55 in the presence of 1 microM citalopram was used to label DAT. Two weeks of nandrolone decanoate administration at the supra-therapeutic doses 1, 5 and 15 mg/kg per day increased DAT density in the caudate putamen at all three doses. In conclusion, this study confirms that chronic nandrolone administration increases dopamine transporter density in the CPU and therefore supports the theory that AAS affects the dopamine system in the male rat brain. PMID- 14746882 TI - Olanzapine counteracts reduction of brain-derived neurotrophic factor and TrkB receptors in rat hippocampus produced by haloperidol. AB - Recently, we have reported studies in rats showing that the atypical antipsychotic olanzapine (OLZ), in contrast to haloperidol (HAL), was not associated with reduction of markers of central cholinergic neurons as well as decrements in cognitive performance after chronic exposure. We compared the effect of 45 day chronic exposure of HAL (2 mg/kg per day) to OLZ (10 mg/kg per day) on brain-derived neurotrophic factor (BDNF)) and its high affinity receptor TrkB in rat hippocampus. Since the use of OLZ is presently preferred over HAL in patients, effects of its post-treatment on HAL-induced changes in the expression of BDNF and its TrkB receptor were also investigated. OLZ was administered after 45 days of HAL exposure. HAL, but not OLZ, significantly reduced the levels of BDNF in hippocampus. These changes in BDNF paralleled the levels of TrkB receptors. Furthermore, post-treatment with OLZ markedly restored the HAL treatment associated reductions in both BDNF and TrkB receptors in hippocampus. PMID- 14746883 TI - Optic nerve ligation leads to astrocyte-associated matrix metalloproteinase-9 induction in the mouse retina. AB - Ischemic damage results in irreversible ganglion cell loss in the retina. While the mechanisms underlying ischemia-induced ganglion cell loss are not clearly understood, we have recently reported that ischemia, induced by optic nerve ligation, results in increased nerve fiber layer-associated matrix metalloproteinase-9 (MMP-9) induction and loss of ganglion cells from the retina. This study was conducted to determine the cellular source of MMP-9 using antibodies against MMP-9 and various cell types in the inner retina. The results presented in this study show that optic nerve ligation leads to induction of MMP 9 and activation of astrocytes. Double labeling studies using antibodies against MMP-9 and GFAP showed a greater overlap of MMP-9 with GFAP, compared to antibodies against glutamine synthetase and MMP-9 which showed no overlapping, suggesting that activated astrocytes contribute to MMP-9 expression in the retina. Further, double labeling studies using antibodies against von Willebrand factor and MMP-9 or Mac-1 and MMP-9 showed no overlapping of MMP-9 with any antibodies indicating that endothelial cells and microglial cells are not the principal source of MMP-9 in the retina following optic nerve ligation. PMID- 14746884 TI - Region-dependent G-protein activation by kappa-opioid receptor agonists in the mouse brain. AB - The G-protein activations induced by kappa-opioid receptor agonists, (-)U50,488H, U69,593 and TRK-820 in the mouse lower midbrain, striatum and limbic forebrain were determined by monitoring guanosine-5'-o-(3-[35S]thio)triphosphate ([35S]GTP gamma S) binding. All kappa-opioid receptor agonists produced approximately 40, 20 and 10% increases of [35S]GTP gamma S binding over baseline in the lower midbrain, striatum and limbic forebrain, respectively. The increases of [35S]GTP gamma S binding induced by kappa-opioid receptor agonists were completely reversed by the selective kappa-opioid receptor antagonist, norbinaltorphimine (norBNI), in all brain regions. The intrinsic activities of kappa-opioid receptor agonists for G-protein activation in brain regions observed in the present study are not correlated with densities of kappa-opioid receptor binding sites from previous reports. The present results suggest that the catalytic efficiency of kappa-opioid receptor-G-protein coupling may be variable in different brain regions. PMID- 14746885 TI - Hypoxia stimulates glutamate uptake in whole rat retinal cells in vitro. AB - Glutamate uptake in neurons and glial cells is essential to prevent the persistence of excitotoxic levels of glutamate observed during ischemia. We demonstrated that a short period of hypoxia stimulated the apparent glutamate transport rate in isolated rat retinal cells. The observed increase in glutamate uptake was not affected by glutamate receptor antagonists, protein kinase inhibitors, antioxidant or neo-synthesis inhibitors. However, inhibition of actin polymerization reversed the hypoxia-induced increase in glutamate uptake, suggesting a mobilization of transporters to the cell membrane. Moreover, the depletion in cell glutathione stimulated in the same manner the glutamate uptake and emphasized the key role of glutamate in the control of the level of this antioxidant. This rapid up-regulation of glutamate transport could be considered as an adaptative mechanism of neuroprotection. PMID- 14746886 TI - Wide-range visualization of compound nerve action magnetic fields in the human median and ulnar nerves from the forearm to Erb's point. AB - We successfully visualized the compound nerve action magnetic fields (CAFs) in the human median and ulnar nerves from the forearm to Erb's point. To observe the CAFs, we used a superconducting quantum interference device gradiometer system that was developed for human peripheral nerves. The CAFs were visualized as a quadrupole pattern consisting of leading and trailing dipoles. The CAFs propagated along the anatomical pathway and extended longitudinally in the proximal segment. The most reasonable explanation is that the peak separation in the trailing dipole appeared when the leading dipole reached the proximal segment after stimulation of the median nerve at the wrist. A temporal dispersion of the CAFs was suggested to be visualized. PMID- 14746887 TI - Modulation of cognitive processing by emotional valence studied through event related potentials in humans. AB - This experiment investigated whether the emotional content of a stimulus could modulate its cognitive processing. Particularly, we focused on the influence of the valence dimension on the cognitive processing triggered by a non emotional oddball task. To this end, event-related potentials (ERPs) were recorded from 25 sites during a visual oddball paradigm. Three sets of pictures (unpleasant, neutral and pleasant) with low arousal values served as rare target items. Subjects were simply asked to realize a standard/target categorization task, irrespective of the picture valence. A temporal principal component analysis allowed us to identify several evoked components (i.e. P1, P2, N2, P3a and P3b). Emotional effects observed on P1, P2 and P3b showed that the valence content of the stimulus modulates the cognitive processes at several points in the information processing stream. PMID- 14746888 TI - Potentiation of mitogenesis in adult rat chromaffin cell cultures by immunosuppressive agent FK506. AB - The immunosuppressive drugs FK506 and cyclosporin inhibit T- and B-lymphocyte proliferation and exert neuritogenic and/or cytoprotective effects on several types of neurons. While the immunosuppressive actions of both drugs are mediated in large part by inhibition of the Ca(2+)-dependent phosphatase, calcineurin, FK506 is known to exert additional effects. In the present study, FK506 is shown to potentiate mitogenic effects of the neurotrophic factor, neurturin, on normal adult rat adrenal chromaffin cells in culture. The effect is not seen with cyclosporin or with a non-immunosuppressive analog of FK506, GPI-1046. The finding of increased mitogenesis in response to FK506 may have applications to the study of normal and neoplastic neuroendocrine cells and to understanding the development of some types of tumors in transplant patients. PMID- 14746889 TI - Inhibitory effect of intrathecal meptazinol on carrageenan-induced thermal hyperalgesia in rats. AB - The effect of meptazinol in the spinal cord on carrageenan-induced hyperalgesia was investigated. The latency of paw withdrawal (PWL) to a thermal stimulus was used as an index of inflammatory hyperalgesia in awake rats. Intrathecal (i.t.) injection of 10 and 100 microg meptazinol markedly increased the PWL of the carrageenan-injected paw (P<0.01). The PWL of the non-injected paw was not detectably affected by the administration of meptazinol at the doses tested. I.t. injection of naloxone (5 microg) or atropine (1 microg) alone exhibited no effect on the PWLs of either the carrageenan-injected or non-injected paw. Pretreatment with naloxone, but not atropine, completely blocked the meptazinol-induced anti hyperalgesia. These observations suggested that mu opioid receptor rather than muscarinic acetylcholine receptor may be involved in the anti-hyperalgesia of meptazinol in the spinal cord. PMID- 14746890 TI - The somatostatin receptor (sst1) modulates the release of somatostatin in rat retina. AB - The aim of this study was to examine the ability of somatostatin receptor (sst(1)) to regulate the release of somatostatin in rat retina. Immunohistochemistry studies were performed to locate the somatostatin neurons, and radioligand binding to ascertain the presence of sst(1). The neuronal release of somatostatin was examined ex vivo in rat retinal explants in the presence of KCl (50 and 100 mM), and absence of Ca(++) (EGTA; 10 mM). Somatostatin levels, quantified by radioimmunoassay, were increased in the presence of KCl (100 mM, 151%) and attenuated in the absence of Ca(++) (31%). CH275 (sst(1) agonist) reduced the somatostatin levels in a concentration-dependent manner (10(-5)-10( 7) M), and this effect was reversed by NVP-SRA 880 (sst(1) antagonist;10(-5) M). MK678 (sst(2) agonist; 10(-5) M) had no effect. These data suggest an autoreceptor role for sst(1) in retina. PMID- 14746891 TI - Reticulon3 expression in rat optic and olfactory systems. AB - Reticulon3 (RTN3), which belongs to a reticulon family, is first isolated from the retina, but little is known about its function. We investigated the distribution of RTN3 in rat retina and olfactory bulb by immunohistochemistry. In the retina, Muller cells highly expressed RTN3. The expression level of RTN3 in the optic nerve was high in the embryo, but low in the adult. In the olfactory system, RTN3 was highly expressed in the olfactory nerve both in developmental and adult stages. Further, RTN3 was co-localized with synaptophysin in tubulovesicular structures in the developing axon of cultured cortical neurons. These results suggest that RTN3 may play an important role in the developing axons and also in some glial cells such as Muller cells. PMID- 14746892 TI - Inhibition of dopamine synthesis with alpha-methyl-p-tyrosine abolishes the enhancement of methamphetamine-induced extracellular dopamine levels in the amygdala of rats with excitotoxic lesions of the entorhinal cortex. AB - This study was performed to investigate the mechanisms underlying the augmentation of methamphetamine (MAP)-induced dopamine (DA) release in the entorhinal cortex-lesioned rats. Quinolinic acid or phosphate buffered saline was infused into the left entorhinal cortex of adolescent rats (postnatal day 7 weeks). After 4 weeks of lesioning, acute MAP (2 mg/kg, i.p.)-induced DA release in the amygdala was significantly enhanced in lesioned rats compared to sham operated rats. Inhibition of DA synthesis by alpha-methyl-p-tyrosine, an inhibitor of catecholamine synthesis, resulted in abolishment of the enhancement of MAP (2 or 5 mg/kg, i.p.)-induced DA release in the amygdala of lesioned rats. These results suggest that excessive DA pool in nerve terminals underlies the augmentation of MAP-induced DA release in the amygdala of the lesioned rats. PMID- 14746893 TI - Hydrogen peroxide modulates whole cell Ca2+ currents through L-type channels in cultured rat dentate granule cells. AB - Modification of voltage-gated Ca(2+) channels by hydrogen peroxide, a membrane permeable form of reactive oxygen species, in cultured dentate granule cells was examined using the whole cell patch clamp technique. Pretreatment with hydrogen peroxide (1 and 10 microM) for 2 h enhanced the Ca(2+) current without affecting its voltage dependence. The enhancement was completely cancelled by 1 mM glutathione, an antioxidant, and 2 microM nifedipine, an L-type Ca(2+) channel blocker. In contrast, the enhancement of the Ca(2+) current was not mimicked by pretreatment with 10 microg/ml tunicamycin, an endoplasmic reticulum stressor. These results suggest that oxidative stress induced by hydrogen peroxide selectively regulates the activity of L-type Ca(2+) channels. PMID- 14746894 TI - Postsynaptic metabotropic glutamate receptor mGluR1 mediates the late component of signal propagation in the guinea pig piriform cortex: optical imaging study. AB - Metabotropic glutamate receptors (mGluRs) were previously shown to mediate a postsynaptic late propagation component elicited by layer Ib stimulation in guinea pig piriform cortex slices. In the present study, the effects of some group specific or subtype specific mGluR antagonists on the late propagation component were investigated using an optical imaging method, in order to identify mGluR subtypes mediating it. A selective mGluR1 antagonist (RS)-1-aminoindan-1,5 dicarboxylic acid most effectively suppressed the late component whereas a selective mGluR5 antagonist, selective group II or group III antagonists showed little or no suppressive effect. These results suggest that the late propagation component is mediated by mGluR1. PMID- 14746895 TI - The alpha 1D-adrenergic receptor modulates cardiovascular and drinking responses to central salt loading in mice. AB - To characterize the involvement of specific alpha(1)-adrenergic receptor (alpha(1)-AR) subtypes in hypertension, parameters related to central salt- or angiotensin II (ANG II)-induced hypertension were investigated in alpha(1D)-AR deficient mice (knockout). Baseline daily water intake and food intake were larger in alpha(1D)(-/-) mice than in alpha(1D)(+/+) mice. Intracerebroventricular (i.c.v.) administration of NaCl (0.67 M NaCl, 1 microl) elicited smaller increases in mean arterial blood pressure (MABP), heart rate, and water intake in alpha(1D)(-/-) mice than it did in alpha(1D)(+/+) mice. I.c.v. administration of ANG II (10 pmol) resulted in increases in MABP and water intake that were similar in alpha(1D)(-/-) mice and alpha(1D)(+/+) mice. These results suggest that alpha(1D)-AR is, at least in part, involved in central salt induced but not ANG II-induced hypertension and water intake. PMID- 14746896 TI - Peri-sciatic administration of indomethacin early after nerve injury can attenuate the development of tactile allodynia in a rat model of L5 single spinal nerve injury. AB - To clarify the role of cyclooxygenase in the peripheral nerve on the development of neuropathic pain, we investigated the effects of peri-sciatic administration of indomethacin on the development of allodynia in a model of L5 single spinal nerve injury. Peri-sciatic administration of indomethacin (1 mg/kg) was performed 3, 24, or 72 h after nerve injury (n=6/each). In rats with indomethacin 3 or 24 h after nerve injury, ipsi-lateral paw withdrawal thresholds 7-35 days after nerve injury were significantly higher compared with those in the control group (n=6: without peri-sciatic treatment) (P<0.05). However, such efficacy was no longer apparent when indomethacin was administered 72 h after nerve injury. These results suggest that peri-sciatic administration of indomethacin early (less than 24 h) after nerve injury can attenuate the development of allodynia. PMID- 14746897 TI - Pilocarpine-induced status epilepticus increases glutamate release in rat hippocampal synaptosomes. AB - A pronounced glutamate release has been related to neuronal death in several structures due to status epilepticus (SE). We investigated the glutamate uptake and release by both cortical and hippocampal synaptosome in pilocarpine model of epilepsy. Animals were submitted to long-lasting SE (12 h) induced by pilocarpine and compared with non-treated animals. Animals presenting SE did not modify the glutamate uptake by synaptosomes. An increase in the glutamate efflux in the absence (1.43-fold) and in the presence of KCl (1.25-fold) was found in hippocampal synaptosomes. Pilocarpine added to the medium did not modify the glutamate release profile, showing that SE is necessary to modify the glutamate release. As the glutamate uptake is not modified, the hippocampal excitotoxicity may be related to impairment only in the mechanism of the glutamate release. PMID- 14746898 TI - Effect of poly(ADP-ribose) polymerase inhibitors on oxidative stress evoked hydroxyl radical level and macromolecules oxidation in cell free system of rat brain cortex. AB - Poly(ADP-ribose) polymerase-1 (PARP-1) is a nuclear enzyme involved in DNA repair, replication and cell cycle. However, its overactivation leads to nicotinamide adenine dinucleotide and ATP depletion and cell death. The inhibitors of PARP-1 were successfully used in the basic studies and in animal models of different diseases. For this reason, it is important to discriminate between specific and non-specific antioxidant properties of PARP-1 inhibitors. The aim of this study was to investigate the effect of PARP-1 inhibitors on the free radical level and oxidation of macromolecules and to compare their properties with the efficacy of antioxidants. Oxidative stress was induced in the brain cortex homogenate by FeCl(2) or CuSO(4) at 25 microM during 15 min incubation at 37 degrees C. PARP-1 inhibitors 3-aminobenzamide (3-AB), 1,5 dihydroxyisoquinoline (DHIQ) and 3,4-dihydro-5-[4-(1-piperidinyl)butoxy]-1(2H) isoquinolinone (DPQ), and the antioxidants alpha-tocopherol, resveratrol and Tempol were used at 0-5 mM. Free radical contents were estimated by spin-trapping using HPLC. Lipid and protein oxidation were determined by measuring thiobarbituric acid reactive substances and carbonyl groups or using fluorescent probe TyrFluo, respectively. Our data indicate that 3-AB and DHIQ are potent hydroxyl radical scavengers and inhibitors of protein oxidation. DHIQ additionally decreases lipid peroxidation. DPQ has no antioxidant properties and seems to be a specific PARP-1 inhibitor, however, it is a water insoluble compound. Among the investigated antioxidants, the most potent was resveratrol and then alpha-tocopherol and Tempol. These results indicate that 3-A beta, benzamide and DHIQ are potent hydroxyl radical scavengers and antioxidants. These data ought to be taken into consideration when properties of these compounds as PARP inhibitors are evaluated. PMID- 14746900 TI - Spatiotemporal changes of the N-methyl-D-aspartate receptor subunit levels in rats with pentylenetetrazole-induced seizures. AB - The aim of this study was to examine the expression profiles of N-methyl-D aspartate (NMDA) receptor subunits in rats during seizure development and kindled process induced by pentylenetetrazole (PTZ). Using quantitative Western blotting, the levels of NR1, NR2A and NR2B subunits were measured in the cortex and hippocampus of rats at different times after PTZ injection. In the early seizure developmental process, both NR1 and NR2A were markedly increased in the cortex, and NR1 was significantly increased in the hippocampus. On the other hand, in the kindled process both NR1 and NR2A decreased in the cortex and hippocampus. However, the NR2B subunit had no appreciable change in both the seizure developmental and kindled process. Therefore, these results showed that expression of NMDA receptors undergoes subunit- and region-related changes in the developmental and kindled seizure of rats induced by PTZ. PMID- 14746899 TI - A human granin-like neuroendocrine peptide precursor (proSAAS) immunoreactivity in tau inclusions of Alzheimer's disease and parkinsonism-dementia complex on Guam. AB - The deposition of tau inclusions is one of the neuropathological hallmarks in neurodegenerative disorders with dementia. We have reported that the N-terminal fragment of a human granin-like neuroendocrine peptide precursor (N-proSAAS) is accumulated in Pick bodies. However, it is unknown whether N-proSAAS is widely accumulated in tau inclusions in other tauopathies. Here, we performed an immunohistochemical examination using antibodies against both the N- and C terminal sequence of proSAAS in the brains of patients with Alzheimer's disease and parkinsonism-dementia complex on Guam. The antibody against N-proSAAS immunostained neurofibrillary tangles and neuritic plaques in both diseases, whereas the antibody against the C-terminal sequence of proSAAS did not. The results of the present study suggest that N-proSAAS or proSAAS-like molecules were trapped within the tau fibrils and accumulated in tau inclusions. PMID- 14746901 TI - Picrotoxin increased acetylcholine release from rat cultured embryonic septal neurons. AB - GABA is a major inhibitory neurotransmitter in the mature mammalian brain. In the early stages of brain development, it has been reported that GABA(A) receptor stimulation and the associated increase in Cl(-) conductance lead to membrane depolarization. In this study, we tested the effects of picrotoxin, a GABA(A) receptor Cl(-) channel blocker, on spontaneously released acetylcholine (ACh) from cultured rat embryonic septal cells. Picrotoxin increased spontaneously released ACh. These results indicate that blockade of GABA-activated Cl(-) channel increases neuronal excitability even in an early stage of the development. PMID- 14746902 TI - Do bimanual isometric push efforts in humans stop as a consequence of postural muscle exhaustion? AB - The purpose of this study was to explore whether global efforts stop as a consequence of postural muscle exhaustion. To this end, seated adults were asked to exert 75% maximal voluntary contractions bimanual push efforts until exhaustion. A dynamometer was used to measure the horizontal force exerted on a bar (Fx) and a custom-designed force plate measured the antero-posterior displacement of the centre of pressure (Xp). Electromyograms were picked up by bipolar surface electrodes from the primum movens (serratus anterior) and four postural muscles (trapezius superior, erectores spinae, rectus abdominis, rectus femoris). Root mean square and mean power frequency were calculated over 2-s intervals and compared to corresponding Fx and Xp values. It was shown that the effort stops as a consequence of exhaustion of postural muscles (rectus abdominis and rectus femoris), and not of the primum movens. It is concluded that postural muscles make a major contribution to global efforts, in that they allow compliance to biomechanical requirements, that is, to preserve the distance between the centre of pressure and the centre of gravity, which must be proportional to the external force. PMID- 14746903 TI - Diazepam prophylaxis for bicuculline-induced seizures: a rat dose-response model. AB - We developed a screening methodology to test the ability of putative antiepileptic drugs delivered directly to a seizure focus to prevent epileptiform activity. The left hippocampi of 15 rats were implanted with an injection cannula and bipolar recording electrodes. Bone screws were used to record neocortical EEG activity. Diazepam (DZP) at one of four possible concentrations or control solution was injected into the hippocampus, followed 5 min later by bicuculline methiodide. DZP suppressed spikes and ictal events in a dose-dependent manner (P<0.0001). At 100 mM, DZP reduced spikes from 678+/-128 to 87+/-35 for a 15 min segment. Numbers of ictal events (seizure) and latency to the first event were reduced by prophylactic DZP. The study establishes a protocol for testing of intracranially-injected drugs to prevent focal seizures. PMID- 14746904 TI - Effect of zinc in ischemic brain injury in an embolic model of stroke in rats. AB - Zinc is prevalent in the mammalian central nervous system and its role in ischemic brain injury is still controversial. In the present study, the effect of zinc in ischemic brain injury was examined in an embolic model of stroke in rats. Furthermore, the effect of zinc in combination with bicuculline, a GABAa antagonist, was also examined in the ischemic injury. Treatment with zinc or zinc plus bicuculline increased infarct volume significantly and also worsened neurological deficits. Moreover, treatment with zinc plus bicuculline also enhanced ischemic brain edema. These results thus support the hypothesis that administration of zinc i.p. worsens the outcome of ischemic brain injury in the embolic model of stroke in rats. PMID- 14746905 TI - Immunohistochemical detection of osteopontin in the spinal cords of mice with Theiler's murine encephalomyelitis virus-induced demyelinating disease. AB - The spinal cords of mice that were infected with the BeAn 8386 strain of Theiler's murine encephalomyelitis virus (TMEV) were studied to elucidate the involvement of osteopontin in the course of TMEV-induced demyelination. Immunohistochemistry showed staining for osteopontin in the vessels of the normal spinal cords, and more intense immunoreactivity in the vessels within the demyelinating lesions. Intense osteopontin immunoreactivity was observed in the cell bodies, as well as in the extracellular space of the demyelinating lesions, where some glial cells, which included activated microglia/macrophages, were also immunopositive for osteopontin. These findings suggest that osteopontin is upregulated in the demyelinating spinal cord, and that osteopontin from either microglia or astrocytes may be involved in the chemotaxis of inflammatory cells and astrocytes, which ultimately leads to chronic inflammation and astrogliosis in this model system. PMID- 14746906 TI - Identification and characterization of mouse TRMU gene encoding the mitochondrial 5-methylaminomethyl-2-thiouridylate-methyltransferase. AB - The nucleotide modification in tRNA plays a pivotal role in the fidelity of translational process. The mutated mitochondrial tRNA (mt tRNA) associated with human diseases often exhibited a defect in nucleotide modification at wobble position of anticodons. Recently, the product of trmU, 5-methylaminomethyl-2 thiouridylate-methyltransferase, has been shown to be one component of enzyme complex for the biosynthesis of mnm5s2U in the wobble position of the bacterial tRNAs. Here we report the identification and characterization of mouse TRMU homolog. A 1532 bp TRMU cDNA has been isolated and the genomic organization of TRMU has been elucidated. The mouse TRMU gene containing 11 exons encodes a 417 residue protein with a strong homology to the TRMU-like proteins of bacteria and other homologs related to tRNA modification. The mouse TRMU is ubiquitously expressed in various tissues, but abundantly in tissues with high metabolic rates including heart, liver and brain. Furthermore, immunofluorescence analysis of NIH3T3 cells expressing TRMU-GFP fusion protein demonstrated that the mouse Trmu localizes in mitochondria. These observations suggest that the mouse TRMU is a structural and functional homolog of bacterial TrmU, thereby playing a role in the mt tRNA modification and protein synthesis. PMID- 14746907 TI - Mouse keratinocytes express c98, a novel gene homologous to bcl-2, that is stimulated by insulin-like growth factor 1 and prevents dexamethasone-induced apoptosis. AB - Many studies have been undertaken to investigate the mechanisms of skin differentiation. In particular, growth factors and hormones are believed to play important roles in skin proliferation, differentiation and survival. Insulin-like growth factor-1 (IGF-1) has been identified as a survival factor in many tissues including the skin, but the molecular mechanism of IGF-1 in epidermal differentiation is not completely understood. Neonatal mouse skin is useful for studying changes in gene expression, as the mitotic activity of skin cells changes shortly after birth. Using RNA differential display (DD), a 357-nt message that is specifically expressed in the epidermal keratinocytes of IGF-1 injected newborn mice but not in controls, has been identified. Confirmation of expression of this gene by ribonuclease protection assay (RPA) showed that its mRNA expression in the epidermal keratinocytes is induced by IGF-1. Using RNA ligase-mediated rapid amplification of 5' cDNA ends (RLM-5'-RACE), we have successfully isolated a 3473-bp full-length gene, c98, that has 97% sequence homology to a bcl-2-like gene, bcl-w. The latter has been identified as a proto oncogene in several murine myeloid cell lines. Amino acid sequence analysis of the c98 showed that it has 97% sequence identity to the bcl-w protein and possesses bcl-2 homology domains (BH) 1, 2 and 3. Immunoblotting data revealed similar increases of c98 protein expression to its mRNA expression in the keratinocytes of IGF-1-injected animals. Weak expression of other bcl-2 family member proteins, bax, bcl-2 and bcl-xL, were also found in the immunoblots. Additionally, IGF-1 was found to be able to protect epidermal keratinocytes from dexamethasone (DEX)-induced apoptosis, based on the findings that after the cells were treated with DEX, DNA laddering was present in the control mice but not in those injected with IGF-1. Further, using a photometric enzyme-linked immunoassay to quantitate keratinocyte death, we found that after addition of DEX, the amounts of cytoplasmic histone-associated DNA fragments were not significantly (P>0.05) different in IGF-1-treated cells compared with untreated control cells during the high mitotic stage of skin epidermis. To assess the role of c98 in these anti-apoptotic processes, we have generated a recombinant plasmid that contains an expression vector and c98 and transfected this plasmid into the keratinocytes from mice without IGF-1-treatment. Expression of the c98 protein was found to completely (P>0.05) block DEX-induced apoptosis after cell transfection. Taken together, our current data demonstrated that IGF-1 plays an anti-apoptotic role in the DEX-induced apoptosis in epidermal keratinocytes and this, at least in part, may be mediated through expression of c98. PMID- 14746908 TI - Intramuscular plasmid DNA electrotransfer: biodistribution and degradation. AB - We have studied radiolabelled plasmid DNA biodistribution and degradation in the muscle at different times after injection, with or without electrotransfer using previously defined conditions. Radiolabelled plasmid progressively left the muscle and was degraded as soon as 5 min after plasmid injection, with or without electrotransfer. Autoradiography showed that the major part of injected radioactivity was detected in the interfibrilar space of a large proportion of the muscle. Large zones of accumulation of radioactivity, which seems to be contained in some fibres (more than 20 microm), were identified as soon as 5 min after electrotransfer. Such structures were never observed on slices of non electrotransferred muscles. However, these structures were not frequent and probably lesional. The surprising fact is that despite the amount of intact plasmid having been greatly reduced between 5 min and 3 h after injection, the level of transfection remains unchanged whether electric pulses were delivered 20 s or 3 h after injection. Such a behavior was similarly observed when injecting 0.3, 3 or 30 microg of plasmid DNA. Moreover, the transfection level was correlated to the amount of plasmid DNA injected. These results suggest that as soon as it is injected, plasmid DNA is proportionally partitioned between at least two compartments. While a major part of plasmid DNA is rapidly cleared and degraded, the electrotransferable pool of plasmid DNA represents a very small part of the amount injected and belongs to another compartment where it is protected from endogenous DNAses. PMID- 14746909 TI - Analysis of the human hexokinase II promoter in vivo: lack of insulin response within 4.0 kb. AB - To locate regulatory element(s) that mediate(s) the effect of insulin on hexokinase II (HKII) gene transcription, we have generated several transgenic mouse lines harboring 97, 254, 505, 819 or 4077 bp of the proximal promoter of the human HKII gene driving the expression of a luciferase reporter gene. Luciferase activities indicate that major promoter elements responsible for the basal activity and tissue-specificity of the human HKII gene expression are located within the 505-bp segment of the promoter. To induce the promoter constructs by endogenous insulin released from the pancreatic beta-cells, transgenic mice were given repeated intraperitoneal injections of D-glucose. Significant induction of luciferase activity was not observed in any of the transgenic mouse lines even though the endogenous HKII mRNA was induced 2.7-fold upon treatment. Thus, our results suggest a lack of insulin response in the 4.0 kb region of the proximal promoter of the human HKII gene in mice. PMID- 14746910 TI - Calfection: a novel gene transfer method for suspension cells. AB - We have developed a novel method called Calfection for gene delivery to and protein expression from suspension-cultivated mammalian cells. Plasmid DNA was simply diluted into a calcium chloride solution and then added to the cell culture for transfection. We evaluated and optimized this approach using suspension-adapted HEK293 cells grown in 12-well plates that were shaken on an orbital shaker. Highest expression levels were obtained when cells were transfected at a density of 5x10(5) cells/ml in the presence of 9 mM calcium and 5 microg/ml of plasmid DNA while maintaining a culture pH of 7.6 at the time of transfection. Suspension-adapted BHK 21 and CHO DG 44 cells could also be transfected using this method. Calfection differs from the widely known calcium phosphate coprecipitation technique. The physico-chemical composition of the DNA interacting complexes is not yet known. The transfection cocktail, DNA in a calcium chloride solution, remained highly efficient during long-term storage at temperatures ranging from room temperature to -80 degrees C. In contrast, calcium phosphate-DNA cocktails are only efficient for gene transfer when prepared fresh. Furthermore, passing the calcium-plasmid DNA mixture through a 0.2-microm filter did not compromise protein expression, whereas calcium phosphate-DNA coprecipitates were retained by the filter. High protein expression levels, a limited number of manipulations and the possibility to filter the cocktail make the Calfection approach suitable for both large-scale transfection in bioreactors and for high-throughput transfection experiments in microtiter plates. PMID- 14746911 TI - Common carp metallothionein-1 gene: cDNA cloning, gene structure and expression studies. AB - Metallothionein-1 (MT-1) cDNA clones were isolated from a common carp (Cyprinus carpio) uninduced hepatopancreas cDNA library. Northern blot assay using the common carp (cc) MT-1 cDNA as a probe showed high fold induction of ccMT mRNA levels in the intestine and kidney following exposure to Cd2+ and Zn2+. Using polymerase chain reaction (PCR), primers designed from the cDNA sequences allowed the isolation of ccMT-1 gene fragments including the 5'-flanking region. The 600 bp 5'-flanking region of ccMT-1 gene carries four putative metal regulatory regions, one AP1, two SP1, one c-Jun site, and a TATA box. The 5'-flanking region of the ccMT-1 gene obtained was a functional promoter responding to the administration of various metal ions as well as hydrogen peroxide (H2O2) and lipopolysaccharide (LPS). When tested in primary cultures of cc hepatocytes, Zn2+ had the highest fold (20 times) induction of the 600 bp cloned ccMT-1 gene promoter, followed by Cu2+, Hg2+, Ni2+ and Pb2+ (4-5-fold inductions); H2O2 and LPS had a 6-7-fold induction. In conclusion, the ccMT-1 is a constitutively expressed MT and its gene promoter is inducible by various metal ions and chemical agents. PMID- 14746912 TI - Cloning and expression of the enolase gene from Desulfovibrio vulgaris (Miyazaki F). AB - The gene encoding an enolase from Desulfovibrio vulgaris (Miyazaki F) was cloned and overexpressed in Escherichia coli. A 2.1-kb DNA fragment, isolated from D. vulgaris (Miyazaki F) by double digestion with PstI and BamHI, contained an enolase gene (eno) and part of the methylenetetrahydrofolate dehydrogenase gene (folD). The nucleotide sequence of eno indicates that the protein monomer is composed of 434 amino acids. An expression system for eno under control of the T7 promoter was constructed in E. coli. The purified His-tagged enolase formed a homooctamer and was active in the formation of phosphoenolpyruvate (PEP) as well as in the reverse reaction, the formation of D-(+)-2-phosphoglyceric acid (2 PGA). The pH dependence and kinetic properties of the recombinant enolase from the sulfate-reducing bacterium were also studied. The amounts of eno mRNA when the bacterium was grown on glycerol or glucose were compared to that when D. vulgaris was grown on lactate. PMID- 14746913 TI - Molecular characterization of a light-responsive gene, breast basic conserved protein 1 (OsiBBC1), encoding nuclear-localized protein homologous to ribosomal protein L13 from Oryza sativa indica. AB - Rice (Oryza sativa L. subsp. indica) cDNA for the gene OsiBBC1, encoding homologue of the breast basic conserved protein 1 (BBC1), similar to ribosomal protein L13, has been identified and characterized. OsiBBC1 codes for a 24 kDa highly basic protein with two potential bipartite nuclear localization signals (NLS) and a transcriptional activation domain (TAD). The structural part of the gene is interrupted by four introns. The OsiBBC1 gene is represented by two copies in the rice genome and both of them are expressed. Northern analysis showed that OsiBBC1 is expressed more in the young root, post-fertilized influorescence, leaf base and callus tissue, which are comprised of actively dividing cells, indicating its role in cell division. The OsiBBC1 transcript accumulated more in the root of light-grown seedlings as compared to the shoot while its levels were higher in the shoot as compared to root of the etiolated seedlings, indicating its down-regulation by light. The western analysis, carried out using antibodies raised against a recombinant fusion protein, 6xHis-OsiBBC1, corroborated its tissue-specific expression profile observed by northern analysis. In addition, OsiBBC1/RPL13 protein could be targetted to the nucleus by particle bombardment of OsiBBC1::GUS fusion construct in the onion epidermal cells. PMID- 14746914 TI - Cloning and characterization of the promoter region of the human prostaglandin F2alpha receptor gene. AB - The promoter region of the human prostaglandin F2alpha receptor (FP) gene was isolated, sequenced, and characterized. The 5'-flanking region has minimal homology to the bovine, mouse, and rat FP promoters with the exception of a 100 bp region. The human promoter similarly lacks a canonical TATA-box and a CAAT box. Potential binding sites for SP-1, GATA-1, STAT-1, and AP-1 are present in the 5'-flanking region. One major transcription start site was identified using 5' RLM-RACE analysis and mapped to an adenine residue 262 nucleotides upstream from the initiator codon in exon 2. Transfection of HeLa cells with FP promoter GFP deletion constructs indicates that the -2437/-1946 region contains repressor activity. DNase I footprinting analysis of this region identifies a footprint over the GATA-like site at -2400. This suggests repression of basal FP transcription may be mediated by a GATA binding site. PMID- 14746915 TI - Identification and functional analysis of a novel human KRAB/C2H2 zinc finger gene ZNF300. AB - A novel human zinc finger gene, ZNF300, was isolated from human embryos on the base of the enrichment of C2H2-specific mRNAs. The ZNF300 cDNA is 3104 bp long and encodes a 604-amino acid protein with an amino-terminal KRAB domain, and 12 carboxyl-terminal C2H2 zinc finger motifs. According to the genome database, ZNF300 is assigned to chromosome 5q33.1. Northern blot analysis revealed that the ZNF300 transcripts are expressed mostly in heart, skeletal muscle and brain. ZNF300 is localized to the nucleus and the KRAB domain of ZNF300 protein exhibits transcription repressor activity. These results suggest that ZNF300 is a ubiquitous transcription repressor in the nucleus. PMID- 14746916 TI - Region-specific testosterone modulation of the vasotocin-immunoreactive system in male dark-eyed junco, Junco hyemalis. AB - The nonapeptide vasotocin (VT) is the avian equivalent of the mammalian antidiuretic hormone vasopressin and is believed to control aggressive and reproductive behaviors. Brain VT distribution has been described in several domesticated avian species. We previously demonstrated that VT distribution in the brain of a free-ranging male passerine, the dark-eyed Junco, Junco hyemalis, resembles that in domesticated birds. A preliminary study also suggested that the VT-immunoreactive (VT-ir) system of juncos is regulated by testosterone (T), as is the case of galliforms. To test this hypothesis, we investigated the effects of castration and T replacement on brain VT-ir innervation in adult male juncos. Castration reduced VT-ir innervation in the lateral septum (SL), the medial preoptic nucleus, the nucleus of the stria terminalis and the intercollicularis nucleus. These effects of castration were largely reversed by T treatment at high physiological doses, but significantly so only for the SL. Given the demonstrated behavioral role of the above VT-ir-containing brain regions, the results suggest that these regions may be sites of action of VT on reproductive behaviors. PMID- 14746917 TI - The epithalamus of the developing and adult frog: calretinin expression and habenular asymmetry in Rana esculenta. AB - Expression of the calcium binding protein (CaBP) calretinin (CR) was studied with immunohistochemistry in the pineal complex and habenular nuclei (HN) of the developing and adult frog Rana esculenta. The frog pineal complex is a medial structure formed by two interconnected components, the frontal organ and the pineal organ or epiphysis; the habenular nuclei are bilateral and are asymmetric due to subdivision of the left dorsal nucleus into medial and lateral components. In the pineal complex, calretinin immunostaining of cells and fibers was consistently observed in developing and adult frogs. In the habenulae, calretinin immunoreactivity exhibited instead marked variations during development, and was expressed only in cells of the medial subnucleus of the left dorsal habenula. In particular, calretinin was detected at larval stages, peaked during metamorphosis, was markedly downregulated at the end of metamorphosis, and was evident again in adulthood. This sequence of calretinin expression was confirmed by quantitative analysis of immunoreactive cells in the left habenula. In tadpoles, calretinin-positive cells exhibited a dorsoventral gradient of density, while in adulthood, they were distributed throughout the dorsoventral extent of the medial subnucleus. The study demonstrates a peculiar developmental pattern, with transient downregulation, of asymmetric calretinin expression in the frog epithalamus. The findings indicate that calcium and calcium buffering systems may play critical roles in neurogenetic and neuronal migration processes implicated in the formation of the asymmetric habenular portion in amphibians. In addition, the reappearance of calretinin expression in the adult frog supports a distinct functional role of the asymmetric habenular component in amphibians. PMID- 14746919 TI - Anoxia induces matrix shrinkage accompanied by an increase in light scattering in isolated brain mitochondria. AB - It is important to monitor mitochondrial conditions, and light scattering (LS) measurements have been applied to the detection of morphological changes in mitochondria in vivo. Little is known about the morphological and LS responses of brain mitochondria to oxygen withdrawal, a critical factor in cell death. We have therefore investigated the morphological and LS responses of isolated brain mitochondria to anoxia. Anoxia induced an increase in LS, reflecting mitochondrial matrix shrinkage. This response was reversible, but was reduced by adding digitonin, which disrupted the outer membrane selectively. This suggested that integrity of the outer membrane was necessary for the matrix response. We further examined the effects of Mg2+ and ATP on the responses because both exist in cells and modulate the changes in matrix volume. Although Mg2+ and ATP reduced the rates of increase and decrease in LS, respectively, the magnitudes of the increases in LS caused by anoxia stayed at over 80% of the control level (no Mg2+) in the presence of Mg2+ and ATP. This suggested that the increase in LS occurred in cells containing Mg2+ and ATP during anoxia. In contrast, that caused by inhibitors of the electron transport chain was reduced to below 30% of the control level in the presence of Mg2+. The present in vitro study provides a basis for interpretation of LS signals from mitochondria in brain research during oxygen withdrawal. PMID- 14746918 TI - A chemical LTP induced by co-activation of metabotropic and N-methyl-D-aspartate glutamate receptors in hippocampal CA1 neurons. AB - In CA1 neurons of guinea pig hippocampal slices, long-term depression (LTD) was induced in the field EPSP response in the absence of test synaptic inputs (one stimulus every 20 s) by application of the metabotropic glutamate receptor (mGluR) agonist, aminocyclopentane-1S, 3R-dicarboxylic acid (ACPD). This effect was blocked and long-term potentiation (LTP) was induced by co-application of N methyl-D-aspartate (NMDA) during ACPD perfusion (ACPD/NMDA-induced LTD). These results indicate that the state of NMDA receptor activation during ACPD perfusion determines whether LTP or LTD is induced in hippocampal CA1 neurons. Co application of an inositol 1, 4, 5-trisphosphate (IP3) receptor inhibitor, 2 aminotheoxydiphenyl borate, during ACPD application had no effect on the ACPD/NMDA-induced LTP, but increased the magnitude of the ACPD-induced LTD, suggesting that the ACPD/NMDA-induced LTP involves NMDA receptors, but not IP3 receptors, whereas the converse applies to the ACPD-induced LTD. PMID- 14746920 TI - Paced mating behavior in the female rat following lesions of three regions responsive to vaginocervical stimulation. AB - The present study examines the effects of ibotenic acid lesions of the medial amygdala, the bed nucleus of the stria terminalis and the medial preoptic area on the display of paced mating behavior in female rats. Lesions of either the medial amygdala or the bed nucleus of the stria terminalis have no effect on the display of paced mating behaviors in ovariectomized, hormone-primed rats. In contrast, lesions of the medial preoptic area significantly lengthen contact-return latencies following intromissions and ejaculations and increase withdrawal from the male following intromissions. The present study demonstrates that the medial amygdala and the bed nucleus of the stria terminalis are not involved in the behavioral responses accompanying paced mating behavior, whereas the medial preoptic area is a critical component of the neural circuit mediating paced mating behavior as well as other appetitive aspects of mating. PMID- 14746921 TI - Nilvadipine antagonizes both Abeta vasoactivity in isolated arteries, and the reduced cerebral blood flow in APPsw transgenic mice. AB - The development of Alzheimer's disease (AD) is generally thought to correlate with cerebral accumulation of Abeta. It has previously been shown that Abeta peptides enhance vasoconstriction in isolated arteries and oppose certain vasorelaxants. Moreover, exogenous application of Abeta peptides causes cerebral vasoconstriction in rodents and in transgenic mouse models of AD that overexpress Abeta there is reduced cerebral blood flow. In the present study, we investigated the effect of nilvadipine, a dihydropyridine-type calcium channel blocker, on Abeta induced vasoconstriction in isolated arteries and in vivo on cerebral blood flow (CBF) of an AD transgenic mouse model overexpressing Abeta (Tg APPsw line 2576). Nilvadipine completely inhibited the vasoactivity elicited by Abeta in rat aortae and in human middle cerebral arteries. The effect of a short treatment duration (2 weeks) with nilvadipine on regional CBF was investigated in 13-month old Tg APPsw mice and control littermates using a laser Doppler imager. Additionally, CBF was also measured in 20-month-old Tg APPsw mice and control littermates that were chronically treated with nilvadipine for 7 months. Untreated Tg APPsw mice showed a reduction of regional CBF compared to their untreated control littermates. Nilvadipine restored cortical perfusion levels in Tg APPsw to values similar to those observed in control littermates without notably affecting the CBF of control mice. All together, these data suggest that nilvadipine might be useful for the treatment of oligemia associated with AD. PMID- 14746922 TI - Excitatory and inhibitory postsynaptic currents of the superior salivatory nucleus innervating the salivary glands and tongue in the rat. AB - The excitatory and inhibitory synaptic inputs to parasympathetic preganglionic neurons in the superior salivatory (SS) nucleus were investigated in brain slices of neonatal (4-8 days old) rat using the whole-cell patch-clamp technique. The SS neurons innervating the submandibular and sublingual salivary glands and innervating the lingual artery in the anterior region of the tongue were identified by retrograde transport of a fluorescent tracer. Whole-cell currents were evoked by electrical stimulation of tissue surrounding the cell. These evoked postsynaptic currents were completely abolished by antagonists for N methyl-D-aspartate (NMDA) glutamate, non-NMDA glutamate, gamma-aminobutyric acid type A (GABAA), and glycine receptors, suggesting that SS neurons receive glutamatergic excitatory, and GABAergic and glycinergic inhibitory synaptic inputs. In SS neurons for the salivary glands, the ratio of the NMDA component to the total excitatory postsynaptic current (EPSC) was larger than that of the non NMDA component. This profile was reversed in the SS neurons for the tongue. In SS neurons for the salivary glands, the ratio of the GABAA component to the total IPSC was larger than the ratio of the glycine component to total inhibitory postsynaptic current (IPSC). The decay time constants of the GABAA component were slower than those for glycine. These characteristics of the excitatory and inhibitory inputs may be involved in determining the firing properties of the SS neurons innervating the salivary glands and the tongue. PMID- 14746923 TI - Green tea polyphenol (-)-epigallocatechin gallate attenuates the neuronal NADPH d/nNOS expression in the nodose ganglion of acute hypoxic rats. AB - Recent studies have shown that (-)-epigallocatechin gallate (EGCG), one of the green tea polyphenols, has a potent antioxidant property. Nitric oxide (NO) plays an important role in the neuropathogenesis induced by brain ischemia/reperfusion and hypoxia. This study aimed to explore the potential neuroprotective effect of EGCG on the ganglionic neurons of the nodose ganglion (NG) in acute hypoxic rats. Thus, the young adult rats were pretreated with EGCG (10, 25, or 50 mg/kg, i.p.) 30 min before they were exposed to the altitude chamber at 10,000 m with the partial pressure of oxygen set at the level of 0.27 atm (pO2=43 Torr) for 4 h. All the animals examined were allowed to survive for 3, 7, and 14 successive days, respectively, except for those animals sacrificed immediately following hypoxic exposure. Nicotinamide adenine dinucleotide phosphate diaphorase (NADPH d) histochemistry and neuronal nitric oxide synthase (nNOS) immunohistochemistry were carried out to detect the neuronal NADPH-d/nNOS expression in the NG. The present results show a significant increase in the expression of NADPH-d/nNOS reactivity in neurons of the NG at various time intervals following hypoxia. However, the hypoxia-induced increase in NADPH-d/nNOS expression was significantly depressed only in the hypoxic rats treated with high dosages of EGCG (25 or 50 mg/kg). These data suggest that EGCG may attenuate the oxidative stress following acute hypoxia. PMID- 14746924 TI - Molecular anatomy of the cerebral microvessels in the isolated guinea-pig brain. AB - Isolated organ preparations represent valuable models for biomedical research, provided that the functional and morphological integrity of vascular and parenchymal compartments is preserved. In this investigation, we have studied the molecular organization of the cerebral microvessels in the isolated guinea-pig brain maintained in vitro by arterial perfusion, a preparation previously proposed as a model of blood-brain barrier (BBB). Using lectin cytochemistry and immunohistochemistry, we examined the microvasculature of the cerebral cortex after 5 h in vitro to assess: (a) the structure of the endothelial glycocalyx at microscopical and ultrastructural level; (b) the distribution of the junctional molecules occludin, ZO-1, PECAM-1 and vinculin; (c) the distribution of basal lamina molecules, such as collagen type IV, laminin and heparan sulfate proteoglycan. All these components of microvessel wall have been previously shown to be vulnerable to ischemic conditions and their organization could be altered in consequence of the transient hypoxia associated with the brain isolation procedure. Our observations demonstrate that the distribution pattern of the molecules considered (i) is comparable to that shown in the cerebral microvasculature of other mammals and (ii) is similar in brains maintained in vitro and in control brains perfused in situ with fixative. The complex of our observation indicates that the molecular organization of the cerebral microvessels is preserved in isolated guinea-pig brain, thus indicating that these preparations can be used to study the cerebrovascular structure and blood brain barrier function in a variety of experimental conditions. PMID- 14746925 TI - Effects of berberine on potassium currents in acutely isolated CA1 pyramidal neurons of rat hippocampus. AB - The effects of berberine, an isoquinoline alkaloid with antiarrhythmic action, on voltage-dependent potassium currents were studied in acutely isolated CA1 pyramidal neurons of rat hippocampus by using the whole-cell patch-clamp techniques. Berberine blocked transient outward potassium current (IA) and delayed rectifier potassium current (IK) in a concentration-dependent manner with EC50 of 22.94+/-4.96 microM and 10.86+/-1.06 microM, Emax of 67.47+/-4.00% and 67.14+/-1.79%, n of 0.77+/-0.08 and 0.96+/-0.07, respectively. Berberine 30 microM shifted the steady-state activation curve and inactivation curve of IA to more negative potentials, but mainly affected the inactivation kinetics. Berberine 30 microM positively shifted the steady-state activation curve of IK. These results suggested that blockades on K+ currents by berberine are preferential for IK, and contribute to its protective action against ischemic brain damage. PMID- 14746926 TI - Nitric oxide decreases a calcium-activated potassium current via activation of phosphodiesterase 2 in Helix U-cells. AB - In the present study, we investigated the underlaying mechanism of nitric oxide (NO) and cGMP on the decline of a Ca2+-activated potassium (KCa) current in U cells of the right parietal ganglion of the pulmonate snail, Helix pomatia. Using a two-electrode voltage-clamp technique, we activated a KCa-current either by opening of endogenous voltage-gated Ca2+-channels during depolarizing voltage steps or by ionophoretic injection of Ca2+ via a third electrode containing 100 mM Ca2+. KCa-current amplitude in U-cells was sensitive to Ba2+, TEA, iberiotoxin, kaliotoxin and charybdotoxin (ChTX), but not to 4-aminopyridine (4 AP) (up to 30 mM) and apamin (up to 300 nM). Thus, the biophysical and pharmacological profile of the KCa-current in U-cells shares similarities with the large-conductance KCa channel (BKCa). The NO-donor sodium nitroprusside (SNP) or S-nitro-N-acetylpenicillamine (SNAP) as well as NO-gas decreased the KCa current amplitude and decreased the rate of KCa-current activation elicited by Ca2+-injection. Decline of the current amplitude and decrease of activation of KCa-current were qualitatively mimicked by the membrane-permeable cGMP analogue dibutyryl-cGMP (db-cGMP). NO-induced decrease of KCa-current was blocked by methylene blue (50 microM), an inhibitor of the guanylyl-cyclase, and by erytho-9 (2-hydroxyl-3-nonyl) adenine (EHNA) (100 microM), an inhibitor of the cGMP stimulated phosphodiesterase 2 (PDE2). These experiments suggest that the NO mediated decrease of KCa-current in U-cells results from synthesis of cGMP by activation of a guanylyl-cyclase and subsequent activation of PDE2. PMID- 14746927 TI - Bilateral processing of vestibular responses revealed by injecting lidocaine into the eighth cranial nerve in vitro. AB - Extracellular unit responses were recorded from the vestibular nucleus (VN) and medial longitudinal fasciculus during horizontal head rotation of an in vitro turtle brainstem in which the temporal bones remained attached. Units were characterized as type I or type II based on the responses to ipsiversive or contraversive rotation, respectively. Lidocaine injections (0.5-2 microl of 0.5%) into the root of the eighth cranial nerve within the cranium caused rapid effects on unit responses to head rotation. Responses of type I units were reduced by ipsilateral injection but enhanced following contralateral injection. On the other hand, type II units had their responses increased by ipsilateral injections yet decreased by contralateral injections. In approximately half of the type II cells, decrease of the contraversive response was accompanied by the appearance of latent ipsiversive activity. Our findings not only confirm that each eighth nerve has afferents that drive ipsiversive excitation of both vestibular nuclei but also suggest that both nerves compete to dominate a central neuron's vestibular response. These results may be inconsistent with the push-pull vestibular model in which each nerve drives the central neuron with a complementary response that enhances the vestibular output. An alternate model is described in which vestibular neurons receive bilateral excitation, and that excitatory input is antagonized by crossed inhibition during contraversive motion. PMID- 14746928 TI - Selective alterations of brain osmolytes in acute liver failure: protective effect of mild hypothermia. AB - The principal cause of mortality in patients with acute liver failure (ALF) is brain herniation resulting from intracranial hypertension caused by a progressive increase of brain water. In the present study, ex vivo high-resolution 1H-NMR spectroscopy was used to investigate the effects of ALF, with or without superimposed hypothermia, on brain organic osmolyte concentrations in relation to the severity of encephalopathy and brain edema in rats with ALF due to hepatic devascularization. In normothermic ALF rats, glutamine concentrations in frontal cortex increased more than fourfold at precoma stages, i.e. prior to the onset of severe encephalopathy, but showed no further increase at coma stages. In parallel with glutamine accumulation, the brain organic osmolytes myo-inositol and taurine were significantly decreased in frontal cortex to 63% and 67% of control values, respectively, at precoma stages (p<0.01), and to 58% and 67%, respectively, at coma stages of encephalopathy (p<0.01). Hypothermia, which prevented brain edema and encephalopathy in ALF rats, significantly attenuated the depletion of myo inositol and taurine. Brain glutamine concentrations, on the other hand, did not respond to hypothermia. These findings demonstrate that experimental ALF results in selective changes in brain organic osmolytes as a function of the degree of encephalopathy which are associated with brain edema, and provides a further rationale for the continued use of hypothermia in the management of this condition. PMID- 14746929 TI - Defense reaction mediated by NMDA mechanisms in the inferior colliculus is modulated by GABAergic nigro-collicular pathways. AB - Electrical stimulation of the inferior colliculus (IC) causes a behavioral activation together with autonomic responses similar to fear reactions to threatening situations. GABAergic mechanisms exert a tonic inhibitory control on the neural substrates of aversion in the IC insofar as local injections of GABA agonists or antagonists inhibit or mimic these defensive behaviors, respectively. Recently, we have shown that systemic injections of the GABA-A receptor agonist muscimol unexpectedly enhanced the freezing and escape responses provoked by gradual increases in the intensity of the electrical stimulation of the IC. Taking into account that the neural circuits mediated by excitatory amino acids (EAA) in the IC may be responsible for the integration of fear states, in the present study we examined whether the defensive behavior induced by local injections of NMDA into the IC is influenced by prior treatment with systemic muscimol and also whether this GABAergic control could be exerted by GABAergic fibers that project to the inferior colliculus from the substantia nigra pars reticulata (SNpr). Rats were implanted with two guide-cannulae aimed at the IC and SNpr through which drug microinfusions with glass micropipette could be made with reduced brain damage. One week after surgery, the animals received either NMDA (7 nmol/0.2 microl) or saline into the IC and were placed into the middle of an enclosure where behavioral responses such as freezing, crossings, jumping, rearing, and turnings could be measured as an indirect index of unconditioned fear. These animals were pretreated either with saline or muscimol (0.5 mg/kg, IP) or with brain injections of saline or muscimol (1 nmol/0.2 il into SNpr). NMDA applied into the IC produced a behavioral activation with significant increases in all behavioral measures. IP injections of muscimol or into the SNpr enhanced the defense reaction caused by microinjections of NMDA into the IC. These findings give support to the idea that unconditioned defensive responses generated in the IC may be mediated by NMDA mechanisms. Additionally, a reduction of the inhibitory control exerted by nigrocollicular GABAergic neurons seems to be responsible for the unexpected pro-aversive action of systemic injections of muscimol on the neural substrates of aversion mediated by NMDA in the IC. PMID- 14746931 TI - Effects of aging on hyperalgesia and spinal dynorphin expression in rats with peripheral inflammation. AB - The aging process is associated with various morphological and biochemical changes in the nervous system that may affect the processing of noxious inputs. This study showed greater hyperalgesia and up-regulation of spinal dynorphin (DYN) expression in aging than in young adult rats during CFA-induced peripheral inflammation. These data indicate that nociception is regulated differently in aging individuals, a fact that should be considered when selecting treatment strategies for aging populations with persistent pain. PMID- 14746930 TI - Experimental diabetes accelerates accumulation of fluorescent pigments in rat trigeminal neurons. AB - The purpose of this study was to investigate autofluorescent pigment granules (lipofuscin, ceroid) in the trigeminal neurons (TN) during aging and streptozotocin-induced diabetes. Four young adult male rats were injected with streptozotocin (STZ; 50 mg/kg) to produce diabetes (DM), for comparison with four young uninjected control rats and four aged rats (90 weeks old). Eight weeks after STZ injection, all rats were fixed with 4% paraformaldehyde, and paraffin sections of TN were prepared and observed by fluorescence microscopy. We found the number of neurons with autofluorescent pigments had increased to 30.38% of the total in DM compared to 8.98% in the control group, and 25.36% in the aged rats. The area of autofluorescence within those neurons was 16.84% in aged rats, 13.02% in DM and 4.45% in the controls. Thus, DM caused accelerated accumulation of fluorescent granules in trigeminal neurons, and this change may show that premature aging contributes to neuronal functional decline and morphological change. PMID- 14746932 TI - Nicotine attenuates spatial learning deficits induced in the rat by perinatal lead exposure. AB - Maternally lead (Pb)-exposed, juvenile rats exhibit significant deficits in spatial reference memory acquisition and working memory performance in the Morris water maze (MWM). Acute systemic application of nicotine reverses these deficits without affecting behavioral performance of the age-matched, lead-unexposed control animals. These results suggest that nicotinic agonist treatments can ameliorate learning and memory impairments, presumably by compensating for deficient nicotinic function in developmentally lead-exposed animals. PMID- 14746933 TI - Imaging of postthalamic visual fiber tracts by anisotropic diffusion weighted MRI and diffusion tensor imaging: principles and applications. AB - Diffusion weighted MRI offers the possibility to study the course of the cerebral white matter tracts. In the present manuscript, the basics, the technique and the limitations of diffusion tensor imaging and anisotropic diffusion weighted MRI are presented and their applications in various neurological and neurosurgical diseases are discussed with special emphasis on the visual system. A special focus is laid on the combination of fiber tract imaging, anatomical imaging and functional MRI for presurgical planning and intraoperative neuronavigation of lesions near the visual system. PMID- 14746934 TI - Orbital pathology. AB - This overview of orbital pathology deals with different kinds of tumors, inflammatory, vascular, and traumatic diseases, which may involve the orbit. Depending on the respective orbital compartment of the globe, the intrakonal, extrakonal and optic nerve the most important and most frequent lesions are presented with their specific clinical symptoms. Their specific presentation on CT- and MR-imaging is discussed in detail, including the most important differential diagnosis. PMID- 14746935 TI - Intracranial pathology of the visual pathway. AB - Intracranial pathologies involving the visual pathway are manifold. Aligning to anatomy, the most frequent and/or most important extrinsic and intrinsic intracranial lesions are presented. Clinical symptoms and imaging characteristics of lesions of the sellar region are demonstrated in different imaging modalities. The extrinsic lesions mainly consist of pituitary adenomas, meningeomas, craniopharyngeomas and chordomas. In (asymptomatic and symptomatic) aneurysms, different neurological symptoms depend on the location of aneurysms of the circle of Willis. Intrinsic tumors as astrocytoma of any grade, ependymoma and primary CNS-lymphoma require the main pathology in the course of the visual pathway. Vascular and demyelinating diseases complete this overview of intracranial lesions. PMID- 14746936 TI - Soft tissue hemangioma is a comma soft tissue neoplasm. PMID- 14746945 TI - Smoking and female fecundity: the effect and importance of study design. AB - This review evaluates the evidence for a link between smoking and female fertility. Twenty-two primary papers and two reviews were found. All but three indicated a detrimental effect of smoking on reproduction despite varying considerably in their approach, definitions used and populations studied. The strength of association is small but statistically significant and a dose response relationship suggests the theory that smoking is causative. Poor design in particular and the failure to assess confounders weakened many studies and should be addressed in future studies. PMID- 14746946 TI - Antiandrogenic progestins for treatment of signs of androgenisation and hormonal contraception. AB - For treating signs of androgenisation (seborrhoea, acne, hirsutism, alopecia) ethinylestradiol in combination with an antiandrogenic progestin is the treatment of choice in adolescents and females throughout the reproductive years, since these combinations are also highly suitable for contraception. Under long-term treatment a favourable effect on carbohydrate and lipid metabolism can be seen due to the antiandrogenic properties of the progestins. These combinations also correct menstrual cycle disturbances and thus reduce the incidence of anaemia. In addition, the frequency of dysmenorrhoea and pelvic inflammatory disease will be lowered and there is a reduction in the risk of endometrial and ovarian cancer. PMID- 14746947 TI - A small randomised trial of low-dose aspirin in women at high risk of pre eclampsia. AB - OBJECTIVE: To determine if aspirin (ASA) therapy reduces the incidence of pre eclampsia in women at high risk of this condition. STUDY DESIGN: Randomised clinical trial. We recruited pregnant women with gestational age at randomisation <14 weeks, who satisfied the following criteria: chronic hypertension, history of severe pre-eclampsia or eclampsia or intrauterine growth retardation (IUGR) or intrauterine foetal death. Nineteen women in the no-treatment group and 16 in the ASA group were successfully followed up. RESULTS: The mean birthweight was higher in the ASA group than in the no-treatment group (2790 g (S.D. 340 g) versus 2616 g (S.D. 779 g)), but the difference was not statistically significant. We found no statistically significant differences between the groups in the proportion of infants with birthweight below 2500 g (13.3% versus 29.4%) and the number of cases with pregnancy-induced hypertension (PIH)/pre-eclampsia (31.3% versus 36.8%). CONCLUSION: These limited data give some support to the potential favourable effect of early treatment with ASA in pregnant women at risk of PIH and IUGR. PMID- 14746948 TI - Prediction of term birth weight in Hispanic women using an equation based on maternal characteristics. AB - OBJECTIVE: To test the accuracy of a birth weight prediction equation based on maternal characteristics in Hispanic women. STUDY DESIGN: A previously published birth weight prediction equation based on maternal and pregnancy-specific characteristics was used to predict birth weights for 283 normal Caucasian women of Hispanic ethnicity. The accuracy of these birth weight estimates was compared to a similarly selected group of 233 non-Hispanic Caucasian gravidas who were located 2500 miles away. RESULTS: Birth weight predictions for Hispanic women were accurate to within +/- 8.4% (+/- 287 g). The percentages of birth weights predicted to within +/- 15 and +/- 10% of actual values were 84 and 68%, respectively. These results were no different than those that were obtained for non-Hispanic Caucasian gravidas. CONCLUSIONS: A previously derived birth weight prediction equation based on maternal and pregnancy-specific characteristics alone successfully predicts term birth weight in normal Hispanic gravidas to within +/- 8.4% (+/- 287 g). PMID- 14746949 TI - 24 hour rhythm in the timing of pre-labour spontaneous rupture of membranes at term. AB - OBJECTIVE: To study the timing of pre-labour spontaneous rupture of membranes (SROM) in term pregnancies. DESIGN: Prospective cohort. SETTING: A maternity hospital in the United Kingdom. SAMPLE: Women who were more than 37 weeks gestation with confirmed spontaneous rupture of membranes and not in labour after 4 h. METHODS: Women who were admitted into labour ward with a diagnosis of spontaneous rupture of membranes after 37 weeks of gestation were included. The women's demographic details were recorded and inquiries about whether they had sexual intercourse in the preceding 12 h. The final outcome of their pregnancy was recorded and analysed. MAIN OUTCOME MEASURES: (1) The exact time of spontaneous rupture of membranes, (2) the time of onset of spontaneous labour, (3) delivery details. RESULTS: One hundred and ninety-six women were studied. A 24 h rhythm in the timing of spontaneous rupture of membranes was found with 33.2% occurring between 00:00 and 04:00 h. When contractions representing the onset of labour occurred there was no diurnal rhythm to the timing of onset of contractions. CONCLUSIONS: There is a 24 h rhythm in the timing of spontaneous rupture of membranes in term gestations. The physiological reasons for this rhythm are not understood at the present time. PMID- 14746950 TI - Clinical observation of blood loss reduced by tranexamic acid during and after caesarian section: a multi-center, randomized trial. AB - OBJECTIVES: To explore the efficacy and safety of tranexamic acid at caesarian section (CS). STUDY DESIGN: Prospective, randomized, case-controlled clinical trial. POPULATION: One hundred and eighty primiparas were randomized into two groups. The study group, 91 women, received tranexamic acid immediately before CS whereas the control group, 89 women did not. METHOD: Blood was collected during two periods. The first period was from placental delivery to the end of CS and the second was from the end of CS to 2 h postpartum. The quantity of blood was measured and compared between the two groups. Complete blood count, urinalysis, liver and renal function, prothrombin time and activity, were tested in the two groups. RESULTS: Tranexamic acid significantly reduced the quantity of blood from the end of CS to 2 h postpartum: 42.75 +/- 40.45 ml in the study group versus 73.98 +/- 77.09 ml in the control group (P=0.001). It also significantly reduced the quantity of total blood from placental delivery to 2 h postpartum: 351.57 +/- 148.20 ml in the study group, 439.36 +/- 191.48 ml in the control group (P=0.002). No complications or side effects were reported in either group. CONCLUSIONS: Tranexamic acid statistically reduces the extent of bleeding from placental delivery to 2 h postpartum and its use was not associated with any side effects or complications. Thus, tranexamic acid can be used safely and effectively to reduce bleeding resulting from CS. PMID- 14746952 TI - Ethnic differences in the association of factor V Leiden mutation and the C677T methylenetetrahydrofolate reductase gene polymorphism with preeclampsia. AB - OBJECTIVE: This case-control study evaluates the association of the factor V Leiden mutation with preeclampsia and potential synergistic effects of the MTHFR 677T and factor V Leiden mutations with regard to disease risk in two different ethnic populations. STUDY DESIGN: 198 women and their 143 newborns from Germany/Croatia and Indonesia with normal pregnancy or preeclampsia participated in the study. The factor V Leiden mutation was determined by direct sequencing and the MTHFR genotype by a PCR-based RFLP method. RESULTS: The factor V Leiden mutation is rare in Indonesians. In Germans/Croatians, the frequency of the mutation was significantly increased in mothers with preeclampsia compared to controls. No disease association was found for combined factor V Leiden/MTHFR 677T genotypes on the maternal and fetal level. CONCLUSIONS: Our results underline the need for a clear distinction of ethnicity in association studies of functional gene polymorphisms. They further support the concept of preeclampsia as a complex disease with variable contributions of disease genes in different ethnic groups. PMID- 14746951 TI - Elective cervical cerclage versus serial ultrasound surveillance of cervical length in a population at high risk for preterm delivery. AB - OBJECTIVE: To compare pregnancy outcome for women at risk of preterm delivery undergoing elective cervical cerclage in the first trimester or serial transvaginal assessment of cervical length with cerclage only if indicated (control cases). DESIGN: A matched case control study. SETTING: Prematurity clinic at two London teaching hospitals. POPULATION: Women at high risk for preterm delivery. METHODS: Cases of elective cervical cerclage were matched for maternal age, ethnic group, previous cervical surgery, previous second trimester loss and early preterm delivery to women undergoing serial ultrasound surveillance of cervical length. Pregnancy outcome data was collected. Data was analysed using Fisher's exact, Mann-Whitney and Student's t-tests. MAIN OUTCOME MEASURES: Gestation at delivery, rate of delivery <24, 24-32 and 32-37 weeks gestation. RESULTS: Thirty-nine cases of elective cervical cerclage were matched to control cases. Both groups were similar for maternal age, ethnic group, previous cervical surgery, previous second trimester loss and early preterm delivery. Cervical cerclage was performed in 14 (36%) of the control cases due to cervical changes. There was no significant difference in median gestation at delivery (266 days versus 260 days P=0.9), number delivering <24 weeks (15% versus 13% P=0.9), at 24-32 weeks (7.5% versus 15% P=0.6) and at 32-37 weeks (15% versus 13% P=0.9). CONCLUSION: Serial transvaginal ultrasound surveillance of cervical length in women at high risk of preterm delivery appears to reduce cerclage rates without compromising pregnancy outcome. A large multicentered randomised trial is required to confirm these findings. PMID- 14746953 TI - Changes in perinatal care and survival in very preterm and extremely preterm infants in The Netherlands between 1983 and 1995. AB - OBJECTIVE: To evaluate changes in obstetrical and neonatal care for very preterm and extremely preterm infants between 1983 and 1995 in The Netherlands and to evaluate the effect of those changes. STUDY DESIGN: Data on all very preterm or VLBW infants from the linked national obstetrical and neonatal databases of 1995 (N=2066) were compared to data on similar infants from a nation-wide study of very preterm infants born in 1983 (N=1338). RESULTS: Obstetrical and neonatal management changed over time, with an increased number of deliveries in tertiary centres (35.7-60.7%), an increase in C-sections (43.7-56.8%) and prolonged artificial ventilation (3.4-9.5%). Survival until discharge increased from 75 to 90% and neonatal morbidity decreased in relative terms. CONCLUSIONS: The short term outcome for these very preterm and extremely preterm infants has improved. Long-term follow-up through to school age and adulthood of preterm infants is needed to investigate the changes in the sequelae of intensive obstetrical and neonatal care. PMID- 14746954 TI - Reliability of the Bishop score before labour induction at term. AB - OBJECTIVE: To evaluate the agreement within three pairs of observers regarding the Bishop score and an informal global evaluation of the cervix (favourable/unfavourable). STUDY DESIGN: We conducted a reliability study of the Bishop score. Three pairs of examiners (A-B, A-C and D-E) performed independently a cervical examination in 156 term pregnant women admitted for labour induction. We calculated the proportion of agreement and the Kappa coefficient. RESULTS: Perfect agreement between two observers for the Bishop score was found in 44 women (28%). Accepting a difference of one point between the observers, agreement increased to 66%. Weighted Kappa coefficients for the Bishop score were 69, 54 and 35% for each pair of observers. Kappa coefficients for the informal evaluation of the cervix were 64, 45 and 46, respectively. CONCLUSION: Agreement between two observers evaluating the cervix is fair to substantial. An informal evaluation of the cervix is as reliable as the Bishop score. PMID- 14746955 TI - Does granulosa cell apoptosis have a role in the etiology of unexplained infertility? AB - OBJECTIVE(S): To compare granulosa cell apoptosis in patients with unexplained infertility and tubal factor. Accelerated granulosa cell apoptosis may be the cause of unexplained infertility. STUDY DESIGN: Setting was IVF-ET Unit of Cerrahpasa Medical Faculty, Istanbul University. GnRH analogs and gonadotropins were used for ovulation induction in patients with unexplained infertility (n=15) and tubal factor (n=15) undergoing in vitro fertilization and embryo transfer (IVF-ET) procedures. Following HCG injection and follicular aspiration, apoptosis of granulosa cells was assessed using the in situ DNA nick end labelling method and apoptosis rate was further determined by flow cytometry. Apoptosis rates were compared between two groups. Mann-Whitney's U-test and Student's t-test were used for statistics. RESULTS: Apoptosis rate was significantly higher in the unexplained infertility group (33.20 +/- 35.62% versus 10.10 +/- 17.23%). CONCLUSION(S): Granulosa cell apoptosis seems to have a role in the etiology of unexplained infertility. PMID- 14746956 TI - Triptorelin versus cetrorelix in intracytoplasmic sperm injection cycles in women with a single ovary. AB - OBJECTIVE: To compare the response to ovarian stimulation with either the long protocol of a GnRH-agonist or the multiple protocol of a GnRH-antagonist, in women with a single ovary who underwent intracytoplasmic sperm injection (ICSI) cycles. STUDY DESIGN: Retrospective study including 75 ICSI cycles from 26 women. Sixty-three cycles were stimulated with triptorelin/hMG or rFSH, whereas 12 cycles were stimulated with cetrorelix/hMG or rFSH. RESULTS: There was not found any statistical significant difference between the two groups regarding the days of stimulation, the number of gonadotropins' ampoules, the peak estradiol levels, the number of aspirated follicles and the number of retrieved oocytes. The fertilization rate, the number of transferred embryos as well as the cumulative embryo score were also similar in both groups. CONCLUSION: The multiple stimulation protocol of cetrorelix is equally effective with the long protocol of triptorelin in the ovarian stimulation of women with a single ovary. PMID- 14746958 TI - The immunological profile of infertile women after repeated IVF failure (preliminary study). AB - OBJECTIVE: The aim of this preliminary study was to estimate the immunological profile of patients after repeated IVF failures. MATERIALS AND METHODS: Seventeen women after repeated IVF failure and 10 non-pregnant women with a history of successful IVF pregnancies were included in the study. We estimated the presence of the auto-antibodies, such as: antinuclear antibodies (ANA), antithyroid antibodies (ATA), antiphospolipid antibodies (APA), antismooth muscle antibodies (ASMA), and antisperm antibodies (ASA). Furthermore, we estimated the percentage of B-1 CD 19+5+ lymphocytes and NK cells using flow cytometry. RESULTS: In the group of patients after IVF failure the percentage of B-1 CD 19+5+ lymphocytes was higher than 1.5% and significantly higher when compared to controls. Three patients after IVF failure had elevated percentages of peripheral blood NK cells. Fourteen infertile patients after IVF failure (82.3%) had at least one abnormal result on autoimmune testing. One patient from the study group had no positive results of immunological tests. CONCLUSION: Our results suggest that immunological alterations may be involved in the etiopathogenesis of unexplained infertility. Furthermore, the results suggest that there is a need for immunological diagnostics in the group of patients with unexplained infertility A greater number of patients is needed for further investigations. PMID- 14746957 TI - Premature ovarian failure and fragile X premutation: a study on 45 women. AB - OBJECTIVE: The aim of this study was to test for the presence of the fragile X (FRAXA) premutation a group of women with early menopause. STUDY DESIGN: 45 women with idiopathic premature ovarian failure (POF), five with a familial and 40 with a sporadic form, were screened for the presence of FRAXA premutation. A control group of 28 women >45 years, with one or more children and no signs of POF, was also studied. RESULTS: We found three cases of fragile X premutations in women all belonging to the group with sporadic POF. CONCLUSION: Our results seems to confirm previous observations on the non random association between POF and FRAXA premutation. PMID- 14746959 TI - Manual vacuum aspiration (MVA) in the management of first trimester pregnancy loss. AB - BACKGROUND: Manual vacuum aspiration is not widely used for the evacuation of retained products of conception in western Europe despite its well-proven success and safety record. Nor is there much information about its use under intravenous (systemic) analgesia or patient-controlled anaesthesia in modern settings. AIM: To evaluate the use of manual vacuum aspiration for the evacuation of retained products of conception under systemic analgesia or patient-controlled anaesthesia in the management of first trimester miscarriages. METHODS: Fifty-eight women with a diagnosis of first trimester miscarriage (42 missed and 16 incomplete miscarriages) were treated with manual vacuum aspiration under systemic analgesia or patient-controlled anaesthesia. Success rates and patient satisfaction and acceptability were recorded. RESULTS: Of the 58 women recruited, 42 underwent the procedure under systemic analgesia and 15 under patient-controlled sedation while 1 woman opted for general anaesthesia. Successful evacuation was achieved in all cases. Both analgesic methods were associated with high levels of patient satisfaction and acceptability. CONCLUSIONS: Manual vacuum aspiration is an option in the management of all first trimester pregnancy losses. Comparisons with other treatment options are indicated. PMID- 14746960 TI - A biomechanical study of the strength of vaginal tissues. Results on 16 post menopausal patients presenting with genital prolapse. AB - AIMS: Measurements of the tensile and bending strength of samples of vaginal tissue collected during corrective surgery of prolapse. MATERIALS AND METHODS: Our measurements were conducted on two samples of vaginal tissue 2 cm x 2 cm collected during surgical correction of prolapse by vaginal route in 16 post menopausal patients. The samples were collected from posterior vaginal fundus, were orientated, and then fixed on a plate holding the edges and allowing the tissue to be stretched over an orifice of 1 cm. The tensile measurements were made using a suture passed over this distance of 1 cm in one of the two samples by recording the strength curve in order to evaluate the force at rupture of the collagen fibres. The second sample was prepared in the same way and a piston of 1 cm diameter was made to penetrate to determine the strength of breakage of the fibres. The pressure and tensile strength curves were recorded up to rupture of the sample, as was the value of the tissue elongation. RESULTS: There was a great variability in the measurements of maximum strength at rupture of the vaginal samples and in the elongation before rupture of the samples. The mean rupture values in tensile tests were 44 and 59 N in bending with extremes of 12 and 130 N. The values of elongation before rupture of a 10 mm sample were 23 mm in tensile tests and 11 mm in bending tests. There was a great variability of results from one patient to another. There was no relation between the values observed and the patient age. There was a statistical relation between the elongation values of the samples and the maximum force before rupture in both the tensile and bending tests. There was also a relation between the measurement of the maximum force at rupture in bending and in tensile tests although there was no such relation in terms of the values of elongation before rupture. DISCUSSION: There is no published reference concerning the strength at rupture or the tensile strength curves for human vaginal tissues. Vaginal tissues are however commonly used as a suspension component in the vast majority of operations for correcting prolapse or urinary incontinence. These suspensions are made by passing a suture through the thickness of the vaginal tissue. The results that we report do however show that these vaginal tissues are very variable in strength from one patient to another. The same finding was made in terms of the elongation values for the vaginal tissue before rupture. The values in bending tests showed that the highest rupture force values and the greatest mean elongation before rupture were lower than in tensile tests. CONCLUSIONS: These findings could explain some failures of these surgical procedures, which are all based on the tensile strength properties. Finally these results could be included in modelling of the reaction of vaginal tissues to the pressure experienced within the vagina. PMID- 14746961 TI - Important anatomical structures used in paravaginal defect repair: cadaveric study. AB - OBJECTIVE: To examine the variations and the anatomical characteristics of the tendinous arch of pelvic fascia (TAPF), the tendinous arch of levator ani (TALA) and the obturator fascia (Ofa) that are important structures in paravaginal defect repair and their relations with important neurovascular structures. STUDY DESIGN: We carried our study on 10 pelvic halves of five female cadavers fixed in 10% formaldehyde. RESULTS: TALA could show a very high location or a low location near to inferior edge of obturator internus. TAPF was not observed in four of the cases. It was examined as a quite weak structure in two of the cases. The location of obturator vessel-nerve bundle could show difference. Obturator artery (OA) and vein sometimes do not course parallel to obturator vein (OV) and make an inclination and extend to the obturator foramen (OF). The distance between TAPF and the pectineal ligament (PL) (Cooper ligament) was measured as 5 cm on average. The distance between TAPF and the entrance of obturator canal was measured as 3.2 cm on average. While the distance of pudendal vessel-nerve bundle from levator ani (LA) at the anterior border of the spine was 0 mm, 2 cm anteriorly it was measured as 4.4 mm on average. CONCLUSION: Since TAPF does not develop in every case, it is not a safe structure to be used in surgery. If TALA develop downward as a variation, it could be difficult to distinguish from TAPF. Since the obturator fascia is a thin membrane, it is not a strong structure for suture placement. The region that is 2 cm in front of the ischial spine (IS) is a dangerous zone for pudendal vessel-nerve bundle. PMID- 14746962 TI - Anterior vaginal wall repair using local anaesthesia. AB - BACKGROUND: The purpose of the present study was to describe the possibility of surgical repair of anterior vaginal prolapse including amputation of the cervix using local anaesthesia. The description was made according to postoperative complication, recurrence rate, influence on urinary incontinence, and satisfaction of the patient. MATERIAL AND METHODS: Eighty-three women were consecutively operated in the anterior wall of the vagina using local anaesthesia and intravenously supplemented with propofol and fentanyl if needed, from January 1998 until September 2000 at a university clinic in Denmark. Operations executed by 16 different surgeons. Seventy patients were operated exclusively by anterior vaginal wall repair, 13 women were operated by other kinds of vaginal repair at the same time. All patients but one were interviewed by telephone. Ten patients reporting a sense of recurrence were called in for clinical examination. RESULTS: The median age was 64.5 years (range 37-85). The median follow-up period was 12.5 months (range 3-35). Median time of the operation was 25 min (range 10-60) including all patients, 20 min (range 10-60) for the patients solely operated in the anterior vaginal wall. The median time spent in hospital was 1 day (range 0 4). One patient was readmitted. Seven patients (8.4%) suffered from complications postoperatively including the patient readmitted, four of these patients were reoperated. By interview 10 women (12%) reported symptoms of prolapse again. By examination two women (2.4%) were diagnosed as recurrence, and were offered a new operation using local anaesthesia. The incontinence patterns before and after the operation are described. Seventy-nine women (95.2%) were totally satisfied with the type of anaesthesia, two women would not choose local anaesthesia for an eventually new operation, one patient did not know, and one patient was unsolved. CONCLUSIONS: The study illustrates an acceptable duration of the operation despite many different surgeons, a need of short time postoperative observation, low postoperative complication rate, low recurrence rate, and a high level of satisfaction reported by the patients. Surgical repair of anterior vaginal prolapse using local anaesthesia is described according to postoperative complication, recurrence rate, influence on urinary incontinence, and satisfaction of the patient. It is concluded that this way of handling anterior vaginal prolapse is suitable for all kinds of patients and in the out-patient clinic. PMID- 14746963 TI - Heterotopic cervical and intrauterine pregnancy in a spontaneous cycle. AB - Early diagnosis and successful management of a case of viable heterotopic cervical and intrauterine pregnancy (IUP) conceived spontaneously is presented. The clinical presentations, treatment modalities and outcome of heterotopic cervical pregnancy (CP) reported in the literature are reviewed. PMID- 14746964 TI - Steroid cell tumour not otherwise specified during pregnancy: a case report and diagnostic work-up for virilisation in a pregnant patient. AB - Steroid cell tumours not otherwise specified are rare ovarian tumours, which can cause foetal and maternal virilisation. This is the first case report that describes a steroid cell tumour not otherwise specified during pregnancy. Differential diagnosis, a diagnostic work-up and treatment are discussed. PMID- 14746965 TI - Severe jaundice in early IVF pregnancy. AB - Jaundice in early pregnancy after in vitro fertilization (IVF) is extremely rare. We report a case of severe jaundice in an IVF treated patient, with a clinical picture similar to intrahepatic cholestasis of pregnancy (ICP). We suggest strategies to prevent similar cases in the future. PMID- 14746966 TI - Acute hepatic failure in pregnancy. AB - Severe liver dysfunction in late pregnancy is an unusual but dramatic event because it can progress very rapidly to fulminating disease and also because two lives, that of the mother and foetus, are involved. We report a descriptive study of a pregnant woman presenting with severe liver dysfunction. PMID- 14746967 TI - Triploid/diploid mosaicism (69XXY/46XX) presenting as severe early onset preeclampsia with a live birth: placental and cytogenetic features. AB - Triploid/diploid mosaicism was diagnosed following karyotyping of an infant with musculo-skeletal abnormalities delivered because of severe preeclampsia. An area of the placenta appeared unusual with histology suggestive of trophoblastic abnormality. The importance of detailed histopathological examination and ploidy and flow cytometry studies where diagnostic uncertainty exists are highlighted. PMID- 14746968 TI - Extraabdominal pelvic foreign body. PMID- 14746970 TI - Infection by Neospora caninum associated with bovine herpesvirus 1 and bovine viral diarrhea virus in cattle from Minas Gerais State, Brazil. AB - Neospora caninum is a canine parasite which is considered a significant cause of bovine abortion. Two cattle herd groups were serologically studied with the objective of studying the prevalence of infection by N. caninum associated with BHV1 and BVDV infections. In group I, 15 dairy herds (476 samples) naturally infected by the three infectious agents were analyzed,. In group II, three dairy herds (100 samples) of cows vaccinated for two viruses were analyzed, in order to determine the infection prevalence by N. caninum. In the first group, an infection prevalence of 12.61, 34 and 28.3% was determined for N. caninum BHV1 and BVDV, respectively. In the second group, a seropositive prevalence of 46, 85 and 76%, respectively, was determined for N. caninum, BVH1 and BVDV. In the first group, the virus and N. caninum had shown in the first group 4.41% positive samples in association with BVH1, 3.15% with BVDV, and 8.41% with BVH1 and BVDV. PMID- 14746971 TI - Humoral and cellular immune responses against Type I cysteine proteinase of Leishmania infantum are higher in asymptomatic than symptomatic dogs selected from a naturally infected population. AB - Canids are natural reservoirs of Leishmania infantum and have been promoted as experimental hosts to decipher the pathogenesis of human visceral leishmaniasis (VL). In this study, the presence of IgG antibodies as well as the presence of mononuclear leukocytes reactive to different cysteine proteinases (CPs) were examined in 13 L. infantum-infected dogs (six with symptoms, seven asymptomatic). Cysteine proteinases which belong to papain-like enzymes known as clan CA are the most studied CPs of parasite protozoa. These molecules are expressed by the intracellular stages of the parasite and could be immunogenic. We studied Type II CP (CPA) and Type I CP (CPB) with its long C-terminal extension (CTE) which could be highly immunogenic. We showed that the level of antibodies reactive to rCPA is low in both symptomatic and asymptomatic dogs. In contrast, when CPB and CTE were used as antigens, the level of total IgG (with IgG2 superior to IgG1) reached higher values in asymptomatic dogs than in dogs with VL. While the peripheral blood mononuclear cell (PBMC) reactivity was significant when cultured in the presence of freezed/thawed (F/T) lysate, it remained low in presence of CP although always higher for PBMC recovered from asymptomatic dogs. We showed the importance of CPB and CTE in particular as a target of immune response and their potential use for serodiagnosis in asymptomatic dogs. PMID- 14746972 TI - Trypanocidal drug resistance in eastern province of Zambia. AB - A survey to investigate resistance to drugs used in the treatment of bovine trypanosomosis was conducted in the eastern province of Zambia between 1996 and 1998. A cross-sectional study was conducted in three districts (Petauke, Katete, Lundazi) at 34 village sampling sites selected at random from villages that had shown greater than 6% prevalence of bovine trypanosomosis during an earlier survey. A longitudinal study was conducted in same three districts over a 1-year period. The study sites were chosen from the cross-sectional study and included eight sites showing high trypanosomosis prevalence and where no control activities were recorded. Use was made of parasitological methods, tests of resistance in cattle and mice and isometamidium-ELISA. Overall mean prevalence of trypanosomosis was 14.4, with 96% of infections caused by Trypanosoma congolense. The remainder was caused by Trypanosoma vivax (2%) and Trypanosoma brucei (2%). Tests in mice showed that of the stabilates collected, 24 (34%) were resistant to only isometamidium chloride, 8 (11.3%) were resistant to only diminazene aceturate, 1 (1.4%) was resistant to both drugs, and 38 (53.5%) were sensitive to both drugs. At least 2 out of 27 stabilates tested in cattle appeared to be resistant to trypanocidal drugs, 1 to isometamidium and 1 to diminazene. Isometamidium could be detected in only 63 (4.1%) of 1526 serum samples from cattle in the study. Only 6 (2.8%) of 212 serum samples from trypanosome-infected cattle had serum levels of the drug above 0.4 ng isometamidium per ml serum which is indicative for drug resistance in the infecting parasite population. Although some drug resistance is apparent, diminazene aceturate and isometamidium chloride can still be expected to be effective as a sanative pair in this area in most cases, since not more than 1 stabilate of 71 investigated showed evidence of resistance to both drugs. PMID- 14746973 TI - The use of metal chelate affinity chromatography on the isolation of Leishmania chagasi promastigote hydrophobic proteinases. AB - In this work, we have assessed the possibility of isolating metalloproteinase fractions from infective Leishmania chagasi promastigotes. Our strategy was the association of the Triton X-114 method with iminodiacetic chromatography enriched with Zn2+. Thus, by using acid conditions, it was possible to isolate two fractions containing two polypeptides, 59 and 63 kDa. The enzymatic activity assay indicated that the two fractions and the two polypeptides had proteinase activities. In addition, it was proposed that those proteinase activities were affected by the presence of 1,10-phenanthroline, a metalloproteinase inhibitor. With this gentle chromatography strategy proposed it is possible to obtain metalloproteinases from L. chagasi in folding that preserve the enzyme activity. This is important for further studies on pathological complications observed in visceral leishmaniasis. PMID- 14746974 TI - The transmission of mixed Trypanosoma brucei brucei/T. congolense infections by tsetse (Glossina morsitans morsitans). AB - Laboratory experiments and field observations clearly show that tsetse flies can be carriers of mixed trypanosome infections. Question remains how easy it is for the tsetse fly to acquire such a mixed infection during the first bloodmeal. This is of particular importance in the epidemiology of Trypanosoma brucei s.l., often a cryptic infection and difficult to transmit to non-teneral tsetse flies. To determine the transmission rate of T. brucei as part of a mixed infection, teneral Glossina morsitans morsitans were fed once on cattle with a mixed (Trypanosoma brucei brucei/Trypanosoma congolense) or single (T. brucei) infection. Of the 140 flies fed on animals with a mixed infection and examined 30 days later, 4 had a metacylic T. brucei infection, 29 a T. congolense infection and 13 a mixed T. brucei/T. congolense infection. There was no significant difference between the transmission rate of T. brucei as a single or as part of a mixed infection. The high proportion of mixed T.b. brucei/T. congolense infections was explained best by a model implying that if a fly is refractory to T. congolense, it is also refractory to T.b. brucei and vice versa. Hence, results suggest that the transmission of T.b. brucei is affected mainly by the vectorial capacity of flies and not by concurrent trypanosome infections in the host. PMID- 14746975 TI - Prevalence of Trypanosoma evansi infection and associated risk factors in camels in eastern Chad. AB - A cross-sectional study was conducted to estimate the prevalence of Trypanosoma evansi infection (Surra) in herds of camels from the eastern area of Chad. The risk factors associated with disease were also identified. From August 1997 to April 1998, a random sample of 2831 camels from 136 herds was selected. Blood samples were collected and examined for the presence of T. evansi using an antibody (card agglutination test-CATT/T. evansi) and a parasite detection test (buffy-coat technique-BCT). Standardized questionnaires with information about the host and management practices were collected and evaluated for their association with seroprevalence (model 1) and parasitological prevalence (model 2) as indications of host sensitivity. In both models, risk factors were selected using ordinary logistic regression (OLR) and herd effect was evaluated using a generalized estimating equations (GEE) model. The apparent prevalence was 5.3% using BCT and 30.5% with CATT. Real prevalence was estimated at 16.9% +/- 1.4 (alpha = 5%). Overall, 27.9% (BCT) and 94.9% (CATT) of the herds had a least one positive animal. Real herd prevalence was estimated at 42.6 +/- 8.3% (alpha = 5%). Camels of the large transhumants had the highest prevalence (estimated to 30.3% +/- 2.5; 62.9 +/- 12.0 in herds). Risk factors associated with seroprevalence were age, ethnic group, length of seasonal migration and longitude of pasture area in the dry season. Risk factors associated with BCT prevalence were age, length of seasonal migration, longitude of pasture area in the dry season, latitude of pasture area in the rainy season and season of sampling. PMID- 14746976 TI - Enhanced survival of rats concurrently infected with Trypanosoma brucei and Strongyloides ratti. AB - The interaction between the blood protozoan parasite, Trypanosoma brucei and the gastrointestinal nematode parasite, Strongyloides ratti was studied in outbred white albino rats. Rats were grouped and given either single infection with T. brucei or S. ratti or concurrently infected with both parasites. Blood parasitaemia and packed cell volume, faecal egg/larva output, adult worm burden and survivability were monitored in order to assess the interactive effects of the infections. All trypanosome-infected rats became parasitaemic within 1 week of infection but surprisingly parasitaemia was higher in the single than concurrently infected group of rats. In addition all animals with single T. brucei infection had died by 14 days after the infection, whereas animals with concurrent infection were still alive by day 28 after the infection when the experiment was terminated. Concurrent infection resulted in significant increase in daily S. ratti egg/larval output in faeces (P < 0.01), but lesser number of adult worms were recovered from the intestine of sacrificed rats on day 8 post infection. Taken together these results suggest that T. brucei and S. ratti interact in a manner that ameliorates their pathogenic effects resulting in a decrease in the level of parasitaemia and intestinal worm burden and in increased life span of the infected rats. These results differ from the classical immunosuppressive attributes of T. brucei and the results are discussed in the context of the possible immune responses that might have contributed to this outcome and the potential significance of the findings in alternative control method of trypanosomosis. PMID- 14746977 TI - Variations in the immune response to natural Schistosoma mattheei infections in calves born to infected mothers. AB - During previous work Schistosoma antibodies and circulating antigens were detected at birth in the serum from some calves born to Schistosoma mattheei infected mothers. The objectives of the present survey were: (1) to investigate the proportion of calves, born to cows infected with S. mattheei, which have specific antibodies and circulating schistosome antigens present in their serum at birth and (2) to investigate whether the presence or absence of these specific antibodies and/or circulating antigens at birth may affect the pattern of a natural S. mattheei infection in calves from 4 to 5 months of age, when the colostral antibodies are thought to be of negligible importance. A total of 28 calves born to infected mothers were randomly selected. Faeces, serum and colostrum samples were collected from the cows at calving, serum samples were collected from the calves at birth (day 0), after intake of colostrum (day 1) and monthly thereafter up to the age of 10 months. Both serum and colostrum samples were analysed for IgG(H+L) against SWAP mattheei and schistosome circulating anodic antigen (CAA) levels. The calves were exposed to a natural challenge from the age of 4-5 months. Faecal samples were collected from the calves monthly, starting at an age of 5 months up to 10 months, and were examined for faecal egg counts. Nine (group 1) out of the 28 calves were found to have specific antibodies in their serum at birth, in 5 of them CAA levels were also detected. In the other 19 calves (group 2) no IgG(H+L) or CAA were detected. At the end of the study faecal egg counts and CAA levels were significantly lower in calves from group 1 compared to group 2. Results confirm earlier work that specific antibodies and circulating antigens may be present in serum from calves at birth, and show that these calves have lower faecal egg counts and CAA levels after exposure to a natural challenge. PMID- 14746978 TI - Detection of circulating 54 kDa antigen in sera of bovine calves experimentally infected with F. gigantica. AB - The antibody response and circulating antigen levels in bovine calves, infected experimentally with Fasciola gigantica, were monitored using enzyme-linked immunoelectrotransfer blot (EITB) and sandwich ELISA, respectively. By EITB, the infected calves' sera recognized the polypeptides in the range of 54-58 kDa as early as 2 weeks post-infection. By 12th week post-infection, the lower two polypeptides of 12 and 8 kDa had disappeared. In sandwich ELISA, the circulating 54 kDa and whole worm antigen of F. gigantica were detected in the sera samples of infected calves as early as 2 weeks post-infection and persisted until the end of experiment (26th week PI). The 54 kDa antigen of F. gigantica appears to be specific and possesses promising immunodiagnostic potential for early prepatent diagnosis of bovine fasciolosis. PMID- 14746979 TI - Contractile activity of cultured adult Dirofilaria immitis. AB - A method for long-term maintenance of adult heartworms (HW) in culture for use in contractile activity studies was developed. Culture conditions included Eagle's minimum essential medium (MEM) containing Earle's balanced salt solution and MEM vitamins and supplemented with 10% horse serum, pH 7.6, 37 degrees C, and humidified 5% CO2:95% room air atmosphere. Motility was observed for up to 91 days. Reducing the culture atmosphere from 20% oxygen to 5% oxygen reduced acid production and survival to 28 days or less. Spontaneous contractile activity of adult HW coils (1 cm diameter) was measured using an isometric force displacement transducer system. Activity had an arrhythmic pattern of good magnitude that could be recorded after up to 50 days in culture for male HW and after up to 40 days in culture for female HW. Analyses of contractile activity included determination of its amplitude, frequency, contraction index, and basal tension. Amplitude for males (3.4 +/- 1.2 g) (mean +/- S.D.) was significantly greater (P < 0.02) than that for females (3.0 +/- 1.1 g), whereas frequency for females (8.2 +/- 2.3 min) was significantly greater (P < 0.03) than that for males (7.5 +/- 2.3 min). The contraction index for females was 16.7 +/- 13.7 mm/min and for males, 14.4 +/- 9.0 mm/min. The difference was not significant. The contraction index was based on line integration of the record of contractile activity. Amplitude and frequency of contractile activity for anterior segments (2.5 cm), suspended lengthwise, from cultured adult female HW, were not significantly different from results for coils, but the contraction index (34.5 +/- 33.8) was significantly higher (P < 0.01) indicating that the pattern of activity was more uniform in the segments. An applied basal tension of about 4 g was suitable for the coils, while a suitable basal tension for segments was about 1.5 g. For coils, amplitude, frequency, and contraction index increased significantly (P < 0.02) with increase in basal tension. Coils were not sensitive to changes in ionic, glucose, oxygen, and osmol concentrations, and pH (6.8-7.6) of the bathing solution. A pH of 8.0 markedly increased basal tension and contractile activity. At 20 degrees C contractile activity stopped. These effects of pH and temperature were reversible. The HW appeared resistant to changes in their environment. This is probably related to an efficient cuticular barrier. The culture and recording methods used in this study open improved opportunities for quantitative evaluation of responsiveness of HW to a variety of physiological and pharmacological factors. PMID- 14746980 TI - Spirocercosis-associated esophageal sarcomas in dogs. A retrospective study of 17 cases (1997-2003). AB - Seventeen client-owned dogs diagnosed with spirocercosis-associated esophageal sarcomas were retrospectively reviewed. The most common clinical signs noticed were vomiting and/or regurgitation (94%), lethargy and depression (59%), pyrexia and anorexia (41% each). Leukocytosis (82%) and microcytic hypochromic anemia (30%) were the most common hematological abnormalities. Caudal thoracic masses were demonstrated on survey radiographs of 13/15 of the dogs and thoracic spondylitis was detected in 12/15 dogs. Spirocerca lupi eggs were detected in 2/8 patients and worms were demonstrated on 1/11 at necropsy. Ten cases underwent surgical attempt to remove the tumors. In six of them partial esophagectomy (PE) was performed and all of them survived the immediate postoperative hospitalization. Five of the cases that underwent PE also received chemotherapy after surgery (doxorubicin (Adriamycin, Upjohn)) with an average survival time of 267 days. The histopathological results of the esophageal tumors were osteosarcoma (9), fibrosarcoma (5) and undifferentiated sarcoma (1). In areas endemic to spirocercosis, regurgitation or vomiting in dogs and microcytic hypochromic anemia and neutrophilia warrant ruling out esophageal sarcomas. Proper surgical treatment could prolong the dogs' lifespan for months, and improve their quality of life. PMID- 14746981 TI - Effect of dexamethasone treatment on the immune response of Gulf Coast Native lambs to Haemonchus contortus infection. AB - Neonatal and weaner Gulf Coast Native (Native) lambs were studied to determine whether an immunological basis underlies their natural resistance to Haemonchus contortus infection. Neonatal Native lambs (n = 8) and weaner Native lambs (n = 15) were randomly assigned to a treatment or a control group. Lambs in the treatment group received dexamethasone by intramuscular injection three times a week for 10 weeks (neonatal) and 15 weeks (weaners). All lambs were monitored for fecal egg count (FEC), blood packed cell volume (PCV), and white blood cell differential counts on a weekly basis for the duration of the studies. Neonatal lambs were kept on pasture with their dams and weaner lambs were dewormed at weaning and kept in pens where they received trickle infections. Serum antibody titers to H. contortus whole worm antigen (WWA) were determined using ELISA. Lymphocyte proliferation assays on peripheral blood mononuclear cells (PBMC) were done to assess lymphocyte function. All lambs were vaccinated with killed Brucella abortus strain 19 to assess the effect of dexamethasone treatment on antibody response. All lambs were necropsied at the end of each study to recover the contents of the gastrointestinal tract for nematode enumeration and identification. The results showed that mean FEC and mean PCV of the treatment group was significantly higher and lower, respectively, than in the control group in both neonatal and weaner lambs from weeks 6 and 5, respectively. At necropsy, total nematode count was significantly higher in treatment groups than in the control groups. Serum antibody titers to H. contortus WWA were significantly lower in treated groups than in control groups. Treatment groups showed a consistent depression in lymphocyte percentage being significantly lower from week 6 in both neonatal and weaner lambs. No differences were found in the response of PBMC to mitogen stimulation between the groups. Lambs in the control groups showed strong positive brucellosis card tests and the treatment groups did not. Dexamethasone treatment resulted in depression of the immune response and loss of natural resistance of Native lambs to H. contortus infection. The results of these studies suggest that some aspects of the immune response may underlie the natural resistance of Native sheep to H. contortus infection. PMID- 14746982 TI - Effect of acaricides on the activity of a Boophilus microplus glutathione S transferase. AB - In the present study, we report the effect of several acaricides on the enzyme activity of a Boophilus microplus recombinant glutathione S-transferase (rGST). GST was expressed in Escherichia coli and was purified with glutathione (GSH) affinity column chromatography. The kinetic constants were determined by reacting GST with the substrates 1-chloro-2,4-dinitrobenzene (CDNB) and glutathione. We report the effect of several acaricides on the enzyme activity of rGST. Some acaricides (ethion, amitraz, chlorpyrifos, DDT, cypermethrin, diazinon, ivermectin, deltamethrin and flumethrin) inhibited rGST. Contrarily, coumaphos had an activating effect. Although the accurate mechanisms of the B. microplus resistance to acaricides remain elusive, this work helps in understanding how acaricides can interact with GST. PMID- 14746983 TI - Pharmacokinetic evaluation of four ivermectin generic formulations in calves. AB - The plasma concentration profiles of four randomly chosen ivermectin (IVM) generic formulations (IVM G1-G4) were compared after their subcutaneous (SC) administration to healthy calves. The disposition of other avermectin-type endectocide compounds, doramectin (DRM) and abamectin (ABM), was also assessed in the same pharmacokinetic trial. Forty-two parasite-free Aberdeen Angus male calves were randomly allocated into six treatment groups. Animals in each group (n = 7) received SC treatment (200 microg/kg) with one of the commercially available endectocide formulation used in the trial. Blood samples were taken into heparinised vacutainer tubes from the jugular vein prior to and up to 35 days post-treatment. The recovered plasma was analysed by HPLC with fluorescence detection. Large kinetic differences were observed among the DRM, ABM and IVM formulations under evaluation. The DRM plasma concentration profiles were higher than those measured for ABM and all the IVM generic formulations. The higher and sustained plasma concentrations of DRM accounted for greater area under concentration-time curve (AUC) and longer mean residence time (MRT) values compared to those obtained for both ABM and the IVM generic preparations. The pattern of IVM absorption from the site of subcutaneous administration showed differences among the generic formulations under evaluation. The IVM G2 preparation showed higher peak plasma concentration and AUC values (P < 0.05) compared to those obtained after the administration of the IVM G1 formulation. Longer (P < 0.05) MRT values were obtained after the administration of the IVM G3 compared to other IVM generic preparations. The kinetic behaviour of ABM did not show significant differences with that described for most of the IVM formulations. This study demonstrates that major differences on drug kinetic behaviour may be observed when using different endectocide injectable formulations in cattle. PMID- 14746984 TI - To knockout in 129 or in C57BL/6: that is the question. AB - Traditionally, knockout experiments are performed in ES cells derived from the 129 mouse strain, followed by backcrossing with the more robust C57BL/6 strain. C57BL/6-derived ES cells have only occasionally been used in this process. We compared C57BL/6- with 129-derived ES cells directly and reviewed the literature. We found that, although some steps are less efficient, the advantages of C57BL/6 mice more than compensate for these drawbacks. PMID- 14746985 TI - Comparative analysis of processed pseudogenes in the mouse and human genomes. AB - Pseudogenes are important resources in evolutionary and comparative genomics because they provide molecular records of the ancient genes that existed in the genome millions of years ago. We have systematically identified approximately 5000 processed pseudogenes in the mouse genome, and estimated that approximately 60% are lineage specific, created after the mouse and human diverged. In both mouse and human genomes, similar types of genes give rise to many processed pseudogenes. These tend to be housekeeping genes, which are highly expressed in the germ line. Ribosomal-protein genes, in particular, form the largest sub group. The processed pseudogenes in the mouse occur with a distinctly different chromosomal distribution than LINEs or SINEs - preferentially in GC-poor regions. Finally, the age distribution of mouse-processed pseudogenes closely resembles that of LINEs, in contrast to human, where the age distribution closely follows Alus (SINEs). PMID- 14746986 TI - How prevalent is functional alternative splicing in the human genome? AB - Comparative analyses of ESTs and cDNAs with genomic DNA predict a high frequency of alternative splicing in human genes. However, there is an ongoing debate as to how many of these predicted splice variants are functional and how many are the result of aberrant splicing (or 'noise'). To address this question, we compared alternatively spliced cassette exons that are conserved between human and mouse with EST-predicted cassette exons that are not conserved in the mouse genome. Presumably, conserved exon-skipping events represent functional alternative splicing. We show that conserved (functional) cassette exons possess unique characteristics in size, repeat content and in their influence on the protein. By contrast, most non-conserved cassette exons do not share these characteristics. We conclude that a significant portion of cassette exons evident in EST databases is not functional, and might result from aberrant rather than regulated splicing. PMID- 14746987 TI - Conservation of protein-protein interactions - lessons from ascomycota. AB - Interacting proteins from Saccharomyces cerevisiae are evolutionarily conserved and their likelihood of having an ortholog in other ascomycota species correlates with the number of interaction partners. Moreover, interacting proteins show a clear preference to be conserved as a pair, indicating that nature maintains selection pressure on the interaction links between proteins. The conservation of interacting protein pairs between different organisms does not exhibit any bias with respect to protein functional roles. PMID- 14746988 TI - Homeodomain to hexapeptide or PBC-interaction-domain distance: size apparently matters. PMID- 14746989 TI - Reading the entrails of chickens: molecular timescales of evolution and the illusion of precision. AB - For almost a decade now, a team of molecular evolutionists has produced a plethora of seemingly precise molecular clock estimates for divergence events ranging from the speciation of cats and dogs to lineage separations that might have occurred approximately 4 billion years ago. Because the appearance of accuracy has an irresistible allure, non-specialists frequently treat these estimates as factual. In this article, we show that all of these divergence-time estimates were generated through improper methodology on the basis of a single calibration point that has been unjustly denuded of error. The illusion of precision was achieved mainly through the conversion of statistical estimates (which by definition possess standard errors, ranges and confidence intervals) into errorless numbers. By employing such techniques successively, the time estimates of even the most ancient divergence events were made to look deceptively precise. For example, on the basis of just 15 genes, the arthropod nematode divergence event was 'calculated' to have occurred 1167+/-83 million years ago (i.e. within a 95% confidence interval of approximately 350 million years). Were calibration and derivation uncertainties taken into proper consideration, the 95% confidence interval would have turned out to be at least 40 times larger ( approximately 14.2 billion years). PMID- 14746990 TI - Genomic microarrays in the spotlight. AB - Microarray-based comparative genomic hybridization (array-CGH) has emerged as a revolutionary platform, enabling the high-resolution detection of DNA copy number aberrations. In this article we outline the use and limitations of genomic clones, cDNA clones and PCR products as targets for genomic microarray construction. Furthermore, the applications and future aspects of these arrays for DNA copy number analysis in research and diagnostics, epigenetic profiling and gene annotation are discussed. These recent developments of genomic microarrays mark only the beginning of a new generation of high-resolution and high-throughput tools for genetic analysis. PMID- 14746991 TI - Driving change: the evolution of alternative genetic codes. AB - Pioneering studies in the 1960s that elucidated the genetic code suggested that all extant forms of life use the same genetic code. This early presumption has subsequently been challenged by the discovery of deviations of the universal genetic code in prokaryotes, eukaryotic nuclear genomes and mitochondrial genomes. These studies have revealed that the genetic code is still evolving despite strong negative forces working against the fixation of mutations that result in codon reassignment. Recent data from in vitro, in vivo and in silico comparative genomics studies are revealing significant, previously overlooked links between modified nucleosides in tRNAs, genetic code ambiguity, genome base composition, codon usage and codon reassignment. PMID- 14746992 TI - Corn and humans: recombination and linkage disequilibrium in two genomes of similar size. AB - Two species with genomes of almost identical size, maize and human, have different evolutionary histories, and as a result their genomes differ greatly in their content of retroelements, average size of the genes and amount of genetic diversity. However, there are also significant similarities: they both have undergone bottlenecks during their recent history and seem to have non-uniform distribution of recombination events. The human genome has been shown to contain large linkage blocks characterized by a limited number of haplotypes. A similar linkage block structure is likely to exist in maize. Although highly diverse maize populations show rapid decline of linkage disequilibrium, as in humans, it is possible to define populations with strong linkage disequilibrium, suitable for whole-genome scan association mapping. The genetic diversity and lack of sequence homology found in maize influences recombinational properties and local linkage disequilibrium levels but also challenges our understanding of the relationship between the genome sequence and species definition. PMID- 14746993 TI - Neuromarketing: beyond branding. PMID- 14746994 TI - Marijuana and multiple sclerosis. PMID- 14746995 TI - Thalamic stimulation in essential tremor. PMID- 14746996 TI - Cytokines in sudden infant death syndrome. PMID- 14746997 TI - GENSAT: a genomic resource for neuroscience research. PMID- 14746998 TI - Restless-legs syndrome in primary care: counting patients in Idaho. PMID- 14746999 TI - Immunotherapy for Guillain-Barre syndrome. PMID- 14747000 TI - Unruptured intracranial aneurysms: benign curiosity or ticking bomb? AB - 15 years ago, the treatment of incidentally discovered intracranial aneurysms was straightforward with a good evidence base behind it. When intracranial aneurysms were identified, people were referred to neurosurgeons who would offer surgical repair if the patient was in reasonable health and had a good life expectancy. Since that time, several studies have given contradictory evidence for what should be done with these lesions, and a new technique for the repair of aneurysms, endovascular coil embolisation, has been developed. Here we review the research and make several recommendations. First, incidentally discovered aneurysms in the anterior circulation less than 7 mm in size in people with no personal or family history of subarachnoid haemorrhage should be left untreated. Second, people with remaining life expectancy of less than 20 years or so (ie, those over age 60 years) should be informed that from a statistical point of view the benefits of treatment do not outweigh the risks. Third, in all other cases treatment with surgical clipping or coil embolisation should be advised. And finally, if surgical treatment is not feasible then medical hypotensive treatment may be a viable alternative. PMID- 14747001 TI - Multiple system atrophy. AB - Multiple system atrophy (MSA) is a sporadic neurodegenerative disorder characterised clinically by any combination of parkinsonian, autonomic, cerebellar, or pyramidal signs and pathologically by cell loss, gliosis, and glial cytoplasmic inclusions in several CNS structures. Owing to the recent advances in its molecular pathogenesis, MSA has been firmly established as an alpha-synucleinopathy along with other neurodegenerative diseases. In parallel, the clinical recognition of MSA has improved and the recent consensus diagnostic criteria have been widely established in the research community as well as movement disorders clinics. Although the diagnosis of this disorder is largely based on clinical expertise, several investigations have been proposed in the past decade to assist in early differential diagnosis. Symptomatic therapeutic strategies are still limited; however, several candidate neuroprotective agents have entered phase II and phase III clinical trials. PMID- 14747002 TI - Genetics of multiple sclerosis. AB - Multiple sclerosis (MS) is probably aetiologically heterogeneous. Systematic genetic epidemiological and molecular genetic studies have provided important insights. Both genetic and non-genetic (environment, stochastic) factors may be involved in susceptibility as well as outcome, but we have yet to understand their relative roles. Any environmental factor is likely to be ubiquitous and act on a population-basis rather than within the family microenvironment. Taken together, the results of genome screening studies provide strong evidence for exclusion of a major locus in MS. There are, however, many genes that seem to be associated with MS. These include, but are in no way limited to, HLA classes I and II, T-cell receptor beta, CTLA4, ICAM1, and SH2D2A. The future of MS genetics, as for most common complex disorders, will be dependent on the resources available, ranging from biological samples and comprehensive databases of clinical and epidemiological information to the development of new technologies and statistical methods. PMID- 14747003 TI - Brain stimulation for epilepsy. AB - Neural stimulation is a promising new technology for the treatment of medically intractable seizures. Vagus-nerve stimulation (VNS) is licensed in several countries as an adjunctive therapy. VNS is as effective as antiepileptic drug therapy, and serious complications are rare. Transcranial magnetic stimulation is simple, non-invasive, and widely used in neurophysiology. Therapeutic results in a few studies are equivocal at best. Deep brain stimulation, although experimental, has been applied to the cerebellum, caudate nucleus, centromedian thalamus, anterior thalamus, subthalamus, hippocampus, and neocortical seizure foci. Preliminary results are encouraging, but not conclusive. Electrode implantation in the brain for indications other than seizures has been associated with a 5% risk for intracranial haemorrhage and 5% for infection. A controlled study of anterior thalamic stimulation in patients with intractable partial and secondarily generalised seizures has been started. Future investigations are likely to study extrathalamic sites of stimulation, and effects of stimulation contingent upon detection of or prediction of EEG patterns of epileptiform activity. PMID- 14747004 TI - Lending a helping eye: artists in residence at a memory clinic. AB - An artist-in-residence programme at the Capital Health Memory Clinic in Halifax, Canada, was established 6 years ago. The artists contribute to the clinic's academic mission by helping to describe how Alzheimer's disease treated by cholinesterase inhibitors is providing a better understanding of human cholinergic neurotransmission. The artists also contribute to the clinical programme by helping to establish a therapeutic ambience, and by allowing clinicians to see themselves through their patients' eyes. The artist programme has inspired a design initiative for the improvement of the physical environment for older patients, and has created a unique art collection that is becoming a resource for scholarship. PMID- 14747005 TI - Breaking the diagnosis of dementia. PMID- 14747012 TI - A child's eye view. PMID- 14747013 TI - The Anopheles gambiae genome: an update. AB - As a result of an international collaborative effort, the first draft of the Anopheles gambiae genome sequence and its preliminary annotation were published in October 2002. Since then, the assembly, annotation and means of accession of the An. gambiae genome have been under continuous development. This article reviews progress and considers limitations in the current sequence assembly and gene annotation, as well as approaches to address these problems and outstanding issues that users of the data must bear in mind. PMID- 14747014 TI - Molecular diagnosis of experimental Chagas disease. AB - Diagnostic methods for detecting Trypanosoma cruzi infection are important to allow administration of chemotherapy and as an experimental tool when trying to understand the pathogenesis of Chagas disease. A nested polymerase chain reaction (PCR)-based approach was recently used to demonstrate preferential heart and skeletal muscle tropism in mice of a Mexican T. cruzi isolate. The authors of this study also demonstrated higher sensitivity for this PCR setup compared with a commercially available enzyme-linked immunosorbent assay kit. PMID- 14747015 TI - More plastids in human parasites? AB - Trypanosomatid parasites are disease agents with an extraordinarily broad host range including humans, livestock and plants. Recent work has revealed that trypanosomatids harbour numerous genes sharing apparent common ancestry with plants and/or bacteria. Although there is no evidence of a plastid (chloroplast like organelle) in trypanosomatids, the presence of such genes suggests lateral gene transfer from some photosynthetic organism(s) during trypanosomatid evolution. Remarkably, many products of these horizontally acquired genes now function in the glycosome, a highly modified peroxisome unique to trypanosomatids and their near relatives. PMID- 14747016 TI - Molecular epidemiology pitfalls: some important clarifications. PMID- 14747017 TI - Parasitic protozoa: thiol-based redox metabolism. AB - In a thoughtful and stimulating article, Sylke Muller et al. describe the thiol based redox metabolism of protozoan parasites and believe that the key enzymes involved in this metabolism have potential use as new drug targets. Additional information on this subject is provided here. PMID- 14747018 TI - Annotating the Plasmodium genome and the enigma of the shikimate pathway. AB - The completion of the Plasmodium falciparum genome sequence heralds a new era in the effort to identify all the parasite's genes along with their cellular functions. A combination of bioinformatics and experimental proof will facilitate this process. Many enzymes in metabolic processes have been identified, but several examples exist of incomplete pathways, such as the shikimate pathway. This review uses the example of the shikimate pathway to examine the application of bioinformatics to lead experimental design in post-genomic biology. PMID- 14747019 TI - Malaria immunity in infants: a special case of a general phenomenon? AB - Newborn infants in endemic areas are markedly resistant to Plasmodium falciparum malaria. Consequently, severe disease is rare during the first few months of life, and infections tend to be low density and relatively asymptomatic during this period. Although this is generally ascribed to passively transferred immunity, attempts to identify the targets and mechanisms of this protection have been unsuccessful. The implications of the hypothesis that the progression from resistance through susceptibility and back to resistance during infancy and early childhood reflects the gradual acquisition of IgG to variant surface antigens (VSAs), while protection from maternal VSA-specific IgG steadily fades, are discussed here. PMID- 14747020 TI - Drugs against leishmaniasis: a synergy of technology and partnerships. AB - To date, there are no vaccines against any of the major parasitic diseases, and chemotherapy is the main weapon in our arsenal. There is an urgent need for better drugs against Leishmania. With the completion of the human genome sequence and soon that of Leishmania, for the first time we have the opportunity to identify novel chemotherapeutic treatments. This requires the exploitation of a variety of technologies. The major challenge is to take the process from discovery of drug candidates all the way along the arduous path to the marketplace. A crucial component will be the forging of partnerships between the pharmaceutical industry and publicly funded scientists to ensure that the promise of the current revolution in biology lives up to our hopes and expectations. PMID- 14747021 TI - Wilderness in the city: the urbanization of Echinococcus multilocularis. AB - A distinct increase in fox populations, particularly in urban areas, has been observed in Europe. This is of particular concern in endemic regions of the small fox tapeworm Echinococcus multilocularis, the aetiological agent of human alveolar echinococcosis. Novel tools have facilitated the investigation of the ecology of urban foxes and have demonstrated the urban wildlife cycle of E. multilocularis. Such studies are essential for estimating the risk of transmission to humans and to determine the basics for the development of control strategies. PMID- 14747022 TI - Bayesian statistics for parasitologists. AB - Bayesian statistical methods are increasingly being used in the analysis of parasitological data. Here, the basis of differences between the Bayesian method and the classical or frequentist approach to statistical inference is explained. This is illustrated with practical implications of Bayesian analyses using prevalence estimation of strongyloidiasis and onchocerciasis as two relevant examples. The strongyloidiasis example addresses the problem of parasitological diagnosis in the absence of a gold standard, whereas the onchocerciasis case focuses on the identification of villages warranting priority mass ivermectin treatment. The advantages and challenges faced by users of the Bayesian approach are also discussed and the readers pointed to further directions for a more in depth exploration of the issues raised. We advocate collaboration between parasitologists and Bayesian statisticians as a fruitful and rewarding venture for advancing applied research in parasite epidemiology and the control of parasitic infections. PMID- 14747023 TI - Schistosomiasis control: keep taking the tablets. AB - Despite the limited reports of praziquantel resistance, the relative success of chemotherapy-based control programmes for schistosomiasis has prompted overdue efforts to expand the use of cheap, generic, praziquantel in sub-Saharan Africa. The likely impact of such programmes on the development and spread of praziquantel resistance is uncertain, but this possibility reinforces the need for monitoring the spectrum of praziquantel sensitivity of schistosome populations and for an improved knowledge of the precise targets for the action of the drug. The search for alternatives to praziquantel and other tools for control of schistosomiasis must continue. PMID- 14747024 TI - Pathogenesis and prenatal diagnosis of human cytomegalovirus infection. AB - Congenital human cytomegalovirus (HCMV) infection is the leading infectious cause of mental retardation and sensorineural deafness. Intrauterine transmission and adverse outcome are mainly related to primary maternal infection. Mechanisms of intrauterine transmission are slowly being unraveled and compelling evidence of the importance of using HCMV clinical strains rather than laboratory-adapted strains for in vitro studies is growing. In the absence of a vaccine or a specific antiviral therapy which could be safely administered to pregnant women with primary HCMV infection, the option of prenatal diagnosis has a crucial role in the management of pregnancy complicated by primary HCMV infection. Reliability of prenatal results, however, is still a major concern presenting the risk of either false-negative or false-positive results. However, as more light is shed on the natural history of HCMV infection during pregnancy and fetal life, the predictive value of negative prenatal diagnosis results is becoming more defined, thus improving the quality of counseling. In addition, the availability of different assays for detection of HCMV in both fetal blood and amniotic fluid samples will eventually reduce the risk of false-positive results. Finally, the identification of reliable prognostic markers of fetal disease remains the ultimate goal and a major challenge. PMID- 14747025 TI - Evaluation of a new NASBA assay for the qualitative detection of hepatitis C virus based on the NucliSens Basic Kit reagents. AB - BACKGROUND: Direct detection of HCV RNA by nucleic acid amplification methods is an essential tool in the diagnosis of HCV infections. In-house developed methods based on reverse transcribed polymerase chain reaction (RT-PCR) are widely used but they are laborious and usually lack the standardization required by clinical laboratories. OBJECTIVES: To evaluate the sensitivity and the clinical performance of an HCV specific nucleic acid sequence based amplification (NASBA) assay based on the commercially available, NucliSens Basic Kit (bioMerieux) reagents. STUDY DESIGN: The analytical sensitivity of the Basic Kit-based HCV assay (BK-HCV) was determined using dilutions of the First World Health Organization International Standard for HCV RNA. The performance of the BK-HCV was evaluated at two study sites in comparison with in-house RT-nested PCR (RT nPCR) by testing a total of 77 plasma specimens. Additional HCV laboratory tests such as Amplicor HCV v2.0 (Roche Diagnostics) and genotype were also included in the comparative analysis. RESULTS: The sensitivity of the BK-HCV was 100-150 IU/ml HCV RNA (85-100% hit rate). When evaluating the clinical performance, we found 96-100% correlation between BK-HCV and RT-nPCR, and 85-91% correlation between BK-HCV and Amplicor. The level of efficiency of the BK-HCV for detecting prevalent HCV genotypes was equal to in house RT-nPCR and Amplicor. CONCLUSIONS: The BK-HCV offers adequate sensitivity for diagnostic purposes and equivalent clinical performance to in-house RT-nPCR assays. The BK-HCV could become a suitable alternative to the in-house amplification methods, providing standardized reagents and procedures, plus rapid results to clinical laboratories. PMID- 14747026 TI - Paired measurements of quantitative hepatitis B virus DNA in saliva and serum of chronic hepatitis B patients: implications for saliva as infectious agent. AB - BACKGROUND: After intensive source and contact tracing 20 % of acute Hepatitis B virus (HBV) infections remain unexplained. Saliva may be an unexpected vehicle of HBV DNA transmission. OBJECTIVE: To further explore this hypothesis we evaluated the quantitative levels of HBV DNA in saliva and compared these with the HBV DNA levels measured in serum. STUDY DESIGN: Serum and saliva were collected from 27 chronic HBV patients attending our outpatient clinic. RESULTS: There were 16 men and 11 women; 15 patients were HBeAg positive, anti-HBe negative and 11 patients were HBeAg negative, anti-HBe positive. One patient was HBeAg and anti-HBe negative. Samples of serum and saliva were collected on the same day. All saliva specimens were clear on inspection. HBV DNA in serum was measured by the Digene Hybrid Capture II microplate assay (Digene Diagnostics), the HBV Monitor assay (Roche Diagnostics) as well as an in-house developed HBV DNA TaqMan assay. The HBV DNA TaqMan assay was used for the quantitative measurement of HBV DNA in saliva. Median HBV DNA levels in serum were 2.10 x 10(5) geq/ml and ranged from 373 genome equivalents per ml (geq/ml) to 4.13 x 10(9) geq/ml; median HBV DNA levels in saliva were 2.27 x 10(4) geq/ml and ranged from 373 geq/ml to 9.25 x 10(6) geq/ml. A clear correlation was shown between HBV DNA in serum and saliva; log HBV DNA in saliva=1.01 + 0.56 x (log HBV DNA in serum). CONCLUSIONS: this is the first report of precise quantitative measurements of HBV DNA levels in saliva and the relationship with HBV DNA levels in serum. Our findings show that saliva is a source of HBV DNA. This finding may have implications in selected patients for the infectivity of saliva and offer further insight in the routes of transmission of HBV infection. PMID- 14747027 TI - Molecular variants of HPV type 16 E6 among Mexican women with LSIL and invasive cancer. AB - BACKGROUND: Cervical cancer is the second most common cancer in women worldwide. Infection with human papillomavirus (HPV) 16 is an important risk factor associated with cervical cancer, more than 50% of cervical cancer tissues have DNA of HPV 16. Intratypic variants have been reported, although they differ in prevalence, biological and biochemical properties, their implication in the aetiology of cervical cancer is still uncertain. OBJECTIVE: To identify HPV type 16 E6 variants among Mexican women with diagnosis of low-grade squamous intraepithelial lesion (LSIL) or invasive cancer (IC). STUDY DESIGN: Forty HPV16 positive samples were included, 15 were from women with LSIL, 25 from women with IC; 610 pb from the E6 gene were amplified by PCR and the variant status subsequently determined by hybridization with 27 biotinilated probes. Statistical analysis was performed with chi2, odds ratio (OR). RESULTS: In the LSIL group we only found ten (66%) EP and five (33%) EP350G variants. In the IC group, four variants were found; 11 (44%) AA, seven (28%) EP, six (24%) EP350G, one (4%) Af2. Comparison of the frequency of variants differed from EP in both groups of patients (P=0.01) with an odds ratio (OR) of 5.14 (CI 95% [1.07-26.56]). CONCLUSION: This study demonstrates an association between HPV type 16 variants different from prototype (EP) and invasive cervical cancer. PMID- 14747028 TI - Real-time PCR for rapid diagnosis of entero- and rhinovirus infections using LightCycler. AB - BACKGROUND: PCR techniques have proved to be more sensitive than traditional cell culture in the diagnosis of enterovirus and rhinovirus infections and are widely used in clinical virus laboratories. However, PCR assays are relatively time consuming and labor intensive, particularly if separate hybridization steps are used to confirm the specificity of positive findings. OBJECTIVES: The aim of the present study was to develop fast and sensitive real-time PCR assay, which would allow simultaneous detection of entero- and rhinoviruses and their quantification in clinical and experimental samples. STUDY DESIGN: Two real-time RT-PCR protocols were developed using LightCycler (LC) technology; SYBRGreen and hybridization probe assays. The sensitivity of these assays to detect entero- and rhinoviruses was compared with that of a traditional reference RT-PCR hybridization assay and cell culture. All PCR protocols used the same primers amplifying the 5'-non coding region (NCR) of entero- and rhinoviruses. The LC probe assay and the reference RT-PCR used almost identical detection probes, which bind to enterovirus specific amplicons. RESULTS AND CONCLUSIONS: Both real time PCR assays were equally sensitive as the reference RT-PCR-assay and all were more sensitive than cell culture. Both real-time assays quantified reliably the amount of the virus and took much shorter time than the reference RT-PCR. As the real-time SYBRGreen assay detects both entero- and rhinoviruses it can be used for primary screening of samples, which can be positive for either of these viruses. The real-time probe-assay can confirm the presence of enterovirus in SYBRGreen positive samples or it can be used for selective screening of enteroviruses e.g. from CSF samples. PMID- 14747029 TI - Quality control assessment for the serological diagnosis of dengue virus infections. AB - BACKGROUND: A major drawback of modern society's rapidly increasing mobility is the ease with which dangerous infections can be imported into Europe. Often these infections are not diagnosed because physicians are not familiar with the symptoms and laboratory tests are not always available in local diagnostic centres. Improving diagnostics is the most important step in detecting and dealing with these pathogens and quality control measures are, therefore, essential tools. OBJECTIVES: To assess the diagnosis of imported dengue virus infections in Europe by (1) running a pre-evaluation panel (four serum samples, sent out in 1999) and optimising sample preparation and shipping procedures and (2) initiating an External Quality Assurance (EQA) program (20 serum samples, sent out in 2002). STUDY DESIGN: All serum samples sent out were to be tested for the presence of dengue virus-specific IgM and IgG. For the pre-evaluation panel, four samples were distributed (one sample IgM+/IgG+, one sample IgM-/IgG+, two samples IgM-/IgG-) and for the EQA 20 samples (12 samples IgM+/IgG+, five samples IgM-/lgG+, one sample lgM+/IgG- two samples IgM-/IgG-). 13 laboratories took part in the pre-evaluation panel and 18 laboratories participated in the first EQA run. RESULTS: For the pre-evaluation panel, the participants reported concurrent and correct results for 88% of the IgG-positive samples and for 100% of the IgG negative samples. The results for the IgM-positive sample were correct in 91% of the reported tests and in 97% of the IgM-negative samples. For the EQA, the participants reported concurrent and correct results for 71% of the IgG-positive samples and 89% of the IgG-negative samples. 58% concurrent and correct results were reported for the IgM-positive samples and 97% for the IgM-negative samples. CONCLUSIONS: The results presented here demonstrate the importance of quality measures for imported viral pathogens like dengue viruses and clearly indicate the need for improving the existing test systems. PMID- 14747030 TI - New method of differentiating wild-type varicella-zoster virus (VZV) strains from Oka varicella vaccine strain by VZV ORF 6-based PCR and restriction fragment length polymorphism analysis. AB - A new method was developed to distinguish accurately wild-type varicella-zoster virus (VZV) strains from the Oka vaccine strain. Several DNA fragments covering open reading frame (ORF) 1-37 were amplified from wild-type VZV strains including the Oka parent strain and from the Oka vaccine strain. Restriction fragment length polymorphisms of these regions were compared, and nucleotide differences between the vaccine virus and other wild-type VZV strains were noted in ORFs 6, 10, and 35. In addition, variations of the R2 and R4 reiterated structures of the vaccine and its parent strains were examined. The Oka vaccine strain used in Japan was shown to be a mixture of viruses with different nucleotide sequences that had variations in at least three nucleotide positions in ORF 1-37 and had variable polymorphisms at R2 and R4 repeat regions (two and three patterns, respectively). The Oka parent strain on the other hand showed a single sequence and had only one reiterated structure at these regions. When VZV ORF 6 was amplified and its product was digested with AluI, the Oka vaccine strain could be precisely differentiated from its parent and from 56 other Japanese clinical isolates. PMID- 14747031 TI - Detection of herpes simplex virus type 1, herpes simplex virus type 2 and varicella-zoster virus in skin lesions. Comparison of real-time PCR, nested PCR and virus isolation. AB - BACKGROUND: Herpes simplex virus type 1 (HSV-1), herpes simplex virus type 2 (HSV 2) and varicella-zoster virus (VZV) cause a wide range of signs and symptoms, varying from trivial mucocutaneous lesions to life-threatening infections, especially in immuno-suppressed patients. Since antiviral drugs are available, rapid and sensitive laboratory diagnosis of these virus infections is important. OBJECTIVE: To set up and evaluate HSV-1, HSV-2 and VZV qualitative real-time PCR on the Lightcycler system and to compare the results with those of the 'in-house' nested PCR and virus isolation. STUDY DESIGN: 110 consecutive samples from dermal or genital lesions from patients suspected of having HSV infections and another 110 samples from patients with suspected VZV infections were tested with real time PCR, nested PCR and virus isolation. RESULTS: 24 samples (22%) were positive for HSV-1 by virus isolation and nested PCR, whereas 26 (24%) were positive by real-time PCR. HSV-2 was detected in 28 samples (25%) by virus isolation, in 41 (37%) by nested PCR and in 40 (36%) by real-time PCR. VZV was isolated in 15 samples (14%) and VZV DNA was detected in 51 samples (46%) by nested PCR as well as by real-time PCR. Nucleic acid amplification increased the detection rate of HSV-2 and VZV DNA in particular compared to virus isolation. No significant difference in sensitivity was found between real-time PCR and nested PCR. CONCLUSION: Real-time PCR has the advantage of rapid amplification, a reduced risk for contamination and it is a suitable method for diagnosis of VZV and HSV in specimens from skin lesions. PMID- 14747032 TI - Evaluation of a convenient enzyme immunoassay to assess the quality of genital specimens submitted for the detection of human papillomavirus DNA by consensus PCR. AB - BACKGROUND: In the PGMY-line blot assay, a human beta-globin fragment is co amplified with human papillomavirus (HPV) DNA, and both analytes are detected by hybridization with probes fixed on a strip in a linear array. The beta-globin DIG MWP test also detects beta-globin amplicons, but in a microtiter plate-based enzyme immunoassay format. Although the PGMY-line blot assay detected 50 cells per test, the beta-globin DIG-MWP test generated a signal above the detection cut off with five cells per test. OBJECTIVE: The performance of the beta-globin DIG MWP assay to detect beta-globin DNA was assessed. STUDY DESIGN: The beta-globin DIG-MWP assay was compared to a standard beta-globin PCR and to the PGMY-line blot strips on 401 genital specimens. Overall, the three beta-globin assays were compared on 325 undiluted lysates, 14 diluted lysates and DNA extracted from 62 lysate samples. RESULTS: Concordance between the PGMY-line blot and the standard beta-globin assay reached 99.5% (399 of 401 results), for a kappa value of 0.95. Concordant results were also obtained between the beta-globin DIG-MWP assay and PGMY-line blot assay for 387 (96.5%) of 401 test results, for a kappa value of 0.57. Discordant results were due to the increased sensitivity of the DIG-MWP assay. Using a cut-off for positivity at 1.500 optical density (OD) units for beta-globin DIG-MWP, concordance improved to 100% (401 of 401 results, kappa at 1.00). CONCLUSION: The beta-globin DIG-MWP assay was adequate to screen for sample adequacy for HPV analysis in genital specimens. PMID- 14747033 TI - A case of simultaneous primary HIV-1 and CMV infections. PMID- 14747034 TI - Efficient treatment of paraffin-embedded cervical tissue for HPV DNA testing by HC-II and PCR assays. AB - BACKGROUND AND OBJECTIVES: High-risk human papillomaviruses (HR-HPVs) are the primary cause of cervical cancer. In order to meet with clinical requirements, a direct capture test with signal amplification (HC-II), able to detect the 13 prevalent HR-HPVs, has been developed and validated for cytological specimens. STUDY DESIGN: the use of HC-II assay with formaldehyde-fixed paraffin-embedded cervical biopsies, for retrospective studies or to support histological findings, was investigated by analysing three different sample treatments. The efficacy of this test was compared with a reference PCR-ELISA, using MY09/11 and GP5+/6+ consensus primers and the use of a single extraction method for both HC-II and PCR-ELISA assays was validated. RESULTS: protease treatment of dewaxed biopsy sections allowed an optimal performance of HC-II and has also been validated for PCR-ELISA. Overall, on the analysis of 50 cervical samples HC-II and PCR-ELISA assays showed a high concordance (K=0.80). Compared with PCR-ELISA, the HC-II had a sensitivity of 86.4% and a specificity of 92.9%. The results showed that short amplimers are necessary for a sensitive PCR-ELISA from formaldehyde-fixed paraffin-embedded biopsies, while HC-II showed a relatively low sensitivity for HPV18 within the HR probe pool. CONCLUSIONS: HC-II can be a valid tool for the diagnosis of HPV infection in biopsy material. The possibility to use the same specimen preparation material for both HC-II and PCR-ELISA allows HC-II positive specimens to be further processed by PCR-ELISA if specific genotyping is needed. PMID- 14747037 TI - Prostate-specific antigen doubling time in the identification of patients at risk for progression after treatment and biochemical recurrence for prostate cancer. AB - After primary treatment for clinically localized prostate cancer, biochemical recurrence is usually the first evidence of either local recurrence or metastatic progression. This poses a diagnostic dilemma for both the patient and the physician regarding future therapy. Prostate-specific antigen doubling time (PSADT) is a useful tool in this clinical setting. There have been multiple reports of the utility of PSADT in men with isolated biochemical recurrence after either radical prostatectomy or external-beam radiation therapy. Early observations of PSADT in men with recurrence are reviewed and the current literature is summarized to allow physicians to make an accurate assessment of a patient's risk of progression after isolated biochemical recurrence. PMID- 14747038 TI - Use of nomograms to predict the risk of disease recurrence after definitive local therapy for prostate cancer. AB - The generally indolent nature of prostate cancer, as well as the impact that treatment can have on quality of life (QOL) and cancer control, makes the decision analysis difficult for patients facing the task of selecting a treatment for clinically localized disease. Instruments to aid patients and their physicians in this decision analysis are needed. Nomograms are instruments that predict outcomes using specific clinical parameters. Nomograms use algorithms that incorporate several variables to calculate the predicted probability that a patient will achieve a particular clinical end point. Nomograms tend to outperform both clinical experts and predictive models using methods of risk grouping. We briefly outline the uses and limitations of nomograms, principles of nomogram construction, and the available models for predicting the progression free probability after local definitive therapy with radical prostatectomy, external-beam radiotherapy, or brachytherapy. There is a need for additional nomograms that predict outcomes after salvage therapy, as well other clinical end points, including QOL-adjusted survival. PMID- 14747039 TI - Molecular markers to identify patients at risk for recurrence after primary treatment for prostate cancer. AB - Accurate prognostication is a prerequisite for accurate therapeutics and management of prostate cancer because indolent tumors may require no intervention, whereas aggressive tumors lead to patient mortality. There is a critical need to define these subgroups of patients with prostate cancer differing in clinical outcome. Prognostic nomograms based on clinical data provide useful predictions of clinical states and outcomes, but they need further refinements to improve accuracy and universality. Genomic and proteomic analyses have provided many novel markers that may help define prognostic parameters based on the underlying biology of prostate cancer progression at the molecular level. These molecular markers are likely to augment traditional prognostic modalities by providing a set of molecularly defined and quantifiable variables. Encompassing the genome, transcriptome, and proteome of prostate cancer will likely provide "molecular signatures" that will bridge prognostication, prediction, and treatment in a single continuum. PMID- 14747040 TI - Use of neoadjuvant and adjuvant therapy to prevent or delay recurrence of prostate cancer in patients undergoing radiation treatment for prostate cancer. AB - Multiple oncologic treatment modalities are often integrated into the curative treatment approach for the patient with a newly established diagnosis of cancer. The combination of neoadjuvant and adjuvant therapies with radiotherapy for the care of the patient with prostate cancer is no exception. There is clear evidence that neoadjuvant androgen suppression reduces the volume of tumor in preparation for radiotherapy, and it is an effective addition to conventional-dose external radiotherapy in patients with large-volume primary prostatic tumors. Adjuvant androgen suppression improves local and systemic tumor control and improves survival duration compared with radiotherapy alone for patients with locally advanced or node-positive prostate cancer, particularly in those with high-grade disease. The role of neoadjuvant and adjuvant therapies is under intense scrutiny as several randomized clinical trials seek to optimize the combination of androgen suppression, chemotherapy, and radiotherapy. The historical precedent for combining androgen suppression with radiotherapy is described, as are the results of prior definitive trials and ongoing studies in this setting. PMID- 14747041 TI - Use of neoadjuvant and adjuvant therapy to prevent or delay recurrence of prostate cancer in patients undergoing surgical treatment for prostate cancer. AB - There have been improvements in the outcome of patients with clinically localized prostate cancer treated by radical prostatectomy. However, some patients treated with radical prostatectomy will have clinical or biochemical progression. These men are at increased risk of dying of their disease. Identification of patients with adverse features at the time of radical prostatectomy may permit the use of additional multimodality therapies to improve outcomes. Whether this additional multimodality therapy should be administered in the neoadjuvant or adjuvant setting remains controversial. Further, whether a patient at increased risk for progression after radical prostatectomy requires additional therapy before the development of documented progression remains controversial. This article reviews the potential multimodality approaches to prevent or delay recurrence of prostate cancer in patients undergoing surgical treatment for prostate cancer. PMID- 14747042 TI - Cancer and Leukemia Group B (CALGB) 90203: a randomized phase 3 study of radical prostatectomy alone versus estramustine and docetaxel before radical prostatectomy for patients with high-risk localized disease. AB - The purpose of The Cancer and Leukemia Group B (CALGB) 90203 trial is to determine which of 2 treatment strategies is superior in treating men with high risk, clinically localized adenocarcinoma of the prostate (stage T1 to T3a NX M0), defined as a predicted probability < or =60% of remaining free from disease recurrence for 5 years after surgery. Patients with a > or =10-year life expectancy will be randomized to either radical prostatectomy (RP) alone versus estramustine and docetaxel before RP. Participants will be excluded if they have received prior therapy for prostate cancer (except transurethral resection of the prostate) or are judged not to be appropriate candidates for RP. Eligible patients will be stratified according to their predicted probability of remaining free from disease recurrence at 5 years after surgery (0% to 20%, 21% to 40%, and 41% to 60%) and randomized. Neoadjuvant chemotherapy will be 6 cycles (1 cycle = 21 days) of estramustine (280 mg tid, days 1 to 5) and docetaxel (70 mg/m2 on day 2). Warfarin (2 mg/day orally) will be given for prophylaxis against deep venous thrombosis. Bilateral pelvic lymph node dissection and RP will be performed within 60 days of registration/randomization for men randomized to the surgery alone arm. For men randomized to receive preoperative chemotherapy, the surgical procedure will be performed within 60 days of completion of chemotherapy. Patients will be monitored with history review, physical examination, and serum prostate-specific antigen (PSA) levels every 3 months for the first 3 years after surgery, every 6 months for the next 3 years, and annually thereafter. Biochemical disease recurrence will be defined as a serum PSA level >0.4 ng/mL on 2 consecutive occasions > or =3 months apart after RP. The time of biochemical failure is measured from the date of randomization to the time of the first PSA level <0.4 ng/mL that is confirmed on the second serial PSA. The primary study end point is to determine if early systemic treatment with neoadjuvant estramustine and docetaxel before RP in patients with high-risk prostate cancer will decrease 5-year recurrence rates when compared with RP alone. Secondary outcomes will include (1) the safety and tolerability of neoadjuvant estramustine and docetaxel before RP; (2) the impact of this neoadjuvant strategy on pathologic tumor stage, including lymph node and surgical margin status; (3) time to clinically apparent disease recurrence; and (4) overall survival. The impact of RP with and without neoadjuvant estramustine and docetaxel on the patient's quality of life from pretreatment through year 3 will be assessed. Frozen prostate tissue will be obtained from men undergoing prostatectomy who are enrolled in either the treatment or control arms of the trial. These samples will be analyzed for their RNA expression patterns in order to build outcome prediction models. Furthermore, using array-based methods of expression analysis, the sensitivity to chemotherapeutic agents and response to chemotherapy may likewise be predicted. The trial will enroll approximately 700 men during a 48 month period. Patients will be observed for 84 months after study closure. The power to detect a 36% decrease in 5-year recurrence rates is 90%. PMID- 14747043 TI - Salvage radiotherapy in the treatment of prostate cancer. AB - For patients undergoing radical prostatectomy for prostate adenocarcinoma, the most common cause of failure is an asymptomatic increase in levels of prostate specific antigen (PSA). Salvage radiotherapy (RT) to the prostate bed has been used when there is no clinical evidence of metastatic disease. However, this is still not widely accepted because there is currently no consensus on the optimal management of an isolated PSA failure. Salvage RT given in a select group of patients is effective, with a 70% to 80% biochemical response rate and a long term biochemical control rate as high as 35% to 40%. These data indicate that RT offers a substantial risk of curative salvage of patients who fail radical prostatectomy. Although there is interest in studying investigational modalities (eg, vaccine therapy) among patients with asymptomatic, PSA-detected recurrences after surgery, caution must be applied, and treatment modalities with known curative potential (ie, RT) should be used before noncurative techniques are attempted. This article outlines the rationale, results, and toxicity of salvage RT for an asymptomatic increase in PSA levels, with emphasis on identifying patients with favorable prognostic factors with higher rates of long-term biochemical control with local treatment. PMID- 14747044 TI - Salvage prostatectomy in patients who have failed radiation therapy or cryotherapy as primary treatment for prostate cancer. AB - Asymptomatic prostate-specific antigen (PSA) recurrence after radiation therapy for prostate carcinoma poses a diagnostic and therapeutic dilemma for clinicians. Patients with locally recurrent disease can consider treatment options of salvage surgery, cryotherapy, watchful waiting, or androgen deprivation. Of these options, only salvage surgery has been shown to result in long-term disease-free survival for selected patients. However, salvage surgery is associated with significant morbidity, including urinary incontinence and rectal injuries. Ideally, salvage surgery outcomes can be optimized with careful patient selection according to clinical stage, serum PSA levels before radiation and surgery, the medical condition of the patient, and clear expectations of the physician and patient. Among patients with locally recurrent disease, those with localized prostate carcinoma amenable to radical prostatectomy before radiation or cryotherapy would be the most suitable candidates for salvage surgery. PMID- 14747045 TI - Hormonal therapy options for patients with a rising prostate-specific antigen level after primary treatment for prostate cancer. AB - At this time, many treatment options exist for primary androgen-deprivation therapy. "Standard" options with long-term results on outcome include orchiectomy, monotherapy with a luteinizing hormone-releasing hormone (LHRH) agonist, and combined androgen blockade using an LHRH agonist and antiandrogen. All treatments are associated with some morbidity related to the reduction of circulating testosterone. For this reason, "alternative" regimens are under active study to determine whether equal outcomes can be achieved with lesser toxicity. PMID- 14747046 TI - Role of secondary hormonal therapy in the management of recurrent prostate cancer. AB - Androgen ablation remains the cornerstone of the systemic management of prostate cancer. After initial androgen deprivation, clinical outcomes vary considerably. For the patient with progressive disease after androgen deprivation, multiple therapeutic options are available and include antiandrogen withdrawal, chemotherapy, and secondary hormonal agents. Multiple secondary hormonal agents have clinical activity and the sequential use of these agents may lead to prolonged periods of clinical response. In addition to the use of oral antiandrogens, active secondary hormonal therapies include adrenolytic agents such as ketoconazole and aminoglutethimide, corticosteroids and estrogenic compounds. This article reviews the clinical trial data for these various agents and discusses their role in the management of patients with advanced prostate cancer. PMID- 14747047 TI - Radiation Therapy Oncology Group P-0014: a phase 3 randomized study of patients with high-risk hormone-naive prostate cancer: androgen blockade with 4 cycles of immediate chemotherapy versus androgen blockade with delayed chemotherapy. AB - Currently, approximately 30,000 men die annually of metastatic, hormone refractory prostate cancer. Androgen blockade is palliative and is generally effective for an average of 2 to 3 years until a patient develops androgen independent disease. Newer chemotherapeutic regimens can induce remissions in approximately 50% of patients; however, median survival for patients with androgen-independent disease is still 8 to 12 months. The strategy of using chemotherapy regimens after androgen blockade has been proved noncurative, and new approaches are needed to attempt to cure patients with advanced disease. It has been demonstrated in the preclinical setting that androgen withdrawal induces apoptosis in cancer cells in both the Shinogi breast cancer model and the LNCaP prostate cancer model. In both of these models, androgen withdrawal was not curative, and the tumors grew back in a hormone-independent state. It is possible that the addition of chemotherapy at the time of initial androgen ablation will improve cell kill by potentiating apoptosis, thereby killing cells that might otherwise have mutated to the androgen-independent state if allowed to continue to cycle and grow. The rationale behind Radiation Therapy Oncology Group (RTOG) P 0014 is to demonstrate in a randomized phase 3 trial that giving patients chemotherapy at the beginning of androgen blockade may improve patient survival. PMID- 14747048 TI - Novel approaches to treat asymptomatic, hormone-naive patients with rising prostate-specific antigen after primary treatment for prostate cancer. AB - Biochemical-only recurrent prostate cancer presents the ideal setting for assessing novel agents or approaches for prostate cancer treatment. There is no clear evidence that delay in initiation of more definitive androgen-deprivation therapy is harmful, and a simple blood test--the prostate-specific antigen (PSA) level--is readily available to screen for potential antineoplastic activity. Current novel approaches include vaccines, cyclooxygenase-2 (COX-2) inhibitors, selective apoptotic antineoplastic drugs, endothelin-A receptor antagonists, chemotherapy, vitamin D, and peroxisome proliferator-activated receptor-gamma agonists. In this screening process, certain therapies have emerged as delaying PSA progression or decelerating PSA velocity. These therapies, such as the COX-2 inhibitors, will need to proceed to phase 3 trials to answer the more important question of whether this change in PSA dynamics translates into improved survival. Patients enrolling in these trials need to be clearly informed of the limited expectations of these novel exploratory approaches. PMID- 14747049 TI - Use of nomograms for predicting survival in patients with castrate prostate cancer. AB - Given the heterogeneity of prognoses in patients with castrate metastatic prostate cancer, the ability to accurately predict survival is vital for optimal patient counseling, selection of treatments, clinical trial design, and interpretation of clinical data. Over the past 20 years, several prognostic models have been developed in an attempt to refine the clinician's predictive ability. Early models were based on patients with more advanced disease. They included variables that are no longer regularly encountered today and involved cumbersome calculations that were not practical for everyday use in the clinic. Recently, 2 point-based nomograms have been developed based on pretreatment variables measured on a routine basis. These models provide a user-friendly format in which to make sophisticated predictions of survival. These models have improved our ability to predict the outcomes of patients with castrate metastatic disease. However, further work to identify novel prognostic markers to improve the accuracy of these predictions is needed. PMID- 14747050 TI - Prostate-specific antigen doubling time and survival in patients with advanced metastatic prostate cancer. AB - The relation between tumor kinetics and disease progression in patients with hormone-refractory prostate cancer (HRPC) has not been well described. Biochemical recurrence of prostate cancer is characterized by detectable prostate specific antigen (PSA) levels after treatment and occurs in approximately 30% of patients after therapy for apparent localized disease. An increase in PSA almost always occurs before clinical evidence of disease. The ability to identify early biochemical failure in patients to assess disease aggressiveness and guide changes in treatment needs to be examined. We examined serial PSA data from 249 patients with metastatic disease to assess PSA doubling time (PSADT) in hormone naive prostate cancer (HNPC) and HRPC states. In a subset of patients, the relation of PSADT to Gleason score and survival was studied. PSADT decreased from 37.5 +/- 4.5 weeks to 15.6 +/- 1.6 weeks (mean +/- SEM) in patients with HNPC versus HRPC. In this small study, PSADT did not correlate with Gleason score, survival from start of hormonal treatment, length of time receiving hormone therapy, or survival in the HRPC state. The decrease in PSADT with disease state may help provide insight into understanding the biology of late-stage disease. PMID- 14747051 TI - The patient with hormone-refractory prostate cancer: determining who, when, and how to treat. AB - Hormone-refractory prostate cancer (HRPC) encompasses a wide spectrum of patients, including patients with prostate-specific antigen (PSA)--only disease, those with increasing PSA levels yet stable metastatic disease, and those with increasing PSA levels and objective evidence of progressive metastases. Unfortunately, with the historical lack of effective therapy in this population, the oncologist is faced with few data with which to make difficult clinical decisions. Although our understanding of the biology of androgen independence continues to improve, and our fund of potential therapeutic agents has widened, multiple trial-specific and patient-specific obstacles have contributed to the difficulty in demonstrating clear benefit to therapy. Herein, we will review the biology of androgen-independent prostate cancer, the historical impediments to clinical trials in this population, and the reasons to treat, or not to treat, the patient with HRPC. PMID- 14747052 TI - Phase 3 randomized trial evaluating second-line hormonal therapy versus docetaxel estramustine combination chemotherapy on progression-free survival in asymptomatic patients with a rising prostate-specific antigen level after hormonal therapy for prostate cancer: an Eastern Cooperative Oncology Group (E1899), Intergroup/Clinical Trials Support Unit study. AB - Despite improvements in early detection of prostate cancer when it is clinically localized--and therefore most amenable to curative local therapies-- approximately 33% of men with early prostate cancer will develop biochemical failure with a rising prostate-specific antigen (PSA) during follow-up. The early use of androgen suppression in this group of men has changed the clinical picture of androgen-independent disease. Now, there are men on androgen suppression who will develop biochemical failure and remain free of clinical or radiographic evidence of metastatic disease for a period of time. There is no standard approach to this group of men, who have a rising PSA on androgen deprivation, because there are little or no data about strategies that may improve survival, delay time to progression, or improve quality of life. Therefore, current management of this population remains controversial. In an effort to determine which of these 2 approaches--second-line hormone therapy or chemotherapy--is optimal in delaying the time to progression, the Eastern Cooperative Oncology Group (ECOG) developed protocol E1899. Although both approaches offer the potential for response (reduction in PSA and prolonging the time to clinical progression), they have very different toxicity profiles, making impact on quality of life another important end point. This randomized trial evaluating second-line hormonal therapy using ketoconazole and hydrocortisone versus docetaxel and estramustine combination chemotherapy on progression-free survival in asymptomatic men with a rising PSA on androgen ablation for prostate cancer is the subject of this article. PMID- 14747053 TI - Novel approaches to treating the asymptomatic hormone-refractory prostate cancer patient. AB - For >50 years, the standard treatment for advanced prostate cancer has been hormonal therapy. However, all such treated patients eventually develop disease refractory to androgen suppression as manifested by increasing prostate-specific antigen (PSA) levels, progressive disease on radiographic imaging, and ultimately, symptomatic deterioration. Historical perceptions that treatment of hormone-refractory prostate cancer was a largely futile venture have faded over the past decade with the advances in new therapeutic strategies. With the use of PSA values to follow the progress of patients after definitive therapy, physicians are seeing more patients who have failed hormonal therapy yet have no symptoms from their disease. There are standard therapies available for patients who require palliation for symptoms, but there is no consensus on treatment for asymptomatic patients. To date, there has been no definitive increase in survival with any therapy in this group of patients. In addition, several novel drugs have advanced through preclinical testing into early clinical trials. It is these drugs--alone or in combination--that are designed to target strategic tumor pathways in these patients. This article will review a selection of agents that may be potentially useful in this population. PMID- 14747054 TI - The language of surgical pathology--a precis for the head and neck surgeon. PMID- 14747055 TI - Database on monoclonal antibodies to cytokeratins. AB - Cytokeratins (CK) are being extensively used as diagnostic markers for various malignancies and other diseases, including human oral precancer and cancer, due to their tissue specific expression. CK are epithelia specific intermediate filament (IF) proteins, which are expressed in a differentiation dependent and tissue specific manner. There are about 30 polypeptides of CK expressed by different human epithelia. Each type of epithelium expresses about 4-6 polypeptides. CK polypeptides share many common epitopes, due to which the antibodies developed against CK tend to cross react. Therefore, a large number of monoclonal and polyclonal antibodies have been developed to distinguish among these proteins. Many of these antibodies are not only monospecific but are also epitope specific. These antibodies are being used in pathology laboratories for routine diagnosis using immunohistochemistry. A number of fixatives are used for fixation of tissue sections prior to the use of these antibodies. Sometimes, this leads in epitope masking. Hence, it becomes necessary to use a battery of monoclonal antibodies (MAb) for accurate diagnosis. Apart from the use of these antibodies in diagnostics, they are also being used in basic research for the study of CK function and their interactions with associated proteins and membrane proteins. In the present communication an effort has been made to make a comprehensive list of MAb to CK giving information like cross-reactivity, epitope specificity, various fixatives used, etc. along with the source of the antibodies, which will provide useful information to the users. PMID- 14747056 TI - Reduced expression of hyaluronan is a strong indicator of poor survival in oral squamous cell carcinoma. AB - Several malignant tumours accumulate hyaluronan (HA), a matrix component suggested to promote cancer cell growth and migration. The expression and prognostic value of HA was analysed in a cohort of 151 oral squamous cell carcinoma (SCC) patients with adequate archival tumour material and follow-up data. The tumour samples were stained using a biotinylated HA-specific probe. Normal squamous epithelium showed a strong and homogeneously distributed staining for HA. The most superficial layers were HA-negative. In moderate (n=11) and high grade (n=16) dysplasias an irregular HA staining was observed around invasive cancer. Malignant transformation in oral squamous cell epithelium changed the staining toward irregular with focal reduction of HA. The well (n=92) or moderately differentiated (n=47) carcinomas had a strong HA staining intensity. In poorly differentiated tumours (n=12) the HA staining was weaker and mainly intracellular. The stromal tissue showed usually moderate (n=69) or strong (n=67) HA staining intensity with no statistically significant correlation with the degree of tumour differentiation. At the end of the follow-up (median 52 months) 66 (43%) patients had died because of an oral SCC. A significant difference in overall survival (OS) and disease free survival (DFS) (P=0.0002 and 0.0020, respectively) was noticed between the patients with the different epithelial staining patterns for HA. The reduction of HA staining was associated with poor survival. In Cox's multivariate analysis HA staining was a significant independent predictor of OS (P=0.011) and DFS (P=0.013). These results suggest that HA is a prognostic marker in oral squamous cell carcinoma. PMID- 14747057 TI - A systematic review of test performance in screening for oral cancer and precancer. AB - Nine databases were searched for studies on test performance in screening for oral cancer and precancer in primary care. Of 481 papers, full texts of 47 were scrutinised by two reviewers. Only prospective investigations of population screening involving examination of the oral mucosa, with gold standard verification (examination by an expert of subjects screened positive and at least a proportion of negatives), were selected. Seven papers describing eight studies were finally included (kappa=0.83). The weighted pooled value of sensitivity, from random effects meta-analysis, was 0.848 (95% CI 0.730, 0.919). The corresponding value for specificity was 0.965 (95% CI 0.930, 0.982). Main sources of clinical heterogeneity occurred between large house-to-house case finding programmes from SE Asia, utilising primary health workers, and smaller studies from England and Japan. Meta-analysis regression showed no difference (P=0.99) in the generally high level of discriminatory ability of the test between these two groups. PMID- 14747058 TI - Midkine induced growth of ameloblastoma through MAPK and Akt pathways. AB - Midkine (MK) is expressed during tooth development and, since ameloblastoma is thought to be arisen from the epithelium of the odontogenic apparatus or its remnant tissues, the effect of MK in ameloblastoma cell growth should be examined. The expression and function of MK were examined using 37 ameloblastoma tissues and AM-1 cells, an HPV-16DNA transfected ameloblastoma cell line. We found that MK was immunohistochemically expressed in 70% of ameloblastoma cases and AM-1 cells. By stimulation with 100 ng/ml MK, the growth of AM-1 cells was accelerated two fold by the 9th day. MK could induce phosphorylation of p44/42 MAPK (Thr202/Tyr204) and Akt (Ser473 and Thr308), and by pretreatment of PD98059, MEK1 inhibitor, or LY294002, PI3K inhibitor, MK-stimulated-phosphorylation of MAPK and Akt and MK-stimulated growth of AM-1 cells were inhibited. These results suggested that MK induced growth of ameloblastoma is through the MAPK and Akt pathways. PMID- 14747059 TI - Adhesion molecule CD44 expression in non-tumour epithelium adjacent to tongue cancer. AB - An immunohistochemical study was performed of changes in the expression of adhesion molecule CD44 in 32 adjacent non-tumour epithelia (ANTE) to lingual carcinomas and in the tumour tissue, using anti-CD44 monoclonal antibody DF1485. The aim was to evaluate the importance of these changes as an early event in lingual carcinogenesis. The ANTE was histologically normal in 22 cases (68.8%) and presented epithelial dysplasia in 10 (32.2%). Three cases of normal and dysplastic ANTE, respectively, were CD44- (9.4%). Negative CD44 expression was significantly more frequent in tumours with lower percentages of CD44+ cancer cells, both in normal (P=0.014) and dysplastic (P=0.033) ANTE. The expression in normal ANTE was also significantly associated with clinical stage (P=0.040) and presence of extracapsular nodal spread (P=0.013). Therefore, loss of CD44 expression in ANTE can be considered an early event in lingual carcinogenesis and a marker of major alterations of CD44 expression in the derived tumour tissue. PMID- 14747060 TI - Interaction between the immune system and tongue squamous cell carcinoma induced by 4-nitroquinoline N-oxide in mice. AB - Squamous cell carcinoma of the oral cavity (SCC) accounts for 3% of cancers in the western world and 40% of cancers in India. The overall 5-year survival rate is only 50%. Most of the lesions appear intra-orally on the tongue. Results from a previous study demonstrated a significant increase in T and B-lymphocytes under the transformed epithelium when examining human lesions of hyperkeratosis, dysplasia and carcinoma of the tongue. In order to investigate the interaction between the host immunity and SCC, carcinogen induced SCC of the tongue was studied in mice. The water-soluble carcinogen, 4 nitroquinoline N-oxide (4NQO), was applied to BALB/c mice tongues and produced tongue SCC after a long incubation period of several months. Immunologic properties were examined systemically in the spleens and locally, at the tumor site. Examination of spleen lymphocytes from 4NQO induced mice revealed enlargement of the spleens and a significant decrease in the CD3, CD4, CD8 and CD19 cells. In the tongues, expression of TGF-beta, TNF-alpha, GM-CSF, and IL-1 beta mRNA were detected. TNF alpha protein was detected in the affected tongues using immunoassays. mRNA expression of TNF-alpha was detected in the cancerous epithelium when extracted from the connective tissue. CD11b and CD3 cells were detected in the connective tissue under the developing carcinoma. CD11b positive cells were more prominent. The infiltrate was very scattered and not prominent as the infiltrate in the human tongue tissues. These results indicate that the growing tumor affected the immune response around the tumor and systemically. Most of the cytokines, which appeared in the affected tongues, originated from the tumor surroundings, but TNF alpha was found also in the tumor. The interaction between the tumor and immune response components is important for diagnosis and treatment purposes. PMID- 14747061 TI - Genomic instability in oral squamous cell carcinoma: relationship to betel-quid chewing. AB - Genomic instability in repeated DNA sequences is exhibited by a variety of cancer types, including oral squamous cell carcinoma. Exposure to carcinogenic compounds may further increase the instability. We have used Inter-Simple Sequence Repeat (Inter-SSR) PCR methodology to detect genetic alterations in 37 oral cancer patients who had chewed betel-quid. Thirty-eight percent of DNA from tumors had genomic alterations in the sequences flanked by (CA)(8) and (GT)(8) repeats. Patients with tumor DNAs harboring genomic alterations had a two-fold higher consumption of betel-quid than patients without alterations in tumor DNA. Matched normal and tumor DNAs were also screened for microsatellite instability where four patients (10.8%) showed alterations in at least one microsatellite marker but there was no relationship between this phenotype and betel-quid chewing. These data indicate that exposure to carcinogens present in the betel-quid may contribute to genomic instability detected by inter-SSR PCR in a subset of oral tumors. PMID- 14747062 TI - An analysis of risk factors for oral cancer in young people: a case-control study. AB - The incidence of oral cancer amongst young adults is increasing in many European and high incidence countries. The aim of this study was to evaluate the major risk factors for oral cancer in young adults using a case-control design. A sample of 116 patients aged 45 years and younger, diagnosed with squamous cell carcinoma of the oral cavity between 1990 and 1997 from the south east of England were included. Two-hundred and seven controls who had never had cancer, matched for age, sex and area of residence, were recruited. The self-completed questionnaire contained items about exposure to the following risk factors: tobacco products, cannabis, alcohol and diet. Conditional logistic analyses were conducted adjusting for social class, ethnicity, tobacco and alcohol habits. All tests for statistical significance were two-sided. The majority of oral cancer patients reported exposure to the major risk factors of tobacco and alcohol even at this younger age. The estimated risks associated with tobacco or alcohol were low (OR range: 0.6-2.5) among both males and females. Only smoking for 21 years or more produced significantly elevated odds ratios (OR=2.1; 95% CI: 1.1-4.0). Exposure associated with other major risk factors did not produce significant risks in this sample. Long term consumption of fresh fruits and vegetables in the diet appeared to be protective for both males and females. The results suggest that although this younger sample exhibit similar behavioural risk factors to older oral cancer patients, the low odds produced in addition to the relatively short duration of exposure, suggest that factors other than tobacco and alcohol may be implicated in the development of oral cancer in these younger patients. PMID- 14747063 TI - Quantification of telomerase activity of regions unstained with iodine solution that surround oral squamous cell carcinoma. AB - The aim of this study was to analyze the iodine-unstained region expanding around oral squamous cell carcinoma (SCC) by quantification of telomerase activity. The epithelial dysplasia often observed around SCC is considered to cause local recurrence or a second primary cancer. However these areas are hard to distinguish from normal mucosa. To clear the border of the expanding epithelial dysplasia around SCC, we stained with 3% iodine solution, and then decided the surgical margin. We measured quantification of telomerase activity in tumor, in epithelial dysplasia, and also in normal epithelium. Thirty-three primary cases of oral SCC which have iodine-unstained region around lesions were investigated. Fluorescense-based TRAP was applied to obtain quantification of telomerase activity. We obtained the following results: histological examination confirmed that every patient's unstained region consisted of various degrees of epithelial dysplasia. The quantified telomerase activities for squamous cell carcinoma, epithelial dysplasia and normal epithelium were 53.9, 39.6 and 2.7 U/microgP, respectively, and there was a significant difference between carcinoma and normal areas, and between dysplasia and normal epithelium. Therefore, these findings suggest that the areas of epithelial dysplasia unstained by iodine consist of cells that are nearly cancerous and excessively proliferative, and that epithelial dysplasia around SCC should be resected together with the tumor. Vital staining with iodine is useful for identifying epithelial dysplasia around SCC. PMID- 14747064 TI - Expression of vascular endothelial growth factor and prognosis of oral squamous cell carcinoma. AB - The correlation between expression of vascular endothelial growth factor (VEGF) and prognosis for oral squamous cell carcinoma was investigated. Tissue samples of oral squamous cell carcinoma were obtained from 63 patients. Of these patients, 11 had stage I, 17 had stage II, 9 had stage III, and 26 had stage IV tumours. Immunohistochemical expression of VEGF was quantitatively determined by computer-assisted image analysis. The value of VEGF expression was significantly higher for the patients with poor prognosis than for those with good prognosis (P=0.0423). Regarding regional lymph node metastasis, VEGF showed no significant difference between metastasis positive and negative patients. Expression of VEGF may thus be a prognostic marker for oral squamous cell carcinoma. PMID- 14747065 TI - Immunohistochemical expression of retinoblastoma pathway proteins in normal salivary glands and in salivary gland tumours. AB - The expression of G1-phase cell-cycle regulators is commonly deregulated in human malignancies. In the present study, we investigate components of the retinoblastoma (RB) pathway in normal salivary glands (NSG) and in salivary gland tumours (SGT). Samples of NSG, pleomorphic adenoma (PA), adenoid cystic carcinoma (ACC), mucoepidermoid carcinoma (MEC), epithelial-myoepithelial carcinoma (EMC), malignant myoepithelioma (MEM), carcinoma ex pleomorphic adenoma (CEPA), and polymorphous, low-grade adenocarcinoma (PLGA) were examined immunohistochemically using antibodies to cyclin D1, cyclin-dependent kinase 4 (CDK-4), retinoblastoma protein (pRb), CDK inhibitor p16 and transcription factor E2F-1. In normal salivary glands, cyclin D1 and cdk-4 were not expressed in any case while p16 was positively expressed. pRb was abundant and E2F-1 moderately expressed. In tumors, cdk-4 was overexpressed in half of the cases. Most tumour cases showed decreased pRb immunoexpression compared to normal salivary glands. In contrast, expression of p16 and E2F-1 increased. pRb expression was absent in three cases of PA, two of EMC and one of CEPA. One case of MEM and one of PLGA showed no E2F-1 expression. Statistical analyses revealed positive correlations between cyclin D1 and cdk-4, cyclin D1 and E2F-1, cdk-4 and E2F-1, and p16 and E2F-1. The benign and malignant tumours expressed retinoblastoma pathway proteins differently form the normal salivary gland. Our findings suggest that, pRb pathway deregulation in salivary gland neoplasms is unrelated to their biological behaviour. PMID- 14747066 TI - Instability of background fat intensity suppression using fat-saturated (FS) MR imaging techniques according to region and reconstruction procedure in patients with oral cancer. AB - The purpose of the present study is to evaluate the background fat intensity suppression instability of each area in the head and neck region, and in the post reconstruction with metal plate and myocutaneous flap, of patients with oral cancer using fat-saturated (FS) images. STIR and FS T2-weighted images at pre- and post-surgery in 59 patients with oral cancer were scored for uniformity of fat suppression and tissue conspicuity in each region of the head and neck. The scores of FS on uniformity of fat suppression pre-operatively were worse than those of STIR in the mandibular level, but not lesion and tissue conspicuity. However, the deterioration both of scores between pre- and post-surgery using FS was worse than that using STIR using metal plate and/or myocutaneous flap. At diagnosis, we should recognize on MR images using FS that instability of the status of fat suppression might be brought about by respective area and reconstruction with metal plate and myocutaneous flap of patients with oral cancer. PMID- 14747067 TI - Dendritic cell sarcoma of the oral cavity. AB - Sarcomas derived from immune accessory cells are uncommon malignancies, most of them occurring in lymph nodes while extra nodal sites are very rarely affected. Based on the immune profile, the cells that give origin to these neoplasms are currently divided in: follicular dendritic cell (FDC), interdigitating dendritic cell (IDC), indeterminate cell and Langerhans cell. A case of a dendritic cell sarcoma arising in the alveolar ridge mucosa in a 50-year-old female is reported here. The lesion presented as a nodular mass without defined borders and covered by reddish mucosa. Histologically, the tumour was composed of spindle-shaped cells with large nuclei and abundant cytoplasm, arranged in variable patterns as storiform and whorled and revealing interspersed lymphocytes. No capsule could be seen and the neoplasm extended up to the lining epithelium. Immunohistochemically, the spindle cells were positive for vimentin, S-100 protein, CD1a, factor XIIIa and focally to smooth-muscle actin, but were negative for CD21, CD35, CD23 and caldesmon--all markers of follicular dendritic cells. In conclusion, the present case has morphologic pattern of dendritic cell sarcoma and the immunophenotype is compatible with IDC cells or with intermediate dendritic cells and demonstrates the overlap of features among these entities. PMID- 14747068 TI - Hyalinizing clear cell carcinoma of salivary gland: an aggressive variant. AB - Although hyalinizing clear cell carcinoma (HCCC) had been previously illustrated by several authors, it was not until 1994 that this tumour was characterized by Milchgrub et al. [Am J Surg Pathol (1994),18,74] and separated from the heterogeneous group of clear cell carcinomas described in the literature. HCCC is a distinctive infiltrative low-grade, monomorphic, glycogen-rich clear cell carcinoma with prominent stromal hyalinization occurring most often in the minor salivary glands of adult women. A case of hyalinizing clear cell carcinoma arising in the tongue of an adult female is described with special reference to the presence of minor foci of mitotic activity, necrosis and anaplasia in this otherwise typical low-grade carcinoma. Widespread metastases and death within a year of initial presentation in this case suggests that there may be a subset of this indolent tumour in which these features are associated with a poor prognosis. PMID- 14747069 TI - Composite lymphoma: angiocentric T-cell lymphoma (CD8+ cytotoxic/suppressor T cell) and diffuse large B-cell lymphoma associated with EBV, and presenting clinically as a midfacial necrotizing lesion. AB - A composite lymphoma is defined as the simultaneous occurrence of two histologically different types of lymphomas situated in one anatomical location. Reports of composite B- and T-cell lymphomas, especially in the head and neck region, are rare. We describe a 76-year-old Taiwanese aboriginal female patient clinically presenting with a midfacial necrotizing lesion (MNL). Microscopic examination of the incisional biopsy specimen revealed extensive surface necrosis with infiltrates of inflammatory cells. Beneath the necrotic surface, there appeared to be two distinct populations of pleomorphic lymphoid cells exhibiting the characteristic features of the angiocentric distribution of the tumor cells and evidence of angiodestruction. Immunohistochemical staining revealed that these atypical lymphoid cells were positive for LCA, CD45, CD5, CD20, CD3 epsilon, CD8, bcl-2 and bcl-6 and negative for CD56, CD4, CD68, keratin, S-100, kappa and lambda. Furthermore, these atypical lymphoid cells also expressed EBV encoded nuclear RNAs (EBERs) following in situ hybridization. Therefore, this was a case of composite lymphoma: angiocentric T-cell lymphoma (ATCL) (CD8+ cytotoxic/suppressor T-cell) and diffuse large B-cell lymphoma (DLBL) associated with the Epstein-Barr virus (EBV) and presenting clinically as MNL. PMID- 14747070 TI - Investigations of the EPR parameters for Sm3+ ions in KY3F10 and LiYF4 crystals. AB - In this paper, we calculate the EPR parameters (g factors g parallel, g perpendicular and hyperfine structure constants A parallel, A perpendicular) of rare earth ion Sm3+ in fluoride crystals KY3F10 and LiYF4 from the perturbation formulas of EPR parameters for a 4f5 ion in tetragonal symmetry. In these formulas, the crystal-field J-mixing of the first and second excited-state multiplets 6H(7/2) and 6H(9/2) into the ground state multiplet 6H(5/2), the mixtures among the states with the same J value via spin-orbit coupling interaction and the interactions between the ground Kramers doublet Gammagamma and the same irreducible representation as Gammagamma in other 11 Kramers doublets Gammax within 6HJ (J=5/2, 7/2, 9/2) states via crystal-field and orbital angular momentum (or hyperfine structure) are considered. The calculated results (which are in agreement with the observed values) are discussed. PMID- 14747071 TI - Frequency measuring system using mirror gap stabilized Fabry-Perot interferometer. AB - The mirror gap of a Fabry-Perot interferometer was stabilized with two laser diodes; one locked to the line frequency 385,243,555.14445 MHz of the F=3<--1 in 5D(3/2)<--5S(1/2) (87Rb) and the other to the 385,284,566.3663 MHz of the F=4<--2 in 5D(5/2)<--5S(1/2) (87Rb) [Opt. Commun. 102 (1993) 432]. The length of the mirror gap was adjusted to generate the zero-cross points simultaneously at both of the two line positions. The fringe signals obtained from an interferometer thus stabilized can be used as frequency markers having accuracies of the order of 10(10) depending on the finesse of the interferometer used. Based on measurement using Cs D1 hyperfine lines reported by Udem et al. [Phys. Rev. Lett. 82 (1999) 3568], the uncertainty of markers in the region 50 THz apart from the reference lines is +/- 5 MHz. PMID- 14747072 TI - Characterization of zinc hydroxynitrates by diffuse reflectance infrared spectroscopy--structural modifications during thermal treatment. AB - Four zinc hydroxynitrates are known, but only the Infrared spectrum of Zn5(OH)8(NO3)2.2H2O is reported in the literature. Here, we report the IR spectra of the other three zinc hydroxynitrates, namely Zn(OH)(NO3).H2O, Zn3(OH)4(NO3)2 and Zn5(OH)8(NO3)2. The positions of the bands of the nitrate groups between 1000 and 1600 cm(-1) are discussed in relation to the structure of these compounds. The modifications of these IR spectra with temperature are consistent with previous studies on the thermal decomposition of these compounds. PMID- 14747073 TI - Infrared spectroscopic study on the thermal decomposition of external and internal gelation products of simulated mixed oxide nuclear fuel. AB - The thermal decomposition of urania-ceria gel corresponding to the composition U(0.7)Ce(0.3)O(2+x) obtained through external and internal gelation routes were studied using infrared spectroscopy (IR). In the case of externally gelated compound, the gel decomposes with the release of H2O and NH3 below 500 degrees C. A part of the NH3 released is entrapped in the solid and above 500 degrees C self reduction occurs in which U(VI) in the gel is reduced to U3O8. The decomposition products were identified to be U3O8 and CeO2. In the case of internally gelated compound, decomposition similar to the one for externally gelated compound occurred below 500 degrees C. Above 500 degrees C the carbon present in the gel reduced U(VI) to UO2 which formed solid solution with CeO2 around 650 degrees C. PMID- 14747074 TI - Vibrational spectra and ab initio analysis of tert-butyl, trimethylsilyl, and trimethylgermyl derivatives of 3,3-dimethyl cyclopropene V. 3,3-dimethyl-1 (trimethylgermyl)cyclopropene. AB - 3,3-dimethyl-1-(trimethylgermyl)cyclopropene (I) was synthesised using a standard procedure. The IR and Raman spectra of I in the liquid phase were measured. The molecular geometry of I was optimised completely at the HF/6-31G* level. The HF/6 31G*//HF/6-31G* force field was calculated and scaled using the set of scale factors transferred from those determined previously for scaling the theoretical force fields of 3,3-dimethylbutene-1 and 1-methyl-, 1,2-dimethyl-, and 3,3 dimethylcyclopropene. The assignments of the observed vibrational bands were performed using the theoretical frequencies calculated from the scaled HF/6 31G*//HF/6-31G* force field and the ab initio values of the IR intensities, Raman cross-sections and depolarisation ratios. The theoretical spectra are given. The completely optimised structural parameters of I and its vibrational frequencies are compared with corresponding data of related molecules. PMID- 14747075 TI - Raman properties of chlorophyll d, the major pigment of Acaryochloris marina: studies using both Raman spectroscopy and density functional theory. AB - The Raman spectroscopy of purified chlorophyll (Chl) d extracted from Acaryochloris marina has been measured over the wide region of 250-3200 cm(-1) at 77 K following excitation of its Soret band at 488 nm and analyzed with the aid of hybrid density-functional vibrational analyses. A Raman peak specific to Chl d, which arises from the formyl group 3(1) C=O stretching, was clearly observed at 1659 cm(-1) with medium intensity. Peaks due to other C=O stretching vibrations of the 13(1) keto-, 13(3) ester- and 17(3) groups were also observed. Four very strong peaks were observed in the range of 1000-1600 cm(-1), assigned to the CC stretching and mixtures of the CH3 bend and CN stretching. CCC and NCC bending contribute to medium intensity peaks at 986 and 915 cm(-1). Out-of-plane CH bending at Chl d methine sites 10, 5 and 20 contribute to observed peaks at 885, 864 and 853 cm(-1), respectively. A few modes involving the MgN stretching and MgNC bending motions were observed in the very low frequency range. Density functional theory (DFT) calculations have been used to make assignments on the observed Raman spectrum and the DFT results have been found to be in good agreement with the experimental results. PMID- 14747076 TI - Assignment of charge transfer absorption band in optical absorption spectra of the adsorbate-silver colloid system. AB - In this paper we reported the UV-visible-NIR optical absorption properties of silver colloid, employed as a high efficient substrate in surface-enhanced Raman spectra (SERS), under various conditions. Experimental results revealed that the new absorption band, usually appearing in the longer wavelength region due to the addition of molecules, was related to the direct adsorption of molecules on colloidal silver surface. When the adsorption occurred, this new band would appear. Once the molecules were desorbed from silver surface, the new band could not be observed. Some evidences inferred that the new absorption band was associated with the effect of charge-transfer transition between adsorbates and colloidal silver particles, while not with the effect of the surface plasma resonance due to the silver particles aggregation which was usually attributed to in previous research work. PMID- 14747077 TI - Vibration assignment of carbon-sulfur bond in 2-thione-1,3-dithiole-4,5 dithiolate derivatives. AB - The higher frequency peak near 1050 cm(-1) of the doublet in the infrared and Raman spectra of 2-thione-1,3-dithiole-4,5-dithiolate (DMIT) derivatives corresponds mainly to the stretching vibration of carbon-sulfur double bond in the terminal S=CS2 fragments of DMIT skeletons while the lower one corresponds mainly to the Fermi resonance peak between overtones of symmetric stretching vibration of two carbon-sulfur single bond in the S=CS2 fragment of DMIT skeletons and the higher frequency. On the other hand, the higher frequency near 500 cm(-1) corresponds mainly to the symmetric stretching vibration of two carbon sulfur single bond in the S=CS2 fragments of DMIT skeletons while the lower one corresponds mainly to the symmetric stretching vibration of the four carbon sulfur single bond in the S2C=CS2 fragments of DMIT skeletons. PMID- 14747078 TI - Absorption and steady state fluorescence study of interaction between eosin and bovine serum albumin. AB - The interaction between eosin and bovine serum albumin in buffer solution, at pH 7.4 has been studied by means of absorption and emission spectroscopy. Applying the Scatchard model to the absorbance data a non-linear plot was obtained, reflecting a complex process. In the fluorescence spectra, two distinct effects were observed. Upon increasing the protein-dye ratio to about 0.60, the intensity of eosin emission band (544 nm) decreases to approximately 30% of its initial value. During this quenching, a small red shift is noticed. The data were rationalized in terms of two classes of binding sites. At higher protein concentrations, a new band localized at 556 nm appears, which could be assigned to a new fluorescent species. This second process corresponds to a 1:1 binding. PMID- 14747079 TI - Temperature-dependent Raman spectra of collagen and DNA. AB - Raman spectra of collagen and DNA were discussed at different temperatures. The temperature-dependence of Raman intensity was obtained in the region from -150 to 200 degrees C. Four denaturation points at 0, 40, 68 and 90 degrees C of collagen and two peaks at 38 and 82 degrees C for DNA were obtained. The wavenumbers of many vibrational modes were found to increase for lower temperature, but the peak at 1302 cm(-1) of collagen and the peak at 1101 cm(-1) of DNA showed the opposite trend. In all of the vibrational modes of DNA, the bases showed the most sensitive to different temperatures and there is a pronounced shift of bands at 70 degrees C, the starting point of denaturation. PMID- 14747080 TI - The detection of drugs of abuse in fingerprints using Raman spectroscopy I: latent fingerprints. AB - This paper describes the application of Raman spectroscopy to the detection of exogenous substances in latent fingerprints. The scenario considered was that of an individual handling a substance and subsequently depositing a contaminated fingerprint. Five drugs of abuse (codeine phosphate, cocaine hydrochloride, amphetamine sulphate, barbital and nitrazepam) and five non-controlled substances of similar appearance, which may be used in the adulteration of drugs of abuse (caffeine, aspirin, paracetamol, starch and talc), were studied in both sweat rich and sebum-rich latent fingerprints. The substances studied could be clearly distinguished using their Raman spectra and were all successfully detected in latent fingerprints. Photobleaching was necessary to reduce the fluorescence background in the spectra of some substances. Raman spectra obtained from the substances in sweat-rich latent fingerprints were of a similar quality to spectra that obtained from the substances under normal sampling conditions. Interfering Raman bands arising from latent fingerprint material were present in the spectra obtained from the substances in sebum-rich fingerprints. These bands did not prevent identification of the substances and could be successfully removed by spectral subtraction. The most difficult aspect of the detection of these substances in latent fingerprints was visually locating the substance in the fingerprint in order to obtain a Raman spectrum. PMID- 14747081 TI - Spectrophotometric study of the system Hg(II)-thymol blue-H2O and its evidence through electrochemical means. AB - The detailed analysis of the experimental spectrophotometric data obtained from solutions containing the acid-base indicator thymol blue (TB) and mercury(II) (Hg(II)) coupled with data processing by means of the SQUAD program, a chemical model was determined that includes the formation of complexes indicator-metal ion (HgTB and HgOTB), dimer species (H3TB2 and H4TB2) and monomer species (HTB and TB). The values of the overall formation constants (log beta) were calculated for the chemical equilibria involved: TB+Hg<-->HgTB log beta=16.047 +/- 0.043, TB+Hg+H2O<-->HgOHTB+H log beta=7.659 +/- 0.049, 2TB+4H<-->H4TB2 log beta=31.398 +/- 0.083, 2TB+3H<-->H3TB2 log beta=29.953 +/- 0.084 and H+TB<-->HTB-log beta=8.900. To compliment the present research, the values of the absorptivity coefficients are included for all the species involved, within a wide range of wavelengths (250-700 nm). The latter were used subsequently to carry simulations of the absorption spectra at various pH values, thus corroborating that the chemical model proposed is fully capable to describe the experimental information. Voltammetric study performed evidenced the formation of a complex with a 1:1 stoichiometry Hg(II):TB. PMID- 14747082 TI - Simultaneous photometric determination of albumin and total protein in animal blood plasma employing a multicommutated flow system to carried out on line dilution and reagents solutions handling. AB - An automatic flow procedure for the simultaneous determination of albumin and total protein in blood plasma samples is proposed. The flow network comprised a set of three-way solenoid valves assembled to implement the multicommutation. The flow set up was controlled by means of a computer equipped with an electronic interface card which running a software wrote in QUICKBASIC 4.5 performed on line programmed dilution to allow the determination of both albumin and total protein in blood plasma. The photometric methods based on Bromocresol Green and Biuret reagents were selected for determination of albumin and total protein, respectively. Two LEDs based photometers coupled together the flow cells were employed as detector. After the adjustment of the operational parameters the proposed system presented the following features: an analytical throughput of 45 sample processing per hour for two analytes; relative standard deviations of 1.5 and 0.8% (n=10) for a typical sample presenting 34 g l(-1) albumin and 90 g l(-1) total protein, respectively; linear responses ranging from 0 to 15 g l(-1) albumin (r=0.998) and total protein (r=0.999); sample and reagents consumption, 140 microl serum solution, 0.015 mg VBC and 0.432 mg CuSO4 per determination, respectively. Applying the paired t-test between results obtained using the proposed system and reference methods no significant difference at 95 and 90% confidence level for albumin and total protein, respectively, were observed. PMID- 14747083 TI - In situ UV-Vis spectroelectrochemical studies to identify electrochromic sites in poly(1-naphthylamine) modified by diphenylamine. AB - Poly(1-napthylamine) (PNPA) was modified by incorporation of diphenylamine (DPA) to result copolymer film on the indium tin oxide (ITO) coated glass electrode using cyclic voltammetry. Detailed static and dynamic UV-Vis spectroelectrochemical studies were performed on the copolymer films to identify the electrochromic sites corresponding to individual DPA and NPA units. Absorbance-wavelength-potential (AWP) profiles were constructed from the dynamic spectroelectrochemical results to infer the electrochromic sites. The advantages of using AWP profile in the analysis of electrochromic sites are discussed. PMID- 14747084 TI - Fluorescence spectrum of the benzyl radical in methylcyclohexane at 4.2 K. AB - The fluorescence spectrum of the benzyl radical (BR) is characterized by participation of many totally symmetric vibrational modes with less intensity activeness. To make this point more clear, the fluorescence spectra of the benzyl radical and its deuterated compounds in methylcyclohexane (MCH) have been observed at 4.2 K and detailed vibrational analysis has been carried out. Two non totally symmetric modes such as nu6b and nu8b play a role of Herzberg-Teller type vibronic coupling together with one totally symmetric mode nu6a. There are only a few progression bands with very weak intensity. These are nu12, nu1 and nu(phi CH2) vibrational modes. PMID- 14747085 TI - On the 1,3-dipolar cycloaddition reactions of indenone with N-N-C dipoles: density functional theory calculations. AB - Density functional theory (DFT) calculations at the B3LYP/6-311G* theoretical level have been performed to study the 1,3-dipolar cycloaddition (1,3-DC) reactions between indenone (1) and different 1,3-dipoles (diazomethane and N methyl C-methoxy carbonyl nitrilimine, compounds 2 and 3, respectively). The geometrical and energetic properties were analysed for the different reactives, transition states and cycloadducts formed (compounds 4-11). The reactions proceed in the gas-phase by an asynchronous concerted mechanism, yielding different regiochemistry dependent on the 1,3-dipole chosen, although with dipole 3 some degree of synchrony was found in the formation of cycloadduct 5. The 1,3-DC between 1 and 3 was regioselective, being the cycloadduct 11 favoured against 9. The NMR chemical shift parameters (GIAO method) were also calculated for the reactives and cycloadducts. PMID- 14747086 TI - Experimental and theoretical vibrational study of 2,2,2-trifluoroethyl trifluoromethanesulfonate, CF3SO2OCH2CF3. AB - The infrared spectra of 2,2,2-trifluoroethyl trifluoromethanesulfonate (CF3SO2OCH2CF3) were obtained in the gaseous, liquid and solid states as well as the Raman spectrum of the liquid. Quantum chemistry calculations using the density functional theory were used to predict the most stable geometry and conformation of the studied molecule. Subsequently, the harmonic vibrational frequencies and force field were calculated. An assignment of the observed spectral features made after comparison with the related molecules and with the predicted frequencies was used as the basis of a scaling of the original force field in order to reproduce as well as possible the experimental frequencies. With this purpose a set of scale factors was calculated by a least square procedure, leading to a final root mean square deviation (RMSD) of 9.7 cm(-1). PMID- 14747087 TI - Vibrational frequencies and structural determinations of tetrachlorobutatriene. AB - The normal mode frequencies and corresponding vibrational assignments of tetrachlorobutatriene in D2h symmetry are examined theoretically using the Gaussian 98 set of quantum chemistry codes. All normal modes were successfully assigned to one of the six types of motion (C=C stretch, CCl2 scissors, CCl2 twist, CCl2 wag, CCl2 rock, and C=C=C bend) predicted by a group theoretical analysis. By comparing the vibrational frequencies with IR and Raman spectra available in the literature, a set of scaling factors is derived. PMID- 14747088 TI - Vibrational frequencies and structural determination of trichloroboroxine. AB - The normal mode frequencies and corresponding vibrational assignments of trichloroboroxine (B3O3Cl3) in D3h symmetry are examined theoretically using the Gaussian 98 set of quantum chemistry codes. All normal modes were successfully assigned to one of five types of motion (B-Cl stretch, B-O stretch, B-Cl bend, O B-O bend, and B(OOCl) umbrella motion) predicted by a group theoretical analysis. By comparing the vibrational frequencies with IR and Raman spectra available in the literature, a set of scaling factors is derived. Molecular orbitals and bonding are examined. PMID- 14747089 TI - Optical properties of Sm3+ doped calibo glass with addition of lead oxide. AB - Optical absorption and fluorescence spectra of Sm3+ doped calibo glasses and the effects on these on addition of lead oxide have been recorded. The concentration of lead oxide has been varied from 0 to 30 mol%. The Judd-Ofelt intensity parameters Omegalambda (lambda=2, 4, 6) for f-f transitions in Sm3+ ions trapped in these glasses are calculated and their variation with increase in proportion of lead oxide has been discussed. It is observed from the fluorescence spectra that the intensity of fluorescence reaches a maximum when 20% of PbO is present. Stimulated emission cross-section is calculated and compared with its value for other lattices. PMID- 14747090 TI - A Raman spectroscopic study of thermally treated glushinskite--the natural magnesium oxalate dihydrate. AB - Raman spectroscopy has been used to study the thermal transformations of natural magnesium oxalate dihydrate known in mineralogy as glushinskite. The data obtained by Raman spectroscopy was supplemented with that of infrared emission spectroscopy. The vibrational spectroscopic data was complimented with high resolution thermogravimetric analysis combined with evolved gas mass spectrometry. TG-MS identified two mass loss steps at 146 and 397 degrees C. In the first mass loss step water is evolved only, in the second step carbon dioxide is evolved. The combination of Raman microscopy and a thermal stage clearly identifies the changes in the molecular structure with thermal treatment. Glushinskite is the dihydrate phase in the temperature range up to the pre dehydration temperature of 146 degrees C. Above 397 degrees C, magnesium oxide is formed. Infrared emission spectroscopy shows that this mineral decomposes at around 400 degrees C. Changes in the position and intensity of the CO and CC stretching vibrations in the Raman spectra indicate the temperature range at which these phase changes occur. PMID- 14747091 TI - Preparation and analysis by UV-Vis of zinc phthalocyanine complexes. AB - We have prepared and isolated a series of zinc phthalocyanine complexes (ZnPc, ZnPcF8, biZnPc, F12biZnPc). These compounds have been analyzed by electronic spectrometry. The analysis of results demonstrates that the importance of intramolecular interactions in the binuclear species depends on the nature of the peripheral substituents. PMID- 14747092 TI - Vibrational spectrum, ab initio calculations, conformational equilibria and torsional modes of 1,3-dichloropropane. AB - Ab initio calculations are reported for three of four possible conformers of 1,3 dichloropropane. The fourth conformer, with Cs symmetry, has a predicted enthalpy difference of more than 1500 cm(-1) from the most stable conformer from each calculation regardless of the basis set used, so there is little chance of observing it. Thus, there is no evidence in the infrared or Raman spectrum of the presence of a fourth conformer. The order of stability given by the ab initio calculations is C2(GG)>C1(AG)>C2v(AA)>Cs(GG'), where A indicates the anti form for one of the CH2Cl groups and G indicates the gauche conformation for the other CH2Cl group relative to the plane of the carbon atoms. Almost every band observed can be confidently assigned to one or another of the conformers. Many observed bands proved to be of a composite nature, with several nearly coincident vibrations of different conformers contributing to the band contour. Nonetheless, a complete assignment of fundamentals is possible for the most stable C2 conformer, and 5 of the fundamentals of the C2v conformer and 13 those of the C1 conformer can be confidently assigned. PMID- 14747093 TI - Study of the interaction of trimethoprim-montmorillonite by infrared spectroscopy. AB - The interaction of trimethoprim with montmorillonite has been studied under different pH conditions and ionic strengths. Infrared spectra indicate that the interaction is favoured by cation-exchange mechanism at low pH, and by physical adsorption at high pH. This is also supported by adsorption isotherms and the desorption process which is reversible at low acidity. PMID- 14747094 TI - Theoretical studies on vibrational spectra of some halides of Group IIB elements. AB - The vibrational spectra of Group IIB elements halides MX2 and their dimers M2X4 (M=Zn(II), Cd(II) and Hg(II); X=F, Cl, Br and I) have been systematically investigated by ab initio RHF and B3LYP methods with LanL2MB, LanL2DZ and SDD basis sets. The optimized geometries, calculated vibrational frequencies are evaluated via comparison with the experimental data. The vibrational frequencies, calculated by these methods with different basis sets, are compared to each other too. The best results can be obtained by RHF/SDD method, with this method, the deviations for MX2 and Hg2X4 are <7%. Some vibrational frequencies of M2X4 that have not been experimentally reported are also predicted. PMID- 14747095 TI - Lattice vibrations of manganese oxides. Part I. Periodic structures. AB - Raman scattering (RS) and Fourier transform-infrared (FT-IR) spectroscopy have been applied to the structural characterisation of manganese dioxides (MDOs). A variety of synthetic battery-grade MDOs are investigated for comparison to the natural phases. The RS and FT-IR spectra are analysed on the basis of the local environment in the MDO structures considering the vibrations of the MnO6 octahedral building the lattices. The vibrational modes of the MnO6 units expand over 400-650 cm(-l) with additional bands in the low-wavelength region. Structural trends are deduced from the comparison of the vibrational spectra of the MDO phases investigated: birnessite, bixbyite, coronadite, groutite, hausmannite, hollandite, manganosite, pyrolusite, ramsdellite, romanechite, spinel, and todorokite. PMID- 14747096 TI - Theoretical studies of the spin Hamiltonian parameters and the local structures for M2+ (M=Co, Mn, V and Ni) ions in CsMgCl3. AB - The spin Hamiltonian parameters (zero-field splitting D, g factors g parallel, g perpendicular and hyperfine structure constants A parallel, A perpendicular) for M2+ (M=Co, Mn, V and Ni) ions in CsMgCl3 are studied by using the perturbation formulas of the spin Hamiltonian parameters for 3dn (n=7, 5, 3, 8) ions in trigonal symmetry based on the cluster approach. In these formulas, the contributions to the spin Hamiltonian parameters from the admixture of d orbitals of the central ions with the p orbitals of the ligands and from the trigonal distortion are included and the parameters related to these effects can be obtained from the optical spectra and the local structures of the studied systems. Based on the studies, it is found that the local trigonal distortion angle beta in the M2+ impurity center is unlike that betaH (approximately 51.71 degrees) in the host CsMgCl3. The spin Hamiltonian parameters for these divalent ions in CsMgCl3 are also satisfactorily explained by using the local angle beta. The validity of the results is discussed. PMID- 14747097 TI - FT Raman and FT-IR spectral studies of 3-mercapto-1,2,4-triazole. AB - The FT-IR and FT Raman spectra of 3-mercapto-1,2,4-triazole have been recorded. The observed frequencies were assigned to various modes of vibrations on the basis of normal coordinate calculations, assuming Cs point group symmetry. The potential energy distribution associated with normal modes is also reported here. The assignment of fundamental vibrations agrees well with the calculated frequencies. PMID- 14747098 TI - Investigations of EPR parameters and local lattice distortions for both Co2+ centers in La2Mg3(NO3)12.24H2O crystal. AB - The EPR g factors, g parallel and g perpendicular, of Co2+ and hyperfine structure constants (A parallel, A perpendicular) of 59Co2+ and 60Co2+ isotopes in both trigonal Mg2+ sites of La2Mg3(NO3)12.24H2O crystal are calculated from the high-order perturbation formulas of EPR parameters based on the cluster approach for 3d7 ion. It is found that to explain reasonably all these EPR parameters, the local relaxation effects (particularly, those related to the trigonal distortion angles thetai) in the vicinity of both Co2+ impurities should be considered. The local angles thetai are obtained from the calculations and the results are discussed. PMID- 14747099 TI - Electronic transitions of guanine tautomers, their stacked dimers, trimers and sodium complexes. AB - Planar and nonplanar geometries of the keto-N9H and keto-N7H tautomers of the guanine base of DNA as well as the hydrogen bonded complexes of these species with three water molecules each were optimized using the density functional theory at the B3LYP/6-31G** level. Geometries of the isolated bases were also optimized using the ab initio approach at the MP2/6-31G** level. The isolated keto-N9H and keto-N7H tautomers as well as their hydrogen bonded complexes with three water molecules each were solvated in bulk water employing the polarized continuum model (PCM) of the self-consistent reaction field theory (SCRF). Stacked dimers and trimers of both the tautomers of guanine were generated by placing the planar forms of the species at a fixed distance of 3.5 A from the neighboring one and rotating one molecule with respect to the other by 110 degrees for the keto-N9H form and 90 degrees for the keto-N7H form which corresponded to total energy minima at the B3LYP/6-31G** level. Geometry optimization for the cation of the monomer of guanine was performed at the same level of theory, and its solvation in bulk water was treated using the PCM model of the SCRF theory. The geometries of complexes of the two tautomers of guanine with a Na+ ion each were optimized at the B3LYP/6-31G** level, and the Na+ ion is predicted to bind with the keto-N9H tautomer preferentially. While the complex of the keto-N7H form of guanine with three water molecules in gas phase is slightly more stable than the corresponding complex of the keto-N9H form of guanine, the reverse is true in bulk water. Stacking interactions enhance the relative stability of the keto-N9H tautomer over that of the keto-N7H tautomer, suggesting that in bulk solutions, the former would be dominant. Electronic spectra of the isolated tautomers of guanine, those of their complexes with three water molecules each, the (keto-n9h and keto-n7h) cation of guanine, the complexes of the tautomers with a Na+ ion each, the stacked dimers and trimers of the two tautomers were calculated using configuration interaction involving single electron excitations (CIS). The relative absorption intensities of the two tautomers of guanine near 275 and 248 nm in the monomer, dimer, and trimer are predicated to be in the opposite order. Thus the absorption intensity oscillation observed using a guanine aqueous solution can be explained in terms of oscillation of relative populations of the two tautomers of the molecule. The 248 nm absorption peak would be appreciably red-shifted on formation of the cation of guanine. Binding of the Na+ ion with the two tautomers of guanine reduces intensities of their transitions appreciably and also it causes large red-shifts in the same. PMID- 14747100 TI - Preparation and study on the novel solid inclusion complex of ciprofloxacin with HP-beta-cyclodextrin. AB - The interaction of ciprofloxacin with HP-beta-cyclodextrin (HP-beta-CD) has been studied by several analytical techniques, including 1H nuclear magnetic resonance (NMR), 13C NMR, fluorescence spectra, infrared spectroscopy, thermal analyzer and scanning electron microscope. In this paper, solid inclusion complex of ciprofloxacin with HP-beta-CD was synthesized by the coprecipitation method. In addition, the characterization of the inclusion complex has been proved by fluorimetry, infrared, differential scanning calorimetry and one-dimensional (1D), 2D NMR. The experimental results confirmed the existence of 1:1 inclusion complex of ciprofloxacin with HP-beta-CD. The formation constant of complex was determined by fluorescence method and 1H NMR. Spacial configuration of complex has been proposed on two-dimensional NMR technique. PMID- 14747101 TI - Comment on: "The Renner effect in triatomic molecules with application to CH2+, MgNC and NH2" by P. Jensen, T.E. Odaka, W.P. Kraemer, T. Hirano and P.R. Bunker. (Spectrochimica Acta A58 763-794 (2002)). PMID- 14747103 TI - Thoracic surgery: is the future optimistic? PMID- 14747104 TI - Staged pulmonary and hepatic metastasectomy in colorectal cancer--is it worth it? AB - OBJECTIVES: Resection of isolated pulmonary and hepatic metastases from colorectal cancer can offer potential cure. However, little data is available to determine the results of staged hepatic and pulmonary resections in the same patient. METHODS: We retrospectively reviewed all patients who underwent staged hepatic and pulmonary metastasectomy for colorectal cancer in our institute from September 1998 to May 2002. Probability of survival was estimated by the Kaplan Meier method. RESULTS: Thirty-three metastasectomies (seven redo) were carried out in 26 patients. There were 19 male and 7 female patients with a mean age of 61 years (range 34-76 years). The mean disease-free interval for hepatic and pulmonary resection was 21.8 and 23.9 months, respectively. Sternotomy, thoracotomy and video assisted thoracoscopic approach were used in 3.03, 72.7 and 24.2% of patients, respectively. Wedge excision, lobectomy and pneumonectomy were carried out in 87.87, 9.09 and 3.03% of cases, respectively. There was one hospital death following acute respiratory failure after pneumonectomy. Mean follow-up was 23.3 months (range 2-71 months). The mean survival after last pulmonary resection was 34.7 months (SE 3.03 and 95% CI of 28.8-40.6). CONCLUSIONS: Our results support aggressive surgical management of pulmonary and hepatic metastases in colorectal cancer. PMID- 14747105 TI - Inflammatory myofibroblastic tumor of the lung. AB - OBJECTIVE: Inflammatory myofibroblastic tumor (IMT) is a rare disease that usually occurs in the lung. Recently, several reports have suggested that IMT is a true neoplasm rather than a reactive lesion. In this retrospective study, we reviewed clinicopathological characteristics and prognoses for all patients with surgically resected IMT of the lung at our institute. METHODS: From January 1985 to December 2002, nine patients had surgical intervention for IMT of the lung at the National Cancer Center Hospital, Tokyo. The resected lesions were studied histologically, immunohistochemically, and ultrastructurally. Follow-up was complete in all patients and varied from 3 months to 16 years 2 months (median, 6 years 2 months). RESULTS: These nine patients included five men and four women. They ranged in age from 25 to 66 years. Seven patients were asymptomatic. The two symptomatic patients had problems including cough, hemoptysis, and dyspnea. For all these patients, the diagnostic procedure was surgical excision. The resected tumor size ranged from 1.0 to 4.0 cm in diameter. Histologically, a variety of inflammatory and spindle cells were observed. The spindle cells corresponded ultrastructurally to myofibroblasts or fibroblasts. With the exception of one patient who had spontaneous resolution of a recurrent tumor, there was no recurrence in these patients, and all of them are in good health. CONCLUSIONS: Histopathologically, IMT is characterized by myofibroblasts that are mixed with chronic inflammatory cells, including plasma cells, lymphocytes, and histiocytes. Surgical resection, when possible, can be chosen as the treatment. Complete resection leads to excellent survival. PMID- 14747106 TI - Efficacy and safety of topical application of human fibrinogen/thrombin-coated collagen patch (TachoComb) for treatment of air leakage after standard lobectomy. AB - OBJECTIVES: Persisting air leakage after pulmonary resection remains a significant problem. The aim of the study was to evaluate the incidence of air leakage after standard lobectomy and test the efficacy and safety of TachoComb (TC). METHODS: A total of 189 patients undergoing lobectomy were enrolled in a multi-centre, open, randomised, and prospective study to test the efficacy and safety of TachoComb (TC) for air leakage treatment. Air leakage was assessed by water submersion test, and scored as grades 0 if no, 1 if countable, 2 if a stream of and 3 if coalescent bubbles have been observed. Any sites with grade 3 air leakage received further stapling or limited suturing until grade 0, 1 or 2 was obtained. Treatment of air leakage was done with TC or suturing according to randomisation. Air leakage was assessed by further submersion tests. Postoperative air leakage was assessed using the Pleur-Evac system. RESULTS: Overall incidence of air leakage 48+/-6 h after surgery was 34% for TC and 37% for standard treatment (P=0.76). The reduction of intra-operative air leak intensity in the subgroup with grades 1-2 was significantly higher for the TC group (P=0.015). Postoperative air leakage intensity in the subgroup with air leakage grades 1-2 was lower for TC than standard treatment (P=0.047). The mean duration of postoperative air leakage in the subgroup with grades 1-2 was shorter for the TC group than for standard treatment, i.e. 1.9+/-1.4 vs. 2.7+/-2.2 days (P=0.015). CONCLUSIONS: TC could be proven as well-tolerated and safe. In the subgroup of patients with established air leakage, TC showed superior potential in reduction of intra-operative air leakage as well as in reduction of intensity and duration of postoperative air leakage. PMID- 14747107 TI - Histopathological effects of ethyl 2-cyanoacrylate tissue adhesive following surgical application: an experimental study. AB - OBJECTIVES: To investigate experimentally the possible histopathological effects of ethyl 2-cyanoacrylate glue when used as a tissue adhesive in cardiovascular and thoracic surgery. METHODS: Sprague-Dawley rats were used for this study. For histopathological investigation, a study group of 144 rats in which intentionally produced lesions in myocardium (n=36), ascending aorta (n=36), lung (n=36) and abdominal aorta (n=36) were closed by using ethyl 2-cyanoacrylate was compared with the control group (n=144) in which the same lesions were closed by using sutures. On each of days 1, 7, 15, 30, 45 and 60, six rats from the study group and six rats from the control group were sacrificed and analyzed for each relevant organ in terms of bonding of ethyl 2-cyanoacrylate polymers to tissue, foreign body reaction, inflammatory reactions, and necrosis. Endothelial cell damage, intimal hyperplasia, and thrombus formation were also evaluated in arteriotomy sections. RESULTS: In histopathological analysis of vascular, myocardial and pulmonary tissue sections, there was no significant histopathological difference between conventionally sutured tissues and ethyl 2 cyanoacrylate-applied tissues. CONCLUSIONS: As no significant difference between conventional suture and ethyl 2-cyanoacrylate application was detected in terms of histopathological reactions, ethyl 2-cyanoacrylate may be considered as an alternative or adjunct to conventional techniques in controlling hemorrhage that cannot be controlled by conventional methods, in tissue repair and in the control of pulmonary air leakage, and may be used in vascular, myocardial and pulmonary surgery. PMID- 14747108 TI - Clinical predictors of surgical outcome in patients with thoracic outlet syndrome operated on via transaxillary approach. AB - OBJECTIVES: Although the transaxillary route appears to be the optimal approach in patients with thoracic outlet syndrome (TOS), the effect of clinical features on surgical outcome remains unclear. We conducted a study to determine the clinical predictors of surgical outcome in patients with TOS. METHODS: We examined the data charts of patients with TOS who underwent operation via transaxillary approach. We investigated the possible correlations between the clinical features and surgical outcomes, and analyzed the data with logistic regression model to clarify the effect of clinical features on surgical outcome. RESULTS: A total of 127 patients with a mean age of 32.1+/-10.0 years (range 14 62 years) were analyzed. The rates of favorable and poor surgical outcomes were 82.7 and 17.3%, respectively. The subgroups of symptom duration (P=0.023), the subgroups of ulnar nerve conduction velocity (P=0.033) and the presence of cervical rib (P=0.003) showed a significant correlation with surgical outcome. Multivariate analysis revealed that the shorter duration of symptoms (P=0.017) and the presence of a cervical rib (P=0.026) had a significantly unfavorable effect on surgical outcome. CONCLUSIONS: The shorter duration of symptoms and the presence of a cervical rib may imply an unfavorable surgical outcome in patients with TOS. PMID- 14747109 TI - Lobar transplantation, split lung transplantation and peripheral segmental resection--reliable procedures for downsizing donor lungs. AB - OBJECTIVES: The increasing scarcity of donor lungs, especially for small and pediatric recipients has stimulated the development of new operative techniques, which allow larger lungs to be downsized for use in smaller recipients. This approach has only recently gained widespread use-especially for highly urgent recipients-however, it is still not considered a standard procedure. METHODS: This report reviews the Vienna University experience with cadaveric split lung transplantation, lobar transplantation and by means of peripheral resection size reduced lung transplantation within the years 2001-2002. Peri-operative complications and outcome of those patients were retrospectively analysed and compared to the patients undergoing standard single or double lung transplantation during the observation period. RESULTS: During the observation period 98 primary lung transplantations were performed, of which 27 (27.6%) were size reduced transplantations. Size reduction was achieved by lobar transplantation (n=9), split lung transplantation (n=2) or peripheral segmental resection (n=16). There was no statistically significant difference between the size reduced and standard lung transplantation group with regard to the rate of bronchial healing problems (n=3/7; P=0.85) and the rate of post-operative bleeding (n=5/12; P=0.85). No other major thoracic surgical complications were observed. Three months survival rate was 85.2% in the size reduced group, compared to 92.9% in the standard group (P=0.13). CONCLUSIONS: Size reduced lung transplantation, including split lung transplantation, lobar transplantation and peripheral segmental resection, is a reliable procedure providing equal results compared to standard lung transplantation. PMID- 14747110 TI - Treatment of severe acute lung allograft rejection with OKT3 and temporary extracorporeal membrane oxygenation bridging. AB - OBJECTIVES: The use of OKT3 for treatment of advanced high-grade acute rejection episodes eventually can result in cytokine release and consecutive pulmonary edema. Temporary extracorporeal membrane oxygenation (ECMO) bridging can be used to overcome this crucial period before the beneficial effects of OKT3 can be observed. METHODS: We summarize our experience with three patients, who underwent lung transplantation and presented with severe acute rejection episodes. OKT3 had to be initiated due to insufficient response to standard rejection therapy with corticosteroids. Upon initiation of OKT3 treatment, a massive life-threatening deterioration of lung function in spite of heavily invasive respirator treatment was seen and temporary ECMO support was imperative to support graft function. Results of this treatment were retrospectively reviewed. RESULTS: In all cases femoro-femoral veno-arterial ECMO was used for support of the impaired graft and after a period of 4-5 days led to a massive improvement of graft function. In the further course two patients could be discharged from hospital and are still alive 30 and 36 months, respectively, after the described incident. One patient died 4 months later due to liver failure. CONCLUSIONS: We conclude that the use of ECMO support in patients experiencing significant side effects from OKT3 therapy is a useful and effective therapeutic tool to overcome the initial critical period until the lung has sufficiently recovered. PMID- 14747111 TI - Enhanced cytoprotection and angiogenesis by bone marrow cell transplantation may contribute to improved ischemic myocardial function. AB - OBJECTIVES: Heat shock proteins (HSPs) are cytoprotective proteins. Vascular endothelial growth factor (VEGF) is the most potent angiogenic factor. This study aimed to elucidate the possible role of cytoprotection and angiogenesis on cardiac function after bone marrow cell transplantation (BMT). METHODS: Myocardial infarction was induced in inbred Lewis rats by left anterior descending artery ligation. A total of 5 x 10(6) bone marrow-mononuclear cells were transplanted into the ischemic zone by direct injection. At 1, 3, 7, 14 and 28 days post-transplantation, cardiac function was evaluated by echocardiography. The expressions of HSP32, HSP70 and VEGF were assessed by immunofluorescence and RT-PCR. The number of vessels was examined by immunohistochemistry. The differentiation of the transplanted cells was determined by immunofluorescence. RESULTS: Echocardiography showed BMT led to sustained improvement in cardiac function, as assessed by left ventricle ejection fraction and fraction of shortening. Immunofluorescence revealed that the expressions of HSP32, HSP70 and VEGF were promoted in both transplanted bone marrow cells and recipient cardiomyocytes. RT-PCR showed that the mRNA expression levels of HSP32, HSP70 and VEGF in the BMT group were markedly higher in comparison with injection of peripheral blood cells or saline (P<0.01) by day 7. Seven days later, the vessel count showed that angiogenesis had been induced to a significantly greater degree in the BMT groups. Fourteen days later, specific markers for myocardial or vascular endothelial cells were detected in the transplanted bone marrow cells. CONCLUSIONS: BMT upregulated the expressions of HSP32, HSP70 and VEGF in both transplanted bone marrow cells and recipient endogenous cardiomyocytes in the early phase post-transplantation. This enhanced cytoprotection and angiogenesis, and contributed to the functional recovery following cardiac infarction. In the late phase, the transplanted bone marrow cells might differentiate into both myocardial and vascular endothelial cells that enhanced the ischemic cardiac function further. PMID- 14747112 TI - Cardiac complications during waiting for elective coronary artery bypass graft surgery: incidence, temporal distribution and predictive factors. AB - OBJECTIVES: Since waiting lists for coronary artery bypass graft surgery are common and carry a risk of severe events, the purposes of this study were: (1) to analyse the incidence and temporal distribution of cardiac complications during waiting for elective coronary artery bypass grafting; (2) to identify predictive factors of such complications. METHODS: Data were collected from 574 patients referred to surgery from 1 January 1998 to 12 July 2001. Two types of complications were defined: (1) a composite end-point, which included cardiac death, myocardial infarction, unstable angina or hospital admission due to cardiac cause; (2) sudden or any cardiac death. Previous cardiac events, risk factors, clinical features, laboratory exams, non-invasive tests for myocardial ischemia, left ventricular function and coronary anatomy were analysed. Kaplan Meier method, multivariate Cox regression and Student's t-test were used for statistical analyses. RESULTS: Median time to surgery was 126 days (5-1022). Among 516 patients consecutively referred to the surgery from 1 January 1998 to 31 December 2000, sudden or cardiac death occurred in 2.5% and the composite end point in 22.9%. Most complications (72.1%) were observed within 120 days. The main factors predictive of sudden or cardiac death were severe left ventricular dysfunction and heart failure (univariate analysis). Independent predictive factors of the composite end-point were angina, heart failure functional classes and high triglyceride levels. CONCLUSIONS: During long delay for coronary artery bypass surgery, cardiac events are frequent and tend to occur early. Severe left ventricular dysfunction, advanced angina, heart failure functional classes and high triglyceride level must be considered when selection is necessary, in order to diminish morbidity and mortality during the waiting period. PMID- 14747113 TI - Preoperative prediction of prolonged stay in the intensive care unit for coronary bypass surgery. AB - OBJECTIVES: To construct a predictive model for a prolonged stay in the intensive care unit (ICU) for coronary artery bypass graft surgery (CABG). METHODS: Eight hundred and eighty-eight patients undergoing CABG were studied by univariate and multivariate analysis. Prolonged stay in the ICU was defined as >/=3 days stay. Stepwise selective procedure (P/=0.40 was used as cut-off point for the prognostic test. The specificity of this test for prolonged stay in the ICU was 99%; sensitivity 9%; positive predictive value 60%; and negative predictive value 89%. CONCLUSIONS: The results show that individual patients presented for CABG, can be stratified according to their risk for prolonged stay >/=3 days in the ICU. PMID- 14747114 TI - Endothelial integrity of ultrasonically skeletonized internal thoracic artery: morphological analysis with scanning electron microscopy. AB - OBJECTIVE: The skeletonized internal thoracic artery (ITA) has several advantages over a pedicled one in coronary artery bypass grafting (CABG). An ultrasonic scalpel makes ITA skeletonization easy and speedy, however, the ultrasonic energy that is transmitted to the artery itself can occasionally injure the endothelium. Therefore, the endothelial integrity of the ultrasonically skeletonized ITA is a major concern related to this technique. The purpose of this study is to assess the endothelial integrity of the ultrasonically skeletonized ITA. METHODS: We skeletonized the left ITA with an ultrasonic scalpel in nine patients who underwent CABG, and thereafter the terminal portion of this artery was subjected to a morphological study. The endothelial integrity of this artery was morphologically assessed using scanning electron microscopy, and the results were compared to that of the left ITA skeletonized with fine scissors. RESULTS: All ITA specimens showed a completely confluent endothelium, and no endothelial injury was observed by the scanning electron microscopic study. CONCLUSIONS: The skeletonization of the ITA with an ultrasonic scalpel had no deleterious effect on the endothelium. This morphological study confirmed the safety and the reliability of this technique, and we therefore recommend its clinical use in the skeletonization of the ITA for CABG. PMID- 14747115 TI - Coronary artery bypass graft operations can be performed safely in obese patients. AB - OBJECTIVES: Obesity is a major public health problem with an increasing prevalence. Although coronary artery bypass grafting (CABG) operations are now performed with low morbidity and mortality rates, obesity is still assumed to be an important risk factor for morbidity and mortality at these operations but there is no precise approach to define it as a risk factor. The aim of this study is to evaluate the effects of obesity on the clinical outcome of the CABG operations. METHODS: A total of 1206 patients, who underwent isolated CABG operation under cardiopulmonary bypass were evaluated retrospectively. The patients were divided into three groups. Group I was normal weight, with body mass index (BMI) of 18-24.9 kg/m(2), group II was overweight, with a BMI of 25 29.9 kg/m(2), and group III was obese, with a BMI of >30 kg/m(2). The clinical data of three groups were evaluated in aspects of postoperative morbidity and mortality. RESULTS: Except for the superficial wound infections, there were no differences in postoperative mortality and morbidity rates between the three groups. Obesity was not found to be an important risk factor for postoperative morbidity and mortality. CONCLUSIONS: Despite the perception that obesity increases the risk of mortality and morbidity in CABG operations, the clinical outcome of these patients are not so different from other patients. We may say that obese patients can be safely operated. PMID- 14747116 TI - Successful treatment of deep sternal infections following open heart surgery by bilateral pectoralis major flaps. AB - OBJECTIVES: Severe sternum necrosis requiring extended resection necessitates plastic reconstruction of the resulting defect and stabilization of the chest. We analyzed the outcome of patients undergoing bilateral pectoralis major flap repair on functional and cosmetic results, chest stabilization and pulmonary function. METHODS: Twelve patients undergoing cardiac surgery between 1997 and 2001 suffered from a deep mediastinal wound infection and sternum necrosis. After a mean of two attempts of extensive wound debridement, all 12 patients underwent complete sternal resection with plastic reconstruction by bilateral pectoralis major flaps. Risk factors were obesity (n=10) and diabetes (n=11). Six months postoperatively patients underwent physical examination, pulmonary function testing and functional CT scan. RESULTS: Three patients died in hospital (two septic multiorgan failure, one heart failure) and nine were discharged with complete wound closure. One patient suffered a lethal stroke during follow-up. At 6-month follow-up no recurrent sternum infection had occurred. Chest stability was satisfactory without impairment of pulmonary function (VC 77.5+/-12.1% at follow-up vs 77.8+/-12.5% preoperatively). Mobility and force of arms and shoulder were adequate; at CT scan the maximum distance change between the former sternoclavicular joint in inspiration versus expiration was minimal. Quality of life questionnaires showed no significant limitations except a disturbed sleep and mild restriction of executing hobbies and social activities. CONCLUSIONS: Bilateral pectoralis major flap repair is a safe technique to cure severe mediastinitis necessitating complete sternal resection. Wounds close without extensive reconstructive surgery. Cosmetic results as well as stabilization of the chest were good. Patients reported an almost uncompromised quality of life without respiratory impairment. PMID- 14747117 TI - Supraventricular tachyarrythmia prophylaxis after coronary artery surgery in chronic obstructive pulmonary disease patients (early amiodarone prophylaxis trial). AB - OBJECTIVES: Supraventricular tachyarrhythmias (SVT) is common after coronary artery bypass grafting in chronic obstructive pulmonary disease (COPD). Preoperative FEV(1) is the major predetermining factor of mortality, morbidity and SVT. METHODS: Patients were divided into two groups according to their preoperative FEV(1) values. FEV(1) is <75% of predicted value in group 1 (no. 200), and >/=75% of predicted value in group 2 (no. 100). Group 1 is divided into two subgroups. SVT prophylaxis was not done in A subgroup (no. 100) whereas arrhythmia prophylaxis was done with amiodarone in all B subgroups (no. 100) in the early postoperative period. RESULTS: Atrial fibrillation developed in 28 patients in group 1A, whereas it developed in 12 in group 1B (P=0.005). Atrial flutter developed in 10 patients in group 1A but in 3 patients in group 1B (P=0.045). Multifocal atrial tachycardia developed in 13 patients in group 1A and in 4 in group 1B (P=0.022). Multivariate analysis identified ejection fraction (P<0.002, odds ratio (OR) 0.93), inotropy requirement (P<0.001, OR 3.98) amiodarone (P<0.001, OR 0.18), and FEV(1)<75% of predicted value (P<0.048, OR 1.84) as predictor of SVT. There were statistically significant differences between A and B subgroups of group 1 for hospital (P<0.001) and intensive care unit (ICU) stay (P<0.001). There was also statistically significant difference between groups 1A and 2 comparison for ICU (P<0.001; 6.4+/-3.4 versus 1.4+/-0.6 days) and hospital stay (P<0.001; 17.6+/-8.2 versus 6.9+/-0.6 days). CONCLUSIONS: Early prophylactic amiodarone not only significantly reduces SVT but also reduces SVT-related hospital and ICU stay. We strongly recommend prophylactic early use of amiodarone in COPD patients. PMID- 14747118 TI - The role of coronary angiography in acute type A aortic dissection. AB - OBJECTIVES: In recent years, non-invasive methods have replaced angiography in the diagnosis of aortic dissection. Angiography maybe used to evaluate coexisting coronary artery disease (CAD), which can delay surgery and increase the risk of rupture. We set out to examine the role of angiography in acute aortic dissection. METHODS: A retrospective analysis of patients who underwent repair of acute aortic dissection between January 1992 and June 2002 was conducted. The effect of angiography on the need for concomitant coronary artery surgery (CABG), delay to surgery and outcome were analysed. RESULTS: Seventy-four patients were identified. Initial diagnosis was established by non-invasive techniques. Twenty three patients (31%) underwent angiography (Group I) in three this was unsuccessful. Three in Group I and four in the non-angiography group (Group II, n=51) had history of angina. One patient in Group I underwent concomitant CABG compared to seven in Group II. The patient who underwent CABG in Group I; and four out of seven in Group II died (NS). Patients who underwent concomitant CABG had a significantly higher mortality rate (P=0.04). Mortality in Group I was 35% (n=8) and in Group II was 29% (n=15) (NS). Mortality rate was also significantly higher in patients who presented with syncope (P=0.01) or hypotension (P=0.04). Median transfer time from arrival at our centre to the operating room was 5 h in Group I and 1.5 h in Group II (P<0.001). Mortality rate was higher in patients who took longer to transfer to the operating room, but this did not reach statistical significance. CONCLUSIONS: We have shown that coronary angiography did not affect the occurrence of CABG and was not associated with improved hospital survival. Furthermore, there is a considerable delay to surgery caused by angiography. Therefore in this setting coronary angiography is not recommended. PMID- 14747119 TI - Can the principles of evidence-based medicine be applied to the treatment of aortic dissections? AB - Surgical treatment of patients with acute type A aortic dissections has improved early survival from 10-20 to approximately 80%. Data supporting several other treatment recommendations in patients with aortic dissection, however, are less convincing. We hypothesized that applying strict principles of evidence-based medicine would invalidate most of the recommendations in these published papers. We conducted a literature search asking three questions: (1) Is the use of routine circulatory arrest and an 'open distal' anastomosis technique better than traditional aortic cross clamping? (2) Does a persistent false lumen in the distal aorta wall have an adverse influence on long-term event-free survival? and (3) Is primary surgical or medical treatment of patients with Stanford acute type B dissections preferable in terms of long-term event-free survival? We searched Entrez Pubmed (National Library of Medicine) for all papers on these topics from 1980 to January 2003. Screening 3164 papers identified using the search terms 'aortic dissection' and 'treatment' yielded 15 papers fulfilling a set of a priori inclusion criteria. No study had a design that allowed unequivocal conclusions; moreover, the heterogeneity in study design and patient populations precluded formal meta-analysis. The difficulties inherent in conducting stringent clinical studies addressing various treatment strategies for patients with aortic dissection hamper their quality and weaken their recommendations for different treatment options. Specifically, no conclusive evidence exists favoring use of an open distal anastomosis in patients with acute type A dissections or complete elimination of flow in the distal aortic false lumen; similarly, medical therapy of patients with acute type B aortic dissections has no proven advantage over surgical treatment. PMID- 14747121 TI - Arterial switch operation with in situ coronary reallocation for transposition of great arteries with single coronary artery. AB - OBJECTIVES: Transposition of great arteries (TGA) with single coronary artery pattern is one of the high-risk groups for arterial switch operation (ASO). Any traction or kinking during coronary transfer can lead to a fatal outcome. With increase in experience, surgical results improved, but it did not completely eliminate the risks of coronary translocation. Many techniques have been described for transfer of single coronary and each one has its own merits and problems. We here in describe a new technique of in situ coronary reallocation during ASO for TGA with single coronary and also report the early and mid-term results with this new technique. METHODS: From September 1988 to June 2002, five consecutive cases of TGA with single coronary artery were operated employing this new technique. Their age ranged from 16 days to 9 months. ASO was done by transecting the great arteries just above the commissures. For coronary reallocation, hockey stick-shaped incisions were made in the facing sinuses of the proximal aorta and the pulmonary artery. These flaps were sutured in such a way that the coronary ostium was committed to the neo-aorta with the rest of surgical procedure done in the usual manner. RESULTS: All five patients had ASO. Additionally, four patients had closure of an associated ventricular septal defect and one patient had repair of the coarctation of the aorta. There was no in hospital mortality. All patients had follow-up echocardiograms at regular intervals, which showed no significant right or left ventricular outflow obstruction, no regional wall motion abnormalities and no neo-aortic or neo pulmonary regurgitation. Three of five patients had cardiac catheterization and angiocardiography, which showed normal coronary arteries with no obstructive lesions and no neo-aortic regurgitation. Their follow up ranged from 5 to 50 months and there was no late mortality. CONCLUSIONS: This new coronary reallocation technique avoids problems related to coronary translocation such as traction and kinking. It spares the need for dissection of proximal coronary artery and its branches, and thereby eliminates the risk of development of fibrosis and stenosis. The same technique can be used regardless of the sinus of origin of the coronary artery. It is a reliable and a reproducible technique. The early and mid-term results appear excellent in this series. PMID- 14747122 TI - Long-term results of correction of tetralogy of Fallot in adulthood. AB - OBJECTIVES: The natural history of tetralogy of Fallot (TOF) allows that a minority of patients reach adulthood without any treatment, representing mild forms of the disease. The aim of this study is the long-term evaluation of patients with TOF surgically treated in adulthood, in order to define its real benefit. METHODS: Between November 1982 and January 2001, 39 patients older than 18 years of age with tetralogy of Fallot underwent total correction. Mean age was 26.6 years (range 18-67) and 21 patients (53.8%) were females. A previous modified Blalock-Taussig shunt was performed in four patients (10.3%). Fifteen patients (38.5%) were in NYHA functional class III or IV. Mean hematocrit was 53.6+/-10% and the mean gradient across the right-ventricular outflow tract was 93.9+/-24.8 mmHg. The operation was performed via transatrial/transpulmonary approach in 16 patients (41%) and six patients (15.4%) required transannular patch. Pulmonary valvotomy was necessary in 13 patients (33.3%) and pulmonary valve replacement with bioprosthesis in 3 patients (7.7%). RESULTS: Hospital and late mortality were 5.1 and 7.7%, respectively. The mean follow-up was 45.1 months (range 1-194 months). Actuarial survival was 91.2+/-4.9%, 85.5+/-7.2% and 68.4+/-16.3% at 3, 7 and 15 years, respectively. In the latest follow-up, 27 (79.4%) of the survivals are presently in NYHA functional class I (P<0.001). Echocardiography has shown moderate/severe pulmonary insufficiency in 9 patients (26.5%), moderate pulmonary stenosis in 3 patients (8.8%) and residual ventricular septal defect in 4 patients (11.8%). Arrhythmias were identified in 38.9% of patients with symptoms suspicious of rhythm disturbances. There was impairment of right-ventricular function in 13 patients (38.2%). Three patients were reoperated on to close residual ventricular septal defects in two patients and for pulmonary valve replacement in one patient. CONCLUSIONS: The overall survival of surgically treated adult patients with TOF is acceptable. The great benefit of the complete repair at this age is the functional improvement. On the other hand, late complications closely related to chronic hypoxia, such as arrhythmia and ventricular dysfunction might direct for a more careful follow-up after the surgical correction. PMID- 14747123 TI - Pharmacokinetics of intravenous flucloxacillin and amoxicillin in neonatal and infant cardiopulmonary bypass surgery. AB - OBJECTIVES: To determine the blood and tissue concentrations of flucloxacillin and amoxicillin during cardiopulmonary bypass (CPB) in infants weighing less than 5 kg. METHODS: Eleven patients aged between 3 and 60 days and weighing <5 kg. Intravenous flucloxacillin 30 mg kg(-1) and amoxicillin 30 mg kg(-1) were administered at time of anaesthesia. Blood and muscle samples were collected at four stages of the operation: immediately before commencement of CPB; before cross-clamping; after weaning of CPB; and at the time of skin closure. Concentrations, half-lives (t(1/2)), clearance and volume of distribution were calculated for both antibiotics in serum and tissue. RESULTS: After connection to CPB the mean serum concentration of flucloxacillin decreased by 42.5% from 75.5 to 43.4 mg l(-1) (P=0.003) and that of amoxicillin decreased by 36.2% from 73.3 to 46.7 mg l(-1) (P=0.021). Serum concentrations of the two antibiotics decreased by a further 16.5 and 14.5% during the remainder of the surgery, but remained >15 fold above the expected minimum inhibitory concentration (MIC) for target pathogens. Muscle concentrations of both antibiotics reached MIC values by the time of the first sample and there was no decrease associated with connection to CPB. Levels remained >8-fold above MIC for target pathogens throughout the procedure. The t(1/2) for flucloxacillin was 2.64(+/-0.23)h and for amoxicillin was 3.16(+/-0.29)h, both of which are more than double the values in normal adults. There was an equivalent reduction in clearance for both antibiotics. CONCLUSIONS: Single doses of flucloxacillin and amoxicillin at 30 mg kg(-1) maintain serum and muscle concentrations well above the MIC throughout cardiac surgery. This is partly due to a prolonged t(1/2) and reduced clearance of both antibiotics in infants. PMID- 14747124 TI - Clinical evaluation of a new fat removal filter during cardiac surgery. AB - OBJECTIVES: Fat microemboli are generated during cardiac surgery that are associated with post-operative organ injury. Recently, a fat removal filter has been developed, based on a polyester leukocyte depletion filter. However, the efficacy of such a filter in a clinical setting is unknown. In this study we tested the efficacy of this filter. METHODS: Coronary artery bypass patients were randomly divided into two groups. Group I: filtration of cardiotomy suction blood during cardiopulmonary bypass with a fat removal filter (n=14). Group II: control patients without filtration (n=14). Filter efficacy was evaluated in group I using biochemical assays and thin layer chromatography of blood samples taken simultaneously before and after the filter. In addition, clinical and biochemical markers for organ injury were determined in both groups. RESULTS: The fat filter removed triglycerides (0.9+/-0.08 vs. 0.63+/-0.08 mmol l(-1), P=0.004, paired t test), leukocytes (4.3+/-0.8 x 10(9) vs. 2.3+/-0.6 x 10(9)l(-1), P=0.03), and platelets (116+/-26 x 10(9) vs. 75+/-21 x 10(9)l(-1), P=0.003) from the blood samples taken before and after the filter. Chromatography showed a significant reduction in free fatty acids, phospholipids and triglycerides. Clinically, leukocyte counts were similar, but platelet counts were higher (181+/-14 x 10(9) vs. 117+/-8.6 x 10(9)l(-1) control, P<0.001) in group I on the first postoperative day. CONCLUSIONS: The fat filter removed 40% fat, leukocytes and platelets from cardiotomy suction blood during cardiac surgery. A larger scale study is necessary to determine clinical effects on organ damage. PMID- 14747126 TI - Bacterial translocation secondary to small intestinal mucosal ischemia during cardiopulmonary bypass. Measurement by diamine oxidase and peptidoglycan. AB - OBJECTIVES: To demonstrate that small intestinal mucosal ischemia occurs during cardiopulmonary bypass by measuring serum diamine oxidase activity, an index of small intestinal mucosal ischemia, in perioerative patients undergoing cardiovascular surgery with and without cardiopulmonary bypass. METHODS: Twelve successive patients who underwent coronary artery bypass grafting with cardiopulmonary bypass (Group I) were compared to 10 patients who underwent off pump coronary artery bypass grafting (Group II). Serum diamine oxidase activity, blood lactate concentration, and serum peptidoglycan concentration were measured perioperatively. RESULTS: Serum diamine oxidase activity rose after the start of cardiopulmonary bypass and continued to rise throughout cardiopulmonary bypass in Group I, while activity was unchanged in Group II. The serum lactate concentration mirrored the change in the diamine oxidase activity in both groups. The peptidoglycan concentration in Group I rose after the start of cardiopulmonary bypass and returned to near normal concentrations after surgery. CONCLUSIONS: The parallel rise in diamine oxidase activity and the serum lactate concentration in Group I implies that ischemic injury to the mucosa of the small intestine occurs during cardiopulmonary bypass, and the rise in the serum peptidoglycan concentration indicates that bacteremia did occur. Thus, cardiopulmonary bypass causes hypoperfusion of small intestinal mucosa and consequently bacterial translocation. PMID- 14747125 TI - The effect of leucocyte-depleting arterial line filters on cerebral microemboli and neuropsychological outcome following coronary artery bypass surgery. AB - OBJECTIVES: A randomised clinical trial sought evidence as to whether leucocyte depleting (LD) arterial line filters added a further degree of neuroprotection in patients undergoing elective coronary artery bypass graft (CABG) surgery. METHODS: One hundred and ninety-two patients were randomised to the use of a Pall Leukoguard-6 LD filter or either an Avecor Affinity or Pall Autovent-6 control filter. Cerebral microemboli during surgery were recorded by transcranial Doppler (TCD) monitor over the right middle cerebral artery. Evidence of cerebral impairment was obtained by comparing patients' performance in a neuropsychological (NP) test battery (nine tests) administered 6-8 weeks post operatively with their pre-operative scores. RESULTS: The groups proved well balanced in pre-operative variables. During cardiopulmonary bypass (CPB) the median number and range of microemboli was 15 (3-180) in the LD group compared to 67 (5-846) and 55 (2-773) for the Avecor and AV6 groups, respectively (P<0.0001). One hundred and sixty-two patients completed all the NP tests. The LD group showed better post-operative performance in all but one of the nine tests although the difference in a total change score just failed to reach significance (P=0.07 one-tailed t-test). CONCLUSIONS: LD filtration during CABG reduced the number of cerebral microemboli recorded by TCD and showed a strong trend towards improving NP performance post-operatively. These findings suggest that the use of such filters in CABG surgery may offer increased neuroprotection. PMID- 14747127 TI - Mixed total anomalous pulmonary venous connection. PMID- 14747128 TI - The rare case of pulmonary artery-right inferior pulmonary vein shunt. PMID- 14747129 TI - Pseudoaneurysm of the left ventricle near the non-coronary sinus valsalvae after aortic valve replacement. PMID- 14747130 TI - Occlusion and stenosis of coronary perfusion graft after reconstruction of ascending aorta with the cabrol technique. PMID- 14747131 TI - Ascending aortic thrombus causing stroke. PMID- 14747132 TI - Tracheal release and thymus wrapping of the tracheoplasty anastomosis through mini-sternotomy. AB - Despite breakthroughs in general thoracic surgery, tracheoplasty remains a technically difficult procedure. The authors performed tracheoplasty on a 75-year old woman diagnosed with recurrent thyroid cancer and tracheal invasion. Through an L-shaped unilateral mini-sternotomy added to a collar incision, we performed tracheal mobilization and release followed by resection of six tracheal rings. The tracheoplasty anastomosis was then wrapped with the right lobe of the thymus. Using this procedure, tracheoplasty was successfully performed with no complications related to the tracheal anastomosis. Tracheoplasty performed through mini-sternotomy, with a combination of full tracheal release and wrapping with the right lobe of the thymus, may represent a novel, effective, and less invasive surgical procedure. PMID- 14747133 TI - Right parasternal approach for aortic valve replacement after retrosternal gastropexy. AB - Aortic valve replacement had to be performed in a 77-year-old man with a history of esophageal carcinoma, which had been treated with two-staged esophageal resection and retrosternal gastropexy. Barium swallowing confirmed the retrosternal course of the stomach, which crossed the midline from the right upper abdomen to the left-sided neck anastomosis. Aortic valve replacement with a bioprosthesis was performed through a small right parasternal thoracotomy. The postoperative course was uneventful. We found that the right parasternal incision is an excellent surgical approach for aortic valve replacement in patients after retrosternal gastropexy. PMID- 14747134 TI - Surgical one-stage approach for coronary artery disease and occlusive disease of all aortic arch branches. AB - A 52-year-old man, who presented with two-vessel coronary artery disease and severe arterial occlusive disease with occlusions and/or stenoses of all aortic arch branches, underwent simultaneously coronary artery bypass grafting and bilateral aortic-subclavian as well as left-sided aortic-carotidal bypass grafting. PMID- 14747135 TI - Can we avert the need for pneumonectomy by screening for lung cancer? PMID- 14747136 TI - Radiation therapy alone as an alternative to surgery in patients with early stage non-small-cell lung cancer having cardiovascular (and other) comorbidity. PMID- 14747137 TI - var genes, PfEMP1 and the human host. AB - Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1) is an important virulence factor encoded by a family of roughly 60 var genes and is used by the parasite to interact with the human host. The parasite regularly exchanges the expressed var gene generating antigenic variation of the infected RBCs (pRBC) surface which is crucial for successful proliferation and transmission. PfEMP1 is also an adhesive molecule that binds to an array of human receptors. By sequestration in the post-capillary venules, pRBCs are able to escape the spleen mediated clearance but severe malaria may develop if the local binding is extensive. Anti-PfEMP1 immunity is important for preventing the development of both cerebral malaria and placental malaria, but more immunological studies on PfEMP1 antigens and their interaction with the human host are needed. Over the last few years our knowledge about var genes and PfEMP1s has increased dramatically through genetic, biochemical, immunological and epidemiological studies. In addition, the genome sequence has also provided us with a new platform for further dissecting its biological functions. This review highlights the recent analyses of var genes in the P. falciparum genome and postulates significance of genome recombination to the diversity of parasite virulence. PMID- 14747138 TI - STEVOR--a multifunctional protein? AB - The stevor multigene family is the third largest identified in Plasmodium falciparum. Its members have the potential to be involved in antigenic variation and virulence by analogy with the var and rif multigene families. This review highlights recent studies of stevor transcription and expression which show that stevor is distinct from both the var and rif multigene families. STEVOR is expressed during several stages of the lifecycle, and thus may contribute significantly to the long term survival of the parasite. PMID- 14747139 TI - Variant antigen expression in malaria infections: posttranscriptional gene silencing, virulence and severe pathology. PMID- 14747140 TI - Antigenic variation and cytoadhesion in Babesia bovis and Plasmodium falciparum: different logics achieve the same goal. AB - Babesia bovis is a protozoal hemoparasite of cattle which behaves in certain crucial respects like Plasmodium falciparum, despite being phylogenetically distant and having many differences in its life cycle. The shared behavioral attributes of rapid antigenic variation and cytoadhesion/sequestration are thought to contribute significantly to immune evasion, establishment of persistent infections, and disease pathology. Although differing in their genetic and biochemical strategies for achieving these behaviors, information from studies of each parasite may further our understanding of the overall host parasite interaction. In this review we contrast the molecular basis and 'genetic logic' for these critical behaviors in the two parasites, with emphasis on the biology of B. bovis. PMID- 14747141 TI - Meeting report: the molecular background of severe and complicated malaria. PMID- 14747142 TI - Subcellular localization of adenine and xanthine phosphoribosyltransferases in Leishmania donovani. AB - The subcellular location of a protein is a critical factor in its physiological function and an important consideration in therapeutic paradigms that target the protein. Because Leishmania donovani cannot synthesize purine nucleotides de novo, they rely predominantly upon therapeutically germane phosphoribosyltransferase (PRT) enzymes, hypoxanthine-guanine PRT (HGPRT), adenine PRT (APRT), and xanthine PRT (XPRT), for purine acquisition from the host. Previous studies have shown that the L. donovani HGPRT is localized to the glycosome, a fuel-metabolizing microbody that is unique to kinetoplastid parasites [J. Biol. Chem. 273 (1998) 1534]. The sequences of the other two PRTs indicate that XPRT, but not APRT, possesses a COOH-terminal tripeptide that mediates protein targeting to the glycosome. To determine definitively the intracellular milieu of APRT and XPRT, polyclonal antibodies were raised to each recombinant protein. APRT and XPRT were then shown by immunofluorescence to be localized to the cytosol and glycosome, respectively. The glycosomal milieu for XPRT was also verified by immunoelectron microscopy. Amputation of the glycosomal targeting signal from XPRT resulted in protein mislocalization to the cytosol, but the cytosolic xprt was still functional with respect to purine salvage. These studies establish that APRT is cytosolic and XPRT, like the homologous HGPRT, is glycosomal and demonstrate that a mutant xprt protein that mislocalizes to the cytosol is still functional and supports parasite viability. PMID- 14747143 TI - Chromatin diminution leads to rapid evolutionary changes in the organization of the germ line genomes of the parasitic nematodes A. suum and P. univalens. AB - Chromatin diminution in the parasitic nematodes Ascaris suum and Parascaris univalens represents a rather complex molecular phenomenon that includes chromosomal breakage, DNA degradation and new telomere formation. At a given elimination site, DNA breakage and new telomere addition does not take place at a single chromosomal locus but at many different places within a several kilobase long chromosomal region, referred to as chromosomal breakage region (CBR). Here we describe the cloning and the characterisation of seven CBRs from A. suum and P. univalens and we show that the process has been conserved between the two species. A detailed sequence comparison provides evidence that the sequences of the CBRs and their flanking regions are not directly important for the specification of the elimination sites. Six out of the seven CBRs are conserved between the two nematode species, suggesting that they have already existed in a common ancestor. We present a hypothesis stating that the elimination process ensures the maintenance of a functional somatic genome and concomitantly allows extremely rapid and profound changes in the germ line genome, thereby allowing the development of new germ line specific functions and thus providing a selective advantage for the chromatin eliminating nematodes during further evolution. PMID- 14747144 TI - The N-terminus moiety of the cystatin SmCys from Schistosoma mansoni regulates its inhibitory activity in vitro and in vivo. AB - The complete sequence of SmCys, a cystatin expressed by Schistosoma mansoni, was obtained. Constructs of SmCys consisting of deletions of 10 and 20 amino acid residues from the N-terminal of the full length recombinant protein, were cloned in the pQE-30 vector, expressed in Escherichia coli and assayed for inhibitory activity against papain. Kinetic analysis showed that SmCys -10 and SmCys -20 had K(i) values of 0.7391 and 4.9154, respectively, as compared to 0.0647, displayed by the full length recombinant. Protease inhibition by SmCys was also observed in vivo. When the recombinant products were incubated during 7 days with live schistosomula in the presence of red blood cells, only the full length product could completely inhibit the formation of haemozoin, a dark pigment formed as a by-product of haemoglobin digestion. The sequence data of the recombinant SmCys proteins were used for the construction of molecular models, which were then subjected to molecular dynamics for 2ns. In comparison to the full length, the models corresponding to the truncated constructs, showed a distinctive change on the surface charge distribution. This parameter was more pronounced in SmCys -20, which also displayed a significant displacement of the inhibitory domain, a result which could explain the kinetic data in terms of the loss of attachment sites. These changes correlated well with the progressive lack of inhibition observed for the recombinant deletion constructs, in vitro and in vivo. PMID- 14747145 TI - Identification of regulatory elements in the Plasmodium falciparum genome. AB - There is little information regarding regulatory sequences in the newly sequenced genome of the malaria parasite, Plasmodium falciparum. Thus, for the first time, a bioinformatic strategy was utilized to identify regulatory elements in this genome using the P. falciparum heat shock protein (hsp) gene family as a model system. Our analysis indicates that the P. falciparum hsp genes do not contain standard eukaryotic regulatory elements. However, a novel G-rich regulatory element named the G-box was identified upstream of several P. falciparum hsp genes and the P. yoelii yoelii, P. berghei, and P. vivax hsp86 genes. Remarkably, the Plasmodium sp. G-boxes were required for maximal reporter gene expression in transient transfection assays. The G-box is not homologous to known eukaryotic elements, and is the best-defined functional element elucidated from Plasmodium sp. Our analysis also revealed several other elements necessary for reporter gene expression including an upstream sequence element, the region surrounding the transcription start site, and the 5' and 3' untranslated regions. These data demonstrate that unique regulatory elements are conserved in the genomes of Plasmodium sp., and demonstrate the feasibility of bioinformatic approaches for their identification. PMID- 14747146 TI - P-glycoprotein inhibitors modulate accumulation and efflux of xenobiotics in extra and intracellular Toxoplasma gondii. AB - We examined xenobiotic transport and the effects of P-glycoprotein (Pgp) inhibitors on efflux function in Toxoplasma gondii tachyzoites. The fluorescence emission of JC-1 and daunorubicin (Pgp substrates) was determined in both extracellular tachyzoites and T. gondii-infected human KB cells. Dye accumulation and efflux were modulated by verapamil (Vp) and cyclosporin A (CsA), both of which are Pgp inhibitors. Red JC-1 emission was measured from 10(6) extracellular tachyzoites, using spectrofluorometry. The increase in red emission was significant from 1 microM concentration of both drugs and was higher with CsA than with Vp. Compared with untreated tachyzoites, JC-1 efflux was inhibited by 3 microM CsA and 3 microM Vp. With intracellular tachyzoites, the fluorescence distribution of daunorubicin (DNR) between the parasitophorous vacuole and the host cell was modulated by Vp and CsA. In media free of CsA and Vp, DNR emission inside intracellular tachyzoites was very weak, as observed by confocal microscopy. In the presence of CsA or Vp, DNR emission was markedly enhanced in tachyzoites but not in the whole vacuole. The modulation of DNR uptake seems to involve the tachyzoite membrane rather than the parasitophorous vacuole or host cell membranes. It suggests that Vp would inhibit the DNR efflux from intracellular tachyzoites through a transitory effect. In conclusion, these two Pgp inhibitors increase both extracellular and intracellular dye accumulation in living T. gondii, pointing to the existence of a transmembrane transport mediated by a Pgp homologue located on the parasite membrane complex. PMID- 14747147 TI - New selectable markers and single crossover integration for the highly versatile Plasmodium knowlesi transfection system. AB - Plasmodium knowlesi provides a highly versatile transfection system for malaria, since it enables rapid genetic modification of the parasite both in vivo as well as in vitro. However, it is not possible to perform multiple genetic manipulations within one parasite line because of a lack of selectable markers. In an effort to develop additional selectable markers for this parasite, positive and negative selectable markers that have recently been successfully used in Plasmodium falciparum were tested. It was shown that the positive selectable markers human dihydrofolate reductase (hdhfr), blasticidin S deaminase (bsd) and neomycin phosphotransferase II (neo) all conferred drug resistance to P. knowlesi when introduced as episomes. The plasmid containing the hdhfr selectable marker was not only successfully introduced as circular form, but also as linear fragment, demonstrating for the first time single crossover integration in P. knowlesi. Thymidine kinase was tested for its potential as negative selectable marker and it was shown that recombinant P. knowlesi parasites expressing thymidine kinase from episomes were highly sensitive to ganciclovir compared to wild-type P. knowlesi. The availability of new positive selectable markers and a strong candidate for a negative selectable marker for P. knowlesi, in combination with the opportunity to perform targeted single crossover integration in P. knowlesi, significantly increases the flexibility of this transfection system, making it one of the most versatile systems available for Plasmodium. PMID- 14747148 TI - A novel form of actin in Leishmania: molecular characterisation, subcellular localisation and association with subpellicular microtubules. AB - To study the occurrence and subcellular distribution of actin in trypanosomatid parasites, we have cloned and overexpressed Leishmania donovani actin gene in bacteria, purified the protein, and employed the affinity purified rabbit polyclonal anti-recombinant actin antibodies as a probe to study the organisation and subcellular distribution of actin in Leishmania cells. The Leishmania actin did not cross react with antimammalian actin antibodies but was readily recognized by the anti-Leishmania actin antibodies in both the promastigote and amastigote forms of the parasite. About 10(6) copies per cell of this protein (M(r) 42.05 kDa) were present in the Leishmania promastigote. Unlike other eukaryotic actins, the oligomeric forms of Leishmania actin were not stained by phalloidin nor were dissociated by actin filament-disrupting agents, like Latrunculin B and Cytochalasin D. Analysis of the primary structure of this protein revealed that these unusual characteristics may be related to the presence of highly diverged amino acids in the DNase I-binding loop (amino acids 40-50) and the hydrophobic plug (amino acids 262-272) regions of Leishmania actin. The subcellular distribution of actin was studied in the Leishmania promastigotes by employing immunoelectron and immunofluorescence microscopies. This protein was present not only in the flagella, flagellar pocket, nucleus and the kinetoplast but it was also localized on the nuclear, vacuolar and cytoplasmic face of the plasma membranes. Further, the plasma membrane-associated actin was colocalised with subpellicular microtubules, while most of the actin present in the kinetoplast colocalised with the k-DNA network. These results clearly indicate that Leishmania contains a novel form of actin which may structurally and functionally differ from other eukaryotic actins. The functional significance of these observations is discussed. PMID- 14747149 TI - Chondroitin sulfate proteoglycans of the endothelia of human umbilical vein and arteries and assessment for the adherence of Plasmodium falciparum-infected erythrocytes. AB - Infection with Plasmodium falciparum during pregnancy leads to chondroitin 4 sulfate-mediated adhesion of the infected red blood cells (IRBCs) in the placenta, causing severe health complications to fetus and the mother. The IRBCs are also frequently found in low density in the umbilical cord of infected placentas. In this study, the CSPGs of umbilical vein and arteries were purified, characterized, and their localization and IRBC-binding abilities were studied. While a versican type CSPG was found both in the vein and arteries, a serglycin type CSPG was present exclusively in the vein. The CSPGs were present at significant level on the endothelial surface of the umbilical vein but not on that of arteries. Although the purified versican and serglycin type CSPGs could bind IRBCs, their binding abilities were significantly less compared to the low sulfated CSPGs of the placenta because of the predominance of 6-sulfated disaccharide moieties in the CS chains. Therefore, IRBCs were unable to bind efficiently onto the umbilical cord endothelial surface. Unexpectedly, however, the IRBCs adhered densely in the blood vessels of fetal villi in the placental tissue sections and sparingly in the blood spaces of the umbilical cord vein, presumably because the CSPG that can efficiently bind IRBCs is present at high levels in the fetal blood vessels and at very low levels in the umbilical cord blood vessels. Since the C4S-adherent IRBCs that enter the fetal blood vessels cannot adhere to the cord endothelial surface and parasites cannot efficiently grow due to fetal hemoglobin toxicity and protection by maternal antibodies, transplacental infection may be quickly cleared without clinical episodes. PMID- 14747150 TI - Expression and functional characterization of a giant Type I fatty acid synthase (CpFAS1) gene from Cryptosporidium parvum. AB - A 25-kb CpFAS1 gene from Cryptosporidium parvum has been engineered and expressed as five individual maltose-binding protein (MBP)-fusion proteins: an N-terminal loading unit, three fatty acyl elongation modules, and a C-terminal reductase. Enzymatic activities of all domains (except the reductase) were individually assayed as recombinant proteins. The preferred substrate for the fatty acyl ligase (AL) domain in the loading unit was palmitic acid (C16:0). However, a competition assay suggests that the AL domain could also utilize other fatty acids as substrates (i.e., C12:0-C24:0), albeit with reduced activity. Among the three elongation modules, enzymatic activities were detected for ketoacyl synthase (KS), acyl transferase (AT), dehydrase (DH), enoyl reductase (ER), and ketoacyl reductase (KR) domains, which suggests that these modules were involved in the elongation of a saturated fatty acyl chain that would be C6 longer (e.g., C22:0) than the precursor (e.g., C16:0). In addition, the KS activity could be specifically inhibited by cerulenin (IC(50) approximately 1.5 microM), reinforcing the notion that CpFAS1 could be exploited as potential drug target. Since C. parvum lacks other fatty acid synthases, these observations imply that this parasite may not be capable of synthesizing fatty acids de novo. PMID- 14747151 TI - A new modular protein of Cryptosporidium parvum, with ricin B and LCCL domains, expressed in the sporozoite invasive stage. AB - The recombinant SA35 peptide has been described as an antigenic portion of a larger Cryptosporidium parvum protein. We identified and characterized the encoding Cpa135 gene and the entire protein, Cpa135. The Cpa135 gene was found to consist of a single exon of 4671 bp, and the mRNA transcribed in the sporozoites was identified. The predicted 1556 amino-acid protein showed the presence of domains which are widely conserved also in other unrelated phylogenetic groups (i.e. a ricin B and a LCCL motif). Comparison of Cpa135 sequence with genomic and protein databases revealed many related genes in other apicomplexan species and high homology with CCP2 protein from Plasmodium yoelii and Plasmodium berghei. The Cpa135 protein was identified and localized by using a monoclonal antibody (Mab) directed against the SA35 antigen (anti-SA35). In oocyst-sporozoite lysate, the anti-SA35 MAb recognized a 135 kDa protein that forms a protein complex larger than 200 kDa, which is mediated by disulfide bridges. Cpa135 synthesis was up-regulated during the excystation process. After host-cell invasion, Cpa135 gene expression was undetectable up to 48 h, whereas mRNA synthesis was newly observed at 72 h post-infection. The Cpa135 protein was localized in the apical complex, and it was found to be secreted by sporozoites during their gliding. Cpa135 persisted during the intracellular stages of the parasite, and it defined the boundaries of the parasitophorous vacuole in the infected cells. The unique array of domains and the homology with other apicomplexan proteins indicate that the Cpa135 protein is representative of a new family of proteins. PMID- 14747152 TI - Identification and characterization of PDZ-1, a N-ERMAD specific interaction partner of the Echinococcus multilocularis ERM protein Elp. PMID- 14747153 TI - Distribution of apolipoprotein L-I and trypanosome lytic activity among primate sera. PMID- 14747154 TI - Trypanosoma (Nannomonas) simiae and T. (N.) godfreyi have genes encoding glutamic acid and alanine-rich proteins. PMID- 14747155 TI - Integration of pZJM library plasmids into unexpected locations in the Trypanosoma brucei genome. PMID- 14747156 TI - In vivo transcription analysis utilizing chromatin immunoprecipation reveals a role for trypanosome transcription factor PBP-1 in RNA polymerase III-dependent transcription. PMID- 14747157 TI - The use of DsRED in single- and dual-color fluorescence labeling of mitochondrial and plastid organelles in Plasmodium falciparum. PMID- 14747158 TI - ICSI versus conventional techniques for oocyte insemination during IVF in patients with non-male factor subfertility: a Cochrane review. AB - This paper is based on a Cochrane review of the same title by the same authors published in The Cochrane Library, issue 3, 2003 (see www.CochraneLibrary.net for information) with permission from the Cochrane Collaboration-John Wiley and Sons. Cochrane reviews are regularly updated as new evidence emerges and in response to comments and criticisms, and The Cochrane Library should be consulted for the most recent version of the review. BACKGROUND: The objective of this review was to investigate whether ICSI improves live-birth rate in comparison with IVF in couples with non-male factor subfertility. METHODS: We searched the Cochrane Menstrual Disorders and Subfertility Group trials register (searched 30 May 2002), the Cochrane Controlled Trials Register (Cochrane Library Issue 2, 2002), PubMed (January 1992 to September 2003) and reference lists of articles. Trials were included if they compared the effects of ICSI and IVF on live births, pregnancy and fertilization outcomes. Only randomized studies were included in this review. Two reviewers extracted data independently. RESULTS: There were no randomized data comparing live-birth rates. The single identified study did not find a difference in pregnancy rates (OR 1.4, 95% CI 0.95-2.2). There were no randomized data on miscarriage rates, or on other adverse events such as congenital malformations that may be of concern (415 couples randomized). Two studies used alternation to assign their couples and did have live birth as an outcome. These studies showed a significantly higher fertilization rate in the IVF group, but no difference in pregnancy, miscarriage or live-birth rate. CONCLUSIONS: Whether ICSI should be preferred to IVF for cases of non-male factor subfertility remains an open question. Further research should focus on live birth rates and adverse events. PMID- 14747159 TI - High frequency of defective sperm-zona pellucida interaction in oligozoospermic infertile men. AB - BACKGROUND: The ability of sperm to interact with the zona pellucida (ZP) plays a critical role during the process of human fertilization. The aim of this study is to determine frequency of defective sperm-ZP interaction in oligozoospermic infertile men. METHODS: Sperm-ZP binding assays and the ZP-induced acrosome reaction (AR) were performed in 72 infertile men with a sperm concentration <20 x 10(6)/ml. Oocytes that had previously failed to fertilize in a clinical IVF programme were used for the tests. Motile sperm (2 x 10(6)/ml) selected by swim up from each semen sample were incubated with four oocytes for 2 h. The number of sperm bound per ZP and the ZP-induced AR were assessed. Under these conditions, an average of < or =40 sperm bound/ZP was defined as low sperm-ZP binding and a ZP-induced AR < or =16% was defined as low ZP-induced AR. RESULTS: In the 72 oligozoospermic men, 28% (20/72) had low sperm-ZP binding. Of those with normal sperm-ZP binding, 69% (36/52) had low ZP-induced AR. Overall, 78% (56/72) had either low ZP-binding or normal ZP binding but low ZP-induced AR. This means that only 22% (16/72) had both normal sperm-ZP binding and normal ZP-induced AR. CONCLUSION: Oligozoospermic men have a very high frequency of defective sperm-ZP interaction, consistent with their low natural fertility or low fertilization rate in conventional IVF. Infertile couples with oligozoospermic semen should be treated by ICSI rather than by conventional IVF. PMID- 14747160 TI - Sperm membrane protein profiles of fertile and infertile men: identification and characterization of fertility-associated sperm antigen. AB - BACKGROUND: Male infertility is the major cause of conception failure in about 25% of all infertile couples. Understanding the causes of male infertility depends to a certain extent on the proteins present on the spermatozoa. The study aim was to investigate first, whether there is any difference in the expression of sperm membrane proteins between fertile and infertile males; and second, whether there is any functional significance of these proteins in the spermatozoa. METHODS: Six different protocols were employed to extract sperm membrane proteins. A 57 kDa protein was identified and purified using different chromatographic techniques. The homogeneity and isoelectric point of the protein was confirmed by 2D-electrophoresis. The protein was characterized by immunofluorescence, ELISA, flow cytometry, SDS-PAGE and Western blot analysis. The role of 57 kDa protein in sperm-oocyte binding was studied in vitro. RESULTS: All six sperm extracts of normozoospermic and infertile subjects showed 16-18 major and 12-15 minor protein bands. However, in one of the methods, the lysis buffer containing N-octyl-beta-D-glycopyranoside (NOG) resulted in an additional protein band at the 57 kDa region in 95% of normozoospermic samples. The protein was either absent (approximately 80%) or negligible (approximately 20%) in infertile subjects. The protein was localized to the head of non-acrosome-reacted spermatozoa (NAR), and shifted to the equatorial segment in acrosome-reacted (AR) spermatozoa. The antibody directed against purified 57 kDa protein inhibited binding of human sperm to zona-free hamster oocytes in a dose-dependent manner. CONCLUSIONS: The results suggest that lack and/or low expression of 57 kDa protein may be one of the reasons for infertility in men. Therefore, the protein could be used as a marker for sperm quality in men. PMID- 14747161 TI - SPRASA, a novel sperm protein involved in immune-mediated infertility. AB - BACKGROUND: Antisperm antibodies (ASA) may be an important cause of infertility, but current tests for the detection of ASA have poor prognostic value. Identification of the sperm proteins that ASA bind to may aid the development of more useful diagnostic tests. METHODS: One- and two-dimensional PAGE and western blotting analyses, as well as amino acid sequencing, were used to identify a novel sperm protein reactive with ASA (SPRASA) from infertile men. An antiserum reactive with SPRASA was produced by immunizing a rabbit with SPRASA excised from two-dimensional gels. This antiserum was used to demonstrate the localization of SPRASA on the sperm. RESULTS: Amino acid sequences derived from SPRASA matched those of a theoretical protein, XP-085564. This protein is derived from the C type lysozyme/alpha-lactalbumin gene family. Immunohisto chemistry indicates that SPRASA is localized to the acrosome. Western blot analysis revealed that 50 unselected individuals did not have antibodies that reacted with SPRASA. CONCLUSION: Only ASA from infertile men react with SPRASA, suggesting that this novel protein may be important in the processes of fertility. The identification of SPRASA as the antigen for infertility-associated ASA raises the possibility of developing first, antigen-specific tests for ASA, and secondly, more targeted treatment for immune-mediated infertility. PMID- 14747162 TI - Cystic fibrosis transmembrane conductance regulator gene screening and clinical correlation in Taiwanese males with congenital bilateral absence of the vas deferens. AB - BACKGROUND: In Taiwan, an area with a very low incidence of cystic fibrosis (CF), we first screened for the most common mutations of the cystic fibrosis transmembrane conductance regulator (CFTR) gene and looked for clinical correlations in 27 patients with clinically diagnosed congenital bilateral absence of the vas deferens (CBAVD). METHODS AND RESULTS: The clinical results showed that none of the 27 patients had CF symptoms. We did not detect any definite renal anomaly ultrasonographically. Mutation analysis was carried out on these 27 cases and 46 normal fertile males as controls. No mutations of Delta F508 or R117H were identified in any of the samples analysed. In the screening of IVS8-poly T, five of the 27 CBAVD patients showed the homozygous genotype for 5T/5T, 14 showed the heterozygous genotype for 5T/7T and eight showed the homozygous genotype for 7T/7T. The frequency of 5T alleles was 44.4%, which was significantly higher than in the 46 normal fertile males, for which there was a 5T frequency of 5.4%. CONCLUSIONS: The absence of major mutations of CFTR genes could be related to the much lower CF incidence in Taiwan. Further investigations into differences in the mutation spectrum of other CFTR genes are needed for a better understanding of the development of Taiwanese-Oriental CBAVD. PMID- 14747163 TI - Caspase-dependent and -independent DNA fragmentation in Sertoli and germ cells from men with primary testicular failure: relationship with histological diagnosis. AB - BACKGROUND: Germ cell elimination and sperm DNA fragmentation in men with primary testiculopathies involve apoptosis-related processes whose mechanisms are poorly understood. This study examines the participation of typical (caspase-dependent) and atypical (caspase-independent) pathways in these processes. METHODS: Caspase activity and DNA fragmentation were evaluated in Sertoli and germ cells from 63 men with non-obstructive azoospermia and with different histological diagnoses who were undergoing testicular biopsy for an assisted reproduction attempt. In eight of these men, phosphatidylserine externalization was also examined. RESULTS: The percentage of Sertoli cells showing caspase activity and DNA fragmentation was low and uniform in all diagnoses. In germ cells that remained tightly associated with Sertoli cells despite vigorous mechanical treatment, the incidence of both caspase activity and DNA fragmentation was high, particularly in men with maturation arrest. In Sertoli cell-free germ cells, high incidence of DNA fragmentation contrasted with low incidence of caspase activity and phosphatidylserine externalization. CONCLUSIONS: In men with primary testicular failure, apoptosis of Sertoli cells is insignificant. Some germ cells undergo caspase-dependent apoptosis, show phosphatidylserine externalization and are tightly associated with Sertoli cells. Other germ cells show caspase-independent DNA fragmentation, do not externalize phosphatidylserine and lack a tight association with Sertoli cells. PMID- 14747164 TI - Pregnancy with frozen-thawed and fresh testicular biopsy after motile and immotile sperm microinjection, using the mechanical touch technique to assess viability. AB - BACKGROUND: There are few reports of pregnancy using immotile sperm, and none using a purely mechanical assessment of viability. METHODS: In this pilot study, we retrospectively analysed 66 cycles in 61 patients with determinant male factor, recording rates of fertilization, implantation, normal pregnancy and take home babies achieved with ICSI. Sperm selection was based on morphologically normal appearance under the inverted microscope. Viability of immotile spermatozoa was assessed by the mechanical touch technique to observe tail flexibility and tail shape recovery. RESULTS: Of 17 ICSI cycles using frozen thawed testicular sperm, six microinjected with immotile and 11 with motile sperm, we achieved fertilization rates of 65.7 and 74.3%, respectively, and five pregnancies (two and three, respectively). Of 49 ICSI cycles using fresh testicular sperm, 10 microinjected with immotile and 39 with motile sperm, we achieved fertilization rates of 73.4 and 64.4%, respectively, and 12 pregnancies (three and nine, respectively). CONCLUSIONS: Immotile (fresh and frozen-thawed) testicular sperm of normal morphological appearance can be used to achieve clinical pregnancy with ICSI. Our results strongly suggest that immotile sperm viability can be assessed by the mechanical touch technique. PMID- 14747165 TI - A randomized controlled trial comparing medical and expectant management of first trimester miscarriage. AB - BACKGROUND: We aimed to determine whether outpatient treatment of miscarriage with vaginal misoprostol is more effective than expectant management in reducing the need for surgical evacuation of retained products of conception (ERPC). METHODS: Of 131 eligible women with first trimester miscarriage, 104 agreed to randomization to either 600 microg misoprostol or placebo intravaginally. They were assessed the following day and administered a second dose of their allocated treatment if miscarriage was not complete. Those not successful after two doses were seen on day 7, and, if miscarriage was not complete, an ERPC was performed. RESULTS: The success rate of medical management was 88.5% (46/52) compared with 44.2% (23/52) for expectant management. There was no significant difference in success rate (100 versus 85.7%) in women treated with an incomplete miscarriage. Women with early pregnancy failure had a success rate of 87% with misoprostol compared with 29% with expectant management [odds ratio (OR) 15.96; 95% confidence interval (CI) 5.26, 48.37]. The complete miscarriage rate was achieved quicker in the medical group than the expectant group by day 1 (32.7 versus 5.8%) and by day 2 (73.1 versus 13.5%) of treatment. There were no differences in side effects, bleeding duration, analgesia use, pain score and satisfaction with treatment. Women in the expectant group made more outpatient visits (5.06 versus 4.44%; OR = -0.62, 95% CI -1.04, -0.19). More women in the medical group (90.4 versus 73.1%; OR 1.26, 95% CI 1.05, 1.50) would elect the same treatment in the future. CONCLUSIONS: Medical management using 600 microg misoprostol vaginally is more effective than expectant management of early pregnancy failure. Misoprostol did not increase the side-effect profile and patient acceptability was superior to expectant management. PMID- 14747166 TI - Incidence of spontaneous abortion among pregnancies produced by assisted reproductive technology. AB - BACKGROUND: There has been increasing number of pregnancies following assisted reproductive technology treatment and their survival is understandably a matter of interest. The relative risk of spontaneous abortion in these pregnancies remains unclear. The objectives of this study were to quantify the relative risk in assisted reproductive technology pregnancies in relation to two cohorts of naturally conceived pregnancies and to assess the possible risk factors for spontaneous abortion among assisted reproductive technology pregnancies. METHODS: Three cohorts of pregnancies, 1945 pregnancies conceived following assisted reproductive technology treatment in a tertiary infertility clinic, 549 natural pregnancies in a prospective study of lifestyle and pregnancy (the Ford cohort), and 4265 pregnancies from another cohort (the Treloar cohort), were used in the study. RESULTS: After adjusting for age, the relative risk of spontaneous abortion was 1.20 (95% CI 1.03-1.46) in the assisted reproductive technology cohort in comparison with the Ford cohort. Within the assisted reproductive technology cohort, a history of spontaneous abortion predicted increased risk, while a low level of ovarian stimulation seemed to be related to a reduced risk. CONCLUSIONS: The study showed that the risk of spontaneous abortion was slightly increased in the assisted reproductive technology pregnancies after adjusting for maternal age and previous spontaneous abortion. Within the assisted reproductive technology cohort, several variables, including the level of stimulation, appeared to be linked with the risk of spontaneous abortion. PMID- 14747167 TI - Ectopic pregnancies in Caesarean section scars: the 8 year experience of one medical centre. AB - BACKGROUND: Our aim was to supplement the mostly individual case reports on the rarely occurring and life-threatening condition of ectopic pregnancy developing in a Caesarean section scar. METHODS AND RESULTS: Eight of all the patients treated in our department between 1995 and 2002 had been diagnosed for ectopic pregnancy that developed in a Caesarean section scar. They comprised this case series group. Four of them underwent methotrexate treatment; one had expectant management, one transcervical aspiration of the gestational sac and two by open surgery. All the non-surgically treated women had an uneventful outcome. One underwent a term Caesarean hysterectomy and the other first trimester hysterotomy and excision of the pregnancy located in the scarred uterus. Analysis of all these women's obstetric history revealed that five of them (63%) had been previously operated because of breech presentation, one had a cervical pregnancy and one had placenta previa. Four of them (50%) had multiple (> or = 2) Caesarean sections. CONCLUSIONS: The women at risk for pregnancy in a Caesarean section scar appear to be those with a history of placental pathology, ectopic pregnancy, multiple Caesarean sections and Caesarean breech delivery. Heightened awareness of this possibility and early diagnosis by means of transvaginal sonography can improve outcome and minimize the need for emergency extended surgery. PMID- 14747168 TI - Heterotopic Caesarean scar pregnancy combined with intrauterine pregnancy successfully treated with embryo aspiration for selective embryo reduction: case report. AB - Ectopic pregnancy situated in a Caesarean section scar is a rare but potentially life-threatening event. Because of its rarity, there are no universal treatment guidelines to manage this condition. We report a case of IVF-induced triplet heterotopic pregnancy of early gestational age that included one Caesarean scar pregnancy diagnosed as early as 6 weeks gestation. Treatment with embryo aspiration under vaginal ultrasonography for selective embryo reduction was given and the concurrent intrauterine twin pregnancy was preserved successfully. PMID- 14747169 TI - Computer-controlled, multilevel, morphometric analysis of blastomere size as biomarker of fragmentation and multinuclearity in human embryos. AB - BACKGROUND: Little is known about blastomere size at different cleavage stages and its correlation with embryo quality in human embryos. Using a computer system for multilevel embryo morphology analysis we have analysed blastomeres of human embryos and correlated mean blastomere size with embryonic fragmentation and multinuclearity. METHODS: A consecutive cohort of 232 human 2-, 3- and 4-cell embryos from patients referred for ICSI treatment were included. Sequences of digital images were taken by focusing at 5- micro m intervals through the embryo. Blastomere sizes and number of nuclear structures were evaluated based on these sequences. The degree of embryonic fragmentation was evaluated by normal morphological assessment prior to transfer and correlated to the blastomere sizes. RESULTS: As a result of normal cell cleavage, mean blastomere size decreased significantly from a volume of 0.28 x 10(6) microm(3) at the 2-cell stage to 0.15 x 10(6) microm(3) at the 4-cell stage (P < 0.001). Mean blastomere size decreased significantly (P < 0.001) with increasing degree of embryonic fragmentation, where highly fragmented embryos showed a 43-67% reduction in blastomere volume compared with embryos with no fragmentation. Multinucleated blastomeres were significantly larger than non-multinucleated blastomeres (P < 0.001). On average, multinucleated blastomeres were 51.5, 67.8 and 73.1% larger than their non-multinucleated sibling blastomeres at the 2-, 3- and 4-cell stage, respectively. Furthermore, the average volume of non-multinucleated blastomeres originating from multinucleated embryos was significantly smaller than the average volume of the blastomeres from mononucleated embryos (P < 0.001). CONCLUSIONS: The results of this study show that the average blastomere size is significantly affected by degree of fragmentation and multinuclearity, and that computer-assisted, multilevel analysis of blastomere size may function as a biomarker for embryo quality. PMID- 14747170 TI - Developmental potential of human pronuclear zygotes in relation to their pronuclear orientation. AB - BACKGROUND: Pronuclear zygote morphology has gained much attention recently due to its possible value in predicting implantation and pregnancy. The present study was performed to assess the developmental potential of zygotes with four different pronuclear orientations. METHODS: This prospective study involved 150 IVF and 190 ICSI patients seeking fertility treatment. Pronuclear zygotes were classified for orientation of the pronuclei in relation to the second polar body placed at the 6 o'clock position. Four types of pronuclear (PN) zygote were recognized, namely PN(1), PN(2), PN(3) and PN(4). The main outcome measures were early cleavage rate, quality of embryos, and implantation and pregnancy rates. RESULTS: The most common types of pronuclei orientations were PN(1) (31.5%) and PN(2) (29.3%), followed by PN(3) (20.5%) and PN(4) (18.5%). A significantly higher proportion of zygotes with PN(1) and PN(4) types of pronuclear orientation underwent early cleavage and developed into grade I embryos compared with other types (P < 0.0001). There was a tendency for higher implantation and pregnancy rates among patients who received embryos developed from PN(1)- and PN(4)-type oocytes, but this was not statistically significant. CONCLUSIONS: Zygotes exhibit four types of pronuclear orientation, and this is independent of the fertilizing spermatozoon or its entry point into the oocytes, whether IVF or ICSI is employed. Early cleavage was associated with PN(1)- and PN(4)-type zygotes, but implantation and pregnancy rates were not associated with pronuclear orientation. Implantation and pregnancy rates tended to be higher for embryos developed from PN(1) and PN(4) pronuclear zygotes. Further studies on a combination of pronuclear orientation and equality of nucleoli may provide a better guide to the implantation potential of embryos. PMID- 14747171 TI - Experimental vitrification of human compacted morulae and early blastocysts using fine diameter plastic micropipettes. AB - BACKGROUND: Vitrification of human blastocysts has been successfully applied using grids, straws and cryoloops. We assessed the survival rate of human compacted morulae and early blastocysts vitrified in pipette tips with a smaller inner diameter and solution volume than the previously described open pulled straw (OPS) method. METHODS: Excess day 5 human embryos (n = 63) were experimentally vitrified in vessels. Embryos were incubated at 37 degrees C with sperm preparation medium (SPM) for 1 min, SPM + 7.5% ethylene glycol (EG)/dimethylsulphoxide (DMSO) for 3 min, and SPM + 16.5% EG + 16.5% DMSO + 0.67 mol/l sucrose for 25 s. They were then aspirated (0.5 microl) into a plastic micropipette tip (0.36 mm inner diameter), exposed to liquid nitrogen (LN(2)) vapour for 2 min before being placed into a pre-cooled cryotube, which was then closed and plunged into LN(2). Embryos were warmed and diluted using 0.33 mol/l and 0.2 mol/l sucrose. RESULTS: The survival rate for compacted morulae was 73% (22/30) and 82% (27/33) for early blastocysts. CONCLUSIONS: The survival rates of human compacted morulae and early blastocysts after vitrification with this simple technique are similar to those reported in the literature achieved by slow cooling and other vitrification protocols. PMID- 14747172 TI - Human cumulus-free oocyte maturational profile and in vitro developmental potential after stimulation with recombinant versus urinary FSH. AB - BACKGROUND: This study compares the influence of recombinant (r)FSH and urinary (u)FSH stimulation on oocyte and embryo quality in patients undergoing ICSI. METHODS AND RESULTS: Denuded oocyte maturity in ICSI patients was graded on a scale from metaphase II (MII) to prophase for nuclear maturation of oocytes. The relationships of cumulus-free oocyte maturational profiles with in vitro outcome parameters were evaluated. In the study population with an unknown distribution of FSH receptor polymorphisms, the ovarian response to rFSH stimulation was significantly different from that of uFSH stimulation, including lower number of oocytes retrieved/oocytes in MII, higher fertilization rates and higher good quality embryo rates. In the study population with a similar distribution of FSH receptor polymorphisms, the ovarian responses to rFSH were lower numbers of oocytes in MII, higher fertilization rates and higher good quality embryo rates, but the total number of oocytes retrieved was not influenced, in comparison with ovarian stimulation with uFSH. CONCLUSIONS: rFSH stimulation appears to influence oocyte quality and subsequent embryo quality in comparison with uFSH stimulation. FSH receptor polymorphisms seem to be an intrinsic factor influencing the ovarian response to FSH stimulation. PMID- 14747173 TI - Anaphase lagging mainly explains chromosomal mosaicism in human preimplantation embryos. AB - BACKGROUND: Cleavage stage embryos as well as postimplantation embryos have been studied extensively over the years. However, our knowledge with respect to the chromosomal constitution of human embryos at the blastocyst stage is still rudimentary. METHODS: In the present paper, a large series of human blastocysts was examined by means of fluorescent in situ hybridization (FISH). RESULTS: It was found that only one in four blastocysts (25%) displayed a normal chromosomal pattern. We defined a group of blastocysts (26%) displaying a simple mosaic chromosome pattern (different cell lines resulting from one chromosomal error), an about equally large group of blastocysts (31%) displaying a complex mosaic chromosome pattern, and a smaller group of blastocysts (11%) showing a chaotic chromosome distribution pattern. Six per cent of all blastocysts analysed could not be assigned one of the previously mentioned chromosomal patterns. CONCLUSION: Anaphase lagging appeared to be the major mechanism through which human embryos acquire a mosaic chromosome pattern during preimplantation development to the blastocyst stage. PMID- 14747174 TI - Quintuplet pregnancy following transfer of two blastocysts: Case report. AB - A 36-year-old single woman presented at the out-patient clinic in March 2000 requesting donor insemination. Between May 2000 and May 2001 she underwent six cycles of intrauterine insemination with donor sperm after clomiphene citrate stimulation without achieving a pregnancy. In January 2002, ICSI was performed; two embryos were transferred on day 3 and a dizygotic bichorionic pregnancy was achieved, which ended in a miscarriage at 21 weeks of gestation. After a second unsuccessful ICSI attempt in which a single embryo transfer was performed, she embarked upon her third attempt in March 2003 at 39 years of age. Two blastocysts were transferred after ICSI, resulting in a quintuplet gestation consisting of a monochorionic biamniotic pregnancy and a monochorionic triamniotic pregnancy. The current case report indicates that monozygotic pregnancies consisting of both twins and triplets are possible after treatment by assisted reproductive technologies. An association between extended culture, manipulation of the zona pellucida, ovarian stimulation and occurrence of monozygotic pregnancies has been suggested by retrospective studies. However, in order to identify more reliably predictive factors for the occurrence of monozygotic pregnancies, it is necessary to perform prospective trials. PMID- 14747175 TI - Twin delivery following 12 years of human embryo cryopreservation: case report. AB - It is uncertain how long IVF units can keep frozen embryos. Few data exist on success of embryo transfer for embryos that have been cryopreserved for many years. We report the delivery of healthy twins following the transfer of embryos cryopreserved for 12 years. To the best of our knowledge, this is the longest reported successful human embryo freezing. PMID- 14747176 TI - Quantifying the changes in endometrial vascularity throughout the normal menstrual cycle with three-dimensional power Doppler angiography. AB - BACKGROUND: We used three-dimensional power Doppler angiography (3D-PDA) to examine the periodic changes in endometrial and subendometrial vascularity during the normal menstrual cycle in 27 women without obvious menstrual dysfunction or subfertility. METHODS: 3D-PDA was performed on alternate days from day 3 of the cycle until ovulation and then every 4 days until menses. Virtual organ computer aided analysis and shell-imaging were used to define and to quantify the power Doppler signal within the endometrial and subendometrial regions producing indices of their relative vascularity. RESULTS: Both the endometrial and subendometrial vascularization index (VI) and vascularization flow index (VFI) increased during the proliferative phase, peaking approximately 3 days prior to ovulation (P < 0.001) before decreasing to a nadir 5 days post-ovulation (P < 0.001). Thereafter, both vascular indices gradually increased during the transition from early to mid-secretory phase. The flow index (FI) showed a similar pattern but with a longer nadir post-ovulation. Smoking was associated with a significantly lower VI and VFI. The FI was significantly lower in women aged > or = 31 years and significantly higher in parous patients. CONCLUSIONS: Endometrial vascularity, as assessed by 3D-PDA, varies significantly during the menstrual cycle and is characterized by a pre-ovulatory peak and post-ovulatory nadir during the peri-implantation window. PMID- 14747177 TI - Menses, fertility and pregnancy after arterial embolization for the control of postpartum haemorrhage. AB - BACKGROUND: The long-term effects of uterine artery embolization for the control of postpartum haemorrhage on menses, fertility and future pregnancy evolution have not been assessed. METHODS: Between November 1993 and July 1999, 31 women with obstetric haemorrhage underwent arterial embolization. Four patients underwent a hysterectomy. Gynaecological information on 25 of the 27 patients who did not undergo hysterectomy was obtained by interview. RESULTS: All women had a return of normal menses. Nine of the 25 patients desired subsequent pregnancy and five patients became pregnant with normal delay of conception. Moreover, two other patients who did not plan another pregnancy became pregnant. A total of 10 pregnancies was studied, four ended during the first trimester. For the six others, the maternal evolution of the pregnancy was uneventful until term. No case of pre-eclampsia was observed. The ultrasonographic examinations revealed normal fetal growth and umbilical and uterine Doppler studies showed no anomaly. No repetition of obstetric haemorrhage was observed. All full-term, newborns were healthy, weighing from 3220 to 4100 g. CONCLUSION: Our results suggest that women who undergo arterial embolization for obstetric haemorrhage should expect to have a return of normal menses with preservation of future fertility and successful uneventful pregnancies. PMID- 14747178 TI - Is previous use of hormonal contraception associated with a detrimental effect on subsequent fecundity? AB - BACKGROUND: The effects of contraception on subsequent fecundity are yet to be substantiated. METHODS: A total of 2841 consecutive pregnant women in Hull and Sheffield completed questionnaires inquiring about time to pregnancy (TTP), contraceptive use, pregnancy planning, previous pregnancies, age and lifestyle characteristics of each partner. Outcome measures were mean TTP, conception probability and odds of subfecundity after discontinuing each contraceptive method. RESULTS: TTP following long-term combined oral contraceptive (COC), short term intrauterine device (IUD) or any duration of injectable use were 2.0-, 1.6-, 3.0-fold longer than TTP after condom use, respectively. Within 6 months of discontinuing COC or injectable use, conception probabilities were 0.86 and 0.34, respectively, whereas those relevant to other methods were not significantly different. All levonorgestrel intrauterine system (IUS) users conceived within 1 month. Relative to condoms, odds of subfecundity after COC, injectable and short term IUD use were 1.9, 5.5, 2.9, respectively. The effect of COC and injectables was stronger with long-term use, in older, obese or oligomenorrhoeic women. Similar results were obtained after adjustment for potential confounders. CONCLUSIONS: A significant reduction in fecundity occurs after COC, IUD or injectables, which is dependent on the duration of use. The effect of COC and injectables is evident in women with a potentially compromised ovarian function. Use of progesterone-only pills or IUS is not associated with a significant effect. PMID- 14747179 TI - High endometrial aromatase P450 mRNA expression is associated with poor IVF outcome. AB - BACKGROUND: The success of IVF treatment is dependent upon embryo quality and coordinated growth and differentiation of the endometrium. Aromatase P450 expression in the human endometrium is thought to be restricted to women with proliferative reproductive tract disorders such as endometriosis, leiomyomas and adenomyosis. METHODS: To determine whether endometrial aromatase P450 mRNA expression is prognostic of IVF outcome, we quantified transcript levels in biopsy specimens from a cohort of subfertile patients awaiting IVF treatment using real-time quantitative PCR. RESULTS: Aromatase P450 transcripts were detected in all endometria examined, although the levels varied considerably between samples, ranging from 0.22 to 486.6 arbitrary units (a.u.). The clinical pregnancy rate in women with high endometrial aromatase P450 mRNA levels (> or = 8.3 a.u.; n = 21) was 9.5% compared with 30.1% in those patients with low expression levels (<8.3 a.u.; n = 101) (P < 0.05). The cycle day of the endometrial biopsy, cause of infertility, age, parity, number of oocytes collected and number of embryos transferred did not differ between patients with high versus low endometrial aromatase P450 mRNA levels (P > 0.1). CONCLUSIONS: Our results indicate that endometrial P450 mRNA levels can identify women at increased risk of IVF treatment failure. PMID- 14747180 TI - C-reactive protein levels in patients undergoing controlled ovarian hyperstimulation for IVF cycle. AB - BACKGROUND: The aim of the present study was to determine serum and follicular fluid C-reactive protein (CRP) levels in patients undergoing controlled ovarian hyperstimulation (COH) for IVF-embryo transfer cycle, and their possible correlation to COH variables. PATIENTS AND METHODS: The subjects were 16 consecutive patients undergoing our routine IVF long GnRH agonist protocol. Blood was drawn three times during the COH cycle: (i) the day on which adequate suppression was obtained (Day-S); (ii) the day of, or prior to HCG administration (Day-HCG); and (iii) the day of (and before) oocyte pick-up (Day-OPU). Levels of sex steroids and serum and follicular fluid CRP were compared among the three time points. Serum and follicular fluid CRP were measured with a commercial immunoturbidimetric assay. RESULTS: Serum levels of CRP were significantly higher on Day-OPU and Day-HCG than on Day-S, and significantly higher on Day-OPU than on Day-HCG. No difference was observed between follicular and serum CRP levels on Day-OPU. No significant correlations were found between serum and follicular fluid CRP, or between serum CRP-to-BMI ratio and serum sex steroid levels or IVF treatment variables. CONCLUSIONS: The significant increase in serum CRP levels during COH, especially after HCG administration, suggests that COH potentiates a state of systemic inflammation. PMID- 14747181 TI - Survival rates during the first trimester of multiple gestations achieved by ICSI: a report of 1448 consecutive multiples. AB - BACKGROUND: The purpose of this study was to determine the rate of spontaneous gestational sac loss during the first trimester in women achieving multiple pregnancies by ICSI. METHODS: A retrospective analysis was performed of 1448 consecutive multiple pregnancies conceived by ICSI. RESULTS: Of the cohort of 1448 pregnancies, twin gestations constituted 59.6% (864), triplets 30.2% (438) and quadruplets 10.0% (146). During the first trimester, 69 (4.7%) patients miscarried, while 179 (12.3%) continued their pregnancies and had fewer gestational sacs at the end of the first trimester than at the beginning. The overall loss rate of any gestational sac during the first trimester in these multiple pregnancies was 10.1%. There was a significant difference in the frequency of spontaneous reduction to twin or singleton pregnancies in the first trimester between women carrying triplets (11.7%) and those carrying quadruplets (3.5%) [P = 0.004; odds ratio (OR) 3.5; 95% confidence interval (CI) 1.3-9.1]. The frequency of gestational sac loss was significantly greater among women >35 years old (20.9%) than in women less than 35 years old (15.9%) (P = 0.03; OR 1.4; 95% CI 1.0-1.9). CONCLUSION: In multiple pregnancies there is a significant risk of spontaneous loss of any embryo during the first trimester. These findings should be considered prior to any decision about selective embryo reduction. PMID- 14747182 TI - Is there a relationship between treatment for infertility and gestational trophoblastic disease? AB - BACKGROUND: The aim of the study was to record the incidence of treatment for infertility prior to development of gestational trophoblastic disease (GTD). METHODS AND RESULTS: A retrospective analysis was undertaken of 231 consecutive women receiving chemotherapy for persistent GTD at Weston Park Hospital, Sheffield, from 1991 to 2001. Three patients in this group had received treatment for infertility prior to their molar pregnancy. In a control group of 226 patients not requiring treatment for persistent GTD, four had had treatment for infertility just before their molar pregnancy, and in a further control group of 208 'normal' pregnancies, eight patients had had treatment for infertility prior to conception. CONCLUSION: We conclude that we can demonstrate no relationship between infertility treatment and subsequent development of GTD. PMID- 14747183 TI - Increased rates of thrombophilia in women with repeated IVF failures. AB - BACKGROUND: We investigated whether hereditary thrombophilia is more prevalent in women with recurrent IVF-embryo transfer failures. METHODS: This case-control study was conducted in an academic tertiary care hospital and compared 45 women with a history of four or more failed IVF cycles (group A) with 44 apparently healthy women matched for age and ethnic origin (group B). All participants were tested for inherited thrombophilias: mutations of prothrombin, factor V Leiden and methylene tetrahydrofolate reductase (MTHFR), and protein C, protein S and antithrombin III deficiencies. RESULTS: Excluding homozygotic MTHFR, the incidence of thrombophilia in group A, was 26.7% compared with 9.1% in group B (P = 0.003; odds ratio 2.9; 95% confidence interval 1.02-8.4). The incidence of thrombophilia in women with unexplained infertility in group A was 42.9% (9/21), compared with 18.2% in group B (P < 0.002). CONCLUSIONS: These data suggest that inherited thrombophilia may play a role in the aetiology of repeated IVF failures, particularly in the subgroup with unexplained fertility. PMID- 14747184 TI - The Polycystic Ovary Syndrome Health-Related Quality of Life Questionnaire (PCOSQ): a validation. AB - BACKGROUND: We wished to evaluate the psychometric properties of the Polycystic Ovary Syndrome Questionnaire (PCOSQ), a questionnaire developed to measure the health-related quality of life (HRQoL) of women with polycystic ovary syndrome. METHOD: To assess reliability and validity, women recruited from an outpatient gynaecology clinic at the Jessop Wing, Royal Hallamshire Hospital, Sheffield completed two copies of the PCOSQ and the Short Form-36 (SF-36). Secondary factor analysis was carried out to verify the composition of the dimensions. Semi structured interviews were conducted to assess face validity. RESULTS: Of the 92 women who consented, 82 women (89%) returned questionnaires at time 1, and 69 women (75%) returned questionnaires at time 2. All five PCOSQ dimensions were internally reliable with Cronbach's alpha scores ranging from 0.70 to 0.97. Intra class correlation coefficients to evaluate test-retest reliability were high (range 0.89-0.95, P < 0.001). Construct validity was demonstrated by high correlations for all comparisons of similar scales of the SF-36 and PCOSQ (0.49 and 0.54). Acne was identified as an important area of HRQoL missing from the questionnaire. CONCLUSIONS: The PCOSQ is a reliable instrument for measuring the HRQoL in women with PCOS. However, the validity of the questionnaire needs to be improved by incorporating a dimension on acne into the instrument. PMID- 14747185 TI - A randomized controlled trial of hysterectomy or levonorgestrel-releasing intrauterine system in the treatment of menorrhagia-effect on FSH levels and menopausal symptoms. AB - BACKGROUND: The purpose of this study was to compare the effects of hysterectomy and a levonorgestrel-releasing intrauterine system (LNG-IUS) on serum FSH levels and menopausal symptoms. METHODS: A total of 236 women referred for menorrhagia to five university hospitals were randomly assigned to treatment with hysterectomy (n = 117) or LNG-IUS (n = 119). Menopausal symptoms were characterized by the Kupperman menopausal distress test. Serum FSH and estradiol levels were measured at baseline and 6 and 12 months after hysterectomy or application of LNG-IUS. Analyses were by intention to treat. RESULTS: After 6 months, there was no difference between the groups, but 12 months after follow-up hysterectomized women had higher FSH levels than women with LNG-IUS (P = 0.005). There was a significant association between FSH levels and treatment modality (P = 0.020). Hot flushes increased significantly in the hysterectomy group (P = 0.02). There was a significant association between hot flushes and both treatment modality and age (P = 0.02 and P = 0.01, respectively). CONCLUSION: Hysterectomy may impair ovarian function shown by rising serum FSH levels and hot flushes. However, these results should be interpreted with caution, and longer follow-up is needed. PMID- 14747186 TI - Predictive value of menstrual cycle pattern, body mass index, hormone levels and polycystic ovaries at age 15 years for oligo-amenorrhoea at age 18 years. AB - BACKGROUND: On the question of how to counsel adolescents with irregular menstrual cycles or oligomenorrhoea, no clear answer has been given. Adolescents with oligomenorrhoea especially show endocrine abnormalities and may be at risk for ovulatory dysfunction and the polycystic ovary syndrome in adulthood. METHODS: We followed a cohort of adolescents to document changes in menstrual cycle pattern between ages 15 and 18 years in the general population. RESULTS: Two per cent (2/128) of adolescents with regular menstrual cycles developed oligomenorrhoea, and 12% (17/148) of those with irregular menstrual cycles did so. Fifty-one per cent (34/67) of the oligomenorrhoeic adolescents remained oligomenorrhoeic. Increase in body mass index (BMI), concentration of LH, androstenedione or testosterone, and polycystic ovaries (PCO) were associated with persistence of oligomenorrhoea. In multivariate analysis only a normal to high BMI (>19.6 kg/m(2)) consistently contributed significantly to predict persistent oligomenorrhoea. Glucose:insulin ratio as a marker for insulin resistance was not associated with an increased risk for oligomenorrhoea. CONCLUSIONS: Oligomenorrhoea at age 18 years is better predicted by menstrual cycle pattern at age 15 years than by LH or androgen concentrations or PCO at this age. Not only obese, but also normal weight oligomenorrhoeic, adolescents have a high risk of remaining oligomenorrhoeic. PMID- 14747187 TI - Analysis of cyclooxygenase-2 (COX-2) expression in different sites of endometriosis and correlation with clinico-pathological parameters. AB - BACKGROUND: Recent studies have demonstrated the overexpression of cyclooxygenase 2 (COX-2) in endometriosis. The aim of this study was to investigate the expression of COX-2 in different anatomical sites of endometriosis and its association with clinico-pathological parameters in a single institutional series of patients undergoing operative treatment. METHODS: COX-2 expression was analysed by immunohistochemistry in 136 samples of endometriotic tissue from 103 patients affected by endometriosis. RESULTS: COX-2 immunoreaction was observed in 78.5% of ovarian endometriotic cysts, in 11.1% of peritoneal implants and 13.3% of recto-vaginal nodules. COX-2 positivity was not distributed differently according to age, pre-operative serum levels of CA125 and AFS score. Moreover, COX-2 positivity did not show any significant variation according to the subjective intensity of pain, as dysmenorrhoea, chronic pelvic pain, lower urinary tract or gastrointestinal symptoms, or according to infertility. CONCLUSIONS: Increased COX-2 expression in the endometriotic ovarian cyst wall was observed with respect to other extraovarian localizations. No relevant correlations between COX-2 positivity and clinico-pathological characteristics and symptoms of patients were observed. PMID- 14747188 TI - Differential FSH exposure in preantral follicle culture has marked effects on folliculogenesis and oocyte developmental competence. AB - BACKGROUND: We investigated at what stage early cultured preantral mouse follicles become dependent on a minimal effective FSH dose (10 mIU/ml) and analysed the influence of implementing FSH at several time-points during in vitro culture. METHODS: Two-layered mouse follicles were cultured for 12 days under seven different FSH exposure regimens and ovulated on day 13 by hCG/EGF. Ovulated cumulus-oocyte complexes were fertilized and embryos were cultured up to the blastocyst stage. RESULTS: When FSH was absent or added only once at the start of culture, follicle survival was significantly reduced (22 and 52% respectively versus 95% when FSH was continuously present, P < 0.01) and oocyte quality was compromised, providing few oocytes for embryo culture (19 and 7% versus 71% in continuous presence of FSH, P < 0.01). Optimal follicle survival rates can be ensured by implementing FSH at the latest from day 4 of culture. By introducing FSH later than day 4, follicle survival rates and number of ovulated oocytes decreased. Estradiol production and luteinization were strongly related to the moment of introducing FSH in culture. Fertilization and preimplantation embryo development rate were found to be highest in cultures where FSH support was implemented during the preantral stage. CONCLUSION: Exposure to FSH before formation of the antral-like cavity had a positive effect on follicle survival and oocyte robustness. PMID- 14747189 TI - Expression of fractalkine in the Fallopian tube and of CX3CR1 in sperm. AB - BACKGROUND: Fractalkine is a CX(3)C chemokine that has chemoattractant activity for T cells, monocytes and natural killer (NK) cells. The objective of this study was 2-fold: to evaluate (i) the presence of fractalkine in the Fallopian tube and (ii) the existence of CX(3)CR1 (fractalkine receptor) in ejaculated sperm. METHODS AND RESULTS: Western blot analysis revealed that fractalkine protein was detected as a 95 kDa band in the isthmus, the ampulla and the infundibulum of the Fallopian tube. Immunohistochemistry revealed positive staining of epithelial cells in the Fallopian tube. RT-PCR demonstrated that fractalkine transcripts were expressed in all parts of the Fallopian tube. RT-PCR also revealed that CX(3)CR1-positive cells were present in the Fallopian tube. CX(3)CR1-positive cells were present in the stroma of the Fallopian tube. The villi of the ciliated cells were positively stained. To determine the function of fractalkine in the Fallopian tube, we examined whether CX(3)CR1 was present in ejaculated sperm. RT PCR demonstrated that CX(3)CR1 transcripts were expressed in the ejaculated sperm. Immunohistochemistry demonstrated positive staining of the tail of the spermatozoa. CONCLUSIONS: The present findings suggest that fractalkine in the Fallopian tube contributes to the immunodefence mechanism during fertilization and to the sperm motion in the oviduct. PMID- 14747190 TI - Lipid, glucose and homocysteine metabolism in women treated with a GnRH agonist with or without raloxifene. AB - BACKGROUND: Although GnRH analogues are widely used to treat a variety of sex hormone-related diseases, little is known about their effect on metabolism. Therefore, we have evaluated the effect of a GnRH analogue, administered with or without raloxifene, on serum levels of lipoproteins, glucose, insulin and homocysteine (Hcy). METHODS: One hundred premenopausal women with symptomatic uterine leiomyomas were initially enrolled and randomized to receive 3.75 mg/28 days leuprolide acetate depot associated with 60 mg/day raloxifene hydrochloride (group A) or 1 placebo tablet/day (group B) for six cycles of 28 days. At entry and at cycle 6, subjects underwent anthropometric measurements, including body mass index and waist-to-hip ratio measurements, and blood chemistry assays for serum total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C), triglycerides (TG), glucose, insulin, Hcy, vitamin B(12) and folate concentrations. Insulin resistance was evaluated with the homeostasis model assessment (HOMA) score. RESULTS: Baseline parameters were similar in the two groups. At cycle 6, TC, HDL-C, LDL-C and TG levels were significantly increased (P < 0.05) in group B. In group A, LDL-C levels were unchanged, and TC, HDL-C and TG levels were increased (P < 0.05). Serum TC and LDL-C levels differed (P < 0.05) between the groups. Glucose levels were unchanged between and within groups, whereas insulin levels and HOMA scores increased (P < 0.05) versus baseline in group B. Post-treatment Hcy levels were higher (P < 0.05) versus baseline in group B; they were unchanged in group A. Serum vitamin B(12) and folate concentrations were unchanged in both groups. CONCLUSIONS: GnRH analogues alter serum lipoprotein and Hcy levels and increase insulin resistance. These acute metabolic changes may be prevented or reduced by raloxifene. PMID- 14747191 TI - Effects of breastfeeding chemosignals on the human menstrual cycle. AB - BACKGROUND: To date, there has not been an investigation to determine whether lactating women and their infants influence the ovarian function of other women with whom they interact. METHODS: In a randomized, double-blind, placebo controlled study of 47 nulliparous women, we utilized both within- and between subjects controls to assess the effects of sustained exposure to breastfeeding compounds on menstrual cycle length, as well as characteristics of each phase of the ovarian cycle. RESULTS: Breastfeeding compounds modulated ovarian cycle length in comparison with the carrier control (0.01 < or = all P values < or = 0.05), disrupting the normal homeostatic regulation of cycle length and tripling its variance. Hence, women with long cycles stayed long and did not regress to the mode of 29 days and women with short cycles maintained short cycles. This effect was driven by changes in both the follicular and luteal phases of the cycle (0.01 < or = all P values < or = 0.04) and changed the timing of the pre ovulatory surge of LH. CONCLUSIONS: Because compounds from lactating women and their infants modulated the ovarian cycles of women, as is seen in other mammals, they have the potential to function as pheromones, regulating fertility within groups of women. PMID- 14747192 TI - Basal FSH concentrations as a marker of ovarian ageing are not related to pregnancy outcome in a general population of women over 30 years. AB - BACKGROUND: Previous studies suggest that elevated basal FSH concentrations are related to aneuploid pregnancies. However, there have been no prospective studies evaluating the incidence of aneuploidies in relation to basal FSH concentrations. Since the majority of aneuploid conceptions end in early pregnancy loss or abortion of a recognized pregnancy, these determinants are appropriate intermediate end-points to study aneuploidy. METHODS: We performed a prospective study in 129 women without a history of subfertility pursuing a spontaneous pregnancy. Basal FSH concentrations were measured during three menstrual cycles. Urinary HCG levels were measured during menstruation for a maximum of six menstrual cycles, to detect early pregnancy loss. We estimated the effect of basal FSH concentrations on pregnancy outcome, taking into account possible confounders. RESULTS: We observed no significant effect of basal FSH concentrations on the incidence of early pregnancy loss or abortion of clinically recognized pregnancies. CONCLUSIONS: We conclude that in a population of women without a history of subfertility, pursuing a spontaneous pregnancy, basal FSH concentrations are not related to the incidence of early pregnancy loss or abortions. This prospective study therefore fails to confirm a relationship between signs of decreased ovarian reserve and aneuploid pregnancies. PMID- 14747193 TI - Neonatal outcome in a Danish national cohort of 3438 IVF/ICSI and 10,362 non IVF/ICSI twins born between 1995 and 2000. AB - BACKGROUND: In Denmark, one-third of twin pregnancies are the result of IVF/ICSI treatment. Limited data on neonatal outcome in IVF/ICSI twins are available in the literature. METHODS: A register study was conducted on neonatal morbidity and mortality in a complete national twin cohort including all 3438 (3393 live-born) IVF/ICSI and 10,362 (10,239 live-born) non-IVF/ICSI twins born between 1995 and 2000. Twins were identified in the National Medical Birth Registry and dichotomized into IVF/ICSI and non-IVF/ICSI by cross-reference with the Danish IVF Registry. Data on neonatal morbidity and mortality were retrieved from the Danish Patient Registry and the Danish Registry of Causes of Deaths. In order to exclude monozygotic twins, sub-analyses on unlike-sex twins were conducted. RESULTS: A birth weight discordance of >20% was observed in 20.6% of IVF/ICSI versus 15.7% of control twin pairs (P < 0.001). The risk of discordant birth weight >20% was OR 1.29 (95% CI 1.04-1.58) in unlike-sex IVF/ICSI twins versus control twins. The risk of delivery at <37 completed weeks and birth weight <2500 g was similar in the two cohorts; however, in unlike-sex IVF/ICSI versus control twins the risk of delivery at <37 weeks and birth weight <2500 g was OR 1.22 (95% CI 1.09-1.38) and OR 1.25 (1.11-1.40) respectively. After stratification for maternal age and parity, these risks disappeared. IVF/ICSI twins carried a higher risk of admittance to a neonatal intensive care unit (NICU) than control twins (OR 1.18, 95% CI 1.09-1.27), and this was even more pronounced in unlike-sex twins [OR 1.34 (95% CI 1.19-1.51)]. No differences were observed in malformation or mortality rates between the two cohorts. CONCLUSIONS: Despite higher birth weight discordance and more NICU admissions among IVF/ICSI twins, neonatal outcome in IVF/ICSI twins seems to be comparable with that of non-IVF/ICSI twins, when only dizygotic twins were considered in the comparisons. PMID- 14747194 TI - Influences of maternal weight on the secondary sex ratio of human offspring. AB - BACKGROUND: Less than optimal reproductive conditions may be associated with a secondary sex ratio biased towards females. Body weight represents a critical determinant of reproduction. Accordingly, we evaluated whether preconception body weight and weight gain during pregnancy influence the sex ratio of human offspring. METHODS: A retrospective study was performed on 10 239 births in 1997 2001. Time of conception, preconception body weight and net weight gain during pregnancy were obtained for 9284 pregnancies. RESULTS: The secondary sex ratio of mothers in the lowest quartile of pre-pregnancy body weight (<54.6 kg) was lower than that of the other three quartiles (0.497 versus 0.525; P < 0.01). In contrast, the sex ratio of children born by the women in the highest quartile of weight gain during pregnancy appeared lower than that of the first three quartiles (0.493 versus 0.516; P = 0.054). CONCLUSIONS: A low pre-pregnancy weight and a greater weight gain during pregnancy are both associated with a reduced secondary sex ratio. These data indicate that in women with non-optimal reproductive/metabolic conditions, a greater attrition is exerted on male than on female offspring. PMID- 14747195 TI - Effects of season of birth on reproduction in contemporary humans: brief communication. AB - BACKGROUND: At high latitudes the external environment varies with season, and therefore the season of birth may contribute to the developmental processes during the perinatal period. METHODS: We investigated the association between birth season and measures of reproductive performance (offspring count, percentage childless individuals) in a contemporary sample of women and men. RESULTS: In the male sample (n = 2342), men born in autumn had fewer offspring (mean 1.4 versus 1.62; P < 0.01) and a higher probability of remaining childless (32.6% versus 25.6%; P = 0.01) than men born in spring. The photoperiod at a male's birth was significantly positively correlated with his subsequent offspring count (P = 0.023). In the female sample, an association between birth season and reproduction was not found. CONCLUSIONS: We assume that in men, among other seasonal factors, pre- or perinatal photoperiod might be involved in the underlying physiological mechanism. PMID- 14747196 TI - Familial aggregation of endometriosis in a large pedigree of rhesus macaques. AB - BACKGROUND: Endometriosis occurs in several non-human primate species that have menstrual cycles. This study investigated the prevalence and familial aggregation of endometriosis in one of those species, the rhesus macaque. METHODS: Between 1978 and 2001, 142 animals with endometriosis were identified from necropsy and surgical records and through the use of magnetic resonance imaging (MRI) at the Wisconsin National Primate Research Center, Madison, USA. All cases were used to build one large multigenerational pedigree and nine nuclear families comprising 1602 females in total. By 2002, the pedigrees contained 124 cases diagnosed at necropsy; 17 at surgery and three at MRI. Female animals that had died aged > or = 10 years without endometriosis, had both ovaries until at least 1 year prior to death, and had a full necropsy, were considered unaffected. RESULTS: The prevalence of endometriosis among necropsied animals aged > or = 10 years in the colony was 31.4% [95% confidence interval (CI) 26.9-35.9%]; prevalence increased with rising age and calendar age at death. Familial aggregation of endometriosis was strongly suggested by a significantly higher average kinship coefficient among affecteds compared with unaffecteds (P < 0.001) and a higher recurrence risk for full sibs (0.75; 95% CI 0.45-1.0) compared with maternal half sibs (0.26; 95% CI 0.10-0.41) and paternal half sibs (0.18; 95% CI 0.02-0.34). The segregation ratio among affected mothers (44.2%) was not significantly higher compared with unaffected mothers (36.6%). CONCLUSIONS: The results support familial aggregation of endometriosis in the rhesus macaque, and indicate that this is a promising animal model for the investigation of mode of inheritance, the location of potential genetic susceptibility loci and the influence of environmental factors. PMID- 14747197 TI - Deficiency of 17,20-lyase causing giant ovarian cysts in a girl and a female phenotype in her 46,XY sister: case report. AB - A 13-year-old girl was referred because of progressive abdominal pain caused by ovarian torsion and giant ovarian cysts. Secondary sexual characteristics were absent. Hormone analysis revealed markedly elevated serum levels of progesterone and 17-hydroxyprogesterone in combination with very low peripheral concentrations of C19 steroids (dehydroepiandrosterone and androstenedione) and estrogens. Serum concentrations of FSH and LH exceeded the upper limit of normal levels in adult women. The patient's 16-year-old 46,XY sibling showed a female phenotype with similar hormonal disturbances. Both siblings were found to be compound heterozygotes for two mutations in the CYP17 gene: an R347C mutation in one allele and a 25-base pair deletion in exon 1 in the other. The resulting block in 17,20-lyase activity caused an inability to synthesize androgens and estrogens, and increased levels of gonadotrophins due to a lack of negative feedback. The increased levels of gonadotrophins most likely stimulated growth of the ovarian cysts. The administration of a GnRH antagonist reduced the size of the cysts within a few weeks. At present, the girl is being treated with a combination of a GnRH agonist and hormone replacement therapy. PMID- 14747198 TI - Taskforce 7: Ethical considerations for the cryopreservation of gametes and reproductive tissues for self use. AB - This seventh statement of the ESHRE Task Force on Ethics and Law considers ethical questions and specific dilemmas surrounding the cryopreservation of gametes and reproductive tissue. This is of particular relevance with advancing techniques for cryopreservation and the desire of many individuals to preserve fertility after cancer, chronic illness, iatrogenic complications of treatment or simply with advancing age. PMID- 14747199 TI - What is the LH ceiling level for follicular growth arrest in late follicular phase? PMID- 14747201 TI - Sex ratio of birth during wartime and psychological tensions. PMID- 14747203 TI - A randomized trial evaluating a predominantly fetal growth-based strategy to guide management of gestational diabetes in Caucasian women. AB - OBJECTIVE: To compare the management of Caucasian women with gestational diabetes (GDM) based predominantly on monthly fetal growth ultrasound examinations with an approach based solely on maternal glycemia. RESEARCH DESIGN AND METHODS: Women with GDM who attained fasting capillary glucose (FCG) <120 mg/dl and 2-h postprandial capillary glucose (2h-CG) <200 mg/dl after 1 week of diet were randomized to management based on maternal glycemia alone (standard) or glycemia plus ultrasound. In the standard group, insulin was initiated if FCG was repeatedly >90 mg/dl or 2h-CG was >120 mg/dl. In the ultrasound group, thresholds were 120 and 200 mg/dl, respectively, or a fetal abdominal circumference >75th percentile (AC>p75). Outcome criteria were rates of C-section, small-for gestational-age (SGA) or large-for-gestational-age (LGA) infants, neonatal hypoglycemia (<40 mg/dl), and neonatal care admission. RESULTS: Maternal characteristics and fetal AC>p75 (36.0 vs. 38.4%) at entry did not differ between the standard (n = 100) and ultrasound groups (n = 99). Assignment to (30.0 vs. 40.4%) and mean duration of insulin treatment (8.3 vs. 8.1 weeks) did not differ between groups. In the ultrasound group, AC>p75 was the sole indication for insulin. The ultrasound-based strategy, as compared with the maternal glycemia only strategy, resulted in a different treatment assignment in 34% of women. Rates of C-section (19.0 vs. 18.2%), LGA (10.0 vs. 12.1%), SGA (13.0 vs. 12.1%), hypoglycemia (16.0 vs. 17.0%), and admission (15.0 vs. 14.1%) did not differ significantly. CONCLUSIONS: GDM management based on fetal growth combined with high glycemic criteria provides outcomes equivalent to management based on strict glycemic criteria alone. Inclusion of fetal growth might provide the opportunity to reduce glucose testing in low-risk pregnancies. PMID- 14747204 TI - Relation between development of nephropathy and the p22phox C242T and receptor for advanced glycation end product G1704T gene polymorphisms in type 2 diabetic patients. AB - OBJECTIVE: The development of diabetic nephropathy is considered to be associated with oxidative stress. NADPH oxidase and the receptor for advanced glycation end products (RAGE) have attracted attention as mechanisms of generating oxidative stress. We studied the relation between the genotypes of the NADPH p22phox C242T and RAGE G1704T polymorphisms and the development of diabetic nephropathy in type 2 diabetic patients. RESEARCH DESIGN AND METHODS: Using a retrospective review of clinical data, we allocated 181 Japanese type 2 diabetic patients to one of two groups: patients without diabetic nephropathy (group N; n = 108) and patients developing diabetic nephropathy (group D; n = 73) for 10 years or more. The p22phox C242T and RAGE G1704T polymorphisms were examined by Taqman PCR methods. RESULTS: The frequency of the p22phox CC genotype was significantly higher in group D than in group N (90 vs. 79%; P = 0.0427). The frequency of the RAGE GT + TT genotype was significantly higher in group D than in group N (26 vs. 13%; P = 0.0313). The frequency of the combination of p22phox CC and RAGE GT + TT genotypes was significantly higher in group D than in group N (22 vs. 8%; P = 0.0057). In multiple logistic regression analysis, systolic blood pressure, HbA(1c), triglycerides, and the combination of polymorphisms were shown to be independent variables. CONCLUSIONS: These results suggest that assessment of the combination of NADPH p22phox C242T and RAGE G1704T polymorphisms may be useful in identifying the risk for developing diabetic nephropathy in type 2 diabetic patients. PMID- 14747205 TI - Sex-related differences between adiponectin and insulin resistance in schoolchildren. AB - OBJECTIVE: To study the effect of body composition and adiponectin on insulin resistance and beta-cell function in schoolchildren during puberty. RESEARCH DESIGN AND METHODS: Plasma adiponectin level and its relationships with insulin sensitivity and beta-cell function were analyzed in 500 randomly recruited nondiabetic Taiwanese schoolchildren (245 boys and 255 girls) aged 6-18 years in a national survey program for diabetes in 1999. Insulin resistance and beta-cell function were evaluated by homeostasis model assessment (HOMA). Plasma adiponectin concentrations were determined with radioimmunoassay. RESULTS: Plasma glucose levels remained stable, whereas insulin resistance increased with a compensatory rise in beta-cell function during this period. A transient drop of adiponectin level with a trough at 10-12 years was found in boys but not in girls. This pubertal drop of adiponectin levels in boys coincides with the sharp rise in testosterone concentration. A negative correlation between testosterone levels and adiponectin concentration was also noted in boys (r = -0.142, P = 0.032). Plasma adiponectin levels correlated inversely with relative body weight, fasting insulin concentrations, and insulin resistance index by HOMA in boys aged 15-18 years and in girls aged 11-14 years. No association was observed between adiponectin levels and beta-cell function by HOMA. CONCLUSIONS: There is a transient drop in the level of adiponectin during male puberty, correlated with the increase in testosterone level in boys. Plasma adiponectin levels were inversely correlated with obesity and insulin resistance in boys and girls during the pubertal period. PMID- 14747206 TI - Indexes of insulin resistance and secretion in obese children and adolescents: a validation study. AB - OBJECTIVE: To assess the concurrent validity of fasting indexes of insulin sensitivity and secretion in obese prepubertal (Tanner stage 1) children and pubertal (Tanner stages 2-5) adolescents using estimates from the modified minimal model frequently sampled intravenous glucose tolerance test (FSIVGTT) as a criterion measure. RESEARCH DESIGN AND METHODS: Eighteen obese children and adolescents (11 girls and 7 boys, mean age 12.2 +/- 2.4 years, mean BMI 35.4 +/- 6.2 kg/m(2), mean BMI-SDS 3.5 +/- 0.5, 7 prepubertal and 11 pubertal) participated in the study. All participants underwent an insulin-modified FSIVGTT on two occasions, and 15 repeated this test a third time (mean 12.9 and 12.0 weeks apart). S(i) measured by the FSIVGTT was compared with homeostasis model assessment (HOMA) of insulin resistance (HOMA-IR), quantitative insulin sensitivity check index (QUICKI), fasting glucose-to-insulin ratio (FGIR), and fasting insulin (estimates of insulin sensitivity derived from fasting samples). The acute insulin response (AIR) measured by the FSIVGTT was compared with HOMA of percent beta-cell function (HOMA-beta%), FGIR, and fasting insulin (estimates of insulin secretion derived from fasting samples). RESULTS: There was a significant negative correlation between HOMA-IR and S(i) (r = -0.89, r = -0.90, and r = -0.81, P < 0.01) and a significant positive correlation between QUICKI and S(i) (r = 0.89, r = 0.90, and r = 0.81, P < 0.01) at each time point. There was a significant positive correlation between FGIR and S(i) (r = 0.91, r = 0.91, and r = 0.82, P < 0.01) and a significant negative correlation between fasting insulin and S(i) (r = -90, r = -0.90, and r = -0.88, P < 0.01). HOMA-beta% was not as strongly correlated with AIR (r = 0.60, r = 0.54, and r = 0.61, P < 0.05). CONCLUSIONS: HOMA-IR, QUICKI, FGIR, and fasting insulin correlate strongly with S(i) assessed by the FSIVGTT in obese children and adolescents. Correlations between HOMA-beta%, FGIR and fasting insulin, and AIR were not as strong. Indexes derived from fasting samples are a valid tool for assessing insulin sensitivity in prepubertal and pubertal obese children. PMID- 14747207 TI - Low cardiorespiratory fitness is associated with elevated C-reactive protein levels in women with type 2 diabetes. AB - OBJECTIVE: The purpose of this study was to examine differences in novel markers of cardiovascular disease (CVD) in women with type 2 diabetes stratified according to cardiorespiratory fitness. RESEARCH DESIGN AND METHODS: A total of 28 women (mean age 57 +/- 6 years) with type 2 diabetes who were free from overt CVD were placed into low cardiorespiratory fitness (LCF) or average cardiorespiratory fitness (ACF) groups based on a graded exercise test to exhaustion. A group of eight women without type 2 diabetes were also examined and served as healthy control subjects. The median VO(2peak) value was used as a cutoff for group determination. We assessed both conventional CVD risk factors, including blood pressure, BMI, and lipid profile, as well as novel CVD risk factors, such as left ventricular filling dynamics, arterial stiffness, fasting insulin, and C-reactive protein (CRP). RESULTS: VO(2peak) values were 69 +/- 14 and 91 +/- 24% of predicted values for sedentary age-matched healthy individuals in the LCF and ACF groups, respectively. BMI was significantly greater in the LCF group (P < 0.05); however, no differences were observed in age, lipid profile, or resting hemodynamics. CRP was 3.3-fold higher in the LCF group (6.3 +/- 41. vs. 1.9 +/- 1.7 mg/l, P < 0.05), whereas other novel markers of CVD were not significantly different between the groups. Significant negative relationships were observed between VO(2peak) and both CRP (r = -0.49) and the homeostasis model assessment index (r = -0.48) (P < 0.05). CONCLUSIONS: The novel finding of this investigation is that low cardiorespiratory fitness is associated with elevated CRP and reduced fasting glucose control in women with type 2 diabetes. PMID- 14747208 TI - A prospective study of glycemic control during holiday time in type 2 diabetic patients. AB - OBJECTIVE: In the U.K. Prospective Diabetes Study, A1C increased from 1.2 to 1.7% and fasting plasma glucose from 1.0 to 2.8 mmol/l over 10 years in type 2 diabetic patients. It is not known whether the blood glucose increase observed in long-term studies of type 2 diabetes results from small, steady increases throughout the year or from increases during discrete periods. RESEARCH DESIGN AND METHODS: To estimate the variation of actual glycemic control and its relation to holiday times, we measured A1C and fructosamine in 110 patients with type 2 diabetes. These measurements were performed four times at intervals of 4-6 weeks; therefore, glycemic change was determined for three periods: preholiday period (from between November 13 and December 20 to between December 20 and January 20), holiday period (from between December 20 and January 20 to between January 28 and February 28), and postholiday period (from between January 28 and February 28 to between March 1 and April 10). A final measurement of A1C was obtained from 90 subjects in the following December or January. RESULTS: The mean A1C increased, but not significantly, during the preholiday (increase 0.135 +/- 0.723%, P = 0.055) and holiday (increase 0.094 +/- 0.828%, P = 0.239) periods. The mean A1C decreased, but not significantly, during the postholiday period (decrease 0.022 +/- 0.588%, P = 0.695). Altogether, the A1C change during these three periods increased significantly (increase 0.207 +/- 0.943%, P = 0.024). The mean fructosamine increased significantly during the preholiday period (increase 0.151 +/- 0.460 mmol/l, P = 0.001), but there was no significant change during the holiday period (increase 0.057 +/- 0.593 mmol/l, P = 0.321). However, fructosamine decreased significantly during the postholiday period (decrease 0.178 +/- 0.448 mmol/l, P < 0.001). Altogether, the fructosamine changes during the study periods showed no significant difference (increase 0.030 +/- 0.566 mmol/l, P = 0.579). Between March or early April and the following December or January, there was no additional change in A1C (decrease 0.009 +/- 1.039%, P = 0.935) for the 90 participants who returned for follow-up treatment. CONCLUSIONS: The present study demonstrates an influence of winter holidays on the glycemic control of patients who have type 2 diabetes, and this poor glycemic control might not be reversed during the summer and autumn months. Therefore, the cumulative effects of the yearly A1C gain during the winter holidays are likely to contribute to the substantial increase in A1C that occurs every year among type 2 diabetic individuals. PMID- 14747209 TI - Health literacy and pregnancy preparedness in pregestational diabetes. AB - OBJECTIVE: We investigated the association between functional health literacy and markers of pregnancy preparedness in women with pregestational diabetes. RESEARCH DESIGN AND METHODS: English- and Spanish-speaking pregnant women with pregestational diabetes were recruited. Women completed the Test of Functional Health Literacy in Adults (TOFHLA) short form and a questionnaire. A TOFHLA score of < or =30 was defined as low functional health literacy. RESULTS: Of 74 women participating in the study, 16 (22%) were classified as having low functional health literacy. Compared with women with adequate health literacy, those with low health literacy were significantly more likely to have an unplanned pregnancy (P = 0.02) and significantly less likely to have either discussed pregnancy ahead of time with an endocrinologist or obstetrician (P = 0.01) or taken folic acid (P = 0.001). CONCLUSIONS: The results of this study suggest that low functional health literacy among women with pregestational diabetes is associated with several factors that may adversely impact birth outcomes. PMID- 14747210 TI - Utility of casual postprandial glucose levels in type 2 diabetes management. AB - OBJECTIVE: Because readily available glycemic indicators are needed to guide clinical decision-making for intensification of diabetes therapy, our goals were to define the relationship between casual postprandial plasma glucose (cPPG) levels and HbA(1c) in patients with type 2 diabetes and to determine the predictive characteristics of a convenient glucose cutoff. RESEARCH DESIGN AND METHODS: We examined the relationship between cPPG levels (1-4 h post meal) and HbA(1c) levels in 1,827 unique patients who had both determinations during a single office visit. RESULTS: The population studied was predominantly African American and middle-aged, with average cPPG of 201 mg/dl and HbA(1c) of 8.4%. The prevalence of HbA(1c) > or = 7.0% was 67% and HbA(1c) >6.5% was 77%. Overall, cPPG and HbA(1c) were linearly correlated (r = 0.63, P < 0.001). The correlation between cPPG and HbA(1c) was strongest in patients treated with diet alone (n = 348, r = 0.75, P < 0.001) and weaker but still highly significant for patients treated with oral agents (n = 610, r = 0.64, P < 0.001) or insulin (n = 869, r = 0.56, P < 0.001). A cutoff cPPG >150 mg/dl predicted an HbA(1c) level > or = 7.0% in the whole group, with a sensitivity of 78%, a specificity of 62%, and an 80% positive predictive value. The same cPPG cutoff of 150 mg/dl predicted an HbA(1c) level >6.5%, with a sensitivity of 74%, a specificity of 66%, and an 88% positive predictive value. CONCLUSIONS: When rapid-turnaround HbA(1c) determinations are not available, a single cPPG level >150 mg/dl may be used during a clinic visit to identify most inadequately controlled patients and allow timely intensification of therapy. PMID- 14747211 TI - The effect of the PREMIER interventions on insulin sensitivity. AB - OBJECTIVE: This ancillary study of PREMIER sought to determine the effects on insulin sensitivity of a comprehensive behavioral intervention for hypertension with and without the Dietary Approaches to Stop Hypertension (DASH) dietary pattern. RESEARCH DESIGN AND METHODS: Participants were assigned to one of three nonpharmacologic interventions for blood pressure (group A, advice only; group B, established; and group C, established plus DASH). The established intervention included weight loss, reduced sodium intake, increased physical activity, and moderate alcohol intake; the DASH dietary pattern was added to the established intervention for those in group C. The DASH dietary pattern is high in fruits, vegetables, and low-fat dairy products while being lower in total fat, saturated fat, and cholesterol. It is abundant in nutrients such as magnesium, calcium, and protein, which have been associated with improved insulin sensitivity. Insulin sensitivity was measured at baseline and at 6 months using the frequently sampled intravenous glucose tolerance test with minimal model analysis. RESULTS: Both intervention groups decreased total calories, percentage of calories from fat, and sodium intake to similar levels, with similar amounts of energy expenditure and weight loss. Covariate differences seen only in group C included increased intake of protein, potassium, calcium, and magnesium. Compared with control subjects, insulin sensitivity improved significantly only in group C, from 1.96 to 2.95 (P = 0.047). Group B did have a significant decrease in fasting insulin and glucose, but the changes in insulin sensitivity did not reach statistical significance when compared with control subjects. CONCLUSIONS: These results suggest that including the DASH dietary pattern as part of a comprehensive intervention for blood pressure control enhances insulin action beyond the effects of a comprehensive intervention that does not include DASH. PMID- 14747212 TI - Treatment of type 2 diabetes in childhood using a very-low-calorie diet. AB - OBJECTIVE: Pharmacologic agents currently approved for use in children with type 2 diabetes (metformin and insulin) are less than optimal for some patients. We evaluated the use of a ketogenic, very-low-calorie diet (VLCD) in the treatment of type 2 diabetes. RESEARCH DESIGN AND METHODS: We conducted a chart review of 20 children (mean age 14.5 +/- 0.4 years) who consumed a ketogenic VLCD in the treatment of type 2 diabetes. Several response variables (BMI, blood pressure, HbA(1c), blood glucose, and treatment regimens) were examined before, during, and up to 2 years after the diet and compared with a matched diabetic control group. RESULTS: Before starting the diet, 11 of 20 patients were treated with insulin and 6 with metformin. Mean daily blood glucose values fell from 8.9 +/- 1.1 to 5.5 +/- 0.38 mmol/l (P < 0.0001) in the first 3 days of the VLCD, allowing insulin and oral agents to be discontinued in all but one subject. BMI fell from 43.5 +/- 1.8 to 39.3 +/- 1.8 kg/m(2) (P < 0.0001) and HbA(1c) dropped from 8.8 +/ 0.6 to 7.4 +/- 0.6% (P < 0.005) as the diet was continued for a mean of 60 +/- 8 days (range 4-130 days), and none required resumption of antidiabetic medications. Sustained decreases in BMI and insulin requirements were observed in patients remaining on the VLCD for at least 6 weeks when compared with those of the control group. CONCLUSIONS: The ketogenic VLCD is an effective short-term, and possibly long-term, therapy for pediatric patients with type 2 diabetes. Blood glucose control and BMI improve, allowing the discontinuation of exogenous insulin and other antidiabetic agents. This diet, although strict, has potential as an alternative to pharmacologic therapies for this emerging subset of diabetic individuals. PMID- 14747213 TI - Ethnic differences in insulin sensitivity and beta-cell function in premenopausal or early perimenopausal women without diabetes: the Study of Women's Health Across the Nation (SWAN). AB - OBJECTIVE: To assess differences in insulin sensitivity and beta-cell function between nondiabetic premenopausal or early perimenopausal non-Hispanic white women and African American, Chinese American, Japanese American, and non-Mexican American Latino women. RESEARCH DESIGN AND METHODS: Homeostasis model assessments (HOMAs) of insulin sensitivity (HOMA%S) and beta-cell function (HOMA%beta) were used. Stepwise multivariable ethnic-specific ANCOVA models were used to compare HOMA%S and HOMA%beta between non-Hispanic whites and each of the four ethnic groups. RESULTS: HOMA%S was lower in African Americans, Chinese Americans, and Japanese Americans when compared with non-Hispanic white women after correcting for waist circumference, presence of impaired fasting glucose, and site. Significant differences persisted only between African Americans and non-Hispanic whites after inclusion of triglycerides in the model. Triglycerides indirectly corrected for the differences in HOMA%S in the other two groups. There were no differences in HOMA%S between the non-Mexican-American Latinos and the non Hispanic whites. Japanese Americans and Chinese Americans had lower HOMA%beta than non-Hispanic whites, whereas African Americans had higher HOMA%beta than non Hispanic whites after correcting for confounders. HOMA%beta was similar between non-Mexican-American Latinos and non-Hispanic whites. CONCLUSIONS: These data suggest that type 2 diabetes prevention strategies for African-American women should initially target decreased insulin sensitivity, whereas strategies for Japanese-American and Chinese-American women may initially need to target both decreased insulin sensitivity and beta-cell function. Previous studies of Mexican American populations may not apply to non-Mexican-American Latino women. PMID- 14747214 TI - Dietary antioxidant intake and risk of type 2 diabetes. AB - OBJECTIVE: The intake of antioxidants was studied for its ability to predict type 2 diabetes. RESEARCH DESIGN AND METHODS: A cohort of 2,285 men and 2,019 women 40 69 years of age and free of diabetes at baseline (1967-1972) was studied. Food consumption during the previous year was estimated using a dietary history interview. The intake of vitamin C, four tocopherols, four tocotrienols, and six carotenoids was calculated. During a 23-year follow-up, a total of 164 male and 219 female incident cases occurred. RESULTS: Vitamin E intake was significantly associated with a reduced risk of type 2 diabetes. The relative risk (RR) of type 2 diabetes between the extreme quartiles of the intake was 0.69 (95% CI 0.51 0.94, P for trend = 0.003). Intakes of alpha-tocopherol, gamma-tocopherol, delta tocopherol, and beta-tocotrienol were inversely related to a risk of type 2 diabetes. Among single carotenoids, beta-cryptoxanthin intake was significantly associated with a reduced risk of type 2 diabetes (RR 0.58, 95% CI 0.44-0.78, P < 0.001). No association was evident between intake of vitamin C and type 2 diabetes risk. CONCLUSIONS: This study supports the hypothesis that development of type 2 diabetes may be reduced by the intake of antioxidants in the diet. PMID- 14747215 TI - Screening for type 2 diabetes and impaired glucose metabolism: the Australian experience. AB - OBJECTIVE: To assess the Australian protocol for identifying undiagnosed type 2 diabetes and impaired glucose metabolism. RESEARCH DESIGN AND METHODS: The Australian screening protocol recommends a stepped approach to detecting undiagnosed type 2 diabetes based on assessment of risk status, measurement of fasting plasma glucose (FPG) in individuals at risk, and further testing according to FPG. The performance of and variations to this protocol were assessed in a population-based sample of 10,508 Australians. RESULTS: The protocol had a sensitivity of 79.9%, specificity of 79.9%, and a positive predictive value (PPV) of 13.7% for detecting undiagnosed type 2 diabetes and sensitivity of 51.9% and specificity of 86.7% for detecting impaired glucose tolerance (IGT) or impaired fasting glucose (IFG). To achieve these diagnostic rates, 20.7% of the Australian adult population would require an oral glucose tolerance test (OGTT). Increasing the FPG cut point to 6.1 mmol/l (110 mg/dl) or using HbA(1c) instead of FPG to determine the need for an OGTT in people with risk factors reduced sensitivity, increased specificity and PPV, and reduced the proportion requiring an OGTT. However, each of these protocol variations substantially reduced the detection of IGT or IFG. CONCLUSIONS: The Australian screening protocol identified one new case of diabetes for every 32 people screened, with 4 of 10 people screened requiring FPG measurement and 1 in 5 requiring an OGTT. In addition, 1 in 11 people screened had IGT or IFG. Including HbA(1c) measurement substantially reduced both the number requiring an OGTT and the detection of IGT or IFG. PMID- 14747216 TI - Trunk fat and leg fat have independent and opposite associations with fasting and postload glucose levels: the Hoorn study. AB - OBJECTIVE: Waist and hip circumferences have been shown to have independent and opposite associations with glucose levels. Waist circumference is positively associated with glucose levels, whereas hip circumference is negatively associated. It is unclear which tissues are involved in the pathophysiological mechanism causing these associations. The main goal was to determine which tissue in the trunk and legs, fat or lean tissue, is associated with measures of glucose metabolism. RESEARCH DESIGN AND METHODS: In 623 participants of the third examination of the Hoorn Study, whole-body dual-energy X-ray absorptiometry was performed to determine fat and lean soft-tissue mass in the trunk and legs. Fasting and 2-h postload glucose levels after 75-g oral glucose tolerance test (OGTT) were determined. After exclusion of known diabetic patients, cross sectional analyses were performed in 275 men aged 60-87 years (140 with normal glucose metabolism, 92 with impaired glucose metabolism; and 43 with diabetes) and in 281 women (148 with normal glucose metabolism, 90 with impaired glucose metabolism, and 43 with diabetes). RESULTS: Greater trunk fat mass was associated with higher glucose levels after adjustment for age, trunk lean mass, leg lean mass, and leg fat mass. Standardized beta (95% CI) in men were 0.44 (0.25-0.64) for fasting and 0.41 (0.22-0.60) for postload glucose. For women, these values were 0.49 (0.35-0.63) and 0.47 (0.33-0.61), respectively. In contrast, in the same regression models, a larger leg fat mass was associated with lower glucose levels. Standardized beta in men were -0.24 (-0.43 to -0.05) and -0.12 (-0.31 to 0.07) and in women -0.24 (-0.37 to -0.10) and -0.27 (-0.40 to -0.13) for fasting and postload glucose, respectively. In these models, larger leg lean mass was also associated with lower glucose levels but was only statistically significant in men. CONCLUSIONS: If trunk fat is taken into account, accumulation of fat in the legs seems to be protective against a disturbed glucose metabolism, particularly in women. Further research is needed to unravel underlying pathophysiological mechanisms. PMID- 14747217 TI - Obesity and the development of insulin resistance and impaired fasting glucose in black and white adolescent girls: a longitudinal study. AB - OBJECTIVE: Age at onset of type 2 diabetes has decreased during the past 20 years, especially in black women. Studies of factors associated with insulin resistance and hyperglycemia in preadolescent and adolescent populations are essential to understanding diabetes development. RESEARCH DESIGN AND METHODS: The National Heart, Lung, and Blood Institute (NHLBI) Growth and Health Study (NGHS) is a 10-year cohort study of the development of obesity in black and white girls. Two NGHS centers examined the associations of obesity, puberty, and race with fasting insulin, glucose, and homeostasis model assessment of insulin resistance (HOMA-IR; a calculated index of insulin resistance) measures at 9-10 years of age (baseline) and 10 years later. RESULTS: Black girls had greater baseline and year 10 BMI than white girls, with a greater 10-year incidence of obesity. BMI-insulin correlations were positive in both black and white girls at both visits, but insulin remained higher in black girls after controlling for BMI. In black girls, insulin and HOMA-IR were higher in the prepubertal period (before the emergence of racial differences in BMI), increased more during puberty, and decreased less with its completion. Baseline BMI predicted year-10 glucose and the development of impaired fasting glucose (IFG) in black girls. In white girls, the rate of BMI increase during follow-up predicted these outcomes. The 10-year incidence of diabetes in black girls was 1.4%. CONCLUSIONS: Black-white differences in insulin resistance are not just a consequence of obesity, but precede the pubertal divergence in BMI. The development of IFG appears to be a function of the rate of increase of BMI in white girls and early obesity in black girls. PMID- 14747218 TI - Problems paying out-of-pocket medication costs among older adults with diabetes. AB - OBJECTIVE: To identify problems faced by older adults with diabetes due to out-of pocket medication costs. RESEARCH DESIGN AND METHODS: In this cross-sectional national survey of 875 adults with diabetes treated with hypoglycemic medication, respondents reported whether they had underused prescription medications due to cost pressures or had experienced other financial problems associated with medication costs such as forgoing basic necessities. Respondents also described their interactions with clinicians about medication costs. RESULTS: A total of 19% of respondents reported cutting back on medication use in the prior year due to cost, 11% reported cutting back on their diabetes medications, and 7% reported cutting back on their diabetes medications at least once per month. Moreover, 28% reported forgoing food or other essentials to pay medication costs, 14% increased their credit card debt, and 10% borrowed money from family or friends to pay for their prescriptions. Medication cost problems were especially common among respondents who were younger, had higher monthly out-of-pocket costs, and had no prescription drug coverage. In general, few respondents, including those reporting medication cost problems, reported that their health care providers had given them information or other assistance to address medication cost pressures. CONCLUSIONS: Out-of-pocket medication costs pose a significant burden to many adults with diabetes and contribute to decreased treatment adherence. Clinicians should actively identify patients with diabetes who are facing medication cost pressures and assist them by modifying their medication regimens, helping them understand the importance of each prescribed medication, providing information on sources of low-cost drugs, and linking patients with coverage programs. PMID- 14747219 TI - Maternal anxiety associated with newborn genetic screening for type 1 diabetes. AB - OBJECTIVE: To describe maternal anxiety associated with newborn genetic screening for type 1 diabetes during the first year after risk notification. RESEARCH DESIGN AND METHODS: Mothers of at-risk infants (n = 435), identified through newborn genetic screening as part of the Prospective Assessment of Newborn for Diabetes Autoimmunity (PANDA) study, were administered a short form of the State Trait Anxiety Inventory (STAI) during telephone interviews approximately 3.5 weeks, 4 months, and 1 year after risk notification. Statistical analyses were conducted to examine predictors of maternal anxiety at each interview as well as changes in anxiety over time. RESULTS: For the total sample, initial state STAI scores were not elevated and declined further over time. However, Hispanic mothers, those with low levels of education, those who overestimated the child's risk for diabetes, and mothers of infants in the highest risk group exhibited significantly elevated initial state STAI scores. At 4 months, higher state STAI scores were associated with higher initial state STAI scores, single parent status, having an infant with a first-degree relative with diabetes, and overestimation of the child's actual risk. Initial and 4-month STAI scores remained predictive of STAI scores at 1 year. In addition, single mothers and mothers of female children reported higher STAI state scores 1 year after risk notification. CONCLUSIONS: For most mothers, newborn genetic screening to identify infants at increased risk for type 1 diabetes is not associated with significantly elevated maternal anxiety; anxiety further dissipates over time. However, anxiety levels vary considerably as a function of maternal ethnic status, education, marital status, maternal estimation of infant risk, and sex of the child and may be significantly elevated in some women. PMID- 14747220 TI - Quality of care and outcomes in type 2 diabetic patients: a comparison between general practice and diabetes clinics. AB - OBJECTIVE: The role of general practice and diabetes clinics in the management of diabetes is still a matter of debate. Methodological flaws in previous studies may have led to inaccurate conclusions when comparing the care provided in these different settings. We compared the care provided to type 2 diabetic patients attending diabetes outpatient clinics (DOCs) or being treated by a general practitioner (GP) using appropriate statistical methods to adjust for patient case mix and physician-level clustering. RESEARCH DESIGN AND METHODS: We prospectively evaluated the process and intermediate outcome measures over 2 years in a sample of 3,437 patients recruited by 212 physicians with different specialties practicing in 125 DOCs and 103 general practice offices. Process measures included frequency of HbA(1c), lipids, microalbuminuria, and serum creatinine measurements and frequency of foot and eye examinations. Outcome measures included HbA(1c), blood pressure, and total and LDL cholesterol levels. RESULTS: Differences for most process measures were statistically significantly in favor of DOCs. The differences were more marked for patients who were always treated by the same physician within a DOC and if that physician had a specialty in diabetology. Less consistent differences in process measures were detected when patients followed by GPs were compared with those followed by physicians with a specialty other than diabetology. As for the outcomes considered, patients attending DOCs attained better total cholesterol levels, whereas no major differences emerged in terms of metabolic control and blood pressure levels between DOCs and GPs. Physicians' specialties were not independently related to patient outcomes. CONCLUSIONS: Being followed always by the same physician in a DOC, particularly if the physician had a specialty in diabetes, ensured better quality of care in terms of process measures. In the short term, care provided by DOCs was also associated with better intermediate outcome measures, such as total cholesterol levels. PMID- 14747221 TI - Health-related quality of life and health-adjusted life expectancy of people with diabetes in Ontario, Canada, 1996-1997. AB - OBJECTIVE: To estimate the burden of illness from diabetes using a population health survey linked to a population-based diabetes registry. RESEARCH DESIGN AND METHODS: Measures of health-related quality of life (HRQOL) from the 1996/97 Ontario Health Survey (n = 35,517) were combined with diabetes prevalence and mortality data from the Ontario Diabetes Database (n = 487,576) to estimate the impact of diabetes on life expectancy, health-adjusted life expectancy (HALE), and HRQOL. RESULTS: Life expectancy of people with diabetes was 64.7 and 70.7 years for men and women, respectively-12.8 and 12.2 years less than that for men and women without diabetes. Diabetes had a large impact on instrumental and basic activities of daily living, more so than on functional health. HALE was 58.3 and 62.7 years, respectively, for men and women-11.9 and 10.7 years less than that of men and women without diabetes. Eliminating diabetes would increase Ontario life expectancy by 2.8 years for men and 2.6 years for women; HALE would increase by 2.7 and 3.2 years for men and women, respectively. CONCLUSIONS: The burden of illness from diabetes in Ontario is considerable. Efforts to reduce diabetes would likely result in a "compression of morbidity." An approach of estimating diabetes burden using linked data sources provides a robust approach for the surveillance of diabetes. PMID- 14747222 TI - Health-related quality of life and metabolic control in children with type 1 diabetes: a prospective cohort study. AB - OBJECTIVE: To assess change in health-related quality of life (HRQOL) in children with diabetes over 2 years and determine its relationship to change in metabolic control. RESEARCH DESIGN AND METHODS: In 1998, parents of children aged 5-18 years attending a tertiary diabetes clinic reported their child's HRQOL using the Child Health Questionnaire PF-50. Those aged 12-18 years also self-reported their HRQOL using the analogous Child Health Questionnaire CF-80. HbA(1c) levels were recorded. In 2000, identical measures were collected for those who were aged < or =18 years and still attending the clinic. RESULTS: Of 117 eligible subjects, 83 (71%) participated. Parents reported no significant difference in children's HRQOL at baseline and follow-up. However, adolescents reported significant improvements on the Family Activities (P < 0.001), Bodily Pain (P = 0.04), and General Health Perceptions (P = 0.001) scales and worsening on the Behavior (P = 0.04) scale. HbA(1c) at baseline and follow-up were strongly correlated (r = 0.57). HbA(1c) increased significantly (mean 7.8% in 1998 vs. 8.5% in 2000; P < 0.001), with lower baseline HbA(1c) strongly predicting an increase in HbA(1c) over the 2 years (r(2) = 0.25, P < 0.001). Lower parent-reported Physical Summary and adolescent-reported Physical Functioning scores at baseline also predicted increasing HbA(1c). Poorer parent-reported Psychosocial Summary scores were related to higher HbA(1c) at both times but did not predict change in HbA(1c). CONCLUSIONS: Changes in parent and adolescent reports of HRQOL differ. Better physical functioning may protect against deteriorating HbA(1c), at least in the medium term. While the HRQOL of children with diabetes does not appear to deteriorate over time, we should not be complacent, as it is consistently poorer than that of their healthy peers. PMID- 14747223 TI - Diabetes, major depression, and functional disability among U.S. adults. AB - OBJECTIVE: The goal of this study was to determine prevalence and odds of functional disability in individuals with diabetes and comorbid major depression compared with individuals with either diabetes or major depression alone. RESEARCH DESIGN AND METHODS: Data on 30,022 adults aged > or = 18 years from the 1999 National Health Interview Survey (NHIS) were analyzed. Four disease categories were created: no diabetes and no major depression, major depression alone, diabetes alone, and diabetes and comorbid major depression. Prevalence of functional disability was calculated for each disease category. Multiple logistic regression was used to determine the odds and correlates of functional disability by disease category controlling for age, sex, race/ethnicity, education, income, census region, and disability-associated comorbidity. STATA was used for all analyses to account for the complex survey design of NHIS. RESULTS: Irrevalence of functional disability by disease category was as follows: no diabetes and no major depression (24.5%); major depression (51.3%); diabetes (58.1%); and diabetes and comorbid major depression (77.8%). With no diabetes and no major depression as reference and after adjusting for relevant covariates, the odds of functional disability was 3.00 (95% CI 2.62-3.42) for major depression, 2.42 (2.10-2.79) for diabetes, and 7.15 (4.53-11.28) for diabetes and comorbid major depression. CONCLUSIONS: Individuals with diabetes and comorbid major depression have higher odds of functional disability compared with individuals with either diabetes or major depression alone. Additional studies are needed to establish a causal relationship. PMID- 14747224 TI - Depressive symptoms and the risk of type 2 diabetes: the Atherosclerosis Risk in Communities study. AB - OBJECTIVE: The goal of this study was to determine whether depressive symptoms predict type 2 diabetes. RESEARCH DESIGN AND METHODS: We analyzed data on depressive symptoms (including recent fatigue, sleep disturbance, feelings of hopelessness, loss of libido, and increased irritability) in a longitudinal, biracial cohort study of 11,615 initially nondiabetic adults aged 48-67 years, who were subsequently followed for 6 years for the development of type 2 diabetes. RESULTS: At baseline, depressive symptoms were positively associated with BMI, fasting insulin, systolic blood pressure, caloric intake, physical inactivity, and current smoking (all P < 0.05). In prospective analyses, after adjusting for age, race, sex, and education, individuals in the highest quartile of depressive symptoms had a 63% increased risk of developing diabetes compared with those in the lowest quartile (relative hazard [RH] 1.63, 95% CI 1.31-2.02). This relation persisted after adjustment for stress-associated lifestyle factors (smoking, physical activity, caloric intake, and adiposity) (1.28, 1.02-1.60) and metabolic covariates (fasting insulin and glucose, lipids, blood pressure, and adiposity) (1.38, 1.10-1.73). CONCLUSIONS: In this cohort, depressive symptoms predicted incident type 2 diabetes. This relation is only partially explained by demographic, metabolic, and lifestyle factors. Possible neuroendocrine mediators of the stress-obesity-diabetes relationship require further evaluation in prospective cohort studies that use an established tool to assess depression and incorporate neurohormonal measurements. PMID- 14747225 TI - Efficacy of Ipomoea batatas (Caiapo) on diabetes control in type 2 diabetic subjects treated with diet. AB - OBJECTIVE: To investigate the tolerability, efficacy, and mode of action of Caiapo, an extract of white sweet potatoes, on metabolic control in type 2 diabetic patients. RESEARCH DESIGN AND METHODS: A total of 61 type 2 diabetic patients treated by diet were given 4 g Caiapo (n = 30; mean age 55.2 +/- 2.1 years; BMI 28.0 +/- 0.4 kg/m(2)) or placebo (n = 31; mean age 55.6 +/- 1.5 years; BMI 27.6 +/- 0.3 kg/m(2)) once daily for 12 weeks. Each subject underwent a 75-g oral glucose tolerance test (OGTT) at baseline and after 1, 2, and 3 months to assess 2-h glucose levels. Additionally, fasting blood glucose, HbA(1c), total cholesterol, and triglyceride levels were measured. RESULTS: After treatment with Caiapo, HbA(1c) decreased significantly (P < 0.001) from 7.21 +/- 0.15 to 6.68 +/ 0.14%, whereas it remained unchanged (P = 0.23) in subjects given placebo (7.04 +/- 0.17 vs. 7.10 +/- 0.19%). Fasting blood glucose levels decreased (P < 0.001) in the Caiapo group (143.7 +/- 1.9 vs. 128.5 +/- 1.7 mg/dl) and did not change in the placebo group (144.3 +/- 1.9 vs. 138.2 +/- 2.1 mg/dl; P = 0.052). A decrease in body weight was observed in both the placebo group (P = 0.0027) and in the Caiapo group (P < 0.0001), probably due to a better- controlled lifestyle. In the Caiapo group, body weight was related to the improvement in glucose control (r = 0.618; P < 0.0002). Two-hour glucose levels were significantly (P < 0.001) decreased in the Caiapo group (193.3 +/- 10.4 vs. 162.8 +/- 8.2 mg/dl) compared with the placebo group (191.7 +/- 9.2 vs. 181.0 +/- 7.1 mg/dl). Mean cholesterol at the end of the treatment was significantly lower in the Caiapo group (214.6 +/ 11.2 mg/dl) than in the placebo group (248.7 +/- 11.2 mg/dl; P < 0.05). No significant changes in triglyceride levels or blood pressure were observed, and Caiapo was well tolerated without significant adverse effects. CONCLUSIONS: This study confirms the beneficial effects of Caiapo on plasma glucose as well as cholesterol levels in patients with type 2 diabetes. For the first time, the long term efficacy of Caiapo on glucose control was demonstrated by the observed decrease in HbA(1c). Thus, the neutraceutical Caiapo seems to be a useful agent in the treatment of type 2 diabetes. PMID- 14747226 TI - Comparison of the [13C]glucose breath test to the hyperinsulinemic-euglycemic clamp when determining insulin resistance. AB - OBJECTIVE: With increasing emphasis on the recognition of the metabolic syndrome and early type 2 diabetes, a clinically useful measure of insulin resistance is desirable. The purpose of this study was to evaluate whether an index of glucose metabolism, as measured by (13)CO(2) generation from ingested [(13)C]glucose, would correlate with indexes from the hyperinsulinemic-euglycemic clamp. RESEARCH DESIGN AND METHODS: A total of 26 subjects with varying degrees of insulin sensitivity underwent both the [(13)C]glucose breath test and the hyperinsulinemic-euglycemic clamp. Results from the [(13)C]glucose breath test were compared with measures of insulin sensitivity from the glucose clamp as well as with other commonly used indexes of insulin sensitivity. RESULTS: There was a strong correlation between the [(13)C]glucose breath test result and the glucose disposal rate (r = 0.69, P < 0.0001) and insulin sensitivity index (r = 0.69, P < 0.0001) from the insulin clamp. The magnitude of these correlations compared favorably with QUICKI and were superior to the homeostasis model assessment. CONCLUSIONS: The [(13)C]glucose breath test may provide a useful noninvasive assessment of insulin sensitivity. PMID- 14747227 TI - Cardiac abnormalities in diabetic patients with neuropathy: effects of aldose reductase inhibitor administration. AB - OBJECTIVE: The goal of this study was to determine whether treatment with an aldose reductase inhibitor (ARI) has beneficial effects on asymptomatic cardiac abnormalities in diabetic patients with neuropathy. RESEARCH DESIGN AND METHODS: Diabetic subjects with neuropathy (n = 81) with either a low diastolic peak filling rate or impaired augmentation of left ventricular (LV) ejection fraction (LVEF) during maximal bicycle exercise were identified by gated radionuclide ventriculography. Coronary artery disease, left ventricular hypertrophy, and valvular heart disease were excluded by clinical evaluation, myocardial perfusion imaging, and echocardiography. Subjects were randomized to receive blinded treatment with either the placebo or the ARI zopolrestat 500 or 1,000 mg daily for 1 year. RESULTS: After 1 year of ARI treatment, there were increases in resting LVEF (P < 0.02), cardiac output (P < 0.03), LV stroke volume (P < 0.004), and exercise LVEF (P < 0.001). In placebo-treated subjects, there were decreases in exercise cardiac output (P < 0.03), stroke volume (P < 0.02), and end diastolic volume (P < 0.04). Exercise LVEF increased with ARI treatment independent of blood pressure, insulin use, or the presence of baseline abnormal heart rate variability. There was no change in resting diastolic filling rates in either group. CONCLUSIONS: Diabetic patients with neuropathy have LV abnormalities that can be stabilized and partially reversed by ARI treatment. PMID- 14747228 TI - Intensification of therapeutic approaches reduces mortality in diabetic patients with acute myocardial infarction: the Munich registry. AB - OBJECTIVE: The myocardial infarction (MI) registry of the Academic Schwabing Hospital, Munich, investigates the hospital course of diabetic and nondiabetic patients with acute MI. The aim of this study was to improve quality care management and to compare hospital mortality and therapeutic approaches (i.e., PTCA, stenting, GPIIb/IIIa receptor antagonists, glucose-insulin infusion). RESEARCH DESIGN AND METHODS: Data of diabetic patients and nondiabetic patients were analyzed. All diabetic and nondiabetic subjects who were admitted in 1999 and 2001 were included: 1999, 126 (38%) diabetic and 204 (62%) nondiabetic patients; 2001, 91 (31%) diabetic and 205 (59%) nondiabetic patients. RESULTS: In 1999, coronary angiography (P < 0.01), percutaneous transluminal coronary angioplasty (PTCA) (P < 0.001), and stenting (P < 0.001) were performed less frequently in diabetic than in nondiabetic patients. During this period, total hospital mortality (29 vs. 16%, P < 0.01) and mortality within 24 h after admission (14 vs. 5%, P = 0.01) were higher in diabetic than in nondiabetic patients. In 2001, frequencies of coronary angiography, PTCA, and stenting were increased in diabetic patients (P < 0.001 vs. 1999), and the interventions were comparable with those performed in nondiabetic patients. Furthermore, glucose insulin infusion was administered in 46% of diabetic subjects. In 2001, total hospital mortality decreased to 17% in diabetic subjects (P = 0.028 vs. 1999) and mortality within 24 h after admission declined to 4% (P = 0.027 vs. 1999). Logistic regression analysis revealed that an increase in the number of therapeutic approaches (also when adjusted for clinical variables) is associated with a reduction in mortality of diabetic patients with acute MI (adjusted odds ratio 0.14, P < 0.0001). CONCLUSIONS: Intensification of multiple advanced therapeutic strategies in diabetic patients with acute MI enables a substantial reduction in hospital mortality. The enforcement leads to rates of hospital mortality that are comparable to those of nondiabetic patients. PMID- 14747229 TI - Implementation of a safe and effective insulin infusion protocol in a medical intensive care unit. AB - OBJECTIVE: In a recent randomized controlled trial, lowering blood glucose levels to 80-110 mg/dl improved clinical outcomes in critically ill patients. In that study, the insulin infusion protocol (IIP) used to normalize blood glucose levels provided valuable guidelines for adjusting insulin therapy. In our hands, however, ongoing expert supervision was required to effectively manage the insulin infusions. This work describes our early experience with a safe, effective, nurse-implemented IIP that provides detailed insulin dosing instructions and requires minimal physician input. RESEARCH DESIGN AND METHODS: We collected data from 52 medical intensive care unit (MICU) patients who were placed on the IIP. Blood glucose levels were the primary outcome measurement. Relevant clinical variables and insulin requirements were also recorded. MICU nurses were surveyed regarding their experience with the IIP. RESULTS: To date, our IIP has been employed 69 times in 52 patients admitted to an MICU. Using the IIP, the median time to reach target blood glucose levels (100-139 mg/dl) was 9 h. Once blood glucose levels fell below 140 mg/dl, 52% of 5,808 subsequent hourly blood glucose values fell within our narrow target range; 66% within a "clinically desirable" range of 80-139 mg/dl; and 93% within a "clinically acceptable" range of 80-199 mg/dl. Only 20 (0.3%) blood glucose values were <60 mg/dl, none of which resulted in clinically significant adverse events. In general, the IIP was readily accepted by our MICU nursing staff, most of whom rated the protocol as both clinically effective and easy to use. CONCLUSIONS: Our nurse-implemented IIP is safe and effective in improving glycemic control in critically ill patients. PMID- 14747230 TI - Anti-inflammatory and anticoagulant effects of pravastatin in patients with type 2 diabetes. AB - OBJECTIVE: Type 2 diabetes is associated with increased plasma concentrations of coagulation and inflammation markers. Different studies have shown that treatment with hydroxymethylglutaryl-CoA reductase inhibitors (statins) is associated with antithrombotic and anti-inflammatory effects in addition to a cholesterol lowering effect. Our objective was to evaluate the effect of pravastatin (40 mg/day) on coagulation and inflammation markers in type 2 diabetic patients. RESEARCH DESIGN AND METHODS: This was an open, randomized, crossover study designed with an 8-week intervention period. The study group was comprised of 50 patients with type 2 diabetes (median HbA(1c) 7.1%) and serum total cholesterol of 5-10 mmol/l. We evaluated plasma levels of fibrinogen, F1 + 2, D-dimer, soluble tissue factor (sTF), von Willebrand Factor antigen (vWFag), and C reactive protein (CRP) in blood samples drawn after fasting on day 1 and after 8 and 16 weeks. RESULTS: Significant reductions of total cholesterol (-22%; P < 0.001), LDL cholesterol (-32%; P < 0.001), and triglycerides (-10%; P < 0.05) were achieved after 8 weeks of treatment with pravastatin. In addition, significant reductions of plasma levels of F1 + 2 (-4.4%; P < 0.05), vWFag ( 5.3%; P < 0.05), and sTF (-3.4%; P < 0.05) were observed after treatment with pravastatin. Furthermore, plasma levels of CRP were also significantly reduced ( 13%; P < 0.05). Levels of fibrinogen and D-dimer did not decrease after treatment with pravastatin. CONCLUSIONS: The results indicated that pravastatin reduces levels of coagulation and inflammation markers in type 2 diabetic patients. These antithrombotic and anti-inflammatory effects of treatment with statins could play a role in reducing cardiovascular complications in type 2 diabetic patients. PMID- 14747231 TI - Effectiveness of different types of footwear insoles for the diabetic neuropathic foot: a follow-up study. AB - OBJECTIVE: To compare the effectiveness of different types of footwear insoles in the diabetic neuropathic foot. RESEARCH DESIGN AND METHODS: A sample of 241 consecutive diabetic patients (158 men and 83 women, age 57.5 +/- 9.6 years [mean +/- SD], and mean duration of diabetes 12.3 +/- 7.2 years) attending the foot clinic with previous foot ulceration and those considered at high risk of foot ulceration were included in the study. The study groups consisted of group 1, patients provided with sandals with insoles made with microcellular rubber (n = 100); group 2, with sandals with polyurethane foam (n = 59); group 3, with molded insoles (n = 32); and group 4, with their own footwear containing leather board insoles (n = 50). Neuropathy status was assessed using a biothesiometer. Plantar pressure was measured using the RS Scan inshoe pressure measurement system. Data obtained from the metatarsal heads were used as the peak pressure. The state of the sandals was assessed after 9 months. The patients were considered to have had an ulcer relapse when either a new ulcer appeared at the site of a previous one or a new foot ulcer appeared in a different area. RESULTS: Patients who were using therapeutic footwear showed lower foot pressure (group 1, 6.9 +/- 3.6; group 2, 6.2 +/- 3.9; and group 3, 6.8 +/- 6.1 kPa; P = 0.0001), while those who used the nontherapeutic footwear showed an increased foot pressure (group 4, 40.7 +/- 20.5 kPa; P = 0.008). The occurrence of new lesions was significantly higher in patients in group 4 (33%) when compared with that of all other groups (4%). CONCLUSION: Therapeutic footwear is useful to reduce new ulceration and consequently the amputation rate in the diabetic population. PMID- 14747232 TI - Establishment of blood glucose monitoring system using the internet. AB - OBJECTIVE: The Internet is used worldwide as a communication tool. To improve the quality of diabetes control, we investigated the effectiveness of an Internet based blood glucose monitoring system (IBGMS) on controlling the changes in HbA(1c) levels. RESEARCH DESIGN AND METHODS: We conducted a randomized clinical trial involving 110 patients who visited the outpatient clinic at the Kangnam St. Mary's Hospital for 3 months. The study subjects were treated with IBGMS for 12 weeks, and the control group received the usual outpatient management over the same period. HbA(1c) and other laboratory tests were performed twice, once at the beginning of the study and again at the end of the study. RESULTS: The test results from the beginning of the study established that there were no significant differences between the two groups with respect to age, sex, diabetes duration, BMI, blood pressure, HbA(1c), and other laboratory data. On follow-up examination 12 weeks later, HbA(1c) levels were significantly decreased from 7.59 to 6.94% within the intervention group (P < 0.001). At the end of the study, HbA(1c) levels in the intervention group were significantly lower than in the control group after adjusting the baseline HbA(1c) (6.94 vs. 7.62%; P < 0.001, respectively). Among patients with baseline HbA(1c) <7.0%, the patients in the intervention group had lower HbA(1c) than those in the control group (6.38 vs. 6.99%; P < 0.05). Among the patients with a baseline HbA(1c) > or = 7.0%, the difference between the two groups appeared more obvious: HbA(1c) levels at the end of the study were 8.12%. CONCLUSIONS: This new IBGMS resulted in a significant reduction of HbA(1c) during the study period. We propose that this IBGMS be used as a method for improving diabetes control. PMID- 14747233 TI - In type 2 diabetes, rosiglitazone therapy for insulin resistance ameliorates endothelial dysfunction independent of glucose control. AB - OBJECTIVE: Insulin resistance is an independent risk factor for arteriosclerosis and cardiovascular mortality. However, the mechanism by which insulin resistance contributes to arteriosclerosis is unknown. Conceivably, endothelial dysfunction could be involved. Therefore, we asked whether therapy for insulin resistance ameliorates any endothelial dysfunction. RESEARCH DESIGN AND METHODS: We performed a double-blind cross-over trial of 12 patients with recently diagnosed type 2 diabetes. They received rosiglitazone 4 mg b.i.d. for 12 weeks and nateglinide 60 mg b.i.d. for the same number of weeks in random order. To assess the degree of endothelial dysfunction, we used venous occlusion plethysmography. We studied vasodilation in response to acetylcholine (ACh) with and without exogenous insulin. The agents were infused into the brachial artery. Furthermore, we determined insulin resistance by euglycemic clamp. RESULTS: Glycemic control was comparable under rosiglitazone and nateglinide. Rosiglitazone ameliorated insulin resistance by 60% compared with nateglinide. ACh response was significantly increased after rosiglitazone treatment (maximum forearm blood flow 12.8 +/- 1.3 vs. 8.8 +/- 1.3 ml/100 ml after rosiglitazone and nateglinide, respectively; P < 0.05) but did not attain the level of healthy control subjects (14.0 +/- 0.7 ml/100 ml). Coinfusion of exogenous insulin increased ACh response further in the rosiglitazone group. N-monomethyl-L-arginine-acetate (L-NMMA), an antagonist of nitric oxide synthase, largely prevented the increased vasodilation after rosiglitazone, regardless of the presence or absence of insulin. Insulin sensitivity and blood flow response were found to be correlated (P < 0.01). CONCLUSIONS: Insulin resistance is a major contributor toward endothelial dysfunction in type 2 diabetes. Both endothelial dysfunction and insulin resistance are amenable to treatment by rosiglitazone. PMID- 14747234 TI - Elevated white blood cell count in subjects with impaired glucose tolerance. AB - OBJECTIVE: Impaired glucose tolerance (IGT) and impaired fasting glucose (IFG) differ in their risk of all-cause and cardiovascular mortality, but previous cross-sectional studies have suggested little difference in their levels of lipids or blood pressure. We compared the white blood cell (WBC) count between subjects with IFG and IGT. RESEARCH DESIGN AND METHODS: The subjects were 4,720 nondiabetic Japanese men aged 24-84 years. Based on the 75-g oral glucose tolerance test, the subjects were classified into the following four groups: normal fasting glucose/normal glucose tolerance (n = 3,753), isolated IFG (n = 290), isolated IGT (n = 476), and IFG/IGT (n = 201). We compared the WBC count among the four groups and investigated variables that showed a significant association with the WBC count. RESULTS: The isolated IGT group had a significantly higher WBC count than the isolated IFG group (6,530 vs. 6,210/mm(3), P < 0.05). By stepwise analyses, age, triglycerides, HDL cholesterol, fasting insulin, and 2-h postchallenge plasma glucose (PG) showed an independent association with the WBC count (adjusted R(2) = 0.057). In the analysis stratified by smoking status, the WBC count was independently associated with 2-h PG and triglycerides, irrespective of smoking status. CONCLUSIONS: Individuals with isolated IGT had a significantly higher WBC count than those with isolated IFG. The WBC count was associated with 2-h PG and various components of the metabolic syndrome. PMID- 14747235 TI - Is presence of islet autoantibodies at birth associated with development of persistent islet autoimmunity? The Diabetes Autoimmunity Study in the Young (DAISY). AB - OBJECTIVE: To determine whether the presence of islet autoantibodies in the umbilical cord blood is predictive of subsequent development of islet autoimmunity. RESEARCH DESIGN AND METHODS: Cord blood sera from 1,118 subjects from the Diabetes Autoimmunity Study in the Young (DAISY) cohort, as well as their venous blood samples taken at follow-up clinic visits, were tested for GAD65 autoantibodies (GAAs), insulin autoantibodies (IAAs), and IA-2 autoantibodies (IA-2As). Venous blood samples taken from mothers of cord blood autoantibody-positive children were analyzed for the same autoantibodies. RESULTS: At least one of three islet autoantibodies was present in 42 (3.7%) of the cord blood samples tested. The presence of cord blood autoantibodies did not predict the subsequent development of islet autoimmunity (adjusted hazard ratio = 0.73 [0.09, 5.88]). Discordance between cord blood and corresponding maternal autoantibodies was seen in 3 of 36 infants. A strong correlation between levels of autoantibody in cord blood and maternal circulation was found for GAA (r(2) = 0.93, P < 0.001) and IAA (r(2) = 0.89, P < 0.001) but not IA-2A (r(2) = 0.05, P = 0.19). Cord blood autoantibodies in all but one subject disappeared by 9 months of age. CONCLUSIONS: The presence of cord blood autoantibodies is not predictive of subsequent development of islet autoimmunity. The majority of cord blood autoantibodies appear to result from maternal transmission. PMID- 14747236 TI - Leucine 7 to proline 7 polymorphism in the preproneuropeptide Y is associated with proteinuria, coronary heart disease, and glycemic control in type 1 diabetic patients. AB - OBJECTIVE: Neuropeptide Y is a potent vasoconstrictor thought to enhance the development of atherosclerosis. The leucine 7 to proline 7 (Leu7Pro) polymorphism, located in the signal peptide part of the human preproneuropeptide Y, has been associated with serum lipid levels, intima-media thickness of the common carotid arteries, and diabetic retinopathy in type 2 diabetic patients. Therefore, we investigated the impact of the Leu7Pro polymorphism on diabetic nephropathy, cardiovascular risk factors, and cardiovascular disease in type 1 diabetic patients. RESEARCH DESIGN AND METHODS: A total of 996 patients from the Finnish Diabetic Nephropathy study were studied in a case-control, cross sectional study. The carrier frequency of the Pro7 substitution was 13% in the entire study population. RESULTS: The Pro7 substitution was more common in patients with proteinuria than in those with a normal albumin excretion rate (16 vs. 11%, P < 0.05). Patients with the Pro7 allele had worse glycemic control (HbA(1c) 8.8 vs. 8.5%, P < 0.005), more coronary heart disease (CHD) (14 vs. 8%, P < 0.05), and higher serum triglycerides (1.65 vs. 1.35 mmol/l, P < 0.005) than patients with the wild-type genotype. There were no differences in the plasma neuropeptide Y levels between the patients with Pro7 compared with those with the wild-type genotype. The Leu7Pro polymorphism was independently associated with HbA(1c) (P < 0.001), proteinuria (P < 0.01), and CHD (P < 0.01) in multiple regression analyses. CONCLUSIONS: We conclude that the Leu7Pro polymorphism may contribute to the genetic susceptibility to diabetic nephropathy and CHD in type 1 diabetic patients, possibly by influencing glycemic control and triglycerides. PMID- 14747237 TI - Nondipping and its relation to glomerulopathy and hyperfiltration in adolescents with type 1 diabetes. AB - OBJECTIVE: To determine whether there is a relation between dipping/nondipping status and end-organ damage (measured as renal glomerulopathy) and long-term renal function in order to predict the development of nephropathy in normoalbuminuric patients with type 1 diabetes. RESEARCH DESIGN AND METHODS: Analysis of renal biopsy and ambulatory blood pressure measurements was done in relation to renal function tests performed during a 10-year period. Forty unselected patients (16 girls), with a mean age of 17.7 years and a mean duration of 10.7 years, were studied. The renal biopsies were examined by electron microscopy. Ambulatory blood pressure was monitored (Space Labs 90 207). Systolic nondippers were defined as a <7%, diastolic nondippers as a <14%, and mean arterial blood pressure (MAP) nondippers as a <12% fall in blood pressure during the night. Renal function was evaluated every other year by clearances of inulin (glomerular filtration rate [GFR]) and para-amino hippurate (effective renal plasma flow [ERPF]), and filtration fraction (GFR/ERPF) was calculated. Overnight urinary albumin excretion rate and long-term mean HbA(1c) were measured. RESULTS: MAP (27% of the patients) and diastolic nondippers (12%) had a significantly thicker basement membrane; larger mesangial matrix volume fraction; and higher long-term GFR, nighttime heart rate, and mean HbA(1c) than dippers. CONCLUSIONS: Nondipping status was related to more renal morphological changes and long-term hyperfiltration in normoalbuminuric adolescents and young adults, despite a short duration of type 1 diabetes. Nondipping status may be an early predictor of later nephropathy. PMID- 14747238 TI - Lipoprotein(a) is an independent risk factor for peripheral arterial disease in Chinese type 2 diabetic patients in Taiwan. AB - OBJECTIVE: To examine the association between lipoprotein(a) [Lp(a)] and peripheral arterial disease (PAD) and determine the optimal cutoff in Chinese type 2 diabetic patients in Taiwan. RESEARCH DESIGN AND METHODS: Serum Lp(a) was determined in 557 type 2 diabetic patients (243 men and 314 women) recruited consecutively from a diabetes clinic at the National Taiwan University Hospital. Ankle-brachial index (ABI) <0.9 was diagnosed as PAD (n = 45) and <0.8 as severe PAD (n = 20). Potential confounders included age, sex, BMI, waist-to-hip ratio (WHR), diabetes duration, insulin usage, smoking, hypertension, systolic and diastolic blood pressure, fasting plasma glucose (FPG), total cholesterol, triglycerides, and HDL and LDL cholesterol. RESULTS: The distribution of Lp(a) was right skewed and no significant differences for sex, WHR, insulin usage, smoking, hypertension, and systolic and diastolic blood pressure were observed. In men, log[Lp(a)] was correlated positively with age, duration, and total and LDL cholesterol (borderline significant, P < 0.1) and negatively with BMI, triglycerides, and FPG (P < 0.1). In women, log[Lp(a)] was correlated positively with total and LDL cholesterol and negatively with triglycerides and BMI (P < 0.1). ABI was significantly correlated with log[Lp(a)], especially in men or in patients with PAD. The optimal cutoff determined by discriminant analysis was 13.3 mg/dl. Patients with Lp(a) above this value had a 2.7-fold higher risk of PAD after multivariate adjustment. Lp(a) also significantly increased from no PAD to mild and severe PAD (17.1 +/- 14.4, 23.7 +/- 20.3, and 36.9 +/- 22.8 mg/dl, respectively, P < 0.001). CONCLUSIONS: Lp(a) is an independent risk factor for PAD in type 2 diabetic patients in Taiwan. The optimal cutoff is 13.3 mg/dl. PMID- 14747239 TI - Left ventricular mass increases with deteriorating glucose tolerance, especially in women: independence of increased arterial stiffness or decreased flow-mediated dilation: the Hoorn study. AB - OBJECTIVE: Type 2 diabetes and impaired glucose metabolism (IGM) are associated with an increased cardiovascular disease (CVD) risk. Increased left ventricular mass (LVM) is thought to increase CVD risk through several unfavorable cardiac changes. Type 2 diabetes and IGM are associated with increased LVM, but the underlying mechanism is unclear. We investigated the association between glucose tolerance status (GTS) and LVM and explored whether any such association could be mediated through increased arterial stiffness, impaired endothelial function, or the presence of atherosclerosis. RESEARCH DESIGN AND METHODS: We used ultrasound to measure LVM, carotid and femoral stiffness, carotid-femoral transit time, and flow-mediated vasodilation (FMD) and tonometry to estimate compliance and augmentation index. The study population (n = 780) consisted of 287 individuals with normal glucose metabolism (NGM), 179 with IGM, and 314 with type 2 diabetes, and the mean age was 68.4 years. RESULTS: In women, after adjusting for age, height, BMI, and mean arterial pressure, LVM increased significantly with deteriorating GTS (LVM 157 g in NGM, 155 g in IGM, and 169 g in type 2 diabetes; P for trend <0.018). Additional adjustment for arterial stiffness, FMD, or the presence of atherosclerosis did not materially alter the results, even though these variables were significantly associated with both GTS and LVM. Indexes of hyperglycemia/-insulinemia or insulin resistance explained at most 7% of the association between GTS and LVM. In men, no statistically significant associations were observed. CONCLUSIONS: Our data expand the conceptual view of the pathogenesis of GTS-related changes in LVM because we show that the increase in LVM in women is independent of increased arterial stiffness, impaired FMD, or the presence of atherosclerosis. In addition, we show that this increase in LVM is only minimally explained by indexes of hyperglycemia/-insulinemia or insulin resistance. Our data may, in part, explain the increased CVD risk seen in women with deteriorating GTS. PMID- 14747240 TI - Risk factors for coronary heart disease in type 1 diabetic patients in Europe: the EURODIAB Prospective Complications Study. AB - OBJECTIVE: The goal of the study was to examine risk factors in the prediction of coronary heart disease (CHD) and differences in men and women in the EURODIAB Prospective Complications Study. RESEARCH DESIGN AND METHODS: Baseline risk factors and CHD at follow-up were assessed in 2,329 type 1 diabetic patients without prior CHD. CHD was defined as physician-diagnosed myocardial infarction, angina pectoris, coronary artery bypass graft surgery, and/or Minnesota-coded ischemic electrocardiograms or fatal CHD. RESULTS: There were 151 patients who developed CHD, and the 7-year incidence rate was 8.0 (per 1,000 person-years) in men and 10.2 in women. After adjustment for age and/or duration of diabetes, the following risk factors were related to CHD in men: age, GHb, waist-to-hip ratio (WHR), HDL cholesterol, smoking, albumin excretion rate (AER), and autonomic neuropathy. The following risk factors were related to CHD in women: age, systolic blood pressure (BP), fasting triglycerides, AER, and retinopathy. Multivariate standardized Cox proportional hazards models showed that age (hazard ratio 1.5), AER (1.3 in men and 1.6 in women), WHR (1.3 in men), smoking (1.5 in men), fasting triglycerides (1.3 in women) or HDL cholesterol (0.74 in women), and systolic BP (1.3 in women) were predictors of CHD. CONCLUSIONS: This study supports the evidence for a strong predictive role of baseline albuminuria in the pathogenesis of CHD in type 1 diabetes. Furthermore, sex-specific risk factors such as systolic BP, fasting triglycerides (or HDL cholesterol), and WHR were found to be important in the development of CHD. PMID- 14747241 TI - Carbohydrate nutrition, insulin resistance, and the prevalence of the metabolic syndrome in the Framingham Offspring Cohort. AB - OBJECTIVE: The aim of this study was to examine the relation between carbohydrate related dietary factors, insulin resistance, and the prevalence of the metabolic syndrome in the Framingham Offspring Cohort. RESEARCH DESIGN AND METHODS: We examined cross-sectional associations between carbohydrate-related dietary factors, insulin resistance, and the prevalence of the metabolic syndrome in 2,834 subjects at the fifth examination (1991-1995) of the Framingham Offspring Study. Homeostasis model assessment of insulin resistance (HOMA-IR) was calculated using the following formula (fasting plasma insulin x plasma glucose)/22.5. The metabolic syndrome was defined using the National Cholesterol Education Program criteria. RESULTS: After adjustment for potential confounding variables, intakes of total dietary fiber, cereal fiber, fruit fiber, and whole grains were inversely associated, whereas glycemic index and glycemic load were positively associated with HOMA-IR. The prevalence of the metabolic syndrome was significantly lower among those in the highest quintile of cereal fiber (odds ratio [OR] 0.62; 95% CI 0.45-0.86) and whole-grain (0.67; 0.48-0.91) intakes relative to those in the lowest quintile category after adjustment for confounding lifestyle and dietary factors. Conversely, the prevalence of the metabolic syndrome was significantly higher among individuals in the highest relative to the lowest quintile category of glycemic index (1.41; 1.04-1.91). Total carbohydrate, dietary fiber, fruit fiber, vegetable fiber, legume fiber, glycemic load, and refined grain intakes were not associated with prevalence of the metabolic syndrome. CONCLUSIONS: Whole-grain intake, largely attributed to the cereal fiber, is inversely associated with HOMA-IR and a lower prevalence of the metabolic syndrome. Dietary glycemic index is positively associated with HOMA IR and prevalence of the metabolic syndrome. Given that both a high cereal fiber content and lower glycemic index are attributes of whole-grain foods, recommendation to increase whole-grain intake may reduce the risk of developing the metabolic syndrome. PMID- 14747242 TI - Adiponectin in youth: relationship to visceral adiposity, insulin sensitivity, and beta-cell function. AB - OBJECTIVE: Adiponectin is an adipose tissue protein that enhances insulin sensitivity and has antiatherogenic properties. The present study investigated the relationship of adiponectin levels in adolescents to 1) obesity and body fat distribution and 2) insulin sensitivity and the components of syndrome X. RESEARCH DESIGN AND METHODS: Twenty-three normal-weight and 26 obese adolescents had fasting adiponectin, lipid profile, and proinsulin measurements performed. Hepatic and peripheral insulin sensitivity were assessed with constant-rate [6,6 (2)H(2)]glucose infusion and a 3-h hyperinsulinemic-euglycemic clamp, respectively. Body composition was evaluated by dual-energy X-ray absorptiometry, and visceral adipose tissue (VAT) and subcutaneous adipose tissue were measured by computed tomography scan at the L(4)-L(5) level. RESULTS: Obese adolescents had approximately 50% lower adiponectin than normal-weight adolescents. Moreover, obese adolescents with high (111.8 +/- 9.3 cm(2)) versus low (55.4 +/- 2.1 cm(2)) VAT had lower adiponectin levels (6.2 +/- 0.9 vs. 9.0 +/- 1.0 microg/ml, P = 0.05). Plasma adiponectin correlated positively with peripheral and hepatic insulin sensitivity (r = 0.67, P < 0.001 and r = 0.54, P < 0.001, respectively) and HDL (r = 0.52, P < 0.001) and negatively with fasting proinsulin and the proinsulin-to-insulin ratio (r = -0.64, P < 0.001 and r = -0.43, P = 0.003, respectively). In a multiple regression analysis, adiponectin, independently and together with BMI, explained 73% (R(2) = 0.73, P < 0.001) of the variance in insulin sensitivity. Adiponectin, but not adiposity, was the significant independent determinant of the proinsulin-to-insulin ratio (R(2) = 0.18, P = 0.008) and of HDL (R(2) = 0.45, P < 0.001). CONCLUSIONS: In summary, hypoadiponectinemia in youth is a strong and independent correlate of insulin resistance, beta-cell dysfunction, visceral adiposity, and syndrome X. The antidiabetogenic and antiatherogenic properties of adiponectin are evident early in life and compromised in youth-onset obesity. PMID- 14747243 TI - Management of diabetes and hyperglycemia in hospitals. PMID- 14747244 TI - Lowering the cut point for impaired fasting glucose: where is the evidence? Where is the logic? PMID- 14747245 TI - Consensus development conference on antipsychotic drugs and obesity and diabetes. PMID- 14747246 TI - The 1st World Congress on the Insulin Resistance Syndrome. PMID- 14747247 TI - Never say never in medicine: confessions of an old dog. PMID- 14747248 TI - Dietary approaches to prevent the metabolic syndrome: quality versus quantity of carbohydrates. PMID- 14747249 TI - Relationship between periodontal disease and diabetic retinopathy. PMID- 14747250 TI - Alternative site testing at the earlobe tip: reliability of glucose measurements and pain perception. PMID- 14747251 TI - Effect of diabetes intervention programs on physical activity among migrant Mexican women with type 2 diabetes. PMID- 14747252 TI - Needle reuse and tip damage. PMID- 14747253 TI - Depression, diabetes, and glycemic control in Pima Indians. PMID- 14747254 TI - The Diabetes Treatment Satisfaction Questionnaire: a cross-cultural South African perspective. PMID- 14747255 TI - Improper insulin compliance may lead to hepatomegaly and elevated hepatic enzymes in type 1 diabetic patients. PMID- 14747256 TI - Acute hyperglycemia: implications for contrast-induced nephropathy during cardiac catheterization. PMID- 14747257 TI - Influence of the polymorphisms Tpr64Arg in the beta 3-adrenergic receptor gene and Pro12Ala in the PPAR gamma 2 gene on metabolic syndrome-related phenotypes in an indigenous population of the Brazilian Amazon. PMID- 14747258 TI - Elevated serum ferritin concentrations in a glucose-impaired population and in normal glucose tolerant first-degree relatives in familial type 2 diabetic pedigrees. PMID- 14747259 TI - Simultaneous bilateral facial palsy in a diabetic patient. PMID- 14747260 TI - Reduced fear of hypoglycemia in successful islet transplantation. PMID- 14747261 TI - A case of lipoatrophy with Lispro insulin without insulin pump therapy. PMID- 14747262 TI - Diabetes in a nonpancreatectomized child with nesidioblastosis. PMID- 14747263 TI - Oral glucose tolerance test evaluation with forearm and fingertip glucose measurements in pregnant women. PMID- 14747264 TI - Silent hypoglycemia presenting as dysesthesias. PMID- 14747265 TI - Exercise increases adiponectin levels and insulin sensitivity in humans. PMID- 14747266 TI - High glucose levels induce an increase in membrane antioxidants, in terms of vitamin E and coenzyme Q10, in children and adolescents with type 1 diabetes. PMID- 14747267 TI - Increased oxidative stress is associated with serum levels of triglyceride, insulin resistance, and hyperinsulinemia in Japanese metabolically obese, normal weight men. PMID- 14747268 TI - Efficacy of conversion from bedtime NPH insulin injection to once- or twice-daily injections of insulin glargine in type 1 diabetic patients using basal/bolus therapy. PMID- 14747269 TI - Memory impairments associated with postprandial hyperglycemia and glycemic control: comment on Greenwood et al. PMID- 14747271 TI - Off-loading in trials in neuropathic diabetic foot ulceration: no, it's not time for a paradigm shift. PMID- 14747274 TI - C-reactive protein and glycemic control in adults with diabetes: response to King et al. PMID- 14747275 TI - Autoimmune diabetes and the circle of tolerance. AB - The concept of immunological tolerance is central to our understanding of type 1 diabetes and the development of strategies for its prediction, prevention, and cure. Tolerance simply refers to the absence of an immune response. Most of us are born with an immune system that develops tolerance to all the other systems of our bodies as well as to the things that we eat. It is the loss of immunological tolerance that leads to autoimmunity. And when that autoimmune response directly or indirectly targets the beta-cell, type 1 diabetes is the result. In the U.S., 1 in 600 of us loses tolerance to pancreatic beta-cells. Interference with T-cell function after the loss of tolerance, as can be achieved with immunosuppressive drugs like cyclosporin, arrests the disease, but the cost in side effects is high. Clearly, stopping the loss of tolerance would be preferable. If we can stop the loss of tolerance, we can prevent the disease. We and many others have investigated both approaches. But what of the people who already have diabetes? For them a separate but related strategy, tolerance induction, is required. Specifically, islet transplantation tolerance induction holds out the promise of being able to cure the disease. This has been the ultimate goal of our laboratory's work for the past two decades. PMID- 14747276 TI - Chronic sympathetic activation: consequence and cause of age-associated obesity? AB - Primary aging in adult humans is associated with a progressive, tonic activation of the peripheral sympathetic nervous system (SNS). The purpose of this SNS activation and its physiological impact are, however, unknown. We hypothesize that the chronic stimulation of the SNS with aging is driven in part by a progressive accumulation of body fat. This "error" is sensed by the central nervous system via increases in adiposity-sensitive humoral signals (e.g., leptin, insulin) that cross the blood-brain barrier, activate subcortical areas involved in the regulation of energy balance (e.g., ventromedial hypothalamus), and stimulate SNS outflow to peripheral tissues. The SNS activation is intended to increase beta-adrenergic thermogenesis in order to expend excess energy as heat rather than by storage of fat. Recent evidence, however, indicates that these adjustments are not effective in augmenting energy expenditure with aging. Indeed, older sedentary adults demonstrate reduced, not increased, beta adrenergic stimulation of metabolic rate because of reduced tissue responsiveness, presumably mediated by SNS-induced impairment of beta-adrenergic signaling. As a result, age-associated SNS activation, initiated as a consequence of accumulating adiposity with the intent of preventing further fat storage, ironically, may in time evolve into a potential mechanism contributing to the development of obesity with aging. PMID- 14747277 TI - Suppression of endogenous glucose production by mild hyperinsulinemia during exercise is determined predominantly by portal venous insulin. AB - Hyperinsulinemia during exercise in people with diabetes requiring exogenous insulin is a major clinical problem. The aim of this study was to assess the significance of portal vein versus arterial insulin to hepatic effects of hyperinsulinemia during exercise. Dogs had sampling (artery, portal vein, and hepatic vein) and infusion (vena cava and portal vein) catheters and flow probes (hepatic artery and portal vein) implanted >16 days before a study. Protocols consisted of equilibration (-130 to -30 min), basal (-30 to 0 min), and treadmill exercise (0-150 min) periods. Somatostatin was infused and glucagon and insulin were replaced in the portal vein to achieve basal arterial and portal vein levels at rest and simulated levels during the first 60 min of exercise. From 60 to 150 min of exercise, the simulated insulin infusion was sustained (C; n = 7), modified to selectively create a physiologic increment in arterial insulin (Pe; n = 7), or altered to increase arterial insulin as in Pe but with a concomitant increase in portal insulin (PePo; n = 7). Euglycemic clamps were performed in all studies. Portal and arterial insulin were 15 +/- 2 and 4 +/- 1 micro U/ml (mean +/- SE of all groups), respectively, at t = 60 min in all groups. Insulin levels were unchanged for the remainder of the exercise period in C. Arterial insulin was increased from 3 +/- 1 to 14 +/- 2 micro U/ml, whereas portal insulin did not change in Pe after t = 60 min. Arterial insulin was increased from 3 +/- 1 to 15 +/- 2 micro U/ml, and portal insulin was increased from 16 +/- 3 to 33 +/- 3 micro U/ml in PePo after t = 60 min. Endogenous glucose production (R(a)) rose similarly from basal during the first 60 min of exercise in all groups (mean +/- SE of all groups was from 2.2 +/- 0.1 to 6.8 +/- 0.5 mg. kg(-1). min(-1)). The increase in R(a) was sustained for the remainder of the exercise period in C. R(a) was suppressed by approximately 40%, but only after 60 min of hyperinsulinemia, and by approximately 20% after 90 min of hyperinsulinemia in Pe. In contrast, the addition of portal venous hyperinsulinemia caused approximately 90% suppression of R(a) within 20 min and for the remainder of the experiment in PePo. Measurements of net hepatic glucose output were similar to R(a) responses in all groups. Arterial free fatty acids (FFAs), a stimulus of R(a), were increased to 1,255 +/- 258 micro mol/l in C but were only 459 +/- 67 and 312 +/- 42 micro mol/l in Pe and PePo, respectively, by 150 min of exercise. Thus, during exercise, the exquisite sensitivity of R(a) to hyperinsulinemia is due entirely to portal venous hyperinsulinemia during the first 60 min, after which peripheral hyperinsulinemia may control approximately 20-40%, possibly as a result of inhibition of the exercise-induced increase in FFA. PMID- 14747278 TI - Strength training increases insulin-mediated glucose uptake, GLUT4 content, and insulin signaling in skeletal muscle in patients with type 2 diabetes. AB - Strength training represents an alternative to endurance training for patients with type 2 diabetes. Little is known about the effect on insulin action and key proteins in skeletal muscle, and the necessary volume of strength training is unknown. A total of 10 type 2 diabetic subjects and 7 healthy men (control subjects) strength-trained one leg three times per week for 6 weeks while the other leg remained untrained. Each session lasted no more than 30 min. After strength training, muscle biopsies were obtained, and an isoglycemic hyperinsulinemic clamp combined with arterio-femoral venous catheterization of both legs was carried out. In general, qualitatively similar responses were obtained in both groups. During the clamp, leg blood flow was higher (P < 0.05) in trained versus untrained legs, but despite this, arterio-venous extraction glucose did not decrease in trained legs. Thus, leg glucose clearance was increased in trained legs (P < 0.05) and more than explained by increases in muscle mass. Strength training increased protein content of GLUT4, insulin receptor, protein kinase B-alpha/beta, glycogen synthase (GS), and GS total activity. In conclusion, we found that strength training for 30 min three times per week increases insulin action in skeletal muscle in both groups. The adaptation is attributable to local contraction-mediated mechanisms involving key proteins in the insulin signaling cascade. PMID- 14747279 TI - Hexokinase II overexpression improves exercise-stimulated but not insulin stimulated muscle glucose uptake in high-fat-fed C57BL/6J mice. AB - The aim of the present study was to determine the specific sites of impairment to muscle glucose uptake (MGU) in the insulin-resistant high-fat-fed, conscious C57BL/6J mouse. Wild type (WT) and hexokinase II overexpressing (HK(Tg)) mice were fed either a standard diet or high-fat diet and studied at 4 months of age. A carotid artery and jugular veins had catheters chronically implanted for sampling and infusions, respectively, and mice were allowed to recovery for at least 5 days. Mice were fasted for 5 h and underwent a hyperinsulinemic euglycemic clamp or saline infusion for 120 min. Separate groups of mice were studied during 30-min sedentary or treadmill exercise periods. A bolus of 2 deoxy[(3)H]glucose was administered 25 min before the end of each study for determination of R(g), an index of tissue-specific glucose uptake. Fasting blood glucose was increased in high-fat compared with standard diet-fed WT (194 +/- 4 vs. 171 +/- 4 mg/dl) but not HK(Tg) (179 +/- 5 vs. 171 +/- 3 mg/dl) mice. High fat feeding created hyperinsulinemia in both WT and HK(Tg) mice (58 +/- 8 and 77 +/- 15 micro U/ml) compared with standard diet-fed mice (21 +/- 2 and 20 +/- 1 micro U/ml). R(g) was not affected by genotype or diet during either saline infusion or sedentary conditions. HK II overexpression augmented insulin stimulated R(g) in standard diet-fed but not high-fat-fed mice. Exercise stimulated R(g) was impaired by high-fat feeding in WT mice, but this impairment was largely rectified in HK(Tg) mice. In conclusion, high-fat feeding impairs both insulin- and exercise-stimulated MGU, but only exercise-stimulated MGU was corrected by HK II overexpression. PMID- 14747280 TI - Effect of moderate exercise training on peripheral glucose effectiveness, insulin sensitivity, and endogenous glucose production in healthy humans estimated by a two-compartment-labeled minimal model. AB - For examining the effects of moderate exercise training on peripheral glucose effectiveness (S(g)(2)*), insulin sensitivity (S(i)(2)*), and endogenous glucose production (EGP), seven men and one woman (24.8 +/- 1.8 years) participated in cycle ergometer training at lactate threshold intensity for 60 min/day, 5 days/week for 12 weeks. Stable-labeled frequently sampled intravenous glucose tolerance tests were performed before and 16 h and 1 week after the last training session. S(g)(2)* (pre 0.71 +/- 0.03 x 10(-2), 16 h 0.85 +/- 0.02 x 10(-2) dl. kg(-1). min(-1)) and S(i)(2)* (pre 12.6 +/- 2.6 x 10(-4), 16 h 19.7 +/- 3.3 x 10( 4) dl. kg(-1). min(-1). [ micro U/ml](-1)), analyzed using the two-compartment minimal model, were significantly elevated 16 h after the last training session. The elevated S(g)(2)* remained higher despite the cessation of exercise training for 1 week (1.00 +/- 0.03 x 10(-2) dl. kg(-1). min(-1)). EGP was suppressed within 20 min after glucose bolus, and the suppression of EGP was followed by their overshoot. The time course of EGP during the intravenous glucose tolerance test remained similar after the training period. In conclusion, moderate exercise training at lactate threshold improves not only peripheral insulin sensitivity but also peripheral glucose effectiveness with no change in the effect of glucose and/or insulin to suppress EGP in healthy humans. PMID- 14747281 TI - Sterol regulatory element-binding protein-1 mediates the effect of insulin on hexokinase II gene expression in human muscle cells. AB - Insulin upregulates hexokinase II (HKII) expression in skeletal muscle, and this effect is altered in type 2 diabetic patients. This study was conducted to identify the transcription factors that mediate the effect of insulin on HKII gene expression in human muscle. We have cloned the promoter region of the HKII gene and investigated its regulation in a primary culture of human skeletal muscle cells. We defined a region (-369/-270) that conferred the transcriptional response to insulin. This region contains a sterol regulatory element (SRE) that interacted with the recombinant active form of SRE binding protein-1c (SREBP-1c) in electrophoretic mobility shift assays, and, using chromatin immunoprecipitation assay, we showed that endogenous SREBP-1 interacted directly with the promoter region of the HKII gene in human muscle cells. Mutation of the SRE sequence completely suppressed the response of the promoter to insulin stimulation. Finally, overexpression of the rodent mature form of SREBP-1c (adipocyte determination and differentiation factor-1 [ADD1]-403) was able to reproduce insulin action, whereas a dominant-negative form (ADD1-403R) prevented the effect of insulin on HKII promoter constructs. These results demonstrate that SREBP-1c is involved in the effect of insulin on HKII gene transcription and indicate that it is one of the mediators of insulin action on gene expression in human skeletal muscle. PMID- 14747282 TI - Ca2+ and AMPK both mediate stimulation of glucose transport by muscle contractions. AB - It is now generally accepted that activation of AMP-activated protein kinase (AMPK) is involved in the stimulation of glucose transport by muscle contractions. However, earlier studies provided evidence that increases in cytosolic Ca(2+) mediate the effect of muscle contractions on glucose transport. The purpose of this study was to test the hypothesis that both the increase in cytosolic Ca(2+) and the activation of AMPK are involved in the stimulation of glucose transport by muscle contractions. Caffeine causes release of Ca(2+) from the sarcoplasmic reticulum. Incubation of rat epitrochlearis muscles with a concentration of caffeine that raises cytosolic Ca(2+) to levels too low to cause contraction resulted in an approximate threefold increase in glucose transport. Caffeine treatment also resulted in increased phosphorylation of calmodulin dependent protein kinase (CAMK)-II in epitrochlearis muscle. The stimulation of glucose transport by caffeine was blocked by the Ca(2+)-CAMK inhibitors KN62 and KN93. Activation of AMPK with 5-aminoimidazole-4-carboxamide ribonucleoside (AICAR) also resulted in an approximate threefold increase in glucose transport in the epitrochlearis. The increases in glucose transport induced by AICAR and caffeine were additive, and their combined effect was not significantly different from that induced by maximally effective contractile activity. KN62 and KN93 caused an approximately 50% inhibition of the stimulation of glucose transport by contractile activity. Our results provide evidence that both Ca(2+) and AMPK are involved in the stimulation of glucose transport by muscle contractions. They also suggest that the stimulation of glucose transport by Ca(2+) involves activation of CAMK. PMID- 14747283 TI - Prevention of obesity and insulin resistance in mice lacking plasminogen activator inhibitor 1. AB - Increased plasminogen activator inhibitor 1 (PAI-1) has been linked to not only thrombosis and fibrosis but also to obesity and insulin resistance. Increased PAI 1 levels have been presumed to be consequent to obesity. We investigated the interrelationships of PAI-1, obesity, and insulin resistance in a high-fat/high carbohydrate (HF) diet-induced obesity model in wild-type (WT) and PAI-1 deficient mice (PAI-1(-/-)). Obesity and insulin resistance developing in WT mice on an HF diet were completely prevented in mice lacking PAI-1. PAI-1(-/-) mice on an HF diet had increased resting metabolic rates and total energy expenditure compared with WT mice, along with a marked increase in uncoupling protein 3 mRNA expression in skeletal muscle, likely mechanisms contributing to the prevention of obesity. In addition, insulin sensitivity was enhanced significantly in PAI-1( /-) mice on an HF diet, as shown by euglycemic-hyperinsulinemic clamp studies. Peroxisome proliferator-activated receptor (PPAR)-gamma and adiponectin mRNA, key control molecules in lipid metabolism and insulin sensitivity, were maintained in response to an HF diet in white adipose tissue in PAI-1(-/-) mice, contrasting with downregulation in WT mice. This maintenance of PPAR-gamma and adiponectin may also contribute to the observed maintenance of body weight and insulin sensitivity in PAI-1(-/-) mice. Treatment in WT mice on an HF diet with the angiotensin type 1 receptor antagonist to downregulate PAI-1 indeed inhibited PAI 1 increases and ameliorated diet-induced obesity, hyperglycemia, and hyperinsulinemia. PAI-1 deficiency also enhanced basal and insulin-stimulated glucose uptake in adipose cells in vitro. Our data suggest that PAI-1 may not merely increase in response to obesity and insulin resistance, but may have a direct causal role in obesity and insulin resistance. Inhibition of PAI-1 might provide a novel anti-obesity and anti-insulin resistance treatment. PMID- 14747285 TI - Proinsulin expression by Hassall's corpuscles in the mouse thymus. AB - The thymus expresses proinsulin, among many other tissue-specific antigens, and the inheritance of genetically determined low thymic proinsulin expression has been associated with impaired proinsulin-specific autoreactive T-cell tolerance and type 1 diabetes susceptibility. The cellular and molecular biology of proinsulin expression in the thymus remains unknown, and contradictory reports exist regarding the identity of proinsulin-producing cells. Using knock-in mice expressing beta-galactosidase (beta-Gal) under the control of an endogenous insulin promoter, we found that thymic proinsulin and beta-Gal transcripts were detectable at high levels in purified thymic epithelial cells. Immunohistochemical analysis of beta-Gal activity showed that most proinsulin expression can be accounted for by rare medullary epithelial cells of the Hassall's corpuscles. Moreover, flow cytometry analyses of beta-Gal-positive cells showed that only 1-3% of all epithelial cells express proinsulin, and this technique will now provide us with a method for isolating the proinsulin producing cells in mouse thymus. PMID- 14747284 TI - Leptin impairs insulin signaling in rat adipocytes. AB - Leptin modulates glucose homeostasis by acting as an insulin-sensitizing factor in most insulin target tissues. Nevertheless, insulin-dependent glucose uptake in white adipose tissue decreases after in vivo treatment with leptin. Moreover, elevated leptin concentrations inhibit insulin metabolic effects in adipocytes. Here we studied both, direct and centrally mediated effects of leptin on insulin signaling in rat adipocytes. Adipocyte incubation with low leptin concentrations did not modify the insulin stimulation of mitogen-activated protein kinase (MAPK). However, at elevated concentrations, leptin impaired insulin-stimulated MAPK activity, glycogen synthase kinase (GSK)3beta phosphorylation, and insulin receptor tyrosine phosphorylation without altering vanadate stimulation. An increase of suppressor of cytokine signaling-3 protein was also observed. Central administration of leptin decreased insulin effects on adipocyte MAPK and GSK3beta phosphorylation. In insulin-resistant aged rats with hyperleptinemia and central leptin resistance, insulin poorly stimulated MAPK and central leptin infusion did not further deteriorate adipocyte insulin responsiveness. Food restriction increased MAPK stimulation by insulin and restored the ability of centrally infused leptin to attenuate adipocyte insulin signaling in aged rats. We conclude that leptin can modulate, in an inhibitory manner, adipocyte insulin signaling by two different ways: as an autocrine signal and, indirectly, through neuroendocrine pathways. These mechanisms may be of relevance in situations of hyperleptinemia, such as aging and/or obesity. PMID- 14747286 TI - Exercise induces hypoglycemia in rats with islet transplantation. AB - Recently, islet transplantation in patients with type 1 diabetes has had greater success than in the past, but the important question of whether the kinetics of islet secretion are able to accommodate the metabolic demands of special conditions such as exercise remains unanswered. Syngeneic rat islets (4,000 islet equivalents/rat) were transplanted into the liver, kidney, and peritoneal cavity (encapsulated or nonencapsulated) of rats with streptozocin-induced diabetes. Normoglycemic transplanted rats and age-matched controls were subjected to 30 min of moderate exercise on a treadmill 5 weeks after transplantation. Although control rats maintained near normoglycemia during and after exercise, the rats with islet transplants had significantly lower blood glucose levels. For the rats with islets in the liver, increased C-peptide levels were found at 30 min (790 +/ 125 and 1,450 +/- 250 pmol/l at 0 and 30 min, respectively; P < 0.01), whereas a decrease was found in controls and in rats with islets transplanted into the peritoneal cavity or under the kidney capsule. Moreover, increased glucagon levels were found after exercise in the rats with islets transplanted into the liver (62 +/- 6, 165 +/- 29, 155 +/- 27, and 97 +/- 13 pg/ml at 0, 30, 60, and 90 min, respectively; P < 0.05), whereas no changes in glucagon levels were observed in controls. In conclusion, moderate exercise caused hypoglycemia in rats with islet transplants in different sites including liver, kidney, and peritoneal cavity. C-peptide and glucagon responses to exercise were very different in rats with transplanted islets compared with controls. This islet dysfunction led to exercise-induced hypoglycemia. PMID- 14747287 TI - Gene expression profiles define a key checkpoint for type 1 diabetes in NOD mice. AB - cDNA microarrays with >11,000 cDNA clones from an NOD spleen cDNA library were used to identify temporal gene expression changes in NOD mice (1-10 weeks), which spontaneously develop type 1 diabetes, and changes between NOD and NOD congenic mice (NOD.Idd3/Idd10 and NOD.B10Sn-H2(b)), which have near zero incidence of insulitis and diabetes. The expression profiles identified two distinct groups of mice corresponding to an immature (1-4 weeks) and mature (6-10 weeks) state. The rapid switch of gene expression occurring around 5 weeks of age defines a key immunological checkpoint. Sixty-two known genes are upregulated, and 18 are downregulated at this checkpoint in the NOD. The expression profiles are consistent with increased antibody production, antigen presentation, and cell proliferation associated with an active autoimmune response. Seven of these genes map to confirmed diabetes susceptibility regions. Of these seven, three are excellent candidate genes not previously implicated in type 1 diabetes. Ten genes are differentially expressed between the NOD and congenic NOD at the immature stage (Hspa8, Hif1a, and several involved in cellular functions), while the other 70 genes exhibit expression differences during the mature (6-10 week) stage, suggesting that the expression differences of a small number of genes before onset of insulitis determine the disease progression. PMID- 14747288 TI - Mixed hematopoietic chimerism allows cure of autoimmune diabetes through allogeneic tolerance and reversal of autoimmunity. AB - Bone marrow transplantation from diabetes-resistant strains with complete replacement of the recipient immune system by the allogeneic donor has led to tolerance to donor islets and cure of diabetes in a mouse model of type 1 diabetes. However, the ability to tolerize host T-cells of diabetic NOD mice is unknown. We demonstrate that nonmyeloablative conditioning achieves mixed hematopoietic chimerism across major histocompatibility complex (MHC) barriers in spontaneously diabetic NOD mice. This conditioning preserves alloreactive and autoreactive diabetogenic host NOD T-cells, but when mixed chimerism was established, diabetic NOD mice accepted donor-type allogeneic islet grafts and were cured of diabetes, despite a significant recipient T-cell contribution. Furthermore, induction of mixed chimerism permitted acceptance of NOD islet grafts, demonstrating reversal of autoimmunity. Allogeneic bone marrow transplantation was critical for tolerization of diabetogenic and alloreactive host T-cells. Thus, mixed hematopoietic chimerism induces tolerance to donor islets and reverses established autoimmunity in diabetic NOD mice. PMID- 14747289 TI - Stratification of type 1 diabetes risk on the basis of islet autoantibody characteristics. AB - Family history of type 1 diabetes and autoantibodies to the islet antigens insulin (IAA), glutamate decarboxylase (GADA), and the protein tyrosine phosphatase-like protein IA-2 (IA-2A) are strong predictors of type 1 diabetes, but the rate of progression to diabetes in multiple islet autoantibody-positive relatives varies widely. We asked whether detailed characterization of islet autoantibodies that included determination of titer, epitope specificity, and IgG subclass would improve diabetes prediction in a large cohort of autoantibody positive relatives. The study shows a strong association between risk and high titer, broad antibody responses to IA-2 and insulin. The highest risks were associated with high-titer IA-2A and IAA, IgG2, IgG3, and/or IgG4 subclass of IA 2A and IAA, and antibodies to the IA-2-related molecule IA-2beta. Using models based on these antibody characteristics, autoantibody-positive relatives can be classified into groups with risks of diabetes ranging from 7 to 89% within 5 years. PMID- 14747290 TI - Role for plasma membrane-related Ca2+-ATPase-1 (ATP2C1) in pancreatic beta-cell Ca2+ homeostasis revealed by RNA silencing. AB - Changes in intracellular Ca(2+) concentration play a key role in the regulation of insulin secretion by glucose and other secretagogues. Here, we explore the importance of the secretory pathway Ca(2+)-ATPase, plasma membrane-related Ca(2+) ATPase-1 (PMR1; human orthologue ATP2C1) in intracellular Ca(2+) homeostasis in pancreatic islet beta-cells. Endogenous PMR1 mRNA and protein were detected in both isolated rat islets and beta-cell-derived lines (MIN6 and INS1). Subcellular fractionation of the cell lines revealed PMR1 immunoreactivity in both microsomal and dense-core secretory vesicle-enriched fractions. Correspondingly, depletion of cellular PMR1 with small interfering RNAs inhibited Ca(2+) uptake into the endoplasmic reticulum and secretory vesicles by approximately 20%, as assessed using organelle-targeted aequorins in permeabilized INS1 cells. In intact cells, PMR1 depletion markedly enhanced flux though L-type Ca(2+) channels and augmented glucose-stimulated, but not basal, insulin secretion. Whereas average cytosolic [Ca(2+)] increases in response to 30.0 mmol/l glucose were unaffected by PMR1 depletion, [Ca(2+)] oscillation shape, duration, and decay rate in response to glucose plus tetraethylammonium were modified in PMR1-depleted single cells, imaged using fluo-3-acetoxymethylester. PMR1 thus plays an important role, which is at least partially nonoverlapping with that of sarco(endo-)plasmic reticulum Ca(2+)-ATPases, in the control of beta-cell Ca(2+) homeostasis and insulin secretion. PMID- 14747291 TI - Regulation of ATP/ADP in pancreatic islets. AB - ATP and ADP levels are critical regulators of glucose-stimulated insulin secretion. In many aerobic cell types, the phosphorylation potential (ATP/ADP/P(i)) is controlled by sensing mechanisms inherent in mitochondrial metabolism that feed back and induce compensatory changes in electron transport. To determine whether such regulation may contribute to stimulus-secretion coupling in islet cells, we used a recently developed flow culture system to continuously and noninvasively measure cytochrome c redox state and oxygen consumption as indexes of electron transport in perifused isolated rat islets. Increasing substrate availability by increasing glucose increased cytochrome c reduction and oxygen consumption, whereas increasing metabolic demand with glibenclamide increased oxygen consumption but not cytochrome c reduction. The data were analyzed using a kinetic model of the dual control of electron transport and oxygen consumption by substrate availability and energy demand, and ATP/ADP/P(i) was estimated as a function of time. ATP/ADP/P(i) increased in response to glucose and decreased in response to glibenclamide, consistent with what is known about the effects of these agents on energy state. Therefore, a simple model representing the hypothesized role of mitochondrial coupling in governing phosphorylation potential correctly predicted the directional changes in ATP/ADP/P(i). Thus, the data support the notion that mitochondrial-coupling mechanisms, by virtue of their role in establishing ATP and ADP levels, may play a role in mediating nutrient-stimulated insulin secretion. Our results also offer a new method for continuous noninvasive measures of islet cell phosphorylation potential, a critical metabolic variable that controls insulin secretion by ATP sensitive K(+)-dependent and -independent mechanisms. PMID- 14747292 TI - Reduction in glucagon receptor expression by an antisense oligonucleotide ameliorates diabetic syndrome in db/db mice. AB - Excess glucagon levels contribute to the hyperglycemia associated with type 2 diabetes. Reducing glucagon receptor expression may thus ameliorate the consequences of hyperglucagonemia and improve blood glucose control in diabetic patients. This study describes the antidiabetic effects of a specific glucagon receptor antisense oligonucleotide (GR-ASO) in db/db mice. The ability of GR-ASOs to inhibit glucagon receptor mRNA expression was demonstrated in primary mouse hepatocytes by quantitative real-time RT-PCR. Intraperitoneal administration of GR-ASO at a dosage of 25 mg/kg twice a week in db/db mice for 3 weeks resulted in 1) decreased glucagon receptor mRNA expression in liver; 2) decreased glucagon stimulated cAMP production in hepatocytes isolated from GR-ASO-treated db/db mice; 3) significantly reduced blood levels of glucose, triglyceride, and free fatty acids; 4) improved glucose tolerance; and 5) a diminished hyperglycemic response to glucagon challenge. Neither lean nor db/db mice treated with GR-ASO exhibited hypoglycemia. Suppression of GR expression was also associated with increased ( approximately 10-fold) levels of plasma glucagon. No changes were observed in pancreatic islet cytoarchitecture, islet size, or alpha-cell number. However, alpha-cell glucagon levels were increased significantly. Our studies support the concept that antagonism of glucagon receptors could be an effective approach for controlling blood glucose in diabetes. PMID- 14747293 TI - Effect of recurrent hypoglycemia on spatial cognition and cognitive metabolism in normal and diabetic rats. AB - The effects of recurrent hypoglycemia (RH) on cognition in human subjects remain controversial, perhaps in part due to difficulty in completely controlling previous hypoglycemic history. We used a model of RH in nondiabetic and diabetic rats to examine the effects of short-term (3 h daily for 3 days) RH on subsequent hippocampally dependent spatial memory, tested either at euglycemia or under acute hypoglycemia. Hippocampal metabolism was simultaneously measured using microdialysis. Antecedent RH improved task performance (79 +/- 2% alternation in nondiabetic RH animals vs. 63 +/- 3% in controls; P < 0.001) at euglycemia, accompanied by reversal of the task-associated dip (20 +/- 1% below baseline) in hippocampal extracellular fluid (ECF) glucose seen in control animals. RH rats also had a larger rise in hippocampal ECF glucose, after intraperitoneal glucose injection, than did controls. However, RH animals tested at acute hypoglycemia ( approximately 2.8 mmol/l) performed significantly worse than control animals. Results were similar in diabetic and nondiabetic rats. Our data suggest that RH causes improvement in subsequent cognitive performance at euglycemia, accompanied by alterations in cognitive metabolism. When glucose availability is limited, complex cognitive functioning seems to be adversely effected in RH animals, perhaps to better maintain and preserve basic brain functions. PMID- 14747294 TI - Insulin secretion in type 1 diabetes. AB - Type 1 diabetes, a chronic autoimmune disease, causes destruction of insulin producing beta-cells over a period of years. Although many markers of the autoimmune process have been described, none can convincingly predict the rate of disease progression. Moreover, there is relatively little information about changes in insulin secretion in individuals with type 1 diabetes over time. Previous studies document C-peptide at a limited number of time points, often after a nonphysiologic stimulus, and under non-steady-state conditions. Such methods do not provide qualitative information and may not reflect physiologic responses. We have studied qualitative and quantitative insulin secretion to a 4 h mixed meal in 41 patients with newly diagnosed type 1 diabetes and followed the course of this response for 24 months in 20 patients. Newly diagnosed diabetic patients had an average total insulin secretion in response to a mixed meal that was 52% of that in nondiabetic control subjects, considerably higher than has been described previously. In diabetic patients there was a decline of beta-cell function at an average rate of 756 +/- 132 pmol/month to a final value of 28 +/- 8.4% of initial levels after 2 years. There was a significant correlation between the total insulin secretory response and control of glucose, measured by HbA(1c) (P = 0.003). Two persistent patterns of insulin response were seen depending on the peak insulin response following the oral meal. Patients with an early insulin response (i.e., within the first 45 min after ingestion) to a mixed meal, which was also seen in 37 of 38 nondiabetic control subjects, had a significantly accelerated loss of insulin secretion, as compared with those in whom the insulin response occurred after this time (P < 0.05), and significantly greater insulin secretory responses at 18 and 24 months (P < 0.02). These results, which are the first qualitative studies of insulin secretion in type 1 diabetes, indicate that the physiologic metabolic response is greater at diagnosis than has previously been appreciated, and that the qualitative insulin secretory response is an important determinant of the rate of metabolic decompensation from autoimmune destruction. PMID- 14747295 TI - Electrocardiographic repolarization complexity and abnormality predict all-cause and cardiovascular mortality in diabetes: the strong heart study. AB - Type 2 diabetes is associated with increased risk of cardiovascular (CV) and all cause mortality. Although electrocardiographic measures of repolarization abnormality and complexity stratify risk in the general population, their prognostic value in diabetes has not been well characterized. Digital electrocardiogram (ECG) readings were acquired for 994 American Indians with type 2 diabetes. ST segment depression (STD) >/=50 micro V and rate-corrected QT interval (QTc) >460 ms were examined as measures of repolarization abnormality. The principal component analysis (PCA) of the ratio of the second to first eigenvalues of the T-wave vector (PCA ratio) (>32.0% in women and >24.6% in men) was examined as a measure of repolarization complexity on the ECG. After a mean follow-up of 4.7 +/- 1.0 years, there were 56 CV deaths and 155 deaths from all causes. In univariate analyses, STD, QTc, and the PCA ratio predicted CV and all cause mortality. After multivariate adjustment for age, sex, and other risk factors, STD (hazard ratio 3.68, 95% CI 1.70-7.96) and PCA ratio (2.61, 1.33 5.13) remained predictive of CV mortality and both STD (2.36, 1.38-4.02) and QTc (2.03, 1.32-3.12) predicted all-cause mortality. Computerized ECG measures of repolarization abnormality and complexity predict CV and all-cause mortality in type 2 diabetes, supporting their use to identify high-risk individuals with diabetes. PMID- 14747296 TI - Decrease in glucose-stimulated insulin secretion with aging is independent of insulin action. AB - While the incidence of diabetes increases with age, a decrease in beta-cell function independent of age-related insulin resistance has not been conclusively determined. We studied insulin secretion (by hyperglycemic clamp) in 3-, 9-, and 20-month-old chronically catheterized, awake, Sprague Dawley (SD) rats (n = 78). Insulin action was modulated in a group of old rats by caloric restriction (CR) or by surgical removal of visceral fat (VF-). During the first 2 h of the clamp (11 mmol/l glucose), insulin secretion and insulin resistance (S(i hyper clamp)) demonstrated the characteristic hyperbolic relationship. However, after hyperglycemia for an additional 2 h, the ability to maintain insulin secretion, commensurate with the degree of insulin resistance, was decreased in all aging rats (P < 0.05). Increasing plasma glucose levels to 18 mmol/l glucose, after clamp at 11 mmol/l, increased insulin secretion by approximately threefold in young rats, but failed to induce similar magnitude of response in the aging rats ( approximately 50%). However, elevation of plasma free fatty acid (FFA) levels by twofold (by intralipid infusion during 11 mmol/l glucose clamp) resulted in a robust, approximate twofold response in both young and old rats. Thus, prolonged stimulation by hyperglycemia unveiled a functional defect in insulin secretion with aging. This age-related defect is independent of insulin action and is specific to glucose and not FFAs. We suggest that prolonged hyperglycemic stimulation can be a tool to identify functional defects in insulin secretion, particularly in the context of the hyperbolic relationship with insulin action, in elderly subjects or those at risk for type 2 diabetes. PMID- 14747297 TI - Insulin sensitivity of muscle capillary recruitment in vivo. AB - We have reported that insulin exerts two vascular actions in muscle; it both increases blood flow and recruits capillaries. In parallel hyperinsulinemic euglycemic clamp studies, we compared the insulin dose response of muscle microvascular recruitment and femoral blood flow as well as hindleg glucose uptake in fed, hooded Wistar and fasted Sprague-Dawley rats. Using insulin doses between 0 and 30 mU(-1). min(-1). kg(-1), we measured microvascular recruitment at 2 h by 1-methylxanthine (1-MX) metabolism or contrast-enhanced ultrasound (CEU), and muscle glucose uptake was measured by either arteriovenous differences or using 2-deoxyglucose. We also examined the time course for reversal of microvascular recruitment following cessation of a 3 mU. min(-1). kg(-1) insulin infusion. In both groups, whether measured by 1-MX metabolism or CEU, microvascular recruitment was fully activated by physiologic hyperinsulinemia and occurred at lower insulin concentrations than those that stimulated glucose uptake or hindleg total blood flow. The latter processes were insulin dose dependent throughout the entire dose range studied. Upon stopping the insulin infusion, increases in microvascular volume persisted for 15-30 min after insulin concentrations returned to basal levels. We conclude that the precapillary arterioles that regulate microvascular recruitment are more insulin sensitive than resistance arterioles that regulate total flow. PMID- 14747298 TI - Absence of inducible nitric oxide synthase reduces myocardial damage during ischemia reperfusion in streptozotocin-induced hyperglycemic mice. AB - We investigated the role of inducible nitric oxide synthase (iNOS) on ischemic myocardial damage and angiogenic process in genetically deficient iNOS (iNOS(-/ )) mice and wild-type littermates (iNOS(+/+)), with and without streptozotocin induced (70 mg/kg intravenously) diabetes. After ischemia (25 min) and reperfusion (120 min), both iNOS(+/+) and iNOS(-/-) diabetic mice (blood glucose 22 mmol/l) had myocardial infarct size greater than their respective nondiabetic littermates (P < 0.01). Myocardial infarct size (P < 0.05), apoptotic index (P < 0.005), and tissue levels of tumor necrosis factor (P < 0.01), interleukin-6 (P < 0.01), and interleukin-18 (P < 0.01) were higher in nondiabetic iNOS(-/-) mice compared with nondiabetic iNOS(+/+) mice. As compared with diabetic iNOS(-/-) mice, diabetic iNOS(+/+) mice showed a greater infarct size (P < 0.01) associated with the highest tissue levels of nitrotyrosine and proinflammatory cytokines, as well as apoptosis. The beneficial role of iNOS in modulating defensive responses against ischemia/reperfusion injury seems to be abolished in diabetic mice. PMID- 14747299 TI - Diabetes increases formation of advanced glycation end products on Sarco(endo)plasmic reticulum Ca2+-ATPase. AB - Prolongation of relaxation is a hallmark of diabetic cardiomyopathy. Most studies attribute this defect to decreases in sarco(endo)plasmic reticulum Ca(2+)-ATPase (SERCA2a) expression and SERCA2a-to-phospholamban (PLB) ratio. Since its turnover rate is slow, SERCA2a is susceptible to posttranslational modifications during diabetes. These modifications could in turn compromise conformational rearrangements needed to translocate calcium ions, also leading to a decrease in SERCA2a activity. In the present study one such modification was investigated, namely advanced glycation end products (AGEs). Hearts from 8-week streptozotocin induced diabetic (8D) rats showed typical slowing in relaxation, confirming cardiomyopathy. Hearts from 8D animals also expressed lower levels of SERCA2a protein and higher levels of PLB. Analysis of matrix-assisted laser desorption/ionization time-of-flight mass data files from trypsin-digested SERCA2a revealed several cytosolic SERCA2a peptides from 8D modified by single noncrosslinking AGEs. Crosslinked AGEs were also found. Lysine residues within actuator and phosphorylation domains were cross-linked to arginine residues within the nucleotide binding domain via pentosidine AGEs. Two weeks of insulin treatment initiated after 6 weeks of diabetes attenuated these changes. These data demonstrate for the first time that AGEs are formed on SERCA2a during diabetes, suggesting a novel mechanism by which cardiac relaxation can be slowed during diabetes. PMID- 14747300 TI - Increased risk of type 2 diabetes in Alzheimer disease. AB - Alzheimer disease and type 2 diabetes are characterized by increased prevalence with aging, a genetic predisposition, and comparable pathological features in the islet and brain (amyloid derived from amyloid beta protein in the brain in Alzheimer disease and islet amyloid derived from islet amyloid polypeptide in the pancreas in type 2 diabetes). Evidence is growing to link precursors of amyloid deposition in the brain and pancreas with the pathogenesis of Alzheimer disease and type 2 diabetes, respectively. Given these similarities, we questioned whether there may be a common underlying mechanism predisposing to islet and cerebral amyloid. To address this, we first examined the prevalence of type 2 diabetes in a community-based controlled study, the Mayo Clinic Alzheimer Disease Patient Registry (ADPR), which follows patients with Alzheimer disease versus control subjects without Alzheimer disease. In addition to this clinical study, we performed a pathological study of autopsy cases from this same community to determine whether there is an increased prevalence of islet amyloid in patients with Alzheimer disease and increased prevalence of cerebral amyloid in patients with type 2 diabetes. Patients who were enrolled in the ADPR (Alzheimer disease n = 100, non-Alzheimer disease control subjects n = 138) were classified according to fasting glucose concentration (FPG) as nondiabetic (FPG <110 mg/dl), impaired fasting glucose (IFG, FPG 110-125 mg/dl), and type 2 diabetes (FPG >126 mg/dl). The mean slope of FPG over 10 years in each case was also compared between Alzheimer disease and non-Alzheimer disease control subjects. Pancreas and brain were examined from autopsy specimens obtained from 105 humans (first, 28 cases of Alzheimer disease disease vs. 21 non-Alzheimer disease control subjects and, second, 35 subjects with type 2 diabetes vs. 21 non-type 2 diabetes control subjects) for the presence of islet and brain amyloid. Both type 2 diabetes (35% vs. 18%; P < 0.05) and IFG (46% vs. 24%; P < 0.01) were more prevalent in Alzheimer disease versus non-Alzheimer disease control subjects, so 81% of cases of Alzheimer disease had either type 2 diabetes or IFG. The slope of increase of FPG with age over 10 years was also greater in Alzheimer disease than non Alzheimer disease control subjects (P < 0.01). Islet amyloid was more frequent (P < 0.05) and extensive (P < 0.05) in patients with Alzheimer disease than in non Alzheimer disease control subjects. However, diffuse and neuritic plaques were not more common in type 2 diabetes than in control subjects. In cases of type 2 diabetes when they were present, the duration of type 2 diabetes correlated with the density of diffuse (P < 0.001) and neuritic plaques (P < 0.01). In this community cohort from southeast Minnesota, type 2 diabetes and IFG are more common in patients with Alzheimer disease than in control subjects, as is the pathological hallmark of type 2 diabetes, islet amyloid. However, there was no increase in brain plaque formation in cases of type 2 diabetes, although when it was present, it correlated in extent with duration of diabetes. These data support the hypothesis that patients with Alzheimer disease are more vulnerable to type 2 diabetes and the possibility of linkage between the processes responsible for loss of brain cells and beta-cells in these diseases. PMID- 14747301 TI - Uncoupling protein 2 promoter polymorphism -866G/A affects its expression in beta cells and modulates clinical profiles of Japanese type 2 diabetic patients. AB - Common uncoupling protein 2 (UCP2) promoter polymorphism -866G/A is reported to be associated with its expression in adipose tissue and the risk of obesity in Caucasians. On the other hand, several studies suggested that UCP2 expression in beta-cells is an important determinant of insulin secretion. In the Japanese population, morbid obesity is very rare, and insulin secretion capacity is relatively low as compared with Caucasians. Because UCP2 would link to insulin secretion and obesity, it might explain this ethnic difference. Here, we report that the UCP2 promoter with the A allele showed higher promoter activity in the INS-1 beta-cell line. The frequency of the A allele is higher in our Japanese study than that in Caucasians. Type 2 diabetic patients with the A allele need insulin therapy earlier and showed higher frequency of insulin treatment. Moreover glucose-induced early insulin secretion is significantly lower in patients with the A allele. However, there was no difference in allele frequency between obese and lean type 2 diabetic patients. In conclusion, UCP2 promoter polymorphism -866G/A does not affect obesity in Japanese type 2 diabetic patients but affects its transcription in beta-cells and modulates glucose-induced insulin secretion and eventually insulin requirement in Japanese type 2 diabetic patients. Higher A allele frequency in the Japanese population might partly explain the ethnic difference of insulin secretion capacity. PMID- 14747302 TI - Examination of candidate chromosomal regions for type 2 diabetes reveals a susceptibility locus on human chromosome 8p23.1. AB - In a panel of large Caucasian pedigrees, we genotyped markers in eight chromosomal regions previously reported as supporting linkage with type 2 diabetes. We previously reported significant linkage on chromosome 20q (maximum logarithm of odds score [MLS] = 2.79) in this panel. In the present analysis, candidate regions on 1q, 2q, 3q, 5q, 9q, and 10q yielded little evidence for linkage; a region on 2p (MLS = 1.64, P = 0.01 at position 9.0 cM) gave suggestive evidence of linkage; and a region on 8p (MLS = 3.67, P = 2.8 x 10(-5), at position 7.6 cM) gave significant evidence of linkage. Conditional analyses were performed for both 2p and 8p regions and the region reported on 20q. The MLS for 2p increased from 1.64 to 1.79 (empirical P = 0.142) when conditioned for heterogeneity on 20q. The case was similar for 8p, where the MLS increased from 3.67 to 4.51 (empirical P = 0.023) when conditioned on families without evidence of linkage at 20q. In conclusion, our data support a type 2 diabetes susceptibility locus on chromosome 8p that appears to be independent from other susceptibility loci. Although we were able to replicate linkage in our pedigrees on chromosome 2p, we did not find evidence of linkage for regions on 1q, 2q, 3q, 5q, 9q, or 10q. PMID- 14747303 TI - Linkage and association mapping of a chromosome 1q21-q24 type 2 diabetes susceptibility locus in northern European Caucasians. AB - We have identified a region on chromosome 1q21-q24 that was significantly linked to type 2 diabetes in multiplex families of Northern European ancestry and also in Pima Indians, Amish families, and families from France and England. We sought to narrow and map this locus using a combination of linkage and association approaches by typing microsatellite markers at 1.2 and 0.5 cM densities, respectively, over a region of 37 cM (23.5 Mb). We tested linkage by parametric and nonparametric approaches and association using both case-control and family based methods. In the 40 multiplex families that provided the previous evidence for linkage, the highest parametric, recessive logarithm of odds (LOD) score was 5.29 at marker D1S484 (168.5 cM, 157.5 Mb) without heterogeneity. Nonparametric linkage (NPL) statistics (P = 0.00009), SimWalk2 Statistic A (P = 0.0002), and sib-pair analyses (maximum likelihood score = 6.07) all mapped to the same location. The one LOD CI was narrowed to 156.8-158.9 Mb. Under recessive, two point linkage analysis, adjacent markers D1S2675 (171.5 cM, 158.9 Mb) and D1S1679 (172 cM, 159.1 Mb) showed LOD scores >3.0. Nonparametric analyses revealed a second linkage peak at 180 cM near marker D1S1158 (163.3 Mb, NPL score 3.88, P = 0.0001), which was also supported by case-control (marker D1S194, 178 cM, 162.1 Mb; P = 0.003) and family-based (marker ATA38A05, 179 cM, 162.5 Mb; P = 0.002) association studies. We propose that the replicated linkage findings actually encompass at least two closely spaced regions, with a second susceptibility region located telomeric at 162.5-164.7 Mb. PMID- 14747304 TI - Messenger RNA transcripts of the hepatocyte nuclear factor-1alpha gene containing premature termination codons are subject to nonsense-mediated decay. AB - Mutations in the hepatocyte nuclear factor-1alpha (HNF-1a) gene cause maturity onset diabetes of the young (MODY). Approximately 30% of these mutations generate mRNA transcripts harboring premature termination codons (PTCs). Degradation of such transcripts by the nonsense-mediated decay (NMD) pathway has been reported for many genes. To determine whether PTC mutant transcripts of the HNF-1alpha gene elicit NMD, we have developed a novel quantitative RT-PCR assay. We performed quantification of ectopically expressed mutant transcripts relative to normal transcripts in lymphoblastoid cell lines using a coding single nucleotide polymorphism (cSNP) as a marker. The nonsense mutations R171X, I414G415ATCG- >CCA, and P291fsinsC showed reduced mutant mRNA expression to 40% (P = 0.009), <0.01% (P 360 meV for the tetra-charged pyranine. PMID- 14747307 TI - Diffusion and electrophoretic mobility of single-stranded RNA from molecular dynamics simulations. AB - Hydrodynamic properties of small single-stranded RNA homopolymers with three and six nucleotides in free solution are determined from molecular dynamics simulations in explicit solvent. We find that the electrophoretic mobility increases with increasing RNA length, consistent with experiment. Diffusion coefficients of RNA, corrected for finite-size effects and solvent viscosity, agree well with those estimated from experiments and hydrodynamic calculations. The diffusion coefficients and electrophoretic mobilities satisfy a Nernst Einstein relation in which the effective charge of RNA is reduced by the charge of transiently bound counterions. Fluctuations in the counterion atmosphere are shown to enhance the diffusive spread of RNA molecules drifting along the direction of the external electric field. As a consequence, apparent diffusion coefficients measured by capillary zone electrophoresis can be significantly larger than the actual values at certain experimental conditions. PMID- 14747308 TI - Ionization states of residues in OmpF and mutants: effects of dielectric constant and interactions between residues. AB - To understand ion permeation, one must assign correct ionization states to titratable amino acid residues in protein channels. We report on the effects of physical and methodological assumptions in calculating the protonation states at neutral bulk pH of titratable residues lining the lumen of the native Escherichia coli OmpF channel, and five mutants. We systematically considered a wide range of assumed protein dielectric constants and all plausible combinations of protonation states for electrostatically interacting side chains, and three different levels of accounting for solute shielding: 1), full nonlinear Poisson Boltzmann; 2), linearized Poisson-Boltzmann; and 3), neglect of solute shielding. For this system we found it acceptable to neglect solute shielding, a result we postulate to be generalizable to narrow lumens of other protein channels. For the large majority of residues, the protonation state at neutral bulk pH was found to be independent of the assumed dielectric constant of the protein, and unambiguously determined by the calculation; for native OmpF only Asp-127 has a protonation state that is sensitive to the assumed protein dielectric constant. Our results are significant for understanding two published experimental observations: the structure of the narrow part of the channel, and the ionic selectivity of OmpF mutants. PMID- 14747309 TI - Dynamics of water molecules in the bacteriorhodopsin trimer in explicit lipid/water environment. AB - Protonated networks of internal water molecules appear to be involved in proton transfer in various integral membrane proteins. High-resolution x-ray studies of protein crystals at low temperature deliver mean positions of most internal waters, but only limited information about fluctuations within such H-bonded networks formed by water and residues. The question arises as to how water molecules behave inside and on the surface of a fluctuating membrane protein under more physiological conditions. Therefore, as an example, long-time molecular dynamics simulations of bacteriorhodopsin were performed with explicit membrane/water environment. Based on a recent x-ray model the bacteriorhodopsin trimer was inserted in a fully solvated 16 x 16 1-palmitoyl-2-oleoyl-sn-glycero-3 phosphocholine (POPC)-bilayer patch, resulting in a system of approximately 84,000 atoms. Unrestrained molecular dynamics calculations of 5 ns were performed using the GROMACS package and force field. Mean water densities were computed to describe the anisotropic distribution of internal water molecules. In the whole protein two larger areas of higher water density are identified. They are located between the central proton binding site, the Schiff base, and the extracellular proton release site. Separated by Arg-82 these water clusters could provide a proton release pathway in a Grotthus-like mechanism as indicated by a continuum absorbance change observed during the photocycle by time-resolved Fourier transform infrared spectroscopy. Residues are identified which are H-bonded to the water clusters and are therefore facilitating proton conduction. Their influence on proton transfer via the H-bonded network as indicated by the continuum absorbance change is predicted. This may explain why several site directed mutations alter the proton release kinetics without a direct involvement in proton transfer. PMID- 14747310 TI - Sensitivity, specificity, and the hybridization isotherms of DNA chips. AB - Competitive hybridization, at the surface and in the bulk, lowers the sensitivity of DNA chips. Competitive surface hybridization occurs when different targets can hybridize with the same probe. Competitive bulk hybridization takes place when the targets can hybridize with free complementary chains in the solution. The effects of competitive hybridization on the thermodynamically attainable performance of DNA chips are quantified in terms of the hybridization isotherms of the spots. These relate the equilibrium degree of the hybridization to the bulk composition. The hybridization isotherm emerges as a Langmuir isotherm modified for electrostatic interactions within the probe layer. The sensitivity of the assay in equilibrium is directly related to the slope of the isotherm. A simpler description is possible, in terms of c(50) values specifying the bulk composition corresponding to 50% hybridization at the surface. The effects of competitive hybridization are important for the quantitative analysis of DNA chip results, especially when used to study point mutations. PMID- 14747311 TI - Polyproline II helix conformation in a proline-rich environment: a theoretical study. AB - Interest centers here on whether a polyproline II helix can propagate through adjacent non-proline residues, and on shedding light on recent experimental observations suggesting the presence of significant PP(II) structure in a short alanine-based peptide with no proline in the sequence. For this purpose, we explored the formation of polyproline II helices in proline-rich peptides with the sequences Ac-(Pro)(3)-X-(Pro)(3)-Gly-Tyr-NH(2), with X = Pro (PPP), Ala (PAP), Gln (PQP), Gly (PGP), and Val (PVP), and Ac-(Pro)(3)-Ala-Ala-(Pro)(3)-Gly Tyr-NH(2) (PAAP), by using a theoretical approach that includes a solvent effect as well as cis <--> trans isomerization of the peptide groups and puckering conformations of the pyrrolidine ring of the proline residues. Since (13)C chemical shifts have proven to be useful for identifying secondary-structure preferences in proteins and peptides, and because values of the dihedral angles (phi,psi) are the main determinants of their magnitudes, we have, therefore, computed the Boltzmann-averaged (13)C chemical shifts for the guest residues in the PXP peptide (X = Pro, Ala, Gln, Gly, and Val) with a combination of approaches, involving molecular mechanics, statistical mechanics, and quantum mechanics. In addition, an improved procedure was used to carry out the conformational searches and to compute the solvent polarization effects faster and more accurately than in previous work. The current theoretical work and additional experimental evidence show that, in short proline-rich peptides, alanine decreases the polyproline II helix content. In particular, the theoretical evidence accumulated in this work calls into question the proposal that alanine has a strong preference to adopt conformations in the polyproline II region of the Ramachandran map. PMID- 14747312 TI - Analysis of functional motions in Brownian molecular machines with an efficient block normal mode approach: myosin-II and Ca2+ -ATPase. AB - The structural flexibilities of two molecular machines, myosin and Ca(2+)-ATPase, have been analyzed with normal mode analysis and discussed in the context of their energy conversion functions. The normal mode analysis with physical intermolecular interactions was made possible by an improved implementation of the block normal mode (BNM) approach. The BNM results clearly illustrated that the large-scale conformational transitions implicated in the functional cycles of the two motor systems can be largely captured with a small number of low frequency normal modes. Therefore, the results support the idea that structural flexibility is an essential part of the construction principle of molecular motors through evolution. Such a feature is expected to be more prevalent in motor proteins than in simpler systems (e.g., signal transduction proteins) because in the former, large-scale conformational transitions often have to occur before the chemical events (e.g., ATP hydrolysis in myosin and ATP binding/phosphorylation in Ca(2+)-ATPase). This highlights the importance of Brownian motions associated with the protein domains that are involved in the functional transitions; in this sense, Brownian molecular machines is an appropriate description of molecular motors, although the normal mode results do not address the origin of the ratchet effect. The results also suggest that it might be more appropriate to describe functional transitions in some molecular motors as intrinsic elastic motions modulating local structural changes in the active site, which in turn gets stabilized by the subsequent chemical events, in contrast with the conventional idea of local changes somehow getting amplified into larger-scale motions. In the case of myosin, for example, we favor the idea that Brownian motions associated with the flexible converter propagates to the Switch I/II region, where the salt-bridge formation gets stabilized by ATP hydrolysis, in contrast with the textbook notion that ATP hydrolysis drives the converter motion. Another useful aspect of the BNM results is that selected low frequency normal modes have been identified to form a set of collective coordinates that can be used to characterize the progress of a significant fraction of large-scale conformational transitions. Therefore, the present normal mode analysis has provided a stepping-stone toward more elaborate microscopic simulations for addressing critical issues in free energy conversions in molecular machines, such as the coupling and the causal relationship between collective motions and essential local changes at the catalytic active site where ATP hydrolysis occurs. PMID- 14747313 TI - Dynamics of pinned membranes with application to protein diffusion on the surface of red blood cells. AB - We present a theoretical treatment and simulation algorithm for the dynamics of Helfrich elastic membrane surfaces in the presence of general harmonic perturbations and hydrodynamic coupling to the surrounding solvent. In the limit of localized and strong interactions, this harmonic model can be used to pin the membrane to intracellular/intercellular structures. We consider the case of pinning to the cytoskeleton and use such a model to estimate the macroscopic diffusion constant for band 3 protein on the surface of human erythrocytes. Comparison to experimental results suggests that thermal undulations of the membrane surface should play a significant role in protein mobility on the red blood cell. PMID- 14747314 TI - Opening hemichannels in nonjunctional membrane stimulates gap junction formation. AB - We studied gap junction formation in pairs of Xenopus laevis oocytes expressing connexins that form functional hemichannels and found no correlation between junctional conductance (G(j)) and whole-cell hemichannel conductances (G(hemi)) within the first few hours of pairing. However, opening hemichannels to a threshold current stimulated a rapid G(j) increase. Moreover, cx46 hemichannel current stimulated cx40 G(j) even though cx40 and cx46 do not form heteromeric or heterotypic gap junctions. Initial growth rate and final steady-state level of stimulated G(j) were proportional to the product of hemichannel conductances. External calcium affected the growth rate of stimulated G(j) but not the final steady-state value. Time constants of formation were short in low [Ca(2+)](out) (3 min in 200 micro M Ca(2+)) and long in high [Ca(2+)](out) (15 min in 1 mM Ca(2+)), but in oocyte pairs pretreated with lectins to reduce steric hindrance imposed by large membrane glycoproteins the time constant was short and Ca(2+) independent. We suggest that hemichannel activity stimulates G(j) by collapsing the extracellular volume between membranes to allow the end-to-end binding between hemichannels. These studies suggest the possibility that functional hemichannels could trigger or enhance junctional formation in vivo in response to appropriate stimuli. PMID- 14747315 TI - Mechanism of calmodulin inhibition of cardiac sarcoplasmic reticulum Ca2+ release channel (ryanodine receptor). AB - The functional effects of calmodulin (CaM) on single cardiac sarcoplasmic reticulum Ca(2+) release channels (ryanodine receptors) (RyR2s) were determined in the presence of two endogenous channel effectors, MgATP and reduced glutathione, using the planar lipid bilayer method. Single-channel activities, number of events, and open and close times were determined at varying cytosolic Ca(2+) concentrations. CaM reduced channel open probability at <10 micro M Ca(2+) by decreasing channel events and mean open times and increasing mean close times. At >10 micro M Ca(2+), CaM was less effective in inhibiting RyR2. CaM decreased mean open times but increased channel events, without significantly affecting mean close times. A series of voltage pulses was applied to the bilayer from +50 to -50 mV and from -50 mV to +50 mV to rapidly increase and decrease open channel mediated sarcoplasmic reticulum lumenal to cytosolic Ca(2+) fluxes. CaM decreased the duration of the open events after the voltage switch from -50 mV to +50 mV. In parallel experiments, a Ca(2+)-insensitive calmodulin mutant was without effect on RyR2 activity. The results are discussed in terms of a possible role of CaM in the termination of cardiac sarcoplasmic reticulum Ca(2+) release. PMID- 14747316 TI - Ca2+ modulation of Ca2+ release-activated Ca2+ channels is responsible for the inactivation of its monovalent cation current. AB - The Ca(2+) release-activated Ca(2+) (CRAC) channel is the most well documented of the store-operated ion channels that are widely expressed and are involved in many important biological processes. However, the regulation of the CRAC channel by intracellular or extracellular messengers as well as its molecular identity is largely unknown. Specifically, in the absence of extracellular divalent cations it becomes permeable to monovalent cations with a larger conductance, however this monovalent cation current inactivates rapidly by an unknown mechanism. Here we found that Ca(2+) dissociation from a site on the extracellular side of the CRAC channel is responsible for the inactivation of its Na(+) current, and Ca(2+) occupancy of this site otherwise potentiates its Ca(2+) as well as Na(+) currents. This Ca(2+)-dependent potentiation is required for the normal functioning of CRAC channels. PMID- 14747317 TI - Time-resolved charge movements in the sarcoplasmatic reticulum Ca-ATPase. AB - The time-resolved kinetics of the Ca(2+)-translocating partial reaction of the sarcoplasmatic reticulum Ca-ATPase was investigated by ATP-concentration jump experiments. ATP was released by an ultraviolet light flash from its inactive precursor and charge movements in the membrane domain of the ion pumps were detected by the fluorescent styryl dye 2BITC. Two oppositely directed cation movements were found, which were assigned to Ca(2+) release and H(+) binding. The faster process with a typical time constant of 30 ms reports the rate-limiting process before Ca(2+) release, probably the conformation transition E(1) --> E(2). The following, slow uptake of positive charge had a pH-dependent time constant, which was 1 s at low pH and approximately 3 s at pH > 8. This process is assigned to an electrically silent conformational relaxation of the state P E(2) preceding H(+) binding. This interpretation is in agreement with the observation that the fast process was independent of the substrate concentrations (i.e., when [Ca(2+)] > 200 nM, and [ATP] > 20 micro M). The slow process was independent of the Ca(2+) concentration. The activation energy of the resolved processes was between 80 kJ/mol and 90 kJ/mol, which is comparable to the activation energy of the enzymatic activity (92 kJ/mol) and these high values point to conformational changes underlying rate-limiting steps of the pump cycle. PMID- 14747318 TI - Anion pathway and potential energy profiles along curvilinear bacterial ClC Cl- pores: electrostatic effects of charged residues. AB - X-ray structures permit theoretical study of Cl(-) permeation along bacterial ClC Cl(-) pores. We determined the lowest energy curvilinear pathway, identified anion-coordinating amino acids, and calculated the electrostatic potential energy profiles. We find that all four bacterial ClC Cl(-) crystal structures correspond to closed states. E148 and S107 side chains form steric barriers on both sides of the crystal binding site in the StClC wild-type and EcClC wild-type crystals; both the EcClC(E148A) and EcClC(E148Q) mutants are blocked at the S107 site. We studied the effect that mutating the charge of some strongly conserved pore lining amino acids has on the electrostatic potential energy profiles. When E148 is neutralized, it creates an electrostatic trap, binding the ion near midmembrane. This suggests a possible electrostatic mechanism for controlling anion flow: neutralize E148, displace the side chain of E148 from the pore pathway to relieve the steric barrier, then trap the anion at midmembrane, and finally either deprotonate E148 and block the pore (pore closure) or bring a second Cl(-) into the pore to promote anion flow (pore conductance). Side-chain displacement may arise by competition for the binding site between the oxygens of E148 and the anion moving down the electrostatic energy gradient. We also find that the charge state of E111 and E113 may electrostatically control anion conductance and occupancy of the binding site within the cytoplasmic pore. PMID- 14747319 TI - Mechanism of anionic conduction across ClC. AB - ClC chloride channels are voltage-gated transmembrane proteins that have been associated with a wide range of regulatory roles in vertebrates. To accomplish their function, they allow small inorganic anions to efficiently pass through, while blocking the passage of all other particles. Understanding the conduction mechanism of ClC has been the subject of many experimental investigations, but until now, the detailed dynamic mechanism was not known despite the availability of crystallographic structures. We investigate Cl(-) conduction by means of an all-atom molecular dynamics simulation of the ClC channel in a membrane environment. Based on our simulation results, we propose a king-of-the-hill mechanism for permeation, in which a lone ion bound to the center of the ClC pore is pushed out by a second ion that enters the pore and takes its place. Although the energy required to extract the single central ion from the pore is enormous, by resorting to this two-ion process, the largest free energy barrier for conduction is reduced to 4 kcal/mol. At the narrowest part of the pore, residues Tyr-445 and Ser-107 stabilize the central ion. There, the bound ion blocks the pore, disrupting the formation of a continuous water file that could leak protons, possibly preventing the passage of uncharged solutes. PMID- 14747320 TI - Conduction mechanisms of chloride ions in ClC-type channels. AB - The conduction properties of ClC-0 and ClC-1 chloride channels are examined using electrostatic calculations and three-dimensional Brownian dynamics simulations. We create an open-state configuration of the prokaryotic ClC Cl(-) channel using its known crystallographic structure as a basis. Two residues that are occluding the channel are slowly pushed outward with molecular dynamics to create a continuous ion-conducting path with the minimum radius of 2.5 A. Then, retaining the same pore shape, the prokaryotic ClC channel is converted to either ClC-0 or ClC-1 by replacing all the nonconserved dipole-containing and charged amino acid residues. Employing open-state ClC-0 and ClC-1 channel models, current-voltage curves consistent with experimental measurements are obtained. We find that conduction in these pores involves three ions. We locate the binding sites, as well as pinpointing the rate-limiting steps in conduction, and make testable predictions about how the single channel current across ClC-0 and ClC-1 will vary as the ionic concentrations are increased. Finally, we demonstrate that a ClC-0 homology model created from an alternative sequence alignment fails to replicate any of the experimental observations. PMID- 14747321 TI - Use of cyclodextrin for AFM monitoring of model raft formation. AB - The lipid rafts membrane microdomains, enriched in sphingolipids and cholesterol, are implicated in numerous functions of biological membranes. Using atomic force microscopy, we have examined the effects of cholesterol-loaded methyl-beta cyclodextrin (MbetaCD-Chl) addition to liquid disordered (l(d))-gel phase separated dioleoylphosphatidylcholine (DOPC)/sphingomyelin (SM) and 1-palmitoyl-2 oleoyl phosphatidylcholine (POPC)/SM supported bilayers. We observed that incubation with MbetaCD-Chl led to the disappearance of domains with the formation of a homogeneously flat bilayer, most likely in the liquid-ordered (l(o)) state. However, intermediate stages differed with the passage through the coexistence of l(o)-l(d) phases for DOPC/SM samples and of l(o)-gel phases for POPC/SM bilayers. Thus, gel phase SM domains surrounded by a l(o) matrix rich in cholesterol and POPC could be observed just before reaching the uniform l(o) state. This suggests that raft formation in biological membranes could occur not only via liquid-liquid but also via gel-liquid immiscibility. The data also demonstrate that MbetaCD-Chl as well as the unloaded cyclodextrin MbetaCD make holes and preferentially extract SM in supported bilayers. This strongly suggests that interpretation of MbetaCD and MbetaCD-Chl effects on cell membranes only in terms of cholesterol movements have to be treated with caution. PMID- 14747322 TI - Correlation of AFM and SFA measurements concerning the stability of supported lipid bilayers. AB - Phospholipid bilayers were studied by means of atomic force microscopy (AFM) and a surface force apparatus (SFA). The stability of the supported bilayers was described by the amount of irregularities in the topography of the membrane by means of AFM and by the occurrence of hemifusion in the SFA, which is an indicator of defective bilayers. The bilayers, composed of lipids having the same headgroup but different chain lengths in the two leaflets, were prepared by Langmuir-Blodgett deposition and transferred at different surface pressures. The topography of the supported bilayers in aqueous solution, as imaged by AFM, revealed an increasing number of defects in the supported lipid membranes with decreased deposition pressure of the outer lipid layer. These defects, which appeared in the form of monolayer and bilayer (self-assembled) thick holes within the membrane, were energetically favorable over an evenly depleted bilayer. We found that the quantity of these defects (holes of 100:1 lead to a constant heat release and were used to directly calculate the enthalpy of the process, DeltaH, and indirectly, DeltaG and DeltaS. As the lipid/drug ratio decreased, the constancy of reaction enthalpy was tested in the fitting process. Under low lipid/drug ratio conditions simple partition was no longer valid and the interaction phenomenon was interpreted in terms of binding isotherms. A mathematical expression was deduced for quantification of the binding constants and the number of lipid molecules associated with one drug molecule. The broad range of concentrations used stressed the biphasic nature of the interaction under study. As the lipid/drug ratio was varied, the results showed that the interaction of both drugs does not present a unique behavior in all studied regimes: the extent of the interaction, as well as the binding stoichiometry, is affected by the lipid/drug ratio. The change in these parameters reflects the biphasic behavior of the interaction-possibly the consequence of a modification of the membrane's physical properties as it becomes saturated with the drug. PMID- 14747331 TI - Impedance analysis and single-channel recordings on nano-black lipid membranes based on porous alumina. AB - Ordered porous alumina substrates with pore diameters of 55 and 280 nm, respectively, were produced and utilized as a support to prepare membranes suspending the pores of the material. Highly ordered porous alumina was prepared by an anodization process followed by dissolution of the remaining aluminum and alumina at the backside of the pores. The dissolution process of Al(2)O(3) at the backside of the pores was monitored by electrical impedance spectroscopy ensuring the desired sieve-like structure of the porous alumina. One side of the porous material with an area of 7 mm(2) was coated with a thin gold layer followed by chemisorption of 1,2-dipalmitoyl-sn-glycero-3-phosphothioethanol. The hydrophobic monolayer on top of the upper surface was a prerequisite for the formation of suspending membranes, termed nano-black lipid membranes (nano-BLMs). The formation process, and long-term and mechanical stability of the nano-BLMs were followed by electrical impedance spectroscopy indicating the formation of lipid bilayers with typical specific membrane capacitances of (0.65 +/- 0.2) micro F/cm(2) and membrane resistances of up to 1.6 x 10(8) Omega cm(2). These high membrane resistances allowed for single-channel recordings. Gramicidin as well as alamethicin was successfully inserted into the nano-BLMs exhibiting characteristic conductance states. PMID- 14747332 TI - Calcium buffering and excitation-contraction coupling in developing avian myocardium. AB - This report provides a detailed analysis of developmental changes in cytoplasmic free calcium (Ca(2+)) buffering and excitation-contraction coupling in embryonic chick ventricular myocytes. The peak magnitude of field-stimulated Ca(2+) transients declined by 41% between embryonic day (ED) 5 and 15, with most of the decline occurring between ED5 and 11. This was due primarily to a decrease in Ca(2+) currents. Sarcoplasmic reticulum (SR) Ca(2+) content increased 14-fold from ED5 to 15. Ca(2+) transients in voltage-clamped myocytes after blockade of SR function permitted computation of the fast Ca buffer power of the cytosol as expressed as generalized values of B(max) and K(D). B(max) rose with development whereas K(D) did not change significantly. The computed SR Ca(2+) contribution to the Ca(2+) transient and gain factor for Ca(2+)-induced Ca(2+) release increased markedly between ED5 and 11 and slightly thereafter. These results paralleled the maturation of SR and peripheral couplings reported by others and demonstrated a strong relationship between structure and function in development of excitation contraction coupling. Modeling of buffer power from estimates of the major cytosolic Ca binding moieties yielded a B(max) and K(D) in reasonable agreement with experiment. From ED5 to 15, troponin C was the major Ca(2+) binding moiety, followed by SR and calmodulin. PMID- 14747333 TI - Activation kinetics of skinned cardiac muscle by laser photolysis of nitrophenyl EGTA. AB - The kinetics of Ca(2+)-induced contractions of chemically skinned guinea pig trabeculae was studied using laser photolysis of NP-EGTA. The amount of free Ca(2+) released was altered by varying the output from a frequency-doubled ruby laser focused on the trabeculae, while maintaining constant total [NP-EGTA] and [Ca(2+)]. The time courses of the rise in stiffness and tension were biexponential at 23 degrees C, pH 7.1, and 200 mM ionic strength. At full activation (pCa < 5.0), the rates of the rapid phase of the stiffness and tension rise were 56 +/- 7 s(-1) (n = 7) and 48 +/- 6 s(-1) (n = 11) while the amplitudes were 21 +/- 2 and 23 +/- 3%, respectively. These rates had similar dependencies on final [Ca(2+)] achieved by photolysis: 43 and 50 s(-1) per pCa unit, respectively, over a range of [Ca(2+)] producing from 15% to 90% of maximal isometric tension. At all [Ca(2+)], the rise in stiffness initially was faster than that of tension. The maximal rates for the slower components of the rise in stiffness and tension were 4.1 +/- 0.8 and 6.2 +/- 1.0 s(-1). The rate of this slower phase exhibited significantly less Ca(2+) sensitivity, 1 and 4 s(-1) per pCa unit for stiffness and tension, respectively. These data, along with previous studies indicating that the force-generating step in the cross-bridge cycle of cardiac muscle is marginally sensitive to [Ca(2+)], suggest a mechanism of regulation in which Ca(2+) controls the attachment step in the cross-bridge cycle via a rapid equilibrium with the thin filament activation state. Myosin kinetics sets the time course for the rise in stiffness and force generation with the biexponential nature of the mechanical responses to steps in [Ca(2+)] arising from a shift to slower cross-bridge kinetics as the number of strongly bound cross-bridges increases. PMID- 14747334 TI - Disruption of Caenorhabditis elegans muscle structure and function caused by mutation of troponin I. AB - Caenorhabditis elegans strains mutant for the unc-27 gene show abnormal locomotion and muscle structure. Experiments revealed that unc-27 is one of four C. elegans troponin I genes and that three mutant alleles truncate the protein: recessive and presumed null allele e155 terminates after nine codons; semidominant su142sd eliminates the inhibitory and C-terminal regions; and semidominant su195sd abbreviates the extreme C-terminus. Assays of in vivo muscular performance at high and low loads indicated that su142sd is most deleterious, with e155 least and su195sd intermediate. Microscopy revealed in mutant muscle a prevalent disorder of dense body positioning and a less well defined sarcomeric structure, with small islands of thin filaments interspersed within the overlap region of A bands and even within the H zone. The mutants' rigid paralysis and sarcomeric disarray are consistent with unregulated contraction of the sarcomeres, in which small portions of each myofibril shorten irregularly and independently of one another, thereby distorting the disposition of filaments. The exacerbated deficits of su142sd worms are compatible with involvement in vivo of the N-terminal portion of troponin I in enhancing force production, and the severe impairment associated with su195sd highlights importance of the extreme C-terminus in the protein's inhibitory function. PMID- 14747335 TI - Response of equatorial x-ray reflections and stiffness to altered sarcomere length and myofilament lattice spacing in relaxed skinned cardiac muscle. AB - Low angle x-ray diffraction measurements of myofilament lattice spacing (D(1,0)) and equatorial reflection intensity ratio (I(1,1)/I(1,0)) were made in relaxed skinned cardiac trabeculae from rats. We tested the hypothesis that the degree of weak cross-bridge (Xbr) binding, which has been shown to be obligatory for force generation in skeletal muscle, is modulated by changes in lattice spacing in skinned cardiac muscle. Altered weak Xbr binding was detected both by changes in I(1,1)/I(1,0) and by measurements of chord stiffness (chord K). Both measurements showed that, similar to skeletal muscle, the probability of weak Xbr binding at 170-mM ionic strength was significantly enhanced by lowering temperature to 5 degrees C. The effects of lattice spacing on weak Xbr binding were therefore determined under these conditions. Changes in D(1,0), I(1,1)/I(1,0), and chord K by osmotic compression with dextran T500 were determined at sarcomere lengths (SL) of 2.0 and 2.35 micro m. At each SL increasing [dextran] caused D(1,0) to decrease and both I(1,1)/I(1,0) and chord K to increase, indicating increased weak Xbr binding. The results suggest that in intact cardiac muscle increasing SL and decreasing lattice spacing could lead to increased force by increasing the probability of initial weak Xbr binding. PMID- 14747336 TI - Binding of meso-tetrakis(N-methylpyridinium-4-yl)porphyrin to AT oligomers: effect of chain length and the location of the porphyrin stacking. AB - Circular dichroism (CD) spectra of meso-tetrakis(N-methylpyridinium-4 yl)porphyrin (TMPyP) that are associated with various duplex and triplex AT oligomers were investigated in this study. A strong positive CD was apparent for both the TMPyP complexed with duplex d[(A-T)(12)](2), d(A)(12).d(T)(12) and triplex d(A)(12).d[(T)(12)](2) at a low mixing ratio. As the mixing ratio increased, bisignate excitonic CD was produced for TMPyP complexed with duplexes, whereas the positive CD signal remained the same for the TMPyP d(A)(12).d[(T)(12)](2) complex. This difference in the CD spectrum in the presence of duplex and triplex oligomers indicates that the moderate stacking of TMPyP occurs at the major groove of the duplex and the monomeric binding occurs in (or near) the minor groove. When TMPyP forms a complex with duplex d[(A T)(6)](2) only excitonic CD was observed, even at a very low mixing ratio. Therefore, at least seven or more basepairs are required for TMPyP to exhibit a monomeric CD spectrum. After close analysis of the CD spectrum, the TMPyP poly[d(A-T)(2)] complex could be explained by a combination of the CD spectrum of the monomeric, moderately stacked, and extensively stacked TMPyP. PMID- 14747337 TI - Frequency and voltage dependence of the dielectrophoretic trapping of short lengths of DNA and dCTP in a nanopipette. AB - The study of the properties of DNA under high electric fields is of both fundamental and practical interest. We have exploited the high electric fields produced locally in the tip of a nanopipette to probe the motion of double- and single-stranded 40-mer DNA, a 1-kb single-stranded DNA, and a single-nucleotide triphosphate (dCTP) just inside and outside the pipette tip at different frequencies and amplitudes of applied voltages. We used dual laser excitation and dual color detection to simultaneously follow two fluorophore-labeled DNA sequences with millisecond time resolution, significantly faster than studies to date. A strong trapping effect was observed during the negative half cycle for all DNA samples and also the dCTP. This effect was maximum below 1 Hz and decreased with higher frequency. We assign this trapping to strong dielectrophoresis due to the high electric field and electric field gradient in the pipette tip. Dielectrophoresis in electrodeless tapered nanostructures has potential applications for controlled mixing and manipulation of short lengths of DNA and other biomolecules, opening new possibilities in miniaturized biological analysis. PMID- 14747338 TI - Characterization of a novel DNA minor-groove complex. AB - Many dicationic amidine compounds bind in the DNA minor groove and have excellent biological activity against a range of infectious diseases. Para-substituted aromatic diamidines such as furamidine, which is currently being tested against trypanosomiasis in humans, and berenil, which is used in animals, are typical examples of this class. Recently, a meta-substituted diamidine, CGP 40215A, has been found to have excellent antitrypanosomal activity. The compound has a linear, conjugated linking group that can be protonated under physiological conditions when the compound interacts with DNA. Structural and molecular dynamics analysis of the DNA complex indicated an unusual AT-specific complex that involved water-mediated H-bonds between one amidine of the compound and DNA bases at the floor of the minor groove. To investigate this unique system in more detail DNase I footprinting, surface plasmon resonance biosensor techniques, linear dichroism, circular dichroism, ultraviolet-visible spectroscopy, and additional molecular dynamics simulations have been conducted. Spectrophotometric titrations of CGP 40215A binding to poly(dAT)(2) have characteristics of DNA binding-induced spectral changes as well as effects due to binding-induced protonation of the compound linker. Both footprinting and surface plasmon resonance results show that this compound has a high affinity for AT-rich sequences of DNA but very weak binding to GC sequences. The dissociation kinetics of the CGP 40215A-DNA complex are much slower than with similar diamidines such as berenil. The linear dichroism results support a minor-groove complex for the compound in AT DNA sequences. Molecular dynamics studies complement the structural analysis and provide a clear picture of the importance of water in mediating the dynamic interactions between the ligand and the DNA bases in the minor groove. PMID- 14747339 TI - Oxidation of the Mn cluster induces structural changes of NO3- functionally bound to the Cl- site in the oxygen-evolving complex of photosystem II. AB - Cl(-) is an indispensable cofactor for photosynthetic O(2) evolution and is functionally replaced by NO(3)(-). Structural changes of an isotopically labeled NO(3)(-) ion, induced by the oxidation of the Mn cluster (S(1)-to-S(2)), were detected by FTIR spectroscopy. NO(3)(-)-substituted photosystem II core particles showed (14)N(16)O(3)(-)/(15)N(16)O(3)(-) and (14)N(16)O(3)(-)/(14)N(18)O(3)(-) isotopic bands in the S(2)/S(1) spectra with markedly high signal/noise ratio. These bands appeared only in the region from 1415 to 1284 cm(-1), indicating that the bands do not arise from a metal-bound NO(3)(-) but from an ionic NO(3)(-). The intensity of the bands exhibited a quantitatively proportional relationship with the O(2) activity. These results demonstrate that the NO(3)(-) functionally bound to the Cl(-) site couples to the Mn cluster structurally, but is not associated with the cluster as a direct ligand. Comparison of the bands for two isotopes ((15)N and (18)O) and their simulations enable us to assign each band to the S(1) and S(2) states. The results indicate that the NO(3)(-) ion bound to the Cl(-) site is highly asymmetric in S(1) but rather symmetric in S(2). Since NO(3)(-) functionally replaces Cl(-), most of the conclusions drawn from this study will be also applicable to Cl(-). PMID- 14747340 TI - Recording of blue light-induced energy and volume changes within the wild-type and mutated phot-LOV1 domain from Chlamydomonas reinhardtii. AB - The time-resolved thermodynamics of the flavin mononucleotide (FMN)-binding LOV1 domain of Chlamydomonas reinhardtii phot (phototropin homolog) was studied by means of laser-induced optoacoustic spectroscopy. In the wild-type protein the early red-shifted intermediate LOV(715) exhibits a small volume contraction, DeltaV(715) = -1.50 ml/mol, with respect to the parent state. LOV(715) decays within few micro s into the covalent FMN-Cys-57 adduct LOV(390), that shows a larger contraction, DeltaV(390) = -8.8 ml/mol, suggesting a loss of entropy and conformational flexibility. The high energy content of LOV(390), E(390) = 180 kJ/mol, ensures the driving force for the completion of the photocycle and points to a strained photoreceptor conformation. In the LOV-C57S mutated protein the photoadduct is not formed and DeltaV(390) is undetected. Large effects on the measured DeltaVs are observed in the photochemically competent R58K and R58K/D31Q mutated proteins, with DeltaV(390) = -2.0 and -1.9 ml/mol, respectively, and DeltaV(715) approximately 0. The D31Q and D31N substitutions exhibit smaller but well-detectable effects. These results show that the photo-induced volume changes involve the protein region comprising Arg-58, which tightly interacts with the FMN phosphate group. PMID- 14747341 TI - FTIR difference spectroscopy in combination with isotope labeling for identification of the carbonyl modes of P700 and P700+ in photosystem I. AB - Room temperature, light induced (P700(+)-P700) Fourier transform infrared (FTIR) difference spectra have been obtained using photosystem I (PS I) particles from Synechocystis sp. PCC 6803 that are unlabeled, uniformly (2)H labeled, and uniformly (15)N labeled. Spectra were also obtained for PS I particles that had been extensively washed and incubated in D(2)O. Previously, we have found that extensive washing and incubation of PS I samples in D(2)O does not alter the (P700(+)-P700) FTIR difference spectrum, even with approximately 50% proton exchange. This indicates that the P700 binding site is inaccessible to solvent water. Upon uniform (2)H labeling of PS I, however, the (P700(+)-P700) FTIR difference spectra are considerably altered. From spectra obtained using PS I particles grown in D(2)O and H(2)O, a ((1)H-(2)H) isotope edited double difference spectrum was constructed, and it is shown that all difference bands associated with ester/keto carbonyl modes of the chlorophylls of P700 and P700(+) downshift 4-5/1-3 cm(-1) upon (2)H labeling, respectively. It is also shown that the ester and keto carbonyl modes of the chlorophylls of P700 need not be heterogeneously distributed in frequency. Finally, we find no evidence for the presence of a cysteine mode in our difference spectra. The spectrum obtained using (2)H labeled PS I particles indicates that a negative difference band at 1698 cm(-1) is associated with at least two species. The observed (15)N and (2)H induced band shifts strongly support the idea that the two species are the 13(1) keto carbonyl modes of both chlorophylls of P700. We also show that a negative difference band at approximately 1639 cm(-1) is somewhat modified in intensity, but unaltered in frequency, upon (2)H labeling. This indicates that this band is not associated with a strongly hydrogen bonded keto carbonyl mode of one of the chlorophylls of P700. PMID- 14747342 TI - End versus side branching by Arp2/3 complex. AB - We investigate the issue of end versus side branching of actin filaments by Arp2/3 complex, using a combination of analytic theory, polymerization assays, and quantitative modeling. The analytic theory shows that the effect of capping protein on the initial stages of actin polymerization in the presence of Arp2/3 complex depends strongly on whether new Arp2/3 complex-induced branches grow from the sides or ends of existing filaments. Motivated by these results, we measure and quantitatively model the kinetics of actin polymerization in the presence of activated Arp2/3 complex, for a range of concentrations of capping protein. Our model includes the most important types of events involving actin and actin binding proteins, and can be adjusted to include end branching, side branching, or both. The side-branching model gives a better fit to the experimental data than the end-branching model. An end-plus-side model including both types of branching gives a moderate improvement in the quality of the fit. Another side branching model, based on aging of subunits' capacity for branch formation, gives a significantly better fit than the end-plus-side model. We discuss implications for actin polymerization in cells. PMID- 14747343 TI - Determination of the topological shape of integral membrane protein light harvesting complex LH2 from photosynthetic bacteria in the detergent solution by small-angle X-ray scattering. AB - The topological shape of the integral membrane protein light-harvesting complex LH2 from photosynthetic bacteria Rhodobacter spheroides 2.4.1 in detergent solution has been determined from synchrotron small-angle X-ray scattering data using direct curve-fitting by the ellipsoid, ab initio shape determination methods of simulated annealing algorithm and multipole expansion, respectively. The results indicate that the LH2 protein in aqueous solution is encapsulated by a monolayered detergent shell. The detergent-stabilized structure has the shape of an oblate plate, with a thickness of 40 A, a long axis of 110 A, and a short axis of 85 A. After correction for the detergent shell, the shape of the LH2 core is also an oblate plate with a height of 40 A, a long axis of 80 A, and a short axis of 55 A. In contrast to the cylindrical crystal structure with a height of 40 A and a diameter of 68 A, the molecular shape of the LH2 complex in detergent solution clearly deviates from the ringlike crystal structure, with an eccentricity found to be 0.59-consistent with the result of single molecular spectroscopy study of the isolated single LH2 molecules. PMID- 14747344 TI - Adenylation-dependent conformation and unfolding pathways of the NAD+-dependent DNA ligase from the thermophile Thermus scotoductus. AB - In the last few years, an increased attention has been focused on NAD(+) dependent DNA ligases. This is mostly due to their potential use as antibiotic targets, because effective inhibition of these essential enzymes would result in the death of the bacterium. However, development of an efficient drug requires that the conformational modifications involved in the catalysis of NAD(+) dependent DNA ligases are understood. From this perspective, we have investigated the conformational changes occurring in the thermophilic Thermus scotoductus NAD(+)-DNA ligase upon adenylation, as well as the effect of cofactor binding on protein resistance to thermal and chemical (guanidine hydrochloride) denaturation. Our results indicate that cofactor binding induces conformational rearrangement within the active site and promotes a compaction of the enzyme. These data support an induced "open-closure" process upon adenylation, leading to the formation of the catalytically active enzyme that is able to bind DNA. These conformational changes are likely to be associated with the protein function, preventing the formation of nonproductive complexes between deadenylated ligases and DNA. In addition, enzyme adenylation significantly increases resistance of the protein to thermal denaturation and GdmCl-induced unfolding, establishing a thermodynamic link between ligand binding and increased conformational stability. Finally, chemical unfolding of deadenylated and adenylated enzyme is accompanied by accumulation of at least two equilibrium intermediates, the molten globule and premolten globule states. Maximal populations of these intermediates are shifted toward higher GdmCl concentrations in the case of the adenylated ligase. These data provide further insights into the properties of partially folded intermediates. PMID- 14747345 TI - Interhelical angle and distance preferences in globular proteins. AB - Orientational preferences between interacting helices within globular proteins have been studied extensively over the years. A number of classical structural models such as "knobs into holes" and "ridges into grooves" were developed decades ago to explain perceived preferences in interhelical angle distributions. In contrast, relatively recent works have examined statistical biases in angular distributions which result from spherical geometric effects. Those works have concluded that the predictions of classical models are due in large part to these biases. In this article we perform an analysis on the largest set of helix-helix interactions within high-resolution structures of nonhomologous proteins studied to date. We examine the interhelical angle distribution as a function of spatial distance between helix pairs. We show that previous efforts to normalize angle distribution data did not include two important effects: 1), helices can interact with each other in three distinct ways which we refer to as "line-on-line," "endpoint-to-line," and "endpoint-to-endpoint," and each of these interactions has its own geometric effects which must be included in the proper normalization of data; and 2), all normalizations that depend on geometric parameters such as interhelical angle must occur before the data is binned to avoid artifacts of bin size from biasing the conclusions. Taking these two points into account, we find that there are very pronounced preferences for helices to interact at angles of approximately +/-160 and +/-20 degrees in the line-on-line case. This pattern persists when the closest alpha-carbons in the helices vary from 4 to 12 A. The endpoint-to-line and endpoint-to-endpoint cases also exhibit distinct preferences when the data is normalized properly. Analysis of the local structural interactions which give rise to these preferences has not been studied here and is left for future work. PMID- 14747346 TI - Dielectric behavior of lysozyme and ferricytochrome-c in water/ethylene-glycol solutions. AB - This work deals with a dielectric study at radio frequencies of the influence at room temperature of two organic molecules, known as cryo-protectants, ethylene glycol and glycerol, on conformational and dynamic properties of two model proteins, lysozyme (lys) from chicken egg-white and ferricytochrome-c (cyt-c) from horse heart. Cyt-c is a compact globular protein whereas lys is composed of two structural domains, separated by the active site cleft. Measurements were carried out at the fixed temperature of 20 degrees C varying the concentration of the cosolvent up to 90% w/w. From the analysis of the dielectric relaxation of the protein solution, the effective hydrodynamic radius and the electric dipole moment of the protein were calculated as a function of the cosolvent concentration. The data show that glycerol does not modify significantly the conformation of both proteins and cyt-c is also stable in the presence of ethylene-glycol. On the contrary ethylene-glycol strongly affects the dielectric response of lysozyme denoting a specific effect on its conformation and dynamics. The data are coherently interpreted hypothesizing that glycol molecule wedges between and separates the two domains of lys making them rotationally independent. PMID- 14747347 TI - Side-chain conformational thermodynamics of aspartic acid residue in the peptides and achatin-I in aqueous solution. AB - Sequence-position dependence of the side-chain conformational equilibrium of aspartic acid (Asp) residue is investigated for both model Asp peptides (di- to tetra-) and neuropeptide achatin-I (Gly--Phe-Ala-Asp) in aqueous solution. The trans-to-gauche conformational changes on the dihedral angle of C-C(alpha) C(beta)-C are analyzed in terms of the standard free energy DeltaG(0), enthalpy DeltaH(0), and entropy -TDeltaS(0). The thermodynamic quantities are obtained by measuring the dihedral-angle-dependent vicinal (1)H-(1)H coupling constants in nuclear magnetic resonance over a wide temperature range. When the carboxyl groups of Asp are ionized, DeltaG(0) in the aqueous phase depends by approximately 1-2 kJ mol(-1) on the sequence position, whereas the energy change in the gas phase (absence of solvent) depends by tens of kJ mol(-1). Therefore, the weak position dependence of DeltaG(0) is a result of the compensation for the intramolecular effect by the hydration (= DeltaG(0)-). The DeltaH(0) and TDeltaS(0) components, on the other hand, exhibit a notable trend at the C terminus. The C-terminal DeltaH(0) is larger than the N- and nonterminal DeltaH(0) values due to the intramolecular repulsion between alpha- and beta-. The C-terminal -TDeltaS(0) is negative and larger in magnitude than the others, and an attractive solute-solvent interaction at the C-terminus serves as a structure breaker of the water solvent. PMID- 14747348 TI - Unfolding and refolding of juvenile hormone binding protein. AB - Juvenile hormone (JH) regulates insect development. JH present in the hemolymph is bound to a specific glycoprotein, juvenile hormone binding protein (JHBP), which serves as a carrier to deploy the hormone to target tissues. In this report structural changes of JHBP from Galleria mellonella induced by guanidine hydrochloride have been investigated by a combination of size-exclusion chromatography, protein activity measurements, and spectroscopic methods. Molecules of JHBP change their conformation from a native state via two unstable intermediates to a denatured state. The first intermediate appears in a compact state, because it slightly changes its molecular size and preserves most of the JHBP secondary structure of the native state. Although the second intermediate also preserves a substantial part of the secondary structure, it undergoes a change into a noncompact state changing its Stokes radius from approximately 30 to 39 A. Refolding experiments showed that JHBP molecules recover their full protein structure, as judged from the CD spectrum, fluorescence experiments, and JH binding activity measurements. The free energy of unfolding in the absence of the denaturant, DeltaG(D-N), is calculated to be 4.1 kcal mol(-1). PMID- 14747349 TI - Effects of cavity-forming mutations on the internal dynamics of azurin. AB - The effects of two single-point cavity-forming mutations, F110S and I7S, on the internal dynamics of azurin from Pseudomonas aeruginosa were probed by the phosphorescence emission of Trp-48, deeply buried in the compact hydrophobic core of the macromolecule. Changes in flexibility of the protein matrix around the chromophore were monitored by the intrinsic phosphorescence lifetime (tau(0)) whereas more general effects on structural fluctuations were deduced from the phosphorescence acrylamide quenching rate constant (k(q)), which measures the diffusion of the solute through the protein fold. The results show a spectacular, 4-5 orders of magnitude, increase of k(q) emphasizing that large amplitude structural fluctuations permitting acrylamide migration to the protein core have been drastically enhanced in each azurin mutant. The large, 12-15 kcal/mol, decrease in the activation enthalpy associated to k(q) suggests that the rate enhancement is caused, rather than through a generalized increase of protein flexibility, by the elimination of an inner barrier to the diffusion process. According to tau(0) the chromophore environment is more fluid with I7S but strikingly more rigid with F110S, demonstrating that when internal cavities are formed local effects on the mobility at the mutation site are unpredictable. Both tau(0) and k(q) reveal a structure tightening role of bound Cd(2+) that correlates with the increase in stability from apo- to holo-azurin. While these alterations in internal dynamics of azurin do not seem to play a role on electron transfer through the central region, the enhanced migration of acrylamide emphasizes that cavities may be critical for the rapid diffusion of substrates to buried, solvent inaccessible sites of enzymes. PMID- 14747350 TI - Surface functionalized cationic lipid-DNA complexes for gene delivery: PEGylated lamellar complexes exhibit distinct DNA-DNA interaction regimes. AB - Cationic lipid-DNA (CL-DNA) complexes are abundantly used in nonviral gene therapy clinical applications. Surface functionality is the next step in developing these complexes as competent, target-specific gene carriers. Poly(ethylene glycol) (PEG) is the natural choice to serve as a protective coat or act as a tether for a specific ligand on the surface of these complexes due to its biocompatibility and ability to convey stealth-like properties. Understanding the effect of PEG on the internal structure and surface properties of CL-DNA complexes is essential in developing vectors with more complex derivatives of PEG, such as Arg-Gly-Asp (RGD)-based peptide-PEG-lipids. We report on x-ray diffraction studies to probe the internal structure of CL-DNA complexes consisting of a ternary mixture of cationic lipids, neutral lipids, and PEG lipids. The PEG-coated complexes are found to exhibit a structure consistent with the lamellar phase. In addition, three distinct DNA interchain interaction regimes were found to exist, due to a), repulsive long-range electrostatic forces; b), short-range repulsive hydration forces; and c), novel polymer-induced depletion attraction forces in two dimensions. Optical microscopy and reporter gene assays further demonstrate the incorporation of the PEG-lipids into the lamellar CL-DNA complexes under biologically relevant conditions, revealing surface modification. Both techniques show that PEG-lipids with a polymer chain of molecular weight 400 do not provide adequate shielding of the PEGylated CL-DNA complexes, whereas PEG-lipids with a polymer chain of molecular weight 2000 confer stealth-like properties. This surface functionalization is a crucial initial step in the development of competent vectors for in vivo systemic gene delivery and suggests that a second type of surface functionality can be added specifically for targeting by the incorporation of peptide-PEG-lipids. PMID- 14747351 TI - Sensitivity of second harmonic generation from styryl dyes to transmembrane potential. AB - In this article we present results from the simultaneous nonlinear (second harmonic generation and two-photon excitation fluorescence) imaging and voltage clamping of living cells. Specifically, we determine the sensitivity to transmembrane potential of second harmonic generation by ANEP-chromophore styryl dyes as a function of excitation wavelength and dye structure. We have measured second harmonic sensitivities of up to 43% per 100 mV, more than a factor of four better than the nominal voltage sensitivity of the dyes under "one-photon" fluorescence. We find a dependence of voltage sensitivity on excitation wavelength that is consistent with a two-photon resonance, and there is a significant dependence of voltage sensitivity on the structure of the nonchromophore portion of the dyes. PMID- 14747352 TI - Force spectroscopy with a small dithering of AFM tip: a method of direct and continuous measurement of the spring constant of single molecules and molecular complexes. AB - A new method of direct and continuous measurement of the spring constant of single molecule or molecular complex is elaborated. To that end the standard force spectroscopy technique with functionalized tips and samples is combined with a small dithering of the tip. The change of the dithering amplitude as a function of the pulling force is measured to extract the spring constant of the complex. The potentialities of this method are illustrated for the experiments with single bovine serum albumin-its polyclonal antibody (Ab-BSA) and fibrinogen fibrinogen complexes. PMID- 14747353 TI - Localization accuracy in single-molecule microscopy. AB - One of the most basic questions in single-molecule microscopy concerns the accuracy with which the location of a single molecule can be determined. Using the Fisher information matrix it is shown that the limit of the localization accuracy for a single molecule is given by, lambda(em)/2pi n(a) square root of gammaAt, where lambda(em), n(a), gamma, A, and t denote the emission wavelength of the single molecule, the numerical aperture of the objective, the efficiency of the optical system, the emission rate of the single molecule and the acquisition time, respectively. Using Monte Carlo simulations it is shown that estimation algorithms can come close to attaining the limit given in the expression. Explicit quantitative results are also provided to show how the limit of the localization accuracy is reduced by factors such as pixelation of the detector and noise sources in the detection system. The results demonstrate what is achievable by single-molecule microscopy and provide guidelines for experimental design. PMID- 14747354 TI - Tension sensitivity of prestin: comparison with the membrane motor in outer hair cells. AB - The membrane motor in outer hair cells undergoes conformational transitions involving charge displacement of approximately 0.8 e across the membrane and changes of approximately 4 nm(2) in its membrane area. Previous reports have established that the charge transfer in the membrane motor and that in prestin, a membrane protein in the plasma membrane of outer hair cells, are approximately equal. Here, we determine the membrane area changes based on its sensitivity to membrane tension. We found that prestin does undergo area changes and that the magnitude is approximately 1 nm(2), smaller than the value 4 nm(2) for outer hair cell motor. This result confirms that prestin is a protein that functions as a membrane motor based on piezoelectricity. The discrepancy in the magnitude could suggest a prestin-containing complex in outer hair cells. PMID- 14747355 TI - Patterning, prestress, and peeling dynamics of myocytes. AB - As typical anchorage-dependent cells myocytes must balance contractility against adequate adhesion. Skeletal myotubes grown as isolated strips from myoblasts on micropatterned glass exhibited spontaneous peeling after one end of the myotube was mechanically detached. Such results indicate the development of a prestress in the cells. To assess this prestress and study the dynamic adhesion strength of single myocytes, the shear stress of fluid aspirated into a large-bore micropipette was then used to forcibly peel myotubes. The velocity at which cells peeled from the surface, V(peel), was measured as a continuously increasing function of the imposed tension, T(peel), which ranges from approximately 0 to 50 nN/ micro m. For each cell, peeling proved highly heterogeneous, with V(peel) fluctuating between 0 micro m/s ( approximately 80% of time) and approximately 10 micro m/s. Parallel studies of smooth muscle cells expressing GFP-paxillin also exhibited a discontinuous peeling in which focal adhesions fractured above sites of strong attachment (when pressure peeled using a small-bore pipette). The peeling approaches described here lend insight into the contractile-adhesion balance and can be used to study the real-time dynamics of stressed adhesions through both physical detection and the use of GFP markers; the methods should prove useful in comparing normal versus dystrophic muscle cells. PMID- 14747356 TI - Micromechanical tests of adhesion dynamics between neutrophils and immobilized ICAM-1. AB - Strong, integrin-mediated adhesion of neutrophils to endothelium during inflammation is a dynamic process, requiring a conformational change in the integrin molecule to increase its affinity for its endothelial counterreceptors. To avoid general activation of the cell, Mg(2+) was used to induce the high affinity integrin conformation, and micromechanical methods were used to determine adhesion probability to beads coated with the endothelial ligand ICAM 1. Neutrophils in Mg(2+) bind to the beads with much greater frequency and strength than in the presence of Ca(2+). An increase in adhesion strength and frequency was observed with both increasing temperature and contact duration (from 2 s to 1 min, 21 or 37 degrees C). The dependence of adhesion probability on contact time or receptor density yielded estimates of the effective reverse rate constant, k(r), and the equilibrium association constant, K(a), for binding of neutrophils to ICAM-1 coated surfaces in Mg(2+): k(r) approximately 0.7 s(-1) and the product K(a)rho(c) approximately 2.4 x 10(-4), where rho(c) is the density of integrin on the cell surface. PMID- 14747357 TI - Charged polymers modulate retrovirus transduction via membrane charge neutralization and virus aggregation. AB - The specific mechanisms of charged polymer modulation of retrovirus transduction were analyzed by characterizing their effects on virus transport and adsorption. From a standard colloidal perspective two mechanisms, charge shielding and virus aggregation, can potentially account for the experimentally observed changes in adsorption behavior and biophysical parameters due to charged polymers. Experimental testing revealed that both mechanisms could be at work depending on the characteristics of the cationic polymer. All cationic polymers enhanced adsorption and transduction via charge shielding; however, only polymers greater than 15 kDa in size were capable of enhancing these processes via the virus aggregation mechanism, explaining the higher efficiency enhancement of the high molecular weight molecules. The role of anionic polymers was also characterized and they were found to inhibit transduction via sequestration of cationic polymers, thereby preventing charge shielding and virus aggregation. Taken together, these findings suggest the basis for a revised physical model of virus transport that incorporates electrostatic interactions through both virus-cell repulsive and attractive interactions, as well as the aggregation state of the virus. PMID- 14747358 TI - Relationship between molecular and cellular dissociation rates for VLA-4/VCAM-1 interaction in the absence of shear stress. AB - The rate of leukocyte recruitment to and detachment from the vasculature contributes to cellular tethering, rolling, firm adherence, and migration across an endothelium layer. The molecular rates depend on the type and number of bound integrin or selectin adhesion molecules, shear force acting on the bound adhesion molecules, and affinity state of integrins. Although little is known of the effect that the number of adhesion molecules has on leukocyte recruitment, it has been shown that firm adhesion for cells in suspension may be mediated by small numbers of bound adhesion molecules. We studied the disaggregation of aggregates composed of B78H1 cells transfected with human vascular cell adhesion molecule-1 (VCAM-1) and human monoblastoid U937 cells expressing Very Late Antigen-4 (VLA 4). Aggregate disaggregation rates were obtained and compared to dissociation rates for soluble rhVCAM-1 ligand and monoblastoid U937 cells. Under conditions without shear stress, it was found that average cellular disaggregation rates were a factor of 1.3 +/- 0.4 times slower than molecular dissociation rates for the 1 mM Mn(2+) and 1 mM Mn(2+) + 1 mM Ca(2+) conditions. A simple mathematical model was used to predict how much smaller the dissociation constant would be if the number of bonds holding an aggregate varied from one bond to N bonds under conditions without shear stress. The average number of adhesion bonds holding the cell aggregates together was found to be 1.5 +/- 0.7. This suggests that a few bonds were needed to form cellular aggregates and that increased aggregation was related to integrin affinity changes and not due to clustering or increased bond numbers. PMID- 14747359 TI - Temperature dependency of molecular mobility in preserved seeds. AB - Although cryogenic storage is presumed to provide nearly infinite longevity to cells, the actual timescale for changes in viability has not been addressed theoretically or empirically. Molecular mobility within preserved biological materials provides a first approximation of the rate of deteriorative reactions that ultimately affect shelf-life. Here, temperature effects on molecular mobility in partially dried seeds are calculated from heat capacities, measured using differential scanning calorimetry, and models for relaxation of glasses based on configurational entropy. Based on these analyses, glassy behavior in seeds containing 0.07 g H(2)O/g dm followed strict Vogel-Tamman-Fulcher (VTF) behavior at temperatures above and just below the glass transition temperature (Tg) at 28 degrees C. Temperature dependency of relaxation times followed Arrhenius kinetics as temperatures decreased well below Tg. The transition from VTF to Arrhenius kinetics occurred between approximately 5 and -10 degrees C. Overall, relaxation times calculated for seeds containing 0.07 g H(2)O/g dm decreased by approximately eight orders of magnitude when seeds were cooled from 60 to -60 degrees C, comparable to the magnitude of change in aging kinetics reported for seeds and pollen stored at a similar temperature range. The Kauzmann temperature (T(K)), often considered the point at which molecular mobility of glasses is practically nil, was calculated as -42 degrees C. Calculated relaxation times, temperature coefficients lower than expected from VTF kinetics, and T(K) that is 70 degrees C below Tg suggest there is molecular mobility, albeit limited, at cryogenic temperatures. PMID- 14747361 TI - A randomized, controlled pilot trial of a school intervention for children with sickle cell anemia. AB - OBJECTIVE: To investigate the feasibility and efficacy of a randomized clinical pilot trial comparing routine services (RS) with a school intervention program (SIP) for children with sickle cell anemia (SCA). METHOD: Twenty-four children (ages 8-12 years) with SCA were randomized to RS or SIP. General disease knowledge, consumer satisfaction, self-concept, and school absences were evaluated. RESULTS: Compared with children receiving RS, children and teachers receiving SIP evidenced more accurate information about their disease, and children with SCA had significantly fewer absences. Teachers receiving SIP reported higher consumer satisfaction. CONCLUSIONS: A modest educational curriculum can increase knowledge of SCA, is associated with lower absence rates, and yields high consumer satisfaction ratings. PMID- 14747362 TI - The role of fathers in toddlers' unintentional injury risk. AB - OBJECTIVES: Previous work suggests that maternal parenting and supervision reduces risk for children's unintentional injuries, but very little research has examined the role of fathers in children's unintentional injury risk. The role of fathers in protecting children from unintentional injury was considered. METHODS: A prospective longitudinal design predicted injury risk in 181 toddlers from the ages of 6 to 36 months. Predictor variables included child gender and temperament, individual difference factors of the mother and father, and parenting factors of the mother and father. RESULTS: Hierarchical regression models suggested that fathers' report of gains to the family from their employment was the strongest predictor of risk of children's unintentional injury. Several other paternal and maternal factors were also modestly related to injury risk. CONCLUSIONS: Data indicate the need to consider the role of fathers in protecting children from unintentional injuries. PMID- 14747363 TI - Peer rejection, social behavior, and psychological adjustment in children with juvenile rheumatic disease. AB - OBJECTIVE: To examine the associations among disease status, social competence, and depressive symptoms in children with juvenile rheumatic disease (JRD) and to test the hypothesis that individual differences in children's social competence account for a significant proportion of variance in depressive symptoms after controlling for disease status variables. METHOD: Thirty-six children with JRD completed standardized instruments to assess pain, health status, and depressive symptoms. The rheumatologist completed a disease severity measure, and teachers provided ratings of peer rejection and social behavior. RESULTS: Pain, peer rejection, and problematic social behavior were all positively associated with depressive symptoms. Social variables remained significantly associated with depressive symptoms after controlling for level of pain. In addition, peer rejection moderated the association between pain and depressive symptoms, such that children with high levels of pain and high levels of peer rejection reported the highest frequency of depressive symptoms. CONCLUSIONS: Health care providers should assess the social functioning of children with JRD in order to identify socially vulnerable children who may be at increased risk for internalizing problems. PMID- 14747364 TI - The role of autonomy and pubertal status in understanding age differences in maternal involvement in diabetes responsibility across adolescence. AB - OBJECTIVE: To examine how autonomy and pubertal status explain age decreases in maternal involvement in type 1 diabetes management across adolescence, how they relate to metabolic control, and the reasons that guide declines in maternal involvement. METHODS: One hundred twenty-seven children ages 10-15 years with type 1 diabetes and their mothers participated. Data included maternal and child report of diabetes management, child report of autonomy level, maternal report of pubertal status, maternal reports of reasons for transfer of diabetes responsibility, and glycosylated hemoglobin (Hba(1c)) values. RESULTS: Autonomy and pubertal status partially mediated age effects on reports of maternal involvement. Mothers' reasons for transferring responsibility included responding to the child's competence, promoting competence and maturity in their child, and minimizing hassles and conflict. The transfer of diabetes responsibility from mother to child without sufficient autonomy and when pubertal status was low was related to higher Hba(1c) values. CONCLUSIONS: The importance of chronological age for changes in maternal involvement suggests the need to examine mothers' and adolescents' developmental expectations for diabetes management. The reasons for transferring responsibility from mother to child suggest many avenues for intervention. PMID- 14747365 TI - 2000 SPP Salk Award Address. Financing pediatric psychology services: buddy, can you spare a dime? PMID- 14747366 TI - First, AIDE for pediatric psychology. PMID- 14747367 TI - Financing pediatric psychology services: "Look what they've done to my song, ma" or "The sun'll come out tomorrow"? PMID- 14747368 TI - We can make our own dime or two, help children and their families, and advance science while doing so. PMID- 14747369 TI - Financing pediatric psychology: on "Buddy, can you spare a dime?". PMID- 14747370 TI - The classification of glomerulonephritis in systemic lupus erythematosus revisited. AB - The currently used classification reflects our understanding of the pathogenesis of the various forms of lupus nephritis, but clinicopathologic studies have revealed the need for improved categorization and terminology. Based on the 1982 classification published under the auspices of the World Health Organization (WHO) and subsequent clinicopathologic data, we propose that class I and II be used for purely mesangial involvement (I, mesangial immune deposits without mesangial hypercellularity; II, mesangial immune deposits with mesangial hypercellularity); class III for focal glomerulonephritis (involving <50% of total number of glomeruli) with subdivisions for active and sclerotic lesions; class IV for diffuse glomerulonephritis (involving > or = 50% of total number of glomeruli) either with segmental (class IV-S) or global (class IV-G) involvement, and also with subdivisions for active and sclerotic lesions; class V for membranous lupus nephritis; and class VI for advanced sclerosing lesions]. Combinations of membranous and proliferative glomerulonephritis (i.e., class III and V or class IV and V) should be reported individually in the diagnostic line. The diagnosis should also include entries for any concomitant vascular or tubulointerstitial lesions. One of the main advantages of the current revised classification is that it provides a clear and unequivocal description of the various lesions and classes of lupus nephritis, allowing a better standardization and lending a basis for further clinicopathologic studies. We hope that this revision, which evolved under the auspices of the International Society of Nephrology and the Renal Pathology Society, will contribute to further advancement of the WHO classification. PMID- 14747371 TI - N-acetylcysteine in the prevention of radiocontrast-induced nephropathy. AB - N-acetylcysteine is a remarkably active agent shown to be useful in a variety of clinical settings. The drug has actions relevant to radiocontrast-induced nephropathy (RCIN) that include vasodilatation, enhancement of renal medullary blood flow, and antioxidant properties. The drug's pharmacokinetics are remarkable for almost complete first pass metabolism after oral administration, resulting in no free drug reaching the circulation. After intravenous administration, extensive reaction with tissue and plasma proteins greatly limits the amount of circulating free drug. Given the difficulty achieving free drug in the systemic circulation, it is highly likely that the drug works via its metabolites. The primary mechanism may be through L-cysteine as a cellular source for glutathione production. Clinical studies of N-acetylcysteine in the prevention of RCIN have yielded highly mixed results; five were dramatically positive, and eight others had no demonstrable efficacy at all. The following will review the individual studies, attempt to reconcile the divergent results, and propose future research needs. PMID- 14747372 TI - Function and localization of urate transporter 1 in mouse kidney. AB - Mouse renal-specific transporter (RST) cDNA, the amino acid sequence of which has 74% identity with that of human urate transporter 1 (hURAT1), is potentially the mouse homologue of hURAT1, the gene responsible for hereditary renal hypouricemia. The aim of this study is to determine the location and characteristics of RST molecule in mouse kidney and investigate urate transport by RST using the Xenopus oocyte expression system. RST transported (14)C-urate in a Michaelis-Menten manner. The K(m) and the V(max) values of RST-dependent urate transport were 1213 +/- 222 micro M and 268.8 +/- 38.0 pmol/oocyte per hr, respectively (n = 3). RST-dependent urate transport was cis-inhibited significantly by 1 mM probenecid (68.7 +/- 9.4%), 50 micro M benzbromarone (67.9 +/- 6.4%), and 10 mM lactate (50.9 +/- 9.5%). However, 1 mM p-aminohippurate (PAH), 1 mM xanthine, and 1 mM oxonate did not inhibit RST-dependent urate transport. Substitution of Cl anion with gluconate in the external solution enhanced RST-dependent urate transport. Pre-injected pyrazinoic acid (PZA) or L lactate trans-stimulated RST-dependent urate transport. Using immunohistochemistry for mouse kidney, the brush border or intracellular membrane of proximal tubules was stained by an affinity-purified antibody that recognized mouse URAT1 (mURAT1) expressed on Xenopus oocyte. Using Western blotting, anti mURAT1 antibody detected 70-kD and 62-kD protein bands. The 70-kD protein was N glycosylated and was identified as a Triton X-100 insoluble brush border membrane protein. RST mRNA and protein levels were higher in male kidneys than female. RST transported urate similar to hURAT1 and, therefore, appears to be mURAT1-the mouse homologue of hURAT1. PMID- 14747373 TI - Acute and chronic regulation of thick ascending limb endothelial nitric oxide synthase by statins. AB - Statins, 3-hydroxy-3-methylglutaryl CoA reductase inhibitors, acutely increase endothelial nitric oxide synthase (eNOS) activity and chronically increase eNOS expression in endothelial cells. NO decreases transport in thick ascending limbs (TAL). We hypothesized that statins inhibit TAL transport by acutely activating eNOS, thereby increasing NO production and chronically enhancing eNOS expression. Oxygen consumption (QO(2)) by TAL suspensions from Sprague-Dawley rats was used as a measure of active NaCl reabsorption. Na/K ATPase activity was assessed by measuring ATP hydrolysis in the presence and absence of ouabain. eNOS expression was measured by Western blot. A total of 50 micro M pravastatin decreased QO(2) by 18.6 +/- 3.4% (P < 0.01). In the presence of 500 micro M furosemide and 200 micro M amiloride, transport blockers, QO(2) remained the same after pravastatin was added. Na/K ATPase activity was not different from controls and TAL treated with 50 micro M pravastatin (0.33 +/- 0.07 versus 0.29 +/- 0.04 nmol P(i)/ micro g protein/min, where P(i) is inorganic phosphate). Nystatin stimulated QO(2) to 178 +/- 13.7 in pravastatin-treated TAL and 195 +/- 11.5 in furosemide-treated TAL. The inhibitory effect of pravastatin on QO(2) was blocked by L-nitroarginine methyl ester, an NOS inhibitor. In addition, pravastatin increased NO production as measured by the fluorescent dye DAF-2A. Pravastatin at a dose of 10 mg/kg per d had no effect on eNOS protein at 1 d (24.1 +/- 2.7 versus 25.5 +/- 1.1 arbitrary units [AU]) or 7 d (24.1 +/- 2.7 versus 20.9 +/- 1.3 AU). Similarly, at 1 d, 50 mg/kg per d had no effect on expression (24.1 +/- 2.7 versus 21.2 +/- 3.6 AU). At 7 d, this dose decreased eNOS protein from 24.1 +/- 2.7 to 11.8 +/- 4.4 AU. It is concluded that pravastatin acutely decreases NaCl entry into the TAL by releasing NO. Pravastatin does not chronically increase eNOS expression in TAL. PMID- 14747374 TI - Activation of the signal transducer and activator of transcription signaling pathway in renal proximal tubular cells by albumin. AB - Renal proximal tubular cells activated by reabsorption of protein are thought to play significant roles in the progression of kidney diseases. It was hypothesized that the signal transducer and activator of transcription (STAT) proteins may be activated by proteinuria in proximal tubular cells. To test this hypothesis, murine proximal tubular cells were treated with albumin (30 mg/ml medium) for various lengths of time. The results showed that albumin could activate Stat1 and Stat5 within 15 min in proximal tubular cells. The activation of STATs was mediated mostly by Jak2 and required no protein synthesis. In addition, activation of Stat1 occurred even after neutralization of IFN-gamma. The activation of STATs was inhibited by N-acetyl-L-cysteine, a precursor of glutathione and a reactive oxygen species (ROS) scavenger, and fluorescence activated cell sorter analysis showed upregulation of intracellular ROS after albumin overloading, suggesting that albumin per se could generate ROS in proximal tubular cells. The activation of STATs occurred by way of the ROS generating system, and especially through the membrane-bound NADPH oxidase system. Reduced activities of glutathione peroxidase and catalase could also be responsible for the accumulation of intracellular ROS. Hence, not only the ROS generating system, but also the ROS scavenging system may contribute to the induction of ROS by albumin. These findings support the hypothesis that proximal tubular cells are activated and generate ROS by reabsorption of abundant urinary proteins filtered through the glomerular capillaries, and as a consequence, various IFN-gamma-inducible proteins are synthesized through IFN-gamma independent activation of STAT signaling. PMID- 14747375 TI - Exogenous PDGF-D is a potent mesangial cell mitogen and causes a severe mesangial proliferative glomerulopathy. AB - The PDGF family consists of at least four members, PDGF-A, -B, -C, and -D. All of the PDGF isoforms bind and signal through two known receptors, PDGF receptor alpha and PDGF receptor-beta, which are constitutively expressed in the kidney and are upregulated in specific diseases. It is well established that PDGF-B plays a pivotal role in the mediation of glomerular mesangial cell proliferation. However, little is known of the roles of the recently discovered PDGF-C and -D in mediating renal injury. In this study, adenovirus constructs encoding PDGF-B, -C, and -D were injected into mice. Mice with high circulating levels of PDGF-D developed a severe mesangial proliferative glomerulopathy, characterized by enlarged glomeruli and a striking increase in glomerular cellularity. The PDGF-B overexpressing mice had a milder proliferative glomerulopathy, whereas the mice overexpressing PDGF-C and those that received adenovirus alone showed no measurable response. Mitogenicity of PDGF-D and -B for mesangial cells was confirmed in vitro. These findings emphasize the importance of engagement of PDGF receptor-beta in transducing mesangial cell proliferation and demonstrate that PDGF-D is a major mediator of mesangial cell proliferation. Finally, this approach has resulted in a unique and potentially valuable model of mesangial proliferative glomerulopathy and its resolution. PMID- 14747376 TI - Rescue of defective branching nephrogenesis in renal-coloboma syndrome by the caspase inhibitor, Z-VAD-fmk. AB - In renal-coloboma syndrome (RCS), null mutations of the PAX2 gene cause renal hypoplasia due to a congenital deficit of nephrons; affected individuals may develop renal insufficiency in childhood. During normal kidney development, PAX2, is expressed at high levels throughout the arborizing ureteric bud (UB); recent observations suggest that one of its key roles is to suppress apoptosis in this collecting duct lineage. The authors hypothesized that increased UB cell apoptosis due to PAX2 haploinsufficiency must directly influence the rate of branching morphogenesis in developing kidney and the number of nephrons that can be formed before birth, when nephrogenesis in humans comes to an end. If so, the authors reasoned that caspase inhibitors might be used to suppress unwanted UB cell apoptosis during kidney development in Pax2(1Neu) mutant mice and rescue the genetic UB branching defect. E17.5 kidneys from Pax2(1Neu) mutant mice had smaller (-25%) longitudinal cross-sectional area and 3.5-fold increase in collecting duct cell apoptosis versus wild-type littermates; mutant E13.5 kidney explants allowed to arborize for 50 h in vitro had 18% fewer terminal branches than wild-types. However, exposure to the caspase inhibitor, Z-VAD-fmk (25 micro M), significantly increased terminal branch number in mutant explants (23%). It also increased branching in wild-type explants, apparently reflecting an effect of Z-VAD-fmk on basal apoptosis induced by ex vivo culture conditions. Similarly, when pregnant mice were injected daily with Z-VAD-fmk (10 micro g/g weight from E10.5 to E17.5), apoptosis of Pax2(1Neu) fetal collecting duct cells was suppressed to 40% of untreated mutants; by E14, terminal branch number was increased to 152% that of untreated litters. These studies support the hypothesis that PAX2 normally optimizes the rate of branching morphogenesis in fetal kidney by suppressing UB apoptosis. Furthermore, it suggests that caspase inhibitors can rescue the branching defect caused by PAX2 mutations. PMID- 14747377 TI - The SOD mimetic tempol ameliorates glomerular injury and reduces mitogen activated protein kinase activity in Dahl salt-sensitive rats. AB - It was shown recently that renal injury in Dahl salt-sensitive (DS) hypertensive rats is accompanied by mitogen-activated protein kinase (MAPK) activation. The present study was conducted to elucidate the contribution of reactive oxygen species to MAPK activities and renal injury in DS rats. DS rats were maintained on high salt (H; 8.0% NaCl; n = 7) or low salt (L; 0.3% NaCl; n = 6) diets; H + a superoxide dismutase mimetic, tempol (3 mmol/L in drinking water; n = 8); or H + hydralazine (0.5 mmol/L in drinking water; n = 8) for 4 wk. Mean BP (MBP) in DS/H and DS/L rats was 185 +/- 7 and 113 +/- 3 mmHg, respectively. DS/H rats showed a higher ratio of urinary protein excretion and creatinine (U(protein)V/U(cr)V; 20.3 +/- 1.1) and a higher cortical collagen content (22 +/- 1 micro g/mg) than in DS/L rats (2.4 +/- 0.1 and 13 +/- 1 micro g/mg, respectively). The expression of p22-phox and Nox-1, essential components of NAD(P)H oxidase, in renal cortical tissue was approximately threefold higher in DS/H rats than in DS/L rats. Increased activities of renal cortical MAPK, including extracellular signal regulated kinases (ERK) 1/ERK2 and c-Jun NH(2)-terminal kinases (JNK) were also observed in DS/H rats by 7.0 +/- 0.7- and 4.3 +/- 0.2-fold, respectively. Tempol treatment significantly decreased MBP (128 +/- 3 mmHg), U(protein)V/U(cr)V (4.8 +/- 0.4), and cortical collagen content (14 +/- 1 micro g/mg) and normalized ERK1/ERK2 and JNK activities in DS/H rats. Histologically, tempol markedly ameliorated progressive sclerotic and proliferative glomerular changes in DS/H rats. Hydralazine-treated DS/H rats showed similar MBP (127 +/- 5 mmHg) to tempol treated DS/H rats. Hydralazine also decreased U(protein)V/U(cr)V (16.2 +/- 1.5) and cortical collagen content (19 +/- 1 micro g/mg) in DS/H rats. However, these values were significantly higher than those of tempol-treated rats. Furthermore, although hydralazine significantly reduced JNK activity (-56 +/- 3%), ERK1/ERK2 activities were unaffected. These data suggest that reactive oxygen species, generated by NAD(P)H oxidase, contribute to the progression of renal injury through ERK1/ERK2 activation in DS/H hypertensive rats. PMID- 14747378 TI - beta(2)-Adrenoceptor activation attenuates endotoxin-induced acute renal failure. AB - Abnormalities in the beta(2)-adrenergic control of organ function have been implicated in the pathogenesis of several disease states, such as septic shock. The objectives of the present study were to define the contribution of beta(2) adrenoceptors (beta(2)-AR) to normal renal physiology and to investigate whether overexpression of renal beta(2)-AR might be potentially beneficial in preventing progressive renal damage associated with endotoxemia. Adenoviral transgenes containing the human beta(2)-AR (Adeno-beta(2)-AR) were constructed and delivered into the rat kidney by means of intraparenchymal injections. Administration of 10(9) total viral particles of Adeno-beta(2)-AR induced an approximately threefold increase in beta(2)-AR density in the renal tissue, which 2 wk after delivery, enhanced GFR and sodium reabsorption compared with control rats. The enhanced GFR was abolished by the addition of the beta(2)-AR antagonist, ICI 118,551. Administration of lipopolysaccharide (LPS) caused a reduction in GFR, beta(2)-AR density, and cAMP together with enhanced TNF-alpha mRNA in the kidney. In rats overexpressing beta(2)-AR, the reduction in baseline GFR and elevation of TNF-alpha mRNA and leukocyte infiltration into the kidney associated with the endotoxin were blocked. These findings suggested the possibility that a renal specific overexpression of beta(2)-AR preserves basal renal function in response to a ligand-independent beta(2)-AR activation and that the delivery of Adeno beta(2)-AR gene is a potential novel therapeutic strategy for treatment of acute renal failure associated with sepsis. PMID- 14747379 TI - p38 Mitogen-activated protein kinase activation and cell localization in human glomerulonephritis: correlation with renal injury. AB - Activation of the p38 mitogen-activated protein kinase (MAPK) signal transduction pathway plays an important role in the inflammatory response. It was postulated that p38 MAPK is important in the pathogenesis of human glomerulonephritis and contributes to the development of renal injury. p38 MAPK activation was examined by immunodetection for dual phosphorylated p38 (p-p38) in normal human kidney and 77 renal biopsy specimens encompassing a wide spectrum of glomerulonephritides. In normal kidney, p-p38 immunostaining was restricted to the nuclei of a small number of podocytes, parietal epithelial cells, and tubular cells. There was a dramatic increase in the number of p-p38-positive cells in glomeruli and tubules in nonproliferative and proliferative glomerulonephritis and a substantial increase in the number of interstitial p-p38-positive cells in proliferative glomerulonephritis. Double immunostaining identified p38 activation in intrinsic renal cells (podocytes and endothelial and tubular cells), infiltrating macrophage and neutrophils, and myofibroblasts. Renal failure correlated with the number of p-p38-positive glomerular, tubular, and interstitial cells. Proteinuria correlated with the number of p-p38-positive tubular and interstitial cells and the number of p-p38-positive podocytes in nonproliferative glomerulonephritis. Furthermore, glomerular p38 activation correlated with segmental proliferative and necrotic lesions, and interstitial p38 activation correlated with the degree of interstitial inflammation. In conclusion, activation of p38 MAPK in intrinsic renal cells and infiltrating leukocytes correlated with renal dysfunction and histopathology, suggesting an important pathogenic role for p38 MAPK activation in human glomerulonephritis. PMID- 14747380 TI - Chemokine receptor CCR1 but not CCR5 mediates leukocyte recruitment and subsequent renal fibrosis after unilateral ureteral obstruction. AB - As chemokine receptor CCR1 and CCR5 expression on circulating leukocytes is thought to contribute to leukocyte recruitment during renal fibrosis, the authors examined the effects of unilateral ureteral obstruction (UUO) in mice deficient for CCR1 or CCR5. Analysis of UUO kidneys from CCR1-deficient mice revealed a reduction of interstitial macrophages and lymphocytes (35% and 55%, respectively) compared with wild-type controls. CCR1-deficient mice had reduced CCR5 mRNA levels in UUO kidneys, which correlated with a reduction of CCR5+ T cell infiltrate as determined by flow cytometry. Interstitial fibroblasts, renal TGF beta1 mRNA expression, interstitial volume, and collagen I deposits were all significantly reduced in CCR1-deficient mice. In contrast, renal leukocytes and fibrosis were unaffected in CCR5-deficient mice with UUO. However, if treated with the CCR1 antagonist BX471, CCR5-deficient mice showed a similar reduction of renal leukocytes and fibrosis as CCR1-deficient mice. To determine the underlying mechanism labeled macrophages and T cells isolated from either wild-type, CCR1 deficient, or CCR5-deficient mice were injected into wild-type mice with UUO. Three hours later, renal cell recruitment was reduced for CCR1-deficient cells or cells pretreated with BX471 compared with CCR5-deficient or wild-type cells. Thus, CCR1 but not CCR5 is required for leukocyte recruitment and fibrosis after UUO in mice. Therefore, CCR1 is a promising target for therapeutic intervention in leukocyte-mediated fibrotic tissue injury, e.g. progressive renal fibrosis. PMID- 14747381 TI - Beneficial Effects of Estrogens on Indices of Renal Damage in Uninephrectomized SHRsp Rats. AB - Renal diseases tend to be less severe among premenopausal female patients, compared with male patients. Experimental data on the effects of estrogens on renal damage are controversial, and potential underlying mechanisms have not been fully clarified. Three-month-old, female, uninephrectomized (UNX), sham-operated or ovariectomized (OVX) SHRsp rats were left untreated or received either 17beta estradiol 3-benzoate (25 micro g/d) or estriol (0.02 mg/d) daily. After 3 mo, indices of renal damage (glomerulosclerosis index and tubulointerstitial damage index) and glomerular geometric parameters were investigated. The expression of desmin, TGF-beta, endothelin-1, collagen IV, endothelial nitric oxide synthase, and estrogen receptors alpha and beta in the glomeruli and tubulointerstitium was immunohistochemically evaluated. Estradiol and estriol did not significantly affect kidney weights or BP. Estradiol and estriol caused significant reductions in albuminuria (vehicle-treated UNX/OVX animals, 25.4 +/- 8.52 mg/24 h; estradiol treated UNX/OVX animals, 15.37 +/- 6.12 mg/24 h; estriol-treated UNX/OVX animals, 6.54 +/- 2.24 mg/24 h). The glomerulosclerosis index was significantly lower in estriol- and estradiol-treated animals (estradiol-treated UNX/OVX animals, 0.69 +/- 0.16; estriol-treated UNX/OVX animals, 0.21 +/- 0.12; P < 0.05), compared with vehicle-treated animals (1.46 +/- 0.09); the tubulointerstitial damage index exhibited a similar pattern. The mean glomerular volume was significantly less in estrogen-treated animals. UNX/OVX animals demonstrated significantly greater expression of TGF-beta and endothelin-1 in immunohistochemical, in situ hybridization, and reverse transcription-PCR assays. This increase was abrogated by estriol but not estradiol. Similarly, significantly higher glomerular and tubulointerstitial expression of proliferating cell nuclear antigen and collagen IV was observed in UNX/OVX animals, and expression was decreased by estriol but not estradiol. It was concluded that, in the UNX model of spontaneous renal damage, glomerular lesions and glomerular hypertrophy were reduced by estriol but less consistently by estradiol. In parallel, loss of podocytes, evidence of podocyte injury (i.e., desmin expression), and expression of mediator systems of glomerular damage were decreased, pointing to a major renoprotective action of estriol. PMID- 14747382 TI - Successful treatment of an adynamic bone disorder with bone morphogenetic protein 7 in a renal ablation model. AB - An adynamic bone disorder (ABD) is an important complication of chronic kidney disease (CKD) of unknown etiology for which there is no adequate treatment. Reported is an animal model of ablative CKD complicated by an ABD characterized by the absence of secondary hyperparathyroidism and its successful treatment with a skeletal anabolic factor, bone morphogenetic protein-7 (BMP-7). Adult mice were subjected to electrocautery of the right kidney followed by left nephrectomy. Animals were randomized into groups fed normal chow or fed low-phosphate chow supplemented with calcitriol to maintain normophosphatemia in CKD. All groups were maintained on the regimens for 12 wk. Hyperphosphatemia, secondary hyperparathyroidism, and a mild osteodystrophy developed in the CKD/chow-fed group, as expected. When dietary phosphorus was restricted and calcitriol was administered in the CKD low-phosphate/calcitriol group (ABD), Ca, PO(4), and parathyroid hormone levels were maintained normal. A significant ABD developed in the ABD group characterized by significant depressions in osteoblast number, perimeters, bone formation rates, and mineral apposition rates when compared with the sham-operated, chow-fed group. The abnormal skeletal histomorphometry was reversed by BMP-7 therapy to normal values and significantly improved from the ABD group (P < 0.05). The sham-operated low-phosphate/calcitriol-fed control group and the CKD low-phosphate/calcitriol/BMP-7 groups had reduced phosphate levels compared with the other groups (P < 0.05). ABD produced in mice with CKD in the absence of hyperparathyroidism was successfully reversed with a bone anabolic, BMP-7, associated with a reduction in plasma phosphorus. PMID- 14747383 TI - The role of p38alpha mitogen-activated protein kinase activation in renal fibrosis. AB - The p38 mitogen-activated protein kinase (MAPK) pathway transduces external stress stimuli and is important in extracellular matrix synthesis in cell types in vitro; however, its role in renal fibrosis is not known. Explored was the role the p38 MAPK pathway in rat unilateral ureteric obstruction (UUO), a model of renal fibrosis induced by a noninflammatory surgical insult. In a time-course study, a marked increase in phosphorylation (activation) of p38 in both interstitial myofibroblasts and tubules was shown. Rats were then treated daily with a specific inhibitor of p38alpha, NPC 31169, from the time of UUO surgery until being killed 7 d later. Compared with vehicle, NPC 31169-treated rats had a significant reduction in renal fibrosis assessed by interstitial volume, collagen IV deposition, and mRNA levels. This was primarily due to a reduction in the accumulation of interstitial myofibroblasts, as shown by a reduction in the area of immunostaining for alpha-smooth muscle actin and heat shock protein 47. The increase in renal TGF-beta1 mRNA and protein levels in UUO was unaltered with NPC 31169 treatment; however, connective tissue growth factor mRNA was reduced. These results demonstrate that p38alpha MAPK plays an important role in renal fibrosis, acting downstream of TGF-beta1. Blockade of p38 MAPK reduces extracellular matrix production and may be considered a potential therapeutic option in the treatment of renal fibrosis. PMID- 14747385 TI - Chronic renal allograft dysfunction: the role of T cell-mediated tubular epithelial to mesenchymal cell transition. AB - Chronic graft dysfunction is now the leading clinical problem after renal transplantation. The principal histopathologic lesion seen in this disease is tubular loss with concurrent interstitial fibrosis. Although the severity of acute rejection often correlates with that of subsequent chronic dysfunction, a direct link between these processes has remained elusive. This study was designed to test the hypothesis that intraepithelial T cells recruited to the renal tubules during acute rejection can directly induce fibrosis by causing tubular epithelial cells to undergo transformation to produce a motile population of activated fibroblasts. A study of renal allograft tissue sections showed the presence of the S100A4 marker of epithelial to mesenchymal transition in some tubular epithelial cells; the expression of this antigen was heterogeneous both within and between individual tubular cross-sections. Significantly, S100A4 expressing epithelial cells were frequently associated with infiltrating CD8+ T cells, and many coexpressed the Ki67 marker of proliferation. A parallel study of human renal cortical epithelial cells in primary culture demonstrated that S100A4 was induced by stimulation for 72 h with TGF-beta(1) and by direct contact with membrane-associated TGF-beta on MOLT-16 cells, a model intraepithelial T-cell line. Further experiments demonstrated that induction of transition coincided with a significantly increased potential for human renal epithelial cells to invade the tubular basement membrane. These data are consistent with a model in which intratubular T cells can present TGF-beta and directly induce adjacent tubular epithelial cells to transform to proliferating fibroblasts that migrate across the tubular basement membrane, producing fibrotic lesions within the renal interstitium. PMID- 14747384 TI - Paracetamol-induced renal tubular injury: a role for ER stress. AB - Paracetamol (also known as acetaminophen) causes acute and chronic renal failure. While the mechanisms leading to hepatic injury have been extensively studied, the molecular mechanisms of paracetamol-induced nephrotoxicity are poorly defined. Paracetamol induced cell death with features of apoptosis in murine proximal tubular epithelial cells. While paracetamol increased the expression of the death receptor Fas on the cell surface, the Fas pathway was not involved in the paracetamol-induced apoptosis of tubular cells. The mitochondrial pathway was not activated during paracetamol-induced apoptosis; there was no dissipation of mitochondrial potential or release of apoptogenic factors such as cytochrome c or Smac/DIABLO. However, paracetamol-induced apoptosis is a caspase-dependent process that involves activation of caspase-9 and caspase-3 in the absence of cytosolic cytochrome c or Smac/DIABLO. The authors also detected induction of endoplasmic reticulum (ER) stress, characterized by GADD153 upregulation and translocation to the nucleus, as well as caspase-12 cleavage. Interestingly, after treatment of murine tubular cells with paracetamol and calpain inhibitors, the caspase-12 cleavage product was still detectable, and calpain inhibitors were unable to protect tubular cells from paracetamol-induced apoptosis. The results suggest that induction of apoptosis may underlie the nephrotoxic potential of paracetamol and identify ER stress as a therapeutic target in nephrotoxicity. PMID- 14747386 TI - Anti-erythropoietin antibodies and pure red cell aplasia. PMID- 14747387 TI - The value of N-acetylcysteine in the prevention of radiocontrast agent-induced nephropathy seems questionable. AB - Prevention of contrast agent-induced nephropathy is of crucial importance for a number of diagnostic studies. N-Acetylcysteine (NAC) was recently reported to decrease serum creatinine levels in this setting, and its administration before radiocontrast medium administration has been widely recommended. The objective of this prospective study was to investigate whether there are effects of NAC on serum creatinine levels that are independent of alterations in GFR. Volunteers with normal renal function who did not receive radiocontrast medium were studied. Fifty healthy volunteers completed the study protocol. NAC was administered orally at a dose of 600 mg every 12 h, for a total of four doses. Surrogate markers of renal function, such as serum creatinine, urea, albumin, and cystatin C levels, were measured and estimated GFR (eGFR) was assessed immediately before the administration of NAC and 4 and 48 h after the last dose. There was a significant decrease in the mean serum creatinine concentration (P < 0.05) and a significant increase in the eGFR (P < 0.02) 4 h after the last dose of NAC. The cystatin C concentrations did not change significantly. In several studies, a protective effect of NAC on renal function after radiocontrast medium administration has been postulated. This is the first study to demonstrate an effect of NAC on creatinine levels and eGFR, surrogate markers of renal injury, without any effect on cystatin C levels. Before renoprotective effects of NAC against contrast agent-induced nephropathy are considered, the direct effects of NAC on creatinine levels, urea levels, and eGFR should be assessed. PMID- 14747388 TI - The number, quality, and coverage of randomized controlled trials in nephrology. AB - Randomized controlled trials (RCT) are the optimal study design to answer intervention questions. The authors evaluated the number, quality, and coverage of RCT in nephrology. MEDLINE was searched using the relevant medical subject headings for nephrology and 12 major specialties in internal medicine, limited by "randomized controlled trial" as a publication type. A random selection of 160 RCT in nephrology (40 for each decade) published since 1966 and an additional 270 RCT from ongoing or published Cochrane systematic reviews in various areas of nephrology, dialysis, and transplantation were evaluated for quality of reporting using standard criteria. The number of RCT published in nephrology from 1966 to 2002 (2779) is fewer than all other specialties of internal medicine (range: 5335 in hematology to 27109 in cardiology) with the proportion of all citations which are RCT being the third lowest (1.15%). There has been an increase in both indices from 1966 to 1996, but not at a greater rate than other specialties, and there has been no increase over the past 5 yr. Some areas of nephrology, in particular glomerulonephritis, are clear outliers with very low numbers of RCT to guide clinical decision-making. Overall the quality of RCT reporting in nephrology is low and has not improved over the past 30 yr with unclear allocation concealment (89%), lack of reported blinding of outcome assessors (92%), and failure to perform "intention-to-treat analysis" (50%) particularly frequent. The challenges of improving the quality and quantity of trials in nephrology are substantial, but they can be overcome by using standard guidelines and checklists for trial reporting, greater attention to the trial methods and not just the results, involving experts in trial design and reporting, multicenter collaboration, and larger and simpler trials. PMID- 14747389 TI - Randomized trial of folic acid for prevention of cardiovascular events in end stage renal disease. AB - High serum total homocysteine (tHcy) is gaining scrutiny as a risk factor for cardiovascular disease in the general population. The relationship between tHcy and mortality and cardiovascular events in patients with end-stage renal disease (ESRD) is unsettled. This randomized trial evaluates the efficacy of high-dose folic acid in preventing events in ESRD. A total of 510 patients on chronic dialysis were randomized to 1, 5, or 15 mg of folic acid contained in a renal multivitamin with a median follow-up of 24 mo. Mortality, cardiovascular events, and homocysteine levels were assessed. There were 189 deaths, and 121 patients experienced at least one cardiovascular event. Composite rates of mortality and cardiovascular events among the folic acid groups did not differ (at 24 mo: 43.7% in 1 mg group, 38.6% in 5 mg group, 47.1% in 15 mg group; log-rank P = 0.47). Unexpectedly, high baseline tHcy was associated with lower event rates. From lowest to highest quartile, event rates at 24 mo were 54.5% for Q1, 41.8% for Q2, 41.2% for Q3, and 34.7% for Q4 (log-rank P = 0.033). In contrast to some studies describing tHcy as a risk factor for mortality and cardiovascular events, this study found a reverse relationship between tHcy and events in ESRD patients. Administration of high-dose folic acid did not affect event rates. PMID- 14747390 TI - Postoperative mortality after nontraumatic lower extremity amputation in patients with renal insufficiency. AB - Mortality rates after lower extremity amputation are extremely high among dialysis patients. However, the impact of milder degrees of renal insufficiency on death rates after lower extremity amputation has not been carefully examined. In this study, the authors used data from the Department of Veterans Affairs' National Surgical Quality Improvement Program (NSQIP) to measure the association between renal dysfunction and 30-d mortality after nontraumatic amputation adjusted for confounders. The study population consisted of 16,994 patients undergoing their first NSQIP recorded amputation from January 1, 1994 through September 30, 2001. Thirty-five percent of all cohort patients had at least moderate renal insufficiency, and 52% of all postoperative deaths occurred in this group. Postoperative mortality was 9% in patients with moderate renal insufficiency, 15% in patients with severe renal insufficiency, and 16% in dialysis patients, compared with 6% in patients with normal or mildly reduced renal function. Renal insufficiency remained associated with death after adjustment for confounders (adjusted odds ratio [OR] 3.36, 95% confidence interval [CI] 2.75 to 4.10 [dialysis patients]; OR 2.54, CI 2.06 to 3.14 [severe renal insufficiency]; and OR 1.52, CI 1.32 to 1.76 [moderate renal insufficiency]). In conclusion, even moderate renal insufficiency is independently associated with postoperative death after lower extremity amputation. This finding highlights the need for a targeted approach to improving the care of patients with renal insufficiency undergoing lower extremity amputation. PMID- 14747391 TI - Analysis of the relationship between norepinephrine and asymmetric dimethyl arginine levels among patients with end-stage renal disease. AB - High sympathetic activity and alterations in nitric oxide synthesis attributable to accumulation of the endogenous nitric oxide synthase inhibitor asymmetric dimethylarginine (ADMA) have recently been identified as potential causal mechanisms for the high cardiovascular mortality rates among patients with ESRD. The link between these risk factors has not been studied. Therefore, the relationship between plasma norepinephrine (NE) and ADMA levels was examined in a large cohort of hemodialysis patients (n = 224), and whether these factors interacted in predicting all-cause mortality and new cardiovascular event rates among those patients was investigated. Plasma ADMA levels were strongly associated with plasma NE levels (P < 0.001) and to a lesser extent with heart rate (P < 0.01). In multivariate analyses, the ADMA-NE correlation was observed to be independent of age, gender, serum albumin levels, arterial pressure and antihypertensive treatment, duration of dialysis treatment, diabetes mellitus, and other risk factors. NE was an independent significant predictor of both death and cardiovascular events in Cox models not including ADMA. However, when ADMA was introduced into those models, NE became a largely nonsignificant predictor of those outcomes, whereas plasma ADMA levels emerged as a highly significant predictor of both death (P < 0.001) and cardiovascular events (P < 0.001). These findings suggest that ADMA is an intervening factor in the causal pathway leading to those outcomes. Plasma NE and ADMA concentrations are strongly related among patients with ESRD. These two factors are likely to be involved in the same causal pathway leading to death and cardiovascular events among those patients. PMID- 14747392 TI - A low, rather than a high, total plasma homocysteine is an indicator of poor outcome in hemodialysis patients. AB - An increased level of total plasma homocysteine (tHcy) is a risk factor for poor cardiovascular outcome in the general population. However, a decreased, rather than an increased, tHcy concentration may predict poor outcome in maintenance hemodialysis (MHD) patients, a phenomenon referred to as reverse epidemiology. Associations were examined between tHcy level and markers of malnutrition inflammation complex syndrome and 12-mo prospective hospitalization and mortality in 367 MHD patients, aged 54.5 +/- 14.7 (mean +/- SD) years, who included 199 men and 55% individuals with diabetes. tHcy was 24.4 +/- 11.8 micro mol/L, and 94% of the patients had hyperhomocysteinemia (tHcy >13.5 micro mol/L). tHcy had weak to moderate but statistically significant bivariate and multivariate correlations with some laboratory markers of nutrition (serum albumin, prealbumin, creatinine, and urea nitrogen) but no significant correlation with serum C-reactive protein or two proinflammatory cytokines (IL-6 and TNF-alpha). During 12 mo of follow-up, 191 MHD patients were hospitalized, 37 died, nine underwent renal transplantation, and 38 transferred out. Hospitalization rates were significantly higher in patients with lower tHcy levels. Mortality rate in the lowest tHcy quartile (17.4%) was significantly higher compared with other three quartiles (6.5 to 9.8%; Kaplan-Meier P = 0.04). Relative risk of death for the lowest tHcy quartile, even after adjustment for case-mix and serum albumin, was 2.27 (95% confidence interval, 1.14 to 4.53; P = 0.02). Hence, tHcy may be a more exclusive nutritional marker in MHD patients with no association with inflammatory measures. Despite a very high prevalence of hyperhomocysteinemia in MHD patients, lower values of tHcy are paradoxically associated with increased hospitalization and mortality. The lowest tHcy quartile confers a twofold increase in risk of death independent of hypoalbuminemia. The nutritional feature of tHcy in MHD patients may explain its reverse association with outcome. PMID- 14747393 TI - Effect of nosocomial bloodstream infection on the outcome of critically ill patients with acute renal failure treated with renal replacement therapy. AB - Critically ill patients with acute renal failure (ARF) treated with renal replacement therapy (RRT) have a high mortality. The authors evaluated a cohort of 704 consecutive intensive care unit (ICU) patients with ARF treated with RRT to determine whether there was an increased incidence of nosocomial bloodstream infection and whether this resulted in a worse outcome. The incidence of nosocomial bloodstream infection was 8.8%, higher than that reported in other series of general ICU patients and also higher than the 3.5% incidence of bloodstream infection in non-ARF patients in the same unit (P < 0.001). There were more bloodstream infections caused by Gram-positive species compared with Gram-negative species or fungi. The distribution over the species was comparable to that reported by others for a general ICU population. The outcome was evaluated with matched cohort analysis. With this technique, patients with bloodstream infection (exposed) were closely matched with patients without bloodstream infection (non-exposed) in a 1:2 ratio. Matching was based on the APACHE II system and length of stay before bloodstream infection (exposure time). Length of stay and mortality were equal in exposed and non-exposed patients. There was also no difference in hospital costs. It can be concluded that critically ill patients with ARF treated with RRT were more susceptible to nosocomial bloodstream infection. Nevertheless, the outcome was not influenced by the presence of bloodstream infection. The high mortality observed in ARF patients could therefore not be attributed to the higher incidence of bloodstream infection. PMID- 14747394 TI - The relationship between systemic and whole-body hematocrit is not constant during ultrafiltration on hemodialysis. AB - The measurement of relative blood volume (RBV) changes during ultrafiltration assume a constant mass and distribution of circulating blood components such as hematocrit. The authors examine the validity of this assumption in 10 subjects undergoing repeated direct measurements of systemic hematocrit and plasma volume (PV(icg)) using indocyanine green dilution at four stages of dialysis with intermittent ultrafiltration. Ultrasonic RBV changes were monitored. Absolute blood volumes (ABV) were initially derived for each PV(icg) estimate, and corresponding measured systemic hematocrit was adjusted by a factor of 0.86 to correct for the difference between the systemic and whole-body hematocrit (constant Fcell ratio). PV(icg) and ABV changes correlated closely (R = 0.98; P <0.001). ABV changes overestimated reduction in PV(icg) during ultrafiltration (mean difference, -140 +/- 202 ml). The calculated red cell mass however was variable (P <0.01). Fcell ratio was then adjusted at each blood volume measurement (Fcell(1), 0.87 +/- 0.02; Fcell(2), 0.89 +/- 0.03; Fcell(3), 0.94 +/- 0.06; Fcell(4), 0.94 +/- 0.04; P <0.01) to maintain a constant red cell mass (2146 +/- 460 ml). When ABV was recalculated using PV(icg), systemic hematocrit and variable Fcell (ABV(Fvariable)), the mean difference between PV(icg) changes and ABV(Fvariable) changes, was negligible (-0.2 +/- 35 ml). During intermittent ultrafiltration, RBV changes systematically underestimated the percentage reduction in ABV (mean difference, 7.7 +/- 10.6%). When corrected for variations in Fcell, ABV(Fvariable) and RBV differences were negligible (mean difference 1.2 +/- 2.6%). Varying Fcell ratio probably reflects microvascular volume change with net fluid shift from the microcirculation to macrocirculation (intravascular refill). This may result in underestimation of changes in systemic hematocrit and RBV during dialysis such that they were less than those predicted by directly measured changes in plasma volume. PMID- 14747395 TI - Increased pasma S-nitrosothiol concentrations predict cardiovascular outcomes among patients with end-stage renal disease: a prospective study. AB - The plasma concentrations of S-nitrosothiols, which are circulating nitric oxide metabolites with potential biologic activity, are increased among patients undergoing chronic hemodialysis (HD). However, the ability of S-nitrosothiols to release nitric oxide at physiologically relevant sites may be reduced among HD patients, because of impaired availability and/or activity of factors involved in S-nitrosothiol breakdown. The resultant lack of S-nitrosothiol bioavailability could contribute to the high cardiovascular risk for such patients. A possible relationship between plasma S-nitrosothiol levels and cardiac outcomes, as well as all-cause mortality rates, was investigated in a cohort of 250 chronic HD patients and who were undergoing regular dialysis three times per week were monitored for 1 yr. During that follow-up period, major cardiac events and all cause deaths were prospectively recorded. At baseline, high plasma S-nitrosothiol levels (>2 micro M, corresponding to the top quartile of all measured values) were independently associated with pulse pressure in an adjusted multivariate analysis (odds ratio, 1.03; 95% confidence interval, 1.01 to 1.05; P = 0.007). During the follow-up period, 36 patients died (16 as a result of cardiac causes) and 33 patients experienced major adverse cardiac events. In an adjusted Cox proportional-hazards model, high plasma S-nitrosothiol concentrations (i.e., the top quartile versus the three other quartiles) were an independent predictor of cardiac events (hazard ratio, 3.30; 95% confidence interval, 1.61 to 6.76; P = 0.001) but not of all-cause death. Therefore, among chronic HD patients, markedly elevated plasma S-nitrosothiol levels are associated with pulse pressure and predict cardiovascular outcomes. These findings support the hypothesis that impaired S-nitrosothiol bioavailability in uremia is an important factor for the excessive cardiovascular risk among HD patients. PMID- 14747396 TI - Vascular access and all-cause mortality: a propensity score analysis. AB - The native arteriovenous fistula (AVF) is the preferred vascular access because of its longevity and its lower rates of infection and intervention. Recent studies suggest that the AVF may offer a survival advantage. Because these data were derived from observational studies, they are prone to potential bias. The use of propensity scores offers an additional method to reduce bias resulting from nonrandomized treatment assignment. Adult (age 18 yr or more) patients who commenced hemodialysis in Australia and New Zealand on April 1, 1999, until March 31, 2002, were studied by using the Australian and New Zealand Dialysis and Transplant Association (ANZDATA) Registry. Cox regression was used to determine the effect of access type on total mortality. Propensity scores were calculated and used both as a controlling variable in the multivariable model and to construct matched cohorts. The catheter analysis was stratified by dialysis duration at entry to ANZDATA to satisfy the proportional-hazard assumption. There were 612 deaths in 3749 patients (median follow-up, 1.07 yr). After adjustment for confounding factors and propensity scores, catheter use was predictive of mortality. Patients with arteriovenous grafts (AVG) also had a significantly increased risk of death. Effect estimates were also consistent in the smaller propensity score-matched cohorts. Both AVG and catheter use in incident hemodialysis patients are associated with significant excess of total mortality. Reducing catheter use and increasing the proportion of patients commencing hemodialysis with a mature AVF remain important clinical objectives. PMID- 14747397 TI - A randomized, controlled trial of albumin versus saline for the treatment of intradialytic hypotension. AB - Intradialytic hypotension (IDH) is the most common complication of hemodialysis. Symptomatic IDH requires the administration of fluid and often results in the early termination of dialysis, both of which may prevent adequate fluid removal. The optimal fluid for the treatment of IDH remains unknown. A randomized, double blind, crossover trial was performed in 72 chronic hemodialysis patients to determine whether 5% albumin was more effective than normal saline for the treatment of IDH. The primary outcome measure was the percentage of target ultrafiltration achieved, which was defined as the actual ultrafiltration volume divided by target ultrafiltration volume. Secondary outcome measures included postdialysis BP, time to restore BP, recurrent IDH, and treatment failure (inability to restore BP with 750 ml of study fluid). The percentage of target ultrafiltration achieved was 0.84 +/- 0.17 for 5% albumin compared with 0.80 +/- 0.16 for saline (P = 0.14). The postdialysis systolic BP (121 +/- 19 mmHg versus 117 +/- 19 mmHg; P = 0.32), postdialysis diastolic BP (63 +/- 9 mmHg versus 61 +/ 9 mmHg; P = 0.33), volume of study fluid used to treat IDH (403 +/- 170 ml versus 428 +/- 191 ml; P = 0.34), time required to restore the BP (7.9 +/- 6.6 min versus 9.9 +/- 7.5 min; P = 0.09), total nursing time required to manage the hypotensive episode (15.1 +/- 7.2 min versus 15.9 +/- 7.3 min; P = 0.47), number of treatment failures (22% versus 24%; P = 1.0), and the frequency of recurrent IDH (36% versus 36%) were not significantly different when 5% albumin was used compared with saline. It is concluded that 5% albumin is no more effective than normal saline for the treatment of IDH in chronic hemodialysis patients. Normal saline should be used as the initial fluid for the treatment of IDH. PMID- 14747398 TI - Chymase-like angiotensin II-generating activity in end-stage human autosomal dominant polycystic kidney disease. AB - Autosomal dominant polycystic kidney disease (ADPKD) is characterized by exuberant inflammation and fibrosis, a process believed to contribute to progressive loss of normal renal function. Despite early-onset hypertension and intrarenal renin/angiotensin II (AngII) activation, angiotensin-converting enzyme (ACE) inhibition does not consistently confer renal protection in ADPKD. The hypothesis was that mast cells within the inflammatory interstitium release chymase, an enzyme capable of efficient conversion of AngI to AngII, providing an ACE-independent route of AngII generation. End-stage ADPKD renal tissue extracts and cyst fluids were assayed for time-dependent, chymostatin-inhibitable conversion of (125)I-AngI to (125)I-AngII under conditions of ACE and aminopeptidase inhibition by means of HPLC. Thirteen of 14 ADPKD kidney extracts exhibited chymase-like AngII-generating capacity; calculated initial reaction rates averaged 3.9 +/- 2.9 fmol AngII/min/ micro g protein with a mean maximal conversion of 55% +/- 30% of added substrate. AngII-generating activity was both protein and substrate dependent. All five cyst fluid samples were negative. Chymase-like activity was detectable in only three of six non-ADPKD kidney extracts. Immunoreactive chymase protein was present in/around mast cells within the fibrotic renal interstitium in all samples. Findings demonstrate for the first time the presence of mast cells, mast cell-associated immunoreactive chymase protein, and chymase-like AngII generating capacity in ADPKD cystic kidneys. Results support the potential for ACE-independent AngII generation and for mast cell-initiated inflammatory processes in ADPKD, each with therapeutic implications for ADPKD renal progression. PMID- 14747399 TI - Reclassification of lupus glomerulonephritis: back to the future. PMID- 14747400 TI - Albumin signals the coming of age of proteinuric nephropathy. PMID- 14747401 TI - Clinical trial in nephrology at hard end point? PMID- 14747403 TI - Calcium responses of chicken trigeminal ganglion neurons to methyl anthranilate and capsaicin. AB - Using digital fluorescence imaging, we determined the effects of methyl anthranilate (MA), an avian irritant, and capsaicin (CAP), a mammalian irritant, on intracellular calcium ([Ca(2+)](i)) in chicken trigeminal neurons. Concentration-response functions indicated that the threshold for inducing increases in [Ca(2+)](i) was higher for CAP (30 micromol l(-1)) than for MA (10 micromol l(-1)). The maximum magnitudes of [Ca(2+)](i) in response to MA and CAP were compared after normalization to 40 mmol l(-1) KCl. At equal concentrations (300 micro mol l(-1)), trigeminal neurons responded with a greater change in [Ca(2+)](i) to MA (78% of KCl) than to CAP (43% of KCl). Furthermore, at 300 micromol l(-1), 48% of neurons responded to MA whereas only 16% responded to CAP. The increases in [Ca(2+)](i) induced by both MA and CAP were dependent upon extracellular calcium. While the calcium responses to MA were also dependent on extracellular sodium, responses to CAP were not. There were separate but overlapping populations of neurons sensitive to MA and CAP. Taken together, the higher threshold concentration of CAP, the higher response magnitude to MA than CAP and the greater number of neurons sensitive to MA than CAP provide a rationale for the observed behavioral differences of birds to these two compounds. Finally, the findings that the calcium responses to MA and CAP have different ion dependencies and that there are separate populations sensitive to these compounds suggest different transduction mechanisms mediating chicken trigeminal responses to MA and CAP. PMID- 14747404 TI - Temperature regulation in burying beetles (Nicrophorus spp.: Coleoptera: Silphidae): effects of body size, morphology and environmental temperature. AB - This study compares the thermoregulatory ability of three species of burying beetle (Coleoptera: Silphidae: Nicrophorus hybridus, Nicrophorus guttula and Nicrophorus investigator) that vary significantly in body size. It also explores possible mechanisms for temperature regulation in burying beetles, including physiological and behavioral thermoregulatory strategies, and the influence of environmental temperatures on body temperature and activity times. We measured beetle thoracic and abdominal temperatures before and after short (<5 s) flights, and thoracic temperature during sustained, tethered flights and following flight in the field. We calculated two measures of thermoregulatory ability: the slope of post-flight thoracic temperature against ambient air temperature and the slope of post-flight thoracic temperature against operative flight temperature. Thoracic temperatures following flight were significantly higher than abdominal temperatures, and the largest species, N. hybridus, was determined to be the better thermoregulator, with regression slopes closer to zero (0.315-0.370) than N. guttula (0.636-0.771) or N. investigator (0.575-0.610). We also examined the roles that insulation, wing loading, physiological heat transfer, basking and perceived environmental temperature play on temperature regulation and activity times in Nicrophorus: This study shows that body size, morphological features, such as wing loading and insulation, and perceived environmental temperatures affect thermoregulation and activity times in burying beetles. PMID- 14747405 TI - The fracture properties and mechanical design of human fingernails. AB - Fingernails are a characteristic feature of primates, and are composed of three layers of the fibrous composite keratin. This study examined the structure and fracture properties of human fingernails to determine how they resist bending forces while preventing fractures running longitudinally into the nail bed. Nail clippings were first torn manually to examine the preferred crack direction. Next, scissor cutting tests were carried out to compare the fracture toughness of central and outer areas in both the transverse and longitudinal direction. The fracture toughness of each of the three isolated layers was also measured in this way to determine their relative contributions to the toughness. Finally, the structure was examined by carrying out scanning electron microscopy of free fracture surfaces and polarized light microscopy of nail sections. When nails were torn, cracks were always diverted transversely, parallel to the free edge of the nail. Cutting tests showed that this occurred because the energy to cut nails transversely, at approximately 3 kJ m(-2), was about half that needed (approx. 6 kJ m(-2)) to cut them longitudinally. This anisotropy was imparted by the thick intermediate layer, which comprises long, narrow cells that are oriented transversely; the energy needed to cut this layer transversely was only a quarter of that needed to cut it longitudinally. In contrast the tile-like cells in the thinner dorsal and ventral layers showed isotropic behaviour. They probably act to increase the nail's bending strength, and as they wrap around the edge of the nail, they also help prevent cracks from forming. These results cast light on the mechanical behaviour and care of fingernails. PMID- 14747406 TI - Drosophila CG8422 encodes a functional diuretic hormone receptor. AB - Diuretic hormone 44 (DH) is a bioactive neuropeptide that mediates osmotic balance in a wide variety of insects through increases in cAMP. It is structurally similar to mammalian corticotrophin releasing factor (CRF) peptides. In the moth Manduca and the cricket Acheta, functional studies have shown that its cognate receptor (DH-R) is related to the mammalian CRF receptor. The Drosophila genome contains two genes (CG8422 and CG12370) orthologous to Manduca and Acheta DH-Rs. Here, we present multiple lines of evidence to support the hypothesis that the orphan CG8422 G-protein-coupled receptor is a functional DH R. When expressed in mammalian cells, CG8422 conferred selective sensitivity to DH, as indicated by translocation of a beta-arrestin-2-GFP reporter from the cytoplasm to the cell membrane. Consistent with its in vivo activities in other insects, DH activation of CG8422 elicited increases in a cAMP reporter system (CRE-luciferase), with an EC(50) of 1.7 nmol l(-1). CG8422 activation by DH also led to increases in intracellular calcium but at substantially higher doses (EC(50) approximately 300 nmol l(-1)). By microarray analysis, the CG8422 transcript was detectable in Drosophila head mRNA of different genotypes and under different environmental conditions. The identification of a Drosophila receptor for the DH neuropeptide provides a basis for genetic analysis of this critical factor's roles in maintaining physiological homeostasis. PMID- 14747407 TI - 'Futile cycle' enzymes in the flight muscles of North American bumblebees. AB - In the flight muscles of European bumblebees, high activities of fructose-1,6 bisphosphatase (FbPase) relative to phosphofructokinase (PFK) have suggested a thermogenic 'futile cycle' important for regional endothermy. We find generally low activities of FbPase (0.7-19.7 units g(-1) thorax) in North American Bombus species, with the exception of Bombus rufocinctus, where activity (43.1 units g( 1) thorax) is comparable with that of European congeners. These data, taken with estimates of maximal rates of heat production by cycling, do not support a significant thermogenic role for the PFK/FbPase cycle. In agreement with earlier studies, both PFK and FbPase activities were found to scale allometrically with body size (allometric exponents -0.18 and -1.33, respectively). The cycle may serve to supplement thermogenesis or amplify glycolytic flux in rest-to-flight transitions, especially in smaller bees. PMID- 14747408 TI - Why is it worth flying at dusk for aquatic insects? Polarotactic water detection is easiest at low solar elevations. AB - Using 180 degrees field-of-view imaging polarimetry, we measured the reflection polarization patterns of two artificial surfaces (water-dummies) in the red, green and blue spectral ranges under clear and partly cloudy skies at different solar elevations. The dummies consisted of a horizontal glass pane with a matt black or matt light grey cloth underneath, imitating a dark or bright water body, respectively. Assuming that polarotactic water insects interpret a surface as representing water if the degree of linear polarization of reflected light is higher than a threshold and the deviation of the direction of polarization from the horizontal is lower than a threshold, we calculated the proportion, P, of the artificial surfaces detected polarotactically as water. We found that at sunrise and sunset P is maximal for both water-dummies and their reflection polarizational characteristics are most similar. From this, we conclude that polarotactic water detection is easiest at low solar elevations, because the risk that a polarotactic insect will be unable to recognize the surface of a dark or bright water body is minimal. This partly explains why many aquatic insect species usually fly en masse at dusk. The daily change in the reflection polarization pattern of water surfaces is an important visual ecological factor that may contribute to the preference of the twilight period for habitat searching by polarotactic water insects. Air temperature at sunrise is generally low, so dusk is the optimal period for polarotactic aquatic insects to seek new habitats. PMID- 14747409 TI - A hierarchical analysis of the scaling of force and power production by dragonfly flight motors. AB - Maximum isometric force output by single muscles has long been known to be proportional to muscle mass(0.67), i.e to muscle cross-sectional area. However, locomotion often requires a different muscle contraction regime than that used under isometric conditions. Moreover, lever mechanisms generally affect the force outputs of muscle-limb linkages, which is one reason why the scaling of net force output by intact musculoskeletal systems can differ from mass(0.67). Indeed, several studies have demonstrated that force output by intact musculoskeletal systems and non-biological systems is proportional to motor mass(1.0). Here we trace the mechanisms that cause dragonflies to achieve a change from muscle mass(0.67) scaling of maximum force output by single flight muscles to mass(1.0) scaling of dynamic force output by the intact dragonfly flight motor. In eight species of dragonflies, tetanic force output by the basalar muscle during isometric contraction scaled as muscle mass(0.67). Mean force output by the basalar muscle under dynamic conditions (workloops) that simulated in vivo maximum musculoskeletal performance was proportional to muscle mass(0.83), a significant increase in the scaling exponent over that of maximum isometric force output. The dynamic performance of the basalar muscle and the anatomy of its lever, consisting of the second moment of area of the forewing (d(2)) and the distance between the muscle apodeme and the wing fulcrum (d(1)), were used to analyze net force output by the integrated muscle-lever system (F(ind)). The scaling of d(2) conformed closely to the expected value from geometic similarity (proportional to muscle mass(0.31)), whereas d(1) scaled as muscle mass(0.54), a significant increase over the expected value from geometric similarity. F(ind) scaled as muscle mass(1.036), and this scaling exponent was not significantly different from unity or from the scaling exponent relating maximum load-lifting by flying dragonflies to their thorax mass. Thus, the combined effect of a change in the scaling of force output by the muscle during dynamic contraction compared to that during isometric contraction and the departure from geometric similarity of one of the two lever arm lengths provides an explanation for how mass(1.0) scaling of force output by the intact musculoskeletal system is accomplished. We also show that maximum muscle mass-specific net work and power output available scale as mass(0.43) and mass(0.24), respectively. PMID- 14747410 TI - Nitrogen metabolism in the African lungfish (Protopterus dolloi) aestivating in a mucus cocoon on land. AB - This study aimed to elucidate the strategies adopted by the African slender lungfish, Protopterus dolloi, to ameliorate the toxicity of ammonia during short (6 days) or long (40 days) periods of aestivation in a layer of dried mucus in open air in the laboratory. Despite decreases in rates of ammonia and urea excretion, the ammonia content in the muscle, liver, brain and gut of P. dolloi remained unchanged after 6 days of aestivation compared with the control fasted for 6 days. For specimens aestivated for 40 days, the ammonia contents in the muscle, liver and gut were significantly lower than those of the control fasted for 40 days, which suggests a decrease in the rate of ammonia production. In addition, there were significant increases in contents of alanine, aspartate and glutamate in the muscle, which suggests decreases in their catabolism. During the first 6 days and the last 34 days of aestivation, the rate of ammonia production was reduced to 26% and 28%, respectively, of the control rate (6.83 micromol day( 1) g(-1) on day 0). During the first 6 days and the next 34 days of aestivation, the averaged urea synthesis rate was 2.39-fold and 3.8-fold, respectively, greater than the value of 0.25 micromol day(-1) g(-1) for the day 0 control kept in water. No induction of activities of the ornithine-urea cycle (OUC) enzymes was observed in specimens aestivated for 6 days, because the suppression of ammonia production led to a light demand on the OUC capacity. For specimens aestivated for 40 days, the activities of carbamoyl phosphate synthetase, ornithine transcarbamylase and argininosuccinate synthetase + lyase were significantly greater than those of the control fasted for 40 days. This is in agreement with the observation that the rate of urea synthesis in the last 34 days was greater than that in the first 6 days of aestivation. P. dolloi aestivated in a thin layer of dried mucus in open air with high O(2) tension throughout the 40 days of aestivation, which could be the reason why it was able to sustain a high rate of urea synthesis despite this being an energy-intensive process. Our results indicate that a reduction in ammonia production and decreases in hepatic arginine and cranial tryptophan contents are important facets of aestivation in P. dolloi. PMID- 14747411 TI - The giant mudskipper Periophthalmodon schlosseri facilitates active NH(4)(+) excretion by increasing acid excretion and decreasing NH(3) permeability in the skin. AB - Periophthalmodon schlosseri is an amphibious and obligatory air-breathing teleost, which is extremely tolerant to environmental ammonia. It actively excretes NH(4)(+) in ammonia loading conditions. For such a mechanism to operate efficaciously the fish must be able to prevent back flux of NH(3). P. schlosseri could lower the pH of 50 volumes (w/v) of 50% seawater in an artificial burrow from pH 8.2 to pH 7.4 in 1 day, and established an ambient ammonia concentration of 10 mmol l(-1) in 8 days. It could alter the rate of titratable acid efflux in response to ambient pH. The rate of net acid efflux (H(+) excretion) in P. schlosseri was pH-dependent, increasing in the order pH 6.0<7.0<8.0<8.5. Net acid flux in neutral or alkaline pH conditions was partially inhibited by bafilomycin, indicating the possible involvement of a V-type H(+)-ATPase. P. schlosseri could also increase the rate of H(+) excretion in response to the presence of ammonia in a neutral (pH 7.0) external medium. Increased H(+) excretion in P. schlosseri occurred in the head region where active excretion of NH(4)(+) took place. This would result in high concentrations of H(+) in the boundary water layer and prevent the dissociation of NH(4)(+), thus preventing a back flux of NH(3) through the branchial epithelia. P. schlosseri probably developed such an 'environmental ammonia detoxification' capability because of its unique behavior of burrow building in the mudflats and living therein in a limited volume of water. In addition, the skin of P. schlosseri had low permeability to NH(3). Using an Ussing-type apparatus with 10 mmol l(-1) NH(4)Cl and a 1 unit pH gradient (pH 8.0 to 7.0), the skin supported only a very small flux of NH(3) (0.0095 micromol cm(-2) min(-1)). Cholesterol content (4.5 micromol g(-1)) in the skin was high, which suggests low membrane fluidity. Phosphatidylcholine, which has a stabilizing effect on membranes, constituted almost 50% of the skin phospholipids, with phosphatidyleserine and phsophatidylethanolamine contributing only 13% and 15%, respectively. More importantly, P. schlosseri increased the cholesterol level (to 5.5 micromol g(-1)) and altered the fatty acid composition (increased total saturated fatty acid content) in its skin lipid after exposure to ammonia (30 mmol l(-1) at pH 7.0) for 6 days. These changes might lead to an even lower permeability to NH(3) in the skin, and reduced back diffusion of the actively excreted NH(4)(+) as NH(3) or the net influx of exogenous NH(3), under such conditions. PMID- 14747412 TI - Female reply strategies in a duetting Australian bushcricket, Caedicia sp. (Phaneropterinae: Tettigoniidae: Orthoptera). AB - Duetting is common between the sexes of phaneropterine bushcrickets (Phaneropterinae: Tettigoniidae: Orthoptera). In this paper we describe the complex duet of an undescribed Australian species within the genus Caedicia: The male's call consists of three parts, of which the final part contains information of the species' identity and most reliably elicits the female's response. The timing of her reply usually occurs within a period of about 1 s after the male has completed his signal but may also start during the male's call. Females reply with brief clicks ranging from 1 to >10, adjusting this reply number to changes in male call duration and intensity. By using computer-synthesised calls, we discovered that the female times her reply both from cues within the male call, when she starts the reply before its conclusion, and from the end of the call, in the case where the reply follows the male call. As the number of clicks in the reply increases so the interval between clicks decreases; the female fits her entire reply within a critical time window following the male's call. The male call intensity had a marginal effect on female reply strategy. We suggest a model based on levels of female motivation, by which females may set the number of clicks in reply as well as the reply latency. PMID- 14747414 TI - A three-dimensional kinematic analysis of tongue flicking in Python molurus. AB - The forked snake tongue is a muscular organ without hard skeletal support. A functional interpretation of the variable arrangement of the intrinsic muscles along the tongue requires a quantitative analysis of the motion performance during tongue protrusion and flicking. Therefore, high-speed fluoroscopy and high speed stereo photogrammetry were used to analyse the three-dimensional shape changes of the tongue in Python molurus bivittatus (Boidae). The posterior protruding part of the tongue elongated up to 130% while the flicking anterior portion elongated maximally 60%. The differences in tongue strains relate to the absence or presence, respectively, of longitudinal muscle fibres in the peripheral tongue. Maximum overall protrusion velocity (4.3 m s(-1)) occurred initially when the tongue tip left the mouth. Maximum tongue length of approximately 0.01 body length (20 mm) was reached during the first tongue flick. These observations are discussed within the scope of the biomechanical constraints of hydrostatic tongue protrusion: a negative forward pressure gradient, longitudinal tongue compliance and axial tongue stiffness. The three dimensional deformation varied along the tongue with a mean curvature of 0.06 mm( 1) and a maximum value of 0.5 mm(-1). At the basis of the anterior forked portion of the tongue tips, extreme curvatures up to 2.0 mm(-1) were observed. These quantitative results support previously proposed inferences about a hydrostatic elongation mechanism and may serve to evaluate future dynamic models of tongue flicking. PMID- 14747413 TI - Characterization of branchial lead-calcium interaction in the freshwater rainbow trout Oncorhynchus mykiss. AB - The mechanism of branchial lead uptake and interplay with Ca(2+) transport was investigated in the freshwater rainbow trout Oncorhynchus mykiss. Lead significantly reduced Ca(2+) influx by approximately 40% and 30% after exposure to 2.3+/-0.1 and 1.4+/-0.2 micromol l(-1) dissolved lead, respectively, for 0-48 h. Acute inhibition of Ca(2+) influx by lead exhibited typical Michaelis-Menten kinetics with an approximate 16-fold increase in K(m), whereas J(max) values did not significantly change, yielding an inhibitor constant (K(i,Pb)) of 0.48 micromol l(-1). Alternative analyses suggest the possibility of a mixed competitive/non-competitive interaction at the highest lead concentration tested (4.8 micromol l(-1)). Branchial lead accumulation was reduced with increasing waterborne Ca(2+) concentrations, suggesting a protective effect of Ca(2+) against lead uptake at the gill. The apical entries of Ca(2+) and lead were both inhibited (55% and 77%, respectively) by the addition of lanthanum (1 micromol l( 1)) to the exposure water. The use of cadmium (1 micromol l(-1)) and zinc (100 micromol l(-1)) as voltage-independent calcium channel competitors also reduced branchial lead uptake by approximately 56% and 47%, respectively. Nifedipine and verapamil (up to 100 micromol l(-1)), both voltage-dependent calcium channel blockers, had no effect on gill lead accumulation. CaCl(2) injection reduced both Ca(2+) and lead uptake by the gills. This suggests transport of lead through apical voltage-independent calcium channels, similar to the entry of Ca(2+). High affinity Ca(2+)-ATPase activity was not acutely affected by lead, but a significant 80% reduction in activity occurred during exposure for 96 h to 5.5+/ 0.4 micromol l(-1) dissolved lead, indicating a possible non-competitive component to lead-induced Ca(2+) disruption. The effect of lead on Ca(2+) efflux was investigated and found to be insignificant. We conclude that uptake of lead occurs, at least in part, by the same mechanism as Ca(2+), which results in disruption of Ca(2+) influx and ultimately Ca(2+) homeostasis. PMID- 14747415 TI - Sodium and potassium currents of larval zebrafish muscle fibres. AB - The steady-state and kinetic properties of Na(+) and K(+) currents of inner (white) and outer (red) muscles of zebrafish larvae 4-6 days post-fertilization (d.p.f.) are described. In inner muscle, the outward currents were half-activated at -1.0 mV and half-inactivated at -30.4 mV, and completely inactivated within 100 ms of depolarization. The inward currents of inner fibres were half-activated at -7.3 mV and half-inactivated at -74.5 mV and completely inactivated within 5 ms of depolarization. Inner muscle fibres were found to support action potentials, while no action potentials could be evoked in outer muscles. In inner muscle fibres, all tested levels of depolarizing current above a threshold value evoked only one action potential. However, spiking at frequencies of up to 200 cycles s(-1) was evoked by the injection of depolarizing pulses separated by short hyperpolarizing currents. We suggest that the properties of the inward sodium and outward potassium currents permit high frequency firing in response to a pulsatile depolarizing input of the kind expected in fast swimming, whilst safeguarding against tetany during a strong depolarization. PMID- 14747416 TI - Swimming of larval zebrafish: ontogeny of body waves and implications for locomotory development. AB - Fish larvae, like most adult fish, undulate their bodies to propel themselves. A detailed kinematic study of the larval body wave is a prerequisite to formulate a set of functional requirements that the locomotor system must fulfil to generate the observed swimming kinematics. Lateral displacement and curvature profiles were obtained for zebrafish (Danio rerio) larvae at 2-21 days post-fertilisation for three swimming behaviours (cyclic swimming, slow starts and fast startle responses) using high-speed video. During cyclic swimming, fish larvae maintain tail beat frequencies of up to 100 Hz. The corresponding longitudinal strains, estimated from the peak curvatures of the midline, reach up to 0.19 in superficial tissue. The strain rate can reach 120 s(-1). The wave of curvature travels along the body at a near-constant rate. Posterior to the stiff head, body length-specific curvature is high and rises gently along the entire trunk to a maximum value of 6. Burst-and-coast swimming generates similar peak curvatures to cyclic swimming, but curvature rises more steeply from head to tail. Fish larvae exhibit phase shifts of 57-63 degrees between the wave of lateral displacement and the wave of curvature, resulting in a 1:1.2 ratio of body wave length to curvature wave length. During C-starts, muscle strain can reach 0.19 and superficial longitudinal strain rates approach 30 s(-1). Fish larvae do not initiate their escape response with a standing wave of curvature, although their C-starts approach a standing wave as the larvae grow older. The performance demands derived from swimming kinematics suggest that larval axial muscles have very short contraction cycles (10 ms), experience considerable strains (up to 0.2) and strain rates (up to 30 s(-1) in white muscle fibres) yet are able to power swimming for several seconds. PMID- 14747417 TI - Assessing a relationship between bone microstructure and growth rate: a fluorescent labelling study in the king penguin chick (Aptenodytes patagonicus). AB - Microstructure-function relationships remain poorly understood in primary bone tissues. The relationship between bone growth rate and bone tissue type, although documented in some species by previous works, remains somewhat unclear and controversial. We assessed this relationship in a species with extreme adaptations, the king penguin (Aptenodytes patagonicus). These birds have a peculiar growth, interrupted 3 months after hatching by the austral winter. Before this interruption, chicks undergo extremely rapid statural and ponderal growth. We recorded experimentally (by means of fluorescent labelling) the growth rate of bone tissue in four long bones (humerus, radius, femur and tibiotarsus) of four king penguin chicks during their fastest phase of growth (3-5 weeks after hatching) and identified the associated bone tissue types ('laminar', 'longitudinal', 'reticular' or 'radial' fibro-lamellar bone tissue). We found the highest bone tissue growth rate known to date, up to 171 microm day(-1) (mean 55 microm day(-1)). There was a highly significant relationship between bone tissue type and growth rate (P<10(-6)). Highest rates were obtained with the radial microarchitecture of fibro-lamellar bone, where cavities in the woven network are aligned radially. This result supports the heuristic value of a relationship between growth rate and bone primary microstructure. However, we also found that growth rates of bone tissue types vary according to the long bone considered (P<10(-5)) (e.g. growth rates were 38% lower in the radius than in the other long bones), a result that puts some restriction on the applicability of absolute growth rate values (e.g. to fossil species). The biomechanical disadvantages of accelerated bone growth are discussed in relation to the locomotor behaviour of the chicks during their first month of life. PMID- 14747419 TI - Role of stereoselective assays in bioequivalence studies of racemic drugs: have we reached a consensus? AB - The existence of stereoselectivity in metabolism and drug disposition, coupled with the existence of genetic polymorphisms and modulation of enantiomeric kinetics via special delivery systems, provides some compulsion to assess bioequivalence using stereoselective data. However, examination of the literature suggests that nonstereoselective data are commonly used for the bioequivalence assessment of drug racemates. PMID- 14747418 TI - In situ cardiac performance of Pacific bluefin tuna hearts in response to acute temperature change. AB - This study reports the cardiovascular physiology of the Pacific bluefin tuna (Thunnus orientalis) in an in situ heart preparation. The performance of the Pacific bluefin tuna heart was examined at temperatures from 30 degrees C down to 2 degrees C. Heart rates ranged from 156 beats min(-1) at 30 degrees C to 13 beats min(-1) at 2 degrees C. Maximal stroke volumes were 1.1 ml x kg(-1) at 25 degrees C and 1.3 ml x kg(-1) at 2 degrees C. Maximal cardiac outputs were 18.1 ml x kg(-1) min(-1) at 2 degrees C and 106 ml x kg(-1) min(-1) at 25 degrees C. These data indicate that cardiovascular function in the Pacific bluefin tuna exhibits a strong temperature dependence, but cardiac function is retained at temperatures colder than those tolerated by tropical tunas. The Pacific bluefin tuna's cardiac performance in the cold may be a key adaptation supporting the broad thermal niche of the bluefin tuna group in the wild. In situ data from Pacific bluefin are compared to in situ measurements of cardiac performance in yellowfin tuna and preliminary results from albacore tuna. PMID- 14747420 TI - "Cocktail" approaches and strategies in drug development: valuable tool or flawed science? AB - There is an increasing interest in the simultaneous administration of several probe substrates to characterize the activity of multiple drug-metabolizing enzymes, the so-called "cocktail" approach. However, this method remains controversial and is being investigated more extensively. No general consensus has emerged on the applicability of this approach in clinical investigation and during drug development. The objective of the article is to review this important yet specialized technique, as well as its merits, drawbacks, and potential application in drug development. Among the two-, three-, four-, five-, and six drug in vivo cocktails previously evaluated in humans, a variety of substrate probe combinations have been studied. Some probe combinations have been validated not to interact in vivo and have been useful in characterizing drug-drug interaction potential and metabolic enzyme induction in humans. For drug candidates that affect two or more in vitro pathways or are potential gene inducers, the use of a cocktail approach may facilitate the rapid delineation of the drug candidate's drug interaction potential. It may also offer the potential of providing clear guidance on safely conducting larger clinical studies and limiting comedication restrictions to only those likely to be clinically relevant. PMID- 14747421 TI - Tacrolimus dosing in adult lung transplant patients is related to cytochrome P4503A5 gene polymorphism. AB - Tacrolimus is a potent immunosuppressive agent used in lung transplantation and is a substrate for both P-glycoprotein (P-gp, encoded by the gene MDR1) and cytochrome (CYP) P4503A. A previous study by the authors identified a correlation between the tacrolimus blood level per dose with CYP3A5 and MDR1 gene polymorphisms in pediatric heart transplant patients. The objective of this study was to confirm the influence of these polymorphisms on tacrolimus dosing in adult lung transplant patients. Adult lung transplant patients who had been followed for at least 1 year after lung transplantation were studied. Tacrolimus blood level (ng/mL) per dose (mg/day) at 1, 3, 6, 9, and 12 months after transplantation was calculated as [L/D]. DNA was extracted from blood. MDR1 3435 CC, CT, and TT; MDR1 2677 GG, GT, and TT; and CYP3A5*1 (expressor) and *3 (nonexpressor) genotypes were determined by PCR amplification, direct sequencing, and sequence evaluation. Eighty-three patients were studied. At 1, 3, 6, 9, and 12 months after the transplant, a significant difference in [L/D] was found between the CYP3A5 expressor versus nonexpressor genotypes (mean +/- SD of 1.49 +/- 0.88 vs. 3.11 +/- 4.27, p = 0.01; 1.23 +/- 0.82 vs. 3.44 +/- 8.97, p = 0.05; 1.32 +/- 0.96 vs. 3.81 +/- 6.66, p = 0.005; 0.95 +/- 1.19 vs. 3.74 +/- 5.98, p = 0.0015; and 0.45 +/- 0.2 vs. 3.76 +/- 6.75, p = 0.0001, respectively). MDR1 G2677T and C3435T genotypes had only minimal effects on [L/D] at 1 and 3 months after transplantation. This study confirms the relationship of CYP3A5 polymorphisms to tacrolimus dosing in organ transplant patients. CYP3A5 expressor genotypes required a larger tacrolimus dose to achieve the same blood levels than the CYP3A5 nonexpressors at all time points during the first posttransplant year. This was not uniformly true for MDR1. The authors therefore conclude that tacrolimus dosing in adult lung transplant patients is associated with CYP3A5 gene polymorphisms. PMID- 14747422 TI - Teaching application of clinical pharmacology skills using unusual observations from clozapine overdoses. AB - Massive drug overdoses provide a unique opportunity to observe human pharmacokinetic data not otherwise ethically available. They can also provide practical examples for teaching thoughtful application of the principles of clinical pharmacology. Following a case of clozapine overdose in which onset of toxicity was delayed by 72 hours, a probable explanation was found in an exploration of three cases with unusual concentration-time profiles and revealed unexpected implications for the management of clozapine overdoses. The authors systematically addressed the possible mechanisms proposed in the literature for an unusual plateau in concentrations observed in three clozapine overdoses. The effects that the most commonly suggested explanations (i.e., delayed absorption and saturated or impaired metabolism) would have on both clozapine and norclozapine concentrations were then modeled using the data available from those three cases to provide an objective illustration for comparison. This exercise was then used as a teaching seminar, leading students through the steps required to reach a logical explanation for the observed delayed toxicity and to consider the implications for therapy. Delayed absorption best predicted the sustained serum clozapine and norclozapine concentrations observed in three cases, and modeling suggests that much of the drug remains in the gut, available for absorption for days following an overdose. As a seminar, the exercise provides students with a practical example of the value of systematically ruling out possible explanations by considering what effects various pharmacokinetic alterations would have on observed data. Absorption following massive clozapine overdose appears fundamentally different from that with conventional dosing. This suggests a potential for delayed or prolonged toxicity, extending well beyond the time frame predicted by its half-life, unless aggressive and sustained efforts are applied to remove clozapine from the gut. Data from drug overdoses provide opportunities to explore unusual aspects of pharmacokinetics, better understand future overdoses of the same agent, and present excellent material for teaching. A seminar illustrating the role that thoughtful application of pharmacologic principles had in addressing this case is now used to introduce the clinical aspects of pharmacology to students at our institutions. PMID- 14747423 TI - Questioning a class effect: does ACE inhibitor tissue penetration influence the degree of fibrinolytic balance alteration following an acute myocardial infarction? AB - There is a common belief in a class effect among angiotensin-converting enzyme (ACE) inhibitors. This is unsubstantiated for acute myocardial infarction (AMI). Because vascular tissue is a source of the endogenous fibrinolytic markers, and ACE inhibition in vascular tissue favorably influences the fibrinolytic system, the authors hypothesized that a high-tissue-penetrating ACE inhibitor would provide a more favorable reduction in plasminogen activator inhibitor-1 (PAI-1) and an increase in tissue plasminogen activator (t-PA) after AMI compared to a low-tissue-penetrating ACE inhibitor. In a randomized open-label trial, patients received the high-tissue-penetrating quinapril (n = 15) or low-tissue-penetrating enalapril (n = 15) immediately following an AMI. PAI-1 and t-PA antigen (ng/mL) were measured at baseline and through 14 days of treatment. There was no difference in baseline PAI-1 or t-PA antigen between treatments. PAI-1 antigen trended toward being lower with quinapril versus enalapril on day 1 (24.44 +/- 14.96 vs. 36.94 +/- 19.49, respectively, p = 0.059) and was significantly lower on day 3 (17.32 +/- 9.57 vs. 27.49 +/- 9.61, respectively, p = 0.009). Analysis of PAI-1 antigen over time by two-factor ANOVA with replication found significantly lower concentrations of PAI-1 antigen over the entire treatment period with quinapril versus enalapril (p < 0.003). This investigation of ACE inhibitor tissue-penetrating influence on markers of reinfarction risk suggests there may be a greater early reduction in PAI-1 with a more highly tissue penetrating ACE inhibitor. PMID- 14747424 TI - Absolute bioavailability of imatinib (Glivec) orally versus intravenous infusion. AB - The purpose of this study was to investigate the absolute bioavailability of a single oral dose of imatinib (Glivec), 400 mg (capsules vs. oral solution), compared with imatinib, 100 mg (intravenous [i.v.] infusion), in healthy subjects. Twelve subjects received a single treatment in each treatment period: a 400-mg oral dose of imatinib in capsule form or as a solution or a 100-mg i.v. infusion of imatinib. Plasma imatinib concentrations were measured following each treatment; pharmacokinetic parameters and absolute bioavailability were determined. Absolute bioavailability values (compared with i.v. infusion) for the imatinib capsule and oral solution were 98.3% and 97.2%, respectively. Both the rate and extent of imatinib absorption, as measured by C(max), partial AUC, and total AUC, were similar for the oral solution and the imatinib capsule intended for the market. The 400-mg oral dose of imatinib, as a capsule or a solution, was completely absorbed and was almost completely bioavailable (> 97%). PMID- 14747425 TI - Pharmacokinetics of intravenous, single-dose tiotropium in subjects with different degrees of renal impairment. AB - Tiotropium, a new potent anticholinergic bronchodilator, is excreted mainly by the kidney. To investigate the pharmacokinetics of tiotropium in renal impairment, the authors evaluated the pharmacokinetics and safety after administration of a single dose of intravenous tiotropium 4.8 microg, given as an infusion over 15 minutes in subjects with normal renal function and a wide range of renal impairment based on measured creatinine clearance (normal: > 80 mL/min, n = 6; mild impairment: > 50-80 mL/min, n = 5; moderate impairment: 30-50 mL/min, n = 7; severe impairment: < 30 mL/min, n =6). As expected for a drug excreted predominantly in unchanged form by the kidneys, tiotropium plasma concentrations increased as renal impairment worsened, with mean values of 55.5 (16.2 percent geometric coefficient of variation [%gCV]), 77.1 (20.1 %gCV), 101 (29.8 %gCV), and 108 (27.3 %gCV) pgh/mL for AUC(0-4h) in the normal renal function and the mild, moderate, and severe renal impairment groups, respectively. The percentage of tiotropium dose excreted unchanged in the urine decreased from 60.1% of dose (17.7 %gCV) to 59.3% (14.4 %gCV), 39.9% (34.5 %gCV), and 37.4% (10.2 %gCV) in the normal renal function and the mild, moderate, and severe renal impairment groups, respectively. Plasma protein binding of tiotropium did not significantly change in the renal-impaired subjects. Two subjects with normal renal function experienced headache 10 hours after the infusion, which was mild and transient. No adverse events occurred in subjects with renal impairment. There were no clinically relevant changes in blood pressure, pulse rate, 12-lead ECG, physical examination, hematology, or clinical chemistry, compared with baseline values, in any subject after intravenous administration of tiotropium. Tiotropium should only be used in patients with moderate to severe renal insufficiency if the potential benefit outweighs the potential risks. PMID- 14747426 TI - A single-dose, two-way crossover, bioequivalence study of dexmethylphenidate HCl with and without food in healthy subjects. AB - Attention deficit hyperactivity disorder (ADHD) in children is effectively treated by racemic oral methylphenidate (dl-MPH). The d-isomer (d-MPH) has been developed as an improved treatment for ADHD since only half the racemic dose is used. This study, performed in healthy subjects, assessed the effect of food on the pharmacokinetics of dexmethylphenidate hydrochloride (d-MPH HCl) in a single dose (2 x 10-mg tablets), two-way crossover with d-MPH administered to subjects in both a fasting state or after a high-fat breakfast. There were no serious or unexpected adverse events during the course of this study, with most events reported in comparable numbers of fed and fasted subjects. The bioequivalence of d-MPH was similar with or without food, with 90% confidence intervals of 88.2% to 104.6% and 105.9% to 118.2% for ln(C(max)) and ln[(AUC(0-infinity))], respectively. There was a marginal but statistically significant 1-hour increase in t(max) in the fed versus fasted state, reflecting an absorption delay. The rate of formation of the major metabolite, d-ritalinic acid (d-RA), was marginally decreased ( approximately 14%) after food. The extent of exposure to d RA was similar (within 1.2%) between both treatments. There was a marginal but statistically significant difference in mean t(max) for d-RA between fed and fasted conditions, with peak concentration occurring 1.5 hours later after d-MPH administration with food. There was no measurable in vivo chiral inversion of d MPH to l-MPH in plasma. In addition, the metabolism of d-MPH was stereospecific as d-MPH only produced d-RA. In summary, food had no substantial effect on the bioavailability of d-MPH, with an equivalent rate and extent of exposure obtained. Therefore, d-MPH can be administered without regard to food intake. PMID- 14747427 TI - Pharmacokinetics, tolerability, and safety of aripiprazole following multiple oral dosing in normal healthy volunteers. AB - Two 14-day, placebo-controlled, double-blind studies evaluated the fasting pharmacokinetics, safety, and tolerability of aripiprazole, a new antipsychotic, in healthy male subjects. In Study 1, 37 subjects were randomized to aripiprazole 5 mg, 10 mg, 15 mg, 20 mg, or placebo once daily. In Study 2, 11 subjects were randomized to aripiprazole, titrated from 10 to 30 mg/day, or placebo. Aripiprazole had linear pharmacokinetics over 5 to 30 mg/day, which were described by a two-compartment open model, with first-order absorption. In Study 1, mean elimination half-life ranged from 47 to 68 hours with aripiprazole, with apparent systemic clearance (CL/F) of approximately 3.45 L/h. In Study 2, mean elimination half-life was 59 hours (CL/F approximately 4.0 L/h). Adverse events were generally mild to moderate, were transient in nature, and commonly occurred within the first 3 days of dosing. Clinical laboratory assessments, electrocardiogram, electroencephalogram, and prolactin levels showed no clinically significant changes during the studies. PMID- 14747428 TI - Characterization of the penetration of garenoxacin into the breast milk of lactating women. AB - The primary objective of this study was to characterize the extent of excretion of garenoxacin, a novel des-F(6)-quinolone antimicrobial, into the breast milk of lactating women. A secondary objective was to determine the time after dose administration that garenoxacin was no longer detected in breast milk so as to define when a mother may resume breastfeeding if it was interrupted for garenoxacin administration. Six healthy, lactating women (age [mean +/- SD]: 32 +/- 6 years; weight: 68.3 +/- 19.8 kg; body mass index: 26 +/- 5 kg/m(2)) who had completed weaning their infants were administered a single 600-mg oral dose of garenoxacin. Plasma samples were collected predose and repeatedly up to 72 hours postdose. Breast milk was collected predose and for 6- to 12-hour intervals repeatedly up to 120 hours postdose. Breast milk/plasma concentration ratios for garenoxacin ranged from 0.35 to 0.44 up to 24 hours postdose, and the mean peak breast milk concentration was 3.0 microg/mL (0- to 6-h collection interval). Overall, garenoxacin exposure in breast milk was minimal, with a mean of 0.07% of the administered dose recovered within 120 hours. Indeed, garenoxacin was undetectable in the breast milk of a majority of subjects within 84 hours of dosing. As such, an infant nursing from a mother who had received a single 600-mg oral dose of garenoxacin could theoretically be exposed to 0.42 mg of garenoxacin (0.105 mg/kg/day for a 4-kg infant over the period of 5 days of nursing). If extrapolated to a 14-day course of garenoxacin 600 mg once daily, total exposure would be approximately 5.88 mg. These findings indicate that, like other quinolone antimicrobials, garenoxacin is secreted in breast milk. PMID- 14747429 TI - Lumiracoxib: pharmacokinetic and pharmacodynamic profile when coadministered with fluconazole in healthy subjects. AB - This two-way crossover study evaluated the effect of fluconazole on the pharmacokinetics and selective COX-2 inhibition of lumiracoxib. Thirteen healthy subjects were randomized to fluconazole (day 1: 400 mg; days 2-4: 200 mg) or no drug. On day 4, all subjects received a single dose of lumiracoxib (400 mg). Lumiracoxib pharmacokinetics were assessed during the following 48 hours. Thromboxane B(2) (TxB(2)) inhibition was measured prior to lumiracoxib dosing and 2 hours afterwards. Fluconazole caused a small (18%) but not clinically relevant increase in lumiracoxib mean AUC(0- infinity ) but had no effect on lumiracoxib mean C(max). The geometric mean ratio (lumiracoxib plus fluconazole/lumiracoxib alone) for AUC(0- infinity ) was 1.19 (90% confidence interval [CI] = 1.12, 1.27) and for C(max) was 1.11 (90% CI = 0.98, 1.27). The decrease in TxB(2) from predose was not significantly different for lumiracoxib (11.8%) or lumiracoxib plus fluconazole (7.1%); no correlation between lumiracoxib concentration and TxB(2) decrease was seen. As fluconazole is a strong inhibitor of cytochrome P450 (CYP) 2C9, other CYP2C9 inhibitors are unlikely to affect lumiracoxib pharmacokinetics with clinical relevance, making dosage adjustment unnecessary. PMID- 14747430 TI - Recipes for adult stem cell plasticity: fusion cuisine or readymade? AB - A large body of evidence supports the idea that certain adult stem cells, particularly those of bone marrow origin, can engraft at alternative locations, particularly when the recipient organ is damaged. Under strong and positive selection pressure these cells will clonally expand/differentiate, making an important contribution to tissue replacement. Similarly, bone marrow derived cells can be amplified in vitro and differentiated into many types of tissue. Despite seemingly irrefutable evidence for stem cell plasticity, a veritable chorus of detractors has emerged, some doubting its very existence, motivated perhaps by more than a little self interest. The issues that have led to this situation include the inability to reproduce certain quite startling observations, and extrapolation from the behaviour of embryonic stem cells to suggest that adult bone marrow cells simply fuse with other cells and adopt their phenotype. Although these issues need resolving and, accepting that cell fusion does appear to allow reprogramming of haemopoietic cells in special circumstances, criticising this whole new field because some areas remain unclear is not good science. PMID- 14747431 TI - Rational requesting or rationing testing? PMID- 14747432 TI - More guidance on growth hormone deficiency. PMID- 14747433 TI - The investigation of short stature: a survey of practice in Wales and suggested practical guidelines. AB - AIM: To survey the investigation of short stature in Wales and suggest guidelines to improve practice. METHODS: Questionnaires were circulated to paediatricians and consultant clinical biochemists or consultant chemical pathologists at 13 Welsh hospitals where children with short stature are investigated. RESULTS: A 100% response was obtained from laboratory and clinical staff. Clinicians screened 1-50 patients each year (median, 10). Growth hormone (GH) deficiency was subsequently diagnosed in 0-30% (median, 10%) and GH treatment started in 30-100% (median, 100%) of patients. Five paediatricians and eight laboratories had written investigative protocols. Investigation of GH secretion was initiated in some centres before a complete clinical evaluation was carried out. Various screening tests for GH deficiency, including insulin-like growth factor 1 (IGF 1), random GH, and exercise tests were used. The clonidine stimulation test was used to assess the GH axis in most centres but eight different protocols were described. GH was measured in four Welsh laboratories using two automated immunoassay methods. However, nine different ranges of cutoff values for defining abnormal GH responses were quoted, and in three centres laboratories and paediatricians quoted different cutoffs. CONCLUSION: This survey demonstrates the need for practical guidelines for the investigation and management of short stature in children, agreed by paediatricians and their laboratory colleagues. The guidelines should encompass the initial clinical investigation, assessment of the GH-IGF-1 axis (using standardised protocols), and provision for the transition to adult management. This article presents practical guidelines based on published points for good practice. PMID- 14747434 TI - The effect of a symptom related "gating policy" on ANCA requests in routine clinical practice. AB - BACKGROUND: Most positive antineutrophil cytoplasmic antibody (ANCA) results are associated with non-vasculitic conditions, and guidelines have been proposed for the judicious use of this test. The outcome of applying similar guidelines in a routine laboratory is reported. METHODS: All immunology requests (6500) over six months were selected, and those requesting ANCA were studied for the appropriateness of the clinical data supporting the request, the presence of ANCA in those samples tested, and the final diagnosis. Antibodies were detected by indirect immunofluorescence. RESULTS: ANCA testing was requested in 287 samples. Application of a "gating policy", which refuses analysis on requests that are not supported by clinical data suggestive of systemic vasculitis, made clinicians more selective about the patients for whom they requested ANCA testing. The percentage of "appropriate" screens for systemic vasculitis was relatively high (212 of 287 requests: 72.5%). Only one of the remainder, for whom ANCA testing was initially refused, developed an ANCA related systemic vasculitis in the two years after the study, but the delay in reporting her positive ANCA was only two days. Most of the samples tested were negative (155 of 212), but most (42 of 57) of the patients with positive ANCA results were found to have a systemic vasculitis. CONCLUSIONS: A gating policy to select requests supported by clinical data suggestive of systemic vasculitis makes ANCA testing more clinically relevant and cost effective. Studies where guidelines can be proposed and their effects measured are important in the light of clinical governance and evidence based medicine. PMID- 14747435 TI - A study of public opinion on the use of tissue samples from living subjects for clinical research. AB - AIMS: To assess public opinion on the use of tissue samples from living adults and children for clinical research. METHODS: A questionnaire study of 100 healthy volunteers (100% response rate) from a Newcastle NHS dental practice. The issues investigated were the types of tissues that individuals were prepared to donate for research, the type of research donors would be prepared to consent to, and attitudes to research on children's tissues. RESULTS: Eighteen per cent of the participants said that they would not give consent for research to be carried out on their tissues, 50% would not give consent for the donation of a child's tissues. Only 26% of subjects said that they would give consent for research on genetic cloning compared with 82% for cancer research. Sex differences existed in the responses. CONCLUSIONS: Greater research attention needs to be given to public opinion on the use of tissue from living subjects for medical research to facilitate drafting of new legislation. PMID- 14747436 TI - The diagnostic value and cost effectiveness of routine fungal stains in a dermatopathology service of a district general hospital. AB - AIMS: To evaluate the use and effectiveness of fungal stains in a dermatopathology service of a district general hospital. METHODS: A retrospective analysis of skin biopsies submitted over three years, where fungal stains were used; the results were correlated with clinical history and case notes. RESULTS: In total, 99 cases were studied for fungi with the periodic acid Schiff stain with diastase. Fungi were present in seven cases; fungi had been suggested in the differential diagnosis of three of these cases but were an unexpected finding in four cases. CONCLUSION: Non-specific clinical details should prompt early fungal staining and non-specific microscopic findings or inappropriate well recognised skin reaction patterns should warrant the exclusion of fungal infection. The finding of at least one case of unexpected fungal infection is justified financially and for patient best management where clinical and microscopic findings are non-specific or inappropriate. PMID- 14747437 TI - Clinical evaluation of the COBAS Amplicor HBV monitor test for measuring serum HBV DNA and comparison with the Quantiplex branched DNA signal amplification assay in Taiwan. AB - AIMS: To evaluate the performance characteristics and clinical usefulness of the COBAS Amplicor HBV monitor (COBAS-AM) test in Taiwan and to examine its correlation with the Quantiplex branched DNA signal amplification (bDNA) assay for measuring serum hepatitis B virus (HBV) DNA concentrations. METHODS: HBV DNA was measured by the COBAS-AM test in 149 sera from chronic HBV infected patients that had previously been analysed by the bDNA assay. RESULTS: The COBAS-AM test showed good reproducibility, with acceptable intra-assay and interassay coefficients of variation (1.6% and 0.9%, respectively) and good linearity (r2=0.98). The overall sensitivity of the COBAS-AM test was significantly higher than that of the bDNA assay (95.3% v 83.2%): 69.6% of samples with HBV DNA below the detection limit of the bDNA assay could be measured by the COBAS-AM test. There was a significant correlation between the results of the two assays (r=0.901; p<0.0001). On average, the results derived from the COBAS-AM test were 0.55 log lower than those of the bDNA assay. HBV DNA concentrations were significantly higher among HBV e antigen (HBeAg) positive patients than negative ones, and higher among patients with abnormal alanine aminotransferase (ALT) concentrations than those with normal ALT concentrations (p=0.0003). CONCLUSIONS: The COBAS-AM assay, more sensitive in HBeAg negative samples than the bDNA assay, can effectively measure HBV DNA concentrations in Taiwanese patients. HBV DNA values measured by the COBAS-AM test and bDNA assay correlate significantly. PMID- 14747438 TI - Computer based receptogram approach: an objective way of assessing immunohistochemistry of androgen receptor staining and its correlation with hormonal response in metastatic carcinoma of prostate. AB - AIMS: To categorise the immunostaining heterogeneity of androgen receptors in metastatic carcinoma of the prostate using a pattern oriented approach and to correlate the results with response to hormonal treatment. METHODS: Paraffin wax embedded tumour sections from 85 patients with metastatic carcinoma of the prostate were processed for immunocytochemistry and stained for the androgen receptor using antiandrogen receptor antibodies. A computer based image analysis system was used to analyse the pattern of nuclear immunostaining in a minimum of 500 nuclei/slide. Depending on the nuclear receptor content and concentration, receptogram patterns were established for each specimen. The receptogram pattern was correlated with clinical response to hormonal treatment. RESULTS: Clinical response to hormonal treatment was documented using prostate specific antigen as the marker into responders (good, fair, stable) and non-responders. Forty four of 48 patients who responded to hormonal treatment had type 1 (35) or type 3 (nine) receptograms, which are characterised by a unimodal peak or multimodal peaks within a narrow concentration range. Thirteen of the 18 patients who stabilised had type 1 or type 3 receptograms. Seventeen of the 19 patients who did not respond to hormonal treatment had either type 2 or type 4 receptograms, which are characterised by skewed or bimodal androgen receptor distribution. Positive and negative predictive values of receptograms were 96.5% and 63%, respectively. CONCLUSIONS: Image analysis of androgen receptor immunostaining with a receptogram oriented approach provides important prognostic information that can be used to predict response to hormone treatment in patients with metastatic carcinoma of the prostate. PMID- 14747439 TI - Non-neoplastic granulosa cells within ovarian vascular channels: a rare potential diagnostic pitfall. AB - AIMS: To describe six cases seen in consultation in which artefactual vascular involvement within the ovary by benign granulosa cells caused diagnostic confusion. METHODS/RESULTS: In five cases, the initial favoured diagnoses of the submitting pathologists were metastatic carcinoma (three cases) and immature neural elements within a teratoma (two cases). In two cases, the ovary contained a benign cystic teratoma (one with struma ovarii), in two cases endometriosis, in one case follicular cysts, and in the other no pathological lesion was present. In all cases, several small ovarian vascular channels contained cohesive groups of cells with mildly atypical nuclei and cytoplasm, which varied from scant to abundant and eosinophilic. In four cases, mitotic figures were identified. The cells were morphologically consistent with benign granulosa cells and were associated in four cases with a nearby follicle lined by similar cells. There was no evidence of a mass lesion, grossly or histologically, to suggest a granulosa cell tumour. The nature of the cells was confirmed using immunohistochemistry for alpha inhibin and calretinin in one case. CONCLUSIONS: This phenomenon is probably an artefact secondary to surgical trauma or sectioning within the laboratory; alternatively, it could be related to ovulation. It is important that this benign process is not misinterpreted as cancer, either primary or metastatic, which may prompt inappropriate treatment or investigations that are not needed. PMID- 14747440 TI - Hepatic myospherulosis complicating portal vein embolisation. AB - AIMS: Myospherulosis is a rare condition characterised by sac-like structures containing spheroid bodies in cysts or cystic spaces in the tissue. This condition has not previously been reported in the liver. The association with previous portal vein embolisation using a mixture of butyl 2-cyanoacrylate and ethiodised oil and the proposed mechanism of pathogenesis are discussed. METHODS: Samples from 8 patients treated by hepatectomy after portal vein embolisation using a mixture of butyl 2-cyanoacrylate and ethiodised oil were retrieved from the archives of the United Christian Hospital, Hong Kong. The histological specimens were reviewed. A panel of histochemical and immunohistochemical stains was used. RESULTS: All cases showed hepatic myospherulosis within the veins. The veins were denuded of endothelium, which was replaced by granulation tissue and fibrous tissue with a lymphoplasmacytic infiltrate. Foreign body-type giant cells (six cases) and eosinophilic infiltrates (seven cases) were noted in most cases. Both parent bodies and endobodies were stained red by Papanicolaou and Masson's trichrome and stained blue by solochrome cyanine. The endobodies showed immunoreactivity towards glycophorin A. They were negative for Alcian blue, periodic acid Schiff, Grocott, and Ziehl-Neelsen stains. CONCLUSIONS: The endobodies of myospherulosis may be misdiagnosed as fungi or algae by the unwary. The clinical history, intravascular location, lack of staining with periodic acid Schiff and Grocott stains, and positive glycophorin A staining are generally sufficient for a confident diagnosis of myospherulosis. PMID- 14747441 TI - Rapid, accurate genotyping of the common -alpha(4.2) thalassaemia deletion based on the use of denaturing HPLC. AB - AIMS: To develop an alternative assay for specific genotyping of the -alpha(4.2) thalassaemia deletion based on the DNA sequence features surrounding the breakpoint. METHODS: The 5' and 3' ends of the breakpoint regions of the alpha(4.2) allele and the normal homologous segments were sequenced in Chinese individuals. A sequence haplotype composed of four single nucleotide variations within the X2/X1 box of the -alpha(4.2) breakpoint region was found in all of the 10 Chinese -alpha(4.2) thalassaemia alleles studied. Based on these findings, a novel polymerase chain reaction (PCR)/denaturing high performance liquid chromatography (DHPLC) assay was developed for rapid genotyping of the alpha(4.2) allele instead of traditional Southern blotting or Gap-PCR. This method involves amplification of the alpha globin target sequence encompassing these four polymorphic sites, followed by a partially denaturing HPLC analysis using the transgenomic WAVE DNA fragment analysis system. RESULTS: The three major genotypes (-alpha4.2/alphaalpha, -alpha(4.2)/--SEA, and alphaalpha/alphaalpha) could be distinguished through the characteristic chromatograms generated by the WAVE system. The accuracy of this technique was evaluated blindly, and the results were 100% (40 of 40) concordant with the genotypes previously characterised by Southern blotting or Gap-PCR. CONCLUSIONS: This study validates the PCR/DHPLC approach as a simple, rapid, highly accurate, and cost effective method, potentially adaptable for use in epidemiological surveys, genetic screening, and diagnosis of silent alpha+ thalassaemia and Hb H disease. PMID- 14747442 TI - Alpha0 thalassaemia as a result of a novel 11.1 kb deletion eliminating both of the duplicated alpha globin genes. AB - AIMS: To characterise a novel 11.1 kb deletion that eliminated both of the duplicated alpha globin genes, giving rise to a typical alpha0 thalassaemia phenotype in four carriers from a Chinese family. METHODS: Haematological investigations were carried out on all family members. The seven common forms of alpha thalassaemia were screened for by the polymerase chain reaction (PCR) and Southern blotting was used to analyse the alpha globin gene cluster. DNA sequence analysis of the entire alpha1 and alpha1 globin gene region was carried out and reverse transcription (RT)-PCR was used to investigate the transcription levels of the alpha and beta globin genes. RESULTS: The breakpoints were found to lie between coordinates 31695-31724 and 42846-42867 of the alpha globin gene cluster (NG_000006), with a total of about 11,135 nucleotides deleted. These sequences are involved in (CA)n repeats, suggesting a homologous recombination event. RT PCR analysis gave a transcription level of the alpha globin gene in heterozygotes comparable with that of SEA deletion heterozygotes, confirming no output of alpha globin from the linked pair of alpha globin genes. The heterozygosity for this novel deletion was confirmed by PCR diagnosis in all four carriers from this family. CONCLUSIONS: This rare mutation constitutes an additional heterogeneous defect causing alpha thalassaemia in the Chinese population. PMID- 14747443 TI - The immunohistochemical localisation of somatostatin receptors 1, 2, 3, and 5 in acoustic neuromas. AB - AIMS: Acoustic neuroma is a benign tumour, which develops through an overproliferation of Schwann cells along the vestibular nerve. Somatostatin is a naturally occurring peptide, which exerts antiproliferative and antiangiogenic effects via five membrane bound receptor subtypes. The aim of this study was to determine whether somatostatin receptor subtypes (SSTRs) 1, 2, 3, and 5 are present in acoustic neuromas. METHODS: The expression of SSTRs 1, 2, 3, and 5 was studied in both the Schwann cells and blood vessels of eight acoustic neuroma specimens, by means of immunohistochemistry using novel rabbit polyclonal antibodies raised against human SSTR 1, 2, and 5 subtype specific peptides, and a commercial anti-SSTR3 antibody. RESULTS: SSTR2 was the most prevalent subtype in Schwann cells (seven of eight), with intermediate expression of SSTR3 (six of eight), and lower expression of SSTRs 1 and 5 (four of eight and five of eight, respectively). There was ubiquitous vascular expression of SSTR2, with no evidence of SSTR 1, 3, or 5 expression in blood vessels. CONCLUSION: SSTRs 1, 2, 3, and 5 are differentially expressed in acoustic neuromas. Somatostatin analogues may have a therapeutic role in the management of this rare and challenging condition. PMID- 14747444 TI - The immunohistochemical expression of calcitonin receptor-like receptor (CRLR) in human gliomas. AB - BACKGROUND: Gliomas are the most common primary tumours of the central nervous system and exhibit rapid growth that is associated with neovascularisation. Adrenomedullin is an important tumour survival factor in human carcinogenesis. It has growth promoting effects on gliomas, and blockade of its actions has been experimentally shown to reduce the growth of glioma tissues and cell lines. There is some evidence that the calcitonin receptor-like receptor (CRLR) mediates the tumorigenic actions of adrenomedullin. AIM: To determine whether CRLR is expressed in human gliomas and the probable cellular targets of adrenomedullin. METHODS: Biopsies from 95 human gliomas of varying grade were processed for immunohistochemical analysis using a previously developed and characterised antibody to CRLR. RESULTS: All tumour specimens were positive for CRLR. As previously found in normal peripheral tissues, CRLR immunostaining was particularly intense in the endothelial cells. This was evident in all the various vascular conformations that were observed, and which are typical of gliomas. In addition, clear immunostaining of tumour cells with astrocyte morphology was observed. These were preferentially localised around vessels. CONCLUSIONS: This study has shown for the first time that the CRLR protein is present in human glioma tissue. The expression of the receptor in endothelial cells and in astrocytic tumour cells is consistent with the evidence that its endogenous ligand, adrenomedullin, may influence glioma growth by means of both direct mitogenic and indirect angiogenic effects. CRLR may be a valuable target for effective therapeutic intervention in these malignant tumours. PMID- 14747445 TI - A follow up model for patients with atrophic chronic gastritis and intestinal metaplasia. AB - AIM: To devise a follow up model for patients with gastric cancer associated lesions, such as atrophic chronic gastritis (ACG) and intestinal metaplasia (IM). METHODS: Cohort study of 144 patients, followed for a minimum of one year, in whom at least two upper gastrointestinal endoscopic biopsies in flat gastric mucosa provided a diagnosis of ACG, IM, or low grade dysplasia (LGD). RESULTS: Of those diagnosed with ACG at first endoscopic biopsy (entry biopsy), 12% progressed to LGD in outcome biopsy, as did 8% of those with type I IM, 38% with type II or III IM, and 32% with LGD. Type of IM at entry independently predicted progression to LGD and cancer. Type II and III IM had a higher rate of progression to LGD than type I IM, which showed an indolent behaviour similar to ACG. Patients with type II or III IM were at higher risk for development of dysplasia, and 7% of patients with type III IM at first biopsy progressed to high grade dysplasia (HGD), whereas no cases of ACG or type I/II IM progressed to HGD during the first three years. CONCLUSION: Patients with ACG or IM could possibly be allocated to different management schedules, based on differences in rate and proportion of progression to LGD or HGD. Less intensive follow up (two/three yearly with "serological evaluation" (pepsinogen)) may suit those with ACG or type I IM. Patients with type III IM may benefit from six to 12 monthly improved endoscopic examination (magnification chromoendoscopy). PMID- 14747446 TI - Lack of vascular adventitial fibroblastic cells in tumour stroma of intestinal type and solid-type gastric carcinomas. AB - AIMS: To investigate the roles of vascular adventitial fibroblastic cells in tumour stroma, the distribution of vascular adventitial fibroblastic cells was studied in gastric carcinomas. METHODS: In total, 50 surgically resected gastric carcinomas (43 intestinal type, and seven solid type) and their normal tissues were examined. Vascular adventitial fibroblastic cells are positive for CD34 but negative for CD31. To differentiate vascular adventitial fibroblastic cells from vascular endothelial cells, immunostaining for CD34 and CD31 was performed. Immunostaining for high molecular weight caldesmon was also performed to recognise vascular media. RESULTS: In normal gastric tissues, CD34 positive fibroblastic cells were found just outside the vascular media, namely vascular adventitial fibroblastic cells. In contrast, all of the 43 intestinal-type and seven solid-type gastric carcinomas had no vascular adventitial fibroblastic cells in the tumour stroma. CONCLUSIONS: These results suggest that a lack of vascular adventitial fibroblastic cells is associated with tumour stroma formation in intestinal-type and solid-type gastric carcinomas. PMID- 14747447 TI - Demographic study of leukaemia presenting within the first 3 months of life in the Northern Health Region of England. AB - AIMS: To determine the incidence and outcome of congenital leukaemia. METHODS: Retrospective population based study of putative leukaemia arising during the first 3 months of life over an 18 year period within the Northern Health Region of England. RESULTS: Nine infants with putative leukaemia were identified. Five had acute leukaemia and four had transient myeloproliferative disorder (TMD). Trisomy 21, either as Down's syndrome or perhaps restricted to proliferating marrow cells, was present in all four infants with TMD. The incidence of congenital acute leukaemia was 8.6/10(6) live births/year, but would be less than half this value if only patients presenting within 4 weeks of birth were counted. Remission was induced in three of the five patients with acute leukaemia. One patient, who presented at birth, remains well five years after diagnosis. All four patients with TMD survive. CONCLUSIONS: Congenital leukaemia is very rare but is not inevitably fatal. Finding trisomy 21 in spontaneously dividing blood or bone marrow cells of an infant with putative acute leukaemia, particularly within 3 months of birth, should encourage a cautious clinical approach and suggests that the diagnosis might be TMD. PMID- 14747448 TI - Gastrin releasing peptide and gastrin releasing peptide receptor expression in gastrointestinal carcinoid tumours. AB - AIMS: To establish whether gastrin releasing peptide (GRP) and the GRP receptor (GRPR) are expressed together in gastrointestinal carcinoid tumours. METHODS: Twenty six carcinoid tumours from the stomach, small intestine, appendix, and colorectum were investigated by immunohistochemistry for GRP and GRPR. RESULTS: GRP was detected in nine of 19 tumours and GRPR in 22 of 26. Coexpression of both the ligand and receptor was seen in six of 19 cases. GRPR but not GRP was more strongly expressed in appendix and colonic tumours. CONCLUSIONS: GRP and GRPR are produced by a large number of gastrointestinal carcinoid tumours. An autocrine/paracrine pathway may exist for GRP stimulated cell proliferation in some of these neoplasms, analogous to that seen in small cell anaplastic carcinoma of the lung. PMID- 14747449 TI - Agreement between preoperative core needle biopsy and postoperative invasive breast cancer histopathology is not dependent on the amount of clinical material obtained. AB - AIMS: To establish the relation between the amount of breast core needle biopsy (CNB) material examined and agreement between preoperative and postoperative histopathology parameters in invasive breast cancer. METHODS: The CNB and surgical specimen histopathology reports of 113 patients with invasive breast carcinoma were reviewed and the total amount of CNB material examined for each case was determined. Agreement was calculated for tumour type, grade, mitoses, nuclear pleomorphism, and tubule formation. Associations between the amount of CNB material and histopathology agreement before and after surgery were explored using binary logistic regression. RESULTS: Tumour type and grade agreed in 65.4% and 61.6% of cases, respectively. The components used to calculate grade--nuclear pleomorphism (57.4%), mitoses (59.4%), and tubule formation (55.6%)--agreed slightly less frequently. The proportion of cases with preoperative and postoperative assessments that agreed did not depend on the number of cores collected or the total amount of material examined. CONCLUSION: Neither tumour type and grade, nor the individual components used to calculate grade agreed consistently between the CNB and surgical specimen. The number of cores collected and the total amount of material reviewed by the pathologist does not influence the likelihood of agreement between preoperative and postoperative histopathology reports. PMID- 14747450 TI - Polymicrobial candidaemia revealed by peripheral blood smear and chromogenic medium. AB - Candida spp are the fourth most common group of nosocomial pathogens isolated from patients on medical, surgical, and intensive care wards. Polymicrobial candidaemia has rarely been described. The diagnosis of candidaemia from peripheral blood smears has not been widely reported. This report describes the case of a young woman suffering from Ewing's sarcoma who developed a syndrome of septic shock. Deep fungal infection was diagnosed from a systematic peripheral blood smear and yeasts were isolated within 24 hours. A subculture on CHROMagar Candida allowed the differentiation and presumptive identification of Candida tropicalis and Candida krusei. Species identification was confirmed by the ID 32C system. This report underlines the usefulness of peripheral blood smears in the diagnosis of fulminant deep fungal infections, and of a differential isolation medium in the rapid presumptive identification of clinically important yeast species from clinical samples. This medium is particularly useful for the detection of mixed fungal infections, allowing early and better adapted antifungal treatment. PMID- 14747451 TI - Rapid identification of Staphylococcus aureus from BacT/ALERT blood culture bottles by direct Gram stain characteristics. AB - The rapid identification of Staphylococcus aureus from positive blood cultures provides important clinical and therapeutic information. Using criteria based on direct Gram stain characteristics, an experienced microscopist was able to distinguish S aureus from other staphylococci isolated from BacT/ALERT blood culture bottles with an overall sensitivity of 89% and specificity of 98%. Furthermore, this method was readily taught to a clinical microbiologist who had not previously used the method first hand. Laboratories using the BacT/ALERT blood culture system should become familiar with these criteria so that S aureus bacteraemia can be identified rapidly. PMID- 14747452 TI - Central nervous system Aspergillus fumigatus infection after near drowning. AB - AIMS: To report the case of a 26 year old white man, who developed chronic meningitis and intracerebral granulomata 15 days after an episode of near drowning in a swamp. METHODS: Aspergillus fumigatus was isolated from cerebrospinal fluid cultures. RESULTS: The patient died 70 days after the symptoms were first noticed, and seven days after a subarachnoid haemorrhage. Aspergillus has never been reported before as a cause of intracranial infection after near drowning. CONCLUSIONS: Physicians must be aware of this possibility when confronted with such a situation, because there are now effective therapeutic options for systemic aspergillosis. PMID- 14747453 TI - Infection of the CNS by Scedosporium apiospermum after near drowning. Report of a fatal case and analysis of its confounding factors. AB - This report describes a fatal case of central nervous system pseudallescheriasis. A 32 year old white man presented with headache and meningismus 15 days after nearly drowning in a swine sewage reservoir. Computerised tomography and magnetic resonance imaging of the head revealed multiple brain granulomata, which vanished when steroid and broad spectrum antimicrobial and antifungal agents, in addition to dexamethasone, were started. Cerebrospinal fluid analysis disclosed a neutrophilic meningitis. Treatment with antibiotics and amphotericin B, together with fluconazole and later itraconazole, was ineffective. Miconazole was added through an Ommaya reservoir, but was insufficient to halt the infection. Pseudallescheria boydii was finally isolated and identified in cerebrospinal fluid cultures, a few days before death, three and a half months after the symptoms began. Diagnosis was delayed because of a reduction in the lesions after partial treatment, which prevented a stereotactic biopsy. Physicians should be aware of this condition, and provide prompt stereotactic biopsy. Confirmed cases should perhaps be treated with voriconazole, probably the most effective, currently available treatment for this agent. PMID- 14747454 TI - Waterhouse-Friderichsen syndrome as a result of non-meningococcal infection. AB - Waterhouse-Friderichsen syndrome--massive adrenal haemorrhage in the setting of overwhelming clinical sepsis--is usually taken at necropsy to indicate meningococcal infection, and may be the only evidence of this pathogen. This report describes three fatal cases of the syndrome in which the causative organism proved to be a streptococcus. The organisms were detected during routine coroners' autopsies with histology and microbiological investigations. In two cases, the syndrome followed Streptococcus pneumoniae infection and in a third beta haemolytic streptococcus group A. Thus, adrenal haemorrhage alone cannot be taken to indicate meningococcal disease and other pathogens, particularly streptococcus, must be considered. PMID- 14747455 TI - Fatal Pasteurella dagmatis peritonitis and septicaemia in a patient with cirrhosis: a case report and review of the literature. AB - Pasteurella species cause zoonotic infections in humans. Human pasteurella infections usually manifest as local skin or soft tissue infection following an animal bite or scratch. Systemic infections are less common and are limited to patients at the extremes of age or those who have serious underlying disorders, including cirrhosis. Most human pasteurella infections are caused by the multocida species. We report a case of Pasteurella dagmatis peritonitis and septicaemia in a patient with cirrhosis. The infection followed a scratch inflicted by a pet dog. Despite appropriate antibiotic treatment the infection proved fatal. Spontaneous bacterial peritonitis caused by P dagmatis has not been reported previously. Pasteurella dagmatis is a relatively recently described species, which is rarely reported as a human pathogen. This species may be misidentified unless commercial identification systems are supplemented by additional biochemical tests. PMID- 14747456 TI - Normal complement C4 values do not exclude hereditary angioedema. AB - This report describes a patient with hereditary angioedema (HAE) in whom complement C4 values were consistently normal. There was a family history of HAE, for which the patient had previously been screened, but in view of her normal C4 values she was deemed unaffected. However, at 10 years of age she presented with an eight month history of episodes of swelling affecting her hands and recurrent episodes of abdominal pain over the previous few months. In view of the recent clinical history of swellings and the family history of HAE, C4 and C1 inhibitor (C1inh) were measured. The C4 concentration was found to be within the normal range but the C1inh value was low (0.07 g/litre; normal range, 0.18-0.37). The patient was started on tranexamic acid and at an outpatient review three months later her episodes of swelling were occurring less often and were less severe. Although recent papers have suggested that the diagnosis of HAE can be excluded if complement C4 concentrations are normal, this case highlights the fact that C4 concentrations can be normal in this condition, and it is recommended that both C4 and C1inh concentrations should be measured to exclude HAE. PMID- 14747457 TI - Molecular response of gastrointestinal stromal tumour after treatment with tyrosine kinase inhibitor imatinib mesylate. AB - Bleeding from the tumour site is not uncommon during the treatment of gastrointestinal stromal tumours with imatinib mesylate. It might represent an early reaction of highly vascularised tumour tissue to receptor blockade. Although often requiring emergency surgery, this is not necessarily a deleterious sign. Slow tumour regression and cystic tissue alteration may follow. Using immunohistochemistry and consecutive resection specimens, it was shown that the number of mitoses decreased significantly and MIB-1 as a marker of cell proliferation could no longer be detected. In the few tumour cells still present, the magnitude of expression of the pathognomonic marker CD117 remained unchanged. Decreases in the size of tumours responding to imatinib mesylate cannot be expected to meet the World Health Organisation or RECIST (response evaluation criteria in solid tumours) criteria. This underlines the necessity of functional imaging by positron emission tomography, contrast enhanced magnetic resonance imaging, or magnetic resonance spectroscopy to assess the response to treatment. PMID- 14747458 TI - Malignant peripheral nerve sheath tumour-like primary cutaneous malignant melanoma. AB - Malignant melanoma is known for its protean cytomorphological features, architectural patterns, and stromal changes, in addition to its ability to mimic various benign and malignant non-melanocytic tumours. Anecdotal cases of metastatic malignant melanoma simulating soft tissue sarcomas have been reported. Interestingly, this mimicry is more often seen in recurrent lesions and metastatic deposits. This report describes a case of a primary spindle cell cutaneous malignant melanoma with a prominent neural-like fascicular pattern and nuclear palisading, simulating a conventional malignant peripheral nerve sheath tumour (MPNST). Clinical, microscopic, and immunohistochemical features of the different entities included in the differential diagnosis of cutaneous spindle cell malignant tumours, such as MPNST, atypical fibroxanthoma, and spindle cell squamous cell carcinoma are discussed. Of note, the presence of an atypical epidermal or junctional component, cell pigmentation, and cell nesting, in addition to diffuse and strong reactivity for S-100 protein and other melanocytic markers, are helpful in the diagnosis of these troublesome lesions. PMID- 14747459 TI - Multifocal angiomyolipoma affecting the liver and lung without tuberous sclerosis. AB - The occurrence of angiomyolipoma (AML) in tissue other than the kidney is uncommon, as is multiple AML developing exclusively in organs other than the kidney. This report describes a case in which AML occurred multifocally in the liver and lung, but spared the kidney, and which might have been associated with tuberous sclerosis complex (TSC). A Japanese woman underwent a partial hepatectomy for a suspected malignant liver tumour at the age of 57. The tumour consisted predominantly of a trabecular arrangement of myoid cells with a sinusoidal pattern and inflammatory cell infiltration, and was diagnosed as a primary liver AML by HMB-45 immunoreactivity. Five years later, multiple nodules were found in both lungs, for which video assisted thoracic surgery was performed. The tumour showed a mixture of epithelioid cells containing HMB-45 positive material and mature lipocytes, and was subsequently diagnosed as AML. Molecular analysis of both lesions showed no allelic loss of the TSC1 and TSC2 regions. Molecular analysis of the tumours ruled out an association with TSC, and both liver and lung lesions displayed benign histological features, so that these were probably multifocal lesions of AML without TSC. PMID- 14747460 TI - A novel beta-lactamase activity from a penicillin-binding protein of Treponema pallidum and why syphilis is still treatable with penicillin. AB - Treponema pallidum, the causative agent of syphilis, is sensitive to penicillins. Yet, an abundant membrane-bound protein of this organism, Tp47, turns over penicillins. It is shown herein that the turnover process is a hydrolytic reaction that results in the corresponding penicilloates, products that have their beta-lactam bonds hydrolyzed. This is the reaction of beta-lactamases, bona fide resistance enzymes to beta-lactam antibiotics. Remarkably, the x-ray structure of Tp47 bears no resemblance to any other beta-lactamases or the related penicillin-binding proteins. Furthermore, evidence is presented that the reaction of Tp47 takes place in the absence of the zinc ion and does not involve intermediary acyl enzyme species. Hence, the beta-lactamase activity of Tp47 is the fifth known mechanism for turnover of beta-lactam antibiotics. Tp47 also exhibits a penicillin binding reaction, in the process of which the enzyme is covalently modified in the active site. The two reactions take place in two different active sites, and the events of the beta-lactamase activity are over 2,000-fold more rapid than the penicillin binding reaction. The level of beta lactamase activity is high and is held back only by a strong product-inhibition component to the catalytic process. If natural selection would result in a mutant variant of Tp47 that overcomes product inhibition for the beta-lactamase activity, a novel bona fide resistance to penicillins will emerge in Treponema, which will be a disconcerting clinical development. The physiological functions of Tp47 are not known, but it is likely that this is at least a bifunctional enzyme involved in the processing of the Treponema peptidoglycan as a substrate. PMID- 14747461 TI - Inverse regulation of rotation of F1-ATPase by the mutation at the regulatory region on the gamma subunit of chloroplast ATP synthase. AB - In F1-ATPase, the rotation of the central axis subunit gamma relative to the surrounding alpha3beta3 subunits is coupled to ATP hydrolysis. We previously reported that the introduced regulatory region of the gamma subunit of chloroplast F1-ATPase can modulate rotation of the gamma subunit of the thermophilic bacterial F1-ATPase (Bald, D., Noji, H., Yoshida, M., Hirono-Hara, Y., and Hisabori, T. (2001) J. Biol. Chem. 276, 39505-39507). The attenuated enzyme activity of this chimeric enzyme under oxidizing conditions was characterized by frequent and long pauses of rotation of gamma. In this study, we report an inverse regulation of the gamma subunit rotation in the newly engineered F1-chimeric complex whose three negatively charged residues Glu210 Asp211-Glu212 adjacent to two cysteine residues of the regulatory region derived from chloroplast F1-ATPase gamma were deleted. ATP hydrolysis activity of the mutant complex was stimulated up to 2-fold by the formation of the disulfide bond at the regulatory region by oxidation. We successfully observed inverse redox switching of rotation of gamma using this mutant complex. The complex exhibited long and frequent pauses in its gamma rotation when reduced, but the rotation rates between pauses remained unaltered. Hence, the suppression or activation of the redox-sensitive F1-ATPase can be explained in terms of the change in the rotation behavior at a single molecule level. These results obtained by the single molecule analysis of the redox regulation provide further insights into the regulation mechanism of the rotary enzyme. PMID- 14747462 TI - Concerted activation of ETS protein ER81 by p160 coactivators, the acetyltransferase p300 and the receptor tyrosine kinase HER2/Neu. AB - Activator of thyroid and retinoic acid receptor (ACTR) is overexpressed in approximately 60% of primary human breast tumors and belongs to the p160 steroid receptor coactivator family. In this study, we identified a novel interaction partner of ACTR, the ETS transcription factor ER81 that is also heavily implicated in mammary tumor formation. ACTR and related p160 family members (steroid receptor coactivator-1 and glucocorticoid receptor-interacting protein-1 (GRIP-1)) augment ER81-mediated transcription. Although ACTR and GRIP-1 can acetylate ER81, this posttranslational modification of ER81 is not required for its stimulation by ACTR or GRIP-1. In addition, ACTR collaborates with the p300 coactivator, a joint interaction partner of ACTR and ER81, to stimulate ER81 function and the ability of p300 to acetylate ER81 is indispensable for this collaboration. Furthermore, the receptor tyrosine kinase HER2/Neu, an oncoprotein particularly found overexpressed in breast tumors, cooperates with both ACTR and p300 to stimulate ER81-mediated transcription. Thus, oncogenic HER2/Neu and ACTR may synergize to orchestrate mammary tumorigenesis through the dysregulation of the transcription factor ER81 and its target genes. PMID- 14747463 TI - The ABCA1 transporter modulates late endocytic trafficking: insights from the correction of the genetic defect in Tangier disease. AB - We have previously established that the ABCA1 transporter, which plays a critical role in the lipidation of extracellular apolipoprotein acceptors, traffics between late endocytic vesicles and the cell surface (Neufeld, E. B., Remaley, A. T., Demosky, S. J., Jr., Stonik, J. A., Cooney, A. M., Comly, M., Dwyer, N. K., Zhang, M., Blanchette-Mackie, J., Santamarina-Fojo, S., and Brewer, H. B., Jr. (2001) J. Biol. Chem. 276, 27584-27590). The present study provides evidence that ABCA1 in late endocytic vesicles plays a role in cellular lipid efflux. Late endocytic trafficking was defective in Tangier disease fibroblasts that lack functional ABCA1. Consistent with a late endocytic protein trafficking defect, the hydrophobic amine U18666A retained NPC1 in abnormally tubulated, cholesterol poor, Tangier disease late endosomes, rather than cholesterol-laden lysosomes, as in wild type fibroblasts. Consistent with a lipid trafficking defect, Tangier disease late endocytic vesicles accumulated both cholesterol and sphingomyelin and were immobilized in a perinuclear localization. The excess cholesterol in Tangier disease late endocytic vesicles retained massive amounts of NPC1, which traffics lysosomal cholesterol to other cellular sites. Exogenous apoA-I abrogated the cholesterol-induced retention of NPC1 in wild type but not in Tangier disease late endosomes. Adenovirally mediated ABCA1-GFP expression in Tangier disease fibroblasts corrected the late endocytic trafficking defects and restored apoA-I-mediated cholesterol efflux. ABCA1-GFP expression in wild type fibroblasts also reduced late endosome-associated NPC1, induced a marked uptake of fluorescent apoA-I into ABCA1-GFP-containing endosomes (that shuttled between late endosomes and the cell surface), and enhanced apoA-I-mediated cholesterol efflux. The combined results of this study suggest that ABCA1 converts pools of late endocytic lipids that retain NPC1 to pools that can associate with endocytosed apoA-I, and be released from the cell as nascent high density lipoprotein. PMID- 14747464 TI - Mitochondrial deoxyribonucleotides, pool sizes, synthesis, and regulation. AB - We quantify cytosolic and mitochondrial deoxyribonucleoside triphosphates (dNTPs) from four established cell lines using a recently described method for the separation of cytosolic and mitochondrial (mt) dNTPs from as little as 10 million cells in culture (Pontarin, G., Gallinaro, L., Ferraro, P., Reichard, P., and Bianchi, V. (2003) Proc. Natl. Acad. Sci. U. S. A. 100, 12159-12164). In cycling cells the concentrations of the phosphates of thymidine, deoxycytidine, and deoxyadenosine (combining mono-, di-, and triphosphates in each case) did not differ significantly between mitochondria and cytosol, whereas deoxyguanosine phosphates were concentrated to mitochondria. We study the source and regulation of the mt dTTP pool as an example of mt dNTPs. We suggest two pathways as sources for mt dTTP: (i) import from the cytosol of thymidine diphosphate by a deoxynucleotide transporter, predominantly in cells involved in DNA replication with an active synthesis of deoxynucleotides and (ii) import of thymidine followed by phosphorylation by the mt thymidine kinase, predominantly in resting cells. Here we demonstrate that the second pathway is regulated by a mt 5' deoxyribonucleotidase (mdN). We modify the in situ activity of mdN and measure the transfer of radioactivity from [(3)H]thymidine to mt thymidine phosphates. In cycling cells lacking the cytosolic thymidine kinase, a 30-fold overproduction of mdN decreases the specific radioactivity of mt dTTP to 25%, and an 80% decrease of mdN by RNA interference increases the specific radioactivity 2-fold. These results suggest that mdN modulates the synthesis of mt dTTP by counteracting in a substrate cycle the phosphorylation of thymidine by the mt thymidine kinase. PMID- 14747465 TI - Divergence in noncognate amino acid recognition between class I and class II lysyl-tRNA synthetases. AB - Lysine insertion during coded protein synthesis requires lysyl-tRNA(Lys), which is synthesized by lysyl-tRNA synthetase (LysRS). Two unrelated forms of LysRS are known: LysRS2, which is found in eukaryotes, most bacteria, and a few archaea, and LysRS1, which is found in most archaea and a few bacteria. To compare amino acid recognition between the two forms of LysRS, the effects of l-lysine analogues on aminoacylation were investigated. Both enzymes showed stereospecificity toward the l-enantiomer of lysine and discriminated against noncognate amino acids with different R-groups (arginine, ornithine). Lysine analogues containing substitutions at other positions were generally most effective as inhibitors of LysRS2. For example, the K(i) values for aminoacylation of S-(2-aminoethyl)-l-cysteine and l-lysinamide were over 180-fold lower with LysRS2 than with LysRS1. Of the other analogues tested, only gamma aminobutyric acid showed a significantly higher K(i) for LysRS2 than LysRS1. These data indicate that the lysine-binding site is more open in LysRS2 than in LysRS1, in agreement with previous structural studies. The physiological significance of divergent amino acid recognition was reflected by the in vivo resistance to growth inhibition imparted by LysRS1 against S-(2-aminoethyl)-l cysteine and LysRS2 against gamma-aminobutyric acid. These differences in resistance to naturally occurring noncognate amino acids suggest the distribution of LysRS1 and LysRS2 contributes to quality control during protein synthesis. In addition, the specific inhibition of LysRS1 indicates it is a potential drug target. PMID- 14747466 TI - Characterization of a baculovirus enzyme with RNA ligase, polynucleotide 5' kinase, and polynucleotide 3'-phosphatase activities. AB - The end-healing and end-sealing steps of the phage T4-induced RNA restriction repair pathway are performed by two separate enzymes, a bifunctional polynucleotide 5'-kinase/3'-phosphatase and an ATP-dependent RNA ligase. Here we show that a single trifunctional baculovirus enzyme, RNA ligase 1 (Rnl1), catalyzes the identical set of RNA repair reactions. Three enzymatic activities of baculovirus Rnl1 are organized in a modular fashion within a 694-amino acid polypeptide consisting of an autonomous N-terminal RNA-specific ligase domain, Rnl1-(1-385), and a C-terminal kinase-phosphatase domain, Rnl1-(394-694). The ligase domain is itself composed of two functional units. The N-terminal module Rnl1-(1-270) contains essential nucleotidyltransferase motifs I, IV, and V and suffices for both enzyme adenylylation (step 1 of the ligation pathway) and phosphodiester bond formation at a preactivated RNA-adenylate end (step 3). The downstream module extending to residue 385 is required for ligation of a phosphorylated RNA substrate, suggesting that it is involved specifically in the second step of the end-joining pathway, the transfer of AMP from the ligase to the 5'-PO(4) end to form RNA-adenylate. The end-healing domain Rnl1-(394-694) consists of a proximal 5'-kinase module with an essential P-loop motif ((404)GSGKS(408)) and a distal 3'-phosphatase module with an essential acylphosphatase motif ((560)DLDGT(564)). Our findings have implications for the evolution of RNA repair systems and their potential roles in virus-host dynamics. PMID- 14747467 TI - Cell cycle-dependent phosphorylation of the DNA polymerase epsilon subunit, Dpb2, by the Cdc28 cyclin-dependent protein kinase. AB - DNA polymerase epsilon (Polepsilon), one of the three major eukaryotic replicative polymerases, is comprised of the essential catalytic subunit, called Pol2 in budding yeast, and three accessory subunits, only one of which, Dpb2, is essential. Polepsilon is recruited to replication origins during late G(1) phase prior to activation of replication. In this work we show that the budding yeast Dpb2 is phosphorylated in a cell cycle-dependent manner during late G(1) phase. Phosphorylation results in the appearance of a lower mobility species. The appearance of that species in vivo is dependent upon the Cdc28 cyclin-dependent protein kinase (CDK), which can directly phosphorylate Dpb2 in vitro. Either G(1) cyclin (Cln) or B-type cyclin (Clb)-associated CDK is sufficient for phosphorylation. Mapping of phosphorylation sites by mass spectrometry using a novel gel-based proteolysis protocol shows that, of the three consensus CDK phosphorylation sites, at least two, Ser-144 and Ser-616, are phosphorylated in vivo. The Cdc28 CDK phosphorylates only Ser-144 in vitro. Using site-directed mutagenesis, we show that Ser-144 is sufficient for the formation of the lower mobility form of Dpb2 in vivo. In contrast, Ser-616 appears not to be phosphorylated by Cdc28. Finally, inactivation of all three CDK consensus sites in Dpb2 results in a synthetic phenotype with the pol2-11 mutation, leading to decreased spore viability, slow growth, and increased thermosensitivity. We suggest that phosphorylation of Dpb2 during late G(1) phase at CDK consensus sites facilitates the interaction with Pol2 or the activity of Polepsilon PMID- 14747468 TI - On the roles of Saccharomyces cerevisiae Dna2p and Flap endonuclease 1 in Okazaki fragment processing. AB - Short DNA segments designated Okazaki fragments are intermediates in eukaryotic DNA replication. Each contains an initiator RNA/DNA primer (iRNA/DNA), which is converted into a 5'-flap and then removed prior to fragment joining. In one model for this process, the flap endonuclease 1 (FEN1) removes the iRNA. In the other, the single-stranded binding protein, replication protein A (RPA), coats the flap, inhibits FEN1, but stimulates cleavage by the Dna2p helicase/nuclease. RPA dissociates from the resultant short flap, allowing FEN1 cleavage. To determine the most likely process, we analyzed cleavage of short and long 5'-flaps. FEN1 cleaves 10-nucleotide fixed or equilibrating flaps in an efficient reaction, insensitive to even high levels of RPA or Dna2p. On 30-nucleotide fixed or equilibrating flaps, RPA partially inhibits FEN1. CTG flaps can form foldback structures and were inhibitory to both nucleases, however, addition of a dT(12) to the 5'-end of a CTG flap allowed Dna2p cleavage. The presence of high Dna2p activity, under reaction conditions favoring helicase activity, substantially stimulated FEN1 cleavage of tailed-foldback flaps and also 30-nucleotide unstructured flaps. Our results suggest Dna2p is not used for processing of most flaps. However, Dna2p has a role in a pathway for processing structured flaps, in which it aids FEN1 using both its nuclease and helicase activities. PMID- 14747469 TI - Inhibition of tumor invasion by genomic down-regulation of matriptase through suppression of activation of receptor-bound pro-urokinase. AB - Urokinase-type plasminogen activator (uPA) degrades the extracellular matrix and plays critical roles in tumor invasion and metastasis. Matriptase, a membrane bound serine protease, was shown to activate uPA in a uPA receptor-free, solution based study. We now investigate whether matriptase affects activation of receptor bound uPA and contributes to the invasiveness of HRA human ovarian cancer cells in vitro and tumor behavior in nude mice. Here we show the following. 1) uPA expression was effectively stimulated by TGF-beta1 in HRA cells. 2) Antisense (AS)-matriptase transfection achieved a marked inhibition of receptor-bound pro uPA activation without altering expression of uPA and uPA receptor mRNA and proteins, irrespective of whether cells were stimulated with TGF-beta1. 3) Tumor cell receptor-bound pro-uPA could be efficiently cleaved by matriptase to generate enzymatically active two-chain uPA. Thus, matriptase can substitute for plasmin in the proteolytic activation of pro-uPA to enzymatically active uPA. 4) The AS-matriptase-treated cells had a decreased ability to invade an extracellular matrix layer, as compared with control cells. 5) When the AS matriptase-treated cells were injected intraperitoneally into nude mice, the mice developed smaller tumors. Our data identify a novel role for matriptase for activation of receptor-bound uPA. PMID- 14747470 TI - Homocysteine increases the expression of vascular endothelial growth factor by a mechanism involving endoplasmic reticulum stress and transcription factor ATF4. AB - Vascular endothelial growth factor (VEGF) plays a key role in the development and progression of diabetic retinopathy. We previously demonstrated that amino acid deprivation and other inducers of endoplasmic reticulum-stress (ER stress) up regulate the expression of VEGF in the retinal-pigmented epithelial cell line ARPE-19. Because homocysteine causes ER stress, we hypothesized that VEGF expression is increased by ambient homocysteine. dl-Homocysteine-induced VEGF expression was investigated in confluent ARPE-19 cultures. Northern analysis showed that homocysteine increased steady state VEGF mRNA levels 4.4-fold. Other thiol-containing compounds, including l-homocysteine thiolactone and DTT, induced VEGF expression 7.9- and 8.8-fold. Transcriptional run-on assays and mRNA decay studies demonstrated that the increase in VEGF mRNA levels was caused by increased transcription rather than mRNA stabilization. VEGF mRNA induction paralleled that of the ER-stress gene GRP78. Homocysteine treatment caused transient phosphorylation of eIF2alpha and an increase in ATF4 protein level. Overexpression of a dominant-negative ATF4 abolished the VEGF response to homocysteine treatment and to amino acid deprivation. VEGF mRNA expression by ATF4-/- MEF did not respond to homocysteine treatment and the response was restored with expression of wild-type ATF4. These studies indicate that expression of the pro-angiogenic factor VEGF is increased by homocysteine and other thiol-containing reductive compounds via ATF4-dependent activation of VEGF transcription. PMID- 14747471 TI - Activation of the CKI-CDK-Rb-E2F pathway in full genome hepatitis C virus expressing cells. AB - Hepatitis C virus (HCV) causes persistent infection in hepatocytes, and this infection is, in turn, strongly associated with the development of hepatocellular carcinoma. To clarify the mechanisms underlying these effects, we established a Cre/loxP conditional expression system for the precisely self-trimmed HCV genome in human liver cells. Passage of hepatocytes expressing replicable full-length HCV (HCR6-Rz) RNA caused up-regulation of anchorage-independent growth after 44 days. In contrast, hepatocytes expressing HCV structural, nonstructural, or all viral proteins showed no significant changes after passage for 44 days. Only cells expressing HCR6-Rz passaged for 44 days displayed acceleration of CDK activity, hyperphosphorylation of Rb, and E2F activation. These results demonstrate that full genome HCV expression up-regulates the CDK-Rb-E2F pathway much more effectively than HCV proteins during passage. PMID- 14747472 TI - Delineation of the role of the Mre11 complex in class switch recombination. AB - Class switch recombination (CSR) is a region-specific, transcriptionally regulated, nonhomologous recombinational process that is initiated by activation induced cytidine deaminase (AID). The initial lesions in the switch (S) regions are processed and resolved, leading to a recombination of the two S regions involved. The mechanism involved in the repair and ligation of the broken DNA ends is however still unclear. Here, we describe that switching is less efficient in cells from patients with Mre11 deficiency (Ataxia-Telangiectasia-like disorder, ATLD) and, more importantly, that the switch recombination junctions resulting from the in vivo switching events are aberrant. There was a trend toward an increased usage of microhomology (> or =4 bp) at the switch junctions in both ATLD and Nijmegen breakage syndrome (NBS) patients. However, the DNA ends were not joined as "perfectly" as those from Ataxia-Telangiectasia (A-T) patients and 1-2 bp mutations or insertions were often observed. In switch junctions from ATLD patients, there were fewer base substitutions due to transitions and, most strikingly, the substitutions that occurred most often in controls, C --> T transitions, never occurred at, or close to, the junctions derived from the ATLD patients. In switch junctions from NBS patients, all base substitutions were observed at the G/C nucleotides, and transitions were preferred. These data suggest that the Mre11-Rad50-Nbs1 complex (Mre11 complex) is involved in the nonhomologous end joining pathway in CSR and that Mre11, Nbs1, and protein mutated in ataxia-telangiectasia (ATM) might have both common and independent roles in this process. PMID- 14747473 TI - Epidermal growth factor induces fibroblast contractility and motility via a protein kinase C delta-dependent pathway. AB - Myosin-based cell contractile force is considered to be a critical process in cell motility. However, for epidermal growth factor (EGF)-induced fibroblast migration, molecular links between EGF receptor (EGFR) activation and force generation have not been clarified. Herein, we demonstrate that EGF stimulation increases myosin light chain (MLC) phosphorylation, a marker for contractile force, concomitant with protein kinase C (PKC) activity in mouse fibroblasts expressing human EGFR constructs. Interestingly, PKCdelta is the most strongly phosphorylated isoform, and the preferential PKCdelta inhibitor rottlerin largely prevented EGF-induced phosphorylation of PKC substrates and MARCKS. The pathway through which EGFR activates PKCdelta is suggested by the fact that the MEK-1 inhibitor U0126 and the phosphatidylinositol 3-kinase inhibitor LY294002 had no effect on PKCdelta activation, whereas lack of PLCgamma signaling resulted in delayed PKCdelta activation. EGF-enhanced MLC phosphorylation was prevented by a specific MLC kinase inhibitor ML-7 and the PKC inhibitors chelerythrine chloride and rottlerin. Further indicating that PKCdelta is required, a dominant-negative PKCdelta construct or RNAi-mediated PKCdelta depletion also prevented MLC phosphorylation. In the absence of PLC signaling, MLC phosphorylation and cell force generation were delayed similarly to PKCdelta activation. All of the interventions that blocked PKCdelta activation or MLC phosphorylation abrogated EGF-induced cell contractile force generation and motility. Our results suggest that PKCdelta activation is responsible for a major part of EGF-induced fibroblast contractile force generation. Hence, we identify here a new pathway helping to govern cell motility, with PLC signaling playing a role in activation of PKCdelta to promote the acute phase of EGF-induced MLC activation. PMID- 14747474 TI - Dimerization and processing of procaspase-9 by redox stress in mitochondria. AB - We studied the mechanism of intra-mitochondrial death initiator caspase-9 activation by a redox response, in which hydrogen peroxide (H(2)O(2)) caused a subtle decrease in the inner membrane potential (Deltapsim) with little evidence of cytochrome c release. Initiation of the intra-mitochondrial autocleavage of procaspase-9 preceded the onset of caspase cascade induction in the cytosol. Purified mitochondria demonstrated procaspase-9 processing and releasing abilities when exposed to H(2)O(2). Bcl-2 overexpression caused accumulation of the active form caspase-9 in the mitochondria, rendering the cells resistant to the redox stress. Intriguingly, disulfide-bonded dimers of autoprocessed caspase 9 were generated in the mitochondria in the pre-apoptotic phase. Using a substrate-analog inhibitor, dimer formation of procaspase-9 was also detectable inside the mitochondria. Furthermore, thiol reductant thioredoxin blocked the caspase-9 activation step and the cell death induction. Thus, redox stress responsive thiol-disulfide converting reactions in the mitochondrion seemed to mediate procaspase-9 assembly that allows autoprocessing. This study offers an explanation for the recent observation that Apaf-1-null cells can execute apoptosis, which can be blocked by Bcl-2, and supports the proposition that the cytochrome c-Apaf-1-procaspase-9 complex functions in the caspase amplification rather than in its initiation. PMID- 14747475 TI - Molecular characterization of covalent complexes between tissue transglutaminase and gliadin peptides. AB - Tissue transglutaminase (TG2) modifies proteins and peptides by transamidation or deamidation of specific glutamine residues. TG2 also has a central role in the pathogenesis of celiac disease. The enzyme is both the target of disease-specific autoantibodies and generates deamidated gliadin peptides recognized by intestinal T cells from patients. Incubation of TG2 with gliadin peptides also results in the formation of covalent TG2-peptide complexes. Here we report the characterization of complexes between TG2 and two immunodominant gliadin peptides. Two types of covalent complexes were found; the peptides are either linked via a thioester bond to the active site cysteine of TG2 or via isopeptide bonds to particular lysine residues of the enzyme. We quantified the number of gliadin peptides bound to TG2 under different conditions. After 30 min of incubation of TG2 at 1 microm with an equimolar ratio of peptides to TG2, approximately equal amounts of peptides were bound by thioester and isopeptide linkage. At higher peptide to TG2 ratios, more than one peptide was linked to TG2, and isopeptide bond formation dominated. The lysine residues in TG2 that act as acyl acceptors were identified by matrix assisted laser desorption ionization and nanoelectrospray mass spectrometry and tandem mass spectrometry analysis of proteolytic digests of the TG2-peptide complexes. At a high molar excess of gliadin peptides to TG2 altogether six lysine residues of TG2 were found to participate in isopeptide bond formation. The results are relevant to the understanding of how antibodies to TG2 are formed in celiac disease. PMID- 14747476 TI - AML1/RUNX1 increases during G1 to S cell cycle progression independent of cytokine-dependent phosphorylation and induces cyclin D3 gene expression. AB - AML1/RUNX1, a member of the core binding factor (CBF) family stimulates myelopoiesis and lymphopoiesis by activating lineage-specific genes. In addition, AML1 induces S phase entry in 32Dcl3 myeloid or Ba/F3 lymphoid cells via transactivation. We now found that AML1 levels are regulated during the cell cycle. 32Dcl3 and Ba/F3 cell cycle fractions were prepared using elutriation. Western blotting and a gel shift/supershift assay demonstrated that endogenous CBF DNA binding and AML1 levels were increased 2-4-fold in S and G(2)/M phase cells compared with G(1) cells. In addition, G(1) arrest induced by mimosine reduced AML1 protein levels. In contrast, AML1 RNA did not vary during cell cycle progression relative to actin RNA. Analysis of exogenous Myc-AML1 or AML1-ER demonstrated a significant reduction in G(1) phase cells, whereas levels of exogenous DNA binding domain alone were constant, lending support to the conclusion that regulation of AML1 protein stability contributes to cell cycle variation in endogenous AML1. However, cytokine-dependent AML1 phosphorylation was independent of cell cycle phase, and an AML1 mutant lacking two ERK phosphorylation sites was still cell cycle-regulated. Inhibition of AML1 activity with the CBFbeta-SMMHC or AML1-ETO oncoproteins reduced cyclin D3 RNA expression, and AML1 bound and activated the cyclin D3 promoter. Signals stimulating G(1) to S cell cycle progression or entry into the cell cycle in immature hematopoietic cells might do so in part by inducing AML1 expression, and mutations altering pathways regulating variation in AML1 stability potentially contribute to leukemic transformation. PMID- 14747477 TI - Alpha-Pal/NRF-1 regulates the promoter of the human integrin-associated protein/CD47 gene. AB - Integrin-associated protein (IAP or CD47) is expressed in a variety of tissues, including the nervous system and immune system. To understand how cells control the expression of the IAP gene, we cloned the 5'-proximal region of the human IAP gene and investigated IAP promoter activity by transient transfection. RT-PCR confirmed the expression of IAP transcripts in human neuroblastoma IMR-32 and hepatoma HepG2 cells. Deletion analysis identified a core promoter of the human IAP gene located between nucleotide positions -232 and -12 relative to the translation initiation codon in these two cell lines. Site-directed mutagenesis and gel electrophoretic mobility shift assay identified a alpha-Pal/NRF-1 binding element within the IAP core promoter. Supershift assays using the alpha-Pal/NRF-1 antiserum confirmed the binding of this transcription factor on the alpha-Pal/NRF 1 site. Overexpression of the DNA binding domain of alpha-Pal/NRF-1 in cells enhanced DNA-alpha-Pal/NRF-1 binding in vitro. Furthermore, overexpression of full-length alpha-Pal/NRF-1 significantly enhanced IAP promoter activity while overexpression of dominant-negative mutant reduced promoter activity both in the cultured human cell lines and primary mouse cortical cells. These results revealed that alpha-Pal/NRF-1 is an essential transcription factor in the regulation of human IAP gene expression. PMID- 14747478 TI - The human Frizzled 6 (HFz6) acts as a negative regulator of the canonical Wnt. beta-catenin signaling cascade. AB - Previously we have cloned the human Frizzled 1 (HFz1) and shown that it transmits the Wnt-3a-induced canonical pathway. We also cloned the human Frizzled 6 (HFz6) and show in the present study that, as opposed to HFz1, HFz6 did not activate the canonical Wnt pathway following exposure to various Wnts, whether belonging to the Wnt-1 or to the Wnt-5a group. Moreover we show that HFz6 repressed Wnt-3a induced canonical signaling when co-expressed with HFz1. HFz6 repressed the canonical Wnt cascade activated also by various Wnt signaling intracellular mediators such as Dishevelled-1, a stabilized beta-catenin(S33Y) mutant, and LiCl mediated repression of glycogen synthase kinase-3beta activity. Removal of HFz6 N'- or C'-terminal sequences abolished HFz6 repressive activity. As the HFz6 repressive effect was not associated with a decrease in the level of beta catenin, it is suggested that HFz6 does not affect beta-catenin stabilization, implying that HFz6 transmits a repressive signaling that cross-talks with and inhibits the canonical Wnt pathway downstream of beta-catenin destruction complex. HFz6 did not affect the level of nuclear T-cell factor 4 (TCF4) nor did it affect beta-catenin.TCF4 complex formation. However, electrophoretic mobility shift assays indicated that HFz6 repressed the binding of TCF/lymphoid enhancer factor transcription factors to target DNA. Moreover we present data suggesting that HFz6 activates the transforming growth factor-beta-activated kinase-NEMO like kinase pathway that blocks TCF/lymphoid enhancer factor binding to target promoters, thereby inhibiting the ability of beta-catenin to activate transcription of Wnt target genes. PMID- 14747479 TI - Standards set for CT calcium screening but its clinical value remains unclear. PMID- 14747480 TI - Pediatric drug studies required by law. PMID- 14747481 TI - Knee replacement underused, says panel: useful option when nonsurgical therapies fail. PMID- 14747491 TI - Quality of care and satisfaction among patients isolated for infection control. PMID- 14747492 TI - Quality of care and satisfaction among patients isolated for infection control. PMID- 14747493 TI - Quality of care and satisfaction among patients isolated for infection control. PMID- 14747494 TI - Assessing exposure to toxic gases in Bhopal. PMID- 14747495 TI - Ozone and asthma. PMID- 14747496 TI - Genetic vs hormonal factors in lipid metabolism in women. PMID- 14747497 TI - Effect of poverty on emotional symptoms in children. PMID- 14747498 TI - Realizing the benefits of practical clinical trials. PMID- 14747499 TI - Migraine as a risk factor for subclinical brain lesions. AB - CONTEXT: Clinical series have suggested an increased prevalence of cerebral infarction and white matter lesions (WMLs) in migraine patients. It is not known whether these lesions are prevalent in the general migraine population. OBJECTIVES: To compare the prevalence of brain infarcts and WMLs in migraine cases and controls from the general population and to identify migraine characteristics associated with these lesions. DESIGN: Cross-sectional, prevalence study of population-based sample of Dutch adults aged 30 to 60 years. PARTICIPANTS: Randomly selected patients with migraine with aura (n = 161), patients with migraine without aura (n = 134), and controls (n = 140), who were frequency matched to cases for age, sex, and place of residence. Nearly 50% of the cases had not been previously diagnosed by a physician. MAIN OUTCOME MEASURES: Brain magnetic resonance images were evaluated for infarcts, by location and vascular supply territory, and for periventricular WMLs (PVWMLs) and deep WMLs (DWMLs). The odds ratios (ORs) and 95% confidence intervals (CIs) of these brain lesions compared with controls were examined by migraine subtype (with or without aura) and monthly attack frequency (<1 attack, > or =1 attack), controlling for cardiovascular risk factors and use of vasoconstrictor migraine agents. All participants underwent a standard neurological examination. RESULTS: No participants reported a history of stroke or transient ischemic attack or had relevant abnormalities at standard neurological examination. We found no significant difference between patients with migraine and controls in overall infarct prevalence (8.1% vs 5.0%). However, in the cerebellar region of the posterior circulation territory, patients with migraine had a higher prevalence of infarct than controls (5.4% vs 0.7%; P =.02; adjusted OR, 7.1; 95% CI, 0.9 55). The adjusted OR for posterior infarct varied by migraine subtype and attack frequency. The adjusted OR was 13.7 (95% CI, 1.7-112) for patients with migraine with aura compared with controls. In patients with migraine with a frequency of attacks of 1 or more per month, the adjusted OR was 9.3 (95% CI, 1.1-76). The highest risk was in patients with migraine with aura with 1 attack or more per month (OR, 15.8; 95% CI, 1.8-140). Among women, the risk for high DWML load (top 20th percentile of the distribution of DWML load vs lower 80th percentile) was increased in patients with migraine compared with controls (OR, 2.1; 95% CI, 1.0 4.1); this risk increased with attack frequency (highest in those with > or =1 attack per month: OR, 2.6; 95% CI, 1.2-5.7) but was similar in patients with migraine with or without aura. In men, controls and patients with migraine did not differ in the prevalence of DWMLs. There was no association between severity of PVWMLs and migraine, irrespective of sex or migraine frequency or subtype. CONCLUSIONS: These population-based findings suggest that some patients with migraine with and without aura are at increased risk for subclinical lesions in certain brain areas. PMID- 14747500 TI - Prognostic value of placental growth factor in patients with acute chest pain. AB - CONTEXT: Experimental data suggest that placental growth factor (PlGF), a member of the vascular endothelial growth factor family, acts as a primary inflammatory instigator of atherosclerotic plaque instability and thus may be useful as a risk predicting biomarker in patients with acute coronary syndromes (ACS). OBJECTIVE: To determine whether blood levels of PlGF predict risk for death or nonfatal myocardial infarction in patients with acute chest pain. DESIGN, SETTING, AND PATIENTS: Measurement of PlGF levels as well as levels of markers of myocardial necrosis (troponin T [TnT]), platelet activation (soluble CD40 ligand [sCD40L]), and inflammation (high-sensitivity C-reactive protein [hsCRP]) in an inception cohort of 547 patients with angiographically validated ACS participating in the CAPTURE (c7E3 Fab Anti-Platelet Therapy in Unstable Refractory Angina) trial and in a heterogeneous cohort of 626 patients presenting with acute chest pain to an emergency department in Germany between December 1996 and March 1999. MAIN OUTCOME MEASURE: Risk for death or nonfatal myocardial infarction after 30 days. RESULTS: In patients with ACS, elevated PlGF levels (>27.0 ng/L; 40.8% of patients) indicated a markedly increased risk of events at 30 days (14.8% vs 4.9%; unadjusted hazard ratio [HR], 3.34; 95% confidence interval [CI], 1.79 6.24; P<.001). In a multivariable model, elevated levels of TnT (HR, 1.83; 95% CI, 1.05-3.86; P =.03), sCD40L (HR, 2.65; 95% CI, 1.41-4.99; P =.002), and PlGF (HR, 3.03; 95% CI, 1.54-5.95; P<.001) were independent predictors, while elevated hsCRP level was not (HR, 0.98; 95% CI, 0.53-1.98; P =.94). In patients with acute chest pain, elevated levels of PlGF predicted risk (21.2% vs 5.3%) (unadjusted: HR, 4.80; 95% CI, 2.81-8.21; P<.001; adjusted: HR, 3.00; 95% CI, 1.68-5.38; P<.001). Patients negative for all 3 markers (TnT, sCD40L, and PlGF) were at very low cardiac risk (7 days: no event; 30 days: 2.1% event rate). CONCLUSIONS: Plasma PlGF levels may be an independent biomarker of adverse outcome in patients with suspected ACS. A single initial measurement of plasma PlGF appears to extend the predictive and prognostic information gained from traditional inflammatory markers. PMID- 14747501 TI - Effect of breast augmentation on the accuracy of mammography and cancer characteristics. AB - CONTEXT: Breast augmentation is not associated with an increased risk of breast cancer; however, implants may interfere with the detection of breast cancer thereby delaying cancer diagnosis in women with augmentation. OBJECTIVE: To determine whether mammography accuracy and tumor characteristics are different for women with and without augmentation. DESIGN, SETTING, AND PARTICIPANTS: A prospective cohort of 137 women with augmentation and 685 women without augmentation diagnosed with breast cancer between January 1, 1995, and October 15, 2002, matched (1:5) by age, race/ethnicity, previous mammography screening, and mammography registry, and 10 533 women with augmentation and 974 915 women without augmentation and without breast cancer among 7 mammography registries in Denver, Colo; Lebanon, NH; Albuquerque, NM; Chapel Hill, NC; San Francisco, Calif; Seattle, Wash; and Burlington, Vt. MAIN OUTCOME MEASURES: Comparison between women with and without augmentation of mammography performance measures and cancer characteristics, including invasive carcinoma or ductal carcinoma in situ, tumor stage, nodal status, size, grade, and estrogen-receptor status. RESULTS: Among asymptomatic women, the sensitivity of screening mammography based on the final assessment was lower in women with breast augmentation vs women without (45.0% [95% confidence interval [CI], 29.3%-61.5%] vs 66.8% [95% CI, 60.4%-72.8%]; P =.008), and specificity was slightly higher in women with augmentation (97.7% [95% CI, 97.4%-98.0%] vs 96.7% [95% CI, 96.6%-96.7%]; P<.001). Among symptomatic women, both sensitivity and specificity were lower for women with augmentation compared with women without but these differences were not significant. Tumors were of similar stage, size, estrogen-receptor status, and nodal status but tended to be lower grade (P =.052) for women with breast augmentation vs without. CONCLUSIONS: Breast augmentation decreases the sensitivity of screening mammography among asymptomatic women but does not increase the false-positive rate. Despite the lower accuracy of mammography in women with augmentation, the prognostic characteristics of tumors are not influenced by augmentation. PMID- 14747502 TI - Association between cholesterol level and mortality in dialysis patients: role of inflammation and malnutrition. AB - CONTEXT: Total cholesterol level is inversely associated with mortality in dialysis patients, a group at high risk of cardiovascular disease (CVD). This paradox may be explained by systemic inflammation and/or malnutrition, which are associated with lower cholesterol levels and higher mortality. OBJECTIVE: To determine the relationship between cholesterol level and outcome in patients undergoing dialysis, accounting for inflammation and malnutrition. DESIGN, SETTING, AND PARTICIPANTS: Prospective study of 823 patients enrolled from October 1995 to June 1998 who recently initiated dialysis, from 79 clinics, classified by absence or presence of inflammation and/or malnutrition (defined as serum albumin levels <3.6 mg/dL, C-reactive protein > or =10 mg/L, or interleukin 6 > or =3.09 pg/mL). MAIN OUTCOME MEASURES: All-cause and cardiovascular disease mortality. RESULTS: During a median follow-up of 2.4 years, 324 deaths (159 CVD deaths), 153 renal transplantations, and 10 losses to follow-up occurred. Average serum cholesterol level was lower in the presence of inflammation/malnutrition than in its absence. In a Cox model adjusted for age, race, and sex, a 40-mg/dL (1.0-mmol/L) increment in baseline total serum cholesterol level was associated with a decreased risk of all-cause mortality overall (relative hazard [RH], 0.92; 95% confidence interval [CI], 0.87-0.98) and in the presence of inflammation/malnutrition (RH, 0.89; CI, 0.84-0.95). In contrast, serum cholesterol level was associated with an increased risk in the absence of inflammation/malnutrition (RH, 1.32; 95% CI, 1.07-1.63). For CVD mortality, an inverse trend was not statistically significant in the presence of inflammation/malnutrition, and a positive association was evident in the absence of inflammation/malnutrition (RH, 1.41; 95% CI, 1.04-1.89). Further adjustment for traditional CVD risk factors, dialysis modality, comorbidity, and inflammatory markers attenuated the inverse association but strengthened the positive association. CONCLUSIONS: The inverse association of total cholesterol level with mortality in dialysis patients is likely due to the cholesterol lowering effect of systemic inflammation and malnutrition, not to a protective effect of high cholesterol concentrations. These findings support treatment of hypercholesterolemia in this population. PMID- 14747503 TI - Penetrance of the fragile X-associated tremor/ataxia syndrome in a premutation carrier population. AB - CONTEXT: Premutation expansions (55-200 CGG repeats) of the fragile X mental retardation 1 (FMR1) gene are frequent in the general population, with estimated prevalences of 1 per 259 females and 1 per 813 males. Several articles have recently described the presence of late-onset neurological symptoms in male carriers of premutation (FMR1) alleles. The main clinical features described in this newly identified syndrome are cerebellar ataxia and intention tremor. Additional documented symptoms include short-term memory loss, executive functional deficits, cognitive decline, parkinsonism, peripheral neuropathy, lower-limb proximal muscle weakness, and autonomic dysfunction. OBJECTIVE: To study the penetrance of the fragile X-associated tremor/ataxia syndrome (FXTAS) among premutation carriers. DESIGN, SETTING, AND PARTICIPANTS: Family-based study of 192 individuals (premutation carriers and controls) whose families belong to the Northern or Southern California Fragile X Associations. Data were collected (March 2002-April 2003) through a survey and a standardized neurological examination, which was videotaped and subsequently scored in a blinded fashion. MAIN OUTCOME MEASURES: Penetrance of intention tremor and ataxia among adult carriers (aged > or =50 years) of premutation expansions of the FMR1 gene. RESULTS: Data from the survey of 192 individuals demonstrated an age-related penetrance of the combination of reported intention tremor and gait ataxia in male carriers (17%, 38%, 47%, and 75% [lower-bound estimates] for participants aged 50-59, 60-69, 70-79, and > or =80 years, respectively). The male carrier group had an age-adjusted 13-fold increased risk (95% confidence interval, 3.9 25.4; P =.003) of combined intention tremor and gait ataxia when compared with male controls. The clinical examination data from 93 individuals demonstrated that male carriers experienced more difficulties on each of 3 standardized neurological rating scales compared with controls (P<.05). Female carrier scores were also higher than those of female controls (P<.05) on 2 of the 3 neurological rating scales, but no participant was identified with probable or definite FXTAS. CONCLUSIONS: The study demonstrates that older male carriers of premutation alleles of the FMR1 gene are at high risk of developing FXTAS. Since male premutation carriers are relatively common in the general population, older men with ataxia and intention tremor should be screened for the FMR1 mutation, especially if these signs are accompanied by parkinsonism, autonomic dysfunction, or cognitive decline, regardless of family history. PMID- 14747504 TI - Communication of randomization in childhood leukemia trials. AB - CONTEXT: Most children diagnosed as having leukemia become research subjects in randomized clinical trials (RCTs), but little is known about how randomization is explained to or understood by parents. OBJECTIVE: To investigate physicians' explanation and parental understanding of randomization in childhood leukemia RCTs. DESIGN AND SETTING: A multisite study of the informed consent communication process for RCTs of childhood leukemia. Consecutive cases were recruited from pediatric oncology inpatient wards at 6 US children's hospitals associated with major academic medical centers from July 1, 1999, until December 31, 2001. The informed consent conferences were observed and audiotaped, and the information obtained was coded and analyzed. Parents were interviewed shortly after the conference to ascertain their understanding. PARTICIPANTS: Parents and members of the health care team who participated in 137 informed consent conferences for children with newly diagnosed acute leukemia. MAIN OUTCOME MEASURES: Observed explanations of randomization and parental understanding of randomization after the consent conference. RESULTS: Randomization was explained by physicians in 83% of cases and a consent document was presented during the conference in 95% of cases. Interviews after the conference demonstrated that 68 (50%) of 137 parents did not understand randomization. Parents of racial minority and lower socioeconomic status were less likely to understand randomization (P<.001 for each). Discussion of specific clinical trial details and the presence of a nurse during the conference were associated with understanding. Eighty-four percent of children were enrolled in a leukemia trial. CONCLUSIONS: Despite oral and written explanation, half of the parents in this study did not understand randomization for childhood leukemia trials. To make informed consent more effective, future research must seek to improve communication during this critical interchange. PMID- 14747505 TI - How do institutional review boards apply the federal risk and benefit standards for pediatric research? AB - CONTEXT: Federal regulations allow children in the United States to be enrolled in clinical research only when the institutional review board (IRB) determines that the risks are minimal or a minor increase over minimal, or that the research offers a prospect of direct benefit. Despite this reliance on IRBs, no data exist on how IRBs apply the risk and benefit categories for pediatric research. OBJECTIVE: To determine how IRB chairpersons apply the federal risk and benefit categories for pediatric research. DESIGN, SETTING, AND PARTICIPANTS: Telephone survey, conducted between May and August 2002 of 188 randomly selected chairpersons of IRBs in the Unites States. The survey consisted of 21 questions to assess the application of federal risk standards to research procedures, whether certain interventions offer a prospect of direct benefit to participating children, and the extent to which IRBs use the federal definition of minimal risk when categorizing the risks of research procedures in children. MAIN OUTCOME MEASURES: Responses regarding categorization of the risk level and direct benefits of pediatric research procedures. RESULTS: A single blood draw was the only procedure categorized as minimal risk by a majority (152 or 81%) of the 188 respondents. An electromyogram was categorized as minimal or a minor increase over minimal risk by 100 (53%) and as more than a minor increase over minimal risk by 77 (41%). Allergy skin testing was categorized as minimal risk by 43 IRB chairpersons (23%), a minor increase over minimal risk by 81 (43%), and more than a minor increase over minimal risk by 51 (27%). Regarding benefits, 113 chairpersons (60%) considered added psychological counseling to be a direct benefit, while participant payment was considered a direct benefit by 10% (n = 19). CONCLUSIONS: Application of the federal risk and benefit categories for pediatric research by IRB chairpersons is variable and sometimes contradicted by the available data on risks and the regulations themselves. To protect children from excessive risks while allowing appropriate research, IRB chairpersons need guidance on applying the federal risk and benefit categories and also need data on the risks children face in daily life and during routine physical or psychological tests. PMID- 14747506 TI - Supporting family caregivers at the end of life: "they don't know what they don't know". AB - Even for patients receiving complex, intensive medical care for serious and life threatening illness, family caregiving is typically at the core of what sustains patients at the end of life. The amorphous relationship between physicians and the families of patients at the end of life presents both challenges and opportunities for which physicians may be unprepared. Families play important roles in the practical and emotional aspects of patient care and in decision making at the end of life. At the same time, family members may carry significant burdens as a result of their work. Through the perspectives of the wife, daughter, and home care nurse of a patient who died from pancreatic cancer, we illustrate the range of family caregiver experiences and suggest potentially helpful physician interventions. We describe 5 burdens of family caregiving (time and logistics, physical tasks, financial costs, emotional burdens and mental health risks, and physical health risks) and review the responsibilities of physicians to family caregivers. Based on available evidence, we identify 5 areas of opportunity for physicians to be of service to family members caring for patients at the end of life, including promoting excellent communication with family, encouraging appropriate advance care planning and decision making, supporting home care, demonstrating empathy for family emotions and relationships, and attending to family grief and bereavement. In caring well for family caregivers at the end of life, physicians may not only improve the experiences of patients and family but also find greater sustenance and meaning in their own work. PMID- 14747507 TI - Overcoming the false dichotomy of "curative" vs "palliative" care for late-stage HIV/AIDS: "let me live the way I want to live, until I can't". PMID- 14747508 TI - Is migraine a progressive brain disease? PMID- 14747509 TI - Determining the appropriateness of including children in clinical research: how thick is the ice? PMID- 14747510 TI - JAMA patient page. Childhood leukemia. PMID- 14747511 TI - Acetylcholine, histamine, and cognition: two sides of the same coin. PMID- 14747513 TI - Changes in acetylcholine extracellular levels during cognitive processes. AB - Measuring the changes in neurotransmitter extracellular levels in discrete brain areas is considered a tool for identifying the neuronal systems involved in specific behavioral responses or cognitive processes. Acetylcholine (ACh) is the first neurotransmitter whose diffusion from the central nervous system was investigated and whose extracellular levels variations were correlated to changes in neuronal activity. This was done initially by means of the cup technique and then by the microdialysis technique. The latter, notwithstanding some technical limitations, makes it possible to detect variations in extracellular levels of ACh in unrestrained, behaving animals. This review summarizes and discusses the results obtained investigating the changes in ACh release during performance of operant tasks, exposition to novel stimuli, locomotor activity, and the performance of spatial memory tasks, working memory, and place preference memory tasks. Activation of the forebrain cholinergic system has been demonstrated in many tasks and conditions in which the environment requires the animal to analyze novel stimuli that may represent a threat or offer a reward. The sustained cholinergic activation, demonstrated by high levels of extracellular ACh observed during the behavioral paradigms, indicates that many behaviors occur within or require the facilitation provided by the cholinergic system to the operation of pertinent neuronal pathways. PMID- 14747515 TI - The role of muscarinic and nicotinic cholinergic neurotransmission in aversive conditioning: comparing pavlovian fear conditioning and inhibitory avoidance. PMID- 14747512 TI - Septohippocampal acetylcholine: involved in but not necessary for learning and memory? AB - The neurotransmitter acetylcholine (ACh) has been accorded an important role in supporting learning and memory processes in the hippocampus. Cholinergic activity in the hippocampus is correlated with memory, and restoration of ACh in the hippocampus after disruption of the septohippocampal pathway is sufficient to rescue memory. However, selective ablation of cholinergic septohippocampal projections is largely without effect on hippocampal-dependent learning and memory processes. We consider the evidence underlying each of these statements, and the contradictions they pose for understanding the functional role of hippocampal ACh in memory. We suggest that although hippocampal ACh is involved in memory in the intact brain, it is not necessary for many aspects of hippocampal memory function. PMID- 14747514 TI - Acetylcholine and olfactory perceptual learning. AB - Olfactory perceptual learning is a relatively long-term, learned increase in perceptual acuity, and has been described in both humans and animals. Data from recent electrophysiological studies have indicated that olfactory perceptual learning may be correlated with changes in odorant receptive fields of neurons in the olfactory bulb and piriform cortex. These changes include enhanced representation of the molecular features of familiar odors by mitral cells in the olfactory bulb, and synthetic coding of multiple coincident odorant features into odor objects by cortical neurons. In this paper, data are reviewed that show the critical role of acetylcholine (Ach) in olfactory system function and plasticity, and cholinergic modulation of olfactory perceptual learning at both the behavioral and cortical level. PMID- 14747516 TI - The cholinergic lesion of Alzheimer's disease: pivotal factor or side show? AB - A profound loss of cortical cholinergic innervation is a nearly invariant feature of advanced Alzheimer's disease (AD). The temporal course of this lesion and its relationship to other aspects of the disease have not yet been fully clarified. Despite assertions to the contrary, a review of the evidence suggests that a perturbation of cholinergic innervation is likely to be present even in the very early stages of AD. This cholinergic lesion is unlikely to be a major determinant of the clinical symptoms or of the neuropathological lesions. Nonetheless, it almost certainly contributes to the severity of the cognitive and behavioral deficits, especially in the areas of memory and attention. The cholinergic lesion may also influence the progression of the neuropathological process through complex interactions with amyloidogenesis, tau phosphorylation and neuroplasticity. PMID- 14747517 TI - Nicotinic acetylcholine receptors in sensory cortex. AB - Acetylcholine release in sensory neocortex contributes to higher-order sensory function, in part by activating nicotinic acetylcholine receptors (nAChRs). Molecular studies have revealed a bewildering array of nAChR subtypes and cellular actions; however, there is some consensus emerging about the major nAChR subtypes and their functions in sensory cortex. This review first describes the systems-level effects of activating nAChRs in visual, somatosensory, and auditory cortex, and then describes, as far as possible, the underlying cellular and synaptic mechanisms. A related goal is to examine if sensory cortex can be considered a model system for cortex in general, because the use of sensory stimuli to activate neural circuits physiologically is helpful for understanding mechanisms of systems-level function and plasticity. A final goal is to highlight the emerging role of nAChRs in developing sensory cortex, and the adverse impact of early nicotine exposure on subsequent sensory-cognitive function. PMID- 14747518 TI - Nicotinic cholinergic synaptic mechanisms in the ventral tegmental area contribute to nicotine addiction. AB - Tobacco use is a major health problem that is estimated to cause 4 million deaths a year worldwide. Nicotine is the main addictive component of tobacco. It acts as an agonist to activate and desensitize nicotinic acetylcholine receptors (nAChRs). A component of nicotine's addictive power is attributable to actions on the mesolimbic dopaminergic system, which serves a fundamental role in the acquisition of behaviors that are inappropriately reinforced by addictive drugs. Here we show that nicotine, in the same concentration and time ranges as obtained from tobacco, has three main actions that regulate the activity of midbrain dopamine (DA) neurons. Nicotine first activates and then desensitizes nAChRs on the DA neurons. This process directly excites the DA neurons for a short period of time before the nAChRs desensitize. Nicotine also enhances glutamatergic excitation and decreases GABAergic inhibition onto DA neurons. These events increase the probability for synaptic plasticity, such as long-term potentiation. The short-lived direct excitation of the DA neurons coupled with the enhanced glutamatergic afferent activity provides the presynaptic and postsynaptic coincidence necessary to initiate synaptic potentiation. In total, these synaptic events lead to a relatively long-lasting heightened activity of midbrain DA neurons. Consistent with other summarized studies, this work indicates that the synaptic changes normally associated with learning and memory can be influenced and commandeered during the nicotine addiction process. PMID- 14747519 TI - Dynamic changes in acetylcholine output in the medial striatum during place reversal learning. AB - The present studies explored the role of the medial striatum in learning when task contingencies change. Experiment 1 examined whether the medial striatum is involved in place reversal learning. Testing occurred in a modified cross-maze across two consecutive sessions. Injections of the local anesthetic, bupivacaine, into the medial striatum, did not impair place acquisition, but impaired place reversal learning. The reversal-learning deficit was due to an inability to maintain the new choice pattern following the initial shift. Experiment 2 determined whether changes in acetylcholine (ACh) output occur during the acquisition or reversal learning of a place discrimination. Extracellular ACh output from the medial striatum was assessed in samples collected at 6-min intervals using in vivo microdialysis during behavioral testing. ACh output did not change from basal levels during place acquisition. During reversal learning, ACh output significantly increased as rats began to learn the new choice pattern, and returned to near basal levels as a rat reliably executed the new place strategy. The present results suggest that the medial striatum may be critical for flexible adaptations involving spatial information, and that ACh actions in this area enable the shifting of choice patterns when environmental conditions change. PMID- 14747520 TI - Cholinergic modulation of visual attention and working memory: dissociable effects of basal forebrain 192-IgG-saporin lesions and intraprefrontal infusions of scopolamine. AB - Two experiments examined the effects of reductions in cortical cholinergic function on performance of a novel task that allowed for the simultaneous assessment of attention to a visual stimulus and memory for that stimulus over a variable delay within the same test session. In the first experiment, infusions of the muscarinic receptor antagonist scopolamine into the medial prefrontal cortex (mPFC) produced many omissions but did not impair rats' ability to correctly detect a brief visual stimulus. However, these animals were highly impaired in remembering the location of that stimulus following a delay period, although in a delay-independent manner. In the second experiment, another group of animals with selective 192 IgG-saporin lesions of the nucleus basalis magnocellularis (nBM) were not impaired under conditions of low-attentional demand. However, when the stimulus duration was reduced, a significant memory impairment was observed, but similar to the results of the first experiment, the nBM-lesioned animals were not impaired in attentional accuracy, although aspects of attention were compromised (e.g., omissions). These findings demonstrate that (1) cortical cholinergic depletion produces dissociable deficits in attention and memory, depending on the task demands, (2) delay-independent mnemonic deficits produced by scopolamine are probably due to impairments other than simple inattention, and (3) working memory deficits are not simply dependent on attentional difficulties per se. Together, these findings implicate the nBM cortical cholinergic system in both attentional and mnemonic processing. PMID- 14747521 TI - Extensive lesions of cholinergic basal forebrain neurons do not impair spatial working memory. AB - A recent study suggests that lesions to all major areas of the cholinergic basal forebrain in the rat (medial septum, horizontal limb of the diagonal band of Broca, and nucleus basalis magnocellularis) impair a spatial working memory task. However, this experiment used a surgical technique that may have damaged cerebellar Purkinje cells. The present study tested rats with highly selective lesions of cholinergic neurons in all major areas of the basal forebrain on a spatial working memory task in the radial arm maze. In postoperative testing, there were no significant differences between lesion and control groups in working memory, even with a delay period of 8 h, with the exception of a transient impairment during the first 2 d of postoperative testing at shorter delays (0 or 2 h). This finding corroborates other results that indicate that the cholinergic basal forebrain does not play a significant role in spatial working memory. Furthermore, it underscores the presence of intact memory functions after cholinergic basal forebrain damage, despite attentional impairments that follow these lesions, demonstrated in other task paradigms. PMID- 14747522 TI - Perirhinal cortex muscarinic receptor blockade impairs taste recognition memory formation. AB - The relevance of perirhinal cortical cholinergic and glutamatergic neurotransmission for taste recognition memory and learned taste aversion was assessed by microinfusions of muscarinic (scopolamine), NMDA (AP-5), and AMPA (NBQX) receptor antagonists. Infusions of scopolamine, but not AP5 or NBQX, prevented the consolidation of taste recognition memory using attenuation of neophobia as an index. In addition, learned taste aversion in both short- and long-term memory tests was exclusively impaired by scopolamine. These data provide neurochemical support for the theory that cholinergic activity of the perirhinal cortex participates in the formation of the taste memory trace and that it is independent of the NMDA and AMPA receptor activity. These results support the idea that cholinergic neurotransmission in the perirhinal cortex is also essential for acquisition and consolidation of taste recognition memory. PMID- 14747523 TI - Cholinergic modulation of the hippocampus during encoding and retrieval of tone/shock-induced fear conditioning. AB - We investigated the role of acetylcholine (ACh) during encoding and retrieval of tone/shock-induced fear conditioning with the aim of testing Hasselmo's cholinergic modulation model of encoding and retrieval using a task sensitive to hippocampal disruption. Lesions of the hippocampus impair acquisition and retention of contextual conditioning with no effect on tone conditioning. Cholinergic antagonists also impair acquisition of contextual conditioning. Saline, scopolamine, or physostigmine was administered directly into the CA3 subregion of the hippocampus 10 min before rats were trained on a tone/shock induced fear conditioning paradigm. Freezing behavior was used as the measure of learning. The scopolamine group froze significantly less during acquisition to the context relative to controls. The scopolamine group also froze less to the context test administered 24 h posttraining. A finer analysis of the data revealed that scopolamine disrupted encoding but not retrieval. The physostigmine group initially froze less during acquisition to the context, although this was not significantly different from controls. During the context test, the physostigmine group froze less initially but quickly matched the freezing levels of controls. A finer analysis of the data indicated that physostigmine disrupted retrieval but not encoding. These results suggest that increased ACh levels are necessary for encoding new spatial contexts, whereas decreased ACh levels are necessary for retrieving previously learned spatial contexts. PMID- 14747525 TI - Galantamine facilitates acquisition of a trace-conditioned eyeblink response in healthy, young rabbits. AB - Previous work has demonstrated that drugs increasing brain concentrations of acetylcholine can enhance cognition in aging and brain-damaged organisms. The present study assessed whether galantamine (GAL), an allosteric modulator of nicotinic cholinergic receptors and weak acetylcholinesterase inhibitor, could improve acquisition and retention of an eyeblink (EB) classical conditioning task in healthy, young animals. We trained 24 rabbits (n = 8/group) in a 1000-msec trace Pavlovian EB conditioning paradigm in which a tone conditioned stimulus (CS) was presented for 500 msec, followed by a 500-msec trace period in which no stimuli were presented. A 100-msec corneal airpuff was the unconditioned stimulus (US). Acquisition sessions, consisting of 100 trials each, occurred daily for 10 consecutive days, followed by 3 d of extinction training. Animals were treated with one of three doses of GAL (0.0-3.0 mg/kg) prior to each session. Animals that received 3.0 mg/kg GAL showed significantly more EB conditioned responses (CRs) in fewer training trials than animals receiving either 1.5 mg/kg GAL or vehicle injections. GAL had no effect on CR performance during extinction. Pseudoconditioning control experiments, consisting of 200 explicitly unpaired tone-puff presentations indicated that GAL did not increase reactivity to the CS or US. These findings indicate that GAL may improve acquisition of moderately difficult associative learning tasks in healthy young organisms. PMID- 14747524 TI - Galantamine facilitates acquisition of hippocampus-dependent trace eyeblink conditioning in aged rabbits. AB - Cholinergic systems are critical to the neural mechanisms mediating learning. Reduced nicotinic cholinergic receptor (nAChR) binding is a hallmark of normal aging. These reductions are markedly more severe in some dementias, such as Alzheimer's disease. Pharmacological central nervous system therapies are a means to ameliorate the cognitive deficits associated with normal aging and aging related dementias. Trace eyeblink conditioning (EBC), a hippocampus- and forebrain-dependent learning paradigm, is impaired in both aged rabbits and aged humans, attributable in part to cholinergic dysfunction. In the present study, we examined the effects of galantamine (3 mg/kg), a cholinesterase inhibitor and nAChR allosteric potentiating ligand, on the acquisition of trace EBC in aged (30 33 months) and young (2-3 months) female rabbits. Trace EBC involves the association of a conditioned stimulus (CS) with an unconditioned stimulus (US), separated by a stimulus-free trace interval. Repeated CS-US pairings results in the development of the conditioned eyeblink response (CR) prior to US onset. Aged rabbits receiving daily injections of galantamine (Aged/Gal) exhibited significant improvements compared with age-matched controls in trials to eight CRs in 10 trial block criterion (P = 0.0402) as well as performance across 20 d of training [F(1,21) = 5.114, P = 0.0345]. Mean onset and peak latency of CRs exhibited by Aged/Gal rabbits also differed significantly [F(1,21) = 6.120/6.582, P = 0.0220/0.0180, respectively] compared with age-matched controls, resembling more closely CR timing of young drug and control rabbits. Galantamine did not improve acquisition rates in young rabbits compared with age-matched controls. These data indicate that by enhancing nicotinic and muscarinic transmission, galantamine is effective in offsetting the learning deficits associated with decreased cholinergic transmission in the aging brain. PMID- 14747526 TI - Recruitment of replication protein A by the papillomavirus E1 protein and modulation by single-stranded DNA. AB - With the exception of viral proteins E1 and E2, papillomaviruses depend heavily on host replication machinery for replication of their viral genome. E1 and E2 are known to recruit many of the necessary cellular replication factors to the viral origin of replication. Previously, we reported a physical interaction between E1 and the major human single-stranded DNA (ssDNA)-binding protein, replication protein A (RPA). E1 was determined to bind to the 70-kDa subunit of RPA, RPA70. In this study, using E1-affinity coprecipitation and enzyme-linked immunosorbent assay-based interaction assays, we show that E1 interacts with the major ssDNA-binding domain of RPA. Consistent with our previous report, no measurable interaction between E1 and the two smaller subunits of RPA was detected. The interaction of E1 with RPA was substantially inhibited by ssDNA. The extent of this inhibition was dependent on the length of the DNA. A 31 nucleotide (nt) oligonucleotide strongly inhibited the E1-RPA interaction, while a 16-nt oligonucleotide showed an intermediate level of inhibition. In contrast, a 10-nt oligonucleotide showed no observable effect on the E1-RPA interaction. This inhibition was not dependent on the sequence of the DNA. Furthermore, ssDNA also inhibited the interaction of RPA with papillomavirus E2, simian virus 40 T antigen, human polymerase alpha-primase, and p53. Taken together, our results suggest a potential role for ssDNA in modulating RPA-protein interactions, in particular, the RPA-E1 interactions during papillomavirus DNA replication. A model for recruitment of RPA by E1 during papillomavirus DNA replication is proposed. PMID- 14747527 TI - Fusion to C3d enhances the immunogenicity of the E2 glycoprotein of type 2 bovine viral diarrhea virus. AB - The use of DNA and protein subunit vaccines in animals provides an opportunity to introduce vaccines that are arguably the safest that can be developed. For that reason, considerable effort is under way to devise methods of enhancing the immunogenicity of such vaccines. Seven years ago it was shown that fusing complement fragment C3d to hen egg lysozyme (HEL) enhanced the immunogenicity of HEL 10,000-fold. Based on this observation, we decided to evaluate the effect of C3d on the immunogenicity of the E2 protein of bovine viral diarrhea virus (BVDV). E2 is the major target of neutralizing antibody during BVDV infection. To test the effect of C3d on E2 immunogenicity, expression cassettes encoding a secreted form of E2 alone (E2s) or E2 fused to three copies of murine C3d (E2s C3d) were constructed. The proteins were purified from the supernatants of transfected cells and used to immunize mice. The immune response was monitored by an enzyme-linked immunosorbent assay (ELISA) for E2s-specific antibody and by a virus neutralization test. The ELISA results indicated that the E2s-C3d protein is 10,000-fold more immunogenic than the E2s protein alone. The maximum primary immune response was elicited with <0.1 microg of E2s-C3d protein without an adjuvant. In addition, we have shown for the first time that high levels of anti E2s and neutralizing antibodies can be elicited when this same low concentration of E2s-C3d is used to both prime and boost the immune response. We conclude that the E2s-C3d fusion protein has significant potential as a subunit vaccine against BVDV infection. PMID- 14747528 TI - Dramatic effects of 2-bromo-5,6-dichloro-1-beta-D-ribofuranosyl benzimidazole riboside on the genome structure, packaging, and egress of guinea pig cytomegalovirus. AB - The halogenated benzimidazoles BDCRB (2-bromo-5,6-dichloro-1-beta-D riborfuranosyl benzimidazole riboside) and TCRB (2,5,6-trichloro-1-beta-D riborfuranosyl benzimidazole riboside) were the first compounds shown to inhibit cleavage and packaging of herpesvirus genomes. Both inhibit the formation of unit length human cytomegalovirus (HCMV) genomes by a poorly understood mechanism (M. R. Underwood et al., J. Virol. 72:717-715, 1998; P. M. Krosky et al., J. Virol. 72:4721-4728, 1998). Because the simple genome structure of guinea pig cytomegalovirus (GPCMV) provides a useful model for the study of herpesvirus DNA packaging, we investigated the effects of BDCRB on GPCMV. GPCMV proved to be sensitive to BDCRB (50% inhibitory concentration = 4.7 microM), although somewhat less so than HCMV. In striking contrast to HCMV, however, a dose of BDCRB sufficient to reduce GPCMV titers by 3 logs (50 microM) had no effect on the quantity of GPCMV genomic DNA that was formed in infected cells. Electron microscopy revealed that this DNA was in fact packaged within intranuclear capsids, but these capsids failed to egress from the nucleus and failed to protect the DNA from nuclease digestion. The terminal structure of genomes formed in the presence of BDCRB was also altered. Genomes with ends lacking a terminal repeat at the right end, which normally exist in an equimolar ratio with those having one copy of the repeat at the right end, were selectively eliminated by BDCRB treatment. At the left end, BDCRB treatment appeared to induce heterogeneous truncations such that 2.7 to 4.9 kb of left-end-terminal sequences were missing. These findings suggest that BDCRB induces premature cleavage events that result in truncated genomes packaged within capsids that are permeable to nuclease. Based on these and other observations, we propose a model for duplication of herpesvirus terminal repeats during the cleavage and packaging process that is similar to one proposed for bacteriophage T7 (Y. B. Chung, C. Nardone, and D. C. Hinkle, J. Mol. Biol. 216:939-948, 1990). PMID- 14747529 TI - Active Akt and functional p53 modulate apoptosis in Abelson virus-transformed pre B cells. AB - Suppression of apoptosis is an important feature of the Abelson murine leukemia virus (Ab-MLV) transformation process. During multistep transformation, Ab-MLV infected pre-B cells undergo p53-dependent apoptosis during the crisis phase of transformation. Even once cells are fully transformed, an active v-Abl protein tyrosine kinase is required to suppress apoptosis because cells transformed by temperature-sensitive (ts) kinase mutants undergo rapid apoptosis after a shift to the nonpermissive temperature. However, inactivation of the v-Abl protein by a temperature shift interrupts signals transmitted via multiple pathways, making it difficult to identify those that are critically important for the suppression of apoptosis. To begin to dissect these pathways, we tested the ability of an SH2 domain Ab-MLV mutant, P120/R273K, to rescue aspects of the ts phenotype of pre-B cells transformed by the conditional kinase domain mutant. The P120/R273K mutant suppressed apoptosis at the nonpermissive temperature, a phenotype correlated with its ability to activate Akt. Apoptosis also was suppressed at the nonpermissive temperature by constitutively active Akt and in p53-null pre-B cells transformed with the ts kinase domain mutant. These data indicate that an intact Src homology 2 (SH2) domain is not critical for apoptosis suppression and suggest that signals transmitted through Akt and p53 play an important role in the response. PMID- 14747530 TI - Conservation of a stepwise, energy-sensitive pathway involving HP68 for assembly of primate lentivirus capsids in cells. AB - Previously we have described a stepwise, energy-dependent pathway for human immunodeficiency virus type 1 (HIV-1) capsid assembly in a cell-free system. In this pathway, Gag polypeptides utilize the cellular factor HP68 and assemble into immature capsids by way of assembly intermediates that have defined biochemical characteristics. Here we address whether this pathway is universally conserved among primate lentiviruses and can be observed in mammalian cells. We demonstrate that HIV-2 Gag associates with human HP68 in a cell-free system and that Gag proteins of HIV-2, simian immunodeficiency virus SIVmac239, and SIVagm associate with endogenous HP68 in primate cells, as is seen for HIV-1. Analysis of primate cells expressing lentivirus Gag proteins revealed Gag-containing complexes with the same sedimentation values as seen for previously described HIV-1 assembly intermediates in the cell-free system (10S, 80-150S, and 500S). These complexes fit criteria for assembly intermediates as judged by energy sensitivity, pattern of HP68 association, and the failure of specific complexes to be formed by assembly-incompetent Gag mutants. We also demonstrate that virus-like particles released from cells do not appear to contain HP68, suggesting that HP68 is released from Gag upon completion of capsid assembly in cells, as was observed previously in the cell-free system. Together these findings support a model in which all primate lentivirus capsids assemble by a conserved pathway of HP68 containing, energy-dependent assembly intermediates that have specific biochemical features. PMID- 14747531 TI - High physiological levels of LMP1 result in phosphorylation of eIF2 alpha in Epstein-Barr virus-infected cells. AB - LMP1 is an Epstein-Barr virus (EBV)-encoded membrane protein essential for the proliferation of EBV-infected lymphoblasts (E. Kilger, A. Kieser, M. Baumann, and W. Hammerschmidt, EMBO J. 17:1700-1709, 1998). LMP1 also inhibits gene expression and induces cytostasis in transfected cells when it is expressed at levels as little as twofold higher than the average for EBV-positive lymphoblasts (M. Sandberg, A. Kaykas, and B. Sugden, J. Virol. 74:9755-9761, 2000; A. Kaykas and B. Sugden, Oncogene 19:1400-1410, 2000). We have found that in three different clones of EBV-infected lymphoblasts the levels of expression of LMP1 in individual cells in each clone ranged over 100-fold. This difference is due to a difference in levels of the LMP1 transcript. In these clones, cells expressing high levels of LMP1 incorporated less BrdU. We also found that induction of expression of LMP1 or of a derivative of LMP1 with its transmembrane domain fused to green fluorescent protein instead of its carboxy-terminal signaling domain resulted in phosphorylation of eIF2 alpha in EBV-negative Burkitt's lymphoma cells. This induction of phosphorylation of eIF2 alpha was also detected in EBV infected lymphoblasts, in which high levels of LMP1 correlated with high levels of phosphorylation of eIF2 alpha. Our results indicate that inhibition of gene expression and of cell proliferation by LMP1 occurs normally in EBV-infected cells. PMID- 14747532 TI - Selective induction of Th2-attracting chemokines CCL17 and CCL22 in human B cells by latent membrane protein 1 of Epstein-Barr virus. AB - Chemokines are likely to play important roles in the pathophysiology of diseases associated with Epstein-Barr virus (EBV). Here, we have analyzed the repertoire of chemokines expressed by EBV-infected B cells. EBV infection of B cells induced expression of TARC/CCL17 and MDC/CCL22, which are known to attract Th2 cells and regulatory T cells via CCR4, and also upregulated constitutive expression of MIP 1 alpha/CCL3, MIP-1 beta/CCL4, and RANTES/CCL5, which are known to attract Th1 cells and cytotoxic T cells via CCR5. Accordingly, EBV-immortalized B cells secreted these chemokines, especially CCL3, CCL4, and CCL22, in large quantities. EBV infection or stable expression of LMP1 also induced CCL17 and CCL22 in a B cell line, BJAB. The inhibitors of the TRAF/NF-kappa B pathway (BAY11-7082) and the p38/ATF2 pathway (SB202190) selectively suppressed the expression of CCL17 and CCL22 in EBV-immortalized B cells and BJAB-LMP1. Consistently, transient transfection assays using CCL22 promoter-reporter constructs demonstrated that two NF-kappa B sites and a single AP-1 site were involved in the activation of the CCL22 promoter by LMP1. Finally, serum CCL22 levels were significantly elevated in infectious mononucleosis. Collectively, LMP1 induces CCL17 and CCL22 in EBV-infected B cells via activation of NF-kappa B and probably ATF2. Production of CCL17 and CCL22, which attract Th2 and regulatory T cells, may help EBV-infected B cells evade immune surveillance by Th1 cells. However, the concomitant production of CCL3, CCL4, and CCL5 by EBV-infected B cells may eventually attract Th1 cells and cytotoxic T cells, leading to elimination of EBV infected B cells at latency III and to selection of those with limited expression of latent genes. PMID- 14747533 TI - The herpes simplex virus ICP0 RING finger domain inhibits IRF3- and IRF7-mediated activation of interferon-stimulated genes. AB - Virus infection induces a rapid cellular response in cells characterized by the induction of interferon. While interferon itself does not induce an antiviral response, it activates a number of interferon-stimulated genes that collectively function to inhibit virus replication and spread. Previously, we and others reported that herpes simplex virus type 1 (HSV-1) induces an interferon independent antiviral response in the absence of virus replication. Here, we report that the HSV-1 proteins ICP0 and vhs function in concert to disable the host antiviral response. In particular, we show that ICP0 blocks interferon regulatory factor IRF3- and IRF7-mediated activation of interferon-stimulated genes and that the RING finger domain of ICP0 is essential for this activity. Furthermore, we demonstrate that HSV-1 modifies the IRF3 pathway in a manner different from that of the small RNA viruses most commonly studied. PMID- 14747534 TI - Human immunodeficiency virus type 1 Nef associates with lipid rafts to downmodulate cell surface CD4 and class I major histocompatibility complex expression and to increase viral infectivity. AB - Lipid rafts are membrane microdomains that are functionally distinct from other membrane regions. We have shown that 10% of human immunodeficiency virus type 1 (HIV-1) Nef expressed in SupT1 cells is present in lipid rafts and that this represents virtually all of the membrane-associated Nef. To determine whether raft targeting, rather than simply membrane localization, has functional significance, we created a Nef fusion protein (LAT-Nef) containing the N-terminal 35 amino acids from LAT, a protein that is exclusively localized to rafts. Greater than 90% of the LAT-Nef protein was found in the raft fraction. In contrast, a mutated form, lacking two cysteine palmitoylation sites, showed less than 5% raft localization. Both proteins were equally expressed and targeted nearly exclusively to membranes. The LAT-Nef protein was more efficient than its nonraft mutant counterpart at downmodulating both cell surface CD4 and class I major histocompatibility complex (MHC) expression, as well as in enhancing first round infectivity and being incorporated into virus particles. This demonstrates that targeting of Nef to lipid rafts is mechanistically important for all of these functions. Compared to wild-type Nef, LAT-Nef downmodulated class I MHC nearly as effectively as the wild-type Nef protein, but was only about 60% as effective for CD4 downmodulation and 30% as effective for infectivity enhancement. Since the LAT-Nef protein was found entirely in rafts while the wild type Nef protein was distributed 10% in rafts and 90% in the soluble fraction, our results suggest that class I MHC downmodulation by Nef may be performed exclusively by raft-bound Nef. In contrast, CD4 downmodulation and infectivity enhancement may require a non-membrane-bound Nef component as well as the membrane-bound form. PMID- 14747535 TI - Latent membrane protein 2A inhibits transforming growth factor-beta 1-induced apoptosis through the phosphatidylinositol 3-kinase/Akt pathway. AB - Latent membrane protein 2A (LMP2A) blocks B-cell receptor signal transduction in vitro by binding the Syk and Lyn protein tyrosine kinases. As well as blocking B cell signal transduction, LMP2A has been shown to activate the phosphatidylinositol 3-kinase (PI3-K)/Akt pathway, which acts as a survival signal in both B cells and epithelial cells. Transforming growth factor beta 1 (TGF-beta 1) is a multifunctional cytokine that plays important roles in regulating cell growth and differentiation in many biological systems. The loss of the growth-inhibitory response to the TGF-beta 1 signal is found in many cancers and is widely thought to promote tumor development. In this study, we found that LMP2A induced the phosphorylation of Akt (serine 473) in Burkitt's lymphoma cell line Ramos and in gastric carcinoma cell line HSC-39 and partially enhanced cell viability following TGF-beta 1 treatment. In addition, LMP2A partially inhibited TGF-beta 1-induced DNA fragmentation and cleavage of poly(ADP ribose) polymerase (PARP). In the presence of LY294002, an inhibitor of PI3-K, the LMP2A-mediated inhibitory effects on TGF-beta 1-induced DNA fragmentation and cleavage of PARP were alleviated. Furthermore, LMP2A did not alter the levels of expression of type I and type II TGF-beta 1 receptors. Taken together, these results suggest that LMP2A may inhibit TGF-beta 1-mediated apoptosis through activation of the PI3-K/Akt pathway. PMID- 14747536 TI - Innate cellular response to virus particle entry requires IRF3 but not virus replication. AB - Mammalian cells respond to virus infections by eliciting both innate and adaptive immune responses. One of the most effective innate antiviral responses is the production of alpha/beta interferon and the subsequent induction of interferon stimulated genes (ISGs), whose products collectively limit virus replication and spread. Following viral infection, interferon is produced in a biphasic fashion that involves a number of transcription factors, including the interferon regulatory factors (IRFs) 1, 3, 7, and 9. In addition, virus infection has been shown to directly induce ISGs in the absence of prior interferon production through the activation of IRF3. This process is believed to require virus replication and results in IRF3 hyperphosphorylation, nuclear localization, and proteasome-mediated degradation. Previously, we and others demonstrated that herpes simplex virus type 1 (HSV-1) induces ISGs and an antiviral response in fibroblasts in the absence of both interferon production and virus replication. In this report, we show that the entry of enveloped virus particles from diverse virus families elicits a similar innate response. This process requires IRF3, but not IRF1, IRF7, or IRF9. Following virus replication, the large DNA viruses HSV-1 and vaccinia virus effectively inhibit ISG mRNA accumulation, whereas the small RNA viruses Newcastle disease virus, Sendai virus, and vesicular stomatitis virus do not. In addition, we found that IRF3 hyperphosphorylation and degradation do not correlate with ISG and antiviral state induction but instead serve as a hallmark of productive virus replication, particularly following a high multiplicity infection. Collectively, these data suggest that virus entry triggers an innate antiviral response mediated by IRF3 and that subsequent virus replication results in posttranslational modification of IRF3, such as hyperphosphorylation, depending on the nature of the incoming virus. PMID- 14747537 TI - Novel single-cell-level phenotypic assay for residual drug susceptibility and reduced replication capacity of drug-resistant human immunodeficiency virus type 1. AB - Human immunodeficiency virus type 1 (HIV-1)-infected individuals who develop drug resistant virus during antiretroviral therapy may derive benefit from continued treatment for two reasons. First, drug-resistant viruses can retain partial susceptibility to the drug combination. Second, therapy selects for drug resistant viruses that may have reduced replication capacities relative to archived, drug-sensitive viruses. We developed a novel single-cell-level phenotypic assay that allows these two effects to be distinguished and compared quantitatively. Patient-derived gag-pol sequences were cloned into an HIV-1 reporter virus that expresses an endoplasmic reticulum-retained Env-green fluorescent protein fusion. Flow cytometric analysis of single-round infections allowed a quantitative analysis of viral replication over a 4-log dynamic range. The assay faithfully reproduced known in vivo drug interactions occurring at the level of target cells. Simultaneous analysis of single-round infections by wild type and resistant viruses in the presence and absence of the relevant drug combination divided the benefit of continued nonsuppressive treatment into two additive components, residual virus susceptibility to the drug combination and selection for drug-resistant variants with diminished replication capacities. In some patients with drug resistance, the dominant circulating viruses retained significant susceptibility to the combination. However, in other cases, the dominant drug-resistant viruses showed no residual susceptibility to the combination but had a reduced replication capacity relative to the wild-type virus. In this case, simplification of the regimen might still allow adequate suppression of the wild-type virus. In a third pattern, the resistant viruses had no residual susceptibility to the relevant drug regimen but nevertheless had a replication capacity equivalent to that of wild-type virus. In such cases, there is no benefit to continued treatment. Thus, the ability to simultaneously analyze residual susceptibility and reduced replication capacity of drug-resistant viruses may provide a basis for rational therapeutic decisions in the setting of treatment failure. PMID- 14747538 TI - Low doses of hepadnavirus induce infection of the lymphatic system that does not engage the liver. AB - Woodchuck hepatitis virus (WHV), which is closely related to human hepatitis B virus and is considered to be principally hepatotropic, invades the host's lymphatic system and persists in lymphoid cells independently of whether the infection is symptomatic and serologically evident or concealed. In this study, we show, with the woodchuck model of hepatitis B, that hepadnavirus can establish an infection that engages the lymphatic system, but not the liver, and persists in the absence of virus serological markers, including antiviral antibodies. This primary occult infection is caused by wild-type virus invading the host at a quantity usually not greater than 10(3) virions. It is characterized by trace virus replication progressing in lymphatic organs and peripheral lymphoid cells that, with time, may also spread to the liver. The infection is transmissible to virus-naive hosts as an asymptomatic, indefinitely long, occult carriage of small amounts of biologically competent virus. In contrast to residual silent WHV persistence, which normally endures after the resolution of viral hepatitis and involves the liver, primary occult infection restricted to the lymphatic system does not protect against reinfection with a large, liver-pathogenic WHV dose; however, the occult infection is associated with a swift recovery from hepatitis caused by the superinfection. Our study documents that the lymphatic system is the primary target of WHV infection when small quantities of virions invade a susceptible host. PMID- 14747539 TI - Perforin and gamma interferon-mediated control of coronavirus central nervous system infection by CD8 T cells in the absence of CD4 T cells. AB - Infection of the central nervous system (CNS) with the neurotropic JHM strain of mouse hepatitis virus produces acute and chronic demyelination. The contributions of perforin-mediated cytolysis and gamma interferon (IFN-gamma) secretion by CD8(+) T cells to the control of infection and the induction of demyelination were examined by adoptive transfer into infected SCID recipients. Untreated SCID mice exhibited uncontrolled virus replication in all CNS cell types but had little or no demyelination. Memory CD8(+) T cells from syngeneic wild-type (wt), perforin-deficient, or IFN-gamma-deficient (GKO) donors all trafficked into the infected CNS in the absence of CD4(+) T cells and localized to similar areas. Although CD8(+) T cells from all three donors suppressed virus replication in the CNS, GKO CD8(+) T cells expressed the least antiviral activity. A distinct viral antigen distribution in specific CNS cell types revealed different mechanisms of viral control. While wt CD8(+) T cells inhibited virus replication in all CNS cell types, cytolytic activity in the absence of IFN-gamma suppressed the infection of astrocytes, but not oligodendroglia. In contrast, cells that secreted IFN-gamma but lacked cytolytic activity inhibited replication in oligodendroglia, but not astrocytes. Demyelination was most severe following viral control by wt CD8(+) T cells but was independent of macrophage infiltration. These data demonstrate the effective control of virus replication by CD8(+) T cells in the absence of CD4(+) T cells and support the necessity for the expression of distinct effector mechanisms in the control of viral replication in distinct CNS glial cell types. PMID- 14747540 TI - In vivo expansion of the residual tumor antigen-specific CD8+ T lymphocytes that survive negative selection in simian virus 40 T-antigen-transgenic mice. AB - Mice that express the viral oncoprotein simian virus 40 (SV40) large T antigen (T Ag) as a transgene provide useful models for the assessment of the state of the host immune response in the face of spontaneous tumor progression. Line SV11 (H2(b)) mice develop rapidly progressing choroid plexus tumors due to expression of full-length T-Ag from the SV40 promoter. In addition, T-Ag expression in the thymus of SV11 mice results in the deletion of CD8(+) T cells specific for the three H2(b)-restricted immunodominant epitopes of T-Ag. Whether CD8(+) T cells specific for the immunorecessive H2-D(b)-restricted epitope V of T-Ag survive negative selection in SV11 mice has not been determined. Immunization of SV11 mice with rVV-ES-V, a recombinant vaccinia virus expressing epitope V as a minigene, resulted in the induction of weak, but reproducible, epitope V-specific cytotoxic T-lymphocyte (CTL) responses. This weak lytic response corresponded with a decreased frequency of epitope V-specific CTL that could be recruited in SV11 mice. In addition, CTL lines derived from rVV-ES-V-immunized SV11 mice had reduced avidities compared to that seen with CTL derived from healthy mice. Despite this initial weak response, significant numbers of epitope V-specific CD8(+) T cells were detected in SV11 mice ex vivo following a priming-boosting approach and these cells demonstrated high avidity for epitope V. The results suggest that low numbers of tumor-reactive CD8(+) T cells with high avidity for epitope V survive negative selection in SV11 mice but can be expanded by specific boosting approaches in the tumor bearing host. PMID- 14747541 TI - Phenotype of a herpes simplex virus type 1 mutant that fails to express immediate early regulatory protein ICP0. AB - Herpes simplex virus type 1 (HSV-1) immediate-early (IE) regulatory protein ICP0 is required for efficient progression of infected cells into productive lytic infection, especially in low-multiplicity infections of limited-passage human fibroblasts. We have used single-cell-based assays that allow detailed analysis of the ICP0-null phenotype in low-multiplicity infections of restrictive cell types. The major conclusions are as follows: (i) there is a threshold input multiplicity above which the mutant virus replicates normally; (ii) individual cells infected below the threshold multiplicity have a high probability of establishing a nonproductive infection; (iii) such nonproductively infected cells have a high probability of expressing IE products at 6 h postinfection; (iv) even at 24 h postinfection, IE protein-positive nonproductively infected human fibroblast cells exceed the number of cells that lead to plaque formation by up to 2 orders of magnitude; (v) expression of individual IE proteins in a proportion of the nonproductively infected cells is incompletely coordinated; (vi) the nonproductive cells can also express early gene products at low frequencies and in a stochastic manner; and (vii) significant numbers of human fibroblast cells infected at low multiplicity by an ICP0-deficient virus are lost through cell death. We propose that in the absence of ICP0 expression, HSV-1 infected human fibroblasts can undergo a great variety of fates, including quiescence, stalled infection at a variety of different stages, cell death, and, for a minor population, initiation of formation of a plaque. PMID- 14747542 TI - Identification of a common HLA-DP4-restricted T-cell epitope in the conserved region of the respiratory syncytial virus G protein. AB - The cellular immune response to respiratory syncytial virus (RSV) is important in both protection and immunopathogenesis. In contrast to HLA class I, HLA class II restricted RSV-specific T-cell epitopes have not been identified. Here, we describe the generation and characterization of two human RSV-specific CD4(+)-T cell clones (TCCs) associated with type 0-like cytokine profiles. TCC 1 was specific for the matrix protein and restricted over HLA-DPB1*1601, while TCC 2 was specific for the attachment protein G and restricted over either HLA DPB1*0401 or -0402. Interestingly, the latter epitope is conserved in both RSV type A and B viruses. Given the high allele frequencies of HLA-DPB1*0401 and 0402 worldwide, this epitope could be widely recognized and boosted by recurrent RSV infections. Indeed, peptide stimulation of peripheral blood mononuclear cells from healthy adults resulted in the detection of specific responses in 8 of 13 donors. Additional G-specific TCCs were generated from three of these cultures, which recognized the identical (n = 2) or almost identical (n = 1) HLA-DP4 restricted epitope as TCC 2. No significant differences were found between the capacities of cell lines obtained from infants with severe (n = 41) or mild (n = 46) RSV lower respiratory tract infections to function as antigen-presenting cells to the G-specific TCCs, suggesting that the severity of RSV disease is not linked to the allelic frequency of HLA-DP4. In conclusion, we have identified an RSV G-specific human T helper cell epitope restricted by the widely expressed HLA class II alleles DPB1*0401 and -0402. Its putative role in protection and/or immunopathogenesis remains to be determined. PMID- 14747543 TI - Activation of the early-late switch in adenovirus type 5 major late transcription unit expression by L4 gene products. AB - The adenovirus major late transcription unit (MLTU) encodes multiple proteins from five regions, L1 to L5, through differential splicing and polyadenylation. MLTU expression is temporally regulated; only a single product from L1 (52/55K) is expressed prior to replication, but a subsequent switch, the mechanism of which has not been defined, leads to full expression that encompasses L1 IIIa and all L2 to L5 products. Transfection of a plasmid containing the complete MLTU gave a full array of proteins in proportions similar to those in a late infection, and in a time course, the temporal pattern of expression in a natural infection was reproduced. However, a plasmid truncated after the L3 poly(A) site exclusively expressed the L1 52/55K protein and was defective in the switch to full gene expression from L1 to L3. The L4 33K protein, supplied in trans, was sufficient to upregulate cytoplasmic mRNA for MLTU products characteristic of the late pattern of expression to levels comparable to those produced by the full length MLTU. There was a corresponding increase in expression of the L1 IIIa, L2, and L3 proteins, except hexon. Hexon protein expression additionally required both the L4 100K protein in trans and sequences downstream of the L3 poly(A) site in cis. These results indicate that induction of L4 protein expression is a key event in the early-late switch in MLTU expression, which we propose is precipitated by small amounts of L4 expression in a feed-forward activation mechanism. PMID- 14747544 TI - CD46 is a cellular receptor for bovine viral diarrhea virus. AB - Various monoclonal antibodies (MAbs) that recognize cell surface proteins on bovine cells were previously shown to efficiently block infection with bovine viral diarrhea virus (BVDV) (C. Schelp, I. Greiser-Wilke, G. Wolf, M. Beer, V. Moennig, and B. Liess, Arch. Virol. 140:1997-2009, 1995). With one of these MAbs, a 50- to 58-kDa protein was purified from calf thymus by immunoaffinity chromatography. Microchemical analysis of two internal peptides revealed significant sequence homology to porcine and human CD46. The cDNA of bovine CD46 (CD46(bov)) was cloned and further characterized. Heterologously expressed CD46(bov) was detected by the MAb used for purification. A putative function of CD46(bov) as a BVDV receptor was studied with respect to virus binding and susceptibility of nonpermissive cells. While the expression of CD46(bov) correlated well with the binding of [(3)H]uridine-labeled BVDV, the susceptibility of cells nonpermissive for BVDV was not observed. However, the expression of CD46(bov) resulted in a significant increase in the susceptibility of porcine cells to BVDV. These results provide strong evidence that CD46(bov) serves as a cellular receptor for BVDV. PMID- 14747545 TI - Nuclear factor kappa B-dependent activation of the antiapoptotic bfl-1 gene by the Epstein-Barr virus latent membrane protein 1 and activated CD40 receptor. AB - Suppression of the cellular apoptotic program by the oncogenic herpesvirus Epstein-Barr virus (EBV) is central to both the establishment of latent infection and the development of EBV-associated malignancies. We have previously shown that expression of the EBV latent membrane protein 1 (LMP1) in Burkitt's lymphoma cell lines leads to increased mRNA levels from the cellular antiapoptotic bfl-1 gene (also known as A1). Furthermore, ectopic expression of Bfl-1 in an EBV-positive cell line exhibiting a latency type 1 infection protects against apoptosis induced by growth factor deprivation (B. N. D'Souza, M. Rowe, and D. Walls, J. Virol. 74:6652-6658, 2000). We now report that LMP1 drives bfl-1 promoter activity through interactions with components of the tumor necrosis factor receptor (TNFR)/CD40 signaling pathway. We present evidence that this process is NF-kappa B dependent, involves the recruitment of TNFR-associated factor 2, and is mediated to a greater extent by the carboxyl-terminal activating region 2 (CTAR2) relative to the CTAR1 domain of LMP1. Activation of CD40 receptor also led to increased bfl-1 mRNA levels and an NF-kappa B-dependent increase in bfl-1 promoter activity in Burkitt's lymphoma-derived cell lines. We have delineated a 95-bp region of the promoter that functions as an LMP1-dependent transcriptional enhancer in this cellular context. This sequence contains a novel NF-kappa B-like binding motif that is essential for transactivation of bfl-1 by LMP1, CD40, and the NF-kappa B subunit protein p65. These findings highlight the role of LMP1 as a mediator of EBV-host cell interactions and may indicate an important route by which it exerts its cellular growth transforming properties. PMID- 14747547 TI - A cytomegalovirus inhibitor of gamma interferon signaling controls immunoproteasome induction. AB - Both human and mouse cytomegaloviruses (HCMV and MCMV) avoid peptide presentation through the major histocompatibility complex (MHC) class I pathway to CD8(+) T cells. Within the MHC class I pathway, the vast majority of antigenic peptides are generated by the proteasome system, a multicatalytic protease complex consisting of constitutive subunits, three of which can be replaced by enzymatically active gamma interferon (IFN-gamma)-inducible subunits, i.e., LMP2, LMP7, and MECL1, to form the so-called immunoproteasomes. Here, we show that steady-state levels of immunoproteasomes are readily formed in response to MCMV infection in the liver. In contrast, the incorporation of immunoproteasome subunits was prevented in MCMV-infected, as well as HCMV-infected, fibroblasts in vitro. Likewise, the expression of the IFN-gamma-inducible proteasome regulator PA28 alpha beta was also impaired in MCMV-infected cells. Both MCMV and HCMV did not alter the constitutive-subunit composition of proteasomes in infected cells. Quantitative assessment of LMP2, MECL1, and LMP7 transcripts revealed that the inhibition of immunoproteasome formation occurred at a pretranscriptional level. Remarkably, a targeted deletion of the MCMV gene M27, encoding an inhibitor of STAT2 that disrupts IFN-gamma receptor signaling, largely restored transcription and protein expression of immunoproteasome subunits in infected cells. While CMV block peptide transport and MHC class I assembly by posttranslational strategies, immunoproteasome assembly, and thus the repertoire of proteasomal peptides, is controlled by pretranscriptional mechanisms. We hypothesize that the blockade of immunoproteasome formation has considerable consequences for shaping the CD8(+)-T cell repertoire during the effector phase of the immune response. PMID- 14747546 TI - Small internal deletions in the human cytomegalovirus IE2 gene result in nonviable recombinant viruses with differential defects in viral gene expression. AB - The human cytomegalovirus (HCMV) IE2 86-kDa protein is a key viral transactivator and an important regulator of HCMV infections. We used the HCMV genome cloned as a bacterial artificial chromosome (BAC) to construct four HCMV mutants with disruptions in regions of IE2 86 that are predicted to be important for its transactivation and autoregulatory functions. Three of these mutants have mutations that remove amino acids 356 to 359, 427 to 435, and 505 to 511, which disrupts a region of IE2 86 implicated in the activation of HCMV early promoters, a predicted zinc finger domain, and a putative helix-loop-helix motif, respectively, while the fourth carries three arginine-to-alanine substitution mutations in the region of amino acids 356 to 359. The resulting recombinant viruses are not viable, and by using quantitative real-time reverse transcription PCR and immunofluorescence we have determined the location of the block in their replicative cycles. The IE2 86 Delta 356-359 mutant is able to support early gene expression, as indicated by the presence of UL112-113 transcripts and UL112-113 and UL44 proteins in cells transfected with the mutant BAC. This mutant does not express late genes and behaves nearly indistinguishably from the IE2 86R356/7/9A substitution mutant. Both exhibit detectable upregulation of major immediate early transcripts at early times. The IE2 86 Delta 427-435 and IE2 86 Delta 505 511 recombinant viruses do not activate the early genes examined and are defective in repression of the major immediate-early promoter. These two mutants also induce the expression of selected delayed early (UL89) and late genes at early times in the infection. We conclude that these three regions of IE2 86 are necessary for productive infections and for differential control of downstream viral gene expression. PMID- 14747548 TI - Chimeric human immunodeficiency virus type 1 (HIV-1) virions containing HIV-2 or simian immunodeficiency virus Nef are resistant to cyclosporine treatment. AB - The viral protein Nef and the cellular factor cyclophilin A are both required for full infectivity of human immunodeficiency virus type 1 (HIV-1) virions. In contrast, HIV-2 and simian immunodeficiency virus (SIV) do not incorporate cyclophilin A into virions or need it for full infectivity. Since Nef and cyclophilin A appear to act in similar ways on postentry events, we determined whether chimeric HIV-1 virions that contained either HIV-2 or SIV Nef would have a direct effect on cyclophilin A dependence. Our results show that chimeric HIV-1 virions containing either HIV-2 or SIV Nef are resistant to treatment by cyclosporine and enhance the infectivity of virions with mutations in the cyclophilin A binding loop of Gag. Amino acids at the C terminus of HIV-2 and SIV are necessary for inducing cyclosporine resistance. However, transferring these amino acids to the C terminus of HIV-1 Nef is insufficient to induce cyclosporine resistance in HIV-1. These results suggest that HIV-2 and SIV Nef are able to compensate for the need for cyclophilin A for full infectivity and that amino acids present at the C termini of these proteins are important for this function. PMID- 14747549 TI - Generation of high-yielding influenza A viruses in African green monkey kidney (Vero) cells by reverse genetics. AB - Influenza A viruses are the cause of annual epidemics of human disease with occasional outbreaks of pandemic proportions. The zoonotic nature of the disease and the vast viral reservoirs in the aquatic birds of the world mean that influenza will not easily be eradicated and that vaccines will continue to be needed. Recent technological advances in reverse genetics methods and limitations of the conventional production of vaccines by using eggs have led to a push to develop cell-based strategies to produce influenza vaccine. Although cell-based systems are being developed, barriers remain that need to be overcome if the potential of these systems is to be fully realized. These barriers include, but are not limited to, potentially poor reproducibility of viral rescue with reverse genetics systems and poor growth kinetics and yields. In this study we present a modified A/Puerto Rico/8/34 (PR8) influenza virus master strain that has improved viral rescue and growth properties in the African green monkey kidney cell line, Vero. The improved properties were mediated by the substitution of the PR8 NS gene for that of a Vero-adapted reassortant virus. The Vero growth kinetics of viruses with H1N1, H3N2, H6N1, and H9N2 hemagglutinin and neuraminidase combinations rescued on the new master strain were significantly enhanced in comparison to those of viruses with the same combinations rescued on the standard PR8 master strain. These improvements pave the way for the reproducible generation of high-yielding human and animal influenza vaccines by reverse genetics methods. Such a means of production has particular relevance to epidemic and pandemic use. PMID- 14747550 TI - African swine fever virus multigene family 360 and 530 genes affect host interferon response. AB - African swine fever virus (ASFV) multigene family 360 and 530 (MGF360/530) genes affect viral growth in macrophage cell cultures and virulence in pigs (L. Zsak, Z. Lu, T. G. Burrage, J. G. Neilan, G. F. Kutish, D. M. Moore, and D. L. Rock, J. Virol. 75:3066-3076, 2001). The mechanism by which these novel genes affect virus host interactions is unknown. To define MGF360/530 gene function, we compared macrophage transcriptional responses following infection with parental ASFV (Pr4) and an MGF360/530 deletion mutant (Pr4 Delta 35). A swine cDNA microarray containing 7,712 macrophage cDNA clones was used to compare the transcriptional profiles of swine macrophages infected with Pr4 and Pr4 Delta 35 at 3 and 6 h postinfection (hpi). While at 3 hpi most (7,564) of the genes had similar expression levels in cells infected with either virus, 38 genes had significantly increased (>2.0-fold, P < 0.05) mRNA levels in Pr4 Delta 35-infected macrophages. Similar up-regulation of these genes was observed at 6 hpi. Viral infection was required for this induced transcriptional response. Most Pr Delta 35 up-regulated genes were part of a type I interferon (IFN) response or were genes that are normally induced by double-stranded RNA and/or viral infection. These included monocyte chemoattractant protein, transmembrane protein 3, tetratricopeptide repeat protein 1, a ubiquitin-like 17-kDa protein, ubiquitin-specific protease ISG43, an RNA helicase DEAD box protein, GTP-binding MX protein, the cytokine IP 10, and the PKR activator PACT. Differential expression of IFN early-response genes in Pr4 Delta 35 relative to Pr4 was confirmed by Northern blot analysis and real-time PCR. Analysis of IFN-alpha mRNA and secreted IFN-alpha levels at 3, 8, and 24 hpi revealed undetectable IFN-alpha in mock- and Pr4-infected macrophages but significant IFN-alpha levels at 24 hpi in Pr4 Delta 35-infected macrophages. The absence of IFN-alpha in Pr4-infected macrophages suggests that MGF360/530 genes either directly or indirectly suppress a type I IFN response. An inability to suppress host type I IFN responses may account for the growth defect of Pr4 Delta 35 in macrophages and its attenuation in swine. PMID- 14747551 TI - The influenza B virus nonstructural NS1 protein is essential for efficient viral growth and antagonizes beta interferon induction. AB - We analyzed the functions of the influenza B virus nonstructural NS1-B protein, both by utilizing a constructed mutant virus (Delta NS1-B) lacking the NS1 gene and by testing the activities of the protein when expressed in cells. The mutant virus replicated to intermediate levels in 6-day-old embryonated chicken eggs that contain an immature interferon (IFN) system, whereas older eggs did not support viral propagation to a significant extent. The Delta NS1-B virus was a substantially stronger inducer of beta IFN (IFN-beta) transcripts in human lung epithelial cells than the wild type, and furthermore, transiently expressed NS1-B protein efficiently inhibited virus-dependent activation of the IFN-beta promoter. Interestingly, replication of the Delta NS1-B knockout virus was attenuated by more than 4 orders of magnitude in tissue culture cells containing or lacking functional IFN-alpha/beta genes. These findings show that the NS1-B protein functions as a viral IFN antagonist and indicate a further requirement of this protein for efficient viral replication that is unrelated to blocking IFN effects. PMID- 14747552 TI - Glycine decarboxylase mediates a postbinding step in duck hepatitis B virus infection. AB - Envelope protein precursors of many viruses are processed by a basic endopeptidase to generate two molecules, one for receptor binding and the other for membrane fusion. Such a cleavage event has not been demonstrated for the hepatitis B virus family. Two binding partners for duck hepatitis B virus (DHBV) pre-S envelope protein have been identified. Duck carboxypeptidase D (DCPD) interacts with the full-length pre-S protein and is the DHBV docking receptor, while duck glycine decarboxylase (DGD) has the potential to bind several deletion constructs of the pre-S protein in vitro. Interestingly, DGD but not DCPD expression was diminished following prolonged culture of primary duck hepatocytes (PDH), which impaired productive DHBV infection. Introduction of exogenous DGD promoted formation of protein-free viral genome, suggesting restoration of several early events in viral life cycle. Conversely, blocking DGD expression in fresh PDH by antisense RNA abolished DHBV infection. Moreover, addition of DGD antibodies soon after virus binding reduced endogenous DGD protein levels and impaired production of covalently closed circular DNA, the template for DHBV gene expression and genome replication. Our findings implicate this second pre-S binding protein as a critical cellular factor for productive DHBV infection. We hypothesize that DCPD, a molecule cycling between the cell surface and the trans Golgi network, targets DHBV particles to the secretary pathway for proteolytic cleavage of viral envelope protein. DGD represents the functional equivalent of other virus receptors in its interaction with processed viral particles. PMID- 14747553 TI - Reovirus nonstructural protein mu NS recruits viral core surface proteins and entering core particles to factory-like inclusions. AB - Mammalian reoviruses are thought to assemble and replicate within cytoplasmic, nonmembranous structures called viral factories. The viral nonstructural protein mu NS forms factory-like globular inclusions when expressed in the absence of other viral proteins and binds to the surfaces of the viral core particles in vitro. Given these previous observations, we hypothesized that one or more of the core surface proteins may be recruited to viral factories through specific associations with mu NS. We found that all three of these proteins--lambda 1, lambda 2, and sigma 2--localized to factories in infected cells but were diffusely distributed through the cytoplasm and nucleus when each was separately expressed in the absence of other viral proteins. When separately coexpressed with mu NS, on the other hand, each core surface protein colocalized with mu NS in globular inclusions, supporting the initial hypothesis. We also found that lambda 1, lambda 2, and sigma 2 each localized to filamentous inclusions formed upon the coexpression of mu NS and mu 2, a structurally minor core protein that associates with microtubules. The first 40 residues of mu NS, which are required for association with mu 2 and the RNA-binding nonstructural protein sigma NS, were not required for association with any of the three core surface proteins. When coexpressed with mu 2 in the absence of mu NS, each of the core surface proteins was diffusely distributed and displayed only sporadic, weak associations with mu 2 on filaments. Many of the core particles that entered the cytoplasm of cycloheximide-treated cells following entry and partial uncoating were recruited to inclusions of mu NS that had been preformed in those cells, providing evidence that mu NS can bind to the surfaces of cores in vivo. These findings expand a model for how viral and cellular components are recruited to the viral factories in infected cells and provide further evidence for the central but distinct roles of viral proteins mu NS and mu 2 in this process. PMID- 14747554 TI - Lytic induction therapy for Epstein-Barr virus-positive B-cell lymphomas. AB - A novel therapy for Epstein-Barr virus (EBV)-positive tumors involves the intentional induction of the lytic form of EBV infection combined with ganciclovir (GCV) treatment. Virally encoded kinases (thymidine kinase and BGLF4) which are expressed only during the lytic form of infection convert GCV (a nucleoside analogue) into its active, cytotoxic form. However, tightly latent EBV infection in B cells has made it difficult to identify drugs that can be used clinically to induce lytic viral infection in B-cell lymphomas. Here we demonstrate that gemcitabine and doxorubicin (but not 5-azacytidine, cis platinum, or 5-fluorouracil) induce lytic EBV infection in EBV-transformed B cells in vitro and in vivo. Gemcitabine and doxorubicin both activated transcription from the promoters of the two viral immediate-early genes, BZLF1 and BRLF1, in EBV-negative B cells. This effect required the EGR-1 motif in the BRLF1 promoter and the CRE (ZII) and MEF-2D (ZI) binding sites in the BZLF1 promoter. GCV enhanced cell killing by gemcitabine or doxorubicin in lymphoblastoid cells transformed with wild-type EBV, but not in lymphoblastoid cells transformed by a mutant virus (with a deletion in the BZLF1 immediate-early gene) that is unable to enter the lytic form of infection. Most importantly, the combination of gemcitabine or doxorubicin and GCV was significantly more effective for the inhibition of EBV-driven lymphoproliferative disease in SCID mice than chemotherapy alone. In contrast, the combination of zidovudine and gemcitabine was no more effective than gemcitabine alone. These results suggest that the addition of GCV to either gemcitabine- or doxorubicin-containing chemotherapy regimens may enhance the therapeutic efficacy of these drugs for EBV driven lymphoproliferative disease in patients. PMID- 14747555 TI - Formation of nuclear foci of the herpes simplex virus type 1 regulatory protein ICP4 at early times of infection: localization, dynamics, recruitment of ICP27, and evidence for the de novo induction of ND10-like complexes. AB - Herpes simplex virus type 1 (HSV-1) has an intricate association with cellular nuclear structures known as ND10 or promyelocytic leukemia protein (PML) nuclear bodies. Parental viral genomes initially become juxtaposed to ND10, and then viral replication compartments develop from the ND10-associated genomes. Viral immediate-early (IE) regulatory protein ICP0 colocalizes with ND10 and then induces the degradation of critical ND10 component protein PML and therefore the release and dispersal of other ND10 proteins. The IE transcriptional regulatory protein ICP4 also forms foci at early times of infection, many of which are juxtaposed to ND10 and later develop into replication compartments, indicating that at least some of the initial ICP4 foci contain parental viral genomes. Here we report that the ICP4 foci also contain ICP27 and that their formation occurs extremely rapidly at locations just inside the nuclear envelope. By examining developing plaques or thinly seeded cells infected at high multiplicity, we found evidence to suggest that at least some of the ND10-viral nucleoprotein complex association could be attributed to de novo formation of ND10-like structures in response to incoming viral genomes. The ICP4 complexes associated efficiently with ND10 in cells infected with an ICP0-null mutant virus at high but not at low multiplicity, and the degree of association was reduced by the proteasome inhibitor MG132. Therefore, the interaction between viral nucleoprotein complexes and ND10 is in part due to a dynamic response by the cell. This response is modulated by functional ICP0, and cells that are productively or nonproductively infected in the absence of functional ICP0 can be distinguished by the relative locations of ICP4 foci and ND10 proteins. PMID- 14747556 TI - The K1 protein of Kaposi's sarcoma-associated herpesvirus activates the Akt signaling pathway. AB - Kaposi's sarcoma-associated herpesvirus (KSHV) has been implicated in Kaposi's sarcoma, as well as in primary effusion lymphoma and multicentric Castleman's disease. The K1 protein of KSHV has been shown to induce cellular transformation and focus formation and to deregulate B-lymphocyte signaling pathways by functionally mimicking the activated B-cell receptor complex. Here we show that expression of K1 in B lymphocytes targets the phosphatidylinositol-3 kinase pathway, leading to the activation of the Akt kinase and the inhibition of the phosphatase PTEN. We also demonstrate that activation of Akt by the K1 protein leads to the phosphorylation and inhibition of members of the forkhead (FKHR) transcription factor family, which are key regulators of cell cycle progression and apoptosis. We demonstrate that K1 can inhibit apoptosis induced by the FKHR proteins and by stimulation of the Fas receptor. Our observations suggest that the K1 viral protein promotes cell survival pathways and may contribute to KSHV pathogenesis by preventing virally infected cells from undergoing apoptosis prematurely. PMID- 14747557 TI - Bending of adenovirus origin DNA by nuclear factor I as shown by scanning force microscopy is required for optimal DNA replication. AB - Nuclear factor I (NFI) is a transcription factor that binds to the adenovirus type 5 (Ad5) origin of replication and recruits the adenovirus DNA polymerase, thereby stimulating initiation of DNA replication in vitro. Using scanning force microscopy, we demonstrate that NFI induces a 60 degrees bend upon binding to the origin. The A/T-rich region preceding the core recognition sequence of NFI influences the DNA bend angle, since substitution of A/T base pairs by G/C base pairs severely decreases bending. Mutations in the A/T-rich region do not affect binding of NFI to DNA. However, mutations that reduce the protein-induced bend lead to a loss of NFI-stimulated replication, indicating that DNA bending is functionally important. In contrast, basal initiation or DNA binding of the polymerase is not impaired by these origin mutations. We conclude that binding of NFI to the Ad5 origin causes structural changes in DNA that are essential for the stimulatory function of NFI in replication. We propose that NFI-induced origin bending facilitates the assembly of a functional initiation complex. PMID- 14747558 TI - Reversible inhibition of spreading of in vitro infection and imbalance of viral protein accumulation at low pH in viral hemorrhagic septicemia rhabdovirus, a salmonid rhabdovirus. AB - The inhibition of viral hemorrhagic septicemia rhabdovirus (VHSV) in vitro infection by pHs of <7 (low pH) has been previously reported. Nevertheless, the details of the mechanism underlying this effect remain obscure. We present evidence showing that low-pH inhibition occurs during a viral postadsorption step. Thus, while VHSV bound, replicated within single cells, and presented its G protein on the membranes of infected cells at both low and physiological pHs, both cell-to-cell spreading of infection (as estimated by the appearance of foci of infected cells) and fusion (as estimated by a syncytium assay) were inhibited by this low pH. The decreased VHSV titers and the inhibition of both cell-to-cell spreading of infection and fusion could be reversed by adjusting the pH to 7.5 at any time during infection. This effect should be taken into account to avoid false negatives in the diagnosis of VHSV by cell culture. On the other hand, the cell-to-cell spreading of infection at pH 7.5 could be stopped at any time by reducing the pH to 6.5. Since at low pH there were changes in the protein G conformation and smaller and imbalanced amounts of N with respect to M1, M2, and G viral proteins, alterations of the assembly and/or budding of VHSV are most probably involved in the absence of newly released infective virions. PMID- 14747559 TI - Identification of antigenic regions of duck hepatitis B virus core protein with antibodies elicited by DNA immunization and chronic infection. AB - The induction of humoral response in ducks by DNA-based immunization against duck hepatitis B virus (DHBV) core protein (DHBc) was investigated. In addition, the amino acid specificity of the induced response was compared by using peptide scanning to that elicited either by protein immunization or during chronic DHBV infection. Immunization of ducks with a plasmid expressing DHBc protein led to the induction of a long-lasting antibody response able to specifically recognize viral protein in chronically infected duck livers. Peptide scanning analysis of anti-DHBc response induced during chronic DHBV infection allowed us to identify six major antigenic regions (AR1 to AR6). The reactivity spectrum of duck sera elicited by protein immunization appeared narrower and was restricted to only four of these antigenic regions in spite of higher anti-DHBc antibody titers. Interestingly, anti-DHBc antibodies induced by DNA-based immunization recognized five of six antigenic regions, and the epitope pattern was broader and more closely related to that observed in chronic viral infections. To gain more insight into the location of antigenic regions, we built a three-dimensional (3 D) model of DHBc protein based on human and duck core sequence alignment data and the HBc 3-D crystal structure. The results suggest that two identified antigenic regions (AR2, amino acids [aa] (64)T-P(84), and AR5, aa (183)A-R(210)) are located at positions on the protein surface equivalent to those of the two HBc major epitopes. Moreover, we identified another antigenic region (AR3, aa (99)I I(112)) that was recognized by all sera from chronically infected, DNA- or protein-immunized ducks within the large 45-aa insertion in DHBc protein, suggesting that this region, which lacks HBc, is externally exposed. PMID- 14747560 TI - Comparative genomic analysis of hyperthermophilic archaeal Fuselloviridae viruses. AB - The complete genome sequences of two Sulfolobus spindle-shaped viruses (SSVs) from acidic hot springs in Kamchatka (Russia) and Yellowstone National Park (United States) have been determined. These nonlytic temperate viruses were isolated from hyperthermophilic Sulfolobus hosts, and both viruses share the spindle-shaped morphology characteristic of the Fuselloviridae family. These two genomes, in combination with the previously determined SSV1 genome from Japan and the SSV2 genome from Iceland, have allowed us to carry out a phylogenetic comparison of these geographically distributed hyperthermal viruses. Each virus contains a circular double-stranded DNA genome of approximately 15 kbp with approximately 34 open reading frames (ORFs). These Fusellovirus ORFs show little or no similarity to genes in the public databases. In contrast, 18 ORFs are common to all four isolates and may represent the minimal gene set defining this viral group. In general, ORFs on one half of the genome are colinear and highly conserved, while ORFs on the other half are not. One shared ORF among all four genomes is an integrase of the tyrosine recombinase family. All four viral genomes integrate into their host tRNA genes. The specific tRNA gene used for integration varies, and one genome integrates into multiple loci. Several unique ORFs are found in the genome of each isolate. PMID- 14747561 TI - Comparative study of adaptive molecular evolution in different human immunodeficiency virus groups and subtypes. AB - Molecular adaptation, as characterized by the detection of positive selection, was quantified in a number of genes from different human immunodeficiency virus type 1 (HIV-1) group M subtypes, group O, and an HIV-2 subtype using the codon based maximum-likelihood method of Yang and coworkers (Z. H. Yang, R. Nielsen, N. Goldman, and A. M. K. Pedersen, Genetics 155:431-449, 2000). The env gene was investigated further since it exhibited the strongest signal for positive selection compared to those of the other two major HIV genes (gag and pol). In order to investigate the pattern of adaptive evolution across env, the location and strength of positive selection in different HIV-1 sequence alignments was compared. The number of sites having a significant probability of being positively selected varied among these different alignment data sets, ranging from 25 in HIV-1 group M subtype A to 40 in HIV-1 group O. Strikingly, there was a significant tendency for positively selected sites to be located at the same position in different HIV-1 alignments, ranging from 10 to 16 shared sites for the group M intersubtype comparisons and from 6 to 8 for the group O to M comparisons, suggesting that all HIV-1 variants are subject to similar selective forces. As the host immune response is believed to be the dominant driving force of adaptive evolution in HIV, this result would suggest that the same sites are contributing to viral persistence in diverse HIV infections. Thus, the positions of the positively selected sites were investigated in reference to the inferred locations of different epitope types (antibody, T helper, and cytotoxic T lymphocytes) and the positions of N and O glycosylation sites. We found a significant tendency for positively selected sites to fall outside T-helper epitopes and for positively selected sites to be strongly associated with N glycosylation sites. PMID- 14747562 TI - Large-scale identification of disease genes involved in acute myeloid leukemia. AB - Acute myeloid leukemia (AML) is a heterogeneous group of diseases in which chromosomal aberrations, small insertions or deletions, or point mutations in certain genes have profound consequences for prognosis. However, the majority of AML patients present without currently known genetic defects. Retroviral insertion mutagenesis in mice has become a powerful tool for identifying new disease genes involved in the pathogenesis of leukemia and lymphoma. Here we have used the Graffi-1.4 strain of murine leukemia virus, which causes predominantly AML, in a screen to identify novel genes involved in the pathogenesis of this disease. We report 79 candidate disease genes in common integration sites (CISs) and 15 genes whose family members previously were found to be affected in other studies. The majority of the identified sequences (60%) were not found in lymphomas and monocytic leukemias in previous screens, suggesting a specific involvement in AML. Although most of the virus integrations occurred in or near the 5' or 3' ends of the genes, suggesting deregulation of gene expression as a consequence of virus integration, 18 CISs were located exclusively within the genes, conceivably causing gene disruption. PMID- 14747564 TI - Members of a novel family of mammalian protein kinases complement the DNA negative phenotype of a vaccinia virus ts mutant defective in the B1 kinase. AB - Temperature-sensitive (ts) mutants of vaccinia virus defective in the B1 kinase demonstrate a conditionally lethal defect in DNA synthesis. B1 is the prototypic member of a new family of protein kinases (vaccinia virus-related kinases, or VRK) that possess distinctive B1-like sequence features within their catalytic motifs (R. J. Nichols and P. Traktman, J. Biol. Chem., in press). Given the striking sequence similarity between B1 and the VRK enzymes, we proposed that they might share overlapping substrate specificity. We therefore sought to determine whether the human and mouse VRK1 enzymes (hVRK1 and mVRK1, respectively) could complement a B1 deficiency in vivo. Recombinant ts2 viruses expressing hVRK1, mVRK1, or wild-type B1 were able to synthesize viral DNA at high temperature, but those expressing the more distantly related human casein kinase 1 alpha 2 could not. Complementation required the enzymatic activity of hVRK1, since a catalytically inactive allele of hVRK1 was unable to confer a temperature-insensitive phenotype. Interestingly, rescue of viral DNA synthesis was not coupled to the ability to phosphorylate H5, the only virus-encoded protein shown to be a B1 substrate in vivo. Expression of hVRK1 during nonpermissive ts2 infections restored virus production and plaque formation, whereas expression of mVRK1 resulted in an intermediate level of rescue. Taken together, these observations indicate that enzymatically active cellular VRK1 kinases can perform the function(s) of B1 required for genome replication, most likely due to overlapping specificity for cellular and/or viral substrates. PMID- 14747563 TI - Epstein-Barr virus nuclear antigen 3C regulates cyclin A/p27 complexes and enhances cyclin A-dependent kinase activity. AB - Epstein-Barr virus (EBV) nuclear antigen 3C (EBNA3C) is essential for primary B cell transformation. In this report we show that cyclin A, an activator of S phase progression, bound tightly to EBNA3C. EBNA3C interacted with cyclin A in vitro and associated with cyclin A complexes in EBV-transformed lymphoblastoid cell lines. Importantly, EBNA3C stimulated cyclin A-dependent kinase activity and rescued p27-mediated inhibition of cyclin A/Cdk2 kinase activity by decreasing the molecular association between cyclin A and p27 in cells. Additionally, phosphorylation of the retinoblastoma protein, a major regulator of cell cycle progression, was enhanced both in vitro and in vivo in the presence of EBNA3C. Cyclin A interacted with a region of the carboxy terminus of EBNA3C, shown to be important both for stimulation of cyclin A-dependent kinase activity and for cell cycle progression. This provides the first evidence of an essential EBV latent antigen's directly targeting a cell cycle regulatory protein and suggests a novel mechanism by which EBV deregulates the mammalian cell cycle, which is of critical importance in B-cell transformation. PMID- 14747565 TI - Lv2, a novel postentry restriction, is mediated by both capsid and envelope. AB - The characterization of restrictions to lentivirus replication in cells identifies critical steps in the viral life cycle and potential therapeutic targets. We previously reported that a human immunodeficiency virus type 2 (HIV 2) isolate was restricted to infection in some human cells, which led us to identify a step in the life cycle of HIV-2 detected after reverse transcription but prior to nuclear entry. The block is bypassed with a vesicular stomatitis virus glycoprotein G (VSV-G) envelope (A. McKnight et al., J. Virol. 75:6914 6922, 2001). We hypothesized that, although the restriction is apparent at a post reverse transcription step, the lack of progress results from a failure of the virus to reach a cellular compartment with access to the nucleus. Here we analyzed molecular clones of the restricted virus, MCR, and an unrestricted virus, MCN. Using sequence analysis and gene swapping, we mapped the viral determinants to gag and env. Site-directed mutagenesis identified a single amino acid at position 207 in CA to be responsible for the gag restriction. Pseudotype experiments indicate that this step is also important for the infection of cells by HIV-1. The HIV-1 NL4.3 core is restricted if supplied with a restricted MCR envelope but not with VSV-G. Also the NL4.3 envelope rescues the restricted core of HIV-2 MCR. Abrogation experiments with MLV demonstrate that the restriction is distinct from Fv1/Ref1/Lv1. We propose that this represents a new lentiviral restriction, Lv2. Thus, the envelope and capsid of HIV act to ensure that the virus is delivered into an appropriate cellular compartment that allows postentry events in viral replication to proceed efficiently. PMID- 14747566 TI - Generation of recombinant human parainfluenza virus type 1 vaccine candidates by importation of temperature-sensitive and attenuating mutations from heterologous paramyxoviruses. AB - Human parainfluenza virus type 1 (HPIV1) is a significant cause of respiratory tract disease in infants and young children for which a vaccine is needed. In the present study, we sought to attenuate HPIV1 by the importation of one or more known attenuating point mutations from heterologous paramyxoviruses into homologous sites in HPIV1. The introduced mutations were derived from three attenuated paramyxoviruses: (i) HPIV3cp45, a live-attenuated HPIV3 vaccine candidate containing multiple attenuating mutations; (ii) the respiratory syncytial virus cpts530 with an attenuating mutation in the L polymerase protein; and (iii) a murine PIV1 (MPIV1) attenuated by a mutation in the accessory C protein. Recombinant HPIV1 (rHPIV1) mutants bearing a single imported mutation in C, any of three different mutations in L, or a pair of mutations in F exhibited a 100-fold or greater reduction in replication in the upper or lower respiratory tract of hamsters. Both temperature-sensitive (ts) (mutations in the L and F proteins) and non-ts (the mutation in the C protein) attenuating mutations were identified. rHPIV1 mutants containing a combination of mutations in L were generated that were more attenuated than viruses bearing the individual mutations, showing that the systematic accretion of mutations can yield progressive increases in attenuation. Hamsters immunized with rHPIV1 mutants bearing one or two mutations developed neutralizing antibodies and were resistant to challenge with wild-type HPIV1. Thus, importation of attenuating mutations from heterologous viruses is an effective means for rapidly identifying mutations that attenuate HPIV1 and for generating live-attenuated HPIV1 vaccine candidates. PMID- 14747567 TI - Codon substitution mutations at two positions in the L polymerase protein of human parainfluenza virus type 1 yield viruses with a spectrum of attenuation in vivo and increased phenotypic stability in vitro. AB - The Y942H and L992F temperature-sensitive (ts) and attenuating amino acid substitution mutations, previously identified in the L polymerase of the HPIV3cp45 vaccine candidate, were introduced into homologous positions of the L polymerase of recombinant human parainfluenza virus type 1 (rHPIV1). In rHPIV1, the Y942H mutation specified the ts phenotype in vitro and the attenuation (att) phenotype in hamsters, whereas the L992F mutation specified neither phenotype. Each of these codon mutations was generated by a single nucleotide substitution and therefore had the potential to readily revert to a codon specifying the wild type amino acid residue. We introduced alternative amino acid assignments at codon 942 or 992 as a strategy to increase genetic stability and to generate mutants that exhibit a range of attenuation. Twenty-three recombinants with codon substitutions at position 942 or 992 of the L protein were viable. One highly ts and att mutant, the Y942A virus, which had a difference of three nucleotides from the codon encoding a wild-type tyrosine, also possessed a high level of genetic and phenotypic stability upon serial passage in vitro at restrictive temperatures compared to that of the parent Y942H virus, which possessed a single nucleotide substitution. We obtained mutants with substitutions at position 992 that, in contrast to the L992F virus, possessed the ts and att phenotypes. These findings identify the use of alternative codon substitution mutations as a method that can be used to generate candidate vaccine viruses with increased genetic stability and/or a modified level of attenuation. PMID- 14747568 TI - Discrete domains within the rotavirus VP5* direct peripheral membrane association and membrane permeability. AB - Cleavage of the rotavirus spike protein, VP4, is required for rotavirus-induced membrane permeability and viral entry into cells. The VP5* cleavage product selectively permeabilizes membranes and liposomes and contains an internal hydrophobic domain that is required for membrane permeability. Here we investigate VP5* domains (residues 248 to 474) that direct membrane binding. We determined that expressed VP5 fragments containing residues 248 to 474 or 265 to 474, including the internal hydrophobic domain, bind to cellular membranes but are not present in Triton X-100-resistant membrane rafts. Expressed VP5 partitions into aqueous but not detergent phases of Triton X-114, suggesting that VP5 is not integrally inserted into membranes. Since high-salt or alkaline conditions eluted VP5 from membranes, our findings demonstrate that VP5 is peripherally associated with membranes. Interestingly, mutagenesis of residue 394 (W-->R) within the VP5 hydrophobic domain, which abolishes VP5-directed permeability, had no effect on VP5's peripheral membrane association. In contrast, deletion of N-terminal VP5 residues (residues 265 to 279) abolished VP5 binding to membranes. Alanine mutagenesis of two positively charged residues within this domain (residues 274R and 276K) dramatically reduced (>95%) binding of VP5 to membranes and suggested their potential interaction with polar head groups of the lipid bilayer. Mutations in either the VP5 hydrophobic or basic domain blocked VP5-directed permeability of cells. These findings indicate that there are at least two discrete domains within VP5* required for pore formation: an N-terminal basic domain that permits VP5* to peripherally associate with membranes and an internal hydrophobic domain that is essential for altering membrane permeability. These results provide a fundamental understanding of interactions between VP5* and the membrane, which are required for rotavirus entry. PMID- 14747570 TI - Protection of Penaeus monodon against white spot syndrome virus by oral vaccination. AB - White spot syndrome virus (WSSV) occurs worldwide and causes high mortality and considerable economic damage to the shrimp farming industry. No adequate treatments against this virus are available. It is generally accepted that invertebrates such as shrimp do not have an adaptive immune response system such as that present in vertebrates. As it has been demonstrated that shrimp surviving a WSSV infection have higher survival rates upon subsequent rechallenge, we investigated the potential of oral vaccination of shrimp with subunit vaccines consisting of WSSV virion envelope proteins. Penaeus monodon shrimp were fed food pellets coated with inactivated bacteria overexpressing two WSSV envelope proteins, VP19 and VP28. Vaccination with VP28 showed a significant lower cumulative mortality compared to vaccination with bacteria expressing the empty vectors after challenge via immersion (relative survival, 61%), while vaccination with VP19 provided no protection. To determine the onset and duration of protection, challenges were subsequently performed 3, 7, and 21 days after vaccination. A significantly higher survival was observed both 3 and 7 days postvaccination (relative survival, 64% and 77%, respectively), but the protection was reduced 21 days after the vaccination (relative survival, 29%). This suggests that contrary to current assumptions that invertebrates do not have a true adaptive immune system, a specific immune response and protection can be induced in P. monodon. These experiments open up new ways to benefit the WSSV hampered shrimp farming industry. PMID- 14747569 TI - Complete genome sequence of Fer-de-Lance virus reveals a novel gene in reptilian paramyxoviruses. AB - The complete RNA genome sequence of the archetype reptilian paramyxovirus, Fer-de Lance virus (FDLV), has been determined. The genome is 15,378 nucleotides in length and consists of seven nonoverlapping genes in the order 3' N-U-P-M-F-HN-L 5', coding for the nucleocapsid, unknown, phospho-, matrix, fusion, hemagglutinin neuraminidase, and large polymerase proteins, respectively. The gene junctions contain highly conserved transcription start and stop signal sequences and tri nucleotide intergenic regions similar to those of other Paramyxoviridae. The FDLV P gene expression strategy is like that of rubulaviruses, which express the accessory V protein from the primary transcript and edit a portion of the mRNA to encode P and I proteins. There is also an overlapping open reading frame potentially encoding a small basic protein in the P gene. The gene designated U (unknown), encodes a deduced protein of 19.4 kDa that has no counterpart in other paramyxoviruses and has no similarity with sequences in the National Center for Biotechnology Information database. Active transcription of the U gene in infected cells was demonstrated by Northern blot analysis, and bicistronic N-U mRNA was also evident. The genomes of two other snake paramyxovirus genotypes were also found to have U genes, with 11 to 16% nucleotide divergence from the FDLV U gene. Pairwise comparisons of amino acid identities and phylogenetic analyses of all deduced FDLV protein sequences with homologous sequences from other Paramyxoviridae indicate that FDLV represents a new genus within the subfamily Paramyxoviridae. We suggest the name Ferlavirus for the new genus, with FDLV as the type species. PMID- 14747571 TI - Development of a GB virus B marmoset model and its validation with a novel series of hepatitis C virus NS3 protease inhibitors. AB - GB virus B (GBV-B), a flavivirus closely related to HCV, has previously been shown to infect and replicate to high titers in tamarins (Saguinus sp.). This study describes the use of GBV-B infection and replication in the common marmoset (Callithrix jacchus) for the successful development and validation of a surrogate animal model for hepatitis C virus (HCV). Infection of marmosets with GBV-B produced a viremia that peaked at 10(8) to 10(9) genome copies/ml for a period of 40 to 60 days followed by viral clearance at 60 to 80 days postinfection. Passage of the initial tamarin-derived GBV-B in marmosets produced an infectious stock that gave a more reproducible and consistent infection in the marmoset. Titration of the virus stocks in vivo indicated that they contained 1 infectious unit for every 1,000 genome copies. Cultures of primary marmoset hepatocytes were also successfully infected with GBV-B, with high levels of virus detected in supernatants and cells for up to 14 days postinfection. Treatment of GBV-B infected hepatocyte cultures with a novel class of HCV protease inhibitor (pyrrolidine 5,5 trans-lactams) reduced viral levels by more than 2 logs. Treatment of GBV-B-infected marmosets with one such inhibitor resulted in a 3-log drop in serum viral titer over 4 days of therapy. These studies provide the first demonstration of the in vivo efficacy of a small-molecule inhibitor for HCV in an animal model and illustrate the utility of GBV-B as a surrogate animal model system for HCV. PMID- 14747572 TI - Influence of primate lentiviral Vif and proteasome inhibitors on human immunodeficiency virus type 1 virion packaging of APOBEC3G. AB - The Vif protein of human immunodeficiency virus type 1 (HIV-1) is essential for viral evasion of the host antiviral protein APOBEC3G, also known as CEM15. Vif mutant but not wild-type HIV-1 viruses produced in the presence of APOBEC3G have been shown to undergo hypermutations in newly synthesized viral DNA upon infection of target cells, presumably resulting from C-to-U modification during minus-strand viral DNA synthesis. We now report that HIV-1 Vif could induce rapid degradation of human APOBEC3G that was blocked by the proteasome inhibitor MG132. The efficiency of Vif-induced downregulation of APOBEC3G expression depended on the level of Vif expression. A single amino acid substitution in the conserved SLQXLA motif reduced Vif function. Vif proteins from distantly related primate lentiviruses such as SIVagm were unable to suppress the antiviral activity of human APOBEC3G or the packaging of APOBEC3G into HIV-1 Vif mutant virions, due to a lack of interaction with human APOBEC3G. In the presence of the proteasome inhibitor MG132, virion-associated Vif increased dramatically. However, increased virion packaging of Vif did not prevent virion packaging of APOBEC3G when proteasome function was impaired, and the infectivity of these virions was significantly reduced. These results suggest that Vif function is required during virus assembly to remove APOBEC3G from packaging into released virions. Once packaged, virion-associated Vif could not efficiently block the antiviral activity of APOBEC3G. PMID- 14747573 TI - The human immunodeficiency virus type 1 ribosomal frameshifting site is an invariant sequence determinant and an important target for antiviral therapy. AB - Human immunodeficiency virus type 1 (HIV-1) utilizes a distinctive form of gene regulation as part of its life cycle, termed programmed -1 ribosomal frameshifting, to produce the required ratio of the Gag and Gag-Pol polyproteins. We carried out a sequence comparison of 1,000 HIV-1 sequences at the slippery site (UUUUUUA) and found that the site is invariant, which is somewhat surprising for a virus known for its variability. This prompted us to prepare a series of mutations to examine their effect upon frameshifting and viral infectivity. Among the series of mutations were changes of the HIV-1 slippery site to those effectively utilized by other viruses, because such mutations would be anticipated to have a relatively mild effect upon frameshifting. The results demonstrate that any change to the slippery site reduced frameshifting levels and also dramatically inhibited infectivity. Because ribosomal frameshifting is essential for HIV-1 replication and it is surprisingly resistant to mutation, modulation of HIV-1 frameshifting efficiency potentially represents an important target for the development of novel antiviral therapeutics. PMID- 14747574 TI - Mutant PrPSc conformers induced by a synthetic peptide and several prion strains. AB - Gerstmann-Straussler-Scheinker (GSS) disease is a dominantly inherited, human prion disease caused by a mutation in the prion protein (PrP) gene. One mutation causing GSS is P102L, denoted P101L in mouse PrP (MoPrP). In a line of transgenic mice denoted Tg2866, the P101L mutation in MoPrP produced neurodegeneration when expressed at high levels. MoPrP(Sc)(P101L) was detected both by the conformation dependent immunoassay and after protease digestion at 4 degrees C. Transmission of prions from the brains of Tg2866 mice to those of Tg196 mice expressing low levels of MoPrP(P101L) was accompanied by accumulation of protease-resistant MoPrP(Sc)(P101L) that had previously escaped detection due to its low concentration. This conformer exhibited characteristics similar to those found in brain tissue from GSS patients. Earlier, we demonstrated that a synthetic peptide harboring the P101L mutation and folded into a beta-rich conformation initiates GSS in Tg196 mice (29). Here we report that this peptide-induced disease can be serially passaged in Tg196 mice and that the PrP conformers accompanying disease progression are conformationally indistinguishable from MoPrP(Sc)(P101L) found in Tg2866 mice developing spontaneous prion disease. In contrast to GSS prions, the 301V, RML, and 139A prion strains produced large amounts of protease-resistant PrP(Sc) in the brains of Tg196 mice. Our results argue that MoPrP(Sc)(P101L) may exist in at least several different conformations, each of which is biologically active. Such conformations occurred spontaneously in Tg2866 mice expressing high levels of MoPrP(C)(P101L) as well as in Tg196 mice expressing low levels of MoPrP(C)(P101L) that were inoculated with brain extracts from ill Tg2866 mice, with a synthetic peptide with the P101L mutation and folded into a beta-rich structure, or with prions recovered from sheep with scrapie or cattle with bovine spongiform encephalopathy. PMID- 14747576 TI - Evolutionary potential of an RNA virus. AB - RNA viruses are remarkably adaptable to changing environments. This is medically important because it enables pathogenic viruses to escape the immune response and chemotherapy and is of considerable theoretical interest since it allows the investigation of evolutionary processes within convenient time scales. A number of earlier studies have addressed the dynamics of adapting RNA virus populations. However, it has been difficult to monitor the trajectory of molecular changes in RNA genomes in response to selective pressures. To address the problem, we developed a novel in vitro evolution system based on a recombinant double stranded RNA bacteriophage, phi 6, containing a beta-lactamase (bla) gene marker. Carrier-state bacterial cells are resistant to ampicillin, and after several passages, they become resistant to high concentrations of another beta-lactam antibiotic, cefotaxime, due to mutations in the virus-borne bla gene. We monitored the changes in bla cDNAs induced by cefotaxime selection and observed an initial explosion in sequence variants with multiple mutations throughout the gene. After four passages, a stable, homogeneous population of bla sequences containing three specific nonsynonymous mutations was established. Of these, two mutations (E104K and G238S) have been previously reported for beta-lactamases from cefotaxime-resistant bacterial isolates. These results extend our understanding of the molecular mechanisms of viral adaptation and also demonstrate the possibility of using an RNA virus as a vehicle for directed evolution of heterologous proteins. PMID- 14747578 TI - Distribution of hydrophobic residues is crucial for the fusogenic properties of the Ebola virus GP2 fusion peptide. AB - The lipid-destabilizing properties of the N-terminal domain of the GP2 of Ebola virus were investigated. Our results suggest that the domain of Ebola virus needed for fusion is shorter than that previously reported. The fusogenic properties of this domain are related to its oblique orientation at the lipid/water interface owing to an asymmetric distribution of the hydrophobic residues when helical. PMID- 14747575 TI - Analysis of chromatin attachment and partitioning functions of bovine papillomavirus type 1 E2 protein. AB - Recent studies have suggested that the tethering of viral genomes to host cell chromosomes could provide one of the ways to achieve their nuclear retention and partitioning during extrachromosomal maintenance in dividing cells. The data we present here provide firm evidence that the partitioning of the bovine papillomavirus type 1 (BPV1) genome is dependent on the chromatin attachment process mediated by viral E2 protein and its multiple binding sites. On the other hand, the attachment of E2 and the E2-mediated tethering of reporter plasmids to host chromosomes are not necessarily sufficient for efficient partitioning, suggesting that additional E2-dependent activities might be involved in the latter process. The activity of E2 protein in chromatin attachment and partitioning is more sensitive to the point mutations in the N-terminal domain than its transactivation and replication initiation functions. Therefore, at least part of the interactions of the E2 N-terminal domain with its targets during the chromatin attachment and partitioning processes are likely to involve specific receptors not involved in transactivation and replication activities of the protein. The mutational analysis also indicates that the binding of E2 to chromatin is not achieved through interaction of linear N-terminal subsequences of the E2 protein with putative receptors. Instead, the composite surface elements of the N-terminal domain build up the receptor-binding surface of E2. In this regard, the interaction of BPV1 E2 with its chromosomal targets clearly differs from the interactions of LANA1 protein from Kaposi's sarcoma-associated human herpesvirus and EBNA1 from Epstein-Barr virus with their specific receptors. PMID- 14747577 TI - Induction of simian AIDS in infant rhesus macaques infected with CCR5- or CXCR4 utilizing simian-human immunodeficiency viruses is associated with distinct lesions of the thymus. AB - Newborn rhesus macaques were infected with two chimeric simian-human immunodeficiency virus (SHIV) strains which contain unique human immunodeficiency virus type 1 (HIV-1) env genes and exhibit distinct phenotypes. Infection with either the CCR5-specific SHIV(SF162P3) or the CXCR4-utilizing SHIV(SF33A) resulted in clinical manifestations consistent with simian AIDS. Most prominent in this study was the detection of severe thymic involution in all SHIV(SF33A) infected infants, which is very similar to HIV-1-induced thymic dysfunction in children who exhibit a rapid pattern of disease progression. In contrast, SHIV(SF162P3) induced only a minor disruption in thymic morphology. Consistent with the distribution of the coreceptors CXCR4 and CCR5 within the thymus, the expression of SHIV(SF162P3) was restricted to the thymic medulla, whereas SHIV(SF33A) was preferentially detected in the cortex. This dichotomy of tissue tropism is similar to the differential tropism of HIV-1 isolates observed in the reconstituted human thymus in SCID-hu mice. Accordingly, our results show that the SHIV-monkey model can be used for the molecular dissection of cell and tissue tropisms controlled by the HIV-1 env gene and for the analysis of mechanisms of viral immunopathogenesis in AIDS. Furthermore, these findings could help explain the rapid progression of disease observed in some HIV-1-infected children. PMID- 14747579 TI - Antiviral activity of distamycin A against vaccinia virus is the result of inhibition of postreplicative mRNA synthesis. AB - Distamycin A has been described as an inhibitor of the cellular pathogenesis of vaccinia virus in culture. Distamycin is an antibiotic that specifically targets the minor groove of DNA. We show here that distamycin is a potent inhibitor of vaccinia virus replication. Pulse-labeling experiments showed that most major late proteins failed to accumulate in the presence of the antibiotic. We characterized the effect of distamycin on vaccinia virus nucleic acid biosynthesis with the goal of determining the inhibitor's target. Early gene transcription was unaffected. DNA synthesis proceeded at normal rates, but DNA accumulated in large masses in the cytoplasm with no evidence of virion assembly. Transcription from the intermediate class promoter for the I1L gene was partially reduced by distamycin; however, transcription from the intermediate promoters for the three late transcription factor genes was severely inhibited. The accumulation of the late transcripts for the viral F17R and A10L genes also was severely impaired and was shown to be a direct inhibition of late promoter activity. These results indicate that inhibition of postreplicative intermediate and late transcription is the basis for inhibition of vaccinia virus by distamycin and indicate that DNA minor-groove ligands hold promise for effective anti-poxvirus drugs. PMID- 14747581 TI - Cytotoxic T lymphocytes derived from patients with chronic hepatitis C virus infection kill bystander cells via Fas-FasL interaction. AB - The role of Fas-mediated lysis of hepatocytes in hepatitis C virus (HCV)-induced injury is frequently discussed. We therefore analyzed the effect of the number of HCV antigen-expressing cells, the mode of antigen presentation, and the number of cytotoxic T lymphocytes in a coculture system mimicking cellular components of the liver. Here, we show that endogenously processed HCV proteins are capable of inducing bystander killing. We further demonstrate that 0.8 to 1.5% of cells presenting HCV antigens suffice to induce lysis of 10 to 29% of bystander cells, suggesting that the mechanism may be operative at low fractions of infected versus uninfected hepatocytes in vivo. Our data underscore the role of the Fas pathway in HCV-related liver injury and support the exploration of Fas-based treatment strategies for patients with chronic hepatitis C virus infection. PMID- 14747580 TI - The viral E4 protein is required for the completion of the cottontail rabbit papillomavirus productive cycle in vivo. AB - Expression of the papillomavirus E4 protein correlates with the onset of viral DNA amplification. Using a mutant cottontail rabbit papillomavirus (CRPV) genome incapable of expressing the viral E4 protein, we have shown that E4 is required for the productive stage of the CRPV life cycle in New Zealand White and cottontail rabbits. In these lesions, E4 was not required for papilloma development, but the onset of viral DNA amplification and L1 expression were abolished. Viral genome amplification was partially restored when mutant genomes able to express longer forms of E4 were used. These findings suggest that efficient amplification of the CRPV genome is dependent on the expression of a full-length CRPV E4 protein. PMID- 14747583 TI - Inactivation of transmissible spongiform encephalopathy (prion) agents by environ LpH. AB - Agents causing transmissible spongiform encephalopathy (TSE) diseases are resistant to inactivation by several conventional decontamination methods. Using an animal bioassay, we compared the TSE agent disinfectant efficacy of a commercially available product referred to alternatively as LpH-SE, LpH-AG, or LpH-st to that of a similarly named but differently formulated product, Environ LpH, which was found to be an effective TSE agent disinfectant in a previous study. Here, we found LpH-SE to be at least 10(4)-fold to 10(5)-fold less effective than Environ LpH. PMID- 14747582 TI - Phylogenetic analyses of Texas isolates indicate an evolving subtype of the clade B feline immunodeficiency viruses. AB - Rigorous phylogenetic analyses were used to compare the nucleotide sequences of feline immunodeficiency virus strains isolated from Texas and throughout the world. The envelope V3-V4 sequences and capsid gene of the Texas isolates formed a cluster between subtypes B and E. Statistical comparisons with other published sequences confirmed that the Texas group is a unique cluster, possibly a new subtype, arising from subtype B. PMID- 14747585 TI - Regulating health in Europe. PMID- 14747586 TI - Addressing health inequalities in the United Kingdom: a case study. AB - Health inequalities research has a long history in the United Kingdom, and the development of government policies that are intended to explicitly address the existing health inequalities has been gathering pace since the Labour Party returned to power in 1997. In this paper, using the influential Acheson Report as a reference point, one of us (D.N.) describes how health inequalities policies have been developed, and the other (A.O.) assesses how, ideally, such policies ought to be developed. Although progress in the development of health inequalities policies has been made, the policies, and the evidence that has informed them, have been less than ideal. PMID- 14747587 TI - Socio-economic position and health: what you observe depends on how you measure it. AB - BACKGROUND: A number of different socio-economic classifications have been used in relation to health in the United Kingdom. The aim of this study was to compare the predictive power of different socio-economic classifications in relation to a range of health measures. METHODS: A postal questionnaire was sent to a random sample of adults in the West of Scotland (sampling from 1997 electoral roll, response rate 50 percent achieved sample 2,867). RESULTS: Associations between social position and health vary by socio-economic classification, health measure and gender. Limiting long-standing illness is more socially patterned than recent illness; income, Registrar General Social Class, housing tenure and car access are more predictive of health than the new National Statistics Socio Economic Classification; and men show steeper socio-economic gradients than women. CONCLUSION: Although there is a consistent picture of poorer health among more disadvantaged groups, however measured, in seeking to explain and reduce social inequalities in health we need to take a more differentiated approach that does not assume equivalence among social classifications and health measures. PMID- 14747588 TI - Urban-rural differences in self-reported limiting long-term illness in Scotland. AB - BACKGROUND: Previous research suggests that there are significant differences in health between urban and rural areas. The aim of this study is to describe the pattern and magnitude of urban-rural variation in health in Scotland and to examine the factors associated with health inequalities in urban and rural areas. METHODS: The data used in this study were limiting long-term illness (LLTI) and socio-economic data collected by the 1991 Census. A rurality indicator was created using Scottish Household Survey rurality classifications. Multilevel Poisson regression modelling was carried out with LLTI as a health indicator for each type of rurality within Scotland. A variety of socio-economic factors were investigated for each rurality. RESULTS: Areas with the highest Standardized Illness Ratios (SIRs) (>125) are predominantly urban whereas the lowest SIRs (<75) are found in both urban and rural areas. Rural communities are more heterogeneous than urban areas in terms of their social make-up with relation to health; however, when these areas are split according to minor road length and different socio-economic factors are added, the model fit for each new model is improved and the reduction in total variation is comparable with that of the urban models. CONCLUSION: These findings suggest that rural areas should not be treated as a homogeneous group but should be subdivided into rural types. PMID- 14747589 TI - Inhalation sedation with nitrous oxide as an alternative to dental general anaesthesia for children. AB - This review paper examines (using systematic methodology) the evidence for the use of inhalation sedation (IHS) instead of dental general anaesthesia (DGA) for dental treatment. It finds that this is an area of healthcare lacking high quality clinical evidence (i.e. derived from randomized controlled trials). However, evidence from seven case series studies (level of evidence 3) of variable quality and design is examined. Those studies suggest that IHS is effective for a large proportion (83-97 percent) of selected subgroups of children who would have otherwise required DGA. This may represent 45-64 percent of all children who are referred for DGA. There is a remarkable degree of consistency between all studies in the reported treatment effectiveness of IHS, despite differences in design and populations treated. IHS is particularly suitable for orthodontic treatment, for older children, and for children requiring no more than four extractions. Morbidity associated with IHS is minor and infrequent, and user satisfaction is high, or higher compared with DGA. Comparing with DGA, IHS requires significantly longer time per episode and more treatment sessions per patient. In teaching dental hospitals, staffing costs for IHS are estimated to be cheaper by about a third compared with outpatient DGA. Indications for further areas of research are made. PMID- 14747590 TI - Cancer patients' awareness about their diagnosis: a population-based study. AB - BACKGROUND: The aim of the study was to evaluate Norwegian cancer patients' awareness of their prior cancer diagnosis in a general population-based study. METHODS: A cross-sectional population-based study of cancer patients' responses to the index question: 'Do you have or have you had cancer?' was carried out. We assessed correctness of the response in relation to cancer site, date of diagnosis, marital status, age and education. Smoking was chosen as a marker of health awareness. A total of 65,330 persons participated in the Nord-Trondelag Health Survey (HUNT-II), performed in 1995-1997. The database of HUNT-II was merged with the Cancer Registry of Norway (CRN), thus identifying each of the 2983 (4 percent) participants with an invasive cancer diagnosis. RESULTS: Excluding basal cell epithelioma, a total of 20 percent of the patients denied their prior cancer diagnosis. This group consisted mainly of men (54 percent) and those who were diagnosed as very young or as elderly. More smokers than non smokers were unaware of their prior malignancy (24 percent versus 20 percent). CONCLUSIONS: A 20 percent rate of patients who denied their former malignancy is surprisingly and unacceptably high. Disclosure of a cancer diagnosis should help the patient to develop increased health awareness. It should enable a person to report his or her former cancer diagnosis when necessary. PMID- 14747591 TI - A model to predict the results of changes in smoking behaviour on smoking prevalence. AB - BACKGROUND: Data are available on the prevalence of smoking states (never, current and ex). However, data on behaviour change rates (starting - never to current, quitting - current to ex and lapsing - ex to current) are not readily available and cannot be simply derived from or related to prevalence data. METHOD: A model was constructed to relate prevalence of smoking states to behaviour change rates. It was populated with prevalence of smoking status taken from the General Household Survey together with population structure, age- and sex-specific death rates, and birth rates for England and Wales. This model could be used to calculate past behaviour change given observed prevalence of smoking states or future prevalence of smoking given predicted rates of behaviour change. RESULTS: To fit data it was necessary to assume that as they age some ex smokers reclassify themselves as never smokers. In the age band 16-19 years about 9 percent of never smokers start smoking, and about 5 percent of current smokers quit. In the age band 20-24 years the corresponding figures for starting are about 4 percent in males and 2 percent in females, and for quitting about 2 percent in both. In older age bands the percentages starting are zero or less than zero (indicating reclassifying), and the percentage quitting rises with age. Net lapsing (shift from ex to current) occurred very infrequently and is quantitatively unimportant. If the current starting, quitting and lapsing rates are maintained the Smoking kills target will not be met. Future prevalence of smoking under different scenarios is examined. CONCLUSION: The model is useful in calculating the proportions changing smoking state from serial cross-sectional data on prevalence and for predicting future prevalence. PMID- 14747592 TI - Systematic review of the use and value of computer simulation modelling in population health and health care delivery. AB - BACKGROUND: The objective of the review was to evaluate the extent, quality and value of computer simulation modelling in population health and health care delivery. METHODS: A narrative systematic review was carried out of world literature from 1980 to 1999, searching Medline, INSPEC, Embase, HealthSTAR, Science Citation Index, CINAHL, MathSci, INFORMS Online and SIGLE databases, and researchers in the field were contacted. Papers were included if they contained a computer simulation model of individuals in a stochastic system and the topic or setting related to population health or health service delivery. RESULTS: A total of 182 papers met the inclusion criteria. Simulation modelling has been undertaken in a wide range of health care topic areas, including hospital scheduling and organization, communicable disease, screening, costs of illness and economic evaluation. However, the quality of published papers was variable and few reported on the outcomes of implementation of models, so that the value of modelling could not be assessed. CONCLUSION: Simulation modelling is a powerful method for modelling both small and large populations to inform policy makers in the provision of health care. It has been applied to a wide variety of health care problems. Although the number of modelling papers has grown substantially over recent years, further research is required to assess the value of modelling. PMID- 14747593 TI - Caring-related inequalities in psychological distress in Britain during the 1990s. AB - BACKGROUND: This paper examines recent trends in inequalities in psychological distress associated with the provision of unpaid care by those who look after frail older people and younger disabled adults and children. Caring activities intensified during the 1990s, associated with increasing amounts of time devoted to the more demanding types of care and to those relationships that typically make heavy demands on the carer. Heavy involvement in caregiving is often associated with symptoms of anxiety and distress, and the intensification of care may increase rates of distress in carers relative to that in non-carers. METHODS: A secondary analysis was carried out of data drawn from the first 10 waves of the British Household Panel Survey covering 1991-2000, based on around 9000 adults interviewed personally in successive waves. Symptoms of psychological distress, including anxiety and depression, were assessed using the 12-item General Health Questionnaire. RESULTS: Carers present higher rates of distress than noncarers and the health gap widens as the definition of caregiving focuses on those living with the person they care for, and those devoting 20 h or more per week to their caring activities. Differences in distress rates between carers and non-carers are greater for women than for men. There is no support for the hypothesis that inequalities in distress associated with caregiving have increased over time. CONCLUSION: There was no change during the 1990s in the extent of inequalities in psychological distress associated with caregiving in Britain. The need to maintain carers' emotional and mental health is as compelling as ever it was. PMID- 14747594 TI - Equity of access to tertiary hospitals in Wales: a travel time analysis. AB - BACKGROUND: The objective of the study was to investigate the implications for equity of geographical access for population subgroups arising from hypothetical scenarios of change in configuration of National Health Service tertiary hospital service provision located in Wales. METHODS: For each of three scenarios, the status quo and centralization of services to one of two locations, we used a travel time road length matrix in geographical information software to calculate the proportion of the population living within 30, 60, 90 and 120 min travel of each hospital site and the associated mean, median and 90th percentile travel times. We analysed data for the total resident population of Wales, for residents aged 75 or more years, for residents of the most deprived 10 per cent of enumeration districts, and for residents of rural areas. RESULTS: Centralization of services reduces geographical access for all population subgroups. Access varies between population subgroups, both between and within different scenarios of service configuration. A change in service configuration may improve access for one subgroup but reduce access for another. The interpretation may also vary according to whether the defined cut point for comparing access is based on short or long travel times. Measurements of absolute and relative access are sensitive to the assumed travel speeds. CONCLUSION: Access for the total population does not imply equity of access for subgroups of the population. Comparisons of access between scenarios are dependent on which measure of access is the indicator of choice. Results are sensitive to the road network travel speeds and further local validation may be necessary. This method can provide explicit information to health service planners on the effects on equity of access from a change in service configuration. PMID- 14747595 TI - Cement, cancers and clusters: an investigation of a claim of a local excess cancer risk related to a cement works. AB - BACKGROUND: We investigated claims by a campaigning group of a cancer cluster associated with a local cement works. METHODS: To investigate cancer rates in the town we defined the study area as the Census wards matching the geographical area code supplied to the campaigning group. Standard methods were applied to registered cases of cancer for the area for the years 1974-1989 to derive observed and expected numbers. The significance of the relative risk was assessed using the Poisson distribution. By selecting a different denominator population we attempted to reproduce the results of the campaign group. Cancer rates around the cement works were investigated for four cancer types plausibly associated with emissions, using cancer registrations for the years 1985-1994. Cases were mapped to 1981 Census ward boundaries, and the same statistical methods were used, but expected counts were also adjusted for deprivation. Rates were calculated for an inner 2 km zone and outer zone 2-5 km from the works. Relative risk was calculated and the ratio of risks was examined for evidence of increased risk closer to the works. RESULTS: Relative risks were not significantly elevated either in the town or around the cement works itself. We were able to reproduce the likely errors that resulted in the elevated relative risks for five cancer groups claimed by the campaigning group. CONCLUSIONS: We found no evidence of increased incidence of cancer around the cement works. Incorrect handling of cancer registration data can result in spurious cancer clusters and unnecessary public alarm. PMID- 14747596 TI - A survey of hepatitis C prevalence amongst the homeless community of Oxford. AB - Hepatitis C (HCV) is an emerging health concern across the world, with 170 million people chronically infected and at risk of liver cancer, cirrhosis or liver failure. There is no vaccination and so it is important to learn as much as possible about how to prevent future infection. Modes of transmission include intravenous drug use (IDU), blood products, tattooing and, to a lesser extent, sexual intercourse. Homelessness is a risk factor of HCV because of the environments and behaviours associated with homeless communities such as poor hygiene, poor nutrition and high levels of IDU. The aim of this project was to determine the prevalence of HCV and its risk factors amongst the homeless community of Oxford, which is the second largest in the country. Ninety-eight individuals of the Oxford homeless community were interviewed and tested for HCV. The results gave an estimated HCV prevalence of 26.5 percent. The major risk factors in this population were IDU (past and present), age (over 20 years old) and sharing the paraphernalia used by i.v. drug users (e.g. spoons, foil and filters). With the exception of age, these risk factors could all be targeted in an attempt to reduce this prevalence and combat the major public health concern that HCV poses to the homeless community of Oxford. PMID- 14747597 TI - Early warning and NHS Direct: a role in community surveillance? AB - BACKGROUND: NHS Direct is a nurse-led telephone help line that covers the whole of England and Wales. NHS Direct derived data are being used for community surveillance, the purpose of which is to detect a local or national increase in symptoms reported by callers. The system has the potential to identify an increase in symptoms reported by callers about people in the prodromal stages of illness caused by the deliberate release of a biological or chemical agent. There are no other community surveillance projects existing on a national scale that utilize electronic daily data. METHODS: We describe the surveillance system and calls to NHS Direct between December 2001 and July 2002. Confidence limits have been constructed for 10 key algorithms at each site and control charts devised for five of these algorithms at sites covering the key urban areas. RESULTS: Daily reporting has been achieved from NHS Direct sites in England and Wales. High levels of activity in specific algorithms at both national and regional levels have been detected. A sustained national increase in calls about fever occurred in January 2002. CONCLUSION: Although the project is still at an early stage, daily analysis of NHS Direct data has the potential to detect symptoms in the community that could be related to deliberate releases of chemical or biological agents or to outbreaks of disease. For this surveillance to act as an 'early warning' of illness resulting from a microbiological or chemical cause, the NHS Direct surveillance needs to be fully integrated into an appropriate public health response (which may require diagnostic samples to be taken from callers). PMID- 14747598 TI - Health effects of a sulphur dioxide air pollution episode. AB - BACKGROUND: A sulphur dioxide (SO(2)) episode occurred in the United Kingdom in 1998. The worst affected area was the city of Nottingham. METHODS: Emergency hospital admissions in Nottingham in the episode week were compared with those in the previous week. RESULTS: A statistically significant increase in admissions for all respiratory diseases occurred in the episode week (odds ratio (OR) = 1.40, 95 per cent confidence interval (CI) = 1.00-1.94). Ten of the 25 excess admissions were for asthma, although the excess for asthma alone was not statistically significant (OR = 1.90, 95 per cent CI = 0.87-4.15). CONCLUSIONS: The excess admissions for respiratory diseases could have been caused by exposure to SO(2), to other pollutants present in increased concentrations during the pollution episode, or by seasonal variations in the frequency of asthma symptoms, or prevailing weather conditions. This study shows how simple analyses of routinely collected health data can be used to assess public health impacts of pollution episodes. PMID- 14747599 TI - Patterns of adult and old-age mortality in rural Burkina Faso. AB - Based on a demographic surveillance population from 39 villages in rural Burkina Faso, we describe mortality patterns in adults (15-59 years) and older people (> or = 60 years), and discuss seasonal trends in mortality. During the study period 1993-1998, 589 deaths in adults and 593 deaths in older people were recorded from an average adult and older people population of 13 550. The crude all-cause mortality rate per 1000 for adults was 7.3 (95 per cent confidence interval (CI) 6.7-7.8) and for older people 55.8 (95 percent CI 51.3-60.3). The probability of dying before age 60 after reaching age 15 was 34 percent for males and 32 percent for females. Malaria and diarrhoea, recorded through verbal autopsy, accounted for 21 percent of total deaths in adults and 22 per cent in older people. A seasonal trend in mortality for older people with a peak in February was identified. The study shows that malaria is an important cause of death in adulthood. PMID- 14747600 TI - Prophylactic aspirin use in the adult general population. AB - BACKGROUND: The aim of the study was to establish the prevalence and patterns of aspirin use in people with vascular problems. METHODS: A cross-sectional population survey was carried out on a stratified random sample of 10,000 adults aged 35 and over in North Staffordshire. RESULTS: A total of 6322 adults replied to the questionnaire (adjusted response 67 percent). The prevalence of vascular problems was 12.9 percent, and 67.6 per cent of respondents were using aspirin. The main association with aspirin use was previous advice about aspirin: adults who recalled being given advice were more likely to be using aspirin. Increasing age, disease severity and level of deprivation were also associated with increased aspirin use. Of adults without vascular problems, 7.1 percent also reported using aspirin regularly. CONCLUSIONS: There is still potential to increase aspirin use in those with vascular problems. The extent and quality of health care professionals' advice may be an important area to target. The reasons why some people without vascular problems take regular aspirin is an area for further investigation. PMID- 14747601 TI - Communicable disease and health protection quarterly review: April to June 2003. From the Health Protection Agency Communicable Disease Surveillance Centre. PMID- 14747603 TI - Change in adult health following medical priority rehousing. PMID- 14747604 TI - Change in adult health following medical priority rehousing. PMID- 14747605 TI - Contact tracing and population screening for tuberculosis - who should be assessed? PMID- 14747609 TI - SSR240612 [(2R)-2-[((3R)-3-(1,3-benzodioxol-5-yl)-3-[[(6-methoxy-2 naphthyl)sulfonyl]amino]propanoyl)amino]-3-(4-[[2R,6S)-2,6 dimethylpiperidinyl]methyl]phenyl)-N-isopropyl-N-methylpropanamide hydrochloride], a new nonpeptide antagonist of the bradykinin B1 receptor: biochemical and pharmacological characterization. AB - The biochemical and pharmacological properties of a novel non-peptide antagonist of the bradykinin (BK) B(1) receptor, SSR240612 [(2R)-2-[((3R)-3-(1,3-benzodioxol 5-yl)-3-[[(6-methoxy-2-naphthyl)sulfonyl]amino]propanoyl)amino]-3-(4-[[2R,6S)-2,6 dimethylpiperidinyl]methyl]phenyl)-N-isopropyl-N-methylpropanamide hydrochloride] were evaluated. SSR240612 inhibited the binding of [(3)H]Lys(0)-des-Arg(9)-BK to the B(1) receptor in human fibroblast MRC5 and to recombinant human B(1) receptor expressed in human embryonic kidney cells with inhibition constants (K(i)) of 0.48 and 0.73 nM, respectively. The compound selectivity for B(1) versus B(2) receptors was in the range of 500- to 1000-fold. SSR240612 inhibited Lys(0) desAr(9)-BK (10 nM)-induced inositol monophosphate formation in human fibroblast MRC5, with an IC(50) of 1.9 nM. It also antagonized des-Arg(9)-BK-induced contractions of isolated rabbit aorta and mesenteric plexus of rat ileum with a pA(2) of 8.9 and 9.4, respectively. Antagonistic properties of SSR240612 were also demonstrated in vivo. SSR240612 inhibited des-Arg(9)-BK-induced paw edema in mice (3 and 10 mg/kg p.o. and 0.3 and 1 mg/kg i.p.). Moreover, SSR240612 reduced capsaicin-induced ear edema in mice (0.3, 3 and 30 mg/kg p.o.) and tissue destruction and neutrophil accumulation in the rat intestine following splanchnic artery occlusion/reperfusion (0.3 mg/kg i.v.). The compound also inhibited thermal hyperalgesia induced by UV irradiation (1 and 3 mg/kg p.o.) and the late phase of nociceptive response to formalin in rats (10 and 30 mg/kg p.o.). Finally, SSR240612 (20 and 30 mg/kg p.o.) prevented neuropathic thermal pain induced by sciatic nerve constriction in the rat. In conclusion, SSR240612 is a new, potent, and orally active specific non-peptide bradykinin B(1) receptor antagonist. PMID- 14747610 TI - Adenovirus-mediated delivery and expression of a cAMP-dependent protein kinase inhibitor gene to BEAS-2B epithelial cells abolishes the anti-inflammatory effects of rolipram, salbutamol, and prostaglandin E2: a comparison with H-89. AB - cAMP-elevating drugs are thought to mediate their biological effects by activating the cAMP/cAMP-dependent protein kinase (PKA) cascade. However, this hypothesis is difficult to confirm due to a lack of selective inhibitors. Here, we have probed the role of PKA in mediating inhibitory effects of several cAMP elevating drugs in BEAS-2B epithelial cells using an adenovirus vector encoding a PKA inhibitor protein (PKIalpha) and have compared it to H-89, a commonly used small molecule PKA inhibitor. Initial studies established efficient gene transfer and confirmed functionality of PKIalpha 48 h after virus infection. All cAMP elevating drugs tested promoted the phosphorylation of cAMP response element binding protein (CREB), activated a cAMP response element (CRE)-driven luciferase reporter gene, and suppressed both granulocyte/macrophage colony-stimulating factor (GM-CSF) generation and [(3)H]arachidonic acid (AA) release in response to interleukin-1beta and monocyte chemotactic protein (MCP)-1, respectively. These effects were abolished by PKIalpha. In contrast, H-89 behaved unpredictably under the same conditions. Thus, although CREB phosphorylation evoked by a range of cAMP-elevating drugs was abolished by H-89, neither activation of the CRE dependent luciferase reporter gene construct nor the inhibition of GM-CSF generation was inhibited. Paradoxically, H-89 antagonized MCP-1-induced [(3)H]AA release and enhanced the inhibitory effect of submaximal concentrations of rolipram and 8-bromo-cAMP. We suggest that expression of PKIalpha in susceptible cells provides a simple and unambiguous way to assess the role of PKA in cAMP signaling and to probe the mechanism of action of other drugs and cAMP-dependent responses where the participation of PKA is equivocal. Furthermore, these data suggest that H-89 is not a selective inhibitor of PKA and should be avoided. PMID- 14747611 TI - Risk of ventricular proarrhythmia with selective opening of the myocardial sarcolemmal versus mitochondrial ATP-gated potassium channel. AB - Myocardial ATP-gated potassium channels (K-ATPs) are critical in the intracellular signaling cascade resulting in ischemic preconditioning (IP). Mitochondrial K-ATP channels seem to be responsible for IP, whereas the functions of K-ATP channels in the sarcolemmal membrane are less well understood. The proarrhythmic potential of specific versus nonspecific opening of K-ATP channels has not been investigated. In this study, Langendorff-perfused rabbit hearts were exposed to either pinacidil (1.25 microM), a nonselective K-ATP channel agonist, or selective mitochondrial or sarcolemmal K-ATP channel agonists or antagonists. The hearts were then subjected to 12 min of hypoxic perfusion and 40 min of reoxygenation. Hearts were monitored for the induction of ventricular fibrillation (VF). No heart subjected to hypoxia-reoxygenation without drug treatment developed VF (0 of 5). Pinacidil pretreatment induced VF (12 of 14; p = 0.004 versus control). Pinacidil's effect was blocked by HMR-1098 (1-[5-[2-(5 chloro-o-anisamide)ethyl]-2-methoxyphenyl]sulfonyl]-3-methylthiourea) (1 microM), a selective sarcolemmal K-ATP channel antagonist (1 of 7; p = 0.007 versus pinacidil; N.S. versus control). Hearts pretreated with 5-hydroxydecanoate (5-HD) (100 microM), a putatively selective mitochondrial K-ATP channel blocker developed VF in one of eight trials (N.S. versus control). 5-HD did not alter the effects of pinacidil (6 of 8; p < 0.05 versus control; N.S. versus pinacidil alone). Selective mitochondrial K-ATP channel activation with [(3R)-trans-4-((4 chlorophenyl)-N-(1H-imidazol-2-ylmethyl)dimethyl-2H-1-benzopyran-6-carbonitril monohydrochloride] (BMS-191095) (6 microM) resulted in zero of five hearts developing VF (N.S. versus control). Our data suggest that selective opening of the sarcolemmal K-ATP channel during hypoxia-reoxygenation induced VF, whereas opening of the mitochondrial channel was not associated with VF. The findings suggest that caution should be exercised when developing compounds aimed at inducing IP, and nonspecific opening of the K-ATP channel should be avoided. PMID- 14747612 TI - Cardioprotection of interleukin-2 is mediated via kappa-opioid receptors. AB - We examined whether interleukin-2 (IL-2) protects the myocardium against injury induced by ischemia and reperfusion via the kappa-opioid receptor (OR). The cardioprotective effect of IL-2 was evaluated by measuring infarct size and lactate dehydrogenase (LDH) release in response to ischemia and reperfusion in the isolated rat heart. IL-2 at an optimal dose of 50 U/ml mimicked the effect of ischemic preconditioning by reducing infarct size and LDH release. The infarct and LDH-reducing effects of IL-2 were blocked by nor-binaltorphimine (5 microM), a kappa-OR antagonist, but not naltrindole (5 microM), a delta-OR antagonist known to block the action of its stimulation. Moreover, blockade of the mitochondrial ATP-sensitive potassium (mito-K(ATP)) channel with a selective antagonist, 5-hydroxydecanoate (100 microM), or a nonselective antagonist of K(ATP) channels, glybenclamide (100 microM), or blockade of protein kinase C (PKC) with its inhibitors chelerythrine (5 microM) or GF 109203X (10 microM) [3 [1-[3-(dimethylaminopropyl]-1H-indol-3-yl]-4-(1H-indol-3-yl)-1H-pyrrole-2,5-dione monohydrochloride] abolished the protective effect of IL-2. Administration of free radical scavengers N-acetylcysteine (4 mM) or N-(2-mercaptopropionyl) glycine (1 mM) also abolished the protective effects of IL-2 and U50,488H [(trans)-3,4-dichloro-N-methyl-N-[2-(1-pyrrolidinyl) cyclohexyl]benzeneacetamide], a selective kappa-OR agonist. This study provides the first evidence that IL-2 confers cardioprotection against injury induced by ischemia/reperfusion. The effect of IL-2 is mediated via kappa-OR as evidenced by kappa-OR antagonism and similar signaling mechanisms, mito-K(ATP), PKC, and reactive oxygen species involved in the cardioprotective effects of both IL-2 and kappa-OR stimulation. PMID- 14747613 TI - A novel selective allosteric modulator potentiates the activity of native metabotropic glutamate receptor subtype 5 in rat forebrain. AB - We found that N-[4-chloro-2-[(1,3-dioxo-1,3-dihydro-2H-isoindol-2 yl)methyl]phenyl]-2-hydroxybenzamide (CPPHA), is a potent and selective positive allosteric modulator of the metabotropic glutamate receptor subtype 5 (mGluR5). CPPHA alone had no agonist activity and acted as a selective positive allosteric modulator of human and rat mGluR5. CPPHA potentiated threshold responses to glutamate in fluorometric Ca(2+) assays 7- to 8-fold with EC(50) values in the 400 to 800 nM range, and at 10 microM shifted mGluR5 agonist concentration response curves to glutamate, quisqualate, and (R,S)-3,5-dihydroxyphenylglycine (DHPG) 4- to 7-fold to the left. The only effect of CPPHA on other mGluRs was weak inhibition of mGluR4 and 8. Neither CPPHA nor the previously described 3,3' difluorobenzaldazine (DFB) affected [(3)H]quisqualate binding to mGluR5, but although DFB partially competed for [(3)H]3-methoxy-5-(2 pyridinylethynyl)pyridine binding, CPPHA had no effect on the binding of this 2 methyl-6-(phenylethynyl)-pyridine analog to mGluR5. Although the binding sites for the two classes of allosteric modulators seem to be different, these different allosteric sites can modulate functionally and mechanistically similar allosteric effects. In electrophysiological studies of brain slice preparations, it had been previously shown that activation of mGluR5 receptors by agonists increased N-methyl-D-aspartate (NMDA) receptor currents in the CA1 region of hippocampal slices. We found that CPPHA (10 microM) potentiated NMDA receptor currents in hippocampal slices induced by threshold levels of DHPG, whereas having no effect on these currents by itself. Similarly, 10 microM CPPHA also potentiated mGluR5-mediated DHPG-induced depolarization of rat subthalamic nucleus neurons. These results demonstrate that allosteric potentiation of mGluR5 increases the effect of threshold agonist concentrations in native systems. PMID- 14747614 TI - Tumor necrosis factor-alpha-induced cytokine-induced neutrophil chemoattractant-1 (CINC-1) production by rat gastric epithelial cells: role of reactive oxygen species and nuclear factor-kappaB. AB - Rat cytokine-induced neutrophil chemoattractant-1 (CINC-1), a counterpart of the human growth-regulated oncogene product (GRO), has been suggested to participate in neutrophil recruitment in an experimental model of gastritis in rat. However, the mechanism(s) involved in regulation of CINC-1 production by the gastric mucosa remains unclear. The aim of this study was to investigate the mechanism(s) of CINC-1 production by rat gastric mucosa in vitro. All experiments were performed using rat normal gastric mucosal cell line (RGM-1). RGM-1s were stimulated with tumor necrosis factor (TNF)-alpha, and CINC-1 mRNA levels (reverse transcription-polymerase chain reaction) and protein secretion (enzyme linked immunosorbent assay) were assessed. The production of reactive oxygen species (ROS) and nuclear factor (NF)-kappaB activation (translocation to the nuclei) in response to TNF-alpha stimulation was evaluated using fluorescence microscopy in the presence or absence of the inhibitors of mitochondrial electron flow and NF-kappaB activation. Stimulation of RGM-1 cells with TNF-alpha resulted in an increase in intracellular oxidative stress, NF-kappaB translocation to the nuclei, and up-regulation of CINC-1 mRNA and protein, which was prevented by interfering with mitochondria-dependent ROS production and NF-kappaB activation. Taken together, these findings indicate that CINC-1, a counterpart of the human GRO, production by rat gastric epithelial cells in response to TNF-alpha stimulation is an oxidant stress-mediated and NF-kappaB-dependent event. PMID- 14747615 TI - Alterations in vesicular dopamine uptake contribute to tolerance to the neurotoxic effects of methamphetamine. AB - Previous studies demonstrated that tolerance to the long-term neurotoxic effects of methamphetamine on dopamine neurons could be induced by pretreating with multiple injections of escalating doses of methamphetamine. The mechanism(s) underlying this tolerance phenomenon is unknown. Some recent studies suggested that aberrant vesicular monoamine transporter-2 (VMAT-2) and dopamine transporter function contribute to neurotoxic effects of methamphetamine. Hence, the purpose of this study was to explore the role of the VMAT-2 and dopamine transporter in the induction of tolerance to the longterm persistent dopaminergic deficits caused by methamphetamine. A second purpose was to investigate the potential role of hyperthermia and alterations in brain methamphetamine distribution in this tolerance. Results revealed that the methamphetamine pretreatment regimen attenuated both the acute methamphetamine-induced decrease in VMAT-2 function 2 h after the methamphetamine challenge administration and its resulting persistent dopamine deficits without attenuating the acute methamphetamine-induced decreases in dopamine transporter uptake. Furthermore, pretreatment with methamphetamine prior to a high-dose methamphetamine challenge administration also attenuated the acute methamphetamine-induced redistribution of VMAT-2 immunoreactivity within the nerve terminal. This protection was not due to alterations in concentration of methamphetamine in the brain because both the methamphetamine- and saline pretreated rats had similar amounts of methamphetamine and amphetamine at 30 min to 2 h after the last methamphetamine challenge injection. In summary, these data are the first to demonstrate an association between the prevention of acute alterations in vesicular dopamine uptake and the development of tolerance to the neurotoxic effects of methamphetamine. PMID- 14747616 TI - Oxidation of protein tyrosine phosphatases as a pharmaceutical mechanism of action: a study using 4-hydroxy-3,3-dimethyl-2H-benzo[g]indole-2,5(3H)-dione. AB - Growth factor and insulin signal transduction comprise series of protein kinases and protein phosphatases whose combined activities serve to propagate the growth factor signal in a regulated fashion. It was shown previously that such signaling cascades generate hydrogen peroxide inside cells. Recent work has implied that one function of this might be to enhance the feed-forward signal through the reversible oxidation and inhibition of protein tyrosine phosphatases (PTPs). We identified compound 4-hydroxy-3,3-dimethyl-2H-benzo[g]indole-2,5(3H)-dione (BVT.948) as an agent that is able to inhibit PTP activity in vitro noncompetitively, a mechanism involving oxidation of the catalytic cysteine residue. We investigated the pharmaceutical utility of this compound by examining its effects in a series of in vitro cellular and in vivo assays. Results showed that BVT.948 was able to enhance insulin signaling in cells, although it did not increase tyrosine phosphorylation globally. Furthermore, the compound was active in vivo, enhancing insulin tolerance tests in ob/ob mice, therefore apparently enhancing insulin sensitivity. BVT.948 was able to inhibit several other PTPs tested and also was efficient at inhibiting several cytochrome P450 (P450) isoforms in vitro. The data suggest that inhibitors of PTPs that display noncompetitive kinetics must be viewed with caution because they may oxidize the enzyme irreversibly. Furthermore, although such compounds display interesting biological effects in vitro and in vivo, their general pharmaceutical utility may be limited due to undesired effects on P450 enzymes. PMID- 14747617 TI - Immune cell regulation and cardiovascular effects of sphingosine 1-phosphate receptor agonists in rodents are mediated via distinct receptor subtypes. AB - Sphingosine 1-phosphate (S1P) is a bioactive lysolipid with pleiotropic functions mediated through a family of G protein-coupled receptors, S1P(1,2,3,4,5). Physiological effects of S1P receptor agonists include regulation of cardiovascular function and immunosuppression via redistribution of lymphocytes from blood to secondary lymphoid organs. The phosphorylated metabolite of the immunosuppressant agent FTY720 (2-amino-2-(2-[4-octylphenyl]ethyl)-1,3 propanediol) and other phosphonate analogs with differential receptor selectivity were investigated. No significant species differences in compound potency or rank order of activity on receptors cloned from human, murine, and rat sources were observed. All synthetic analogs were high-affinity agonists on S1P(1), with IC(50) values for ligand binding between 0.3 and 14 nM. The correlation between S1P(1) receptor activation and the ED(50) for lymphocyte reduction was highly significant (p < 0.001) and lower for the other receptors. In contrast to S1P(1) mediated effects on lymphocyte recirculation, three lines of evidence link S1P(3) receptor activity with acute toxicity and cardiovascular regulation: compound potency on S1P(3) correlated with toxicity and bradycardia; the shift in potency of phosphorylated-FTY720 for inducing lymphopenia versus bradycardia and hypertension was consistent with affinity for S1P(1) relative to S1P(3); and toxicity, bradycardia, and hypertension were absent in S1P(3)(-/-) mice. Blood pressure effects of agonists in anesthetized rats were complex, whereas hypertension was the predominant effect in conscious rats and mice. Immunolocalization of S1P(3) in rodent heart revealed abundant expression on myocytes and perivascular smooth muscle cells consistent with regulation of bradycardia and hypertension, whereas S1P(1) expression was restricted to the vascular endothelium. PMID- 14747619 TI - Politics and health. PMID- 14747618 TI - Expression of Fcgamma and complement receptors on peripheral blood monocytes in systemic lupus erythematosus and rheumatoid arthritis. AB - OBJECTIVES: Fcagamma and complement receptors play an important role in the interaction between immune complexes (IC) and monocytes/macrophages. Recent work has demonstrated that their relative expression on these cells may be modified by cytokines, including TNF-alpha and IL-4. Furthermore, cytokines may alter the expression of adhesion molecules such as ICAM-1. However, little data exist on the in vivo expression of specific Fcgamma and complement receptors in systemic lupus erythematosus (SLE) or rheumatoid arthritis (RA), two diseases in which IC are important in pathogenesis. METHODS: Venous blood was obtained from 30 patients with SLE, 25 with RA and 25 healthy controls. Monocyte phenotype was determined by flow cytometric analysis of whole blood samples, with selective gating using forward and side scatter signals. Surface expression of Fcgamma receptors RI (CD64), RII (CD32) and RIII (CD16), complement receptors CR1 (CD35) and CR3 (CD11b/CD18), and adhesion molecules ICAM-1 (CD54) and CD11a (LFA-1) was determined. The effects of disease activity and corticosteroid therapy on the expression of these molecules were also examined. RESULTS: The expression of FcgammaRII was reduced on monocytes from patients with SLE compared with healthy controls and patients with RA (P = 0.002). This did not correlate with disease activity using conventional indices [SLEDAI (SLE disease activity index), C3/C4 levels and anti-double-stranded DNA antibody titres], and was independent of prednisolone therapy. There was no significant difference in FcgammaRI or RIII expression on SLE monocytes compared with healthy controls. In contrast, the expression of FcgammaRIII was increased on RA monocytes (P = 0.01), this being highest in patients with active disease. The proportion of FcgammaRIII-positive monocytes was also increased in RA, and prednisolone therapy was associated with a lower proportion of FcgammaRIII-positive cells. An increase in CR3 expression was seen on RA monocytes (P = 0.002), whilst CR1 was increased on monocytes from patients with active SLE or active RA. ICAM-1 expression was reduced on monocytes from patients with SLE (P = 0.002), although high-dose prednisolone therapy was associated with the lowest level of surface ICAM-1 on monocytes. CONCLUSIONS: Peripheral blood monocytes from patients with SLE or RA display significantly altered phenotypes compared with those from healthy controls. The observed reduction in SLE of FcgammaRII may represent a mechanism by which monocytes are protected from IC-mediated activation. Prednisolone therapy and disease activity had little effect on phagocytic receptor expression. The observed changes may reflect the different cytokine profiles seen in SLE and RA. PMID- 14747620 TI - Acute and long-term management of patients with vertebral fractures. AB - Symptomatic vertebral fractures are associated with significant morbidity, excess mortality and health and social service expenditure. Up to 20% of patients with an incident vertebral fracture experience a further vertebral fracture within one year. It is therefore important that vertebral fractures are detected early, and treatment considered as soon as possible. Only a third of vertebral fractures come to medical attention, where they typically present with acute back pain, but other presentations include loss of height and increasing kyphosis. Spine X-rays should then be performed to confirm the diagnosis and exclude other pathology. Bone density measurements are not essential before starting treatment for osteoporosis in patients with low-trauma vertebral fractures, but may be useful to confirm osteoporosis when there is uncertainty about previous trauma. They may also aid in selecting the most appropriate therapy and monitoring response to treatment. Up to 30% of women and 55% of men with symptomatic vertebral crush fractures have underlying secondary osteoporosis, where treatment may lead to large increases in bone density. These conditions should therefore be sought by medical history, physical examination and appropriate investigations. The management of patients with acute vertebral fractures should include measures to reduce pain and improve mobility, as well as starting treatment for osteoporosis. Treatments have now been shown in randomized controlled trials to improve bone density and reduce the incidence of vertebral and non-vertebral fractures in patients with osteoporosis. Choice of treatment will depend on the underlying causes of bone loss, efficacy in any particular situation, cost, patient preference and the potential non-skeletal advantages and disadvantages. PMID- 14747621 TI - Secondary contamination in organophosphate poisoning: analysis of an incident. AB - BACKGROUND: Acute organophosphate poisoning is rare in the UK, and the risks to attending staff are seldom appreciated. STUDY DESIGN: Report of an incident. RESULTS: In May 2001, a 45-year-old man attempted suicide by drinking organophosphate insecticide, and was brought to an urban general hospital in a collapsed state. Twenty-five hospital workers and paramedics sought medical advice after coming into contact with the poisoned patient, of whom ten complained of symptoms related to toxin exposure. Provision of emergency services by the hospital was compromised, and the emergency department was closed until the area was decontaminated and staffing levels could be restored. DISCUSSION: Ingestion of OP compounds can present a significant risk to health professionals as well as patients. Problems with the management of this patient included late recognition of the need for decontamination, large numbers of non-essential staff coming into contact with the patient, and the difficulty of carrying out medical procedures while wearing protective equipment. Decontamination should always be considered early, and the possibility of an ingested poison being vomited and causing a chemical spill should not be overlooked. PMID- 14747622 TI - One-year follow-up of heart failure patients after their first admission. AB - BACKGROUND: Outcomes related to heart failure remain relatively poor. AIM: To examine the clinical course of patients for one year after their first admission because of heart failure, including prognosis, mortality, and rehospitalization. DESIGN: Prospective observational study. METHODS: Of 121 patients hospitalized over 6 months for decompensation of previously unknown heart failure, we excluded those with a possible previous diagnosis of heart failure (n = 5), who suffered from another serious disease with a poor prognosis (n = 6), died during the index hospitalization (n = 5), refused to participate (n = 4) or were lost to follow-up (n = 6). Mortality and readmissions were identified by prospective follow-up of all patients. RESULTS: Of the 98 patients evaluated, half (49) were women. Mean +/- SD age was 75.2 +/- 12 years. The 1-year case-fatality rate after the first admission was 24%; 19% of the deaths were heart failure-related, with progressive pump failure the predominant cause (14% of the total). Age was the only factor associated with increased mortality (p < 0.007). Of the 74 survivors, 32% experienced at least one hospital readmission during follow-up. DISCUSSION: The prognosis of unselected new cases of heart failure after their first hospitalization remains relatively poor, despite recent advances in pharmacological therapy and medical care. PMID- 14747623 TI - Hypothetical economic analysis of screening for left ventricular hypertrophy in high-risk normotensive populations. AB - BACKGROUND: Left ventricular hypertrophy (LVH) measured by echocardiography is a powerful independent marker of increased cardiovascular risk. The prevalence of echocardiographic LVH in patients with high cardiovascular risk appears to be high, even in patients currently considered normotensive. AIM: To ascertain the likely costs of screening for and treating echocardiographic LVH in normotensive patients at high risk of cardiovascular events. DESIGN: Hypothetical economic analysis. METHODS: Cost analyses were based on known costs of echocardiography, costs of selected cardiovascular medications and prevalence of normotensive LVH in at-risk populations, combined with treatment effect data from studies of hypertensive patients with echocardiographic LVH. RESULTS: Screening costs per case for echocardiographic LVH are likely to be low, because of the high prevalence of the condition and the low unit cost of echocardiography. Treatment costs are likely to be comparable to those currently deemed acceptable in treating high-risk cardiovascular populations, e.g. the HOPE study population. DISCUSSION: The costs of screening for and treating LVH in normotensive patients at risk of cardiovascular events do not appear to be prohibitively high. Trials of screening and treatment for normotensive LVH seem therefore to be warranted. PMID- 14747624 TI - Bone mineral status in immigrant Indo-Asian women. AB - BACKGROUND: Indo-Asian immigrants are known to be at high risk of metabolic bone disease, but the prevalence of osteoporosis in this population is unknown. AIM: To compare the bone mineral at the lumbar spine and femoral neck of Indo-Asian immigrant women with that of age-matched Caucasian women. DESIGN: Retrospective analysis. METHODS: Women of Indo-Asian origin referred for bone density scans in the last five years were identified. The skeletal status of each was compared with an age-matched Caucasian control for bone mineral content (BMC), bone mineral density (BMD) and bone mineral apparent density (BMAD) at the lumbar spine and femoral neck, and hip axis length was measured. RESULTS: At the lumbar spine, Indo-Asians had a significantly lower BMD than Caucasians (0.834 vs. 0.913, p = 0.008), but there was no significant difference when BMAD values were calculated (0.123 vs. 0.122). At the femoral neck, there was no difference in BMD (0.728 vs. 0.712, p = 0.5), and BMAD values were significantly higher among Indo Asians than Caucasians (0.393 vs. 0.319, p = 0.022). Hip axis length was significantly shorter among Indo-Asian women (10.3 vs. 10.7, p = 0.009). DISCUSSION: Although Indo-Asian women appear to have lower spinal BMD than Caucasians, these differences disappear when BMAD values are calculated. While BMD is an areal density, not taking into account the 'depth' of the bone, BMAD is an estimation of volumetric density. Hence lower BMD values in Asians may be a size-related artefact. Longitudinal studies may be required to evaluate the use of BMD as a marker for fracture risk in this population. PMID- 14747625 TI - The patient with a fragility fracture: an evolving role for the orthopaedic surgeon. AB - Osteoporosis can now be diagnosed readily, and treatments that increase bone mineral density and decrease fracture risk, even after fragility fracture has occurred, are now available. Clinical guidelines for management of osteoporosis unanimously recognize that fracture risk is highest among those who have already sustained a fracture, and encourage prompt evaluation and treatment of these individuals. Despite these guidelines, most women who experience fragility fractures remain untreated (for osteoporosis) by any of the physicians involved in their care. Barriers to diagnosis and treatment have been identified, including uncertainty about the responsibility for such management. The orthopaedic surgeon has a unique opportunity to initiate definitive osteoporosis evaluation and treatment in patients who present with fractures, and recent guidelines support the evolution of the role of the orthopaedist in this direction. PMID- 14747626 TI - Flatulence and carpopedal spasm: more than social embarrassment. PMID- 14747627 TI - Familial juvenile hyperuricaemic nephropathy. PMID- 14747628 TI - Hearing the sermons in stones. PMID- 14747629 TI - Cultural nepotism. PMID- 14747631 TI - Cerebrovascular reactivity in major depression: a pilot study. AB - OBJECTIVE: There are a growing number of reports that depression may increase the risk of stroke. Little is known, however, about the pathophysiologic mechanisms underlying this association. Cerebrovascular reactivity (CVR) reflects the compensatory dilatory capacity of cerebral arterioles to a dilatory stimulus and is an important mechanism to provide constant cerebral blood flow. We hypothesized that CVR is reduced in patients with major depression, thus contributing to the association between depression and stroke. METHODS: We assessed CVR in 33 patients with unipolar depression and 26 healthy controls by calculating the increase in cerebral blood flow velocity after stimulation with acetazolamide. Blood flow velocities were measured by transcranial Doppler ultrasound. RESULTS: Cerebrovascular reactivity was significantly reduced in depressed patients. Smoking was also associated with a significant reduction in CVR, whereas age and gender had no significant influence. CONCLUSIONS: Cerebrovascular reactivity appears to be impaired in major depression. Further studies should clarify the mechanisms leading to this reduced CVR. PMID- 14747632 TI - Depressive symptoms are related to higher ambulatory blood pressure in people with a family history of hypertension. AB - OBJECTIVE: We investigated whether parental history of hypertension (FH+) enhances the impact of depressed mood, indexed by Beck Depression Inventory (BDI), on ambulatory blood pressure (ABP). METHODS: Twenty-four-hour ABP, urinary norepinephrine (NE), and cortisol were obtained in 314 unmedicated normotensive and hypertensive men and women (age 18-64 years) who also completed the BDI. RESULTS: Subjects with a positive family history of hypertension (N = 177) exhibited significantly greater mean body mass index (BMI) and ABP compared with subjects without (N = 137). Importantly, when covarying for age, BMI, gender, and race, linear regressions revealed significant FH by BDI interactions. Higher BDI scores were significantly associated with higher 24-hour ABP in FH+ subjects, but not in FH- participants. Relationships were significantly stronger in those with two hypertensive parents vs. those with one vs. those with no hypertensive parents. Increases in BDI scores were significantly related to greater heart rate (HR) and 24-hour urinary NE in both FH+ and FH- groups, although no evidence of a mediational role for NE in the effect of BDI score on blood pressure (BP) or HR was seen. CONCLUSIONS: These findings suggest that depressed mood may be reliably associated with higher BP only among those with an underlying susceptibility to HTN. PMID- 14747633 TI - Impact of pain on depression treatment response in primary care. AB - OBJECTIVE: Pain commonly coexists with depression, but its impact on treatment outcomes has not been well studied. Therefore, we prospectively evaluated the impact of comorbid pain on depression treatment response and health-related quality of life. METHODS: We analyzed data from the ARTIST study, a randomized controlled trial with naturalistic follow-up conducted in 37 primary care clinics. Participants were 573 clinically depressed patients randomized to one of three selective serotonin reuptake inhibitor (SSRI) antidepressants: fluoxetine, paroxetine, or sertraline. Depression as assessed by the Symptom Checklist-20 (SCL-20) was the primary outcome. Secondary outcomes included pain and health related quality of life. RESULTS: Pain was reported by more than two thirds of depressed patients at baseline, with the severity of pain mild in 25% of patients, moderate in 30%, and severe in 14%. After 3 months of antidepressant therapy, 24% of patients had a poor depression treatment response (ie, SCL-20 >1.3). Multivariate odds ratios for poor treatment response were 1.5 (95% confidence interval, 0.8-3.2) for mild pain, 2.0 (1.1-4.0) for moderate pain, and 4.1 (1.9-8.8) for severe pain compared with those without pain. Increasing pain severity also had an adverse impact on outcomes in multiple domains of health related quality of life. CONCLUSIONS: Pain is present in two thirds of depressed primary care patients begun on antidepressant therapy, and the severity of pain is a strong predictor of poor depression and health-related quality of life outcomes at 3 months. Better recognition, assessment, and treatment of comorbid pain may enhance outcomes of depression therapy. PMID- 14747634 TI - Hyperleptinemia in subjects with persistent partial posttraumatic stress disorder after a major earthquake. AB - OBJECTIVE: Leptin, a peptide hormone derived from adipose tissue, regulates food intake and controls weight. Serum leptin levels may be elevated in critically ill patients and in cases of physical stress. Our aim was to examine the relationship between postdisaster psychiatric symptoms and serum leptin levels. METHODS: We recruited 92 subjects who visited emergency medical stations immediately after the giant earthquake seized middle Taiwan on September 21, 1999. Of these cases, 43 met the criteria for subsyndromal acute stress disorder. Eighteen months later, we measured serum leptin levels and performed in-depth psychiatric assessments using the posttraumatic stress disorder subset in the Mini International Neuropsychiatric Instrument and the Brief Symptom Rating Scale short form. RESULTS: After we adjusted for age, gender, and body mass index, serum leptin levels were significantly higher in the persistent subsyndromal group (N = 15; estimated marginal mean, 9.641; 95% confidence interval, 7.673 11.610) than in others (estimated marginal means, 4.775-6.368). A hyperaroused state predicted a higher adjusted leptin level. On the Brief Symptom Rating Scale, general severity index, paranoid tendency, anxiety symptoms, and depressive symptoms were positively correlated with adjusted leptin levels. CONCLUSIONS: We found a direct relationship between stress-related psychopathologic symptoms and serum leptin levels. Subjects with persistent subsyndromal posttraumatic stress disorder and hyperarousal had significantly higher serum leptin levels. Therefore, leptin may be a valid neuroendocrinologic marker for the hypervigilant state of vulnerable people who have faced tremendous danger and uncertainty. Further studies are needed to examine the correlation between leptin levels and hypothalamic-pituitary-adrenal axis function, particularly the inverted diurnal serum leptin levels in hyperaroused subjects. PMID- 14747635 TI - Applying latent growth curve modeling to the investigation of individual differences in cardiovascular recovery from stress. AB - OBJECTIVE: This paper provides an introduction to latent growth curve (LGC) modeling, a modern method for analyzing data resulting from change processes such as cardiovascular recovery from stress. LGC models are superior to traditional approaches such as repeated measures analysis of variance and simple change scores. METHODS: The basic principles of LGC modeling are introduced and applied to data from 167 men and women whose systolic blood pressure was assessed before, during, and after the cold pressor and evaluated speech stressors and who had completed the Cook-Medley Hostility Inventory. RESULTS: The LGC models revealed that systolic blood pressure recovery follows a different nonlinear trajectory after speech relative to the cold pressor. The difference resulted not from the initial decline at the completion of the stressor, but from higher levels at the end of the stressor and slower rate of change in decline for the speech. Hostility predicted the trajectory for speech but not for cold pressor. This relationship did not differ as a function of gender, although men had larger systolic blood pressure responses than women to both stressors. CONCLUSIONS: LGC modeling yields an understanding of the processes and predictors of change that is not attainable through traditional statistical methods. Although our application concerns cardiovascular recovery from stress, LGC modeling has many other potential applications in psychosomatic research. PMID- 14747636 TI - Gender differences in health information needs and decisional preferences in patients recovering from an acute ischemic coronary event. AB - OBJECTIVE: This study examined gender differences in health information needs and decisional preferences after an acute ischemic coronary event (ICE). METHODS: Patients with ICE, recruited in 12 coronary intensive care units, completed a questionnaire on demographic, disease-related, and psychosocial topics. Six and 12 months later, they completed mailed follow-up questionnaires. RESULTS: Nine hundred six patients completed the baseline questionnaire, 541 (69%) completed the 6-month questionnaire, and 522 (64%) completed the 12-month questionnaire after hospital discharge. Men reported significantly more information received and greater satisfaction with healthcare practitioners meeting their information needs. Women wanted more information than men concerning angina and hypertension. Men wanted more information about sexual function and reported receiving more information about the role of each doctor, test results, treatments, cardiac rehabilitation, and how their families could support their lifestyle changes. Patients who reported receiving more information reported less depressive symptomatology and greater self-efficacy, healthcare satisfaction, and preventive health behaviors. Although most patients of both sexes preferred a shared decision-making role with their physician, the majority felt their doctor had made the main decisions. CONCLUSIONS: Patients after ICE, especially women, reported receiving much less information than they wanted from all health professionals. Most patients wanted a shared or autonomous treatment decision making role with their doctor, but only a minority experienced this. Clinicians must do better, because meeting patients' information needs and respecting their decisional preferences are shown to be associated with better self-efficacy, satisfaction, and health-promoting behavior. PMID- 14747637 TI - Low educational attainment, John Henryism, and cardiovascular reactivity to and recovery from personally relevant stress. AB - OBJECTIVE: The John Henryism hypothesis proposes that a high level of John Henryism (JH: high-effort coping with psychosocial demands) is predictive of hypertension at low but not high socioeconomic status (SES). The objectives of the present study were to determine whether high JH and low SES (education, income, job status, and job strain) were associated with increased cardiovascular responses to laboratory social stressors. METHODS: Subjects were 58 normotensive, healthy black men age 23 to 47 years. The procedure included the completion of psychosocial questionnaires and participation in a psychophysiological reactivity protocol. The reactivity protocol involved the following experimental tasks and associated recovery periods: an active speech task and an anger recall task. Measures of systolic blood pressure (SBP), diastolic blood pressure (DBP), heart rate (HR), and rate pressure product (RPP) were obtained continuously using a Finapres beat-to-beat blood pressure monitor throughout the reactivity protocol. RESULTS: At high JH, low (compared with high) education level was linked with higher DBP during anger recall and final recovery, higher SBP during final recovery, and higher HR and RPP during speech preparation and final recovery (p <.05). Among subjects with low education, high (vs. low) JH was associated with higher SBP, HR, and RPP during final recovery (p <.05). CONCLUSIONS: John Henryism may increase the risk of cardiovascular disease among people with low education by increased cardiovascular reactivity and prolonged recovery to stress. PMID- 14747638 TI - Acute stress affects heart rate variability during sleep. AB - OBJECTIVE: Although stress can elicit profound and lasting effects on sleep, the pathways whereby stress affects sleep are not well understood. In this study, we used autoregressive spectral analysis of the electrocardiogram (EKG) interbeat interval sequence to characterize stress-related changes in heart rate variability during sleep in 59 healthy men and women. METHODS: Participants (N = 59) were randomly assigned to a control or stress condition, in which a standard speech task paradigm was used to elicit acute stress in the immediate presleep period. EKG was collected throughout the night. The high frequency component (0.15-0.4 Hz Eq) was used to index parasympathetic modulation, and the ratio of low to high frequency power (0.04-0.15 Hz Eq/0.15-0.4 Hz Eq) of heart rate variability was used to index sympathovagal balance. RESULTS: Acute psychophysiological stress was associated with decreased levels of parasympathetic modulation during nonrapid eye movement (NREM) and rapid eye movement sleep and increased levels of sympathovagal balance during NREM sleep. Parasympathetic modulation increased across successive NREM cycles in the control group; these increases were blunted in the stress group and remained essentially unchanged across successive NREM periods. Higher levels of sympathovagal balance during NREM sleep were associated with poorer sleep maintenance and lower delta activity. CONCLUSIONS: Changes in heart rate variability associated with acute stress may represent one pathway to disturbed sleep. Stress-related changes in heart rate variability during sleep may also be important in association with chronic stressors, which are associated with significant morbidity and increased risk for mortality. PMID- 14747639 TI - Critical review of dietary caffeine and blood pressure: a relationship that should be taken more seriously. AB - OBJECTIVE: This review aims to determine whether, and to what extent, dietary caffeine may be a risk to cardiovascular health. METHODS: A critical review of the relevant experimental and epidemiologic literature was conducted, with particular reference to studies of caffeine and blood pressure (BP). RESULTS: There is extensive evidence that caffeine at dietary doses increases BP. However, concern that the drug may contribute to cardiovascular disease appears to have been dampened by (1) the belief that habitual use leads to the development of tolerance, and (2) confusion regarding relevant epidemiologic findings. When considered comprehensively, findings from experimental and epidemiologic studies converge to show that BP remains reactive to the pressor effects of caffeine in the diet. Overall, the impact of dietary caffeine on population BP levels is likely to be modest, probably in the region of 4/2 mm Hg. At these levels, however, population studies of BP indicate that caffeine use could account for premature deaths in the region of 14% for coronary heart disease and 20% for stroke. CONCLUSIONS: Current evidence supports the conclusion that the BP elevating effects of dietary caffeine may be contributing appreciably to population levels of cardiovascular mortality and morbidity. Accordingly, strategies for encouraging reduced dietary levels of caffeine deserve serious consideration. PMID- 14747640 TI - Predictors of employment of men with HIV/AIDS: a longitudinal study. AB - OBJECTIVE: To identify patterns and predictors of work status and number of hours employed in a group of men with human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome (AIDS). METHODS: A total of 141 participants had semiannual neuropsychiatric, psychosocial, and medical assessments over a period of 30 months. These six occasions provided the basis for identifying patterns of employment (part-time, full-time, or unemployed). Those who completed neuropsychological testing, introduced at visit 4, constitute the sample used to identify predictors of number of hours employed, using multiple regression analysis with mixed procedure. RESULTS: Over 30 months, 20% were continuously employed full-time, another 9% were continuously employed part-time, and 40% were continuously unemployed. Employment status changed for 31%: 4% who worked at baseline stopped, 13% started or increased their hours, 8% decreased their hours, and 6% showed a fluctuating pattern. The major parameters consistently associated with unemployment or partial employment, in order of influence, were financial (disability benefits), psychiatric (past/current diagnosis of major depression and/or dysthymia), medical (physical limitations), cognitive (executive function), and education. In contrast, age, ethnicity, laboratory markers of HIV illness status, vocational rank, and past or current substance dependence did not predict work status. CONCLUSIONS: Overall, those who worked continued to work. However, despite improved health, most men who were unemployed at study baseline did not return to work. Structure of disability benefits, lifetime depressive disorder, physical limitations, and impairment in some areas of cognitive function each appear to represent significant barriers to work. Returning to work is evidently difficult, and clinicians may keep this in mind when recommending leaving work unless medically necessary. Specific interventions and policy changes regarding disability benefits may be needed to promote return to work for people with HIV/AIDS whose health is restored and who contemplate re-employment. PMID- 14747641 TI - Impact of depressive mood on relapse in patients with inflammatory bowel disease: a prospective 18-month follow-up study. AB - OBJECTIVE: There is evidence of an interaction between psychological factors and activity of inflammatory bowel disease (IBD). We examined the influence of depressive mood and associated anxiety on the course of IBD over a period of 18 months in a cohort of patients after an episode of active disease. METHODS: In this prospective, longitudinal, observational study, 60 patients (37 women and 23 men) with clinically inactive IBD (Crohn disease, n = 47, 78%; ulcerative colitis, n = 13, 22%) were enrolled after a flare of disease. Psychological status, health-related quality of life (HRQOL), and disease activity were evaluated at baseline and then every 3 months for a period of 18 months by means of clinical and biological parameters, the Beck Depression Inventory (BDI), the Spielberger State-Trait Anxiety Inventory, the Inflammatory Bowel Disease Questionnaire, the Perceived Stress Questionnaire, and the Rating Form of Inflammatory Bowel Disease Patients Concerns. RESULTS: At baseline, depression (BDI > or = 13 points) was found in 17 of 60 (28%) patients. Thirty-two patients (59%) experienced at least one relapse during the 18 months of follow-up. Regression analysis showed a significant correlation between BDI scores at baseline and the total number of relapses after 12 (p <.01) and 18 months (p <.01) of follow-up. Furthermore, depression scores at baseline correlated with the time until the first recurrence of the disease (p <.05). Anxiety and low HRQOL were also related with more frequent relapses during follow-up (p <.05 and p <.01, respectively). CONCLUSIONS: Psychological factors such as a depressive mood associated with anxiety and impaired HRQOL may exert a negative influence on the course of IBD. Therefore, assessment and management of psychological distress should be included in clinical treatment of patients with IBD. PMID- 14747642 TI - Psychosocial factors and perceived severity of functional dyspeptic symptoms: a psychosocial interactionist model. AB - OBJECTIVE: A psychosocial interactionist model was presented to provide a systematic account of individual differences in perceived functional dyspeptic symptom severity. METHODS: In a population-based survey, 4038 Hong Kong subjects (age 18-80 years) were interviewed. Five hundred ninety interviewees (14.6%) met the diagnostic criteria for functional dyspepsia (FD), and 396 of them participated in this study. RESULTS: Results from multiple regression analyses revealed significant main effects of monitoring, emotional support, and coping flexibility on perceived FD symptom severity. A significant emotional support by coping flexibility interaction effect was also found. CONCLUSIONS: The present findings provided support for the psychosocial interactionist model in showing that (1) monitoring is a risk factor related to greater perceived symptom severity, (2) emotional support and coping flexibility are resource factors related to lower perceived symptom severity, and (3) the beneficial role of emotional support is present only among those higher in coping flexibility but not among those lower in coping flexibility. PMID- 14747643 TI - Cognitive processing among mothers of children undergoing bone marrow/stem cell transplantation. AB - OBJECTIVE: This study investigated the role of cognitive processing in maternal adjustment to a life-threatening pediatric medical procedure (bone marrow/stem cell transplantation: BMT/SCT). METHODS: Ninety-one mothers participated in structured interviews about their fears, intrusions, avoidance, and distress regarding their child's BMT/SCT at two time points: during their child's hospitalization and during his/her recovery. Structural equation modeling was used to determine the role of fears, intrusions, and avoidance in mothers' distress. RESULTS: Mothers' fears played a primary role in their adjustment to their child's transplantation. Intrusions mediated the relations of fears with distress at both time points. Mothers' avoidance of thoughts, feelings, and reminders of their child's illness during the child's transplantation was associated with their distress three months later. The child's risk for an unsuccessful transplantation outcome was not associated with mothers' fears or distress during the child's hospitalization, but was associated with mothers' distress during the child's posthospital course of recovery. CONCLUSIONS: The results of this study indicate the critical role of mothers' fears, intrusions, avoidance, and the child's transplant risk in maternal distress and have treatment implications for reducing maternal distress during pediatric transplantation. PMID- 14747644 TI - Stress response syndromes in women undergoing mammography: a comparison of women with and without a history of breast cancer. AB - OBJECTIVE: The purpose of this study was to assess the incidence, severity, and correlates of acute stress responses in women undergoing diagnostic mammographic surveillance and to explore the moderating impact of physician support on these symptoms. METHODS: Sixty-six female breast cancer outpatients (at least 12 months after diagnosis and primary treatment) and 69 healthy women undergoing mammographic surveillance completed measures of: acute stress response, somatization, trauma history, psychiatric history, social support, and physician satisfaction. RESULTS: Previous cancer, pre-mammography breast complaints, lower income, previous psychiatric medication use, greater instrumental support, greater somatization, greater perceived physician disengagement, and less perceived physician support were all associated with increased stress responses. Among women with a previous cancer diagnosis, those with greater distress reported higher levels of physician support. In contrast, among those without a previous cancer diagnosis, those with greater perceived physician support reported less distress. CONCLUSIONS: These findings suggest that cancer-related cues, such as follow-up surveillance, may trigger a sensitizing response in women with a previous cancer diagnosis. The association of distress with physician support may arise from the responsiveness of physicians to identified distress, from increased help-seeking behavior by those who are distressed, or both. The benefit of support provided by health care professionals to those at risk of developing stress response syndromes deserves further study. PMID- 14747645 TI - Mindfulness-based stress reduction and health-related quality of life: findings from a bilingual inner-city patient population. AB - OBJECTIVE: To determine whether completing a mindfulness-based stress reduction (MBSR) program would affect the general health, health-related quality of life, sleep quality, and family harmony of Spanish- and English-speaking medical patients at an inner-city health center. MATERIALS AND METHODS: An intervention group of 68 patients (48 Spanish-speaking and 20 English-speaking) completed the SF-36 Health Survey and two additional questions about sleep quality and family harmony before and after completing the 8-week MBSR program. A comparison group of 18 Spanish-speaking patients who received no intervention completed the same questionnaire at the same intervals. RESULTS: Sixty-six percent of the total intervention group completed the 8-week MBSR program. There was significant comorbidity of medical and mental health diagnoses among the intervention and comparison groups, with no differences in the mean number of diagnoses of the total intervention group, the comparison group, or the Spanish- or English speaking intervention subgroups. Compared with the comparison group, the intervention group showed statistically significant improvement on five of the eight SF-36 measures, and no improvement on the sleep quality or family harmony items. CONCLUSIONS: MBSR may be an effective behavioral medicine program for Spanish- and English-speaking inner-city medical patients. Suggestions are given for future research to help clarify the program's effectiveness for this population. PMID- 14747646 TI - Immunological effects of induced shame and guilt. AB - OBJECTIVE: To determine if inducing self-blame would lead to increases in shame and guilt as well as increases in proinflammatory cytokine activity and cortisol. Based on previous research and theory, it was hypothesized that induced shame would be specifically associated with elevations in proinflammatory cytokine activity. MATERIALS AND METHODS: Healthy participants were randomly assigned to write about traumatic experiences in which they blamed themselves (N = 31) or neutral experiences (N = 18) during three 20-minute experimental laboratory sessions over 1 week. Tumor necrosis factor-alpha receptor levels (sTNFalphaRII), an indicator of proinflammatory cytokine activity, beta2-microglobulin, cortisol (all obtained from oral fluids), and emotion were assessed prewriting and postwriting. RESULTS: Participants in the self-blame condition showed an increase in shame and guilt as well as an increase in sTNFalphaRII activity when compared with those in the control condition. Cortisol and beta2-microglobulin levels were unaffected by the procedures. Those individuals in the self-blame condition reporting the greatest increases in shame in response to the task showed the greatest elevations in proinflammatory cytokine activity, while levels of guilt and general negative emotion were unrelated to cytokine changes. CONCLUSION: These data suggest that inducing self-related emotions can cause changes in inflammatory products, and that shame may have specific immunological correlates. PMID- 14747647 TI - Alexithymia correlates with the size of the right anterior cingulate. AB - OBJECTIVE: The authors investigated a possible relationship between interindividual variability in anterior cingulate gyrus (ACG) morphology and alexithymia. MATERIALS AND METHODS: Magnetic resonance images were obtained in 100 healthy university graduates (51 female, 49 male; mean age 25.6 y). Surface area measurements of the ACG were performed on reformatted sagittal views in both hemispheres. The Toronto Alexithymia Scale (TAS-20) and the Temperament and Character Inventory (TCI) were administered. RESULTS: Right ACG surface area significantly correlated with TAS-20 total score in men (r = 0.37; p = 0.009) and in women (r = 0.30; p = 0.034). After controlling for three TCI subscales (harm avoidance, self-directedness, and self-transcendency), the correlation between TAS-20 total and right ACG became nonsignificant in women, but was only slightly reduced (r = 0.32; p = 0.032) in men. A linear regression model with right ACG as a dependent variable revealed brain volume, TCI-harm avoidance and TAS 20 total score as significant predictors in the total sample (explained proportion of total variation (EPTV) 37%). In men, beside brain volume, only TAS-20 total score showed a highly significant contribution (EPTV 41%), whereas in women only TCI harm avoidance was a significant predictor (EPTV 36%). CONCLUSIONS: The authors' findings indicate that there is a significant positive relation between the size of the right ACG and alexithymia as measured with the TAS in healthy subjects. This applies especially for men whereas in women ACG size is more associated with the subscale harm avoidance of the TCI. Our findings also suggest a partial lateralization of human emotion processing, especially negative emotion. PMID- 14747648 TI - The value of pseudoneurological symptoms for assessing psychopathology in primary care. AB - OBJECTIVE: This study sought to examine the relationship between pseudoneurological symptoms (PNS) and somatic and psychiatric symptom severity, physical functioning, and psychiatric comorbidity. METHODS: Interview and questionnaire data were obtained from 120 patients with somatization who participated in a study assessing the efficacy of cognitive-behavioral therapy. Measures elicited information on psychiatric diagnoses, anxiety and depressive symptom levels, somatic symptoms, and physical functioning. Statistical analyses examined the relationship between PNS and the diagnosis of somatization disorder, physical and psychiatric symptom severity, and psychiatric comorbidity. RESULTS: Roughly half of the sample had a history of four or more PNS. Results showed that having four or more PNS was not predictive of somatization disorder. However, having four or more PNS was found to be significantly correlated with the severity of anxiety, depression, somatic complaints, and physical dysfunction. These associations were identified while controlling for the symptom count of nonpseudoneurological symptoms, the presence of somatization disorder, and the presence of chronic painful physical conditions. In addition, having four or more PNS was significantly associated with a higher likelihood of receiving a diagnosis of major depression, dysthymia, panic disorder, and generalized anxiety disorder. CONCLUSIONS: A history of four or more PNS is common among somatizing patients in primary care and associated with a more severe clinical presentation, even after controlling for other factors known to be associated with severity. Four or more PNS may identify a distinct subgroup of somatization and serve as a clinical indicator for identifying psychiatric disorders in primary care. Future studies should explore the assessment of PNS using briefer measures. Furthermore, PNS should be evaluated with samples more representative of US primary care populations, as well as samples that include adequate representation from other ethnic backgrounds (eg, African-American, Asian, etc.). PMID- 14747649 TI - EEG asymmetry and mindfulness meditation. PMID- 14747650 TI - Alterations in brain and immune function produced by mindfulness meditation: three caveats. PMID- 14747651 TI - Brain masculinization requires androgen receptor function. AB - Testicular testosterone produced during a critical perinatal period is thought to masculinize and defeminize the male brain from the inherent feminization program and induce male-typical behaviors in the adult. These actions of testosterone appear to be exerted not through its androgenic activity, but rather through its conversion by brain aromatase into estrogen, with the consequent activation of estrogen receptor (ER)-mediated signaling. Thus, the role of androgen receptor (AR) in perinatal brain masculinization underlying the expression of male-typical behaviors remains unclear because of the conversion of testosterone into estrogen in the brain. Here, we report a null AR mutation in mice generated by the Cre loxP system. The AR-null mutation in males (AR(L-/Y)) resulted in the ablation of male-typical sexual and aggressive behaviors, whereas female AR-null homozygote (AR(L-/L-)) mice exhibited normal female sexual behaviors. Treatment with nonaromatizable androgen (5alpha-dihydrotestosterone, DHT) was ineffective in restoring the impaired male sexual behaviors, but it partially rescued impaired male aggressive behaviors in AR(L-/Y) mice. Impaired male-typical behaviors in ERalpha(-/-) mice were restored on DHT treatment. The role of AR function in brain masculinization at a limited perinatal stage was studied in AR(L-/L-) mice. Perinatal DHT treatment of females led to adult females sensitive to both 17beta estradiol and DHT in the induction of male-typical behaviors. However, this female brain masculinization was abolished by AR inactivation. Our results suggested that perinatal brain masculinization requires AR function and that expression of male-typical behaviors in adults is mediated by both AR-dependent and -independent androgen signaling. PMID- 14747653 TI - Evolution of document networks. AB - How does a network of documents grow without centralized control? This question is becoming crucial as we try to explain the emergent scale-free topology of the World Wide Web and use link analysis to identify important information resources. Existing models of growing information networks have focused on the structure of links but neglected the content of nodes. Here I show that the current models fail to reproduce a critical characteristic of information networks, namely the distribution of textual similarity among linked documents. I propose a more realistic model that generates links by using both popularity and content. This model yields remarkably accurate predictions of both degree and similarity distributions in networks of web pages and scientific literature. PMID- 14747652 TI - Long-term effects of culture of preimplantation mouse embryos on behavior. AB - Many procedures used in assisted reproductive technologies (ART) to treat human infertility entail culture of preimplantation embryos. Moreover, there is an increasing trend to culture embryos for longer periods of time before uterine transfer to identify the "best" embryos for transfer and to minimize multiple pregnancies. Embryo culture, however, can perturb embryo metabolism and gene expression, and the long-term consequences of culture are unknown. We have explored the behavioral consequences of embryo culture by using a 129S6/SvEvTac/C57BL/6J F(1) mouse model and find that adults derived from cultured embryos exhibit specific behavioral alterations in the elevated zero maze and Morris water maze tasks. PMID- 14747654 TI - Electrical wound-healing assay for cells in vitro. AB - Confluent cell monolayers in tissue culture are fragile and can easily be mechanically disrupted, often leaving an area devoid of cells. This opening in the cell sheet is then repopulated, because the cells on the fringe of the damage, which are no longer contact-inhibited, move into the available space. This mechanical disruption is often done deliberately in a "wound-healing" assay as a means to assess the migration of the cells. In such assays, a scrape is made in the cell layer followed by microscopy to monitor the advance of the cells into the wound. We have found that these types of assays can also be accomplished electrically. In this approach, cells growing on small electrodes and monitored by using electric cell-substrate impedance sensing are subjected to currents, resulting in severe electroporation and subsequent cell death. After this invasive treatment, the electrode's impedance is again monitored to chart the migration and ultimate healing of the wound. We report here that this procedure to study cell behavior is both highly reproducible, quantitative, and provides data similar to that acquired with traditional measurements. PMID- 14747655 TI - Cytogenetic evidence for asexual evolution of bdelloid rotifers. AB - DNA sequencing has shown individual bdelloid rotifer genomes to contain two or more diverged copies of every gene examined and has revealed no closely similar copies. These and other findings are consistent with long-term asexual evolution of bdelloids. It is not entirely ruled out, however, that bdelloid genomes consist of previously undetected pairs of sequences so similar as to be identical over the regions sequenced, as might result if bdelloids were highly inbred sexual diploids or polyploids. Here, we employ fluorescent in situ hybridization with cosmid probes to determine the copy number and chromosomal distribution of the heat shock gene hsp82 and adjacent sequences in the bdelloid Philodina roseola. We conclude that the four copies identified by sequencing are the only ones present and that each is on a separate chromosome. Bdelloids therefore are not highly homozygous sexually reproducing diploids or polyploids. PMID- 14747656 TI - Stabilities of folding of clustered, two-repeat fragments of spectrin reveal a potential hinge in the human erythroid spectrin tetramer. AB - The large size of spectrin, the flexible protein promoting reversible deformation of red cells, has been an obstacle to elucidating the molecular mechanism of its function. By studying cloned fragments of the repeating unit domain, we have found a correspondence between positions of selected spectrin repeats in a tetramer with their stabilities of folding. Six fragments consisting of two spectrin repeats were selected for study primarily on the basis of the predicted secondary structures of their linker regions. Fragments with a putatively helical linker were more stable to urea- and heat-induced unfolding than those with a putatively nonhelical linker. Two of the less stably folded fragments, human erythroid alpha-spectrin repeats 13 and 14 (HEalpha13,14) and human erythroid beta-spectrin repeats 8 and 9 (HEbeta8,9), are located opposite each other on antiparallel spectrin dimers. At least partial unfolding of these repeats under physiological conditions indicates that they may serve as a hinge. Also less stably folded, the fragment of human erythroid alpha-spectrin repeats 4 and 5 (HEalpha4,5) lies opposite the site of interaction between the partial repeats at the C- and N-terminal ends of beta- and alpha-spectrin, respectively, on the opposing dimer. More stably folded fragments, human erythroid alpha-spectrin repeats 1 and 2 (HEalpha1,2) and human erythroid alpha-spectrin repeats 2 and 3 (HEalpha2,3), lie nearly opposite each other on antiparallel spectrin dimers of a tetramer. These clusterings along the spectrin tetramer of repeats with similar stabilities of folding may have relevance for spectrin function, particularly for its well known flexibility. PMID- 14747657 TI - Heme regulates the dynamic exchange of Bach1 and NF-E2-related factors in the Maf transcription factor network. AB - Small Maf proteins serve as dual-function transcription factors through an exchange of their heterodimerization partners. For example, as heterodimers with hematopoietic cell-specific p45 NF-E2 or NF-E2-related factors (Nrf), they activate the beta-globin or antioxidative stress enzyme heme oxygenase 1 (HO-1) genes, respectively. In contrast, together with Bach1, they repress these same genes. However, the signals leading to this partner exchange are not known. Using chromatin immunoprecipitation assays in NIH 3T3 cells, we show that heme, an inducer of ho-1, promotes displacement of Bach1 from the MafK-occupied ho-1 enhancers, which is followed by Nrf2 binding to these elements. Whereas histone H3 at the ho-1 enhancers and promoter is hyperacetylated irrespective of gene activity, exposure of cells to heme results in de novo hyperacetylation and hypermethylation of histone H3 in the transcribed region. These data indicate that, under normal conditions, the chromatin structure of ho-1 is in a preactivation state, but transcription is repressed by Bach1. Heme induces switching of Maf dimers, resulting in ho-1 expression. Heme also promotes displacement of Bach1 from the beta-globin locus control region without affecting MafK binding in murine erythroleukemia cells. Thus, heme functions as a signaling molecule for gene expression in higher eukaryotes. PMID- 14747658 TI - Centrosome maturation: measurement of microtubule nucleation throughout the cell cycle by using GFP-tagged EB1. AB - Understanding how cells regulate microtubule nucleation during the cell cycle has been limited by the inability to directly observe nucleation from the centrosome. To view nucleation in living cells, we imaged GFP-tagged EB1, a microtubule tip binding protein, and determined rates of nucleation by counting the number of EB1 GFP comets emerging from the centrosome over time. Nucleation rate increased 4 fold between G(2) and prophase and continued to rise through anaphase and telophase, reaching a maximum of 7 times interphase rates. We tested several models for centrosome maturation, including gamma-tubulin recruitment and increased centrosome size. The centrosomal concentration of gamma-tubulin reached a maximum at metaphase, and centrosome size increased through anaphase, whereas nucleation remained high through telophase, implying the presence of additional regulatory processes. Injection of anti-gamma-tubulin antibodies significantly blocked nucleation during metaphase but was less effective during anaphase, suggesting that a nucleation mechanism independent of gamma-tubulin contributes to centrosome function after metaphase. PMID- 14747659 TI - Early onset of neoplasia in the prostate and skin of mice with tissue-specific deletion of Pten. AB - PTEN is a tumor suppressor gene mutated in various advanced human neoplasias, including glioblastomas and prostate, breast, endometrial, and kidney cancers. This tumor suppressor is a lipid phosphatase that negatively regulates cell survival and proliferation mediated by phosphatidylinositol 3-kinase/protein kinase B signaling. Using the Cre-loxP system, we selectively inactivated Pten in murine tissues in which the MMTV-LTR promoter is active, resulting in hyperproliferation and neoplastic changes in Pten-null skin and prostate. These phenotypes had early onset and were completely penetrant. Abnormalities in Pten mutant skin consisted of mild epidermal hyperplasia, whereas prostates from these mice exhibited high-grade prostatic intraepithelial neoplasia (HGPIN) that frequently progressed to focally invasive cancer. These data demonstrate that Pten is an important physiological regulator of growth in the skin and prostate. Further, the early onset of HGPIN in Pten mutant males is unique to this animal model and implicates PTEN mutations in the initiation of prostate cancer. Consistent with high PTEN mutation rates in human prostate tumors, these data indicate that PTEN is a critical tumor suppressor in this organ. PMID- 14747660 TI - Divergent gene copies in the asexual class Bdelloidea (Rotifera) separated before the bdelloid radiation or within bdelloid families. AB - Rotifers of the asexual class Bdelloidea are unusual in possessing two or more divergent copies of every gene that has been examined. Phylogenetic analysis of the heat-shock gene hsp82 and the TATA-box-binding protein gene tbp in multiple bdelloid species suggested that for each gene, each copy belonged to one of two lineages that began to diverge before the bdelloid radiation. Such gene trees are consistent with the two lineages having descended from former alleles that began to diverge after meiotic segregation ceased or from subgenomes of an alloploid ancestor of the bdelloids. However, the original analyses of bdelloid gene-copy divergence used only a single outgroup species and were based on parsimony and neighbor joining. We have now used maximum likelihood and Bayesian inference methods and, for hsp82, multiple outgroups in an attempt to produce more robust gene trees. Here we report that the available data do not unambiguously discriminate between gene trees that root the origin of hsp82 and tbp copy divergence before the bdelloid radiation and those which indicate that the gene copies began to diverge within bdelloid families. The remarkable presence of multiple diverged gene copies in individual genomes is nevertheless consistent with the loss of sex in an ancient ancestor of bdelloids. PMID- 14747661 TI - An integrated reverse functional genomic and metabolic approach to understanding orotic acid-induced fatty liver. AB - In functional genomics, DNA microarrays for gene expression profiling are increasingly being used to provide insights into biological function or pathology. To better understand the significance of the multiple transcriptional changes across a time period, the temporal changes in phenotype must be described. Orotic acid-induced fatty liver disease was investigated at the transcriptional and metabolic levels using microarrays and metabolic profiling in two strains of rats. High-resolution 1H-NMR spectroscopic analysis of liver tissue indicated that Kyoto rats compared with Wistar rats are predisposed to the insult. Metabolite analysis and gene expression profiling following orotic acid treatment identified perturbed metabolic pathways, including those involved in fatty acid, triglyceride, and phospholipid synthesis, beta-oxidation, altered nucleotide, methyl donor, and carbohydrate metabolism, and stress responses. Multivariate analysis and statistical bootstrapping were used to investigate co responses with transcripts involved in metabolism and stress responses. This reverse functional genomic strategy highlighted the relationship between changes in the transcription of stearoyl-CoA desaturase 1 and those of other lipid related transcripts with changes in NMR-derived lipid profiles. The results suggest that the integration of 1H-NMR and gene expression data sets represents a robust method for identifying a focused line of research in a complex system. PMID- 14747662 TI - DNA microarray analysis of gene expression in human optic nerve head astrocytes in response to hydrostatic pressure. AB - There is clinical and experimental evidence that elevated intraocular pressure (IOP), a mechanical stress, is involved in the pathogenesis of glaucomatous optic neuropathy. The mechanism by which astrocytes in the optic nerve head (ONH) respond to changes in IOP is under study. Gene transcription by ONH astrocytes exposed either to 60 mmHg hydrostatic pressure (HP) or control ambient pressure (CP) for 6, 24, and 48 h was compared using Affymetrix GeneChip microarrays to identify HP-responsive genes. Data were normalized across arrays within each gene. A linear regression model applied to test effect of time and HP on changes in expression level identified 596 genes affected by HP over time. Using GeneSpring analysis we selected genes whose average expression level increased or decreased more than 1.5-fold at 6, 24, or 48 h. Expression of selected genes was confirmed by real-time RT-PCR; protein levels were detected by Western blot. Among the genes highly responsive to HP were those involved in signal transduction, such as Rho nucleotide exchange factors, Ras p21 protein activator, tyrosine kinases and serine threonine kinases, and genes involved in transcriptional regulation, such as c-Fos, Egr2, and Smad3. Other genes that increased expression included ATP-binding cassettes, solute carriers, and genes associated with lipid metabolism. Among the genes that decreased expression under HP were genes encoding for dual activity phosphatases, transcription factors, and enzymes involved in protein degradation. These HP-responsive genes may be important in the establishment and maintenance of the ONH astrocyte phenotype under conditions of elevated IOP in glaucoma. PMID- 14747663 TI - Energy homeostasis, obesity and eating disorders: recent advances in endocrinology. AB - Health problems resulting from obesity could offset many of the recent health gains achieved by modern medicine, and obesity may replace tobacco as the number one health risk for developed societies. An estimated 300,000 deaths per year and significant morbidity are directly attributable to obesity, mainly due to heart disease, diabetes, cancer, asthma, sleep apnea, arthritis, reproductive complications and psychological disturbances. In parallel with the increasing prevalence of obesity, there has been a dramatic increase in the number of scientific and clinical studies on the control of energy homeostasis and the pathogenesis of obesity to further our understanding of energy balance. It is now recognized that there are many central and peripheral factors involved in energy homeostasis, and it is expected that the understanding of these mechanisms should lead to effective treatments for the control of obesity. This brief review discusses the potential role of several recently discovered molecular pathways involved in the control of energy homeostasis, obesity and eating disorders. PMID- 14747664 TI - Isomers of conjugated linoleic acid differ in their effects on angiogenesis and survival of mouse mammary adipose vasculature. AB - Dietary conjugated linoleic acid (CLA) is a cancer chemopreventive agent that has been shown to inhibit angiogenesis in vivo and in vitro, and to decrease vascular endothelial growth factor (VEGF) and Flk-1 concentrations in the mouse mammary gland. To determine which isomer mediates the antiangiogenic effects of CLA in vivo, the effects of diets supplemented with 5 or 10 g/kg c9,t11- or t10,c12-CLA isomers were compared in CD2F1Cr mice. Both isomers inhibited functional vascularization of a matrigel pellet in vivo and decreased serum VEGF concentrations; the t10,c12 isomer also decreased the proangiogenic hormone leptin (P < 0.05). Additionally, the t10,c12 isomer, but not c9,t11-CLA, rapidly induced apoptosis of the white and brown adipocytes as well as the preexisting supporting vasculature of the mammary fat pad (P < 0.05). Independent of this isomer-specific adipose apoptotic effect, both isomers induced a rapid and reversible decrease in the diameter of the unilocular adipocytes (P < 0.05). The ability of both CLA isomers to inhibit angiogenesis in vivo may contribute to their ability to inhibit carcinogenesis. Moreover, we propose that each CLA isomer uniquely modifies the mammary stromal "soil" in a manner that is useful for chemoprevention of breast cancer. PMID- 14747665 TI - Point mutations alter the cellular distribution of the human folate receptor in cultured Chinese hamster ovary cells. AB - Diminished cellular need for folate results in decreased function of the human folate receptor (FR) but increased expression of this protein in cells grown at different rates. Much of this FR is intracellular and not available for vitamin transport, raising the following question: what is the function of this excess receptor? In this study, we characterized the effects of three point mutations on FR regulation in Chinese hamster ovary cells stably transfected to express either wild-type receptor or FR containing mutations at positions 67(S-->P),144(E-->D), and/or 201(N-->D). The 201(N-->D) FR responded functionally like the wild-type receptor but was localized predominantly at the cell surface (>90% vs. <40% for wild-type). This mutation disrupted a N-linked glycosylation site and generated a partially deglycosylated receptor. The 67(S-->P) mutation also shifted the cellular distribution such that more FR was surface accessible ( approximately 80%) but did not affect glycosylation. Because previous results showed that these mutations influence the conformation of FR, our findings suggest that structural changes in the receptor facilitate its trafficking to the cell surface. FR containing the 67(S-->P) mutation with either a 144(E-->D) or 201(N-->D) change was not processed from the high-mannose to complex glycoform but was still transported to the cell surface and able to transport folates. Thus, conformational changes introduced by specific point mutations can influence FR processing and/or trafficking to the cell surface. Furthermore, the fact that mutated FR can be trafficked to the cell surface more efficiently suggests that the native receptor may be retained intracellularly to perform some function there. PMID- 14747666 TI - 3-Hydroxypropionic acid and methylcitric acid are not reliable indicators of marginal biotin deficiency in humans. AB - In two studies comprising 10 and 11 subjects, respectively, marginal biotin deficiency was induced experimentally by an egg-white diet in healthy men and women. The following urinary organic acids were assessed for their usefulness in detecting marginal biotin status: 1) 3-hydroxypropionic acid and methylcitric acid, organic acids that reflect decreased activity of the biotin-dependent enzyme propionyl-CoA carboxylase and 2) methylcrotonylglycine and isovalerylglycine, organic acids that reflect decreased activity of methylcrotonyl-CoA carboxylase. Mean 3-hydroxypropionic acid excretion rates remained normal during biotin depletion in both studies. By the end of the depletion period, 3-hydroxypropionic acid excretion identified only 5 of 21 marginally deficient subjects. Mean methylcitric acid excretion increased (P < 0.0001) in the first study but not in the second. Mean methylcrotonylglycine excretion increased in each study (P < 0.004 and P < 0.05, respectively); methylcrotonylglycine excretion identified 13 of 21 marginally deficient subjects. Mean isovalerylglycine excretion increased only in the first study (P = 0.006) and identified only 6 of 21 deficient subjects. We conclude that none of these organic acids is as sensitive an indicator of marginal biotin deficiency as 3-hydroxyisovaleric acid, which reflects decreased methylcrotonyl-CoA carboxylase. PMID- 14747667 TI - A low protein diet alters gene expression in rat pancreatic islets. AB - Insulin secretion is regulated mainly by circulating nutrients, particularly glucose, and is also modulated by hormonal and neuronal inputs. Nutritional alterations during fetal and early postnatal periods, induced by either low protein or energy-restricted diets, produce beta-cell dysfunction. As a consequence, insulin secretion in response to different secretagogues is reduced, as is the number of beta-cells and the size and vascularization of islets. In this study, we used a cDNA macroarray technique and RT-PCR to assess the pattern of gene expression in pancreatic islets from rats fed isocaloric low (6 g/100 g, LP) and normal (17 g/100 g, NP) protein diets, after weaning. Thirty-two genes related to metabolism, neurotransmitter receptors, protein trafficking and targeting, intracellular kinase network members and hormones had altered expression (up- or down-regulated). RT-PCR confirmed the macroarray results for five selected genes, i.e., clusterin, secretogranin II precursor, eukaryotic translation initiation factor 2, phospholipase A(2) and glucose transporter. Thus, cDNA macroarray analysis revealed significant changes in the gene expression pattern in rats fed a low protein diet after weaning. The range of proteins affected indicated that numerous mechanisms are involved in the intracellular alterations in the endocrine pancreas, including impaired glucose induced insulin secretion. PMID- 14747668 TI - Isoenergetic dietary protein restriction decreases myosin heavy chain IIx fraction and myosin heavy chain production in humans. AB - The synthesis of muscle protein is restrained during dietary protein restriction. This is widely understood to vary quantitatively with the degree of nutritional deprivation, but there has been little discussion of qualitative changes in muscle protein deriving from dietary protein restriction. We studied 14 healthy subjects in a 2-sample study. Subjects were randomly assigned to a diet providing an ample, American-style protein intake (1.67 g. kg fat-free mass(-1). d(-1)) or a diet approximating the mean minimum adult protein requirement (0.71 g. kg fat free mass(-1). d(-1)). We found that consumption of an isoenergetic diet at the mean adult minimum protein requirement for 4 wk produced an 81% lower fractional synthesis rate of myosin heavy chain (MHC) proteins in vastus lateralis muscle than did consumption of an ample protein diet (P = 0.05). Protein deprivation altered the skeletal muscle myosin composition such that the proportion of the total myosin content represented by fast-twitch MHC IIx was 51% lower than with ample intake (P = 0.013). The steady state content of MHC IIx messenger RNA (mRNA) did not differ in subjects consuming the minimum requirement of protein, suggesting that the reduced proportion of MHC IIx arises from posttranscriptional events. A 68% lower rate of 3-methylhistidine excretion with protein restriction (P < 0.01) suggests that myofibrillar protein degradation was lower. We conclude that dietary amino acid scarcity produces a change in myosin isoform distribution via posttranscriptional mechanisms. The relative contribution of inhibited myosin synthesis and inhibited degradation to the altered myosin isoform composition remains unknown. This has implications for the mechanisms by which amino acids govern muscle protein composition in vivo, and further exploration is required. PMID- 14747669 TI - Dietary energy density is associated with increased intake in free-living humans. AB - Dietary energy density markedly influences the daily dietary intake of humans. The present study examined the relation of energy density to dietary intake and body size in 371 male and 581 female adults. The subjects were free-living and provided a detailed record of their everyday food and drink intake in 7-d food diaries. The reported diets were analyzed for the relation of energy density to body size and to per-meal, daily, and weekly dietary intake. High energy density was associated with greater (P < 0.001) total intake, especially of fat, when intake was correlated with energy density (r = 0.26, P < 0.001) and when the daily intake of individual participants was correlated with their daily dietary density (r = 0.66, P < 0.001). These results occurred regardless of sex, low reporting, or the inclusion of drinks in the energy density calculations. High energy density was also associated with a high rate of intake (r = 0.46, P < 0.001) and large meal sizes (r = 0.61, P < 0.001). However, energy density was not correlated with body size, height, weight, or body mass index. High energy density appears to be related to greater overall intake in the short-term, but there may be compensation over the long term, with no net effect on body size. PMID- 14747670 TI - Plasma triacylglycerol and HDL cholesterol concentrations confirm self-reported changes in carbohydrate and fat intakes in women in a diet intervention trial. AB - Diet intervention trials are currently testing whether reduced fat intake can reduce the risk and progression of breast cancer. Energy from dietary fat is generally replaced by energy from carbohydrate in these studies, and altering the proportion of energy from dietary carbohydrate and fat has been shown to affect plasma lipid concentrations in controlled feeding studies. The purpose of this study was to examine the effect of increased carbohydrate and reduced fat intakes on plasma lipids in a randomized, controlled trial that is testing the effect of diet modification on risk for recurrence and survival in women previously treated for breast cancer. Plasma concentrations of lipids and related factors were measured at enrollment and 1-y follow-up in 393 women enrolled in the trial. Dietary goals for the intervention group focused on an increase in vegetable, fruit and fiber intakes, and reduced fat intake. Women assigned to the intervention group significantly reduced fat intake (from 28.1 to 21.0% of energy), and significantly increased intakes of carbohydrate (from 56.9 to 65.3% of energy) and fiber (from 21.0 to 29.6 g/d) (P < 0.05). Body weight did not change significantly in either study group. A small but significant increase in fasting plasma triacylglycerol concentration, and decreases in HDL cholesterol and apoprotein-A1 concentrations, were observed in the intervention group (P < 0.05) but not in the comparison group. Changes in total cholesterol, LDL cholesterol, apoprotein-B, lipoprotein (a), and insulin concentrations, and in the LDL cholesterol/HDL cholesterol ratio, were not observed in either group. The lipid responses that were observed in this study provide biological evidence that validates the self-reported change in dietary intakes of fat and carbohydrate in response to the intervention efforts. The degree of change in these lipid concentrations was small and does not suggest increased cardiovascular disease risk. PMID- 14747671 TI - Low dose daily iron supplementation improves iron status and appetite but not anemia, whereas quarterly anthelminthic treatment improves growth, appetite and anemia in Zanzibari preschool children. AB - Iron deficiency and helminth infections are two common conditions of children in developing countries. The consequences of helminth infection in young children are not well described, and the efficacy of low dose iron supplementation is not well documented in malaria-endemic settings. A 12-mo randomized, placebo controlled, double-blind trial of 10 mg daily iron and/or mebendazole (500 mg) every 3 mo was conducted in a community-based sample of 459 Zanzibari children age 6-71 mo with hemoglobin > 70 g/L at baseline. The trial was designed to examine treatment effects on growth, anemia and appetite in two age subgroups. Iron did not affect growth retardation, hemoglobin concentration or mild or moderate anemia (hemoglobin < 110 g/L or < 90 g/L, respectively), but iron significantly improved serum ferritin and erythrocyte protoporphyrin. Mebendazole significantly reduced wasting malnutrition. but only in children <30 mo old. The adjusted odds ratios (AORs) for mebendazole in this age group were 0.38 (95% CI: 0.16, 0.90) for weight-for-height less than -1 Z-score and 0.29 (0.09, 0.91) for small arm circumference. In children <24 mo old, mebendazole also reduced moderate anemia (AOR: 0.41, 0.18, 0.94). Both iron and mebendazole improved children's appetite, according to mothers' report. In this study, iron's effect on anemia was limited, likely constrained by infection, inflammation and perhaps other nutrient deficiencies. Mebendazole treatment caused unexpected and significant reductions in wasting malnutrition and anemia in very young children with light infections. We hypothesize that incident helminth infections may stimulate inflammatory immune responses in young children, with deleterious effects on protein metabolism and erythropoiesis. PMID- 14747672 TI - Nutrition counseling training changes physician behavior and improves caregiver knowledge acquisition. AB - Physician behavior and caregiver retention of nutrition advice were examined as potential mediating factors in the success of a nutrition counseling efficacy trial in Pelotas, Brazil, which reduced growth faltering in children 12-24 mo old. After pair-matching on socioeconomic status and nutrition indicators, municipal health centers were randomly assigned to an intervention group, in which physicians were trained with an IMCI-derived (Integrated Management of Childhood Illness) nutrition counseling protocol, or to a control group, without continuing education in nutrition. In a substudy of the larger trial, direct observation of consultations, followed by home interviews with mothers, provided data on physician counseling behavior and mothers' retention of nutrition advice. Trained providers were more likely to engage in nutrition counseling (P < 0.013) and to deliver more extensive advice (P < 0.02). They also used communication skills designed to improve rapport and ensure that mothers understood the advice (P < 0.01). Mothers who received advice from trained providers had high rates of recalling the messages on specific foods (95 vs.27%; P < 0.01) and feeding practice and food preparation recommendations (90 vs. 20%; P < 0.01), whereas the proportions of the messages recalled on breast-feeding (60% vs. 30%) did not differ significantly (P < 0.20). The training course contained several elements that may explain why intervention group mothers were better able to recall nutrition advice. These include locally appropriate messages, tools for assessing individual problems, and counseling skills. PMID- 14747673 TI - Blood lead, anemia, and short stature are independently associated with cognitive performance in Mexican school children. AB - Lead exposure and nutritional factors are both associated with cognitive performance. Lead toxicity and nutritional status are also associated with each other. We examined whether nutritional status variables account for part or all of the association between cognitive performance and lead exposure. First-grade children (n = 724) ages 6-8 y, attending Mexican public schools located in the vicinity of a metal foundry were asked to participate and 602 enrolled in the study. Blood lead, iron status, anemia, anthropometry, and cognitive function were assessed. Results from 7 standardized tests are presented here. The mean blood lead concentration was 11.5 +/- 6.1 micro g/dL (0.56 +/- 0.30 micro mol/L) and 50% of the children had concentrations >10 micro g/dL (0.48 micro mol/L). The prevalence of mild anemia (<124 g/L) was low (10%) and stunting (<2 SD) was nonexistent (2.3%). In bivariate analyses, lead was negatively associated with 4 cognitive tests and was also inversely correlated with iron status, height-for age Z scores, and head circumference. In multivariate models, the association between lead and cognitive performance was not strongly affected by nutritional variables, suggesting that the relation of lead to cognition is not explained by lead's relation to iron deficiency anemia or growth retardation. In multivariate models, hemoglobin concentration was also positively associated with Peabody Picture Vocabulary Test and Number Sequencing performance, whereas serum ferritin was negatively related to the Coding subscale of the Wechsler Intelligence Scales for Children-Revised Mexican Version (WISC-RM). PMID- 14747674 TI - Long-term food stamp program participation is differentially related to overweight in young girls and boys. AB - This paper examines the relation between long-term Food Stamp Program (FSP) participation and overweight in children using data on children from the National Longitudinal Survey of Youth 1979 Child Sample. A child was categorized as overweight if his or her BMI was >/= the 95th percentile of sex- and age-specific BMI. The data were arranged as a panel with multiple observations per child, and the preferred models of overweight included long-term FSP participation, additional demographic, socioeconomic, and environmental characteristics, and child fixed effects. Child fixed effects were used to take into account unobserved differences across children that did not vary over time. The models were estimated separately for younger (5-11 y old) and older (12-18 y old) children. In Ordinary Least Squares models, long-term FSP participation was positively and significantly related to overweight in young girls (P = 0.048), and negatively and significantly related to overweight in young boys (P = 0.100). Compared with girls and boys whose families did not participate in the FSP during the previous 5 y, FSP participation during all of the previous 5 y was associated with a 42.8% increase for young girls and a 28.8% decrease for young boys in the predicted probability of overweight. Long-term FSP participation was not significantly related to overweight in older children. Although these models did not control for food insecurity, the potential role of food insecurity in FSP participation was considered in the interpretation of the relation between FSP participation and child weight. PMID- 14747675 TI - The poor bioavailability of elemental iron in corn masa flour is not affected by disodium EDTA. AB - The most sustainable way to eradicate iron deficiency is through food fortification. Elemental iron powders are commonly utilized as fortificants due to their low cost and few sensory problems. However, their bioavailability is unknown. Our goals were to measure the bioavailability of elemental iron in Mexican style corn masa flour tortillas and to evaluate the effects of Na(2)EDTA. We used a stable isotope of H(2)-reduced iron powder, with and without Na(2)EDTA in tortillas prepared with corn masa flour. Two groups of 5- to 7-y-old children (n = 12/group) were fed tortillas to which was added 3 mg/100 g of H(2)-reduced (58)Fe with a mean particle size of 15 micro m. In one group, Na(2)EDTA was incorporated at a ratio of 1:2 mol/mol. The next day, (57)Fe ascorbate was given as a reference dose. After 14 d, blood samples were analyzed for isotopic enrichment. When normalized to 40% absorption of the reference dose, the geometric mean (+/-range 1 SD) bioavailability of reduced iron in tortilla was 3.8% (2.7-5.3). The addition of Na(2)EDTA, tended to increase it (P = 0.18) to 5.1% (2.8-9.2). This observed low absorption was compounded by the use of iron isotopes with smaller particle size (mean diameter 15 micro m) than typical of commercial elemental iron powder (<45 micro m). We conclude that H(2)-reduced iron powder is an ineffective fortificant in corn tortillas. PMID- 14747676 TI - Use of combined measures from capillary blood to assess iron deficiency in rural Kenyan children. AB - Community-based surveys of iron deficiency (ID) require simple, accurate methods that can be used in remote areas. The objective of this study was to assess iron status in rural Kenya using "field-friendly" methods for capillary blood, including an improved dried blood spot assay for transferrin receptor (TfR). A single finger stick was used to obtain capillary blood from 275 school-age children. Whole blood was applied directly to filter paper, dried, and later analyzed for TfR, as well as C-reactive protein (CRP), an acute-phase protein that serves as a general marker of inflammation. Capillary blood was also used to measure hemoglobin (Hb) concentration and the ratio of zinc protoporphyrin to heme (ZPP:H). The Hb concentration alone provides the lowest estimate of the prevalence of ID (8.0%). Because ZPP:H is reported to be elevated in the presence of inflammation, we constructed a preliminary diagnostic model based on elevated ZPP:H and normal CRP level, estimating the prevalence of ID at 25.9%. When TfR is added to a multiple criteria model (elevated ZPP:H in the absence of elevated CRP and/or elevated TfR level) the prevalence of ID is estimated to be 31.2%. This study demonstrates the diagnostic utility of combining TfR with other indexes of iron status, enabling the detection of ID in both the presence and absence of infection. Furthermore, this study is the first field application of TfR blood spot methods, and it demonstrates their feasibility in remote field settings. PMID- 14747677 TI - Repletion with (n-3) fatty acids reverses bone structural deficits in (n-3) deficient rats. AB - (n-3) PUFA deficiency and repletion effects on bone mechanical properties have not been examined. The primary research aim was to evaluate whether changes in the fatty acid composition of bone tissue compartments previously reported to influence bone formation rates would affect bone modeling and mechanical properties. In this investigation, three groups of rats were studied, second generation (n-3)-deficient, (n-3)-repleted, and a control (n-3)-adequate. The (n 3)-adequate diet contained alpha-linolenic acid [LNA, 18:3(n-3), 2.6% of total fatty acids] and docosahexaenoic acid [DHA, 22:6(n-3), 1.3% of total fatty acids]. Fatty acid composition of the hindlimb tissues (bone and muscle) of chronically (n-3)-deficient rats revealed a marked increase in (n-6) PUFA [20:4(n 6), 22:4(n-6), and 22:5(n-6)] and a corresponding decrease in (n-3) PUFA [18:3(n 3), 20:5(n-3), 22:5(n-3) and 22:6(n-3)]. Measurement of bone mechanical properties (energy to peak load) of tibiae showed that (n-3) deficiency diminished structural integrity. Rats repleted with (n-3) fatty acids demonstrated accelerated bone modeling (cross-sectional geometry) and an improved second moment in tibiae compared with control (n-3)-adequate rats after 28 d of dietary treatment. This study showed that repletion with dietary (n-3) fatty acids restored the ratio of (n-6)/(n-3) PUFA in bone compartments and reversed compromised bone modeling in (n-3)-deficient rats. PMID- 14747678 TI - The mechanisms for regulating absorption of Fe bis-glycine chelate and Fe ascorbate in caco-2 cells are similar. AB - Inorganic iron (Fe) absorption from the diet is controlled mainly in the intestinal tract where apical Fe uptake is inversely related to the Fe content in the enterocyte. Iron bis-glycine chelate is an iron compound that may be absorbed by a mechanism different from the regulated nonheme Fe pathway. Because Fe bis glycine chelate is used increasingly as an Fe fortificant in foods, the critical question is whether this compound is a safe Fe supplement. We compared apical Fe uptake and transepithelial transport offered either as (59)Fe bis-glycine chelate or a (59)Fe-ascorbate (Fe-AA) complex in Caco-2 cells, as a model of human intestinal epithelia, grown in different Fe concentrations in the media (0.5, 5 and 20 micro mol/L Fe). Apical Fe uptake from (59)Fe-AA and (59)Fe bis-glycine chelate did not differ nor did transepithelial transport rates. The rate of (59)Fe uptake decreased with increasing intracellular Fe concentration (P < 0.001), an indication of a common absorption regulatory mechanism. We also evaluated the effect of an excess of Fe (100 micro mol/L) provided as Fe bis glycine chelate or Fe-AA on the incorporation of 1 micro mol/L (55)Fe-AA into Fe replete Caco-2 cells. The inhibition of Fe bis-glycine chelate on the absorption of the extrinsic tag of (55)Fe-AA (87.5%) did not differ from that of Fe added as Fe-AA (86.8%). These results suggest that Fe derived from Fe bis-glycine chelate and Fe-AA have similar regulatory absorption mechanisms. PMID- 14747679 TI - Nondigestible oligosaccharides increase calcium absorption and suppress bone resorption in ovariectomized rats. AB - Nondigestible oligosaccharides (NDO) including inulin and fructooligosaccharides (FOS) have been reported to stimulate calcium absorption. Here we report the effect of a mixture of inulin and FOS (Raftilose Synergy 1, Orafti) on calcium and bone metabolism in ovariectomized (OVX) rats. OVX rats (6 mo old) were fed a semipurified diet for 3 mo in our animal care laboratory for stabilization after ovariectomy. They were then divided into two groups (n = 13/group) and fed either a control or a NDO-supplemented diet (55 g/kg) for 21 d. Catheters were placed in their jugular veins. After 2 d, a tracer ((45)Ca) was administered by gavage or i.v. and blood was sampled for up to 300 min. Urine and fecal samples were collected for 4 d after (45)Ca administration. Femurs were measured for bone mineral density (BMD), breaking strength, and total calcium. Calcium absorption, femoral calcium content, BMD, and bone balance (V(bal)) were significantly increased (P < 0.05) by NDO, whereas the bone resorption rate relative to the bone formation rate was significantly depressed by NDO. We conclude that feeding NDO at 5.5 g/100 g for 21 d has a positive effect on calcium absorption and retention in ovariectomized rats. PMID- 14747680 TI - The mode of oral bovine lactoferrin administration influences mucosal and systemic immune responses in mice. AB - Food protein intake interacts with the immune system. In earlier nutritional and immunological studies, nutrients, particularly milk whey proteins, were generally administered in soluble form and by gavage. However, orogastric intubation does not represent a natural way of ingesting nutrients such as lactoferrin (Lf). We examined how different modes of oral administration of Lf could affect the regulatory effect of this molecule on intestinal and systemic immune responses. Groups of 10 female BALB/c mice were administered Lf daily for 6 wk. To address the influence of the oral modes of administration, mice were given Lf either in solution, by gastric intubation or in the drinking water, or as a powder, by buccal deposition or in the diet. Mucosal and systemic immune responses, including specific immunoglobulin (Ig) secretion, cell proliferation, and cytokine production, were analyzed and compared with those of naive mice given water under the same conditions or positive control mice that were administered Lf by i.m. injection. The addition of Lf to the drinking water had no visible effect on the immune status. Gastric intubation, single buccal doses, and continuous doses of Lf in the diet stimulated transient systemic and intestinal antibody responses against Lf. All of these oral modes of Lf exposure biased mucosal and systemic T-cell responses toward Thelper (Th)2-types and elevated IgA production by mucosal cells. However, the less natural gastric intubation also promoted Th1-type responses as evidenced by serum IgG(2a) antibodies and the secretion of Th1 cytokine by mucosal and systemic T cells in vitro. Thus, one should carefully consider the oral mode of administration for understanding regulation of immune responses by food proteins such as Lf. PMID- 14747681 TI - Hamsters fed diets high in saturated fat have increased cholesterol accumulation and cytokine production in the aortic arch compared with cholesterol-fed hamsters with moderately elevated plasma non-HDL cholesterol concentrations. AB - There is growing evidence that dietary fatty acids and/or dietary cholesterol could have a direct role on inflammatory diseases such as atherosclerosis. F(1)B Golden Syrian hamsters (Mesocricetus auratus), in 2 groups of 72, were fed for 10 wk a semipurified diet containing either 20 g/100 g hydrogenated coconut oil without cholesterol or cocoa butter (20 g/100 g) with cholesterol (0.15 g/100 g). After the 10-wk treatment period, plasma was collected from food-deprived hamsters (16 h) for plasma lipid measurements. Hamsters were then ranked according to their plasma VLDL and LDL cholesterol (non-HDL-C) concentrations with 1.86 mmol/L as the cut-off point between low (Low; n = 36) and medium (Med; n = 36) concentrations for each treatment. Hamsters in the Low and Medium groups fed cholesterol (Low-chol) had significantly lower plasma total cholesterol (TC) concentrations than hamsters in the Low group fed coconut oil (Low-CO). However, this difference for the Medium group was reflected in significantly lower plasma HDL cholesterol (HDL-C) concentrations. Hamsters in the Low-CO group had significantly higher aortic total and esterified cholesterol concentrations than hamsters in the Low-chol group. Hamsters in the Low-chol group had significantly higher aortic tumor necrosis factor-alpha concentrations than hamsters in the Low CO group. Hamsters in the Med-CO group had significantly higher aortic interleukin-1beta concentrations than hamsters in the Med-chol group. In conclusion, the present study suggests that dietary cholesterol and saturated fatty acids could have an effect on atherosclerosis not only beyond their role in affecting plasma lipoproteins but also through increased production of inflammatory cytokines in the arterial wall. PMID- 14747682 TI - Contrasting and cooperative effects of copper and iron deficiencies in male rats fed different concentrations of manganese and different sources of sulfur amino acids in an AIN-93G-based diet. AB - Dietary nutrient interactions are important factors to consider in the study of nutrient status and requirements. Here, the effects of dietary interactions among copper (Cu), iron (Fe), manganese (Mn) and sulfur amino acids (SAA) on blood cell characteristics and enzyme activities were observed. Male rats (n = 8) were used in a 2 x 2 x 2 x 2 factorial design and fed an AIN-93G-based diet containing dietary Cu (<1 and 5 mg/kg), Fe (10 and 35 mg/kg), Mn (10 and 50 mg/kg) and either L-cystine (LCys) or DL-methionine (DLMet). Blood was analyzed by automated hematology cell counting and by flow cytometry. Severe Cu deficiency was verified by reductions in the activities of serum ceruloplasmin (1% of control), RBC superoxide dismutase (SOD1) (14% of control), liver cytochrome c oxidase activity (25% of control) and serum extracellular SOD (SOD3) activity (20% of controls). Because Cu is required for Fe utilization, many physiologic responses that require Fe were affected by both deficiencies, including lowered blood hemoglobin (Hgb), lower RBC volume and Hgb concentration, and an increased number of reticulocytes. Cu and Fe deficiencies together worsened some conditions, i.e., lower Hgb, lower RBC Hgb, increased RBC distribution width, increased number of reticulocytes and nucleated RBC, and a higher platelet count. Increasing dietary Mn had little effect on most variables, except to reduce serum Cu when dietary Cu was adequate but not when it was low, and to reduce RBC SOD1 activity when dietary Fe was low but not when it was adequate. Hgb concentrations were higher (P < 0.002) in Cu-deficient rats fed LCys than in those fed DLMet. There was no effect in Cu-adequate rats. Hgb was higher (P < 0.004) in Fe-adequate rats fed LCys than in those fed DLMet, with no effect in Fe-deficient rats. Although the anemia of Cu deficiency in AIN-93G-fed rats was not as pronounced as that reported in rats fed the AIN-76A-based diet, other manifestations of the deficiency were prominent. PMID- 14747683 TI - Alpha-tocopherol and ascorbic acid decrease the production of beta-apo-carotenals and increase the formation of retinoids from beta-carotene in the lung tissues of cigarette smoke-exposed ferrets in vitro. AB - Previously, we found that exposing ferrets to cigarette smoke enhanced oxidative excentric cleavage of beta-carotene. In the present study, we examined whether alpha-tocopherol, ascorbic acid, or the two combined can prevent smoke-altered beta-carotene metabolism. In vitro incubation of beta-carotene (10 micro mol/L) with lung postnuclear fractions from ferrets exposed to cigarette smoke was carried out in the absence or presence of alpha-tocopherol (50 micro mol/L), ascorbic acid (10 or 50 micro mol/L), or both vitamins to evaluate their effects on the production of beta-apo-carotenals and retinoids from beta-carotene. The oxidative cleavage metabolites of beta-carotene, beta-apo-carotenals (beta-apo 14', beta-apo-12', beta-apo-10', and beta-apo-8'), retinoic acid (RA), and retinal were analyzed by HPLC. We found that the smoke-enhanced production of individual beta-apo-carotenals was significantly decreased by 36-77% when alpha tocopherol (50 micro mol/L) and ascorbic acid (50 micro mol/L) were added together to the incubation mixture. alpha-Tocopherol alone had a modest effect. Ascorbic acid in the presence of alpha-tocopherol inhibited the production of beta-apo-carotenals in a dose-dependent manner, although ascorbic acid alone had no effect. In contrast, the production of RA and retinal among smoke-exposed ferrets was substantially increased ( approximately 3-fold, P < 0.05) when both alpha-tocopherol and ascorbic acid were added to the incubation mixtures. However, when ascorbic acid or alpha-tocopherol alone was added, the production of RA among smoke-exposed ferrets increased only modestly (80%, P < 0.05) and did not differ from the RA levels in control ferrets. In conclusion, these data indicate that alpha-tocopherol and ascorbic acid may act synergistically in preventing the enhanced oxidative excentric cleavage of beta-carotene induced by smoking exposure, thereby facilitating the conversion of beta-carotene into RA and retinal. PMID- 14747684 TI - Prima facie evidence that a phytocystatin for transgenic plant resistance to nematodes is not a toxic risk in the human diet. AB - A protein-engineered rice cystatin (OcIDeltaD86) provides transgenic, partial crop resistance to plant nematodes. This study determined whether its oral uptake has adverse effects on male Sprague-Dawley rats when they are administered by oral gavage 0.1-10 mg OcIDeltaD86/kg body weight daily for 28 d. Body weight and water and food intakes were unaltered for most of the study. The only significant changes in fresh weight of nine organs were for the liver (4% decrease; P < 0.05) and the empty cecum (14% increase; P < 0.05) at the two lowest doses and the highest dose of OcIDeltaD86, respectively. No abnormalities in either organ were detected by histochemistry. There were no changes in the urine or in hematological variables measured, and blood serum revealed no dose-dependent responses for any of 17 variables measured. OcIDeltaD86 was degraded by boiling with a 50% loss of its inhibition of papain after 9.2 +/- 8.0 min. It also showed >95% loss of such inhibition after 15 s in simulated gastric fluid. The results suggest that the no effect level (NOEL) for OcIDeltaD86 is >10 mg/(kg. d). This provides a range of dietary exposure >200-2000 fold depending upon the promoter used to control its expression in potato. PMID- 14747685 TI - Ascorbic acid does not increase the oxidative stress induced by dietary iron in C3H mice. AB - Iron is a potent prooxidant that can induce lipid peroxidation. Ascorbic acid, a potent antioxidant, has prooxidant effects in the presence of iron in vitro. We investigated whether ascorbic acid and iron co-supplementation in ascorbic acid sufficient mice increases hepatic oxidative stress. C3H/He mice were fed diets supplemented with iron to 100 mg/kg diet or 300 mg/kg diet with or without ascorbic acid (15 g/kg diet) for 3 wk. Liver iron concentration, malondialdehyde (MDA), glutathione (GSH), glutathione S-transferase (GST), glutathione peroxidase (GPx), superoxide dismutase (SOD), and catalase (CAT) were measured. High dietary iron increased liver iron concentrations slightly (P < 0.05), whereas it dramatically increased hepatic MDA (P < 0.0001). Ascorbic acid increased MDA but only in mice fed the low-iron diet (P < 0.05). The high-iron diet reduced GPx (P < 0.0001), CAT (P < 0.0005), SOD (P < 0.05), and GST (P < 0.005) activities regardless of ascorbic acid supplementation. In contrast, ascorbic acid reduced GPx (P < 0.0001) and CAT (P < 0.05) activities only in mice fed the low-iron diet. In conclusion, ascorbic acid supplementation can have prooxidant effects in the liver. However, ascorbic acid does not further increase the oxidative stress induced by increased dietary iron. PMID- 14747686 TI - Vitamin intake is not associated with community-acquired pneumonia in U.S. men. AB - The age-related decline in immune function may predispose individuals to increased infection risk. Nutrition is an important determinant of immunocompetence, but vitamin supplementation in relation to infection has not been evaluated extensively in well-nourished populations. We evaluated the associations between intakes of antioxidants and B vitamins and risk of community acquired pneumonia in well-nourished, middle-aged and older men. This was a prospective study conducted between 1990 and 2000 among 38,378 male, U.S. health professionals, aged 44 to 79 y in 1990. Participants answered a detailed 131-item FFQ to assess diet and also provided information on vitamin supplement use. We included those who at the onset had no history of pneumonia, myocardial infarction, stroke, other heart disease, arterial surgery, cancer or asthma, and also had complete dietary data. There were 446 new cases of nonfatal community acquired pneumonia during 145,878 person-years of follow-up. After adjustment for age, smoking, BMI, alcohol use, physical activity, diabetes and total energy intake, there were no associations between total intakes of antioxidants or B vitamins and pneumonia risk. After excluding men who took vitamin E supplements, vitamin E intake from food sources only was inversely associated with pneumonia risk (multivariate relative risk comparing extreme quintiles = 0.58, 95% CI, 0.39 0.86, P-value test for trend = 0.01). Vitamin supplements are unlikely to reduce pneumonia risk in well-nourished, middle-aged and older men. PMID- 14747687 TI - Urinary total flavonoid excretion but not 4-pyridoxic acid or potassium can be used as a biomarker for the intake of fruits and vegetables. AB - To gain better insight into the potential health effects of fruits and vegetables, reliable biomarkers of intake are needed. The main purpose of this study was to investigate the ability of flavonoid excretion in both 24-h and morning urine samples to reflect a low intake and moderate changes in fruit and vegetable consumption. Furthermore, the urinary excretions of 4-pyridoxic acid (4 PA) and potassium were investigated as other potential biomarkers of fruit and vegetable intake. The study was designed as a 5-d randomized, controlled crossover study. On d 1-3, the men (n = 12) consumed a self-restricted flavonoid free diet. On d 4, they were provided a strictly controlled diet containing no fruits or vegetables (basic diet). On d 5, they consumed the basic diet supplemented with 300 or 600 g of fruits and vegetables. The total excretion of flavonoids in 24-h urine samples increased linearly with increasing fruit and vegetable intakes (r(s) = 0.86, P < 1 x 10(-6)). The total excretion of flavonoids in morning urine also increased, but the association was weaker (r(s) = 0.59, P < 0.0001). Urinary 4-PA in 24-h and morning urine samples increased significantly only with the 600-g increase in fruit and vegetable intake, whereas the excretion of potassium in urine did not reflect the changes in fruit and vegetable intake. We conclude that the total excretion of flavonoids in 24-h urine may be used as a new biomarker for fruit and vegetable intake. PMID- 14747688 TI - Telephone counseling intervention increases intakes of micronutrient- and phytochemical-rich vegetables, fruit and fiber in breast cancer survivors. AB - Although a large body of evidence suggests that diet may play an important role in cancer prevention, randomized controlled trials reported to date have not achieved sufficient increases in protective micronutrients and phytochemicals to adequately test the hypothesis that diet can reduce cancer risk. The Women's Healthy Eating and Living (WHEL) Study, a randomized controlled trial of the role diet modification may play in future breast cancer events, introduced an innovative theory-based telephone counseling intervention to teach participants to consume a high fiber, low fat diet emphasizing vegetables and fruits rich in carotenoids and other potentially protective phytochemicals. This report examines the baseline to 12-mo changes in dietary intakes of 2970 participants, assessed through 24-h recalls and validated with plasma carotenoid concentrations. At 12 mo, the intervention group reported a significantly increased daily vegetable intake (+vegetable juice) of 7.1 servings (+82%) and fruit intake of 3.9 servings (+18%). Fiber intake increased from 3.04 to 4.16 g/(MJ. d), whereas energy from fat decreased significantly from 28.6 to 23.7%. Plasma carotenoid concentrations increased significantly, i.e., alpha-carotene (+223%); beta-carotene (+87%); lutein (+29%); and lycopene (+17%). In the comparison group, dietary intake and plasma carotenoid concentrations were essentially identical to those of the intervention group at baseline and were unchanged at 12 mo. The WHEL Study showed that a telephone counseling intervention can achieve major increases in micronutrient- and phytochemical-rich vegetables, fruit and fiber intakes, enabling an investigation of the potential cancer preventive effects of these food components. PMID- 14747689 TI - Colonic bacterial flora: changing understandings in the molecular age. AB - The human intestinal microbiota is a complex bacterial consortium that is critical to normal health. The microflora is present at concentrations of 10(11) 10(12) cells/g of intestinal contents; the number of species present may exceed 500, although exact numbers remain to be defined, due in part to the fact that <30% of microorganisms are culturable with current microbiologic methods. Molecular tools based on 16S rDNA sequence similarities such as fluorescent in situ hybridization (FISH), denaturing gradient gel electrophoresis (DGGE), quantitative dot blot hybridization, restriction fragment length polymorphism (RFLP) and large scale 16S rDNA sequencing have helped to overcome limitations of conventional microbiological plating methods in studying the fecal microflora composition. However, these tools are just now beginning to be applied to understand the dynamics of this complex community, and its relationship to diet and human health. There is a need to understand both the limitations of the current data and the importance of moving forward with the best possible molecular and epidemiologic techniques as we deal with these critical questions. PMID- 14747690 TI - Molecular ecological analysis of the gastrointestinal microbiota: a review. AB - The gastrointestinal (GI) microbiota of mammals is characterized by its high population density, wide diversity and complexity of interactions. While all major groups of microbes are represented, bacteria predominate. Importantly, bacterial cells outnumber animal (host) cells by a factor of ten and have a profound influence on nutritional, physiological and immunological processes in the host animal. Our knowledge of the molecular and cellular bases of host microbe interactions is limited, though critically needed to determine if and how the GI microbiota contributes to various enteric disorders in humans and animals. Traditionally, GI bacteria have been studied via cultivation-based techniques, which are labor intensive and require previous knowledge of individual nutritional and growth requirements. Recently, findings from culture-based methods have been supplemented with molecular ecology techniques that are based on the 16S rRNA gene. These techniques enable characterization and quantification of the microbiota, while also providing a classification scheme to predict phylogenetic relationships. The choice of a particular molecular-based approach depends on the questions being addressed. Clone libraries can be sequenced to identify the composition of the microbiota, often to the species level. Microbial community structure can be analyzed via fingerprinting techniques, while dot blot hybridization or fluorescent in situ hybridization can measure abundance of particular taxa. Emerging approaches, such as those based on functional genes and their expression and the combined use of stable isotopes and biomarkers, are being developed and optimized to study metabolic activities of groups or individual organisms in situ. Here, a critical summary is provided of current molecular ecological approaches for studying the GI microbiota. PMID- 14747691 TI - Effects of a controlled diet and black tea drinking on the fecal microflora composition and the fecal bile acid profile of human volunteers in a double blinded randomized feeding study. AB - Although diet has been clearly associated with human health many potential mechanisms remain undefined. For instance, although the intestinal bacterial microflora has long been postulated to contribute to human health, little is known about the effects of diet on the bacterial microflora composition and the specific contributions of the microflora to human health. Thus, we analyzed 1) changes in the fecal microflora composition by fluorescent in-situ hybridization (FISH) and denaturing gradient gel electrophoresis (DGGE) and 2) changes in the fecal bile acid profile, in a crossover feeding study that investigated the effects of black tea drinking on blood lipids in hypercholesterolemic volunteers. DGGE analysis shows that each study subject harbors a specific bacterial profile that exhibits little change over time. Change from a "free" living diet to the controlled study diet or to black tea drinking did not significantly change these bacterial profiles. FISH analysis revealed that even though black tea did not affect the specific bacterial groups that were analyzed, it did decrease the amounts of bacteria that were detected by the universal bacterial probe, but not by any of the specific probes. We did not detect any consistent effects of either diet or black tea drinking on the levels and proportions of fecal bile acids. Our results indicate that tea drinking affects some microflora components. Larger studies with well defined end points that control for the observed variation are needed to improve our understanding of the effects of diet on intestinal microflora and fecal bile acid profile. PMID- 14747692 TI - Microbial degradation products influence colon cancer risk: the butyrate controversy. AB - All dietary fiber, by definition, escapes digestion in the small intestine and thus arrives relatively intact in the large intestine. Its fate in the large intestine depends upon the type of fiber and the colonic microflora. Highly fermentable fibers result in short chain fatty acids including butyrate, which is thought by some to be protective against colon cancer. However, not all studies support a chemopreventive effect for butyrate and the lack of agreement (particularly between in vivo and in vitro studies) on butyrate and colon cancer has been termed the "butyrate paradox." There are a number of reasons for this discrepant effect including differences between the in vitro and in vivo environments, the timing of butyrate administration, the amount of butyrate administered, the source of butyrate (usually dietary fiber) as a potential confounder, and an interaction with dietary fat. Collectively, the studies suggest that the chemopreventive benefits of butyrate depend in part on amount, time of exposure with respect to the tumorigenic process, and the type of fat in the diet. PMID- 14747693 TI - Ursodeoxycholic acid (UDCA) can inhibit deoxycholic acid (DCA)-induced apoptosis via modulation of EGFR/Raf-1/ERK signaling in human colon cancer cells. AB - Ursodeoxycholic acid (UDCA), a hydrophilic bile acid, is known as a cytoprotective agent. UDCA prevents apoptosis induced by a variety of stress stimuli including cytotoxic bile acids such as deoxycholic acid (DCA). Here we examined the molecular mechanism by which UDCA can antagonize DCA-induced apoptosis in human colon cancer cells. UDCA pretreatment decreases the number of apoptotic cells caused by exposure to DCA and UDCA. Further studies of the signaling pathway showed that UDCA pretreatment suppressed DNA binding activity of activator protein-1 and this was accompanied by downregulation of both extracellular signal-regulated kinase (ERK) and Raf-1 kinase activities stimulated by exposure to DCA. DCA was also found to activate epidermal growth factor receptor (EGFR) activity and UDCA inhibited this. Collectively, these findings suggest that the inhibitory effect of UDCA in DCA-induced apoptosis is partly mediated by modulation of EGFR/Raf-1/ERK signaling. PMID- 14747694 TI - HUPO initiatives relevant to clinical proteomics. AB - The past few years have seen a tremendous interest in the potential of proteomics to address unmet needs in biomedicine. Such unmet needs include more effective strategies for early disease detection and monitoring and more effective therapies, in addition to developing a better understanding of disease pathogenesis. Proteomics is particularly suited for investigating biological fluids to identify disease-related alterations and to develop molecular signatures for disease processes. However, much of the effort undertaken in clinical proteomics to date represents either demonstrations of principles or relatively small-scale studies when compared with genomics effort and accomplishments or more pertinently when contrasted with the tremendous untapped potential of clinical proteomics. Clearly, we are in the early stages. What seems to be urgently needed is an organized effort to build a solid foundation for proteomics that includes developing a much needed infrastructure with adequate resources. The Human Proteome Organization (HUPO) is fostering an organized international effort in proteomics that includes initiatives around organ systems and biological fluids that have disease relevance as well as development of proteomics resources. PMID- 14747695 TI - Nuclear receptors: the evolution of diversity. AB - Nuclear receptors are an ancient family of transcription factors. Some receptors are regulated by small lipophilic ligands, whereas others are constitutive transcriptional activators or repressors. The evolution of this diversity is poorly understood, and it remains an open question as to whether or not the ancestral receptor was ligand-regulated. The recent cloning, from a snail, of an estrogen receptor that does not bind estrogen not only suggests that the steroid receptors are much more ancient than previous thought, but also points toward a mechanism through which nuclear receptors can lose the ability to be ligand regulated. PMID- 14747696 TI - Structural plasticity of peanut lectin: an X-ray analysis involving variation in pH, ligand binding and crystal structure. AB - Until recently, it has only been possible to grow crystals of peanut lectin when complexed with sugar ligands. It is now shown that it is possible to grow peanut lectin crystals at acidic pH in the presence of oligopeptides corresponding to a loop in the lectin molecule. Crystals have also been prepared in the presence of these peptides as well as lactose. Low-pH crystal forms of the lectin-lactose complex similar to those obtained at neutral pH have also been grown. Thus, crystals of peanut lectin grown under different environmental conditions, at two pH values with and without sugar bound to the lectin, are now available. They have been used to explore the plasticity and hydration of the molecule. A detailed comparison between different structures shows that the lectin molecule is sturdy and that the effect of changes in pH, ligand binding and environment on it is small. The region involving the curved front beta-sheet and the loops around the second hydrophobic core is comparatively rigid. The back beta-sheet involved in quaternary association, which exhibits considerable variability, is substantially flexible, as is the sugar-binding region. The numbers of invariant water molecules in the hydration shell are small and they are mainly involved in metal coordination or in stabilizing unusual structural features. Small consistent movements occur in the combining site upon sugar binding, although the site is essentially preformed. PMID- 14747697 TI - The influence of positional errors on the Debye effects. AB - The relation between a Gaussian perturbation of the atomic positional parameters and the average squared structure-factor amplitude is presented. Using an error dependent radial distance distribution of an atomic protein model, it can be shown that the Debye effects diminish exponentially as a function of increasing positional errors. These relations can be used to estimate the quality of an atomic model and the corresponding phases. The limiting case of equal atoms with an infinitely large coordinate error results in the classical Wilson model. PMID- 14747698 TI - On the interpretation and use of <|E|2>(d*) profiles. AB - Profiles of squared normalized structure factors, <|E|(2)>(d*), have been computed for a large number of proteins and nucleic acids. These are interpreted in terms of their underlying structural features. It is also shown that the 'solvent dip' at around 6.3 A resolution is to a large extent a protein secondary structure effect that is enhanced by the water structure. A hierarchical classification of protein structures based on their <|E|(2)>(d*) profiles is briefly outlined, together with the use of <|E|(2)>(d*) profiles as an improvement over Wilson absolute scale estimation and as a novel solvent modelling method. PMID- 14747699 TI - A preliminary time-of-flight neutron diffraction study of Streptomyces rubiginosus D-xylose isomerase. AB - The metalloenzyme D-xylose isomerase forms well ordered crystals that diffract X rays to ultrahigh resolution (<1 A). However, structural analysis using X-ray diffraction data has as yet been unable to differentiate between several postulated mechanisms that describe the catalytic activity of this enzyme. Neutrons, with their greater scattering sensitivity to H atoms, could help to resolve this by determining the protonation states within the active site of the enzyme. As the first step in the process of investigating the mechanism of action of D-xylose isomerase from Streptomyces rubiginosus using neutron diffraction, data to better than 2.0 A were measured from the unliganded protein at the Los Alamos Neutron Science Center Protein Crystallography Station. Measurement of these neutron diffraction data represents several milestones: this is one of the largest biological molecules (a tetramer, MW approximately 160 000 Da, with unit cell lengths around 100 A) ever studied at high resolution using neutron diffraction. It is also one of the first proteins to be studied using time-of flight techniques. The success of the initial diffraction experiments with D xylose isomerase demonstrate the power of spallation neutrons for protein crystallography and should provide further impetus for neutron diffraction studies of biologically active and significant proteins. Further data will be measured from the enzyme with bound substrates and inhibitors in order to provide the specific information needed to clarify the catalytic mechanism of this enzyme. PMID- 14747700 TI - Solving the structure of the bubble protein using the anomalous sulfur signal from single-crystal in-house Cu Kalpha diffraction data only. AB - A small cysteine-rich protein, the function of which remains elusive, was discovered in the exudate of a Penicillium species. Crystal diffraction experiments conducted using in-house Cu Kalpha radiation and an R-AXIS IV++ imaging-plate detector yielded high-quality data to 1.4 A, with a distinguishable anomalous signal from sulfur (DeltaF/F = 0.031). This was used to phase the data and solve the structure using a single data set; the 64-residue amino-acid sequence was unambiguously determined from the electron density. It revealed a globular all-beta protein with a hitherto unknown fold, having a surface electrostatic charge distribution that is similar to that of another small secreted fungal protein, the Williopsis mrakii killer toxin. Aligning the charge distribution superimposed the potential recognition sites of the two proteins, suggesting a similar negatively charged target. PMID- 14747701 TI - The structure of a universally employed enzyme: V8 protease from Staphylococcus aureus. AB - V8 protease, an extracellular protease of Staphylococcus aureus, is related to the pancreatic serine proteases. The enzyme cleaves peptide bonds exclusively on the carbonyl side of aspartate and glutamate residues. Unlike the pancreatic serine proteases, V8 protease possesses no disulfide bridges. This is a major evolutionary difference, as all pancreatic proteases have at least two disulfide bridges. The structure of V8 protease shows structural similarity with several other serine proteases, specifically the epidermolytic toxins A and B from S. aureus and trypsin, in which the conformation of the active site is almost identical. V8 protease is also unique in that the positively charged N-terminus is involved in determining the substrate-specificity of the enzyme. PMID- 14747702 TI - On the possibility of the observation of valence electron density for individual bonds in proteins in conventional difference maps. AB - In the last decade, high-resolution data have become available for macromolecular objects. Furthermore, ultrahigh-resolution diffraction data (resolution close to 0.6 A) have been collected for several protein crystals. This allows the study of fine details of the electron-density distribution such as the deformation density, i.e. the deviation of the experimentally determined electron density from the density composed of 'free' non-bonded atoms. This paper discusses the resolution and atomic temperature factors necessary to make the valence electron density visible at individual bonds in conventional difference maps for macromolecules. The study of theoretical maps calculated by quantum-chemistry methods allows estimation of these conditions; these results are confirmed by analysis of experimental maps for Leu-enkephalin and antifreeze protein RD1. A resolution limit close to 0.6 A was found to be highly important for refinement even when the maps were calculated at lower resolution. The refinement of the same models at near to 0.9 A resolution results in artificially increased values of the atomic displacement parameters and does not permit bond electron density to be visible in difference maps. To some extent, overestimation of the atomic displacement parameters may be restricted if dummy bond electrons are used in the refinement. PMID- 14747703 TI - The impact of Lys-->Arg surface mutations on the crystallization of the globular domain of RhoGDI. AB - The potential of rational surface mutagenesis for enhanced protein crystallization is being probed in an ongoing effort. In previous work, it was hypothesized that residues with high conformational entropy such as Glu and Lys are suitable targets for surface mutagenesis, as they are rarely incorporated in crystal contacts or protein-protein interfaces. Previous experiments using Lys- >Ala, Glu-->Ala and Glu-->Asp mutants confirmed that mutated proteins were more likely to crystallize. In the present paper, the usefulness of Lys-->Arg mutations is studied. Several mutations of the globular domain of human RhoGDI were generated, including the single mutants K105R, K113R, K127R, K138R and K141R, the double mutants K(98,99)R and K(199,200)R and the triple mutants K(98,99,105)R and K(135,138,141)R. It is shown that Lys-->Arg mutants are more likely to crystallize than the wild-type protein, although not as likely as Lys- >Ala mutants. Out of the nine mutants tested, five produced diffracting crystals, including the K(199,200)R double mutant, which crystallized in a new space group and exceeded by approximately 1.0 A the resolution of the diffraction of the wild type crystal. Major crystal contacts in the new lattice were created by the mutated epitope. PMID- 14747704 TI - Porphyrin binding to jacalin is facilitated by the inherent plasticity of the carbohydrate-binding site: novel mode of lectin-ligand interaction. AB - The crystal structure of the complex of meso-tetrasulfonatophenylporphyrin (H(2)TPPS) with jack fruit (Artocarpus integriflora) agglutinin (jacalin) has been determined at 1.8 A resolution. A porphyrin pair is sandwiched between two symmetry-related jacalin monomers in the crystal, leading to a cross-linking network of protein molecules. Apart from the stacking interactions, H(2)TPPS also forms hydrogen bonds, some involving water bridges, with jacalin at the carbohydrate-binding site. The residues that are involved in rendering galactopyranoside specificity to jacalin undergo conformational adjustments in order to accommodate the H(2)TPPS molecule. The water molecules at the carbohydrate-binding site of jacalin cement the jacalin-porphyrin interactions, optimizing their complementarity. Interactions of porphyrin with jacalin are relatively weak compared with those observed between galactopyranoside and jacalin, perhaps because the former largely involves water-mediated hydrogen bonds. While H(2)TPPS binds to jacalin at the carbohydrate-binding site as in the case of ConA, its mode of interaction with jacalin is very different. H(2)TPPS does not enter the carbohydrate-binding cavity of jacalin. Instead, it sits over the binding site. While the porphyrin binding is mediated by replicating the hydrogen-bonding network of mannopyranoside through the sulfonate atoms in the case of ConA, the plasticity associated with the carbohydrate-binding site accommodates the pluripotent porphyrin molecule in the case of jacalin through an entirely different set of interactions. PMID- 14747705 TI - Structure of the core region of the soybean beta-conglycinin alpha' subunit. AB - The crystal structure of the core region of the alpha' subunit (alpha(c')) of soybean beta-conglycinin has been determined at 2.3 A resolution. alpha(c') was superimposed on the known crystal structure of the beta-conglycinin beta subunit with a small root-mean square deviation of 0.77 A, which is consistent with the high sequence identity of 75.5% between alpha(c') and the beta subunit. It is known that the thermal stability of the beta subunit is higher than that of the alpha' subunit and that their thermal stabilities are conferred by highly homologous core regions. Comparisons of the three-dimensional structures and primary sequences between alpha(c') and the beta subunit suggest that five factors account for this difference between subunits as regards the difference in thermal stability: (i) the total cavity volume is larger in alpha(c'), (ii) the cluster of charged residues at the intermonomer interface is smaller in alpha(c') and alpha(c') lacks the intermonomer salt bridge of the beta subunit, (iii) the solvent-accessible surface is more hydrophobic in alpha(c'), (iv) there are fewer proline residues in alpha(c') and (v) a loop region between helix 3 and strand J' in alpha(c') is more flexible owing to the insertion of five additional residues. Although more hydrogen bonds were found in alpha(c'), this difference should be more than compensated for by the combined contributions of these other factors. PMID- 14747706 TI - Metal-substituted derivatives of the rubredoxin from Clostridium pasteurianum. AB - Five different metal-substituted forms of Clostridium pasteurianum rubredoxin have been prepared and crystallized. The single Fe atom present in the Fe(S Cys)(4) site of the native form of the protein was exchanged in turn for Co, Ni, Ga, Cd and Hg. All five forms of rubredoxin crystallized in space group R3 and were isomorphous with the native protein. The Co-, Ni- and Ga-substituted proteins exhibited metal sites with geometries similar to that of the Fe form (effective D(2d) local symmetry), as did the Cd and Hg proteins, but with a significant expansion of the metal-sulfur bond lengths. A knowledge of these structures contributes to a molecular understanding of the function of this simple iron-sulfur electron-transport protein. PMID- 14747707 TI - Structure of circularly permuted DsbA(Q100T99): preserved global fold and local structural adjustments. AB - The thiol-disulfide oxidoreductase DsbA is required for efficient formation of disulfide bonds in the Escherichia coli periplasm. The enzyme is the strongest oxidant of the family of thioredoxin-like proteins and three-dimensional structures of both oxidized and reduced forms are known. DsbA consists of a catalytic thioredoxin-like domain and a helical domain that is inserted into the thioredoxin motif. Here, the X-ray structure of a circularly permuted variant, cpDsbA(Q100T99), is reported in which the natural termini are joined by the pentapeptide linker GGGTG, leading to a continuous thioredoxin domain, and new termini that have been introduced in the helical domain by breaking the peptide bond Thr99-Gln100. cpDsbA(Q100T99) is catalytically active in vivo and in vitro. The crystal structure of oxidized cpDsbA(Q100T99), determined by molecular replacement at 2.4 A resolution, was found to be very similar to that of wild type DsbA. The lower thermodynamic stability of cpDsbA(Q100T99) relative to DsbA is associated with small structural changes within the molecule, especially near the new termini and the circularizing linker. The active-site helices and adjacent loops display increased flexibility compared with oxidized DsbA. PMID- 14747708 TI - Crystallization and preliminary crystallographic data of a leucotoxin S component from Staphylococcus aureus. AB - Class S proteins of staphylococcal bicomponent pore-forming leucotoxins play an important role in membrane targetting and cell specificity. Wild-type and recombinant S components of the Panton-Valentine leucocidin (LukS-PV) were expressed in Staphylococcus aureus and Escherichia coli, respectively, and purified. Both proteins were crystallized in two crystal forms with Jeffamine M 600 as the precipitant at 285 K using the hanging-drop vapour-diffusion method and seeding techniques. Crystals belong to space group P2 (or P2(1)) and P4(1) (or P4(3)), with unit-cell parameters a = 72.3, b = 95.1, c = 108.1 A, beta = 106.4 degrees and a = b = 94.8, c = 306.2 A, respectively. A full set of X-ray diffraction data was collected to 2.1 A from a single tetragonal crystal of the wild-type protein at 100 K. PMID- 14747709 TI - Purification, crystallization and preliminary X-ray diffraction of a proteolytic fragment of PDK1 containing the pleckstrin homology domain. AB - 3-Phosphoinositide-dependent protein kinase-1 (PDK1) is a Ser/Thr kinase with an essential role in insulin and growth-factor signalling. PDK1 activity towards protein kinase B (PKB) is partially regulated by its pleckstrin homology (PH) domain, which preferentially binds to 3-phosphoinositides. However, the precise molecular mechanism of this regulation is not well understood. Here, the cloning, purification and crystallization of a 150-amino-acid C-terminal region of PDK1 containing the PH domain is reported. A crystal of the PDK1 PH domain grown in the presence of inositol 1,3,4,5-tetrakisphosphate and derivatized with AuCN diffracted to 1.5 A at a synchrotron source. Diffraction data collected near the Au edge resulted in an anomalous Patterson map with a 30sigma peak. PMID- 14747710 TI - Crystallization and preliminary crystallographic analysis of rat monoamine oxidase A complexed with clorgyline. AB - Monoamine oxidase (MAO) is an FAD-containing mitochondrial outer-membrane protein which catalyzes the degradation of several neurotransmitters in the central nervous system. The two subtypes of MAO, MAOA and MAOB, have similar primary sequences but different substrate and inhibitor specificities. The structure of human MAOB has recently been determined, but the structure of MAOA remains unknown. To clarify the mechanisms underlying their unique substrate and inhibitor recognition and thereby facilitate the development of new specific inhibitors to treat MAO-related neurological disorders, rat MAOA was crystallized in a complex with the specific inhibitor clorgyline. Diffraction data were collected to 3.2 A resolution. The crystal belongs to the space group P4(3)2(1)2, with unit-cell parameters a = b = 158.2, c = 258.4 A. PMID- 14747711 TI - Crystallization and preliminary X-ray analysis of the helicase domains of Vasa complexed with RNA and an ATP analogue. AB - The helicase fragment of Vasa was purified and its RNA-binding activity was examined by a UV cross-linking assay. The fragment was crystallized in complex with poly(U) RNA (U(10)) and a non-hydrolyzable analogue of ATP. The crystal belonged to space group P2(1), with unit-cell parameters a = 71.06, b = 142.35, c = 130.47 A, beta = 90.86 degrees. The cryocooled crystal diffracted to about 2.2 A using synchrotron radiation from station BL41XU at SPring-8. PMID- 14747712 TI - Crystallization and preliminary X-ray analysis of the controller protein C.AhdI from Aeromonas hydrophilia. AB - Single crystals of purified homodimeric controller protein from Aeromonas hydrophilia (C.AhdI) have been grown under several different conditions using vapour diffusion. X-ray diffraction data have been collected using synchrotron radiation from crystals of both the native and a selenomethionine (SeMet) derivative of the protein. The native crystal form belongs to space group P2(1) and data were collected to a resolution of 2.2 A. Two crystal forms of the SeMet protein have been obtained and were found to belong to space groups P1 and P2(1); data have been recorded to 2.0 and 1.7 A resolution, respectively, for the two crystal forms. Three-wavelength MAD data were collected to 1.7 A for the SeMet derivative crystal, which is isomorphous with the native P2(1) crystal. PMID- 14747713 TI - Expression, purification and crystallization of an extended-spectrum beta lactamase from Klebsiella oxytoca. AB - OXY-1a is an extended-spectrum beta-lactamase from the conditional pathogenic bacterium Klebsiella oxytoca. OXY-1a is responsible for the antibiotic resistance of this pathogen. A soluble form of OXY-1a with a His tag at its C-terminus was overexpressed in Escherichia coli. The recombinant protein was purified and crystallized at room temperature using PEG 4000 as the main precipitant. Two crystal forms were obtained from the same growth conditions. One was orthorhombic, with crystals that diffracted to better than 1.9 A, while the other was tetragonal, with crystals that only diffracted to about 3.0 A. Complete data sets were collected from both crystal forms. The orthorhombic crystal belongs to space group P2(1)2(1)2(1), with unit-cell parameters a = 46.54, b = 73.43, c = 84.56 A, while the tetragonal crystal has unit-cell parameters a = b = 73.72, c = 96.81 A. The asymmetric unit of the orthorhombic crystal is estimated to contain one OXY-1a molecule, giving a crystal volume per protein weight (V(M)) of 2.25 A(3) Da(-1) and a solvent content of 45%. PMID- 14747714 TI - Crystallization of YloQ, a GTPase of unknown function essential for Bacillus subtilis viability. AB - YloQ is a putative ATP/GTP-binding protein of unknown function identified from the complete sequence of the Bacillus subtilis genome. A gene-knockout programme established that yloQ is one of a set of some 270 indispensable genes for the viability of this organism. Crystals of YloQ have been grown from HEPES-buffered solutions at pH 7.5 containing polyethylene glycol and diffraction data have been collected extending to 2.5 A spacing. PMID- 14747715 TI - Crystallization and preliminary X-ray diffraction analysis of a dihaem cytochrome c peroxidase from Paracoccus denitrificans. AB - Cytochrome c peroxidase was isolated from Paracoccus denitrificans and purified to homogeneity in three steps prior to crystallization. Two different diffraction quality crystal forms were obtained by the hanging-drop vapour-diffusion method using a number of screening conditions. The best (needle-shaped) crystal form is suitable for structural studies and was grown from solutions containing 20% PEG 8000, 0.1 M Tris pH 8.5 and 0.2 M MgCl(2). Crystals grew to a maximum length of approximately 0.7 mm and belong to the primitive monoclinic space group P2(1), with unit-cell parameters a = 78.3, b = 51.0, c = 167.2 A, beta = 97.9 degrees. After a dehydration step and extensive optimization of the cryocooling conditions, a complete data set was collected to 2.2 A from a native crystal of the fully oxidized form of the enzyme using synchrotron radiation. PMID- 14747716 TI - Expression, crystallization and preliminary X-ray studies of the recombinant PTB domain of mouse dok1 protein. AB - The PTB domain of mouse dok1 fusion protein has been overexpressed in Escherichia coli and crystallized in a form suitable for X-ray crystallographic study. Crystals have been obtained using the vapour-diffusion method and belong to space group P2(1)2(1)2(1). X-ray diffraction data were collected in-house to 2.5 A resolution. A selenomethionine (SeMet) dok1 PTB fusion-protein derivative was expressed using the same expression system, purified in a reductive environment and crystals were obtained under similar conditions. Subsequently, three different wavelength data sets from the derivative crystal were collected to 2.5 A resolution at SPring-8. PMID- 14747717 TI - Crystallization and preliminary X-ray diffraction analysis of an oxidized state of Ohr from Xylella fastidiosa. AB - Xylella fastidiosa organic hydroperoxide-resistance protein (Ohr) is a dithiol dependent peroxidase that is widely conserved in several pathogenic bacteria with high affinity for organic hydroperoxides. The protein was crystallized using the hanging-drop vapour-diffusion method in the presence of PEG 4000 as precipitant after treatment with organic peroxide (t-butyl hydroperoxide). X-ray diffraction data were collected to a maximum resolution of 1.8 A using a synchrotron radiation source. The crystal belongs to the hexagonal space group P6(5)22, with unit-cell parameters a = b = 87.66, c = 160.28 A. The crystal structure was solved by molecular-replacement methods. The enzyme has a homodimeric quaternary structure similar to that observed for its homologue from Pseudomonas aeruginosa, but differs from the previous structure as the active-site residue Cys61 is oxidized. Structure refinement is in progress. PMID- 14747718 TI - Crystallization and preliminary crystallographic analysis of a novel orange fluorescent protein from the Cnidaria tube anemone Cerianthus sp. AB - A novel orange fluorescent protein, with excitation and emission maxima at 548 and 565 nm, respectively, from the Cnidaria tube anemone Cerianthus sp. has been cloned and overexpressed in Escherichia coli. The orange fluorescent protein has been crystallized by the sitting-drop vapour-diffusion method at 290 K using polyethylene glycol 3350 as a precipitant. A complete set of diffraction data was collected to 2.0 A resolution at 100 K. The crystals belong to the space group R3, with hexagonal unit-cell parameters a = b = 216.947, c = 51.839 A. There are four protein molecules in the asymmetric unit, giving a Matthews coefficient of 2.3 A(3) Da(-1) and a solvent content of 47%. PMID- 14747719 TI - Crystallization and preliminary X-ray analysis of xylanase B from Clostridium stercorarium. AB - Recombinant mature xylanase B from Clostridium stercorarium has been prepared and crystallized by the sitting-drop vapour-diffusion method using 4 mg ml(-1) purified enzyme, 10.3%(w/v) polyethylene glycol 1500, 8.6%(v/v) glycerol and 0.34 M non-detergent sulfobetaine 195. A suitable crystal grew after incubation for ten weeks at 293 K. The crystal belonged to space group P2(1)2(1)2(1), with unit cell parameters a = 64.76, b = 96.60, c = 138.44 A. X-ray diffraction data were collected to 1.80 A resolution. PMID- 14747720 TI - Cloning, purification, crystallization and preliminary crystallographic studies of Bradyrhizobium fucosyltransferase NodZ. AB - The alpha-1,6-fucosyltransferase NodZ from Bradyrhizobium sp. WM9 (Lupinus), composed of 325 amino acids with a molecular weight of 37 kDa, has been cloned, expressed and purified. Protein crystals suitable for X-ray diffraction were obtained under optimized crystallization conditions using ammonium dihydrogen phosphate as a precipitant. The crystals are hexagonal and belong to space group P6(1)22 or P6(5)22, with unit-cell parameters a = 125.5, c = 95.6 A, and contain 56.8% solvent and a single protein molecule in the asymmetric unit. Native data were collected to 2.85 A using synchrotron radiation and cryogenic conditions. The native crystals were soaked in a mother-liquor solution containing 2.5 mM [Ta(6)Br(12)](2+) cluster for derivatization and SAD data were collected to 3.4 A at the tantalum L(III) absorption peak. PMID- 14747721 TI - Crystallization and preliminary X-ray analysis of the human-specific toxin intermedilysin. AB - Intermedilysin is a human-specific toxin from Streptococcus intermedius, which is part of normal human oral flora. The bacterium is an opportunistic pathogen with a tendency for deep-seated infection in the brain and liver. Intermedilysin belongs to the cholesterol-dependent cytolysin (CDCs) family of toxins, which have been identified in several different bacteria including the serious human pathogens S. pneumoniae and Clostridium perfringens. Intermedilysin, however, is the only member that shows exclusive specificity for human cells. The toxin has a couple of non-conservative amino-acid substitutions in a tryptophan-rich region of the molecule (Cys to Ala and Trp to Pro), the most conserved region amongst the CDCs. Mutations in this region are known to render other CDCs inactive. In order to investigate the structure-function relationships of the unusual features of intermedilysin, which will help us to understand the molecular mechanism of the toxin family in general, recombinant intermedilysin has been crystallized. The crystals belong to an orthorhombic space group and contain two molecules per asymmetric unit. Diffraction data were collected to 2.3 A using synchrotron radiation. PMID- 14747722 TI - Expression, purification and crystallization of human 3'-phosphoadenosine-5' phosphosulfate synthetase 1. AB - 3'-Phosphoadenosine-5'-phosphosulfate (PAPS) is used to incorporate sulfate into biomolecules. The human PAPS synthetase 1 catalyzes two steps leading from adenosine triphosphate (ATP) and sulfate to PAPS. The ATP sulfurylase domain catalyzes the formation of the intermediate adenosine-5'-phosphosulfate (APS). The APS kinase domain then adds a phosphate group to the 3'-ribose and releases PAPS. In this article, the recombinant expression, purification and crystallization of the full-length protein is described. In Escherichia coli the protein is only partly soluble and copurifies with GroEL. The pure protein migrates as a dimer in gel-filtration chromatography. It is moderately active, forming 25 nmol PAPS per minute per milligram. Crystals grow to 100 x 100 x 300 micro m and diffract to 1.75 A. PMID- 14747724 TI - Expression, purification, crystallization and preliminary crystallographic analysis of osmotically inducible protein C. AB - Selenium-incorporated osmotically inducible protein C from the thermophilic bacterium Thermus thermophilus was overexpressed, purified and crystallized. The crystals belong to space group P1, with unit-cell parameters a = 37.58, b = 40.95, c = 48.14 A, alpha = 76.93, beta = 74.04, gamma = 64.05 degrees. Five data sets were collected from a single crystal to 1.6 A using synchrotron radiation for MAD phasing. Self-rotation functions and the Matthews coefficient are consistent with two molecules in the asymmetric unit. PMID- 14747723 TI - Crystallization and preliminary crystallographic analysis of Mycoplasma arthritidis-derived mitogen complexed with peptide/MHC class II antigen. AB - Mycoplasma arthritidis-derived mitogen (MAM), a bacterial superantigen, has been crystallized in complex with its human receptor, major histocompatibility complex (MHC) class II antigen, by the hanging-drop vapor-diffusion method. Crystals were obtained under three conditions, with ammonium sulfate, phosphate salt and PEG 8000 as the precipitant. The crystals grown under these conditions all belong to space group I222, with the same unit-cell parameters: a = 137.4, b = 178.2, c = 179.6 A. Diffraction data were collected to 3.3 and 3.4 A resolution from crystals of native and selenomethionylated MAM-MHC complexes, respectively. Self- and cross-rotation function calculations suggest the presence of two complex molecules in the asymmetric unit, resulting in a V(M) of 4.0 and a solvent content of 69%. An interpretable electron-density map was produced using a combination of molecular replacement and SAD phasing. PMID- 14747725 TI - Expression, purification, crystallization and preliminary characterization of uridine 5'-diphospho-N-acetylmuramoyl L-alanyl-D-glutamate:lysine ligase (MurE) from Streptococcus pneumoniae 110K/70. AB - An ORF designated sp1530 (murE) in the Streptococcus pneumoniae TIGR4 genome sequence, identified as uridine 5'-diphospho-N-acetylmuramoyl-L-alanyl-D glutamate:L-lysine ligase (MurE; EC 6.3.2.7), was cloned into the high-expression plasmid pET21b and overexpressed in Escherichia coli BL21 (DE3) Star. The enzyme was purified in three steps to 99% purity. Crystals were obtained by the hanging drop vapour-diffusion method at 291 K from solutions containing 25%(w/v) polyethylene glycol 2000 monomethylether, 0.2 M potassium thiocyanate, 0.1 M MES pH 6.5 in the presence of uridine 5'-diphospho-N-acetylmuramoyl alanyl glutamate (UDP-MurNAc-L-Ala-D-Glu) with and without 5'-adenylyl imidophosphate (AMP-PNP), a non-hydrolysable analogue of ATP. Diffraction data to 1.5 and 2.7 A, respectively, were collected at the European Synchrotron Radiation Facility (ESRF). Crystals grown in the presence of two ligands belong to space group P1, with unit-cell parameters a = 68.4, b = 71.4, c = 74.8 A, alpha = 73.4, beta = 80.5, gamma = 72.3 degrees. Crystals grown in the presence of UDP-MurNAc-L-Ala-D Glu alone belong to space group P2(1), with unit-cell parameters a = 71.1, b = 129.4, c = 74.6 A, beta = 106.3 degrees. PMID- 14747726 TI - Purification, crystallization and preliminary X-ray studies of sylvaticin, an elicitin-like protein from Pythium sylvaticum. AB - Sylvaticin belongs to the elicitin family. These 10 kDa oomycetous proteins induce a hypersensitive response in plants, including necrosis and cell death, but subsequently leading to a non-specific systemic acquired resistance (SAR) against other pathogens. Sylvaticin has been crystallized using PEG 2000 MME as a precipitant agent in the presence of nickel chloride. The crystals belong to space group C2, with unit-cell parameters a = 99.29, b = 25.67, c = 67.45 A, beta = 99.66 degrees. Diffraction data were recorded to 2.1 A resolution at a synchrotron-radiation source. PMID- 14747727 TI - Crystallization and preliminary X-ray crystallographic analysis of a ytfG gene product from Escherichia coli. AB - The Escherichia coli ytfG gene product, with NAD(P)H:quinone oxidoreductase activity, was crystallized by the hanging-drop vapour-diffusion method at 296 K. A 1.78 A data set has been collected using synchrotron radiation at Pohang Light Source, South Korea. The crystal belongs to the primitive trigonal system, with unit-cell parameters a = b = 81.7, c = 76.8 A. Analysis of the packing density shows that the asymmetric unit probably contains one monomer, with a solvent content of 48.8%. PMID- 14747728 TI - Purification, crystallization and preliminary X-ray studies of Mycobacterium tuberculosis RRF. AB - The ribosome recycling factor from Mycobacterium tuberculosis has been crystallized. The monoclinic crystals, with 52.5% solvent content, contain one protein molecule in the asymmetric unit. PMID- 14747729 TI - Purification, crystallization and preliminary X-ray diffraction analysis of the catalytic domain of adenylyl cyclase Rv1625c from Mycobacterium tuberculosis. AB - The Rv1625c gene product is an adenylyl cyclase identified in the genome of Mycobacterium tuberculosis strain H37Rv. It shows sequence similarity to the mammalian nucleotide cyclases and functions as a homodimer, with two substrate binding sites at the dimer interface. A mutant form of the catalytic domain of this enzyme, K296E/F363R/D365C (KFD-->ERC), was overexpressed in Escherichia coli cells in a soluble form. Crystals were obtained using the hanging-drop vapour diffusion method with PEG 8000 as a precipitant. The protein crystallized in space group P4(1), with unit-cell parameters a = b = 71.25, c = 44.51 A. X-ray diffraction data were collected to a resolution of 3.4 A and the structure has been solved by the molecular-replacement method using a previously built theoretical model of the protein as the search molecule. PMID- 14747730 TI - Crystallization and preliminary X-ray crystallographic studies on recombinant rat choline acetyltransferase. AB - Choline acetyltransferase (ChAT) catalyzes the biosynthesis of the neurotransmitter acetylcholine from acetyl-CoA and choline in cholinergic neurons. Rat ChAT (rChAT) was overexpressed in Escherichia coli, purified by affinity chromatography and crystallized. Diffraction data were collected from a single crystal under cryoconditions at the F1 beamline at the Cornell High Energy Synchrotron Source, with a maximal useful diffraction pattern to 1.55 A resolution. The crystals were shown to belong to the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 138.97, b = 77.67, c = 59.67 A and a scaling R(sym) of 0.054 for 72 446 unique reflections. Packing considerations indicate there to be one molecule per asymmetric unit. It is expected that in the near future the structure of rChAT will be obtained using molecular-replacement methods. Elucidation of the structure of rChAT will aid in the development of therapeutic agents for Alzheimer's disease. PMID- 14747731 TI - Crystals of the ribonucleotide reductase R2 protein from Chlamydia trachomatis obtained by heavy-atom co-crystallization. AB - Ribonucleotide reductases (RNRs) catalyse the conversion of ribonucleotides to deoxyribonucleotides, utilizing radical chemistry to carry out the reaction. Class I RNRs consist of R1 and R2 subunits: R1 contains the active site and R2 generates and stores a stable tyrosyl radical. The conserved tyrosine where the radical is stored until needed in R1 has previously been believed to be an absolute requirement for R2 activity. The Chlamydia trachomatis R2 lacks this tyrosine and a phenylalanine is present in its place, but the protein is still active. Here, the crystallization of C. trachomatis R2 is described. A heavy-atom co-crystallization approach was used to obtain crystals. Hopefully, the C. trachomatis R2 structure will provide key clues as to how this enzyme is able to function while lacking the features that have previously been believed to be essential for activity. PMID- 14747732 TI - Crystallization and preliminary X-ray crystallographic analysis of the RecR protein from Deinococcus radiodurans, a member of the RecFOR DNA-repair pathway. AB - The RecR protein plays a key role in the RecFOR pathway of recombination, which is necessary for the repair of ssDNA gaps. RecR from Deinococcus radiodurans has been overexpressed in Escherichia coli and crystallized at 297 K using polyethylene glycol 1000 as a precipitant. X-ray diffraction data to 2.90 A resolution have been collected at 100 K using Cu Kalpha X-rays from a mercury soaked crystal. The crystal belongs to space group C222(1), with unit-cell parameters a = 106.96, b = 122.25, c = 156.01 A. The asymmetric unit contains four monomers of RecR, with a crystal volume per protein weight (V(M)) of 2.57 A(3) Da(-1) and a solvent content of 51.0%. PMID- 14747733 TI - Crystallization and preliminary X-ray crystallographic analyses of CMY-1 and CMY 10, plasmidic class C beta-lactamases with extended substrate spectrum. AB - Plasmid-encoded class C beta-lactamases, including CMY-1 and CMY-10, hydrolyze the lactam bonds of beta-lactam antibiotics, inducing therapeutic failure and a lack of eradication of clinical isolates by third-generation cephalosporins or cephamycins. Therefore, the enzymes are potential targets for developing agents against pathogens isolated from patients suffering from wound infection, urinary tract infection or pneumonia. CMY-1 and CMY-10 were purified and crystallized at 298 K. X-ray diffraction data from CMY-1 and CMY-10 crystals have been collected to 2.5 and 1.5 A resolution, respectively, using synchrotron radiation. The crystals of the two proteins are isomorphous and belong to the primitive monoclinic space group P2(1). PMID- 14747734 TI - Crystallization and preliminary X-ray analysis of an acetone carboxylase from Xanthobacter autotrophicus strain Py2. AB - Acetone carboxylase from Xanthobacter autotrophicus strain Py2 catalyzes the MgATP-dependent carboxylation of acetone to acetoacetate. Interestingly, during this reaction ATP is hydrolyzed to AMP and inorganic phosphate, suggesting a novel carboxylation mechanism. Acetone carboxylase is a heterohexameric protein comprised of three different polypeptides having molecular weights of 86 342, 78 509 and 19 773 Da arranged in an alpha(2)beta(2)gamma(2) quaternary structure. Here, the crystallization and preliminary X-ray data analysis of acetone carboxylase is reported. The acetone carboxylase isolated from the aerobic microorganism X. autotrophicus strain Py2 crystallizes in a primitive orthorhombic point group P222, with unit-cell parameters a = 76.2, b = 122.0, c = 264.2 A. The Matthews coefficient calculation indicates that one alphabetagamma half of the large protein complex is located in the asymmetric unit in this crystal form. Crystals have been obtained that diffract to better than 2.8 A resolution and data have been collected to 3.2 A resolution. PMID- 14747735 TI - Crystallization of 4-hydroxybenzoyl-CoA reductase and the structure of its electron donor ferredoxin. AB - 4-Hydroxybenzoyl-CoA reductase (4-HBCR) is a central enzyme in the metabolism of phenolic compounds in anaerobic bacteria. The enzyme catalyzes the reductive removal of the phenolic hydroxyl group from 4-hydroxybenzoyl-CoA, yielding benzoyl-CoA and water. 4-HBCR belongs to the xanthine oxidase (XO) family of molybdenum enzymes which occur as heterodimers, (alphabetagamma)(2). 4-HBCR contains two molybdopterins, four [2Fe-2S] and two [4Fe-4S] clusters and two FADs. A low-potential Allochromatium vinosum-type ferredoxin containing two [4Fe 4S] clusters serves as an in vivo electron donor for 4-HBCR. In this work, the oxygen-sensitive proteins 4-HBCR and the ferredoxin (TaFd) from Thauera aromatica were crystallized under anaerobic conditions. 4-HBCR crystallized with PEG 4000 and MPD as precipitant diffracted to about 1.6 A resolution and the crystals were highly suitable for X-ray structure analysis. Crystals of TaFd were obtained with (NH(4))(3)PO(4) as precipitant and revealed a solvent content of 77%, which is remarkably high for a small soluble protein. The structure of TaFd was solved at 2.9 A resolution by the molecular-replacement method using the highly related structure of the ferredoxin (CvFd) from A. vinosum as a model. Structural changes between the two ferredoxins around the [4Fe-4S] cluster can be correlated with their different redox potentials. PMID- 14747736 TI - Structural analysis of neprilysin with various specific and potent inhibitors. AB - Neutral endopeptidase (NEP) is the major enzyme involved in the metabolic inactivation of a number of bioactive peptides including the enkephalins, substance P, endothelin, bradykinin and atrial natriuretic factor. Owing to the physiological importance of NEP in the modulation of nociceptive and pressor responses, there is considerable interest in inhibitors of this enzyme as novel analgesics and antihypertensive agents. Here, the crystal structures of the soluble extracellular domain of human NEP (residues 52-749) complexed with various potent and competitive inhibitors are described. The structures unambiguously reveal the binding mode of the different zinc-chelating groups and the subsite specificity of the enzyme. PMID- 14747737 TI - Refinement of Haemophilus influenzae diaminopimelic acid epimerase (DapF) at 1.75 A resolution suggests a mechanism for stereocontrol during catalysis. AB - Diaminopimelate (DAP) epimerase (DapF) is central to the biosynthesis of both lysine and cell-wall peptidoglycan in many bacteria species. The peptidoglycan layer provides great potential for the development of novel antimicrobials as it is a uniquely prokaryotic feature. Crystals of recombinant Haemophilus influenzae DapF that diffract to beyond 2 A resolution have been obtained which facilitated the solution of the structure by molecular replacement at a resolution approximately 1 A higher than that previously determined. An analysis of the structure (i) in comparison to other PLP-independent racemaces and (ii) in relation to the catalytic mechanism and stereospecificity of DapF is presented. PMID- 14747738 TI - Structure of the saccharide-binding domain of the human natural killer cell inhibitory receptor p75/AIRM1. AB - The high-resolution crystal structure of the functional N-terminal domain from the extracellular region of the human natural killer cell inhibitory receptor p75/AIRM1 or Siglec-7 has been determined at 1.45 A resolution; it was obtained from a crystal belonging to a primitive monoclinic space group, with unit-cell parameters a = 32.65, b = 49.72, c = 39.79 A, alpha = gamma = 90, beta = 113 degrees. The structure reported here belongs to a different space group than the previously described Siglec-7 structure and was obtained using a bacterial expression system. The structure unveils the fine structural requirements adopted by a natural killer cell inhibitory receptor of the Siglec family in target-cell recognition and binding. PMID- 14747740 TI - Pathophysiology of cerebral ischemia and brain trauma: similarities and differences. AB - Current knowledge regarding the pathophysiology of cerebral ischemia and brain trauma indicates that similar mechanisms contribute to loss of cellular integrity and tissue destruction. Mechanisms of cell damage include excitotoxicity, oxidative stress, free radical production, apoptosis and inflammation. Genetic and gender factors have also been shown to be important mediators of pathomechanisms present in both injury settings. However, the fact that these injuries arise from different types of primary insults leads to diverse cellular vulnerability patterns as well as a spectrum of injury processes. Blunt head trauma produces shear forces that result in primary membrane damage to neuronal cell bodies, white matter structures and vascular beds as well as secondary injury mechanisms. Severe cerebral ischemic insults lead to metabolic stress, ionic perturbations, and a complex cascade of biochemical and molecular events ultimately causing neuronal death. Similarities in the pathogenesis of these cerebral injuries may indicate that therapeutic strategies protective following ischemia may also be beneficial after trauma. This review summarizes and contrasts injury mechanisms after ischemia and trauma and discusses neuroprotective strategies that target both types of injuries. PMID- 14747741 TI - A model of global cerebral ischemia in C57 BL/6 mice. AB - A reproducible model of global cerebral ischemia in mice is essential for elucidating the molecular mechanism of ischemic neuronal injury. Such a model is particularly important in the mouse because many genetically engineered mutant animals are available. In C57BL/6 and SV129/EMS mice, we evaluated a three-vessel occlusion model. Occlusion of the basilar artery with a miniature clip was followed by bilateral carotid occlusion. The mean cortical cerebral blood flow was reduced to less than 10% of the preischemic value, and the mean anoxic depolarization was attained within 1 minute. In C57BL/6 mice, there was CA1 hippocampal neuronal degeneration 4 days after ischemia. Neuronal damage depended upon ischemic duration: the surviving neuronal count was 78.5 +/- 8.5% after 8 minute ischemia and 8.4 +/- 12.7% after 14-minute ischemia. In SV129/EMS mice, similar neuronal degeneration was not observed after 14-minute ischemia. The global ischemia model in C57BL/6 mice showed high reproducibility and consistent neuronal injury in the CA1 sector, indicating that comparison of ischemic outcome between wild-type and mutant mice could provide meaningful data using the C57BL/6 genetic background. Strain differences in this study highlight the need for consideration of genetic background when evaluating ischemia experiments in mice. PMID- 14747742 TI - Intracerebral hirudin injection attenuates ischemic damage and neurologic deficits without altering local cerebral blood flow. AB - There has been considerable interest in the use of thrombin inhibitors to reduce the occurrence of stroke or to potentiate tissue plasminogen activator-induced reperfusion. However, there is growing evidence that thrombin may also have extravascular effects that influence ischemic brain injury. Male Sprague-Dawley rats were subjected to either 90 minutes of temporary middle cerebral artery (MCA) occlusion or sham operation to examine thrombin and protease activated receptor-1 (PAR-1) expression. In another set of rats, the MCA was occluded for 90 minutes and 10 U of hirudin or the same volume of vehicle was injected into the caudate followed by reperfusion for up to 28 days, to test the effects of local thrombin inhibition on ischemic damage, neurologic outcome and cerebral blood flow (CBF). Thrombin immunoreactivity was increased in the ischemic caudate at 4 and 24 hours, whereas PAR-1 expression was unchanged. Hirudin reduced infarct volume in the caudate at 24 hours (79 +/- 41 vs. 115 +/- 20 mm3, P < 0.05) and resulted in a larger residual tissue volume in the caudate at 28 days (17.6 +/- 3.9 vs. 11.8 +/- 6.3 mm3, P < 0.05). Hirudin treatment also had a beneficial effect on body weight and ameliorated neurologic deficits tested by forelimb placing and forelimb use asymmetry during 28 days survival. These beneficial effects of hirudin were not associated with improved regional CBF during reperfusion. These results suggest that, in addition to their effects on coagulation and circulation, thrombin inhibitors also have direct neuroprotective properties and may be considered in stroke therapy. PMID- 14747743 TI - Time course and cellular distribution of hsp27 and hsp72 stress protein expression in a quantitative gerbil model of ischemic injury and tolerance: thresholds for hsp72 induction and hilar lesioning in the context of ischemic preconditioning. AB - The distribution and time course of expression of the heat shock/stress proteins, hsp27 and hsp72, were evaluated in a highly controlled gerbil model of ischemic injury and tolerance induction, in which the duration of ischemic depolarization in each hippocampus provides a precise quantitative index of insult severity. Gerbils were subjected to brief priming insults (2- to 3.5-minute depolarization) that produce optimal preconditioning, to severe test insults (6- to 8.5-minute depolarization) that produce complete CA1 neuron loss in naive animals, or to combined insults administered 1 week apart, after which almost complete tolerance to CA1 neuron injury is observed. Immunoreactivities of hsp27, hsp72, glial fibrillary acidic protein and microtubule-associated protein 2 (MAP2) were evaluated in animals perfused at defined intervals after the final insult in each treatment group, using a variation of established antigen-retrieval procedures that significantly improves detection of many proteins in vibratome brain sections. Hsp72 was detected in CA1 neurons of some hippocampi 2 to 4 days after preconditioning, but this was only seen after the longest priming depolarizations, whereas shorter insults that still induced optimal tolerance failed to induce hsp72. Hsp72 was induced after test insults in preconditioned hippocampi, but at a higher depolarization threshold than observed for naive animals. An astrocytic localization of hsp27 was observed in regions of neuron injury, as indicated by reduced MAP2 immunoreactivity, and was primarily restricted to dentate hilus after preconditioning insults. These results establish that limited hilar lesions are characteristic of optimal preconditioning, whereas prior neuronal expression of either hsp72 or hsp27 is not required for ischemic tolerance. PMID- 14747744 TI - Task repetition can affect functional magnetic resonance imaging-based measures of language lateralization and lead to pseudoincreases in bilaterality. AB - Repeated functional magnetic resonance imaging (fMRI) during learning and recovery can inform us about functional reorganization in the brain. We examined how, in the absence of reorganization, simple task repetition affects measures of fMRI activation. We studied fMRI activation over 10 consecutive sessions of silent word generation in a healthy subject. Additionally, we performed functional transcranial Doppler sonography (fTCD) to learn about the temporal pattern of corresponding changes in cerebral blood flow velocity (CBFV) and pulsatility. With repetition, word generation-associated increases in heart rate diminished steadily. Task repetition also led to a net increase in CBFV bilaterally and to a bilateral increase in the number of activated voxels on fMRI. As a result, whereas the absolute interhemispheric difference of activated voxels remained constant, there was a decrease in the standard fMRI index for language lateralization [LIfMRI = 100 (activated voxels in left hemisphere - voxels in right)/(voxels in left + voxels in right)]. Thus task repetition can lead to changes in task-related autonomic drive and an augmentation of bihemispheric blood flow. This can mimic increasing bilaterality of brain activation. PMID- 14747745 TI - Effect of astrocytic energy metabolism depressant on 14C-acetate uptake in intact rat brain. AB - Fluorocitrate, a selective astrocytic toxin, was microinjected into the right striatum of rat brain, and the regional distribution of 14C-acetate was measured using autoradiography. A significant reduction (more than 80%) in 14C-acetate uptake over a 5-minute period was observed in the right striatum, compared with that in the left striatum (saline infused), 4 hours after fluorocitrate (1 nmol/microL) infusion. This effect was transient, and 14C-acetate uptake had almost returned to normal at 24 hours after the fluorocitrate infusion. In contrast, the regional blood flow in the striatum, as determined using 14C iodoamphetamine, was significantly increased by the fluorocitrate infusion. The present observations indicate that 14C-acetate uptake might be a useful characteristic for examining astrocytic energy metabolism in the intact brain. PMID- 14747746 TI - Defining ischemic burden after traumatic brain injury using 15O PET imaging of cerebral physiology. AB - Whereas postmortem ischemic damage is common in head injury, antemortem demonstration of ischemia has proven to be elusive. Although 15O positron emission tomography may be useful in this area, the technique has traditionally analyzed data within regions of interest (ROIs) to improve statistical accuracy. In head injury, such techniques are limited because of the lack of a priori knowledge regarding the location of ischemia, coexistence of hyperaemia, and difficulty in defining ischemic cerebral blood flow (CBF) and cerebral oxygen metabolism (CMRO2) levels. We report a novel method for defining disease pathophysiology following head injury. Voxel-based approaches are used to define the distribution of oxygen extraction fraction (OEF) across the entire brain; the standard deviation of this distribution provides a measure of the variability of OEF. These data are also used to integrate voxels above a threshold OEF value to produce an ROI based upon coherent physiology rather than spatial contiguity (the ischemic brain volume; IBV). However, such approaches may suffer from poor statistical accuracy, particularly in regions with low blood flow. The magnitude of these errors has been assessed in modeling experiments using the Hoffman brain phantom and modified control datasets. We conclude that this technique is a valid and useful tool for quantifying ischemic burden after traumatic brain injury. PMID- 14747747 TI - Incidence and mechanisms of cerebral ischemia in early clinical head injury. AB - Antemortem demonstration of ischemia has proved elusive in head injury because regional CBF reductions may represent hypoperfusion appropriately coupled to hypometabolism. Fifteen patients underwent positron emission tomography within 24 hours of head injury to map cerebral blood flow (CBF), cerebral oxygen metabolism (CMRO2), and oxygen extraction fraction (OEF). We estimated the volume of ischemic brain (IBV) and used the standard deviation of the OEF distribution to estimate the efficiency of coupling between CBF and CMRO2. The IBV in patients was significantly higher than controls (67 +/- 69 vs. 2 +/- 3 mL; P < 0.01). The coexistence of relative ischemia and hyperemia in some patients implies mismatching of perfusion to oxygen use. Whereas the saturation of jugular bulb blood (SjO2) correlated with the IBV (r = 0.8, P < 0.01), SjO2 values of 50% were only achieved at an IBV of 170 +/- 63 mL (mean +/- 95% CI), which equates to 13 +/- 5% of the brain. Increases in IBV correlated with a poor Glasgow Outcome Score 6 months after injury (rho = -0.6, P < 0.05). These results suggest significant ischemia within the first day after head injury. The ischemic burden represented by this "traumatic penumbra" is poorly detected by bedside clinical monitors and has significant associations with outcome. PMID- 14747748 TI - Genome-wide gene expression analysis for induced ischemic tolerance and delayed neuronal death following transient global ischemia in rats. AB - Genome-wide gene expression analysis of the hippocampal CA1 region was conducted in a rat global ischemia model for delayed neuronal death and induced ischemic tolerance using an oligonucleotide-based DNA microarray containing 8,799 probes. The results showed that expression levels of 246 transcripts were increased and 213 were decreased following ischemia, corresponding to 5.1% of the represented probe sets. These changes were divided into seven expression clusters using hierarchical cluster analysis, each with distinct conditions and time-specific patterns. Ischemic tolerance was associated with transient up-regulation of transcription factors (c-Fos, JunB Egr-1, -2, -4, NGFI-B), Hsp70 and MAP kinase cascade-related genes (MKP-1), which are implicated cell survival. Delayed neuronal death exhibited complex long-lasting changes of expression, such as up regulation of proapoptotic genes (GADD153, Smad2, Dral, Caspase-2 and -3) and down-regulation of genes implicated in survival signaling (MKK2, and PI4 kinase, DAG/PKC signaling pathways), suggesting an imbalance between death and survival signals. Our study provides a differential gene expression profile between delayed neuronal death and induced ischemic tolerance in a genome-wide analysis, and contributes to further understanding of the complex molecular pathophysiology in cerebral ischemia. PMID- 14747749 TI - Restriction-mediated differential display (RMDD) identifies pip92 as a pro apoptotic gene product induced during focal cerebral ischemia. AB - Studies of gene expression changes after cerebral ischemia can provide novel insight into ischemic pathophysiology. Here we describe application of restriction-mediated differential display to screening for differentially expressed genes after focal cerebral ischemia. This method combines the nonredundant generation of biotin-labeled fragment sets with the excellent resolution of direct blotting electrophoresis, reliable fragment recovery, and a novel clone selection strategy. Using the filament model in mouse with 90 minutes MCA occlusion followed by 2, 6, and 20 hours reperfusion, we have compared gene expression in sham-operated animals to both the ipsi- and contralateral forebrain hemisphere of ischemic mice. Our screening method has resulted in the identification of 70 genes differentially regulated after transient middle cerebral artery occlusion (MCAO), several of which represent unknown clones. We have identified many of the previously published regulated genes, lending high credibility to our method. Surprisingly, we detected a high degree of correspondent regulation of genes in the nonischemic hemisphere. A high percentage of genes coding for proteins in the respiratory chain was found to be up-regulated after ischemia, potentially representing a new mechanism involved in counteracting energy failure or radical generation in cerebral ischemia. One particularly interesting gene, whose upregulation by ischemia has not been described before, is pip92; this gene shows a rapid and long-lasting induction after cerebral ischemia. Here we demonstrate that pip92 induces cell death in primary neurons and displays several hallmarks of pro-apoptotic activity upon overexpression, supporting the notion that we have identified a novel pathophysiological player in cerebral ischemia. In summary, restriction-mediated differential display has proven its suitability for screening complex samples such as brain to reliably identify regulated genes, which can uncover novel pathophysiological mechanisms. PMID- 14747750 TI - Vascular endothelial growth factor induces abnormal microvasculature in the endoglin heterozygous mouse brain. AB - Hereditary hemorrhagic telangiectasia (HHT), associated with brain arteriovenous malformations, is caused by a loss of function mutation in either the endoglin (HHT1) or activin receptor-like kinase 1 gene (ALK-1, HHT2). Endoglin heterozygous (Eng+/-)mice have been proposed as a disease model. To better understand the role of endoglin in vascular malformation development, we examined the effect of vascular endothelial growth factor (VEGF) hyperstimulation on microvessels in adult endoglin heterozygous (Eng+/-) mice using an adenoviral vector to deliver recombinant human VEGF165 cDNA (AdhVEGF) into basal ganglia. VEGF expression was increased in AdhVEGF mice compared with the AdlacZ and saline group (P < 0.05) and localized to multiple cell types (neurons, astrocytes, endothelial cells, and smooth muscle cells) by double-labeled immunostaining. VEGF overexpression increased microvessel count for up to 4 weeks in both the Eng+/+ and Eng+/- groups (Eng+/+ 185 +/- 14 vs. Eng+/- 201 +/- 10 microvessels/mm2). Confocal microscopic examination revealed grossly abnormal microvessels in eight of nine Eng+/- mouse brains compared with zero of nine in Eng+/+ mice (P < 0.05). Abnormal microvessels featured enlargement, clustering, twist, or spirals. VEGF receptor Flk-1 and TGF-beta receptor 1 (T beta R1) expression were reduced in the Eng+/- mouse brain compared with control. Excessive VEGF stimulation may play a pivotal role in the initiation and development of brain vessel malformations in states of relative endoglin insufficiency in adulthood. These observations are relevant to our general understanding of the maintenance of vascular integrity. PMID- 14747751 TI - Identification of potential stroke targets by lentiviral vector mediated overexpression of HIF-1 alpha and HIF-2 alpha in a primary neuronal model of hypoxia. AB - The identification of genes differentially regulated by ischemia will lead to an improved understanding of cell death pathways such as those involved in the neuronal loss observed following a stroke. Furthermore, the characterization of such pathways could facilitate the identification of novel targets for stroke therapy. We have used a novel approach to amplify differential gene expression patterns in a primary neuronal model of stroke by employing a lentiviral vector system to specifically bias the transcriptional activation of hypoxically regulated genes. Overexpression of the hypoxia-induced transcription factor subunits HIF-1 alpha and HIF-2 alpha elevated hypoxia-mediated transcription of many known HIF-regulated genes well above control levels. Furthermore, many potentially novel HIF-regulated genes were discovered that were not previously identified as hypoxically regulated. Most of the novel genes identified were activated by a combination of HIF-2 alpha overexpression and hypoxic insult. These included several genes with particular importance in cell survival pathways and of potential therapeutic value. Hypoxic induction of HIF-2 alpha may therefore be a critical factor in mediating protective responses against ischemic injury. Further investigation of the genes identified in this study may provide increased understanding of the neuronal response to hypoxia and may uncover novel therapeutic targets for the treatment of cerebral ischemia. PMID- 14747752 TI - Mechanisms of erythropoietin-induced brain protection in neonatal hypoxia ischemia rat model. AB - Erythropoietin, a hemotopoietic growth factor, has brain protective actions. This study investigated the mechanisms of Recombinant Human EPO (rhEPO)-induced brain protection in neonates. An established rat hypoxia-ischemia model was used by ligation of the right common carotid artery of 7-day-old pups, followed by 90 minute of hypoxia (8% 02 and 92% N2) at 37 degrees C. Animals were divided into three groups: control, hypoxia-ischemia, and hypoxia-ischemia plus rhEPO treatment. In rhEPO treated pups, 300 units rhEPO was administered intraperitoneally 24 hours before hypoxia. rhEPO treatment (300 units) was administered daily for an additional 2 days. ELISA and immunohistochemistry examined the expression of EPO and EPOR. Brain weight, morphology, TUNEL assay, and DNA laddering evaluated brain protection. rhEPO abolished mortality (from 19% to 0%) during hypoxia insult, increased brain weight from 52% to 88%, reduced DNA fragmentation, and decreased TUNEL-positive cells. Real-time RT-PCR, Western blot, and immunohistochemistry revealed an enhanced expression of heat shock protein 27 (HSP27) in ischemic brain hemisphere. Double labeling of TUNEL with HSP27 showed most HSP27 positive cells were negative to TUNEL staining. rhEPO reduces brain injury, especially apoptotic cell death after neonatal hypoxia ischemia, partially mediated by the activation of HSP27. PMID- 14747753 TI - Myopia: pathways to therapy. PMID- 14747755 TI - Community-based study of the association of high myopia in children with ocular and systemic disease. AB - PURPOSE: High myopia in childhood is associated with important ocular and systemic conditions. However in the UK, high myopia in early childhood is not specifically identified in current ophthalmology, optometry, or orthoptic protocols for screening, referral, or investigation. An ongoing study in the West Midlands, UK, is investigating high myopia presenting to community health care clinics with the aim of compiling guidelines for assessment and subsequent referral. METHODS: Children with high myopia were identified from community optometric and orthoptic sources and invited for an ophthalmology and optometry examination to ascertain possible ocular or systemic disease. RESULTS: High myopia with no associated ocular or systemic condition was present in 15 (56%) of the children. In seven children (25%), associated ocular problems were found including unrecognized retinal dystrophies and amblyopia. Systemic disorders associated with high myopia were found in five children (19%) and included Sticklers syndrome, Weill-Marchesani syndrome, and homocystinuria. In one child, the diagnosis made before this study was found to be incorrect, and in another child, the results were inconclusive. In two cases, the diagnosis of a systemic condition in the child led to the identification of the disease in at least one relative. CONCLUSIONS: There is a high prevalence of ocular and systemic abnormality in young children seen in the community. Optometric and ophthalmologic assessment of children less than 10 years with myopia > or =5 D is likely to identify significant ocular or systemic disease, a proportion of which will respond to medical intervention. Detection and prompt referral of these cases by community health care services may be expected to prolong vision and possibly life expectancy. PMID- 14747754 TI - Prevalence and distribution of corrective lenses among school-age children. AB - BACKGROUND: No population-based data are available regarding the proportion of school-age children who have corrective lenses in the U.S. The objective of this study was to quantify the proportion of children who have corrective lenses (glasses or contact lenses) and to evaluate the association of corrective lenses with age, gender, race/ethnicity, health insurance status, and family income. METHODS: Children 6 to 18 years of age were identified in the 1998 Medical Expenditure Panel Survey. National estimates were made of the proportion with corrective lenses. Logistic regression modeling was used to assess factors that were associated with corrective lenses. RESULTS: Based on the 5,141 children in the 1988 Medical Expenditure Panel Survey, an estimated 25.4% of the 52.6 million children between 6 and 18 years had corrective lenses. Girls had greater odds than boys of having corrective lenses (odds ratio, 1.41; p < 0.001). Insured children, regardless of race/ethnicity, and uninsured nonblack/non-Hispanic children had similar odds of having corrective lenses. Compared with uninsured black or Hispanic children (odds ratio, 1), greater odds of corrective lens use was found among uninsured nonblack/non-Hispanic children (odds ratio, 2.29; p = 0.002) and black or Hispanic children with public (odds ratio, 1.67; p = 0.005) or private health insurance (odds ratio,1.77; p = 0.004). Among families with an income > or =200% of the federal poverty level, the odds of having corrective lenses increased with age (p < or = 0.04). In contrast, among those families <200% of the federal poverty level, the odds of having corrective lenses at 12 to 14 years was similar to 15- to 18-year olds (p = 0.93). CONCLUSIONS: The use of corrective lenses suggests that correctable visual impairment is the most common treatable chronic condition of childhood. Income, gender, and race/ethnicity, depending on insurance status, are associated with having corrective lenses. The underlying causes and the impacts of these differences must be understood to ensure optimal delivery of eye care. PMID- 14747756 TI - Unequal reduction in visual acuity with positive and negative defocusing lenses in myopes. AB - Myopes have a reduced accommodative response to negative lenses compared with nonmyopes. Mathematical models predict that the reduced accommodative response is due to a decrease in sensitivity to blur in myopes. We examined the effect of blur induced by positive and negative defocusing lenses on visual acuity in 12 myopes and 12 nonmyopes during cycloplegia for up to +/-3.00 D of defocus in 0.25 D steps. Although nonmyopes showed a symmetrical reduction in visual acuity with positive and negative lenses, the myopic group showed less acuity loss with negative lenses compared with positive lenses. The magnitude of visual acuity loss was lower with negative lenses in myopes compared with nonmyopes. No significant difference in visual acuity with positive lenses was found between myopes and nonmyopes. Residual accommodation (after cycloplegia) was about 0.20 D in both myopes and nonmyopes and was too small to explain the relatively good visual acuity through minus lenses in the myopic group. The reduced accommodative response known to occur in myopes may be due to the relatively small effect that negative lens blur has on their visual acuity. PMID- 14747757 TI - Astigmatic axis is related to the level of spherical ametropia. AB - PURPOSE: Against-the-rule (ATR) astigmatism has been shown to be a risk factor for subsequent myopia development. In this study, we evaluated the relationship between astigmatic axis and the level of spherical ametropia in both myopes and hypermetropes. METHODS: Astigmatic axes were analyzed in two distinct cohorts. First, 53 high myopes from families that were recruited for linkage analysis were compared with an age-matched control group derived from family members. Second, cross-sectional data were analyzed for 90,884 subjects attending 19 optometric practices in the north of England. Initially, the relationship between astigmatic axis and cylinder power and between axis and sphere power were analyzed in 21- to 40-year olds and 21- to 30-year olds, respectively, to control for the effects of age. Multivariate logistic regression analysis was then performed using data for all compound astigmats in the cohort to examine the effect of sphere power, cylinder power, age, and sex on the odds of subjects having either ATR or with the-rule (WTR) astigmatism. RESULTS: In the genetic study cohort, there was an excess of WTR astigmats in the high myopes compared with controls, but this only reached significance for the right eye. In the much larger optometric practice sample, the association of WTR astigmatism with high myopia was highly significant. A parallel increase in WTR astigmatism was also found for high hypermetropes. In addition, the odds of having WTR astigmatism were increased if subjects were young or had a high cylinder power. ATR astigmatism occurred more often with increasing age and in subjects with lower spherical ametropia. Indeed, for 21- to 30-year-old subjects with low myopia (> or =-2.00 DS in the least minus meridian), ATR occurred more often than WTR astigmatism. CONCLUSION: Astigmatic axis was found to be related to the level of ametropia, with both a higher spherical component or higher cylinder power increasing the odds of astigmatism being WTR. Low ametropes, particularly myopes, were more likely to have axes ATR. PMID- 14747758 TI - Ocular component measurement using the Zeiss IOLMaster. AB - PURPOSE: Axial length is traditionally measured using A-scan ultrasound. The IOLMaster is a new instrument that uses partial coherence interferometry to measure axial length. We compared the repeatability of these techniques for both an experienced and an inexperienced observer, the agreement between the two techniques, and the effect of cycloplegia on IOLMaster measurements. METHODS: Five measurements of axial length and three measurements of anterior chamber depth were taken with the IOLMaster in two sessions separated by 1 to 12 days in 20 young adults. The two examiners each took measurements, and the subject was then cyclopleged with 1% tropicamide. The IOLMaster readings were then repeated by both examiners, followed by five ultrasound readings. Repeatability was evaluated by calculating the difference between measurements from the two sessions. The mean and standard deviation of these differences was then used to determine the 95% limits of agreement (LoA) for each technique. In addition, the agreement between the IOLMaster and ultrasound was assessed, along with the effect of cycloplegia on IOLMaster readings. RESULTS: The IOLMaster was more repeatable than ultrasound. For axial length, the 95% LoA were -0.11 to +0.07 mm, -0.06 to +0.05 mm, and -0.25 to +0.35 mm, for noncycloplegic IOLMaster, cycloplegic IOLMaster, and ultrasound, respectively. The two instruments showed modest agreement with each other (mean difference, +0.12 mm; 95% LoA, -0.39 to +0.64 mm; p > 0.0125). Cycloplegia had no significant effect on IOLMaster axial length measurements. The 95% LoA for anterior chamber depth measurement were 0.11 to +0.18 mm, -0.06 to +0.04 mm, and -0.19 to +0.21 mm, for noncycloplegic IOLMaster, cycloplegic IOLMaster, and ultrasound, respectively. The IOLMaster gave significantly longer anterior chamber depths than ultrasound (mean, +0.18 mm; 95% LoA, -0.02 to +0.37 mm; p < 0.0125), and cycloplegia produced significantly deeper anterior chamber depths using the IOLMaster (mean, +0.12 +/- 0.09 mm; 95% LoA, -0.05 to +0.29 mm; t = 6.17; p < 0.001). The experienced observer's measurements were more repeatable than the inexperienced observer's for ultrasound, but not for the IOLMaster. CONCLUSIONS: The superior repeatability of the IOLMaster suggests that it should become the standard for axial length measurement. The 95% limits of agreement for the cycloplegic measurements correspond to a change in refractive error of +/-0.12 D. PMID- 14747759 TI - Variability of the ocular component measurements in children using A-scan ultrasonography. AB - Knowledge of the precision with which the various ocular components can be measured with available techniques is vital to our ability to track changes in the anatomy of the eye in relation to the development of refractive error. Corneal touch A-scan ultrasound measurements were obtained by either an instrument-mounted or by a hand-held method on 469 children aged 6 to 11 years at their enrollment into the Correction of Myopia Evaluation Trial. Variability of measurements was calculated for overall axial length, anterior chamber depth, lens thickness, and vitreous chamber depth. The mean variability of overall axial length was 0.062 +/- 0.043 mm and 0.061 +/- 0.056 mm for the right and left eyes, respectively. Mean variability of the ocular component measures was similar. The mean variability was not influenced by the age, gender, level of myopia, or residual accommodation of the subjects. Statistically significant differences between the instrument-mounted and the hand-held method and among certain ethnic groups were found, but the differences were not of a magnitude to be clinically significant. A-scan ultrasonography is sensitive to changes in the axial length and vitreous chamber depth equivalent to 0.25 D and is therefore a useful technique to assess changes in these ocular components in children. The precision of lens thickness is poorer than the equivalent of 1.0 D, and, therefore, A-scan may not be sufficiently precise to be useful in studies of active accommodation or lens growth. PMID- 14747760 TI - The Nepal longitudinal study: predicting myopia from the rate of increase in vitreous chamber depth. AB - Biometric data on 897 Tibetan children living in Kathmandu, Nepal were collected over the period 1992 to 2000 with regular visits every 2 years. Measurements included cycloplegic autorefraction, A-scan ultrasonography, and video phakometry. Children who had not been studied at least once at age 12 years or older were not included in these analyses. The other subjects were divided into two groups; myopic if the refractive error was ever <-0.50 D, and emmetropic/hyperopic if the refractive error was never <-0.50 D, the nonmyopic group. Using all children who had been examined with four or five observations over time, the change of vitreous chamber depth with age by group was determined using a mixed-model regression method. The increase in vitreous length was 0.070 mm/year for the emmetropic group and 0.165 mm/year for the myopic group, with the differences apparent before the onset of myopia. An independent group of 59 children in whom there were two vitreous chamber depth measures before the age of 12 years and one measure taken after 12 years of age were used to assess the rate of increase in vitreous chamber depth as a predictor of myopia. Two other methods were examined using the independent group; the ratio of axial length to corneal radius of curvature and refractive error at age 10 years. Predictors based on rate of increase in vitreous chamber depth and axial length/corneal radius of curvature had sensitivities of 75% and 45%, respectively, and refractive error at age 10 years as a predictor for those who will not become myopic had a sensitivity of 88%. PMID- 14747761 TI - Accuracy of noncycloplegic autorefraction in school-age children in China. AB - PURPOSE: To evaluate the accuracy of noncycloplegic autorefraction in a representative sample of school-age children in China. METHODS: Refractive error was measured with an autorefractor, both before and after cycloplegia induced with cyclopentolate, in a population-based sample of 4973 children between the ages of 7 and 18 years. Spherical equivalent refractive error and astigmatism as represented by Jackson crossed-cylinders (J0 and J45) were the main outcome measures. RESULTS: Noncycloplegic measurements of equivalent spheres were consistently more negative or less positive than those after cycloplegia, with mean +/- SD differences of -1.23 +/- 0.97 D. The differences were particularly large for hyperopic eyes (mean difference of -2.98 +/- 1.65 D for hyperopia of at least +2.00 D) while becoming progressively smaller for emmetropic eyes, and smaller yet for myopic eyes (mean difference of -0.41 +/- 0.46 D for myopia of 2.00 D or more). Increasing age was associated with increased, but clinically insignificant, differences. Little difference was found between noncycloplegic and cycloplegic measurements of astigmatism: mean J0 and J45 differences were 0.08 +/- 0.13 D and -0.01 +/- 0.09 D, respectively. CONCLUSIONS: Noncycloplegic autorefraction was found to be highly inaccurate in school-age children and, thus, not suitable for studies of refractive error or for prescription of glasses in this population. PMID- 14747762 TI - Comparison of spherical equivalent refraction and astigmatism measured with three different models of autorefractors. AB - PURPOSE: The purpose of this study was to compare refractions measured with three different autorefractors. METHODS: The refractive error of each eye of 50 adults aged 17 to 59 years (mean, 30.5 years) was measured without cycloplegia using the Canon R-1 and two newer instruments, the Grand Seiko WR-5100K and the Nidek ARK 700-A. For the first two, an isolated line of 20/100 letters on an ETDRS chart at 4.0 m served as a target, whereas for the Nidek, the subject looked at a picture of a balloon in the instrument. Five readings were taken for each eye, and the data (sphere, negative cylinder power, and axis) were analyzed using Fourier decomposition of the power profile. Each reading was broken down into the spherical equivalent (M) and two Jackson crossed-cylinder vectors, J0 and J45. Right-eye results are reported. RESULTS: The mean spherical equivalent refraction measured by the Canon R-1 was -2.44 D. Measurements from the Grand Seiko were more hyperopic (mean M, -2.01 D), whereas those from the Nidek were more myopic (mean M, -2.66 D). Correlation of M for each pair of autorefractors was 0.99. For J0, the Canon was more minus than the other two instruments by 0.15 D compared with the Nidek and 0.13 D with the Grand Seiko, and on this component, the correlation of the Canon with each of the other two was 0.87. Mean J0 values for the Nidek and Grand Seiko were similar, 0.05 D and 0.03 D, respectively, and mean J45 values were 0.04 D for both instruments. Correlations were 0.97 between these two autorefractors for each of the two components. J45 measured by the Canon was more positive than the other two by 0.06 D. For J45, the correlation of the Canon with the Grand Seiko was 0.40 and with the Nidek was 0.38. In 92% of the eyes, the absolute difference in cylinder power between the Grand Seiko and the Nidek was < or =0.25 D. Only 42% of the differences between the Canon and the Grand Seiko and 40% of the differences between the Canon and the Nidek were this small. CONCLUSIONS: The Canon provided more myopic readings than the Grand Seiko and more hyperopic readings than the Nidek. The Canon measured more astigmatism that did not correlate well with the other instruments, whereas measurements of astigmatism taken by the Nidek and the Grand Seiko showed good agreement. If the Grand Seiko is used in place of the older Canon, the differences in spherical equivalent and astigmatism must be considered. PMID- 14747763 TI - The association between spherical and cylindrical component powers. PMID- 14747764 TI - The GLADEL multinational Latin American prospective inception cohort of 1,214 patients with systemic lupus erythematosus: ethnic and disease heterogeneity among "Hispanics". AB - Clinical and laboratory manifestations and outcome of systemic lupus erythematosus (SLE) may vary in different populations. A prospective multinational inception cohort should prove useful in identifying the influence of ethnicity on the clinical characteristics of SLE. We therefore analyzed clinical, laboratory, and prognostic variables in Latin American SLE patients with disease of recent onset who were entered into a prospective cohort, and compared these variables in the cohort's 3 major ethnic groups. Thirty-four centers from 9 Latin American countries participated by randomly incorporating SLE patients within 2 years of diagnosis into a standardized database. Participating centers were selected for their expertise in diagnosing and managing SLE. We were then able to evaluate prospectively socioeconomic variables, ethnicity, type of medical care, clinical and laboratory features, disease activity, damage, and mortality at each site. A coordinating center controlled the quality of the information submitted. Of the 1,214 SLE patients included in the cohort, 537 were mestizos, 507 were white, and 152 were African Latin American (ALA). (There were also small numbers of pure Amerindian and oriental individuals.) Significant differences were found between them in socioeconomic characteristics, type of care, and level of education favoring whites. Mestizos and ALA were younger at onset. Delay to diagnosis and disease duration was shorter in ALA. Fever was more frequent in whites; discoid lesions in ALA; renal disease and lymphopenia in mestizos and ALA. Although we found differences in background variables between ethnic groups from different countries, mestizos from 2 distant countries (Argentina and Mexico) were clinically akin and showed similar differences to whites. Mortality was associated with lower education, poor medical coverage, and shorter follow-up. In an exploratory model nonwhite ethnicity was associated with renal disease and lymphopenia, damage, and cumulative American College of Rheumatology criteria. These differences in clinical, prognostic, socioeconomic, educational, and access to medical care features in Latin American lupus patients of 3 major ethnic groups from 9 different countries may have an impact on the patients' disease. "Hispanics," as they have come to be generically termed on the basis of language, actually constitute a markedly heterogeneous group of subjects. PMID- 14747765 TI - Clinical features and metabolic and autoimmune derangements in acquired partial lipodystrophy: report of 35 cases and review of the literature. AB - We describe clinical features, body fat distribution, and prevalence of metabolic abnormalities in 35 patients with acquired partial lipodystrophy (APL) seen by us over 8 years, and also review 220 cases of APL described in the literature. Based on the review and our experience, we propose that the essential diagnostic criterion for APL is the gradual onset of bilaterally symmetrical loss of subcutaneous fat from the face, neck, upper extremities, thorax, and abdomen, in the "cephalocaudal" sequence, sparing the lower extremities. Analysis of the pooled data revealed that female patients were affected approximately 4 times more often than males. The median age of the onset of lipodystrophy was 7 years. Several autoimmune diseases, in particular systemic lupus erythematosus and dermatomyositis, were associated with APL. The prevalence rates of diabetes mellitus and impaired glucose tolerance were 6.7% and 8.9%, respectively. Approximately 83% of APL patients had low complement (C) 3 levels and the presence of polyclonal immunoglobulin C3 nephritic factor. Twenty-two percent of patients developed membranoproliferative glomerulonephritis (MPGN) after a median of approximately 8 years following the onset of lipodystrophy. Compared with patients without renal disease, those with MPGN had earlier age of onset of lipodystrophy (12.6 +/- 10.3 yr vs 7.7 +/- 4.4 yr, respectively; p < 0.001) and a higher prevalence of C3 hypocomplementemia (78% vs 95%, respectively; p = 0.02). The pathogenesis of fat loss and MPGN in patients with APL remains unclear, but activation of an alternate complement pathway has been implicated. Treating the cosmetic disfigurement by surgical procedures has yielded inconsistent results. The use of thiazolidinediones to treat fat loss in patients with APL remains anecdotal. Prognosis is mainly determined by renal insufficiency due to MPGN. PMID- 14747766 TI - Long-term survival of patients with idiopathic inflammatory myopathies according to clinical features: a longitudinal study of 162 cases. AB - The idiopathic inflammatory myopathies are characterized by chronic muscle inflammation and involvement of internal organs, which contribute considerably to the morbidity and mortality of the disease. We conducted the current study to determine the survival data for patients with idiopathic inflammatory myopathies according to the presence of extramuscular clinical manifestations. We also determined the cumulative survival probability and the long-term prognosis and analyzed the causes of death at a single clinical immunology center.A survival analysis was performed using data for 162 patients diagnosed between 1976 and 1997 according to Bohan and Peter's criteria. Patients were followed up for a minimum of 5 years (median, 101.5 mo) or to date of death. Cumulative survival probability was calculated by the Kaplan-Meier method. The influence of extraskeletal and extramuscular involvement was analyzed as prognostic factors for death by Cox proportional hazards survival model. Eighteen disease-specific deaths occurred; pulmonary and cardiac complications were the most frequent causes of death. Global survival rates were 95%, 92%, and 89% for 1, 5, and 10 years, respectively. Analysis for clinicopathologic subgroups revealed that cancer-associated myositis had the worst prognosis, while juvenile and overlap myositis had the best prognosis. Five- and 10-year survival rates were 94.2% and 89.4% for patients with primary polymyositis and 90.1% and 86.4% for primary dermatomyositis patients, respectively. In the whole group of patients with idiopathic inflammatory myopathy, cardiac (p < 0.01) and respiratory muscle involvement (p = 0.045) were significant prognostic factors for death. In the group of patients with primary polymyositis/dermatomyositis, cardiac involvement was the main prognostic factor for death (p < 0.01). Myositis patients described in this study have higher survival rates than reported previously worldwide. We examine the reasons for the differences between the data in the current study and the available survival data in the relevant literature. PMID- 14747767 TI - Multiple endocrine neoplasia type 1 and Zollinger-Ellison syndrome: a prospective study of 107 cases and comparison with 1009 cases from the literature. AB - In patients with multiple endocrine neoplasia type 1 (MEN1), the most common functional pancreatic endocrine tumor (PET) syndrome is Zollinger-Ellison syndrome (ZES). ZES has been well studied in its sporadic form (that is, without MEN1); however, there are limited data on patients with MEN1 and ZES (MEN1/ZES), and the long-term natural history is largely unknown. To address this issue we report the results of a prospective long-term National Institutes of Health (NIH) study of 107 MEN1/ZES patients and compare our results with those of 1009 MEN1/ZES patients in 278 case reports and small series in the literature. Patients were clinically, radiologically, and biochemically evaluated yearly for all MEN1 manifestations (mean follow-up, 10 yr; range, 0.1-31 yr). Compared with patients from the literature, the NIH MEN1/ZES patients more frequently had pituitary (60%) and adrenal (45%) disease and carcinoid tumors (30%), but had equal frequency of hyperparathyroidism (94%), thyroid disease (6%), or lipomas (5%). Twenty-five percent of both the NIH and the literature patients lacked a family history of MEN1; ZES was the initial clinical manifestation of MEN1 in 40%. ZES onset preceded the diagnosis of hyperparathyroidism in 45%. However, ZES was rarely (8%) the only initial manifestation of MEN1 if careful testing was done. ZES occurred before age 40 years in 50%-60% of the current patients, in contrast to older studies. The diagnosis of ZES is delayed 3-5 years from its onset and is delayed as long as in sporadic ZES cases. Pituitary disease and carcinoid tumors (gastric > bronchial, thymic) are more frequent than generally reported, whereas a second functional PET is uncommon. In patients with MEN1/ZES without a family history of MEN1, the MEN1 manifestations are not as severe. This study shows that MEN1/ZES patients differ in many aspects from those commonly reported in older studies involving few MEN1/ZES patients. In this study we have identified a number of important clinical and laboratory features of MEN1/ZES that were not previously appreciated, which should contribute to earlier diagnosis and improve both short- and long-term management. PMID- 14747768 TI - [Anti-cholesterol agents, new therapeutic approaches]. AB - Statins and fibrates constitute the two major families of lipid-lowering agents. Statins are widely used for the treatment of pure hypercholesterolaemia while fibrates are used for the treatment of hypertriglyceridemia. Both drugs are also used for the treatment of mixed dyslipidemia. Some fibrates efficiently lower serum LDL-cholesterol. Statins inhibit HMG-CoA reductase and decrease cellular cholesterol synthesis. The resulting lower intracellular cholesterol concentration induces the activation of SREBP thus inducing the over expression and transcription of the LDL receptor gene. This over expression of the LDL receptor in the liver increases the clearance of circulating LDL thus decreasing the LDL-cholesterol plasma levels. The effects of fibrates on lipid metabolism are entirely due to their capacity to activate PPAR-alpha and to induce the over expression of genes containing a PPRE in their promoter. Fibrates decrease triglyceride concentrations by increasing the beta-oxidation of fatty acids in the liver and by decreasing triglyceride-VLDL synthesis. Fibrates also decrease triglycerides by increasing the hydolysys of triglycerides in chylomicron and VLDL through their capacity to increase and to decrease the lipoprotein lipase and the apo C-III transcription, respectively. Fibrates could decrease triglycerides partly by inducing apo A-V over-expression. These molecules increase HDL-cholesterol by increasing apo A-I and apo A-II transcription. Therefore the mechanisms of action of statins and fibrates depend on their capacity to modulate the expression of genes controlling lipoprotein metabolism. PMID- 14747769 TI - [Electron transfer reactions: from electrons to drugs]. AB - Electron transfer reactions have been at the heart of great advances in organic and inorganic chemistry and biology since the early work in 1954 by H. Taube (Nobel Chemistry Prize in 1983) and R. Marcus (Nobel Chemistry Prize in 1991). In organic chemistry, a new kind of chain substitution reaction with paramagnetic species was defined and identified as S(RN)1 (substitution, radial-nucleophilic, unimolecular). In our laboratory, choosing more or less complex molecules which possess electron-withdrawing groups and correctly disposed alkylation sites has enabled us to extend this concept to the heterocyclic (S(RN)1 HET) and quinonic (S(RN)1 QUI) series. These studies led us to prepare, under mild operating conditions, highly branched molecules hard to obtain via other pathways and which possess high pharmacological activity in various fields. We have discovered new reaction mechanisms such as LD-S(RN)1 (long-distance S(RN)1) E(RC)1 (elimination radical chain, unimolecular), bis-S(RN)1 and poly-S(RN)1. Moreover, we have developed a new technology using microwaves which increases the interest of electron transfer reactions in medicinal chemistry for drugs synthesis. PMID- 14747770 TI - [Systemic insecticides: new risk for pollinator insects]. AB - Imidacloprid, a new systemic insecticide used as seed-dressing, has been widely used in France since 1994. Its application mode and its efficiency allow a significant reduction in comparison with the usual quantity of chemicals used during pulverising treatment. But the insecticide imidacloprid is suspected to have harmful effects on the pollinators as many bees have died since its introduction. Recent studies have shown that imidacloprid has chronic and sub lethal toxicities at levels of micro g/kg or less. It was therefore necessary to detect imidacloprid at these levels in soils, plants, flowers, and pollens. With this aim, we characterised the bio-availability of imidacloprid in the environment using a new quantitative analytical method, as a basis for the evaluation of the risk for bees. PMID- 14747771 TI - [Feasibility study of semi-solid matrices of theophylline-containing polyethyleneglycols]. AB - Pharmaceutical manufacturing of semisolid matrices a past meeting two requirements: sufficient fluidity and sufficient viscosity. Ideally, the paste should have strong viscosity associated with a high dissolution rate. We studied the correlation between the viscosity and dissolution rate of polyethylene glycol (PEG) semisolid matrices (SSM) enclosing theophylline as a tracer. Nine formulations containing PEG of different molecular masses were studied. This work revealed the singularity of PEG binary mixtures. For pharmaceutical manufacturing, the conclusion of this work is that PEG binary mixtures enable circumventing the generally verified constraint: strong viscosity involves low dissolution ability. PMID- 14747772 TI - [Immunosuppressive effects of a new phenothiazine derivative]. AB - An original phenothiazine, CPTZ, was tested for its effects on the mouse immune system. Serum opsonic capacity, phagocyte and bactericidal activity of peritoneal macrophages, counts of splenic cells forming hemolysis plaques, and the number of survivors after experimental infection were recorded. The effects observed were compared with those produced by levamisole (a non-selective immunomodulator) and indometacin (an antiinflammatory drug with selective immunomodulator properties). The effects of CPTZ might be useful for the development of a new class of immunosuppressor drugs. PMID- 14747774 TI - [Drug prescription typology in primary healthcare centers in Tunisian Sahel]. AB - The objective of this survey was to determine the typology of prescription drugs ordered in primary healthcare centers of the Tunisian Sahel. We conducted a cross sectional descriptive survey to analyze 2586 medical orders written by 20 general practitioners working in the public sector of the Sousse sanitary region. The sample included 5628 types of prescription for 209 pharmaceutical agents. More than half of the prescribed drugs (56%) belonged to two therapeutic families: anti-infectious agents and antalgesic-antiinflammatory agents. The 30 most prescribed drugs in general practice accounted for 80% of the prescription lines for agents belonging to 10 therapeutic families. In this "Top 30" list, we found seven antibiotics and four cardiology medicines. The prescription profile in primary healthcare in the Tunisian Sahel shows widespread use of drugs and reflects the diversity of the morbidity encountered in the general medicine setting which corresponds to an advanced "epidemiological transition" in this geographical area. The development of a "Top 30" list could be the starting point for more efficient pharmaceutical management and for more relevant education in therapeutics. PMID- 14747773 TI - [Action mechanism of anticonvulsant and anti-immobility (forced swim) effects of 3', 4'-dihydro-N, N-dimethylspiro-[9H-fluorene-9, 2' (5'H) furane]-3'-methanamine (AE37F)]. AB - AE37F, a new aminotetrahydrofuranic derivative, exhibited, at 10-30 mg/Kg (po) or 1-10 mg/Kg (ip), antagonism of tonic convulsions, induced by pentetrazole (130 mg/Kg, ip), and of forced swin immobility, in mice. At these doses AE37F induced a considerable (100-250%) increase of serotonin (5-HT) and its main metabolite, 5 hydroxyindolacetic acid (5-HIAA), in the rat nucleus reticularis pontis oralis (NRPO), antagonized by amantadine, which also increased 5-HT and 5-HIAA levels in the NRPO. It is suggested: a) that the anti-immobility effect of AE37F is related to its antimuscarinic properties, b) that the rate of 5-HT release in the NRPO, calculated here by a new approach (from the 5-HT and 5-HIAA brain levels) is increased by AE37F and decreased by amantadine, in the NRPO, c) that the anti convulsant action, observed with AE37F, could be related to a NMDA-sigma mediated stimulation of serotoninergic, GABAergic and glycinergic brain neurones, antagonized by the NMDA-sigma inhibition induced by amantadine. PMID- 14747775 TI - Primary prevention of type 2 diabetes. PMID- 14747776 TI - Prevalence of antithyroid microsomal antibody in thyroid patients of endemic goitre area. AB - The region of greater Mymensingh known for iodine endemicity, recently came under iodine supplementation as a result of mandatory universal iodination of salt program. Autoimmune thyroid diseases (AITD) are among the most common human autoimmune disorders & presence of autoantibodies to the microsomal antigen (AntiMCAb) is a hallmark of disease activity. Both iodine deficiency & iodine supplementation precipitate increase rate of autoimmunity to the thyroid gland. Study was undertaken to determine prevalence of AntiMCAb positive cases among patients with various thyroid diseases. High resolution ultrasound (HRUS), serum thyroid hormone assays & scintiscan were used to classify the thyroid patients into 8 categories. 221 patients were studied during the stipulated period of 3 months. Male patients were 60 & female patients were 161. Age ranged from 11 to 65 years with median age 29.4 years. AntiMCAb test were done with radioimmunoassay (RIA). 126 patients had antimicrosomal antibody (57.01%). All form of hypothyroid (atrophic, goitrous, Hashimoto's) have very high rate of AntiMCAb positive cases. Highest 89.28% were seen in patients showing feature of Hashimoto's thyroiditis or generalized feature of AITD in HRUS with hypothyroidism, followed had 61.29% positive cases, However, antithyroid antibody was found in all form of thyroid disorders. Nodular goiter had 21.73% antiMCAb positive cases. AntiMCAb found positive at the rate of 33.33% in euthyroid patients with HRUS feature of AITD & diffuse euthyroid goiter, 40% in subclinical hypothyroid, 40% in subclinical hyperthyroid. Female rated higher in range of antimicrosomal antibody positivism. 59% of all thyroid patients among female subjects were AntMCAb positive, where as 51.67% male thyroid patients were positive. Highest number of positive cases found in the 30-35 age group. No definite pattern, however, was observed among age distribution. 20 age matched sample from patients unsuspected of thyroid disease shows 10% AntiMCAb positive compared to 73.33% of the same among same age group of thyroid patients. Frank Hashimoto's thyroiditis with positive antiMCAb and hypothyroidism were all detected by HRUS. PMID- 14747777 TI - Coronary artery disease in young adults - angiographic profile. AB - Angiographic profile of fifty young patients of coronary artery disease aged 40 or under were analysed and compared with those of fifty older patients. Mean age of younger and older group was 36.34 (range 28 to 40 years) and 55.28 (44-74 years) years respectively and most of the patients were male in both the group (92 Vs 94%). Older patients were more diabetes (40 Vs 24%) and hypertensive (38 Vs 60%) but the younger patients had more family history of premature coronary artery disease (50% Vs 24%). The incidence of smoking and dyslipidemia did not vary between the two groups. Older patients had more history of myocardial infarction (69 Vs 58%) but angina were more in young patients (42 Vs 31%). Coronary angiography revealed more number of multivessel disease in older patients (74 Vs 54%) but the younger patients had more normal coronary arteries and single vessel disease (46 vs 26%). Coronary athesclerosis was also extensive in older patients as revealed by the higher coronary score, more involvement of coronary segments, more number of diseased and diffusely involved coronary vessel in older patients. Older patients needed more revasalarization process (74 Vs 60%), more coronary bypass surgery (40 Vs 24%) and had more inoperable vessels (16% Vs 4%) than the younger patients. So the younger patients having less extensive coronary artery athesclerosis with better prognostic probability should be evaluated angiographically for further definitive management in the from of revascularization. PMID- 14747778 TI - Role of fine needle aspiration cytology in the diagnosis of pulmonary lesions. AB - Fine needle aspiration cytology (FNAC) is a rapid, reliable and cost-effective method in the diagnosis of lung lesions. Cells are collected from the lesions usually under fluoroscopic or CT-guidance for cytological examination. Due to lack of these highly sophisticated radiologic imaging techniques in our institute we performed this test under X-Ray film guidance. This study was carried out in 59 patients, who underwent FNAC of lung lesions in the department of Pathology, Mymensingh Medical College, Bangladesh to find out the specimen adequacy and frequency of both benign and malignant lesions of the lungs in our areas. Out of 59 cases 54 were males and 5 were females with age ranged from 20 to 82 years. Adequate samples were obtained in 50 (84.75%) cases of which 35 (70%) were malignant, 10 (20%) were benign and 5 (10%) were suspicious for malignancy. Malignant lesions comprised of squamous cell carcinoma in 20 (57.14%), adenocarcinoma in 1(2.86%), large cell carcinoma in 8 (22.86%) and small cell carcinoma in 6 (17.14%) cases. All the benign lesions were inflammatory in nature, which included abscess in 6 (60%), tuberculosis in 3 (30%) and chronic non-specific inflammation in 1 (10%) case. Majority of the malignant lung lesions were encountered in the 6th decade of life. As the method is simple, rapid, cost effective and without major complications, so it can be practiced in any centres in our country where specialised radiologists and pathologists are available. PMID- 14747779 TI - Knowledge and attitude of faculty members on problem based learning. AB - An intervention study was carried out in Mymensingh Medical College during the second week of January, 2003 to compare knowledge and attitude of the faculty members of Mymensingh Medical College on PBL before and after exposure to a symposium. Pretest was done among 48 faculty members before and 45 faculty members after exposure to the symposium using a self-administered questionnaire. Before the symposium about 17% of faculty members had a sound knowledge on PBL, the figure rose significantly to about 61% after exposure to the symposium (P < 0.001). About 73 % of faculty members felt that PBL is effective in problem solving after exposure to the symposium, as compared to about 60% who felt that PBL is effective in problem solving before the symposium (P < 0.01). About 69% of faculty members stated that PBL is better than traditional ward teaching after exposure to the symposium, as compared to 51.1% who stated that PBL is better than traditional ward teaching before the symposium (P < 0.001). About 69% of faculty members stated that PBL enhances self-directed learning after exposure to the symposium, as compared to about 52% who stated PBL enhances self-directed learning before the symposium(P <0.005). About 64% of faculty members affirmed that they will welcome PBL in clinical teaching after exposure to the symposium, as compared to 62.5% of faculty members who would welcome PBL in clinical teaching before the symposium (P < 0.01). A total of 59% of faculty members asserted that they will recommend PBL to be included in Undergraduate Medical Curriculum after exposure to the symposium, as compared to 60.3% who would recommend PBL in Undergraduate Medical Curriculum before the symposium. About 51 % of faculty members agreed that practicing PBL will help students after graduation to continue independent learning before the symposium, as compared to 60% who agreed that practicing PBL will help students after graduation after exposure to the symposium (P < 0.05). It can be concluded that exposure to a symposium centered on PBL can improve knowledge and attitude of faculty members on PBL positively & significantly. PMID- 14747780 TI - Ultrasound guided fine needle aspiration cytology for diagnosis of mass lesions of liver. AB - The therapeutic and prognostic evaluation of malignant neoplasia of liver depends mostly on morphologic diagnosis. Ultrasound guided fine needle aspiration cytology (FNAC) of liver has been proved to be a rapid, reliable and cost effective method for cytological diagnosis of hepatic mass lesions. This study was undertaken to find out the frequency of different benign and malignant space occupying lesions (SOL) of liver from patients attending for FNAC in Mymensingh. Ultrasound guided FNAC was performed on 108 patients with hepatic mass lesions from September 2, 2001 to August 19, 2003. There were 67 (62.0%) males and 41 (37.96%) females with a mean age 53 year (SD +/- 14) ranging from 2 to 83 years. Samples were adequate in 101 (93.5%) and inadequate in 7 (6.5%). Out of 101 adequate samples only one was benign (liver abscess) and 100 were malignant. Of the malignant lesions hepatoblastoma was in 1 (1%), hepatocellular carcinomas in 8 (8%), metastatic adenocarcinomas in 73 (73%) and unclassified malignancies were in 18 (18%). Most of the hepatocellular carcinomas occurred in males (7 in 8). The frequency of metastatic adenocarcinoma was more in males (39 vs. 34) but the difference is not significant (p > 0.05). Ultrasound guided FNAC of liver is a rapid, reliable and cost-effective diagnostic method. It can be practice in any centres where ultrasound facility and specialist pathologists are available. PMID- 14747781 TI - Effect of prolonged use of injectable hormonal contraceptives on blood pressure and body weight. AB - The study was undertaken to determine the effect of prolonged use of injectable hormonal contraceptive on blood pressure and body weight in young women. Two hundred volunteers were selected for the purpose. Of them, one hundred and forty were taking injectable hormonal contraceptive, DMPA for 3 to 5 years uninterruptedly. Rest forty subjects served as control using no contraceptive steroid. Blood pressure, systolic as well as diastolic, was measured by sphygmomanometer. Body weight was measured by weighing machine. It was observed that there were insignificant (P>0.05) elevations of systolic and diastolic blood pressure from DMPA use. But body weight of the experimental subjects was significantly (P <0.05) increased in comparison to that of the control. PMID- 14747782 TI - Studies on clinical pattern of glomerulonephritis. AB - A prospective study of primary glomerulonephritis covering two and half years period was done at Dhaka Medical College Hospital & IPGMR, Dhaka from June 90 to July 92. A total of 100 cases were included in this study. All of the patients were of 13 years and above, clinically diagnosed, and by laboratory investigations, the diagnoses were established. In this study, we have tried to find out age incidence, sex ratio, possible aetiological factors, common clinical presentation, biochemical parameters and histological typing of primary glomerulonephritis. PMID- 14747783 TI - Treatment of non union of humerus using G. A. Ilizarov technique. AB - The management of the non union of humerus is one of the most challenging problems that the surgeon confronts in his practice. The procedures traditionally used are: I.M. Nailing, interlocking, plating, transplantation of allograft. In our series, 36 cases with non union has been treated by G.A. Ilizarov technique. The age range were: 21-62 years with an average of 32 years. The initial treatment were done by DCP, rush nails & plates with screw fixation. The duration of treatment ranged from 5-11 months (average 8 months). With the application of Ilizarov fixator a good range of elbow & shoulder motion were achieved. The average follow up period was 5 years with a range of 1-8 years. Union was achieved in all the 36 cases. PMID- 14747784 TI - A study of functional endoscopic sinus surgery technique. AB - Endoscopic sinus surgery has become a widely accepted surgical procedure for the treatment of chronic inflammatory sinus diseases. In our study 100 patients were suffering from polyposis treated by functional endoscopic sinus surgery in MMCH. Out of 100 patients 53 patients were suffering from ethmoidal polyp and 47 patients were suffering from antrochoanal polyp between the ages 7 to 45 years. Among antrochoanal polyp 2 patients were found with inverted papilloma and came with recurrence. 40% of the patients came with a follow up visit up to 1 year. 7 out of 53 cases of ethmoidal patient came with recurrence requiring revision FESS. There was no history of patient suffering from asthma or aspirin intolerance. Overall success rate was observed in 91%. Four patients were found with periorbital haematoma and 5 with fat coming out from orbit due to injury of lamina papyraceae. PMID- 14747785 TI - Management of dengue by the WHO guided national guidelines. AB - A total of 90 adult patients of Dengue admitted in Sir Salimullah Medical College and Mitford Hospital, Dhaka during the period from July 2002 to December 2002 were studied to see the presentation, spectrum of disease and outcome of management. Majority (52%) of cases was in the age group 21-30 years. Out of 90 patients 74 patients (82.22%) were male and 16 (17.78%) were female. Maximum number of cases (43%) was found in the month of August. Most of the cases (77%) were from Dhaka City. Classical type of Dengue fever was the commonest (80%) variety. Thrombocytopenia was found in 80% cases. Platelet count came down below 10000/cumm only in 1.1% of the patients. The patients were diagnosed mainly by clinical criteria. Serological tests were done only in selected cases. Majority (80%) of the patients was managed by sponging, paracetamol and Oral Rehydration Salt (ORS). Blood transfusion was required in 18% cases and platelet concentrate was given only in 1.1% cases. There was no mortality in this study. PMID- 14747786 TI - Metabolic and stress responses of the body to trauma: produced by the laparoscopic and open cholecystectomy. AB - Laparoscopic cholecystectomy is regarded as gold standard therapy for symptomatic gall stone disease. This study was done to compare the metabolic and stress responses between open and lapraroscopic cholecystectomy and to evaluate their significance in postoperative recovery. Thirty patients with symptomatic gall stone disease were treated with open and laparoscopic cholecystectomy on elective basis (14 versus 16). Three samples of blood were collected from each patient to investigate serum cortisol, adrenaline, nor-adrenaline, C-reactive protein and blood glucose level. The mean age of patients was 41.86+/-10.13 blood glucose and stress hormones (cortisol, adrenaline, nor adrenaline) and C-reactive protein all were found significantly raised in the postoperative period in open cholecystectomy than laparoscopic cholecystectomy group. The postoperative recovery was also prolonged in the open group. The obvious clinical advantages of laparoscopic cholecystectomy over open cholecystectomy is mainly because of less metabolic and stress response. PMID- 14747787 TI - Rickets among children of a coastal area of Bangladesh. AB - Many children with rachitic deformities have been reported in southern coastal area of Bangladesh but the actual rate of prevalence was not known. A survey was conducted to determine the magnitude of rachitic problem among the children of Chakaria thana of Cox's Bazar district of Bangladesh. Nine hundred children between 1-15 years selected randomly from 30 villages of total 340 villages. Face to face interview of the parents was taken and the children were examined for evidences of rickets. Serum calcium, phosphorus, alkaline phosphatase (ALP) were estimated and radiology of limbs were done in all clinically suspected cases and in a control of every eighth child. Seventy eight children (8.7%) had physical features suggestive of rickets. Fifty eight (6.4%) children had 'clinical rickekts' (positive physical feature(s) but normal ALP and negative radiology), 12 (1.3%) children had 'biochemical rickets' (positive physical features and raised ALP but negative radiology) and 8 (0.9%) children had 'confirmed rickets' (positive physical features, raised ALP and positive radiology). Out of 78 children with rachitic feature(s), Pectus carinatum was found as the most common clinical feature in 26 (33.3%) children followed by genu valgum in 23 (29.4%) cases. Twenty two normal children (2.2%) had raised level of ALP (>300U/L). The prevalence of rickets is high in children of Chakaria and further study is needed to find out the exact aetiology of rickets in children there. PMID- 14747788 TI - Seroprevalence of dengu infections amongst the children. AB - Serosurvillance of 650 children of clinically suspected dengue infection based on simple laboratory tests were evaluated to find out the early indicator for diagnosis of dengue infection. The study was conducted among the patients attending the Sir Salimullah Medical College (SSMC) and Mitford Hospital and Popular Diagnostic Center, Dhaka, during July 2002 to September 2002. Of these 650 suspected dengue cases, 294 were in the age group 1-5 years, 206 were in age group 6-10 years and 150 cases in age group 11-15 years, of which Seropositivity was found in 78 (26.53%), 60 (24.12%) and 65 (43.33%) cases respectively. Primary dengue IgM was positive in 123 (60.59%) cases and secondary dengue (IgG & IgM combined or IgG alone) was positive in 80 (39.41%) cases. Leucopenia was found in 90 (75%) cases and Thrombocytopenia was found in 95 (79.16%) cases in primary dengue cases. Similarly in secondary dengue cases TWBC and PLT was decreased in 60 (72.29%) and 65 (78.15%) cases respectively. Haemoconcentration was found in 40 (36.67%) cases of primary dengue and 13 (18%) cases in secondary dengue. Month wise distribution of dengue seropositivity shows that 10 (8.53%) cases was found in July, 175 (50.10%) cases in August and 78 (10.00%) cases in September. This study reveals that seropositivity is highest (43.33%), in the age group 11-15 years and incidence of dengue increased in the month of August. Leucopenia (WBC or =50% of ED visits for migraine) and "other frequent ED visitors." RESULTS: There were 2,158,291 ED visitors, of whom 6,839 were frequent ED visitors. These individuals accounted for 3.5% of all ED visits. Among frequent ED visitors, 478 were frequent migraineurs. The proportion of patients who were female was 79% among frequent migraineurs, 53% among other frequent ED visitors, and 50% among patients who were not frequent ED visitors. The proportions between ages 30 and 54 years for these 3 groups were 79%, 43%, and 31%, respectively. Average annual visits to EDs were 26, 18, and 1.7, respectively, and average visits to family physicians were 40, 20, and 5.9, respectively. Despite heavy health care use, frequent migraineurs sought 83% of their ED care from their most frequently visited ED and 71% of their primary care from their main family physician. CONCLUSION: Frequent migraineurs are predominantly women aged 30 to 54 years and with a particularly intense use of health care services. Management strategies may require targeted interventions for these individuals. Because these patients seek most of their care from 1 main ED and 1 principal primary care physician, coordination of care may be easier than expected. PMID- 14747815 TI - Postpartum blindness: two cases. AB - We present 2 cases, one eclamptic patient and one noneclamptic patient, of headache, cortical blindness, and seizures. Both patients demonstrated findings consistent with posterior leukoencephalopathy syndrome. Posterior leukoencephalopathy syndrome is a rapidly evolving neurologic condition that is characterized by headache, nausea and vomiting, seizures, visual disturbances, altered sensorium, and occasionally focal neurologic deficits. Posterior leukoencephalopathy syndrome can be triggered by numerous conditions, including preeclampsia-eclampsia, and can be seen in the postpartum period. It is characterized predominately by white matter vasogenic edema of the occipital and posterior parietal lobes. This condition can be difficult to differentiate clinically from cerebral ischemia, and magnetic resonance imaging with diffusion weighted imaging and apparent diffusion coefficient are needed to do so. In most cases of posterior leukoencephalopathy syndrome, the prognosis is excellent, with full resolution of symptoms. PMID- 14747816 TI - Bispectral index monitoring quantifies depth of sedation during emergency department procedural sedation and analgesia in children. AB - STUDY OBJECTIVE: The bispectral index monitor uses processed electroencephalogram signals to measure sedation depth on a unitless scale from 0 to 100 (0, coma; 40 to 60, general anesthesia; 60 to 90, sedated; 100, awake). It has been validated in the operating room as an objective measure of sedation depth with nondissociative general anesthesia; however, its usefulness in the pediatric emergency department (ED) for procedural sedation and analgesia has not been established. We determine the ability of the bispectral index to monitor depth of nondissociative procedural sedation and analgesia in children. METHODS: This was an observational study conducted in a children's hospital ED. Procedural sedation and analgesia was performed in the standard manner, with the addition of bispectral index monitoring and simultaneous clinical sedation scoring (modified Ramsay Sedation Scale [range 1 to 8; 1=alert, 8=unresponsive]). Paired bispectral index and Ramsay Sedation Scale scores were assigned every 5 minutes during the sedation. Ramsay Sedation Scale scores were assigned by a single study investigator blinded to the bispectral index score. An emergency physician independently administered all medications for procedural sedation and analgesia. The correlation between the paired bispectral index/Ramsay Sedation Scale scores was determined by using a repeated-measures regression analysis. Receiver operator characteristic (ROC) curves were constructed to determine the ability of the bispectral index to discriminate various thresholds of sedation depth. RESULTS: A convenience sample of 20 patients was enrolled, providing 217 paired bispectral index/Ramsay Sedation Scale measurements. Median age was 4.6 years (range 0.4 to 16.7 years). Fourteen patients received midazolam with fentanyl; the remainder received pentobarbital. Bispectral index scores ranged from 40 to 98 (mean 81.6+/-16.1). Ramsay Sedation Scale scores ranged from 1 to 8 (median 3; interquartile range 2 to 4). The simple Pearson correlation between paired bispectral index and Ramsay Sedation Scale scores was -0.78 (95% confidence interval [CI] -0.83 to -0.72; P<.001). After adjustment for the nonindependence of intrapatient data with bivariate repeated-measures analysis, the correlation was -0.67 (95% CI -0.90 to -0.43; P<.001). The linear regression coefficient between bispectral index and Ramsay Sedation Scale scores was estimated to be between -5.7 and -12.7. ROC curve analysis demonstrated moderate to high discriminatory power of bispectral index scores in predicting level of sedation throughout the sedation continuum, with areas under the curve at least 0.87 for all Ramsay Sedation Scale score thresholds. Bispectral index scores between 60 and 90 predicted with moderate accuracy traditional clinical levels of sedation typically encountered during procedural sedation and analgesia in the pediatric ED. CONCLUSION: Bispectral index monitoring correlated with clinical sedation scores and may serve as a useful, objective adjunct in quantifying depth of nondissociative procedural sedation and analgesia in children. PMID- 14747818 TI - Mass casualty terrorist bombings: a comparison of outcomes by bombing type. AB - STUDY OBJECTIVE: We compared the epidemiologic outcomes of terrorist bombings that produced 30 or more casualties and resulted in immediate structural collapse, occurred within a confined space, or occurred in open air. METHODS: We identified eligible studies of bombings through a MEDLINE search of articles published between 1966 and August 2002 and a manual search of published references. Pooled and median rates of mortality, immediately injured survival, emergency department use, hospitalization, and injury were determined for each bombing type. RESULTS: We found 35 eligible articles describing 29 terrorist bombings, collectively producing 8,364 casualties, 903 immediate deaths, and 7,461 immediately surviving injured. Pooled immediate mortality rates were structural collapse 25% (95% confidence interval [CI] 6% to 44%), confined space 8% (95% CI 1% to 14%), and open air 4% (95% CI 0% to 9%). Biphasic distributions of mortality were identified in all bombing types. Pooled hospitalization rates were structural collapse 25% (95% CI 6% to 44%), confined space 36% (95% CI 27% to 46%), and open air 15% (95% CI 5% to 26%). Unique patterns of injury rates were found in all bombing types. CONCLUSION: Patterns of injury and health care system use vary with the type of terrorist bombing. PMID- 14747817 TI - Treatment of pediatric migraine headaches: a randomized, double-blind trial of prochlorperazine versus ketorolac. AB - STUDY OBJECTIVE: We compare the effectiveness of intravenous ketorolac and intravenous prochlorperazine in the treatment of pediatric migraine headaches. METHODS: We performed a prospective, randomized, double-blind clinical trial in 2 pediatric emergency departments (EDs) within children's hospitals. Children aged 5 to 18 years presenting to the ED with migraine headaches were eligible for the study. Contraindications to either medication or the inability to complete the pain score resulted in exclusion. Children were randomized to receive intravenous ketorolac (0.5 mg/kg; maximum 30 mg) or intravenous prochlorperazine (0.15 mg/kg; maximum 10 mg). All children also received a normal saline solution bolus. Successful treatment was defined as a 50% or greater reduction in the Nine Faces Pain Scale score at 60 minutes. If a less than 50% improvement occurred by 60 minutes, the child received the other medication. Forty-eight-hour follow-up telephone calls were made to each family to assess recurrence and late side effects. RESULTS: Sixty-two children were enrolled: 33 initially received prochlorperazine, and 29 initially received ketorolac. By 60 minutes, 16 (55.2%) of 29 of those who received ketorolac and 28 (84.8%) of 33 of those who received prochlorperazine were successfully treated (difference=30%; 95% confidence interval [CI] 8% to 52%). Fifty-six (93.3%) of the 60 children who completed the study were successfully treated by the study's conclusion. Approximately 30% of each group had a recurrence of some headache symptoms. Only 2 children reported side effects, both mild and self-limited. CONCLUSION: In children, intravenous prochlorperazine is superior to intravenous ketorolac in the acute treatment of migraine headaches. PMID- 14747819 TI - National Highway Traffic Safety Administration (NHTSA) notes. PMID- 14747821 TI - Clinical policy: Critical issues in the evaluation of adult patients presenting to the emergency department with acute blunt abdominal trauma. PMID- 14747822 TI - Images in emergency medicine. Pulmonary embolus. PMID- 14747823 TI - Mental health status among ethnic Albanians seeking medical care in an emergency department two years after the war in Kosovo: a pilot project. AB - STUDY OBJECTIVE: The long-term psychological effects of war are under appreciated in clinical settings. Describing the postwar psychosocial burden on medical care can help direct public health interventions. We performed an emergency department (ED)-based assessment of the mental health status of ethnic Albanian patients 2 years after the North Atlantic Treaty Organization-led bombing of Serbia and Kosovo in 1999. METHODS: This study was conducted July 30, 2001, to August 30, 2001, in the ED of a hospital in Pristina, Kosovo. Investigators collected data through systematic sampling of every sixth nonacute ED patient presenting for care; 87.7% of patients agreed to participate. Respondents completed a structured questionnaire, including demographic characteristics, the Short Form-36, and the Harvard Trauma Questionnaire. RESULTS: All 306 respondents were ethnic Albanians; mean age was 39 years (SD 17.9 years). Of respondents, 58% had become refugees during the war. Two hundred ninety-six (97%) reported experiencing at least one traumatic event during the war; the average number of traumatic events encountered by participants was 6.6. Forty-three (14%) reported symptoms that met Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition criteria for posttraumatic stress disorder; mean Short Form-36 Mental Component Summary score was 42.1 (SD 12.5). Separate multivariable linear regression models confirmed our belief that older age, female sex, less than a high school education, and having experienced a greater number of traumatic events would be associated with more posttraumatic stress disorder symptoms and lower Mental Component Summary scores. CONCLUSION: Mental health problems among ED patients in Kosovo, particularly among specific vulnerable populations, are a significant public health concern 2 years after the conflict. PMID- 14747824 TI - Emergency medicine and international health: increasing impact by broadening the mandate. PMID- 14747826 TI - When once is enough. PMID- 14747825 TI - Phasic alterations in dopamine and serotonin release in striatum and prefrontal cortex in response to cocaine predictive cues in behaving rhesus macaques. AB - The ability of environmental cues associated with cocaine availability to cause relapse may result from conditioned activation of dopamine (DA) release. We examined this hypothesis in macaque monkeys by conducting microdialysis studies in animals during exposure to a cocaine predictive compound cue. In addition to studying DA release in mesolimbic and sensorimotor striatum, both DA and serotonin levels were determined in the prefrontal cortex (medial orbitofrontal and anterior cingulate). The compound cue employed visual, auditory, and olfactory components, and was salient to the animals as demonstrated by anticipatory lever pressing in the absence of cocaine. During a 10-min period of exposure prior to cocaine availability, there was no significant increase in striatal or cortical DA. The addition of a DA uptake inhibitor to the striatal perfusate to reduce the potential interference of neuronal uptake did not alter the results. In contrast to the lack of any change in striatal DA, a significant decrease in extracellular serotonin in the prefrontal cortex during the 10 min of cue exposure was observed. PMID- 14747828 TI - Recruitment order: a powerful neural ensemble code. PMID- 14747829 TI - Storing memories in dendritic channels. PMID- 14747830 TI - Taking sides in the nervous system with miRNA. PMID- 14747831 TI - Listening to your heart: interoceptive awareness as a gateway to feeling. PMID- 14747832 TI - Localized striatal delivery of GDNF as a treatment for Parkinson disease. PMID- 14747833 TI - A reanalysis of the indirect evidence for recombination in human mitochondrial DNA. AB - In an attempt to resolve the controversy about whether recombination occurs in human mtDNA, we have analysed three recently published data sets of complete mtDNA sequences along with 10 RFLP data sets. We have analysed the relationship between linkage disequilibrium (LD) and distance between sites under a variety of conditions using two measures of LD, r2 and /D'/. We find that there is a negative correlation between r2 and distance in the majority of data sets, but no overall trend for /D'/. Five out of six mtDNA sequence data sets show an excess of homoplasy, but this could be due to either recombination or hypervariable sites. Two additional recombination detection methods used, Geneconv and Maximum Chi-Square, showed nonsignificant results. The overall significance of these findings is hard to quantify because of nonindependence, but our results suggest a lack of evidence for recombination in human mtDNA. PMID- 14747834 TI - Analysis of the CARD15 variants R702W, G908R and L1007fs in Italian IBD patients. AB - CARD15 on chromosome 16 is the only IBD susceptibility gene identified among several mapped loci. Its recurrent variants R702W, G908R and L1007fs have shown significant association with Crohn's disease (CD), but not with ulcerative colitis (UC), in different Caucasian populations. We analysed these three variants in 184 CD and 92 UC Italian patients and in 177 healthy controls. L1007fs and G908R were independently associated with CD, while R702W showed a nonsignificant increase. After combining the three variants together, 32.6% of CD patients were positive vs 18.6% of the controls. The association was stronger for homozygotes and compound heterozygotes, OR 13.9 (1.8-108), and weaker but still significant for simple heterozygotes, OR 1.7 (1.0-2.9). An excess of homozygotes/compound heterozygotes also resulted from the comparison with Hardy Weinberg expectations. Phenotype-genotype correlations were analysed first by univariate logistic regression and then by multivariate analysis, the effect of CARD15 positivity being adjusted according to the status of smoking, familiarity and sex, so as to focus on the predictivity of genetic and environmental risk factors on the clinical phenotype. Significant risk estimates of the CARD15 genotype were obtained for stricturing vs inflammatory behaviour, OR 2.76 (1.2 6.3), and for penetrating behaviour, 2.59 (1.0-6.6), and marginally significant for ileal vs colic location, OR 3.0 (0.9-9.8). Our findings indicate that the association of the CARD15 genotype with behaviour and location of disease holds also for the Italian population. PMID- 14747835 TI - Advanced parental age in maternal uniparental disomy (UPD): implications for the mechanism of formation. AB - Uniparental disomy (UPD) describes the inheritance of a pair of chromosomes from only one parent. Meiotic nondisjunction followed by trisomy rescue is considered to be the major mechanism of formation. A literature search for cases with whole chromosome UPD other than UPD 15 was performed. Information on parental age was available in 111 cases with maternal UPD and in 34 cases with paternal UPD. In 52 out of 74 cases with maternal heterodisomy, information on the time of nondisjunction was also available. Around two-thirds of these cases were due to a maternal meiosis I error. Compared with the mean maternal age of 30.0 years in Bavarian mothers, in the year 2000 an advanced mean maternal age of 34.8 years was found in cases with maternal heterodisomy (n=74; P<0.0001). Almost no difference in the mean maternal age was observed between meiosis I errors (35.56 years; n=30) and meiosis II errors (35.78 years; n=14). The mean maternal age was 31.46 years in cases with maternal isodisomy and a normal karyotype (n=24), and the mean paternal age was 31.48 years in cases with paternal isodisomy (n=28). The various mean parental ages in heterodisomic and isodisomic cases are considered to reflect strongly the different mechanisms of formation: trisomy rescue or gamete complementation, which implies a meiotic nondisjunction in maternal heterodisomic UPD, and postzygotic somatic reduplication in cases with paternal and maternal isodisomic UPD. PMID- 14747836 TI - Ciliary neurotrophic factor (CNTF) genotype and body composition. AB - Exogenous ciliary neurotrophic factor (CNTF) administration causes significant weight loss in both humans and animal models, but the effects of endogenous CNTF and the CNTF null allele on body composition are not fully understood. A recent study in a European cohort demonstrated a significantly higher body weight and body mass index (BMI) in older males homozygous for the CNTF null allele (A/A genotype). We sought to replicate these findings in three cohorts: the Baltimore Longitudinal Study on Aging (BLSA) consisting of 422 adult men and women (19-90 years); the Study of Osteoporotic Risk in Men (STORM) consisting of 333 older men (50-84 years); and a third sample obtained by combining older males aged 59-73 years from the BLSA and STORM cohorts (n=286). In contrast to the European study, we were unable to detect a significant association between CNTF genotype and body weight in the BLSA (P=0.49), the STORM (P=0.28), or the combined samples (P=0.72). There was also no significant association observed between CNTF genotype and BMI in the BLSA (P=0.59), the STORM (P=0.34) or the combined (P=0.56) samples. In addition, we were unable to detect a significant association between CNTF genotype and total body fat (P=0.95) or fat-free mass (P=0.86) in the BLSA cohort. Our results do not support an effect of the CNTF null allele on body composition, contrary to previous findings. PMID- 14747837 TI - Pharmacological modulation of local feedback mechanisms as a therapeutic approach in breast cancer treatment. AB - Breast function and development are regulated by a network of local and systemic signals which can exert either stimulatory or inhibitory effects. Many of these signals are mediated by topically produced hormones and cytokines, which are both believed to be part of complex feedback loops. Over the last few years it has become increasingly clear that local feedback mechanisms also play an integral part in the growth and local invasion of malignant breast tumors. The intratumoral development of positive feedback loops can not only enhance the malignant phenotype and growth rate of tumor cells, but is also thought to stimulate the formation of vascular elements to ensure a sufficient supply of nutrients. In addition, the disruption of physiological negative feedback loops in breast tissue might result in the loss of cell cycle control and eventually lead to increased tumor growth and local breakdown of adjacent stroma. It is therefore not surprising that many of these loops involve interactions between tumor cells and their stromal environment, and are located at the site of tumor invasion. Since several of the potentially involved cytokines are already known and a number of specific inhibitors are now available, it is possible to interrupt pathological intercellular communication by selectively inhibiting intratumoral key feedback mechanisms. While the identification of each new intratumoral feedback loop can help us in our quest for for novel specific antineoplastic strategies, we now also know that some of the most commonly used endocrine therapies are probably so effective because they also interfere with local intercellular communication. PMID- 14747838 TI - Genes implicated in the pathogenesis of spinocerebellar ataxias. AB - The degenerative ataxias comprise a number of heterogeneous diseases, many of which are genetically determined. Loss of cerebellar Purkinje and brainstem neurons as well as degeneration of spinal pathways are the major morphological findings of most ataxias, but neuronal loss may also affect the basal ganglia and the retina. While the degenerative ataxias initially were classified on a neuropathological basis, more recent classifications focused on clinical hallmarks and the mode of inheritance, separating inherited, sporadic and symptomatic ataxias. Genetic linkage analysis and molecular genetic studies identified various genotypes and revealed genetic heterogeneity of the autosomal dominant ataxias (ADCA), which on the basis of the genotypes are now classified as spinocerebellar ataxias (SCA1-22). Based on pathogenesis these disorders fall into three discrete groups: the polyglutamine disorders, SCA1-3, 7 and 17; the channelopathies, SCA6 and episodic ataxia types 1 and 2 (EA1-2); and SCA8, 10 and 12, which result from repeat expansions outside the coding regions and reduce gene expression. The etiologies of SCAs 4, 5, 9, 11, 13-16, 19, 21 and 22 remain unknown as of today. The recent advances in the identification of the underlying gene defects of most of the inherited ataxias have opened new avenues to a better understanding of the molecular mechanisms leading to cellular dysfunction and cell death. PMID- 14747839 TI - Gastrointestinal-related adverse effects of COX-2 inhibitors. AB - Selective cyclooxygenase-2 (COX-2) inhibitors are used for the treatment of inflammation and pain while having the reported advantage of fewer upper gastrointestinal adverse effects compared to traditional nonsteroidal anti inflammatory drugs. Although fewer adverse effects occur, there is still a risk for developing upper gastrointestinal adverse effects. Clinical practitioners have increased concern regarding this risk. The belief that COX-2 inhibitors are safe for the gastrointestinal tract has been questioned. This has encouraged the proposal of several explanations on the mechanism of gastromucosal injury and healing relative to COX isoenzymes. These mechanisms are delineated in the following review, along with the gastrointestinal safety, risk factors, clinical and case studies, and cost effectiveness of the COX-2 inhibitors. PMID- 14747840 TI - Cardioprotective effects of thiazolidinediones, peroxisome proliferator-activated receptor-gamma agonists. AB - Greater myocardial injury in response to ischemia/ reperfusion (I/R) and increased incidence of congestive heart failure and death in noninsulin-dependent diabetes mellitus, or type 2 diabetes, patients has been clearly identified. Thiazolidinediones (TZDs), peroxisome proliferator-activated receptor-gamma (PPAR gamma) agonists, act as insulin sensitizers and are a novel class of oral antidiabetic drugs. An emerging body of evidence, mainly from preclinical studies, suggest that TZDs protect the heart from acute I/R injury and also might attenuate cardiac remodeling and heart failure. The mechanisms involved in this cardioprotection by TZDs are multi-factorial and not completely understood. These novel activities of TZDs could benefit type 2 diabetes patients and offer benefits beyond glycemic control. This new knowledge about the cardioprotective effects of TZDs is still limited, and more investigations, especially clinical studies, are required. PMID- 14747841 TI - Endometriosis: a genetic disease. AB - Endometriosis is a multifactorial disease affecting up to 15% of women of reproductive age. This condition is characterized by the presence and growth of endometrial cells outside the uterus. Susceptibility to endometriosis depends on complex interactions of immunologic, hormonal, environmental and genetic factors. Although familial inheritance plays a role, multiple candidate genes appear to be involved. New studies investigating the influence of genetic variants on endometriosis are published with increasing frequency. A number of technologies have emerged to facilitate progress in this field, including subtractive cDNA hybridization to identify secretory endometrial genes, DNA chip technology, differential display polymerase chain reaction, cytogenetics evaluation of endometriotic cells and tissues, and complementary methods in proteomics and informatics. One general approach to uncovering genes pivotal to endometriosis is to search systematically for perturbations in selective candidate genes or chromosomal regions using polymerase chain reaction. We describe here novel association studies on obvious candidates, including genes governing cancer susceptibility, hormone sensitivity or immunology. The assessment of mutations and polymorphisms may allow individualization of therapies as well as primary and secondary prevention strategies for endometriosis, aimed at high-risk populations. PMID- 14747842 TI - The Mirena levonorgestrel system. AB - The levonorgestrel intrauterine system is a safe, efficacious, long-term contraceptive device. Additionally, it results in a decrease in the volume of menstrual blood loss in women with normal periods and those with menorrhagia. It may also have a role in the treatment of other benign gynecological disorders, such as adenomyosis or endometriosis, and it has been advocated as a means of delivering progestogen to the endometrium as part of combined hormone replacement therapy. Despite the beneficial effect on menstrual volume, compliance is sometimes limited by breakthrough bleeding during the first months of use. The precise mechanism of this bleeding is unclear, but it may be related to a direct effect of levonorgestrel on endometrial vascular development. Detailed counseling is crucial to explain this anticipated side effect in order to reduce unnecessary discontinuation of treatment. PMID- 14747843 TI - The pharmaceutical industry and research in 2002 and beyond. AB - The success of the pharmaceutical industry will continue to depend on its ability to satisfy the clinical needs of established market economies. The number and quality of new drugs emerging from development pipelines seems likely to rise due to increased research and development budgets of the merged pharmaceutical companies, efficiencies across all facets of the development process, increasing use of new technologies and availability of new targets from the ongoing work on the role of human genes in disease pathways. In addition to the traditional small molecule drugs, the market for protein products, including monoclonal antibodies and therapeutic vaccines, is likely to expand as advances in recombinant and formulation technologies are made. Current work on relatively newer fields of pharmaceutical research, such as novel G-protein-coupled receptors, chemokines/cytokines, integrins and control of cell cycle regulation and signal transduction pathways (kinases, phosphatases and transcription factors) will lead to new drugs over the next decade. It is tempting to argue that a progressive fall in the number of new drugs in the last decade of the 20th century reflects the end of an era as companies struggle to identify any remaining quality products using old-style drug hunting practices. PMID- 14747844 TI - The biological basis of Hodgkin's lymphoma. AB - In the past several years our understandings for Hodgkin's lymphoma have significantly progressed, and we can now recognize two fundamental bases of Hodgkin's lymphoma: germinal center B cells as a cellular origin of Hodgkin and Reed-Sternberg (H-RS) cells and constitutively strong NF-kappaB activation as a biological base for H-RS cells. We can also define Hodgkin's lymphoma as being composed of H-RS cells with self-growth-promoting potential as malignant cells by constitutively strong NF-kappaB activation and surrounding reactive cells. Identification of molecules involved in constitutive and strong NF-kappaB activation in H-RS cells is important to understand the pathophysiology as well as transformation and developmental process of Hodgkin's lymphoma. Epstein-Barr virus latent membrane protein (LMP)-1, defective IkappaBalpha, IkappaB kinase activation and ligand-independent signaling by overexpressed CD30 have been clarified in the past several years. Involvement of JunB in overexpression of CD30 has been recently reported. Today, over a century and a half after the first report by Thomas Hodgkin, we at last obtained several keys to solving the mystery of Hodgkin's lymphoma on a biological basis. PMID- 14747846 TI - The role of Krs1 in cell cycle arrest. AB - The Krs1 (kinase responsive to stress 1) kinase has been proposed to mediate signals initiated by various forms of cellular or environmental stress for the process of growth arrest and apoptosis. The functional role of Krs1 in cell growth arrest may involve a constant, stress-independent proteolytic modification of the full-length kinase p63(Krs1), via caspase-like activity, to produce kinase active kinase fragment, p33(Krs1). Induction of the kinase activity of p33(Krs1) is closely correlated with cell growth arrest in the G(0)/G(1) phase due to serum starvation, contact inhibition or growth-arresting agents. Deactivation of p33(Krs1) is closely associated with cell entry into the cell cycle. In response to stress shock the conversion of p63(Krs1) to p33(Krs1) is enhanced, and the kinase activity of p33(Krs1) is additionally increased in quiescent cells that undergo apoptosis. Both mechanisms of proteolytic modification and protein phosphorylation are involved in a complex control of the kinase activity and function of p33(Krs1) in response to environmental changes leading to cell growth arrest in quiescence or quiescence-related apoptosis. Numerous studies have shown that signaling pathways in malignantly transformed cells are regulated differently from their counterpart pathways in normal counterpart cells in response to stress shock or anticancer agents. It is desirable to develop one type of anticancer agent to selectively growth-arrest normal counterpart cells in G(0)/G(1) phase of the cell cycle through the control of Krs1-involved signaling pathways. The success of arresting normal cells in a quiescent state will provide access for other types of anticancer treatments to induce cell death of cancerous cells, which are resistant to growth arrest in other phases of the cell cycle. PMID- 14747845 TI - Hedgehog signaling pathway as a target for therapeutic intervention in basal cell carcinoma. AB - Basal cell carcinomas (BCC) are slow-growing skin tumors that rarely metastasize but frequently recur, most often around the face, head and neck. Currently, surgery is the only treatment practice, which can be painful and leave scars. However, several years ago it was discovered that almost all forms of BCC result from mutations in a signaling pathway controlled by a protein called Hedgehog (Hh). Recently, a novel small-molecule drug candidate, CUR-61414, has been identified that blocks this pathway and could potentially be effective for the treatment of BCC. CUR-61414 was reported to prevent the proliferation and selectively induce the death of the tumor cells, while not harming adjacent normal skin cells in two different models of BCC. These findings directly demonstrate that the use of Hh inhibitors could be a valid novel therapeutic approach for treating BCC. PMID- 14747847 TI - Drugs to interfere with orexins (hypocretins). AB - Orexins (hypocretins) are bioactive peptides linking arousal, appetite and neuroendocrine-autonomic control. Dysfunction of the orexin system is associated with narcolepsy-cataplexy. Here, we review drugs interfering with orexins directly such as novel selective orexin receptor agonists and antagonists, as well as drugs interfering with orexins indirectly such as those used for the treatment of narcolepsy-cataplexy, and pharmacological targets within the complex network of endogenous neurohumoral signals integrated and relayed by the orexin system. These include amines, acetylcholine, purines, GABA and glutamate, as well as nutritional-metabolic, circadian-photic, immunological and neuroendocrine peptidergic influences. Basic and clinical evaluation of drugs interfering with the orexin system will lead to a better understanding of the molecular prerequisites that control behavioral state, stress responses, energy homeo- stasis and survival, and yield therapeutic advances for the treatment of narcolepsy and other disorders of sleep, eating, mood and memory. PMID- 14747848 TI - Molecule of the month. Ruboxistaurin mesilate hydrate. PMID- 14747849 TI - Highlights of the 11th Congress of the International Headache Society, held September 13-16, 2003, in Rome, Italy. Updating the headache classification system. AB - The International Headache Society held their 11th biennial congress in Rome, Italy, on September 13-16, 2003. The meeting featured the introduction of the second edition to the International Classification of Headache Disorders, the first update to the document since its presentation in 1988. This report discusses some of the different classifications of headache and their treatments as discussed at the congress. PMID- 14747850 TI - Traditional Chinese drug therapy. AB - More than 4,000 years old, traditional Chinese medicine continues to be widely practiced in China and in western countries. Traditional Chinese medicine teaches that good health is the result of harmony and balance between five basic elements: earth, water, fire, wood and metal. Also important to health are the two types of energy Yin and Yang, constituting a vital substance that circulates through the body. Drug therapy has been one of the means used in Chinese medicine to keep these elements and the flow of energy in balance. Many of the same herbs used thousands of years ago in China could be the source of new pharmaceuticals in Western medicine. PMID- 14747851 TI - Nucleic acid nanotechnology-towards Angstrom-scale engineering. AB - Nucleic acids and analogues are suitable building blocks for reliable self assembly of nanometer-sized two- or three-dimensional materials. In order to mimic or approach nature with respect to size and function, Angstrom-scale chemical engineering is emerging as pivotal for future developments. Efforts within nucleic acid nanotechnology will be focussed on generating rigid and stable low nanometer-sized structures carrying functionalities with predictable spatial positioning allowing, by encoded self-assembly, functional nucleic acid architectures to be built towards applications within the biological and material sciences. PMID- 14747852 TI - Synthesis and bio-assay of RCM-derived Bowman-Birk inhibitor analogues. AB - Bowman-Birk inhibitor analogues containing 2, 3 and 4-carbon analogues of the natural disulfide were synthesised via solid phase microwave-assisted RCM and found to have K(i) values against chymotrypsin in the low to sub-micromolar range, the best replacement for the disulfide arising from the linkage by RCM of two l-homoallylglycine residues. PMID- 14747853 TI - Palladium-catalyzed addition of disulfides and diselenides to alkynes under solvent free conditions. AB - An efficient methodology was developed for performing palladium-catalyzed E-E (E = S, Se) bond addition to alkynes under solvent free conditions. Compared to reaction in solvent significant enhancement of reaction rate, improved efficiency and remarkable catalyst stability were observed under solvent free conditions. The addition reactions were carried out with high stereoselectivity and yields in a short reaction time. PMID- 14747854 TI - Substrate-assisted antibody catalysis. AB - A new strategy in transition-state analog design is demonstrated to elicit catalytic antibodies. The strategy is based on substrate-assisted antibody catalysis and utilizes analogs designed to mimic the transition-state for intramolecular catalysis and thereby favor antibodies that can recruit catalytic groups from substrate. The hydrolysis of the benzoyl ester of cocaine provides an illustration. The benzoyl ester of cocaine is distant from the protonated nitrogen in the stable chair conformer but proximate in the strained boat form. An antibody stabilizing the boat form and approximating ester and amine could catalyze ester hydrolysis. To mimic the transition-state for the intramolecular catalysis, we synthesized a cocaine analog that replaces this ester with a methylenephenylphosphinate bridge to the tropane nitrogen. This bridged analog elicited 85 cocaine esterases out of 450 anti-analog antibodies-a performance markedly superior to that of a simple phosphonate ester-based analog with an identical tether. The correspondence of the analog to a "high energy" conformer eliminated product inhibition. For certain polyfunctional targets, substrate assistance can be an effective strategy for eliciting catalytic antibodies. PMID- 14747855 TI - Metallation of pyridines and quinolines in the presence of a remote carboxylate group. New syntheses of heterocyclic quinones. AB - 2-(3- and 2-Pyridylcarbonyl)benzoic acids (2,3), 2-(2-pyridylcarbonyl)thiophene-3 carboxylic acid (6), 2-(3-quinolylcarbonyl)benzoic acid (10), and most of the corresponding esters (compounds 1,7 and 9 ) are readily synthesized and involved in a deprotonation-condensation sequence. Biologically active aza-anthraquinones such as benzo[g]isoquinoline-5,10-dione (2-azaanthraquinone, 4 ) and benzo[g]quinoline-5,10-dione (1-azaanthraquinone, 5) are prepared using the strategy. Extension to other heterocyclic quinones such as thieno[3,2-g]quinoline 4,9-dione (8) and benzo[j]phenanthridine-7,12-dione (11) is also investigated. PMID- 14747856 TI - Directed, selective insertion of single molecules into patterned self-assembled monolayers of alkanethiols with different chain lengths. AB - Pd(ii) pincer adsorbate molecules (1) were inserted into self-assembled monolayers (SAMs) of alkanethiols with different chain lengths (C(8) to C(18)) on annealed gold substrates. Their presence was brought to expression by reaction of with Au nanoclusters bearing phosphine moieties (2). The surface-confined Au nanoclusters were observed only on the shorter chain SAMs (C(8)SH to C(16)SH) and not on C(18)SH SAMs. This is attributed to the longer chain length of C(18)SH preventing the insertion of pincer molecules. Microcontact printing (microCP) with C(18)SH on unannealed gold substrates and the subsequent immersion of the substrates into C(8)SH, C(10)SH, C(12)SH, or C(16)SH solutions, yielded a series of patterned SAMs that have areas of thiols of different chain lengths. Insertion of 1 followed by expression using 2, or insertion of 3 showed inserted molecules only in the shorter chain SAM areas. The absolute particle densities in the former case were higher than on the corresponding homogeneous SAMs on annealed substrates, probably due to larger numbers of defects in the SAMs on unannealed substrates. PMID- 14747857 TI - Phosphorus-nitrogen-phosphorus ligands: cooperative effects between nitrogen and phosphorus substituents on catalytic activity. AB - A new generation of PNP compounds bearing different diarylphosphine groups were prepared and used as ligands in palladium-catalysed Suzuki cross-coupling reactions. Rates of oxidative addition of iodobenzene to (PNP)Pd[0] complexes were measured using UV spectroscopy. Synergistic effects between the N- and P- substituents were identified and correlated in redox and catalytic chemistry. PMID- 14747858 TI - High affinity DNAzyme-based ligands for transition metal cations - a prototype sensor for Hg2+. AB - Inspired by recent interest in DNAzymes as transition metal ion sensors, a survey of the effects of various transition metals on the intramolecular cleavage rate of an imidazole modified, M(2+)-independent, self-cleaving "9(25)-11" DNA is reported. In particular, 9(25)-11 activity was strongly inhibited by Hg(2+)(K(d)(APP)= 110 +/- 9 nM). It is postulated that the affinity and selectivity of 9(25)-11 for Hg(2+) stems from the fact that this synthetically modified DNAzyme contains imidazoles. This study demonstrates the utility of modified nucleotides in developing DNAzyme sensors for metals ions, especially those for which unmodified nucleic acids might not serve as inherently good ligands. PMID- 14747859 TI - Effects of an 8-bromodeoxyguanosine incorporation on the parallel quadruplex structure [d(TGGGT)]4. AB - NMR, molecular dynamics and mechanics calculations, and CD spectroscopy were used to characterise three tetramolecular quadruplex complexes: [d(TG(Br)GGT)](4), [d(TGG(Br)GT)](4) and [d(TGGG(Br)T)](4), where G(Br) indicates an 8-bromoguanine residue. All three quadruplexes are characterised by a 4-fold symmetry with all strands parallel to each other and, differently to what has been observed for other parallel quadruplex structures, with a tetrad (formed by 8-Br-dGs) in a syn conformation. The whole of the data demonstrates that the replacement in turn of different dG residues with 8-Br-dG in the sequence 5[prime or minute]-TGGGT 3[prime or minute] affects the resulting structures in different ways, leading to different CD profiles and thermal stabilities. Particularly, [d(TG(Br)GGT)](4) and [d(TGG(Br)GT)](4) are more stable than the unmodified sequence, whereas [d(TGGG(Br)T)](4) is much less stable than the natural counterpart. The conformational features found in the three quadruplexes might, in principle, amplify the range of applicability of synthetic oligonucleotides as aptamers or catalysts, by providing novel structural motifs with different molecular recognition capabilities from those of native DNA sequences. PMID- 14747860 TI - Design and synthesis of multi-component 18 pi annulenic fluorofullerene ensembles suitable for donor-acceptor applications. AB - A series of trannulene (all-trans annulene) derivatives of [60]fullerene have been prepared by reacting C(60)F(18) with methanetricarboxylate esters that incorporate a range of photoactive functions. All the compounds have the intense emerald-green colour of fullerene trannulenes, characterised by strong bands at ca. 612 and 667 nm. Single crystal X-ray studies show that the packing varies with the nature of the addend, attributable to differing steric effects. UV/vis absorption spectra display transitions of the respective fullerene and addend models, indicating absence of electronic interactions between them in the ground state. These now provide an extensive series for testing photoactive (light harvesting) properties, with the exceptional properties of having strong visible light absorption. Their exceptional stability is attributed to the 18[small pi] aromatic circuit, inability to undergo nucleophilic substitution without disrupting this circuit, and a curved cage region that is shielded to reagents by the three bulky addends. PMID- 14747861 TI - Synthesis of rac-(1R,4aR,9aR)-2-methyl-1,3,4,9a-tetrahydro-2H-1,4a propanobenzofuro[2,3-c]pyridin-6-ol. An unusual double rearrangement leading to the ortho- and para-f oxide-bridged phenylmorphan isomers. AB - In an attempt to obtain the para-f isomer, rac-(1R,4aR,9aR)-2-methyl-1,3,4,9a tetrahydro-2H-1,4a-propanobenzofuro[2,3-c]pyridin-6-ol, via mesylation of an intermediate 9[small alpha]-hydroxyphenylmorphan, we obtained, instead, a rearranged chloro compound with a 5-membered nitrogen ring, 7-chloro-3a-(2,5 dimethoxyphenyl)-1-methyl-octahydroindole. This indole underwent a second rearrangement to give us the desired para-f isomer. The structures of the intermediate indole and the final product were unequivocally established by X-ray crystallography. A resynthesis of the known rac-(1R,4aR,9aR)-2-methyl-1,3,4,9a tetrahydro-2H-1,4a-propanobenzofuro[2,3-c]pyridin-8-ol, the ortho-f isomer, was achieved using the reaction conditions for the para-f isomer, as well as under Mitsunobu reaction conditions where, unusually, the oxide-bridge ring in the 5 phenylmorphan was closed to obtain the desired product. The synthesis of the para f isomer adds an additional compound to those oxide-bridged phenylmorphans that were initially visualized and synthesized; the establishment of the structure and configuration of 8 of the theoretically possible 12 racemates has now been achieved. The X-ray crystallographic structure analysis of the para-f isomer provides essential data that will be needed to establish the configuration of a ligand necessary to interact with an opioid receptor. PMID- 14747862 TI - Cyclodextrin complexation of a stilbene and the self-assembly of a simple molecular device. AB - (E)-4-tert-Butyl-4'-oxystilbene, 1(-), is thermally stable as the (E)-1(-) isomer but may be photoisomerized to the (Z)-1(-) isomer as shown by UV-vis and (1)H NMR studies in aqueous solution. When (E)-1(-) is complexed by alphaCD two inclusion isomers (includomers) form in which alphaCD assumes either of the two possible orientations about the axis of (E)-1(-) in alphaCD.(E)-1(-) for which (1)H NMR studies yield the parameters: k(1)(298 K)= 12.3 +/- 0.6 s(-1), DeltaH(1)(++)= 94.3 +/- 4.7 kJ mol(-1), DeltaS1(++)= 92.0 +/- 5.0 J K(-1) mol(-1), and k(2)(298 K)= 10.7 +/- 0.5 s(-1), DeltaH(2)(++)= 93.1 +/- 4.7 kJ mol(-1), DeltaS2(++)= 87.3 +/- 5.0 J K(-1) mol(-1) for the minor and major includomers, respectively. The betaCD.(E)-1(-) complex either forms a single includomer or its includomers interchange at the fast exchange limit of the (1)H NMR timescale. Complexation of 1(-) by N-(6(A)-deoxy- alpha-cyclodextrin-6(A)-yl)-N'-(6(A)-deoxy- beta cyclodextrin-6(A)-yl)urea, results in the binary complexes 2.(E)-1(-) in which both CD component annuli are occupied by (E)-1(-) and which exists exclusively in darkness and 2.(Z)-1(-) in which only one CD component is occupied by (Z)-1(-) and exists exclusively in daylight at lambda > or = 300 nm. Irradiation of solutions of the binary complexes at 300 and 355 nm results in photostationary states dominated by 2.(E)-1(-) and 2.(Z)-1(-), respectively. In the presence of 4 methylbenzoate, 4(-), 2.(Z)-1(-) forms the ternary complex 2.(Z)-1(-).4(-) where 4(-) occupies the second CD annulus. Interconversion occurs between 2.(Z)-1(-).4( ) and 2.(E)-1(-)+4(-) under the same conditions as for the binary complexes alone. Similar interactions occur in the presence of 4-methylphenolate and 4 methylphenylsulfonate. The two isomers of each of these systems represent different states of a molecular device, as do the analogous binary complexes of N,N-bis(6(A)-deoxy- beta-cyclodextrin-6(A)-yl)urea, 3, [3.(E)-1(-) and 3.(Z)-1( ), where the latter also forms a ternary complex with 4(-). PMID- 14747863 TI - Prodrugs of HIV protease inhibitors-saquinavir, indinavir and nelfinavir-derived from diglycerides or amino acids: synthesis, stability and anti-HIV activity. AB - With the aim of improving the pharmacological properties of current protease inhibitors (PIs), the synthesis of various acyl and carbamate amino acid- or diglyceride-containing prodrugs derived from saquinavir, indinavir and nelfinavir, their in vitro stability with respect to hydrolysis and their anti HIV activity in CEM-SS and MT4 cells have been investigated. l-Leucine (Leu) and l-phenylalanine (Phe) were connected through their carboxyl to the PIs while l tyrosine (Tyr) was conjugated through its aromatic hydroxyl via various spacer units. Hydrolysis of the prodrug with liberation of the active free drug was crucial for antiviral activity. The Leu- and Phe-PI prodrugs released the active free drug very rapidly (half-lives of hydrolysis in buffer at 37 degree C of 3-4 h). The Tyr-PI conjugates with a -C(O)(CH(2))(4)- linker exhibited half-lives in the 40-70 h range and antiviral activities in the 21-325 nM range (from 2 to 22 nM for the free PIs). The chemically very stable carbamate "peptidomimetic" Tyr PI prodrugs (no hydrolysis detected after 7 days in buffer) displayed a very low anti-HIV activity or were even inactive (EC(50) from 2300 nM to >10 microM). A very low antiviral activity was measured for the diglyceride-substituted saquinavir and for all of the disubstituted indinavir and nelfinavir prodrugs. All these prodrugs probably released the active parent PI too slowly under the antiviral assay conditions. These results combined with those from transepithelial transport studies (Rouquayrol et al., Pharm. Res., 2002, 19, 1704 1712) indicate that conjugation of amino acids (through their carboxyl) to the PIs constitutes a most appealing alternative which could improve the intestinal absorption of the PIs and reduce their recognition by efflux carriers. PMID- 14747864 TI - Evidence for gas-phase redox chemistry inducing novel fragmentation in a complex natural product. AB - The fragmentation of monensin A, in the presence of calcium, barium, silver and copper salts was studied by electrospray ionisation tandem accurate-mass mass spectrometry. The results showed that the calcium, barium and silver complexes of monensin A showed no significant alteration in their fragmentation to that previously observed for the sodium salts. However, the fragmentation of the copper(ii) salt resulted in new fragmentation routes. We propose that the copper might be initiating a novel gas-phase redox reaction resulting in a series of highly diagnostic ions. This methodology is demonstrated by locating the change in structure between the naturally occurring analogues monensin A and B. PMID- 14747865 TI - Asymmetric synthesis of the stereoisomers of 2-amino-5-carboxymethyl-cyclopentane 1-carboxylate. AB - The stereoisomers of 2-amino-5-carboxymethyl-cyclopentane-1-carboxylate may be prepared stereoselectively from diester derivatives of (E,E)-octa-2,6-diendioc acid, with the key step utilising the conjugate addition of homochiral lithium N benzyl-N- alpha-methylbenzylamide. The trans-C(1)-C(2)-stereoisomers are readily prepared via a diastereoselective tandem conjugate addition cyclisation protocol with lithium (R)-N-benzyl-N- alpha-methylbenzylamide, with subsequent hydrogenolysis and ester hydrolysis giving the (1R,2R,5R)- and (1R,2R,5S)- beta amino diacids in good yields. The preparation of the cis-C(1)-C(2)-stereoisomers utilises a protocol involving N-oxidation and Cope elimination of the major diastereoisomeric product arising from conjugate addition and cyclisation, giving homochiral (R)-5-carboxymethyl-cyclopentene-1-carboxylate. Conjugate addition of either lithium (R)- or (S)-N-benzyl-N- alpha-methylbenzylamide to (R)-5 carboxymethyl-cyclopentene-1-carboxylate, and diastereoselective protonation with 2,6-di-tert-butyl phenol gives, after hydrogenolysis and ester hydrolysis, the (1S,2R,5R)- and (1R,2S,5R)- beta-amino diacids in good yield. The use of (S)-N benzyl-N- alpha-methylbenzylamide in the initial conjugate addition and cyclisation reaction, and subsequent repetition of the elimination and conjugate addition strategy allows stereoselective access to all stereoisomers of 2-amino-5 carboxymethyl-cyclopentane-1-carboxylate. PMID- 14747866 TI - A two-directional synthesis of the C58-C71 fragment of palytoxin. AB - A two directional approach, in which asymmetric dihydroxylation and reduction reactions were used to control absolute configuration, was exploited in the preparation of a C(2)-symmetrical dipyranone. The homotopic dihydropyran (DHP) rings of this precursor were differentiated statistically using by a Prevost reaction and further functionalisation. A second Prevost reaction was used to functionalise the other DHP; global deprotection and peracetylation gave a protected version of the C(58)-C(71) fragment of palytoxin. Methods which might be of value in future synthetic work were developed for the stereoselective functionalisation of THP rings similar to those found in this fragment. PMID- 14747867 TI - Factors influencing solvent adduct formation by calixarenes in the solid state. AB - Structural studies of seven very differently functionalised derivatives of calix[4]arene have been used to provide an analysis of the numerous factors which may influence solvent adduct formation by calixarenes. Evidence is presented that even where a solvent guest is included within the calixarene cavity, interactions solely within the cavity cannot be seen as the sole influences upon the guest position and orientation. PMID- 14747868 TI - Selective esterifications of alcohols and phenols through carbodiimide couplings. AB - Esterification of carboxylic acids capable of forming ketene intermediates upon treatment with carbodiimides permits the selective acylation of alcohols in the presence of phenols lacking strong electron-withdrawing groups. The selectivity of acylations involving highly acidic phenols could be reversed through the addition of catalytic amount of acid. Esterification of other carboxylic acids was found to proceed through the formation of symmetric anhydrides and provide the opposite chemoselectivity. In both cases the relative acylation rates of substituted phenols are consistent with a reaction mechanism involving an attack of phenolate anions on electrophilic intermediates such as ketenes and symmetric anhydrides, with the carbodiimides serving both as an activating reagent and as a basic catalyst. PMID- 14747869 TI - Preparation of 6-O-(4-alkoxytrityl)celluloses and their properties. AB - Cellulose was reacted with a series of 4-alkoxytrityl chlorides (C(n)TCl, n: number of carbon atoms in a saturated alkyl chain) under homogeneous reaction conditions in LiCl-N,N-dimethyl acetoamide to give a series of 6-O-(4 alkoxytrityl)celluloses (C(n)TC) with a high degree of substitution (DS), from 0.94 to 0.99, and with high regioselectivity at the 6-O position. Solubility of the C(n)TC in nonpolar solvents depended on the alkyl chain length: as the alkyl chain lengthens, cellulose derivatives become more hydrophobic and are readily soluble in nonpolar solvents, but not in polar solvents. Acetates of the C(4) C(18)TC (C(4)-C(18)TCAc) showed anisotropic structures over melting temperatures (T(m)) examined under a polarized optical microscope (POM). Over isotropization temperatures (T(i)), flow birefringence were detected for C(12)-C(18)TCAc. The T(m) and T(i) decreased linearly with an increasing number of carbon atoms in the alkyl substituent. Wide-angle X-ray scattering (WAXS) studies of C(n)TC indicated that the fully extended side chains were perpendicular to the polymer backbone and interdigitated. These C(n)TC with the improved solubility may be used as starting materials for further derivatization focused on the secondary hydroxyl groups at the C-2 and C-3 positions. PMID- 14747870 TI - Enantioselective hydrolysis of various substituted styrene oxides with Aspergillus Niger CGMCC 0496. AB - Enantioselective biohydrolysis of various substituted styrene oxides using whole fungus cells of Aspergillus nigerCGMCC 0496 are described. The results show not only para- but also some ortho- substituted styrene oxides can achieve high enantioselectivity during the hydrolysis. PMID- 14747871 TI - Thermal decomposition of O-benzyl ketoximes; role of reverse radical disproportionation. AB - Thermolyses of seven dialkyl, two alkyl-aryl and two diaryl O-benzyl ketoxime ethers, R(1)R(2)C[double bond, length as m-dash]NOCH(2)Ph, have been examined in three hydrogen donor solvents: tetralin, 9,10-dihydrophenanthrene, and 9,10 dihydroanthracene. All the oxime ethers gave the products expected from homolytic scission of both the O-C bond (viz., R(1)R(2)C[double bond, length as m-dash]NOH and PhCH(3)) and N-O bond (viz., R(1)R(2)C[double bond, length as m-dash]NH and PhCH(2)OH). The yields of these products depended on which solvent was used and the rates of decomposition of the O-benzyl oxime ethers were greater in 9,10 dihydrophenanthrene and 9,10-dihydroanthracene than in tetralin. These results indicated that a reverse radical disproportionation reaction in which a hydrogen atom was transferred from the solvent to the oxime ether, followed by [small beta]-scission of the resultant aminoalkyl radical, must be important in the latter two solvents. Benzaldehyde was found to be an additional product from thermolyses conducted in tetralin. This, and other evidence, indicated that another induced decomposition mode involving abstraction of a benzylic hydrogen atom, followed by [small beta]-scission of the resulting benzyl radical, became important for some substrates. Participation by minor amounts of enamine tautomers of the oxime ethers was shown to be negligible by comparison of thermolysis data for the O-benzyloxime of bicyclo[3.3.1]nonan-9-one, which cannot give an enamine tautomer, with that of the O-benzyloxime of cyclohexanone. PMID- 14747872 TI - Preparation of beta-and gamma-lactams via ring closures of unsaturated carbamoyl radicals derived from 1-carbamoyl-1-methylcyclohexa-2,5-dienes. AB - 1-Carbamoyl-1-methylcyclohexa-2,5-dienes produced the corresponding delocalised 1 carbamoyl-1-methylcyclohexa-2,5-dienyl radicals on treatment with radical initiators. At temperatures above ca. 300 K dissociation to produce toluene and aminoacyl (carbamoyl) radicals took place. The alternative dissociation of the 1 carbamoyl-1-methylcyclohexa-2,5-dienyl radicals to release methyl radicals and an aromatic amide did not compete. Aminoacyl radicals with allyl, butenyl or similar side chains underwent cyclisations. Moderate yields of N-benzyl-azetidin-2-ones and N-benzyl-pyrrolidin-2-ones were isolated for a range of substituents. The main by-products were N-benzyl-N-alkenylformamides. Ring closure did not take place to a significant extent for precursors with alk-2-ynyl or 2-cyanoalkyl side chains. An improved yield of 1,3-dibenzylazetidin-2-one was obtained by use of lauroyl peroxide as initiator and by inclusion of methyl thioglycolate as polarity reversal catalyst. PMID- 14747873 TI - Pseudo-C3-symmetric trisoxazolines as ligands in copper catalyzed enantioselective Diels-Alder reaction. AB - Air- and water-stable chiral catalyst trisoxazolines4-6 /Cu(ClO(4))(2)[middle dot]6H(2)O have been used in the Diels-Alder reaction of cyclopentadiene with acryloyl-2-oxazolidinones or ketoesters. The reaction is carried out in air and up to 82% ee was achieved. PMID- 14747874 TI - Synthesis of various sulfoforms of the trisaccharide beta-d-GlcpA-(1-->3)- beta-d Galp-(1-->3)-beta-d-Galp-(1-->OMP) as probes for the study of the biosynthesis and sorting of proteoglycans. AB - A straightforward preparation of various sulfoforms of the trisaccharide 4 methoxyphenyl O-(sodium beta-d-glucopyranosyluronate)-(1-->3)-( beta-d galactopyranosyl)-(1-->3)-beta-d-galactopyranoside (1), namely its 6a- and 4a monosulfate, 6b- and 4b-monosulfate and 6a,6b-disulfate derivatives, is reported for the first time. These compounds, which are partial structures of the linkage region of proteoglycans, will serve as probes for the study of the biosynthesis and sorting of these macromolecules. A key trisaccharide derivative, in which the two similar d-Gal units were differentiated at C-4,6 with 4,6-benzylidene and 4,6 di-tert-butylsilylene acetals, respectively, was used as a common intermediate. Both acetal groups showed excellent orthogonality, and allowed the preparation of all target compounds in high yield. Noteworthy is the possibility to prepare the 6a- and 6b-monosulfated and the 6a,6b-disulfated species through a one-pot regioselective procedure starting from a tetrol precursor. PMID- 14747875 TI - Nontraumatic acute and subacute enhancing spinal epidural hematoma mimicking a tumor in a child. AB - We describe a 10-year-old boy who presented with acute onset of neck pain and neurologic symptoms caused by a spinal epidural hematoma. An enhancing mass, which mimicked a tumor, was seen on MR imaging of the cervical spine. The uptake of gadolinium in the mass and the associated mass effect suggested a tumor etiology. Such spinal epidural hematomas are extremely rare in the pediatric population. Additionally, it is even more uncommon for spinal epidural hematomas to resemble tumors. It is important to consider this entity when an enhancing epidural mass is found on MRI in the setting of an acute presentation. PMID- 14747876 TI - Anterior spinal cord infarction owing to possible fibrocartilaginous embolism. AB - Anterior spinal artery syndrome is characterised by acute flaccid quadriparesis or paraparesis, disturbance of pain and temperature sensation, and loss of sphincter control. Fibrocartilaginous embolism is a rarely recognised, but important cause of spinal cord infarction. Fibrocartilaginous embolisation usually occurs after minor trauma without major bony lesions, typically with an intervening symptom-free interval and progressive 'stroke-in-evolution' course. There is evidence that the embolus originates from the intervertebral disc, but the mechanism whereby disc fragments enter the spinal vessels is not well understood. We describe the evolution of MRI findings in a case of anterior spinal artery territory infarction thought to be secondary to fibrocartilaginous embolism. PMID- 14747877 TI - Fluid-fluid levels in transcalvarial Langerhans' cell histiocytosis. PMID- 14747879 TI - Diffusion-weighted MRI in herpes simplex encephalitis: a report of three cases. AB - Herpes simplex virus encephalitis (HSVE) is the most frequent fatal viral infection of the brain. Because antiviral treatment may improve the prognosis significantly, early diagnosis is mandatory. Imaging diagnosis rests on conventional MRI for the visualization of lesions in the limbic system, the hallmark of HSVE. Diffusion-weighted MRI (DW-MRI) has not been used for the evaluation of HSVE. We report on the DW-MRI findings in three patients with HSVE, who had cortical diffusion abnormalities in affected brain parenchyma, partially as the initial or most sensitive sign of encephalitis. Sequential imaging showed that the diffusion abnormality started to return to normal after 2 weeks in the presence of persistent contrast uptake. Thus, DW-MRI may be a valuable tool for early detection and diagnosis of HSVE whereas contrast-enhanced images are indispensable after the first week. PMID- 14747880 TI - The dural tail of intracranial meningiomas on fluid-attenuated inversion-recovery images. AB - We evaluated the "dural tail" associated with 48 intracranial meningiomas on fluid-attenuated inversion-recovery (FLAIR) and contrast-enhanced T1-weighted images. In 30 (62.5%), a dural tail was observed on contrast-enhanced T1-weighted images, and thickening of the dura mater and abnormal signal were identified in the corresponding region on FLAIR images. Thus, FLAIR imaging was useful for showing dural abnormality associated with meningiomas without the needed for contrast medium. PMID- 14747881 TI - Effects of valsartan compared with enalapril on blood pressure and cognitive function in elderly patients with essential hypertension. AB - OBJECTIVE: This prospective, randomised, open-label, blinded-endpoint study was to compare the effects of the angiotensin II (Ang II) AT1 receptor antagonist valsartan with those of the ACE inhibitor enalapril on blood pressure (BP) and cognitive functions in elderly hypertensive patients. METHODS: One hundred and forty-four patients aged 61-80 years with mild to moderate essential hypertension (DBP > or =95 mmHg and < or =110 mmHg at the end of a 2-week placebo run-in period) were randomly assigned to once daily (o.d.) treatment with valsartan 160 mg ( n=73) or enalapril 20 mg ( n=71) for 16 weeks. The patients were examined every 4 weeks during the study, with pre-dose BP (standard mercury sphygmomanometer, Korotkoff I and V) and heart rate (pulse palpation) being recorded at each visit. Cognitive function was evaluated at the end of the wash out period and after 16 weeks of active treatment by means of five tests (verbal fluency, the Boston naming test, word list memory, word list recall and word list recognition). RESULTS: Both valsartan and enalapril had a clear antihypertensive effect, but the former led to a slightly greater reduction in SBP/DBP at 16 weeks (18.6+/-4.6/13.7+/-4.0 mmHg vs 15.6+/-5.1/10.9+/-3.9 mmHg; P<0.01). Enalapril did not induce any significant changes in any of the cognitive function test scores; valsartan significantly increased the word list memory score (+11.8%; P<0.05 vs baseline and P<0.01 vs enalapril) and the word list recall score (+18.7%; P<0.05 vs baseline and P<0.01 vs enalapril), but not those of the other tests. CONCLUSION: These findings indicate that, in elderly hypertensive patients, 16 weeks of treatment with valsartan 160 mg o.d. is more effective than enalapril 20 mg o.d. in reducing BP, and (unlike enalapril) improves some of the components of cognitive function, particularly episodic memory. PMID- 14747882 TI - Phenotyping of N-acetyltransferase type 2 by caffeine from uncontrolled dietary exposure. AB - BACKGROUND AND OBJECTIVE: The standard approach for phenotyping of the human arylamine N-acetyltransferase 2 (NAT2) uses urinary caffeine metabolite ratios after a caffeine test dose taken in after methylxanthine abstinence. We tested whether these standardization measures were still needed when a more sensitive quantification technique was used. METHODS: A new liquid chromatography/tandem mass spectrometry (LC-MS/MS) method for the quantification of the caffeine metabolites 5-acetylamino-6-formylamino-3-methyluracil (AFMU), 5-acetylamino-6 amino-3-methyluracil (AAMU), 1-methylxanthine (1X), and 1-methylurate (1U) was developed. Urine samples from 77 healthy volunteers collected before and 5-6 h after oral intake of 150-200 mg caffeine were analyzed. The lower limits of quantification were 0.1 microg/ml for caffeine, 1X, 1U, and AFMU, and 0.2 microg/ml for AAMU. RESULTS: The urinary NAT2 ratios (AFMU+AAMU) / (AFMU+AAMU+1X+1U) before and after caffeine intake correlated well in 65 volunteers (r(2)=0.827; P< 0.0001). In 12 participants (16%), metabolite concentrations in urine before caffeine intake were below the quantification limit. NAT2 genotyping, done in 41 volunteers for four SNPs, corroborated the phenotyping results. CONCLUSION: NAT2 activity can be determined from a spontaneous urine probe in most subjects by quantification of caffeine metabolites arising from non-standardized dietary caffeine exposure using LC MS/MS. This may facilitate the phenotyping procedure. PMID- 14747883 TI - Comparisons of psychotropic drug prescribing patterns in acute psychiatric wards across Europe. AB - OBJECTIVE: To compare prescribed daily doses (PDDs) of psychotropic drugs in several European centres. METHOD: A one-day census of psychotropic drug prescriptions to 613 patients in 39 acute psychiatric wards in ten countries. RESULTS: Patients in Spain were on most drugs; patients in Germany were on the fewest. Chlorpromazine equivalents in Denmark, England, Germany and Spain were at high levels as were diazepam equivalents in Belgium, Finland, The Netherlands and Norway. Newer anti-psychotics were used in the majority of centres, although older anti-psychotics were used commonly in three centres. CONCLUSION: The high doses of psychotropic drugs patients receive in some centres may be having little additional therapeutic effect and could increase their risk of side effects. The use of older anti-psychotics in some centres may be causing side effects that could be reduced by using newer anti-psychotics. PMID- 14747884 TI - The effect of muscle pain on elbow flexion and coactivation tasks. AB - The effects of muscle pain on movement can easily be observed in daily life routines. However, the influence of muscle pain on motor control strategies has not been fully clarified. In this human experimental study it was hypothesized that muscle pain affects the motor control of elbow flexion movements, in different combinations of range of motion and target size, by decreased agonistic muscle activity and increased antagonistic muscle activity with consequent implications on kinematic parameters. The effects of experimentally induced muscle pain on movement strategy for: (1) small and large range of motion (ROM) elbow flexion movements towards a wide target, (2) large ROM flexion movements towards a narrow and wide target, and (3) subsequent coactivation of agonistic and antagonistic muscles to elbow flexion were assessed. Muscle pain induced by injections of hypertonic saline (1 ml, 5.8%) in either m. biceps brachii or m. triceps brachii caused similar effects on the movements. For low accurate movements the initial (100 ms) integrated electromyographic (EMG) activity of m. biceps brachii was decreased during muscle pain. In contrast, integrated EMG of the entire m. biceps brachii burst was decreased by muscle pain only for small ROM at a low accuracy, which also showed decreased EMG activity of m. triceps brachii and m. brachioradialis, together with increased activity of m. trapezius. Finally, high accurate movements and post-movement coactivation were generally not modulated by muscle pain. In summary, the present study shows that acute muscle pain can perturb the motor control strategy, which might be highly important in occupational settings where such a change may need compensatory actions from other muscles and thereby eventually contribute to the development of musculoskeletal pain problems. PMID- 14747885 TI - Greater impairment of extension movements as compared to flexion movements in Parkinson's disease. AB - Research on isometric contractions in subjects with Parkinson's disease (PD) has shown that anti-parkinsonian medication results in a greater increase in extensor strength than flexor strength. This finding is consistent with the hypothesis that there is a greater impairment in neural activation of extensor muscles as compared to flexor muscles in subjects with PD. Such a hypothesis is physiologically feasible given the known differences in the neural control of flexor and extensor muscles. If the above hypothesis is true for both phasic and tonic muscle activation, then differences between performance of rapid single joint flexion and extension movements should exist in subjects with PD. Twelve subjects with PD, "off" and "on" medication, and 12 age-and sex-matched healthy control subjects performed rapid single-joint movements in flexion and extension over three distances. For neurologically healthy subjects, we did not identify any significant differences in either kinematic or EMG parameters between flexion and extension movements. In contrast, in the PD subjects extension movements were slower and associated with more agonist bursts when compared to flexion movements. The results are consistent with the hypothesis that there is a differential impairment of neural activation of extensor muscles of the arm as compared to flexor muscles in subjects with PD. PMID- 14747887 TI - Predictiveness of a visual distractor modulates saccadic responses to auditory targets. AB - We are faster to orient our eyes toward a visual target that also produces a sound. Conversely, the response to an auditory target is prolonged if a visual distractor is presented at a spatially incongruent position. Here, participants exhibited faster saccadic reaction times when an auditory target was more likely to be presented opposite to a visual distractor than when the stimuli only rarely occurred in spatial disparity. In contrast to experiments with visual targets and an auditory distractor, a spatially congruent visual distractor did not facilitate the response to an auditory target. We interpret the results in terms of an ocular inhibition process to suppress an automatic orienting response to the location of the visual distractor. This process is shown to be modulated by the predictability of target location. PMID- 14747890 TI - In-capillary micro solid-phase extraction and capillary electrophoresis separation of heterocyclic aromatic amines with nanospray mass spectrometric detection. AB - A miniaturised technique to analyse and detect heterocyclic aromatic amines (HAs) using micro solid-phase extraction ( microSPE) coupled on-line (in-capillary) to capillary electrophoresis (CE) separation with nanospray (nESI) mass spectrometry (MS) detection has been developed. HAs are mutagenic and carcinogenic compounds formed at low levels in protein-rich food during cooking. Due to the low concentrations of HAs and the high complexity of the matrix in which they exist, sensitive and selective analytical methods are required for quantification. microSPE was performed on a packed bed of C(18 )particles inside the CE capillary, which minimised the dead volume. The on-line coupling of SPE, CE and nESI-MS reduced the time for extraction and identification to less than half an hour, which will allow for screening of several samples per day. The new technique provides short analysis time, low sample and solvent consumption, and HAs in standard solutions were easily detected at 12-17 fmol injections, and in spiked urine samples at 750-810 fmol injections. PMID- 14747891 TI - Use of D-T-produced fast neutrons for in vivo body composition analysis: a reference method for nutritional assessment in the elderly. AB - Body composition has become the main outcome of many nutritional intervention studies including osteoporosis, malnutrition, obesity, AIDS, and aging. Traditional indirect body composition methods developed with healthy young adults do not apply to the elderly or diseased. Fast neutron activation (for N and P) and neutron inelastic scattering (for C and O) are used to assess in vivo elements characteristic of specific body compartments. Non-bone phosphorus for muscle is measured by the (31)P(n, alpha)(28)Al reaction, and nitrogen for protein via the (14)N(n,2n)(13)N fast neutron reaction. Inelastic neutron scattering is used to measure total body carbon and oxygen. Body fat is derived from carbon after correcting for contributions from protein, bone, and glycogen. Carbon-to-oxygen ratio (C/O) is used to measure the distribution of fat and lean tissue in the body and to monitor small changes of lean mass. A sealed, D-T neutron generator is used for the production of fast neutrons. Carbon and oxygen mass and their ratio are measured in vivo at a radiation exposure of less than 0.06 mSv. Gamma-ray spectra are collected using large BGO detectors and analyzed for the 4.43 MeV state of carbon and 6.13 MeV state of oxygen, simultaneously with the irradiation. P and N analysis by delayed fast neutron activation is performed by transferring the patient to a shielded room equipped with an array of NaI(Tl) detectors. A combination of measurements makes possible the assessment of the "quality" of fat-free mass. The neutron generator system is used to evaluate the efficacy of new treatments, to study mechanisms of lean tissue depletion with aging, and to investigate methods for preserving function and quality of life in the elderly. It is also used as a reference method for the validation of portable instruments of nutritional assessment. PMID- 14747893 TI - Simultaneous determination of bismuth and tellurium in steels by high power nitrogen microwave induced plasma atomic emission spectrometry coupled with the hydride generation technique. AB - An annular-shaped, high power nitrogen microwave induced plasma (N(2)-MIP) produced at atmospheric pressure by an Okamoto cavity, as a new excitation source for atomic emission spectrometry (AES), has been used for the simultaneous determination of bismuth and tellurium in steels with the hydride generation method. Under the optimized experimental conditions, the best attainable detection limits at the Bi I 195.389 nm and Te I 200.200 nm lines were 110 and 86 ng/ml for bismuth and tellurium, respectively. The linear dynamic ranges for bismuth and tellurium were 300 to 30,000 ng/ml. The presence of several diverse elements was found to cause a more or less depressing interference with the proposed technique. When bismuth and tellurium in steels were determined, a large amount of Fe(III) in the solution caused a severe depressing interference, while the presence of Fe(II) showed little or no significant interference. Of the several interference-releasing agents examined, l-ascorbic acid was found to be the most preferable to reduce Fe(III) to Fe(II) prior to hydride generation. The concentrations of bismuth and tellurium in steels were determined by the proposed technique. The results obtained by this method were in good agreement with their certified values. PMID- 14747894 TI - Quantification of bile acids directly from plasma by MALDI-TOF-MS. AB - Matrix-assisted laser desorption/ionization (MALDI) time-of-flight mass spectrometry (MS) has proved to be a useful method for the quantification of bile acids directly from plasma. Six cholic acid derivatives were selected for analysis: taurocholic acid, taurochenodeoxycholic acid, taurolithocholic acid, glycocholic acid, glycochenodeoxycholic acid, and glycolithocholic acid. Solid phase extraction (SPE) columns were used to preconcentrate and purify the plasma samples. Calibration curves averaged from 3 days were obtained for the bile acids, and then tested for their ability to accurately determine concentrations from one measurement. In summary, a simple, rapid method has been developed for the quantification of bile salts from plasma by MALDI-MS with SPE cleanup. PMID- 14747895 TI - Quantitative SIMS depth profiling of diffusion barrier gate-oxynitride structures in TFT-LCDs. AB - Gate oxynitride structures of TFT-LCDs were investigated by SIMS, and successful solutions are demonstrated to overcome difficulties arising due to the charging effects of the multilayer systems, the matrix effect of the method, and the small pattern sizes of the samples. Because of the excellent reproducibility achieved by applying exponential relative sensitivity functions for quantitative analysis, minor differences in the barrier gate-oxynitride composition deposited on molybdenum capped aluminium-neodymium metallisation electrodes were determined between the centre and the edge of the TFT-LCD substrates. No differences were found for molybdenum-tungsten metallisations. Furthermore, at the edge of the glass substrates, aluminium, neodymium, and molybdenum SIMS depth profiles show an exponential trend. With TEM micrographs an inhomogeneous thickness of the molybdenum capping is revealed as the source of this effect, which influences the electrical behaviour of the device. The production process was improved after these results and the aging behaviour of TFT-LCDs was investigated in order to explain the change in control voltage occurring during the lifetime of the displays. SIMS and TEM show an enrichment of neodymium at the interface to the molybdenum layer, confirming good diffusion protection of the molybdenum barrier during accelerated aging. The reason for the shift of the control voltage was finally located by semi-quantitative depth profiling of the sodium diffusion originating from the glass substrate. Molybdenum-tungsten was a much better buffer for the highly-mobile charge carriers than aluminium-neodymium. Best results were achieved with PVD silicon oxynitride as diffusion barrier and gate insulator deposited on aluminium-neodymium metallisation layers. PMID- 14747897 TI - Voltammetric behaviour and determination of moxifloxacin in pharmaceutical products and human plasma. AB - The oxidative behaviour of moxifloxacin was studied at a glassy carbon electrode in different buffer systems using cyclic, differential pulse, and Osteryoung square-wave voltammetry. The oxidation process was shown to be irreversible over the entire pH range studied (2.0-10.0) and was diffusion-controlled. The methods were performed in Britton-Robinson buffer and the corresponding calibration graphs were constructed and statistical data were evaluated. When the proposed methods were applied at pH 6.0 linearity was achieved from 4.4 x 10(-7) to 1.0 x 10(-5) mol L(-1). Applicability to tablets and human plasma analysis was illustrated. Furthermore, a high-performance liquid chromatographic method with diode-array detection was developed. A calibration graph was established from 4.0 x 10(-6) to 5.0 x 10(-5) mol L(-1) moxifloxacin. The described methods were successfully employed with high precision and accuracy for estimation of the total drug content of human plasma and for pharmaceutical dosage forms of moxifloxacin. PMID- 14747899 TI - A one-step immunochromatographic assay for detecting ginsenosides Rb1 and Rg1. AB - An immunochromatographic strip test has been developed for detecting ginsenosides Rb1 (G-Rb1) and Rg1 (G-Rg1). This qualitative assay system is useful as a rapid screening method for detecting G-Rb1 and G-Rg1 in plants and plant preparations. Our assay is a competitive immunoassay that uses anti-G-Rb1 and anti-G-Rg1 monoclonal antibodies (MAbs) and a detection reagent that contains colloidal gold particles coated with anti-G-Rb1 and anti-G-Rg1 MAbs. Detection limits are 2 microg mL(-1) for both G-Rb1 and G-Rg1. PMID- 14747900 TI - Solvent extraction of amino acids into a room temperature ionic liquid with dicyclohexano-18-crown-6. AB - Amino acids Trp, Gly, Ala, Leu are extracted efficiently from aqueous solution at pH 1.5-4.0 (Lys and Arg at pH 1.5-5.5) into the room temperature ionic liquid 1 butyl-3-methylimidazolium hexafluorophosphate (BmimPF(6)) with dicyclohexano-18 crown-6 (CE). The most hydrophilic amino acids such as Gly are extracted as efficiently as the less hydrophilic (92-96%). The influence of pH, amino acid and crown ether concentration, volume ratio of aqueous and organic phases, and presence of some cations on amino acid recovery were studied. The ratio of amino acid to crown ether in the extracted species is 1:1 for cationic Trp, Leu, Ala, and Gly and to 1:2 for dicationic Arg and Lys. This ionic liquid extraction system was used successfully for the recovery of amino acids from pharmaceutical samples and fermentation broth, and was followed by fluorimetric determination. PMID- 14747901 TI - Wavelet packet transform and artificial neural network applied to simultaneous kinetic multicomponent determination. AB - This paper presents a novel method, named wavelet packet transform based multilayer feedforward neural network with Levenberg-Marquardt and back propagation algorithm (WPTLMBP), developed for simultaneous kinetic determination of Cu(II), Fe(III), and Ni(II). Wavelet packet representations of signals provided a local time-frequency description, thus in the wavelet packet domain the quality of noise removal can be improved. The artificial neural network was applied for non-linear multivariate calibration. In this study, by optimization, wavelet packet function, decomposition level and number of hidden nodes for WPTLMBP method were selected as Db2, 2, and 4 respectively. A program PWPTLMBP was designed to perform simultaneous kinetic determination of Cu(II), Fe(III), and Ni(II). The relative standard error of prediction (RSEP) for all components with WPTLMBP, LM-BP-MLFN, and PLS methods were 6.39, 10.4, and 8.30%, respectively. Experimental results showed the proposed method to be successful and better than the others. PMID- 14747903 TI - Botulinum toxin as an effective treatment of clozapine-induced hypersalivation. AB - Hypersalivation is a common and frequently disabling side effect of atypical neuroleptics such as clozapine. Current treatment options of this adverse advent are limited by lack of efficacy or additional side effects. Botulinum toxin (BTX) injections into the parotid glands have been shown to be very effective in treating sialorrhea in the context of various neurological disorders, such as Parkinson's and motor neuron disease. Surprisingly, BTX treatment of drug-induced sialorrhea has not yet been described. We here report a patient with clozapine induced hypersalivation and a good response to BTX injections lasting for more than 12 weeks, resulting in a marked reduction of the hypersalivation and consequently of his social withdrawal. Our patient serves to alert clinicians to the frequent problem of drug-induced sialorrhea and suggests that BTX injections should be considered as an effective and safe treatment for hypersalivation in psychiatric patients treated with clozapine. PMID- 14747902 TI - MDMA "ecstasy" alters hyperactive and perseverative behaviors in dopamine transporter knockout mice. AB - RATIONALE: Mice lacking the gene for the dopamine transporter (DAT) show a unique behavioral phenotype characterized by locomotor hyperactivity and repetitive circling in a novel environment. The hyperactivity of DAT (-/-) mice can be attenuated by psychostimulants and by serotonin uptake inhibitors, suggesting an important role for serotonin in the attenuation of locomotor hyperactivity in these mice. OBJECTIVES: These studies characterized the effects of 3,4 methylenedioxy-N-methylamphetamine (MDMA), a serotonin releaser, on the amount and patterns of locomotor activity in DAT (+/+) and (-/-) mice. We compared the locomotor patterns produced by MDMA to those observed in DAT (-/-) mice, and examined whether MDMA altered the hyperactivity and perseverative locomotor patterns in DAT (-/-) mice. METHODS: The effects of MDMA (10, 30 mg/kg) on locomotor activity in DAT (+/+) mice were measured for 90 min in a video tracker system to determine the optimal dose for subsequent studies in DAT (+/+) and (-/ ) mice. The effects of 20 mg/kg MDMA on patterns of locomotor activity in DAT (+/+) and (-/-) mice were measured for 90 min. RESULTS: In DAT (+/+) mice, MDMA increased locomotor activity and induced repetitive straight movement patterns. In DAT (-/-) mice, however, MDMA (20 mg/kg) attenuated the characteristic locomotor hyperactivity seen in these mice. In contrast, MDMA potentiated the thigmotaxis and decreased entropy observed in the DAT (-/-) mice. CONCLUSIONS: The effects of MDMA observed here demonstrate that the different aspects of the abnormal locomotor behavior exhibited by DAT (-/-) mice can be independently manipulated by pharmacological treatments. PMID- 14747904 TI - Post-traumatic overload or acute syndrome of the os trigonum: a possible cause of posterior ankle impingement. AB - The purpose of this paper is to discuss the post-traumatic overload syndrome of the os trigonum as a possible cause of posterior ankle impingement and hindfoot pain. We have reviewed 19 athletes who were referred to our foot unit between 1995 and 2001 because of posterior ankle pain, and in whom a post-traumatic overload syndrome of os trigonum was diagnosed. All these patients were followed up over a period of 2 years. In 11 cases a chronic repetitive movements in forced plantar flexion was found. In the other eight cases the pain appeared to persist after a standard treatment of an ankle sprain in inversion plantar flexion. The diagnosis was based on clinical history, physical examination and X-rays that revealed a non-fused os trigonum. The confirmation of diagnosis was carried-out injecting local anaesthetic under fluoroscopic control. In all cases a corticosteroid injection as first line treatment was performed. In 6 cases a second injection was necessary to alleviate pain because incomplete recovery with the first injection. Three cases (16%) were recalcitrant to this treatment and in these three cases a surgical excision of the os trigonum was carried out. Our conclusion is that after some chronic athletic activity or an acute ankle sprain the os trigonum, if present, may undergo mechanical overload, remain undisrupted and become painful. Treatment by corticosteroid injection often resolves the problem. PMID- 14747906 TI - [Conventional and semi-open kyphoplasty]. AB - Kyphoplasty is a young method which was developed for the minimally invasive augmentation of osteoporotic vertebral fractures. In contrast to vertebroplasty, the kyphoplasty technique allows an age-dependent fracture reduction through the inflation of a special balloon in the fractured cancellous bone of the vertebral body. The cancellous bone of the fracture zone is compressed by the balloon, so that a cavity remains in the vertebral body after removing the balloon, which is filled with highly viscous augmentation material. The reduced risk of serious complications, for example epidural leakage of augmentation material, justifies progressively expanding the indications for this technique to traumatic fractures with involvement of the posterior vertebral wall and neoplastic vertebral collapse due to osteolytic metastasis. Besides the indications for the conventional percutaneous approaches, the microsurgical interlaminary approach allows the use of kyphoplasty in more complex fractures involving compression of the neural structures. Kyphoplasty induces swift pain relief and allows rapid mobilisation of patients due to the immediate stabilisation of the affected vertebral bodies. Apart from the operative intervention, the medical treatment of the primary disease and the rehabilitation of the individual patient should be optimised through an interdisciplinary approach. PMID- 14747905 TI - [Percutaneous cementing techniques of the spine -- chances and limits]. AB - Vertebroplasty and Kyphoplasty represent minimal-invasive techniques for cement augmentation of vertebral bodies. Both procedures are successfully used for pain relieving stabilizations of osteoporotic fractures or malignant processes. Advantages of kyphoplasty over vertebroplasty are to be seen in the possibility of deformity correction as well as in a decreased risk of cement extrusions which represent the most important potential for clinical complications. Long-term experiences with the effect of cementing are sparse. Thus it seems even more important, to judge indications and possibilities realistically. The decision whether and when to perform an augmentation is influenced by multiple factors. These include age of the patient, age of the fracture, degree of deformation and further degenerative changes of the spine. This article summarizes the present research and literature und is thought to provide guidelines for the aforementioned decision making processes. PMID- 14747907 TI - [Vertebroplasty in severe osteoporosis. Technique and experience with multi segment injection]. AB - In severe osteoporosis progressive collapse of multiple vertebrae is an unsolved problem. Medical treatment appears to be too slow to prevent the course. The evolving experience with vertebroplasty led us to treat these problems with more extensive cement injections. Of 362 patients who were treated with percutaneous cement injection over a 5-year period, 100 were injected at five and more levels (average 7.3, maximum 14). The surgical technique has been refined, allowing six levels to be injected monolaterally under local anesthesia. No more than six levels or 25-30 cc of cement should be injected per session. The outcome of the procedure is favorable in 84% of patients with a significant pain decrease (from 7.6 to 2.7 VAS). More impressive is the subjective report of the patients about better posture and increased force in their back, allowing them to become more active again. The radiological follow-up for 1 year shows a stable situation without further sintering of the reinforced vertebrae and maintained disc space in between these vertebrae. Vertebroplasty on multiple levels is efficient and can prevent further collapse. Due the risk of fat embolism the injections should be limited to six levels per session. PMID- 14747908 TI - [Balloon kyphoplasty of vertebral compression fractures with a new calcium phosphate cement]. AB - QUESTION: Can the same levels of pain reduction and increase in function be achieved in kyphoplasty procedures with Calcibon as with polymethylmethacrylate (PMMA) cement? PATIENTS AND METHODS: In a prospective, interdisciplinary single center study, 99 patients (173 vertebral fractures) were treated with kyphoplasty. Augmentation was performed with PMMA in 66 cases (127 vertebral bodies) and with Calcibon in 33 patients (46 vertebral bodies). Outcome data were obtained with a VAS spine score and by radiomorphometric evaluation of X-rays before and after treatment. RESULTS: Pain and function improved in 87% of the patients; an average of 16% of the lost vertebral height was regained. A 9% cement leakage rate was observed with PMMA and 10% with Calcibon. There was no significant difference in pain reduction and radiomorphometric evaluation between the two techniques. CONCLUSION: Kyphoplasty is a reliable, minimally invasive method to stabilize fractured vertebral bodies. Augmentation with Calcibon improves pain and function and enables the treated vertebral body to regain of height. PMID- 14747909 TI - [Complications of vertebroplasty]. AB - Percutaneous vertebroplasty was first introduced in 1984 by Galibert et al. for the treatment of hemangiomas in the spine. The current indications for vertebroplasty also include compression fractures due to osteoporosis as well as osteolytic metastases and spinal myeloma lesions. With the numbers of percutaneous vertebroplasty performed by orthopedic and trauma surgeons, neurosurgeons, and radiologists steadily increasing, complications have also risen. Over the last 3 years an increasing number of cases with varying complications, their genesis, and their management have been reported in the literature. Complications include asymptomatic cement leakage, cardiovascular effects, embolism with lethal outcome as well as severe neurological deficits. This article presents a review of the complications reported in the literature, strategies for preventing possible complications as well as current concepts in therapy management. Several of our cases with cement leakages are presented. PMID- 14747910 TI - [Stress levels in bones and bone cement in the thoracolumbar spine afer kyphoplasty. Finite element study]. AB - The study quantified the stress levels in treated and untreated vertebral bodies following kyphoplasty. Three-dimensional FE models of treated and untreated T11, T12, L1, and L2 vertebral bodies were evaluated to examine the stress levels within the bone and bone cement. A validated T12-L1 model was used to investigate the effect of kyphoplasty treatment on adjacent vertebral stresses and strains. Using the single vertebral models, bone cement modulus changes had minimal effect on the stresses in the bone or the cement. The presence of bone cement had a minimal effect on the stress magnitudes or distribution in the adjacent T12 vertebra. This study provides quantification of the stress levels in bone cement and bone in vertebral bodies treated with kyphoplasty under in vivo-like loading conditions. The presence of bone cement immediately following kyphoplasty has only a slight effect on the stress magnitudes or distributions in adjacent vertebrae. PMID- 14747911 TI - [Radiotherapy of osteoarthritis. Indication, technique and clinical results]. AB - Regarding orthopedic topics, radiotherapy of degenerative joint disorders is actually not well researched. So far, adequate long-term observations and reliable assessment of symptoms according to objective orthopedic criteria are still missing. From 1984 to 1994, 85 patients with symptomatic knee joint and hip joint osteoarthritis as well as omarthritis and rhizarthritis were treated. A total of 73 patients or 103 joints (due to bilateral symptoms) were documented in a long-term follow-up using orthopedic scores including objective criteria. Of the patients previously resistant to therapy, 63% responded to RT. Endoprosthetic surgery was necessary for only three patients. With regard to the endpoints "complete pain relief" or "major pain relief," only the parameter "symptom exists 2 years or more" indicated a significantly negative prognosis in multivariate analysis ( p<0.05). Radiotherapy of degenerative joint disorders is an effective alternative treatment for refractory osteoarthritis compared to conventional conservative treatment options. PMID- 14747912 TI - [Titanium modular revision prosthesis stem in revision hip prosthesis]. AB - In revision arthroplasty of the hip, femoral defects and fractures must be bridged by a long stem. The implant has to be adapted exactly to the diameter of the cortical tube and has to be placed distally beyond the pathologic area. The titanium modular revision prosthesis (MRP) stem consists of the titanium alloy Ti6Al7Nb and is offered in various diameters, lengths, and special devices, which allows individual and load-stable reconstruction. The aim of the current study is to scrutinize first clinical and radiological results of the titanium MRP revision stem in revision arthroplasty of the hip. We report the midterm results of 130 non-cemented revision arthroplasties of the hip using the titanium MRP revision stem with a mean follow-up of 2.9 years (minimum: 1 year, maximum: 8 years), including 56 men and 74 women. The indication for revision arthroplasty was aseptic loosening of the cup and stem in 94 cases, loosening of the stem in 17 cases, reimplantation in a Girdlestone situation in 11, and periprosthetic fractures in 8 cases. The Harris hip score improved from 50.9+/-24.6 preoperatively to 88.5+/-11.5 postoperatively. The score according to Merle d'Aubigne improved from 9.8+/-4.0 preoperatively to 15.2+/-2.8 postoperatively. We found three recurrent dislocations, two lesions of the sciatic nerve, three cases of persisting deep infection, two periprosthetic fractures, and one case of subsidence of the stem of 2 cm required revision of the MRP stem. Atrophy of the proximal femur was visible in all cases with a stem diameter >17 mm (39/130). Stable primary fixation was achieved in 129 of 130 cases. The titanium MRP revision has proved to be a very reliable implant in cementless revision arthroplasty of the hip. PMID- 14747913 TI - [Growth and differentiation factor 5 (GDF-5) composite improves the healing of necrosis of the femoral head in a sheep model. Analysis of an animal model]. AB - MATERIALS AND METHODS: After creation of necrosis in 23 sheep, the composites were implanted in half of the animals. The animals were sacrificed 6 and 12 weeks after implantation and the femora were harvested. The specimens were investigated by microradiography, computed tomography, and histologically. RESULTS: The GDF-5 composites were effective in the necrosis model. Osseous regeneration of the necrosis and the drill track were accelerated and enhanced by the composites. In treated animals the necrosis was nearly healed and the drill track was filled with bone after 12 weeks. In the control group the track was partially filled with fibrous tissue and necrotic lesions were still present. Specific side effects of the growth factor or the matrix were not documented. This was documented by histological scoring and CT investigation. DISCUSSION: The application of an absorbable GDF-5 composite in combination with a core decompression procedure may enhance the healing of devitalized bone defects and is a promising approach for further studies. PMID- 14747916 TI - Mapping of quantitative trait loci (QTLs) affecting autumn freezing resistance and phenology in Salix. AB - Quantitative trait locus (QTL) analysis was performed at different time points during cold-acclimation of a tetraploid F(2 ) Salix pedigree. The pedigree ( n=92) was derived from a cross between a frost-susceptible diploid female clone 'Jorunn' ( Salix viminalis) and a frost resistant hexaploid male clone 'SW901290' ( Salix dasyclados). Freezing resistance, height growth increment and number of new leaves were assessed at days 0, 12, 20, 24, 31 and 42 of a short day-low temperature (SD-LT) hardening regime, while the initiation of shoot tip abscission and shoot tip abscission were measured daily. Height increment, dry-to fresh weight ratio and number of new leaves were also measured in a replicated field trial. Freezing resistance was determined from electrolyte leakage of leaf tissues and from visual injuries on stem segments, after exposure to a predetermined freeze-thaw stress. Using a genetic map of the F(2) composed of 432 single-dose AFLP markers, a total of 19 genomic regions controlling freezing resistance (10) and phenological traits (9) before and during cold-acclimation (SD-LT) were identified. The magnitude of the phenotypic variation explained by each freezing resistance locus varied over acclimation time (0-45%), and there was no time point at which all the QTLs could be detected. The single QTL detected for non-acclimated freezing resistance did not reach significance at any time point during cold-acclimation, suggesting an independent genetic relationship between non-acclimated and acclimated resistance to freezing in Salix. Five of the loci associated with freezing resistance shared common intervals with loci controlling phenological traits. Of the 14 QTLs controlling autumn freezing resistance and/or phenological traits in the indoors experiment, six (43%) were associated with autumn phenology-related traits, i.e. height increment, dry-to-fresh weight ratio and number of new leaves, measured in the field. A major locus with multi-trait association in both indoor and outdoor experiments was detected. PMID- 14747914 TI - [Molecular characterization of tissue-engineered articular chondrocyte transplants based on resorbable polymer fleece]. AB - Three-dimensional arrangement and subsequent transplantation of chondrocytic cells in resorbable polymers has been shown to be a promising technique for the treatment of cartilaginous defects. Engineering of artificial cartilage tissue includes dedifferentiation of chondrocytes in monolayer culture, the use of biodegradable matrices and polymer scaffolds, and re-expression of chondrocytic marker genes in three-dimensional culture. The aim of this study was to characterize molecularly the phenotypic changes occurring with autologous cartilage tissue engineering. Human articular chondrocytes were isolated, cultured in medium containing human serum, and expanded up to passage 3. Chondrocytes were embedded in human fibrinogen and in polyglactin-polydioxanon fleeces and cultured three-dimensionally up to 4 weeks. Dedifferentiation of chondrocytes in monolayers and formation of cartilage tissue in vitro or after subcutaneous transplantation into nude mice was assessed by gene expression analysis of typical chondrocytic genes, histology, and immunohistochemistry. The expansion of chondrocytes with human serum resulted in the induction of type I and type III collagens, whereas cartilage-specific type II collagen, cartilage oligomeric matrix protein, cartilage link protein, and aggrecan were repressed and induced again after three-dimensional arrangement of chondrocytes in polyglactin-polydioxanon. Transplantation experiments documented the synthesis of proteoglycan and cartilage-specific type II collagen in vivo. Three-dimensional arrangement of human articular chondrocytes in resorbable polyglactin polydioxanon fleeces supports chondrogenic differentiation and the formation of a hyaline-like cartilaginous matrix in vitro and in vivo. PMID- 14747917 TI - Cloning and characterization of a gamma-3 hordein mRNA (cDNA) from Hordeum chilense (Roem. et Schult.). AB - Hordeum chilense is a wild relative of H. vulgare, cultivated barley, that has been successfully used in the synthesis of amphiploids by crossing with Triticum spp. These amphiploids-named generically x Tritordeum-have been tested under field conditions, and one of them, the hexaploid tritordeum obtained following chromosome doubling of the hybrid H. chilense x T. turgidum, shows traits of interest inherited from the barley parent. Of great interest is the allelic variation observed in the endosperm storage proteins and their influence on the breadmaking and malting quality of tritordeum. We report here two mRNA (cDNA) sequences for a gamma-3 hordein from two accession lines of H. chilense, H1 and H7, and their characterization by quantitative real time (QRT)-PCR in the developing endosperm. Sequences were obtained by rapid amplification of cDNA ends and "edge-to-edge" amplification of open reading frames from cDNA of H. chilense. Eight putative single nucleotide polymorphisms and one codon insertion were identified in the sequences of the H1 and H7 gamma-3 hordeins. The deduced amino acid sequences showed similar features to that of the gamma-3 hordein and gamma gliadins from barley and wheat, respectively. While the repetitive motif (PQQQPF) is similar to that of the gamma-3 hordein from H. vulgare, there are 19 motif repeats in H. vulgare, whereas H. chilense shows 15 tandem repeats. The transcription of the genes encoding for the gamma-3 hordein were monitored by QRT PCR: in both lines maximum transcription occurred 12 days after flowering. PMID- 14747918 TI - Similarities of omega gliadins from Triticum urartu to those encoded on chromosome 1A of hexaploid wheat and evidence for their post-translational processing. AB - The omega-gliadins encoded on chromosome 1 of the A genome were purified from Triticum aestivum L. (2n=6 x=42, AABBDD) cv. Butte86, nullisomic 1D-tetrasomic 1A of cv. Chinese Spring (CS N1DT1A), and the diploid T. urartu (2n=2 x=14, AA ). Reverse-phase high-performance liquid chromatography combined with sodium dodecyl sulfate-polyacrylamide gel electrophoresis of gliadin extracts from CS nullisomic tetrasomic (NT) lines confirmed the assignment to chromosome 1A. The purified omega-gliadins were characterized by mass spectrometry and N-terminal sequencing. The 1A-encoded omega-gliadins were smaller than 1B- or 1D-encoded omega-gliadins. The N-terminal amino acid sequences for 1A omega-gliadin mature peptides were nearly identical to those for the T. urartu omega-gliadins and were more similar to 1D omega-gliadin sequences than to sequences for T. monococum omega-gliadins, barley C-hordeins, or rye omega-secalins. They diverged greatly from the N terminal sequences for the 1B omega-gliadins. The data suggest that T. urartu is the A-genome donor, and that post-translational cleavage by an asparaginyl endoprotease produces those omega-gliadins with N-terminal sequences beginning with KEL. PMID- 14747920 TI - [The medical cooperative "Association of Urologists"]. PMID- 14747919 TI - rym15 from the Japanese cultivar Chikurin Ibaraki 1 is a new barley mild mosaic virus (BaMMV) resistance gene mapped on chromosome 6H. AB - Breeding for resistant cultivars is the only way to prevent high yield loss in barley caused by the soil-borne barley mild mosaic virus (BaMMV) complex. We have characterized the BaMMV resistance of barley cv. Chikurin Ibaraki 1. Doubled haploid lines were obtained from the F(1) between the susceptible six-rowed winter barley cultivar, Plaisant, and Chikurin Ibaraki 1. Each line was tested for reaction to BaMMV by mechanical inoculation followed by DAS-ELISA. Of 44 microsatellites that covered the genome, 22 polymorphic markers were tested on one susceptible and one resistant bulk, each comprising 30 lines. Differential markers and additional microsatellite markers in the same region were then tested on the whole population. A bootstrap analysis was used to compute confidence intervals of distances and to test the orders of the resistance gene and the closest markers. A segregation of 84 resistant/98 susceptible lines fitted a 1:1 ratio (chi(2)=1.08, P=0.30), which corresponds to a single gene in this DH lines population. The resistance gene was flanked by two markers near the centromeric region of chromosome 6HS-Bmag0173, at 0.6+/-1.2 cM, and EBmac0874, at 5.8 +/- 3.4 cM. We propose to name this new resistance gene rym15. This resistance gene and associated markers will increase the possibilities to breed efficiently for new cultivars resistant to the barley mosaic disease. PMID- 14747921 TI - [An innovative cooperative model between a department of urology and a urological medical specialist practice]. PMID- 14747922 TI - [Thoughts on regional networks of established urologists in the example of the Dusseldorf Regional Urologists' Network (UND)]. PMID- 14747923 TI - [Cooperative patient care at the Euromed Clinic]. PMID- 14747924 TI - [UCM Urology Centrum Munchen AG. The Munich model of a network of urologists]. PMID- 14747925 TI - [Fusion of urology practices. The Urological Practice Center Darmstadt]. PMID- 14747926 TI - [The radical retropubic prostatectomy -- gold standard for prostate cancer?]. AB - Radical retropubic prostatectomy is still the most frequent surgical procedure for the treatment of localized prostate cancer. Improvements in the surgical techniques have contributed to significant reduction in operative time (median 69 min) and in rates of early and late complications. The surgical procedure and perioperative management are described, and results in 180 consecutive cases are presented. Severe postoperative complications did not occur. More than 95% of patients were completely dry within 6 months. Erectile function recovered in 71% of patients with bilateral preservation of the neurovascular bundles and in 34% of patients with unilateral nerve sparing. These results are in good agreement with data from the literature and confirm the outstanding role of the retropubic approach as the gold standard for radical prostatectomy, as well as the reproducibility of the result in community-based hospitals. PMID- 14747927 TI - [Optimizing the use of radiotherapy with IMRT and image guided location of advanced prostate cancer]. AB - Intensity modulated radiotherapy (IMRT) combined with recently developed noninvasive image-guided targeting techniques for tumor localization/repositioning provide a means to further improve on conformal radiotherapy of prostate cancer by optimally sparing the rectum. This refined approach may potentially improve treatment results for locally advanced prostate cancer while reducing side effects. This review summarizes the clinical requirements for effective prostate radiotherapy and describes the new technology that helps to better fulfil these requirements. These noninvasive developments, their potential benefit as well as their limitations, together with new data on fractionation sensitivity of prostate cancer that may lead to shortened overall treatment times may be of interest for all physicians treating patients with prostate cancer. PMID- 14747929 TI - [Misdiagnosis of a Leiomyosarcoma of the inferior vena cava as a renal cell carcinoma]. AB - Leiomyosarcoma of the vena cava inferior (VCI) is a rare mesenchymal malignant tumor of the retroperitoneum. About 200 cases have been described in the literature so far. Leiomyosarcomas may resemble other tumors including renal cell carcinoma, adrenal carcinoma, and hepatic adenoma. Thus, misinterpretation of a leiomyosarcoma of the VCI is a common problem. We present a 71-year-old female with a large subhepatic, retroperitoneal tumorous mass which was diagnosed to be a renal cell carcinoma by sonography and computed tomography. After application of magnetic resonance imaging and retrospective analysis of the CT scan, diagnosis could be revised. Laparotomy and en bloc resection of the tumor was performed; histopathological examination confirmed a leiomyosarcoma of the VCI. The postoperative course was unremarkable. Leiomyosarcoma of the VCI may resemble advanced stages of renal cell carcinoma. High-resolution imaging modalities (computed tomography, magnetic resonance imaging) are able to precisely evaluate the typical imaging characteristics of leiomyosarcomas, thus determining correct diagnosis in affected patients. This is essential for successful operative therapy. PMID- 14747928 TI - [The impact of spinal cord injury on sexuality and reproduction]. AB - Spinal cord injury is known to have a major impact on human sexual function. The disturbances depend on the level and completeness of the lesion. The majority of people affected by spinal cord injury are young and in their reproductive years. In these cases sexuality reflects an important aspect of personality, and therefore special attention needs to be given to this issue during rehabilitation. Sexual rehabilitation encompasses both comprehensive information and consultation as well as diagnosis and treatment of disturbed sexual functions. Successful rehabilitation and integration depend also on restoration of sexual function or adequate therapy of sexual dysfunction. This review will consider the present knowledge about the impact of spinal cord injury on female and male sexual function, the currently available treatment options as well as the aspects of fertility and reproduction in this patient population. PMID- 14747930 TI - [The appraisal of physician liability. Conversation with Prof. K.-H. Bichler, Tubingen]. PMID- 14747931 TI - The industrial sustainability of bioremediation processes. PMID- 14747932 TI - Selective antimicrobial action of chitosan against spoilage yeasts in mixed culture fermentations. AB - The effect of chitosan on Saccharomyces cerevisiae (the yeast that carries out alcohol fermentation), Brettanomyces bruxellensis and Brettanomyces intermedius (contaminants of alcohol fermentations), was investigated. The effect of chitosan was tested on each yeast, as well as on mixed cultivations of S. cerevisiae + B. bruxellensis and S. cerevisiae + B. intermedius. Chitosan enhanced the lag period of both strains of Brettanomyces (80 h for B. bruxellensis and 170 h for B. intermedius with 6 and 2 g/l chitosan, respectively). The growth rate of S. cerevisiae was inversely proportional to the chitosan concentration; the former was 50% when 6 g/l polysaccharide was used. Moreover, in mixed cultivations of S. cerevisiae and Brettanomyces strains, it was found that both B. bruxellensis and B. intermedius failed to grow while growth of S. cerevisiae was not affected (using 3 and 6 g/l chitosan, respectively). An interesting collateral result was that the presence of chitosan accelerated the consumption of glucose in the mixed cultivations (60 h instead of 120 h). PMID- 14747933 TI - Medical Image Resource Center--making electronic teaching files from PACS. AB - A picture archive and communications system (PACS) is a rich source of images and data suitable for creating electronic teaching files (ETF). However, the potential for PACS to support nonclinical applications has not been fully realized: at present there is no mechanism for PACS to identify and store teaching files; neither is there a standardized method for sharing such teaching images. The Medical Image Resource Center (MIRC) is a new central image repository that defines standards for data exchange among different centers. We developed an ETF server that retrieves digital imaging and communication in medicine (DICOM) images from PACS, and enables users to create teaching files that conform to the new MIRC schema. We test-populated our ETF server with illustrative images from the clinical case load of the National Neuroscience Institute, Singapore. Together, PACS and MIRC have the potential to benefit radiology teaching and research. PMID- 14747934 TI - Security middleware infrastructure for DICOM images in health information systems. AB - In health care, it is mandatory to maintain the privacy and confidentiality of medical data. To achieve this, a fine-grained access control and an access log for accessing medical images are two important aspects that need to be considered in health care systems. Fine-grained access control provides access to medical data only to authorized persons based on priority, location, and content. A log captures each attempt to access medical data. This article describes an overall middleware infrastructure required for secure access to Digital Imaging and Communication in Medicine (DICOM) images, with an emphasis on access control and log maintenance. We introduce a hybrid access control model that combines the properties of two existing models. A trust relationship between hospitals is used to make the hybrid access control model scalable across hospitals. We also discuss events that have to be logged and where the log has to be maintained. A prototype of security middleware infrastructure is implemented. PMID- 14747935 TI - Effect of room illuminance on monitor black level luminance and monitor calibration. AB - In this article we demonstrate the effect of room illuminance and surrounding monitor black level luminance on image quality for soft copy interpretation. Luminance values of a 10% central target and image quality evaluations and observer performance using a contrast-detail mammography (CDMAM) phantom demonstrate these effects. Our results indicate that high room illuminance has a more damaging effect on image quality when the surrounding monitor luminance is 0% to 5% of the maximum monitor luminance. The effect of room illuminance is less obvious when the surrounding monitor luminance is 20% of the maximum. PMID- 14747936 TI - Evolution of the digital revolution: a radiologist perspective. AB - The transformation from film-based to filmless operation has become more and more challenging, as imaging studies expand in size and complexity. To adapt to these changes, radiologists must proactively develop new workflow strategies to compensate for increasing work demands and the existing workforce shortage. This article addresses the evolutionary changes underway in the radiology interpretation process and reviews changes that have occurred in the past decade. These include a number of developments in soft-copy interpretation, which is migrating from a relatively static process, duplicating film-based interpretation, to a dynamic process, using multi-planar reconstructions, volumetric navigation, and electronic decision support tools. The result is optimization of the human-computer interface with improved productivity, diagnostic confidence, and interpretation accuracy. PMID- 14747938 TI - Continuous venovenous haemofiltration in cancer patients with renal failure: a single-centre experience. AB - OBJECTIVE: To assess the effect of continuous venovenous hemodiafiltration (CVVHDF) in cancer patients with acute renal failure. PATIENTS AND METHODS: Retrospective study of all patients with acute renal failure requiring dialysis and treated with CVVHDF in a medical intensive care unit (ICU) from a cancer hospital. RESULTS: From January 1997 until December 2002, 32 cancer patients were treated with CVVHDF for acute renal failure. Their characteristics were: male/female 23/9, median age 61 years, haematological/solid tumours 16/16, and median APACHE II and IGS II scores 31/67. The number of organ failures was 1/2/3/4 in respectively 10/6/13/2 patients. Complete, partial or absence of resolution of acute renal failure was noted in 13, 8 and 11 patients. Sixteen patients (50%) died in the ICU and 15 (47%) were discharged alive from the hospital. In univariate analysis, variables statistically significantly adversely associated with hospital mortality were renal failure of renal origin, bone marrow transplant, increasing number of organ failures, reduced lymphocyte count, elevated bilirubin and lower creatinine levels, increased thromboplastin time, younger age, increased APACHE II and IGS II, ARDS and mechanical ventilation. In multivariate analysis, two models were used including either APACHE II or IGS II. The number of organ failures was found as the only significant prognostic factor in both models ( p=0.01). Elevated phosphate level was a poor prognostic factor for hospital mortality ( p=0.04) in the model including APACHE II. CONCLUSIONS: In the experience of a single centre, CVVHDF is effective in the treatment of acute renal failure in cancer patients. The increasing number of organ failures was the single independent poor predictive factor for hospital mortality. Cancer characteristics and general gravity scores were not predictive factors. PMID- 14747937 TI - Psychiatric morbidity among cancer patients and awareness of illness. AB - A significant proportion of cancer patients experience psychiatric morbidity. Potential predictors of psychiatric morbidity include patient disease-related factors and factors relating to the patient's environment. The aim of this study was to investigate the prevalence of psychiatric morbidity and the relationship between the clinical or personal factors, especially psychiatric morbidity, and awareness of cancer diagnosis among a group of Turkish cancer patients. A total of 117 cancer patients were assessed using the Structured Clinical Interview for DSM-IV (SCID), the Hospital and Anxiety Depression Scale (HADS) and the General Health Questionnaire (GHQ). Of these patients, 30% had a psychiatric diagnosis. Adjustment disorders comprised most of the psychiatric diagnoses. Awareness of the diagnosis of cancer, history of previous psychiatric disorders, pain and stress factors were correlated with psychiatric morbidity. Of the 117 patients, 64 (54.7%) were unaware of the diagnosis of cancer. Most of the patients (67.9%) who were considered to be aware of the cancer diagnosis stated that they had guessed their illness from the treatment process or drug adverse effects. Psychiatric morbidity was significantly higher in the patients who knew that they had a cancer diagnosis (P=0.03). These findings suggest that the awareness of cancer diagnosis is related to the presence of psychiatric morbidity. In particular, the understanding of the diagnosis indirectly may be stressful to the patient because it arouses suspicion about the cancer and treatment, and consequently can lead to psychiatric disturbance. In Turkey honest disclosure of the true diagnosis is still not common for cancer patients and it seems to be essential to improve this situation. PMID- 14747939 TI - Expression of nephrin in acquired forms of nephrotic syndrome in childhood. AB - Nephrin is a podocyte adhesion molecule located at the slit diaphragm between adjacent glomerular epithelial cells. Mutations in the gene encoding nephrin result in the absence of nephrin or alterations in nephrin causing massive proteinuria in patients with congenital nephrotic syndrome. Given the importance of nephrin to the structural integrity of the glomerular filtration barrier, we postulated that it might also be altered in acquired forms of nephrotic syndrome (NS). To test this hypothesis, frozen kidney biopsy sections from 29 pediatric patients with acquired NS and 5 controls were examined for expression of nephrin. The pathological diagnoses were minimal change disease (MCNS) (19) and focal segmental glomerulosclerosis (FSGS) (10). To determine if nephrin expression differed between children and adults with NS, 10 adult patients and 3 controls were also examined. Nephrin expression was evaluated by immunoperoxidase staining with a monoclonal antibody against the extracellular FnIII portion of human nephrin. In all cases, nephrin expression was seen along the glomerular basement membrane in a finely granular/linear pattern. Expression of nephrin was similar to controls in all 19 patients with MCNS and all 10 patients with FSGS. Areas of sclerosis in patients with FSGS did not demonstrate nephrin expression. A distinctly granular pattern to nephrin expression was seen in adult patients with NS as well as controls. These findings suggest that an alteration in nephrin expression is not a feature of acquired forms of NS in childhood. PMID- 14747940 TI - Potent cytotoxins produced by a microbial symbiont protect host larvae from predation. AB - Larvae of the sessile marine invertebrate Bugula neritina (Bryozoa) are protected by an effective chemical defense. From the larvae, we isolated three bryostatin class macrocyclic polyketides, including the novel bryostatin 20, that deterred feeding by a common planktivorous fish that co-occurs with B. neritina. A unique bacterial symbiont of B. neritina, Endobugula sertula, was hypothesized as the putative source of the bryostatins. We show that: (1) bryostatins are concentrated in B. neritina larvae and protect them against predation by fish; (2) the adults are not defended by bryostatins; and (3) E. sertula produces bryostatins. This study represents the first example from the marine environment of a microbial symbiont producing an anti-predator defense for its host and, in this case, specifically for the host's larval stage, which is exceptionally vulnerable to predators. PMID- 14747941 TI - Expression of galectin-3 in the rat pancreas during regeneration following hormone-induced pancreatitis. AB - Supramaximal dosage of the cholecystokinin analog caerulein leads to edematous pancreatitis with subsequent acinar cell destruction predominantly by apoptosis. We have used immunohistochemistry to reveal the expression of the anti-apoptotic protein galectin-3 in pancreatic acinar cells. Galectin-3, which occurs only in duct cells under physiological conditions, is expressed in a subset of acinar cells after the end of a 12-h caerulein infusion, giving rise to a "patchy" staining pattern. During the subsequent period of inflammation and regeneration, galectin-3 expression increases in those acinar cells that undergo apoptosis. By 48 h after the end of caerulein infusion, morphologically normal cells do not contain galectin-3 and participate in regeneration by proliferation. Tubular complexes, being transient structures from degenerative acini, accumulate galectin-3 in the remnants of the epithelium cells. Stimulation with supramaximal dosages of caerulein of the cell line AR4-2J, which is derived from rat pancreatic acinar cells, also results in a marked increase of galectin-3, confirming the in vivo results. We postulate that the high expression of the anti apoptotic protein galectin-3 regulates the time course of the apoptotic process in pancreatic acinar cells. PMID- 14747942 TI - Creation of EcR isoform-specific mutations in Drosophila melanogaster via local P element transposition, imprecise P element excision, and male recombination. AB - Collections of single P transposable-element insertion strains that currently inactivate more than 25% of essential Drosophila genes have proven to be a valuable tool for genome research in Drosophila melanogaster. For genes unrepresented in these collections, strategies including local P element transposition and transposase-induced imprecise excision can be used to inactivate or delete the gene of interest. Here we report our use of local P element transposition followed by imprecise P element excision and transposase induced male recombination to generate two deficiencies specific for the EcR-A isoform of the ecdysone receptor ( EcR) gene, and four larger deficiencies likely to affect multiple EcR functions. We also report here the determination of sequences flanking six EcR-B deficiencies generated in a previous imprecise excision screen. EcR-A encodes one of a family of three related nuclear receptor proteins that, together with the heterodimer partner USP, mediate ecdysone signaling during Drosophila development. Our results delineate sequences required in vivo for EcR-A function, as well as identifying EcR-A intron 1 sequences that are not essential for EcR function. PMID- 14747943 TI - Characterisation of a beta-tubulin gene from the monogenean parasite, Gyrodactylus salaris Malmberg, 1957. AB - The cDNA and two partial genomic sequences of beta-tubulin genes have been isolated from the monogenean parasite Gyrodactylus salaris. The cDNA sequence is not represented by either of the genomic sequences, implying that at least three isotypes of the gene exist in G. salaris. The sequences show regions of high homology with other helminth beta-tubulin genes. This represents the first isolation of a beta-tubulin gene from a monogenean and contributes to the overall characterisation of these genes within the helminths. This is an important area, as anthelmintic resistance is increasing against benzimidazole drugs that target the beta-tubulin gene. Benzimidazole drugs have been tested successfully against Gyrodactylus parasites, but their use is not widespread. Should it increase, analysis of the beta-tubulin gene may provide a tool for monitoring resistance development and improving management practises. Use of the beta-tubulin gene in the identification of Gyrodactylus species may prove complex due to the presence of different isotypes. PMID- 14747946 TI - A novel gene, the protein product of which is mainly expressed in germline cells and in the dorsal structures of Xenopus. AB - A novel cDNA was isolated from a Xenopus cDNA library using an antibody recognizing the germ plasm-specific germinal granules. The protein product of the gene ( Xdsg) was detected on the germinal granules of the cleaving embryo as determined by immunoelectron microscopy. The spatio-temporal distribution of the RNA transcript and protein product of the gene was investigated in oocytes and embryos. The protein was detected in the germ plasm and the germline cells from the cleavage to the tadpole stages, while both the protein and RNA transcript were prominent in the dorsal structures from the tailbud stage onward. The present study has shown for the first time that the protein product of the Xdsg gene was present on the germinal granules in Xenopus. PMID- 14747944 TI - Molecular genetic pathways in various types of endometrial carcinoma: from a phenotypical to a molecular-based classification. AB - Two types of endometrial carcinoma are distinguished with respect to biology and clinical course. Type-I carcinoma is related to hyperestrogenism by association with endometrial hyperplasia, frequent expression of estrogen and progesterone receptors and younger age, whereas type-II carcinoma is unrelated to estrogen, associated with atrophic endometrium, frequent lack of estrogen and progesterone receptors and older age. Histologically, endometrioid and mucinous carcinomas are considered type I, serous and clear cell carcinomas type II. Molecular data from multiple studies support the hypothesis of different genetic pathways in the development of endometrioid and serous carcinoma. The most frequent genetic alteration in endometrioid carcinoma is PTEN inactivation by mutation, followed by microsatellite instability (MIN) and mutations of K-ras and beta-catenin. PTEN and K-ras mutations and MIN are considered early events, occurring in a subset of atypical endometrial hyperplasia, whereas p53 mutation is considered a late event, during progression of about 10-20% of endometrioid carcinomas. In serous carcinoma, p53 mutation is the most frequent genetic alteration, followed by inactivation of p16 and e-cadherin and amplification of her2/neu. p53 mutation occurs in endometrial intraepithelial carcinoma, the putative precursor of serous carcinoma. Considering these genetic pathways, the current histological classification of endometrial carcinoma is molecular based. PMID- 14747947 TI - Recovery of photosystem I and II activities during re-hydration of lichen Hypogymnia physodes thalli. AB - Photochemical efficiencies of photosystem I (PSI) and photosystem II (PSII) were studied in dry thalli of the lichen Hypogymnia physodes and during their re hydration. In dry thalli, PSII reaction centers are photochemically inactive, as evidenced by the absence of variable chlorophyll (Chl) fluorescence, whereas the primary electron donor of PSI, P700, exhibits irreversible oxidation under continuous light. Upon application of multiple- and, particularly, single turnover pulses in dry lichen, P700 oxidation partially reversed, which indicated recombination between P700(+) and the reduced acceptor F(X) of PSI. Re-wetting of air-dried H. physodes initiated the gradual restoration of reversible light induced redox reactions in both PSII and PSI, but the recovery was faster in PSI. Two slow components of P700(+) reduction occurred after irradiation of partially and completely hydrated thalli with strong white light. In contrast, no slow component was found in the kinetics of re-oxidation of Q(A)(-), the reduced primary acceptor of PSII, after exposure of such thalli to white light. This finding indicated the inability of PSII in H. physodes to provide the reduction of the plastoquinone pool to significant levels. It is concluded that slow alternative electron transport routes may contribute to the energetics of photosynthesis to a larger extent in H. physodes than in higher plants. PMID- 14747948 TI - Rapid accumulation and metabolism of polyphosphoinositol and its possible role in phytoalexin biosynthesis in yeast elicitor-treated Cupressus lusitanica cell cultures. AB - Inositol 1,4,5-trisphosphate [Ins(1,4,5)P(3)] rapidly accumulates in elicited Cupressus lusitanica Mill. cultured cells by 4- to 5-fold over the control, and then it is metabolized. Correspondingly, phospholipase C (PLC) activity toward phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P(2)] is stimulated to high levels by the elicitor and then decreases whereas Ins(1,4,5)P(3) phosphatase activity declines at the beginning of elicitation and increases later. These observations indicate that elicitor-induced biosynthesis and dephosphorylation of Ins(1,4,5)P(3) occur simultaneously and that the Ins(1,4,5)P(3) level may be regulated by both PtdIns(4,5)P(2)-PLC and Ins(1,4,5)P(3) phosphatases. Studies on the properties of PLC and Ins(1,4,5)P(3) phosphatases indicate that PLC activity toward PtdIns(4,5)P(2) was optimal at a lower Ca(2+) concentration than activity toward phosphatidylinositol whereas Ins(1,4,5)P(3) phosphatase activity is inhibited by high Ca(2+) concentration. This suggests that Ins(1,4,5)P(3) biosynthesis and degradation may be regulated by free cytosolic Ca(2+). In addition, a relationship between Ins(1,4,5)P(3) signaling and accumulation of a phytoalexin (beta-thujaplicin) is suggested because inhibition or promotion of Ins(1,4,5)P(3) accumulation by neomycin or LiCl affects elicitor-induced production of beta-thujaplicin. Moreover, ruthenium red inhibits elicitor-induced accumulation of beta-thujaplicin while thapsigargin alone induces beta thujaplicin accumulation. These results suggest that Ca(2+) released from intracellular calcium stores may mediate elicitor-induced accumulation of beta thujaplicin via an Ins(1,4,5)P(3) signaling pathway, since it is widely accepted that Ins(1,4,5)P(3) can mobilize Ca(2+) from intracellular stores. This work demonstrates an elicitor-triggered Ins(1,4,5)P(3) turnover, defines its enzymatic basis and regulation, and suggests a role for Ins(1,4,5)P(3) in elicitor-induced phytoalexin accumulation via a Ca(2+) signaling pathway. PMID- 14747949 TI - Inhibition of a ubiquitously expressed pectin methyl esterase in Solanum tuberosum L. affects plant growth, leaf growth polarity, and ion partitioning. AB - Two pectin methyl esterases (PMEs; EC 3.1.1.11) from Solanum tuberosum were isolated and their expression characterised. One partial clone ( pest1) was expressed in leaves and fruit tissue, while pest2 was a functional full-length clone and was expressed ubiquitously, with a preference for aerial organs. Potato plants were transformed with a chimeric antisense construct that was designed to simultaneously inhibit pest1 and pest2 transcript accumulation; however, reduction of mRNA levels was confined to pest2. The decrease in pest2 transcript was accompanied by up to 50% inhibition of total PME activity, which was probably due to the reduction of only one PME isoform. PME inhibition affected plant development as reflected by smaller stem elongation rates of selected transformants when compared with control plants, leading to a reduction in height throughout the entire course of development. Expansion rates of young developing leaves were measured simultaneously by two displacement transducers in the direction of the leaf tip (proximal-distal axis) and in the perpendicular direction (medial-lateral axis). Significant differences in leaf growth patterns were detected between wild-type and transgenic plants. We suggest that these visual phenotypes could be correlated with modifications of ion accumulation and partitioning within the transgenic plants. The ion-binding capacities of cell walls from PME-inhibited plants were specifically modified as they preferentially bound more sodium, but less potassium and calcium. X-ray microanalysis also indicated an increase in the concentration of several ions within the leaf apoplast of transgenic plants. Moreover, quantification of the total content of major cations revealed differences specific for a given element between the leaves of PME-inhibited and wild-type plants. Reduced growth rates might also be due to effects of PME inhibition on pectin metabolism, predominantly illustrated by an accumulation of galacturonic acid over other cell wall components. PMID- 14747950 TI - Peripapillary atrophy in the unilateral exfoliation syndrome. AB - PURPOSE: To compare the areas of peripapillary atrophy between fellow eyes in patients with unilateral exfoliation syndrome. METHODS: Topographic measurements for peripapillary atrophy and the optic nerve head with confocal scanning laser ophthalmoscopy, using the Heidelberg Retina Tomograph were performed on 45 nonglaucomatous patients with unilateral exfoliation syndrome. The area of peripapillary atrophy was divided into an alpha and a beta zone. The areas of alpha and beta peripapillary atrophy and their angular extents around the disc were measured. The frequency distributions of the alpha and beta areas and their largest radial extents at different positions around the disc were calculated. RESULTS: An alpha area was detected in 84% of the exfoliative and 89% of the nonexfoliative fellow eyes, and a beta area in 31% and 42% of eyes, respectively. The exfoliative and the fellow nonexfoliative eyes did not differ in the sizes of the alpha (0.43+/-0.46 vs 0.33+/-0.18 mm(2), P=0.68) and beta (0.14+/-0.30 vs 0.17+/-0.34 mm(2), P=0.96) areas of peripapillary atrophy. The angular extents, the locations of the largest radial extent, and the frequency distributions of the alpha and beta peripapillary atrophy areas were similar in fellow eyes. CONCLUSION: Exfoliation syndrome itself is not a risk factor for peripapillary atrophy. PMID- 14747951 TI - Marked reduction and distinct patterns of eye blinking in patients with moderately dry eyes during video display terminal use. AB - PURPOSE: To analyze the spontaneous eye-blink rate (SEBR) and blinking pattern in patients with moderately dry eyes during video display terminal (VDT) use. METHODS: The SEBR of 30 patients with dry eye syndrome (median age 44.8 years, range 18 - 67 years) was measured using a computer-based video analysis system. Only complete eye blinks were recorded during 10 min of conversation, during initial VDT use and again after a preceding work period of 30 min. RESULTS: In comparison with SEBR during conversation (mean value 16.8 blinks/min, standard deviation 16.8 blinks/min) the SEBR significantly decreased during initial VDT use (6.6+/-4.8; P<0.001) and remained significantly decreased during re measurement after 30 min (5.9+/-4.6; P<0.001). SEBR was not significantly different between initial VDT use and re-measurement after 30 min ( P=0.65). The SEBR was characterized by marked interindividual variability during conversation and VDT use. No significant correlation was found between SEBR (either during conversation or during VDT use), and the quality (tear break-up time) or quantity (Schirmer I test, Jones test) of the tear film. Typical blinking patterns were found to be (1). a relatively time-independent, irregular pattern, (2). an initial phase of 2-4 min with shorter inter-eye-blink intervals, and (3). alternating periods of higher and lower inter-eye-blink intervals. CONCLUSIONS: Reduction of the SEBR during VDT use is primarily determined by marked visual attention, resulting in an exacerbation of dry eye symptoms in predisposed humans. High interindividual variability and distinct patterns of eye blinking may be key factors in further clarification of VDT-eye interactions and develop individually designed solutions to prevent desiccation during VDT use. PMID- 14747952 TI - Trans-Tenon's retrobulbar triamcinolone infusion for diffuse diabetic macular edema. PMID- 14747953 TI - Glomus tumor of the lateral aspect of the knee joint. AB - BACKGROUND: We report a rare case of glomus tumor that occurred in the lateral joint of the knee. A 54-year-old man was referred to us with a 3-year history of lateral pain in the left knee and the diagnosis of lateral meniscus injury. Physical examination revealed a small trigger point localized just on the lateral joint space. Magnetic resonance imaging (MRI) showed a very small round mass in the subcutaneous tissue at the level of the lateral joint space. TREATMENT: Excisional biopsy was performed, and the histology was characteristic of a glomus tumor. The patient was asymptomatic immediately after the operation. At the 3 year follow-up after the operation, no recurrence was seen. PMID- 14747954 TI - Sternal metastasis as initial presentation of a unknown rectal cancer. PMID- 14747955 TI - Supraumbilical pyloromyotomy: comparison between intracorporeal and extracorporeal approaches. AB - We evaluated intracorporeal pyloromyotomy (ICP) as an alternative to extracorporeal pyloromyotomy (ECP) in infants with hypertrophic pyloric stenosis. From July 1994 to June 2002, 75 patients underwent ICP, and 29 patients underwent ECP through supraumbilical incisions. The medical charts were reviewed retrospectively with regard to operating time, time to return to full feeding, complications, and outcome. The average operating time in the ICP group was significantly longer than in the ECP group (56.4+/-13.6 vs. 48.7+/-16.3 min; p=.004). However, the average time to return to full feeding in the ICP group was significantly shorter than in the ECP group (2.1+/-1.6 vs. 2.9+/-1.9 days; p=.002). Extension of the skin and/or fascial incision was necessary in two of 75 patients in the ICP group, compared with 11 of 29 patients in the ECP group ( p<.0001). There were four (5.3%) wound infections and one (1.3%) mucosal perforation in the ICP group, compared with two (6.9%) wound infections in the ECP group. ICP allows patients to return to full feeding earlier than ECP does, resulting in a shorter hospital stay. ICP through a supraumbilical incision is superior to the ECP technique. PMID- 14747956 TI - Intratumoral abscess: an unusual complication of ventriculoperitoneal shunt infection. AB - INTRODUCTION: Ventriculoperitoneal shunts were routinely used in the past in children with posterior fossa tumors and hydrocephalus. They can, however, cause a multitude of problems. CASE REPORT: This report highlights a previously unencountered phenomenon of a pyogenic abscess forming within a posterior fossa ependymoma as a result of shunt infection. The shunt was exteriorized and the child treated with antibiotics before surgery was done. Only a partial excision of the tumor was possible, as the inflammatory response caused by the abscess had obliterated tissue planes. PMID- 14747957 TI - Serological cloning of cancer/testis antigens expressed in prostate cancer using cDNA phage surface display. AB - Serological cloning of tumor-associated antigens (TAAs) using patient autoantibodies and tumor cDNA expression libraries (SEREX) has identified a wide array of tumor proteins eliciting B-cell responses in patients. However, alternative cloning strategies with the possibility of high throughput analysis of patient sera and tumor libraries may be of interest. We explored the pJuFo phage surface display system, allowing display of recombinant tumor proteins on the surface of M13 filamentous phage, for cloning of TAAs in prostate cancer (PC). Control experiments established that after a few rounds of selection on immobilized specific IgG, a high degree of enrichment of seroreactive clones was achieved. With an increasing number of selection rounds, a higher yield of positive clones was offset by an apparent loss of diversity in the repertoire of selected clones. Using autologous patient serum IgG in a combined biopanning and immunoscreening approach, we identified 13 different TAAs. Three of these (NY-ESO 1, Lage-1, and Xage-1) were known members of the cancer/testis family of TAAs, and one other protein had previously been isolated by SEREX in cancer types other than PC. Specific IgG responses against NY-ESO-1 were found in sera from 4/20 patients with hormone refractory PC, against Lage-1 in 3/20, and Xage-1 in 1/20. No reactivity against the remaining proteins was detected in other PC patients, and none of the TAAs reacted with serum from healthy subjects. The results demonstrate that phage surface display combined with postselection immunoscreening is suitable for cloning a diverse repertoire of TAAs from tumor tissue cDNA libraries. Furthermore, candidate TAAs for vaccine development of PC were identified. PMID- 14747958 TI - Iodine-131 treatment and chromosomal damage: in vivo dose-effect relationship. AB - Although it is well known that radiation induces chromosomal aberrations, there is a lack of information on the in vivo dose-effect relationship in patients receiving iodine-131 treatment, and the results of previous studies are controversial. In this study, the sister chromatid exchange (SCE) method was employed to investigate acute and late chromosomal damage (CD) in the peripheral lymphocytes of 15 patients who received various doses of (131)I (259-3,700 MBq), either for thyrotoxicosis (TTX) or for ablation treatment in differentiated thyroid cancer (DTC). The SCE frequencies in cultured peripheral lymphocytes were determined before treatment (to assess basal SCE frequencies), on the 3rd day (to assess acute SCE frequencies) and 6 months later (to assess late SCE frequencies). The basal, acute and late SCE frequencies (mean+/-SD) were 3.19+/ 0.93, 10.83+/-1.72 and 5.75+/-2.06, respectively, in the whole group, and these values differed significantly from each other ( P<0.001). In order to perform a quantitative evaluation of the present data and a comparative analysis with the results of previous studies reported in the literature, we defined acute and late effects using a "damage ratio" (DR) and a "recovery ratio" (RR), based on the basal, acute and late data for individual patients. No statistically significant difference was found in the DR between DTC and TTX patients (76.4%+/-11.5% vs 67.6%+/-9.0%), while the mean RR was higher in TTX patients than in the DTC group (75.2%+/-24.4% vs 36.8%+/-13.7%). The DR on the 3rd day was not related to the administered (131)I dose in the whole group, but a negative correlation was found between the (131)I dose and the RR at the 6th month (r=-0.60, P=0.04). The best fit for this relationship was obtained by a linear-quadratic model, as y=104.89x 28.4x(2)+38.1 ( R(2)=0.51, P=0.04). On the other hand, comparative analysis with the results of previous studies with comparable sampling times revealed that the best fit for the relationships between the administered dose of (131)I and DR and RR were obtained with a linear-quadratic model (Y=alpha D+beta D(2)) rather than a linear one. However, there was an interesting difference in comparison with in vitro studies, in that we found the coefficient beta to have a negative value, suggesting the disappearance of damaged lymphocytes from the peripheral circulation in a dose-dependent manner following (131)I treatment. Further studies are therefore needed to clarify the effect of the negative beta value on the biological dosimetry approach in continuous internal low LET radiation, as in the case of (131)I treatment. PMID- 14747960 TI - Highlights of the annual congress of the European Association of Nuclear Medicine, Amsterdam 2003. PMID- 14747959 TI - Risk of second primary cancer following differentiated thyroid cancer. AB - Concerns remain over the risk of cancer following differentiated thyroid carcinoma and its causes. Iodine-131 ((131)I) and external irradiation are known to have potential carcinogenic effects. Thyroid carcinoma is a polygenic disease which may be associated with other malignancies. We investigated the incidence of second cancer and its aetiology in a cohort of 875 patients (146 men, 729 women) with differentiated thyroid carcinoma originating from Basse-Normandie, France. Cancer incidence was compared with that of the general population of the Departement du Calvados matched for age, gender and period. The cumulative proportion of second cancer was estimated using the life-table method. Factors that correlated with the risk of second cancer were studied using the Cox model. After a median follow-up of 8 years, 58 second cancers had been observed. Compared with general population incidence rates, there was an overall increased risk of second cancer in women [standardised incidence ratio (SIR)=1.52; P<0.01], but not in men (SIR=1.27; P>0.20). Increased risk related to cancers of the genitourinary tract (SIR=3.31; P<0.001), and particularly to cancer of the kidney (SIR=7.02; P<0.01). Multivariate analysis showed that age above 40 years (P<0.01) and a history of previous primary cancer (P<0.001) correlated with risk. In contrast, neither cervical irradiation nor cumulative activity of (131)I was related to the risk. These data confirm that women with differentiated thyroid carcinoma are at risk of developing a second cancer of the genitourinary tract and kidney. Only age and medical history of primary cancer before thyroid carcinoma are risk factors for second cancer. Common environmental or genetic factors as well as long-term carcinogenic effects of primary cancer therapy should be considered. PMID- 14747961 TI - MR imaging in the evaluation of isolated limb perfusion: a prospective study of 18 cases. AB - PURPOSE: To prospectively evaluate the use of MRI with dynamic sequences during isolated limb perfusion (ILP) for soft tissue sarcomas, an aggressive local treatment using very high-dose chemotherapy and tumor necrosis factor aimed at avoiding limb amputation. DESIGN AND PATIENTS: Twenty-six patients were referred for ILP over one and a half years; eight were excluded as the lesions were either too proximal or suspicious inflammatory changes without tumor were found on the initial MRI, or the vascular status was poor. The indications for ILP were: vessel nerve involvement (13), multiple lesions (8), tumor size (4) or the presence of pulmonary metastases (2). MRI was performed 1 and 2 months after ILP, immediately prior to surgery and histological analysis. The MR examinations included T1-weighted SE and fast SE T2-weighted fat-saturated sequences, as well as dynamic sequences (T1-weighted SE repeated six times every 40 s), displaying the maximum intensity slope in each pixel. RESULTS: The tumor had disappeared in three patients. One patient still had histologically proven isolated widespread tumor cells without a mass. The tumor size had increased in two patients. In six patients, the size of the tumor had not changed but it had become completely necrotic, with a thin wall. In three patients, after an initially good result MRI demonstrated that the tumor wall had become thickened from 1 to 2 months after ILP. Dynamic MRI was mainly useful during the initial examination, demonstrating two patients with inflammatory changes without tumor. Three amputations and a second ILP were proposed based on poor results. Conservative limb-sparing surgery was successful in the other cases. CONCLUSION: MRI proved valuable in demonstrating the variable responses to ILP. PMID- 14747962 TI - Osseous Kaposi sarcoma in an HIV-positive patient. AB - A case of osseous Kaposi sarcoma in a 35-year-old man is described. The patient (HIV-positive for 8 years) suffered from cutaneous Kaposi sarcoma and presented with right-sided chest pain. He underwent a chest CT scan that revealed three osteolytic lesions involving rib and vertebra with large soft tissue masses, without cutaneous lesions at these sites. CT-guided core needle biopsy led to a histological diagnosis of Kaposi sarcoma. PMID- 14747963 TI - [Methodological perspectives for the assessment of adaptation in chronic diseases, exemplified by asthma bronchiale in childhood]. AB - BACKGROUND: Regarding the measurement of psychosocial adaptation due to chronic diseases in childhood and adolescence, there is a shift from a more reductionist and biomedical oriented disease-model towards a more integrated, biopsychosocial view of chronic diseases. The three paradigms in measuring psychosocial adaptation (psychopathology, coping, health related quality of life) will be discussed at the example of corresponding empirical studies in children with asthma bronchiale. The psychopathology-oriented research emphasizes the risk of the induced psychopathological comorbidity, whereas the more coping-oriented paradigm primarily includes the dynamic process of the perceived stress and the corresponding coping efforts due to a chronic disease. The third paradigm, the quality of life paradigm, sets its main focus on the subjective view of the chronically ill subject. CONCLUSIONS: In future studies, all three paradigms- each measuring different aspects of psychosocial adaptation--should be simultaneously included to come to a more complex view of adaptation in chronic diseases in general and asthma bronchiale in particular. PMID- 14747964 TI - Variables explaining the duration of hospitalization in children under two years of age admitted with acute airway infections: does respiratory syncytial virus have a direct impact? AB - BACKGROUND: Duration of hospitalization is influenced by many factors. It is an important parameter for quality of care. So far it is unknown, whether respiratory syncytial virus (RSV) etiology itself contributes to the time in hospital. METHOD: Children under 2 years of age admitted with a lower respiratory tract infection in 3 hospitals (1 tertiary and 2 secondary centers) in northern Germany were included in an unmatched, hospital-based case-control study. Cases were children tested positive for RSV by multiplex RT-PCR. One control group consisted of children tested negative for RSV in the multiplex-RT-PCR and a second control group consisted of patients in whom no PCR was done. Since only 4 to 5% in the latter group and thus 2% of the study population were misclassified, this group could be involved in the analysis. RESULTS: The median days of hospitalization was 7 days; 9 days in the RSV-positive group, 8 and 6 days in the RSV-negative and non-PCR group, respectively. The time in hospital was diminishing over the 4 year observation period. Duration of hospitalization was best predicted by--young age, presence of an underlying condition, disease entity -being pneumonia or bronchiolitis, prematurity, earlier epidemiologic year and intercostal retractions. Not predictive were: RSV-etiology, center, duration of illness, wheezing, C-reactive protein level and consolidation on the chest x-ray (all on admission). CONCLUSIONS: RSV-etiology is influencing the duration of hospitalization only indirectly via its predilection for the very young age and certain underlying conditions--not by RSV itself. In spite of considerable variation of patient populations between secondary and tertiary care facilities and locally different care practices, the treatment center pre se did not influence the duration of hospitalization significantly. Further rationalization of treatment is, however, possible. PMID- 14747965 TI - [Measures for the assessment of pain in neonates as well as a comparison between the Bernese Pain Scale for Neonates (BPSN) with the Premature Infant Pain Profile (PIPP)]. AB - BACKGROUND: Neonate's expression of pain lacks the ability to report pain. Several pain measures exist to assess acute pain in term and preterm neonates. The aim of the present study is to compare them with respect to their validity and reliability. METHOD: Review of the literature and a description of the measures most often cited. Additionally, the validity of the Bernese Pain Scale for Neonates (BPSN) was assessed in a department of neonatology of a university hospital. PATIENTS: Assessments of pain (n = 48) in term and preterm neonates with and without respiratory support. RESULTS: Existing pain measures are using behavioural indicators of pain (eg, facial expression, body posture, movements, and vigilance) as well as physiological indicators of pain (eg, changes in heart rate, respiratory rate, blood pressure, oxygen saturation). The used measures and their feasibility in everyday practice, the study population and the method of validation were presented. The BPSN differentiates pain from nonpain (F = 41.27, p < 0.0001) and the interrater- as well as the intrarater-reliability was high (r = 0.87 - 0.98 and r = 0.98 - 0.99, respectively). CONCLUSIONS: Assessment of acute pain in neonates should take into account the way of validation that has been performed especially with respect to the study population. The BSN is a pain measure with good validity and reliability for the assessment of pain in term and preterm neonates. PMID- 14747966 TI - [Linezolid in two premature babies with necrotizing enterocolitis and infection with vancomycin-resistant enterococcus]. AB - The use of the antibiotic Linezolid in two premature babies with necrotizing enterocolitis and infection with vancomycin-resistant enterococcus is described. Therapy with Linezolid in cause of necrotizing enterocolitis is an exception. Actually there are no proven recommendations for the use in this age group. Despite the successfully results more investigations for the use of linezolid in newborns and prematures are necessary. PMID- 14747967 TI - Failure to thrive in a 14-month-old boy with lymphopenia and eosinophilia. AB - We describe the case of a 14-month-old boy with delayed-onset SCID due to ADA deficiency which was masqueraded only by failure to thrive. Remarkably, the child had no serious infections and an adequate immune response. However, absolute lymphopenia, eosinophilia and absent thymus on chest x-ray were indicative for immunodeficiency. PMID- 14747968 TI - [Infant botulism and sudden infant death syndrome]. AB - Infant botulism represents a distinct entity of botulism. Ingestion of the ubiquitously present spores of Clostridium botulinum leads to germination of the organism and neurotoxin production in the infant intestine. Symptoms typically develop gradually in contrast to classical food botulism in which an acute onset of symptoms shortly after the ingestion of preformed toxin in a food is characteristic. Microbiologically, the diagnosis is established by identification of Clostridium botulinum organism and toxin in stool specimen. However, positive results in these tests provide only indirect evidence for the clinical relevance of the neurotoxin since asymptomatic carriers have been found. The toxin irreversibly blocks the release of acetylcholin from the motoric end plate which results in muscle weakness and paralysis. Depending on the amount of toxin produced, infant botulism exhibits a broad clinical spectrum ranging from oligosymptomatic forms to a fulminant course with acute respiratory failure within hours leading to sudden death. Unrecognized mild forms or beginning muscle weakness can be a co-factor for other risk factors of sudden infant death (SIDS). In studies analyzing infants who died from SIDS, botulism bacteria or toxin were found in up to 20 % of cases. Infant botulism therefore represents an important differential diagnosis of unexplained and inconclusive muscular hypotonia in the first year of life. PMID- 14747969 TI - [Sudden death of twins: botulism because of contamination by pap vegetables]. AB - Botulism is caused by the blockage of the neural transmission in the cholinergic synapses by botulinum neurotoxin (BoNT) which is produced by Clostridium botulinum or other Clostridia. The classic form of botulism occurs after the ingestion of food contaminated by BoNT. The course of the infection can be asymptomatic, mild with subtle paralysis ("failure to thrive") oder severe with generalized paralysis ("floppy infant"). Infected infants can also die sudden and unexpectedly. These deaths often are attributed to Sudden Infant Death Syndrome (SIDS), unless a thorough postmortem examination reveals Botulism. The rate of fatal Botulism falsely attributed to SIDS is not known, because it is difficult in most cases to show the causal relationship between contamination, disease and death. We report the sudden and unexpected simultaneous death of twins of 22 months which could be attributed to Botulism. Contamination of food, colonization of the gut by Clostridia and infection with specific pathomorphological changes could be proven. The initial suspicion of infanticide could be excluded. lt could be shown, that Botulism is a potential cause of simultaneous unexpected deaths in twins. PMID- 14747970 TI - [Doppler echocardiographic investigation of left ventricular diastolic function in preterm infants with and without a patent ductus arteriosus]. AB - We studied the influence of preload augmentation on Doppler-derived left ventricular diastolic function parameters in infants with a birth weight <1500 g. In 44 neonates with a closed duct and 29 neonates with an open ductus arteriosus a complete echocardiographic study including Doppler investigations of the mitral inflow signals and analysis of diastolic time intervals was performed. Neonates with an open duct had a lower median gestational age (27.1 vs. 29, P<0.01), a higher cardiac index (425 vs. 260 ml/min/kg, P<0.001), and a different M-mode left atrial to aortic diameter ratio (1.36 vs. 0.79, P<0.001), but birth weight, age at examination (6.6 vs. 7.9 days), and heart rate were similar compared to the neonates with a closed duct. Main differences in diastolic indexes existed in early and atrial filling integrals and peak velocities, early filling acceleration time, and isovolumic relaxation time. Most informative in neonates with an open ductus peak early filling velocity (41.2 vs. 30.4 cm, P<0.01) and peak atrial filling velocity (49.2 vs. 35.9, P<0.001) are higher, and isovolumic relaxation time is shorter (45 vs. 53 ms, P<0.001) than in closed duct peers. Isovolumic relaxation time is inversely correlated with the cardiac index (R = 0.78). These parameter changes can be interpreted as incipient left ventricular diastolic failure in extremely low birth weight infants in the presence of a preload challenge. The coincidence of higher peak velocities with a shortened isovolumic relaxation time is very likely a result of left atrial pressure elevation. Preload mismatch has to be considered in preterm infants with a persistent ductus arteriosus. The results of this study can be helpful to find the indication for PDA-closure and to adjust volume replacement therapy, and catecholamine medication to a level appropriate for the individual cardiac performance. PMID- 14747971 TI - Angiodysplasia as a cause of gastrointestinal bleeding in childhood. AB - Whereas in adults angiodysplasia is a frequent cause of gastrointestinal bleeding, in children this disorder is extremely rare. A 7 10/12 year old girl is presented suffering over 3-4 months from mild but recurrent rectal bleeding. Blood count and serum ferritin and transferrin levels were normal. The rectosigmoideoscopy revealed a rectal lesion, which was confirmed histologically as angiodysplasia. Pathological investigation of the biopsies included HE staining and immunohistological staining of endothelial cells with anti-CD34 and anti-von Willebrand factor. A follow-up period of three years revealed spontaneous regression of the angiodysplastic lesion at the rectosigmoideal localisation, which could be confirmed by endoscopy. CONCLUSION: The outcome of the few pediatric patients described in the literature was reviewed. Due to the lack of conclusive understanding of the nature of this extremely rare vascular disorder and the variable outcome described, a wait and see attitude should be assumed in cases of less clinical affection. PMID- 14747972 TI - Analysis of genetic relationships and antimicrobial susceptibility of Escherichia coli isolated from Clethrionomys glareolus. AB - Eleven strains of Escherichia coli were isolated from 54 bank voles living in the LomZa Landscape Park of the Narew River Valley, indicating that E. coli is not common in the alimentary tract of these mammals. On the basis of pulsed-field gel electrophoresis and computer-assisted analysis, the isolates were grouped into six genotypes at similarities of 39%. Chromosome length of E. coli under study differed by as much as 900 kb, ranging 2.7-3.6 Mb. All strains were susceptible to amikacin and ciprofloxacin, whereas, for tetracycline, streptomycin, ampicillin, and cefonicid, different results were noted. No differences were detected among the plasmid complements of eight strains (73%), for which plasmid profiles revealed the presence of two plasmidic bands. One, three and four plasmids were observed in a plasmid pattern of single isolates. The observation from the study indicated the high genetic polymorphism among the isolates recovered from the animals of one species living in the same environment. PMID- 14747973 TI - Role of some fermentation parameters on cyclosporin A production by a new isolate of Aspergillus terreus. AB - A local isolate of Aspergillus terreus was selected among different microorganisms as a new cyclosporin A (Cy A) producing culture. The formation of Cy A was investigated under different fermentation conditions (including selection of the cultivation medium, fermentation time course, inoculum nature, medium volume, agitation rate, pH value). Relatively high Cy A productivities were maintained when the fermentation process was carried out using a medium composed of (g/L): glucose, 50; bactopeptone, 10; KH(2)PO(4), 5; KCl, 2.5; pH 5.3, inoculated with 2% standard inoculum of 48 h age, shaken at 200 rpm for 10 days. PMID- 14747974 TI - Amino acids as main substrates for sulfate-reducing bacteria in surface sediment of a eutrophic bay. AB - The inner part of Tokyo Bay, Japan, is highly eutrophicated as shown by the frequent occurrence of red tide. The bottom water is anoxic during warm seasons especially at artificially dredged sites. In the sediment slurries prepared from surface sediment samples collected from the dredged sites, substrate addition stimulated the consumption of sulfate during anaerobic incubation. Of the substrates added, the seston composed mainly of diatom stimulated consumption more than lactate and acetate. Its effect was nearly equal to that of casamino acids. Casamino acids and some amino acids also accelerated the rate of sulfate reduction measured by the tracer method in sediment samples more than lactate or acetate. Anaerobic incubation of the sediment slurry amended with casamino acids showed that the consumption of amino acids was retarded by the addition of molybdate (final concentration; 20 mM). In the slurry amended with only molybdate, glutamate was accumulated distinctively and linearly with time. Its accumulation rate in molar base was comparable to the rate of sulfate reduction. These results suggested that amino acids were the main substrates for sulfate reducing bacteria (SRB) in the sediment. The MPN values of SRB in these sediment samples were often higher with the enumeration medium containing casamino acids instead of lactate. Furthermore, during a week incubation of sediment slurries amended with substrates, casamino acids and seston more greatly stimulated the growth of SRB enumerated by both media than lactate. PMID- 14747975 TI - Sporobolomyces magnisporus sp. nov., a new yeast species in the Erythrobasidium cluster isolated from plants in Taiwan. AB - Two strains of yeasts that contain Q-10 as the major ubiquinone, lack cellular xylose and produce large bilaterally symmetrical ballistoconidia were isolated from plants collected in a protected subtropical rain forest in Taiwan and were found to represent a new species. The taxonomic properties of this species coincide with the genus Sporobolomyces, so it is described as Sporobolomyces magnisporus sp. nov. In phylogenetic trees based on the nucleotide sequences of 18S ribosomal DNA and D1/D2 domain of 26S ribosomal DNA, this species is located in the Erythrobasidium cluster. PMID- 14747976 TI - Phylogenetic analyses of Lampropedia hyalina based on the 16S rRNA gene sequence. PMID- 14747977 TI - Cometabolic biodegradation of quinoline's derivatives by a quinoline-degrading bacteria: Comamonas sp. strain Q10. PMID- 14747978 TI - Banana puree fermentation by Lactobacillus acidophilus immobilized in Ca alginate. PMID- 14747979 TI - The UL24 consensus regions of bovine herpesvirus 2 isolate 554 and clinical isolates of herpes simplex viruses 1 and 2. PMID- 14747980 TI - [50 years of the Journal of psychosomatic medicine]. AB - This article is a review of the historical development of this journal from its establishment in 1954 to the present. The history of its founding will first be briefly outlined and illustrated with some facsimile prints from that time. The thematic focuses as they evolved over the last five decades will then be reviewed. In a further chapter individuals will be presented who have contributed significantly to the journal and made it what it is today. PMID- 14747981 TI - [Psychosomatic medicine as a scientific approach for the 21st century]. AB - OBJECTIVES: Subject of the article is the future of psychosomatic medicine as a science in the 21st century. METHODS: The state of the art is reviewed from the perspective of philosophy of science and sociology. The subjects of psychosomatic research are dealt with as well as theory and research methodology. RESULTS: Psychosomatic medicine will be influenced by a decrease in concrete interpersonal interaction and an increase in interaction directed by electronic media of communication. Holistic theories will be replaced by a variety of consistent and interdisciplinary informed middle range theories. And, last but not least, naive research concepts of the subjects of psychosomatic research will be supplemented by more complex concepts due to the multiplicity of qualitative and quantitative aspects of psychosomatic phenomena. DISCUSSION: The theoretical approach developed in this article will be illustrated by concrete examples from some research projects, in particular a project on the psychology of donors in living donor liver transplantation. PMID- 14747982 TI - [Results of psychodynamic inpatient psychotherapy in relation to diagnosis]. AB - Therapy outcome is analysed according to the main diagnoses in a 5 years' sample of psychodynamic inpatient psychotherapy (n=461) and controlled in a 6-9 months follow-up (n=312). Therapy effects, as measured in a pre-post comparison of the patients' self-rating, are generally good and especially noticeable for affective and anxiety disorders, whereas the effects for somatoform disorders are relatively low. The opposite tendencies are seen in follow-up, i.e. relapses in anxiety and affective disorders, and further improvement in somatoform disorders. A surprisingly positive outcome is shown for personality disorders and especially for severe personality disorders (e.g. Borderline). Regardless of the diagnoses, 80% of the patients show a high degree of satisfaction with their treatment and its outcome. PMID- 14747983 TI - [Impact of an additional behavioral anxiety intervention on therapy outcome in a psychodynamic inpatient setting]. AB - This naturalistic study aimed at evaluating the implementation of an additional symptom-oriented intervention program for patients with panic disorder and/or agoraphobia in a psychodynamic inpatient setting. Patients receiving this intervention showed a significantly better therapy outcome as measured by the symptom-specific scales of the SCL-90-R compared to anxiety patients who had not undergone this additional treatment. This specific improvement was stronger than the general improvement found in therapy outcome over time. Our findings show that the combination of different treatment strategies in one setting may prove effective. However, some problems are discussed which may emerge in clinical practice. PMID- 14747984 TI - [The relationship between OPD features of personality structure and symptom related and interpersonal outcome of inpatient psychotherapy]. AB - OBJECTIVE: To investigate the prognostic relevance of the axis "structure" of operationalized psychodynamic diagnostics (OPD) with respect to the symptom related and interpersonal outcome of inpatient psychotherapy by applying group comparisons as well as individual criteria. METHODS: The structural level of 102 psychiatric inpatients was related to the general and dissociative psychopathology (SCL-90; FDS-20) as well as interpersonal problems (IIP) at the beginning and at the end of inpatient treatment. RESULTS: There was a tendency toward an association between the structural level and psychopathological improvement. However, we did not find any clear relationship between structure and interpersonal difficulties either at the beginning or during the course of the treatment. The structural dimensions did not correlate with the reduction of psychopathology or interpersonal problems. From the patients perspective, however, overall personality structure, defence and object perception were important predictors for therapy success. CONCLUSIONS: In correspondence with other studies our findings suggest that consistent predictions on therapy outcome in regards to psychopathology and interpersonal problems are not possible on the basis of the axis "structure". The possible implications for a differential therapy indication are discussed. PMID- 14747985 TI - [How do patients experience their body after simultaneous pancreas-kidney transplantation? ]. AB - Simultaneous transplantation of pancreas and kidney (SPK) has become a standard procedure for type 1 diabetics with terminal kidney failure. Patients often experience SPK as a dramatic event which mobilizes a complexity of affects such as anxiety, hope, and gratitude. Patients often have fantasies regarding their bodies after transplantation which are linked to massive affects. METHODS: We investigated 8 non-selected patients in the first year after SPK by narrative interviews to study their experiences of transplantation. The interviews were audiotaped, transcribed and analysed according to Grounded Theory and ideal type analysis. RESULTS: Our patients had many different private theories and fantasies regarding transplantation. Patients' experiences were centred around themes of adoption of the transplanted organs and the protection of identity and self, and could be grouped into five ideal type sets. CONCLUSIONS: Bodily fantasies of our patients can be interpreted as specific adaptations to coping requirements after transplantation. PMID- 14747986 TI - Solution structure of the highly acidic protein HI1450 from Haemophilus influenzae, a putative double-stranded DNA mimic. AB - The solution structure of the acidic protein HI1450 from Haemophilus influenzae has been determined by NMR spectroscopy. HI1450 has homologues in ten other bacterial species including Escherichia coli, Vibrio cholerae, and Yersinia pestis but there are no functional assignments for any members of the family. Thirty-one of the amino acids in this 107-residue protein are aspartates or glutamates, contributing to an unusually low pI of 3.72. The secondary structure elements are arranged in an alpha-alpha-beta-beta-beta-beta order with the two alpha helices packed against the same side of an anti-parallel four-stranded beta meander. Two large loops, one between beta1 and beta2 and the other between beta2 and beta3 bend almost perpendicularly across the beta-strands in opposite directions on the non-helical side of the beta-sheet to form a conserved hydrophobic cavity. The HI1450 structure has some similarities to the structure of the double-stranded DNA (dsDNA) mimic uracil DNA glycosylase inhibitor (Ugi) including the distribution of surface charges and the position of the hydrophobic cavity. Based on these similarities, as well as having a comparable molecular surface to dsDNA, we propose that HI1450 may function as a dsDNA mimic in order to inhibit or regulate an as yet unidentified dsDNA binding protein. PMID- 14747987 TI - Myosin flexibility: structural domains and collective vibrations. AB - The movement of the myosin motor along an actin filament involves a directed conformational change within the cross-bridge formed between the protein and the filament. Despite the structural data that has been obtained on this system, little is known of the mechanics of this conformational change. We have used existing crystallographic structures of three conformations of the myosin head, containing the motor domain and the lever arm, for structural comparisons and mechanical studies with a coarse-grained elastic network model. The results enable us to define structurally conserved domains within the protein and to better understand myosin flexibility. Notably they point to the role of the light chains in rigidifying the lever arm and to changes in flexibility as a consequence of nucleotide binding. PMID- 14747988 TI - Structural principles of leucine-rich repeat (LRR) proteins. AB - LRR-containing proteins are present in over 2000 proteins from viruses to eukaryotes. Most LRRs are 20-30 amino acids long, and the repeat number ranges from 2 to 42. The known structures of 14 LRR proteins, each containing 4-17 repeats, have revealed that the LRR domains fold into a horseshoe (or arc) shape with a parallel beta-sheet on the concave face and with various secondary structures, including alpha-helix, 3(10)-helix, and pII helix on the convex face. We developed simple methods to charactere quantitatively the arc shape of LRR and then applied them to all known LRR proteins. A quantity of 2Rsin(phi/2), in which R and phi are the radii of the LRR arc and the rotation angle about the central axis per repeating unit, respectively, is highly conserved in all the LRR proteins regardless of a large variety of repeat number and the radius of the LRR arc. The radii of the LRR arc with beta-alpha structural units are smaller than those with beta-3(10) or beta-pII units. The concave face of the LRR beta-sheet forms a surface analogous to a part of a Mobius strip. PMID- 14747989 TI - Metal-Induced reversible structural interconversion of human mitochondrial NAD(P)+-dependent malic enzyme. AB - Human mitochondrial NAD(P)+-dependent malic enzyme was strongly inhibited by Lu3+. The X-ray crystal structures indicated a structural change between the metal-free and Lu3+-containing enzymes (Yang Z, Batra R, Floyd DL, Hung HC, Chang GG, Tong L. Biochem Biophys Res Commun 2000;274:440-444). We characterized the reversible slow-binding mechanism and the structural interconversion between Mn2+ and Lu3+-containing human mitochondrial malic enzymes. When Lu3+ was added, the activity of the human enzyme showed a downward curve over time, similar to that of the pigeon enzyme. The rate of the transformation (k(obs)) from the initial rate to the steady-state rate increased hyperbolically with the concentration of Lu3+, suggesting the involvement of an isomerization step. Lu3+ had a much higher affinity for the isomerized form (K*(i,Lu (app)) = 4.8 microM) than that of the native form (K(i,Lu (app)) = 148 microM). When an excess of Mn2+ was added to the Lu3+-inhibited enzyme, assays of the kinetic activity showed an upward trend, indicating reactivation. This result also indicated that the reactivation was a slow process. Fluorescence quenching experiments confirmed that the Lu3+-induced isomerization was completely reversible. The dynamic quenching constants for the metal-free, Mn2+-containing, and Lu3+-containing enzyme were 3.08, 3.07, and 3.8 M(-1), respectively. When the Lu3+-containing enzyme was treated with excess Mn2+, the dynamic quenching constant returned to the original value (3.09 M(-1)). These results indicated that binding of Mn2+ did not induce any conformational change in the enzyme. The open form transformed to the closed form only after substrate binding. Lu3+, on the other hand, transformed the open form into a catalytically inactive form. Excess Mn2+ could replace Lu3+ in the metal binding site and convert the inactive form back into the open form. This reversible process was slow in both directions because of the same but opposite structural change involved. PMID- 14747990 TI - Substrate binding to mononuclear metallo-beta-lactamase from Bacillus cereus. AB - Structure and dynamics of substrate binding (cefotaxime) to the catalytic pocket of the mononuclear zinc-beta-lactamase from Bacillus cereus are investigated by molecular dynamics simulations. The calculations, which are based on the hydrogen bond pattern recently proposed by Dal Peraro et al. (J Biol Inorg Chem 2002; 7:704-712), are carried out for both the free and the complexed enzyme. In the resting state, active site pattern and temperature B-factors are in agreement with crystallographic data. In the complexed form, cefotaxime is accommodated into a stable orientation in the catalytic pocket within the nanosecond timescale, interacting with the enzyme zinc-bound hydroxide and the surrounding loops. The beta-lactam ring remains stable and very close to the hydroxide nucleophile agent, giving a stable representation of the productive enzyme substrate complex. PMID- 14747991 TI - Three acidic residues are at the active site of a beta-propeller architecture in glycoside hydrolase families 32, 43, 62, and 68. AB - Multiple-sequence alignment of glycoside hydrolase (GH) families 32, 43, 62, and 68 revealed three conserved blocks, each containing an acidic residue at an equivalent position in all the enzymes. A detailed analysis of the site-directed mutations so far performed on invertases (GH32), arabinanases (GH43), and bacterial fructosyltransferases (GH68) indicated a direct implication of the conserved residues Asp/Glu (block I), Asp (block II), and Glu (block III) in substrate binding and hydrolysis. These residues are close in space in the 5 bladed beta-propeller fold determined for Cellvibrio japonicus alpha-L arabinanase Arb43A [Nurizzo et al., Nat Struct Biol 2002;9:665-668] and Bacillus subtilis endo-1,5-alpha-L-arabinanase. A sequence-structure compatibility search using 3D-PSSM, mGenTHREADER, INBGU, and SAM-T02 programs predicted indistinctly the 5-bladed beta-propeller fold of Arb43A and the 6-bladed beta-propeller fold of sialidase/neuraminidase (GH33, GH34, and GH83) as the most reliable topologies for GH families 32, 62, and 68. We conclude that the identified acidic residues are located at the active site of a beta-propeller architecture in GH32, GH43, GH62, and GH68, operating with a canonical reaction mechanism of either inversion (GH43 and likely GH62) or retention (GH32 and GH68) of the anomeric configuration. Also, we propose that the beta-propeller architecture accommodates distinct binding sites for the acceptor saccharide in glycosyl transfer reaction. PMID- 14747992 TI - Characterization of a thrombomodulin binding site on protein C and its comparison to an activated protein C binding site for factor Va. AB - Activation of the anticoagulant human plasma serine protease zymogen, protein C, by a complex of thrombin and the membrane protein, thrombomodulin, generates activated protein C, a physiologic anti-thrombotic, anti-inflammatory and anti apoptotic agent. Alanine-scanning site-directed mutagenesis of residues in five surface loops of an extensive basic surface on protein C was used to identify residues that play essential roles in its activation by the thrombin thrombomodulin complex. Twenty-three residues in the protein C protease domain were mutated to alanine, singly, in pairs or in triple mutation combinations, and mutants were characterized for their effectiveness as substrates of the thrombin thrombomodulin complex. Three protein C residues, K192, R229, and R230, in two loops, were identified that provided major contributions to interactions with thrombin-thrombomodulin, while six residues, S190, K191, K217, K218, W231, and R312, in four loops, appeared to provide minor contributions. These protein C residues delineated a positively charged area on the molecule's surface that largely overlapped the previously characterized factor Va binding site on activated protein C. Thus, the extensive basic surface of protein C and activated protein C provides distinctly different, though significantly overlapping, binding sites for recognition by thrombin-thrombomodulin and factor Va. PMID- 14747993 TI - Inhomogeneous stability of bacteriorhodopsin in purple membrane against photobleaching at high temperature. AB - Heterogeneity in the state of bacteriorhodopsin in purple membrane was studied through temperature jump experiments carried out in darkness and under illumination with visible light. The thermal denaturation, the irreversible component of spectral change at high temperature, had two decay components, suggesting that bacteriorhodopsin in purple membrane has heterogeneous stability. The temperature dependence of kinetic parameters under illumination revealed that the fast-decay component gradually increased at above 60 degrees C, indicating that the proportion of unstable bacteriorhodopsin increased. Significant change in the visible circular dichroism (CD) spectra was observed in darkness in the same temperature range as the increase of the fast-decay component under illumination. Denaturation experiments for C-terminal-cleaved bacteriorhodopsin showed that the C-terminal segment had some effect on the structural stability of bacteriorhodopsin under illumination. Dynamic and static models of the inhomogeneous stability of bacteriorhodopsin in purple membrane are discussed on the basis of the results of the denaturation kinetics and the visible CD spectra. PMID- 14747994 TI - Functional interaction of 13 yeast SCF complexes with a set of yeast E2 enzymes in vitro. AB - SCF complexes are multi-subunit ubiquitin ligases that, in concert with the E1 and E2 ubiquitination enzymes, catalyze the ubiquination of specific target proteins. Only three yeast SCFs have been reconstituted and characterized to date; each of these ubiquitinates its target protein with the E2 Cdc34. We have reconstituted and purified 1 known and 12 novel yeast SCF complexes, and explored the ability of these complexes to function with 5 different purified E2 enzymes; Ubc1, Cdc34, Ubc4, Ubc8 and Ubc11. We have found that the ubiquitination of Sic1 by the reconstituted SCF(Cdc4) complex was specifically catalyzed by two of the five E2 enzymes tested in vitro; Cdc34 and Ubc4. We also show that at least eight of the purified SCF complexes clearly ubiquitinated their F-box proteins in vitro, lending support for a regulatory mechanism in which F-box proteins catalyze their own destruction. The autoubiquitination of each F-box was in some cases catalyzed only by Cdc34, and in other cases preferentially catalyzed by Ubc4. Ubc4 thus interacts with multiple SCFs in vitro, and the interactions among SCF and E2 components of the ubiquitination machinery may allow further diversification of the roles of SCFs in vivo. PMID- 14747995 TI - Is there a weak H-bond --> LBHB transition on tetrahedral complex formation in serine proteases? AB - The transformation of a weak hydrogen bond in the free enzyme into a low-barrier hydrogen bond (LBHB) in the tetrahedral intermediate has been suggested as an important factor facilitating catalysis in serine proteases. In this work, we examine the structure of the H-bond in the Asp102-His57 diad of serine proteases in the free enzyme and in a covalent tetrahedral complex (TC) with a trifluoromethylketone inhibitor. We apply ab initio quantum mechanical calculations to models consisting of a large molecular fragment of the enzyme active site, and the combined effect of the rest of the protein body and the solvation by surrounding bulk water was simulated by a self-consistent reaction field method in our novel QM/SCRF(VS) approach. Potential profiles of adiabatic proton transfer in the Asp102-His57 diad in these model systems were calculated. We conclude that the hydrogen bond in both the free enzyme and in the enzyme inhibitor TC is a strong ionic asymmetric one-well hydrogen bond, in contrast to a previous suggestion that it is a weak H-bond in the former and a double-well LBHB in the latter. PMID- 14747996 TI - Aromatic side-chain interactions in proteins: near- and far-sequence Tyr-X pairs. AB - In the present study, an extensive analysis of the aromatic Tyr-X interactions is performed on a data set of 593 PDB structures, X being Phe, His, Tyr, and Trp. The nonredundant Tyr-X pairs (2645) were retained and separated by both the residue distance in the sequence and the secondary structures they bridge. Similar to the Phe-X and His-X pairs, the far-sequence Tyr-X pairs (X partner > five apart in the sequence: 74%) show comparable secondary structures and conformers for either type of X partner, in contrast with the near-sequence Tyr-X pairs (26%). As the Phe-X pairs, the near-sequence Tyr-X pairs stabilize secondary structures, mainly the alpha- helices (positions 1, 3, and 4) and the beta-strands (position 2). Like the Phe-X and His-X pairs, most far-sequence Tyr X pairs (34%) bridge beta-strands and only 11% bridge helices. As for the Phe-X and the His-X pairs, the X partners of the far-sequence Tyr-X pairs are frequently "above" the tyrosine ring with tilted and normal rings, whereas the X partner of the near-sequence Tyr-X pairs gradually moves from the "aside" to the "above" location, together with a progressive decrease of normal and increase of parallel rings, respectively. Unlike the His-X pairs, the interactions of the hetroatom in Tyr-X pairs are only favored with a sequence position +4 and over, owing to the spatial accessibility of the heteroatom. PMID- 14747997 TI - The number of protein folds and their distribution over families in nature. AB - Currently, of the 10(6) known protein sequences, only about 10(4) structures have been solved. Based on homologies and similarities, proteins are grouped into different families in which each has a structural prototype, namely, the fold, and some share the same folds. However, the total number of folds and families, and furthermore, the distribution of folds over families in nature, are still an enigma. Here, we report a study on the distribution of folds over families and the total number of folds in nature, using a maximum probability principle and the moment method of estimation. A quadratic relation between the numbers of families and folds is found for the number of families in an interval from 6000 to 30,000. For example, about 2700 folds for 23,100 families are obtained, among them about 33 superfolds, including more than 100 families each, and the largest superfold comprises about 800 families. Our results suggest that although the majority of folds have only a single family per fold, a considerably larger number of folds include many more families each than in the database, and the distribution of folds over families in nature differs markedly from the sampled distribution. The long tail of fold distribution is first estimated in this article. The results fit the data for different versions of the structural classification of proteins (SCOP) excellently, and the goodness-of-fit tests strongly support the results. In addition, the method of directly "enlarging" the sample to the population may be useful in inferring distributions of species in different fields. PMID- 14747998 TI - Comparison of backbone dynamics of monomeric and domain-swapped stefin A. AB - Three-dimensional domain swapping has been observed in increasing number of proteins and has been implicated in the initial stages of protein aggregation, including that of the cystatins. Stefin A folds as a monomer under native conditions, while under some denaturing conditions domain-swapped dimer is formed. We have determined the backbone dynamics of the monomeric and domain swapped dimeric forms of stefin A by (15)N relaxation using a model-free approach. The overall correlation times of the molecules were determined to be 4.6 +/- 0.1 ns and 9.2 +/- 0.2 ns for the monomer and the dimer, respectively. In the monomer, decreased order parameters indicate an increased mobility for the N terminal trunk, the first and the second binding loops. At the opposite side of the molecule, the loop connecting the alpha-helix with strand B, the beginning of strand B and the loop connecting strands C and D show increased localized mobility. In the domain-swapped dimer, a distinctive feature of the structure is the concatenation of strands B and C into a single long beta-strand. The newly formed linker region between strands B and C, which substitutes for the first binding loop in the monomer, has order parameters typical for the remainder of the beta-strands. Thus, the interaction between subunits that occurs on domain swapping has consequences for the dynamics of the protein at long-range from the site of conformational change, where an increased rigidity in the newly formed linker region is accompanied by an increased mobility of loops remote from that site. PMID- 14747999 TI - Comparison of the transition state ensembles for folding of Im7 and Im9 determined using all-atom molecular dynamics simulations with phi value restraints. AB - Delineation of the structural properties of transition states is key to deriving models for protein folding. Here we describe the structures of the transition states of the bacterial immunity proteins Im7 and Im9 obtained by all-atom molecular dynamics simulations with phi value restraints derived from protein engineering experiments. This pair of proteins is of special interest because, at pH 7 and 10 degrees C, Im7 folds via an intermediate while Im9 folds with a two state transition. The structures of the transition states for Im7 and Im9, together with their radii of gyration and distances from the native state, are similar. The typical distance between any two members of the transition state ensemble of both proteins is large, with that of Im9 nearly twice that of Im7. Thus, a broad range of structures make up the transition state ensembles of these proteins. The ensembles satisfy the set of rather low phi values and yet are consistent with high beta(T) values (> 0.85 for both proteins). For both Im7 and Im9 the inter-helical angles are highly variable in the transition state ensembles, although the native contacts between helices I and IV are well conserved. By measuring the distribution of the accessible surface area for each residue we show that the hydrophobic residues that are buried in the native state remain buried in the transition state, corresponding to a hydrophobic collapse to a relatively ordered globule. The data provide new insights into the structural properties of the transition states of these proteins at an atomic level of detail and show that molecular dynamics simulations with phi value restraints can significantly enhance the knowledge of the transition state ensembles (TSE) provided by the experimental phi values alone. PMID- 14748000 TI - A pharmacophore docking algorithm and its application to the cross-docking of 18 HIV-NNRTI's in their binding pockets. AB - The docking of small molecules into the binding site of a target protein is an important but difficult step in structure-based drug design. The performance of a docking algorithm is usually evaluated by re-docking ligands into their native binding sites. We have explored the cross-docking of 18 HIV-NNRTIs (non nucleoside inhibitors of HIV reverse transcriptase) of which the ligand-protein structure has been determined: each of the 18 ligands was docked into each of the 18 binding sites. The docking algorithms studied are an energy-based simulated annealing algorithm and a novel pharmacophore docking algorithm. It turns out that the energy-based docking of the ligands into non-native pockets is far less successful than the docking into their native pockets. The results can be improved by using explicit pharmacophore information, and by docking a ligand into a panel of protein structures and selecting the ligand-protein combination with the lowest interaction energy as the final result. PMID- 14748001 TI - Predicting peptide binding to MHC pockets via molecular modeling, implicit solvation, and global optimization. AB - Development of a computational prediction method based on molecular modeling, global optimization, and implicit solvation has produced accurate structure and relative binding affinity predictions for peptide amino acids binding to five pockets of the MHC molecule HLA-DRB1*0101. Because peptide binding to MHC molecules is essential to many immune responses, development of such a method for understanding and predicting the forces that drive binding is crucial for pharmaceutical design and disease treatment. Underlying the development of this prediction method are two hypotheses. The first is that pockets formed by the peptide binding groove of MHC molecules are independent, separating the prediction of peptide amino acids that bind within individual pockets from those that bind between pockets. The second hypothesis is that the native state of a system composed of an amino acid bound to a protein pocket corresponds to the system's lowest free energy. The prediction method developed from these hypotheses uses atomistic-level modeling, deterministic global optimization, and three methods of implicit solvation: solvent-accessible area, solvent-accessible volume, and Poisson-Boltzmann electrostatics. The method predicts relative binding affinities of peptide amino acids for pockets of HLA-DRB1*0101 by determining computationally an amino acid's global minimum energy conformation. Prediction results from the method are in agreement with X-ray crystallography data and experimental binding assays. PMID- 14748002 TI - Prediction of protein relative solvent accessibility with support vector machines and long-range interaction 3D local descriptor. AB - The prediction of protein relative solvent accessibility gives us helpful information for the prediction of tertiary structure of a protein. The SVMpsi method, which uses support vector machines (SVMs), and the position-specific scoring matrix (PSSM) generated from PSI-BLAST have been applied to achieve better prediction accuracy of the relative solvent accessibility. We have introduced a three-dimensional local descriptor that contains information about the expected remote contacts by both the long-range interaction matrix and neighbor sequences. Moreover, we applied feature weights to kernels in SVMs in order to consider the degree of significance that depends on the distance from the specific amino acid. Relative solvent accessibility based on a two state model, for 25%, 16%, 5%, and 0% accessibility are predicted at 78.7%, 80.7%, 82.4%, and 87.4% accuracy, respectively. Three-state prediction results provide a 64.5% accuracy with 9%; 36% threshold. The support vector machine approach has successfully been applied for solvent accessibility prediction by considering long-range interaction and handling unbalanced data. PMID- 14748003 TI - Analysis of the products of genes encompassed by the theoretically predicted pathogenicity islands of Mycobacterium tuberculosis and Mycobacterium bovis. AB - Sequencing of the genomes of Mycobacterium tuberculosis and Mycobacterium bovis provides a unique opportunity to study the biology of these pathogens on the genomic level. The computational detection of anomalous gene clusters such as those encompassed by pathogenicity islands allows for a narrowing of the study into well-defined groups of genes. Pathogenicity islands of M. tuberculosis (strains H37Rv and CDC1551) as well as M. bovis genomes comprise a group of genes encoding proteins that have been shown to be of immunological importance. The cross-genomic comparison (M. tuberculosis vs M. bovis) resulted in the elucidation of unique proteins in M. tuberculosis. These proteins may play a significant role in the host recognition process. PMID- 14748004 TI - From analysis of protein structural alignments toward a novel approach to align protein sequences. AB - Alignment of protein sequences is a key step in most computational methods for prediction of protein function and homology-based modeling of three-dimensional (3D)-structure. We investigated correspondence between "gold standard" alignments of 3D protein structures and the sequence alignments produced by the Smith Waterman algorithm, currently the most sensitive method for pair-wise alignment of sequences. The results of this analysis enabled development of a novel method to align a pair of protein sequences. The comparison of the Smith-Waterman and structure alignments focused on their inner structure and especially on the continuous ungapped alignment segments, "islands" between gaps. Approximately one third of the islands in the gold standard alignments have negative or low positive score, and their recognition is below the sensitivity limit of the Smith Waterman algorithm. From the alignment accuracy perspective, the time spent by the algorithm while working in these unalignable regions is unnecessary. We considered features of the standard similarity scoring function responsible for this phenomenon and suggested an alternative hierarchical algorithm, which explicitly addresses high scoring regions. This algorithm is considerably faster than the Smith-Waterman algorithm, whereas resulting alignments are in average of the same quality with respect to the gold standard. This finding shows that the decrease of alignment accuracy is not necessarily a price for the computational efficiency. PMID- 14748005 TI - Efficient methods for filtering and ranking fragments for the prediction of structurally variable regions in proteins. AB - The prediction of protein 3D structures close to insertions and deletions or, more generally, loop prediction, is still one of the major challenges in homology modeling projects. In this article, we developed ranking criteria and selection filters to improve knowledge-based loop predictions. These criteria were developed and optimized for a test data set containing 678 insertions and deletions. The examples are, in principle, predictable from the used loop database with an RMSD < 1 A and represent realistic modeling situations. Four noncorrelated criteria for the selection of fragments are evaluated. A fast prefilter compares the distance between the anchor groups in the template protein with the stems of the fragments. The RMSD of the anchor groups is used for fitting and ranking of the selected loop candidates. After fitting, repulsive close contacts of loop candidates with the template protein are used for filtering, and fragments with backbone torsion angles, which are unfavorable according to a knowledge-based potential, are eliminated. By the combined application of these filter criteria to the test set, it was possible to increase the percentage of predictions with a global RMSD < 1 A to over 50% among the first five ranks, with average global RMSD values for the first rank candidate that are between 1.3 and 2.2 A for different loop lengths. Compared to other examples described in the literature, our large numbers of test cases are not self-predictions, where loops are placed in a protein after a peptide loop has been cut out, but are attempts to predict structural changes that occur in evolution when a protein is affected by insertions and deletions. PMID- 14748006 TI - Splice variants: a homology modeling approach. AB - Splice variants play an important role within the cell in both increasing the proteome diversity and in cellular function. Splice variants are also associated with disease states and may play a role in their etiology. Information about splice variants has, until now, mostly been derived from the primary transcript or through cellular studies. In this study information from the transcript and other studies is combined with tertiary structure information derived from homology models. Through this method we have determined that it is possible to effectively model splice variants. Forty models of splice variants for fourteen proteins were produced. Analysis of the models shows that deletions produce superior model validation values. Additions to sequences where there is little homology become increasingly difficult to model with increasing sequence length. Many of the splicing events are associated with post-translational modification either in the N-terminal region by changing the signal peptide or by affecting the number or availability of glycosylation sites. Often the alternative exon combinations are associated with loss or gain of whole structural units, as opposed to just changing small loop regions. Losing part of the secondary structure may destabilize neighboring parts of the same secondary structure. Detailed analysis is given of four biomedically relevant proteins (Beta-site Amyloid Precursor Protein Cleaving enzyme (BACE), Interleukin-4, Frataxin and Hereditary hemochromatosis protein) and their associated splice variant models. The visualization of these possible structures provides new insights about their functionality and the possible etiology of associated diseases. PMID- 14748007 TI - C.C. Li (1912-2003): his science and his spirit. PMID- 14748008 TI - Efficient simulation of P values for linkage analysis. AB - In many genetic linkage analyses, the P value is obtained through simulation since the underlying distribution of the test statistic is complex and unknown. However, this can be very computationally intensive. A "bootstrap/replicate pool" approach has been suggested that generates P values more efficiently in terms of computation by resampling sums from a small set of simulated replicates for each pedigree. The replicate pool idea has been successfully applied, but, to our knowledge, has never been theoretically studied. An entirely different method for increasing the computational efficiency of P value simulation is Besag and Clifford's sequential sampling method. We propose an algorithm which combines Besag and Clifford's method with the replicate pool method to efficiently estimate P values for linkage studies. We derive variance expressions for the P value estimates from the replicate pool method and from our proposed hybrid method, and use these to show that the hybrid estimator has a substantial advantage over the other methods in most situations. PMID- 14748009 TI - Gamma regression improves Haseman-Elston and variance components linkage analysis for sib-pairs. AB - Existing standard methods of linkage analysis for quantitative phenotypes rest on the assumptions of either ordinary least squares (Haseman and Elston [1972] Behav. Genet. 2:3-19; Sham and Purcell [2001] Am. J. Hum. Genet. 68:1527-1532) or phenotypic normality (Almasy and Blangero [1998] Am. J. Hum. Genet. 68:1198-1199; Kruglyak and Lander [1995] Am. J. Hum. Genet. 57:439-454). The limitations of both these methods lie in the specification of the error distribution in the respective regression analyses. In ordinary least squares regression, the residual distribution is misspecified as being independent of the mean level. Using variance components and assuming phenotypic normality, the dependency on the mean level is correctly specified, but the remaining residual coefficient of variation is constrained a priori. Here it is shown that these limitations can be addressed (for a sample of unselected sib-pairs) using a generalized linear model based on the gamma distribution, which can be readily implemented in any standard statistical software package. The generalized linear model approach can emulate variance components when phenotypic multivariate normality is assumed (Almasy and Blangero [1998] Am. J. Hum Genet. 68: 1198-1211) and is therefore more powerful than ordinary least squares, but has the added advantage of being robust to deviations from multivariate normality and provides (often overlooked) model-fit diagnostics for linkage analysis. PMID- 14748010 TI - Conditional multipoint linkage analysis using affected sib pairs: an alternative approach. AB - Recently, Liang et al. ([2001b] Genet. Epidemiol. 21:105-122) proposed a conditional approach to assess linkage evidence on the target region by incorporating linkage information from an unlinked (reference) region using allele shared IBD (identity-by-decent) from affected sib pairs. This is carried out by conditioning on the IBD sharing value at the estimated trait locus of the reference region. Since markers considered are typically non-fully informative, the IBD sharing at each marker needs to be estimated (or imputed). In this report, we propose an alternative approach to deal with the IBD sharing in the reference region. This new approach makes full use of the observed data without having to categorize the imputed IBD sharing as needed in Liang et al. ([2001b] Genet. Epidemiol. 21:105-122). We compare these two approaches by simulating data from a variety of two-locus models including heterogeneity, additive and multiplicative with either fully informative markers or non-fully informative markers. The performance of both approaches is quite comparable showing consistent estimates of the trait locus and key genetic parameters. PMID- 14748011 TI - Power of direct vs. indirect haplotyping in association studies. AB - Haplotype analysis is essential to studies of the genetic factors underlying human disease, but requires a large sample size of phase-known data. Recently, directly haplotyping individuals was suggested as a means of maximizing the phase known data from a sample. Haplotyping, however, is much more labor-intensive than indirectly inferring haplotypes from genotypes (genotyping). This study uses simulations to compare the power of each methodology to detect associations between a haplotype and a trait or disease locus under conditions of varying linkage disequilibrium. The relative power of haplotyping over genotyping in association studies increases with decreasing sample size, decreasing linkage disequilibrium, increasing [corrected] numbers of marker loci, and decreasing numbers of different haplotypes. In addition, the frequency of the haplotype of interest and the magnitude of its association with the disease affect the power. From a cost-benefit standpoint, genotyping would be favored with large multiplicative risks (relative risk of haplotype >2.5). If case numbers are limiting rather than cost, haplotyping would maximize the information obtained. At small haplotype frequencies (e.g., <0.05), haplotyping is relatively more efficient, but there is little absolute power to detect associations under either methodology. Given the much larger laboratory resources required for direct haplotyping, genotyping would probably be favored under most conditions, but this must be balanced against the unit costs associated with recruitment and phenotyping. In the context of multipurpose, prospective cohort studies (e.g., the UK Biobank study), there may be a general value in establishing a series of directly haplotyped individuals to serve as controls for a number of alternative studies. PMID- 14748012 TI - A trisomic transmission disequilibrium test. AB - Certain congenital disorders that are rare in the general population are quite common in individuals with trisomic conditions. For example, complete atrioventricular septal defect occurs in about 20% of individuals with Down syndrome, an approximately 500-fold increase in risk as compared to individuals without Down syndrome. Genetic variation on the chromosome involved in the trisomy may affect susceptibility to these trisomy-specific disorders. That is, increased dosage of a variant may be directly involved in increasing the risk of a disorder, or it may be indirectly involved by causing up- or downregulation of other genes. As in standard disomic gene-mapping, one can search for genes using linkage or association methods. Within association methods, one can consider case control methods or family-based control methods such as the transmission disequilibrium test (TDT). Most gene-mapping methods need to be substantially redesigned for use with trisomic data. In this paper, we present a "trisomic TDT", a statistical method of testing for nonrandom transmission of alleles from parents to trisomic children. We demonstrate the method on a dataset of parent child trios in which the child has Down syndrome. PMID- 14748013 TI - What SNP genotyping errors are most costly for genetic association studies? AB - Which genotype misclassification errors are most costly, in terms of increased sample size necessary (SSN) to maintain constant asymptotic power and significance level, when performing case/control studies of genetic association? We answer this question for single-nucleotide polymorphisms (SNPs), using the 2x3 chi(2) test of independence. Our strategy is to expand the noncentrality parameter of the asymptotic distribution of the chi(2) test under a specified alternative hypothesis to approximate SSN, using a linear Taylor series in the error parameters. We consider two scenarios: the first assumes Hardy-Weinberg equilibrium (HWE) for the true genotypes in both cases and controls, and the second assumes HWE only in controls. The Taylor series approximation has a relative error of less than 1% when each error rate is less than 2%. The most costly error is recording the more common homozygote as the less common homozygote, with indefinitely increasing cost coefficient as minor SNP allele frequencies approach 0 in both scenarios. The cost of misclassifying the more common homozygote to the heterozygote also becomes indefinitely large as the minor SNP allele frequency goes to 0 under both scenarios. For the violation of HWE modeled here, the cost of misclassifying a heterozygote to the less common homozygote becomes large, although bounded. Therefore, the use of SNPs with a small minor allele frequency requires careful attention to the frequency of genotyping errors to ensure that power specifications are met. Furthermore, the design of automated genotyping should minimize those errors whose cost coefficients can become indefinitely large. PMID- 14748014 TI - Testing for association with a case-parents design in the presence of genotyping errors. AB - Genotyping errors can create a problem for the analysis of case-parents data because some families will exhibit genotypes that are inconsistent with Mendelian inheritance. The problem with correcting Mendelian inconsistent genotype errors by regenotyping or removing families in which they occur is that the remaining unidentified genotype errors can produce excess type I (false positive) error for some family-based tests for association. We address this problem by developing a likelihood ratio test (LRT) for association in a case-parents design that incorporates nuisance parameters for a general genotype error model. We extend the likelihood approach for a single SNP to include short haplotypes consisting of 2 or 3 SNPs. The extension to haplotypes is based on assumptions of random mating, multiplicative penetrances, and at most a single genotype error per family. For a single SNP, we found, using Monte Carlo simulation, that type I error rate can be controlled for a number of genotype error models at different error rates. Simulation results suggest the same is true for 2 and 3 SNPs. In all cases, power declined with increasing genotyping error rates. In the absence of genotyping errors, power was similar whether nuisance parameters for genotype error were included in the LRT or not. The LRT developed here does not require prior specification of a particular model for genotype errors and it can be readily computed using the EM algorithm. Consequently, this test may be generally useful as a test of association with case-parents data in which Mendelian inconsistent families are observed. PMID- 14748017 TI - Use of Kirschner wire in mandible reconstruction: a case report and review of the literature. AB - Reconstruction of massive defects in head the and neck usually requires different tissues such as skin, bone, and muscle in three-dimensional organization. Therefore, reconstruction of mandibular defects in particular include many difficulties for the head and neck, and for reconstructive surgeons. Various techniques and materials are available, and each has some advantages and disadvantages. But no materials or methods have all the reconstructive requirements. The surgeon's choice of techniques should be safe, simple, economic, and effective. We present a patient with a massive facial defect that was reconstructed successfully by using a 2-mm K wire with a free transverse rectus abdominis myocutaneous flap, and we also review the literature about the use of K wires in mandible fixation and reconstruction. We think that K wires should be kept in mind as a readily available, easily applicable, safe, inexpensive, and stable prosthetic material for mandible reconstruction, especially in cases with advanced disease. PMID- 14748015 TI - IL10 gene polymorphisms are associated with asthma phenotypes in children. AB - IL10 is an anti-inflammatory cytokine that has been found to have lower production in macrophages and mononuclear cells from asthmatics. Since reduced IL10 levels may influence the severity of asthma phenotypes, we examined IL10 single-nucleotide polymorphisms (SNPs) for association with asthma severity and allergy phenotypes as quantitative traits. Utilizing DNA samples from 518 Caucasian asthmatic children from the Childhood Asthma Management Program (CAMP) and their parents, we genotyped six IL10 SNPs: 3 in the promoter, 2 in introns, and one in the 3' UTR. Using family-based association tests, each SNP was tested for association with asthma and allergy phenotypes individually. Population-based association analysis was performed with each SNP locus, the promoter haplotypes and the 6-loci haplotypes. The 3' UTR SNP was significantly associated with FEV(1) as a percent of predicted (FEV(1)PP) (P=0.0002) in both the family and population analyses. The promoter haplotype GCC was positively associated with IgE levels and FEV(1)PP (P=0.007 and 0.012, respectively). The promoter haplotype ATA was negatively associated with lnPC(20) and FEV(1)PP (P=0.008 and 0.043, respectively). Polymorphisms in IL10 are associated with asthma phenotypes in this cohort. Further studies of variation in the IL10 gene may help elucidate the mechanism of asthma development in children. PMID- 14748018 TI - Antibiotics as an adjunct to surgical management of lower extremity ulcerations. PMID- 14748019 TI - Recent advances in the reconstruction of complex Achilles tendon defects. AB - Large, complex lower-extremity defects in the region of the Achilles tendon occur when tendon loss or disruption is complicated by damage to surrounding structures, including soft tissue, vessels, or bone. The surgical approach to these complex defects has evolved from simple amputation to the recognition that satisfactory reconstruction has three components: functional reconstruction of the tendon, importation of vascularized soft tissue, and skin coverage. Many techniques have been developed to address these difficult reconstructive goals, which often require multiple procedures or complicated single-stage operations. Microsurgical advances have begun to reduce the complexity of Achilles tendon region reconstruction, and excellent results can be obtained which restore function, form, and cosmesis with minimal morbidity. PMID- 14748020 TI - Aberrant position of the ulnar nerve within the carpal canal. AB - A case is presented where both the ulnar nerve and palmaris profundus tendon were found within the carpal tunnel during routine open decompression for carpal tunnel syndrome. PMID- 14748021 TI - Hyperbaric oxygen for the treatment of early-phase Dupuytren's contracture. AB - Dupuytren's disease (DD) is a proliferative disorder of autosomal-dominant inheritance, with variable penetrance of gene expression. The treatment of DD is challenging. Both operative and nonoperative approaches were reported for treatment of the disease, but no perfect approach has gained popularity as the best choice of treatment. Most of the emphasis has been placed on surgical techniques, but outcomes were reported to be dependent on some variables such as case selection, timing of surgery, and the surgeon's training and experience. In this paper, we report on a hyperbaric oxygen (HBO) treatment for early-phase DD. HBO treatment was applied to a female aged 23 years who had a mild form of DD. Physical findings and complaints before and after HBO treatment were compared. Total relief of symptoms as well as physical findings were obtained with HBO treatment. HBO for the treatment of DD is a novel concept. Having treated only one case is not enough to conclude that HBO is the only effective mode of treatment for DD. HBO should also be tried to treat early-phase or mild contractures of DD. Unfortunately, HBO has a disadvantage, i.e., cost. But HBO is not invasive, and because of the nature of HBO treatment, most of the complications seen after surgeries, e.g., wound-healing problems, damage to the digital nerves and vessels, buttonholing of the skin, and tendon sheath opening, are not seen. PMID- 14748022 TI - Long-term morphometric and immunohistochemical findings in human free microvascular muscle flaps. AB - Reinnervation, muscle regeneration, density of microvessels, and muscle-type specific atrophy were studied 3-4 years after surgery in surgically nonreinnervated free microvascular muscle flaps to 13 patients transplanted to the upper or lower extremities. Routine histology and immunohistochemistry for PGP 9.5 and S-100 (neuronal markers), Ki-67 (cell proliferation), myosin (muscle fiber types), and CD-31 (endothelium) were carried out, and results were analyzed morphometrically. Three to 4 years after surgery, severe atrophy of predominantly slow-type fibers was seen in 9 cases. In 4 cases, muscle-fiber diameter and fiber type distribution were close to normal. Long intraoperative muscle ischemia and postoperative immobilization were associated with poor muscle bulk in flaps. The density of microvessels in flaps did not differ from control muscles. PGP 9.5 and S-100 immunopositive nerve fibers were detected in 7 patients. Reinnervation was associated with good muscle bulk. In 4 patients, activation of satellite cells was evident. The results suggest that in some cases, spontaneous reinnervation may occur in free muscle flaps, and that several years after microvascular free flap transfer, the muscle still attempts to regenerate. PMID- 14748023 TI - Total thyroidectomy is improved by loupe magnification. AB - With this study, we verified if a microsurgical approach with magnification could improve the outcome of total thyroidectomy. Ninety-seven patients were consecutively randomized into group A (surgery with x 2.5 magnification and microsurgical instruments, n = 47) or group B (surgery with no magnification, n = 50). The mean operative time was 125 +/- 4.0 min in group A, and 150 +/- 4.0 min in group B (P = 0.00012). The recurrent laryngeal nerve was identified in all patients of group A, and in 96.8% of group B. The overall morbidity rate was 4.0% in group A and 25.5% in group B (P = 0.0038). This study indicates that a microsurgical approach with magnification is feasible, reduces surgical time, and improves the outcome in total thyroidectomy. PMID- 14748024 TI - Free anteromedial thigh flap: clinical application and review of literature. AB - The anteromedial thigh (AMT) flap is reviewed in terms of its vascular anatomy and previous clinical reports in the literature. Our own series of 5 patients treated with this flap for defects in the head and neck region and lower extremity is presented. Although several authors controversially discussed vasculature, we constantly found the pedicle as an emerging septocutaneous perforator at a point where the medial border of the rectus femoris muscle is crossed by the sartorius muscle. In all 5 patients, the AMT flap provided stable coverage with no flap loss. Based on our findings, we conclude that the anteromedial thigh flap offers all the advantages of fasciocutaneous flaps. Therefore, we recommend this flap as an alternative for defects requiring coverages of thin to moderate skin thickness. However, it should be remembered that variations in vascular anatomy are possible. PMID- 14748025 TI - Microsurgical penile replantation facilitated by postoperative HBO treatment. AB - Successful microsurgical replantation of a penis amputated at the level of the pubis is a rare occurrence worldwide. Moreover, the use of hyperbaric oxygen (HBO) for a postoperative replant Pseudomonas wound infection has not been reported. There is also disagreement regarding the importance of microsurgical repair of only the dorsal arteries or only the profundi arteries of the penis. A case is reported of penile replantation with a postoperative Pseudomonas wound infection treated with HBO to prevent potential replant loss, with a worldwide literature review. At 1-year postoperative follow-up, the patient has normal urinary flow and reports spontaneous erection, with the ability for intromission and a sensate glans. HBO facilitated the success of a penile replantation complicated by postoperative Pseudomonas wound infection. In addition, a literature review supports the microsurgical repair of at least a single isolated dorsal penile artery, but not a single or multiple profundi arteries. PMID- 14748026 TI - Emergency surgical repair of extensive soft-tissue defects in upper limb of a child, using flap graft. AB - Extensive soft-tissue defects in the upper limb of a child are rare, and emergency surgical repair presents a challenge to the clinician and requires high proficiency in operating skills. The use of common regular flaps fails to completely cover the trauma. We report on a case in which the skin defect, reaching 42 x 15 cm, was repaired by the use of a scapula latissimus dorsi musculo-cutaneous flap, and the functions of the affected limb were maintained to the utmost extent. Thus the patient has escaped amputation. PMID- 14748027 TI - Combined free-tissue transfer for primary reconstruction of radial part of the hand. AB - This paper presents 14 cases of primary reconstruction of radial-part hand defect by combined free-tissue transfer. The thumb was reconstructed by wraparound flap or second-toe transfer, while a lateral upper-arm flap or anterolateral thigh flap was transferred to cover the soft-tissue defect of the radial part of the hand. The arteries of the toe and flap were anastomosed and cascaded with the radial artery in an anatomical "snuffbox." Fourteen toes and 13 flaps survived in 14 cases. Only one flap was lost. Two-point discrimination of thumb tips was between 4-8 mm. The results suggest that combined free-tissue transfer appears to be a promising way of reconstructing a defect of the radial part of the hand. PMID- 14748028 TI - Compression neuropathy of common peroneal nerve caused by an extraneural ganglion: a report of two cases. AB - Peroneal nerve entrapment is most common in the popliteal fossa, but is rarely caused by a ganglion. Although ganglionic cysts are very common lesions, they seldom cause serious complications. Ganglionic cysts developing in the sheath of a peripheral nerve or joint capsule may cause compression neuropathy. We report on two cases of compression neuropathy of the common peroneal nerve caused by an extraneural ganglion and its evaluation with magnetic resonance imaging (MRI) and ultrasonography. The differential diagnosis should involve L5 root pathology, a posttraumatic intraneural hemorrhage, a nerve compression near the tendinous arch located at the fibular insertion of the peroneal longus muscle, and a nerve sheath tumor. The combination of MRI and ultrasonography is useful for the accurate diagnosis of this condition, and it should be treated by microsurgical exploration as soon as possible. PMID- 14748029 TI - Large recurrent plexiform neurofibroma of the foot and ankle. AB - Plexiform neurofibroma often causes significant pain, deformity, and functional problems in the affected part of the body. Surgical management of the condition can be challenging, and local recurrence is common because of inadequate resection of the tumor. One patient with a large recurrent plexiform neurofibroma of the foot and ankle was treated successfully with a complete resection of the tumor, followed by a free latissimus dorsi muscle transfer with a skin graft for soft-tissue reconstruction. The patient has regained good contour of the foot and ankle, and has been symptom-free with no evidence of local recurrence in the affected area for 18 months. The authors believe that thorough preoperative planning and state-of-the-art reconstructions, as described here, are the keys to their success. PMID- 14748030 TI - On sampling and sampling errors in histomorphometry of peripheral nerve fibers. AB - Histomorphometrical assessment of regenerated peripheral nerves is a very common goal of many studies in experimental microsurgery. In this paper, the main critical issues in nerve fiber sampling for quantitative morphological assessment are addressed. The equal opportunity rule, i.e., the basic paradigm of random sampling, is described, together with an explanation of how sampling errors, in the selection of histologic fields and of the nerve fibers inside them, can produce a bias in quantitative estimates. Finally, some practical suggestions on how to cope with the most common sampling errors are provided, in order to help researchers obtain reliable histomorphometrical data on peripheral nerve fibers. PMID- 14748031 TI - Arterialization of the portal stump by right renal artery after portocaval shunt in the rat. AB - Portosystemic shunts cause severe secondary effects, so that arterializations of the portal stump are planned to increase the blood supply to the liver. The aim of this study was to verify the technical feasibility of arterialization of the portal stump with the right renal artery in order to obtain a valid experimental model to study the pathophysiology of arterial revascularization of the liver. Twenty rats underwent end-to-side portocaval shunt + end-to-end anastomosis between the right renal artery and portal stump; another 20 rats were subjected to the same surgical procedure, but the portal stump underwent a reduction in size. In our study, both techniques gave good results, but the use of an operating microscope and good microsurgical training were essential in achieving a good patency rate (78.3%). PMID- 14748032 TI - Salutary effects of radiopaque contrast media on the survival of random-pattern skin flaps in the rat: an experimental study. AB - The radiopaque contrast medium diatrizoate, has a vasodilator effect so that it is used in sudden-deafness secondary ischemic injury. However, ischemic problems are encountered, especially when longer flaps are elevated. A longer flap also has ischemic and relatively ischemic tissue, and may obtain some benefit from contrast media. Forty male Sprague-Dawley rats, weighing about 350-400 g, were used, and randomly divided into four groups (n = 10 rats each group): group 1 was the control, group 2 the diatrizoate, group 3 the iopamidol, and group 4 the iothalamate group. A rectangular 3 x 10 cm caudally based dorsal skin flap was elevated, and sutured back to its original place. In the control group, no pharmacologic agent was administered. Sodium-meglumine-diatrizoate 10 mg/kg/day was administered parenterally in the first experimental group (diatrizoate group); iopamidol 10 mg/kg/day in the second experimental group (iopamidol group); and iothalamate sodium 10 mg/kg/day in the third experimental group (iothalamate group) for 7 postoperative days. On postoperative day 7, all flaps were photographed, and the area of flap survival was measured by using a polar planimeter. The results were statistically evaluated with the Kruskal-Wallis test and Mann-Whitney U-test (P = 0.05). The mean flap survival ranged from 79% in the iopamidol group to 83% in the diatrizoate group, and was significantly greater in all experimental groups (P < 0.05) compared to the control group (59%). There was no significant difference between experimental groups (P < 0.05). We believe that radiopaque contrast media have a beneficial effect in improving skin flap viability when distal flap necrosis is a potential complication of longer flaps. PMID- 14748033 TI - VEGF-promoted surgical angiogenesis in necrotic bone. AB - The ability of vascular endothelial growth factor (VEGF) to accelerate neoangiogenesis from implanted arterovenous (AV) bundles in necrotic bone was evaluated. A saphenous AV bundle was placed in a necrotic segment of rabbit ilium. In group II, VEGF (100 ng/h x 3 days) was administered by continuous infusion. Bone blood flow was measured with radioactive-labeled microspheres, and capillary density was determined by microangiography combined with Spalteholtz bone clearing at 1, 2, and 4 weeks. Neovascularization was observed along the implanted vascular bundle in both groups. One week after surgery, bone blood flow and vessel area were significantly higher in VEGF-treated animals (P < 0.05). No significant difference was observed at later times. Direct VEGF administration increased surgical angiogenesis and improved blood flow and neovascularization in necrotic bone 1 week after AV bundle implantation. Thereafter, a robust angiogenic response from the AV bundle was seen in both groups. PMID- 14748034 TI - Modelling clustered survival data from multicentre clinical trials. AB - In randomized clinical trials, subjects are recruited at multiple study centres. Factors that vary across centres may exert a powerful independent influence on study outcomes. A common problem is how to incorporate these centre effects into the analysis of censored time-to-event data. We survey various methods and find substantial advantages in the gamma frailty model. This approach compares favourably with competing methods and appears minimally affected by violation of the assumption of a gamma-distributed frailty. Recent computational advances make use of the gamma frailty model a practical and appealing tool for addressing centre effects in the analysis of multicentre trials. PMID- 14748035 TI - Bayesian methods for analysis of binary outcome data in cluster randomized trials on the absolute risk scale. AB - A Bayesian hierarchical modelling approach to the analysis of cluster randomized trials has advantages in terms of allowing for full parameter uncertainty, flexible modelling of covariates and variance structure, and use of prior information. Previously, such modelling of binary outcome data required use of a log-odds ratio scale for the treatment effect estimate and an approximation linking the intracluster correlation (ICC) to the between-cluster variance on a log-odds scale. In this paper we develop this method to allow estimation on the absolute risk scale, which facilitates clinical interpretation of both the treatment effect and the between-cluster variance. We describe a range of models and apply them to data from a trial of different interventions to promote secondary prevention of coronary heart disease in primary care. We demonstrate how these models can be used to incorporate prior data about typical ICCs, to derive a posterior distribution for the number needed to treat, and to consider both cluster and individual level covariates. Using these methods, we can benefit from the advantages of Bayesian modelling of binary outcome data at the same time as providing results on a clinically interpretable scale. PMID- 14748036 TI - Estimating linear regression models in the presence of a censored independent variable. AB - The current study examined the impact of a censored independent variable, after adjusting for a second independent variable, when estimating regression coefficients using "naive" ordinary least squares (OLS), "partial" OLS and full likelihood models. We used Monte Carlo simulations to determine the bias associated with all three regression methods. We demonstrated that substantial bias was introduced in the estimation of the regression coefficient associated with the variable subject to a ceiling effect when naive OLS regression was used. Furthermore, minor bias was transmitted to the estimation of the regression coefficient associated with the second independent variable. High correlation between the two independent variables improved estimation of the censored variable's coefficient at the expense of estimation of the other coefficient. The use of "partial" OLS and maximum-likelihood estimation were shown to result in, at most, negligible bias in estimation. Furthermore, we demonstrated that the full-likelihood method was robust under mis-specification of the joint distribution of the independent random variables. Lastly, we provided an empirical example using National Population Health Survey (NPHS) data to demonstrate the practical implications of our main findings and the simple methods available to circumvent the bias identified in the Monte Carlo simulations. Our results suggest that researchers need to be aware of the bias associated with the use of naive ordinary least-squares estimation when estimating regression models in which at least one independent variable is subject to a ceiling effect. PMID- 14748037 TI - Estimating life expectancy and related probabilities in screen-detected breast cancer patients with restricted follow-up information. AB - Issues such as life expectancy after diagnosis, the number of life years gained by early diagnosis through screening, the probability of dying from breast cancer or of dying from other causes during the lead time period or thereafter can be derived from information on complete survival after diagnosis. A method is presented to estimate complete survival and relevant outcomes after diagnosis of screen-detected cancer when the follow-up period is substantially shorter than the maximum follow-up possible. Survival after diagnosis until death from breast cancer is modelled as the sum of the lead time (LT) and the post-lead time (PLT), where both time periods follow exponential distributions and are assumed to be independent. The survival period after diagnosis until death from causes other than breast cancer (X) is assumed to follow a Gompertz distribution. The survival period after diagnosis until death from any cause (Z) is modelled as the minimum of LT+PLT and X. Maximum likelihood methods were then used to estimate all parameters of Z. This procedure for obtaining maximum likelihood estimates of Z does not need the cause of death (breast cancer or other), which is an advantage over most other methods. Especially in older patients, it may be difficult or even impossible to ascertain the true cause of death. The model was applied to data from the long-term breast cancer screening programme in Nijmegen, the Netherlands. Complete survival was estimated on the basis of survival data on 528 screen-detected breast cancer patients, diagnosed in 1975-1997 and with a mean follow-up of 8.9 years. Estimated life expectancy ranged between 22.3 and 9.0 years for patients diagnosed at the age of 50 and 79 years, respectively, that is, 6.1 and 0.7 life years gained by screening. Through early diagnosis and treatment, screen-detected patients diagnosed at the age of 50 years may have reduced their lifetime risk of dying from breast cancer from 79 per cent to 56 per cent; at the age of 79 the reduction of risk is reduced from 23 per cent to 13 per cent. PMID- 14748038 TI - Estimation of genetic and environmental factors for binary traits using family data. AB - While the family-based analysis of genetic and environmental contributions to continuous or Gaussian traits is now straightforward using the linear mixed models approach, the corresponding analysis of complex binary traits is still rather limited. In the latter we usually rely on twin studies or pairs of relatives, but these studies often have limited sample size or have difficulties in dealing with the dependence between the pairs. Direct analysis of extended family data can potentially overcome these limitations. In this paper, we will describe various genetic models that can be analysed using an extended family structure. We use the generalized linear mixed model to deal with the family structure and likelihood-based methodology for parameter inference. The method is completely general, accommodating arbitrary family structures and incomplete data. We illustrate the methodology in great detail using the Swedish birth registry data on pre-eclampsia, a hypertensive condition induced by pregnancy. The statistical challenges include the specification of sensible models that contain a relatively large number of variance components compared to standard mixed models. In our illustration the models will account for maternal or foetal genetic effects, environmental effects, or a combination of these and we show how these effects can be readily estimated using family data. PMID- 14748039 TI - Estimation of reporting delay and suicide incidence in Hong Kong. AB - This paper uses a semi-parametric method to examine the reporting delay distribution in suicides in Hong Kong reporting system. The data arise from a rightly truncated situation in which only suicide cases registered before a specific time are known to have occurred; otherwise they are not recorded in the known death files even if they have occurred. It is shown that the poisoning related suicide deaths have a longer reporting delay than other suicide methods. By modelling the reporting delay function, a Horvitz-Thompson-type estimator is suggested to adjust for reporting delay and to provide a more timely estimate of the suicide incidences for monitoring the suicide problem in Hong Kong. Based on these analyses, we recommended a suitable cut-off date to collect suicide cases occurring in the previous year and reported before this date in Hong Kong. PMID- 14748040 TI - Optimal designs for Michaelis-Menten kinetic studies. AB - Many reactions in enzymology are governed by the Michaelis-Menten equation. Characterising these reactions requires the estimation of the parameters K(M) and V(max) which determine the Michaelis-Menten equation and this is done by observing rates of reactions at a set of substrate concentrations. The choice of substrate concentrations is investigated by determining Bayesian D-optimal designs for a model in which residuals have a normal distribution with constant variance. Designs which focus on alternative quantities, such as K(M) or the ratio V(max)/K(M) are also considered. The effect on the optimal designs of alternative error distributions is also considered. PMID- 14748041 TI - A log-normal distribution model of the effect of bacteria and ear fenestration on hearing loss: a Bayesian approach. AB - Chronic ear infection is a potentially life-threatening illness that medical doctors typically treat with ear surgery. Despite the success of this treatment, complications can occur due to bacteria infection. Surgeons believe that this infection causes the patient to have clinically significant hearing damage. In order to understand such complications, surgeons must quantify the effect of bacteria, their toxins and ear surgery on hearing loss. To this end, the other two authors of this paper performed two experiments on guinea pigs to measure hearing thresholds following a bacterial infection and surgery of the inner ear. The response variable in these experiments is hearing thresholds measured in decibels (dB). The problem in analysing such experiments is that the hearing threshold observations often suffer from missing data and censoring mechanisms of various types. Additionally, the distribution of hearing thresholds has heavy tails and is peaked. In order to account for the above statistical issues, we present a Bayesian method with a location-shifted log-normal distribution. The method accounts for the uncertainty in the data collection mechanism and the parameters associated with a location-shifted log-normal distribution. We refer to one of the parameters as the "location-shift" parameter. The Bayesian approach provides a posterior distribution of the location-shift parameter that we compare with values estimated in previously published studies. The immediate goal of our proposed method was to quantify the effects of ear surgery and bacteria infection on hearing loss. Thus, we present the merits of the method in the form of a case study, and report posterior distributions of mean hearing loss, probability of clinically significant hearing loss and relative risk. The results show that surgeon 2, using the surgical procedure "oval window", poses a greater than 40 per cent chance of a 15dB hearing loss regardless of injection of bacteria or not. However, surgeon 1, using the surgical procedure "semicircular canal", does not pose a significantly greater than 40 per cent chance of a 15dB hearing loss unless there is a Pseudomonas aeruginosa-induced infection. PMID- 14748042 TI - Growth velocity assessment in paediatric AIDS: smoothing, penalized quantile regression and the definition of growth failure. AB - The analysis of growth records in paediatric anti-HIV clinical trials plays an important role in trial evaluation. Growth failure may be a manifestation of progressive disease or treatment toxicity, and is commonly specified as a major trial outcome event indicating poor treatment performance. Despite new therapeutic advances against HIV proliferation in infected patients, accurate monitoring and interpretation of somatic growth in paediatric AIDS remains clinically important, in light of uncertainties regarding relationship between viral load reductions and achievement of favourable somatic growth profiles. Our aim in this paper is to construct a criterion for growth failure that discriminates patients whose risk of death subsequent to growth failure is elevated to a clinically significant degree. To construct the criterion, individual growth curves and velocities are modelled using loess smoothing, penalized likelihood quantile regressions are fit to model age-specific growth velocity distributions for gender-stratified cohorts, and proportional hazards model selection is conducted to identify features of velocity series that are informative on the survival distribution. The resulting growth-failure criterion is expected to be useful for disease staging in resource-limited medical environments where T-cell counts and viral load measures are unavailable. PMID- 14748043 TI - Dr. T.K. Joshi and asbestos in India: a message from the Collegium Ramazzini. PMID- 14748044 TI - Quantitative exposure-response for silica dust and lung cancer in Vermont granite workers. AB - BACKGROUND: Excess lung cancer mortality among the exposed Vermont granite workers has been reported. These studies were based on job and tenure surrogates, with the potential for misclassification and inability to evaluate quantitative exposure-response. METHODS: Industrial hygiene data collected from 1924 to 1977 was analyzed in conjunction with mortality data to examine quantitative exposure response for silica, lung cancer, and other lung diseases. A person-years analysis was undertaken by cumulative exposure group, including lagged and unlagged tabulations. Poisson models were fitted to untransformed and log transformed exposure. RESULTS: The results indicated a clear relationship of lung cancer, tuberculosis, pneumoconiosis, non-malignant lung disease, and kidney cancer with cumulative exposure. An exposure to 0.05 mg/m(3) from age 20 to 64 was associated with a lifetime excess risk of lung cancer for white males of 27/1,000. CONCLUSIONS: The results of this study of workers exposed almost exclusively to silica and no other major occupational confounding exposures indicate a clear exposure-response for lung cancer. PMID- 14748045 TI - Work-related carpal tunnel syndrome (WR-CTS) in Massachusetts, 1992-1997: source of WR-CTS, outcomes, and employer intervention practices. AB - BACKGROUND: The Massachusetts Sentinel Event Notification System for Occupational Risks (MASS SENSOR) receives reports of work-related carpal tunnel syndrome (WR CTS) cases from (1) workers' compensation (WC) disability claims for 5 or more lost work days; and (2) physician reports (PR). METHODS: From 1992 through 1997, 1,330 WC cases and 571 PR cases completed follow-back surveys to provide information on industry, occupation, attributed source of WR-CTS, outcomes, and employer intervention practices. RESULTS: Sixty-four percent of the respondents had bilateral CTS and 61% had surgery, both of which were proportionally more frequent among WC cases. Office and business machinery was the leading source of WR-CTS (42% of classifiable sources) in every economic sector except construction, followed by hand tools (20%). Managers and professional specialty workers were the most likely to report employers' interventions and were up to four times more likely to report equipment or work environment changes than higher risk groups. CONCLUSIONS: State-based surveillance data on the source of WR-CTS provided valuable information on how and where to implement interventions. New occurrences of WR-CTS are likely, especially in the highest risk industries where very few cases reported primary prevention measures (e.g., changes to equipment or work environment) implemented by their employers. PMID- 14748046 TI - Plutonium-related work and cause-specific mortality at the United States Department of Energy Hanford Site. AB - BACKGROUND: Health effects of working with plutonium remain unclear. Plutonium workers at the United States Department of Energy (US-DOE) Hanford Site in Washington State, USA were evaluated for increased risks of cancer and non-cancer mortality. METHODS: Periods of employment in jobs with routine or non-routine potential for plutonium exposure were identified for 26,389 workers hired between 1944 and 1978. Life table regression was used to examine associations of length of employment in plutonium jobs with confirmed plutonium deposition and with cause specific mortality through 1994. RESULTS: Incidence of confirmed internal plutonium deposition in all plutonium workers was 15.4 times greater than in other Hanford jobs. Plutonium workers had low death rates compared to other workers, particularly for cancer causes. Mortality for several causes was positively associated with length of employment in routine plutonium jobs, especially for employment at older ages. At ages 50 and above, death rates for non-external causes of death, all cancers, cancers of tissues where plutonium deposits, and lung cancer, increased 2.0 +/- 1.1%, 2.6 +/- 2.0%, 4.9 +/- 3.3%, and 7.1 +/- 3.4% (+/-SE) per year of employment in routine plutonium jobs, respectively. CONCLUSIONS: Workers employed in jobs with routine potential for plutonium exposure have low mortality rates compared to other Hanford workers even with adjustment for demographic, socioeconomic, and employment factors. This may be due, in part, to medical screening. Associations between duration of employment in jobs with routine potential for plutonium exposure and mortality may indicate occupational exposure effects. PMID- 14748047 TI - Mortality patterns among workers exposed to styrene in the reinforced plastic boatbuilding industry: an update. AB - BACKGROUND: Mortality was updated through 1998 for 5,204 workers exposed to styrene between 1959 and 1978 at two reinforced plastic boatbuilding plants. The a priori hypothesis: leukemia and lymphoma excesses would be found. METHODS: Standardized mortality ratios (SMR) and 95% confidence intervals (CI) used Washington State and U.S. rates. RESULTS: Overall, 860 deaths occurred (SMR 1.09, CI 1.02-1.17), with excess mortality for esophageal cancer (n = 12, SMR 2.30, CI 1.19-4.02), prostate cancer (n = 24, SMR 1.71, CI 1.09-2.54), and accidents (n = 99, SMR 1.26, CI 1.02-1.53). Among 2,062 highly exposed workers, urinary tract cancer (n = 6, SMR 3.44, CI 1.26-7.50) and respiratory disease (n = 12, SMR 2.54, CI 1.31-4.44) rates were elevated. Urinary tract cancer SMR increased with duration of employment. CONCLUSIONS: We found no excess leukemia or lymphoma mortality. Unanticipated excess urinary tract cancer and respiratory disease mortality, possibly associated with styrene exposure, are difficult to interpret and could be chance findings. PMID- 14748048 TI - Fatal and non-fatal machine-related injuries suffered by children in Alberta, Canada, 1990-1997. AB - BACKGROUND: Children raised on farms are exposed to many hazardous types of machinery. The objective of this study was to describe the magnitude of non-fatal and fatal farm machinery injuries in Alberta children and adolescents (0-17 years) for the years 1990-1997. To date, there have been no published studies of pediatric farm injuries in Western Canada. METHODS: Data were collected through the Canadian Agricultural Injury Surveillance Program (CAISP). Death certificates and hospital charts were audited to provide enhanced information about the circumstances of injuries related to farm machinery for farm persons aged 17 years and younger. RESULTS: A total of 302 farm machinery injuries were recorded for the years 1990-1997. Of these, 14 resulted in death. All-terrain vehicles (ATVs) were the most common cause of injury (n = 76), followed by tractors (n = 72), and power take-offs (n = 15). The predominant injury mechanism was entanglement (n = 69), followed by falls from machines (n = 57), and being pinned/struck by a machine (n = 49). The median length of hospital stay for injuries was 2.0 days. Males (median = 2.0 days) had significantly longer hospital stays than females (median = 1.0 days). There were significantly more injuries reported during the summer and autumn than during the winter and spring. Those injured in the autumn were significantly older (median = 13.0 years) than children injured in the spring (median = 9.0 years). Injury rates dropped significantly during the study period from 119.9/100,000 per year in 1990 to 50.7/100,000 in 1997. CONCLUSIONS: While injury rates have dropped, the number of injuries occurring to children on Alberta farms is of concern. The large number of ATV related injuries suggests that preventative strategies need to be focused in this area. Am. J. Ind. Med. 45:177-185, 2004. PMID- 14748049 TI - Concentration of hydroxyproline in blood: a biological marker in occupational exposure to asbestos and its relationship with Pi*Z and Pi*S polymorphism in the alpha-1 antitrypsin gene. AB - BACKGROUND: Hydroxyproline (OHP) is one of the most abundant amino acids in collagen and, in general, it provides a good measure of overall collagen catabolism. METHODS: Asbestos workers suffering from asbestosis (cases n = 85); asbestos exposed workers without asbestosis (exposed controls, EC, n = 86), and non-exposed population (non-exposed controls, NEC, n = 122) were studied. The concentration of free OHP in whole blood was measured following the Pico-Tag procedure. RESULTS: Concentration of OHP in blood was significantly different in the three groups studied (P < 0.001), being higher in cases (19.8 +/- 14.7 micromol/L) than in EC (16 +/- 12.4) and NEC (13.5 +/- 6.7). When all individuals were grouped and stratified by the Pi*S and Pi*Z polymorphisms in the alpha-1 antitrypsin gene, the highest OHP levels were detected in the Pi*S homozygotes, one of the asbestosis-at risk-genotypes (Pi*S homozygotes, x = 24.5 +/- 11.7; Pi*S heterozygotes, x = 16.6 +/- 10.0; wild type, wt, x = 15.9 +/- 11.8). CONCLUSIONS: Blood OHP concentration could be used for monitoring human exposure to asbestos, either as a marker for occupational monitoring or as an additional clinical parameter in diagnostic exploration of asbestosis. PMID- 14748050 TI - High prevalence of silicosis among stone carvers in Brazil. AB - BACKGROUND: In the city of Petropolis, Brazil, artisans carve souvenirs from a variety of silica-containing minerals. The finding of pulmonary massive fibrosis in one of the workers motivated an investigation of the prevalence of silicosis in this group. METHODS: Between January 2000 and June 2002, a cross-sectional study was performed. We obtained clinical and occupational histories, spirometry, lung volumes, and carbon monoxide diffusion capacity measurements. Chest radiographs and high-resolution computed tomographies (HRCT) were evaluated. Personal air samples were analyzed. RESULTS: Forty-two stone carvers were examined. The prevalence of silicosis was 53.7%. HRCT better characterized silicotic lesions compared to chest radiographs. Early coalescence of small opacities was associated with lung function impairment. The concentration of dust exceeded permissible limits in 91% of the workplaces. CONCLUSIONS: Exposure to high levels of silica dust was associated with an increased prevalence of silicosis among stone carvers. Am. J. Ind. Med. 45:194-201, 2004. PMID- 14748051 TI - Osteoarthritis of the hip joint and farm work. AB - BACKGROUND: Different types of farming and farm work were studied in relation to the occurrence of osteoarthritis of the hip joint in a cohort of farmers. METHODS: A group of 427 farmers with hip joint osteoarthritis were identified together with a matched control group of farmers without hip or knee problems. All were invited to an interview and physical examination. After a dropout of 10%, and removing those who had had predisposing diseases or trauma, the observations for 369 pairs were available for analyses. RESULTS: Farmers with larger dairy and swine confinement operations (sows) had an increased risk of acquiring osteoarthritis of the hip. In a logistic regression multivariate model, those who milked more than 40 cows daily had an increase in risk (OR = 4.5, 95% CI 1.9-11.0) in relation to those who did not work in dairy production. Those who had worked more than 5 hr daily in animal barns over a long period of time also had an increase in risk (OR = 13.3, 95% CI 1.2-145.0) in relation to those who did not work with animals. In another model, those who had large farm areas (>100 ha) had a significantly decreased risk (OR = 0.14, 95% CI 0.05-0.43) in relation to those who had smaller farm areas. CONCLUSIONS: Farmers operating farms with large plant production area and few animal contacts had a significantly lower risk of incurring osteoarthritis of the hip than did farmers in general. The presence of animal production showed a significant positive relationship to the risk of developing hip joint osteoarthritis. Am. J. Ind. Med. 45:202-209, 2004. PMID- 14748052 TI - Effectiveness of the HomeSafe Pilot Program in reducing injury rates among residential construction workers, 1994-1998. AB - BACKGROUND: The construction industry typically has one of the highest fatal and non-fatal injury rates compared with other industries. Residential construction workers are at particular risk of injury (work is in remote sites with small crews, there are often many subcontractors, and they have limited access to safety programs). Difficulty accessing information specific to this group has made research more challenging, therefore, there are few studies. This study evaluated the effectiveness of the HomeSafe Pilot Program, a safety education and training program designed to reduce injuries among residential construction workers. METHODS: Researchers evaluated whether overall and severe injury incidence rates declined during the intervention period. Data were analyzed using incidence rates and Poisson regression to control for the effect of antecedent secular trend. RESULTS: Injury incidence rates declined significantly following HomeSafe; however, this effect was not statistically significant once temporal variation was controlled. CONCLUSIONS: The decline in injury rates following HomeSafe cannot be attributed solely to HomeSafe, however, programmatic and methodologic limitations contributed to the inconclusive results. Further research into the hazards faced by residential construction workers is needed. Am. J. Ind. Med. 45:210-217, 2004. PMID- 14748053 TI - Review of the Federal Child Labor Regulations: updating hazardous and prohibited occupations. AB - BACKGROUND: Child labor regulations are intended to protect our young and most vulnerable workers. While regulations are not the only strategy for providing a safe and healthy work environment for young workers, they are the first step in setting the community standard to protect them. The world of work has changed since the first child labor laws were issued in 1939. The U.S. Department of Labor (DOL) enlisted assistance from the National Institute for Occupational Safety and Health (NIOSH) to evaluate the current list of hazardous and prohibited occupations and to make recommendations for changes to the regulations. METHODS: A group of national experts reviewed the recommendations made by NIOSH. The consensus review process identified areas of agreement, disagreement, and priorities for the DOL's Wage and Hour Division. Gaps in the hazardous orders not addressed in the recommendations were also identified. RESULTS: This commentary describes the background of the Hazardous Orders under the Fair Labor Standards Act, the evaluation by NIOSH, the process used to review it, and recommendations for further action. CONCLUSIONS: The NIOSH report is a valuable resource for anyone working to prevent occupational injuries-among adult and young workers alike. It is crucial that the DOL use the guidance provided by NIOSH and the Young Workers Safety and Health Network to improve the protections for young workers. PMID- 14748054 TI - Leukemia risk and relevant benzene exposure period-Re: follow-up time on risk estimates, Am J Ind Med 42:481-489, 2002. PMID- 14748056 TI - Effects of potent antiretroviral therapy on the immune activation marker soluble CD27 in patients infected with HIV-1 subtypes A-D. AB - HIV-1 genetic subtypes might have a different impact on disease progression and response to antiretroviral therapy (ART). Few data are available on the immune activation profile in patients with different HIV-1 subtypes. We have tested by ELISA the plasma levels of an immune activation marker, soluble CD27 (sCD27), in a cohort of 64 patients infected with HIV-1 subtypes A-D, at baseline and after 1 year of virologically successful ART. Plasma sCD27 was significantly higher in the whole HIV-1-infected population as compared to healthy subjects [522 U/ml (188-1,307) vs. 285 U/ml (174-397), P < 0.001]. Among the four different HIV-1 subtypes, patients with subtype C virus had significantly higher plasma sCD27 [684 U/ml, (188-1228)] as compared to patients with subtype A [428 U/ml (247 1307), P < 0.05] and B [454 (211-925), P < 0.05]. After 1 year of ART, plasma sCD27 significantly decreased in all groups but patients with subtype C viruses had the largest reduction of sCD27 from baseline. The data indicate that a similar immune activation profile is present in patients infected with HIV-1 subtypes A, B, and D and that in presence of successful ART these subtypes respond similarly in terms of immune activation. Intriguingly, subtype C infection seems to be associated with higher levels of plasma sCD27, suggesting that HIV-1 genetic subtype C may have a different impact on disease outcome and response to therapy. PMID- 14748057 TI - Different resistance mutations can be detected simultaneously in the blood and the lung of HIV-1 infected individuals on antiretroviral therapy. AB - In this retrospective study, matched peripheral blood and lung samples from patients on antiretroviral therapy were studied in order to investigate whether differences in mutations associated with resistance to nucleoside analogues could be detected between the lung and blood. Discordant mutation patterns in the reverse transcriptase (RT) between plasma and cell free bronchoalveolar lavage fluid (BAL-fluid) HIV-1 genomic RNA was observed in five out of seven patients on nucleoside reverse transcriptase inhibitor (NRTI) monotherapy and six out of seven on combination therapy. In the cellular compartments, DNA recovered from peripheral blood mononuclear cells (PBMCs) and cells from BAL-cells discordant HIV-1 resistance genotypes were detected in 15 out of 44 matched samples. Differences in resistant genotypes between PBMCs and BAL-cells were most pronounced in patients receiving combination antiretroviral therapy. The pattern and number of mutations in RT associated with resistance differed in the BAL cells compared to PBMCs in four out of 12 subjects not receiving antiretroviral therapy at the time of bronchoscopy, three from 14 patients on NRTI monotherapy, five out of nine on dual combination therapy and three out of nine on HAART. The differences in the detection of resistance mutations between blood and the lung suggest that the lung is a site of replication for HIV-1. PMID- 14748058 TI - HIV epidemic in Central African Republic: high prevalence rates in both rural and urban areas. AB - A sentinel serosurveillance study was conducted in Central African Republic to estimate the prevalence of HIV seropositivity in the general adult population in each province so that the public health authorities can target HIV prevention programmes to the priority areas. Blood samples were collected from women attending 48 antenatal clinics in urban and rural areas of the Central African Republic. These samples were tested for HIV antibodies in an anonymous and unlinked manner using strategy II recommended by WHO. The data were extrapolated to all women of reproductive age in Central African Republic by use of a parity based adjustment involving the application of correction factors to the observed prevalence rates. A total of 9,305 pregnant women were recruited from November 2001 to October 2002. HIV seroprevalence was high in all age groups (12% in the less than 20 year age group to 17% in the 25-29 year age group). The median prevalence of HIV in antenatal clinics was similar for rural areas, for Bangui and for other urban areas (16.5, 15.0, and 12.5% respectively). Adjustment for parity and fertility pattern increased the prevalence of HIV in all antenatal clinics except in Bangui. This first national study of HIV prevalence in Central African Republic revealed that the HIV epidemic is continuing to spread in both urban and rural areas. Thus, efforts to reduce transmission should be made in every part of the country. PMID- 14748059 TI - Hepatitis B virus genotypes and spontaneous hepatitis B e antigen seroconversion in Taiwanese hepatitis B carriers. AB - Hepatitis B virus (HBV) is classified into eight genotypes (A-H), and genotype C is associated with more aggressive liver disease compared to genotype B. However, the mechanisms responsible for the clinical differences remain unclear. To test whether genotype C patients had with lower rates of spontaneous hepatitis B ge antigen (HBeAg) seroconversion than genotype B patients, stored serum samples from 146 Taiwanese adult HBeAg-positive hepatitis B carriers followed-up for a mean of 52 months (range, 12-120 months) were tested for HBV genotype by a molecular method. Genotype C patients were significantly older than genotype B patients (mean age, 37 +/- 12 vs. 29 +/- 10 years, P < 0.001). During the follow up period, genotype C patients had a significantly lower rate of spontaneous HBeAg seroconversion than genotype B patients (27 vs. 47%, P < 0.025). Spontaneous HBeAg seroconversion occurred one decade later in genotype C patients compared with genotype B patients. Multivariate analyses identified age < or =35 years (odds ratio: 2.08; 95% confidence interval [CI], 1.07-4.0; P < 0.05), high baseline serum alanine aminotransferase level (odds ratio: 2.34; 95%CI, 1.39 4.09; P < 0.005), and HBV genotype B (odds ratio: 1.94; 95%CI, 1.03-3.63; P < 0.05) as independent factors associated with spontaneous HBeAg seroconversion. In conclusion, genotype C patients, compared to genotype B patients, have a delayed HBeAg seroconversion in the immune clearance phase of chronic HBV infection, which may contribute to a more progressive liver disease and more refractory to antiviral therapy. PMID- 14748060 TI - Direct evidence that naturally occurring mutations within hepatitis B core epitope alter CD4+ T-cell reactivity. AB - Exacerbations of chronic hepatitis B have been associated with accumulation of mutations in the HBV core gene, with amino acid (aa) substitutions clustering between aa 50 and 69. This region of the nucleocapsid protein is known as an immunodominant epitope for CD4+ T-lymphocytes, however the impact of these mutations on T-cell reactivity has not been investigated. For this purpose, we undertook fine mapping of the reactivity of peripheral blood lymphocytes, isolated from patients with acute (n = 8) or chronic hepatitis B (n = 10), against a panel of branched synthetic peptides. The peptide aa sequences corresponded to the wild type HBV (aa 50-69), or contained 1-3 aa changes derived on the basis of naturally occurring mutations. In four of eight patients with acute hepatitis B the wild type peptide 50-69, which corresponded to the core gene sequence of HBV present in these patients, induced a strong T-cell proliferative response. In the same cases, the T-cell response to the mutant peptides was altered at various degrees, depending on the number and the position of aa changes. The most pronounced inhibition of CD4+ T-cell response (between 44 and 92%) was caused by a peptide ligand with two aa substitutions at positions 64 and 67. These results demonstrate that mutations within immunodominant epitopes of the HBV nucleocapsid can affect the CD4+ T-lymphocyte reactivity, which may have a role for the accumulation of certain HBV strains after hepatitis flares during the course of chronic HBV infection. PMID- 14748061 TI - Clade analysis and surface antigen polymorphism of hepatitis B virus American genotypes. AB - Eight genotypes (A-H) of hepatitis B virus (HBV) have been described, HBV genotypes F and H being autochthonous to America. HBV genotype F has been classified in four clusters. The objective of this study was to gain insight into the molecular epidemiology of HBV American genotypes, as well as to analyze the genotype-related polymorphism in some functional domains of the surface proteins. The sequences of the S region of 106 isolates genotype F and H were analyzed, out of which 47 isolates genotype F circulated in different Venezuelan populations. Most of the Venezuelan isolates genotype F were grouped in cluster III (n = 39) and 7 in cluster II. One isolate obtained from a blood donor could not be classified in any clade and harbored amino acid substitutions characteristic of a vaccine escape mutant (G145R) and a stop codon in the surface antigen. Amino acid analysis of the PreS and S gene products showed unique genetic characteristics in genotype F and H sequences in some important domains involved in the early steps of infection. Out of 30 available sequences, two complete genome sequences of HBV genotype F from Venezuela were obtained. Phylogenetic analysis of these complete genomes confirmed the presence of four clusters inside genotype F, differing in more than 4% nucleotide divergence. Our extended analysis showed that genotype F clades Ia, III, and IV exhibit a restricted geographic distribution (Central America, the North and the South of South America, respectively) while clades Ib and II are found in all the Americas except in the Northern South America and North America respectively. PMID- 14748062 TI - Characterization of mimotopes mimicking an immunodominant conformational epitope on the hepatitis C virus NS3 helicase. AB - The hepatitis C virus (HCV) nonstructural 3 (NS3) protein is composed of an amino terminal protease and a carboxyl terminal RNA helicase. NS3 contains major antigenic epitopes. The antibody response to NS3 appears early in the course of infection and is focused on the helicase region. However, this response cannot be defined by short synthetic peptides indicating the recognition of conformation dependent epitopes. In this study, we have screened a dodecapeptide library displayed on phage with anti-NS3 mouse monoclonal antibodies (mAbs) that compete with each other and human anti-HCV NS3 positive sera. Two peptides (mimotopes) were selected that appeared to mimic an immunodominant epitope since they were recognized specifically by the different anti-NS3 mAbs of the study and by human sera from HCV infected patients. Homology search between the two mimotopes and the NS3 sequence showed that one of the two peptides shared amino acid similarities with NS3 at residues 1396-1398 on a very accessible loop as visualized on the three-dimensional structure of the helicase domain whereas the other one had two amino acids similar to nearby residues 1376 and 1378. Reproduced as synthetic dodecapeptides, the two mimotopes were recognized specifically by 19 and 22, respectively, out of 49 sera from HCV infected patients. These mimotopes allowed also the detection of anti-NS3 antibodies in sera of HCV patients at the seroconversion stage. These results suggest that the two NS3 mimotopes are potential tools for the diagnosis of HCV infection. PMID- 14748063 TI - Lack of clinical significance of variability in the internal ribosome entry site of hepatitis C virus. AB - The extreme 5'-proximal sequence of the hepatitis C virus (HCV) genome including the 5' non-coding region (5'NCR) of 341 nucleotide long and the first 30 nucleotides of the core region is highly conserved among different HCV genotypes. It contains a segment termed Internal Ribosome Entry Site (IRES) that regulates the cap-independent translation of HCV-RNA to polyprotein. Sequence variability in this region has important implications for structural organisation and function of the IRES element and could correlate with HCV RNA concentration or response to antiviral therapy. Fourteen patients (seven women, seven men) with chronic hepatitis C were separated into two groups according to their response to antiviral therapy. Seven of these were sustained responders to treatment by Interferon alpha 2b and Ribavirin and seven were non-responders. After cloning sequencing, the IRES (nt 21 to 374) appears to be structurally highly conserved. However some variability was found between the different isolates obtained: 209 substitutions with a median of four variants/patients. Comparison of the number of variants present in the viral population of the sustained responders and non responders patients do not show any difference. Positioning of the mutations on the predicted IRES secondary structure showed that the distribution of the mutations and their apparition frequency were different between the two groups. The translation initiator AUG-4 codon, located in the stem-loop IV, is never modified. Variations observed in the IRES are not a parameter of response to antiviral therapy, but the integrity of this region is a necessary condition to maintain its activity. PMID- 14748064 TI - Kinetics of viremia and acute liver injury in relation to outcome of neonatal woodchuck hepatitis virus infection. AB - The kinetics of serum viral responses and acute liver injury were studied during neonatal woodchuck hepatitis virus (WHV) infection in relation to the chronic or resolved outcome. The mean concentrations of serum WHV DNA and surface antigen were significantly higher by week 10 post infection in chronic infections compared to resolving infections, and diverged even further by the time of peak viral load development in serum (week 12). After week 12, these viral markers were detected less frequently with time and at lower concentrations in the resolved outcome. In both outcomes, mean serum activities of hepatic enzymes became increased significantly above baseline by weeks 10-12, peaked at week 14, and normalized by weeks 20-22, thus indicating transient acute liver injury. The increasing liver injury responses were comparable between outcomes at week 12, when serum viral load was markedly higher in the developing chronic infections. This suggested a deficiency in early non-cytolytic control of infection in the chronic outcome. At week 14, liver injury was significantly greater in the resolved outcome and associated with higher mean Fas ligand (FasL) and perforin messenger RNAs (mRNAs) in liver compared to the chronic outcome. This indicated greater immune-mediated killing of infected hepatocytes during resolution. Thus, chronicity as an outcome of neonatal WHV infection develops relatively early during the acute phase of infection due to reduced immune-mediated clearance of infected hepatocytes by both cytolytic and non-cytolytic processes. PMID- 14748065 TI - Measurement of antirotavirus IgM/IgA/IgG responses in the serum samples of Indian children following rotavirus diarrhoea and their mothers. AB - Rotavirus specific, serum IgM/IgA/IgG levels among hospitalized children and their respective mothers were determined. Children were grouped as having rotavirus diarrhoea (RVD) and non-rotavirus diarrhoea (NRVD) on the basis of fecal excretion measured by ELISA and RT-PCR. Although IgM seropositivity was observed among children of both the groups, it was significantly higher in the acute as well as convalescent phase serum samples (P < 0.05 for both) of RVD group. Five out of ten acute sera from the NRVD group were positive for IgM and seven showed IgA/IgG seroconversion indicating rotavirus infection among these children in the past. It was noted that, three out of 24 mothers' sera from RVD group, showed presence of IgM in the serum collected during convalescence of their children. The observation suggests, subclinical rotavirus infection among mothers probably contacted from their children. This is supported by the seroconversion for IgA/IgG among these three mothers. Such a phenomenon was not noticed among the mothers from NRVD group. In general, IgA positivity did not vary significantly among the children from both the groups. IgA seropositivity was significantly higher (P < 0.001) from children of RVD group as compared to healthy group of children following rotavirus infection. From RVD group, all the child patients and 12 mothers out of 24 (50%) showed IgA/IgG seroconversion. None of the mothers from NRVD group showed seroconversion. Serum samples of healthy children and adults, showed IgM positivity at equal level (10%), but a significant difference (P < 0.01) was observed in IgA positivity. In conclusion, subclinical transmission of rotavirus infection from children to their mothers may occur. Seroconversion alone cannot be considered as a marker of rotavirus diarrhoea in children. Moreover, about 40-50% of subjects lacked rotavirus specific IgA at protective levels, making them susceptible to rotavirus infection. PMID- 14748067 TI - Apoptosis in dengue virus infected liver cell lines HepG2 and Hep3B. AB - While both in vivo and in vitro evidence has suggested that liver cells undergo apoptosis in response to dengue virus infection, little is known about the mechanism of induction. Given that the p53 tumour suppressor gene is a key mediator of apoptosis, we sought to define the role of this gene in response to dengue virus infection. After infection, a p53 wild type liver cell line (HepG2) showed changes consistent with apoptosis including alterations of cell morphology, cellular detachment and DNA laddering. However, p53 was neither up regulated, nor showed any evidence of complexing with dengue virus proteins as determined by immunoprecipitation. Infection of a p53 null liver cell line (Hep3B) also produced changes consistent with the induction of apoptosis. While the profile of the cells undergoing apoptosis in each cell line was similar as determined by flow cytometry, the absolute levels were markedly different with up to 90% of Hep3B cells undergoing apoptosis compared to only 20% of HepG2 cells at day 5 post infection. By day 7, all Hep3B infected cells were dead. In contrast, it proved possible to culture dengue virus infected HepG2 cells for 3 months. Viral progeny released from the p53 null cell line were nine-fold higher per attached cell than from the p53 wild type cell line. These results suggest that, while induction of apoptosis in liver cells is mediated by a non-p53 regulated pathway, p53 may play a role in restricting the level of viral progeny to below a critical level at which apoptosis is triggered. PMID- 14748066 TI - Mucosal arenavirus infection of primates can protect them from lethal hemorrhagic fever. AB - Arenaviruses are transmitted from rodents to human beings by blood or mucosal exposure. The most devastating arenavirus in terms of human disease is Lassa fever virus, causing up to 300,000 annual infections in West Africa. We used a model for Lassa fever in which Rhesus macaques were infected with a related virus, lymphocytic choriomeningitis virus (LCMV). Our goals were to determine the outcome of infection after mucosal inoculation and later lethal challenge, to characterize protective immune responses, and to test cross-protection between a virulent (LCMV-WE) and an avirulent (LCMV-ARM) strain of virus. Although intravenous infections in the monkey model were uniformly lethal, intragastric infections recapitulated the spectrum of clinical outcomes seen in human exposure to Lassa fever virus: death, recovery from disease, and most often, subclinical infection. Plaque neutralization, ELISA, lymphocyte proliferation, and chromium release assays were used to monitor humoral and cellular immune responses. Cross protection between the two strains was observed. The three out of seven monkeys that experienced protection were also the three with the strongest cell-mediated immunity. PMID- 14748068 TI - Elevation of soluble VCAM-1 plasma levels in children with acute dengue virus infection of varying severity. AB - Approximately 1,000 million infections with dengue viruses are estimated to occur annually. The majority of the cases develop mild disease, whereas only small proportion of the infected individuals develop severe hemorrhagic manifestations at the end of the acute phase of illness. In this study, the value of plasma levels of vascular cell adhesion molecule 1 (VCAM-1) in the pathogenesis and prognosis of dengue illness was investigated in children with dengue infections of varying severity. The plasma levels of soluble VCAM-1 (sVCAM-1) were measured in serial plasma samples obtained from 168 children aged between 7 months and 14 years with confirmed dengue infection. Of those children, 71 were suffering from dengue fever, 30 were suffering from dengue hemorrhagic fever, and 67 were suffering from dengue shock syndrome. Plasma samples obtained from 21 patients with febrile illness other than dengue served as controls. A commercially available kit (R&D Systems, Oxon, UK) was used to measure the levels of sVCAM-1 in plasma samples. sVCAM-1 was elevated during acute dengue infection, and significantly elevated among dengue shock syndrome patients as compared to dengue fever or dengue hemorrhagic fever patients (P < 0.05). Statistical analysis revealed that sVCAM-1 was associated with dengue disease severity and the time post infection (acute vs. convalescent phase) and not with age, sex, or previous exposure of the patients to dengue infection. A significant difference was found in the plasma levels of sVCAM-1 between dengue shock syndrome and dengue fever patients, however, the prognostic value of this marker in the acute stage of dengue illness proved to be limited. These data also favor to study the further elucidation of the role of sVCAM-1 in the pathogenesis of dengue infections. PMID- 14748069 TI - Mechanisms of beta cell death during restricted and unrestricted enterovirus infection. AB - Coxsackie B virus (CVB-5) infections potentially trigger and accelerate pancreatic beta cell damage leading to type 1 diabetes. In vivo, all viruses face natural resistance mediated by various host factors which restrict the progression of infection. Thus, the aims of this study were to generate a tissue culture model of restricted coxsackie B virus infection in primary islet cells by preventing the production of viral progeny with a selective inhibitor of viral RNA replication and to investigate the mechanisms of virus-induced islet cell death during productive and restricted infective conditions. Cultured foetal porcine islet cells were infected effectively with the prototype strain of coxsackievirus B5 (CVB-5). Nuclear viability stainings and electron microscopy showed productive infection to result in dominantly necrotic cell death with additional slight induction of apoptosis during the 7 days of follow-up. The restricted conditions were created by addition of guanidine-hydrochloride (G-HCl) into culture medium. At 1 mM concentration, it significantly protected the infected cells from necrosis and thus maintained high viability. This was associated with increased significantly apoptosis. In perifusion analysis, the cellular ability to release insulin was reduced, although the metabolic integrity was preserved as shown by MTT-analysis and cellular ATP levels. These data show that restriction of CVB-5 replication with G-HCl protects islet cells against virus-induced necrosis. However, restriction of viral replication shifts the mechanism of cell death from necrosis toward apoptosis. A slowly progressing subclinical infection of islets could thus lead to increased beta-cell apoptosis. PMID- 14748070 TI - Human herpesvirus-8 infection in pregnancy and labor: lack of evidence of vertical transmission. AB - To investigate whether vertical transmission of the human herpesvirus 8 (HHV-8) may occur during pregnancy or at delivery, we enrolled 295 women recruited attending the Division of Obstetrics and Gynecology of a University Teaching of Rome Tor Vergata, S. Eugenio Hospital. The study population was divided in two groups: 245 pregnant women who underwent amniocentesis for genetic screening at 16-18 weeks gestation (group 1) and 50 women at the childbirth (group 2). Maternal blood was obtained from all women. Amniotic fluid (group 1) and cord blood (group 2) were obtained at midtrimester and at delivery, respectively. The presence of anti-HHV-8 antibodies in serum samples was investigated by an immunfluorescence assay. All amniotic fluids, maternal blood, and cord blood samples from HHV-8 seropositive women were tested for the presence of HHV-8 DNA sequences by the polymerase chain reaction. Thirty women, 27 of the group 1 and three of the group 2, were found to have anti-HHV-8 antibodies. Two neonates of the three seropositive mothers of the group 2 had anti-HHV-8 antibodies in cord blood. HHV-8 DNA sequences were detected in the blood of one woman of the group 2. None of the amniotic fluid and cord blood samples had detectable HHV-8 DNA sequences. This study suggests that vertical transmission of HHV-8 is unlikely or, at least, very rare. PMID- 14748071 TI - Simultaneous detection and differentiation of human polyomaviruses JC and BK by a rapid and sensitive PCR-ELAHA assay and a survey of the JCV subtypes within an Australian population. AB - Human polyomaviruses JCV and BKV can cause several clinical manifestations in immunocompromised hosts, including progressive multifocal leukoencephalopathy (PML) and haemorrhagic cystitis. Molecular detection by polymerase chain reaction (PCR) is recognised as a sensitive and specific method for detecting human polyomaviruses in clinical samples. In this study, we developed a PCR assay using a single primer pair to amplify a segment of the VP1 gene of JCV and BKV. An enzyme linked amplicon hybridisation assay (ELAHA) using species-specific biotinylated oligonucleotide probes was used to differentiate between JCV and BKV. This assay (VP1-PCR-ELAHA) was evaluated and compared to a PCR assay targeting the human polyomavirus T antigen gene (pol-PCR). DNA sequencing was used to confirm the polyomavirus species identified by the VP1-PCR-ELAHA and to determine the subtype of each JCV isolate. A total of 297 urine specimens were tested and human polyomavirus was detected in 105 specimens (35.4%) by both PCR assays. The differentiation of JCV and BKV by the VP1-PCR-ELAHA showed good agreement with the results of DNA sequencing. Further, DNA sequencing of the JCV positive specimens showed the most prevalent JCV subtype in our cohort was 2a (27%) followed by 1b (20%), 1a (15%), 2c (14%), 4 (14%) and 2b (10%). The results of this study show that the VP1-PCR-ELAHA is a sensitive, specific and rapid method for detecting and differentiating human polyomaviruses JC and BK and is highly suitable for routine use in the clinical laboratory. PMID- 14748072 TI - Latent Human Papillomavirus infection is comparable in the larynx and trachea. AB - Recurrent respiratory papillomas are benign airway tumors caused by Human Papillomaviruses (HPVs) types 6 and 11. The disease is characterized by multiple recurrences of papillomas following surgical removal, caused by activation of latent HPV DNA. Most patients have laryngeal disease, while only a small subset has tracheal involvement. We have asked whether the lower frequency of tracheal papillomas was due to reduced prevalence of latent/subclinical tracheal HPV infection or reduced likelihood of activation to clinically apparent disease. A total of 121 biopsies of clinically normal laryngeal and tracheal tissues from 61 patients with laryngeal papillomas were analyzed for HPV DNA by polymerase chain reaction, confirmed by Southern blot hybridization. Patients were followed for 3 18 years (mean = 5.5 +/- 4.4), with only one developing subsequent tracheal disease. There was no significant difference in prevalence of latent HPV DNA between larynx and trachea, analyzing either those patients with a single biopsy or those with more than one biopsy of larynx, trachea, or both. There was also no significant difference between tracheal latency with HPV 6 and HPV 11. We conclude that HPV infects tracheal mucosa and is maintained as a latent infection in the trachea as efficiently as in the larynx. Therefore, we propose that the low frequency of tracheal disease reflects a lower frequency of HPV activation, and postulate that cellular factors that differ between the stratified squamous epithelium of the larynx and the ciliated pseudo-stratified columnar epithelium of the trachea contribute to this difference. PMID- 14748073 TI - Infection with Human Papillomavirus alters expression of the small proline rich proteins 2 and 3. AB - Human papillomavirus (HPV) does not induce lysis of infected keratinocytes, and the exact mechanisms of viral escape are not known. As keratinocytes differentiate, the cornified cell envelope (CCE) develops, providing a protective barrier to the host. Our prior studies have identified abnormalities in CCEs isolated from genital epithelium infected with HPV 11 (a low-risk HPV type) and HPV 59 (a high-risk HPV type). These abnormalities included reduced thickness and increased fragility compared to CCEs in healthy epithelium. Transcription of loricrin is also reduced in HPV 11- and 59-infected epithelium. In this study, uninfected and HPV 11- or 59-infected human genital epithelium were examined for expression of the small proline rich proteins (SPRs), which serve as cross linking proteins within the CCE. Limiting cycle RT-PCR was performed to detect the various SPR transcripts in HPV 11- and 59-infected, or uninfected epithelium. Immunohistochemical analysis and immunoblot assays were performed to analyze the distribution and quantity of SPR2A, SPR2B, and SPR3. SPR2B transcripts were moderately increased in the HPV 11- and 59-infected tissues and SPR3 transcripts were significantly increased in HPV 11-infected tissues and minimally increased in HPV 59-infected tissues. SPR2B protein quantities were moderately increased while SPR2A was not significantly changed. SPR3 protein, while not present in uninfected epithelium, was detected in abundance in HPV 11-infected tissue. We conclude that low-risk and high-risk HPVs share the ability to alter expression of CCE proteins, although the exact mechanisms may differ. Expression of individual SPRs differed between these types and these alterations may play a role in fragility of CCEs in HPV infection. PMID- 14748074 TI - Simultaneous detection of fourteen respiratory viruses in clinical specimens by two multiplex reverse transcription nested-PCR assays. AB - There is a need for rapid, sensitive, and accurate diagnosis of lower respiratory tract infections in children, elderly, and immunocompromised patients, who are susceptible to serious complications. The multiplex RT-nested PCR assay has been used widely for simultaneous detection of non-related viruses involved in infectious diseases because of its high specificity and sensitivity. A new multiplex RT-PCR assay is described in this report. This approach includes nested primer sets targeted to conserve regions of human parainfluenza virus haemagglutinin, human coronavirus spike protein, and human enterovirus and rhinovirus polyprotein genes. It permits rapid, sensitive, and simultaneous detection and typing of the four types of parainfluenza viruses (1, 2, 3, 4AB), human coronavirus 229E and OC43, and the generic detection of enteroviruses and rhinoviruses. The testing of 201 clinical specimens with this multiplex assay along with other one formerly described by our group to simultaneously detect and type the influenza viruses, respiratory syncytial viruses, and a generic detection of all serotypes of adenovirus, covers the detection of most viruses causing respiratory infectious disease in humans. The results obtained were compared with conventional viral culture, immunofluorescence assay, and a third multiplex RT-PCR assay for all human parainfluenza viruses types described previously. In conclusion, both multiplex RT-PCR assays provide a system capable of detecting and identifying simultaneously 14 different respiratory viruses in clinical specimens with high sensitivity and specificity, being useful for routine diagnosis and survey of these viruses within the population. PMID- 14748075 TI - Increased detection of rotavirus using a real time reverse transcription polymerase chain reaction (RT-PCR) assay in stool specimens from children with diarrhea. AB - Six-hundred and twenty-six stool specimens collected from children with diarrhea over a 12-month period were tested for rotavirus using a real time reverse transcription-polymerase chain reaction (RT-PCR) assay, a conventional nested PCR assay and by electron microscopy (EM). A fragment of 87 bp in a highly-conserved region of non-structural protein 3 (NSP3) in rotavirus genome was amplified by a single-step RT-PCR protocol in a closed-tube system. Rotavirus was detected in 123 samples (20%) with the real time RT-PCR assay, 113 samples (18%) with the nested-PCR assay, and 79 samples (13%) with EM. Using serial diluted nucleic acid extract, we compared the sensitivity of real time RT-PCR with conventional RT-PCR and conventional nested PCR assays. Real time RT-PCR was two to four logs more sensitive than the conventional assays. The reaction time required for the RT-PCR assay is about half the time required for the conventional nested-PCR. The real time RT-PCR assay is both simple and rapid with advantages including enhanced sensitivity and a lower risk for cross-contamination making it a useful tool for the detection of rotavirus in various situations including sporadic gastroenteritis, outbreaks, and environmental investigations. G(1) was the predominant type (89%), followed by G(2) (10%), and G(4) (1%). No rotavirus of G(3), G(8), and G(9) types were found. The peak season for rotavirus infection was January to May in northern Alberta. PMID- 14748076 TI - Detection and genotyping of TT virus in healthy and subjects with HBV or HCV in different populations in the United Arab Emirates. AB - TT virus (TTV) and TTV-like viruses (TTVLs) have been reported to be associated with non-A-E hepatitis. To determine the rate of infection and genotypic characteristics of TTV in the United Arab Emirates (UAE), a total of 449 serum samples representing different populations in the UAE and comprising healthy as well as patients positive for HBsAg and HCV were screened. National subjects (n = 200) and non-nationals residing in the UAE (n = 249) were tested by PCR. The results obtained showed that the rate of TTV infection in healthy nationals, and those with HBsAg or antibody to HCV were 34.9, 97.9, and 95.7, respectively, compared to 89.1% (115/129), 89.2% (66/74), and 84.8% (39/46), respectively, in non-nationals. Sequence analysis of the untranslated region (UTR) using 71 clones generated from the PCR products of eight serum samples from healthy individuals (four nationals and four non-nationals) showed that 83.1% of the TTV clones were classified into groups 1-4, whereas 16.9% into possibly new genotype(s). The analysis also revealed that healthy national subjects carried multiple viruses. Phylogenetic analysis of representative sequences revealed clustering of clones into at least five major groups. Also, when compared to reference genotypes (from GenBank), two of our clones belonged to two previously identified genotypes. Non significant gender differences were seen in all ethnic groups studied (P > 0.05). In conclusion, the rate of TTV infection in the UAE nationals is significantly lower (P < 0.05) than that of the non-nationals and several genotypes were isolated with common multi-infections. PMID- 14748078 TI - The life and death of sponge cells. AB - Cell viability is an essential touchstone in the study of the effect of medium components on cell physiology. We developed a flow-cytometric assay to determine sponge-cell viability, based on the combined use of fluorescein diacetate (FDA) and propidium iodide (PI). Cell fluorescence measurements based on incubation of cells with FDA or PI resulted in a useful and reproducible estimate of the viability of primary sponge-cell cultures. We studied the effects of temperature, ammonium, and the fungicide amphotericin B on the viability of a primary-cell culture from the marine sponge Suberites domuncula using the aforementioned flow cytometric assay. S. domuncula cells die rapidly at a temperature of >or=22 degrees C, but they are insensitive to ammonium concentrations of up to 25 mM. Amphotericin B, which is frequently used in sponge-cell culture media, was found to be toxic to S. domuncula cells. PMID- 14748079 TI - Transglutaminase-mediated modification of glutamine and lysine residues in native bovine beta-lactoglobulin. AB - Bovine beta-lactoglobulin (BLG) is a major component in whey and its physical properties are important for the texture of many dairy-based foods. Modification of proteins with transglutaminase from Streptoverticillium mobaraense (MTGase) can be used to alter their physical properties. MTGase-mediated modification of native BLG was until now, however, not effective. Here we report a method that allows for the enzymatic modification of native BLG with MTGase. Lysines 8, 77, and 141 were modified with alpha-N-carbobenzyloxy-glutamine-glycine and glutamines 35, 59, 68,and 155 were modified with 6-aminohexanoic acid under nonreducing and nondenaturing conditions. MTGase-mediated BLG crosslinking is hampered by the low reactivity of the lysines and enzymatic deamidation of the glutamines prevails. Modification of BLG with poly-lysine yields a BLG derivative with increased affinity for the water-air interface and stronger surface tension lowering capacities than normal BLG. Hence, this modification method offers the opportunity to change the functional properties of BLG and to prepare novel protein foods. PMID- 14748080 TI - New tools for mass isotopomer data evaluation in (13)C flux analysis: mass isotope correction, data consistency checking, and precursor relationships. AB - 13C metabolic flux analysis (MFA) is based on carbon-labeling experiments where a specifically (13)C labeled substrate is fed. The labeled carbon atoms distribute over the metabolic network and the label enrichment of certain metabolic pools is measured by using different methods. Recently, MS methods have been dramatically improved-large and precise datasets are now available. MS data has to be preprocessed and corrected for natural stable mass isotopes. In this article we present (1). a new elegant method to correct MS measurement data for natural stable mass isotopes by infinite dimensional matrix calculus and (2). we statistically analyze and discuss a reconstruction of labeling pattern in metabolic precursors from biosynthesis molecules. Moreover, we establish a new method for consistency checking of MS spectra that can be applied for automatic error recognition in high-throughput flux analysis procedures. Preprocessing the measurement data changes their statistical properties which have to be considered in the subsequent parameter fitting process for (13)C MFA. We show that correcting for stable mass isotopes leads to rather small correlations. On the other hand, a direct reconstruction of a precursor labeling pattern from an aromatic amino acid measurement turns out to be critical. Reasonable results are only obtained if additional, independent information about the labeling of at least one precursor is available. A versatile MatLab tool for the rapid correction and consistency checking of MS spectra is presented. Practical examples for the described methods are also given. PMID- 14748081 TI - Genome-wide transcriptional response of a Saccharomyces cerevisiae strain with an altered redox metabolism. AB - The genome-wide transcriptional response of a Saccharomyces cerevisiae strain deleted in GDH1 that encodes a NADP(+)-dependent glutamate dehydrogenase was compared to a wild-type strain under anaerobic steady-state conditions. The GDH1 deleted strain has a significantly reduced NADPH requirement, and therefore, an altered redox metabolism. Identification of genes with significantly changed expression using a t-test and a Bonferroni correction yielded only 16 transcripts when accepting two false-positives, and 7 of these were Open Reading Frames (ORFs) with unknown function. Among the 16 transcripts the only one with a direct link to redox metabolism was GND1, encoding phosphogluconate dehydrogenase. To extract additional information we analyzed the transcription data for a gene subset consisting of all known genes encoding metabolic enzymes that use NAD(+) or NADP(+). The subset was analyzed for genes with significantly changed expression again with a t-test and correction for multiple testing. This approach was found to enrich the analysis since GND1, ZWF1 and ALD6, encoding the most important enzymes for regeneration of NADPH under anaerobic conditions, were down regulated together with eight other genes encoding NADP(H)-dependent enzymes. This indicates a possible common redox-dependent regulation of these genes. Furthermore, we showed that it might be necessary to analyze the expression of a subset of genes to extract all available information from global transcription analysis. PMID- 14748082 TI - Poly-3-hexyl thiophene Langmuir-Blodgett films for application to glucose biosensor. AB - Langmuir-Blodgett films of poly(3-hexyl thiophene) have been prepared by simultaneous entrapment of glucose oxidase and transferred onto the indium-tin oxide coated glass plates. The films have been characterized by FTIR spectroscopy and detailed response studies have been performed with respect to glucose concentration, temperature, and storage time. These P3HT/SA/GOX electrodes have been utilized for glucose estimation from 100-500 mg/dL by amperometric method. The electrodes have been found to have sensitivity of 0.75 nA/mg/dL detection limit of 50 mg/dL and shelf life of about 75 days at 4 degrees C. PMID- 14748083 TI - Exogenous cadherin microdisplay can interfere with endogenous signaling and reprogram gene expression in cultured hepatocytes. AB - We recently found that the basal micro substrate presentation of E-cadherin, a key cell-cell adhesion molecule in the liver, can modulate hepatocellular proliferative potential and differentiated function (Brieva and Moghe, in press). In the current study, we established a similar experimental model involving rat hepatocytes cultured on collagen and incorporated 5 microm polystyrene microbeads functionalized with Protein A-anchored E-cadherin/human lgG Fc chimeric fusion constructs. We investigated the cadherin governed dose-response of cell proliferative potential and quantified the underlying changes in intracellular gene signaling processes. Hepatocellular proliferative potential was found to be intensified with an increase in the microdisplay of acellular cadherins and this effect was offset by increased cell seeding density. Notably, we report that following overnight exposure to acellular cadherins, the expression of genes known to mediate the control of cell proliferation, cyclin D1 and c-myc, was upregulated, while the expression of differentiation-related genes, namely albumin and cytochrome p450 II B1, was reduced. The exposure of cell cultures to exogenous cadherins was found to markedly disrupt the localization of endogenous E-cadherin and beta-catenin to junctions at cell-cell contacts and cause a quantitative decrease in the endogenous cadherin protein levels. Based on all of our observations, we propose that the acellular presentation of E-cadherin chimeras competitively disrupts endogenous cadherin containing complexes at cell cell junctions and increases intracellular cadherin turnover, thereby promoting beta-catenin mediated signaling, which ultimately engenders an increase in cell proliferative potential and a decrease in differentiated function. PMID- 14748084 TI - Affinity purification of plasmid DNA by temperature-triggered precipitation. AB - This report describes a new plasmid DNA purification method, which takes advantage of the DNA-binding affinity and specificity of the bacterial metalloregulatory protein MerR, and of the temperature responsiveness of elastin like proteins (ELPs). Upon increasing the temperature, ELP undergoes a reversible phase transition from water-soluble forms into aggregates, and this property was exploited for the precipitation of plasmid DNA containing the MerR recognition sequence by a simple temperature trigger. In one purification step, plasmid DNA was purified from E. coli cell lysates to a better purity than that prepared by a standard alkaline purification method, with no contaminating chromosomal DNA and cellular proteins. This protein-based approach, in combination with the reversible phase transition feature of ELP, makes the outlined method a promising candidate for large-scale purification of plasmid DNA for sensitive applications such as nonviral gene therapy or DNA vaccines. PMID- 14748085 TI - On-line monitoring of important organoleptic methyl-branched aldehydes during batch fermentation of starter culture Staphylococcus xylosus reveal new insight into their production in a model fermentation. AB - A small fermentor (55 mL) was directly interfaced to a membrane inlet mass spectrometer for continuous on-line monitoring of oxygen and volatile metabolites during batch fermentations of the starter culture Staphylococcus xylosus. Using this technique, we were able to correlate production of the very important flavor compounds 2-methylbutanal, 3-methylbutanal, and 2-methylpropanal with various growth conditions. We found that the aldehydes were present in the culture broth only as transient metabolites. They were produced in the exponential growth phase, reached a maximum concentration when the culture became anaerobic, and then they rapidly disappeared from the culture medium. This general pattern was observed for three different strains of S. xylosus and S. carnosus. Small amounts of inoculum or increased exposure to oxygen were found to favor production of the aldehydes as a result of a longer aerobic growth period. Growing S. xylosus under conditions resembling those in a fermented sausage revealed that NaCl (5%) increased aldehyde production considerably, whereas KNO(3) (0.03%) or NaNO(2) (0.03%) had little effect. A lowering of pH from 7.2 to 6.0 reduced cell density, but had a minor affect on aldehyde production. PMID- 14748086 TI - Detection, isolation, and characterization of exopolysaccharide produced by a strain of Phormidium 94a isolated from an arid zone of Mexico. AB - Phormidium 94a, a cyanobacteria that produces extracellular polymeric substances (EPS), was isolated from arid soils of Mexico. Microscopic localization, using histochemical techniques like the Toluidine blue technique, was done in order to demonstrate the presence of EPS. Acetone was added to precipitate the EPS. In this study we characterized the EPS by GC, HPLC, and IR techniques. The highest fraction of EPS had a molecular weight of 2000 kDa. The sugar composition was galactose, mannose, galacturonic acid, arabinose, and ribose in the three main fractions, and the sugar ratio found was different in each fraction. The low EPS concentrations had a Newtonian behavior, when the concentrations were increased, the behavior changed to pseudoplastic. The EPS rheulogical behavior is similar to low viscosity arabic gum. Also, it was found that an increase in viscosity occurred at longer hydration time. More rheological and toxicological studies are required in order to analyze its possible application in food industries. PMID- 14748087 TI - Resistance to signal activation governs design features of the MAP kinase signaling module. AB - Given its broad influence over numerous cell functions, redesigning the mitogen activated protein (MAP) kinase signaling module would offer a powerful means to engineer cell behavior. Early challenges include identifying quantitative module features most relevant to biological function and developing simple design rules to predictably modify these features. This computational study delineates how features such as signal amplification, input potency, and dynamic range of output may be tuned by manipulating chief module components. Importantly, the model construction identifies a metric of resistance to signal activation that quantitatively predicts module features and design trade-offs for broad perturbations in kinase and phosphatase expression. Its predictive utility extends to dynamic properties such as signal lifetime, which often dictates MAP kinase effect on cell function. Taken together, we propose that predictably altering MAP kinase signaling by tuning resistance is not only a feasible engineering strategy, but also one exploited by natural systems to allow each MAP kinase to exert pleiotropic effects in a context-dependent manner. External stimuli not only activate kinases, but also alter phosphatase expression and activity, thereby reconfiguring a single module for quantitatively distinct modes of signaling such as transient vs. sustained dynamics, each with unique effects on cell function. PMID- 14748088 TI - Penicillin acylase-catalyzed synthesis of beta-lactam antibiotics in highly condensed aqueous systems: beneficial impact of kinetic substrate supersaturation. AB - Advantages of performing penicillin acylase-catalyzed synthesis of new penicillins and cephalosporins by enzymatic acyl transfer to the beta-lactam antibiotic nuclei in the supersaturated solutions of substrates have been demonstrated. It has been shown that the effective nucleophile reactivity of 6 aminopenicillanic (6-APA) and 7-aminodesacetoxycephalosporanic (7-ADCA) acids in their supersaturated solutions continue to grow proportionally to the nucleophile concentration. As a result, synthesis/hydrolysis ratio in the enzymatic synthesis can be significantly (up to three times) increased due to the nucleophile supersaturation. In the antibiotic nuclei conversion to the target antibiotic the remarkable improvement (up to 14%) has been gained. Methods of obtaining relatively stable supersaturated solutions of 6-APA, 7-ADCA, and D-p hydroxyphenylglycine amide (D-HPGA) have been developed and syntheses of ampicillin, amoxicillin, and cephalexin starting from the supersaturated homogeneous solutions of substrates were performed. Higher synthetic efficiency and increased productivity of these reactions compared to the heterogeneous "aqueous solution-precipitate" systems were observed. The suggested approach seems to be an effective solution for the aqueous synthesis of the most widely requested beta-lactam antibiotics (i.e., amoxicillin, cephalexin, cephadroxil, cephaclor, etc.). PMID- 14748089 TI - Expression of recombinant vitellogenin in the yeast Pichia pastoris. AB - Vitellogenin (Vtg) plays vital roles as precursor to the yolk proteins and as carrier for lipids, carbohydrates, phosphates, metal ions, vitamins, and hormones into the oocytes during the massive deposition of yolk nutrients for subsequent nourishment of the developing embryos. Reproductive success is highly sensitive to the nutritional quality of the broodstock diet, which greatly affects the egg and larval viability. We present a novel strategy for genetically engineering a Pichia pastoris yeast strain that constitutively produces recombinant Vtg (rVtg), for application as an enriched feed. The tilapia Oreochromis aureus Vtg (OaVtg) cDNA (5.3 kb) was cloned into a nonsecretory pGAPZA vector. Clones containing up to 31 copies of glyceraldehyde-3-phosphate dehydrogenase (GAP)-promoter-driven Vtg expression cassettes were isolated. These clones expressed a membrane associated intracellular rVtg protein of 194 kDa, constituting up to 1.16% of total protein. To facilitate future purification of rVtg, we explored the possibility of secreting rVtg using the native Vtg secretion signal and the alpha factor secretion signal of Saccharomyces cerevisiae. However, neither signal promoted the secretion of rVtg. The clones maximally expressed rVtg at 23 degrees C, reaching a peak at 22 h in shake flasks and 16 h in a fermentor. The clones exhibited a significant increase in essential amino acids and long-chain polyunsaturated fatty acids, which are important for its application as a high quality nutrient feed. PMID- 14748090 TI - Protein disulfide isomerase, but not binding protein, overexpression enhances secretion of a non-disulfide-bonded protein in yeast. AB - In eukaryotes, secretory proteins are folded and assembled in the endoplasmic reticulum (ER). Many heterologous proteins are retained in the ER due to suboptimal folding conditions. We previously reported that heterologous secretion of Pyrococcus furiosus beta-glucosidase in Saccharomyces cerevisiae resulted in the accumulation of a large fraction of inactive beta-glucosidase in the ER. In this work, we determine the effect of introducing additional genes of ER-resident yeast proteins, Kar2p (binding protein [BiP]) and protein disulfide isomerase (PDI), on relieving this bottleneck. Single-copy expression of BiP and PDI worked synergistically to improve secretion by reverse similar 60%. In an effort to optimize BiP and PDI interactions, we created a library of beta-glucosidase expression strains that incorporated four combinations of constitutively or inducibly-expressed BiP and PDI genes integrated to random gene copynumbers in the yeast chromosome. Approximately 15% of the transformants screened had secretion level improvements higher than that seen with single BiP/PDI gene overexpression, and the highest secreting strain had threefold higher beta glucosidase levels than the control. Nineteen of the improved strains were re examined for beta-glucosidase secretion as well as BiP and PDI levels. Within the improved transformants BiP and PDI levels ranged sevenfold and tenfold over the control, respectively. Interestingly, increasing BiP levels decreased beta glucosidase secretion, whereas increasing PDI levels increased beta-glucosidase secretion. The action of PDI was unexpected because beta-glucosidase is not a disulfide-bonded protein. We suggest that PDI may be acting in a chaperone-like capacity or possibly creating mixed disulfides with the beta-glucosidase's lone cysteine residue during the folding and assembly process. PMID- 14748091 TI - Laboratory evolution of cytochrome p450 BM-3 monooxygenase for organic cosolvents. AB - Cytochrome p450 BM-3 (EC 1.14.14.1) catalyzes the hydroxylation and/or epoxidation of a broad range of substrates, including alkanes, alkenes, alcohols, fatty acids, amides, polyaromatic hydrocarbons, and heterocycles. For many of these notoriously water-insoluble compounds, p450 BM-3's K(m) values are in the millimolar range. Polar organic cosolvents are therefore added to increase substrate solubility and achieve high catalytic efficiency. Using p450 BM-3 as a catalyst for these important transformations requires that we improve its ability to tolerate the cosolvents. By directed evolution, we improved the activity of p450 BM-3 in the presence of dimethylsulfoxide (DMSO) and tetrahydrofuran (THF), achieving increases in specific activity up to 10-fold in 2% (v/v) THF and 6-fold in 25% (v/v) DMSO. The engineered p450 BM-3's are also significantly more resistant to acetone, acetonitrile, dimethylformamide, and ethanol as cosolvents in the reaction. PMID- 14748092 TI - A self-priming microfluidic diaphragm pump capable of recirculation fabricated by combining soft lithography and traditional machining. AB - Fluid transport is crucial in the development of microanalytical devices. While there are many micropump designs available, most are incapable of sustaining recirculation of fluid at microL/min to mL/min levels. We have designed and fabricated a positive displacement micropump by combining soft lithography with traditional bulk machining. The micropump is actuated through pneumatic pressure. The pump is self-priming and is suitable for recirculating fluid through a microfluidic device containing mammalian cell culture. By custom designing the volume of the pumping chamber, tight control of the output flow rate can be obtained by changing the actuation frequency. It can also be fabricated easily on plastic substrates without access to expensive microfabrication equipment. PMID- 14748093 TI - New classification and new diagnostic criteria of diabetes mellitus. PMID- 14748094 TI - A decade of growth in public and private pharmaceutical expenditures: the case of Belgium 1990-1999. AB - OBJECTIVE: To make a systematic, transparent, internationally comparable description of trends (1990-1999) in total, public and private (co-payment + out of-pocket) spending on pharmaceuticals in Belgium. SETTING: Belgium, a western European country, with a Bismarck-type universal coverage healthcare system. NATURE OF THE STUDY: Descriptive analysis of time-series. METHODS: Collaborative data gathering effort between academic and private research institutes and IMS health. RESULTS: Mean annual growth rate was 3.9% for total, 5.3% for public, and 2.0% for private drug expenditures (expressed in constant 1999 EUR). The ratio of public to private spending shifted from 53.4% to 60.3%. Of the private spending, one third was co-payment for reimbursed medication and two thirds was out-of pocket payment for non-reimbursed medication. CONCLUSION: Co-operation between several data gathering constituencies within one country was necessary to achieve completeness and detail in data collection on out-of-pocket payments for non reimbursed medicines, and hence in total drug expenditures. Discrepancies were found between the estimate of the public/private mix and OECD health data 2000 for public drug spending. PMID- 14748095 TI - Possible association between deep infection and hypertrophic osteoarthropathy. Report of three cases and review of the literature. AB - Hypertrophic Osteoarthropathy is classically associated with chronic pulmonary disease, cancer or inflammatory bowel disease. However, deep infection such as vascular graft infection is an etiology important to recognize because of the risk of life-threatening complication (aorto-enteric fistula). In this study, we reported two cases of aortic graft infection and one case of para-anal abscess associated with hypertrophic osteoarthropathy. The previously reported cases are reviewed. PMID- 14748096 TI - Actinomycosis as a rare cause of vertebral osteolysis. AB - Actinomycosis is an uncommon entity caused by an anaerobic bacterium, Actinomyces species, most often Actinomyces israelii. We present a patient who suffered from progressive lumbar pain and high fever. Actinomycosis infection was diagnosed after extensive radiological and pathological evaluation. Treatment with i.v. ampicillin was started, followed by oral antibiotics for one year. This paper gives an overview of the different clinical presentations of actinomycosis infection, with special attention to the skeletal involvement. We also discuss diagnosis and treatment. The clinical picture can mimic several other conditions, such as lymphomas. Delay in diagnosis and treatment can significantly worsen the condition of the patient. PMID- 14748097 TI - Renal extramedullary hematopoietic tumor diagnosed by fine needle aspiration: a case report. AB - We report a case of a 58 year-old-male with a history of sickle cell anemia, who presented with a left kidney mass. Guided fine needle aspiration of the mass revealed extramedullary hematopoiesis and enabled avoiding an unnecessary surgical procedure. INTRODUCTION: Extramedullary hematopoiesis (EMH) generally occurs in patients with deficient bone marrow hematopoiesis secondary to either peripheral red cell destruction or marrow replacement. EMH is most commonly seen in the liver and spleen as a diffuse lesion. Rarely EMH presents as a solitary mass, posing a diagnostic dilemma. In asymptomatic patients without obvious evidence of hemato-pathology, the differential diagnosis is even more complex. Despite several reports describing the radiographic findings in EMH, fewer promote the use of fine needle aspiration (FNA) in making this diagnosis. PMID- 14748098 TI - Cutaneous manifestations of internal diseases. AB - Collaboration between specialists is necessary for good practice of medicine. The dermatological examination, mainly based on visual characteristics, can greatly help other specialisms when confronted with a seemingly futile skin anomaly. We discuss the various cutaneous features of some metabolic and infectious diseases, the pitfalls in diagnosing lupus erythematosus without knowledge of skin disease and the relationship of dermatological diseases as cutaneous vasculitis, pyoderma gangrenosum, Sweet's syndrome, pemphigus and more to internal disease. PMID- 14748099 TI - Management of hypertension. AB - Presently, many drugs are available for the treatment of patients with high blood pressure. Although there are several theoretical arguments favouring one particular type of drug over the other, prospective clinical trials have failed to show major differences between drug classes in terms of overall prognosis. A possible exception in this regard may be the development of diabetes mellitus which seems to occur less frequently with ACE inhibitors, AT1 receptor blockers and calcium entry blockers than with diuretics. The choice of the antihypertensive drug regimen should take certain patient characteristics into account, notably comorbid conditions. However, despite theoretical considerations concerning initial therapy many patients will end up using two or more medications. PMID- 14748100 TI - Management of refractory systemic rheumatic diseases. AB - The management of refractory systemic rheumatic diseases remains a challenging issue. New immunosuppressive drugs have been developed which might improve long term outcome, with a reasonably toxicity profile. Moreover, better understanding of the mechanisms underlying some of the systemic rheumatic disease has raised hopes for more targeted immunointerventions. In Europe, current therapy of severe systemic rheumatic diseases (SRD) usually consists in a sequential treatment with a short course of an incisive remission-inducing immunosuppressive (IS) regimen [such as high dose glucocorticoids (GC) combined to cyclophosphamide (CYC)] followed by a long term but less toxic remission-maintaining IS regimen aimed at preventing relapses [such as low dose GC combined with azathioprine (AZA)]. This approach was recently supported by prospective studies performed in ANCA associated vasculitis (1) and lupus nephritis (2). However, significant number of patients with SRD fail to respond to this standard regimen and treatment of these refractory cases has become one of the many rheumatologist's nightmares. Interestingly, some recent advances in basic immunology, in our understanding of the pathogenic mechanisms operating in SRD and in biotechnology have opened new prospects, some of them already applied in clinical practice. In this paper, these features will be reviewed with systemic lupus erythematosus (SLE) as an example. PMID- 14748101 TI - Treatment of type 2 diabetes. AB - Type 2 diabetes is a heterogeneous disease resulting from a dynamic interaction between defects in insulin secretion and insulin action. As most subjects are overweighted or obese, the initial treatment is optimization of the meal plan and enhancement of physical activity in order to obtain sustained weight reduction. In case of failure of life-style changes, various oral antihyperglycaemic agents may be used. Some are targeting defective insulin secretion (sulphonylureas, glinides) while others are targeting insulin resistance (metformin, thiazolidinediones). Criteria of drug selection should include both patient's characteristics (body weight, age, degree of hyperglycaemia, comorbidities) and pharmacological properties of the compound (mode of action, safety profile, cost). Monotherapy is usually recommended first, but combined therapy using drugs with additive or synergistic effects may be required to obtain appropriate blood glucose control. As the natural history of the disease is characterized by a progressive exhaustion of beta cells, exogenous insulin may be required in the long term, usually in combination with oral agents. Finally, as patients with type 2 diabetes are insulin-resistant and often have a metabolic syndrome, a multifactorial intervention including aggressive treatment of arterial hypertension and dyslipidaemia is recommended in order to reduce the incidence of cardiovascular complications. PMID- 14748102 TI - How to deal with bone exposure and osteomyelitis: an overview. AB - The authors present an overview of the various techniques which can be used to achieve coverage of exposed bone, particularly in cases of bone exposure associated with an underlying fracture or osteomyelitis. Adequate debridement, possibly in several stages, is necessary in all cases to prepare the receptor site for the next step which is soft tissue coverage, achieved using various types of surgical procedures. Adequate reconstruction can be achieved with pedicled flaps in some cases but in cases with exposure of bone, free flaps usually represent a better option in cases where the condition of the patient is not a limiting factor. Thin fascio-cutaneous free flaps may be used in some cases with small and simple soft tissue loss, so as to minimise donor site mobidity. Free muscle flaps, such as from the latissimus dorsi, are preferable in cases with bone loss in order to fill any dead space; in cases with major bone loss, a free vascularised bone graft can be used, or composite grafts including bone, muscle and/or skin (fibula or crista iliaca flaps). Some reconstructions require a functional approach, such as over an exposed joint, or for the weight-bearing area of the foot or the soft tissues over the Achilles tendon. Survival of a free flap requires perfect, permeable microsutures; thrombosis of the anastomosis is a major complication which jeopardizes flap survival; close surveillance of the flap is required during the first few days, with hourly Doppler monitoring of the pedicle on the first day. The success rate can be as high as 90 to 100% in simple cases; failures may be related to surgical technique, inadequate choice of the flap, or specific features of the patient. In cases with an underlying bone infection, recurrence of infection occurs in 5 to 20% of cases; this requires additional treatment, possibly with repeat debridement, prolonged antibiotic therapy and sometimes a second free flap. PMID- 14748103 TI - The reversed Delta shoulder prosthesis in reconstruction of the proximal humerus after tumour resection. AB - The authors present two series of six and seven patients respectively, with a tumour of the proximal humerus, who were treated at two different institutions with a Delta type inverted shoulder prosthesis (DePuy International Ltd) after a Malawer type Ia or Ib resection. The rationale of using an inverted shoulder prosthesis is the aim to improve the functional outcome in rotator cuff deficient shoulders. This type of prosthesis medializes and lowers the centre of rotation, lengthens the lever arm of the deltoid muscle and improves its function. At one institution the resected part of the humerus was re-implanted after extracorporeal irradiation. It was fixed intramedullarly by cementation of the humeral prosthetic component to facilitate restoration of humeral height. This graft allowed reinsertion of muscles (deltoid, pectoralis, biceps) thus improving power generation postoperatively. The largest glenosphere, size 42, was routinely used to reconstruct the glenoid; this theoretically improves the functional outcome (increased external rotation) and stability. At the other institution no graft augmentation was used except in one patient. The height of the humeral prosthetic component was assessed after resection of the tumour by measurement of the resected part. The prosthetic stem was fitted in the remaining part of the humeral diaphysis, in three cases by cementation and in three cases by press-fit (hydroxyapatite coating). Muscle balance was appreciated intra-operatively. Stability of the prosthesis was directly related to the level of resection. Both techniques resulted in a minimum active abduction of 60 degrees, reaching 90 degrees or more in most patients. When compared to other results in the literature, this is a major functional improvement. The mean adjusted postoperative Constant score was 72.5% (range: 30-90%), and the mean MSTS score was 75.8% (range 36.7-96.7%). PMID- 14748104 TI - Souter-Strathclyde total elbow arthroplasty: medium-term results. AB - The Souter-Strathclyde unconstrained elbow prosthesis was prospectively studied in 36 patients (45 prostheses) with rheumatoid arthritis (Larsen grade 4 and grade 5). The mean age of the patients at the time of operation was 63 years (range: 39 to 75 years). Eight patients (9 prostheses) died within five years of implantation, from causes unrelated to the elbow arthroplasty. One patient was lost to follow-up, leaving 27 patients (35 prostheses) for review. The mean length of follow-up was 98 months (range: 60 to 174 months). At 8.2 years follow up, the prosthesis showed a probability of survival of 76% (SD 9%) with revision of the humeral component as an end point; the percentage dropped to 67% (SD 9%) when radiographic loosening was taken as an end-point. Survival of the ulnar component was 98%. Loosening of the humeral component seems to be related to both the short humeral stem and a persistent extension deficit. PMID- 14748105 TI - Implant removal on an outpatient basis. A patient satisfaction survey. AB - Unstable fractures of the distal radius can be stabilised with Kirschner wires. The K wires need to be removed after five to six weeks. The authors surveyed 100 consecutive patients with distal radial fractures treated with percutaneous Kirschner wires that were left buried under the skin. Group A had 50 patients who had such wires removed under local anaesthetic in the orthopaedic outpatient department. Group B comprised of patients who had the wires removed in the operating theatres under a short general anaesthetic. A questionnaire was designed to assess the satisfaction of these 100 patients. The survey showed that the patient satisfaction for this minor procedure carried out in the outpatient clinic was poor. Patient information and senior surgical input could increase the satisfaction. PMID- 14748106 TI - Outcome of distal radius fractures in relation to bone mineral density. AB - The purpose of this study was to determine whether osteoporosis influenced the clinical results of distal radius fractures in women above the age of 40 years. Forty patients underwent bone mineral densitometry of the distal forearm, hip and lumbar spine. Radiographs of both wrists were taken at the time of fracture healing and the difference in ulnar variance, palmar tilt and radial inclination with the contralateral wrist was measured. Wrist mobility, grip strength and pain relief were determined in 35 patients with a follow-up of more than one year. We found that the clinical results correlated better with bone mineral density than with the radiological parameters. Osteoporosis may be one of the factors affecting the outcome of distal radius fractures. PMID- 14748107 TI - Chronic lunotriquetral dissociation: dorsal capsular reinforcement with a retinaculum flap. AB - The authors have used dorsal capsular reinforcement with a retinacular flap to treat 13 patients with chronic lunotriquetral instability. The patients, 8 female and 5 male with a mean age of 30 years, were reviewed by an independent observer after a mean follow-up of 3.2 years. Overall patient satisfaction was poor in five, moderate in five and excellent in three. Three patients underwent a secondary procedure: lunotriquetral arthrodesis in two and tenolysis of the extensors in one. Overall, the authors did not find a significantly better outcome compared to lunotriquetral arthodesis, but the complication rate and the rate of reoperations were lower. PMID- 14748108 TI - Intra-articular bupivacaine hip injection in differentiation of coxarthrosis from referred thigh pain: a 10 year study. AB - The authors conducted a retrospective study to validate the specificity of intra articular injection of local anaesthetic to identify the source of pain in patients with coxarthrosis but ill-defined clinical and radiological features. Forty-seven patients received intra-articular injection of the hip joint as a diagnostic procedure. Twenty-four patients showed a positive response with relief of pain. All of them underwent total hip replacement and remained pain free following surgery. In twenty-three patients intra-articular injection did not relieve the pain; three of these underwent successful total hip replacement two years later. Other negative responders were further evaluated and appropriately managed. This study confirms that intra-articular injection of local anaesthetic is a valuable tool in diagnostic dilemma. The calculated sensitivity of 88% and specificity of 100% is similar to other published series. PMID- 14748109 TI - Inferior dislocation of the proximal tibiofibular joint: a report on four cases. AB - The authors report four cases of inferior dislocation of the proximal tibiofibular joint. A literature search disclosed reports on superior, anterolateral and posterolateral dislocation, but none on inferior dislocation of the proximal tibiofibular joint. The latter was associated in these four cases with fracture of the tibia, as also noted in cases of superior dislocation, and with severe neurovascular lesions. PMID- 14748110 TI - Influence of burst TENS stimulation on the healing of Achilles tendon suture in man. AB - Retrograde or antidromic stimulation of the nociceptive C fibres is known to lead to the release of sensory neuropeptides Substance P (SP) and Calcitonin Gene Related Peptide (CGRP) by the peripheral endings of these ultra-thin nerve fibres. These neuropeptides have, among others, a vasodilatory effect, which explains why they play a role in the healing of soft tissues. Burst TENS (Transcutaneous Electric Nerve Stimulation) is known to be most effective in influencing C fibre-evoked activity. This is why burst TENS was used in a randomised study as a stimulus for the healing of the sutured Achilles tendon in 10 patients, versus 10 others who received no stimulus. There was one drop-out in each group, so that 2 x 9 patients remained available for the study. A needle biopsy, performed after six weeks, showed no significant influence of burst TENS on the histological healing stage, as compared with a rat study. However, a semi quantitative evaluation of the number of fibroblasts showed a significant advantage for the stimulated group: p = 0.007. This means that burst TENS might influence healing of Achilles tendon sutures in man. But above all, it means that a histochemical study of the influence of burst TENS on the release of substance P and CGRP, after suture of the Achilles tendon in man, would be worthwhile. PMID- 14748111 TI - Stage I compression-extension injury of the cervical spine. A rotationally unstable injury. AB - Stage I compression-extension injury of the cervical spine, as described by Allen et al, is not always a stable injury. The combined unilateral failure of the posterior structures under compression together with failure of the anterior structures under tension will lead to rotational instability around the intact lateral mass. We report on 10 consecutive patients who presented with this type of injury. The surgical protocol consisted of early reduction followed by anterior cervical fusion using a tricortical iliac graft and a locking plate. Mean follow-up was 38.5 months. Intra-operative assessment revealed disc injury in all patients. Anatomical realignment and solid fusion were achieved in all cases. All 10 patients showed improvement of their neurological deficit. One patient remained with some residual weakness in his triceps, and another required removal of a prominent screw. PMID- 14748112 TI - Functional reconstruction of the upper extremity in tetraplegia. Application of Moberg's and Allieu's procedures. AB - The authors conducted a retrospective study to evaluate the benefit of 43 reconstructive procedures and tendon transfers, performed in 37 sessions to restore hand and arm function in 25 tetraplegic patients. These operative procedures were either single or multiple, depending on the patients' needs for restoration. The initial clinical situation was assessed based on a simplified version of the international classification of Giens, modified in Edinburgh. The results were evaluated through clinical assessment and simple functional testing. Gestural ability was improved in more than 80% of the patients and functional gain was important in more than half. The authors conclude that selected tetraplegic patients can benefit from these procedures. The diversity of clinical situations and the variety of surgical procedures result into small scattered series unfit for statistical analysis, and there is a need for unequivocal, internationally recognised assessment methods. PMID- 14748113 TI - Evaluation of oxidative stress after fractures. A preliminary study. AB - In a preliminary attempt to see if oxidative stress occurs after major fractures, we evaluated two groups of patients sustaining a single fracture (Group A) and multiple fractures (Group B), and compared them with healthy controls (Group C). Indirect evaluation using plasma was done, as serial samples directly from bone could not be taken in humans. We measured plasma levels of malonyldialdehyde (MDA), which depicts the lipid peroxide content, and the unstable nitric Acid (indirectly through measuring reactive nitrogen intermediates and citrulline), serially over a four-week period. The endogenous ferric reducing anti-oxidant power assay (FRAP) was also done. All these have been proven to be representative of oxidative stress in other situations. We noted significant changes in these values, peaking by the 2nd and 3rd weeks post fracture, and coming down by the 4th week. This implies that oxidative stress does occur after a fracture; the initial few days are eventless, probably because the fracture causes a local region of ischaemia. Oxidant levels rise by the 2nd and 3rd week, perhaps due to callus formation and angiogenesis, which results in reperfusion at the fracture site. Oxidative stress may also be proportional to the number of bones fractured, as Group B, with multiple fractures had more elevated values. The antioxidant levels also behave similarly to combat the detrimental effect. The pro and antioxidants values then gradually decline by the 4th week, probably because by then the bone starts to organize. A better understanding of these mechanisms may help in defining the role of oxidative stresses after fracture and perhaps better define the role of antioxidants in helping fracture healing. PMID- 14748114 TI - Bilateral congenital coracoclavicular joint. Case report and review of the literature. AB - The diagnosis of coracoclavicular articulation was made in an elderly woman, who complained of bilateral anterior shoulder pain with impingement. This congenital pseudo-articulation is rarely symptomatic and is usually an incidental radiological finding. Surgical excision of the prominent bony projection, arising from the inferior surface of the clavicle at the level of the coracoid process, relieved the impingement and gave satisfactory pain relief. PMID- 14748115 TI - Acute subclavian artery pseudoaneurysm after closed fracture of the clavicle. AB - A sixty-year-old woman was admitted with an acute subclavian artery pseudoaneurysm after a closed fracture of the middle third of the right clavicle. This manifested itself a few days after the injury as a slowly growing, pulsatile and purple tumour-like mass in the right supraclavicular fossa. The distal pulses were normal, but this observation does not exclude a vascular trauma. This is an uncommon but potentially dangerous complication as it jeopardizes both the extremity and the life of the patient. PMID- 14748116 TI - Multidirectional voluntary glenohumeral dislocation in a 7-year-old patient: a case report. AB - A case of multidirectional voluntary shoulder dislocation developing at the age of seven is reported. The authors present the process that leads to the diagnosis of voluntary shoulder dislocation. The treatment of young patients with this condition is difficult. If no underlying pathology exists, most authors agree it should consist of conservative treatment and skillful neglect. The role of surgical treatment in a maturing skeleton should be reserved only if conservative treatment fails. PMID- 14748117 TI - Use of a Dall-Miles plate and cables for the fixation of a periprosthetic humeral fracture. AB - Increase in the use of shoulder arthroplasty has resulted in the emergence of periprosthetic fractures of the humerus. The management of such fractures is technically demanding. We describe a case in which a displaced periprosthetic fracture at the tip of a stable humeral implant was successfully managed with the Dall-Miles plate and cables system. PMID- 14748118 TI - Presentation of compartment syndrome without an obvious cause can delay treatment. A case report. AB - Compartment syndrome is a serious condition which leads to chronic morbidity unless an urgent decompression of the affected area is performed. An increased intra compartmental pressure commonly occurs after a physical insult though rarer causes have been identified. We report an atypical presentation of compartment syndrome and subsequent delayed intervention where there was no identifiable aetiological factor. Frontline medical staff must rule out compartment syndrome early so that complications secondary to compartment syndrome can be avoided. PMID- 14748119 TI - Compression of the medial branch of the deep peroneal nerve, relieved by excision of an os intermetatarseum. A case report. AB - The authors report a case of direct compression of the medial branch of the deep peroneal nerve by an os intermetatarseum in a 52-year-old female patient who was referred to their Institution because of pain over the dorsum of her left foot associated with paraesthesias in the first web space. Examination disclosed a positive Tinel sign over the dorsal aspect of the first metatarsal bone. Plain radiographs revealed a small, irregular accessory ossicle on the dorsum of the left foot, between the medial cuneiform and first and second metatarsals. At operation, the os intermetatarseum was found to impinge on the medical branch of the deep peroneal nerve. Excision of the os intermetatarseum and nerve decompression was performed. After four years, the patient has normal function and is completely relieved of her symptoms. PMID- 14748120 TI - The way forward? PMID- 14748121 TI - Working in partnership: the breastfeeding agenda. PMID- 14748122 TI - Trials and tribulations. PMID- 14748123 TI - The heart of the matter. PMID- 14748124 TI - Changing attitudes and perceptions to hyperemesis gravidarum. AB - A small, but significant proportion of pregnant women (1% to 2% of all pregnancies), suffer from a severe form of nausea and vomiting, which is categorised as hyperemesis gravidarum (HG). The arbitrary definition, aetiology and management of HG, which leads to conflicting attitudes and beliefs among healthcare professionals, inevitably impacts on the woman's overall experience of the condition. It causes great distress and if medical and ward staff misunderstand the condition, it can increase rather than reduce the suffering of women. A structured review was designed, both to determine the attitude and beliefs held by healthcare professionals and to advance understanding of women's experiences in relation to HG. A comprehensive search of literature was carried out. Due to the nature of the study, all the relevant identified and included articles were qualitative, therefore narrative synthesis of the tabulated results was carried out. The findings are discussed under the following themes: the experiences of the women, the various attitudes of health professionals. How the relationship between patients and health professionals is in regard to the mother's satisfaction with care and treatment. In the absence of evidence for an effective treatment, it is crucial to try and understand this complex condition, and to expand the circle of support from family to healthcare professionals involved in looking after sufferers. PMID- 14748125 TI - PPF targets and breastfeeding initiation rates. PMID- 14748127 TI - Breastfeeding--where has the common sense gone? PMID- 14748126 TI - Policy, practice and research: does it make a difference? PMID- 14748128 TI - Transmitting data speeds angioplasty for MI patients. PMID- 14748129 TI - 'Immediate bedding' boosts patient satisfaction at California emergency department. AB - For sick or injured patients, the average wait of nearly an hour to see a physician at Good Samaritan Hospital's ED must have seemed like an eternity. But since instituting a practice known as "immediate bedding", unhappy patients have almost disappeared and its patient satisfaction ratings have skyrocketed. PMID- 14748130 TI - Home care agency implements pain management procedures. AB - Pain management was an often overlooked aspect of health care until 2000, when the JCAHO included standards for assessing and managing pain in its accreditation criteria. Hospitals, home care agencies, nursing homes, and other organizations suddenly found themselves with a large need to educate employees and patients about effective pain management techniques. PMID- 14748131 TI - Whole practice productivity applies 'big picture' solution to performance improvement. AB - When health care organizations look at improving productivity, they typically focus on a few specific measures--work RVUs, office visits per day, or gross charges per physician, for instance--in which they've identified variations among physicians or clinics. Instead of relying strictly on the collection of a handful of isolated data points, two consultants affiliated with the Medical Group Management Association are trying to convince provider groups to become more efficient by looking at the bigger picture. PMID- 14748132 TI - Managers can take these four steps to reduce wasteful work. PMID- 14748133 TI - [The structure and function of nicotinic acetylcholine receptor]. PMID- 14748134 TI - [Structural biology of the interleukins]. PMID- 14748135 TI - [Discovery, analysis and chemical mechanism of protein splicing]. PMID- 14748136 TI - [Photoperiodic control of flowering: molecular mechanisms to diverse long-day plants and short-day plants]. PMID- 14748137 TI - [A novel mechanism that regulates the start of cortical cell migration out of the ventricular zone]. PMID- 14748138 TI - [Hans Krebs, discoverer of metabolic cycles]. PMID- 14748139 TI - [Dr. Shinya Inoue and Shinya-scope]. PMID- 14748140 TI - [Trends in bioinformatics]. PMID- 14748142 TI - [Why patent now?]. PMID- 14748141 TI - [Bioinformatics in Asia]. PMID- 14748143 TI - [Homeodomain transcription factor T/EBP/NKX2.1 in development and differentiation of the thyroid and lung]. PMID- 14748145 TI - [Folding of protein (lysozyme)]. PMID- 14748144 TI - [Regulation of B cell receptor signaling]. PMID- 14748146 TI - [Frequent occurrence and biological significance of degree of polymerization in sialic acid]. PMID- 14748147 TI - [Neurosteroids are 4th generation neuromessengers which are synthesized and enhance/suppress learning and memory in the brain]. PMID- 14748148 TI - [Signaling mechanism in the regulation of osteoclast differentiation by the immune system]. PMID- 14748150 TI - [Carbohydrate microarray: a sweet spot for deciphering the information embedded in oligosaccharide structures]. PMID- 14748149 TI - [Crystal structure of the core domain of troponin and the mechanism of muscle regulation]. PMID- 14748151 TI - [Recombinant antibodies: basics and application to medicine]. PMID- 14748152 TI - Guide to care for patients. HPV & cervical cancer. PMID- 14748154 TI - Cervical cancer: what every woman needs to know. PMID- 14748153 TI - Urinary incontinence. Solving a secret problem. PMID- 14748155 TI - Understanding and treating PMS/PMDD. PMID- 14748156 TI - Diabetes update. PMID- 14748157 TI - Nursing leadership in health care: WHO nursing midwifery services strategic directions. PMID- 14748158 TI - The future nursing voice. AB - Based on some articles in the journal Nursing Ethics, the author outlines some of the areas of major importance for nursing in the future. These areas--the care of elderly people, long-term home-based care, genetics, international research and conflict and war--demand a new voice of nursing, which is a political voice. The rationale for a political voice is the ICN Code of ethics for nurses and the fourfold responsibilities laid on nurses: to promote health, to prevent illness, to restore health, and to alleviate suffering. Some indications are given on how nurses can engage in political work. PMID- 14748159 TI - [Epistemological constructs in sciences -- a contribution to nursing]. AB - This study presents some critical reflections "about epistemological constructs in science", on the basis of concepts and preoccupations that are inherent to the scientific and technological advances that influence the construction of knowledge. In accordance with the methodological approach, nursing is seen as "a science under construction". The discussion and arguments focus on distinct aspects/traces of science, which define the nature and objectives of research and which broaden the view of objective knowledge and reality for the field of nursing. In coherence with philosophical and academic reference bases, the author uses the focus of discourse to emphasize/highlight the meaning of constructs as being consistent with the elaboration of significant theories or issues in a field of study and explains her intellectual position in relation to some nursing theses (Ph.D.), in an attempt to explain epistemological aspects of the construction of nursing science. PMID- 14748160 TI - [Intuitive ability in nursing care]. AB - This study aimed to understand the intuitive ability of nursing professionals in caring. The study method was qualitative, adopting a descriptive exploratory approach. Sampling involved 87 female nursing professionals, being 31 nurses, 29 nursing technicians and 18 nursing auxiliaries. The result showed different levels of intuitive abilities, related to novice, standard, and veteran. The relation between the three levels of intuitive abilities and the corresponding figures in years, observed in a North American study, was not identified. The factors described as affecting the intuitive experience were both environmental and intra-interpersonal (personological). The experiences of the professionals portrayed the importance of intuition in nursing care, mainly in doubtful and conflicting situations and, in this sense, its vital function was decision-making towards better quality of nursing care. PMID- 14748161 TI - [Hospitalization in neonatal intensive care units in Brazil -- 1998-2001]. AB - This study aimed at obtaining information on the development of data records in Neonatal Intensive Care Units in Brazil; comparing the number of beds in Neonatal Intensive Care Units with that in Intensive Care Units in general and assessing their distribution in different Brazilian states. There has been an increase in the number of beds and authorizations for hospitalizations in intensive care units. The percentages indicating the total number of beds for intensive care and neonatal intensive care vary from one state to another with 17% in Sao Paulo, 25% in Rio Grande do Sul and 29.5% in Parana. Sao Paulo state presents the highest number of beds, followed by Rio Grande do Sul. PMID- 14748162 TI - [Perception of parents in experiencing the kangaroo mother method]. AB - This study aimed at analyzing the perception of pre-term babies' parents in experiencing the Kangaroo Mother Method. It is a descriptive study with a qualitative approach. Data were collected from semi-structured interviews with 10 parents, held in the first 60 days after the discharge of the pre-term baby from the Kangaroo method, in a philanthropic hospital in the interior of Sao Paulo State. The parents' perception concerning the experience was grouped in four thematic units: making maternal stay in the Kangaroo Method flexible; favoring the mother-child and family relationship; completing the pre-term newborn's growth and development and developing skills in caring for the child. The obtained data provided information for the organization of care in the Kangaroo Mother Method, from the institutional perspective of the health care team as well as the relationship with clients, which enabled understanding about the difficulties and meanings attributed to the experience and optimization of nursing care. PMID- 14748163 TI - [Teacher and student sharing the teaching-learning experience: a pediatric nursing course at the Federal University of Mato Gross do Sul]. AB - The authors present the experience of teaching pediatric nursing at the Universidade Federal de Mato Grosso do Sul (UFMS), Brazil. They carried out a qualitative analysis of the learners' evaluation for the year 2000, from which two categories emerged: "being a different experience" and "changing the way the learners see the child and the family". They conclude that choosing the person centered teaching approach has been of fundamental importance for the learner to achieve personal and professional autonomy. PMID- 14748164 TI - [Characterization of obesity in patients with myocardial infarction]. AB - This study aimed to characterize obesity in infarcted myocardial patients. Data were collected from 43 patients at a University Hospital. It was observed that the Body Mass Index (BMI) was above the desired level for 69.7% (26.9 +/- 0.5) of the subjects; the body fat percentage had altered in 79% (20.0 +/- 1.0) of cases; the waist/hip (W/H) ratio was above normal in 72.5% (0.97 +/- 0.03) of cases and the waist circumference (WC) had altered in 64% (97.3 +/- 1.8) of the patients. Since the correlation for BMI, WC and fat percentage was significant, the use of BMI and WC was suggested for evaluating obesity in outpatients. PMID- 14748165 TI - [Degree of patient dependence in relation to the nursing staff: analysis of clinical records]. AB - In this study, we intend to identify in the evolution of nursing the intensity of patient dependency as it is written down by the nurse and confront it with a ranking instrument. 74 patients, 11 nurses and 04 lecturers were part of the study held from September 22 until October 27, 2000. We noticed slight modifications in the degree of patient dependency from hospital admission to discharge. Furthermore, the majority of the patients that needed help were the intermediate ones (41.9%). However, our reality still requires nursing training and the creation of an appropriate classification instrument, in accordance with the features of our internment units. PMID- 14748166 TI - [The meaning of specialization for the nurse in the field of stomas]. AB - This study aimed at knowing and understanding the meanings and perceptions of the enterostomal therapist (ET) about this specialty, using the methodological framework of Social Representation Theory. Three major Units were identified: the ET as a professional, the ET and the client, the ET as a person from an analysis of 12 ET's discourse. In relation to Enterostomal therapy, mainly positive representations were identified among nurses, such as: valorization, recognition, remuneration, promotion, satisfaction, ambition, professional prominence, improvement of care quality and client safety. On the other hand, demands and lack of recognition emerged as negative representations. These representations indicate specialty as an important dimension in professional evolution, a fact that has positively influenced the career of the nurse and that of the Enterostomal Therapist in particular. The ET's construct their existence, write their own history, driven towards autonomous and credible action, which allows them to conquer their space and give a new meaning to care--the basis of Nursing art and science. PMID- 14748167 TI - [Alternative/complementary therapies in public and private education: analysis of knowledge among nursing students]. AB - This study aimed to analyze undergraduate Nursing students' knowledge in two education institutions, one public and one private, about alternative/complementary therapies, since both institutions offer a specific subject in this area under the optional and obligatory form, respectively. Data suggest that, apart from academic education, knowledge about this theme originates from common sense. The students recommend the use of alternative/complementary therapies although they do not use them much. Flower therapy, acupuncture, homeopathy, cromotherapy, fitotherapy, music therapy and massage are more utilized. The most neglected aspect in the teaching-learning process refers to the legal aspects of specialization in this area for the nurse. PMID- 14748168 TI - [Leisure, life besides work for a soccer team of hospital workers]. AB - With a view to promoting workers' mental health, minimizing professional stress and fatigue, we investigate the meaning of work, leisure and their implications for 24 male workers at a Sao Paulo hospital school, who are part of a soccer team and attend the recreation association of this unit. We chose to realize Research Action from a humanist, qualitative approach, analyzing discourse according to categories, to be of help in elaborating the educational project. Data were collected through interviews and participant observation, using photography as a support instrument. Most participants are married, have children, receive an average salary of R$650.00 and work near the unit, which favors contact for games. In their opinion, work guarantees personal and family survival, while leisure means fun, relaxing, integration and valorizing family and friends. They emphasize the importance of challenge in soccer as a means of promoting health, rescuing self-esteem, happiness, freedom, creativity, spontaneity, resulting in better preparation for personal and professional life. PMID- 14748169 TI - [Creation and management of organizational knowledge]. AB - With a view to creating and establishing a sustainable position of competitive advantage, the best organizations are increasingly investing in the application of concepts such as learning, knowledge and competency. The organization's creation or acquisition of knowledge about its actions represents an intangible resource that is capable of conferring a competitive advantage upon them. This knowledge derives from interactions developed in learning processes that occur in the organizational environment. The more specific characteristics this knowledge demonstrates in relation to the organization, the more it will become the foundation of its core competencies and, consequently, an important strategic asset. This article emphasizes nurses' role in the process of knowledge management, placing them in the intersection between horizontal and vertical information levels as well as in the creation of a sustainable competitive advantage. Authors believe that this contribution may represent an opportunity for a reflection about its implications for the scenarious of health and nursing practices. PMID- 14748170 TI - [Long-acting antipsychotics in the maintenance treatment of schizophrenia. Part II. Management of medications, integration of the multiprofessional team, and perspectives with the formulation of new a new generation of long-acting antipsychotics]. AB - Among various topics, this second part addresses indication and beginning of treatment, dose inter-individual variability and interval between injections, appointment frequency and special strategies during treatment relapse. Considering that poor adherence to antipsychotic treatment is a major factor in schizophrenic relapse, that the new generation of antipsychotic drugs, despite lower incidence of extrapyramidal side effects and better overall tolerability, did not change this condition in relation to conventional drugs and in view of the superiority of depot antipsychotics in comparison with conventional ones administered orally, the long-acting formulation of new generation antipsychotics can certainly improve the adherence and regularity of the medication regimen and decrease relapse rates in patients with schizophrenia. Furthermore, family participation in treatment is of great importance, as well as the attitude and integration of the medical team in realizing different tasks. PMID- 14748171 TI - [Care protocols for diseases caused by bioterrorism]. AB - Epidemics due to bioterrorist actions and prevention and control measures of these infections have been worrying not only the policy makers but also health professionals in general. Health professionals have to recognize these diseases earlier. Information and updated studies about the issue are needed. This study is a literature review in the MEDLINE and LILACS databases, books and online information (from 1993 to 2001) about epidemic aspects of the main diseases due to bioterrorist actions and care recommendations for infection control and occupational risk minimization. Healthcare professionals, who assist patients directly, should be prepared to assist patients who become ill in isolated or epidemic situations. PMID- 14748172 TI - [Interdisciplinarity and health: bibliographic study]. AB - This is a bibliographic research which sought to achieve a better understanding of the interdisciplinarity theme. The complexity that characterizes the world and especially the health area nowadays has required the development of interdisciplinary teaching programs, with a view to obtaining a new kind of thinking and the formation of health professionals committed to social reconstruction. We examined literature for the meaning of interdisciplinarity, its history and relations with Collective Health and the education of health professionals. PMID- 14748173 TI - [Research on quality of life: review of the scientific production of public universities from the state of Sao Paulo]. AB - This descriptive and exploratory study aims to analyze the scientific production regarding quality of life by the public universities in Sao Paulo State on the basis of data collected from the elected universities' virtual libraries. The preliminary analysis resulted in 84 studies: 71.4% were Master's Theses, 23.8% doctoral dissertations and 4.7% associate professor's theses from 1993 to 2001. 1 studies were excluded from this total. The 53 (100%) remaining studies had been developed with male and female adults (71.7%) who presented some type of pathology (69.8%) and used existing instruments to assess quality of life (77.3%). The most frequently used instrument was the Medical Outcomes Studies 36 item Short-Form (MOS SF-36), found in 18 (34%) of these studies. PMID- 14748174 TI - [Encouraging mother-child attachment in premature situations: nursing interventions at the Ribeirao Preto clinical hospital]. AB - This study aims at describing nursing actions performed in the high-risk neonatal units at a university hospital of the University of Sao Paulo at Ribeirao Preto so as to favor mother-child attachment in prematurity situations. The nurse accompanies parents in their first visit, giving them support as well as information concerning the equipment surrounding the newborn and encouraging skin to-skin contact, touching and talking. Parents' access to and staying with high risk newborns is permanently allowed. A visiting program by grandparents and siblings of pre-term newborns was implemented, even when under intensive care, which encourages family contact. Parents participate in a support group with other parents who experienced the situation of having their pre-term children in serious conditions and hospitalized. We consider that our experience has favored the establishment of mother-child and family attachment, observing greater interaction between the family and the newborn, particular involving the mother. Greater interest in learning about care as well as satisfaction concerning the assistance received have also been expressed by families. PMID- 14748176 TI - [Battered wives]. PMID- 14748175 TI - [Nursing practice in the health care of children at a basic health unit]. AB - This study aimed at identifying nursing actions in the follow-up of growth and development of children under five years old who were assisted at two Basic Health Units in Franca, Brazil. The empirical study was based on observation of nursing activities in the vaccination, pre-consultation, post-consultation and newborn screening test collection sectors, using video recording. We concluded that basic follow-up actions concerning growth and development, breastfeeding stimulation, food orientation, vaccination and prevalent disease prevention are permeating nursing care, although in a fragmented way in communication and ties established with clients. PMID- 14748177 TI - [Detection of early stage bronchopulmonary cancer: contribution of bronchoscopy]. AB - Efficient methods for lung cancer screening is today an important challenge in research. Into this article, we review the available techniques for the detection of hilar located lung cancers, mainly squamous cell carcinoma. Fluorescence bronchoscopy is the latest major advance for lung cancer diagnosis. PMID- 14748178 TI - [Lateral peritalar luxation: prognostic evaluation and therapeutic approach. Review of the literature based on a clinical case]. AB - The incidence of the peritalar dislocations has been estimated to be approximately 1% of all dislocations. If they are missed, the consequences are serious and lead to an important surgery of the hindfoot. Lateral dislocation is most rare than medial dislocation. The diagnosis of this lesion often requires X rays of the foot. The CT-scan allows to confirm the diagnosis and to appreciate the associated intra-articular fracture. The reduction must be realized in urgency under anaesthesia. If the orthopaedic reduction is impossible, the surgery allows to obtain an anatomical reduction with the removal of obstacles and the fixation of the associated intra-articular fractures. The prognosis of this lesion is better if there is an appropriate and rapid treatment. PMID- 14748179 TI - [Psychostimulants: a potential therapeutic alternative in mood and obsessive compulsive disorders (first part)]. AB - Among illicit substances in all the world, psychostimulants fill the second place. Their bad use has greatly increased at the end of the eighties in parallel to the increases of varied party and to the research of putting up a good physical or intellectual performance. But their employment is not recent as it appears by their use at large scale during the Second world War and after it in Japanese society. A MEDLINE search from 1963 to 2002 was done to identify the literature published on psychostimulants and their sociocultural use, their basic chemistry, their pharmacology and more specifically their psychiatric uses and indications in thymic and obsessive compulsive disorders. The references of articles found were evaluated for other relevant articles. PMID- 14748180 TI - [Endoscopic percutaneous gastrostomy]. AB - Enteral nutrition is indicated in patients with a functional bowel who are unable to orally ingest sufficient amounts of food. Nasogastric tube feeding is commonly used, but for patients requiring long-term tube feeding, percutaneous gastrostomy must be considered. Indeed, this technique is safe and well-tolerated. The decision of placing a percutaneous endoscopic gastrostomy should take into account medical, nutritional, and ethical considerations. PMID- 14748181 TI - [Authorized publicity? Amendment of the Code of Ethics by the National Council of the Order of Physicians on the publicity and three related notices]. AB - In the issue of december 2002 of the bulletin of the National Council of Medicine are detailed some amendments about the chapter "PUB and publicity" of the Code of Ethics and two closely related notices. Another notice about neon signs take place in the bulletin of june 2003. Globally, the National Council does not authorize PUB but adopt a more moderate position about publicity in a series of particular situations. This article discuss the new situation and what it is necessary to think about. PMID- 14748182 TI - [Topical immunosuppressors: new therapeutic approach in atopic dermatitis]. AB - The efficiency of topical immunosuppressors in the treatment of atopic dermatitis is undebatable. These agents induce immunosuppression by inhibiting T cell activation and cytokine production. An intermittent treatment, with two applications per day, and photoprotection is recommended in adults and children aged more than two years. The precise position of the topical immunosuppressors in the treatment strategy of atopic dermatitis remains to be defined. Main side effects are irritative or allergic cutaneous reactions and cutaneous infections, no systemic side effects have been recorded. The consequence of long-term topical immunosuppression and specially the possible development of skin cancers in UV exposed skin, is not well known and necessitates careful monitoring of patients. These treatments are contra-indicated during pregnancy, breast feeding, in erythrodermic states and with any alterations of the cutaneous barrier. PMID- 14748183 TI - [Comment on the article written by P.G. Silance published in Rev Med Brux 2003; 24: A253-6]. PMID- 14748184 TI - [In reference to the J.E.P. 2003: comments on "Electrocardiograms and clinical cases"]. PMID- 14748185 TI - [Artificial nutrition: from the hospital to the home]. PMID- 14748186 TI - [Home enteral nutrition]. PMID- 14748187 TI - [Home parenteral nutrition]. PMID- 14748188 TI - [Ethics and artificial nutrition]. PMID- 14748189 TI - [Minnesota 1954: the spring of modern heart surgery. Homage to Clarence Walton Lillehei (1918-1999)]. PMID- 14748190 TI - [Image of the month. Duodenal varices complicating portal hypertension]. PMID- 14748191 TI - [Image of the month. Traumatic diaphragmatic hernia]. PMID- 14748192 TI - [How I treat ... essential tremor]. AB - Essential tremor is the most prevalent movement disorder. This condition is often considered as benign but it can be the cause of considerable social, professional, and psychological handicap. It affects people of all ages but its prevalence increases with age (up to 5% of patients 65 or older). After discussing clinical aspects of this disorder, we review the current medical and surgical therapeutic options. Surgery may bring up spectacular improvement. PMID- 14748193 TI - [Clinical case of the month. Penetrating trauma of the thoracic aorta caused by a deer horn]. AB - We present a 33 year-old patient who sustained a penetrating wound of the thoracic aorta caused by a deer horn, without any other lesions. A brief survey of the literature summarizes the epidemiology, diagnosis, surgical care and prognosis of this lesion. PMID- 14748194 TI - [Role of kinesitherapy in the treatment of vertigo]. AB - The author describes how to use kinesitherapy in the treatment of vertigo, particularly benign positional vertigo and vertigo due to labyrinthine lesions. PMID- 14748195 TI - [Problematic resistance of pneumococcal antibiotic resistance]. AB - Streptococcus pneumoniae is the most common cause of community-acquired pneumonia and it is also a common cause of sinusitis, otitis media, bacteremia and meningitis. The increasing resistance to antimicrobial agents, now endemic in many countries, reflects an uncontrolled use of antibiotics. A good antibiotics policy and vaccination are at the moment the only way to control efficaciously the increasing antibiotic resistance of Streptococcus pneumoniae. PMID- 14748196 TI - [Effects of androgen substitution on lipid profiles in hypogonadal men]. AB - The decrease in serum bioavailable testosterone level may be responsible for the catabolic sequelae noticed in the ageing male (decrease in libido, decrease in muscle mass, osteoporosis and increase in adiposity). After brief review of androgens and lipids metabolism as well as their modifications with ageing, we propose to present the current knowledge of the effects of androgen substitution on lipid profile in hypogonadal men. PMID- 14748197 TI - [Synopsis of invasive dermatomycoses in immunocompromised host]. AB - The immune system plays a prominent role in the defense against a wide variety of pathogen fungi. Thus, the immunocompromised patient becomes a potential victim of mycoses originating from the skin. These mycoses may become invasive and a fatal issue is possible. In other circumstances, the inoculation site is pulmonary or digestive, and the cutaneous lesions represent secondary localizations. PMID- 14748198 TI - [Coverage of defects: principles]. AB - The coverage of defects is a broad field with which the plastic surgeon is confronted daily within traumatic, tumoral or other context. The various techniques used are skin graft and flaps, forming both heterogeneous groups. Indeed, there are various types of skin graft although a common denominator is the need for a good recipient site in order to allows an adequate "take". On the other hand, flaps carry their own vascularization. Thus, they are not dependent of the recipient site for their survival. Those are divided into three groups: local flaps, pedicled flaps and free flaps. The choice of the adequate technique with respect to the defect to be covered depends on the characteristics of the defect, its localization, the functional requirements of the area, the exposed structures, the medical status of the patient. The possible morbidity left on the donor site the aesthetic and functional goals are taken into consideration. For each case, there are often several good options as well as others less optimal solutions. The existing solutions are often so numerous that the plastic surgeon is frequently able to solve all the types of defects. PMID- 14748199 TI - [How I explore ... the controversy concerning the role of oral glucose tolerance test in clinical practice]. AB - The oral glucose tolerance test (OGTT) has been widely used for the diagnosis of diabetes mellitus, gestational diabetes or impaired glucose tolerance. However, since 1997, OGTT is not considered anymore as the first choice test for the diagnosis of diabetes mellitus or impaired glucose tolerance, being replaced by fasting glycaemia. Numerous studies, however, stressed the discrepancies between these two diagnostic approaches, so that OGTT still has defenders as a useful tool to diagnose diabetes. Besides, emphasis has been recently put on both postprandial hyperglycaemia, considered as a major cardiovascular risk factor, and impaired glucose tolerance as a predictor of progression toward overt type 2 diabetes, two parameters well addressed by the OGTT. Thus, even if the place to be reserved to the OGTT is controversial, in our opinion, this test still has a place in clinical biology. PMID- 14748200 TI - [Medication of the month. Tazarotene 0.05%-0.1% (Zorac)]. AB - Since April 28th, a new topical retinoid is available in Belgium: tazarotene (Zorac) gel (Pierre Fabre). It exists at two concentrations: 0.05% or 0.1%. It is the first receptor-selective topical retinoid. It is indicated in the treatment of plaque-type psoriasis. In the USA, tazarotene is also licensed for use in the treatment of acne and several studies show its efficacy for the treatment of other skin diseases like photodamage and ichthyoses. PMID- 14748201 TI - [Medication of the month. Evra: first contraceptive transdermal patch]. AB - Evra is a transdermal patch releasing 20 micrograms of ethinylestradiol and 150 micrograms of norelgestromin/day during one week. The circulating levels of steroids attained are of similar amplitude though steadier than after intake of an oral combined low-dose estrogen-progestin pill. The transdermal method is user friendly and is abided by a high degree of acceptability, and a low level of skin irritability. Its contraceptive effectiveness is similar to that conferred by oral contraceptives except if the treated woman is over 90 kg, in which case Evra should not be prescribed. Cycle control is excellent and similar to that of triphasic pills. Adverse effects and tolerance are comparable to those described with low-dose oral contraceptives with a slight estrogen dominance. Lipid and glucose metabolism as well as coagulation are influenced in the same way. Gastrointestinal disturbances (nausea, vomiting) do not prevent the efficacy of the transdermal patch. Compliance with Evra is significantly higher than with oral combined contraceptives--a major point for an effective contraception. Accordingly, Evra constitutes an useful addition to the current array of contraceptive methods. PMID- 14748202 TI - [Clinical study of the month. The EUROPA study: cardiovascular protection with perindopril in patients with stable coronary heart disease]. AB - The multicentre placebo-controlled double-blind "EUropean trial on Reduction Of cardiac events with Perindopril in patients with stable coronary Artery disease" (EUROPA) assessed whether the angiotensin-converting-enzyme inhibitor perindopril reduces cardiovascular risk in a population with stable coronary heart disease and no apparent heart failure, whatever the associated cardiovascular risk. Patients were randomly assigned perindopril 8 mg once daily (n = 6110) or matching placebo (n = 6108). After a mean follow-up of 4.2 years, a relative risk reduction of 20% (95% CI 9-29, p = 0.0003) was observed in the combined primary endpoint (cardiovascular death, myocardial infarction, or cardiac arrest) in the group treated with perindopril as compared to placebo. About 50 patients needed to be treated for a period of 4 years to prevent one major cardiovascular event. This benefit was consistent in all predefined subgroups. According to these results, treatment with perindopril, on top of other preventive medications, should be considered in all patients with stable coronary heart disease. PMID- 14748203 TI - Neuroimaging in autistic spectrum disorder (ASD). AB - Autistic spectrum disorder (ASD) is a lifelong developmental disorder characterized by impairment in socialization and communication. Neuroimaging research has shown abnormalities in the frontal lobes, limbic systems, and cerebella of individuals with ASD. Recently, abnormal developmental trajectories of brain growth have been reported, with increases in brain volume (in both gray and white matter) seen in younger rather than older individuals with this disorder. Despite 30 years of research, a reliable marker for ASD has not been identified. Therefore, routine neuroimaging for individuals with ASD is not recommended. PMID- 14748204 TI - Characteristics and predictors of aortic plaques in patients with transient ischemic attacks and strokes. AB - OBJECTIVE: To identify the prevalence and characteristics of aortic atherosclerotic plaque disease and its association with cerebrovascular risk factors in patients with cerebral ischemic events. BACKGROUND: Aortic atheroma is associated with ischemic stroke. Its characteristics, including morphology and distribution among different stroke subtypes, are not well described. METHOD: From July 2000 to August 2001, all patients evaluated by transesophageal echocardiography (TEE) with diagnoses of transient ischemic attacks (TIAs) and strokes were prospectively studied. Demographics, including age, gender, ethnicity, cerebrovascular risk factors, and stroke subtypes, were collected. RESULTS: Thoracic aortic atheromas (TAAs) were present in 141 of 237 patients (59%) (mean age = 59 +/- 14, 119 [50%] male). Mild plaque (< 2 mm) was present in 13 of 237 (5%), moderate plaque (2-4 mm) in 49 (21%), severe plaque (> or = 4 mm) in 79 (33%), and complex plaque in 64 (27%). Patients' ages (odds ratio [OR] = 1.05, confidence interval [CI] 1.03-1.08, P < .001), coronary artery disease (OR = 2.2, CI 1.02-4.8, P < .042), and patent foramen ovale (PFO) (OR = 0.39, CI 0.22 0.70, P < .002) were associated with the severity and complexity of aortic plaque. In multivariate analysis, age (OR = 1.06, CI 1.03-1.08, P < .001) and the presence of PFO (OR = 0.35, CI 0.18-0.65, P < .001) continued to be significant to the severity and complexity of aortic atheroma. Gender, history of stroke, hypertension, diabetes mellitus, hyperlipidemia, and history of smoking were not associated with TAA. CONCLUSION: One third of TAA plaques are severe and complex in nature and more frequently present in the descending aorta and the arch of the aorta than in the ascending aorta. TEE should be considered for the early detection and treatment of TAA in patients without identified causes of stroke. PMID- 14748205 TI - Voxel-based mapping of cortical ischemic damage using Tc 99m L,L-ethyl cysteinate dimer SPECT in acute stroke. AB - BACKGROUND AND PURPOSE: When performed soon after stroke onset, single-photon emission computed tomography (SPFCT) with hexamethylpropylenamine oxime or Tc 99m L,L-ethyl cysteinate dimer (ECD) has significant added predictive value compared to neurological scores. With ECD SPECT, the degree of tracer uptake reduction predicts neurological recovery, and using a 40% threshold to characterize irreversibly damaged tissue (IDT), significant correlations have been observed. However, correlations between this uptake threshold and tissue outcomes have not been assessed. The purpose of this study was to validate the 40% ECD uptake threshold for the probabilistic mapping of IDT using an automatic, voxel-based approach. METHODS: In 10 acute stroke patients, the authors first compared early ECD SPECT and late coregistered magnetic resonance imaging (MRI) data and assessed for IDT and "tissue at risk" (i.e., tissue with ECD uptake below and above 40%) the percentage of voxels ultimately infarcted and noninfarcted on late brain MRI. They then assessed the correlations between the volumes of brain tissue compartments and subsequent neurological recovery. Finally, to assess whether visual SPECT analysis is reliable compared to the more complex voxel based approach, the authors compared the predictive value of the 2 methods for neurological recovery. RESULTS: The majority of IDT voxels (average = 84%), defined by ECD uptake < 40%, evolved toward infarction, and 51.8% to 100% of at risk voxels (average = 89%) escaped infarction. The extent of IDT correlated significantly with neurological recovery (P = .0009). There was good agreement between visual and voxel-based analyses (P = .0004). CONCLUSION: The results support the validity of the ECD uptake thresholds chosen, suggesting that ECD uptake can reflect neuronal viability and that ECD SPECT can be useful for the early detection of potentially salvageable tissue and irreversible damage. These preliminary results encourage the use of this method in a clinical setting for fast decision making in choosing acute therapy. PMID- 14748206 TI - Neuroimaging of the complications of epilepsy surgery. AB - BACKGROUND AND PURPOSE: Stroke-like symptoms can be associated with the invasive evaluation and surgical resection of epileptic foci in patients with intractable epilepsy. Neurological deficits following surgical procedures for epilepsy are not uncommon, but most are relatively minor and transient. The authors investigated the neuroimaging patterns of cerebral tissue insults in patients suffering neurological deficits directly related to procedures performed to evaluate and treat intractable epilepsy. They attempted to discern potential secondary vascular insults from the not unexpected tissue loss that can be associated with various epilepsy procedures. METHODS: The authors prospectively assessed 7 consecutive patients who underwent either the invasive electrocortigraphic monitoring or surgical resection of epileptic foci. All had some degree of neurological deficit postoperatively. The authors evaluated for tissue injury type with postoperative computed tomography and magnetic resonance brain imaging. They also review pertinent medical literature addressing potential complications of epilepsy surgery. RESULTS: Three patients had primarily ischemic tissue injuries, 2 had tissue loss with minor bleeding, and 1 transient deficit appeared to reflect the amount of tissue removed. Another patient had choreiform movements and gait ataxia 1 week after the procedure, but no follow-up neuroimaging was available. The primary ischemic insults appeared to be related to vascular traction or compression or possibly vasospasm. The tissue loss/hemorrhagic insults were presumably related to tissue loss, with seepage of blood or bleeding from a resected cavernous hemangioma. No patients died, but 1 was left with a persistent, moderately severe neurological deficit. CONCLUSIONS: It is important to distinguish the not unexpected neurological deficits associated with inadvertent trauma to normal brain tissue during procedures associated with epilepsy surgery from vascular insults. Postoperative neuroimaging can be useful in this endeavor. PMID- 14748207 TI - Brain tissue water uptake after middle cerebral artery occlusion assessed with CT. AB - BACKGROUND AND PURPOSE: To study whether computed tomography (CT) can measure the water content of early ischemic edema. METHODS: The authors obtained cranial CT in 5 groups of rats subjected to 1 hour (n = 8), 2 hours (n = 11), 3 hours (n = 13), 4 hours (n = 13), or 6 hours (n = 14) of right middle cerebral artery (MCA) occlusion. Immediately after CT, the authors removed the rats' brains and determined tissue water content by the dry-wet weight method. They correlated brain x-ray attenuation with brain tissue water content. RESULTS: Mean brain tissue water content remained constant in the nonischemic left hemispheres at 77.9% +/- 0.6% and increased up to 79.3% +/- 1.0% in the right hemispheres after 6 hours of permanent right MCA occlusion. X-ray attenuation remained constant in the left hemispheres at 75.6 + 2.2 Hounsfield units (HU) and decreased to 71.7 +/ 3.4 HU in the right hemispheres after 6 hours of right MCA occlusion. The decrease in x-ray attenuation correlated significantly with the increase in ischemic brain tissue water content (y = 217.3 - 1.8x x; r = .55, P < .0001). That means that a 1% increase in hemispheric tissue water content causes a decrease in x-ray attenuation of 1.8 HU. CONCLUSIONS: After MCA occlusion, immediate brain tissue net water uptake is associated with a decrease in x-ray attenuation. CT can monitor ischemic edema in an acute stroke. PMID- 14748208 TI - Cortical reorganization allows for motor recovery after crossed cerebrocerebellar atrophy. AB - The authors report the case of a 33-year-old woman who exhibited, at the age of 17, a left-sided hemiplegia, which was followed by good motor recovery, though with a permanent deficit in fine finger movements. She had a widespread loss of neural tissue in the right hemisphere (crossed cerebrocerebellar atrophy), including (1) marked atrophy and thinning of the precentral and postcentral gyri; (2) widespread deep white matter destruction, including the corticospinal tract; and (3) crossed cerebellar atrophy. Except over the supplementary motor area (SMA), transcranial magnetic stimulation did not elicit motor evoked potentials in the affected hand. Nevertheless, during opening and closing of the affected hand, functional magnetic resonance imaging showed an activation of the lesioned primary sensorimotor cortex (SMC), as well as of the intact SMA and the parietal areas, but not of the ipsilateral motor areas. The authors speculate that recovery was achieved by a motor command generated in the SMC and the parietal cortex, passing through corticospinal axons originating in the SMA. PMID- 14748209 TI - Multiple cerebral microbleeds: MRI marker of a diffuse hemorrhage-prone state. AB - BACKGROUND: Recent reports have indicated that cerebral microbleeds (CMBs) detected on gradient-echo (GRF) magnetic resonance imaging may be a risk factor for therapy related intracranial hemorrhages (ICHs). CASE DESCRIPTION: The authors describe 3 patients with multiple CMBs in whom ICHs occurred either after tissue plasminogen activator administration or after the initiation of antiplatelet therapy. Hemorrhages occurred in brain areas with normal appearances on GRE imaging and remote from the CMBs. CONCLUSION: Multiple CMBs may signal a diffuse hemorrhage-prone vasculopathy. PMID- 14748210 TI - Treatment of Marburg variant multiple sclerosis with mitoxantrone. AB - Marburg variant multiple sclerosis (MS) is a severe, sometimes monophasic, form of MS leading to advanced disability or death within a period of weeks to months. No consistently successful treatment for Marburg variant MS has been described. The case reported here is that of a 34-year-old woman with Marburg variant MS whose magnetic resonance imaging scan showed extensive brainstem, periventricular white matter, subcortical, and cortical involvement, giving the appearance of an "MS cerebritis." The patient had no response to treatment with methylprednisolone but improved and recovered after treatment with mitoxantrone (MITX). Because MITX has potent anti-inflammatory effects and demonstrated efficacy in worsening MS, it may be an appropriate agent for the treatment of Marburg variant MS. PMID- 14748211 TI - Heidenhain variant of Creutzfeldt-Jakob disease: diffusion-weighted MRI and PET characteristics. AB - Creutzfeldt-Jakob disease (CJD) is characterized by rapidly progressive dementia with a variety of neurological disorders and a fatal outcome. The authors present a case with visual disturbance as a leading symptom and rapid deterioration in global cognitive functions. The cerebrospinal fluid was positive for 14-3-3 protein, and diffusion-weighted magnetic resonance imaging (MRI) showed marked hyperintensity in the parieto-occipital cortices, where hypometabolism was clearly detected on positron emission tomography (PET). Pattern-reversal visual evoked potentials showed prolonged P100 latencies and increased N/5/P100 amplitudes. All these findings supported a diagnosis of the Heidenhain variant of CJD, whereas a long clinical course, a lack of myoclonus, and an absence of periodic synchronous discharges on electroencephalography were atypical. Diffusion-weighted MRI and PE1 in combination with visual evoked potential recording and 14-3-3 protein detection may be useful for the early diagnosis of CJD. PMID- 14748212 TI - Aberrant ascending pharyngeal artery mimicking a partially occluded internal carotid artery. AB - Doppler sonography has become a primary imaging modality for the diagnosis of carotid arterial stenosis. Carotid stenting for a severely stenotic but not completely occluded carotid artery is becoming an alternative to carotid endarterectomy in selected groups of patients. The authors discuss a case of complete occlusion of the internal carotid artery associated with an ipsilateral aberrant ascending pharyngeal artery originating from the proximal internal carotid artery, which mimicked a stenotic internal carotid artery on sonography. Meticulous Doppler sonographic examination may provide clues for this extraordinary condition, yet angiography is indicated for a definite diagnosis. PMID- 14748213 TI - Cerebral metastasis from heart angiosarcoma presenting as multiple hematomas. AB - The authors present the case of a 24-year-old woman with cerebral metastasis from a primary heart angiosarcoma, which appeared as multiple cerebral hematomas. Primary or metastatic brain angiosarcomas are exceedingly rare, and only a few cases have been reported with hemorrhage. Initial neurological symptoms were mild hemiparesis with numbness, and chest pain was first misdiagnosed as pericarditis. A computed tomography (CT) scan and magnetic resonance imaging showed 2 hematomas in the left parietal and occipital lobes. A thoracic CT scan revealed angiosarcoma of the right atrium of the heart with multiple infiltrations of the lung. The patient underwent surgical removal and systemic chemotherapy. She died 6 months after surgery. PMID- 14748214 TI - Christian-based counseling for the suicide survivor: a guide for pastoral therapy. AB - This article addresses the history of stigmatization experienced by suicide survivors, specifically through biblical and theological thought evolving from the Christian Church. Contemporary understandings and attitudes are then assimilated. Four grief stages specific to the grief experience of the suicide survivor are introduced--anger, guilt, shame, and loss--with suggested approaches at each state intended to aid the pastoral counselor in better understanding and ministering to the suicide survivor. A summary of a recommended congregational response is also provided. PMID- 14748215 TI - Pastoral counseling among Chinese churches: a Canadian study. AB - This article reports the results of a survey of Canadian Chinese pastors (N 20) regarding their experiences and attitudes related to pastoral counseling. It identifies common concerns encountered during pastoral counseling, as well as key areas of church concerns. Results of the survey indicate that pastoral counseling may be more important than is currently recognized by Chinese pastors, particularly in key areas of marriage, family life, and general relationship issues. The authors discuss possible implications of the survey relative to training and equipping Chinese pastors to engage more in pastoral counseling as an integral part of their parish ministry. PMID- 14748216 TI - Homecoming rituals: weaving multicultural funeral narratives. AB - Using the personal experience of "homecoming" funcrary journey from the U.S. to Korea the author of this article probes the cultural construction of familial bond in Korean and biblical (Genesis) tuneral narratives. He highlights the cultural differences between American and Korean funeral traditions--(1) dying at hospital vs. dying at home, (2) embalming vs sup/yom (washing/binding of the corpse), (3) metal casket vs. wooden coffin, (4) committal vs. banhon (returning of the spirit)--to explore the religious and practical implications of the multicultural grieving process for pastoral care and grief ministry in which death is conceived as returning to the familial, cultural, and spiritual homes. PMID- 14748218 TI - Informed consent and parish clergy. AB - Pastors are given permission by others to extend care and to practice ministry. This permission may be given in denominational policy, congregational expectations, or individual requests for pastoral care. At the individual level, permission for pastoral care can be understood as analogous to the notion of informed consent in health care ethics. Informed consent is a compound concept involving: (1) the sharing and clarifying of relevant information, and (2) the respect for a person's consent for care or refusal of care. It functions as a guard both against harm and against unwanted beneficence. Because many expectations for pastoral care are not explicitly stated, ministers need to develop sensitivity to hear both requests for help and requests for privacy. PMID- 14748217 TI - "Worlds apart": truth telling in the case of Mrs. VV. AB - This article discussed how the views of a terminal cancer patient and her family, who were immigrants from Romania, differed from those of her North American caregivers. It presents evidence about truth telling to cancer patients in Eastern Europe and general cultural differences in attitudes to truth telling. Literature on how to span the gap between different cultural worlds is reviewed. Conclusions address ways to minister to patients who come from "different worlds" than their caregivers. PMID- 14748219 TI - Spiritual pain: a brief overview and an initial response within the Christian tradition. AB - The author notes that spiritual pain is widespread, both within individuals and society. Drawing on personal experiences and a literature survey, he offers an overview of current work on spiritual pain. As distinct from palliative caregivers, he suggests that spiritual pain can be noticed, but is difficult to define. Drawing on some of the Church's spiritual masters, the author offers a broad typology for spiritual pain, claiming that current methods for attending to spiritual pain are too complex to be useful. He therefore offers, and briefly explores, the image of retreat for attending to spiritual pain--compassionate hospitality, tempered by a form of "tough love" that enables the retreatAnt to embrace, welcome, and dialogue with pain. He suggests that essential features within such an image include an experience of support; a safe and sacred space, a sense of freedom; appropriate expressions of reassurance, and opportunities for forgiveness and reconciliation. PMID- 14748220 TI - Self-mutilation: a spiritual endeavor. AB - This article explores the phenomenon of self-mutilatory behaviors among practitioners both today and throughout history with the intent of exposing what the author finds to be a radical correlation between mental illness and spirituality. It is the author's belief that today's strong focus on the biology of mental illness, though essential, will extend us only so far in our comprehension of mental pathology; and that an investigation into the spiritual needs of humankind may serve to round out our understanding. PMID- 14748221 TI - Need more than unsubstantiated claims regarding Jewish supervisees in CPE. PMID- 14748222 TI - Pastoral care/pastoral counseling. More similarities than differences: who does it serve and show us the evidence. PMID- 14748223 TI - Perspectives. Some tough nuts to crack in first AHRQ quality report. PMID- 14748224 TI - Taking the next step forward for ICD-10. PMID- 14748225 TI - The drive for an EHR standard picks up speed. PMID- 14748226 TI - Your guide to implementing the EHR. AHIMA work groups deliver best practices. PMID- 14748227 TI - Taking electronic document management beyond the HIM department. PMID- 14748228 TI - Pioneers and perseverance. Implementing the EHR in physician practices. PMID- 14748229 TI - Apples to apples. Using autobenchmarking to measure productivity. PMID- 14748230 TI - HIPAA and the EHR: making technical safeguard changes. PMID- 14748231 TI - Developing E-health standards for Web sites. PMID- 14748232 TI - Building an enterprise master person index. PMID- 14748233 TI - Finding quality HIPAA security resources. PMID- 14748234 TI - Evaluating compliance program effectiveness. PMID- 14748235 TI - Endoscopic surgery brings relief to sinus patients. PMID- 14748236 TI - Changes abundant for CPT 2004. PMID- 14748237 TI - Making patient safety number one. PMID- 14748239 TI - CPT: what's in store for 2004? PMID- 14748238 TI - Two ways of looking at a healthy practice. PMID- 14748240 TI - Medicare payments to increase as part of Rx drug bill. PMID- 14748241 TI - Five strategies for a more vital practice. PMID- 14748242 TI - Creating a successful after-hours clinic. PMID- 14748243 TI - Three steps to an effective practice budget. PMID- 14748244 TI - Weighing the risks and benefits of clinical interventions. PMID- 14748245 TI - Creating an electronic filing cabinet. PMID- 14748246 TI - CMS' pay-per-visit idea for nephrologists is a reasonable one--let's see if it improves patient outcomes. PMID- 14748247 TI - Improving retention and motivation in non-clinical dialysis employees. AB - For no additional expense, little intrusion into the schedule or duties of the day, and no additional personnel to hire, dialysis facilities can make a dramatic impact on the training, motivation, and retention of employees through the use of classes designed specifically for the needs of newly employed non-clinical personnel. In today's world where many are expected to do a task with little or no orientation to the overall values and goals of the organization and how their tasks matter, what other action can accomplish so much for employee motivation and retention for so little time and expense? At DCI Mid-Missouri this program has been successful in retaining many excellent employees over many years and has been part of an ongoing effort to increase employees' interest and commitment to their work and the organization. It has also made them much more aware of treatments, other personnel and most of all, of our patients and their needs. It fulfills the needs of humans to be valued and have meaningful work. It contains costs and helps efficiency and productivity. Most of all, it keeps excellent people on the job and enjoying their work more than they would have. Participants' words when evaluating the most recent series of classes speak for themselves in demonstrating these important benefits that can be achieved easily in dialysis facilities throughout the United States and the world. PMID- 14748248 TI - Chronically motivated. Hartwell works 24/7 to improve the patient-caregiver relationship. PMID- 14748249 TI - Three ways to protect the assets of your nephrology practice. PMID- 14748250 TI - Healing the body and the soul of the CKD/ESRD patient. PMID- 14748251 TI - Modernizing local responses to public health emergencies: bioterrorism, epidemics, and the model state emergency health powers act. PMID- 14748252 TI - HIV and art: reproductive choices and challenges. PMID- 14748253 TI - Averting the clone age: prospects and perils of human developmental manipulation. PMID- 14748254 TI - Turning a blind (white) eye in legislating mental health parity: the unmet, overlooked needs of the working poor in racial and ethnic minority communities. PMID- 14748255 TI - Recent developments in direct consumer advertising of attention disorder stimulants and creating limits to withstand constitutional scrutiny. PMID- 14748256 TI - Pregnant drug abusers are treated like criminals or not treated at all: a third option proposed. PMID- 14748257 TI - Should your ERISA remedy depend upon your geography?: an analysis of Rush Prudential HMO, INC. v. Moran. PMID- 14748258 TI - [Macrolide antibiotic resistance rates and phenotypes of group A beta hemolytic streptococci isolated between the years 1999-2000 and 2001-2002]. AB - Macrolide resistance in group A beta hemolytic streptococci (GABHS) have been reported with increasing frequency from various geographic regions in the world, and for the respective treatment alternatives and epidemiologic studies, macrolide resistance rates and phenotypes have been determined. In this study erythromycin, clarithromycin and azithromycin were tested with disc diffusion method against 560 GABHS, isolated from the throat samples collected between 1999 2002. NCCLS guidelines were followed for the susceptibility tests and MIC values were obtained by E-Test in resistant isolates. For determining the resistance phenotype, erythromycin and clindamycin discs were used. Only one isolate (0.27%) was found intermediately resistant to erythromycin, and clarithromycin, and resistant to azithromycin between May 1999-January 2000, whereas in the period between January 2001-June 2002, one isolate (%0.5) was found susceptible to erythromycin and clarithromycin, but intermediately resistant to azithromycin. In each period three isolates (0.83% and 1.5%, respectively) were found to be resistant to all the tested macrolides. There was no statistically significant difference between the resistance rates in these periods. Three of the resistant isolates had inducible type, and the other five isolates had M phenotype macrolide resistance. Testing for macrolide susceptibilities and screening the resistance phenotypes have crucial importance in case of GABHS infections, since these can be taken into consideration for epidemiological issues as well as a guide for empirical treatment protocols in any geographical setting, as well as in our country. PMID- 14748259 TI - [Sensitivity of methicillin resistant staphylococci to linezolid and some other antimicrobial agents]. AB - Linezolid is a synthetic antimicrobial agent which was introduced into clinical therapy in the early 2001. It has an inhibitory effect against most of the Gram positive bacteria, including methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin resistant enterococci. In this study, in-vitro activities of linezolid, teicoplanin, vancomycin, ciprofloxacin, gentamicin, erythromycin, clindamycin, trimethoprim/sulfamethoxazole and rifampicin, were tested against 164 methicillin-resistant Staphylococcus spp. (96 S. aureus and 68 coagulase negative staphylococci) isolated from clinical specimens, by using disk diffusion method. None of the strains were found to be resistant to linezolid, vancomycin and teicoplanin. The resistance rates to other drugs were as follows; 80.5% to ciprofloxacin, 78.0% to gentamicin, 76.8% to erythromycin, 65.5% to clindamycin, 57.3% to trimethoprim/sulfamethoxazole and 41.5% to rifampicin. It was concluded that linezolid can be used as an alternative drug in severe infections caused by methicillin-resistant Staphylococcus spp., especially if the isolate was found to be resistant to teicoplanin or a side effect of vancomycin was observed. PMID- 14748260 TI - [Antibiotic resistance of Acinetobacter baumannii isolates: data from Ibni Sina Hospital for the year 2002]. AB - Acinetobacter spp. have a special role in nosocomial infections. They usually colonize intensive care units and cause infections. Multiple antibiotic resistance is observed to be an important problem in Acinetobacter isolates in recent years. The aim of this study was to evaluate the susceptibilities of 277 Acinetobacter baumannii strains (99 blood, 71 abscess, 26 urine, 81 lower respiratory tract isolates) isolated from adult patients hospitalized in Ankara University Ibni Sina Hospital during one year period in 2002. The resistance rates of the strains against studied antibiotics were found as follows; 31.2% for netilmicin, 44.6% for sulbactam-cefoperazone, 53.6% for imipenem, 59.8% for amikacin, 59.9% for tobramycin, 74% for ciprofloxacin, 78% for gentamicin, 78.3% for ticarcillinclavulanate. 79.5% sulbactam-ampicillin, 82.3% for cotrimoxazole, 84.8% for ticarcillin, 87.3% for piperacillin-tazobactam, 88.1% for ceftazidime and 92.1% for piperacillin. When the isolates obtained from the reanimation unit were compared with the isolates obtained from the other hospital units, a significantly high level of resistance was found from the isolates obtained from the reanimation unit except netilmicin and sulbactam/cefoperazone. The high rate of in-vitro antibiotic resistance of the A. baumannii strains indicated the importance of controlled antibiotic usage and appliance of hospital infection control measures. PMID- 14748261 TI - [Seropositivity against atypical pneumonia agents demonstrated in patients with community-acquired pneumonia]. AB - The aim of this study was to investigate the IgG and IgM antibody positivities against atypical pneumonia agents in patients with community-acquired pneumonia (CAP), and to compare the results with the controls. The serum samples which were collected from 53 adult patients and 20 healthy donors have been investigated by a commercial indirect immunofluorescent assay (IFA, Pneumo-slide, Vircell SL, Spain) in which nine different antigens were fixed onto a slide. In both of the study groups. IgG and IgM seropositivities were detected in different rates against one or more etiologic agents. In the patient group. IgG and IgM positivity rates for the agents were as follows, respectively; 22.6% and 28.3% for Legionella pneumophila, 9.4% and 5.6% for Mycoplasma pneumoniae, 30.1% and 7.5% for Coxiella burnetii, 33.9% and 3.7% for Chlamydia pneumoniae, 28.3% and 0 for adenovirus, 71.6% and 1.8% for respiratory syncytial virus, 30.1% and 24.5% for influenza A virus, 35.8% and 7.5% for influenza B virus, 71.6% and 1.8% for parainfluenza viruses type 1-3. The rates of IgG positivities in the control group varied between 5-55% for all of the agents except M. pneumoniae and 3 of these controls were positive for L. pneumophila IgM, 3 were positive for C. pneumoniae IgM and one was positive for influenza A virus IgM. According to the statistical evaluation, there were no significant differences for IgM seropositivities to any of the agents, between the patient and control groups (p > 0.05). These results could be attributed to one or more of the following; a) none of these microorganisms were the primary etiologic agents, b) IgM positivities were the result of reinfections with these agents, c) longer duration of IgM antibodies after the acute infections. In terms of IgG positivities between the patient and control groups, only C. burnetii showed statistically significant difference (p = 0.029). Since the type of the pathogens causing CAP are of crucial importance both for the epidemiological purposes and for planning the empirical treatment strategies, more detailed multicenter studies should be performed in our country. PMID- 14748262 TI - [Clinical characteristics of Lyme disease in 12 cases]. AB - Lyme disease, an infection caused by Borrelia burgdorferi, has been reported in many countries from America and Europe, however, knowledge about its epidemiology in Turkey is incomplete. In this study, the clinical characteristics of 12 cases with Lyme disease who were diagnosed with the positivity of B. burgdorferi antibodies by western blot method in the laboratory of Ankara Bayindir Medical Center, have been reviewed. Physicians' recognition of early symptoms such as erythema migrans and later findings pertaining to the nervous system, joints, eye, and skin, and general awareness of the role of tick bites may increase the rate of diagnosis and allow earlier treatment of Lyme disease. PMID- 14748263 TI - [The value of the immunoglobulin G avidity test for the serologic diagnosis of brucellosis]. AB - In order to investigate the value of immunoglobulin G (IgG) avidity test for the serological diagnosis of Brucella infections, a total of 118 patients (74 male, 44 female; mean age: 43 +/- 18.7 years) were included into the present study. The patients have been diagnosed with the characteristic clinical findings, > or = 1/160 antibody titers in standard tube agglutination (STA) test and/or blood culture positivities. Brucella spp. have been grown in blood cultures of 78 patients, and STA test results were found positive (> or = 1/160 titers) in 117 patients. The diagnosis of a patient with 1/80 STA titer was based on the blood culture positivity. By enzyme immunoassay (ELISA), 3 patients (2.5%) were found positive for IgM, 14 (11.9%) were positive for IgG, and 101 (85.6%) were positive for both IgM and IgG. The patients who were found IgG positive have been grouped according to their duration of complaints. Group 1 included 99 patients with the history of brucellosis < or = 6 months, and group 2 included 16 patients with the history of brucellosis > 6 months. IgG avidity test was performed by ELISA in 115 IgG positive serum samples, with the denaturation substance (8 M urea). The cut off value for IgG avidity index (AI) was accepted as 40%, and the avidity maturation period was defined as 6 months. As a result, the rates of patients who had low IgG AI in group 1 and 2 were found as 91.9% and 43.7%, respectively, while these rates were 8.1% and 56.3% for the presence of high IgG AI, respectively. The rate of low AI in group 1 was an expected result, while the rate was more than expected in group 2, indicating that Brucella antibodies with low avidity indices would not be helpful for the diagnosis of a recent infection, while Brucella antibodies with high avidity indices would be useful for the elimination of a recent infection. PMID- 14748264 TI - [Comparison of different methods for the identification of Candida species isolated from clinical specimens]. AB - The aim of this study was to compare the different methods for the identification of Candida strains isolated from clinical specimens. The methods of germ tube examination, chlamydospore examination formed on the rice Tween-80 (RT-80) agar and evaluation of colony morphologies on the two chromogenic agars (CHROMagar Candida, Albicans ID), were compared with a reference API 20C AUX (bioMerieux, France) automated system based on the carbohydrate assimilation, for the identification of a total 255 Candida isolates. Of them, 173 (67.8%) were identified as C. albicans, 37 (14.5%) were C. glabrata, 23 (9%) were C. krusei, 9 (3.5%) were C. tropicalis, 9 (3.5%) were C. kefyr, 2 (0.8%) were C. guillermondii and 2 (0.8%) were C. parapsilosis, by API 20C AUX system. In the view of these results, 146 (84.4%) of C. albicans strains were identified by germ tube examination, 161 (93.1%) of C. albicans strains and 208 (81.5%) of total strains were identified by chlamydospore examination. 169 (97.7%) of C. albicans strains and 231 (90.6%) of total strains were identified by CHROMagar Candida method, and 168 (97.1%) of C. albicans strains were identified by Albicans ID method, correctly. In the CHROMagar Candida medium, 169 C. albicans isolates have produced bright green colored colonies, whereas 33 (89.2%) isolates which produced dark pink/purple colored colonies were identified as C. glabrata, 7 (77.8%) isolates which produced metalical blue colored colonies were identified as C. tropicalis and 22 (95.6%) isolates which produced pale pink colored colonies were identified as C. krusei. In the Albicans ID medium, four of the 172 isolates which were evaluated as C. albicans initially by producing blue colored colonies, have been identified as C. tropicalis by API 20C AUX system. The sensitivities and specificities of germ tube examination, RT-80, CHROMagar Candida and Albicans ID methods were found as follows, respectively; 84.4% and 100%, 93.1% and 100%, 97.7% and 100%, 99.4% and 95.3 percent. In conclusion, CHROMagar Candida medium seems the most favorable rapid and practical method with high sensitivity and specificity for the identification of Candida species, but its cost-effectiveness should be kept in view. PMID- 14748265 TI - [The use of real-time polymerase chain reaction and enzyme immunoassay for the diagnosis of acute parvovirus B19 infections in immunosuppressed patients]. AB - Human parvovirus B19 (PV-B19) infection may lead to very serious clinical situations such as transient aplastic crisis in patients with hemolytic anemia, thrombocytopenia, neutropenia and transient arthritis accompanied with erythema infectiosum, especially in immunosuppressed patients. Early diagnosis of PV-B19 infection is of critical importance especially in immunosuppressed patients since the necessary precautions can be undertaken accordingly. In this study, PV-B19 IgM and IgG antibodies and viral DNA have been searched by enzyme immunoassay (ELISA) and real-time polymerase chain reaction (PCR), respectively, in 50 PV-B19 suspected immunosuppressed patients. Viral IgM, IgG and DNA positivities were detected in 7 (14%), 20 (40%) and 7 (14%) of the patients, respectively. During the first week three patients were found DNA and IgM positive but IgG negative, while four patients were found positive for the viral DNA, IgM and IgGs. The DNA copy numbers were high in all of the patients during the first week, with a gradual decrease during a seven-week follow-up period. IgM antibodies have disappeared in the sixth week in three of the patients and at the end of the seventh week in four of the patients. Although the IgG antibodies were negative in three patients in the first week, they became positive in the second week and the titers gradually increased during the following weeks. According to the results of this study, it can be concluded that, in high risk groups such as immunosuppressed patients, in addition to ELISA, real-time PCR method would be helpful for the early diagnosis of PV-B19 infections. PMID- 14748266 TI - [Seroprevalence of hepatitis B and C infections in surgical personnel and evaluation of predisposing factors]. AB - The aim of this study was to investigate the seroprevalence of hepatitis B and C infections in the personnel of the operating room of Ondokuz Mayis University Hospital, to detect the carriers and actively infected individuals, to find out the health-workers' behaviours for the prevention from these infections, and to ascertain the risk factors of these diseases. HBsAg, anti-HBc, anti-HBs and anti HCV antibodies were examined in the serum samples of 190 personnel, and their gender, education, working time (year), the daily working time in operating theatre (part/full time), the history of past hepatitis B infection, hepatitis B immunization, history of a past operation and blood transfusion, and the use of gloves during working (continuously-intermittent) were questioned. As a result, no carrier and/or actively infected personnel with hepatitis B and C were detected, while 36 (18.9%) of them had a history of past hepatitis B infection, and 132 (69.5%) of them had hepatitis B vaccine. The continuous glove use was more common in part-time workers than those of full-time workers, the history of past hepatitis B infection was increasing as the duration of working years increase, and the rates of being vaccinated against hepatitis B was decreasing as the duration of working years increase (p < 0.05). PMID- 14748267 TI - [A case of acute pancreatitis due to Weil's disease]. AB - Leptospirosis, the most common vasculitic zoonosis in the world, is characterized with jaundice and acute renal failure. However, pancreatitis is an uncommon complication of leptospirosis. In this report, an acute pancreatitis case due to Weil's disease has been presented. A 31-year-old female patient with high levels of glucose, blood urea nitrogen, creatinine, creatine kinase, bilirubin, amylase and lipase, has been diagnosed to have leptospirosis by the high positive result (1/800) of microscopic agglutination test against Leptospira interrogans serogroup icterohemorrhagiae. The patient has been treated with supportive and symptomatic therapy, and with penicillin G for leptospirosis. Following triple hemodialysis, all the blood biochemistry tests returned to normal on the tenth day of therapy. This case was reported to draw attention to Leptospira infections which should be considered in the differential diagnosis of patients with jaundice and pancreatitis. PMID- 14748268 TI - [Prolonged hepatitis A: three case reports]. AB - In this report, three patients, admitted to Ankara University Faculty of Medicine, Ibni Sina Hospital, Clinical Bacteriology and Infectious Diseases Department, with the diagnosis of acute viral hepatitis A have been discussed because of their clinical and/or biochemical relapses. Following conservative treatment and bedrest, the liver transaminases decrease to nearly normal levels but a second peak was observed later in three of the cases. Other viral, metabolic and autoimmune factors which could lead to the increase of liver transaminases were ruled out. Anti-HAV IgM antibodies became negative in 2 to 4 months later, and liver enzymes returned to normal in 4 to 5 months after onset of their illnesses. As a result, it was concluded that conservative follow-up and therapy would be sufficient since the prognosis of A hepatitis is usually benign even if it relapsed. PMID- 14748269 TI - [Streptococcal toxic shock syndrome: a case report]. AB - Streptococcal toxic shock syndrome (STSS) is the most severe form of invasive infections caused by group A streptococci. In this report, a 36-years-old man who was admitted to our clinic with the complaints of fever, rash, skin lesions, abdominal pain, weakness and anuria for 2 days, has been presented. His body temperature was 39.5 degrees C and blood pressure was 50/20 mmHg. In physical examination, diffuse erythematous rash on the body, cellulitis on left leg and foot, fungal lesions on the toes, and abdominal tenderness were noted. Laboratory results revealed a dramatic increase in leukocyte count, increased sedimentation rate, elevated blood urea nitrogen, cretinine, liver enzymes and bilirubin levels. Group A streptococci were isolated from the blood culture of the patient. Despite supportive (intravenous saline, dopamine) and antibiotic (clindamycin ceftriaxone combination) therapies, adult respiratory distress syndrome has developed in two days, and he died on the third day. This case was presented to draw attention to STSS, which was a rare clinical entity with rapid progression to mortality despite aggressive medical therapy. PMID- 14748270 TI - [A new marker for the diagnosis of rheumatoid arthritis: cyclic citrullinated peptide antibodies (anti-CCP)]. AB - Rheumatoid arthritis (RA) is one of the most common autoimmune diseases, affecting about 1% of the world population. Rapid diagnosis and early treatment are important to control the progression of the disease. The most commonly used test for the serological diagnosis of suspected RA cases is the determination of rheumatoid factor (RF). RF is a sensitive but not very specific marker for RA. Of 40-60% RA patients also exhibit autoantibodies against epidermal flaggrin (RA keratin, anti-perinuclear antibody) in the serum. In the recent years, it has been shown that a rarely found amino acid citrulline, which is present in flaggrin, is a substantial component of the antigenic epitope. Enzyme immunoassays which use synthetic cyclic citrullinated peptide (CCP) as the target antigen, offer a useful alternative to indirect immunofluorescence methods. Antibodies against CCP are predominantly of class IgG and are of diagnostic importance as a new specific marker especially during the initial stages of RA. In this review, the value of anti-CCP for the serodiagnosis of RA has been discussed. PMID- 14748271 TI - Dr. Nancy Morrison and her dying patient: a case of medical necessity. PMID- 14748272 TI - Reconciling private benefit and public risk in biotechnology: xenotransplantation as a case study in consent. PMID- 14748273 TI - A place for criminal law in the regulation of reproductive technologies. PMID- 14748275 TI - History of a gene patent: tracing the development and application of commercial BRCA testing. PMID- 14748274 TI - Industry and the academy: conflicts of interest in contemporary health research. PMID- 14748276 TI - Type 2 diabetes and children in Aboriginal communities: the array of factors that shape health and access to health care. AB - The human costs of unrestrained development on our traditional territory, whether in the form of massive hydroelectric development or irresponsible forestry operations, are no surprise for us. Diabetes has followed the destruction of our traditional way of life and the imposition of a welfare economy. Now we see that one in seven pregnant Cree women is sick with this disease, and our children are being born high risk or actually sick. PMID- 14748278 TI - [Intratympanic gentamicin treatment for intractable Meniere's disease]. AB - BACKGROUND: Intratympanic gentamicin is an accepted mode of treatment for patients with Meniere's disease who suffer from recurrent vertigo attacks in spite of conservative medical treatment. Recently, it gained popularity as the primary treatment for intractable Meniere's disease, prior to surgical procedures. However, administration methods and treatment protocols still differ from one medical center to another. OBJECTIVES: We aimed to present outcomes of gentamicin treatment for vertigo attacks and functional capacity, based on our experience with 23 patients over 3 years and review the literature on the subject. METHODS: Three specially prepared gentamicin injections, were administered once a month, according to the state of vertigo attacks, functional capacity, and hearing status. Clinical observation, frequent hearing examinations and caloric tests were used for follow-up. RESULTS: Following treatment, 20 (87%) patients no longer experienced vertigo attacks, while 16 of them (69.6%) reported full functional recovery. Two patients (8.7%) did not respond to treatment, and therefore, underwent surgical procedures. One patient (4.4%) did not complete therapy. Post treatment caloric tests showed significant deterioration (P < 0.02). Severe vestibular weakness (51-100%) was found in the treated ear in 11 of the 14 patients with unilateral disease (78.6%). The mean change in hearing examination was non-significant. However, two of the 21 patients suffered a change from moderate to severe hearing loss to complete deafness (9.5%). There was no case of post treatment infection or permanent tympanic perforation. CONCLUSIONS: Intratympanic gentamicin injection is a simple and safe procedure for intractable Meniere's patients, with a high success rate and low risk for hearing deterioration. It is recommended for patients with continuous functional disability prior to surgical treatment. PMID- 14748277 TI - [Lung and heart-lung transplantation in Rabin medical center: early experience with 70 cases]. AB - Lung transplantation is a relatively new field in solid organ transplantation. We present our early experience with the first 70 cases at the Rabin Medical Center during the years 1997-2003. Forty seven patients underwent single lung, eight double lung and eight heart-lung transplantations. The patients treated included 49 men and 21 women aged 5-66 years. There were 26 cases with emphysema COPD. 30 patients with pulmonary fibrosis. 5 patients with pulmonary hypertension/Eisenmenger and 9 patients with cystic fibrosis and bronchiectasis. Although early results (1997-1999) showed 1 and 3 year survival of only 50%, in the last 3 years (2000-2003), survival reached 84% and 82% at 1 and 3 years respectively. Improvement in the success rate is due to better patient selection, new immunosuppressive regimen and, most importantly, excellent teamwork. We conclude that lung transplantation is a viable option for selected patients with end-stage lung disease. PMID- 14748279 TI - [Characteristics of emergency room admissions of IDF soldiers in northern Israeli hospitals between May 2002 and April 2003]. AB - BACKGROUND: The hospitalization rate of patients visiting the emergency room depends on various parameters including demography, clinical data and other variables. Long term follow-up of hospitalized IDF soldiers reflected variability in hospital admission rate among hospitals and raises the possibility of vastly inappropriate hospitalizations. AIM: The aim of this study was to characterize the admission policy for IDF soldiers in various hospitals, to assess the rate of inappropriate hospitalizations, and to evaluate these costs to the Medical Corps. METHODS: Three hospitals in northern Israel were screened between May 2002 and April 2003, one urban tertiary care hospital (A) and two rural hospitals of different sizes (B and C). One fifth of the discharge forms were randomly screened. Every admission was given a risk evaluation based only on clinical data found upon presentation in the emergency room. One abnormal finding rendered the admission to be considered as appropriate. The rate of admission, lengths of stay, as well as clinical data and utilization of diagnostic tests were recorded and compared. RESULTS: During the study period 65 +/- 1105, 39 +/- 536, and 54 +/ 485 Emergency Room visits were made monthly in hospitals A, B and C, respectively. A total of 1.0% +/- 6.5. 1.4% +/- 12.6 and 2.7% +/- 11.8 of these visits resulted in hospitalization in hospitals A, B and C, respectively. Approximately one quarter of the patients admitted to hospital A were low risk, compared to more than half in the rural hospitals. Low risk admissions were one day shorter in the urban hospital, whereas high risk admissions had the same length of stay. The cost of low risk admissions in the urban hospital was about half of its cost in any of the rural hospitals. The estimated annual cost of low risk admission in all three hospitals screened was about $US 1 million. CONCLUSIONS: The admission rate in the urban hospital was half of its rate in the rural hospitals. Overall, 27% less low risk admissions were made in the urban hospital. The rural hospitals tendency to admit more low risk patients was compounded with their tendency to lengthen these admissions. The reason for this behaviour is not clear, although it may be related to differences in experience and clinical knowledge, consideration of the distance between the hospital and the soldier's home or unit, and financial admission policies. PMID- 14748280 TI - [Beer potomania]. AB - Beer potomania is a rare syndrome which is manifested by severe hyponatremia and various mental changes related to binge beer drinking and poor dietary intake. This article is a case study describing an alcoholic that was admitted because of this syndrome. We also discuss the pathophysiology of hyponatremia in alcoholics and especially in the syndrome of beer potomania. PMID- 14748281 TI - [Childhood-onset epileptic blindness--clinical correlates and outcomes]. AB - AIM: Acute blindness is a rare presentation of epileptic disorders referring to loss of sight without loss of consciousness corroborating with epileptic discharges recorded on the EEG. We summarized the pertinent literature on childhood-onset epileptic blindness. We also report on our overall experience with 26 children having developed epileptic amaurosis. This includes descriptions of the associated seizures. EEG abnormalities and reports on the response to anti epileptic therapy as regards to resolution of blindness and control of associated seizures. RESULTS: Our data for children with epileptic blindness is similar to previous reports regarding the reported duration of blindness and associated seizures, as well as the overall response to therapy and outcome. In our study, 25 children experienced acute episodes of complete visual obscuration lasting for 1-10 minutes and one 4-month-old infant had blindness from birth, representing status epilepticus amauroticus. Ten patients had accompanying generalized seizures, with a photosensitive response recorded in three cases. All of these children were treated with valproic acid regaining full vision and eight became seizure free. Ten children had accompanying focal motor seizures and unilateral temporo-posterior epileptic discharges recorded on EEG and two additional cases had isolated blindness and focal discharges. All 12 children were treated with carbamazepine, regaining full vision and complete seizure control in eleven. One infant with status epilepticus amauroticus since birth, secondary to a persistent epileptic focus over the right central-posterior areas, regained full vision following resection of an area of cortical dysplasia at the age of 8 months. Four additional children had the constellation of migraine headaches, focal motor seizures and complete blindness along with occipital EEG discharges, compatible with the syndrome of late-onset benign childhood epilepsy with occipital paroxysms (Gastaut syndrome). They were treated with carbamazepine and all became asymptomatic within a period of 1-4 years. CONCLUSIONS: Analysis of the literature, in addition to our overall experience, indicates that epileptic blindness in children is associated with a favorable outcome providing prompt diagnosis and treatment of the associated seizures or epileptic syndrome. This resulted in complete and long-standing resolution of blindness in all children together with satisfactory control of seizures. As such, we recommend a prompt EEG in any child presenting acute visual obscuration, even in the absence of additional overt epileptic phenomena. PMID- 14748282 TI - [Incidence of osteoporotic fractures in southern Israel]. AB - BACKGROUND: Very little is known about the epidemiology of osteoporotic fractures in Israel. AIM: We aimed to provide an estimate on the incidence of low-impact fractures in southern Israel. METHODS: The study included women and men 50 years and older with radiographic evidence of a new fracture. We screened and reviewed all the emergency room and hospital charts to identify all patients with low impact fractures who attended the Soroka Hospital during the corresponding months of January and February of 1998 through 2001. RESULTS: There were a total of 580 fracture patients (461 women and 119 men). The estimated incidence of all low impact fractures in the entire population aged 50 and older was 1064:100,000 (95% C.I. 981: 1153) per year. The estimated incidence of low-impact fractures in women was three fold higher than in men [1526:100,000 (95% C.I. 1390: 1675) vs. 490/100,000 (95% C.I. 408: 580) per year, respectively]. Fractures of the distal forearm and proximal hip (26% each) were, by far, the most common in women, followed by fractures of the humerus (18%), ankle (12%) and pelvis (5%). In men, proximal hip fractures (39%) were the most common, followed by fractures of the humerus (18%), distal forearm (15%) and ankle (14%). Multiple concurrent fractures occurred in 2.5% of the patients. CONCLUSIONS: Our data provides a preliminary estimate of the incidence of osteoporotic fractures, not including most vertebral collapse fractures, among women and men in southern Israel. Further studies are warranted to characterize fracture risk in other regions of the country and population sub-groups. PMID- 14748283 TI - [Evaluation of sperm samples from infertile men by flow cytometry]. AB - INTRODUCTION: Human spermatogenesis begins at adolescence and continues throughout life. This process includes morphologic, cytologic and biological changes, leading to the formation of mature spermatozoa. Male infertility may be caused by several reasons, including oligozoospermia at variable degrees and complete absence of mature spermatozoa. Routine spermatogram, measuring sperm counts, motility and morphology, might not provide complete information in the evaluation of these cases. This study is aimed to evaluate the possible use of flow cytometry in the identification of different sperm cell populations in sperm samples obtained from infertile men, and in determining the different cell types in various groups of infertile men. MATERIALS AND METHODS: Sperm samples from normal and infertile men (the latter were azoospermic or oligoteratozoospermic OTA) underwent flow cytometry analysis, after preparation with TNE buffer and staining with Propidium Iodide. The separation of germinal cells into different populations, according to their DNA content and chromatin condensation, was evaluated. The WINMDI (http://fac.-scripps.edu, J. Trotter) software was used for data analysis. RESULTS: Flow cytometric analysis enabled identification of several cell populations in sperm samples, including haploid, diploid and tetraploid cells. Certain cellular distribution patterns were observed in sperm samples from infertile men: mature haploid cells, diploid cells, domination of tetraploid or non-mature haploid cells, and combination of these patterns. These patterns appeared in a statistically different manner among fertile and infertile men; the median value of mature haploid cells was higher in normal men (91%, compared to 85% in the OTA group and 0% in the azoospermic men), while the median value of diploid and tetraploid cells was higher in azoospermic men (72% and 8.5% respectively, compared to only 1% and 0% in normal men). CONCLUSIONS: These findings suggest that flow cytometry of sperm samples may serve as a non-invasive tool for investigations of male infertility and for identification of appropriate candidates for interventional treatment. PMID- 14748284 TI - [Commercial sex engagement: medical and social aspects]. AB - Commercial sex engagement represents a common phenomenon that has not been studied as of yet. Relatively few surveys have been published worldwide. The image of the woman that takes part in commercial sex, as shown by these surveys, is similar to a female victim who was sexually abused during her childhood and who was exposed to maltreatment. It is also shown that these women frequently suffer from violence and that a high percentage of these females can be diagnosed as post-traumatics. In the literature, there is mention of their involvement in the spreading of veneral diseases. The state of affairs in Israel can be seen in the Parliamentary Investigation Committee for the Trading of Women and also in the article by BenZion et al. published in this issue of Harefuah. This paper shows the preliminary results of a study conducted among 55 women who are engaged in organized prostitution and reside illegally in Israel. Our editorial attempts to analyze the issue of commercial sex and the results of this research, and stresses certain important social and medical aspects of this subject. PMID- 14748285 TI - [Simple method to detect factors influencing drug expenditures]. AB - BACKGROUND: Analysis of drug expenditures as part of an assessment of various forms of management interventions is rarely reported. Several strategies have been used to decrease injudicious drug use: restricted availability of specific drugs under formulary instructions, educational programs for prescribing practices and streamlining to discontinue or limit therapy. Each intervention has limited effectiveness. OBJECTIVE: To detect factors influencing drug expenditures by a simple and accessible method. METHOD: Data analysis on drug utilization and expenditures was performed according to the "Pareto" technique and changes were calculated. Changes in expenditure due to inflation were also calculated; the cost value of each drug in 2000 was multiplied by its quantity used in years 2000 and 2001. Comparison of expenditures using this method of calculation enabled us to find the inflation rate of the drugs used in the hospital. RESULTS: Five important factors were found to have an impact on the pharmacy budget and calculations: drug inflation rate (-1%), new drugs introduced through the Pharmaceuticals & Therapeutic Committee (1.9%), extended usage of drugs due to new approved clinical indications (0.7%), opening of new services or widening of special old services (3.4%) and usage of commonly high volume prescribed drugs in various wards of the hospital (-8.9%). Quantities and costs for the 80 most expensive pharmaceuticals showed factors responsible for the drug expenditure when compared with the leading drug classes, as well as with leading drugs. Antimicrobials (27.5%), high volume IV infusions (11.0%), small volume infusions (8.5%), and anticoagulants (6.7%) were the most prominent drug classes that contributed to the high cost of the drug budget. Small volume infusions of Sodium Chloride 0.9% (7.8%), Piperacillin and tazobactam injections (5.6%). Ofloxacin & Ciprofloxacin drugs (4.7%), and Enoxaparin injections (4.6%) were the highest costing drugs. CONCLUSION: This simple method based on comparison of costs, utilization and judicious drug use, can provide accurate information that can be useful in planning and implementing a reasonable drug budget. PMID- 14748286 TI - [Commercial sex work: psychiatric and medical aspects]. AB - Commercial sex work has many medical and psychiatric implications. In recent years many papers on this matter have been published. However, many of these articles are based on opinions rather then on empiric fieldwork. Sex workers in the western world are not a major source of spread of venereal diseases. Nonetheless, it is important for medical decision makers to be aware of the health risks related to commercial sex work habits: for example. avoidance of condoms for oral sex, might be a risk factor for gonorrhea transmission. In field studies in Israel and abroad, it has been shown that sex workers are not always drug addicts or mentally ill; many of them see it as a legitimate form of work that is dictated by their needs, their social, personal and familial background. Commercial sex workers frequently suffer from emotional and somatic health problems that are often neglected. This review intends to examine the few empirical works that have been conducted on commercial sex workers. including our recent study performed in Israel. PMID- 14748287 TI - [Advanced cardiac life support--new answers to old dilemmas]. AB - Sudden cardiac death is a major health hazard in the western world. During the year 2000 new guidelines for advanced cardiac life support were published. These comprehensive guidelines are based on international consensus and present the scientific evidence for each therapeutic intervention. During the following two years new studies investigating the treatment of cardiac arrest have provided new evidence for patients' care. The innovations in this area include the use of automatic external defibrillators, new drugs and measures to minimize brain injury. This review covers some of the new emerging issues in the treatment of cardiac arrest. PMID- 14748288 TI - [Hypnosis: clinical applications]. AB - BACKGROUND: Hypnosis has a wide variety of applications in medicine. In recent years, recognition has grown for the need to subject the effectiveness of hypnosis to accepted research standards of clinical medicine. PURPOSE: This article aims to review salient studies of the effectiveness of hypnosis in several areas of medicine where hypnosis is commonly employed: analgesia, preparation for medical and surgical procedures, asthma, digestive disturbances, dermatological disorders, bleeding disorders, postsurgical and post chemotherapeutic nausea and vomiting, and smoking cessation. METHODS: This survey emphasizes studies that fulfill accepted standards of clinical research, including randomization, control groups, and adequate sample size. RESULTS: Firm empirical evidence is to be found for the effectiveness of hypnosis in analgesia. In the other areas reviewed there is also some evidence for the possible clinical value for hypnosis. CONCLUSIONS: Additional research will be necessary to determine the proper place of hypnosis in clinical medical care. PMID- 14748289 TI - [Therapeutic implications of endothelin antagonists for cardiovascular diseases]. AB - The endothelin system plays an important role in the pathophysiology of a variety of cardiovascular diseases including: congestive heart failure, essential and pulmonary hypertension, renal failure, and cerebrovascular disease. The biological effects of endothelin-1 on its target organs are mediated by two receptors: ETA and ETB. It is widely accepted that the vascular, cardiac, and renal adverse effects of ET-1 are mediated by ETA, while activation of ETB receptors leads to beneficial effects such as: attenuating the vascular and cardiac hypertrophic effects of ET-1 as well as the vasodilatory action of this peptide. In the last decade, a whole range of peptide and non-peptide ET-1 antagonists has been developed, some selective to ETA and others nonselective with dual antagonistic activity against both ETA and ETB. Several clinical studies have revealed that ET-1 antagonists are clinically beneficial therapeutic agents for the treatment of several cardiovascular diseases, leading to the approval of bosentan (ETA/ETB antagonist) for the treatment of pulmonary hypertension. The current review will focus on the recent developments in the endothelin field, with special emphasis on the ET-1 antagonist and their clinical use. PMID- 14748290 TI - [Prevention and pain management in term and preterm infants]. AB - Pain in the neonatal period is frequently experienced by 6-10% of newly born infants, preterm and term, who require neonatal intensive care. Repetitive painful procedures without adequate analgesia provided by the medical staff may bear long-term or even life-long adverse consequences. The use of pharmacological and non-pharmacological modalities in the management of pain may change this undesirable situation. The use of opioid analgesia for very painful procedures and the use of non-opioid medications in combination with opioids are essential. A change in the sensory environment of the sick infant is an important additional analgesic effect. In addition to pain management in the neonatal intensive care units, neonatal circumcision is the most frequent surgical procedure performed in males, and is frequently conducted without appropriate analgesia. The simple available methods of analgesia for neonatal circumcision are discussed and should be employed in order to avoid painful circumcision. Many pediatric medical associations in the developed world consider failure to provide proper routine analgesia for neonatal circumcision to be an unethical and sub-optimal medical practice. PMID- 14748291 TI - [Premature infant's nutrition--feeding strategies]. AB - Nutrition for preterm babies is aimed at achieving expected intrauterine growth and accretion of nutrients. Early aggressive nutrition in low birth-weight premature infants will prevent malnutrition and catabolic state. Early trophic feedings should be started as soon as possible for gastrointestinal priming. Enteral feedings are gradually increased as intravenous parenteral nutrition is weaned. Mother's (breast) milk is the best food for preterm babies, however it should be nutritionally enriched by powdered human milk fortifier. Preterm babies may also be fed with humanized cow milk based formulas, adjusted for the special needs of prematurity. Sucking skills usually mature around 34 weeks, corrected gestational age. Thus, small preemies are initially fed by orogastric tubes. Continuous gastric drip has no nutritional advantage, and most preemies tolerate bolus feedings well. The recommended daily increase in enteral feedings is up to 20 cc/kg/day, in order to achieve full enteral feedings fast enough, while trying to minimize the risk of necrotizing enterocolitis. As preterm babies mature, nutritive sucking is gradually introduced. Nutritive sucking is a complex action requiring coordination of sucking, swallowing and breathing. Nutritional assessment of preterm babies and nutritional recommendations upon discharge from the hospital are discussed. PMID- 14748292 TI - [Cystic fibrosis from the exocrine pancreatic point of view]. AB - Cystic fibrosis (CF) is the most common life-limiting autosomal recessive genetic disorder in Caucasians and is characterized by a wide variability of clinical expression. The vast majority of patients with CF have pancreatic insufficiency (PI) requiring exogenous pancreatic enzyme replacement therapy with meals. It is caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene, which encodes a protein that functions as a chloride channel and is regulated by cAMP. CFTR gene mutations may be classified as severe or mild with respect to pancreatic function status. Patients homozygous for two severe mutations experience severe clinical presentation including PI. Patients carrying at least one mild mutation are considered to be pancreatic sufficient (PS) and carry an overall prognosis that is vastly superior to CF patients with PI. Thus mild mutations appear to be dominant, providing enough functional CFTR to avoid PI. Five general mechanisms have been proposed which describe how CFTR gene mutations influence CFTR-mediated chloride secretion. Classes 1, 2, and 3. that confer little or no chloride channel function, confer the PI phenotype. In contrast, classes 4 and 5 allow CFTR residual function, that are expected to confer the less severe PS phenotype. Recent advances in mutation detection technology and the demonstration of characteristic abnormalities in trans epithelial potential difference measurements, have expanded the spectrum of diseases associated with the CFTR mutant genes, and paradoxically, in some ways complicated rather than simplified CF diagnosis. Until new diagnostic criteria is conclusively determined, CF diagnosis should be made on clinical rather than laboratory grounds. PMID- 14748294 TI - [Educating for quality healthcare--a conference held in Brisbane, Australia, July 2003]. PMID- 14748293 TI - [Depression, myocardial infarction and the immune system--the chicken before the egg problem]. AB - Major depression is a risk factor, associated with a twofold increase in the incidence of ischemic heart disease (IHD). One of every 6 patients suffers from major depression following acute myocardial infarction (AMI). This connection is of major concern, considering that major depression is an independent risk factor for cardiac morbidity and mortality after AMI, increasing overall mortality fourfold. Activation of the immune system has a significant role in the pathogenesis of IHD and depression. Vast physiological responses, mediated mostly by activation of the immune system, accompany post MI depression and may account for increased prevalence of arrhythmias and high mortality. This includes activation of the hypothalamic-pituitary-adrenocortical axis, endothelial dysfunction, platelets activation and alterations of phospholipid composition in cell membranes. On the other hand, activation of the immune system after AMI includes elevated levels of interleukin-1 and interleukin-6. which induce "sickness behavior", characterized by symptoms similar to those observed in major depression. The key question raised by this data, whether inflammation is the common ground for both AMI and depression, or if it is accompanying one and sets the ground for the other, remains unanswered at this time. The significance of major depression as an independent risk factor for post MI mortality and morbidity raises the practical question, whether treatment of depression can reduce mortality after AMI. Several recent studies that evaluated this presumption, failed to prove it. In this review we present an overview of the cross interaction between depression, AMI and inflammation and its diagnostic and therapeutic implications. PMID- 14748295 TI - Winners and losers under the new and improved Medicare. PMID- 14748296 TI - 10 new year's tips for finance leaders. PMID- 14748297 TI - Changing views about "usual charges". AB - The OIG's new definition of "usual charges" could have a significant impact on payment rates for a hospital's services. Traditionally, "usual charges" has referred simply to non-discounted rates maintained in a hospital's charge description master. The OIG's definition of the term includes discounted rates that a hospital negotiates with commercial payers. If the OIG's definition is adopted, payers may attempt to apply it, thereby complicating rate setting negotiations and monitoring of contract compliance. PMID- 14748299 TI - How are hospitals financing the future? Capital spending in health care today. PMID- 14748298 TI - Some gift-giving guidance. AB - Do Medicare beneficiaries need protection from providers bearing gifts? Yes, say Congress and the OIG through guidance on inducements to beneficiaries. PMID- 14748300 TI - The value of employee retention. PMID- 14748301 TI - Medicare gets much-needed shot in the arm. AB - The recently passed Medicare bill includes many provisions that will help to improve the nation's healthcare system: Allowing the private sector to deliver the prescription drug benefit by competing for consumers will result in greater choice at lower cost. Offering the option of health savings accounts promises to bring affordable coverage to the uninsured and will encourage healthcare consumers to make cost-conscious healthcare purchasing decisions. Giving hospitals an incentive to report outcomes will give consumers, for the first time, access to data they can use to make informed choices about healthcare providers. PMID- 14748302 TI - Five simple tools for managing labor costs. AB - Five tools can help healthcare organizations predict volume-related staffing variances and reduce labor costs: Staffing standards Position control Staffing calculator Real-time feedback loop Reporting. PMID- 14748303 TI - No more chances for supply-chain savings? Look again! AB - To optimize supply-chain performance, hospitals must: Take into account all elements of the supply chain; Decide who will lead the supply-chain improvement effort; Decide where to focus improvement initiatives; Assess the savings potential; Overcome barriers to success; Set appropriate targets and measure performance. PMID- 14748305 TI - The mystery of IT strategy. PMID- 14748304 TI - Physician specialty practices. Strategic survival for rural hospitals. AB - Rural hospital financial performance can be boosted by implementing a model for providing greater access to specialty care. Each of these models has its own unique financial implications and advantages for rural hospitals. PMID- 14748306 TI - What's a new patient worth to you? PMID- 14748307 TI - E/M code analysis can promote optimum ED payment. PMID- 14748308 TI - Credit balances. Spotlighting a little-known area of risk and opportunity. PMID- 14748309 TI - Synthesis and dissemination of research findings: a critical next step in the evolution of health promotion. PMID- 14748310 TI - A preliminary study on the impact of a pedometer-based intervention on daily steps. AB - PURPOSE: To investigate the effects of an 8-week, pedometer-based lifestyle physical activity intervention on physical activity levels. METHODS: Participants were 37 college employees who volunteered to participate in the study. The intervention consisted of goal setting, pedometer use, self-monitoring, and weekly e-mail reminders. Physical activity measures (pedometer, survey) were taken at baseline and immediately following the intervention. A t-test for paired samples was used to determine significant changes in measures. RESULTS: Results indicate a significant increase in average daily steps (p < .01), from 8565 (+/- 3121) steps at baseline to 10,538 (+/- 3681) steps after the program. Participants who averaged fewer than 6000 steps and obese participants experienced the greatest increase. DISCUSSION: Although preliminary, this study indicates that the minimal contact, self-managed, pedometer-based lifestyle intervention was effective in increasing the daily physical activity of participants. Given the combination of behavioral techniques used, a recommendation for future studies would be to examine the impact of individual behavioral techniques on daily physical activity. PMID- 14748311 TI - The effectiveness of nutrition education and labeling in Dutch supermarkets. AB - PURPOSE: Nutrition education and labeling may help consumers to eat less fat. The purpose of this study is to assess the effect of nutrition education with and without shelf labeling on reduced fat intake in Dutch supermarkets. METHODS: The design consisted of a randomized, pretest-posttest, experimental control group design. In total, 2203 clients of 13 supermarkets were included in the sample. Total fat intake of clients and behavioral determinants of eating less fat were measured by a questionnaire. A mixed-effect regression model was used for the analysis. RESULTS: No significant effects were found for the educational intervention, alone or with the labeling, on total fat intake and the psychosocial determinants of eating less fat. CONCLUSION: Nutrition education and labeling of low-fat food products in supermarkets did not prove to be effective strategies. The fact that the supermarket is a highly competitive environment may have accounted for this lack of effect. PMID- 14748312 TI - Tobacco use prevention and cessation policies in manufacturing facilities in the tobacco-growing state of Kentucky. AB - PURPOSE: To assess workplace tobacco use prevention and cessation policies in manufacturing facilities and explore factors associated with tobacco policies and practices in the tobacco-growing state of Kentucky. DESIGN: Cross-sectional, descriptive, correlational design. SETTING: Telephone survey of Kentucky manufacturing facilities. SUBJECTS: A total of 437 human resource managers (77% participation rate). MEASURES: Telephone interviews by trained local health department staff to assess indoor and outdoor smoking policies, sale of cigarettes on company property, and provision of cessation and prevention programs. RESULTS: Nearly seven in 10 manufacturing facilities had a written smoking policy, but only 43% banned indoor smoking. About one-fourth of companies reimbursed for cessation treatment and/or provided cessation resources. Companies with unions were more likely than those without unions to provide cessation resources but were also more likely to allow indoor smoking. Although large companies had more than two and a half times the odds as small companies to have a written smoking policy, they were more likely to allow cigarette sales on company property. CONCLUSION: Despite the importance of smoke-free policies in the workplace, most manufacturing facilities surveyed allowed indoor smoking and few helped smokers quit. Companies with unions were more likely to cater to their smoking employees. Manufacturing facilities provide an opportunity to protect large numbers of adult workers from the hazards of secondhand smoke and to provide quit assistance for smokers. PMID- 14748313 TI - Estimating the economic impact of secondhand smoke on the health of a community. AB - PURPOSE: This study provides a model to estimate the health-related costs of secondhand smoke exposure at a community level. MODEL DEVELOPMENT: Costs of secondhand smoke-related mortality and morbidity were estimated using national attributable risk values for diseases that are causally related to secondhand smoke exposure for adults and children. Estimated costs included ambulatory care costs, hospital inpatient costs, and loss of life costs based on vital statistics, hospital discharge data, and census data. APPLICATION OF THE MODEL: The model was used to estimate health-related costs estimates of secondhand smoke exposure for Marion County, Indiana. Attributable risk values were applied to the number of deaths and hospital discharges to determine the number of individuals impacted by secondhand smoke exposure. RESULTS: The overall cost of health care and premature loss of life attributed to secondhand smoke for the study county was estimated to be $53.9 million in 2000-$10.5 million in health care costs and $20.3 million in loss of life for children compared with $6.2 million in health care costs and $16.9 million in loss of life for adults. This amounted to $62.68 per capita. CONCLUSIONS: This method may be replicated in other counties to provide data needed to educate the public and community leaders about the health effects and costs of secondhand smoke exposure. PMID- 14748314 TI - Associations of location and perceived environmental attributes with walking in neighborhoods. AB - PURPOSE: Ecological models highlight the importance of environmental influences. We examined associations of coastal versus noncoastal location and perceived environmental attributes with neighborhood walking, total walking, and total activity. METHODS: Telephone interviews with 800 faculty and general staff of an Australian university. RESULTS: Men were significantly more likely to walk in their neighborhood if they lived in a coastal location (odds ratio [OR] = 1.66), and they highly rated environmental "aesthetics" (OR = 1.91), "convenience" of facilities (OR = 2.20), and "access" to facilities (OR = 1.98). For women, neighborhood walking was associated with high ratings of "convenience" (OR = 3.78) but was significantly less likely if they had high ratings for "access" (OR = 0.48). For total walking and total physical activity, few significant associations emerged. CONCLUSIONS: Environmental attributes were related to walking in the neighborhood but not to more general activity indices. Understanding gender-specific environmental correlates of physical activity should be a priority. PMID- 14748315 TI - Availability of school physical activity facilities to the public in four U.S. communities. AB - PURPOSE: This study documents the public availability of school physical activity facilities, reasons facilities were not made available to the public, and the barriers and benefits associated with having facilities available. DESIGN: Cross sectional. SETTING: All schools located in four geographic locations (Washington County, Maryland; northwest suburbs of Minneapolis, Minnesota; Jackson, Mississippi; and Forsyth County, North Carolina). SUBJECTS: The survey was completed by 289 school representatives for a response rate of 82.3% (289/351 schools). For the nonrespondents, 61 of 62 schools were visited to obtain information on outdoor facilities. MEASURES: Information on indoor and outdoor physical activity facilities available to the public was collected for all schools (public, private, and colleges) using a telephone survey. Site visits were made to schools that did not participate in the survey to collect information on outdoor facilities only. RESULTS: The schools in the four geographic areas owned a wide range and number of physical activity facilities for their students. For the schools, 27% had no indoor facilities and 11% had no outdoor facilities. Private schools and colleges were less likely to have indoor or outdoor facilities compared to public schools. Outdoor facilities were available more often to the public than indoor facilities overall, across sites, and by school type (public, private, college). Among the 313 schools with outdoor facilities, 240 (77%) allowed at least some public use, and among the 210 schools with indoor facilities, 134 (64%) allowed at least some public use. Some reasons that facilities were not made available included: for student use only, supervision and personnel requirements, safety concerns, insurance, liability, and a private or church-owned status. The most common benefits of allowing the public to use the facilities included providing a space to keep youth active and good publicity for the school. CONCLUSION: These findings suggest that safety, insurance, and liability concerns are barriers that need to be addressed with schools before indoor and outdoor facilities can be made available to the public. Furthermore, emphasizing the benefits that we found, such as providing a space to keep youth active and good publicity for the school, would also be important. PMID- 14748316 TI - Association of environmental factors to meeting physical activity recommendations in two South Carolina counties. AB - PURPOSE: The purpose of the study was to examine associations between environment and policy factors and physical activity. DESIGN: A random-digit-dialed, cross sectional telephone survey was administered. SETTING: The setting was a two county area of eastern South Carolina. SUBJECTS: Before weighting, the sample included 1936 adults; 36.9% African-American, 63.1% white, and 60.1% women. The age group distribution was 28.8% 55+ years, 39.3% 35-54 years, and 31.9% 18-34 years of age. The response rate was 62.9%. MEASURES: Six physical activity questions (2001 Behavioral Risk Factor Surveillance Survey [BRFSS]) were used to create a dischotomous variable, "meets/does not meet recommendation for moderate or vigorous physical activity." Self-report items assessed knowledge, presence, and use of recreational facilities; presence of environmental and worksite supports; perceived safety; condition of sidewalks; and quality of street lighting. RESULTS: Linear and logistic regression were used to analyze the data. Unadjusted odds for meeting the recommendation were significantly greater for well-maintained sidewalks (OR = 1.90); safe areas for walking/jogging (OR = 1.39); knowledge of routes for bicycling (OR = 1.38) and walking/jogging (OR = 1.32); and worksites with sports teams (OR = 1.53), exercise facilities (OR = 1.33), flexible time for exercise (OR = 1.33), and preventive checkups (OR = 1.26). Among persons who met the recommendation, means were greater for number of known walking/jogging routes (p = .04); number of known bicycling routes (p < .01); number of days per month uses tracks, trails, routes, pathways (p < .01); and number of days per month uses outdoor recreation areas (p < .01). CONCLUSION: The results support an association between level of physical activity and environmental and policy factors in two southeastern counties in South Carolina. Limitations of the study include self-reported data and cross-sectional design. PMID- 14748317 TI - Zoning should promote public health. AB - Legally, governments use their police powers to protect public health, safety, and welfare through zoning. This paper presents a case for revisiting zoning on the basis of increasing evidence that certain types of community design promote public health, as opposed to the dominant pattern of sprawl development, which does not. Zoning, and the land use planning linked to it, that prohibits or disfavors health-promoting community designs contradicts the inherent public policy goal on which it is based. If there is a paradigm shift underway, from traditional sprawl to health-promoting community designs, then health professionals and others should understand why zoning must be reassessed. PMID- 14748318 TI - Understanding the relationship between health risks and health-related costs. AB - Dr. Anderson reflects on the lack of consistency in studies on the relationship between health risks and medical costs. Part of the inconsistency is caused by differences in risk assessment instruments, but a more important concern is lack of clear conceptualization and statement of research goals. One reason for exploring this relationship is to be able to predict health care costs accurately: a very different reason is to identify causal variables, the modification of which will have an impact on costs. PMID- 14748319 TI - Association of additional health risks on medical charges and prevalence of diabetes within body mass index categories. AB - PURPOSE: The prevalence of obesity and diabetes continues to increase among employee populations. Although medical costs and the prevalence of diabetes have been studied across increasing body mass index (BMI) categories, little attention has been given to the association of additional health risks within those categories. The purpose of this study was to examine the association of health risk levels on medical charges and prevalence of diabetes across BMI categories within an employee population. METHODS: A cross-sectional study design utilized health risk appraisal data (30% response rate) to measure BMI levels, self reported diabetes status, and selected additional health risks among 38,841 active employees under age 65 of the General Motors Corporation. Associated average annual medical charges from 1996 to 2000 were calculated for defined health risk levels across five BMI categories (< 18.5; 18.5-24.9, 25-29.9, 30 34.9, and > 35). RESULTS: Higher medical charges were significantly associated with additional health risks (zero risks to four or more risks) across each of the BMI categories: $2689 to $7576 (< 18.5); $2655 to $6555 (18.5-24.9); $3239 to $7118 (25-29.9); $3579 to $7758 (30-34.9); and $4151 to $8075 (> or = 35). Likewise, higher prevalence of diabetes was significantly associated with additional health risks (zero risks to four or more risks) across the BMI categories: 2.6% to 7.0% (< 18.5); 1.3% to 2.7% (18.5-24.9); 2.4% to 5.3% (25 29.9); 5.5% to 8.3% (30-34.9); and 7.7% to 15.8% (> or = 35). DISCUSSION: Medical costs and the prevalence of diabetes were lower when the numbers of additional health risks were lower, regardless of the BMI category. Programs to promote weight management have largely been unsuccessful in maintaining long-term weight control. The current results suggest that a strategy focused on reducing health risks within any weight category could provide an alternative strategy to achieve medical cost savings and a lower prevalence of diabetes. The implied benefits of risk reduction within BMI categories would need to be confirmed with a longitudinal study. PMID- 14748320 TI - [The use and abuse of ICD-10--a critical view from a gastroenterologist]. PMID- 14748321 TI - [Multipotent adult progenitor cells and their application to regenerative medicine]. PMID- 14748322 TI - [Manipulating the differentiation and regeneration of the gastrointestinal epithelia--prospects for clinical application]. PMID- 14748323 TI - [A case of acute phlegmonous gastritis followed up by endoscopic ultrasonographic examination]. PMID- 14748324 TI - [Anti-CD20 antibody and combination chemotherapy followed by autologous peripheral blood stem cell transplantation after high-dose chemotherapy induced complete remission in a case of multiple lymphomatous polyposis of the gastrointestinal tract]. PMID- 14748325 TI - [A case of liver abscess with septic thrombophlebitis of the portal vein and superior mesenteric vein caused by Streptococcus milleri]. PMID- 14748327 TI - [Two cases of asymptomatic primary biliary cirrhosis with ascites]. PMID- 14748326 TI - [An autopsy case of the hepatocellular carcinoma associated with multiple myeloma which developed fatal massive hemolysis due to the Clostridium perfringens septicemia following TAE]. PMID- 14748328 TI - [A case of hepatocellular carcinoma available to transcatheter arterial treatment by divided median arcurate ligament compressed celiac artery]. PMID- 14748329 TI - Nigeria: HIV-positive woman launches suit after being denied treatment. AB - A 39-year-old woman who was denied treatment at a hospital in Lagos simply because she is HIV-positive is fighting back through the courts. The Center for the Right to Health has filed a lawsuit on her behalf, seeking damages as well as an injection to prevent further discrimination against people with HIV/AIDS who seek treatment. PMID- 14748330 TI - Court sets aside immigration employment authorization containing HIV-related job restrictions. AB - In an order issued on 30 May 2003, the Federal Court-Trial Division set aside an employment authorization that prohibited an HIV-positive man from working in childcare, primary and secondary school teaching, health services, and agriculture. The court directed immigration medical officers to decide whether a new medical examination is required. This is a significant case for HIV-positive immigrants and refugees who are required to hold an employment authorization if they intend to work in Canada while their applications for permanent residence are decided, a process that can take years. PMID- 14748331 TI - Immigration removal order stayed on humanitarian and compassionate grounds. AB - The Immigration Appeal Division of the Immigration and Refugee Board (IRB) has stayed a removal order against a man living with HIV and HCV on humanitarian and compassionate grounds. Taking an enlightened approach, the IRB found that Gurbinder Randhawa's efforts at rehabilitation for his drug dependence was a positive factor in staying the removal order. PMID- 14748332 TI - Immigration Act medical inadmissibility provision survives constitutional challenge. AB - In a decision issued on 27 June 2002, the Federal Court of Canada ruled that the medical inadmissibility provision of the Immigration Act did not infringe sections 7 or 15 of the Canadian Charter of Rights and Freedoms. Although the Immigration Act was repealed effective 28 June 2002 with the coming into force of the Immigration and Refugee Protection Act (IRPA), the court's reasons are relevant to some HIV-positive people seeking admission to Canada under the new legislation. PMID- 14748333 TI - Fear of lack of adequate medical care not well-founded fear of persecution. AB - The Federal Court-Trial Division refused to set aside the Immigration and Refugee Board's (IRB) decision that a refugee claimant was not a Convention refugee because she had not proven a well-founded fear of persecution based on her HIV status or AIDS diagnosis. The court emphasized that the fear of a lack of adequate medical treatment, without a clear link to a ground under the United Nations Convention relating to the Status of Refugees, does not constitute a well founded fear of persecution. PMID- 14748334 TI - New tainted-blood class actions in Quebec and Alberta. AB - In two cases in Quebec and Alberta, people infected with HIV and HCV through infected blood and blood products have successfully defended motions to strike out all or parts of their legal actions against federal and provincial governments and the Canadian Red Cross Society (CRCS). On 16 January 2003, the Quebec Superior Court ruled that the plaintiffs in a class action could rely on the Krever Commission Report in their application for certification of a class proceeding. On 20 February 2003, the Alberta Court of Queen's Bench dismissed an application brought by the Canadian and Alberta governments to strike out the legal action brought against them. These cases illustrate that the settlement schemes proposed by the federal and provincial governments and the CRCS, and approved by the courts, have not put an end to the civil and constitutional claims brought by people seeking compensation for infection through tainted blood. PMID- 14748335 TI - Judge refuses to reduce damage award to HIV-positive man. PMID- 14748336 TI - Infections rates in penitentiary influence judge's decision not to transfer young offender to adult court. PMID- 14748337 TI - Australia: court orders doctors to pay damages to woman who contracted HIV from her husband. AB - IN a case that received extensive media coverage, the New South Wales Supreme Court has found two doctors negligent for failing to take steps to make a bride aware that she was at risk as a result of her prospective husband's HIV infection. The court awarded the woman AUD$727,437 in damages. The couple had attended the physicians together to be tested for HIV and other sexually transmitted infections, but had received their test results separately. She subsequently contracted HIV as a result fo unprotected sexual intercourse with him. PMID- 14748338 TI - European Court of Human Rights accepts blood donor screening based on sexual orientation. AB - In a series of three rulings released simultaneously on 15 October 2002, the European Court of Human Rights dismissed complaints that Italy's approach to screening blood donors infringed privacy rights and discriminated against lesbians and gay men. PMID- 14748339 TI - US: court rules deportation of HIV-positive Dominican does not violate convention against torture. AB - The US District Court for New York has upheld a tribunal decision deporting an HIV-positive Dominican national. The court held that the tribunal did not err in finding that Rogelio Reyes-Sanchez had failed to show that he would more likely than not be subject to torture if deported. The court agreed that evidence suggesting that people living with HIV/AIDS may receive substandard medical treatment from public and private hospitals in the Dominican Republic di not constitute evidence of torture. PMID- 14748340 TI - UK: Welsh court reduces sentence, cites HIV status. AB - A Welsh appeal court has reduced the sentence handed down to an offender because of his HIV status, despite his lengthy criminal record. The court reduced the sentence from five to three-and-a-half years' imprisonment. PMID- 14748341 TI - Asylum seeker with AIDS loses fight against relocation within the UK. AB - An English court has upheld the decision to relocate an asylum seeker living with AIDS from London to Leeds. The court decided that the public interest in housing asylum seekers where accommodation is available outweighed the woman's interest in remaining in London, where she had an established support network, including medical care. The government has undertaken not to implement the order until her application for asylum is ruled on. PMID- 14748342 TI - [Advances in the study of Momordica charantia L]. PMID- 14748343 TI - [Special features of clinical effects of an angiotensin converting enzyme inhibitor lisinopril in patients with arterial hypertension]. PMID- 14748344 TI - A profile of success and failure in telehealth--evidence and opinion from the Success and Failures in Telehealth conferences. AB - Papers presented at the three Successes and Failures in Telehealth conferences have made a helpful contribution to what is known about the practicalities of implementing and using telehealth. Ten of the 89 papers covered general concepts, 40 described primary studies and 26 discussed telehealth programmes or networks. Common themes that emerged on determinants of success and failure included the reliability of equipment, the reliability of vendors, political and budgetary issues,the perceived need for telehealth, the involvement of stakeholders, health professionals' opinions, the stability of management structures, the turnover of personnel and cooperation between organizations. Limitations in some papers included an absence of data, weak study designs, short-term perspectives and lack of information on health outcomes. Despite these limitations, the papers have made a valuable contribution to our knowledge of what has worked and what has failed in the field of telehealth. PMID- 14748345 TI - [Chronic heart failure and atrial fibrilation: aspects of management]. PMID- 14748346 TI - Consultation-Liaison Psychiatry Literature Database (2003 update). Part I: Consultation - Liaison Literature Database: 2003 update and national lists. AB - Every day there are 10,000 scientific articles published. Since the Consultation Liaison ("C-L") psychiatrist may be asked to consult on a patient with any medical illness, e.g., severe acute respiratory syndrome (SARS), malaria, cancer, stroke, amytrophic, lateral sclerosis, and a patient who may be on any medical drug, methods need to be developed to review the recent literature and have an awareness of key and essential current findings. At the same time, teachers need to develop a current listing of seminal papers for trainees and practitioners of this newest cross-over subspecialty of psychiatry-now called Psychosomatic Medicine. Experts selected because of their writings and acknowledged contributions to a specific clinical area or problem hope examined thousands of citations to choose those articles, chapters, books, or letters that they regard as most important to Psychosomatic Medicine. In addition, psychiatric specialists in six countries have provided their national Psychosomatic Medicine (Consultation-Liaison) lists as examples of what they regard as the most important teaching materials journals: Australia, Brazil, Greece, Mexico, Portugal, and Taiwan. It is our belief that a cogent, international, systematic review will provide the greatest success in creating a "regionally appropriate" teaching and consultation literature database with world-wide applicability. We review our current progress on this literature database and software, the technical system and data organization involved, the approach used to populate the literature system, and ongoing development plans to bring this system to the physician via mobile technologies. PMID- 14748347 TI - Consultation-Liaison Psychiatry Literature Database (2003 update). Part II: topics, contributors, and database. PMID- 14748348 TI - Gone but not forgotten. PMID- 14748349 TI - Management challenge. Test intentions. PMID- 14748350 TI - A novel mutation with Ins C (882-883) of the factor X gene in a Taiwanese Chinese factor X-deficient family. PMID- 14748352 TI - New editorial partnerships and international awards in integrative medicine. PMID- 14748354 TI - Early nonresponse for tenofovir regimen. PMID- 14748353 TI - Reconstruction of radioactive plume characteristics along Chernobyl's Western Trace. AB - Using data obtained from 435 radiation sampling stations in the Red Forest, 1.5 km W if the Chernobyl Nuclear Complex, we reconstructed the deposition pathway of the first plume released by the accident, Chernobyl's Western Trace. The dimensions and deposition rates of the plume remain sharply defined 15 years after the accident. Assuming a uniform particle distribution within the original cloud, we derived estimates of plume dimensions by applying geometric transformations to the coordinates at each sample point. Our derived estimates for the radioactive cloud accounted for 87% of the variation of radioactivity in this region. Results show a highly integrated bell-shaped cross-section of the cloud of radiation, approximately 660 m wide and 290 m high, traveling at a bearing of 264 degrees from reactor IV. Particle sizes within Chernobyl's Western Trace were within the most dangerous range for inhaled aerosols (2-5 microm). Therefore, reconstruction of the dispersion of such particles is critical for understanding the aftermath of nuclear and biological aerosol releases. PMID- 14748355 TI - Efficacy demonstrated for once-daily regimen. PMID- 14748356 TI - Benefits of HAART in co-infected patients. PMID- 14748357 TI - Italian study investigates value of STI. PMID- 14748358 TI - Age-related prevalence of HPV in men. PMID- 14748359 TI - Broccoli compound inhibits HSV. PMID- 14748360 TI - New oral drug for PCP. PMID- 14748361 TI - Growth hormone does not affect viral load. PMID- 14748362 TI - Those who marched. Lystra E. Gretter, RN (1858-1951). PMID- 14748363 TI - Package insert revisions for ritonavir. PMID- 14748364 TI - HIV risk greater for uncircumcised men. PMID- 14748365 TI - Lesbians and Pap smears. PMID- 14748366 TI - Catholic Church makes news about condoms. PMID- 14748367 TI - New HPV test now recommended. PMID- 14748368 TI - Website aimed at HIV-positive women. PMID- 14748369 TI - Marijuana compound effective against pain. PMID- 14748370 TI - Analysis. Space invaders. PMID- 14748371 TI - Train of thought. PMID- 14748372 TI - What works. On primary care. PMID- 14748374 TI - [A review of adverse effects of Panax notoginseng (Burk.) F.H.Chen]. PMID- 14748373 TI - [Advances in pharmacological study of Piper kadsura (Choisy) Ohwi]. PMID- 14748376 TI - Ageing and health. PMID- 14748375 TI - [Advances in the study of Paris L. and its uses]. PMID- 14748377 TI - Lipid profile of Indian children and adolescents. AB - Dyslipidaemia when diagnosed in childhood predicts the development of clinical atherosclerotic disease in adulthood. Ever since we became aware of the abnormally high incidence of coronary artery disease amongst Indians, there always has been an ever-growing need for study of lipid values amongst Indian children and adolescents. Five hundred and eighty-six children (352 boys and 234 girls) from a public school aged between 8 and 18 years (mean age: 12.86 for boys and 12.15 for girls) were included in the study for assessment of lipid profile during the period from April, 1999 to March, 2000. Fasting venous blood was drawn from each child and total cholesterol and triglyceride levels were measured by enzymatic method, high-density lipoportein (HDL)-cholesterol was measured using specific precipitation method and lipoprotein (a) [Lp (a)] was measured by immunoturbidimetric method. After biochemical estimation was completed reference values and percentiles were calculated for each lipid component. Mean cholesterol and triglyceride values were higher amongst girls compared to boys. HDL cholesterol values were lower amongst Indian boys compared to girls. Mean Lp (a) values were higher amongst both Indian boys and girls and more so amongst girls compared to boys. Since Lp(a) values remain unchanged through ages, these values should represent corresponding values in adult Indian males and females. It is being expected that data presented in this study should form guidelines with regard to dietary and pharmacological intervention for dyslipidaemia amongst children and adolescents. PMID- 14748378 TI - Arthritis in the elderly. AB - Joint disorders are common in the elderly population. Commonest conditions are rheumatoid arthritis, osteo-arthritis and crystal induced arthritis. Comorbid events in the elderly patients make disease management even more difficult. Some tests are needed to distinguish these conditions from more serious diseases like carcinomatous arthritis or multiple myeloma. Management strategies are usually similar to that of younger onset arthritis with dose adjustment of drugs due to underlying age related hepatic and renal dysfunctions. PMID- 14748379 TI - Cancer in the elderly. AB - Cancer, the dreaded killer disease, has its aetiology in several factors like genetic, environmental, diet habits, etc. Age over 65 years is generally considered elderly and the cancer incidence increases with age. Cancer is second to cardiovascular disease for the cause of death in the elderly. The behaviour of certain cancers also differ in the elderly thus adding to the ultimate outcome that is usually confounded by several comorbid illnesses. The prognostic factor varies with type of cancer. Older patients with Hodgkin's disease and acute myeloid leukaemia do worse than breast cancer. Radiation therapy is well tolerated by the elderly. PMID- 14748380 TI - Falls in the elderly--clinician's approach. AB - Fall is an involuntary event producing a change in posture resulting in the individual adopting an unplanned supine position. Globally more than one-third of persons 65 years of age or older fall each year and in half of such cases the falls are recurrent. Several predisposing factors for such falls have been recognised like age related changes in posture control, reduced visual acuity, anxiety, drugs, environmental hazards and underlying neurological diseases. It is the interplay of predisposing and precipitating factors that really matter and not exactly a single cause. Evidence based interventions have been suggested from recent clinical trials and certain preventive guidelines are present to reduce the rate of falling, provided a periodic targeted approach is followed. PMID- 14748381 TI - Endothelium, atherosclerosis and calcium channel blockers. AB - Probably the most significant advance in cardiovascular medicine over the last two decades has been the identification of endothelial cells as a vasoactive organ. The endothelium plays a primary autocrine/paracrine regulatory role by secreting substances that control both vascular tone and structure. The dysfunctioning endothelium, which is characteristic of essential hypertension and most of the cardiovascular risk factors, is a major promoter for atherothrombosis and, consequently, cardiovascular treatment. One of most relevant mechanisms of endothelial dysfunction is oxidative stress production, which causes nitric oxide breakdown. The clinical manifestations of atherosclerosis are by far the prevailing cause of morbidity and mortality in hypertensive patients. Various clinical studies have shown the beneficial effects of calcium channel blockers on endothelial dysfunction. The potential mechanism by which calcium channel antagonists could exert their beneficial activity on endothelial dysfunction is very unlikely to be a calcium-dependent mechanism since endothelial cells do not express voltage-operated calcium channels. Experimental evidence suggests that calcium channel antagonists exert an anti-oxidant effect and therefore could protect endothelial cells against free-radical injury. Nifedipine is the calcium channel blocker, which improves endothelial nitric oxide availability, antagonises endothelin 1, restores endothelial permeability and low-density lipoprotein deposition. Calcium antagonists have demonstrated anti-atherogenic properties in various dinical studies. Calcium antagonists--in addition to their primary action in lowering blood pressure influence numerous cellular process involved in early atherogenesis. All calcium channel blockers do not offer same benefit of reversal of endothelial dysfunction. Amlodipine does not seem to be as effective as nifedipine in terms of atheroprotection. With nifedipine, the overall risk of cardiovascular events decreased significantly. Therefore, nifedipine (long-acting formulation) remains the first choice calcium channel blocker in cardiovascular therapeutics--today and tomorrow. PMID- 14748382 TI - A multicentric study to evaluate efficacy and safety of cefetamet pivoxyl in lower respiratory tract infections in Indian patients. AB - This multicentric, open label, non-comparative study was designed to evaluate the extended spectrum of third generation oral cephalosporin, cefetamet pivoxyl in the treatment of patients with lower respiratory tract infections. This study was conducted among 111 patients with clinical, radiological and bacteriological findings consistent with the diagnosis. After obtaining written informed consent, patients were given cefetamet 500 mg tablet twice a day for 7 days. Cefetamet consistently decreased all clinical signs and symptoms at post-therapy visit. All the treated patients were either cured or improved. Cefetamet was well tolerated with a low incidence of drug related adverse events. The findings of this study indicate that cefetamet pivoxyl was well tolerated and is suitable option for the treatment of patients with lower respiratory tract infection. PMID- 14748383 TI - Efficacy and safety of the first parenteral selective COX-2 inhibitor, parecoxib sodium, in adult patients with postoperative pain. AB - Parecoxib, a prodrug of valdecoxib, a selective COX-2 inhibitor, has been recently introduced for the treatment of moderate to severe postoperative pain. This prospective, open, multicentric study enrolled 260 patients undergoing orthopaedic, gynaecological, dental and general surgery. Postoperatively, patients were treated with parecoxib, 40 mg IM/IV. There was a statistically significant decrease in the mean pain intensity score (p<0.05). At the end of 24 hours, 89.6% of total cases had a very good to total relief of pain. The mean duration of analgesia was 19.26 hours and mean time of onset of analgesia was 16.25 minutes ranging from 11-20 minutes. The laboratory values were within normal limits. The drug was well tolerated. There was no report of any hypersensitivity reaction. This study suggests that parecoxib, in a dose of 40 mg IM/IV, is an effective and safe option for the management of postoperative pain. PMID- 14748384 TI - MgATP-induced conformational changes in a single myosin molecule observed by atomic force microscopy: periodicity of substructures in myosin rods. AB - This paper discusses the conformational changes in a single myosin molecule directly observed using atomic force microscopy (AFM). The myosin molecules were pretreated in rigor solutions without MgATP or in relaxed solutions with various concentrations of MgATP. The images of these molecules were obtained using a tapping mode AFM. The results indicate that the orientation of the myosin's heads and tail strongly depend on the MgATP concentration. Without using MgATP, almost all of the myosin molecules are in the extended form; however, when MgATP is used, the molecules bend according to the level of MgATP concentration. The mean square end-to-end distance of the myosin molecules is significantly shorter with p[MgATP] = 4 than with p[MgATP] = 6. The rod region did not show the same level of intensity along their length in the extended form. The rods exhibited clusters of discontinuity, which were identified as substructures. The size of these substructures change at intervals that are multiples of 14.3-14.5 nm, which reflects the periodicity of the alpha-helical coiled coils. The substructure clusters also correspond to the myosin crossbridge spacing in muscles (14.3 or 43 nm). These results suggest that the myosin's head bends in conjunction with the bending or tilting in the helical substructures. Conformational changes of the myosin molecule induced by MgATP seem to mimic the molecular motions in a muscle's force generation process. PMID- 14748385 TI - Calibration and quantification of fast intracellular motion (FIM) in living cells using correlation analysis. AB - Video rate confocal laser scanning microscopy at the highest spatial and temporal resolution of backscattered light (BSL) imaging allowed for regular observation of fast intracellular motion (FIM) first revealed in living neoplastic cells. However, the absence of an objective evaluation has hampered further study of the mechanisms and biological significance of FIM. Particularly, a quantification of apparent differences in velocities that would complement and improve the current demonstration of FIM by color coding using the combination of red-green-blue (RGB) images had been missing. Standard methods of tracking or pattern recognition could not be applied because of the fuzzy nature of images of FIM. A search for a suitable method led to correlation analysis. It was calibrated on Brownian motion and a known type of motion, such as cell marginal ruffling, compared with FIM. Results approved its explanatory potential. Therefore, several crucial incidences of FIM could be analyzed. Apart from an argument against viewing FIM as a manifestation of simple Brownian motion, the correlation analysis of FIM in the adjacent peripheries of a rat fibroblast and a K4 rat sarcoma cell confirmed the notion of higher and uneven distribution of velocity of FIM in a tumor cell so far shown in color-coded images only. This result and other yet unpublished observations indicate that the velocity and topology of FIM can also contribute to a biological distinction between neoplastic and normal cells. Regular application of the correlation analysis should further expand the study of FIM for its mechanisms and predictive value. Such an approach should be thoroughly examined for a contribution to the knowledge of cancer cells. PMID- 14748386 TI - Possibility of charge contrast imaging of polymeric materials. AB - Preliminary results illustrate the possibility of charge contrast imaging (CCI) of polymeric materials. Possible CCI images of low-density polyethylene and polyvinyl chloride reveal details that may aid in the characterization of the microstructure of polymeric materials. These pictures were obtained with a Hitachi S-3000N variable pressure scanning electron microscope with the environmental secondary electron detector (ESED). PMID- 14748387 TI - A differentially pumped secondary electron detector for low-vacuum scanning electron microscopy. AB - A new design of secondary electron (SE) detector is described for use in low vacuum scanning electron microscopes. Its distinguishing feature is a separate detector chamber, which can be maintained at a pressure independent of the pressure in the specimen chamber. The two chambers are separated by a perforated membrane or mesh across which an electric field is applied, making it relatively transparent to low-energy electrons but considerably less so to the gas molecules. The benefits of this arrangement are discussed. The final means of detecting the electrons can be a conventional scintillator and photomultiplier arrangement or any of the methods using the ambient gas as an amplifying medium. Images obtained with the detector show good SE contrast and low backscattered electron contribution. PMID- 14748389 TI - Three-dimensional architecture of ribosomal DNA within barley nucleoli revealed with electron microscopy. AB - To elucidate the topological positioning of ribosomal RNA genes (rDNA) and nucleolar structure in three dimensions, we examined the localization of rDNA using in situ hybridization (ISH) analysis by scanning electron microscopy (SEM). The rDNA genes within the three-dimensional architecture of nucleoli were detected on chromatin fibers that connect a thick strand-like structure and a protrusion of rDNA into the inner nuclear hole where the nucleolus is formed. This novel use of ISH together with SEM is useful for the analysis of nucleolar structure in detail. Furthermore, rDNA was detected at the periphery of the fibrillar centers (FCs) of the nucleolus using immuno-gold labeling together with transmission electron microscopy (TEM). In situ hybridization with TEM confirmed that rDNA is naked and thus active in the FCs of nucleoli; ISH with SEM confirmed that rDNA is not covered with ribonucleo proteins at the protruding point and is thus inactive. We also show that the distribution pattern of FCs differs from sample to sample. These results indicate that rDNA is transcribed dynamically in a time- and region-specific manner over the course of the cell cycle. PMID- 14748388 TI - Confocal microscopic image sequence compression using vector quantization and three-dimensional pyramid. AB - The three-dimensional (3-D) pyramid compressor project at the University of Glasgow has developed a compressor for images obtained from confocal laser scanning microscopy (CLSM) device. The proposed method using a combination of image pyramid coder and vector quantization techniques has good performance at compressing confocal volume image data. An experiment was conducted on several kinds of CLSM data using the presented compressor compared with other well-known volume data compressors, such as MPEG-1. The results showed that the 3-D pyramid compressor gave a higher subjective and objective image quality of reconstructed images at the same compression ratio, and presented more acceptable results when applying image processing filters on reconstructed images. PMID- 14748390 TI - Software acceleration techniques for the simulation of scanning electron microscope images. AB - A scanning electron microscope (SEM) simulator was developed based on the models used in the MONSEL software. This simulator extends earlier work by introducing an object-oriented framework and adding optimization methods based on precomputation of electron trajectories. Several optimizations enable speedup by factors of 5-100 on a single processor over unoptimized simulations without introducing additional approximations. The speedup for a particular surface depends on the self-similarity of the surface at the scale of the electron penetration depth. We further accelerate by parallelizing the calculations for a total speedup of about 100-2000 on 30 processors. The goal of this work was to create a system capable of simulating a quantitatively accurate SEM image of a relatively unconstrained surface. Results of this work include simulation software, optimization algorithms, performance measurements with various optimizations, and examples of simulated images. PMID- 14748391 TI - Scanning the industry. Detailing the evidence. PMID- 14748392 TI - [Spontaneous regression of testicular tumor]. AB - We report the case of a 33-year-old man presenting with a retroperitoneal seminoma secondary to a testicular tumor which had completely and spontaneously disappeared (burned-out or shrinking seminoma). Ultrasound only showed a fibrous scar in the right testis. A careful study of the testicles should be considered in all male retroperitoneal tumor, even if they are asymptomatic, to exclude an occult cancer or more rarely a spontaneously involuted tumor. Testicular ultrasonography is today the gold-standard technique and any parenchymal echostructure abnormality should be verified histologically. PMID- 14748393 TI - Rotary fixation of the atlantoaxial joint. AB - Rotary fixation of the atlanto-axial joint is a rare disorder in which the atlas gets fixed in a position normally achieved during rotation. A persistent asymmetrical position of the dens in relation to the atlas during head rotation is the clue to the diagnosis. Rotary fixation can be demonstrated by radiography/fluoroscopy and computed tomography (CT). However it is easily overlooked, largely by inattention. We present a typical case in an adult in whom the rotary fixation was demonstrated by radiography/fluoroscopy and CT. PMID- 14748394 TI - Selective arterial embolization in the treatment of an aneurysmal bone cyst of the pelvis. AB - This case report deals with an aggressive aneurysmal bone cyst of the pelvis in a 20-year-old man causing considerable destruction of the left ilium and the anterior column of the acetabulum, with extension into the pelvis. The treatment of large pelvic aneurysmal bone cysts is challenging because of local destruction of adjacent structures and because of the risk of severe intraoperative bleeding. Therefore selective preoperative embolization may be a valuable tool in the management of aneurysmal bone cysts. PMID- 14748395 TI - An unusual case of hydatid disease located in the erector spinae muscle. AB - We present a case of hydatid disease localized to the erector spinae muscle. A 60 year-old male was admitted to our hospital with back pain. The diagnosis was made preoperatively by US, CT, and MRI. They showed hydatid disease in erector spinae muscle. Cysts and some part of the erector spinae muscle were resected. Postoperatively pathology confirmed the diagnosis. PMID- 14748396 TI - Imaging of semimembranosus bursitis: MR findings in three patients and anatomical study. AB - We report on three patients with posteromedial knee pain related to semimembranosus bursitis. The semimembranosus bursa has a typical location and morphology around the central tendon of the semimembranosus muscle. These characteristics are demonstrated in patients and by dissection of the tendon and bursa in an embalmed knee specimen. PMID- 14748397 TI - CT angiography: basic principles and post-processing applications. AB - The introduction of spiral and more specifically multidetector CT scanners has provided unique opportunities in the field of CT angiography. It has also stressed the need of a basic knowledge of post-processing 3D imaging techniques to obtain maximum information of the CT data set. Aim of this article is to give a basic review of technical issues concerning CT angiography, with special attention to 3D imaging techniques such as surface shaded display (SSD), maximum intensity projection (MIP) and volume rendering (VR). PMID- 14748398 TI - Phase-contrast magnetic resonance angiography. AB - Time of flight (TOF) and phase contrast (PC) magnetic resonance angiography (MRA) are two established magnetic resonance (MR) techniques that were described and developed in the 1980s. Both of these techniques usually do not involve the use of intravascular contrast, but rather exploit the intrinsic contrast provided by flowing blood to produce vascular signal. Bolus infused (gadolinium-enhanced) MR angiography was only introduced in 1993. PC-MRA uses the phase shifts introduced to nuclei with motion in the presence of a magnetic field gradient. A bipolar magnetic field gradient will induce a phase shift to nuclei moving along the gradient dependent on the velocity, as well as acceleration and higher order motion terms. More complex gradient waveforms enable sensitivity to specific motion terms such as velocity or acceleration. By constructing an image in which the intensity is proportional to the phase shift of the nuclei, it is possible to create an angiographic image related to the flow properties of blood (or other liquids such as cerebrospinal fluid). The PC-MRA is a powerful technique and allows for encoding of flow in one or many directions in such a way that the velocity sensitivity can be chosen depending on the vessel of interest. This technique also allows for quantification of flow velocity and flow rate, which is not generally available with other angiographic techniques. PMID- 14748399 TI - Basics of non-invasive angiography contrast-enhanced magnetic resonance angiography. AB - Contrast-enhanced magnetic resonance angiography is a non-invasive imaging modality without catheterization or nephrotoxicity. This is accomplished by using paramagnetic contrast agents in combination with very rapid acquisition sequences. Paramagnetic contrast agent is injected intravenously and image data are collected during the first-pass of contrast agent through the vascular territory of interest. Due to the strong enhancement effect of paramagnetic contrast agents, a small dose injected as an intravenous bolus is sufficient to briefly enhance the entire arterial vascular tree. This allows imaging with a large field-of view that encompasses an extensive region of vascular anatomy. By using a dedicated 3D imaging sequence on scanners with high-performance gradient systems, high-resolution 3D volumes of image data can be acquired in a single breath-hold. This has vastly improved image quality of 3D contrast-enhanced MRA (CE-MRA) exams, particularly in the chest and abdomen. Subsequent postprocessing allows an angiographic display of image data in any desired obliquity. The following part gives an overview and the basic principles of the methods used for CE-MRA. PMID- 14748400 TI - Principles of time-resolved MRDSA. AB - Time resolved Magnetic Resonance Digital Subtraction Angiography (MRDSA) is introduced in the context of contrast enhanced MRA as a fast technique implementable in 2D or 3D versions. Particularities of the technique are briefly discussed. Sense Time Resolved MRDSA is presented and clinical examples are given. PMID- 14748401 TI - Imaging strategy for acute stroke therapy. AB - New MR imaging methods provide us with a hitherto unimaginable wealth of information on the pathophysiologic mechanisms operative in acute stroke. The only evidence-based acute stroke therapy, thrombolysis with IV tPA, only requires a CT to exclude brain hemorrhage before treatment. In PROACT 2 (Prolyse in Acute Cerebral Thromboembolism) locoregional thrombolysis with prourokinase improved the frequency of independent outcome. The only imaging methods used in this trial were a brain CT to exclude patients with brain hemorrhage and extended early infarct signs and a classical angiogram to demonstrate the presence of an M1 or M2 segment occlusions. In principle these therapies can be applied without using sophisticated MR or CT techniques. In this brief review we discuss the potential role of these novel techniques before application of intravenous or intraarterial thrombolysis. PMID- 14748402 TI - MR angiography in steno-occlusive disease of the intracranial vessels. AB - Technical advances in MR angiography (MRA) have improved the sensitivity and the accuracy of this technique in the evaluation of vascular stenoocclusive disease of the brain. In many centers, due to its non-invasive and non-ionizing character, MR angiography has already replaced conventional angiography for the screening of intracranial vascular disease. Several MRA techniques have been developed for imaging the intracranial vascular system, such as time-of-flight MRA (TOF-MRA), phased-contrast MRA (PC-MRA), and more recently contrast-enhanced MRA (CE-MRA). PMID- 14748403 TI - Supraaortic vessels: a clinician's point of view. AB - Stroke has been recognized as the third leading cause of death in the Western World, and more than 80% of all ischemic events is caused by arteriosclerosis affecting the extracranial arteries, especially the carotid bifurcation. Carotid endarterectomy (CEA) is the gold standard for preventing stroke in symptomatic and asymptomatic patients with significant carotid stenosis, when performed in centers with low complication rates (less than 3%). Recently, the carotid artery stenting (CAS) has been proposed as an alternative procedure, although its safety compared to conventional CEA is not fully documented yet. For both techniques, an accurate diagnosis by various imaging techniques is very important regarding the risk of serious complications following surgical intervention. PMID- 14748404 TI - Images in clinical radiology. Small bowel adenocarcinoma in a 28-year-old patient. PMID- 14748405 TI - Images in clinical radiology. Glomus tympanicum: CT diagnosis. PMID- 14748407 TI - Changes in the hemodynamic state of patients with acute lethal organophosphate poisoning. AB - The main cause of death due to acute organophosphate poisoning is believed acute respiratory failure caused by peripheral and central cholinergic actions. Today, advances in respiratory management and intensive care make it possible to maintain the respiratory function of patients with organophosphate poisoning, but it is still difficult to maintain their circulation, and some patients with acute organophosphate poisoning die of metabolic acidosis. The present study clarified the hemodynamics of patients with acute lethal organophosphate poisoning. Subjects were patients with severe acute organophosphate poisoning accompanied by hypotension in whom hemodynamics were monitored by pulmonary artery catheterization. In this study, the medical records of these patients were reviewed. Four patients with severe acute organophosphate poisoning accompanied by hypotension and metabolic acidosis died. In 3/4 patients respiration was maintained favorably. In all 4 patients cardiac output was maintained, but systemic vascular resistance index (SVRI) was significantly reduced. Catecholamine administration was ineffective and did not increase SVRI. PMID- 14748406 TI - Antioxidant and neuroprotective activity of the extract from the seaweed, Halimeda incrassata (Ellis) Lamouroux, against in vitro and in vivo toxicity induced by methyl-mercury. AB - A growing body of evidence suggests oxidative stress as part of the toxicity mechanism of methyl-mercury (MeHg) in cell cultures and animal models and so justifies the use of natural antioxidants as therapeutic alternatives. This research examines the effect of an aqueous extract from the marine seaweed Halimeda incrassata (Hi) against the oxidative stress induced by MeHg on in vitro and in vivo models. In GT1-7 mouse hypothalamic cell cultures, the extract of Hi increased cell viability and reduced ROS production after 24-h exposure to MeHgCl. Wistar rats, acutely intoxicated with MeHgCl, had reduced levels of serum and brain thiobarbituric reactive substances when treated with the Hi extract. Similarly, animals exposed to repeated doses of MeHgCl were protected by the seaweed extract from variations in body weight, food consumption and the appearance of neurological effects. This research supports the notion that oxidative stress is directly involved in MeHg intoxication, so that natural antioxidants, particularly those in the extract of Hi, can be useful therapeutic alternatives. PMID- 14748408 TI - Neurotoxic, hemorrhagic and proteolytic activities of Duvernoy's gland secretion from Venezuelan opisthoglyphous colubrid snakes in mice. AB - Many colubrid snakes produce toxic oral secretions. We studied venom (Duvernoy's gland secretion) collected from Venezuelan opisthoglyphous (rear-fanged) colubrid snakes. Different proteins were present in Thamnodynastes stigilis Duvernoy's gland secretion and were characterized by 20% SDS-PAGE protein electrophoresis. The venom displayed proteolytic (gelatinase) activity that was partially purified on a chromatography ionic exchange mono Q2 column. We demonstrated hemorrhagic activity of Thamnodynastes stigilis Duvernoy's gland secretion on chicken embryos and mouse skin and peritoneum. Mice inoculated with Thamnodynastes stigilis Duvernoy's gland secretion presented signs of neurotoxicity. Thamnodynastes stigilis Duvernoy's gland secretion had proteolytic, hemorrhagic, and neurotoxic activities, not previously described in this species and identifies the presence of a new venomous colubrid in Venezuela. PMID- 14748409 TI - In vitro reactivation of acetylcholinesterase using the oxime K027. AB - The ability of a new bisquaternary oxime, K027 (1-[4 hydroxyiminomethylpyridinium]-3-[carbamoylpyridinium] propane dibromide), to reactivate the enzyme acetylcholinesterase (AChE) inhibited by the nerve agents Tabun, sarin and VX was evaluated. Its reactivation potency was compared to the AChE reactivators pralidoxime (2-PAM), obidoxime and HI-6; K027 seems a good reactivator of organophosphates-inhibited AChE. Its reactivation potency is lower compared to the other oximes for reactivation of sarin-inhibited AChE, but it is sufficient to significantly increase the activity of sarin-inhibited AChE. Its reactivation ability is comparable to obidoxime for reactivation of VX- and tabun inhibited AChE and is higher than the reactivation potency of HI-6, for tabun inhibited AChE. HI-6 is currently regarded the most promising reactivator of organophosphates-inhibited AChE. PMID- 14748410 TI - Two hundred and thirteen cases of marijuana toxicoses in dogs. AB - Marijuana (Cannabis sativa) is a commonly used recreational drug among humans; animals may be exposed following ingestion or accidental inhalation of smoke. From January 1998 to January 2002, 213 incidences were recorded of dogs that developed clinical signs following oral exposure to marijuana, with 99% having neurologic signs, and 30% exhibiting gastrointestional signs. The marijuana ingested ranged from 1/2 to 90 g. The lowest dose at which signs occurred was 84.7 mg/kg and the highest reported dose was 26.8 g/kg. Onset of signs ranged from 5 min to 96 h, with most signs occurring within 1 to 3 h after ingestion. The signs lasted from 30 min to 96 h. Management consisted of decontamination, sedation (with diazepam as drug of choice), fluid therapy, thermoregulation and general supportive care. All followed animals made full recoveries. PMID- 14748411 TI - Association between droperidol use and sudden death in two patients intoxicated with illicit stimulant drugs. AB - Illicit drug intoxication is often a cause of extreme agitation in the emergency department and prehospital settings. Chemical restraint is often required to protect patient as well as health-care providers. Droperidol has commonly been used to sedate extremely agitated patients in the emergency department and psychiatric settings. Its safety has been demonstrated in these settings and in patients who's agitation has been attributed to amphetamine toxicity. We present 2 cases of sudden death following the use of droperidol to sedate 1 patient who was extremely agitated secondary to cocaine intoxication and another secondary to phencyclidine intoxication. PMID- 14748412 TI - Acute neuropathy and erythromelalgia following topical exposure to isopropanol. AB - Adverse effects resulting from topical exposure to isopropanol appears exceptional in adults with intact skin. We describe the case of a young woman who developed an acute sensori-motor axonal polyneuropathy after walking bare-feet for several hours on carpets soaked by a disinfectant containing isopropanolol. The persistence and severity of symptoms raised the possibility of her neuropathy being partly related to immunizations she received 1 mo earlier. The occurrence shortly after contact, however, strongly suggested responsibility of the dermal isopropanol exposure. This case being, to our knowledge, the second reported, peripheral nerve toxicity appears possible in adults on prolonged topical exposure, probably in susceptible individuals. PMID- 14748413 TI - Salt poisoning in beef cattle. AB - Salt poisoning has been described under various circumstances in adult cattle. Presenting clinical signs in 6 Holstein beef cattle with such poisoning were primarily dysfunction of the central nervous system and included ataxia, opisthotonus, nystagmus, depression, muscle twitching and intermittent convulsions, as well as abdominal pain and polydipsia. Diarrhea occurred in 2, and blindness in 3/6 cattle. Hypernatremia (161.8 - 178.8 mmol/L) and hyperosmolality (331.81 - 366.18 mOsm/L) were present in all animals. To treat the affected cattle, access to fresh water was restricted, vascular volume was expanded with isotonic saline and then hypotonic fluid (5% Dextrose solution) i.v. and dexamethasone im was administered. Although biochemical parameters returned to normal reference ranges, 3/6 affected animals remained blind. PMID- 14748414 TI - Use of neostigmine in massive ivermectin toxicity in cats. AB - Over-the-counter availability of drugs in developing countries poses the risk of their abuse and overdosing in animals by the owners. This report describes ivermectin toxicosis and its treatment with neostigmine methylsulfate in 2 kittens and an adult cat that had been massively overdosed by their owners. Of two 4-w-old 300 g kittens, one was injected with 1.5 ml ivermectin (15 mg; corresponding to 250 times the recommended dose) by the owner and became comatose, started salivation and died after 2 1/2 h. The other kitten was treated with about 0.75 ml ivermectin (7.5 mg; corresponding to 125 times the recommended dose) by the same owner and developed ataxia and salivation after 2 h, went into coma the next morning and was presented for treatment with mydriasis, coma, slight fever (102.4 F), tachypnea (48/min) and severe tachycardia. This kitten was treated twice at a half hourly interval with neostigmine methylsulfate (25 microg) and 5% dextrose (20 ml) iv showed a transient improvement but died 12 h later. A 2-y-old, 4.5 kg male cat was treated s.c. with 1.5 ml ivermectin (15 mg; corresponding to 16.6 times the recommended dose) by its owner and developed slight salivation, lacrimation. mydriasis, protrusion of third eye-lid, tachypnea (42 breaths/min), tachycardia (128 beats/min) and ataxia. The patient was treated with neostigmine methylsulfate (150 microg) and 5% dextrose (100 ml) i.v., and 200 ml of Heartmann's solution i.v. Treatment was repeated after 6 h Follow up treatment over next 2 d was with daily i.v. administration of neostigmine and dextrose. The patient completely recovered 5 d after initiation of treatment. PMID- 14748415 TI - The fentanyl tea bag. AB - Fentanyl patches create unique opportunities for use and abuse. Each patch contains 100-fold more drug than is stated on the label in order to create the gradient required to deliver the stated amount (ie 25-100 microg/h). Several methods of abuse of this analgesic have been reported, ranging from ingestion to inhalation to application of multiple patches to the skin. We report the unique case of a 21-y-old woman who steeped a fentanyl patch in a cup of hot water and then drank the mixture. Coma and hypoventilation resulted. The woman was resuscitated with naloxone i.v. and recovered without sequelae. PMID- 14748416 TI - Poisonous plants in the eastern Mediterranean basin. AB - Plants found in the eastern Mediterranean are discussed by their toxic principles and poisonous effects. Nitrate- and oxalate-containing plants, those with alkaloids and glycosides, plants inducing photosensitization and carrying tannins, and selected miscellaneous species of toxic plants are reviewed and their poisonous properties described. PMID- 14748417 TI - Trends in age-specific human poisoning exposures reported to a regional poison control center, 1997-2001. AB - This study examined age-specific trends in the annual incidence rates of poisoning exposures reported to a regional poison control center over a 5-y period. Data from the poison control center's 1997-2001 computerized case records were combined with age-specific population estimates from the US Census Bureau. Incidence rates of reported poisoning exposures were calculated and evaluated for the following age groups: < 6 y; 6 to 12 y; 13 to 19 y; 20 to 35 y; 36 to 65 y; and > 65 years. Ordinary linear regression analysis revealed a statistically significant decreasing trend in the annual crude incidence rate of poisoning exposures/1.000 population (r2 = .87, slope = -.33 p = .02). Different patterns were observed in the rates of reported poisoning exposures between the 6 age groups. While there was a significant decrease in the rate of reported poisoning exposures in children < 6 y of age or less, and for adults 20-35 and 36-65 y-old, the incidence rate among children 6-12 y, adolescents, and people > 65 y-old showed no significant decreasing trend. PMID- 14748418 TI - Significant changes in antipsychotic drug use during pregnancy. AB - Atypical antipsychotics are less likely to cause hyperprolactinemia-related side effects, such as infertility; hence it is predicted that more women taking antipsychotic medications will be able to become pregnant as the use of atypical antipsychotics increases. To compare the use of conventional and atypical antipsychotics, we conducted a retrospective review of the Motherisk Program clinic schedule from 1989 to 2001 comparing the proportion of appointments made for conventional and atypical antipsychotics. In 1989, 2.7% of all appointments were about the use of antipsychotic medication. In 2001, 7.4% of appointments were regarding antipsychotic drug use. This 170% increase was due to an increase in appointments for atypical antipsychotics as the number of appointments for conventional antipsychotics remained relatively constant over the 12-y period. Since the introduction of atypical antipsychotics, more women requiring antipsychotic drug therapy have been planning or becoming pregnant. This increase may have substantial public health implications. PMID- 14748419 TI - Two years' activity of the Veterinary Toxicology Attention Service of Lugo, Spain. AB - The Veterinary Toxicology Attention Service was created at the beginning of 2001 as the first on-line toxicology service for veterinarians and animal owners in Spain. In the present study, data about the general functioning of the Service and the toxicological analysis and consultations performed are summarized. Canine related cases constituted the main call group and veterinary practitioners represented half of the consultations. Coordination between all veterinary toxicology services in Spain and the rest of the European Union should improve this service. PMID- 14748420 TI - Using and developing your EDTNA/ERCA Professional Portfolio. AB - A portfolio is an instrument for continuing professional development, which can be used by all members of the multidisciplinary team. This paper describes the development of the EDTN/ERCA Professional Portfolio and discusses how health care professionals are able to complete and utilise the Portfolio in everyday practice. A discussion on reflective practice is presented and a short example of reflective writing is given. The paper concludes with an evaluation of possible Portfolio use in teaching and management and discusses plans for future development. PMID- 14748421 TI - The European Basic Core Curriculum for Nephrology Nursing, the process of writing, publishing and disseminating. AB - In August 2002 the European Basic Core Curriculum (BCC) for Nephrology Nursing was published on the EDTNA/ERCA homepage. At the EDTNA/ERCA annual conference in The Hague in September 2002, it was presented to the delegates as an oral presentation and a poster presentation. Publicity flyers were translated into seven languages and distributed to delegates. This paper will describe the aims of the BCC, the process of writing and publishing, the use of the BCC in nursing schools and the measures that have already been taken in implementation and dissemination. PMID- 14748422 TI - Accreditation of post-basic nephrology courses within the EDTNA/ERCA. AB - This paper will outline the development of the EDTNA/ERCA accreditation programme for post-basic courses in nephrology. It will discuss results of the pilot project and relate the experiences of one school that has successfully gone through the accreditation process. The article will conclude with a discussion on more recent developments and evaluate the project in light of the original concepts. PMID- 14748423 TI - Reflections on issues related to specialist nursing education. PMID- 14748424 TI - Recommendations for a European framework for specialist nursing education. AB - Whereas in many countries within the EU specialist nursing practice exists requiring post-basic * nursing education that varies from one country to another, the European Network for Nurses Organisations has developed a framework for specialist nursing education, in order to harmonise post basic nursing education and to facilitate the free movement of specialist nurses. PMID- 14748425 TI - A university accredited renal nursing course delivered by distance learning. AB - Continuous professional development (CPD) is limited in some regions of the UK and within Europe generally. This is compounded for all by limited resources for course fees and the lack of study leave granted away from the clinical area for full-time courses. This is set against recommendations from national and European governments and renal clinical guidelines concerning expectations of CPD and competency levels of renal nurses. In the past renal nurses have been trained in all areas of the speciality by local Schools of Nursing linked to renal units. However, since the formation of Trusts in 1990 education delivery has been ring fenced and separated from the control of local hospitals by Schools of Nursing being incorporated into Institutes of Higher Education. That has led in some instances to a rationalising of post registration education delivery for some geographical areas. This paper will discuss the development and implementation of a distance learning renal care course taking into consideration the clinical, academic and educational requirements necessary for effective education and training at the post basic level. PMID- 14748426 TI - Discovering the culture of haemodialysis units: an ethnography. AB - There is an increasing renal population but a lack of resources available to ensure that every renal patient receives the maximum benefit from Renal Replacement Therapy (RRT). The aim of this paper is to illustrate how understanding the culture of dialysis units will facilitate the provision of more therapeutic, patient centred care. This paper describes briefly the research study that was undertaken to uncover the culture which is in dialysis units but as yet undiscovered and therefore not valued. Once this culture is better understood and valued both patients and the health care teams will be facilitated to work towards the common goal of better treatment outcomes. PMID- 14748428 TI - A case study--initiation of haemodialysis. AB - End-stage renal disease (ESRD) is a chronic failure of normal kidney function, which is precipitated by a number of factors, such as diabetes, pyelonephritis, hypertension, and polycystic kidney disease (1). The loss of kidney function is gradual, and many patients do not have any symptoms until the end-stage of the disease. Killingworth and Van Den Akker (2) point out that people undergoing haemodialysis have to comply with a wide range of medications and restrictions, which affect both their physiological and psychological status. This paper presents a case study of a woman with ESRD, requiring initiation of haemodialysis. An assessment of the patient's health needs is addressed, followed by the implementation and evaluation of the nursing process. Finally, the analysis of the overall plan used in the case study is presented. PMID- 14748427 TI - Education to reduce potassium levels in adolescent, haemodialysis patients. AB - A semi-experimental survey was carried out. The aims included assessing how knowledgeable adolescents undergoing haemodialysis treatment were regarding hyperkalaemia, and determining how effective an education programme in preventing hyperkalaemia would be. 31 volunteers (twelve girls and nineteen boys), who had two or three haemodialysis treatments a week, were included in the study. Data were obtained through a questionnaire, the answers to which were written down for each participant separately by the researcher. A manual was prepared with the aim of educating children in hyperkalaemia, the participants were taught lessons in groups of two to three at a time. The education programme was completed in eight sessions. At the end of the programme each participant was given a copy of the manual. A month after the education programme, participants were asked to refill the section of the aforesaid questionnaire. The scores in this section had increased, and blood potassium levels had significantly decreased (p = 0.02). PMID- 14748429 TI - A nurse led central line insertion service. AB - Advanced Nurse Practitioners (ANP) are developing their role in order to insert temporary and permanent central venous catheters to facilitate haemodialysis. Two hundred and eighty-nine central venous catheters (CVC) were inserted over a two year period. The ANP inserted 117 CVC with 172 CVC inserted by the nephrology medical team. Using non-parametric independent two-group comparison Mann Whitney test and Chi-Squared test, the two groups were compared for specific outcomes. When comparing both groups with respect to CVC longevity, elective and non elective removal along with infection (exit site and systemic) no statistical difference between the ANP and the nephrology medical staff could be found. PMID- 14748430 TI - The hurdles of migrant nurses. AB - The vast disequilibrium between healthcare providers and the recipients of care creates a hazard effect to all. This is a global issue as who will look after the growing and ageing population? Some countries have decided to recruit from outside their own country. Prior to registration, the Board of Nursing has imposed a regulation that a foreign nurse has to undergo a three- to six-month adaptation programme. The main objective is to assess and evaluate and at the end of the programme it is hoped that the nurse will be deemed competent and safe to practice. Foreign nurses face tremendous challenges but are able to integrate with the provision of support services and an adaptation programme. This programme needs to be culturally sensitive to nurses who are used to work in a different system of health care. PMID- 14748431 TI - Burnout in renal care professionals. AB - Burnout is defined as "a syndrome of emotional exhaustion, depersonalisation and reduced personal accomplishment that can occur among individuals who work with people in some capacity", and it can be considered as a result of long-term exposure to occupational stress. Frequently reported occupational stressors among caring professionals are those intrinsic to the job, related to patient demands, related to roles within the organisation, and those related to relationships at work and career development. In renal care however, there are some unique characteristics such as technologically advanced equipment, the intensive caring environment and the long-term relationships being established between the carer and chronic renal patients, that one should take into consideration. It seems that job resources may act as moderators to burnout. Furthermore, specific personality characteristics and socio-demographic variables seem to affect the burnout experience. Individual and social organisational means for burnout prevention and coping are discussed and suggested. PMID- 14748432 TI - Needling fistulas bevel down. PMID- 14748433 TI - Iron management in renal failure patients--how do we achieve the best results? PMID- 14748434 TI - The use of housekeeping genes (HKG) as an internal control for the detection of gene expression by quantitative real-time RT-PCR. AB - Quantitative real-time RT-PCR is a very useful technique for estimating gene expression at the mRNA level. The expression of a tested gene has to be compared with that of a control gene. Various housekeeping genes have been used as control genes in different systems. In our study we tested several housekeeping genes in the model of gene expression after induction of apoptosis and differentiation. The myeloid cell lines were incubated with phorbol esters, butyric acid and combination of TNFalpha and IFNgamma to induce differentiation. Camptothecin was used for induction of apoptosis. Tested control genes included beta2 microglobulin, GAPDH, 18S ribosomal RNA and abl. GAPDH was found to be the best control gene in the apoptotic system. Different control genes were suitable for different systems where differentiation or senescence was induced. Our results show that attention should be paid to the choice of an appropriate control gene of quantitative real-time RT-PCR for different experimental models and various experimental conditions. PMID- 14748435 TI - Adjuvant cytokine treatment of minimal residual disease after surgical therapy in mice carrying HPV16-associated tumours: cytolytic activity of spleen cells from tumour regressors. AB - It has been found previously that IL-2, IFNgamma and GM-CSF were capable of reducing the recurrence rate of HPV 16-associated tumours in mice with SMRTD. We were interested whether the therapeutic effect of the surgery and adjuvant cytokine treatment was accompanied by cytolytic activity of spleen cells and whether the activity of the spleen cells was different in mice that had rejected tumour residua after surgery and adjuvant therapy with cytokines (tumour regressors) as compared to those that had not rejected the tumour residua (tumour progressors). We have examined the cytolytic activity of spleen cells from MHC class I+ TC-1 tumour regressors and progressors after treatment of TC-1 SMRTD with GM-CSF, and the activity of spleen cells from MHC class I- MK16 tumour regressors and progressors after treatment of MK16 SMRTD with IL-2 and IFNgamma. It has been found that irrespective of the tumour type and adjuvant treatment, the spleen cells from tumour regressors after surgery were regularly more cytolytic when allowed to react with target cells from HPV 16-associated tumours than the spleen cells from tumour progressors. No substantial differences between the cytolytic activity of spleen cells from the operated-only and operated plus cytokine (GM-CSF, IL-2, IFNgamma) adjuvant treated groups were observed. The cytolytic activity of spleen cells from mice with SMRTD allowed to react with MHC class I+ , MHC class I-, NK-sensitive and NK-resistant targets is compatible with the interpretation that in the mice with MHC class I+ TC-1 tumours, primarily cytotoxic T lymphocytes (CTL) were efficient, whereas in the mice with MHC class I- MK16 tumours, both NK and non-lymphocytic effector cells were involved. PMID- 14748436 TI - Differential linkage of triglyceride and glucose levels on rat chromosome 4 in two segregating rat populations. AB - The PD/Cub is a recently established model of the IRS. The BN.SHR4 congenic strain was derived by introgression of the chromosome 4 segment of SHR origin (including the defective Cd36/Fat allele) onto the BN/Cub genetic background. We investigated the linkage of metabolic and morphometric phenotypes (total body weight, OGTT, fasting serum levels of TG, FFA) on chromosome 4 in two separate F2 rat populations: the PD/Cub x BN/Cub and PD/Cub x BN.SHR4 (total N = 243). In the PD/Cub x BN.SHR4 F2s, we found significant linkage for fasting TG levels (LOD = 3.26) and suggestive linkage for fasting glycaemia (LOD = 2.80) in the interval Il-6 - D4Bro1, i.e. the part of chromosome 4 of SHR origin in the BN.SHR4 congenic. However, no linkage for fasting TG concentrations, fasting glycaemia or any other followed parameter was found in the second, PD/Cub x BN/Cub F2. The differential linkage of TG and glucose levels to the centromeric part of rat chromosome 4q in the studied F2s points to the importance of this region for the lipid and carbohydrate metabolism at the specific age (10 months) and diet (standard chow) combination. The Cd36/Fat and Il-6 genes are the preliminary positional candidates for the observed effect. PMID- 14748437 TI - Identification of HLA alleles with low or no cell surface expression in the Czech population. AB - The presence of the A*24020102L allele is implicated in one donor from the CBMD who serologically was typed as A2; B44, B55; Cwl, Cw7. The DRB4*01030102N allele was identified in one healthy donor and in one patient with MDS during routine HLA class II DNA typing. The DRB4*01030102N allele was identified in the patient's father, who had CML, and was associated with the HLA-A3-B7-Cw7 DRB1*0701-DQB1*0303 haplotype, which is common for European populations. In order to avoid mistyping, both techniques, serology and molecular biology must be used for HLA typing, especially for cases where just one antigen appeared to be present using serological methods. PMID- 14748438 TI - MHC class I+ and class I- HPV16-associated tumours expressing the E7 oncoprotein do not cross-react in immunization/challenge experiments. AB - It has been demonstrated repeatedly that a high proportion of tumours derived from MHC class I+ precursors are MHC class I-. Since a major task in immunotherapy strategies for treatment of malignancies is to develop polyvalent tumour vaccines efficient against a broad spectrum of tumours, we have examined whether MHC class I+ cell-based tumour vaccines can cross-protect against homologous MHC class I- tumour challenge and vice versa. For these purposes, we have used two oncogenic cell lines induced independently by co-transfection of murine H-2b cells with E61E7 HPV16 and activated Ha-ras oncogenes, the tumours TC 1 (MHC class I+, HPV16 E7+) and E7+). Surprisingly, it was found that these two tumours do not cross-react, although both of them contain the crucial HPV16-coded tumour rejection antigen E7. Preimmunization with the MHC class I+ tumour did not protect against a subsequent challenge with the MHC class I- tumour and vice versa; however, immunization with the TC-1 tumour could protect syngeneic mice against the TC-1 tumour challenge and, similarly, immunization with the MK16/1/IIIABC tumour could protect mice against the MK16/1/IIIABC tumour challenge. If this finding can also be confirmed as a more general phenomenon with other MHC class I+ and class 1- tumours, it could have serious implications for design of immunotherapeutic vaccines and protocols. PMID- 14748439 TI - No association between oxidative stress and IL-6 in NIDDM patients with nephropathy. AB - Oxidative stress is hypothesized to play a role in the development of diabetes with and without nephropathy. In addition, it has been suggested that some metabolic abormalities associated with diabetes may be due to cytokine overproduction. In the light of this knowledge, we aimed to measure MDA levels as a marker of oxidative stress and the IL-6 level in diabetes with and without different stages of nephropathy. Plasma MDA levels in the group of NIDDM patients with advanced nephropathy were significantly higher than in the group of NIDDM patients without nephropathy, which had significantly higher levels compared with the control group. Although IL-6 levels were elevated in diabetic groups with and without nephropathy in comparison with the control, no significant difference was found between patient groups. As a conclusion, oxidative stress may play an important role in diabetes with and without nephropathy, but the IL-6 level may not be useful in the evaluation of diabetic nephropathy. PMID- 14748440 TI - Monoclonal antibody BF-06 against the heavy chain of clathrin. PMID- 14748441 TI - Phonological acquisition: recent attainments and new challenges. AB - Infants' phonological acquisition during the first 18 months of life has been some 30 years. Current research themes include statistical learning mechanisms, early lexical development, and models of phonetic category perception. So far, linguistic theories have hardly been taken into account. These theories are based upon the assumption that there is a common core of innate phonological knowledge across speakers of all human languages, and they contain detailed proposals concerning phonological representations and the derivations by which abstract underlying forms are mapped onto concrete surface forms. It remains to be investigated experimentally if there is innate phonological knowledge and how the language-specific phonological grammar is acquired. In the present article, the contributions to this special issue are introduced, and an attempt is made to bridge the gap between phonological theory and experimental psychology. In particular, some recent experimental work is considered in the light of phonological theories and new research avenues are sketched. What might be innate, what needs to be acquired, and how this acquisition might take place are questions that are addressed with respect to several aspects of phonological knowledge, specifically segmental representations, phonotactics, phonological processes, and the architecture of the phonological grammar. PMID- 14748442 TI - Phonetic diversity, statistical learning, and acquisition of phonology. AB - In learning to perceive and produce speech, children master complex language specific patterns. Daunting language-specific variation is found both in the segmental domain and in the domain of prosody and intonation. This article reviews the challenges posed by results in phonetic typology and sociolinguistics for the theory of language acquisition. It argues that categories are initiated bottom-up from statistical modes in use of the phonetic space, and sketches how exemplar theory can be used to model the updating of categories once they are initiated. It also argues that bottom-up initiation of categories is successful thanks to the perception-production loop operating in the speech community. The behavior of this loop means that the superficial statistical properties of speech available to the infant indirectly reflect the contrastiveness and discriminability of categories in the adult grammar. The article also argues that the developing system is refined using internal feedback from type statistics over the lexicon, once the lexicon is well-developed. The application of type statistics to a system initiated with surface statistics does not cause a fundamental reorganization of the system. Instead, it exploits confluences across levels of representation which characterize human language and make bootstrapping possible. PMID- 14748443 TI - A statistical basis for speech sound discrimination. AB - Infants under six months are able to discriminate native and non-native consonant contrasts equally well, but as they learn the phonological systems of their native language, this ability declines. Current explanations of this phenomenon agree that the decline in discrimination ability is linked to the formation of native-language phonemic categories. The goal of this study was to evaluate the role of input statistics in learning these categories: our hypothesis was that relative frequency is a determinant of the relative order in which categories are acquired. English-learning infants of two age groups (6.5 months and 8.5 months) were tested on their ability to discriminate non-native consonant contrasts using the Conditioned Head Turn Procedure. As predicted, older infants were worse in their performance on the more frequent coronal stop contrast than on the less frequent dorsal stop contrast. In contrast, 6.5-month-olds discriminated both contrasts equally well. An adult control group tested with an AX task also discriminated both contrasts equally. These results provide preliminary confirmation of the hypothesis that frequency plays an important role in tuning of phonological systems to properties of the native language. A simple attractor model suffices to account for these and previous results on loss of discrimination of non-native-language contrasts and suggests that the technique of measuring graded loss of multiple contrasts, in combination with observation of input frequencies, can offer a powerful method of assessing infants' phonological representations. PMID- 14748445 TI - Simultaneous bilingualism and the perception of a language-specific vowel contrast in the first year of life. AB - Behavioral studies have shown that while young infants can discriminate many different phonetic contrasts, a shift from a language-general to a language specific pattern of discrimination is found during the second semester of life, beginning earlier for vowels than for consonants. This age-related decline in sensitivity to perceive non-native contrasts has been generally attested in monolinguals. In order to analyze the impact of bilingual exposure on the perception of native-sound contrasts and the early building of language-specific contrastive categories, four-month-old and eight-month-old infants from Spanish monolingual, Catalan monolingual and Spanish-Catalan bilingual environments have been tested with a familiarization-preference procedure on a vowel contrast present only in Catalan: /e/-/epsilon/. As expected, younger infants were all able to perceive this contrast, independently of the language of exposure. However, by eight months, only infants from Catalan monolingual environments succeeded. Although the decline in sensitivity with the monolingual Spanish group was expected, the results with the bilingual group challenge the view that mere exposure is enough to maintain the capacity to perceive a contrast. An additional experiment at 12 months of age indicated that bilinguals finally regained discrimination. Together these results suggest a specific developmental pattern of perceptual reorganization in bilingual exposure. PMID- 14748444 TI - Infant perception of non-native consonant contrasts that adults assimilate in different ways. AB - Numerous findings suggest that non-native speech perception undergoes dramatic changes before the infant's first birthday. Yet the nature and cause of these changes remain uncertain. We evaluated the predictions of several theoretical accounts of developmental change in infants' perception of non-native consonant contrasts. Experiment 1 assessed English-learning infants' discrimination of three isiZulu distinctions that American adults had categorized and discriminated quite differently, consistent with the Perceptual Assimilation Model (PAM: Best, 1995; Best et al., 1988). All involved a distinction employing a single articulatory organ, in this case the larynx. Consistent with all theoretical accounts, 6-8 month olds discriminated all contrasts. However, 10-12 month olds performed more poorly on each, consistent with the Articulatory-Organ-matching hypothesis (AO) derived from PAM and Articulatory Phonology (Studdert-Kennedy & Goldstein, 2003), specifically that older infants should show a decline for non native distinctions involving a single articulatory organ. However, the results may also be open to other interpretations. The converse AO hypothesis, that non native between-organ distinctions will remain highly discriminable to older infants, was tested in Experiment 2, using a non-native Tigrinya distinction involving lips versus tongue tip. Both ages discriminated this between-organ contrast well, further supporting the AO hypothesis. Implications for theoretical accounts of infant speech perception are discussed. PMID- 14748446 TI - Early word learners' ability to access phonetic detail in well-known words. AB - Several recent studies from our laboratory have shown that 14-month-old infants have difficulty learning to associate two phonetically similar new words to two different objects when tested in the Switch task. Because the infants can discriminate the same phonetic detail that they fail to use in the associative word-learning situation, we have argued that this word-learning failure results from a processing overload. Here we explore how infants perform in the Switch task with already known minimally different words. The experiment involved the same phonetic difference as used in our earlier word-learning studies. Following habituation to two familiar minimal pair object-label combinations (ball and doll), infants of 14 months looked longer to a violation in the object-label pairing (e.g., label 'ball' paired with object doll) than to an appropriate pairing. These results using well known words are consistent with the pattern of data recently obtained by Swingley and Aslin (2002) in which it was found that infants of 14 months look longer to the correct object when the accompanying well known word is spoken correctly rather than mispronounced. We discuss how these results are compatible with the limited resource explanation originally offered by Stager and Werker (1997). PMID- 14748447 TI - Phonetic detail in the developing lexicon. AB - Although infants show remarkable sensitivity to linguistically relevant phonetic variation in speech, young children sometimes appear not to make use of this sensitivity. Here, children's knowledge of the sound-forms of familiar words was assessed using a visual fixation task. Dutch 19-month-olds were shown pairs of pictures and heard correct pronunciations and mispronunciations of familiar words naming one of the pictures. Mispronunciations were word-initial in Experiment 1 and word-medial in Experiment 2, and in both experiments involved substituting one segment with [d] (a common sound in Dutch) or [g] (a rare sound). In both experiments, word recognition performance was better for correct pronunciations than for mispronunciations involving either substituted consonant. These effects did not depend upon children's knowledge of lexical or nonlexical phonological neighbors of the tested words. The results indicate the encoding of phonetic detail in words at 19 months. PMID- 14748448 TI - Learning foreign vowels. AB - Two hypotheses have recently been put forward to account for listeners' ability to distinguish and learn contrasts between speech sounds in foreign languages. First, Best's Perceptual Assimilation Model and Flege's Speech Learning Model both predict that the ease with which a listener can tell one non-native phoneme from another varies directly with the extent to which these sounds assimilate to different native phonemes (Best, 1994; also Best, McRoberts, & Goodell, 2001; Flege, 1991). Second, Logan, Lively, & Pisoni (1991) have argued that training listeners to identify non-native phonemes teaches them sets of exemplars rather than more abstract distinctive feature values. I report here the results of three sets of experiments designed to test these hypotheses, in which American English listeners were trained to categorize German nonlow vowels. The first set of experiments show that some instances of the same contrast between German vowels are more easily discriminated than others, a result incompatible with the predictions of either Best's or Flege's models, but compatible with the alternative category recognition interpretation. The second set of experiments reveals effects of contextual and speaker variation on listeners' ability to learn [tense] but not [high] contrasts between foreign vowels, and are thus at least partly compatible with an exemplar model of foreign category learning (Pisoni, Lively, & Logan, 1994; also Nosofsky, 1986). The third set of experiments compares the predictions of Nosofsky's (1986) selective attention exemplar model of category learning with those of a feature learning model in tests of listeners' learning the natural classes to which the German vowels belong. The results are mixed: listeners learned the features that define the natural classes of [+/- high] and [+/- back] vowels, but could have learned either the feature that defines the natural classes of [+/- tense] vowels or sets of [+/- tense] exemplars. Natural classes defined by abstract distinctive feature values are thus learnable, even if their membership is phonetically polymorphous. PMID- 14748449 TI - Cracks in the system: professional and continuing education under scrutiny. PMID- 14748450 TI - Prevalence of psychopathology in preschool-age children. AB - PROBLEM: Little is known about psychopathology in preschool-age children. METHODS: A review of representative studies of psychiatric disorders in preschoolers. FINDINGS: Sample sizes ranged from 104 to 3,860 subjects, ages 1 to 9 years. Prevalence rates of psychiatric disorders varied from 0.1% to 26.4%; high rates of co-morbidity were reported. CONCLUSIONS: Studies addressing psychiatric disorders in preschoolers are extremely limited. Future research is needed to improve the diagnosis, treatment, and outcomes in preschool-age children. PMID- 14748451 TI - Cries from the morgue. AB - TOPIC: Family member homicide. PURPOSE: To explore the unique facets of bereavement for a family member of a homicide victim in order to assist clinicians during assessment, evaluation, and intervention with children. SOURCES: The authors' experience in working with children at a child grief center and published literature. CONCLUSIONS: Children experience grief uniquely and use differing methods of coping and adaptation. Knowledge of these differences can assist clinicians with promotion of adaptive coping and grieving in children who have been exposed to murder. PMID- 14748453 TI - Emphasizing quality in qualitative papers. PMID- 14748452 TI - Effects of communication skills training on parents and young adolescents from extreme family types. AB - PROBLEM: What were the effects of a communication skills training intervention among a sample of young adolescents and parents who scored in the "extreme" range of the Circumplex Model of Family Systems? METHODS: Thirty-seven young adolescents and a parent (intervention group) participated in communication skills training 2 hours/week for 6 weeks. Their responses on measures of satisfaction with the family system and perceptions of communication were compared with those of 47 young adolescents and a parent who scored in the extreme range but did not participate in the training (control group). FINDINGS: Fathers and young adolescents demonstrated no change as a result of the program. Mothers who participated in skills training perceived communication with their young adolescent as more open than control mothers, but became increasingly dissatisfied with the family system. CONCLUSION: This universal, community-based, family-focused intervention may not be indicated for extreme families. PMID- 14748454 TI - The physiology of deep-water running. AB - Deep-water running is performed in the deep end of a swimming pool, normally with the aid of a flotation vest. The method is used for purposes of preventing injury and promoting recovery from strenuous exercise and as a form of supplementary training for cardiovascular fitness. Both stroke volume and cardiac output increase during water immersion: an increase in blood volume largely offsets the cardiac decelerating reflex at rest. At submaximal exercise intensities, blood lactate responses to exercise during deep-water running are elevated in comparison to treadmill running at a given oxygen uptake (VO2). While VO2, minute ventilation and heart rate are decreased under maximal exercise conditions in the water, deep-water running nevertheless can be justified as providing an adequate stimulus for cardiovascular training. Responses to training programmes have confirmed the efficacy of deep-water running, although positive responses are most evident when measured in a water-based test. Aerobic performance is maintained with deep-water running for up to 6 weeks in trained endurance athletes; sedentary individuals benefit more than athletes in improving maximal oxygen uptake. There is some limited evidence of improvement in anaerobic measures and in upper body strength in individuals engaging in deep-water running. A reduction in spinal loading constitutes a role for deep-water running in the prevention of injury, while an alleviation of muscle soreness confirms its value in recovery training. Further research into the applications of deep-water running to exercise therapy and athletes' training is recommended. PMID- 14748455 TI - Motivation for physical activity in young people: entity and incremental beliefs about athletic ability. AB - Three studies are reported of children and youth aged 11-19 years (n = 3478) examining the nature of beliefs about athletic ability. Drawing on related research in academic, moral and stereotyping domains, development of a psychometric instrument assessing athletic ability beliefs is detailed. Support was found for a multidimensional hierarchical structure that is invariant across age and gender. Confirmatory factor analysis revealed a structure comprising two higher-order factors of entity and incremental beliefs underpinned by beliefs that athletic ability is stable and a gift (entity), and is open to improvement and can be developed through learning (incremental). Incremental beliefs, indirectly through a task goal orientation, and entity beliefs directly, predicted self-reported amotivation towards physical education and sport. On the other hand, enjoyment of physical activity in youth was predicted directly by task orientation and incremental beliefs. Predictions concerning the moderating role of perceived competence were not supported. Our findings highlight the importance of ability beliefs and goals in understanding the determinants of physical activity in children and youth. PMID- 14748456 TI - The effect of exercise training on the kinetics of the antibody response to influenza vaccination. AB - The significance of in vitro changes in immune function accompanying exercise training is unclear. To determine the effect of exercise on the response of the intact immune system to a challenge in vivo, we measured the speed and overall immunoglobulin G (IgG) response to influenza vaccination in humans engaged in different intensities of activity. Male participants (n = 21) were split into heavy and light training groups. Venous blood samples were collected 0, 2, 4, 7, 10 and 14 days after vaccination with trivalent influenza vaccine, and also 12 months after initial vaccination. Serum IgG was determined by enzyme-linked immunosorbant assay. There was a significant difference in baseline IgG between groups, but no difference in IgG concentration 14 days after vaccination. The IgG concentration remained elevated 12 months post-vaccination in the heavy training group. The results suggest a positive relationship between habitual physical activity and baseline antibody concentrations, which, in turn, affects the relative magnitude (fold or percentage increase) of the antibody response to vaccination. The training loads of the participants in this study had no effect on overall IgG measured 14 days after vaccination. PMID- 14748457 TI - Acute impact of submaximal resistance exercise on immunological and hormonal parameters in young men. AB - In this study, we examined the acute effects of submaximal resistance exercise on immunological and hormonal parameters in 7 resistance-trained and 10 non resistance-trained males. The participants, who were aged 29.5 +/- 7.1 years (mean +/- s), performed submaximal resistance exercise at 75% of their one repetition maximum. Blood samples were taken before, during, immediately after, and 30, 60 and 120 min after exercise and analysed for leukocyte subpopulations and stress hormones. Total leukocytes, neutrophils and monocytes increased during exercise, reaching their maximum 2 h after exercise. Lymphocytes increased during exercise, T-helper cells returned to resting values after exercise, and natural killer cells and T-suppressor cells decreased below resting values. The CD4/CD8 ratio decreased during exercise but increased during recovery. The resistance trained participants tended to have lower T-helper cell counts before, during and immediately after exercise and a lower CD4/CD8 ratio during recovery than the non resistance-trained participants. Plasma cortisol correlated positively with leukocytes during exercise (r = 0.572, P < 0.05), but negatively with T-helper cells 30 and 60 min after exercise (r = -0.573, P < 0.05; r = -0.642, P < 0.01, respectively). Our results indicate that resistance exercise leads to acute changes in leukocyte counts, despite moderate hormonal changes, independent of training status. Regular resistance exercise might lead to decreased T-helper cell counts and a lower CD4/CD8 ratio, which could increase susceptibility to infections. PMID- 14748458 TI - The effect of ball carrying method on sprint speed in rugby union football players. AB - Different methods of ball carrying can be used when a player runs with the ball in rugby union. We examined how three methods of ball carrying influenced sprinting speed: using both hands, under the left arm and under the right arm. These methods were compared with running without the ball. Our aim was to determine which method of ball carrying optimizes sprinting speed. Altogether, 48 rugby union players (age 21 +/- 2 years, height 1.83 +/- 0.1 m, body mass 85.3 +/ 12 kg, body fat 14 +/- 5%; mean +/- s) were recruited. The players performed twelve 30-m sprints in total (each player performed three trials under each of three methods of carrying the ball and sprinting without the ball). The design of the study was a form of Latin rectangle, balanced across the trial order for each of the methods and for pairwise combinations of the methods in blocks of four per trial. Each sprint consisted of a 10-m rolling start, followed by a 20-m timed section using electronic timing gates. Compared with sprinting 20 m without the ball (2.58 +/- 0.16 s), using both hands (2.62 +/- 0.16 s) led to a significantly slower time (P < 0.05). Sprinting 20 m with the ball under the left arm (2.61 +/- 0.15 s) or under the right arm (2.60 +/- 0.17 s) was significantly quicker than when using 'both hands' (P < 0.05), and both these methods were significantly slower than when running without the ball (P < 0.05). Accordingly, running with the ball in both hands led to the greatest decrement in sprinting performance, although carrying the ball under one arm also reduced the players' sprinting ability. Our results indicate that to gain a speed advantage players should carry the ball under one arm. PMID- 14748459 TI - The effect of pre-exercise carbohydrate feedings on the intensity that elicits maximal fat oxidation. AB - The aim of the present study was to examine the effect of ingesting 75 g of glucose 45 min before the start of a graded exercise test to exhaustion on the determination of the intensity that elicits maximal fat oxidation (Fatmax). Eleven moderately trained individuals (VO2max: 58.9 +/- 1.0 ml x kg(-1) x min( 1); mean +/- sx), who had fasted overnight, performed two graded exercise tests to exhaustion, one 45 min after ingesting a placebo drink and one 45 min after ingesting 75 g of carbohydrate in the form of glucose. The tests started at 95 W and the workload was increased by 35 W every 3 min. Gas exchange measures and heart rate were recorded throughout exercise. Fat oxidation rates were calculated using stoichiometric equations. Blood samples were collected at rest and at the end of each stage of the test. Maximal fat oxidation rates decreased from 0.46 +/ 0.06 to 0.33 +/- 0.06 g min(-1) when carbohydrate was ingested before the start of exercise (P < 0.01). There was also a decrease in the intensity which elicited maximal fat oxidation (60.1 +/- 1.9% vs 52.0+3.4% VO2max) after carbohydrate ingestion (P < 0.05). Maximal power output was higher in the carbohydrate than in the placebo trial (346 +/- 12 vs 332 +/- 12 W) (P < 0.05). In conclusion, the ingestion of 75 g of carbohydrate 45 min before the onset of exercise decreased Fatmax by 14%, while the maximal rate of fat oxidation decreased by 28%. PMID- 14748460 TI - The effect of wet suit use by triathletes: an analysis of the different phases of arm movement. AB - We analysed stroke phases and arm and leg coordination during front crawl swimming with and without a wet suit. Twelve nationally and internationally ranked French male triathletes performed three swim trials in randomized order using the front crawl stroke with and without a wet suit. All triathletes swam at three different swim velocities, corresponding to the paces appropriate for the 800 m (V800), 100 m (V100) and 50 m (V50) events. The different stroke phases and arm and leg coordination were identified by video analysis. Arm coordination was quantified using a new index of coordination, which expresses the three major modalities of opposition, catch-up and superposition in swimming. At all swim velocities, no significant differences in leg movements with or without the wet suit were noted. However, the wearing of the wet suit was associated with a significantly greater stroke length at the paces appropriate for the 100 and 50 m events (+3.46% and +3.10% at V100 and V50, respectively; P<0.01); a significantly greater stroke index at all three velocities (+5.18%, +5.21% and +5.91% at V800, V100 and V50, respectively; P<0.01); a significantly shorter pulling phase ( 10.97%; P<0.05) and lower index of coordination (-21.87%; P<0.01) at the pace appropriate for the 800 m; and a significantly greater entry and catch phase (+9.81%; P<0.05) at the pace appropriate for the 100 m. We conclude that the wet suit amplified the coordination mode of the triathletes (i.e. catch-up coordination) without modifying stroke rate, recovery phase or leg movements. PMID- 14748461 TI - Shiga toxin producing Escherichia coli infection: current progress & future challenges. AB - Shiga toxin producing Escherichia coli (STEC) is a newly emerged pathogen that has been the focus of immense international research effort driven by its recognition as a major cause of large scale epidemics and thousands of sporadic cases of gastrointestinal illness. It produces a severe bloody diarrhoea that is clinically distinct from other types of diarrhoeal diseases caused by other enteric pathogens. One of the most important areas of current exploration concerns how STEC enters our food chain, an investigational avenue that begins with the ecology of STEC in animals and in the environment. A variety of foods have been identified as vehicles of STEC-associated illness and this makes the organism one of the most serious threats to the food industry in recent years. The pathogenesis of STEC is multifactorial and involves several levels of interaction between the bacterium and the host. STEC strains carry a set of virulence genes that encode the factors for attachment to host cells, elaboration of effective molecules and production of two different types of Shiga toxins. These genes are found in the locus of enterocyte effacement (LEE), lamboid phages, and a large virulence associated plasmid. The publication of the complete genome sequence of Esch. coli O157:H7 chromosome offers a unique resource that will help to identify additional virulence genes, to develop better methods of strain detection and in the understanding of the evolution of Esch. coli through comparison with the genome of the non-pathogenic laboratory strain Esch. coli K 12. These research efforts in turn, should lead to development of new potent and cost effective anti-Stx therapies or vaccines and thereby major improvement in human health world-wide. PMID- 14748462 TI - Antimicrobial resistance in Enterococcus faecalis at a tertiary care centre of northern India. AB - BACKGROUND & OBJECTIVES: Multiresistant enterococci are emerging as a leading nosocomial pathogen. Knowledge of the profile of antimicrobial resistance is essential to formulate treatment guidelines for infections caused by enterococci. This study reports the antimicrobial sensitivity of enterococci isolated during a one year period from clinical samples of patients admitted to a teriary care hospital of Delhi. METHODS: A total of 444 isolates of Enterococcus faecalis were screened for antimicrobial susceptibility by the disk diffusion technique as recommended by the National Committee for Clinical Laboratory Standards (NCCLS). Screening for vancomycin resistance was done by the vancomycin screen agar method recommended by NCCLS, which was confirmed by determination of minimum inhibitory concentration (MIC) using microbroth dilution and E-test methods. Vancomycin resistance phenotypes were determined by polymerase chain reaction. RESULTS: A total of 115 (26%) isolates had high level aminoglycoside resistance, 293 (66%) were resistant to ampicillin, 391 (88%) to ciprofloxacin and 377 (85%) to erythromycin. Vancomycin resistance was found in five (1%) isolates, of which four had van A phenotype and one had van B phenotype. INTERPRETATION & CONCLUSION: Emergence of vancomycin resistant enterococci is of concern due to the limited therapeutic options. Implementation of infection control measures can contain the spread of these resistant bacteria. PMID- 14748463 TI - Occurrence & detection of AmpC beta-lactamases at a referral hospital in Karnataka. AB - BACKGROUND & OBJECTIVES: AmpC beta-lactamases confer resistance to a wide variety of beta-lactam drugs except for cefepime, cefpirome and carbapenems. They are known to be responsible for nosocomial outbreaks, therapeutic failures and multidrug resistance. Although reported with increasing frequency the true rate of occurrence of these beta-lactamases in Enterobacteriaceae is not known. Hence the present study was undertaken to determine the occurrence of AmpC enzymes among clinical isolates. METHODS: A total of 520 consecutive, non-repeat clinical isolates were included in the present study. Twenty eight strains resistant to cefoxitin were tested for AmpC beta-lactamases by the modified 3-dimensional extract method. Isolates harbouring AmpC beta-lactamases were tested for inducible beta-lactamases by disc diffusion. RESULTS: Sixteen (3.3%) isolates were positive for AmpC beta-lactamases. Based on the species 9 (3.3%) Escherichia coli, 4 (2.2%) Klebsiella pneumoniae, 2 (5%) Citrobacter freundii and 1 (5.5%) isolate of Enterobacter aerogenes harboured AmpC enzymes. Nine (56.3%) of AmpC harbouring strains, were urinary isolates. All the isolates were sensitive to imipenem and variably sensitive to aminoglycosides and co-trimoxazole. INTERPRETATION & CONCLUSION: Our findings document the presence of AmpC enzymes in this region. Hence AmpC beta-lactamase detection should be undertaken in clinical isolates showing resistance to broad-spectrum cephalosporins. PMID- 14748464 TI - Comparison of ondansetron with metoclopramide in prevention of acute emesis associated with low dose & high dose cisplatin chemotherapy. AB - BACKGROUND & OBJECTIVES: Nausea and vomiting remain the most distressing side effects of cancer chemotherapy. The present study aimed to study the efficacy and tolerability of ondansetron versus (vs) metoclopramide in different dose related grades of cisplatin induced acute emesis. METHODS: A total of 137 patients were enrolled and 80 completed the study. Cisplatin 60 mg/m2 was given intravenously (iv) either as a single dose on day 1 (high dose regimen) or in three doses of 20 mg/m2 each on days 1-3 (low dose regimen) along with bleomycin +5-flurouracil in 40 patients each. Patients were randomized in each cisplatin regimen to receive either 20 mg metoclopramide (20 patients) or 8 mg ondansetron (20 patients) iv 30 min prior to cisplatin administration followed by 20 mg metoclopramide or 8 mg ondansetron orally 8 h respectively for 24 h after the last cisplatin administration. Ten patients receiving high dose cisplatin in each group were also given dexamethasone 8 mg iv with the primary antiemetic. Patients were assessed for 24 h after the last cisplatin injection. RESULTS: In low dose cisplatin regimen, complete suppression of acute emesis occurred in 65 per cent patients receiving ondansetron versus 30 per cent receiving metoclopramide, while in high dose regimen, complete response rate was 20 per cent with ondansetron versus 0 per cent with metoclopramide. Dexamethasone significantly augmented the antiemetic efficacy of metoclopramide but not that of ondansetron. Protection from nausea in the acute phase was seen in 95 per cent patients receiving ondansetron vs 70 per cent receiving metoclopramide in low dose regimen. With high dose the protection rates were 90 vs 0 per cent respectively. Combination of dexamethasone + metoclopramide achieved 70 per cent protection while dexamethasone + ondansetron was effective in 90 per cent. Dropouts and withdrawals were more among patients receiving high dose cisplatin and antiemetic regimens without dexamethasone. Thirty nine adverse events were reported by 20 out of 80 patients. All adverse events were mild. INTERPRETATION & CONCLUSION: The results demonstrate dose related emetogenicity of cisplatin and superior antiemetic efficacy of ondansetron, especially against high dose cisplatin regimen. Dexamethasone potentiated efficacy of metoclopramide but not that of ondansetron. The combination of metoclopramide plus dexamethasone was found to be as efficacious as ondansetron monotherapy. PMID- 14748465 TI - Effect of sodium citrate ingestion on oxygen debt & exercise endurance during supramaximal exercise. AB - BACKGROUND & OBJECTIVES: Administration of alkalinizing agents has been shown to improve performance in high intensity exercise. The present investigation was undertaken to determine whether sodium citrate ingestion enhances supramaximal endurance performance on a cycle ergometer and to assess its effect on oxygen debt incurred for the same supramaximal energy output. METHODS: Ten untrained, healthy, males performed acute intense bicycle ergometry exercise in two sessions viz., without (control) and with sodium citrate ingestion (experimental) in a dose of 0.5 g/kg body weight. Pre-exercise O2 consumption, pulse rate (PR) and respiratory rate (RR)/minute were recorded before both sessions. Exhaustion time (ET) was noted and work done calculated in the control session. Post-exercise PR and RR/min were also recorded in both sessions of study. Venous blood samples were drawn post-exercise and analysed for pH and lactate. O2 debt incurred was calculated in both sessions. RESULTS: Work output in supramaximal exercise averaged 69.40 +/- 15.31 Watts in the control session. None of the subjects in the experimental session complained of fatigue even when the exhaustion time noted in the control session was reached. Post-exercise tachycardia and tachypnoea were both significantly less (P<0.05 and P<0.001 respectively) in subjects after the experimental session as compared to the control. O2 debt incurred and lowering of blood pH were also significantly less (P<0.001) in the post-citrate phase. However, serum lactate increased significantly (P<0.05) in subjects after the experimental session. INTERPRETATION & CONCLUSION: Ingestion of sodium citrate prior to supramaximal exercise resulted in a reduction in post exercise O2 debt incurred. Blood pH was almost normal despite a higher serum lactate concentration. This probably led to an enhancement in exercise performance. PMID- 14748466 TI - Induction & resolution of lobar pneumonia following intranasal instillation with Klebsiella pneumoniae in mice. AB - BACKGROUND & OBJECTIVES: Pneumonia caused by Klebsiella pneumoniae is important due to its high morbidity and mortality, especially in context of nosocomial infections. Many experimental studies have focused on the induction and progression of infection till it peaks, but the process of resolution has not been described. In the present study, we successfully attempted to establish an acute respiratory tract infection model in BALB/c strain of mice with K. pneumoniae employing a simple, reproducible intranasal instillation method. METHODS: Experimental pneumonia was induced by two strains of K. pneumoniae in BALB/c mice following intranasal instillation, and the course of pneumonia was studied by bacteriological and histopathological evaluation of the lung tissue. RESULTS: Both the strains were similar in their ability to induce infection which peaked on day 3, post infection. However, a strain dependent difference in relation to bacterial load and the process of resolution was observed. INTERPRETATION & CONCLUSION: The present study provides a model of lobar pneumonia produced by K. pneumoniae which can be useful for studying therapeutic and preventive interventions. PMID- 14748467 TI - The genetics of diabetes mellitus. AB - Genes play an important role in the development of diabetes mellitus. Putative susceptibility genes could be the key to the development of diabetes. Type 1 diabetes mellitus is one of the most common chronic diseases of childhood. A combination of genetic and environmental factors is most likely the cause of Type 1 diabetes. The pathogenetic sequence leading to the selective autoimmune destruction of islet beta-cells and development of Type 1 diabetes involves genetic factors, environmental factors, immune regulation and chemical mediators. Unlike Type 1 diabetes mellitus, Type 2 diabetes is often considered a polygenic disorder with multiple genes located on different chromosomes being associated with this condition. This is further complicated by numerous environmental factors which also contribute to the clinical manifestation of the disorder in genetically predisposed persons. Only a minority of cases of type 2 diabetes are caused by single gene defects such as maturity onset diabetes of the young (MODY), syndrome of insulin resistance (insulin receptor defect) and maternally inherited diabetes and deafness (mitochondrial gene defect). Although Type 2 diabetes mellitus appears in almost epidemic proportions our knowledge of the mechanism of this disease is limited. More information about insulin secretion and action and the genetic variability of the various factors involved will contribute to better understanding and classification of this group of diseases. This article discusses the results of various genetic studies on diabetes with special reference to Indian population. PMID- 14748468 TI - Increasing HIV seropositivity among adult tuberculosis patients in Delhi. AB - We report the HIV seropositivity among adult TB patients from our hospital, a tertiary care hospital in north India between 2000-2002. Of the 555 patients with various forms of tuberculosis, 52 were found to be seropositive (9.4%). In 1994 1999, the HIV seropositivity in this hospital was only 0.4 per cent (2 of 500 patients). This communication describes a dramatic increase in seropositivity and highlights the importance of continued HIV serosurveillance in patients with TB. PMID- 14748469 TI - A recent outbreak of cholera due to Vibrio cholerae O1 Ogawa in & around Chandigarh, North India. AB - An outbreak of cholera caused by Vibrio cholerae O1 Ogawa occurred in and around Chandigarh during July 22-31, 2002. Of the 303 patients admitted to two hospitals, 82 were confirmed by culture. Two rehabilitation colonies located at the periphery of Chandigarh were mainly affected. The isolates were biotyped as Eltor and were susceptible to many antibiotics. Thirty one (35.2%) of 88 water samples showed evidence of faecal contamination. The survey of the area revealed sewage contamination of the drinking water supply. The outbreak was controlled by providing safe drinking water to the people and correcting the defects in the sewage and water pipelines. PMID- 14748470 TI - Rapid diagnosis of vaginal carriage of group B beta haemolytic streptococcus by an enrichment cum antigen detection test. AB - BACKGROUND & OBJECTIVES: Group B beta haemolytic streptococcus (GBS) is a frequent colonizer of the maternal genital tract causing peripartum fever, puerperal sepsis, neonatal sepsis and neonatal meningitis. The conventional methods for detection of maternal colonization take 24-48 h. We made an attempt to standardize a rapid enrichment cum antigen detection test to screen pregnant women for GBS colonization in less than 8 h, so as to enable early institution of measures to prevent neonatal sepsis. METHODS: Vaginal swabs of 100 women >36 wk of gestation were inoculated onto enrichment broth (Todd Hewitt broth with lysed horse blood and antibiotics). After incubation for 1,2,4,6, and 18 h, the broth was cultured on sheep blood agar. In culture positive cases, the enrichment broth was subjected to antigen detection by latex agglutination test (LAT). For further evaluation of the rapid test, another group of 100 pregnant women were screened for GBS carriage by 6 h enrichment broth culture followed by antigen detection test. RESULTS: Five of the first group yielded GBS on culture and all were positive for GBS antigen after 6 h enrichment. Thirteen of the second group were positive for the antigen, but GBS could be isolated in ten only. This enrichment cum antigen detection test showed sensitivity, specificity, and positive and negative predictive values of 100, 98.4, 83.3 and 100 per cent respectively and could detect as few as 10(3) cfu/ml organisms. Maternal vaginal carriage of GBS was 7.5 per cent (15/200). INTERPRETATION & CONCLUSION: Six hours of enrichment followed by antigen detection proved to be a rapid and reliable method for detection of GBS colonization. This test is easy to perform making it an ideal test for screening GBS vaginal colonization at labour and starting chemoprophylaxis, where indicated on the same day, before the woman is discharged. PMID- 14748472 TI - Hand anthropometry of Indian women. AB - BACKGROUND & OBJECTIVES: Data on the physical dimension of the hand of Indian women are scanty. This information is necessary to ascertain human-machine compatibility in the design of manual systems for the bare and gloved hand, such as design and sizing of hand tools, controls, knobs and other applications in different kinds of precision and power grips. The present study was undertaken to generate hand anthropometric data of 95 women, working in informal industries (beedi, agarbatti and garment making). METHODS: Fifty one hand measurements of the right hand (lengths, breadths, circumferences, depths, spreads and clearances of hand and fingers) were taken, using anthropometric sliding and spreading calipers, measuring tape and handgrip strength dynamometer. The data were statistically analyzed to determine the normality of data and the percentile values of different hand dimensions, and simple and multiple regression analysis were done to determine better predictors of hand length and grip strength. RESULTS: The hand breadths, circumferences and depths were approximately normally distributed, with some deviation in case of the finger lengths. Hand length was significantly correlated with the fist, wrist and finger circumferences. The fist and wrist circumferences, in combination, were better predictors of hand length. The hand lengths, breadths and depths, including finger joints of the Indian women studied were smaller than those of American, British and West Indian women. The hand circumferences of the Indian women were also smaller than the American women. Grip strengths of Indian women (20.36 +/- 3.24 kg) were less than those of American, British and West Indian women. Grip strength was found to be statistically significant with hand dimensions, such as hand height perpendicular to wrist crease (digit 5), proximal interphalangeal joint breadth (digit 3) and hand spread across wedge 1. INTERPRETATION & CONCLUSION: The women who are forced to frequently use cutters, strippers and other tools, which are not optimally designed to their hand dimensions and strength range, might have higher prevalence of clinical symptoms and disorders of the hand. In view of the human hand-tool interface requirements, the present data on Indian women would be useful for ergo-design applications of hand tools and devices. PMID- 14748471 TI - Oxidative damage in intermediate syndrome of acute organophosphorous poisoning. AB - BACKGROUND & OBJECTIVES: The main cause of morbidity due to organophosphate poisoning is intermediate syndrome (Type II paralysis) that can occur 48-72 h after poisoning. Mechanisms that underlie the intermediate syndrome are not known. This study investigates the role of oxidative damage to muscles as a possible mechanism underlying the development of the intermediate syndrome. METHODS: Nineteen patients with acute organophosphate poisoning were evaluated from admission to discharge from intensive care for the severity of poisoning and the development and duration of the intermediate syndrome. Blood cholinesterases and parameters of oxidative stress were studied daily and their temporal profiles analysed according to the severity of poisoning and the development and duration of the intermediate syndrome. RESULTS: Fifteen patients had severe poisoning and 16 developed intermediate syndrome. There was a positive association between the severity of poisoning and the occurrence of intermediate syndrome. There was no association between the organophosphate ingested and the development of intermediate syndrome. Erythrocyte membrane acetylcholinesterase and serum butyrylcholinesterase levels at admission and over the course of poisoning were significantly (P < 0.001) reduced in patients compared to controls. There were significantly (P < 0.05) higher levels of lipid peroxidation, conjugated dienes and protein thiols in erythrocyte membranes of patients who developed the intermediate syndrome compared to healthy controls, in patients who developed intermediate syndrome compared to those who did not and in patients with long compared to short duration intermediate syndrome. INTERPRETATION & CONCLUSION: In acute organophosphate poisoning, severe and prolonged acetylcholinesterase inhibition is associated with oxidative stress, detected in erythrocyte membranes, that occurs early in the course of poisoning and may contribute to the development and severity of intermediate syndrome. PMID- 14748473 TI - Physical fitness in persons with hemiparetic stroke. AB - In persons with hemiparetic stroke, assessment and promotion of fitness have so far received limited attention, partly because of the lack of appropriate measures applicable to them. Because these mobility-impaired persons are prone to inactivity, disuse and insulin resistance are likely to occur, and can aggravate the already significant health and economic consequences that stroke entails. It is therefore important to assess objectively their fitness to devize effective and efficient fitness promotion programs. Because of physical limitations, however, many persons with stroke cannot perform traditional stress testing using a treadmill or a cycle ergometer, and maximal oxygen consumption, which is regarded as a gold standard, is not a practical measure. In this article, we reviewed the current status of research on fitness in persons with hemiparetic stroke from the perspectives of evaluation, structure analysis of fitness, and longitudinal changes during a rehabilitation program. As a measure of fitness, indices obtainable with a submaximal exercise are proposed, such as anaerobic threshold and heart rate oxygen coefficient. Protocols applicable to persons with hemiparetic stroke with a variety of functional limitations have been developed (basic bedside activities, bridging activity, or single arm ergometry). The structure of their fitness is demonstrated to be described by a fitness model of healthy persons (cardiopulmonary, muscular and metabolic dimensions) if the paresis/activities of daily living dimension is added. Several studies suggest that fitness improves during a conventional stroke rehabilitation program. Studying the changes of the above four dimensions can help develop more effective fitness training programs. PMID- 14748474 TI - HLA class I defects in malignant lesions: what have we learned? AB - Depending on the tumor types, HLA class I antigen downregulation or loss has been found in 16% to 50% of malignant lesions in many malignancies with a clinical association with histopathological markers of poor prognosis of the disease and with reduced free interval and survival. These findings may reflect the escape of tumor cells with HLA class I abnormalities from recognition and destruction by HLA class I-restricted, tumor-associated antigen-specific cytotoxic T lymphocytes. This possibility has stimulated investigations on the mechanisms underlying HLA class I antigen abnormalities in malignant cells. Distinct molecular defects underlying an abnormal HLA class I phenotype have been identified and characterized. These defects range from structural alterations of the genes which encode HLA class I antigen subunits to deregulation of antigen processing machinery components responsible for a functional HLA class I antigen expression. These findings, in conjunction with those of clinical recurrence of lesions with HLA class I antigen loss following T cell-based immunotherapy in patients, suggests that immunoselection may play a role in the generation of malignant lesions with HLA class I antigen abnormalities. This possibility has stressed the need to effectively monitor functional HLA class I antigen expression in malignant lesions in the application of T cell-based immunotherapy as well as to develop strategies to circumvent the negative impact of immunoselection. PMID- 14748475 TI - Comparative effects of treatment with etidronate and alendronate on bone resorption, back pain, and activities of daily living in elderly women with vertebral fractures. AB - The purpose of the present study was to compare the effects of treatment with etidronate and alendronate on bone resorption, back pain, and activities of daily living (ADL) in elderly women with vertebral fractures. Fifty elderly women, 63 84 years of age, with back pain due to osteoporotic vertebral fractures were randomly divided into two groups with 25 patients in each group: the cyclical etidronate treatment group (200 mg/day for 2 weeks per 3 months) and the alendronate treatment group (5 mg/day). The level of urinary cross-linked N terminal telopeptides of type I collagen (NTx) measured by an enzyme-linked immunosorbent assay, back pain evaluated with the face scale score, and the ADL score (disability) determined with a questionnaire were assessed before and 3 and 6 months after the start of treatment. No significant differences in these parameters were found between the two groups before the treatment. The urinary NTx level, the face scale score, and the ADL score decreased significantly in both groups. Although the reduction in the urinary NTx level was significantly greater in the alendronate group than in the etidronate group, the reduction in the face scale score was transiently significantly greater in the etidronate group than in the alendronate group. However, changes in the ADL score did not significantly differ between the two groups. The present study showed that although back pain was reduced and ADL was improved in both treatment groups of elderly women with vertebral fractures, the mechanism for the reduction in back pain differs to some extent between the two treatment groups. A double-blind placebo-controlled study is needed to confirm the therapeutic effects of these agents on back pain and deterioration of ADL. PMID- 14748476 TI - Roots of Empathy: responsive parenting, caring societies. AB - What is common in aggression and in abusive/neglectful parenting is low levels of empathy. Fostering empathy--the ability to identify with another person's feelings--can serve as an antidote to aggression and is crucial to good parenting. Poor parenting and aggression cut across all socioeconomic levels of the community and, as such, empathy needs to be fostered in all children. During the period of rapid brain development, adversity has a devastating impact on the baby's developing brain. Repeated experiences of stress are hardwired into the brain, creating damaging pathways. Risk factors such as domestic violence, child abuse and neglect, maternal depression, maternal addictions, and poverty are not just additive to the vulnerable developing brain; they are multiplicative in their impact. The parent is the baby's lifeline, mitigating stress for them and helping them to learn to regulate their emotions. The impact of poor parenting on a child's life is profound, resulting in insecure attachments which lead to a spectrum of inadequate coping mechanisms, poor emotional regulation, diminished learning potential and low competence. Responsive and nurturing parenting is the key to optimal early childhood development; it allows the young brain to develop in a way that is less aggressive and more emotionally stable, social and empathic. Good early childhood development leads to good human development. We must match our investment where the opportunity is most ripe--building parenting capacity. The 'Roots of Empathy' program offers real hope in breaking the intergenerational transference of poor parenting and violence. PMID- 14748477 TI - Dynamics of calcium and its roles in the dendrite of the cerebellar Purkinje cell. AB - The calcium ion (Ca2+) serves as an important cellular messenger with spatio temporally highly dynamic patterns. Not only Ca2+ entering from the plasma membrane but also Ca2+ released from intracellular store sites play crucial roles in neurons as well as in other cell types. The cerebellar Purkinje cell shows a variety of spatio-temporal Ca2+ dynamics in its rich arborization, and the Ca2+ release mechanism mediated by IP3 receptors and ryanodine receptors has been shown to be crucial for the induction of long-term depression, a form of synaptic plasticity, in this neuron. Purkinje cells in a model mouse line of human neurodegenerative disease, spinocerebellar ataxia type 1, showed abnormal Ca2+ release properties, which indicated tight regulation of Ca2+ dynamics in the wild type Purkinje cell. Quantitative analysis of the complex Ca2+ dynamics in the Purkinje cell dendrite is still in progress to elucidate the link between external stimuli and the resultant biochemical changes. PMID- 14748478 TI - A 38-year-old man with pulmonary hypertension, who had undergone atrial septal closure 26 years previously. AB - The patient was a 38-year-old man. He underwent atrial septal closure at the age of 12 years at Yokohama City University Hospital, when he already had pulmonary vascular change and reduced left-to-right shunt with Qp/Qs of 1.55 and pulmonary artery pressure (PA) of 56/22 mmHg. Thereafter, he enjoyed running and skiing without any symptoms up until 32 years of age, when he developed syncope due to severe pulmonary hypertension and atrial flutter. PA was 116/57 mmHg and mRA was 13 mmHg on cardiac catheterization. He developed right heart failure and was referred to Keio University Hospital on May 12th, 2001. Home intravenous prostacyclin infusion therapy was introduced in addition to treatment for right heart failure. Echocardiography revealed a residual interatrial shunt (from right to left). He recovered and was discharged. His condition worsened again and he was readmitted to our hospital with chief complaint of visual disturbance due to digoxin intoxication, in addition to right heart failure. Despite aggressive treatment, he died of severe pulmonary hypertension, right heart failure and congestive hepatic failure on December 10th, 2001. The differential diagnosis, pathophysiology and necessary treatment of pulmonary hypertension are discussed in this paper. The clinical diagnosis was Eisenmenger syndrome due to atrial septal defect, and the pathological findings were compatible with this. PMID- 14748479 TI - The Keio University Medical Science Fund (KUMSF). The 2003 Keio Medical Science Prize awarded. PMID- 14748480 TI - ECG of the month. Rave. Gamma-hydroxybutyric acid intoxication. PMID- 14748481 TI - Radiology case of the month. A subtle cerebellar abnormality. Acute cerebellar ischemia (stroke). PMID- 14748482 TI - Pathology case of the month. A cheek mass in a young woman. Subcutaneous panniculitis-like T-cell lymphoma. PMID- 14748483 TI - Inverting papilloma of the nose and paranasal sinuses. AB - Inverting papilloma is a very uncommon neoplasm; however, its ability to destroy bone, recur after incomplete removal, and undergo malignant transformation makes it an important disease to recognize. Because early diagnosis and prompt treatment offer high rates of cure, it is important to be able to differentiate inverting papilloma from more benign nasal growths such as allergic nasal polyps. This article will summarize the etiology, location, signs, symptoms, potential for malignancy, and current treatment strategies for inverting papilloma. PMID- 14748484 TI - A 71-year-old man with hyperglycemia and mental status changes. AB - Hyperglycemic hyperosmolar syndrome is an extreme but relatively common presentation of uncontrolled or new-onset diabetes mellitus. The diagnosis of the disorder itself is fairly straightforward, but the search for an underlying cause can be challenging. Infections are the usual precipitating factor, but a variety of other stressors can be involved. We report herein a patient presenting with hyperglycemic hyperosmolar coma with three possible precipitating infections: pharyngitis, urinary tract infection, and infective endocarditis. PMID- 14748485 TI - Prevalence of the Brugada electrocardiographic pattern at the Medical Center of Louisiana in New Orleans. AB - Since 1992 the Brugada syndrome has gained recognition as a cause of ventricular fibrillation. The syndrome was originally described in patients with the diagnostic triad of (1) right bundle branch block, (2) an electrocardiogram (ECG) with persistent ST-segment elevation in leads V1, V2, and V3, and (3) sudden cardiac death. Two different types of ST-segment elevation, coved and saddleback, have been described. All patients originally described had structurally normal hearts. The definition of the Brugada electrocardiogram (originally right bundle branch block and ST-segment elevation in V1, V2, and V3 in characteristic coved or saddleback configuration) has been evolving since the initial description, and not all patients with the Brugada electrocardiogram have the Brugada syndrome. We designed a trial to determine the prevalence in our population at the Medical Center of Louisiana in New Orleans of the Brugada ECG as it was originally defined. ECGs performed in 1997 were examined for changes consistent with the Brugada electrocardiogram. Those ECGs with changes secondary to another identifiable cause were excluded. The amount and type of ST-segment elevation in leads V1, V2, and V3 were recorded for the remaining ECGs. From a total of 55,446 electrocardiograms performed on 27,328 patients, we were able to identify only 18 ECGs with the changes originally described by Brugada, and none of them meet current criteria. Our study suggests that in our patient population the ECG now considered typical of the Brugada syndrome is rare. PMID- 14748486 TI - Economic benefits of establishing a National Cancer Center in Louisiana. AB - This paper estimates the economic benefits of establishing a National Cancer Institute-designated center in Louisiana. Estimates of direct and indirect costs of cancer were used to derive the potential benefits of a cancer center. A fully functional cancer center should be able to prevent about 6,550 deaths over the next 10-year period implying a savings of about 23,000 disability-adjusted life years. The potential revenues to be generated through new patients from neighboring states, diversion of Louisiana patients back to the state from other out-of state facilities, and the indirect benefits derived through increased economic productivity (due to life years saved) makes the establishment of the center highly cost-effective. The benefit-cost ratio of establishing a cancer center becomes 8.5, i.e., spending one million dollars in a cancer center should generate about dollars 8.5 million worth of economic benefits for the State over a 10-year time horizon. PMID- 14748487 TI - Total lung preservation following mainstem-bronchial resection for carcinoid tumor. AB - Carcinoid tumors of the trachea and bronchi account for approximately 2% of all lung tumors. Carcinoid tumors, especially those in the bronchi, often produce obstruction that can mimic foreign body aspiration or asthma and may lead to post obstruction atelectasis and pneumonia. Management of carcinoid tumors of the bronchi has been by endoscopic resection, sleeve lobectomy with parenchymal sparing, and pneumonectomy. We present a case in which an obstructing bronchial carcinoid tumor was successfully managed by long-segment mainstem-bronchial resection with total lung preservation by re-implantation. PMID- 14748488 TI - Postprandial hyperglycemia. AB - Much attention has been paid to fasting glucose levels in both the diagnosis and management of diabetes mellitus. This test is well ingrained in clinical practice and well accepted by our patients. However, does a fasting glucose level give the clinician a complete picture? Is postprandial hyperglycemia associated with adverse events? This article will explore postprandial glucose levels and compare them to fasting glucose levels. The importance of postprandial and post-challenge (2 hours after a standard oral glucose tolerance test) glucose levels will be discussed. Drugs which address postprandial hyperglycemia will be reviewed briefly. PMID- 14748489 TI - Treatment of toxic epidermal necrolysis with intravenous immunoglobulin. AB - Toxic epidermal necrolysis is a severe adverse drug reaction that produces extensive mucocutaneous damage, with full-thickness epidermal detachment, and has many clinical similarities to severe burn injuries. The treatment is mainly supportive and aimed at preventing complications while the disease takes its natural course, and the skin reepithelializes. Much interest exists in the development of a specific therapy targeted at the disease process itself. Because the diagnosis has an incidence of only 0.5-1 case/million/year, large controlled studies are lacking, but a recent, better understanding of this disease has provided the rationale for the use of intravenous immunoglobulin. We present a case of toxic epidermal necrolysis that showed a good response to intravenous immunoglobulin G and review the recent literature condition and its management. PMID- 14748490 TI - The Medical Education Commission Report 2003: GME production renews physician supply. AB - Louisiana data for the match entry into Graduate Medical Education, and the renewal of the practicing physician workforce, are essentially parallel to comparable data for the United States as a whole. The State of Louisiana Medical Education Commission offers reports and publications in the Journal of the Louisiana State Medical Society compiling yearly and trend data and analysis. The 2003 result of the match follows the trend of successful completion over the last five years. While Louisiana is similar in many comparable categories to the United States, the state is somewhat higher in primary care growth, now stabilized and leveling, as are total GME and total physicians. Relatively small changes may be baseline fluctuations, and not a trend of longer-term change. The GME system is remarkably stable and successful overall, though complex right down to individual variation. PMID- 14748491 TI - Advice from the past hope for the future. PMID- 14748492 TI - QMLE: fast, robust, and efficient estimation of distribution functions based on quantiles. AB - Quantile maximum likelihood (QML) is an estimation technique, proposed by Heathcote, Brown, and Mewhort (2002), that provides robust and efficient estimates of distribution parameters, typically for response time data, in sample sizes as small as 40 observations. In view of the computational difficulty inherent in implementing QML, we provide open-source Fortran 90 code that calculates QML estimates for parameters of the ex-Gaussian distribution, as well as standard maximum likelihood estimates. We show that parameter estimates from QML are asymptotically unbiased and normally distributed. Our software provides asymptotically correct standard error and parameter intercorrelation estimates, as well as producing the outputs required for constructing quantile-quantile plots. The code is parallelizable and can easily be modified to estimate parameters from other distributions. Compiled binaries, as well as the source code, example analysis files, and a detailed manual, are available for free on the Internet. PMID- 14748493 TI - Testing curvatures of learning functions on individual trial and block average data. AB - Many models offer different explanations of learning processes, some of them predicting equal learning rates between conditions. The simplest method by which to assess this equality is to evaluate the curvature parameter for each condition, followed by a statistical test. However, this approach is highly dependent on the fitting procedure, which may come with built-in biases difficult to identify. Averaging the data per block of training would help reduce the noise present in the trial data, but averaging introduces a severe distortion on the curve, which can no longer be fitted by the original function. In this article, we first demonstrate what is the distortion resulting from block averaging. The block average learning function, once known, can be used to extract parameters when the performance is averaged over blocks or sessions. The use of averages eliminates an important part of the noise present in the data and allows good recovery of the learning curve parameters. Equality of curvatures can be tested with a test of linear hypothesis. This method can be performed on trial data or block average data, but it is more powerful with block average data. PMID- 14748494 TI - Estimating the mean effect size in meta-analysis: bias, precision, and mean squared error of different weighting methods. AB - Although use of the standardized mean difference in meta-analysis is appealing for several reasons, there are some drawbacks. In this article, we focus on the following problem: that a precision-weighted mean of the observed effect sizes results in a biased estimate of the mean standardized mean difference. This bias is due to the fact that the weight given to an observed effect size depends on this observed effect size. In order to eliminate the bias, Hedges and Olkin (1985) proposed using the mean effect size estimate to calculate the weights. In the article, we propose a third alternative for calculating the weights: using empirical Bayes estimates of the effect sizes. In a simulation study, these three approaches are compared. The mean squared error (MSE) is used as the criterion by which to evaluate the resulting estimates of the mean effect size. For a meta analytic dataset with a small number of studies, the MSE is usually smallest when the ordinary procedure is used, whereas for a moderate or large number of studies, the procedures yielding the best results are the empirical Bayes procedure and the procedure of Hedges and Olkin, respectively. PMID- 14748495 TI - On-line simulations of models for backward masking. AB - Five simulations of quantitative models of visual backward masking are available on the Internet at http://www.psych.purdue.edu/ gfrancis/Publications/BackwardMasking/. The simulations can be run in a Web browser that supports the Java programming language. This article describes the motivation for making the simulations available and gives a brief introduction as to how the simulations are used. The source code is available on the Web page, and this article describes how the code is organized. PMID- 14748496 TI - Computing measures of simplicity of fit for loadings in factor-analytically derived scales. AB - A very simple structure is sought when factor analysis is used to develop measurement scales. The SIMLOAD program computes measures of factorial simplicity for rows and columns of loading matrices (usually the factor pattern) as well as some overall measures. These include Kaiser's (1974) index of factorial simplicity for variables (rows), the author's scale fit index for factors (columns), Bentler's (1977) scale-free matrix measure, and hyperplane counts. Routine use of these measures is recommended for multifactor scale development. The measures may also be useful in more general factor applications and in confirmatory as well as exploratory analyses. SIMLOAD also computes factor scale intercorrelations, scale alpha coefficients (including alpha when an item is removed), and sorted loadings for ease of interpretation. PMID- 14748497 TI - GGUMLINK: a computer program to link parameter estimates of the generalized graded unfolding model from item response theory. AB - The generalized graded unfolding model (GGUM) is an item response theory (IRT) model that implements symmetric, nonmonotonic, single-peaked item characteristic curves. The GGUM is appropriate for measuring individual differences for a variety of psychological constructs, especially attitudes. Like other IRT models, the location and scale (i.e., the metric) of parameter estimates from the GGUM are data dependent. Therefore, parameter estimates from alternative calibrations will generally not be comparable, even when responses to the same items are analyzed. GGUMLINK is a computer program developed to reexpress parameter estimates from two separate GGUM calibrations in a common metric. In this way, the results from separate calibrations of model parameters can be compared. GGUMLINK can secure a common metric by using one of five methods that have recently been generalized to the GGUM. The GGUMLINK executable program is available free and may be downloaded from http://www.education.umd.edu/EDMS. PMID- 14748498 TI - Estimation of the MIRID: a program and a SAS-based approach. AB - The MIRID CML program is a program for the estimation of the parameter values of two different componential IRT models: the Rasch-MIRID and the OPLM-MIRID (Butter, 1994; Butter, De Boeck, & Verhelst, 1998). To estimate the parameters of both models, the program uses a CML approach. The model parameters can also be estimated with a MML approach that can be implemented in PROC NLMIXED of SAS Version 8. Both the MIRID CML program and the MML SAS approach are explained and compared in a simulation study. The results showed that they did about equally well in estimating the values of the item parameters but that there were some differences in the estimation of the person parameters, as could be expected from the differential assumptions regarding the distribution of the persons. The SAS MML approach is much slower than the MIRID CML program, but it is more flexible. PMID- 14748499 TI - The reliability and stability of verbal working memory measures. AB - The psychometric properties of several commonly used verbal working memory measures were assessed. One hundred thirty-nine individuals in five age groups (18-30, 50-59, 60-69, 70-79, and 80+ years) were tested twice (Time I and Time II) on seven working memory span measures (alphabet span, backward digit span, missing digit span, subtract 2 span, running item span, and sentence span for syntactically simple and complex sentences), with an interval of approximately 6 weeks between testing. There were significant effects of age on all but two of the tasks. All the measures had adequate internal consistency. Correlations between performances at Time I and Time II were significant for all the tasks, other than the missing digit span task. The magnitude of the correlations was similar across the age groups and ranged from .52 to .81. Classification of subjects into discrete memory span groups on the basis of a single measure was highly inconsistent across testing sessions and tasks. Classification into upper and lower quartiles was more stable than using a cutoff score for group membership or than classification into high-, medium-, and low-span groups. Correlational analyses showed that there was a moderate relationship between performances on many of the span tasks. Confirmatory factor analysis suggested that six of the seven tasks reflected a common factor. Both test-retest reliability and stability of classification improved when a composite measure reflecting performance on several tasks was used. PMID- 14748500 TI - PsyScript: a Macintosh application for scripting experiments. AB - PsyScript is a scriptable application allowing users to describe experiments in Apple's compiled high-level object-oriented AppleScript language, while still supporting millisecond or better within-trial event timing (delays can be in milliseconds or refresh-based, and PsyScript can wait on external I/O, such as eye movement fixations). Because AppleScript is object oriented and system-wide, PsyScript experiments support complex branching, code reuse, and integration with other applications. Included AppleScript-based libraries support file handling and stimulus randomization and sampling, as well as more specialized tasks, such as adaptive testing. Advanced features include support for the BBox serial port button box, as well as a low-cost USB-based digital I/O card for millisecond timing, recording of any number and types of responses within a trial, novel responses, such as graphics tablet drawing, and use of the Macintosh sound facilities to provide an accurate voice key, saving voice responses to disk, scriptable image creation, support for flicker-free animation, and gaze-dependent masking. The application is open source, allowing researchers to enhance the feature set and verify internal functions. Both the application and the source are available for free download at www.maccs.mq.edu.au/-tim/psyscript/. PMID- 14748501 TI - TPL-KATS-card sort: a tool for assessing structural knowledge. AB - The study of how individuals organize knowledge has been a popular endeavor for several decades. As a result, techniques have been developed to assess how individuals represent and organize knowledge internally. Although several conceptual knowledge elicitation methods have been developed and used to assess the organization of knowledge, their use is often labor intensive and time consuming. Presented here is a software tool that was developed to reduce the problems associated with manually administering the conceptual knowledge elicitation technique, or card sorting. The TPL-KATS-card sort software not only simplifies the administration of the task, but also adds features to the card sorting task such as media insertion, time stamping, and instructorless administration. In the present article, an introduction to the card-sorting technique is provided, the new software tool is described, and the advantages of the software are detailed. PMID- 14748502 TI - RETR_PWR: an SAS macro for retrospective statistical power analysis. AB - In contrast to prospective power analysis, retrospective power analysis provides an estimate of the statistical power of a hypothesis test after an investigation has been conducted rather than before. In this article, three approaches to obtaining point estimates of power and an interval estimation algorithm are delineated. Previous research on the bias and sampling error of these estimates is briefly reviewed. Finally, an SAS macro that calculates the point and interval estimates is described. The macro was developed to estimate the power of an F test (obtained from analysis of variance, multiple regression analysis, or any of several multivariate analyses), but it may be easily adapted for use with other statistics, such as chi-square tests or t tests. PMID- 14748503 TI - Comparison of video- and EMG-based evaluations of the magnitude of children's emotion-modulated startle response. AB - We investigated the reliability and validity of a video-based method of measuring the magnitude of children's emotion-modulated startle response when electromyographic (EMG) measurement is not feasible. Thirty-one children between the ages of 4 and 7 years were videotaped while watching short video clips designed to elicit happiness or fear. Embedded in the audio track of the video clips were acoustic startle probes. A coding system was developed to quantify from the video record the strength of the eye-blink startle response to the probes. EMG measurement of the eye blink was obtained simultaneously. Intercoder reliability for the video coding was high (Cohen's kappa = .90). The average within-subjects probe-by-probe correlation between the EMG- and video-based methods was .84. Group-level correlations between the methods were also strong, and there was some evidence of emotion modulation of the startle response with both the EMG- and the video-derived data. Although the video method cannot be used to assess the latency, probability, or duration of startle blinks, the findings indicate that it can serve as a valid proxy of EMG in the assessment of the magnitude of emotion-modulated startle in studies of children conducted outside of a laboratory setting, where traditional psychophysiological methods are not feasible. PMID- 14748504 TI - Surreptitiously projecting different movies to two subsets of viewers. AB - A new technique (manipulation of overlapping rivalrous images by polarizing filters, or MORI) has been invented for presenting on the same screen two different images that can be seen separately by two groups of viewers without their noticing the overlap. It can easily create desired artificial conflicts among viewers. Two perpendicular polarizing filters provide separate invisible channels from dual video projectors to two groups of viewers on a single screen. The basic principle of the presentation technique, details of the apparatus, and limitations are introduced. As an example of the application of this technique, an eyewitness experiment is briefly reported. The results of experiments conducted by the author and colleagues provide evidence of the effectiveness of this technique with various projectors, video materials, group sizes, and ages of participants. PMID- 14748505 TI - Express: a Web-based technology to support human and computational experimentation. AB - Experimental cognitive psychology has been greatly assisted by the development of general computer-based experiment presentation packages. Typically, however, such packages provide little support for running participants on different computers. It is left to the experimenter to ensure that group sizes are balanced between conditions and to merge data gathered on different computers once the experiment is complete. Equivalent issues arise in the evaluation of parameterized computational models, where it is frequently necessary to test a model's behavior over a range of parameter values (which amount to between-subjects factors) and where such testing can be speeded up significantly by the use of multiple processors. This article describes Express, a Web-based technology for coordinating "clients" participants or computational models) and collating client data. The technology provides an experiment design editor, client coordination facilities (e.g., automated randomized assignment of clients to groups so that group sizes are balanced), general data collation and tabulation facilities, a range of basic statistical functions (which are constrained by the specified experimental design), and facilities to export data to standard statistical packages (such as SPSS). We report case studies demonstrating the utility of Express in both human and computational experiments. Express may be freely downloaded from the Express Web site (http://express.psyc.bbk.ac.uk/). PMID- 14748506 TI - Validating internet research: a test of the psychometric equivalence of internet and in-person samples. AB - This study evaluated the psychometric equivalency of Web-based research. The Sexual Boredom Scale was presented via the World-Wide Web along with five additional scales used to validate it. A subset of 533 participants that matched a previously published sample (Watt & Ewing, 1996) on age, gender, and race was identified. An 8 x 8 correlation matrix from the matched Internet sample was compared via structural equation modeling with a similar 8 x 8 correlation matrix from the previously published study. The Internet and previously published samples were psychometrically equivalent. Coefficient alpha values calculated on the matched Internet sample yielded reliability coefficients almost identical to those for the previously published sample. Factors such as computer administration and uncontrollable administration settings did not appear to affect the results. Demographic data indicated an overrepresentation of males by about 6% and Caucasians by about 13% relative to the U.S. Census (2000). A total of 2,230 participants were obtained in about 8 months without remuneration. These results suggest that data collection on the Web is (1) reliable, (2) valid, (3) reasonably representative, (4) cost effective, and (5) efficient. PMID- 14748507 TI - Timed picture naming: extended norms and validation against previous studies. PMID- 14748508 TI - Normative data for 144 compound remote associate problems. AB - We have developed and tested 144 compound remote associate problems. Across eight experiments, 289 participants were given four time limits (2 sec, 7 sec, 15 sec, or 30 sec) for solving each problem. This paper provides a brief overview of the problems and normative data regarding the percentage of participants solving, and mean time-to-solution for, each problem at each time limit. These normative data can be used in selecting problems on the basis of difficulty or mean time necessary for reaching a solution. PMID- 14748509 TI - NFAT transcription factors control HIV-1 expression through a binding site downstream of TAR region. AB - NFAT factors control HIV-1 transcription. We show here that, in addition to binding to two NF-kappaB/NFAT sites within the U3 HIV LTR, NFATc1 and NFATc2 bind to an NFAT site within the LTR's U5 region. Mutations in this site which abolish NFAT binding reduce the ability of NFATs to transactivate LTR-mediated transcription. Mutations in all three NFAT sites strongly interfered with LTR induction, but affected moderately the stimulatory effect of Tat. PMID- 14748510 TI - Frequency of Valpha24+CD161+ natural killer T cells and invariant TCRAV24-AJ18 transcripts in atopic and non-atopic individuals. AB - Th2 cells play a central role in type I allergies. However, the source of interleukin-4 which may lead to a Th1/Th2 imbalance is unknown. Valpha24+CD161+ Natural killer T (NKT) cells secrete high amounts of interleukin-4 and/or interferon-gamma and are assumed to participate in the initiation of Th1/Th2 immune responses. Their contribution to the development of Th2-dependent type I allergies is controversial. Our objective in this paper was to determine whether Valpha24+CD161+ NKT cells differ in atopic and non-atopic adults. Venous blood was obtained from thirteen atopic and sixteen healthy adult probands. Valpha24+CD161+ NKT cells were determined in CD4+, CD8(bright/dim) and CD4-CD8- lymphocytes by flow cytometry. At the molecular level, the amounts of T cell receptor (TCR) AV24-AJ18 transcripts were quantified with respect to TCRAV24 chain transcripts alone or to all TCR alpha chain transcripts. To detect potential inserted nucleotides in the N-region, a novel real-time PCR-based technology was applied. Both CD4+ and CD4-CD8- NKT cells were present at higher frequencies than CD8+ NKT cells in all probands. CD8(dim) NKT cell levels were lower in healthy individuals, although not statistically significantly different to the patients. Amounts of AV24-AJ18 transcripts in relation to total TCR alpha chains and to TCRAV24 alone were equal in both proband groups. N-region diversity was detected in four clones from four different individuals, but altered the amino acid sequence in only one clone of an atopic donor. Analysis of Valpha24+CD161+ NKT cell frequencies at both the cellular and molecular levels failed to reveal significant differences in peripheral blood of atopic and non atopic probands. If NKT cells contribute to development of type I allergies they must do so at earlier times or in other locations. PMID- 14748511 TI - Treatment of experimental sepsis-induced immunoparalysis with TNF. AB - Following a severe septic abdominal infection induced by sublethal cecal ligation and puncture (CLP) in mice, a phase of depressed immune reactivity occurred two days after CLP characterized by a reduced capacity to produce TNF. To determine whether this reduced TNF production causes immunoparalysis as determined by increased susceptibility to bacterial infection and whether therapeutic TNF substitution can be beneficial during this phase, a super-infection with Salmonella enterica Serovar typhimurium or Listeria monocytogenes was induced two days after sublethal CLP. After CLP a state of true immunoparalysis developed during which Salmonella or Listeria super-infection led to increased lethality paralleled by increased bacterial numbers in spleens and livers. Injection of recombinant human TNF before or at the time of super-infection conferred protection to Salmonella but not to Listeria. In the latter case, the infection mortality was even enhanced. Thus, super-infection during the state of sepsis induced immunoparalysis leads to increased lethality. TNF substitution during this state of immunoparalysis can be beneficial or deleterious, depending on the location of TNF activity in the animal, timing of TNF administration, or the type of super-infection. These results demonstrate that impaired TNF production capacity can account for some aspects of immunoparalysis, however, diagnostic parameters are required for a safe TNF substitution therapy. PMID- 14748513 TI - Morphological characterization of mouse B-1 cells. AB - At least three B cell subsets, B-1a, B-1b and B-2 are present circulating peripherally in the mouse. In these animals, B-1 cells constitute a minor fraction of B cells in spleen and are absent in lymph nodes although they represent the main B cell population in peritoneal and pleural cavities. Currently these cells are identified by a surface phenotypic repertoire; they express Mac-1, IgM(high), and B220(low). B-1a cells express CDS. The aim of this work emerged from the fact that the morphology of B-1 cells is not fully characterized. Here we identified B-1 cells using colloidal gold immunocytochemical assays and purified B-1 cells from supernatants of adherent peritoneal cell cultures by a magnetic bead technique. These techniques lead us to demonstrate that, in mice, either B-1a or B-1b cells have a unique morphology distinct from that of B-2 cells. PMID- 14748512 TI - GPI-80, a beta2 integrin associated glycosylphosphatidylinositol-anchored protein, concentrates on pseudopodia without association with beta2 integrin during neutrophil migration. AB - Previously, we identified a glycosylphosphatidylinositol (GPI)-anchored protein, designated GPI-80, present on human neutrophils and monocytes. GPI-80 is physically associated with beta2 integrin on the surface of human neutrophils and may be a regulator of neutrophil adherence and migration. However, it is not yet known how GPI-80 regulates cell adhesion and migration. To investigate the physiological role(s) of GPI-80, we examined the topological relationship of GPI 80 and the beta2 integrin subunit (CD18) on resting and migrating human neutrophils by confocal laser microscopy. On resting neutrophils, GPI-80 was evenly distributed on the cell surface and was associated with CD18. On the other hand, during the early phase of migration (5 - 30 minutes), GPI-80 was detected on cell bodies and also on pseudopodia, but CD18 was detected only on cell bodies, where it was associated with GPI-80. In the late phase of migration (60 minutes), GPI-80 was detected only on pseudopodia and its association with CD18 was hardly observed. Furthermore, some of the GPI-80 on pseudopodia of migrating neutrophils during the late phase was associated with urokinase-type plasminogen activator receptor (uPAR), a regulator of beta2 integrin-dependent adherence and migration. The distribution of GPI-80 on cell surfaces is similar to that of uPAR. These observations suggest that GPI-80 belongs to the beta2 integrin associated GPI-anchored protein family, which has regulatory activity in cell adherence. PMID- 14748514 TI - Lipoteichoic acid from Staphylococcus aureus is a potent stimulus for neutrophil recruitment. AB - Lipoteichoic acid (LTA) is a major immunostimulatory principle of Gram-positive bacteria. Intranasal application of LTA from S. aureus to mice resulted in greatly increased neutrophil and macrophage counts in the bronchoalveolar lavage as well as increased levels of the chemokine KC. The potential of highly pure, bioactive LTA from S. aureus to induce neutrophil recruitment and activation was investigated further in the human system. Although neutrophils expressed the key known receptors, CD14, TLR2 and TLR6, LTA did not induce or prime neutrophils for oxidative burst, or release of chemokines, bactericidal permeability-increasing protein or myeloperoxidase. However, LTA induced a strong release of the chemoattractants LTB4, IL-8, C5a, MCP-1 and the colony-stimulating factor G-CSF in whole blood comparable to stimulation with the same concentration of LPS (S. abortus equi). Further, the cytokine and chemoattractant pattern induced by LTA correlated well with that induced by live S. aureus of the same strain. LTA does not appear to activate neutrophils directly, but is a strong stimulus for the recruitment of phagocytes to the site of infection. PMID- 14748515 TI - Epitope-vaccine strategy against HIV-1: today and tomorrow. AB - Vaccines play important roles in preventing infectious diseases caused by different pathogens. However, some pathogens such as HIV-1 challenge current vaccine strategy. Poor immunogenicity and the high mutation rate of HIV-1 make great difficulties in inducing potent immune responses strong enough to prevent infection via vaccination. Epitope-vaccine, which could intensively enhance predefined epitope-specific immune responses, was suggested as a new strategy against HIV-1 and HIV-1 mutation. Epitope-vaccines afford powerful approaches to elicit potent, broad and complete immune protection against not only primary homologous viral isolates but also heterologous viral mutants. Although most studies are still preliminary now, epitope-vaccine as a novel strategy against the AIDS epidemic has great developmental potential. To trigger T-cell-dependent IgG antibody responses and improve affinities of the epitope-specific antibodies, approaches such as recombinant multi-epitope-vaccination and prime-boosting vaccination were suggested. Cellular immune responses, especially CTL responses, could also be elicited and enhanced in addition to humoral immune responses. Developed epitope-vaccines activating both arms of the immune system would benefit prevention and immunotherapy not only against HIV but also other chronic infections. PMID- 14748516 TI - Advance care planning redux: it's time to talk. PMID- 14748517 TI - Differential advantage and hospice care. PMID- 14748518 TI - Social invitations, time constraints, and professional distance. PMID- 14748519 TI - Palliative care at the end of life: comparing quality in diverse settings. AB - There is growing awareness that pain and other symptoms are often poorly managed at the end of life. The purpose of this quality improvement project was to compare the quality of care provided to a convenience sample of 195 patients who died during a six-month period, using a retrospective chart review. Quality was defined by symptom documentation, use of diagnostic and therapeutic procedures in the final 48 hours of life, and determination of advance directives. Daily and total charges incurred by these patients were also captured. Symptom distress was common, and diagnostic and therapeutic procedures were widespread. These data suggest areas for improvement in clinical practice, in palliative care units, and in all settings where end-of-life care is provided. Also, the data can guide future research into the quality of care provided to dying persons. PMID- 14748520 TI - Cultural competence in hospice. AB - Research shows that ethnic minorities access hospice care significantly less often than Caucasians. In part, this has been attributed to the lack of cultural competence among hospice staff. To assess cultural competence among hospice workers, this article evaluates the results of a descriptive, exploratory survey that was submitted to 125 interdisciplinary hospice employees and completed by 113 of those employees. Cultural-competence behavior scores varied widely based on two factors: increased education and previous cultural-diversity training. The results of this study provide information regarding cultural competence in hospice. In their efforts to provide culturally appropriate end-of-life care, hospices can use the information in this study to implement intervention plans aimed at increasing cultural competence among hospice staff. PMID- 14748521 TI - Sexuality at the end of life. AB - There is very little research literature that addresses sexuality at the end of life. Although end-of-life care has become a priority for nursing education, the issue of end-of-life sexuality is not included in the curriculum. Nurses are frequently in a position to establish relationships with couples that encourage a frank discussion and information sharing. As patient advocates, nurses can address end-of-life sexuality issues by taking a sexual history and implementing a general intervention model, such as the PLISSIT. Couples need to be reassured that if they have enjoyed a close sexual relationship, sexual intimacy may continue to be part of their relationship, even at the end of life. PMID- 14748522 TI - Educational needs of hospice social workers: spiritual assessment and interventions with diverse populations. AB - Based on a national survey, this study analyzes the roles and educational needs of hospice social workers regarding assessment and intervention in spirituality, religion, and diversity of their patients. Sixty-two social workers responded to the survey. Results suggest that spiritual care is shared among hospice team members and that most social workers feel comfortable in addressing these issues. However, role conflict and role ambiguity also exist. Respondents to the survey often felt ill-prepared to deal with some complex faith-based conflicts related to diversity. They saw themselves in need of assessment models and end-of-life decision-making interventions regarding assisted suicide and euthanasia. This study provides recommendations for social work practice, education, and research. PMID- 14748523 TI - Acute pain in advanced cancer: an opioid dosing strategy and illustration. AB - Opioid dosing strategies for acute pain differ from strategies for chronic pain management. The basic principles of effective, safe dosing are rapid titration to the onset of analgesia followed by maintenance infusions based upon the titrated dose. This article presents guidelines and case histories for safe and effective dosing. PMID- 14748524 TI - Current use of guidelines, protocols, and care pathways for symptom management in hospice. AB - Evidence-based guidelines or care pathways for symptom management could provide a means to reduce symptom distress in dying patients. We surveyed directors of nursing from hospices affiliated with the Population-based Palliative Care Research Network (PoPCRN) regarding their hospices' current use of and attitudes toward written symptom management materials. A majority (53/78, 68 percent) of participating hospices reported use of written materials, such as guidelines, protocols, or care pathways, for one or more symptoms. Materials were based on multiple sources and varied from simple medication orders to more comprehensive, multicategory symptom management resources. Regardless of the composition, these materials were perceived as helpful. Given this favorable view, variations in the use and content of written materials may signify an opportunity to decrease symptom distress in hospice through the implementation of evidence-based symptom management resources. PMID- 14748525 TI - Intravesical lidocaine for formalin-induced suprapubic pain: case report. AB - This case report describes the management of intravesical formalin induced suprapubic pain using intravesical lidocaine. PMID- 14748526 TI - Discontinuing life support: whose call? PMID- 14748527 TI - Annie's song: a student's reflection on a memorable patient's end-of-life care. PMID- 14748528 TI - The meaning of curriculum: content to be covered or stories to be heard? PMID- 14748529 TI - "Covering content" and teaching thinking: deconstructing the additive curriculum. AB - For more than 25 years, reliance on conventional pedagogy has led nurse educators to persistently focus on what students need to learn to enter contemporary practice settings. Therefore, as biomedical and nursing knowledge grows and the health care system in which students will practice becomes increasingly complex, content is persistently added to nursing curricula, while little is taken out. An underlying assumption of this approach is that if important content is "covered," thinking necessarily follows. This study, using Heideggerian hermeneutics, examines the relationship between covering content and thinking by explicating the common experiences of teachers enacting interpretive pedagogies. One of the themes that emerged from this analysis is presented: "Covering Content" and Teaching Thinking: Deconstructing the Additive Curriculum. PMID- 14748530 TI - Baccalaureate nursing students' attitudes toward poverty: implications for nursing curricula. AB - Given the link between poverty and health, nurses, in their work in hospitals and in the community, often come into contact with people who are poor. To be effective care providers, nurses must have an adequate understanding of poverty and a positive attitude toward people who are poor. This study examined attitudes toward poverty among baccalaureate nursing students (N = 740) at three Canadian universities. Students' attitudes were neutral to slightly positive. Personal experiences appeared to have an important influence on the development of favorable attitudes. The findings point to several considerations for nursing curricula. Students should not only be provided with classroom opportunities for critical exploration of poverty and its negative effects on individuals and society, but also have clinical learning experiences that bring them face-to-face with people who are poor, their health concerns, and the realities of their circumstances. Thoughtful critique of poverty-related issues and interpersonal contact may be effective strategies to foster attitude change. PMID- 14748531 TI - Analysis of spirituality content in nursing textbooks. AB - Although most nurses believe spiritual care is an integral component of quality, holistic nursing care, they rarely address spiritual issues and typically feel unprepared to do so. One reason for nurses' lack of preparedness to provide spiritual interventions is that their basic education only minimally discusses spirituality and related issues. This is compounded by the problem that only sporadic reference to spiritual care is found in most nursing textbooks. This study was conducted to analyze the content related to spirituality in nursing textbooks in order to determine where spiritual care is addressed and evaluate its adequacy. A total of 50 textbooks from a wide variety of nursing specialty areas were selected from the most recent Brandon Hill list. These books were examined to assess the percentage of pages discussing spiritual issues and analyze inclusion of core content essential for nursing practice. Although there was considerable variation among the books from all specialty areas, overall, hospice/terminal care, fundamentals of nursing, health assessment/health promotion, and transcultural nursing textbooks provided the most information about spirituality and spiritual care. Textbooks focusing on professional issues, medical-surgical nursing, maternal-child health nursing, critical care nursing, and community health nursing contained the least spiritual content. Suggestions are made regarding how to integrate spiritual issues and spiritual care in all nursing textbooks that pertain directly to patient care. PMID- 14748532 TI - A facilitative approach to learning about curriculum development. AB - Graduate students have high ambitions and desire excellence in their work. Creating learning opportunities that capture this drive and help them achieve and exceed their goals is a challenge for educators. This article describes two teaching approaches, group process and an adaptation of Bensusan's escalator model, which were used in a graduate nursing course to help students learn about curriculum development. Students participated as a faculty group, submitting successive iterations of their work as they developed hypothetical curricula. Benefits students identified from course faculty's critiques of their submissions included experiencing enhanced self-direction, self-esteem, and mutual respect among students and between students and course faculty, as well as authentic curriculum development in a safe, caring, and supportive context. This article discusses the strengths and limitations of this pragmatic and productive learning approach to preparing future nurse educators for their role as curriculum developers. PMID- 14748533 TI - Students' self-identified learning needs: a case study of baccalaureate students designing their own death and dying course curriculum. AB - The knowledge needed to provide competent care to dying clients and their families and to meet the established criteria for effective death and dying curricula was explored by junior-level and senior-level baccalaureate honors students, who identified their own learning needs and resources. After completing the self-designed curriculum, student-identified learning needs were compared to the American Association of Colleges of Nursing competencies for providing high quality end-of-life care. Analysis of the student-identified objectives revealed a high level of congruity with the nationally established competencies. PMID- 14748534 TI - Instilling the value of reading literature through student-led book discussion groups. AB - With the desire to instill the value of lifelong reading, nursing faculty and a librarian developed a student-led book discussion group as an innovative teaching strategy for a senior seminar course. Inclusion of the librarian was unique and influenced the shape and rigor of the assignment. In this assignment, students chose one of the faculty-selected books, researched its author(s), read relevant professional book reviews, and developed questions for their peer discussion groups. Student and faculty responses were positive and clearly demonstrated the benefits of this assignment for students' personal development and professional growth. PMID- 14748535 TI - Case management in the undergraduate curriculum. AB - The redesign of an undergraduate curriculum at one baccalaureate nursing program resulted in adoption of a required course on case management for chronic illnesses. The course is structured around rehabilitation standards and medical surgical and community health principles of nursing care. A service-learning model was used to develop the clinical experience, which involved student nurses visiting the homes of individuals with chronic illnesses. These visits were made in collaboration with personnel from community-based agencies or programs. This article discusses the design of the course and some of the content areas and clinical activities. PMID- 14748536 TI - NMR studies of structure and function of biological macromolecules. PMID- 14748537 TI - Successful reconstruction of damaged ocular outer surface in humans using limbal and conjuctival stem cell culture methods. AB - When the ocular outer surface is badly damaged, subsequent corneal transplantation fails due to the absence of basal cells that are needed to support the graft. With the realization that the limbus and the conjunctiva have adult stem cells that can be cultured, it has been possible for us to explant culture these on de-epithelized human amniotic membrane, and to graft the resulting viable and transparent epithelium to 125 needy human patients with success. Ultrastructural, histological, biochemical and immunological assays establish the identity of the cells and the tissue formed. PMID- 14748538 TI - Solubilization of adenylyl cyclase from human myometrium in a alphas-coupled form. AB - Adenylyl cyclase (AC) was extracted from human myometrium with either non-ionic (Lubrol-PX or Triton X-100) or zwitterionic (3-[3 cholamidopropyl)dimethylammonio]-1-propanesulfonate, CHAPS) detergents. The soluble enzyme was stimulated by forskolin, a hydrophobic activator, in the presence of Mg2+ indicating that the catalytic subunit had not been damaged after solubilization. The enzyme was also activated by 5'-guanylyl imidodiphosphate (Gpp(NH)p) showing that the catalytic unit was not separated from stimulatory guanine nucleotide binding protein (Gs) during the extraction. Both activators showed different effects on the stimulatory efficacy and potency of AC activity solobulized with detergents. Gel filtration of Lubrol-PX and CHAPS extracts over a Sepharose CL-2B column partially resolved AC and its complexes. The chromatographic profile for Lubrol-solubilized AC presented a main peak of about 200 kDa whereas CHAPS-solubilized AC showed a dominant peak of about 1100 kDa. The heterodisperse peaks obtained revealed that the catalytic AC subunit was not separated from Gs proteins after gel filtration, and that AC could be associated with other cellular proteins. When Lubrol extract was submitted to anionic exchange chromatography, the enzyme was purified about 7.5 fold (enzymatic activity of 48.1 pmol/min/mg of protein). The catalytic subunit was co-eluted with both AC-activating proteins Galphas large (52.2 kDa) and Galphas small (48.7 kDa). This is the first demonstration of the stable physical association of AC with both alphas subunits of G proteins in human myometrium. PMID- 14748539 TI - Small heat shock protein hsp27 as a possible mediator of intercellular adhesion induced drug resistance in human larynx carcinoma HEp-2 cells. AB - The confluence-dependent resistance of human larynx carcinoma HEp-2 cells to hydrogen peroxide and a new antitumor drug based on the combination of vitamins C and B12b was studied. It was found that this resistance in growing cells is suppressed by the disruption of intercellular contacts by EGTA and is related neither to the activity of P-glycoprotein nor to the content of intracellular glutathione and the activities of glutathione S-transferases, glutathione peroxidase and glutathionine reductase. Here we showed that the level of expression of the small heat shock protein hsp27, which is known to protect cells from a variety of stresses associated with apoptosis, in growing confluent cells both in the presence and absence of the vitamins B12b and C is much higher (about 20-25 times) than in non-confluent cells. Taken together, the results suggest that the confluence-dependent resistance of cells to the combination of vitamins C and B12b and to hydrogen peroxide is mediated by hsp27 overexpression, which is activated via cell-cell adhesion. PMID- 14748540 TI - The mutation in the N-terminal domain of RGS4 disrupts PA-conferred inhibitory effect on GAP activity. AB - Regulator of G protein signaling (RGS) proteins are GTPase-activating proteins (GAP) for G protein alpha-subunits and are thought to be responsible for rapid deactivation of G protein mediated signaling pathway. In this present study, we demonstrate that PA is the most efficient candidate to inhibit GAP activity of RGS4. The functional significance of N-terminus of RGS4 in respose to PA-granted inhibition on GAP activity has been studied with the site mutation in the N terminus of RGS4. These site-directed mutations in the N-terminal domain do not severely disrupt its association with liposomes of PA. However, RGS4L23E diminishes the inhibition of GAP activity by PA compared with the wild type RGS4, whereas RGSR22E abrogates the inhibitory effect by PA on GAP activity. The correspondent conformational discrepancy in the RGS domain of these mutants in the presence of PA vesicles was detected from fluorescence experiments. It is suggested that the functional pertinence between the N-terminus and RGS domain may be important to modulate PA-conferred inhibitory effect on its GAP activity. PMID- 14748541 TI - Biomechanics and neuroscience: a failure to communicate. AB - This commentary is the third in a series commemorating the 50th anniversary of the American College of Sports Medicine. The charge to the commentators was to provide insight on the origins and directions in the fields of physical activity and disease prevention (Haskell), applied exercise physiology (Wilmore), biomechanics and neuroscience (Enoka), and the physiology of exercise (Holloszy). In contrast to the innovation and vitality that characterizes the activities of the College in most of these fields, the inclusion of biomechanics and neuroscience as essential elements in realizing the mission of the College has been much less impressive. What we have here, as the saying goes, is a failure to communicate. PMID- 14748542 TI - Integration of metabolic and mitogenic signal transduction in skeletal muscle. AB - Exercise has diverse effects on metabolic and mitogenic signaling pathways in human skeletal muscle, implying specificity of intracellular signaling cascades. The role of several parallel signaling cascades are discussed in an effort to assign a physiological role for these targets in the regulation of exercise mediated responses on metabolism and gene expression. PMID- 14748543 TI - Why rest stimulates bone formation: a hypothesis based on complex adaptive phenomenon. AB - Moderate exercise is an ineffective strategy to build bone mass. The authors present data demonstrating that allowing bone to rest between each load cycle transforms low- and moderate-magnitude mechanical loading into a signal that potently induces bone accretion. They hypothesize that the osteogenic nature of rest-inserted loading arises by enabling osteocytes to communicate as a small world network. PMID- 14748544 TI - Contraction-induced oxidants as mediators of adaptation and damage in skeletal muscle. AB - Contracting skeletal muscle generates reactive oxygen and nitrogen species (ROS) that can induce changes in gene expression or cell damage depending upon the pattern of production and the endogenous protective systems. The hypothesis is presented that skeletal muscle uses contraction-induced ROS as signals to induce adaptive responses including maintenance of oxidant homeostasis and prevention of oxidative damage. PMID- 14748545 TI - Aldose reductase: a key player in myocardial ischemic injury. AB - In the search for increasing effectiveness of reperfusion therapy, the authors demonstrate that the polyol pathway enzyme aldose reductase is a key component of myocardial ischemic injury and that inhibitors of this enzyme limit ischemic injury and improve functional recovery on reperfusion. PMID- 14748546 TI - Vulnerability of pulmonary capillaries during exercise. AB - Pulmonary capillaries have a dilemma. Their walls must be extremely thin for efficient gas exchange, but be immensely strong to resist the mechanical stresses that develop during heavy exercise. Elite human athletes at maximal exercise develop changes in the structure of the capillary wall as evidenced by red blood cells in their alveoli. Racehorses routinely break their pulmonary capillaries while galloping. PMID- 14748547 TI - Passive regulation of cardiac output during exercise by the elastic characteristics of the peripheral circulation. AB - A change in cardiac output induced by a change in cardiac performance is accompanied by opposite changes in cardiac filling pressure owing to the resistive and capacitive properties of blood vessels. The inverse relationship between cardiac output and cardiac filling pressure provides a passive (hydraulic) regulatory mechanism that functions to keep cardiac output constant during rest and exercise. PMID- 14748549 TI - Decisions! Decisions! PMID- 14748548 TI - Neurogenesis of excitation-contraction uncoupling in aging skeletal muscle. AB - Excitation-contraction (EC) uncoupling is a major cause of decreased muscle force generating capacity (specific force). However, the underlying mechanisms of EC uncoupling in muscle from aging mammals have not been characterized. We propose that impaired motor neuron function with aging leading to muscle denervation, a process probably initiated earlier than detected by in vitro morphologic techniques, results in EC uncoupling. PMID- 14748550 TI - Performance of musicians and nonmusicians on dichotic chords, dichotic CVs, and dichotic digits. AB - Perception of dichotic chords (free recall and directed recall), nonsense syllables (CVs), and three-pair digits was assessed on 24 musicians and 24 nonmusicians. On the dichotic-CV and dichotic-digit free-recall tasks, there was a significant right-ear advantage, but there were no group differences. With the dichotic-chords, free-recall condition, a significant left-ear advantage was observed but no group difference. For the dichotic-chords, directed-recall conditions, the musicians performed significantly better by 10 percent than the nonmusicians. Unexpectedly, for the dichotic chords, the 62-72 percent correct performances were better on the free-recall condition than the 42-55 percent performances on the directed-recall conditions. These differences between the two response modes were attributed to the difficulty of the dichotic-chord listening tasks and the probabilities associated with the closed-set response paradigms. The findings suggest that the dichotic-chord paradigm used in this study should not be included in clinical protocols used to assess auditory perceptual abilities. PMID- 14748551 TI - Should significant others be encouraged to join adult group audiologic rehabilitation classes? AB - The benefit of participation in group audiologic rehabilitation classes was examined for adults with hearing loss (subjects) and their significant others (SOs). Thirteen subjects attended the classes with their SOs, and 12 subjects attended the classes on their own. All subjects attended six 90-minute classes consisting of informational lectures, and training in communication strategies, auditory perception, and auditory and visual perception. Self-assessment scales measuring hearing aid benefit and use of communication strategies were completed prior to class participation and following the completion of all classes. The results indicated that the majority of subjects reported increased use of communication strategies following class participation. In addition, a significant reduction in hearing handicap following class participation was measured across all subjects and SOs, and the greatest reduction in handicap was measured for subjects who attended the classes with their SOs. SO participation in group AR (audiologic rehabilitation) classes should be encouraged. PMID- 14748552 TI - Normal brief-tone bone-conduction behavioral thresholds using the B-71 transducer: three occlusion conditions. AB - Behavioral thresholds were measured from 31 adults with normal hearing for 500, 1000, 2000, and 4000 Hz brief tones presented using a B-71 bone oscillator. Three occlusion conditions were assessed: ears unoccluded, one ear occluded, and both ears occluded. Mean threshold force levels were 67, 54, 49, and 41 dB re:1 microN peak-to-peak equivalent in the unoccluded condition for 500, 1000, 2000, and 4000 Hz, respectively (corrected for air-conduction pure-tone thresholds). A significant occlusion effect was observed for 500 and 1000 Hz stimuli. These thresholds may be used as the 0 dB nHL (normal-hearing level) for brief-tone bone conduction stimuli for auditory brainstem response testing. PMID- 14748553 TI - Transient evoked otoacoustic emissions in adults: a comparison between two test protocols. AB - This study compares the performance of the Quickscreen and Default protocols of the ILO-96 Otodynamics Analyzer in recording transient evoked otoacoustic emissions (TEOAEs) from adults using clinical decision analysis. Data were collected from 25 males (mean age = 29.0 years, SD = 6.8) and 35 females (mean age = 28.1 years, SD = 9.6). The results showed that the mean signal-to-noise ratios obtained from the Quickscreen were significantly greater than those from the Default protocol at 1, 2, and 4 kHz. The comparison of the performance of the two protocols, based on the results using receiver operating characteristics curves, revealed a higher performance of the Quickscreen than the Default protocol at 1 and 4 kHz but not at 2 kHz. In view of the enhanced performance of the Quickscreen over the Default protocol in general, the routine use of the Default protocol for testing adults in audiology clinics should be reconsidered. PMID- 14748554 TI - Word recognition and the articulation index in older listeners with probable age related auditory neuropathy. AB - This retrospective analysis of existing data was derived from 957 members of a population-based cohort who participated in a prior study of the prevalence of central auditory dysfunction. Word recognition scores (WRS) at three intensity levels were compared to predicted scores based on the Articulation Index (AI) and the Thornton-Raffin 95 percent critical differences. In 112 (11.7 percent) participants, one or more word recognition scores were significantly below the predicted score, which we consider a subtle sign of possible auditory neuropathy. In contrast, classic signs of retrocochlear dysfunction were found in only three people (0.3 percent) using rollover of the performance-intensity function for phonetically balanced word lists, in two (0.2 percent) people using the guideline of Yellin et al (1989), and in 54 people (5.6%) using a 20-point difference between the AI (x 100) and the WRS. Subtle signs of possible auditory neuropathy were more frequent than the classic signs. Comparing WRS at several high presentation levels to the AI is suggested as a method to screen for subtle neuropathy. Elderly listeners whose WRS fall below the Thornton-Raffin 95 percent critical difference based on AI should be considered for further testing for age related auditory neuropathy. PMID- 14748555 TI - The effects of frequency response on speech perception for cochlear implant users. AB - The aim of this study was to investigate the effects on speech perception of manipulating filter gains in a cochlear implant speech processor. Five implantees, who use the CI22 implant and Spectra processor manufactured by Cochlear Ltd, participated. Four experimental maps were created that were identical to their clinical map except for the profile of gains across the filters. Experimental gain profiles had rising or falling gains across the frequency range, or emphasized or de-emphasized the middle frequencies, relative to the clinical map. Perception of CNC (consonant-vowel-consonant) words at 70 dB SPL was significantly better with the clinical map than with all experimental maps, whereas at the lower level (60 dB SPL) there was minimal difference between the maps, with the low-frequency emphasis map giving significantly better scores than the high-frequency emphasis map. Perception of sentences at 70 dB SPL with a signal-to-noise ratio of +10 dB was better with the high-frequency emphasis map than with the low-frequency emphasis map. None of these best-conditions, however, were statistically better than the clinical map. The results highlighted the importance of signal audibility for speech perception with cochlear implants. PMID- 14748556 TI - The effects of toluene on the central nervous system. AB - In recent decades the organic solvent toluene (methylbenzene) has emerged as one of the best-studied neurotoxins. Long-term and intense exposure to toluene vapors in humans who abuse spray paint and related substances has led to the recognition that toluene has a severe impact on central nervous system myelin. Chronic toluene abuse produces a devastating neurological disorder, of which dementia is the most disabling component. The clinical syndrome, toluene leukoencephalopathy, can be detected by a combination of characteristic symptoms and signs, detailed neurobehavioral evaluation, and brain magnetic resonance imaging. In this paper, we consider the impact of toluene abuse on our society, describe the specific neurobehavioral deficits in toluene leukoencephalopathy, review the spectrum of neuroimaging findings in patients with this disorder, summarize the teratogenic effects of toluene in both humans and animal models, and offer possible explanations for the range of neuropathological damage seen in brains of individuals who chronically abuse toluene. PMID- 14748557 TI - Differential expression of sst1, sst2A, and sst3 somatostatin receptor proteins in low-grade and high-grade astrocytomas. AB - We have previously reported that sst2A somatostatin receptors are frequently overexpressed in human meningiomas. Initial clinical observations suggest that somatostatin analogues may also be of value for imaging and treatment of other human intracranial tumors, including astrocytomas. However, contradictory results have been reported regarding the expression of somatostatin receptors in low grade and high-grade astrocytomas. Therefore, we determined the precise pattern of somatostatin receptor protein expression in 8 diffuse astrocytoma (DA), 10 anaplastic astrocytomas (AA), and 32 glioblastoma multiforme (GBM) using immunohistochemistry and Western blot analysis. sst1 and sst2A somatostatin receptors were not present in DA and only occasionally detected in AA. In GBM, sst1 was present in 66%, and sst2A was found in 44% of the tumors. sst3 receptors were present in 38% of DA, 40% of AA, and 84% of GBM. Thus, loss of differentiation was significantly associated with increased expression of sst1, sst2A, and sst3 somatostatin receptors. In contrast, sst4 and sst5 receptors were found in 80% and 25% of all cases, respectively, in a manner independent of histological grade. No significant correlation was found between somatostatin receptor expression and the proliferation rate of the tumors as determined by MIB I immunostaining. Furthermore, the presence or absence of the 5 somatostatin receptor subtypes did not significantly influence survival time in 14 GBM patients. PMID- 14748558 TI - Intralaminar thalamic nuclei lesions: widespread impact on dopamine denervation mediated cellular defects in the rat basal ganglia. AB - Intralaminar thalamic nuclei represent a major site of non-dopaminergic degeneration in Parkinson disease, but the impact of this degeneration on the pathophysiological functioning of basal ganglia remains unknown. To address this issue, we compared the effects of 6-hydroxydopamine-induced lesions of nigral dopamine neurons alone or combined with ibotenate-induced lesions of intralaminar thalamic neurons on markers of neuronal metabolic activity in the rat basal ganglia using in situ hybridization histochemistry. Thalamic lesions prevented most of the dopamine denervation-induced changes (i.e. the increases in mRNA levels of enkephalin and GAD67 in the striatum, of GAD67 in the globus pallidus and entopeduncular nucleus, and of cytochrome oxidase subunit-I in the subthalamic nucleus), but did not affect the downregulation of striatal substance P and upregulation of GAD67 in the substantia nigra pars reticulata. We also provide immunohistochemical evidence that thalamic lesions markedly decreased striatal expression of the vesicular glutamate transporter vGluT2, confirming the association of this transporter with the thalamic projections to the basal ganglia. Altogether, these data reveal a major antagonistic influence of thalamic and dopaminergic afferents onto the basal ganglia and suggest that degeneration of thalamic neurons in Parkinson disease may represent an important factor counteracting expression of the defects associated with the dopamine denervation. PMID- 14748559 TI - Immunogold electron microscopy recognizes prion protein-associated particles prepared from scrapie-infected mouse brain. AB - Previous studies have proposed that the disease isoform of prion protein (PrPSc) is particulate. Our purpose was to search by electron microscopy (EM) for particles in fractions of density gradients prepared from differentially centrifuged homogenates of scrapie-infected, normal, and null mouse brain. Only mild detergents were used during the separation process. The low-density fractions derived from scrapie-infected brain were rich in PrP. Three morphologically distinct types of particle were observed. Type 1 particles, measuring approximately 6.8 nm in mean diameter, were found in abundance in the fractions of scrapie-infected brain at the peak PrP concentrations. They were often clumped and adherent to raft-like structures. Type 2 particles, in low density fractions from normal brain, were similar to type 1 but were smaller, with the mean diameter measuring approximately 5.3 nm. Type 3 particles from null brain differed morphologically from types 1 and 2 and were not clumped. The low density of the particles indicated a lipid component, which was confirmed by lipid analysis. Immunogold EM using Mab 6H4 labeled a portion of the particles from scrapie mouse brain, but not those from normal or null brain. Dimensions of PrP suggest that the labeled particles carry a PrPSc dimer per particle. PMID- 14748560 TI - Microglial activation parallels system degeneration in multiple system atrophy. AB - Multiple system atrophy (MSA) is a neurodegenerative disorder that predominantly affects motor-related neuroanatomic structures. The role of microglia in MSA is unknown. To address this issue, we conducted quantitative image studies on the brains from 13 cases of MSA, comprising 8 cerebellar and 5 parkinsonian variants. Microglial and glial cytoplasmic inclusion (GCI) burdens were determined with image analysis on brain sections immunostained with antibodies to HLA-DR and alpha-synuclein. Many activated microglia, as well as GCIs, were noted in motor related structures, including the cerebellar input, extrapyramidal motor, and pyramidal motor structures, but not in the cerebellar output structures. This result indicates that microglial activation, as well as the distribution of GCIs, is system-specific in MSA. The correlation analysis between the microglial and GCI burdens yielded variable yet significant correlations in the cerebellar input, extrapyramidal motor, and pyramidal motor systems, but not in the cerebellar output system. This result suggests that microglial activation is at least partly determined by GCIs or oligodendroglial alpha-synuclein in specific neuroanatomic systems affected in MSA. Taken together, considering the known toxic effects of microglia in neurodegenerative diseases, microglia may play a part in the development of system-specific tissue injuries, contributing to the system-bound clinical and pathological phenotypes. PMID- 14748561 TI - Role of tissue-derived plasminogen activator (t-PA) in an excitotoxic mouse model of neonatal white matter lesions. AB - White matter (WM) lesions in preterm newborns may lead to cerebral palsy. To study WM lesions in a mouse model, we used intrapallial stereotactic injections of ibotenic acid, an N-methyl-D-aspartate receptor agonist. Previous studies support a contribution of tissue-type plasminogen activator (t-PA) to the brain lesions seen in various adult excitotoxic models. Therefore, we studied both 5 day-old (P5) wild-type mice and t-PA knock-out (t-PA-/-) mice. The ibotenic acid doses required to induce WM cysts were lower in the wild-type mice (EC50 < 0.01 microg/animal) than in the t-PA-/- mice (EC50 = 2.5 microg/animal) (p < 0.01), indicating the existence of t-PA-dependent and t-PA-independent mechanisms. Dose dependent prolonged cyst growth occurred in the wild-type mice only. Early microglial activation and astrogliosis were similar in the wild-type and t-PA-/- mice. In adult mice (P45), demyelination occurred at the injection site in both groups but the astroglial scar was denser in the wild-type than in the t-PA-/- mice. These data support involvement of t-PA at several stages of WM lesion formation. Inactivation of t-PA might confer protection by prolonged hemostasis. The role of t-PA in cyst expansion suggests a new approach to the development of neuroprotective strategies in infants with developing WM lesions. PMID- 14748562 TI - Transplanted hematopoietic stem cells from bone marrow differentiate into neural lineage cells and promote functional recovery after spinal cord injury in mice. AB - Recovery in central nervous system disorders is hindered by the limited ability of the vertebrate central nervous system to regenerate lost cells, replace damaged myelin, and re-establish functional neural connections. Cell transplantation to repair central nervous system disorders is an active area of research, with the goal of reducing functional deficits. Recent animal studies showed that cells of the hematopoietic stem cell (HSC) fraction of bone marrow transdifferentiated into various nonhematopoietic cell lineages. We employed a mouse model of spinal cord injury and directly transplanted HSCs into the spinal cord 1 week after injury. We evaluated functional recovery using the hindlimb motor function score weekly for 5 weeks after transplantation. The data demonstrated a significant improvement in the functional outcome of mice transplanted with hematopoietic stem cells compared with control mice in which only medium was injected. Fluorescent in situ hybridization for the Y chromosome and double immunohistochemistry showed that transplanted cells survived 5 weeks after transplantation and expressed specific markers for astrocytes, oligodendrocytes, and neural precursors, but not for neurons. These results suggest that transplantation of HSCs from bone marrow is an effective strategy for the treatment of spinal cord injury. PMID- 14748563 TI - Increased expression of erythropoietin receptor on blood vessels in the human epileptogenic hippocampus with sclerosis. AB - Microvascular (capillary) proliferation is a readily observed, but largely ignored phenomenon of the mesial temporal lobe epilepsy (MTLE) hippocampus. Here, we report that the proliferated capillaries in surgically resected MTLE hippocampi were strongly immunoreactive for erythropoietin receptor (EPO-r). Further, we found that these capillaries were most prominent in areas of the MTLE hippocampus with extensive neuronal loss and gliosis, i.e. the CA3, CA1, and dentate hilus. High-resolution immunogold electron microscopy revealed that the capillary EPO-r was localized to the luminal and abluminal plasma membrane of endothelial cells, to endosome-like structures of these cells, and to pericapillary astrocytic end-feet. Previous studies have shown that systemically administered EPO appears in the cerebrospinal fluid in experimental animals, and the present results are consistent with the idea that EPO enters the brain via receptor-mediated endocytosis. The enrichment of EPO-r shown here suggests a highly efficient uptake of plasma EPO into the MTLE hippocampus and a possible role for this cytokine in epileptogenesis. Moreover, the presence of EPO-r in the MTLE hippocampus may provide a new vehicle for highly efficient delivery of hitherto impermeable drugs into the epileptic brain. PMID- 14748564 TI - SDF-1 (CXCL12) is upregulated in the ischemic penumbra following stroke: association with bone marrow cell homing to injury. AB - The chemokine stromal-derived factor-1 (SDF-1, also known as CXCL12) and its receptor CXCR4 have been implicated in homing of stem cells to the bone marrow and the homing of bone marrow-derived cells to sites of injury. Bone marrow cells infiltrate brain and give rise to long-term resident cells following injury. Therefore, SDF-1 and CXCR4 expression patterns in 40 mice were examined relative to the homing of bone marrow-derived cells to sites of ischemic injury using a stroke model. Mice received bone marrow transplants from green fluorescent protein (GFP) transgenic donors and later underwent a temporary middle cerebral artery suture occlusion (MCAo). SDF-1 was associated with blood vessels and cellular profiles by 24 hours through at least 30 days post-MCAo. SDF-1 expression was principally localized to the ischemic penumbra. The majority of SDF-1 expression was associated with reactive astrocytes; much of this was perivascular. GFP+ cells were associated with SDF-1-positive vessels and were also found in the neuropil of regions with increased SDF-1 immunoreactivity. Most vessel-associated GFP+ cells resemble pericytes or perivascular microglia and the majority of the GFP+ cells in the parenchyma displayed characteristics of activated microglial cells. These findings suggest SDF-1 is important in the homing of bone marrow-derived cells, especially monocytes, to areas of ischemic injury. PMID- 14748565 TI - Adjacent-level disease. PMID- 14748566 TI - Accelerated spondylotic changes adjacent to the fused segment following central cervical corpectomy: magnetic resonance imaging study evidence. AB - OBJECT: The authors studied whether cervical spine motion segments adjacent to a fused segment exhibit accelerated degenerative changes on short-term follow-up magnetic resonance (MR) imaging. METHODS: Preoperative and short-term follow-up (mean duration 17.5 months, range 10-48 months) cervical MR images obtained in 44 patients who had undergone one- or two-level corpectomy for cervical spondylotic myelopathy were evaluated qualitatively and quantitatively. The motion segment adjacent to the fused segment and a segment remote from the fused segment were evaluated for indentation of the thecal sac, disc height, and sagittal functional diameter of the spinal canal on midsagittal T2-weighted MR images. Thecal sac indentations were classified as mild, moderate, and severe. New indentations of the thecal sac of varying severity (mild in 17 patients [38.6%], moderate in 10 [22.7%], and severe in six [13.6%]) had developed at the adjacent segments in 33 (75%) of 44 patients. The degenerative changes were seen at the superior level in 11 patients, inferior level in 10 patients, and at both levels in 12 patients and resulted from both anterior and posterior element degeneration in the majority (23 [69.6%]) of patients. The remote segments showed mild thecal sac indentations in seven patients and moderate indentations in two patients (nine [20.5%] of 44). Compared with the changes at the remote segment, the canal size was significantly decreased at the superior adjacent segment by 0.9 mm (p = 0.007). No patient sustained a new neurological deficit due to adjacent-segment changes. CONCLUSIONS: On short-term follow-up MR imaging, levels adjacent to the fused segment exhibited more pronounced degenerative changes (compared with remote levels) in 75% of patients who had undergone one- or two-level central corpectomy. PMID- 14748567 TI - Laminoplasty for cervical myelopathy caused by subaxial lesions in rheumatoid arthritis. AB - OBJECT: Although controversy exists regarding surgical treatment for rheumatoid subaxial lesions, no detailed studies have been conducted to examine the efficacy of laminoplasty in such cases. To discuss indications for laminoplasty in rheumatoid subaxial lesions, the authors retrospectively investigated clinical and radiological outcomes in patients who underwent laminoplasty for subaxial lesions. METHODS: Thirty patients (11 men and 19 women) underwent laminoplasty for rheumatoid subaxial lesions. The patients were divided into those with mutilating-type rheumatoid arthritis (RA) and those with nonmutilating-type RA according to the number of eroding joints. As of final follow-up examination laminoplasty resulted in improvement of myelopathy in 24 patients (seven with mutilating- and 17 with nonmutilating-type RA) and transient or no improvement in six (five with mutilating- and one with nonmutilating-type RA). In the group with mutilating-type RA, significantly poorer results were displayed (p < 0.05). In most patients preoperative radiographs demonstrated vertebral slippage less than or equal to 5 mm at only one or two levels. Postlaminoplasty deterioration of subaxial subluxation and unfavorable alignment change occurred significantly more often in patients with mutilating-type RA (p < 0.05). CONCLUSIONS: Patients with nonmutilating-type RA can benefit from laminoplasty for myelopathy due to subaxial lesions. PMID- 14748568 TI - Anterior correction of cervical kyphotic deformity: effects on myelopathy, neck pain, and sagittal alignment. AB - OBJECT: Cervical myelopathy may develop as a result of spinal cord compression with or without deformity. The effect of persistent kyphotic deformity on the ability of the cervical cord to recover following decompressive surgery is not known. METHODS: Between 1997 and 2000, a total of 28 patients with progressive myelopathy and kyphotic deformity underwent anterior decompression, deformity correction (0-4 degrees of lordosis), and fusion with anterior plating. Patients received clinical and radiological follow-up care, with independent analysis. Variables assessed included patient characteristics, severity of preoperative myelopathy, neck pain, and cervical sagittal alignment. Twenty-six patients (93%) underwent follow-up review for a minimum of 18 months. Two patients died: one died in the perioperative period and was excluded from further analysis, and in the other only 3 months of follow-up data could be obtained. Local deformity was corrected to neutral or lordosis in 24 cases (89%), and the overall cervical curve was corrected to neutral or lordosis in 20 cases (74%). There was a significant improvement in myelopathy scores in those patients in whom the target (0 to 4 degrees of lordosis) local angle was achieved (p = 0.04). There was a variable change in overall cervical sagittal alignment following local correction. Improvement in myelopathy was unrelated to patient age, previous surgery, or number of segments fused. Improvement in pain score was not related to correction of kyphotic angle. CONCLUSIONS: The correction of sagittal alignment may promote recovery in spinal cord function in patients with kyphotic deformity. PMID- 14748569 TI - Tracheostomy placement in patients with complete cervical spinal cord injuries: American Spinal Injury Association Grade A. AB - OBJECT: The authors sought to identify variables that predispose patients with acute American Spinal Injury Association (ASIA) Grade A cervical spinal cord injury (SCI) to require tracheostomies for ventilator support or airway protection. METHODS: A retrospective analysis was performed of 178 consecutive patients with a cervical ASIA Grade A SCI who were admitted through the Delaware Valley SCI Center at Thomas Jefferson Hospital during a 6-year period. Exclusion criteria included injury occurring more than 48 hours prior to admission, death within 14 days of admission or nontraumatic SCI. Twenty-two patients were excluded based on these criteria. Parameters evaluated in the remaining population (156 patients) included demographics, cervical vertebral ASIA level, tracheostomy placement, pneumonia, premorbid pulmonary disease, smoking history, evidence of direct thoracic/lung trauma, operative intervention, associated appendicular trauma, and preexisting medical comorbidities. The ASIA classification of the 156 patients included in this analysis were C-2 (eight), C 3 (11), C-4 (64), C-5 (36), C-6 (20), C-7 (13), and C-8 (four). Tracheostomies were performed in 107 of these 156 patients. Statistical analysis revealed a significant relationship between tracheostomy and patient age (p = 0.0048), preexisting medical conditions (p = 0.0417), premorbid lung disease (p = 0.0177), higher cervical ASIA level (p < 0.0001), and the presence of pneumonia (p < 0.0001). No patient with a C-8 ASIA A injury required tracheostomy, whereas all C 2 and C-3 ASIA A-injured patients underwent tracheostomies. Patients older than 45 years of age with ASIA A levels between C-4 and C-7 more commonly required tracheostomy (p < 0.005) than patients younger than 45 years of age. CONCLUSIONS: Several risk factors were identified that corresponded to the frequent tracheostomy placement in the acute injury phase after complete cervical SCI. Early tracheostomy may be considered in patients with multiple risk factors to reduce duration of stay in the intensive care unit and facilitate ventilatory weaning. PMID- 14748570 TI - Intravertebral vacuum phenomenon in osteoporotic compression fracture: report of 67 cases with quantitative evaluation of intravertebral instability. AB - OBJECT: The objectives of this study were to: 1) describe the incidence and clinical features of intravertebral vacuum phenomenon (IVVP) in a relatively large number of cases; 2) quantitatively evaluate intravertebral instability and determine the factors affecting instability; and 3) evaluate the efficacy of percutaneous vertebroplasty in the treatment of this phenomenon. METHODS: A retrospective review was conducted of the records of 67 patients with IVVP among 652 consecutive cases of osteoporotic compression fracture. Comparisons between the IVVP group and a control group, a stable group, and an unstable group were conducted. Percutaneous vertebroplasty was performed in all patients. There were 67 patients (10.3%) in whom there were 70 vacuum phenomena of the intravertebral space. Intravertebral vacuum phenomena occurred predominantly in the thoracolumbar junction (81%) and in patients with a longer duration of symptoms (10.6 +/- 9.8 months) compared with the control group. Of 59 vertebrae for which flexion-extension radiographs were available, 26 vertebrae were categorized as stable and 33 as unstable. Twenty-one vertebrae (64%) had undergone compression fracture in the unstable group compared with nine (35%) compression fractures in the stable group. There were 28 (85%) fractures of the wedged vertebrae in the unstable group compared with 16 (61%) fractures in wedged vertebrae in the stable group. Percutaneous vertebroplasty was performed with successful clinical outcome. CONCLUSIONS: Intravertebral vacuum phenomenon is more common than has been previously appreciated. The results of this study indicate that biomechanics, not ischemic or avascular theory, may play an important role in pathogenesis of this phenomenon. Percutaneous vertebroplasty was found to be a minimally invasive and effective procedure for the treatment of IVVP. PMID- 14748572 TI - Experience in the surgical management of spontaneous spinal epidural hematoma. AB - OBJECT: Spontaneous spinal epidural hematoma (SSEH) is a rare disease entity. Although many cases have been reported in the literature, controversy persists as to its origin, diagnosis, and timing of treatment. The authors conducted a study in patients treated in their hospital and report the results. METHODS: Clinical data obtained in 35 patients with SSEH were retrospectively reviewed. Age, sex, history of hypertension, and history of anticoagulation therapy were recorded, and data were analyzed to clarify the possible predisposing factors of SSEH. Neurological outcomes were reappraised using a standardized grading system and correlated with the time interval from initial ictus to surgery, duration of complete neurological deficits, and the rapidity of deterioration of paralysis. Nonparametric methods and Spearman rank-correlation coefficients were used for statistical analysis. CONCLUSIONS: Surgery is a safe and effective procedure to treat SSEH. The disease-related mortality rate was 5.7%, the surgery-related complication rate was 2.9%, and there were no operation-related deaths. Neurological outcome after surgery is positively correlated with preoperative neurological deficits (88.9% complete recovery in patients with incomplete neurological deficits compared with 37.5% in those with complete deficits [p < 0.001]). In patients in whom the time interval from initial ictus was shorter (< 48 hours) and in whom the duration of complete neurological symptoms was also briefer (< 12 hours), there is a positive correlation with better neurological and functional recovery (p < 0.05). PMID- 14748571 TI - Microsurgical interlaminary vertebro- and kyphoplasty for severe osteoporotic fractures. AB - OBJECT: Percutaneous vertebro- and kyphoplasty have become established methods for the treatment of uncomplicated osteoporotic vertebral fractures. In the setting of severe fractures involving fragmentation of the posterior wall and neural compromise, however, decompressive surgery cannot be performed and epidural cement leakage is poorly controlled. A microsurgical interlaminary approach for vertebro- and kyphoplasty was developed to allow spinal decompression and control of the spinal canal during augmentation. METHODS: Interlaminary vertebro- or kyphoplasty was performed in 24 patients with osteoporotic fractures involving neural compression or posterior wall fragmentation. After unilateral microsurgical fenestration, decompression of the spine, and gentle mobilization of the thecal sac, vertebro- or kyphoplasty was performed directly through the posterior wall of the fractured vertebral body. Cement was injected under microscopic and fluoroscopic control, with the option of immediate exploration of the exposed spinal canal. Thirty-four levels (T-8 to L-5) were treated. Mean blood loss was less than 100 ml and augmentation added 10 to 40 minutes to the entire procedure. Cement leakage associated with the kyphoplasty procedure was less than that in vertebroplasty. There were no major complications. One patient was lost to follow up. Clinical outcome was good or excellent in 17 of the 23 patients available for follow-up (1 to 31-month) evaluation. CONCLUSIONS: The present microsurgical interlaminary approach for vertebro- and kyphoplasty enables treatment of severe osteoporotic fractures involving fragmentation of the posterior wall and neural compromise. Decompressive surgery is possible and the risk of epidural cement leakage is controlled intraoperatively. This technique can be regarded as a procedure on the treatment continuum between percutaneous augmentation and conventional open reconstruction. PMID- 14748573 TI - Rotational and transpositional flaps for the treatment of spinal wound dehiscence and infections in patient populations with degenerative and oncological disease. AB - OBJECT: Wound-related complications following complex posterior spine procedures may result in the need for serial debridements and may place the instrumentation at risk. Numerous treatments have been advocated for this problem, but each has limitations. In this article the authors discuss the experience from two large teaching institutions at which rotational and transpositional flaps were used in the management of deep wound infections and dehiscences. METHODS: The authors generated a list of patients treated via posterior or posterolateral approaches for metastatic tumors or complex degenerative disorders in whom wound complications subsequently developed. Data were obtained from the medical records and reviewed retrospectively. Thirty-seven patients were treated with rotational or transpositional flaps at the two institutions during the study period. Patients underwent a mean of 1.3 procedures for the treatment of wound healing problems, and cultures were positive in 70%. In three patients (8%) this treatment failed due to protrusion of hardware through the skin or repeated dehiscence requiring reclosure. Spinal instrumentation was salvaged in 97% of the cases. CONCLUSIONS: The use of local tissue flaps is advantageous for treatment of posterior wound complications due to spine surgery. In this procedure highly vascularized tissue is used to increase healing, accelerate clearance of bacteria, and fill any dead space. PMID- 14748574 TI - Mitomycin C in preventing spinal epidural fibrosis in a laminectomy model in rats. AB - OBJECT: Extensive epidural fibrosis after lumbar surgery may be the underlying cause in most cases of failed-back surgery syndrome. Various materials have been used to prevent epidural fibrosis, but only moderate success has been shown. Mitomycin C, an alkylosing antibiotic substance isolated from Streptomyces caespitosus, potentially supresses fibroblast proliferation after surgery. In this study, the authors investigated the effect of mitomycin C by local application on spinal epidural fibrosis in a rat laminectomy model. METHODS: Five Wistar rats underwent laminectomy at cervical, thoracic, and lumbar levels. Based on data obtained from ophthalmological studies, mitomycin C was applied to the laminectomy sites in various concentrations (0.01, 0.05, and 0.1 mg/ml). One laminectomy site in each rat was left untreated and thus served as a control. Evoked potentials were measured pre- and postoperatively, and all rats underwent clinical evaluation. Mobility status and evidence of neurological deficit were recorded. Twelve weeks later, the rats were killed, and the spinal column, including surrounding muscle tissue, was removed en bloc, decalcified, and fixed in formaldehyde. Epidural fibrosis was evaluated histologically. In all mitomycin C-treated laminectomy sites, epidural scarring was significantly reduced compared with control sites. Remarkably, dural adhesions were absent in laminectomy defects treated with mitomycin C concentrations of 0.05 and 0.1 mg/ml. Moderate to marked epidural fibrosis with adhesion to the dura mater was noted at sites receiving 0.01 mg/ml of mitomycin C. All control sites showed dense epidural fibrosis with marked dura adherence. CONCLUSIONS: In this experimental model, mitomycin C applied locally at a concentration of 0.1 mg/ml effectively reduced epidural fibrosis, completely avoided dural adherence, and induced no side effects. PMID- 14748576 TI - Transvertebral interbody cage and pedicle screw fixation for high-grade spondylolisthesis. Case report. AB - Surgical stabilization of high-grade lumbosacral spondylolisthesis is clinically challenging, and the success of deformity reduction and fusion varies. The authors describe a patient with Grade III spondylolisthesis at L5-S1. Partial reduction was achieved and fusion involved pedicle screw fixation and a posterior transvertebral interbody cage. This patient had developed progressive spondylolisthesis after decompression and posterolateral fusion for L5-S1 spondylolisthesis failed. Clinical and early radiographic results were excellent. Transsacral cage fixation can be considered a viable option to buttress the region in which high-grade L5-S1 spondylolisthesis has been reduced. The cage provides substrate for interbody arthrodesis and acts as a biomechanical stabilizer that helps prevent pedicle screw failure. PMID- 14748575 TI - Bladder function after incomplete spinal cord injury in mice: quantifiable outcomes associated with bladder function and efficiency of dehydroepiandrosterone as a therapeutic adjunct. AB - OBJECT: The authors conducted a study to establish outcomes associated with bladder function in a mouse model of spinal cord injury (SCI) and to assess the sensitivity of these outcomes in determining the efficacy of pharmacological treatments. METHODS: A mouse model of moderate contusive SCI was used. Outcome parameters included physiological, behavioral, and morphological measurements. To test the sensitivity of these outcomes, the authors used a dehydroepiandrosterone (DHEA) treatment that they had previously shown to promote neurological recovery effectively after SCI. A behavioral scale was used to identify the day at which autonomic function of the bladder was recovered. The reduction in the daily volume of urine during the period of functional recovery paralleled this scale. They then determined the day postinjury at which the functional differences between the vehicle- and DHEA-treated mice exhibited the maximal amplitude. Changes were measured in the composition of the extracellular matrix relative to collagen expression in the layer muscularis of the detrusor at this time point. They found that SCI increases the ratio of collagen type III to collagen type I in the detrusor. Moreover, in the DHEA-treated group, this ratio was similar to that demonstrated in sham-operated mice, establishing the sensitivity of this outcome to assess therapeutic benefits to the bladder function. They next examined the relationship between measurements of neurological recovery and controlled voiding by using cluster analysis. CONCLUSIONS: The authors found that early recovery of controlled voiding is predictive of motor recovery. PMID- 14748577 TI - Improved repair of cervical esophageal fistula complicating anterior spinal fusion: free omental flap compared with pectoralis major flap. Report of four cases. AB - Esophageal injury is a serious complication of anterior cervical fusion. A team approach to the management of these cases is described. The authors performed spinal assessment, control of the fistula, and interposition of a vascularized flap between the spine and the esophagus. They compared the overall efficacy of the pectoralis major flap (pedicled; two cases) and omental flap (free; two cases). PMID- 14748578 TI - Thoracic paraplegia due to missed thoracic compressive lesions after lumbar spinal decompression surgery. Report of three cases. AB - The authors discuss the cases of three patients in whom thoracic paraplegia developed after lumbar spinal decompressive surgery for slight lumbar spinal canal stenosis. Careful computerized tomography myelography and magnetic resonance imaging examination of the thoracic spine revealed another compressive lesion (spinal cord tumor, disc herniation, osteophyte of vertebral body, and ossification of the ligamentum flavum). Additional thoracic decompressive surgery provided partial amelioration of each patient's neurological condition. The authors suggest that to avoid such a complication physical and radiographic examination of the thoracic spine should be performed preoperatively if the lumbar imaging is inconclusive. PMID- 14748579 TI - Giant cell ependymoma of the spinal cord. Case report and review of the literature. AB - Several rare histological variants of ependymoma have been described. The authors report on a patient in whom cervical spinal cord astrocytoma was originally diagnosed after evaluation of a limited biopsy specimen. More abundant tissue obtained during gross-total resection included areas of well-differentiated ependymoma. The histological features of the tumor were extremely unusual, with a major component of pleomorphic giant cells. Its histological, immunohistochemical, and electron microscopic features, however, were consistent with ependymoma. Only two cases of terminal filum and two of supratentorial giant cell variant of ependymoma have been reported. To the authors' knowledge, this represents the first case of giant cell ependymoma of the spinal cord. The clinical significance is the potential for misdiagnosis with anaplastic (gemistocytic) astrocytoma, especially in cases in whom limited biopsy samples have been obtained. PMID- 14748580 TI - Noncovalent self-assembly of bicyclo[4.2.2]diketopiperazines: influence of saturation in the bridging carbacyclic ring. AB - [structure: see text] We have prepared 7,9-diazabicyclo[4.2.2]dec-3-ene-8,10 dione (3) and 7,9-diazabicyclo[4.2.2]decane-8,10-dione (4), which differ by virtue of the degree of unsaturation in the bridging carbacyclic tether on a 2,5 diketopiperazine. Remarkably different self-assembly patterns were observed in the solid state for the two compounds, attributed to subtle variations in the conformational constraints imposed by the tether. PMID- 14748581 TI - Synthesis and first molecular structure of a bis-2-spiro-1-boraadamantane derivative. AB - [reaction: see text] A new method for synthesizing the 2-spiro-boraadamantane pyridine complex (2) from 1-ethynylcyclohexylmethyl ether has been developed. The chemistry has been applied to the synthesis of bis-2-spiro-1 boraadamantane.pyridine (1) from trans-1,4-diethynyl-1,4-dimethoxycyclohexane (8). This bis-Lewis acid serves as a self-assembling molecular building block with difunctional Lewis bases. PMID- 14748582 TI - Synthesis of the putative structure of eupomatilone-6. AB - [reaction: see text] The synthesis of eupomatilone-6 (1) has been achieved by using Suzuki coupling, Sharpless asymmetric dihydroxylation, and intramolecular Horner-Wadsworth-Emmons reactions. The spectroscopic studies carried out on synthetic eupomatilone-6 do not agree with those reported for the natural product, and therefore revision of the assigned structure is warranted. PMID- 14748583 TI - Water-soluble camptothecin derivatives that are intrinsic topoisomerase I poisons. AB - [structure: see text] In an effort to improve the water solubility of camptothecin, four 20-O-phosphate and phosphonate analogues have been prepared. These analogues are freely water soluble, stable at physiological pH, and stabilize the human topoisomerase I-DNA covalent binary complex with the same sequence selectivity as camptothecin itself. All four compounds inhibited the growth of yeast expressing human topoisomerase I in an enzyme-dependent fashion. PMID- 14748584 TI - Remarkable effect of a silicon group on the stereoselectivity of radical 5-exo trig cyclizations. AB - [reaction: see text] Sulfonyl radical mediated 5-exo-trig cyclization of chiral 3 silylhepta-1,6-dienes has been shown to provide cyclopentanes having up to four stereogenic centers with an unexpectedly high level of stereocontrol. PMID- 14748585 TI - Total synthesis of carbazoquinocin C: application of the o-benzannulation of Fischer carbene complexes to carbazole-3,4-quinone alkaloids. AB - [reaction: see text] The photoinduced o-benzannulation of 3-(2 vinyl)indolylcarbene complexes provides a direct route to carbazole derivatives that are oxygenated in the 3- and 4-positions. This reaction is quite efficient and provides for a unique synthesis of the lipid peroxidation inhibitor carbazoquinocin C. PMID- 14748586 TI - Quinoxaline excision: a novel approach to tri- and diquinoxaline cavitands. AB - [structure: see text] Selective excision of one or two quinoxaline units from tetraquinoxaline cavitand using catechol and base in DMF yields tri- and diquinoxaline cavitands in yields of up to 71%. PMID- 14748588 TI - Diastereoselective reduction of cyclopropenylcarbinol: new access to anti cyclopropylcarbinol derivatives. AB - [reaction: see text] Cyclopropenylcarbinol derivatives are regio- and diastereoselectively reduced with LiAlH(4) in Et(2)O into trans cyclopropylcarbinols as a single isomer. The regioselectivity of the hydroalumination reaction on the cyclopropenyl ring has been mapped out by deuterium labeling experiments. PMID- 14748587 TI - Citrafungins A and B, two new fungal metabolite inhibitors of GGTase I with antifungal activity. AB - [structure: see text] Screening of natural products extracts led to the discovery of citrafungins A and B, two new fungal metabolites of the alkylcitrate family that are inhibitors of GGTase I of various pathogenic fungal species with IC(50) values of 2.5-15 microM. These compounds exhibited antifungal activities with MIC values of 0.40-55 microM. The isolation, structure elucidation, relative and absolute stereochemistry, and biological activities of citrafungins are described. PMID- 14748589 TI - Direct vinylogous Mannich-type reactions via ring opening and rearrangement of vinyloxiranes. AB - [reaction: see text] The first synthetic application of 2-methyl-2-vinyloxirane as a masked dienolate has been successfully demonstrated in the direct vinylogous Mannich-type reaction with an alpha-imino ester as an electrophile. The Mannich adduct was a useful intermediate en route to cis-5-substituted pipecolinic acid ethyl ester under simple hydrogenation conditions. PMID- 14748591 TI - A novel approach to 2-arylated quinolines: electrocyclization of alkynyl imines via vinylidene complexes. AB - [reaction: see text] Alkynyl imines underwent [4 + 2] electrocyclization in the presence of 20 mol % W(CO)(5)(THF) to give 2-arylated quinolines in good yields. A deuterium labeling study suggests that the reaction proceeds via a tungsten vinylidene complex. PMID- 14748590 TI - Synthesis of site-specifically labeled arachidonic acids as mechanistic probes for prostaglandin H synthase. AB - [structure: see text] Prostaglandin H synthase catalyzes the first committed step in the biosynthesis of prostaglandins and thromboxane. Herein we report the synthesis of four site-specifically labeled arachidonic acids for investigation of the radical intermediate formed during this enzymatic reaction. Two compounds were prepared using a common C9-C11 fragment, while another target was synthesized using a previously reported advanced intermediate. An alkyne coupling followed by hydrogenation and Wittig reaction was used to prepare the final labeled substrate. PMID- 14748592 TI - Palladium-catalyzed cross-coupling of stereospecific potassium cyclopropyl trifluoroborates with aryl bromides. AB - [reaction: see text] Stereospecific cyclopropanation of alkenylboronic esters of pinacol followed by in situ treatment with excess KHF(2) afforded the corresponding potassium cyclopropyl trifluoroborates in high yields, which then underwent Suzuki-Miyaura cross-coupling reactions with aryl bromides to give cyclopropyl-substituted arenes in good yields with retention of configuration. This promises to be a useful method for the synthesis of enantiomerically pure cyclopropanes. PMID- 14748593 TI - Preparation of beta-cyclodextrin polyrotaxane: photodimerization of pseudo polyrotaxane with 2-anthryl and triphenylmethyl groups at the ends of poly(propylene glycol). AB - [structure: see text] A polyrotaxane containing beta-cyclodextrins has been prepared by photoreactions of a precursor complex between beta-cyclodextrin with poly(propylene glycol) having a triphenylmethyl group at one end and a 2-anthryl group at the other end. PMID- 14748594 TI - First total synthesis of mycothiol and mycothiol disulfide. AB - [reaction: see text] The first total synthesis of mycothiol and mycothiol disulfide was achieved by linking D-2,3,4,5,6-penta-O-acetyl-myo-inositol, O (3,4,6-tri-O-acetyl)-2-azido-2-deoxy-alpha,beta-D-glucopyranosyl) trichloroacetimidate, and N,S-diacetyl-L-cysteine and deprotecting peracetylated mycothiol. The first full spectral characterization is reported for underivatized mycothiol. The structure of mycothiol was confirmed by spectral analysis of the known bimane derivative. PMID- 14748595 TI - Small cavitands specifically binding a water molecule. AB - [structure: see text] Three new C(2v) cavitands based on resorcin[4]arene bind water specifically at low temperature in CD(2)Cl(2) or CDCl(3) due to their complementarity to water as well as the solvophobic interaction. The averaged DeltaH(o) and DeltaS(o) values are -2.3 kcal mol(-1) and -128 cal mol(-1) K(-1), which gave the averaged -DeltaG(o) of 1.9 kcal mol(-1) at -50 degrees C in water saturated CD(2)Cl(2). PMID- 14748596 TI - A readily synthesized and highly active epoxide carbonylation catalyst based on a chromium porphyrin framework: expanding the range of available beta-lactones. AB - [reaction: see text] Catalytic carbonylation of epoxides to beta-lactones was effected by a highly active and selective bimetallic catalyst comprised of a chromium(III) porphyrin cation and a cobalt tetracarbonyl anion. The complex is readily synthesized from commercially available compounds in high yield. Carbonylation of numerous linear epoxides, as well as bicyclic epoxides derived from 8- and 12-membered hydrocarbons, proceeded with high activity, selectivity, and yield. PMID- 14748597 TI - Transition-metal-catalyzed formation of trans alkenes via coupling of aldehydes. AB - [reaction: see text] Rh(2)(OAc)(4) catalyzed the formation of exclusively trans fluorinated alkenes from aldehydes and pentafluorobenzaldehyde tosylhydrazone salts, which were readily prepared from pentafluorobenzaldehyde using the Bamford Stevens reaction. A series of pentafluorophenyl-containing alkenes were synthesized from aldehydes in moderate to good yields under mild reaction conditions in a one-pot reaction. It is the first report of coupling two different aldehydes to form exclusively trans alkenes. PMID- 14748598 TI - First carbamates conversion to amides by simple alkyl group transfer from trialkylalanes. AB - [reaction: see text] N-Monosubstituted carbamates are cleanly converted to amides under treatment with trialkylaluminum. This reaction involves an aluminum assisted internal delivery of alkyl groups. It can be applied to new and mild protecting group strategies for alcohols. PMID- 14748599 TI - Model systems for flavoenzyme activity: interplay of hydrogen bonding and aromatic stacking in cofactor redox modulation. AB - [structure: see text] A model system has been developed to study the synergy between aromatic stacking and hydrogen bonding in the binding of a flavin derivative. The results show that the identity of both the hydrogen bonding and pi-stacking units strongly determine the overall receptor affinity for flavin in both the oxidized and radical anion forms. PMID- 14748600 TI - Microwave-assisted Paal-Knorr reaction. A rapid approach to substituted pyrroles and furans. AB - [reaction: see text] An array of tetrasubstituted pyrroles (and trisubstituted furans) was obtained using a simple three-step procedure. Functional homologation of a beta-ketoester with an aldehyde followed by oxidation gave a series of differently substituted 1,4-dicarbonyl compounds that can be rapidly cyclized with the Paal-Knorr procedure carried out under microwave irradiation. PMID- 14748601 TI - Anion radicals of di-trans-[12]annulene and heptalene in a one-pot synthesis from a common fire retardant. AB - [reaction: see text] Low temperature (-100 degrees C) dehydrohalogenation of 1,2,5,6,9,10-hexabromocyclododecane (a common fire retardant) with potassium tert butoxide in THF followed by one-electron reduction yields the anion radical of the di-trans form of [12]annulene. This system yields a well-resolved EPR signal that reveals that most of the spin density resides on one side (the planar side) of the anion radical. Five of the carbons in this [12]annulene system are twisted from the plane of the remaining seven carbons, and the rate of rearrangement between the degenerate conformations is on the EPR time scale (k = 10(6)-10(7) s( 1)). Warming of the solution results in the formation of a sigma-bond between the two internal carbons, loss of molecular hydrogen, and consequent generation of the anion radical of heptalene. Tractable quantities of neutral heptalene can be obtained via the reoxidation of this anion radical with iodine. PMID- 14748602 TI - Synthesis of ether oligomers. AB - [structure: see text] Hydroxyaromatic aldehydes and ketones were used as building blocks to prepare ether oligomers. An iterative two-step protocol involving Mitsunobu coupling and carbonyl reduction provided a protecting-group-free route with high yields. Activity screening of an 84-member library against proteases led to the discovery of micromolar inhibitors for trypsin, chymotrypsin, and subtilisin. PMID- 14748603 TI - Time-resolved IR detection and study of an iminooxirane intermediate. AB - [reaction: see text] Iminooxirane 4 has been generated by the reaction of phenylchlorocarbene (2) with phenyl isocyanate (3) and studied by nanosecond time resolved infrared spectroscopy. Iminooxirane 4 (1830 cm(-1)) isomerizes to alpha lactam 5 (1880 cm(-1)) at an observed rate of 3 x 10(4) s(-1). Peak assignments were confirmed by a combination of B3LYP calculations and isotopic labeling. An acyclic transition state was found computationally for the isomerization reaction. PMID- 14748604 TI - Alpha,alpha-disubstituted boron enolates in the asymmetric synthesis of quaternary carbon centers. AB - [reaction: see text] Reduction of alpha,alpha-disubstituted thioglycolate amides with lithium di-tert-butylbiphenylide affords alpha,alpha-disubstituted enolates with high Z/E selectivity. Transmetalation of the enolates with dicyclohexylboron bromide facilitates highly diastereoselective aldol reaction with aromatic and alpha,beta-unsaturated aldehydes. PMID- 14748605 TI - Enantiocontrolled synthesis of jasmonates via tandem retro-Diels-Alder-ene reaction activated by a silyl substituent. AB - [reaction: see text] An enantiocontrolled synthesis of (-)-methyl 6-epi-cucurbate and (+)-methyl jasmonate was established from a chiral tricyclic lactone via a new type of tandem retro-Diels-Alder-ene reaction activated by a trimethysilyl substituent as the key step. PMID- 14748607 TI - Cyclizations of N-stannylaminyl radicals onto nitriles. AB - [reaction: see text] Stannylaminyl radicals derived from radical reactions of Bu(3)SnH with azidoalkylmalononitriles exhibit highly efficient 5- and 6-exo cyclization onto either nitrile group to give aminoiminyl radicals that in turn are reduced to amidines or undergo successive 5-exo cyclization onto an internal alkene. PMID- 14748606 TI - Total synthesis of aigialomycin D. AB - [reaction: see text] The first total synthesis of resorcinylic macrolide aigialomycin D was described. The resorcinylic moiety was constructed by a highly efficient Diels-Alder reaction using a disiloxydiene and a 14-membered "ynolide" as the dienophile synthesized by ring-forming olefin metathesis. PMID- 14748608 TI - Direct electrophilic silylation of terminal alkynes. AB - [reaction: see text] A variety of alkynylsilanes were efficiently prepared via direct silylation of terminal alkynes with aminosilanes in the presence of zinc halides. Base- and nucleophile-sensitive functionalities were perfectly tolerated under the above reaction conditions. Initial mechanistic studies support the electrophilic character of this transformation. PMID- 14748609 TI - Preparation and coordination properties including catalytic activities of a bulky 2-methyl-3-thioxo-1,3-diphosphapropene. AB - [reaction: see text] 2-Methyl-3,3-diphenyl-3-thioxo-1-(2,4,6-tri-tert butylphenyl)-1,3-diphosphapropene, which bears a P=C-P=S skeleton, was prepared and used as a ligand of transition-metal complexes. The palladium complexes containing the ligated 3-thioxo-1,3-diphophapropene were stable and used for catalytic reactions such as cross-couplings and direct conversion of allyl alcohol to allylaniline. PMID- 14748610 TI - Asymmetric total synthesis of bacillariolide III, a marine oxylipin. AB - [reaction: see text] The asymmetric total synthesis of bacillariolide III has been achieved via 15 linear steps in 14.6% overall yield. The key feature of this synthetic route involves the highly stereoselective construction of the vinyl substituted bicyclic lactone by an intramolecular Pd(0)-catalyzed allylic alkylation and its facile conversion to the hydroxy bicyclic lactone skeleton of bacillariolide III, induced by stereoselective vinylcerium addition to the aldehyde. In addition, the (Z)-pentenoic acid was efficiently introduced by the internal hydroxy group tethered ring-closing metathesis (RCM). PMID- 14748611 TI - Heck-type arylation of 2-cycloalken-1-ones with arylpalladium intermediates formed by decarboxylative palladation and by aryl iodide insertion. AB - [reaction: see text] A palladium-catalyzed decarboxylative arylation reaction was shown to produce Heck-type coupling products using a number of different arene carboxylic acid and 2-cycloalken-1-one substrates. The more conventional Heck coupling of an aryl iodide and a 2-cycloalken-1-one reactant was also briefly explored for comparison, where it was found that phosphine-free (Jeffery) conditions afforded the highest yield of product. PMID- 14748612 TI - Chloromethanesulfonylethene and dichloromethanesulfonylethene: new reagents for tandem Diels-Alder/Ramberg-Backlund reactions. AB - [reaction: see text] Chloromethanesulfonylethene (3a) and dichloromethanesulfonylethene (3b) were prepared by oxidation of the adducts of ethylene and ClCH(2)SCl or Cl(2)CHSCl, respectively, followed by NaHCO(3) dehydrochlorination. With dienes, 3a gave Diels-Alder adducts that, with base, underwent Ramberg-Backlund reaction, giving products corresponding to the adducts of the dienes and allene. Similarly, 3b gave Diels-Alder adducts that, with base in the presence of the novel chlorine source MeSO(2)CCl(3), cleanly afforded products corresponding to the adducts of the dienes and 1,1-dichloropropa-1,2 diene. PMID- 14748613 TI - Poly(ethylene glycol)-supported TEMPO: an efficient, recoverable metal-free catalyst for the selective oxidation of alcohols. AB - [structure: see text] Poly(ethylene glycol)-supported TEMPO (PEG-TEMPO) has been prepared, and its catalytic activity in the chemoselective oxidation of alcohols with stoichiometric amounts of organic or inorganic oxidants has been investigated. The new metal-free catalyst exhibits high activity and is easily removed from the reaction mixture by filtration. Recycling experiments showed that PEG-TEMPO can be reused up to six times with no loss of catalytic activity. PMID- 14748614 TI - Diastereoselective additions of chiral vinylzinc reagents to alpha-chiral aldehydes. AB - [reaction: see text] Additions of vinylic zinc bromide reagents to alpha-chiral aldehydes (R(1) = CH(2)OTBS, R(2) = Me; R(1) = Me, R(2) = OTBS) in the presence of lithiated (+)- or (-)-N-methylephedrine proceed with predominant reagent control to afford anti or syn adducts stereoselectively, except when the aldehydes possess an alkoxy substituent at the alpha- or beta-positions (R(1) = Me, R(2) = OBn; R(1) = CH(2)OBn, R(2) = Me), in which case chelation-controlled adducts predominate. PMID- 14748615 TI - Synthesis of beta-substituted alpha-amino acids via Lewis acid promoted radical conjugate additions to alpha,beta-unsaturated alpha-nitro esters and amides. AB - [reaction: see text] Beta-substituted alpha,beta-unsaturated alpha-nitro esters and amides undergo radical conjugate additions when treated with an appropriate Lewis acid. Deuterium studies revealed that the acidic alpha-stereocenter of the alpha-nitro ester products does not racemize under strictly controlled workup conditions. The alpha-nitro amides did racemize significantly during chromatography, but this could be greatly minimized by subjecting the crude adducts to subsequent transformations. The conjugate addition products can be elaborated into beta-substituted alpha-amino acids in two simple steps. PMID- 14748616 TI - Carbonylation of lithium enolates with carbon monoxide mediated by selenium. AB - [reaction: see text] Lithium enolates of ketones and aldehydes undergo carbonylation with carbon monoxide with the aid of selenium under mild conditions to yield beta-keto and beta-formyl selenol esters after trapping with alkyl iodides. This reaction proceeds via a unique carbonylation mechanism comprised of O-carbonylation and subsequent migration of the SeCO moiety to the alpha-carbon. PMID- 14748617 TI - Antiepileptic-induced resistance to neuromuscular blockers: mechanisms and clinical significance. AB - Prolonged administration of antiepileptic drugs is associated with several drug interactions. In the field of anaesthesia and critical care, patients exhibit both sensitivity and resistance to non-depolarising neuromuscular blockers (NDNMBs) after acute and long-term administration of antiepileptic drugs, respectively. Although antiepileptic therapy alone has only mild neuromuscular effects, acutely administered antiepileptic drugs can potentiate the neuromuscular effects of NDNMBs as a result of direct pre- and post-junctional effects. Resistance to NDNMBs during long-term antiepileptic therapy is due to multiple factors operating alone or in combination, including induction of hepatic drug metabolism, increased protein binding of the NDNMBs and/or upregulation of acetylcholine receptors. PMID- 14748618 TI - Clinical pharmacokinetics of everolimus. AB - Everolimus is an immunosuppressive macrolide bearing a stable 2-hydroxyethyl chain substitution at position 40 on the sirolimus (rapamycin) structure. Everolimus, which has greater polarity than sirolimus, was developed in an attempt to improve the pharmacokinetic characteristics of sirolimus, particularly to increase its oral bioavailability. Everolimus has a mechanism of action similar to that of sirolimus. It blocks growth-driven transduction signals in the T-cell response to alloantigen and thus acts at a later stage than the calcineurin inhibitors ciclosporin and tacrolimus. Everolimus and ciclosporin show synergism in immunosuppression both in vitro and in vivo and therefore the drugs are intended to be given in combination after solid organ transplantation. The synergistic effect allows a dosage reduction that decreases adverse effects. For the quantification of the pharmacokinetics of everolimus, nine different assays using high performance liquid chromatography coupled to an electrospray mass spectrometer, and one enzyme-linked immunosorbent assay, have been developed. Oral everolimus is absorbed rapidly, and reaches peak concentration after 1.3-1.8 hours. Steady state is reached within 7 days, and steady-state peak and trough concentrations, and area under the concentration-time curve (AUC), are proportional to dosage. In adults, everolimus pharmacokinetic characteristics do not differ according to age, weight or sex, but bodyweight-adjusted dosages are necessary in children. The interindividual pharmacokinetic variability of everolimus can be explained by different activities of the drug efflux pump P glycoprotein and of metabolism by cytochrome P450 (CYP) 3A4, 3A5 and 2C8. The critical role of the CYP3A4 system for everolimus biotransformation leads to drug drug interactions with other drugs metabolised by this cytochrome system. In patients with hepatic impairment, the apparent clearance of everolimus is significantly lower than in healthy volunteers, and therefore the dosage of everolimus should be reduced by half in these patients. The advantage of everolimus seems to be its lower nephrotoxicity in comparison with the standard immunosuppressants ciclosporin and tacrolimus. Observed adverse effects with everolimus include hypertriglyceridaemia, hypercholesterolaemia, opportunistic infections, thrombocytopenia and leucocytopenia. Because of the variable oral bioavailability and narrow therapeutic index of everolimus, blood concentration monitoring seems to be important. The excellent correlation between steady-state trough concentration and AUC makes the former a simple and reliable index for monitoring everolimus exposure. The target trough concentration of everolimus should range between 3 and 15 microg/L in combination therapy with ciclosporin (trough concentration 100-300 microg/L) and prednisone. PMID- 14748619 TI - Clinical pharmacokinetics of nateglinide: a rapidly-absorbed, short-acting insulinotropic agent. AB - The prevalence and medical and economic impact of type 2 diabetes mellitus is increasing in Western societies. New agents have been developed that act primarily to reduce postprandial glucose excursions, which may be of particular significance now that postprandial glucose excursions are known to be correlated with cardiovascular morbidity and mortality. Nateglinide is a phenylalanine derivative that blocks K+ channels in pancreatic beta-cells, facilitating insulin secretion. Nateglinide sensitises beta-cells to ambient glucose, reducing the glucose concentration needed to stimulate insulin secretion. The pharmacokinetics of nateglinide are characterised by rapid absorption and elimination, with good (73%) bioavailability. Nateglinide is more rapidly absorbed when given 0-30 minutes prior to meal ingestion than if given during the meal. Nateglinide is extensively metabolised, primarily by cytochrome P450 2C9, and eliminated primarily by the kidney. Nateglinide pharmacokinetics are linear over the dose range 60-240 mg. No significant pharmacokinetic alterations occur in renally impaired patients, in the elderly, or in mildly hepatically impaired patients. Nateglinide administered prior to meals stimulates rapid, short-lived insulin secretion in a dose-dependent manner, thus decreasing mealtime plasma glucose excursions. Its effects on insulin secretion are synergistic with those of a meal. With increasing nateglinide doses, the risk of hypoglycaemia also increases, but its incidence is low. Even if a meal is missed, and the patient skips the dose of nateglinide (as recommended in the event of a missed meal), the incidence of subsequent hypoglycaemia remains low compared with long-acting agents. The postprandial insulinotropic effects of nateglinide are more rapid than those of repaglinide and more rapid and greater than those of glibenclamide (glyburide), while producing less prolonged insulin exposure and less risk of delayed hypoglycaemia. Further investigation is required to determine if nateglinide inhibition of postprandial glucose excursions will help to prevent diabetic complications or preserve pancreatic beta-cell function. PMID- 14748620 TI - Pharmacokinetics and safety of ebastine in patients with impaired hepatic function compared with healthy volunteers: a phase I open-label study. AB - OBJECTIVE: To assess the differences between patients with hepatic insufficiency and healthy subjects with regard to the pharmacokinetics, cardiac safety and overall safety of ebastine and its active metabolite carebastine. DESIGN: Open label parallel-group study. PARTICIPANTS: 24 patients with varying degrees of hepatic insufficiency, as categorised by the Child-Pugh classification, and 12 healthy volunteers. METHODS: Healthy subjects and patients with Child-Pugh class A (n = 8) or B (n = 8) received ebastine 20 mg once daily for 7 days. Patients with Child-Pugh class C (n = 8) [single or repeated dose] received ebastine 10 mg. Plasma concentrations of ebastine and carebastine were determined for 23.5 hours following the initial dose on day 1 and for 96 hours following the dose on day 7 by using a sensitive liquid chromatography-tandem mass spectrometry assay with a minimum quantifiable limit of 0.05 microg/L for ebastine and 1.00 microg/L for carebastine. Hepatic function was assessed by blood clearance of indocyanine green 0.5 mg/kg administered intravenously on day 2. Cardiac and overall safety parameters were monitored. RESULTS: Overall, the pharmacokinetics of ebastine were not modified by hepatic impairment. No correlation between ebastine pharmacokinetics and hepatic function, as expressed by indocyanine green clearance, was observed. Comparison of the effective half-life of ebastine and carebastine between groups did not show relevant differences. Therefore, no apparent accumulation of ebastine occurred, and steady-state concentrations of ebastine and carebastine were predictable from single-dose pharmacokinetics both in healthy subjects and in hepatically impaired patients. Finally, no apparent difference was noted in the safety of ebastine between patients with hepatic insufficiency and healthy subjects as assessed by evaluation of adverse events, vital signs and laboratory parameters. CONCLUSION: Ebastine can be safely administered to patients with impaired hepatic function, as no clinically important differences can be anticipated from the pharmacokinetics and safety profile of ebastine/carebastine as compared with healthy subjects. Nevertheless, the dosage used in severely impaired patients (10mg daily) was half that used in patients with mild to moderate impairment, and any comedication did not include drugs affecting liver function; in clinical practice, both these factors should be taken into account. PMID- 14748623 TI - Endovascular techniques in the management of acute arterioenteric fistulas. AB - PURPOSE: To report the use of endovascular techniques to emergently treat hemorrhagic complications of rare arterioenteric fistulas. CASE REPORTS: Two patients, a 71-year-old man and a 61-year-old woman, presented with acute bleeding arising from primary arterioenteric fistulas. In the first patient, a fistula between the iliac artery and the small intestine complicated laparoscopic treatment of acute appendicitis. In the second patient, irradiation of a metastatic cervical carcinoma led to a fistula between the right iliac artery and the terminal ileum. In both patients, the hemorrhage was controlled with implantation of a Jostent Peripheral Stent-Graft. The man is alive at 3 years with a patent endograft, but the woman died 1 month after treatment from complications of tumor progression. CONCLUSIONS: Endovascular application of covered stents provides an alternative treatment, avoiding extensive surgery. In cases of neoplastic erosion of a large vessel, endovascular stenting can offer a palliative solution. PMID- 14748621 TI - Population pharmacokinetics and pharmacodynamics of mitomycin during intraoperative hyperthermic intraperitoneal chemotherapy. AB - BACKGROUND: During recent years, cytoreductive surgery combined with hyperthermic intraperitoneal chemotherapy (HIPEC) with mitomycin has been used for various malignancies. OBJECTIVE: To characterise the population pharmacokinetics and pharmacodynamics of mitomycin during HIPEC. METHODS: Forty-seven patients received mitomycin 35 mg/m2 intraperitoneally as a perfusion over 90 minutes. Mitomycin concentrations were determined in both the peritoneal perfusate and plasma. The observed concentration-time profiles were used to develop a population pharmacokinetic model using nonlinear mixed-effect modelling (NONMEM). The area under the plasma concentration-time curve (AUC) was related to the haematological toxicity. RESULTS: Concentration-time profiles of mitomycin in perfusate and plasma were adequately described with one- and two-compartment models, respectively. The average volume of distribution of the perfusate compartment (V1) and rate constant from the perfusate to the systemic circulation (k12) were 4.5 +/- 1.1L and 0.014 +/- 0.003 min(-1), respectively (mean +/- SD, n = 47). The average volume of distribution of the central plasma compartment (V2), clearance from the central compartment (CL) and volume of distribution of the peripheral plasma compartment (V3) were 28 +/- 16L, 0.55 +/- 0.18 L/min and 36 +/ 8L, respectively. The relationship between the AUC in plasma and degree of leucopenia was described with a sigmoidal maximum-effect (Emax) model. CONCLUSIONS: The pharmacokinetics of mitomycin during HIPEC could be fitted successfully to a multicompartment model. Relationships between plasma exposure and haematological toxicity were quantified. The developed pharmacokinetic pharmacodynamic model can be used to simulate different dosage schemes in order to optimise mitomycin administration during HIPEC. PMID- 14748624 TI - Prophylactic stenting of the inferior vena cava before transcatheter embolization of renal cell carcinomas: an alternative to filter placement. AB - PURPOSE: To report the use of the self-expanding Wallstent as an alternative to prophylactic inferior vena cava (IVC) filter placement before embolization of renal carcinomas with tumor thrombus. CASE REPORTS: Two patients, a 71-year-old man and an 88-year-old woman, were diagnosed with extensive tumor infiltration of the IVC secondary to renal cell carcinomas. Prophylactic placement of an IVC filter before transcatheter embolization was unsuccessful in both cases; a reduced space for deployment would have left part of the filter inside the right atrium. Instead, a Wallstent was used to constrain the tumor thrombus against the vessel wall and, at the same time, protect the patency of the contralateral kidney. Adequate patencies were confirmed 9 months after stenting in the first patient and after 19 days in the second patient. There were no clinical manifestations of pulmonary embolism. CONCLUSIONS: Wallstent implantation is an alternative prophylactic measure before transarterial embolization of renal carcinomas if IVC filters cannot be placed. PMID- 14748622 TI - Growth, immortalization, and differentiation potential of normal adult human proximal tubule cells. AB - Human proximal tubule epithelial cell lines are potentially useful models to elucidate the complex cellular and molecular details of water and electrolyte homeostasis in the kidney. Samples of normal adult human kidney tissue were obtained from surgical specimens, and S1 segments of proximal convoluted tubules were microdissected, placed on collagen-coated culture plate inserts, and cocultured with lethally irradiated 3T3 fibroblasts. Primary cultures of proximal tubule epithelial cells were infected with a replication-defective retroviral construct encoding either wild-type or temperature-sensitive simian virus 40 large T-antigen. Cells forming electrically resistive monolayers were selected and expanded in culture. Three cell lines (HPCT-03-ts, HPCT-05-wt, and HPCT-06 wt) were characterized for proximal tubule phenotype by electron microscopy, electrophysiology, immunofluorescence, Southern hybridization, and reverse transcriptase-polymerase chain reaction. Each of the three formed polarized, resistive epithelial monolayers with apical microvilli, tight junctional complexes, numerous mitochondria, well-developed Golgi complexes, extensive endoplasmic reticulum, convolutions of the basolateral plasma membrane, and a primary cilium. Each exhibited succinate, phosphate, and Na,K- adenosine triphosphatase (ATPase) transport activity, as well as acidic dipeptide- and adenosine triphosphate-regulated mechanisms of ion transport. Transcripts for Na(+)-bicarbonate cotransporter, Na(+)-H(+) exchanger isoform 3, Na,K-ATPase, parathyroid hormone receptor, epidermal growth factor receptor, and vasopressin V2 receptor were identified. Furthermore, immunoreactive sodium phosphate cotransporter type II, vasopressin receptor V1a, and CLIC-1 (NCC27) were also identified. These well-differentiated, transport-competent cell lines demonstrated the growth, immortalization, and differentiation potential of normal, adult, human proximal tubule cells and consequently have wide applicability in cell biology and renal physiology. PMID- 14748625 TI - Complicated acute aortic dissection type B caused by femoral cannulation for endoscopic coronary artery bypass surgery. AB - PURPOSE: To report endovascular treatment of a patient with complicated acute aortic type B dissection caused by cannulation of the femoral artery for endoscopic coronary artery bypass (CAB) surgery. CASE REPORT: A 63-year-old man underwent single-vessel CAB surgery facilitated by the Da Vinci surgical system. After the procedure, acute lower limb ischemia led to the discovery of a type B dissection from the left subclavian artery to the iliac arteries. The subclavian artery was also dissected, with only minimal flow in the left mammary artery. The visceral and renal vessels originated from the false lumen and were not perfused. Emergent stent-graft placement was undertaken to close the entry tear and re establish perfusion of the lower extremity as well as visceral and renal organs. Additional stent-grafts were placed in the dissected left subclavian artery to perfuse the left internal mammary bypass and in the inferior mesenteric artery, which arose from the false lumen. Six months after operation, the patient is well. CONCLUSIONS: Acute iatrogenic aortic dissection secondary to endoscopic CAB surgery can be treated emergently to restore distal perfusion to vital organs. PMID- 14748626 TI - Comparison of surgical versus endovascular occlusion models in pig femoral arteries. AB - PURPOSE: To compare an endovascular technique with a well established surgical approach to achieve long-term occlusions of large porcine arteries while preserving the integrity of periarterial tissue. METHODS: The femoral arteries in 11 pigs were occluded using surgical techniques on one side and blinded stent grafts in the contralateral vessel. Feasibility, safety, primary and long-term success, and the extent of vascularization were determined over a 3-month period by conventional angiography and histological analysis. A subgroup of animals (n=5) was treated with a locally administered plasmid coding for vascular endothelial growth factor (pVEGF165) to compare both occlusion techniques under conditions of collateral growth induction. RESULTS: The primary and long-term success rates for both occlusion models were 100%. Surgical occlusion of arteries resulted in a significant amount of scar dehiscence and local groin infection compared to the endograft-occluded side. There was no significant difference in capillary densities and collateralization of periarterial areas in a comparison of the occlusion technique: the cross-sectional area of the superficial femoral artery (SFA) was 300 +/- 24 mm2 for endovascular occlusion versus 320 +/- 23 mm2 for surgical occlusion (p=0.559). In the profunda femoris artery, respective values were 418 +/- 35 and 448 +/- 18 mm2 (p=0.474). The local delivery of pVEGF165 resulted in a significant increase in collateral growth in both occlusion models with comparable neovascularization: cross-sectional SFA area increased from 310 +/- 16 to 428 +/- 13 mm2 (p<0.0001); in the PFA, the area increased from 422 +/- 19 to 658 +/- 49 mm2 (p<0.0001). CONCLUSIONS: Endovascular arterial occlusions using blinded stent-grafts allow easy and safe creation of long-term occlusions. Previously described collateralization following surgical occlusions was not observed, indicating that those collaterals may be associated with wound healing rather than ischemia. The occlusion of arteries using blinded stent-grafts in pigs may therefore be an appropriate model for assessing the effects of angiogenic factors in vivo. PMID- 14748627 TI - Impact of symptomatic peripheral arterial disease on 1-year mortality in patients undergoing percutaneous coronary interventions. AB - PURPOSE: To determine the impact of symptomatic peripheral arterial disease (PAD) on clinical outcomes in patients treated with percutaneous coronary interventions (PCI). METHODS AND RESULTS: Symptomatic PAD was identified in 1969 (18.9%) of 10440 consecutive patients undergoing PCI. Patients with PAD were older, more frequently female, and had smaller body surface area and more atherosclerotic risk factors, chronic renal insufficiency, and heart failure. Patients with PAD had lower rates of procedural success (94.2% versus 96.2%, p<0.0001) and higher rates of in-hospital complications, including all-cause mortality (2.1% versus 1.1%, p=0.0002), cardiac death (1.5% versus 0.7%, p=0.0009), urgent coronary artery bypass grafting (1.9% versus 1.2%, p=0.01), recurrent ischemia (5.6% versus 2.8%, p<0.0001), re-PCI to the target lesion (2.4% versus 1.1%, p<0.0001), stroke (0.6% versus 0.3%, p=0.0344), transient ischemic attack (0.4% versus 0.1%, p=0.01), femoral hematoma (10.3% versus 8.5%, p=0.01), retroperitoneal hematoma (0.8% versus 0.3%, p=0.009), limb ischemia (3.0% versus 0.7%, p<0.0001), gastrointestinal bleeding (1.9% versus 0.9%, p<0.0001), and blood transfusion (10.1% versus 5.2%, p<0.0001). At 1-year follow-up, patients with PAD had a higher mortality rate (13.6% versus 5.2%, p<0.0001), a higher rate of myocardial infarction (8.3% versus 6.5%, p=0.008), and also more target lesion (21.2% versus 19.8%, p=0.02) or target vessel revascularization (24.6% versus 21.2%, p=0.002). By multivariate analysis, PAD was an independent predictor of 1-year mortality (odds ratio 1.71, 95% confidence interval 1.42 to 2.07, p<0.0001). CONCLUSIONS: Nearly a fifth of patients undergoing PCI have symptomatic PAD. The presence of PAD is associated with lower rates of procedural success, higher rates of in hospital and 1-year adverse events, and is independently associated with increased 1-year mortality. PMID- 14748628 TI - Carotid stenting for radiation-induced extracranial carotid artery occlusive disease: efficacy and midterm outcomes. AB - PURPOSE: To investigate the immediate and midterm results of carotid stenting for severe radiation-induced extracranial carotid artery disease. METHODS: Between April 1998 and May 2002, 16 patients (15 men; mean age 64 +/- 8 years, range 48 72) presented with 18 severe radiation-induced carotid stenoses in the internal carotid artery (n=3), common carotid artery (n=7), and both vessels (n=8). Thirteen (76%) patients were symptomatic; the mean degree of carotid stenosis was 85% +/- 10% (range 70%-95%). An independent neurological specialist assessed perioperative neurological complications before and after treatment. The patients were followed prospectively for at least 12 months by clinical examination and serial duplex ultrasound scanning. Restenosis was defined as a diameter reduction >50%. RESULTS: Of 18 stent procedures attempted (2 staged), 1 was abandoned owing to failure to pass the guidewire across a tight lesion (94% technical success by intent to treat). In the 17 successfully completed procedures, 17 Wallstents and 4 SMART stents were deployed with satisfactory anatomical results. One postoperative stroke occurred as a result of thromboembolism to the ipsilateral middle cerebral artery and led to hospital death (5.9% combined stroke and death rate). One transient ischemic attack occurred (11.6% neurological event rate). With a median 30-month follow-up (range 5-55), 3 (17.6%) recurrent stenoses (>50%) were detected on duplex scan; 1 repeat angioplasty was performed. No new neurological event has been detected. CONCLUSIONS: Carotid stenting may be performed in patients with irradiation-induced carotid stenosis with acceptable risks and midterm durability. PMID- 14748629 TI - AneuRx stent-graft failure four years after TAA exclusion. AB - PURPOSE: To present a case of graft material disruption 4 years after thoracic aortic aneurysm (TAA) exclusion with the AneuRx stent-graft. CASE REPORT: In 1999, a 62-year-old man underwent successful exclusion of a descending 98-mm TAA with 2 AneuRx stent-grafts. A type I distal endoleak was diagnosed 1 month later, and a distal extension cuff was placed just proximal to the celiac trunk. One year later, a new endoleak was discovered (presumed to be type II); the patient refused proposed thrombin injection to seal the leak. The diameter of the aneurysm enlarged gradually, and in 2003, he presented with a ruptured TAA and a massive left-sided hemathorax. The patient underwent urgent open surgical repair using extracorporeal circulation; the 3 stent-grafts were removed. Unfortunately, the patient died from cardiac failure 2 days later. Macroscopic examination showed several disruptions of the graft fabric and a stent fracture, which presumably explains the endoleak and rupture. CONCLUSIONS: This case reaffirms the necessity of long-term stent-graft surveillance. PMID- 14748630 TI - Endoluminal stent-graft repair of aortobronchial fistulas: bridging or definitive long-term solution? AB - PURPOSE: To describe our experience with endoluminal stent-graft repair of aortobronchial fistulas (ABF) and to analyze midterm results focusing on late chronic graft infections, secondary conversion, and survival. METHODS: The records of 8 patients (6 men; mean age 69 years, range 28-88) treated between March 1997 and October 2003 for traumatic and postsurgical ABFs were reviewed. Seven presented with hemoptysis and 1 with hemorrhagic shock. According to the severity of emergency, patients underwent computed tomography, angiography, bronchoscopy, and transesophageal echocardiography. Preoperatively, no clinical signs of infection were evident. Two different stent-graft models (Talent and Excluder) were implanted using standard endovascular techniques. RESULTS: Procedural and clinical success was achieved in all patients. Paraplegia, secondary intervention, conversion, or procedure-related death was not observed. Mean follow-up was 30 months (range 0.6-77). One patient with a postsurgical ABF (Dacron tube graft) successfully treated with an Excluder stent-graft died 13 months later from hemorrhage secondary to aortoesophageal fistula repair procedures. A second patient died from pneumonia after 42 months. A third patient, in whom 2 Talent stent-grafts had been implanted to treat an ABF from the false lumen of a type B dissection, died 7 months later from massive hemorrhage. CONCLUSIONS: Endoluminal stent-grafting of ABF is feasible and the preferred method of treatment. Secondary conversion due to endograft infection is not absolutely mandatory, but close surveillance is necessary. PMID- 14748631 TI - How many patients with infrarenal aneurysms are candidates for endovascular repair? The Northern California experience. AB - PURPOSE: To determine how many patients with abdominal aortic aneurysms (AAA) meet the anatomical selection criteria for AneuRx stent-graft repair in community hospitals of Northern California. METHODS: The records were reviewed of 220 AAA patients (171 men, 49 women) who were considered for endovascular repair by the treating vascular surgeon at 28 community hospitals in Northern California between January and October 2001. Contrast computed tomographic angiography (CTA) and selective arteriography were performed at each institution and reviewed by a centralized, independent image-reading center. Selection criteria determined by the manufacturer and published in the indications for use were applied to each set of imaging studies. The number of patients who met inclusion criteria were recorded, as were the anatomical characteristics of each aneurysm. RESULTS: The mean aneurysm size in the 220 patients was 55.3 +/- 0.7 mm. Among these patients, 122 (55%) were judged to be candidates for endovascular repair and 98 (45%) were considered ineligible. The primary anatomical reason for ineligibility was a short infrarenal neck in 43 (44%) patients, followed by a large proximal neck diameter (25, 25%), iliac aneurysms (10, 10%), extremely tortuous or calcified neck (7, 7%), iliac occlusion (6, 6%), and small distal aortic bifurcation and accessory renal arteries (5, 5%). Four (4%) patients were classified as non candidates due to poor quality imaging. There was no difference in aneurysm diameter (54.0 +/- 0.8 versus 57.1 +/- 1.2 mm, p=NS) or age (72.2 +/- 1.2 versus 74.6 +/- 2.2 years, p=NS) between candidates and non-candidates. However, proportionally more men (60%) than women (39%) were eligible for endovascular repair with the AneuRx stent-graft (p<0.05). All 122 patients who were considered candidates for endovascular repair were treated, with successful stent-graft placement achieved in 121 (99%). CONCLUSIONS: Fifty-five percent of patients considered for endovascular AAA repair in community hospitals in Northern California met the anatomical selection criteria for the AneuRx stent-graft. Men appeared to be twice as likely to meet the eligibility requirements as women. Unfavorable infrarenal neck anatomy was the primary exclusion criterion for endovascular repair in this community setting. PMID- 14748632 TI - Stent-graft placement in atherosclerotic descending thoracic aortic aneurysms: midterm results. AB - PURPOSE: To determine midterm durability and need for reinterventions after stent graft placement in atherosclerotic descending thoracic aortic aneurysms. METHODS: Fifty-four patients (38 men; mean age 68 years, range 33-87) underwent stent graft repair of chronic atherosclerotic aneurysms of the descending thoracic aorta between November 1996 and December 2002. Acute aortic syndromes (type B aortic dissections, perforating ulcers, and traumatic dissections) were excluded from analysis. Two types of commercially available stent-grafts were used (Talent and Excluder). RESULTS: The primary technical success rate was 94.4%. In-hospital mortality was 3.7% (2/54). No adverse neurological events were encountered. Of 3 (5.6%) early type I endoleaks, 2 (3.7%) required reintervention; the other type I endoleak closed spontaneously. Mean follow-up was 38 months (range 1-72) in the 52 surviving patients. Four (7.7%) type I, 7 (13.5%) type II, and 4 (7.7%) type III endoleaks were seen. Three 3 patients had additional treatment for endoleaks (proximal stenting [type Ia], open thoracoabdominal repair [type Ib], and embolization [type II]). Two of the 3 reinterventions were performed in the first year of follow-up. One additional patient with a type Ia endoleak not suitable for reintervention is under close observation. There were no differences in the number of endoleaks between the 2 types of stent-grafts. Three-year event-free survival was 63%. CONCLUSIONS: Midterm durability of endovascular stent-graft placement in atherosclerotic descending aortic aneurysms seems promising, as the rate of endoleaks requiring reintervention is acceptably low. In our series, endoleak formation occurred during the first year after stent-graft placement, so close follow-up of patients after aortic aneurysm repair is crucial. Further studies are warranted to evaluate long-term durability of this new treatment modality. PMID- 14748633 TI - Diagnostic value of CT virtual intravascular endoscopy in aortic stent-grafting. AB - PURPOSE: To investigate the diagnostic value of postprocessing techniques for 3 dimensional (3D) computed tomography (CT), with emphasis on CT virtual intravascular endoscopy (VIE), in patients with abdominal aortic aneurysms (AAA) treated with suprarenal stent-grafts. METHODS: The preprocedural and postprocedural CT datasets from 47 AAA patients (40 men; mean age 75 years, range 61-87) undergoing aortic stent-grafting with suprarenal fixation were examined. The CT datasets were processed to create various 3D reconstructions: shaded surface display (SSD), maximum intensity projection (MIP), and VIE. Three independent radiologists assessed various diagnostic parameters for each 3D reconstruction method and compared them to axial CT images. RESULTS: Scores for VIE reconstructions were inferior to axial CT images in the visualization of normal arterial branches, measurement of the aneurysm diameter and neck length, as well as assessment of vessel patency and presence of endoleaks. VIE was rated superior to axial CT and other 3D imaging methods in visualizing the configuration of stent struts relative to the aortic branch ostia and the number of stent wires crossing the ostia in >80% of cases. CONCLUSIONS: VIE was not found to play a role in most preoperative situations compared to axial CT images. However, VIE provided additional postgrafting information on the 3D relationship of the suprarenal stent struts to the aortic branch ostia (in particular the renal and superior mesenteric arteries). VIE findings might aid clinicians in accurately assessing the effect of suprarenal stent-grafting on the renal arteries. PMID- 14748634 TI - Serial CT volume and thrombus length measurements after endovascular repair of Stanford type B aortic dissection. AB - PURPOSE: To evaluate the outcome of stent-graft placement in Stanford type B aortic dissection using contrast-enhanced spiral computed tomographic (CT) measurements of true and false lumen volumes and thrombus length. METHODS: Among 18 consecutive patients (13 men; mean age 60 years, range 44-79) who underwent endovascular repair of Stanford type B dissection, 12 completed at least a 12 month follow-up, which included CT measurements of true and false lumen volumes and thrombus lengths prior to discharge and at 6 and 12 months postimplantation. Volumes were assessed in 3 different aortic segments (A1, A2, A3) extending from the proximal attachment site of the prosthesis to the aortic bifurcation. In addition, thrombus length was measured to evaluate the influence of clot formation on outcome of the false lumen volume. RESULTS: Mean follow-up was 27 months (range 12-60). Within 12 months, mean true lumen volumes showed statistically significant increases in the A1 (p<0.001) and A2 (p=0.003) segments; false lumen volumes showed a significant decrease in the A1 segment (p=0.002) but an insignificant increase in the A2 segment. No substantial volume changes were observed in the A3 segment. Extension of clot formation in the false lumen varied among patients and over time. Length of stent-grafts, percentage of stented dissection length, or visceral arteries originating from the false lumen did not significantly influence thrombus development, nor did these parameters or thrombus formation distal to the prosthesis have a relationship to false lumen volumes. CONCLUSIONS: Volumetric analysis after endovascular repair of Stanford type B dissection shows optimal technical outcome in the stented segment, whereas the false lumen in the segment immediately adjacent to the stent-graft seems to be a vulnerable area. Extension of clot formation beyond the endograft seems to be no reliable predictor of outcome. PMID- 14748635 TI - The dynamics of interlip coupling in speakers with a repaired unilateral cleft lip history. AB - In this paper, data are presented on individual movement characteristics of the upper and lower lip and on interlip coordination in speakers with a repaired unilateral cleft upper lip history and age-matched control participants. The data were acquired using the AG100 EMMA system while the participants produced a selection of nonspeech and speech tasks. The participants with a repaired unilateral cleft upper lip history showed reduced upper-lip movement ranges and peak velocities and a more variable spatiotemporal pattern for individual upper lip movement cycles, in addition to a more variable interlip coupling. The latter difference also proved to be more pronounced for the younger speakers with a repaired cleft upper lip. Overall, for both groups, the linguistically more complex task showed more variability in the individual upper- and lower-lip movement cycles and their coupling. In the discussion, we address the potential relationships between the kinematic data for upper lip in the repaired-cleft-lip speakers and the findings on movement and coordination stability as they might fit within current notions of coordination dynamics theory. PMID- 14748636 TI - Characteristics of speech disfluency and stuttering behaviors in 3- and 4-year old children. AB - The purpose of this investigation was to quantitatively and qualitatively characterize speech disfluencies exhibited by 3- and 4-year-old children who do (CWS, N = 36) and do not (CWNS, N = 36) stutter. Five measures of speech disfluency (e.g., percentage of total, other, and stuttering-like disfluencies, mean number of repetition units, and weighted SLD measure) were used in attempts to differentiate CWS from CWNS. Similar measures of stuttering (e.g., percentage of stuttering-like disfluencies consisting of disrhythmic phonations) were used to characterize speech disfluencies in 3- and 4-year-old CWS in relation to time since stuttering onset (TSO). It was hypothesized that such measures of speech disfluency should significantly differ between CWS and CWNS, as well as 3- versus 4-year-old CWS in relation to TSO. Results indicated that 4 out of the 5 dependent measures significantly differed between CWS and CWNS, and within the CWS group there was a significant relationship between TSO and the percentage of stuttering-like disfluencies when the effects of chronological age were partialled out of the regression analyses. Furthermore, 4-year-old CWS exhibited a moderate correlation between TSO and the percentage of stuttering-like disfluencies consisting of disrhythmic phonations, whereas 3-year-old CWS exhibited no such relationship between these two variables. Findings were taken to suggest that certain measures of speech disfluency appreciably differentiate CWS from CWNS and that 4-year-old CWS exhibit changes in nonreiterative forms of stuttering as a function of time since stuttering onset. PMID- 14748637 TI - Acoustic and perceptual consequences of articulatory rate change in Parkinson disease. AB - This study sought to characterize the relationship among voluntary rate modification, vocal tract acoustic output, and perceptual impressions of speech for individuals with idiopathic Parkinson disease (PD). Healthy control speakers were studied for comparison purposes. Four research questions were addressed: (1) How is rate modification evidenced in acoustic measures of segmental and global timing? (2) What is the impact of rate modification on measures of acoustic working space for select vowels and consonants? (3) What is the impact of rate modification on perceptual impressions of severity? (4) Are rate-induced changes in measures of acoustic working space related to perceptual impressions of severity? Speakers read the Farm Passage at habitual, slow, and fast rates. The vowels /i/, /ae/, /u/, and /A/ and the fricatives /s/ and /S/ were of interest. Acoustic measures included articulatory rate, segment durations, vowel formant frequencies, and first moment coefficients. Measures of acoustic working space for vowels and fricatives also were derived. The results indicated that temporal acoustic measures changed in the expected direction across rate conditions, with a tendency toward slightly faster rates for the PD group. In addition, the relative rate change for the Fast and Slow conditions compared to the Habitual condition was similar across groups. Rate did not strongly affect measures of acoustic working space for the PD group as a whole, but there was a tendency for slower rates to be associated with larger measures of acoustic working space. Finally, there was not a strong relationship between perceived severity and measures of acoustic working space across the rate continuum for either group. Rather, the relationship between perceived severity and measures of acoustic working space was such that the PD group exhibited smaller measures of acoustic working space and more severe perceptual estimates than the control speakers, irrespective of rate condition. PMID- 14748638 TI - Muscle, adipose, and connective tissue variations in intrinsic musculature of the adult human tongue. AB - The purpose of this investigation was to identify the composition and organization of lingual tissues underlying the histostructural and biomechanical functions of the adult human tongue. The small-scale structures of three intrinsic muscle regions, their principal cells, structural complexities, and differences in underlying tissue composition were compared to other skeletal muscle systems and the results discussed in relation to lingual morphology. Analysis of pixel color distributions determined the percent area concentration of each stained tissue component. Results indicated that muscle content increased from anterior to posterior (p <.0001). Greater adipose (p =.005) and connective tissue (p <.002) concentrations occurred in anterior regions. Dense collagen sheaths and elastic fibers found anteriorly occurred with less magnitude in medial and posterior sites. The unique elastin, collagen, and adipose connective tissue distributions found in intrinsic sampling sites are discussed in terms of understanding lingual biomechanics in both normal and pathologic states. PMID- 14748640 TI - Direct magnitude estimation and interval scaling of hypernasality. AB - Hypernasality is most commonly assessed using equal-appearing interval (EAI) scaling. Recently, the validity of EAI scaling for the evaluation of hypernasality has been questioned. The issue of validity rests on the psychophysical nature of the dimension to be rated. The purpose of this study was to compare EAI scaling with direct magnitude estimation (DME), in order to determine whether EAI scaling is a valid procedure for the evaluation of hypernasality. Connected speech samples from 20 individuals with repaired cleft palate and hypernasality were used. Twenty listeners undertook the listening tasks, which included EAI scaling, DME with modulus (DME-M), and DME without modulus (DME-WM). The results showed a curvilinear relationship between EAI and DME-M and between EAI and DME-WM, suggesting that EAI may not be a valid method for the evaluation of hypernasality; DME is recommended. PMID- 14748639 TI - The sequential development of jaw and lip control for speech. AB - Vertical displacements of the upper lip, lower lip, and jaw during speech were recorded for groups of 1-, 2-, and 6-year-olds and adults to examine if control over these articulators develops sequentially. All movement traces were amplitudeand time-normalized. The developmental course of upper lip, lower lip, and jaw control was examined by quantifying age-related changes in the similarity of each articulator's movement patterns to those produced by adult subjects and by same-age peers. In addition, differences in token-to-token stability of articulatory movement were assessed among the different age groups. The experimental findings revealed that 1- and 2-year-old children's jaw movements were significantly more adult-like than their upper and lower lip movements, which were more variable. In contrast, upper and lower lip movement patterns became more adult-like with maturation. These findings suggest that the earliest stages of speech motor development are constrained by the nonuniform development of articulatory control, with the jaw preceding the lips. The observed developmental patterns suggest that the properties of the oral motor control system significantly influence the pattern of speech sound acquisition. PMID- 14748641 TI - Speaking and breathing in high respiratory drive. AB - Pulmonary ventilation during speech breathing reflects the sum of the airflow changes used to speak and to meet the metabolic needs of the body. Studying interactions between speaking and breathing may provide insights into the mechanisms of shared respiratory control. The purposes of this study were to determine if healthy subjects exhibit task-specific breathing behaviors in high respiratory drive and to document subjects' perceptions during breathing and speaking under these conditions. Ten men were studied in air and high CO2. Magnetometers were used to estimate lung volume, rib cage and abdomen volumes, minute volume, breathing frequency, tidal volume, inspiratory and expiratory duration, and inspiratory and expiratory flow. Subjects' perceptions were assessed informally. Results indicated that the chest wall kinematic behaviors associated with breathing and speaking in high drive were similar in pattern but differed in the magnitudes of lung volume and rib cage volume events and in inspiratory and expiratory flow. Linguistic influences remained strong, but not as strong as under normal conditions. All subjects reported a heightened sense of breathing-related discomfort during speaking as opposed to breathing in high respiratory drive. We conclude that in healthy subjects breathing behavior associated with speaking in high respiratory drive is guided continuously by shared linguistic and metabolic influences. A parallel-processing model is proposed to explain the behaviors observed. PMID- 14748642 TI - Longitudinal changes in articulation rate and phonetic phrase length in children with speech delay. AB - This study examined long-term changes in articulation rate (the pace at which speech segments are produced) and phonetic phrase length in the conversational speech of two groups of children with speech delay (SD) of unknown origin. Initial testing for both groups occurred at preschool age, with follow-up testing conducted for the Early Follow-Up Group (n = 17) at age 9 years and for the Late Follow-Up Group (n = 36) at age 12-16 years. At follow-up testing both groups produced significantly faster articulation rates (measured in both syllables per second and phones per second) and significantly longer phonetic phrases (measured in both syllables and phones) than at initial testing. Articulation rates at both test times were also judged to be similar to published values from typically developing children of similar ages when measured in syllables per second. However, findings for rate in phones per second suggested that at least at initial testing the children were articulating speech at a slower rate than their typically developing peers. This latter finding, however, may have been an artifact of the high frequency of errors--such as cluster reduction and final consonant deletion--observed in the initial samples. It would appear, therefore, that children with SD of unknown origin may start out with slower than normal articulation rates but eventually catch up to their typically developing peers. PMID- 14748643 TI - The effect of anchors and training on the reliability of perceptual voice evaluation. AB - Perceptual voice evaluation is a common clinical tool for rating the severity of vocal quality impairment. However, the evaluation process involves subjective judgment, and reliability is therefore a major issue that needs to be considered. When listeners are asked to judge the quality of a voice signal, they use their own internal standards as the references. These internal standards can be variable, as different individuals may have acquired different standards in prior situations. In order to improve the reliability of the perceptual voice evaluation process, external anchors and training are provided to counteract the effect of these internal standards. This study investigated to what extent the provision of anchors and a training program would improve the reliability of perceptual voice evaluation by naive listeners. The results show, in general, that anchors and training helped to improve the reliability of perceptual voice evaluation, especially in the rating of male voices. Furthermore, it was found that anchors made up of synthesized signals combined with training were more effective in improving reliability in judging perceptual roughness and breathiness than natural voice anchors. PMID- 14748644 TI - Defining and measuring speech movement events. AB - A long-held view in speech research is that utterances are built up from a series of discrete units joined together. However, it is difficult to reconcile this view with the observation that speech movement waveforms are smooth and continuous. Developing methods for reliable identification of speech movement units is necessary for describing speech motor behavior and for addressing theoretically relevant questions about its organization. We describe a simple method of parsing movement signals into a series of individual movement "strokes," where a stroke is defined as the period between two successive local minima in the speed history of an articulator point, and use that method to segment speech-related movement of marker points placed on the tongue blade, tongue dorsum, lower lip, and jaw in a group of healthy young speakers. Articulator fleshpoints could be distinguished on the basis of kinematic features (i.e., peak and boundary speed, duration and distance) of the strokes they produce. Further, tongue blade and jaw fleshpoint strokes identified to temporally overlap with acoustic events identified as alveolar fricatives could be distinguished from speech strokes in general on the basis of a number of kinematic measures. Finally, the acoustic timing of alveolar fricatives did not appear to be related to the kinematic features of strokes presumed to be related to their production in any direct way. The advantages and disadvantages of this simple approach to defining movement units are discussed. PMID- 14748645 TI - Lexical organization and phonological change in treatment. AB - Word frequency and neighborhood density are properties of lexical organization that differentially influence spoken-word recognition. This study examined whether these same properties also affect spoken-word production, particularly as related to children with functional phonological delays. The hypothesis was that differential generalization would be associated with a word's frequency and its neighborhood density when manipulated as input in phonological treatment. Using a multiple baseline across subjects design, 8 children (aged 3;10 to 5;4) were randomly enrolled in 1 of 4 experimental conditions targeting errored sounds in high-frequency, low-frequency, high-density, or low-density words. Dependent measures were generalization of treated sounds and untreated sounds within and across manner classes as measured during and following treatment. Results supported a hierarchy of phonological generalization by experimental condition. The clinical implications lie in planning for generalization through the input presented in treatment. Theoretically, the results demonstrate that lexical organization of words in the mental lexicon interacts with phonological structure in learning. PMID- 14748646 TI - An evaluation of the facilitative effects of inverted yes-no questions on the acquisition of auxiliary verbs. AB - According to the Auxiliary Clarification Hypothesis (ACH), yes-no questions with sentence-initial auxiliaries (i.e., inverted questions) facilitate children's initial acquisition of auxiliary verbs. Sixteen 3-year-old children with specific language impairment (SLI) and 18 2-year-olds with typical language (TL) participated in an experiment to evaluate the ACH. The children were not yet making use of auxiliaries. Half of the children participated in twenty 30-min "enrichment" sessions over a 2-month period, during which an assistant produced 30 inverted question recasts in response to the child's own utterances. Fifteen question recasts contained the auxiliary is, and 15 contained the modal will. The other half of the children participated in play sessions but were not exposed to inverted is and will questions. Contrary to predictions based on the ACH, the results revealed no positive effects of the enrichment for is, for will, or for the broader BE and Modal auxiliary categories for either group of children. The children with TL demonstrated advantages over the children with SLI for the general category of BE forms but not for the category of Modals. Inverted questions may be too complex to foster the initial acquisition of auxiliaries in children not already using them productively. PMID- 14748647 TI - Narrative content as described by individuals with Down syndrome and typically developing children. AB - Narratives of the wordless picture story, Frog, Where Are You?, by 33 individuals with Down syndrome and typically developing children (33 matched for mental age, 33 for syntax comprehension, 33 for mean length of utterance) were analyzed for expression of plot line, story theme, and the protagonists' misadventures in the story. Despite their restricted expressive syntax and vocabulary, the group with Down syndrome expressed more plot line and thematic content and more of one of the protagonists' misadventures than the MLU controls; they most resembled the syntax comprehension control participants. We conclude that the group with Down syndrome had a conceptual understanding of the picture story similar to that of the TACL-R group and a strategy for expressing that understanding despite expressive lexical and syntactic limitations; this resulted in the expression of more narrative content than formal measures of expressive language would predict. We propose that the higher syntactic comprehension skills of the group with Down syndrome, combined with their experience with story content (listening to stories), may have contributed to their developing higher-level story schemas than would be expected given their MLUs. PMID- 14748649 TI - Understanding bilingualism and its impact on speech audiometry. AB - This tutorial provides a review of auditory research conducted with monolingual and bilingual speakers of Spanish and English. Based on a functional view of bilingualism and on auditory research findings showing that the bilingual experience may affect the outcome of auditory research, we discuss methods for improving descriptions of linguistically diverse research participants. The review delves into how the bilingual experience can affect auditory research outcomes and discusses ways in which experimental design can be adjusted when bilingual or monolingual participants are used for research needs. The goal of the tutorial is to increase awareness about the complexities of using bilinguals in auditory research, thereby improving the quality of auditory research involving bilingual research participants. PMID- 14748648 TI - Stability of behavioral ratings of children with SLI. AB - Standardized rating scales represent the most reliable method of identifying socioemotional behavioral problems in children with SLI. However, limited information exists on the situational specificity or stability of rating scales applied to this population. In Redmond and Rice (1998), we presented evidence of limited reliability and stability between ratings collected from teachers and parents during kindergarten and first grade. In this research report, we provide additional data on the same group of children over the early elementary period (kindergarten-second grade). The results indicate diminishment in teacher reported behavior problems in most areas of socioemotional development from kindergarten to second grade and increasing congruence between teacher and parent ratings. PMID- 14748650 TI - Children's recognition of cartoon voices. AB - We examined developmental changes in talker recognition skills by assessing 3-, 4 , and 5-year-old children's recognition of 20 cartoon characters' voices. For each participant, the character set was subdivided into more and less familiar talkers based on the participant's ability to name each character. Four- and 5- year-old children recognized more of the voices (81% and 86%, respectively) than did 3-year-olds (61%), although performance of all age groups was well above chance. All groups of children were more accurate at recognizing more familiar than less familiar characters. These results suggest that indexical information about a talker becomes an integral part of the perceptual record in memory and can be used by children at a very young age. These results are important because children's ability to learn vocal sources may be an important aid to the development of spoken word recognition. PMID- 14748652 TI - Gefitinib (Iressa): a novel treatment for non-small cell lung cancer. AB - Current standard therapy for advanced or metastatic non-small cell lung cancer (NSCLC) includes chemotherapy, which is often associated with limited efficacy and toxicity. Thus, there is an unmet need for novel anticancer agents that are both effective and well tolerated in patients with NSCLC. Gefitinib (Iressa) is an orally active epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor in advanced clinical development, and the first agent of its kind to receive approval. Extensive evidence indicates that gefitinib provides significant antitumor activity in previously treated advanced NSCLC, along with fast symptom improvement. This orally administered agent is generally well tolerated. Gefitinib has also shown promising efficacy in other solid tumors that rely on EGFR-related mechanisms for growth and survival. This article reviews the profile of gefitinib and the evidence supporting its use in the treatment of NSCLC. PMID- 14748653 TI - Iodine-131 tositumomab (Bexxar): radioimmunoconjugate therapy for indolent and transformed B-cell non-Hodgkin's lymphoma. AB - Tositumomab is an immunoglobulin G murine monoclonal antibody that binds to the CD20 antigen on the surface of normal and malignant human B-cells. Tositumomab is linked covalently with iodine-131 to produce the radioimmunoconjugate iodine-131 tositumomab (Bexxar). The iodine-131 tositumomab regimen was approved by the US Food and Drug Administration in June 2003 for the treatment of patients with CD20 positive, follicular non-Hodgkin's lymphoma, both with and without transformation, whose disease is refractory to rituximab (Rituxan) and has relapsed following chemotherapy. The dose-limiting toxicity of iodine-131 tositumomab is bone marrow suppression and resulting cytopenias. Unlike chemotherapy, the majority of nonhematologic adverse events associated with iodine-131 tositumomab are mild to moderate in nature and usually self limited. Iodine-131 tositumomab represents one of the most active single agents for the treatment of recurrent indolent and transformed B-cell non-Hodgkin's lymphoma, as demonstrated by several clinical trials summarized in this review. At the present time, the use of radioimmunoconjugate therapy is largely limited to patients with disease refractory to rituximab therapy and transformed disease not amenable to high-dose therapy and autologous stem cell support. Longer follow-up of ongoing clinical trials should provide reassurance as to safety and insights as to the additive stem cell toxicity from iodine-131 tositumomab administration. Studies are also addressing the role of iodine-131 tositumomab as a component of initial therapy for indolent non-Hodgkin's lymphoma and in additional histologies of non Hodgkin's lymphoma. PMID- 14748654 TI - Pentostatin (Nipent) in the treatment of chronic lymphocyte leukemia and hairy cell leukemia. AB - Pentostatin (Nipent), formerly known as deoxycoformycin, is a profound inhibitor of the enzyme adenosine deaminase, resulting in the accumulation of metabolites that inhibit ribonucleotide reductase, which in turn inhibits DNA synthesis. Pentostatin was the first of the purine analogs to undergo extensive testing as an anticancer agent and the first to receive US Food and Drug Administration approval for a treatment indication. It is highly effective as first-line monotherapy in hairy cell leukemia, with a complete response rate of 80% and a 10 year survival rate of around 80%. Pentostatin is also active in chronic lymphocyte leukemia as a single agent, but appears even more promising in combination approaches with the alkylating agents chlorambucil or cyclophosphamide. Due to the increasing recognition of delayed severe stem cell and immune suppression following therapy with other purine analogs, there has been renewed interest in pentostatin, especially in combination with chemotherapy and/or the monoclonal antibody rituximab (Rituxan) in chronic lymphocyte leukemia. PMID- 14748655 TI - Bicalutamide (Casodex) in the treatment of prostate cancer. AB - Prostate cancer is an important healthcare issue in men worldwide. With the advent of prostate-specific antigen screening and improved diagnostic techniques, prostate cancer is now being diagnosed in younger men and at earlier disease stages. As a result, patients often live with their disease for many years after diagnosis. This shift in the patient profile has focused attention to the impact of treatment on quality of life. Medical/surgical castration has traditionally been the mainstay of hormonal therapy but is associated with side effects including loss of libido and impotence. Nonsteroidal antiandrogens such as bicalutamide (Casodex) offer an effective alternative to castration with potential quality-of-life benefits. This paper reviews the evidence concerning the use of bicalutamide at all stages of the disease. PMID- 14748656 TI - 11th annual congress of the European Society of Gene Therapy. PMID- 14748657 TI - New anticancer agents and therapeutic strategies in development for solid cancers: a clinical perspective. AB - In addition to well-known chemotherapeutic agents used in the treatment of solid cancers, promising novel cytotoxic agents are being investigated. Among them are analogs of existing cytotoxic agents, aimed at improving the therapeutic index, and new families such as the epothilone compounds. Agents that target the tyrosine kinase-dependent pathways, farnesyl transferase modulators, Raf kinase inhibitors, antisense molecules to Bcl-2 and proteasome modulators, agents that bind to key proteins involved in critical phases of the cell cycle, as well as antiangiogenesis strategies, are all promising approaches in the treatment of solid cancers. The combination of cytotoxics, hormonal agents or radiotherapy with new molecular-targeted therapies represents one of the main strategies to improve survival in solid cancers. A clinical perspective of these agents as monotherapy or combination therapy will be presented in this paper. PMID- 14748658 TI - Targeted therapies in oncology: in the crosshairs or at the crossroads? AB - Normal cellular behavior depends on functional integration of extracellular stimuli with intracellular signal transduction pathways. Coupling cell surface message reception to nuclear gene expression is no longer a linear model constructed with molecular components acting merely as conduits to relay signals that cascade toward the nucleus. What has emerged instead is a highly integrated circuit comprised of numerous molecular components harmoniously programmed to communicate a multitude of internal signals that controls cellular response. Despite increasing understanding of cell signaling, mutinous elements embedded in these pathways have defied complete resolution. Research indicates that propagation of signals emanating from the extracellular environment to the cell nucleus follows a complex internal circuit equipped with sophisticated molecular components that provide rigid control over a variety of cellular responses. Although increasing understanding of genetic aberrations and signaling pathway transgressions can lead to novel strategies for targeting cancer cells, the disappointing results from clinical trials suggest that the occult processes responsible for neoplastic transformation remain largely unexplained. PMID- 14748659 TI - Radiofrequency ablation of small renal tumors. AB - Technological improvements have led to increased diagnosis of small renal tumors and to the development of minimally invasive treatments for these tumors. As a minimally invasive ablative treatment, radiofrequency ablation shows promise as a therapeutic option for selected patients. By creating heat using radiofrequency waves, radiofrequency ablation leads to controlled thermal ablation within the target tissue. Experimental studies have shown radiofrequency ablation to be safe and reproducible. Clinical studies are ongoing but early evidence suggests that radiofrequency ablation is safe and oncologic results are encouraging. With further development of monitoring techniques and additional maturation of oncologic data, radiofrequency ablation may become a standard treatment option for patients with small renal tumors. PMID- 14748660 TI - Clinical and epidemiologic burden of chronic myelogenous leukemia. AB - Chronic myelogenous leukemia represents 7-20% of all leukemia cases, with a worldwide incidence projected at less than one to two per 100,000 people. Approximately 85% of patients are diagnosed with chronic-phase chronic myelogenous leukemia and up to 40% are asymptomatic. Treatment strategies include chemotherapy, interferon-alpha therapy, transplantation (bone marrow/stem cell transplant) and imatinib mesylate (Gleevec), with the impact of treatment best realized during the chronic phase of the disease. Only transplantation has been clinically demonstrated to eradicate the Philadelphia chromosome, alter the natural course of the disease and cure patients diagnosed with chronic myelogenous leukemia. Interferon-alpha is currently considered for first-line treatment, however, the recent introduction of targeted therapy may change clinical practice. Ongoing research focused on new drug combinations and target therapies may eventually expand the armamentarium available to cure this disease. PMID- 14748661 TI - Helicobacter pylori and molecular mechanisms of gastric carcinogenesis: targets for prevention and therapy. AB - Gastric cancer is still a leading cause of cancer death worldwide. It is frequently diagnosed in an advanced stage and the overall prognosis is dismal. A major improvement will only be reached with a better understanding of the genetic changes underlying gastric cancer, which may provide us with effective prevention and treatment strategies. This review will highlight the present knowledge on the role of Helicobacter pylori in gastric carcinogenesis and the molecular pathogenesis of gastric cancer, as well as provide a brief overview of possible future targets for more specific and successful treatment options. PMID- 14748662 TI - Molecular neuro-oncology and development of targeted therapeutic strategies for brain tumors. Part 2: PI3K/Akt/PTEN, mTOR, SHH/PTCH and angiogenesis. AB - Brain tumors are a diverse group of malignancies that remain refractory to conventional treatment approaches. Molecular neuro-oncology has now begun to clarify the transformed phenotype of brain tumors and identify oncogenic pathways that might be amenable to targeted therapy. Activity of the phosphoinositide 3; kinase (PI3K)/Akt pathway is often upregulated in brain tumors due to excessive stimulation by growth factor receptors and Ras. Loss of function of the tumor suppressor gene PTEN also frequently contributes to upregulation of PI3K/Akt. Several compounds, such as wortmannin and LY-294002, can target PI3K and inhibit activity of this pathway. The mammalian target of rapamycin (mTOR) is an important regulator of cell growth and metabolism and is often upregulated by Akt. Clinical trials of CCI-779, an inhibitor of mTOR, are ongoing in recurrent malignant glioma patients. The sonic hedgehog/PTCH pathway is involved in the tumorigenesis of some familial and sporadic medulloblastomas. This pathway can be targeted by cyclopamine, which is under evaluation in preclinical studies. Angiogenesis is a critical process for development and progression of brain tumors. Targeted approaches to inhibit angiogenesis include monoclonal antibodies, receptor tyrosine kinase inhibitors, antisense oligonucleotides and gene therapy. Clinical trials are ongoing for numerous angiogenesis inhibitors, including thalidomide, CC-5103 and PTK 787/ZK 222584. Further development of targeted therapies and evaluation of these new agents in clinical trials will be needed to improve survival and quality of life of patients with brain tumors. PMID- 14748663 TI - Challenges facing antiangiogenic therapy for cancer: impact of the tumor extracellular environment. AB - It is well known that angiogenesis plays an important role in malignant tumor progression. Thus, a great deal of effort has been focused on the development and evaluation of novel angiogenesis inhibitors for the treatment of human malignancies. In this review, the role of angiogenesis in tumor growth will be examined, as well as efforts to develop and use antiangiogenic therapies to treat malignant tumors. In particular, focus will be on the extracellular environment and the challenges of using antiangiogenic therapy in the clinical setting, in terms of toxicities, potential mechanisms of tumor resistance and optimization of clinical trial design. Attention will be focused upon a mechanistic understanding of the variability and dynamic nature of individual tumor microenvironments, and the potential impact this has on antiangiogenic therapies. PMID- 14748664 TI - Multimodality therapy for cancers of the esophagus and gastric cardia. AB - The role of multimodal treatment in the management of esophageal cancer is controversial. There are conflicting results from studies on the effect of neoadjuvant and/or adjuvant treatment on long-term survival. Following a search of the Medline database, the authors examine the results of randomized studies on the various treatment protocols available and discuss future therapeutic improvements. PMID- 14748665 TI - Targeted cancer therapy with gonadotropin-releasing hormone chimeric proteins. AB - Tumor-associated antigens (TAAs) have been identified mainly to determine cancer prognosis. In the past few years, TAAs have been used in the development of treatment modalities such as tumor vaccination. This review describes an additional application of TAAs: as a target for specific antitumor treatment. Since TAAs are overexpressed on the tumor cell surface, they can be targeted to deliver drugs directly to cancer cells. One such delivery system exploits chimeric proteins. Chimeric proteins are a class of targeted molecules designed to recognize and specifically destroy cells that overexpress specific receptors. These molecules, designed and constructed by gene fusion techniques, comprise both cell-targeting and cell-killing moieties. The authors' laboratory has developed a number of chimeric proteins using gonadotropin-releasing hormone (GnRH) as the targeting moiety. These chimeras recognize a GnRH binding site that is expressed on adenocarcinoma cells. GnRH was fused to a large number of killing moieties, including bacterial and human proapoptotic proteins. All GnRH-based chimeric proteins selectively killed adenocarcinoma cells both in vitro and in vivo. Utilizing chimeric proteins for targeted therapy represents a new and exciting therapeutic modality for the treatment of cancer in humans. PMID- 14748666 TI - The "Cam affair": an isolated incident or destined to be repeated? PMID- 14748667 TI - Training our future rural medical workforce. PMID- 14748668 TI - E-drug deals: part of the Wild West world of e-commerce. PMID- 14748669 TI - The continuing legacy of the United Kingdom Prospective Diabetes Study. PMID- 14748670 TI - A case for more year-long internships outside metropolitan areas? AB - OBJECTIVE: To determine whether medical graduates who spent their intern year at a non-metropolitan hospital were more likely to practise outside metropolitan areas on completion of training than were interns in metropolitan hospitals. DESIGN: Retrospective follow-up of doctors who held year-long internships at a non-metropolitan hospital and interns from metropolitan hospitals. SETTING: Ballarat Base Hospital (BBH) (Rural, Remote and Metropolitan Area [RRMA] rural zone) and hospitals in Melbourne and Geelong (RRMA metropolitan zone). PARTICIPANTS: 57/63 (90%) Victorian medical graduates completing internships at BBH between 1989 and 1997 and 126/126 (100%) sex-matched metropolitan interns, chosen at random. MAIN OUTCOME MEASURES: Practice location in 2002. RESULTS: More BBH interns were practising as GPs outside metropolitan areas (44%) than metropolitan interns (13%) (difference, 31%; 95% CI, 17%-45%). The proportion of interns in specialist practice outside metropolitan areas was small for both groups - zero and 3%, respectively (difference, - 3%; 95% CI, - 6% to 0). None of the specialist training posts held by interns were outside metropolitan areas. Of BBH interns entering general practice, 41% (95% CI, 24%-58%) did so in the local health region. CONCLUSIONS: Regional interns are a good source of non metropolitan GPs, especially locally. Prospective studies to determine the precise influence of regional internships on eventual practice location, and whether more such posts would lead to more graduates entering non-metropolitan practice, would be worthwhile. PMID- 14748671 TI - Causes of sudden cardiac death in young Australians. AB - OBJECTIVES: To determine the causes of sudden cardiac death in people aged 35 years or younger. DESIGN AND SETTING: A review of all autopsies performed between 1 January 1994 and 31 December 2002 at a major Sydney forensic medicine department serving an area with over 2 million people. MAIN OUTCOME MEASURES: Incidence of various types of cardiac disease causing sudden death in those aged 8-12 mm), and the granulosa layer of F3 follicles. The expression of leptin receptor in the granulosa layer of F2 and F1 follicles was barely detectable. This was in contrast to a much higher expression of leptin receptor maintained in the theca layer of F3-F1 follicles. The present results indicate that in chickens leptin might be involved in the adaptation to starvation due to attenuation of follicular apoptosis. The presence of leptin receptors in the ovary indicates the possibility of a peripheral effect of the hormone. PMID- 14748694 TI - Involvement of enzymes of amino acid metabolism and tricarboxylic acid cycle in bovine oocyte maturation in vitro. AB - Few studies demonstrate at a biochemical level the metabolic profile of both cumulus cells and the oocyte during maturation. The aim of the present study was to investigate the differential participation of enzymatic activity in cumulus cells and in the oocyte during in vitro maturation (IVM) by studying the activity of enzymes involved in the control of amino acid metabolism, alanine aminotransferase (ALT) and aspartate aminotransferase (AST); and the tricarboxylic acid (TCA) cycle, isocitrate dehydrogenase (IDH) and malate dehydrogenase (MDH). No NAD-dependent isocitrate dehydrogenase (NAD-IDH) activity was recorded in cumulus-oocyte complexes (COCs). ALT, AST, NADP-dependent isocitrate dehydrogenase (NADP-IDH) and MDH enzymatic units remained constant in cumulus cells and oocytes during IVM. Specific activities increased in oocytes and decreased in cumulus cells as a result of IVM (P<0.05). Similar activity of both transaminases was detected in cumulus cells, unlike in the oocyte, in which activity of AST was 4.4 times greater than that of ALT (P<0.05). High NADP-IDH and MDH activity was detected in the oocyte. Addition of alanine, aspartate, isocitrate + NADP, oxaloacetate or malate + NAD to maturation media increased the percentage of denuded oocytes reaching maturation (P<0.05), in contrast to COCs in which differences were not observed by addition of these substrates and co enzymes. The activity of studied enzymes and the use of oxidative substrates denotes a major participation of transaminations and the TCA cycle in the process of gamete maturation. The oocyte thus seems versatile in the use of several oxidative substrates depending on the redox state. PMID- 14748695 TI - Autologous and homologous transplantation of bovine spermatogonial stem cells. AB - The aim of this study was to develop a method for spermatogonial stem cell transplantation into the bovine testis. Five-month-old Holstein-Friesian calves were used and half of the calves were hemicastrated to allow autologous transplantation and the other half were used for homologous transplantation. Approximately 20 g of each testis was used for cell isolation. On average 106 cells per gram of testis containing about 70% type A spermatogonia were isolated. The cells were frozen in liquid nitrogen until transplantation. Testes were irradiated locally with 10-14 Gy of X-rays to deplete endogenous spermatogenesis. At 2 months after irradiation, cells (approximately 10 x 10(6) were injected into the rete testis through a long injection needle (18 gauge), using ultrasonography and an ultrasound contrast solution. At 2.5 months after transplantation, calves were castrated and samples of testes were taken for histological examination. After 2.5 months in the irradiated non-transplanted control testes, only 45% of the tubules contained type A spermatogonia. However, after autologous spermatogonial transplantation, >80% of the tubule cross-sections contained type A spermatogonia. In addition, only 20% of the tubules of the control testes contained spermatocytes and, except for a few tubules (5%) with round spermatids, no more advanced germ cells were found. After autologous spermatogonial transplantation, about 60% of the tubules contained spermatocytes; 30% contained spermatids and in about 15% of tubules spermatozoa were found. No improvement in spermatogonial repopulation was found after homologous transplantation. The results of this study demonstrate, for the first time, successful autologous transplantation of bovine spermatogonial stem cells resulting in a complete regeneration of spermatogenesis. PMID- 14748696 TI - Hormonal and behavioural changes during the mating season and pregnancy in Alpine marmots (Marmota marmota). AB - Under natural and artificial conditions, Alpine marmots (Marmota marmota) are true hibernators with a single breeding season starting immediately upon emergence from hibernation. Over three mating and breeding seasons, hormonal and mating patterns of colony-housed reproductive female marmots were investigated after exit from hibernation. Blood samples were taken for progesterone, oestrogen and relaxin assays with parallel ultrasound investigations. Copulations were observed from the first day after exit from hibernation until the end of pregnancy and reached a maximum number on day 37 before parturition. Mating behaviour was observed between the dominant animals as well as between dominant and subdominant group members. In the first week after exit from hibernation, plasma progesterone was detected in half of the animals. During the third week, progesterone concentrations were significantly higher in pregnant than in non pregnant animals or animals that had aborted. Immediately after emerging from hibernation, all successfully mated females showed higher serum relaxin values than non-successfully mated animals and this increase in relaxin concentration lasted until the end of pregnancy. The total concentration of oestrogen did not differ between pregnant and non-pregnant animals. The results of this study indicate that progesterone and relaxin might be useful indicators of early pregnancy in Alpine marmots. PMID- 14748697 TI - Fas and Fas ligand protein and mRNA in normal and atretic mouse ovarian follicles. AB - Apoptosis is the underlying mechanism of follicular atresia in the mammalian ovary. However, the apoptotic pathways governing this ovarian process are not completely elucidated. In the present study, expression of Fas and Fas ligand, the proximal members of the death receptor pathway, was evaluated in mouse ovarian follicles using immunofluorescence and in situ hybridization. Normal or atretic follicles were obtained from immature female Swiss mice after administration of 10 iu equine chorionic gonadotrophin for 48 or 72 h, respectively. Expression of both Fas and Fas ligand mRNA and protein was observed in granulosa cells of normal and atretic follicles. Although the oocytes of normal follicles failed to show any staining, those of atretic follicles stained intensely for Fas, indicating that the presence of Fas in the oocyte determines the fate of the follicle. PMID- 14748698 TI - Expression of Fas and Fas ligand protein and mRNA in mouse oocytes and embryos. AB - During mammalian embryonic development, abnormal eggs and embryos are eliminated by apoptosis; however, the precise apoptotic pathways remain as yet unidentified. In the present study, expression of Fas and Fas ligand - the proximal members of the death receptor pathway, was evaluated in mouse preimplantation embryos by immunofluorescence and in situ hybridization techniques. Ovulated oocytes were collected from oviducts of cyclic mice on the day of oestrus (day 0), and one cell, two-cell embryos and eight-cell morulae were collected from oviducts of mated animals on days 1, 2 and 3 of pregnancy, respectively. Blastocysts were flushed from the uterine horns on day 4. Expression of Fas and Fas L mRNAs and proteins was absent from embryos at days 0, 1 and 2. A marked increase in Fas and Fas L mRNA and protein expression was detected in all morphologically normal embryos on day 3 and day 4. In addition, embryos on days 3 and 4 were positive for terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling (TUNEL) staining; however, absence of caspase 8 and 3 and intense localization of proliferating cell nuclear antigen confirmed the proliferative status of these embryos. Furthermore, TUNEL staining was absent in postimplantation embryonic sections obtained on day 6. The results of the present studies thus indicate an equilibrium between proliferation and apoptosis in the preimplantation embryo. PMID- 14748699 TI - Effect of neonatal chronic stress on expression of Hsp70 and oestrogen receptor alpha in the rat oviduct during development and the oestrous cycle. AB - A chronic unpredictable stress model used to produce depressive disorders in adult rats was applied to neonatal rats to investigate whether this type of stress can induce changes in the expression of Hsp70 and oestrogen receptor alpha in the oviduct, as detected by immunohistochemistry. Rats stressed during neonatal development showed changes in the expression pattern of Hsp70. In neonatal control rats, Hsp70-positive cells observed in the isthmus did not show any changes. Moreover, rats exposed to this stress model that reached adulthood had higher expression of Hsp70 in the isthmus (P<0.01) but not in the ampulla during oestrus than did the control rats. In contrast, during dioestrus, no significant changes were noted in adult rats that were stressed during neonatal development or in rats that were stressed in adulthood. These findings indicate that the isthmus is very sensitive to stressful stimuli and that repeated pre weaning stress can change the expression of heat shock proteins in early and adult life. These subtle changes of expression in the oviduct did not affect the fertility of the rats that reached adulthood or that were mated under unstressed conditions. However, the control animals stressed during adulthood showed a disruption of the oestrous cycle: this finding is not observed in rats stressed during neonatal development that show an attenuated oestrous cycle disruption induced by chronic stress in adulthood. Moreover, there was dissociation between the expression of oestrogen receptor alpha and Hsp70. The amount of oestrogen receptor alpha remained constant in the epithelium of the oviduct in the control and in the stressed rats. Expression of oestrogen receptor alpha was noted in the stroma of the oviduct without the concomitant expression of Hsp70. It is possible that in certain cells and tissues Hsp70 is not necessary for oestrogen receptor alpha to be functional or Hsp70 might be present at very low amounts but is sufficient for the receptor to function. PMID- 14748700 TI - Overcoming poor in vitro nuclear maturation and developmental competence of domestic cat oocytes during the non-breeding season. AB - The domestic cat experiences circannual variations in ovarian activity and intrafollicular oocyte quality. One result is poor nuclear and cytoplasmic maturation during in vitro maturation (IVM) conducted during the annual non breeding season (July through November). In an attempt to overcome this seasonal phenomenon immature oocytes were collected from July through November and cultured in a conventional IVM medium (IVM1) or in IVM1 supplemented with different FSH concentrations and antioxidant (ascorbic acid or cysteine). Nuclear status of oocytes was assessed after IVM or IVF. Embryo stage and blastocyst quality were evaluated after 7 days of in vitro culture. Although the addition of antioxidant alone had no effect, the presence of 10 microg FSH ml(-1) improved nuclear maturation (75.4+/-4.1% versus 48.7+/-8.8% in IVM1; P<0.05) and fertilization success (47.9+/-3.2% versus 35.0+/-5.1% in IVM1; P<0.05). Furthermore, developmental competence of fertilized oocytes was enhanced (P<0.05) only in the presence of ascorbic acid (30.6+/-6.7%) or cysteine (33.6+/-5.1%) compared with IVM1 (8.1+/-8.8%). Consequently, blastocyst yield (17% of total oocytes cultured) was highest when oocytes were matured in medium containing higher FSH concentration and antioxidants. The results of this study demonstrate that meiotic and developmental competences are inherent to the immature cat oocyte collected during the non-breeding season. However, appropriate mechanisms (perhaps seasonal variation in FSH receptors or lack of antioxidant capacity of the cumulus-oocyte complex) are inadequate during this period of gonadal quiescence. Regardless, this compromised oocyte function during the non-breeding season can be overridden by altering in vitro culture conditions to include supplemental FSH and antioxidant. PMID- 14748701 TI - Mating-induced cumulus-oocyte maturation in the shrew, Suncus murinus. AB - The musk shrew, Suncus murinus, is an induced ovulator, the cumulus oophorus of which is unusual in several respects: it has no matrix, it always induces the acrosome reaction and it appears to be essential for fertilization. The present study documents distinctive features of the cumulus oophorus before and after ovulation, and of copulation-induced maturation of the ovulatory follicle, which has no antrum. In unmated females, potentially responsive ovarian follicles are distinguishable from large secondary follicles by differentiation of the granulosa into outer and inner cell layers, the latter being characterized particularly by intracellular glycogen deposits. The average number of responsive follicles equates with the number that ovulate. By about 10 h after mating, meiosis has reached metaphase II, with extrusion of the first polar body. Coincidentally, a cavity has developed between the inner and outer follicular layers, demarcating the smaller cells of the granulosa from the glycogen-rich cells of the cumulus oophorus. Subsequently, the glycogen becomes restricted primarily to the inner cumulus, and the corona cells began to retreat from the zona pellucida surface to form an unusual very distinct perizonal space that is clearly evident at the time of ovulation. The cumulus is stabilized by gap and tight junctions, and presents a smooth external surface that appears to initiate the acrosome reaction. After fertilization, at which time the zona pellucida becomes more resistant to both trypsin and dithiothreitol, the cumulus develops intercellular lacunae, and is eventually discarded about 15 h after ovulation. PMID- 14748702 TI - Linking endocytic cargo to clathrin: structural and functional insights into coated vesicle formation. AB - Clathrin-mediated endocytosis is the major process by which transmembrane proteins are internalized from the cell's limiting membrane into the first compartment of the endosomal system, the early endosome. From here, these transmembrane cargo proteins, which are of widely varying type and function, are trafficked to their required destination. Endocytosis plays, therefore, an important role in cell signalling, nutrient uptake, cellular homoeostasis and the interaction of the cell with its external environment. The formation of clathrin coated endocytic vesicles requires the complex interplay of many proteins with each other and with the membrane itself. Their formation has served as a paradigm for formation of all types of transport vesicle, which move cargo between the various membrane-bound compartments of the cell. Clathrin-coated vesicles (CCVs) possess three layers: the inner membrane layer, in which the transmembrane cargo is embedded, linked to the outer clathrin lattice by a layer of cargo-binding adaptors and proteins that aid and regulate vesicle formation. Protein X-ray crystallography in combination with biochemical, biophysical and cell biological assays has been used to investigate the structure and function of some of the proteins that make up the middle layer of CCVs. These proteins are diverse in their functions, but are all modular in nature, consisting of folded domains joined by long unstructured linkers. Within these linkers are short motifs that interact with the folded domains of other components of the CCV formation machinery. Many of these folded domains also bind directly to the membrane. These interactions, whose molecular basis we have studied, have affinities in the low micromolar range, making them readily reversible and easily regulated. The mechanism of CCV formation is discussed in the light of this structural and biochemical data. PMID- 14748703 TI - General considerations for proteolytic cascades. AB - Proteases are involved in the regulation of a wide variety of essential physiological processes, often by participating in a highly orchestrated sequence of events termed a 'proteolytic cascade'. Four major proteolytic cascades with disease relevance are candidates for therapeutic intervention, namely caspase mediated apoptosis, blood coagulation, the matrix metalloproteinase cascade and the complement cascade. Understanding the various steps involved in the functioning of a cascade is key to deciding possible points of intervention for the design of potential drug molecules. This brief review illustrates some of the common features of proteolytic cascades using the blood coagulation pathway as an example. PMID- 14748704 TI - Matrix metalloprotease inhibitors: design from structure. AB - The zinc- and calcium-dependent family of proteins called the MMPs (matrix metalloproteases) are collectively responsible for the degradation of the extracellular matrix. The enzymes are synthesized as zymogens, and under physiological conditions are selectively regulated by endogenous inhibitors. An imbalance between the active enzymes and their natural inhibitors leads to the accelerated destruction of connective tissue associated with the pathology of diseases such as arthritis, cancer, multiple sclerosis and cardiovascular diseases. The potential for using specific enzyme inhibitors as therapeutic agents to redress this balance has led to intensive research focused on the design, synthesis and molecular deciphering of low-molecular-mass inhibitors of this family of proteins. The design of early MMP inhibitors was based on the scissile site sequence of peptide substrates, with moieties customized to chelate the critical zinc ion at the enzymes' active site. These initial efforts were supported by X-ray and NMR data on MMP complexes, exploiting sequence and structural differences in the principal specificity pocket of the enzymes, leading to subtype-selective MMP inhibitors. This review will provide a critical appraisal of the design principles that have been utilized in generating molecules that inhibit MMPs, and explore issues relevant to obtaining clinical efficacy of MMP inhibitor-based therapies. PMID- 14748705 TI - Proteases of the complement system. AB - The complement system is a group of about 35 soluble and cell-surface proteins which interact to recognize, opsonize and clear or kill invading micro-organisms or altered host cells (e.g. apoptotic or necrotic cells). Complement is a major part of the innate immune system. Recognition proteins such as C1q, MBL (mannan binding lectin) and ficolins bind to targets via charge or sugar arrays. Binding causes activation of a series of serine protease proenzymes, such as C1r, C1s and MASP2 (MBL-associated serine protease 2), which in turn activate the atypical serine proteases factor B and C2, which then activate the major opsonin of the system, C3. Activated C3 binds covalently to targets, and is recognized by receptors on phagocytic cells. Two of the complement proteases, factors D and I, circulate not as proenzymes, but in activated form, and they have no natural inhibitors; their substrates are transient protein complexes (e.g. C3bB and C3bH) which form during complement activation. Factor B and C2 also have no natural inhibitor; they are active only when proteolytically cleaved and bound in an unstable, short-lived complex with C3b or C4b. C1r, C1s and the MASPs, in contrast, are regulated more conventionally by the natural serpin, C1-inhibitor. Complement proteases in general have very narrow specificity, and low substrate turnover with both natural and synthetic substrates. Excessive activation of complement is inflammatory, and causes tissue damage (e.g. in rheumatoid arthritis, or in ischaemia/reperfusion injury). Substances that regulate complement activation are likely to be useful in the regulation of inflammation. Complement activation might potentially be controlled at many different steps. Much attention has been focused on controlling the formation or activity of the protease complexes C3bBb and C4b2a (containing activated factor B and C2 respectively), as these generate the inflammatory peptides C3a and C5a. PMID- 14748706 TI - Improvement of the anti-C3 activity of compstatin using rational and combinatorial approaches. AB - Compstatin is a 13-residue cyclic peptide that has the ability to inhibit the cleavage of C3 to C3a and C3b. The effects of targeting C3 cleavage are threefold, and result in hindrance of: (i) the generation of the pro-inflammatory peptide C3a, (ii) the generation of opsonin C3b (or its fragment C3d), and (iii) further complement activation of the common pathway (beyond C3) with the end result of the generation of the membrane attack complex. We will report on our progress on: (i) rational design of more active compstatin analogues based on the three-dimensional structure of compstatin, (ii) experimental combinatorial design based on the generation of a phage-displayed peptide library partially randomized with the implementation of structure-induced restraints, and (iii) theoretical combinatorial design based on a novel computational optimization method, structure-induced restraints and flexible structural templates. All three approaches have resulted in analogues with improved activities. Currently, the lead analogue has the sequence acetyl-I[CVYQDWGAHRC]T-NH(2) (where the brackets denote cyclization), and is 16-fold more active than the parent peptide. We will also report on our progress towards understanding the dynamic character of compstatin using molecular dynamics simulations. The identification of an ensemble of interconverting conformers of compstatin with variable populations is a first step towards the incorporation of dynamic elements in the design of new analogues using dynamics-activity relationships in addition to structure-activity relationships. PMID- 14748707 TI - Proteolytic cascade in the amyloidogenesis of Alzheimer's disease. AB - beta-Amyloid, a neurotoxic peptide deposited in the brains of Alzheimer's disease patients, is released by a series of membrane-limited proteolytic events. beta Secretase activity is enhanced by cellular targeting into intracellular cholesterol-rich microdomains, which are dispersed by statins. PMID- 14748708 TI - Improved RNA cleavage by LNAzyme derivatives of DNAzymes. AB - Specific cleavage of RNA is catalysed by short oligodeoxynucleotides termed DNAzymes. DNAzymes consist of two binding arms that hybridize to a predetermined RNA sequence and a catalytic core that cleaves a phosphodiester bond held between the binding arms. DNAzymes are exemplified by the well-studied 10-23 DNAzyme, which compared with protein ribonucleases is highly specific, albeit slow. Here we report a significant improvement in cleavage kinetics, while maintaining specificity, by incorporation of LNA (locked nucleic acid) and alpha-L-LNA nucleotides into the binding arms of 10-23 DNAzyme. DNAzymes modified in this way (LNAzymes) enhance cleavage of a phosphodiester bond presented in a short RNA substrate as well as in longer and highly structured substrates, and efficient cleavage is maintained from single- to multiple-turnover conditions. Analysis of the cleavage reaction indicates that substrate hybridization is boosted by the presence of the locked residues within the LNAzymes, while no apparent change occurs in the catalytic strand-scission step. PMID- 14748709 TI - Single-molecule studies of DNA and RNA four-way junctions. AB - Branched helical junctions are common in nucleic acids. In DNA, the four-way junction (Holliday junction) is an essential intermediate in homologous recombination and is a highly dynamic structure, capable of stacking conformer transitions and branch migration. Our single-molecule fluorescence studies provide unique insight into the energy landscape of Holliday junctions by visualizing these processes directly. In the hairpin ribozyme, an RNA four-way junction is an important structural element that enhances active-site formation by several orders of magnitude. Our single-molecule studies suggest a plausible mechanism for how the junction achieves this remarkable feat; the structural dynamics of the four-way junction bring about frequent contacts between the loops that are needed to form the active site. The most definitive evidence for this is the observation of three-state folding in single-hairpin ribozymes, the intermediate state of which is populated due to the intrinsic properties of the junction. PMID- 14748710 TI - In vitro and in vivo effects of oxidative damage to deoxyguanosine. AB - Biochemical, biophysical and biological studies of oligonucleotides containing lesions at defined sites provide a molecular basis for the effects of DNA lesions. dG (deoxyguanosine) is the most easily oxidized of the four native nucleotides. The chemical reactivity of dG correlates with compilations of mutations, which reveal that a significant fraction of transitions or transversions involve dG. OxodG (7,8-dihydro-8-hydroxy-2'-deoxyguanosine) is widely recognized as an important lesion derived from the oxidation of dG, and significant effort has been expended in studies of its effects on DNA structure and function. Recently, the properties of other lesions derived from dG and/or the oxidation of OxodG have been uncovered. Studies on these lesions reveal that they too are biologically significant. PMID- 14748711 TI - Semi-synthetic DNA-protein conjugates: novel tools in analytics and nanobiotechnology. AB - This article reports on the syntheses, characterization and applications of semi synthetic conjugates composed of nucleic acids, proteins and inorganic nanoparticles. For example, self-assembled oligomeric networks consisting of streptavidin and double-stranded DNA are applicable as reagents in immunoassays, model systems for ion-switchable nanoparticle networks as well as nanometer scaled 'soft material' standards for scanning probe microscopy. Covalent conjugates of single-stranded DNA and streptavidin are utilized as biomolecular adapters for the immobilization of biotinylated macromolecules at solid substrates via nucleic acid hybridization. This 'DNA-directed immobilization' allows for reversible and site-selective functionalization of solid substrates with metal and semiconductor nanoparticles or, vice versa, for the DNA-directed functionalization of gold nanoparticles with proteins, such as immunoglobulins and enzymes. This approach is applicable for the detection of chip-immobilized antigens. Moreover, covalent DNA-protein conjugates allow for their selective positioning along single-stranded nucleic acids, and thus for the construction of nanometre-scale assemblies composed of proteins and/or nanoclusters. Examples include the fabrication of functional biometallic nanostructures from gold nanoparticles and antibodies, applicable as diagnostic tools in bioanalytics. PMID- 14748712 TI - Dietary fatty acids and chylomicron synthesis and secretion. AB - There is currently considerable interest in potential atherogenic and thrombogenic consequences of elevated concentrations of triacylglycerols, especially in the post-prandial state. Despite this, there is limited information on the effects of dietary fatty acids on the synthesis, secretion and metabolism of chylomicrons, the large triacylglycerol-rich lipoproteins synthesized in the enterocyte following the digestion and absorption of dietary fat. This brief review considers current approaches to the investigation of chylomicron synthesis and summarizes some of the human, cell and animal studies that have investigated effects of different fatty acids on these pathways. Potential sites for modulatory effects of dietary fatty acids on the molecular events of chylomicron synthesis are proposed in the light of the recent model that has been developed from cell and animal studies and observations based on abnormalities in chylomicron formation in human inherited autosomal recessive diseases. PMID- 14748713 TI - Synthesis and function of hepatic very-low-density lipoprotein. AB - Most of the triacylglycerol (TAG) utilized for the assembly of very-low-density lipoprotein (VLDL) in the secretory apparatus of the hepatocyte is mobilized by lipolysis of the cytosolic TAG pool, followed by re-esterification. The lipases involved include arylacetamide deacetylase and/or triacylglycerol hydrolase. Some of the re-esterified products of lipolysis gain access to an apolipoprotein-B rich VLDL precursor to form mature VLDL. Some, however, are returned to the cytosolic pool in a process that is stimulated by insulin and inhibited by microsomal triacylglycerol transfer protein (MTP). Phospholipids also contribute to VLDL TAG in a process which involves ADP-ribosylation factor-1 (ARF-1) mediated activation of phospholipase D. The temporary storage of TAG in the liver, followed by its mobilization and secretion as VLDL, form part of a process by which the liver protects vulnerable body tissues from excess lipotoxic non esterified ('free') fatty acids in the plasma. PMID- 14748714 TI - Membrane cholesterol and the regulation of signal transduction. AB - The plasma membrane of mammalian cells consists of microdomains differing in lipid and protein composition. Two distinct classes of cholesterol/sphingolipid microdomain (caveolae and lipid rafts) are assembly points for transmembrane signalling complexes. Recent evidence suggests that transient changes in cholesterol content may be important in regulating signal transduction. PMID- 14748715 TI - The genes and proteins of atherogenic lipoprotein production. AB - Dietary fat provides a major source of nutrition, but may in excess lead to obesity, insulin resistance, high blood cholesterol levels and atherosclerosis. Here we report molecular events that co-ordinate whole-body lipid homoeostasis from insects to humans, viewed in the context of rare and common genetic disorders of apolipoprotein B-containing lipoprotein production. PMID- 14748716 TI - Enterocyte fatty acid uptake and intestinal fatty acid-binding protein. AB - This article reviews our current understanding of the uptake of fatty acids by the enterocytes of the intestine. The micellar solubilization of fatty acids by bile salts and the factors regulating that process are discussed. The mechanism of how micellar solubilization of fatty acids promotes the uptake of fatty acids by enterocytes and their relative importance is reviewed. Additionally, discussion of the various fatty acid transporters located at the brush border membrane of the enterocytes is included. Finally, a summary of our current understanding of the function of fatty-acid-binding proteins inside enterocytes is provided. PMID- 14748717 TI - Chylomicron metabolism. AB - Chylomicrons are the 'orphans' of the lipoprotein family. Difficulty of measurement has impeded understanding of their metabolism. Plasma concentrations of chylomicrons and chylomicron remnants give no insight into the magnitude of substrate flux through these pathways. A defect in clearance of chylomicron remnants is probably an indication of a more generalized defect in lipoprotein metabolism. Accumulating evidence supports a relationship between abnormalities in the clearance from plasma of chylomicron remnants and accelerated progression of atherosclerosis. Methods using stable isotopes in appropriately formulated emulsions are providing valuable new information. PMID- 14748718 TI - Increased FAT (fatty acid translocase)/CD36-mediated long-chain fatty acid uptake in cardiac myocytes from obese Zucker rats. AB - Disturbed cardiac lipid homoeostasis in obesity is regarded as a key player in the development of cardiovascular diseases. In this study, we show that FAT (fatty acid translocase)/CD36-mediated LCFA (long-chain fatty acid) uptake in cardiac myocytes from young adult obese Zucker rats is markedly increased, but insensitive to insulin. Basal and insulin-induced glucose uptake rates in these myocytes are not changed, suggesting that during the development from obesity to hyperglycaemic Type II diabetes, alterations in cardiac LCFA uptake precede alterations in cardiac glucose uptake. PMID- 14748719 TI - The surprising diversity of Delta6-desaturase substrates. AB - A single gene encoding a Delta6-desaturase (FADS2) has been isolated and characterized in mammalian species. This Delta6-desaturase plays a major role in the biosynthesis of PUFAs (polyunsaturated fatty acids). It catalyses the rate limiting desaturation of linoleic acid (C(18:2) n-6) and alpha-linolenic acid (C(18:3) n-3) required for the biosynthesis of long-chain PUFAs. Moreover, recent studies have provided strong evidence that this Delta6-desaturase also acts on 24 carbon PUFAs of both the n-6 and n-3 series. Another substrate of this Delta6 desaturase has been identified through complementary works from different investigators. This Delta6-desaturase acts on a saturated fatty acid, palmitic acid (C(16:0)), leading to the newly characterized biosynthesis of hexadecenoic acid (C(16:1) n-10) or sapienate. PMID- 14748720 TI - Regulation of lipoprotein trafficking in the kidney: role of inflammatory mediators and transcription factors. AB - Inflammation and dyslipidaemia both play important roles in the development of glomerular atherosclerosis in renal diseases. We have demonstrated that inflammatory mediators induced Scr (scavenger receptor) expression and the formation of foam cells, and that AP-1 (activator protein 1)/ets were necessary transcriptional factors for Scr induction in HMCs (human kidney mesangial cells). Most cells are protected from excessive native LDL (low-density lipoprotein) accumulation by tight feedback regulation of the LDLr (LDL receptor). However, we observed that HMCs formed foam cells via the LDLr pathway when incubated with IL 1beta (interleukin-1beta; 5 ng/ml) and unmodified LDL (200 microg/ml), suggesting that inflammatory mediators may disrupt the cholesterol-mediated feedback regulation. This feedback involves cholesterol-mediated down-regulation of LDLr controlled by SCAP [SREBP (sterol responsive element-binding protein) cleavage activating protein]. We have also demonstrated that both tumour necrosis factor alpha and IL-1beta increased nuclear SREBP-1 levels by increasing SCAP mRNA expression, even in the presence of a high concentration of LDL. Since intracellular lipid content is governed by both influx and efflux mechanisms, we set out to examine the impact of inflammatory cytokines on cholesterol efflux, a process mediated by the protein ABCA1 (ATP binding cassette A1). IL-1beta inhibited [(3)H]cholesterol efflux from HMCs by inhibition of the peroxisome proliferator-activated receptor/LXR (liver X receptor)/ABCA1 pathway. Taken together, our results suggest that inflammatory mediators increase lipid accumulation in HMCs not only by promoting increased lipoprotein uptake by Scr and LDLr, but also by inhibiting ABCA1-mediated cholesterol efflux to high density lipoprotein. PMID- 14748721 TI - LXR (liver X receptor) and HNF-4 (hepatocyte nuclear factor-4): key regulators in reverse cholesterol transport. AB - Cholesterol homoeostasis is the result of the fine tuning between intake and disposal of this molecule. High levels of cholesterol in the blood are detrimental as they may lead to excessive accumulation in vessel walls, a condition predisposing to the development of atherosclerotic lesions. Cholesterol is removed from the vessel wall and transported to the liver through a process called reverse cholesterol transport. Nuclear receptors are among the most important transcription factors regulating genes involved in different steps of reverse cholesterol transport. Here, we discuss the role of the nuclear receptors LXR (liver X receptor) and HNF-4alpha (hepatocyte nuclear factor-4alpha) in different steps of reverse cholesterol transport. LXR controls the transcription of crucial genes in cholesterol efflux from macrophages and its transport to the liver, such as ABCA1 (ATP binding cassette A1), CYP27A1 (sterol 27-hydroxylase), CLA-1 (scavenger receptor type B1) and apolipoprotein E. Some oxysterols present in oxidized low-density lipoproteins and proinflammatory cytokines modulate the activity of LXR by antagonizing the effect of activators of this receptor, thus contributing to cholesterol accumulation in macrophages. Bile acid synthesis, which represents the final step of reverse cholesterol transport, is transcriptionally regulated by several nuclear receptors at the level of the liver-specific cytochrome P450 cholesterol 7alpha-hydroxylase (CYP7A1), the rate limiting enzyme of this metabolic pathway. Bile acids returning to the liver through the enterohepatic circulation down-regulate CYP7A1 transcription via the bile acid sensors farnesoid X receptor and HNF-4alpha. Based on this evidence, these nuclear receptors are candidate targets of new drugs for the treatment and prevention of atherosclerotic disease. PMID- 14748722 TI - Critical roles of the nuclear receptor PPARbeta (peroxisome-proliferator activated receptor beta) in skin wound healing. AB - The PPARs (peroxisome-proliferator-activated receptors) alpha, beta/delta and gamma belong to the nuclear hormone receptor superfamily. While all three receptors are undetectable in adult mouse interfollicular epidermis, PPARbeta expression and activity is strongly re-activated by inflammatory stimuli during epidermal injury. The pro-inflammatory cytokine TNFalpha (tumour necrosis factor alpha) stimulates transcription of the PPARbeta gene via an activator protein-1 site in its promoter and it also triggers the production of PPARbeta ligands in keratinocytes. This increase of PPARbeta activity in these cells up-regulates the expression of integrin-linked kinase and 3-phosphoinositide-dependent kinase-1, which phosphorylates protein kinase B-alpha (Akt1). The resulting increase in Akt1 activity suppresses apoptosis and ensures the presence of a sufficient number of viable keratinocytes at the wound margin for re-epithelialization. Together, these observations reveal that PPARbeta takes on multiple roles and contributes favourably to the process of wound closure. PMID- 14748723 TI - Adipocyte cholesterol balance in obesity. AB - Adipose tissue is specialized in the storage of energy in the form of triacylglycerol. Within the fat cell, triacylglycerols are found in a well defined structural compartment called the lipid droplet, which occupies the vast majority of the fat cell volume. However, many other lipids are present in the lipid droplet. These include sterols, carotenoids, cholecalciferol and lipophilic toxic pollutants of the environment such as dioxins and tocopherols. The topic of this article is the role of fat cell cholesterol in adipose tissue physiology and its potential implication in pathological states such as obesity. PMID- 14748724 TI - Transcriptional regulation of human SREBP-1c (sterol-regulatory-element-binding protein-1c): a key regulator of lipogenesis. AB - Sterol-regulatory-element-binding protein 1c (SREBP-1c) is one member of the family of transcription factors that stimulate sterol and fatty-acid biosynthesis in animal cells. Human SREBP-1c, mapped to chromosome 17p11.2, is expressed in liver, intestine, skeletal muscle and adipocytes. A section of genomic sequence from a chromosome 17 library, thought to contain the SREBP-1c promoter, was cloned. Putative transcription-factor-binding sites and a potential transcriptional start site were identified using the Genomatix Suite of sequence analysis tools (MatInspector). Sequence analysis showed the human promoter to be 42% identical with the previously published mouse sequence. Two novel transcription-factor-binding sites were identified: those for PDX-1 (pancreatic duodenal homoeobox-1) and HNF-4 (hepatic nuclear factor-4). Co-transfection experiments with overexpression plasmids for PDX-1 and HNF-4 suggested that both factors stimulate SREBP-1c gene expression, although further work is required to ascertain their mechanisms of action. PMID- 14748725 TI - Chylomicron-remnant-like particles modify production of vasoactive mediators by endothelial cells. AB - Endothelial-cell dysfunction is a critical initiating event in the pathogenesis of atherosclerosis. Although there is evidence to suggest that chylomicron remnants (CMRs), lipoproteins derived from the diet, influence endothelial-cell function to generate a pro-atherogenic phenotype, the mechanisms involved remain undefined. We have examined the effects of CMR-like particles (CMR-LPs) on human endothelial-cell function, focusing on the cyclo-oxygenase (COX) and nitric oxide synthase (NOS) pathways. CMR-LPs strongly enhanced the expression of the inducible cyclo-oxygenase COX-2 and increased prostacyclin synthesis in a biphasic manner. Studies with the COX-2-selective inhibitor NS-398 confirmed the COX-2 dependency of the later increase in prostanoid production. Pre-incubation with CMR-LPs reduced basal and thrombin-stimulated cGMP generation, whereas expression of endothelial NOS was not modified by remnant treatment. PMID- 14748726 TI - B-cell antigen receptor activates transcription factors NFAT (nuclear factor of activated T-cells) and NF-kappaB (nuclear factor kappaB) via a mechanism that involves diacylglycerol. AB - Engagement of the B-cell antigen receptor (BCR) induces the activation of various transcription factors, including NFAT (nuclear factor of activated T-cells) and NF-kappaB (nuclear factor kappaB), which participate in long-term biological responses such as proliferation, survival and differentiation of B-lymphocytes. We addressed the biochemical basis of this process using the DT40 chicken B-cell lymphoma. We discovered that Bruton's tyrosine kinase (BTK) and phospholipase C gamma2 (PLC-gamma2) are required to activate NFAT and NF-kappaB, and to produce the lipid second messenger diacylglycerol in response to BCR cross-linking. Therefore the functional integrity of the BTK/PLC-gamma2/diacylglycerol signalling axis is crucial for BCR-directed activation of both transcription factors NFAT and NF-kappaB. PMID- 14748727 TI - Scavenger receptor class B type I in high-density lipoprotein metabolism, atherosclerosis and heart disease: lessons from gene-targeted mice. AB - The scavenger receptor class B type I (SR-BI) is a multi-ligand receptor that can mediate the binding and bi-directional lipid transfer between high-density lipoproteins (HDLs) and cells. It is expressed in a variety of tissues, including the liver, and in macrophages in atherosclerotic plaques. The physiological role of SR-BI has been tested in vivo by the genetic manipulation of SR-BI levels in mice. Mice lacking SR-BI exhibit impaired hepatic-selective HDL cholesterol uptake and increased atherosclerosis, suggesting that SR-BI is required for hepatic reverse cholesterol transport and normally protects against atherosclerosis. Surprisingly, elimination of SR-BI in apolipoprotein E knockout mice results in rapid development of occlusive coronary artery disease, accompanied by spontaneous myocardial infarction, reduced heart function and early death, which points to a role for SR-BI in protection against coronary heart disease. The in vivo role of macrophage SR-BI has been less clear. We have used bone-marrow transplantation to demonstrate that bone-marrow-derived SR-BI also normally protects against atherosclerosis in low-density lipoprotein receptor knockout mice. These results suggest that SR-BI may have multiple protective effects against atherosclerosis in different tissues. PMID- 14748728 TI - Caveolins and membrane cholesterol. AB - Caveolae (small plasma membrane invaginations) and their coat proteins, caveolins, have attracted the attention of researchers in diverse fields, including cell biology, cardiovascular and cancer research. The tight association between caveolin and cholesterol governs the biochemical behaviour of caveolae and is emerging as an important characteristic in a number of processes assigned to these multifunctional organelles. In this review, selected aspects of the caveolin-cholesterol association and its potential functional implications are discussed. PMID- 14748729 TI - ATP-binding cassette transporter A1: regulation of cholesterol efflux. AB - The ATP-binding cassette transporter A1 (ABCA1) is involved in the regulation of cholesterol efflux from cells. Mutations in ABCA1 give rise to familial high density lipoprotein (HDL) deficiency and Tangier disease, which is characterized by very low levels of HDL in plasma and cholesteryl ester accumulation in tonsils and other reticuloendothelial cells. The mechanism of action of ABCA1 is still unclear, but requires the transfer of phospholipid and cholesterol to apolipoprotein A1 bound by or close to the transporter. An important factor in the regulation of ABCA1 is cholesterol itself, which provides oxysterol ligands for liver X receptors that stimulate ABCA1 transcription. ABCA1-deficient mice show increased cholesterol absorption, suggesting that ABCA1 could also help to transport dietary cholesterol back out of intestinal absorptive cells into the lumen. Thus ABCA1 is intimately connected to various aspects of the regulation of whole-body cholesterol metabolism and probably plays an important role in protecting against the development of cardiovascular disease. PMID- 14748730 TI - Circulating levels of the chemokines JE and KC in female C3H apolipoprotein-E deficient and C57BL apolipoprotein-E-deficient mice as potential markers of atherosclerosis development. AB - We have investigated serum chemokines for their suitability as markers of atherosclerosis development in apoE (apolipoprotein E)-deficient ((-/-)) mice. Female C3H apoE(-/-) and C57BL apoE(-/-) mice were fed on either diet W (Western diet; 6 weeks) or normal rodent diet (12 weeks). Serum lipids (0, 6 and 12 weeks) and terminal chemokine levels were measured using commercially available assays, whereas the lesion area was determined using Oil-Red O-stained aortic sections. Serum lipids were higher in C3H apoE(-/-) mice for both diets throughout the study; however, lesions were significantly larger in C57BL apoE(-/-) mice fed on either diet. Chemokine levels were significantly lower in C3H apoE(-/-) mice fed on the normal diet, but no difference was observed between the two groups fed on diet W. We conclude that serum chemokine levels are potential markers for atherosclerosis susceptibility in C3H and C57BL apoE(-/-) mice fed on a normal rodent diet. PMID- 14748731 TI - Faecal sterol output is increased by arachidyl amido cholanoic acid (Aramchol) in rats. AB - Fatty acid-bile acid conjugates (FABACs) were shown recently to have important and multiple effects on cholesterol metabolism. In human fibroblasts, they were found to markedly enhance cholesterol efflux by an ATP-binding cassette transporter A1-dependent pathway. In C57L/J mice, they increased CYP7A1 activity and RNA expression, while decreasing moderately 3-hydroxy-3-methylglutaryl-CoA reductase activity. In C57L/J mice and in hamsters, they also decreased serum cholesterol levels, whereas in other animals, this effect was not seen in short term experiments. In the present study, we investigated potential mechanisms of action of arachidyl amido cholanoic acid (Aramchol), with particular reference to biliary and faecal sterol outputs in rats. Supplementation with Aramchol at a dose of 150 mg x kg(-1) x day(-1) increased neutral sterol output by approx. 50%, while the faecal outputs of bile salts and total sterols increased by almost 2 fold. Biliary lipid outputs were not significantly different between the control and FABAC-supplemented animals. These findings indicate an overall catabolic effect of FABACs on body cholesterol. PMID- 14748732 TI - Lipid oxidation in atherogenesis: an overview. AB - The 'oxidation theory' for atherosclerosis proposes that lipid and/or protein oxidation products are responsible for lesion formation/development. The major target for oxidation is suggested to be intimal low-density lipoprotein. This idea was stimulated by the pro-atherogenic properties of in vitro oxidized lipoproteins, such as stimulation of chemotaxis and sterol accumulation in macrophages, adhesion molecule expression on endothelial cells and apoptosis of several cell types. It was supported by detection of oxidation products in lesion lipoproteins, although these are (in general) less heavily oxidized and their biological activity is less rigorously defined than for their in vitro oxidized counterparts. Lesion lipids contain products generated by both enzymic and non enzymic oxidation reactions; the majority are generated non-enzymically. The type and source of oxidant involved has been the subject of much speculation and is not resolved. The oxidation theory predicts that appropriate antioxidants will protect against atherosclerosis. Vitamin E has been used in several animal and human studies, but to date has shown little evidence of anti-atherosclerotic potential. However, lack of knowledge of the oxidant(s) driving lesion oxidation and the complexity of the anti- and pro-oxidant properties of vitamin E may explain its disappointing track record to date. These subjects require more rigorous study before the oxidation theory can be fairly tested. PMID- 14748733 TI - Specific markers of lipid peroxidation issued from n-3 and n-6 fatty acids. AB - Several markers of lipid peroxidation are available with different degrees of specificity, from malondialdehyde as a global marker, to F(2)-isoprostane, which is specifically produced from arachidonic acid. Among these, 4-hydroxynonenal is recognized as a breakdown product of fatty acid hydroperoxides, such as 15 hydroperoxy-eicosatetraenoic acid and 13-hydroperoxy-octade cadienoic acid from the n -6 fatty acids. Furthermore, 4-hydroxyhexenal (4-HHE) derives from n -3 fatty acid hydroperoxides. We have recently described the occurrence of 4 hydroxydodecadienal (4-HDDE) from the 12-lipoxygenase product of arachidonic acid 12-hydroperoxy-eicosatetraenoic acid. These three hydroxy-alkenals may be measured in human plasma by GC-MS, but they may partly be generated in the course of sampling, and the relative volatility of 4-HHE makes its measurement quite unreliable. We have successfully characterized and measured the stable oxidized carboxylic acid products from the hydroxy-alkenals 4-HNA, 4-HHA and 4-HDDA in urine. The ratio between 4-HHA and 4-HNA found in the same urinary sample might provide useful information on the location of lipid peroxidation, accounting for the high enrichment of the cerebrovascular system with docosahexaenoic acid, the main n -3 fatty acid in humans. PMID- 14748734 TI - A novel antioxidant action of ethanolamine plasmalogens in lowering the oxidizability of membranes. AB - We have demonstrated a novel antioxidant action of ethanolamine plasmalogens both in protecting cholesterol from oxidation by free radicals and in lowering the oxidizability of membranes, along with the action of scavenging radicals, by the oxygen-uptake method using large unilamellar vesicles and the water-soluble azo radical initiator, AAPH [2,2'-azobis-(2-amidino-propane) dihydrochloride]. PMID- 14748735 TI - Connection of lipid peroxide oxidation with the sphingomyelin pathway in the development of Alzheimer's disease. AB - Alzheimer's disease (AD) is characterized by progressive decline in cognition, memory and intellect. It has been hypothesized that amyloid-beta peptide (A-beta) may have a prominent role in neurodegeneration. Oxidative mechanisms have been implicated in this pathway. There is substantial evidence that inflammatory mechanisms, induced by tumour necrosis factor alpha (TNF-alpha), are also involved in AD. TNF-alpha activates receptors linked to multiple effector systems, including a sphingomyelin pathway and peroxide oxidation. We have determined the changes of neutral sphingomyelinase activity, sphingomyelin and ceramide contents, and the level of lipid peroxide products (conjugated dienes), in the cerebral cortex, hippocampus and cerebellum of rats within 3 h and 7 days of intracerebral injection of A-beta and TNF-alpha. A single injection of A-beta and TNF-alpha has been shown to increase the level of peroxide products in the hippocampus and cerebral cortex within 3 h and 7 days. Sphingomyelinase activity and ceramide levels have been found to increase 7 days after A-beta administration. We found that activation of the sphingomyelin pathway lies downstream from the oxidative stress. PMID- 14748736 TI - Plasmalogens: targets for oxidants and major lipophilic antioxidants. AB - Cellular membranes and plasma lipoproteins are less efficiently protected against oxidative stress than the various aqueous compartments of mammalian organisms. Here, previous results on the role of plasmalogens in lipid oxidation are evaluated on the basis of criteria required for an antioxidant. The plasmalogen specific enol ether double bond is targeted by a vast variety of oxidants, including peroxyl radicals, metal ions, singlet oxygen and halogenating species. Oxidation of the vinyl ether markedly prevents the oxidation of highly polyunsaturated fatty acids, and products of plasmalogen degradation do not propagate lipid oxidation. This protection is also demonstrated intramolecularly, thus ascertaining the function of plasmalogens as a major storage pool for polyunsaturated fatty acids. Although cells rapidly incorporate and synthesize plasmalogens de novo, their plasmalogen contents can be deliberately increased by supplementation with precursors. Thus plasmalogens terminate lipid-oxidation processes, are present in adequate locations at sufficient concentrations, and are rapidly regenerated, classifying them as efficient antioxidants in vitro. Future work should address the in vivo role of plasmalogens in lipid oxidation and the biological function of plasmalogen interactions with oxidants. PMID- 14748737 TI - Mechanisms of signal transduction mediated by oxidized lipids: the role of the electrophile-responsive proteome. AB - Cellular redox signalling is mediated by the post-translational modification of proteins by reactive oxygen/nitrogen species or the products derived from their reactions. In the case of oxidized lipids, several receptor-dependent and independent mechanisms are now emerging. At low concentrations, adaptation to oxidative stress in the vasculature appears to be mediated by induction of antioxidant defences, including the synthesis of the intracellular antioxidant glutathione. At high concentrations apoptosis occurs through mechanisms that have yet to be defined in detail. Recent studies have revealed a mechanism through which electrophilic lipids, formed as the reaction products of oxidation, orchestrate these adaptive responses in the vasculature. Using a proteomics approach, we have identified a subset of proteins in cells that we term the electrophile-responsive proteome. Electrophilic modification of thiol groups in these proteins can initiate cell signalling events through the transcriptional activation of genes regulated by consensus sequences for the antioxidant response element found in their promoter regions. The insights gained from our understanding of the biology of these mechanisms will be discussed in the context of cardiovascular disease. PMID- 14748738 TI - Antioxidants and atherosclerosis. AB - The oxidative modification of low-density lipoprotein (LDL) can be induced by various active species by different mechanisms. Vitamin E and other radical scavenging antioxidants can inhibit the free radical-mediated oxidation of LDL, but they are not effective against LDL oxidation induced by non-radical mechanisms. PMID- 14748739 TI - What does the lipoprotein oxidation phenomenon mean? AB - The evidence that oxidative lipid modification may be involved in the genesis of common diseases, such as atherosclerosis, is persuasive, but it was, until recently, conjecture based on in vitro findings, or investigation using experimental animal models. Recent clinical intervention studies in patients at high risk of cardiovascular events have been, at best, inconclusive. This has led to a general consensus that antioxidant supplements are of no value in the prevention of cardiovascular disease in subjects at high risk. However, epidemiological studies have demonstrated that the protective effects of antioxidant supplements, specifically vitamin E, were particularly evident amongst healthy subjects taking supplements. The picture is further clouded by the uncertain mechanism of lipoprotein modification within the artery wall, the possibility that some antioxidants may, under certain conditions, become pro oxidants, the complex interactions between lipid- and water-soluble antioxidants, and the fact that free-radical-mediated events may only be important in the early stages of atherogenesis. Recent results also suggest that the biological efficacy of antioxidants, such as alpha-tocopherol, may be compromised by the conditions extant within the plaque. It is evidently important that the position on the benefits of antioxidants, whether in food or as supplements, in disease prevention is clarified. PMID- 14748740 TI - Glycosylation of the receptor guanylate cyclase C: role in ligand binding and catalytic activity. AB - GC-C (guanylate cyclase C) is the receptor for heat-stable enterotoxins, guanylin and uroguanylin peptides. Ligand binding to the extracellular domain of GC-C activates the guanylate cyclase domain leading to accumulation of cGMP. GC-C is expressed as differentially glycosylated forms in HEK-293 cells (human embryonic kidney-293 cells). In the present study, we show that the 145 kDa form of GC-C contains sialic acid and galactose residues and is present on the PM (plasma membrane) of cells, whereas the 130 kDa form is a high mannose form that is resident in the endoplasmic reticulum and serves as the precursor for the PM associated form. Ligand-binding affinities of the differentially glycosylated forms are similar, indicating that glycosylation of GC-C does not play a role in direct ligand interaction. However, ligand-stimulated guanylate cyclase activity was observed only for the fully mature form of the receptor present on the PM, suggesting that glycosylation had a role to play in imparting a conformation to the receptor that allows ligand stimulation. Treatment of cells at 20 degrees C led to intracellular accumulation of a mature glycosylated form of GC-C that now showed ligand-stimulated guanylate cyclase activity, indicating that localization of GC-C was not critical for its catalytic activity. To determine if complex glycosylation was required for ligand-stimulated activation of GC-C, the receptor was expressed in HEK-293 cells that were deficient in N acetylglucosaminyltransferase 1. This minimally glycosylated form of the receptor was expressed on the cell surface and could bind a ligand with an affinity comparable with the 145 kDa form of the receptor. However, this form of the receptor was poorly activated by the ligand. Therefore our studies indicate a novel role for glycosidic modification of GC-C during its biosynthesis, in imparting subtle conformational changes in the receptor that allow for ligand mediated activation and perhaps regulation of basal activity. PMID- 14748741 TI - An inactive protein phosphatase 2A population is associated with methylesterase and can be re-activated by the phosphotyrosyl phosphatase activator. AB - We have described recently the purification and cloning of PP2A (protein phosphatase 2A) leucine carboxylmethyltransferase. We studied the purification of a PP2A-specific methylesterase that co-purifies with PP2A and found that it is tightly associated with an inactive dimeric or trimeric form of PP2A. These inactive enzyme forms could be reactivated as Ser/Thr phosphatase by PTPA (phosphotyrosyl phosphatase activator of PP2A). PTPA was described previously by our group as a protein that stimulates the in vitro phosphotyrosyl phosphatase activity of PP2A; however, PP2A-specific methyltransferase could not bring about the activation. The PTPA activation could be distinguished from the Mn2+ stimulation observed with some inactive forms of PP2A, also found associated with PME-1 (phosphatase methylesterase 1). We discuss a potential new function for PME 1 as an enzyme that stabilizes an inactivated pool of PP2A. PMID- 14748742 TI - Bcl2-low-expressing MCF7 cells undergo necrosis rather than apoptosis upon staurosporine treatment. AB - We present a ribozyme-based strategy for studying the effects of Bcl2 down regulation. The anti-bcl2 hammerhead ribozyme Rz-bcl2 was stably transfected into MCF7 cancer cells and the cleavage of Bcl2 mRNA was demonstrated using a new assay for cleavage product detection, while Western blot analysis showed a concomitant depletion of Bcl2 protein. Rz-bcl2-expressing cells were more sensitive to staurosporine than control cells. Moreover, both molecular and cellular read-outs indicated that staurosporine-induced cell death was necrosis rather than apoptosis in these cells. The study of the effects of Bcl2 down regulation was extended to the global MCF7 protein expression profile, exploiting a proteomic approach. Two reference electro-pherograms of Rz-bcl2-transfected cells, one with the ribozyme in a catalytically active form and the other with the ribozyme in a catalytically inactive form, were obtained. When comparing the two-dimensional maps, 53 differentially expressed spots were found, four of which were identified by MALDI-TOF (matrix-assisted laser-desorption ionization-time-of flight) MS as calreticulin, nucleophosmin, phosphoglycerate kinase and pyruvate kinase. How the up-regulation of these proteins might help to explain the modification of Bcl2 activity is discussed. PMID- 14748743 TI - STAT-1: a novel regulator of apoptosis. AB - Extracellular signalling molecules binding to their specific receptors are able to modulate gene expression, leading to changes in development, cell growth and homeostasis. The signal transducers and activators of transcription (STAT) protein family members are among the best studied of the latent cytoplasmic signal-dependent transcription factors. The STAT factors are activated via phosphorylation on the C-terminal domain following cytokine signalling or by various stress-induced stimuli. Recently, STAT-1 has been implicated in modulating pro- and anti-apoptotic genes following several stress-induced responses. These effects are dependent on STAT-1 phosphorylation on serine-727 and require the C-terminal transactivation domain of STAT-1 to enhance its pro apoptotic effect or inhibit its anti-apoptotic effects. The STAT-1 C-terminal domain has been demonstrated to be important for protein-protein interaction with other transcriptional activators. The reports that STAT-1-deficient mice develop spontaneous and chemically induced tumours more rapidly compared to wild-type mice and that STAT-1-deficient cells are more resistant to agents that induce apoptosis strongly support the argument that STAT-1 acts as a tumour suppressor. PMID- 14748744 TI - Production of a new model of slowly progressive Heymann nephritis. AB - A slowly progressive autoimmune kidney disease was induced in Sprague Dawley rats by subcutaneous injection of a chemically modified kidney antigen (rKF3), incorporated into Alum and Distemper complex vaccine, followed by subcutaneous injections of an aqueous preparation of the same antigen. Pathogenic autoantibodies developed, which reacted with fixed glomerular nephritogenic antigen. Subsequently, immunopathological events lead to chronic progressive immune complex glomerulonephritis and proteinuria. The slowly developing disease was morphologically and functionally similar to Heymann nephritis (HN). The damage observed in the kidneys of experimental animals at 8 weeks and at the end of the experiment was examined by direct fluorescent antibody test, histology and electron microscopy. The changes were similar to the typical lesions found in HN rat kidneys, but less severe. Animals became proteinuric from 17 weeks onward (instead of the usual 4-8 weeks). By the end of the experiment, at 8 months, 100% of the rats were proteinuric. This new experimental model of autoimmune kidney disease, which is not complicated by intraperitoneal deposition and retention of Freund's complete adjuvant and renal tubular antigens, allowed us to investigate the pathogenesis of the disease processes from a different aspect, and promises to be a useful and improved model for the investigation of future treatment options. PMID- 14748745 TI - Correlation of c-erbB-2 and S-100 expression with the malignancy grading and anatomical site in oral squamous cell carcinoma. AB - An immunohistochemical analysis of 32 cases of oral squamous cell carcinoma (eight in the inferior lip, eight in the lateral angle of the tongue, eight in the palate and eight in the mouth floor) was performed to evaluate the expression pattern of c-erbB-2 protein and S-100-positive cells in the lesions. The immunohistochemical expression was correlated with the tumour anatomical site and histological grading of malignancy. A higher frequency of c-erbB-2-positive cases was found in the tongue, even though no correlation could be detected between the protein expression and the tumour histological grading. With respect to the S-100 positive cells, it was observed that a quantitative decrease was present in the cases classified as high-grade tumours when compared to the low ones (P = 0.0007). Thus, c-erbB-2 immunohistochemical expression is correlated with anatomical localization, and the expression of the S-100 Langerhans' cell markers is decreased significantly in high-grade carcinomas. PMID- 14748746 TI - Targeted cellular process profiling approach for uterine leiomyoma using cDNA microarray, proteomics and gene ontology analysis. AB - This study utilized both cDNA microarray and two-dimensional protein gel electrophoresis technology to investigate the multiple interactions of genes and proteins involved in uterine leiomyoma pathophysiology. Also, the gene ontology analysis was used to systematically characterize the global expression profiles at cellular process levels. We profiled differentially expressed transcriptome and proteome in six-paired leiomyoma and normal myometrium. Screening up to 17 000 genes identified 21 upregulated and 50 downregulated genes. The gene expression profiles were classified into mutually dependent 420 functional sets, resulting in 611 cellular processes according to the gene ontology. Also, protein analysis using two-dimensional gel electrophoresis identified 33 proteins (17 upregulated and 16 downregulated) of more than 500 total spots, which was classified into 302 cellular processes. Of these functional profilings, downregulations of transcriptomes and proteoms were shown in cell adhesion, cell motility, organogenesis, enzyme regulator, structural molecule activity and response to external stimulus functional activities that are supposed to play important roles in pathophysiology. In contrast, the upregulation was only shown in nucleic acid-binding activity. Taken together, potentially significant pathogenetic cellular processes were identified and showed that the downregulated functional profiling has a significant impact on the discovery of pathogenic pathway in leiomyoma. Also, the gene ontology analysis can overcome the complexity of expression profiles of cDNA microarray and two-dimensional protein analysis via its cellular process-level approach. Therefore, a valuable prognostic candidate gene with relevance to disease-specific pathogenesis can be found at cellular process levels. PMID- 14748747 TI - High levels of intracellular ATP prevent nitric oxide-induced apoptosis in rat gastric mucosal cells. AB - Nitric oxide (NO) plays an important role in gastric mucosal injury in the human stomach. Exposure to excessive NO leads to apoptosis; however, the mechanism remains largely unknown in gastric epithelial cells. The apoptotic process is modulated by energy states in cells. This study investigated molecular mechanisms of NO-induced apoptosis in gastric epithelial cells and influence of high glucose on those mechanisms. Normal rat gastric mucosal epithelial (RGM-1) cells were cultured in media containing either 1000 (low) or 4500 mg/l (high) of D-glucose. When the cells were incubated with a chemical NO donor NOC18, apoptosis was induced in a dose-dependent manner. Intracellular adenosine triphosphate (ATP) levels significantly increased in the cells cultured with high glucose in comparison with the low-glucose condition. The cells with high ATP levels were more resistant to NO-induced apoptosis than the cells with low ATP levels. NO induced apoptosis was followed by mitochondrial depolarization, upregulation of Bax protein, cytochrome C release from mitochondria to the cytosol and subsequent caspases activation. These results suggest that NO inhibition of mitochondrial respiratory system and acute ATP depletion initiate apoptotic signalling in gastric epithelial cells. High glucose may prevent NO-induced apoptosis by leading to high levels of intracellular ATP or other metabolic changes in this cell line. PMID- 14748749 TI - Invited review: the evolution of antidepressant mechanisms. AB - Present antidepressants are all descendents of the serendipitous findings in the 1950s that the monoamine oxidase inhibitor iproniazid and the tricyclic antidepressant imipramine were effective antidepressants. The identification of their mechanism of action, and those of reserpine and amphetamine, in the 1960s, led to the monoamine theories of depression being postulated; first, with noradrenaline then 5-hydroxytryptamine being considered the more important amine. These monoamine theories of depression predominated both industrial and academic research for four decades. Recently, in attempts to design new drugs with faster onsets of action and more universal therapeutic action, downstream alterations common to current antidepressants are being examined as potential antidepressants. Additionally, the use of animal models has identified a number of novel targets some of which have been subjected to clinical trials in humans. However, monoamine antidepressants remain the best current medications and it may be some time before they are dislodged as the market leaders. PMID- 14748748 TI - Immunophenotyping of macrophages in human pulmonary tuberculosis and sarcoidosis. AB - Classic studies of tuberculosis (TB) revealed morphologic evidence of considerable heterogeneity of macrophages (MOs), but the functional significance of this heterogeneity remains unknown. We have used newly available specific antibodies for selected membrane and secretory molecules to examine the phenotype of MOs in situ in a range of South African patients with TB, compared with sarcoidosis. Patients were human immunodeficiency virus-negative adults and children, and the examined biopsy specimens included lung and lymph nodes. Mature pulmonary MOs (alveolar, interstitial, epithelioid and multinucleated giant cells) selectively expressed scavenger receptor type A and a novel carboxypeptidase-like antigen called carboxypeptidase-related vitellogenin-like MO molecule (CPVL). CPVL did not display enhanced expression in sarcoidosis, vs. TB patients, as observed with angiotensin-converting enzyme (ACE), a related molecule. Immunocytochemical studies with surfactant proteins (SP)-A and -D showed that type II alveolar cells expressed these collectins, as did MOs, possibly after binding of secreted proteins. Studies with an antibody specific for the C-terminus of fractalkine, a tethered CX3C chemokine, confirmed synthesis of this molecule by bronchiolar epithelial cells and occasional endothelial cells. These studies provide new marker antigens and extend previous studies on MO differentiation, activation and local interactions in chronic human granulomatous inflammation in the lung. PMID- 14748750 TI - Trends in peptic ulcer pharmacotherapy. AB - Considering the diseases of the stomach and duodenum, peptic ulcer has been the one with a significant clinical impact. The pathophysiology of peptic ulcer has centred on an imbalance between aggressive and protective factors. The discovery of Helicobacter pylori as a cause of peptic ulcer has changed our approach greatly towards this disease. Despite the decreasing frequency of H. pylori induced peptic ulcers, peptic ulcer remains a major clinical problem partly because nonsteroidal anti-inflammatory drug (NSAID)-related ulcers and hospital admissions for ulcer complications associated with NSAIDs have increased in frequency. The interaction between H. pylori and NSAIDs is one of the most controversial issues in peptic ulcer. In this article, current concepts of peptic ulcer etiopathogenesis and the management of peptic ulcer according to the etiology were reviewed. PMID- 14748751 TI - Synergism of atenolol and amlodipine on lowering and stabilizing blood pressure in spontaneously hypertensive rats. AB - This study was designed to investigate the possible synergism of atenolol and amlodipine on lowering and stabilizing blood pressure (BP) in spontaneously hypertensive rats. Sixty-four spontaneously hypertensive rats were randomly divided into eight groups. They were given 0.8% carboxymethylcellulose sodium (control), atenolol (10 mg/kg), amlodipine (0.5, 1 and 2 mg/kg) and the combinations of atenolol and amlodipine (10 + 0.5, 10 + 1 and 10 + 2 mg/kg), respectively. The drugs were given via a catheter of gastric fistula. BP was recorded for 25 h from 1 h before drug administration to 24 h after administration, in conscious, freely moving rats. It was found that combination of atenolol and amlodipine significantly decreased BP and systolic BP variability. From probability sum analysis it was found that the combination of atenolol and amlodipine, in a proportion as 10 : 1, was the best one (q = 1.54). In conclusion, the present work clearly demonstrated that there is a synergistic effect between atenolol and amlodipine in lowering and stabilizing the BP. The synergistic effect is highest when the dose proportion of the two drugs is 10 : 1. PMID- 14748752 TI - Myocardial hypertrophy is not a prerequisite for changes in early gene expression in left ventricular volume overload. AB - Currently it is not certain whether hypertrophy or the underlying disease is the primary trigger of the alterations in early gene expression in the progression of cardiac disease to end-stage heart failure. In this study, we tested the notion that in left ventricular overload disorders, the changes in early gene expression in the progression to heart failure is independent of the manifestation of cardiac hypertrophy. We compared the expression of the early genes c-fos, c-myc, and c-jun in six dilated cardiomyopathic hearts (DCM) and 15 patients with left ventricular volume overload (VOL) resulting from mitral/aortic regurgitation and no significant stenosis or hypertrophic manifestations, using eight healthy donor hearts as controls. In VOL, c-myc was elevated by 88% (P < 0.01) in the left ventricle, 46% in the right ventricle, onefold (P < 0.01) in the left atrium, and 54% (P < 0.05) in the right atrium, while in DCM, it was increased by 71% (P < 0.02), 55%, 48% (P < 0.05) and 91% (P < 0.05), respectively. Similarly, c-jun was elevated by 41% (P < 0.01) in the left ventricle, 39% (P < 0.05) in the right ventricle, 83% (P < 0.02) in the left atrium and 21% in the right atrium in VOL, while in DCM it was elevated by 13% in the left ventricle, 29% in the left atrium, and 41% in the right atrium, but decreased by 13% in the right ventricle. In contrast, c-fos was slightly decreased in the left ventricle and atrium of both DCM and VOL, and in left atrium of the VOL group, but remained unchanged in the other myocardial chambers. These results show that, in the human myocardium, the three early genes are regulated differently, possibly in disease- and chamber specific fashions, and manifestation of left ventricular hypertrophy is not a prerequisite for the elevation in their expression in left ventricular overload disorders. PMID- 14748753 TI - Vasopressin-induced vasoconstriction is dependent on MAPKerk1/2 phosphorylation. AB - To investigate the involvement of the mitogen-activated protein kinase (MAPK) family of extracellular signal-regulated kinase (ERK) 1 and 2 (MAPKerk1/2) in the vasopressin-mediated vasoconstriction in the rat aorta. Vasopressin-induced vasoconstriction was measured in isolated rat thoracic aortae in the presence or absence of MAPKerk1/2 kinase (MKKmek1/2) inhibitors. Thereafter the MAPKerk1/2 phosphorylation in the rat aorta was quantified using Western blot analysis. Vasopressin (1-300 nm) induced a concentration-dependent vasoconstriction, which could be inhibited concentration dependently by the selective MKKmek1/2 inhibitors, PD 98059 (10 and 100 microm) and U 0126 (10 and 100 microm). Western blot analysis revealed a 2.7 +/- 0.6-fold increase in the MAPKerk1/2 phosphorylation induced by vasopressin (300 nm). This phosphorylation could be dose dependently prevented by both PD 98059 (100 microm) and U 0126 (10 and 100 microm). These results indicate that vasoconstriction induced by vasopressin is partly regulated by the MAPKerk1/2 pathway. PMID- 14748754 TI - Lamotrigine kidney distribution in male rats following a single intraperitoneal dose. AB - As it has been previously shown that lamotrigine (LTG) accumulates in the kidney of male rats, the purpose of the present investigation was to characterize the kidney profiles of LTG and its kidney distribution pattern in male rats, in order to confirm if a preferential distribution exists and to analyse if it does or does not affect the LTG systemic pharmacokinetics. Adult male Wistar rats were intraperitoneally injected with 5, 10 and 20 mg/kg of LTG. The concentration-time profiles of LTG in plasma and whole kidney were determined over 120 h postdose. The distribution of LTG in the rat kidney was investigated in another group of rats by measuring LTG levels in the renal cortex and medulla. The LTG plasma concentration-time profiles revealed a linear relationship with dose. However, a slight increase in the LTG elimination half-life with dose was observed. In contrast, a nonlinear relationship was established between LTG kidney levels and the dose administered. Consequently, nonparallel patterns were observed between LTG plasma and kidney profiles. The LTG kidney distribution pattern revealed an accumulation of LTG in the renal cortex. The present study demonstrated that LTG distributes preferentially to the kidneys of the male rat in a dose-dependent manner and suggests that such distribution may slightly affect the systemic kinetics of the drug. PMID- 14748755 TI - Influence of simulated weightlessness on the pharmacokinetics of acetaminophen administered by the oral route: a study in the rat. AB - During space flights, the human body is submitted to weightlessness which induces physiological variations that could modify drug disposition during space missions. Since space experiments are infrequent and difficult to perform, in order to evaluate pharmacokinetic modifications, simulation experiments of weightlessness have to be carried out on earth, using animal-models such as the Morey-Holton model. In this model, rats are suspended by the tail with their front paws on the ground. We studied the effects of simulated weightlessness on drug absorption and on gastric emptying, using acetaminophen as a probe. Three periods of suspension (1, 2 and 5 days) were compared with two control groups (free and attached rats). The attached group was used to evaluate a possible 'stress effect' caused by the suspension device. Each group was composed of 36 rats (12 sampling times and three rats per time). An oral dose of acetaminophen (100 mg/kg) was administered and blood samples were collected before and up to 12 h after administration. Plasma assays were performed using an high-performance liquid chromatography method with UV detection. The calculated population pharmacokinetic parameters were Ka, Kel (first order absorption and elimination constants) and Vd/F (apparent volume of distribution). The statistical interpretation of the population pharmacokinetic parameters indicated that 2 days of suspension significantly decreased the Vd/F by 83% and the Ka by 125%. The increase in the Ka was probably because of an increased acceleration of the gastric emptying and/or to a decrease in the total peripheral resistance which increased intestinal blood flow. PMID- 14748756 TI - Diffusion of arylpropionate non-steroidal anti-inflammatory drugs into the cerebrospinal fluid: a quantitative structure-activity relationship approach. AB - A quantitative structure-activity relationship (QSAR) analysis of a series of arylpropionic acid non-steroidal anti-inflammatory drugs (NSAIDs) has been performed to determine which physicochemical properties of these compounds are involved in their diffusion into the cerebrospinal fluid (CSF). The penetration of eight arylpropionic acid derivatives into CSF was studied in male Wistar rats. After intraperitoneal administration of each compound (5 mg/kg), blood and CSF samples were collected at different times (0.5, 1, 3 and 6 h). The fraction unbound to plasma protein was determined using ultrafiltration. The areas under the curve of the free plasma (AUCF) and CSF (AUCCSF) concentrations were calculated according to the trapezoidal rule. The overall drug transit into CSF was estimated by the ratio RAUC (AUCCSF : AUCF). The lipophilicity was expressed as the chromatographic capacity factor (log kIAM) determined by high-performance liquid chromatography on an immobilized artificial membrane (IAM) column. A significant parabolic relationship was sought between lipophilicity (log kIAM) and the capacity of diffusion across the blood-brain barrier (log RAUC) (r = 0.928; P < 0.01). The arylpropionic acid NSAIDs exhibiting a lipophilicity value between 1.1 and 1.7 entered the CSF easily (RAUC > 1). The molecular weight (MW) was included in this parabolic relationship by means of a multiple regression analysis. This physicochemical parameter improved the correlation (r = 0.976; P < 0.005). Based on our findings, diffusion of arylpropionic acid NSAIDs into CSF appears to depend primarily on their lipophilicity and MW. PMID- 14748757 TI - Centrally injected CDP-choline increases plasma vasopressin levels by central cholinergic activation. AB - In the present study, both the effects of intracerebroventricular (i.c.v.) injection of cytidine-5'-diphosphate choline (CDP-choline) on plasma vasopressin levels and the choline involvement of these effects were investigated. I.c.v. administration of CDP-choline (0.5, 1.0 and 2.0 micromol) increased plasma vasopressin levels dose- and time-dependently. I.c.v. injection of equimolar dose of choline (1 micromol) produced similar vasopressin response. However equimolar dose of cytidine (1 micromol; i.c.v.), the other hydrolysis product of CDP choline, did not affect plasma vasopressin levels. Pretreatment of rats with hemicholinium-3, neuronal high affinity choline uptake inhibitor (20 microg; i.c.v.) blocked the vasopressin response to i.c.v. CDP-choline (1 micromol). Pretreatment of rats with mecamylamine (50 microg; i.c.v.), a nonselective nicotinic receptor antagonist, abolished the increase in plasma vasopressin induced by CDP-choline while atropine (10 microg; i.c.v.), nonselective muscarinic receptor antagonist, failed to change the response. In conclusion, intracerebroventricularly injected CDP-choline can increase plasma vasopressin levels by activating central nicotinic cholinergic receptors through the activation of presynaptic cholinergic mechanisms. PMID- 14748758 TI - Effect of tempol (4-hydroxy tempo) on gentamicin-induced nephrotoxicity in rats. AB - We investigated the effects of tempol (4-hydroxy tempo), a membrane-permeable radical scavenger, on gentamicin-induced renal failure in rats. The rats were given gentamicin (100 mg/kg/day, i.p., once a day); and gentamicin (100 mg/kg/day, i.p.) and tempol (3.5, 7 or 14 mg/kg/day, i.p., once a day). At the end of 7 days, the gentamicin group produced the remarkable nephrotoxicity, characterized by a significantly decreased creatinine clearance and increased serum creatinine, blood urea nitrogen (BUN) and daily urine volume when compared with controls. In control the BUN value was 21.2 +/- 0.07 (mg/100 mL); in comparison, it was 96.9 +/- 6.03 in gentamicin group (P < 0.05). Renal histopathologic examination confirmed acute tubular necrosis in this group. In rats treated with gentamicin and tempol a partial improvement in biochemical and histologic parameters was observed. BUN values were 96.9 +/- 6.03 and 36.3 +/- 2.39 in gentamicin, and gentamicin plus tempol (14 mg/kg) treated groups, respectively (P < 0.05). These results suggest that the administration of tempol may have a protective effect on gentamicin-induced nephrotoxicity in rats. PMID- 14748759 TI - Gemcitabine-induced severe pulmonary toxicity. AB - Gemcitabine is a relatively new deoxycytidine analog (2',2' difluorodeoxycytidine) with structural similarities to cytosine arabinoside (Ara C). Activity of gemcitabine is demonstrated in the treatment of many solid tumors, like pancreas, ovarian and nonsmall cell lung cancer (NSCLC). Although gemcitabine is considered as a drug with a good safety profile, cases of gemcitabine-induced severe pulmonary toxicity (GISPT) were reported as for Ara-C. We performed a systematic review of reported cases on the GISPT. Twenty-nine clinical trials especially interesting NSCLC patients (21) and 21 reported cases recording 40 patients were analyzed. The incidence of the GISPT varies from 0 to 5%. The clinical presentation is a subacute clinical syndrome and is frequently nonspecific. The predominant radiographic pattern on chest X-ray are reticulo nodular interstitial infiltrates. It was postulated that the physio-pathological mechanism of the GISPT was an inflammatory reaction of the alveolar capillary wall cytokine-mediated, which created an abnormal permeability of its membrane. After the differential diagnosis were ruled out, the discontinuation of the drug and the early initiation of steroids and diuretics are the most frequently performed treatments. Under these conditions, the outcome was favorable in a delay of few days generally for a majority of patients but 20% of patients died. Some risk factors, as a previous pulmonary disease or a previous thoracic irradiation, for the occurrence of the GISPT were proposed. GISPT is rare but sometimes fatal. Its a necessity to increase awareness about it to enhanced an early and suitable management of patients developing such a toxicity after gemcitabine administration. PMID- 14748760 TI - Plasma antioxidative activity during atorvastatin and fluvastatin therapy used in coronary heart disease primary prevention. AB - We estimated the effect of atorvastatin and fluvastatin on plasma antioxidative activity used in coronary heart disease (CHD) primary prevention. Anti-oxidative activity of blood plasma was determined by Bartosz et al. method [Curr. Top. Biophys. (1998)22:11-13], based on reduction of preformed cation radical of 2,2,azinobis(3-ethylbenzothiazoline-6-sulphonic acid) by blood plasma. The study comprised 35 patients with CHD risk who were randomly divided into two groups. The atorvastatin group comprised 17 patients who were administered the drug orally in a daily dose of 10 mg and the fluvastatin group consisted of 18 patients on an oral dose of 40 mg once daily. The control group comprised 12 healthy subjects with no drug administration. Blood samples were collected from cubital vein before and after 6-week therapy. Significantly (P < 0.05) increased- in comparison with the initial values--antioxidative activity of blood plasma was found in atorvastatin and fluvastatin groups after 6-week therapy. Moreover, the increase in antioxidative plasma activity in atorvastatin group was significantly higher in comparison with the fluvastatin group. The results of our study have demonstrated that atorvastatin and fluvastatin have an additional mechanism independent of the effect on cholesterol concentration. Thus, we presume that administration of these statins in CHD risk patients may have a beneficial effect. PMID- 14748761 TI - Vinorelbine-related cardiac events: a meta-analysis of randomized clinical trials. AB - Several cases of cardiac adverse reactions related to vinorelbine (VNR) have been reported in the literature. In order to quantify the incidence of these cardiac events, we performed a meta-analysis of clinical trials comparing VNR with other chemotherapeutic agents in the treatment of various malignancies. Randomized clinical trials comparing VNR with other drugs in the treatment of cancer were searched in Medline, Embase, Evidence-based Medicine Reviews databases and the Cochrane library from 1987 to 2002. Outcomes of interest were severe cardiac events, toxic deaths and cardiac event-related deaths reported in each publication. We found 19 trials, involving 2441 patients treated by VNR and 2050 control patients. The incidence of cardiac events with VNR was 1.19% [95% confidence interval (CI) (0.75; 1.67)]. There was no difference in the risk of cardiac events between VNR and other drugs [odds ratio: 0.92, 95% CI (0.54; 1.55)]. The risk of VNR cardiac events was similar to vindesine (VDS) and other cardiotoxic drugs [fluorouracil, anthracyclines, gemcitabine (GEM) em leader ]. Even if it did not reach statistical significance because of a few number of cases, the risk was lower in trials excluding patients with cardiac history, and seemed to be higher in trials including patients with pre-existing cardiac diseases. Vinorelbine-related cardiac events concern about 1% of treated patients in clinical trials. However, the risk associated with VNR seems to be similar to that of other chemotherapeutic agents in the same indications. PMID- 14748762 TI - Undocumented drug utilization and drug waste increase costs of pediatric anesthesia care. AB - The present study was performed in order to identify the cost of drugs used without documenting them in the patients' file and the wastage of drugs in a pediatric anesthesiology ward. In a prospective, blinded, observational design, drug utilization of 610 consecutive patients, undergoing an elective or emergency surgical procedure was evaluated. The number of undocumented drugs per 100 requested units and the number of wasted drugs per 100 requested units were computed and multiplied by its corresponding unitary cost. The median undocumented cost was 92.4 US dollars (95% CI 17.2-216.6 dollars) per 100 requested units. Succinylcholine (40 mg/2 mL) was the main undocumented drug; its use was not documented in approximately 50% cases in which this neuromuscular blocking agent was requested. However, rocuronium and nalbuphine had the highest unjustified cost, 770.6 dollars and 847.0 dollars per 100 requested units, respectively. Ketorolac, diclofenac, metamizol, furosemide, methylprednisolone, sodium bicarbonate, and cisatracurium were requested and documented. The median cost of wasted drug was 141.8 dollars (95% CI 55.8-448.2 dollars) per 100 requested drugs. More than 80% of adrenaline, naloxone, flunitrazepam, ephedrine, and cisatracurium were wasted. However, the highest cost of wasted drugs was for ondansetron, cisatracurium, methylprednisolone, and rocuronium. The uncontrolled availability and use of drugs may represent an important amount of resources wasted without any awareness of the staff in a department of pediatric anesthesia. PMID- 14748763 TI - Pharmacokinetic and pharmacodynamic interactions between metoprolol and dronedarone in extensive and poor CYP2D6 metabolizers healthy subjects. AB - As dronedarone a new noniodinated antiarrhythmic agent structurally related to amiodarone could inhibit CYP2D6 and is planned to be associated with beta blockers, interactions with CYP2D6 metabolized beta-blockers such as metoprolol, have to be studied. Forty-nine healthy male subjects genotyped for CYP2D6 were included in a randomized, double-blind, placebo-controlled study. Metoprolol was administrated during 13 days (200 mg/day). After the initial 5 days, subjects received placebo (n = 12), 800 mg (n = 6), 1200 mg (n = 9), or 1600 mg (n = 17) of dronedarone daily during eight additional days. Pharmacokinetic parameters of metoprolol were investigated at day 5 and at day 13 in 44 subjects, 39 extensive metabolizers and five poor metabolizers for CYP2D6. Cardiac contractility function was evaluated by the rate-corrected electromechanical systole duration (QS2i) and the mean velocity of endocardial circumferential fiber shortening (Vcfmean). Cmax and AUC0--24 h of metoprolol increased from days 5 to 13 in proportion to dronedarone dose only in CYP2D6 extensive metabolizers genotyped subjects (P < 0.001). In all subjects, from days 5 to 13, Vcfmean decreased and QS2i significantly increased in dronedarone groups. The Vcfmean changes were however significant only with the 1600 mg dronedarone dose compared with placebo while QS2i changes induced by addition of dronedarone were significant compared with placebo at all dose levels. Between days 5 and 13, QS2i and Vcfmean changes were significantly correlated with both dronedarone concentrations at day 13 and with metoprolol concentration changes between days 5 and 13. Plasma metoprolol concentrations were highest in poor metabolizer subjects and dronedarone did not further increase their level but increased QS2i in the two subjects receiving the 1600 mg dose. Addition of dronedarone (800-1600 mg daily) to metoprolol (200 mg daily) increases bioavailability of metoprolol in CYP2D6 extensive metabolizers and induces an additive dronedarone dose-dependent negative inotropic effect. Nevertheless at 800 mg daily (anticipated therapeutic dose) these effects were modest. PMID- 14748764 TI - Is licofelone, a dual inhibitor of cyclo-oxygenase and 5-lipoxygenase, a promising alternative in anti-inflammatory therapy? AB - As prostaglandins and leukotrienes are critical in inflammation, dual cyclo oxygenase and 5-lipoxygenase enzymes inhibitors, especially licofelone, are being developed by pharmaceutical companies. Experimental data indicate that licofelone shares the antipyretic, analgesic, anti-inflammatory and anti-platelet activities of conventional nonsteroidal anti-inflammatory drugs (NSAIDs), and exhibits anti allergic properties. Although licofelone may lead to similar adverse effects on the kidney than available NSAIDs, it appeared to induce less gastrointestinal damaging effects than nonselective NSAIDs in animals. Unfortunately, preliminary clinical studies provided less impressive data with respect to efficacy. Finally, the experimental promise of licofelone as a safe and potent anti-inflammatory and analgesic agent remains to be proved in humans. PMID- 14748765 TI - Rationale, design and methods of the CASHMERE study. AB - Carotid intima-media thickness (IMT) measurement is a noninvasive method used for quantification of early stage of atherosclerosis. Data suggest that the combination of statin and hormone replacement therapy (HRT) might be useful in reducing the early progression of atherosclerosis in postmenopausal women. The main aim of the study is to compare the effects of 12-month therapy with atorvastatin (80 mg/day), HRT (oral 17beta-estradiol 1 or 2 mg/day, plus cyclic dydrogesterone 10 mg) alone and their combination vs. placebo on the progression of carotid IMT by using a high-definition echotracking device. The secondary objectives are to assess the effects of the treatments vs. placebo on arterial stiffness, lipid profile and C-reactive protein. The CASHMERE trial is an European randomized study with a 2 x 2-factorial design, double blinded for atorvastatin and prospective randomized, open blinded endpoint evaluation (PROBE) method applied to HRT. The investigators can adjust the dose of estradiol at any time during follow-up if necessary. A total of 800 postmenopausal women with mild hypercholesterolemia and with no previous history of cardiovascular disease will be included and followed up by their physicians [general practitioners (GPs) or gynecologists] for 1 year. The CASHMERE trial is the first randomized clinical trial to examine the effects of a statin alone or combined with HRT on the structure and the function of carotid artery as early markers of atherosclerosis in postmenopausal women with mild hypercholesterolemia. The results are expected for 2007. PMID- 14748766 TI - Spinal cord injury pain--mechanisms and treatment. AB - In spinal cord injury (SCI), pain is a major cause of disability. A review of experimental and human studies, which provide insight into the mechanisms and treatment of SCI neuropathic pain are presented. Each of a series of pathophysiologic changes after SCI may be relevant for the development of SCI neuropathic pain. These changes are discussed in relation to neuropathic pain at and below the level of SCI. SCI neuropathic pain is difficult to treat. Experimental and human randomized, double-blind, placebo-controlled, clinical trials on pharmacologic treatment of SCI pain are summarized. PMID- 14748767 TI - EFNS task force--therapy of nystagmus and oscillopsia. AB - An overview of possible treatment options for oculomotor disorders that prevent clear vision is given. Downbeat nystagmus, upbeat nystagmus, seesaw nystagmus, periodic alternating nystagmus, acquired pendular nystagmus, and saccadic oscillations such as opsoclonus/ocular flutter are discussed. In addition, superior oblique myokymia and vestibular paroxysmia are reviewed. All treatment recommendations available in the literature are classified as class C only. In general, only some of the patients benefit from the treatment. PMID- 14748768 TI - Agreement between the clinical Oxfordshire Community Stroke Project classification and CT findings in Poland. AB - The Oxfordshire Community Stroke Project (OCSP) classification provides a simple means of classifying the clinical syndromes associated with acute stroke. The validity of the classification can depend on many factors. Accuracy and time of the clinical and radiological examination are very important. It was used in the International Stroke Trial (IST). The study was conducted in 467 hospitals in 34 countries. Our aim was to assess how well the OCSP classification could predict infarct site and size on computed tomography (CT) scan when performed in a trial within 48 h after the onset of stroke and the clinical assessment was carried out by different doctors in different hospitals. We examined data on the patients randomized in the IST by the seven participating hospitals in Poland. Patients admitted to the hospital were examined by the doctor on duty. Eight aspects of the neurological deficit present just before randomization were recorded. The computer system in the randomization centre employed a validated algorithm to assign the patient to one of the four infarct syndromes: lacunar syndrome (LACS), partial anterior circulation syndrome (PACS), total anterior circulation syndrome (TACS) and posterior circulation syndrome (POCS). We assessed the localization and extent of the recent infarction on available CT scans and correlated these with the computer-assigned OCSP category. CT scans were available for 558/759 (74%) of the patients randomized in Poland. In 458 (82%) of cases, CT was carried out in the first 24 h after the onset of stroke. In 444 (80%) scans, a recent infarct was visible. These radiological lesions were appropriate to the clinical classification in 56% of patients with TACS, 73% with PACS, 61% with LACS and 59% with POCS. In Polish centres in IST, amongst the patients with infarction visible on CT, the OCSP subtype predicted the size and site of the infarct in about two thirds of cases. These data suggest that, provided its limitations are taken into account, the classification can be usefully applied in multicentre clinical trials (or epidemiological studies) and to aspects of the routine clinical care of patients with acute stroke. PMID- 14748769 TI - Risk modifiers for peripheral sensory neuropathy in HIV infection/AIDS. AB - The objective of this study is to examine the risk factors associated with the development of sensory neuropathy in human immunodeficiency virus (HIV)-infected patients in 292 HIV+ patients recruited through a community-based sentinel survey. We determined the clinical and treatment factors associated with the presence of peripheral sensory neuropathy in HIV+ subjects at baseline examination, and at 1-year follow-up. Baseline examination was assessed with a logistic regression analysis controlling for age, education level, history of drug/alcohol use, and anti-retroviral treatment. The risk of developing new peripheral neuropathy at follow-up was determined using a Cox proportional hazard model analysis. At study entry, neuropathy (n=64) was associated with acquired immunodeficiency syndrome (AIDS), nucleoside analogue reverse transcriptase inhibitors (NRTI) (i.e. ddC), and history of alcohol abuse. After 1-year follow up, the development of neuropathy was predicted by AIDS, age (older subjects), and NRTI use. These findings indicated that AIDS, age, alcohol abuse/dependence, and anti-retroviral medication use are important predictors of motor/sensory peripheral neuropathy in the HIV infection. The peripheral neurotoxic effect of anti-retroviral medication should be taken into account in the design of long term therapies. PMID- 14748770 TI - Spasms in children with definite and probable mitochondrial disease. AB - The diagnosis of mitochondrial encephalomyopathies is complex and a system for classification of the diagnosis as definite, probable, and possible has been proposed. The objective of this study was to explore the spectrum of epileptic disorders associated with probable and definite mitochondrial disease in children using this classification system. The patient population with mitochondrial disease and epilepsy was selected from a tertiary care children's hospital. Interictal electroencephalograms and video-EEG recordings were used to characterize seizure types. Ten children fulfilled the criteria for probable or definite mitochondrial disease and had epilepsy. Four had siblings with a similar clinical phenotype. Spasms were the most common seizure type and were the initial seizure type in seven patients and two siblings. Four patients had only partial seizures, with or without generalization, and one patient had seizures that were difficult to classify. Blood lactate concentrations were elevated consistently in patients with partial seizures alone but were occasionally normal in children with spasms. Spasms were the most common presenting seizure type in children with probable and definite mitochondrial disease. PMID- 14748771 TI - An occipital lobe epileptogenic focus in a patient with West Nile encephalitis. AB - Although most human cases of West Nile (WN) fever are benign, approximately 1% produce severe neurological illness. Meningitis and/or encephalitis comprise 75% of hospitalized cases with seizures in 10-15%. Occipital lobe seizures, often mimicking other primary seizure types due to extra-occipital spread, is uncommon in adults and especially so from an infectious origin. A case of WN encephalitis presenting with a simple partial seizure, focal motor, resulting from an occipital epileptogenic focus is reported. The atypical epileptogenic location of the case and the observed frequency of seizures in WN encephalitis suggest that this virus is particularly irritative to cortical neuronal networks. Thus when seizures especially with atypical EEG patterns present during an acute febrile illness in the warmer months, WN encephalitis should be considered. PMID- 14748772 TI - Comparison of motor fluctuations and L-dopa-induced dyskinesias in patients with familial and sporadic Parkinson's disease. AB - The aim of the present work was to study the characteristics of motor complications in a group of familial Parkinson's disease (fPD) patients in comparison with matched sporadic PD (sPD) patients. Fifty-one fPD and 51 sPD patients matched for age and disease duration, used as controls, were included in the study. The Unified Parkinson's Disease Rating Scale was completed during clinical examination and all patients were questioned about the characteristics of motor complications. The mean time from start of L-dopa therapy to the onset of motor fluctuations (MF) and L-dopa-induced dyskinesias (LID) as well as the mean time from symptom onset to the development of MF and LID was significantly shorter in the fPD group of patients. An analysis revealed a higher occurrence of MF and LID in fPD patients in the group with disease duration of 5 years or less. FPD may be associated with a higher prevalence and earlier onset of motor complications during the initial stages of the disease. Genetic factors may contribute to the specific characteristics of motor complications in fPD. PMID- 14748773 TI - A clinical and genetic study of 56 Saudi Wilson disease patients: identification of Saudi-specific mutations. AB - Wilson disease (WD) is a hereditary disorder, with recessive transmission and genetic heterogeneity. Several mutations of ATP7B, the gene underlying WD, were reported in many ethnic groups. In this study, mutation screening in ATP7B of 56 Saudi Arabian WD patients was undertaken. The clinical data of all patients were recorded. The entire ATP7B coding sequence, including intron-exon boundaries were screened for mutation by the polymerase chain reaction (PCR)-based mutation detection technique and DNA sequencing. Thirty-nine patients were symptomatic at presentation and 17 subjects were pre-symptomatic siblings of affected patients. Fourteen patients had neurological, 11 patients had mixed (hepatic and neurological), and 14 patients had hepatic presentations. Family history suggestive of WD was present in 72% of cases and 68% had consanguineous parents. Genetic analysis showed disease-causing mutations in three exons (exons 8, 19 and 21) of the ATP7B gene in 28 patients (50%). Mutations in exons 21 (18 cases) and 19 (one case) were unique for Saudis. This large series of Saudi patients with WD has shown wide variability in the genomic substrate of WD. There is no correlation between genotype and clinical presentation. PMID- 14748774 TI - The 40-mg dose of eletriptan: comparative efficacy and tolerability versus sumatriptan 100 mg. AB - Meta-analysis provides valuable information regarding relative efficacies of triptans, but head-to-head comparator studies remain the gold standard. Three similar head-to-head trials comparing eletriptan 40 mg (E40) with sumatriptan 100 mg (S100) provide a rare opportunity and sufficient power, for robust comparisons of efficacy. Data were combined from three double-blind, placebo-controlled, first-dose, first-attack acute migraine treatment studies comparing E40 (n=1132), S100 (n=1129), and placebo (n=645). The primary outcome was headache response at 2 h. Secondary outcomes included headache response at 1 h, pain-free and functional responses, and sustained headache and pain-free responses. Odds ratios were calculated for summary estimates of probability of response. There were higher headache response rates with eletriptan versus sumatriptan at 2 h (67% vs. 57%; P<0.0001) and 1 h (34% vs. 26%; P<0.0001). Eletriptan also had higher 2 h pain-free (35% vs. 25%; P<0.0001) and functional responses (67% vs. 58%; P<0.0001). Sustained headache (42%) and pain-free (22%) response rates were higher for eletriptan versus sumatriptan (34%, P<0.0001; 15%, P<0.0001). The probability of response for eletriptan versus sumatriptan ranged from 36% higher (relief of nausea) to 64% higher (sustained pain-free rate). Combined analysis demonstrates that E40 has superior efficacy versus S100 across all clinically relevant outcomes. PMID- 14748775 TI - Adult post-infectious thalamic encephalitis: acute onset and benign course. AB - We report on two young patients with an encephalitic syndrome and bilateral thalamic lesions following a presumably viral or mycoplasma respiratory tract infection with the main clinical symptoms of organic psychosis in the first and a prolonged amnestic syndrome and ataxia in the second case. Four months later the patients had recovered clinically and the thalamic lesions had resolved on magnetic resonance imaging in one case and almost completely in the other. We interpret the patients' illness as rare cases of a post-infectious acute thalamic encephalitis in adults. The cases and their relationship to possible post infectious autoimmune inflammatory or toxic pathophysiological mechanisms are discussed and a review of the literature is provided. PMID- 14748777 TI - Familial hemiplegic migraine. PMID- 14748776 TI - The risk of cancer from azathioprine as a treatment for multiple sclerosis. PMID- 14748784 TI - Learning to swim with LBC. PMID- 14748785 TI - Monitoring and evaluating the performance of the UK NHS Cervical Screening Programme: monitoring performance by using cytology outcomes adjusted for population characteristics. AB - Current quality assurance measures used in the NHS cervical screening programme (NHSCSP) include a review of laboratories with percentages of moderate/severe and borderline/mild smear results outside the 10th-90th percentiles. The method is limited by the fact that many of these outlier smear percentages may reflect laboratories covering populations with low or high risk and/or short or long average screening intervals. This paper outlines a new approach to aid the detection of outlier laboratories, by using data collected at the primary care trust (PCT) or health authority (HA) level and making allowances for population characteristics and screening interval. The setting is the NHSCSP in England using annual data provided by HAs. Data from the screening year 2000-01 is used to illustrate the methodology, although the methods can also be applied to data at the PCT level (now being collected for 2002-03 onwards). Percentages of smear results have been analysed against a series of explanatory variables using logistic regression models. These explanatory variables include Townsend deprivation index, uptake-corrected ethnic minority composition, a measure of screening interval, area type and region. An expected percentage of borderline/mild and moderate/severe smears is estimated from the models and an observed : predicted ratio (OPRmod/sev and OPRbord/mild) calculated. Low values are suggestive of relative undercalling and high values overcalling, after allowance for population characteristics. Analysis of data for 2000-01 showed that the OPRmod/sev for the 99 HAs varied from 0.68 to 1.44. Laboratories with low percentages of moderate/severe smears, but associated with PCTs or HAs with OPRmod/sev values closer to unity may not need to be investigated as their observed rates are consistent with predicted rates based on population characteristics. The method could also be directly applied to laboratories if further information on the population covered by each laboratory were routinely collected. PMID- 14748786 TI - Randomized controlled trial evaluating rapid pre-screen of cervical cytology specimens. AB - A randomized controlled trial of 75,355 cervical cytology specimens was performed comparing rapid pre-screen with no pre-screen. While the percentage of cases receiving a final report of definite high-grade abnormality was higher in the no pre-screen arm (no pre-screen = 0.70%, pre-screen = 0.65%), the percentage of cases receiving a final report of possible or definite high-grade abnormality was essentially identical in the two arms of the trial (no pre-screen = 1.22%, pre screen = 1.21%). In the randomized trial, one extra cytology report of definite high-grade abnormality was made for every 12,568 slides pre-screened. This level of benefit was reduced by about half when rapid pre-screen was adopted as a routine laboratory practice. PMID- 14748787 TI - Clinicopathological significance of Borderline Nuclear Change - High Grade Dyskaryosis Not Excluded. AB - During a recent discussion on classification of cervical cytology, the introduction of a 'Borderline Nuclear Change - High Grade Dyskaryosis Not Excluded' (BNCH) category was proposed. BNCH cases diagnosed prospectively were retrieved from laboratory records. Questionnaires were sent to referring practitioners regarding clinicopathological outcome. Cytopathological features resulting in the BNCH classification were recorded on slide review. A total of 103 reports on conventional cervical smears diagnosed as BNCH from 1999 to 2002 were retrieved, comprising 0.096% of 107 634 smears. Of 86/103 cases with clinical follow-up, CIN2 or worse was present in 30 (35%); 15 (17%) showed a borderline/low-grade abnormality and 41 (48%) were negative. No individual cytopathological feature was predictive of high-grade disease on follow-up. The yield of high-grade abnormalities on follow-up of BNCH supports the introduction of this terminology. PMID- 14748789 TI - Role of fine needle aspiration cytology in the diagnosis of soft tissue tumours and tumour-like lesions. AB - In this study, we evaluated the usefulness of fine needle aspiration cytology (FNAC) in the diagnosis of soft tissue tumours. We have also assessed the various pitfalls of FNAC of soft tissue tumours. This was a retrospective study and here we analysed only 82 histopathology proven cases of FNAC of soft tissue tumours diagnosed in a five and half year period. On histopathological examination, 55 of these cases were malignant and 27 were benign. There was a total of 15 recurrences and histopathology was available prior to FNAC in only eight of these cases. Therefore, excluding these eight cases, malignant tumours were primarily diagnosed by FNAC in 47 cases. The sensitivity, specificity and positive predictive value of FNAC in diagnosis of soft tissue tumours were 91.5%, 92.5% and 95.5%, respectively. Only 22 of 47 cases (46.8%) were correctly categorized. There were two false-positive and four false-negative cases. One case each of fibromatosis and schwannoma were reported as sarcoma. False-negative cases were fibrosarcoma (1), malignant nerve sheath tumour (2) and haemangiopericytoma (1). FNAC was very useful in distinguishing benign from malignant soft tissue tumours. However, it was not so effective in exact categorization of tumours. PMID- 14748788 TI - Expression of p120, Ki-67 and PCNA as proliferation biomarkers in imprint smears of prostate carcinoma and their prognostic value. AB - The cell proliferation markers p120, Ki-67 and proliferating cell nuclear antigen (PCNA) recognize nuclear antigens. The expression of these proteins by immunostaining methods was reported to be of value in determining the prognosis of patients with malignant diseases. In this study, we evaluated the prognostic significance of the expression of nuclear antigens p120, PCNA and Ki-67 in prostate cancer and compared the results with other prognostic factors. Imprint smear samples obtained from 70 patients immediately after radical prostatectomy for prostatic carcinoma were immunostained with monoclonal antibodies against p120, Ki-67 and PCNA. The immunostaining results were correlated with Gleason score, tumour differentiation, stage and prostatic specific antigen (PSA) levels. Our findings demonstrate that p120, Ki-67 and PCNA expression in prostatic carcinoma smears, correlated significantly with the degree of Gleason score (P < 0.001). When combining p120, Ki-67 and PCNA positivity with tumour differentiation there was a significant association among these parameters (P < 0.001). Overexpression of p120, Ki-67 and PCNA, was also associated with increased PSA serum levels (>4 ng/ml) (P < 0.001). The distribution of p120, Ki 67 and PCNA expression in prostate carcinomas was not statistically significant for Ki-67 (P = 0.69) and p120 (P = 0.22) but was significant for PCNA (P < 0.001) as far as the histological stage (T2a, T2b, T2c, T3a). P120, Ki-67 and PCNA expression had significant prognostic value for disease-free survival. Our results conclude that nuclear antigens p120, Ki-67 and PCNA appear to be additional markers in the field of prognosis of prostatic carcinoma. PMID- 14748790 TI - Cytodiagnosis of neoplasms of the central nervous system in cerebrospinal fluid samples with an application of selective immunostains in differentiation. AB - In this study cytological findings in specimens of cerebrospinal fluid (CSF) of central nervous system (CNS) tumours (16 primaries, 57 metastatic and 12 suspicious) are presented, which were diagnosed over a period of 7 years in 85 patients (50 females and 35 males) with an age range of 2-76 years. The follow-up included information from clinicians and a review of medical charts, histological correlation and/or further investigations following cytodiagnosis. The patients clinically presented with signs and symptoms of meningeal involvement. The primary tumours included six medulloblastomas, eight gliomas (four glioblastomata multiforme, two anaplastic astrocytomas, and two ependymomas) and two germinomas. The metastatic tumours were 14 melanomas, 19 breast carcinomas, four leukaemias, six B-cell lymphomas, five adenocarcinomas of gastrointestinal origin, seven carcinomas of lung, one retinoblastoma and one neuroblastoma. Twelve cases were reported as suspicious. On further investigations, four of these were from a primary tumour (two glioblastomata multiforme and two anaplastic astrocytomas) while the other eight cases were of a metastasis (one B-cell lymphoma, three breast carcinomas, three melanomas and one adenocarcinoma of gastrointestinal origin). Using a panel of selective immunostains in some of the cases supported the cytological diagnosis and this was considered useful in furthering cytodiagnosis. In 75 of the patients the CSF samples were obtained on a spinal tap while in 10 patients the samples were received as ventricular CSF. There were no false-positive cases. The results of our study suggest that CSF cytology in the diagnosis of CNS tumours is quite reliable and reflects involvement of leptomeninges or the ventricles. Furthermore, the use of selective immunostains can be helpful in confirming the cytological impression and source of the tumour. PMID- 14748791 TI - Diagnosis of tuberculous lymphadenitis by FNAC, microbiological methods and PCR: a comparative study. AB - Despite its usefulness in the diagnosis of tuberculous lymphadenitis, fine needle aspiration cytology (FNAC) faces several limitations, and its sensitivity and specificity are not well established. The diagnostic accuracy and limitations of FNAC were studied in comparison with conventional microbiological methods and polymerase chain reaction (PCR). Sixty patients with lymphadenopathy and a clinical diagnosis of tuberculous lymphadenitis were subjected to FNA. The aspirate was used for cytological examination, Ziehl-Neelsen staining, mycobacterial culture and PCR. PCR was performed using two sets of oligonucleotide primers for Mycobacterium tuberculosis and a single primer for M. bovis species. The results of FNAC, microbiological methods and PCR correlated with the clinical outcome after follow-up for an average period of 24 months. Twenty-five cases (41.6%) were treated and responded well to anti-tuberculosis therapy, among them 17 were correctly diagnosed by FNAC (68%), eight by microbiological methods (32%) and 24 by PCR (96%). When PCR is considered the gold standard, FNAC predicted the correct diagnosis in 62% of cases with a high false negative rate (38%) due to the absence of granuloma/necrosis in smears from cases of early tuberculosis. In the latter group PCR proved to be the most valuable and a diagnostic success of 100% was achieved when FNAC and PCR were combined. In addition, PCR allowed immediate characterization of M. tuberculosis in the vast majority (96.2%) of cases in the study population. PMID- 14748792 TI - Ethical problems in cytology. AB - Great advances in medical science have raised a number of ethical issues, many of which affect cytopathology. Some of the main issues addressed in this paper relate to the organization of a cytology laboratory: internal and external quality control, adequate staffing levels and staff education, cytopathology reporting format and contents, confidentiality issues, relationship with the clinicians and involvement of cytopathologists in clinical management teams. Quality control has to be provided within cytology departments but external quality assurance is also essential, with national monitoring. New technologies should be used according to the best scientific methods, following cytological analysis. Scientific work in cytology has to respect the general principles of scientific ethics. The patient's interest has to be the main reason for such work. PMID- 14748793 TI - Cytopathology in Slovenia. AB - Cytopathology started in Slovenia in the early 50s with exfoliative cytology, while fine needle aspiration biopsy (FNAB) was introduced some 10 years later. Today cytopathology is a well accepted diagnostic method in Slovenia and there are currently 20 cytopathological laboratories and 17 cytopathologists. The number of specimens examined in 2001 was 26 230 FNABs, 13 355 exfoliative non cervical and 323 888 cervical smears. FNABs are performed by cytopathologists, by clinical doctors and by radiologists. So far only the cytopathologists have a supervised training period in performing biopsies. In future the same requirement will be obligatory for non pathologists. In four laboratories immunocytochemistry is used as an ancillary technique to morphology and one laboratory is using also flow cytometry for immunophenotyping of lymphomas. The classification system used in Slovenia for reporting the findings in cervical cytology is a combination of Papanicolaou's classification and assessment of dyskariosis. In spite of a long tradition in opportunistic screening for cervical cancer (Cca) an organised screening programme was started late. A four year pilot study, which included one third of the women population of Slovenia, was concluded in 2002 and an organised screening programme was introduced to the whole country in 2003. The incidence rate of Cca in Slovenia has been rising slowly since 1994 and it reached 19.6/100 000 in the year 2000. The mortality rate has remained roughly constant at 5-7/100 000 for the last 20 years. During the last few years quality assurance measures have been taken for improving the performance in cervical cytology. PMID- 14748794 TI - Educational case report - self-assessment. Three patients with unusual metastases. PMID- 14748795 TI - Abolish C3. PMID- 14748796 TI - Abolish C3. PMID- 14748798 TI - History of the development of azole derivatives. AB - Until the 1940s, relatively few agents were available for the treatment of systemic fungal infections. The development of the polyene antifungals represented a major advance in medical mycology. Although amphotericin B quickly became the mainstay of therapy for serious infections, its use was associated with infusion-related side-effects and dose-limiting nephrotoxicity. The continued search for new and less toxic antifungals led to the discovery of the azoles several decades later. Ketoconazole, the first available compound for the oral treatment of systemic fungal infections, was released in the early 1980s. For almost a decade, ketoconazole was regarded as the drug of choice in nonlife threatening endemic mycoses. The introduction of the first-generation triazoles represented a second major advance in the treatment of fungal infections. Both fluconazole and itraconazole displayed a broader spectrum of antifungal activity than the imidazoles and had a markedly improved safety profile compared with amphotericin B and ketoconazole. Despite widespread use, however, these agents became subject to a number of clinically important limitations related to their suboptimal spectrum of activity, the development of resistance, the induction of hazardous drug-drug interactions, their less than optimal pharmacokinetic profile (itraconazole capsules), and toxicity. In order to overcome these limitations, several analogues have been developed. These so-called 'second-generation' triazoles, including voriconazole, posaconazole and ravuconazole, have greater potency and possess increased activity against resistant and emerging pathogens, in particular against Aspergillus spp. If the toxicity profile of these agents is comparable to or better than that of the first-generation triazoles and drug interactions remain manageable, then these compounds represent a true expansion of our antifungal arsenal. PMID- 14748799 TI - Twelve years of fluconazole in clinical practice: global trends in species distribution and fluconazole susceptibility of bloodstream isolates of Candida. AB - We determined the species distribution and in-vitro susceptibility of 6082 bloodstream infection (BSI) isolates of Candida spp. collected from 250 medical centres in 32 nations over a 10-year period from 1992 through 2001. The species included 3401 C. albicans, 984 C. glabrata, 796 C. parapsilosis, 585 C. tropicalis, 153 C. krusei, 67 C. lusitaniae, 48 C. guilliermondii, 10 C. famata, 10 C. kefyr, six C. pelliculosa, five C. rugosa, four C. lipolytica, three C. dubliniensis, three C. inconspicua, two C. sake and one isolate each of C. lambica, C. norvegensis and C. zeylanoides. Minimum inhibitory concentration determinations were made using the National Committee for Clinical Laboratory Standards reference broth microdilution method. Variation in the rank order and frequency of the different species of Candida was observed over time and by geographic area. The proportion of BSI due to C. albicans and C. glabrata increased and C. parapsilosis decreased over time in Canada, the USA and Europe. C. glabrata was an infrequent cause of BSI in Latin America and the Asia-Pacific region. Very little variation in fluconazole susceptibility was observed among isolates of C. albicans, C. tropicalis and C. parapsilosis. These species accounted for 78% of all BSI and remained highly susceptible (91-100% susceptible) to fluconazole from 1992 to 2001 irrespective of geographic origin. The prevalence of fluconazole resistance among C. glabrata isolates was variable both over time and among the various countries and regions. Resistance to fluconazole among C. glabrata isolates was greatest in the USA and varied by US census region (range 0-23%). These observations are generally encouraging relative to the sustained usefulness of fluconazole as a systemically active antifungal agent for the treatment of candida BSI. PMID- 14748800 TI - New clinical presentations of invasive aspergillosis in non-conventional hosts. AB - Infections by Aspergillus spp. are most typically associated with invasive pulmonary aspergillosis. However, an increasing number of reports deal with unusual manifestations of invasive aspergillosis. In the lung this may take the form of chronic invasive pulmonary aspergillosis, bronchocentric granulomatosis or tracehobronchitis. A number of extrapulmonary infections have been noted, sometimes in immunocompetent individuals. Examples include vertebral osteomyelitis, primary cutaneous aspergillosis (such as in premature neonates), prosthetic vascular graft infection and infective endocarditis. Early recognition of these entities, prompt initiation of new, highly active antifungal therapies and adjunctive surgical management may improve the prognosis of these conditions. PMID- 14748801 TI - Mucormycosis and entomophthoramycosis: a review of the clinical manifestations, diagnosis and treatment. AB - The class Zygomycetes is divided into two orders, Mucorales and Entomophthorales. These two orders produce dramatically different infections. Genera from the order Mucorales (Rhizopus, Mucor, Rhizomucor, Absidia, Apophysomyces, Cunninghamella and Saksenaea) cause an angioinvasive infection called mucormycosis. Mucormycosis presents with rhino-orbito-cerebral, pulmonary, disseminated, cutaneous, or gastrointestinal involvement. Immunocompromising states such as haematological malignancy, bone marrow or peripheral blood stem cell transplantation, neutropenia, solid organ transplantation, diabetes mellitus with or without ketoacidosis, corticosteroids, and deferoxamine therapy for iron overload predispose patients to infection. Mucormycosis in immunocompetent hosts is rare, and is often related to trauma. Mortality rates can approach 100% depending on the patient's underlying disease and form of mucormycosis. Early diagnosis, along with treatment of the underlying medical condition, surgery, and an amphotericin B product are needed for a successful outcome. Genera from the order Entomophthorales produce a chronic subcutaneous infection called entomophthoramycosis in immunocompetent patients. This infection occurs in tropical and subtropical climates. The genus Basidiobolus typically produces a chronic subcutaneous infection of the thigh, buttock, and/or trunk. Rarely, it has been reported to involve the gastrointestinal tract. The genus Conidiobolus causes a chronic infection of the nasal submucosa and subcutaneous tissue of the nose and face. This paper will review the clinical manifestations, diagnosis and treatment of mucormycosis and entomophthoramycosis. PMID- 14748802 TI - Infections due to emerging and uncommon medically important fungal pathogens. AB - The emergence of less common but medically important fungal pathogens contributes to the rate of morbidity and mortality, especially in the increasingly expanding population of immunocompromised patients. These pathogens include septate filamentous fungi (e.g., Fusarium spp., Scedosporium spp., Trichoderma spp.), nonseptate Zygomycetes, the endemic dimorphic pathogen Penicillium marneffei, and non-Cryptococcus, non-Candida pathogenic yeast (e.g., Trichosporon spp.). The medical community is thus called upon to acquire an understanding of the microbiology, epidemiology and pathogenesis of these previously uncommon pathogens in order to become familiar with the options for prevention and treatment. PMID- 14748803 TI - Human fusariosis. AB - Fusarium species frequently implicated in human infections include F. solani, F. oxysporum and F. moniliforme. Among immunocompetent patients, tissue breakdown (as caused by trauma, severe burns or foreign body) is the risk factor for fusariosis. Infections include keratitis, onychomycosis and occasionally peritonitis and cellulitis. Treatment is usually successful and requires removal of the foreign body as well as antifungal therapy. Among immunocompromised patients, mainly patients with haematological malignancies, Fusarium spp. are the second most common pathogenic mould. Risk factors for disseminated fusariosis include severe immunosuppression (neutropenia, lymphopenia, graft-versus-host disease, corticosteroids), colonisation, tissue damage, and receipt of a graft from an HLA-mismatched or unrelated donor. Clinical presentation includes refractory fever (> 90%), skin lesions and sino-pulmonary infections ( approximately 75%). Type of skin lesions includes ecthyma-like, target, and multiple subcutaneous nodules. Skin lesions lead to diagnosis in > 50% of patients and precede fungemia by approximately 5 days. In contrast to disseminated aspergillosis, disseminated fusariosis can be diagnosed by blood cultures in 40% of patients. Histopathology reveals hyaline acute-branching septate hyphae similar to those found in aspergillosis. Mortality from fusarial infections in immunocompromised patients ranges from 50% to 80%. Host immune status is the single most important factor predicting outcome. Persistent neutropenia and corticosteroid therapy significantly affect survival. Optimal treatment has not been established. Anecdotal successes have been reported with various agents (high-dose amphotericin B, lipid-based amphotericin B formulations, itraconazole, voriconazole) and with cytokine-stimulated granulocyte transfusions. Preventing fusariosis relies on detection and treatment of cutaneous damage prior to commencing immunosuppression and decreasing environmental exposure to Fusaria (via air and water). PMID- 14748804 TI - Invasive infections caused by Blastoschizomyces capitatus and Scedosporium spp. AB - Blastoschizomyces capitatus, Scedosporium prolificans and S. apiospermum are emerging fungal pathogens that may cause disseminated disease in neutropenic patients. They can present as fever resistant to antibiotics and to wide-spectrum antifungal agents, although they may involve almost every organ. The proportion of recovery from blood cultures is high and they are characteristically resistant to most antifungal agents. Prognosis is poor unless patients recover from neutropenia. Voriconazole has good in-vitro activity and is currently the drug of choice for these infections. PMID- 14748805 TI - Antifungal prophylaxis with azole derivatives. AB - In recent years, several reports have underlined the increasing role of fungal infections as a cause of morbidity and mortality in hospitalised patients. For this reason, and also in light of the high mortality rate associated with these infections, chemoprophylaxis has been advocated by several authors. The available evidence suggests that both fluconazole and itraconazole are able to decrease candida colonisation and infection, when compared with placebo or with nonabsorbable antifungals. Data seem also to suggest that a decrease in fungus related mortality can be achieved with prophylaxis, although with little effect on overall mortality, probably because of the importance of severe underlying diseases. Itraconazole proved to be effective in the prevention of fungal infections, including invasive aspergillosis, although with increased incidence of side-effects, often leading to treatment discontinuation. The other side of the coin is that antifungal prophylaxis might have untoward effects, such as the selection of triazole-resistant Candida strains or the induction of resistance. In addition, some authors have suggested that the use of triazoles might modulate the pattern of infecting organisms in cancer patients, increasing the risk of both aspergillosis and bacteremia. In conclusion, antifungal prophylaxis with triazole antifungals should be used with caution, only in patients at high risk for invasive fungal infections. These include allogeneic bone marrow transplant patients (especially those with mismatched or unrelated donors), acute myeloid leukaemia patients treated with high-dose cytarabine (C-ara), very-low-birth weight infants, patients with chronic granulomatous disease, and high-risk surgical and intensive-care unit patients. PMID- 14748806 TI - Oral versus intravenous therapy in the treatment of systemic mycosis. AB - The great majority of systemic fungal infections require long-term therapy that often extends 6-12 months, particularly in immunosuppressed patients. It can be difficult to comply with this requirement when the drug to be used is only available for intravenous administration, because problems related to maintaining a permeable venous pathway for long periods arise. The availability of an intravenously (IV) and orally (PO) administered drug can solve this problem by making sequential therapy possible. Voriconazole is a new antifungal agent that, apart from satisfying this requirement because it has a high oral bioavailability, presents a broad spectrum of antifungal activity that makes its use possible, a priori, in the initial and/or sequential IV/PO treatment of any systemic mycotic infection. Based on current costs there is potential for savings compared with liposomal amphotericin B. PMID- 14748807 TI - Voriconazole-a new therapeutic agent with an extended spectrum of antifungal activity. AB - Voriconazole is a new antifungal agent that can be given orally and intravenously. It has proven efficacy for treating candidosis and invasive aspergillosis as well as other mould infections, such as those caused by Fusarium and Scedosporium spp. The drug is generally well tolerated. PMID- 14748809 TI - In defence of polypharmacy. PMID- 14748810 TI - Adverse drug reactions in elderly patients. AB - Many studies from around the world show a correlation between increasing age and adverse drug reaction (ADR) rate, at least for some medical conditions. More than 80% of ADRs causing admission or occurring in hospital are type A (dose-related) in nature, and thus predictable from the known pharmacology of the drug and therefore potentially avoidable. Frail elderly patients appear to be particularly at risk of ADRs and this group is also likely to be receiving several medicines. The toxicity of some drug combinations may sometimes be synergistic and be greater than the sum of the risks of toxicity of either agent used alone. In order to recognize and to prevent ADRs (including drug interactions), good communication is crucial, and prescribers should develop an effective therapeutic partnership with the patient and with fellow health professionals. Undergraduate and postgraduate education in evidence-based therapeutics is also vitally important. The use of computer-based decision support systems (CDSS) and electronic prescribing should be encouraged, and when problems do occur, health professionals need to be aware of their professional responsibility to report suspected adverse drug events (ADEs) and ADRs. "Rational" or "obligatory" polypharmacy is becoming a legitimate practice as increasing numbers of individuals live longer and the range of available therapeutic options for many medical conditions increases. The clear risk of ADRs in this situation should be considered in the context that dose-related failure of existing therapy to manage the condition adequately may be one of the most important reasons for admission of the elderly to hospital. Thus, age itself should not be used as a reason for withholding adequate doses of effective therapies. PMID- 14748811 TI - Application of data mining techniques in pharmacovigilance. AB - AIMS: To discuss the potential use of data mining and knowledge discovery in databases for detection of adverse drug events (ADE) in pharmacovigilance. METHODS: A literature search was conducted to identify articles, which contained details of data mining, signal generation or knowledge discovery in relation to adverse drug reactions or pharmacovigilance in medical databases. RESULTS: ADEs are common and result in significant mortality, and despite existing systems drugs have been withdrawn due to ADEs many years after licensing. Knowledge discovery in databases (KDD) is a technique which may be used to detect potential ADEs more efficiently. KDD involves the selection of data variables and databases, data preprocessing, data mining and data interpretation and utilization. Data mining encompasses a number of statistical techniques including cluster analysis, link analysis, deviation detection and disproportionality assessment which can be utilized to determine the presence of and to assess the strength of ADE signals. Currently the only data mining methods to be used in pharmacovigilance are those of disproportionality, such as the Proportional Reporting Ratio and Information Component, which have been used to analyse the UK Yellow Card Scheme spontaneous reporting database and the WHO Uppsala Monitoring Centre database. The association of pericarditis with practolol but not with other beta-blockers, the association of captopril and other angiotensin converting enzymes with cough, and the association of terfenadine with heart rate and rhythm disorders could be identified by mining the WHO database. CONCLUSION: In view of the importance of ADEs and the development of massive data storage systems and powerful computer systems, the use of data mining techniques in knowledge discovery in medical databases is likely to be of increasing importance in the process of pharmacovigilance as they are likely to be able to detect signals earlier than using current methods. PMID- 14748812 TI - Comparative population pharmacokinetics of lorazepam and midazolam during long term continuous infusion in critically ill patients. AB - AIMS: It is well established that there is a wide intra- and interindividual variability in dose requirements for lorazepam and midazolam in intensive care patients. The objective of this study was to compare the population pharmacokinetics of lorazepam and midazolam after long-term continuous infusion in mechanically ventilated critically ill patients. METHODS: Forty-nine critically ill patients randomly received either lorazepam (n = 28) or midazolam (n = 21) by continuous infusion for at least 24 h. Multiple blood samples were obtained for determination of the drug and metabolite concentrations by HPLC. Population pharmacokinetic models were developed using the Non-Linear Mixed Effect Modelling (NONMEM) program. The influence of selected covariates was investigated. The prospective performance of the models was evaluated on the basis of results in separate groups of patients for lorazepam (n = 31) and midazolam (n = 33). RESULTS: The pharmacokinetics of lorazepam were best described by a two-compartment model. Alcohol abuse, positive end expiratory pressure (PEEP) and age were identified as significant covariates. Total body clearance for patients without alcohol abuse was 4.13 - (PEEP - 5) x 0.42 l h-1, and 0.74 l h-1 for patients with alcohol abuse. The volume of distribution was 0.74 l, the steady state volume of distribution was 56 - (age - 58) x 2.1 l and the intercompartmental clearance was 10 l h-1. The proportional residual error was 15% and the median absolute prediction error was 13.6% with a bias of 1.5%. The pharmacokinetics of midazolam were best described by a two-compartment model with alcohol abuse, APACHE score and age as significant covariates. Total body clearance for patients without alcohol abuse was 11.3 - (age - 57) x 0.14 l h-1, and 7.27 - (age -57) x 0.14 l h-1 for patients with alcohol abuse. The volume of distribution was 7.15 l, the steady state volume of distribution was 431 l, and the intercompartmental clearance was 40.8 - (APACHE score - 26) x 2.75 l h-1. The proportional residual error was 31% with an additive residual error of 32 ng ml 1. The median absolute prediction error was 12.9% with a bias of 1.2%. The prospective performance in the lorazepam evaluation group was better with the covariate adjusted model, but in the midazolam evaluation group it was not better than with the simple model. In all models a tendency to overestimate the lower plasma concentrations was observed. CONCLUSIONS: The pharmacokinetics of both lorazepam and midazolam were well described by a two-compartment model. Inclusion of alcohol abuse and age as covariates improved both models. PEEP was identified as an additional covariate for lorazepam, and the APACHE score for midazolam. For both drugs there is a large interindividual variability in their pharmacokinetics when used for long-term sedation in critically ill patients. However, the intra individual variability is much lower for lorazepam. PMID- 14748813 TI - Population pharmacokinetics of artemether and dihydroartemisinin following single intramuscular dosing of artemether in African children with severe falciparum malaria. AB - AIMS: To determine the population pharmacokinetics of artemether and dihydroartemisinin in African children with severe malaria and acidosis associated with respiratory distress following an intramuscular injection of artemether. METHODS: Following a single intramuscular (i.m.) injection of 3.2 mg kg-1 artemether, blood samples were withdrawn at various times over 24 h after the dose. Plasma was assayed for artemether and dihydroartemisinin by gas chromatography-mass spectrometry. The software program NONMEM was used to fit the concentration-time data and investigate the influence of a range of clinical characteristics (respiratory distress and metabolic acidosis, demographic features and disease) on the pharmacokinetics of artemether and dihydroartemisinin. RESULTS: A total of 100 children with a median age of 36.4 (range 5-108) months were recruited into the study and data from 90 of these children (30 with respiratory distress and 60 with no respiratory distress) were used in the population pharmacokinetic analysis. The best model to describe the disposition of artemether was a one-compartment model with first-order absorption and elimination. The population estimate of clearance (clearance/bioavailability, CL/F) was 14.3 l h-1 with 53% intersubject variability and that of the terminal half-life was 18.5 h. If it was assumed that artemisin displays "flip-flop" kinetics, the elimination half-life was estimated to be 21 min and the corresponding volume of distribution was 8.44 l, with an intersubject variability of 104%. None of the covariates could be identified as having any influence on the disposition of artemether. The disposition of dihydroartemisinin was fitted separately using a one-compartment linear model in which the volume of distribution was fixed to the same value as that of artemether. Assuming that artemether is completely converted to dihydroartemisinin, the estimated value of CL/F for dihydroartemisinin was 93.5 l h-1, with an intersubject variability of 90.2%. The clearance of dihydroartemisinin was formation rate limited. CONCLUSIONS: Administration of a single 3.2 mg kg-1 i.m. dose of artemether to African children with severe malaria and acidosis is characterized by variable absorption kinetics, probably related to drug formulation characteristics rather than to pathophysiological factors. Use of i.m. artemether in such children needs to be reconsidered. PMID- 14748814 TI - Modelling acute tolerance to the EEG effect of two benzodiazepines. AB - AIMS: We studied the development of acute tolerance to the EEG effect of midazolam and the new benzodiazepine Ro 48-6791. METHODS: Nine young (24-28 years) and nine elderly (67-81 years) male volunteers received midazolam and Ro 48-6791 computer-controlled, targeting linearly increasing plasma concentrations for 30 min (targeted slopes: 40 and 20 ng ml-1 min-1 for midazolam, 3 and 1.5 ng ml-1 min-1 for Ro 48-6791, for young and elderly, respectively) and a constant concentration for the following 15 min. After recovery, the same infusion scheme was repeated. Plasma concentrations of midazolam, Ro 48-6791 and its metabolite Ro 48-6792 were determined from arterial blood samples. The hypnotic effect was assessed using the median frequency of the EEG power spectrum. RESULTS: The concentration-effect relationship in each infusion cycle could be described by a sigmoid Emax model. The half-maximum concentration EC50 was higher in the second infusion cycle compared with the first one (midazolam, 47% (2.3-91.6%) and 37% (5.3-69.5%); Ro 48-6791, 22% (-2.8% to 44.6%) and 43% (3.4-82.4%) for young and elderly; mean and 95% confidence interval). The complete time course of the EEG median frequency could be described by an interaction between the parent drug in an effect compartment and a hypothetical competitive drug in an additional tolerance compartment. For Ro 48-6791, the use of its metabolite Ro 48-6792 as competitive compound also gave appropriate results. CONCLUSION: Midzolam and Ro 48-6791 showed acute tolerance to the EEG effect which might be caused by competitive interaction with the metabolite. PMID- 14748815 TI - Pharmacokinetic and pharmacodynamic assessment of a five-probe metabolic cocktail for CYPs 1A2, 3A4, 2C9, 2D6 and 2E1. AB - AIMS: The primary objectives of the present study were to establish whether there was a pharmacokinetic or pharmacodynamic interaction between the probe drugs caffeine (CYP1A2), tolbutamide (CYP2C9), debrisoquine (CYP2D6), chlorzoxazone (CYP2E1) and midazolam (CYP3A4), when administered in combination as a cocktail. Furthermore, the tolerability of these probe drugs, both alone and in combination as a cocktail was assessed. METHODS: Twelve healthy volunteer subjects (age range 22-48 years) were entered into an open, fixed sequence, 6-limb, single centre study. The randomization was such that all drugs were given individually followed by the full "cocktail" as the last treatment limb. The phenotypic index used to assess the intrinsic activity of the CYP isoforms included metabolite/parent ratios in plasma and urine (CYPs 1A2, 2E1 & 2C9), parent/metabolite ratios in urine (CYP2D6) and plasma AUClast (CYP3A4). Blood pressure and blood glucose measurements were used to assess pharmacodynamic interactions. Tolerability was assessed through reporting of adverse events RESULTS: Overall, there was little evidence that the probe drugs interacted metabolically when co-administered as the cocktail. The ratio of the geometric mean (and 90% confidence interval) of the phenotypic index, obtained after administration of the probe as part of the cocktail and when given alone were: caffeine, 0.86 (0.67-1.10), midazolam, 0.96 (0.74-1.24), tolbutamide, 0.86 (0.72-1.03), debrisoquine 1.04 (0.97-1.12) and chlorzoxazone, 0.95 (0.86-1.05). There was no difference in blood pressure and blood glucose concentrations following the cocktail and dosing of the individual probes. There was no effect on ECG recordings at any time-point. The adverse events reported for individual drug administrations were mild, transient and expected. Overall no more adverse events were reported on the cocktail study days than on the days when the drugs were administered alone. CONCLUSIONS: The five probe drugs when coadministered, in this dosing regimen, demonstrated no evidence of either a metabolic or pharmacodynamic interaction that might confound the conclusions drawn during a cocktail study. The present cocktail methodology has the potential to become a useful tool to aid the detection of clinically important drug-drug interactions during drug development. PMID- 14748816 TI - Assessment of the relative in vivo potency of the hydroxylated metabolite of darifenacin in its ability to decrease salivary flow using pooled population pharmacokinetic-pharmacodynamic data. AB - AIMS: To describe the population pharmacokinetic-pharmacodynamic relationship between darifenacin (UK-88,525) and its hydroxylated metabolite (UK-148,993), and the reduction in salivary flow (SF, a M3-mediated response). This enabled an estimation of the in vivo potency of the metabolite to decrease SF relative to that of the parent drug. METHODS: A total of 262 individuals were pooled from 11 Phase 1 studies and one Phase 2 study. A comparison was made between a series of pharmacodynamic models (direct-effect, indirect-effect, link and binding model) using NONMEM. RESULTS: The binding model yielded the best description of the decrease in SF by fully accounting for the time course of the pharmacodynamic effect. An internal validation exercise demonstrated the robustness of this model. Covariate analysis identified a circadian rhythm in SF. This model, with confidence intervals (CI) determined by likelihood profiling, indicated that the relative potency of the metabolite to darifenacin to reduce SF was 11.1% (95% CI 3.8, 19.6). This implied that the metabolite was ninefold less potent than darifenacin in vivo. Accounting for the unbound fraction of darifenacin (2%) and its metabolite (13%), the in vivo protein binding-corrected relative potency was estimated to be 2.1%, indicating that the metabolite was 50-fold less potent than the parent drug. The model supported the assumption that no other metabolites contributing to the impairment of the SF were formed during first-pass, and that the development of sensitization or tolerance was not evident over time. The validation process indicated that the i.v.-oral crossover study was necessary for the estimation of the relative potency. CONCLUSIONS: Population modelling of darifenacin and its hydroxylated metabolite yielded individual pharmacokinetic predictions that could be used to assess the in vivo potency of the metabolite to decrease SF relative to that of the parent drug. The metabolite had a negligible effect on SF. PMID- 14748817 TI - Effect of rifampicin on pravastatin pharmacokinetics in healthy subjects. AB - AIMS: Previous work has shown that rifampicin, a potent inducer of several cytochrome P450 (CYP) enzymes and transporters, decreased the plasma concentrations of simvastatin acid by more than 90%. This study was conducted to investigate the effect of rifampicin on the pharmacokinetics of pravastatin. METHODS: In a randomised, cross-over two-phase study with a washout of 4 weeks, 10 healthy volunteers received a 5-day pretreatment with rifampicin (600 mg daily) or placebo. On day 6, a single 40 mg dose of pravastatin was administered orally. Plasma concentrations of pravastatin were measured up to 12 h by a sensitive LC-MS-MS method. RESULTS: During the rifampicin phase, the mean total area under the plasma concentration-time curve of pravastatin [AUC(0-infinity )] was 69% (range 24-220%) of the corresponding value during the placebo phase (P < 0.05, 95% confidence interval for the difference -51.9 - -0.4 ng ml-1.h). In five of the 10 subjects the AUC(0-infinity ) of pravastatin during the rifampicin phase was 50% or less of that during the placebo phase. Rifampicin had no significant effect on the peak concentration, elimination half-life or renal clearance of pravastatin. CONCLUSIONS: Rifampicin caused a statistically significant decrease in the plasma concentration of pravastatin given as a single oral dose to healthy subjects. However, the effect of rifampicin varied greatly between subjects. The mean rifampicin-induced decrease in pravastatin concentration was considerably smaller than that observed previously for simvastatin. PMID- 14748818 TI - Cardiovascular prophylaxis with aspirin: costs of supply and management of upper gastrointestinal and renal toxicity. AB - AIMS: To determine the cost to the NHS of prescribed low-dose aspirin. METHODS: This was a population based observational cohort study. Patients from Tayside Scotland (17 244 new users of dispensed aspirin each with 10 matched comparators) were included. A pragmatic analysis totalled costs from the start to end of the study and compared these with a matched cohort of aspirin nonusers to estimate excess costs. Fastidious analyses were done of subjects with no prior history of upper gastrointestinal (UGI) or renal disease where the cost that occurred during aspirin exposure, the 30 days following aspirin exposure and subsequent nonexposure was calculated adjusting for risk factors in each period. RESULTS: Subjects took aspirin for only 1.18 of the 2.53 years follow-up (47% compliance). Aspirin use cost an additional 49.86 UK pounds per year (pragmatic analysis) made up of 1.96 UK pounds for aspirin tablets (4%), 5.49 UK pounds for dispensing costs (11%), 24.60 UK pounds for UGI complications (49%) and 17.81 UK pounds for renal complications (36%). The costs for managing complications were substantially lower in the fastidious analysis (2.66 UK pounds for UGI complications and 2.92 UK pounds for renal complications). Assuming that the antiplatelet trial meta-analysis is an accurate assessment of the benefits of aspirin, the costs of preventing one vascular event lay between 62 500 UK pounds (primary prevention, pragmatic analysis) and 867 UK pounds (secondary prevention, fastidious analysis). These costs may be underestimates due to the low compliance observed. CONCLUSIONS: Compliance with aspirin was poor. Serious adverse events were uncommon but despite this aspirin cost the NHS between 6 and 25 times the cost of aspirin tablets due to dispensing costs and the cost of managing adverse effects. PMID- 14748819 TI - The influence of diet upon liver function tests and serum lipids in healthy male volunteers resident in a Phase I unit. AB - AIM: To investigate the effect of diet upon liver function tests and serum lipids within the restricted environment of a Phase I unit. METHODS: An open randomized three-way crossover study was designed with subjects consuming three types of diet. The diets comprised, a balanced normal calorie diet, a high-carbohydrate high-calorie diet and a high-fat high-calorie diet. Each diet was consumed in a randomized sequence over 8 days with a recovery period of 14 days between periods. The blood concentrations of various laboratory parameters were measured at intervals throughout each dietary period and during the recovery periods. RESULTS: Blood transaminase activity and triglyceride concentrations increased significantly whilst subjects consumed a high-carbohydrate high-calorie diet but not when fed either a high-fat high-calorie diet or a balanced normal calorie diet. CONCLUSIONS: The rises in transaminases and triglycerides were caused by the carbohydrate content of the diet rather than its calorific value. Sucrose rather than starch was the carbohydrate which caused the rise in transaminases and triglycerides. The importance of controlling diet in Phase I studies is stressed. PMID- 14748820 TI - Sildenafil does not improve nitric oxide-mediated endothelium-dependent vascular responses in smokers. AB - AIMS: To examine the hypothesis that sildenafil, a phosphodiesterase type 5 inhibitor that inhibits cGMP breakdown, could enhance nitric oxide-mediated vasodilation and reverse endothelial dysfunction in chronic smokers. METHODS: Flow-mediated dilation of the brachial artery and forearm postischemic reactive hyperemia (both nitric oxide-mediated responses) were measured before and after sildenafil 50 mg and placebo in a double-blind, randomized, crossover study in 9 men who were chronic smokers (21 +/- 3 pack years). RESULTS: There was no significant change in flow-mediated dilation after either sildenafil (0.18%, 95%CI -1.7-2%) or placebo (0.24%, 95%CI -2.8-3.3%) (P = 0.88 and 0.8, respectively). Sildenafil had no significant effect on resting forearm blood flow or postischemic reactive hyperemia (P = 0.39 and 0.7, respectively). Resting heart rate and blood pressure were unaffected by sildenafil. CONCLUSIONS: Acute sildenafil administration did not improve endothelial function in chronic smoking men. PMID- 14748821 TI - Protective role of glutathione S-transferase P1 (GSTP1) Val105Val genotype in patients with bronchial asthma. AB - BACKGROUND: Glutathione S-transferase P1 (GSTP1), the abundant isoform of glutathione S-transferases (GSTs) in lung epithelium, plays an important role in cellular protection against oxidative stress and toxic foreign chemicals. It has been suggested that polymorphisms in the GSTP1 gene are associated with asthma and related phenotypes. As significant interindividual and interethnic differences exist in the distribution of xenobiotic-metabolizing enzymes, we have studied the GSTP1 Ile105Val polymorphism in patients with asthma in a Turkish sample. METHODS: GSTP1 Ile105Val polymorphism in exon 5 was determined in 210 patients with asthma (112 extrinsic and 108 intrinsic) and 265 control individuals without lung diseases and without history of allergy or atopy, using polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) techniques. RESULTS: The proportion of GSTP1 Val105 homozygotes was significantly lower in the patients with asthma than in the control individuals (3.8% vs 12.1%). The odds ratio for GSTP1 Val105 homozygotes vs all other genotypes was 0.29 (95%CL 0.13-0.64, p = 0.01) for asthmatics. The distribution of GSTP1 Ile105Val genotypes and the frequency of GSTP1 Val105Val homozygotes (3.7% vs 3.9%) was not significantly different between extrinsic and intrinsic asthmatics. CONCLUSION: These results suggest a significant association between GSTP1 Ile105Val polymorphism and susceptibility to asthma and that the GSTP1 Val105Val genotype may be protective against developing this disease. PMID- 14748822 TI - Effect of food on the relative bioavailability of two oral formulations of posaconazole in healthy adults. AB - AIMS: This randomized, crossover, single-dose study evaluated the relative oral bioavailability of posaconazole suspension and coprecipitate tablet formulations. Additionally, the study determined whether systemic exposure to posaconazole was affected by prandial status or by the fat content of a meal. METHODS: This was a randomized, open-label, four-way crossover, single-dose study in 20 healthy men. Posaconazole pharmacokinetics were evaluated over 72 h following a single oral dose of posaconazole suspension (200 mg/5 ml) administered with a high-fat meal, a nonfat breakfast, or after a 10 h fast, or posaconazole tablets (2 x 100 mg) administered with a high-fat meal. RESULTS: The posaconazole suspension showed a significant increase in bioavailability compared with the tablet (increase in AUC(0,72 h) = 137% (90% confidence interval (CI) 119%, 156% and Cmax = 123% (90% CI 104%, 146%). The mean increases in AUC(0,72 h) and Cmax values were about 400% when administered with a high-fat meal compared with administration of the suspension in the fasting state (AUC(0,72 h) 90% CI 343%, 448%; Cmax 90% CI 352%, 493%). Administration of the suspension with a nonfat meal enhanced exposure, resulting in an increase in AUC(0,72 h) of 264% (90% CI 231%, 302%) and in Cmax of 296% (90% CI 250%, 350%) relative to the fasted state. CONCLUSIONS: The suspension formulation of posaconazole was associated with enhanced systemic exposure and increased relative bioavailability compared with the tablet. Food substantially enhanced the rate and extent of posaconazole absorption in healthy subjects. PMID- 14748823 TI - Cerebrospinal fluid pharmacokinetics of paracetamol after intravenous propacetamol in a former preterm infant. PMID- 14748826 TI - Isolation by language and distance in Belgium. AB - The isonymy structure of trilingual Belgium was studied using the surname distributions for 1,118,004 private telephone users. The users were distributed in 77 Flemish, 76 French, and 3 German speaking towns, selected on a geographic basis to form an approximately regular grid over Belgium. Lasker's distance was found to be considerably higher between languages than within languages. For the whole of Belgium, irrespective of language, it was highly correlated with linear geographic distance, with r = 0.721+/-0.014, which is the highest correlation observed in European countries to date. Within Belgium and within languages, the correlation was highest among the Flemish (r = 0.878 +/- 0.007), and lowest among the French (r = 0.631+/-0.020). Isolation by distance in Belgium is the highest we have found in Europe, and as high as in Switzerland where the different languages are separated by geographical barriers. This is not the case in Belgium, so that the considerable isolating power of languages emerges clearly from the present analysis. From the comparison of Lasker's distance between (9.48) and within (8.16) languages, and from its regression over geographic distance (b = 0.01206), it was possible to establish a quantitative relationship between the isolating power of languages and that of geographic distance as (9.48 8.16)/0.01206 = 109 kilometres. This transformation of language distance into an equivalent geographic distance, given here for Belgium, can be applied to any similar geo-linguistic situation. PMID- 14748827 TI - Molecular analysis of myophosphorylase deficiency in Dutch patients with McArdle's disease. AB - We report on 8 Dutch patients with McArdle's disease from 6 unrelated families. Molecular analysis revealed the presence of four previously described mutations: the common R49X mutation, the IVS14+1G>A mutation and the recently reported R269X and Y84X nonsense mutations; and two new molecular defects: a missense mutation R138W in the homozygous state in two siblings, and a frameshift mutation c.1797delT. This first genetic study of patients from The Netherlands with McArdle's disease confirms that the R49X mutation is also the most common in Dutch patients, and that there is genetic heterogeneity within this population. Moreover, our data support the hypothesis that the Y84X mutation is a relatively frequent mutation in McArdle's patients with a Central European background, and expand the already crowded map of mutations within the PYGM gene responsible for McArdle's disease. PMID- 14748828 TI - Mitochondrial DNA sequence diversity in a sedentary population from Egypt. AB - The mitochondrial DNA (mtDNA) diversity of 58 individuals from Upper Egypt, more than half (34 individuals) from Gurna, whose population has an ancient cultural history, were studied by sequencing the control-region and screening diagnostic RFLP markers. This sedentary population presented similarities to the Ethiopian population by the L1 and L2 macrohaplogroup frequency (20.6%), by the West Eurasian component (defined by haplogroups H to K and T to X) and particularly by a high frequency (17.6%) of haplogroup M1. We statistically and phylogenetically analysed and compared the Gurna population with other Egyptian, Near East and sub Saharan Africa populations; AMOVA and Minimum Spanning Network analysis showed that the Gurna population was not isolated from neighbouring populations. Our results suggest that the Gurna population has conserved the trace of an ancestral genetic structure from an ancestral East African population, characterized by a high M1 haplogroup frequency. The current structure of the Egyptian population may be the result of further influence of neighbouring populations on this ancestral population. PMID- 14748829 TI - Pattern of mtDNA variation in three populations from Sao Tome e Principe. AB - We have analysed the matrilineal genetic composition of three self-reported ethnic groups from Sao Tome e Principe (Gulf of Guinea), an African archipelago whose settlement begun in the late fifteenth century. Sequence data from the hypervariable segments I (HVS-I) and II (HVS-II) were obtained for 30 Angolares, 35 Forros and 38 Tongas. The repertory of mtDNA lineages in Sao Tome e Principe denoted a fully African maternal pool, primarily arisen from a Central/Southwestern substratum. The absence of any lineages of putative European descent means that the European impact at the mitochondrial pool was virtually nil. Angolares showed a clear reduction of mtDNA diversity and a slight genetic differentiation relative to Tongas or Forros, whereas the latter two groups did not present any signs of genetic boundaries between each other. The data obtained here reinforce the depiction of genetic substructuring in Sao Tome e Principe previously derived from Y-chromosome STRs. In addition, the crossing of mtDNA and Y-STR information led to the inference that the female mediated gene flow within the archipelago was less restricted than the male, a pattern that could be framed in the cultural traditions and socio-historical interactions among the groups. PMID- 14748830 TI - Tests of gene-environment interaction for case-parent triads with general environmental exposures. AB - As knowledge of the human genome continues to grow, more progress is being made towards not only identifying the genes involved in disease susceptibility but also in defining the synergistic role genes play with environmental exposures. The detection of gene-environment interactions is important as it can offer clinicians a potential means of intervention. The discovery of interactions relies heavily on powerful statistical methods. We present a test, FBAT-I, that can be used to investigate gene-environment interaction. The test uses the case parent triad design and protects the statistical inference from potential spurious results due to population admixture. PMID- 14748831 TI - On the power of affected relative pair designs for linkage studies. AB - Using the recurrence risk ratio (lambda), Risch (1990b) indicated that affected pairs of distant relatives are preferable to affected sib-pairs for linkage analysis when lambda is large and the mode of inheritance is additive. By using the optimum test for affected sib-pairs instead of the test used by Risch (1990b), the range of values of lambda for which the affected sib-pair design has larger or smaller power than other pairs is clarified. Risch's conclusion remains true when lambdaO > 2.5, however, sib-pairs have larger power for lower values. As affected sib-pairs occur smore frequently than other relative pairs, when ascertainment costs are non-negligible, they may be the most cost-effective relative pairs to use. PMID- 14748832 TI - Meta-analysis of linkage studies for complex diseases: an overview of methods and a simulation study. AB - Linkage genome scans for complex diseases have low power with the usual sample sizes, and hence meta-analysis of several scans for the same disease might be a promising approach. Appropriate data are now becoming accessible. Here we give an overview of the available statistical methods and current applications. In a simulation study, we compare the power of different methods to combine multipoint linkage scores, namely Fisher's p-value combination, the truncated product method, the Genome Search Meta-Analysis (GSMA) method and our weighting methods. In particular, we investigate the effects of heterogeneity introduced by different genetic marker sets and sample sizes between genome scans. The weighting methods explicitly take those differences into account and have more power in the simulated scenarios than the other methods. PMID- 14748833 TI - Partial purification of mare early pregnancy factor. AB - PROBLEM: Early pregnancy factor (EPF) is an immunosuppressive protein detected in the serum in early pregnancy. We have already reported the development of the rosette inhibition test for mare EPF and have detected EPF in thoroughbreds and ponies. Here, we attempted to purify equine EPF from pregnant mare serum. METHODS OF STUDY: Mare EPF was purified by ultrafiltration and ion-exchange chromatography. Purified EPF was analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), immunoblotting and a neutralization test. EPF activity was estimated as the rosette inhibition titer (RIT) by the rosette inhibition test. RESULTS: Purified EPF bound to carboxymethyl (CM) sepharose and did not adsorb to diethylaminoethyl (DEAE) sepharose. SDS-PAGE revealed that in the final purified fraction there were many proteins. In the immunoblotting analysis, a protein band of 25.8 kDa was detected as the pregnancy-specific band. Further, antibody gained from the 20 to 30 kDa protein band of the final purified fraction neutralized the RIT activity of pregnant mare serum. CONCLUSIONS: Mare EPF was detected in the final purified fraction and had a molecular weight of 25.8 kDa. EPF in the mare is similar to that obtained from the serum of pregnant cows. PMID- 14748834 TI - Early pregnancy factor in cervical mucus of pregnant women. AB - PROBLEM: This study was carried out to determine the presence of early pregnancy factor (EPF) activity in cervical mucus of pregnant women and explore the relationship between EPF activity in sera suggested it and in cervical mucus. METHOD OF STUDY: Cervical mucus and sera were collected from 60 pregnant women in different terms and 25 non-pregnant women, respectively. EPF activity was detected by rosette inhibition assay. RESULTS: EPF activity did exist in cervical mucus of pregnant women and was similar both in cervical mucus and sera. CONCLUSIONS: We report here for the first time the EPF activity in cervical mucus of pregnant women. The parallel change observed in sera suggested it might be another useful index in evaluating embryo viability. PMID- 14748835 TI - Studies with Synthetic Peptides of 80 kDa Human Sperm Antigen (80 kDa HSA). AB - PROBLEM: The 80 kDa human sperm antigen (HSA) is a sperm-specific and conserved antigen, capable of inducing immunological infertility. Partial N-terminal amino acid sequences of 80 kDa HSA (Peptide NT) and its peptides obtained by digestion with endoproteinase Lys-C (peptides 1-4) and endoproteinase Glu-C (peptides 5-6) did not show any sequence homology with reported known proteins deposited in the Gen-Bank. These sequenced peptides were synthesized and conjugated to key hole limpet haemocyanin (KLH) and evaluated for its antifertility effects. The present communication describes the characterization of these peptides and their antibodies. METHOD OF STUDY: Peptides NT, 1, 2, 3 and 4 were synthesized and conjugated to KLH. Antibodies to KLH conjugated peptides were raised in rabbits by active immunization and the antibody titer was determined by enzyme-linked immunosorbent assay (ELISA) using sperm extract coated wells. The binding specificity of the synthetic peptides or purified 80 kDa HSA to their antibodies was assessed in the presence of various doses of respective synthetic peptides or 80 kDa HSA. The binding specificity was further confirmed by Western blot analysis. Antipeptide antibodies were also checked for sperm agglutinating activity, in-vitro. RESULTS: Active immunization of rabbits elicited significant antibody titers against the synthetic peptides, except for peptide 3. Antipeptide antibodies specifically recognized the native protein in an ELISA and induced in vitro agglutination of human, rat and monkey sperm. In addition, Western blot analysis showed that these antipeptide antibodies specifically bind to the 80 kDa HSA band of the sperm extract. CONCLUSION: Synthetic peptides of 80 kDa HSA are immunogenic and antibodies raised against these peptides recognize the native protein detected by ELISA, Western blot analysis. In addition, they possess sperm agglutinating activity. These findings suggest that they are promising candidates in the development of immunocontraceptives. PMID- 14748836 TI - Immunohistochemical localization of the growth hormone in human endometrium and decidua. AB - PROBLEM: Recent evidence of growth hormone (GH) receptor expression in rat endometrium and human myometrium have focused our attention on the role of the GH in endometrial development. We tested the expression of GH in the human endometrium throughout the menstrual cycle and during pregnancy. METHOD OF STUDY: Immunohistochemical study was performed on endometrial specimens of fertile women in different periods of the menstrual cycle and in decidua of pregnant women. RESULTS: Glandular cells of the human endometrium were positive for GH in the mid and late luteal phase. Furthermore, the glandular cells of decidua showed intense staining for GH, while the stromal cells were negative. No immunostaining was expressed in the proliferative or early luteal phase. The intensity levels of staining for GH in decidual specimens were significantly higher than in glandular cells of secretory endometrium specimens (P < 0.01). CONCLUSIONS: The glandular cells of the human endometrium express GH from the late luteal phase throughout pregnancy in the decidual tissue. We suppose that GH plays an important role in blastocyst implantation. PMID- 14748837 TI - Macrophages of human first trimester decidua express markers associated to alternative activation. AB - PROBLEM: Depending on the type of their activation, macrophages may promote TH1- or TH2-type of immune responses. To date, not much is known about the activation phenotype of decidua macrophages, which - together with NK cells - constitute the majority of bone marrow derived cells at this location. METHOD OF STUDY: The study was based on analysis of healthy first trimester decidua by immunohistochemistry and flow cytometry. We analyzed expression of markers characteristic for alternatively activated macrophages (Mphi2). RESULTS: The markers MS-1 (stabilin-1) and coagulation factor XIIIa were found expressed in the interior of decidua macrophages (DMphi). In contrast, indoleamine 2,3 dioxygenase (IDO), an enzyme induced in macrophages by IFNgamma, was not present in DMphi. CONCLUSIONS: First trimester DMphi display phenotypic markers associated to alternatively activated macrophages. In addition, absence of IDO indicates that DMphi are not under a predominant influence of IFNgamma. PMID- 14748838 TI - The influence of extracellular matrix proteins on T-cell proliferation and apoptosis in women with endometriosis or uterine leiomyoma. AB - PROBLEM: Interactions between the extracellular matrix (ECM) and peripheral blood T cells in women with endometriosis and leiomyoma are hardly unknown. We have investigated the influence of two major ECM components, collagen IV (C-IV) and fibronectin (Fn), on T-cell proliferation and apoptosis in women with endometriosis and uterine leiomyoma. beta1 integrin expression, responsible for interactions with ECM proteins, was also studied. METHOD OF STUDY: Peripheral blood lymphocytes were obtained from 53 women (17 with uterine leiomyomas, 18 with endometriosis, and 18 from healthy donors). T cells were exposed to ECM proteins co-immobilized with monoclonal antibody anti-CD3 for 72 hr. Apoptosis and S phase of the cell cycle of the T cells were studied by DNA analysis using flow cytometry. The proliferation of T cells was evaluated by MTT assay. The percentage of CD3+ cells expressing CD29 (beta1 integrin chain) was evaluated by double-color flow cytometry. Results were analyzed statistically using the Mann Whitney test. RESULTS AND CONCLUSIONS: (1) A general increase in the percentage of T cells in S phase could be seen in women with endometriosis and uterine leiomyoma in all culture conditions what may suggest general activation of T cells. (2) A significant increase in the percentage of cells in S phase was shown only in the case of T cells exposed to anti-CD3 + C-IV in both women with uterine leiomyoma and endometriosis. (3) However, no apoptotic cells were observed. (4) T cells from patients with uterine leiomyoma exhibited significantly increased level of proliferation after culture with anti-CD3 + C-IV. (5) More T cells expressed beta1 integrin in women with endometriosis or uterine leiomyoma than in healthy donors. Our data may suggest that increased beta1 integrin expression may enhance T-cell-ECM interactions, which may be responsible for the increased proliferation of T cells but not for apoptosis. Therefore, it is possible that interactions of T cells with ECM proteins, especially with C-IV, may contribute to the pathogenesis of endometriosis and uterine leiomyoma. PMID- 14748839 TI - Extracellular Matrix Protein-dependent Apoptosis of T Cells in Women with a History of Recurrent Spontaneous Abortion. AB - PROBLEM: The purpose of the study was to determine the role of T-cell apoptosis in extracellular matrix (ECM) environment in pregnancy maintenance in women with a history of recurrent spontaneous abortion (RSA). METHOD OF STUDY: Thirty-nine non-pregnant women with the history of RSA (anatomic, genetic, endocrine and microbiologic causes were excluded) and 22 healthy women with the previous successful pregnancy outcome were studied. In addition, 21 women with the history of RSA were also studied at the beginning of their next pregnancy. We studied apoptosis of peripheral blood T cells after culture with monoclonal antibody (mAb) OKT-3 alone or with mAb OKT-3 following ECM proteins: collagen IV (C-IV) or fibronectin (Fn). We used Cell Death Detection ELISA for studying cell death in cell population. In addition, apoptotic peripheral blood T cells were identified by annexin V-PE staining protocol using flow cytometry. CD29+ and CD95+ T-cell surface receptors were also analyzed by flow cytometry. RESULTS: The significantly higher values of enrichment factor: mU of the sample (dying/dead cells) per mU of the corresponding control (viable cells) were observed after peripheral blood T-cell culture with C-IV (P = 0.0002) or Fn (P = 0.004) in samples of non-pregnant women with the history of RSA when compared with control women. The significantly higher values of enrichment factor were observed after peripheral blood T-cell culture with C-IV in samples of pregnant women with the history of RSA with successful pregnancy outcome when compared with pregnant women with the history of RSA with failed pregnancy outcome (P = 0.01). However, the percentage of apoptotic T cells stained by annexin V was significantly lower in non-pregnant RSA women compared with control (P = 0.0001). CD95 expression was significantly lower in non-pregnant RSA women compared with control (P = 0.01). CONCLUSIONS: Apoptosis of T cells might be an interesting possible explanation of successful pregnancy outcome in women with the history of RSA. PMID- 14748840 TI - To die or not to die: the function of the transcription factor NF-kappaB in embryos exposed to stress. AB - BACKGROUND: Cytokines operating in the embryo and embryonic microenvironment determine, to a significant extent, whether pregnancy is completed successfully or results in embryonic loss or maldevelopment. They act as activators of specific transcription factors, which control cell responses such as cell proliferation differentiation and apoptosis. One such transcription factor is the nuclear factor-kappaB (NF-kappaB), which is presently seen as a key molecule controlling the apoptosis process. In the light of evidence that a majority of embryopathic stresses, regardless of their nature, first disturb the apoptotic process, it is conceivable, that NF-kappaB may play an important role in regulating the resistance of embryos to embryopathic stresses. In this brief review, we discuss such a possibility based on data characterizing expression and function of NF-kappaB in the embryo and extraembryonic tissues during normal embryogenesis as well as after exposure to various embryopathic stresses. METHODS: Critical review of existing data. RESULTS: Data summarized in this review suggest that (a) practically all NF-kappaB/Rel family members are expressed in embryonic, trophoblast and uterine cells in a developmental stage- and cell type-specific manner; (b) NF-kappaB-mediated anti-apoptotic signaling in embryonic cells seems to be indispensable for proper development during the organogenesis stage, (c) NF-kappaB activity in stress-targeted embryonic and extraembryonic structures directly correlates with their ability to resist stress induced process of embryo loss and maldevelopment. CONCLUSION: Data presented in this review suggest that NF-kappaB may act as a protector of embryos exposed to embryopathic stresses, possibly, because of the ability of NF-kappaB to prevent the induction of programmed cell death as well as to activate cell proliferation. PMID- 14748841 TI - Antiphosphatidylserine antibodies affect rat yolk sacs in culture: a mechanism for fetal loss in antiphospholipid syndrome. AB - PROBLEM: A variety of reproductive impairments have been reported in the context of the antiphospholipid syndrome (APS). APS is associated with the presence of antibodies to negatively charged phospholipids that may affect the outcome of pregnancy. METHOD OF STUDY: Rat embryos were cultured within their yolk sacs. The effects of two antiphosphatidylserine monoclonal aPS antibodies (HL5B, RR7F) regarding their influence on growth and apoptotic events of the yolk sacs, as well as on growth and the morphology of the embryos, were studied. RESULTS: Exposure of rat embryos and their yolk sacs to aPS inhibited yolk sac growth. Moreover, increased number of apoptotic events of giant cells in the aPS-exposed ectoplacental cone was found in comparison with control IgG-exposed giant cells (P < 0.05). No significant damage was observed in the embryos. CONCLUSIONS: The results suggest that aPS affect growth and apoptosis of rat ectoplacental cone. PMID- 14748842 TI - Trophoblasts and soluble adhesion molecules in peripheral blood of women with pregnancy-induced hypertension. AB - PROBLEM: The current hypothesis on the pathogenesis of pregnancy-induced hypertension (PIH) considers it as an endothelial disorder that is first local but with the potential of becoming general. The aim of the work was to investigate the relation of the number of trophoblast cells in maternal peripheral blood against the serum levels of soluble vascular and intercellular cell adhesion molecule (sVCAM-1 and sICAM-1) in PIH. METHOD OF STUDY: Women with PIH were at 28th to 40th week of gestation. Control group were normotensive (NT) pregnant women at 28th to 41st week of gestation. Flow cytometry was used to assess the relative number of the trophoblasts in the peripheral blood. Trophoblasts were labeled with monoclonal anti-human trophoblast protein antibody MCA 277. The presence of sVCAM-1 and sICAM-1 was determined using the enzyme linked immunosorbent assay method. RESULTS: Women with PIH had significantly higher trophoblasts number than NT women (median 19.0, range 5.0-57.0/400 microL versus median 7.0, range 0.0-18.0/400 microL; P = 0.000011) as well as plasma level of sVCAM-1 when compared with NT women (median 730.0, range 325.0-1525.0 ng/mL versus median 493, range 310-1075 ng/mL; P = 0.02). ICAM-1 level in the PIH group was slightly elevated (median 280.0, range 174.0-524.0 ng/mL) when compared with NT women (median 260.0, range 190.0-464.0 ng/mL, P = 0.322). Eight of 21 women with PIH had proteinuria but no correlation was found between this symptom and the laboratory findings. CONCLUSION: The increased number of trophoblast cells in maternal peripheral blood and higher levels of sVCAM-1 correlate with the presence of PIH. The differences of sVCAM levels were significantly higher than those observed for sICAM. The results indicate an association between circulating trophoblasts and vascular endothelium activation, during PIH. PMID- 14748843 TI - Abnormal first trimester serum interleukin 18 levels are associated with a poor outcome in women with a history of recurrent miscarriage. AB - PROBLEM: How the maternal immune system adapts to tolerate the fetus is not fully understood, but a successful pregnancy is associated with the production of Th2 type cytokines and miscarriage is associated with the production of Th1-type cytokines. METHOD OF STUDY: Levels of interferon (IFN)-gamma, interleukin (IL)-4, IL-12 and IL-18 were measured in serum from 205 pregnant women of whom 115 pregnant women had a history of recurrent miscarriage. RESULTS: Compared with healthy pregnant women those who miscarried had increased serum levels of the Th1 associated cytokines IFN-gamma, IL-12 and IL-18. CONCLUSIONS: Increased levels of IL-18 appeared to be critical in early pregnancy and were able to discriminate between pregnancies that continued and those that end in miscarriage. PMID- 14748844 TI - Regulation of interleukin-6 fetoplacental levels could be involved in the protective effect of low-molecular weight heparin treatment on murine spontaneous abortion. AB - PROBLEM: CBA/J x DBA/2 abortion rate could be the consequence of a deficient local production of T helper (Th2) cytokines, which cause fetal wastage via fgl2 prothrombinase. Heparin reduces significantly the abortion rate in mice and recurrent spontaneous abortion (RSA) patients. We proposed to determine the effect of enoxaparin on the levels of local interleukin (IL)-6 during murine pregnancy. METHOD OF STUDY: Recombinant human IL-6 (rhIL-6) or enoxaparin were inoculated in CBA/J x DBA/2 pregnant mice on days 6.5-12.5. IL-6 levels in sera as well as in culture supernatants of day 9.5 fetoplacental units of CBA/J x BALB/c control mice or CBA/J x DBA/2 abortion combination were determined by enzyme-linked immunosorbent assay (ELISA) test. RESULTS: CBA/J x DBA/2 fetoplacental units secreted significantly lower levels of IL-6 with regard to CBA/J x BALB/c normal units. rhIL-6h and enoxaparin treatments decreased the resorption rate and regulated IL-6 fetoplacental levels. CONCLUSION: This study suggests that regulation of IL-6 fetoplacental levels could be involved in heparin-mediated anticoagulation protection against abortion. PMID- 14748845 TI - Serum concentrations of insulin-like growth factor-1, soluble tumor necrosis factor receptor-1 and angiogenin in endometriosis patients. AB - PROBLEM: Many soluble factors contributing to the pathophysiology of endometriosis are found at abnormal levels in patients suffering from the disease. We postulated that levels of these factors could also be altered in the serum of patients. We compared levels of insulin-like growth factor-1 (IGF-1), soluble form of tumor necrosis factor-alpha (TNF-alpha) receptor-1 (sTNFR-1) and angiogenin in the serum of patients with endometriosis and controls. METHOD OF STUDY: Levels of IGF-1, sTNFR-1 and angiogenin were measured by enzyme-linked immunosorbent assay in samples from 148 patients (77 cases and 71 controls) with diagnostic confirmed by laparoscopy. Correlations with demographic data and stage of the disease were evaluated and potential confounders in the study population were controlled. RESULTS: A significant increase in sTNFR-1 and angiogenin serum levels was observed in cases in comparison with controls, but only for patients in the follicular phase of the cycle. No significant difference was found in serum levels of IGF-1, sTNFR-1 and angiogenin between cases and controls in the luteal phase of the cycle. Correlations between levels of angiogenin and stage of the disease could also be observed. CONCLUSION: sTNFR-1 and angiogenin represent potential blood markers for endometriosis. PMID- 14748846 TI - The increased lysis of fetal cells in the mother after pregnancies complicated by pre-eclampsia or HELLP syndrome is not the result of a specific anti-fetal cytotoxicity of the mother. AB - PROBLEM: In pregnancies complicated by pre-eclampsia or hemolytisis elevated liver enzymes, low platelets (HELLP) syndrome, an increase of fetal DNA in the maternal serum indicates an increased lysis of fetal cells. Whether this cytolysis is the cause or the result of an increased specific cytotoxicity against the fetal cells is not yet known. METHODS: Ten mothers after healthy pregnancy, eight mothers after pregnancy complicated by prematurity (GA < 37 weeks), and eight mothers with pregnancies complicated by pre-eclampsia or HELLP syndrome and their male children were enrolled in the study. Fetally derived DNA in the maternal serum and the specific anti-fetal cytotoxicity of the maternal lymphocytes were measured. RESULTS: Detection of fetal DNA in maternal serum was significantly associated with pre-eclampsia/HELLP syndrome but not with anti fetal cytotoxicity of the maternal lymphocytes. CONCLUSION: The cytolysis of fetal cells in mothers after pregnancies complicated by pre-eclampsia/HELLP syndrome is neither the reason for nor the result of an increased specific anti fetal cytotoxicity. PMID- 14748847 TI - Good clinical practice is now obligatory in academic clinical drug research in the European Union. AB - By May 2004, all clinical trials in the European Union (EU) on medicinal products have to be initiated and conducted in compliance with the principles in the new directive on Good Clinical Practice (GCP). This requirement will also apply to non-commercial trials involving registered drugs and may therefore restrain the academic clinical drug research. In Denmark, three public GCP units connected in a national network and associated with the university hospitals in Copenhagen, Odense and Aarhus have been established. The GCP units offer academic researchers the necessary quality assurance and quality control systems to ensure that clinical drug research can be performed according to GCP. The Danish initiative is presented here as a contribution to the future work with implementation of the principles of GCP in academic clinical drug research in the European Union. PMID- 14748848 TI - Morphine-induced analgesia in the hot-plate test: comparison between NMRI(nu/nu) and NMRI mice. AB - NMRInu/nu mice are frequently used in cancer research, but their use in behavioural pain tests is unexplored. As behaviour of NMRI mice in pain tests is well-documented, a hot-plate test was performed comparing acute thermal nociception in NMRInu/nu and NMRI mice - untreated and morphine-treated - to estimate the usefulness of NMRInu/nu mice for further research on cancer pain. In both strains, morphine dose-dependently increased response latencies, number of animals reaching cut-off times and AUC values. Yet in NMRInu/nu mice, as compared to NMRI mice, all curves were shifted to the right. In order to be comparable, cut-off times must express a similar degree of baseline response augmentations. NMRInu/nu mice had substantially lower pre-drug latencies, indicating a lowered threshold for painful thermal stimuli, therefore effects of morphine in NMRInu/nu mice were also analysed using a lower cut-off time. Doing so, morphine resulted in similar effects in both strains. The effects were independent of hot-plate temperature, because similar results were obtained using temperatures of 50 and 55 degrees. The different morphine sensitivity of NMRInu/nu compared to NMRI mice primarily seems to depend upon differences in thermal threshold, probably induced by the different genotype of both strains. To determine whether cancer alters pain threshold or morphine analgesia, LoVo tumour-bearing NMRInu/nu mice were also tested. The tumour presence had no influence on withdrawal latencies or morphine efficacy. In general it can be concluded that NMRInu/nu mice with or without tumour can be used for nociceptive testing if baseline sensitivity is properly defined. PMID- 14748849 TI - Phencyclidine-induced behaviour in mice prevented by methylene blue. AB - Schizophrenia is a major public health problem that affects approximately 1% of the population worldwide. Schizophrenia-like symptoms can be induced in humans by phencyclidine (PCP), a drug with marked psychotomimetic properties. Phencyclidine disrupts prepulse inhibition of acoustic startle in rodents, a measure which has also been shown to be disrupted in schizophrenic patients. This effect is blocked by nitric oxide synthase (NOS) inhibitors, suggesting that nitric oxide plays an important role in this effect of phencyclidine. Methylene blue, a guanylate cyclase and nitric oxide syntase inhibitor, has shown therapeutic value as an adjuvant to conventional antipsychotics in the therapy of schizophrenia. The aim of the present study was to investigate if phencyclidine-(4 mg/kg)induced disruption of prepulse inhibition could be affected by methylene blue (50 or 100 mg/kg) in mice. Furthermore, the effect of methylene blue (50 mg/kg) on phencyclidine-(4 mg/kg)induced hyperlocomotion was investigated. The present study shows that phencyclidine readily disrupts prepulse inhibition in mice without affecting pulse-alone trials. It was also found that methylene blue prevents the decrease in prepulse inhibition caused by phencyclidine in a dose related manner. Furthermore, the increase in locomotor activity caused by phencyclidine was reduced by pretreatment with methylene blue. The results from the present study further support the suggestion that the nitric oxide synthase/guanylate cyclase pathway is involved in pharmacological and behavioural effects of phencyclidine. Since phencyclidine as well exerts psychotomimetic characteristics, agents that interfere with the nitric oxide synthase/guanylate cyclase pathway may be of therapeutic value also in the treatment of schizophrenia. PMID- 14748850 TI - Inhibition of voltage-sensitive sodium channels by the cannabinoid 1 receptor antagonist AM 251 in mammalian brain. AB - The cannabinoid 1 receptor antagonist AM 251 is known to block the inhibitory effects of endocannabinoids and synthetic cannabinoid agonists on transmitter release through an action at presynaptic cannabinoid 1 receptors in brain. We examined the ability of AM 251 to inhibit sodium channel-dependent functions and the binding of [3H]batrachotoxinin A 20-alpha-benzoate to sodium channels in mouse brain synaptic preparations. Depolarization of synaptoneurosomes by the sodium channel site 2-specific neurotoxin veratridine, which is abolished by tetrodotoxin, was found to be inhibited in a concentration-dependent fashion by AM 251 (IC50=8.9 microM). Veratridine-dependent (tetrodotoxin suppressible) release, of L-glutamic acid and GABA from synaptosomes was also reduced by AM 251 [IC50s=8.5 microM (L-glutamic acid), 9.2 microM (GABA)]. The binding of the radioligand [3H]batrachotoxinin A 20-alpha-benzoate to site 2 on sodium channels was displaced by AM 251 (IC50=11.2 microM). Scatchard analysis of binding showed that at its IC50, AM 251 increased (by 2.3 times) the KD of radioligand without altering Bmax, suggesting a competitive mechanism of inhibition by AM 251. Kinetic experiments indicated that AM 251 inhibits equilibrium binding by allosterically accelerating the dissociation of the [3H]-batrachotoxinin A 20 alpha-benzoate:sodium channel complex. Our data suggest that micromolar concentrations of AM 251 are capable of reducing neuronal excitability and inhibiting release of excitatory and inhibitory transmitters through blockade of voltage-sensitive sodium channels in brain. PMID- 14748851 TI - Anticonvulsant profile of flunarizine and relation to Na(+) channel blocking effects. AB - The present study will summarize our findings concerning the anticonvulsant properties of the Ca2+ channel blocker flunarizine in a variety of experimental models of epilepsy. Flunarizine exhibits anticonvulsant effects against tonic seizures induced by electroshock or various chemoconvulsants in mice, however, did not protect against pentylenetetrazol-induced clonic seizures. In the MES test, the efficacy of clinically established antiepileptics was increased by co medication. In the rotarod test, a minimal "neurotoxic" dose (TD50) of 18.0 mg/kg intraperitoneally was determined. In models of complex partial seizures like the hippocampal stimulation and the amygdala kindling in rats, flunarizine showed only a moderate activity. Thus, it can be suggested that the anticonvulsant potency of flunarizine in various screening tests is lower than that of standard antiepileptics such as carbamazepine and phenytoin. Concerning the possible mode of action, whole-cell patch-clamp experiments with cultured neonatal rat cardiomyocytes showed that flunarizine depressed the fast inward Na+ current in a concentration- and frequency-dependent manner well comparable with the action of phenytoin. It is concluded that the use-dependent inhibition of voltage-dependent Na+ channels may essentially contribute to the anticonvulsant activity of flunarizine in models for generalized tonic-clonic seizures. The clinical efficacy as add-on therapy is critically discussed in view of the present data. PMID- 14748852 TI - Gamma-hydroxybutyric acid, unlike gamma-aminobutyric acid, does not stimulate Gi/Go proteins in rat brain membranes. AB - gamma-Hydroxybutyric acid is a naturally occurring substance that may act as a neurotransmitter or neuromodulator to elicit several biological effects. Although the existence of a specific gamma-hydroxybutyric acid receptor has been postulated, the receptor protein itself has not been cloned yet. The current study was designed to elucidate whether gamma-hydroxybutyric acid receptors are functionally coupled with heterotrimeric G-proteins, especially Gi/Go family, by means of high-affinity GTPase activity and guanosine 5'-O-(3 [35S]thiotriphosphate) ([35S]GTPgammaS) binding assays in rat brain membranes. The stimulatory effects of GABAB receptor activation were always determined in parallel as a positive control. The selective GABAB receptor agonist (+/-) baclofen stimulated the high-affinity GTPase activity in cerebral cortical, hippocampal, and striatal membranes, whereas gamma-hydroxybutyric acid was inactive up to 1 mM in these brain regions. The optimum assay conditions for [35S]GTPgammaS binding to detect a receptor-mediated activation of G-proteins at the greatest signal to noise ratio were then probed as to the concentrations of constituents in the assay mixture (GDP, MgCl2, and NaCl) and incubation period. Even under such an optimized experimental condition, [35S]GTPgammaS binding was not altered by gamma-hydroxybutyric acid in the membranes prepared from cerebral cortex or hippocampus. On the other hand, the specific [35S]GTPgammaS binding was increased by GABAB receptor agonists in a concentration-dependent manner, which was competitively inhibited by CGP54626, a selective GABAB receptor antagonist. These results indicate that gamma-hydroxybutyric acid receptors, if any, are not associated with G-proteins, at least Gi/Go family. PMID- 14748853 TI - Effects of silymarin and vitamins E and C on liver damage induced by prolonged biliary obstruction in the rat. AB - Oxidative stress, in particular lipid peroxidation, induces collagen synthesis. Thus, we administered various antioxidants to bile duct-ligated rats for 28 days and lipid peroxidation, glutathione content, fibrosis, necrosis and cholestasis were evaluated. Extrahepatic cholestasis was induced by double ligation and section of the common bile duct. The study included eight groups (n=6), four groups were bile duct-ligated and received either vitamin C (50 mg/kg/day, orally), vitamin E (400 IU/rat/day, orally), silymarin (50 mg/kg/12 hr, orally) or vehicles; four groups were sham-operated controls. Collagen content was determined by measuring hydroxyproline in liver samples; malondialdehyde was used to estimate lipid peroxidation levels; reduced and oxidized glutathione were determined fluorometrically; alanine aminotransferase and bilirubins colorimetrically. Bilirubins increased several times, alanine aminotransferase once, reduced/oxidized glutathione ratio decreased three times, lipid peroxidation and collagen increased about three-times by biliary obstruction (p<0.05). Silymarin, vitamin E or C failed to prevent these effects significantly. It is not possible to clarify the role of oxidative stress in the fibrotic process induced by chronic biliary obstruction with the present results. Therefore, it seems reasonable to propose that a wide mixture of antioxidants, administered by the parenteral route (because cholestasis decreased the absorption of lipophilic compounds), is needed to counteract the oxidant stress produced by cholestasis. PMID- 14748854 TI - Do regulations limiting residents' work hours affect patient mortality? AB - OBJECTIVE: To conduct a statewide analysis of the effect of New York's regulations, limiting internal medicine and family practice residents' work hours, on patient mortality. DESIGN: Retrospective study of inpatient discharge files for 1988 (before the regulations) and 1991 (after the regulations). SETTING AND PATIENTS: Adult patients discharged from New York teaching hospitals (170214) and nonteaching hospitals (143,455) with a principal diagnosis of congestive heart failure, acute myocardial infarction, or pneumonia, for the years 1988 and 1991 (periods before and after Code 405 regulations went into law). Patients from nonteaching hospitals served as controls. MEASUREMENT: In-hospital mortality. RESULTS: Combined unadjusted mortality for congestive heart failure, acute myocardial infarction, and pneumonia patients declined between 1988 and 1991 in both teaching (14.1% to 13.0%; P =.0001) and nonteaching hospitals (14.0% to 12.5%; P =.0001). Adjusted mortality also declined between 1988 and 1991 in both teaching (odds ratio [OR], death 1991/1988, 0.868; 95% confidence interval [CI], 0.843 to 0.894; P =.0001) and nonteaching hospitals (OR, death 1991/1988, 0.853; 95% CI, 0.826 to 0.881; P =.0001). This beneficial trend toward lower mortality over time was nearly identical between teaching and nonteaching hospitals (P =.4348). CONCLUSION: New York's mandated limitations on residents' work hours do not appear to have positively or negatively affected in-hospital mortality from congestive heart failure, acute myocardial infarction, or pneumonia in teaching hospitals. PMID- 14748855 TI - Hospitalists as teachers. AB - OBJECTIVE: To compare evaluations of teaching effectiveness among hospitalist, general medicine, and subspecialist attendings on general medicine wards. DESIGN: Cross-sectional. SETTING: A large, inner-city, public teaching hospital. PARTICIPANTS: A total of 423 medical students and house staff evaluating 63 attending physicians. MEASUREMENTS AND MAIN RESULTS: We measured teaching effectiveness with the McGill Clinical Tutor Evaluation (CTE), a validated 25 item survey, and reviewed additional written comments. The response rate was 81%. On a 150-point composite measure, hospitalists' mean score (134.5 [95% confidence interval (CI), 130.2 to 138.8]) exceeded that of subspecialists (126.3 [95% CI, 120.4 to 132.1]), P =.03. General medicine attendings (135.0 [95% CI, 131.2 to 138.8]) were also rated higher than subspecialists, P =.01. Physicians who graduated from medical school in the 1990s received higher scores (136.0 [95% CI, 133.0 to 139.1]) than did more distant graduates (129.1 [95% CI, 125.1 to 133.1]), P =.006. These trends persisted after adjusting for covariates, but only year of graduation remained statistically significant, P =.05. Qualitative analysis of written remarks revealed that trainees valued faculty who were enthusiastic teachers, practiced evidence-based medicine, were involved in patient care, and developed a good rapport with patients and other team members. These characteristics were most often noted for hospitalist and general medicine attendings. CONCLUSIONS: On general medicine wards, medical students and residents considered hospitalists and general medicine attendings to be more effective teachers than subspecialists. This effect may be related to the preferred faculty members exhibiting specific characteristics and behaviors highly valued by trainees, such as enthusiasm for teaching and use of evidence based medicine. PMID- 14748856 TI - Adherence counseling practices of generalist and specialist physicians caring for people living with HIV/AIDS in North Carolina. AB - CONTEXT: National guidelines recommend that practitioners assess and reinforce patient adherence when prescribing antiretroviral (ART) medications, but the extent to which physicians do this routinely is unknown. OBJECTIVE: To assess the adherence counseling practices of physicians caring for patients with HIV/AIDS in North Carolina and to determine characteristics associated with providing routine adherence counseling. DESIGN: A statewide self-administered survey. SETTING AND PARTICIPANTS: All physicians in North Carolina who prescribed a protease inhibitor (PI) during 1999. Among the 589 surveys sent, 369 were returned for a response rate of 63%. The 190 respondents who reported prescribing a PI in the last year comprised the study sample. MAIN OUTCOME MEASURES: Physicians reported how often they carried out each of 16 adherence counseling behaviors as well as demographics, practice characteristics, and attitudes. RESULTS: On average, physicians reported spending 13 minutes counseling patients when starting a new 3 drug ART regimen. The vast majority performed basic but not more extensive adherence counseling; half reported carrying out 7 or fewer of 16 adherence counseling behaviors "most" or "all of the time." Physicians who reported conducting more adherence counseling were more likely to be infectious disease specialists, care for more HIV-positive patients, have more time allocated for an HIV visit, and to perceive that they had enough time, reimbursement, skill, and office space to counsel. After also controlling for the amount of reimbursement and availability of space for counseling, physicians who were significantly more likely to perform a greater number of adherence counseling practices were those who 1). cared for a greater number of HIV/AIDS patients; 2). had more time allocated for an HIV physical; 3). felt more adequately skilled; and 4). had more positive attitudes toward ART. CONCLUSIONS: This first investigation of adherence counseling practices in HIV/AIDS suggests that physicians caring for patients with HIV/AIDS need more training and time allocated to provide antiretroviral adherence counseling services. PMID- 14748857 TI - Health care resource utilization associated with a diabetes center and a general medicine clinic. AB - OBJECTIVE: Studies have proposed that the features of diabetes clinics may decrease hospital utilization and costs by reducing complications and providing more efficient outpatient care. We compared the health care utilization associated with a diabetes center (DC) and a general medicine clinic (GMC). DESIGN: Retrospective cohort study. SETTING: An urban academic medical center. PATIENTS/PARTICIPANTS: Type 2 diabetes patients (N = 601) under care in a DC and GMC before March 1996. MEASUREMENTS AND MAIN RESULTS: We compared baseline patient characteristics and outpatient care for the period of March 1996 to August 1997. Using administrative data from March 1996 to October 2000, we compared the probability of a hospitalization, length of stay, costs of hospitalizations, the probability of an emergency room visit, and costs of emergency room visits. Diabetes center patients had a longer mean duration of diabetes (12 years vs 6 years, P <.01), more baseline microvascular disease (65% vs 44%, P <.01), and higher baseline glucose levels (hemoglobin A1c 8.6% vs 7.9%, P <.01) than GMC patients. Diabetes center patients received more intensive outpatient care directed toward glucose monitoring and control. In all crude and adjusted analyses of hospitalizations and emergency room visits, we found no statistically significant differences for inpatient utilization or cost outcomes comparing clinic populations. CONCLUSIONS: Diabetes center attendance did not have a definitive positive or negative impact on inpatient resource utilization over a 4-year period. However, DC patients had more severe diabetes but no greater hospital utilization compared with GMC patients. Clear demonstration of the clinical and financial benefits of features of diabetes centers will require long-term controlled trials of interventions that promote comprehensive diabetes care, including cardiovascular prevention. PMID- 14748858 TI - Resource use in treating alcohol- and drug-related diagnoses. AB - OBJECTIVE: This study examined the variations in hospital resource use in the treatment of alcohol and drug diagnoses. Specifically, the study tested 2 hypotheses: 1). patients treated in teaching hospitals will have shorter lengths of stay and lower hospital charges than patients treated in nonteaching hospitals; and 2). patients treated in hospitals with more experience treating these conditions will have shorter lengths of stay and lower hospital charges. DESIGN: A retrospective cross-sectional study design used data from the 1996 Health Care Utilization Project to test the proposed hypotheses. PATIENTS/PARTICIPANTS: The population for this study consisted of patients over 18 years old with an acute alcohol- or drug-related discharge diagnostic related group code. MEASUREMENT AND MAIN RESULTS: The variables of interest were teaching hospital status, as defined by the Council of Teaching Hospitals, and hospital experience, defined as the ratio of alcohol- and drug-related diagnoses to the hospital's total admissions. Measures of hospital resource use included the patient's length of stay and total hospital charges. Patients treated at hospitals with relatively more experience in treating alcohol- and drug-related diagnoses had 10.3% (US dollars 321) lower total charges (P =.017). CONCLUSIONS: Similar to research for high-volume surgical hospitals, these findings confirm that hospitals that have greater experience with complex medical conditions such as alcohol and drug intoxication and withdrawal may be more efficient. This important finding provides a rationale for further exploration of the key factors associated with higher quality and more efficient care for complex medical conditions. PMID- 14748859 TI - Use of mind-body medical therapies. AB - OBJECT: Research demonstrating connections between the mind and body has increased interest in the potential of mind-body therapies. Our aim was to examine the use of mind-body therapies, using data available from a national survey. DESIGN: Analysis of a large nationally representative dataset that comprehensively evaluated the use of mind-body therapies in the last year. SETTING: United States households. PATIENTS/PARTICIPANTS: A total of 2055 American adults in 1997-1998. INTERVENTIONS: Random national telephone survey. MEASURES AND MAIN RESULTS: We obtained a 60% weighted overall response rate among eligible respondents. We found that 18.9% of adults had used at least 1 mind-body therapy in the last year, with 20.5% of these therapies involving visits to a mind-body professional. Meditation, imagery, and yoga were the most commonly used techniques. Factors independently and positively associated with the use of mind body therapies in the last year were being 40 to 49 years old (adjusted odds ratio [AOR], 2.03; 95% confidence interval [CI], 1.33 to 3.10), being not married (AOR, 1.78; 95% CI, 1.34 to 2.36), having an educational level of college or greater (AOR, 2.21; 95% CI, 1.57 to 3.09), having used self-prayer for a medical concern (AOR, 2.53; 95% CI, 1.87 to 3.42), and having used another complementary medicine therapy in the last year (AOR, 3.77; 95% CI, 2.74 to 5.20). While used for the full array of medical conditions, they were used infrequently for chronic pain (used by 20% of those with chronic pain) and insomnia (used by 13% of those with insomnia), conditions for which consensus panels have concluded that mind body therapies are effective. They were also used by less than 20% of those with heart disease, headaches, back or neck pain, and cancer, conditions for which there is strong research support. Mind-body therapies were generally used concomitantly with conventional care: 90% of those using a mind-body therapy in the last year had seen a physician and 80% of mind-body therapies used were discussed with a physician. CONCLUSIONS: Although mind-body therapies were commonly used, much opportunity exists to increase use of mind-body therapies for indications with demonstrated efficacy. PMID- 14748860 TI - Relationship between conflicts of interest and research results. AB - CONTEXT: To date, research regarding the influence of conflicts of interest on the presentation of findings by researchers has been limited. OBJECTIVE: To evaluate the sources of funding for published manuscripts, and association between reported findings and conflicts of interest. METHODS: Data from both print and electronic issues of The New England Journal of Medicine (NEJM) and The Journal of the American Medical Association (JAMA) were analyzed for sources of funding, areas of investigation, conflict of interest (COI), and presentation of results. We reviewed all original manuscripts published during the year 2001 within NEJM (N = 193) and JAMA (N = 205). We use 3 definitions for COI in this paper: a broadly defined criterion, the criterion used by The International Council of Medical Journal Editors (ICMJE), and a criterion defined by the authors. RESULTS: Depending on the COI criteria used, 16.6% to 32.6% of manuscripts had 1 or more author with COI. Based on ICMJE criterion, 38.7% of studies investigating drug treatments had authors with COI. We observed a strong association between those studies whose authors had COI and reported positive findings (P <.001). When controlling for sample size, study design, and country of primary authors, we observed a strong association between positive results and COI (ICMJE definition) among all treatment studies (adjusted odds ratio [OR], 2.35; 95% confidence interval [CI], 1.08 to 5.09) and drug studies alone (OR, 2.64; 95% CI, 1.09 to 6.39). CONCLUSION: COI is widespread among the authors of published manuscripts and these authors are more likely to present positive findings. PMID- 14748861 TI - Development and testing of the health care system distrust scale. AB - BACKGROUND: Distrust of the health care system may be a significant barrier to seeking medical care, adhering to preventive health care and treatment regimens, and participating in medical research. OBJECTIVE: To describe the development and psychometric testing of an instrument (the Health Care System Distrust Scale) to measure distrust of the health care system. METHODS: Scale development involved 2 phases. In Phase 1, a pilot instrument was developed based on a conceptual model of health care-related distrust. Draft items were created using focus group sessions with members of the general public, literature review, and expert opinion. Draft items were pilot tested with 55 individuals waiting to be assigned to jury duty at the Municipal Court of Philadelphia. A priori, candidate items for elimination or revision included those with >5% missing data, extremely low or high interitem or item-total correlations, or those having a negative effect on the scale's internal consistency. In Phase 2, we conducted a survey of 400 prospective jurors to assess the reliability and validity of the final scale scores. RESULTS: In Phase 1, a 10-item scale was constructed that included 4 items measuring honesty, 2 items measuring confidentiality, 2 items measuring competence, and 2 items measuring fidelity. The participants in Phase 2 had a mean age of 41 years. Forty-three percent were African-American, 45% white, and 4% Hispanic. Scores on the Health Care System Distrust scale ranged from 12 to 46 with a possible range from 10 to 50. The mean score was 29.4 with a standard deviation of 6.33. No item had over 5% missing data. Internal consistency (Cronbach's alpha) was 0.75. Item-total correlations ranged from 0.27 to 0.57. Principal components analysis revealed 1 general component accounting for 32% of the variance. Nine of the variables had loadings higher than 0.40. As predicted, distrust of the health care system was higher among African Americans than whites and was inversely correlated with trust in personal physicians. CONCLUSIONS: Initial testing suggests that we developed an instrument with valid and reliable scores in order to measure distrust of the health care system. Future research is needed to evaluate the validity and reliability of the Health Care System Distrust scale among diverse populations. This instrument can facilitate the investigation of the prevalence, causes, and effects of health care system distrust in the United States. PMID- 14748862 TI - Facilitating scholarly writing in academic medicine. AB - Scholarly writing is a critical skill for faculty in academic medicine; however, few faculty receive instruction in the process. We describe the experience of 18 assistant professors who participated in a writing and faculty development program which consisted of 7 monthly 75-minute sessions embedded in a Collaborative Mentoring Program (CMP). Participants identified barriers to writing, developed personal writing strategies, had time to write, and completed monthly writing contracts. Participants provided written responses to open-ended questions about the learning experience, and at the end of the program, participants identified manuscripts submitted for publication, and completed an audiotaped interview. Analysis of qualitative data using data reduction, data display, and conclusion drawing/verification showed that this writing program facilitated the knowledge, skills, and support needed to foster writing productivity. All participants completed at least 1 scholarly manuscript by the end of the CMP. The impact on participants' future academic productivity requires long-term follow-up. PMID- 14748863 TI - The future of general internal medicine. Report and recommendations from the Society of General Internal Medicine (SGIM) Task Force on the Domain of General Internal Medicine. AB - The Society of General Internal Medicine asked a task force to redefine the domain of general internal medicine. The task force believes that the chaos and dysfunction that characterize today's medical care, and the challenges facing general internal medicine, should spur innovation. These are our recommendations: while remaining true to its core values and competencies, general internal medicine should stay both broad and deep-ranging from uncomplicated primary care to continuous care of patients with multiple, complex, chronic diseases. Postgraduate and continuing education should develop mastery. Wherever they practice, general internists should be able to lead teams and be responsible for the care their teams give, embrace changes in information systems, and aim to provide most of the care their patients require. Current financing of physician services, especially fee-for-service, must be changed to recognize the value of services performed outside the traditional face-to-face visit and give practitioners incentives to improve quality and efficiency, and provide comprehensive, ongoing care. General internal medicine residency training should be reformed to provide both broad and deep medical knowledge, as well as mastery of informatics, management, and team leadership. General internal medicine residents should have options to tailor their final 1 to 2 years to fit their practice goals, often earning a certificate of added qualification (CAQ) in special generalist fields. Research will expand to include practice and operations management, developing more effective shared decision making and transparent medical records, and promoting the close personal connection that both doctors and patients want. We believe these changes constitute a paradigm shift that can benefit patients and the public and reenergize general internal medicine. PMID- 14748864 TI - Effectiveness of screening and treatment for depression in ambulatory indigent patients. AB - OBJECTIVE: To determine the effectiveness of screening and treatment for depression among ambulatory indigent patients visiting resident physicians. DESIGN: Two-group randomized trial (N = 33 intervention, N = 28 usual care) with baseline, 6-month, and 12-month outcome measurements. SETTING: Internal Medicine Residency Clinic. PATIENTS: Clinic patients over 18 years of age who screened positive for depression on the PRIME-MD during a visit to their resident physician. Patients were not receiving treatment nor seeking care for any emotional problems. All patients were either enrolled in Medicaid or had income below the poverty line. INTERVENTION: Resident physicians were educated to follow AHCPR (AHRQ; Agency for Healthcare Research and Quality) guidelines for diagnosis and treatment of depression in a primary care setting. For the intervention group patients, a screening nurse advised residents regarding the positive screen, handed them a standardized protocol outline, and attempted to arrange behavioral care. The patients in the usual care group were provided the results of the screen by the screening nurse before their visit with the resident, and advised to seek care for their symptoms. MAIN RESULTS: Results for the primary outcome of depression symptoms measured with the Beck Depression Inventory (BDI) demonstrated that intervention was successful in reducing symptoms relative to usual care (difference = -4.9 BDI points, P =.05, 95% confidence interval [CI], 9.8 to -0.005 effect size = -0.41). During the 12-month follow-up, 70% of intervention patients were treated for depression (of these, 91% with antidepressants), while 15% of usual care patients were treated with antidepressants for depression. Another 18% of the usual care group had depression noted, but no treatment was identified. BDI differences between intervention and control groups were similar at the 6- and 12-month measures. Quality of life and costs were also measured, but differences between the groups were not significant in this regard. CONCLUSION: Screening and treatment for depression by resident physicians was successful in reducing symptoms relative to usual care in an indigent population. Almost twice as many intervention patients as usual care controls demonstrated a substantial reduction (10 BDI points) in symptoms related to depression. PMID- 14748865 TI - Reasons physicians accepted or declined smallpox vaccine, February through April, 2003. AB - From February to April 2003, we performed an e-mail-based survey to assess responses of physicians at Yale University to being offered smallpox vaccine. Of 58 respondents, 3 (5%) had been or intended to be vaccinated. Reasons cited for declining vaccination included: belief that benefits did not outweigh risks (55%), belief that the vaccination program was unnecessary (18%), desire to wait and see what side effects occurred in vaccinees (11%), and worries about compensation or liability (7%). Most (94%) considered risks to themselves, family, or patients in their decision. Only 3% thought a smallpox attack in the next 5 years was likely or very likely. Physicians did not accept the smallpox vaccine because they did not believe the potential benefits were sufficient. PMID- 14748866 TI - Luxury primary care, academic medical centers, and the erosion of science and professional ethics. AB - Medical schools and teaching hospitals have been hit particularly hard by the financial crisis affecting health care in the United States. To compete financially, many academic medical centers have recruited wealthy foreign patients and established luxury primary care clinics. At these clinics, patients are offered tests supported by little evidence of their clinical and/or cost effectiveness, which erodes the scientific underpinnings of medical practice. Given widespread disparities in health, wealth, and access to care, as well as growing cynicism and dissatisfaction with medicine among trainees, the promotion by these institutions of an overt, two-tiered system of care, which exacerbates inequities and injustice, erodes professional ethics. Academic medical centers should divert their intellectual and financial resources away from luxury primary care and toward more equitable and just programs designed to promote individual, community, and global health. The public and its legislators should, in turn, provide adequate funds to enable this. Ways for academic medicine to facilitate this largesse are discussed. PMID- 14748867 TI - Challenges and opportunities for general internal medicine. PMID- 14748868 TI - In pursuit of optimal duty hours and resident experiences. PMID- 14748869 TI - Role of endoscopic ultrasonography in diseases of the extrahepatic biliary system. AB - Endoscopic ultrasonography (EUS) is one of the most recent advances in gastrointestinal endoscopy. Available EUS devices include echoendoscopes, such as radial scanning and linear array echoendoscopes, and catheter ultrasound probes. Endoscopic ultrasonography has various applications, such as staging of gastrointestinal malignancy, evaluation of submucosal tumors, and has grown to be an important modality in evaluating the pancreaticobiliary system. With regard to the biliary system, EUS is useful for the detection and staging of ampullary tumors, detection of microlithiasis and choledocholithiasis and evaluation of benign and malignant bile-duct strictures. Endoscopic ultrasonography may be used as an adjunct to transabdominal ultrasound for the detection and characterization of gallbladder polyps. In addition, EUS is helpful in the staging of gallbladder cancer as well as in diagnosing anomalous pancreaticobiliary junction with its associated pancreatobiliary diseases. The present paper reviews the current applications of EUS for imaging and intervention in diseases of the extrahepatic biliary system. PMID- 14748870 TI - Effect of Helicobacter pylori eradication on gastroesophageal function. AB - BACKGROUND: To elucidate the cause of possible occurrence of reflux esophagitis after Helicobacter pylori eradication, gastric and esophageal function among H. pylori infected Japanese patients were evaluated both before and after eradication therapy. METHODS: Nine H. pylori-positive patients were studied before and 6 months after successful H. pylori eradication. Studies included gastric emptying, esophageal manometry, gastric and esophageal pH monitoring as well as measuring serum levels of gastrin, pepsinogen I and pepsinogen II. RESULTS: Helicobacter pylori eradication was associated with a significant change in serum gastrin and pepsinogen levels, consistent with the improvement in mucosal inflammation. There was no significant change in gastric emptying, fasting or postprandial lower esophageal sphincter (LES) pressure, esophageal primary peristaltic contractions, frequency of transient LES relaxation, or gastroesophageal reflux, as assessed by 24 h pH monitoring. The percent time of the gastric pH>4 at night decreased significantly. A 41-year-old male developed erosive gastroesophageal reflux disease (GERD) (Los Angeles Classification Grade A) after eradication. Physiological studies showed he had abnormal esophageal motility prior to H. pylori eradication. CONCLUSIONS: With the exception of gastric pH at night, most patients did not experience a significant change in gastric or esophageal function after H. pylori eradication. Development of GERD post H. pylori eradication likely reflects an increase in the acidity of the refluxate superimposed on pre-existing abnormalities in gastroesophageal motility. PMID- 14748871 TI - Gastroesophageal reflux disease related to diabetes: Analysis of 241 cases with type 2 diabetes mellitus. AB - BACKGROUND AND AIM: We examined the incidence of symptomatic gastroesophageal reflux disease (GERD) in patients with type 2 diabetes mellitus (DM). METHODS: Patients comprised those with DM or chronic hepatitis C (CHC) who visited Osaka Prefectural General Hospital in the same study period. Reflux symptoms were examined using a self-administered questionnaire. A total score of 4 or more was considered an indication of symptomatic GERD. Disease duration, hemoglobinA1c and diabetic complications were assessed. RESULTS: Patients with DM (n=241) or CHC (n=42) were recruited for the study. Of the 241 patients with DM, 100 (41.5%) reported experiencing upper gastrointestinal symptoms, whereas only 9 of 42 (21.4%) patients with CHC reported upper gastrointestinal symptoms (P=0.0137). Sixty-one patients (25.3%) with DM had reflux symptoms but only four patients (9.5%) with CHC reported reflux symptoms. The incidence of symptomatic GERD was significantly higher in patients with DM than in those with CHC (P=0.0219). Patients with DM for less than 5 years had a 2.4-fold higher incidence of GERD than patients with CHC. The incidence tended to rise with increased disease duration. Patients with diabetic complications reported reflux symptoms more frequently. The incidence decreased, however, in DM patients who had these conditions for more than 16 years. CONCLUSIONS: Type 2 diabetes mellitus is a risk factor for symptomatic GERD. In DM patients, use of oral hypoglycemic agents, body mass index, disease duration and the quality of diabetic control influenced the incidence of GERD. PMID- 14748872 TI - Prevalence of colonic polyps is not increased in patients with acromegaly: analysis of 60 patients from India. AB - BACKGROUND: There are conflicting data on the prevalence of colorectal adenomas in patients with acromegaly. It has been suggested that the risk of colorectal adenomas may be dependent on the geographic and ethnic origin of the patients. MATERIALS: Sixty consecutive patients with active acromegaly due to somatotropinoma underwent colonoscopy prior to definitive surgery. They included 35 men and 25 women with a mean (+/- SD) age of 37.4 +/- 13.2 years and a lag time between symptomatology and diagnosis of acromegaly of 64.6 +/- 58.0 months. The control group included 160 patients (88 men, 72 women; mean age, 38.2 +/- 14.0 years) with a diagnosis of irritable bowel syndrome who were also subjected to colonoscopy. None of these patients had previous or family history of colonic neoplasm, bleeding or colonic surgery. RESULTS: Colonoscopic examination was complete to the cecum in 52 patients (88%), and to the splenic flexure in the remaining 12%. In the control group, it was complete to the cecum in 144 patients (90%). Four of the acromegalic patients (6.7%) and five in the control group (3.1%; P=0.24) had hyperplastic polyps. No patient in either of the groups had adenomatous polyps or colonic adenocarcinoma. The group of acromegalic patients with and without polyps did not differ significantly in age, duration of disease, growth hormone levels or glycemic status. The number of skin tags, however, was significantly higher (P=0.04) in the polyp group as compared to those without polyps. CONCLUSION: The present study has failed to demonstrate the high prevalence rate of colonic neoplasia in patients with acromegaly as compared to reports from the Western world. PMID- 14748873 TI - Intestinal inflammation alters the susceptibility to pentylenetetrazole-induced seizure in mice. AB - BACKGROUND: Inflammatory bowel disorders are associated with increased incidence of seizures. Alteration in the endogenous opioid system and overproduction of nitric oxide have been implicated in colitis. The possible contribution of opioid receptors and nitric oxide to increased seizure susceptibility was examined in a putative model of intestinal inflammation. METHODS: The alterations in clonic seizure threshold, induced by pentylenetetrazole, following the induction of intestinal inflammation by the administration of two consecutive daily oral doses of croton oil, was evaluated in mice. This model was used to evaluate the effects of pretreatment with opioid receptor antagonist naltrexone (10 mg/kg, once daily for 4 days or a single dose on the test day), non-specific nitric oxide synthase inhibitor NG-nitro-L-arginine methyl ester (10 mg/kg, once daily), and specific inducible nitric oxide synthase inhibitor aminoguanidine (100 mg/kg, once daily) on seizure threshold in intestinal inflammation. RESULTS: A significant decrease in clonic seizure threshold was observed in mice with intestinal inflammation compared to the control group. Chronic pretreatment with naltrexone per se did not cause any significant change in seizure threshold, but it significantly restored the threshold in croton oil-treated mice to that of the control animals. However, acute naltrexone pretreatment (on the test day) could not restore the decreased seizure threshold to control level. Chronic pretreatment with neither NG-nitro-L-arginine methyl ester nor aminoguanidine altered the seizure threshold in the control animals and in mice treated with croton oil. CONCLUSIONS: Experimental croton oil-induced intestinal inflammation leads to a proconvulsant effect, which may have clinical relevance. Chronic alterations mediated by endogenous opioids may be involved in this process, though a direct opioid receptor-mediated effect is unlikely. Nitric oxide synthesis, via constitutive or inducible pathways, is not involved in this increased susceptibility. PMID- 14748874 TI - Can somatostatin prevent post-ERCP pancreatitis? Results of a randomized controlled trial. AB - BACKGROUND: Acute pancreatitis is the most common complication of endoscopic retrograde cholangiopancreatography (ERCP), occurring in 1-10% of patients. Several substances have been used, with negative results, in an attempt to prevent this complication. METHODS: We performed a double-blind randomized trial in 372 consecutive patients undergoing diagnostic or therapeutic ERCP to evaluate the role of somatostatin in preventing post-ERCP pancreatitis. The first group received continuous somatostatin infusion for 12 h starting 30 min before ERCP, the second group received a bolus intravenous injection of somatostatin at the time of cannulation of the papilla, and the third group received a placebo. RESULTS: Two patients in each of the somatostatin groups (1.7%) and 12 patients in the placebo group (9.8%) developed pancreatitis (P<0.05). Serum amylase levels 5 and 24 h after the procedure were lower in both groups that received somatostatin than in the placebo group (P<0.05). CONCLUSION: Somatostatin is useful in preventing post-ERCP pancreatitis. Further studies must be designed to investigate the cost-effectiveness of the drug and to determine the ideal administration route and dosage. PMID- 14748875 TI - Surveillance for newly acquired hepatitis C in Australia. AB - BACKGROUND: The purpose of the present paper was to determine recent patterns of hepatitis C virus (HCV) transmission in Australia through a national system of enhanced surveillance of newly acquired hepatitis C. METHODS: Demographic, clinical, and risk behavior information on newly acquired hepatitis C cases from 1997 to 2000 was collected. Newly acquired hepatitis C included cases of HCV antibody sero-conversion within a 12 month period and acute clinical hepatitis C cases. RESULTS: Nine hundred and twelve cases of newly acquired hepatitis C were identified, representing 2.8% of all HCV notifications for this period. The majority of cases (72%) were diagnosed in people aged between 20 and 39 years. Injecting drug use was reported in the vast majority of cases (93%), with sexual transmission (2%) and tattooing (2%) reported in small numbers. HCV antibody sero conversion was the mode of diagnosis in most cases (78%). CONCLUSIONS: Injecting drug use is the main route of HCV transmission in Australia. As only a small proportion of HCV infections are detected as newly acquired, enhanced surveillance procedures, including increased regular HCV testing of at-risk populations are required to more effectively monitor recent patterns of transmission. PMID- 14748876 TI - High frequency of chronic end-stage liver disease and hepatocellular carcinoma in a Hispanic population. AB - BACKGROUND: Texas has the highest mortality from hepatocellular carcinoma in the USA. Because end-stage liver disease mortality is most marked in the Hispanic population in the Lower Rio Grande Valley, the aim of the present paper was to estimate the prevalence of end-stage liver disease and associated factors in Brownsville. METHODS: A cross-sectional study was carried out, abstracting medical charts in a community-based clinic and a hospital. A matched case-control analysis was performed. Cases had an International Classification of Disease (9th revision; ICD9) code 155.0 or 571.0-9 (primary liver cancer or chronic liver disease) recorded; age- and sex-matched controls did not have these codes. A total of 176 cases and 352 controls was collected. The main outcome measure was the prevalence and risk factors for end-stage liver disease. RESULTS: Conservative prevalence of end-stage liver disease: 126/100,000 (386/100,000, male). Eleven out of 176 had hepatocellular carcinoma. Median age was 57 years; 72% male, 94% Hispanic. Among significant risk factors were history of hepatitis (odds ratio (OR): 19.3; 95% confidence interval (CI): 8.0-46.4), any history of alcohol use (OR: 6.6; 95%CI: 4-10.8) and history of illegal drug use (OR: 1.9; 95%CI: 1.2-2.9). Fifteen cases with no known risk factors were classified as cryptogenic cirrhosis. Only four cases out of 176 had been referred for liver transplant. CONCLUSIONS: The prevalence of end-stage liver disease in the Lower Rio Grande Valley is extremely high, with no single satisfactory explanation and with acute health disparities. Careful follow up of cryptogenic cirrhosis in this population may or may not lead to a new source of liver disease. PMID- 14748877 TI - Cartilage oligomeric matrix protein expression in hepatocellular carcinoma and the cirrhotic liver. AB - BACKGROUND: Cartilage oligomeric matrix protein (COMP) is the fifth member of the thrombospondin family of extracellular, calcium-binding proteins. It was initially isolated and characterized in cartilage tissue, where it is thought to contribute to the extracellular matrix composition and cell-extracellular matrix interaction. In the present study the expression of COMP was investigated in normal liver (n=19), liver cirrhosis (n=14) and hepatocellular carcinoma (HCC; n=16) tissues, both at the mRNA and protein level. METHODS AND RESULTS: By northern blot and western blot analysis, COMP was absent or rarely expressed in the normal liver and liver cirrhosis tissues, but significantly overexpressed in HCC tissue samples. The COMP mRNA overexpression in HCC was not related to the clinical stage or tumor grade. By in situ hybridization and immunohistochemistry analysis, COMP mRNA and protein expression were localized within the cytoplasm of the tumor cells. CONCLUSION: COMP is highly expressed within the tumor cells of HCC, suggesting that COMP might play a role in the pathophysiology of this disease. PMID- 14748878 TI - Ingestion of resistant starch protects endotoxin influx from the intestinal tract and reduces D-galactosamine-induced liver injury in rats. AB - BACKGROUND AND AIMS: The aim of the present study was to examine the protective effect of a dietary high-amylose cornstarch (HAS) against D-galactosamine (D GalN)-induced liver injury, focusing specifically on intestinal endotoxin translocation. METHODS: Male Wistar rats fed a HAS-free basal diet or a 30% HAS supplemented diet were injected intraperitoneally with D-GalN. Serum transaminase activities, serum concentrations of tumor necrosis factor (TNF)-alpha, and portal venous endotoxin concentrations were determined at various time points. Ileal mucosal proliferation, small intestinal immunoglobulin (Ig)A and mucin, and the size of the cecal short-chain fatty acids (SCFA) pool were also determined. RESULTS: High-amylose cornstarch ingestion significantly reduced the increase in serum transaminase activities at 22 h after the injection of D-GalN. Rats fed the HAS diet showed a greater cecal SCFA production as measured by pool size than those fed the basal diet. Luminal IgA and mucin content were significantly greater in rats fed the HAS diet. Protein, DNA and RNA contents in the ileal mucosa were also higher in rats fed the 30% HAS diet. In a further experiment, portal venous endotoxin concentrations in rats fed the basal diet reached 72 ng/L at 4 h after D-GalN administration, but endotoxin was not detected in rats fed the HAS diet. At this time, portal endotoxin concentrations were significantly and positively correlated with the serum concentrations of TNF-alpha and serum alanine aminotransferase activities. CONCLUSION: These data support the view that HAS ingestion may reduce D-GalN-induced liver injury as a result of an inhibitory effect on endotoxin influx from the intestinal tract, at least in part as a result of alterations in the mucosal barrier functions. PMID- 14748879 TI - Role of gamma-glutamyl transferase activity in patients with chronic hepatitis C virus infection. AB - BACKGROUND: Increased serum gamma-glutamyl transferase (GGT) levels are frequently observed in chronic hepatitis C virus (HCV) infection. However, the significance of this finding remains unclear. The purpose of the present paper was to assess the relationship between GGT levels and clinical, biochemical and histological features in chronic HCV-infected carriers. METHODS: Patients with a liver biopsy presenting anti-HCV and HCV-RNA were evaluated. Age, gender, risk factors of transmission, serum alanine aminotransferase (ALT), GGT and alkaline phosphatase (ALP) levels and histological features were assessed in all. Data were analyzed statistically by the chi2 test and multivariate logistic regression analysis. RESULTS: Among 201 patients studied, elevated GGT levels and bile duct damage were observed in 48% and 35% of them, respectively. No association was seen between GGT level and bile duct damage or between GGT level and hepatic steatosis. Initially, age >40 years (P=0.007), elevated ALT (P=0.01), grading of inflammatory activity (P=0.004) and staging of fibrosis (P<0.001) were found to be associated with elevated GGT levels. After multivariate regression analysis, histology grading 3 and 4 inflammation activity (P=0.01) and staging 3 and 4 fibrosis (P=0.01) remained independently associated with elevated GGT level. CONCLUSIONS: A significant number of patients with chronic HCV infection had elevated serum GGT levels. Furthermore, this enzyme seemed to be useful as an indirect marker of more advanced liver disease in chronic hepatitis C. PMID- 14748880 TI - Immunoactivative role of indoleamine 2,3-dioxygenase in human hepatocellular carcinoma. AB - BACKGROUND: Indoleamine 2,3-dioxygenase (IDO) is a tryptophan catabolic enzyme. Recent studies have focused on the immunoregulatory role of IDO in mononuclear cells. The role of IDO in hepatocellular carcinoma (HCC) cell lines and HCC patients was examined. METHODS: The expression of IDO mRNA in peripheral blood mononuclear cells (PBMC) cocultured with HCC cell lines was detected by reverse transcriptase-polymerase chain reaction (RT-PCR). The cytotoxicity of PBMC against HCC cell lines cultured with and without IDO inhibitor was examined by sodium 51chromate release assay. In the tumor portion of 21 HCC patients, the expression of mRNA of IDO, tryptophan 2,3-dioxygenase and some cytokines was detected by RT-PCR. The expression and distribution of IDO protein in HCC specimens was analyzed by immunohistochemistry. RESULTS: The IDO mRNA was strongly induced in PBMC cocultured with HepG2 and PLC/PRF/5 and faintly induced in PBMC cocultured with Hep3B and HuH7. The cytotoxicity of PBMC against HCC cell lines was directly proportional to the level of expression of IDO mRNA and reduced by IDO inhibitor. The expression of IDO mRNA in the tumor portion was detected in 12 out of 21 HCC patients. Immunohistochemistry revealed that the IDO positive cells were identified to be tumor-infiltrating cells, not tumor cells. The IDO mRNA correlated significantly with gene expression of interferon-gamma, tumor necrosis factor-alpha and interleukin-1beta. The recurrence-free survival rate of IDO-positive HCC patients was significantly higher than that of IDO negative HCC patients (P<0.05). CONCLUSIONS: These results suggest that IDO is a necessary enzyme for anticancer immune reactions of tumor-infiltrating cells. PMID- 14748881 TI - Plasma and urine levels of urinary trypsin inhibitor in patients with chronic liver diseases and hepatocellular carcinoma. AB - BACKGROUND AND AIM: Because urinary trypsin inhibitor (UTI) is synthesized by hepatocytes and excreted into the urine, plasma and urine levels of UTI may alter in liver diseases. However, there are few reports on the changes in these levels in chronic liver diseases and hepatocellular carcinoma (HCC). The aim of the present study was to evaluate the relationships between plasma and urine UTI levels and the severity of liver damage or progression of HCC in patients with chronic liver diseases and HCC. METHODS: Plasma and urine UTI levels were measured by a newly developed enzyme-linked immunosorbent assay in 16 patients with chronic hepatitis (CH), 19 patients with liver cirrhosis (LC) and 39 patients with HCC. RESULTS: Plasma UTI level exhibited a significant positive correlation with the urine UTI level. Plasma and urine UTI levels significantly decreased in LC patients compared with those of normal controls. In contrast, the plasma UTI level in HCC patients was higher than that in LC patients, but there was no difference between the groups in the urine UTI level. Plasma and urine UTI levels in LC and HCC patients were significantly correlated with the degree of liver damage according to the Child-Pugh classification. Although neither the plasma nor urine level of UTI in HCC patients were related to the clinical stage of HCC, both levels were closely associated with the level of protein induced by vitamin K absence or antagonist-II. CONCLUSIONS: The present findings indicate that the levels of plasma and urine UTI in patients with LC reflect the severity of liver damage. In HCC patients, these levels may also reflect progression of HCC, although further study is required. PMID- 14748882 TI - Role of L-carnitine in the prevention of acute liver damage induced by carbon tetrachloride in rats. AB - BACKGROUND AND AIM: Lipid peroxidation is the most important mechanism in the pathogenesis of acute liver damage with carbon tetrachloride (CCl4). L-carnitine may prevent lipid peroxidation and thus may protect against liver damage. In the present study we investigated the protective effect of L-carnitine in experimental acute liver damage induced by CCl4. METHODS: Fifty rats were allocated to five equal groups. The first group was the control (group 1), the second group received an intraperitoneal CCl4 injection for 3 days (group 2), and the third group received a 50 mg/kg subcutaneous L-carnitine injection for 4 days, beginning a day before CCl4 injection. The CCl4 injection was continued for 3 days in the concerned group (group 3). Group 4 was given a CCl4 injection for 7 days and group 5 received a 50 mg/kg subcutaneous L-carnitine injection for 8 days, beginning a day before CCl4 injection. This group continued to receive a CCl4 injection for 7 days. Rats in groups 2 and 3 were killed on the fifth day. Rats in groups 1, 4 and 5 were killed on the ninth day. Plasma and liver tissue malondialdehyde (MDA) levels, glutathione peroxidase (GSH-Px) activity and liver enzyme levels were studied. Histopathological investigations were conducted. RESULTS: Liver tissue MDA levels decreased significantly in group 3 compared to group 2 (P<0.001). Liver tissue MDA levels in group 5 decreased significantly in comparison to those of group 4 (P<0.001). Liver tissue GSH-Px activity in group 5 was significantly lower than that in group 4 (P<0.05). There were no significant differences between groups 3 and 4 regarding GSH-Px activity (P>0.05). Steatosis, inflammation and necrosis in group 3 were significantly reduced when compared to group 2 (P<0.01). Fibrosis development was not identified in groups 2 and 3. Steatosis in group 5 was significantly lower than that in group 4 (P<0.05) and there were no significant differences between groups 4 and 5 with regards to inflammation and necrosis (P>0.05). Mild fibrosis development was identified in groups 4 and 5 but the difference between the groups was not significant. CONCLUSION: It appears that L-carnitine has a protective effect in the early stage of experimental acute liver damage induced by CCl4. As the toxic effect or damage continues, its effect lessens. PMID- 14748884 TI - Gastrointestinal: late complication of percutaneous endoscopic gastrostomy. PMID- 14748883 TI - Hepatobiliary and pancreatic: fibropolycystic disease of the liver. PMID- 14748885 TI - Gastrointestinal: tapeworm infestation. PMID- 14748886 TI - ATP7b gene and Wilson's disease. PMID- 14748887 TI - Giant liver hemangioma in three sisters. AB - We describe a patient, in whom giant liver hemangioma (GLH) was found by ultrasound study in screening for hypertension. Two of her sisters also had GLH. One of them had become symptomatic and the hemangioma was successfully removed. The other sisters were carefully watched. Our patient didn't need any intervention in 4 years of follow-up. The pathogenesis of GLH is still unknown. Recent investigations show a role of the TIE receptor/angiopoietin system in vascular malformations. In literature we only found two other reports about a familial occurrence of liver hemangiomas. A genetic defect in familial GLH has not yet been identified. PMID- 14748888 TI - Inflammatory fibroid polyps of the stomach and Helicobacter pylori. PMID- 14748889 TI - Radiation-induced ulceration of the stomach secondary to hepatic embolization with radioactive yttrium microspheres in the treatment of metastatic colon cancer. PMID- 14748890 TI - Severe hepatotoxicity associated with a N-nitrosofenfluramine-containing weight loss supplement: report of three cases. PMID- 14748891 TI - Effects of levothyroxine suppressive therapy on bone mineral density in premenopausal women. AB - BACKGROUND: Levothyroxine (L-T4) is widely prescribed for treating thyroid disorders, but its effect on bone mineral density (BMD), is being debated. OBJECTIVES: We studied the effect of supraphysiologic doses of L-T4 on BMD in a group of premenopausal women. PATIENTS AND METHODS: We included 50 women (mean age=36.8 +/- 7.6 years) receiving L-T4 for at least 1 year for treating their benign cold thyroid nodules. Serum T3, T4, thyroid-stimulating hormone (TSH), parathyroid hormone (PTH), calcium, and phosphate and urine calcium and sodium levels of all patients were measured. Bone density at femoral neck and lumbar (L1 L4) regions was measured, using dual energy X-ray absorptiometry (DXA). RESULTS: No significant decrease was detected in the bone density of the subjects treated with L-T4 compared with the control group. CONCLUSION: L-T4 treatment for 1 year is not associated with increased risk of osteoporosis in premenopausal women but other potential adverse effects still need to be monitored in women receiving L T4 especially long-term. PMID- 14748892 TI - Permeability studies of alkylamides and caffeic acid conjugates from echinacea using a Caco-2 cell monolayer model. AB - BACKGROUND: Echinacea is composed of three major groups of compounds that are thought to be responsible for stimulation of the immune system--the caffeic acid conjugates, alkylamides and polysaccharides. This study has focussed on the former two classes, as these are the constituents found in ethanolic liquid extracts. OBJECTIVE: To investigate the absorption of these two groups of compounds using Caco-2 monolayers, which are a model of the intestinal epithelial barrier. RESULTS: The caffeic acid conjugates (caftaric acid, echinacoside and cichoric acid) permeated poorly through the Caco-2 monolayers although one potential metabolite, cinnamic acid, diffused readily with an apparent permeability (Papp) of 1 x 10(-4) cm/s. Alkylamides were found to diffuse through Caco-2 monolayers with Papp ranging from 3 x 10(-6) to 3 x 10(-4) cm/s. This diversity in Papp for the different alkylamides correlates to structural variations, with saturation and N-terminal methylation contributing to decreases in Papp. The transport of the alkylamides is not affected by the presence of other constituents and the results for synthetic alkylamides were in line with those for the alkylamides in the echinacea preparation. CONCLUSION: Alkylamides but not caffeic acid conjugates are likely to cross the intestinal barrier. PMID- 14748893 TI - Undertreatment of congestive heart failure in an Australian setting. AB - AIM: Guidelines for the management of patients with chronic heart failure have undergone change in recent years, with beta-blockers and spironolactone shown to reduce mortality when added to angiotensin converting enzyme (ACE) inhibitors, diuretics and digoxin. The aim of this study was to examine the therapeutic management of heart failure in patients admitted to Tasmania's three major public hospitals, with an assessment of the appropriateness of the therapy according to contemporary published guidelines. METHODS: An extensive range of clinical and demographic data was retrospectively extracted from the medical records of consecutive adult patients admitted to the medical wards of the hospitals with heart failure, either as a primary diagnosis or as a comorbidity, during a 6 month period in late 1999-early 2001. RESULTS: The 450 patients (57% females) had a mean age of 77.8 +/- 10.2 years, and were being treated with a median of seven drugs on hospital admission. The percentages of patients being treated with the major drugs of interest were: ACE inhibitors (50%), beta-blockers (22%), spironolactone (15%), digoxin (24%), loop diuretics (65%) and angiotensin-II receptor antagonists (8%). Almost 10% were taking a non-steroidal anti inflammatory agent. Less than one-half the patients who were receiving an ACE inhibitor were taking a target dose for heart failure. There were no significant differences in the pattern of drug use between the three hospitals. Underuse of heart failure medications was most pronounced in women and elderly patients. CONCLUSIONS: The data suggest that current guidelines for the treatment of heart failure are still not being reflected in clinical practice. The relatively low use of drugs shown to improve survival in heart failure is of concern and warrants educational intervention. PMID- 14748894 TI - Impact of pharmacist-conducted home visits on the outcomes of lipid-lowering drug therapy. AB - OBJECTIVE: To evaluate a pharmacist-conducted educational and monitoring programme, designed to promote dietary and lifestyle modification and compliance with lipid-lowering drug therapy, for patients with dyslipidaemia. METHODS: This was a prospective, randomized, controlled study. The participants were 94 adults, with 81 completing the study (intervention group: 39; control group: 42), with a cardiovascular-related diagnosis and discharged from hospital, between April and October 2001, on lipid-lowering drug therapy. Patients in the intervention group were visited at home monthly by a pharmacist, who educated the patients on the goals of lipid-lowering treatment and the importance of lifestyle issues in dyslipidaemia and compliance with therapy, assessed patients for drug-related problems, and measured total blood cholesterol levels using point-of-care testing. Patients in the control group received standard medical care. The main outcome measure was total blood cholesterol levels after 6 months, and an evaluation of patient and general practitioner satisfaction with the programme. RESULTS: There was no significant difference in baseline total blood cholesterol levels between the two groups. The reduction over the course of the study in cholesterol levels within the intervention group was statistically significant (4.9 +/- 0.7 to 4.4 +/- 0.6, P<0.005), whereas there was no change within the control group (P=0.26). At follow-up, 44% of the intervention group patients and 24% of the control group patients had cholesterol levels below 4.0 mmol/L (P=0.06). The reduction in total cholesterol in the intervention group should translate to an expected 21% reduction in cardiovascular mortality risk and a 16% reduction in total mortality risk--more than twice the risk reduction achieved in the control group. In addition, the programme was very well received by the patients and their general practitioners, by satisfaction questionnaire. CONCLUSION: A pharmacist-conducted educational and monitoring intervention improved the outcomes of lipid-lowering drug therapy. PMID- 14748895 TI - Adverse drug reaction teaching in UK undergraduate medical and pharmacy programmes. AB - OBJECTIVES: To assess the extent of teaching about the Committee on Safety of Medicine's Yellow Card scheme and adverse drug reactions within UK Schools of Medicine and Pharmacy. METHODS: A self-completed questionnaire sent to all heads of undergraduate schools of medicine and pharmacy within the UK. RESULTS: The majority of undergraduate syllabi feature the Yellow Card Scheme. Knowledge of the Yellow Card Scheme was assessed in 79% of pharmacy programmes and 57% of medical schools. Specialist speakers on the Yellow Card Scheme were infrequently used. Fewer than half of respondents provided students with a guide to reporting ADRs (43% pharmacy and 43% medical). There is some disagreement about the value of supplying students with printed material about the Yellow Card Scheme. Half of medical Schools did not think that supplying 'Current Problems In Pharmacovigilance' would be useful. CONCLUSIONS: It was found that in both medicine and pharmacy, courses differed substantially in teaching about the Yellow Card Scheme and adverse drug reactions (ADRs). There is scope for increased involvement of the Medicines and Healthcare products Regulatory Agency in undergraduate education, in line with recommendations from the National Audit Office. PMID- 14748896 TI - Andrographis paniculata in the symptomatic treatment of uncomplicated upper respiratory tract infection: systematic review of randomized controlled trials. AB - OBJECTIVE: To assess the efficacy of Andrographis paniculata in the symptomatic treatment of uncomplicated upper respiratory tract infection. METHODS: Systematic review of the literature and meta-analysis of randomized controlled trials. Mean difference in the reduction in symptom severity scores between treatment and control groups was calculated to obtain an overall estimate of effect. RESULTS: Four studies met our inclusion criteria and were reviewed. A total of 433 patients reported in three trials were included in the statistical analysis. Andrographis paniculata fixed combination with Acanthopanax senticosus was more effective than placebo. The mean difference was 2.13 points (95% CI 1.00-3.26 points, P=0.0002) on the symptom severity score. The difference in effects between A. paniculata and placebo was 10.85 points (95% CI 10.36-11.34 points, P<0.0001) in favour of A. paniculata. CONCLUSION: Current evidence suggests that A. paniculata extract alone or in combination with A. senticosus extract may be more effective than placebo and may be an appropriate alternative treatment of uncomplicated acute upper respiratory tract infection. PMID- 14748897 TI - A simple and rapid genotyping method for beta-2 receptor (beta2 AR) gene using allele specific multiplex PCR. AB - BACKGROUND: Polymorphism of the beta2-adrenergic receptor (beta2 AR) gene is an important determinant of the function of this receptor. It affects receptor down regulation and beta2-agonist responses. It has also been a focus of interest in attempts to elucidate the genetic basis of asthma, hypertension, obesity and cystic fibrosis. Several different techniques have been established to determine beta2 AR genotypes but none of these methods are simple enough to detect simultaneously all the five alleles of our research interest (Arg16/Gly16, 20T/C, Gln27/Glu27, -47T/C and Thr164/Ile164). OBJECTIVES: To develop a simple and rapid PCR based method for the simultaneous detection of five beta2 AR alleles. METHOD: DNA was extracted from whole blood using standard alkali lysis method. Primers specific at the 3' end for the polymorphic sites were designed. The nested allele specific PCR was optimized for reproducibility and specificity. Parameters investigated included concentrations of MgCl2, Taq polymerase, primers and annealing temperature, to produce specific bands of interest. DNA samples were selected at random and submitted for direct PCR sequencing. RESULT: The first PCR produced a fragment of size 710 bp, which was used as template for the subsequent duplex and triplex PCR to detect the mutation sites of the five alleles. The method was found reproducible and specific when used to genotype patients with bronchial asthma. The sequencing results confirmed the specificity of the PCR method. CONCLUSION: The simple and rapid method for the simultaneous detection of the five beta2 AR alleles is suitable for the study of beta2 polymorphism and its clinical consequences. PMID- 14748898 TI - Monitoring trough plasma concentrations of mycophenolate mofetil in patients with uveitis. AB - BACKGROUND: Mycophenolate mofetil (MMF) has been used successfully in patients with various forms of uveitis not responsive to other immunosuppressants. Nevertheless, for these patients neither recommendations for optimal dosage of MMF nor data concerning drug exposure of MMF are available. OBJECTIVE: To describe the results of the therapeutic drug monitoring (TDM) of MMF trough concentrations in a cohort of patients with uveitis, with the aim of optimizing the dosage of this drug, by maintaining a target concentration to achieve adequate immunosuppression with a minimal risk of therapeutic failure or toxicity. PATIENTS AND METHODS: This study describes the results of monitoring trough plasma concentrations of MMF in 12 patients with uveitis during a mean period of 21.4 months. Patients included one with Stevens-Johnson syndrome, one with Graves-Basedow's disease, one with Behcet's disease, one with idiopathic thrombocytopenic purpura and the rest with idiopathic uveitis. All patients were treated with steroids and additional therapy prior to treatment with MMF. RESULTS: Pharmacokinetic monitoring of mycophenolic acid (MPA) was performed with 108 trough plasma samples using an EMIT assay. Mean daily MMF dose was 24.5 +/- 6.3 mg/kg and mean trough MPA concentration was 2.9 +/- 1.9 microg/mL. Therapy was effective in 10 patients (83%). There were few side-effects: diarrhoea, excitement, agitation and cough that disappeared with daily dose reduction of MMF. CONCLUSIONS: MMF was effective in the majority of patients with uveitis with an acceptable profile of side-effects. TDM of MMF in patients with uveitis is clinically practicable and may help to optimize individual immunosuppressive therapy. We estimated that MMF dosages in the range of 0.5-1.5 g/day might be sufficient for treating uveitis and we recommend an initial target range of 2-4 microg/mL, which included 50% of our results. Randomized controlled trials are essential to confirm the efficacy of MMF in uveitis. PMID- 14748899 TI - Longer than recommended empiric antibiotic treatment of urinary tract infection in women: an avoidable waste of money. AB - CONTEXT: Current Israeli guidelines for the empiric treatment of uncomplicated urinary tract infection (UTI) in women recommend nitrofurantoin for 5 days. Some physicians nevertheless opt for ofloxacin, which should be prescribed for 3 days according to universally accepted guidelines. OBJECTIVE: To evaluate the economic consequences of longer than recommended durations of antibiotic therapy in the empiric treatment of uncomplicated UTI in women. DESIGN, SETTING AND PATIENTS: Data were derived from the electronic records of one of the four health maintenance organizations in Israel. The sample included all women aged 18-75 years who were diagnosed with acute cystitis or UTI from January 2001 to June 2002 and were empirically treated with antibiotics. Of the 7738 patients identified, 1138 received nitrofurantoin and 1054 ofloxacin. The excess expenditure accrued due to longer than recommended therapy with these drugs was evaluated. RESULTS: The rate of adherence was 22.23% for nitrofurantoin (95% CI=19.81%, 24.65%), and 4.08% for ofloxacin (95% CI=2.88%, 5.28%). The average excess expenditure per case was 5.78 USD (US Dollar) with ofloxacin and 3.43 USD with nitrofurantoin, resulting in an annual loss to the health maintenance organizations of approximately 19,000 USD. When extrapolated to the national population of 6.5 million, the loss due to inappropriate treatment of adult women is 190,000 USD. CONCLUSIONS: The lack of adherence to national and international guidelines with regard to the recommended duration of antibiotic treatment of UTI in women resulted in a significant and avoidable waste of health system resources. This study suggests that drug utilization analyses that concentrate solely on the choice of drug may be overlooking important information. PMID- 14748900 TI - Evaluation of once daily tobramycin dosing in critically ill patients through Bayesian simulation. AB - OBJECTIVE: To evaluate if once-daily dose (ODD) regimens of tobramycin attain pharmacodynamic goals using individualized pharmacokinetic monitoring of critically ill patients with creatinine clearance (Clcr) over 60 mL/min. METHODS: Fifty-one adult critically ill patients treated with intravenous tobramycin with ODD were included in the study. The effect of dosing using the proposed method was compared with a weight-based (7 mg/kg) dosing method. Pharmacokinetics parameters, peak concentration (Cpeak), minimum concentration (Cmin) and the time below the minimum inhibitory concentration (MIC) were estimated using Bayesian analysis. Pharmacodynamic parameters used to evaluate both dosing regimens were Cpeak/MIC ratio and, secondly, time below MIC (T< MIC). RESULTS: The median dose of tobramycin administrated in our hospital was too low for achieving pharmacodynamic goals. In contrast, the weight-based (7 mg/kg) method produced an adequate Cpeak/MIC ratio but an increase of the dose would not reduce the secondary pharmacodynamic index T60 mL/min achieved the Cpeak/MIC target values of 10. However in critically ill patients with Clcr>80 mL/min, T or =49 years (mean age 65 years). Serum creatinine and haemoglobin were measured using standard laboratory techniques. Creatinine clearance was estimated from serum creatinine, body weight, sex and age. RESULTS: Two hundred and seventy subjects (8.4%) had serum creatinine levels > or =125 micromol/L and estimated -creatinine clearances were <0.84 mL/s (50 mL/min) in 894 subjects (27.7%) and <0.50 mL/s (30 mL/min) in 120 subjects (3.7%). There was a strong association between reduced renal function and anaemia. Compared to those with serum creatinine <125 micromol/L, the age-adjusted relative risk (RR) of anaemia in women (haemoglobin <12.0 g/dL) with serum creatinine > or =125 micromol/L was 5.5 (95% confidence interval (CI) 2.9-10.7) and the RR of anaemia in men (haemoglobin < 13.0 g/dL) was 3.1 (95% CI 1.6-6.0). Estimated -creatinine clearance <50 mL/min was associated with a three-fold increased risk of anaemia in women and a five-fold increased risk in men. CONCLUSIONS: The results of the present study suggest that even modestly impaired renal function is associated with anaemia in older men and women. The possibility of renal impairment should be considered in the diagnosis and management of anaemia in people aged>50 years. PMID- 14748910 TI - Low prevalence of autoimmune diabetes markers in a mixed ethnic population of Singaporean diabetics. AB - BACKGROUND: Circulating antibodies to glutamic acid decarboxylase (GADab) and tyrosine phosphatase-like molecule IA-2 (IA-2ab) are major indicators for auto immune destruction of pancreatic islet cells. They identify a majority of Caucasians with type 1 diabetes and approximately 50% of Asians, providing evidence of an idiopathic aetiology in the latter. The present study investigated these autoantibodies in a mixed ethnic group. METHODS: Hospital clinic patients with clinically defined type 1 (n = 93) and type 2 (n = 300) diabetes and representing Singapore's major ethnic groups--Chinese, Indians and Malays--were studied. GADab and IA-2ab frequencies, and association of autoimmunity status with clinical and biochemical profiles were analysed. RESULTS: Radio immunoprecipitation assays detected either or both antibodies (seropositivity) in 41.9% of subjects with type 1 diabetes. GADab was detected in 36.6% and IA-2ab in 23.7% of type 1 diabetics. Prevalence of IA-2ab showed a reduction in frequency with disease duration (P = 0.026). In clinical type 2 diabetics, seropositivity was 10.0% with higher frequency in Malays (17.5%) than Chinese (9.7%) and Indians (4.5%). Multivariate analysis revealed that low fasting C-peptide was associated with seropositivity (odds ratio (OR) = 0.15; 95% confidence interval (CI) = 0.04 0.58). A significant relationship (OR = 13.5; 95% CI = 5.0-36.7) between insulin requirement and duration (>5 years) was also revealed. In patients with type 2 diabetes there was a trend of gradual progression to insulin dependency. However, there was considerable variation in body mass index between ethnic subgroups of type 2 diabetics, particularly for Chinese (mean (SD) = 26.0 (4.7)) and Malays (mean (SD) = 29.2 (5.9); P < 0.001). CONCLUSIONS: Presence of both antibodies in our mixed ethnic group of type 1 diabetes patients was much lower than in Caucasians. Significant numbers of patients were seronegative for antibodies. Influences due to ethnicity and adiposity would require further investigations. PMID- 14748911 TI - Audit of consultant physicians' reply letters for referrals to clinics in a tertiary teaching hospital. AB - BACKGROUND: Doctors referring patients to consultant physicians seek reply letters which both educate and assist in ongoing patient management. Highly desirable attributes in specialist letters include clearly stated and justified: (i) diagnostic formulations, (ii) management regimens, (iii) use of clinical investigations, (iv) prognostic statements, (v) contingency plans and (vi) follow up arrangements. AIM: To explicitly evaluate the quality of reply letters for new patients referred to clinics at a tertiary teaching hospital. METHODS: Letters were sampled from outpatient clinics of 10 different medical specialties at Princess Alexandra Hospital in Brisbane, Australia. Reply letters for new patient referrals between 1 August 2000 and 31 October 2000 were retrieved, from which data were abstracted to calculate the proportion of letters satisfying prespecified quality attributes. RESULTS: Of 297 new patient referrals, reply letters were retrieved for 204 (69%). Of these, 147 (72%) referrals were accompanied by a referral letter, mostly (113/147; 77%) from general practitioners. For 120 referrals involving diagnostic issues, 69 (56%) letters stated a diagnostic formulation. Of 114 letters recommending further clinical investigations, 61 (53%) described a rationale for such testing. In 125 cases where therapy was a key issue, 83 (66%) letters recommended changes to current treatment for which reasons were specified in 46 (55%) cases, and contingency plans provided in 13 (16%). Prognosis was mentioned in only 18 (9%) cases. Follow up arrangements were detailed in 123 (60%) letters. Assessments of patient understanding and likely adherence to therapy were stated in less than 15% of letters. CONCLUSIONS: Opportunities exist for improving quality of consultant physicians' reply letters in terms of greater use of problem lists, contingency plans, prognostic statements and patient-centred assessments, as well as more frequent enunciation of consultants' reasoning behind requests for further tests and changes to current management. Use of structured letter templates may facilitate more consistent inclusion of key information to referring doctors. PMID- 14748912 TI - Suspected white-tail spider bite and necrotic ulcers. AB - AIM: To describe the clinical features, investigation, diagnosis and treatment of ulcers attributed to white-tail (WT) spider bites or necrotic arachnidism. METHODS: The study was a prospective case series of patients referred to the Hunter Area Toxicology Service (a tertiary referral toxicology unit servicing a population of 500 000) with an ulcer or skin lesion that had been attributed to either a suspected WT spider bite or necrotic arachnidism. Eleven patients with skin lesions or necrotic ulcers were referred between January 2000 and June 2002. RESULTS: In two patients that were inpatients in other hospitals, investigation and follow up was not possible. In both cases there was no history of spider bite and Staphylococcus aureus was cultured. In nine patients, a diagnosis other than spider bite was made following appropriate investigation and follow up, including: (i) two cases of dermatophytoses, (ii) three staphylococcal infections, (iii) one case of pyoderma gangrenosum, (iv) one case of cutaneous polyarteritis nodosa, (v) one case of Nocardia braziliensis and (vi) one infected diabetic ulcer. There was only one case where the person recalled seeing a spider bite them, but the patient did not collect the spider for identification. The median time to diagnosis was 3 weeks (interquartile range: 3-9 weeks) and 3.5 years in one case. Appropriate treatment was initiated once the correct diagnosis was made and all cases resolved. CONCLUSIONS: In this series, all cases initially referred as WT spider bites or necrotic arachnidism were found to have alternative diagnoses with appropriate investigations. This demonstrates that spider bites are an unlikely cause of necrotic ulcers and that all ulcers should be properly investigated with bacterial, fungal and mycobacterial cultures and skin biopsy for histopathology. PMID- 14748914 TI - Evidence-based guide to slowing the progression of early renal insufficiency. AB - Early renal insufficiency (ERI), defined as a calculated or measured glomerular filtration rate (GFR) between 30 and 60 mL/min per 1.73 m2, is present in more than 10% of the adult Australian population. This pernicious condition is frequently unrecognised, progressive and accompanied by multiple associated comorbidities, including hypertension, renal osteodystrophy, anaemia, sleep apnoea, cardiovascular disease, hyperparathyroidism and malnutrition. Several treatments have been suggested to retard GFR decline in ERI, including blood pressure reduction, angiotensin-converting enzyme inhibition, angiotensin receptor antagonism, calcium channel blockade, cholesterol reduction, smoking cessation, erythropoietin therapy, dietary protein restriction, intensive glycaemic control and early intensive multidisciplinary patient education within a renal unit. In addition, specific interventions have been reported to be renoprotective in atherosclerotic renal artery stenosis, diabetic nephropathy, lupus nephritis and certain forms of primary glomerulonephritis. The present paper reviews the available published randomised controlled clinical trials and meta-analyses supporting (or refuting) a role for each of these therapeutic manoeuvres. PMID- 14748913 TI - Ongoing controversies in the pharmacological management of cancer pain. AB - Pain management remains a problem in advanced cancer. Despite the ready availability of effective analgesia and good evidence to support the prescription of medications, concerns continue over the safety of this practice. The aim of the present paper was to review often-raised questions when considering the use of opioids, especially in cancer pain, to ascertain the levels of evidence that already exist to support opioid-prescribing practice and to identify areas where further research is needed. PMID- 14748915 TI - Ross River virus disease in Australia: epidemiology, socioecology and public health response. AB - Societal and ecosystem change have a potentially profound impact of on human health and well-being. Alteration in the pattern of infectious diseases could be one of the most significant results of this process. Arboviral infections are a global public health issue with significant morbidity and mortality burden in the human population. Ross River virus (RRV) infection is the most common arboviral disease in Australia and some Pacific island nations. The present paper aims to illustrate the epidemiological and socioecological implications of RRV infection in Australia and to make recommendations for public health response to this disease. PMID- 14748916 TI - Genetic testing: a round table conversation. Part I. PMID- 14748917 TI - Cladribine followed by autologous stem-cell transplantation in progressive multiple sclerosis. AB - We studied the safety of autologous peripheral blood stem-cell transplantation (PBSCT) in four patients with progressive multiple sclerosis. Clinical and magnetic resonance imaging outcomes were secondary end-points. Cladribine administration preceded filgrastim-primed PBSC collection, aiming for lymphocyte depletion. Conditioning was with antithymocyte globulin and cyclophosphamide. The procedure was well tolerated, but without obvious clinical benefit. The study was terminated when other therapeutic options with lower morbidity became available. PMID- 14748918 TI - Internal carotid artery dissection: never too old. PMID- 14748919 TI - Prolonged survival after the diagnosis of metastatic hepatic epitheliod haemangioendothelioma. PMID- 14748920 TI - Biliary hamartomas (von Mayenburg complex): magnetic resonance imaging in a case report. PMID- 14748921 TI - Therapeutic drug monitoring of anti-tuberculous drugs. PMID- 14748925 TI - Induced ovulation and egg deposition in the direct developing anuran Eleutherodactylus coqui. AB - This study investigates ovulation and egg deposition behaviors in the anuran Eleutherodactylus coqui from Puerto Rico in response to stimulation with gonadotropin and gonadotropin releasing hormones. Five hormones were tested by injection over a range of doses, including mammalian LHRH, avian LHRH, fish LHRH, D-Ala6, des-Gly10 ethylamide LHRH and hCG. We report a low level of ovulation and egg deposition in response to all hormones, with the most complete and consistent results from the non-natural D-Ala6, des-Gly10 ethylamide LHRH derivative. To confirm the viability of eggs produced in this manner we performed in vitro fertilization experiments that resulted in the development of normal frogs. Reproductive behaviors in E. coqui are apparently not controlled by a mammalian form of LHRH as reported in other common laboratory anuran species. D-Ala6, des Gly10 ethylamide LHRH induces ovulation and deposition of mature and fertilizable eggs in E. coqui. PMID- 14748926 TI - Descriptive review of geographic mapping of severe acute respiratory syndrome (SARS) on the Internet. AB - From geographic mapping at different scales to location-based alerting services, geoinformatics plays an important role in the study and control of global outbreaks like severe acute respiratory syndrome (SARS). This paper reviews several geographic mapping efforts of SARS on the Internet that employ a variety of techniques like choropleth rendering, graduated circles, graduated pie charts, buffering, overlay analysis and animation. The aim of these mapping services is to educate the public (especially travellers to potentially at-risk areas) and assist public health authorities in analysing the spatial and temporal trends and patterns of SARS and in assessing/revising current control measures. PMID- 14748927 TI - Towards evidence-based, GIS-driven national spatial health information infrastructure and surveillance services in the United Kingdom. AB - The term "Geographic Information Systems" (GIS) has been added to MeSH in 2003, a step reflecting the importance and growing use of GIS in health and healthcare research and practices. GIS have much more to offer than the obvious digital cartography (map) functions. From a community health perspective, GIS could potentially act as powerful evidence-based practice tools for early problem detection and solving. When properly used, GIS can: inform and educate (professionals and the public); empower decision-making at all levels; help in planning and tweaking clinically and cost-effective actions, in predicting outcomes before making any financial commitments and ascribing priorities in a climate of finite resources; change practices; and continually monitor and analyse changes, as well as sentinel events. Yet despite all these potentials for GIS, they remain under-utilised in the UK National Health Service (NHS). This paper has the following objectives: (1) to illustrate with practical, real-world scenarios and examples from the literature the different GIS methods and uses to improve community health and healthcare practices, e.g., for improving hospital bed availability, in community health and bioterrorism surveillance services, and in the latest SARS outbreak; (2) to discuss challenges and problems currently hindering the wide-scale adoption of GIS across the NHS; and (3) to identify the most important requirements and ingredients for addressing these challenges, and realising GIS potential within the NHS, guided by related initiatives worldwide. The ultimate goal is to illuminate the road towards implementing a comprehensive national, multi-agency spatio-temporal health information infrastructure functioning proactively in real time. The concepts and principles presented in this paper can be also applied in other countries, and on regional (e.g., European Union) and global levels. PMID- 14748924 TI - Association studies for asthma and atopic diseases: a comprehensive review of the literature. AB - Hundreds of genetic association studies on asthma-related phenotypes have been conducted in different populations. To date, variants in 64 genes have been reported to be associated with asthma or related traits in at least one study. Of these, 33 associations were replicated in a second study, 9 associations were not replicated either in a second study or a second sample in the same study, and 22 associations were reported in just a single published study. These results suggest the potential for a great amount of heterogeneity underlying asthma. However, many of these studies are methodologically limited and their interpretation hampered by small sample sizes. PMID- 14748928 TI - Maternal concentration of polychlorinated biphenyls and dichlorodiphenyl dichlorethylene and birth weight in Michigan fish eaters: a cohort study. AB - BACKGROUND: Studies on maternal exposure to polychlorinated biphenyls (PCBs) reported inconsistent findings regarding birth weight: some studies showed no effect, some reported decreased birth weight, and one study found an increase in weights. These studies used different markers of exposure, such as measurement of PCBs in maternal serum or questionnaire data on fish consumption. Additionally maternal exposures, such as dichlorodiphenyl-dichloroethylene (DDE), which are related to PCB exposure and may interfere with the PCB effect, were rarely taken into account. METHODS: Between 1973 and 1991, the Michigan Department of Community Health conducted three surveys to assess PCB and DDE serum concentrations in Michigan anglers. Through telephone interviews with parents, we gathered information on the birth characteristics of their offspring, focusing on deliveries that occurred after 1968. We used the maternal organochlorine (OC) measurement closest to the date of delivery as the exposure. Although one mother may have contributed more than one child, serum concentrations derived from measurements in different surveys could vary for different children from the same mother. The maternal DDE and PCB serum concentrations were categorized as follows: 0 -< 5 microg / L, 5 -< 15 microg / L, 15 -< 25 microg / L, >or=25 microg / L. Using repeated measurement models (Generalized Estimation Equation), we estimated the adjusted mean birth weight controlling for gender, birth order, gestational age, date of delivery as well as maternal age, height, education, and smoking status. RESULTS: We identified 168 offspring who were born after 1968 and had maternal exposure information. We found a reduced birth weight for the offspring of mothers who had a PCB concentration >or=25 microg / L (adjusted birth weight = 2,958 g, p = 0.022). This group, however, was comprised of only seven observations. The association was not reduced when we excluded preterm deliveries. The birth weight of offspring was increased in women with higher DDE concentrations when controlling for PCBs; however, this association was not statistically significant. CONCLUSION: Our results contribute to the body of evidence that high maternal serum PCB concentration may reduce the birth weight in offspring. However, only a small proportion of mothers may actually be exposed to PCB concentrations >or=25 microg / L. PMID- 14748929 TI - Iris recognition as a biometric method after cataract surgery. AB - BACKGROUND: Biometric methods are security technologies, which use human characteristics for personal identification. Iris recognition systems use iris textures as unique identifiers. This paper presents an analysis of the verification of iris identities after intra-ocular procedures, when individuals were enrolled before the surgery. METHODS: Fifty-five eyes from fifty-five patients had their irises enrolled before a cataract surgery was performed. They had their irises verified three times before and three times after the procedure, and the Hamming (mathematical) distance of each identification trial was determined, in a controlled ideal biometric environment. The mathematical difference between the iris code before and after the surgery was also compared to a subjective evaluation of the iris anatomy alteration by an experienced surgeon. RESULTS: A correlation between visible subjective iris texture alteration and mathematical difference was verified. We found only six cases in which the eye was no more recognizable, but these eyes were later reenrolled. The main anatomical changes that were found in the new impostor eyes are described. CONCLUSIONS: Cataract surgeries change iris textures in such a way that iris recognition systems, which perform mathematical comparisons of textural biometric features, are able to detect these changes and sometimes even discard a pre enrolled iris considering it an impostor. In our study, re-enrollment proved to be a feasible procedure. PMID- 14748930 TI - Allomonal effect of breath contributes to differential attractiveness of humans to the African malaria vector Anopheles gambiae. AB - BACKGROUND: Removal of exhaled air from total body emanations or artificially standardising carbon dioxide (CO2) outputs has previously been shown to eliminate differential attractiveness of humans to certain blackfly (Simuliidae) and mosquito (Culicidae) species. Whether or not breath contributes to between-person differences in relative attractiveness to the highly anthropophilic malaria vector Anopheles gambiae sensu stricto remains unknown and was the focus of the present study. METHODS: The contribution to and possible interaction of breath (BR) and body odours (BO) in the attraction of An. gambiae s.s. to humans was investigated by conducting dual choice tests using a recently developed olfactometer. Either one or two human subjects were used as bait. The single person experiments compared the attractiveness of a person's BR versus that person's BO or a control (empty tent with no odour). His BO and total emanations (TE = BR+BO) were also compared with a control. The two-person experiments compared the relative attractiveness of their TE, BO or BR, and the TE of each person against the BO of the other. RESULTS: Experiments with one human subject (P1) as bait found that his BO and TE collected more mosquitoes than the control (P = 0.005 and P < 0.001, respectively), as did his BO and the control versus his BR (P < 0.001 and P = 0.034, respectively). The TE of P1 attracted more mosquitoes than that of another person designated P8 (P < 0.021), whereas the BR of P8 attracted more mosquitoes than the BR of P1 (P = 0.001). The attractiveness of the BO of P1 versus the BO of P8 did not differ (P = 0.346). The BO from either individual was consistently more attractive than the TE from the other (P < 0.001). CONCLUSIONS: We demonstrated for the first time that human breath, although known to contain semiochemicals that elicit behavioural and/or electrophysiological responses (CO2, ammonia, fatty acids) in An. gambiae also contains one or more constituents with allomonal (~repellent) properties, which inhibit attraction and may serve as an important contributor to between-person differences in the relative attractiveness of humans to this important malaria vector. PMID- 14748932 TI - Of genes and genomes - and dark matter. PMID- 14748931 TI - Attenuated allergic airway hyperresponsiveness in C57BL/6 mice is associated with enhanced surfactant protein (SP)-D production following allergic sensitization. AB - BACKGROUND: C57BL/6 mice have attenuated allergic airway hyperresponsiveness (AHR) when compared with Balb/c mice but the underlying mechanisms remain unclear. SP-D, an innate immune molecule with potent immunosuppressive activities may have an important modulatory role in the allergic airway response and the consequent physiological changes. We hypothesized that an elevated SP-D production is associated with the impaired ability of C57BL/6 mice to develop allergic AHR. METHODS: SP-D mRNA and protein expression was investigated during development of allergic airway changes in a model of Aspergillus fumigatus (Af) induced allergic inflammation. To study whether strain dependency of allergic AHR is associated with different levels of SP-D in the lung, Balb/c and C57BL/6 mice were compared. RESULTS: Sensitization and exposure to Af induced significant airway inflammation in both mouse strains in comparison with naive controls. AHR to acetylcholine however was significantly attenuated in C57BL/6 mice in spite of increased eosinophilia and serum IgE when compared with Balb/c mice (p < 0.05). Af challenge of sensitized C57BL/6 mice induced a markedly increased SP-D protein expression in the SA surfactant fraction (1,894 +/- 170% of naive controls) that was 1.5 fold greater than the increase in Balb/c mice (1,234 +/- 121% p < 0.01). These changes were selective since levels of the hydrophobic SP-B and SP-C and the hydrophilic SP-A were significantly decreased following sensitization and challenge with Af in both strains. Further, sensitized and exposed C57BL/6 mice had significantly lower IL-4 and IL-5 in the BAL fluid than that of Balb/c mice (p < 0.05). CONCLUSIONS: These results suggest that enhanced SP-D production in the lung of C57BL/6 mice may contribute to an attenuated AHR in response to allergic airway sensitization. SP-D may act by inhibiting synthesis of Th2 cytokines. PMID- 14748933 TI - Sterol autoxidation: from phytosterols to oxyphytosterols. PMID- 14748934 TI - Lack of dose-responsive effect of dietary phyto-oestrogens on transepithelial calcium transport in human intestinal-like Caco-2 cells. AB - Ca absorption has been shown to be unaffected by high luminal concentrations of two commonly consumed soyabean phyto-oestrogens (PO) (genistein and daidzein) in Caco-2 cells grown under oestrogen-depleted conditions. However, these compounds exhibit dose-dependent biphasic effects in some tissues, such as reproductive tissue and bone. Thus, in light of this biphasic activity, the effect of lower concentrations of genistein and daidzein on Ca absorption requires further investigation. Therefore, the aim of the present study was to investigate the effect of a range of concentrations of genistein and daidzein on Ca absorption in the human Caco-2 intestinal-like cell model. Caco-2 cells were seeded onto permeable filter supports and allowed to differentiate into monolayers. On day 21, the Caco-2 monolayers (n 12 per treatment), grown in oestrogen-deplete media, were then exposed to 10 nm-1,25-dihydroxycholecalciferol (1,25 (OH)2D3), or 1, 10 and 50 microm-genistein or -daidzein for 24 h. After exposure, transepithelial and transcellular transport of (45)Ca and fluorescein transport were measured. As expected, 1,25 (OH)2D3 stimulated Ca absorption in Caco-2 cells, by up regulating transcellular transport. Ca absorption was unaffected by either PO at luminal concentrations of 1, 10 or 50 microm, typical of intakes by Western and Asian populations as well as supplemental levels, respectively. The results of this model suggest that the proposed beneficial effects of supplemental levels of these PO compounds on bone mass in postmenopausal women more probably arise from direct effects on bone cells, and not by an indirect effect of these compounds on Ca absorption. PMID- 14748935 TI - Selenium and its relationship to cancer: an update. AB - Selenomethionine (Semet) is the major seleno-compound in cereal grains and enriched yeast whereas Se-methylselenocysteine (SeMCYS) is the major seleno compound in Se-accumulator plants and some plants of economic importance such as garlic and broccoli exposed to excess Se. Animals can metabolize both Semet and SeMCYS. Epidemiological studies have indicated an inverse relationship between Se intake and the incidence of certain cancers. Blood or plasma levels of Se are usually lower in patients with cancer than those without this disorder, but inconsistent results have been found with toenail-Se values and the incidence of cancer. There have been eight trials with human subjects conducted on the influence of Se on cancer incidence or biomarkers, and except for one, all have shown a positive benefit of Se on cancer reduction or biomarkers of this disorder. This is consistent with about 100 small-animal studies where Se has been shown to reduce the incidence of tumours in most of these trials. Se enriched yeast is the major form of Se used in trials with human subjects. In the mammary-tumour model, SeMCYS has been shown to be the most effective seleno compound identified so far in reduction of tumours. Several mechanisms have been proposed on the mechanism whereby Se reduces tumours. Even though SeMCYS was shown to be the most effective seleno-compound in the reduction of mammary tumours, it may not be the most effective seleno-compound for reduction of colon tumours. PMID- 14748936 TI - Responses of domestic fowl to excess iodine: a review. AB - Typically, poultry diets contain 1-2 mg I/kg, but higher concentrations are sometimes used to enhance the I content of eggs. In addition to an increased deposition of I in the yolk, other often adverse responses occur, especially at exceptionally high concentrations. Excess I in grower diets can prevent sexual maturation in male and female fowl, and in layer diets will progressively reduce egg production until, by about 2500 mg I/kg diet, ovulation is inhibited and egg production ceases. Most I accumulates in the thyroid gland, and it is likely that the mechanism responsible for these reproductive disorders involves a modification of thyroid hormone activity. Simultaneous with the declining rate of lay, feed intake declines, egg weight and yolk-cholesterol contents decrease and body weight increases. Whereas fertility is unaffected in female breeders, hatch of fertile eggs is reduced, hatch time extended and embryonic mortality and dead in-shell proportions increased. In contrast, male fertility is decreased because of an increased incidence of dead spermatozoa, although hatchability of eggs from normally fed hens is unaffected. All reproductive variables, together with feed intake and body weight, are normalised within about 7 d of returning to a diet with normal I levels. Excess I suppresses growth in meat-type chickens, but does not affect feed conversion efficiency. There are transient increases in plasma I and cholesterol concentration during excess I intake in all types of bird. The evidence for varying responses to different I sources is equivocal, but the consensus is that source is probably not important. PMID- 14748938 TI - DNA damage and susceptibility to oxidative damage in lymphocytes: effects of carotenoids in vitro and in vivo. AB - Reports on the effects of carotenoids are conflicting. The present paper examines similarities and differences from contiguous studies in vitro and in vivo. Single cell gel electrophoresis was used to measure the frequency of single-strand breaks (SSB) in the cell line MOLT-17 (as a model system) and human peripheral blood lymphocytes (PBL). MOLT-17 cells were supplemented with beta-carotene, lutein or lycopene at a range of concentrations (0.00-8.00 micromol/l) using a liposome delivery method. Uptake was dose-dependent. beta-Carotene concentration in the media had no effect on SSB in control cells, but incubation with lycopene or lutein (>2.00 micromol/l) increased the numbers of SSB in control cells. MOLT 17 DNA was less susceptible to oxidative damage (100 micromol H2O2/l, 5 min, 4 degrees C) following incubation with carotenoids between 0.50 and 1.00 micromol/l; at >1.00 micromol/l the effects were ambiguous. Apparently healthy male volunteers supplemented their habitual diets with lutein, beta-carotene or lycopene (natural isolate capsules, 15 mg/d, 4 weeks) in three independent studies, raising plasma concentrations to different extents. Lycopene and lutein had no effect on SSB in control PBL or following oxidative challenge. However, increased plasma beta-carotene was associated with more SSB in control cells whilst PBL DNA resistance to oxidative damage ex vivo was unaffected. These results suggest that the carotenoids are capable of exerting two overlapping but distinct effects: antioxidant protection by scavenging DNA-damaging free radicals and modulation of DNA repair mechanisms. PMID- 14748937 TI - The effect of a high-protein, high-sodium diet on calcium and bone metabolism in postmenopausal women and its interaction with vitamin D receptor genotype. AB - The influence of a high-Na, high-protein (calciuric) diet on Ca and bone metabolism was investigated in postmenopausal women (aged 50-67 years) who were stratified by vitamin D receptor (VDR) genotype. In a crossover trial, twenty four women were randomly assigned to a diet high in protein (90 g/d) and Na (180 mmol/d) or a diet adequate in protein (70 g/d) and low in Na (65 mmol/d) for 4 weeks, followed by crossover to the alternative dietary regimen for a further 4 weeks. Dietary Ca intake was maintained at usual intakes (about 20 mmol (800 mg)/d). Urinary Na, K, Ca, N and type I collagen cross-linked N-telopeptide (NTx; a marker of bone resorption), plasma parathyroid hormone (PTH), serum 25 hydroxycholecalciferol (25(OH)D3), 1,25-dihydroxycholecalciferol (1,25(OH)2D3), osteocalcin and bone-specific alkaline phosphatase (B-Alkphase) were measured in 24 h urine samples and fasting blood samples collected at the end of each dietary period. The calciuric diet significantly (P<0.05) increased mean urinary Na, N, K, Ca and NTx (by 19 %) compared with the basal diet, but had no effect on circulating 25(OH)D3, 1,25(OH)2D3, PTH, osteocalcin or B-Alkphase in the total group (n 24). There were no differences in serum markers or urinary minerals between the basal and calciuric diet in either VDR genotype groups. While the calciuric diet significantly increased urinary NTx (by 25.6 %, P<0.01) in the f+ VDR group (n 10; carrying one or more (f) Fok I alleles), it had no effect in the f- VDR group (n 14; not carrying any Fok I alleles). It is concluded that the Na- and protein-induced urinary Ca loss is compensated for by increased bone resorption and that this response may be influenced by VDR genotype. PMID- 14748939 TI - Evidence that dietary supplementation with carotenoids and carotenoid-rich foods modulates the DNA damage: repair balance in human lymphocytes. AB - Epidemiological evidence has shown that the habitual consumption of diets high in fruits and vegetables is associated with reduced risk of cancers. The challenge is to identify causal mechanisms of effect. The aim of the current study was to determine whether an increase in rate of removal of DNA single-strand breaks (SSB) following oxidative challenge could be provoked ex vivo in peripheral blood lymphocytes (PBL). The PBL were isolated from apparently healthy volunteers following dietary intervention with: (1) a mixed carotene capsule; (2) a daily portion of cooked minced carrots; (3) a matched placebo; (4) a portion of mandarin oranges; (5) vitamin C tablets. Single-cell gel electrophoresis was employed to measure baseline levels of SSB and DNA susceptibility to oxidative damage, and to monitor the number of SSB over 4 h, in both unchallenged and H2O2 treated PBL. The enzymatic capacity for repair of different types of DNA oxidative lesions was also measured using two related cell-free assays. There was no evidence that any of the dietary supplementation regimens altered baseline levels of SSB, provided any direct antioxidant protection or altered DNA repair capacity, with two exceptions: the number of SSB following exposure to H2O2 decreased more rapidly in PBL from volunteers given the mixed carotene capsules and repair patch synthesis activity in PBL increased from volunteers given the cooked carrots. These results suggest that carotenoids and carotenoid-rich foods can influence DNA damage:repair by modulation of discrete stages in the DNA repair mechanisms. PMID- 14748940 TI - Long-term vitamin C supplementation has no markedly favourable effect on serum lipids in middle-aged Japanese subjects. AB - Antioxidant vitamins have been reported to be associated with an improvement in blood lipid profiles, but results are not consistent. The present study was designed to determine whether long-term vitamin C supplementation could alter serum lipid concentrations in subjects who completed a 5-year population-based double-blind intervention trial. A total of 439 Japanese subjects with atrophic gastritis initially participated in the trial using vitamin C and beta-carotene to prevent gastric cancer. Before and upon early termination of beta-carotene supplementation, 134 subjects dropped out of the trial; finally, 161 subjects assigned to the high-dose group (500 mg vitamin C/d) and 144 subjects assigned to the low-dose group (50 mg vitamin C/d) were studied. No favourable effect of vitamin C supplementation on serum concentrations of total cholesterol, HDL- and LDL-cholesterol, and triacylglycerol was observed, although high-dose vitamin C supplementation increased serum vitamin C concentrations substantially. Among women, the mean change in serum triacylglycerol decreased (-0.12 mmol/l, 95 % CI 0.32, 0.09) in the high-dose group, but increased (+0.12 mmol/l, 95 % CI 0.03, 0.22) in the low-dose group. In addition, the mean change in serum triacylglycerol among women with hypertriacylglycerolaemia was statistically significant (-1.21, 95 % CI -2.38, -0.05) after high-dose vitamin C supplementation. The 5-year vitamin C supplementation had no markedly favourable effects on the serum lipid and lipoprotein profile. However, our present results do not preclude the possibility that vitamin C supplementation may decrease triacylglycerol concentrations among women with hypertriacylglycerolaemia. PMID- 14748941 TI - Antioxidant supplementation preserves antioxidant response in physical training and low antioxidant intake. AB - The present controlled-training double-blind study (supplemented (S) group, n 7; placebo (P) group, n 10) was designed to investigate whether an antioxidant mixture (Se 150 microg, retinyl acetate mg, ascorbic acid 120 mg, alpha tocopheryl succinate 20 mg) would allow overloaded triathletes to avoid adaptation failure in the antioxidant system [corrected]. Dietary intakes were recorded. The supplement of Se, and vitamins A and E provided 100 % of the French RDA. Four weeks of overloaded training (OT) followed 4 weeks of normal training (NT). After NT and OT, biological studies were conducted at rest and after a duathlon test (run 5 km, cycle 20 km, run 5 km). During the 4-week period of NT, blood levels of GSH levels increased in response to supplementation (P<0.05) and remained elevated during OT. Plasma glutathione peroxidase activity was significantly higher in the S group in all situations after NT and OT (P<0.01). The S group had increased erythrocyte Cu,Zn-superoxide dismutase activity in response to OT (P<0.05). Supplementation significantly reduced (P<0.05) the magnitude in duathlon-induced creatine kinase isoenzyme MB mass increase, which tended to be higher with OT (P=0.09). We conclude that the antioxidant mixture helped to preserve the antioxidant system during an OT-induced stress in subjects with initially low antioxidant intakes. Effects of supplementation during NT and/or OT are shown mostly by the alleviated muscle damage. The effects of the antioxidant mixture were observed for doses that can be provided by a diversified and well-balanced diet. The maintenance of normal nutritional status with regard to the antioxidant intake (Se, vitamins C and E) plays a key role in antioxidant adaptive effects during NT and OT. PMID- 14748942 TI - Oxyphytosterols are present in plasma of healthy human subjects. AB - The oxidised derivatives of phytosterols (oxyphytosterols) were identified in plasma samples from thirteen healthy human volunteers, using MS. All the samples contained noticeable quantities of (24R)-5beta,6beta-epoxy-24-ethylcholestan 3beta-ol (beta-epoxysitostanol) and (24R)-ethylcholestan-3beta,5alpha,6beta-triol (sitostanetriol) and also trace levels of (24R)-5alpha,6alpha-epoxy-24 ethylcholestan-3beta-ol (alpha-epoxysitostanol), (24R)-methylcholestan 3beta,5alpha,6beta-triol (campestanetriol) and (24R)-ethylch olest-5-en-3beta-ol 7-one(7-ketositosterol). The amounts of these oxyphytosterols in plasma varied from 4.8 to 57.2 ng/ml. There are two possibilities concerning the origin of these compounds. First, they could come from the small amounts of oxyphytosterols in food. Second, they could originate from the in vivo oxidation of phytosterols in plasma. Very few data actually exist concerning these compounds. Their identification in human samples suggests that further research is necessary in this field. PMID- 14748943 TI - A micronised, dispersible ferric pyrophosphate with high relative bioavailability in man. AB - Ferric pyrophosphate is a water-insoluble Fe compound used to fortify infant cereals and chocolate-drink powders as it causes no organoleptic changes to the food vehicle. However, it is only of low absorption in man. Recently, an innovative ferric pyrophosphate has been developed (Sunactive Fe trade mark ) based on small-particle-size ferric pyrophosphate (average size 0.3 microm) mixed with emulsifiers, so that it remains in suspension in liquid products. The aim of the present studies was to compare Fe absorption of micronised, dispersible ferric pyrophosphate (Sunactive Fe trade mark ) with that of ferrous sulfate in an infant cereal and a yoghurt drink. Two separate Fe absorption studies were made in adult women (ten women/study). Fe absorption was based on the erythrocyte incorporation of stable isotopes ((57)Fe and (58)Fe) 14 d after the intake of labelled test meals of infant cereal (study 1) or yoghurt drink (study 2). Each test meal was fortified with 5 mg Fe as ferrous sulfate or micronised, dispersible ferric pyrophosphate. Results are presented as geometric means. There was no statistically significant difference between Fe absorption from micronised, dispersible ferric pyrophosphate- and ferrous sulfate-fortified infant cereal (3.4 and 4.1 % respectively; P=0.24) and yoghurt drink (3.9 and 4.2 % respectively; P=0.72). The results of the present studies show that micronised, dispersible ferric pyrophosphate is as well absorbed as ferrous sulfate in adults. The high relative Fe bioavailability of micronised, dispersible ferric pyrophosphate indicates the potential usefulness of this compound for food fortification. PMID- 14748944 TI - Comparison of urinary monitoring, faecal monitoring and erythrocyte analysis of stable isotope labels to determine magnesium absorption in human subjects. AB - We have evaluated urinary monitoring and erythrocyte analysis to determine Mg absorption in human subjects as alternatives to the conventional technique of faecal monitoring by stable-isotope techniques. Ten healthy adults received 2.2 mmol (25)Mg in water, together with wheat bread, followed 15 min later by intravenous injection of 0.6 mmol (26)Mg (day 1). Brilliant blue and Yb (given on day 0 and day 1 respectively) served as qualitative and quantitative faecal markers. Urine was collected for 6 d after test meal intake. Complete collections of faeces were made until excretion of the second brilliant blue marker (given on day 7). Mg isotope ratios were determined by thermal ionisation-MS in urine and faeces and by inductively coupled plasma-MS in erythrocytes. Absorption was determined based on: (1) 6 d urine pools; (2) 24 h urine pools (collected 22-46 h after test meal intake); (3) erythrocytes from a blood sample drawn on day 14; (4) complete 6 d faecal pools; (5) faecal pools based on the first three consecutive stools after excretion of the first brilliant blue marker. Differences in mean Mg absorption (42 44 %) were statistically insignificant between techniques, except when based on 6 d urine pools for which the value was significantly lower (33 (sd 7) %, P=0.0003, ANOVA). The results indicate that Mg absorption can be determined from 24 h urine pools or erythrocytes obtained 14 d after test meal intake, an alternative method to the more time-consuming and labour-intense faecal monitoring. The choice of technique depends on practical and financial considerations. PMID- 14748945 TI - beta-Carotene-vitamin A equivalence in Chinese adults assessed by an isotope dilution technique. AB - The present study was carried out to determine the conversion factor of synthetic (2)H-labelled beta-carotene to vitamin A in Chinese adults by using a stable isotope dilution technique. Fifteen healthy volunteers aged 50-60 years were recruited for a 55 d experiment. The volunteers (nine males and six females) were each given a physiological dose of [(2)H8]beta-carotene (6 mg) in oil on the first day of the experiment, and a reference dose of [(2)H8]retinyl acetate (3 mg) in oil was given on the fourth day. Serum samples were collected at 0, 3, 5, 7, 9, 11, and 13 h on the first and the fourth days of the study, daily for 10 d, and then weekly from days 14 to 56. beta-Carotene and retinol were extracted from serum and isolated by HPLC, and their enrichments were respectively determined by using GC-electron capture negative chemical ionisation-MS and LC-atmospheric pressure chemical ionisation interface-MS. Four of the subjects exhibited beta carotene to vitamin A conversion factors of >29.0:1 on a molar basis and were termed 'poor converters'. In the eleven normal converters (seven males and four females), the calculated conversion factors of beta-carotene to retinol ranged from 2.0:1 to 12.2:1 with an average of 4.8 (sd 2.8):1 on a molar basis, and from 3.8:1 to 22.8:1 with an average of 9.1 (sd 5.3):1 on a weight basis. The 52 d post-intestinal absorption conversion was estimated to be about 30 % of the total converted retinol. PMID- 14748946 TI - Prenatal programming of angiotensin II type 2 receptor expression in the rat. AB - Exposure to undernutrition during fetal life has been proposed as an underlying cause of adult hypertension. Epidemiological studies demonstrating relationships between low birth weight and later CVD are supported by animal experiments indicating that manipulations of the maternal diet in pregnancy exert programming effects upon blood pressure control. Pregnant female Wistar rats were fed a control diet (n 13) or a low-protein diet (n 12) throughout pregnancy. At delivery all animals were fed the same standard laboratory chow diet. Analysis of nephron number in kidneys obtained from 4-week-old offspring showed that this was significantly (P<0.05) reduced in animals exposed to maternal protein restriction. At this age rats exposed to low-protein diets in utero had systolic blood pressures that were significantly greater than those of control animals (+23 mmHg, P<0.05). Administration of ascending doses of angiotensin II (1-40 ng/kg body weight intravenously) to 10-week-old anaesthetised female rats showed that the pressor response to the peptide was greater and more prolonged in animals exposed to low-protein diets in utero. Renal expression of mRNA for the angiotensin II type 1A receptor was similar in the two groups of rats, but low protein-exposed animals had significantly lower renal expression of the type 2 receptor (P=0.023). These results suggest that maternal nutritional status programmes expression of the renal angiotensin II type 2 receptor. This may play a key role in the impairment of renal development and the elevation of blood pressure noted in rats exposed to intra-uterine protein restriction. PMID- 14748947 TI - Intake of dietary iron is low in patients with Crohn's disease: a case-control study. AB - Patients with Crohn's disease (CD) often experience Fe deficiency (ID) and frequently alter their diet to relieve abdominal symptoms. The present study set out to assess whether patients with CD have dietary habits that lead to low Fe intakes and/or reduced bioavailable Fe compared with control subjects. Patients with asymptomatic CD were matched to controls (n 91/group). Dietary intakes of Fe and contributions from different food groups were compared using a 7 d food diary. Promoters and inhibitors of non-haem Fe absorption were investigated and a recently published algorithm was applied to assess bioavailable Fe. Fewer patients than controls met the reference nutrient intake for Fe (32% CD patients v. 42% controls). Overall, patients had significantly lower mean Fe intakes (by 2.3 mg/d) and Fe density (by 0.26 mg/MJ (1.1 mg/1000 kcal)) compared with controls (both P<0.001). Differences were mainly due to a preference among CD patients for low-fibre non-Fe fortified cereals, particularly breakfast cereals. In particular, control subjects had higher Fe intakes than matched CD subjects for men (P<0.001) and women less than 50 years (P=0.03). Intakes of both ascorbic acid (P<0.001) and phytic acid (P<0.01), but not animal tissue (P=1.0), were lower in patients with CD, but these had no overall effect on the predicted percentage of bioavailable Fe. Thus total bioavailable Fe was reduced in patients with CD due to lower intakes (P<0.01). Dietary Fe intakes are low in CD patients, which may contribute to an increased risk of ID and anaemia. Changing dietary advice may compromise perceived symptoms of the disease so the need for Fe supplementation should be carefully considered. PMID- 14748948 TI - Alcohol energy intake and habitual physical activity in older adults. AB - Alcohol forms a significant component of many diets and it supplements rather than displaces daily energy intake. Surprisingly, alcohol intake does not systematically increase body weight. The present study assessed whether a higher level of habitual physical activity in the daily environment is associated with a higher alcohol intake. Alcohol intake as part of total food intake was measured with a 7 d dietary record while at the same time physical activity was monitored with a tri-axial accelerometer for movement registration. Subjects were twenty women and twenty-four men, aged 61+/-5 years, of BMI 27.1+/-4.6 kg/m(2). Between subjects, there was a positive association between the level of habitual physical activity and alcohol intake (r 0.41; P<0.01). The subjects with higher alcohol intake had a higher activity level. On days with and days without alcohol consumption there was no difference in physical activity within subjects. In conclusion, it was shown that subjects with higher alcohol consumption are habitually more active. This may explain the lack of increasing body weight through additional energy intake from alcohol. PMID- 14748950 TI - Generalized equations for predicting body density of men. 1978. AB - 1. Skinfold thickness, body circumferences and body density were measured in samples of 308 and ninety-five adult men ranging in age from 18 to 61 years. 2. Using the sample of 308 men, multiple regression equations were calculated to estimate body density using either the quadratic or log form of the sum of skinfolds, in combination with age, waist and forearm circumference. 3. The multiple correlations for the equations exceeded 0.90 with standard errors of approximately +/-0.0073 g/ml. 4. The regression equations were cross validated on the second sample of ninety-five men. The correlations between predicted and laboratory-determined body density exceeded 0.90 with standard errors of approximately 0.0077 g/ml. 5. The regression equations were shown to be valid for adult men varying in age and fatness. PMID- 14748949 TI - Effect of equilibrated hydration changes on total body water estimates by bioelectrical impedance analysis. AB - The present study was performed to determine how equilibrated fluctuations in hydration affected the validity of bioelectrical impedance analysis (BIA) for body composition assessment. Total body water (TBW) expansion was induced by a 4 d endurance trial and the subsequent water loss was obtained over the recovery period. Twelve healthy men exercised on a cycle and treadmill alternately for 5 h/d over 4 d at moderate intensity. TBW, fat mass (FM) and fat-free mass (FFM) were assessed 3 d before the trial (control), and on the first and eighth day of recovery (R1 and R8 respectively). TBW was evaluated by (2)H dilution (TBW2H) as a reference method and by BIA (TBWBIA) at 100 kHz at the same time. TBW2H increased significantly between the control day and R1 by 1.87 (sd 1.11) litres (P=0.005) and TBWBIA by 1.38 (sd 1.56) litres (P=0.009). Both values returned to the control level on R8. For each period, TBW2H and TBWBIA did not differ significantly and were correlated (r(2) 0.85, P=0.0004 for the control day; r(2) 0.63, P=0.03 for R1; r(2) 0.75, P=0.02 for R8). Plasma Na concentration and osmolality did not differ between the control day, R1 and R8. FFM gain (1208 (sd 1983) g) and FM loss (-1168 (sd 906) g) between the control day and R1 were followed by a FFM decrease (-624 (sd 1281) g) and a FM increase (860 (sd 1212) g) between R1 and R8. As expected, these FFM and FM changes were significantly correlated with TBW variations. The present results provide evidence that BIA may be a useful method for estimating TBW when fluid shifts are equilibrated and electrolyte concentrations are unchanged. However, it is not a valid technique for assessing FM and FFM under these conditions. PMID- 14748954 TI - A comment on the European Society of Human Reproduction and Embryology/American Society for Reproductive Medicine consensus of the polycystic ovarian syndrome. AB - This commentary on the European Society of Human Reproduction and Embryology/American Society for Reproductive Medicine (ESHRE/ASRM) consensus on diagnosis, nomenclature and long-term health risks of the polycystic ovarian syndrome (PCOS) (conference in Rotterdam, Netherlands, March 2003) questions whether the preservation of the term PCOS sufficiently considers the modern aspects of the aetiology and pathogenesis of this complex syndrome. The misleading and simplified term PCOS, which comprises a variety of different entities, carries with it the risk of misinterpretation and under- and overestimation of symptoms, as well as of overlooking contraindications. Additionally, bias in future studies is pre-programmed. In this commentary, it is proposed that the term polycystic should be substituted with polyfollicular, and the term PCOS with functional hyperandrogenism, which is further subdivided into five additional groups. By using this new classification, most of the patients' dysfunctions belonging to the unspecifically termed PCOS are clearly defined, which leads on to the assumption that the diagnostic and therapeutic approaches required will improve, and the amount of bias in both basic and clinical studies will be reduced, respectively. PMID- 14748956 TI - Public health policy and infertility. AB - In assisted reproductive technology, three main areas are relevant from a health policy perspective: first, the technical excellence of the services rendered; second, a licensing and monitoring system to control all practitioners and the premises in which they work; and third, legislation to avoid abuse, prevent possible damage to the mother and the child and outlaw techniques unacceptable in a given cultural setting, including mechanisms to ensure the adherence by all to ethical and deontological principles. No one refuses the need to ensure that services offering assisted reproductive technologies adhere to technical excellence and everyone agrees that technical guidelines should exist and should be thoroughly enforced. When it comes to legislating about assisted reproductive techniques and, more specifically, to ruling in favour of limiting the use of existing technology to certain groups and not to others, or to ban altogether individual methods, views are sometimes totally divergent. In countries where legislation is fairly liberal, usually both physicians and the public view legislation with favour; conversely, where a number of restrictions exist, many complain that legislation impairs their ability to offer proper services. The solution seems to be a simple, well-defined, legislative act setting clear principles and providing for the creation of an Authority capable of resolving the myriad of individual problems that invariably arise from applying assisted reproductive techniques. Homo sum et nihil humanum a me alienum puto I am a human being and therefore cannot consider alien to me anything that helps another human being. PMID- 14748957 TI - Single embryo transfer - state of the art. AB - Every practitioner active in the field of assisted reproduction treatment is aware of the risks and complications related to twin and higher-order multiple pregnancies. Introduction of single embryo transfer (SET) into IVF/intracytoplasmic sperm injection (ICSI) is one of the possible ways of reducing the rate of twin pregnancy. Careful selection of patients, in combination with elective SET, has been shown to decrease the twin pregnancy rate while maintaining a stable ongoing pregnancy rate. The combination of a woman younger than 38 years of age, in her first or second IVF/ICSI cycle and with an embryo with a high implantation potential is the key to successful SET. This article will discuss embryo selection and patient selection and review the data published on SET. In the Centre for Reproductive Medicine at Middelheim Hospital, 39% of all transfers in 2002 were SET; the ongoing pregnancy rate remained stable at 30.6%. The twin (multiple) pregnancy rate declined to 11.7%. Particular attention should be drawn to the augmenting effect of the pregnancy rate of frozen-thawed cycles. Health economic data available so far subscribe the plea for SET. PMID- 14748959 TI - Recent developments in human oocyte, embryo and blastocyst vitrification: where are we now? AB - The target of any cryopreservation procedure should be to ensure high survival rates of living cells after thawing. Two important parameters determine the success of any cryopreservation protocol: the manner in which cells regain equilibrium in response to cooling, and the speed of freezing (cooling rate). Slow-rate freezing protocols result in the formation of ice crystals during cooling and warming. Vitrification, in which high cooling rates in combination with a high concentration of cryoprotectant are used, does not produce any ice crystals during cooling and warming. However, there is a practical limit to the attainable cooling speed, and also a biological limit to the concentration of cryoprotectant tolerated by the cells during vitrification. Although post-warming survival depends on the species, the developmental stage and the quality of the embryos being vitrified, it seems clear that vitrification methods are increasingly successful and might be a better method than slow cooling procedures in the field of cryobiology. Many of the potential problems and benefits underlying vitrification as a method of choice for embryo cryopreservation in clinical embryology will be discussed in this review. PMID- 14748960 TI - Micromanipulation of the human oocyte. AB - Intracytoplasmic sperm injection (ICSI) provides an excellent outcome in a consistent manner, and is therefore used worldwide as a routine procedure. Since its introduction, few modifications have been made to its methodology. Recently, a combination of ICSI with micro-hole drilling by laser (LA-ICSI) of the zona pellucida appeared to decrease oocyte degeneration rates and to improve embryo quality and implantation. Cytoplasmic transfer is a more recently introduced procedure where the objective is to improve the quality of patients' oocytes by transferring cytoplasm from a good quality donor oocyte, in cases where it is assumed that cytoplasm is compromised. Nuclear transfer, involving exchange of nuclei between donor and receptor oocytes, is still an experimental procedure, the objective being similar to cytoplasmic transfer in improving oocyte/embryo quality. A nuclear transfer procedure involving somatic cells for reproductive purposes should not be used in humans, for ethical and technical considerations. On the other hand, nuclear transfer for therapeutic purposes to obtain stem cells may be considered in respect of its unique potential in medicine. Finally, the most recently emerged new concept under investigation is the haploidization of somatic cells for the purpose of creating artificial gametes. PMID- 14748962 TI - Biochemical and functional characterization of the human zona pellucida. AB - A successful interaction between spermatozoa and the zona pellucida is critical for fertilization. This biological step reflects multiple sperm functions, including the acquisition and completion of capacitation, recognition and binding to specific zona pellucida receptors, and induction of the physiological acrosome reaction. The recognition of carbohydrate sequences by complimentary receptors has been demonstrated in gamete interaction in different animal species. It has been proposed that, in the human, sperm binding to the zona pellucida requires a 'selectin-like' interaction. The hemizona assay (a unique internally controlled bioassay that evaluates tight binding of human spermatozoa to the homologous zona pellucida) and advanced methods of carbohydrate analysis have been used to test this hypothesis. Compelling evidence exists to demonstrate that oligosaccharide recognition is also required for specific, tight human gamete binding. The induction of the acrosome reaction using the physiological inducers, i.e. the zona pellucida and progesterone, was also examined. It has also been demonstrated that there is a priming effect of the steroid on the acrosome reaction inducing capacity of the zona pellucida. These studies may allow for a better understanding of human gamete interaction in physiological and pathological situations. PMID- 14748963 TI - Epigenetic reprogramming in early embryonic development: effects of in-vitro production and somatic nuclear transfer. AB - A considerable proportion of offspring, in particular in ruminants and mice, born from nuclear transfer (NT)-derived and in-vitro-produced (IVP) embryos is affected by multiple abnormalities of which a high birthweight and an extended gestation length are the predominant features; a phenomenon that has been called 'large offspring syndrome' (LOS). The underlying mechanisms are largely unknown at present, but alterations of epigenetic modifications of embryonic and fetal gene expression patterns, primarily caused by alterations in DNA methylation are thought to be involved in this syndrome. In mammals, DNA methylation is essential for the regulation of transcription during development and differentiation. This review summarizes results from studies in which mRNA expression patterns from IVP and NT-derived embryos were compared with those of their in-vivo counterparts. Numerous aberrations have been found ranging from suppression of expression to de novo overexpression or more frequently to a significant up- or down-regulation of a specific gene. These observations emphasize the need for further epigenetic studies during preimplantation embryo development to gain insight into the molecular regulation correlated with an undisturbed embryonic and fetal development. Understanding molecular mechanisms will aid improvements in biotechnologies applied to early embryos in all species, including humans. PMID- 14748964 TI - Effect of culture environment on embryo quality and gene expression - experience from animal studies. AB - Recent studies comparing bovine oocyte maturation, fertilization and embryo culture in vivo and in vitro have demonstrated that the origin of the oocyte is the main factor affecting blastocyst yield, while the post-fertilization culture environment is critical in determining blastocyst quality, measured in terms of cryotolerance and relative transcript abundance, irrespective of the origin of the oocyte. Production of embryos in vitro, particularly when using an extended period of in-vitro culture, may predispose the embryo to phenomena such as the large offspring syndrome, which is likely to alter gene expression, particularly of imprinted genes. It is clear now that the post-fertilization culture environment has a profound effect on the relative abundance of gene transcripts within the embryo, and culture under suboptimal conditions for as little as 1 day can lead to perturbations in the pattern of expression. PMID- 14748965 TI - Assessment of mammalian embryo quality: what can we learn from embryo morphology? AB - Embryo morphology assessment, however imperfect it may be, is at present the most popular method for embryo selection prior to transfer, both in human and bovine assisted reproduction. A major difference between human and bovine embryos is the fact that in the latter, assessment of morphology is jeopardized by the opacity of the blastomeres, which is caused by lipid droplet accumulation. This opacity makes it difficult to assess nuclear and nucleolar morphology, aspects which can easily be evaluated in human zygotes or early cleaving embryos. However, recent research which focused on correlation between bovine embryo morphology and embryonic ultrastructure, gene expression and cryoresistance, has provided evidence that much more can be deduced from mere embryo morphology than previously thought. Morphological features such as colour of the blastomeres, the extent of compaction, timing of blastocyst formation and expansion and diameter of the embryo at hatching can be linked with embryo quality. On the other hand, cattle embryos of deviant chromosomal constitution or with aberrant genetic make up cannot be selected against by means of the current morphological techniques. Possible solutions include the visualization of bovine pronuclei at the zygote stage by means of ultracentrifugation or multiphoton laser scanning microscopy, and adjustment of genetic analysis in order to reconstruct embryo genetic make-up starting from the biopsy material. PMID- 14748966 TI - Embryo culture as a diagnostic tool. AB - Embryo culture can serve as a strong diagnostic tool, yielding useful information regarding the implantation potential of the human embryo. The information thus gained is useful for quality control of the embryology laboratory, success rates of the IVF/intracytoplasmic sperm injection (ICSI) programme, and counselling of the couple following failed cycles. Zygotes can be scored and zygote quality has been associated with further embryonic development and cleavage stage embryo quality. Early cleavage, cleavage rate, cleavage stage embryo grade and subsequent progression of these embryos to the blastocyst stage have all been shown to be individual and collective markers for the implantation-competent human embryo. This manuscript discusses embryonic markers of normality/quality throughout in-vitro culture starting with the zygote and ending with the blastocyst. PMID- 14748968 TI - Life after liquorice: the link between cortisol and conception. AB - Previous studies have reported both direct and indirect evidence correlating the probability of conception by IVF-embryo transfer with follicular metabolism of glucocorticoids by the enzyme 11 beta-hydroxysteroid dehydrogenase (11 betaHSD). To resolve disputes regarding the predictive value of measures of cortisol cortisone interconversion, this study has focused on compounds present in follicular fluid that can regulate enzyme activities within the ovary. Follicular fluid contains both hydrophilic compounds that can stimulate and hydrophobic components that can inhibit the oxidation of cortisol to cortisone by 11 betaHSD. These latest data indicate that: (i) cortisol:cortisone ratios in follicular fluid increase in proportion to the follicular content of the hydrophobic inhibitors of 11 betaHSD (r2 = 0.076; P < 0.01); (ii) the developmental potential of the oocyte and embryo, in terms of the probability of conception subsequent to embryo transfer, is positively correlated with follicular cortisol:cortisone ratios (12.9 +/- 0.3 in conception cycles versus 8.5 +/- 0.2 in non-conception cycles, P < 0.0001; odds ratio = 3364.48, P < 0.001); (iii) conception by IVF embryo transfer is associated with increased concentrations of the ovarian inhibitors of 11 betaHSD (odds ratio = 4.54, P < 0.01) but with decreased concentrations of the ovarian stimuli of 11 betaHSD (odds ratio = 0.18, P < 0.001). PMID- 14748969 TI - Quality management in the ART laboratory. AB - Unlike most medical laboratories that play a diagnostic role, laboratories for assisted reproductive technologies, ART laboratories, are involved in the treatment of infertile couples. Handling human gametes and producing human embryos in order to achieve much-sought pregnancies form the key tasks of an ART laboratory. The impact of the activities and the possible risks makes it necessary to ensure the safety and reproducibility of all methods. To achieve and maintain the highest level of patient care and the highest success rates, a quality management system should be implemented. Several guidelines, compiled by professional associations of ART experts, official international standards and quality management models have been developed and issued and can be applied. Irrespective of the choice, establishing a quality management system in an ART laboratory leads to a huge amount of additional work and requires a lot of investment in all kind of areas. However, due to the increased standardization and efficiency of all procedures as well as the improved transparency and traceability of all actions performed, the quality of service provided by the laboratory will improve substantially and the effort will be worthwhile. PMID- 14748971 TI - Total quality improvement in the IVF laboratory: choosing indicators of quality. AB - The purpose of this paper is to describe a programme of total quality improvement (TQI) within the IVF laboratory and to provide specific examples of indicators that could be used in such a TQI programme. Although TQI is sometimes confused with quality control (QC) and quality assurance (QA), there are major differences between the three quality plans: (i) QC is an activity designed to ensure that a specific element within the laboratory is functioning correctly; (ii) QA is a comprehensive programme designed to looks at a laboratory as a whole and to identify problems or errors that exist in an attempt to improve the entire process; (iii) TQI is also a comprehensive monitoring process designed not only to detect and eliminate problems, but also to enhance a laboratory's performance by exploring innovation and developing flexibility and effectiveness in all processes. Indicators used in a TQI plan should be objective, relevant to the laboratory, and measure a broad range of specific events or aspects of treatment that reflect the quality of care. Threshold values for each of the indicators should be based on how the specific protocols used in the laboratory impact the outcomes and the nature of the indicators on quality of care. PMID- 14748973 TI - United Nations and human cloning: a slender and fortunate defence for biomedical research. AB - Numerous biomedical scientists have contributed to the wide knowledge on the growth of preimplantation human embryos in vitro, now improving every aspect of the form of clinical care. These data were gained ethically in many countries, to open new vistas including the alleviation of infertility, preimplantation genetic diagnosis and stem cells, combined with some recent reports on human reproductive cloning. After detailed consultations with scientists, clinicians, ethicists and lawyers, many governments passed legislation permitting research under their own particular socially-defined conditions. Virtually all of them rejected reproductive cloning; a few have accepted therapeutic cloning. These legislatures saluted the many biomedical scientists striving to improve IVF and its derivatives, recognizing their immense medical potential. A motion recently placed before the United Nations then recommended a worldwide ban on all forms of human cloning. Proponents included the Vatican and many Roman Catholic countries, the USA and others. Opponents included Belgium, China, Japan, Brazil, UK, Germany and France. Mediation was achieved by Iran and other Muslim nations, and led to a motion passed by single vote for a two-year delay. This may be the first-ever proposal to ban worldwide a particular form of research. It sounds the alarm bells for further research. It raises questions about the UN being an appropriate forum for ethical decisions affecting the entire world and its future medicine. Large blocs of nations committed to particular religions and outlooks confronted each other, a situation in total contrast to the detailed and widespread consultations made by individual governments when deciding their own individual ethics. This event was clearly a narrow escape for free research as defined by each country's own jurisprudence. It also places research on human embryology and reproductive biomedicine into a more critical situation than before. Current liberalism in studies on assisted conception, clearly threatened by powerful adversaries, will have to be fortified to maintain the current impetus into newer forms of clinical care. PMID- 14748976 TI - Disclosure: a threat to egg donation. PMID- 14748974 TI - Gamete donation - where is the evidence that there are benefits in removing the anonymity of donors? a rejoinder to a patient's viewpoint. PMID- 14748977 TI - Plasticity following injury to the adult central nervous system: is recapitulation of a developmental state worth promoting? AB - The adult central nervous system (CNS) appears to initiate a transient increase in plasticity following injury, including increases in growth-related proteins and generation of new cells. Recent evidence is reviewed that the injured adult CNS exhibits events and patterns of gene expression that are also observed during development and during regeneration following damage to the mature peripheral nervous system (PNS). The growth of neurons during development or regeneration is correlated, in part, with a coordinated expression of growth-related proteins, such as growth-associated-protein-43 (GAP-43), microtubule-associated-protein-1B (MAP1B), and polysialylated-neural-cell-adhesion-molecule (PSA-NCAM). For each of these proteins, evidence is discussed regarding its specific role in neuronal development, signals that modify its expression, and reappearance following injury. The rate of adult hippocampal neurogenesis is also affected by numerous endogenous and exogenous factors including injury. The continuing study of developmental neurobiology will likely provide further gene and protein targets for increasing plasticity and regeneration in the mature adult CNS. PMID- 14748978 TI - Moderate controlled cortical contusion in pigs: effects on multi-parametric neuromonitoring and clinical relevance. AB - Over the last decade, routine neuromonitoring of ICP and CPP has been extended with new on-line techniques such as microdialysis, tissue oxygen (ptiO(2)), acid base balance (ptiCO(2), pH) and CBF measurements, which so far have not lead to clear-cut therapy approaches in the neurointensive care unit. This is partially due to the complex pathophysiology following a wide-range of brain injuries, and the lack of suitable animal models allowing simultaneous, clinically relevant neuromonitoring under controlled conditions. Therefore, a controlled cortical impact (CCI) model in large animals (pig) has been developed. After placement of microdialysis, ptiO(2), temperature and ICP catheters, an unilateral CCI injury (2.6-2.8 m/sec velocity, 9 mm depth, 400 ms dwell time) was applied and neuromonitoring continued for 10 h. CCI caused a rapid drop in CPP, ptiO(2) and glucose, whereas ICP, glutamate and lactate increased significantly. Most parameters returned to baseline values within hours. Lactate stayed elevated significantly throughout the experiment, but the lactate-to-pyruvate ratio (LPR) changed only slightly, indicating no severely ischemic CBF. Contralateral parameters were not affected significantly. Evaluation of brain water content and histology (12 h post-CCI) showed ipsilateral brain swelling by 5% and massive cell damage underneath the injury site which correlated with changes of ICP, CPP, glutamate, lactate, and ptiO(2) within the first hours post-CCI. Moderate controlled cortical contusion in pigs induced a complex pattern of pathophysiological processes which led to 'early' histological damage. Thus, this new large animal model will enable us to investigate the effect of therapeutic interventions on multi-parametric neuromonitoring and histological outcome, and to translate the data into clinical practice. PMID- 14748979 TI - Effect of decompression craniotomy on increase of contusion volume and functional outcome after controlled cortical impact in mice. AB - If, how, and when decompressive craniotomy should be used for the treatment of increased intracranial pressure after traumatic brain injury are widely discussed clinical subjects. Despite the large number of clinical studies addressing this issue, experimental evidence of a beneficial or detrimental role of decompressive craniotomy after brain trauma is sparse. Therefore, we investigated the influence of craniotomy on intracranial pressure, contusion volume, and functional outcome in a model of traumatic brain injury in mice. Male C57/Bl6 mice were craniotomized above the right parietal cortex and were subjected to controlled cortical impact injury. In control mice, the craniotomy was closed immediately after trauma, whereas in treated animals the craniotomy was left open. In control mice intracranial pressure (ICP) increased to a maximum of 23.7 +/- 3.1 mm Hg 6 h after trauma (p < 0.001), while in craniotomized animals, no ICP increase was observed. Twenty-four hours after trauma, the point in time of maximal lesion expansion, contusion volume in craniotomized mice was 40% smaller as compared to controls (18.3 +/- 2.0 vs. 30.2 +/- 3.5 mm(3), p < 0.04). Furthermore, craniotomized mice showed significantly improved motor function in a beam walking task (p < 0.04) and faster recovery of body weight after trauma (p < 0.02). Our results demonstrate that craniotomy blunts post-traumatic ICP increase, significantly reduces secondary brain damage and improves functional outcome after experimental TBI. Careful clinical evaluation of craniotomy as a therapeutic option after TBI in man may therefore be indicated. PMID- 14748980 TI - Relationship between intracranial pressure and cortical spreading depression following fluid percussion brain injury in rats. AB - Traumatic brain injury (TBI) is known to be accompanied by an increase in intracranial pressure (ICP) and in some cases, by spontaneous generation of cortical spreading depression (CSD) cycles. However, the role of CSD in the pathophysiology of cerebral contusion is still unknown. A multiparametric monitoring assembly was placed on the right hemisphere of the rat brain to evaluate ICP, DC potential, extracellular K(+), cerebral blood flow (CBF), and electrocorticogram in 27 rats during 5 h. Fluid percussion brain injury (FPBI) with the magnitude of the impact 2.9, 3.3, 4.1, and 5.0 atmospheres was induced to the left parietal cortex in animal groups A, B, C, and D, respectively. A slow increase in ICP was evident, and was pronounced in group C and especially in group D, where four of nine animals died during the monitoring. At the end of the 5 h experiment, the mean ICP levels were 6.75 +/- 2.87, 8.40 +/- 2.70, 12.75 +/- 4.03, 29.56 +/- 9.25, and the mean total number of CSD cycles was 2.00 +/- 1.41, 4.29 +/- 4.23, 11.71 +/- 13.29, and 20.11 +/- 19.26 in groups A, B, C, and D, respectively. The maximal level of intensity of CSD cycle generation after FPBI was obtained in group D, where almost constant activity was maintained until the end of the experiment. A significant coefficient of correlation between ICP level and total number of CSD cycles was found for all ICP measurements (r = 0.47-0.63, p < 0.05, n = 27), however more significant (p < 0.001) was the coefficient during the period of monitoring between 2 and 4 h after FPBI. Our results suggest that numerous repeating CSD cycles are typical phenomena in moderate and especially severe forms of FPBI. The rising number of CSD cycles under condition of an ICP level >/=20 mm Hg may demonstrate, with high probability, the unfavorable development of TBI, caused by growing secondary hypoxic insult. PMID- 14748981 TI - Differential concentration-dependent effects of prolonged norepinephrine infusion on intraparenchymal hemorrhage and cortical contusion in brain-injured rats. AB - Under clinical conditions catecholamines are infused to elevate cerebral perfusion pressure and improve impaired posttraumatic cerebral microcirculation. This, however, is associated with the risk of additional hemorrhage in the acute phase following traumatic brain injury. In the present study we investigated the dose-dependent effects of prolonged norepinephrine infusion on arterial blood pressure, blood glucose, and structural damage in brain-injured rats. At 4 h following induction of a focal cortical contusion (CCI), 40 rats were randomized to receive low (0.15), medium (0.3), or high dose (1 microg/kg/min) norepinephrine. Control rats were given equal volume of NaCl. Norepinephrine and NaCl were infused intravenously via Alzet osmotic pumps for 44 h. Mean arterial blood pressure (MABP), blood gases and blood glucose were determined before, at 4, 24, 48 h after CCI in repeatedly anesthetized rats (n = 28). Systolic arterial blood pressure (SABP) was measured using the tail cuff method in awake, restrained rats (n = 12). Cortical contusion and intraparenchymal hemorrhage volume were quantified at 48 h in all rats. MABP determined in anesthetized rats was only marginally increased. SABP was significantly elevated during infusion of medium and high dose norepinephrine in awake rats, exceeding 140 mm Hg. Medium and high dose norepinephrine significantly increased cortical hemorrhage by 157% and 142%, without increasing the cortical contusion volume. Low dose norepinephrine significantly reduced the cortical contusion by 44%. Norepinephrine aggravates the underlying brain damage during the acute posttraumatic phase. Future studies are needed to determine the least deleterious norepinephrine concentration. PMID- 14748982 TI - Differential effects of traumatic brain injury on the cytochrome p450 system: a perspective into hepatic and renal drug metabolism. AB - Traumatic brain injury is known to cause several secondary effects, one of which is altered drug clearance. Given the fact that patients who sustain TBI are subsequently treated with a variety of pharmacological agents for the purpose of either neuroprotection or physiological support, it is imperative to clarify changes in expression and/or activities of enzymes involved in clearing drugs. The mixed function oxidase system, which consists of cytochrome P450 and cytochrome P450 reductase, plays a vital role in phase I drug metabolism. This paper addresses the issue as to what extent TBI affects the levels and activity of various rat CYP450 subfamilies. Our results show that TBI induces tissue specific and time-dependent alterations. Total hepatic CYP450 content showed a biphasic response with a decrease seen at 24 h followed by an increase at 2 weeks. CYP450 reductase, in contrast, showed an opposite temporal profile. Immunoblot analyses and marker substrate metabolism demonstrated a clear decrease in hepatic CYP1A levels while a significant increase in kidney was seen at both 24 h and 2 weeks. A dramatic induction of CYP3A was evident at 2 weeks in liver, while no changes were noticed in CYP2B or CYP2D subfamilies. CYP4F subfamily showed induction in kidney only. Collectively, the data reveal the differential effects of TBI on hepatic and renal drug metabolism. PMID- 14748983 TI - Descending vasomotor pathways in humans: correlation between axonal preservation and cardiovascular dysfunction after spinal cord injury. AB - Cardiovascular dysfunction is common after cervical spinal cord injury (SCI) in humans. At least three spinal cord elements involved in cardiovascular control have been identified: descending vasomotor pathways (DVPs), sympathetic preganglionic neurons, and spinal afferents. However, little is known about the localization of the DVPs within the human spinal cord, which limits our understanding of the mechanisms of cardiovascular dysfunction after SCI. This study was undertaken to examine the association of cardiovascular abnormalities after SCI in humans with the severity of degeneration and axonal loss within the DVPs. A detailed chart review and histopathological examination of postmortem spinal cord tissue was conducted in individuals with cervical SCI (n = 7) and control individuals with an intact central nervous system (n = 5). Individuals with SCI were divided into group 1 (severe cardiovascular abnormalities) and group 2 (no/minor cardiovascular disturbances). The area of degeneration and the number of preserved axons within different areas of the spinal cord were quantitated using EMPIX imaging software. Two areas of possible localization of DVPs were investigated: area I, within the dorsal aspects of the lateral funiculus; and area II, within the white matter adjacent to the dorsolateral aspect of the lateral horn. Comparison of the extent of axonal degeneration in both SCI groups demonstrated that individuals in group 1 had more extensive axonal degeneration than those in group 2. The number of intact axons within areas I and II in individuals from group 1 was significantly lower than those from group 2 or control cases (p = 0.029; p = 0.028). The most dramatic axonal loss was observed within area I in individuals with cardiovascular dysfunction. We conclude that loss and degeneration of DVPs, which are localized within the dorsolateral aspects of the human spinal cord, contributes to abnormal cardiovascular control after SCI. This information adds to our knowledge of pathobiology of cardiovascular dysfunction after human SCI and may ultimately suggest novel therapeutic strategies as regenerative and reparative approaches become translated to the clinic. PMID- 14748984 TI - Synaptic blockade plays a major role in the neural disturbance of experimental spinal cord compression. AB - We analyzed dynamic processes of neural excitation propagation in the experimentally compressed spinal cord using a high-speed optical recording system. Transverse slices of the juvenile rat cervical spinal cord were stained with a voltage-sensitive dye (di-4-ANEPPS). Two components were identified in the depolarizing optical responses to dorsal root electrical stimulation: a fast component of short duration corresponding to pre-synaptic excitation and a slow component of long duration corresponding to post-synaptic excitation. In the directly compressed dorsal horn, the slow component was attenuated more (attenuated to 37.4 +/- 9.1% of the control) than the fast component (to 70.5 +/- 14.9%) (p < 0.01) at 400 msec after stimulation. Depolarizing optical responses to compression and to chemical synaptic blockade were similar. There was a regional difference between white matter (attenuated to 86.2 +/- 10.5%) and gray matter (to 72.6 +/- 10.4%) (p < 0.03) in compression-induced changes of the fast components; neural activity in the white matter was resistant to compression, especially in the dorsal root entry zone. Depolarizing optical signals in the region adjacent to the directly compressed site were also attenuated; the fast component was attenuated to 77.6 +/- 10.4% and the slow component to 31.8 +/- 11.3% of the control signals (p < 0.01). Spinal cord dysfunction induced by purely mechanical compression without tissue destruction was virtually restored with early decompression. We suggest that a disturbance of synaptic transmission plays an important role in the pathophysiological mechanisms of spinal cord compression, at least under in vitro experimental conditions of juvenile rats. PMID- 14748986 TI - [Pay attention to severe acute respiratory syndrome]. PMID- 14748985 TI - Enhanced vulnerability to NMDA toxicity in sublethal traumatic neuronal injury in vitro. AB - Traumatic brain injury causes neuronal disruption and triggers secondary events leading to additional neuronal death. To study injuries triggered by secondary events, we exposed cultured cortical neurons to sublethal mechanical stretch, thus eliminating confounding death from primary trauma. Sublethally stretched neurons maintained cell membrane integrity, viability, and electrophysiological function. However, stretching induced in the cells a heightened vulnerability to subsequent challenges with L-glutamate or NMDA. This heightened vulnerability was specifically mediated by NMDA receptors (NMDARs), as stretched neurons did not become more vulnerable to either kainate toxicity or to that induced by the Ca(2+) ionophore A23187. Stretch-enhanced vulnerability to NMDA occurred independently of endogenous glutamate release, but required Ca(2+) and Na(+) influx through NMDARs. Stretch did not affect the electrophysiological properties of NMDARs nor excitatory synaptic activity, indicating that specificity of enhanced vulnerability to NMDA involves postsynaptic mechanisms downstream from NMDARs. To test whether this specificity requires physical interactions between NMDARs and cytoskeletal elements, we perturbed actin filaments and microtubules, both of which are linked to NMDARs. This had no effect on the stretch-induced vulnerability to NMDA, suggesting that sublethal stretch does not affect cell survival through the cytoskeleton. Our data illustrate that sublethal in vitro stretch injury triggers distinct signaling pathways that lead to secondary injury, rather than causing a generalized increase in vulnerability to secondary insults. PMID- 14748987 TI - [An overview of treatment of SARS: urgency of prospective controlled clinical trial]. PMID- 14748988 TI - [Raise the realization of childhood rheumatic disease]. PMID- 14748989 TI - [Clinical characteristics and prognosis of 33 children with severe acute respiratory syndrome in Guangzhou area]. AB - OBJECTIVE: Since the outbreak of a highly contagious new pneumonia, atypical pneumonia or severe acute respiratory syndrome (SARS) occurred in Guangzhou area, 33 children with this syndrome were treated in the authors' hospital. The present study aimed to understand clinical characteristics and prognosis of pediatric SARS patients in Guangzhou area. METHODS: Clinical manifestations, laboratory and radiologic findings, therapeutic approaches and prognosis of the 33 children with SARS in Guangzhou area were analyzed. RESULTS: Of the 33 cases, 17 were males and 16 were females. The age was between 3 months to 13 years, and 3 - 12 years old patients accounted for 82%. Five (15%) cases had an evident history of contacting SARS patient before the symptoms occurred. Another 5 (15%) cases had a history that contacts of these patients (family members or friends) developed fever and/or cough later. The most common symptoms in this cohort were fever (100%) and cough (91%). Most of the cases had high fever, higher than 39 degrees C. Near half of the cases had nonproductive cough. The initial blood cells count showed that total white blood cell (WBC) count was (2.5 - 9.7) x 10(9)/L. In 22 (67%) cases the WBC count was < 5.0 x 10(9)/L, and in 10 (30%) WBC was (5.0 - 7.0) x 10(9)/L, in 18 cases most of the WBC were lymphocyte count. Chest radiograph showed patchy infiltrates, in 15 cases the changes were unilateral, and in 18 were bilateral. The radiologic changes developed fast, in some cases the changes progressed from one side to both sides. The opacity was absorbed slowly, significant absorption took in average two weeks. Elevated ALT was found in 3 cases and elevated CK-MB in 2 cases. Treatment included isolation, good ventilation of the ward, bed rest, supportive regimens, low volume oxygen inhalation, use of Chinese traditional medicine, antibiotics to prevent bacterial infection, and anti-inflammation therapy. All the patients recovered and discharged from hospital after a mean period of 10.0 +/- 3.8 days. CONCLUSION: SARS in children may have its own characteristics. The main clinical manifestations were high fever and cough while no severe toxic symptoms, nor respiratory failure was seen; few symptoms or signs suggesting involvement of systems other than respiratory system were seen. Chest radiograph showed uni- or bilateral asymmetric air-space infiltrates which could worsen quickly and were absorbed slowly. Though there were severe changes in the lung, the patients might not have corresponding symptoms or signs. The total white blood cell count in peripheral blood did not increase. All the patients studied had a favorable outcome after the combined treatment. PMID- 14748990 TI - [Diagnostic criteria for severe acute respiratory syndrome (SARS) in children (preliminary)]. PMID- 14748991 TI - [Protocols for diagnosis and treatment of SARS in children (preliminary)]. PMID- 14748992 TI - [National investigation program of systemic onset juvenile idiopathic arthritis in diagnosis, therapy and prognosis]. PMID- 14748993 TI - [Summary of the symposium on SARS in children]. PMID- 14748994 TI - [Summary of the 6th national symposium on pediatric immunology]. PMID- 14748995 TI - [Death of a child with AIDS]. PMID- 14748996 TI - [Association of HLA-A, B, and DR haplotypes with genotype in Chinese children with systemic lupus erythematosus]. AB - OBJECTIVE: Development of systemic lupus erythematosus (SLE) is not only associated with single loci of HLA gene, but also possibly related to certain haplotypes and genotypes of MHC. In the present study the authors explored association of HLA-A, B, DR haplotype and genotype with SLE in Chinese children, analyzed a large family with multiple SLE patients and genetic origin of SLE patients with HLA-DRB1 * 15, to discover the influence of linkage disequilibrium of HLA gene on SLE. METHODS: HLA-A, B, DR alleles were tested in 53 patients with SLE and 40 cases with their parents, 35 patients with SLE and HLA-DRB * 15 positive and 27 cases with their parents, a large family with SLE (18 members of three generations) and also 78 normal controls and 43 cases with their parents by microlymphocytotoxicity test and polymerase chain reaction - sequence specific primers (PCR-SSP). HLA-A, B, DR haplotype and genotype of SLE patients and controls were statistically calculated. The SLE patients with HLA-DRB1 * 15 and controls were analyzed for either the gene originated from the paternal or the maternal side. RESULTS: The variety of the haplotype in patient group (64/80) was less than that in control group (74/86). Only 9 haplotypes were found common between the patient group and control group. The frequency of the haplotype HLA A9B40DRB1 * 15 was significantly higher in patient group than that in control group (P < 0.05), RR was 10.726 0. Five members of the large family had haplotype A9B40DRB1 * 15, 2 of them were patients with SLE, 1 of them was positive for ANA and had Raynaud's phenomenon and 2 of them were normal. The rest of the family members were normal. The frequency of genotypes DRB1 * 09/DRB1 * 15 and DRB1 * 03/DRB1 * 15 in SLE group was significantly higher than that of control group (P < 0.05), RR was 7.772 7 and 14.272 7, respectively. The number of SLE children with gene HLA-DRB1 * 15 derived from their fathers was significantly higher than that of the children with the gene derived from the mothers. CONCLUSION: These findings suggested that haplotype HLA-A9B40DRB1 * 15 and genotypes HLA-DRB1 * 09/DRB1 * 15, HLA-DRB1 * 03/DRB1 * 15 were correlated with SLE. The predisposition of multiple loci seems to have an additive effect. The children with their gene HLA-DRB1 * 15 derived from their fathers might more easily suffer from SLE than those with the gene derived from their mothers, the underlying mechanism needs further studies. PMID- 14748997 TI - [Application of CD34+ autologous peripheral progenitor cell transplant in the treatment of children with refractory SLE]. AB - OBJECTIVE: Systematic lupus erythematosis (SLE) is a severe disease which affects the patient for many years and there is no radical cure for the disease. To explore a possible way to treat children with refractory SLE, the authors treated 2 children with grade III and IV lupus nephropathy for 5 years and 7 years respectively, mainly presented with persistent thrombocytopenia, proteinuria, pleural effusion with CD34(+) autologous peripheral progenitor cells transplantation. METHODS: Mobilized with G-CSF and collected with CS-3000 Cell Separator, passed through the CliniMacs CD34(+) cell selection device, the count of CD34(+) cells obtained reached 1.0 x 10(6)/kg and 1.7 x 10(6)/kg, respectively with the remaining of 2.0 x 10(5)/kg and 1.0 x 10(4)/kg of CD3(+) cells individually. The selected CD34(+) cells were frozen at -80 degrees C. The conditioning regimen consisted of cyclophosphamide [50 mg/(kg x day) for 4 days] plus ATG [Fresennius S 5 mg/(kg x day) for 3 days]. After 48 h treatment with cyclophosphamide, the frozen stem cells were infused back to the patients. RESULTS: Neutrophils recovered on 9 and 7 days after transplantation respectively in these 2 cases. Beginning from 15 days, the platelet count recovered and remained at over 100 x 10(9)/L. The sign of Cushing's syndrome disappeared completely 3 months after transplantation because discontinuing the steroid. One child's height had a 5 cm increase within 6 months after stopping steroid and this was the first height gain during the 7 years since she had had the disease. Till this paper was written, these 2 children were followed up for 13 months and 6 months, respectively, all the original symptoms and autoantibodies related to autoimmune disorders disappeared. But the cell-mediated immunity did not recover yet with the CD4(+) cell level still remained at a lower level. CONCLUSION: The effect of CD34(+) autologous peripheral progenitor cell transplantation on the children with refractory SLE was satisfactory so far, but the long-term effect remains to be confirmed by further studies on more cases. PMID- 14748998 TI - [Methylprednisolone and cyclophosphamide pulse therapy of severe systemic lupus erythematosus in children]. AB - OBJECTIVE: To study the effect of methylprednisolone (MP) and cyclophosphamide (CPA) intermittent intravenous pulse therapy and the clinical prognosis in children with severe juvenile onset systemic lupus erythematosus (JOSLE). METHODS: Thirty patients with JOSLE, diagnosed by clinical, laboratory or renal histological examinations, were enrolled in this study. Of the 30 patients, 27 were females and 3 were males, the mean age was (12 +/- 3) years, and 20 of the 22 patients who had undergone initial therapy had LN, and the clinical courses before being involved in the study were 3 to 12 months in nine patients. Twenty three of the 30 patients had clinical manifestations of renal damages, of whom 4 patients were proven by initial renal biopsy to have WHO type IV, 2 had type II,1 had type V and 1 had type III, and 7 patients had one or more manifestations of central nervous system, including chorea, seizures, cerebrovascular accident (CVA) and organic brain syndrome (OBS), simultaneously, 9 patients had nervous system symptoms without the clinical manifestations of renal damages, 3 patients had lupus crisis, 7 patients did not have any manifestations of renal or neurological damages. According to the protocol of the therapy, the patients were divided into 3 groups: group A (n = 18) patients were treated with MP plus CPA intermittent intravenous pulse for children with lupus nephritis, and with or without neuropsychiatric lupus erythematosus (NPLE), group B (n = 7) with pulsed doses of MP, followed by prednisone and tripterygium wilfordii hook f(T(whf)) for patients without renal or central nerves system damage, and group C (n = 5) with prednisone alone for patients with LN determined by clinical and laboratory features. The effects of those regimes and the clinical prognosis were observed. RESULTS: On short-term follow-up, the SLEDAI-2K (by weight of the renal damage) showed significant difference between group A and group B, but there was no significant difference at the 9th months of the therapy. The long-term follow-up lasted in average for (37.2 +/- 24.8) months. Nineteen patients were followed up for more than 18 months. At the end of follow-up, the mean age was 14 to 19 years. There was no difference on the effect of both group A and group B, and no frequent infections were seen, ANAs were negative and SLEDAI-2K = 0-point in two patients of each group 12 months after discontinuation of the therapy. Four patients in group C died within 18 months. CONCLUSION: The immunosuppressive regimen MP + CPA in patients with severe JOSLE and MP + prednisone + T(whf) in patients without major organs damage were superior to the regimen of prednisone alone. PMID- 14748999 TI - [Treatment of patients with juvenile rheumatoid arthritis with combination of leflunomide and methotrexate]. AB - OBJECTIVE: To evaluate the efficacy and safety of the combined therapy with leflunomide and methotrexate in the patients with juvenile rheumatoid arthritis (JRA). METHODS: Forty patients with active polyarthritis JRA were divided into 2 groups. Group 1 (n = 21) received leflunomide tablet (1 mg/(kg x day) on days 1 - 3; then [(0.2 - 0.4) mg/kg per day] plus methotrexate (0.3 mg/kg i.v. every two weeks till clinical remission, then oral tablet 0.2 mg/kg weekly). Group 2 received the same doses of methotrexate in the same way. Permitted concomitant drugs included stable doses of NSAIDs and a low dose of prednisone during the course of treatments. The clinical assessments included the number of tender and swollen joints, tender articular index, swollen articular index, general articular function score, parents and physician's evaluation score, erythrocyte sedimentation rate, serum C-reactive protein and rheumatoid factor. Drug safety was assessed by observing the reaction of mucous membrane, skin, gastrointestinal tract, nervous system, hematologic changes, liver and renal function. Statistical comparison between two groups was performed by using analysis of variance, t test and chi(2) test. RESULTS: Efficacy and safety was assessed at 12th and 26th week. Average improvement rate of leflunomide plus methotrexate group at 12th week and 26th week was respectively 39.6% and 71.9%; while that of control group was 27.5% and 49.5%, i.e., there was significant difference between the two groups (P < 0.01). Average remission rate of leflunomide plus methotrexate group at 12th week and 26th week was respectively 4.76% and 38.10%; while that of control group (methotrexate only) was respectively 0, 0. The clinical improvement in the group treated with leflunomide plus methotrexate was significantly greater than control group (P < 0.01). There was no significant difference (9.5% v 5.3%) in occurrence rate of side effects between the two groups. Side effects included leucocytopenia and raised aminotransferase. They were mostly mild and tolerable. CONCLUSION: The effect of the leflunomide and methotrexate therapy in patients with active JRA was better than methotrexate alone. The combination therapy with leflunomide and methotrexate was safe and well tolerated. PMID- 14749000 TI - [A case of eosinophilic meningitis caused by cysticercosis of brain]. PMID- 14749001 TI - [Etiology of severe acute respiratory syndrome]. PMID- 14749002 TI - [Human metapneumovirus may associate with acute respiratory infections in hospitalized pediatric patients in Beijing, China]. AB - OBJECTIVE: A new respiratory virus, human metapneumovirus (HMPV) was recently identified by scientists in the Netherlands first and then in a few other countries. To investigate if this newly discovered virus is associated with the acute respiratory infections in pediatric patients in Beijing, tests were developed to detect HPMV gene fragments from nasopharyngeal aspirates collected from infants and young children hospitalized for acute respiratory infections from November 2002 to March 2003. METHODS: The HMPV was screened by reverse transcription-polymerase chain reaction (RT-PCR). RNAs were extracted by Trizol from 247 specimens which had been determined as negative for conventional respiratory viruses including RSV, influenza A and B, parainfluenza I, II, III and adenovirus by indirect immunofluorescence test as well as virus isolation. The HMPV RNAs were detected by reverse transcription tests using random primer and M-MLV reverse transcriptase followed by PCR using the primers designed from the published sequence of the N protein-encoding gene from the first HMPV identified in the Netherlands. PCR products were visualized by 1.2% agarose gel electrophoresis. Selected positive PCR products were sequenced and the sequences of the nucleotides and deduced amino acids were compared with those in the GenBank. RESULTS: Among those 247 specimens negative for common respiratory viruses, 74 (30.0%) showed the predicted 213 bp PCR products in agarose gel. Most of clinical diagnoses for these 58 patients were pneumonia (36, 48.6%), bronchiolitis (21, 28.4%), and bronchitis and asthma in some patients. Nearly 90 percent of positive specimens were from patients under 2 years of age. Ten out of 74 amplicons were randomly selected for sequence analysis. When compared with the sequences in the GenBank, the nucleotide sequences of these 10 amplicons shared high homology only with those of HMPVs. The nucleotide sequence identities of these 10 samples with those from the Netherlands and Canada were 87% - 99%. When compared with the nucleotide sequence from the first reported strain by Van den Hoogen (strain HMPV 00-1), the sequence identities of these 10 fragments ranged from 88.7% to 99.1%. Among the 10 amplicons from the specimens, the nucleotide identities were 87.3% - 100%. One of the 10 amplicons (No. 1816) shared lower identity with others (87.3% - 89.7%), whereas the other 9 shared higher identities (95.8% - 100%) with each other. The comparison of amino acids showed that these 10 amplicons showed high homology (95.8% - 100%). Again, amplicon No.1816 shared lower homology (95.8% - 97.2%) with others, whereas the other 9 shared higher homology (98.6% - 100%). The amino acid homology between No.1816 and HMPV 00-1 was 95.8%, whereas that of the other 9 with HMPV 00-1 was 98.6% - 100%. CONCLUSION: These data suggested that some of acute respiratory infections in pediatric patients in Beijing area are related to the newly identified human metapneumovirus. The HMPV circulating in Beijing may have different genotypes. PMID- 14749003 TI - [Filariae in vertebral canal, report of a case]. AB - OBJECTIVE: To confirm diagnosis of a special case with chief complaints of abdominal pain and dyskinesia of lower extremities. METHODS: The clinical symptoms, signs, MRI, pathological findings and the results of blood test for microfilaria were analyzed. RESULTS: The patient was a 6-year old girl who had abdominal pain for 10 days dyskinesia of lower extremities for 6 days accompanied by difficulty in urination and defecation. There was tenderness on T7-9 spinous process, sensory dullness below the umbilicus. Babinski's and Oppenheim's sign were bilaterally positive, and ankle clonus was positive. MRI showed space occupying change in the vertebral canal at T7-9 level. The mass of 2 cm x 1 cm x 1 cm size was removed by surgical operation and histopathological study showed obvious fibrous tissue proliferation accompanied by eosinophil, lymphocyte and neutrophil infiltration around a worm-like structure. Night time blood test performed at 23:00 confirmed the presence of microfilaria. CONCLUSION: The diagnosis of filariae in vertebral canal could be confirmed. PMID- 14749004 TI - [Role of free radicals in brain edema induced by endotoxin in infant rats and the therapeutic effect of dexamethasone and IVIG]. AB - OBJECTIVE: Glucocorticoid is considered as an effective drug for prevention and treatment of brain edema and reducing the blood-brain barrier (BBB) permeability. Intravenous immunoglobulin (IVIG) is frequently used to treat neurological diseases with immune abnormality, its function and potential mechanism on brain edema have not been reported. In this study, the roles of the total hydrosulfide group (TSH), non-protein hydrosulfide group (NPSH) and malondialdehyde (MDA) in etiology of the endotoxin brain edema in infant rats and the interfering effects of dexamethasone (DEX) and IVIG were investigated. METHODS: In 35 infant rats, 10 mg/kg lipopolysaccharide (LPS) was intraperitoneally injected. The same volume of normal saline was injected to 24 control rats. Ten mg/kg DEX and 400 mg/kg IVIG were intravenously injected respectively to 36 and 24 infant rats instantly following LPS injection. The TSH, NPSH and MDA concentrations and the brain Evans blue contents were detected at different time in the brain tissue. The brain water content was measured by drying method. RESULTS: The brain water, EB and MDA contents after endotoxin injection were significantly higher than those of control group, while the brain TSH, NPSH content were significantly lower than those of control group (P < 0.05 or P < 0.01); After treatment with DEX or IVIG, the brain EB, MDA and water content significantly decreased with the peak at 6 h (P < 0.05 or P < 0.01), TSH and NPSH significantly increased compared with LPS group. However, the NPSH content in IVIG treatment group did not change significantly (P > 0.05). CONCLUSION: Free radicals play a role in the brain edema induced by LPS in infant rats. The primary results suggested that DEX and IVIG have therapeutic effect for the endotoxin-induced brain edema by affecting the free radicals. PMID- 14749006 TI - [Report of a case with Schwardz-Jampel syndrome]. PMID- 14749005 TI - [Prader-Willi syndrome and genomic imprinting]. AB - OBJECTIVE: Prader-Willi syndrome (PWS) is an example of a human genetic disorder that involves imprinting genes on the proximal long arm of chromosome 15 and SNRPN gene as a candidate gene for this syndrome. The purpose of this study was to show the molecular genetic defects and genomic imprinting basis in Chinese PWS patients and to evaluate the clinical applications of a differential diagnostic test for PWS. METHODS: Fluorescence in situ hybridization (FISH) and methylation specific PCR (MSPCR) techniques were applied for 4 clinically suspected PWS patients. Using three probes, including SNRPN probe for identification of the critical locus in PWS region, D15Z1 and PML control probes for identification of the 15p arm and 15q arm, the authors detected the deletions 15q in PWS. MSPCR was based on sodium bisulfite treatment of DNA and PCR primers specific for the maternal and paternal allele. RESULTS: When hybridized with mixed probes, it was found in 2 patients that the central specific signal was absent, but both the flanking control signals were retained, indicating SNRPN gene deletion of chromosome 15q11-13. Bisulfite-modified DNA from all PWS children amplified with methylated allele-specific primer pair showed only maternal 131bp PCR product, indicating the maternal uniparental disomy (UPD15). CONCLUSION: Genomic imprinting plays an important role in the molecular pathogenesis of PWS that caused by paternal microdeletions of 15q11-q13 or maternal UPD of chromosome 15. The basic defect seemed to be an absence of function of PWS genes that are normally expressed only from the paternal chromosome 15. MSPCR is a rapid and simple PCR-based assay compared with other cyto-molecular tests and its results were consistent with the clinical diagnosis of PWS, so it seems to be a reliable diagnostic method for PWS patients who show abnormal methylation at SNRPN. The genetic differential tests for PWS are important in determining familial recurrence risk. PMID- 14749007 TI - [Change of neurokinin A plasma level in asthmatic children]. AB - OBJECTIVE: Chronic inflammation of airway in bronchial asthma is caused by many complicated elements. Recently, a close attention has been paid to the neurogenic inflammation in airway which is mediated by sensory neuropeptides secreted by sensory nerve in the lung. Neurokinin A (NKA) is an important sensory neuropeptide leading to neurogenic inflammation in airway. Experimental studies showed that NKA has a close relation to asthma. The purpose of the present study was to investigate the changes of NKA in plasma of asthmatic children and possible relationship between sensory neuropeptides and asthma in children. METHODS: Thirty-five children with bronchial asthma were studied; 16 of the cases were < 3 yrs and 19 were >or= 3 yrs. Eighteen of the cases had severe asthma and 16 had mild asthma. None of the subjects was treated with glucocorticoid within 3 days before the study started; 15 healthy children without history of asthma or family history of asthma were enrolled as control subjects. Plasma was collected from each case during acute attack of asthma and their clinical remission of the asthmatic children. After purifying with SEP-pak C(18), NKA content was detect by enzyme-linked immunosorbent assay (ELISA) as instructed by the manufacturer of the NKA Kit (NKA unit: ng/L). RESULTS: (1) The content of plasma NKA of asthmatic children was significantly higher at the asthma attack (256 +/- 153) than that at their clinical remission stage (70 +/- 66; q = 9.497, P < 0.01) and than that of the normal control group (38 +/- 6; q = 8.599, P < 0.01); no significant difference in plasma NKA was found between the clinical remission stage and the normal control group (q = 1.245, P > 0.05). (2) There was a significant positive correlation between the asthmatic clinical state and the levels of plasma NKA; the contents of plasma NKA at the stage of acute attack in severe asthma (296 +/- 170) were significantly higher than those of the mild asthmatic children (190 +/- 99; q = 3.77, P < 0.05). CONCLUSIONS: The contents of plasma NKA were significantly higher during the asthma attack stage of children, and the higher was the level of NKA, the more severe the attack.; with asthma remission, the contents of plasma NKA decreased to normal; the contents of plasma NKA has a close relation to the asthmatic children. PMID- 14749008 TI - [Effects of hypothermia on cardiac function in neonates with asphyxia]. AB - OBJECTIVE: To study the effects of hypothermia on cardiac function in neonates after birth asphyxia. METHODS: Fifty term newborns with Apgar score < 5 at 5 minutes were randomly divided into no cooling group (normothermia group, NG; rectal temperature = 36.5 degrees C +/- 0.5 degrees C, n = 27) and cooling group (hypothermia group, HG; rectal temperature = 34.5 degrees C +/- 0.3 degrees C, nasopharyngeal temperature = 34.0 degrees C +/- 0.5 degrees C, n = 23). The selective head cooling was applied to maintain nasopharyngeal temperature at 34 degrees C for 72 h in hypothermia group. Systolic and diastolic function was detected at the end of treatment by echocardiogram. RESULTS: (1) The heart rate was obviously decreased during the hypothermia treatment, and there was a significant difference between HG and NG [(103 +/- 15) bpm vs. (126 +/- 14) bpm, P < 0.05]. No cardiac arrhythmia and hypotension were found in all neonates. (2) There were no significant differences on the ejection fraction, stroke volume and cardiac output of left ventricle between the two groups (P > 0.05). No significant difference was found in the numbers of left and right ventricular diastolic dysfunction, pulmonary hypertension between the two groups (P > 0.05). (3) The level of cardiac troponin T (cTnT) in plasma was (0.47 +/- 0.15) ng/ml in HG, and (0.35 +/- 0.21) ng/ml in NG, and there was no significant difference between the two groups (P > 0.05). CONCLUSION: No significant cardiac dysfunction complication caused by the hypothermia treatment was found in term neonates after birth asphyxia. PMID- 14749009 TI - [Value of cerebrospinal fluid leukocyte aggregation score in distinguishing the causes of meningitis]. PMID- 14749010 TI - [Relationship between oral and gastric Hp infection and diseases of upper digestive tract in children]. PMID- 14749011 TI - [Bacterial lipopolysaccharide sensitizes the immature brain to hypoxic-ischemic injury]. PMID- 14749012 TI - [Clinico-epidemiological analysis of cerebral palsy complicated diseases in children]. PMID- 14749014 TI - [A clinical study on spontaneous intracranial hemorrhage in children]. PMID- 14749013 TI - [Study of the relations between toxoplamosis and bronchial asthma]. PMID- 14749015 TI - [Congenital ectodermal displasia in a case]. PMID- 14749017 TI - [Analysis on 18 cases with necrotizing hyperplastic lymphadenopathy]. PMID- 14749016 TI - [Clinical observation of variable-flow nasal continuous positive airway pressure in preterm neonates with respiratory failure]. PMID- 14749018 TI - [Case No. 110--Dyspnea with exertion for seven years]. PMID- 14749019 TI - [One case with first seizure showing status epilepticus]. PMID- 14749020 TI - [Report of a case with protein-losing gastroenteropathy]. PMID- 14749021 TI - North American Contact Dermatitis Group patch-test results, 1998 to 2000. AB - BACKGROUND: Patch testing is the most worthwhile diagnostic tool for the evaluation of patients with suspected allergic contact dermatitis. OBJECTIVE: This study reports patch-testing results from July 1, 1998, to December 31, 2000, by the North American Contact Dermatitis Group. METHODS: Patients were tested with the same screening series of allergens, using a standardized patch-testing technique. The data from these patients were recorded on a standard computer entry form and analyzed. RESULTS: Fifty allergens were tested on over 5,800 patients. Amidoamine, benzophenone-3, and iodopropynyl butylcarbamate were the new allergens. The top 10 allergens in frequency of positive reactions were identical to those of our 1996-to 1998-study period. The incidence of allergic nickel reactions continues to go up, leading all the test substances by 16.2%. CONCLUSION: Our findings reinforce the need for a more comprehensive group of diagnostic allergens than is found in the T.R.U.E. TEST, which is sold in the United States. PMID- 14749022 TI - Atopy, nickel sensitivity, occupation, and clinical patterns in different types of hand dermatitis. AB - BACKGROUND: The etiologic diagnosis of hand dermatitis (HD) is often difficult. Knowledge of the relationship between atopy and nickel sensitivity as risk factors for HD is limited. OBJECTIVE: To compare irritant contact dermatitis, allergic contact dermatitis, and dyshidrotic eczema relative to personal atopy, patch-testing results, occupation, and clinical patterns. METHODS: From patients referred for patch testing, 714 consecutive individuals with HD were retrospectively studied. RESULTS: Half of the patients had eczema confined to the palms. The dorsal pattern was more prevalent in atopic patients, compared to nonatopic patients. Irritant contact dermatitis was the most frequent diagnosis (55.3%), followed by allergic contact dermatitis (24.4%) and dyshidrotic eczema (20.3%). Among housewives, health workers, and mechanics, irritants were of much greater importance than allergens whereas among hairdressers and bricklayers, the opposite was found. The prevalence of personal atopy did not differ significantly between different types of HD. Nickel sensitivity was much less frequent in irritant contact dermatitis than in allergic contact dermatitis or dyshidrotic eczema. The distribution of clinical patterns was similar. CONCLUSION: This study suggests that different etiologic diagnoses of HD cannot be distinguished by clinical pattern, prevalence of personal atopy, or nickel-sensitivity. PMID- 14749023 TI - Eyelid dermatitis: an evaluation of 447 patients. AB - BACKGROUND: Eyelids can be affected by various types of dermatitis that are often difficult to diagnose. OBJECTIVE: The aim of the study was to establish some guidelines for a correct diagnosis. METHODS: A total of 447 patients treated at 12 research units for eczema or other inflammatory dermatitis located on the eyelids were invited to complete a questionnaire. When necessary, patch tests with haptens of the standard series from Gruppo Italiano di Ricerca sulle Dermatiti da Contatto e Ambientali della Societa Italiana di Dermatologia e Venereologia (SIDEV-GIRDCA) were performed. RESULTS: Of the subjects studied, 50.2 % were diagnosed with allergic contact dermatitis (ACD); 20.9% were affected by irritant contact dermatitis (ICD), 13.5% by atopic dermatitis, 6.3% by seborrheic dermatitis, 6.5% by aspecific xerotic dermatitis, and 2.3% by psoriasis. Approximately 91% of all subjects reported an absence of familial atopy. A significant statistical association between diagnosis type and a personal history of atopy was evident (p <.000001, chi-square test). The results of gradual logistic regression models showed four-eyelid involvement as the main risk factor for ACD (odds ratio [OR] = 3.0; 95% CI, 1.1-8.1); with ICD, the main risk factor was the onset of symptoms at between 2 and 6 months (OR = 2.1; 95% CI, 1.1-4.0), whereas for atopic dermatitis, the main risk factors were the onset of symptoms later than 6 months and a personal history of atopy (OR = 4.9 and 3.6, respectively). CONCLUSION: Results suggest that many characteristics of the patients examined can be used for the differential diagnosis of palpebral eczematous dermatitis. PMID- 14749024 TI - Allergic contact cheilitis in the United Kingdom: a retrospective study. AB - BACKGROUND: To date, only a few cohorts of patients with allergic cheilitis have been described, most of them from Australia and Asia. OBJECTIVE: To establish the prevalence of cheilitis in a UK specialist contact dermatitis clinic and to identify the most common allergens. METHOD: We analyzed our patch-test database in a tertiary referral center in the United Kingdom, retrospectively. All patients presenting with cheilitis over a 19-year period (1982 to 2001) were included. RESULTS: Data were available from a total of 146 patients. A positive allergic patch-test reaction was thought to be relevant in 15% of the patients (n = 22) and to be of possible relevance in 6.8% (n = 10). Of the 22 patients with relevant allergic results, 95% (n = 21) were women. The most common allergens included fragrance mix (mainly cinnamaldehyde, oak moss, and isoeugenol) in 41% of patients, shellac in 18%, colophony in 18%, and Myroxylon pereirae in 14%. For half of the patients, the allergen was believed to stem from lipsticks or lip products. Eighteen percent of patients with allergic cheilitis reacted to only their own products. CONCLUSIONS: Patients should be tested to extended lipstick/cosmetic vehicle series in addition to standard series. As a significant percentage of patients react to their own products only, a thorough clinical history and testing to patients' own products are important. PMID- 14749025 TI - Sensitization to palladium chloride: a 10-year evaluation. AB - BACKGROUND: Palladium is increasingly used in industry, jewelry, and dentistry and is becoming more common since the European directive restricting the use of nickel in all products placed in direct and prolonged contact with the skin. OBJECTIVE: The role of palladium sensitization is still unclear, and the aim of our study is to evaluate the trend of sensitization in a contact dermatitis clinic population during a 10-year period. METHODS: We report our experience with 4,446 patients (3,077 female, 1,369 male; mean age, 40.1 +/-13.7 years) with suspected contact dermatitis patch-tested during the period of 1991 to 2000. RESULTS: A positive patch-test result to palladium chloride 1% was indicated in 236 patients (5.3%), with a higher percentage in females (6.7%) than in males (2.3%), and the sensitization to this metal has increased over the specified period, to a maximum in the year 2000 (9.7%). In the majority of cases, subjects were polysensitized (92.8%), but 7.2% of subjects were patch-test positive only to palladium. Of palladium-sensitized patients, 40.5% complained of hand dermatitis, 47.4% complained of body dermatitis, and 1.7% complained of burning mouth syndrome. Palladium sensitization is significantly related to female sex (OR = 3.08; 95% CI, 2.07-4.61) and to sensitization to other metals, with a maximum for nickel sulfate (OR = 32.9; 95% CI, 21.3-51.5). CONCLUSION: Sensitization to palladium is increasing by the year, reaching high values, but its role in reducing sensitization and symptoms is still unclear because of the frequent cosensitization with nickel and the low number of monosensitized patients with relevant sensitizations. We need to follow palladium sensitization in future years to verify an increase of sensitization due to the increasing use of this metal. PMID- 14749026 TI - Structure-activity relationships in allergic contact dermatitis. Part III. The sensitizing capacity of substituted phenanthrenequinones: a quantum-mechanical approach. AB - BACKGROUND: Nonterpenoid and diterpenoid phenanthrenequinones (PACs) have been found in the plant kingdom. Some of them occur in plants used in traditional Chinese medicine like Tan-Shen whereas others are constituents of orchids that are popular as ornamental plants. OBJECTIVE: Case reports and our own observations in orchid nurseries suggest that some or even all of these PACs possess a distinct sensitizing potency. Occasional exposure (particularly of botanists) to field-grown orchids, as well as occupational contact with sawdust of PAC-containing tropical timbers, caused allergic contact dermatitis. However, experimental studies in guinea pigs to determine the sensitizing capacity of PACs have not been performed so far. METHODS: Guinea pigs were sensitizied by a modified Freund's complete adjuvant method with four naturally occurring and 22 synthetic PACs in order to find out which and how many substituents at the carbons of the three rings of the PAC will influence the sensitizing power of the molecule. Subsequently, the lowest unoccupied molecular orbital (LUMO) coefficients were calculated to show whether a correlation exists between chemical reactivity and sensitizing capacity. RESULTS: Sensitizing capacity was found to be strong in two PACs, moderate in eight PACs, and weak in ten PACs. Five PACs were extremely weak in sensitizing capacity, and one PAC was completely negative. Two substituents on the left-hand carbons C-7 and C-8 of ring C were shown to be responsible for a strong sensitizing capacity. One methoxy group alone or three of them, especially when localized at C-5, decreased the sensitizing capacity to moderate. Substitution with a methoxy group at C-3 and/or at C-2 of the quinonoid ring itself (ring A) led to a weak sensitizing capacity. The ortho-quinones 1,2-PAC and 9,10-PAC were also weakly sensitizing. In fact, LUMO coefficient calculations corroborated a good correlation between chemical reactivity and sensitizing capacity. CONCLUSION: Substitution with methoxy groups at C-7 and/or at C-8 of ring C of 1,4-phenanthrenequinone increases the LUMO coefficients at the 2,3 double bond of ring A and thus facilitates nucleophilic substitution of protein nitrogen or sulfur nucleophiles at this electron deficient double bond. The four naturally occurring PACs that were investigated- cypripedin, denbinobin, annoquinone-A, and latinone--do not fulfill these criteria and are thus only weak sensitizers. However, as-yet-unstudied phenanthrenequinones occurring in plants or trees and having no substituents at C 2 or C-3 of the quinonoid ring must be considered potentially strong allergens. PMID- 14749027 TI - Generalized eczematous contact dermatitis from cocobolo wood. AB - Occupational contact with cocobolo wood (Papilionaceae, Dalbergia retusa) has been reported to rarely cause delayed hypersensitivity reactions. We report the case of a 53-year-old furniture and cabinetmaker who exhibited a generalized reaction mimicking erythroderma after exposure to sawdust from the wood. Patch testing to plants and woods standard (Chemotechnique, Dormer Laboratories, Ontario, Canada) was negative, and the specific allergen in cocobolo, obtusaquinone, was not available to us. The patient was tested instead to shavings of various woods as well as to sawdust of the suspected wood in petrolatum. He exhibited an exuberant response (+++) to both shavings and sawdust of cocobolo. After successful patch testing with shavings and sawdust in the absence of the purified chemical allergen, avoidance of the timber resulted in the resolution of his symptoms. PMID- 14749028 TI - Dermatitis artefacta? AB - A 35-year-old man presented with a 2-month history of intensely pruritic excoriated and crusted linear lesions on the dorsa of the left hand and left forearm (Fig 1). The patient had worked in construction for 2 years, and his job consisted mainly in covering the facades of buildings with cement. The patient was right-handed and used a black rubber glove as a protective measure only on his left hand (Fig 2). He reported that the lesions resolved partially during holidays and weekends and clearly flared in association with his work. There was no history of atopic dermatitis, drug use, or intolerance to metals, rubber, or fruits. On physical examination, linear excoriations with crusts were observed on the dorsa of the left hand, extending to the ventral and dorsal aspects of the forearm, involving the whole area that was in contact with the glove. Lichenified erythematous plaques and excoriations on the dorsal surface of the metacarpophalangeal joints and scaly lesions on the dorsal surfaces of the fingers were also present. On the palm, only discrete hyperkeratosis was seen. The right hand and forearm were free of lesions. He complained of intense pruritus when wearing the rubber glove and admitted to continuous scratching to relieve his discomfort, inducing the linear and excoriated lesions. Treatment with topical corticosteroids was initiated, with progressive resolution of the lesions. PMID- 14749029 TI - Common shoe allergens undetected by commercial patch-testing kits: dithiodimorpholine and isocyanates. AB - The diagnosis of allergic contact dermatitis of the feet is challenging because the constituents of shoes are not labeled. In addition, the materials and technologies used in the manufacturing of shoes are continuously evolving. Since the 1950s, rubber allergens have been the most common cause of shoe dermatitis. However, the causal allergens in rubber have changed. Instead of 2 mercaptobenzothiazole, dithiodimorpholine, an allergen that is not contained on standard patch-testing trays, now may be one of the more common rubber sensitizers in shoes. Other allergens not found on standard series that can account for shoe dermatitis include isophorone diisocyanate, dimethylaminoethyl ether, and mixed dialkyl thioureas. Therefore, physicians evaluating patients with allergic-appearing foot dermatitis need to test with allergens beyond those present on standard screening series. PMID- 14749030 TI - Barrier creams: fact or fiction. PMID- 14749031 TI - Considerations for testing irritancy, allergy, and photoreactivity in fragrance safety evaluations. AB - The aims of the Research Institute for Fragrance Materials, Inc. (RIFM), an international nonprofit science-based organization established in 1966 by the fragrance industry, include the prevention of adverse cutaneous effects, systemic toxicity, and environmental consequences from fragrance ingredients. This paper gives an overview of the RIFM testing and research program, how priorities are established, and how RIFM's safety evaluation process works. PMID- 14749033 TI - ACDS President's message. An important period in the history of our Society. PMID- 14749032 TI - Label dermatitis. PMID- 14749034 TI - Increased hypocretin-1 (orexin-a) levels in cerebrospinal fluid of rats after short-term forced activity. AB - The hypocretins (orexins) are recently discovered neuropeptides initially associated with feeding behavior and sleep regulation. However, the normal function of these peptides is unclear and a number of studies have reported a role in energy homeostasis and locomotor activity. Exercise (or physical activity) is the most powerful way of challenging the internal homeostatic process. This study examines the circadian differences in response to forced activity and homeostatic challenges on hypocretin-1 (Hcrt-1) levels in the cerebrospinal fluid (CSF) of rats. Hcrt-1 levels were decreased after long-term immobilization at the end of active phase (zeigeber time-0, ZT-0) and increased after short-term forced swimming in the rest phase (ZT-8). Nevertheless, no effects were observed after short-term immobilization, total sleep deprivation or cold exposure. We concluded that despite the relation between hypocretins, stress and sleep regulation reported in the literature, short-term total sleep deprivation, immobilization and cold exposure did not induce increases in CSF Hcrt-1 levels at ZT-0 and ZT-8. On the other hand, the relationship between hypocretinergic system activation and motor activation is reinforced by decrease in Hcr-1 levels after long-term immobilization at ZT-0 and its increased levels after short-term forced swimming at ZT-8 in CSF of rats. PMID- 14749035 TI - Amylin-induced suppression of ANP secretion through receptors for CGRP1 and salmon calcitonin. AB - Amylin cosecretes with insulin from pancreatic beta-cells and shows high sequence homology with CGRP, adrenomedullin, and salmon calcitonin. This study aimed to investigate the effect of amylin on the atrial hemodynamics and ANP release from rat atria and to identify its receptor subtypes. Isolated perfused left atria from either control or streptozotocin-treated rats were paced at 1.3 Hz. The concentration of ANP was measured by radioimmunoassay and the translocation of ECF was measured by [3H]-inulin clearance. Rat amylin increased atrial contractility and suppressed the release of ANP. Rat CGRP showed similar effects but was approximately 300-fold more potent than amylin. Pretreatment with receptor antagonist for CGRP1 [rat alpha-CGRP (8-37)] or salmon calcitonin [acetyl-(Asn30, Tyr32)-calcitonin(8-32), (AC 187)] blocked the suppressive effect of ANP release and the positive inotropic effect by rat amylin. However, receptor antagonists for amylin [amylin (8-37), acetyl-amylin] did not block those effects. Amylin (8-37), acetyl-amylin, or rat alpha-CGRP (8-37) alone accentuated the release of ANP with no changes in atrial contractility. The effect of rat amylin and rat amylin (8-37) on the ANP release was attenuated in streptozotocin treated rats. We suggest that amylin suppressed ANP release with increased atrial contractility through receptors for CGRP1 and salmon calcitonin and the attenuation of amylin and its antagonist on ANP release from streptozotocin treated rat atria may be due to the downregulation of amylin receptor. PMID- 14749036 TI - Functional significance of gastrin gene expression in human cancer cells. AB - The gastrointestinal peptide, gastrin, stimulates the growth of human pancreatic cancer. A receptor for gastrin activity, the cholecystokinin-C (CCK-C) receptor, has been identified in binding assays, cloned and sequenced, and is a splice variant of the CCK-B receptor. The relationship of gastrin and the CCK-C receptor to the growth of cancer cells was examined in vitro and in vivo. Stable transfection of the sense cDNA of gastrin into human MDA Amp-7 ampullary cancer cells, which normally lack gastrin gene expression but possess CCK-C receptors, increased cell growth up to 10-fold over wild type (WT) and vector-transfected (VT) cells. MDA Amp-7 tumors of gastrin-transfected cells reduced latency time for a visible tumor by 35%, decreased the timetable of tumor incidence, and increased tumor size by at least 2-fold in comparison to WT and VT groups. Transfection of human BxPC-3 pancreatic cancer cells, which normally express gastrin and possess CCK-C receptors, with the antisense cDNA to human gastrin decreased cell number by 30% in culture and tumor size by 53% compared to the WT and VT groups. Transfection of sense gastrin cDNA to monkey COS-1 cells, which normally lack both the gastrin and the CCK-C receptor genes, had no effect on growth. These studies demonstrate that gastrin and the CCK-C receptor form an autocrine loop in human pancreatic cancer that plays a role in regulating growth. PMID- 14749037 TI - Capsaicin-sensitive neural pathway mediates atrial natriuretic factor (ANF) release in response to physiological stimuli. AB - Increases in intravascular volume are detected by mechanoreceptors situated at the junctions of the great veins with the atria. We had previously shown that localized distension of the superior vena caval/right atrial junction, simulating increased cardiac preload, elicits release of ANF remotely from the atrial appendage. We proposed that ANF secretion is stimulated via intrinsic neural pathways running from the venoatrial junctions to the appendage. We developed a technique whereby non-adrenergic, non-cholinergic sensory nerves could be selectively destroyed in the heart of adult rats by instilling capsaicin into the pericardial space. Four days later, the animals were killed, and isolated perfused atria were prepared with small balloons positioned so that the superior vena caval/right atrial junction could be discretely stretched. Immunoreactive ANF secretion into the perfusate was measured. Although distension of the venoatrial junction increased ANF secretion from the control atria, there was no such response in the denervated atria. We conclude (A) that local application of capsaicin to the heart of adult rats induces selective functional neural deficits and (B) that information regarding distension of the junction of the great veins and the atria is normally transmitted across the atrium via these nerves to stimulate ANF secretion from peptide stores located in the atrial appendage. We propose that these pathways are crucial to ensure appropriate ANF secretion in response to an increase in circulating blood volume. PMID- 14749038 TI - Endothelin causes contraction of human esophageal muscularis mucosae through interaction with both ETA and ETB receptors. AB - Endothelin (ET) causes contraction of the muscularis mucosae in the guinea pig esophagus, but its role in the human esophagus remains unknown. To investigate effects of ET in the human esophagus, we measured contraction of isolated human esophageal muscularis mucosae strips caused by ET related peptides and binding of 125I-ET-1 to cell membranes prepared from the human esophageal muscularis mucosae. Autoradiography demonstrated specific binding of 125I-ET-1 to the muscularis mucosae and muscularis propria (muscularis externa) of the human esophagus. ET-1 caused tetrodotoxin and atropine-insensitive contraction of muscularis mucosae strips. In terms of the maximal tension of contraction, ET-1 and ET-2 were equal in efficacy. The relative potencies for ET related peptides to cause contraction were ET-1=ET-2>ET-3>sarafotoxin S6c (SX6c), an ETB receptor agonist. ET-1 caused contraction was mildly inhibited by BQ-123, an ETA receptor antagonist, and not by BQ-788, an ETB receptor antagonist. It was moderately inhibited by the combination of both antagonists, indicating synergistic inhibition. Furthermore, desensitization to SX6c with SX6c pretreatment failed to abolish the contractile response to ET-1, which was completely inhibited by BQ 123. These indicate the involvement of both ETA and ETB receptors in the contraction. Binding of 125I-ET-1 to cell membranes of the muscularis mucosae was saturable and specific. Analysis of dose-inhibition curves demonstrated the presence of ETA and ETB receptors. This study demonstrates that, the muscularis mucosae of the human esophagus, similar to that of the guinea pig esophagus, possesses both ETA and ETB receptors mediating muscle contraction. PMID- 14749039 TI - Insulin stimulates placental leucine aminopeptidase/oxytocinase/insulin-regulated membrane aminopeptidase expression in BeWo choriocarcinoma cells. AB - Placental leucine aminopeptidase (P-LAP), a cystine aminopeptidase that is identical to insulin-regulated membrane aminopeptidase, hydrolyzes oxytocin, which results in the loss of oxytocin activity. We previously isolated genomic clones containing the human P-LAP promoter region, which included two sites homologous to the 10-bp-insulin responsive element (IRE) that was identified on the phosphoenolpyruvate carboxinase gene. We therefore postulated that insulin regulates P-LAP expression via these IREs and investigated this notion using BeWo choriocarcinoma trophoblastic cells cultured in the presence of insulin. Insulin increased P-LAP activity in a time- and dose-dependent manner. Physiological concentrations of insulin at 10(-7) M exhibited the most potent effect on P-LAP activity. Western blotting demonstrated that 10(-7) M insulin increased P-LAP protein levels. Semi-quantitative RT-PCR and Southern blotting showed that insulin also increased P-LAP mRNA, which was abrogated by prior exposure to cycloheximide. Luciferase assay did not reveal any regulatory regions within 1.1 kb upstream of the P-LAP gene that could explain the insulin-induced P-LAP mRNA accumulation. These findings indicate that insulin induces P-LAP expression in trophoblasts, and that it acts via de novo synthesis of other proteins, which partially contradicts our initial hypothesis. PMID- 14749040 TI - Immunohistochemical colocalization of type II angiotensin receptors with somatostatin in rat pancreas. AB - Earlier studies indicate that binding sites of type II angiotensin (AT2) receptors are detected all over the pancreas, as well as in the pancreatic exocrine cell line AR4-2J. However, lack of corresponding functional AT2 receptor responses can be detected in the exocrine pancreas. The aim of present study is to determine the protein expression of AT2 receptors in the pancreas by probing with an AT2 receptor-specific antibody, and to examine the role of AT2 receptors in the regulation of pancreatic endocrine hormone release. In Western protein analysis of adult rat tissues, expression of AT2 receptor-immunoreactive bands of 56, 68, and 78 kDa was detected in the adrenal, kidney, liver, salivary glands, and pancreas. In adult rat pancreas, strong immunoreactivity was detected on cells that were located at the outer region of Langerhans islets. Immunohistochemical studies indicated that AT2 receptors colocalized with somatostatin-producing cells in the endocrine pancreas. Consistent with the findings in adult pancreas, abundant expression of AT2 receptors was also detected in immortalized rat pancreatic endocrinal cells lines RIN-m and RIN-14B. To examine the role of AT2 receptors on somatostatin secretion in the pancreas, angiotensin-stimulated somatostatin release from pancreatic RIN-14B cells was studied by an enzyme immunoassay in the absence or presence of various subtype selective angiotensin analogues. There was a basal release of somatostatin from RIN-14B cells at a rate of 8.72 +/- 4.21 ng/10(6) cells (n = 7). Angiotensin II (1 nM-10 microM) stimulated a biphasic somatostatin release in a dose-dependent manner with an apparent EC50 value of 49.3 +/- 25.9 nM (n = 5), and reached maximal release at 1 microM angiotensin II (982 +/- 147.34% over basal secretion; n = 5). Moreover, the AT2 receptor-selective angiotensin analogue, CGP42112, was 1000 times more potent than the AT1 receptor-selective angiotensin analogue, losartan, in inhibiting angiotensin II-stimulated somatostatin release. These results suggest that angiotensin may modulate pancreatic hormone release via regulation of somatostatin secretion. PMID- 14749041 TI - Effects of des-aspartate-angiotensin I on the expression of angiotensin AT1 and AT2 receptors in ventricles of hypertrophic rat hearts. AB - The effects of des-aspartate-angiotensin I (DAA-I) on the expression of angiotensin AT1 and AT2 receptor in hearts of aortic coarcted rats were studied. The protocols used included competitive reverse transcription polymerase chain reaction (RT-PCR), Western blotting, and receptor-ligand binding assays. mRNA of the AT1 and AT2 receptors increased significantly after 4 days of aortic coarctation (7- and 4-folds of sham-operated, respectively). However, the protein of the AT1 receptor was not altered, and only increase in protein of the AT2 receptor was detected. There was an increase in [125I]Sar1-Ile8-angiotensin II binding sites in the ventricular membranes of hypertrophic hearts, which was attributed to an upregulation of the AT2 receptor. Treatment with i.p. DAA-I resulted in a significant reduction of cardiac hypertrophy, the maximum effect was achieved with a dose of 200 nmol/kg/day. The anti-cardiac hypertrophy effect appeared to be U-shape, and at a higher dose of 800 mol/kg/day, there was a loss of effect. DAA-I had no effect on the receptor protein in ventricles of hypertrophic hearts. However, DAA-I dose-dependently decreased the binding of [125I]Sar1-Ile8-angiotensin II to ventricular membranes. The decrease was due to a likely desensitization by internalization of the AT1 receptor, and this probably contributed to the loss of hypertrophic effects at 800 nmol/kg/day. Treatment of DAA-I also resulted in a remarkable increase in AT2 receptor mRNA (24-fold increase over the sham-operated), which was not coupled to translation. The present findings provide new information regarding the relationship between cardiac hypertrophy and the angiotensin receptors, and the anti-cardiac hypertrophic actions of DAA-I via the AT1 receptors. PMID- 14749042 TI - Effects of des-aspartate-angiotensin I on neointima growth and cardiovascular hypertrophy. AB - The in vitro anti-hypertrophic and hyperplastic actions of des-aspartate angiotensin I (DAA-I) on cultured cardiovascular cells have been demonstrated in earlier experiments. The present study investigated its effects on the development of neointima in balloon catheter-injured carotid artery of the Sprague-Dawley (SD) rat and the development of cardiovascular hypertrophy in the spontaneously hypertensive rat. Treatment with i.v. DAA-I for 14 days post-injury dose-dependently attenuated the development of neointima. The maximum effect was obtained at 34 pmol/kg/day. The data support the possibility that endogenous angiotensins could inhibit neointima growth. This opens up avenues for their therapeutic elevation in combating neointima-related restenosis of which current drugs are not fully effective in suppressing. Five-week-old pre-hypertensive SHR, when orally administered with a dose of 769 nmol/kg/day DAA-I for a duration of 47 weeks, showed significant reduction in the development of cardiac and vascular hypertrophy compared to the untreated controls. Similar treatment with DAA-I had no effect on the Wistar Kyoto rats. The present findings support the contention that, besides angiotensin II, other endogenous angiotensins are also involved in the regulation and/or pathophysiology of the cardiovascular system. PMID- 14749043 TI - A panel of 11 region-specific radioimmunoassays for measurements of human chromogranin A. AB - INTRODUCTION: The primary structure of human chromogranin A (CgA) not only contains 10 pairs of basic amino acids, which are potential cleavage sites for specific endogenous proteases, but also other sites in the molecule can be subjected to cleavage. Several CgA-related peptides have been identified in tissue, and many of the biological effects attributed to CgA seem to be mediated by these peptides. MATERIALS AND METHODS: Peptides homologous to defined parts of the human CgA molecule were selected and synthesised. Antibodies were raised, and 11 specific radioimmunoassays were developed. Plasma samples from 20 patients with neuroendocrine tumours were collected and measured in all assays. RESULTS: All assays measured circulating levels of CgA-derived peptides. Only four of the assays measured concentrations that correlated with that of total CgA. However, concentrations of the individual CgA-related peptides were generally lower than the concentration of total CgA. Different neuroendocrine tumours seem to process CgA differently. The ratio between a given region-specific assay and total CgA is inversely correlated to tumour activity. CONCLUSION: The assays presented allow measurements of defined regions of CgA and will thus become important tools for further studies of processing of CgA. PMID- 14749044 TI - Use of Ovsynch in dairy herds--differences between primiparous and multiparous cows. AB - Ovsynch protocols are used to increase service rate and decrease days open and cullings for infertility. Recent reports have indicated better results after Ovsynch in primiparous than in older cows. However, this was not observed in all investigations on the subject. The objective of the study was to evaluate differences between primiparous and multiparous cows after synchronization of ovulation with an Ovsynch protocol in six trials. A total of 1584 cows (583 primiparous and 1001 multiparous cows, respectively) on three dairy farms were synchronized with an Ovsynch protocol consisting of a GnRH-analogue at Days 0 and 9, and a prostaglandin F(2alpha) analogue on Day 7. AI was carried out in all cows 16-20 h after the last treatment. Cows were categorized into primiparous and multiparous cows for analysis. Conception rate (CR) to timed AI, to further AI, overall conception rate and proportion of cows pregnant by 200 days in milk were compared between the age groups. Finally, two logistic regression models were calculated with conception to first service and conception by 200 DIM as the outcome variables. Independent variables were trial (categorical) and age group (primiparous versus multiparous). Conception rates to TAI were higher in primiparous than in older cows (37.9% versus 31.6%, P=0.015). Likewise pregnancy rates by 200 DIM were higher in primiparous cows (81.8% versus 75.4%, P=0.003). However, the extent of the difference varied between trials. Results indicate that Ovsynch protocols are more effective in primiparous than in older cows. PMID- 14749046 TI - Fertility in beef cattle given a new or previously used CIDR insert and estradiol, with or without progesterone. AB - The objective was to compare pregnancy rates following fixed-time AI (FTAI) in beef cattle given a new or previously used CIDR insert and injections of estradiol, with or without progesterone, to synchronize follicular wave emergence. In Experiment 1, heifers (n=616) received a new or once-used CIDR insert for 9 days and were given 1mg estradiol cypionate (ECP), with or without 100 mg of a commercial progesterone preparation (CP4), at CIDR insertion. Heifers were treated with PGF at CIDR removal and 0.5 mg ECP i.m. 24h later, with FTAI 55 to 60 h after CIDR removal. Pregnancy rate was not affected by either the number of CIDR uses (P=0.59; 48.3% versus 46.2% for new versus once-used CIDRs, respectively) or the addition of progesterone (P=0.42; 45.6% versus 48.8% for ECP+CP4 and ECP, respectively). In Experiment 2 (replicated at two locations), heifers (n=56) and lactating beef cows (n=307) received a once- or twice-used CIDR and an i.m. injection of 1mg estradiol benzoate (EB), with or without 100 mg progesterone, at CIDR insertion. Cattle received PGF in the ischiorectal fossa at CIDR removal (Day 7) and 1mg EB i.m. 24h later, with FTAI 52 to 56 h after CIDR removal. Pregnancy rate was affected by location (P<0.002; 46.0% versus 61.1% for Locations A and B, respectively), parity (P<0.04; 67.9% versus 53.1% in heifers and cows, respectively), and numbers of times the CIDR had been used (P<0.03; 62.4% versus 48.4% for once- and twice-used CIDRs, respectively). However, the addition of progesterone to the injection of EB at CIDR insertion did not affect pregnancy rate (P=0.6). In Experiment 3, heifers (n=187) received one new, one once-used, one twice-used or two twice-used CIDRs for 7 days and 2 mg EB plus 50 mg of CP4 at the time of CIDR insertion. Heifers were treated with PGF at CIDR removal and 1mg EB i.m. 24 h later, with FTAI 52-56 h after CIDR removal. Pregnancy rate was not affected by treatments (P=0.28, 57.5, 63.8, 47.9, 47.9% for one new, one once-used, one twice-used, or two twice-used CIDRs, respectively). In summary, pregnancy rate to FTAI did not differ between cattle synchronized with a new or once-used CIDR, but pregnancy rate was lower in cattle synchronized with a twice-used CIDR; however, the insertion of two twice-used CIDRs did not affect pregnancy rates. The addition of an injection of progesterone to the estradiol treatment at CIDR insertion did not enhance pregnancy rate to FTAI. PMID- 14749045 TI - Effect of intrauterine administration of oestradiol on postpartum uterine bacterial infection in cattle. AB - After parturition fewer first dominant follicles are selected in the ovary ipsilateral to the previously gravid uterine horn in cattle. However, the presence of a large oestradiol-secreting follicle in the ipsilateral ovary is a predictor of fertility, possibly due to a localised effect of oestradiol which increases the rate of elimination of the ubiquitous uterine bacterial contamination that occurs after calving. The present study tested the hypothesis that oestradiol reduces uterine bacterial contamination when administered into the uterine lumen around the expected time for selection of the first postpartum dominant follicle. Animals were infused with saline (n=15) or 10mg oestradiol benzoate (n=15) into the previously gravid uterine horn on Days 7 and 10 postpartum. Peripheral coccygeal blood samples were collected daily and oestradiol concentrations measured by radioimmunoassay (RIA). Uterine lumen swabs were collected 7, 14 and 21 days postpartum for aerobic and anaerobic culture, bacteria were identified and growth scored semi-quantitatively. Plasma oestradiol concentrations were higher for treated animals between Days 7 and 14 (1.4+/-0.1 versus 2.0+/-0.2 pg/ml, P<0.05). Control animals had a similar bacterial growth score on Days 7 and 14, with a lower value on Day 21 (5.7+/-1.0 and 6.1+/-0.7 versus 0.3+/-0.1, P<0.05). However, treated animals had a surprising higher bacterial load on Day 14, than on Days 7 or 21 (7.1+/-0.9 versus 4.0+/-0.6 or 3.6+/-0.6, P<0.05). The increased score was attributable to more pathogens associated with endometritis on Day 14 than Day 7 (5.1+/-1.0 versus 2.5+/-0.5, P<0.05), in particular Prevotella melaninogenicus (1.5+/-0.5 versus 0.7+/-0.2, P<0.05) and Fusobacterium necrophorum (1.5+/-0.4 versus 0.3+/-0.2, P<0.05). In conclusion, administration of oestradiol into the uterine lumen surprisingly increased uterine pathogenic anaerobic bacterial contamination. Thus, it is unlikely that increased fertility associated with a first dominant follicle in the ipsilateral ovary is a consequence of the elimination of bacterial contamination by ovarian oestradiol. PMID- 14749047 TI - Activation and early parthenogenesis of bovine oocytes treated with ethanol and strontium. AB - Efficient artificial activation is indispensable for the success of cloning programs. Strontium has been shown to effectively activate mouse oocytes for nuclear transfer procedures, however, there is limited information on its use for bovine oocytes. The present study had as objectives: (1). to assess the ability of strontium to induce activation and parthenogenetic development in bovine oocytes of different maturational ages in comparison with ethanol; and (2). to verify whether the combination of both treatments improves activation and parthenogenetic development rates. Bovine oocytes were in vitro matured for 24, 26, 28, and 30 h, and treated with ethanol (E, 7% for 5 min) or strontium chloride (S, 10mM SrCl(2) for 5h) alone or in combination: ethanol+strontium (ES) and strontium+ethanol (SE). Activated oocytes were cultured in vitro in synthetic oviductal fluid (SOF) medium and assessed for pronuclear formation (15-16 h), cleavage (46-48 h) and development to the blastocyst stage (D7). Treatment with ethanol and strontium promoted similar results regarding pronuclear formation (E, 20-66.7%; S, 26.7-53.3%; P>0.05) and cleavage (E, 12.8-40.6%; S, 16.1-41.9%; P>0.05), regardless of oocyte age. The actions of both strontium and ethanol were influenced by oocyte age: ethanol induced greater activation rates after 28 and 30 h of maturation (48.4 and 66.7% versus 20.0 and 23.3% for 24 and 26 h, respectively; P<0.05) and strontium after 30 h (53.3%) was superior to 24 and 26 h (26.7% for both). Blastocyst development rates were minimal in all treatments (0.0-6.3%; P>0.05), however, when the mean (+/-S.D.) cell number in blastocysts at the same maturational period was compared, strontium treatment was superior to ethanol for activation rates (82+/-5.7 and 89.5+/-7.8 versus 54 and 61, at 28 and 30 h, respectively). Improved results were obtained by combined treatments. The combination of ethanol and strontium resulted in similar pronuclear formation (ES, 36.7-83.9%; SE, 53.1-90.3%) and cleavage rates (ES, 31.3-81.3%; SE, 65.6 80.7%). Regarding embryo development, there was no difference (P>0.05) between treatments, and blastocysts were only obtained in treatment SE at 24 and 26 h (6.5% for both). It is concluded that, SrCl(2) induces activation and parthenogenetic development in bovine oocytes. PMID- 14749048 TI - Relationship between thirty post-thaw spermatozoal characteristics and the field fertility of 11 high-use Australian dairy AI sires. AB - This study determined the relationship between two measures of field fertility of 11 high-use Australian artificial insemination (AI) dairy bulls and thirty standard laboratory assessments of spermatozoal post-thaw viability. The two measures of field fertility used, conception rates (cCR) and non-return rates (cNRR), were both corrected for all major non-bull variables. Sperm viability assessments were conducted on semen collected within the same season as that used to derive the field fertility estimates. These assessments measured sperm concentration, motility, morphology and membrane integrity at thawing, after 2h incubation and after the swim-up sperm selection procedure. Derivations of these measures and in vitro embryo fertilizing and developmental capacity were also determined. The Genstat Statistical Package [Genstat 5 Release 4.2 Reference Manual, VSN International, Oxford, 2000] was used to conduct an analysis of variance on the viability parameters across semen straws and bulls, and to calculate the strength of correlation between each semen parameter, cNRR and cCR in a correlation matrix. Step forward multiple regression identified the combination of semen parameters that were most highly correlated with cCR and with cNRR. The sperm parameters identified as being most predictive of cCR were the percentage of morphologically normal sperm immediately post-thaw (zeroNorm), the number of morphologically normal sperm after the swim-up procedure (nSuNorm), and the rate of zygote cleavage in vitro (Clv); the predictive equation formed by these parameters accounted for 70% of variance. The predictive equation produced for cNRR contained the variables zeroNorm, the proportion of membrane intact sperm after 2h incubation at 37 degrees C (twoMem) and Clv and accounted for 76.5% of the variation. ZeroNorm was found to be consistent across straws and semen batches within-bull and the sperm parameter with the strongest individual predictive capacity for both cCR (P=0.1) and cNRR (P=0.001). Post-thaw sperm parameters can be used to predict field fertility of Australian dairy sires; the calculated predictive equations are particularly useful for identifying and monitoring bulls of very high and very low potential fertility within a group. PMID- 14749049 TI - Effects of medroxyprogesterone acetate (MAP) on ovarian antral follicle development, gonadotrophin secretion and response to ovulation induction with gonadotrophin-releasing hormone (GnRH) in seasonally anoestrous ewes. AB - When ovulation is induced with gonadotrophin-releasing hormone (GnRH) in anoestrous ewes, a proportion of animals fail to form normal (full-lifespan) corpora lutea (CL). Progesterone treatment before GnRH prevents luteal inadequacy. It remains uncertain whether a similar effect, achieved with medroxyprogesterone acetate (MAP) from intravaginal sponges, is mediated by influences on growing ovarian follicles and/or secretion of gonadotrophic hormones, before and after GnRH treatment. Two experiments were performed, on 13 and 11 anoestrous Western white-faced ewes, respectively. Seven and six ewes, respectively, received MAP-containing sponges (60 mg) for 14 days; the remaining ewes served as untreated controls. To test the effect of timing of GnRH administration after pre-treatment with MAP-releasing sponges, GnRH injections (250 ng every 2h for 24h followed by a bolus injection of 125 microg of GnRH i.v.) were given either immediately (Experiment 1) or 24h after sponge removal in the treated ewes (Experiment 2). Ovarian follicular dynamics (follicles reaching >or=5mm in size) and development of luteal structures were monitored using transrectal ultrasonography. In Experiment 1, the mean ovulation rate (0.7+/-0.3 and 1.0+/-0.4) and proportion of ovulating ewes (57 and 67%, respectively) did not vary (P>0.05) between MAP-treated and control ewes. Normal (full-lifespan) CL were detected in 29% of treated and 67% of control ewes (P>0.05). In Experiment 2, the mean ovulation rate (2.3+/-0.2 and 1.2+/-0.6; P<0.05) and percentage of ewes with normal (full-lifespan) CL (100 and 40%, respectively; P<0.10) were greater in the treated compared to control ewes. In Experiment 1, the mean peak concentration of the GnRH-induced LH surge was lower (P<0.05) in MAP-treated than in control ewes. There were no significant differences between MAP-treated and control ewes in the characteristics of follicular waves, mean daily serum FSH concentrations, and secretory parameters of LH/FSH, based on intensive blood sampling conducted 1 day before sponging and 1 day before sponge removal. It is concluded that treatment with MAP has no effect on the tonic secretion of LH/FSH or follicular wave development in anoestrous ewes. However, the GnRH-stimulated LH discharge was attenuated in the ewes that received MAP-impregnated sponges for 14 days and were treated with GnRH immediately after sponge withdrawal. Ovulatory response and CL formation were increased when GnRH was administered 24 h after sponge removal. PMID- 14749050 TI - Effect of recombinant human FSH and LH on in vitro maturation of sheep oocytes; embryo development and viability. AB - The objective of this work was to study the effect of a preparation of human recombinant gonadotrophins (r-FSH and r-LH) on the in vitro maturation (IVM) and development of sheep oocytes. In addition, the viability of fresh and vitrified blastocysts obtained after transfer was tested. Oocytes collected from slaughtered animals were divided into five different maturation groups. All groups were matured in a medium containing TCM199 with 4 mg/ml BSA, 100 microM cysteamine and 1 microg/ml estradiol-17beta. Each group was also treated with one of the following: 0.1 UI/ml r-FSH (r-FSH group), 0.1UI/ml r-LH (r-LH group), 0.1 UI/ml r-FSH and 0.1 UI/ml r-LH (r-FSH/r-LH group), 5 microg/ml FSH and 5 microg/ml LH hypophysial gonadotrophins (h-G group) as a control, or no gonadotrophins (no-G group). After in vitro fertilization with fresh ram semen, presumptive zygotes were cultured in vitro for 6-7 days and a total of 109 blastocysts were then transferred in pairs into synchronized ewes. To determine the viability of embryos after vitrification, 36 blastocysts from the r-FSH/r-LH group and 30 from the h-G group were vitrified in 10% ethylene glycol (EG) and 10% dimethylsulphoxide (DMSO) for 5min, followed by 20% EG, 20% DMSO and 0.5M Sucrose (S) for <45 s. They were loaded into open pulled straws (OPS) and plunged into LN(2). After warming, the blastocysts were transferred in pairs into synchronized ewes. The highest maturation rate was reached in the r-FSH/r-LH group (91.9%). However, no statistical difference was found when this group was compared with the h-G group (84.0%). Likewise, the cleavage rate of the r-FSH/r LH group (81.4%) was not significantly different from that of the h-G group (82.3%). The cleavage rates of all other groups, however, were significantly lower than the r-FSH/r-LH and h-G groups. The blastocyst rate was highest in the h-G group (53.6%), and it was statistically higher than in the r-FSH/r-LH group (41.5%). The blastocyst rate was very similar between groups r-FSH and r-FSH/r-LH (42.0 and 41.5%, respectively). The lowest lambing rate (31.8%) was in the no-G group. The highest lambing rate was achieved in the r-FSH/r-LH group (66.6%). The vitrified embryos of h-G and r-FSH/r-LH groups had a very similar lambing rate (16.6% and 19.4%). In conclusion, these data provide support for the hypothesis that sheep oocytes respond to human recombinant gonadotrophins used for in vitro embryo production. PMID- 14749052 TI - Variability in relationships between semen quality and estimates of in vivo and in vitro fertility in boars. AB - The present experiment was designed to characterize relationships between common semen quality and fertility estimates for three boars known to differ in farrowing rate, number of pigs born alive, and monospermic penetration rate. The approach chosen to accomplish this was to monitor semen quality from these boars and use their semen alternately for either artificial insemination or in vitro fertilization for 40 weeks. This strategy relied on the variability in semen quality parameters that normally occurs in an individual boar over time. When comparisons were made among boars, farrowing rates, numbers of pigs born alive, and monospermic penetration rates were significantly different, but progressive motility, normal head and tail morphology, and acrosome morphology were not. However, when comparisons were made among ejaculates within individual boars, there were significant effects of semen quality on both in vivo and in vitro fertility. For boar 3495, the proportion of spermatozoa exhibiting progressive motility and distribution of spermatozoa in a percoll gradient had a positive linear effect on number born alive and monospermic penetration rate, respectively. For boar 2901, quadratic equations best described changes in litter size as a function of progressive motility and normal acrosomes. In addition, monospermic penetration rate increased linearly as normal acrosomes and the proportion of spermatozoa recovered from a percoll gradient increased. For boar 4291, the relationship between progressive motility and number born alive and between normal acrosomes and number of pigs born alive were also quadratic. However, a significant linear relationship was present only between normal acrosomes and monospermic penetration rate. These results demonstrate that simply relying on the means of common semen quality estimates from some boars has limited value in terms of being used as a prospective indicator of their in vivo or in vitro fertility. In contrast, characterization of relationships between semen quality and fertility estimates is useful for estimating differences in the fertility of ejaculates from individual boars. However, both quantitative and qualitative differences in these relationships among boars are present and a given semen quality estimate that is a good predictor of in vivo or in vitro fertilization for one boar, may not be applicable for others. PMID- 14749051 TI - Secretion of estradiol-17beta by porcine mammary gland of ovariectomized steroid treated sows. AB - Although the mammary gland of many species secretes estradiol (E(2)), nothing is known of E(2) secretion in the porcine gland. The present study was designed to investigate whether porcine mammary gland was a source of E(2), and to test the influence of individual and combined effects of exogenous progesterone and estradiol benzoate (EB) on the secretion of E(2). Immature crossbred gilts were ovariectomized at 7 months of age followed by 4 weeks later by steroid hormone replacement therapy to produce estradiol and progesterone (P(4)) blood concentrations similar to those observed during a normal estrous cycle. Arterial and venous blood plasma (from carotid artery and anterior mammary vein, respectively) were sampled for 2h at 10 min intervals. Plasma concentrations of progesterone, androstenedione (A(4)), testosterone (T), estrone (E(1)) and estradiol were determined by RIA. In all gilts treated with progesterone alone or in combination with EB, concentrations of P(4), A(4) and E(1) in blood collected from venous outflow were lower compared to concentrations in arterial blood, whereas concentrations of E(2) were higher in blood plasma from the anterior mammary vein compared to plasma from the carotid artery. The results indicated that the porcine mammary gland secreted E(2). Increased concentrations of plasma E(2) collected only from P(4)-treated animals suggested that progesterone activated enzymes involved in steroidogenesis in porcine mammary gland, or those utilized in its metabolism. PMID- 14749053 TI - Ovarian cysts and their consequences on the reproductive performance of swine herds. AB - The aim of this work was to determine the incidence of ovarian cysts in the breeding herd and their consequences in the reproductive performance of the herd. Data from 1990 cyclic sows from two farms, with 0-12 parities, lactation length between 6 and 47 days and weaning-estrus interval between 0 and 32 days were evaluated by ultrasound examination for cyst incidence. As cyst was considered an anaechoic structure with smooth and thin walls with a diameter larger than 2 cm that remained visible for at least 5 days after estrus onset. Cyst incidence was found to be 2.4%. Sows with ovarian cysts have a greater return to estrus rate (34.0 x 7.7%, P<0.01), and cysts were associated with around 10% of regular and irregular return to estrus patterns on both farms. The adjusted farrowing rate (52.2 x 90.0%, P<0.01) and anestrual sows that were not pregnant (10.6 x 0.6%, P<0.01) were also influenced by the appearance of ovarian cysts, but they did not influence litter size (P>0.05). The incidence of cysts was not influenced by parity (P>0.05). Sows with shorter lactation had a greater incidence of cysts (P<0.05). Sows with a weaning-estrus interval shorter than 3 days had a greater incidence of ovarian cysts (P<0.05). The time of the year had no influence on the incidence of ovarian cysts (P<0.05). PMID- 14749054 TI - Action of protein kinase A regulators on secretory activity of porcine granulosa cells in vitro. AB - To understand the role of protein kinase A (PKA) in the control of ovarian secretory activity, we examined effects of stimulators (db-cAMP, 6-Phe-cAMP, Sp cDBIMPS) or inhibitors (Rp-cAMPS, KT5720) of PKA on the release of insulin-like growth factor I (IGF-I), progesterone (P) and estradiol (E) by cultured porcine granulosa cells using RIA. All the PKA stimulators db-cAMP (10-10000 ng/ml), 6 Phe-cAMP (10-10000 pmol) or Sp-cDBIMPS (1-10000 pmol) increased IGF-I almost at all doses tested. P release was stimulated by db-cAMP (at doses 100-10000 ng/ml), Sp-cDBIMPS (at 10-1000 pmol) and 6-Phe-cAMP (at 1000 and 10000 pmol). The release of E was stimulated by Sp-cDBIMPS (1-100 pmol), db-cAMP (1000 and 10000 ng/ml) and 6-Phe-cAMP (1000 and 10000 pmol). Since Sp-cDBIMPS, which activates preferentially PKA isozyme type II, showed stimulating effects at doses lower than those of 6-Phe-cAMP, a preferential activator of both, type I and II of PKA, it is assumed that PKA type II is more important for the control of ovarian steroidogenesis than type I. A PKA inhibitor Rp-cAMPS inhibited release of IGF-I (10000 pmol), P (1000 pmol) and E (1000 and 10000 pmol), whereas Rp-cAMPS, at doses higher than 1000 pmol, tended to reverse this inhibitory effect. Other PKA inhibitor KT5720 suppressed P (at 10-1000 ng/ml), but not IGF-I or E release.The stimulation of growth factor and sex steroid release by PKA activators, and suppression of the secretion some of these substances by PKA inhibitors may indicate the implication of PKA (probably site B) in up- and down-regulation of ovarian IGF-I and steroid release. PMID- 14749055 TI - Effect of oestrous cycle and early pregnancy on uterine production and expression of immune regulatory factors in gilts. AB - This study was undertaken to characterize uterine immune factors involved in the establishment of pregnancy in gilts. Thirty crossbred Yorkshire-Landrace gilts of similar age and weight were observed twice a day for oestrous behaviour with intact boars. On the day of first standing oestrus (Day 0) and 12h later, 15 gilts were inseminated with pooled semen from Duroc boars of proven fertility. Pregnant gilts were slaughtered either on Days 10, 15 or 25 of gestation (n=5 per day). The other 15 gilts were not inseminated and were slaughtered on either Days 0, 10 or 15 of the oestrous cycle (n=5 per day). Immediately after slaughter, endometrial tissue samples from the mesometrial side were removed for gene expression using RNase protection assay and in situ hybridization methodologies. The other uterine horn was flushed with 20 ml of PBS to collect the uterine fluid. In pregnant gilts, endometrial interleukin (IL)-6 mRNA expression was higher on Day 15 than on Days 10 and 25 (P<0.01 and P<0.1, respectively). On Day 15, IL-6 expression was also significantly higher (P<0.01) in pregnant gilts than in cyclic gilts. In both pregnant and cyclic gilts, transforming growth factor (TGF)-beta2 in uterine fluid was significantly higher (P<0.0001) on Day 15 than on Day 10. At the gene expression level, TGF-beta2 also increased between Days 10 and 15 in both cyclic and pregnant gilts but differences were not significant. On Day 15, concentrations of interferon-gamma and prostaglandin E(2) (PGE(2)) in uterine fluid were markedly higher (P<0.001) in pregnant gilts than in cyclic gilts, whereas the total amount of TGF-beta2 in uterine fluid and its endometrial expression were approximately 70% higher although this increase was not significant. Finally, tumour-necrosis factor-alpha and granulocyte macrophage/colony-stimulating factor mRNA expressions were undetectable in all endometrial samples. In conclusion, production and/or expression of uterine TGF beta2, IL-6 and PGE(2) increased during the embryonic attachment period and are coincidental with embryonic interferon-gamma production. PMID- 14749056 TI - The effect of level of feeding in early gestation on reproductive success in young rabbit does. AB - An experiment examined the effect of restricted feeding during early gestation on later feed intake and on kindling performance in young rabbit does. Nulliparous rabbit does (n=94) were inseminated at 14.5 weeks of age. During the first 10 days of gestation, does were fed individually either to appetite (AL) or 1.35 times maintenance requirement (R). After 10 days of gestation, all animals were fed to appetite. Does were weighed at insemination and after kindling. Feed intake was recorded during the first 10 days of gestation and weekly, thereafter. The number of does that kindled, number of live and stillborn kits and litter weight was recorded. The experiment ended after kindling. During feed restriction, AL does consumed 70 g/day more on an average than R does (209 and 139+/-4 g for AL and R, respectively; P<0.001). In the first and second week after feed restriction, compensatory feed intake occurred. Restrictive fed does ate more than AL does (+34+/-5 g/day from days 11 to 17 of gestation; P<0.001; +17+/-6g/day from days 18 to 24 of gestation; P<0.05). In the last week of gestation, feed intake of AL and R was comparable (89 and 100+/-5 g, respectively). At kindling, AL does had consumed 8+/-3g/day more feed over the total gestation period than R does (P<0.04). Feeding level during early gestation neither affected the kindling rate (83%), total litter size (7.9+/-0.4) nor the number of does with stillborn kits (10 versus 9 for AL and R, respectively). Regardless of treatment, in the last week of gestation (days 25-32) the number of does with stillbirth was lower and average birth weight was higher (P<0.01) in does eating more than the average daily feed intake compared to does eating below average. Based on the results of this study it was concluded that feed restriction for 10 days in early gestation does not affect kindling performance of young does, but feed intake in the last week of gestation affects kit survival and birth weight. PMID- 14749057 TI - The potential for gamete recovery from non-domestic canids and felids. AB - Species are becoming extinct at a rate 100 times the natural background rates. Considering all mammalian orders, 24% of all Carnivora species are threatened. The goal of carnivore conservation is to reverse the decline in populations and to secure remaining populations in ways that will assure enduring public support. In this context, biotechnology is a tool with tremendous potential for assisting the conservation of endangered canid and felid species. As the first step for biotechnology development is the gamete obtainment, this review will discuss the potential of gamete recovery from non-domestic canids and felids, based on learning how to apply these procedures in the domestic carnivores. Thus, electroejaculation and obtaining both epidydimal spermatozoon and spermatogonial germ cells are indicated as techniques for male gametes recovery. In the female gametes retrieval, different methods for oocyte recovery from both antral and preantral follicles, and the possibility for ovarian tissue transplantation are discussed. Furthermore, the study discusses the responsibilities involved in the use of assisted reproduction in endangered species conservation. PMID- 14749058 TI - Intra- and inter-species differences in persistent organic contaminants in the blubber of blue whales and humpback whales from the Gulf of St. Lawrence, Canada. AB - Biopsy samples of blubber from adult male and female blue whales, and from female and young-of-the-year humpback whales were collected during the summers of 1992 1999 in the Gulf of St. Lawrence, Canada. In blue whales, concentrations of 25 PCB congeners, DDT and metabolites and several other organochlorine compounds were present at higher concentrations in the blubber of males relative to females; reflecting maternal transfer of these persistent contaminants from females into young. Sex-related differences in concentrations were not observed with less persistent contaminants, such as HCHs. In humpback whale samples, there were no significant differences in the concentrations of PCBs and organochlorine compounds in the blubber of females and calves. These data indicate that calves quickly bioaccumulate contaminants by transplacental and lactational routes to concentrations that are in equilibrium with females. In comparisons between contaminant concentrations and patterns in the blubber of female blue and humpback whales, there were no significant differences in concentrations, but the proportions of some PCB congeners, HCH isomers, and DDT and its metabolites were different in the two baleen whale species. These may reflect differences in the diet of the two species, since fish comprise a large part of the diet of humpback whales and blue whales feed exclusively on euphausiid crustaceans (i.e. krill). PMID- 14749059 TI - The relative importance of water and diet for uptake and subcellular distribution of cadmium in the deposit-feeding polychaete, Capitella sp. I. AB - The impact of dietary and water exposure on the accumulation and distribution of cadmium (Cd) in subcellular components of the polychaete Capitella sp. I was investigated. Worms were exposed to either dissolved Cd alone ('Water-Only' treatments; WO) or diet-bound Cd alone ('Algae-bound Only' treatments; AO). Thus, WO worms were starved and AO worms were fed. Differential centrifugation was used to fractionate worm homogenates into debris- (DE), mitochondrial- (MI), microsomal- (MC) and cytosolic- (CY) fractions, and the concentration of Cd in these fractions was quantified by radiometric analysis using the specific activity of the (109)Cd radioisotope. Neither exposure route nor Cd exposure affected the total protein content in the worms. However, fed, but not starved worms relocated proteins from the mitochondria to the cytosol in response to Cd exposure. In systems without Cd addition, starvation likewise influenced the distribution of protein between mitochondria and cytosol. Cutaneous uptake and accumulation of Cd from the water was related to surface area while dietary uptake was influenced by the amount of sediment passing through the gut. Irrespective of exposure route, Cd was accumulated primarily in the debris fraction although comparisons between the treatments indicated that relatively less Cd was located in DE and more in MI, MC and CY fractions in worms in AO compared to worms in WO. Cd per unit protein in DE and MI fractions was twice as high as in MC and CY fractions in WO, whereas MC contained the highest Cd content followed by MI, DE and CY fractions in AO worms. Our results stress the importance of exposure route for the internal distribution and toxicity of Cd to deposit feeders such as Capitella sp. I. PMID- 14749060 TI - The effects of power station entrainment passage on three species of marine planktonic crustacean, Acartia tonsa (Copepoda), Crangon crangon (Decapoda) and Homarus gammarus (Decapoda). AB - Experiments have been undertaken exposing larval common shrimp (Crangon crangon) and lobster (Homarus gammarus) and adult copepods (Acartia tonsa) to the key stresses of entrainment within power-station cooling-water systems. The apparatus has enabled the testing of mechanical, thermal, chlorine and realistic pressure effects both alone and in combination, the range of stressors spanning the standard conditions found within a temperate coastal direct-cooled power station. Mechanical stresses affected only lobster larvae, pressure changes affected only the Acartia adults. Residual chlorine caused significant mortality of Acartia and shrimp larvae, but had no effect on lobster larvae even at 1 ppm. The temperature increment significantly affected all three species, with a synergistic effect on chlorine sensitivity in the shrimp larvae, but only temperatures higher than would be experienced in a normally-operating power station affected the copepods. The majority of individuals of each species would survive passage through a power station system under normal conditions. It is notable that, within the species tested, generalizations from the responses of one species to those of another are not valid. PMID- 14749061 TI - Implications of seasonal priming and reproductive activity on the interpretation of Comet assay data derived from the clam, Tapes semidecussatus Reeves 1864, exposed to contaminated sediments. AB - We explore the use of the clam Tapes semidecussatus Reeves 1864 as an indicator for the presence of potentially genotoxic substances in estuarine sediments. The limitations associated with the interpretation of Comet assay data (expressed as % DNA in tail) in terms of clam reproductive state, size (age) and thermal exposure history following laboratory acclimation are discussed. Hatchery-reared clams, subjected to ambient temperature fluctuations during growth, were exposed in vivo under laboratory conditions for three weeks to sediment samples collected from a polluted site and a "clean" reference site. The DNA damage observed in haemocytes, gill and digestive gland cells was significantly higher in animals exposed to contaminated sediment compared to those exposed to sediment from the reference site. The extent of DNA damage recorded was not correlated with size (age). Spawning was not observed during the experiment. Nevertheless, clams with well-developed gonads showed a statistically higher degree of DNA damage in gill and digestive gland cells- but not haemocytes, demonstrating an increased sensitivity to potential genotoxic compounds, possibly caused by impaired DNA repair capacity due to reproductive activity. Furthermore, the degree of DNA damage in clams exposed to contaminated sediments was higher in autumn and winter compared to spring and summer, suggesting an effect of seasonal priming. PMID- 14749062 TI - Crude oil bioremediation in sub-Antarctic intertidal sediments: chemistry and toxicity of oiled residues. AB - The effectiveness of fertilizers for crude oil bioremediation in sub-Antarctic intertidal sediments was tested over a one-year period in a series of ten (10) experimental enclosures. Chemical, microbial and toxicological parameters demonstrated the effectiveness of various fertilizers in a pristine environment where hydrocarbon degrading bacteria (HDB) had not been stimulated by previous accidental spills or human activities. The low temperature of seawater (3-4 degrees C) had no obvious effects on the HDB community and the bioremediation process. Over 90% of n-alkanes were degraded in the first six months and most light aromatics (2-3 rings) disappeared during the first year of observation. The toxicity of oiled residues (Microtox(R) SP) was significantly reduced in the first 6 months of the process, but it increased again in the last months of the experiment. One of the fertilizers containing fishbone compost enriched with urea, inorganic phosphorus and a lipidic surfactant reduced significantly the toxicity of oil residues in the last 3 months of the experiment. Interstitial waters collected below the oil slicks during the remediation showed no toxicity, and even stimulated Vibrio fischeri. When comparing all fertilizers to the control plots, a good correlation (r(2)=0.82) was found between the growth rate of HDB and the degradation rate of n-alkanes in the first 90 days of the experiment only indicating that fertilizers were efficient for at least 3 months but their beneficial effects were lost after 6 months. PMID- 14749063 TI - Preliminary evidence of the role of hydrogen peroxide in the degradation of benzo[a]pyrene by a non-white rot fungus Fusarium solani. AB - In order to study the enzymatic mechanisms involved in the successive steps of BaP degradation by a Deuteromycete fungus Fusarium solani, we developed an indirect approach by using inhibitors of enzymes. We used either specific inhibitors of peroxidases (i.e. salicylhydroxamic acid) and of cytochrome P-450 (i.e. piperonyl butoxyde) or inhibitors of both enzymes (i.e. potassium cyanide). Surprisingly, no expected decrease of BaP degradation was observed with most inhibitors tested. On the contrary, more BaP was degraded. Only butylated hydroxytoluene, which acts as a free radical scavenger, inhibited BaP degradation. The inhibition of these enzymes, which use H(2)O(2) as a cosubstrate, might have resulted in an increase of hydrogen peroxide availability in the fungal cultures. This enhancement could induce formation of reactive oxygen species (ROS) which might be the agents that initiate benzo[a]pyrene oxidation. This study proposed a hypothetic alternative metabolic pathway involved in PAH metabolism by Fusarium solani. PMID- 14749064 TI - Evaluation of factors influencing root-induced changes of copper fractionation in rhizosphere of a calcareous soil. AB - Major factors influencing the root-induced copper fractionation changes within the rhizosphere of maize, wheat, pea, and soybean seedlings were evaluated using a contaminated calcareous soil. The effects of acidification, alkalization, and introduction of root exudates were investigated by addition of acid, alkaline and root exudates from solution cultures, prior to incubation and copper fractionation. Raw and sterilized soils were compared for changes of copper fractionation in the rhizosphere using rhizoboxes with maize, wheat, pea and soybean seedlings. The results indicated that the general trend in considerable changes was similar among the plant species studied. The rhizosphere experienced a depletion of carbonate associated and organic bound copper along with an accumulation of exchangeable and Fe-Mn oxide bound copper. The resulting significant influence of root exudates on copper fractionation appears to have been produced through complexation rather than acidification or alkalization. The increase in exchangeable copper in rhizosphere was strengthened by microorganisms. PMID- 14749065 TI - A case study of contaminants on military ranges: Camp Edwards, Massachusetts, USA. AB - An extensive investigation at the Camp Edwards, Massachusetts Military Reservation (MMR) demonstrates that assessment of groundwater and soil contamination at military ranges can be limited primarily to explosive-related compounds such as RDX, HMX, perchlorate, TNT and their transformation products. A modified analytical method is recommended to expand the list of explosives and to improve the detection limits. Analyses of metals, VOCs, SVOCs, and TICs are unnecessary. Soil samples may require the analyses of PAHs and PCNs for burn areas. Camp Edwards, as one of the few military ranges that have been exhaustively investigated for contaminants, is an ideal point of departure for evaluating other ranges. The permeable site soils promote leaching of contaminants and inhibit biotic and abiotic transformations. Moreover, the site has experienced an unusual extent of activities in its more than ninety years of active use. The recommendations in this report are based on data obtained for more than 200 analytes from more than 15,000 environmental samples. PMID- 14749066 TI - Tissue metal levels in Muskrat (Ondatra zibethica) collected near the Sudbury (Ontario) ore-smelters; prospects for biomonitoring marsh pollution. AB - An examination of tissue metal levels in Sudbury-area muskrat (Ondatra zibethica) revealed that animals collected in the vicinity of the local ore-smelters contained elevated burdens of Cd and Ni in their liver and kidneys. Respective tissue concentrations averaged 2-fold and 3- to 6-fold higher than background values and are believed to reflect accumulations resulting from food chain contamination in regional marshes, including that reportedly characterizing Typha latifolia stands-their primary food source-and adherent sediments which may be consumed inadvertently while feeding. No evidence of site-influence or enhanced tissue metal levels was seen for Cu, Pb or Zn. While Cd : Ni accumulations were positively correlated in both the liver (r=0.78) and the kidneys (r=0.65), between-tissue comparisons indicated that hepatic : renal burdens were significantly correlated (r=0.75) only in the case of Ni. With the exception of 30-35% lower hepatic Zn levels in females relative to males within the Sudbury population, tissue metal levels did not vary according to sex or age class at either site. Our findings substantiate the potential of muskrat to serve as useful bioindicators/monitors of metal pollution in semi-aquatic environments. PMID- 14749067 TI - Constraints on the use of 137Cs as a time-marker to support CRS and SIT chronologies. AB - CRS and SIT are two (210)Pb-based models widely used in the radiometric dating of recent sediments. (210)Pb chronologies should be validated using at least one independent tracer, such as (137)Cs. This paper demonstrates that simple methods based on the identification of (137)Cs fallout peaks cannot provide a definitive support for CRS and SIT chronologies. Two main arguments will support this assertion: Firstly, the (137)Cs time-marks cannot support a CRS or SIT chronology if the derived sedimentation rates cannot explain the whole (137)Cs activity profile without postulating mixing. Secondly, the support by the (137)Cs time marks for a given CRS or SIT chronology cannot be considered as definitive if other dating models can equally explain the whole set of data, thereby producing a different chronology. Several case studies selected from the literature are used to support the present discussion. PMID- 14749068 TI - Assessing the cause of impacts on benthic organisms near Rouyn-Noranda, Quebec. AB - Sediments from lakes near Rouyn-Noranda, Quebec, contain elevated concentrations of several metals, including Cd, Cu, Pb and Zn. Amphipods, fingernail clams, mayflies and tanytarsid midges were absent, and sediment toxicity was observed in chronic tests with Hyalella in sediments from Lac Dufault, the lake closest to Rouyn-Noranda. Bioaccumulation by Hyalella demonstrated elevated bioavailability of Cd, Co, Cr, Pb and Tl, but only Cd was accumulated to levels close to the toxic threshold. Copper, which is regulated by Hyalella, was not elevated in these amphipods, but it was elevated in overlying water in the toxicity tests. Toxic effects in Lac Dufault sediments are probably caused primarily by Cd, at least in amphipods, with a possible minor contribution from Cu. An integrated assessment, including sediment chemistry, benthic community composition, sediment toxicity, metal bioaccumulation in benthos, and comparison of bioaccumulation and/or overlying water concentrations with threshold effect concentrations, provides the best indication of effects and their cause. PMID- 14749069 TI - Ostracoda and foraminifera as short-term tracers of environmental changes in very polluted areas: the Odiel Estuary (SW Spain). AB - The analysis of 17 cores collected in the Odiel Estuary (SW Spain) permits delimiting the recent evolution of this zone during the past decades and the influence of natural and anthropogenic factors on the distribution of Ostracoda and Foraminifera. In the upper estuary, the coincidence of acid waters, prolonged subaerial exposure, and coarse sediments may explain the absence or the disappearance of these microorganisms during the industrial period (1966-1985) in the major part of this area. In the lower estuary, sedimentary evolution and industrial wastes are the main factors influencing both the distribution and trends of the populations of these two groups. Finally, the main changes observed in the marine estuary are due to the sedimentary effects of the construction of two banks and the dredging of the main estuarine channel. PMID- 14749071 TI - Low molecular weight thiols in arsenic hyperaccumulator Pteris vittata upon exposure to arsenic and other trace elements. AB - Low molecular weight thiol-containing compounds have been reported to play an important role in metal detoxification and accumulation in some higher plants. The formation of these low molecular weight thiols in the recently discovered arsenic hyperaccumulator, Chinese Brake fern (Pteris vittata) upon exposure to arsenic and other trace metals was investigated. In addition to cysteine and glutathione, an unidentified thiol was observed in the plants exposed to arsenic, which was not found in the control. The concentration of the unidentified thiol showed a very strong and positive correlation with arsenic concentration in the leaflets. The unidentified thiol was low in rachises and undetectable in the roots for As-treated plants. Total and acid-soluble thiols were also measured and the results indicated that arsenic mainly stimulated the synthesis of acid soluble thiol in Chinese Brake. The investigations of other trace elements (Cd, Cu, Cr, Zn, Pb, Hg, and Se) showed that these elements were not accumulated in Chinese Brake to high levels and the synthesis of the unidentified thiol in the plant was not observed. Our study suggests that the unidentified thiol was induced specifically by arsenic and the distribution patterns of the unidentified thiol and arsenic in the plant were consistent, indicating that the synthesis of this compound was related to As exposure. PMID- 14749070 TI - Rainwater chemistry at the summit and southern flank of the Itatiaia massif, Southeastern Brazil. AB - Wet deposition and related rainwater chemistry were studied at the Itatiaia massif, on which is settled the Itatiaia National Park (INP). Samples were simultaneously collected on a weekly basis over 12 months, using automated wet and dry samplers, at the INP-Headquarters (INP-Hq; altitude=820 m) and the Itatiaia Plateau (It-Pt; altitude=2460 m). Conductivity, pH, Na(+), K(+), Mg(2+), Ca(2+), NH(4)(+), Cl(-), NO(3)(-) and SO(4)(2-) were determined in 36 rainwater samples. Volume-weighted mean (VWM) pH was lower at the INP-Hq (4.9) than at the It-Pt (5.3). Very strong correlation between Cl(-) and Na(+) was found for the INP-Hq (r=0.99). At the Itatiaia massif, SO(4)(2-), NO(3)(-), and NH(4)(+) comprised together about 60% of the total inorganic ions and appear to exert the major control on rainwater pH. PMID- 14749072 TI - Sulphur dioxide adsorption in Scots pine canopies exposed to high ammonia emissions near a Cu-Ni smelter in SW Finland. AB - Since 1994 the nickel-processing plant at the Cu-Ni smelter at Harjavalta, south west Finland, has emitted considerable amounts of NH(3) into the atmosphere. The effects of NH(3) emissions on nitrogen and sulphur deposition in throughfall and the foliar nutrient status were investigated in a Scots pine stand at 0.5 km distance. Bulk deposition, stand throughfall and percolation water (20 cm depth) samples were collected at 4-week intervals during 1992-1998. pH and the Ca, Mg, K, NH(4) and SO(4) concentrations were determined on the samples. NH(3) emissions have strongly increased the scavenging of SO(2) from the air in the pine stand, and the increased levels of N and S deposition were clearly evident as increased foliar N and S concentrations and larger needle size. The increased input of SO(4) into the forest floor was not associated with an increase in the leaching of Ca and Mg from the surface soil layers. PMID- 14749073 TI - Chemical coupling between ammonia, acid gases, and fine particles. AB - The concentrations of inorganic aerosol components in the fine particulate matter (PM(fine)< or =2.5 microm) consisted of primarily ammonium, sodium, sulfate, nitrate, and chloride are related to the transfer time scale between gas to particle phase, which is a function of the ambient temperature, relative humidity, and their gas phase constituent concentrations in the atmosphere. This study involved understanding the magnitude of major ammonia sources; and an up wind and down-wind (receptor) ammonia, acid gases, and fine particulate measurements; with a view to accretion gas-to-particle conversion (GTPS) process in an agricultural/rural environment. The observational based analysis of ammonia, acid gases, and fine particles by annular denuder system (ADS) coupled with a Gaussian dispersion model provided the mean pseudo-first-order k(S-1) between NH(3) and H(2)SO(4) aerosol approximately 5.00 (+/-3.77)x10(-3) s(-1). The rate constant was found to increase as ambient temperature, wind speed, and solar radiation increases, and decreases with increasing relative humidity. The observed [NH(3)][HNO(3)] products exceeded values predicted by theoretical equilibrium constants, due to a local excess of ammonia concentration. PMID- 14749074 TI - Cellular energy allocation in zebra mussels exposed along a pollution gradient: linking cellular effects to higher levels of biological organization. AB - Organisms exposed to suboptimal environments incur a cost of dealing with stress in terms of metabolic resources. The total amount of energy available for maintenance, growth and reproduction, based on the biochemical analysis of the energy budget, may provide a sensitive measure of stress in an organism. While the concept is clear, linking cellular or biochemical responses to the individual and population or community level remains difficult. The aim of this study was to validate, under field conditions, using cellular energy budgets [i.e. changes in glycogen-, lipid- and protein-content and mitochondrial electron transport system (ETS)] as an ecologically relevant measurement of stress by comparing these responses to physiological and organismal endpoints. Therefore, a 28-day in situ bioassay with zebra mussels (Dreissena polymorpha) was performed in an effluent dominated stream. Five locations were selected along the pollution gradient and compared with a nearby (reference) site. Cellular Energy Allocation (CEA) served as a biomarker of cellular energetics, while Scope for Growth (SFG) indicated effects on a physiological level and Tissue Condition Index and wet tissue weight/dry tissue weight ratio were used as endpoints of organismal effects. Results indicated that energy budgets at a cellular level of biological organization provided the fastest and most sensitive response and energy budgets are a relevant currency to extrapolate cellular effects to higher levels of biological organization within the exposed mussels. PMID- 14749075 TI - The study of metal contamination in urban soils of Hong Kong using a GIS-based approach. AB - The study of regional variations and the anthropogenic contamination by metals of soils is very important for environmental planning and monitoring in urban areas. An extensive survey was conducted in the highly urbanized Kowloon area (46.9 km(2)) of Hong Kong, using a systematic sampling strategy with a sampling density of 3-5 composite soil samples (0-15 cm) per km(2). Geochemical maps of 'total' metals (Cd, Cr, Cu, Ni, Pb and Zn) from strong acid extraction in the surface soils were produced based on geographical information system (GIS) technology. A significant spatial relationship was found for Ni, Cu, Pb and Zn in the soils using a GIS-based analysis, suggesting that these metal contaminants in the soils of the Kowloon area had common sources. Several hot-spot areas of metal contamination were identified from the composite metal geochemical map, mainly in the old industrial and residential areas. A further GIS analysis revealed that road junctions, major roads and industrial buildings were possible sources of heavy metals in the urban soils. The Pb isotope composition of the contaminated soils showed clear anthropogenic origins. PMID- 14749076 TI - The effect of heavy metal exposure on egg size, eggshell thickness and the number of spermatozoa in blue tit Parus caeruleus eggs. AB - In this study we compared the egg size, the eggshell thickness and the number of spermatozoa trapped on the perivitelline layer of the eggs in three blue tit (Parus caeruleus) populations across a heavy metal pollution gradient. No significant differences were observed in egg characteristics among study sites. Eggs from the two most polluted sites however, had significantly less spermatozoa on the perivitelline layer than eggs from the least polluted site. Our results suggest a significant effect of pollution on blue tit sperm concentrations in the infundibulum. Determining the number of spermatozoa on the egg membranes is relatively cheap and straightforward, which renders this technique a new and useful tool in biomonitoring. PMID- 14749077 TI - Spatial and temporal trends in surface water and sediment contamination in the Laurentian Great Lakes. AB - Data from recent sediment and surface water surveys have been collated and mapped to illustrate the spatial distribution of contaminants across the entire Great Lakes basin. Information from historical surveys, together with data from surface water monitoring programs in three major connecting channels, has also been collated in order to evaluate temporal trends. In general, Lakes Superior and Michigan exhibited the lowest levels of sediment contamination while Lake Ontario had the highest. Contaminants such as gamma-HCH (lindane) and dieldrin were ubiquitous in surface waters across the entire basin, which was indicative of atmospheric sources. The distribution of other compounds including hexachlorobenzene, octachlorostyrene and mirex indicated the presence of local sources within the watersheds of the connecting channels. Surficial sediment contamination was found to have decreased markedly since the late 1960s and 1970s. Similarly, surface water contamination decreased over the period 1986-1997 with concentrations of dieldrin, hexachlorobenzene, octachlorostyrene and mirex reduced by over 50%. However, the spatial distributions of both sediment and surface water contamination indicate that further effort is warranted in reducing local sources of contaminants, particularly in Lake Ontario. PMID- 14749078 TI - Sulphur, nitrogen and carbon content of Sphagnum capillifolium and Pseudevernia furfuracea exposed in bags in the Naples urban area. AB - The accumulation ability of the major elements sulphur, nitrogen and carbon by the moss Sphagnum capillifolium (Ehrh.) Hedw. and the lichen Pseudevernia furfuracea (L.) Zopf exposed in bags in Naples urban area,was investigated. Bags were exposed at the beginning of July 1999 and gathered in two subsequent moments: at the end of the dry season (after 10 weeks of exposure) and during the wet season (after 17 weeks of exposure), to include the effects of rainy conditions. Sulphur and N content of the lichen increased all over the exposure period, while the level of C did not change significantly either after 10 or 17 weeks of exposition. For the moss the S accumulation was limited to the dry period of exposure, whereas N and C content decreased with exposure. Results, in contrast with those obtained in a previous study on trace elements bioaccumulation [Adamo et al., Environmental Pollution, (2003) 122, 91-103], suggest that accumulation of gaseous pollutants is strongly influenced by biomonitor vitality and that lichen bags are a more reliable and effective tool for monitoring S, N and C atmospheric depositions in urban areas compared to moss bags, because of greater lichen resistance to dry and stressing conditions of urban environment. PMID- 14749079 TI - The need for ammonia abatement with respect to secondary PM reductions in Europe. AB - In Europe, secondary particulate matter (PM) comprises 50% or more of PM 2.5. To reduce PM concentrations requires lowering precursor emissions. Since the 1980s, SO(2) emissions have decreased by more than 60%, while particle concentrations have decreased less. NO(x) and NH(3) emissions have decreased slightly. The role of ammonia in particle formation is addressed here. It is shown that secondary PM concentrations can only be effectively reduced if ammonia emissions are decreased in much the same way as those of SO(2) and NO(x). PMID- 14749080 TI - Effect of sandblasting, grinding, polishing and glazing on the flexural strength of two pressable all-ceramic dental materials. AB - OBJECTIVES: During laboratory fabrication procedures and/or clinical adjustments, pressable materials: IPS Empress and Empress 2, may be ground, polished or sandblasted. These treatments may affect their strength by introducing microscopic flaws and defects. This study investigates the effect of these procedures on the mean flexural strength of these materials. METHODS: One hundred and forty disc specimens (14mmx1 mm) of IPS Empress and Empress 2 were prepared, and divided into seven groups of 20 specimens for each material. Groups were untreated, polished, polished and glazed, ground, ground and glazed, sandblasted, sandblasted and glazed. Surface roughness, mean biaxial flexural strength and Weibull modulus were appraised, and a scanning electron microscope was used to describe surface features. Statistical significance among groups of population was analysed using one-way Anova and Tukey's multiple comparison tests. RESULTS: Untreated and sandblasted groups showed significantly the highest roughness values, and polished the lowest for each material (p<0.05). Ground groups showed significantly lower roughness values than the sandblasted groups, and significantly higher roughness than the polished groups for each material (p<0.05). Polished groups for each material demonstrated significantly the highest mean flexural strength values (p<0.05). No significant difference in the mean strength values was found between untreated, sandblasted and ground groups for each material (p>0.05). Heat treatment had no effect on roughness or strength values of all treated groups of both materials. The Weibull modulus values for both materials varied with different treatments. They showed higher values for polished and untreated groups, and lower values for ground and sandblasted groups. CONCLUSIONS: Surface roughness may not be the only feature that determines strength. Other issues such as porosity, microstructural residual stresses, surface and bulk defects may also be pertinent. PMID- 14749081 TI - The role of organic tissue on the punch shear strength of human dentin. AB - OBJECTIVES: The tendency to fracture of human teeth has been attributed to numerous reasons, including pulpless tooth dehydration, tooth structure degeneration, and excessive spreader loading. To date, however, no consistent data has been gathered to ascertain the effect of organic tissue on the biomechanical properties of human dentin. This study explored the influences of organic tissue removal on the morphological changes and punch shear strength by means of low vacuum wet scanning electron microscope (Wet-SEM) and punch shear apparatus. METHODS: The crown dentin slices with a thickness of 0.45-0.50 mm were prepared from extracted human molars. These specimens were immersed in 5% NaOCl for 1, 3, and 6 h to remove the organic tissue. The dynamic morphological changes of these dentin slices after immersion were observed using Wet-SEM. In addition, the punch shear strength and stiffness of specimens were measured. RESULTS: The removal of organic tissue by 5% sodium hypochlorite treatment caused many cracks on the specimen surfaces. Intertubular and peritubular dentin were also affected, causing an abnormal configuration of dentinal tubule orifices. The 5% sodium hypochlorite treatment for 1, 3, and 6 h significantly reduced the punch shear strength of dentin. As the immersion time increased, the values of punch shear strength also diminished. CONCLUSIONS: The organic tissue loss of human dentin would affect the structure of intertubular and peritubular dentin and cause many cracks on the dentin surface. The punch shear strength of human dentin also reduced after removal of organic tissue. PMID- 14749082 TI - Determinants of in vitro gap formation of resin composites. AB - OBJECTIVES: To investigate whether polymerization shrinkage, flow, modulus, and bond strength influence marginal gap formation of resin composite restorations in vitro. METHODS: Eleven proprietary resin composites were studied. Shrinkage was measured by the 'bonded-disk method' (n=3). Flow was measured as the diameter of a constant volume of resin composite to which a load of 20 N had been applied for 60 s (n=3). Modulus was measured in 3-point bending (n=6). Bond strength mediated to human dentin by an adhesive system was measured in shear (n=6). Gap formation was measured in a light microscope along the margins of all-dentin, butt-joint cavities restored with the adhesive system and each resin composite and expressed as the widest gap in percent of the cavity diameter (n=6). RESULTS: Significant differences were found between the resin composites regarding all determined properties. A significant linear correlation was found between flow and gap formation (r=-0.68, P<0.025). Three-dimensional regression analysis showed a significant correlation between polymerization shrinkage (X(1)), flow (X(2)), and gap formation (r=0.79, P(1)<0.05, P(2)<0.005). The highest coefficient of correlation was found when the first part of the polymerization shrinkage (from 0 to 10 s) was disregarded (r=0.90, P(1)<0.005, P(2)<0.0005). CONCLUSIONS: With the adhesive system used, polymerization shrinkage and flow were found to be significant determinants of gap formation around resin composite restorations in vitro. PMID- 14749083 TI - Determination of fluoride ions released from resin-based dental materials using ion-selective electrode and ion chromatograph. AB - OBJECTIVES: The aim of this study was to investigate the quantities of total and free fluoride ions released from four resin-based composites containing fluoride (Heliomolar (HE), UniFil S (UF), Beautifil (BE) and Reactmer paste (RP)) and one glass ionomer cement (Ketac-Fil Plus Aplicap (KF)). METHODS: Five disk specimens of each material were prepared and the fluoride release was measured at 1, 2, 4, 7, 14, 21 and 28 days. The amounts of total and free fluoride ions released from materials were measured by using an ion-selective electrode (ISE) and ion chromatography (IC). Data were statistically analyzed using a t-test at a significant level of 0.05. RESULTS: For HE, fluoride release is undetectable using the ISE but was detected by the IC method. There were no significant differences in the free and total fluoride ions released from UF and BE (p>0.05). For RP, the free fluoride ions were significantly lower than the total fluoride ions after 4 days (p<0.05). KF showed significant difference between total and free fluoride ions at 7 and 28 days (p<0.05). CONCLUSIONS: The nature of the fluoride incorporated into dental materials affected the amounts of free and total fluoride ions released from materials. PMID- 14749084 TI - Role of mechanical environment and implant design on bone tissue differentiation: current knowledge and future contexts. AB - OBJECTIVES: To evaluate published evidence related to bone reactions to varying loading regimens and the impact of implant design on bone tissue differentiation. DATA AND SOURCES: The literature was searched for original research articles relating effect of mechanical environment on bone tissue and effects of implant design on bone biomechanics and marginal bone reactions using MEDLINE and manual tracing of references cited in key papers otherwise not elicited. STUDY SELECTION: Current literature on biomechanics of bone and dental implants as main focus and pertinent to key aspects of the review. CONCLUSIONS: Implant design influences force transmission characteristics in peri-implant bone, but not the time-dependent marginal bone reactions. Mechanical signals affect bone tissue differentiation. Therefore, it is essential to control biomechanical loads on implants to maintain osseointegration and/or to promote early bone-implant interface healing. PMID- 14749085 TI - Mathematical coupling can undermine the statistical assessment of clinical research: illustration from the treatment of guided tissue regeneration. AB - OBJECTIVES: Previous periodontal literature has shown that there is a strong relationship between treatment effects, such as guided tissue regeneration (GTR), and baseline disease severity. However, relating change to baseline values using correlation or regression is methodologically flawed due to mathematical coupling, where the statistical procedure of testing the null hypothesis-that the coefficient of correlation or slope of regression is equal to zero-becomes erroneous. The aim of this study is to investigate if baseline disease severity is genuinely associated with the treatment outcome of intrabony defects using GTR after adjustment for mathematical coupling. In particular, we seek to demonstrate the potential effect that mathematical coupling has in distorting the results from the statistical analyses of trials of dental treatment, using data from the periodontal literature on GTR. The erroneous results arising from the use of simple correlation and regression techniques to analyse this association will be demonstrated, also the methodological flaw where the statistical procedure tests the null hypothesis-that the coefficient of correlation or the slope of regression is equal to zero. METHODS: Three main periodontal journals were electronically and manually searched to extract the data for the clinical outcomes of pocket probing depth (PPD) and lifetime cumulative attachment loss (LCAL) in the studies using GTR. The relationship between clinical outcomes and baseline measurements were reanalysed using Oldham's method and the variance ratio test. RESULTS: The results of these analyses were compared with those from the papers where the authors used the standard approach of correlation or regression. This shows that mathematical coupling caused spurious correlations between baseline disease severity and treatment effect. Ten out of 12 studies for PPD and nine out of 14 for LCAL initially claimed a significant positive relationship; after using either of the more appropriate statistical methods of adjustment, only three correlations in each group of studies remained significant. CONCLUSIONS: Previous evidence suggesting an association between baseline disease severity and treatment effect for GTR is challenged and therefore needs to be critically reviewed. All future clinical research should avoid using mathematically coupled data in correlation or regression analysis. In seeking to examine the bivariate association between baseline and subsequent change, Oldham's method is recommended. PMID- 14749086 TI - Ratio variables in regression analysis can give rise to spurious results: illustration from two studies in periodontology. AB - OBJECTIVES: For over a century, statisticians have highlighted concerns about the inappropriate use of ratio variables in correlation and regression analysis. However, little attention has been paid to these concerns in medical and dental research. The use of ratio variables in correlation and regression analysis can give rise to spurious results due to inappropriate model specification and mathematical coupling, leading to serious misinterpretation of data and consequently to incorrect study conclusions. METHODS: Data were reanalysed from two recently published articles: one on the efficacy of guided tissue regeneration on root coverage; the other a randomised controlled trial comparing three surgical approaches in the treatment of periodontal infrabony defects. The reanalysis was performed to examine whether the assumptions behind the correlation/regression analyses have been seriously violated in these two studies, and to see if the interpretation of results is tenable. RESULTS: Use of ratio variables seriously violated the assumptions underpinning the statistical methods utilised in these two studies, and consequently the conclusions were substantially misleading. Recommendations made in these studies were not tenable. CONCLUSIONS: The reanalyses illustrate how the inappropriate use of ratio variables remains prevalent in dental research, leading to incorrect interpretation of the evidence. This emphasises the need for collaboration between clinicians and statisticians to avoid the risk of yielding erroneous conclusions from flawed statistical analyses. PMID- 14749087 TI - Effect of different crown contours on periodontal health in dogs. Microbiological results. AB - OBJECTIVES: In some clinical circumstances, i.e. in cases when the upper anterior region has to be restored by prosthetic means, it is necessary to place the margins of crowns and fixed partial dentures subgingivally. In addition, in periodontally compromised patients the restoration sometimes has to be overcontoured in order to replace the lost interdental papilla. The overcontoured crown margin may influence the subgingival bacterial composition. Therefore, the aim of the present investigation was to evaluate the effect of three different subgingival crown contours in dogs on the composition of the subgingival microbiota. METHODS: In four adult beagle dogs the second and third premolars were prepared in three quadrants and restored with single gold crowns. The unprepared second and third premolars in the last quadrant served as controls. The crowns had three different emergence profiles including a normal contour, a 30 degrees and a 50 degrees over-contour. During the entire study period, professional oral hygiene was performed seven times a week. Microbiological samples were harvested from four sites of test and control teeth (mesial, distal, buccal and lingual) at baseline, after 3 months, and after 5 months. RESULTS: The microbiological analysis (DNA-DNA hybridization technique) of the subgingival microbial flora revealed a dominance of P. intermedia, T. denticola and C. showae in all test and control groups at baseline. At three months, the total amount of bacteria increased and a broader variety of bacterial species could be detected. The detection frequency of most bacterial species increased from baseline to the 5-month evaluation. The frequency of detection of some species was higher in the 30 degrees and 50 degrees overcontoured test groups compared to the normal contour group and to the natural teeth. CONCLUSIONS: It can be concluded within the limits of this investigation that overcontoured gold crowns placed subgingivally have only slight effects on the microbiological composition in dogs when an intensive oral hygiene regimen was executed. PMID- 14749088 TI - Effects of CO2 laser on fluoride uptake in enamel. AB - OBJECTIVES: The objective of this study was to investigate the effects of CO(2) laser on fluoride uptake in the loosely- and firmly-bound forms in enamel. METHODS: Five human molars were cut into halves before being treated with 2.0% NaF topical gel. Each half had three windows on the enamel surface, including one control and two experimental windows irradiated by two laser therapies. One half of each tooth was treated with 1 M KOH solution to remove the loosely-bound fluoride (calcium fluoride). A tooth section was obtained from each window and the relative fluorine concentration was analyzed with Secondary Ion Mass Spectrometry (SIMS). The morphology of the enamel surfaces in the windows was examined using an Environmental Scanning Electron Microscope (ESEM). RESULTS: Significant laser-induced increases in the uptake of fluoride were revealed in both loosely-bound and firmly-bound apatitic fluoride, with both laser treatments (all p<0.001). Calcium fluoride-like deposits on the enamel surfaces receiving the combined laser-fluoride treatment were revealed by ESEM. CONCLUSIONS: The finding of this study substantiated the laser effect in increasing the fluoride uptake into enamel. PMID- 14749089 TI - Letter to the editor "Efficacy of anti-retraction devices in preventing bacterial contamination of dental unit water lines" [Berlutti F, Testarelli L, Vaia F, De Luca M, Dolci G, Journal of Dentistry 31 (2003) 105-110]. PMID- 14749091 TI - Do antidepressants regulate how cortisol affects the brain? AB - Although the effects of antidepressants on glucocorticoid hormones and their receptors are relevant for the therapeutic action of these drugs, the molecular mechanisms underlying these effects are unclear. Studies in depressed patients, animals and cellular models have demonstrated that antidepressants increase glucocorticoid receptor (GR) and mineralocorticoid receptor (MR) expression and function; this, in turn, is associated with enhanced negative feedback by endogenous glucocorticoids, and thus with reduced resting and stimulated hypothalamic-pituitary-adrenal (HPA) axis activity. In a series of studies conducted over the last few years, we have shown that antidepressants modulate GR function in vitro by inhibiting membrane steroid transporters that regulate the intracellular concentration of glucocorticoids. In this paper, we will review the effects of membrane steroid transporters and antidepressants on corticosteroid receptors. We will then present our unpublished data on GR live microscopy in vitro, showing that ligand-induced translocation of the GR starts within 30 seconds and is completed within minutes. Furthermore, we will present our new data using an in situ brain perfusion model in anaesthetised guinea-pigs, showing that entry of cortisol to the brain of these animals is limited at the blood brain barrier (BBB). Finally, we will present a comprehensive discussion of our published findings on the effects of chemically unrelated antidepressants on membrane steroid transporters, in mouse fibroblasts and rat cortical neurones. We propose that antidepressants in humans could inhibit steroid transporters localised on the BBB and in neurones, like the multidrug resistance p glycoprotein, and thus increase the access of cortisol to the brain and the glucocorticoid-mediated negative feedback on the HPA axis. Enhanced cortisol action in the brain might prove to be a successful approach to maximise therapeutic antidepressant effects. PMID- 14749092 TI - Mindfulness-based stress reduction in relation to quality of life, mood, symptoms of stress and levels of cortisol, dehydroepiandrosterone sulfate (DHEAS) and melatonin in breast and prostate cancer outpatients. AB - OBJECTIVES: This study investigated the relationships between a mindfulness-based stress reduction meditation program for early stage breast and prostate cancer patients and quality of life, mood states, stress symptoms, and levels of cortisol, dehydroepiandrosterone-sulfate (DHEAS) and melatonin. METHODS: Fifty nine patients with breast cancer and 10 with prostate cancer enrolled in an eight week Mindfulness-Based Stress Reduction (MBSR) program that incorporated relaxation, meditation, gentle yoga, and daily home practice. Demographic and health behavior variables, quality of life, mood, stress, and the hormone measures of salivary cortisol (assessed three times/day), plasma DHEAS, and salivary melatonin were assessed pre- and post-intervention. RESULTS: Fifty-eight and 42 patients were assessed pre- and post-intervention, respectively. Significant improvements were seen in overall quality of life, symptoms of stress, and sleep quality, but these improvements were not significantly correlated with the degree of program attendance or minutes of home practice. No significant improvements were seen in mood disturbance. Improvements in quality of life were associated with decreases in afternoon cortisol levels, but not with morning or evening levels. Changes in stress symptoms or mood were not related to changes in hormone levels. Approximately 40% of the sample demonstrated abnormal cortisol secretion patterns both pre- and post-intervention, but within that group patterns shifted from "inverted-V-shaped" patterns towards more "V-shaped" patterns of secretion. No overall changes in DHEAS or melatonin were found, but nonsignificant shifts in DHEAS patterns were consistent with healthier profiles for both men and women. CONCLUSIONS: MBSR program enrollment was associated with enhanced quality of life and decreased stress symptoms in breast and prostate cancer patients, and resulted in possibly beneficial changes in hypothalamic pituitary-adrenal (HPA) axis functioning. These pilot data represent a preliminary investigation of the relationships between MBSR program participation and hormone levels, highlighting the need for better-controlled studies in this area. PMID- 14749093 TI - Exposure to bisphenol A during gestation and lactation causes loss of sex difference in corticotropin-releasing hormone-immunoreactive neurons in the bed nucleus of the stria terminalis of rats. AB - It has been suspected that endocrine disrupters induce abnormal differentiation and development of reproductive organs. In the present study, we examined whether exposure to bisphenol A (BPA), a known endocrine disrupter, during gestation and lactation affects sex difference in the number of corticotropin-releasing hormone immunoreactive neurons (CRH neurons) in the preoptic area (POA) and the bed nucleus of the stria terminalis (BST). For that purpose, pregnant female Wistar rats (n=8-11 per treatment group) were treated with either 0.1% ethanol (control group) or 10 mg/l BPA (BPA group) dissolved in their drinking water until their offspring were weaned. In the control group, we confirmed a previous report that the POA of female rats contained significantly more CRH neurons than that of male rats (p<0.05). This significant sex difference was also evident in the BPA group, indicating that BPA exposure used in the present study had no effect on the sex difference in CRH neurons in the POA. We also found in the control group that the BST of female rats contained significantly more CRH neurons (p<0.05) than that of male rats. However, this significant sex difference was not observed in the BPA group (p>0.05), suggesting that BPA exposure affected the sex difference in CRH neurons in the BST. Since there was no statistically significant difference in the number of CRH neurons between the control and the BPA group, irrespective of the sex, the results suggested that a loss of sex difference in CRH neurons was due to both an increase in CRH neurons in male rats and a decrease in CRH neurons in female rats. The present study indicates that there is a significant sex difference in the number of CRH neurons in the BST as well as in the POA and that exposure to BPA during gestation and lactation causes a loss of this sex difference in the rat BST, but not in the POA. We suggest that CRH neurons in the BST are more susceptible to endocrine disrupters than those in the POA, irrespective of the sex. PMID- 14749094 TI - Chronic pregnenolone effects in normal humans: attenuation of benzodiazepine induced sedation. AB - Pregnenolone is the major steroid precursor in humans. It is also a "neurosteroid" and possesses intrinsic behavioral and brain effects in animals, affecting the GABA(A) and other receptors. In two preliminary studies, we sought to characterize its tolerability and psychotropic effects in humans. In Study 1, 17 normal volunteers received pregnenolone and placebo for 4 weeks each (15 mg PO per day x2 weeks followed by 30 mg PO per day x2 weeks, vs. placebo x4 weeks) in a within-subject, double-blind, cross-over design, with a 4 week drug-free washout period separating the two arms. Subjects' behavioral responses were assessed at the beginning and end of the 4-week pregnenolone arm and the 4-week placebo arm. Pregnenolone was generally well-tolerated but, by itself, had no significant effects on mood, memory, self-rated sleep quality or subjective well being. In Study 2, 11 subjects from Study 1 received a single dose of diazepam (0.2 mg/kg PO) immediately following completion of Study 1 in order to assess, in a between groups design, the impact of 4-weeks' pre-treatment with pregnenolone (N=5) vs. placebo (N=6) on the acute sedative, amnestic and anxiolytic effects of this benzodiazepine. Pregnenolone-pretreated subjects showed significantly less sedation following diazepam (p<0.03); this effect was clinically apparent. Diazepam's amnestic effects were non-significantly attenuated, and ratings of anxiety were unaffected. These pilot data, based on small samples, raise the possibility that chronically administered pregnenolone antagonizes certain acute effects of benzodiazepines and may enhance arousal via antagonist or inverse agonist actions at the benzodiazepine/GABA(A) receptor complex. Further larger scale studies, utilizing a broader range of doses and experimental conditions, are warranted. PMID- 14749095 TI - Altered hypothalamic-pituitary-adrenocortical function in rhesus monkeys (Macaca mulatta) with self-injurious behavior. AB - Individually housed rhesus monkeys sometimes spontaneously develop self-injurious behavior (SIB) in the form of self-directed biting that, on occasion, results in severe tissue damage and mutilation. We previously demonstrated lower levels of plasma cortisol in rhesus monkeys with a history of self-wounding (SW) when compared to non-wounders (NW). Furthermore, cortisol levels were negatively correlated with rates of self-directed biting. The present study was designed to further characterize the relationships between hypothalamic-pituitary adrenocortical (HPA) activity, self-wounding, and self-directed biting. Basal 24 h urinary free cortisol excretion, the urinary free cortisol response to a low dose of dexamethasone, and the plasma cortisol response to ACTH were examined in 24 individually housed rhesus monkeys, based on wounding history, i.e. the presence/absence of a veterinary record of self-wounding, and current rates of self-directed biting, i.e. the median split of self-directed biting frequency (independent of wounding status). There were no reliable group differences on any of the physiological measures when analyzed by wounding history. However, the plasma cortisol response 30 min post-ACTH stimulation was significantly correlated with wounding recency, such that lower responsivity was associated with more recent wounding episodes. When the results were analyzed on the basis of biting frequency, high frequency biters (HFB) compared to low frequency biters (LFB) showed decreased HPA negative feedback sensitivity to dexamethasone and a trend towards an attenuated plasma cortisol response to ACTH stimulation. These findings suggest that SIB in socially reared monkeys is associated with complex changes in HPA axis function that are related to the expression of the pathology, i.e. self-directed biting, and to the recency of a wounding episode. It remains to be determined whether humans who exhibit SIB show similar alterations in HPA function. PMID- 14749096 TI - Differences in cortisol awakening response on work days and weekends in women and men from the Whitehall II cohort. AB - It is thought that the salivary cortisol awakening response can serve as a reliable marker of hypothalamic-pituitary-adrenocortical activity. Response magnitude might be influenced by stress, but results of studies on work-related stress have been inconclusive. Non-compliance with the sampling schedule is a potential confounding factor that has rarely been controlled. The objective of the study was to determine whether the cortisol awakening response is greater on a work day than on a weekend day, and whether responses vary with gender and socioeconomic position. Compliance was controlled by excluding participants who reported a delay of more than 10 min between waking up and taking the first saliva sample. Data were collected from 196 men and women aged 47 to 59 years drawn from the Whitehall II cohort of British civil servants, with free salivary cortisol sampled immediately after waking up and 30 min later on 2 days. Data on stress, perceived control and happiness over the remainder of the day, and on sleep quality, time of waking, and health behaviour were also obtained. The awakening response was lower in non-compliant participants than in compliant ones, so non-compliant participants were excluded from further analyses. Salivary cortisol levels on waking did not differ by gender or socioeconomic position, or between work and weekend days. However, the cortisol awakening response (defined as the difference between waking and 30 min later) was greater on work than weekend days (mean increases 10.5 and 3.7 nmol/l, P < 0.001). On the work day, women showed larger increases than men (P = 0.011), but there were no gender differences on the weekend day. Across both days, lower socioeconomic position was associated with a larger cortisol awakening response (P = 0.014). Time of waking up was not related to the cortisol awakening response on either day. Participants rated themselves as more stressed, less in control, and less happy over the remainder of the work than weekend day. These results indicate that the cortisol awakening response occurs on both work and non-work days, but that anticipation of the working day is associated with an enhanced response. Cortisol output over the early part of the day may be particularly sensitive to the influence of chronic stress and its anticipation, especially in women. PMID- 14749097 TI - Exposure to retrieval cues improves retention performance and induces changes in ACTH and corticosterone release. AB - Memory retrieval can be facilitated by pretest exposure to cues associated with the original training. The present series of experiments was aimed at investigating whether the effectiveness of the retrieval cues might be due to their emotional value and thus be associated to a particular pattern of activation of stress systems. Therefore, the effects of exposing rats to different cueing conditions were investigated both on retention performance and on the level of different stress hormones (ACTH, corticosterone and glucose; the latter as an indirect index of adrenergic/sympathetic nervous system activation). Rats trained in a brightness avoidance discrimination task exhibited an enhancement of the retention performance following exposure to the light discriminative stimulus when delivered 1-day after training and not after 21 days, while exposure to contextual cues led to opposite effects on the retention performance, confirming our previous results. Analyses of the level of stress hormones at the time of testing indicated that when the retrieval cues were effective at the behavioral level, cued rats exhibited higher ACTH plasmatic levels than controls, but did not differ in their glucose or corticosterone levels. Further experiments showed that one day after training, both ACTH and corticosterone levels were elevated in light-cued rats if hormone samples were taken 15 min after cueing. These results show that exposure to an effective retrieval cue is accompanied by the activation of the hypothalamus-pituitary adrenal axis. The possible involvement of the Corticotropin Releasing Factor at the level of the hypothalamus and amygdala (particularly the central nucleus) on the facilitating effect on retention performance following exposure to aversive training-associated cues is discussed. The present results strengthen the notion that emotion can interact with retrieval processes. PMID- 14749098 TI - Correlation between serum levels of interleukin-4 and alexithymia scores in healthy female subjects: preliminary findings. AB - Some studies suggest that inaccuracy in recognizing and describing emotional states, combined with a highly descriptive mode of expression, as in alexithymia, may influence the immune response. We therefore investigated in healthy women the relationship between alexithymia and circulating levels of IL-1, IL-2 and IL-4. Seventeen mentally and physically healthy women aged between 20 and 25 years completed psychological questionnaires to assess alexithymia (Toronto Alexithymia Scale: TAS) and depressed mood (Hospital Anxiety and Depression Scale: HAD). Serum concentrations of IL-1, IL-2 and IL-4 were measured by ELISA. We found a significant positive correlation between serum levels of IL-4 and TAS score (r=0.55; p=0.021) and between factor 1 of the TAS (difficulty in identifying feelings) and IL-4 (r=0.57; p=0.017) while serum IL-1 and IL-2 were not detected in ten and six patients, respectively. Although there was a significant correlation between age and IL-4 levels, a linear regression with BMI, age, depressed mood and TAS as independent variables showed that only alexithymia could predict significantly increased levels of IL-4. Alexithymia and difficulty in identifying feelings could be associated with increased levels of IL-4 which may result in chronic impairment of pro/anti-inflammatory cytokine balance with psychological and somatic consequences. Nevertheless, these intriguing findings would deserve replication and extension in a larger sample of subjects. PMID- 14749099 TI - Letter to the Editor: Two formulas for computation of the area under the curve represent measures of total hormone concentration versus time-dependent change. A Letter to the Editor in response to J.C. Preussner et al. (2003) Psychoneuroendocrinology 28, 916-931. PMID- 14749101 TI - Determination of aniline and related mono-aromatic amines in indoor air in selected Canadian residences by a modified thermal desorption GC/MS method. AB - A modified thermal desorption method to determine low levels of aniline and other related mono-aromatic amines (MAAs) in residential air is described. The method was successfully applied to the determination of levels of these chemicals in residential air from 69 selected homes in two Canadian regions. Instrument detection limits of between 0.02 and 0.06 ng per thermal desorption tube were achieved for aniline and other MAAs. In the absence of environmental tobacco smoke (ETS) the mean concentrations of aniline in both indoor air and outdoor air were similar, at about 0.01 microgram/m3 after correction with field blanks. There was however a statistically significant difference in aniline concentrations between field blanks and indoor or outdoor air samples. No significant difference of aniline levels in the two study regions was observed. A clear link between aniline concentrations in indoor air and smoking activities inside homes was evident. Analysis of cigarette leaves indicated that aniline was most likely formed due to combustion during smoking. Shoe polishing was identified as another source of aniline in indoor air. PMID- 14749102 TI - Inhibition of seawater on bisphenol A (BPA) degradation by Fenton reagents. AB - To investigate bisphenol-A (BPA) degradation in seawater using Fenton reagents, changes in the BPA recovery and in the concentration of BPA metabolite, BPA-o quinone in the three water samples; BPA free deionizad water (control water), 3% aq. NaCl and seawater as a function of time after BPA fortification in the presence of radical oxygen species (ROS) at 20 degrees C were investigated. The BPA recoveries were lower in both 3% aq. NaCl and seawater than in the control water. The BPA recovery in aq. NaOCl decreased as a function of NaOCl concentration, indicating that BPA could be degraded by the potent radical ion (OCl-) at the concentration of above 2 microM. A BPA metabolite, BPA-o-quinone was formed in all the water samples after addition of ROS which was produced by Fenton reaction (reaction of 0.11 M H2O2 and 0.44 mM FeCl3.6H2O). These results indicated that BPA degradation could occur by an addition of ROS and further accelerated by the formation of OCl- in salt containing water samples. BPA recovery was the highest in seawater immediately after addition of Fenton reagents and the amount of BPA-o-quinone was very low, which suggests that seawater possesses an inhibitory system on BPA degradation. There was a positive correlation (p<0.01) between the fortified iron concentration and turbidity in seawater. Turbidity might be originated from iron-binding substances. Degradation threshold of BPA was observed when Fenton reaction was employed in seawater fortified with high amount of BPA. The present study suggested that iron trapping caused an inhibition on BPA degradation by Fenton reagents. PMID- 14749103 TI - Preliminary evaluation of PAH sorptive changes in soil by Soxhlet extraction. AB - This study was conducted to evaluate the influence of sorbent modification by synthetic, chemical/thermal weathering on the sorptive behavior of polycyclic aromatic hydrocarbons (PAHs). A clean sandy-clay-loam soil was subjected to Soxhlet extraction and PAH sorptive phenomena were evaluated based on quantity and quality changes in soil organic matter (SOM) and clay minerals. Critical changes in sorption capacity were found to depend on the initial PAH concentrations. Above 7 mg/l, weathering increased the PAH in comparison to that of unmodified soil, whereas it decreased when applied below this concentration. Similarly, less PAH was desorbed from the altered soil when PAH was applied above 7 mg/l. Therefore, when PAH was applied below 7 mg/l, quantitative reduction of sorbent amount (i.e., SOM and clay minerals) by soil weathering governed PAH sorptive behavior. However, when the PAH was applied above the critical limit, qualitative modifications in the sorbents facilitated an opposite trend. Sorbent swelling, removal of competing compounds, and possible changes in surface characteristics by Soxhlet extraction, together with increased concentration gradient effects were factors that resulted in dissimilar PAH sorptive phenomena, pivoting at the critical concentration. PMID- 14749104 TI - Metal contamination of farming soils affected by industry. AB - The contents of nine elements (As, Cd, Cr, Cu, Hg, Ni, Pb, Sb and Zn) have been assayed in the farming soils of Suszec commune (southern Poland). This area is affected by the main industrial centre of Poland (the Upper Silesian Industrial Region), the Czech Republic (Trzyniec smelter) and local contamination sources (coal mine). The contamination of the soils was assessed on the basis of geoaccumulation index, enrichment factor, contamination factor and degree of contamination. The tests revealed elevated contents of cadmium, lead, arsenic, antimony and mercury. The contents of the other elements were similar to the levels in the Earth's crust or pointed to metal depletion in the soil (EF<1). PMID- 14749105 TI - Effects of Se and Zn supplementation on the antagonism against Pb and Cd in vegetables. AB - The antagonistic effects of supplementation of Zn and Se to the soil on vegetables were studied in this work. In the pot experiment, Se (Se4+) and Zn (Zn2+) were applied, respectively, to the soil, in which the Chinese cabbage (Brassica rapa) and the lettuce (Lactuca sativa L.) were planted. As a result, Se and Zn were enriched evidently in the two vegetables. The contents of Pb and Cd in the two vegetables were decreased markedly while contents of some healthy mineral elements, like Mn and Mg, were increased to some extent when Se and Zn were applied. The antagonism of Se and Zn against Pb and Cd in plants was suggested. The farmland experiment on the lettuce was conducted to explore further the effect of supplementation of Zn and Se under the actual field conditions. Result came out to be that the enrichment of Zn and Se restrained the accumulation of Pb and Cd in the lettuce remarkably, as well as enhanced the absorption of some other nutritional elements, like Fe, Mn, Cu, Ca and Mg. Therefore, application of Se and Zn was proved to be an effective and feasible method to improve trace elements nutrition in the vegetables. PMID- 14749106 TI - Heavy metals monitoring at a Mediterranean natural ecosystem of Central Italy. Trends in different environmental matrixes. AB - The study deals with the evaluation of the impact of heavy metal pollution on a Mediterranean natural ecosystem, and presents the results derived from a monitoring of heavy metals in different environmental matrixes (atmospheric dry depositions, suspended particulate matter (SPM) and stemflow of forest trees). Two sites in Castelporziano Presidential Estate (Rome), one internal and one near the sea-side, were chosen in order to assess the differences in pollutant load. Results showed that heavy metal contamination can arise from local anthropogenic activities, in particular road traffic, and long-range pollution, from industrial and artisan activities near Rome. PMID- 14749107 TI - Heavy metals in mullet, Liza abu, and catfish, Silurus triostegus, from the Ataturk Dam Lake (Euphrates), Turkey. AB - The distribution of some heavy metals in three different organs of mullet, Liza abu, and catfish, Silurus triostegus, from Ataturk Dam Lake located on Euphrates (Turkey) was studied. Co and Mo concentrations were below limits of detection in all fish organs, whereas Ni was also below limits in organs of mullet. The metal accumulation in the liver and gill of L. abu and S. triostegus was found to be quite high in comparison with the muscle. In general, the concentrations are similar to those previously observed on other fish studied in Ataturk Dam Lake and lower than those determined in Tigris River. The analysed metals (Co, Cu, Fe, Mn, Mo, Ni and Zn) were found in fish muscle at mean concentrations under the permissible limits proposed by FAO. PMID- 14749108 TI - Wintertime indoor air levels of PM10, PM2.5 and PM1 at public places and their contributions to TSP. AB - From 26 October 2002 to 8 March 2003, particulate matter (PM) concentrations (total suspended particles [TSP], PM10, PM2.5 and PM1) were measured at 49 public places representing different environments in the urban area of Beijing. The objectives of this study were (1) to characterize the indoor PM concentrations in public places, (2) to evaluate the potential indoor sources and (3) to investigate the contribution of PM10 to TSP and the contributions of PM2.5 and PM1 to PM10. Additionally, The indoor and outdoor particle concentrations in the same type of indoor environment were employed to investigate the I/O level, and comparison was made between I/O levels in different types of indoor environment. Construction activities and traffic condition were the major outdoor sources to influence the indoor particle levels. The contribution of PM10 to TSP was even up to 68.8%, while the contributions of PM2.5 and PM1 to PM10 were not as much as that of PM10 to TSP. PMID- 14749109 TI - Geomorphological evolution and environmental reclamation of Fusaro Lagoon (Campania Province, southern Italy). AB - Analysis of morphological, geological and environmental characteristics of the Fusaro Lagoon has shown the present degraded condition of the lagoon and the perilagoon area. The lagoon developed during the mid-Holocene within a wide marine bay confined between the coastal volcanic structures of Mt. Cuma to the north and Torregaveta to the south in the western part of the Phlegrean Fields. Subsequently, the bay was gradually filled with pyroclastic materials from phlegrean eruptive vents and sediments carried by the rivers Volturno and Clanis, thus, creating an open lagoon. It then evolved into a partially closed lagoon due to the formation of a continuous littoral spit during the late Holocene, probably wider than the present-day one and surrounded by marshlands. Finally, the total closure of the lagoon took place in the Graeco-Roman period, following the stabilization of the dune ridge, and it assumed a shape similar to present-day one only towards the end of the 18th century. Between the Roman period and 1941, three lagoon channels were opened in order to avoid the frequent environmental crises which continue to affect, although for different reasons, this salt-water basin. The basin has been exploited for more than 2000 years not only for mollusc culture and pisciculture, but also for the maceration of hemp and flax. In the 1980s, in order to reduce the effects of the environmental crises, dredging of the lagoon bottom has been carried out, altering the hydrogeological equilibrium and that of the ecosystem. Over the past 30 years, the supply of raw sewage of domestic, agricultural and industrial origin has ensured the presence of a high concentration of pollutants, including heavy metals. On the basis of the data obtained and in order to restore this sensitive transitional environment, eco compatible interventions are proposed which aim at morphological and hydrologic resettlement, abatement of pollutants on the bottom of the basin, reintroduction of endemic molluscs, together with monitoring the quality of sediments and both sea-lagoon waters and groundwater. PMID- 14749110 TI - Lead contamination and isotope signatures in the urban environment of Hong Kong. AB - The source and the extent of Pb pollution in the urban environment of Hong Kong were investigated at five different urban settings selected on the basis of their annual average daily traffic (AADT) varying from less than 100 to 61,700. In addition, a small distant island without any traffic was selected to establish the possible baseline values. The surface environmental samples studied consisted of street and tunnel dusts, gully sediments, and a limited number of roadside topsoils. The analytical results clearly indicated variable degrees of Pb contamination in these urban settings. However, the level of contamination varied significantly among different types of samples collected at the same location. Pb concentrations of roadside topsoils (79+/-22 micrograms/g) and gully sediments (278+/-88 micrograms/g) were lower than those of the corresponding road dusts (327+/-54 micrograms/g). The Pb isotope compositions in different urban settings varied considerably. The bedrock in the small island had the lowest Pb concentration (12 micrograms/g) but with the highest 206Pb/207Pb ratio (1.2206), whereas the tunnel ceiling dusts with the highest level of Pb (1410 micrograms/g) had the lowest 206Pb/207Pb ratio (1.1062). Despite the significant differences in vehicle types and traffic volumes, and the presence of several different petroleum retailers in Hong Kong, the Pb isotope ratios of road dusts (206Pb/207Pb: 1.1553+/-0.0043, 208Pb/207Pb: 2.4408+/-0.0084) varied within a relatively narrow range among all the five urban sampling sites. On the other hand, the Pb isotopic compositions of gully sediments (206Pb/207Pb: 1.1515+/ 0.0145, 208Pb/207Pb: 2.4322+/-0.0198) varied noticeably within the same setting, but were reasonably comparable across the different study sites. In general, the 206Pb/207Pb ratios of road dusts can be used to estimate the direct contribution from automobile emissions, whereas those of gully sediments might reflect the effects of the mixing of different anthropogenic sources. The Pb isotope signatures in the urban environment of Hong Kong clearly suggested that anthropogenic Pb in the environment originated from Pb ore with a low 206Pb/207Pb ratio (such as the Australian Pb ore and similar sources in Southeast Asia) were significantly different from those of the anthropogenic Pb present in the neighboring Pearl River Delta (PRD) region. PMID- 14749111 TI - Legislation with respect to dioxins in the workplace. AB - Legislation of chemicals is complex and doubts have often been expressed as to its effectiveness, but there are many problems involved in regulating chemicals from synthesis and use through to disposal. Using dioxins as an example of a chemical currently with complicating factors surrounding its control, this paper reviews its occupational legislation, chemistry, formation, toxicity, occurrence, and disposal in industrial situations, and sets this information in context by reference to its environmental occurrence and associated legislation. The role and limitations of legislation to control chemicals like dioxins, which are micropollutants and not intentionally produced, are discussed. Dioxins were chosen as the example because they are widespread, exposure occurs through a variety of routes, and they possess a reputation that ensures public concern. Because dioxins are ubiquitous and are predominantly produced by incineration, whether accidental or deliberate, intake is a total of all exposures from diet, environment and occupation. Occupational exposure is only one factor in this total, but this exposure may push total intake above recommended levels. Although overall exposure is reducing, public concern is rising, and this may be a consequence of greater scientific knowledge. Potential workplace exposures have been evaluated, and a sampling survey was recently undertaken in some of these sectors. The legal options are discussed, and the reasons for some decisions explained. Regulators have an unenviable task, and I believe there is only a fine balance between effective control and over-complicated prescriptive legislation. PMID- 14749112 TI - Worldwide occurrence and effects of antifouling paint booster biocides in the aquatic environment: a review. AB - Organic booster biocides were recently introduced as alternatives to organotin compounds in antifouling products, after restrictions imposed on the use of tributyltin (TBT) in 1987. Replacement products are generally based on copper metal oxides and organic biocides. This ban has led to an increase in alternative coating products containing the above biocides. The most commonly used biocides in antifouling paints are: Irgarol 1051, diuron, Sea-nine 211, dichlofluanid, chlorothalonil, zinc pyrithione, TCMS (2,3,3,6-tetrachloro-4-methylsulfonyl) pyridine, TCMTB [2-(thiocyanomethylthio) benzothiazole], and zineb. Since 1993, several studies have demonstrated the presence of these biocides in European coastal environment as a result of their increased use. More recently, the presence of these biocides was also revealed in waters from Japan, United States, Singapore, Australia and Bermuda. This paper reviews the currently available data on the occurrence of these biocides in the aquatic environment. Some data dealing with the environmental fate, partitioning, behaviour and risk assessment of antifouling paint booster biocides are also reported in order to discuss the detected levels of contamination. PMID- 14749113 TI - Parameters affecting biological phosphate removal from wastewaters. AB - This paper reviews some of the key wastewater composition parameters, which influence the biological removal of phosphate from wastewaters, such as COD content, volatile fatty acid (VFA) content, cation concentration, phosphorus load, pH and food to microorganism ratio. The discussion also focuses on operational parameters affecting successful nutrient removal in wastewater treatment plants, such as temperature, sludge quality, sludge settlement, dissolved oxygen (DO) concentration, anaerobic P-release and secondary P-release. The aim of this review is to compile an updated document for researchers and operators of biological nutrient removal (BNR) systems. In addition, the article will provide a good foundation for readers with no prior knowledge of the process. PMID- 14749114 TI - Metal bioremediation through growing cells. AB - Heavy-metal pollution represents an important environmental problem due to the toxic effects of metals, and their accumulation throughout the food chain leads to serious ecological and health problems. Metal remediation through common physico-chemical techniques is expensive and unsuitable in case of voluminous effluents containing complexing organic matter and low metal contamination. Biotechnological approaches that are designed to cover such niches have, therefore, received great deal of attention in the recent years. Biosorption studies involving low-cost and often dead/pretreated biomass have dominated the literature and, subsequently, extensive reviews focusing on equilibrium and kinetics of metal biosorption have also come up. However, the low binding capacity of biomass for certain recalcitrant metals such as Ni and failure to effectively remove metals from real industrial effluents due to presence of organic or inorganic ligands limit this approach. At times, when pure biosorptive metal removal is not feasible, application of a judicious consortium of growing metal-resistant cells can ensure better removal through a combination of bioprecipitation, biosorption and continuous metabolic uptake of metals after physical adsorption. Such approach may lead to simultaneous removal of toxic metals, organic loads and other inorganic impurities, as well as allow optimization through development of resistant species. However, sensitivity of living cells to extremes of pH or high metal concentration and need to furnish metabolic energy are some of the major constraints of employing growing cells for bioremediation. The efforts to meet such challenges via isolation of metal resistant bacterial/fungal strains and exploitation of organic wastes as carbon substrates have began. Recent studies show that the strains (bacteria, yeast and fungi) isolated from contaminated sites possess excellent capability of metal scavenging. Some bacterial strains possess high tolerance to various metals and may be potential candidates for their simultaneous removal from wastes. Evidently, the stage has already been set for the application of metal-resistant growing microbial cells for metal harvesting. This review focuses on the applicability of growing bacterial/fungal/algal cells for metal removal and the efforts directed towards cell/process development to make this option technically/economically viable for the comprehensive treatment of metal-rich effluents. PMID- 14749115 TI - Carbon trading: time for industry involvement. AB - Carbon trading is no longer just theory. Infant markets already exist and, in 2002, they traded perhaps $10 million worth of emissions allowances. We estimate conservatively that, by 2010, the EU scheme will trade as much as $1 billion in allowances each year. The motor of the carbon markets is a worldwide effort to reduce emissions of greenhouse gases. Its most visible symbol is the Kyoto Protocol of 1997, formally known as the United Nations Framework Convention on Climate Change. Even if Kyoto is not ratified and, despite the US's decision to opt out of the treaty, the genie of greenhouse-gas reduction is out of the bottle. We believe that trading of greenhouse gases will be a real, day-to-day activity by 2010, almost certainly in Europe and probably in Canada and Japan. Carbon trading in these areas will affect the US, whether or not America sets up a programme of its own. The conclusion of this study is that industry should get involved in defining carbon trading-and now-to advance and defend their interests. Interest should be greatest among producers and users of the greenhouse gases other than carbon dioxide, namely methane, nitrous oxide, hydrofluorocarbons, perfluorocarbons and sulphur hexafluoride. PMID- 14749118 TI - Antagonizing XIAP-mediated caspase-3 inhibition. Achilles' heel of cancers? AB - In this issue of Cancer Cell, Schimmer et al. report the identification of small molecule antagonists of XIAP that overcome its inhibition of caspase-3. It was remarkable that the compounds directly induced cell death in tumor cells while having little toxicity on normal cells. This suggests that caspases are already activated in tumor cells, which is different from the caspase activation status in normal mammalian cells. In comparison with Smac peptides targeting XIAP mediated caspase-9 inhibition, which do not directly induce cell death, it appears that liberating downstream caspases rather than upstream caspases may be a preferred strategy for cancer drug discovery. PMID- 14749119 TI - Resistance to antineoplastic therapy. The oncogenic tyrosine kinase-Bcl-x(L) axis. AB - The discovery two decades ago that the Philadelphia chromosome encodes an oncogenic fusion of Bcr and Abl remains among the most important contributions to our understanding of the process of malignant transformation. We now know that Bcr-Abl is one of more than 30 aberrantly activated tyrosine kinases that are expressed in a variety of tumors. Conventional treatment of the tumors in which these proteins are expressed is usually doomed to failure because the activated tyrosine kinases render the tumor cells stubbornly resistant to apoptosis. In this context, it is notable that Zhao and coworkers have uncovered a novel weapon in the resistance armamentarium of these rogue kinases, the suppression of the inactivating deamidation of Bcl-xL (this issue of Cancer Cell). PMID- 14749120 TI - Wnt breakers in colon cancer. AB - Beta-catenin and Tcf4 are the downstream effectors of the Wnt signaling cascade. In colorectal cancer, mutations in Wnt cascade genes such as APC lead to the inappropriate formation of beta-catenin/Tcf4 complexes. Earlier work has predicted that disruption of the beta-catenin/Tcf4 protein-protein interaction could revert the proliferative phenotype of colorectal cancer cells. In this issue of Cancer Cell, Shivdasani and colleagues have explored high-throughput screening of compound libraries in a search for small molecule inhibitors of the Wnt cascade. Ultimately, such inhibitors could become a novel class of smart anticancer drugs. PMID- 14749121 TI - Mouse models of pancreatic cancer: the fur is finally flying! AB - Pancreatic cancer remains one of the most lethal of all human malignancies. Until recently, preclinical studies have been hampered by the absence of mouse models faithfully recapitulating critical elements of the human disease. However, recent months have witnessed a flurry of activity with respect to prospective mouse models. This progress now allows the evaluation of novel strategies for early detection, chemoprevention, and therapy and also provides new insights regarding the potential for differentiated and undifferentiated cell types to act as cells of origin for pancreatic ductal adenocarcinoma. PMID- 14749122 TI - Antiangiogenic therapy and tumor progression. AB - Angiogenesis is necessary for tumor growth (a rationale for antiangiogenic therapy), but hypoxia caused by such a therapy will, in theory, drive tumor progression and metastasis. To reconcile conflicting notions, we discuss that, first, although a shift from normoxia (21% O2) to hypoxia indeed activates cancer cells for aggressive behavior, this may not occur during therapy, because most cancers are not normoxic to start with. Second, only successful antiangiogenic therapy, which is capable of controlling cancer, will select for resistance and progression. After all, in order to occur, therapy-induced tumor progression must be preceded by tumor regression. PMID- 14749123 TI - Focus on epithelial ovarian cancer. PMID- 14749124 TI - Small-molecule antagonists of apoptosis suppressor XIAP exhibit broad antitumor activity. AB - Apoptosis resistance commonly occurs in cancers, preventing activation of Caspase family cell death proteases. XIAP is an endogenous inhibitor of Caspases overexpressed in many cancers. We developed an enzyme derepression assay, based on overcoming XIAP-mediated suppression of Caspase-3, and screened mixture-based combinatorial chemical libraries for compounds that reversed XIAP-mediated inhibition of Caspase-3, identifying a class of polyphenylureas with XIAP inhibitory activity. These compounds, but not inactive structural analogs, stimulated increases in Caspase activity, directly induced apoptosis of many types of tumor cell lines in culture, and sensitized cancer cells to chemotherapeutic drugs. Active compounds also suppressed growth of established tumors in xenograft models in mice, while displaying little toxicity to normal tissues. These findings validate IAPs as targets for cancer drug discovery. PMID- 14749125 TI - An oncogenic tyrosine kinase inhibits DNA repair and DNA-damage-induced Bcl-xL deamidation in T cell transformation. AB - A transgenic mouse model of T cell lymphoma was used to investigate the transforming events mediated by an oncogenic tyrosine kinase in pretumorigenic CD4-CD8- (DN) thymocytes. Parental CD45(-/-) and p56(lck-F505Y) mice do not develop tumors, whereas their CD45(-/-)p56(lck-F505Y) progeny develop T lymphomas. Increased but nononcogenic p56lck kinase activity in p56(lck-F505Y) mice DN thymocytes causes cell-cycle progression, survival, and Bcl-XL upregulation. Additional unique oncogenic signals occur in pretumorigenic CD45(-/ )p56(lck-F505Y) thymocytes in which p56lck kinase activity is 2- to 3-fold higher relative to p56(lck-F505Y): inhibition of DNA repair, inhibition of DNA-damage induced Bcl-XL deamidation, Bax conformational change and mitochondrial translocation, cytochrome c release, and the apoptotic caspase execution cascade. Inhibition of Bcl-XL deamidation may be a critical switch in oncogenic kinase induced T cell transformation. PMID- 14749126 TI - Defective translational control facilitates vesicular stomatitis virus oncolysis. AB - Vesicular stomatitis virus (VSV) exerts potent antitumor activity, although the molecular mechanisms underlying its oncolytic properties remain to be fully clarified. Here, we demonstrate that normally resistant murine embryonic fibroblasts are rendered highly permissive to VSV replication following cellular transformation, a progression that appears to compromise the antiviral effects of interferon (IFN). Subsequent studies revealed normal dsRNA-dependent protein kinase (PKR) activation and phosphorylation of eukaryotic initiation factor 2 (eIF2) alpha. Nevertheless, eIF2B-mediated guanine nucleotide exchange activity downstream of eIF2 was frequently aberrant in transformed cells, neutralizing eIF2alpha phosphorylation and permitting VSV mRNA translation. Thus, defects in translational regulation can cooperate with impaired IFN signaling to facilitate VSV replication, and may represent a common hallmark of tumorigenesis. PMID- 14749127 TI - Synergy in tumor suppression by direct interaction of neutral endopeptidase with PTEN. AB - We show in this study that endogenous NEP and PTEN associate in cells directly through electrostatic interactions between a highly basic residue stretch in the intracellular domain of NEP and the major phosphorylation site in PTEN's tail. NEP binds and engages in higher order complexes both phosphorylated and unphosphorylated PTEN. NEP recruits PTEN to the plasma membrane and enhances its stability and phosphatase activity. As a result, an enzymatically inactive NEP mutant preserves the ability to bind PTEN, inactivates the Akt/PKB kinase, and partially suppresses the growth of PC cells. This study demonstrates a molecular cooperation between NEP and PTEN tumor suppressors in which NEP constitutively recruits and activates PTEN to inhibit the PI3K/Akt oncogenic pathway. PMID- 14749128 TI - Regulation of tumor angiogenesis by integrin-linked kinase (ILK). AB - We show that integrin-linked kinase (ILK) stimulates the expression of VEGF by stimulating HIF-1alpha protein expression in a PKB/Akt- and mTOR/FRAP-dependent manner. In human prostate cancer cells, knockdown of ILK expression with siRNA, or inhibition of ILK activity, results in significant inhibition of HIF-1alpha and VEGF expression. In endothelial cells, VEGF stimulates ILK activity, and inhibition of ILK expression or activity results in the inhibition of VEGF mediated endothelial cell migration, capillary formation in vitro, and angiogenesis in vivo. Inhibition of ILK activity also inhibits prostate tumor angiogenesis and suppresses tumor growth. These data demonstrate an important and essential role of ILK in two key aspects of tumor angiogenesis: VEGF expression by tumor cells and VEGF-stimulated blood vessel formation. PMID- 14749129 TI - Small-molecule antagonists of the oncogenic Tcf/beta-catenin protein complex. AB - Key molecular lesions in colorectal and other cancers cause beta-catenin dependent transactivation of T cell factor (Tcf)-dependent genes. Disruption of this signal represents an opportunity for rational cancer therapy. To identify compounds that inhibit association between Tcf4 and beta-catenin, we screened libraries of natural compounds in a high-throughput assay for immunoenzymatic detection of the protein-protein interaction. Selected compounds disrupt Tcf/beta catenin complexes in several independent in vitro assays and potently antagonize cellular effects of beta-catenin-dependent activities, including reporter gene activation, c-myc or cyclin D1 expression, cell proliferation, and duplication of the Xenopus embryonic dorsal axis. These compounds thus meet predicted criteria for disrupting Tcf/beta-catenin complexes and define a general standard to establish mechanism-based activity of small molecule inhibitors of this pathogenic protein-protein interaction. PMID- 14749131 TI - Elevated sex-hormone binding globulin in elderly women with Alzheimer's disease. AB - BACKGROUND: Hormone levels change significantly with increasing age. These changes may be related to, or be associated with, the emergence of age-related diseases, such as Alzheimer's disease (AD). METHODS: Five hundred and seventy-six women over the age of 65 were studied from the Washington Heights-Inwood Columbia Aging Project (WHICAP). These women were selected from a group of healthy Medicare beneficiaries that were aged 65 and older living in the geographically defined area of northern Manhattan in New York City. Serum levels of estrone (E1), estradiol (E2), total testosterone (TT), dehydroepiandosterone (DHEA), luteinizing hormone (LH), follicle stimulating hormone (FSH), and sex-hormone binding globulin (SHBG) were measured. RESULTS: Significant differences were found between patients with AD and controls only in the level of SHBG, which was 20% higher in patients compared to controls (68.5nmol/l versus 54.7nmol/l, P<0.001). We also estimated levels of total E2 because after menopause, E2 is largely derived from E1. AD patients had significantly lower levels of estimated E2 (AD 0.46 versus controls 0.49, P<0.01). Differences remained significant after adjusting for age, ethnic group, education, and body mass index (BMI). CONCLUSIONS: A marked increase in SHBG levels was found in AD patients. SHBG normally responds to circulating testosterone and estrogen, therefore, elevated SHBG suggests an abnormal increase in its production and regulation. Further work is needed to clarify the cause and consequences of this observation. PMID- 14749132 TI - Glutamate transporters in platelets: EAAT1 decrease in aging and in Alzheimer's disease. AB - Platelets release glutamate upon activation and are an important clearance system of the amino acid from blood, through high-affinity glutamate uptake, similar to that described in brain synaptosomes. Since platelet glutamate uptake is decreased in neurodegenerative disorders, we performed a morphological and molecular characterization of platelet glutamate transporters. The three major brain glutamate transporters EAAT1, EAAT2 and EAAT3 are expressed in platelets, with similar molecular weight, although at lower density than brain. A Na(+) dependent-high-affinity glutamate uptake was competitively inhibited by known inhibitors but not by dihydrokainic acid, suggesting platelet EAAT2 does not play a major role in glutamate uptake at physiological conditions. We observed decreased glutamate uptake V(max), without modification of transporter affinity, in aging, which could be linked to the selective decrease of EAAT1 expression and mRNA. Moreover, in AD patients we found a further EAAT1 reduction compared to age matched controls, which could explain the decrease of platelet uptake previously described. Platelet glutamate transporters may be used as peripheral markers to investigate the role of glutamate in patients with neuropsychiatric disorders. PMID- 14749133 TI - Obesity enhances verbal memory in postmenopausal women with Down syndrome. AB - Several lines of evidence suggest that the loss of estrogen after menopause may play a role in cognitive declines associated with Alzheimer's disease (AD). In postmenopausal women, the principal source of estrogen is estrone, which is influenced by body mass index (BMI). Increased BMI in postmenopausal women is associated with higher levels of serum estradiol and estrone. We hypothesized that obesity could have a beneficial effect on cognition with advancing age. We compared the performance of healthy nondemented obese and non-obese women with Down syndrome (DS) on a broad spectrum of cognitive tests. Estrone levels were 66.9% higher in obese than in non-obese postmenopausal women, and 136% higher in obese than in non-obese premenopausal women. Obese postmenopausal women performed significantly better than non-obese women on measures of verbal memory and on an omnibus test of neuropsychological function, but did not differ significantly in verbal fluency, language, praxis or visuospatial functioning. Among premenopausal women, there was no difference in cognitive function between obese and non-obese women. Our results support the hypothesis that higher endogenous estrogen levels after menopause are associated with better performance on verbal memory. PMID- 14749134 TI - Serotonin modulation of cerebral glucose metabolism in normal aging. AB - Age-related alterations in serotonin function may increase the vulnerability to psychiatric and neurodegenerative disorders in late life. The neuroendocrine and cerebral metabolic response to the acute administration of the selective serotonin reuptake inhibitor, citalopram (40mg, IV), was measured in 17 normal control subjects using positron emission tomography (PET) to evaluate changes in serotonin function with normal aging. The citalopram-induced change in cerebral metabolism was positively correlated with age in the right precuneus, right paracentral lobule, and left middle temporal gyrus and negatively correlated with age in the left anterior cingulate gyrus, right inferior and middle frontal gyri, right insula, and right inferior parietal lobule. The positive correlations in mainly posterior brain regions indicate that normal aging is associated with an increase in metabolism after citalopram administration, whereas the negative correlations in mainly anterior brain regions indicate that normal aging is associated with a greater decrease in metabolism. These results suggest different compensatory processes in anterior compared to posterior brain regions secondary to the age-related loss of serotonin innervation. PMID- 14749135 TI - Morphological changes in aging brain structures are differentially affected by time-linked environmental influences despite strong genetic stability. AB - This longitudinal study used the full twin model to estimate change and stability of genetic contributions to morphology of two brain structures, the corpus callosum and lateral ventricles. The 142 subjects were 34 monozygotic (MZ) and 37 dizygotic (DZ) elderly male twin pairs from the National Heart, Lung, and Blood Institute (NHLBI) Twin Study who underwent brain magnetic resonance imaging twice, separated by a 4-year interval. Genetic factors accounted for a substantial portion of individual differences in the size of the corpus callosum and its substructures and of lateral ventricular size. Longitudinal genetic analyses revealed no significant change in the heritability of these structures and no evidence for new genetic variance at Time 2 not present at Time 1. However, both the callosal and ventricular measures showed evidence for new environmental variance at Time 2 not present at Time 1. Confirming a previously posed hypothesis, the phenotypic correlation between absolute change in height of the corpus callosum and absolute change in ventricular volume was significant. Bivariate genetic analysis estimated a significant genetic correlation between the changes in these two structures and the genetic variance in the change of callosal height was entirely due to genes involved in the expansion of ventricles. Genetic stability was present even in old age when brain and other morphological changes can be rapid and highly variable across individuals, inconsistent with an hypothesis that random DNA damage is the cause of aging. PMID- 14749136 TI - Effects of age and sex on volumes of the thalamus, pons, and cortex. AB - Volumes of thalamus, pons, cortical gray matter, and white matter were derived from MR brain images of healthy men and women spanning the adult age range in order to delineate patterns of aging and to compare age and sex effects in thalamus and pons with such effects in cortical gray and white matter volumes. Men had larger intracranial volume (ICV) than women, but ICV did not correlate with age in either sex. Thalamic, pontine, and cortical white matter volumes did not differ between men and women once ICV differences were taken into account, but men had more cortical gray matter than women even after accounting for ICV. Volumes of pons and thalamus were associated, independent of ICV, in women but not in men. Thalamic volume declined linearly with age at a similar rate in both men and women, whereas cortical gray matter volume declined more steeply with age in men than women. Both pontine and cortical white matter volumes remained stable across the age span in both men and women. PMID- 14749137 TI - Age-related decline in striatal volume in rhesus monkeys: assessment of long-term calorie restriction. AB - Using magnetic resonance imaging (MRI), we measured striatal volume in 22 male rhesus monkeys undergoing calorie restriction (CR) for 11-13 years and 38 monkeys who were fed ad libitum (CON). CR delays the onset of many age-related processes, and this study tested whether it would alter the age-related decline in striatal volume. The CON and CR groups were sub-divided into middle age (less than 24 years old) and old age groups. Contrary to expectation, volumes of the putamen (not the caudate nucleus) were larger bilaterally in the CON than in the CR group both at middle age and senescence. Regression analysis (region volume versus age) indicated bilateral age-related declines in putamen and caudate nucleus volumes in the old CON monkeys, but only for the putamen in the old CR monkeys. Because tests for slopes found no differences between the groups, the data do not establish an effect of CR. Further study, involving sequential imaging, is warranted in order to clarify the possible effects of CR on age-related changes in striatal volume. PMID- 14749138 TI - Changes in the expression of the NR2B subunit during aging in macaque monkeys. AB - Humans, non-human primates and rodents show declines in spatial memory abilities with increased age. Some of these declines in mice are related to changes in the expression of the epsilon2 (epsilon2) (NR2B) subunit of the N-methyl-D-aspartate receptor. The purpose of this study was to determine whether primates show changes during aging in the mRNA expression of the NR2B subunit. In situ hybridization was performed on tissue sections from three different ages of Rhesus monkeys (Macaca mulatta; 6-8, 10-12, and 24-26 years). There was a significant decrease in the mRNA expression of the NR2B subunit overall in the prefrontal cortex and in the caudate nucleus between young and old monkeys. There were no significant changes in NR2B mRNA expression in the hippocampus or the parahippocampal gyrus. The results in the prefrontal cortex, caudate and hippocampus were similar to those seen previously in C57BL/6 mice during aging, which suggests that mice may be useful as a model for primates to further examine the age-related changes in the expression of the NR2B subunit of the NMDA receptor in several important regions of the brain. PMID- 14749139 TI - Reduced cortical noradrenergic neurotransmission is associated with increased neophobia and impaired spatial memory in aged rats. AB - In the present study, young (5-month-old (mo)) and aged (24 mo) adult male Fischer-344 (F344) rats were assigned to experimental groups based upon their performance of a reference memory task in the Morris water maze and reactivity to a novel palatable taste in a gustatory neophobia task. Levels of norepinephrine (NE) and its metabolite 3-methoxy-4-hydroxy-phenylglycol (MHPG) were assayed via high performance liquid chromatography (HPLC) in brain regions associated with the locus coeruleus (LC)-hippocampus-cortex system and A1/A2-hypothalamic system. Binding of ligands specific for alpha-1, alpha-2, beta-1, and beta-2 receptors was assessed in hippocampus and cortex with receptor autoradiography. Impaired acquisition and retention of the water maze task and gustatory neophobia in aged rats was primarily associated with decreased NE activity in cingulate cortex (CC) as indicated by a significant reduction in the MHPG/NE ratio coupled with increased NE content. No significant changes in adrenergic receptor binding were detected in any region sampled. The results suggest that an aging-related reduction in cortical NE neurotransmission is associated with the expression of increased neophobia and deficits in spatial learning and memory performance occurring with advanced age in rats. PMID- 14749140 TI - Effect of age on the radial arm water maze-a test of spatial learning and memory. AB - Aged rats show decrements in performance on cognitive tasks that require the use of spatial learning and memory. We used the 8-arm radial water maze (RAWM) to measure spatial learning as a function of age in young (6 months) and old (21 months) male F344 rats. Rats were placed in the RAWM in different start arms with the same goal arm for 3 days (five trials/day); the goal arm was changed on day 4. Old rats demonstrated spatial impairment as evidenced by increased latencies to find the hidden platform on day 4. Old rats made significantly more errors, both reference and working memory errors, than young rats on all days. It is likely that the old rats utilized non-spatial strategies to solve the task, and therefore were impaired in learning a new platform location. The RAWM is a reliable, sensitive, and powerful additional test to assess age-related spatial learning and memory deficits, combining the advantages of the Morris water maze and the radial arm maze while minimizing the disadvantages. PMID- 14749142 TI - Abstracts of the 2nd Meeting of the Alzheimer's Imaging Consortium. Stockholm, Sweden, July 20, 2002. PMID- 14749141 TI - The effect of age on prepro-orexin gene expression and contents of orexin A and B in the rat brain. AB - Orexin A and B (hypocretin 1 and 2) are hypothalamic peptides, which are synthesized in the lateral hypothalamus. Orexins participate in the regulation energy balance, food intake, vigilance and several endocrine and autonomic functions. The widespread projections of the orexin neurons suggest that they may have a role in coordination of different brain activities. The effects of ageing on the orexin system have not been studied previously. Prepro-orexin gene expression in the lateral hypothalamus, and the contents of orexin A and B peptides in the lateral hypothalamus and hypothalamus were measured in young, middle-aged and old (3, 12 and 24 months) rats. In the course of ageing, the expression of the prepro-orexin gene and the levels of orexin A and B decreased; the main decrease occurred by 12 months. Sleep deprivation for 6h increased slightly the expression of prepro-orexin gene in young rats. Deterioration of the orexin system may play a role in the phenomenon associated with aging, e.g. decreased consolidation of vigilance states, endocrine changes and dysfunctions of autonomic nervous system. PMID- 14749143 TI - Metformin hydrochloride in the treatment of type 2 diabetes mellitus: a clinical review with a focus on dual therapy. AB - BACKGROUND: Type 2 diabetes mellitus typically involves abnormal beta-cell function that results in relative insulin deficiency, insulin resistance accompanied by decreased glucose transport into muscle and fat cells, and increased hepatic glucose output, all of which contribute to hyperglycemia. OBJECTIVE: This review examines the pharmacology, pharmacokinetics, drug interaction potential, adverse effects, and dosing guidelines for metformin hydrochloride, a biguanide agent for the treatment of type 2 diabetes. Clinical trial data are reviewed, including efficacy and tolerability information, with a focus on studies of dual metformin therapy (metformin plus another oral agent or insulin) published from 1998 to the present. Pharmacoeconomic considerations are also discussed. METHODS: Primary research and review articles were identified through a search of MEDLINE (1966-May 2003) and International Pharmaceutical Abstracts (1970-May 2003) using the terms metformin and/or Glucophage. Web of Science (1995-May 2003) was used to search for additional abstracts. The package inserts for metformin and metformin combination products were consulted. All identified articles and abstracts were assessed for relevance, and all relevant information was included. Priority was given to the primary medical literature and clinical trial reports. RESULTS: Metformin is the only currently available oral antidiabetic/hypoglycemic agent that acts predominantly by inhibiting hepatic glucose release. Because patients with type 2 diabetes often have excess hepatic glucose output, use of metformin is effective in lowering glycosylated hemoglobin (HbA1c) by 1 to 2 percentage points when used as monotherapy or in combination with other blood glucose-lowering agents or insulin. Other metabolic variables (eg, dyslipidemia, fibrinolysis) may be improved with the use of metformin. Body weight is often maintained or slightly reduced from baseline. Metformin is well tolerated and is associated with few clinically deleterious adverse events. The most important and potentially life-threatening adverse event associated with its use is lactic acidosis, which occurs very rarely. CONCLUSIONS: Metformin has multiple benefits in patients with type 2 diabetes. It can effectively lower HbA1c values, positively affect lipid profiles, and improve vascular and hemodynamic indices. Adverse effects are generally tolerable and self-limiting. The availability of products combining metformin with a sulfonylurea or rosiglitazone has expanded the array of therapies for the management of type 2 diabetes. PMID- 14749144 TI - Estratest and Estratest HS (esterified estrogens and methyltestosterone) therapy: a summary of safety surveillance data, January 1989 to August 2002. AB - BACKGROUND: The use of hormone-replacement therapy (HRT) to treat menopausal symptoms has been influenced over the years by various safety concerns. These concerns include endometrial cancer, breast cancer, and cardiovascular disease, and have altered how HRT is prescribed. Evaluating postmarketing surveillance data for a product can help pharmaceutical manufactures and health care providers detect early safety signals that may call for further investigation of the product for safety risks. OBJECTIVE: This review summarizes the safety surveillance data for Estratest and Estratest HS from January 1989 to August 2002. METHODS: All adverse-event (AE) data reported to Solvay Pharmaceuticals, Inc., on this brand from January 1989 to August 2002 were accessed from a database system that uses a comprehensive software package for reporting and tracking clinical and postmarketing AEs. RESULTS: Exposure to the Estratest brand during the 13-year assessment period is estimated at >3.0 million patient-years. A total of 1372 unique case reports containing 2556 AEs were found. Assessment of the 43 (3.1%) serious AE cases reported did not generate any signals that might raise concern on the part of the medical community or consumers. Nonserious events comprising >4% of total AEs were all labeled events and included alopecia (8.8%), acne (5.6%), and hirsutism (4.5%). CONCLUSIONS: The relatively small number of serious AE reports compared with the significant patient exposure did not generate any signals that might raise concern on the part of the medical community or consumers. The safety profile suggests that continued use at the lowest effective dose is acceptable in menopausal women whose symptoms are not improved by estrogen alone. PMID- 14749145 TI - Losartan and other angiotensin II antagonists for nephropathy in type 2 diabetes mellitus: a review of the clinical trial evidence. AB - BACKGROUND: End-stage renal disease (ESRD) in patients with type 2 diabetes mellitus (DM) is associated with a bleak prognosis. The life span of patients with DM who have undergone renal transplantation or who are undergoing dialysis is up to 30% shorter than that of individuals in the general population. Preventing or delaying the progression of renal disease from microalbuminuria to nephropathy, and ultimately, to ESRD is thus a crucial goal of DM management. OBJECTIVE: This article reviews the growing worldwide problem of type 2 DM and ESRD, the renoprotective benefits of angiotensin II (AII) antagonists (AIIAs) such as losartan in patients with or without type 1 or 2 DM, potential mechanisms of renoprotection of AIIAs beyond blood pressure (BP) control, and the clinical practice implications of available megatrials. METHODS: Articles included in this review were identified using a MEDLINE search for English-language studies published between 1990 and 2003 and included the search terms diabetic nephropathy, type 2 diabetes mellitus, microalbuminuria, proteinuria, angiotensin II antagonists, angiotensin-converting enzyme inhibitors, and cardiovascular disease. Articles describing major clinical trials, new data, or new mechanisms pertinent to the management of type 2 DM were selected for review. RESULTS: Currently, AIIAs such as losartan represent the only evidence-based treatment strategy for patients with type 2 DM and proteinuria. The Irbesartan in Patients with Type 2 Diabetes and Microalbuminuria (IRMA-2) study, the Reduction of End Points in Non-Insulin Dependent Diabetes Mellitus with the Angiotensin II Antagonist Losartan (RENAAL) study, and the Irbesartan Type 2 Diabetic Nephropathy Trial (IDNT) indicate that AIIAs postpone the progression of type 2 diabetic renal disease at all stages, ranging from microproteinuria to overt nephropathy and ESRD, RENAAL showed that losartan improves renal outcomes in patients with type 2 DM and nephropathy over and above that attributable to BP control alone. The renoprotective effect of losartan corresponded to an average delay of 2 years in the need for dialysis or kidney transplantation. CONCLUSIONS: AIIAs such as losartan should perhaps be considered mandatory therapy in patients with diabetic nephropathy and should complement existing management strategies, such as reduced dietary protein intake, strict blood glucose control, and standard antihypertensive therapy. Collectively, these measures should improve survival and quality of life and reduce the health care burden of managing patients with diabetic nephropathy. PMID- 14749146 TI - Atomoxetine hydrochloride for the treatment of attention-deficit/hyperactivity disorder. AB - BACKGROUND: Attention-deficit/hyperactivity disorder (ADHD) occurs in approximately 3% to 10% of the pediatric population. Most of the drugs typically used to treat ADHD are stimulants, which, because of their addictive properties and potential for abuse, are controlled substances. Although these drugs are the mainstay of treatment for ADHD, nearly one third of patients may not respond to or be able to tolerate them. Atomoxetine hydrochloride, a nonstimulant approved by the US Food and Drug Administration for the treatment of ADHD, may provide an alternative to the use of stimulants. OBJECTIVE: The goal of this review was to describe the chemistry, mechanism of action, pharmacokinetics, drug interactions, and efficacy and safety profiles of atomoxetine in pediatric and adult patients with ADHD, as well as relevant pharmacoeconomic considerations. METHODS: Relevant publications were identified through a search of the English-language literature indexed on PreMEDLINE and MEDLINE (1966-May 2003) using the search terms atomoxetine, tomoxetine, and LY139603. These terms were also applied to the Google search engine. All articles were reviewed for suitability for inclusion. The manufacturer of atomoxetine provided both published and unpublished data. RESULTS: In the data reviewed, atomoxetine was more efficacious than placebo in patients with ADHD (P<0.05 to P<0.01). Therapeutic doses ranged from 45 mg or placebo (P<0.05). These results add support to the hypothesis that atomoxetine may not cause the increase in dopamine concentrations in the nucleus accumbens that is associated with pleasurable effects and abuse potential. PMID- 14749147 TI - Adefovir dipivoxil: a new antiviral agent for the treatment of hepatitis B virus infection. AB - BACKGROUND: Hepatitis B virus (HBV) infection is a major problem worldwide and an important cause of chronic liver disease and cirrhosis. Currently available treatments include interferon alfa-2b, lamivudine, and adefovir dipivoxil. Adefovir dipivoxil is an acyclic nucleotide analogue that was developed in part to improve on the limitations of earlier therapies. OBJECTIVE: This article is a review of available data on the clinical pharmacology, virology, efficacy, tolerability, and clinical use of adefovir dipivoxil. METHODS: A search of the English-language literature indexed on MEDLINE from 1966 to July 2003 was performed using the terms adefovir, PMEA, and Bis-POM PMEA. Pertinent abstracts from scientific meetings on infectious diseases and hepatology were also included. The manufacturer of adefovir dipivoxil provided additional information. These materials were supplemented by US Food and Drug Administration briefing documents and other unpublished materials. In vitro and preclinical studies were included in the review, as were Phase II and III clinical trials. RESULTS: In vitro, adefovir dipivoxil concentrations exceed those necessary to inhibit both wild-type and lamivudine-resistant isolates of HBV. In clinical trials, adefovir dipivoxil was clinically and virologically effective in patients in whom lamivudine therapy had failed due to the presence of lamivudine-resistant HBV. The drug was generally well tolerated. The risk of nephrotoxicity, the most notable adverse effect of adefovir dipivoxil at previously used higher doses, has been substantially reduced at the currently recommended dosage of 10 mg/d. CONCLUSION: Based on the data reviewed adefovir dipivoxil is an effective and well-tolerated alternative for the treatment of HBV infection, including disease that is lamivudine resistant. PMID- 14749148 TI - Randomized, double-blind, placebo-controlled, dosed-finding study of the antiemetic effects and tolerability of ramosetron in adults undergoing middle ear surgery. AB - BACKGROUND: Ramosetron is a selective serotonin receptor antagonist (SSRA) that is approved for the treatment of emetic symptoms induced by cytotoxic drugs in Japan. We have reported that ramosetron 0.3 mg had comparable efficacy to granisetron 3 mg, another SSRA, in preventing emetic symptoms in adults in the first 48 hours after the start of anesthesia for middle ear surgery. Although it has been shown that a high dose of ramosetron can cause adverse effects (AEs), such as severe headache, the minimal effective dose of ramosetron is unknown. OBJECTIVE: The aim of this study was to determine the minimum effective and tolerable dose of ramosetron needed to prevent postoperative emetic symptoms in adult patients undergoing middle ear surgery. METHODS: This randomized, double blind, placebo-controlled, dose-finding study was conducted at the Department of Anesthesiology, Toride Kyodo General Hospital (Toride, Japan). Patients aged > or =20 years scheduled for middle ear surgery were randomized to receive either placebo or ramosetron at 1 of 3 doses (0.15, 0.3, or 0.6 mg), regardless of body weight, i.v. immediately before anesthesia induction. Emetic symptoms (nausea, retching, or vomiting) occurring from 0 to <24 and 24 to 48 hours after the start of anesthesia were recorded. Other AEs also were assessed. RESULTS: A total of 100 patients (55 women, 45 men; mean [SD] age, 44 [12] years; mean [SD] body weight, 56 [8] kg; mean [SD] height, 159 [8] cm) were enrolled. Each treatment group comprised 25 patients. The treatment groups were comparable with regard to demographic characteristics and type of surgery After the second 24 hour postanesthesia period, significantly more patients in the ramosetron 0.3-mg and 0.6-mg groups were emesis free than in the placebo group (both P<0.001). The number of emesis-free patients in the ramosetron 0.15-mg group and the placebo group were similar after both study periods. No significant difference in antiemetic efficacy was found between the ramosetron 0.3-mg and 0.6-mg groups. No relationship between body weight and the efficacy of ramosetron was observed. The incidence of AEs was similar in all 4 groups. CONCLUSIONS: Ramosetron 0.3 mg, regardless of body weight, was more effective than either ramosetron 0.15 mg or placebo and as effective as ramosetron 0.6 mg for the prevention of emetic symptoms in the first 48 hours after the start of anesthesia in this selected population of adult patients who underwent middle ear surgery. PMID- 14749149 TI - Addition of biphasic insulin aspart 30 to rosiglitazone in type 2 diabetes mellitus that is poorly controlled with glibenclamide monotherapy. AB - BACKGROUND: The incidence of type 2 diabetes mellitus (DM) is rapidly increasing worldwide. Results from large-scale studies show that tight blood glucose (BG) control improves the outcome of patients with type 2 DM. OBJECTIVE: This trial assessed the short-term efficacy and tolerability of adding a thiazolidinedione (rosiglitazone [ROS]) to existing sulfonylurea (SU) therapy (glibenclamide) compared with switching to combination treatment with a premixed insulin (biphasic insulin aspart 30 [BIAsp 30], a rapid-acting insulin analog) and the thiazolidinedione in a select group of patients with type 2 DM whose metabolic control was inadequate with SU monotherapy. METHODS: In this 6-week, multicenter, open-label, parallel-group trial, patients with type 2 DM whose BG level was not adequately controlled with glibenclamide monotherapy (glycosylated hemoglobin [HbA1c] 8%-13%) were randomized either to replace glibenclamide with BIAsp 30 (individually titrated dosages starting with 6-8 U BID) plus rosiglitazone (4 mg once daily) (BIAsp 30 + ROS group) or to add rosiglitazone (4 mg once daily) to their pretrial doses of glibenclamide (GLIB + ROS group). Patients measured their BG levels immediately before each of the 3 main meals, 90 minutes after the start of each meal, and at bedtime, and mean BG levels were calculated at weeks 0 (baseline), 1, 2, 4, 6, and at 2-week follow-up (week 8). The primary end point was change in mean daily BG level during treatment. Secondary end points included preprandial, postprandial, and bedtime BG levels, serum fructosamine level HbA, and fasting BG level, which were measured at each study visit. Tolerability was assessed using hematologic and biochemical parameters, vital signs, and physical examination. RESULTS: Forty-nine patients (32 men, 17 women; mean [SD] age, 59.1 [8.9] years; mean [SD] body mass index, 27.7 [3.7] kg/m2) participated in the study. A significant difference was found between treatments in the change in mean daily BG level from baseline to week 6 (P=0.01). After the 6-week treatment period, change in mean serum fructosamine level was significantly greater for BIAsp 30 + ROS compared with GLIB + ROS (P=0.02). HbA1c decreased in both treatment groups from baseline to study end, but the difference between groups was nonsignificant. The changes in fasting BG from baseline to study end also were nonsignificant between groups. Both combinations were well tolerated. CONCLUSIONS: This short-term study in patients with type 2 DM whose BG level was poorly controlled with glibenclamide monotherapy suggests that switching to a combination of BIAsp 30 + ROS was efficacious and well tolerated and provided an alternative to adding rosiglitazone to existing glibenclamide treatment. The study also suggests that BIAsp 30 may be associated with greater improvements in short-term metabolic control. PMID- 14749150 TI - Oral versus intramuscular cobalamin treatment in megaloblastic anemia: a single center, prospective, randomized, open-label study. AB - BACKGROUND: Cobalamin (vitamin B12) deficiency, the most common cause of megaloblastic anemia, is treated with intramuscular (IM) cobalamin. It has been suggested by some investigators that oral (p.o.) cobalamin treatment may be as effective in the treatment of this condition, with the advantages of ease of administration and lower cost. OBJECTIVE: This study assessed the effects and cost of p.o. versus i.m. cobalamin treatment in patients with megaloblastic anemia due to cobalamin deficiency. METHODS: This was a 90-day, prospective, randomized, open-label study conducted at the Division of Hematology, Department of Internal Medicine, Adnan Menderes University Research and Practice Hospital (Aydin, Turkey). Patients aged > or =16 years with megaloblastic anemia due to cobalamin deficiency were randomized to receive 1000-microg cobalamin p.o. once daily for 10 days (p.o. group) or 1000-microg cobalamin i.m. once daily for 10 days (i.m. group). After 10 days, both treatments were administered once a week for 4 weeks, and after that, once a month for life. Patients were assessed for the presence of reticulocytosis between treatment days 5 and 10 until it was detected. Therapeutic effectiveness was assessed by measuring hematologic parameters on days 0, 10, 30, and 90 and serum vitamin B12 concentration on days 0 and 90. The Mini-Mental State Examination was used before and after the B12 therapy for cognitive function assessment and 125-Hz diapozone was used for vibration threshold testing. Neurologic sensory assessment, including soft-touch and pinprick examinations, was used to identify neuropathy at baseline and study end. Tolerability was assessed using laboratory tests and patient interview. Cost was assessed using the cost of the study drug and of the injection. RESULTS: Sixty patients completed the study 26 in the p.o. group (16 men, 10 women; mean [SD] age, 60 [15] years) and 34 in the i.m. group (17 men, 17 women; mean [SD] age, 64 [10] years). Reticulocytosis was observed in all patients. In the p.o. group, at days 30 and 90, all hematologic parameters changed significantly versus day 0 (mean hemoglobin levels increased [both P<0.001]; mean corpuscular volume decreased [both P<0.001]; mean white blood cell count increased [day 30, P<0.01; day 90, P<0.001]; and mean platelet count increased [both P<0.001]). The mean serum vitamin B12 concentration increased significantly from day 0 to 90 (P<0.001). These hematologic parameters and the recovery patterns were similar between the 2 groups. Neurologic findings included sensitive peripheral neuropathy in 9 patients (15.0%), alteration of cognitive function (loss of memory, impaired concentration) in 7 patients (11.7%), and loss of sense of vibration in 5 patients (8.3%). Neurologic improvement was detected in 7 of 9 patients (77.8%) in the p.o. group and 9 of 12 patients (75.0%) in the i.m. group at day 30. CONCLUSIONS: In this study of patients with megaloblastic anemia due to cobalamin deficiency, p.o. cobalamin treatment was as effective as i.m. cobalamin treatment. P.o. treatment also was better tolerated and less expensive compared with IM treatment. However, because of the small sample size and the short term of this study, further long-term studies are needed to determine the efficacy of p.o. cobalamin treatment. PMID- 14749151 TI - Results of a prospective, randomized, double-blind, placebo-controlled, dose ranging trial to determine the effective dose of ramosetron for the prevention of vomiting after tonsillectomy in children. AB - BACKGROUND: Postoperative vomiting (POV) is an important adverse effect of anesthesia and surgery, and children undergoing tonsillectomy may be particularly at risk. OBJECTIVE: The aim of this study was to determine the effective dose of ramosetron, a 5-hydroxytryptamine type 3 receptor antagonist, for prophylaxis of severe POV (> or =2 episodes) in children undergoing general anesthesia for tonsillectomy. METHODS: Standard general anesthetic technique and postoperative analgesia were used in this prospective, randomized, double-blind, placebo controlled, dose-ranging trial of pediatric patients. Patients who had experienced POV, had taken an antiemetic medication within 24 hours before surgery of had a history of motion sickness were excluded. Only patients aged 4 to 10 years were included, because of their ability to answer questions. Patients received a single administration of either i.v. placebo or i.v. ramosetron at 3, 6, or 12 microg/kg immediately after the end of surgery. During the first 48 hours after anesthesia, episodes of retching, vomiting, and adverse events were recorded by nursing staff blinded to treatment assignment. RESULTS: Eighty children (20 in each group)--42 girls and 38 boys--were enrolled. There were no differences in patient demographic characteristics among the treatment groups. The rates of complete response (no vomiting, retching, or need for another antiemetic medication) from 0 to 24 hours after anesthesia were 35% (7/20) with ramosetron 3 microg/kg, 90% (18/20) with ramosetron 6 micro/kg, and 90% (18/20) with ramosetron 12 microg/kg compared with placebo (30% [620], P=NS, P=0.001, and P=0.001 vs placebo, respectively); the corresponding rates from 24 to 48 hours after anesthesia were 35% (7/20), 90% (18/20), and 95% (19/20) compared with placebo (35% [7/20]; P=NS, P=0.001, and P=0.001 vs placebo, respectively). No difference in antiemetic efficacy was observed between ramosetron 6 and 12 microg/kg. No clinically serious adverse events attributable to the study drug were observed in any group. CONCLUSIONS: In the pediatric population studied, ramosetron 6 microg/kg was effective for the prevention of vomiting after tonsillectomy from 0 to 48 hours after anesthesia. Increasing the dose to 12 microg/kg did not appear to provide further benefit. PMID- 14749153 TI - Retrospective, long-term follow-up study of the effect of a three-tier prescription drug copayment system on pharmaceutical and other medical utilization and costs. AB - BACKGROUND: Previous research has suggested that 3-tier prescription drug copayment systems produce drug cost savings without affecting the use of other medical services during the first 12 months after implementation. Assessment of such systems with a longer follow-up period has been needed. OBJECTIVE: This study examined the effect of a 3-tier copayment system on pharmaceutical and medical utilization and cost for 30 months after implementation in a population of commercially insured, preferred-provider organization members. METHODS: This was a quasi-experimental, pre-post with comparison group design that gathered data retrospectively from the claims database of a preferred-provider organization in the Midwestern United States. The intervention group comprised members whose employer switched from a 2-tier (generic/brand copayment) plan to a 3-tier (generic/formulary/nonformulary) plan. The comparison group comprised members whose employer retained the 2-tier plan. Employers did not offer a choice between the 2- and 3-tier plans. Outcome measures included total drug cost; net insurer cost (drug cost minus copayment); number of prescription claims; numbers of office visits, inpatient hospitalizations, and emergency department visits; and rates of continuation with chronic medication therapy. RESULTS: Relative to the comparison group (n=4132), the intervention group (n=3577) showed reduced growth in net cost and lower utilization of third-tier (nonformulary) medications (P<0.001 and P<0.01, respectively). The intervention and comparison groups did not differ significantly with respect to numbers of office visits, emergency department visits, or inpatient hospitalizations. Medication continuation rates were lower for the intervention than the comparison group at 6 months for oral contraceptives (P<0.05), but chronic medication therapy continuation rates did not differ significantly at any other time point or for estrogens, antihypertensives, or antihyperlipidemics. CONCLUSION: In the population studied, previous research findings were confirmed over a longer time period. PMID- 14749155 TI - Cost analysis of switching from i.v. vancomycin to p.o. linezolid for the management of methicillin-resistant Staphylococcus species. AB - BACKGROUND: Infections with methicillin-resistant Staphylococcus species (MRSS) are associated with higher treatment costs than infections with methicillin sensitive Staphylococcus species in the United States--partly because of an increased length of hospital stay (LOS). OBJECTIVE: This study used pharmacoeconomic modeling to evaluate the costs and outcomes associated with the use of i.v. vancomycin compared with p.o. linezolid in the treatment of MRSS infected patients. METHODS: A retrospective chart review was used to determine the number of cases with confirmed or presumed MRSS infections treated with i.v. vancomycin during calendar-year 2000 at the Veterans Affairs Greater Los Angeles Healthcare System inpatient facility. Patients who were eligible for a switch to p.o. linezolid with or without early discharge to home were identified. Cost differences associated with conversion from i.v. to p.o. therapy (compared with continued i.v. therapy) were estimated based on a mean decreased LOS and a decrease in the costs associated with catheter-related adverse events. Rates and costs of catheter-related adverse events were based on estimates from the literature. Sensitivity analyses were performed by variation of the estimated mean LOS decrease in the SD and by variation of the estimates for incidence and costs related to catheter complications. Costs were measured in year 2000 US dollars, and differences were not assessed for statistical significance. RESULTS: Of 177 patients treated with i.v. vancomycin, 103 (58%) were eligible for conversion to p.o. linezolid and 55 (31%) were eligible for early discharge from the hospital with continuation of p.o. therapy. Early discharge was associated with a mean (SD) LOS decrease of 3.3 (2.9) days. Annual mean total cost savings in patients eligible for conversion from i.v. vancomycin to p.o. linezolid with early discharge were $294,750 (range, $35,730-$553,790). For cases eligible for inpatient conversion from i.v. vancomycin to p.o. linezolid therapy (n=48), the mean total annual cost difference was an increase of $6340 for p.o. linezolid (range, -$12,910 to $11,900). CONCLUSION: These results--although partly based on estimates from the literature, rather than direct measurements--support the use of p.o. linezolid with or without early discharge as a potential cost-savings alternative for eligible patients treated with a full course of i.v. vancomycin for suspected or confirmed MRSS infection. PMID- 14749154 TI - An observational, retrospective, cohort study of dosing patterns for rofecoxib and celecoxib in the treatment of arthritis. AB - OBJECTIVE: This study assessed prescribing patterns for rofecoxib and celecoxib in the treatment of osteoarthritis (OA) and rheumatoid arthritis (RA), as well as differences in prescribing patterns across physician specialties. METHODS: This was an observational, retrospective, cohort study of a large, US pharmacy claims database. Eligible patients were initiating therapy with rofecoxib or celecoxib and had succeeds, equals 90 days' supply of medication, as well as > or =1 medical claim specific to OA or RA between June 1, 2000, and May 31, 2001. Analyses were stratified according to diagnosis, prescribing physician specialty, and patient demographics. The main outcome measure was mean daily usage (ie, mean daily dose [milligrams]; mean number of pills per day; and mean daily consumption [denoted as DACON], calculated as daily dose divided by most frequently prescribed strength). This was primarily a descriptive study. Tests of statistical significance were not performed because the large sample size would have rendered small differences significant. RESULTS: A total of 58,574 patients with OA (81.8% [n=47,935]) or RA (18.2% [n=10,639]) received 220,627 prescriptions for rofecoxib or celecoxib (47.7% [n=27, 924] and 52.3% [n=30, 650] of patients, respectively) during the study period. Overall, the most frequently prescribed strengths were rofecoxib 25 mg and celecoxib 200 mg. In both OA and RA, the most frequently prescribed mean daily dose of rofecoxib was 25 mg. In OA, the most frequently prescribed mean daily dose of celecoxib was 200 mg; in RA, it was 400 mg. Both pills per day and DACON were higher for celecoxib than rofecoxib. The DACON for rofecoxib was unrelated to physician specialty. Rheumatologists prescribed celecoxib at 20% to 40% higher mean daily doses than did primary care physicians, orthopedic specialists, or other specialists. Regardless of physician specialty, the DACON appeared higher for patients with RA than OA, for men than women, and for younger (aged <65 years) than older patients. CONCLUSIONS: In this analysis, relative to the most frequently prescribed strength, celecoxib-treated patients with OA and RA had higher DACONs than rofecoxib-treated OA and RA patients across all subgroups. These observations may have economic implications in terms of direct effects on cost and the need for formularies to consider overall use patterns in addition to pill costs. However, these conclusions are limited by lack of clinical information (other than an OA or RA diagnosis), inability to ascertain actual use, and potential for selection bias. PMID- 14749157 TI - Effect of exposure to secondhand smoke on markers of inflammation: the ATTICA study. AB - PURPOSE: We sought to investigate the effect of secondhand smoke exposure on inflammatory markers related to cardiovascular disease. METHODS: During 2001 to 2002, we randomly selected a stratified (age-sex) sample of adults without clinical evidence of cardiovascular disease. Exposure to secondhand smoke (>30 minutes per day and > or =1 day per week) was recorded. Multivariate regression analysis was used to evaluate the effects of exposure to secondhand smoke on levels of C-reactive protein, fibrinogen, homocysteine, and oxidized low-density lipoprotein (LDL) cholesterol, and on white blood cell count. RESULTS: One hundred and thirty-seven (38%) of the 357 men who had never smoked and 211 (33%) of the 638 never-smoking women reported current exposure to secondhand smoke. Compared with those who were not exposed to secondhand smoke, those exposed more than 3 days per week had higher white blood cell counts (by 600 cells per microL; P = 0.02), as well as higher levels of C-reactive protein (by 0.08 mg/dL; P = 0.03), homocysteine (by 0.4 micromol/L; P = 0.002), fibrinogen (by 5.2 mg/dL; P = 0.4), and oxidized LDL cholesterol (by 3.3 mg/dL; P = 0.03), after adjusting for several potential confounders. CONCLUSION: Our findings suggest another pathophysiological mechanism by which exposure to secondhand smoke is associated with the development of atherosclerosis. PMID- 14749158 TI - Effects of sustained-release bupropion among persons interested in reducing but not quitting smoking. AB - PURPOSE: To determine whether sustained-release bupropion promotes smoking reduction leading to smoking cessation among persons who wish to reduce their amount of smoking, but who are unwilling to quit or who perceive themselves as being unable to quit. METHODS: Current smokers were assigned randomly to receive either sustained-release bupropion (150 mg twice daily) or matching placebo. During an initial 6-month smoking reduction phase, those who were willing to quit entered a 7-week cessation phase, during which study medication was continued. RESULTS: Four-week continuous abstinence rates were 14% (41/295) in the bupropion group and 8% (25/299) in the placebo group (P = 0.02) during treatment. However, this benefit did not continue after treatment was stopped; subsequent continuous abstinence rates were 7% (20/295) in the bupropion group and 5% (16/299) in the placebo group (P = 0.50). Similar proportions of subjects entered the cessation phase in both treatment groups (38% [n = 113] of those in the bupropion group and 34% [n = 101] of those in the placebo group), although the time until a cessation attempt was shorter for those taking bupropion (median, 64 days vs. 118 days, P = 0.008). The extent of smoking reduction (measured by urinary cotinine concentrations) among the 327 subjects who did not enter the cessation phase was significantly greater (P <0.05) in those treated with bupropion during the reduction treatment phase, but not during the month 12 follow-up visit (P = 0.25). CONCLUSION: Sustained-release bupropion, when used in smokers initially not willing to make a cessation attempt, can help sustain smoking reduction while subjects are on active medication, reduce the time until the next cessation attempt, and increase short-term abstinence rates. However, these benefits were modest and not sustained after bupropion was discontinued. PMID- 14749159 TI - Effect of folic acid fortification of food on homocysteine-related mortality. AB - PURPOSE: In 1998, the Food and Drug Administration mandated the fortification of food products with folic acid. The effect of this rule on mortality associated with homocysteine levels in patients with coronary artery disease is unknown. METHODS: We studied 2481 consecutive patients with coronary artery disease who underwent coronary angiography between 1994 and 1999, and who had baseline homocysteine measurements and at least 2 years of follow-up. Patients were divided into prefortification (1994 to 1997, n = 1595) and postfortification (1998 to 1999, n = 886) groups, as well as classified based on baseline homocysteine levels (normal to low, intermediate, and high). Homocysteine levels were measured by fluorescence polarization immunoassay. Mortality was determined by telephone survey or from a national Social Security database or hospital records. RESULTS: After implementation of the fortification rule, median homocysteine levels declined from 13.8 to 12.3 micromol/L (P <0.001), and the proportion of patients with high homocysteine levels (>15 micromol/L) decreased from 41% (n = 650) to 28% (n = 249) (P <0.001). Overall, homocysteine was a modest risk factor for mortality (adjusted relative risk [RR] = 1.03 per micromol/L; 95% confidence interval [CI]: 1.01 to 1.05; P = 0.006). There was no significant interaction between fortification status and homocysteine category with mortality (P for interaction = 0.85). Two-year mortality was reduced minimally (7.8% [n = 124] to 7.2% [n = 64]; RR = 0.93; 95% CI: 0.68 to 1.27; P = 0.63; adjusted RR = 0.97; 95% CI: 0.68 to 1.40), but was consistent with the expectation of a modest reduction in homocysteine levels. CONCLUSION: Homocysteine is an independent, graded risk factor for mortality. Homocysteine levels decreased modestly after the fortification of food with folic acid, but the effects on mortality were minor and likely attributable to other factors, indicating the need for more aggressive measures to reduce homocysteine associated cardiovascular risk. PMID- 14749160 TI - Antihistone and anti-double-stranded deoxyribonucleic acid antibodies are associated with renal disease in systemic lupus erythematosus. AB - PURPOSE: We sought to assess the nephritogenic antibody profile of patients with systemic lupus erythematosus (SLE), and to determine which antibodies were most useful in identifying patients at risk of nephritis. METHODS: We studied 199 patients with SLE, 78 of whom had lupus nephritis. We assayed serum samples for antibodies against chromatin components (double-stranded deoxyribonucleic acid [dsDNA], nucleosome, and histone), C1q, basement membrane components (laminin, fibronectin, and type IV collagen), ribonucleoprotein, and phospholipids. Correlations of these antibodies with disease activity (SLE Disease Activity Index) and nephropathy were assessed. Patients with no initial evidence of nephropathy were followed prospectively for 6 years. RESULTS: Antibodies against dsDNA, nucleosomes, histone, C1q, and basement membrane components were associated with disease activity (P <0.05). In a multivariate analysis, anti dsDNA antibodies (odds ratio [OR] = 6; 95% confidence interval [CI]: 2 to 24) and antihistone antibodies (OR = 9.4; 95% CI: 4 to 26) were associated with the presence of proliferative glomerulonephritis. In the prospective study, 7 (6%) of the 121 patients developed proliferative lupus glomerulonephritis after a mean of 6 years of follow-up. Patients with initial antihistone (26% [5/19] vs. 2% [2/95], P = 0.0004) and anti-dsDNA reactivity (6% [2/33] vs. 0% [0/67], P = 0.048) had a greater risk of developing proliferative glomerulonephritis than patients without these autoantibodies. CONCLUSION: In addition to routine anti dsDNA antibody assay, antihistone antibody measurement may be useful for identifying patients at increased risk of proliferative glomerulonephritis. PMID- 14749161 TI - Association of older age and female sex with inadequate reach of screening flexible sigmoidoscopy. AB - PURPOSE: Estimates of the sensitivity of screening sigmoidoscopy assume an adequate depth of insertion is reached. However, in clinical practice, the frequency that sigmoidoscopy reaches various lengths of the colon is not known. We assessed the frequency of inadequate reach (depth of <50 cm of the colon) in a large U.S. cohort, according to age and sex. METHODS: We performed a cross sectional study of 15,406 asymptomatic persons aged 50 years or older who underwent screening flexible sigmoidoscopy between April 1997 and October 2001 at sites participating in the Clinical Outcomes Research Initiative, which examines outcomes of endoscopy in "real life" settings. The maximum depth of insertion of the sigmoidoscope was measured in centimeters from the anus and classified as adequate (> or =50 cm) or inadequate (< 50 cm). Patient characteristics as well as procedure-related variables were also recorded. RESULTS: Eighteen percent (n = 2801) of subjects had an inadequate examination. In men, the percentage of inadequate examinations increased progressively with age, from 10% (343/3338) in those aged 50 to 59 years to 22% (53/248) in those aged 80 years or older (P <0.001). Inadequate examinations were more common in women at all ages, ranging from 19% (733/3798) in those aged 50 to 59 years to 32% (86/267) in those aged 80 years or older (P <0.001). These associations were confirmed in a multivariable analysis. CONCLUSION: Our finding that advancing age and female sex were independently associated with the risk of inadequate reach of screening sigmoidoscopy suggests that the sensitivity of sigmoidoscopy may be lower in these populations. Estimates of the benefits of sigmoidoscopy may need to be tailored to the age and sex of the patient. PMID- 14749162 TI - The end of the disease era. AB - The time has come to abandon disease as the focus of medical care. The changed spectrum of health, the complex interplay of biological and nonbiological factors, the aging population, and the interindividual variability in health priorities render medical care that is centered on the diagnosis and treatment of individual diseases at best out of date and at worst harmful. A primary focus on disease may inadvertently lead to undertreatment, overtreatment, or mistreatment. The numerous strategies that have evolved to address the limitations of the disease model, although laudable, are offered only to a select subset of persons and often further fragment care. Clinical decision making for all patients should be predicated on the attainment of individual goals and the identification and treatment of all modifiable biological and nonbiological factors, rather than solely on the diagnosis, treatment, or prevention of individual diseases. Anticipated arguments against a more integrated and individualized approach range from concerns about medicalization of life problems to "this is nothing new" and "resources would be better spent determining the underlying biological mechanisms." The perception that the disease model is "truth" rather than a previously useful model will be a barrier as well. Notwithstanding these barriers, medical care must evolve to meet the health care needs of patients in the 21st century. PMID- 14749163 TI - The demise of disease? I don't think so. PMID- 14749164 TI - Leapfrog and critical care: evidence- and reality-based intensive care for the 21st century. AB - In 2000, the Business Roundtable published its Leapfrog report, which contained suggestions for improving administration of critical care. The Leapfrog Group intends to influence health care policy by pressuring insurers and hospitals to implement its guidelines, and both internists and intensivists are likely to be affected if these recommendations are realized. This article outlines the Leapfrog standards for critical care and examines critically the evidence used to justify them. Aside from the guideline that all critically ill patients should be cared for by intensivists, Leapfrog's standards for critical care are based either on weak or no scientific evidence. Rather, most of the guidelines are grounded in common sense and rational extrapolation of the data; as such, they are a reasonable starting point for debate by physicians and policymakers about optimal methods of achieving intensivist-guided care of critically ill patients. PMID- 14749165 TI - Immunosuppressive therapy for idiopathic retroperitoneal fibrosis: a retrospective analysis of 26 cases. PMID- 14749166 TI - Cases from the Osler Medical Service at Johns Hopkins University. AB - A 47-year-old white woman with a history of stage III squamous cell carcinoma of the anus was transferred to Johns Hopkins Hospital for further evaluation of renal failure, hemolytic anemia, and thrombocytopenia. The patient was first diagnosed with squamous cell carcinoma of the anus 1 year before admission. She was treated with external beam radiation of the pelvis and two cycles of mitomycin C-based chemotherapy (a cumulative dose, 34 mg/m(2)). Her clinical course was complicated by Clostridium difficile colitis and myositis successfully treated with prednisone. Three months before admission, the patient developed dysuria. Her creatinine increased from normal to 1.7 mg/dL, and microscopic hematuria was present. A renal ultrasound and an abdominal computed tomographic scan showed no abnormalities or obstruction. One month before admission, she underwent a cystoscopy, which showed only radiation-induced changes in the bladder. Two weeks before admission, the patient became delirious and was taken to a hospital, where she was found to be anemic, with a hematocrit level of 23.7%, and thrombocytopenic with a platelet count of 110,000/mm(3). Her creatinine level was 5.9 mg/dL. Previous values of hematocrit, platelet count, and serum creatinine were normal. On admission at Johns Hopkins Hospital the patient had no complaints. She was afebrile on physical examination and had normal vital signs. Head, neck, chest, cardiovascular, and abdominal examinations were normal. There was skin pallor, but no echymoses or petechiae. She was alert and oriented with normal mental status. Her neurologic examination was normal. Laboratory data showed a white blood cell count of 6390/mm(3), a hematocrit level of 26.5%, and a platelet count of 26,000/mm(3). Her blood urea nitrogen level was 57 mg/dL, creatinine level was 4.0 mg/dL, and lactate dehydrogenase was 550 U/L (reference, 115 to 275 U/L). Urinalysis showed innumerable red blood cells and large protein. A peripheral blood smear showed fragmented red blood cells, schistocytes, no abnormal white blood cells, and few platelets. There was no radiographic or clinical evidence of relapse of her squamous cell carcinoma. What is the diagnosis? PMID- 14749167 TI - Secondhand smoke: the evidence of danger keeps growing. PMID- 14749168 TI - Smoking less as a treatment goal for those who cannot stop smoking. PMID- 14749169 TI - Look before you leap: how do intensivists improve care for critically ill patients? PMID- 14749170 TI - Aspergillus rib and vertebral osteomyelitis in a former intravenous drug user. PMID- 14749171 TI - Helicobacter pylori infection and idiopathic epilepsy. PMID- 14749172 TI - Hyperammonemia: elevated ammonia levels in multiple myeloma. PMID- 14749173 TI - Tremor, aphasia, and stuttering associated with Helicobacter pylori infection. PMID- 14749174 TI - By the numbers: data on medical school-based departments of internal medicine. PMID- 14749175 TI - Phylogenetic analysis of bovine viral diarrhea viruses using five different genetic regions. AB - Phylogenetic analysis of the five different regions (5' non-coding region (5'NCR), N(pro), E2, NS3 and NS5B-3'NCR) of 48 Japanese and reported bovine viral diarrhea virus (BVDV) genomes was performed. Japanese BVDVs were segregated into BVDV1 subdivided into six subgroups and BVDV2. One isolate, So CP/75, isolated in 1975 and previously proposed as subgroup 1e according to its 5'NCR sequence, was quite unique and formed an independent lineage in the tree of any region. Another isolate, 190CP, obtained from an experimental mucosal disease case was classified as subgroup 1e, defined by Becher et al. in the 5'NCR, N(pro) and E2 regions, whereas it was classified as subgroup 1a in the NS5B-3'NCR region. The genomic sequences of the American isolates ILLC and ILLNC obtained from the GenBank database were assigned into subgroup 1b in the 5'NCR, N(pro), E2 and NS5B-3'NCR regions, whereas they were assigned into subgroup 1a in the NS3 region, suggesting that recombination between the virus strains classified into different subgroups had occurred in an animal. These findings suggest that phylogenetic analysis of several genetic regions is useful for the further characterization of field BVDV isolates. PMID- 14749176 TI - Structure of a knockout mutant of influenza virus M1 protein that has altered activities in membrane binding, oligomerisation and binding to NEP (NS2). AB - The influenza virus M1 (matrix) protein forms the connection between the membrane component of the virus and its replication component eight ribonucleoprotein particles (RNPs). For this activity, M1 self-polymerises in the infected cell in order to pull glycoprotein containing membrane segments together. Later in the process of infection, M1 enters the nucleus and is active in the nuclear export process of newly made RNPs for virus particle assembly. The N-terminal domain (residues 1-164) of M1 carries the nuclear localisation sequence (NLS) motif and is important for membrane binding, self-polymerisation and nuclear export of RNPs. An NLS-mutant M1 has been used in functional studies in order to implicate the positive charges in the NLS in these three activities. In this paper, the crystal structure of the N-terminal domain of this NLS-mutant is determined and is found to be the same as that of the wild-type protein, clearly indicating that it is the absence of the positively charged residues of the NLS that causes the knock-out phenotype rather than a change in the overall structure of the mutant protein. PMID- 14749177 TI - Genetic characterization of a mumps virus isolate during passaging in the amniotic cavity of embryonated chicken eggs. AB - The aim of this study was the molecular characterization of a historical mumps isolate (an alleged individual sample). After RNA extraction and cDNA synthesis, selective nested PCR amplification with specific primers, automated DNA sequencing and RFLP analyses were performed. The relative ratios of the detected virus sequences were determined by GeneScan electrophoresis. Phylogenetic tree based on the 316 nucleotide region of the SH gene of the mumps virus was generated by the neighbor-joining method. Results obtained by the described molecular approach show: (a) there are two mumps virus variants, A and B, detected in the fourth passage of wild type virus in the amniotic cavity of embryonated chicken eggs (ECE); (b) variants A and B belong to different genotypes; (c) variants A and B differ in the HN and NP genes which code for amino acid sequences comprising immunogenic epitopes; (d) variant B contains one or more minor variants. We discuss whether the observed differences between the two variants are a consequence of natural heterogeneity or of laboratory contamination in the early passages. PMID- 14749178 TI - Development of a reconstitution system for Rinderpest virus RNA synthesis in vitro. AB - The RNA dependent RNA polymerase of Rinderpest virus consists of two subunits-the large protein (L) and the phosphoprotein (P), where L is thought to be responsible for the catalytic activities in association with P protein which plays multiple roles in transcription and replication. The nucleocapsid protein (N) is necessary for encapsidation of genomic RNA, which is required as N-P complex. To understand the different steps of transcription and replication as well as the roles played by the three proteins, an in vitro reconstitution system for RNA synthesis is necessary which is not available for any morbillivirus. We describe here, an in vitro reconstitution system for transcription and replication of Rinderpest virus utilizing a synthetic, positive sense N-RNA minigenome template, free of endogenous viral polymerase proteins and recombinant viral proteins (P+L and P+N) expressed in insect cells by recombinant baculoviruses. We show that although L-P complex is sufficient to synthesize negative sense minigenome RNA, soluble N protein is necessary for encapsidation of RNA as well as synthesis of (+) sense leader RNA and (+) sense minigenome RNA. PMID- 14749179 TI - Effect of single amino acid mutations in the conserved GDNQ motif of L protein of Rinderpest virus on RNA synthesis in vitro and in vivo. AB - The paramyxovirus RNA-dependent RNA polymerase consists of two subunits, the transcription co-factor phosphoprotein P and the large protein L, which possesses all the catalytic functions such as RNA synthesis (both transcription replication), methylation, capping and polyadenylation. The L protein has high sequence homology among the negative sense RNA viruses. The domains and residues on the L protein involved in the above-mentioned activities are not well defined, although the role of conserved GDNQ motif of the putative catalytic centre of L protein of few related viruses have been examined. In order to gain insight into the role played by the GDNQ motif of the L protein of Rinderpest virus (RPV), we have examined mutations at each amino acid in this motif of the L protein of Rinderpest virus and tested the biological activity in vivo and in vitro. Site directed mutants were generated and transiently expressed in mammalian cells and were shown to interact with P protein similar to wild type L. The biological activity of mutant L proteins has been tested in an in vitro reconstituted system capable of carrying out cell-free RNA synthesis on synthetic Rinderpest N-RNA template. Further, the role played by individual amino acids has also been defined in vivo using an in vivo minigenome replication/transcription system which indicated the importance of this conserved sequence in viral RNA synthesis. PMID- 14749180 TI - Isolation and identification of an enterovirus 77 recovered from a refugee child from Kosovo, and characterization of the complete virus genome. AB - The complete nucleotide sequence of an enterovirus 77 isolate is reported. The virus designated FR/CF496-99 (France/Clermont-Ferrand 496-1999) was recovered from the feces of a 4-year-old child hospitalized for Salmonella gastroenteritis. The virus was identified by a molecular typing assay based on the genomic sequence encoding the VP1 capsid protein. The phylogenetic analysis based on the VP1 sequence demonstrated that the enterovirus isolated in the child clustered with viruses included in the human enterovirus B species (HEV-B) and was most closely related to enterovirus 77. A sliding window analysis of the complete genome showed an overall nucleotide similarity >80% between the P3 genomic region of the FR/CF496-99 isolate and that of the echovirus 30 prototype strain. A comparative analysis based on partial 3D(pol) sequences showed that the FR/CF496 99 virus was more closely related to recent enteroviruses from different serotypes and different geographical areas than to the prototype strains collected in the 1950s. This suggests that, in this enterovirus, the 3D(pol) encoding sequence is of recent origin. PMID- 14749181 TI - The C-terminal domain of measles virus nucleoprotein belongs to the class of intrinsically disordered proteins that fold upon binding to their physiological partner. AB - The nucleoprotein of measles virus consists of an N-terminal domain, N(CORE) (aa 1-400), resistant to proteolysis, and a C-terminal domain, N(TAIL) (aa 401-525), hypersensitive to proteolysis and not visible by electron microscopy. Using two complementary computational approaches, we predict that N(TAIL) belongs to the class of natively unfolded proteins. Using different biochemical and biophysical approaches, we show that N(TAIL) is indeed unstructured in solution. In particular, the spectroscopic and hydrodynamic properties of N(TAIL) indicate that this protein domain belongs to the premolten globule subfamily within the class of intrinsically disordered proteins. The isolated N(TAIL) domain was shown to be able to bind to its physiological partner, the phosphoprotein (P), and to undergo an induced folding upon binding to the C-terminal moiety of P [J. Biol. Chem. 278 (2003) 18638]. Using a computational analysis, we have identified within N(TAIL) a putative alpha-helical molecular recognition element (alpha MoRE, aa 488-499), which could be involved in binding to P via induced folding. We report the bacterial expression and purification of a truncated form of N(TAIL) (N(TAIL2), aa 401-488) devoid of the alpha-MoRE. We show that N(TAIL2) has lost the ability to bind to P, thus supporting the hypothesis that the alpha MoRE may play a role in binding to P. We have further analyzed the alpha-helical propensities of N(TAIL2) and N(TAIL) using circular dichroism in the presence of 2,2,2-trifluoroethanol. We show that N(TAIL2) has a lower alpha-helical potential compared to N(TAIL), thus suggesting that the alpha-MoRE may be indeed involved in the induced folding of N(TAIL). PMID- 14749182 TI - Transient disulfide bonds formation in conformational maturation of influenza virus nucleocapsid protein (NP). AB - It has been previously shown that influenza virus nucleocapsid protein (NP) forms homooligomers in vivo. Our analyses revealed that the reducing agent dithiothreitol (DTT) introduced in pulse labeling period prevented further formation of native NP-oligomers. The shortly pulse-labeled non-reduced newly synthesized NP possessed a relatively faster mobility in non-reducing PAGE and a higher resistance to protease than the reduced one. These data suggest that there is an early disulfide-dependent step in NP maturation and that the newly synthesized NP possesses the intrachain disulfide bonds. In contrast to the newly synthesized NP, the non-reduced chased NP possessed the same mobility in non reducing PAGE and the same sensitivity to protease as the reduced NP. DTT introduced in the chase period did not prevent NP-oligomers formation and did not destabilize already formed NP-oligomers. This suggests that the chased NP monomers and NP-oligomers do not contain intrachain nor interchain disulfide bonds. It was also shown that the non-reduced newly synthesized NP could not form NP-NP complexes in vitro, and acquired such ability only after reducing. The possibility is discussed that there are several stages in the maturation of NP: the initial formation of intrachain disulfide-linked NP and conversion into disulfide-free NP, which forms non-covalently stabilized NP-oligomers. Early intrachain disulfide bonds may be necessary for the prevention of early spontaneous NP-NP association. PMID- 14749183 TI - An oligosaccharide at the C-terminus of the F-specific domain in the stalk of the human parainfluenza virus 3 hemagglutinin-neuraminidase modulates fusion. AB - The promotion of membrane fusion by the fusion (F) protein of human parainfluenza virus 3 (hPIV3) is dependent on a virus-specific contribution from the hemagglutinin-neuraminidase (HN) protein. By evaluation of chimeric hPIV3 Newcastle disease virus (NDV) HN proteins, we have previously shown that hPIV3-F specificity is determined by a domain that extends from the middle of the membrane anchor to the 82nd residue in the ectodomain [Virology 209, (1995) 457; Arch. Virol. 13 (1997) 115]. If the corresponding NDV-derived residues replace the two C-terminal residues in this domain, no fusion is detected. However, these substitutions restore a glycosylation site present in NDV HN, but not in hPIV3 HN. Deletion of this site from a nested set of chimeras with hPIV3-derived N terminal portions of decreasing length partially restores fusion, suggesting that an oligosaccharide near the top of hPIV3 HN stalk modulates fusion. In addition, further mutational analyses show that a chimera with only 125 N-terminal hPIV3 derived residues (72 in the stalk) actually promotes fusion more efficiently than the wt protein. These findings localize the C-terminus of the F-specific domain in hPIV3 HN a full 10 residues closer to the membrane than previously shown. PMID- 14749184 TI - Phylogenetic relationships in the Lymphocryptovirus genus of the Gammaherpesvirinae. AB - Complete DNA sequences were determined for the glycoprotein B (gB) genes of four viruses from the genus Lymphocryptovirus, whose hosts had been assigned as baboon, orangutan, chimpanzee and gorilla. Together with published sequences for the gB genes of three lymphocryptoviruses, namely the human pathogen Epstein-Barr virus (EBV), a rhesus monkey virus and a marmoset virus, the sequences were used to investigate evolutionary relationships in the genus. The chimpanzee and orangutan viruses' sequences were found to be so close that it is unlikely both represent natural infections in these hosts. Phylogenetic analyses showed that the New World marmoset virus lineage formed a sister clade to that of the Old World viruses, consistent with a cospeciational separation. Within the Old World virus group, resolution of branching pattern was incomplete, and suggestive of a complex history. In particular, it was inferred that separation of the EBV lineage from that of the gorilla virus plus the chimpanzee/orangutan virus may have predated separation of the present day host species. PMID- 14749185 TI - Efficient propagation of single gene deleted recombinant Sendai virus vectors. AB - Recombinant Sendai virus vectors (SeVV) have become an attractive tool for basic virological as well as for gene transfer studies. However, to (i) reduce the cellular injury induced by basic recombinant SeV vectors (encoding all six SeV genes as being present in SeV wild-type (wt) genomes) and to (ii) improve SeV vector safety, deletions of viral genes are necessary for the construction of superior SeVV generations. As a strong expression system recombinant replication incompetent adenoviruses, coding for SeV proteins hemagglutinin-neuraminidase (HN), fusion (F), or matrix (M), were generated and successfully employed for the propagation of single gene deleted (DeltaHN, DeltaF, DeltaM) recombinant SeVV. Further investigations of the propagation procedures required for single gene deleted recombinant SeVV demonstrated (i) modifications of the cell culture medium composition as well as (ii) incubation with vitamin E as crucial steps for the enhancement of SeVV-DeltaHN, -DeltaF, or -DeltaM viral particle yield. Such optimized propagation procedures even led to a successful propagation of HN deleted viral particles (SeVV-DeltaHN), which has not been reported before. PMID- 14749186 TI - The human parainfluenza virus type 3 (HPIV 3) C protein inhibits viral transcription. AB - The C protein of human parainfluenza virus type 3 (HPIV 3), like other paramyxovirus C proteins, is synthesized from an alternate open reading frame (ORF) encoded within the phosphoprotein (P) mRNA, in addition, to two other proteins, namely D and V, which arise from the same mRNA by a process of transcriptional editing. The precise role of the C, D, and V proteins in viral transcription and replication, and their interaction, if any, with other viral proteins remains unknown. To ascertain the role of the C protein, we have examined its effect on transcription using an HPIV 3 minigenome construct and monitoring the luciferase reporter gene expression. Our results demonstrate that the HPIV 3 C protein effectively inhibits minigenome transcription in a dose dependent manner. Interestingly, the Sendai virus (Se-V) C protein was also capable of inducing an inhibitory effect on the HPIV 3 minigenome transcription, thus demonstrating a heterologous interaction. A coiled-coil motif within the C protein has been identified, and a deletion mutant within this motif abrogated the inhibitory effect significantly thereby implying that oligomerization of the C protein may be involved in inhibition of transcription. PMID- 14749187 TI - Influenza H5 virus escape mutants: immune protection and antibody production in mice. AB - Avian H5N1 influenza A viruses are considered to be of high pandemic potential as they are able to cross the avian-human species barrier and cause disease in humans. In the present study we assessed the impact of amino acid substitutions in the hemagglutinin (HA) of antigenic escape mutants of influenza A/Mallard/Pennsylvania/10218/84 (H5N2) (Mld/PA/84-MA) virus on the level of neutralizing antibodies and the ability to protect mice against challenge with the wild type H5 influenza virus. beta-Propiolactone-inactivated vaccines prepared from eight different H5 escape mutants could be separated into two groups based on levels of protection. One group of escape mutants [m46(7), m46(7) 24B9, m46(7)-55, and m46(7)-55-24B9] was characterized by providing high levels of protection (90.0-95.4% survival) to mice against subsequent challenge with 5 LD(50) of wild type Mld/PA/84-MA virus. The other group of escape mutants [m176/26, m55(2), m55(2)-24B9, and m24B9-176/26] provided moderate level of protection (57.1-66.6% survival) in mice. Analysis of the amino acid substitutions in the HA revealed that two amino acid changes in antigenic site B of the HA molecule (D(126)-->N and K(152)-->N) were associated for decreases in the levels of antibody and the immune protection afforded by vaccination with these H5 virus escape mutants. The phenotypic effects of mutations in HA gene of H5 virus may be of importance to appraise the extent and direction of H5 influenza viruses antigenic evolution. PMID- 14749188 TI - Understanding pain sufferers: the role of cognitive processes. PMID- 14749191 TI - A randomized, placebo-controlled trial of intradiscal electrothermal therapy for the treatment of discogenic low back pain. AB - BACKGROUND: Intradiscal electrothermal therapy (IDET) is a treatment for discogenic low back pain the efficacy of which has not been rigorously tested. PURPOSE: To compare the efficacy of IDET with that of a placebo treatment. STUDY DESIGN/SETTING: Randomized, placebo-controlled, prospective trial. PATIENT SAMPLE: Patients were recruited by referral and the media. No inducements were provided to any patient in order to have them participate. Of 1,360 individuals who were prepared to submit to randomization, 260 were found potentially eligible after clinical examination and 64 became eligible after discography. All had discogenic low back pain lasting longer than 6 months, with no comorbidity. Thirty-seven were allocated to IDET and 27 to sham treatment. Both groups were satisfactorily matched for demographic and clinical features. METHODS: IDET was performed using a standard protocol, in which the posterior annulus of the painful disc was heated to 90 C. Sham therapy consisted of introducing a needle onto the disc and exposing the patient to the same visual and auditory environment as for a real procedure. Thirty-two (85%) of the patients randomized to the IDET group and 24 (89%) of those assigned to the sham group complied fully with the protocol of the study, and complete follow-up data are available for all of these patients. OUTCOME MEASURES: The principal outcome measures were pain and disability, assessed using a visual analog scale for pain, the Short Form (SF) 36, and the Oswestry disability scale. RESULTS: Patients in both groups exhibited improvements, but mean improvements in pain, disability and depression were significantly greater in the group treated with IDET. More patients deteriorated when subjected to sham treatment, whereas a greater proportion showed improvements in pain when treated with IDET. The number needed to treat, to achieve 75% relief of pain, was five. Whereas approximately 40% of the patients achieved greater than 50% relief of their pain, approximately 50% of the patients experienced no appreciable benefit. CONCLUSIONS: Nonspecific factors associated with the procedure account for a proportion of the apparent efficacy of IDET, but its efficacy cannot be attributed wholly to a placebo effect. The results of this trial cannot be generalized to patients who do not fit the strict inclusion criteria of this study, but IDET appears to provide worthwhile relief in a small proportion of strictly defined patients undergoing this treatment for intractable low back pain. PMID- 14749190 TI - Transplantation of oligodendrocyte precursors and sonic hedgehog results in improved function and white matter sparing in the spinal cords of adult rats after contusion. AB - BACKGROUND CONTEXT: A substantial cause of neurological disability in spinal cord injury is oligodendrocyte death leading to demyelination and axonal degeneration. Rescuing oligodendrocytes and preserving myelin is expected to result in significant improvement in functional outcome after spinal cord injury. Although previous investigators have used cellular transplantation of xenografted pluripotent embryonic stem cells and observed improved functional outcome, these transplants have required steroid administration and only a minority of these cells develop into oligodendrocytes. PURPOSE: The objective of the present study was to determine whether allografts of oligodendrocyte precursors transplanted into an area of incomplete spinal cord contusion would improve behavioral and electrophysiological measures of spinal cord function. Additional treatment incorporated the use of the glycoprotein molecule Sonic hedgehog (Shh), which has been shown to play a critical role in oligodendroglial development and induce proliferation of endogenous neural precursors after spinal cord injury. SETTING: Laboratory study. METHODS: Moderate spinal cord contusion injury was produced in 39 adult rats at T9-T10. Ten animals died during the course of the study. Nine rats served as contusion controls (Group 1). Six rats were treated with oligodendrocyte precursor transplantation 5 days after injury (Group 2). The transplanted cells were isolated from newborn rat pups using immunopanning techniques. Another eight rats received an injection of recombinant Shh along with the oligodendrocyte precursors (Group 3), while six more rats were treated with Shh alone (Group 4). Eight additional rats received only T9 laminectomies to serve as noninjured controls (Group 0). Animals were followed for 28 days. RESULTS: After an initial complete hindlimb paralysis, rats of all groups receiving a contusive injury recovered substantial function within 1 week. By 28 days, rats in Groups 2 and 3 scored 4.7 and 5.8 points better on the Basso, Beattie, Bresnahan (BBB) open field locomotor score than rats in group 1 (Groups 2 and 3=18.2 and 19.4 points, respectively, after 28 days vs. Group 1=13.6 points; p=.015). Rats in Group 4 scored no better than those in Group 1 (BBB=16.4). Motor evoked potential (MEP) recordings revealed a strong trend towards significant improvement in latency measurements in all treatment groups compared with controls at 28 days, although three animals in Group 1 and two animals in Group 3 were not recordable. Histological examination demonstrated significantly more spared white matter in the same groups that correlated with the improvements in BBB scores and MEP latencies. Immunohistochemical analysis showed the survival, proliferation and migration of the transplanted cells, as well as the induction of proliferating endogenous neural precursor cells in animals treated with Shh. CONCLUSIONS: These findings suggest that the transplantation of oligodendrocyte precursors may improve axonal conduction and spinal cord function in the injured spinal cord. The benefits seem more pronounced with the addition of Shh, and the addition of Shh alone results in the proliferation of an endogenous population of neural precursor cells. PMID- 14749192 TI - The effect of alendronate sodium on spinal fusion: a rabbit model. AB - BACKGROUND CONTEXT: Bisphosphonates affect bone remodeling and increase bone mass through the inhibition of osteoclasts. Their effect on osteoblasts, and the balance between osteoblastic and osteoclastic activity on bone turnover and healing, is not completely understood. Specifically, the effect of bisphosphonates on spinal fusion has yet to be determined. With the increasing use of bisphosphonates in the elderly population, this effect needs to be delineated. PURPOSE: To evaluate the effect of alendronate sodium after an intertransverse process spinal fusion in a rabbit model. STUDY DESIGN/SETTING: Randomized double-blinded in vivo study of the effect of alendronate sodium in a spinal fusion model. METHODS: Fifty New Zealand white rabbits underwent a posterolateral L5-L6 intertransverse process arthrodesis with autogenous iliac crest bone graft. The rabbits were then randomly divided into two groups. Group I received 3 cc of saline placebo per oral gavage, and Group II received 200 micrograms (approximately 0.05 mg/kg/day) of alendronate sodium dissolved in 3 cc of saline per day for 8 weeks. Upon completion, the rabbits were sacrificed and the lumbar spines harvested, radiographed and graded for motion across the fusion site with manual palpation. Two independent pathologists then prepared and sectioned each left and right fusion mass. Three random x10 fields were examined and graded for both the cephalad and caudad ends of each section (516 fields). Fusion quality was graded using an established histological scoring scale (score 0 to 7 based on fibrous and bone content of the fusion mass). RESULTS: Two rabbits died on the day of operation, and 48 rabbits survived the operation. Five additional rabbits died within the first 2 postoperative weeks. Thus, 43 rabbits (21 in Group I, 22 in Group II) completed the 8-week course of treatment. Grading each side separately, 26 of 42 fusion masses (62%) in Group I and 24 of 44 fusion masses (55%) in Group II had radiographic evidence of fusion (p=.76). With gross palpation, 11 of 21 motion segments (52%) in Group I versus 13 of 22 motion segments (59%) in Group II were determined to have a solid fusion (p=.76). Histologically, Group I had a higher median score (6.0; range, 0 to 7 vs. 1.0; range, 0 to 7; p<.0001) and a higher fusion rate (76% vs. 45%; p=.004) than Group II. CONCLUSIONS: Alendronate sodium appears to inhibit or delay bone fusion in a rabbit model. Presumably, this occurs as a result of uncoupling the balanced osteoclastic and osteoblastic activity inherent to bone healing. These findings suggest that a discontinuance of alendronate sodium postoperatively during the acute fusion period may be warranted. PMID- 14749193 TI - Cell-based tissue engineering for the intervertebral disc: in vitro studies of human disc cell gene expression and matrix production within selected cell carriers. AB - BACKGROUND CONTEXT: Little is known about how disc cells attach, proliferate and form extracellular matrix (ECM) within carrier materials. Such information is needed to help formulate criteria for successful cell-carrier interactions in tissue engineering. PURPOSE: To compare proliferation, ECM production and gene expression in annulus cells cultured in a variety of cell carrier materials with potential application in tissue engineering of the disc. STUDY DESIGN: Human intervertebral disc cells from the annulus were used in a prospective study of proliferation, ECM production and gene expression within selected cell carriers. METHODS: Annulus cells from discs of 29 individuals were tested in collagen sponge, collagen gel, agarose, alginate or fibrin gel formulations. In situ hybridization assessed ECM gene expression of Types I and II collagen, aggrecan and chondroitin-6 sulfotransferase. Cell proliferation, cell shape, attachment and ECM production were evaluated. RESULTS: Collagen sponges provided the best microenvironment for disc cell ECM production and gene expression. Although collagen gels often could support good cell growth, such constructs did not result in either abundant ECM production or ECM gene expression, as shown by in situ hybridization. Growth and ECM production and gene expression in alginate, agarose and fibrin microenvironments were inferior. CONCLUSIONS: Tissue engineering techniques open new therapeutic possibilities for use of autologous disc cells, but fundamental questions on how these cells interact with cell carriers are unexplored. Results provide novel data on disc cell gene expression within diverse microenvironments. The collagen sponge proved to be a superior microenvironment. PMID- 14749194 TI - National trends in nonoperative care for nonspecific back pain. AB - BACKGROUND CONTEXT: Few empirical data are available that document changes in population-based rates for the evaluation and treatment of nonspecific back pain. PURPOSE: To determine the extent of change in the pattern of outpatient evaluation and treatment of nonspecific low back pain in the United States between 1987 and 1997. STUDY DESIGN AND SETTING: The 1987 National Medical Expenditure Survey and the 1997 Medical Expenditure Panel Survey, two nationally representative surveys with similar sampling methods and questions, were used. PATIENT SAMPLE: Noninstitutionalized adults in the United States. OUTCOME MEASURES: Changes in rates of any health service for nonspecific back pain and occurrence of provider-specific care and types of services provided. Changes in the prescription of specific medication classes (ie, nonsteroidal anti inflammatory drugs [NSAIDs], muscle relaxants, nonnarcotic and narcotic analgesics) were also investigated. RESULTS: Overall rate for outpatient treatment for nonspecific back pain in the US population was relatively stable over the decade (4.48% in 1987, 4.53% in 1997, p=.85). Among those receiving care, the proportion receiving physician care increased from 64% in 1987 to 74% in 1997 (p<.001), whereas those obtaining care from physical therapists increased from 5% to 9% during the same time period (p<.01). The proportion of respondents receiving NSAIDs, muscle relaxants, nonnarcotic analgesics and narcotic analgesics remained stable. However, the mean number of patient visits in which these medications were prescribed increased from 2.0 to 3.9 over the decade (p<.001). The proportion of individuals receiving chiropractic care (p<.01) and X rays (p<.001) were lower in 1997 than 1987. CONCLUSIONS: The national pattern of health care for nonspecific low back pain observed in the present study serves as a basis for future investigations into the management of this major public health problem. Findings suggest that perhaps a duplication of care is partly responsible for the high degree of health care utilization in this population. PMID- 14749195 TI - Spine loading in patients with low back pain during asymmetric lifting exertions. AB - BACKGROUND CONTEXT: Recurrent low back pain (LBP) is a common and costly problem that might be related to increased spine loads in those with LBP. However, we know little about how the spine is loaded when those with LBP perform lifting exertions. PURPOSE: Document spine loading patterns of patients with LBP performing symmetric and asymmetric lifting exertions compared with asymptomatic individuals performing the same tasks. STUDY DESIGN: Spine loadings during lifting exertions that varied in asymmetric origin as well as horizontal and vertical distance from the spine were compared between asymptomatic subjects and patients with LBP. METHODS: Sixty-two patients with LBP and 61 asymptomatic individuals performed a variety of lifting exertions that varied in lift origin horizontal and vertical position (region), lift asymmetry position and weight lifted. An electromyography-assisted model was used to evaluate spine loading in each subject during the lifting exertions. Differences in spine loading between the LBP and asymptomatic subjects were noted as a function of the experimental variables. RESULTS: Patients with LBP experienced greater spine compression and shear forces when performing lifting tasks compared with asymptomatic individuals. The least taxing conditions resulted in some of the greatest differences between LBP and asymptomatic individuals. CONCLUSIONS: Greater levels of antagonistic muscle coactivation resulted in increases in spine loading for patients with LBP. Specific lifting conditions that tend to exacerbate loading can be identified by means of physical workplace requirements. These findings may impact acceptable return-to-work conditions for those with LBP. PMID- 14749196 TI - Distribution of anterior cortical shear strain after a thoracic wedge compression fracture. AB - BACKGROUND CONTEXT: Vertebral compression fractures (VCFs) are a common clinical problem and may follow trauma or be pathological. Osteoporosis increases susceptibility to fracture by reducing bone mass and weakening bone architecture. Approximately 2.5 million osteoporotic fractures occur worldwide annually, usually involving the vertebrae, wrist and hip. In the United States 700,000 VCFs occur annually, causing significant morbidity, mortality and economic burden. An initial VCF often leads to subsequent VCFs. The strain distribution along the anterior cortex, the major load-bearing pathway in flexion, may be predictive of impending VCF. Regions of high strain distribution are likely to experience secondary fracture. PURPOSE: To investigate the distribution of anterior cortical strain at, above and below an experimentally created index VCF to determine the vertebral body at risk of secondary fracture. STUDY DESIGN: In vitro experimental study using cadaveric thoracic spinal segments. METHODS: Seventeen thoracic spines underwent dual-energy X-ray absorptiometry (DEXA) to assess bone mineral density and were divided into T1-T3 (Subsegment 1), T4-T6 (Subsegment 2), T7-T9 (Subsegment 3) and T10-T12 (Subsegment 4). Rectangular rosette strain gauges were applied to the anterior cortices of the vertebrae of each subsegment (vertebrae in each specimen were denoted V1-superior, V2-intermediate and V3-inferior). V1 and V3 were partially embedded into polyester resin blocks, which were used to mount the specimens in a materials testing machine. Nondestructive predefect testing was performed in compression at 125 N and 250 N, followed by flexion at 1.25 Nm and 2.5 Nm. To ensure fracture reproducibility, V2 of each specimen had a trabecular defect created to a volume of 21.3+/-4.4% of the V2 centrum. Postdefect nondestructive compression and flexion were then performed in a manner similar to the predefect tests, followed by destructive testing in flexion. Anterior cortical shear strain on V1, V2 and V3, applied moments and applied flexion angle were all measured and analyzed. RESULTS: A VCF occurred in 55 of the 59 subsegments. Fifty-one VCF (93%) were seen in V2 and 4 VCF (7%) were seen in V1. After the creation of the trabecular defect, the shear strain on V2 increased, but a comparison of the postdefect with the predefect nondestructive tests showed no significant differences. The pre- and postdefect shear strain distribution in compression and flexion was V1strain>V3strain>V2strain. Shear strain at failure was highest on V2, and in all subsegments there were significant differences between V2 and V3 (p<.05). In all subsegments there were no significant differences between V2 and V1 (p>.05) at failure with the exception of Subsegment 1 where V2 and V1 were significantly different (p<.05). The predominant strain pattern at failure was (V2strain>V1strain>V3strain V2strain>>V3strain). Using shear strain as the codeterminant of peak moment with bending stiffness and applied angle at failure, the strain on V1 was the greatest predictor (p=.0084; R2=0.78). These findings suggest that the events leading to a secondary fracture probably start before the index VCF occurs and continue with loading beyond the index VCF. CONCLUSION: Anterior cortical strain is concentrated at the apex of a thoracic kyphotic curve. The vertebral body immediately above the index VCF has the next highest amount of strain and therefore the highest risk of secondary fracture. PMID- 14749197 TI - Percutaneous plasma decompression alters cytokine expression in injured porcine intervertebral discs. AB - BACKGROUND CONTEXT: Discectomy is a surgical technique commonly used to treat bulging or herniated discs causing nerve root compression. Clinical data suggest discectomy may also help patients with contained discs and no clear neural compromise. However, the mechanisms of clinical efficacy are uncertain, and consequently bases for treatment optimization are limited. PURPOSE: To determine the effect of percutaneous plasma decompression on the histologic, morphologic, biochemical and biomechanical features of degenerating intervertebral discs. STUDY DESIGN: An adult porcine model of disc degeneration was used to establish a degenerative baseline against which to evaluate discectomy efficacy. OUTCOME MEASURES: Cytokines interleukin (IL)-1, IL-6, IL-8, and tumor necrosis factor (TNF)-alpha were measured from tissue samples using enzyme-linked immunosorbent assay. Histology and morphology images were rated for degenerative findings (of cells and matrix) in both the nucleus and annulus. Proteoglycan content was determined, and intact specimen stiffness and flexibility were measured biomechanically. Magnetic resonance images were collected for biomechanical specimens. METHODS: Using a retroperitoneal surgical approach, stab incisions were made in four or five lumbar discs per spine in 12 minipigs. Animals were allocated into one of three groups: 6-week recovery, 12-week recovery and percutaneous plasma decompression using an electrosurgical device at 6 weeks with recovery for 6 additional weeks. Four additional animals served as controls. RESULTS: Discs treated with discectomy had a significant increase in IL-8 and a decrease in IL-1 as compared with the 12-week, nontreated discs. There were no significant differences in morphologic and biomechanical parameters or proteoglycan content between treated discs and time-matched, nontreated discs. CONCLUSIONS: Our results demonstrate that percutaneous plasma discectomy alters the expression of inflammatory cytokines in degenerated discs, leading to a decrease in IL-1 and an increase in IL-8. Whereas both IL-1 and IL-8 have hyperalgesic properties, IL-1 is likely to be a more important pathophysiologic factor in painful disc disorders than IL-8. Therefore, the alteration in cytokine expression that we observed is consistent with this effect as a mechanism of pain relief after discectomy. In addition, given that IL-1 is catabolic in injured tissue and IL-8 is anabolic, our results suggest that a percutaneous plasma discectomy may be capable of initiating a repair response in the disc. PMID- 14749198 TI - Bone ingrowth characteristics of porous tantalum and carbon fiber interbody devices: an experimental study in pigs. AB - BACKGROUND CONTEXT: In preclinical and clinical joint replacement applications, porous tantalum has been shown to be osteoconductive and effective for biological fixation. Relatively little research has been undertaken to investigate the porous tantalum implants for potential application in intervertebral spinal fusion. PURPOSE: The current study was designed to assess the radiographic and histological performance of porous tantalum and carbon fiber devices in the porcine anterior lumbar interbody fusion (ALIF) model. STUDY DESIGN: A total of 10 Danish Landrace pigs underwent a three-level anterior intervertebral lumbar arthrodeses at L2-L3, L4-L5 and L6-L7. Each level was randomly allocated to one of three implants: a solid piece of porous tantalum, a porous tantalum ring packed with autograft or a carbon fiber cage, likewise packed with autograft. Two staples for fixation were supplemented in front of implant. METHODS: Pigs were sacrificed 3 months after operation. Specimens were evaluated by plain radiography, conventional tomography and histology. RESULTS: Bone graft filled into the central hole of the porous tantalum ring was less than that of the carbon fiber cage (p<.001). Radiolucencies around the porous tantalum solid were significantly higher than the carbon fiber cage (p=.02) and were not different between the porous tantalum ring and the carbon fiber cage. The bone volume in the hole of implants, within the pores of the porous tantalum and in the implant interface did not differ between implants. Bone volume in the hole of the porous tantalum ring did not differ from that of the adjacent vertebral bone; however, it was significantly different in the carbon fiber cage and the adjacent vertebral bone (p=.005). CONCLUSIONS: In this porcine ALIF model, the radiographic and histological appearances of the porous tantalum ring were equivalent to those of the carbon fiber cage. The high presence of radiolucencies and fibrous tissue layer at the vertebrae-implant interface suggests that an initial stabilizing biomechanical environment is important in order to achieve bone ingrowth in the interbody fusion devices in this ALIF model. PMID- 14749199 TI - Exercise as a treatment for chronic low back pain. AB - BACKGROUND CONTEXT: Exercise is a widely prescribed treatment for chronic low back pain, with demonstrated effectiveness for improving function and work. PURPOSE: The goal of this article is to review several key aspects about the safety and efficacy of exercise that may help clinicians understand its utility in treating chronic back pain. STUDY DESIGN/SETTING: A computerized literature search of MEDLINE was conducted using "exercise," "fitness," "back pain," "backache" and "rehabilitation" as search words. Identified abstracts were scanned, and useful articles were acquired for further review. Additional references were acquired through the personal collections of research papers possessed by the authors and by reviewing prior review articles on this subject. These final papers were scrutinized for data relevant to the key aspects about exercise covered in this article. RESULTS: For people with acute, subacute or chronic low back pain, there is no evidence that exercise increases the risk of additional back problems or work disability. To the contrary, current medical literature suggests that exercise has either a neutral effect or may slightly reduce risk of future back injuries. Exercise can be prescribed for patients with chronic low back pain with three distinct goals. The first and most obvious goal is to improve or eliminate impairments in back flexibility and strength, and improve performance of endurance activities. There is a large body of evidence confirming that this goal can be accomplished for a majority of patients with chronic low back pain. The second goal of exercise is to reduce the intensity of back pain. Most studies of exercise have noted overall reduction in back pain intensity that ranges from 10% to 50% after exercise treatment. The third goal of exercise is to reduce back pain-related disability through a process of desensitization of fears and concerns, altering pain attitudes and beliefs and improving affect. The mechanisms through which exercise can accomplish this goal have been the subject of substantial research. CONCLUSIONS: Exercise is safe for individuals with back pain, because it does not increase the risk of future back injuries or work absence. Substantial evidence exists supporting the use of exercise as a therapeutic tool to improve impairments in back flexibility and strength. Most studies have observed improvements in global pain ratings after exercise programs, and many have observed that exercise can lessen the behavioral, cognitive, affect and disability aspects of back pain syndromes. PMID- 14749200 TI - Diastematomyelia presenting as progressive weakness in an adult after spinal fusion for adolescent idiopathic scoliosis. AB - BACKGROUND CONTEXT: Diastematomyelia is uncommon and rarely presents in adulthood. This report draws attention to the fact that patients who underwent spinal fusion for deformity before the widespread use of computed tomography (CT) and magnetic resonance imaging (MRI) may have unrecognized spinal cord abnormalities. This should be considered if revision surgery is contemplated. PURPOSE: This case report focuses on the late presentation of lower-extremity weakness in a 44-year-old woman with a split cord malformation (diplomyelia), diastematomyelia and tethered cord syndrome. STUDY DESIGN/SETTING: METHODS: The patient underwent instrumented posterior spinal fusion with a Harrington rod as a child for progressive thoracolumbar scoliosis. As an adult, she developed paraparesis after a traumatic event. The patient underwent decompressive laminectomy, subtotal resection of the old fusion mass and resection of the osseous septum. Postoperatively, an anterior spinal fluid leak in the lower thoracic region required repeated fascial grafting, resection of a pseudomeningocele and reverse left latissimus dorsi flap transfer. The leak was controlled, and the patient had near complete resolution of her paraparesis 1 year after her surgery. RESULTS: The case described herein is unusual in that patients with diplomyelia and diastematomyelia rarely are symptomatic in adulthood. However, trauma may precipitate the onset of neurologic symptoms. This patient underwent spinal surgeries to address deformity, pain and progressive lower-extremity weakness. Preoperative CT and MRI studies showed a split cord malformation and diastematomyelia at L1-L2 with spinal stenosis and tethering of both hemicords. CONCLUSIONS: Progressive weakness without any previous neurologic deficit or neurocutaneous stigmas of an underlying spinal cord abnormality may develop in the adult with unrecognized diastemotomyelia. This case demonstrates that a thorough preoperative workup of patients with complex spinal deformities is imperative. PMID- 14749203 TI - Insulin action during late pregnancy in the conscious dog. AB - Our aim was to assess the magnitude of peripheral insulin resistance and whether changes in hepatic insulin action were evident in a canine model of late (3rd trimester) pregnancy. A 3-h hyperinsulinemic (5 mU.kg(-1).min(-1)) euglycemic clamp was conducted using conscious, 18-h-fasted pregnant (P; n = 6) and nonpregnant (NP; n = 6) female dogs in which catheters for intraportal insulin infusion and assessment of hepatic substrate balances were implanted approximately 17 days before experimentation. Arterial plasma insulin rose from 11 +/- 2 to 192 +/- 24 and 4 +/- 2 to 178 +/- 5 microU/ml in the 3rd h in NP and P, respectively. Glucagon fell equivalently in both groups. Basal net hepatic glucose output was lower in NP (1.9 +/- 0.1 vs. 2.4 +/- 0.2 mg.kg(-1).min(-1), P < 0.05). Hyperinsulinemia completely suppressed hepatic glucose release in both groups (-0.4 +/- 0.2 and -0.1 +/- 0.2 mg.kg(-1).min(-1) in NP and P, respectively). More exogenous glucose was required to maintain euglycemia in NP (15.2 +/- 1.3 vs. 11.5 +/- 1.1 mg.kg(-1).min(-1), P < 0.05). Nonesterified fatty acids fell similarly in both groups. Net hepatic gluconeogenic amino acid uptake with high insulin did not differ in NP and P. Peripheral insulin action is markedly impaired in this canine model of pregnancy, whereas hepatic glucose production is completely suppressed by high circulating insulin levels. PMID- 14749204 TI - Distinct pathways regulate facilitated glucose transport in human articular chondrocytes during anabolic and catabolic responses. AB - Articular cartilage is an avascular, non-insulin-sensitive tissue that utilizes glucose as the main energy source, a precursor for glycosaminoglycan synthesis, and a regulator of gene expression. Facilitated glucose transport represents the first rate-limiting step in glucose metabolism. Previously, we demonstrated that glucose transport in chondrocytes is regulated by proinflammatory cytokines via upregulation of GLUT mRNA and protein expression. The objective of the present study was to determine differences in molecular mechanisms regulating glucose transport in chondrocytes stimulated with the anabolic transforming growth factor beta1 (TGF-beta1) vs. the catabolic and proinflammatory cytokine IL-1beta. Both TGF-beta1 and IL-1beta accelerate glucose transport in chondrocytes. Although both IL-1beta and TGF-beta1 enhance glucose transport in chondrocytes to a similar magnitude, IL-1beta induces significantly higher levels of lactate. TGF beta1-stimulated glucose transport is not associated with increased expression or membrane incorporation of GLUT1, -3, -6, -8, and -10 and depends on PKC and ERK activation. In contrast, IL-1beta-stimulated glucose transport is accompanied by increased expression and membrane incorporation of GLUT1 and -6 and depends upon activation of PKC and p38 MAP kinase. In conclusion, anabolic and catabolic stimuli regulate facilitated glucose transport in human articular chondrocytes via different effector and signaling mechanisms, and they have distinct effects on glycolysis. PMID- 14749205 TI - Catabolic action of insulin in rat arcuate nucleus is not enhanced by exogenous "tub" expression. AB - The central nervous system (CNS) protein "tub" has been identified from the genetically obese "tubby" mouse. Although the native function of tub in situ is not understood, cell-based studies suggest that one of its roles may be as an intracellular signaling target for insulin. In normal animals, insulin acts at the hypothalamic arcuate nucleus (ARC) to regulate energy balance. Here we used a Herpes Simplex viral expression system to evaluate whether tub overexpression in the ARC of normal rats enhances this action of insulin. In chow-fed rats, tub overexpression had no effect on insulin action. In rats fed a high-fat diet snack in addition to chow, simulating the diet of Westernized societies, the body weight regulatory action of insulin was impaired, and tub overexpression further impaired insulin action. Thus an excess of tub at the ARC does not enhance the in vivo effectiveness of insulin and is not able to compensate for the "downstream" consequences of a high-fat diet to impair CNS body weight regulatory mechanisms. PMID- 14749206 TI - Effects of rosiglitazone on gene expression in subcutaneous adipose tissue in highly active antiretroviral therapy-associated lipodystrophy. AB - Highly active antiretroviral therapy (HAART) has improved the prognosis of human immunodeficiency virus (HIV)-infected patients but is associated with severe adverse events, such as lipodystrophy and insulin resistance. Rosiglitazone did not increase subcutaneous fat in patients with HAART-associated lipodystrophy (HAL) in a randomized, double-blind, placebo-controlled trial, although it attenuated insulin resistance and decreased liver fat content. The aim of this study was to examine effects of rosiglitazone on gene expression in subcutaneous adipose tissue in 30 patients with HAL. The mRNA concentrations in subcutaneous adipose tissue were measured using real-time PCR. Twenty-four-week treatment with rosiglitazone (8 mg/day) compared with placebo significantly increased the expression of adiponectin, peroxisome proliferator-activated receptor-gamma (PPARgamma), and PPARgamma coactivator 1 and decreased IL-6 expression. Expression of other genes involved in lipogenesis, fatty acid metabolism, or glucose transport, such as acyl-CoA synthase, adipocyte lipid-binding protein, CD45, fatty acid transport protein-1 and -4, GLUT1, GLUT4, keratinocyte lipid binding protein, lipoprotein lipase, PPARdelta, and sterol regulatory element binding protein-1c, remained unchanged. Rosiglitazone also significantly increased serum adiponectin concentration. The change in serum adiponectin concentration was inversely correlated with the change in fasting serum insulin concentration and liver fat content. In conclusion, rosiglitazone induced significant changes in gene expression in subcutaneous adipose tissue and ameliorated insulin resistance in patients with HAL. Increased expression of adiponectin might have mediated most of the favorable insulin-sensitizing effects of rosiglitazone in these patients. PMID- 14749207 TI - Protein kinase Ciota enhances the transcriptional activity of the porcine P-450 side-chain cleavage insulin-like response element. AB - IGF-I enhances steroidogenesis in granulosa cells by stimulating the expression of the rate-limiting steroidogenic enzyme, cytochrome P-450 side-chain cleavage (P-450(scc)). This effect is mediated through an IGF response element (IGFRE) that binds polypyrimidine tract-binding protein (PTB)-associated splicing factor (PSF) and Sp1. Sp1 is essential for activation of the IGFRE, and PSF functions as a repressor. We investigated mechanisms of modulation of the IGFRE by the atypical protein kinase C (PKC)iota in a porcine stable granulosa cell line, JC 410. PKCiota was found in nuclear extracts, and levels were increased by IGF-I after 24 and 48 h of treatment. Immunoprecipitation experiments demonstrated that PSF and PKCiota associated with each other in nuclear extracts from JC-410 cells. Transient transfection with expression plasmids of kinase-active and kinase deficient PKCiota isoforms enhanced transcriptional activity of the IGFRE regardless of kinase catalytic activity. Depletion of PKCiota protein by small interfering RNA suppressed basal IGFRE activity but did not prevent IGF-I stimulation of the IGFRE. We conclude that PKCiota enhances transcriptional activity of the porcine P-450(scc) IGFRE independently of kinase activity by a mechanism involving protein-protein interaction with PSF. PMID- 14749208 TI - Reduced plasma FFA availability increases net triacylglycerol degradation, but not GPAT or HSL activity, in human skeletal muscle. AB - Intramuscular triacylglycerols (IMTG) are proposed to be an important metabolic substrate for contracting muscle, although this remains controversial. To test the hypothesis that reduced plasma free fatty acid (FFA) availability would increase IMTG degradation during exercise, seven active men cycled for 180 min at 60% peak pulmonary O(2) uptake either without (CON) or with (NA) prior ingestion of nicotinic acid to suppress adipose tissue lipolysis. Skeletal muscle and adipose tissue biopsy samples were obtained before and at 90 and 180 min of exercise. NA ingestion decreased (P < 0.05) plasma FFA at rest and completely suppressed the exercise-induced increase in plasma FFA (180 min: CON, 1.42 +/- 0.07; NA, 0.10 +/- 0.01 mM). The decreased plasma FFA during NA was associated with decreased (P < 0.05) adipose tissue hormone-sensitive lipase (HSL) activity (CON: 13.9 +/- 2.5, NA: 9.1 +/- 3.0 nmol.min(-1).mg protein(-1)). NA ingestion resulted in decreased whole body fat oxidation and increased carbohydrate oxidation. Despite the decreased whole body fat oxidation, net IMTG degradation was greater in NA compared with CON (net change: CON, 2.3 +/- 0.8; NA, 6.3 +/- 1.2 mmol/kg dry mass). The increased IMTG degradation did not appear to be due to reduced fatty acid esterification, because glycerol 3-phosphate activity was not different between trials and was unaffected by exercise (rest: 0.21 +/- 0.07; 180 min: 0.17 +/- 0.04 nmol.min(-1).mg protein(-1)). HSL activity was not increased from resting rates during exercise in either trial despite elevated plasma epinephrine, decreased plasma insulin, and increased ERK1/2 phosphorylation. AMP activated protein kinase (AMPK)alpha1 activity was not affected by exercise or NA, whereas AMPKalpha2 activity was increased (P < 0.05) from rest during exercise in NA and was greater (P < 0.05) than in CON at 180 min. These data suggest that plasma FFA availability is an important mediator of net IMTG degradation, and in the absence of plasma FFA, IMTG degradation cannot maintain total fat oxidation. These changes in IMTG degradation appear to disassociate, however, from the activity of the key enzymes responsible for synthesis and degradation of this substrate. PMID- 14749209 TI - Adipose depot-specific modulation of angiotensinogen gene expression in diet induced obesity. AB - Adipose tissue represents an important source of angiotensinogen (AGT). We investigated the effect of obesity induced by a high-fat diet on the expression of mouse (mAGT) and human AGT (hAGT) genes in liver, kidney, and heart and different adipose depots in normal mice (C57BL/6J), and in transgenic mice expressing the hAGT gene under the control of its own promoter. Mice were fed a high-fat diet (45% kcal) or normal chow (10% kcal) for 10 and 20 wk. The expression of mAGT and hAGT mRNA was quantified using an RNAse protection assay. Mice on the high-fat diet exhibited increased weight, fat mass, and plasma leptin. Expression of mAGT or hAGT genes was not affected by high-fat diet in nonadipose tissues, brown adipose tissue, or subcutaneous white fat. In contrast, high-fat diet increased both mAGT and hAGT gene expression in visceral adipose depots (omental, reproductive, and perirenal fat). Thus obesity-induced by a high fat diet is associated with a tissue-specific increased expression of both mouse and human AGT genes in intra-abdominal adipose tissue. Our findings also suggest that 1.2 kb of regulatory sequences present in the hAGT transgene are sufficient to transcriptionally respond to a high-fat diet in an adipose-specific and depot specific manner. PMID- 14749210 TI - IGF-I/IGFBP-3 ameliorates alterations in protein synthesis, eIF4E availability, and myostatin in alcohol-fed rats. AB - Chronic alcohol consumption decreases the concentration of the anabolic hormone IGF-I, and this change is associated with impaired muscle protein synthesis. The present study evaluated the ability of IGF-I complexed with IGF-binding protein (IGFBP)-3 to modulate the alcohol-induced inhibition of muscle protein synthesis in gastrocnemius. After 16 wk on an alcohol-containing diet, either the IGF I/IGFBP-3 binary complex (BC) or saline was injected two times daily for three consecutive days. After the final injection of BC (3 h), plasma IGF-I concentrations were elevated in alcohol-fed rats to values not different from those of similarly treated control animals. Alcohol feeding decreased the basal rate of muscle protein synthesis by limiting translational efficiency. BC treatment of alcohol-fed rats increased protein synthesis back to basal control values, but the rate remained lower than that of BC-injected control rats. The BC partially reversed the alcohol-induced decrease in the binding of eukaryotic initiation factor (eIF)4E with eIF4G. This change was associated with reversal of the alcohol-induced dephosphorylation of eIF4G but was independent of changes in the phosphorylation of either 4E-BP1 or eIF4E. However, BC reversed the alcohol induced increase in IGFBP-1 and muscle myostatin, known negative regulators of IGF-I action and muscle mass. Hence, exogenous IGF-I, administered as part of a BC to increase its circulating half-life, can in part reverse the decreased protein synthesis observed in muscle from chronic alcohol-fed rats by stimulating selected components of translation initiation. The data support the role of IGF-I as a mediator of chronic alcohol myopathy in rats. PMID- 14749212 TI - PKC-delta and CaMKII-delta 2 mediate ATP-dependent activation of ERK1/2 in vascular smooth muscle. AB - ATP, a purinergic receptor agonist, has been shown to be involved in vascular smooth muscle (VSM) cell DNA synthesis and cell proliferation during embryonic and postnatal development, after injury, and in atherosclerosis. One mechanism that ATP utilizes to regulate cellular function is through activation of ERK1/2. In the present study, we provide evidence that ATP-dependent activation of ERK1/2 in VSM cells utilizes specific isoforms of the multifunctional serine/threonine kinases, PKC, and Ca(2+)/calmodulin-dependent protein kinase II (CaMKII) as intermediates. Selective inhibition of PKC-delta activity with rottlerin, or adenoviral overexpression of kinase-negative PKC-delta, attenuated the ATP- and phorbol 12,13-dibutyrate (PDBu)-stimulated ERK1/2 activation. Inhibition of PKC alpha activity with Go-6976, or adenoviral overexpression of kinase-negative PKC alpha, was ineffective. Alternatively, treatment with KN-93, a selective inhibitor of CaMKII activation, or adenoviral overexpression of kinase-negative CaMKII-delta(2), inhibited ATP-dependent activation of ERK1/2 but had no effect on PDBu- or PDGF-stimulated ERK1/2. In addition, adenoviral overexpression of dominant-negative ras (Ad.HA-Ras(N17)) partially inhibited the ATP- and PDBu induced activation of ERK1/2 and blocked ionomycin- and EGF-stimulated ERK1/2, and inhibition of tyrosine kinases with AG-1478, an EGFR inhibitor, or the src family kinase inhibitor PP2 attenuated ATP-stimulated ERK1/2 activation. Taken together, these data indicate that PKC-delta and CaMKII-delta(2) coordinately mediate ATP-dependent transactivation of EGF receptor, resulting in increased ERK1/2 activity in VSM cells. PMID- 14749211 TI - Involvement of the mitogen-activated protein kinase cascade in peroxynitrite mediated arachidonic acid release in vascular smooth muscle cells. AB - Eicosanoid production is reduced when the nitric oxide (NO.) pathway is inhibited or when the inducible NO synthase gene is deleted, indicating that the NO. and arachidonic acid pathways are linked. We hypothesized that peroxynitrite, formed by the reaction of NO. and superoxide anion, may cause signaling events leading to arachidonic acid release and subsequent eicosanoid generation. Western blot analysis of rat arterial smooth muscle cells demonstrated that peroxynitrite (100 500 microM) and 3-morpholinosydnonimine (SIN-1; 200 microM) stimulate phosphorylation of extracellular signal-regulated kinase (ERK), p38, and cytosolic phospholipase A(2) (cPLA(2)). We found that peroxynitrite-induced arachidonic acid release was completely abrogated by the mitogen-activated protein/ERK kinase (MEK) inhibitor U0126 and by calcium chelators. With the p38 inhibitor SB-20219, we demonstrated that peroxynitrite-induced p38 phosphorylation led to minor arachidonic acid release, whereas U0126 completely blocked p38 phosphorylation. Addition of arachidonic acid caused p38 phosphorylation, suggesting that arachidonic acid or its metabolites are responsible for p38 activation. KN-93, a specific inhibitor of Ca(2+)/calmodulin dependent kinase II (CaMKII), revealed no role for this kinase in peroxynitrite induced arachidonic acid release in our cell system. Together, these results show that in response to peroxynitrite the cell initiates the MEK/ERK cascade leading to cPLA(2) activation and arachidonic acid release. Thus studies investigating the role of the NO. pathway on eicosanoid production must consider the contribution of signaling pathways initiated by reactive nitrogen species. These findings may provide evidence for a new role of peroxynitrite as an important reactive nitrogen species in vascular disease. PMID- 14749213 TI - Identification of the beta-subunit for nongastric H-K-ATPase in rat anterior prostate. AB - The structural organization of nongastric H-K-ATPase, unlike that of closely related Na-K-ATPase and gastric H-K-ATPase, is not well characterized. Recently, we demonstrated that nongastric H-K-ATPase alpha-subunit (alpha(ng)) is expressed in apical membranes of rodent prostate. Its highest level, as well as relative abundance, with respect to alpha(1)-isoform of Na-K-ATPase, was observed in anterior lobe. Here, we aimed to determine the subunit composition of nongastric H-K-ATPase through the detailed analysis of the expression of all known X-K ATPase beta-subunits in rat anterior prostate (AP). RT-PCR detects transcripts of beta-subunits of Na-K-ATPase only. Measurement of absolute protein content of these three beta-subunit isoforms, with the use of quantitative Western blotting of AP membrane proteins, indicates that the abundance order is beta(1) > beta(3) >> beta(2). Immunohistochemical experiments demonstrate that beta(1) is present predominantly in apical membranes, coinciding with alpha(ng), whereas beta(3) is localized in the basolateral compartment, coinciding with alpha(1). This is the first direct demonstration of the alpha(ng)-beta(1) colocalization in situ indicating that, in rat AP, alpha(ng) associates only with beta(1). The existence of alpha(ng-)beta(1) complex has been confirmed by immunoprecipitation experiments. These results indicate that beta(1)-isoform functions as the authentic subunit of Na-K-ATPase and nongastric H-K-ATPase. Putatively, the intracellular polarization of X-K-ATPase isoforms depends on interaction with other proteins. PMID- 14749215 TI - Tropomyosin interacts with phosphorylated HSP27 in agonist-induced contraction of smooth muscle. AB - Displacement of the contractile protein tropomyosin from actin filament exposes the myosin-binding sites on actin, resulting in actin-myosin interaction and muscle contraction. The objective of the present study was to better understand the interaction of tropomyosin with heat shock protein (HSP)27 in contraction of smooth muscle cells of the colon. We investigated the possibility of a direct protein-protein interaction of tropomyosin with HSP27 and the role of phosphorylated HSP27 in this interaction. Immunoprecipitation studies on rabbit smooth muscle cells indicate that upon acetylcholine-induced contraction tropomyosin shows increased association with HSP27 phosphorylated at Ser82 and Ser78. Transfection of smooth muscle cells with HSP27 phosphorylation mutants indicated that the association of tropomyosin with HSP27 could be affected by HSP27 phosphorylation. In vitro binding studies with glutathione S-transferase (GST)-tagged HSP27 mutant proteins show that tropomyosin has greater direct interaction to phosphomimic HSP27 mutant compared with wild-type and nonphosphomimic HSP27. Our data suggest that, in response to a contractile agonist, HSP27 undergoes a rapid phosphorylation that may strengthen its interaction with tropomyosin. PMID- 14749214 TI - Perinuclear localization of Na-K-Cl-cotransporter protein after human cytomegalovirus infection. AB - We (41) previously reported that Na-K-Cl-cotransporter (NKCC) function and microsomal protein expression are both dramatically reduced late in human cytomegalovirus (HCMV) infection of a human fibroblast cell line (MRC-5). We now report DNA microarray data showing that no significant HCMV-dependent NKCC gene repression can be detected 30 h postexposure (PE) to the virus. Consequently, we used plasma membrane biotinylation and subsequent subcellular fractionation in combination with semiquantitative immunoblotting and confocal microscopy to investigate the possibility that intracellular redistribution of the NKCC protein after HCMV infection could be a cause of the HCMV-induced loss of NKCC ion transport function. Our results show that the lifetime of plasmalemmal NKCC protein in quiescent, uninfected MRC-5 cells is approximately 48 h, and <20% of the total expressed NKCC protein are in the plasma membrane. The remainder (approximately 80%) was detected as diffusely distributed, small punctate structures in the cytoplasm. Following HCMV infection: 1) NKCC protein expression in the plasmalemma was sharply reduced (approximately 75%) within 24 h PE and thereafter continued to slowly decrease; 2) total cellular NKCC protein content remained unchanged or slightly increased during the course of the viral infection; and 3) HCMV infection caused NKCC protein to accumulate in the perinuclear region late in the HCMV infection (72 h PE). Thus our results imply that, in the process of productive HCMV infection, NKCC protein continues to be synthesized, but, instead of being delivered to the plasma membrane, it is clustered in a large, detergent-soluble perinuclear structure. PMID- 14749216 TI - Non-cross-bridge calcium-dependent stiffness in frog muscle fibers. AB - At the end of the force transient elicited by a fast stretch applied to an activated frog muscle fiber, the force settles to a steady level exceeding the isometric level preceding the stretch. We showed previously that this excess of tension, referred to as "static tension," is due to the elongation of some elastic sarcomere structure, outside the cross bridges. The stiffness of this structure, "static stiffness," increased upon stimulation following a time course well distinct from tension and roughly similar to intracellular Ca(2+) concentration. In the experiments reported here, we investigated the possible role of Ca(2+) in static stiffness by comparing static stiffness measurements in the presence of Ca(2+) release inhibitors (D600, Dantrolene, (2)H(2)O) and cross bridge formation inhibitors [2,3-butanedione monoxime (BDM), hypertonicity]. Both series of agents inhibited tension; however, only D600, Dantrolene, and (2)H(2)O decreased at the same time static stiffness, whereas BDM and hypertonicity left static stiffness unaltered. These results indicate that Ca(2+), in addition to promoting cross-bridge formation, increases the stiffness of an (unidentified) elastic structure of the sarcomere. This stiffness increase may help in maintaining the sarcomere length uniformity under conditions of instability. PMID- 14749217 TI - Urea transport in MDCK cells that are stably transfected with UT-A1. AB - Progress in understanding the cell biology of urea transporter proteins has been hampered by the lack of an appropriate cell culture system. The goal of this study was to create a polarized epithelial cell line that stably expresses the largest of the rat renal urea transporter UT-A isoforms, UT-A1. The gene for UT A1 was cloned into pcDNA5/FRT and transfected into Madin-Darby canine kidney (MDCK) cells with an integrated Flp recombination target site. The cells from a single clone were grown to confluence on collagen-coated membranes until the resistance was >1,500 Omega.cm(2). Transepithelial [(14)C]urea fluxes were measured at 37 degrees C in a HCO(3)(-)/CO(2) buffer, pH 7.4, with 5 mM urea. The baseline fluxes were not different between unstimulated UT-A1-transfected MDCK cells and nontransfected or sham-transfected MDCK cells. However, only in the UT A1-transfected cells was UT-A1 protein expressed (as measured by Western blot analysis) and urea transport stimulated by forskolin or arginine vasopressin. Forskolin and arginine vasopressin also increased the phosphorylation of UT-A1. Thionicotinamide, dimethylurea, and phloretin inhibited the forskolin-stimulated [(14)C]urea fluxes in the UT-A1-transfected MDCK cells. These characteristics mimic those seen in rat terminal inner medullary collecting ducts. This new polarized epithelial cell line stably expresses UT-A1 and reproduces several of the physiological responses observed in rat terminal inner medullary collecting ducts. PMID- 14749219 TI - Protein synthesis rates of human PBMC and PMN can be determined simultaneously in vivo by using small blood samples. AB - Immune cell functions can be evaluated in vivo by measuring their specific protein fractional synthesis rates (FSR). Using stable isotope dilution techniques, we describe a new method allowing simultaneous in vivo assessment of FSR in two leukocyte populations in healthy human subjects, using small blood samples. Peripheral blood mononuclear cell (PBMC) and polymorphonuclear neutrophil (PMN) FSR were measured during primed continuous intravenous infusion of l-[1-(13)C]leucine. Immune cells from 6 ml of whole blood were isolated by density gradient centrifugation. In a first study, we calculated the FSR using plasma [(13)C]leucine or alpha-[(13)C]ketoisocaproate (KIC) enrichments as precursor pools. In a second study, we compared protein FSR in leukocytes, using enrichments of either intracellular or plasma free [(13)C]leucine as immediate precursor pools. The present approach showed a steady-state enrichment of plasma and circulating immune cell free [(13)C]leucine precursor pools. The linearity of labeled amino acid incorporation rate within mixed PBMC and PMN proteins also was verified. Postabsorptive protein FSR was 4.09 +/- 0.39%/day in PBMC and 1.44 +/- 0.08%/day in PMN when plasma [(13)C]KIC was the precursor pool. The difference between PBMC and PMN FSR was statistically significant, whatever the precursor pool used, suggesting large differences in their synthetic activities and functions. Use of the plasma [(13)C]KIC pool led to an underestimation of leukocyte FSR compared with the intracellular pool (PBMC: 6.04 +/- 0.94%/day; PMN: 2.98 +/- 0.30%/day). Hence, the intracellular free amino acid pool must be used as precursor to obtain reliable results. In conclusion, it is possible to assess immune cell metabolism in vivo in humans by using small blood samples to directly indicate their metabolic activity in various clinical situations and in response to regulating factors. PMID- 14749220 TI - Energy expenditure in obesity. PMID- 14749221 TI - Phytoestrogens and breast cancer. PMID- 14749222 TI - Renal metabolism of amino acids: its role in interorgan amino acid exchange. AB - The kidneys play a role in the synthesis and interorgan exchange of several amino acids. The quantitative importance of renal amino acid metabolism in the body is not, however, clear. We review here the role of the kidney in the interorgan exchange of amino acids, with emphasis on quantitative aspects. We reviewed relevant literature by using a computerized literature search (PubMed) and checking relevant references from the identified articles. Our own data are discussed in the context of the literature. The kidney takes up glutamine and metabolizes it to ammonia. This process is sensitive to pH and serves to maintain acid-base homeostasis and to excrete nitrogen. In this way, the metabolism of renal glutamine and ammonia is complementary to hepatic urea synthesis. Citrulline, derived from intestinal glutamine breakdown, is converted to arginine by the kidney. Renal phenylalanine uptake is followed by stoichiometric tyrosine release, and glycine uptake is accompanied by serine release. Certain administered oligopeptides (eg, glutamine dipeptides) are converted by the kidneys to their constituent components before they can be used in metabolic processes. The kidneys play an important role in the interorgan exchange of amino acids. Quantitatively, for several important amino acids, the kidneys are as important as the gut in intermediary metabolism. The kidneys may be crucial "mediators" of the beneficial effects of specialized, disease-specific feeding solutions such as those enriched in glutamine dipeptides. PMID- 14749223 TI - Nutrition education of medical and dental students: innovation through curriculum integration. AB - Nutrition is a necessary component of education in the health professions. Although often underplayed, nutrition is an integral facet of dental education, particularly because the oral cavity is the entry point to the gastrointestinal tract. This article addresses the current status of nutrition education in medical and dental schools, including the common themes, strategies, and challenges of integrating nutrition education in this venue, particularly in dental schools. The survival and progression of nutrition as a component of medical and dental education depends to a large extent on the creativity and innovative strategies used by educators and administrators in medical and dental schools and in training programs. A forward-thinking attitude with a focus on the integration of nutrition topics throughout the 4 y of medical or dental school and subsequent training programs will increase the potential for a successful program. PMID- 14749224 TI - Effects of moderate-fat (from monounsaturated fat) and low-fat weight-loss diets on the serum lipid profile in overweight and obese men and women. AB - BACKGROUND: Little evidence of the effects of moderate-fat (from monounsaturated fat) weight-loss diets on risk factors for cardiovascular disease exists because low-fat diets are typically recommended. Previous studies in weight-stable persons showed that a moderate-fat diet results in a more favorable lipid and lipoprotein profile (ie, lower serum triacylglycerol and higher HDL cholesterol) than does a low-fat diet. OBJECTIVE: We evaluated the effects of energy controlled, low-fat and moderate-fat diets on changes in lipids and lipoproteins during weight loss and subsequent weight maintenance. DESIGN: We conducted a parallel-arm study design in overweight and obese [body mass index (in kg/m(2)): 29.8 +/- 2.4] healthy men and women (n = 53) assigned to consume a low-fat (18% of energy) or moderate-fat (33% of energy) diet for 6 wk to achieve weight loss, which was followed by 4 wk of weight maintenance. All foods were provided and body weight was monitored to ensure equal weight loss between groups. RESULTS: The moderate-fat diet elicited favorable changes in the lipoprotein profile. Compared with baseline, HDL cholesterol was unchanged, whereas triacylglycerol and the ratios of total and non-HDL cholesterol to HDL cholesterol were lower at the end of the weight-maintenance period in the moderate-fat diet group. Despite similar weight loss, triacylglycerol rebounded, HDL cholesterol decreased, and the ratios of total and non-HDL cholesterol to HDL cholesterol did not change during the 10-wk interval in the low-fat diet group. CONCLUSIONS: A moderate-fat weight-loss and weight-maintenance diet improves the cardiovascular disease risk profile on the basis of favorable changes in lipids and lipoproteins. There is merit in recommending a moderate-fat weight-loss diet. PMID- 14749225 TI - Fruit and vegetable consumption and LDL cholesterol: the National Heart, Lung, and Blood Institute Family Heart Study. AB - BACKGROUND: An elevated LDL-cholesterol concentration is associated with an increased risk of cardiovascular disease. The association between fruit and vegetable consumption and LDL has been inconsistent. OBJECTIVE: The objective was to determine whether a high intake of fruit and vegetables is inversely associated with LDL concentrations. DESIGN: We used data collected from 4466 subjects in the National Heart, Lung, and Blood Institute Family Heart Study to study the association between fruit and vegetable consumption and serum LDL. We used a food-frequency questionnaire to assess fruit and vegetable intakes and regression models to estimate adjusted mean LDL according to fruit and vegetable consumption. RESULTS: The mean (+/-SD) age of the men (n = 2047) was 51.5 +/- 14.0 y and that of the women (n = 2419) was 52.2 +/- 13.7 y. The average daily serving of fruit and vegetables was 3.2 +/- 1.7 for men and was 3.5 +/- 1.8 for women. Fruit and vegetable consumption was inversely related to LDL: in the categories 0-1.9, 2.0-2.9, 3.0-3.9, and > or = 4 servings/d, multivariate adjusted mean (95% CI) LDL concentrations were 3.36 (3.28, 3.44), 3.35 (3.27, 3.43), 3.26 (3.17, 3.35), and 3.17 (3.09, 3.25) mmol/L, respectively, for men (P for trend < 0.0001) and 3.35 (3.26, 3.44), 3.22 (3.14, 3.30), 3.21 (3.13, 3.29), and 3.11 (3.04, 3.18), respectively, for women (P for trend < 0.0001). This association was observed across categories of age, education, smoking status, physical activity, and tertiles of Keys score. Exclusion of subjects with prevalent diabetes mellitus or coronary artery disease did not alter these results significantly. CONCLUSION: Consumption of fruit and vegetables is inversely related to LDL in men and women. PMID- 14749226 TI - Use of serum retinol-binding protein for prediction of vitamin A deficiency: effects of HIV-1 infection, protein malnutrition, and the acute phase response. AB - BACKGROUND: Serum retinol is the most commonly used indicator of vitamin A status. Retinol is transported in a 1-to-1 complex with retinol-binding protein (RBP). RBP is easy and inexpensive to measure, and studies have shown a high correlation between concentrations of RBP and concentrations of retinol. The performance of RBP in the context of infection or protein malnutrition, however, has not been evaluated. OBJECTIVE: Our aim was to determine whether RBP is a good surrogate measure for retinol in the context of HIV-1 infection, protein malnutrition, and the acute phase response. DESIGN: The relation between RBP and retinol was examined in a cross-sectional study of 600 Kenyan women. RESULTS: There was a high correlation between concentrations of RBP and those of retinol (r = 0.88). When equimolar cutoffs were used, RBP predicted marginal vitamin A status (retinol < 1.05 micro mol/L) with 93% sensitivity and 75% specificity and vitamin A deficiency (retinol < 0.70 micro mol/L) with 91% sensitivity and 94% specificity. Similarly high sensitivities and specificities were found among subgroups with HIV-1 infection, a positive acute phase response, and protein malnutrition. Protein malnutrition and a positive acute phase response were common, especially among HIV-1-infected women, and were independently and synergistically associated with lower RBP concentrations. CONCLUSIONS: Equimolar RBP cutoffs predict vitamin A deficiency with high sensitivity and specificity, even in the context of infection and protein malnutrition. Like retinol, RBP may not accurately identify true vitamin A status under all conditions, because the acute phase response and protein malnutrition depress RBP concentrations. However, RBP may be a simple, inexpensive tool for assessment of vitamin A deficiency in population studies. PMID- 14749227 TI - Oxidative stress in abetalipoproteinemia patients receiving long-term vitamin E and vitamin A supplementation. AB - BACKGROUND: Patients with abetalipoproteinemia develop progressive ataxic neuropathy and retinopathy that are thought to be due, in part, to oxidative damage resulting from deficiencies of vitamins E and A. OBJECTIVE: The goal was to determine the degree of oxidative stress in abetalipoproteinemia patients who had received vitamin E (100 mg/kg) and vitamin A (10 000-15 000 IU/d) since infancy. DESIGN: Ten patients aged 3-25 y were studied. Assessed were plasma carbonyl concentrations as a marker of oxidative damage to proteins; total plasma oxidizability, which was used to evaluate the susceptibility of plasma lipoproteins to oxidation; and cyclic voltammetry, which represents the overall reducing and antioxidant capacity stemming from low-molecular-weight antioxidants in plasma. RESULTS: Concentrations of plasma carbonyls did not differ significantly between patients and control subjects ( +/- SE: 0.5670 +/- 0.031 and 0.5039 +/- 0.0134 nmol/mg protein, respectively). The lag phase of plasma oxidizability was 28.03 +/- 3.16 min in the patients and 24.0 +/- 2.79 min in healthy subjects in whom oxidizability of isolated HDL was measured (NS). Cyclic voltammetry showed a peak potential of 330 +/- 8.3 mV in all samples studied, denoting that the same antioxidants were present in the plasma of the patients and the control subjects. The anodic current of the samples, a measure of the concentration of hydrophilic low-molecular-weight antioxidants, was 5.227 +/- 0.25 and 5.38 +/- 0.20 micro A in the patients and the control subjects, respectively (NS). CONCLUSION: Enhanced oxidative stress is not apparent in the plasma of abetalipoproteinemia patients receiving long-term supplementation with vitamins E and A. PMID- 14749228 TI - New reference values for thyroid volume by ultrasound in iodine-sufficient schoolchildren: a World Health Organization/Nutrition for Health and Development Iodine Deficiency Study Group Report. AB - BACKGROUND: Goiter prevalence in school-age children is an indicator of the severity of iodine deficiency disorders (IDDs) in a population. In areas of mild to-moderate IDDs, measurement of thyroid volume (Tvol) by ultrasound is preferable to palpation for grading goiter, but interpretation requires reference criteria from iodine-sufficient children. OBJECTIVE: The study aim was to establish international reference values for Tvol by ultrasound in 6-12-y-old children that could be used to define goiter in the context of IDD monitoring. DESIGN: Tvol was measured by ultrasound in 6-12-y-old children living in areas of long-term iodine sufficiency in North and South America, central Europe, the eastern Mediterranean, Africa, and the western Pacific. Measurements were made by 2 experienced examiners using validated techniques. Data were log transformed, used to calculate percentiles on the basis of the Gaussian distribution, and then transformed back to the linear scale. Age- and body surface area (BSA)-specific 97th percentiles for Tvol were calculated for boys and girls. RESULTS: The sample included 3529 children evenly divided between boys and girls at each year ( +/- SD age: 9.3 +/- 1.9 y). The range of median urinary iodine concentrations for the 6 study sites was 118-288 micro g/L. There were significant differences in age- and BSA-adjusted mean Tvols between sites, which suggests that population specific references in countries with long-standing iodine sufficiency may be more accurate than is a single international reference. However, overall differences in age- and BSA-adjusted Tvols between sites were modest relative to the population and measurement variability, which supports the use of a single, site-independent set of references. CONCLUSION: These new international reference values for Tvol by ultrasound can be used for goiter screening in the context of IDD monitoring. PMID- 14749229 TI - Effects of biotin on pyruvate carboxylase, acetyl-CoA carboxylase, propionyl-CoA carboxylase, and markers for glucose and lipid homeostasis in type 2 diabetic patients and nondiabetic subjects. AB - BACKGROUND: Several studies have shown that biotin affects glucose homeostasis. Serum biotin concentrations are lower in subjects with type 2 diabetes than in control subjects. Lymphocyte propionyl-CoA carboxylase (PCC; EC 6.4.1.3) activity has proved to be a sensitive indicator of biotin status that is more accurate than is serum biotin concentration. OBJECTIVE: We studied the activity of PCC, pyruvate carboxylase (PC; EC 6.4.1.1), and acetyl-CoA carboxylase (ACC; EC 6.4.1.2) in type 2 diabetic and nondiabetic subjects. The effect of biotin administration (6.14 micro mol/d) on the activity of these enzymes and on several plasma metabolites was also studied. DESIGN: We compared the activities of carboxylases in circulating lymphocytes from patients with type 2 diabetes (n = 24) with those in circulating lymphocytes from nondiabetic subjects (n = 30). We also assessed the effect of biotin administration for 14 and 28 d on the activity of these enzymes and on the concentrations of several metabolites (type 2 diabetic patients, n = 10; nondiabetic subjects, n = 7). RESULTS: No significant differences in lymphocyte carboxylase activities were found between the type 2 diabetic patients and the nondiabetic subjects. Biotin administration increased the activity of PCC, PC, and ACC in all the subjects. No significant change in glucose, insulin, triacylglycerol, cholesterol, or lactate concentration was observed with the treatment in either the diabetic or the nondiabetic subjects. CONCLUSIONS: The activity of carboxylases does not differ significantly between type 2 diabetic and nondiabetic subjects. Pharmacologic doses of biotin increase lymphocyte PCC, PC, and ACC activities. PMID- 14749230 TI - Effect of a micronutrient fortificant mixture and 2 amounts of calcium on iron and zinc absorption from a processed food supplement. AB - BACKGROUND: Iron, zinc, and calcium can interact with each other in a way that inhibits their respective absorption. On the other hand, mineral fortification has been used to improve simultaneous iron and zinc absorption from food supplements. OBJECTIVE: We evaluated the effect of a novel fortificant mixture consisting of NaFeEDTA, zinc methionine, ascorbic acid, and citric acid on iron and zinc absorption from a dry food supplement designed for preschool children. DESIGN: The standard food supplement contained cereal and legume flour, dried milk, and a mixture of micronutrients including ferrous sulfate and zinc sulfate as sources of supplemental iron and zinc, respectively. Standard and novel food products were prepared as porridge with or without the addition of 200 mg Ca as calcium phosphate. Iron absorption and zinc absorption from the food products were evaluated simultaneously in 13 nonpregnant, adult women by extrinsically labeling the products with radioisotopes of iron and zinc and carrying out whole body counting 7 d after the food products were consumed in random order. RESULTS: The absorption of iron from the NaFeEDTA-containing (novel) food product was 1.7 times that from the ferrous sulfate-containing (standard) product (P = 0.015). There was no significant effect of dietary calcium on iron absorption. Zinc absorption was not associated with the form of zinc consumed, but higher dietary calcium was marginally associated with lower zinc absorption (P = 0.071). CONCLUSIONS: A mixture of fortificants containing NaFeEDTA, zinc sulfate or zinc methionine, ascorbic acid, and citric acid, but without calcium, can improve iron and zinc absorption from food products. A cost-benefit analysis of the novel fortificant mixture needs to be performed. PMID- 14749231 TI - Effect of alpha-linolenic acid supplementation during pregnancy on maternal and neonatal polyunsaturated fatty acid status and pregnancy outcome. AB - BACKGROUND: Maternal essential fatty acid status declines during pregnancy, and as a result, neonatal concentrations of docosahexaenoic acid (DHA, 22:6n-3) and arachidonic acid (AA, 20:4n-6) may not be optimal. OBJECTIVE: Our objective was to improve maternal and neonatal fatty acid status by supplementing pregnant women with a combination of alpha-linolenic acid (ALA, 18:3n-3) and linoleic acid (LA, 18:2n-6), the ultimate dietary precursors of DHA and AA, respectively. DESIGN: From week 14 of gestation until delivery, pregnant women consumed daily 25 g margarine supplying either 2.8 g ALA + 9.0 g LA (n = 29) or 10.9 g LA (n = 29). Venous blood was collected for plasma phospholipid fatty acid analyses at weeks 14, 26, and 36 of pregnancy, at delivery, and at 32 wk postpartum. Umbilical cord blood and vascular tissue samples were collected to study neonatal fatty acid status also. Pregnancy outcome variables were assessed. RESULTS: ALA+LA supplementation did not prevent decreases in maternal DHA and AA concentrations during pregnancy and, compared with LA supplementation, did not increase maternal and neonatal DHA concentrations but significantly increased eicosapentaenoic acid (20:5n-3) and docosapentaenoic acid (22:5n-3) concentrations. In addition, ALA+LA supplementation lowered neonatal AA status. No significant differences in pregnancy outcome variables were found. CONCLUSIONS: Maternal ALA+LA supplementation did not promote neonatal DHA+AA status. The lower concentrations of Osbond acid (22:5n-6) in maternal plasma phospholipids and umbilical arterial wall phospholipids with ALA+LA supplementation than with LA supplementation suggest only that functional DHA status improves with ALA+LA supplementation. PMID- 14749232 TI - Long-term consumption of infant formulas containing live probiotic bacteria: tolerance and safety. AB - BACKGROUND: Nonpathogenic live bacteria are consumed as food by many children, particularly in the form of yogurt. The tolerance and safety of long-term consumption of specific types and strains of probiotic bacteria are not well documented. OBJECTIVE: The goal was to evaluate tolerance to formulas containing 2 levels of probiotic supplementation and effects on growth, general clinical status, and intestinal health in free-living healthy infants. DESIGN: This was a prospective, double-blind, randomized, placebo-controlled study of healthy infants aged 3-24 mo. Infants were assigned to receive a standard milk-based formula containing 1 x 10(7) colony-forming units (CFU)/g each of Bifidobacterium lactis and Streptococcus thermophilus, formula containing 1 x 10(6) CFU/g each of B. lactis and S. thermophilus, or unsupplemented formula. Clinical outcomes included formula intake, gastrointestinal tolerance, anthropometric measures, daycare attendance, and history of illness. RESULTS: One hundred eighteen infants aged ( +/- SD) 7.0 +/- 2.9 mo at enrollment consumed formula for 210 +/- 127 d. There were no significant differences in age, sex, formula consumption, or length of study between groups. The supplemented formulas were well accepted and were associated with a lower frequency of reported colic or irritability (P < 0.001) and a lower frequency of antibiotic use (P < 0.001) than was the unsupplemented formula. There were no significant differences between groups in growth, health care attention seeking, daycare absenteeism, or other health variables. CONCLUSION: Long-term consumption of formulas supplemented with B. lactis and S. thermophilus was well tolerated and safe and resulted in adequate growth, reduced reporting of colic or irritability, and a lower frequency of antibiotic use. PMID- 14749233 TI - Energy expenditure in African American and white boys and girls in a 2-y follow up of the Baton Rouge Children's Study. AB - BACKGROUND: Previously reported race and sex differences in energy expenditure (EE) may play a role in body fat gain. OBJECTIVE: The purpose of the study was to determine the relations between race, sex, Tanner stage, and EE. DESIGN: We conducted a 2-y follow-up study of EE in 114 African American (AA) and white girls and boys aged 12.7 +/- 0.1 y ( +/- SE), who were stratified as obese or lean and were part of the Baton Rouge Children's Study. Total daily EE (TDEE) was measured by using doubly labeled water. Resting metabolic rate (RMR) and thermic effect of food were measured by using indirect calorimetry. RESULTS: White children had significantly higher TDEE and RMR than did AA children when fat-free mass was considered. Boys had significantly higher TDEE and RMR than did girls, even after adjustment for differences in size. TDEE and RMR were significantly higher in obese children, as a result of their greater fat-free mass and body fat, than in lean children. Activity-related EE did not differ significantly between obese and lean children. There was a strong relation between initial and 2-y TDEE and RMR. There was a significant decrease in activity-related EE in both racial groups. AA children had significantly more lean limb mass than did white children. CONCLUSIONS: Average TDEE did not change over 2 y, but RMR increased significantly, and activity-related EE decreased significantly. Differences in trunk and limb lean mass of white and AA children may explain some of the ethnic differences in EE. The decrease in physical activity over 2 y may contribute to the risk of obesity. PMID- 14749234 TI - Plant foods, fiber, and rectal cancer. AB - BACKGROUND: Associations between colon and rectal cancer and intakes of vegetables, other plant foods, and fiber have stimulated much debate. OBJECTIVE: We examined the association between rectal cancer and plant food and fiber intakes. DESIGN: Data from 952 incident cases of rectal cancer were compared with data from 1205 population-based controls living in Utah or enrolled in the Kaiser Permanente Medical Care Program in northern California RESULTS: Rectal cancer was inversely associated with intakes of vegetables (odds ratio: 0.72; 95% CI: 0.54, 0.98), fruit (0.73; 0.53, 0.99), and whole-grain products (0.69; 0.51, 0.94), whereas a high intake of refined-grain products was directly associated with an increased risk of rectal cancer (1.42; 1.04, 1.92). Similarly, relative to low fiber intakes, high intakes of dietary fiber reduced the risk of rectal cancer (0.54; 0.37, 0.78). The reduced risk of rectal cancer associated with vegetable (0.48; 0.29, 0.80), fruit (0.63; 0.38, 1.06), and fiber (0.40; 0.22, 0.71) intakes was strongest for persons who received the diagnosis after age 65 y. A threshold effect at approximately 5 servings of vegetables/d was needed to see a reduced risk of rectal cancer. CONCLUSIONS: The results suggest that plant foods may be important in the etiology of rectal cancer in both men and women. Age at diagnosis appears to play an important role in the association. PMID- 14749235 TI - Dietary phytoestrogens and breast cancer risk. AB - BACKGROUND: A high intake of phytoestrogens, particularly isoflavones, has been suggested to decrease breast cancer risk. Results from human studies are inconclusive. OBJECTIVE: We investigated the association between phytoestrogen intake and breast cancer risk in a large prospective study in a Dutch population with a habitually low phytoestrogen intake. DESIGN: The study population consisted of 15 555 women aged 49-70 y who constituted a Dutch cohort of the European Prospective Investigation into Cancer and Nutriton (EPIC; 1993-1997). Data concerning habitual dietary intake in the preceding year were obtained by using a validated food-frequency questionnaire. The content of isoflavones and lignans in relevant food items was estimated through a literature search, the use of food-composition tables, and contact with experts. Newly diagnosed breast cancer cases up to 1 January 2001 were identified through linkage with the Comprehensive Cancer Center Middle Netherlands. Hazard ratios for the disease were estimated by Cox proportional hazard analysis for quartiles of isoflavone and lignan intake. Associations were adjusted for known breast cancer risk factors and daily energy intake. RESULTS: A total of 280 women were newly diagnosed with breast cancer during follow-up. The median daily intakes of isoflavones and lignans were 0.4 (interquartile range: 0.3-0.5) and 0.7 (0.5-0.8) mg/d, respectively. Relative to the respective lowest intake quartiles, the hazard ratios for the highest intake quartiles for isoflavones and lignans were 1.0 (95% CI: 0.7, 1.5) and 0.7 (0.5, 1.1), respectively. Tests for trend were nonsignificant. CONCLUSION: In Western populations, a high intake of isoflavones or mammalian lignans is not significantly related to breast cancer risk. PMID- 14749236 TI - Antioxidants and physical performance in elderly persons: the Invecchiare in Chianti (InCHIANTI) study. AB - BACKGROUND: Muscle strength and physical performance in old age might be related to the oxidative damage caused by free radicals. OBJECTIVE: The objective was to assess the correlation of plasma concentrations and daily dietary intakes of antioxidants with skeletal muscle strength and physical performance in elderly persons. DESIGN: This study is part of the Invecchiare in Chianti (InCHIANTI) study, which was conducted in 986 Italians aged > or = 65 y. Physical performance was assessed on the basis of walking speed, ability to rise from a chair, and standing balance. Knee extension strength was assessed with a hand-held dynamometer. The European Prospective Investigation into Cancer and Nutrition (EPIC) questionnaire was used to evaluate the daily dietary intakes of vitamin C, vitamin E, beta-carotene, and retinol. Plasma alpha- and gamma-tocopherol concentrations were measured. Adjusted linear regression analyses were used to calculate regression coefficients per SD increase in plasma concentrations and daily dietary intakes. RESULTS: In adjusted analyses, plasma alpha-tocopherol was significantly correlated with knee extension (beta = 0.566, P = 0.003) and the summary physical performance score (beta = 0.044, P = 0.008). Plasma gamma tocopherol was associated only with knee extension strength (beta = 0.327, P = 0.04). Of the daily dietary intake measures, vitamin C and beta-carotene were significantly correlated with knee extension strength, and vitamin C was significantly associated with physical performance (beta = 0.029, P = 0.04). CONCLUSIONS: Plasma antioxidant concentrations correlate positively with physical performance and strength. Higher dietary intakes of most antioxidants, especially vitamin C, appear to be associated with higher skeletal muscular strength in elderly persons. PMID- 14749237 TI - Edentulism and nutritional status in a biracial sample of well-functioning, community-dwelling elderly: the health, aging, and body composition study. AB - BACKGROUND: Edentulism may affect dietary intake in older adults, but the relation between edentulism and nutritional status is not completely understood. OBJECTIVE: The present study examined whether edentulism is associated with nutritional status and whether there is an interaction between race and edentulism on nutritional status among well-functioning, community-dwelling elderly. DESIGN: The study cohort included 3075 elderly aged 70-79 y (52% women, 42% black) in the Health, Aging, and Body Composition Study. Dietary intake, anthropometric variables, weight change, and serum albumin and lipid concentrations were compared between edentate and dentate participants by the use of multiple linear and logistic regressions. RESULTS: Edentulism was not associated with total energy or food intake but was associated with the food groups consumed, particularly fat, micronutrients, and hard-to-chew foods. Edentulism was more strongly linked to dietary intake in whites than in blacks. Unlike black edentate elderly, white edentate elderly consumed significantly lower energy-adjusted amounts of vitamin A and beta-carotene, higher amounts of energy-adjusted total and saturated fat and cholesterol, and higher percentages of energy from fat than did white dentate elderly. Anthropometry and biochemical indexes were not significantly different by edentulism status in both races. Edentulism was associated with weight gains of >5% in 1 y in both races. CONCLUSIONS: Edentulism was associated with differences in the nutritional status of well-functioning, community-dwelling elderly, more so in whites than blacks. Edentate elders may benefit from dental, medical, and nutrition interventions targeted to addressing these findings. PMID- 14749239 TI - Fruit and vegetable intakes are an independent predictor of bone size in early pubertal children. AB - BACKGROUND: Adequate intakes of fruit and vegetables are recommended for optimum health in children. OBJECTIVE: The objective of this study was to determine whether consuming fruit and vegetables >3 times per day is beneficial to bone mass in children. DESIGN: Fifty-six white females (Tanner stage 2) recorded dietary intake on 3 independent days. The numbers of servings of fruit and vegetables were recorded for each day and tallied, and the subjects were divided into 2 consumption groups for analysis (low consumption: <3 servings/d, n = 22; high consumption: > or = 3 servings/d, n = 34). Bone area and the bone mineral content of the whole body and radius were assessed by using dual-energy X-ray absorptiometry. Radioimmunoassays measured serum parathyroid hormone and 25 hydroxyvitamin D. Twenty-four-hour urine samples were assessed for calcium, sodium, and creatinine. RESULTS: After adjustment for age, body mass index, and physical activity, those children who reported consuming > or = 3 servings fruit and vegetables/d had more bone area of the whole body (6.0%; P = 0.03) and radius (8.3%; P = 0.03), lower urinary calcium excretion (2.6 +/- 0.2 compared with 1.8 +/- 0.3 mg/kg; P = 0.04), and lower parathyroid hormone (19.6 +/- 1.9 compared with 25.0 +/- 1.6 pg/mL; P = 0.01) than did those children who reported consuming <3 servings fruit and vegetables/d. CONCLUSIONS: High fruit and vegetable intakes have beneficial effects on the bone area of the radius and whole body in early pubertal girls. The lower urinary calcium output associated with higher fruit and vegetable intakes may be a modulating factor. PMID- 14749238 TI - Energy requirements in the eighth decade of life. AB - BACKGROUND: Knowledge of energy requirements among relatively healthy elderly is limited. OBJECTIVES: The objectives of the study were to measure total energy expenditure (TEE)-derived energy requirements in a biracial population of older adults without limitations to daily life and to test these empirical measures against national and international recommendations. DESIGN: TEE (measured by the doubly labeled water method), resting metabolic rate (RMR), activity-related energy expenditure (AEE), and body composition were measured in 288 persons aged 70-79 y selected from the Health, Aging, and Body Composition Study. RESULTS: TEE was lower in women (approximately 530 kcal/d; P < 0.0001) than in men because of the women's lower RMR and AEE. Fat-free mass explained the sex difference in RMR, but body weight failed to account for the women's lower AEE (approximately 1 kcal x kg(-1) x d(-1); P = 0.007). Blacks had lower TEE than did whites (approximately 100 kcal/d, P = 0.03), and that was explained by blacks' lower RMR. Physical activity level (TEE/RMR) did not differ significantly between sexes and races (1.70 +/- 0.23). The World Health Organization (WHO) recommendations overestimated TEE by 10 +/- 15% (P < 0.0001) in women but not in men, and the dietary reference intakes (DRIs) were accurate to 0 +/- 14% (P = 0.1). Both WHO and DRI recommendations are based on an underestimated physical activity level, and WHO recommendations are based on overestimated RMR. CONCLUSIONS: This study of well-functioning older adults confirms the racial difference in energy metabolism and supports the use of the 2002 DRIs. Because the DRIs and WHO recommendations underestimated PAL, new predictive equations of energy requirements are proposed. PMID- 14749240 TI - Supplementation with flaxseed alters estrogen metabolism in postmenopausal women to a greater extent than does supplementation with an equal amount of soy. AB - BACKGROUND: Phytoestrogens, which are abundant in flaxseed and soy, have chemical structures resembling those of endogenous estrogens and have been shown to exert hormonal effects, thereby affecting chronic diseases. OBJECTIVE: We compared the effects of consuming equal amounts of flaxseed or soy on estrogen metabolism and biochemical markers of bone metabolism in postmenopausal women. DESIGN: In a parallel design, the diet of postmenopausal women (n = 46) was supplemented with either a placebo, soy (25 g soy flour), or flaxseed (25 g ground flaxseed) muffin for 16 wk. Blood and 24-h urine samples were collected at baseline and at the endpoint. Urine samples were analyzed for phytoestrogens, estrogen metabolites (2 hydroxyestrone, 16alpha-hydroxyestrone), and serum hormones (estradiol, estrone, estrone sulfate). Serum and urine samples were also analyzed for biochemical markers of bone metabolism. RESULTS: Urinary concentrations of 2-hydroxyestrone, but not of 16alpha-hydroxyestrone, increased significantly in the flaxseed group (P = 0.05). In the flaxseed group, the ratio of 2-hydroxyestrone to 16alpha hydroxyestrone was positively correlated with urinary lignan excretion (r = 0.579, P = 0.02). In the soy and placebo groups, no significant correlation was observed. No significant change in serum hormones or biochemical markers of bone metabolism was observed within or between the treatment groups. CONCLUSIONS: Supplementation with flaxseed modifies urinary estrogen metabolite excretion to a greater extent than does supplementation with an equal amount of soy. This modification by flaxseed is associated with an increase in urinary lignan excretion. Despite the shift in estrogen metabolism to favor the less biologically active estrogens, a negative effect on bone cell metabolism was not observed. PMID- 14749241 TI - The effects of phytoestrogen isoflavones on bone density in women: a double blind, randomized, placebo-controlled trial. AB - BACKGROUND: Isoflavone phytoestrogen therapy has been proposed as a natural alternative to hormone replacement therapy (HRT). HRT has a beneficial effect on bone, but few trials in humans have investigated the effects of isoflavones on bone. OBJECTIVE: The objective of the study was to determine the effect on bone density of a red clover-derived isoflavone supplement that provided a daily dose of 26 mg biochanin A, 16 mg formononetin, 1 mg genistein, and 0.5 mg daidzein for 1 y. Effects on biochemical markers of bone turnover and body composition were also studied. DESIGN: Women aged 49-65 y (n = 205) were enrolled in a double blind, randomized, placebo-controlled trial; 177 completed the trial. Bone density, body composition, bone turnover markers, and diet were measured at baseline and after 12 mo. RESULTS: Loss of lumbar spine bone mineral content and bone mineral density was significantly (P = 0.04 and P = 0.03, respectively) lower in the women taking the isoflavone supplement than in those taking the placebo. There were no significant treatment effects on hip bone mineral content or bone mineral density, markers of bone resorption, or body composition, but bone formation markers were significantly increased (P = 0.04 and P = 0.01 for bone-specific alkaline phosphatase and N-propeptide of collagen type I, respectively) in the intervention group compared with placebo in postmenopausal women. Interactions between treatment group and menopausal status with respect to changes in other outcomes were not significant. CONCLUSION: These data suggest that, through attenuation of bone loss, isoflavones have a potentially protective effect on the lumbar spine in women. PMID- 14749243 TI - Sleep patterns, docosahexaenoic acid, and gestational length. PMID- 14749244 TI - Bioelectrical impedance analysis for predicting body composition: what about the external validity of new regression equations? PMID- 14749247 TI - Vitamin B-12 and folate deficiency in elderly persons. PMID- 14749248 TI - Dietary glycemic load, overall glycemic index, and serum insulin concentrations in healthy schoolchildren. PMID- 14749250 TI - Assessment of resting energy expenditure in mechanically ventilated patients. PMID- 14749249 TI - Limitations of plasma plant sterols as indicators of cholesterol absorption. PMID- 14749251 TI - Measuring bone mass accumulation. PMID- 14749252 TI - The function of the superficial root mat in the biogeochemical cycles of nutrients in congolese eucalyptus plantations. AB - BACKGROUND AND AIMS: The importance of superficial root mats inside the forest floor for the nutrition of Amazonian rain forests has been extensively investigated. The present study was aimed at assessing the function of a root mat adherent to decomposing organic material observed in Eucalyptus plantations. METHODS: The development of the root mat was studied through micromorphological observations of thin litter sections, and the influence of soil microtopography and soil water repellency on root mat biomass was assessed in situ on an area of 5 m2. In addition, input-output budgets of nutrients within the forest floor were established from measurements of litterfall, dissolved nutrients in gravitational solutions, and forest floor nutrient contents. KEY FINDINGS: The amounts of nutrients released during litter decay in this ecosystem during the period of study were, on average, 46, 3, 4, 19 and 17 kg ha-1 year-1 for N, P, K, Ca and Mg, respectively. The simultaneous measurements of the chemical composition of throughfall solutions and leachates beneath the forest floor showed a very quick uptake of nutrients by the root mat during the decomposition processes. Indeed, the solutions did not become noticeably enriched in nutrients during their passage through the holorganic layer, despite large amounts of elements being released during litter decay. The root mat biomass decreased significantly during the dry season, and a preferential development in microdepressions at the soil surface was observed. A strong water repellency observed in these depressions might enhance the ability of the roots to take up water and nutrients during the dry periods. CONCLUSIONS: The root mat was active throughout the year to catch the flux of nutrients from the biodegradation of the forest floor, preventing the transfer of dissolved nutrients toward deeper soil horizons. This mechanism is involved in the successful adaptation of this Eucalyptus hybrid in areas covered by 'climacic' savannas in Congo. PMID- 14749253 TI - Shoot development in grapevine (Vitis vinifera) is affected by the modular branching pattern of the stem and intra- and inter-shoot trophic competition. AB - BACKGROUND AND AIMS: Shoot architecture variability in grapevine (Vitis vinifera) was analysed using a generic modelling approach based on thermal time developed for annual herbaceous species. The analysis of shoot architecture was based on various levels of shoot organization, including pre-existing and newly formed parts of the stem, and on the modular structure of the stem, which consists of a repeated succession of three phytomers (P0-P1-P2). METHODS: Four experiments were carried out using the cultivar 'Grenache N': two on potted vines (one of which was carried out in a glasshouse) and two on mature vines in a vineyard. These experiments resulted in a broad diversity of environmental conditions, but none of the plants experienced soil water deficit. KEY RESULTS: Development of the main axis was highly dependent on air temperature, being linearly related to thermal time for all stages of leaf development from budbreak to veraison. The stable progression of developmental stages along the main stem resulted in a thermal-time based programme of leaf development. Leaf expansion rate varied with trophic competition (shoot and cluster loads) and environmental conditions (solar radiation, VPD), accounting for differences in final leaf area. Branching pattern was highly variable. Classification of the branches according to ternary modular structure increased the accuracy of the quantitative analysis of branch development. The rate and duration of leaf production were higher for branches derived from P0 phytomers than for branches derived from P1 or P2 phytomers. Rates of leaf production, expressed as a -function of thermal time, were not stable and depended on trophic competition and environmental conditions such as solar radiation or VPD. CONCLUSIONS: The application to grapevine of a generic model developed in annual plants made it possible to identify constants in main stem development and to determine the hierarchical structure of branches with respect to the modular structure of the stem in response to intra- and inter shoot trophic competition. PMID- 14749254 TI - An N-terminal peptide extension results in efficient expression, but not secretion, of a synthetic horseradish peroxidase gene in transgenic tobacco. AB - BACKGROUND AND AIMS: Native horseradish (Armoracia rusticana) peroxidase, HRP (EC 1.11.1.7), isoenzyme C is synthesized with N-terminal and C-terminal peptide extensions, believed to be associated with protein targeting. This study aimed to explore the specific functions of these extensions, and to generate transgenic plants with expression patterns suitable for exploring the role of peroxidase in plant development and defence. METHODS: Transgenic Nicotiana tabacum (tobacco) plants expressing different versions of a synthetic horseradish peroxidase, HRP, isoenzyme C gene were constructed. The gene was engineered to include additional sequences coding for either the natural N-terminal or the C-terminal extension or both. These constructs were placed under the control of a constitutive promoter (CaMV-35S) or the tobacco RUBISCO-SSU light inducible promoter (SSU) and introduced into tobacco using Agrobacterium-mediated transformation. To study the effects of the N- and C-terminal extensions, the localization of recombinant peroxidase was determined using biochemical and molecular techniques. KEY RESULTS: Transgenic tobacco plants can exhibit a ten-fold increase in peroxidase activity compared with wild-type tobacco levels, and the majority of this activity is located in the symplast. The N-terminal extension is essential for the production of high levels of recombinant protein, while the C-terminal extension has little effect. Differences in levels of enzyme activity and recombinant protein are reflected in transcript levels. CONCLUSIONS: There is no evidence to support either preferential secretion or vacuolar targeting of recombinant peroxidase in this heterologous expression system. This leads us to question the postulated targeting roles of these peptide extensions. The N terminal extension is essential for high level expression and appears to influence transcript stability or translational efficiency. Plants have been generated with greatly elevated cytosolic peroxidase activity, and smaller increases in apoplastic activity. These will be valuable for exploring the role of these enzymes in stress amelioration and plant development. PMID- 14749255 TI - Macula densa basolateral ATP release is regulated by luminal [NaCl] and dietary salt intake. AB - One component of the macula densa (MD) tubuloglomerular feedback (TGF) signaling pathway may involve basolateral release of ATP through a maxi-anion channel. Release of ATP has previously been studied during a maximal luminal NaCl concentration ([NaCl](L)) stimulus (20-150 mmol/l). Whether MD ATP release occurs during changes in [NaCl](L) within the physiological range (20-60 mmol/l) has not been examined. Also, because TGF is known to be enhanced by low dietary salt intake, we examined the pattern of MD ATP release from salt-restricted rabbits. Fluorescence microscopy, with fura 2-loaded cultured mouse mesangial cells as biosensors, was used to assess ATP release from the isolated, perfused thick ascending limb containing the MD segment. The mesangial biosensor cells, which contain purinergic receptors and elevate intracellular Ca(2+) concentration ([Ca(2+)](i)) on ATP binding, were placed adjacent to the MD basolateral membrane. Elevations in [NaCl](L) between 0 and 80 mmol/l, in 20-mmol/l increments, caused stepwise increases in [Ca(2+)](i), with the highest increase at [NaCl](L) of approximately 60 mmol/l. Luminal furosemide at 10(-4) mol/l blocked ATP release, which suggests that the efflux of ATP required MD Na-2Cl-K cotransport. A low-salt diet for 1 wk increased the magnitude of [NaCl](L) dependent elevations in biosensor [Ca(2+)](i) by twofold, whereas high-salt intake had no effect. In summary, ATP release occurs over the same range of [NaCl](L) (20-60 mmol/l) previously reported for TGF responses, and, similar to TGF, ATP release was enhanced by dietary salt restriction. Thus these two findings are consistent with the role of MD ATP release as a signaling component of the TGF pathway. PMID- 14749256 TI - Evidence for epidermal growth factor receptor as negative-feedback control in aldosterone-induced Na+ reabsorption. AB - Aldosterone enhances Na(+) reabsorption via epithelial Na(+) channels (ENaC). Aldosterone also stimulates the protein kinase ERK1/2- and the epidermal growth factor (EGF) receptor (EGFR)-signaling pathway. Yet EGF and ERK1/2 are known inhibitors of ENaC-mediated Na(+) reabsorption. In the present study, using the well-established Madin-Darby canine kidney C7 cell line, we tested the hypothesis that EGFR represents a negative-feedback control for chronic aldosterone-induced Na(+) reabsorption [amiloride-inhibitable short-circuit current (I(sc))]. Mineralocorticoid receptor expression was confirmed by RT-PCR and Western blot analysis. Aldosterone enhanced ERK1/2 phosphorylation in an EGFR-dependent way. Furthermore, aldosterone stimulated EGFR expression. Aldosterone (10 nmol/l) induced a small transient increase in I(sc) under control conditions. Inhibition of ERK1/2 phosphorylation with U-0126 (10 micromol/l) stimulated I(sc), indicating constitutive ENaC inhibition. Aldosterone exerted a significantly larger effect in the presence of U-0126 than without U-0126. EGF (10 microg/l) inhibited I(sc), whereas inhibition of EGFR kinase by tyrphostin AG-1478 (100 nmol/l) enhanced I(sc). Aldosterone was more effective in the presence of AG-1478 than without AG-1478. In summary, we propose that the EGFR-signaling cascade can serve as a negative-feedback control to limit the effect of aldosterone-induced Na(+) reabsorption. PMID- 14749257 TI - AE2 isoforms in rat kidney: immunohistochemical localization and regulation in response to chronic NH4Cl loading. AB - Three splice variants of anion exchanger (AE)2 (AE2a, b, and c) have been described in the rat, but their relative distribution in rat kidney is not known. The purpose of this study was to describe the segmental and cellular distribution of the AE2 isoforms in the rat kidney and to evaluate whether the expression levels of these AE2 isoforms are regulated independently in response to chronic NH(4)Cl loading. Two polyclonal antibodies were generated, respectively, recognizing a NH(2)-terminal peptide unique to AE2a and an amino acid sequence common to AE2a and AE2b. Antibody specificities were tested using cells transfected separately with the AE2a, AE2b, and AE2c isoforms. Immunohistochemistry on sections of paraffin-embedded rat kidneys showed a distribution of AE2a/AE2b labeling in the kidney similar to the distribution of AE2 in the rat kidney reported previously. AE2 is highly expressed in the medullary thick ascending limb, cortical thick ascending limb (cTAL), and macula densa. The pattern of AE2a-specific labeling differed from the pattern of AE2a/AE2b labeling in that relatively more of the total immunolabel was observed in the terminal inner medullary collecting duct. NH(4)Cl loading (0.033 mmol NH(4)Cl/g body wt for 7 days) did not change the labeling of AE2 isoforms in the medulla, whereas the labeling in the cortex was intensified and included more distal parts of the cTAL. Immunoblotting confirmed upregulation of AE2a/b expression in the cortex. These results indicate that AE2a and AE2b are differentially expressed and regulated in the rat kidney. The regulation following NH(4)Cl loading of AE2b in the cTAL suggests a role for AE2 in transepithelial bicarbonate reabsorption in this segment. PMID- 14749258 TI - Novel factors in the regulation of beta-cell function. PMID- 14749259 TI - Differential levels of diabetogenic stress in two new mouse models of obesity and type 2 diabetes. AB - The genetic basis for the more common forms of human obesity predisposing to insulin resistance and development of type 2 diabetes is multigenic rather than monogenic in origin. New mouse "diabesity" models have been created by combining independent diabetes risk-conferring quantitative trait loci from two unrelated parental strains: New Zealand Obese (NZO/HlLt) and Nonobese Nondiabetic (NON/Lt). F1 hybrid males, heterozygous at all polymorphic autosomal loci distinguishing the two parental strains, are driven to obesity-induced diabetes (diabesity) at high frequencies. This review focuses on two new recombinant congenic strains (RCSs) developed by introgressing multiple NZO/HlLt chromosomal segments into the nominally diabesity-resistant NON/Lt strain background. Both RCSs gain more weight than NON animals. Although exhibiting comparable weight gain and adiposity, only one of the two RCSs develops diabetes. Hence, these two RCSs will be instructive in elucidating genetic and pathophysiological differences underlying uncomplicated obesity syndromes versus diabetogenic obesity (diabesity) syndromes. Unlike mice with null mutations in a single gene producing morbid obesity, the new models develop a more moderate obesity produced by the interaction of numerous genes with relatively small effects. These RCSs are differentially sensitive to adverse side effects of thiazolidinediones and thus should be particularly useful for pharmacogenetic analyses. PMID- 14749260 TI - Structure, activation, and biology of calpain. AB - Variation in the calpain 10 gene has recently been shown to be associated with type 2 diabetes by positional cloning. Since then, studies on calpain 10 have been started in correlation with diabetes and insulin-mediated signaling. In this review, the activation mechanism of calpain by calcium ions, which is essential to understand its physiological functions, is discussed on the basis of recent X ray structural analyses. Further, special features of the structure of calpain 10 that differ from those of typical micro - or m-calpain used in most studies are summarized together with discussion of the physiological function of calpain with respect to type 2 diabetes. PMID- 14749261 TI - Linkage of calpain 10 to type 2 diabetes: the biological rationale. AB - The follow-up studies to the original report of association of variation at calpain 10 (CAPN10) with type 2 diabetes in the Mexican-American population of Starr County, Texas, encompass a broad range of science. There are association studies on genetic variation at CAPN10 in different human populations over a range of phenotypes related to type 2 diabetes, physiological studies on the biological functions of calpain proteases, and evolutionary studies on CAPN10 and the NIDDM1 region. We review here the studies published to date on CAPN10, as well as the latest findings from positional cloning studies on a number of other complex disorders. Collectively, these studies provide perspective on the challenges of moving from the linkage mapping and positional cloning studies on which we have been focused to an understanding of the biology shaping the relationship of genotype to phenotype at loci influencing susceptibility to complex disorders like type 2 diabetes. PMID- 14749262 TI - Genetic factors and insulin secretion: gene variants in the IGF genes. AB - IGFs are important regulators of pancreatic beta-cell development, growth, and maintenance. Mutations in the IGF genes have been found to be associated with type 2 diabetes, myocardial infarction, birth weight, and obesity. These associations could result from changes in insulin secretion. We have analyzed glucose-stimulated insulin secretion using hyperglycemic clamps in carriers of a CA repeat in the IGF-I promoter and an ApaI polymorphism in the IGF-II gene. Normal and impaired glucose-tolerant subjects (n = 237) were independently recruited from three different populations in the Netherlands and Germany to allow independent replication of associations. Both first- and second-phase insulin secretion were not significantly different between the various IGF-I or IGF-II genotypes. Remarkably, noncarriers of the IGF-I CA repeat allele had both a reduced insulin sensitivity index (ISI) and disposition index (DI), suggesting an altered balance between insulin secretion and insulin action. Other diabetes related parameters were not significantly different for both the IGF-I and IGF-II gene variant. We conclude that gene variants in the IGF-I and IGF-II genes are not associated with detectable variations in glucose-stimulated insulin secretion in these three independent populations. Further studies are needed to examine the exact contributions of the IGF-I CA repeat alleles to variations in ISI and DI. PMID- 14749263 TI - Single nucleotide polymorphisms in the proximal promoter region of the adiponectin (APM1) gene are associated with type 2 diabetes in Swedish caucasians. AB - Adiponectin (APM1) is an adipocyte-derived peptide. The APM1 gene is located on chromosome 3q27 and linked to type 2 diabetes. In patients with type 2 diabetes, the adiponectin level in plasma is decreased in comparison to healthy subjects. To identify genetic defects of the APM1 gene that contribute to the development of type 2 diabetes, we genotyped 13 single nucleotide polymorphisms (SNPs) in 106 patients with type 2 diabetes, 325 patients with impaired glucose tolerance (IGT), and 497 nondiabetic control subjects in Swedish Caucasians by using dynamic allele-specific hybridization (DASH). We found that SNPs -11426(A/G) and 11377(G/C) in the proximal promoter region had significant differences of allele frequencies between type 2 diabetic patients and nondiabetic control subjects (P = 0.02 and P = 0.04, respectively). SNP-11426(A/G) was significantly associated with fasting plasma glucose in type 2 diabetic patients (P = 0.02) and in IGT subjects (P = 0.04), while the patients carrying CC and CG genotypes for SNP 11377(G/C) had a higher BMI than the patients with the GG genotype (P = 0.03). Haplotype analysis of 13 SNPs in the APM1 gene showed that estimates of haplotype frequencies in Swedish Caucasians are similar to those estimated in French Caucasians. However, no significant association of haplotypes with type 2 diabetes and IGT was detected in our study. The present study provides additional evidence that SNPs in the proximal promoter region of the APM1 gene contribute to the development of type 2 diabetes. PMID- 14749264 TI - Putative metabolic effects of the liver X receptor (LXR). AB - The nuclear receptors liver X receptor (LXR)alpha and LXRbeta are sensors of cholesterol metabolism and lipid biosynthesis. They have recently been found to be regulators of inflammatory cytokines, suppressors of hepatic glucose production, and involved in different cell-signaling pathways. LXRalpha is a target gene of the peroxisome proliferator-activated receptor-gamma, a target of drugs used in treating elevated levels of glucose seen in diabetes. Furthermore, insulin induces LXRalpha in hepatocytes, resulting in increased expression of lipogenic enzymes and suppression of key enzymes in gluconeogenesis, including PEPCK. LXR seems to have an important role in the regulation of glucocorticoid action and a role in the overall energy homeostasis suggested by its putative regulatory effect on leptin and uncoupling protein 1. The physiological roles of LXR indicate that it is an interesting potential target for drug treatment of diabetes. PMID- 14749265 TI - The biology of peroxisome proliferator-activated receptors: relationship with lipid metabolism and insulin sensitivity. AB - Peroxisome proliferator-activated receptors (PPARs) are transcription factors belonging to the superfamily of nuclear receptors. Three isoforms (alpha, delta, and gamma) have been described. They act on DNA response elements as heterodimers with the nuclear retinoic acid receptor. Their natural activating ligands are fatty acids and lipid-derived substrates. PPAR-alpha is present in liver, heart, and, to a lesser extent, skeletal muscle. When activated, it promotes fatty acid oxidation, ketone body synthesis, and glucose sparing. Fibrates, which are used as hypolipidemic drugs, are ligands of PPAR-alpha. PPAR-delta is ubiquitous and could also favor fatty acid oxidation in tissues in which PPAR-alpha is absent or less expressed. PPAR-gamma is expressed in adipose tissue, lower intestine, and cells involved in immunity. Activation of PPAR-gamma induces the differentiation of preadipocytes into adipocytes and stimulates triglyceride storage. Thiazolidinediones are compounds used as hypoglycemic, muscle insulin-sensitizing agents in type 2 diabetes. Unexpectedly, they are activators of PPAR-gamma. Their action on muscle insulin sensitivity may be secondary to the lowering of circulating lipids on PPAR-gamma activation and to the secretion by adipocytes of insulin-sensitizing hormones such as adiponectin, all promoting glucose utilization. The PPARs are thus major regulators of lipid and glucose metabolism, allowing adaptation to the prevailing nutritional environment. PMID- 14749266 TI - Aged transgenic mice with increased glucocorticoid sensitivity in pancreatic beta cells develop diabetes. AB - Glucocorticoids are diabetogenic hormones because they decrease glucose uptake, increase hepatic glucose production, and inhibit insulin release. To study the long-term effects of increased glucocorticoid sensitivity in beta-cells, we studied transgenic mice overexpressing the rat glucocorticoid receptor targeted to the beta-cells using the rat insulin I promoter. Here we report that these mice developed hyperglycemia both in the fed and the overnight-fasted states at 12-15 months of age. Progression from impaired glucose tolerance, previously observed in the same colony at the age of 3 months, to manifest diabetes was not associated with morphological changes or increased apoptosis in the beta-cells. Instead, our current results suggest that the development of diabetes is due to augmented inhibition of insulin secretion through alpha(2)-adrenergic receptors (alpha(2)-ARs). Thus, we found a significantly higher density of alpha(2)-ARs in the islets of transgenic mice compared with controls, based on binding studies with the alpha(2)-AR agonist UK 14304. Furthermore, incubation of islets with benextramine, a selective antagonist of the alpha(2)-AR, restored insulin secretion in response to glucose in isolated islets from transgenic mice, whereas it had no effect on control islets. These results indicate that the chronic enhancement of glucocorticoid signaling in pancreatic beta-cells results in hyperglycemia and impaired glucose tolerance. This effect may involve signaling pathways that participate in the regulation of insulin secretion via the alpha(2) AR. PMID- 14749267 TI - Role of peroxisome proliferator-activated receptor-gamma in the glucose-sensing apparatus of liver and beta-cells. AB - Type 2 diabetes develops in the context of both insulin resistance and beta-cell failure. Thiazolidinediones are a class of antidiabetic agents that are known to improve insulin sensitivity in various animal models of diabetes. The improved insulin sensitivity may be achieved either by systemic insulin sensitization or by direct action of peroxisome proliferator-activated receptor (PPAR)-gamma on the transcription of genes involved in glucose disposal. Evidence supporting the direct action of PPAR-gamma on glucose metabolism is observed in the genes involved in insulin-stimulated glucose disposal. We already showed that GLUT2 and beta-glucokinase were directly activated by PPAR-gamma. Recently, we have identified and characterized the functional PPAR response element in the GLUT2 and liver type glucokinase (LGK) promoter of the liver. It is well known that adipose tissue plays a crucial role in antidiabetic action of PPAR-gamma. In addition, PPAR-gamma can directly affect liver and pancreatic beta-cells to improve glucose homeostasis. PMID- 14749268 TI - Liver glucokinase can be activated by peroxisome proliferator-activated receptor gamma. AB - Thiazolidinediones (TZDs), synthetic ligands of peroxisome proliferator-activated receptor (PPAR)-gamma, are known to decrease hepatic glucose production and increase glycogen synthesis in diabetic animals. Recently it was reported that glucokinase (GK) expression was increased by TZDs in the liver of diabetic ZDF rats. However, the mechanism whereby TZDs increase GK expression is not yet studied. We have assumed that liver type glucokinase (LGK) induction by TZDs could be achieved by direct transcriptional activation. Thus, we have dissected the LGK promoter to explore the presence of a PPAR response element (PPRE) in the promoter. From this study, we were able to localize a PPRE in the -116/-104 region of the rat LGK gene. The PPAR-gamma/retinoid X receptor-alpha heterodimer was bound to the element and activated the LGK promoter. The LGK promoter lacking the PPRE or having mutations in the PPRE could not be activated by PPAR-gamma. Furthermore, troglitazone increased endogenous GK mRNA in primary hepatocytes. These results indicate that PPAR-gamma can directly activate GK expression in liver and may contribute to improving glucose homeostasis in type 2 diabetes. PMID- 14749269 TI - Potential role of peroxisome proliferator-activated receptor-alpha in the modulation of glucose-stimulated insulin secretion. AB - In this review, we discuss the influence of peroxisome proliferator-activated receptor (PPAR)-alpha on islet insulin secretion and develop the hypothesis that modulation of PPAR-alpha function may be important for the regulation of compensatory insulin secretion. We have attempted to analyze the role of PPAR alpha-linked fatty acid metabolism in islet function in health and in insulin resistant states linked to lifestyle factors, in particular pregnancy and a diet inappropriately high in saturated fat. We have emphasized the potential for both actions of PPAR-alpha on insulin sensitivity that may be relayed systemically to the islet, leading to modulation of the insulin response in accordance with changes in insulin sensitivity, and direct effects of PPAR-alpha action on the islet itself. Finally, we have developed the concept that compensatory insulin secretion may have a function not only in glucoregulation but also in liporegulation. Thus, augmented insulin secretion may reflect a requirement for lipid lowering as well as for increased glucose disposal and is perceived to aim to compensate for impaired suppression of islet lipid delivery by insulin. This introduces the possibility of a continuum between liporegulation with islet compensation and lipodysregulation leading to islet decompensation in the development of type 2 diabetes. PMID- 14749270 TI - Nuclear protein p8 is associated with glucose-induced pancreatic beta-cell growth. AB - On its own, glucose is a major factor for proliferation of pancreatic beta-cells and is also an essential prerequisite for IGF-I and growth hormone-induced growth of these cells. p8 was originally identified as an emergency gene product upregulated in pancreatic acinar cells in response to acute pancreatitis. p8 was further shown to be involved in a broad range of biological functions, including cell growth, growth arrest, apoptosis, and tumor development. These in part opposite actions may be related to distinct stimuli and pathways in certain conditions and cell types. Here we demonstrate that p8 is widely expressed in human pancreatic islets in vivo and in several beta-cell lines in vitro. Based on this observation, we tested the hypothesis that p8 production in pancreatic beta cells is regulated by glucose. Incubation of rat INS-1 beta-cells with 25 mmol/l glucose resulted in a continuous increase of proliferating cell numbers. This was accompanied by a strong upregulation of p8 mRNA and protein expression, indicating that p8 is a physiological mediator of glucose-induced pancreatic beta cell growth. Binding of glucose-activated protein kinase C (PKC) to two PKC sites within a highly conserved region of the p8 protein may be a possible mechanism linking glucose and p8 pathways leading to proliferation. PMID- 14749271 TI - Novel players in pancreatic islet signaling: from membrane receptors to nuclear channels. AB - Glucose and other nutrients regulate many aspects of pancreatic islet physiology. This includes not only insulin release, but also insulin synthesis and storage and other aspects of beta-cell biology, including cell proliferation, apoptosis, differentiation, and gene expression. This implies that in addition to the well described signals for insulin release, other intracellular signaling mechanisms are needed. Here we describe the role of global and local Ca(2+) signals in insulin release, the regulation of these signals by new membrane receptors, and the generation of nuclear Ca(2+) signals involved in gene expression. An integrated view of these pathways should improve the present description of the beta-cell biology and provide new targets for novel drugs. PMID- 14749272 TI - Nutrients induce different Ca(2+) signals in cytosol and nucleus in pancreatic beta-cells. AB - Specific activation of Ca(2+)-dependent functions is achieved by the particular dynamics and local restriction of Ca(2+) signals. It has been shown that changes in amplitude, duration, or frequency of Ca(2+) signals modulate gene transcription. Thus, Ca(2+) variations should be finely controlled within the nucleus. Although a variety of mechanisms in the nuclear membrane have been demonstrated to regulate nuclear Ca(2+), the existence of an autonomous Ca(2+) homeostasis within the nucleus is still questioned. In the pancreatic beta-cell, besides their effect on insulin secretion, Ca(2+) messages generated by nutrients also exert their action on gene expression. However, the dynamics of these Ca(2+) signals in relation to nuclear function have been explored little in islet cells. In the current study, Ca(2+) changes both in the nucleoplasm and in the cytosol of INS-1 and pancreatic beta-cells were monitored using spot confocal microscopy. We show that nutrients trigger Ca(2+) signals of higher amplitude in the nucleus than in the cytosol. These amplitude-modulated Ca(2+) signals transmitted to the nucleus might play an important role in the control of gene expression in the pancreatic beta-cell. PMID- 14749273 TI - Roles of mitochondria in health and disease. AB - Mitochondria play a central role in cell life and cell death. An increasing number of studies place mitochondrial dysfunction at the heart of disease, most notably in the heart and the central nervous system. In this article, I review some of the key features of mitochondrial biology and focus on the pathways of mitochondrial calcium accumulation. Substantial evidence now suggests that the accumulation of calcium into mitochondria may play a key role as a trigger to mitochondrial pathology, especially when that calcium uptake is accompanied by another stressor, in particular nitrosative or oxidative stress. The major process involved is the opening of the mitochondrial permeability transition pore, a large conductance pore that causes a collapse of the mitochondrial membrane potential, leading to ATP depletion and necrotic cell death or to cytochrome c release and apoptosis, depending on the rate of ATP consumption. I discuss two models in particular in which these processes have been characterized. The first is a model of oxidative stress in cardiomyocytes, in which reperfusion after ischemia causes mitochondrial calcium overload, and oxidative stress. Recent experiments suggest that cardioprotection by hypoxic preconditioning or exposure to the ATP-dependent K(+) channel opener diazoxide increases mitochondrial resistance to oxidative injury. In a second model, of calcium overload in neurons, the neurotoxicity of glutamate depends on mitochondrial calcium uptake, but the toxicity to mitochondria also requires the generation of nitric oxide. Glutamate toxicity after activation of N-methyl-D aspartate (NMDA) receptors results from the colocalization of NMDA receptors with neuronal nitric oxide synthase (nNOS). The calcium increase mediated by NMDA receptor activation is thus associated with nitric oxide generation, and the combination leads to the collapse of mitochondrial membrane potential followed by cell death. PMID- 14749274 TI - Mitochondrial diabetes: molecular mechanisms and clinical presentation. AB - Mutations in mitochondrial DNA (mtDNA) associate with various disease states. A few mtDNA mutations strongly associate with diabetes, with the most common mutation being the A3243G mutation in the mitochondrial DNA-encoded tRNA(Leu,UUR) gene. This article describes clinical characteristics of mitochondrial diabetes and its molecular diagnosis. Furthermore, it outlines recent developments in the pathophysiological and molecular mechanisms leading to a diabetic state. A gradual development of pancreatic beta-cell dysfunction upon aging, rather than insulin resistance, is the main mechanism in developing glucose intolerance. Carriers of the A3243G mutation show during a hyperglycemic clamp at 10 mmol/l glucose a marked reduction in first- and second-phase insulin secretion compared with noncarriers. The molecular mechanism by which the A3243G mutation affects insulin secretion may involve an attenuation of cytosolic ADP/ATP levels leading to a resetting of the glucose sensor in the pancreatic beta-cell, such as in maturity-onset diabetes of the young (MODY)-2 patients with mutations in glucokinase. Unlike in MODY2, which is a nonprogressive form of diabetes, mitochondrial diabetes does show a pronounced age-dependent deterioration of pancreatic function indicating involvement of additional processes. Furthermore, one would expect that all mtDNA mutations that affect ATP synthesis lead to diabetes. This is in contrast to clinical observations. The origin of the age dependent deterioration of pancreatic function in carriers of the A3243G mutation and the contribution of ATP and other mitochondrion-derived factors such as reactive oxygen species to the development of diabetes is discussed. PMID- 14749275 TI - Prevention of mitochondrial oxidative damage as a therapeutic strategy in diabetes. AB - Hyperglycemia causes many of the pathological consequences of both type 1 and type 2 diabetes. Much of this damage is suggested to be a consequence of elevated production of reactive oxygen species by the mitochondrial respiratory chain during hyperglycemia. Mitochondrial radical production associated with hyperglycemia will also disrupt glucose-stimulated insulin secretion by pancreatic beta-cells, because pancreatic beta-cells are particularly susceptible to oxidative damage. Therefore, mitochondrial radical production in response to hyperglycemia contributes to both the progression and pathological complications of diabetes. Consequently, strategies to decrease mitochondrial radical production and oxidative damage may have therapeutic potential. This could be achieved by the use of antioxidants or by decreasing the mitochondrial membrane potential. Here, we outline the background to these strategies and discuss how antioxidants targeted to mitochondria, or selective mitochondrial uncoupling, may be potential therapies for diabetes. PMID- 14749276 TI - Beta-cell glucose toxicity, lipotoxicity, and chronic oxidative stress in type 2 diabetes. AB - The relentless decline in beta-cell function frequently observed in type 2 diabetic patients, despite optimal drug management, has variously been attributed to glucose toxicity and lipotoxicity. The former theory posits hyperglycemia, an outcome of the disease, as a secondary force that further damages beta-cells. The latter theory suggests that the often-associated defect of hyperlipidemia is a primary cause of beta-cell dysfunction. We review evidence that patients with type 2 diabetes continually undergo oxidative stress, that elevated glucose concentrations increase levels of reactive oxygen species in beta-cells, that islets have intrinsically low antioxidant enzyme defenses, that antioxidant drugs and overexpression of antioxidant enzymes protect beta-cells from glucose toxicity, and that lipotoxicity, to the extent it can be attributable to hyperlipidemia, occurs only in the context of preexisting hyperglycemia, whereas glucose toxicity can occur in the absence of hyperlipidemia. PMID- 14749277 TI - ALS/Lt: a new type 2 diabetes mouse model associated with low free radical scavenging potential. AB - Outbred CD-1 mice carry a spectrum of genetic susceptibilities for obesity and type 2 diabetes. ALS is an inbred strain with low antioxidant defenses produced by inbreeding CD-1 mice, with selection for susceptibility to alloxan, a generator of highly reactive oxygen free radicals and a potent beta-cell toxin. The objective of this study was to determine if the low ability to diffuse free radical stress would contribute to spontaneous type 2 diabetes development in alloxan-untreated males. Indeed, both hyperinsulinemia and impaired glucose tolerance developed spontaneously between 6 and 8 weeks of age in alloxan untreated males. Further aging was accompanied by increases in body mass, progressively more severe hyperinsulinemia, and development of overt hyperglycemia. Transition from impaired glucose tolerance to overt hyperglycemia correlated with a decreased ratio of reduced to oxidized glutathione. Evidence that the increased oxidative burden elicited the type 2 diabetes syndrome was obtained by the systemic elevation of the antioxidative capacity through daily administration of R-lipoic acid. R-lipoic acid (30 mg/kg) prevented hyperglycemia, reduced insulin levels, and increased free radical scavenging potential. This mouse model with reduced ability to diffuse free radical stress is of obvious interest because free radical-mediated damage is implicated in the pathogenesis and complications of both type 1 and type 2 diabetes. PMID- 14749278 TI - The biology of mitochondrial uncoupling proteins. AB - Uncoupling proteins (UCPs) are mitochondrial transporters present in the inner membrane of mitochondria. They are found in all mammals and in plants. They belong to the family of anion mitochondrial carriers including adenine nucleotide transporters. The term "uncoupling protein" was originally used for UCP1, which is uniquely present in mitochondria of brown adipocytes, the thermogenic cells that maintain body temperature in small rodents. In these cells, UCP1 acts as a proton carrier activated by free fatty acids and creates a shunt between complexes of the respiratory chain and ATP synthase. Activation of UCP1 enhances respiration, and the uncoupling process results in a futile cycle and dissipation of oxidation energy as heat. UCP2 is ubiquitous and highly expressed in the lymphoid system, macrophages, and pancreatic islets. UCP3 is mainly expressed in skeletal muscles. In comparison to the established uncoupling and thermogenic activities of UCP1, UCP2 and UCP3 appear to be involved in the limitation of free radical levels in cells rather than in physiological uncoupling and thermogenesis. Moreover, UCP2 is a regulator of insulin secretion and UCP3 is involved in fatty acid metabolism. PMID- 14749279 TI - Uncoupling protein 2 and islet function. AB - Stressors such as chronic hyperglycemia or hyperlipidemia may lead to insufficient insulin secretion in susceptible individuals, contributing to type 2 diabetes. The molecules mediating this effect are just beginning to be identified. Uncoupling protein (UCP)-2 may be one such negative modulator of insulin secretion. Accumulating evidence shows that beta-cell UCP2 expression is upregulated by glucolipotoxic conditions and that increased activity of UCP2 decreases insulin secretion. Mitochondrial superoxide has been identified as a posttranslational regulator of UCP2 activity in islets; thus, UCP2 may provide protection to beta-cells at one level while simultaneously having detrimental effects on insulin secretion. Interestingly, the latter appears to be the dominant outcome, because UCP2 knockout mice display an increased beta-cell mass and retained insulin secretion capacity in the face of glucolipotoxicity. PMID- 14749280 TI - Update on adipocyte hormones: regulation of energy balance and carbohydrate/lipid metabolism. AB - Hormones produced by adipose tissue play a critical role in the regulation of energy intake, energy expenditure, and lipid and carbohydrate metabolism. This review will address the biology, actions, and regulation of three adipocyte hormones-leptin, acylation stimulating protein (ASP), and adiponectin-with an emphasis on the most recent literature. The main biological role of leptin appears to be adaptation to reduced energy availability rather than prevention of obesity. In addition to the well-known consequences of absolute leptin deficiency, subjects with heterozygous leptin gene mutations have low circulating leptin levels and increased body adiposity. Leptin treatment dramatically improves metabolic abnormalities (insulin resistance and hyperlipidemia) in patients with relative leptin deficiency due to lipoatrophy. Leptin production is primarily regulated by insulin-induced changes of adipocyte metabolism. Dietary fat and fructose, which do not increase insulin secretion, lead to reduced leptin production, suggesting a mechanism for high-fat/high-sugar diets to increase energy intake and weight gain. ASP increases the efficiency of triacylglycerol synthesis in adipocytes leading to enhanced postprandial lipid clearance. In mice, ASP deficiency results in reduced body fat, obesity resistance, and improved insulin sensitivity. Adiponectin production is stimulated by thiazolidinedione agonists of peroxisome proliferator-activated receptor-gamma and may contribute to increased insulin sensitivity. Adiponectin and leptin cotreatment normalizes insulin action in lipoatrophic insulin-resistant animals. These effects may be mediated by AMP kinase-induced fat oxidation, leading to reduced intramyocellular and liver triglyceride content. The production of all three hormones is influenced by nutritional status. These hormones, the pathways controlling their production, and their receptors are promising targets for managing obesity, hyperlipidemia, and insulin resistance. PMID- 14749281 TI - Leptin effects on pancreatic beta-cell gene expression and function. AB - The hormone leptin is secreted from white adipocytes, and serum levels of leptin correlate with adipose tissue mass. Leptin was first described to act on the satiety center in the hypothalamus through specific receptors (leptin receptor [ObR]) to restrict food intake and enhance energy expenditure. Important peripheral actions of leptin involve inhibition of insulin biosynthesis and secretion in pancreatic beta-cells. In turn, insulin stimulates leptin secretion from adipose tissue, establishing a hormonal regulatory feedback loop-the so called "adipo-insular axis." Multiple signal transduction pathways are involved in leptin signaling in pancreatic beta-cells. We have identified the proinsulin gene and protein phosphatase 1 gene as leptin repressed genes and the gene for the suppressor of cytokine signaling 3 protein as a leptin-induced gene in pancreatic beta-cells. The molecular effects of leptin culminate to restrict insulin secretion and biosynthesis to adapt glucose homeostasis to the amount of body fat. In most overweight individuals, however, physiological regulation of body weight by leptin seems to be disturbed, representing "leptin resistance." This leptin resistance at the level of the pancreatic beta-cell may contribute to dysregulation of the adipo-insular axis and promote the development of hyperinsulinemia and manifest type 2 diabetes in overweight patients. PMID- 14749282 TI - Chronic effects of fatty acids on pancreatic beta-cell function: new insights from functional genomics. AB - Type 2 diabetes can be viewed as a failure of the pancreatic beta-cell to compensate for peripheral insulin resistance with enhanced insulin secretion. This failure is explained by both a relative loss of beta-cell mass as well as secretory defects that include enhanced basal secretion and a selective loss of sensitivity to glucose. These features are reproduced by chronic exposure of beta cells to fatty acids (FAs), suggesting that hyperlipidemia might contribute to decompensation. Using MIN6 cells pretreated for 48 h with oleate or palmitate, we have previously defined alterations in global gene expression by transcript profiling and described additional secretory changes to those already established (Busch A-K, Cordery D, Denyer G, Biden TJ: Diabetes 51:977-987, 2002). In contrast to a modest decoupling of glucose-stimulated insulin secretion, FA pretreatment markedly enhanced the secretory response to an acute subsequent challenge with FAs. We propose that this apparent switch in sensitivity from glucose to FAs would be an appropriate response to hyperlipidemia in vivo and thus plays a positive role in beta-cell compensation for insulin resistance. Altered expression of dozens of genes could contribute to this switch, and allelic variations in any of these genes could (to varying degrees) impair beta cell compensation and thus contribute to conditions ranging from impaired glucose tolerance to frank diabetes. PMID- 14749283 TI - Acute omega-3 fatty acid enrichment selectively reverses high-saturated fat feeding-induced insulin hypersecretion but does not improve peripheral insulin resistance. AB - In rats fed a high-saturated fat diet, replacement of a small percentage of total fatty acids with long-chain omega-3 fatty acids from fish oil for the duration of high-fat feeding prevents the development of insulin resistance. We investigated the effect of acute (24-h) modulation of dietary fat composition on glucose stimulated insulin secretion (GSIS) in rats made insulin resistant by high saturated fat feeding for 4 weeks. Insulin secretion after an intravenous glucose challenge was greatly increased by high-saturated fat feeding. Glucose tolerance was minimally perturbed, demonstrating insulin hypersecretion compensated for insulin resistance. The effect of high-saturated fat feeding to enhance GSIS was retained in perifused islets, such that glucose stimulus-secretion coupling was potentiated. Acute replacement of 7% of dietary fatty acids with long-chain omega 3 fatty acids reversed insulin hypersecretion in vivo, and the effect of long term high-saturated fat feeding to enhance insulin secretion by perifused islets was also completely reversed. Although a hyperbolic relationship existed between insulin secretion and action in the high-saturated fat and control groups, lowered insulin secretion in the acute fish oil-supplemented group was not accompanied by improved insulin action, and glucose tolerance was adversely affected. Our studies are important because they demonstrate that hyperinsulinemia can be rapidly reversed via the dietary provision of small amounts of long-chain omega-3 fatty acids. However, this "insulin sparing" action of acute dietary long-chain omega-3 fatty acids occurs in the absence of an acute improvement in insulin sensitivity and therefore at the expense of maintenance of glucose tolerance. PMID- 14749284 TI - The role of cytosolic phospholipase A(2) in insulin secretion. AB - Cytosolic phospholipase A(2) (cPLA(2)) comprises a widely expressed family of enzymes, some members of which have the properties required of signal transduction elements in electrically excitable cells. Thus, alpha- and beta isoforms of cPLA(2) are activated by the increases in intracellular Ca(2+) concentration ([Ca(2+)](i)) achieved in depolarized cells. Activation is associated with a redistribution of the enzyme within the cell; activation of cPLA(2) generates arachidonic acid (AA), a biologically active unsaturated fatty acid that can be further metabolized to generate a plethora of biologically active molecules. Studies using relatively nonselective pharmacological inhibitors have implicated cPLA(2) in insulin secretory responses to stimuli that elevate beta-cell [Ca(2+)](i); therefore, we have investigated the role of cPLA(2) in beta-cell function by generating beta-cell lines that under- or overexpress the alpha-isoform of cPLA(2). The functional phenotype of the modified cells was assessed by observation of cellular ultrastructure, by measuring insulin gene expression and insulin protein content, and by measuring the effects of insulin secretagogues on cPLA(2) distribution, on changes in [Ca(2+)](i), and on the rate and pattern of insulin secretion. Our results suggest that cPLA(2) is not required for the initiation of insulin secretion from beta-cells, but that it plays an important role in the maintenance of beta-cell insulin stores. Our data also demonstrate that excessive production of, or exposure to, AA is deleterious to normal beta-cell secretory function through metabolic dysfunction. PMID- 14749286 TI - Beta-cell calcium-independent group VIA phospholipase A(2) (iPLA(2)beta): tracking iPLA(2)beta movements in response to stimulation with insulin secretagogues in INS-1 cells. AB - Evidence that group VIA cytosolic calcium-independent phospholipase A(2) (iPLA(2)beta) participates in beta-cell signal transduction includes the observations that inhibition of iPLA(2)beta with the bromoenol lactone suicide substrate suppresses glucose-stimulated insulin secretion and that overexpression of iPLA(2)beta amplifies insulin secretory responses in INS-1 insulinoma cells. Immunofluorescence analyses also reveal that iPLA(2)beta accumulates in the perinuclear region of INS-1 cells stimulated with glucose and forskolin. To characterize this phenomenon further, iPLA(2)beta was expressed as a fusion protein with enhanced green fluorescent protein (EGFP) in INS-1 cells so that movements of iPLA(2)beta are reflected by changes in the subcellular distribution of green fluorescence. Stimulation of INS-1 cells overexpressing iPLA(2)beta-EGFP induced greater insulin secretion and punctate accumulation of iPLA(2)beta-EGFP fluorescence in the perinuclear region. To determine the identity of organelles with which iPLA(2)beta might associate, colocalization of green fluorescence with fluorophores associated with specific trackers targeted to different subcellular organelles was examined. Such analyses reveal association of iPLA(2)beta-EGFP fluorescence with the ER and Golgi compartments. Arachidonate-containing plasmenylethanolamine phospholipid species are abundant in beta-cell endoplasmic reticulum (ER) and are excellent substrates for iPLA(2)beta. Arachidonic acid produced by iPLA(2)beta-catalyzed hydrolysis of their substrates induces release of Ca(2+) from ER stores-an event thought to participate in glucose-stimulated insulin secretion. PMID- 14749285 TI - Islet complex lipids: involvement in the actions of group VIA calcium-independent phospholipase A(2) in beta-cells. AB - The beta-isoform of group VIA calcium-independent phospholipase A(2) (iPLA(2)beta) does not require calcium for activation, is stimulated by ATP, and is sensitive to inhibition by a bromoenol lactone suicide substrate. Several potential functions have been proposed for iPLA(2)beta. Our studies indicate that iPLA(2)beta is expressed in beta-cells and participates in glucose-stimulated insulin secretion but is not involved in membrane phospholipid remodeling. If iPLA(2)beta plays a signaling role in glucose-stimulated insulin secretion, then conditions that impair iPLA(2)beta functions might contribute to the diminished capacity of beta-cells to secrete insulin in response to glucose, which is a prominent characteristic of type 2 diabetes. Our recent studies suggest that iPLA(2)beta might also participate in beta-cell proliferation and apoptosis and that various phospholipid-derived mediators are involved in these processes. Detailed characterization of the iPLA(2)beta protein level reveals that beta cells express multiple isoforms of the enzyme, and our studies involve the hypothesis that different isoforms have different functions. PMID- 14749287 TI - Glucose-induced regulation of COX-2 expression in human islets of Langerhans. AB - Cyclo-oxygenase (COX), the enzyme responsible for conversion of arachidonic acid to prostanoids, exists as two isoforms. In most tissues, COX-1 is a constitutive enzyme involved in prostaglandin-mediated physiological processes, whereas COX-2 is thought to be induced by inflammatory stimuli. However, it has previously been reported that COX-2 is the dominant isoform in islets and an insulin-secreting beta-cell line under basal conditions. We have investigated the relative abundance of COX-1 and COX-2 mRNAs in MIN6 cells, a mouse insulin-secreting cell line, and in primary mouse and human islets. We found that COX-2 was the dominant isoform in MIN6 cells, but that COX-1 mRNA was more abundant than that of COX-2 in freshly isolated mouse islets. Furthermore, COX-2 expression was induced by maintenance of mouse islets in culture, and experiments with human islets indicated that exposure of the islets to hyperglycemic conditions was sufficient to upregulate COX-2 mRNA levels. Given that hyperglycemia has been reported to increase human beta-cell production of interleukin-1beta and that this cytokine can induce COX-2 expression, our observations of glucose-induced induction of COX 2 in human islets suggest that this is one route through which hyperglycemia may contribute to beta-cell dysfunction. PMID- 14749288 TI - Facial pain increases nausea and headache during motion sickness in migraine sufferers. AB - The aim of this study was to determine whether trigeminal nerve discharge associated with painful stimulation of the temple would intensify symptoms of motion sickness in migraine sufferers. If so, this would support the notion that symptoms such as nausea and headache interact with each other during attacks of migraine. Symptoms of motion sickness were rated at 2 min intervals during 15 min of optokinetic stimulation in 27 migraine sufferers and 23 age- and sex-matched controls. To document changes in frontotemporal blood flow, pulse amplitude was monitored with photoelectric pulse transducers. To induce facial pain, ice was applied to the temple for 30 s, three times at 4 min intervals during optokinetic stimulation. On another occasion, pain was induced during optokinetic stimulation by immersing the non-dominant hand in 2 degrees C ice water for 30 s, three times at 4 min intervals. On a third occasion, measures were obtained during optokinetic stimulation alone. Migraine sufferers rated themselves as being generally more susceptible to motion sickness than controls. In addition, symptoms of motion sickness provoked by optokinetic stimulation were greater in migraine sufferers than in controls. Painful stimulation of the temple intensified nausea and headache during optokinetic stimulation, whereas painful stimulation of the hand did not. Since nausea also intensifies facial pain during motion sickness, nausea and headache may reinforce each other in a vicious circle. In the absence of painful stimulation, increases in pulse amplitude during optokinetic stimulation were greater in migraine sufferers than controls, possibly because the discomfort associated with motion sickness triggered extracranial vasodilatation in migraine sufferers as part of a fight-or-flight (defense) response. Extracranial vasodilatation did not differ between migraine sufferers and controls when ice was applied to the temple or hand during optokinetic stimulation, implying that the additional discomfort associated with painful stimulation of the head and hand evoked a defense response in controls. These findings suggest that a mechanism which boosts extracranial neurovascular reflexes to stress and which heightens symptoms of motion sickness, increases susceptibility to migraine. PMID- 14749289 TI - fMRI correlates of state and trait effects in subjects at genetically enhanced risk of schizophrenia. AB - Schizophrenia is a highly heritable disorder that typically develops in early adult life. Structural imaging studies have indicated that patients with the illness, and to some extent their unaffected relatives, have subtle deficits in several brain regions, including prefrontal and temporal lobes. It is, however, not known how this inherited vulnerability leads to psychosis. This study used a covert verbal initiation fMRI task previously shown to elicit frontal and temporal activity (the Hayling sentence completion task) to examine this issue. A large (n = 69) number of young participants at high risk of developing schizophrenia for genetic reasons took part, together with a matched group of healthy controls (n = 21). At the time of investigation, none had any psychotic disorder, but on detailed interview some of the high-risk participants (n = 27) reported isolated psychotic symptoms. The study aimed to determine: (i) whether there were activation differences that occurred in all subjects with a genetic risk of schizophrenia (i.e. 'trait' effects); and (ii) whether there were activation differences that only occurred in those at high risk who had isolated psychotic symptoms ('state' effects). No activation differences were found in regions commonly reported to be abnormal in the established illness, namely the dorsolateral prefrontal cortex or in the temporal lobes, but group differences of apparent genetic cause were evident in medial prefrontal, thalamic and cerebellar regions. In addition, differences in activation in those with symptoms were found in the intraparietal sulcus. No significant differences in performance were found between the groups, and all subjects were antipsychotic naive. These findings therefore suggest that vulnerability to schizophrenia may be inherited as a disruption in a fronto-thalamic-cerebellar network, and the earliest changes specific to the psychotic state may be related to hyperactivation in the parietal lobe. PMID- 14749290 TI - Forces consistent with plateau-like behaviour of spinal neurons evoked in patients with spinal cord injuries. AB - Percutaneous electrical stimulation over tibialis anterior and triceps surae was performed in 14 patients with traumatic spinal cord injury (SCI) to look for evidence that 'extra contractions' can develop, beyond those due to activation of the motor axons beneath the stimulating electrodes. Criteria for the extra contractions included marked asymmetry of force with respect to stimulation, progressively rising force during stimulation of constant amplitude and frequency, and force remaining high after stimulation frequency had returned to the control level following a high-frequency burst. Twelve of the 14 patients showed evidence of such behaviour, more frequently in triceps surae than tibialis anterior. Force or electromyographic activity commonly outlasted the stimulation in these patients. There was no apparent correlation between the completeness or level of injury and the ability to induce the behaviour. Evidence of force potentiation and 'habituation' was also seen. Eleven of the 14 patients exhibited hyper-reflexia and reported spontaneous spasms, but there was no obvious association with the extra contractions. It is concluded that non-classical behaviour of neurons within the spinal cord can contribute to the extra contractions evoked by electrical stimulation over muscles in spinal cord-injured subjects. This central contribution is less easy to obtain than in intact healthy subjects, all of whom showed the phenomenon. These contractions are consistent with the activation of plateau potentials in spinal neurons and, if so, plateau potentials may contribute to a patient's clinical manifestations. PMID- 14749291 TI - Reorganization of the human ipsilesional premotor cortex after stroke. AB - The substrates that mediate recovery of motor function after stroke are incompletely understood. Several primate and human studies proposed the involvement of the premotor cortex of the lesioned hemisphere. Here, we studied four chronic stroke patients with focal subcortical lesions affecting the corticospinal outflow originating in the primary motor cortex (M1) and good motor recovery. We tested the hypothesis that, in these patients, disruption of activity in the premotor cortex of the lesioned hemisphere by transcranial magnetic stimulation (TMS) would result in degraded behaviour in the paretic hand. TMS was applied to the primary motor cortex, dorsal premotor cortex (PMd) and ventral premotor cortex (PMv) of the affected (M1AH, PMdAH, PMvAH) and intact (M1IH, PMdIH, PMvIH) hemispheres of patients and healthy controls in the setting of a simple reaction time (SRT) paradigm performed with the hand contralateral to the stimulated hemisphere. TMS applied to M1 led to substantial contralateral SRT delays in both groups. TMS applied to PMdAH of patients elicited clear delays in contralateral SRT in the paretic hand, whereas TMS applied to PMdIH of patients or healthy volunteers did not. Motor evoked potentials after stimulation of PMdAH were, on average, larger and had, on average, shorter latency than after stimulation of M1AH. These results indicate that PMdAH participates as a substrate mediating functional recovery of executive motor function in patients with focal lesions of corticospinal outflow originating in M1 and good motor recovery. Our results are consistent with the hypothesis that the dorsal premotor cortex of the affected hemisphere can reorganize to control basic parameters of movement usually assigned to M1 function. PMID- 14749292 TI - Cerebral atrophy in Parkinson's disease with and without dementia: a comparison with Alzheimer's disease, dementia with Lewy bodies and controls. AB - Parkinson's disease is a common neurodegenerative disorder primarily characterized by rigidity, tremor and bradykinesia. Cognitive impairment and neuropsychiatric symptoms are frequent in Parkinson's disease, with a 70% cumulative incidence of dementia. The aim of this cross-sectional study was to establish the pattern of cerebral atrophy on MRI in Parkinson's disease patients with dementia. We used voxel-based morphometry (VBM) to provide an unbiased means of investigating brain volume loss. Whole brain structural T1-weighted MRI scans from Parkinson's disease patients with dementia (PDD, n = 26), Parkinson's disease patients without dementia (n = 31), Alzheimer's disease patients (n = 28), patients with dementia with Lewy bodies (DLB, n = 17) and control subjects (n = 36) were acquired. Images were analysed using SPM99 and the optimized method of VBM. Reduced grey matter volume in PDD patients compared with controls was observed bilaterally in the temporal lobe, including the hippocampus and parahippocampal gyrus, and in the occipital lobe, the right frontal lobe and the left parietal lobe, as well as some subcortical regions. Parkinson's disease patients without dementia showed reduced grey matter volume in the frontal lobe compared with control subjects. There was significant grey matter atrophy bilaterally in the occipital lobe of PDD patients compared with Parkinson's disease patients. In addition, significant temporal lobe atrophy, including the hippocampus and parahippocampal gyrus was detected in Alzheimer's disease relative to PDD. No significant volumetric differences were observed in PDD compared with DLB. Thus, Parkinson's disease involves grey matter loss in frontal areas. In PDD, this extends to temporal, occipital and subcortical areas, with occipital atrophy in PDD being the only difference between the two groups. This provides important information about the pattern of cerebral atrophy in Parkinson's disease and PDD. PMID- 14749293 TI - Protective effect of herpes simplex virus-mediated neurotrophin gene transfer in cisplatin neuropathy. AB - Attempts to develop clinical treatments for neuropathy using neurotrophins have not been successful. We tested whether neurotrophin gene delivery to dorsal root ganglia (DRGs) using non-replicating herpes simplex virus (HSV)-based vectors could prevent the development of neuropathy caused by administration of cisplatin. Following subcutaneous inoculation of HSV vectors expressing nerve growth factor (NGF) or neurotrophin-3 (NT-3), neurons in the DRG were transduced to produce NGF or NT-3 in vivo. Inoculation of either the NGF- or the NT-3 expressing vectors 3 days before the start of a 6-week course of cisplatin treatment protected against cisplatin-induced neuropathy assessed by electrophysiological, histological and behavioural measures 2 months later. Iatrogenic neuropathy caused by administration of chemotherapeutic drugs represents an excellent target for a human trial to assess the potential of gene therapy to prevent neuropathy. PMID- 14749294 TI - Verbal recall in amnesiacs under conditions of diminished retroactive interference. AB - In amnesiacs, stimuli that at first can be recalled are usually forgotten within 1 min, but the conditions required for this severe forgetting have remained unknown. To examine this, six patients with amnesia due to head injury or stroke and six normal controls heard lists of words (Experiment 1) and stories (Experiment 2). These stimuli were to be recalled immediately or after an extended test delay (10 min in Experiment 1; 1 h in Experiment 2). Although severe forgetting occurred in the amnesiacs following activity-filled delays, much less forgetting occurred in four of these patients after delays spent in a dark, quiet room. This was true even when the patients appeared to sleep during the delays. The results show, in a novel manner, that one deficit underlying their amnesias is vulnerability to retroactive interference. PMID- 14749295 TI - Regulation of gonadotropin-releasing hormone (GnRH)-receptor gene expression in tilapia: effect of GnRH and dopamine. AB - The present work was designed to study certain aspects of the endocrine regulation of gonadotropin-releasing hormone receptor (GnRH-R) in the pituitary of the teleost fish tilapia. A GnRH-R was cloned from the pituitary of hybrid tilapia (taGnRH-R) and was identified as a typical seven-transmembrane receptor. Northern blot analysis revealed a single GnRH-R transcript in the pituitary of approximately 2.3 kilobases. The taGnRH-R mRNA levels were significantly higher in females than in males. Injection of the salmon GnRH analog (sGnRHa; 5-50 microg/kg) increased the steady-state levels of taGnRH-R mRNA, with the highest response recorded at 25 microg/kg and at 36 h. At the higher dose of sGnRHa (50 microg/kg), taGnRH-R transcript appeared to be down-regulated. Exposure of tilapia pituitary cells in culture to graded doses (0.1-100 nM) of seabream (sbGnRH = GnRH I), chicken II (cGnRH II), or salmon GnRH (sGnRH = GnRH III) resulted in a significant increase in taGnRH-R mRNA levels. The highest levels of both LH release and taGnRH-R mRNA levels were recorded after exposure to cGnRH II and the lowest after exposure to sbGnRH. The dopamine-agonist quinpirole suppressed LH release and mRNA levels of taGnRH-R, indicating an inhibitory effect on GnRH-R synthesis. Collectively, these data provide evidence that GnRH in tilapia can up- regulate, whereas dopamine down-regulates, taGnRH-R mRNA levels. PMID- 14749296 TI - Telomere lengths in cloned transgenic pigs. AB - Studies of cloned cattle and mice have resulted in controversies regarding the restoration of eroded telomere length of donor cells by the nuclear transfer process. Little is known about telomere lengths in pigs from either natural reproduction or nuclear transfer. In this study, we measured the telomere lengths in six major porcine organs from animals of different ages, and found that their lengths remained consistent throughout different tissues during fetal stages, and then shortened, in a tissue- specific manner, after birth. Telomeres of skin samples from six cloned transgenic pigs at 4 mo of age did not differ significantly from those of age-matched controls. Two cloned pigs that died shortly after birth had skin telomere lengths equivalent to those of late-stage fetuses. PMID- 14749297 TI - A morphological and immunohistochemical comparison of mammary tissues from the short-tailed fruit bat (Carollia perspicillata) and the mouse. AB - In the present study, mammary tissues from the fruit bat (Carollia perspicillata) and mouse (Mus musculus) were compared using histological and immunohistochemical methods. Because the female bat exhibits greater reproductive similarities to humans, it might provide a useful animal model for studying mammary physiology and disease with relevance to our own species. In lactating and recently lactating specimens, bat tissue had significantly fewer adipocytes and more collagenous connective tissue compared to the mouse. The proteins Stat5a, keratin 5, Npt2b, and E-cadherin were all similarly localized in mouse and bat mammary tissues taken from lactating animals. The present study demonstrates that whereas the epithelial compartment and the presence of differentiation markers are conserved between the mouse and bat, differences exist in the stromal compartment. PMID- 14749298 TI - Secretin controls anion secretion in the rat epididymis in an autocrine/paracrine fashion. AB - There is growing evidence that secretin, the first hormone discovered in our history, has functions in the brain other than in the gastrointestinal tract. This article reports for the first time that secretin and its receptor mRNAs are produced in distinct cell types within the epididymis. To test if secretin affects electrolyte transport in the epididymis, we measured short-circuit current (Isc) in cultured epididymal epithelia and found secretin dose dependently stimulated Isc. Ion substitution experiments and use of pharmacological agents inferred that the stimulated Isc is a result of concurrent electrogenic chloride and bicarbonate secretion. It is further shown that secretin and pituitary adenylate cyclase-activating polypeptide (PACAP) function via totally different mechanisms: 1) PACAP works only from the apical side of the epithelium to stimulate chloride and not bicarbonate secretion, while secretin acts on the apical and basolateral sides to stimulate chloride and bicarbonate secretion. 2) the stimulation by PACAP but not secretin requires local prostaglandin synthesis. By immunocytochemical staining, secretin is localized in the principal cells of the initial segment and caput epididymidis, whereas secretin receptor is present in the principal cells of the proximal as well as the distal part of the epididymis. This pattern of distribution appears to be consistent with the idea that secretin is secreted by the proximal epididymis and acts on the proximal and distal epididymis in an autocrine and paracrine fashion. Its function is to control secretion of electrolytes and water. PMID- 14749299 TI - Mitochondria-dependent pathway is involved in heat-induced male germ cell death: lessons from mutant mice. AB - The signaling events leading to apoptosis can be divided into two major pathways, involving either mitochondria (intrinsic) or death receptors (extrinsic). In a recent study, we have shown the involvement of the mitochondria-dependent apoptotic pathway in heat-induced male germ cell apoptosis in the rat. In additional studies, using the gld (generalized lymphoproliferation disease) and lprcg (lymphoproliferation complementing gld) mice, which harbor loss-of-function mutations in Fas L and Fas, respectively, we have shown that heat-induced germ cell apoptosis is not blocked, thus providing evidence that the Fas signaling system is not required for heat-induced germ cell apoptosis in the testis. In the present study, we have found that the initiation of apoptosis in wild-type mice was preceded by a redistribution of Bax from a cytoplasmic to paranuclear localization in heat-susceptible germ cells. The relocation of Bax is accompanied by sequestration of ultracondensed mitochondria into paranuclear areas of apoptotic germ cells, cytosolic translocation of mitochondrial cytochrome c and DIABLO, and is associated with activation of the initiator caspase 9 and the executioner caspase 3. Similar events were also noted in both gld and lprcg mice. Taken together, these results indicate that the mitochondria-dependent pathway is the key apoptotic pathway for heat-induced male germ cell death in mice. PMID- 14749300 TI - Cell-specific expression and regulation of soluble guanylyl cyclase alpha 1 and beta 1 subunits in the rat ovary. AB - Soluble guanylyl cyclase (sGC) is activated by nitric oxide (NO) and carbon monoxide, resulting in cGMP production. Recent studies indicate that NO and cGMP influence ovarian functions. However, little information is available regarding the ovarian expression of sGC. The present study examined sGC alpha(1) and beta(1) subunit protein levels in the ovary during postnatal development, gonadotropin-induced follicle growth, ovulation, and luteinization as well as in cultured rat granulosa cells. In postnatal rats, sGC alpha(1) subunit immunoreactivity was high in granulosa cells of primordial and primary follicles on Day 5 but low in granulosa cells of larger follicles on Days 10 and 19. Theca cells of developing follicles, but not stromal cells, also demonstrated moderate sGC alpha(1) immunoreactivity. In gonadotropin- treated immature rats, intense sGC alpha(1) subunit staining was similarly observed in granulosa cells of primordial and primary follicles, but such staining was low in granulosa cells of small antral follicles and undetectable in granulosa cells of large antral and preovulatory follicles. Following ovulation, corpora lutea expressed moderate sGC alpha(1) immunoreactivity. Similar ovarian localization and expression patterns were seen for sGC beta(1), indicating regulated coexpression of sGC subunits. Immunoblot analysis revealed no change in total ovarian sGC alpha(1) and beta(1) subunit protein levels during gonadotropin treatment. Similarly, no effect of FSH on sGC subunit protein levels was apparent in cultured granulosa cells. These findings indicate regulated, cell- specific patterns of sGC expression in the ovary and are consistent with roles for cGMP in modulating ovarian functions. PMID- 14749301 TI - Abnormal morphology of the penis in male rats exposed neonatally to diethylstilbestrol is associated with altered profile of estrogen receptor-alpha protein, but not of androgen receptor protein: a developmental and immunocytochemical study. AB - Objectives of the study were to determine developmental changes in morphology and expression of androgen receptor (AR) and estrogen receptor (ER)alpha in the body of the rat penis exposed neonatally to diethylstilbestrol (DES). Male pups received DES at a dose of 10 microg per rat on alternate days from Postnatal Day 2 to Postnatal Day 12. Controls received olive oil vehicle only. Tissue samples were collected on Days 18 (prepuberty), 41 (puberty), and 120 (adult) of age. DES induced abnormalities were evident at 18 days of age and included smaller, lighter, and thinner penis, loss of cavernous spaces and associated smooth muscle cells, and increased deposition of fat cells in the corpora cavernosa penis. Fat cells virtually filled the entire area of the corpora cavernosa at puberty and adulthood. Plasma testosterone (T) was reduced to an undetectable level, while LH was unaltered in all treated groups. AR-positive cells were ubiquitous and their profile (incidence and staining intensity) did not differ between control and treated rats of the respective age groups. Conversely, ERalpha-positive cells were limited to the stroma of corpus spongiosus in all age groups of both control and treated rats, but the expression in treated rats at 18 days was up-regulated in stromal cells of corpora cavernosa, coincident with the presence of morphological abnormalities. Hence, this study reports for the first time DES induced developmental, morphological abnormalities in the body of the penis and suggests that these abnormalities may have resulted from decreased T and/or overexpression of ERalpha. PMID- 14749302 TI - Downregulation of follicle-stimulating hormone (FSH)-receptor messenger RNA levels in the hamster ovary: effect of the endogenous and exogenous FSH. AB - Although gonadotropins have been reported to downregulate FSH-receptor (FSHR) mRNA levels in the ovaries of female rats, the effect of the gonadotropin surge, particularly FSH, on hamster follicular FSHR mRNA levels warrants further examination. The objectives of the present study were to clone and determine the complete FSHR cDNA sequence of the hamster and to delineate the effects of endogenous and exogenous FSH on the steady-state levels of ovarian FSHR mRNA. Complete FSHR cDNA was derived from hamster ovarian total RNA by the strategy of 3'- and 5'-rapid amplification of cDNA ends. Ovaries were obtained before and after the endogenous gonadotropin surge or exogenous FSH administration, and the steady-state levels of FSHR mRNA were assessed by Northern blot hybridization. Cloned FSHR cDNA consists of a reading frame corresponding to exons 1-10 of the human FSHR gene and the 5'- and 3'-untranslated regions. The nucleic acid and amino acid sequences of the reading frame were at least 87% and 92% identical, respectively, to that of human, rat, and mouse FSHR. Furthermore, the amino acid sequence contained seven transmembrane domains characteristic of the FSHR. The steady-state levels of FSHR mRNA increased from estrus (Day 1) to reach a peak on proestrus (Day 4) noon; however, significant attenuation was noted following the gonadotropin surge, which was blocked by phenobarbital. Exogenous FSH also downregulated, both dose- and time-dependently, ovarian FSHR mRNA levels. These data indicate that the nucleic acid sequence of hamster FSHR has been identified and that FSH modulates FSHR mRNA levels in the hamster ovary. PMID- 14749303 TI - Primary afferent stimulation differentially potentiates excitatory and inhibitory inputs to spinal lamina II outer and inner neurons. AB - Spinal lamina II (substantia gelatinosa) neurons play an important role in processing of nociceptive information from primary afferent nerves. Anatomical studies suggest that neurons in the outer (lamina II(o)) and inner (lamina II(i)) zone of lamina II receive distinct afferent inputs. The functional significance of this preferential afferent termination in lamina II remains unclear. In this study, we examined the differential synaptic inputs to neurons in lamina II(o) and II(i) in response to primary afferent stimulation. Whole cell voltage-clamp recordings were performed on neurons in lamina II(o) and II(i) of the rat spinal cord slice under visual guidance. Capsaicin (1 microM) significantly increased the frequency of glutamatergic miniature excitatory postsynaptic currents (mEPSCs) in all 27 lamina II(o) neurons and significantly increased the amplitude of mEPSCs in 12 of 27 lamina II(o) neurons. However, capsaicin only significantly increased the frequency of mEPSCs in 9 of 22 (40.9%) lamina II(i) neurons and increased the amplitude of mEPSCs in 6 of these 9 neurons. Furthermore, the peak amplitude of EPSCs, evoked by electrical stimulation of the attached dorsal root, in 40 lamina II(o) neurons was significantly greater than that [160.5 +/- 16.7 vs. 87.0 +/- 10.4 (SE) pA] in 37 lamina II(i) neurons. On the other hand, the peak amplitude of evoked inhibitory postsynaptic currents (IPSCs) in 40 lamina II(o) neurons was significantly smaller than that (103.1 +/- 11.6 vs. 258.4 +/- 24.4 pA) in 37 lamina II(i) neurons. In addition, the peak amplitudes of both EPSCs and IPSCs, evoked by direct stimulation of lamina II, were similar in lamina II(o) and II(i) neurons. This study provides new information that stimulation of primary afferents differentially potentiates synaptic inputs to neurons in lamina II(o) and II(i). The quantitative difference in excitatory and inhibitory synaptic inputs to lamina II(o) and II(i) neurons may be important for integration of sensory information from primary afferent nerves. PMID- 14749304 TI - Neuronal encoding of ultrasonic sound by a fish. AB - Many species of odontocete cetaceans (toothed whales) use high-frequency clicks (60-170 kHz) to identify objects in their environment, including potential prey. Behavioral studies have shown that American shad, Alosa sapidissima, can detect ultrasonic signals similar to those of odontocetes that are potentially their predators. American shad also show strong escape behavior in response to ultrasonic pulses between 70 and 110 kHz and can determine the location of the sound source at least in the horizontal plane. The present study examines physiological aspects of ultrasound detection by American shad and provides the first insights into the neural encoding of ultrasound signals in any nonmammalian vertebrate. The recordings were obtained by penetration through the cerebellar surface. All but two units responded exclusively to ultrasound. Ultrasound sensitive units did not phase-couple to any stimulus frequency. Some units resembled the response of constant latency neurons found in the ventral nucleus of the lateral lemniscus of bats. We suggest that ultrasonic and sonic signals are processed along different pathways in Alosa. The ultrasonic pathway in Alosa appears to be a feature detector that is likely to be adapted (e.g., frequency, intensity) to odontocete echolocation signals. PMID- 14749305 TI - Interactions between light-induced currents, voltage-gated currents, and input signal properties in Drosophila photoreceptors. AB - Voltage-gated K(+) channels are important in neuronal signaling, but little is known of their interactions with receptor currents or their behavior during natural stimulation. We used nonparametric and parametric nonlinear modeling of experimental responses, combined with Hodgkin-Huxley style simulation, to examine the roles of K(+) channels in forming the responses of wild-type (WT) and Shaker mutant (Sh(14)) Drosophila photoreceptors to naturalistic stimulus sequences. Naturalistic stimuli gave results different from those of similar experiments with white noise stimuli. Sh(14) responses were larger and faster than WT. Simulation indicated that, in addition to eliminating the Shaker current, the mutation changed the current flowing through light-dependent channels [light induced current (LIC)] and increased the delayed rectifier current. Part of the change in LIC could be attributed to direct feedback from the voltage-sensitive ion channels to the light-sensitive channels by the membrane potential. However, we argue that other changes occur in the light detecting machinery of Sh(14) mutants, possibly during photoreceptor development. PMID- 14749306 TI - Large releasable pool of synaptic vesicles in chick cochlear hair cells. AB - Hearing requires the hair cell synapse to maintain notable temporal fidelity (< or =1 ms) while sustaining neurotransmitter release for prolonged periods of time (minutes). Here we probed the properties and possible anatomical substrate of prolonged neurotransmitter release by using electrical measures of cell surface area as a proxy for neurotransmitter release to study hair cell exocytosis evoked by repetitive stimuli. We observed marked depression of exocytosis by chick tall hair cells. This exocytic depression cannot be explained by calcium current inactivation, presynaptic autoinhibition by metabotropic glutamate receptors, or postsynaptic receptor desensitization. Rather, cochlear hair cell exocytic depression resulted from the exhaustion of a functional vesicle pool. This releasable vesicle pool is large, totaling approximately 8,000 vesicles, and is nearly 10 times greater than the number of vesicles tethered to synaptic ribbons. Such a large functional pool suggests the recruitment of cytoplasmic vesicles to sustain exocytosis, important for maintaining prolonged, high rates of neural activity needed to encode sound. PMID- 14749307 TI - Intracortical pathways determine breadth of subthreshold frequency receptive fields in primary auditory cortex. AB - To examine the basis of frequency receptive fields in auditory cortex (ACx), we have recorded intracellular (whole cell) and extracellular (local field potential, LFP) responses to tones in anesthetized rats. Frequency receptive fields derived from excitatory postsynaptic potentials (EPSPs) and LFPs from the same location resembled each other in terms of characteristic frequency (CF) and breadth of tuning, suggesting that LFPs reflect local synaptic (including subthreshold) activity. Subthreshold EPSP and LFP receptive fields were remarkably broad, often spanning five octaves (the maximum tested) at moderate intensities (40-50 dB above threshold). To identify receptive-field features that are generated intracortically, we microinjected the GABA(A) receptor agonist muscimol (0.2-5.1 mM, 1-5 microl) into ACx. Muscimol dramatically reduced LFP amplitude and reduced receptive-field bandwidth, implicating intracortical contributions to these features but had lesser effects on CF response threshold or onset latency, suggesting minimal loss of thalamocortical input. Reversal of muscimol's inhibition preferentially at the recording site by diffusion from the recording pipette of the GABA(A) receptor antagonist picrotoxin (0.01-100 microM) disinhibited responses to CF stimuli more than responses to spectrally distant, non-CF stimuli. We propose that thalamocortical and intracortical pathways preferentially contribute to responses evoked by CF and non-CF stimuli, respectively, and that intracortical projections linking frequency representations determine the breadth of receptive fields in primary ACx. Broad, subthreshold receptive fields may distinguish ACx from subcortical auditory relay nuclei, promote integrated responses to spectrotemporally complex stimuli, and provide a substrate for plasticity of cortical receptive fields and maps. PMID- 14749308 TI - Effect of a central fixation light on auditory spatial responses in area LIP. AB - A recent report demonstrated that, while fixating a central light, lateral intraparietal area (LIP) neurons are not modulated by the location of auditory stimuli until monkeys learn to saccade to the location of an auditory stimulus. This finding suggests that auditory spatial responses in area LIP are dependent on auditory-saccadic training. We found that, in monkeys that had not been trained to make behavioral responses to auditory stimuli, LIP neurons are modulated by auditory-stimulus location when a central light is not present in the environment. These results indicate that LIP auditory responses are not wholly dependent on behavioral training with auditory stimuli. PMID- 14749309 TI - Level of arousal during the small irregular activity state in the rat hippocampal EEG. AB - The sleeping rat cycles between two well-characterized hippocampal physiological states, large irregular activity (LIA) during slow-wave sleep (SWS) and theta activity during rapid-eye-movement sleep (REM). A third, less well-characterized electroencephalographic (EEG) state, termed "small irregular activity" (SIA), has been reported to occur when an animal is startled out of sleep without moving and during active waking when it abruptly freezes. We recently found that the hippocampal population activity of a spontaneous sleep state whose EEG resembles SIA reflects the rat's current location in space, suggesting that it is also a state of heightened arousal. To test whether this spontaneous SIA state corresponds to the SIA state reported in the literature and to compare the level of arousal during SIA to the other well-characterized physiological states, we recorded unit activity from ensembles of hippocampal CA1 pyramidal cells, EEG from the hippocampus and the neocortex, and electromyography (EMG) from the dorsal neck musculature in rats presented with auditory stimuli while foraging for randomly scattered food pellets and while sleeping. Auditory stimuli presented during sleep reliably induced SIA episodes very similar to spontaneous SIA in hippocampal and neocortical EEG amplitudes and power spectra, EMG amplitude, and CA1 population activity. Both spontaneous and elicited SIA exhibited neocortical desynchronization, and both had EMG amplitude comparable to that of waking LIA. We conclude based on this and other evidence that spontaneous SIA and elicited SIA correspond to a single state and that the level of arousal in SIA is higher than in the well-characterized sleep states but lower than the active theta state. PMID- 14749310 TI - Cone inputs in macaque primary visual cortex. AB - To understand the role of primary visual cortex (V1) in color vision, we measured directly the input from the 3 cone types in macaque V1 neurons. Cells were classified as luminance-preferring, color-luminance, or color-preferring from the ratio of the peak amplitudes of spatial frequency responses to red/green equiluminant and to black/white (luminance) grating patterns, respectively. In this study we used L-, M-, and S-cone-isolating gratings to measure spatial frequency response functions for each cone type separately. From peak responses to cone-isolating stimuli we estimated relative cone weights and whether cone inputs were the same or opposite sign. For most V1 cells the relative S-cone weight was <0.1. All color-preferring cells were cone opponent and their L/M cone weight ratio was clustered around a value of -1, which is roughly equal and opposite L and M cone signals. Almost all cells (88%) classified as luminance cells were cone nonopponent, with a broad distribution of cone weights. Most cells (73%) classified as color-luminance cells were cone opponent. This result supports our conclusion that V1 color-luminance cells are double-opponent. Such neurons are more sensitive to color boundaries than to areas of color and thereby could play an important role in color perception. The color-luminance population had a broad distribution of L/M cone weight ratios, implying a broad distribution of preferred colors for the double-opponent cells. PMID- 14749311 TI - Gaze-related response properties of DLPN and NRTP neurons in the rhesus macaque. AB - The dorsolateral pontine nucleus (DLPN) and nucleus reticularis tegmenti pontis (NRTP) are basilar pontine nuclei important for control of eye movements. The aim of this study was to compare the response properties of neurons in DLPN and rostral NRTP (rNRTP) during visual, oculomotor, and vestibular testing. We tested 51 DLPN neurons that were modulated during smooth pursuit (23/51) or during motion of a large-field visual stimulus (28/51). Following vestibular testing, we found that the majority of smooth pursuit-related neurons in DLPN were best classified as gaze (13/23) or eye velocity (7/23) related. Only a small percentage (3/51) of DLPN neurons responded during vestibular ocular reflex in the dark (VORd). We tested rNRTP neurons as described above and found the majority of neurons (35/43) were modulated during smooth pursuit or during motion of a large-field stimulus only (4/43). A significant proportion of our rNRTP gaze velocity neurons (10/18) were also modulated during VORd. We found that the majority of smooth pursuit related neurons in rNRTP were best classified as gaze velocity (18/35) or gaze acceleration (11/35) sensitive. The remaining neurons were classified as eye position or eye/head related. We used multiple linear regression modeling to determine the relative contributions of eye, head and visual inputs to the responses of DLPN and rNRTP neurons. Our results support the suggestion that both DLPN and rNRTP play significant roles not only in control of smooth pursuit but also in control of gaze. PMID- 14749312 TI - Ethanol alters the frequency, amplitude, and decay kinetics of Sr2+-supported, asynchronous NMDAR mEPSCs in rat hippocampal slices. AB - To discriminate between pre- and postsynaptic effects of ethanol on N-methyl-d aspartate receptor (NMDAR) signaling in hippocampus, we adapted the technique of Sr(2+) substitution to the hippocampal blind slice patch-clamp preparation. Hippocampal slices were isolated from 12- to 20-day-old rats that were killed in accordance with University of Texas Institutional Animal Care and Use Committee guidelines. NMDAR miniature excitatory postsynaptic currents (mEPSCs) were evoked from CA1 pyramidal neurons in the presence of Sr(2+) (4 mM), causing the synchronous EPSC observed in the presence of Ca(2+) to be supplanted by asynchronous mEPSCs. Amplitudes typically ranged from 5 to 40 pA and responded to the NMDAR antagonist (DL)-APV (50 microM), with a statistically significant reduction in mean amplitude. Ethanol (25, 50, and 75 mM) exerted dose-dependent effects on mEPSC amplitude and frequency. Peak amplitude inhibition was observed at 75 mM ethanol. Notably, ethanol significantly decreased event frequency at 50 and 75 mM ethanol. Ethanol (75 mM) also significantly increased the paired-pulse ratio of NMDAR EPSCs. Cumulative comparisons of decay time constants derived from single-exponential fitting of mEPSCs revealed significantly accelerated current decay kinetics in the presence of 75 mM ethanol. Taken together, these reductions in miniature event frequency and amplitude, concurrent with an increased rate of decay, suggest that the acute effects of ethanol on NMDAR signaling at hippocampal synapses are multifocal in nature. This finding of pre- and postsynaptic effects of ethanol on NMDAR signal strength in a brain region central to cognition is wholly consistent with previous reports of ethanol inhibition of NMDAR-long-term potentiation in vitro and with the profound cognitive deficits associated with binge-level intoxication in vivo. PMID- 14749313 TI - Area-selective neuronal activity in the dorsolateral prefrontal cortex for information retrieval and action planning. AB - We compared how neurons in the dorsal and ventral regions of the dorsolateral prefrontal cortex (dl-PFC) participate in processing 2 sets of sensory signals, given at intervals, to generate plans for future actions. For the first set of visual signals, neurons in the ventral region of dl-PFC responded preferentially to the visuospatial properties of the signal, whereas neurons in the dorsal region of dl-PFC were involved primarily in retrieving information from the signal, such as the location of the target or which arm to use. For the second set of visual signals, most ventral dl-PFC neurons reflected either the sensory properties of the signals or the information retrieved from each signal. By contrast, dorsal neurons were involved more in integrating information about the target location and which arm to use to reach the target, thereby generating information that could be used to plan future actions. Thus sensorimotor transformations in the dorsolateral PFC appear to be time-variant and region selective. PMID- 14749314 TI - Cortical involvement in the recruitment of wrist muscles. AB - In executing a voluntary movement, one is faced with the problem of translating a specification of the movement in task space (e.g., a visual goal) into a muscle recruitment pattern. Among many brain regions, the primary motor cortex (MI) plays a prominent role in the specification of movements. In what coordinate frame MI represents movement has been a topic of considerable debate. In a two dimensional wrist step-tracking experiment, Kakei et al. described some MI cells as encoding movement in a muscle-coordinate frame and other cells as encoding movement in an extrinsic-coordinate frame. This result was interpreted as evidence for a cascade of transformations within MI from an extrinsic representation of movement to a muscle-like representation. However, we present a model that demonstrates that, given a realistic extrinsic-like representation of movement, a simple linear network is capable of representing the transformation from an extrinsic space to the muscle-recruitment patterns implementing the movements on which Kakei et al. focused. This suggests that cells exhibiting extrinsic-like qualities can be involved in the direct recruitment of spinal motor neurons. These results call into question models that presume a serial cascade of transformations terminating with MI pyramidal tract neurons that vary their activation exclusively with muscle activity. Further analysis of the model shows that the correlation between the activity of an MI neuron and a muscle does not predict the strength of the connection between the MI neuron and muscle. This result cautions against the use of correlation methods as a measure of cellular connectivity. PMID- 14749315 TI - Effects of search efficiency on surround suppression during visual selection in frontal eye field. AB - Previous research has shown that visually responsive neurons in the frontal eye field of macaque monkeys select the target for a saccade during efficient, pop out visual search through suppression of the representation of the nontarget distractors. For a fraction of these neurons, the magnitude of this distractor suppression varied with the proximity of the target to the receptive field, exhibiting more suppression of the distractor representation when the target was nearby than when the target was distant. The purpose of this study was to determine whether the variation of distractor suppression related to target proximity varied with target-distractor feature similarity. The effect of target proximity on distractor suppression did not vary with target-distractor similarity and therefore may be an endogenous property of the selection process. PMID- 14749316 TI - Independent components of color natural scenes resemble V1 neurons in their spatial and color tuning. AB - It has been hypothesized that mammalian sensory systems are efficient because they reduce the redundancy of natural sensory input. If correct, this theory could unify our understanding of sensory coding; here, we test its predictions for color coding in the primate primary visual cortex (V1). We apply independent component analysis (ICA) to simulated cone responses to natural scenes, obtaining a set of colored independent component (IC) filters that form a redundancy reducing visual code. We compare IC filters with physiologically measured V1 neurons, and find great spatial similarity between IC filters and V1 simple cells. On cursory inspection, there is little chromatic similarity; however, we find that many apparent differences result from biases in the physiological measurements and ICA analysis. After correcting these biases, we find that the chromatic tuning of IC filters does indeed resemble the population of V1 neurons, supporting the redundancy-reduction hypothesis. PMID- 14749318 TI - Analysis of the optimal channel density of the squid giant axon using a reparameterized Hodgkin-Huxley model. AB - A reparameterized Hodgkin-Huxley-type model is developed that improves the 1952 model's fit to the biological action potential. In addition to altering Na(+) inactivation and K(+) activation kinetics, a voltage-dependent gating-current mechanism has been added to the model. The resulting improved model fits the experimental trace nearly exactly over the rising phase, and it has a propagation velocity that is within 3% of the experimentally measured value of 21.2 m/s (at 18.5 degrees C). Having eliminated most inaccuracies associated with the velocity dependent rising phase of the action potential, the model is used to test Hodgkin's maximum velocity hypothesis, which asserts that channel density has evolved to maximize conduction velocity. In fact the predicted optimal channel density is more than twice as high as the actual squid channel density. When the available capacitance is reduced to approximate more modern serial Na(+)-channel models, the optimal channel density is 4 times the actual value. We suggest that, although Hodgkin's maximum velocity hypothesis is acceptable as a first approximation, the microscopic optimization perspective of natural selection will not explain the channel density of the squid unless other constraints are taken into account, for example, the metabolic costs of velocity. PMID- 14749317 TI - Sodium along with low-threshold potassium currents enhance coincidence detection of subthreshold noisy signals in MSO neurons. AB - Voltage-dependent membrane conductances support specific neurophysiological properties. To investigate the mechanisms of coincidence detection, we activated gerbil medial superior olivary (MSO) neurons with dynamic current-clamp stimuli in vitro. Spike-triggered reverse-correlation analysis for injected current was used to evaluate the integration of subthreshold noisy signals. Consistent with previous reports, the partial blockade of low-threshold potassium channels (I(KLT)) reduced coincidence detection by slowing the rise of current needed on average to evoke a spike. However, two factors point toward the involvement of a second mechanism. First, the reverse correlation currents revealed that spike generation was associated with a preceding hyperpolarization. Second, rebound action potentials are 45% larger compared to depolarization-evoked spikes in the presence of an I(KLT) antagonist. These observations suggest that the sodium current (I(Na)) was substantially inactivated at rest. To test this idea, I(Na) was enhanced by increasing extracellular sodium concentration. This manipulation reduced coincidence detection, as reflected by slower spike-triggering current, and diminished the hyperpolarization phase in the reverse-correlation currents. As expected, a small outward bias current decreased the pre-spike hyperpolarization phase, and TTX blockade of I(Na) nearly eliminated the hyperpolarization phase in the reverse correlation current. A computer model including Hodgkin-Huxley type conductances for spike generation and for I(KLT) showed reduction in coincidence detection when I(KLT) was reduced or when I(Na) was increased. We hypothesize that desirable synaptic signals first remove some inactivation of I(Na) and reduce activation of I(KLT) to create a brief temporal window for coincidence detection of subthreshold noisy signals. PMID- 14749319 TI - On the relation between object shape and grasping kinematics. AB - Despite the many studies on the visual control of grasping, little is known about how and when small variations in shape affect grasping kinematics. In the present study we asked subjects to grasp elliptical cylinders that were placed 30 and 60 cm in front of them. The cylinders' aspect ratio was varied systematically between 0.4 and 1.6, and their orientation was varied in steps of 30 degrees. Subjects picked up all noncircular cylinders with a hand orientation that approximately coincided with one of the principal axes. The probability of selecting a given principal axis was the highest when its orientation was equal to the preferred orientation for picking up a circular cylinder at the same location. The maximum grip aperture was scaled to the length of the selected principal axis, but the maximum grip aperture was also larger when the length of the axis orthogonal to the grip axis was longer than that of the grip axis. The correlation between the grip aperture--or the hand orientation--at a given instant, and its final value, increased monotonically with the traversed distance. The final hand orientation could already be inferred from its value after 30% of the movement distance with a reliability that explains 50% of the variance. For the final grip aperture, this was only so after 80% of the movement distance. The results indicate that the perceived shape of the cylinder is used for selecting appropriate grasping locations before or early in the movement and that the grip aperture and orientation are gradually attuned to these locations during the movement. PMID- 14749320 TI - Contribution of intrinsic neuronal factors in the generation of cortically driven electrographic seizures. AB - Some electrographic seizures are generated intracortically. The cellular and ionic bases of cortically generated spontaneous seizures are not fully understood. Here we investigated spontaneously occurring seizures consisting of spike-wave complexes intermingled with fast runs in ketamine-xylazine anesthetized cats, using dual intracellular recordings in which one pipette contained a control solution and another pipette contained blockers of K(+), Na(+), or Ca(2+) currents. We show that closely located neocortical neurons display virtually identical fluctuations of the membrane potential during electrographic seizures, thus directly demonstrating a high degree of focal synchrony during paroxysmal activity. In addition to synaptic drives, the persistent Na(+) current [I(Na(p))] and probably the high-threshold Ca(2+) current contributed to the generation of paroxysmal depolarizing shifts (PDSs) during cortically driven seizures. Ca(2+)-activated K(+) current [I(K(Ca))] took also part in the control of the amplitude and duration of PDSs. The hyperpolarizing components of seizures largely depended on Cs(+)-sensitive K(+) currents. I(K(Ca)) played a significant, while not exclusive, role in the mediation of hyperpolarizing potentials related to EEG "waves" during spike-wave seizures. We conclude that intrinsic cellular factors have significant role in the generation of depolarizing and hyperpolarizing components of seizures. PMID- 14749321 TI - Improved spike-sorting by modeling firing statistics and burst-dependent spike amplitude attenuation: a Markov chain Monte Carlo approach. AB - Spike-sorting techniques attempt to classify a series of noisy electrical waveforms according to the identity of the neurons that generated them. Existing techniques perform this classification ignoring several properties of actual neurons that can ultimately improve classification performance. In this study, we propose a more realistic spike train generation model. It incorporates both a description of "nontrivial" (i.e., non-Poisson) neuronal discharge statistics and a description of spike waveform dynamics (e.g., the events amplitude decays for short interspike intervals). We show that this spike train generation model is analogous to a one-dimensional Potts spin-glass model. We can therefore tailor to our particular case the computational methods that have been developed in fields where Potts models are extensively used, including statistical physics and image restoration. These methods are based on the construction of a Markov chain in the space of model parameters and spike train configurations, where a configuration is defined by specifying a neuron of origin for each spike. This Markov chain is built such that its unique stationary density is the posterior density of model parameters and configurations given the observed data. A Monte Carlo simulation of the Markov chain is then used to estimate the posterior density. We illustrate the way to build the transition matrix of the Markov chain with a simple, but realistic, model for data generation. We use simulated data to illustrate the performance of the method and to show that this approach can easily cope with neurons firing doublets of spikes and/or generating spikes with highly dynamic waveforms. The method cannot automatically find the "correct" number of neurons in the data. User input is required for this important problem and we illustrate how this can be done. We finally discuss further developments of the method. PMID- 14749322 TI - Val133 and Cys137 in transmembrane segment 2 are close to Arg935 and Gly939 in transmembrane segment 11 of human P-glycoprotein. AB - P-glycoprotein (P-gp; ABCB1) transports a wide variety of structurally diverse compounds out of the cell. The protein has two homologous halves joined by a linker region. Each half consists of a transmembrane (TM) domain with six TM segments and a nucleotide-binding domain. The drug substrate-binding pocket is at the interface between the TM segments in each half of the protein. Preliminary studies suggested that the arrangement of the two halves of P-gp shows rotational symmetry (i.e. "head-to-tail" arrangement). Here, we tested this model by determining whether the cytoplasmic ends of TM2 and TM3 in the N-terminal half are in close contact with TM11 in the C-terminal half. Mutants containing a pair of cysteines in TM2/TM11 or TM3/TM11 were subjected to oxidative cross-linking with copper phenanthroline. Two of the 110 TM2/TM11 mutants, V133C(TM2)/G939C(TM11) and C137C(TM2)/A935C (TM11), were cross-linked at 4 degrees C, when thermal motion is reduced. Cross-linking was specific since no cross-linked product was detected in the 100 double Cys TM3/TM11 mutants. Vanadate trapping of nucleotide or the presence of some drug substrates inhibited cross-linking of mutants V133C(TM2)/G939C(TM11) and C137C(TM2)/A935C(TM11). Cross linking of TM2 and TM11 also blocked drug-stimulated ATPase activity. The close proximity of TM2/TM11 and TM5/TM8 (Loo, T. W., Bartlett, M. C., and Clarke, D. M. (2004) J. Biol. Chem. 279, 7692-7697) indicates that these regions between the two halves must enclose the drug-binding pocket at the cytoplasmic side of P-gp. They may form the "hinges" required for conformational changes during the transport cycle. PMID- 14749323 TI - Role of glycosylation in the organic anion transporter OAT1. AB - Organic anion transporters (OAT) play essential roles in the body disposition of clinically important anionic drugs, including antiviral drugs, antitumor drugs, antibiotics, antihypertensives, and anti-inflammatories. We reported previously (Kuze, K., Graves, P., Leahy, A., Wilson, P., Stuhlmann, H., and You, G. (1999) J. Biol. Chem. 274, 1519-1524) that tunicamycin, an inhibitor of asparagine linked glycosylation, significantly inhibited organic anion transport in COS-7 cells expressing a mouse organic anion transporter (mOAT1), suggesting an important role of glycosylation in mOAT1 function. In the present study, we investigated the effect of disrupting putative glycosylation sites in mOAT1 as well as its human counterpart, hOAT1, by mutating asparagine to glutamine and assessing mutant transporters in HeLa cells. We showed that the putative glycosylation site Asp-39 in mOAT1 was not glycosylated but the corresponding site (Asp-39) in hOAT1 was glycosylated. Disrupting Asp-39 resulted in a complete loss of transport activity in both mOAT1 and hOAT1 without affecting their cell surface expression, suggesting that the loss of function is not because of deglycosylation of Asp-39 per se but rather is likely because of the change of this important amino acid critically involved in the substrate binding. Single replacement of asparagines at other sites had no effect on transport activity indicating that glycosylation at individual sites is not essential for OAT function. In contrast, a simultaneous replacement of all asparagines in both mOAT1 and hOAT1 impaired the trafficking of the transporters to the plasma membrane. In summary, we provided the evidence that 1) Asp-39 is crucially involved in substrate recognition of OAT1, 2) glycosylation at individual sites is not required for OAT1 function, and 3) glycosylation plays an important role in the targeting of OAT1 onto the plasma membrane. This study is the first molecular identification and characterization of glycosylation of OAT1 and may provide important insights into the structure-function relationships of the organic anion transporter family. PMID- 14749324 TI - The role of a conserved acidic residue in calcium-dependent protein folding for a low density lipoprotein (LDL)-A module: implications in structure and function for the LDL receptor superfamily. AB - One common feature of the more than 1,000 complement-type repeats (or low density lipoprotein (LDL)-A modules) found in LDL receptor and the other members of the LDL receptor superfamily is a cluster of five highly conserved acidic residues in the C-terminal region, DXXXDXXDXXDE. However, the role of the third conserved aspartate of these LDL-A modules in protein folding and ligand recognition has not been elucidated. In this report, using a model LDL-A module and several experimental approaches, we demonstrate that this acidic residue, like the other four conserved acidic residues, is involved in calcium-dependent protein folding. These results suggest an alternative calcium coordination conformation for the LDL-A modules. The proposed model provides a plausible explanation for the conservation of this acidic residue among the LDL-A modules. Furthermore, the model can explain why mutations of this residue in human LDL receptor cause familial hypercholesterolemia. PMID- 14749325 TI - The surface coat of the mammal-dwelling infective trypomastigote stage of Trypanosoma cruzi is formed by highly diverse immunogenic mucins. AB - A thick coat of mucin-like glycoproteins covers the surface of Trypanosoma cruzi and plays a crucial role in parasite protection and infectivity and host immunomodulation. The appealing candidate genes coding for the mucins of the mammal-dwelling stages define a heterogeneous family termed TcMUC, which comprises up to 700 members, thus precluding a genetic approach to address the protein core identity. Here, we demonstrate by multiple approaches that the TcMUC II genes code for the majority of trypomastigote mucins. These molecules display a variable, non-repetitive, highly O-glycosylated central domain, followed by a short conserved C terminus and a glycosylphosphatidylinositol anchor. A simultaneous expression of multiple TcMUC II gene products was observed. Moreover, the C terminus of TcMUC II mucins, but not their central domain, elicited strong antibody responses in patients with Chagas' disease and T. crusi infected animals. This highly diverse coat of mucins may represent a refined parasite strategy to elude the mammalian host immune system. PMID- 14749326 TI - A novel endonucleolytic mechanism to generate the CCA 3' termini of tRNA molecules in Thermotoga maritima. AB - The tRNA 3'-terminal CCA sequence is essential for aminoacylation of the tRNAs and for translation on the ribosome. The tRNAs are transcribed as larger precursor molecules containing 5' and 3' extra sequences. In the tRNAs that do not have the encoded CCA, the 3' extra sequence after the discriminator nucleotide is usually cleaved off by the tRNA 3' processing endoribonuclease (3' tRNase, or RNase Z), and the 3'-terminal CCA residues are added thereto. Here we analyzed Thermotoga maritima 3' tRNase for enzymatic properties using various pre tRNAs from T. maritima, in which all 46 tRNA genes encode CCA with only one exception. We found that the enzyme has the unprecedented activity that cleaves CCA-containing pre-tRNAs precisely after the CCA sequence, not after the discriminator. The assays for pre-tRNA variants suggest that the CA residues at nucleotides 75 and 76 are required for the enzyme to cleave pre-tRNAs after A at nucleotide 76 and that the cleavage occurs after nucleotide 75 if the sequence is not CA. Intriguingly, the pre-tRNA(Met) that is the only T. maritima pre-tRNA without the encoded CCA was cleaved after the discriminator. The kinetics data imply the existence of a CCA binding domain in T. maritima 3' tRNase. We also identified two amino acid residues critical for the cleavage site selection and several residues essential for the catalysis. Analysis of cleavage sites by 3' tRNases from another eubacteria Escherichia coli and two archaea Thermoplasma acidophilum and Pyrobaculum aerophilum corroborates the importance of the two amino acid residues for the cleavage site selection. PMID- 14749327 TI - Post-translational formylglycine modification of bacterial sulfatases by the radical S-adenosylmethionine protein AtsB. AB - C(alpha)-Formylglycine (FGly) is the catalytic residue of sulfatases. FGly is generated by post-translational modification of a cysteine (prokaryotes and eukaryotes) or serine (prokaryotes) located in a conserved (C/S)XPXR motif. AtsB of Klebsiella pneumoniae is directly involved in FGly generation from serine. AtsB is predicted to belong to the newly discovered radical S-adenosylmethionine (SAM) superfamily. By in vivo and in vitro studies we show that SAM is the critical co-factor for formation of a functional AtsB.SAM.sulfatase complex and for FGly formation by AtsB. The SAM-binding site of AtsB involves (83)GGE(85) and possibly also a juxtaposed FeS center coordinated by Cys(39) and Cys(42), as indicated by alanine scanning mutagenesis. Mutation of these and other conserved cysteines as well as treatment with metal chelators fully impaired FGly formation, indicating that all three predicted FeS centers are crucial for AtsB function. It is concluded that AtsB oxidizes serine to FGly by a radical mechanism that is initiated through reductive cleavage of SAM, thereby generating the highly oxidizing deoxyadenosyl radical, which abstracts a hydrogen from the serine-C(beta)H(2)-OH side chain. PMID- 14749328 TI - Targeted inhibition of p38 mitogen-activated protein kinase antagonizes cardiac injury and cell death following ischemia-reperfusion in vivo. AB - The p38 branch of the mitogen-activated protein kinase (MAPK) signaling cascade has been implicated as a regulator of cardiomyocyte apoptosis in culture as well as in the adult heart. However, considerable disagreement persists as to the functional effects attributed to p38 signaling, given that both pro- and anti apoptotic regulatory roles have been reported. To address this area of uncertainty in the literature, we investigated the cell death effects associated with p38 inactivation in both cultured neonatal cardiomyocytes and the adult heart. In vitro, adenoviral-mediated gene transfer of two different dominant negative-encoding p38 vectors reduced apoptosis induced by 2-deoxyglucose treatment, whereas overexpression of wild-type p38alpha or an activated mitogen activated protein kinase kinase (MKK)6 mutant each enhanced cell death. In vivo, transgenic mice expressing a dominant-negative MKK6 mutant or a dominant-negative p38alpha mutant were each significantly protected from ischemia-reperfusion injury, as assessed by infarct area measurements, DNA laddering, terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling, and functional assessment of ventricular performance. Similarly, transgenic mice overexpressing the p38-inactivating dual specificity phosphatase MAPK phosphatase-1 (MKP-1) were also partially protected, whereas MKP-1 gene-targeted mice showed greater injury after ischemia-reperfusion injury. Mechanistically, inhibition of p38 signaling promoted a dramatic up-regulation of Bcl-2 in the hearts of transgenic mice. In primary neonatal cardiomyocyte cultures, adenoviral-mediated gene transfer of a p38 inhibitory mutant up-regulated Bcl-2, whereas expression of an activated p38 mutant down-regulated Bcl-2 protein levels. Collectively, these results indicate that p38 functions as a pro-death signaling effector in both cultured myocytes as well as in the intact heart. PMID- 14749329 TI - Streptococcus pyogenes fibronectin-binding protein F2: expression profile, binding characteristics, and impact on eukaryotic cell interactions. AB - Some Streptococcus pyogenes (group A streptococci, GAS) strains have previously been shown to express the fibronectin-binding protein F2 instead of the functionally related but structurally dissimilar protein F1/SfbI. In this study, recombinant N-terminal and C-terminal portions and the two fibronectin-binding domains of protein F2 were used to assess affinity parameters of the interaction with fibronectin and its N-terminal 70-, 30-, and 45-kDa fragments. The association and dissociation equilibrium constants for both binding domains were in the nanomolar range, although the repeat domain of protein F2 exceeded the affinity of the unique domain by up to one order magnitude. Both domains primarily interacted with the 30-kDa fibronectin fragment. Using a prtF2 gene isogenic mutant of a serotype M49 GAS strain that does not harbor the protein F1/SfbI gene, the attachment values of whole bacteria to immobilized fibronectin and to HEp-2 epithelial cells were found to be 6- and 2-fold decreased, respectively. Reduction of prtF2 mutant internalization rates for eukaryotic cells exceeded the reduction of attachment rates, indicating an independent contribution of protein F2 to both processes. The prtF2 transcription and protein F2 expression profiles documented maximum expression at the transition to the stationary phase especially under aerobic growth condition. The protein F2 function as the major fibronectin-binding adhesin in a subset of GAS strains, its expression pattern, and highly specific interaction with fibronectin would be consistent with a status as an indispensable virulence factor for both earlier and later pathogenetic stages of GAS superficial infections. PMID- 14749330 TI - Identification of molecular intermediates in the assembly pathway of the MUC5AC mucin. AB - MUC5AC mucins secreted by HT-29 cells in culture are oligomeric glycoproteins with characteristics similar to the MUC5AC mucins isolated from human airway sputum (Sheehan, J. K., Brazeau, C., Kutay, S., Pigeon, H., Kirkham, S., Howard, M., and Thornton, D. J. (2000) Biochem. J. 347, 37-44). Therefore we have used this cell line as a model system to investigate the biosynthesis of this major airway mucin. Initial experiments showed that the MUC5AC mucins isolated from the cells were liable to depolymerization depending on the conditions used for their solubilization. Prevention against reduction resulted in large oligomers associated with the cells, similar to those secreted into the medium. Using a combination of density gradient centrifugation and agarose gel electrophoresis coupled with probes specific for different forms of the mucin we identified five major intracellular populations of the MUC5AC polypeptide (unglycosylated monomer and dimer, GalNAc-substituted dimer, fully glycosylated dimer, and higher order oligomers). Pulse-chase studies were performed to follow the flow of radioactivity through these various intracellular forms into the mature oligomeric mucin secreted into the medium (a process taking approximately 2-4 h). The results show that the mucin polypeptide undergoes dimerization and then becomes substituted with GalNAc residues prior to glycan elaboration to produce a mature mucin dimer, which then undergoes multimerization. These data indicate that this oligomeric mucin follows a similar assembly to the von Willebrand factor glycoprotein to yield long linear disulfide-linked chains. PMID- 14749331 TI - Chemical cross-linking and mass spectrometric identification of sites of interaction for UreD, UreF, and urease. AB - Synthesis of active Klebsiella aerogenes urease requires four accessory proteins to generate, in a GTP-dependent process, a dinuclear nickel active site with the metal ions bridged by a carbamylated lysine residue. The UreD and UreF accessory proteins form stable complexes with urease apoprotein, comprised of UreA, UreB, and UreC. The sites of protein-protein interactions were explored by using homobifunctional amino group-specific chemical cross-linkers with reactive residues being identified by matrix-assisted laser desorption-ionization time-of flight mass spectrometry (MALDI-TOF MS) of tryptic peptides. On the basis of studies of the UreABCD complex, UreD is capable of cross-linking with UreB Lys(9), UreB Lys(76), and UreC Lys(401). Furthermore UreD appears to be positioned over UreC Lys(515) according to decreased reactivity of this residue compared with its reactivity in UreD-free apoprotein. Several UreB-UreC and UreC UreC cross-links also were observed within this complex; e.g. UreB Lys(76) with the UreC amino terminus, UreB Lys(9) with UreC Lys(20), and UreC Lys(515) with UreC Lys(89). These interactions are consistent with the proximate surface locations of these residues observed in the UreABC crystal structure. MALDI-TOF MS analyses of UreABCDF are consistent with a cross-link between the UreF amino terminus and UreB Lys(76). On the basis of an unexpected cross-link between UreB Lys(76) and UreC Lys(382) (distant from each other in the UreABC structure) along with increased side chain reactivities for UreC Lys(515) and Lys(522), UreF is proposed to induce a conformational change within urease that repositions UreB and potentially could increase the accessibility of nickel ions and CO(2) to residues that form the active site. PMID- 14749332 TI - Alternative polyadenylation of adeno-associated virus type 5 RNA within an internal intron is governed by the distance between the promoter and the intron and is inhibited by U1 small nuclear RNP binding to the intervening donor. AB - Adeno-associated virus type 5 is unique among adeno-associated virus serotypes in that it uses a polyadenylation site in the center of the genome. The great majority of transcripts generated from the upstream P7 and P19 promoters are polyadenylated at a site in the central intron ((pA)p); however, most of the viral transcripts generated by the proximal P41 promoter are polyadenylated at the distal polyadenylation site at the 3' end of the genome (pA)d and subsequently spliced. Polyadenylation at (pA)p increases as the distance between the RNA initiation site and the intron and (pA)p site is increased. The steady state level of RNAs polyadenylated at (pA)p is independent of the promoter used or of the intervening sequence but is dependent upon competition with splicing, inhibition by U1 snRNP binding to the intron donor, and the intrinsic efficiency of the cleavage/polyadenylation reaction. Each of these determinants shows a marked dependence on the distance between the RNA initiation site and the intron and (pA)p. Finally, unlike other reported systems, inhibition of (pA)p by U1 snRNP binding to the intron donor is decreased as the distance between the donor and (pA)p is increased. PMID- 14749333 TI - Oxidative folding of Amaranthus alpha-amylase inhibitor: disulfide bond formation and conformational folding. AB - Oxidative folding is the fusion of native disulfide bond formation with conformational folding. This complex process is guided by two types of interactions: first, covalent interactions between cysteine residues, which transform into native disulfide bridges, and second, non-covalent interactions giving rise to secondary and tertiary protein structure. The aim of this work is to understand both types of interactions in the oxidative folding of Amaranthus alpha-amylase inhibitor (AAI) by providing information both at the level of individual disulfide species and at the level of amino acid residue conformation. The cystine-knot disulfides of AAI protein are stabilized in an interdependent manner, and the oxidative folding is characterized by a high heterogeneity of one , two-, and three-disulfide intermediates. The formation of the most abundant species, the main folding intermediate, is favored over other species even in the absence of non-covalent sequential preferences. Time-resolved NMR and photochemically induced dynamic nuclear polarization spectroscopies were used to follow the oxidative folding at the level of amino acid residue conformation. Because this is the first time that a complete oxidative folding process has been monitored with these two techniques, their results were compared with those obtained at the level of an individual disulfide species. The techniques proved to be valuable for the study of conformational developments and aromatic accessibility changes along oxidative folding pathways. A detailed picture of the oxidative folding of AAI provides a model study that combines different biochemical and biophysical techniques for a fuller understanding of a complex process. PMID- 14749334 TI - Bacillus subtilis CheC and FliY are members of a novel class of CheY-P hydrolyzing proteins in the chemotactic signal transduction cascade. AB - Rapid restoration of prestimulus levels of the chemotactic response regulator, CheY-P, is important for preparing bacteria and archaea to respond sensitively to new stimuli. In an extension of previous work (Szurmant, H., Bunn, M. W., Cannistraro, V. J., and Ordal, G. W. (2003) J. Biol. Chem. 278, 48611-48616), we describe a new family of CheY-P phosphatases, the CYX family, that is widespread among the bacteria and archaea. These proteins provide another pathway, in addition to the ones involving CheZ of the gamma- and beta-proteobacteria (e.g. Escherichia coli) or the alternative CheY that serves as a "phosphate sink" among the alpha-proteobacteria (e.g. Sinorhizobium meliloti), for dephosphorylating CheY-P. In particular, we identify CheC, known previously to be involved in adaptation to stimuli in Bacillus subtilis, as a CheY-P phosphatase. Using an in vitro assay used previously to demonstrate that the switch protein FliY is a CheY P phosphatase, we have shown that increasing amounts of CheC accelerate the hydrolysis of CheY-P. In vivo, a double mutant lacking cheC and the region of fliY that encodes the CheY-P binding domain is almost completely smooth swimming, implying that these cells contain very high levels of CheY-P. CheC appears to be primarily involved in restoring normal CheY-P levels following the addition of attractant, whereas FliY seems to act on CheY-P constitutively. The activity of CheC is relatively low compared to that of FliY, but we have shown that the chemotaxis protein CheD enhances the activity of CheC 5-fold. We suggest a model for how FliY, CheC, and CheD work together to regulate CheY-P levels in the bacterium. PMID- 14749335 TI - Role of vav1- and src-related tyrosine kinases in macrophage activation by CpG DNA. AB - Macrophage activation by CpG DNA requires toll-like receptor 9 and the adaptor protein MyD88. Gram-negative bacterial lipopolysaccharide also activates macrophages via a toll-like receptor pathway (TLR-4), but we and others have reported that lipopolysaccharide also stimulates tyrosine phosphorylation in macrophages. Herein we report that exposure of RAW 264.7 murine macrophages to CpG DNA (but not non-CpG DNA) provoked the rapid tyrosine phosphorylation of vav1. PP1, a selective inhibitor of src-related tyrosine kinases, blocked both the CpG DNA-mediated tyrosine phosphorylation of vav1 and the CpG DNA-mediated up regulation of macrophage tumor necrosis factor secretion and inducible nitric oxide synthase protein accumulation. Furthermore, we found that the inducible expression of any of three dominant interfering mutants of vav1 (a truncated protein, vavC; a form containing a point mutation in the regulatory tyrosine residue, vavYF174; and a form with an in-frame deletion of six amino acids required for the guanidine nucleotide exchange factor (GEF) activity of vav1 for rac family GTPases, vavGEFmt) consistently inhibited CpG DNA-mediated up regulation of tumor necrosis factor secretion and inducible nitric-oxide synthase protein accumulation in RAW-TT10 macrophages. Finally, we determined that CpG DNA mediated up-regulation of NF-kappaB activity (but not mitogen-activated protein kinase activation) was inhibited by preincubation with PP1 or by expression of the truncated vavC mutant. Taken together, our results indicate that the tyrosine phosphorylation of vav1 by a src-related tyrosine kinase or kinases plays an important role in the macrophage response to CpG DNA. PMID- 14749336 TI - Isolation and characterization of a novel trans-factor for luteinizing hormone receptor mRNA from ovary. AB - Post-transcriptional mechanisms play a major role in regulating luteinizing hormone (LH) receptor mRNA expression in the ovary. An ovarian cytosolic protein that we have identified in rats and humans, which binds to a polypyrimidine-rich bipartitate sequence in the coding region of LHR mRNA, acts as a trans-acting factor in this process. In the present study, we isolated and characterized this LH receptor mRNA-binding protein (LRBP) from rat ovary. LRBP was purified to homogeneity by cation exchange chromatography followed by Northwestern analysis and subsequent elution of the single protein band from SDS-polyacrylamide gel. Purified LRBP was subjected to N-terminal microsequencing followed by homology search, which revealed its identity as mevalonate kinase. Purified rat mevalonate kinase antibody recognized the gel-purified LRBP on Western blots performed with one- and two-dimensional SDS-polyacrylamide gels. When recombinant mevalonate kinase produced in human embryonic kidney cells (293 cells) was tested, it showed all of the characteristics of LRBP with respect to specificity of LHR mRNA binding sequence, as examined by gel mobility shift analysis. Inhibition of LHR mRNA binding activity of mevalonate kinase in the presence of ATP and mevalonate indicates that the RNA recognition site of mevalonate kinase might involve the ATP/mevalonate binding region of the protein. Treatment of 293 cells with mevastatin to deplete cellular mevalonate resulted in an increase in LHR mRNA binding activity of mevalonate kinase. Collectively, the data support the novel function of rat mevalonate kinase as a LHR mRNA-binding protein in the post transcriptional regulation of LH receptor expression in the ovary. PMID- 14749337 TI - Involvement of the junctional adhesion molecule-1 (JAM1) homodimer interface in regulation of epithelial barrier function. AB - Junctional adhesion molecule-1 (JAM1) is a tight junction-associated immunoglobulin superfamily protein implicated in the regulation of tight junctions and leukocyte transmigration. The structural basis for the function of JAM1 has yet to be determined. Here we provide evidence that JAM1 homodimer formation is important for its function in epithelial cells. Experiments were conducted to determine the effects of a panel of JAM1 monoclonal antibodies on epithelial barrier recovery after transient disruption by calcium switch. Two monoclonal antibodies were observed to inhibit barrier recovery in contrast to another monoclonal antibody that had no effect. Epitope mapping by phage display revealed that both inhibitory antibodies bind to a region of JAM1 located within the N-terminal Ig-like loop (residues 111-123). Competition experiments with synthetic peptides and site-directed mutagenesis confirmed the location of this epitope. Analysis of the crystal structure of JAM1 revealed that this epitope includes residues within the putative homodimer interface, and one of the two inhibitory antibodies was then shown to block JAM1 homodimer formation in vitro. Finally, mutations within the homodimer interface were shown to prevent enrichment of JAM1 at points of cell contact, presumably by interference with homophilic interactions. These findings suggest that homodimer formation may be important for localization of JAM1 at tight junctions and for regulation of epithelial barrier function. PMID- 14749338 TI - Survival motor neuron (SMN) protein interacts with transcription corepressor mSin3A. AB - Spinal muscular atrophy (SMA) is the leading genetic cause of infant mortality. SMA results from loss of survival motor neuron (SMN) expression and subsequent death of motor neuron cells. To study SMN-associated proteins that may be involved in transcriptional regulation, we carried out immunoprecipitation experiments and found that the transcription corepressor mSin3A associates with SMN protein. Deletional analysis localized the mSin3A-interacting domain to the exon 6 region of SMN. When targeted to a promoter, wild-type SMN was able to repress transcription of a downstream luciferase reporter gene. This repression was relieved by treatment with the histone deacetylase inhibitor trichostatin A in a dose-dependent manner, and deletion of exon 6 abolished the ability of SMN to repress the reporter gene. Analysis of SMN missense mutations within the exon 6 region implicated the SMA-associated mutation Y272C with impairment of the mSin3A-interaction. Gel filtration experiments revealed that wild-type SMN, via the exon 6 region, forms protein supra-complexes exceeding 40,000 kDa in size, whereas the Y272C mutation may affect higher order protein assembly, as the mutant SMN was more abundant in smaller complexes. Together, these findings provide a potential mechanism by which lack of fully functional SMN protein is detrimental to motor neuron survival. PMID- 14749339 TI - An assessment of the risks associated with the use of antibiotic resistance genes in genetically modified plants: report of the Working Party of the British Society for Antimicrobial Chemotherapy. AB - Development of genetically modified (GM) plants is contentious, in part because bacterial antibiotic resistance (AR) genes are used in their construction and often become part of the plant genome. This arouses concern that cultivation of GM plants might provide a reservoir of AR genes that could power the evolution of new drug-resistant bacteria. We have considered bacterial DNA transfer systems (conjugation, transduction and transformation) and mechanisms of recombination (homologous recombination, transposition, site-specific recombination and DNA repair) that together might productively transfer AR genes from GM plants to bacterial cells, but are unable to identify a credible scenario whereby new drug resistant bacteria would be created. However, we cannot entirely rule out the possibility of rare transfer events that involve novel mechanisms. Hence, we also considered if occasional transfers of AR genes (bla(TEM), aph(3'), aadA) from GM plants into bacteria would pose a threat to public health. These AR genes are common in many bacteria and each is found on mobile genetic elements that have moved extensively between DNA molecules and bacterial cells. This gene mobility has already severely compromised clinical use of antibiotics to which resistance is conferred. Accordingly, the argument that occasional transfer of these particular resistance genes from GM plants to bacteria would pose an unacceptable risk to human or animal health has little substance. We conclude that the risk of transfer of AR genes from GM plants to bacteria is remote, and that the hazard arising from any such gene transfer is, at worst, slight. PMID- 14749340 TI - Contribution of integrons, and SmeABC and SmeDEF efflux pumps to multidrug resistance in clinical isolates of Stenotrophomonas maltophilia. AB - OBJECTIVES: The contribution of integrons and efflux pumps to multidrug resistance in Stenotrophomonas maltophilia was evaluated. MATERIALS AND METHODS: Ninety-three S. maltophilia clinical isolates were studied. PCR and direct sequencing were used to detect the presence of integrons. Real-time PCR was performed to assess and quantify the expression of the Sme efflux pumps of S. maltophilia. RESULTS: Class 1 integrons were detected in 22% of clinical isolates and carried cassettes conferring resistance mainly to aminoglycosides and trimethoprim. The small multidrug resistance gene, smr, was found on class 1 integrons in six isolates. Thirty-one percent of the isolates overexpressed the smeDEF gene, as compared with a control strain, and 59% overexpressed the smeABC gene. Extrusion of ciprofloxacin and meropenem was specific to the SmeABC and SmeDEF pumps, respectively. CONCLUSION: SmeABC and SmeDEF efflux pumps play important roles in resistance of S. maltophilia to ciprofloxacin and meropenem. PMID- 14749341 TI - Pseudomonas aeruginosa strains harbouring an unusual blaVIM-4 gene cassette isolated from hospitalized children in Poland (1998-2001). AB - OBJECTIVES: During 1997-2001, 151 isolates of imipenem-resistant Pseudomonas aeruginosa were obtained from clinical specimens taken from children hospitalized in Warsaw, Poland. These strains were investigated further to determine the mechanism of resistance. METHODS: The strains were analysed by a combination of genotyping and PCR-based strategies. RESULTS: Eleven of these strains were found to contain the metallo-beta-lactamase (M beta L) gene bla(VIM-4). The first strain appeared in 1998, and P. aeruginosa strains harbouring this M beta L have become endemic in this hospital since then. All P. aeruginosa strains belonged to serotype O:6, and PFGE analysis revealed four different patterns and three sub types. All 11 M beta L-producing strains contained an identical class 1 integron with the usual 5' and 3' conserved sequences. The integron included two resistance cassettes, aacA4 in the first position and the bla(VIM-4) cassette in the second position. The bla(VIM-4) gene included an unusual direct repeat of 169 bp of the 3' portion of the bla(VIM-4) gene. CONCLUSIONS: An unusual bla(VIM-4) M beta L has become endemic in P. aeruginosa isolates infecting Polish children hospitalized on surgical wards. The formation of this unusual bla(VIM-4) gene cassette could be explained by a mechanism involving deletion of a segment of an ancestral tandem repeat of bla(VIM-4) via slipped strand replication, mediated by a combination of polymerase and integrase. PMID- 14749342 TI - Duplicate publication: a cautionary tale. PMID- 14749343 TI - Caspofungin modulates in vitro adherence of Candida albicans to plastic coated with extracellular matrix proteins. AB - OBJECTIVES: Some manifestations of candidiasis are associated with the formation of biofilms on inert or biological surfaces and the intrinsic resistance of Candida albicans biofilms to the most commonly used antifungal agents has been demonstrated. In this study, we report on the influence of the growth of C. albicans in medium containing a sub-inhibitory concentration (MIC/2) of caspofungin, on subsequent fungal adherence to plastic coated with extracellular matrix (ECM) proteins. METHODS: Eleven strains of C. albicans were studied: six strains were susceptible to fluconazole in vitro and five strains were resistant to this antifungal agent. RESULTS: Caspofungin induced a decrease in the adherence of all the tested strains that were susceptible to fluconazole but induced a decrease in the adherence of only 60% of the fluconazole-resistant strains. CONCLUSIONS: This study demonstrated the anti-adherent activity of caspofungin but indicated a reduced effect in the case of in vitro fluconazole resistance. These results indicated a possible relationship between the efficiency of caspofungin to inhibit the first step of the development of C. albicans biofilm and the resistance of C. albicans to fluconazole in vitro. PMID- 14749344 TI - Use of administrative healthcare claims to examine the effectiveness of trimethoprim-sulfamethoxazole versus fluoroquinolones in the treatment of community-acquired acute pyelonephritis in women. AB - OBJECTIVE: To evaluate the effectiveness of trimethoprim-sulfamethoxazole and fluoroquinolones in the treatment of community-acquired acute pyelonephritis. PATIENTS AND METHODS: We identified a population-based cohort of non-pregnant women aged 18-65 years, initially treated with trimethoprim-sulfamethoxazole or a fluoroquinolone for community-acquired pyelonephritis in an ambulatory care setting. Subjects were identified from a healthcare claims database in Manitoba, Canada for the period 15 February 1996 to 31 March 1999. Subsequent treatment failure, as evidenced by the provision of additional treatment up to 42 days post diagnosis, was compared between the two treatments. RESULTS: A total of 1084 women met inclusion criteria: 653 (60.2%) treated with trimethoprim sulfamethoxazole and 431 (39.8%) treated with a fluoroquinolone. Treatment outcomes were affected by subject age. At age 20, treatment with a fluoroquinolone resulted in a reduced probability of treatment failure compared with trimethoprim-sulfamethoxazole (odds ratio, 0.56; 95% CI, 0.33-0.97). At age 60, there was no difference in the probability of treatment failure (odds ratio, 1.61; 95% CI, 0.82-3.16). No other subject characteristics impacted comparative effectiveness; however, several characteristics increased the odds of treatment failure irrespective of the initial antibiotic. These included: recent urinary tract infection (odds ratio, 2.07; 95% CI, 1.14-3.57), recent antibiotic use (odds ratio, 1.40; 95% CI, 1.00-1.96;), and a treatment duration of less than 10 days (odds ratio, 2.18; 95% CI, 1.59-2.99). CONCLUSION: Younger subjects ( approximately 20 years) treated with fluoroquinolones were less likely to experience treatment failure than those treated with trimethoprim sulfamethoxazole. Treatment durations of less than 10 days resulted in a higher probability of treatment failure regardless of the initial antibiotic. PMID- 14749345 TI - Surveillance of HIV antiretroviral drug resistance in treated individuals in England: 1998-2000. AB - OBJECTIVES: To establish a surveillance programme for HIV drug resistance within the UK covering the years from 1998 to 2000, following the introduction of triple combination antiretroviral therapy. METHODS: Sentinel sites included large, medium sized and small clinical centres. Data were analysed until December 2000. RESULTS: Of nearly 300 samples tested, results from 91, 92 and 92 patients, respectively in 1998, 1999 and 2000, who were receiving HIV therapy with a viral load >2000 copies/mL, the majority had viruses with some degree of drug resistance. Overall, the presence of any resistance increased between 1998 and 1999, and fell again in 2000 (69% versus 88% versus 55%). However, major differences were observed between drug classes, such that non-nucleoside analogue reverse transcriptase inhibitor (NNRTI) resistance rose dramatically over the period studied. We show that this correlated with increased NNRTI prescribing. Furthermore, an overall increase in prevalence of viruses with resistance to one or more drugs within all three available classes was observed. A higher prevalence of drug resistance was observed in patients from smaller clinical centres. CONCLUSIONS: This is the first such sentinel surveillance dataset from the UK, and is unique in correlating these data with national antiretroviral prescribing patterns. Our findings are relevant to the increased transmission of HIV drug resistance observed over this period. PMID- 14749346 TI - Effect of norepinephrine on cefpirome tissue concentrations in healthy subjects. AB - OBJECTIVES: To test whether norepinephrine (NOR) affects tissue microcirculation and impairs plasma-to-tissue equilibration of antimicrobial agents. MATERIALS AND METHODS: Eight healthy male volunteers were enrolled to an analyst-blinded, randomized, two-period two-sequence crossover study. A single intravenous dose of 2 g of cefpirome was administered simultaneously with starting a continuous infusion of NOR (0.16 microg/kg per min) or placebo (PL) over 180 min. The microdialysis technique was used for the assessment of unbound cefpirome concentrations in skeletal muscle tissue and subcutaneous adipose tissue. Free plasma concentrations were related to corresponding tissue concentrations. Haemodynamics were determined by the measurement of mean arterial blood pressure (MAP), heart rate and forearm blood flow (FBF). RESULTS: Area under the concentration-time-curve (AUC) values of cefpirome for interstitium and plasma were not significantly different between the PL and NOR groups (P > 0.47). Tissue penetration of cefpirome as described by the ratios of the AUCs from 0 to 180 min for tissue to the AUC values for plasma were 0.81 +/- 0.34 for the PL group and 0.80 +/- 0.26 for the NOR group (P > 0.05). Baseline values of MAP, heart rate and FBF were not significantly different between study days. MAP increased significantly following NOR administration from 73.3 +/- 3.5 mmHg at baseline to 94.0 +/- 5.2 mmHg during infusion (P = 0.017). NOR exerted no significant effects on FBF. CONCLUSIONS: We have shown that intravenous administration of NOR does not exert a significant effect on peripheral blood flow and tissue penetration of cefpirome in healthy men. This might be attributed to systemic regulatory mechanisms, which probably fully compensate for major changes in blood flow in peripheral tissues. PMID- 14749348 TI - Unique gene expression signatures of independently-derived human embryonic stem cell lines. AB - Human embryonic stem cells (hESCs) have the potential to differentiate to diverse cell types. This ability endows hESCs with promise for the development of novel therapeutics, as well as promise for the development of a rigorous genetic system to probe human gene function. However, in spite of the impending utility of hESCs for clinical and basic applications, little is known about their fundamental properties. Recent reports have documented transcriptional profiles of mouse embryonic stem cells (mESCs), adult stem cells and a single hESC line, H9. To date, however, the transcriptional profiles of independently-derived hESC lines have not been compared. In order to examine the similarities and differences in multiple hESC lines, we compared gene expression profiles of the HSF-1, HSF-6 and H9 lines. We found that the majority of genes examined were expressed in all three cell lines. However, we also observed that each line possessed a unique expression signature; the expression of many genes was limited to just one or two hESC lines. We suggest that the observed differences in gene expression between independently-derived hESC lines may reflect inherent differences in the initial culture of each line and/or the underlying genetics of the embryos from which the lines were derived. PMID- 14749347 TI - Molecular pathophysiology of mucolipidosis type IV: pH dysregulation of the mucolipin-1 cation channel. AB - Mucolipidosis type IV (MLIV) is an autosomal recessive neurogenetic disorder characterized by developmental abnormalities of the brain and impaired neurological, ophthalmologic and gastric function. Large vacuoles accumulate in various types of cells in MLIV patients. However, the pathophysiology of the disease at the cellular level is still unknown. MLIV is caused by mutations in a recently described gene, MCOLN1, encoding mucolipin-1 (ML1), a 65 kDa protein whose function is also unknown. ML1 shows sequence homology and topological similarities with polycystin-2 and other transient receptor potential (Trp) channels. In this study, we assessed both, whether ML1 has ion channel properties, and whether disease-causing mutations in MCOLN1 have functional differences with the wild-type (WT) protein. ML1 channel function was assessed from endosomal vesicles of null (MCOLN1(-/-)) and ML1 over-expressing cells, and liposomes containing the in vitro translated protein. Evidence from both preparations indicated that WT ML1 is a multiple subconductance non-selective cation channel whose function is inhibited by a reduction of pH. The V446L and DeltaF408 MLIV causing mutations retain channel function but not the sharp inhibition by lowering pH. Atomic force imaging of ML1 channels indicated that changes in pH modified the aggregation of unitary channels. Mutant-ML1 did not change in size on reduction of pH. The data indicate that ML1 channel activity is regulated by a pH-dependent mechanism that is deficient in some MLIV causing mutations of the gene. The evidence also supports a novel role for cation channels in the acidification and normal endosomal function. PMID- 14749349 TI - Haplotypic analyses of the IGF2-INS-TH gene cluster in relation to cardiovascular risk traits. AB - The IGF2-INS-TH genomic region has been implicated in various common disorders including the metabolic syndrome, type 2 diabetes and coronary heart disease (CHD). Here we present detailed haplotype analysis of 2743 males 51-62 years old in relation to body weight and composition, blood pressure (BP) and plasma triglycerides (TG). Use of the total data set was complicated by the number of loci typed, missing data, multi-allelic markers and continuous trait phenotypes. Different algorithms and subsets of the data were analysed using the programmes haplotype trend regression, haplo.score, evolutionary-based haplotype analysis package and Phase, in conjunction with SPSS. Ten haplotypes designated in frequency order *1(20.0%) to *10(3.4%) represented 89% of all haplotypes. Haplotype *5 protected against obesity. Haplotype *4 carriers exhibited elevated BP and fat mass, haplotype *6 was associated with raised plasma TG levels. Haplotype *8 also showed similar magnitude effects as *4. These cohort trait analyses and detailed haplotypic analyses enable integration with published case data. Haplotypes *4, *6 and *8 are the only INS VNTR class III-bearing haplotypes, although differing in flanking haplotype, whereas *5 displays unique features in all three genes (with significant commonality with type 1 diabetes predisposition haplotypes). We propose that long repeat insertion in the insulin gene promoter ('class III'), reported to result in low insulin production, predisposes to the metabolic syndrome features of elevated BP, fat mass or TG level, therefore appearing more frequently in type 2 diabetic, polycystic ovary syndrome and CHD cases. The functional element(s) of *5 for weight-lowering could reside in any of the three genes. PMID- 14749350 TI - Differences in assembly or stability of complex I and other mitochondrial OXPHOS complexes in inherited complex I deficiency. AB - NADH-ubiquinone oxidoreductase (complex I) deficiency is amongst the most encountered defects of the mitochondrial oxidative phosphorylation (OXPHOS) system and is associated with a wide variety of clinical signs and symptoms. Mutations in complex I nuclear structural genes are the most common cause of isolated complex I enzyme deficiencies. The cell biological consequences of such mutations are poorly understood. In this paper we have used blue native electrophoresis in order to study how different nuclear mutations affect the integrity of mitochondrial OXPHOS complexes in fibroblasts from 15 complex I deficient patients. Our results show an important decrease in the levels of intact complex I in patients harboring mutations in nuclear-encoded complex I subunits, indicating that complex I assembly and/or stability is compromised. Different patterns of low molecular weight subcomplexes are present in these patients, suggesting that the formation of the peripheral arm is affected at an early assembly stage. Mutations in complex I genes can also affect the stability of other mitochondrial complexes, with a specific decrease of fully-assembled complex III in patients with mutations in NDUFS2 and NDUFS4. We have extended this analysis to patients with an isolated complex I deficiency in which no mutations in structural subunits have been found. In this group, we can discriminate between complex I assembly and catalytic defects attending to the fact whether there is a correlation between assembly/activity levels or not. This will help us to point more selectively to candidate genes for pathogenic mutations that could lead to an isolated complex I defect. PMID- 14749351 TI - The complex genetic epidemiology of prostate cancer. AB - Prostate cancer is the most frequent cancer among men in most developed countries, yet little is known about its causes. Older age, African ancestry and a positive family history of prostate cancer have long been recognized as important risk factors. The evidence that genetics probably plays a critical role is based on a variety of study designs, including case-control, cohort, twin and family-based, all of which are reviewed in detail. The search for prostate cancer susceptibility genes by linkage studies offered early hope that finding genes would be as 'easy' as finding genes for breast cancer and colon cancer susceptibilities. However, this hope has been dampened by the difficulty of replicating promising regions of linkage. This review provides updates on recent developments, and a broad view of the disparate findings from different linkage studies. Early linkage results have provided targeted candidate regions for prostate cancer susceptibility loci, including HPC1 on chromosome 1q23-25, PCAP on chromosome 1q42-43, CAPB on chromosome 1p36, linkage to chromosome 8p22-23, HPC2 on chromosome 17p, HPC20 on chromosome 20q13, and HPCX on chromosome Xq27 28. These linkage findings lead to refined mapping and mutation screening of several strong candidate genes, including ELAC2, RNASEL and MSR1. Up to now, a total of 10 genome-wide linkage scans for prostate cancer susceptibility have been completed, and are reviewed. Furthermore, recent findings that Gleason's grade, a measure of aggressiveness of prostate cancer, is linked to several genomic regions are reviewed. Finally, the roles of environmental and dietary risk factors, and common genetic polymorphisms of genes likely to play a role in common forms of prostate cancer, are briefly discussed within in the context of searching for genes that influence prostate cancer risk. PMID- 14749352 TI - Modeled microgravity inhibits osteogenic differentiation of human mesenchymal stem cells and increases adipogenesis. AB - Space flight-induced bone loss has been attributed to a decrease in osteoblast function, without a significant change in bone resorption. To determine the effect of microgravity (MG) on bone, we used the Rotary Cell Culture System [developed by the National Aeronautics and Space Administration (NASA)] to model MG. Cultured mouse calvariae demonstrated a 3-fold decrease in alkaline phosphatase (ALP) activity and failed to mineralize after 7 d of MG. ALP and osteocalcin gene expression were also decreased. To determine the effects of MG on osteoblastogenesis, we cultured human mesenchymal stem cells (hMSC) on plastic microcarriers, and osteogenic differentiation was induced immediately before the initiation of modeled MG. A marked suppression of hMSC differentiation into osteoblasts was observed because the cells failed to express ALP, collagen 1, and osteonectin. The expression of runt-related transcription factor 2 was also inhibited. Interestingly, we found that peroxisome proliferator-activated receptor gamma (PPARgamma2), which is known to be important for adipocyte differentiation, adipsin, leptin, and glucose transporter-4 are highly expressed in response to MG. These changes were not corrected after 35 d of readaptation to normal gravity. In addition, MG decreased ERK- and increased p38-phosphorylation. These pathways are known to regulate the activity of runt-related transcription factor 2 and PPARgamma2, respectively. Taken together, our findings indicate that modeled MG inhibits the osteoblastic differentiation of hMSC and induces the development of an adipocytic lineage phenotype. This work will increase understanding and aid in the prevention of bone loss, not only in MG but also potentially in age-and disuse-related osteoporosis. PMID- 14749353 TI - Insulin-like growth factor binding protein-6 transgenic mice: postnatal growth, brain development, and reproduction abnormalities. AB - In biological fluids, IGFs bind to six distinct binding proteins (IGFBP-1 to -6). IGFBP-6 is of particular interest because it has been shown to inhibit proliferation in many cell types and to be synthesized in the central nervous system (CNS). It also has the strongest affinity for IGF-II among the IGFBPs. To study IGFBP-6 function in vivo, we established IGFBP-6 transgenic mice in which human IGFBP-6 (hIGFBP-6) cDNA is expressed under the control of the glial fibrillary acidic protein (GFAP) promoter. Northern and Western blot analysis revealed strong transgene expression in the CNS. With histological examination of the CNS, cerebellum size and weight proved to be reduced by about 25% and 35%, respectively, and there were smaller numbers of differentiated, GFAP-expressing astrocytes than in wild-type mice. Between birth and 1 month of age, transgenic mice had high levels of circulating hIGFBP-6 and reduced plasma IGF-I, and, as a result, body weight was significantly reduced. Reproductive physiology was also affected. Litter size was reduced by 27% when wild-type males were mated with 3 month-old transgenic females and by 66% when mated with 6-month-old transgenic females. Histological examination of ovaries of transgenic mice revealed a marked decrease in weight and in the number of corpora lutea, suggesting altered ovulation, and circulating LH levels were reduced by 50%. Our results indicate that this new model of transgenic mouse may prove to be a useful tool in elucidating the in vivo role of IGFBP-6 in the brain, especially in regard to hypothalamic control, and in reproductive physiology. PMID- 14749354 TI - Synergistic induction of the nicotinamide adenine dinucleotide-linked 15 hydroxyprostaglandin dehydrogenase by an androgen and interleukin-6 or forskolin in human prostate cancer cells. AB - The nicotinamide adenine dinucleotide-dependent 15-hydroxyprostaglandin dehydrogenase (15-PGDH) catalyzes the oxidation of 15 (S)-hydroxyl group of prostaglandins and lipoxins and participates along with cyclooxygenases and lipoxygenases in controlling the cellular levels of prostaglandins and lipoxins. 15-PGDH could be induced by IL-6 and forskolin in addition to androgens in a time and dose-dependent manner but not by other cytokines and growth factors in LNCaP cells. Concurrent addition of IL-6 and forskolin showed additive effect in the induction of 15-PGDH activity. However, combined addition of dihydrotestosterone (DHT) and IL-6 or DHT plus forskolin exhibited synergistic induction of 15-PGDH activity. The increase in enzyme activity was correlated with the expression of the enzyme protein as shown by Western blot analysis. The induction by DHT or IL 6 or forskolin or their combinations was inhibited by antiandrogen, casodex, in a dose-dependent manner, indicating that a functional androgen receptor was required for the action of any of these three agents. The induction by forskolin plus DHT or by either agent or by IL-6 alone was greatly inhibited by H-89, indicating the involvement of protein kinase A in the actions of forskolin, DHT, and IL-6. The induction of 15-PGDH by IL-6 was also blocked by some other protein kinase inhibitors, indicating the participation of MAPK, MAPK/ERK kinase, and STAT3 in the signaling pathway of IL-6. These results indicate that the induction of 15-PGDH by DHT, IL-6, and forskolin in LNCaP cells may involve a functional androgen receptor and phosphorylation-dependent multiple signaling pathways. PMID- 14749355 TI - Aberrant cholesterol transport and impaired steroidogenesis in Leydig cells lacking estrogen sulfotransferase. AB - Estrogen sulfotransferase (EST) is a cytosolic enzyme that catalyzes the sulfoconjugation and inactivation of estrogens. It is expressed abundantly in the mammalian testes in which it may modulate the activity of locally produced estrogen. We demonstrate here that testicular Leydig cells from mice rendered deficient in EST expression by targeted gene deletion acquire a phenotype of increased cholesterol ester accumulation and impaired steroidogenesis with natural aging or in response to estrogen challenge. Abnormal accumulation of cholesterol ester in the mutant Leydig cells correlated with induced expression of the scavenger receptor type B class I, and cultured EST-deficient but not wild type Leydig cells avidly uptook high-density lipoprotein cholesterol ester ex vivo. EST-deficient Leydig cells in culture produced 50-70% less testosterone than wild-type cells. This deficiency was reversed by androstenedione but not progesterone supplementation, indicating that reduced activities of 17-alpha hydroxylase-17, 20-lyase were responsible. This conclusion was corroborated by decreased expression levels of 17-alpha-hydroxylase-17, 20-lyase but not of other key steroidogenic enzymes in the mutant cells. These results suggest that EST plays a physiologic role in protecting Leydig cells from estrogen-induced biochemical lesions and provide an example of critical regulation of tissue estrogen sensitivity by a ligand-transformation enzyme rather than through estrogen receptors. PMID- 14749356 TI - Inhibitory effect of C-type natriuretic peptide (CNP) on cultured cardiac myocyte hypertrophy: interference between CNP and endothelin-1 signaling pathways. AB - C-type natriuretic peptide (CNP) is known to play a role in the local regulation of vascular tone. We recently found that CNP is also produced by cardiac ventricular cells. However, its local effect on myocyte hypertrophy remains to be elucidated. The present study investigated the effects of CNP on cultured cardiac myocyte hypertrophy and the interaction between CNP and endothelin-1 (ET-1) signaling pathways. CNP attenuated basal and ET-1-augumented protein synthesis, atrial natriuretic peptide secretion, hypertrophy-related gene expression, GATA-4 and MEF-2 DNA binding activities, Ca(2+)/calmodulin-dependent kinase II activity, and ERK phosphorylation. CNP also inhibited ET-1-induced increase in intracellular Ca(2+) concentration. These effects of CNP were mimicked by a cGMP analog, 8-bromo cGMP. However, the inhibitory effects of CNP on the hypertrophic response of myocytes were significantly diminished at high concentrations of ET 1. Although CNP increased intracellular cGMP levels in myocytes, ET-1 suppressed CNP-induced cellular cGMP accumulation. A protein kinase C activator and Ca(2+) ionophore mimicked this suppressive effect of ET-1. We further examined the effect of CNP on the paracrine action of ET-1 secreted from cardiac nonmyocytes. CNP and 8-bromo cGMP significantly inhibited ET-1 secretion from nonmyocytes. Although nonmyocyte-conditioned medium increased the protein synthesis in myocytes through endogenous ET-1 action, this increase was significantly attenuated by pretreatment of nonmyocytes with CNP and 8-bromo cGMP. These findings demonstrate that CNP inhibits ET-1-induced cardiac myocyte hypertrophy via a cGMP-dependent mechanism, and conversely, ET-1 inhibits CNP signaling by a protein kinase C- and Ca(2+)-dependent mechanism, suggesting mutual interference between CNP and ET-1 signaling pathways. PMID- 14749357 TI - Tumor necrosis factor-alpha induces serum amyloid A3 in mouse granulosa cells. AB - TNF-alpha has significant inhibitory effects on steroidogenesis and folliculogenesis and is associated with several inflammatory responses. Because ovulation is an inflammatory reaction, the effects of TNF on the family of acute phase proteins in granulosa cells were investigated. Granulosa cells from immature mice at 28 d of age were cultured in the presence of 10 ng TNF/ml for 24 h. Serum amyloid A3 (SAA3), a main acute-phase protein, was induced by TNF in granulosa cells. The other isoforms of serum amyloid proteins SAA1, SAA2, and SAA4 were neither expressed in granulosa cells nor induced by TNF. TNF did not induce SAA3 mRNA in granulosa cells from TNF receptor type 1 (TNFR1) knockout mice, although SAA3 mRNA was induced within 3 h after TNF treatment in wild-type cells. Two SAA3 promoters, -617/+73 and -198/+73, were responsive to TNF and to p65, a component of the TNF signaling molecule nuclear factor (NF)-kappaB. The 106/+73 promoter of SAA3 lacking a NF-kappaB-like site was not responsive to TNF or p65. In granulosa cells from TNFR1 knockout mice, the SAA3 promoter (-198/+73) was responsive to transfection with the p65 component of NF-kappaB, but neither TNF treatment nor overexpression of the p50 component of NF-kappaB increased promoter activity. Similar results were observed in the murine ovarian granulosa tumor cell line (OV3121-1). Overexpression of the inhibitor of NF-kappaB (called IkappaB) blocked SAA3 promoter activity induced by TNF and by p65 in OV3121-1 cells. Closer analysis of deletion mutants of the SAA3 promoter revealed the necessity of a NF-kappaB like site for responsiveness to TNF in the OV3121-1 cells. TNF rapidly increased p65 in OV3121-1 nuclei when compared with controls not treated with TNF. TNF also increased phospho-IkB and SAA3 in whole-cell homogenates as determined by Western blots. Thus, TNF likely increased SAA3 promoter activity and protein by activating NF-kappaB signaling via TNFR1 in mouse granulosa cells. SAA3 is a novel gene in granulosa cells with yet unknown functions in the ovary. PMID- 14749359 TI - Insulin-like growth factor-I augments chondrocyte hypertrophy and reverses glucocorticoid-mediated growth retardation in fetal mice metatarsal cultures. AB - The study aims were to improve our understanding of the mechanisms of glucocorticoid-induced growth retardation at the growth plate and determine whether IGF-I could ameliorate the effects. Fetal mouse metatarsals were cultured for up to 10 d with dexamethasone (Dex; 10(-6) m) and/or IGF-I and GH (both at 100 ng/ml). Both continuous and alternate-day Dex treatment inhibited bone growth to a similar degree, whereas IGF-I alone or together with Dex caused an increase in bone growth. GH had no effects. These observations may be explained at the cellular level; cell proliferation within the growing bone was decreased by Dex and increased by IGF-I and these effects were more marked in the cells of the perichondrium than those in the growth plate. However, the most prominent observation was noted in the hypertrophic zone where all treatments containing IGF-I significantly increased (3-fold) the length of this zone, whereas Dex alone had no significant effect. In conclusion, Dex impaired longitudinal growth by inhibiting chondrocyte proliferation, whereas IGF-I stimulated chondrocyte hypertrophy and reversed the growth-inhibitory Dex effects. However, the IGF-I mediated improvement in growth was at the expense of altering the balance between proliferating and hypertrophic chondrocytes within the metatarsal. PMID- 14749360 TI - Sheep exhibit novel variations in the organization of the mammalian type II gonadotropin-releasing hormone receptor gene. AB - Species-specific differences in genes encoding type II GnRH receptor indicate that a functional hepta-helical receptor is produced in monkeys but not in rodents, cows, chimpanzees, or humans. To further investigate the extent of evolutionary differences, we sequenced the type II GnRH receptor gene from wild type Soay sheep. The gene was isolated by long-distance PCR using primers to PEX11beta and RBM8A genes known to flank type II GnRH receptor gene homologues. The gene spans 5.7-kb DNA and was sequenced after shot-gun subcloning. Its novel features include absence of a Pit-1 transcription factor binding site, a premature stop codon (TAG) in exon 1, an in-frame deletion of 51 bp (17 codons) in exon 2, and several nonconservative codon changes. Sheep breed variation in the gene was assessed using genomic DNA in PCR-restriction digest assays for the premature stop codon and in a PCR assay for the deletion. Both characteristics were present in all 15 breeds tested. Receptor gene expression was investigated using poly-A(+) RNA Northern analysis, RT-PCR, and in situ hybridization. An oligonucleotide probe to exon 1 revealed an alternative transcript in testis but not in pituitary gland. No transcripts in testis or pituitary were detectable using an exon 2-3 probe. All tissues examined including multiple brain areas and gonadotrope-enriched cell cultures were negative for type II GnRH receptor in RT PCR. Testis and pituitary sections were negative with exon 1 riboprobes and exon 1 or 2-3 oligonucleotide probes in in situ hybridization. A hepta-helical type II GnRH receptor is therefore not expressed from this sheep gene. PMID- 14749358 TI - Small nuclear RING finger protein expression during gonad development: regulation by gonadotropins and estrogen in the postnatal ovary. AB - Small nuclear RING finger protein (SNURF/RNF4) is a steroid receptor coregulator that is down-regulated in testicular germ cell cancer. In this work, we examined SNURF expression during murine fetal gonad development and postnatal ovarian folliculogenesis by in situ hybridization and immunohistochemical staining. SNURF mRNA was detectable in gonads of both sexes from embryonic 10.5 days post conception onward. SNURF protein localized to gonocytes and somatic Leydig and Sertoli cells of fetal testis and in oogonia and supporting cells of fetal ovary. In murine postnatal ovary, SNURF mRNA and protein were expressed throughout folliculogenesis, peaking in the oocytes of preantral follicles. Lower amounts of SNURF mRNA and protein were also present in granulosa cells of secondary, antral, and preovulatory follicles and in luteal glands. Exposure of immature female mice and rats to gonadotropin from pregnant mare serum and human chorionic gonadotropin did not change dramatically SNURF mRNA levels in ovary. SNURF mRNA expression was increased in ovaries of immature mice treated with diethylstilbestrol, an effect that was blocked by the pure antiestrogen ICI 182,780. SNURF protein was constitutively expressed in oocytes of hypophysectomized rats, and its content was augmented by estradiol in granulosa cells. In granulosa cell culture, SNURF mRNA accumulation was transiently increased by treatment with the LH agonists phorbol myristate and forskolin at 4 h after treatment and at 48 h in differentiated cells expressing markers of the preovulatory phenotype. These results suggest a role for SNURF in fetal germ cell development as well as in oocyte and granulosa cell maturation in an estrogen- and gonadotropin-regulated fashion. PMID- 14749361 TI - Expression of the insulin-like growth factor binding proteins during postnatal development of the murine mammary gland. AB - IGF-I and IGF-II have known roles in postnatal development of the mammary gland. In contrast, the function of the high-affinity IGF binding proteins (IGFBPs) in mammary growth and differentiation is largely unknown. The goal of these studies was to determine the patterns and levels of IGFBP expression during postnatal growth of the murine mammary gland. IGFBP-1 to -5 proteins were detected in mammary tissue by immunoblotting during both pubertal and pregnancy-induced growth; however, the regulation of each IGFBP was distinct through these developmental periods. IGFBP-2 to -5 mRNAs were readily detectable in the developing gland by in situ hybridization analyses but were expressed in distinct cellular sites. IGFBP-3 and -5 mRNAs were expressed in the developing epithelial structures and in isolated stromal cells during ductal growth and alveolar differentiation. In the terminal end buds (TEBs), IGFBP-3 mRNA expression was consistent with its localization in the cap cells, whereas IGFBP-5 was highly expressed in the body cells of the TEB. In contrast, IGFBP-2 and -4 mRNAs were expressed predominantly in stromal cells. IGFBP-2 mRNA was localized to restricted sites in the neck of the TEB and along the ductal structures, whereas IGFBP-4 mRNA was widely expressed in the stroma surrounding the epithelial structures. Protein and mRNA expression for most of the IGFBPs decreased during lactational ages. Levels of IGFBP-2 and -5 protein increased after pup removal during forced involution. Taken together, these data suggest important functions for the family of IGFBPs during postnatal growth and differentiation of the mammary epithelium. PMID- 14749362 TI - SCL assembles a multifactorial complex that determines glycophorin A expression. AB - SCL/TAL1 is a hematopoietic-specific transcription factor of the basic helix-loop helix (bHLH) family that is essential for erythropoiesis. Here we identify the erythroid cell-specific glycophorin A gene (GPA) as a target of SCL in primary hematopoietic cells and show that SCL occupies the GPA locus in vivo. GPA promoter activation is dependent on the assembly of a multifactorial complex containing SCL as well as ubiquitous (E47, Sp1, and Ldb1) and tissue-specific (LMO2 and GATA-1) transcription factors. In addition, our observations suggest functional specialization within this complex, as SCL provides its HLH protein interaction motif, GATA-1 exerts a DNA-tethering function through its binding to a critical GATA element in the GPA promoter, and E47 requires its N-terminal moiety (most likely entailing a transactivation function). Finally, endogenous GPA expression is disrupted in hematopoietic cells through the dominant inhibitory effect of a truncated form of E47 (E47-bHLH) on E-protein activity or of FOG (Friend of GATA) on GATA activity or when LMO2 or Ldb-1 protein levels are decreased. Together, these observations reveal the functional complementarities of transcription factors within the SCL complex and the essential role of SCL as a nucleation factor within a higher-order complex required to activate gene GPA expression. PMID- 14749363 TI - Imbalanced gp130-dependent signaling in macrophages alters macrophage colony stimulating factor responsiveness via regulation of c-fms expression. AB - The mechanisms by which interleukin-6 (IL-6) family cytokines, which utilize the common receptor signaling subunit gp130, influence monocyte/macrophage development remain unclear. Here we have utilized macrophages devoid of either gp130-dependent STAT1/3 (gp130(Delta STAT/Delta STAT)) or extracellular signal regulated kinases 1 and 2 (ERK1/2) mitogen-activated protein (MAP) kinase (gp130(Y757F/Y757F)) activation to assess the individual contribution of each pathway to macrophage formation. While the inhibition by IL-6 of macrophage colony-stimulating factor (M-CSF)-induced colony formation observed in gp130(wt/wt) mice was abolished in gp130(Delta STAT/Delta STAT) mice, inhibition of macrophage colony formation was enhanced in gp130(Y757F/Y757F) mice. In gp130(Delta STAT/Delta STAT) bone marrow-derived macrophages (BMMs), both IL-6- and M-CSF-induced ERK1/2 tyrosine phosphorylation was enhanced. By contrast, tyrosine phosphorylation of ERK1/2 in response to M-CSF was reduced in gp130(Y757F/Y757F) BMMs, and the pattern of ERK1/2 activation in gp130 mutant BMMs correlated with their opposing responsiveness to M-CSF-induced proliferation. When compared to the level of expression in gp130(wt/wt) BMMs, c fms expression was elevated in gp130(Delta STAT/Delta STAT) BMMs but reduced in gp130(Y757F/Y757F) BMMs. Finally, an ERK1/2 inhibitor suppressed M-CSF-induced BMM proliferation, and this result corresponded to a reduction in c-fms expression. Collectively, these results provide a functional and causal correlation between gp130-dependent ERK MAP kinase signaling and c-fms gene activation, a finding that provides a potential mechanism underlying the inhibition of M-CSF-dependent macrophage development by IL-6 family cytokines in mice. PMID- 14749364 TI - Kinase RIP3 is dispensable for normal NF-kappa Bs, signaling by the B-cell and T cell receptors, tumor necrosis factor receptor 1, and Toll-like receptors 2 and 4. AB - RIP3 is a member of the RIP kinase family. It is expressed in the embryo and in multiple adult tissues, including most hemopoietic cell lineages. Several studies have implicated RIP3 in the regulation of apoptosis and NF-kappa B signaling, but whether RIP3 promotes or attenuates activation of the NF-kappa B family of transcription factors has been controversial. We have generated RIP3-deficient mice by gene targeting and find RIP3 to be dispensable for normal mouse development. RIP3-deficient cells showed normal sensitivity to a variety of apoptotic stimuli and were indistinguishable from wild-type cells in their ability to activate NF-kappa B signaling in response to the following: human tumor necrosis factor (TNF), which selectively engages mouse TNF receptor 1; cross-linking of the B- or T-cell antigen receptors; peptidoglycan, which activates Toll-like receptor 2; and lipopolysaccharide (LPS), which stimulates Toll-like receptor 4. Consistent with these observations, RIP3-deficient mice exhibited normal antibody production after immunization with a T-dependent antigen and normal interleukin-1 beta (IL-1 beta), IL-6, and TNF production after LPS treatment. Thus, we can exclude RIP3 as an essential modulator of NF-kappa B signaling downstream of several receptor systems. PMID- 14749365 TI - Studies of the role of the Drosophila scs and scs' insulators in defining boundaries of a chromosome puff. AB - Insulators are DNA elements that establish independent transcriptional domains within eukaryotic genomes. The Drosophila scs and scs' insulators localize near the borders of a structural domain in the polytene chromosomes, known as a puff, produced by transcription of the 87A heat shock protein (hsp) genes. It has been suggested that scs and scs' are boundary elements that delimit this decondensed chromatin domain, reflecting the mechanism by which these sequences act to constrain regulatory interactions. This model was tested using transposons that carried a yellow gene to assess enhancer blocking and an hsp70-lacZ gene to examine the structure of a heat shock puff in the presence and absence of insulators. We found that although scs and scs' blocked enhancer function, these sequences did not prevent the spread of decondensation resulting from hsp70-lacZ transcription. Further analysis of the endogenous 87A locus demonstrated that scs and scs' reside within, not at, the borders of the puff. Taken together, our studies suggest that scs and scs' are not boundary elements that block the propagation of an altered chromatin state associated with puff formation. We propose that these insulators may have a direct role in limiting regulatory interactions in the gene-dense 87A region. PMID- 14749366 TI - Expression of a mutant lamin A that causes Emery-Dreifuss muscular dystrophy inhibits in vitro differentiation of C2C12 myoblasts. AB - Autosomal dominantly inherited missense mutations in lamins A and C cause several tissue-specific diseases, including Emery-Dreifuss muscular dystrophy (EDMD) and Dunnigan-type familial partial lipodystrophy (FPLD). Here we analyze myoblast-to myotube differentiation in C2C12 clones overexpressing lamin A mutated at arginine 453 (R453W), one of the most frequent mutations in EDMD. In contrast with clones expressing wild-type lamin A, these clones differentiate poorly or not at all, do not exit the cell cycle properly, and are extensively committed to apoptosis. These disorders are correlated with low levels of expression of transcription factor myogenin and with the persistence of a large pool of hyperphosphorylated retinoblastoma protein. Since clones mutated at arginine 482 (a site responsible for FPLD) differentiate normally, we conclude that C2C12 clones expressing R453W-mutated lamin A represent a good cellular model to study the pathophysiology of EDMD. Our hypothesis is that lamin A mutated at arginine 453 fails to build a functional scaffold and/or to maintain the chromatin compartmentation required for differentiation of myoblasts into myocytes. PMID- 14749367 TI - Regulation of apoptosis by the Ft1 protein, a new modulator of protein kinase B/Akt. AB - The serine/threonine kinase protein kinase B (PKB)/Akt plays a central role in many cellular processes, including cell growth, glucose metabolism, and apoptosis. However, the identification and validation of novel regulators or effectors is key to future advances in understanding the multiple functions of PKB. Here we report the identification of a novel PKB binding protein, called Ft1, from a cDNA library screen using a green fluorescent protein-based protein fragment complementation assay. We show that the Ft1 protein interacts directly with PKB, enhancing the phosphorylation of both of its regulatory sites by promoting its interaction with the upstream kinase PDK1. Further, the modulation of PKB activity by Ft1 has a strong effect on the apoptosis susceptibility of T lymphocytes treated with glucocorticoids. We demonstrate that this phenomenon occurs via a PDK1/PKB/GSK3/NF-ATc signaling cascade that controls the production of the proapoptotic hormone Fas ligand. The wide distribution of Ft1 in adult tissues suggests that it could be a general regulator of PKB activity in the control of differentiation, proliferation, and apoptosis in many cell types. PMID- 14749368 TI - Protein kinase B/Akt acts via glycogen synthase kinase 3 to regulate recycling of alpha v beta 3 and alpha 5 beta 1 integrins. AB - Protein kinase B (PKB)/Akt is known to promote cell migration, and this may contribute to the enhanced invasiveness of malignant cells. To elucidate potential mechanisms by which PKB/Akt promotes the migration phenotype, we have investigated its role in the endosomal transport and recycling of integrins. Whereas the internalization of alpha v beta 3 and alpha 5 beta 1 integrins and their transport to the recycling compartment were independent of PKB/Akt, the return of these integrins (but not internalized transferrin) to the plasma membrane was regulated by phosphatidylinositol 3-kinases and PKB/Akt. The blockade of integrin recycling and cell spreading on integrin ligands effected by inhibition of PKB/Akt was reversed by inhibition of glycogen synthase kinase 3 (GSK-3). Moreover, expression of nonphosphorylatable active GSK-3 beta mutant GSK 3 beta-A9 suppressed recycling of alpha 5 beta 1 and alpha v beta 3 and reduced cell spreading on ligands for these integrins, indicating that PKB/Akt promotes integrin recycling by phosphorylating and inactivating GSK-3. We propose that the ability of PKB/Akt to act via GSK-3 to promote the recycling of matrix receptors represents a key mechanism whereby integrin function and cell migration can be regulated by growth factors. PMID- 14749369 TI - Activation of H-Ras in the endoplasmic reticulum by the RasGRF family guanine nucleotide exchange factors. AB - Recent findings indicate that in addition to its location in the peripheral plasma membrane, H-Ras is found in endomembranes like the endoplasmic reticulum and the Golgi complex. In these locations H-Ras is functional and can efficiently engage downstream effectors, but little is known about how its activation is regulated in these environments. Here we show that the RasGRF family exchange factors, both endogenous and ectopically expressed, are present in the endoplasmic reticulum but not in the Golgi complex. With the aid of H-Ras constructs specifically tethered to the plasma membrane, endoplasmic reticulum, and Golgi complex, we demonstrate that RasGRF1 and RasGRF2 can activate plasma membrane and reticular, but not Golgi-associated, H-Ras. We also show that RasGRF DH domain is required for the activation of H-Ras in the endoplasmic reticulum but not in the plasma membrane. Furthermore, we demonstrate that RasGRF mediation favors the activation of reticular H-Ras by lysophosphatidic acid treatment whereas plasma membrane H-Ras is made more responsive to stimulation by ionomycin. Overall, our results provide the initial insights into the regulation of H-Ras activation in the endoplasmic reticulum. PMID- 14749370 TI - Role of Gab1 in UV-induced c-Jun NH2-terminal kinase activation and cell apoptosis. AB - Exposure of mammalian cells to UV irradiation leads to activation of the c-Jun NH(2)-terminal protein kinase (JNK) pathway, which is associated with cell apoptosis. However, the molecular mechanism for JNK activation by UV exposure is not fully understood. We show here an essential role of a multisubstrate adapter, Gab1, in this signaling cascade. Gab1-deficient mouse fibroblast cells were defective in induction of JNK activity by UV exposure or heat shock, and this defect was rescued by reintroduction of Gab1 into Gab1(-/-) cells. Consistently, Gab1(-/-) cells displayed reduced caspase 3 induction and apoptotic cell death in response to UV irradiation. Gab1 was constitutively complexed with JNK and became tyrosine phosphorylated in UV-irradiated cells. Genetic and pharmaceutical analyses suggest the involvement of c-Met and the Src family tyrosine kinases in mediating UV-induced Gab1 phosphorylation as well as JNK activation. In aggregate, these observations identify a new function of Gab1 in the response of mammalian cells to UV light. PMID- 14749371 TI - Invasion of normal human fibroblasts induced by v-Fos is independent of proliferation, immortalization, and the tumor suppressors p16INK4a and p53. AB - Invasion is generally perceived to be a late event during the progression of human cancer, but to date there are no consistent reports of alterations specifically associated with malignant conversion. We provide evidence that the v Fos oncogene induces changes in gene expression that render noninvasive normal human diploid fibroblasts highly invasive, without inducing changes in growth factor requirements or anchorage dependence for proliferation. Furthermore, v-Fos stimulated invasion is independent of the pRb/p16(INK4a) and p53 tumor suppressor pathways and telomerase. We have performed microarray analysis using Affymetrix GeneChips, and the gene expression profile of v-Fos transformed cells supports its role in the regulation of invasion, independent from proliferation. We also demonstrate that invasion, but not proliferation, is dependent on the activity of the up-regulated epidermal growth factor receptor. Taken together, these results indicate that AP-1-directed invasion could precede deregulated proliferation during tumorigenesis and that sustained activation of AP-1 could be the epigenetic event required for conversion of a benign tumor into a malignant one, thereby explaining why many malignant human tumors present without an obvious premalignant hyperproliferative dysplastic lesion. PMID- 14749372 TI - Mnt loss triggers Myc transcription targets, proliferation, apoptosis, and transformation. AB - Myc oncoproteins are overexpressed in most cancers and are sufficient to accelerate cell proliferation and provoke transformation. However, in normal cells Myc also triggers apoptosis. All of the effects of Myc require its function as a transcription factor that dimerizes with Max. This complex induces genes containing CACGTG E-boxes, such as Ornithine decarboxylase (Odc), which harbors two of these elements. Here we report that in quiescent cells the Odc E-boxes are occupied by Max and Mnt, a putative Myc antagonist, and that this complex is displaced by Myc-Max complexes in proliferating cells. Knockdown of Mnt expression by stable retroviral RNA interference triggers many targets typical of the "Myc" response and provokes accelerated proliferation and apoptosis. Strikingly, these effects of Mnt knockdown are even manifest in cells lacking c myc. Moreover, Mnt knockdown is sufficient to transform primary fibroblasts in conjunction with Ras. Therefore, Mnt behaves as a tumor suppressor. These findings support a model where Mnt represses Myc target genes and Myc functions as an oncogene by relieving Mnt-mediated repression. PMID- 14749373 TI - Proapoptotic BH3-only Bcl-2 family member Bik/Blk/Nbk is expressed in hemopoietic and endothelial cells but is redundant for their programmed death. AB - The BH3-only members of the Bcl-2 protein family are essential for initiation of programmed cell death and stress-induced apoptosis. We have determined the expression pattern in mice of the BH3-only protein Bik, also called Blk or Nbk, and examined its physiological function by gene targeting. We found that Bik is expressed widely in the hematopoietic compartment and in endothelial cells of the venous but not arterial lineages. Nevertheless, its loss did not increase the numbers of such cells in mice or protect hematopoietic cells in vitro from apoptosis induced by cytokine withdrawal or diverse other cytotoxic stimuli. Moreover, whereas loss of the BH3-only protein Bim rescued mice lacking the prosurvival protein Bcl-2 from fatal polycystic kidney disease and lymphopenia, loss of Bik did not. These results indicate that any function of Bik in programmed cell death and stress-induced apoptosis must overlap that of other BH3 only proteins. PMID- 14749374 TI - Negative control of the Myc protein by the stress-responsive kinase Pak2. AB - Pak2 is a serine/threonine kinase that participates in the cellular response to stress. Among the potential substrates for Pak2 is the protein Myc, encoded by the proto-oncogene MYC. Here we demonstrate that Pak2 phosphorylates Myc at three sites (T358, S373, and T400) and affects Myc functions both in vitro and in vivo. Phosphorylation at all three residues reduces the binding of Myc to DNA, either by blocking the requisite dimerization with Max (through phosphorylation at S373 and T400) or by interfering directly with binding to DNA (through phosphorylation at T358). Phosphorylation by Pak2 inhibits the ability of Myc to activate transcription, to sustain cellular proliferation, to transform NIH 3T3 cells in culture, and to elicit apoptosis on serum withdrawal. These results indicate that Pak2 is a negative regulator of Myc, suggest that inhibition of Myc plays a role in the cellular response to stress, and raise the possibility that Pak2 may be the product of a tumor suppressor gene. PMID- 14749375 TI - Functional interaction between poly(ADP-Ribose) polymerase 2 (PARP-2) and TRF2: PARP activity negatively regulates TRF2. AB - The DNA damage-dependent poly(ADP-ribose) polymerase-2 (PARP-2) is, together with PARP-1, an active player of the base excision repair process, thus defining its key role in genome surveillance and protection. Telomeres are specialized DNA protein structures that protect chromosome ends from being recognized and processed as DNA strand breaks. In mammals, telomere protection depends on the T(2)AG(3) repeat binding protein TRF2, which has been shown to remodel telomeres into large duplex loops (t-loops). In this work we show that PARP-2 physically binds to TRF2 with high affinity. The association of both proteins requires the N terminal domain of PARP-2 and the myb domain of TRF2. Both partners colocalize at promyelocytic leukemia bodies in immortalized telomerase-negative cells. In addition, our data show that PARP activity regulates the DNA binding activity of TRF2 via both a covalent heteromodification of the dimerization domain of TRF2 and a noncovalent binding of poly(ADP-ribose) to the myb domain of TRF2. PARP-2( /-) primary cells show normal telomere length as well as normal telomerase activity compared to wild-type cells but display a spontaneously increased frequency of chromosome and chromatid breaks and of ends lacking detectable T(2)AG(3) repeats. Altogether, these results suggest a functional role of PARP-2 activity in the maintenance of telomere integrity. PMID- 14749376 TI - Myodegeneration in EDA-A2 transgenic mice is prevented by XEDAR deficiency. AB - EDA-A1 and EDA-A2 are members of the tumor necrosis factor family of ligands. The products of alternative splicing of the ectodysplasin (EDA) gene, EDA-A1 and EDA A2 differ by an insertion of two amino acids and bind to distinct receptors. The longer isoform, EDA-A1, binds to EDAR and plays an important role in sweat gland, hair, and tooth development; mutations in EDA, EDAR, or the downstream adaptor EDARADD cause hypohidrotic ectodermal dysplasia. EDA-A2 engages the receptor XEDAR, but its role in the whole organism is less clear. We have generated XEDAR deficient mice by gene targeting and transgenic mice expressing secreted forms of EDA-A1 or EDA-A2 downstream of the skeletal muscle-specific myosin light-chain 2 or skin-specific keratin 5 promoter. Mice lacking XEDAR were indistinguishable from their wild-type littermates, but EDA-A2 transgenic mice exhibited multifocal myodegeneration. This phenotype was not observed in the absence of XEDAR. Skeletal muscle in EDA-A1 transgenic mice was unaffected, but their sebaceous glands were hypertrophied and hyperplastic, consistent with a role for EDA-A1 in the development of these structures. These data indicate that XEDAR-transduced signals are dispensable for development of ectoderm-derived organs but might play a role in skeletal muscle homeostasis. PMID- 14749377 TI - Cdc6 chromatin affinity is unaffected by serine-54 phosphorylation, S-phase progression, and overexpression of cyclin A. AB - Ectopically expressed Cdc6 is translocated from the nucleus during S phase in a cyclin A-Cdk2-dependent process, suggesting that reinitiation of DNA replication is prevented by removal of phosphorylated Cdc6 from chromatin after origin firing. However, whether endogenous Cdc6 translocates during S phase remains controversial. To resolve the questions regarding regulation of endogenous Cdc6, we cloned the cDNA encoding the Chinese hamster Cdc6 homolog and specifically focused on analyzing the localizations and chromatin affinities of endogenous and exogenous proteins during S phase and following overexpression of cyclin A. In agreement with other reports, ectopically expressed Cdc6 translocates from the nucleus during S phase and in response to overexpressed cyclin A. In contrast, using a combination of biochemical and immunohistochemical assays, we show convincingly that endogenous Cdc6 remains nuclear and chromatin bound throughout the entire S period, while Mcm5 loses chromatin affinity during S phase. Overexpression of cyclin A is unable to alter the nuclear localization of Cdc6. Furthermore, using a phosphospecific antibody we show that phosphoserine-54 Cdc6 maintains a high affinity for chromatin during the S period. Considering recent in vitro studies, these data are consistent with a proposed model in which Cdc6 is serine-54 phosphorylated during S phase and functions as a chromatin-bound signal that prevents reformation of prereplication complexes. PMID- 14749378 TI - Actin cytoskeleton regulates calcium dynamics and NFAT nuclear duration. AB - T-cell activation by antigen-presenting cells is accompanied by actin polymerization, T-cell receptor (TCR) capping, and formation of the immunological synapse. However, whether actin-dependent events are required for T-cell function is poorly understood. Herein, we provide evidence for an unexpected negative regulatory role of the actin cytoskeleton on TCR-induced cytokine production. Disruption of actin polymerization resulted in prolonged intracellular calcium elevation in response to anti-CD3, thapsigargin, or phorbol myristate acetate plus ionomycin, leading to persistent NFAT (nuclear factor of activated T cells) nuclear duration. These events were dominant, as the net effect of actin blockade was augmented interleukin 2 promoter activity. Increased surface expression of the plasma membrane Ca(2+) ATPase was observed upon stimulation, which was inhibited by cytochalasin D, suggesting that actin polymerization contributes to calcium export. Our results imply a novel role for the actin cytoskeleton in modulating the duration of Ca(2+)-NFAT signaling and indicate that actin dynamics regulate features of T-cell activation downstream of receptor clustering. PMID- 14749379 TI - Dnmt1 expression in pre- and postimplantation embryogenesis and the maintenance of IAP silencing. AB - The methylation of intracisternal A-type particle (IAP) sequences is maintained during mouse embryogenesis. Methylation suppresses IAP expression and the potential for mutagenesis by retrotransposition, but it is not clear how methylation of these elements is maintained during the embryonic stages when the bulk of the genome is being demethylated. It has been suggested that the high levels of DNA methyltransferase-1 (Dnmt1) present during cleavage could be important for keeping IAPs methylated. To test this hypothesis, we combined mutant alleles of Dnmt1 with an agouti allele (A(iapy)), which provided a coat color readout for the methylation status of the IAP insertion in the agouti locus. We found that reduction in Dnmt1 levels directly impacted methylation at this locus, leading to stable transcriptional activation of the agouti gene in the adult. Specifically, the short maternal Dnmt1 protein was important in maintaining methylation at the A(iapy) locus in cleavage embryos, whereas the longer Dnmt1 isoform found in somatic cells was important in maintaining IAP methylation during the postimplantation stage. These results underscore the importance of maintaining proper maintenance of methylation patterns during gestation and suggest that interference with this process may stably affect gene expression patterns in the adult and may have profound phenotypic consequences. PMID- 14749380 TI - Capn5 is expressed in a subset of T cells and is dispensable for development. AB - The Capn5 gene was inactivated by homologous recombination in ES cells that subsequently colonized the germ line of mice. The targeted mutation integrated a lacZ expression cassette into the Capn5 gene, allowing the expression of Capn5 mRNA to be examined in detail in heterozygous animals. Expression was observed in embryonic and newborn thymuses, in various epithelial tissues, and in tissues of the central nervous system. In the thymus, Capn5 was expressed mainly in relatively immature CD25(+) embryonic thymocytes. Despite the numerous expression sites of Capn5, the majority of Capn5-null mice were viable and fertile and appeared healthy. Histopathological analysis did not reveal any differences between Capn5-null and wild-type mice. There were no defects in the major T- or B cell populations in the thymus, spleen, bone marrow, or peritoneum, nor did apoptosis appear abnormal in Capn5-null T cells. There was no evidence for the development of autoimmune disease in Capn5-null animals. However, a small proportion of homozygous null offspring from heterozygous matings were runted and most often did not survive to adulthood. PMID- 14749381 TI - Mouse strains with an active H2-Ea meiotic recombination hot spot exhibit increased levels of H2-Ea-specific DNA breaks in testicular germ cells. AB - We devised a sensitive method for the site-specific detection of rare meiotic DNA strand breaks in germ cell-enriched testicular cell populations from mice that possess or lack an active recombination hot spot at the H2-Ea gene. Using germ cells from adult animals, we found an excellent correlation between the frequency of DNA breaks in the 418-bp H2-Ea hot spot and crossover activity. The temporal appearance of DNA breaks was also studied in 7- to 18-day-old mice with an active hot spot during the first waves of spermatogenesis. The number of DNA breaks detected rose as leptotene and zygotene spermatocytes populate the testis with a peak at day 14 postpartum, when leptotene, zygotene, and early pachytene spermatocytes are the most common meiotic prophase I cell types. The number of DNA breaks drops precipitously 1 day later, when middle to late pachytene spermatocytes become the dominant subtype. The recombination-related breaks in the hot spot likely reflect SPO11-induced double-strand breaks and/or recombination intermediates containing free 3' hydroxyl groups. PMID- 14749382 TI - Placental failure and impaired vasculogenesis result in embryonic lethality for neuropathy target esterase-deficient mice. AB - Age-dependent neurodegeneration resulting from widespread apoptosis of neurons and glia characterize the Drosophila Swiss Cheese (SWS) mutant. Neuropathy target esterase (NTE), the vertebrate homologue of SWS, reacts with organophosphates which initiate a syndrome of axonal degeneration. NTE is expressed in neurons and a variety of nonneuronal cell types in adults and fetal mice. To investigate the physiological functions of NTE, we inactivated its gene by targeted mutagenesis in embryonic stem cells. Heterozygous NTE(+/-) mice displayed a 50% reduction in NTE activity but underwent normal organ development. Complete inactivation of the NTE gene resulted in embryonic lethality, which became evident after gastrulation at embryonic day 9 postcoitum (E9). As early as E7.5, mutant embryos revealed growth retardation which did not reflect impaired cell proliferation but rather resulted from failed placental development; as a consequence, massive apoptosis within the developing embryo preceded its resorption. Histological analysis indicated that NTE is essential for the formation of the labyrinth layer and survival and differentiation of secondary giant cells. Additionally, impairment of vasculogenesis in the yolk sacs and embryos of null mutant conceptuses suggested that NTE is also required for normal blood vessel development. PMID- 14749383 TI - Ogt-dependent X-chromosome-linked protein glycosylation is a requisite modification in somatic cell function and embryo viability. AB - The Ogt gene encodes a glycosyltransferase that links N-acetylglucosamine to serine and threonine residues (O-GlcNAc) on nuclear and cytosolic proteins. Efforts to study a mammalian model of Ogt deficiency have been hindered by the requirement for this X-linked gene in embryonic stem cell viability, necessitating the use of conditional mutagenesis in vivo. We have extended these observations by segregating Ogt mutation to distinct somatic cell types, including neurons, thymocytes, and fibroblasts, the latter by an approach developed for inducible Ogt mutagenesis. We show that Ogt mutation results in the loss of O-GlcNAc and causes T-cell apoptosis, neuronal tau hyperphosphorylation, and fibroblast growth arrest with altered expression of c-Fos, c-Jun, c-Myc, Sp1, and p27. We further segregated the mutant Ogt allele to parental gametes by oocyte- and spermatid-specific Cre-loxP mutagenesis. By this we established an in vivo genetic approach that supports the ontogeny of female heterozygotes bearing mutant X-linked genes required during embryogenesis. Successful production and characterization of such female heterozygotes further indicates that mammalian cells commonly require a functional Ogt allele. We find that O-GlcNAc modulates protein phosphorylation and expression among essential and conserved cell signaling pathways. PMID- 14749384 TI - Matrilin-3 is dispensable for mouse skeletal growth and development. AB - Matrilin-3 belongs to the matrilin family of extracellular matrix (ECM) proteins and is primarily expressed in cartilage. Mutations in the gene encoding human matrilin-3 (MATN-3) lead to autosomal dominant skeletal disorders, such as multiple epiphyseal dysplasia (MED), which is characterized by short stature and early-onset osteoarthritis, and bilateral hereditary microepiphyseal dysplasia, a variant form of MED characterized by pain in the hip and knee joints. To assess the function of matrilin-3 during skeletal development, we have generated Matn-3 null mice. Homozygous mutant mice appear normal, are fertile, and show no obvious skeletal malformations. Histological and ultrastructural analyses reveal endochondral bone formation indistinguishable from that of wild-type animals. Northern blot, immunohistochemical, and biochemical analyses indicated no compensatory upregulation of any other member of the matrilin family. Altogether, our findings suggest functional redundancy among matrilins and demonstrate that the phenotypes of MED disorders are not caused by the absence of matrilin-3 in cartilage ECM. PMID- 14749385 TI - Recycling of the U12-type spliceosome requires p110, a component of the U6atac snRNP. AB - U12-dependent introns are spliced by the so-called minor spliceosome, requiring the U11, U12, and U4atac/U6atac snRNPs in addition to the U5 snRNP. We have recently identified U6-p110 (SART3) as a novel human recycling factor that is related to the yeast splicing factor Prp24. U6-p110 transiently associates with the U6 and U4/U6 snRNPs during the spliceosome cycle, regenerating functional U4/U6 snRNPs from singular U4 and U6 snRNPs. Here we investigated the involvement of U6-p110 in recycling of the U4atac/U6atac snRNP. In contrast to the major U6 and U4/U6 snRNPs, p110 is primarily associated with the U6atac snRNP but is almost undetectable in the U4atac/U6atac snRNP. Since p110 does not occur in U5 snRNA-containing complexes, it appears to be transiently associated with U6atac during the cycle of the minor spliceosome. The p110 binding site was mapped to U6 nucleotides 38 to 57 and U6atac nucleotides 10 to 30, which are highly conserved between these two functionally related snRNAs. With a U12-dependent in vitro splicing system, we demonstrate that p110 is required for recycling of the U4atac/U6atac snRNP. PMID- 14749386 TI - RNA polymerase II (Pol II)-TFIIF and Pol II-mediator complexes: the major stable Pol II complexes and their activity in transcription initiation and reinitiation. AB - Protein purification and depletion studies were used to determine the major stable forms of RNA polymerase II (Pol II) complexes found in Saccharomyces cerevisiae nuclear extracts. About 50% of Pol II is found associated with the general transcription factor TFIIF (Pol II-TFIIF), and about 20% of Pol II is associated with Mediator (Pol-Med). No Pol II-Med-TFIIF complex was observed. The activity of Pol II and the purified Pol II complexes in transcription initiation and reinitiation was investigated by supplementing extracts depleted of either total Pol II or total TFIIF with purified Pol II or the Pol II complexes. We found that all three forms of Pol II can complement Pol II-depleted extracts for transcription initiation, but Pol II-TFIIF has the highest specific activity. Similarly, Pol II-TFIIF has a much higher specific activity than TFIIF for complementation of TFIIF transcription activity. Although the Pol II-TFIIF and Pol II-Med complexes were stable when purified, we found these complexes were dynamic in extracts under transcription conditions, with a single polymerase capable of exchanging bound Mediator and TFIIF. Using a purified system to examine transcription reinitiation, we found that Pol II-TFIIF was active in promoting multiple rounds of transcription while Pol II-Med was nearly inactive. These results suggest that both the Pol II-Med and Pol II-TFIIF complexes can be recruited for transcription initiation but that only the Pol II-TFIIF complex is competent for transcription reinitiation. PMID- 14749387 TI - Two cyclin-dependent kinases promote RNA polymerase II transcription and formation of the scaffold complex. AB - Three cyclin-dependent kinases, CDK7, -8, and -9, are specifically involved in transcription by RNA polymerase II (Pol II) and target the Pol II C-terminal domain (CTD). The role of CDK7 and CDK8 kinase activity in transcription has been unclear, with CDK7 shown to have variable effects on transcription and CDK8 suggested to repress transcription and/or to target other gene-specific factors. Using a chemical genetics approach, the Saccharomyces cerevisiae homologs of these kinases, Kin28 and Srb10, were engineered to respond to a specific inhibitor and the inhibitor was used to test the role of these kinases in transcription in vivo and in vitro. In vitro, these kinases can both promote transcription, with up to 70% of transcription abolished when both kinases are inhibited together. Similarly, in vivo inhibition of both kinases together gives the strongest decrease in transcription, as measured by chromatin immunoprecipitation of Pol II. Kin28 and Srb10 also have overlapping roles in promoting ATP-dependent dissociation of the preinitiation complex (PIC) into the Scaffold complex. Using the engineered kinases and an ATP analog, specific kinase substrates within the PIC were identified. In addition to the previously known substrate, the Pol II CTD, it was found that Kin28 phosphorylates two subunits of Mediator and Srb10 targets two subunits of TFIID for phosphorylation. PMID- 14749388 TI - Human CNK1 acts as a scaffold protein, linking Rho and Ras signal transduction pathways. AB - Rho family GTPases act as molecular switches to control a variety of cellular responses, including cytoskeletal rearrangements, changes in gene expression, and cell transformation. In the active, GTP-bound state, Rho interacts with an ever growing number of effector molecules, which promote distinct biochemical pathways. Here, we describe the isolation of hCNK1, the human homologue of Drosophila connector enhancer of ksr, as an effector for Rho. hCNK1 contains several protein-protein interaction domains, and Rho interacts with one of these, the PH domain, in a GTP-dependent manner. A mutant hCNK1, which is unable to bind to Rho, or depletion of endogenous hCNK1 by using RNA interference inhibits Rho induced gene expression via serum response factor but has no apparent effect on Rho-induced stress fiber formation, suggesting that it acts as a specific effector for transcriptional, but not cytoskeletal, activation pathways. Finally, hCNK1 associates with Rhophilin and RalGDS, Rho and Ras effector molecules, respectively, suggesting that it acts as a scaffold protein to mediate cross talk between the two pathways. PMID- 14749389 TI - p66SHC promotes apoptosis and antagonizes mitogenic signaling in T cells. AB - Of the three Shc isoforms, p66Shc is responsible for fine-tuning p52/p46Shc signaling to Ras and has been implicated in apoptotic responses to oxidative stress. Here we show that human peripheral blood lymphocytes and mouse thymocytes and splenic T cells acquire the capacity to express p66Shc in response to apoptogenic stimulation. Using a panel of T-cell transfectants and p66Shc(-/-) T cells, we show that p66Shc expression results in increased susceptibility to apoptogenic stimuli, which depends on Ser36 phosphorylation and correlates with an altered balance in apoptosis-regulating gene expression. Furthermore, p66Shc blunts mitogenic responses to T-cell receptor engagement, at least in part by transdominant inhibition of p52Shc signaling to Ras/mitogen-activated protein kinases, in an S36-dependent manner. The data highlight a novel interplay between p66Shc and p52Shc in the control of T-cell fate. PMID- 14749390 TI - TRAM2 protein interacts with endoplasmic reticulum Ca2+ pump Serca2b and is necessary for collagen type I synthesis. AB - Cotranslational insertion of type I collagen chains into the lumen of the endoplasmic reticulum (ER) and their subsequent folding into a heterotrimeric helix is a complex process which requires coordinated action of the translation machinery, components of translocons, molecular chaperones, and modifying enzymes. Here we describe a role for the protein TRAM2 in collagen type I expression in hepatic stellate cells (HSCs) and fibroblasts. Activated HSCs are collagen-producing cells in the fibrotic liver. Quiescent HSCs produce trace amounts of type I collagen, while upon activation collagen synthesis increases 50 to 70-fold. Likewise, expression of TRAM2 dramatically increases in activated HSCs. TRAM2 shares 53% amino acid identity with the protein TRAM, which is a component of the translocon. However, TRAM2 has a C terminus with only a 15% identity. The C-terminal part of TRAM2 interacts with the Ca(2+) pump of the ER, SERCA2b, as demonstrated in a Saccharomyces cerevisiae two-hybrid screen and by immunoprecipitations in human cells. TRAM2 also coprecipitates with anticollagen antibody, suggesting that these two proteins interact. Deletion of the C-terminal part of TRAM2 inhibits type I collagen synthesis during activation of HSCs. The pharmacological inhibitor of SERCA2b, thapsigargin, has a similar effect. Depletion of ER Ca(2+) with thapsigargin results in inhibition of triple helical collagen folding and increased intracellular degradation. We propose that TRAM2, as a part of the translocon, is required for the biosynthesis of type I collagen by coupling the activity of SERCA2b with the activity of the translocon. This coupling may increase the local Ca(2+) concentration at the site of collagen synthesis, and a high Ca(2+) concentration may be necessary for the function of molecular chaperones involved in collagen folding. PMID- 14749391 TI - U17/snR30 is a ubiquitous snoRNA with two conserved sequence motifs essential for 18S rRNA production. AB - Saccharomyces cerevisiae snR30 is an essential box H/ACA small nucleolar RNA (snoRNA) required for the processing of 18S rRNA. Here, we show that the previously characterized human, reptilian, amphibian, and fish U17 snoRNAs represent the vertebrate homologues of yeast snR30. We also demonstrate that U17/snR30 is present in the fission yeast Schizosaccharomyces pombe and the unicellular ciliated protozoan Tetrahymena thermophila. Evolutionary comparison revealed that the 3'-terminal hairpins of U17/snR30 snoRNAs contain two highly conserved sequence motifs, the m1 (AUAUUCCUA) and m2 (AAACCAU) elements. Mutation analysis of yeast snR30 demonstrated that the m1 and m2 elements are essential for early cleavages of the 35S pre-rRNA and, consequently, for the production of mature 18S rRNA. The m1 and m2 motifs occupy the opposite strands of an internal loop structure, and they are located invariantly 7 nucleotides upstream from the ACA box of U17/snR30 snoRNAs. U17/snR30 is the first identified box H/ACA snoRNA that possesses an evolutionarily conserved role in the nucleolytic processing of eukaryotic pre-rRNA. PMID- 14749392 TI - Cleavage of poly(A)-binding protein by poliovirus 3C protease inhibits host cell translation: a novel mechanism for host translation shutoff. AB - Cleavage of eukaryotic translation initiation factor 4GI (eIF4GI) by viral 2A protease (2Apro) has been proposed to cause severe translation inhibition in poliovirus-infected cells. However, infections containing 1 mM guanidine-HCl result in eIF4GI cleavage but only partial translation shutoff, indicating eIF4GI cleavage is insufficient for drastic translation inhibition. Viral 3C protease (3Cpro) cleaves poly(A)-binding protein (PABP) and removes the C-terminal domain (CTD) that interacts with several translation factors. In HeLa cell translation extracts that exhibit cap-poly(A) synergy, partial cleavage of PABP by 3Cpro inhibited translation of endogenous mRNAs and reporter RNA as effectively as complete cleavage of eIF4GI and eIF4GII by 2Apro. 3Cpro-mediated translation inhibition was poly(A) dependent, and addition of PABP to extracts restored translation. Expression of 3Cpro in HeLa cells resulted in partial PABP cleavage and similar inhibition of translation. PABP cleavage did not affect eIF4GI-PABP interactions, and the results of kinetics experiments suggest that 3Cpro might inhibit late steps in translation or ribosome recycling. The data illustrate the importance of the CTD of PABP in poly(A)-dependent translation in mammalian cells. We propose that enteroviruses use a dual strategy for host translation shutoff, requiring cleavage of PABP by 3Cpro and of eIF4G by 2Apro. PMID- 14749393 TI - Recruitment of the nucleolar remodeling complex NoRC establishes ribosomal DNA silencing in chromatin. AB - The rRNA gene cluster consists of multiple transcription units. Half of these are active, while the other half are transcriptionally inactive. Previously, in vivo studies have demonstrated that silencing of ribosomal DNA (rDNA) is mediated by the chromatin remodeling NoRC (nucleolar remodeling complex). To explore the mechanisms underlying NoRC-directed silencing of rDNA transcription, we investigated the effect of recombinant NoRC on RNA polymerase I transcription on reconstituted chromatin templates. We show that NoRC interacts with the transcription terminator factor (TTF-I), and this interaction is required both for the binding of TTF-I to its promoter-proximal target site and for the recruitment of NoRC to the promoter. After association with the rDNA promoter, NoRC alters the position of the promoter-bound nucleosome, thereby repressing RNA polymerase I transcription. This NoRC-directed rDNA repression requires the N terminus of histone H4. Repression is effective before preinitiation complex formation and as such is unable to exert an effect upon activated rDNA genes. Furthermore, the early steps of rDNA repression do not depend on DNA and histone modifications. These results reveal an important role for TTF-I in recruiting NoRC to rDNA and an active role for NoRC in the establishment of rDNA silencing. PMID- 14749394 TI - Enhancer I predominance in hepatitis B virus gene expression. AB - Previous studies of human hepatitis B virus (HBV) transcription revealed the requirement of two enhancer elements. Enhancer I (EnhI) is located upstream of the X promoter and is targeted by multiple activators, including basic leucine zipper proteins, and enhancer II (EnhII) is located upstream to the PreCore promoter and is targeted mainly by nuclear receptors (NRs). The mode of interplay between these enhancers and their unique contributions in regulating HBV transcription remained obscure. By using time course analysis we revealed that the HBV transcripts are categorized into early and late groups. Chang (CCL-13) cells are impaired in expression of the late transcripts. This could be corrected by overexpressing EnhII activators, such as hepatocyte nuclear factor 4 alpha, the retinoid X receptor alpha, and the peroxisome proliferator-activated receptor alpha, suggesting that in Chang cells EnhI but not EnhII is active. Replacing the 5'-end EnhI sequence with a synthetic Gal4 response (UAS) DNA fragment ceased the production of the early transcripts. Under this condition NR overexpression poorly activated EnhII. However, activation of the UAS by Gal4-p53 restored both the expression of the early transcripts and the EnhII response to NRs. Thus, a functional EnhI is required for activation of EnhII. We found a major difference between Gal4-p53 and Gal4-VP16 behavior. Gal4-p53 activated the early transcripts, while Gal4-VP16 inhibited the early transcripts but activated the late transcripts. These findings indicate that the composition of the EnhI binding proteins may play a role in early to late switching. Our data provides strong evidence for the role of EnhI in regulating global and temporal HBV gene expression. PMID- 14749395 TI - Mutant MyoD lacking Cdc2 phosphorylation sites delays M-phase entry. AB - The transcription factors MyoD and Myf-5 control myoblast identity and differentiation. MyoD and Myf-5 manifest opposite cell cycle-specific expression patterns. Here, we provide evidence that MyoD plays a pivotal role at the G(2)/M transition by controlling the expression of p21(Waf1/Cip1) (p21), which is believed to regulate cyclin B-Cdc2 kinase activity in G(2). In growing myoblasts, MyoD reaccumulates during G(2) concomitantly with p21 before entry into mitosis; MyoD is phosphorylated on Ser5 and Ser200 by cyclin B-Cdc2, resulting in a decrease of its stability and down-regulation of both MyoD and p21. Inducible expression of a nonphosphorylable MyoD A5/A200 enhances the MyoD interaction with the coactivator P/CAF, thereby stimulating the transcriptional activation of a luciferase reporter gene placed under the control of the p21 promoter. MyoD A5/A200 causes sustained p21 expression, which inhibits cyclin B-Cdc2 kinase activity in G(2) and delays M-phase entry. This G(2) arrest is not observed in p21(-/-) cells. These results show that in cycling cells MyoD functions as a transcriptional activator of p21 and that MyoD phosphorylation is required for G(2)/M transition. PMID- 14749396 TI - Food advertising and childhood obesity. PMID- 14749397 TI - Patient empowerment or the emperor's new clothes. PMID- 14749398 TI - Perinatal risk factors for neonaticide and infant homicide: can we identify those at risk? PMID- 14749399 TI - Chronic non-cancer pain and opioid dependence. PMID- 14749400 TI - Statins for primary prevention: strategic options to save lives and money. PMID- 14749401 TI - Swallowed partial dentures. AB - Swallowed or inhaled partial dentures can present a diagnostic challenge. Three new cases are described, one of them near-fatal because of vascular erosion and haemorrhage. The published work points to the importance of good design and proper maintenance. The key to early recognition is awareness of the hazard by denture-wearers, carers and clinicians. PMID- 14749402 TI - Swimming after meningitis. PMID- 14749403 TI - Spontaneous and permanent resolution of cystic adventitial disease of the popliteal artery. PMID- 14749404 TI - A dog bite to the eyelid. PMID- 14749405 TI - Mucous patches and arthralgia. PMID- 14749406 TI - Aortic stenosis and gastrointestinal bleeding. PMID- 14749407 TI - Recurrent pancreatitis: not just alcohol, gallstones and scorpion venom. PMID- 14749408 TI - A leech in the large bowel. PMID- 14749409 TI - Nymphaea cults in ancient Egypt and the New World: a lesson in empirical pharmacology. PMID- 14749410 TI - Shell shock, Gordon Holmes and the Great War. PMID- 14749411 TI - Castell coch. PMID- 14749412 TI - Art and medicine 2: Pethidine by Dr Matthew Clark. PMID- 14749413 TI - Legal considerations of clinical guidelines. PMID- 14749414 TI - Legal considerations of clinical guidelines. PMID- 14749415 TI - Vocational rehabilitation. PMID- 14749416 TI - Conversion disorder in childhood. PMID- 14749417 TI - Philoctetes. PMID- 14749418 TI - Atypical manifestations of gastrooesophageal reflux. PMID- 14749421 TI - Prefrontal cortical-ventral striatal interactions involved in affective modulation of attentional performance: implications for corticostriatal circuit function. AB - Anatomically segregated systems linking the frontal cortex and the striatum are involved in various aspects of cognitive, affective, and motor processing. In this study, we examined the effects of combined unilateral lesions of the medial prefrontal cortex (mPFC) and the core subregion of the nucleus accumbens (AcbC) in opposite hemispheres (disconnection) on a continuous performance, visual attention test [five-choice serial reaction-time task (5CSRTT)]. The disconnection lesion produced a set of specific changes in performance of the 5CSRTT, resembling changes that followed bilateral AcbC lesions while, in addition, comprising a subset of the behavioral changes after bilateral mPFC lesions previously reported using the same task. Specifically, both mPFC/AcbC disconnection and bilateral AcbC lesions markedly affected aspects of response control related to affective feedback, as indexed by perseverative responding in the 5CSRTT. These effects were comparable, although not identical, to those in animals with either bilateral AcbC or mPFC/AcbC disconnection lesions. The mPFC/AcbC disconnection resulted in a behavioral profile largely distinct from that produced by disconnection of a similar circuit described previously, between the mPFC and the dorsomedial striatum, which were shown to form a functional network underlying aspects of visual attention and attention to action. This distinction provides an insight into the functional specialization of corticostriatal circuits in similar behavioral contexts. PMID- 14749422 TI - A circadian rhythm in the expression of PERIOD2 protein reveals a novel SCN controlled oscillator in the oval nucleus of the bed nucleus of the stria terminalis. AB - Circadian rhythms in mammals are regulated not only globally by the master clock in the suprachiasmatic nucleus (SCN), but also locally by widely distributed populations of clock cells in the brain and periphery that control tissue specific rhythmic outputs. Here we show that the oval nucleus of the bed nucleus of the stria terminalis (BNST-OV) exhibits a robust circadian rhythm in expression of the Period2 (PER2) clock protein. PER2 expression is rhythmic in the BNST-OV in rats housed under a light/dark cycle or in constant darkness, in blind rats, and in mice, and is in perfect synchrony with the PER2 rhythm of the SCN. Constant light or bilateral SCN lesions abolish the rhythm of PER2 in the BNST-OV. Large abrupt shifts in the light schedule transiently uncouple the BNST OV rhythm from that of the SCN. Re-entrainment of the PER2 rhythm is faster in the SCN than in the BNST-OV, and it is faster after a delay than an advance shift. Bilateral adrenalectomy blunts the PER2 rhythm in the BNST-OV. Thus, the BNST-OV contains circadian clock cells that normally oscillate in synchrony with the SCN, but these cells appear to require both input from the SCN and circulating glucocorticoids to maintain their circadian oscillation. Taken together with what is known about the functional organization of the connections of the BNST-OV with systems of the brain involved in stress and motivational processes, these findings place BNST-OV oscillators in a position to influence specific physiological and behavioral rhythms downstream from the SCN clock. PMID- 14749423 TI - CRE-mediated transcription is increased in Huntington's disease transgenic mice. AB - Disruption of cAMP response element (CRE)-dependent transcription has been hypothesized to contribute to neuronal death and dysfunction in Huntington's disease (HD) and other polyglutamine repeat disorders. Whether dysregulation of CRE-dependent transcription actually occurs in vivo in response to expression of expanded polyglutamine repeats has not been tested. We directly tested whether CRE-dependent transcription is affected in vivo by cross breeding a transgenic mouse model of HD (line R6/2) with a transgenic mouse that expresses a CRE regulated reporter gene. Instead of compromised CRE-dependent transcription in HD mice, we found a robust upregulation of CRE-dependent transcription in several brain regions (striatum, hippocampus, cortex). CRE-mediated transcription was also evoked by striatal forskolin infusion and by photic stimulation in HD animals. Increased cAMP response element-binding protein (CREB) phosphorylation and elevated levels of the CREB-regulated gene product, CCAAT/enhancer binding protein beta, were also found in HD mice. Significant alterations in CREB binding protein expression and localization were not observed in symptomatic R6/2 mice. Thus, rather than repressing CRE-mediated transcription, mutant huntingtin appears to facilitate transcription via a CRE-dependent mechanism in vivo. PMID- 14749424 TI - Inhibition of N-type voltage-activated calcium channels in rat dorsal root ganglion neurons by P2Y receptors is a possible mechanism of ADP-induced analgesia. AB - Patch-clamp recordings from small-diameter rat dorsal root ganglion (DRG) neurons maintained in culture demonstrated preferential inhibition by ATP of high-voltage activated, but not low-voltage-activated, Ca2+ currents (I(Ca)). The rank order of agonist potency was UTP > ADP > ATP. ATP depressed the omega-conotoxin GVIA sensitive N-type current only. Pyridoxal-5-phosphate-6-azophenyl-2',4' disulphonic acid (PPADS) and 2'-deoxy-N6-methyladenosine 3',5'-bisphosphate tetraammonium, two P2Y1 receptor antagonists, almost abolished the ATP-induced inhibition. Both patch-clamp recordings and immunocytochemistry coupled with confocal laser microscopy indicated a colocalization of functional P2X3 and P2Y1 receptors on the same DRG neurons. Because the effect of ATP was inhibited by intracellular guanosine 5'-O-(2-thiodiphosphate) or by applying a strongly depolarizing prepulse, P2Y1 receptors appear to block I(Ca) by a pathway involving the betagamma subunit of a G(q/11) protein. Less efficient buffering of the intracellular Ca2+ concentration ([Ca2+]i) by reducing the intrapipette EGTA failed to interfere with the ATP effect. Fura-2 microfluorimetry suggested that ATP raised [Ca2+]i by a Galpha-mediated release from intracellular pools and simultaneously depressed the high external potassium concentration-induced increase of [Ca2+]i by inhibiting I(Ca) via Gbetagamma. Adenosine 5'-O-(2 thiodiphosphate) inhibited dorsal root-evoked polysynaptic population EPSPs in the hemisected rat spinal cord and prolonged the nociceptive threshold on intrathecal application in the tail-flick assay. These effects were not antagonized by PPADS. Hence, P2Y receptor activation by ADP, which is generated by enzymatic degradation of ATP, may decrease the release of glutamate from DRG terminals in the spinal cord and thereby partly counterbalance the algogenic effect of ATP. PMID- 14749425 TI - Repulsive guidance molecule (RGM) gene function is required for neural tube closure but not retinal topography in the mouse visual system. AB - The establishment of topographic projections in the developing visual system depends on the spatially and temporally controlled expression of axon guidance molecules. In the developing chick tectum, the graded expression of the repulsive guidance molecule (RGM) has been proposed to be involved in controlling the topography of the retinal ganglion cell (RGC) axon termination zones along the anteroposterior axis of the tectum. We now show that there are three mouse proteins homologous to chick RGM displaying similar proteolytic processing but exhibiting differential cell-surface targeting by glycosyl phosphatidylinositol anchor addition. Two members of this gene family (mRGMa and mRGMb) are expressed in complementary patterns in the nervous system, and mRGMa is expressed prominently in the superior colliculus at the time of anteroposterior targeting of RGC axons. The third member of the family (mRGMc) is expressed almost exclusively in skeletal muscles. Functional studies in the mouse reveal a role for mRGMa in controlling cephalic neural tube closure, thus defining an unexpected role for mRGMa in early embryonic development. In contrast, mRGMa mutant mice did not exhibit defects in anteroposterior targeting of RGC axons to their stereotypic termination zones in the superior colliculus. PMID- 14749426 TI - Semaphorin3A inhibits nerve growth factor-induced sprouting of nociceptive afferents in adult rat spinal cord. AB - Increased expression of NGF after spinal cord injury induces sprouting of primary nociceptive axons. Exogenous application of NGF also results in extensive sprouting of these axons and causes chronic pain in uninjured animals. During development, semaphorin3A is thought to act as a repulsive guidance cue for NGF responsive nociceptive afferents, restricting their projections to the superficial dorsal horn. We investigated the ability of semaphorin3A to selectively reduce NGF-induced sprouting and neuropathic pain in adult rats. The chemorepulsive effect of virus-mediated semaphorin3A expression was shown to counteract the sprouting induced by NGF in a dose-dependent manner, both in vitro and in adult rat spinal cords. Coexpression of semaphorin3A and NGF at moderate to low concentrations within the adult spinal cord reduced sprouting of calcitonin gene-related peptide and substance P-containing axons compared with GFP and NGF coexpression controls. At high expression levels of NGF, there was no difference in sprouting between the semaphorin3A-treated and control groups. The distribution of endogenous primary nociceptive afferents in the spinal cord appeared to be unaffected by semaphorin3A treatment in these experiments. Behavioral assessment shows that semaphorin3A coexpression with NGF led to decreased mechanical allodynia but no significant reductions in thermal hyperalgesia. These findings demonstrate directly that mature sensory afferents maintain their responsiveness to semaphorin3A, suggesting that this molecule might be used therapeutically to control aberrant sensory sprouting involved in pain or autonomic dysfunction. PMID- 14749427 TI - Endogenous activation of mGlu5 metabotropic glutamate receptors contributes to the development of nigro-striatal damage induced by 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine in mice. AB - We combined the use of knock-out mice and subtype-selective antagonists [2-methyl 6-(phenylethynyl)pyridine (MPEP) and (E)-2-methyl-6-(2-phenylethenyl)-pyridine (SIB1893)] to examine whether endogenous activation of mGlu5 metabotropic glutamate receptors contributes to the pathophysiology of nigro-striatal damage in the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) model of parkinsonism. High doses of MPTP (four injections of 20 mg/kg, i.p., every 2 hr) induced a high mortality rate and a nearly total degeneration of the nigro-striatal pathway in wild-type mice. mGlu5 knock-out mice were less sensitive to MPTP toxicity, as shown by a higher survival and a milder nigro-striatal damage. Protection against MPTP (80 mg/kg) toxicity was also observed after MPEP injections (four injections of 5 mg/kg, i.p., 30 min before each MPTP injection). MPEP treatment did not further increase neuroprotection against 80 mg/kg of MPTP in mGlu5 knock-out mice, indicating that the drug acted by inhibiting mGlu5 receptors. In wild-type mice, MPEP was also neuroprotective when challenged against lower doses of MPTP (either 30 mg/kg, single injection, or four of 10 mg/kg injections). The action of MPEP was mimicked by SIB1893 but not by the mGlu1 receptor antagonist 7 hydroxyiminocyclopropan[b]chromen-1a-carboxylic acid ethyl ester. MPEP did not change the kinetics of 1-methyl-4-phenylpyridinium ion formation in the striatum of mice injected with MPTP. We conclude that mGlu5 receptors act as amplifiers of MPTP toxicity and that mGlu5 receptor antagonists may limit the extent of nigro striatal damage in experimental models of parkinsonism. PMID- 14749428 TI - Morphological and physiological features of a set of spinal substantia gelatinosa neurons defined by green fluorescent protein expression. AB - The spinal substantia gelatinosa (SG) is known to be involved in the manipulation of nociceptive and thermal primary afferent input; however, the interrelationships of its neuronal components are poorly understood. As a step toward expanding understanding, we took a relatively unique approach by concentrating on a set of SG neurons selectively labeled by green fluorescent protein (GFP) in a transgenic mouse. These GFP-expressing SG neurons prove to have homogenous morphological and electrophysiological properties, are systematically spaced in the SG, contain GABA, receive C-fiber primary afferent input, and upregulate c-Fos protein in response to noxious stimuli. Together, the properties established for these GFP-labeled neurons are consistent with a modular SG organization in which afferent activity related to nociception or other C-fiber signaling are subject to integration/modulation by repeating, similar circuits of neurons. PMID- 14749429 TI - Enhanced in vitro midbrain dopamine neuron differentiation, dopaminergic function, neurite outgrowth, and 1-methyl-4-phenylpyridium resistance in mouse embryonic stem cells overexpressing Bcl-XL. AB - Embryonic stem (ES) cells provide a potentially unlimited source of specialized cells for regenerative medicine. The ease of inducing stable genetic modifications in ES cells allows for in vitro manipulations to enhance differentiation into specific cell types and to optimize in vivo function of differentiated progeny in animal models of disease. We have generated mouse ES cells that constitutively express Bcl-XL, an antiapoptotic protein of Bcl-2 family. In vitro differentiation of Bcl-XL overexpressing ES (Bcl-ES) cells resulted in higher expression of genes related to midbrain dopamine (DA) neuron development and increased the number of ES-derived neurons expressing midbrain DA markers compared with differentiation of wild-type ES cells. Moreover, DA neurons derived from Bcl-ES cells were less susceptible to 1-methyl-4-phenylpyridium, a neurotoxin for DA neurons. On transplantation into parkinsonian rats, the Bcl-ES derived DA neurons exhibited more extensive fiber outgrowth and led to a more pronounced reversal of behavioral symptoms than wild-type ES-derived DA neurons. These data suggest a role for Bcl-XL during in vitro midbrain DA neuron differentiation and provide an improved system for cell transplantation in a preclinical animal model of Parkinson's disease. PMID- 14749430 TI - Translamellar disinhibition in the rat hippocampal dentate gyrus after seizure induced degeneration of vulnerable hilar neurons. AB - Longitudinally restricted axonal projections of hippocampal granule cells suggest that transverse segments of the granule cell layer may operate independently (the "lamellar" hypothesis). Longitudinal projections of excitatory hilar mossy cells could be viewed as antithetical to lamellar function, but only if longitudinal impulse flow effectively excites distant granule cells. We, therefore, determined the effect of focal granule cell discharges on granule cells located >2 mm along the longitudinal axis. During perforant pathway stimulation in urethane anesthetized rats, passive diffusion of the GABA(A) receptor antagonist bicuculline methiodide from the tip of a glass recording electrode evoked granule cell discharges and c-Fos expression in granule cells, mossy cells, and inhibitory interneurons, within a approximately 400 microm radius. This focally evoked activity powerfully suppressed distant granule cell-evoked responses recorded simultaneously approximately 2.5-4.5 mm longitudinally. Three days after kainic acid-induced status epilepticus or prolonged perforant pathway stimulation, translamellar inhibition was intact in rats with <40% hilar neuron loss but was consistently abolished after extensive (>85%) hilar cell loss. Retrograde transport of Fluoro-Gold (FG) from the rostral dentate gyrus revealed that few inhibitory interneurons were among the many retrogradely labeled hilar neurons 2.5-4.5 mm longitudinally. Although many somatostatin-positive hilar interneurons effectively transported FG from the distant septum, few of these neurons transported detectable FG from much closer hippocampal injection sites. Inhibitory basket and chandelier cells also exhibited minimal longitudinal FG transport. These findings suggest that translamellar disinhibition may result from the loss of vulnerable, longitudinally projecting mossy cells and may represent a network-level mechanism underlying postinjury hippocampal dysfunction and epileptic network hyperexcitability. PMID- 14749431 TI - Cyclin-dependent kinase 5 phosphorylates the N-terminal domain of the postsynaptic density protein PSD-95 in neurons. AB - PSD-95 (postsynaptic density 95) is a postsynaptic scaffolding protein that links NMDA receptors to the cytoskeleton and signaling molecules. The N-terminal domain of PSD-95 is involved in the synaptic targeting and clustering of PSD-95 and in the clustering of NMDA receptors at synapses. The N-terminal domain of PSD-95 contains three consensus phosphorylation sites for cyclin-dependent kinase 5 (cdk5), a proline-directed serine-threonine kinase essential for brain development and implicated in synaptic plasticity, dopamine signaling, cocaine addiction, and neurodegenerative disorders. We report that PSD-95 is phosphorylated in the N-terminal domain by cdk5 in vitro and in vivo, and that this phosphorylation is not detectable in brain lysates of cdk5-/- mice. N terminal phosphorylated PSD-95 is found in PSD fractions together with cdk5 and its activator, p35, suggesting a role for phosphorylated PSD-95 at synapses. In heterologous cells, coexpression of active cdk5 reduces the ability of PSD-95 to multimerize and to cluster neuronal ion channels, two functions attributed to the N-terminal domain of PSD-95. Consistent with these observations, the lack of cdk5 activity in cultured neurons results in larger clusters of PSD-95. In cdk5-/- cortical neurons, more prominent PSD-95 immunostained clusters are observed than in wild-type neurons. In hippocampal neurons, the expression of DNcdk5 (inactive form of cdk5) or of the triple alanine mutant (T19A, S25A, S35A) full-length PSD 95 results in increased PSD-95 cluster size. These results identify cdk5 dependent phosphorylation of the N-terminal domain of PSD-95 as a novel mechanism for regulating the clustering of PSD-95. Moreover, these observations support the possibility that cdk5-dependent phosphorylation of PSD-95 dynamically regulates the clustering of PSD-95/NMDA receptors at synapses, thus providing a possible mechanism for rapid changes in density and/or number of receptor at synapses. PMID- 14749432 TI - Action potential timing determines dendritic calcium during striatal up-states. AB - Up-states represent a key feature of synaptic integration in cortex and striatum that involves activation of many synaptic inputs. In the striatum, the sparse firing and tight control of action potential timing is in contrast to the large intracellular membrane potential depolarizations observed during the up-state. One hallmark of striatal spiny projection neurons is the delay to action potential generation in both up-states and suprathreshold depolarization by somatic current injection. By studying somatic and dendritic intracellular calcium ([Ca2+]i) transients during spontaneous up-states in cortex-striatum substantia nigra organotypic cultures, we show that the delay between up-state onset and action potential generation determines dendritic peak [Ca2+]i. Peak [Ca2+]i from single action potentials reached maximum values when action potentials were close to up-state onset and sharply decayed to near subthreshold up-state [Ca2+] levels as a function of time (tau = 47 +/- 26 msec for tertiary dendrite). Similarly, a precisely timed action potential elicited during subthreshold up-states through somatic current injection established that the delay between up-state onset and action potential generation is the critical variable that controls peak [Ca2+]i. Blocking NMDA channels internally with high intracellular Mg2+ ([Mg2+]i) (10 mm) abolished the dependency of peak [Ca2+]i on action potential timing during spontaneous up-states. Finally, high [Mg2+]i specifically blocked [Ca2+]i transients that resulted from local NMDA application in conjunction with backpropagating action potentials. We conclude that precisely timed, single action potentials during striatal up-states control peak dendritic calcium levels. We suggest that this mechanism might play an important role in synaptic plasticity of the corticostriatal pathway. PMID- 14749433 TI - Molecular basis of gap junctional communication in the CNS of the leech Hirudo medicinalis. AB - Gap junctions are intercellular channels that allow the passage of ions and small molecules between cells. In the nervous system, gap junctions mediate electrical coupling between neurons. Despite sharing a common topology and similar physiology, two unrelated gap junction protein families exist in the animal kingdom. Vertebrate gap junctions are formed by members of the connexin family, whereas invertebrate gap junctions are composed of innexin proteins. Here we report the cloning of two innexins from the leech Hirudo medicinalis. These innexins show a differential expression in the leech CNS: Hm-inx1 is expressed by every neuron in the CNS but not in glia, whereas Hm-inx2 is expressed in glia but not neurons. Heterologous expression in the paired Xenopus oocyte system demonstrated that both innexins are able to form functional homotypic gap junctions. Hm-inx1 forms channels that are not strongly gated. In contrast, Hm inx2 forms channels that are highly voltage-dependent; these channels demonstrate properties resembling those of a double rectifier. In addition, Hm-inx1 and Hm inx2 are able to cooperate to form heterotypic gap junctions in Xenopus oocytes. The behavior of these channels is primarily that predicted from the properties of the constituent hemichannels but also demonstrates evidence of an interaction between the two. This work represents the first demonstration of a functional gap junction protein from a Lophotrochozoan animal and supports the hypothesis that connexin-based communication is restricted to the deuterostome clade. PMID- 14749434 TI - The activation mechanism of alpha1 homomeric glycine receptors. AB - The glycine receptor mediates fast synaptic inhibition in the spinal cord and brainstem. Its activation mechanism is not known, despite the physiological importance of this receptor and the fact that it can serve as a prototype for other homopentameric channels. We analyzed single-channel recordings from rat recombinant alpha1 glycine receptors by fitting different mechanisms simultaneously to sets of sequences of openings at four glycine concentrations (10-1000 microm). The adequacy of the mechanism and the rate constants thus fitted was judged by examining how well these described the observed dwell-time distributions, open-shut correlation, and single-channel P(open) dose-response curve. We found that gating efficacy increased as more glycine molecules bind to the channel, but maximum efficacy was reached when only three (of five) potential binding sites are occupied. Successive binding steps are not identical, implying that binding sites can interact while the channel is shut. These interactions can be interpreted in the light of the topology of the binding sites within a homopentamer. PMID- 14749435 TI - Production of 5alpha-reduced neurosteroids is developmentally regulated and shapes GABA(A) miniature IPSCs in lamina II of the spinal cord. AB - In lamina II of the spinal dorsal horn, synaptic inhibition mediated by ionotropic GABA(A) and glycine receptors contributes to the integration of peripheral nociceptive messages. Whole-cell patch-clamp recordings were performed from lamina II neurons in spinal cord slices to study the properties of miniature IPSCs (mIPSCs) mediated by activation of GABA(A) and glycine receptors in immature (<30 d) and adult rats. Blockade of neurosteroidogenesis by 1-(2 chlorophenyl)-N-methyl-N-(1-methylpropyl)-3-isoquinoline carboxamide (PK11195), an inhibitor of the peripheral benzodiazepine receptor (PBR), or finasteride, which blocks 5alpha-reductase, accelerated the decay kinetics of GABA(A) receptor mediated mIPSCs in immature, but not in adult animals. Glycine receptor-mediated mIPSCs remained unaffected under these conditions. These results suggest the presence of a tonic production of 5alpha-reduced neurosteroids in young rats that confers slow decay kinetics to GABA(A) mIPSCs. At all of the ages, selective stimulation of PBR by diazepam in the presence of flumazenil prolonged GABA(A) mIPSCs in a PK11195- and finasteride-sensitive manner. This condition also increased the proportion of mixed GABA(A)/glycine mIPSCs in the immature animals and led to the reappearance of mixed GABA(A)/glycine mIPSCs in the adult. Our results might point to an original mechanism by which the strength of synaptic inhibition can be adjusted locally in the CNS during development and under physiological and/or pathological conditions by controlling the synthesis of endogenous 5alpha-reduced neurosteroids. PMID- 14749436 TI - Postsynaptic density 95 controls AMPA receptor incorporation during long-term potentiation and experience-driven synaptic plasticity. AB - The regulated delivery of AMPA-type glutamate receptors (AMPARs) to synapses is an important mechanism underlying synaptic plasticity. Here, we ask whether the synaptic scaffolding protein PSD-95 (postsynaptic density 95) participates in AMPAR incorporation during two forms of synaptic plasticity. In hippocampal slice cultures, the expression of PSD-95-green fluorescent protein (PSD-95-GFP) increases AMPAR currents by selectively delivering glutamate receptor 1 (GluR1) containing receptors to synapses, thus mimicking long-term potentiation (LTP). Mutational analysis shows that the N terminal of PSD-95 including the first two PDZ [PSD-95/Discs large (Dlg)/zona occludens-1 (ZO-1)] domains is necessary and sufficient to mediate this effect. Further supporting a role in synaptic plasticity, wild-type PSD-95 occludes LTP and dominant negative forms block LTP. Moreover, we demonstrate that PSD-95 also participates in AMPAR delivery during experience-driven plasticity in vivo. In the barrel cortex from experience deprived animals, the expression of PSD-95-GFP selectively increases AMPAR currents, mimicking experience-driven plasticity. In nondeprived animals, PSD-95 GFP produces no additional potentiation, indicating common mechanisms between PSD 95-mediated potentiation and experience-driven synaptic strengthening. A dominant negative form of PSD-95 blocks experience-driven potentiation of synapses. Pharmacological analysis in slice cultures reveals that PSD-95 acts downstream of other signaling pathways involved in LTP. We conclude that PSD-95 controls activity-dependent AMPAR incorporation at synapses via PDZ interactions not only during LTP in vitro but also during experience-driven synaptic strengthening by natural stimuli in vivo. PMID- 14749437 TI - Phox2a gene, A6 neurons, and noradrenaline are essential for development of normal respiratory rhythm in mice. AB - Although respiration is vital to the survival of all mammals from the moment of birth, little is known about the genetic factors controlling the prenatal maturation of this physiological process. Here we investigated the role of the Phox2a gene that encodes for a homeodomain protein involved in the generation of noradrenergic A6 neurons in the maturation of the respiratory network. First, comparisons of the respiratory activity of fetuses delivered surgically from heterozygous Phox2a pregnant mice on gestational day 18 showed that the mutants had impaired in vivo ventilation, in vitro respiratory-like activity, and in vitro respiratory responses to central hypoxia and noradrenaline. Second, pharmacological studies on wild-type neonates showed that endogenous noradrenaline released from pontine A6 neurons potentiates rhythmic respiratory activity via alpha1 medullary adrenoceptors. Third, transynaptic tracing experiments in which rabies virus was injected into the diaphragm confirmed that A6 neurons were connected to the neonatal respiratory network. Fourth, blocking the alpha1 adrenoceptors in wild-type dams during late gestation with daily injections of the alpha1 adrenoceptor antagonist prazosin induced in vivo and in vitro neonatal respiratory deficits similar to those observed in Phox2a mutants. These results suggest that noradrenaline, A6 neurons, and the Phox2a gene, which is crucial for the generation of A6 neurons, are essential for development of normal respiratory rhythm in neonatal mice. Metabolic noradrenaline disorders occurring during gestation therefore may induce neonatal respiratory deficits, in agreement with the catecholamine anomalies reported in victims of sudden infant death syndrome. PMID- 14749438 TI - Polymorphisms in the taste receptor gene (Tas1r3) region are associated with saccharin preference in 30 mouse strains. AB - The results of recent studies suggest that the mouse Sac (saccharin preference) locus is identical to the Tas1r3 (taste receptor) gene. The goal of this study was to identify Tas1r3 sequence variants associated with saccharin preference in a large number of inbred mouse strains. Initially, we sequenced approximately 6.7 kb of the Tas1r3 gene and its flanking regions from six inbred mouse strains with high and low saccharin preference, including the strains in which the Sac alleles were described originally (C57BL/6J, Sac(b); DBA/2J, Sac(d)). Of the 89 sequence variants detected among these six strains, eight polymorphic sites were significantly associated with preferences for 1.6 mm saccharin. Next, each of these eight variant sites were genotyped in 24 additional mouse strains. Analysis of the genotype-phenotype associations in all 30 strains showed the strongest association with saccharin preference at three sites: nucleotide (nt) -791 (3 bp insertion/deletion), nt +135 (Ser45Ser), and nt +179 (Ile60Thr). We measured Tas1r3 gene expression, transcript size, and T1R3 immunoreactivity in the taste tissue of two inbred mouse strains with different Tas1r3 haplotypes and saccharin preferences. The results of these experiments suggest that the polymorphisms associated with saccharin preference do not act by blocking gene expression, changing alternative splicing, or interfering with protein translation in taste tissue. The amino acid substitution (Ile60Thr) may influence the ability of the protein to form dimers or bind sweeteners. Here, we present data for future studies directed to experimentally confirm the function of these polymorphisms and highlight some of the difficulties of identifying specific DNA sequence variants that underlie quantitative trait loci. PMID- 14749439 TI - Estrogen-induced mu-opioid receptor internalization in the medial preoptic nucleus is mediated via neuropeptide Y-Y1 receptor activation in the arcuate nucleus of female rats. AB - The endogenous peptides beta-endorphin (beta-END) and neuropeptide Y (NPY) have been implicated in regulating sexual receptivity. Both beta-END and NPY systems are activated by estrogen and inhibit female sexual receptivity. The initial estrogen-induced sexual nonreceptivity is correlated with the activation and internalization of mu-opioid receptors (MORs), in the medial preoptic nucleus (MPN). Progesterone reverses the estrogen-induced activation/internalization of MOR and induces the sexual receptive behavior lordosis. To determine whether NPY and endogenous opioids interact, we tested the hypothesis that estrogen-induced MOR activation is mediated through NPY-Y1 receptor (Y1R) activation. Retrograde tract tracing demonstrated Y1Ron beta-END neurons that projected to the MPN. Sex steroid modulation of MOR in the MPN acts through NPY and the Y1R. Estradiol administration or intracerebroventricular injection of NPY activated/internalized Y1R in the arcuate nucleus and MOR in the MPN of ovariectomized (OVX) rats. Moreover, the selective Y1R agonist [Leu31, Pro34]-Neuropeptide Y (LPNY) internalized MOR in the MPN of OVX rats. The Y1R antagonist (Cys31, Nva34) Neuropeptide Y (27-36)2 prevented estrogen-induced Y1R and MOR activation/internalization. NPY reversed the progesterone blockade of estradiol induced Y1R and MOR internalization in the arcuate nucleus and MPN, respectively. Behaviorally, LPNY inhibited estrogen plus progesterone-induced lordosis, and the MOR-selective antagonist D-Phe-Cys-Tyr-d-Trp-Orn-Thr-Pen-Thr amide reversed LPNY induced inhibition of lordosis. These results suggest that a sequential sex steroid activation of NPY and MOR circuits regulates sexual receptivity. PMID- 14749440 TI - Shrinkage of the entorhinal cortex over five years predicts memory performance in healthy adults. AB - Lesions in the hippocampus (HC), the entorhinal cortex (EC), and the prefrontal cortex (PFC) are associated with impairment of episodic memory; reduced HC volume is linked to memory declines in dementia; and decline in EC volume predicts progression from mild cognitive impairment to dementia. However, in healthy adults, the relationship between memory and regional volumes is unclear, and no data are available on the relationship of longitudinal regional shrinkage to memory performance in a cognitively intact population. The objective of this study was to examine whether shrinkage of the EC, HC, and PFC over a 5 year period can predict declarative memory performance in healthy adults. The volumes of three brain regions were measured on magnetic resonance images that were acquired twice, 5 years apart. Multiple measures of episodic memory were administered at follow-up. Results indicated that the volume of HC and PFC (but not EC) correlated with age at baseline and follow-up. However, after age differences in memory were taken into account, none of the regional volumes was associated with memory performance at follow-up. In contrast, greater annual rate of shrinkage in EC (but not HC or PFC) predicted poorer memory performance. Thus, in a healthy and educated population, even mild age-related shrinkage of the EC may be a sensitive predictor of memory decline. PMID- 14749441 TI - Perceptual correlates of nociceptive long-term potentiation and long-term depression in humans. AB - Long-term potentiation (LTP) and long-term depression (LTD) of synaptic strength are ubiquitous mechanisms of synaptic plasticity, but their functional relevance in humans remains obscure. Here we report that a long-term increase in perceived pain to electrical test stimuli was induced by high-frequency electrical stimulation (HFS) (5 x 1 sec at 100 Hz) of peptidergic cutaneous afferents (27% above baseline, undiminished for >3 hr). In contrast, a long-term decrease in perceived pain (27% below baseline, undiminished for 1 hr) was induced by low frequency stimulation (LFS) (17 min at 1 Hz). Pain testing with punctate mechanical probes (200 microm diameter) in skin adjacent to the HFS-LFS conditioning skin site revealed a marked twofold to threefold increase in pain sensitivity (secondary hyperalgesia, undiminished for >3 hr) after HFS but also a moderate secondary hyperalgesia (30% above baseline) after strong LFS. Additionally, HFS but not LFS caused pain to light tactile stimuli in adjacent skin (allodynia). In summary, HFS and LFS stimulus protocols that induce LTP or LTD in spinal nociceptive pathways in animal experiments led to similar LTP- and LTD-like changes in human pain perception (long-term hyperalgesia or hypoalgesia) mediated by the conditioned pathway. Additionally, secondary hyperalgesia and allodynia in adjacent skin induced by the HFS protocol and, to a minor extent, also by the LFS protocol, suggested that these perceptual changes encompassed an LTP-like heterosynaptic facilitation of adjacent nociceptive pathways by a hitherto unknown mechanism. PMID- 14749442 TI - Dopamine neurons mediate a fast excitatory signal via their glutamatergic synapses. AB - Dopamine neurons are thought to convey a fast, incentive salience signal, faster than can be mediated by dopamine. A resolution of this paradox may be that midbrain dopamine neurons exert fast excitatory actions. Using transgenic mice with fluorescent dopamine neurons, in which the axonal projections of the neurons are visible, we made horizontal brain slices encompassing the mesoaccumbens dopamine projection. Focal extracellular stimulation of dopamine neurons in the ventral tegmental area evoked dopamine release and early monosynaptic and late polysynaptic excitatory responses in postsynaptic nucleus accumbens neurons. Local superfusion of the ventral tegmental area with glutamate, which should activate dopamine neurons selectively, produced an increase in excitatory synaptic events. Local superfusion of the ventral tegmental area with the D2 agonist quinpirole, which should increase the threshold for dopamine neuron activation, inhibited the early response. So dopamine neurons make glutamatergic synaptic connections to accumbens neurons. We propose that dopamine neuron glutamatergic transmission may be the initial component of the incentive salience signal. PMID- 14749444 TI - Presynaptic localization of neprilysin contributes to efficient clearance of amyloid-beta peptide in mouse brain. AB - A local increase in amyloid-beta peptide (Abeta) is closely associated with synaptic dysfunction in the brain in Alzheimer's disease. Here, we report on the catabolic mechanism of Abeta at the presynaptic sites. Neprilysin, an Abeta degrading enzyme, expressed by recombinant adeno-associated viral vector-mediated gene transfer, was axonally transported to presynaptic sites through afferent projections of neuronal circuits. This gene transfer abolished the increase in Abeta levels in the hippocampal formations of neprilysin-deficient mice and also reduced the increase in young mutant amyloid precursor protein transgenic mice. In the latter case, Abeta levels in the hippocampal formation contralateral to the vector-injected side were also significantly reduced as a result of transport of neprilysin from the ipsilateral side, and in both sides soluble Abeta was degraded more efficiently than insoluble Abeta. Furthermore, amyloid deposition in aged mutant amyloid precursor protein transgenic mice was remarkably decelerated. Thus, presynaptic neprilysin has been demonstrated to degrade Abeta efficiently and to retard development of amyloid pathology. PMID- 14749445 TI - Optical recording of action potentials with second-harmonic generation microscopy. AB - Nonlinear microscopy has proven to be essential for neuroscience investigations of thick tissue preparations. However, the optical recording of fast (approximately 1 msec) cellular electrical activity has never until now been successfully combined with this imaging modality. Through the use of second harmonic generation microscopy of primary Aplysia neurons in culture labeled with 4-[4-(dihexylamino)phenyl][ethynyl]-1-(4-sulfobutyl)pyridinium (inner salt), we optically recorded action potentials with 0.833 msec temporal and 0.6 microm spatial resolution on soma and neurite membranes. Second-harmonic generation response as a function of change in membrane potential was found to be linear with a signal change of approximately 6%/100 mV. The signal-to-noise ratio was approximately 1 for single-trace action potential recordings but was readily increased to approximately 6-7 with temporal averaging of approximately 50 scans. Photodamage was determined to be negligible by observing action potential characteristics, cellular resting potential, and gross cellular morphology during and after laser illumination. High-resolution (micrometer scale) optical recording of membrane potential activity by previous techniques has been limited to imaging depths an order of magnitude less than nonlinear methods. Because second-harmonic generation is capable of imaging up to approximately 400 microm deep into intact tissue with submicron resolution and little out-of-focus photodamage or bleaching, its ability to record fast electrical activity should prove valuable to future electrophysiology studies. PMID- 14749446 TI - How industry is approaching the search for new diagnostic markers and biomarkers. AB - In the diagnostic and the pharmaceutical industry there is a constant need for new diagnostic markers and biomarkers with improved sensitivity and specificity. During the last 5 years, only a few novel diagnostic markers have been introduced into the market. Proteomics technologies are now offering unique chances to identify new candidate markers. Before a marker can be introduced into the market, three successive developmental phases have to be completed: the discovery phase, in which a variety of proteomics technologies are applied to identify marker candidates; the prototype developmental phase, in which immunological assays are established and validated in defined sample collectives; and finally the product development phase, with assay formats suitable for automated platforms. The hurdles that a potential candidate marker has to pass in each developmental phase before reaching the market are considerable. The costs are increasing from phase to phase, and in industry a number of questions concerning the medical need and the potential return on investment have to be answered before a proteomics discovery project is started. In this review, we will cover aspects of all three developmental phases including the repertoire of discovery tools for protein separation as well as giving an outline of modern principles of mass spectrometry for the identification of proteins. PMID- 14749447 TI - Human metapneumovirus--an important new respiratory virus. PMID- 14749443 TI - Morphological effects of estrogen on cholinergic neurons in vitro involves activation of extracellular signal-regulated kinases. AB - In the present study, we examined the ability of estrogen to enhance cholinergic neurite arborization in vitro and identified the signal transduction cascade associated with this effect. Basal forebrain primordia collected from rat pups on postnatal day 1 were cultured for 2 weeks and then treated with 5 nm 17beta estradiol for 24 hr. Cholinergic neurons were identified immunocytochemically with an antibody against the vesicular acetylcholine transporter and digitally photographed. Morphological analysis indicated that female cultures respond to estrogen treatment with an increase in total neurite length per neuron (4.5-fold over untreated controls) and in total branch segment number per neuron (2.3-fold over controls). In contrast, there was no change in total neurite length per neuron in male cultures, and we also observed a decrease in total branch segment number per neuron (0.5-fold below controls). Detailed histograms indicated that estrogen increases primary and secondary branch length and number and also increases terminal neuritic branches to the seventh order in female cultures. In a second set of experiments, we investigated the signal transduction cascade involved in this response, and found that an upstream extracellular signal regulated kinase (ERK) inhibitor blocked the ability of estrogen to enhance outgrowth in female cultures. Our study provides strong evidence in support of the fact that the ERK pathway is required for estrogen-induced structural plasticity in the cholinergic system of female rats. Understanding the intracellular processes that underlie the response of cholinergic neurons to estrogen provides a necessary step in elucidating how cholinergic neurons can be particularly susceptible to degeneration in postmenopausal women. PMID- 14749448 TI - Diagnosing ventilator-associated pneumonia. PMID- 14749449 TI - A clearer view of effusive-constrictive pericarditis. PMID- 14749450 TI - Preventing foodborne disease--what clinicians can do. PMID- 14749451 TI - Hypogonadism in elderly men--what to do until the evidence comes. PMID- 14749452 TI - Human metapneumovirus and lower respiratory tract disease in otherwise healthy infants and children. AB - BACKGROUND: We sought to determine the role of human metapneumovirus in lower respiratory tract illness in previously healthy infants and children. METHODS: We tested nasal-wash specimens, obtained over a 25-year period from otherwise healthy children presenting with acute respiratory tract illness, for human metapneumovirus. RESULTS: A viral cause other than human metapneumovirus was determined for 279 of 687 visits for acute lower respiratory tract illness (41 percent) by 463 children in a population of 2009 infants and children prospectively seen from 1976 to 2001. There were 408 visits for lower respiratory tract illness by 321 children for which no cause was identified. Of these 321 children, specimens from 248 were available. Forty-nine of these 248 specimens (20 percent) contained human metapneumovirus RNA or viable virus. Thus, 20 percent of all previously virus-negative lower respiratory tract illnesses were attributable to human metapneumovirus, which means that 12 percent of all lower respiratory tract illnesses in this cohort were most likely due to this virus. The mean age of human metapneumovirus-infected children was 11.6 months, the male:female ratio was 1.8:1, 78 percent of illnesses occurred between December and April, and the hospitalization rate was 2 percent. The virus was associated with bronchiolitis in 59 percent of cases, pneumonia in 8 percent, croup in 18 percent, and an exacerbation of asthma in 14 percent. We also detected human metapneumovirus in 15 percent of samples from 261 patients with upper respiratory tract infection but in only 1 of 86 samples from asymptomatic children. CONCLUSIONS: Human metapneumovirus infection is a leading cause of respiratory tract infection in the first years of life, with a spectrum of disease similar to that of respiratory syncytial virus. PMID- 14749453 TI - Soluble triggering receptor expressed on myeloid cells and the diagnosis of pneumonia. AB - BACKGROUND: The diagnosis and treatment of bacterial pneumonia in patients who are receiving mechanical ventilation remain a difficult challenge. The triggering receptor expressed on myeloid cells (TREM-1) is a member of the immunoglobulin superfamily, and its expression on phagocytes is specifically up-regulated by microbial products. The presence of soluble TREM-1 (sTREM-1) in bronchoalveolar lavage fluid from patients receiving mechanical ventilation may be an indicator of pneumonia. METHODS: We conducted a prospective study of 148 patients receiving mechanical ventilation in whom infectious pneumonia was suspected. A rapid immunoblot technique was used to measure sTREM-1 in bronchoalveolar-lavage fluid. Two independent intensivists who were unaware of the results of the sTREM-1 assay determined whether community-acquired pneumonia and ventilator-associated pneumonia were present or absent. RESULTS: The final diagnosis was community acquired pneumonia in 38 patients, ventilator-associated pneumonia in 46 patients, and no pneumonia in 64 patients. The presence of sTREM-1 by itself was more accurate than any clinical findings or laboratory values in identifying the presence of bacterial or fungal pneumonia (likelihood ratio, 10.38; sensitivity, 98 percent; specificity, 90 percent). In multiple logistic-regression analysis, the presence of sTREM-1 was the strongest independent predictor of pneumonia (odds ratio, 41.5). CONCLUSIONS: In patients receiving mechanical ventilation, rapid detection of sTREM-1 in bronchoalveolar-lavage fluid may be useful in establishing or excluding the diagnosis of bacterial or fungal pneumonia. PMID- 14749455 TI - Effusive-constrictive pericarditis. AB - BACKGROUND: Effusive-constrictive pericarditis is an uncommon pericardial syndrome characterized by concomitant tamponade, caused by tense pericardial effusion, and constriction, caused by the visceral pericardium. We conducted a prospective study of its clinical evolution and management. METHODS: From 1986 through 2001, all patients with effusive-constrictive pericarditis were prospectively evaluated. Combined pericardiocentesis and cardiac catheterization were performed in all patients, and pericardiectomy was performed in those with persistent constriction. Follow-up ranged from 1 month to 15 years (median, 7 years). RESULTS: A total of 1184 patients with pericarditis were evaluated, 218 of whom had tamponade. Of these 218, 190 underwent combined pericardiocentesis and catheterization. Fifteen of these patients had effusive-constrictive pericarditis and were included in the study. All patients presented with clinical tamponade; however, concomitant constriction was recognized in only seven patients. At catheterization, all patients had elevated intrapericardial pressure (median, 12 mm Hg; interquartile range, 7 to 18) and elevated right atrial and end-diastolic right and left ventricular pressures. After pericardiocentesis, the intrapericardial pressure decreased (median value, -5 mm Hg; interquartile range, -5 to 0), whereas right atrial and end-diastolic right and left ventricular pressures, although slightly reduced, remained elevated, with a dip-plateau morphology. The causes were diverse, and death was mainly related to the underlying disease. Pericardiectomy was required in seven patients, all of whom had involvement of the visceral pericardium. Three patients had spontaneous resolution. CONCLUSIONS: Effusive-constrictive pericarditis is an uncommon pericardial syndrome that may be missed in some patients who present with tamponade. Although evolution to persistent constriction is frequent, idiopathic cases may resolve spontaneously. In our opinion, extensive epicardiectomy is the procedure of choice in patients requiring surgery. PMID- 14749454 TI - The effects of strontium ranelate on the risk of vertebral fracture in women with postmenopausal osteoporosis. AB - BACKGROUND: Osteoporotic structural damage and bone fragility result from reduced bone formation and increased bone resorption. In a phase 2 clinical trial, strontium ranelate, an orally active drug that dissociates bone remodeling by increasing bone formation and decreasing bone resorption, has been shown to reduce the risk of vertebral fractures and to increase bone mineral density. METHODS: To evaluate the efficacy of strontium ranelate in preventing vertebral fractures in a phase 3 trial, we randomly assigned 1649 postmenopausal women with osteoporosis (low bone mineral density) and at least one vertebral fracture to receive 2 g of oral strontium ranelate per day or placebo for three years. We gave calcium and vitamin D supplements to both groups before and during the study. Vertebral radiographs were obtained annually, and measurements of bone mineral density were performed every six months. RESULTS: New vertebral fractures occurred in fewer patients in the strontium ranelate group than in the placebo group, with a risk reduction of 49 percent in the first year of treatment and 41 percent during the three-year study period (relative risk, 0.59; 95 percent confidence interval, 0.48 to 0.73). Strontium ranelate increased bone mineral density at month 36 by 14.4 percent at the lumbar spine and 8.3 percent at the femoral neck (P<0.001 for both comparisons). There were no significant differences between the groups in the incidence of serious adverse events. CONCLUSIONS: Treatment of postmenopausal osteoporosis with strontium ranelate leads to early and sustained reductions in the risk of vertebral fractures. PMID- 14749456 TI - Prevention of hepatitis A with the hepatitis A vaccine. PMID- 14749458 TI - Images in clinical medicine. Retroperitoneal sarcoma. PMID- 14749457 TI - Risks of testosterone-replacement therapy and recommendations for monitoring. PMID- 14749459 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 4-2004. A nine-month-old boy with an orbital rhabdomyosarcoma. PMID- 14749460 TI - Strontium ranelate--a novel therapy for osteoporosis or a permutation of the same? PMID- 14749461 TI - The mental health maze and the call for transformation. PMID- 14749462 TI - NIH report card. PMID- 14749463 TI - Prognostic value of myeloperoxidase in patients with chest pain. PMID- 14749464 TI - Hospital use and survival among Veterans Affairs beneficiaries. PMID- 14749465 TI - Gout. PMID- 14749466 TI - Case 5-2003: a 16-year-old girl with a rash and chest pain. PMID- 14749467 TI - Bath toys--a source of gastrointestinal infection. PMID- 14749468 TI - Images in clinical medicine. Hypoglossal-nerve palsy. PMID- 14749469 TI - Selective DNA amplification from complex genomes using universal double-sided adapters. AB - There is a rapidly developing need for new technologies to amplify millions of different targets from genomic DNA for high throughput genotyping and population gene-sequencing from diverse species. Here we describe a novel approach for the specific selection and amplification of genomic DNA fragments of interest that eliminates the need for costly and time consuming synthesis and testing of potentially millions of amplicon-specific primers. This technique relies upon Type IIs restriction enzyme digestion of genomic DNA and ligation of the fragments to double-sided adapters to form closed-circular DNA molecules. The novel use of double-sided adapters, assembled through the combinatorial use of two small universal sets of oligonucleotide building blocks, provides greater selection capacity by utilizing both sides of the adapter in a sequence-specific ligation event. As demonstrated, formation of circular structures results in protection of the desired molecules from nuclease treatment and enables a level of selectivity high enough to isolate single, or multiple, pre-defined fragments from the human genome when digested at over five million sites. Priming sites incorporated into the adapter allows the utilization of a common pair of primers for the amplification of any adapter-captured DNA fragment of interest. PMID- 14749470 TI - The discovery of a new structural class of cyclin-dependent kinase inhibitors, aminoimidazo[1,2-a]pyridines. AB - The protein kinase family represents an enormous opportunity for drug development. However, the current limitation in structural diversity of kinase inhibitors has complicated efforts to identify effective treatments of diseases that involve protein kinase signaling pathways. We have identified a new structural class of protein serine/threonine kinase inhibitors comprising an aminoimidazo[1,2-a]pyridine nucleus. In this report, we describe the first successful use of this class of aza-heterocycles to generate potent inhibitors of cyclin-dependent kinases that compete with ATP for binding to a catalytic subunit of the protein. Co-crystal structures of CDK2 in complex with lead compounds reveal a unique mode of binding. Using this knowledge, a structure-based design approach directed this chemical scaffold toward generating potent and selective CDK2 inhibitors, which selectively inhibited the CDK2-dependent phosphorylation of Rb and induced caspase-3-dependent apoptosis in HCT 116 tumor cells. The discovery of this new class of ATP-site-directed protein kinase inhibitors, aminoimidazo[1,2-a]pyridines, provides the basis for a new medicinal chemistry tool to be used in the search for effective treatments of cancer and other diseases that involve protein kinase signaling pathways. PMID- 14749471 TI - Signal transducer and activator of transcription 3 (STAT3) activation in prostate cancer: Direct STAT3 inhibition induces apoptosis in prostate cancer lines. AB - Signal transducers and activators of transcription (STAT) were originally discovered as components of cytokine signal transduction pathways. Persistent activation of one STAT, STAT3, is a common feature of prostate cancer. Activated STAT3 was found in pathology specimens obtained from prostatectomy in the cancerous areas but not in the normal margins. Because the activation of STAT3 is mediated by the action of an upstream Janus kinase (JAK) kinase, usually JAK1 or JAK2, the activation step for STAT3 might itself be a target for therapy in prostate cancer. However, the redundancy of upstream kinases may make this strategy unreliable for therapy. To develop molecular targets for prostate cancer treatment, JAK kinase and STAT3 inhibition of two prostate cancer lines were compared. DU145 and NRP-154 cells were treated with JAK kinase inhibitors, analyzed for onset of apoptosis, and measured by annexin V binding and propidium iodide uptake. Activation of caspases in the cells was determined by measuring cleaved caspase-3 following treatment. For determining the effect on mitochondrial membrane depolarization that accompanies apoptosis, the fluorescent dye JC-1 was used. STAT3 was specifically inhibited by transfecting either a dominant-negative (DN) STAT3 plasmid or antisense STAT3 oligonucleotides into the cells. To look for reduction in STAT3 levels within cells, fixed and permeabilized prostate cancer cells were stained with antibody to STAT3. We found that more than one JAK kinase is involved in STAT3 activation in prostate cancer lines. AG490 (JAK2 specific) induced apoptosis in DU145 but not in NRP-154 prostate cancer lines, whereas piceatannol (JAK1 specific) induced apoptosis in NRP-154 but not in DU145 cells. Next, we demonstrated efficacy of specific STAT3 inhibitors in prostate cancer lines. Both induction of apoptosis and reduction in intracellular STAT3 protein were observed following treatment with antisense STAT3 oligonucleotides, while transfection of a DN-STAT3 plasmid into both prostate cancer cell lines resulted in loss of viability and onset of apoptosis. We conclude that STAT3-specific inhibitors, rather than JAK kinase-specific inhibitors, should be more useful therapeutically in treating androgen-resistant prostate cancer and that STAT3 is an appropriate target in the treatment of prostate cancer. PMID- 14749472 TI - Phosphorylation of extracellular signal-regulated kinase 1 and 2, protein kinase B, and signal transducer and activator of transcription 3 are differently inhibited by an epidermal growth factor receptor inhibitor, EKB-569, in tumor cells and normal human keratinocytes. AB - EKB-569 is an irreversible inhibitor of epidermal growth factor receptor (EGF-R) tyrosine kinase. It inhibits EGF-induced phosphorylation of EGF-R and the growth of tumors that overexpress EGF-R in animal models. In clinical trials, EKB-569 and all other EGF-R inhibitors cause skin rashes. To understand the latter phenomenon, the effect of EKB-569 on EGF-R as well as downstream signaling to phosphoinositide 3-kinase-protein kinase B (AKT), extracellular signal-regulated kinase 1 and 2 (ERK1/2), or signal transducer and activator of transcription 3 (STAT3) pathways were compared in tumor cell lines and normal human keratinocytes (NHEK) grown in tissue culture. Tumor cell lines that have high (A431 epidermoid and MDA-468 breast carcinomas) and low (MCF-7 breast carcinoma) expression of EGF R were used. NHEK cells express at least 15-fold less EGF-R than A431 cells. EKB 569 was a potent inhibitor of proliferation in NHEK, A431, and MDA-468 cells (IC(50) = 61, 125, and 260 nM, respectively) but not MCF-7 cells (IC(50) = 3600 nM). EKB-569 was also a potent inhibitor of EGF-induced phosphorylated EGF-R (pEGF-R) in A431 and NHEK cells (IC(50) = 20-80 nM). The reduction in pEGF-R paralleled inhibition of phosphotyrosine-705 STAT3, while the inhibition of phosphorylated AKT and phosphorylated ERK1/2 occurred at higher concentrations of EKB-569 (75-500 nM) in both A431 and NHEK cells. The effects were specific because EKB-569 did not inhibit the nuclear factor-kappaB pathway. It is proposed that skin toxicity associated with EKB-569 is due to inhibition of EGF-R signaling. Downstream signal transduction markers, particularly the activation status of STAT3, may be useful surrogate markers to guide clinical development of EGF-R inhibitors. PMID- 14749473 TI - CXCR4 antagonist inhibits stromal cell-derived factor 1-induced migration and invasion of human pancreatic cancer. AB - The stromal cell-derived factor-1 (SDF-1)/CXCR4 system is implicated in various instances of cell migration in mammals, including the migration of lymphocytes and the formation of metastases. We have recently synthesized a potent novel CXCR4 antagonist, TN14003. The purpose of this study was to investigate the role of SDF-1/CXCR4 axis in the pancreatic cancer metastasis via cell migration and invasion, and the inhibitory effect of TN14003 on pancreatic cancer cell metastasis. The expression of CXCR4 was detected in six pancreatic cancer cell lines by Western blotting and immunocytochemistry. In migration and invasion assays, SDF-1 stimulated both migration and invasion of cancer cells in a dose dependent manner. The maximal effect of SDF-1 was observed at 100 ng/ml. SDF-1 induced migration and invasion of cancer cells were completely blocked by 100 nM TN14003. The stimulatory effect of SDF-1 on cancer migration and the inhibitory effect of TN14003 were mediated via the alteration in phosphorylation of mitogen activated protein kinases. Treatment of cancer cells with 100 ng/ml SDF-1 resulted in a significant increase of actin polymerization, which was reduced by 100 nM TN14003. SDF-1 enhanced cancer cell adhesion to laminin, which was not reversed by TN14003. Taken together, SDF-1/CXCR4 axis is involved in pancreatic cancer metastasis through migration and invasion. The small molecule antagonists against CXCR4 such as TN14003 might be an effective anti-metastatic agent for pancreatic cancer. PMID- 14749474 TI - Induction of redox imbalance and apoptosis in multiple myeloma cells by the novel triterpenoid 2-cyano-3,12-dioxoolean-1,9-dien-28-oic acid. AB - The synthetic oleanane triterpenoid 2-cyano-3,12-dioxoolean-1,9-dien-28-oic acid (CDDO) and its chemical derivatives induce differentiation and apoptosis of human leukemia cells. The precise mechanisms responsible for the effects of CDDO, however, remain unclear. In the present study, we examined the effects of CDDO and its C-28 imidazolide ester (CDDO-Im) on apoptosis of multiple myeloma (MM) cells. The results show that both CDDO and CDDO-Im are potent inducers of MM cell apoptosis and that CDDO-Im is more active than CDDO. CDDO-Im treatment was associated with (a) depletion of glutathione, (b) increases in reactive oxygen species, (c) a reduction of the Fas-associated death domain (FADD)-like interleukin-1-converting enzyme (FLICE) inhibitory protein, (d) activation of caspase-8, and (e) a decrease of the mitochondrial transmembrane potential. The reducing agents, N-acetyl-L-cysteine, DTT, and catalase inhibited each of these CDDO-Im-induced proapoptotic signals. Inhibition of caspase-8 with z-IETD-fmk also abrogated CDDO-Im-induced decreases of the mitochondrial transmembrane potential and inhibited apoptosis. These results demonstrate that CDDO-Im disrupts intracellular redox balance and thereby activates the extrinsic caspase 8-dependent apoptotic pathway. We further show that CDDO-Im induces apoptosis of primary MM cells at submicromolar concentrations and that MM cells are more sensitive to this agent than normal bone marrow mononuclear cells. These results suggest that CDDO compounds have potential as new agents for the treatment of MM. PMID- 14749475 TI - Biological characterization of MLN944: a potent DNA binding agent. AB - MLN944 (XR5944) is a novel bis-phenazine that has demonstrated exceptional efficacy against a number of murine and human tumor models. The drug was reported originally as a dual topoisomerase I/II poison, but a precise mechanism of action for this compound remains to be determined. Several lines of evidence, including the marginal ability of MLN944 to stabilize topoisomerase-dependent cleavage, and the sustained potency of MLN944 in mammalian cells with reduced levels of both topoisomerases, suggest that other activities of the drug exist. In this study, we show that MLN944 intercalates into DNA, but has no effect on the catalytic activity of either topoisomerase I or II. MLN944 displays no significant ability to stimulate DNA scission mediated by either topoisomerase I or II compared with camptothecin or etoposide, respectively. In addition, yeast genetic models also point toward a topoisomerase-independent mechanism of action. To examine cell cycle effects, synchronized human HCT116 cells were treated with MLN944, doxorubicin, camptothecin, or a combination of the latter two to mimic a dual topoisomerase poison. MLN944 treatment was found to induce a G(1) and G(2) arrest in cells that is unlike the typical G(2)-M arrest noted with known topoisomerase poisons. Finally, transcriptional profiling analysis of xenograft tumors treated with MLN944 revealed clusters of regulated genes distinct from those observed in irinotecan hydrochloride (CPT-11)-treated tumors. Taken together, these findings suggest that the primary mechanism of action of MLN944 likely involves DNA binding and intercalation, but does not appear to involve topoisomerase inhibition. PMID- 14749476 TI - The proteasome inhibitor bortezomib enhances the activity of docetaxel in orthotopic human pancreatic tumor xenografts. AB - Bortezomib (Velcade, formerly known as PS-341) is a boronic acid dipeptide derivative, which is a selective and potent inhibitor of the proteasome. We examined the antitumor activity of combination therapy with bortezomib + docetaxel in two human pancreatic cancer cell lines (MiaPaCa-2 and L3.6pl) selected for their divergent responses to bortezomib alone. Bortezomib blocked docetaxel-induced apoptosis in the MiaPaCa-2 cells and failed to enhance docetaxel-induced apoptosis in L3.6pl cells in vitro but did interact positively with docetaxel to inhibit clonogenic survival. These effects were associated with decreased accumulation of cells in M phase, stabilization of the cyclin-dependent kinase inhibitors, p21 and p27, and inhibition of cdk2 and cdc2 activities. In orthotopic xenografts, combination therapy produced significant reductions in tumor weight and volume in both models associated with accumulation of p21, inhibition of proliferation, and increased apoptosis. Combination therapy also reduced tumor microvessel densities, effects that were associated with reductions in tumor cell production of vascular endothelial growth factor and increased levels of apoptosis in tumor-associated endothelial cells. Together, our results suggest that bortezomib enhances the antitumoral activity of taxanes by enforcing cell growth arrest and inhibiting angiogenesis. PMID- 14749477 TI - Resveratrol modifies the expression of apoptotic regulatory proteins and sensitizes non-Hodgkin's lymphoma and multiple myeloma cell lines to paclitaxel induced apoptosis. AB - Resveratrol (trans-3,4,5-trihydroxystilbene) has received attention for its potential chemopreventive and antitumor effects in experimental systems. Recent evidence suggests that paclitaxel, alone or in combination with other drugs, can be effectively used in the treatment of non-Hodgkin's lymphoma (NHL) and multiple myeloma (MM). This study investigated whether resveratrol can sensitize NHL and MM cell lines to paclitaxel-mediated apoptosis and to delineate the underlying molecular mechanism of sensitization. Both resveratrol and paclitaxel negatively modulated tumor cell growth by arresting the cells at the G(2)-M phase of the cell cycle. Low concentrations of resveratrol exerted a sensitizing effect on drug-refractory NHL and MM cells to apoptosis induced by paclitaxel. Resveratrol selectively down-regulated the expression of antiapoptotic proteins Bcl-x(L) and myeloid cell differentiation factor-1 (Mcl-1) and up-regulated the expression of proapoptotic proteins Bax and apoptosis protease activating factor-1 (Apaf-1). Paclitaxel down-regulated the expression of Bcl-x(L), Mcl-1, and cellular inhibitor of apoptosis protein-1 antiapoptotic proteins and up-regulated Bid and Apaf-1. Combination treatment resulted in apoptosis through the formation of tBid, mitochondrial membrane depolarization, cytosolic release of cytochrome c and Smac/DIABLO, activation of the caspase cascade, and cleavage of poly(adenosine diphosphate-ribose) polymerase. Combination of resveratrol with paclitaxel had minimal cytotoxicity against quiescent and mitogenically stimulated human peripheral blood mononuclear cells. Inhibition of Bcl-x(L) expression by resveratrol was critical for chemosensitization and its functional impairment mimics resveratrol-mediated sensitization to paclitaxel-induced apoptosis. Inhibition of Bcl-x(L) expression by resveratrol was due to the inhibition of the extracellular signal-regulated kinase 1/2 (ERK1/2) pathway and diminished activator protein-1-dependent Bcl-x(L) expression. The findings by resveratrol were corroborated with inhibitors of the ERK1/2 pathway. This study demonstrates that in resistant NHL and MM cell lines resveratrol and paclitaxel selectively modify the expression of regulatory proteins in the apoptotic signaling pathway and the combination, via functional complementation, results in synergistic apoptotic activity. PMID- 14749478 TI - Role of polymorphonuclear leukocytes, nitric oxide synthase, and cyclooxygenase in vascular permeability changes induced by C5a agonist peptides. AB - Tumor responses to radioimmunotherapy combined with peptide agonists of human C5a anaphylatoxin such as GCGYSFKPMPLaR (C5aAP) are two- to four-fold better, depending on the dose of C5aAP, than responses to radioimmunotherapy alone. The enhanced tumor vascular permeability (VP) is the key factor responsible for this improvement. These studies were designed to identify the sequence of events leading to the improved extravasation of immunoglobulin in response to C5aAP. The VP changes were measured in mice after administration of C5aAP alongside of various mediators. The depletion of circulating polymorphonuclear neutrophils (PMN) in mice abolished the C5aAP-induced VP increase. Blocking of P-selectin also returned VP to its basal levels after the C5aAP treatment, indicating that C5aAP-induced VP changes are initiated by interactions of C5aAP with PMNs. Aminoguanidine, an inducible nitric oxide synthase (NOS) inhibitor, given before C5aAP returned VP to control levels. N(omega)-nitro-L-arginine methyl ester, a nonselective NOS inhibitor, had a marginal effect on the activity of C5aAP. Indomethacin, a nonselective cyclooxygenase inhibitor, suppressed C5aAP-induced increases in VP, whereas N-(2-cyclohexyloxy-4-nitrophenyl)-methanesulfonamide, a selective cyclooxygenase-2 inhibitor, was active only at high doses. While C5aAP given i.p. did not alter tumor uptake of (125)I-B72.3, the i.v. administration resulted in approximately 40% increase, confirming the prerequisite interaction of C5aAP with PMNs. The sequence leading to the increased VP appears to be initiated by the interaction of C5aAP with C5a receptor expressed on PMNs followed by binding to endothelial cells of blood vessels. The interaction with P selectin is responsible for the initiation of the nitric oxide cascade as evidenced by inducible NOS activation. Additionally, prostaglandins are required for expression of the full magnitude of the C5aAP activities. PMID- 14749480 TI - Level of glutathione is regulated by ATP-dependent ligation of glutamate and cysteine through photosynthesis in Arabidopsis thaliana: mechanism of strong interaction of light intensity with flowering. AB - Glutathione (GSH) is associated with flowering in Arabidopsis thaliana, but how GSH biosynthesis is regulated to control the transition to flowering remains to be elucidated. Since the key reaction of GSH synthesis is catalyzed by gamma glutamylcysteine synthetase (gamma-ECS) and all the gamma-ECS cDNAs examined contained extra sequences for plastid targeting, we investigated the relationships among GSH levels, photosynthesis and flowering. The GSH level in Arabidopsis increased with the light intensity. The ch1 mutants defective in a light-harvesting antenna in photosystem II showed reduced GSH levels with accumulation of the GSH precursor cysteine, and introduction of the gamma-ECS gene GSH1 under the control of the cauliflower mosaic virus 35S promoter (35S GSH1) into the ch1 mutant altered the GSH level in response to the gamma-ECS mRNA level. These indicate that photosynthesis limits the gamma-ECS reaction to regulate GSH biosynthesis. Like the glutathione-biosynthesis-defective cad2-1 mutant, the ch1 mutants flowered late under weak-light conditions, and this late flowering phenotype was rescued by supplementation of GSH. Introduction of the 35S-GSH1 construct into the ch1 mutant altered flowering in response to the gamma ECS mRNA and GSH levels. These findings indicate that flowering in A. thaliana is regulated by the gamma-ECS reaction of GSH synthesis that is coupled with photosynthesis. PMID- 14749479 TI - Quantitative analysis of p53-targeted gene expression and visualization of p53 transcriptional activity following intratumoral administration of adenoviral p53 in vivo. AB - To analyze the mechanism of the antitumor effect of an adenoviral vector expressing the p53 tumor suppressor (Ad-p53) in vivo, we quantitatively assessed p53-targeted gene expression and visualized transcriptional activity of p53 in tumors in nude mice treated with Ad-p53. Human lung cancer (H1299) xenografts established in nude mice were treated by intratumoral administration of Ad-p53. The levels of expression of exogenous p53 and p53-targeted genes p21, MDM2, Noxa, and p53AIP1 were quantified by real-time reverse transcription-PCR (RT-PCR) and induction of apoptosis was observed histochemically on days 1-3, 7, and 14 after treatment. Expression of mRNA of exogenous p53 and p53-targeted genes (except p53AIP1) was at its maximum 1 day after Ad-p53 treatment and then decreased rapidly; apoptosis was evident in situ 2-3 days after treatment. We developed a noninvasive and simple method for monitoring the transcriptional activity of exogenous p53 following intratumoral administration of Ad-p53 in nude mice. We established H1299 cells that express the green fluorescent protein (GFP) reporter gene under the control of p53-responsive p21 promoter (i.e., the p53R-GFP reporter system). Xenografts of these cells in nude mice were treated by intratumoral administration of Ad-p53, and the transcriptional activity of exogenous p53 could be visualized as intratumoral GFP expression in real time by 3-CCD camera. Expression of GFP was maximal 3 days after treatment and decreased remarkably by 7 days after treatment. We demonstrated that Ad-p53 treatment rapidly induced p53-targeted genes and apoptosis in tumors and succeeded in visualizing p53 transcriptional activity in vivo. We also found that Ad-p53 infection induced phosphorylation of p53 at Ser(46) in p53-sensitive H1299 cells in vitro but not in p53-resistant H226Br cells, suggesting that phosphorylation of Ser(46) is involved in p53-dependent apoptosis. Our data indicate that quantitative analysis of p53-targeted gene expression by real-time quantitative RT-PCR and visualization of p53 transcriptional activity in fresh xenografts by using the p53R-GFP reporter system may be useful in assessing the mechanisms of the antitumor effects of Ad-p53 and novel therapeutic approaches. PMID- 14749481 TI - An ER-localized form of PV72, a seed-specific vacuolar sorting receptor, interferes the transport of an NPIR-containing proteinase in Arabidopsis leaves. AB - Putative vacuolar sorting receptors that bind to the vacuolar targeting signals have been found in various plants; pumpkin PV72, pea BP-80 and Arabidopsis AtELP. PV72 is a seed-specific receptor that is predicted to sort seed storage proteins to protein storage vacuoles. Analysis by surface plasmon resonance showed that the lumenal domain of PV72 bound to an NPIR (a typical vacuolar targeting signal) containing peptide of the precursor of a cysteine proteinase, AtALEU, in the presence of Ca(2+) (K(D) = 0.1 micro M). To elucidate the receptor-dependent transport of vacuolar proteins in plant cells, we produced transgenic Arabidopsis plants that expressed a fusion protein (PV72-HDEL) composed of the lumenal domain of PV72 and an endoplasmic reticulum (ER)-retention signal, HDEL. The expression of PV72-HDEL induced the accumulation of the AtALEU precursor. The accumulation level of the AtALEU precursor was dependent on that of PV72-HDEL. In contrast, it did not induce the accumulation of a precursor of another cysteine proteinase, RD21, which contains no NPIR. Detailed subcellular localization revealed that both the AtALEU precursor and PV72-HDEL accumulated in the ER fraction. We found that most of the AtALEU precursor molecules formed a complex with PV72-HDEL. The AtALEU precursor might be trapped by PV72-HDEL in the ER and not transported to the vacuoles. This in-planta analysis supports the hypothesis that an Arabidopsis homolog of PV72 functions as a sorting receptor for the NPIR-containing proteinase. The overall results suggest that vacuolar sorting receptors for the protein storage vacuoles and the lytic vacuoles share the similar recognition mechanism for a vacuolar targeting signal. PMID- 14749482 TI - Target proteins of the cytosolic thioredoxins in Arabidopsis thaliana. AB - Possible target proteins of cytosolic thioredoxin in higher plants have been investigated in the cell lysate of dark-grown Arabidopsis thaliana whole tissues. We immobilized a mutant of cytosolic thioredoxin, in which an internal cysteine at the active site was substituted with serine, on CNBr activated resin, and used the resin for the thioredoxin-affinity chromatography. By using this resin, the target proteins for thioredoxin in the higher plant cytosol were efficiently acquired. The obtained proteins were separated by two-dimensional gel electrophoresis and analyzed by matrix-assisted laser desorption ionization time of-flight mass spectrometry. Thus we have identified proteins of the anti oxidative stress system proteins (ascorbate peroxidase, germin-like protein, and monomeric type II peroxiredoxin), proteins involved in protein biosynthesis (elongation factor-2 and eukaryotic translation initiation factor 4A), proteins involved in protein degradation (the regulatory subunit of 26S proteasome), and several metabolic enzymes (alcohol dehydrogenase, fructose 1,6-bis phosphate aldolase-like protein, cytosolic glyceraldehyde 3-phosphate dehydrogenase, cytosolic malate dehydrogenase, and vitamin B(12)-independent methionine synthase) together with some chloroplast proteins (chaperonin 60-alpha and 60 beta, heat shock protein 70, and glutamine synthase). The results in this study and recent proteomics studies on the target proteins of chloroplast thioredoxin indicate the versatility and the physiological significance of thioredoxin as reductant in plant cell. PMID- 14749483 TI - Comparative studies on the type-B response regulators revealing their distinctive properties in the His-to-Asp phosphorelay signal transduction of Arabidopsis thaliana. AB - In Arabidopsis thaliana, a Histidine-to-Aspartate (His-->Asp) phosphorelay is involved in the signal transduction for propagation of certain stimuli, such as plant hormones. Through the phosphorelay, the type-B phospho-accepting response regulator (ARR) family members serve as DNA-binding transcriptional regulators, whose activities are most likely regulated by phosphorylation/dephosphorylation. Based on the fact that this higher plant has 11 type-B ARR family genes, we clarified the expression profiles for all of their transcripts in plants. We constructed and characterized a series of transgenic lines, each carrying a given ARR-promoter::GUS transgene. Transcripts of some type-B ARR family genes were detected more or less ubiquitously in many organs tested, while others were expressed predominantly in reproductive organs. These ARR family members were phylogenetically classified into three sub-families, the largest of which includes the well-characterized ARR1, ARR2, and ARR11. Comparative studies were conducted focusing on ARR20 and ARR21, each of which is a representative member of an uncharacterized minor sub-family. A set of transgenic lines was constructed, in each of which a C-terminal DNA-binding domain lacking the N terminal phospho-accepting receiver of a given ARR was aberrantly overexpressed. These resulting transgenic lines, including ARR14-C-ox, ARR20-C-ox, and ARR21-C ox, showed characteristic anomalies during development. These results are discussed with special reference to the His-->Asp phosphorelay signal transduction in A. thaliana. PMID- 14749484 TI - Calcium crystals in the anther of Petunia: the existence and biological significance in the pollination process. AB - Using an X-ray microanalysis system fitted with variable-pressure scanning electron microscopy, we noted that many calcium crystals accumulated under the stomium in the anther of Petunia. When the anther was dehisced and pollen grains were released from the stomata, the calcium crystals adhered to pollen grains and moved to the stigma together with pollen grains. In contrast, an X-ray microanalysis of the stigma surface before pollination detected no calcium emission on the stigma surface. Furthermore, pollen germination and pollen tube growth in medium without Ca occurred as in complete medium. However, after the pollen grains had been washed with abundant germination medium without calcium, pollen germination in the medium without Ca was inhibited. These results show that the calcium crystals dissolved in the aqueous drop under the exudate on the stigma and supplied calcium ions for pollen germination. In addition, calcium crystals were produced not only in the anther of Petunia but also in Nicotiana, suggesting that calcium crystals supply pollen grains with the calcium ions required for pollen germination and serve to improve reproduction efficiency in Solanaceae. PMID- 14749485 TI - Cloning and characterization of glycine-rich RNA-binding protein cDNAs in the moss Physcomitrella patens. AB - We isolated three cDNAs for the genes PpGRP1, PpGRP2 and PpGRP3 that encode glycine-rich RNA-binding proteins (GRPs) from Physcomitrella patens. Three full length cDNA clones were isolated from a cDNA library prepared from poly(A)(+) RNA from 7-day-old protonemata of P. patens. They were named PpGRP1, PpGRP2 and PpGRP3, which encode putative polypeptides of 162, 178 and 155 residues, respectively. Preliminary genomic sequencing suggested that the positions of the three introns in the PpGRP3 gene are similar to those of introns in Arabidopsis GRP genes. PpGRP3 had a putative transit sequence. The PpGRP1-sGFP and PpGRP2 sGFP fusions were targeted to the cell nucleus, while PpGRP3-sGFP fusion was targeted to mitochondria. The level of these PpGRP transcripts as well as that of PpGRP proteins increased after cold treatment. Homoribopolymer RNA assay revealed that PpGRP3 protein show high affinity for poly(U) and poly(G). Results of phylogenetic analysis suggest that the nuclear and mitochondrial forms of GRP have been established early during the evolution of green plants. PMID- 14749486 TI - Characterization of plant circadian rhythms by employing Arabidopsis cultured cells with bioluminescence reporters. AB - Recent intensive studies have begun to shed light on the molecular mechanisms underlying the plant circadian clock in Arabidopsis thaliana. During the course of these previous studies, the most powerful technique, elegantly adopted, was a real-time bioluminescence monitoring system of circadian rhythms in intact plants carrying a luciferase (LUC) fusion transgene. We previously demonstrated that Arabidopsis cultured cells also retain an ability to generate circadian rhythms, at least partly. To further improve the cultured cell system for studies on circadian rhythms, here we adopted a bioluminescence monitoring system by establishing the cell lines carrying appropriate reporter genes, namely, CCA1::LUC and APRR1::LUC, with which CCA1 (CIRCADIAN CLOCK-ASSOCIATED1) and APRR1 (or TOC1) (ARABIDOPSIS PSEUDO-RESPONSE REGULATORS1 or TIMING OF CAB EXPRESSION1) are believed to be the components of the central oscillator. We report the results that consistently supported the view that the established cell lines, equipped with such bioluminescence reporters, might provide us with an advantageous means to characterize the plant circadian clock. PMID- 14749487 TI - Circadian expression of the PpLhcb2 gene encoding a major light-harvesting chlorophyll a/b-binding protein in the moss Physcomitrella patens. AB - Circadian clocks control the expression of Lhcb genes encoding the chlorophyll a/b-binding proteins broadly in seed plants. We show here that this regulation is also conserved in the primitive moss Physcomitrella patens. Northern blotting analyses revealed a robust daily oscillation of Lhcb mRNA levels in protonema cells in 12-h : 12-h light-dark cycles (12 : 12LD) that damped rapidly in continuous darkness (DD). In continuous light (LL), by contrast with typical profiles in higher plants, Lhcb mRNA levels only peaked during the first day and thereafter it showed constant levels. Reverse transcription (RT)-PCR analyses showed similar patterns of expression in LL for three distinct Lhcb genes (PpLhcb1, PpLhcb2 and Zlab1). Moreover, transgenic reporter strains expressing luciferase under the control of the PpLhcb2 promoter showed bioluminescence patterns consistent with the Northern and RT-PCR data. At a higher concentration (4.5%) of glucose in the medium, the reporter strain showed self-sustained rhythms in DD, which was entrained to a differently phased 12 : 12LD, revealing a circadian regulation on the transcription. Kinetics of bioluminescent peaks in 12 : 12LD from gametophore was different to those from protonema, indicating a developmental regulation on PpLhcb2. Together, the regulatory link between the clock and Lhcb genes in P. patens shows characteristics that appear to differ from those in higher plants. PMID- 14749488 TI - Xyloglucan oligosaccharides cause cell wall loosening by enhancing xyloglucan endotransglucosylase/hydrolase activity in azuki bean epicotyls. AB - Addition of xyloglucan-derived oligosaccharides shifted the wall-bound xyloglucans to a lower molecular mass distribution and increased the cell wall extensibility of the native epidermal tissue strips isolated from azuki bean (Vigna angularis) epicotyls. To ascertain the mechanism of oligosaccharide function, we examined the action of a xyloglucan endotransglucosylase/hydrolase (XTH) showing both endotransglucosylase and endohydrolase activities, isolated from azuki bean epicotyl cell walls, in the presence of xyloglucan oligosaccharides. The addition of xyloglucan oligosaccharides enhanced the xyloglucan-degrading activity of XTH against isolated xyloglucan substrates. When the methanol-fixed epidermal tissue strips were incubated with XTH, the molecular mass of wall-bound xyloglucans was decreased and the cell wall extensibility increased markedly in the presence of the oligosaccharides. These results suggest that xyloglucan oligosaccharides stimulate the degradation of xyloglucans by enhancing the XTH activity within the cell wall architecture, thereby increasing the cell wall extensibility in azuki bean epicotyls. PMID- 14749489 TI - Expression and interaction analysis of Arabidopsis Skp1-related genes. AB - Specific protein degradation has been observed in several aspects of development and differentiation in many organisms. One example of such proteolysis is regulated by protein polyubiquitination that is promoted by the SCF complex consisting of Skp1, cullin, and an F-box protein. We examined the activities of the Arabidopsis Skp1-related proteins (ASKs). Among 19 annotated ASK genes, we isolated 16 of the corresponding cDNAs (ASK1, 2, 3, 4, 7, 8, 9, 10, 11, 12, 13, 14, 16, 17, 18, 19), and examined their gene products for interactions with 24 representatives of F-box proteins carrying various classes of the C-terminal domains using the yeast two-hybrid system. As a result, we found diverse binding specificities: ASK1, ASK2, ASK11 and ASK12 interacted well with COI1, FKF1, UFO like protein, LRR-containing F-box proteins, and other F-box proteins with unknown C-terminal motifs. We also observed specific interaction between F-box proteins and ASK3, ASK9, ASK13, ASK14, ASK16 and ASK18. In contrast, we detected no interaction between any of the 12 ASK proteins and F-box proteins containing CRFA, CRFB or CRFC domains. Both histochemical and RT-PCR analysis of eight ASK genes expression revealed unique expression patterns for the respective genes. PMID- 14749490 TI - Significance of the lipid phase in the dynamics and functions of the xanthophyll cycle as revealed by PsbS overexpression in tobacco and in-vitro de-epoxidation in monogalactosyldiacylglycerol micelles. AB - The dynamics of the xanthophyll cycle relative to non-photochemical quenching (NPQ) were examined in tobacco plants overexpressing violaxanthin de-epoxidase (VDE), PsbS and PsbS+VDE for effects on NPQ and violaxanthin (V) de-epoxidation over a range of light intensities. Induction of de-epoxidation and NPQ increased in overexpressed VDE and PsbS plants, respectively. Surprisingly, under low light, overexpressing PsbS enhanced de-epoxidation in addition to NPQ. The effect was hypothesized as due to PsbS binding zeaxanthin (Z) or inducing the binding of Z within the quenching complex, thus shifting the equilibrium toward higher de epoxidation states. Studies in model systems show that Z can stereospecifically inhibit VDE activity against violaxanthin. This effect, observed under conditions of limiting lipid concentration, was interpreted as product feedback inhibition. These results support the hypothesis that the capacity of the thylakoid lipid phase for xanthophylls is limited and modulates xanthophyll-cycle activity, in conjunction with the release of V and binding of Z by pigment-binding proteins. These modulating factors are incorporated into a lipid-matrix model that has elements of a signal transduction system wherein the light-generated protons are the signal, VDE the signal receptor, Z the secondary messenger, the lipid phase the transduction network, and Z-binding proteins the targets. PMID- 14749491 TI - The function of chloroplastic NAD(P)H dehydrogenase in tobacco during chilling stress under low irradiance. AB - The function of chloroplastic NAD(P)H dehydrogenase (NDH) was examined by comparing a tobacco transformant (DeltandhB) in which the ndhB gene had been disrupted with its wild type, upon exposure to chilling temperature (4 degrees C) under low irradiance (100 micro mol m(-2) s(-1) PFD). During the chilling stress, the maximum photochemical efficiency of PSII (F(v)/F(m)) decreased markedly in both the wild type and DeltandhB. However, both F(v)/F(m) and P700(+), as well as the PSII-driven electron transport rate (ETR), in DeltandhB were lower than that in the wild type, implying that NDH-dependent cyclic electron flow around PSI functioned to protect the photosynthetic apparatus from chilling stress under low irradiance. Under the stress, non-photochemical quenching (NPQ), particularly the fast relaxing NPQ component (qf) and the de-epoxidized ratio of the xanthophyll cycle pigments, (A+Z)/(V+A+Z), were distinguishable in DeltandhB from those in the wild type. The lower NPQ in DeltandhB might be related to an inefficient proton gradient across thylakoid membranes (DeltapH) because of lacking an NDH dependent cyclic electron flow around PSI at chilling temperature under low irradiance. PMID- 14749492 TI - An in vivo dual-reporter system of cyanobacteria using two railroad-worm luciferases with different color emissions. AB - In vivo genetic reporter systems using luciferase enzymes enable the real-time monitoring of gene expression in living cells. We have challenged concurrent monitoring of two independent promoter activities within the same cells to precisely compare their characteristics in vivo. In this report, we describe a simple dual-reporter system capable of simultaneously monitoring two promoter activities in living cyanobacterial cells. Two railroad-worm luciferases catalyzing the bioluminescent emissions of different colors served as the dual reporters; each emission was successfully separated by interference filters to estimate the individual bioluminescence signals using photomultiplier tubes. Using this system, we clearly demonstrated the difference in the expression profiles between promoters in the same cells. PMID- 14749493 TI - Functional Shine-Dalgarno-like sequences for translational initiation of chloroplast mRNAs. AB - Many of the chloroplast mRNAs possess Shine-Dalgarno (SD)-like sequences (typically GGAGG) in the 5'-untranslated regions, but the position is highly variable. Using a homologous in vitro translation system, we assessed the role for translation of SD-like sequences in four tobacco chloroplast mRNAs. The rbcL mRNA has a typical SD-like sequence at a position similar to the conserved position (-12 to -4 with respect to the start codon) observed in E. coli, and this sequence was found to be essential for translation. This was also the case for the atpE mRNA. However, SD-like sequences in the rps12 mRNA and in the petB mRNA is located far from (-44 to -42) and too close to (-5 to -2) the initiation codon, respectively, and these sequences were not essential for translation. These results indicate that functional SD-like sequences are located around 10 nucleotides upstream from the translational start codon. Competition assays confirmed that a functional SD-like sequence interacts with the 3' terminus of chloroplast 16S rRNA. PMID- 14749494 TI - Essential roles of Her2/erbB2 in cardiac development and function. AB - The tyrosine kinase receptor erbB2, also known in humans as Her2, is a member of the epidermal growth factor receptor (EGFR or erbB1) family, which also includes erbB3 and erbB4. The erbBs were discovered in an avian erythroblastosis tumor virus and exhibited similarities to human EGFR (Yarden and Sliwkowski, 2001). Her2/erbB2 is highly expressed in many cancer types. Its overexpression is correlated with a poor prognosis for breast and ovarian cancer patients. ErbB receptors bind to a family of growth factors, termed neuregulins/heregulin (NRG/HRG), which comprise NRG-1, -2, -3, and -4 and include multiple isoforms. ErbB2/Her2 is an orphan receptor that does not bind ligand alone but heterodimerizes with the other erbB receptors for NRG signaling. ErbB2 is expressed in multiple neuronal and non-neuronal tissues in embryos and adult animals, including the heart. Genetic data demonstrated that erbB2 is required for normal embryonic development of neural crest-derived cranial sensory neurons. ErbB2/Her2-null mutant embryos of a trabeculation defect die before embryonic day (E) 11. To study its role at later stages of development, we generated a transgenic mouse line that specifically expresses the rat erbB2 cDNA in the heart under the control of the cardiac-specific alpha-myosin heavy chain promoter. When crossed into the null background, the expression of the rat erbB2 cDNA rescued the cardiac phenotype in the erbB2-null mutant mice that survive until birth but display an absence of Schwann cells and a severe loss of both motor and spinal sensory neurons. To study the role of erbB2 in the adult heart, we generated conditional mutant mice carrying a cardiac-restricted deletion of erbB2. These erbB2 conditional mutants exhibited multiple independent parameters of dilated cardiomyopathy, including chamber dilation, wall thinning, and decreased contractility. Interestingly, treatment of breast cancers overexpressing erbB2 with Herceptin (Trastuzumab), a humanized monoclonal antibody specific to the extracellular domain of erbB2, results in some patients developing cardiac dysfunction. The adverse effect is increased significantly in those patients who also receive the chemotherapeutical agent anthracycline. We found that erbB2 deficient cardiac myocytes are more susceptible to anthracycline-induced cytotoxicity. These results suggest that erbB2 signaling in the heart is essential for the prevention of dilated cardiomyopathy. These lines of mice provide models with which to elucidate the molecular and cellular mechanisms by which erbB2 signaling regulates cardiac functions. These mice also will provide important information for devising strategies to mitigate the cardiotoxic effects of Herceptin treatment, allowing for the potential expanded use of this drug to treat all cancers overexpressing erbB2. PMID- 14749495 TI - The adrenergic pathway and heart failure. AB - Heart failure represents the endpoint to many triggering cardiovascular pathologies. However, there are molecular and biochemical features that remain common to the failing heart, despite the varying etiologies. Principal among these is heightened activation of the sympathetic nervous system and associated enhancement of adrenergic signaling pathways via the catecholamines, norepinephrine and epinephrine. During heart failure, several hallmark alterations in the adrenergic system contribute to loss of cardiac function. To specifically study these changes in a physiologically relevant setting, we and others have utilized advances in genetically engineered mouse technology. This chapter will discuss the many transgenic and knockout mouse models that have been developed to study the adrenergic system in the normal and failing heart. These models include genetically manipulated alterations of adrenergic receptors, linked heterotrimeric G proteins, and the regulatory G protein-coupled receptor kinases (GRKs). Among the more-interesting information gained from these models is the finding that inhibition of a particular GRK - GRK2 or beta adrenergic receptor kinase 1 (betaARK1) - is a potential novel therapeutic strategy to improve function in the setting of heart failure. Furthermore, we will discuss recent transgenic research that proposes an important role for hypertension in the development of heart failure. Overall, genetically engineered mouse models pertaining to this critical myocardial signaling system have provided novel insight into heart function under normal conditions and during states of dysfunction and failure. PMID- 14749496 TI - Effects of thyroid hormone on the cardiovascular system. AB - Increased or reduced action of thyroid hormone on certain molecular pathways in the heart and vasculature causes relevant cardiovascular derangements. It is well established that overt hyperthyroidism induces a hyperdynamic cardiovascular state (high cardiac output with low systemic vascular resistance), which is associated with a faster heart rate, enhanced left ventricular (LV) systolic and diastolic function, and increased prevalence of supraventricular tachyarrhythmias - namely, atrial fibrillation - whereas overt hypothyroidism is characterized by the opposite changes. However, whether changes in cardiac performance associated with overt thyroid dysfunction are due mainly to alterations of myocardial contractility or to loading conditions remains unclear. Extensive evidence indicates that the cardiovascular system responds to the minimal but persistent changes in circulating thyroid hormone levels, which are typical of individuals with subclinical thyroid dysfunction. Subclinical hyperthyroidism is associated with increased heart rate, atrial arrhythmias, increased LV mass, impaired ventricular relaxation, reduced exercise performance, and increased risk of cardiovascular mortality. Subclinical hypothyroidism is associated with impaired LV diastolic function and subtle systolic dysfunction and an enhanced risk for atherosclerosis and myocardial infarction. Because all cardiovascular abnormalities are reversed by restoration of euthyroidism ("subclinical hypothyroidism") or blunted by beta-blockade and L-thyroxine (L-T4) dose tailoring ("subclinical hyperthyroidism"), timely treatment is advisable in an attempt to avoid adverse cardiovascular effects. Interestingly, some data indicate that patients with acute and chronic cardiovascular disorders and those undergoing cardiac surgery may have altered peripheral thyroid hormone metabolism that, in turn, may contribute to altered cardiac function. Preliminary clinical investigations suggest that administration of thyroid hormone or its analogue 3,5 diiodothyropropionic acid greatly benefits these patients, highlighting the potential role of thyroid hormone treatment in patients with acute and chronic cardiovascular disease. PMID- 14749497 TI - Functional significance of Tie2 signaling in the adult vasculature. AB - Abundant data now demonstrate that the growth of new blood vessels, termed angiogenesis, plays both pathological and beneficial roles in human disease. Based on these data, a tremendous effort has been undertaken to understand the molecular mechanisms that drive blood vessel growth in adult tissues. Tie2 recently was identified as a receptor tyrosine kinase expressed principally on vascular endothelium. Disrupting Tie2 function in mice resulted in embryonic lethality with defects in embryonic vasculature, suggesting a role in blood vessel maturation and maintenance. Based on these studies, we undertook a series of studies to probe the function of Tie2 in adult vasculature that will form the focus of this chapter. Consistent with a role in blood vessel growth in adult vasculature, Tie2 was upregulated and activated in the endothelium of rat ovary and in healing rat skin wounds, both areas of active angiogenesis. Moreover, Tie2 was upregulated in the endothelium of vascular "hot spots" in human breast cancer specimens. Surprisingly, Tie2 also was expressed and activated in the endothelium of all normal rat tissues examined, suggesting a role in maintenance of adult vasculature. To determine the functional role of Tie2 in tumor vasculature, a soluble Tie2 extracellular domain (ExTek) was designed that blocked the activation of Tie2 by its activating ligand, angiopoietin 1 (Ang1). Administration of recombinant ExTek protein or an ExTek adenovirus inhibited tumor growth and metastasis in rodent tumor models, demonstrating a functional role for Tie2 in pathological angiogenesis in adult tissues. To begin to understand the endothelial signaling pathways and cellular responses that mediate Tie2 function, we identified signaling molecules that are recruited to the activated, autophosphorylated Tie2 kinase domain. Two of these molecules, SHP2 and GRB2, are part of the pathway upstream of mitogen-activated protein kinase (MAPK) activation, a pathway that may be responsible for morphogenetic effects of Tie2 on endothelial cells. Another signaling molecule, p85, is responsible for recruitment of phosphatidylinositol 3 kinase (PI3-K) and activation of the Akt/PI3-K pathway. Akt/PI3-K has emerged as a critical pathway downstream of Tie2 that is necessary for cell survival effects as well as for chemotaxis, activation of endothelial nitric oxide synthase, and perhaps for anti-inflammatory effects of Tie2 activation. Taken together, these studies and many others demonstrate that the Tie2 pathway has important functions in adult tissues, in both quiescent vasculature and during angiogenesis, and help to validate the Tie2 pathway as a therapeutic target. PMID- 14749498 TI - Angiostatin and anti-angiogenic therapy in human disease. AB - Many diseases have abnormal quality and/or quantity of vascularization as a characteristic feature. Cancer cells elicit the growth of new capillaries during neovascularization in a process termed angiogenesis. In diabetics, pathologic angiogenesis in various tissues is a clinical feature of many common complications. Therefore, the diabetic cancer patient warrants special consideration and extra care in the design of anti-angiogenic treatments without adverse side effects. Some treatment regimens that look promising in vitro, in animal models, or in early clinical trials may be contra-indicated for diabetics. This chapter will review the common complications of diabetes, with emphasis on the angiogenic pathology. Recent research related to the mechanism of action and basis for specificity of the anti-angiogenic peptide, angiostatin, will be the focus. The aim is to shed light on areas in which more research is needed with respect to angiostatin and other anti-angiogenic agents and the microenvironmental conditions that affect their activities, in order to develop improved therapeutic strategies for diabetic cancer patients. PMID- 14749499 TI - Balancing contractility and energy production: the role of myocyte enhancer factor 2 (MEF2) in cardiac hypertrophy. AB - Cardiac hypertrophy -- that is, enlargement of the heart resulting from increased myocyte size -- is observed with many forms of human heart disease. It may arise secondary to an insult, such as infarct or chronic hypertension, or may occur as a consequence of a genetic defect, such as in hypertrophic cardiomyopathy. Traditionally, it has been widely believed that hypertrophy occurred as an adaptive response to normalize increased wall stress due to disease. Recently, however, it has been observed that while hypertrophy initially appears to improve the function of the heart following insult, over time, it frequently leads to a decompensated state, characterized by fibrosis and chamber dilation, resulting in overt heart failure. Hypertrophy also occurs during fetal development, immediately after birth, and in trained athletes; however, it does not lead to decompensation in these states. Experiments over the last 15 years have implicated similar signaling pathways in both pathological and physiological hypertrophic responses. Recently, important differences have been demonstrated that might hold the key to the development of effective new treatments for human diseases. This chapter focuses on how these hypertrophic responses differ from one another phenotypically and discusses how inefficient or impaired energy metabolism in the heart may contribute to the development of pathological responses. We also discuss recent evidence that the myocyte enhancer factor 2 (MEF2) transcription factor family, which previously has been shown to be important in cardiac development and hypertrophy, may have a role in regulation of cardiac energy metabolism. PMID- 14749500 TI - Phosphorylation of RNA polymerase II in cardiac hypertrophy: cell enlargement signals converge on cyclin T/Cdk9. AB - Cardiac myocyte enlargement is the eponymous characteristic of cardiac hypertrophy, regardless of the instigating signal. Such triggers include biomechanical stress (e.g., work load, compensation for ischemic damage), sarcomeric protein mutations, cytoskeletal protein mutations, abnormal energetics, G protein-coupled receptors for ligands (including angiotensin II and endothelin-1), or their signal transducers within cells. In turn, increased myocyte size reflects increased RNA and protein content per cell as responses to these stimuli. In eukaryotic cells, the large subunit of RNA polymerase II (RNAPII) becomes extensively phosphorylated in its serine-rich C-terminal domain (CTD) during the transition from transcript initiation to transcript elongation - that is, "escape" of RNAPII from the promoter-proximal region into the open reading frame. Although this process is believed to be crucial to productive synthesis of mRNA and is known to be governed by two atypical cyclin-dependent kinases, Cdk7 and Cdk9, surprisingly little is understood of how regulatory pathways within cells intersect these RNAPII-directed protein kinases. Investigations of the CTD kinase module in cardiac hypertrophy provide a tentative initial map of a molecular circuit controlling cell size through regulated phosphorylation of RNAPII. PMID- 14749501 TI - Cardiomyocyte calcium and calcium/calmodulin-dependent protein kinase II: friends or foes? AB - Calcium (Ca(2+)) is a critical second messenger in cell signaling. Elevated intracellular Ca(2+) can activate numerous Ca(2+)-regulated enzymes. These enzymes have different subcellular localizations and may respond to distinct modes of Ca(2+) mobilization. In cardiac muscle, Ca(2+) plays a central role in regulating contractility, gene expression, hypertrophy, and apoptosis. Many cellular responses to Ca(2+) signals are mediated by Ca(2+)/calmodulin-dependent enzymes, among which is the Ca(2+)/calmodulin-dependent protein kinase II (CaMKII). Putative substrates for CaMKII include proteins involved in regulating Ca(2+) storage and release, transcription factors, and ion channels. The major isoform of CaMKII in the heart is CaMKIIdelta. Two cardiac splice variants, CaMKIIdelta(B) and delta(C), differ in whether they contain a nuclear localization sequence. Our laboratory has examined the hypothesis that the nuclear delta(B) and the cytoplasmic delta(C) isoforms respond to different Ca(2+) stimuli and have distinct effects on hypertrophic cardiac growth and Ca(2+) handling. We have shown that pressure overload-induced hypertrophy differentially affects the nuclear delta(B) and the cytoplasmic delta(C) isoforms of CaMKII. Additionally, using isolated myocytes and transgenic mouse models, we demonstrated that the nuclear CaMKIIdelta(B) isoform plays a key role in cardiac gene expression associated with cardiac hypertrophy. The cytoplasmic CaMKIIdelta(C) isoform phosphorylates substrates involved in Ca(2+) handling. Dysregulation of intracellular Ca(2+) and resulting changes in excitation contraction coupling characterize heart failure and can be induced by in vivo overexpression of CaMKIIdelta(C) and phosphorylation of its substrates. The differential location of CaMKII isoforms and their relative activation by physiological vs. pathological stimuli may provide a paradigm for exploring and elucidating how Ca(2+)/CaMKII pathways can serve as both friends and foes in the heart. PMID- 14749502 TI - Hypertension and obesity. AB - Obesity is a common problem in much of the western world today in that is linked directly with several disease processes, notably, hypertension. It is becoming clear that the adipocyte is not merely an inert organ for storage of energy but that it also secretes a host of factors that interact with each other and may result in elevated blood pressure. Of particular importance is the putative role of leptin in the causation of hypertension via an activation of the sympathetic nervous system and a direct effect on the kidneys, resulting in increased sodium reabsorption leading to hypertension. Obesity per se may have structural effects on the kidneys that may perpetuate hypertension, leading to an increased incidence of end-stage renal disease that results in further hypertension. Adipose tissue may elaborate angiotensin from its own local renin-angiotensin system. The distribution of body fat is considered important in the genesis of the obesity-hypertension syndrome, with a predominantly central distribution being particularly ominous. Weight loss is the cornerstone in the management of the obesity-hypertension syndrome. It may be achieved with diet, exercise, medications, and a combination of these measures. Anti-obesity medications that are currently undergoing clinical trials may play a promising role in the management of obesity and may also result in lowering of blood pressure. Antihypertensives are considered important components in the holistic approach to the management of this complex problem. PMID- 14749503 TI - Obesity, insulin resistance, and cardiovascular disease. AB - The ability of insulin to stimulate glucose disposal varies more than six-fold in apparently healthy individuals. The one third of the population that is most insulin resistant is at greatly increased risk to develop cardiovascular disease (CVD), type 2 diabetes, hypertension, stroke, nonalcoholic fatty liver disease, polycystic ovary disease, and certain forms of cancer. Between 25-35% of the variability in insulin action is related to being overweight. The importance of the adverse effects of excess adiposity is apparent in light of the evidence that more than half of the adult population in the United States is classified as being overweight/obese, as defined by a body mass index greater than 25.0 kg/m(2). The current epidemic of overweight/obesity is most-likely related to a combination of increased caloric intake and decreased energy expenditure. In either instance, the fact that CVD risk is increased as individuals gain weight emphasizes the gravity of the health care dilemma posed by the explosive increase in the prevalence of overweight/obesity in the population at large. Given the enormity of the problem, it is necessary to differentiate between the CVD risk related to obesity per se, as distinct from the fact that the prevalence of insulin resistance and compensatory hyperinsulinemia are increased in overweight/obese individuals. Although the majority of individuals in the general population that can be considered insulin resistant are also overweight/obese, not all overweight/obese persons are insulin resistant. Furthermore, the cluster of abnormalities associated with insulin resistance - namely, glucose intolerance, hyperinsulinemia, dyslipidemia, and elevated plasma C-reactive protein concentrations -- is limited to the subset of overweight/obese individuals that are also insulin resistant. Of greater clinical relevance is the fact that significant improvement in these metabolic abnormalities following weight loss is seen only in the subset of overweight/obese individuals that are also insulin resistant. In view of the large number of overweight/obese subjects at potential risk to be insulin resistant/hyperinsulinemic (and at increased CVD risk), and the difficulty in achieving weight loss, it seems essential to identify those overweight/obese individuals who are also insulin resistant and will benefit the most from weight loss, then target this population for the most intensive efforts to bring about weight loss. PMID- 14749504 TI - Leptin and the cardiovascular system. AB - Obesity is associated with increased cardiovascular morbidity and mortality, in part through development of hypertension. Recent observations suggest that the cardiovascular actions of leptin may help explain the link between excess fat mass and cardiovascular diseases. Leptin is an adipocyte-derived hormone that acts in the central nervous system to promote weight loss by decreasing food intake and increasing metabolic rate. Leptin causes a significant increase in overall sympathetic nervous activity, which appears to be due to direct hypothalamic effects and is mediated by neuropeptide systems such as the melanocortin system and corticotropin-releasing hormone. Renal sympathoactivation to leptin is preserved in the presence of obesity, despite resistance to the metabolic effects of leptin. Such selective leptin resistance, in the context of circulating hyperleptinemia, could predispose to obesity-related hypertension. Some in vitro studies have suggested that leptin may have peripheral actions such as endothelium-mediated vasodilation that might oppose sympathetically induced vasoconstriction. However, we and others have shown that leptin does not have direct vasodilator effects in vivo. The fact that chronic leptin administration or overexpression of leptin produces hypertension supports the concept that the hemodynamic actions of leptin are due predominantly to sympathetic activation. Exploration of the sites and mechanisms of leptin resistance should provide novel therapeutic strategies for obesity, insulin resistance, and hypertension. PMID- 14749505 TI - The use of animal models to dissect the biology of leptin. AB - Our understanding of the effects of leptin have stemmed mainly from animal studies, which continue to leave important clues of its roles in physiology, metabolism, neuroscience, and cell signaling. Since its discovery, leptin has been linked to various pathways, either directly at its primary site of action in the hypothalamus, or indirectly via downstream effector pathways such as in adipocytes and muscle. Leptin's importance is exemplified by the lack of redundant backup mechanisms, since leptin-deficient mice are obese, diabetic, and sterile. Investigations uncovering the pleiotropic actions of leptin were unfolded mainly from rodent models. Thus, this chapter focuses on these studies and, more specifically, on those findings recently brought forward by transgenic mice overexpressing leptin. The vast amount of biology that has been ascribed to leptin encompasses effects on food intake, insulin sensitivity, adiposity, thermogenesis, reproduction, immunity, and bone regulation. Mechanisms underlying leptin's action revolve essentially around neural pathways but also encompass to a lesser extent peripheral mechanisms. The roles of leptin along these axes are reviewed, with particular emphasis on pathways and phenotypes generated by transgenic hyperleptinemia. An evolutionary significance of hyperleptinemia in association with development of leptin resistance is suggested as a protective armament against some of the detrimental effects caused by hyperleptinemia in transgenic mice overexpressing leptin. PMID- 14749506 TI - Insulin and leptin as adiposity signals. AB - There is now considerable consensus that the adipocyte hormone leptin and the pancreatic hormone insulin are important regulators of food intake and energy balance. Leptin and insulin fulfill many of the requirements to be putative adiposity signals to the brain. Plasma leptin and insulin levels are positively correlated with body weight and with adipose mass in particular. Furthermore, both leptin and insulin enter the brain from the plasma. The brain expresses both insulin and leptin receptors in areas important in the control of food intake and energy balance. Consistent with their roles as adiposity signals, exogenous leptin and insulin both reduce food intake when administered locally into the brain in a number of species under different experimental paradigms. Additionally, central administration of insulin antibodies increases food intake and body weight. Recent studies have demonstrated that both insulin and leptin have additive effects when administered simultaneously. Finally, we recently have demonstrated that leptin and insulin share downstream neuropeptide signaling pathways. Hence, insulin and leptin provide important negative feedback signals to the central nervous system, proportional to peripheral energy stores and coupled with catabolic circuits. PMID- 14749507 TI - Leptin receptor signaling and the regulation of mammalian physiology. AB - While the hormone leptin and its receptor were discovered relatively recently, a great deal is already known about the molecular details of leptin receptor (LR) signaling and physiologic regulation. While multiple alternatively spliced LR isoforms exist, only the long (LRb) form associates with the Janus kinase 2 (Jak2) tyrosine kinase to mediate intracellular signaling. LRb initiates signaling via three major mechanisms: 1) Tyr(985) of LRb recruits SH2-containing tyrosine phosphatase (SHP-2); 2) Tyr(1138) of LRb recruits signal transducer and activator of transcription 3 (STAT3); and 3) tyrosine phosphorylation sites on the receptor-associated Jak2 likely recruit numerous undefined signaling proteins. The Tyr(985) --> SHP-2 pathway is a major regulator of extracellular signal-regulated kinase (ERK) activation during leptin signaling in cultured cells, while the Tyr(1138) --> STAT3 pathway induces the feedback inhibitor, suppressor of cytokine signaling 3 (SOCS3), as well as important positive effectors of leptin action. The Jak2-dependent activation of the insulin receptor substrate (IRS) protein --> phosphatidylinositol 3-kinase (PI3'-K) pathway appears to regulate membrane potential in LRb-expressing neurons and contributes to the regulation of feeding. The Tyr(1138) --> STAT3 pathway mediates transcriptional regulation of the hypothalamic melanocortin pathway in vivo. This pathway is required for the regulation of appetite and energy expenditure by leptin. Interestingly, the Tyr(1138) --> STAT3 pathway does not strongly regulate neuropeptide Y (NPY) and thus is not required for the control of reproduction and growth. Thus, other as-yet-undefined leptin receptor signals are central to these and perhaps other aspects of leptin action. PMID- 14749508 TI - Leptin signaling in the central nervous system and the periphery. AB - The discovery of leptin in 1994 has led to astonishing advances in understanding the regulation of energy balance in rodents and humans. The demonstration of leptin receptors in hypothalamic regions known to play critical roles in regulating energy intake and body weight has produced considerable excitement in the field. Most attention has focused on the central actions of leptin. The receptor is present in several populations of neurons that express specific appetite-regulating neuropeptides for which both expression and release are regulated by leptin. Recent advances show that central leptin action is not limited to influencing energy balance. Leptin regulates a broad variety of processes and behaviors, such as blood pressure, neuroendocrine axes, bone mass, and immune function. The cloning of leptin receptors also led to parallel studies examining their signaling capacities in mammalian cell lines. The long-form receptor regulates multiple intracellular signaling cascades, including the classic janus activating kinase-signal transducer and activator of transcription (JAK-STAT) pathway, consistent with belonging to the cytokine-receptor superfamily and the phosphoinositol-3 kinase and adenosine monophosphate kinase pathways. Progress has been made in understanding the role of individual signaling pathways in vivo and the mechanisms by which specific neuropeptides are regulated. Regulation of the pro-opiomelanocortin (pomc) and the thyrotropin releasing hormone (trh) genes by leptin is particularly well understood. Novel players in negative regulation of central leptin receptor signaling have been identified and open the possibility that these may be important in the development of leptin resistance and obesity. While initial focus was on the central effects of leptin, important actions have been discovered in peripheral tissues. These include roles of leptin to directly regulate immune cells, pancreatic beta cells, adipocytes, and muscle cells. Recent elucidation of a new signaling pathway in skeletal muscle affecting fatty acid metabolism has implications for regulation of insulin sensitivity and glucose metabolism. Recent progress in understanding central and peripheral leptin receptor signaling provides potential new targets for anti-obesity and anti-diabetes drug development. PMID- 14749509 TI - Polymorphisms in the glucocorticoid receptor gene and their associations with metabolic parameters and body composition. AB - Most actions of glucocorticoids (GCs) are mediated by the glucocorticoid receptor (GR). The interindividual response to GCs varies considerably, as demonstrated by a variable suppressive response to 0.25-mg dexamethasone (DEX). Several polymorphisms in the gene coding for the GR have been described. It is unclear to what extent the observed response variability is due to GR polymorphisms or to other factors. However, at least three polymorphisms seem to be associated with altered GC sensitivity and changes in body composition and metabolic parameters. The N363S polymorphism has been associated with increased sensitivity to GCs, increased insulin response to DEX, a tendency towards lower bone mineral density, and increased body mass index (BMI). However, other reports found no associations with BMI. Another polymorphism, previously described as a BclI restriction fragment length polymorphism, recently was identified as a C --> G nucleotide change. The G allele also was associated with increased sensitivity to GCs. In middle-aged subjects, the G allele of this BclI polymorphism was associated with increased abdominal obesity, while at older age, a lower BMI was found, accompanied by a tendency towards lower lean body mass. A third polymorphism consists of two linked, single-nucleotide mutations in codons 22 and 23, of which the second mutation results in an amino acid change from arginine (R) to lysine (K). In contrast to the other polymorphisms, this ER22/23EK polymorphism was associated with a relative resistance to GCs. In line with this, ER22/23EK carriers had lower total cholesterol and low-density lipoprotein cholesterol levels as well as lower fasting insulin concentrations and a better insulin sensitivity. C-reactive protein levels were lower in ER22/23EK carriers, as was found in a different population of elderly males. In accordance with this healthy metabolic profile, we found in this population a significantly better survival in ER22/23EK carriers after a 4-year follow-up. GCs also affect the brain. Although a certain level of cortisol is essential for proper brain functioning, excessive GC levels have been shown to negatively affect brain morphology and functions. At older age, we found that the risk of dementia and white matter lesions was lower in ER22/23EK carriers. GCs are also important in the regulation of body fat distribution. At young age, we observed sex-specific differences in body composition. Male ER22/23EK carriers were taller, had more muscle mass, and were stronger than noncarriers. In young females, ER22/23EK carriers had tendencies towards smaller waist and hip circumferences and lower body weight. Another polymorphism (TthIIII) was not associated with altered GC sensitivity. In conclusion, these polymorphisms in the GR gene may contribute considerably to the observed variability in GC sensitivity. As a result, they are associated with several differences in body composition and metabolic factors. PMID- 14749510 TI - Glucocorticoids and 11beta-hydroxysteroid dehydrogenase in adipose tissue. AB - The highly prevalent metabolic syndrome (insulin resistance, type 2 diabetes, dyslipidemia, hypertension, along with abdominal obesity) resembles Cushing's syndrome. However, in simple obesity, plasma cortisol levels are not elevated. 11beta-hydroxysteroid dehydrogenase type 1 (11beta-HSD1), at least in mature adipocytes and hepatocytes, converts inactive circulating 11-keto steroids into active glucocorticoids, amplifying local glucocorticoid action. 11beta-HSD1 is elevated in adipose tissue in obese humans and rodents, suggesting that adipose tissue glucocorticoid excess may explain the conundrum. Indeed, transgenic mice overexpressing 11beta-HSD1 in adipose tissue faithfully replicate the metabolic syndrome. Conversely, 11beta-HSD1(-/-) mice resist the metabolic consequences of stress and high-fat feeding via insulin sensitisation and other advantageous effects in the liver and adipose tissue. Adipose 11beta-HSD1 deficiency contributes to a protective metabolic phenotype, supporting its role as a therapeutic target for the metabolic syndrome. PMID- 14749511 TI - The central melanocortin system and the integration of short- and long-term regulators of energy homeostasis. AB - The importance of the central melanocortin system in the regulation of energy balance is highlighted by studies in transgenic animals and humans with defects in this system. Mice that are engineered to be deficient for the melanocortin-4 receptor (MC4R) or pro-opiomelanocortin (POMC) and those that overexpress agouti or agouti-related protein (AgRP) all have a characteristic obese phenotype typified by hyperphagia, increased linear growth, and metabolic defects. Similar attributes are seen in humans with haploinsufficiency of the MC4R. The central melanocortin system modulates energy homeostasis through the actions of the agonist, alpha-melanocyte-stimulating hormone (alpha-MSH), a POMC cleavage product, and the endogenous antagonist AgRP on the MC3R and MC4R. POMC is expressed at only two locations in the brain: the arcuate nucleus of the hypothalamus (ARC) and the nucleus of the tractus solitarius (NTS) of the brainstem. This chapter will discuss these two populations of POMC neurons and their contribution to energy homeostasis. We will examine the involvement of the central melanocortin system in the incorporation of information from the adipostatic hormone leptin and acute hunger and satiety factors such as peptide YY (PYY(3-36)) and ghrelin via a neuronal network involving POMC/cocaine and amphetamine-related transcript (CART) and neuropeptide Y (NPY)/AgRP neurons. We will discuss evidence for the existence of a similar network of neurons in the NTS and propose a model by which this information from the ARC and NTS centers may be integrated directly or via adipostatic centers such as the paraventricular nucleus of the hypothalamus (PVH). PMID- 14749512 TI - Monogenic human obesity syndromes. AB - Over the past decade, we have witnessed a major increase in the scale of scientific activity devoted to the study of energy balance and obesity. This explosion of interest has, to a large extent, been driven by the identification of genes responsible for murine obesity syndromes and the novel physiological pathways revealed by those genetic discoveries. We and others recently have identified several single-gene defects causing severe human obesity. Many of these defects have occurred in molecules identical or similar to those identified as a cause of obesity in rodents. This chapter will consider the human monogenic obesity syndromes that have been characterized to date and discuss how far such observations support the physiological role of these molecules in the regulation of human body weight and neuroendocrine function. PMID- 14749513 TI - Effect of routine MR imaging of the brain at 1.5 T on subsequent magnetoencephalography: results in nine volunteers. AB - PURPOSE: To investigate whether routine magnetic resonance (MR) imaging of the brain with a whole-body 1.5-T imager affects the results of subsequent magnetoencephalography (MEG). MATERIALS AND METHODS: Nine healthy volunteers (six women, mean age of 23 years, age range of 20-27 years; three men, mean age of 24 years, age range of 23-25 years) underwent one MEG session before and two MEG sessions after MR imaging of the brain. The first MEG session was completed about 20 minutes before brain MR imaging began, the second MEG session (MEG 2) was performed within 30 minutes after MR imaging, and the third MEG session was performed 2 hours after MEG 2. Each MEG session involved measurement of spontaneous brain activity and, in seven patients, of brain activity during stimulation of the median nerve. MR imaging included T1- and T2-weighted fast spin-echo and gradient-echo sequences applied with a 1.5-T clinical imager. Data were compared by using a repeated-measures analysis of variance (general linear model) both with and without Greenhouse-Geisser correction. RESULTS: MEG signals were detected and measured without difficulty in all volunteers. No statistically significant difference was seen between estimated noise at MEG before and after MR imaging (P =.588 with correction, P =.665 without correction). MEG records obtained in all volunteers enabled localization of evoked response to median nerve stimulation before and after MR imaging. No measurable differences were observed between relative power spectra of spontaneous brain activity before and after MR imaging (P >.290 with correction, P >or=.295 without correction). CONCLUSION: No measurable effect of 1.5-T brain MR imaging on subsequent MEG was detected. PMID- 14749514 TI - Primary vertebral osteosarcoma: imaging findings. AB - PURPOSE: To evaluate patient age and sex and location and imaging appearances of primary vertebral osteosarcoma (PVOS) compared with histologic subtypes. MATERIALS AND METHODS: Retrospective review (1915-2001) of imaging findings in patients with histologically proved primary osteosarcoma of vertebral column was performed. Two radiologists in consensus reviewed findings for location, origin site, matrix pattern, and spinal canal invasion and compared them with histologic subtypes. Radiation-induced, Paget, metastatic, and multifocal osteosarcoma were excluded. RESULTS: Of 4,887 osteosarcoma cases, 198 (4%) were PVOS arising from vertebral column. There were 103 female and 95 male patients (age range, 8-80 years; median age, 34.5 years). Involvement included cervical (27 patients), thoracic (66 patients), lumbar (64 patients), and sacral (41 patients) spine. Adequate imaging findings were available in 69 patients, and involvement of two levels was seen in 12 (17%). In nonsacral spine, most tumors (44 cases) arose from posterior elements, with partial involvement of vertebral body. Lesions confined to vertebral body were less frequent (12 cases). Sacral tumors involved body and sacral ala. The most common histologic subtypes were osteoblastic (47 patients), chondroblastic (12 patients), telangiectatic (four patients), fibroblastic (four patients), small cell (one patient), and epithelioid (one patient). The majority (55 cases) demonstrated osteoid matrix mineralization; 17 showed marked mineralization. Five cases with marked mineralization were confined to vertebral body, with "ivory vertebra" appearance. Purely lytic pattern was seen in 14 (20%) cases. Lytic pattern was seen in four (100%) telangiectatic, three (75%) fibroblastic, three (25%) chondroblastic, three (6%) conventional osteoblastic, and one (100%) small-cell subtypes. Invasion of spinal canal was common (84% of cases). Appearance simulating osteoblastoma without soft-tissue mass was present (seven cases). Pathologic compression fractures were identified (seven patients). CONCLUSION: This study provides age and sex distribution and location and imaging features in a large series of PVOS. PMID- 14749515 TI - NMR structure of the KaiC-interacting C-terminal domain of KaiA, a circadian clock protein: implications for KaiA-KaiC interaction. AB - KaiA is a two-domain circadian clock protein in cyanobacteria, acting as the positive element in a feedback loop that sustains the oscillation. The structure of the N-terminal domain of KaiA is that of a pseudo-receiver, similar to those of bacterial response regulators, which likely interacts with components of the clock-resetting pathway. The C-terminal domain of KaiA is highly conserved among cyanobacteria and enhances the autokinase activity of KaiC. Here we present the NMR structure of the C-terminal domain of KaiA from the thermophilic cyanobacterium Thermosynechococcus elongatus BP-1. This domain adopts a novel all alpha-helical homodimeric structure. Several mutations known to affect the period of the circadian oscillator are shown to be located at an exposed groove near the dimer interface. This NMR structure and a 21-A-resolution electron microscopy structure of the hexameric KaiC particle allow us to postulate a mode of KaiA KaiC interaction, in which KaiA binds a linker region connecting two globular KaiC domains. PMID- 14749516 TI - Airborne signals prime plants against insect herbivore attack. AB - Green leafy volatiles (GLV), six-carbon aldehydes, alcohols, and esters commonly emitted by plants in response to mechanical damage or herbivory, induced intact undamaged corn seedlings to rapidly produce jasmonic acid (JA) and emit sesquiterpenes. More importantly, corn seedlings previously exposed to GLV from neighboring plants produced significantly more JA and volatile sesquiterpenes when mechanically damaged and induced with caterpillar regurgitant than seedlings not exposed to GLV. The use of pure synthetic chemicals revealed that (Z)-3 hexenal, (Z)-3-hexen-1-ol, and (Z)-3-hexenyl acetate have nearly identical priming activity. Caterpillar-induced nocturnal volatiles, which are enriched in GLV, also exhibited a strong priming effect, inducing production of larger amounts of JA and release of greater quantities of volatile organic compounds after caterpillar regurgitant application. In contrast, GLV priming did not affect JA production induced by mechanical wounding alone. Thus, GLV specifically prime neighboring plants against impending herbivory by enhancing inducible chemical defense responses triggered during attack and may play a key role in plant-plant signaling and plant-insect interactions. PMID- 14749517 TI - Microtransplantation of functional receptors and channels from the Alzheimer's brain to frog oocytes. AB - About a decade ago, cell membranes from the electric organ of Torpedo and from the rat brain were transplanted to frog oocytes, which thus acquired functional Torpedo and rat neurotransmitter receptors. Nevertheless, the great potential that this method has for studying human diseases has remained virtually untapped. Here, we show that cell membranes from the postmortem brains of humans that suffered Alzheimer's disease can be microtransplanted to the plasma membrane of Xenopus oocytes. We show also that these postmortem membranes carry neurotransmitter receptors and voltage-operated channels that are still functional, even after they have been kept frozen for many years. This method provides a new and powerful approach to study directly the functional characteristics and structure of receptors, channels, and other membrane proteins of the Alzheimer's brain. This knowledge may help in understanding the basis of Alzheimer's disease and also help in developing new treatments. PMID- 14749518 TI - Adverse drug reactions to anthelmintics. AB - BACKGROUND: Anthelmintics are commonly used in tropical areas, but are also prescribed in Western countries with other climates. However, pharmacoepidemiologic studies of these drugs are rare. OBJECTIVE: To investigate adverse drug reactions (ADRs) due to anthelmintics. METHODS: All spontaneous reports of ADRs associated with albendazole, diethylcarbamazine, flubendazole, ivermectin, mebendazole, niclosamide, praziquantel, pyrantel pamoate, and thiabendazole were identified in the French Pharmacovigilance Database from January 1, 1985, to August 31, 1999. For each case, the following data were recorded: age, gender, weight, ADRs, drug, dosage, and indication. RESULTS: A total of 243 cases were found corresponding to 291 ADRs. Serious ADRs (hematologic or hepatic injury) to albendazole most often occurred when the drug was used for the treatment of echinococcosis or cysticercosis, thus requiring both high dosage and long duration of therapy. Our data show that the profile and seriousness of anthelmintic-induced ADRs vary according to their use. Furthermore, the low number of spontaneous reporting of ADRs suggests a high rate of underreporting for these drugs, which are often considered in France as orphan drugs. CONCLUSIONS: The improvement of reporting of serious or unrecognized (unlabeled) anthelmintic-induced ADRs will increase our knowledge of the benefit/risk ratio associated with these agents and optimize their use. PMID- 14749519 TI - When spacing out brings clarity. Mental lapses serve as diagnostic clue for type of dementia. AB - NEW YORK CITY--Everyone has slow-witted moments: entering a room only to forget why, putting ice cream into the fridge, or blanking out on how to use the microwave. Some clinicians think that periodic mental lapses might help diagnose dementia with Lewy bodies (DLB), a scourge that kills brain cells (see DLB Case Study), although not all experts agree. Now, scientists have found that some cognitive blunders crop up more frequently in patients with DLB than in those with Alzheimer's disease (AD) or in healthy elderly individuals, according to work presented here on 15 January 2004 at an American Medical Association briefing for media. PMID- 14749520 TI - Plaque attack. Mice without ApoE and related protein unexpectedly accumulate beta amyloid deposits. AB - A person can toss her wadded-up burger wrapper into either the gutter or a trash bin. New work establishes that ApoE, a protein connected to Alzheimer's disease (AD), makes a similar choice. Mouse studies have suggested that ApoE encourages formation of the beta-amyloid plaques that gum up patients' brains. But scientists now suggest that ApoE helps clear beta amyloid from the brain before it forms plaques. The results warn that targeting ApoE as an AD treatment will require balancing its positive and negative influences. PMID- 14749521 TI - Hampering a heartbreaker. Antibiotic might stem injury from heart attack. AB - A TV ad urges people who think they're having a heart attack to pop an aspirin before rushing to the emergency room. They might be even better off taking antibiotics, according to a new study. The work shows that an antibiotic stems a previously untreatable form of heart damage not by killing bugs but by suppressing cellular enzymes. PMID- 14749522 TI - Frailty--the search for underlying causes. AB - Research into the biological basis of frailty has been difficult to accomplish because of a lack of standardized definitions, disease and disability confounders, and complex multifactorial etiology. Multiple physiological systems are likely to be involved, including the skeletal muscle, endocrine, and immune/inflammation systems. Physiological characterization of frail older adults might provide etiologic clues. Translational research programs that connect mechanisms related to aging, such as oxidative damage and telomere shortening, to clinical aging-related syndromes will be necessary to further this critical area of geriatric research. PMID- 14749523 TI - Resveratrol derivatives potently induce apoptosis in human promyelocytic leukemia cells. AB - Resveratrol has been shown to possess antioxidant and anticancer activities, but little is known on the effect of resveratrol derivatives. Recently we have isolated resveratrol and its dimers and trimers from peony (Paeonia lactiflora) seeds, and reported their strong antioxidant and cytotoxic activity. In the present study, we have evaluated cellular effects of resveratrol derivatives; viniferin, gnetin H, and suffruticosol B on the proliferation and apoptosis in HL 60 cells in vitro. All resveratrol and its derivatives reduced viability of HL-60 cells in a dose-dependent manner with their IC(50) values of 20-90 microM. Ascending orders of IC(50) values were suffruticosol B, gnetin H, viniferin and resveratrol respectively. HL-60 cells treated with the four stilbenes exhibited the distinct morphological changes characteristics of cell apoptosis such as chromatin condensation, apoptotic bodies, and DNA fragmentations. A time dependent histogram of the cellular DNA analyzed by flow cytometry revealed a rapid increase in subdiploid cells and a concomitant decrease in diploid cells exposed to 100 microM resveratrol for 0-24 h. Cells treated with 25 microM of resveratrol, viniferin, gnetin H, and suffruticosol B for 24 h resulted in increment of sub-G1 population by 51, 5, 11 and 59%, respectively. Treatment of cells with 0-20 microM resveratrol for 5 h produced a concentration-dependent decrease in cytochrome P450 (CYP) 1B1 mRNA levels. Suffruticosol B also suppressed CYP1B1 gene expression. These results demonstrated that resveratrol oligomers also strongly suppressed HL-60 cell proliferation, and induced DNA damage. In addition, CYP1B1 gene supression may suggest an involvement in the resveratrol-induced apoptosis in HL-60 cells. PMID- 14749524 TI - Biochemical properties of full-length hepatitis C virus RNA-dependent RNA polymerase expressed in insect cells. AB - The hepatitis C virus (HCV) RNA-dependent RNA polymerase, NS5B protein, is the key viral enzyme responsible for replication of the HCV viral RNA genome. Although several full-length and truncated forms of the HCV NS5B proteins have been expressed previously in insect cells, contamination of host terminal transferase (TNTase) has hampered analysis of the RNA synthesis initiation mechanism using natural HCV RNA templates. We have expressed the HCV NS5B protein in insect cells using a recombinant baculovirus and purified it to near homogeneity without contaminated TNTase. The highly purified recombinant HCV NS5B was capable of copying 9.6-kb full-length HCV RNA template, and mini-HCV RNA carrying both 5'- and 3'-untranslated regions (UTRs) of the HCV genome. In the absence of a primer, and other cellular and viral factors, the NS5B could elongate over HCV RNA templates, but the synthesized products were primarily in the double stranded form, indicating that no cyclic replication occurred with NS5B alone. RNA synthesis using RNA templates representing the 3'-end region of HCV minus-strand RNA and the X-RNA at the 3'-end of HCV RNA genome was also initiated de novo. No formation of dimer-size self-primed RNA products resulting from extension of the 3'-end hydroxyl group was observed. Despite the internal de novo initiation from the X-RNA, the NS5B could not initiate RNA synthesis from the internal region of oligouridylic acid (U)(20), suggesting that HCV RNA polymerase initiates RNA synthesis from the selected region in the 3'-UTR of HCV genome. PMID- 14749525 TI - Nonspecific association of 2',3'-cyclic nucleotide 3'-phosphodiesterase with the rat forebrain postsynaptic density fraction. AB - The 2',3'-cyclic nucleotide 3'-phosphodiesterase (CNP), a protein of unknown function in vivo, is abundantly expressed in myelinating glia in two isoforms, CNP1 and CNP2. In this study, immunoblot analysis showed that CNP1 is the major isoform in adult forebrain, and that both isoforms are included in the postsynaptic density (PSD) fraction and tyrosine-phosphorylated at the basal level. However, subcellular distribution and detergent extraction data showed that CNP is nonspecifically associated with the PSD fraction. Immunocytochemistry revealed that CNP is detected, in a weak but punctate pattern, in dissociated rat hippocampal neurons of 3 days to 2 weeks in vitro. The CNP-positive punctae were distributed throughout soma and dendrites, and distinct from PSD95-positive ones. Immunoblot analysis indicated that CNP is also expressed in neuronal stem cell lines, HiB5 and F11. Interestingly, in addition to the known two isoforms, a new CNP isoform of MW 45 kDa was expressed in these cell lines and was the major type of isoform in F11 cells. Taken together, our data suggest that CNP is expressed in the early stage of in vitro development and nonspecifically included in the adult rat PSD fraction. PMID- 14749526 TI - Adenoviral mediated hepatocyte growth factor gene attenuates hyperglycemia and beta cell destruction in overt diabetic mice. AB - Hepatocyte growth factor (HGF) is a potent mitogen and promoter of proliferation of insulin producing beta cells of pancreatic islets. To study the role of HGF, an adenoviral vector carrying the human HGF (Ad.hHGF) gene was transfected into the streptozotocin-induced diabetic mice and evaluated the effect on the blood glucose metabolism and the insulin-secreting beta cells of pancreatic islets. Ad.hHGF gene transfection resulted in amelioration of hyperglycemia and prolongation of survival period in the diabetic mice. Concomitantly adenoviral- mediated hHGF gene therapy slightly increased serum insulin concentration and the expression of insulin in the pancreatic islet. Although the proliferation of beta cell mass was not noticeable, the beneficial effect of HGF is significant to an almost deteriorated pancreatic islets. Taken together, these data suggest that the Ad.hHGF gene therapy into diabetic mice may prevent the further destruction and present as a beneficial remedy for type 1 diabetic patients. PMID- 14749527 TI - High levels of soluble herpes virus entry mediator in sera of patients with allergic and autoimmune diseases. AB - Herpes virus entry mediator (HVEM) is a newly discovered member of the tumor necrosis factor receptor (TNFR) superfamily that has a role in herpes simplex virus entry, in T cell activation and in tumor immunity. We generated mAb against HVEM and detected soluble HVEM (SHVEM) in the sera of patients with various autoimmune diseases. HVEM was constitutively expressed on CD4(+) and CD8(+) T cells, CD19(+) B cells, CD14(+) monocytes, neutrophils and dendritic cells. In three-way MLR, mAb 122 and 139 were agonists and mAb 108 had blocking activity. An ELISA was developed to detect sHVEM in patient sera. sHVEM levels were elevated in sera of patients with allergic asthma, atopic dermatitis and rheumatoid arthritis. The mAbs discussed here may be useful for studies of the role of HVEM in immune responses. Detection of soluble HVEM might have diagnostic and prognostic value in certain immunological disorders. PMID- 14749528 TI - Constitutive expression of 4-1BB on T cells enhances CD4+ T cell responses. AB - 4-1BB, a transmembrane molecule, member of the tumor necrosis factor receptor superfamily, is an important costimulatory molecule in the immune response, plays a key role in the clonal expansion and survival of CD8(+) T cells. In this study, we investigated 4-1BB regulation of CD4(+) T cell responses using 4-1BB transgenic (TG) mice that constitutively expressed 4-1BB on mature T cells. We first showed that CD4(+) T cells of 4-1BB TG mice had more sustained proliferative capacity in response to TCR/4-1BB stimulation in vitro compared to WT mice. Secondly, 4-1BB TG mice exhibited a more elevated contact hypersensitivity (CHS) response mediated by CD4+ Th1 cells due to more vigorous expansion of and apoptotic inhibition of CD4(+) T cells. Finally, CD4(+) T cells of 4-1BB TG mice had a heightened capacity for T cell priming. Overall, our results demonstrate the involvement of 4-1BB in CD4(+) Th1 cell responses by regulating the clonal expansion and survival of CD4(+) T cells as seen in CD8(+) T cells. PMID- 14749529 TI - Atrophy of brown adipocytes in the adult mouse causes transformation into white adipocyte-like cells. AB - Adipose tissue is an important endocrine regulator of glucose metabolism and energy homeostasis. Researches have focused on this tissue not only as a target for pharmacotherapy of obesity and insulin resistance but also as an endocrine tissue with leptin secretion and high insulin sensitivity. Brown adipose tissue (BAT) additionally plays a unique role in thermoregulation through the mitochondrial uncoupling protein 1 (UCP1), which uncouples oxidative phosphorylation. As a genetic tissue ablation model of BAT, we made transgenic mice expressing herpes simplex virus thymidine kinase (HSV-TK) driven by the brown adipocyte- specific UCP1 minimal regulatory element. The HSV-TK transgene was expressed specifically in BAT and more than 35% increase of apoptosis was induced by ganciclovir (GCV) treatment. Nevertheless, the expression level was not high enough to induce BAT ablation in GCV-treated adult mice. Importantly, however, we found that brown adipocytes in the periphery of interscapular BAT were transformed into white adipocyte-like unilocular cells. These cells express white adipocyte-specific leptin protein but are different in the ultrastructure of mitochondria from classical white adipocytes. Our data indicates that atrophy of BAT causes transformation into white adipocyte-like cells in the adult mouse and also suggests that further molecular understanding of adipocyte plasticity using our transgenic mouse model might be beneficial for the development of anti obesity/anti-diabetic therapies. PMID- 14749530 TI - The binding of ethidium bromide with DNA: interaction with single- and double stranded structures. AB - The pH-induced helix-coil transition of DNA and its complexes with EtBr is carried out at acidic pH in a wide interval of change of concentration ratio of EtBr/DNA. The binding isotherms of EtBr on double and single-stranded DNA at pH = 7.0 and pH = 3.0 (t = 25(o)C) are obtained by absorption and fluorimetric methods. Binding constants (K) and number of bases (n), corresponding to one binding site were determined. Non fluorescent "strong" complex with ds-DNA at pH = 7.0 and t = 25(o)C as well as "strong" and "weak" complexes with ss-DNA at pH = 3.0 and t = 25(o)C are revealed. PMID- 14749531 TI - Melting of complexes of DNA-cis-DDP in acidic environment. AB - The peculiarities of helix-coil transition of DNA at complex-formation with cis diamminedichlorplatinum (II) (cis-DDP) have been investigated in acidic environment by UV absorbance technique. It is shown that observed features of behavior T (pH) and T(m) (pH) DNA at pH 2.8-3.0 are possible to explain by formation in DNA of pseudo-ring structures at covalent linking of cis-DDP with DNA. PMID- 14749532 TI - Dysferlin in a hyperCKaemic patient with caveolin 3 mutation and in C2C12 cells after p38 MAP kinase inhibition. AB - Dysferlin is a plasma membrane protein of skeletal muscle whose deficiency causes Miyoshi myopathy, limb girdle muscular dystrophy 2B and distal anterior compartment myopathy. Recent studies have reported that dysferlin is implicated in membrane repair mechanism and coimmunoprecipitates with caveolin 3 in human skeletal muscle. Caveolin 3 is a principal structural protein of caveolae membrane domains in striated muscle cells and cardiac myocytes. Mutations of caveolin 3 gene (CAV3) cause different diseases and where caveolin 3 expression is defective, dysferlin localization is abnormal. We describe the alteration of dysferlin expression and localization in skeletal muscle from a patient with raised serum creatine kinase (hyperCKaemia), whose reduction of caveolin 3 is caused by a CAV3 P28L mutation. Moreover, we performed a study on dysferlin interaction with caveolin 3 in C2C12 cells. We show the association of dysferlin to cellular membrane of C2C12 myotubes and the low affinity link between dysferlin and caveolin 3 by immunoprecipitation techniques. We also reproduced caveolinopathy conditions in C2C12 cells by a selective p38 MAP kinase inhibition with SB203580, which blocks the expression of caveolin 3. In this model, myoblasts do not fuse into myotubes and we found that dysferlin expression is reduced. These results underline the importance of dysferlin-caveolin 3 relationship for skeletal muscle integrity and propose a cellular model to clarify the dysferlin alteration mechanisms in caveolinopathies. PMID- 14749533 TI - Angiotensin converting enzyme I/D, angiotensinogen T174M-M235T and angiotensin II type 1 receptor A1166C gene polymorphisms in Turkish hypertensive patients. AB - Essential hypertension is a multifactorial disease in which genetic and enviromental factors play an important role. These factors differ in each population. As there are no existing data for the Turkish population, we investigated four Renin Angiotensin System (RAS) gene polymorphisms, the angiotensin converting enzyme (ACE), angiotensinogen (AGN) M235T/T174M and angiotensin II type 1 receptor A1166C polymorphism in 109 hypertensive and 86 normotensive Turkish subjects. Polymerase Chain Reaction (PCR) and Restriction Fragment Length Polymorphism (RFLP), and agarose gel electrophoresis tecniques were used to determine these polymorphism. The frequencies of person that carry ACE D allel (DD+ID) was significantly higher in hypertensive group (99.1%) than controls (80%) (P 0.000). M235T TT genotype was also found significantly higher in hypertensives than control group (20% vs 2.7%; P 0.001). The frequency of AGN 174M allele was higher in the hypertensive group than control subjects (8.76% vs 4.81%). Frequency of ATR1 C allele (AC+CC genotypes) was found higher hypertensives than controls (39.4% vs 25.9%; P = 0.054). Our results suggest that an interaction exists between the RAS genes and hypertension in Turkish population. PMID- 14749534 TI - Vitamin D receptor gene polymorphisms in breast cancer. AB - Breast cancer is the leading cause of cancer death among women around the world and its incidence is annually increasing. The vitamin D receptor (VDR) gene is a member of the nuclear receptor superfamily, which is expressed in breast tissue and known to modulate the rate of cell proliferation. Association between the VDR gene polymorphisms and cancer development has been suggested by several studies. However, the relationship between VDR polymorphisms and breast cancer is controversial and has not been confirmed by all studies. The purpose of this study was to investigate the genotype frequencies and association of the VDR Bsm I and Taq I polymorphisms with breast cancer in Turkish patients. In this study, 78 patients with breast cancer and 27 healthy individuals were enrolled. The prevalence of the VDR Taq I and Bsm I alleles and the genotype frequencies in patients with breast cancer was similar to that in the normal population. Our data indicate that no significant differences exist between the patients and control subjects. PMID- 14749535 TI - Protein kinase CK2 phosphorylates and interacts with deoxyhypusine synthase in HeLa cells. AB - Deoxyhypusine is a modified lysine and formed posttranslationally to be the eukaryotic initiation factor eIF5A by deoxyhypusine synthase, employing spermidine as butylamine donor. Subsequent hydroxylation of this deoxyhypusine containing intermediate completes the maturation of eIF5A. The previous report showed that deoxyhypusine synthase was phosphorylated by PKC in vivo and the association of deoxyhypusine synthase with PKC in CHO cells was PMA-, and Ca(2+)/phospholipid-dependent. We have extended study on the phosphorylation of deoxyhypusine synthase by protein kinase CK2 in order to define its role on the regulation of eIF5A in the cell. The results showed that deoxyhypusine synthase was phosphorylated by CK2 in vivo as well as in vitro. Endogenous CK2 in HeLa cells and the cell lysate was able to phosphorylate deoxyhypusine synthase and this modification is enhanced or decreased by the addition of CK2 effectors such as polylysine, heparin, and poly(Glu, Tyr) 4:1. Phosphoamino acid analysis of this enzyme revealed that deoxyhypusine synthase is mainly phosphorylated on threonine residue and less intensely on serine. These results suggest that phosphorylation of deoxyhypusine synthase is CK2-dependent cellular event as well as PKC-mediated effect. However, there were no observable changes in enzyme activity between the phosphorylated and unphosphorylated forms of deoxyhypusine synthase. Taken together, besides its established function in hypusine modification involving eIF5A substrate, deoxyhypusine synthase and its phosphorylation modification may have other independent cellular functions because of versatile roles of deoxyhypusine synthase. PMID- 14749536 TI - Role of crosslinked protein in lung injury following total body irradiation and bone marrow transplantation. AB - The aberrant protein crosslinks formation during lung injury as results total body irradiation (TBI) and bone marrow transplantation (BMT) therapy has been examined as apossible contributory factor in organ or tissue pathogenesis. Female C3HeB/ FeJ mice were used for an experimental animal. Carbon monoxide uptake (V(CO)) was measured at 1, 2, 3, 4 and 5 months after TBI at respective doses of 12, 14, 16 and 18 Gy 16 h prior to syngeneic BMT. Also as a measure of aberrant protein crosslinking in the inured tissues, transglutaminase (TGase)-activities and crosslinked protein were examined along with thrombin, a protease known to activate TGases. Reductions of VCO were detected following TBI and BMT. Activities of thrombin and TGase 1, and crosslinked protein in bronchoalveolar lavage (BAL) fluid of the mice 1 wk after TBI at 12 Gy and BMT were identified and found to be elevated in the treated animals. These findings suggest that elevated levels of crosslinked proteins and TGase I in the bronchoalveolar larvage during the lung injury could have enhanced the organ pathogenesis following TBI and BMT. PMID- 14749537 TI - Targeting efficiency of a-1,3-galactosyl transferase gene in pig fetal fibroblast cells. AB - Animal cloning technology with somatic cells provides an alternative tool to conventional methods for producing transgenic animals. Gene targeting in animals is made feasible using somatic cells with homologous recombination procedure that is a major technique in embryonic stem cells for knocking-out genes. Homologous recombination events in somatic cells are relatively inefficient as compared to those in ES cells, suggesting the need for establishment of efficient gene targeting system in somatic cells. To investigate the efficiency of positive and negative selection for gene targeting in pig fetal fibroblast cells, pig alpha 1,3-galactosyl transferase (13-GT) gene was used for gene targeting. The neomycin phosphotransferase (Neo(r)) and herpes simplex virus-thymidine kinase (HSV-tk) genes were used as positive and negative selection markers in this experiment. Following transfection with targeting DNA construct, the pig fetal fibroblast cells were selected against resistance of G418 and gancyclovir. In DMEM medium containing 5 to 10% serum, Pig fetal fibroblast cells failed to proliferate during drug selection. Increasing serum concentration to 15% of medium yielded less senescent colonies of pig fetal fibroblast cells following drug selection that allowed enough cell colonies to screen genomic DNA. The frequency of gene targeting in pig fetal fibroblast cells with double drug selection was more than 10-fold efficient compared to that with G418 single selection. Double selection method with Neo' and HSV-tk genes could be useful for gene targeting in somatic cells for production of cloned animals carrying targeted endogenous genes. PMID- 14749539 TI - An improved method for constructing a full-length enriched cDNA library using small amounts of total RNA as a starting material. AB - We have developed an improved method for constructing a full-length cDNA library using small quantity of material by modifying the original oligo-capping method. In our devised method, total RNAs are used in sequential oligo-capping steps directly without preliminary mRNA purification. Using this method, we constructed full- length cDNA libraries from 100 mg of total RNA. These libraries contained 8x10(5) to 8x10(6) independent clones with average insert sizes of 2.0 kb. Moreover, the number of full-length cDNAs containing the translation initiation codon ATG in the constructed libraries was estimated to 60-70%. In addition, 54% of the known cDNAs had a longer 5' end than the corresponding genes in the public database. Our results show that the method can be effectively used to construct full-length enriched cDNA libraries, especially, if starting material is limited. PMID- 14749538 TI - Differential inhibition of endothelial cell proliferation and migration by urokinase subdomains: amino-terminal fragment and kringle domain. AB - The serine protease urokinase-type plasminogen activator (uPA) is implicated in pericellular proteolysis in a variety of physiological and pathological processes including angiogenesis and tumor metastasis. The kringle domain of uPA (UK1) has proven to be an anti-angiogenic molecule with unknown mechanism and amino terminal fragment of uPA (u-ATF) with additional growth factor-like domain can be used for blocking interaction of uPA and uPA receptor. Here, we compared anti angiogenic activities of these two molecules in vitro and in vivo. The recombinant u-ATF from E. coli and refolded in vitro was found to bind to uPAR with high affinity, whereas E. coli-derived UK1 showed no binding by Biacore analysis. In contrast to UK1 having potent inhibitory effect, u-ATF exhibited low inhibitory effect on bovine capillary endothelial cell growth (ED(50)>320 nM). Furthermore, u-ATF inhibition of VEGF-induced migration of human umbilical vein endothelial cell was far less sensitive (IC(50) = 600 nM) than those observed with UK1, and angiogenesis inhibition was marginal in chorioallantoic membrane. These results suggest that kringle domain alone is sufficient for potent anti- angiogenic activity and additional growth factor-like domain diverts this molecule in undergoing different mechanism such as inhibition of uPA/uPAR interaction rather than undergoing distinct anti- angiogenic mechanism driven by kringle domain. PMID- 14749540 TI - Lack of p38 MAP kinase activation in TRAIL-resistant cells is not related to the resistance to TRAIL-mediated cell death. AB - Activation of MAP kinases is involved in various cellular processes, including immunoregulation, inflammation, cell growth, cell differentiation, and cell death. To investigate the role of p38 MAP kinase activation in the signaling pathway of TRAIL-mediated apoptosis, we compared TRAIL-mediated MAP kinase activation in TRAIL-susceptible human colon cancer cell line DLD1 and TRAIL resistant DLD1/TRAIL-R cells. TRAIL-mediated activation of ERK occurred in both cell lines. In contrast, both DLD1 and DLD1/TRAIL-R cells showed no obvious JNK activation after treatment with TRAIL. Interestingly, TRAIL-mediated activation of p38 MAP kinases was observed in DLD1 cells but not in DLD1/TRAIL-R cells. However, activation of p38 MAP kinases was observed in both DLD1 and DLD1/TRAIL-R cells after treatment with anisomycin. Furthermore, inhibiting activated p38 MAP kinases with known inhibitors or with an adenovector expressing dominant negative p38alpha did not block TRAIL-mediated cell death in DLD1 cells. Moreover, activation of p38 MAP kinases by adenovectors expressing constitutive MKK3 or MKK6 (Ad/MKK3bE or Ad/MKK6bE) did not induce cell death in either DLD1 or DLD1/TRAIL-R cell lines. Our results suggest that activation of p38 MAP kinases does not play a major role in TRAIL-mediated apoptosis in DLD1 cells and that lack of TRAIL-mediated p38 MAP kinase activation may not be the mechanism of TRAIL-resistance in DLD1/TRAIL-R cells. PMID- 14749542 TI - Pharmacokinetics of omeprazole in healthy adults and in children with gastroesophageal reflux disease. AB - Studies of the pharmacokinetics of omeprazole in children with gastroesophageal reflux disease (GERD) remain scarce despite the vast number of reports on its efficacy. The objectives of this study were to assess the pharmacokinetics of omeprazole in healthy adults and in children with GERD. Omeprazole (Losec, delayed-release capsules) was administered orally to 18 healthy adults (mean age 36.8 years) and 12 children with GERD (mean age 6.1 years). Blood samples were collected over 5 hours, and plasma concentrations were assessed using liquid chromatography. Population pharmacokinetic parameters were calculated using NONMEM. A 1-compartment model with zero-order absorption and a lag time was used. The population approach was well suited to the limited number of samples available, and residual variability was low. Oral clearance (CL/F) and apparent volume of distribution (V(ss)/F) in healthy adults (Mean +/- SD: 0.62 +/- 0.27 L/h/kg and 0.76 +/- 0.26 L/kg, respectively) were not significantly different than those in children with GERD (0.51 +/- 0.34 L/h/kg and 0.66 +/- 0.25 L/kg, respectively). Healthy adults displayed a statistically significantly longer delay in drug absorption (Lag time: 0.62 +/- 0.15 hours) as compared with that observed in children with GERD (0.12 +/- 0.03 hours, P < 0.05). On the basis of these findings, omeprazole dosings on a milligram-per-kilogram basis are recommended with no further adjustments for the treatment of GERD in children. PMID- 14749543 TI - Dosage adjustment of ribavirin based on renal function in Japanese patients with chronic hepatitis C. AB - The daily dose of ribavirin is currently determined based on body weight. In the present study, the authors examined factors influencing total plasma clearance (CL(total)) and the toxic level on red blood cells of ribavirin in such body weight-based dosage adjustment in Japanese chronic hepatitis C patients (13 male and 6 female). Patients received ribavirin (600 or 800 mg/d) orally, depending on their body weights, together with interferon alpha-2b (6 million units) intramuscularly. A steady-state trough plasma concentration (C(pss)) was achieved approximately 4 weeks after the initiation of treatment, but the value was scattered among patients in a range from 1100 to 4200 ng/mL. The high C(pss) of ribavirin of approximately 4000 ng/mL decreased hemoglobin concentrations to less than 8.5 g/dL. The individual CL(total), estimated by dividing dose normalized by body weight by C(pss), of ribavirin correlated significantly with the patient's creatinine clearance. In contrast, no relationship was observed with other parameters such as age, body weight, serum creatinine concentration, alanine aminotransferase (ALT) concentration, or aspartate aminotransferase (AST) concentration, though ALT and AST concentrations decreased with ribavirin treatment in most patients. These results indicate that CL(total) of ribavirin is dependent on renal function (creatinine clearance), and hemolysis is induced by high ribavirin concentrations in plasma. Dosage adjustment of ribavirin based on renal function and body weight would provide effective and safer treatment without causing hemolysis. PMID- 14749544 TI - Prednisolone: limited sampling strategies for estimating pharmacokinetic parameters. AB - To develop limited-sampling strategy (LSS) models for estimating prednisolone's area under plasma concentration versus time curve (AUC(0-infinity)), its maximum concentration in plasma (C(max)), and total clearance (CL/F). Healthy subjects (n = 24), enrolled in a bioequivalence study, received 20 mg PO of the prodrug prednisone as reference and test tablets, and plasma prednisolone concentrations (n = 576) were measured by a validated HPLC assay. A linear regression analysis of AUC(0-infinity), C(max), CL/F, and log(CL/F) against the plasma prednisolone concentrations for the reference formulation was carried out to develop LSS models to estimate these parameters. The LSS models were validated on the test formulation data sets and on simulated sets generated by the software ADAPT II. LSS models based on a single [1.5 hours for C(max) and 7 hours for AUC(0 infinity), CL/F, and log(CL/F)] plasma sample, accurately estimated (R2 = 0.84 0.97, mean bias < 1%; mean precision < 10%) these pharmacokinetic parameters. Validation tests indicated that the most informative single-point LSS models developed for the reference formulation provide precise estimates (R(2) > 0.83; mean bias < 3%; mean precision < 10%) of the corresponding pharmacokinetic parameters for the test formulation. LSS models based on the two most informative sampling points (1.5 and 7 hours) were required for accurate estimates (R(2) > 0.87; mean bias < 6%; mean precision < 8%) of prednisolone's C(max), AUC(0 infinity), CL/F, and log(CL/F) for the simulated data sets. Finally, bioequivalence assessment of the prednisone formulations, based on LSS-derived AUC(0-infinity) and C(max) values provided results identical to those obtained using the original values for these parameters. One- and 2-point LSS models provided accurate estimates of prednisolone's C(max), AUC(0-infinity), and CL/F, following single oral doses of prednisone, and allowed correct assessment of bioequivalence between two prednisone formulations. PMID- 14749546 TI - The effect of parenteral nutrition fluids on the binding of therapeutic drugs to human serum in vitro. AB - The competitive binding of seven therapeutic drugs (carbamazepine, phenytoin, phenobarbital, procainamide, quinidine, theophylline, and valproic acid) to human serum and to five commonly used parenteral nutrition fluids in vitro was studied using equilibrium dialysis. For five of the drugs, all parenteral nutrition fluids bound less drug than human serum-phenobarbital (up to 14% less), phenytoin (up to 46% less), procainamide (up to 43% less), quinidine (up to 25% less), and valproic acid (up to 77% less)-suggesting that the presence of these fluids might increase the free fraction of these drugs in vivo. For carbamazepine, the fluids bound up to 82% more drug, suggesting that the presence of these fluids might decrease the free fraction of this drug in vivo. For theophylline, the fluids produced a minimal (no more than 5%) effect on binding to serum. The administration of parenteral nutrition fluids may significantly alter the free (active) fraction of some therapeutic drugs. PMID- 14749545 TI - Population pharmacokinetic modeling of oral cyclosporin using NONMEM: comparison of absorption pharmacokinetic models and design of a Bayesian estimator. AB - There have been very few population pharmacokinetic (PopPK) studies and Bayesian forecasting methods dealing with cyclosporin (CsA) so far, probably because of the difficulty of modeling the particular absorption profiles of CsA. The present study was conducted in stable renal transplant patients treated with Neoral and employed the NONMEM program. Its goals were (1) to develop a population pharmacokinetic model for CsA based on an Erlang frequency distribution (which describes asymmetric S-shaped absorption profiles) combined with a 2-compartment model; (2) to compare this model with models combining a time-lag parameter and either a zero-order or first-order rate constant and with a model based on a Weibull distribution; and (3) to develop a PK Bayesian estimator for full AUC estimation based on that "Erlang model." The PopPK model was developed in an index set of 70 patients, and then individual PK parameters and AUC were estimated in 10 other patients using Bayesian estimation. The "Erlang" model best described the data, with mean absorption time (MAT), apparent clearance (CL/F), and apparent volume of the central compartment (Vc/F) of 0.78 hours, 26.3 L/h, and 76 L, respectively (interindividual variability CV = 33, 30, and 48%). Bayesian estimation allowed accurate prediction of systemic exposure using only 3 samples collected at 0, 1, and 3 hours. Regression analysis found no significant difference between the predicted and observed concentrations (10 per patient), and AUC(0-12) were estimated with a nonsignificant bias (0.6 to 8.7%) and good precision (RMSE = 5.3%). In conclusion, the Erlang distribution best described CsA absorption profiles, and a Bayesian estimator developed using this model and a mixed-effect PK modeling program provided accurate estimates of CsA systemic exposure using only 3 blood samples. PMID- 14749547 TI - Lopinavir protein binding in vivo through the 12-hour dosing interval. AB - Most protease inhibitors available for the treatment of human immunodeficiency virus (HIV) infection are highly bound to plasma proteins, mainly alpha-1 acid glycoprotein. Therapeutic drug monitoring (TDM) of total protease inhibitor (PI) concentrations has been increasing in the past few years; however, the pharmacological activity of the PIs is dependent on unbound drug entering cells harboring HIV. There is little information available on unbound drug concentrations of these drugs in vivo. The aim of the study was to measure unbound plasma concentrations of lopinavir (LPV) and to relate them to the total plasma concentrations to establish the unbound percentage in vivo during a full dosage interval. A pharmacokinetic study was performed in HIV-infected subjects (n = 23; median CD4 cell count = 290 x 10(6) cells x L(-1); viral load < 50 copies x mL(-1)) treated with a LPV/ritonavir (RTV)-containing regimen. Ultrafiltration was used to separate unbound LPV in all plasma samples (n = 115). Equilibrium dialysis was also used to compare with ultrafiltration measurements in 10/23 patients at baseline and 2 hours after drug intake. Total and unbound LPV concentrations were measured by a fully validated method using high performance liquid chromatography-mass spectometry (HPLC-MS/MS). Based on a comparison of AUC(unbound)AUC(total), the mean (+/- SD) unbound percentage of LPV from all the samples studied (n = 115) was 0.92% (+/- 0.22) when measured with ultrafiltration and 1.32% (+/- 0.44) when equilibrium dialysis was used (n = 20), showing a higher drug recovery (P = 0.048). The unbound percentage of LPV was found to be significantly higher after 2 h than at baseline (P < 0.05 with both methods), suggesting a concentration-dependent binding of LPV that has not been observed in vitro. However, the clinical significance of such phenomena is still unclear. PMID- 14749548 TI - The binding of selected therapeutic drugs to human serum alpha-1 acid glycoprotein and to human serum albumin in vitro. AB - The binding of acetaminophen, lidocaine, phenobarbital, phenytoin, theophylline, and valproic acid to human serum alpha-1 acid glycoprotein (orosomucoid) and to human serum albumin separately in vitro was investigated using equilibrium dialysis of the unlabeled drugs. Each drug was studied at a therapeutic concentration. Alpha-1 acid glycoprotein was studied at one elevated and two physiological concentrations, whereas albumin was studied at one physiological and two low concentrations. The nonphysiological concentrations were consistent with those that might be seen in a variety of clinical conditions. Acetaminophen, phenobarbital, theophylline, and valproic acid showed negligible binding to alpha 1 acid glycoprotein. However, lidocaine and phenytoin demonstrated binding to this protein, and increases in the alpha-1 acid glycoprotein concentration produced decreases in the unbound (free) or "active" concentration of these two drugs. All drugs but acetaminophen bound to albumin, and decreases in the albumin concentration yielded increases in the unbound (free) or "active" concentration of the remaining 5 drugs. These findings are significant when lidocaine, phenytoin, phenobarbital, theophylline, or valproic acid are used in patients with clinical conditions that may affect the concentration of these two binding proteins. PMID- 14749549 TI - Neuronal MDR-1 gene expression and persistent low levels of anticonvulsants in a child with refractory epilepsy. AB - SUMMARY: It is estimated that 20-25% of epileptic patients fail to achieve good control with antiepileptic drug (AED) treatment; thus, refractory epilepsy (RE) has been described in patients who have adequate therapeutic levels of AEDs without control of seizures. Multidrug resistance genes have been reported to be highly expressed in brain of patients with RE. Persistent low plasma levels of AEDs and high brain expression of the multidrug resistance product P-glycoprotein (P-gp) have been previously communicated in a case report of RE secondary to tuberous sclerosis. Here, the authors report a case of an 8-year-old boy diagnosed with partial RE with focal seizures who was admitted to hospital for a severe episode of subintrant crisis. The patient received polytherapy with carbamazepine (CBZ), phenytoin (PHT), and valproic acid (VA); however, habitual doses of these AEDs failed to control the patient's symptoms. AED blood levels were monitored for 25 consecutive days and showed low values in 8/25 (33%) for CBZ, 10/25 (40%) for PHT, and 25/25 (100%) for VA of samples studied. Because the patient developed focal status epilepticus, surgical treatment by callosotomy was done, resulting in a significant improvement in epileptic symptoms. The immunostaining of brain specimens showed significantly increased expression of P gp not only in vascular endothelial cells and related astrocytes but also in neurons. Overexpression of P-gp in the brain does not explain the low blood levels of AEDs described in these cases. Different mechanisms such as drug-drug interactions and drug transporters can be involved in the results observed. The P gp overexpression and/or its pharmacologic induction should be considered as a potential mechanism responsible for drug resistance to epilepsy treatment and highly suspected in patients with persistent subtherapeutic AEDs plasma levels. PMID- 14749550 TI - A novel system for the determination of antidepressant concentrations in human breast milk. AB - The high incidence of psychiatric illness in the postpartum period and the increasing percentage of women who breastfeed has focused attention on the treatment of breastfeeding women with psychotropic medications and, additionally, the exposure of nursing infants to these medications. Consequently, there has been an increased effort to develop standardized methods for quantifying psychotropic medications in breast milk. This paper details a novel method for quantifying the concentrations of multiple selective serotonin reuptake inhibitors (SSRIs) and tricyclic antidepressants (TCAs) in breast milk. The method consists of a common liquid/liquid and solid-phase extraction followed by HPLC separation on a common column and UV detection. Assay system 1 measures fluoxetine, norfluoxetine, fluvoxamine, and paroxetine; assay 2 measures sertraline and desmethylsertraline; and assay 3 measures the TCAs including doxepin, nordoxepin, desipramine, imipramine, nortriptyline, and amitriptyline. The method is shown to be a highly accurate and precise technique for measuring 12 different antidepressants in human breast milk and to be free of the matrix effects often encountered in breast milk drug analyses. PMID- 14749551 TI - Bioequivalence of a new cyclosporine a formulation to Neoral. AB - New cyclosporine A (CsA) formulations must prove their bioequivalence to Neoral, the reference CsA formulation, to allow free prescription for the patients. The aim of this study was to compare the pharmacokinetics (PK) of a new CsA formulation (Zinograf-ME), produced by Strides-Arcolab, to Neoral and to demonstrate their interchangeability in stable renal transplant recipients. Twelve-hour PK studies were obtained from 18 (13 M/5 F) adult patients (mean age 44.7 +/- 12 years). They received their renal allografts from 13 cadaver and 5 living donors. Before enrollment, all patients were receiving a third generic CsA for a mean of 48 months. Nine patients were also under azathioprine and 9 under mycophenolate mofetil; 17 received prednisone. A single oral dose of either Zinograf or Neoral was administered. The first PK study was performed with one formulation, and 1 week later, a second PK was done with the other formulation. During the washout period, patients continued taking the third CsA formulation. The drug substitution was done milligram-for-milligram. The CsA whole-blood level was measured by TDx immunoassay. Mean +/- SD of area under the curve (AUC), maximum concentration (C(max)), and concentration at the second hour (C2) of Zinograf were not statistically different from those with Neoral (4019 +/- 1466 vs 3971 +/- 1325 ng x h/mL, 998 +/- 376 vs 1021 +/- 356 ng/mL, and 707 +/- 254 vs 734 +/- 229 ng/mL, respectively). In the same way, the Zinograf 90% confidence interval for either C(max) (-123, +77 ng/mL) or AUC (-214, +311 ng.mL/h) were within the Neoral bioequivalence interval for the same parameters (+/-204 ng/mL and +/-794 ng x mL/h, respectively). These data demonstrate that the ZinografME CsA formulation is bioequivalent to Neoral. PMID- 14749552 TI - Monitoring cyclosporine of pre-dose and post-dose samples using nonextraction homogeneous immunoassay. AB - A nonextraction homogeneous immunoassay (CEDIA Cyclosporine Plus Assay) has been developed for the measurement of cyclosporine in predose (trough) and post-dose (C2 to C8) whole-blood samples. The method includes a low-range assay that measures cyclosporine from 25 to 450 ng/mL in pre-dose samples and a high-range assay that detects cyclosporine from 450 to 2000 ng/mL in post-dose samples. The high-range assay allows a direct measurement of post-dose samples without a dilution step. Alternatively, post-dose samples can be correctly measured by the low-range assay following a twofold dilution. Using an NCCLS precision protocol, the assay exhibited less than 10% CV or error less than the functional sensitivity. Functional sensitivity of the low-range assay was demonstrated at 20 ng/mL cyclosporine. Cross-reactivity was measured in the presence of cyclosporine and was found to be 4.4%, 19.8%, 16.4%, 0.9%, 1.0%, and 1.6% for metabolites AM1, AM9, AM4n, AM19, AM4n9, and AM1c, respectively. When 53 samples were evaluated using an HPLC method, the three most significant cross-reactive metabolites, AM1, AM4n, and AM9, exhibited an average concentration profile of 123%, 19%, and 0.06% of the parent cyclosporine, respectively. The average total contribution to cyclosporine quantification from these metabolites was estimated at 7.2% based on the percentage cross-reactivity of each metabolite in the CEDIA assay and the concentration of each metabolite as determined by HPLC. The method comparison study revealed a linear regression correlation of CEDIA = 1.095 x HPLC + 6.6, r = 0.972, for the low-range assay, and CEDIA = 1.018 x HPLC - 36.4, r = 0.968, for the high-range assay. In conclusion, the CEDIA Cyclosporine Plus Assay is a precise and accurate method for quantification of cyclosporine in pre-dose and post-dose samples. PMID- 14749553 TI - Ganciclovir in solid organ transplant recipients: is there a role for clinical pharmacokinetic monitoring? AB - The authors use a previously published decision-making algorithm to address the role of clinical pharmacokinetic monitoring of ganciclovir, the drug of choice for prophylaxis and treatment of cytomegalovirus (CMV) in solid organ transplant recipients. Ganciclovir pharmacokinetics have been studied in solid organ transplant recipients with a wide range of peak and trough concentrations reported. Numerous assays are available to measure plasma concentrations of ganciclovir, but no clear correlation has been established between peak or trough concentrations and either efficacy or toxicity of the drug. For patients receiving treatment, the pharmacological response of ganciclovir is assessed initially by clinical response. Monitoring prophylactic therapy in asymptomatic patients poses a greater challenge. Although monitoring of antigenemia or polymerase chain reaction (PCR) deoxyribonucleic acid (DNA) is not yet part of routine clinical practice, studies have shown a role for these techniques in monitoring response to antiviral therapy. Studies of subpopulations of renal failure patients show a prolonged ganciclovir half-life that requires dosage adjustments. However, ganciclovir clearance is closely correlated with creatinine clearance, which is an appropriate approach to adjusting dosages. Studies in pediatric patients also demonstrate a close correlation between dose per kilogram and AUC, suggesting that monitoring of ganciclovir levels may not be necessary. Based on the evidence presented in this review, routine clinical pharmacokinetic monitoring of ganciclovir does not appear to be warranted in solid organ transplant recipients. PMID- 14749554 TI - Therapeutic drug monitoring of the antidepressant mirtazapine and its N demethylated metabolite in human serum. AB - Mirtazapine is a novel antidepressant that acts by enhancing serotonergic and noradrenergic neurotransmission. Because very little is known about serum concentrations in relation to clinical effects, the use of therapeutic drug monitoring is so far unclear. A rapid automated HPLC method with fluorescence detection was developed for routine quantification of mirtazapine and its demethylated metabolite N-desmethylmirtazapine in human serum. The precision of the method was suitable because the day-to-day (n = 7) coefficient of variation (CV) of mirtazapine was 9.8, 4.2, and 5.1% for concentrations of 10, 40, and 80 ng/mL, respectively, and the CV for N-desmethylmirtazapine were 11.6, 10.3, and 9.5% for 5, 20, and 40 ng/mL, respectively. The bias ranged between 0.7 and 4.2 ng/mL and between 0.9 and 2.0 ng/mL for mirtazapine and N-desmethylmirtazapine, respectively. Serum samples of 100 patients, aged between 18 and 93 years, were analyzed. There was wide interindividual variability of serum concentrations on each dose level, and the median (25th to 75th percentiles) of the mirtazapine and N-desmethylmirtazapine concentrations was 19.5 (11.0-28.7) and 9.0 (6.0-17.0) ng/mL, respectively. Women had higher dose-corrected concentrations (C/Ds, ng/mL/mg) of mirtazapine [median (25th-75th percentiles) 0.6 (0.4-0.9) vs 0.4 (0.3-0.6) and N-desmethylmirtazapine [0.4 (0.2-0.6) vs 0.2 (0.1-0.4)] than men. Patients over 60 years of age (mean age +/- SD was 72.2 +/- 7.1) had higher C/Ds of mirtazapine and N-desmethylmirtazapine [0.7 (0.4-1.2) vs 0.53 (0.4-0.8) and 0.5 (0.2-0.9) vs 0.3 (0.2-0.9), respectively] than younger patients (mean age +/- SD was 43.3. +/- 10.6). Patients with N-desmethylmirtazapine/mirtazapine ratios less than 0.4 had significantly more side effects (P < 0.05) than those having higher ratios. Comedications were assessed for drug-drug interaction, and significantly (P < 0.05) lower N-desmethylmirtazapine/mirtazapine ratios were found under concomitant medications of the antidepressant sertraline and the antipsychotic amisulpride. PMID- 14749555 TI - Interference of endogenous digoxin-like immunoreactive factors in serum digoxin measurement is minimized in a new turbidimetric digoxin immunoassay on ADVIA 1650 analyzer. AB - Endogenous digoxin-like immunoreactive factors (DLIF) cross-react with antidigoxin antibody and falsely elevate or lower measured serum digoxin concentrations, depending on the assay design. Recently, Bayer Diagnostics released a turbidimetric assay for digoxin on the ADVIA 1650 analyzer. We studied potential interference of DLIF with this new digoxin assay. We analyzed 40 serum specimens from patients who have pathologic conditions that may increase serum DLIF concentrations. These patients were never exposed to digoxin or other agents that may lead to a measurable digoxin concentration. We also analyzed five specimens from autopsy and five specimens from neonates. Apparent digoxin concentrations were measured using the new turbidimetric digoxin assay, the fluorescence polarization immunoassay (FPIA, Abbott Laboratories, Abbott Park, IL), and also the chemiluminescent immunoassay (CLIA, Bayer Diagnostics). We observed measurable apparent digoxin levels with the FPIA in 5 uremic patients (range 0.24-0.86 ng/mL), 6 patients with liver disease (range 0.21-0.72 ng/mL), in 3 patients in the third trimester of pregnancy (0.21-26 ng/mL), and in 3 neonates (range 0.21-0.46 ng/mL). Four out of 5 autopsy specimens showed measurable apparent digoxin concentrations (0.23-0.81 ng/mL). In contrast, only 1 specimen (a uremic patient) showed an apparent digoxin concentration of 0.26 ng/mL with the turbidimetric digoxin immunoassay (FPIA value 0.86 ng/mL, CLIA value 0.32 ng/mL). Because DLIF is absent in the protein-free ultrafiltrate, we also measured free digoxin concentrations in DLIF-positive patients to ensure that the apparent digoxin concentrations were caused by DLIF. We observed no apparent digoxin concentrations in the protein-free ultrafiltrate in any DLIF positive specimens. When serum specimens containing elevated concentrations of DLIF but no digoxin were supplemented with a known concentration of digoxin, we observed falsely elevated digoxin concentrations by the FPIA, as expected. In contrast, we observed a good agreement between the target and observed concentrations when the new turbidimetric assay was used. We conclude that DLIF has minimal effect on serum digoxin measurements by the new turbidimetric assay. PMID- 14749556 TI - Multicomponent screening for drugs of abuse: direct analysis of urine by LC-MS MS. AB - By allowing for direct injection of urine, liquid chromatography-tandem mass spectrometry (LC-MS-MS) with atmospheric-pressure chemical ionization has proven to be useful for analysis of drugs of abuse in human urine. The purpose of this study was to evaluate this technique by direct screening for 23 different substances (phenylethylamines, hypnotics, and N-benzylpiperazine) in urine samples. It was possible to achieve lower detection limits compared with commercial immunochemical methods. There was a linear response for all analytes with an intraday coefficient of variation of about 16%, and the gradient elution gave a variability in relative retention time of about 1%. Positive results were confirmed by reanalysis including sample preparation by solid-phase extraction. Among the 529 authentic urine samples analyzed, 35 samples were screened positive for phenylethylamines, and 20 for hypnotics. Of these, 23 (66%) samples were confirmed to be positive for phenylethylamines, and 11 (55%) for hypnotics. This study demonstrates that LC-MS-MS is a valuable complement to immunochemical screening analysis, especially for substances for which immunochemical methods are not yet available or when an increased sensitivity is needed. PMID- 14749557 TI - Analytic evaluation and application of a novel spectrophotometric serum lithium method to a rapid response laboratory. AB - The authors present an evaluation of the Lithium DST spectrophotometric method developed by Thermotrace (Victoria, Australia) on a Hitachi 917 analyzer. Accuracy was assessed by method comparison with an ion-selective electrode (ISE; Roche Integra 700) (n = 80). Linearity, within-run and between-run precision, and susceptibility to interference by hemolysis, icterus, lipemia, and sodium were assessed. The method was linear to 3.0 mM/L, and analyzer auto-dilution extended the reportable range to 7.2 mmol/L. Within-run coefficient of variation was 1.5% at 0.68 mmol/L and 0.7% at 2.06 mM/L. Between-run precision for the same lithium concentrations assayed daily for 20 days were 3.1% and 1.9%, respectively. Method agreement with ISE was excellent, with an intercept of 0.000 and slope of 1.000 by Passing Bablock regression analysis. Hemolysis, icterus, lipemia, and high and low sodium levels did not significantly interfere. The manufacturer's recommended calibration stability of 1 week was confirmed. The authors conclude that this method is reliable, accurate, and precise. The Thermotrace serum lithium spectrophotometric method provides a useful alternative to ISE or flame photometry, facilitates workstation consolidation on the Hitachi 917 multiple channel analyzer, and is well adapted for use in a rapid response laboratory. PMID- 14749558 TI - Rock on--staying focused on our way to greatness. PMID- 14749559 TI - Trauma and emergency surgery: an evolutionary direction for trauma surgeons. AB - BACKGROUND: The success of nonoperative management of injuries has diminished the operative experience of trauma surgeons. To enhance operative experience, our trauma surgeons began caring for all general surgery emergencies. Our objective was to characterize and compare the experience of our trauma surgeons with that of our general surgeons. METHODS: We reviewed records to determine case diversity, complexity, time of operation, need for intensive care unit care, and payor mix for patients treated by the trauma and emergency surgery (TES) surgeons and elective practice general surgery (ELEC) surgeons over a 1-year period. RESULTS: TES and ELEC surgeons performed 253 +/- 83 and 234 +/- 40 operations per surgeon, respectively (p = 0.59). TES surgeons admitted more patients and performed more after-hours operations than their ELEC colleagues. Both groups had a mix of cases that was diverse and complex. CONCLUSION: Combining the care of patients with trauma and general surgery emergencies resulted in a breadth and scope of practice for TES surgeons that compared well with that of ELEC surgeons. PMID- 14749560 TI - Delayed repair for blunt thoracic aortic injury: is it really equivalent to early repair? AB - BACKGROUND: Blunt thoracic aortic injury (BTAI) is a severe injury that traditionally has mandated immediate surgical repair. Delaying operative intervention for BTAI can allow other life-threatening injuries to be managed first, but potentially increases the risk of aortic rupture and death. The objective of this study was to evaluate the outcome of delayed repair (DR) compared with early repair (ER) for BTAI and to assess the effectiveness of a protocol for medical control of systolic blood pressure and heart rate in those patients whose repairs were delayed. METHODS: This study is a retrospective review of University of Michigan Health System (UMHS) data from January 1, 1992, through March 1, 2003. ER was defined as operative repair within 16 hours from the time of injury. A similar analysis was conducted for patients with BTAI selected from the National Trauma Data Bank. RESULTS: For the UMHS data, there were 45 patients in the DR group and 33 patients in the ER group. Mortality in the ER group versus the DR group was 9% versus 20%. Multivariate analysis adjusting for age, Injury Severity Score, abdominal Abbreviated Injury Scale score, Glasgow Coma Scale score, and intubation status demonstrated an odds ratio for death from ER compared with DR of 1.72 (p = 0.57). Patients undergoing DR had an absolute increase in hospital length of stay (33.1 vs. 20.9 days) and complication rate (2.1 vs. 1.5 incidents per patient). A similar result was obtained for multivariate analysis of the National Trauma Data Bank data, with an odds ratio of 1.40 (p = 0.51) for death from ER versus DR. UMHS patients whose repairs were delayed achieved target systolic blood pressure and heart rate for 76% and 74% of the hourly measurements recorded, respectively. CONCLUSION: Patients with BTAI can safely undergo delayed aortic repair if other injuries warrant a higher treatment priority without increasing their overall risk of mortality. Delayed repair is, however, associated with a higher complication rate. PMID- 14749561 TI - Legal prosecution of alcohol-impaired drivers admitted to a level I trauma center in Rhode Island. AB - BACKGROUND: Despite harsh legislation, driving under the influence of alcohol (DUI) is exceedingly common, and alcohol-related motor vehicle crashes (MVCs) account for significant morbidity, mortality, and economic loss. Legal sanctions can keep offenders off the road (protecting other drivers and pedestrians) and ensure compliance with treatment programs (reducing recidivism). However, even with clear evidence of a transgression, the law seems to be inconsistently enforced among trauma patients. The purpose of this study was to measure the rate of legal prosecution among impaired drivers admitted to a trauma center after MVCs, and to determine the recidivism rate among these individuals. METHODS: Our trauma registry was queried to identify intoxicated drivers admitted during an 18 month period. Court records identified patients who had been charged with traffic offenses, including prior (2 years) and subsequent (1 year) charges. RESULTS: Blood alcohol concentration (BAC) was measured in 387 (74%) of 525 drivers, of whom 137 (35%) had BAC > or =100 mg/dL. Of 113 state residents, 22 (19%) were charged with an offense related to the MVC. Of 12 charged with DUI, 10 were convicted, for an overall DUI conviction rate of just 9%. Seven (32%) of those who were charged had prior or subsequent charges. Of 91 patients not charged for the index event, 31 (34%) had prior or subsequent charges. CONCLUSION: Alcohol is involved in a large percentage of MVCs in our region. The infrequency of prosecution for DUI despite property damage and/or personal injury, and the high recidivism rate, are significant social concerns. These data suggest the need for processes to facilitate legal prosecution--possibly including revision of legislation involving reporting of BACs. PMID- 14749562 TI - Outcome of tracheobronchial injuries: a long-term perspective. AB - BACKGROUND: Tracheobronchial injuries are relatively uncommon, and few data are available on the long-term effects of their treatment. METHODS: All injuries involving the larynx and trachea, trachea alone, and mainstem bronchus (MSB) treated by one surgeon were followed if they survived 48 hours. RESULTS: Sixty patients were treated from 1976 to 2001 for blunt and penetrating injuries: 6 laryngotracheal injuries, 27 tracheal wounds, and 27 injuries to the mainstem bronchus. Follow-up ranged from 1 to 26 years. One of six laryngotracheal wounds had a good result. One required tracheal resection and one required permanent tracheostomy. Patients who survived tracheal resection and end-to-end anastomosis had good outcomes; two had granulomata caused by permanent suture use. One patient treated by primary repair developed stenosis requiring resection. Fourteen patients with MSB injury were treated by pneumonectomy, eight of whom survived. Three developed stump leak/empyema and three had cor pulmonale on long term follow-up. Ten patients had repair of blunt MSB injuries; two developed bronchial stenosis requiring pneumonectomy. CONCLUSION: Laryngotracheal and MSB injuries often had less than optimal outcomes on long-term observation. Tracheal injuries treated by resection and end-to-end repairs had excellent outcomes. The data should be useful in counseling patients/families and planning follow-up strategies for patients with tracheobronchial injuries. PMID- 14749563 TI - The long-term effect of oxandrolone on hepatic acute phase proteins in severely burned children. AB - BACKGROUND: Acute phase protein production is a hallmark of severe burns. We wondered whether anabolic treatment with oxandrolone would affect these proteins. METHODS: Thirty-five children with > or =40% total body surface area burns were randomized to receive either placebo or oxandrolone (0.1 mg/kg by mouth twice daily) from postoperative day 5 to 1 year postburn. Levels of constitutive proteins and acute phase proteins were measured at admission; at discharge; and at 6, 9, and 12 months after burn. Total albumin supplementation and hepatic transaminases were also assessed. RESULTS: Constitutive proteins such as albumin, prealbumin, and retinol-binding protein levels increased (p < 0.05), and acute phase proteins such as alpha 1-acid glycoprotein, C3 complement, alpha 2 macroglobulin, and fibrinogen levels significantly decreased in the oxandrolone group compared with placebo (p < 0.05). Albumin supplementation during the acute hospitalization was reduced in the oxandrolone group. Hepatic transaminases remained within normal levels. CONCLUSION: Treatment with oxandrolone in severe burns significantly increases constitutive protein and reduces acute phase protein levels. PMID- 14749564 TI - Patterns of microbiology in intra-abdominal packing for trauma. AB - INTRODUCTION: This study tracks the microbiology of packs and infections in damage-control trauma patients to determine whether the packs cause infections. METHODS: The peritoneum and abdominal packs were cultured in patients who survived to re-operation. The study recorded all positive cultures, pack count, packing duration, number of operations, and infections. RESULTS: Thirty-five patients were studied. Twenty-eight patients survived; seven died. Packs were cultured in 29 patients. Data for 291 cultures collected. Pack cultures were positive in 20 patients and negative in nine. Positive pack cultures grew skin and gut flora. Twenty-one patients had infections, 14 did not. Organisms from positive pack cultures did not contribute to subsequent infections or mortality. Microbes and sites of infections were consistent with SICU patients. CONCLUSIONS: Intra-abdominal packs are contaminated with skin and gut flora. These contaminants, however, do not contribute to subsequent infections. Pathogens from subsequent infections were typical for ICU infections. PMID- 14749565 TI - Early surgery for thoracolumbar spine injuries decreases complications. AB - BACKGROUND: The proper timing for surgical fracture repair is controversial. Early repair of long bone and cervical fractures reduces complications and is safe. Few studies exist to compare time to surgery with outcomes in thoracolumbar (TL) spine injuries. METHODS: Patients with TL spine injuries were identified from the trauma registry and divided into two cohorts on the basis of Injury Severity Score (ISS). Cohorts were compared for infectious, respiratory, and total complications in patients who had early (<72 hours from injury) versus late (>72 hours from injury) surgical repair. A retrospective chart review was performed on High ISS patients (> or =15) to identify differences in resuscitation needs and neurologic, respiratory, and infectious complications. RESULTS: Early surgery, Low ISS patients were younger, received fewer anterior repairs, and had shorter hospitalizations. Early patients in the High ISS cohort had significantly fewer total complications and shorter hospital and intensive care unit lengths of stay. Resuscitative requirements were similar for both surgery groups. More late surgery patients required ventilator support for noninfectious reasons. There was no difference in admission or postoperative neurologic status or the incidence of head injury. CONCLUSION: Early surgery in severely injured patients with thoracolumbar spine trauma was associated with fewer complications and shorter hospital and intensive care unit lengths of stay, required less ventilator support for noninfectious reasons, and did not increase neurologic deficits. PMID- 14749566 TI - Falling down and falling out: management and outcome analysis. AB - BACKGROUND: This study compares young (< 65 years old) and elderly (> or = 65 years old) patients who fall secondary to syncope and seeks to determine whether syncope workups are being appropriately performed and whether they contribute new information that results in a change in management. METHODS: A retrospective review of patients who fell and were admitted to a Level I trauma center was performed. Data included mechanism of injury, comorbidities, and severity scores in addition to details regarding a syncope workup in patients who had unclear reasons for falling. Outcome variables were mortality, intensive care unit and hospital lengths of stay, and whether each test resulted in a change in management. RESULTS: The data set included 387 patients. Elderly patients who fell (n = 157) had significantly higher Injury Severity Scores and mortality, lower Glasgow Coma Scale scores, and longer intensive care unit and hospital lengths of stay than the younger cohort (n = 230). When a fall occurred secondary to syncope, however, there was no difference in injury severity or outcome. Patients who fell secondary to syncope (n = 61) had zero to six of the recommended tests ordered. Nineteen tests in the young group and 79 tests in the elderly patients had abnormal results. Overall, 37.8% of patients had specific interventions performed because of the abnormal test results CONCLUSION: Syncope workups were erratically performed in both young and older groups. These workups frequently resulted in abnormal findings that required intervention. Protocols are currently being developed at our institution to ensure complete assessment of trauma patients who fall for unknown reasons. PMID- 14749567 TI - Specific occupant and collision characteristics are associated with motor vehicle collision-related blunt cerebrovascular artery injury. AB - BACKGROUND: Blunt cerebrovascular artery injury (BCI) remains difficult to diagnose but is recognized with increasing frequency after motor vehicle collisions (MVCs). Failure to detect this injury in a timely fashion can be devastating. Criteria that can be used to heighten the suspicion of this injury have been suggested; however, more encompassing screening has been recommended. To address this need, we sought to describe occupant, vehicle, and collision characteristics among MVC occupants who sustained a BCI. METHODS: All cases of BCI identified in the National Automotive Sampling System Crashworthiness Data System, a national probability sample of passenger vehicles involved in police reported tow-away MVCs, between 1993 and 2001 were selected. Information on occupant (e.g., demographics, seating position, and restraint use), collision (e.g., collision type and severity), and vehicle characteristics were obtained and summarized using descriptive statistics. RESULTS: Nine-hundred forty individuals with BCI were identified in the Crashworthiness Data System data files. Over half were belted (57.4%) and 82.3% had airbag deployment; 16.2% were partially or completely ejected from the vehicle. Head and thoracic injuries were common (44.4% and 40.8%, respectively); 27.8% sustained a cervical spine fracture and 21.0% sustained a soft-tissue injury to the neck. The mean Injury Severity Score was 33.6. The case fatality rate was 44.5%. The majority of BCI occupants were drivers (76.0%). Among belted occupants, the lap/shoulder was the most commonly attributed as the injury source (61.4%). Among unbelted occupants, frequent injury sources included air bags (15.0%), windshield (13.7%), and other interior objects. With respect to collision characteristics, the average change in velocity (Delta V) was 43.3 km/h. The majority of collisions were frontal (76.2%). CONCLUSION: This study indicates that BCI is both a rare and lethal injury typified by specific occupant and collision characteristics. These characteristics provide insight as to the cause of this injury that may aid in the evaluation and management of the blunt trauma patient at risk for BCI. PMID- 14749568 TI - The epidemiology of serious and fatal injury in San Diego County over an 11-year period. AB - BACKGROUND: Analysis of the mechanism and severity of injury over time may permit a more focused planning of acute care and trauma prevention programs. METHODS: A retrospective, population-based study examining severe traumatic injury in a single county was undertaken. Three overlapping data sets were used to form a composite injury data set. RESULTS: There were 55,664 patients included in the study. A total of 40,897 (73.5%) patients survived and 14,767 (26.5%) died. Of those patients who died, 8,910 (60.3%) died in the field and were not transported to a trauma center. There was an increase in the mean age of all trauma victims (3 years) and an increase of 5 years in fatally injured patients. The mean Injury Severity Score decreased from 14.7 to 11.6 (p < 0.01); however, Injury Severity Score for fatal patients remained constant (39.7). The overall injury rate remained unchanged (195 per 10(5)), whereas the fatal injury rate decreased by 22% (45.9 per 10(5)) over the 11-year study period. The leading cause of injury was motor vehicle crash, followed by assault. The leading cause of fatal injury was suicide, followed by homicide. CONCLUSION: A combination of three independent injury data sources generated a composite data set of serious and fatal injury. This regional injury analysis was the most comprehensive overview of injury in our region. Important observations included the following: there has been no change in the overall incidence of severe injury within our county; the incidence of fatal traumatic injury has significantly decreased; the leading causes of nonfatal injury do not correlate with the rank order of fatal injury; intentional injury was the leading cause of injury deaths; and scene fatalities represent a poorly studied group of patients who may benefit from primary prevention and injury control research. PMID- 14749569 TI - The combination of platelet-enriched autologous plasma with bovine collagen and thrombin decreases the need for multiple blood transfusions in trauma patients with retroperitoneal bleeding. AB - OBJECTIVES: Bleeding from blunt and penetrating retroperitoneal injuries during operative exploration are often difficult to control surgically and can be associated with significant blood loss. Our goals were to evaluate and compare the efficacy of a topical autologous platelet-enriched plasma combined with bovine collagen and thrombin (PCT) to Gelfoam/thrombin (G/T) in relation to hemostatic control/blood transfusion (BTx) requirements and subsequent outcome. METHODS: Prospective data were collected on all patients who underwent operative exploration for retroperitoneal injuries in which either PCT or G/T was applied with or without packing over a 2.5-year period. Patients were stratified by age, gender, mechanism of injury, preoperative international normalized ratio, pH, hematocrit, intraoperative blood loss, and BTx requirements. Subsequent BTx requirements were calculated within 48 hours of the surgical procedure. Outcome was measured by intensive care unit and hospital length of stay and mortality. RESULTS: A total of 78 patients met study criteria. Patients who received G/T had a significantly greater number of early postoperative transfusions (p < 0.001) and a longer hospital (p < 0.001) and intensive care unit length of stay (p < 0.007). There was no difference in mortality. CONCLUSION: PCT is a rapidly available topical hemostat that is associated with a significant decrease in the need for postoperative blood transfusions and intensive care unit and hospital length of stay. A randomized prospective trial to confirm these results is warranted. PMID- 14749570 TI - Blunt diaphragmatic rupture in children. AB - BACKGROUND: Although several series of blunt diaphragmatic rupture in adults have been published, this injury remains largely uncharacterized in the pediatric population. METHODS: We queried our trauma registry for all children admitted with blunt diaphragmatic rupture over a 10-year period at a Level I pediatric trauma center. RESULTS: Six children (aged 2-15 years; mean, 7 years) were identified with blunt diaphragmatic rupture (three right, two left, one bilateral), representing 0.4% of admissions. All of the children had associated injuries (4.5 per child), with a mean Injury Severity Score of 32. Four diaphragmatic injuries were identified during the initial evaluation. The two missed injuries were diagnosed at postinjury days 5 and 8. There were no deaths and all children were eventually discharged without sequelae. CONCLUSION: Blunt diaphragmatic rupture occurs in children with a frequency and severity commensurate with that observed in adults. Our data suggest improved survival compared with adults with this injury. PMID- 14749571 TI - Pelvic fractures and associated injuries in children. AB - BACKGROUND: Pelvic fractures occur uncommonly in children. Despite serious sequelae, they have been infrequently reviewed. METHODS: We conducted a retrospective review of admissions to our institution from January 1983 to December 2000. RESULTS: One hundred twenty children with pelvic fractures were identified. Median age was 9 years (range, 1-16 years) and 66% (n = 80) were boys. Pedestrian-motor vehicle injury accounted for 68% (n = 82) of cases. Associated injuries were present in 78% (n = 94). Management of the pelvic fracture was nonoperative in 113 (94%). Thirty-two children (27%) required surgery for associated injuries. Complications during admission occurred in 28% (n = 34). Five children died as a result of their injuries. With a mean follow-up of 36 months (range, 7-156 months), 27% (n = 32) of children suffered an adverse outcome, including neurologic dysfunction and leg-length discrepancies. CONCLUSION: The majority of pelvic fractures in children may be satisfactorily treated nonoperatively. Operative interventions were more frequently required for associated injuries. Long-term review is indicated because of delayed complications in children that are continuing to grow and develop. PMID- 14749573 TI - Impact of helicopter transport and hospital level on mortality of polytrauma patients. AB - BACKGROUND: Despite numerous studies analyzing this topic, specific advantages of helicopter transport of blunt polytrauma patients as compared with ground ambulances have not yet been identified unequivocally. METHODS: Four possible pathways in 403 polytrauma patients (Injury Severity Score [ISS] > 16) who were in reach of the helicopter emergency medical service (HEMS) Dresden were analyzed as follows: HEMS-UNI group (n = 140), transfer by HEMS into a university hospital; AMB-REG group (n = 102), transfer by ground ambulance into a regional (Level II or III) hospital; AMB-UNI group (n = 70), transfer by ground ambulance into the university hospital; and INTER group (n = 91), transfer by ground ambulance into a regional hospital, followed by transfer to the university hospital. Scores used were the ISS and the TRISS. Tests used for statistical analysis included chi2 and Fisher's tests. Statistical significance was set at p > 0.05. RESULTS: Age, gender, and mean ISS (range, 33.3-35.6) revealed extensive homogeneity of the groups. Mortality of the AMB-REG group was almost doubled (41.2%) compared with HEMS-UNI (22.1%) patients (p = 0.002). The AMB-UNI group displayed the lowest mortality (15.7%, p = not significant). TRISS analysis (PRE Chart) revealed identical outcome for AMB-UNI and HEMS-UNI patients. Rescue time averaged 90 +/- 29 minutes for HEMS-UNI patients, 68 +/- 25 minutes for AMB-UNI patients, and 69 +/- 26 minutes for the AMB-REG group. CONCLUSION: Primary transfer by HEMS into a Level I trauma center reduces mortality markedly. In principle, this benefit can be attributed to superior preclinical therapy, primary admission to a Level I trauma center, or both. However, the identical probability of survival of the AMB-UNI and HEMS-UNI groups in this and comparable studies does not confirm generally better survival rates on account of a more aggressive on-site approach. PMID- 14749572 TI - Effective use of the air ambulance for pediatric trauma. AB - BACKGROUND: The purpose of this study was to compare outcomes of pediatric trauma patients transported by helicopter from the injury scene (IS group) to a trauma center and those transported by air after hospital stabilization (HS group). METHODS: A retrospective analysis of pediatric trauma patients (<19 years of age) transported by air ambulance and admitted to a pediatric trauma center was conducted. Outcomes compared were mortality and length of stay. Patients were subdivided into minor (Injury Severity Score [ISS] < 15) and major (ISS > 15) trauma. TRISS analysis was performed to verify the overall quality of the care. RESULTS: Eight hundred forty-two HS and 379 IS patients were included. The mean age, median ISS, and distribution of penetrating and blunt injuries did not differ significantly between the groups. The overall death rate was significantly lower for the interfacility transfer patients (HS group, 5.5%; IS group, 8.7%; p < 0.05). Mean intensive care unit (ICU) and hospital length of stay did not differ significantly. HS patients with major trauma had significantly less mortality (HS group, 15.5%; IS group, 26.7%; p < 0.05) and shorter mean ICU stays (HS group, 118.3 hours; IS group, 149.1 hours; p < 0.05) than IS major trauma patients. No differences were seen in patients with minor trauma. TRISS analysis showed improved survival for all patients compared with Major Trauma Outcome Study norms. CONCLUSION: Retrospective analysis was not able to demonstrate any benefit to direct transport from the scene to a trauma center. Hospital stabilization before transfer by air ambulance may improve survival and shorten ICU stays for patients with major trauma. PMID- 14749574 TI - Establishment of a teaching animal model for sonographic diagnosis of trauma. AB - BACKGROUND: Ultrasound is widely accepted as a valuable diagnostic tool for detecting intra-abdominal and intrathoracic bleeding in trauma patients. Nevertheless, many doctors are reluctant to use it because they do not have sufficient training. This study aimed to define intra-abdominal and intrathoracic fluid volumes that can be detected by sonography and their relation to fluid width in pigs to establish a clinically relevant animal model for teaching and training. METHODS: Different volumes of normal saline were infused into the abdomen (50-2,000 mL) and chest (25-250 mL) in five anesthetized pigs. The maximum width of fluid as detected by ultrasound was recorded. The right upper quadrant, left upper quadrant, pelvis, and right paracolic section of the abdomen and right pleural cavity were studied. An experienced radiologist performed the studies. The effects on respiratory and cardiovascular functions were evaluated. RESULTS: The sonographic findings in the pig were similar to those in humans. Up to 50 mL of intra-abdominal fluid and up to 25 mL of intrathoracic fluid could be detected by ultrasound. There was a significant correlation between the volume infused and the fluid width detected. The respiratory and cardiovascular monitoring of the animals showed that the infused intrathoracic volumes mimicked a survivable hemothorax. CONCLUSION: The pig may serve as an excellent clinically relevant model with which to teach surgeons detection of different volumes of intra-abdominal and intrathoracic fluids. The value of this model as an educational tool has yet to be tested. PMID- 14749575 TI - Intestinal bacterial overgrowth induces the production of biologically active intestinal lymph. AB - OBJECTIVE: We have previously documented that gut-derived lymph from rats subjected to trauma plus hemorrhagic shock (T/HS) is injurious to vascular endothelial cells and activates neutrophils (PMNs), two key events in postshock organ injury. Because T/HS leads to gut injury, intestinal bacterial overgrowth, and the loss of gut barrier function, the relative role of gut injury as opposed to intestinal bacterial overgrowth per se in the pathogenesis of biologically active intestinal lymph is unclear. We therefore studied whether mesenteric lymph can injure endothelial cells and/or active PMNs in an intestinal bacterial overgrowth model where there is no gut injury (monoassociation). METHODS: Bacterial overgrowth was established in male rats by treating the animals with 4 days of oral antibiotics followed by administration of a nonpathogenic, streptomycin-resistant strain of Escherichia coli C25. Mesenteric lymph was then collected from rats with normal flora and from E. coli C25 monoassociated rats. Its effects were tested on human umbilical vein endothelial cells (HUVECs) and human PMNs. As an additional control, lymph was collected from antibiotic decontaminated rats that received antibiotics but were not colonized with E. coli C25. RESULTS: As compared with medium, normal flora intestinal lymph, antibiotic decontaminated lymph, or portal plasma from the monoassociated rats, mesenteric lymph from the monoassociated rats killed HUVECs and increased the permeability of a HUVEC monolayer. In contrast to the effects on HUVECs, lymph from the monoassociated rats did not increase PMN CD11b expression or prime PMNs for an augmented respiratory burst, as compared with lymph from the rats with normal flora or from antibiotic-decontaminated rats. The effects of lymph from the monoassociated rats was not caused by bacteria, because these lymph samples were sterile. CONCLUSION: These results indicate that disruption of the normal intestinal microflora resulting in bacterial overgrowth with enteric bacilli may participate in the production of mesenteric lymph that is injurious to endothelial cells in shock, but this mechanism does not appear to be significantly involved in the activation of PMNs. PMID- 14749576 TI - Imipenem levels are not predictable in the critically ill patient. AB - BACKGROUND: Critically ill patients often demonstrate extremely unusual volumes of distribution (Vd) and half-lives (t1/2) of drugs. Imipenem is a widely used antibiotic in critically ill patients. METHODS: We performed high-performance liquid chromatography analysis of imipenem in samples from 50 critically ill patients treated with either 500 or 1,000 mg. RESULTS: Peak imipenem levels varied from 1.56 microg/mL to 58.8 microg/mL. Trough levels varied between 0.0 microg/mL and 15.62 microg/mL. Only 54% of patients maintained a trough level greater than 4 microg/mL. Both the Vd and the t1/2 of imipenem were much greater than observed in other patient populations. CONCLUSION: The pharmacokinetic activity of imipenem in critically ill patients is different from that in other patient populations. There is a very weak correlation between dosage and serum concentrations. Therapeutic failures of imipenem may be because of unpredictable pharmacodynamics (Vd and t1/2) in critically ill surgical patients. PMID- 14749577 TI - Alterations in resistin expression after thermal injury. AB - BACKGROUND: Burn injury, it was hypothesized, may induce changes in resistin expression that contribute to postburn metabolic derangements. This study examined resistin gene expression, serum levels of resistin protein, and glucose levels in burned mice. METHODS: Ten male Balb-c mice were anesthetized and then given a 30% total burn surface area using heated probes. Burned and sham-burned mice were killed at 2, 4, 24, and 48 hours. The total ribonucleic acid from gonadal fat tissues was isolated for the measurement of resistin gene expression using real-time reverse transcriptase-polymerase chain reaction. Serum levels of resistin, insulin, and glucose were measured. Statistical analysis was performed by two-way analysis of variance using Bonferroni's test to find differences between groups. All p values less than 0.05 were considered significant. RESULTS: Increases in resistin gene expression and serum resistin levels were detected in the burned animals, and these correlated with relative insulin resistance. CONCLUSION: The findings suggest a potential role for resistin in the pathophysiology of the metabolic response to injury. PMID- 14749578 TI - Intravenous catheter complications in the hand and forearm. AB - BACKGROUND: We studied the complications of peripheral intravenous (i.v.) catheters in the hand and forearm in a teaching hospital over a 3-year period. METHODS: The records of 67 patients who developed i.v. catheter-related complications were reviewed. RESULTS: The most common sites for developing complications in order of frequency were the forearm, hand, wrist, and antecubital fossa. There were 56 minor and 11 major complications. More than 50% of minor complications occurred in the hand and wrist, and more than 50% of major complications occurred in the hand. In 68% of minor complications, the patients were aged 50 years or older and 68% were women. Minor complications comprised 26 intravenous infiltrations, 23 cases of thrombophlebitis, and 7 cases of cellulitis. Ninety percent of major complication patients were aged 50 or older and 82% were women. Major complications included septic thrombophlebitis in three; hematomas resulting in skin necrosis in two; and infiltration related complications in six, resulting in skin necrosis in two, compressive nerve lesions in two, digital stiffness in one, and compartment syndrome in one. Ten patients with major complications were over the age of 50 years and nine were women. Two patients receiving anticoagulation developed large dorsal subcutaneous space hematomas. Chemotherapeutic agents contributed to two minor complications and one major complication. CONCLUSION: The hand is a common site for minor and major i.v. catheter complications. Women and older patients are more susceptible to these complications. Peripheral i.v. line complications are not uncommon and can result in morbidity and increased health care costs from prolonged hospitalization, extended use of i.v. antibiotic therapy, and surgical intervention. PMID- 14749579 TI - The death of articular chondrocytes after intra-articular fracture in humans. AB - BACKGROUND: In this study, we wished to determine whether chondrocyte death occurs after intra-articular fracture and might thus contribute to the development of posttraumatic arthritis. METHODS: Articular fracture fragments were obtained from 30 patients and the terminal deoxynucleotide transferase mediated dUTP nick-end labeling assay was used to identify dead or dying cells. RESULTS: Dead or dying cells were identified in all fracture fragments, with an average rate of cell death of 35%. Rates of cell death greater than 90% were seen in 4 of the 30 specimens studied. CONCLUSION: Although rates of apoptosis in osteoarthritis have been reported to average 15%, the rate in this fracture population was more than twice this value (35%). The high rate of cell death noted here may help explain the occurrence of posttraumatic arthritis even in anatomically fixed intra-articular fractures. PMID- 14749580 TI - Bedside fluoroscopic flexion and extension cervical spine radiographs for clearance of the cervical spine in comatose trauma patients. AB - BACKGROUND: Bedside flexion and extension fluoroscopic examinations have been proposed as an option for clearance of the cervical spine in comatose brain injured patients. We hypothesized that these studies, when performed after normal static imaging of the cervical spine, would have an extremely low likelihood of identifying occult ligamentous instability and would not be adequate for visualizing the lower cervical spine. METHODS: Radiographic images obtained from 56 consecutive comatose head-injured patients were reviewed. All patients had normal anteroposterior, lateral, and open mouth odontoid cervical spine radiographs and normal thin-cut axial computed tomographic images from the occiput to C2 and through the lower cervical spine if suspicious areas were identified on plain cervical spine radiographs. After these static images were determined to be normal by both the attending neurosurgeon and the attending radiologist, all 56 patients had bedside fluoroscopic flexion and extension studies performed by the neurosurgery resident, with the patients' arms being pulled down to their sides by the primary care nurse. RESULTS: The bedside fluoroscopic flexion and extension studies were considered to be adequate (visualization to the C7-T1 motion segment) in only 4% of the patients. Occult instability was identified in one patient (type II odontoid fracture) and significant instability was missed in one patient with C6 to C7 dislocation in whom flexion and extension radiographs failed to visualize the C6 to C7 motion segment. CONCLUSION: Bedside flexion and extension fluoroscopy was almost always inadequate for visualizing the lower cervical spine in comatose head-injured patients. Because of the extremely low likelihood of visualizing the entire cervical spine with this technique, we recommend that it no longer be considered an option in trauma center protocols for clearance of the cervical spine in comatose brain-injured patients. PMID- 14749581 TI - Evaluation of management of road trauma survivors with brain injury and neurologic disability in Victoria. AB - BACKGROUND: Victoria recently established a new trauma care system following the Consultative Committee's findings on frequent preventable deaths after road crash injury. This study investigates the contribution to neurologic disability of preventable deficiencies in health care in survivors of road crashes occurring from 1998 to 1999. METHODS: The emergency and clinical management of 60 road crash survivors with head Abbreviated Injury Scale score > or = 3 and residual neurologic disability were evaluated by analysis and multidisciplinary discussion of their complete prehospital, hospital, and rehabilitation records. RESULTS: The mean number of potentially preventable errors or inadequacies per patient was 19.2 +/- 7.5, with 10.5 +/- 7.2 contributing to neurologic disability. The mean number contributing to neurologic disability was greatest in the emergency room (3.5 +/- 3.2), followed by the intensive care unit (2.2 +/- 2.7) and the prehospital setting (1.8 +/- 2.0). Eighty-four percent of the deficiencies were management errors/inadequacies and 7% were system inadequacies. Fifty-five percent of deficiencies contributed to neurologic disability. In patients with a systolic blood pressure less than 90 mm Hg with hypovolemia consequent to inadequate resuscitation, the frequency of severe neurologic disability was increased almost twofold (p < 0.05). Deficiencies contributing to neurologic disability were significantly less frequent in university teaching hospitals with neurosurgical units. CONCLUSION: Improvement in neurologic outcomes can be achieved through appropriate triage and increased attention to basic principles of trauma and head injury care. PMID- 14749582 TI - Distribution and determinants of health and work status in a comprehensive population of injury patients. AB - BACKGROUND: Insight into the distribution and determinants of both short- and long-term disability can be used to prioritize the development of prevention policies and to improve trauma care. We report on a large follow-up study in a comprehensive population of injury patients. METHODS: We fielded a postal questionnaire in a stratified sample of 4,639 nonhospitalized and hospitalized injury patients aged 15 years and older, at 2, 5, and 9 months after injury. We gathered sociodemographic information, data on functional outcome with a generic instrument for health status measurement (EuroQol EQ-5D+) and data on work absence. RESULTS: The response rates were 39%, 75%, and 68% after 2, 5, and 9 months, respectively. The reported data were adjusted for response bias and stratification. The 2-month health status of nonhospitalized patients was comparable to the general population's health when measured by the EQ-5D summary score, although considerable prevalence of restrictions in usual activities (24.0%) and pain and discomfort (34.8%) were reported. Hospitalized patients reported higher prevalences of disability in all health domains. Their mean EQ-5D summary score increased from 0.62 at 2 months to 0.74 at 5 months but remained below the population norm at 9 months, particularly for patients with a long hospital stay. Patients with injuries of the spinal cord and vertebral column, hip fracture, and other lower extremity fractures reported the worst health status, also when adjusted for age, sex, and educational level. Age, sex, type of injury, length of stay, educational level, motor vehicle injury, medical operation, intensive care unit admission, and number of injuries were all significant predictors of functioning. Nonhospitalized and hospitalized injury patients lost on average 5.2 and 72.1 work days, respectively. Of nonhospitalized patients, 5% had not yet returned to work after 2 months, and 39%, 20%, and 10% of hospitalized patients had not yet returned to work after 2, 5, and 9 months, respectively. In a multivariate regression analysis, length of stay, type of injury, level of education, and intensive care unit admission appeared to be significant predictors of absence duration and return to work. CONCLUSION: Injury is a major source of disease burden and work absence. Both hospitalized and nonhospitalized patients contribute significantly to this burden. PMID- 14749583 TI - Comparison of the New Injury Severity Score and the Injury Severity Score. AB - BACKGROUND: The New Injury Severity Score (NISS) was proposed in 1997 to replace the Injury Severity Score (ISS) because it is more sensitive for mortality. We aim to test whether this is true in our patients. METHODS: This study was a retrospective review of data from 6,231 consecutive patients over 3 years in the trauma registry of a Level I trauma center studying outcome, ISS, and NISS. RESULTS: Misclassification rates were 3.97% for the NISS and 4.35% for the ISS. The receiver operating characteristic curve areas were 0.936 and 0.94, respectively. Neither the ISS nor the NISS were well calibrated (Hosmer-Lemeshow statistic, 36.11 and 49.28, respectively; p < 0.001). CONCLUSION: The NISS should not replace the ISS, as they share similar accuracy and calibration. PMID- 14749585 TI - A resource-based assessment of trauma care in the United States. AB - BACKGROUND: The resources needed and those available to support trauma care for a given region are currently unknown. Resource use and availability were evaluated for injured subjects across a large sample of the United States. METHODS: This population-based study of trauma-related discharges in 18 states represented all four geographic regions of the United States. Hospital discharge and bed utilization rates as a function of injury severity were assessed. Resource availability was evaluated by determining state trauma center density. RESULTS: This study evaluated 523,780 trauma patients discharged from 2,317 hospitals. The discharge rate for all trauma was 412 per 100,000 person-years, whereas the rate for major trauma was only 44 per 100,000 person-years. More than one third of the patients with major trauma received care at centers not designated for trauma care. The hospital bed utilization rate was 2,095 days per 100,000 person-years. The availability of trauma centers varied greatly across states, ranging from 0.9 to 6.6 centers per million population. CONCLUSIONS: A substantial minority of major trauma patients in the United States are treated in nondesignated trauma centers. The variability in the availability of trauma resources indicates a lack of consensus with respect to the resources required for trauma system implementation. PMID- 14749584 TI - Judging trauma center quality: does it depend on the choice of outcomes? AB - BACKGROUND: Trauma centers routinely benchmark their survival outcomes against a national norm using the TRISS methodology. However, the use of survival as a measure of the effectiveness of trauma care may be too limited in scope because it fails to capture information regarding functional outcomes. METHODS: The objective of this study was to develop a prediction model that allows hospitals to benchmark their functional outcomes in blunt trauma patients, and to determine whether the assessment of hospital "quality" depends on the choice of outcome measure: survival or survival combined with functional outcome. This retrospective cohort study was based on patients, aged 18 years or older, in the National Trauma Database who sustained blunt trauma in 1999 without associated head or spinal cord injury. We developed a sequential logistic model to predict the probability of a good functional outcome. The TRISS methodology was customized to this data set to obtain a survival model. Using each of these prediction models, we then obtained two standardized measures of hospital performance: one based on the number of survivors and the other based on the number of survivors with good functional outcomes. These standardized outcome measures were then used to identify low-performance and high-performance hospitals. The ranking based on these two different measures were compared. RESULTS: Fifteen of the 27 hospitals in the study cohort were categorized differently when their performance was benchmarked using survival versus functional outcome. Kappa analysis revealed minimal agreement between these two quality measures on the identity of hospital quality outliers (kappa = 0.04; p = 0.35). CONCLUSION: The evaluation of hospital quality depends on whether hospital performance is judged by looking at survival or at survival combined with functional outcome. Because functional status is an important outcome of major concern to survivors, it is important to include it in hospital performance assessment. Consideration should be given to including functional outcome in the evaluation of trauma center performance. PMID- 14749587 TI - Traumatic lumbar nerve root avulsion: evaluation using electrodiagnostic studies and magnetic resonance myelography. PMID- 14749586 TI - Bickerstaff's brain stem encephalitis after a motor vehicle crash. PMID- 14749588 TI - Current management of laryngotracheal trauma: case report and literature review. PMID- 14749589 TI - Blunt avulsion of the right inferior pulmonary vein. PMID- 14749590 TI - Delayed abdominal aortic rupture in a child with a seat-belt sign and review of the literature. PMID- 14749591 TI - Isolated pediatric peripheral vascular injury caused by blunt trauma: a new occurrence. PMID- 14749592 TI - Diagnosis of posttraumatic transverse sinus thrombosis with magnetic resonance imaging/magnetic resonance venography: report of two cases. PMID- 14749594 TI - Restrictive strabismus after blow-out orbital fracture in children: is the muscle involved? PMID- 14749593 TI - Intracorporeal use of the hemostatic agent QuickClot in a coagulopathic patient with combined thoracoabdominal penetrating trauma. PMID- 14749595 TI - Blunt pericardial rupture with cardiac herniation: unusual radiographic findings. PMID- 14749596 TI - Cardiac tamponade and sternal fracture. PMID- 14749597 TI - Prehospital tourniquets: there should be no controversy. PMID- 14749598 TI - Asthma severity and asthma control: symptoms, pulmonary function, and inflammatory markers. AB - PURPOSE OF REVIEW: Asthma severity and asthma control are distinct yet related concepts. Asthma severity describes the underlying disease in the absence of therapy and is ideally defined without concurrent treatment confounding its assessment. Asthma control describes the clinical status of disease in the face of intervention. However, the individual parameters by which we define asthma severity and asthma control overlap significantly. A MEDLINE search between July 1, 2002, and June 30, 2003, was performed using keywords related to concepts of asthma severity of illness and asthma control. From these articles the author selected the articles most relevant for discussion. RECENT FINDINGS: Asthma is a complex syndrome. Although correlations exist between the various parameters used in clinical assessment, no single parameter can accurately classify all individuals. Assessment of multiple parameters including physiologic measures, symptoms, and activity limitation are necessary to categorize asthma clinical status accurately. In addition, the role biomarkers play in the assessment of disease status is an area of increasing interest. Several validated multidimensional measures for assessing asthma control are now available. Each of these measures includes the parameters of symptoms, activity limitation, and rescue medication use, yet they vary on inclusion of other important components such as physiologic measures and biomarkers. SUMMARY: Asthma is a complex syndrome. Currently available instruments demonstrate an improved understanding of the multidimensional approach required to assess asthma control accurately. However, debate continues on the optimal parameters to be included and the role biomarkers play in the clinical assessment of asthma. PMID- 14749600 TI - Macrolide and occult infection in asthma. AB - PURPOSE OF REVIEW: A small percentage of asthma exacerbations are linked with infection by an atypical bacterium, such as Chlamydia pneumoniae or Mycoplasma pneumoniae. These bacteria also have been proposed to cause occult chronic lower airway inflammation and to initiate nonatopic asthma in adults. Consequently, the logical procedure would be to eliminate these pathogens as soon and as thoroughly as possible using antibiotics. Nonetheless, antibiotics are not recommended even for the treatment of acute asthma exacerbations except as needed for comorbid conditions. These discrepancies highlight the need to define the role, if any, of antimicrobials that are active against atypical pathogens, mainly macrolides, but also tetracyclines and fluoroquinolones, in the treatment of asthma. RECENT FINDINGS: Macrolides are antibiotics with both antimicrobial and antiinflammatory activities. Some studies have documented that these agents could be useful in the treatment of occult infection in asthma because of their antimicrobial activity against atypical pathogens. They could also lead to reduction of the airways inflammation by decreasing the transcription of mRNA for a variety of cytokines and inhibiting interleukin-8 release by eosinophils, and therefore improvement of symptoms and pulmonary function. These effects are not caused by bronchodilation, elevation of serum theophylline level, or steroid-sparing mechanism. SUMMARY: The available clinical evidence seems to support use of macrolides in the treatment of asthma because of their antimicrobial activity. However, studies that may confirm this hypothesis are scarce and with limited scientific value because of their open, uncontrolled design. PMID- 14749601 TI - Once-daily inhaled corticosteroids for the treatment of asthma. AB - PURPOSE OF REVIEW: Inhaled corticosteroids (ICS) are the mainstay of asthma therapy. Although compliance to this type of medication is often suboptimal and once-daily dosing can help to improve adherence to the treatment, the clinical implications of such a mode of administration should be determined. RECENT FINDINGS: This review summarizes the recent studies on comparative efficacy of once-versus twice-daily administration of ICS, in light of previous reports. SUMMARY: Although twice-daily administration of ICS is often better to optimize asthma parameters, in many patients, asthma can be sufficiently controlled by a once-daily regimen of most ICS. An increased frequency of dosing seems preferable if asthma becomes uncontrolled or is severe, although this requires further study. A therapeutic trial should, however, be done to ensure that asthma control is adequate. Comparative long-term effects of such a strategy on inflammatory and remodeling parameters remain to be determined, as does the proportion of patients who can adequately control their asthma with once-daily administration of the various ICS available. PMID- 14749602 TI - Recent development in genomic and proteomic research for asthma. AB - PURPOSE OF REVIEW: Asthma is a complex genetic disorder with a heterogeneous phenotype attributed to the interactions among many genes and the environment. This review highlights recent developments in asthma genomic and proteomic research. RECENT FINDINGS: Numerous loci and candidate genes have been reported to show linkage and association of asthma and the asthma-associated phenotypes, atopy, elevated immunoglobulin E (IgE) levels, and bronchial hyperresponsiveness to alleles of microsatellite markers and single nucleotide polymorphisms within specific cytokine/chemokine, and IgE regulating genes. Although many studies reporting these observations are compelling, only a few genes conferring significant risk have been mapped. Although significant progress has been made in the field of asthma genetics in the past decade, the clinical implications of the genetic variations within the numerous candidate asthma genes, which have been found to associate with the expression of the asthmatic phenotype, remain largely undetermined. However, in the past year the scientific community has benefited from postgenomic discoveries, with the recent cloning of two asthma genes, ADAM 33 and PHF11, and this has generated new information that is benefiting others. SUMMARY: The asthma genetics field has advanced considerably in recent years, with new information being generated that has led to improved understanding of the pathobiology underlying this complex disorder. This has also generated interest in the study of gene-gene interaction and how linkage disequilibrium blocks and haplotypes can be used as functional units to pinpoint mutations and capture relative risk of mutated genes in complex disorders. PMID- 14749603 TI - Exhaled nitric oxide in the assessment of asthma. AB - PURPOSE: Asthma is now defined as a TH2-mediated inflammatory disease involving both large and small airways. However, assessment of airways inflammation is limited by techniques that are time consuming and possibly distressing to the patient. Exhaled nitric oxide, an easily and rapidly obtained noninvasive study, is a potential surrogate for measuring airways inflammation, but its clinical utility remains to be determined. This review examines the role of exhaled nitric oxide in assessing and directing therapy of asthmatic airways inflammation. RECENT FINDINGS: It is well established that exhaled nitric oxide is increased in patients with untreated asthma and decreases with corticosteroid treatment. Exhaled nitric oxide also generally correlates with eosinophilic inflammation in asthmatic patients. Recent studies show that this correlation is especially pronounced in atopic subjects with asthma when compared with nonatopic subgroups. Recent studies also show that exhaled nitric oxide may be useful in identifying subclinical inflammation, assessing the antiinflammatory effects of asthma medications other than inhaled or oral corticosteroids, and heralding an asthma exacerbation. A number of new studies assert the utility of exhaled nitric oxide as a diagnostic tool for asthma. SUMMARY: Exhaled nitric oxide may be a useful parameter for monitoring asthmatic inflammation, adjusting therapy, and diagnosing asthma, although prospective longitudinal trials investigating the correlation between exhaled nitric oxide and clinical outcomes are necessary to determine its utility. PMID- 14749604 TI - Complementary and alternative medicine for bronchial asthma: is there new evidence? AB - PURPOSE OF REVIEW: Complementary and alternative medicine is widely used in bronchial asthma. Data on efficacy of these treatment modalities are lacking. RECENT FINDINGS: Studies published since June 2002 on complementary and alternative medicine in bronchial asthma were systematically reviewed. SUMMARY: Studies do not support the use of homeopathy, air ionizers, manual therapy, or acupuncture for asthma. These methods bear some risks to patients related to undertreatment and side effects. There might be a possible, but so far not clearly established, role for antioxidant dietary supplementation, and some natural antiinflammatory and immunomodulatory remedies. However, their effect size compared with the classical treatment and side-effect profile is not clearly established. Strategies influencing breathing technique or perception, such as breathing or retraining exercises, need to be studied over the next few years to establish their additive role in the treatment of asthma. Breathing exercises could improve lung function and quality of life in different studies. Psychotherapy-related methods such as relaxation, hypnosis, autogenic training, speleotherapy, and biofeedback might have a small effect in selected cases, but have not proven to be superior to placebo. Nevertheless, more randomized controlled trials of good methodological quality are required to allow firm conclusions. PMID- 14749605 TI - Clinical phenotypes of asthma. AB - PURPOSE OF REVIEW: Asthma is a phenotypically heterogeneous disorder and, over the years, many different clinical subtypes of asthma have been described. A precise definition of asthma phenotypes is now becoming more and more important, not only for a better understanding of pathophysiologic mechanisms, but in particular to ascertain the specific genes associated with these phenotypes. RECENT FINDINGS: In children, three asthma phenotypes are now well defined: transient infant wheezing, nonatopic wheezing of the toddler, and IgE-mediated wheezing/asthma. Recently, a fourth phenotype, late-onset childhood asthma has been added to this list. In adults, asthma persisting from childhood into adulthood should be distinguished from asthma starting in adulthood. The phenotypes of adult-onset asthma are still poorly defined. Until now, phenotypic classification has been based primarily on etiologic factors (eg, aspirin sensitivity, persistent respiratory infections, occupational factors, or toxic exposures) or clinical characteristics of the disease (eg, mild, severe, brittle, near fatal, with fixed airflow obstruction, steroid resistant). Novel noninvasive techniques to assess the type and severity of airway inflammation and dysfunction are increasingly used to identify better the different phenotypes. SUMMARY: The classic phenotype of IgE-mediated asthma starting in childhood is now clearly defined. However, many other phenotypes of asthma in childhood as well in adulthood are being recognized. In particular, asthma starting in adulthood and noneosinophilic asthma constitute an important part of the adult asthma population, and are still poorly defined. A precise definition of these asthma phenotypes is urgently needed because they are likely to be associated with different genotypes, responses to treatment, and prognoses. PMID- 14749606 TI - Aspirin intolerance and the cyclooxygenase-leukotriene pathways. AB - PURPOSE OF REVIEW: In up to 10% of patients with bronchial asthma, aspirin and other nonsteroidal antiinflammatory drugs precipitate asthmatic attacks. This is a hallmark of a distinct clinical syndrome that develops according to a characteristic sequence of symptoms. Here we discuss its clinical picture and management as related to the abnormalities in arachidonic acid transformations. RECENT FINDINGS: At the biochemical level, the characteristic feature is profound alteration in eicosanoid biosynthesis and metabolism. Major advances in the molecular biology of eicosanoids, exemplified by the cloning of cysteinyl leukotriene receptors and discovery of a whole family of cyclooxygenase enzymes, offer new insights into mechanisms operating in aspirin-induced asthma. Clinical interest has been enhanced by the introduction into therapy of highly specific cyclooxygenase-2 inhibitors and antileukotriene drugs. SUMMARY: Recent studies have improved our understanding of mechanisms operating in asthma and unvieled the role of eicosanoid mediators in pulmonary disease. PMID- 14749608 TI - Outdoor air pollution and asthma. AB - PURPOSE OF REVIEW: The relation between outdoor air pollution and asthma has always been a major focus of research. The evidence that current levels of air pollution in many countries result in increased morbidity and mortality is fairly consistent. With rapid urbanization in many communities, traffic exhausts have become the major source of pollution, and many recent research studies have attempted to investigate the detrimental effects of this type of pollution. This paper reviews the recent evidence of the possible detrimental effects of ambient air pollution on the inception and morbidity of asthma. RECENT FINDINGS: Traffic related pollution has been confirmed in both cross-sectional and longitudinal studies to be associated with increased asthma morbidity and cardiopulmonary mortality. There is also evidence that pollutants such as ozone and traffic exhausts may be responsible for new incident cases of asthma. Among the particulate pollution, research investigating the ultrafine particles and the bacterial components suggested that these particles may have important role in asthma morbidity. SUMMARY: More research studies are needed to reveal how various air pollutants may interact with the host systems, such as the immune system, leading to increased morbidity in susceptible individuals. Reduction of the current levels of ambient air pollution should be an integral part of the overall effort in minimizing asthma morbidity or mortality in the community. PMID- 14749607 TI - Occupational asthma. AB - PURPOSE OF REVIEW: The scientific literature on occupational asthma is steadily increasing and no longer only describes case reports with prevalence figures and limited physiopathologic and immunologic data. Prospective cohort studies are currently carried out with relevant incidence figures and risk factors. Genetic susceptibility is explored. New diagnostic means are described. Surveillance programs are conducted and preventive measures are proposed. RECENT FINDINGS: A relevant account was published on the respiratory consequences (irritant-induced asthma, a type of occupational asthma) of exposure of firefighters at the World Trade Center site. Glutathione-S-transferase appears to be an interesting susceptibility gene for occupational asthma as a result of isocyanates. Inflammation caused by neutrophils and not only by eosinophils is more often associated with occupational asthma. Examination of induced sputum and assessment of exhaled nitric oxide are becoming useful diagnostic tools. Cleaners are at increased risk of developing occupational asthma, although the causal agents are unknown. Surveillance programs currently combine strategies that aim to reduce exposure and identify cases early. SUMMARY: New scientific information provides better, accurate figures on the frequency of occupational asthma and on risk factors, proposes relevant diagnostic tools more directly related to the physiopathology of the disease, and suggests effective surveillance programs in high-risk workplaces. PMID- 14749609 TI - Seasonale. PMID- 14749610 TI - Emergency contraception OTC. PMID- 14749611 TI - Daptomycin (Cubicin) for skin and soft tissue infections. PMID- 14749612 TI - Prevalence of cigarette use among 14 racial/ethnic populations--United States, 1999-2001. AB - The 1998 Surgeon General's report, Tobacco Use Among U.S. Racial/Ethnic Minority Groups, addressed diverse tobacco-control needs of the four primary U.S. racial/ethnic minority populations: non-Hispanic blacks, American Indians/Alaska Natives (AI/ANs), Asians/Pacific Islanders, and Hispanics. However, data on these populations do not describe differences in tobacco-use prevalence among subsets of these populations. To assess the prevalence of cigarette smoking among persons aged > or =12 years among 14 racial/ethnic populations in the United States, CDC analyzed self-reported data collected during 1999-2001 from the National Survey on Drug Use and Health (NSDUH) (formerly the National Household Survey on Drug Abuse). This report summarizes the results of that analysis, which indicated that the prevalence of cigarette smoking among adults aged > or =18 years ranged from 40.4% for AI/ANs to 12.3% for the Chinese population, and the prevalence among youths aged 12-17 years ranged from 27.9% for AI/ANs to 5.2% for the Japanese population. Implementing tobacco-control programs that include culturally appropriate interventions can help reduce tobacco use among racial/ethnic populations. PMID- 14749613 TI - State medicaid coverage for tobacco-dependence treatments--United States, 1994 2002. AB - In 2000, of approximately 32 million persons who received health insurance coverage through Medicaid programs, an estimated 11.5 million (36%) smoked. One of the national health objectives for 2010 is to provide coverage by Medicaid in the 50 states and the District of Columbia (DC) for nicotine-dependence treatment. The Guide to Community Preventive Services recommends reducing the cost of tobacco-dependence treatments to increase the number of smokers who successfully quit smoking. The 2000 Public Health Service (PHS) Clinical Practice Guideline also supports expanded insurance coverage for tobacco-dependence treatments. The amount and type of coverage for tobacco-dependence treatment offered by Medicaid has been reported previously for 1998, 2000, and 2001. In 2002, all states and DC were surveyed again about the amount and type of coverage they provided. This report summarizes the results of the survey, which indicate that as of December 31, 2002, 1) 36 Medicaid programs covered some tobacco dependence counseling or medication for all Medicaid recipients, 2) four states offered coverage only for pregnant women, 3) two states offered coverage for all pharmacotherapy and counseling treatments recommended by the 2000 PHS guideline, and 4) seven states covered all recommended medications and at least one form of counseling. To improve the health of populations with disproportionately high rates of smoking, the 50 states and DC should provide coverage under Medicaid for all recommended tobacco-dependence treatments. PMID- 14749614 TI - Economic costs associated with mental retardation, cerebral palsy, hearing loss, and vision impairment--United States, 2003. AB - Developmental disabilities (DDs) are chronic conditions that initially manifest in persons aged < or =18 years and result in impairment of physical health, mental health, cognition, speech, language, or self-care. The majority of persons with DDs require long-term supportive care or services. In 2003, RTI International (Research Triangle Park, North Carolina) and CDC analyzed data from multiple surveys and reports to estimate the direct and indirect economic costs associated with four DDs in the United States. On the basis of that analysis, estimated lifetime costs in 2003 dollars are expected to total 51.2 billion dollars for persons born in 2000 with mental retardation, 11.5 billion dollars for persons with cerebral palsy, 2.1 billion dollars for persons with hearing loss, and 2.5 billion dollars for persons with vision impairment. These estimates underscore the need for effective primary and secondary prevention measures (e.g., newborn screening for hearing and metabolic disorders and smoking cessation counseling for pregnant women) to reduce the costs associated with DDs. PMID- 14749616 TI - Update: influenza activity--United States, January 18-24, 2004. AB - The number of states reporting widespread influenza activity continued to decrease during the reporting week of January 18-24, 2004. One state health department reported widespread activity. A total of 20 states reported regional activity, 19 states and New York City reported local activity, and sporadic activity was reported by nine states, the District of Columbia, Guam, and Puerto Rico. The percentage of outpatient visits for influenza-like illness (ILI) remained the same during the week ending January 24. The percentage of specimens testing positive for influenza and the percentage of deaths attributed to pneumonia and influenza (P&I) decreased. PMID- 14749615 TI - Day care-related outbreaks of rhamnose-negative Shigella sonnei--six states, June 2001-March 2003. AB - During June 2001--March 2003, outbreaks of Shigella sonnei infections were reported in Delaware, Maryland, New Jersey, North Carolina, South Carolina, and Virginia. Five- to fortyfold increases in statewide shigellosis rates were observed during this period. These increases were attributed primarily to outbreaks in multiple day care settings that became prolonged and community wide. S. sonnei isolates from these states, as well as from New York City and Philadelphia, were similar genetically by pulsed-field gel electrophoresis (PFGE). Many of these isolates lacked the capacity to ferment rhamnose, which is unusual for S. sonnei. This report summarizes these outbreaks and describes the laboratory characteristics that link them. The findings underscore the importance of rapid and coordinated public health responses to isolated outbreaks of shigellosis. PMID- 14749618 TI - Genetics and prevention of pancreatic cancer. AB - BACKGROUND: Pancreatic cancer is an aggressive disease with a poor prognosis. Hereditary factors have been reported in up to 10% of cases of pancreatic cancer. The clinical characteristics and genetic abnormalities have been identified for a proportion of this high-risk group, and the development of preventive strategies for these individuals is now a primary goal of cancer clinicians. METHODS: A review of the current literature regarding the genetics, screening, and prevention of pancreatic cancer and its precursor lesions was undertaken. RESULTS: Risk factors for pancreatic cancer include smoking, chronic pancreatitis, and a genetic predisposition. The role of diabetes or a diet high in fat or meat remains unclear. The genetic mutations that accompany pancreatic cancer appear to occur in a temporal sequence, beginning in the earliest of precursor lesions. These mutations are detectable in pancreatic juice and, in conjunction with imaging, form the basis of screening programs for high-risk individuals. Not all precursor lesions will undergo malignant transformation, and testing is currently limited in its ability to determine which lesions will undergo transformation. CONCLUSIONS: Avoiding tobacco smoking and minimizing risk factors associated with chronic pancreatitis are recommended to reduce the risk of pancreatic cancer. Individuals with a high-risk genetic background require counseling, genetic testing if appropriate (BRCA2 mutation or p16INK4A inactivity) and secondary screening for pancreatic cancer in specialist centers. Risk stratification will improve as more genetic abnormalities causing pancreatic cancer are defined. PMID- 14749620 TI - Surgical management of early-stage pancreatic cancer. AB - BACKGROUND: Pancreatic cancer remains a difficult disease to treat. Diagnosis at an early stage may allow curative treatment with resection. In the past, the mortality associated with surgical treatment of pancreatic carcinoma was prohibitive but mortality associated with resection is now commensurate with all other major oncologic resections. Thus, the focus of surgical management has shifted to address several issues: the diagnosis and evaluation of patients with suspected pancreatic cancer, the role of preoperative endobiliary stenting, the role of laparoscopy, the extent of resection, the role of adjuvant and neoadjuvant treatment, and the role of specialized centers in treating the disease. METHODS: The current literature is reviewed to address these issues and help guide physicians who first encounter patients with suspected pancreatic cancer as well as surgeons who ultimately resect them. Practical evidence-based information to guide the decision-making process is provided. RESULTS: Surgical morbidity and mortality have achieved parity with other types of major oncologic resection, and a distinct survival advantage is possible when such therapy is applied early in the disease stage. Issues regarding the use of stents, extent of resection, and pre- vs post-operative chemoradiation therapy are becoming clearer as our collective experience broadens. CONCLUSIONS: Surgical treatment of pancreatic cancer should be aggressively pursued given the clearly established survival advantage and relief of symptoms achieved when it is applied appropriately. PMID- 14749619 TI - Applications of endoscopic ultrasonography in pancreatic cancer. AB - BACKGROUND: Accurate staging of pancreatic cancer is essential for surgical planning and for identification of locally advanced and metastatic disease that is incurable by surgery. Advances in endoscopic sonography (EUS), computed tomography (CT), and positron emission tomography have improved the accuracy of staging and reduced the number of incomplete surgical resections. Tissue acquisition is necessary in nonsurgical cases when chemoradiotherapy is considered. The complex regional anatomy of the pancreas makes cytologic diagnosis of malignancy at this region difficult without exploratory surgery. Although CT-guided fine-needle aspiration (FNA) is used for this purpose, reports of an increased risk of peritoneal dissemination of cancer cells and a false negative rate of nearly 20% make this a poor choice. The ability to position the EUS-transducer in direct proximity to the pancreas by means of the stomach and duodenum, combined with the use of FNA, increases the specificity of EUS in detecting pancreatic malignancies. METHODS: The current literature regarding the accuracy of EUS with FNA in the evaluation of pancreatic cancer is reviewed. RESULTS: EUS accuracy ranges from 78% to 94% for tumor staging and from 64% to 82% for nodal staging. EUS also enables FNA of lesions that are too small to be identified by CT or MRI or too well encased by surrounding vascular structures to safely allow percutaneous biopsy. The accuracy for detecting invasion into the superior mesenteric artery and vein is lower than that for detecting portal or splenic vein invasion, especially for large tumors. EUS permits delivery of localized therapy such as celiac plexus neurolysis for pain control and direct intra-lesional injection of antitumor therapy. CONCLUSIONS: EUS in combination with FNA is a highly accurate method of preoperative staging of pancreatic cancer, especially those too small to be characterized by CT or MRI, and it has the ability to obtain cytological confirmation of pancreatic cancer. PMID- 14749621 TI - Molecular-targeted agents in pancreatic cancer. AB - BACKGROUND: Despite the acceptance of gemcitabine as the standard first-line agent for the treatment of advanced pancreatic cancer as well as the improved response rates seen with gemcitabine combinations, novel therapies are needed for this disease, which has one of the lowest survival rates. The growing understanding of the molecular basis of pancreatic cancer and the recent introduction of targeted therapeutic agents have initiated novel studies that have the potential to improve on existing treatments. METHODS: We review the rationale and the clinical studies of therapeutic agents that target some of the molecular abnormalities commonly found in pancreatic cancer. RESULTS: Matrix metalloproteinase inhibitors (MMPIs), farnesyltransferase inhibitors (FTIs), and tyrosine-kinase inhibitors and monoclonal antibodies against growth factors or their receptors are novel agents that have undergone phase II or III trials. Phase III studies of MMPIs, alone or in combination with gemcitabine, and phase III studies of FTIs have produced disappointing results. Other agents in earlier phases of clinical development remain promising. CONCLUSIONS: Despite the negative studies of MMPIs and FTIs, the results of phase II trials of other drugs are encouraging. Targeted agents may improve the prognosis of pancreatic cancer. PMID- 14749622 TI - Palliative care in pancreatic cancer. AB - BACKGROUND: Pancreatic cancer is a formidable health problem, representing the 10th most common malignancy in the United States and the 4th most common cause of all cancer deaths. The overall 5-year survival rate is 4%, making this disease a model tumor in which to address the specialized care issues of palliative medicine. METHODS: General considerations in both medical decision-making and symptom management are reviewed. Treatment of patients with locally unresectable, recurrent, or metastatic disease is individualized, based on considerations that include patient age, patient wishes, family influence, insurance constraints, and geographic practice variations. RESULTS: Success in managing progressive symptoms is needed to palliate patients with advanced pancreatic cancer. Common problems include biliary obstruction, depression, pain, intestinal obstruction, and fatigue. CONCLUSIONS: Relief of pain and suffering associated with critical illness is required in managing patients with cancer. Pancreatic cancer is a model illness that mandates this need. PMID- 14749623 TI - 5-fluorouracil-induced coronary vasospasm. AB - BACKGROUND: Cardiotoxicity is a rare but well-documented adverse effect of 5 fluorouracil (5-FU). The underlying cause of this side effect of 5-FU is uncertain. METHODS: We present a case report of a 63-year-old man treated for metastatic colon cancer who experienced chest pain while being treated with the FOLFIRI regimen. This case report documents coronary artery spasm on catheterization observed with the continuous infusion of 5-FU. RESULTS: Cardiac catheterization obtained within 36 hours of the onset of chest pain revealed marked coronary vasospasm in the obtuse marginal coronary artery and a right coronary artery with a critical obstructive atherosclerotic plaque. Electrocardiogram revealed the myocardium area associated with the event was diffuse rather than localized to the right coronary artery. CONCLUSIONS: This observation supports the vasospastic hypothesis for 5-FU-induced angina. Although rare, this type of cardiotoxicity with 5-FU is a potentially lethal side effect. Therapy with 5-FU should be discontinued and patients should be promptly treated. PMID- 14749624 TI - The role of cancer cooperative groups within the spectrum of cancer care. PMID- 14749625 TI - Immunotherapy of surgical malignancies. PMID- 14749627 TI - Cannabinoids and the human uterus during pregnancy. AB - OBJECTIVE: The aim of this study was to investigate the expression of cannabinoid receptors in human uterine smooth muscle during pregnancy and to evaluate the effects of endogenous and exogenous cannabinoids on myometrial contractility in vitro. STUDY DESIGN: Human myometrial biopsy specimens were obtained at elective cesarean delivery and snap frozen or mounted for isometric recording under physiologic conditions. Cumulative doses of the endogenous cannabinoid anandamide or the exogenous cannabinoid Delta(9) (indicates a double bond between carbons 9 and 10) tetrahydrocannabinol were added in the range 1 nmol/L to 100 micromol/L. Selectivity of the cannabinoid receptor agonists was investigated with specific antagonists for the CB(1) and the CB(2) receptors. Reverse transcription polymerase chain reaction with primers for the CB(1) and CB(2) receptors was performed on messenger RNA that was isolated from human pregnant myometrium. RESULTS: Both anandamide and Delta(9)-tetrahydrocannabinol exerted a direct relaxant effect on human pregnant myometrium in vitro, which was of equal potency for both compounds. This relaxant effect was antagonized by the specific CB(1) receptor antagonist, SR 141716, but not by the specific CB(2) receptor antagonist, SR 144528 (n=6 specimens, P<.01). Both the CB(1) and CB(2) receptors are expressed in human myometrium. CONCLUSIONS: Both endogenous and exogenous cannabinoids exert a potent and direct relaxant effect on human pregnant myometrium, which is mediated through the CB(1) receptor. This highlights a possible role for endogenous cannabinoids during human parturition and pregnancy. These results also support the view that the use of exogenous cannabinoids during pregnancy is not linked independently with preterm labor. PMID- 14749628 TI - Unnecessary cesarean delivery in Louisiana: an analysis of birth certificate data. AB - OBJECTIVE: The purpose of this study was to explore the temporal trends and factors that are associated with cesarean deliveries and potentially unnecessary cesarean deliveries. STUDY DESIGN: The Louisiana birth certificate database was evaluated to identify a total of 57 potential indications/risk factors and maternal demographic factors that are associated with methods of delivery over the period from January 1993 to December 2000. A cesarean delivery without any potential indications/risk factors in the birth certificate was classified as unnecessary. RESULTS: The primary cesarean delivery rate decreased and the repeat cesarean delivery rate increased significantly during the study period. But neither the absence nor the presence of potential indications/risk factors accounted for these changes. The average potentially unnecessary primary and repeat cesarean deliveries in Louisiana were 17 and 43, respectively, per 100 cesarean deliveries over the years 1993 through 2000. CONCLUSION: The proportions of potentially unnecessary cesarean deliveries are relatively high in Louisiana. It is important to explore the influence of nonclinical factors on unnecessary cesarean delivery to reduce the cesarean rates. PMID- 14749629 TI - Abdominal sacral colpopexy or vaginal sacrospinous colpopexy for vaginal vault prolapse: a prospective randomized study. AB - OBJECTIVE: The purpose of this study was to compare the abdominal sacral colpopexy and vaginal sacrospinous colpopexy in the treatment of vaginal vault prolapse. STUDY DESIGN: Ninety-five women with vaginal vault prolapse were allocated randomly to sacral colpopexy (47 women) or sacrospinous colpopexy (48 women). Primary outcome measurements include subjective, objective, and patient determined success rates. Secondary outcomes include the impact on bowel, bladder, and sexual function, cost, and quality of life. RESULTS: Two years after the operation (range, 6-60 months), the subjective success rate was 94% in the abdominal and 91% in the vaginal group (P=.19). The objective success rate was 76% in the abdominal group and 69% in the vaginal group (P=.48). The abdominal approach was associated with a longer operating time, a slower return to activities of daily living, and a greater cost than the sacrospinous colpopexy (P<.01). Both surgeries significantly improved the patient's quality of life (P<.05). CONCLUSION: Abdominal sacral colpopexy and vaginal sacrospinous colpopexy are both highly effective in the treatment of vaginal vault prolapse. PMID- 14749630 TI - Progression and remission of pelvic organ prolapse: a longitudinal study of menopausal women. AB - OBJECTIVE: The purpose of this study was to describe the natural history of pelvic organ prolapse after menopause. STUDY DESIGN: Over 2 to 8 years, participants in the estrogen plus progestin trial of the Women's Health Initiative at the University of California Davis had annual pelvic examinations, with an assessment of uterine prolapse, cystocele, and rectocele. The findings from these examinations were used to describe the incidence of pelvic organ prolapse, the probability of progression or regression, and the associated risk factors. RESULTS: At baseline, 31.8% of women had pelvic organ prolapse (n=412 women). The annual incidences of cystocele, rectocele, and uterine prolapse were 9.3, 5.7, and 1.5 cases per 100 women-years, respectively. Incident prolapse was associated with increasing parity and waist circumference. The progression rates for grade 1 pelvic organ prolapse (per 100 women-years) were 9.5 for cystocele, 13.5 for rectocele, and 1.9 for uterine prolapse. The annual rates of regression (per 100 women-years) was 23.5, 22, and 48, respectively. CONCLUSION: Our data suggest that pelvic organ prolapse is not always chronic and progressive as traditionally thought. Spontaneous regression is common, especially for grade 1 prolapse. PMID- 14749631 TI - Ambulatory procedures for urinary incontinence in the United States, 1994-1996. AB - OBJECTIVE: The objective of our study was to describe the national rates of ambulatory surgery for urinary incontinence in women. STUDY DESIGN: We used the National Survey of Ambulatory Surgery, a federal database that samples outpatient surgery in the United States. Data from 1994 to 1996 were analyzed for diagnoses and procedures coded using the International Classification of Diseases, 9th revision, Clinical Modification, classification system. Age-adjusted rates were calculated with use of the 1990 census population and compared for change over time by using the score test for linear trend. RESULTS: The estimated number of women undergoing outpatient surgery for urinary incontinence increased from 7,200 in 1994 to 15,900 in 1996 (P=.3), whereas the age-adjusted rate remained stable at approximately 10 procedures per 100,000 women per year. The procedures performed included "other repair of the bladder" (ICD-9-CM code 57.89), "other repair of urinary stress incontinence" (code 59.7), plication of the urethrovesical junction, suprapubic sling operation, retropubic urethral suspension, paraurethral suspension, and levator muscle operation. The mean age of women undergoing these procedures was 58+/-14.5 years. Women undergoing incontinence procedures were 76% white, 0.3% African American, and 24% other (including unknown). Ninety-two percent of the procedures were completed at hospitals, whereas 8% were performed at free-standing ambulatory surgery centers. CONCLUSION: Between 1994 and 1996, the number of ambulatory surgeries for urinary incontinence in women doubled. PMID- 14749632 TI - Laparoscopic occlusion of uterine vessels for the treatment of symptomatic fibroids: Initial experience and comparison to uterine artery embolization. AB - OBJECTIVE: Our purpose was to evaluate the effects of laparoscopic occlusion of uterine vessels in treating symptomatic fibroids and compare with embolization of the uterine arteries. STUDY DESIGN: We studied 46 premenopausal women, aged 43 (34-51) years with symptomatic uterine fibroids, undergoing radiologic embolization (n=24) and laparoscopy closure of the uterine arteries (n=22). RESULTS: The laparoscopic technique reduced picture blood assessment score after 6 months by 50% from an initial value of 345 (+/-288). Uterus volume was reduced by 37% (+/-18%), and the dominant fibroid was reduced by 36% (+/-31%). Postoperative pain and use of pain relief differed significantly, requiring more pain medication after embolization: ketobemidon 38 mg compared with 16 mg in the laparoscopic group (P=.008). Specific complications to the laparoscopic technique were temporary damage to the obturator nerve in three patients. CONCLUSION: Laparoscopic occlusion of uterine vessels is a promising new method for treating fibroid-related symptoms, with less postoperative pain than embolization and comparable effects on symptoms. PMID- 14749633 TI - In vitro effect of gonadotropin-releasing hormone agonist on natural killer cell cytolysis in women with and without endometriosis. AB - OBJECTIVE: The purpose of this study was to assess the in vitro effect of gonadotropin-releasing hormone agonist on natural killer cell activity in women with and without endometriosis and to ascertain whether gonadotropin-releasing hormone agonist effects on natural killer cell activity are direct or mediated solely through the hypoestrogenic state that they produce in vivo. STUDY DESIGN: With use of a chromium 51 release microcytotoxicity assay with K562 target cells, natural killer cell activity was measured after the incubation of mononuclear cells with leuprolide acetate that was obtained from 16 patients with endometriosis and 11 control subjects. RESULTS: The cytotoxicity of natural killer cells that were obtained from patients with endometriosis was reduced significantly (P<.001) with leuprolide. Natural killer cell cytotoxicity from control patients was also significantly decreased (P=.005) with gonadotropin releasing hormone agonist. Natural killer cell cytotoxicity was significantly lower in patients with endometriosis than in control patients (P=.029). CONCLUSION: These findings suggest a direct immunomodulatory role of gonadotropin releasing hormone agonist on natural killer cell activity and confirm previous findings that patients with endometriosis have reduced natural killer cell cytotoxicity. PMID- 14749634 TI - Pregnancy during the use of levonorgestrel intrauterine system. AB - OBJECTIVES: This study was undertaken to evaluate the pregnancy rate with the levonorgestrel intrauterine system (LNG IUS) and to analyze the outcome of pregnancies with the LNG IUS in situ in regular use. STUDY DESIGN: Questionnaires from 17,360 users of the LNG IUS were analyzed. With the consent of women reporting pregnancy during the use of the LNG IUS, related hospital records were reviewed concerning the pregnancies. RESULTS: Originally 132 pregnancies were reported. Medical records were reviewed from 108 of these women. In 64 pregnancies, conception occurred with the LNG IUS in situ. Thirty-three pregnancies were ectopic. The 5-year cumulative pregnancy rate per 100 users was 0.5 and the 5-year Pearl rate was 0.11. CONCLUSION: Pregnancy with the LNG IUS in situ is rare. Ectopic pregnancies constitute 53% of all pregnancies. Typical pregnancy symptoms occur during pregnancies with the LNG IUS. The importance to counsel about the risk of pregnancy before insertion is emphasized. PMID- 14749635 TI - A randomized controlled comparison of sublingual and vaginal administration of misoprostol for cervical priming before first-trimester surgical abortion. AB - OBJECTIVE: The purpose of this study was to compare the effectiveness of the sublingual and vaginal administration of misoprostol for cervical priming before surgical termination of pregnancy. STUDY DESIGN: In a randomized controlled trial, 74 primigravid women who were undergoing surgical abortion were assigned randomly to receive misoprostol (400 microg) sublingually or vaginally. RESULTS: There was no statistically significant difference in the cumulative force that was required to dilate the cervix to 9 mm, for baseline cervical dilatation, for priming to abortion interval, for operating time, or for intraoperative blood loss between the two groups. Women in the sublingual group had more nausea (P=.008), vomiting (P=.01), diarrhea (P=.01), and unpleasant mouth taste (P=.0001) compared with the women in the vaginal group. In the sublingual group, 65% of women were satisfied with the route of misoprostol administration compared with 78% in the vaginal group (P=.11). Most of the staff members (84%) said that they would recommend the sublingual administration of misoprostol (P=.0001). CONCLUSION: The sublingual administration of misoprostol is an effective alternative to vaginal administration for cervical priming before surgical abortion; despite a higher incidence of side effects, there was high patient and staff acceptability. PMID- 14749636 TI - The novel serine protease tumor-associated differentially expressed gene-14 (KLK8/Neuropsin/Ovasin) is highly overexpressed in cervical cancer. AB - OBJECTIVE: Serine proteases are redundant enzymes implicated in the extracellular modulation required for tumor growth and invasion. Tumor-associated differentially expressed gene-14 (TADG-14) is a novel transmembrane serine protease recently reported by our group to be highly overexpressed in ovarian carcinomas. The goal of this study was to investigate the frequency of expression of the TADG-14 gene in human cervical tumors. STUDY DESIGN: TADG-14 expression was evaluated in 19 cervical cancer cell lines (11 primary and 8 established cell lines) as well as in 8 normal cervical keratinocyte cultures by reverse transcriptase polymerase chain reaction. In addition, to validate gene expression data at the protein level, TADG-14 expression was evaluated by immunohistochemistry on paraffin-embedded tissue from which all 11 primary tumor cell lines were established. RESULTS: TADG-14 was found to be highly expressed in 82% (9/11) primary cervical cancer cell lines and in 87% (7/8) established cervical cancer cell lines by reverse transcriptase-polymerase chain reaction. Expression of TADG-14 by primary squamous cervical tumors was 100% (6/6), whereas 60% (3/5) of primary adenocarcinomas expressed TADG-14. In contrast, none of the normal cervical keratinocyte control cultures (n=4) or flash frozen normal cervical biopsy specimens (n=4) expressed TADG-14. Immunohistochemistry staining of paraffin-embedded cervical cancer specimens confirmed TADG-14 expression in tumor cells and its absence on normal cervical epithelial cells. CONCLUSION: Cervical cancer expressed a high level of TADG-14, suggesting that this protease may play an important role in invasion and metastasis. Because TADG-14 appears only in abundance in tumor tissue and contains a secretion signal sequence, suggesting that TADG-14 is secreted, it may prove to be a useful diagnostic tool for the early detection of recurrent/persistent cervical cancer after standard treatment or as a novel molecular target for cervical cancer therapy. PMID- 14749637 TI - Glassy cell adenocarcinoma of the uterine cervix. AB - OBJECTIVE: Glassy cell carcinoma of the cervix is a rare variant of cervical cancer accounting for a small percentage of cell types. This study was undertaken to review our experience with glassy cell adenocarcinoma. STUDY DESIGN: Review of adenocarcinomas from the period 1970 to 1985 was undertaken. Pathology review and survival analysis was performed comparing the glassy cell adenocarcinomas with other adenocarcinoma cell types. RESULTS: Twenty-one patients with glassy cell adenocarcinoma were identified. The mean age was 44 years. The stage distribution showed stage I=10, stage II=8, stage III=1, and stage IV=2. The cumulative survival according to stage was stage I=50%, stage II=36%, stage III=0%, and stage IV=0%. A separate analysis was performed for stage I comparing glassy cell carcinoma with other stage I adenocarcinoma cell types. The survival for stage I nonglassy cell adenocarcinoma was 61% compared with glassy cell adenocarcinoma 48% (n=10) (P=.5). A separate subset for adenosquamous carcinoma alone was performed. The survival for stage I adenosquamous carcinoma (n=30) was 58%. The survival for adenosquamous glassy cell (n=5) was 30% (P=.45). CONCLUSION: Glassy cell adenocarcinoma is a small subset of adenocarcinoma. When compared with other adenocarcinomas, survival is not significantly decreased. PMID- 14749638 TI - Physical injury after sexual assault: findings of a large case series. AB - OBJECTIVE: This study was undertaken to determine characteristics associated with physical injury in female sexual assault victims. STUDY DESIGN: All females who were 15 years or older presenting after sexual assault to an urban emergency department during a 34-month period underwent standardized evaluation. Analysis was performed by chi(2) and logistic regression. RESULTS: Of 819 women, 52% had general body and 20% had genital-anal trauma; 41% were without injury. General body trauma was independently associated with being hit or kicked (odds ratio [OR]=7.7, 95% CI, 5.1-11.7), attempted strangulation (OR=4.2, 95% CI, 2.5-7.2), oral or anal penetration (OR=1.7, 95% CI, 1.2-2.3), and stranger (OR=2.4, 95% CI, 1.7-3.4) assault. Genital-anal injury was more frequent in victims younger than 20 and older than 49 years (P<.05), in virgins (OR=2.7, 95% CI, 1.4-5.4) and those examined within 24 hours (OR=1.7, 95% CI, 1.2-2.4) and after anal assault (OR=1.7, 95% CI, 1.1-2.6). CONCLUSION: General body injury is primarily associated with situational factors, whereas genital-anal injury is less frequent and related to victim age, virginal status, and time to examination. PMID- 14749639 TI - Effects of estrogen therapy on hearing in postmenopausal women. AB - OBJECTIVE: This study was undertaken to investigate how hormone therapy affects hearing in postmenopausal women. STUDY DESIGN: This prospective study involved 109 postmenopausal women. Twenty of the women were using estrogen therapy (ET group), 30 women were using hormone therapy (HT group), and 59 had not received hormone therapy of any kind (control group). Otoscopic examination revealed normal tympanic membranes in all 109 subjects. Each individual was tested with low- (250-2000 Hz) and high-frequency audiometry (4000-16000 Hz). Duration of hormone therapy was recorded, and patient characteristics (age, type of menopause, time since onset of menopause), body mass index (BMI), and hearing test results in the ET, HT, and control groups were compared. RESULTS: There were no statistically significant differences between the treatment (ET and HT group) and control groups with respect to age, BMI, or time since onset of menopause. The mean time on HT and ET was 4.13+/-2.41 years and 3.35+/-2.20 years, respectively. The mean air conduction results at low frequencies (250, 500, 1000, and 2000 Hz) in the ET group were significantly higher than the corresponding findings in the control group (P<.001) and than the HT group (P<.001). When the same comparisons were made between the HT group and the control group, none of the differences was statistically significant (P>.05). The mean air-conduction results at high frequencies (4, 6, 8, 10, 12, 14, and 16 kHz) in the ET group were significantly higher than the corresponding results in the HT group (P<.008). ET versus controls and HT versus controls at high frequencies revealed no significant differences (P>.05). The mean bone conduction results in the ET group were significantly higher than the corresponding findings in the control group (P<.016). Analysis of the same comparisons between the HT-ET and HT-control groups revealed no significant differences (P>.05). CONCLUSION: Estrogen therapy may slow down hearing loss in aging postmenopausal women; however, further studies of larger series are needed to confirm this, and the sites of hormonal action must also be explored. PMID- 14749640 TI - Knot integrity with nonidentical and parallel sliding knots. AB - OBJECTIVE: The study was performed to test the knot integrity of parallel and nonidentical sliding knots with the use of monofilament suture. STUDY DESIGN: We investigated the proportion of parallel sliding knots that become untied compared with nonidentical sliding knots in four different monofilament suture configurations: group A, 6 throws, 0-0 gauge; group B, 10 throws, 0-0 gauge; group C, 6 throws, 2-0 gauge; group D, 10 throws, 2-0 gauge. Each combination was tested 20 times, for a total of 160 knots. RESULTS: The nonidentical sliding knot untied 40% of the time in group A compared with 0% for the parallel knot (P=.003). In the other 6-throw group, group C, 45% of the nonidentical knots became untied compared with 0% of the parallel knots (P=.001). There were no significant differences in the proportions of untying for each knot type in the 10-throw groups. CONCLUSION: Parallel sliding knots demonstrate superior knot integrity compared with nonidentical sliding knots with monofilament suture. The addition of throws to the nonidentical knot improves its knot security. PMID- 14749641 TI - In vitro lesion repair by human amnion epithelial and mesenchymal cells. AB - OBJECTIVE: The purpose of this study was to compare wound healing by human amnion epithelial and mesenchymal cells from preterm and term placenta with the use of an in vitro lesion repair assay. STUDY DESIGN: Lesions were created in confluent monolayers of amnion epithelial and mesenchymal cells from preterm and term placentas. The repair was monitored by the measurement of the lesion area and the response to potential stimulants (platelet-derived growth factor, tumor necrosis factor-alpha, fibrinogen, and phorbol myristate acetate). Cell proliferation was detected with 5-bromodeoxyuridine staining. RESULTS: Lesion repair was complete within 40 hours in control epithelial cultures from preterm and term placenta but incomplete in mesenchymal cultures (preterm cells, 80%; term cells, 40%). Platelet-derived growth factor, tumor necrosis factor-alpha, fibrinogen, and phorbol myristate acetate did not accelerate repair in either cell type. CONCLUSION: An in vitro lesion repair assay revealed the differences in lesion repair capacity between amnion epithelial and mesenchymal cells and between mesenchymal cells from preterm and term placenta. PMID- 14749642 TI - Clinical signs predict 30-month neurodevelopmental outcome after neonatal encephalopathy. AB - OBJECTIVE: This study was undertaken to determine the value of a neonatal encephalopathy score (ES) and the presence of seizures for predicting 30-month neurodevelopmental outcome. STUDY DESIGN: In a cohort study, 68 term newborn infants with encephalopathy were evaluated with an ES based on alertness, feeding, tone, respiratory status, reflexes, and seizure activity (range: 0-6). Seizures were noted as present or absent clinically. Significant cognitive deficits (Mental Development Index <70), motor disability (spastic triplegia/quadriplegia), or death were abnormal outcomes. RESULTS: Twenty-two newborn infants (32%) had abnormal outcomes. With the use of maximum ES and presence of seizures from days 1 to 3 of life, 87% of newborn infants were correctly classified (area under receiver operating curve 0.93). By using ES and presence of seizures on day 1 only, 87% of newborn infants were correctly classified (area under receiver operating curve 0.89). CONCLUSION: The severity of neonatal encephalopathy and the presence of seizures are valuable predictors of 30-month neurodevelopmental outcome, as early as the first day of life. PMID- 14749643 TI - Accuracy of home pregnancy tests at the time of missed menses. AB - OBJECTIVE: The purpose of this study was to investigate the validity of home pregnancy tests (HPTs) around the time of the missed menses. STUDY DESIGN: Levels of human chorionic gonadotropin (hCG) were determined in urine around the time of the missed menses, and 5th and 95th centiles were calculated. In a blind study evaluating 18 brands of HPTs, each was tested six times with five concentrations of hCG, 0 to 100 mIU/mL. Sensitivity was defined as the concentration at which six of six brands gave positive results, at the suggested reading time or at an extended time. RESULTS: It was estimated that a sensitivity of 12.5 mIU/mL was needed to detect 95% of pregnancies at the time of missing menses. Only 1 of 18 HPTs had this sensitivity. If faintly discernible results considered, 2 of 18 brands gave false-positive or invalid results. Clear positive results were given by only 44% of the brands at 100 mIU/mL hCG. If one accepted faintly discernible and extended reading times, all brands responded at 100 mIU/mL. A test with 100 mIU/mL sensitivity would detect approximately 16% of pregnancies at the time of missed menses. CONCLUSION: The utility of HPTs is questioned. Clinicians need to be aware of the limitations of current HPT brands. PMID- 14749644 TI - Subcapsular liver hematoma in HELLP syndrome: Evaluation of diagnostic and therapeutic options--a unicenter study. AB - OBJECTIVE: Subcapsular liver hematoma formation has been reported in less than 2% of pregnancies complicated by HELLP (hemolysis, elevated liver enzymes, low platelets) syndrome. The purpose of this study was to identify the main diagnostic and therapeutic options for management of these patients. STUDY DESIGN: In this 10-year retrospective review, we performed a computer-directed search of all cases of confirmed HELLP syndrome with hepatic hematoma treated in the surgical department of our tertiary care referral medical center. RESULTS: Five patients with subcapsular liver hematoma in HELLP syndrome could be identified. All patients received transabdominal ultrasound, computed tomography, or magnetic resonance imaging. Conservative treatment was successful in three patients. Emergency surgical intervention was necessary in two patients, including one liver transplantation. CONCLUSION: The case series shows the full diagnostic spectrum with transabdominal ultrasound, computed tomography, and magnetic resonance imaging, as well as the different therapeutic options varying from conservative therapy to operative management, including liver transplantation. PMID- 14749645 TI - Dystocia in nulliparous patients monitored with fetal pulse oximetry. AB - OBJECTIVE: A critical analysis of the United States randomized controlled trial of fetal pulse oximetry concluded that nonreassuring fetal heart rate patterns used for study entry may have been a marker for dystocia. We prospectively studied nulliparous women in labor whose progress was monitored with fetal pulse oximetry to examine the relationship between nonreassuring fetal heart rate patterns and operative delivery for dystocia. STUDY DESIGN: A prospective nonrandomized observational cohort study compared two distinct classes of nonreassuring fetal heart rate patterns (class I: intermittent, mildly nonreassuring; class II: persistent, progressive, and moderate to severely nonreassuring) among nulliparous patients with the use of fetal pulse oximetry to confirm fetal well-being. Definitions of dystocia included the cessation of labor progress in the first (3 hours) or second (2 hours) stage of labor, despite adequate uterine activity that was assessed with an intrauterine pressure catheter. Independent review confirmed the classification of nonreassuring fetal heart rate patterns and study entry criteria. RESULTS: Two hundred seventy-four patients met study criteria and had sufficient information for fetal heart rate tracing interpretation. Two hundred thirty-seven patients (86.5%) were class II, and 37 patients (13.5%) were class I. The two classes of patients were comparable in a variety of obstetric, demographic, and perinatal variables. Twelve percent of all patients were delivered for nonreassuring fetal status. Significantly more class II patients (22%) were delivered by cesarean for dystocia than were class I patients (8%). Higher doses and a longer number of hours of oxytocin were required among class II patients. Significantly more occiput posterior positions were noted among all patients who underwent cesarean delivery for dystocia compared with other modes of delivery. CONCLUSION: Significantly nonreassuring fetal heart rate patterns predict cesarean delivery for dystocia among nulliparous patients with normally oxygenated fetuses in a setting of a standardized labor management protocol. This confirms the observations in the randomized controlled trial of fetal pulse oximetry in the United States and may provide insight into the treatment of nonprogressive labor in contemporary practice. PMID- 14749646 TI - Unselected low-risk pregnancies and the effect of continuous intrapartum fetal heart rate monitoring on umbilical blood gases and cerebral palsy. AB - OBJECTIVE: Our purpose was to evaluate the clinical validity of electronic fetal heart rate monitoring to detect fetal acidemia and to evaluate the prevalence of cerebral palsy in unselected low-risk pregnancies. STUDY DESIGN: We selected two secondary and two tertiary level institutions in which 10,030 infants were born. Among them, 5546 of the pregnancies were low-risk pregnancies by antepartum evaluation. The fetal heart rate patterns were interpreted according to the guidelines of the National Institute of Child Health and Human Development. The correlations between the fetal heart rate pattern and umbilical blood gases and the fetal heart rate pattern and cerebral palsy were studied. Spastic cerebral palsy was diagnosed at > or =1 year by pediatric neurologists. Statistics included unpaired t test, contingency table with chi(2) and Fisher tests, and one way analysis of variance with Bonferroni/Dunn test. RESULTS: On the basis of the severity of decelerations, frequency of decelerations, and decreased variability, umbilical pH, and Po(2) level were decreased accordingly, and incidence of pH<7.1 was increased. Sensitivity and false-positive rate of nonreassuring fetal heart rate patterns for fetal acidemia were 63% and 89%. There were nine cerebral palsy cases: six of the cases were preexisting asphyxia before monitoring was initiated, two of the cases were cytomegaloviral infections, and one of the cases was a maternal amniotic fluid embolism. CONCLUSION: In low-risk pregnancies, intrapartum fetal heart rate monitoring was useful to detect fetal acidemia. Cerebral palsy caused by intrapartum asphyxia was restricted to unavoidable accidents under continuous fetal heart rate monitoring. PMID- 14749647 TI - Randomized trial between two active labor management protocols in the presence of an unfavorable cervix. AB - OBJECTIVE: The purpose of this study was to compare the efficacy of two protocols for active management of labor at term in the presence of an unfavorable cervix. STUDY DESIGN: Pregnancies that underwent labor induction at > or =37 weeks of gestation with an unfavorable cervix (Bishop score, < or =6) were randomly assigned to receive vaginally either a single dose of sustained-release dinoprostone (Cervidil) with concurrent low-dose oxytocin or multidosing of misoprostol (25 microg every 4 hours) followed by high-dose oxytocin. The primary outcome was the time interval from induction to vaginal delivery. Other parameters included excess uterine activity and cesarean delivery rates. RESULTS: A total of 151 patients (dinoprostone, 74 patients; misoprostol, 77 patients) were enrolled. The mean time from the initiation of induction to vaginal delivery was the same in the dinoprostone and misoprostol groups (15.7 hours; 95% CI, 13.7 17.7 hours vs 16.0 hours; 95% CI, 14.1-17.8 hours; P=.34), regardless of parity. The dinoprostone and misoprostol groups did not differ statistically in the percent of patients who were delivered vaginally by 12 hours (36.2% vs 29.7%), 18 hours (63.8% vs 56.3%), and 24 hours (81.0% vs 81.3%). Excess uterine activity was not more common in either group, and hyperstimulation syndrome was absent in all cases. Primary cesarean delivery rates were similar (dinoprostone, 21.6%; misoprostol, 16.9%; relative risk, 1.3; 95% CI, 0.7-2.5), with a failed induction that occurred in one case in each group. CONCLUSION: Sustained-release dinoprostone with concurrent low-dose oxytocin and intermittent misoprostol with delayed high-dose oxytocin are effective alternatives for active management of labor with an unfavorable cervix. PMID- 14749648 TI - Risk of cesarean delivery in nulliparous women at greater than 41 weeks' gestational age with an unengaged vertex. AB - OBJECTIVE: The purpose of this study was to determine whether an unengaged vertex significantly increased the risk of cesarean delivery in nulliparous patients at 41 weeks or greater. STUDY DESIGN: The medical records from all nulliparous patients greater than 41 weeks' gestation delivered at a single institution were reviewed. Patients undergoing both spontaneous and induced labor were included. Multivariate analyses were used to compare the influence of admission fetal station versus induction of labor on the risk of cesarean delivery. RESULTS: Four hundred forty-eight nulliparous women at greater than 41 weeks' gestation were delivered at our institution during the study period. Sixty-two percent of these patients underwent induction of labor. There was a statistically significant increase in cesarean delivery rate compared with station (6% of patients at -1 station, 20% at -2 station, 43% at -3 station, and 77% at -4 station; P=.001). Compared with patients with an engaged vertex, patients with an unengaged vertex had 12.4 times the risk of cesarean delivery. Most of the cesarean deliveries were performed for failure to progress. On the basis of multivariate analysis, the odds of cesarean delivery were better predicted by fetal station than induction of labor. CONCLUSION: Nulliparous patients at 41 weeks or greater with an unengaged vertex are 12.4 times more likely to be delivered by cesarean section than a patient with an engaged vertex. PMID- 14749649 TI - Omphalocele: clinical outcomes in cases with normal karyotypes. AB - OBJECTIVE: This study was undertaken to examine neonatal outcomes in karyotypically normal cases of omphalocele with respect to mode of delivery, presence of associated anomalies, presence of extracorporeal liver, and method of abdominal wall closure. STUDY DESIGN: We reviewed 36 cases of omphalocele with delivery at University of North Carolina Hospitals between 1988 and 2001. Elective terminations and pregnancies resulting in miscarriage before 20 weeks' gestation were excluded. RESULTS: Perinatal mortality rate was 19%. Rate of composite neonatal morbidity was 25%. Associated major anomalies were present in 11 (31%) cases and were associated with increased neonatal mortality. Cesarean deliveries were performed in 21 (58%) cases and were not associated with increased primary closure rates or decreased neonatal morbidity and mortality. Extracorporeal liver was present in 27 (75%) cases and was associated with decreased rates of primary closure but did not affect neonatal outcome. CONCLUSION: In cases of ongoing omphalocele, perinatal mortality rates are low in the absence of associated anomalies or genetic defects. Intracorporeal liver was not associated with increased rates of associated anomalies or was it associated with increased neonatal morbidity or mortality. PMID- 14749650 TI - Vaginal fetal fibronectin as a predictor of spontaneous preterm delivery after multifetal pregnancy reduction. AB - OBJECTIVE: The study was undertaken to assess the validity of vaginal fetal fibronectin assay as a screening test for spontaneous preterm delivery in asymptomatic patients who have undergone multifetal pregnancy reduction (MFPR). STUDY DESIGN: A historic cohort of 63 patients who underwent MFPR between 10 and 14 weeks of gestation was identified. All patients underwent serial vaginal fetal fibronectin sampling every 2 to 3 weeks from 22 weeks of gestation until delivery or 32 weeks of gestation. The fetal fibronectin concentration was measured by enzyme-linked immunosorbent assay, with 50 ng/mL or greater indicating a positive result. Charts were reviewed for fetal fibronectin results and pregnancy outcome data. Groups were compared by use of Fisher exact test. RESULTS: There were 13 singleton and 50 twin gestations after MFPR. A median of 4 fetal fibronectin assays were performed per patient. A total of 234 fetal fibronectin assays were performed with 222 (94.9%) negative results and 12 (5.1%) positive results. Overall, 41.3% of gestations were delivered spontaneously before 37 weeks; 7.9% were delivered before 34 weeks. The mean interval between tests was 17.8 days (+/ 7.2 days). For delivery within 2 and 3 weeks of a single test, fetal fibronectin had a sensitivity of 66.7% and 50%, a specificity of 95.7% and 96.1%, a positive predictive value of 16.7% and 25%, and a negative predictive value of 99.5% and 98.6%, respectively. CONCLUSION: The fetal fibronectin test has similar validity to predict spontaneous preterm delivery in these high-risk pregnancies as in previously published cohorts. PMID- 14749651 TI - A fetal response to chorioamnionitis is associated with early survival after preterm birth. AB - OBJECTIVE: The purpose of this study was to determine, in a large preterm cohort (20-34 completed weeks of gestation), the incidence of histologic chorioamnionitis and the incidence of a histologic fetal response to chorioamnionitis (umbilical vasculitis with or without funisitis) in neonatal survivors (to 28 days) and perinatal deaths. STUDY DESIGN: Placental histopathology was reviewed (n=3928 reports). In a subset of this cohort (n=2076 reports), evidence of a histologic fetal response was compared in neonatal survivors and perinatal deaths. RESULTS: The incidence of histologic chorioamnionitis ranged from 66% at 20 to 24 weeks of gestation (n=261 neonates) to 16% at 34 weeks (n=770 neonates). The overall incidence was 31% (n=3928 neonates). At 25 to 29 weeks of gestation, neonatal survivors had a higher incidence of histologic chorioamnionitis (P=.02; 95% CI, 1.02-1.21). In addition, neonatal survivors had a higher incidence of a histologic fetal response to chorioamnionitis at 25 to 29 weeks of gestation (P=.01; 95%CI, 0.33-0.86) and 30 to 34 weeks of gestation (P=.02; 95%CI, 0.18-0.85). CONCLUSION: Histologic chorioamnionitis is inversely related to gestational age. Both histologic chorioamnionitis and a histologic fetal response to chorioamnionitis were observed to be more common in preterm survivors of the neonatal period. PMID- 14749652 TI - Intraobserver and interobserver variability for the histologic diagnosis of chorioamnionitis. AB - OBJECTIVES: The interobserver and intraobserver variability for the histopathologic diagnosis of chorioamnionitis was examined. STUDY DESIGN: Two examiners independently reviewed archived slides from 250 placentas. They were blinded to the original diagnosis and details of the pregnancy. Definitions for two important diagnoses were made and a protocol for recording information used in trial before the study. Slides were examined under conditions reflecting normal working practice. RESULTS: A high level of agreement for the diagnosis of chorioamnionitis and umbilical vessel vasculitis (kappa range 0.78-0.81) between the examiners was found. Comparison of the agreement between each examiner and their original diagnosis was also excellent (kappa range 0.90-0.91). CONCLUSION: This study has audited the reliability of the diagnosis of chorioamnionitis, a common and important placental finding. The reproducible and reliable degree of agreement demonstrated permits further research to be undertaken relating this diagnosis to adverse postnatal outcomes. PMID- 14749653 TI - Neuromorbidity in preterm twins in relation to chorionicity and discordant birth weight. AB - OBJECTIVE: The purpose of this study was to determine the incidence of neurologic morbidity in preterm monochorionic (MC) and dichorionic (DC) twins. STUDY DESIGN: We collected perinatal, neonatal, and infant follow-up data of 76 MC and 78 DC twins born between 24 and 34 weeks of gestation (295 infants). Risks of neuromorbidity in the surviving infants were evaluated in relation to chorionicity, discordant birth weight (>20%), twin-twin transfusion syndrome (TTTS), and cotwin death. RESULTS: The overall incidence of cerebral palsy and minor neurologic disabilities in surviving twins was 4% and 9%, respectively. MC infants had a higher incidence of cerebral palsy (8% vs 1%, P<.05) and neurologic morbidity (15% vs 3%, P<.05) than DC infants. The risk of impaired neurodevelopment was higher in MC infants with discordant birth weight (42%, P<.01), TTTS (37%, P<.01), and cotwin death (60%, P<.01) than those with concordant birth weight (8%). In MC pregnancies, the cerebral palsy risk was higher in infants with discordant birth weight than those with chronic TTTS (19% vs 4%, P<.05). Similarly, discordant DC infants had higher neuromorbidity than concordant group (5% vs 1%, P<.05). In both MC and DC discordant infants, neurologic morbidity was independent of growth restriction. CONCLUSION: Neurologic morbidity in the preterm MC infants was 7-fold higher than DC infants because of chronic TTTS, discordant birth weight, and cotwin death in utero. PMID- 14749654 TI - How well do the amniotic fluid index and single deepest pocket indices (below the 3rd and 5th and above the 95th and 97th percentiles) predict oligohydramnios and hydramnios? AB - OBJECTIVE: This study was undertaken to determine whether the amniotic fluid index (AFI) and single deepest pocket percentiles can reliably identify a dye determined amniotic fluid volume. STUDY DESIGN: Ultrasound measurements and dye determined volumes were prospectively collected. Oligohydramnios predictors included AFI and single deepest pocket below the 3rd and 5th percentiles, and hydramnios predictors were based on AFI and single deepest pocket above the 95th and 97th percentiles. RESULTS: Two hundred ninety-one women with singleton pregnancies participated, including 75 pregnancies (26%) with oligohydramnios and 31 pregnancies (10%) with hydramnios. The predictive ability of the AFI and single deepest pocket indices (<3rd and 5th percentiles) to identify oligohydramnios was between 11% and 27% and to identify hydramnios (>95th and 97th percentiles) ranged between 33% and 46%. The best prediction was for normal fluid volume with an accuracy of 83% to 94%. CONCLUSION: Amniotic fluid indices perform best for the identifying normal AF volumes, whereas the identification of oligohydramnios or hydramnios is poor. PMID- 14749655 TI - Neonatal mortality rates among growth-discordant twins, classified according to the birth weight of the smaller twin. AB - OBJECTIVE: The purpose of this study was to evaluate neonatal mortality rates among discordant twins, classified according to the birth weight of the smaller twin. STUDY DESIGN: We compared neonatal mortality rates among three groups of discordant twins (>25%), distinguished by the birth weight of the smaller twin being <10th, 10th to 50th, or >50th percentile. RESULTS: Among the 10,683 pairs of twins who were studied, the respective proportions of the three groups were 62.4%, 32.9%, and 4.7%. The neonatal mortality rate was significantly higher among pairs in which the smaller twin weighed <10th birth weight percentile (29. vs 11.1 and 11 per 1000; odds ratio, 2.7; 95% CI, 1.3, 5.7). This difference results from the higher mortality rates among the smaller but not among the larger twins. CONCLUSION: Severely discordant twin pairs in whom the smaller twin is also small for gestational age are at an increased risk of neonatal death. Identification of this group is an imperative step in the management of birth weight discordance in twin gestations. PMID- 14749656 TI - Obstetric patient satisfaction: asking patients what they like. AB - OBJECTIVE: This study was undertaken to determine pertinent attributes of women's hospital experience related to the delivery of their children and to use open ended responses from women to develop a taxonomy for classifying patient satisfaction in obstetrics. STUDY DESIGN: By using clinimetric methods, we interviewed 67 obstetric patients during their postpartum hospital stays, asking open-ended questions about their satisfaction with care. Responses were transcribed, arranged into distinct groups, and organized as a taxonomy of patient satisfaction. RESULTS: The final taxonomy derived from patient responses was divided into six main axes related to physicians, nurses, other staff, special services, hospital attributes, and personal focus; a total of 51 individual items were identified related to patient satisfaction. These items have face validity, and many are not routinely included in assessments of patient satisfaction. CONCLUSION: A simple strategy of using open-ended questions leads to a clinically relevant and easily understood classification scheme for patient satisfaction with in-hospital obstetric services. PMID- 14749657 TI - Outcomes after expectant management of extremely preterm premature rupture of the membranes. AB - OBJECTIVE: This study was undertaken to assess contemporary outcomes in pregnancies managed expectantly after extremely preterm premature (< or =24 weeks) premature rupture of the membranes (EPPROM). STUDY DESIGN: We queried antepartum and ultrasound databases for patients with EPPROM. Data on pregnancy outcome and short-term neonatal outcomes were collected. RESULTS: Forty-six patients with EPPROM were studied. Patients were hospitalized at 24 weeks' gestation and given antibiotics and antenatal steroids. Median gestational age at PPROM was 22.0 weeks (range 16.9-24 weeks); 43 (93%) elected expectant management, 2 of whom later had an intrauterine fetal death. Median latency period to delivery was 13 days (range 0-96 days), with mean gestational age at delivery of 25.8+/-3.4 weeks. Overall survival was 47% (27 of 57 infants), after a median hospital stay of 71 days (range 17-209 days). Ten (37%) of the survivors have serious sequelae. CONCLUSION: Although significant pregnancy prolongation after previable PPROM occurs in many cases, neonatal outcomes remain poor. PMID- 14749658 TI - Resistance exercise decreases the need for insulin in overweight women with gestational diabetes mellitus. AB - OBJECTIVE: This study examines the effects of circuit-type resistance training on the need for insulin in women with gestational diabetes mellitus. STUDY DESIGN: Thirty-two patients with gestational diabetes mellitus were randomly assigned either to a group that was treated with diet alone or to a group that was treated with diet plus resistance exercise. RESULTS: The number of women whose condition required insulin therapy was the same, regardless of treatment. However, a subgroup analysis that examined only overweight women (prepregnant body mass index, >25 kg/m(2)) showed a lower incidence of insulin use in the diet-plus exercise group (P<.05). Women in the diet-plus-exercise group were prescribed less insulin (P<.05) and showed a longer delay from diagnosis to the initiation of insulin therapy (P<.05), compared with the diet-alone group. CONCLUSION: Resistance exercise training may help to avoid insulin therapy for overweight women with gestational diabetes mellitus. PMID- 14749659 TI - Side-to-side differences in transcranial Doppler parameters in normotensive and preeclamptic pregnant women. AB - OBJECTIVE: This study was undertaken to investigate whether women with preeclampsia demonstrate larger side-to-side velocity differences in the middle cerebral artery (MCA) compared with normotensive pregnant women. STUDY DESIGN: Forty-one preeclamptic women and 50 normotensive pregnant women were studied during the third trimester. Transcranial Doppler ultrasound was used to measure peak, end-diastolic, and mean velocities in both MCAs. An asymmetry index was calculated as 100 x Rt-Lt//(Rt+Lt)/2, for each of the following parameters: mean velocity (Vm), pulsatility index (PI), and cerebral perfusion pressure (CPP). Student t test, Pearson correlation, and regression analysis were used. Significance was taken as P<.05. RESULTS: Both normotensive and preeclamptic pregnant women showed good correlation between Rt and Lt MCA Vm (R>0.8, P<.0001), PI (R>0.6, P.0001), and CPP (R>0.8, P<.0001). There were no differences in the asymmetry index for Vm, PI, or CPP between the two groups. CONCLUSION: Preeclampsia does not appear to induce greater side-to-side velocity differences in the MCA distribution. PMID- 14749660 TI - Cardiac autonomic modulation in normal, high-risk, and in vitro fertilization pregnancies during the first trimester. AB - OBJECTIVES: The purpose of this study was to test the adaptation of autonomic modulation of heart rate in high-risk or in vitro fertilization (IVF) pregnancies during the first trimester. STUDY DESIGN: Thirty-three pregnant women were studied between 6.0 and 12.5 weeks of gestation and were divided into three groups: normal (n=17), high-risk (n=7), and IVF pregnancies (n=9), together with 9 nonpregnant women of comparable age. All subjects underwent a short-term continuous electrocardiographic recording to measure short-term heart rate variability (HRV). RESULTS: Average values of mean R-R interval, total power, and low-frequency (LF) component were similar in nonpregnant and normally pregnant women. The high-frequency (HF) component was only slightly increased in normal pregnant women but no difference was observed in LF/HF ratio. High-risk and IVF pregnancies were characterized by a significant increase in LF component in comparison to normal pregnancies. In pregnancies that had obstetric complications, signs of abnormal autonomic modulation of the sinus node were particularly evident. CONCLUSION: Short-term analysis of HRV in high-risk and IVF pregnancies was significantly different from that observed in normal pregnancies. These differences were more significant in those pregnancies later complicated by an adverse outcome, suggesting an early origin of these pathologic conditions. PMID- 14749661 TI - Eating disorder that was diagnosed before pregnancy and pregnancy outcome. AB - OBJECTIVE: The purpose of this study was to determine the association of an eating disorder that was diagnosed before pregnancy and a preterm delivery and/or the delivery of a low-birth-weight or small-for-gestational-age infant. STUDY DESIGN: This was a register-based follow-up study. We included 302 women who were hospitalized with an eating disorder before pregnancy who were delivered of 504 children and 900 control subjects who were delivered of 1552 children. The association of eating disorders, birth weight, and gestational age was assessed by bivariate and multivariate analyses. RESULTS: The risk of a low-birth-weight infant was twice as high in women with a previous eating disorder compared with women with no such disorder (odds ratio, 2.2; 95% CI, 1.4-3.2). The risk of preterm delivery and a small-for-gestational-age infant was increased to 70% and 80% (odds ratio, 1.7 [95% CI, 1.1-2.6]; odds ratio, 1.8 [95% CI, 1.3-2.4]), respectively. CONCLUSION: Women who were hospitalized for an eating disorder that was diagnosed before pregnancy were at increased risk of impaired pregnancy outcome. PMID- 14749662 TI - Thyrotoxicosis and heart failure that complicate pregnancy. AB - OBJECTIVE: When untreated, Graves' thyrotoxicosis has profound cardiovascular effects, although it rarely causes heart failure in otherwise healthy patients. Preliminary observations suggest that pregnant women are the exception. To further elucidate this association, we studied both immediate and long-term outcomes in women who had thyrotoxicosis and heart failure during pregnancy. STUDY DESIGN: We reviewed clinical outcomes of pregnant women with Graves' disease and heart failure at our institution from 1974 through 2001. Women with other underlying heart disease were excluded. A standardized antithyroid regimen and serial echocardiography and/or chest radiography were performed. RESULTS: The 13 women with thyrotoxicosis and heart failure were either noncompliant with antithyroid therapy or had no medical care during pregnancy. Six women had heart failure before fetal viability; decompensation was precipitated by hemorrhage, sepsis, or both. The other 7 women were in the last trimester when heart failure developed; in 4 women, the heart failure was precipitated by severe preeclampsia eclampsia and in 2 women was precipitated by sepsis. Overall, 11 of 13 women had an underlying obstetric event. In follow-up of 11 women from 2 to 25 years, resolution of cardiomyopathy was confirmed after successful treatment of thyrotoxicosis. CONCLUSION: Normal pregnancy mimics and amplifies some of the hyperdynamic cardiovascular changes that are caused by thyrotoxicosis. When they occur simultaneously, there is usually a compensated high-output state. In some women, however, common pregnancy complications that include hemorrhage with associated anemia, sepsis, and severe preeclampsia-eclampsia will precipitate heart failure. The immediate treatment of heart failure and the correction of precipitating pregnancy factors usually results in good outcome. Long-term follow up confirmed that thyrotoxic cardiac dysfunction is reversible with successful antithyroid therapy. PMID- 14749663 TI - Frequency of infant adverse events that are associated with citalopram use during breast-feeding. AB - OBJECTIVE: The purpose of this study was to determine the frequency of infantile adverse events from exposure through breast-feeding to maternal citalopram therapy. STUDY DESIGN: This was a prospective, observational cohort study. Women who were breast-feeding were placed in three groups on the basis of citalopram use: group 1 consisted of 31 women who were depressed and were undergoing citalopram therapy, group 2 consisted of 12 women who were depressed but were not undergoing citalopram therapy, and group 3 consisted of 31 healthy women who were matched to group 1 by maternal age and parity. Data collection included infant feeding method, medication use, and adverse events. RESULTS: There was no statistically significant difference in the rate of adverse events in the three groups (3/31 events, 0/12 events, and 1/31 events in groups 1, 2, and 3, respectively). The average dose of citalopram that was used in group 1 was 25.3+/ 11.4 mg per day (range, 10-60 mg/d). CONCLUSION: To our knowledge, this is the first prospective, controlled study to examine the safety of citalopram during breast-feeding, which should be continued during maternal citalopram therapy. PMID- 14749664 TI - Effect of an oxytocin receptor antagonist and rho kinase inhibitor on the [Ca++]i sensitivity of human myometrium. AB - OBJECTIVE: The purpose of this study was to assess the possible mechanism and effect of oxytocin receptor on the sensitization of the contractile proteins to [Ca(++)](i). STUDY DESIGN: Myometrial strips were prepared from biopsy specimens that had been taken at term (37-40 weeks of gestation) cesarean delivery from 22 women, before the onset of labor. Simultaneous measurements of [Ca(++)](i) and tension were performed on spontaneously contracting strips. The effects of a specific oxytocin receptor antagonist, L371,257, and a rho kinase inhibitor, Y 27632, on the [Ca(++)](i) and tension transients were determined. RESULTS: Application of L371,257 (90 nmol/L) to spontaneously contracting muscle strips reduced peak tension to 43%+/-12% of its original value without affecting peak [Ca(++)](i) (105%+/-15%). More tension is developed at each [Ca(++)](i) during the falling phase, relative to the rising phase of each spontaneous contraction; this asymmetric [Ca(++)](i)-tension relationship was abolished by L371,257. The subsequent application of oxytocin (3 micromol/L) reversed the effects of the antagonist on tension. An inhibitor of rho kinase (Y-27632) caused a concentration-dependent inhibition of spontaneous contractions without affecting the underlying Ca(++) signals. The addition of oxytocin (1 nmol/L) to spontaneously active myometrium increased both the duration of the Ca(++) signal and the sensitivity of the contractile machinery to [Ca(++)](i); only the latter effect was prevented by Y-27632. CONCLUSION: Oxytocin independently regulates the duration of the periodic Ca(++) signals and the sensitivity of the contractile machinery to Ca(++). The latter is likely to be mediated by rho kinase, which is essential for the effective coupling of increases in [Ca(++)](i) to tension. PMID- 14749665 TI - Induction of labor with RU 486 (mifepristone) in relaxin-deficient rats: antepartum administration of relaxin facilitates delivery and increases pup survival. AB - OBJECTIVE: This study was conducted to determine whether antepartum administration of relaxin improves RU 486-induced delivery at term in rats that lack circulating endogenous relaxin. STUDY DESIGN: Pregnant rats were modified two ways to obtain circulating levels of relaxin and progesterone that resemble those of pregnant humans: relaxin was immunoneutralized throughout the second half of the 23-day pregnancy and high progesterone levels were sustained until term by inserting progesterone implants on day 20. Porcine relaxin was administered subcutaneously from 8 AM on day 20 until delivery. Labor was induced by administering RU 486 subcutaneously at 4 AM on day 22. RESULTS: After induction of labor with RU 486, labor and delivery were faster, and the incidence of live births was higher when rats were also administered relaxin during the antepartum period. CONCLUSION: Antepartum administration of relaxin in combination with RU 486 has beneficial effects on delivery in relaxin-deficient rats. PMID- 14749666 TI - Immunologic properties of human fetal mesenchymal stem cells. AB - OBJECTIVE: Mesenchymal stem cells (MSCs) can be isolated from adult bone marrow and fetal liver. We investigated the immunologic properties of undifferentiated and differentiated human fetal MSCs. STUDY DESIGN: Expression of HLA class I and II was investigated by flow cytometry and Western blot on undifferentiated fetal MSC and after in vitro differentiation to adipocytes and osteocytes. Alloreactivity was studied after adding fetal MSCs to allogeneic lymphocytes in mixed lymphocyte cultures. RESULTS: Fetal MSCs expressed HLA class I but not HLA class II. The presence of interferon gamma (IFN-gamma) in the growth medium for 2 days initiated the intracellular synthesis of HLA class II, but 7 days of exposure was required for cell surface expression. Neither undifferentiated nor differentiated fetal MSCs induced proliferation of allogenic lymphocytes. Fetal MSCs treated with IFN-gamma suppressed alloreactive lymphocytes. CONCLUSION: Undifferentiated and differentiated fetal MSCs do not elicit alloreactive lymphocyte proliferation. The results suggest that fetal MSCs have potentials for allogenic transplantation. PMID- 14749667 TI - Effects of thyrotropin-releasing hormone on human myometrium and umbilical vasculature in vitro. AB - OBJECTIVE: The purpose of this study was to investigate the direct effects of thyrotropin-releasing hormone on isolated human myometrium that was obtained during pregnancy and on human umbilical vasculature in vitro. STUDY DESIGN: Isolated human myometrial strips were dissected from biopsy specimens that were obtained at elective cesarean delivery and suspended for isometric recording under physiologic conditions. The effects of cumulative additions of thyrotropin releasing hormone (10(-9)-10(-4) mol/L) on oxytocin-induced myometrial contractility were evaluated. The effects of thyrotropin-releasing hormone (10( 9)-10(-4) mol/L) on umbilical vessel (artery and vein) resistance in vitro were investigated with the use of isolated ring preparations. RESULTS: Thyrotropin releasing hormone exerted a significant concentration-dependent relaxant effect on pregnant human myometrial tissue, which ranged from 3.54% (10(-9) mol/L, P=.935) to a net cumulative total of 21.06% (10(-4) mol/L, P<.001). Thyrotropin releasing hormone also exerted a concentration-dependent relaxant effect on human umbilical vasculature that ranged from 12.51% (10(-9) mol/L, P=.994) to a net cumulative total of 23.27%+/-4.87% (SEM, 10(-4) mol/L, P<.01) in umbilical artery. For umbilical vein, the relaxant effect ranged from 1.80% (10(-9) mol/L, P=.998) to a net cumulative total of 14.64% (10(-4) mol/L, P<.009). CONCLUSION: Thyrotropin-releasing hormone exerts a significant relaxant effect in human myometrium and in human umbilical vasculature and highlights a potential physiologic role for this neuropeptide in these tissues. These findings have clinical implications for the therapeutic use of thyrotropin-releasing hormone antenatally. PMID- 14749668 TI - Neutrophils are stimulated by syncytiotrophoblast microvillous membranes to generate superoxide radicals in women with preeclampsia. AB - OBJECTIVE: This study was undertaken to determine whether syncytiotrophoblast microvillous membranes (STBMs) stimulate maternal neutrophils to produce superoxide radicals in women with preeclampsia. STUDY DESIGN: Serum levels of tissue polypeptide antigen (TPA), which is a marker for STBM, were measured in 25 nulliparous women (10 with mild preeclampsia, 6 with severe preeclampsia, and 9 controls). Superoxide production by maternal neutrophils from cases and controls and by donor neutrophils cocultured with the STBMs from cases and controls was measured spectrophotometrically by reduction of ferricytochrome C. RESULTS: Maternal TPA levels were significantly greater among cases than controls (P=.005). Superoxide production by maternal neutrophils and donor neutrophils cultured with STBM from cases of preeclampsia was greater than controls (P values.006 and.019, respectively), and dose-response relationships were observed. Superoxide production by maternal leukocytes was correlated with superoxide induction by STBMs in culture (P=.007). CONCLUSION: STBMs in maternal blood induce neutrophils to generate superoxide radicals that may cause endothelial dysfunction in women with preeclampsia. PMID- 14749669 TI - Chronic prenatal exposure to carbamazepine and perinatal outcomes of C3H/He mice. AB - OBJECTIVES: This study was undertaken to determine a daily therapeutic dose of carbamazepine and to measure its effect on reproductive performance and perinatal outcomes of mice. STUDY DESIGN: Adult C3H/He mice were given carbamazepine in rodent chow in either a 0.25% or a 1.0% mixture. Comparisons between doses included nongravid weight change, plasma drug steady-state concentrations, and response to a maximal electroshock seizure test. The strain was then fed either the preferred dose of carbamazepine or a placebo 1 week before starting to mate and throughout gestation to compare reproductive performance and offspring early development. RESULTS: Mice who ate the 0.25% carbamazepine mixture displayed no evoked seizure activity and, in contrast to the 1.0% mixture, did not lose weight. This daily dose of 542+/-35 mg/kg produced a trough steady-state plasma concentration that was consistent with a protective threshold in humans. Differences from placebo controls were not statistically significant for the number of cycles necessary to conceive or for the duration of gestation. The litter size, survival rates, birth weights, weight gain, and onset of eye openings and teeth eruptions of the pups were not statistically significant between the two groups. CONCLUSION: Long-term prenatal exposure to a subtoxic yet therapeutic dose of carbamazepine did not impair reproductive performance or early growth and development of exposed mice offspring. PMID- 14749670 TI - Fetal membranes act as a barrier for adenoviruses: gene transfer into exocoelomic cavity of rat fetuses does not affect cells in the fetus. AB - OBJECTIVES: In utero gene therapy has a potential to correct genetic disorders before the first clinical symptoms appear. Our aim was to examine whether the exocoelomic cavity between amniotic and chorionic membranes offers a minimally invasive route for gene transfer to the fetus during early pregnancy. STUDY DESIGN: We injected lacZ-adenovirus (4 x 10(9) pfu) during open surgery into the exocoelomic cavity of rat fetuses (n=50) and analyzed the fetuses and rat dams for transgene expression with X-gal staining and polymerase chain reaction. RESULTS: Giant cells around Reichert's membrane, the outermost extraembryonic membrane in rodents, were transduced; but no transduction was observed in the cells of the fetuses or rat dams. CONCLUSION: In rodents, the exocoelomic cavity does not offer a route for gene transfer into the fetus. It was concluded that fetal membranes act as a barrier that prevents adenoviral particles from passing between embryonic cavities. PMID- 14749671 TI - The presence and function of phosphodiesterase type 5 in the rat myometrium. AB - OBJECTIVE: Cyclic nucleotide phosphodiesterases (PDEs) are a diverse enzyme group with multiple regulatory properties and wide tissue distribution. Such activity includes cyclic adenosine (cAMP) and guanosine monophosphate (cGMP) breakdown. The type 5 isoform (PDE-5, cGMP specific) is the target of specific antagonists (ie, sildenafil, Viagra). We tested the hypothesis that PDE-5 is present in rat myometrium and modulates myometrial activity. STUDY DESIGN: Full-thickness uterine wall was collected from nonpregnant (n=3) and pregnant Sprague-Dawley rats on days 10 (n=4), 17 (n=6), 22 nonlabor (n=5), and 22 during term labor (TL, n=4). Preterm labor (PTL, n=3) was induced in some animals on day 16 with 15 mg/kg mifepristone (RU 486). Tissue samples were prepared for Western blotting using a monoclonal antibody against rodent PDE-5. In a second series, cumulative doses of sildenafil (0.005, 0.05, 0.5, 5 mg/kg, intraperitoneal) were administered and the effect on uterine contractility recorded in vivo during term (TL, n=7) and preterm labor (PTL, n=6). Saline solution-injected rats provided temporal control. Uterine contractility was estimated from intrauterine pressure (IP) measured electronically with a sensor tip pressure catheter. Heart rate was recorded simultaneously using electrodes attached to the chest and connected to the same data acquisition system. RESULTS: PDE-5 immunoreactivity was present in the nonpregnant rat uterus and at all gestational times studied, although the expression was unaffected by either pregnancy or the state of labor (preterm or term). A dominant antibody-specific band was identified at 86 kd in the uterine samples, contrasting with lung where the 100-kd PDE-5 isoform was most abundant. Two additional lower molecular weight (55 and 32 kd) bands were also identified as antibody specific. Despite the lack of change in PDE-5 during pregnancy, sildenafil reduced IP during TL and PTL beginning at 0.5 mg/kg. The highest dose of sildenafil reduced IP during both TL and PTL by 45% and 59% of baseline, respectively (two-way analysis of variance, P<.01). This effect was not accompanied by changes in heart rate. CONCLUSION: PDE-5 is constitutively present in the rat uterine wall. There was no observed change in the PDE-5 protein expression throughout pregnancy. In contrast to the lung, the uterus expresses an 80-kd PDE-5 isoform. Sildenafil in pharmacologic doses inhibits mechanical uterine activity and might be of benefit if selectively used for treatment of preterm labor. PMID- 14749673 TI - Three-dimensional sonographic measurement of normal fetal brain volume during the second half of pregnancy. AB - OBJECTIVES: This study was undertaken to develop a three-dimensional (3D) ultrasound method of measuring fetal brain volume. STUDY DESIGN: Serial 3D sonographic measurements of fetal brain volume were made in 68 normal singleton pregnancies at 18 to 34 weeks of gestation. A comparison was made with fetal brain volume estimates from two-dimensional (2D) sonographic measurement of head circumference and published postmortem fetal brain weights. RESULTS: Coefficient of variation for fetal brain volume (3D) caused by differences between repeated tests was 10.2% and between analyses of the same recorded volume 2.2%. Median brain volume increases from 34 mL at 18 weeks to 316 mL at 34 weeks. Median brain weight represented approximately 15% of total fetal weight. The 3D ultrasound derived brain weight is larger than postmortem brain weight. However, this is not so for brain weight derived from total fetal weight at autopsy. A good agreement between 3D and 2D brain volume was found. CONCLUSION: Sonographic measurement of fetal brain volume demonstrated an acceptable intraobserver variability and a nearly 10-fold increase during the second half of gestation. PMID- 14749674 TI - Trichomoniasis: under control or undercontrolled? AB - Aside from human papillomavirus, trichomoniasis is the most common sexually transmitted infection in the United States today, yet it has long been regarded as a sexually transmitted infection of minor importance. Medical opinion has traditionally held that it plays little role in health complications in women, and it is rarely seen in men. However, evidence has recently accumulated implicating Trichomonas vaginalis as a contributor to a variety of adverse outcomes among both sexes. Among both women and men, the association of T vaginalis with human immunodeficiency acquisition and transmission has been shown in multiple studies. Among women, trichomoniasis may play a role in development of cervical neoplasia, postoperative infections, and adverse pregnancy outcomes and as a factor in atypical pelvic inflammatory disease and infertility. Among men, trichomoniasis has emerged as a cause of nongonoccocal urethritis and as contributing to male factor infertility. As evidence continues to accumulate, the time has come to increase diagnostic efforts beyond traditional sexually transmitted disease clinic settings. PMID- 14749675 TI - Endometriosis--the consequences of neurologic dysfunction. PMID- 14749677 TI - External validation of a new logistic model in the diagnosis of ovarian cancer. PMID- 14749679 TI - Impact of obesity on spontaneous abortion. PMID- 14749680 TI - Best practices for insertion of central venous catheters in intensive-care units to prevent catheter-related bloodstream infections. PMID- 14749681 TI - 1-alpha,25-Dihydroxyvitamin D3 regulates inducible nitric oxide synthase messenger RNA expression and nitric oxide release in macrophage-like RAW 264.7 cells. AB - The expression of inducible nitric oxide synthase (iNOS) expression and release of nitric oxide (NO) from macrophages are markedly increased in granulomatous infections. Activation of macrophages 1alpha-hydroxylase results in an increase of 1alpha,25-dihydroxyvitamin D(3) [1,25(OH)(2)D(3)]. However, the significance of this increased production is not completely understood. In this study, we analyzed 1,25(OH)(2)D(3) and NO production in patients with tuberculosis infection and hypercalcemia and used lipopolysaccharide (LPS) to stimulate RAW 264.7 cells in an attempt to assess iNOS expression and gaseous NO production regulated by 1,25(OH)(2)D(3). Peroxynitrite (OONO(-)) production and lactate dehydrogenase activity were also examined. Without additional stimulation, peripheral-blood mononuclear cells (PBMCs) from patients with tuberculosis converted more 25-hydroxyvitamin D(3) to 1,25(OH)(2)D(3) than did those from normal controls. These PBMCs released less NO than did those from control subjects, at baseline and in the stimulated state. We found that 1,25(OH)(2)D(3) dose-dependently inhibited iNOS messenger RNA expression of the LPS-stimulated RAW 264.7 cells and also significantly reduced the gaseous NO release and OONO(-) production. Paralleling the 1,25(OH)(2)D(3)-induced inhibition of NO release were reductions in OONO(-) and LDH production. In conclusion, 1,25(OH)(2)D(3) inhibited iNOS expression and reduced NO production by LPS-stimulated macrophages in the range of physiological doses. Inhibition of the NO surge was coupled with a reduction in OONO(-) and LDH production. Increased 1,25(OH)(2)D(3) production and decreased release of NO from the PBMCs of patients with tuberculosis and hypercalcemia were also noted. We propose that 1,25(OH)(2)D(3) production by macrophages may protect themselves against oxidative injuries caused by the NO burst. In the case of tuberculosis infection, increased 1,25(OH)(2)D(3) synthesis may further contribute to the development of an unwanted phenomenon hypercalcemia. PMID- 14749682 TI - Urinary fatty acid-binding protein as a new clinical marker of the progression of chronic renal disease. AB - Previous studies have indicated that in massive proteinuria, free fatty acids (FFAs) bound to albumin were overloaded in the proximal tubule and exacerbated tubulointerstitial damage. Liver-type fatty acid-binding protein (L-FABP) is an intracellular carrier protein of FFAs that is expressed in the proximal tubule of human kidney. We sought to evaluate urinary L-FABP as a clinical marker in chronic renal disease. Urinary L-FABP was measured in patients with nondiabetic chronic renal disease (n = 120) with the use of a newly established ELISA method. We then monitored these patients for 15 to 51 months. Clinical data were analyzed with multivariate analysis. Urinary L-FABP was correlated with urinary protein, urinary alpha(1)-microglobulin, and serum creatinine concentrations. Urinary L FABP at the start of follow-up (F = 17.1, r =.36, P <.0001) was selected as a significant clinical factor correlated with the progression rate, defined as a slope of a reciprocal of serum creatinine over time. We next selected the patients with mild renal dysfunction (n = 35) from all 120 patients and divided them into 2 groups according to progression rate: the progression group (n = 22) and the nonprogression group (n = 13). Serum creatinine and urinary protein concentrations and blood pressure at the start of follow-up were higher in the progression group than in the nonprogression group, although we detected no significant difference between the 2 groups. Urinary L-FABP was significantly higher in the former group than in the latter (P <.05). The results showed that urinary L-FABP reflected the clinical prognosis of chronic renal disease. Urinary L-FABP may be a clinical marker that can help predict the progression of chronic glomerular disease. PMID- 14749683 TI - Acute oxidative stress induces peritoneal hyperpermeability, mesothelial loss, and fibrosis. AB - We explored the acute and long-term effects of short-lived, intense oxidative stress on peritoneal permeability and structure, induced with intraperitoneal injection of the oxidant agent deoxycholate, in rats. Ten minutes after the experimental intervention, peritoneal dialysis, performed over an exposure time of 60 minutes, revealed an increased urea dialysate/plasma ratio, greater glucose absorption, increased albumin losses in the effluent dialysate, and a reduced ultrafiltration rate. Mesothelial-cell imprints taken from the anterior liver surface indicated a substantially decreased density in the cell population. After the recovery period of 30 days, all alterations were still evident. Additionally, macroscopic and histologic observations made at this time interval detected peritoneal fibrosis and sclerosis, characterized by peritoneal adhesions, wrapping of intestinal loops, and the presence of a layer of fibrous tissue dressing the cavitary aspect of the liver peritoneal envelope. This report describes a reproducible experimental model of peritoneal fibrosis induced by acute oxidative injury. On the basis of these findings, it may be speculated that functional and structural alterations observed in patients are related to long term continuous exposure of the monolayer to oxidative injury resulting from the high concentrations of d-glucose present in peritoneal dialysis solutions. PMID- 14749684 TI - Tissue angiotensin II in the regulation of inflammatory and fibrogenic components of repair in the rat heart. AB - Quantitative in vitro autoradiography has identified high density ACE and AT(1) receptor binding at sites of cardiac injury in the adult rat, implicating Ang II, generated de novo at these sites (tissue Ang II) in contributing to repair. This hypothesis remains to be tested. In the study reported here we used a time dependent rat model of cardiac injury wherein plasma levels of renin and Ang II are chronically suppressed by means of continuous treatment with aldosterone (0.75 microg/h) and 1% dietary NaCl. To further address a role for tissue Ang II in tissue repair, we administered oral valsartan (10 mg/kg/day) in combination with aldosterone/NaCl. On days 20 and 30 of each regimen, hearts were examined. In coronal sections, we assessed transcription factor NFkappaB activation (RelA subunit), inflammatory-cell infiltration and appearance of myofibroblasts by immunohistochemistry; mRNA expression of several inflammatory (NFkappaB-related) and fibrogenic (type I collagen) mediators of repair, using quantitative in situ hybridization; and ACE binding density, detected with quantitative in vitro autoradiography. Blood pressure was measured with a tail cuff. Untreated age- and sex-matched rats served as controls. On day 20, we found no evidence of cardiac injury, inflammation, or repair with aldosterone/NaCl treatment, with or without valsartan. In contrast, on day 30 of aldosterone/NaCl treatment, inflammatory cells and alpha-SMA-positive myofibroblasts colocalized with high-density ACE binding and histochemical evidence of fibrillar collagen accumulation at sites of microscopic scarring and perivascular fibrosis of intramyocardial coronary arteries that appeared in both right and left ventricles. The activation of NFkappaB and the increased mRNA expression of ICAM-1, MCP-1, TNF-alpha, TGF beta(1), PAI-1, and type I collagen were also observed at these sites. Expression of vascular cell adhesion molecule-1 was unchanged. Valsartan significantly reduced (P <.01) the expression of these mediators and attenuated the expression of MCP-1. It reduced microscopic evidence of tissue damage and the extent of fibrosis. Blood pressure was increased in aldosterone-treated rats on days 20 and 30; this increase was suppressed by valsartan. We thus show that in this rat model of long-term aldosterone/NaCl administration, in which circulating Ang II is suppressed, AT(1) receptor-mediated actions of tissue Ang II are involved in regulating the expression of mediators of repair at vascular and nonvascular sites of cardiac injury, thereby implicating autocrine/paracrine properties of tissue Ang II in inflammatory and healing responses. PMID- 14749685 TI - Role of prostaglandins in maintaining gastric mucus-cell permeability against acid exposure. AB - Regulation of gastric epithelial permeability is important in the protection of the gastric mucosa from secreted acid. However, the mechanism(s) for this regulation in gastric mucus cells remains unknown. In this study, we evaluated gastric epithelial-cell permeability in response to acid exposure by monitoring trans-epithelial electrical resistance (TEER) and paracellular permeability with carbon 14-labeled mannitol. We also examined the role of prostaglandins on gastric epithelial permeability. Rat gastric epithelial cells (RGM-1) were plated on 8-microm-pore tissue-culture inserts. Cells were exposed to solutions of differing pH (3-7.4), with and without the nonsteroidal antiinflammatory drug (NSAID) indomethacin (10(-7) mol/L), for 60 to 120 minutes. Transepithelial permeability was measured on the basis of TEER and the diffusion rate of [(14)C]mannitol. Prostaglandin E(2) (PGE(2)) was administered in some experiments with NSAIDs. After acid exposure (pH 3.0-5.0), TEER rapidly and significantly increased, peaking in 5 minutes. Diffusion of [(14)C]mannitol was blocked during the period when TEER increased. Pretreatment with the cyclooxygenase (COX) inhibitor indomethacin blocked the rapid acid-induced increase in TEER. A specific COX-2 inhibitor had no effect on this rapid increase in TEER. The blockade by indomethacin was eliminated by the addition of PGE(2). These findings suggest that when gastric-surface mucus cells are exposed to acid, gastric epithelial permeability decreases rapidly to inhibit acid back-diffusion. Prostaglandins play an important role in this protective response to acid exposure. COX inhibitors such as indomethacin may inhibit the regulation of epithelial permeability by reducing the concentration of PGE(2). PMID- 14749686 TI - Immunoassay for wild-type protein in lymphocytes predicts germline mutations in patients at risk for hereditary colorectal cancer. AB - Using colorectal cancer (CRC) as an example, we present the hypothesis that quantitative immunoassays for wild-type (full-length) proteins can be used to identify carriers of traits for hereditary diseases. In the case of hereditary CRC, this involves identifying individuals with germline mutations in a mismatch repair (MMR) gene (mainly hMSH2 or hMLH1) or in the adenomatous polyposis coli (APC) gene. Because expression of wild-type protein should reflect wild-type gene dosage, we predicted that individuals harboring a germline mutation will have a reduction of approximately 50% in expression in lymphocytes of the corresponding full-length protein. In this pilot study, we tested lymphoblastoid cell lines that had been established from controls and individuals with, or at high risk for, hereditary CRC: 9 lines from healthy, unaffected individuals; 4 from affected members in familial adenomatous polyposis families (with known germ-line APC mutation); 42 from CRC patients in our Familial CRC Registry (increased risk of hereditary nonpolyposis colon cancer as assessed by family history, age at adenoma or carcinoma diagnosis, and other clinical criteria). For MSH2 and MLH1 we used western blots; for APC we used immunoprecipitation. All familial adenomatous polyposis lines had about 50% less immunoprecipitable full-length APC protein. Some cell lines (7 of 42) from Familial CRC Registry patients showed on western blots a reduction (mean 46%) in either MSH2 or MLH1 (relative to the other protein). All 7 subsequently were proved to contain a germline MMR mutation. We conclude that (1) because most of the expected CRC-causing germ line mutations are truncation-causing, immunoassays for wild-type protein should be able to identify most individuals with hereditary CRC-causing traits; (2) these assays, which are more practical and inexpensive than current mutation-detecting tests for hereditary CRC traits, have the potential for commercial development into broad-based population screens of high-risk patients and their families and the potential to save both lives and health-care dollars; (3) this strategy may be useful for other hereditary cancers and even other hereditary diseases; (4) our approach has the potential to greatly benefit public-health programs for cancer control. PMID- 14749687 TI - 250 years of controlled clinical trials: where it all began. PMID- 14749688 TI - Drug interactions with herbal products and grapefruit juice: a conference report. PMID- 14749689 TI - Polymorphisms in human MDR1 (P-glycoprotein): recent advances and clinical relevance. AB - Drug transporters are increasingly recognized to be important to drug disposition and response. P-glycoprotein, the encoded product of the human MDR1 (ABCB1) gene, is of particular clinical relevance in that this transporter has broad substrate specificity, including a variety of structurally divergent drugs in clinical use today. Moreover, expression of this efflux transporter in certain tissue compartments such as the gastrointestinal tract and brain capillary endothelial cells limits oral absorption and central nervous system entry of many drugs. Recently, a number of single-nucleotide polymorphisms (SNPs) in MDR1 have been identified. An increasing number of studies have also implicated certain commonly occurring SNPs in MDR1 in problems including altered drug levels and host susceptibility to diseases such as Parkinson's disease, inflammatory bowel disease, refractory seizures, and CD4 cell recovery during human immunodeficiency virus therapy. However, in many such cases, the reported effects of MDR1 SNPs have been inconsistent and, in some cases, conflicting. In this review SNPs in MDR1 in relation to population frequencies, drug levels, and phenotypes are outlined. In addition, issues relating to MDR1 haplotypes, environmental factors, and study design, as potential confounding factors of the observed MDR1 polymorphism effect in vivo, are also discussed. PMID- 14749691 TI - Mechanisms of beta-adrenergic receptor-mediated venodilation in humans. AB - OBJECTIVES: Recent studies suggest that stimulation of beta-adrenergic receptors results in both endothelium-dependent and endothelium-independent venodilation, but results of former studies are inconsistent. This study was designed to elucidate the underlying mechanisms of isoproterenol (INN, isoprenaline)-induced venodilation by investigation of dorsal hand vein responses. METHODS: In phenylephrine-constricted veins, isoproterenol (2-514 ng/min) was infused with and without oral pretreatment with 1 g acetylsalicylic acid (n = 7) or 5 mg of the selective beta(1)-adrenergic receptor antagonist bisoprolol (n = 7). In addition, isoproterenol was coinfused with the nitric oxide inhibitor N(G) monomethyl-l-arginine (l-NMMA) (6.3 micromol/min [n = 6]), with selective blockers of calcium (Ca(++))-dependent potassium (K(+)) channels (tetraethylammonium, 300 microg/min [n = 6]) and adenosine triphosphate (ATP) sensitive K(+) channels (glyburide [INN, glibenclamide], 20 microg/min [n = 6]) or with the cyclic guanosine monophosphate inhibitor methylene blue (13 microg/min [n = 6]). Finally, L-NMMA was coinfused with potassium chloride (20 mmol/L) to inhibit hyperpolarization (n = 6). RESULTS: Isoproterenol induced dose dependent venodilation to 67.4% +/- 6.8%. Oral pretreatment with bisoprolol (P =.340) or acetylsalicylic acid (P =.760) did not affect isoproterenol-induced venodilation. Coinfusion of isoproterenol and L-NMMA relaxed the veins to the same extent as in the presence of isoproterenol alone. Neither inhibition of ATP sensitive K(+) channels (P =.196) nor blockade of Ca(++)-dependent K(+) channels (P =.640) modulated isoproterenol-induced venodilation. In contrast, methylene blue reduced the maximum response to isoproterenol by about one third (68.5% +/- 4.3% versus 41.7% +/- 5.5%, P =.001). Infusion of L-NMMA alone raised vein size to 38.8% +/- 6.5%, yielding an L-NMMA-sensitive increase of 20% (P =.001), which was antagonized by coinfusion of potassium chloride to 17.1% +/- 6.7% (P =.02). CONCLUSIONS: Isoproterenol dilates human hand veins exclusively via beta(2) adrenergic receptors without involvement of endothelium-derived epoprostenol. Although a contribution of endothelium-derived nitric oxide appears unlikely, the venodilating effect of L-NMMA could have obscured the nitric oxide component of isoproterenol. beta(2)-Adrenergic receptor-mediated dilation is mediated in part by cyclic guanosine monophosphate-dependent mechanisms, whereas ATP- and Ca(++) dependent K(+) channels are not involved, excluding a significant contribution of smooth muscle cell hyperpolarization. In addition, high concentrations of the nitric oxide synthase blocker L-NMMA dilate human hand veins via activation of endothelium-derived hyperpolarizing factors. PMID- 14749692 TI - Abciximab pharmacodynamic model with neural networks used to integrate sources of patient variability. AB - OBJECTIVE: Our objective was to develop a computational model for predicting abciximab-induced inhibition of ex vivo platelet aggregation from the administered dose and readily available patient clinical characteristics by use of a neural network approach. METHODS: A back-propagation neural network was designed to establish the relationship between abciximab dosing, patient clinical history, and effect (inhibition of 20 micromol/L adenosine diphosphate-induced ex vivo platelet aggregation). The neural network was trained by use of data from 8 (out of 47) patients undergoing coronary angioplasty and 30 healthy individuals. Final neuron connection weights were used to evaluate significant patient covariates. The final neural network was validated via (1) predicting the effects of the validation database (remaining 39 patients) and (2) predicting the individual patient doses to achieve 20% of baseline platelet aggregation. RESULTS: The trained neural network successfully captured the complex pharmacodynamic profiles of abciximab without specifying a structural model and identified several patient covariates that significantly contribute to establishing the abciximab dose-effect relationship, including stable angina with nitrate treatment, previous myocardial infarction, and smoking. A wide distribution of individual bolus doses of abciximab was predicted, suggesting the potential for dosing individualization while improving the risk of adverse drug events. The mean predicted dose (16.9 mg) was in agreement with the results from a previously published concentration-effect relationship for abciximab (18.9 +/- 2.0 mg). CONCLUSIONS: These findings suggest the usefulness of neural network methods to individualize dosing for drugs with a narrow therapeutic index when real-time measures of drug concentration and effect are unavailable, but future clinical studies are required for prospective validation. PMID- 14749693 TI - Lanreotide effect on splanchnic blood flow in healthy subjects: effect of the rate of infusion. AB - BACKGROUND: Somatostatin is a naturally occurring peptide advocated for the management of hemodynamic complications of chronic liver diseases. The route of administration (bolus application or constant infusion) has been a question of debate. AIM: Our aim was to explore the effects of the somatostatin analog lanreotide, given as a bolus injection or continuous infusion, on food-stimulated hemodynamics in humans. METHODS: Twelve healthy subjects (6 men and 6 women) were studied in a double-blind, double-dummy, randomized, crossover study. After a baseline period of 60 minutes, each subject received either a placebo bolus injection and an intravenous infusion of 100 microg/h lanreotide over a period of 8 hours or a placebo infusion over a period of 8 hours and an 800-microg lanreotide bolus injection. Simultaneously, a liquid test meal (Ensure Plus, 6.3 kJ/mL; Abbott Laboratories, Abbott Park, Ill) was perfused intraduodenally at 3 mL/min over a period of 8 hours. Diastolic blood pressure, heart rate, and superior mesenteric arterial and portal venous volume flows were measured at regular intervals by use of echo-Doppler technology. Plasma lanreotide levels were determined at defined intervals. RESULTS: Lanreotide as a 100-microg/h infusion for 8 hours was bioequivalent with lanreotide as an 800-microg bolus injection (mean area under the plasma concentration-time curve [AUC] extrapolated to infinity [AUC( infinity )], 1844.3 ng.min/L versus 1971.0 ng.min/L; AUC(infinity) ratios, 0.99; confidence interval, 0.95-1.02), and clearance was identical (479.2 mL/min versus 413.4 mL/min, P >.05). As expected, significant differences were observed in maximum plasma concentrations (75.58 ng/mL versus 4.85 ng/mL, P <.001) after infusion and bolus injections, respectively. Lanreotide at 100 microg/h over a period of 8 hours was well tolerated and abolished food-stimulated splanchnic hyperemia in both the superior mesenteric artery and the portal vein (mean AUC above baseline values [AUC(ab)], 37.25 L/min.min and 0.51 L/min.min, respectively). In contrast, the same dose of lanreotide given as a bolus injection only temporarily blunted postprandial hyperemia (mean AUC(ab) for superior mesenteric artery, 251.4 L/min.min, P <.001; mean AUC(ab) for portal vein, 194.95 L/min.min, P <.001), and subjects had significantly more side effects. CONCLUSION: On the basis of tolerability and hemodynamic effects, an intravenous infusion of lanreotide seems superior to a bolus injection of the same dose. PMID- 14749694 TI - Cytochrome P450 1A2 is a major determinant of lidocaine metabolism in vivo: effects of liver function. AB - OBJECTIVES: This study was designed (1) to evaluate the effect of a cytochrome P450 (CYP) 1A2 inhibitor, fluvoxamine, on the pharmacokinetics of intravenous lidocaine and its 2 pharmacologically active metabolites, monoethylglycinexylidide (MEGX) and glycinexylidide (GX), to confirm recent in vitro results indicating that CYP1A2 is the main isoform responsible for lidocaine biotransformation and (2) to assess whether liver function has any influence on the fluvoxamine-lidocaine interaction. METHODS: The study was carried out in 10 healthy volunteers and 20 patients with cirrhosis, 10 with mild (Child grade A) and 10 with severe (Child grade C) liver dysfunction, according to a randomized, double-blind, 2-phase, crossover design. In one phase all participants received placebo for 6 days; in the other phase they received 50 mg fluvoxamine for 2 days and 100 mg fluvoxamine for the next 4 days. On day 6, a 1 mg/kg lidocaine dose was administered intravenously 2 hours after the last dose of fluvoxamine or placebo. Plasma concentrations of lidocaine, MEGX, GX, and fluvoxamine were measured up to 12 hours after lidocaine injection. RESULTS: The effects of fluvoxamine coadministration were dependent on liver function. Lidocaine clearance was decreased on average by 60% (from 12.1 mL/min.kg to 4.85 mL/min.kg, P <.001) in healthy subjects and by 44% (from 9.83 mL/min.kg to 5.06 mL/min.kg, P <.001) in patients with mild liver dysfunction, with proportional increases in terminal half-lives, whereas virtually no effect was produced in patients with severe liver dysfunction (4.21 mL/min.kg versus 3.65 mL/min.kg, P >.05). Analogous effects were observed on MEGX and GX formation kinetics, which were drastically impaired in healthy subjects and patients with mild liver cirrhosis but virtually unaffected in patients with severe cirrhosis. CONCLUSION: CYP1A2 is the enzyme principally responsible for the metabolic disposition of lidocaine in subjects with normal liver function. The extent of fluvoxamine lidocaine interaction decreases as liver function worsens, most likely because of the concomitant decrease in the hepatic level of CYP1A2. These observations indicate that results obtained in healthy subjects cannot be extended a priori to patients with liver dysfunction, but the clinical consequences of inhibition of drug metabolism must also be assessed in such patients. PMID- 14749690 TI - Randomized trial of buprenorphine for treatment of concurrent opiate and cocaine dependence. AB - BACKGROUND: Buprenorphine is a partial mu-opiate agonist and kappa-opiate antagonist with established efficacy in the treatment of opiate dependence. Its efficacy for cocaine dependence is uncertain. This study evaluated buprenorphine for the treatment of concomitant cocaine and opiate dependence. METHODS: Two hundred outpatients currently dependent on both cocaine and opiates were randomly assigned to double-blind groups receiving a sublingual solution of buprenorphine (2, 8, or 16 mg daily, or 16 mg on alternate days, or placebo), plus weekly individual drug abuse counseling, for 13 weeks. The chief outcome measures were urine concentrations of opiate and cocaine metabolites (quantitative) and proportion of urine samples positive for opiates or cocaine (qualitative). Group differences were assessed by use of mixed regression modeling. RESULTS: The target dose of buprenorphine was achieved in 179 subjects. Subjects receiving 8 or 16 mg buprenorphine daily showed statistically significant decreases in urine morphine levels (P =.0135 for 8 mg and P <.001 for 16 mg) or benzoylecgonine concentrations (P =.0277 for 8 mg and P =.006 for 16 mg) during the maintenance phase of the study. For the 16-mg group, mean benzoylecgonine concentrations fell from 3715 ng/mL during baseline to 186 ng/mL during the withdrawal phase; mean morphine concentrations fell from 3311 ng/mL during baseline to 263 ng/mL during withdrawal. For the 8-mg group, mean benzoylecgonine concentrations fell from 6761 ng/mL during baseline to 676 ng/mL during withdrawal; mean morphine concentrations fell from 3890 ng/mL during baseline to 661 ng/mL during withdrawal. Qualitative urinalysis showed a similar pattern of results. Subjects receiving the highest dose showed concomitant decreases in both urine morphine and benzoylecgonine concentrations. There were no significant group differences in treatment retention or adverse events. CONCLUSIONS: A sublingual buprenorphine solution at 16 mg daily is well tolerated and effective in reducing concomitant opiate and cocaine use. The therapeutic effect on cocaine use appears independent of that on opiate use. PMID- 14749695 TI - The effect of echinacea (Echinacea purpurea root) on cytochrome P450 activity in vivo. AB - BACKGROUND: Echinacea is a widely available over-the-counter herbal remedy. Tinctures of echinacea have been shown to inhibit cytochrome P450 (CYP) in vitro. The effect of echinacea (Echinacea purpurea root) on CYP activity in vivo was assessed by use of the CYP probe drugs caffeine (CYP1A2), tolbutamide (CYP2C9), dextromethorphan (CYP2D6), and midazolam (hepatic and intestinal CYP3A). METHODS: Twelve healthy subjects (6 men) completed this 2-period, open-label, fixed schedule study. Caffeine, tolbutamide, dextromethorphan, and oral and intravenous midazolam were administered before and after a short course of echinacea (400 mg 4 times a day for 8 days) to determine in vivo CYP activities. RESULTS: Echinacea administration significantly increased the systemic clearance of midazolam by 34%, from 32 +/- 7 L/h to 43 +/- 16 L/h (P =.003; 90% confidence interval [CI], 116%-150%), and significantly reduced the midazolam area under the concentration time curve by 23%, from 127 +/- 36 microg. h/L to 102 +/- 43 microg. h/L (P =.024; 90% CI, 63%-88%). In contrast, the oral clearance of midazolam was not significantly altered (P =.655; 90% CI, 75%-124%), 137 +/- 19 L/h compared with 146 +/- 71 L/h. The oral availability of midazolam after echinacea dosing was significantly increased (P =.028; 90% CI, 108%-153%), from 0.23 +/- 0.06 to 0.33 +/- 0.13. Hepatic availability (0.72 +/- 0.08 versus 0.61 +/- 0.16; P =.006; 90% CI, 73%-90%) and intestinal availability (0.33 +/- 0.11 versus 0.61 +/- 0.38; P =.015; 90% CI, 125%-203%) were significantly altered in opposite directions. Echinacea dosing significantly reduced the oral clearance of caffeine, from 6.6 +/- 3.8 L/h to 4.9 +/- 2.3 L/h (P =.049; 90% CI, 58%-96%). The oral clearance of tolbutamide was reduced by 11%, from 0.81 +/- 0.18 L/h to 0.72 +/- 0.19 L/h, but this change was not considered to be clinically relevant because the 90% CIs were within the 80% to 125% range. The oral clearance of dextromethorphan in 11 CYP2D6 extensive metabolizers was not affected by echinacea dosing (1289 +/- 414 L/h compared with 1281 +/- 483 L/h; P =.732; 90% CI, 89%-108%). CONCLUSIONS: Echinacea (E purpurea root) reduced the oral clearance of substrates of CYP1A2 but not the oral clearance of substrates of CYP2C9 and CYP2D6. Echinacea selectively modulates the catalytic activity of CYP3A at hepatic and intestinal sites. The type of drug interaction observed between echinacea and other CYP3A substrates will be dependent on the relative extraction of drugs at hepatic and intestinal sites. Caution should be used when echinacea is coadministered with drugs dependent on CYP3A or CYP1A2 for their elimination. PMID- 14749696 TI - Pharmacokinetics and pharmacodynamics of pravastatin in pediatric and adolescent cardiac transplant recipients on a regimen of triple immunosuppression. AB - BACKGROUND: In adults, pravastatin reduces the development and progression of transplant vasculopathy, the main long-term risk after cardiac transplantation. The pharmacokinetics of pravastatin is not known in children taking calcineurin inhibitors. Our aim was to determine the single-dose pharmacokinetics and short term safety of pravastatin in children undergoing regular triple-drug immunosuppressive therapy after cardiac transplantation. METHODS: Nineteen pediatric cardiac transplant recipients (aged 4.4 to 18.9 years) receiving triple immunosuppression therapy consisting of methylprednisolone (19 patients), cyclosporine (INN, cyclosporin) (17 patients) or tacrolimus (2 patients), and azathioprine (18 patients) or mycophenolate mofetil (1 patient) ingested a single 10-mg dose of pravastatin, and plasma pravastatin concentrations were measured up to 24 hours. Subsequently, the patients took 10 mg pravastatin orally once daily for 8 weeks. The lipid-lowering effect and the safety of pravastatin therapy were studied. RESULTS: The mean peak plasma concentration (C(max)) of pravastatin was 122.2 +/- 88.2 ng/mL (range, 11.4-305.0 ng/mL), and the area under the plasma concentration-time curve of pravastatin from 0 to 10 hours [AUC(0-10)] was 264.1 +/- 192.4 ng.h/mL (range, 30.8-701.6 ng.h/mL). These C(max) and AUC(0-10) values are nearly 10-fold higher than the corresponding values reported in hypercholesterolemic children in the absence of immunosuppressive therapy. The time of peak concentration (t(max)) of pravastatin was 1.1 +/- 0.4 hours (range, 0.5-2 hours), and the mean elimination half-life (t(1/2)) was 1.2 +/- 0.3 hours (range, 0.7-2.2 hours); these parameters were similar to those in the hypercholesterolemic children. By 8 weeks of treatment, the concentration of serum total cholesterol decreased by 13% (P =.005), low-density lipoprotein cholesterol by 27% (P <.0001), and triglycerides by 6% (not significant, P =.28); the concentration of high-density lipoprotein cholesterol increased by 7% (not significant, P =.30). No clinically significant increases in serum ALT, creatine kinase, or creatinine levels were observed. CONCLUSIONS: The plasma concentrations of pravastatin in pediatric cardiac recipients receiving triple immunosuppressive medication are nearly 10-fold higher than in hypercholesterolemic children after the same pravastatin dose. However, the short term therapy of pravastatin was well tolerated and effective in lowering serum cholesterol levels in cardiac recipients. PMID- 14749697 TI - Monitoring amiodarone's toxicities: recommendations, evidence, and clinical practice. AB - OBJECTIVES: We sought to develop an explicit evidence-based model of medication monitoring and to evaluate monitoring practices and adverse drug events in patients taking amiodarone at one institution. METHODS: We searched MEDLINE (1966 through 2000) for English-language publications providing specific monitoring recommendations for amiodarone. A cross-sectional retrospective chart review of 99 outpatients receiving amiodarone therapy between Jan 1, 2000, and Jan 1, 2001, at a large tertiary-care hospital was performed to assess monitoring practices. Adverse drug events were identified by use of structured implicit reviews. The main outcome measure was the proportion of patients receiving the monitoring recommended in the literature and having amiodarone-related adverse drug events. RESULTS: Forty-three articles were identified that provided specific monitoring recommendations, although no studies were found that compared the outcomes of patients managed with different monitoring regimens. Overall, 70% of the recommended monitoring criteria were satisfied, although only 9 patients (9%; 95% confidence interval [95% CI], 3%-15%) received all of the recommended monitoring. Variability in monitoring practices was identified at all stages of the monitoring model. Of the patients, 52 (52%; 95% CI, 42%-62%) received minimum baseline evaluations, 22 (22%; 95% CI, 14%-31%) underwent ongoing surveillance, 75 (75%; 95% CI, 61%-89%) had appropriate responses to abnormal surveillance results, and 71 (71%; 95% CI, 62%-80%) had timely follow-up visits. Of the patients, 8 (8%; 95% CI, 3%-13%) had 9 amiodarone-related adverse drug events, of which 3 were judged to be preventable. Interrater agreement for monitoring processes (kappa = 0.83) and adverse drug events (kappa = 0.67) was good. CONCLUSIONS: Current standards for amiodarone toxicity monitoring are based on expert opinion with limited evidence to support most recommendations. Monitoring practices appear to vary significantly, with few patients receiving all of the recommended monitoring. Some amiodarone-related adverse drug events may be preventable and patient safety might be improved with a better understanding of monitoring processes. PMID- 14749698 TI - Do data presented at an American Society for Clinical Pharmacology and Therapeutics meeting make it to full publication? PMID- 14749699 TI - Cellular immune reconstitution after subcutaneous alemtuzumab (anti-CD52 monoclonal antibody, CAMPATH-1H) treatment as first-line therapy for B-cell chronic lymphocytic leukaemia. AB - Little information is available on long-term immune reconstitution after therapy with alemtuzumab in B-CLL patients. We present long-term follow-up data for blood lymphocyte subsets analysed by flow cytometry in previously untreated B-CLL patients who received alemtuzumab subcutaneously as first-line therapy. All lymphoid subsets were significantly (P<0.001) and profoundly reduced; the median end-of-treatment counts for CD4(+), CD8(+), CD3(-)56(+) (natural killer (NK)), CD3(+)56(+) (NK-T) and CD19(+)5(-) (normal B) cells were 43, 20, 4, 1 and 8 cells/microl, respectively. The median cell count of all subsets remained at <25% of the baseline values for >9 months post-treatment. CD4(+) and CD8(+) levels in blood had reached >100 cells/microl in >50% of the patients at 4 months after the end of treatment. One patient had a cytomegalovirus reactivation and one patient developed Pneumocystis carinii pneumonia during therapy. No opportunistic or other major infections were recorded during unmaintained, long-term follow-up. There was no correlation between the cumulative dose of alemtuzumab and the severity or length of immunosuppression. CD52(-) T-cell subsets occurred during the treatment and comprised >80% of all CD4(+) and CD8(+) cells in the blood at the end of therapy. These subpopulations declined gradually during unmaintained follow-up. The relationship between these observations and the safety/antitumour effects of alemtuzumab is discussed. PMID- 14749700 TI - TEL-Syk fusion constitutively activates PI3-K/Akt, MAPK and JAK2-independent STAT5 signal pathways. AB - We previously reported the fusion of the TEL gene to the Syk gene in myelodysplastic syndrome with t(9;12)(q22;p12). TEL-Syk fusion transformed interleukin-3 (IL-3)-dependent murine hematopoietic cell line BaF3 to growth factor independence. Here, we investigate the intracellular signal transduction of the stable transfectants. TEL-Syk fusion protein was associated with the p85 subunit of phosphatidyl inositol 3 kinase (PI3-K) followed by the activation of Akt in the absence of IL-3. Vav, phospholipase C-gamma2 and mitogen-activated protein kinase (MAPK) were also constitutively activated. TEL-Syk also activated the signal transducer and activator of transcription 5 (STAT5) in the absence of Janus kinase 2 activation. None of these kinases were phosphorylated in the BaF3 cells transfected with TELDeltaPNT-Syk in which the oligomerization domain of TEL was deleted. Inhibitor analysis showed that the MAPK pathway was important in TEL Syk-mediated cell proliferation. The immunofluorescence technique revealed that the TEL-Syk fusion protein was located in the cytoplasm. These data suggest that TEL-Syk fusion protein in the cytoplasm leads to the constitutive activation of PI3-K/Akt, MAPK and STAT5 signal pathways, which are closely involved in IL-3 independent cell proliferation of BaF3 cells. PMID- 14749701 TI - Murine marrow cellularity and the concept of stem cell competition: geographic and quantitative determinants in stem cell biology. AB - In unperturbed mice, the marrow cell numbers correlate with the stem cell numbers. High levels of long-term marrow engraftment are obtained with infusion of high levels of marrow cells in untreated mice. To address the issue of stem cell competition vs 'opening space', knowledge of total murine marrow cellularity and distribution of stem and progenitor cells are necessary. We determined these parameters in different mouse strains. Total cellularity in BALB/c mice was 530+/ 20 million cells; stable from 8 weeks to 1 year of age. C57BL/6J mice had 466+/ 48 million marrow cells. Using these data, theoretical models of infused marrow (40 million cells) replacing or adding to host marrow give chimerism values of 7.5 and 7.0%, respectively; the observed 8-week engraftment of 40 million male BALB/c marrow cells into female hosts (72 mice) gave a value of 6.91+/-0.4%. This indicates that syngeneic engraftment is determined by stem cell competition. Our studies demonstrate that most marrow cells, progenitors and engraftable stem cells are in the spine. There was increased concentration of progenitors in the spine. Total marrow harvest for stem cell purification and other experimental purposes was both mouse and cost efficient with over a four-fold decrease in animal use and a financial saving. PMID- 14749702 TI - Second malignancy after treatment of adult acute myeloid leukemia: cohort study on adult patients enrolled in the GIMEMA trials. PMID- 14749703 TI - Deletion and reduced expression of the Fanconi anemia FANCA gene in sporadic acute myeloid leukemia. AB - Fanconi anemia (FA) is an autosomal recessive chromosomal instability disorder caused by mutations in one of seven known genes (FANCA,C,D2,E,F,G and BRCA2). Mutations in the FANCA gene are the most prevalent, accounting for two-thirds of FA cases. Affected individuals have greatly increased risks of acute myeloid leukemia (AML). This raises the question as to whether inherited or acquired mutations in FA genes might be involved in the development of sporadic AML. Quantitative fluorescent PCR was used to screen archival DNA from sporadic AML cases for FANCA deletions, which account for 40% of FANCA mutations in FA homozygotes. Four heterozygous deletions were found in 101 samples screened, which is 35-fold higher than the expected population frequency for germline FANCA deletions (P<0.0001). Sequencing FANCA in the AML samples with FANCA deletions did not detect mutations in the second allele and there was no evidence of epigenetic silencing by hypermethylation. However, real-time quantitative PCR analysis in these samples showed reduced expression of FANCA compared to nondeleted AML samples and to controls. These findings suggest that gene deletions and reduced expression of FANCA may be involved in the promotion of genetic instability in a subset of cases of sporadic AML. PMID- 14749704 TI - Inhibition of survivin expression suppresses the growth of aggressive non Hodgkin's lymphoma. AB - Survivin is a member of the inhibitor of apoptosis protein (IAP) family and functions both as an apoptosis inhibitor and a regulator of cell division. Survivin overexpression is common in many human tumors and correlates with survival in large cell non-Hodgkin's lymphoma. To evaluate this molecule as a potential therapeutic target in large-cell lymphoma, we evaluated the effect of survivin inhibition both in vitro and in vivo. Using an antisense oligonucleotide (ASO) approach, cell growth was significantly inhibited in the DoHH2, RL and HT lymphoma cell lines. In a lymphoma xenograft model, the development of tumors as well as the growth of established tumors was inhibited in the survivin ASO treated mice compared to controls. To assess the efficacy of the survivin ASO in combination with other biological agents, we combined the survivin ASO with an anti-CD20 monoclonal antibody, rituximab. The effect of survivin ASO and rituximab in combination was additive in vitro. In vivo, however, suppression of tumor growth with the combination was not significantly superior to controls. We conclude that inhibition of survivin expression is an attractive therapeutic strategy in aggressive non-Hodgkin's lymphomas, and that combining survivin ASO with rituximab may enhance the efficacy of this approach. PMID- 14749705 TI - CD38 expression on B-cell chronic lymphocytic leukemic cells is strongly correlated with vascular endothelial growth factor expression. PMID- 14749706 TI - Retinoic acid syndrome in NOD/scid mice induced by injecting an acute promyelocytic leukemia cell line. AB - All-trans retinoic acid (ATRA) induces complete remission in patients with acute promyelocytic leukemia (APL). However, ATRA sometimes causes retinoic acid syndrome (RAS) characterized by respiratory distress, pleural effusions, fever and weight gain. To investigate the pathophysiology of RAS, we generated an animal model by injecting an APL cell line, NB4, into immunodeficient mice. When NOD/scid mice were injected intravenously with fully differentiated NB4 cells (1 x 10(7)) and then given a daily administration of ATRA, three of 12 mice died of pulmonary edema within 14 days. Pathologically, dilated lung capillary vessels and alveolar effusions were observed. After the injection, NB4 cells were detected in the lung within 2 days and in the pleural effusion later on. The gene expression levels of CXC chemokines (MIP-2 and KC) and ICAM-1 were increased in the lung and heart by the ATRA administration. In immunohistochemical analyses, MIP-2 was clearly detected in alveolar macrophages of the lung in mice with RAS. Dexamethasone treatment prevented the development of RAS and decreased the CXC chemokine mRNA expression in the lung. These findings suggested that the activation of adhesion molecules for leukocytes and expression of CXC chemokines in the lung are closely involved in triggering RAS. PMID- 14749707 TI - Effect of thalidomide therapy on bone marrow angiogenesis in multiple myeloma. AB - Bone marrow (BM) angiogenesis is increased in multiple myeloma and is an important prognostic factor for survival. Previous studies have shown that BM angiogenesis does not change following chemotherapy or stem cell transplant. Given its potential antiangiogenic effect, we evaluated if thalidomide therapy would affect the BM microvessel density (MVD). We studied BM angiogenesis in 81 patients with various disease stages treated with thalidomide with or without dexamethasone. MVD was determined as previously described. MVD was compared between pretreatment marrows and those obtained 4-6 months following therapy. The median (range) MVD pretherapy was 28 (2-116) and post-therapy was 15 (3-97). A partial or complete response was seen in 58% of patients, stable disease in 41% and progressive disease in one patient. MVD decreased significantly in responders (median decrease of 12, P<0.001). In contrast, no significant change in MVD was seen in those failing to respond to thalidomide. Unlike the lack of resolution of angiogenesis reported with other therapies, we demonstrate for the first time a significant decrease in microvessels with thalidomide therapy. Although not conclusive, this result lends further support to the hypothesis that angiogenesis is a relevant therapeutic target in myeloma. PMID- 14749708 TI - BRAF gene is not mutated in mismatch repair-proficient or -deficient plasma cell dyscrasias. PMID- 14749709 TI - Idiopathic congenital nonobstructive nephrolithiasis: a case report and review. AB - We describe a case of congenital nephrolithiasis, which presented with hematuria at birth. No etiopathological factor could be determined for renal stone formation despite extensive investigation. There was a family history of renal stones in both maternal and paternal grandparents and of microscopic hematuria in the mother. There was no associated urinary flow obstruction and the hematuria remitted spontaneously. The infant was treated conservatively. The follow-up studies revealed persistence of renal stone but no hematuria. This case can be considered as idiopathic, or an early presentation of one of the rare genetic disorders associated with renal stones. Congenital nonobstructive nephrolithiasis has not been reported previously and should be considered as a cause of hematuria during this age. PMID- 14749710 TI - Fetal/neonatal supraventricular tachycardia. AB - At 38 weeks of gestation, a woman with a previously unremarkable pregnancy was noted to have fetal tachycardia without obvious cause. Fetal echocardiography resulted in a presumptive diagnosis of fetal atrial flutter with a 2:1 block. The newborn resumed the same rhythm. The neonate underwent transesophageal incremental overdrive pacing. A normal sinus rhythm was restored. The infant had no recurrence to age 6 months. PMID- 14749711 TI - Association between pulse pressure, carotid intima-media thickness and carotid and/or iliofemoral plaque in hypertensive patients. AB - Increased common carotid artery intima-media thickness (CCA-IMT) and carotid and/or iliofemoral (C/IF) plaque are frequent in subjects treated for hypertension, but their association with pulse pressure (PP) has rarely been studied. Using ultrasound techniques, CCA-IMT and C/IF plaques were studied in 323 hypertensive subjects, who were classified into four groups according to the adequacy of blood pressure (BP) control (systolic BP (SBP) <140 mmHg and diastolic BP (DBP) <90 mmHg) and PP (high or low). After adjustment for confounding variables, an increase in CCA-IMT was the only factor significantly and independently associated with high PP, irrespective of the effectiveness of blood pressure control and of antihypertensive drug treatment. CCA-IMT correlated with age, PP, waist-to-hip ratio, tobacco consumption, and heart rate. C/IF plaques correlated with age, tobacco consumption, diabetes mellitus, and dyslipidaemia. To conclude, even with SBP<140 mmHg and DBP<90 mmHg on treatment, hypertensive subjects may have increased CCA-IMT values and C/IF plaque. Four cardiovascular risk factors seem to be involved in these alterations, namely tobacco consumption, dyslipidaemia, diabetes and increased PP. Only the latter factor does not have a standardized effective treatment. PMID- 14749712 TI - The high-risk and population strategy for occupational health promotion (HIPOP OHP) study: study design and cardiovascular risk factors at the baseline survey. AB - In order to establish the methodology of a population strategy for improving cardiovascular risk factors, we have planned the High-risk and Population Strategy for Occupational Health Promotion Study (HIPOP-OHP study). This study is a nonrandomized control trial in approximately 6500 participants in six intervention and six control companies. Our population strategy is based on three factors, nutrition, physical activity, and smoking. For each factor, a researcher's working team was organized and has been supporting the intervention. A standardized method to obtain comparable data has also been established. In the baseline survey, urinary sodium excretion in male subjects was higher, and urinary potassium excretion was lower in both genders in the intervention group compared to the control group. The prevalence of hypertension for both genders was also higher in the intervention group. Male subjects in the intervention group had higher serum total cholesterol than controls, while high-density lipoprotein cholesterol was lower in both genders in the intervention group compared to the control group. These differences were reflected by our finding that the predicted relative risk of coronary heart disease for male subjects was significantly higher in the intervention group (relative risk, RR: 1.17; 95% confidence interval, 95% CI.: 1.09, 1.25) and significantly lower in the control group (RR: 0.93; 95% CI.: 0.89, 0.98) compared to a model Japanese population. Similar results were observed in the female subjects. Taken together, these findings indicate that it is possible to compare trends of predicted relative risk for coronary heart disease between two groups. PMID- 14749713 TI - Reproducibility of nocturnal blood pressure fall in early phases of untreated essential hypertension: a prospective observational study. AB - A number of studies have shown that a smaller than normal nocturnal blood pressure (BP) decrease is associated with cardiovascular disease. However, no large prospective studies have examined the reliability of nocturnal dipping within individuals. The aim of our study was to investigate the short-term variability of nocturnal BP fall in a large cohort of patients with recently diagnosed essential hypertension. In all, 414 uncomplicated never treated hypertensive patients referred to our outpatient hypertension hospital clinic (mean age 46+/-12 years; 257 M, 157 F) prospectively underwent: (1). repeated clinic BP measurements; (2). routine examinations recommended by WHO/ISH guidelines; and (3). ambulatory BP monitoring (ABPM) twice within a 4-week period. Dipping pattern was defined as a reduction in the average systolic and diastolic BP at night greater than 10% compared to average daytime values. Overall, 311 patients (75.1%) showed no change in their diurnal variations in BP. Of the 278 patients who had a dipping pattern on the first ABPM, 219 (78.7%) confirmed this type of profile on the second ABPM, while 59 (21.3%) showed a nondipping pattern. Among 37 dipper patients with >20% of nocturnal systolic BP decrease (extreme dippers), only 16 (43.2%) had this marked fall in BP on the second ABPM. Of the 136 patients who had a nondipping pattern on the first ABPM, 92 (67.6%) confirmed their initial profile on the second ABPM, while 44 (32.4%) did not. Patients with reproducible nondipping profile were older (48+/-12 years) than those with reproducible dipping profile (44+/-12 years, P<0.05). These findings indicate that: (1). short-term reproducibility of nocturnal fall in BP in untreated middle-aged hypertensives is rather limited: overall, one-fourth of patients changed their initial dipping patterns when they were studied again after a few weeks; (2). this was particularly true for extreme dipping and nondipping patterns; (3). abnormalities in nocturnal BP fall, assessed by a single ABPM, cannot be taken as independent predictors of increased cardiovascular risk. PMID- 14749714 TI - A review of the MHC genetics of rheumatoid arthritis. AB - Rheumatoid arthritis is a common complex genetic disease, and, despite a significant genetic element, no gene other than HLA-DRB1 has been clearly demonstrated to be involved in the disease. However, this association accounts for less than half the overall genetic susceptibility. Investigation of other candidate genes, in particular those that reside within the major histocompatibility complex, are hampered by the presence of strong linkage disequilibrium and problems with study design. PMID- 14749715 TI - Inhibition of synapse assembly in mammalian muscle in vivo by RNA interference. AB - The formation of the vertebrate neuromuscular junction (NMJ) requires the receptor tyrosine kinase MuSK and the adaptor molecule rapsyn. Here, we report that the phenotypes of mice deficient in these two molecules can be reproduced by RNA interference (RNAi) in rat muscle in vivo. Specifically, double-stranded RNA (dsRNA) targeting MuSK and rapsyn inhibited the formation of the NMJ in rat muscle fibres in vivo, while dsRNA targeting nonessential proteins did not have any effect. Moreover, plasmids that trigger RNAi to MuSK induced the disassembly of existing NMJs. These results thus demonstrate for the first time the functionality of dsRNA in silencing endogenous genes in adult mammalian muscle in vivo. Moreover, they show that MuSK is also required for the maintenance of the NMJ, offering a mechanistic explanation for the myasthenia gravis caused by auto antibodies to MuSK. PMID- 14749716 TI - Characterization of the interactions between mammalian PAZ PIWI domain proteins and Dicer. AB - PAZ PIWI domain (PPD) proteins, together with the RNA cleavage products of Dicer, form ribonucleoprotein complexes called RNA-induced silencing complexes (RISCs). RISCs mediate gene silencing through targeted messenger RNA cleavage and translational suppression. The PAZ domains of PPD and Dicer proteins were originally thought to mediate binding between PPD proteins and Dicer, although no evidence exists to support this theory. Here we show that PAZ domains are not required for PPD protein-Dicer interactions. Rather, a subregion of the PIWI domain in PPD proteins, the PIWI-box, binds directly to the Dicer RNase III domain. Stable binding between PPD proteins and Dicer was dependent on the activity of Hsp90. Unexpectedly, binding of PPD proteins to Dicer inhibits the RNase activity of this enzyme in vitro. Lastly, we show that PPD proteins and Dicer are present in soluble and membrane-associated fractions, indicating that interactions between these two types of proteins may occur in multiple compartments. PMID- 14749717 TI - Involvement of ASK1 in Ca2+-induced p38 MAP kinase activation. AB - The mammalian mitogen-activated protein (MAP) kinase kinase kinase apoptosis signal-regulating kinase 1 (ASK1) is a pivotal component in cytokine- and stress induced apoptosis. It also regulates cell differentiation and survival through p38 MAP kinase activation. Here we show that Ca2+ signalling regulates the ASK1 p38 MAP kinase cascade. Ca2+ influx evoked by membrane depolarization in primary neurons and synaptosomes induced activation of p38, which was impaired in those derived from ASK1-deficient mice. Ca2+/calmodulin-dependent protein kinase type II (CaMKII) activated ASK1 by phosphorylation. Moreover, p38 activation induced by the expression of constitutively active CaMKII required endogenous ASK1. Thus, ASK1 is a critical intermediate of Ca2+ signalling between CaMKII and p38 MAP kinase. PMID- 14749718 TI - Size matters. Workshop on growth control in development and disease. PMID- 14749719 TI - p41-Arc subunit of human Arp2/3 complex is a p21-activated kinase-1-interacting substrate. AB - The formation of new branched actin filament networks at the cell cortex of migrating cells is choreographed by the actin-related protein (Arp) 2/3 complex. Despite the fundamental role of the Arp2/3 complex in actin nucleation and branching, upstream signals that control the functions of p41-Arc, a putative regulatory component of the mammalian Arp2/3 complex, remain unidentified. Here we show that p41-Arc interacts with p21-activated kinase 1 (Pak1) both in vitro and in vivo. Pak1 phosphorylation of p41-Arc regulates its localization with the Arp2/3 complex in the cortical nucleation regions of cells. Pak1 phosphorylates p41-Arc on threonine 21 in the first WD repeat, and its mutation has functional implications in vivo. Threonine 21 phosphorylation by Pak1 is required for both constitutive and growth-factor-induced cell motility. Pak1 regulation of p41-Arc activation status represents a novel mechanism by which signalling pathways may influence the functions of the Arp2/3 complex, leading to motility in mammalian cells. PMID- 14749720 TI - Analysis of Scc1-deficient cells defines a key metaphase role of vertebrate cohesin in linking sister kinetochores. AB - Cleavage of the cohesin subunit Scc1p/Mcd1p/Rad21 permits sister chromatid separation and is considered to trigger anaphase onset. It has also been suggested that the cohesin complex is essential for chromosome condensation and for assembling fully functional kinetochores. Here, we used vertebrate cells conditionally deficient in Scc1 to probe cohesin function in mitosis. Cells lacking cohesin arrest in prometaphase, with many chromosomes failing to align at a metaphase plate and high levels of the spindle assembly checkpoint protein, BubR1, at all kinetochores. We show that the structural integrity of chromosomes is normal in the absence of Scc1. Furthermore, specific inhibition of topoisomerase II, which is required for decatenation of replicated chromosomes, can bypass the cohesin requirement for metaphase chromosome alignment and spindle checkpoint silencing. Since the kinetochore effects of Scc1 deficiency can be compensated for by topoisomerase II inhibition, we conclude that Scc1 is not absolutely required for kinetochore assembly or function, and that its principal role in allowing the onset of anaphase is the establishment of sufficient inter sister tension to allow biorientation. PMID- 14749721 TI - Toll-like receptors differentially induce nucleosome remodelling at the IL-12p40 promoter. AB - Toll-like receptors (TLRs) mediate recognition of microbial components. Despite activation of a shared set of signal transduction molecules, the biological effects of certain TLR agonists differ considerably. In macrophages and dendritic cells, stimulation by the prototypical stimuli CpG-DNA (TLR9), lipopolysaccharide (LPS; TLR4) and lipoteichoic acid (LTA; TLR2) resulted in striking differences in expression of IL-12. However, these stimuli induced similar amounts of the common proinflammatory cytokine TNFalpha. Surprisingly, an IL-12p40 promoter reporter construct was activated equally by CpG-DNA, LPS and LTA. Examinations of the chromatin structure of the endogenous IL-12p40 promoter revealed that nucleosome remodelling contributed to differential IL-12 induction. Upon stimulation, nucleosome architecture was changed to provide increased access to the IL-12p40 promoter. In dendritic cells, a differential induction of nucleosome remodelling at the IL-12p40 promoter was observed upon triggering with different TLR agonists. These results identify nucleosome remodelling as an additional restriction point in differential TLR signalling. PMID- 14749722 TI - Parasite-specific immune response in adult Drosophila melanogaster: a genomic study. AB - Insects of the order Diptera are vectors for parasitic diseases such as malaria, sleeping sickness and leishmania. In the search for genes encoding proteins involved in the antiparasitic response, we have used the protozoan parasite Octosporea muscaedomesticae for oral infections of adult Drosophila melanogaster. To identify parasite-specific response molecules, other flies were exposed to virus, bacteria or fungi in parallel. Analysis of gene expression patterns after 24 h of microbial challenge, using Affymetrix oligonucleotide microarrays, revealed a high degree of microbe specificity. Many serine proteases, key intermediates in the induction of insect immune responses, were uniquely expressed following infection of the different organisms. Several lysozyme genes were induced in response to Octosporea infection, while in other treatments they were not induced or downregulated. This suggests that lysozymes are important in antiparasitic defence. PMID- 14749723 TI - DJ-1 has a role in antioxidative stress to prevent cell death. AB - Deletion and point (L166P) mutations of DJ-1 have recently been shown to be responsible for the onset of familial Parkinson's disease (PD, PARK7). The aim of this study was to determine the role of DJ-1 in PD. We first found that DJ-1 eliminated hydrogen peroxide in vitro by oxidizing itself. We then found that DJ 1 knockdown by short interfering RNA rendered SH-SY5Y neuroblastoma cells susceptible to hydrogen peroxide-, MPP+- or 6-hydroxydopamine-induced cell death and that cells harbouring mutant forms of DJ-1, including L166P, became susceptible to death in parallel with the loss of oxidized forms of DJ-1. These results clearly showed that DJ-1 has a role in the antioxidative stress reaction and that mutations of DJ-1 lead to cell death, which is observed in PD. PMID- 14749724 TI - Take five--BACE and the gamma-secretase quartet conduct Alzheimer's amyloid beta peptide generation. AB - In 1959, Dave Brubeck and Paul Desmond revolutionized modern jazz music by composing their unforgettable Take Five in 5/4, one of the most defiant time signatures in all music. Of similar revolutionary importance for biomedical and basic biochemical research is the identification of the minimal set of genes required to obtain a deadly time bomb ticking in all of us: Alzheimer's disease. It now appears that one needs to Take Five genes to produce a deadly peptide by a proteolytic mechanism, which paradoxically is otherwise of pivotal importance for development and cell fate decisions. PMID- 14749725 TI - Endogenous TGF-beta signaling suppresses maturation of osteoblastic mesenchymal cells. AB - Transforming growth factor-beta (TGF-beta), one of the most abundant cytokines in bone matrix, has positive and negative effects on bone formation, although the molecular mechanisms of these effects are not fully understood. Bone morphogenetic proteins (BMPs), members of the TGF-beta superfamily, induce bone formation in vitro and in vivo. Here, we show that osteoblastic differentiation of mouse C2C12 cells was greatly enhanced by the TGF-beta type I receptor kinase inhibitor SB431542. Endogenous TGF-beta was found to be highly active, and induced expression of inhibitory Smads during the maturation phase of osteoblastic differentiation induced by BMP-4. SB431542 suppressed endogenous TGF beta signaling and repressed the expression of inhibitory Smads during this period, possibly leading to acceleration of BMP signaling. SB431542 also induced the production of alkaline phosphatase and bone sialoprotein, and matrix mineralization of human mesenchymal stem cells. Thus, signaling cross-talk between BMP and TGF-beta pathways plays a crucial role in the regulation of osteoblastic differentiation, and TGF-beta inhibitors may be invaluable for the treatment of various bone diseases by accelerating BMP-induced osteogenesis. PMID- 14749726 TI - A protein kinase target of a PDK1 signalling pathway is involved in root hair growth in Arabidopsis. AB - Here we report on a lipid-signalling pathway in plants that is downstream of phosphatidic acid and involves the Arabidopsis protein kinase, AGC2-1, regulated by the 3'-phosphoinositide-dependent kinase-1 (AtPDK1). AGC2-1 specifically interacts with AtPDK1 through a conserved C-terminal hydrophobic motif that leads to its phosphorylation and activation, whereas inhibition of AtPDK1 expression by RNA interference abolishes AGC2-1 activity. Phosphatidic acid specifically binds to AtPDK1 and stimulates AGC2-1 in an AtPDK1-dependent manner. AtPDK1 is ubiquitously expressed in all plant tissues, whereas expression of AGC2-1 is abundant in fast-growing organs and dividing cells, and activated during re-entry of cells into the cell cycle after sugar starvation-induced G1-phase arrest. Plant hormones, auxin and cytokinin, synergistically activate the AtPDK1 regulated AGC2-1 kinase, indicative of a role in growth and cell division. Cellular localisation of GFP-AGC2-1 fusion protein is highly dynamic in root hairs and at some stages confined to root hair tips and to nuclei. The agc2-1 knockout mutation results in a reduction of root hair length, suggesting a role for AGC2-1 in root hair growth and development. PMID- 14749727 TI - Human Fip1 is a subunit of CPSF that binds to U-rich RNA elements and stimulates poly(A) polymerase. AB - In mammals, polyadenylation of mRNA precursors (pre-mRNAs) by poly(A) polymerase (PAP) depends on cleavage and polyadenylation specificity factor (CPSF). CPSF is a multisubunit complex that binds to the canonical AAUAAA hexamer and to U-rich upstream sequence elements on the pre-mRNA, thereby stimulating the otherwise weakly active and nonspecific polymerase to elongate efficiently RNAs containing a poly(A) signal. Based on sequence similarity to the Saccharomyces cerevisiae polyadenylation factor Fip1p, we have identified human Fip1 (hFip1) and found that the protein is an integral subunit of CPSF. hFip1 interacts with PAP and has an arginine-rich RNA-binding motif that preferentially binds to U-rich sequence elements on the pre-mRNA. Recombinant hFip1 is sufficient to stimulate the in vitro polyadenylation activity of PAP in a U-rich element-dependent manner. hFip1, CPSF160 and PAP form a ternary complex in vitro, suggesting that hFip1 and CPSF160 act together in poly(A) site recognition and in cooperative recruitment of PAP to the RNA. These results show that hFip1 significantly contributes to CPSF-mediated stimulation of PAP activity. PMID- 14749728 TI - The region 3' to Xist mediates X chromosome counting and H3 Lys-4 dimethylation within the Xist gene. AB - A counting process senses the X chromosome/autosome ratio and ensures that X chromosome inactivation (XCI) initiates in the female (XX) but not in the male (XY) mouse embryo. Counting is regulated by the X-inactivation centre, which contains the Xist gene. Deleting 65 kb 3' to Xist in XO embryonic stem (ES) cells affects counting and results in inappropriate XCI upon differentiation. We show here that normal counting can be rescued in these deleted ES cells using cre/loxP re-insertion, and refine the location of elements controlling counting within a 20 kb bipartite domain. Furthermore, we show that the 65 kb deletion also leads to inappropriate XCI in XY differentiated ES cells, which excludes the involvement of sex-specific mechanisms in the initiation of XCI. At the chromatin level, we have found that the Xist gene corresponds to a peak of H3 Lys-4 dimethylation, which is dramatically and specifically affected by the deletion 3' to Xist. Our results raise the possibility that H3 Lys-4 dimethylation within Xist may be functionally implicated in the counting process. PMID- 14749729 TI - Regulated exocytosis of an H+/myo-inositol symporter at synapses and growth cones. AB - Phosphoinositides, synthesized from myo-inositol, play a critical role in the development of growth cones and in synaptic activity. As neurons cannot synthesize inositol, they take it up from the extracellular milieu. Here, we demonstrate that, in brain and PC12 cells, the recently identified H(+)/myo inositol symporter HMIT is present in intracellular vesicles that are distinct from synaptic and dense-core vesicles. We further show that HMIT can be triggered to appear on the cell surface following cell depolarization, activation of protein kinase C or increased intracellular calcium concentrations. HMIT cell surface expression takes place preferentially in regions of nerve growth and at varicosities and leads to increased myo-inositol uptake. The symporter is then endocytosed in a dynamin-dependent manner and becomes available for a subsequent cycle of stimulated exocytosis. HMIT is thus expressed in a vesicular compartment involved in activity-dependent regulation of myo-inositol uptake in neurons. This may be essential for sustained signaling and vesicular traffic activities in growth cones and at synapses. PMID- 14749730 TI - Differential functional interplay of TOGp/XMAP215 and the KinI kinesin MCAK during interphase and mitosis. AB - XMAP215/TOGp family members and KinI kinesins are conserved microtubule (MT) regulatory proteins, and have been viewed as possessing prominent antagonistic stabilizing/destabilizing activities that must be balanced. Here, interdependencies between TOGp and the KinI kinesin MCAK were analyzed in human leukemia cells. A system was established that permits inducible overexpression in homogeneous cell populations that simultaneously synthesize interfering short hairpin RNAs. We present evidence that the functional interplay of TOGp and MCAK proteins is manifested as three distinct phenotypes during the cell cycle. The first involves a role for TOGp in protecting spindle MTs from MCAK activity at the centrosome, which appears essential to prevent the formation of disorganized multipolar spindles. The second phenotype involves TOGp-dependent counteraction of excessive MCAK activity during mitosis, which recapitulates the previously established plus-end specific counteractive activities in vitro. The third involves an unexpected destabilization of the interphase MTs by overexpressed TOGp, a phenotype that requires endogenous MCAK. We hypothesize that TOGp dependent prevention of MCAK-mediated spindle disorganization, as evidenced by depletion experiments, reflects a primary physiological role for TOGp in human somatic cells. PMID- 14749731 TI - The protein kinase PKR: a molecular clock that sequentially activates survival and death programs. AB - Cell death and survival play a key role in the immune system as well as during development. The control mechanisms that balance cell survival against cell death are not well understood. Here we report a novel strategy used by a single protein to regulate chronologically cell survival and death. The interferon-induced protein kinase PKR acts as a molecular clock by using catalysis-dependent and independent activities to temporally induce cell survival prior to cell death. We show that the proapoptotic protein PKR surprisingly activates a survival pathway, which is mediated by NF-kappaB to delay apoptosis. Cell death is then induced by PKR through the phosphorylation of eIF-2alpha. This unique temporal control might serve as a paradigm for other kinases whose catalytic activity is not required for all of their functions. PMID- 14749732 TI - Hsp42 is the general small heat shock protein in the cytosol of Saccharomyces cerevisiae. AB - Small heat shock proteins (sHsps) are ubiquitous molecular chaperones that prevent the unspecific aggregation of proteins. So far, Hsp26 was the only unambiguously identified member of the sHsp family in Saccharomyces cerevisiae. We show here that the sHsp system in the cytosol of S. cerevisiae consists of two proteins, Hsp26 and Hsp42. Hsp42 forms large dynamic oligomers with a barrel-like structure. In contrast to Hsp26, which functions predominantly at heat shock temperatures, Hsp42 is active as a chaperone under all conditions tested in vivo and in vitro. Under heat shock conditions, both Hsp42 and Hsp26 suppress the aggregation of one-third of the cytosolic proteins. This subset is about 90% overlapping for Hsp42 and Hsp26. The sHsp substrates belong to different biochemical pathways. This indicates a general protective function of sHsps for proteome stability in S. cerevisiae. Consistent with this observation, sHsp knockout strains show phenotypical defects. Taken together, our results define Hsp42 as an important player for protein homeostasis at physiological and under stress conditions. PMID- 14749733 TI - Molecular clearance of ataxin-3 is regulated by a mammalian E4. AB - Insoluble aggregates of polyglutamine-containing proteins are usually conjugated with ubiquitin in neurons of individuals with polyglutamine diseases. We now show that ataxin-3, in which the abnormal expansion of a polyglutamine tract is responsible for spinocerebellar ataxia type 3 (SCA3), undergoes ubiquitylation and degradation by the proteasome. Mammalian E4B (UFD2a), a ubiquitin chain assembly factor (E4), copurified with the polyubiquitylation activity for ataxin 3. E4B interacted with, and thereby mediated polyubiquitylation of, ataxin-3. Expression of E4B promoted degradation of a pathological form of ataxin-3. In contrast, a dominant-negative mutant of E4B inhibited degradation of this form of ataxin-3, resulting in the formation of intracellular aggregates. In a Drosophila model of SCA3, expression of E4B suppressed the neurodegeneration induced by an ataxin-3 mutant. These observations suggest that E4 is a rate-limiting factor in the degradation of pathological forms of ataxin-3, and that targeted expression of E4B is a potential gene therapy for SCA3. PMID- 14749734 TI - Improved glucose homeostasis and enhanced insulin signalling in Grb14-deficient mice. AB - Gene targeting was used to characterize the physiological role of growth factor receptor-bound (Grb)14, an adapter-type signalling protein that associates with the insulin receptor (IR). Adult male Grb14(-/-) mice displayed improved glucose tolerance, lower circulating insulin levels, and increased incorporation of glucose into glycogen in the liver and skeletal muscle. In ex vivo studies, insulin-induced 2-deoxyglucose uptake was enhanced in soleus muscle, but not in epididymal adipose tissue. These metabolic effects correlated with tissue specific alterations in insulin signalling. In the liver, despite lower IR autophosphorylation, enhanced insulin-induced tyrosine phosphorylation of insulin receptor substrate (IRS)-1 and activation of protein kinase B (PKB) was observed. In skeletal muscle, IR tyrosine phosphorylation was normal, but signalling via IRS-1 and PKB was increased. Finally, no effect of Grb14 ablation was observed on insulin signalling in white adipose tissue. These findings demonstrate that Grb14 functions in vivo as a tissue-specific modulator of insulin action, most likely via repression of IR-mediated IRS-1 tyrosine phosphorylation, and highlight this protein as a potential target for therapeutic intervention. PMID- 14749735 TI - XRCC3 deficiency results in a defect in recombination and increased endoreduplication in human cells. AB - XRCC3 was inactivated in human cells by gene targeting. Consistent with its role in homologous recombination, XRCC3(-/-) cells showed a two-fold sensitivity to DNA cross-linking agents, a mild reduction in sister chromatid exchange, impaired Rad51 focus formation and elevated chromosome aberrations. Furthermore, endoreduplication was increased five- seven-fold in the mutants. The T241M variant of XRCC3 has been associated with an increased cancer risk. Expression of the wild-type cDNA restored this phenotype, while expression of the variant restored the defective recombinational repair, but not the increased endoreduplication. RPA, a protein essential for homologous recombination and DNA replication, is associated with XRCC3 and Rad52. Overexpression of RPA promoted endoreduplication, which was partially complemented by overexpression of the wild type XRCC3 protein, but not by overexpression of the variant protein. Overexpression of Rad52 prevented endoreduplication in RPA-overexpressing cells, in XRCC3(-/-) cells and in the variant-expressing cells, suggesting that deregulated RPA was responsible for the increased endoreduplication. These observations offer the first genetic evidence for the association between homologous recombination and replication initiation having a role in cancer susceptibility. PMID- 14749736 TI - A glycosylated type I membrane protein becomes cytosolic when peptide: N glycanase is compromised. AB - The human cytomegalovirus-encoded glycoprotein US2 catalyzes proteasomal degradation of Class I major histocompatibility complex (MHC) heavy chains (HCs) through dislocation of the latter from the endoplasmic reticulum (ER) to the cytosol. During this process, the Class I MHC HCs are deglycosylated by an N glycanase-type activity. siRNA molecules designed to inhibit the expression of the light chain, beta(2)-microglobulin, block the dislocation of Class I MHC molecules, which implies that US2-dependent dislocation utilizes correctly folded Class I MHC molecules as a substrate. Here we demonstrate it is peptide: N glycanase (PNGase or PNG1) that deglycosylates dislocated Class I MHC HCs. Reduction of PNGase activity by siRNA expression in US2-expressing cells inhibits deglycosylation of Class I MHC HC molecules. In PNGase siRNA-treated cells, glycosylated HCs appear in the cytosol, providing the first evidence for the presence of an intact N-linked type I membrane glycoprotein in the cytosol. N glycanase activity is therefore not required for dislocation of glycosylated Class I MHC molecules from the ER. PMID- 14749737 TI - Long-chain polyunsaturated fatty acids interact with nitric oxide, superoxide anion, and transforming growth factor-beta to prevent human essential hypertension. AB - Patients with uncontrolled essential hypertension have elevated concentrations of superoxide anion (O(2)(-*)), hydrogen peroxide (H(2)O(2)), lipid peroxides, endothelin, and transforming growth factor-beta (TGF-beta) with a simultaneous decrease in endothelial nitric oxide (eNO), superoxide dismutase (SOD), vitamin E, and long-chain polyunsaturated fatty acids (LCPUFAs). Physiological concentrations of angiotensin II activate NAD(P)H oxidase and trigger free radical generation (especially that of O(2)(-*)). Normally, angiotensin II induced oxidative stress is abrogated by adequate production and release of eNO, which quenches O(2)(-*) to restore normotension. Angiotensin II also stimulates the production of endothelin and TGF-beta. TGF-beta enhances NO generation, which in turn suppresses TGF-beta production. Thus, NO has a regulatory role on TGF beta production and is also a physiological antagonist of endothelin. Antihypertensive drugs suppress the production of O(2)(-*) and TGF-beta and enhance eNO synthesis to bring about their beneficial actions. LCPUFAs suppress angiotensin-converting enzyme (ACE) activity, reduce angiotensin II formation, enhance eNO generation, and suppress TGF-beta expression. Perinatal supplementation of LCPUFAs decreases insulin resistance and prevents the development of hypertension in adult life, whereas deficiency of LCPUFAs in the perinatal period results in raised blood pressure later in life. Patients with essential hypertension have low concentrations of various LCPUFAs in their plasma phospholipid fraction. Based on this, it is proposed that LCPUFAs serve as endogenous regulators of ACE activity, O(2)(-*), eNO generation, and TGF-beta expression. Further, LCPUFAs have actions similar to statins, inhibit (especially omega-3 fatty acids) cyclooxygenase activity and suppress the synthesis of proinflammatory cytokines, and activate the parasympathetic nervous system, all actions that reduce the risk of major vascular events. Hence, it is proposed that availability of adequate amounts of LCPUFAs during the critical periods of growth prevents the development of hypertension in adulthood. PMID- 14749738 TI - Cigarette smoking during pregnancy in rural Nepal. Risk factors and effects of beta-carotene and vitamin A supplementation. AB - OBJECTIVE: We examined risk factors of smoking and the association between smoking and pregnancy-related and 6-month infant mortality in rural Nepal, where 30% women reported smoking during pregnancy. DESIGN: Cross-sectional analysis of risk factors associated with smoking status and health consequences of smoking, using prospective data collected as part of a randomized community trial to examine the effect of maternal vitamin A or beta-carotene supplementation on maternal mortality. SETTING: Rural, southeastern plains of Nepal. SUBJECTS AND METHODS: A total of 17 767 women contributed at least one pregnancy during 3.5 y of the study. Data on cigarette or bidi (rolled tobacco) smoking were collected using a 7-day recall, twice during pregnancy. Associations between smoking status and maternal diet, morbidity profile, household socioeconomic status and serum concentration of retinol, carotenoids and tocopherols were examined. Further, relative risk (RR) and 95% confidence intervals (CI) were calculated to estimate supplement effects on pregnancy-related mortality, stratified by smoking status during pregnancy. RESULTS: Smokers were more likely to be older, illiterate and poor compared to nonsmokers. Fruit and vegetable consumption among smokers and nonsmokers did not vary. However, smokers were more likely to consume meat/fish/eggs and less likely to consume milk than nonsmokers. They were also more likely to report symptoms of vaginal bleeding, edema, severe headache and convulsions during pregnancy relative to nonsmokers. Mortality per 100,000 pregnancies appeared to be higher among smokers than nonsmokers in the placebo group (915 vs 584, RR=1.57, 95% CI: 0.80-3.08). beta-Carotene supplementation reduced pregnancy-related mortality both among smokers (RR=0.31 95% CI: 0.11 0.89) and nonsmokers (RR=0.41, 95% CI: 0.19-0.89). Similar results obtained with vitamin A supplementation were not statistically significant. Infant mortality up to 6 months was approximately 30% higher among smokers compared to nonsmokers in the placebo group both before and after adjusting for confounding factors. Neither supplement given to women reduced infant mortality. CONCLUSIONS: Cigarette smoking during pregnancy is associated with an increased risk of maternal and infant mortality in rural Nepal. beta-Carotene and to some extent vitamin A may reduce the risk of pregnancy-related mortality, but not infant mortality, among both smokers and nonsmokers. PMID- 14749739 TI - Relation between food intake and visual analogue scale ratings of appetite and other sensations in healthy older and young subjects. AB - OBJECTIVE: Visual analogue scales are widely used in appetite research, yet the validity of these scales to evaluate appetite and mood has not been assessed in older subjects. The aim of this study was to determine the relations between food intake and visual analogue scale (VAS) ratings of appetite and nonappetite sensations in healthy older and young subjects. DESIGN: Retrospective combined analysis of four single-blind, randomised, controlled appetite studies. SETTING: All studies were conducted in the University of Adelaide, Department of Medicine, Adelaide, Australia. SUBJECTS: A total of 45 healthy young men (n=24) and women (n=21) aged 18-35 y and 45 healthy older men (n=24) and women (n=21) aged 65-85 y were recruited by advertisement. INTERVENTIONS: Oral, intraduodenal or intravenous administration of treatments which suppressed food intake were compared to control. Up to 90 min after treatment, a test meal was offered and subjects ate freely for between 30 and 60 min. Perceptions were assessed by 100 mm visual analogue scales administered at regular intervals. RESULTS: Food intake at the test meal was positively related to perceptions of hunger, drowsiness, and calmness at both baseline and premeal (r>0.16, P<0.05), and inversely related to premeal ratings of fullness (r> 0.2, P<0.05) in both older and young subjects. Food intake was related to VAS ratings at least as strongly, if not more so, in older as in young subjects. CONCLUSIONS: These observations (i) confirm that food intake is related to perceptions of hunger and fullness as assessed by VAS in healthy older and young subjects, and (ii) suggest that sensations, not obviously associated with appetite, including 'drowsiness' and 'calmness', are also associated with food intake. PMID- 14749740 TI - Virtual food components: functional food effects expressed as food components. AB - BACKGROUND: The ability to communicate food benefits is essential for the successful development of functional foods and their role in improving public health. However, the functional efficacy of foods often cannot be represented by food composition. The concept of virtual food components (VFCs)-food data that express health-related effects, properties or functions of foods in the format of food components-is therefore proposed. OBJECTIVE: To develop protocols for designing VFC data sets that communicate functional efficacies of foods to end users, in order to facilitate evidence-based food choice, and allow data management systems to provide a more complete description of nutritional effects of foods than has been possible with values for actual food components alone. METHOD: A framework within which to develop VFCs was constructed, linking food choice to health end points. It involves scientific validation, generation of relative indices, their translation into a meaningful language based on equivalents to known and understood reference foods, followed by data consolidation and ecological validation. Criteria used to evaluate VFCs were importance, independence, validity, accuracy, robustness, sensitivity, linearity/additivity, relevance, comprehensiveness, acquirability, completeness, meaningfulness, acceptability and safety. The developmental framework and evaluative criteria were applied to glycaemic glucose equivalents (GGE), a VFC representing postprandial glycaemia, and to wheat bran equivalents for faecal bulk (WBE(fb)), a VFC representing faecal bulking efficacy. RESULTS: VFCs were used to identify foods according to health-related effects that cannot be accurately predicted from food composition data, and were used in a nutrition management system to concurrently show nutrient intake and physiological effects in the same units. The proposed evaluative criteria identified points requiring further research, and showed that lack of integrity-tested VFC data is an immediate challenge. CONCLUSION: VFCs are a means of communicating relative functional efficacy of foods as a continuous variable, and provide end users with a more accurate and complete view of the health effects of foods than can be provided by health claims or food composition data alone. PMID- 14749741 TI - Diet during pregnancy in relation to maternal weight gain and birth size. AB - OBJECTIVE: Maternal weight gain has been consistently linked to birth weight but, beyond maternal energy intake, no macronutrient has been associated with either of them. We have examined whether maternal energy-adjusted intake of macronutrients is associated with either maternal weight gain or birth-size parameters. DESIGN: Cohort study. SETTING: University hospital in Boston, USA. SUBJECTS: A total of 224 pregnant women coming for their first routine prenatal visit. The women were followed through delivery. INTERVENTIONS: None. Pregnant women's dietary intake during the second trimester was ascertained at the 27th week of pregnancy through a food frequency questionnaire. RESULTS: Intake of neither energy nor any of the energy-generating nutrients was significantly associated with birth size. In contrast, maternal weight gain by the end of the second trimester of pregnancy was significantly associated with energy intake (+0.9 kg/s.d. of intake; P approximately 0.006) as well as energy-adjusted intake of protein (+3.1 kg/s.d. of intake; P<10(-4)), lipids of animal origin (+2.6 kg/s.d. of intake; P<10(-4)) and carbohydrates (-5.2 kg/s.d. of intake; P<10( 4)). CONCLUSIONS: Although maternal weight gain is strongly associated with birth size, the indicated nutritional associations with weight gain are not reflected in similar associations with birth-size parameters. The pattern is reminiscent of the sequence linking diet to coronary heart disease (CHD) through cholesterol: diet has been conclusively linked to blood cholesterol levels and cholesterol levels are conclusively linked to this disease, even though the association of diet with CHD has been inconclusive and controversial. PMID- 14749742 TI - Decreased glutathione in patients with anorexia nervosa. Risk factor for toxic liver injury? AB - OBJECTIVE: To investigate glutathione and amino acids related to glutathione metabolism in patients with anorexia nervosa in order to test the hypothesis that these patients exhibit a deficiency of glutathione and therefore might be at an increased risk of developing toxic liver injury. DESIGN: Controlled observatory study and case report. SETTING: University Hospital. SUBJECTS: Subjects included 11 female patients with anorexia nervosa and 12 healthy female controls. INTERVENTIONS: Determination of fasting free and total glutathione, homocysteine, vitamins B(6) and B(12) and folic acid in plasma. RESULTS: A 14-y-old patient with a body mass index of 12.6 kg/m(2) presented with markedly elevated transaminases (ALAT >50 x upper limit of normal), and paracetamol was detected in her blood. Patients with anorexia nervosa exhibited lower circulating concentrations of free cysteine (8.9+/-1.5 vs 12.0+/-1.4 micromol/l) and free and total glutathione (5.0+/-1.3 vs 7.1+/-1.2 and 11.2+/-3.8 vs 16.2+/-5.0 micromol/l, respectively). The plasma concentrations of homocysteine (17.5+/-4.9 vs 12.0+/-3.8 micromol/l) and also of glycine (194+/-37 vs 143+/-41 micromol/l) and glutamine (422+/-51 vs 353+/-51 micromol/l) were significantly higher in patients with anorexia nervosa who were not deficient in folic acid, vitamin B(6) and B(12). CONCLUSIONS: Lower plasma concentrations of glutathione suggest lower intracellular concentrations of the tripeptide. Higher homocysteine, glycine and glutamine concentrations point to a decreased utilization of these amino acids for glutathione synthesis and an impairment of trans-sulfuration. Consequently, the capacity of patients with anorexia nervosa to detoxify electrophilic metabolites and reactive oxygen species via glutathione may be impaired. PMID- 14749743 TI - Effects of elderberry juice on fasting and postprandial serum lipids and low density lipoprotein oxidation in healthy volunteers: a randomized, double-blind, placebo-controlled study. AB - BACKGROUND: In a recent pilot study, the intake of elderberry juice resulted in a significant decrease in serum cholesterol concentrations and an increase in low density lipoprotein (LDL) stability. This study was designed to verify the preliminary results. OBJECTIVE: We investigated the impact of elderberry juice on cholesterol and triglyceride concentrations as well as antioxidant status in a cohort of young volunteers. DESIGN: Study A: The randomized, placebo-controlled trial for studying the effect of anthocyanes on lipid and antioxidant status, 34 subjects took capsules with 400 mg spray-dried powder containing 10% anthocyanes t.i.d. equivalent to 5 ml elderberry juice for 2 weeks. A subgroup of 14 subjects continued for an additional week to test for resistance to oxidation of LDL. Study B: To investigate the short-term effects on serum lipid concentrations, six subjects took a single dose of 50 ml of elderberry juice (equivalent to 10 capsules) along with a high-fat breakfast. RESULTS: In the placebo-controlled study, there was only a small, statistically not significant change in cholesterol concentrations in the elderberry group (from 199 to 190 mg/dl) compared to the placebo group (from 192 to 196 mg/dl). The resistance to copper induced oxidation of LDL did not change within 3 weeks. In the single-dose experiment increases in postprandial triglyceride concentrations were not significantly different when the six subjects were investigated with and without elderberry juice. CONCLUSIONS: Elderberry spray-dried extract at a low dose exerts a minor effect on serum lipids and antioxidative capacity. Higher, but nutritionally relevant doses might significantly reduce postprandial serum lipids. PMID- 14749744 TI - Replacing cows' with sheep's dairy fat lowers plasma cholesterol concentration in participants consuming dairy fat-rich diets. AB - OBJECTIVE: To determine the effects on plasma cholesterol concentration of replacing cows' dairy fat with sheep's dairy fat. DESIGN: Randomised crossover dietary intervention. SETTING: General community, Dunedin, New Zealand. SUBJECTS: Volunteer sample of 41 healthy adults with initial plasma cholesterol concentration between 4.8 and 7.8 mmol/l. INTERVENTIONS: Participants were asked to follow a self-selected low-fat background diet throughout the study to which, during each of the 2, 3-week dairy diets, they were asked to add sheep's or cows' dairy products. MAIN OUTCOME MEASURES: Energy and nutrient intakes, plasma triacylglycerol fatty acids, and plasma cholesterol. RESULTS: Energy and nutrient intakes on the sheep-dairy and cow-dairy diets were very similar, with total, saturated, monounsaturated and polyunsaturated fat contributing 34, 18-19, 9, and 3% of total energy intake, respectively. Participants consumed approximately 50 g/day of dairy fat on each diet. Replacing cows' with sheep's dairy fat led to a 0.33 (0.11-0.56, 95% CI) mmol/l decrease (6%) in plasma total cholesterol concentration, from 5.53 (0.90, s.d.) to 5.20 (0.90) mmol/l. Plasma low-density lipoprotein (LDL) cholesterol was 0.18 (0.02-0.33) mmol/l lower on the sheep dairy diet as was the concentration of plasma high-density lipoprotein (HDL) cholesterol, 0.11 (0.02-0.20) mmol/l. The LDL to HDL cholesterol ratio at the end of the sheep-dairy diet, 2.91 (1.10), was not significantly different (P>0.05) from the cow-dairy diet, 2.73 (0.83). CONCLUSIONS: Within the context of a diet high in dairy fat (50 g/day), replacing cows' milk fat with sheep's milk fat leads to a small reduction in plasma cholesterol concentration, but no change in the ratio of LDL to HDL cholesterol. PMID- 14749745 TI - What are the key food groups to target for preventing obesity and improving nutrition in schools? AB - OBJECTIVE: To determine differences in the contribution of foods and beverages to energy consumed in and out of school, and to compare consumption patterns between school canteen users and noncanteen users. DESIGN: Cross-sectional National Nutrition Survey, 1995. SETTING: Australia. SUBJECTS ON SCHOOL DAYS: A total of 1656 children aged 5-15 y who had weekday 24-h dietary recall data. RESULTS: An average of 37% of total energy intake was consumed at school. Energy-dense foods and beverages such as fat spreads, packaged snacks, biscuits and fruit/cordial drinks made a greater contribution to energy intake at school compared to out of school (P< or =0.01). Fast foods and soft drinks contributed 11 and 3% of total energy intake; however, these food groups were mostly consumed out of school. Fruit intake was low and consumption was greater in school. In all, 14% of children purchased food from the canteen and they obtained more energy from fast food, packaged snacks, desserts, milk and confectionary (P< or =0.05) than noncanteen users. CONCLUSIONS: : Energy-dense foods and beverages are over represented in the Australian school environment. To help prevent obesity and improve nutrition in schools, biscuits, snack bars and fruit/cordial drinks brought from home and fast food, packaged snacks, and confectionary sold at canteens should be replaced with fruit and water. PMID- 14749746 TI - Relation between calcium absorption and serum calcitriol in normal men: evidence for age-related intestinal resistance to calcitriol. AB - OBJECTIVE: To obtain information on the causes of age-related bone loss in men and the concomitant decline in calcium absorption. DESIGN: Cross-sectional study. SETTING: Adelaide, South Australia, Australia. SUBJECTS: A total of 95 healthy, Caucasian men (age range 27-87 y). RESULTS: Calcium absorption declined with age (r=-0.46, P<0.0001), as did 24-h urine calcium, phosphate and creatinine (r> 0.21, P<0.05 for all); serum calcitriol and 25 hydroxyvitamin D did not change with age. Calcium absorption was related to serum calcitriol (r=0.20, P=0.05). An inverse relation between the residual deviations in calcium absorption, after allowing for its dependence on calcitriol, and age (F=5.4, P<0.005) was observed. The 24-h urinary calcium, phosphate and creatinine were all related to calcium absorption (r>0.41, P<0.0001). Forearm bone density fell with age (r=-0.45, P<0.0001) but was not related to calcium absorption, or markers of bone turnover. CONCLUSIONS: In healthy Caucasian males (i) calcium absorption falls, but serum calcitriol does not change with age, (ii) the relation between calcium absorption and serum calcitriol changes with age, indicative of an intestinal resistance to calcitriol and (iii) calcium absorption is a significant determinant of 24-h urinary calcium excretion. PMID- 14749747 TI - Influence of a mineral water rich in calcium, magnesium and bicarbonate on urine composition and the risk of calcium oxalate crystallization. AB - OBJECTIVE: To evaluate the effect of a mineral water rich in magnesium (337 mg/l), calcium (232 mg/l) and bicarbonate (3388 mg/l) on urine composition and the risk of calcium oxalate crystallization. DESIGN: A total of 12 healthy male volunteers participated in the study. During the baseline phase, subjects collected two 24-h urine samples while on their usual diet. Throughout the control and test phases, lasting 5 days each, the subjects received a standardized diet calculated according to the recommendations. During the control phase, subjects consumed 1.4 l/day of a neutral fruit tea, which was replaced by an equal volume of a mineral water during the test phase. On the follow-up phase, subjects continued to drink 1.4 l/day of the mineral water on their usual diet and collected 24-h urine samples weekly. RESULTS: During the intake of mineral water, urinary pH, magnesium and citrate excretion increased significantly on both standardized and normal dietary conditions. The mineral water led to a significant increase in urinary calcium excretion only on the standardized diet, and to a significantly higher urinary volume and decreased supersaturation with calcium oxalate only on the usual diet. CONCLUSIONS: The magnesium and bicarbonate content of the mineral water resulted in favorable changes in urinary pH, magnesium and citrate excretion, inhibitors of calcium oxalate stone formation, counterbalancing increased calcium excretion. Since urinary oxalate excretion did not diminish, further studies are necessary to evaluate whether the ingestion of calcium-rich mineral water with, rather than between, meals may complex oxalate in the gut thus limiting intestinal absorption and urinary excretion of calcium and oxalate. PMID- 14749748 TI - Habitual fish consumption and glycated haemoglobin: the EPIC-Norfolk study. AB - OBJECTIVE: To investigate the association between habitual fish consumption and a continuous measure of glycaemia. DESIGN: Cross-sectional study. SETTING: EPIC Norfolk, a population-based cohort study of diet and chronic disease. SUBJECTS AND METHODS: In all, 4500 men and 5509 women, aged 40-78 y, without self-reported diabetes. Diet was assessed by a semiquantitative food frequency questionnaire, and glycaemia was measured by glycated haemoglobin. RESULTS: In women only, in analyses adjusted for age, the HbA(1c) level was positively associated with eating fried fish and inversely associated with eating oily fish (b=0.036, 95% confidence interval (CI): 0.0033, 0.069; and b=-0.046, 95% CI:-0.086, -0.0064 respectively). These associations were attenuated by adjustment for family history of diabetes, smoking status and physical activity level, but the association with fried fish remained statistically significant (b=0.033, 95% CI: 0.00056, 0.066). Adjusting for total energy, alcohol, fruit and vegetable intakes resulted in further attenuation and both associations were no longer statistically significant. In men, there was no evidence that HbA(1c) level was associated with fish consumption. CONCLUSIONS: The study found no evidence of an association between fish consumption and HbA(1c) after taking other lifestyle factors into account. PMID- 14749749 TI - Habitual physical activity and physical activity intensity: their relation to body composition in 5.0-10.5-y-old children. AB - BACKGROUND: Concerns of a decrease in physical activity levels (PALs) of children and a concurrent increase in childhood obesity exist worldwide. The exact relation between these two parameters however has as yet to be fully defined in children. OBJECTIVE: This study examined the relation in 47 children, aged 5-10.5 y (mean age 8.4+/-0.9 y) between habitual physical activity, minutes spent in moderate, vigorous and hard intensity activity and body composition parameters. DESIGN: Total energy expenditure (TEE) was calculated using the doubly labelled water technique and basal metabolic rate (BMR) was predicted from Schofield's equations. PAL was determined by PAL=TEE/BMR. Time spent in moderate, vigorous and hard intensity activity was determined by accelerometry, using the Tritrac R3D. Body fatness and body mass index (BMI) were used as the two measures of body composition. RESULTS: Body fat and BMI were significantly inversely correlated with PAL (r=-0.43, P=0.002 and r=-0.45, P=0.001). Times spent in vigorous activity and hard activity were significantly correlated to percentage body fat (r=-0.44, P=0.004 and r=-0.39, P=0.014), but not BMI. Children who were in the top tertiles for both vigorous activity and hard activity had significantly lower body fat percentages than those in the middle and lowest tertiles. Moderate intensity activity was not correlated with measures of body composition. CONCLUSIONS: As well as showing a significant relation between PAL and body composition, these data intimate that there may be a threshold of intensity of physical activity that is influential on body fatness. In light of world trends showing increasing childhood obesity, this study supports the need to further investigate the importance of physical activity for children. PMID- 14749750 TI - Nutritional status, growth and sleep habits among Senegalese adolescent girls. AB - OBJECTIVE: To study the relation between sleep habits, nutritional status, growth and maturation in a group of African adolescent girls. The main hypothesis to be tested was that sleep length could be an effective way to spare energy, and thus malnourished girls sleep longer than normal girls. DESIGN: Three repeated yearly surveys (1997-1999) on a subsample of girls drawn from a larger study cohort on growth at adolescence. SETTING: The Niakhar district in the central part of Senegal. SUBJECT: In total, 40 girls were initially drawn. Missing girls were replaced at each round by girls having the same characteristics and belonging to the same cohort. INTERVENTION: At each round, data on pubertal development (breast stages and occurrence of menarche), growth and nutritional status were collected. Adolescents wore an accelerometer for three or four consecutive nights and days at each round. RESULTS: At the beginning of the survey, girls were 13.3+/-0.5 y old. They were under international reference values in weight and height. Their mean sleep duration was 8.5+/-0.9 h. Their puberty status did not influence their sleep habits; however, they slept more in March than in June, which was related to the seasonal change in daylight. There was a significant relation between body mass index and sleep habits: thinner girls slept a longer time and more quietly than the more corpulent girls. CONCLUSION: The nutritional status of these girls influenced their sleep habits: this may have been either a direct causal relation or a consequence of a protective attitude on the part of the mothers towards the frailer girls. PMID- 14749751 TI - Population dietary habits and physical activity modification with age. AB - OBJECTIVE: The aim of the present study was to analyse the relation between age and both dietary habits and leisure-time physical activity, and to determine nutrient inadequacy of aged groups in our population. DESIGN: Cross-sectional study. SETTING: A random sample of the 25-74-y-old population of Gerona, Spain. SUBJECTS: A total of 838 men and 910 women were selected from among the general population according to the 1991 census. OUTCOME MEASURES: Analysis of dietary habits, including amount and type of alcohol consumption, and detailed evaluation of leisure-time physical activity. RESULTS: Nutrient densities of carbohydrates, vitamin B(1), vitamin B(12), vitamin C, vitamin E, folate, potassium, iron, magnesium, copper, and dietary fiber increased significantly (P<0.05) with age in both genders, whereas an inverse trend was observed for total fat, saturated fatty acids, cholesterol, and sodium. Multiple linear regression analysis revealed a direct association of healthy dietary habits, characterized through a composite dietary score, with age after adjusting for several confounders both in men and women (P<0.001). This score was composed of folate, vitamin C, vitamin E, beta-carotene, dietary fibre, cholesterol, saturated fatty acids, and sodium. In all, 29 and 10% of male and female subjects aged 65-74 y, respectively, reported inadequate intakes of six or more of 16 nutrients. Total leisure-time physical activity increased with age in men (P<0.002), and was not different among female age groups. CONCLUSION: Dietary behaviours and levels of physical activity spent during leisure time indicate a healthy lifestyle of the aged men and women in the present population. Nutrient inadequacy observed in some aged men and women, however, deserves particular intervention of health-care programmes for this growing part of our society. PMID- 14749752 TI - Insulin sensitivity in Chinese ovo-lactovegetarians compared with omnivores. AB - AIM: To compare the insulin sensitivity indices between Chinese vegetarians and omnivores. METHODS: The study included 36 healthy volunteers (vegetarian, n=19; omnivore, n=17) who had normal fasting plasma glucose levels. Each participant completed an insulin suppression test. We compared steady-state plasma glucose (SSPG), fasting insulin, the homeostasis model assessment for insulin sensitivity (HOMA-IR and HOMA %S) and beta-cell function (HOMA %beta) between the groups. We also tested the correlation of SSPG with years on a vegetarian diet. RESULTS: The omnivore subjects were younger than the vegetarians (55.7+/-3.7 vs 58.6+/-3.6 year of age, P=0.022). There was no difference between the two groups in sex, blood pressure, renal function tests and lipid profiles. The omnivores had higher serum uric acid levels than vegetarians (5.25+/-0.84 vs 4.54+/-0.75 mg/dl, P=0.011). The results of the indices were different between omnivores and vegetarians (SSPG (mean+/-s.d.) 105.4+/-10.2 vs 80.3+/-11.3 mg/dl, P<0.001; fasting insulin, 4.06+/-0.77 vs 3.02+/-1.19 microU/ml, P=0.004; HOMA-IR, 6.75+/ 1.31 vs 4.78+/-2.07, P=0.002; HOMA %S, 159.2+/-31.7 vs 264.3+/-171.7%, P=0.018) except insulin secretion index, HOMA %beta (65.6+/-18.0 vs 58.6+/-14.8%, P=0.208). We found a clear linear relation between years on a vegetarian diet and SSPG (r=-0.541, P=0.017). CONCLUSIONS: The vegetarians were more insulin sensitive than the omnivore counterparts. The degree of insulin sensitivity appeared to be correlated with years on a vegetarian diet. PMID- 14749753 TI - Lymphocyte proliferation and apoptosis in HIV-seropositive and healthy subjects during long-term ingestion of fruit juices or a fruit-vegetable-concentrate rich in polyphenols and antioxidant vitamins. AB - OBJECTIVE: We investigated whether ingestion of polyphenols from fruit juices or a fruit-vegetable-concentrate affects lymphocyte proliferation and apoptosis in human immunodeficiency virus (HIV)-seropositive (HIV(+)) and HIV-seronegative (HIV(-)) subjects. DESIGN: Randomized, prospective pilot intervention study. SETTING: University of Bonn, Department of General Internal Medicine. SUBJECTS: A total of 23 HIV(+) subjects from the HIV outpatient clinic, 18 HIV(-) controls. INTERVENTIONS: Subjects ingested either 1 l of fruit juice or 30 ml of fruit vegetable-concentrate daily for 16 weeks in addition to their regular diet. Lymphocyte proliferation and apoptosis were investigated in peripheral blood mononuclear cells at baseline, during 16-weeks of intervention, and after a 6 week washout. Proliferation was assessed by (3)H-thymidine incorporation and apoptosis by nuclear content as measured by flow cytometry. RESULTS: Supplementation of fruit juices increased phytohemagglutinin-induced lymphocyte proliferation (mitotic index) in HIV(+) patients from 18+/-16 to 40+/-34 (P=0.004) and in healthy controls from 27+/-16 to 51+/-21 (P=0.016). Apoptosis was not affected in HIV(+) patients, but rose in healthy controls from 9+/-10 to 34+/-11 (apoptotic index; P=0.001). Intervention with concentrate did not significantly alter proliferation and apoptosis in HIV(+) and HIV(-) subjects. CONCLUSIONS: Even though apoptosis did not change in HIV(+) subjects, ingestion of polyphenol-rich fruit juices might be favorable to HIV(+) patients due to enhanced proliferation, which could restore disturbances in T-cell homeostasis. In healthy controls, increased lymphocyte proliferation during juice consumption was counterbalanced by increased apoptosis. PMID- 14749754 TI - Micronutrient intake in overweight subjects is not deficient on an ad libitum fat reduced, high-simple carbohydrate diet. AB - OBJECTIVE: To investigate whether subjects consuming a fat-reduced, high-simple carbohydrate diet (SCHO) are at greater risk of micronutrient inadequacy than subjects consuming a fat-reduced, high-complex carbohydrate (CCHO) or a normal fat diet (control, CD). DESIGN: A 6-month randomised controlled dietary intervention trial with a parallel design. METHODS: In total, 46 overweight (BMI: 24.4-36.3 kg/m(2)) subjects (19 males, 27 females) aged 21-54 y consumed one of three ad libitum diets: SCHO, CCHO, or CD. Nutrient intake was assessed by a 7 day weighed food record. RESULTS: Self-reported energy intake did not differ between diet groups. The lowest intake of vitamin B(12) was found in the SCHO group vs CCHO (P=0.025) and vs. CD (P=0.012). In men, zinc intake was lower on the SCHO diet compared to the CD diet (P=0.018). The recommendations for zinc and vitamin B(12) were, however, met by all the diet groups. No other diet differences were observed. Intake of several micronutrients were insufficient in all three diet groups, although in most cases comparable to average Danish intakes. CONCLUSION: Zinc intake in men and vitamin B(12) intake in the combined gender groups were lower on a fat-reduced, simple carbohydrate-rich diet compared to a habitual, normal-fat diet, but not below recommended levels. PMID- 14749755 TI - High prevalence of vitamin D insufficiency in healthy elderly people living at home in Argentina. AB - OBJECTIVE: To evaluate the nutritional status of vitamin D in urban populations of healthy elderly people living at home, in different regions of Argentina. DESIGN: Cross-sectional study. SUBJECTS: In total, 386 ambulatory subjects over 65 y of age from seven cities (between latitude 26 degrees S and 55 degrees S) were asked to participate between the end of winter and the beginning of spring. Of these, 369 accepted, 30 were excluded because of medical history or abnormal biochemical determinations. Finally, 339 subjects (226 women and 113 men) (X+/ s.d.) (71.3+/- 5.2 y) were included. RESULTS: Serum 25OHD levels were lowest in the South (latitude range: 41 degrees S-55 degrees S): 14.2+/-5.6 ng/ml (P<0.0001vs North and Mid regions); highest in the North (26 degrees S-27 degrees S): 20.7+/-7.4 ng/ml (P<0.03 vs Mid, P<0.0001vs South); and intermediate in the Mid region (33 degrees S-34 degrees S) 17.9+/-8.2 ng/ml. Serum mid-molecule PTH (mmPTH) and 25OHD were inversely related: (r=-0.24, P<0.001). A cutoff level of 25OHD at which serum mmPTH levels began to increase was established at 27 ng/ml. A high prevalence (87-52%) of subjects with 25OHD levels in the deficiency insufficiency range (25OHD levels <20 ng/ml) was detected. CONCLUSION: This study shows that vitamin D deficiency/insufficiency in the elderly is a worldwide problem. Correction of this deficit would have a positive impact on bone health of elderly people. PMID- 14749756 TI - Bioavailability of selenium from fish, yeast and selenate: a comparative study in humans using stable isotopes. AB - OBJECTIVE: To measure the bioavailability of selenium from cooked and raw fish in humans by estimating and comparing apparent absorption and retention of selenium in biosynthetically labelled fish with labelled selenate and biosynthetically labelled selenium in brewers yeast. DESIGN: The intervention study was a parallel, randomised, reference substance controlled design carried out at two different centres in Europe. SETTING: The human study was carried out at the Institute of Food Research, Norwich, UK and at TNO Nutrition and Food Research, Zeist, The Netherlands. SUBJECTS: In all, 35 male volunteers aged 18-50 y were recruited; 17 subjects were studied in Norwich (UK) and 18 in Zeist (Netherlands). All of the recruited subjects completed the study. INTERVENTIONS: Biosynthetically labelled trout fish (processed by two different methods), biosynthetically labelled brewers yeast and isotopically labelled selenate were used to estimate selenium apparent absorption and retention by quantitative analysis of stable isotope labels recovered in faeces and urine. Subjects consumed the labelled foods in four meals over two consecutive days and absorption was measured by the luminal disappearance method over 10 days. Urinary clearance of isotopic labels was measured over 7 days to enable retention to be calculated. RESULTS: Apparent absorption of selenium from fish was similar to selenate and there was no difference between the two processing methods used. However, retention of fish selenium was significantly higher than selenate (P<0.001). Apparent absorption and retention of yeast selenium was significantly different (P<0.001) from both fish selenium and selenate. CONCLUSION: Fish selenium is a highly bioavailable source of dietary selenium. Cooking did not affect selenium apparent absorption or retention from fish. Selenium from yeast is less bioavailable. PMID- 14749757 TI - Bioavailability of selenium from bovine milk as assessed in subjects with ileostomy. AB - OBJECTIVE: To assess the absorption of dietary selenium in humans, especially of milk selenium. DESIGN: : 1-day meal studies in subjects with ileostomy. SETTING: Hospital outpatient clinics. SUBJECTS: Three subjects in the pilot study and nine subjects in the main study (eight men/ four women). INTERVENTION: Different beverages, 1 l/day, were given in addition to basal diets (soft drink, 1 week; low-fat milk, 3 weeks; fermented low-fat milk, 3 weeks and soft drink, 1 week). Ileostomy effluents were collected during the last 2 days in each of the four periods. RESULTS: On days when the subjects were given 1 l of low-fat milk, the estimated fractional absorption of total dietary selenium was 65.5 (2.3)% (mean (s.d.), n=18), which was similar to the value when fermented low-fat milk was given (64.1 (3.2)%). However, both the calculated amount of milk selenium absorbed (10.9 (2.4) vs 9.4 (1.7) microg selenium) and its fractional absorption (73.3 (16.1) vs 64.1 (11.2)%, n=18) were significantly higher for milk than for fermented milk. CONCLUSIONS: Selenium from milk and other sources is well absorbed in subjects with ileostomy. The real absorption may be even higher than the values shown. PMID- 14749758 TI - Growth and feeding practices of Vietnamese infants in Australia. AB - OBJECTIVE: To investigate the growth and feeding practices in first-generation Vietnamese infants living in Australia. DESIGN: Cohort study. SETTING: The study was conducted between 1999 and 2002 in Sydney. SUBJECTS: A total of 239 Vietnamese women were recruited randomly from antenatal clinics, and of these 210 were initially seen. During the first year, 20 cases (9.5%) were lost to follow up. Data were collected at 0.5, 2, 4, 6, 9 and 12 months. RESULTS: Vietnamese infants were significantly longer and heavier than reference data (both P<0.0001). The Vietnamese infants had a significant decline in weight growth with age compared with reference data (P<0.001). The Vietnamese infants had marginally higher s.d. score for ideal weight for length than reference data (P=0.044). There was a significant decline in ideal weight for length with age compared with reference data (P=0.0065). Both parents were significantly shorter (mean s.d. height scores: -1.5+/-0.8 (mother) and -1.8+/-0.8 (father)) than reference data (P<0.001). The incidence of breast feeding was 79%, but half of the breast feeding women had stopped breast feeding by 3 months. A total of 162 (79.8%) infants were given infant formula within the first week, of whom 131 (80.1%) were fed infant formula within the first 24 h after birth. CONCLUSIONS: Vietnamese infants in this study had growth comparable with reference data despite their parents being shorter than reference data. Breast feeding duration was short with infant formula being introduced early. PMID- 14749759 TI - Zinc and vitamin A intake and status in a national sample of British young people aged 4-18 y. AB - OBJECTIVE: To examine zinc and vitamin A intake and status and associated dietary, socio-demographic, lifestyle and physiological factors in British young people. DESIGN: National Diet and Nutrition Survey of young people aged 4-18 y. SETTING: Great Britain, 1997. SUBJECTS: Complete 7-day weighed dietary records were provided by 1520 participants, while 1193 provided blood samples. RESULTS: A total of 13 and 11% of participants respectively reported low dietary intakes of zinc and vitamin A (retinol equivalents), relative to the UK lower reference nutrient intake. These percentages were not altered significantly by including contributions to intake from supplements, mainly containing vitamin A (as retinol). Likelihood of low zinc and/or vitamin A intake was more often associated with age, sex and likely under-reporting of food consumption than with other socio-demographic and lifestyle factors. Low zinc and vitamin A intakes were generally less likely in those with higher consumption of dairy foods (mainly milk). Zinc and vitamin A status (assessed by plasma zinc and retinol concentrations) were adequate in almost all participants. Plasma zinc concentration was not significantly associated with zinc intake. Plasma retinol concentration was correlated with vitamin A intake (overall r=0.17, P<0.001; adjusted for age and plasma alpha(1)-antichymotrypsin concentration) and increased significantly with age (P<0.001) in both sexes. A significant association was found between plasma zinc and retinol concentrations in boys only (r=0.17, P=0.001). CONCLUSION: Zinc and vitamin A intakes and status were generally adequate in this national sample of British young people. PMID- 14749760 TI - Mineral water fortified with folic acid, vitamins B6, B12, D and calcium improves folate status and decreases plasma homocysteine concentration in men and women. AB - OBJECTIVE: We investigated the effects of mineral water fortified with folic acid, vitamins B(6), B(12), D and calcium on folate concentrations in serum and erythrocytes, serum vitamin B(12) and plasma homocysteine concentrations in free living subjects. In addition, we investigated the bioavailability of calcium added to mineral water by measuring urinary calcium excretion and serum alkaline phosphatase activity. DESIGN: Randomized, controlled, double-blinded, parallel design. SETTING: Outpatient dietary intervention with free-living subjects in Eastern Finland. SUBJECTS: Altogether, 66 subjects were recruited for the study. In all, 60 subjects completed the study. INTERVENTIONS: The study began with a 2 week run-in period followed by an 8-week intervention period. During the intervention study, subjects consumed mineral water fortified with folic acid (563 microg/day), vitamins B(6) (1 mg/day), B(12) (7.5 microg/day), cholecalciferol (0.6 microg/day) and calcium (563 mg/day) or placebo mineral water. RESULTS: The fortified mineral water increased serum and erythrocyte folate concentrations by 16.1+/-5.6 nmol/l (P<0.001) and 199+/-76 nmol/l (P<0.001), respectively, and decreased plasma homocysteine concentration by 1.6 micromol/l (P<0.001). Urinary calcium excretion and serum alkaline phosphatase activity for 24 h increased significantly (P<0.001 and P=0.01 respectively) in the intervention group. CONCLUSIONS: Mineral water fortified with folic acid, vitamins B(6), B(12) and D and calcium enhanced folate status and reduced plasma homocysteine concentration in normohomocysteinemic subjects without folate deficiency. Indirect measures of calcium and bone metabolism indicated that the calcium used in the fortification of the mineral water was bioavailable. PMID- 14749761 TI - Drug use during breastfeeding. A survey from the Netherlands. AB - OBJECTIVE: To survey drug use by breastfeeding women, and to compare this with nonbreastfeeding women. In addition, we were interested whether drug use was of influence on the decision to give breastfeeding, and the other way around. DESIGN AND SETTING: During a 6-week period in 2002, a questionnaire was handed out to all women with a child not older than 6 months, who visited a Well-Baby Clinic in the province of Friesland, the Netherlands, eventually resulting in 549 returned questionnaires (response 43%). RESULTS: In all, 82.1% of the participants breastfed their baby at least at any time during the first 6 months after birth. More than half (65.9%) of all breastfeeding women had used drugs; however, they used drugs less frequently than nonbreastfeeding women (79.6%). The pattern of drug use differed: oral contraceptives, iron preparations, drugs for peptic ulcer, and several psychotropic drugs were more frequently used by nonbreastfeeding women, while vitamins were more frequently used by breastfeeding women. Drugs play an important role in women's decision to start or continue breastfeeding: women frequently hesitated to use drugs during breastfeeding, stopped either breastfeeding or drug use to avoid combining the two, took a measure to minimise exposure to the child, did not use any drug because of breastfeeding, or did not breastfeed because of drug use. CONCLUSIONS: Drugs are frequently though reluctantly used during breastfeeding, and play an important role in the decision to start and stop breastfeeding. Information how to deal with drugs seems therefore indispensable in efforts to promote breastfeeding. PMID- 14749763 TI - The microvesicle as a vehicle for EMMPRIN in tumor-stromal interactions. AB - EMMPRIN is a transmembrane glycoprotein expressed at high levels by tumor cells. It has been identified as a tumor-derived factor that can stimulate matrix metalloproteinase expression in fibroblasts and hence facilitate tumor invasion and metastasis. Recent studies have shown that full-length EMMPRIN is released by tumor cells, but the mechanism of release remains unclear. Here, we show that EMMPRIN is released from the surface of NCI-H460 cells via microvesicle shedding. However, these vesicles are unstable and rapidly break down to release bioactive EMMPRIN. Although microvesicle shedding has been considered a constitutive process in tumor cells, our data show that it can be amplified upon cell exposure to PMA, elucidating at least one signalling cascade responsible for EMMPRIN release. This pathway is dependent on protein kinase C, calcium mobilization and mitogen-activated protein kinase kinase (MEK 1/2). Thus, the results outline a novel form of tumor-stromal interaction in which extracellular matrix degradation by fibroblasts is controlled through the microvesicular release of EMMPRIN from tumor cells. PMID- 14749762 TI - Common alterations in gene expression and increased proliferation in recurrent acute myeloid leukemia. AB - Recurrent disease following high-dose chemotherapy is a major problem in patients with acute myeloid leukemia (AML). To identify its characteristics, we performed expression profiling in blasts from untreated AML and relapse, using a specific cDNA microarray comprising 4128 genes generated by cDNA subtraction supplemented with cancer-associated genes. Expression analysis of 18 AML bone marrow specimens showed that recurrent AML is commonly associated with the mRNA expression changes in a set of 58 genes. Increased cellular proliferation was indicated by the overexpression of the transferrin receptor, proliferating cell nuclear antigen, and G1 cyclins. An immunohistochemical study for Ki-67-positive blasts in 18 paired bone marrow biopsy samples confirmed a highly significant (P<0.0001) increase in the proliferation fraction at relapse. In addition, we found enhanced activation of the RAF/MEK/ERK cascade as mRNAs of MKP-1, c-jun, c-fos, and egr-1 were significantly increased at relapse. Immunohistochemistry and immunoblotting analyses for biphosphorylated ERK1/2 protein provide additional evidence for enhanced activation of the RAF/MEK/ERK pathway. The degree of increase is significantly correlated with the increased proliferation. Furthermore, the genes identified provide a rationale for further studies on predictive diagnosis and therapeutic intervention. PMID- 14749764 TI - Activation of the p53-dependent G1 checkpoint response in mouse embryo fibroblasts depends on the specific DNA damage inducer. AB - The p53 tumor suppressor protein inhibits proliferation by inducing either cell cycle arrest or apoptosis in response to cellular stresses. Mouse embryo fibroblasts (MEFs) provide a primary cell model system in which to examine both functions of p53. MEFs treated with gamma-rays undergo p53-dependent G1 arrest, while oncogene-expressing MEFs treated with a variety of DNA-damaging agents undergo p53-dependent apoptosis. Although the p53-dependent G1 arrest checkpoint response to gamma-rays in MEFs has been well characterized, the response to other DNA-damaging agents has not. Here, we examine the effects of commonly utilized chemotherapeutics, including doxorubicin, etoposide, and cisplatin, on cell cycle arrest in MEFs, and we define the p53 dependence of these effects. In addition, we examine the response of MEFs to ultraviolet light (UVC), as a representative agent acting by inducing pyrimidine dimers. Although p53 is clearly activated by all the agents examined, as measured by p21 induction, there are surprising differences in the activities of these agents. For example, doxorubicin but not cisplatin can effectively induce a p53-dependent G1 arrest. UVC, in contrast, induces a p53-independent G1 arrest response. Thus, the exact response of cells to DNA damage depends on the specific agent used. PMID- 14749765 TI - Genomic annotation of the meningioma tumor suppressor locus on chromosome 1p34. AB - Meningioma is a frequently occurring tumor of the meninges surrounding the central nervous system. Loss of the short arm of chromosome 1 (1p) is the second most frequent chromosomal abnormality observed in these tumors. Previously, we identified a 3.7 megabase (Mb) region of consistent deletion on 1p33-p34 in a panel of 157 tumors. Loss of this region was associated with advanced disease and predictive for tumor relapse. In this report, a high-resolution integrated map of the region was constructed (CompView) to identify all markers in the smallest region of overlapping deletion (SRO). A regional somatic cell hybrid panel was used to more precisely localize those markers identified in CompView as within or overlapping the region. Additional deletion mapping using microsatellites localized to the region narrowed the SRO to approximately 2.8 Mb. The 88 markers remaining in the SRO were used to screen genomic databases to identify large insert clones. Clones were assembled into a physical map of the region by PCR based, sequence-tagged site (STS) content mapping. A sequence from clones was used to validate STS content by electronic PCR and to identify transcripts. A minimal tiling path of 43 clones was constructed across the SRO. Sequence data from the most current sequence assembly were used for further validation. A total of 59 genes were ordered within the SRO. In all, 17 of these were selected as likely candidates based on annotation using Gene Ontology Consortium terms, including the MUTYH, PRDX1, FOXD2, FOXE3, PTCH2, and RAD54L genes. This annotation of a putative tumor suppressor locus provides a resource for further analysis of meningioma candidate genes. PMID- 14749766 TI - In praise of biochemistry. PMID- 14749767 TI - Insights into checkpoint capacity. PMID- 14749768 TI - Happy motoring with ATP synthase. PMID- 14749769 TI - Sizing up small RNAs. PMID- 14749770 TI - Barreling through the membrane. PMID- 14749771 TI - GTPase twins in the SRP family. PMID- 14749772 TI - A new deal for Holliday junctions. PMID- 14749774 TI - The enzymes and control of eukaryotic mRNA turnover. AB - The degradation of eukaryotic mRNAs plays important roles in the modulation of gene expression, quality control of mRNA biogenesis and antiviral defenses. In the past five years, many of the enzymes involved in this process have been identified and mechanisms that modulate their activities have begun to be identified. In this review, we describe the enzymes of mRNA degradation and their properties. We highlight that there are a variety of enzymes with different specificities, suggesting that individual nucleases act on distinct subpopulations of transcripts within the cell. In several cases, translation factors that bind mRNA inhibit these nucleases. In addition, recent work has begun to identify distinct mRNP complexes that recruit the nucleases to transcripts through different mRNA-interacting proteins. These properties and complexes suggest multiple mechanisms by which mRNA degradation could be regulated. PMID- 14749775 TI - Minimaxing risk. PMID- 14749776 TI - Tracking synapse-associated TCRs. PMID- 14749778 TI - Control freaks: immune regulatory cells. PMID- 14749779 TI - LIGHTing the way for tumor immunity. PMID- 14749780 TI - Right place, right time. PMID- 14749781 TI - Presenting fats with SAPs. PMID- 14749782 TI - Creating new peptide antigens by slicing and splicing proteins. PMID- 14749785 TI - Good and bad in Pakistan. PMID- 14749784 TI - Age-related changes in lymphocyte development and function. AB - The effects of aging on the immune system are widespread and extend from hematopoietic stem cells and lymphoid progenitors in the bone marrow and thymus to mature lymphocytes in secondary lymphoid organs. These changes combine to result in a diminution of immune responsiveness in the elderly. This review aims to provide an overview of age-related changes in lymphocyte development and function and discusses current controversies in the field of aging research. PMID- 14749787 TI - Prospects brighten around the red planet. PMID- 14749786 TI - Prolific ecologist vows to fight Danish misconduct verdict. PMID- 14749788 TI - Photo express. PMID- 14749790 TI - North Korea offers US tour party glimpse of weapons programme. PMID- 14749789 TI - Japan's ethnic crime database sparks fears over human rights. PMID- 14749791 TI - African labs win major role in tsetse-fly genome project. PMID- 14749792 TI - Quashed convictions reignite row over British cot deaths. PMID- 14749793 TI - Wildlife attacks hinder conservation efforts. PMID- 14749794 TI - NIH acts to quench 'conflict of interest' allegations. PMID- 14749796 TI - Mathematics: the reluctant celebrity. PMID- 14749797 TI - Brain development: the most important sexual organ. PMID- 14749799 TI - Hibben was not proved guilty of misconduct. PMID- 14749800 TI - Fraud offers big rewards for relatively little risk. PMID- 14749801 TI - Bridging a know--do gap. PMID- 14749802 TI - Biodefence funds have tight strings attached. PMID- 14749808 TI - Engineering complex systems. PMID- 14749809 TI - Fungi and the food of the gods. PMID- 14749810 TI - Nanotechnology: how does a nanofibre grow? PMID- 14749811 TI - Developmental biology: tail of decay. PMID- 14749813 TI - Earthquake science: faults greased at high speed. PMID- 14749814 TI - Materials science: a natural solution to corrosion? PMID- 14749815 TI - Cell biology: pathogen propulsion. PMID- 14749816 TI - Molecular motors: turning the ATP motor. PMID- 14749818 TI - Olfaction: scent-triggered navigation in honeybees. AB - The honeybee, Apis mellifera, navigates rapidly and accurately to food sources that are often kilometres away. They achieve this by learning visual cues, such as the location and colour of nectar-bearing flowers, and chemical cues, such as the scent and the taste of the nectar. Here we train bees to visit differently scented sugar feeders placed at specific outdoor locations and find that they can be induced to visit the same locations simply by having the corresponding scent blown into the hive, even when the destinations no longer have the food or carry the scent. A familiar nectar scent can trigger specific memories of a route and therefore expedite navigation to the food source. PMID- 14749819 TI - Lifespan: catch-up growth and obesity in male mice. AB - Poor fetal growth is linked with long-term detrimental effects on health in adulthood. Here we investigate whether the lifespan of male mice is affected by their growth rate when they were suckling and find that limiting growth during that period not only increases longevity but also protects against the life shortening effect of an obesity-inducing diet later on. By contrast, we find that lifespan is considerably shortened if the postnatal period of growth is accelerated to make up for reduced growth in utero, and that, in addition, these mice are susceptible to the adverse effects on longevity of an obesity-inducing diet after weaning. PMID- 14749820 TI - Palaeogeography: Devonian tetrapod from western Europe. AB - Several discoveries of Late Devonian tetrapods (limbed vertebrates) have been made during the past two decades, but each has been confined to one locality. Here we describe a tetrapod jaw of about 365 million years (Myr) old from the Famennian of Belgium, which is the first from western continental Europe. The jaw closely resembles that of Ichthyostega, a Famennian tetrapod hitherto known only from Greenland. The environment of this fossil provides information about the conditions that prevailed just before the virtual disappearance of tetrapods from the fossil record for 20 Myr. PMID- 14749821 TI - TNF ligands: is TALL-1 a trimer or a virus-like cluster? AB - Native TALL-1 (B-cell activation factor, BAFF; also known as BlyS) was initially described as a homotrimer, but Liu and colleagues claim that it is a 60-subunit complex on the basis of their results from X-ray crystallography and size exclusion chromatography. They consider TALL-1 60-mers to be the biologically active form, and the arrangement of the 60-mers resembles that of the capsid of satellite tobacco necrosis virus. Here we show that active TALL-1 is trimeric under normal physiological conditions and that formation of higher-order oligomers is an artefact of tagging the amino terminus of the protein with a histidine tag. PMID- 14749823 TI - Summing up the noise in gene networks. AB - Random fluctuations in genetic networks are inevitable as chemical reactions are probabilistic and many genes, RNAs and proteins are present in low numbers per cell. Such 'noise' affects all life processes and has recently been measured using green fluorescent protein (GFP). Two studies show that negative feedback suppresses noise, and three others identify the sources of noise in gene expression. Here I critically analyse these studies and present a simple equation that unifies and extends both the mathematical and biological perspectives. PMID- 14749824 TI - fgf8 mRNA decay establishes a gradient that couples axial elongation to patterning in the vertebrate embryo. AB - Formation and patterning of the vertebrate embryo occur in a head-to-tail sequence. This progressive mode of body formation from the posterior end of the embryo requires a strict temporal coordination of tissue differentiation--a process involving fibroblast growth factor (FGF) signalling. Here we show that transcription of fgf8 messenger RNA is restricted to the growing posterior tip of the embryo. fgf8 mRNA is progressively degraded in the newly formed tissues, resulting in the formation of an mRNA gradient in the posterior part of the embryo. This fgf8 mRNA gradient is translated into a gradient of FGF8 protein, which correlates with graded phosphorylation of the kinase Akt, a downstream effector of FGF signalling. Such a mechanism provides an efficient means to monitor the timing of FGF signalling, coupling the differentiation of embryonic tissues to the posterior elongation of the embryo. In addition, this mechanism provides a novel model for morphogen gradient formation. PMID- 14749825 TI - A high-mobility electron gas at the LaAlO3/SrTiO3 heterointerface. AB - Polarity discontinuities at the interfaces between different crystalline materials (heterointerfaces) can lead to nontrivial local atomic and electronic structure, owing to the presence of dangling bonds and incomplete atomic coordinations. These discontinuities often arise in naturally layered oxide structures, such as the superconducting copper oxides and ferroelectric titanates, as well as in artificial thin film oxide heterostructures such as manganite tunnel junctions. If polarity discontinuities can be atomically controlled, unusual charge states that are inaccessible in bulk materials could be realized. Here we have examined a model interface between two insulating perovskite oxides--LaAlO3 and SrTiO3--in which we control the termination layer at the interface on an atomic scale. In the simple ionic limit, this interface presents an extra half electron or hole per two-dimensional unit cell, depending on the structure of the interface. The hole-doped interface is found to be insulating, whereas the electron-doped interface is conducting, with extremely high carrier mobility exceeding 10,000 cm2 V(-1) s(-1). At low temperature, dramatic magnetoresistance oscillations periodic with the inverse magnetic field are observed, indicating quantum transport. These results present a broad opportunity to tailor low-dimensional charge states by atomically engineered oxide heteroepitaxy. PMID- 14749826 TI - Atomic-scale imaging of carbon nanofibre growth. AB - The synthesis of carbon nanotubes with predefined structure and functionality plays a central role in the field of nanotechnology, whereas the inhibition of carbon growth is needed to prevent a breakdown of industrial catalysts for hydrogen and synthesis gas production. The growth of carbon nanotubes and nanofibres has therefore been widely studied. Recent advances in in situ techniques now open up the possibility of studying gas-solid interactions at the atomic level. Here we present time-resolved, high-resolution in situ transmission electron microscope observations of the formation of carbon nanofibres from methane decomposition over supported nickel nanocrystals. Carbon nanofibres are observed to develop through a reaction-induced reshaping of the nickel nanocrystals. Specifically, the nucleation and growth of graphene layers are found to be assisted by a dynamic formation and restructuring of mono-atomic step edges at the nickel surface. Density-functional theory calculations indicate that the observations are consistent with a growth mechanism involving surface diffusion of carbon and nickel atoms. The finding that metallic step edges act as spatiotemporal dynamic growth sites may be important for understanding other types of catalytic reactions and nanomaterial syntheses. PMID- 14749827 TI - Enhanced ice sheet growth in Eurasia owing to adjacent ice-dammed lakes. AB - Large proglacial lakes cool regional summer climate because of their large heat capacity, and have been shown to modify precipitation through mesoscale atmospheric feedbacks, as in the case of Lake Agassiz. Several large ice-dammed lakes, with a combined area twice that of the Caspian Sea, were formed in northern Eurasia about 90,000 years ago, during the last glacial period when an ice sheet centred over the Barents and Kara seas blocked the large northbound Russian rivers. Here we present high-resolution simulations with an atmospheric general circulation model that explicitly simulates the surface mass balance of the ice sheet. We show that the main influence of the Eurasian proglacial lakes was a significant reduction of ice sheet melting at the southern margin of the Barents-Kara ice sheet through strong regional summer cooling over large parts of Russia. In our simulations, the summer melt reduction clearly outweighs lake induced decreases in moisture and hence snowfall, such as has been reported earlier for Lake Agassiz. We conclude that the summer cooling mechanism from proglacial lakes accelerated ice sheet growth and delayed ice sheet decay in Eurasia and probably also in North America. PMID- 14749828 TI - Natural examples of olivine lattice preferred orientation patterns with a flow normal a-axis maximum. AB - Tectonic plate motion is thought to cause solid-state plastic flow within the underlying upper mantle and accordingly lead to the development of a lattice preferred orientation of the constituent olivine crystals. The mechanical anisotropy that results from such preferred orientation typically produces a direction of maximum seismic wave velocity parallel to the plate motion direction. This has been explained by the existence of an olivine preferred orientation with an 'a-axis' maximum parallel to the induced mantle flow direction. In subduction zones, however, the olivine a axes have been inferred to be arranged roughly perpendicular to plate motion, which has usually been ascribed to localized complex mantle flow patterns. Recent experimental work suggests an alternative explanation: under conditions of high water activity, a 'B-type' olivine preferred orientation may form, with the a-axis maximum perpendicular to the flow direction. Natural examples of such B-type preferred orientation are, however, almost entirely unknown. Here we document widespread B type olivine preferred orientation patterns from a subduction-type metamorphic belt in southwest Japan and show that these patterns developed in the presence of water. Our discovery implies that mantle flow above subduction zones may be much simpler than has generally been thought. PMID- 14749829 TI - Friction falls towards zero in quartz rock as slip velocity approaches seismic rates. AB - An important unsolved problem in earthquake mechanics is to determine the resistance to slip on faults in the Earth's crust during earthquakes. Knowledge of coseismic slip resistance is critical for understanding the magnitude of shear stress reduction and hence the near-fault acceleration that can occur during earthquakes, which affects the amount of damage that earthquakes are capable of causing. In particular, a long-unresolved problem is the apparently low strength of major faults, which may be caused by low coseismic frictional resistance. The frictional properties of rocks at slip velocities up to 3 mm s(-1) and for slip displacements characteristic of large earthquakes have been recently simulated under laboratory conditions. Here we report data on quartz rocks that indicate an extraordinary progressive decrease in frictional resistance with increasing slip velocity above 1 mm s(-1). This reduction extrapolates to zero friction at seismic slip rates of approximately 1 m s(-1), and appears to be due to the formation of a thin layer of silica gel on the fault surface: it may explain the low strength of major faults during earthquakes. PMID- 14749830 TI - A new orang-utan relative from the Late Miocene of Thailand. AB - The fossil record of the living great apes is poor. New fossils from undocumented areas, particularly the equatorial forested habitats of extant hominoids, are therefore crucial for understanding their origins and evolution. Two main competing hypotheses have been proposed for orang-utan origins: dental similarities support an origin from Lufengpithecus, a South Chinese and Thai Middle Miocene hominoid; facial and palatal similarities support an origin from Sivapithecus, a Miocene hominoid from the Siwaliks of Indo-Pakistan. However, materials other than teeth and faces do not support these hypotheses. Here we describe the lower jaw of a new hominoid from the Late Miocene of Thailand, Khoratpithecus piriyai gen. et sp. nov., which shares unique derived characters with orang-utans and supports a hypothesis of closer relationships with orang utans than other known Miocene hominoids. It can therefore be considered as the closest known relative of orang-utans. Ancestors of this great ape were therefore evolving in Thailand under tropical conditions similar to those of today, in contrast with Southern China and Pakistan, where temperate or more seasonal climates appeared during the Late Miocene. PMID- 14749831 TI - The strict anaerobe Bacteroides fragilis grows in and benefits from nanomolar concentrations of oxygen. AB - Strict anaerobes cannot grow in the presence of greater than 5 micro M dissolved oxygen. Despite this growth inhibition, many strict anaerobes of the Bacteroides class of eubacteria can survive in oxygenated environments until the partial pressure of O2 (PO2) is sufficiently reduced. For example, the periodontal pathogens Porphyromonas gingivalis and Tannerella forsythensis colonize subgingival plaques of mammals, whereas several other Bacteroides species colonize the gastrointestinal tract of animals. It has been suggested that pre colonization of these sites by facultative anaerobes is essential for reduction of the PO2 and subsequent colonization by strict anaerobes. However, this model is inconsistent with the observation that Bacteroides fragilis can colonize the colon in the absence of facultative anaerobes. Thus, this strict anaerobe may have a role in reduction of the environmental PO2. Although some strictly anaerobic bacteria can consume oxygen through an integral membrane electron transport system, the physiological role of this system has not been established in these organisms. Here we demonstrate that B. fragilis encodes a cytochrome bd oxidase that is essential for O2 consumption and is required, under some conditions, for the stimulation of growth in the presence of nanomolar concentrations of O2. Furthermore, our data suggest that this property is conserved in many other organisms that have been described as strict anaerobes. PMID- 14749832 TI - A newly discovered Roseobacter cluster in temperate and polar oceans. AB - Bacterioplankton phylotypes of alpha-Proteobacteria have been detected in various marine regions, but systematic biogeographical studies of their global distribution are missing. Alpha-Proteobacteria comprise one of the largest fractions of heterotrophic marine bacteria and include two clades, SAR11 and Roseobacter, which account for 26 and 16% of 16S ribosomal RNA gene clones retrieved from marine bacterioplankton. The SAR11 clade attracted much interest because related 16S rRNA gene clones were among the first groups of marine bacteria to be identified by cultivation-independent approaches and appear to dominate subtropical surface bacterioplankton communities. Here we report on the global distribution of a newly discovered cluster affiliated to the Roseobacter clade, comprising only as-yet-uncultured phylotypes. Bacteria of this cluster occur from temperate to polar regions with highest abundance in the Southern Ocean, but not in tropical and subtropical regions. Between the south Atlantic subtropical front and Antarctica, we detected two distinct phylotypes, one north and one south of the polar front, indicating that two adjacent but different oceanic provinces allow the persistence of distinct but closely related phylotypes. These results suggest that the global distribution of major marine bacterioplankton components is related to oceanic water masses and controlled by their environmental and biogeochemical properties. PMID- 14749833 TI - Species-specific calls evoke asymmetric activity in the monkey's temporal poles. AB - It has often been proposed that the vocal calls of monkeys are precursors of human speech, in part because they provide critical information to other members of the species who rely on them for survival and social interactions. Both behavioural and lesion studies suggest that monkeys, like humans, use the auditory system of the left hemisphere preferentially to process vocalizations. To investigate the pattern of neural activity that might underlie this particular form of functional asymmetry in monkeys, we measured local cerebral metabolic activity while the animals listened passively to species-specific calls compared with a variety of other classes of sound. Within the superior temporal gyrus, significantly greater metabolic activity occurred on the left side than on the right, only in the region of the temporal pole and only in response to monkey calls. This functional asymmetry was absent when these regions were separated by forebrain commissurotomy, suggesting that the perception of vocalizations elicits concurrent interhemispheric interactions that focus the auditory processing within a specialized area of one hemisphere. PMID- 14749834 TI - SOL-1 is a CUB-domain protein required for GLR-1 glutamate receptor function in C. elegans. AB - Ionotropic glutamate receptors (iGluRs) mediate most excitatory synaptic signalling between neurons. Binding of the neurotransmitter glutamate causes a conformational change in these receptors that gates open a transmembrane pore through which ions can pass. The gating of iGluRs is crucially dependent on a conserved amino acid that was first identified in the 'lurcher' ataxic mouse. Through a screen for modifiers of iGluR function in a transgenic strain of Caenorhabditis elegans expressing a GLR-1 subunit containing the lurcher mutation, we identify suppressor of lurcher (sol-1). This gene encodes a transmembrane protein that is predicted to contain four extracellular beta-barrel forming domains known as CUB domains. SOL-1 and GLR-1 are colocalized at the cell surface and can be co-immunoprecipitated. By recording from neurons expressing GLR-1, we show that SOL-1 is an accessory protein that is selectively required for glutamate-gated currents. We propose that SOL-1 participates in the gating of non-NMDA (N-methyl-D-aspartate) iGluRs, thereby providing a previously unknown mechanism of regulation for this important class of neurotransmitter receptor. PMID- 14749835 TI - The RickA protein of Rickettsia conorii activates the Arp2/3 complex. AB - Actin polymerization, the main driving force for cell locomotion, is also used by the bacteria Listeria and Shigella and vaccinia virus for intracellular and intercellular movements. Seminal studies have shown the key function of the Arp2/3 complex in nucleating actin and generating a branched array of actin filaments during membrane extension and pathogen movement. Arp2/3 requires activation by proteins such as the WASP-family proteins or ActA of Listeria. We previously reported that actin tails of Rickettsia conorii, another intracellular bacterium, unlike those of Listeria, Shigella or vaccinia, are made of long unbranched actin filaments apparently devoid of Arp2/3 (ref. 4). Here we identify a R. conorii surface protein, RickA, that activates Arp2/3 in vitro, although less efficiently than ActA. In infected cells, Arp2/3 is detected on the rickettsial surface but not in actin tails. When expressed in mammalian cells and targeted to the membrane, RickA induces filopodia. Thus RickA-induced actin polymerization, by generating long actin filaments reminiscent of those present in filopodia, has potential as a tool for studying filopodia formation. PMID- 14749836 TI - The ADP/ATP translocator is not essential for the mitochondrial permeability transition pore. AB - A sudden increase in permeability of the inner mitochondrial membrane, the so called mitochondrial permeability transition, is a common feature of apoptosis and is mediated by the mitochondrial permeability transition pore (mtPTP). It is thought that the mtPTP is a protein complex formed by the voltage-dependent anion channel, members of the pro- and anti-apoptotic BAX-BCL2 protein family, cyclophilin D, and the adenine nucleotide (ADP/ATP) translocators (ANTs). The latter exchange mitochondrial ATP for cytosolic ADP and have been implicated in cell death. To investigate the role of the ANTs in the mtPTP, we genetically inactivated the two isoforms of ANT in mouse liver and analysed mtPTP activation in isolated mitochondria and the induction of cell death in hepatocytes. Mitochondria lacking ANT could still be induced to undergo permeability transition, resulting in release of cytochrome c. However, more Ca2+ than usual was required to activate the mtPTP, and the pore could no longer be regulated by ANT ligands. Moreover, hepatocytes without ANT remained competent to respond to various initiators of cell death. Therefore, ANTs are non-essential structural components of the mtPTP, although they do contribute to its regulation. PMID- 14749837 TI - Mechanically driven ATP synthesis by F1-ATPase. AB - ATP, the main biological energy currency, is synthesized from ADP and inorganic phosphate by ATP synthase in an energy-requiring reaction. The F1 portion of ATP synthase, also known as F1-ATPase, functions as a rotary molecular motor: in vitro its gamma-subunit rotates against the surrounding alpha3beta3 subunits, hydrolysing ATP in three separate catalytic sites on the beta-subunits. It is widely believed that reverse rotation of the gamma-subunit, driven by proton flow through the associated F(o) portion of ATP synthase, leads to ATP synthesis in biological systems. Here we present direct evidence for the chemical synthesis of ATP driven by mechanical energy. We attached a magnetic bead to the gamma-subunit of isolated F1 on a glass surface, and rotated the bead using electrical magnets. Rotation in the appropriate direction resulted in the appearance of ATP in the medium as detected by the luciferase-luciferin reaction. This shows that a vectorial force (torque) working at one particular point on a protein machine can influence a chemical reaction occurring in physically remote catalytic sites, driving the reaction far from equilibrium. PMID- 14749838 TI - Mapping life-science skills. PMID- 14749840 TI - Scientists and societies. Junior Faculty at the Karolinska Institute. PMID- 14749843 TI - The treatment of atopic dermatitis and other dermatoses with leukotriene antagonists. AB - Atopic dermatitis (AD) is a chronically relapsing eczematous disorder of the skin that occurs in persons of all ages but is more common in children. AD is associated with other atopic diseases such as allergic rhinoconjuntivitis or bronchial asthma. Nearly 80% of children with AD eventually develop allergic rhinitis or asthma. AD can be classified as mixed (cases associated with respiratory allergies) and pure . Pure AD has intrinsic and extrinsic variants. In the extrinsic type, interleukin-4 is secreted by T-cells isolated from spontaneous lesions and skin-derived T-lymphocytes express more IL-13. Due to the different immunopathogenesis, it has been suggested that antileukotriene agents may be more successful in the treatment of the extrinsic subgroup. Leukotrienes (LTs) are a class of potent biological inflammatory mediators derived from arachidonic acid through the 5-lipoxygenase pathway. There is evidence of enhanced LT production in the pathogenesis of AD. Evidence in the literature provides a pathophysiological rationale for the use of cysLT receptor blockers in the treatment of AD. However, the exact mechanism of action of leukotriene receptor antagonists in AD is not known. In small clinical and case studies, montelukast was found to be a safe and effective alternative or steroid sparing therapy in the management of patients with atopic dermatitis. PMID- 14749842 TI - Conservation biology: fatal medicine for vultures. PMID- 14749844 TI - Chemical peels. AB - With so many new peel preparations on the market today, the dermatologist must ask himself basic questions concerning the products. The most important question is directed to the medical literature rather than the advertising or marketing campaign so common among market-driven cosmetic products. Since all peeling agents--superficial, medium depth and deep--are derived from basic chemicals known to cause exfoliation, destruction and/or inflammation of skin in a controlled manner, the clinician must ask what is new and better about the product. Peeling agents, regardless of their proprietary new name, fall into chemical families. The clinical evaluation of these generic agents is well documented in our literature as to efficacy, technical care and safety. In addition, combinations of peeling agents have been presented in the dermatologic cosmetic literature with scientific clinical trials and histology. These include: 1) The Gordon-Baker phenol peel; 2) Combination medium depth peeling; 3) Glycolic acid formulations. It is the responsibility of the dermatologic surgeon to be in control of his chemicals and his products. It is thus necessary for him to understand all the products and the peel formulation and be sure it has undergone the test of objective scientific study with clear clinical evaluations and histology. Only then will we truly know the effectiveness of the agents we are using for exfoliating and resurfacing. PMID- 14749845 TI - Predictors of the development of impaired fasting glucose versus impaired glucose tolerance are partly different in men: a 6-year follow-up study. PMID- 14749846 TI - Construction of a binary BAC library for an apomictic monosomic addition line of Beta corolliflora in sugar beet and identification of the clones derived from the alien chromosome. AB - A plant-transformation-competent binary BAC library was constructed from the genomic DNA of the chromosome 9 monosomic addition line of Beta corolliflora Zoss. in sugar beet ( B. vulgaris. L). This monosomic addition line (designated M14) is characterized by diplosporic reproduction caused by the alien chromosome carrying the gene(s) responsible for diplospory. The library consists of 49,920 clones with an average insert size of 127 kb, representing approximately 7.5 haploid genome equivalents and providing a greater than 99% probability of isolating a single-copy DNA sequence from the library. To develop the scaffold of a physical map for the alien chromosome, B. corolliflora genome-specific dispersed repetitive DNA sequences were used as probes to isolate BAC clones derived from the alien chromosome in the library. A total of 2,365 positive clones were obtained and arrayed into a sublibrary specific for B. corolliflora chromosome 9 (designated bcBAC-IX). The bcBAC-IX sublibrary was further screened with a subtractive cDNA pool generated from the ovules of M14 and the floral buds of B. vulgaris by the suppression subtractive hybridization method. One hundred and three positive binary BACs were obtained, which potentially contain the genes of the alien chromosome specifically expressed during the ovule and embryo development of M14, and may be associated with apomictic reproduction. Thus, these binary BAC clones will be useful for identification of the genes for apomixis by genetic transformation. PMID- 14749847 TI - Molecular markers derived from RAPD, SCAR, and the conserved 18S rDNA sequences for classification and identification in Pyrus pyrifolia and P. communis. AB - We generated RAPD, SCAR, and conserved 18S rDNA markers for classifying and identifying cultivars of Pyrus pyrifolia (Japanese pear) and P. communis (European pear). PCR amplification with selected specific primers-LCH327UP and LCH327DOWN-was performed using DNA extracted from 25 P. pyrifolia and P. communis cultivars. The 1,380-bp fragment was amplified from P. communis cvs. Beurre Giffard, Cascade, Conference, Clapp's Favorite, Packhams Triumph, and Winter Nelis. RAPD has only a dominant single band of 1,380-bp, however, SCAR has one or more band of the same size. Amplification involving sequence-specific primer pairs LCH346UP and LCH346DOWN resulted in a loss of polymorphism. The 1,190-bp fragment was amplified from all P. pyrifolia cultivars. The conserved sequences of the 18S rDNA fragment of 25 pear cultivars were amplified and analyzed with 42 restriction enzymes. Compared with P. pyrifolia cultivars, they lacked the restriction enzyme site of KpnI and had one less RsaI site. Cultivar Gamcheonbae had a specific PstI restriction site, while cvs. Mansoo and Conference pear digested with AluI showed a different presentation than other cultivars. For the Okusankichi and Shinil pears TaqI was best marker for identification in P. pyrifolia. These results can be adopted for identifying pear cultivars; to date there is no standard marker for identifying the cultivars of fruit trees in Korean fruit tree breeding programs. PMID- 14749848 TI - [Difficulties involved in the Hoffa fractures]. AB - To study the outcome in Hoffa fractures regarding the type of fracture, concomitant injuries, and different treatments. From January 1996 until December 2002, 7 Hoffa fractures (6 patients) from a total of 79 distal femoral fractures were treated in the Department of Trauma Surgery, University Hospital of Giessen. The results were evaluated by Neer's score. For the evaluation of postoperative vascularization of the distal femur, clinical and radiological findings were compared with anatomical studies of the bone arteries by corrosion preparations. Fracture consolidation was seen in all cases. Patients with an isolated Hoffa fracture achieved a score of 89.0 (82-97) according to Neer. One patient suffering initially from nonunion achieved 68 points after bone union. One polytraumatized patient with bilateral Hoffa fracture achieved a satisfying result (score value of 72) and a bad result (score value of 38). Multiple small nutrient arteries of the distal femur originate from the "rete articulare genus." They can be expected to ensure the arterial blood supply. Hoffa fracture is a very rare injury. In our patients, careful surgical treatment with open reduction and safe screw fixation yielded good to very good results. PMID- 14749849 TI - [Trauma due to blank cartridges and fireworks. Early and correct treatment prevents tattooing and scarring]. AB - Traumata secondary to blank cartridges and fireworks are not harmless at all. They can cause cosmetic disfigurement due to permanent tattooing and scars. Often the face and hands are injured. Fourteen patients with blast injuries were treated between 1992 and 2002 in our clinic. The average age was 20.4 years (range: 13-41 years, median: 17.5 years). Eight patients were aged under 18 years. Most of the victims were males (11 of 14). According to the extent of the powder tattooing we removed the particles under local or general anesthesia within 24 h. We used sterile tooth and hand brushes. After the operation we treated the wounds with local antibiotics. We describe two cases to illustrate our procedure. Early correct treatment of tattooing caused by fireworks or gun powder explosions within 24 h up to a maximum of 72 h prevents the development of permanent cosmetic disfigurement. The technique is simple, effective, and saves the cost of later removal of traumatic tattoos by laser or dermabrasion. PMID- 14749850 TI - [Microfracture technique for treatment of osteochondral and degenerative chondral lesions of the talus. 2-year results of a prospective study]. AB - The microfracture technique has become an established method for the treatment of chondral lesions in the knee. The aim of this study was to determine its usefulness in the treatment of osteochondral (OCL) and degenerative chondral lesions (DCL) of the talus. In a prospective study, 32 patients with an osteochondral lesion (22 patients) or degenerative chondral lesion (10 patients) were operated arthroscopically and treated with the microfracture technique. All patients were evaluated preoperatively as well as 3, 6, 12, and 24 months postoperatively with the Hannover scoring system and a visual analog score (VAS). MRI scans were performed at all points. Statistical analyses were performed using the Wilcoxon sign rank-sum test with a level of significance of p < or = 0.05. At a mean follow-up of 2 years (range: 22-27 months), 23 patients (14 OCL, 9 DCL) were available for follow-up. The average age of the 17 male and 15 female patients at the time of operation was 39 years (range: 16-74 years). The results according to the Hannover scoring system were 43% excellent, 35% good, and 22% satisfactory with 93% excellent or good results for OCL. Older patients (>50 years, n=8) had slightly better results than younger patients. VAS revealed an average of 7.84 (preoperative: 3.53) for pain, 7.85 for function (preoperative; 3.38), and 7.78 for satisfaction (preoperative: 2.41). MRI and arthroscopic assessment showed a visible regeneration of the cartilage in the microfractured area. In a short-term follow-up, the microfracture technique has proven to restore severe cartilage damage with a good functional outcome. Age is not a limiting factor. However, longer-term results remain to be evaluated. PMID- 14749851 TI - [Restoration of the ability to walk through bilateral triple arthrodesis in marked spastic pes equinovarus]. AB - A tendon transfer is the method of choice in easily reducible pes equinovarus. However, in long-time persisting deformities with spasms, a plantigrade position can not be maintained with these procedures. Therefore, we perform an additional bilateral triple arthrodesis in a patients with such bilateral deformities. A 55 year-old woman developed, within the scope of several surgical procedures on the cervical spine, marked bilateral pes equinovarus and flexion contractures of the knees. The patient's ambulation was limited to a wheelchair for 3 years. Then, in an interval of 1 year, we performed an unilateral soft tissue release, z-tenotomy of the Achilles tendon, triple arthrodesis with correction of the deformity, and posterior tibial tenden transfer. At follow-up 5 years after the second procedure, the 61-year-old patient was able to walk alone with two walking sticks. In the case described, the correction of a marked pes equinovarus with spasms, which was achieved by an extensive soft tissue release, could be stabilized through a triple arthrodesis in such way that the plantigrade position of the foot could be controlled through a posterior tibial tendon transfer. PMID- 14749852 TI - [Benign desmoplastic fibroblastoma of the lower leg. A rare case]. AB - Desmoplastic fibroblastoma ("collagenous fibroma") is a benign fibrous soft tissue tumor. We report a rare case of a "collagenous fibroma" located in the proximal lower leg in 56-year-old man. The clinical, radiological, and pathological-morphological findings are documented. The operative treatment was performed according to general principles of surgical management of benign soft tissue tumors. PMID- 14749853 TI - [Surgical indications in intraosseous lipoma of the calcaneus. Case report and critical review of the literature]. AB - Intraosseous lipomas represent a small number of benign bone tumors with incidence rates of approximately 0.1%. In about 15% these neoplasias are localized within the calcaneus, mostly at Ward's triangle. The tumors usually remain clinically inapparent and diagnosis is often obtained incidentally. Although CT scan and MRI provide specific and sensitive diagnostic tools that can distinguish morphology and dignity, surgical treatment is not standardized yet. In conjunction with a case report, we summarize and critically compare current treatment strategies. PMID- 14749854 TI - [Hyperparathyroidism -- a differential diagnosis of cystic bone tumors]. AB - Clinical manifestations of primary hyperparathyroidism include neuromuscular, intestinal, and osseous symptoms with osteolytic lesions of the bone and pathological fractures. In most cases a primary adenoma of the parathyroid gland leads to an increased production of the parathormone with subsequent osteoclastic bone catabolism. Surgical treatment entails removal of the parathyroid adenoma. We present a female patient and the possible diagnostic and differential diagnostic difficulties encountered in interdisciplinary management. PMID- 14749856 TI - [New dermatologist's procedure. Research plans for improving secondary prevention of occupational dermatoses]. AB - The dermatologist's procedure was introduced in 1972 by employers' liability insurance funds in the industrial, agricultural, and public sectors of Germany's statutory occupational accident insurance as a "procedure for early detection of occupational skin diseases." So far, it is still the most relevant tool for secondary prevention in occupational dermatology in Germany. According to the intention of this procedure, insured persons with a skin disease in which an occupational etiology is suspected must be offered preventive measures and, if necessary, given appropriate treatment to avoid job-loss. To improve the efficiency of the "classic" dermatologist's procedure, in 1999, on the initiative of the Central Federation of Industrial Professional Associations, a study group was founded in cooperation with the Task Force on Occupational and Environmental Dermatology of the German Dermatological Society and the Professional Organisation of German Dermatologists. In October 2002, a controlled intervention study started in North-western Germany to scientifically evaluate the "optimised" dermatologist's procedure, which was proposed by the study group and compare it to the standard ("classic") procedure. The study results should allow detailed suggestions for an improved dermatologist's procedure before it is introduced nationwide. PMID- 14749857 TI - [Skin cancer and occupational disease]. AB - Although it is universally accepted that UV light exposure can cause malignant skin tumors, UV-induced skin cancers are not recognized as an occupational disease in Germany. Exposure to natural or artificial UV light occurs in many work places, so that the induction of occupational skin cancers is certainly plausible. In recent years, a special clause in the occupational disability rules has recognized some cases of UV-induced skin cancers. We discuss the nature of occupational UV exposure, explore preventative measures and review the data regarding occupational UV-induced skin tumors. After evaluating recent publications, we conclude that for squamous cell carcinoma the epidemiological proof of an at least doubled risk (RR >2) due to occupational UV radiation can be given. The clear dose response relationship supports these epidemiological findings. For the individual risk assessment, an attributive UV radiation >40% due to occupational factors must exist. Under those circumstances, squamous cell carcinoma should be recognized and compensated as an occupational disease. PMID- 14749855 TI - [DRG reimbursement for multiple trauma patients -- a comparison with the comprehensive hospital costs using the German trauma registry]. AB - The introduction of diagnosis related groups (DRG) will radically change the payment system for German hospitals. In 2002 the values for most DRG's were published for the german system (G-DRG). The polytrauma working group of the German Trauma Society developed a calculating algorithm to estimate the comprehensive hospital costs for every patient in the German trauma registry. The aim of this study was to compare these costs with the reimbursement according the the G-DRG's for a standardized population of polytrauma patients. MATERIAL AND METHODS: For polytrauma patients treated at Hannover Medical School in 2000 and 2001 the reimbursement according to the G-DRG's was calculated using a base value of 2900 euro. In the same patients the total cost of inpatient treatment was calculated according to the algorithm developed by the polytrauma working group of the German Trauma Society. The difference between these values represents the economic result. This was calculated as an overall result, but also for specific subgroups of patients (injury severity, mortality, G-DRG grouping). RESULTS: Datasets of 103 polytrauma patients were included. The following G-DRG's were most frequently occuring: A06Z (n=41), A07Z (n=16), W01Z (n=13). All other G DRG's were documented less than 3 times. The mean reimbursement according to the G-DRG was 21.380+/-12.300 euro for a polytrauma patient. However, the mean hospital cost accounted to 34.274+/-22.501 euro, which resulted in a mean deficit of 12.893+/-15.534 euro. Analysis of subgroups revealed, that an ISS of more than 35 points, patients with a prolonged hospital stay and patients of the G-DRG group A06Z show a particularly negative result. CONCLUSION: The comprehensive hospital costs for treating polytrauma patients are on average 12.893 euro higher than the reimbursement according the G-DRG's. For hospitals to be fully reimbursed G-DRG values have to be reconfigured according to the German health care system. Thus, inclusion criteria to specific G-DRG have to be changed and a specific G-DRG group for very severely injured patients needs to be established. PMID- 14749858 TI - [Quality of life of patients with occupationally-induced hand eczema]. AB - In recent years quality of life has been studied in a growing number of different dermatological diseases. Internationally validated questionnaires such the RAND 36 (identical to the SF-36) do not contain enough questions which are relevant for skin diseases. There is no publication on quality of life issues in occupational skin diseases, and only one short report gives data on quality of life in hand eczema. The widely used skin specific instrument DLQI has only 2 questions that indirectly refer to employment issues. A quality of life questionnaire on occupational skin diseases (mostly hand eczema) should ideally include questions on work-related impairment of both physical functioning and interaction with colleagues. PMID- 14749859 TI - [Allergic occupational dermatoses. Role of in vitro studies in diagnosis and prevention]. AB - Recently published guidelines only recommended in vitro assays for atopy screening by the determination of specific IgE antibodies in occupational cutaneous allergic diseases. However further indications include latex allergy, occupational drug allergy and wasp/ bee venom allergy. The lymphocyte transformation test may be an alternative to the patch test, in order to differentiate allergy from irritancy, or it may be an alternative to the patch test if hazardous compounds such as carcinogens are the possible antigens. PMID- 14749860 TI - [Dermatologic occupationally relevant type I allergies]. AB - In Germany the most frequent initial report of occupational disease is due to occupational skin diseases ("Berufskrankheitenanzeige nach Nr. 5101 der Berufskrankheitenverordnung") defined as "severe or recurrent skin diseases that force the discontinuation of any activity that causes or that could be causing the development, the worsening, or the recurrence of the skin disease". The majority of these occupational skin diseases consists of irritant and allergic contact dermatitis. In contrast, work place- related Type I allergy (contact urticaria syndrome) is less frequent, but carries the risk of systemic reactions. The clinical manifestations and pattern of exposure to Type I allergens in the work place are described. PMID- 14749861 TI - [Internet and occupational dermatology. Quality criteria and useful links]. AB - The World Wide Web (WWW) offers physicians as well as their patients a huge resource of health information, which can be attained within minutes. Nevertheless it is difficult to select the needed information out of the mass of provided data and their quality is often poor with many unproven statements. Occupational dermatologists can especially benefit from the internet as they need a wide range of data from many sources. Much of this data can already be obtained via internet. To facilitate their search for information, this article presents a selection of German websites. The quality of the websites has been assessed using criteria such as design, search function, provided links and medical information. The results revealed deficiencies in the transparency of the data, e.g. often it is not apparent who is responsible for the information or when they were updated. PMID- 14749862 TI - [The excimer laser in dermatology and esthetic medicine]. AB - First reports about the use of the excimer laser in dermatology date back to 1997. It is seen as an improvement on conventional phototherapy and photochemotherapy because of the lower cumulative UV-dose involved, the shorter time frame required for treatment and the option of targeting individual lesions without affecting the surrounding healthy skin. In addition to the indications of psoriasis vulgaris, vitiligo and atopic eczema (for which there is now FDA approval in the US), the spectrum of possible uses for the excimer laser is growing rapidly, especially in the field of light-sensitive dermatoses. Case studies so far have ranged from post-operative hypopigmentation to acne vulgaris and from alopecia areata to parapsoriasis en plaque. The foremost priorities in the future will be to evaluate reproducible therapeutic regimens with realistic prospects of success in large-scale studies; assess potential iatrogenic risks in treatment; develop pathogenetic models for the mechanism of action; and define therapeutic approaches to new indications. This paper summarizes the publications to date and discusses our observations and experiences. PMID- 14749864 TI - [Unilateral facial edematous granulomatosis. Initial symptom of basalioma]. AB - A seventy-year-old woman presented with persistent edema of the left lower eyelid and induration in the inner angle of her left eye. Histological examination revealed granulomatous blepharitis of the lower eyelid as well as a solid basal cell carcinoma with partial calcification in the inner angle of the eye. The edema of the lower eyelid resolved upon complete surgical removal of the tumor. PMID- 14749863 TI - [Topically applied arginine hydrochloride. Effect on urea content of stratum corneum and skin hydration in atopic eczema and skin aging]. AB - BACKGROUND AND OBJECTIVE: Currently, there are no data on how the topical application of amino acids influences the complex moisture retaining system of the skin in vivo. PATIENTS/METHODS: An open study was performed to investigate the effects of topical application of arginine hydrochloride on epidermal stratum corneum urea content, transepidermal water loss, skin hydration, and clinical status of patients with atopic dermatitis and dry elderly skin. RESULTS: Treatment of patients with atopic dermatitis with 2.5% arginine hydrochloride ointment over 4 weeks showed a significant increase in urea in the stratum corneum as well as a continuous increase in skin moisture. CONCLUSIONS: The urea deficit in the stratum corneum in atopic dermatitis and elderly skin was corrected not by applying the moisturizer urea itself but instead by using arginine - its precursor in the Krebs-Henseleit urea cycle. This topical treatment also improved the clinical symptoms of dry skin. PMID- 14749866 TI - [Disseminated eruptive syringomas in Down syndrome]. AB - A 42-year-old male with Down's syndrome suddenly developed disseminated red-brown papules. Histological studies revealed eruptive syringomas. We discuss the association of Down syndrome with different skin diseases, in particular with syringomas. PMID- 14749865 TI - [Adiponecrosis subcutanea neonatorum in congenital pneumonia]. AB - In the course of a connatal pneumonia, a 7-day-old female newborn developed symmetrical subcutaneous nodules on her back, shoulders, and upper arms. These skin lesions were accompanied by hypercalcemia. Histological examination confirmed the putative clinical diagnosis of subcutaneous fat necrosis of the newborn. We discuss the differential diagnoses, therapeutic strategies, and prognosis of this uncommon disorder of the fat tissue. PMID- 14749867 TI - [Unilateral cheek tumor. Case report and differential diagnoses]. AB - A 56-year-old male presented with a tumor on the cheek that had developed over 5 months. Unexpectedly, histological and immunohistochemical studies revealed an adenocarcinoma of the salivary gland. We discuss the differential diagnoses of tumors of the cheek. PMID- 14749868 TI - [Folliculitis barbae in herpes simplex infection]. AB - A 60-year-old male athlete developed a folliculitis in the beard region after several competitions. After identification of herpes simplex antigen within the lesions, systemic therapy with acyclovir led to rapid improvement. In folliculitis resistant to antibiotic and anti-inflammatory therapy, viral and mycotic infections as well as eosinophilic folliculitis should be considered as differential diagnostic possibilities. PMID- 14749869 TI - [Cutaneous infection with Mycobacterium marinum. successful therapy with rifampicin and clarithromycin]. AB - We report on two male patients with cutaneous Mycobacterium marinum infection. After the diagnosis was established by culture, therapy with rifampicin and clarithromycin over a period of three months led to remarkable improvement. PMID- 14749870 TI - [Type I sensitization to seminal fluid. Urticaria and angioedema]. AB - Allergy to human seminal fluid is a rare but potentially life-threatening disorder. Acute symptoms range from local reactions to generalized symptoms. A 30 year-old woman experienced vulvovaginal itching, erythema and swelling as well as generalized urticaria and angioedema of the neck and face after unprotected sexual intercourse. A scratch test with undiluted seminal fluid and the detection of specific IgE antibodies revealed a type I allergy to seminal fluid. Risk factors and therapeutic options are discussed. PMID- 14749871 TI - [Syphilis. Clinical aspects of Treponema pallidum infection]. AB - Syphilis is a sexually transmitted infection by Treponema pallidum. Without antibiotic treatment syphilis lasts for several decades and may develop up to 4 different clinical stages. Usually, the disease begins with a distinct painless and indurated ulcer at the contact site: the primary chancre. An indolent regional lymph node swelling is usually associated with the syphilitic chancre. After spontaneous healing of the primary lesion and several weeks of latency, the clinical symptoms of secondary syphilis occur. Treponema pallidum bacteremia leads to common symptoms like fever and malaise, but also to a generalized lymphadenopathy, and a broad variety of lesions of the skin and mucosal membranes. Non-pruritic transient exanthems often involving palms and soles, condylomata lata, and a specific angina with mucous patches of the oral cavity are prominent signs. After several relapses, which are characterized by a decreasing intensity of clinical symptoms, secondary syphilis then resolves spontaneously. A second period of latency follows, lasting 3-12 years. Then the outcome of untreated syphilis becomes apparent: spontaneous healing by elimination/inactivation of the spirochetes (75%) or transition to tertiary syphilis (25%). Two kinds of granulomatous skin reactions are typical for tertiary syphilis: superficial nodular syphilids and gummas. The bones, as well as the cardiovascular and central nervous system, may also be involved. Finally, metasyphilis with severe and sometimes lethal neurological symptoms (tabes dorsalis, progressive paralysis) occurs 10 to 30 years after primary infection. Except for irreversible tissue destruction which occurs prior to therapy, all stages of syphilis can be cured completely. PMID- 14749873 TI - [The effects of active and passive humidification on ventilation-associated nosocomial pneumonia]. AB - STUDY OBJECTIVE: Airway humidification of ventilated patients in an intensive care unit may be established by heated humidifying systems (active) or by the means of a (passive) heat and moisture exchange filter (HMEF). There is a controversial discussion about the influence of the type of humidification on the rate of ventilator-associated pneumonia (VAP). Among 3,585 patients both methods were tested over a period of 21 months in an open, non-randomized cohort study. The aim of the investigation was to compare the incidence of VAP caused by a change of humidification strategy. METHOD: All patients in a 16-bed surgical intensive care unit who required mechanical ventilation, were included. In the first period (period AB) 1,887 cases were handled with a heated humidifier. During the second period (period PB) 1,698 patients were treated using a HMEF. Infection control was established according to the national Infection Surveillance Program (KISS) based on the CDC criteria for VAP. RESULTS: During the period of 42 months, 99 cases of VAP were reported. The incidence for VAP was found to be 13.5 (AB) and 9.6 (PB) per 1,000 ventilator days, a rate of 32.3 and 22.4 VAP per 1,000 patients, respectively. The rate of VAP among the groups ( p=0.068) and the incidence of VAP per 1,000 ventilator days ( p=0.089) only just failed to reach a significant level, but in the group of patients requiring mechanical ventilation for more than 2 days, the difference did reach statistical significance ( p=0.012). CONCLUSION: Our results showed that the rate of VAP could be significantly reduced by changing the strategy from active to passive humidification devices, especially concerning patients requiring long-term respirator therapy. A more physiological humidification and a reduced number of airway manipulations are discussed as a possible explanation. PMID- 14749874 TI - [Bispectral index and desflurane concentration below 1 MAC]. AB - INTRODUCTION: We investigated the relationship between bispectral index (BIS, A 2000, Aspect Medical Systems, USA) and end-tidal desflurane concentrations below 1 MAC which is especially the range of interest if desflurane is combined with remifentanil for fast-track anaesthesia. METHODS: With institutional review board approval and written informed consent we investigated 50 adult ASA I-III patients scheduled for minor orthopaedic surgical procedures. The BIS electrode (BIS Sensor, Aspect Medical Systems, USA) was positioned on the patient's forehead as recommended by the manufacturer. All patients were premedicated with 0.15 mg x kg(-1) diazepam orally in the evening and on the morning before surgery. Induction of anaesthesia was started with a remifentanil infusion at 0.4 micro g x kg(-1) x min(-1) and 5 min later 2 mg x kg(-1) propofol was given for hypnosis. Immediately after intubation, remifentanil was reduced to 0.2 micro g x kg(-1) x min(-1), and the depth of anaesthesia was adjusted according to clinical needs by regulating the desflurane concentration. Starting 20 min after induction, a total of 1,483 data pairs, i.e. end-tidal desflurane concentrations with corresponding BIS values, was recorded. During surgery data pairs were analysed by linear regression analysis for each patient separately. During emergence from anaesthesia, desflurane effect compartment concentrations were simulated and non linear regression analysis was applied. RESULTS: We found a relationship between BIS ranges and mean end-tidal desflurane concentrations: A BIS range of 100-85 was related to 1.1+/-0.7 vol%, BIS 84-65 to 1.5+/-1.2 vol%, BIS 64-50 to 2.9+/ 1.3 vol% and BIS 49-40 to 3.1+/-0.9 vol%. In particular, end-tidal desflurane concentrations related to these BIS ranges were significantly different from each other (ANOVA; P<0.05). Intraoperatively, we obtained a correlation coefficient of R=0.42+/-0.24 (mean+/-SD, range 0.05-0.96), during emergence from anaesthesia the correlation coefficient was R=0.84+/-0.12 (range 0.61-0.99). CONCLUSION: We could demonstrate a relationship between bispectral index and end-tidal desflurane concentrations below 1 MAC with a BIS range of 49-40 being associated with approximately 0.5 MAC of desflurane during desflurane-remifentanil-anaesthesia. PMID- 14749872 TI - [Role of the innate immune response in sepsis]. AB - The innate immune system succeeds against the majority of infections before the adaptive immune system is activated. New findings contribute to a better understanding of the pathophysiology of sepsis and lead to the development of new therapeutic strategies. The innate immune system, being responsible for the first response to infections, can trigger adaptive immune responses in case the initial response is ineffective. Both arms of the immune system interact with each other, mainly via cell-cell-interactions but also by soluble factors, such as cytokines and chemokines. Two sub-populations of helper T-cells direct both balanced activation and inhibition of the two arms of the immune systems using specific patterns of cytokine release. Results obtained in new animal models of sepsis, taking a progressive growth of bacteria into account, have implied that existing knowledge has to be reanalyzed. The idea of sepsis as a mere "over-reaction to inflammation" has to be abandoned. Various so-called pattern recognition receptors (e.g. toll-like receptors, TLRs, NOD proteins) are located intracellularly or in the plasma membrane of innate immune cells and recognize certain patterns expressed exclusively by extracellular pathogens. Upon receptor engagement, intracellular signaling pathways lead to cellular activation, followed by release of various cytokines and anti-microbial substances. During the course of sepsis a cytokine shift towards increasing immune suppression occurs. The innate immune system also contributes to the migration of leukocytes in inflammed tissue, involving chemokines and adhesion molecules. Leukocytes also secrete the tissue factor leading to formation of thrombin. The environment in sepsis can cause disseminated intravascular coagulation (DIC), but at the same time thrombin triggers the release of chemokines and adhesion molecules through endothelial cells, which represents a positive feedback mechanism for innate immune responses. New therapeutic strategies for sepsis try to establish a well balanced immune response. Intervention is accomplished through inhibition of inflammatory cytokines, their receptors or through activation of immunostimulatory responses. PMID- 14749875 TI - [Intracerebral hemorrhage after cesarean section under spinal anesthesia. Coincidence or causality?]. AB - Intracerebral haemorrhage is a rare complication of spinal anaesthesia in obstetrics. A 37-year-old woman without any accompanying disease during a twin pregnancy, underwent an urgent caesarean section due to insufficiency of the placenta under spinal anaesthesia using hyperbaric bupivacain (0.5%) and a pencil point spinal needle Sprotte 27 Gauge. The patient developed severe headache, a hemiparesis of the right upper limb and became somnolent and finally unconscious 80 min after the procedure. An immediately performed computed tomographic scan revealed a large acute intracerebral haemorrhage in the left hemisphere region with mass effect. The patient underwent temporoparietal craniotomy. No obvious cause of the haemorrhage, such as aneurysm or arteriovenous malformation was found. The patient fully recovered and was weaned from the respirator 32 h postoperatively. On postoperative day 7 the remaining neurologic deficits included aphasis and severe hemiparesis of the right upper limb and a right extensor plantar response. The neurologic status did not improve substantially until 6 months after the complication. The case and the recent literature are discussed. PMID- 14749876 TI - [Spontaneous intracranial hypotension. A rare syndrome with good treatment options]. AB - The spontaneous intracranial hypotension syndrome is a rare event but with increasing tendency. The clinical characteristics are comparable to those occurring after dural puncture and the most important clinical finding is the postural headache. The syndrome results from cerebrospinal fluid leakage but its etiology is still nearly unknown. The leaks are mainly located cervically or at the cervicothoracic junction. The syndrome may be associated with cranial subdural fluid build-up. Magnetic resonance imaging of the brain typically reveals diffuse pachymeningeal enhancement, frequently in association with displacement of the brain. Knowledge of this can be helpful to facilitate the diagnosis. Although conservative measures are often initially undertaken, placement of an epidural blood patch is the treatment of choice. Because of its similarity to postdural puncture headache, anaesthesiologists and pain therapists are increasingly involved in diagnosis and therapy. We report 2 patients with spontaneous intracranial hypotension. In addition to the cardinal feature of a postural headache, the patients suffered from subdural fluid build-up demonstrated by cranial magnetic resonance imaging. PMID- 14749877 TI - [Hypotensive cardio-circulatory failure and metabolic acidosis after suicidal intoxication with trimipramine and quetiapine. Case report and background]. AB - The case of a 44-year-old female patient is reported, who ingested trimipramine and quetiapine in a suicide attempt. Initially sinus tachycardia and hypotension were seen, which resulted in a hypotensive cardio-circulatory failure despite fluid therapy and administration of catecholamines. Because of the life threatening situation and the fact that the ingestion was 2 h prior to admission, a rapid transport to the next hospital was preferred to treatment with active charcoal. Intoxication with tricyclic antidepressants are very common in Europe and have a mortality of up to 15% in severe cases. The specific therapy consists of airway management, hemodynamic stabilization and primary elimination of the poison. Secondary detoxication is less important. The administration of the antidote physostigmine is controversial but carbo medicinalis should be given orally or via a gastric tube. PMID- 14749878 TI - [Apoptosis as a pathomechanism in sepsis]. AB - Sepsis is still a leading cause of death in many intensive care patients. The pathophysiology of the disease is dominated by complex immune cascades. Recent research demonstrates that immune cells respond to sepsis with an increased rate of programmed cell death. Up-regulated apoptosis of leukocytes was observed in animal models of sepsis as well as in patients suffering from severe sepsis. The mitochondrial protein Bcl-2 and the caspase cascade play an important role in the regulation of apoptosis. Overexpression of Bcl-2 or inhibition of caspases resulted in an increased survival in animal models of sepsis. Recent reports indicate the relevance of apoptosis in patients with severe sepsis. These results may spawn novel immunomodulatory strategies in the treatment of sepsis. PMID- 14749879 TI - [Feedback control of muscle relaxation with a varying on-off controller using cisatracurium]. AB - BACKGROUND: Under clinical conditions constant neuromuscular blockade can also be maintained by a simple closed-loop system. However, delayed onset time, non linearity of the dose-response curve and different sensitivity to muscle relaxants for each patient are limiting factors. METHODS: In 20 patients who underwent elective surgical procedures under continuous propofol/alfentanil anaesthesia and relaxation with cisatracurium, the maintenance of an electromyographically controlled cisatracurium block of 90% was achieved by a varying on-off control system. Using an own computer-aided measuring device, the course of the neuromuscular blockade and deviations from the desired neuromuscular block were registered. RESULTS: Over a period of 64.2+/-14.0 min, neuromuscular block could be controlled on average at a T(1)-level of 10% (90% block). The mean error of the deviation of the obtained neuromuscular blockade from the set-point was -1.6+/-0.9% on average. To maintain this neuromuscular blockade, a dose rate of 1.4+/-0.9 micro g x kg(-1) x min(-1) cisatracurium was necessary. CONCLUSIONS: It can be concluded that a simple closed-loop system allows the safe use of the intermediate term muscle relaxant cisatracurium for the performance of surgical procedures. PMID- 14749881 TI - [Legal aspects of medical continuing education]. PMID- 14749880 TI - [Microcuff pediatric tracheal tube. A new tracheal tube with a high volume-low pressure cuff for children]. AB - BACKGROUND: Principles and characteristics of the recently introduced Microcuff paediatric tracheal tube (Microcuff, GmbH, Weinheim, Germany) with anatomically based depth markings, cuff-free subglottic tube shaft and short high volume-low pressure cuff with ultrathin cuff membrane are presented. First available tubes (ID 4.0 mm) were evaluated regarding cuff pressures required to seal the trachea and regarding the distance from the tube tip to the carina. METHODS: After obtaining approval of the local ethical committee, 20 children aged 2-4 years, receiving tracheal intubation under general anaesthesia with muscle paralysis, were included. The tubes were placed during direct laryngoscopy and the glottic depth marking placed between the vocal cords. Cuff pressure to prevent audible air leakage at standardised ventilator settings (PIP 20 cm H(2)O/PEEP 5 cm H(2)O/RR20 x min(-1)) was assessed by means of a cuff pressure manometer within 5 min after intubation. Subsequently, the distance from the tube tip to the tracheal carina was measured by means of fibre bronchoscopy. Data are presented as the median (range). RESULTS: Patient age was 3.0 years (2.0-3.9 years), weight 13.5 kg (9.1-19.2 kg) and body length 95 cm (79-105 cm). The lowest cuff pressure required to seal the trachea ranged from 4-14 cmH(2)O (median 10 cm H(2)O), the distance from tube tip to tracheal carina was to 2.9 cm (2.0-4.5 cm). CONCLUSIONS: The new Microcuff paediatric tracheal tube with ultrathin high volume-low pressure cuff required tracheal sealing pressures below tracheal wall pressures usually required with uncuffed tracheal tubes for efficient sealing and ventilation at 20 cm H(2)O peak inspiratory pressure. The distance from the tube tip to carina was in the safe range in all patients. PMID- 14749882 TI - [Delaying defibrillation to give basic cardiopulmonary resuscitation to patients with out-of-hospital ventricular fibrillation]. PMID- 14749883 TI - Childbirth with labor analgesia. What is important to our patients? PMID- 14749884 TI - [Brachial plexus. Comment on the paper "Brachial plexus--anesthesia and analgesia" von S. Schulz-Stubner (Anaesthesist 2003 52:643-656)]. PMID- 14749885 TI - Biomechanics of the dental arch and incisal crowding. AB - During horizontal contact, the dental arch represents a link chain in which two convex articular surfaces are in contact and are tensioned by the dentogingival and dentoalveolar fibrous tissue. Joints composed of convex-convex surfaces are equivalent to stretched dimeric link chains whose links are in a mechanically unstable position under compression. Experiments on plaster models show that the dimensional stability of an articulated (dental) arch is considerably increased when a concave and a convex articular surface are in contact, as these joints are equivalent to an overlapping dimeric link chain whose links are in a mechanically stable position when under compression. In the abraded denture of Stone Age man, horizontal interdental contacts of transversally concave-convex dental surfaces may be found extending even into the incisal region. Increased stability of the dental arch and reduced problems of incisal crowding are to be expected if an overlapping dimeric chain is produced morphologically in each horizontal contact by means of slight interproximal enamel reduction. PMID- 14749886 TI - Effects of crowding in the lower anterior segment--a risk evaluation depending upon the degree of crowding. AB - AIM: The aim of this study was to investigate the extent to which the anterior segment is at risk of developing disease under the influence of different types of anterior crowding and whether the degree of crowding correlates with a potential risk. PATIENTS AND METHOD: A clinical examination and cast analysis taking special account of the age and gender of 125 adult patients (63 women, 62 men) were used to determine the clinical degree of abrasion and the individual incidence of tooth infractions, tooth fractures, caries, gingivitis, periodontitis and gingival recessions (WHO-OHS method), with any correlation between the incidence of disease and the respectively calculated crowding being recorded. The diagnosis of crowding was based on the segmented arch analysis proposed by Lundstrom [29]. Besides evaluating the total collective, a separate evaluation was performed for the age groups 18-34 years (n = 63) and > or = 35 years (n = 62). RESULTS: No gender-related differences in crowding were determined. Patients > or = 35 years showed significantly more crowding. An age related increase in disease was recorded. The degree of abrasion and the individual incidence of caries did not correlate with the degree of crowding. By contrast, differences in the degree of crowding were determined in the total collective for tooth infractions (p < 0.001), tooth fractures (p = 0.004), gingival bleeding (p = 0.022), shallow periodontal pockets (p < 0.001), and gingival recessions > 3.5 mm (p < 0.001). The degree of crowding was found to correlate in the younger patients with tooth infractions (p < 0.017) and tooth fractures (p = 0.036), and in the older patients with shallow periodontal pockets (p < 0.001) and gingival recessions > 3.5 mm (p < 0.001). The incidences of disease in virtually physiologic cases of crowding (crowding < or = 2 mm, n = 31) were compared with those recorded in extreme cases of crowding (crowding > or = 5 mm, n = 30). All cases of crowding > or = 5 mm were subject to gingivitis and tooth infractions, and shallow periodontal pockets occurred three times more often and gingival recession > 3.5 mm twelve times more often. The presence of deep periodontal pockets could not be attributed to the degree of crowding. CONCLUSION: With respect to the multifactorial etiology underlying the risk of disease of the dental hard tissue and the periodontal tissues, anterior crowding > 3 mm (threshold value) as an individual "host factor" represents a cumulative risk potential for chronic inflammatory processes whose consequences are manifest only at a higher age. This gives rise to a medical treatment indication within the framework of preventive treatment strategies. PMID- 14749888 TI - Investigations on the influencing of the subgingival microflora in chronic periodontitis. A study in adult patients during fixed appliance therapy. AB - AIM: The aim of the present study was to investigate changes in the subgingival flora in adults with chronic periodontitis undergoing orthodontic fixed appliance therapy. PATIENTS AND METHODS: In seven adult patients who had undergone nonantibiotic periodontal pretreatment, the subgingival bacteria were subjected to microbiological examination and the number of periodontopathogenic organisms was determined before (T1: prior to treatment being started), during (T2: 6 weeks after orthodontic treatment was started) and after the end of orthodontic treatment (T3: 6 weeks after removal of the fixed appliances). RESULTS AND CONCLUSION: During the fixed appliance therapy (metal brackets, NiTi archwires, stainless steel archwires), a marked reduction was observed in the total bacteria count from the subgingival pocket despite the clinical periodontal parameters remaining almost unchanged. However, the total count of some highly pathogenic bacteria rose again slightly after the end of treatment. We attribute the marked improvement in the periodontopathogenic bacteria spectrum under fixed appliance therapy with metal brackets, NiTi archwires and stainless steel archwires to metal corrosion entailing the release of primarily nickel ions, which have a toxic effect on bacteria and thus enable the regeneration of the physiological bacterial flora. In none of the patients was a deterioration of the periodontal status observed during and after fixed appliance therapy. PMID- 14749887 TI - Torque deformation characteristics of plastic brackets: a comparative study. AB - AIM: The purpose of this study was to compare the torque deformation characteristics of seven commercially available plastic brackets, both amongst each other and with stainless steel brackets. MATERIAL AND METHODS: Ten brackets each of (1) pure polycarbonate, (2) ceramic reinforced polycarbonate, (3) fiberglass reinforced polycarbonate, (4) ceramic reinforced polycarbonate with metal slot, (5) fiberglass reinforced polycarbonate with metal slot, (6) polyurethane, and (7) polyurethane with metal slot were exposed to torsion in a torquemeter, following an aging process according to ISO 10477. Ten stainless steel brackets served as a control group. Torsion was applied continuously using a material testing machine (Zwick Z2.5, Zwick Materialprufung, Ulm, Germany). RESULTS: The results showed within the group of plastic brackets that metal slot reinforced brackets were subject to the lowest degree of deformation, followed by the brackets made of pure polyurethane, pure polycarbonate and fiberglass reinforced polycarbonate. The ceramic reinforced polycarbonate brackets showed the highest deformation under torque stress. The plastic deformation of the ceramic reinforced, fiberglass reinforced and pure polycarbonate brackets started even in the recommended torque range of 10-20 Nmm. The additional ceramic and fiberglass in the plastic brackets investigated in this study did not improve the torque stability of the polycarbonate brackets. The pure polyurethane brackets showed no significant difference from the pure polycarbonate brackets at the moment of 15 Nmm which is the optimal torque for a maxillary incisor. CONCLUSION: A comparison with the stainless steel brackets illustrated that plastic brackets are suited for clinical application only if they have a metal slot. PMID- 14749889 TI - Allergies induced by orthodontic alloys: incidence and impact on treatment. Results of a survey in private orthodontic offices in the Federal State of Hesse, Germany. AB - BACKGROUND: The incidence of allergies in general is on the increase. An allergic reaction can also occur during any dental and orthodontic treatment. However, the allergic potential of orthodontic appliances is frequently overestimated. MATERIAL AND METHOD: The incidence of suspected allergic reactions during fixed appliance therapy in 68 orthodontic offices in the German State of Hesse was determined by questionnaire at approximately 0.3% of the 60,000 patients covered. RESULTS: More extraoral (45%) than intraoral (17%) skin changes were registered, with both intraoral and extraoral changes being observed in 38%. In 53% of the affected cases the therapy was adapted to nickel-free materials, whereas it was continued as planned after a brief recovery period in 33%. The treatment was discontinued in 14% of the affected patients, corresponding to one in every 3150. The individual tolerance can often be tested by inserting one bracket or one band. In addition, early orthodontic treatment seems to promote a certain immune tolerance, especially towards extraoral nickel contacts. However, if a patient is known to have a nickel allergy, materials containing nickel should be renounced on principle in the orthodontic appliances. CONCLUSION: Skin changes occurring in the course of orthodontic treatment should be examined and verified if necessary by a dermatologist. Gold plating and other coatings (titanium nitride) of the metal elements even encourage corrosion after a brief protection period. Soldering should be avoided. PMID- 14749890 TI - Orofacial development in children with Down's syndrome 12 years after early intervention with a stimulating plate. AB - BACKGROUND: Orofacial regulation therapy for children with Down's syndrome was introduced to Europe in Munich in 1978. Since then, many clinical studies have provided scientific evidence that this therapeutic approach enhances the orofacial function and facial appearance of children with trisomy 21. Only few long-term results have been published to date. PATIENTS AND RESULTS: In the present study, 20 children with trisomy 21 were examined more than 12 years after starting treatment in infancy with a Castillo Morales stimulating plate. The follow-up examination showed that the improved orofacial appearance resulting from the early treatment had remained stable in most cases. Although the mechanical stimulus of the stimulating plate was absent during the follow-up period, some patients revealed a lip and tongue posture superior to that recorded at baseline. CONCLUSION: According to the results of the present study, the orofacial status in early childhood is decisive for the subsequent development of the orofacial region and the long-term stability of the achieved improvements: Children with a pronounced orofacial dysfunction showed a greater stimulation plate-induced improvement than those with initially moderate orofacial findings. This observation was confirmed by the findings of the 12-year follow-up: Children with Down's syndrome and initially slight orofacial impairment displayed only slight improvements or unchanged findings. PMID- 14749891 TI - Use of a herbicide or lysine plus threonine for non-antibiotic selection of transgenic chickpea. AB - A desensitized aspartate kinase (AK) gene has been developed as a non-antibiotic selection marker for use in the production of transgenic chickpea ( Cicer arietinum L.). Transgenic shoots regenerated from embryo explants bombarded with the desensitized AK gene were selected on media containing two amino acids, lysine and threonine (LT). Approximately 15% of the putative transgenic shoots of vars. P-362 and P-1042 survived after 4 weeks of growth on MSB5 medium (MS mineral salts and B5 vitamins) containing 2 microM thidiazuron (TDZ) and 2 mM lysine and 2 m M threonine. These shoots were subsequently grown on MSB5 medium supplemented with 2 micro M TDZ and 5 mM lysine and 5 mM threonine, and nearly 1% continued to grow after 16 weeks of selection. A phosphinothricin (PPT) selection system for Agrobacterium-mediated chickpea transformation was also developed. Three varieties of chickpea, P-362, P-1042 and P-1043, were successfully used for Agrobacterium transformation. Following Agrobacterium infection, 3-8% of the regenerated shoots remained green and continued to grow on MSB5 medium supplemented with 2.5 mg l(-1 )PPT. Increasing the concentrations of PPT to 15 mg l(-1) reduced transgenic shoot production in P-362, P-1042 and P-1043 to 0.7%, 1.2% and 1.1%, respectively. Selected putatively transformed shoots of all three varieties were rooted and grown to maturity. Southern hybridization analysis revealed single as well as multiple integration of genes in selected transgenic lines. The level of AK activity detected in LT-selected plants was higher than that detected in the non-transformed control. PMID- 14749892 TI - Green fluorescent protein as a visual selection marker for papaya (Carica papaya L.) transformation. AB - Chemical-based selection for plant transformation is associated with a number of real and perceived problems that might be avoided through visual selection. We have used green fluorescent protein (GFP), as a visual selectable marker to produce transformed papaya ( Carica papaya) plants following microprojectile bombardment of embryogenic callus. GFP selection reduced the selection time from 3 months on a geneticin (G418) antibiotic-containing medium to 3-4 weeks. Moreover, GFP selection increased the number of transformed papaya plants by five to eightfold compared to selection in the presence of antibiotics. Overall, the use of GFP for selecting transgenic papaya lines improved our throughput for transformation by 15- to 24-fold while avoiding the drawbacks associated with the use of antibiotic resistance-based selection markers. PMID- 14749893 TI - Negative selection using thymidine kinase increases the efficiency of recovery of transformants with targeted genes in the filamentous fungus Leptosphaeria maculans. AB - A vector system was constructed that is designed to decrease the number of transformants required to be screened when looking for gene disruption events in filamentous fungi. This vector was used to mutate two genes, an ATP-binding cassette transporter ( LmABCt4) and a two-component histidine kinase gene ( LmHK1) in the ascomycete Leptosphaeria maculans. The system uses the thymidine kinase gene from the herpes simplex virus as a negative selectable marker. Thymidine kinase expression is regulated by the TrpC regulatory elements from Aspergillus nidulans and should be applicable to other ascomycetous fungi. When thymidine kinase is expressed in the presence of particular thymidine analogues, these analogues are converted to toxic compounds which kill the cell. We also report the transformation of L. maculans using Agrobacterium tumefaciens-mediated DNA delivery. PMID- 14749894 TI - Interaction pharmacokinetics of pegylated liposomal doxorubicin (Caelyx) on coadministration with paclitaxel or docetaxel. AB - PURPOSE: To investigate the pharmacokinetics of polyethylene glycol-coated liposomal doxorubicin (PLD, Caelyx) when given as a single agent and in combination with the taxanes paclitaxel or docetaxel in humans. METHODS: The plasma kinetics of PLD were studied in 19 cancer patients treated with PLD every 4 weeks combined with either paclitaxel (on a weekly basis in seven and as a single infusion in three patients) or docetaxel (weekly in seven and as a single infusion in two). Plasma concentrations of PLD were quantified in two sets of samples per patient to compare the same pharmacokinetic parameters in each subject when treated with single-agent PLD and again with the combination. Total doxorubicin concentrations in plasma were quantified by HPLC. Pharmacokinetics were evaluated by noncompartmental analysis and the data obtained were compared for differences by a matched-pairs nonparametric test. RESULTS: Coadministration of paclitaxel produced a median/mean 54/80% increase in PLD AUC(inf) (95% confidence interval 23% to 136%, P=0.002). The observed increase was consistent among all subjects. PLD clearance was also decelerated in the presence of paclitaxel (P=0.013) while other pharmacokinetic parameters were affected modestly. A small increase in the AUC of PLD was observed in the docetaxel/PLD arm (mean increase 12%, P=0.039) while PLD clearance decreased marginally and other pharmacokinetic parameters remained unaffected. AUC extrapolated to infinity was below 8% in both arms. CONCLUSIONS: This study showed the presence of a pharmacokinetic interaction that led to higher plasma concentrations of PLD when combined with paclitaxel and to a minor extent when combined with docetaxel. This pharmacokinetic information may be of value when planning combination therapies of PLD with taxanes. PMID- 14749899 TI - Assessing landowner activities related to birds across rural-to-urban landscapes. AB - Fluctuations of bird abundances in the Midwest region of the United States have been attributed to such factors as landscape change, habitat fragmentation, depredation, and supplemental feeding. However, no attempt has been made to estimate the collective role of landowner activities that may influence birds across a landscape. To investigate how landowners might influence birds when the majority (> 90%) of land is privately owned, we surveyed all 1694 private domestic landowners living on three breeding bird survey routes (approximately 120 km) that represent a continuum of rural-to-urban landscapes in Southeastern Michigan from October through December 2000. Our survey was designed to investigate (1) the proportion of landowners involved in bird feeding, providing bird houses, planting or maintaining vegetation for birds, gardening, landscaping, applying fertilizer, and applying pesticides or herbicides; (2) whether differences existed between urban, suburban, and rural landowner activities; and (3) whether landowners that carried out a given activity were sociodemographically different from those who did not. Of the 968 respondents (58.5% response rate), 912 (94%) carried out at least one of the activities on their land and the average landowner carried out 3.7 activities. A total of 65.6% fed birds, 45.7% provided bird houses, 54.6% planted or maintained vegetation for birds, 72.7% gardened, 72.3% landscaped, 49.3% applied fertilizer, and 25.2% applied pesticides or herbicides. Significant differences existed between the landscapes, with rural landowners having more bird houses and applying pesticides or herbicides in greater frequency. Similarly, urban landowners had a greater density of bird feeders and houses, but planted or maintained vegetation in the lowest frequency. Participation in activities varied by demographic factors, such as age, gender, and occupation. Scaling each activity to all landowners, including nonrespondents, across all landscapes indicates that between 14% and 82% of landowners may be engaged in a particular activity, with application of pesticides or herbicides having the least potential involvement (13.9%-55.4%) and gardening having the greatest potential involvement (40.1%-81.6%). Taken collectively, our results indicate that landowners are both intentionally and unintentionally engaged in a wide range of activities that are likely to influence bird populations. PMID- 14749900 TI - Apoptosis pathways in cancer and cancer therapy. AB - Activation of apoptosis pathways is a key mechanism by which cytotoxic drugs kill tumor cells. Also immunotherapy of tumors requires an apoptosis sensitive phenotype of target cells. Defects in apoptosis signalling contribute to resistance of tumors. Activation of apoptosis signalling following treatment with cytotoxic drugs has been shown to lead to activation of the mitochondrial (intrinsic) pathway of apoptosis. In addition, signalling through the death receptor (extrinsic) pathways, contributes to sensitivity of tumor cells towards cytotoxic treatment. Both pathways converge finally at the level of activation of caspases, the effector molecules in most forms of cell death. In addition to classical apoptosis, non-apoptotic modes of cell death have recently been identified. Mechanisms to overcome apoptosis resistance include direct targeting of antiapoptotic molecules expressed in tumors as well as re-sensitization of previously resistant tumor cells by re-expression of caspases and counteracting apoptotis inhibitory molecules such as Bcl-2 and molecules of the IAP family of endogenous caspase inhibitors. Molecular insights into regulation of apoptosis and defects in apoptosis signalling in tumor cells will provide novel approaches to define sensitivity or resistance of tumor cells towards antitumor therapy and provide new targets for rational therapeutic interventions for future therapeutic strategies. PMID- 14749902 TI - The nutrient artery canal of the femur: a radiological study in patients with primary total hip replacement. AB - OBJECTIVE: Plain antero-posterior and lateral radiographs of the femur often show radiolucent lines, which may reflect the canal of a nutrient artery. In patients who have undergone total hip replacement these radiolucent lines must be differentiated from fractures caused by bursting of the shaft during the procedure. DESIGN AND PATIENTS: In a retrospective radiological study 129 extremities of 95 patients with cementless primary hip prostheses were examined for radiolucent lines. Pre- and postoperative antero-posterior and lateral radiographs were analyzed. RESULTS: In 34 of 129 extremities (26.4%) radiolucent lines compatible with a nutrient artery canal were detected, 14 of which (10.9% overall) were seen on lateral radiographs. In 11 of 129 extremities (8.5%) a nutrient artery canal was detected only on the antero-posterior views, while in 9 of 129 extremities (7.0%) it was well defined in both projections. One patient (0.8%) presented with a fracture postoperatively, which was radiologically distinct from a nutrient artery canal. The distance between the tip of the greater trochanter and the proximal end of the nutrient artery canal was 170+/-25 mm; the canal length was 27+/-9 mm. CONCLUSION: Nutrient artery canals are often seen radiologically on pre- and postoperative radiographs down to the mid-shaft level and should be routinely recorded. PMID- 14749901 TI - Spontaneous malignant transformation of conventional giant cell tumor. AB - Spontaneous malignant transformation of conventional giant cell tumor (GCT) of bone is exceedingly rare. We report on a case of GCT of the iliac crest in a 35 year-old woman with malignant change into a high-grade osteosarcoma 10 years after the first appearance of GCT on a radiograph. Since the patient refused therapy for personal reasons the tumor remained untreated until sarcomatous transformation occurred. Image cytometry showed DNA aneuploidy and a suspiciously high 2c deviation index (2cDI) in the primary bone lesion. A thorough review of the world literature revealed only seven fully documented cases of secondary malignant GCT which matched the definition of a "sarcomatous growth that occurs at the site of a previously documented benign giant cell tumor" and not treated by radiotherapy. These cases as well as the current one suggest that a spontaneous secondary malignant GCT presents as a frankly sarcomatous tumor in the form of an osteosarcoma or malignant fibrous histiocytoma. It usually appears at sites of typical GCTs-often without any recurrent intermediate state-and is diagnosed 3 or more years after the primary bone lesion. The prognosis is poor. PMID- 14749903 TI - Principles of biorefineries. AB - Sustainable economic growth requires safe, sustainable resources for industrial production. For the future re-arrangement of a substantial economy to biological raw materials, completely new approaches in research and development, production and economy are necessary. Biorefineries combine the necessary technologies between biological raw materials and industrial intermediates and final products. The principal goal in the development of biorefineries is defined by the following: (biomass) feedstock-mix + process-mix --> product-mix. Here, particularly the combination between biotechnological and chemical conversion of substances will play an important role. Currently the "whole-crop biorefinery", "green biorefinery" and "lignocellulose-feedstock biorefinery" systems are favored in research and development. PMID- 14749904 TI - An antecedent of the MHC-linked genomic region in amphioxus. AB - The MHC genes on human chromosome 6 are located within one of the best characterised paralogy regions of the human genome. Numerous genes mapping around this location, 6p21, have paralogues at one, two or three other chromosomal locations on HSA 1, 9 and 19. The similarity between these four chromosomal regions suggests the linkages may have adaptive significance, and/or they may be echoes of segmental or genome duplication in human ancestry. Here, we show that six amphioxus cosmids, containing genes orthologous to those from the human MHC linked paralogy regions, map to a single amphioxus chromosome. The composition of the MHC-linked genomic region, therefore, pre-dates vertebrate origins. PMID- 14749905 TI - Effects of the Norwegian winter environment on Giardia cysts and Cryptosporidium oocysts. AB - The structural integrity of Cryptosporidium oocysts and Giardia cysts in the Norwegian winter environment was investigated. During winter 2001/2002, Cryptosporidium oocysts and Giardia cysts were placed in the upper layers of soil in different matrices contained in chambers and exposed to the Norwegian climate. Morphological characteristics and inclusion/exclusion of vital dyes were monitored and compared to refrigerated controls. Reduction in parasite numbers was recorded for all parasites, geographical locations, and matrices. Shear forces generated during freeze-thaw cycles are postulated to have disintegrated the parasites exposed to the Norwegian winter and retrospective laboratory studies support this theory. Increased dye inclusion, possibly indicative of viability loss, was also noted. The refrigerated control parasites exhibited no decline in numbers, and alteration in dye inclusion characteristics for refrigerated parasites was slower. Cryptosporidium oocysts were apparently more robust than Giardia cysts; differences between isolates were also noted. These results suggest Cryptosporidium oocysts and Giardia cysts do not persist in the Norwegian terrestrial environment over winter, and when detected, will have been excreted since the previous winter. Differences in the morphological characteristics, matrix effects, and the possible relationship of the dye data to parasite survival are discussed in relation to further studies. PMID- 14749906 TI - Microbial monitoring of spacecraft and associated environments. AB - Rapid microbial monitoring technologies are invaluable in assessing contamination of spacecraft and associated environments. Universal and widespread elements of microbial structure and chemistry are logical targets for assessing microbial burden. Several biomarkers such as ATP, LPS, and DNA (ribosomal or spore specific), were targeted to quantify either total bioburden or specific types of microbial contamination. The findings of these assays were compared with conventional, culture-dependent methods. This review evaluates the applicability and efficacy of some of these methods in monitoring the microbial burden of spacecraft and associated environments. Samples were collected from the surfaces of spacecraft, from surfaces of assembly facilities, and from drinking water reservoirs aboard the International Space Station (ISS). Culture-dependent techniques found species of Bacillus to be dominant on these surfaces. In contrast, rapid, culture-independent techniques revealed the presence of many Gram-positive and Gram-negative microorganisms, as well as actinomycetes and fungi. These included both cultivable and noncultivable microbes, findings further confirmed by DNA-based microbial detection techniques. Although the ISS drinking water was devoid of cultivable microbes, molecular-based techniques retrieved DNA sequences of numerous opportunistic pathogens. Each of the methods tested in this study has its advantages, and by coupling two or more of these techniques even more reliable information as to microbial burden is rapidly obtained. PMID- 14749907 TI - Microbial diversity in inactive chimney structures from deep-sea hydrothermal systems. AB - Massive chimney structures, which are characteristic of many hydrothermally active zones, harbor diverse microbial communities containing both thermophilic and hyperthermophilic microbes. However, vent chimneys ultimately become hydrothermally inactive, and the changes that occur in the microbial communities upon becoming inactive have not been documented. We thus collected inactive chimneys from two geologically and geographically distinct hydrothermal fields, Iheya North in the western Pacific Ocean and the Kairei field in the Indian Ocean. The chimneys displayed easily distinguishable strata, which were analyzed with regard to both mineralogical and microbiological properties. X-ray diffraction pattern and energy-dispersive spectroscopic analyses revealed that the main mineral components of the chimney substructures from Iheya North and the Kairei field were barite (BaSO4) and chalcopyrite (CuFeS2), respectively. Microbial cell densities in the substructures determined by DAPI counting ranged from 1.7 x 10(7) cells g(-1) to 3.0 x 10(8) cells g(-1). The proportions of archaeal rDNA in the whole microbial rDNA assemblages in all substructures were, at most, a few percent as determined by quantitative fluorogenic PCR. The microbial rDNA clone analysis and whole-cell fluorescence in situ hybridization revealed a community that was decidedly different from any communities previously reported in active chimneys. Curiously, both samples revealed the abundant presence of a group of Bacteria related to a magnetosome-bearing bacterium, " Magnetobacterium bavaricum" of the Nitrospirae division. These results suggest that inactive chimneys provide a distinct microbial habitat. PMID- 14749908 TI - Microbial characterization during the early habitation of the International Space Station. AB - An evaluation of the microbiota from air, water, and surface samples provided a baseline of microbial characterization onboard the International Space Station (ISS) to gain insight into bacterial and fungal contamination during the initial stages of construction and habitation. Using 16S genetic sequencing and rep-PCR, 63 bacterial strains were isolated for identification and fingerprinted for microbial tracking. Of the bacterial strains that were isolated and fingerprinted, 19 displayed similarity to each other. The use of these molecular tools allowed for the identification of bacteria not previously identified using automated biochemical analysis and provided a clear indication of the source of several ISS contaminants. Strains of Bradyrhizobium and Sphingomonas unable to be identified using sequencing were identified by comparison of rep-PCR DNA fingerprints. Distinct DNA fingerprints for several strains of Methylobacterium provided a clear indication of the source of an ISS water supply contaminant. Fungal and bacterial data acquired during monitoring do not suggest there is a current microbial hazard to the spacecraft, nor does any trend indicate a potential health risk. Previous spacecraft environmental analysis indicated that microbial contamination will increase with time and will require continued surveillance. PMID- 14749909 TI - Gene expression in the cyanobacterium Anabaena sp. PCC7120 under desiccation. AB - The N2-fixing cyanobacterium Anabaena sp. PCC7120 showed an inherent capacity for desiccation tolerance. A DNA microarray covering almost the entire genome of Anabaena was used to determine the genome-wide gene expression under desiccation. RNA was extracted from cells at intervals starting from early to late desiccation. The pattern of gene expression in DNA fragments was categorized into seven types, which include four types of up-regulated and three types of down regulated fragments. Validation of the data was carried out by RT-PCR on selected up-regulated DNA fragments and was consistent with the changes in mRNA levels. Our conclusions regarding desiccation tolerance for Anabaena sp. PCC7120 are as follows: (i) Genes for osmoprotectant metabolisms and the K+ transporting system are up-regulated from early to mid-desiccation; (ii) genes induced by osmotic, salt, and low-temperature stress are up-regulated under desiccation; (iii) genes for heat shock proteins are up-regulated after mid-desiccation; (iv) genes for photosynthesis and the nitrogen-transporting system are down-regulated during early desiccation; and (v) genes for RNA polymerase and ribosomal protein are down-regulated between the early and the middle phase of desiccation. Profiles of gene expression are discussed in relation to desiccation acclimation. PMID- 14749910 TI - Outcomes after "unroofing" of a myocardial bridge of the left anterior descending coronary artery in children with hypertrophic cardiomyopathy. AB - The importance of myocardial bridging of a coronary artery in the setting of hypertrophic cardiomyopathy is controversial, although we have previously reported an association with myocardial ischemia and sudden death in children. We report five symptomatic children in whom symptoms or evidence of myocardial ischemia resolved or improved after supraarterial myotomy alone. PMID- 14749911 TI - Diffusion-weighted MRI in cortical ischaemia. AB - We carried out MRI on 16 male and three female comatose patients, aged 2 days to 79 years, with suspected cortical ischaemia referred from our intensive care units. Using a head coil, and following standard imaging, including coronal fluid attenuated inversion-recovery images, we performed diffusion-weighted imaging (DWI) using a whole-brain multislice single-shot echo-planar sequence with b 0 and 1000 s/mm2: 5-mm slices covering the whole brain, TR 7000 TE 106 ms, 128 x 128 pixels, field of view 250 mm, one excitation. Maps of apparent diffusion coefficients (ADC) were generated automatically. DWI showed cortical, basal ganglia and watershed-area high signal in all cases, associated with a decrease in ADC to 60- 80% of normal. DWI showed lesions not seen (40%) or underestimated (40%) on conventional T2-weighted imaging. Within 24 h of the onset of symptoms, DWI showed changes not readily detectable on T2-weighted images. The cortical high signal on DWI and the ADC changes, suggesting severe ischaemia rather than oedema, was found in areas known to be affected by cortical laminar necrosis. Extension to the brain stem and white matter was associated with a higher likelihood of death. PMID- 14749912 TI - Pharmacological treatment of acute otitis media in children. A comparison among seven locations: Tenerife, Barcelona and Valencia (Spain), Toulouse (France), Smolensk (Russia), Bratislava (Slovakia) and Sofia (Bulgaria). AB - OBJECTIVE: To describe patterns observed in the treatment of acute otitis media (AOM) in several locations of five countries. PATIENTS AND METHODS: Cross sectional, descriptive study. Random sample of 12,264 paediatric outpatients seen by paediatricians or general practitioners (GPs). Data on patient demographics, diagnoses and treatment were collected. Diagnoses were coded by ICD-9 and drugs by ATC classification. Patients diagnosed with AOM (ICD-9 codes: 381 and 382) were selected for analysis. RESULTS: Cases of AOM (873) accounted for 7.1% of the sample. There is a clear variation in the percentage of children diagnosed with AOM and treated with antibiotics in the different locations, antibiotic prescriptions being higher in Barcelona (93% of children), and lowest in Smolensk (56.4 % of children were treated without antibiotics). The antibiotics used varied widely: ampicillin use is almost limited to Smolensk (26.7%) and Bratislava (13.8%), whereas amoxicillin plus clavulanic acid is the choice in Toulouse (33.8%), Valencia (30.2%) and Barcelona (28.9%), and cephalosporins are more frequently prescribed in Tenerife (51.7%). Finally, macrolides are used in Barcelona (18.3%), Valencia (17.5%) and Tenerife (13.6%), but not prescribed in Toulouse or Sofia. Prescriptions of anti-inflammatory drugs were only relevant in Valencia (31.7%), Tenerife (27.2%) and Toulouse (17.4%) and of otological preparations in Sofia, where almost each child received ear drops (91.9%). Nasal preparations are commonly used only in Sofia (41.9%), Bratislava (65.5%) and Smolensk (68.6%). CONCLUSION: Despite the general agreement of most guidelines, wide differences in the treatment of uncomplicated AOM in children are observed. Non-antibiotic therapy for AOM and the use of first-choice antibiotics should be more actively encouraged in the primary care centres. Studies to measure prevailing rates of antibiotic resistance in these populations are needed. PMID- 14749913 TI - Polybrominated diphenyl ether congeners and toxaphene in selected marine standard reference materials. AB - Polybrominated diphenyl ether (PBDE) congeners and components of the complex mixture toxaphene are stable in the environment and readily bioaccumulated into wildlife and human tissues. PBDEs are presently used in large quantities worldwide as flame retardants in textiles, furniture, computer equipment, and cables. Toxaphene is a complex mixture of chlorinated bornanes and bornenes that was the most heavily used pesticide in the United States until it was banned in 1982; however, some countries continue to use toxaphene. The National Institute of Standards and Technology has quantified PBDE congeners and toxaphene in several available Standard Reference Materials (SRMs) using methods of gas chromatography with electron impact mass spectrometry (GC-EI-MS) and GC negative chemical ionization (NCI) MS, respectively. SRM 1588a Organics in Cod Liver Oil and SRM 1945 Organics in Whale Blubber were examined for PBDE congeners 47, 99, 100, 153, and 154, total toxaphene, and toxaphene congeners 26, 50, and 62. SRM 1946 Lake Superior Fish Tissue was also examined for total toxaphene and toxaphene congeners. The sum of the PBDE congeners (mean, (1 SD) wet basis) for SRM 1945 was 150 ng g(-1) (7 ng g(-1)). The concentration of PBDE 47 in SRM 1588a was 82.7 ng g(-1) (2.8 ng g(-1)). Other PBDEs were detected in SRM 1588a but were not quantified due their low levels. The total toxaphene (wet mass basis) was 1,210 ng g(-1) (127 ng g(-1)), 1,960 ng g(-1) (133 ng g(-1)), and 3,980 ng g(-1) (248 ng g(-1)) in SRMs 1945, 1946, and 1588a, respectively. The values for PBDEs and toxaphene determined in the SRMs, while not certified, indicate that the SRMs will be suitable control materials for PBDE and toxaphene analyses. PMID- 14749914 TI - Correlation between biomarkers of polycyclic aromatic hydrocarbon exposure and electrophilic tissue burden in a rat model. AB - This study was aimed at investigating the correlation between biomarkers of exposure to polycyclic aromatic hydrocarbons and, more specifically, at examining the role of urinary 1-hydroxypyrene (1-OHP) as a reliable measure of internal dose linked to the electrophilic tissue burden (ETB), assessed as covalent binding of the ultimate carcinogen benzo( a)pyrene diolepoxide (BaPDE) with cellular proteins in target organs. The protocol included experimental verification of a previously proposed algorithm for adjustment of reference values for urinary 1-OHP with exposure to different mixtures of polycyclic aromatic hydrocarbons in a rat model. Hence, the relationships between ETB in liver, lung, and heart as well as the BaPDE-haemoglobin adducts level on the one hand, and urinary/faecal 1-OHP or urinary/faecal 3-hydroxybenzo( a)pyrene (3 OHBaP) on the other hand have been examined. Male Sprague-Dawley rats received intraperitoneally, once daily for 10 consecutive days, binary mixtures of benzo( a)pyrene (BaP) and pyrene (P) in three different exposure scenarios corresponding to BaP/P ratios of 0.2, 1 and 5, with three doses of BaP (2, 6 and 20 mg/kg) for each scenario. The ETB levels were measured as the ultimate analyte benzo( a)pyrene tetrol (BaPTeT) obtained after mild acid hydrolysis of BaPDE adducts with proteins. It was experimentally confirmed that: (1) urinary 1-OHP is a reliable biomarker linked to the ETB in tissues that are targets for carcinogenicity, such as lung, for the BaP/P ratios of 0.2 and 1 (linear regression p=0.0099 and 0.0293, respectively); (2) urinary 3-OHBaP is correlated with the BaPDE-haemoglobin adducts for all three exposure scenarios ( p=0.0011 for BaP/P=0.2, p<0.0001 for BaP/P=1 and p=0.0099 for BaP/P=5). The experimental relationship between ETB and urinary 1-OHP was used to interpolate biological limit values for the urinary metabolite assuming three arbitrary critical levels of ETB. These were compared with the values calculated from the algorithm using the BaP/P ratio 1 mixture as a reference. The ratios of calculated to observed values varied from 1.0 to 1.6 for the BaP/P 0.2 mixture, and from 1.9 to 3.0 for the BaP/P 5 mixture. The results obtained in the present study indicate that the algorithm mentioned above applies well for two of the three exposure scenarios corresponding to realistic occupational BaP/P ratios of 0.2 and 1. This suggests that, using ETB as an endpoint, the proposed algorithm will reasonably predict the critical value of urinary 1-OHP for mixtures having different BaP/P ratios. Stronger linear relationships between ETB in all chosen tissues and 1-OHP or 3 OHBaP excretion were obtained with urinary metabolites than with their faecal analogues. Thus urinary 1-OHP and 3-OHBaP are more reliable biomarkers in biological monitoring strategies. PMID- 14749915 TI - Mutational analysis of feedback inhibition and catalytic sites of prephenate dehydratase from Corynebacterium glutamicum. AB - Prephenate dehydratase is a key regulatory enzyme in the phenylalanine-specific pathway of Corynebacterium glutamicum. PCR-based random mutagenesis and functional complementation were used to screen for m-fluorophenylalanine ( mFP) resistant mutants. Comparison of the amino acid sequence of the mutant prephenate dehydratases indicated that Ser-99 plays a role in the feedback regulation of the enzyme. When Ser-99 of the wild-type enzyme was replaced by Met, the specific activity of the mutant enzyme was 30% lower than that of the wild-type. The Ser99Met mutant was active in the presence of 50 microM phenylalanine, whereas the wild-type enzyme was not. The functional roles of the eight conserved residues of prephenate dehydratase were investigated by site-directed mutagenesis. Glu64Asp substitution reduced enzyme activity by 15%, with a 4.5- and 1.7-fold increase in Km and kcat values, respectively. Replacement of Thr-183 by either Ala or Tyr resulted in a complete loss of enzyme activity. Substitution of Arg-184 with Leu resulted in a 50% decrease of enzyme activity. The specific activity for Phe185Tyr was more than 96% lower than that of the wild-type, and the Km value was 26-fold higher. Alterations in the conserved Asp-76, Glu-89, His 115, and Arg-236 residues did not cause a significant change in the Km and kcat values. These results indicated that Glu-64, Thr-183, Arg-184, and Phe-185 residues might be involved in substrate binding and/or catalytic activity. PMID- 14749916 TI - Localized pigmented villonodular synovitis in the anteromedial compartment of the knee associated with cartilage lesions of the medial femoral condyle: report of a case and review of the literature. AB - Localized pigmented villonodular synovitis (PVNS) of the knee is an uncommon entity, presenting with different clinical signs and symptoms. We report on a case of a 42-year-old woman who presented with a 3-year history of knee pain and mechanical problems such as locking. On examination she was found to have a palpable and painful mass over the anteromedial joint line. Magnetic resonance imaging (MRI) revealed a soft tissue mass in the anteromedial compartment of the knee joint. The lesion was completely resected arthroscopically, and histologic examination confirmed the diagnosis of localized PVNS. The patient was free of symptoms, and MRI examination showed no evidence of recurrence at 1-year follow up. PMID- 14749917 TI - Dislocation of the knee. PMID- 14749918 TI - Serum lipids in children 3 to 5 years after kidney, liver, and heart transplantation. AB - Although dyslipidemia is common after solid organ transplantation (Tx), there are few long-term studies in children. We investigated the prevalence of dyslipidemia up to 5 years after Tx in 125 children on triple immunosuppression with one of three different well-functioning grafts, kidney, liver, and heart, and 181 controls. Total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), and triglyceride (TG) concentrations were measured annually. Low-density lipoprotein cholesterol concentrations were also calculated. The risk factors for dyslipidemia were determined at 3 years. There was a high prevalence of hypertriglyceridaemia in all three groups, 50% in the kidney transplantation (KTx) and heart transplantation (HTx) groups and 30% in the liver transplantation (LTx) group. In addition, 50% of KTx patients had high TC. In the Tx groups taken together, the following independent associations were observed: KTx and high pre Tx TC were associated with high TC, high trough concentration of blood cyclosporine with low HDL-C, and older age at Tx accounted for higher TG. Dyslipidemia, especially hypertriglyceridaemia, was common 3-5 years after Tx. The aetiology is multifactorial and depends on the transplanted organ. PMID- 14749919 TI - Abbreviated mycophenolic acid AUC from C0, C1, C2, and C4 is preferable in children after renal transplantation on mycophenolate mofetil and tacrolimus therapy. AB - In order to allow a similar algorithm to be used for both adults and children on tacrolimus-based and mycophenolate mofetil [MMF, a pro-drug for mycophenolic acid (MPA)]-based immunosuppression, a limited sampling technique from the trough level (C0) and the levels 30 min (C0.5) and 2 h (C2) after intake was to be developed from MPA area under the time-concentration curves (AUC). We retrospectively analyzed 49 full ten-point pharmacokinetic (PK) profiles from 29 pediatric patients on MMF and tacrolimus. We used stepwise multiple regression analysis to calculate limited sampling approaches. Agreement with the AUC was tested by means of Bland and Altman analysis. The correlation between AUC and pre dose trough concentration was r(2)=0.5188 ( P<0.0001) and between AUC and post dose trough concentration r(2)=0.6924 ( P<0.0001). The next best correlations were with 2 h (C2, r(2)=0.6711, P<0.0001), 4 h (C4, r(2)=0.6411, P<0.0001), 1.5 h (C1.5, r(2)=0.6344, P<0.0001), and 6 h (C6, r(2)=0.6219, P<0.0001). Three-point estimates at C0, C0.5, and C2 resulted in an acceptable correlation between predicted AUC and AUC from the full profile when we used the formula AUC = 10.01391+3.94791xC0+3.24253xC0.5+1.0108xC2, Pearson's r=0.8996, 95% confidence interval 0.8277-0.9424. However, even better results could be obtained when we used AUC = 8.217+3.163xC0+0.994xC1+1.334xC2+4.183xC4, Pearson's r=0.9456, 95% confidence interval 0.9051-0.9691. Bland and Altman analysis revealed good agreement between AUC predicted from C0, C0.5, and C2 and AUC from the full profile, but was inferior to the four-point approach. Also, the previously reported formula derived for adults was not usable in these patients. A special formula must be used for children. The AUC of MPA can be predicted by limited sampling including C0, C0.5, and C2, while an approach using C0, C1, C2, and C4 is preferable. PMID- 14749921 TI - Novel SBDS mutations caused by gene conversion in Japanese patients with Shwachman-Diamond syndrome. AB - Shwachman-Diamond syndrome (SDS; OMIM 260400) is an autosomal recessive disorder characterized by exocrine pancreatic insufficiency, bone marrow dysfunction and metaphyseal chondrodysplasia. SDS is caused by mutations in SBDS, an uncharacterized gene. A previous study in SDS patients largely of European ancestry found that most SBDS mutations occurred within a approximately 240-bp region of exon 2 and resulted from gene conversion due to recombination with a pseudogene, SBDSP. It is unknown, however, whether these findings are applicable to other ethnic groups. To address this question, we examined SBDS mutations in six Japanese families with SDS by direct sequencing. We identified compound heterozygous mutations in four families: two were recurrent (96-97insA, 258+2T>C), and three were novel [292-295delAAAG, (183-184TA>CT +201A>G), (141C>T+183-184TA>CT+201A>G)] mutations. Most of these mutations also appear to result from gene conversion, but the conversion events occurred at various sites between intron 1 and exon 3. Thus, gene conversion mutations in SBDS are common to different ethnic groups, but they are not confined to a limited region of the gene. PMID- 14749922 TI - Leukotriene-related gene polymorphisms in ASA-intolerant asthma: an association with a haplotype of 5-lipoxygenase. AB - A recent study has demonstrated the possible involvement of a leukotriene C4 synthase (LTC4S) gene polymorphism in ASA-intolerant asthma (AIA) in a Polish population, whereas no significant association was noted in other populations. To investigate the role of genetic polymorphism in AIA development, we screened single nucleotide polymorphisms (SNPs) of the key enzymes involved in arachidonate metabolism, and the cysteinyl leukotriene receptor 1 (CYSLTR1) in a large Korean population with AIA: 93 AIA and 181 ASA-tolerant asthma (ATA) patients, and 123 normal controls. The single-base extension method was used to genotype SNPs in 5-lipoxygenase (ALOX5, -1708G-->A, 21C-->T, 270G-->A, 1728G- >A), ALOX5-activating protein (ALOX5AP, 218A-->G), prostaglandin-endoperoxide synthase 2 (PTGS2, COX2, -162C-->G, 10T-->G, R228H, V511A), LTC4S (-444A-->C), and CYSLTR1 (927T-->C). Haplotype analyses were undertaken for the SNPs in ALOX5. No significant differences in allele and genotype frequencies of single SNPs were observed between the patient groups ( P>0.05). However, the frequency of the ALOX5-ht1[G-C-G-A] haplotype in the AIA group was significantly higher than its frequency in the ATA group with a probability ( P) of 0.01, odds ratio (OR) of 5.0, and 95% confidence interval (95%CI) of 1.54-17.9, and in the normal controls ( P=0.03, OR=4.5, 95%CI=1.1-18.4), by using a dominant model. These results suggest a lack of association between the ALOX5AP, PTGS2, LTC4S, and CYSLTR1 gene polymorphisms and the AIA phenotype in the Korean population. However, the possible involvement of ALOX5-ht1[G-C-G-A] in AIA development is suggested. PMID- 14749924 TI - Wheeze and urban variation in South Asia. AB - Typically, urban South Asia is characterised by narrow streets, heavy traffic, visible haze, unplanned city architecture and the use of kerosene or wood-burning stoves at home. However, some urban areas in South Asia are clean and modern, with concrete housing, vehicle emission regulations and use of smoke-free household fuel. The purpose of this study was to compare wheeze prevalence in 13 14-year-olds between two South Asian cities (Galle, Chandigarh), representing each of the above two archetypes. The validated one-page International Study on Asthma and Allergies in Childhood (ISAAC) questionnaire for 13-14-year-olds was used for the study. Of 1814 distributed questionnaires, 1737 (95.8%) were completed correctly and returned (Galle: n=1162; Chandigarh: n=575). Crude prevalence rates, and prevalence odds ratios (with 95% two-sided confidence intervals (CI) for comparison of prevalence rates), were calculated. The prevalence rate for wheezing in Galle (28.7%) was higher than in Chandigarh (12.5%). The prevalence odds ratios (Galle versus Chandigarh) for lifetime wheezing (2.3; 1.8-2.9), wheezing in the previous year (2.1; 1.6-2.7), exercise related wheeze (4.8; 3.5-6.7) and physician-diagnosed wheeze (1.7; 1.2-2.3) demonstrated significant differences in wheeze prevalence between the two cities ( P<0.05). The numbers of 13-14-year-olds experiencing less than 12 wheezing episodes per year or sleep disturbance due to wheeze of less than 1 night per week were also significantly higher for Galle than Chandigarh. There is a higher prevalence of wheeze in 13-14-year-old children living in an old-fashioned, congested city than in a clean and modern city in South Asia. PMID- 14749925 TI - A study of bone turnover markers in prepubertal children with phenylketonuria. PMID- 14749923 TI - Detection of telomerase activity in prostate massage samples improves differentiating prostate cancer from benign prostatic hyperplasia. AB - PURPOSE: We performed a case-control study in which we tested the ability of a non-invasive assay to detect telomerase activity and to distinguish between prostatic cancer (Pca) and benign prostatic hyperplasia (BPH) on samples of epithelial cells obtained after prostatic massage. METHODS: Telomerase activity was determined by a telomeric repeat amplification protocol (TRAP) assay. We selected 60 patients with histologically proven Pca (30 cases) or BPH (30 cases). Specimens included in this study were from patients who had no suspicious findings on digital rectal examination for cancer, had clinical evidence of lower urinary tract symptoms, had no sonographic signs of Pca at the transrectal ultrasound evaluation, had total PSA values moderately elevated (2.6-15 ng/ml), and had no evidence of other urological cancers. The whole procedure was conducted in double blind between pathologists and molecular biology operators. RESULTS: Telomerase activity was detected in 90% of Pca cases and in 13% of BPH cases. The sensitivity (90%) and specificity (76%) of this method were calculated. The positive predictive value, negative predictive value, and diagnostic efficiency were 87%, 90%, and 88% respectively. CONCLUSION: Our data indicate that telomerase activity detected by TRAP assay on prostate epithelial cells collected by prostate massage can substantially improve the distinction between Pca and BPH conditions. One of the clinical benefits resulting from the use of this new assay would be to refine the biopsy indication and to avoid for several patients without Pca the unnecessary cost and the complications of prostate biopsy. PMID- 14749926 TI - A 1-year old girl with fever and a unilateral swelling of the jaw. PMID- 14749927 TI - Quantitative morphometric analysis of the submucous plexus in age-related control groups. AB - An increased number and density of the so-called "giant ganglia" (seven or greater ganglion cells per ganglion) serve as histopathological criteria for a bowel motility disorder called intestinal neuronal dysplasia of the submucous plexus (IND B). However, because these morphological criteria have been defined based upon observations in constipated patients, the diagnostic value of previous studies is open to controversy. Moreover, no age-related reference data from unaffected controls are available. This study reports on data from unaffected controls on the variability of size and distribution of ganglia in the submucous plexus during development. Therefore, for the first time, the normal status has been defined. Four age groups have been defined: (a) premature births, gestational age less than 35 weeks; (b) 1-365 days; (c) 1-14 years and (d) 15 years to greater than 70 years). All of these groups revealed giant ganglia in the submucous plexus. With advancing age, there was a decrease in the number of giant ganglia (from 32.7% in group a to 11.2% in group d) accompanied by an inverse increase in the mean distance between all ganglia (from 0.52 mm in group a to 1.17 mm in group d). The data presented permit the conclusion that the criteria mentioned above are not apt to define IND B as an entity, since they do not allow a sufficient demarcation from the age-correlated normal values presented here. PMID- 14749929 TI - Presidential welcome address. PMID- 14749928 TI - Immunohistochemical characterization of fibroblast subpopulations in normal peritoneal tissue and in peritoneal dialysis-induced fibrosis. AB - Peritoneal fibrosis is one of the most common morphological changes observed in continuous ambulatory peritoneal dialysis (CAPD) patients. Both resident fibroblasts and new fibroblast-like cells derived from the mesothelium by epithelial-to-mesenchymal transition are the main cells involved fibrogenesis. In order to establish markers of peritoneal impairment and pathogenic clues to explain the fibrogenic process, we conducted an immunohistochemical study focused on peritoneal fibroblasts. Parietal peritoneal biopsies were collected from four patient groups: normal controls ( n = 15), non-CAPD uremic patients ( n = 17), uremic patients on CAPD ( n = 27) and non-renal patients with inguinal hernia ( n = 12). To study myofibroblastic conversion of mesothelial cells, alpha-smooth muscle actin (SMA), desmin, cytokeratins and E-cadherin were analyzed. The expression of CD34 by fibroblasts was also analyzed. Fibroblasts from controls and non-CAPD uremic patients showed expression of CD34, but no myofibroblastic or mesothelial markers. The opposite pattern was present during CAPD-related fibrosis. Expression of cytokeratins and E-cadherin by fibroblast-like cells and alpha-SMA by mesothelial and stromal cells supports that mesothelial-to myofibroblast transition occurs during CAPD. Loss of CD34 expression correlated with the degree of peritoneal fibrosis. The immunophenotype of fibroblasts varies during the progression of fibrosis. Myofibroblasts seem to derive from both activation of resident fibroblasts and local conversion of mesothelial cells. PMID- 14749930 TI - Cognitive performance and subjective experience during combined exposures to whole-body vibration and noise. AB - OBJECTIVE: The goal of this study was to examine the effects of noise and whole body vibration, individually and combined, and at various stimulus intensity levels, on cognitive performance and subjective experience. METHOD: Fifty-four participants (27 men and 27 women) with a mean age of 25 years, ranging from 19 to 30, were exposed for 20 min each to a 16-Hz sinusoidal whole-body vibration, a helicopter sound at 21 Hz, both stimuli combined, and a control condition. Participants were randomly assigned to one of three groups: low intensity [77 dB(A) noise and 1.0 m/s2 vibration], medium intensity [81 dB(A)/1.6 m/s2] or high intensity [86 dB(A)/2.5 m/s2. During each environmental exposure, short-term memory performance was tested with a visual Sternberg paradigm. Reaction time was measured as a dependent variable. Directly following each environmental exposure, participants rated the difficulty of the task and the annoyance level of the exposure stimulus. RESULTS: Results revealed no significant changes in reaction times due to environmental exposure or intensity level. However, participants significantly rated the combined exposure as both more annoying and more difficult than the other conditions. Further, the high-intensity group rated subjective annoyance significantly higher than the other groups for all conditions. CONCLUSIONS: The results from this study indicate that performance alone is not a sufficient measure for the study of the effects of combined stimuli on a human operator. PMID- 14749931 TI - Distribution of extracellular matrix proteins in pterygia: an immunohistochemical study. AB - PURPOSE: This study was carried out to monitor the expression of extracellular matrix proteins (ECMs) and metalloproteinases (MMPs) in pterygial tissue. METHODS: Twenty primary nasal pterygia were studied by indirect routine immunohistochemistry using 13 different primary antibodies against 8 ECMs (five collagens, fibronectin, heparan sulfate, and laminin) fibroblast growth factor (bFGF), von Willebrand factor (vWF), and 3 MMPs (8, 9, and 13). Secondary antibodies were fluoresceinated. Intensity of reaction on individual sections was graded semi-quantitatively. RESULTS: No expression of collagens I, II, and VII was found. Antibodies against collagen III reacted strongly positively (+++) with the entire pterygial stroma. Collagen IV expression was strongly positive in the wall of pterygial blood vessels, moderately positive (++) in the epithelial basement membrane, and only weakly positive (+) all over the stroma. Antibodies against fibronectin reacted moderately positively with stroma, blood vessel walls and epithelial basement membrane. Heparan sulfate was strongly expressed in the blood vessel walls and epithelial basement membrane. Antibodies against bFGF reacted only with pterygial epithelium. Laminin was strongly expressed in blood vessel wall, moderately (++) in the epithelial basement membrane and weakly over the entire stroma. vWF was strongly positive (+++) with pterygial blood vessel walls. Antibody reactions for MMPs differed. It was strong with pterygial epithelium (MMPs 8, 9 and 13), strong to moderate with pterygial stroma (MMPs 8 and 13 versus 9), and absent to weak with pterygial vascular walls (MMPs 8 and 13 versus 9). CONCLUSIONS: This study documents the presence of several ECMs but excludes the expression of others in pterygial tissues. The results especially indicate an active involvement of MMPs 8, 9 and 13 in the pathogenesis of pterygia. PMID- 14749933 TI - Infracyanine-assisted internal limiting membrane peeling in macular hole repair: does it make a difference? AB - PURPOSE: To investigate whether Infracyanine-assisted internal limiting membrane (ILM) peeling in macular hole repair affects the anatomical and functional outcome and complication rate. METHODS: A retrospective analysis of 139 consecutive cases of stage 2 and stage 3 macular hole repairs with or without Infracyanine-assisted ILM peeling. RESULTS: There were 67 eyes with Infracyanine staining of the ILM and 72 eyes in the non-ILM-stained group. The results are analyzed functionally, anatomically and in terms of complications. A similar anatomical and functional closure rate was obtained with one unclosed macular hole in the stained group and two in the non-stained group (P=1.0). Fifty-two percent of the eyes in the stained group and 59% of those in the non-stained group achieved 0.5 vision or better (P=0.40). The mean postoperative vision was 0.55 in the stained group and 0.54 in the non-stained group (P=0.65). The mean improvement in vision was 0.40 in the stained group and 0.36 in the non-stained group (P=0.21). CONCLUSION: Anatomical and functional results are similar with or without Infracyanine-assisted ILM peeling. There was no apparent adverse effect of Infracyanine use on visual function, and no apparent retinal toxicity was found. PMID- 14749932 TI - The expression of myocilin during murine eye development. AB - PURPOSE: To study the expression and localization of myocilin in the developing mouse eye. Myocilin is a 55- to 57-kDa secreted glycoprotein that is mutated in some forms of primary open-angle glaucoma. METHODS: The eyes of NMRI mice were studied from embryonic day (E) 14.5 to postnatal day (P) 21, and at 2-3 months of age. Immunohistochemistry was performed with antibodies against myocilin. The specificity of the antibodies was checked by two-dimensional gel electrophoresis. RNA was isolated from eyes at various ages, and the presence of myocilin mRNA was analyzed by northern blot hybridization. RESULTS: No immunostaining for myocilin was seen before E16.5. At around E17.5, a distinct positive immunoreactivity of optic nerve axons in the developing nerve fiber layer of the retina was observed. At P5-6, immunostaining appeared in perikarya of optic nerve ganglion cells. In the anterior eye, no immunoreactivity was observed until P10. At P12-14, the cells of the epithelial layers of ciliary body and iris, as well as the cells of the trabecular meshwork and iris stroma, became immunoreactive for myocilin. At that time, positive staining for myocilin was also seen in the corneal endothelium and in keratocytes of the corneal stroma. An essentially similar staining pattern was seen in adult eyes. Northern blot analysis for myocilin mRNA in RNA from developing mouse eyes was negative until P9. At P12, a distinct band was observed. A band with similar mobility, but somewhat more intense, was detected in mRNA from adult mouse eyes 2-3 months of age. CONCLUSIONS: The onset of immunoreactivity for myocilin in the retina occurs in parallel with the maturation of optic nerve ganglion cells. In the anterior eye, the expression of myocilin is associated with the final development of those tissues that are directly involved in aqueous humor dynamics. The presence of myocilin might be important for proper function and structure of mature optic nerve ganglion cells and aqueous humor outflow. PMID- 14749934 TI - Colonization of an intralobar pulmonary sequestration by Aspergillus. AB - Aspergillus is an opportunistic fungus that usually colonizes preexisting lung cavities, especially tuberculous ones. Colonization of a pulmonary sequestration by this germ is exceptional, with just 14 cases reported in the world literature, most of them in Asia. A case is presented of a 48-year-old woman with pleuritic thoracic pain. Simple chest radiology revealed a lower right pulmonary tumor with clear margins and a calcium-type density. CT showed it to correspond to a 6 x 5 cm hypodense mass, which was enhanced at the periphery with intravenous contrast. Aspiration puncture yielded a greenish-yellow pus and the microscopic study strongly suggested Aspergillus, confirmed by culture as Aspergillus fumigatus. Surgery revealed an infected pulmonary sequestration at the lower right lobe, and a lobectomy was performed. PMID- 14749935 TI - Follicular bronchiolitis: clinical and pathologic findings in six patients. AB - The purpose of this study was to review our experience with patients who had a definitive diagnosis of follicular bronchiolitis (FB), and to describe in detail the clinical and pathological findings, looking for common clinical aspects that may help to identify this entity. Ours is a community 750 bed teaching hospital that acts as a tertiary referral center for several subspecialties, including thoracic surgery. Six patients with a morphological diagnosis of FB, defined by the presence of coalescent germinal centers adjacent to airways, were included. Lung biopsy was obtained by thoracotomy in all patients (2 women and 4 men, mean age 53 years). In one patient FB was associated with advanced AIDS, and in another with prolonged exposure to polyethylene-flock. In 4 patients no condition previously associated with FB was found. Five patients had a history of repeated respiratory infections, 3 patients complained of dyspnea and none had peripheral blood eosinophilia. After a mean follow-up of 25 months, 2 patients responded well to steroid therapy; 3 patients suffered symptomatic exacerbations that required an increase in the steroid dose and 1 patient was not treated with steroids. The most important contribution of this series is the description of a subset of patients with FB who were not associated with other processes. These patients present relatively homogeneous clinical and pathological pictures that do not differ greatly from secondary forms. PMID- 14749936 TI - Ventricular arrhythmias and autonomic profile in patients with primary pulmonary hypertension. AB - The aim of our study was to assess the arrhythmic profile in patients with primary pulmonary hypertension (PPH) and its correlation with autonomic features, echocardiographic indexes and pulmonary function. We studied 9 subjects with a mean age of 42 +/- 11 years. All underwent echocardiography, 24-hour Holter monitoring, and cardiopulmonary exercise testing. Left ventricle ejection fraction was normal (65 +/- 6%). The right ventricle end diastolic volume was increased (108 +/- 32 ml/m2) with a slight reduction of ejection fraction (49 +/- 5%). Right ventricle systolic pressure was increased (91 +/- 25 mmHg). Heart rate variability analysis showed evidence of a reduced standard deviation of all NN intervals (SDNN) compared with the control group (102.8 +/- 32 versus 156.1 +/- 32, p < 0.005). Patients with significant ventricular arrhythmias had a lower SDNN, and lower baseline and effort PO2 (SDNN: 87.0 +/- 15 versus 115.4 +/- 38; baseline PO2: 63.2 +/- 12% versus 78.8 +/- 7%; effort PO2: 50.7 +/- 13% versus 68.7 +/- 19%). The patients with SDNN lower than 90 ms were characterized by a higher right ventricle systolic pressure (115.0 +/- 22.9 mmHg versus 79.2 +/- 17.8 mmHg, p = 0.035). The patients who experienced syncope had higher SDNN (131.7 +/- 36 versus 88.4 +/- 20, p < 0.05), higher effort PO2 (77.5 +/- 14 mmHg versus 52.3 +/- 14 mmHg, p < 0.03). The patients with PPH evidenced an increased sympathetic activity. Premature ventricular beats were more frequent in those subjects with higher adrenergic drive and lower oxygen saturation. Patients with episodes of syncope seem to have a relatively higher vagal activity, and effective mechanisms of adjustment in blood oxygenation during effort. PMID- 14749937 TI - Endostatin levels in exudative pleural effusions. AB - Endostatin is an angiogenesis inhibitor that is an endogenously produced proteolytic fragment of type XVIII collagen. Although serum levels of endostatin have extensively been studied in patients with malignant diseases, endostatin in pleural effusion has not been fully evaluated. In order to determine whether endostatin is present in pleural effusion, and to determine whether endostatin levels vary in pleural effusion of different etiology, we measured levels of endostatin in 38 malignant pleural effusion due to lung cancer patients and 29 patients with non-malignant disease using an ELISA kit. Free form of endostatin was measurable (> 11.2 pg/ml) in 26 of 38 malignant and 13 of 29 non-malignant pleural effusion. Endostatin levels in the 38 malignant pleural effusion were significantly higher than those in patients with the 29 patients with non malignant diseases ( p = 0.0131). However, there was not statistically significant difference between the patients with pleuropneumonia and those with tuberculous pleurisy ( p = 0.2194). In malignant pleural effusion due to lung cancer, the pleural effusion endostatin levels did not differ when the histological types of lung cancer were considered ( p = 0.0674). Endostatin was present in both malignant and non-malignant pleural effusion, and elevated levels of endostatin were observed in malignant pleural effusion. Although the mechanisms are unclear, elevated levels of endostatin in pleural effusion may represent the local productions of endostatin in pleural space. PMID- 14749938 TI - Effects of hyperoxia on tumor necrosis factor alpha and Grobeta expression in newborn rabbit lungs. AB - Chemokines have been implicated in the pathogenesis of many inflammatory processes, including bronchopulmonary dysplasia in mechanically ventilated premature infants. We hypothesized that early expression of the proinflammatory cytokine, tumor necrosis factor alpha (TNFalpha), would be followed by later expression of the downstream chemokine, Grobeta, in the oxygen-injured newborn lung. Reverse transcriptase-polymerase chain reaction (RT-PCR) and ribonuclease protection assay (RPA) were used to assess TNFalpha and Grobeta mRNA expression in lung RNA samples from newborn rabbits exposed to > 95% O2 for 8-9 days, followed by 60% O2 for a further 2-4 weeks or from control rabbits exposed to air. Four lung samples per condition were collected every 2 days from day 0 to day 14, and at days 22 and 36. Rabbit alveolar macrophages (AM) stimulated in vitro with bacterial lipopolysaccharide served as positive controls ( n = 8). Grobeta mRNA expression in rabbit lung samples increased with oxygen exposure until day 8, then returned toward baseline levels. This corresponded to previously described elevations in neutrophil number in the lungs. TNFalpha mRNA expression in lung samples was below the limit of detection by RPA and showed no upregulation in hyperoxic lung samples by RT-PCR. TNFalpha activity was assessed in lung lavage ( n = 2 samples per condition per time) using an L929 cell line bioassay and was not increased in hyperoxic animals. The expression of Grobeta mRNA without antecedent or concurrent TNFalpha mRNA expression or activity makes it unlikely that Grobeta in the hyperoxic newborn rabbit lung is elaborated in response to a stimulus by TNFalpha. PMID- 14749939 TI - The oxidant-antioxidant balance in mild asthmatic patients. AB - We investigated the oxidant-antioxidant balance and the effect of inhaled corticosteroids on this balance in mild stable asthmatics. Included in the study were 30 mild asthmatic patients (11 male, 19 female, mean age (year) +/- SD: 35.1 +/- 9.7) and 26 healthy adults (7 male, 19 female, mean age (year) +/- SD: 40.8 +/- 13.3). In all study groups, the peripheral venous blood samples were taken for plasma malonyldialdehyde (MDA), a parameter of lipid peroxidation caused by the oxidants, and erythrocyte superoxide dismutase (SOD), an antioxidant enzyme. The mean plasma MDA level was lower in the asthmatic group (5.7 +/- 1.2 nmol/ml) than in the healthy group (6.3 +/- 1.7 nmol/ml); and the mean erythrocyte SOD level was higher in asthmatic group (1086.4 +/- 247.4 U/gHb) than in the healthy group (1028.0 +/- 230.0 U/gHb). However, there were no significant differences in measurements of both plasma MDA levels and erythrocyte SOD enzyme activities between the groups (respectively, p = 0.1 and p = 0.4). When asthmatic patients were divided into subgroups as "inhaled steroid user" and "no inhaled steroid user", no significant differences were observed in the measurements of either plasma MDA level or erythrocyte SOD enzyme activity between the mentioned subgroups. According to the results of our study, we can say that oxidant antioxidant balance is not significantly affected in mild asthmatics or measurement of plasma level of MDA and erythrocyte SOD enzyme activity is not sensitive to the oxidant-antioxidant balance in mild asthmatics. PMID- 14749940 TI - Low concentration silver nitrate pleurodesis in rabbits: optimal concentration for rapid and complete sclerosing effect. AB - Pleurodesis is a useful therapeutic tool when local treatment of a recurrent malignant pleural effusion or pneumothorax is needed. We have previously demonstrated that the intrapleural injection of 0.5% silver nitrate (SN) produces a significant pleurodesis, while 0.25% SN has no sclerosing effect in a rabbit model. The objective of this study was to determine the minimum concentration of SN needed to induce pleurodesis in our experimental model. One hundred twenty male New Zealand white rabbits received 0.3, 0.4, or 0.5% SN (40 animals per group) in a total volume of 2 mL instilled intrapleurally. These animals were sacrificed 3, 7, 14 or 28 days after the intrapleural injection (n = 10 animals per group), and the pleural spaces were then assessed grossly for evidence of pleurodesis and microscopically for evidence of fibrosis and inflammation. By 28 days, all concentrations of SN had produced a pleurodesis. There was evidence of a gross pleurodesis 7 days post-injection in animals that received 0.5% SN (score of 2.8 +/- 0.2 on a scale of 0-4). After 14 days, significant pleural adhesions were evident in the groups that received 0.4 or 0.5% SN. We conclude that SN concentrations as low as 0.3% can effectively produce a pleurodesis within 28 days of intrapleural injection. However, the precocious pleurodesis development observed 7 days after the intrapleural injection of 0.5% SN suggests that this concentration may be optimal when a fast result is necessary. PMID- 14749941 TI - Gonadotropin releasing hormone antagonists and cryopreservation outcome: a review. AB - REVIEW: The outcome of freeze-thaw cycles with pronuclear stage oocytes or embryos, derived from collecting cycles stimulated with gonadotropin-releasing hormone (GnRH)-antagonists' protocols, was reviewed. CONCLUSION: The viability of cryopreserved pronuclear stage oocytes and embryos, the quality of transferred embryos and the pregnancy rates of the freeze-thaw cycles seem to be satisfactory regardless of the type and dose of GnRH-antagonist. PMID- 14749942 TI - Disseminating myxoid leiomyosarcoma of the uterus. AB - CASE REPORT: A 60-year-old female, para 1, menopausal for last 12 years was admitted with complaints of distention and pain abdomen for last 2 months. There was no history of postmenopausal bleeding. On abdominal examination there was a vague mass in lower abdomen reaching upto umbilicus. On vaginal examination, uterus was eight weeks size and a vague mass was felt around it through all fornices. On exploratory laparotomy uterus was studded with multiple friable hemorrhagic growths perforating from fundus and filling the peritoneal cavity. TREATMENT: Total abdominal hysterectomy with bilateral salpingo-oophorectomy with debulking of tumour and omentectomy was performed followed by postoperative adjuvant radiotherapy. CONCLUSION: Histopathology report confirmed it to be uterine myxoid leiomyosarcoma which is an extremely rare variant of uterine sarcoma with poor prognosis exhibiting malignant biologic behaviour. PMID- 14749944 TI - Mechanism of combined injuries of the posterior cruciate ligament and the posterolateral ligament while wake boarding. AB - BACKGROUND: We encountered a case involving posterior cruciate ligament (PCL) and posterolateral ligament injuries of the knee due to a noncontact injury while wake boarding. The moment of injury was recorded on a home video. CASE REPORT: Based on our analysis of the video, the mechanism of injury was considered to be a combination of varus stress and internal rotation of the lower leg that induced tearing of the posterolateral ligaments. Subsequently, tearing of the PCL in the midsubstance occurred due to increased flexion of the knee. PMID- 14749945 TI - Cerebral fluid edema: an unusual complication of ventriculoperitoneal shunts. AB - INTRODUCTION: A case of accumulation of CSF into the brain parenchyma simulating a brain tumor, secondary to an obstructed ventriculoperitoneal shunt, is presented. Until now, only seven cases of this rare complication have been described. CASE REPORT: Magnetic resonance showed an expansive, low-density intracranial lesion on the right frontal and parietal lobe. This mass was biopsied, but no tumor was found and the diagnosis was brain edema. CONCLUSION: The mistake in the diagnosis was due to the clinical symptoms and to the MR images. PMID- 14749949 TI - Community-acquired and nosocomial pneumonia. AB - Pneumonia is one of the leading causes of morbidity, hospitalization, and mortality in both industrialized and developing countries. In particular, pulmonary infections acquired in the community, and pneumonias arising in the hospital setting, represent a major medical and economic problem and thus a continuous challenge to health care. For the radiologist, it is important to understand that community-acquired pneumonia (CAP) and nosocomial pneumonia (NP) share a number of characteristics, but should, in many respects be regarded as separate entities. CAP and NP arise in different populations, host different spectra of causative pathogens, and pose different challenges to both the clinician and the radiologist. CAP is generally seen in outpatients, is most frequently caused by Streptococcus pneumoniae, Mycoplasma pneumoniae, Haemophilus influenzae, and Chlamydia, and its radiologic diagnosis is relatively straightforward. NP, in contrast, develops in the hospital setting, is commonly caused by gram-negative bacteria, and may generate substantial problems for the radiologist. Overall, both for CAP and NP, imaging is an integral component of the diagnosis, important for classification and differential diagnosis, and helpful for follow-up. PMID- 14749948 TI - Imaging of the postoperative middle ear. AB - The aim of this article is twofold: (a) to present the principles and the indications of surgical treatment of middle ear pathologies; and (b) to review the imaging findings after middle ear surgery, including the normal postoperative aspects and imaging findings in patients presenting with unsatisfactory surgical results or with suspicion of postoperative complications. This review is intentionally restricted to the most common diseases involving the middle ear: chronic otitis media and otosclerosis. In these specific fields of interest, CT and MR imaging play a very important role in the postoperative follow-up and in the work-up of surgical failures and complications. PMID- 14749946 TI - The role of imaging in the staging and treatment planning of primary malignant bone tumors in children. AB - The accurate staging of primary bone tumors in children is critical for treatment planning. Limb salvage operations can now be performed with excellent outcomes in suitable patients. The purpose of this article is to review the current state of imaging techniques and their roles in enabling accurate staging and treatment planning to be performed in pediatric patients with primary bone tumors. PMID- 14749950 TI - Gastrointestinal infection in the immunocompromised (AIDS) patient. AB - In the past two decades acquired immunodeficiency syndrome (AIDS) has become one of the most devastating illnesses in human history. As the epidemic continues to spread increasingly, AIDS patients are no longer confined to a few specialized AIDS hospitals and are now seen in general hospitals and clinics everywhere. Radiologists need to recognize the appearances, to understand how-safely-to care for patients with this disease, and to know enough about the illness to be able to counsel their patients. This article presents a review of current knowledge about the wide range of gastrointestinal hepatic, splenic, biliary, and retroperitoneal manifestations in AIDS, and how the role of modern medical imaging techniques and diagnosis and treatment can be applied. The imaging aspects (conventional double-contrast gastrointestinal studies, ultrasound, CT, and MR) of the diseases of the luminal gastrointestinal tract, liver, spleen, biliary tract, and retroperitoneum will be systematically discussed. Candidiasis, herpes, cytomegalovirus, cryptosporidiosis, histoplasmosis, isosporiasis, salmonellosis, toxoplasmosis, unusual mycobacteria, and viral infections account for the majority of non-neoplastic disorders. PMID- 14749951 TI - Spinal and spinal cord infection. AB - This review article includes infections of the vertebral body, intervertebral disc, ligaments and paravertebral soft tissues, epidural space, meninges and subarachnoid space, and the spinal cord. A wide range of infective organisms may be implicated and the incidence of some, notably tuberculosis, is rising due to increased immunocompromise and other factors. Imaging plays a key role in early diagnosis of these diseases, which may be severe and potentially life threatening. Infection may be acquired by the hematogenous route, by infection from contiguous structures or from direct inoculation. Of available imaging techniques, CT and MRI offer the clinically most valuable methods of evaluating all the structures potentially involved in infection. Although many signs are non specific, indication is given of where appearances raise strong suspicion of infection. The extent of the inflammatory process is well evaluated by imaging, particularly in terms of severity and morbidity of clinical sequelae, which may be severe. Early investigation is mandatory and as MRI presently has the key role in investigation, it should be employed at an early stage of clinical suspicion. PMID- 14749952 TI - Imaging of urinary tract infection in the adult. AB - The diagnosis of urinary tract infection (UTI) in the adult is primarily based on typical patient symptomatology and urinary evaluation for the presence of bacteria and white blood cells. Uncomplicated UTI usually does not require radiological evaluation unless it is recurrent. Imaging should, in general, be reserved for those patients in whom conventional treatment has failed or those who have recurrent or unusually severe symptoms. Patients with conditions predisposing to infection, or complications thereof, such as diabetes mellitus or immunocompromised states, may also benefit from early imaging. If pyonephrosis is suspected, early imaging and possible urgent drainage is also warranted. Intravenous urogram and ultrasound have traditionally been used in the assessment of these patients, allowing detection of calculi, obstruction and incomplete bladder emptying. These imaging techniques, while useful, have limitations in the evaluation of renal inflammation and infection in the adult. Computerised tomography has now become accepted as a more sensitive modality for diagnosis and follow-up of complicated renal tract infection. Contrast-enhanced CT allows different phases of excretion to be studied and can define extent of disease and identify significant complications or obstruction. Nuclear medicine has a limited role in the evaluation of urinary tract infection in adults. Its main role is in the assessment of renal function, often prior to surgery. Magnetic resonance imaging has a limited but increasing role. It is particularly useful in those with iodinated contrast allergies, offering an ionising radiation free alternative in the diagnosis of both medical and surgical diseases of the kidney. PMID- 14749953 TI - Cranial bacterial infection. AB - Early diagnosis of cranial sepsis is mandatory if morbidity is to be avoided. In the case of structural integrity of the skull, haematogenous spread or extension from adjacent structures, especially the sinuses, are the most common sources of infection. Infections may be limited to compartments by the meninges or spread diffusely. Focal disease includes brain abscess as well as subdural and extradural empyaema. A history or signs of sinus disease should always be sought. Tuberculosis, lyme disease and listeriosis may present specific pathological findings. A series of cases is presented to illustrate the role of imaging in infective disease and to draw attention to diagnostic and management points of which radiologists should be aware. PMID- 14749954 TI - Soft tissue and joint infection. AB - Soft tissue infection may be acute or chronic and may be introduced by the haematogenous route, or by inoculation, including surgical infection, or spread from other areas. The spectrum of infecting organism differs in primary infection, in immunodeficiency or when foreign bodies, including prostheses, are present. Bacterial infections are usually more rapid than those due to fungi or atypical organisms. Inflammation usually begins as cellulitis, proceeding through necrosis to cavitation and abscess formation, sometimes complicated by haemorrhage. Imaging, apart from MRI in selected cases, is rarely helpful in the early stages, but early diagnosis of pyomyositis and necrotizing fasciitis is mandatory. When infection is established, US, CT and MRI all have individual value in diagnosis, including biopsy, and in directing therapy, including percutaneous or surgical drainage or debridement. Both MRI and CT are best suited to monitoring progress. Septic arthritis presents as rapidly progressive, destructive arthritis, and early diagnosis is essential to prevent long-term morbidity. Ultrasound offers the best method of detecting early joint effusion and synovial thickening, but aspiration is usually required for diagnosis. In more advanced stages CT is valuable for revealing destructive changes, and MRI for documenting intra-articular changes and detecting inflammation in surrounding bone. PMID- 14749956 TI - Vertebral infection. AB - Vertebral infection represents 2-4% of all cases of osteomyelitis. An increase in the incidence of pyogenic as well as granulomatous spondylitis has been noticed. Early radiological diagnosis is of great importance for prompt treatment and prevention of clinically significant consequences which include neural compromise and late spinal deformities. The most frequent causative pyogenic micro-organisms are gram-negative bacteria especially Staphylococcus aureus. An important form of nonpyogenic granulomatous infection is tuberculous spondylitis which represents the most common form of extrapulmonary tuberculosis. The routes of spinal infection include hematogenous spread, postoperative infections, direct implantation during spinal punctures and spread from a contiguous focus. The role of imaging is an early diagnosis, evaluation of extent of infection with special regard to potential neural compromise, differential diagnosis, guidance of diagnostic biopsy, planning of eventual operative procedures and assessment of therapeutic response. Imaging modalities include bone scintigraphy, radiography, CT and MRI. In practice, usually a combination of a sensitive and a specific method is utilised. The only imaging modality which combines high sensitivity with satisfactory specificity is MRI. This is the reason that MRI frequently has become the first imaging modality in clinically suspect spinal infection. The MRI is the method of choice for direct demonstration of extension of infection, especially of eventual epidural abscess or phlegmon and consecutive neural compression. Using MRI monitoring of therapeutic efficiency is possible. PMID- 14749957 TI - Head and neck lesions in the immunocompromised host. AB - Head and neck lesions are encountered in more than 40-50% in patients with immunosuppression (HIV-infected individuals, diabetes mellitus, transplant recipients, patients treated with immunosuppressive drugs or post-radiotherapy). The organs affected are the salivary glands, the lymph nodes, the sinonasal tract, the orbits, the temporal bones, and the pharyngo-laryngeal mucosa. They are mainly involved by granulation tissue, perivascular and perineural inflammation, and neoplasms. In the pediatric population the temporal bone is the most frequent target organ. The most common clinical manifestation of salivary gland involvement is non-specific bilateral painless enlargement of the parotid gland and xerostomia. Lymphoepithelial cyst, sialosis, and lymphoma may be present. The high rate of salivary gland involvement is probably related to the presence of the human immunodeficiency virus within the saliva. Surgery, sclerotherapy by doxycycline, or low-dose radiotherapy may prevent further growth of tumoral lesions. Sinonasal diseases are the most frequent lesions which manifest in immunosuppressed adult patients (66%). They are associated with a trend of decreased survival rate. Invasive aspergillosis is defined by hyphae in the submucosa, and tumor necrosis without host inflammatory response; it may be lethal in 50-80%, especially when the skull base is involved. Computed tomography shows erosion of bone and extrasinonasal extension. The hypointensity of the lesion on T2-weighted images may suggest the diagnosis. Fungal sinus disease can affect 1-10% of transplant recipients and should be suspected when organ rejection is considered. The temporal bone is the most frequent target organ in the immunosuppressed pediatric population due to Eustachian,tube dysfunction. Otomastoiditis, necrotizing external otitis, and otosyphilis are reported. Prompt treatment may avoid lateral sinus thrombosis. Epithelial neoplasms, lymphomas, and Kaposi's sarcoma may also be considered. Kaposi's sarcoma, the most common neoplasm encountered in patients with AIDS, is an indicator of the progression of human immunodeficiency virus infection to AIDS. The lesions are often multifocal at presentation and may affect the skin, the mucosa, and visceral organs. The pathogenesis is unclear, but cytokines and growth factors may play a role. In conclusion, immunosuppressed patients are likely to develop virulent infection with vascular complications. PMID- 14749955 TI - Imaging of gastrointestinal and abdominal tuberculosis. AB - This article discusses the range of manifestations of tuberculosis (TB) of the abdomen, including involvement of the gastrointestinal tract, the peritoneum, mesentery, omentum, abdominal lymph nodes, solid abdominal organs, the genital system and the abdominal aorta. Abdominal TB is a diagnostic challenge, particularly when pulmonary TB is absent. It may mimic many other abdominal diseases, both clinically and radiologically. An early correct diagnosis, however, is important in order to ensure proper treatment and a favorable outcome. Modern imaging is a cornerstone in the early diagnosis of abdominal TB and may prevent unnecessary morbidity and mortality. Generally, CT appears to be the imaging modality of choice in the detection and assessment of abdominal tuberculosis, other than gastrointestinal TB. Barium studies remain superior for demonstrating mucosal intestinal lesions. Ultrasound may be used for follow-up to monitor therapy response. The diagnosis of abdominal TB should be considered if suggestive imaging findings are found in patients with a high index of suspicion. PMID- 14749958 TI - Respiratory infection in the AIDS and immunocompromised patient. AB - Immunocompromised patients are susceptible to infections by a wide range of organisms. In the past several decades, AIDS epidemic, advances in the treatment of cancer, organ transplantation, and immunosuppressive therapy have resulted in large numbers of patients who develop abnormalities in their immune system. Moreover, mildly impaired host immunity as it occurs in chronic debilitating illness, diabetes mellitus, malnutrition, alcoholism, advanced age, prolonged corticosteroid administration, and chronic obstructive lung disease have also been regarded as predisposing factors of pulmonary infections. Imaging plays a crucial role in the detection and management of patients with pulmonary infectious diseases. When pulmonary infection is suspected, knowledge of the varied radiographic manifestations will narrow the differential diagnosis, helping to direct additional diagnostic measures, and serving as an ideal tool for follow-up examinations. Combination of pattern recognition with knowledge of the clinical setting is the best approach to pulmonary infection occurring in the immunocompromised patients. PMID- 14749959 TI - The spectrum of pelvic inflammatory disease. AB - Pelvic infection presents a serious problem for the practicing physician and the radiologist who is called to determine the origin and extent of the inflammatory process. Pelvic infection may mimic clinically other pelvic inflammatory processes and imaging based on the various aspects of it may provide information to distinguish pelvic infection from other pelvic pathological conditions. This article reviews the various imaging modalities involved in the radiological management of pelvic infection, as well as their findings, with special attention to pelvic inflammatory disease (PID). PMID- 14749960 TI - The spectrum of tuberculosis and non-tuberculous mycobacterial infection. AB - The spectrum of radiographic findings in pulmonary infections due to M. tuberculosis (TB) and non-tuberculous mycobacteria (NTM) are being increasingly discussed, primarily as a result of the increasing prevalence of these infections. The NTM organisms are often overlooked as potential infecting organisms, diagnosis is often delayed and radiologists tend not to include NTM in their differential diagnosis. The literature on the imaging appearances of NTM pulmonary infections is now sufficiently large as to identify some features that would help to distinguish an NTM infection from conventional pulmonary tuberculosis, allowing the radiologist at least to raise the possibility of NTM infection. This review article is intended to describe the radiological features, in the immunocompetent host, that accompany TB and NTM pulmonary infections and highlight the imaging features that would favour an NTM as the causative organism. PMID- 14749961 TI - Gastrointestinal parasite infestation. AB - Twenty-five percent of the world's population could be suffering parasitic infestation. Highest prevalence is in underdeveloped agricultural and rural areas in the tropical and subtropical regions. In some areas incidence may reach 90% of the population. In contrast, some major economic projects intended to promote local development have, paradoxically, caused parasitic proliferation, e.g. bilharziasis in Egypt and Sudan and Chagas disease in Brazil. The commonest cosmopolitan gastrointestinal parasite is Entamoeba histolytica. Some intestinal parasite are endemic in temperate climates, e.g. Entrobius vermicularis. The AIDS epidemic has increased the prevalence and severity of parasitic disease, particularly Strongyloides stercolaris. Tropical parasites are seen in Western people who travel to tropical countries. Radiology has acquired a major role in diagnosis and management of gastrointestinal parasite infestations and their complications. PMID- 14749962 TI - Cranial viral infections in the adult. AB - Viral infections of the adult are fortunately rare conditions but may carry serious clinical sequelae. Infection is usually acquired by haematogenous spread during a systemic viral illness and may be acute, subacute or chronic. The pathological basis of neuronal degeneration and attempt to repair is common to all illnesses and diagnosis is generally made by analysis of the pattern of disease. Magnetic resonance imaging is now the mainstay of imaging diagnosis. Acute infections include encephalitis due to a wide range of infecting agents and outcome depends on the severity of the acute episode. Subacute and chronic infections, including HIV encephalopathy, most often produce a progressive leucoencephalopathy and ultimately cerebral atrophy. Additionally, disease may also be immune mediated, that most closely associated with viral infection being acute disseminated encephalomyelitis, which is usually a monophasic illness. Finally, prion diseases are characterised by long incubation period and progressive course, leading to death. PMID- 14749963 TI - Infection of the appendicular skeleton. AB - In relation to the clinical course, infection in bone can be divided into acute, subacute and chronic osteomyelitis. The diagnosis of acute osteomyelitis is often challenging but can best be made by correlating radiography, bone scintigraphy and MRI with clinical information. Radiography should routinely be supplemented by sonography in the newborns and infants, if applicable. Brodie's abscess, which is clinically a subacute form of osteomyelitis, is best diagnosed by the combination of radiography and MRI. Chronic osteomyelitis is divided into primary haematogenous forms and exogenous, mostly post-traumatic, osteomyelitis. In the majority of patients, post-traumatic osteomyelitis is a clinical diagnosis; however, in a number of patients only the correlation of clinical findings, blood tests and imaging reveals the correct diagnosis. Often, MRI and scintigraphic methods, such as scanning with labeled leucocytes, together establish the diagnosis. Chronic recurrent multifocal osteomyelitis may mimic bacterial osteomyelitis but is a distinct disease probably associated with the SAPHO syndrome. PMID- 14749964 TI - Auxarthrol A and auxarthrol B: two new tetrahydoanthraquinones from Auxarthron umbrinum. AB - Two new 1,2,3,4-tetrahydroanthraquinones, auxarthrol A (compound 1) and auxarthrol B (2), along with three known pyrrolyloctatetraenyl-alpha-pyrone pigments, auxarconjugatin A (3), auxarconjugatin B (4) and rumbrin (5), were isolated from the fungus Auxarthron umbrinum. Structure elucidation of new compounds 1 and 2 was accomplished by spectroscopic data analysis while identification of the known pigments (3-5) was achieved by LC-MS-photodiode array detection. PMID- 14749965 TI - Purification and characterization of a new type of serine carboxypeptidase from Monascus purpureus. AB - Carboxypeptidase produced by Monascus purpureus IFO 4478 was purified to homogeneity. The purified enzyme is a heterodimer with a molecular mass of 132 kDa and consists of two subunits of 64 and 67 kDa. It is an acidic glycoprotein with an isoelectric point of 3.67 and 17.0% carbohydrate content. The optimum pH and temperature were 4.0 and 40 degrees C, respectively. The enzyme was stable between pH 2.0 and 8.0 at 37 degrees C for 1 h, and up to 50 degrees C at pH 5.0 for 15 min. The enzyme was strongly inhibited by piperastatin A, diisopropylfluoride phosphate (DFP), phenylmethylsulfonylfluoride (PMSF), and chymostatin, suggesting that it is a chymotrypsin-like serine carboxypeptidase. Monascus purpureus carboxypeptidase was also strongly inhibited by p chloromercuribenzoic acid (PCMB) but not by ethylenediaminetetraacetic acid (EDTA) and 1,10-phenanthroline, indicating that it requires cysteine residue but not metal ions for activity. Benzyloxycarbonyl- l-tyrosyl- l-glutamic acid (Z-Tyr Glu), among the substrates tested, was the best substrate of the enzyme. The K(m), V(max), K(cat), and K(cat) /K(m) values of the enzyme for Z-Tyr-Glu at pH 4.0 and 37 degrees C were 0.86 mM, 0.917 mM min(-1), 291 s(-1), and 339 mM(-1 )s( 1), respectively. PMID- 14749966 TI - Soft copy versus hard copy reading in digital mammography. AB - The objective of this study was to compare soft copy reading at a mammography work station with hard copy reading of full-field digital mammographic images. Mammograms of 60 patients ( n = 29 malignant, n = 31 benign) performed with full field digital mammography (Senographe 2000D, GE, Buc, France) were evaluated. Reading was performed based on hard copy prints (Scopix, Agfa, Leverkusen, Germany) and on 2 k x 2.5 k high-resolution monitors (Sun Ultra 60, Sun Microsystems, Palo Alto, California, USA). Four readers with different levels of experience in mammography categorized the mammograms according to the BI-RADS classification. The comparative study was performed by four readers, and at least 2 months elapsed between the reading sessions. Postprocessing, of course, was available only at the work station (windowing and leveling, zooming, inversion). Sensitivity, specificity, and positive predictive value were evaluated. Diagnostic accuracy of the evaluation was determined. Sensitivity for malignant lesions in hard copy versus soft copy reading was 97% vs 90%, 97% vs 97%, 93% vs 97%, and 76% vs 76% for the four readers, respectively. Specificity was 52% vs 68%, 58% vs 74%, 65% vs 48%, and 61% vs 68%. Accuracy for the classification of malignant lesions according to the BI-RADS categories showed no difference between hard copy and soft copy reading. Soft copy reading is possible with the available system and enables radiologists to use the advantages of a digital system. PMID- 14749967 TI - Automated recognition of lateral from PA chest radiographs: saving seconds in a PACS environment. AB - Images acquired in a two-view digital chest examination are frequently not electronically distinguishable. As a result the lateral and posterioanterio (PA) images are often improperly positioned on a PACS work station. A series of 1998 chest radiographs (999 lateral, 999 PA or AP) were used to develop a neural network classifier. The images were down-sampled to 16 x 16 matrices, and a feed forward neural network was trained and tested using the "leave-one-out" method. Using five nodes in the hidden layer, the neural network correctly identified 987 of the 999 test cases (98.8%) (average of six runs). The simple architecture and speed of this technique suggests that it would be a useful addition to PACS work station software. The accumulated time saved by correctly positioning the lateral and PA chest images on the work station monitors in accordance with each radiologist's hanging protocols was estimated to be about 1 week of radiologist time per year. PMID- 14749968 TI - Transition from film to electronic media in the first-year medical school gross anatomy lab. AB - For the benefit of the first-year gross anatomy students, we digitized and published on a Web site images that had been collected over a 30-year period. We provided a CD-ROM (compact disk, read-only media) containing the image set in higher quality format to students and faculty. We supplemented basic images with hot topics such as CT angiography, virtual colonography, computer-aided diagnosis, and 3D post-processing. Full motion video and moving JPEG (Joint Photo Expert Group) animations were integrated into the atlas. On the post course questionnaire medical students reported that the images on CD-ROM were helpful during the course and for review prior to examinations. Faculty and medical students used the CD-ROM for problem-based learning sections and facilitator training. The images were clear and easily projected during review sessions and were useful for the small group sessions, where they served as examples of normal anatomy. PMID- 14749969 TI - Flavonoids in translucent bracts of the Himalayan Rheum nobile (Polygonaceae) as ultraviolet shields. AB - UV-absorbing substances were isolated from the translucent bracts of Rheum nobile, which grows in the alpine zone of the eastern Himalayas. Nine kinds of the UV-absorbing substances were found by high performance liquid chromatography (HPLC) and paper chromatography (PC) surveys. All of the five major compounds are flavonoids, and were identified as quercetin 3- O-glucoside, quercetin 3- O galactoside, quercetin 3- O-rutinoside, quercetin 3- O-arabinoside and quercetin 3- O-[6"-(3-hydroxy-3-methylglutaroyl)-glucoside] by UV, 1H and 13C NMR, mass spectra, and acid hydrolysis of the original glycosides, and direct PC and HPLC comparisons with authentic specimens. The four minor compounds were characterised as quercetin itself, quercetin 7- O-glycoside, kaempferol glycoside and feruloyl ester. Of those compounds, quercetin 3- O-[6"-(3-hydroxy-3-methylglutaroyl) glucoside] was found in nature for the first time. The translucent bracts of R. nobile accumulate a substantial quantity of flavonoids (3.3-5 mg per g dry material for the major compounds). Moreover, it was clarified by quantitative HPLC survey that much more of the UV-absorbing substances is present in the bracts than in rosulate leaves. Although the flavonoid compounds have been presumed to be the important UV shields in higher plants, there has been little characterisation of these compounds. In this paper, the UV-absorbing substances of the Himalayan R. nobile were characterised as flavonol glycosides based on quercetin. PMID- 14749972 TI - The autonomic nervous system in systemic sclerosis. A review. AB - The autonomic nervous system is an underestimated target of systemic sclerosis alterations. In this review we analyzed the major manifestations of its involvement, reconsidering the main theories of its pathogenesis. PMID- 14749973 TI - Body composition and hip fracture type in elderly women. AB - Many observations support the view that there are significant differences between patients sustaining trochanteric fractures and those sustaining cervical fractures of the hip. Our aim was to evaluate the association between soft tissue composition (fat and lean compartments) and the type of hip fracture sustained. Of 120 consecutive women affected by their first hip fracture admitted to our rehabilitation hospital 102 were included in this cross-sectional study. Body composition was assessed by DXA. Body fat mass was lower in the women with trochanteric fracture than in those with cervical fracture (difference between groups: 2.86 kg; 95% CI 0.10-5.61 kg; p=0.042). The percentage of fat was 30.75+/ 8.77 (mean+/-SD) versus 34.75+/-7.29 (difference between groups: 4.00; 95% CI 0.84-7.16; p=0.014). In contrast, no meaningful differences in body lean mass were shown between the two groups. Logistic multiple regression showed that fat mass was associated with the type of fracture independently of age, height, weight, time between fracture occurrence and DEXA assessment, comorbidity, number of drugs in use, lean mass and bone mineral content. The logistic regression results were similar when fat percentage was substituted for fat mass. The data show that fat but not lean body mass is associated with the type of hip fracture, contributing to the definition of the differences between patients sustaining cervical or trochanteric fractures. We stress the importance of distinguishing the two types of fracture when clinical or epidemiological studies related to body composition, including those regarding nutrition or physical exercise, are performed. PMID- 14749974 TI - The use of corticosteroid therapy in refractory Kawasaki patients. AB - We retrospectively reviewed four children with Kawasaki disease refractory to the standard therapy. Although three patients had unusual complications, all four responded to intravenous methylprednisolone. This therapy may be considered early in the treatment of selected cases of Kawasaki disease. PMID- 14749975 TI - Reappraisal of cervical spine subluxation in Thai patients with rheumatoid arthritis. AB - Subluxation of the cervical spine is one of a number of devastating complications of rheumatoid arthritis. In spite of this, the features of cervical spine subluxation in Thai patients with rheumatoid arthritis have never previously been studied. We enrolled 134 patients with rheumatoid arthritis who were being followed at the rheumatology clinic, Ramathibodi Hospital, during 1978-2001. Radiological examinations were made in lateral neck flexion, extension and open mouth views. Symptoms of neck pain and the results of relevant neurological examinations were recorded at the time of imaging. Other data on clinical features and treatments since diagnosis were reviewed retrospectively. The overall prevalence of cervical spine subluxation was 68.7%, which can be categorised into anterior (26.9%), posterior (14.9%), lateral (17.2%), vertical (16.4%) atlantoaxial and subaxial subluxation (28.4%). The percentages of cervical subluxation in patients who had suffered from the disease for 1, 5, 10 or more than 10 years were 77.8%, 64.9%, 70% and 64.7%, respectively. None of the patients had neurological deficits. No correlation between neck pain and cervical spine subluxation was established. The number of patients treated with corticosteroids was significantly higher in the subluxation group than in the non subluxation group ( p=0.04). However, no difference in duration of treatment and cumulative dosages of steroids was displayed between the two groups. It was concluded that the prevalence of cervical spine subluxation in Thai patients with rheumatoid arthritis is much higher than the average, even in the early phase of the disease. Hence, radiological examination of the cervical spine should be included in the initial evaluation of Thai RA patients. Corticosteroid use was associated with cervical subluxation, regardless of dose and duration of treatment. The possible explanations are that steroids may directly cause ligament laxity, osteoporosis and decreasing muscle mass, which leads to accelerated subluxation, or that steroid treatments are used in more severe cases which have a higher tendency towards cervical subluxation. PMID- 14749976 TI - Low bone mineral density in professional scuba divers. AB - Scuba diving is associated with a 90% reduction in effective weight and with the loss of a weight-bearing effect on joints. These conditions are very similar to the continuous weightlessness that occurs in spaceflight and bed-rest, which are clearly associated with significant bone mass loss. Here, we studied the bone mineral density (BMD) of 66 professional scuba divers using a dual-photon densitometer, and have depicted a reduction in the BMD in comparison to a matched control group of non-divers. Our results suggest that diving is also an activity where the unloading effect alters bone metabolism, leading to a reduction in BMD. PMID- 14749977 TI - The significance of altered gelatinase expression in the synovium of patient with arthritic effusions. AB - In this study we quantified the levels of matrix metalloproteinase-2 and 9 (MMP-2 and 9) in effusions and serial synovial cultures of patients with arthritis of the knee in order to investigate the correlations between MMP and cell counts in effusions as well as the possible roles of the synovium. In 49 patients with arthritis of the knee (series I) we examined the cell counts and the amounts of MMP-2 and 9 in 51 effusions. In 20 knee samples of series I of patients who received arthroscopy (series II), we examined the amounts of MMP-2 and 9 in effusions and serial synovial organ cultures. We also compared the gene expressions of MMP-2 and 9 and MT1-MMP in serial synovial cultures using RT-PCR. In series I, significantly more proMMP-9 appeared in effusions from the inflammatory group than in the non-inflammatory and hemorrhagic group ( p <0.001). The levels of proMMP-9 correlated with the neutrophil counts in the effusions ( p <0.001). In series II synovial cultures, the activities of latent and activated forms of MMP-2 and 9 in lesional areas were all higher than that in paralesional ones ( p <0.05). In RT-PCR analysis, MMP-2, -9 and membrane type 1 MMP mRNA levels of lesional areas also showed increased expression. Our data suggest that the analysis of MMP-9 indicates the inflammatory condition of the joints and that additional synovectomy may be beneficial for patients with inflammatory synovitis, compared with non-inflammatory and hemorrhagic arthritis. PMID- 14749978 TI - Is the involvement of the distal interphalangeal joint in psoriatic patients related to nail psoriasis? AB - The aim of this study was to investigate the relationship between onychopathy and distal interphalangeal (DIP) joint involvement in psoriatic patients. Twenty-five consecutive unselected, unrelated patients with psoriatic onychopathy and 25 consecutive unselected, unrelated patients with psoriatic arthritis without onychopathy, were enrolled in the study. X-ray films of the hands were taken to identify DIP arthritic involvement and/or bone changes of the distal phalanx, which were categorized into five classes (0: no lesions; 1: tuftal minimal erosions; 2: tuftal bone resorption; 3: tuftal periosteal osteitis; 4: overlap of erosive and osteitic changes). Ten psoriatic patients with onychopathy and 8 without showed DIP arthritis, with no statistical differences in this distribution ( p=0.556). Bone changes of the distal phalanx were found in all 25 psoriatic patients with onychopathy and in 18 without. The distribution of patients in different categories of involvement of the distal phalanx showed that patients without onychopathy were markedly distributed in the categories with no or minimal lesions, whereas patients with onychopathy had structural changes prevailing included in categories with more severe bone changes (osteitis and overlap of erosive and osteitic changes) ( p=0.002). Onychopathic patients with DIP arthritis were older than those without ( p<0.0001) and showed a longer duration of onychopathy ( p<0.0001). Although the occurrence of DIP arthritis seems to depend on the duration of nail involvement, no statistical difference has been found in the distribution of DIP arthritis in psoriatic patients with or without onychopathy. In contrast, a topographical association between bone changes of the distal phalanx and dystrophy of the adjacent nail may be advanced. PMID- 14749979 TI - MRI features of pigmented villonodular synovitis (PVNS). AB - The aim of this study was to characterize the magnetic resonance imaging (MRI) features of PVNS. The radiographs and MR images of 23 pathologically proven cases of PVNS were retrospectively reviewed, with emphasis on MR images. There were 9 males and 14 females, mean age 36 years. Of 23 cases, 9 occurred in the hip, 8 in the knee, 3 in the ankle, 2 in the elbow and 1 in the wrist. Typical MRI findings included variable extent of nodular synovial proliferation, from mild proliferation to extensive masses, joint effusion in all cases, and multiple bony erosions in 15. Owing to the tight joint space, bone involvement was frequently seen in the hip, ankle, elbow and wrist. Although the knee joint had a loose capsule, bone involvement was rarely seen. Hemosiderin is a magnetic material, its deposit on proliferative synovial tissue resulting in a spotty low signal or extensive low signal area within the proliferative synovial masses on T(1)- (T1WI) and T(2)-weighted (T2WI) images, best seen on fast field echo (FFE) sequence MRI images. Fat-suppressed sequences obscured the deposit. This is diagnostic of PVNS. The MRI features of PVNS include variable extent of synovial proliferation, joint effusion and erosion of bone, and in particular the deposit of hemosiderin within the synovial masses. The deposit of hemosiderin, appearing as a low signal area best seen on FFE sequence, is diagnostic for PVNS. PMID- 14749980 TI - Association of TAP2 gene polymorphisms in Chinese patients with rheumatoid arthritis. AB - The aim of this study was to investigate the association between the polymorphism of transporters associated with antigen processing ( TAP1/TAP2) genes and rheumatoid arthritis in Chinese patients. A total of 100 RA patients and 99 healthy control subjects were enrolled. Analyses with polymerase chain reaction (PCR) based restrictions were used to identify the polymorphisms of the TAP1 and TAP2 genes, which were mapped on chromosome 6. There was a significant difference in the distribution of the TAP2 gene codon 565 polymorphism frequency between the RA patients and healthy control subjects ( p<0.001). The odds ratio for the risk of the 'A' allele in RA patients was 1.60 (95% CI: 0.82-2.92). No statistical associations in the distribution of the TAP1 gene polymorphism frequency were found between RA patients and controls. There were some physical links found between TAP1/TAP2 gene polymorphism loci. However, there was no linkage observed from TAP1/TAP2 gene polymorphisms and HLA-DRB1*04 between RA patients and healthy controls. We concluded that the TAP2 gene codon 565 'A' allele was associated with RA in Chinese patients in Taiwan. Individuals possessing the 'A' allele had a higher incidence of RA. A lack of association of TAP1 gene polymorphisms between RA patients and healthy individuals was noted. The results of this study provide genetic evidence that TAP2 gene codon 565 polymorphism may play a role in RA. PMID- 14749981 TI - Destructive joint disease in alpha-mannosidosis. A case report and review of the literature. AB - Storage of oligosaccharides due to a deficiency of alpha-mannosidase can lead to joint destruction in children and young adults. Treating hip destruction with a prosthesis might be successful in some of these patients, although diminished bone quality increases the risk of loosening of the prosthesis. PMID- 14749982 TI - The autoimmune lymphoproliferative syndrome (Canale-Smith) in adulthood. AB - The autoimmune lymphoproliferative syndrome (ALPS) or Canale-Smith syndrome is a recently described clinical entity consisting of chronic, non-malignant lymphadenopathy and hepatosplenomegaly together with hypergammaglobulinemia, positive autoantibodies and/or overt autoimmune diseases. It is caused by a genetic defect in the mechanism of programmed cell death (apoptosis) and is characterized by the presence of double-negative (TCR alpha/beta CD4- CD8-) T lymphocytes (DNT). Although well known in pediatric patients, ALPS is an unusual diagnosis in adults. The oldest reported patient was aged 54. We describe another two adult patients in whom a presenting autoimmune disease led to the diagnosis of ALPS. PMID- 14749984 TI - Keloidal scleroderma. AB - Progressive systemic sclerosis (PSS) can involve the skin diffusely or in a limited way. Rarely cutaneous involvement can assume a hypertrophic form, described in the literature as keloidal scleroderma. The authors describe a PSS patient who presented cutaneous manifestations compatible with the diagnosis of this clinical situation. The clinical features, pathogenic mechanism and therapeutic approach are discussed and a broad review of the literature is given. PMID- 14749983 TI - Successful treatment of refractory adult-onset Still's disease with infliximab. A prospective, non-comparative series of four patients. AB - In this prospective, non-comparative case series, four patients with severe and highly active adult-onset Still's disease (AOSD), refractory to high doses of corticosteroids (which had been combined with methotrexate in three of them) and methotrexate were treated with infliximab (initial dose 3-5 mg/kg, continuing at intervals depending on the patient's individual disease activity). Resolution of their symptoms, which was evident within few days after the first infusion, and a parallel rapid improvement of the acute inflammatory response indices were observed in all. Concomitant corticosteroid treatment was reduced after the first courses of treatment with infliximab, which was well tolerated, and complete disease remission was sustained during a 5-18-month follow-up period. Although further studies to confirm long-term efficacy and safety in larger numbers of patients are needed, we suggest that administration of infliximab with observation for objective improvement is the treatment of choice in cases of AOSD refractory to conventional treatment. PMID- 14749986 TI - Polyarteritis nodosa and hypertrophic obstructive cardiomyopathy. A true association? AB - A 60-year-old man with polyarteritis nodosa under treatment presented with syncope. Echocardiography demonstrated hypertrophic obstructive cardiomyopathy; coronary arteriography revealed normal findings, and Holter monitor showed episodes of non-sustained ventricular tachycardia. This is the first report of hypertrophic obstructive cardiomyopathy developing in a patient with polyarteritis nodosa. Further studies should examine whether a true association exists. PMID- 14749985 TI - Improvement of Erdheim-Chester disease in two patients by sequential treatment with vinblastine and mycophenolate mofetil. AB - Erdheim-Chester disease (ECD) is a rare non-Langerhans' form of histiocytosis with a plethora of different clinical manifestations owing to multiple organ involvement. We report two patients who presented initially with different clinical symptoms. The presenting complaint of the first patient was bone pain, predominantly in the legs, whereas in the other patient the initial symptoms were related to obstruction of both ureters, as in idiopathic retroperitoneal fibrosis. Ultimately, ECD was diagnosed in both patients by the occurrence of both pathognomonic manifestations, the histologic presence of non-Langerhans' histiocytes in bone biopsies, and osteosclerotic lesions of the long bones. Because the extraosseous manifestations progressed and a single application of corticosteroids was ineffective, sequential treatment with vinblastine and mycophenolate mofetil, together with prednisolone, was started. At follow-up respectively 15 and 16 months after the start of treatment a beneficial effect was noted in both patients. These cases illustrate the clinical spectrum of ECD, detail the pathognomonic manifestations of this rare disease, emphasize the need to consider ECD as an uncommon but important differential diagnosis in patients with arthralgias or systemic fibrosis, and give the first evidence for a new treatment option. PMID- 14749987 TI - Parvovirus B19-related chronic monoarthritis: immunohistochemical detection of virus-positive lymphocytes within the synovial tissue compartment: two reported cases. AB - Apart from systemic symptoms of viral infection parvovirus B19, the infectious agent in erythema infectiosum, can lead to mainly self-limited acute and chronic arthropathy. Because mild subclinical features of the disease can be easily overlooked, joint affections might appear as isolated symptoms. We here report two cases of chronic monoarthritic symptoms of unknown origin, where immunohistochemical detection of B19-positive lymphocytic cells in the synovial tissue led to the diagnosis of B19 arthropathy. In conclusion, respective virus diagnostics should be considered even in chronic monosymptomatic arthritic lesions. The pathology of B19 arthropathy seems to be due to direct virus infection of cells within the synovia. PMID- 14749988 TI - Inflammatory myopathy in a patient with cutaneous findings of pityriasis rubra pilaris: a case of Wong's dermatomyositis. AB - We report the case of a 46-year-old woman presenting cutaneous lesions similar to those of pityriasis rubra pilaris, characterized by diffuse scaling erythroderma and palmoplantar keratoderma; skin biopsy showed follicular hyperkeratosis with a perivascular lymphocytic infiltrate. One year later she developed an inflammatory myopathy; on the basis of clinical features, a diagnosis of dermatomyositis was made. Treatment with prednisone and hydroxychloroquine led to complete control of the cutaneous and muscular involvement, which was maintained during prednisone tapering. PMID- 14749989 TI - Tuberculous fasciitis in scleroderma. AB - A 25-year old woman visited the hospital because of a painful swelling for 20 days in the midposterior portion of her right thigh. She had been diagnosed with systemic sclerosis and treated for 10 months. An MRI scan of the right thigh showed diffuse fascial thickening and involved the superficial portion of thigh muscles, which were hyperintense on the T(2)-weighted image. A biopsy of the involved muscles revealed chronic granulomatous inflammation with several acid fast bacilli on a Ziehl-Neelsen stain. We here report a case of tuberculous fasciitis manifest with painful swelling of the midposterior muscles of the right thigh without pulmonary tuberculosis in a patient with scleroderma. PMID- 14749990 TI - CMV enteritis causing ileal perforation in underlying lupus enteritis. AB - We report a case of cytomegalovirus (CMV) enteritis in a 31 year-old -woman with lupus enteritis. In August 2002 the patient complained of severe abdominal pain. An abdominopelvic CT scan at the time showed free air in the peritoneal cavity and wall thickening of the ileal loop. She was diagnosed as having panperitonitis due to an ileal perforation, and underwent an emergency laparotomy. The surgical specimen revealed CMV inclusion bodies in the infarcted lesion. Her symptoms improved following the initiation of ganciclovir therapy. To the best of our knowledge, this is the first report in the English literature of an ileal perforation due to CMV infection in a patient with lupus enteritis. PMID- 14749991 TI - An unusual pattern of arthritis in a child with Kawasaki syndrome. AB - Arthritis is reported in one-third of cases with Kawasaki syndrome. It may have an early or a late onset form. We present a 15-month-old-girl who had been referred with complaints of pain and swelling in her left shoulder. Physical examination revealed bulbar conjunctival injection, erythematous lips and pharynx, strawberry tongue, erythematous rash, edema and erythema of the left shoulder, left knee, right elbow and right wrist, and moderate distress in the left shoulder and left hip. She was diagnosed with Kawasaki syndrome, and intravenous immunoglobulin infusion (IVIG) 2 g/kg and aspirin (100 mg/kg/day) were instituted. The patient had two additional episodes of arthritis involving the hip joint on the 8th day, and the shoulder and metacarpophalangeal (MCP) and interphalangeal (IP) joints of her right hand on the 15th day. Turbid material was aspirated in both instances; Gram and Wright's staining of this material showed many leukocytes but no bacteria. A second dose of IVIG (1 g/kg) was given. At the end of the third week all extremities were painless, with a normal range of motion. Arthritis in our patient was the presenting sign, having a 'septic arthritis mimicking' and 'biphasic' pattern. Although the patient presented with severe and recurrent arthritis, which is significantly correlated with severe multisystem disease and the presence or development of coronary artery aneurysm, the response to IVIG was excellent. PMID- 14749992 TI - Two cases of hypertrophic pachymeningitis associated with myeloperoxidase antineutrophil cytoplasmic autoantibody (MPO-ANCA)-positive pulmonary silicosis in tunnel workers. AB - Two cases of hypertrophic pachymeningitis (HP) associated with pulmonary silicosis in tunnel workers are described. In both cases the myeloperoxidase antineutrophil cytoplasmic autoantibody (MPO-ANCA) was positive. Two patients with pulmonary silicosis developed headache and neurological disturbance, and a diagnosis of HP was made. In both cases the serum CRP level and the MPO-ANCA titer were elevated. Corticosteroid therapy produced a rapid improvement in all the clinical and laboratory parameters. Although an association has been noted between exposure to silica dust and ANCA-associated vasculitis, particularly glomerulonephritis, central nervous system involvement is rare. However, there have been some recent reports of HP cases that were positive for ANCA, and the association between HP and vasculitis has been discussed in the medical literature. HP may be one feature of multiorgan involvement in ANCA-associated disease, and the association between silica dust exposure and HP should be considered, as with other forms of ANCA-associated vasculitis. PMID- 14749993 TI - Lymphadenopathy in adult-onset Still's disease mimicking peripheral T-cell lymphoma. AB - Lymphadenopathy (LAP) that is seen in adult onset Still's disease (AOSD) may be confused with lymphoma. Here we present a patient with AOSD and with LAP that histopathologically mimicked T-cell lymphoma. PMID- 14749994 TI - Transient bone marrow edema syndrome progressing to avascular necrosis of the hip - a case report and review of the literature. AB - Transient bone marrow edema syndrome (TMES) is a rare disease of unknown etiology. Diagnosis is made by exclusion. There is still controversy as to whether TMES is considered to be a reversible form of avascular necrosis (AVN), a disease entity of its own or a form of non-traumatic algodystrophy. We here describe the extremely rare occurrence of three cases of TMES that progressed to AVN. PMID- 14749995 TI - A comparison of bone mineral density in Indians with psoriatic polyarthritis and healthy Indian volunteers. PMID- 14749996 TI - Rheumatic diseases as causes of fever of unknown origin: an update of classic data. PMID- 14749997 TI - A case of adult-onset Still's disease presenting with angioedema. AB - Adult-onset Still's disease (AOSD) is the adult form of Still's disease. In addition to classic symptoms, rarely urticaria may be seen. The author here presents a case of AOSD in which the patient presented with angioedema and urticaria. PMID- 14749998 TI - Involvement of an inconstant bursa under the head of the second metatarsal bone in spondyloarthritis. PMID- 14749999 TI - Calcium pyrophosphate dihydrate deposition disease of the wrist. PMID- 14750000 TI - Interstitial recurrence, with chronic inguinodynia, after Lichtenstein herniorrhaphy. AB - Not long after Lichtenstein and Shulman (1986) introduced their subaponeurotic repair, Nyhus (1989) expressed concern regarding subprosthetic incarceration. Even though interstitial recurrence was not encountered over the subsequent decade or more, one of us (AIG) was referred three cases over the past 3 years. Two men and a woman suffered from chronic inguinodynia 1-6 years following a Lichtenstein procedure for unilateral primary inguinal herniation. A mass, in an unusual location (spigelian line) was palpated in one, the other two required ultrasound studies for diagnosis. All at surgery revealed indirect sacs, and one also had a separate protrusion in the lateral triangle of the groin. Two were repaired laparoscopically, the other using a bilayer connected prosthetic device. Our hypothesis is that this painful complication is now appearing because of recent modifications to the operative technique. An overlay lax dome-shaped prosthesis cannot be relied on to always initially collapse the inguinal canal. Mini-dissection, by limiting exposure, may prevent placement of the keyhole in the mesh close enough to the internal inguinal ring. Studies are under way to determine the validity of these conclusions. PMID- 14750001 TI - Mycelial production, spread and root colonisation by the ectomycorrhizal fungi Hebeloma crustuliniforme and Paxillus involutus under elevated atmospheric CO2. AB - Effects of elevated atmospheric carbon dioxide (CO2) levels on the production and spread of ectomycorrhizal fungal mycelium from colonised Scots pine roots were investigated. Pinus sylvestris (L.) Karst. seedlings inoculated with either Hebeloma crustuliniforme (Bull:Fr.) Quel. or Paxillus involutus (Fr.) Fr. were grown at either ambient (350 ppm) or elevated (700 ppm) levels of CO2. Mycelial production was measured after 6 weeks in pots, and mycelial spread from inoculated seedlings was studied after 4 months growth in perlite in shallow boxes containing uncolonised bait seedlings. Plant and fungal biomass were analysed, as well as carbon and nitrogen content of seedling shoots. Mycelial biomass production by H. crustuliniforme was significantly greater under elevated CO2 (up to a 3-fold increase was observed). Significantly lower concentrations and total amounts of N were found in plants exposed to elevated CO2. PMID- 14750002 TI - Tolerability and effects of two formulations of oral transmucosal fentanyl citrate (OTFC; ACTIQ) in patients with radiation-induced oral mucositis. AB - BACKGROUND: Oral transmucosal fentanyl citrate (OTFC; ACTIQ) incorporates fentanyl into a lozenge allowing drug delivery through the oral mucosa resulting in rapid pain relief. OTFC is effective for breakthrough pain and could be particularly useful in patients with mucositis. METHODS: This randomized, double blind, crossover study assessed two formulations of OTFC for tolerability in 14 patients with radiation-induced mucositis. On four separate days, patients with grade 3 or 4 mucositis received an OTFC unit 45 min before radiation treatment. Two units had a sweetened matrix formulation and two had a compressed powder formulation. One unit of each formulation contained 200 microg fentanyl and one was placebo. Tolerability, mucositis pain, and formulation preference were evaluated. Changes in oral mucosa were recorded. RESULTS: Both formulations of OTFC were well tolerated. There were no significant differences between formulations in tolerability, patient preference, or VAS pain scores. No changes in oral mucosa were noted. Common treatment-related adverse events included a burning sensation in the mouth, nausea, and vomiting. CONCLUSIONS: Both formulations of OTFC are well tolerated. The presence of fentanyl in either the sweetened matrix or the compressed powder did not alter tolerability or safety. The dose of fentanyl tested did not yield analgesia greater than placebo; future studies of OTFC efficacy in mucositis should evaluate higher doses than 200 microg. PMID- 14750003 TI - Electronic leaf wetness duration sensor: why it should be painted. AB - The purpose of this study was to compare and evaluate the performance of electronic leaf wetness duration (LWD) sensors in measuring LWD in a cotton crop canopy when unpainted and painted sensors were used. LWD was measured with flat, printed-circuit wetness sensors, and the data were divided into two periods of 24 days: from 18 December 2001 to 10 January 2002, when the sensors were unpainted, and from 20 January to 13 February 2002, when the sensors were painted with white latex paint (two coats of paint). The data analysis included evaluating the coefficient of variation (CV%) among the six sensors for each day, and the relationship between the measured LWD (mean for the six sensors) and the number of hours with dew point depression under 2 degrees C, used as an indicator of dew presence. The results showed that the painting markedly reduced the CV% values. For the unpainted sensors the CV% was on average 67% against 9% for painted sensors. For the days without rainfall this reduction was greater. Comparing the sensor measurements to another estimator of LWD, in this case the number of hours with dew point depression under 2 degrees C, it was also observed that painting improved not only the precision of the sensors but also their sensitivity, because it increases the ability of the sensor to detect and measure the wetness promoted by small water droplets. PMID- 14750004 TI - Measuring and modelling plant area index in beech stands. AB - For some beech ( Fagus sylvatica L.) stands with different stand densities the plant area index (PAI) was measured by means of a Licor LAI-2000 plant canopy analyser. The stands are located on the slopes of a valley in south-west Germany and had been treated by different types of silvicultural management (heavy shelterwood felling, light shelterwood felling, control plot). The analyser was used (a) to investigate the light conditions on plots of the same thinning regime, (b) to quantify the differences between the different treatments and (c) to obtain absolute values of PAI for interdisciplinary research. PAI was measured at three different phenological stages (leafless, leaf-unfolding and fully leafed season in 2000) and was found to be about 5.2 for the fully developed canopy on the control plots, 3.2 on the light fellings and about 2.0 for the heavy fellings. In the leafless period PAI was between 1.1 (control) and 0.4 (heavy felling). Measurements made in summer 2000 and summer 2002 were compared, and showed an increase of PAI, especially on the thinned plots. Measurements of photosynthetically active radiation (PAR) above and below the canopy in combination with measured PAI were used to apply Beer's Law of radiation extinction to calculate the extinction coefficient k for different sky conditions and for the different growing seasons on the control plots. The extinction coefficient k for the beech stands was found to be between 0.99 and 1.39 in the leafless period, 0.62 to 0.91 during leaf unfolding and between 0.68 and 0.83 in the fully leafed period. Using PAR measurements and the k values obtained, the annual cycle of PAI was modelled inverting Beer's Law. PMID- 14750005 TI - Effects of 24 weeks of whole body vibration training on body composition and muscle strength in untrained females. AB - The aim of this study was to investigate and to compare the effect of 24 weeks "whole body vibration" training and fitness training on body composition and on muscle strength. Forty-eight untrained females (21.3 +/- 2.0 yr) participated in the study. The whole body vibration group (N = 18) performed unloaded static and dynamic exercises on a vibration platform (35 - 40 Hz, 2.5 - 5.0 mm; Power Plate). The fitness group (N = 18) followed a standard cardiovascular (15 - 40 min) and resistance training program including dynamic leg press and leg extension exercises (20 - 8 RM). Both groups trained 3 times weekly. The control group (N = 12) did not participate in any training. Body composition was determined by means of underwater weighing. Additionally 12 skinfolds were assessed. Isometric (0 degrees /s) and isokinetic (50 degrees /s, 100 degrees /s, 150 degrees /s) knee-extensor strength was measured by means of a motor-driven dynamometer (Technogym). Over 24 weeks there were no significant changes (p > 0.05) in weight, in percentage body fat, nor in skinfold thickness in any of the groups. Fat free mass increased significantly in the whole body vibration group (+ 2.2 %) only. A significant strength gain was recorded in the whole body vibration group (24.4 +/- 5.1 %; 5.9 +/- 2.1 %; 8.3 +/- 4.4 %; 7.6 +/- 1.5 %) and in the fitness group (16.5 +/- 1.7 %; 12.0 +/- 2.7 %; 10.4 +/- 2.3 %; 10.2 +/- 1.9 %), at 0 degrees /s, 50 degrees /s, 100 degrees /s and 150 degrees /s respectively. In conclusion, 24 weeks whole body vibration training did not reduce weight, total body fat or subcutaneous fat in previously untrained females. However, whole body vibration training induces a gain in knee-extensor strength combined with a small increase in fat free mass. The gain in strength is comparable to the strength increase following a standard fitness training program consisting of cardiovascular and resistance training. PMID- 14750006 TI - Evidence of exercise-induced O2 arterial desaturation in non-elite sportsmen and sportswomen following high-intensity interval-training. AB - The aim of this study was to investigate the development of exercise-induced hypoxemia (EIH defined as an exercise decrease > 4 % in oxygen arterial saturation, i. e. SaO (2) measured with a portable pulse oximeter) in twelve sportsmen and ten sportswomen (18.5 +/- 0.5 years) who were non-elite and not initially engaged in endurance sport or training. They followed a high-intensity interval-training program to improve V.O (2)max for eight weeks. The training running speeds were set at approximately 140 % V.O (2)max running speed up to 100 % 20-m maximal running speed. Pre- and post-training pulmonary gas exchanges and SaO (2) were measured during an incremental running field-test. After the training period, men and women increased their V.O (2)max (p < 0.001) by 10.0 % and 7.8 %, respectively. Nine subjects (seven men and two women) developed EIH. This phenomenon appeared even in sportsmen with low V.O (2)max from 45 ml x min ( 1) x kg (-1) and seemed to be associated with inadequate hyperventilation induced by training: because only this hypoxemic group showed 1) a decrease in maximal ventilatory equivalent in O (2) (V.E/V.O (2), p < 0.01) although maximal ventilation increased (p < 0.01) with training, i. e. in EIH-subjects the ventilatory response increased less than the metabolic demand after the training program; 2) a significant relationship between SaO (2) at maximal workload and the matched V.E/V.O (2) (p < 0.05, r = 0.67) which strengthened a relative hypoventilation implication in EIH. In conclusion, in this field investigation the significant decrease in the minimum SaO (2) inducing the development of EIH after high-intensity interval-training indicates that changes in training conditions could be accompanied in approximately 40 % non-endurance sportive subjects by alterations in the degree of arterial oxyhemoglobin desaturation developing during exercise. PMID- 14750007 TI - Circadian rhythms in two types of anaerobic cycle leg exercise: force-velocity and 30-s Wingate tests. AB - Previous studies investigating the impact of circadian rhythms on performance during anaerobic cycle leg exercise have yielded conflicting results. The purpose of the present investigation was firstly, to determine the effect of the time of day on anaerobic performance during a force-velocity test on a cycle ergometer (F V) and the Wingate test and secondly, to relate any changes in anaerobic performance to the circadian rhythm in oral temperature. Nineteen subjects volunteered to take part in the study. In a balanced and randomized study design, subjects were measured for maximal power (P (max)) (force-velocity test), peak power (P (peak)) and mean power (P (mean)) (Wingate test) on six separate occasions. These were at 02 : 00, 06 : 00, 10 : 00, 14 : 00, 18 : 00 and 22 : 00 hours on separate days. There was an interval of 28 h between two successive tests. Oral temperature and body mass were measured before each test. Body mass did not vary during the day but a significant time of day effect was observed for the oral temperature with an acrophase at 18 : 22 +/- 00 : 34 hours. A significant circadian rhythm was found for P (max) with an acrophase at 17 : 10 +/- 00 : 52 hours and an amplitude of 7 %. A time-of-day effect was significant for F (0) and V (0). Also a significant circadian rhythm was observed for P (peak) with an acrophase at 17 : 24 +/- 00 : 36 hours and an amplitude of 7.6 % and for P (mean) with an acrophase at 18 : 00 +/- 01 : 01 hours and an amplitude of 11.3 %. The results indicated that oral temperature, P (peak), P (mean) and P (max) varied concomitantly during the day. These results suggest that there was a circadian rhythm in anaerobic performance during cycle tests. The recording of oral temperature allows one to estimate the time of occurrence of maximal and minimal values in the circadian rhythm of anaerobic performance. PMID- 14750008 TI - Hypertrophic cardiomyopathy--sports-related aspects of diagnosis, therapy, and sports eligibility. AB - Hypertrophic cardiomyopathy (HCM) is one of the primary causes of sudden cardiac death in athletes < 35 years of age. The highest risk of sudden cardiac death is associated with syncope, early age, extreme ventricular hypertrophy, ventricular tachycardia, and a family history of sudden death. The relative risk in competitive sports is unknown. Usually, sports eligibility is rejected. However, some athletes with HCM tolerate extreme athletic lifestyles without complications. Sports-related aspects of diagnosis, therapy, and sports eligibility are presented, and discussed. Two case reports are presented: a 20 year-old professional soccer player and a 66-year-old long-distance runner. Athletes with HCM should not participate in most competitive sports with the possible exception of those of low dynamic and low static intensity. Participation in low to moderate athletic activities may be allowed in selected patients without risk factors and > 35 years of age. PMID- 14750009 TI - The contribution of left ventricular mass to maximal oxygen uptake in female college rowers. AB - The purpose of this study was to evaluate the potential relationship between left ventricular systolic and diastolic function and maximal oxygen uptake in female college rowers. We assessed resting left ventricular systolic and diastolic function in 22 female college rowers (mean age, 19 y) utilizing M-mode, two dimensional and Doppler echocardiography and in a multivariate analysis, and investigated the relationship of these indexes with maximal oxygen uptake, measured separately during 6-min rowing ergometer exercise. Among variables, absolute maximal oxygen uptake (V.O (2)max expressed in l/min) had the strongest correlation with distance for the 6 min of rowing ergometer exercise (r = 0.854, p < 0.0001). Among echocardiographic variables, left ventricular mass (LVM ) showed the strongest correlation with absolute V.O (2)max (l/min) (r = 0.836, p < 0.0001). Left ventricular and atrial dimensions also correlated with absolute V.O (2)max (l/min). However, no correlation with absolute V.O (2)max (l/min) was observed in early peak velocity (E), atrial peak velocity (A) and the E/A ratio. Stepwise multiple regression analysis identified only LVM as independent correlates of absolute V.O (2)max (l/min) (p < 0.001). These results indicate that LVM contributes significantly to absolute maximal oxygen uptake associated with the better rowing performance in college female rowers. PMID- 14750010 TI - Relation between plasma lactate concentration and fat oxidation rates over a wide range of exercise intensities. AB - Increasing exercise intensities will induce an increase in glycolytic flux. High glycolytic activity is associated with reduced fat oxidation rates and increased accumulation of lactate. Both lactate and hydrogen ions have been shown to be directly related to the decreased fat oxidation rates. The aim of the present study was to determine whether the exercise intensity at which maximal fat oxidation rates occur coincides with the intensity at which lactate starts to accumulate in plasma. Thirty-three moderately trained endurance athletes performed a graded exercise test to exhaustion on a cycle-ergometer with 35 W increments every three minutes. Expired gas analysis was performed throughout the test and stoichiometric equations were used to calculate fat oxidation rates. The intensity which elicited maximal fat oxidation (Fat (max)) and the intensity at which fat oxidation rates became negligible (Fat (min)) were determined. Blood samples for lactate analysis were collected at the end of each stage of the graded exercise test. The intensity at which lactate concentration increased above baseline (LIAB) and the lactate threshold (LT-D) were determined (D-max method). Fat (max) was located at 63 +/- 9 % V.O (2)max and LIAB at 61 +/- 5 % V.O (2)max and there appeared to be no statistical difference between the two intensities. Fat (max) and LIAB were significantly correlated. Fat (min) and LT-D were also significantly correlated but were located at different intensities (82 +/- 7 and 87 +/- 9 % V.O (2)max respectively). The data of the present study showed that accumulation of lactate in plasma is strongly correlated to the reduction seen in fatty acid oxidation with increasing exercise intensities. The first rise of lactate concentration occurred at the same intensity as the intensity which elicited maximal fat oxidation rates. PMID- 14750011 TI - Fat oxidation in men and women endurance athletes in running and cycling. AB - Recent studies showed that the maximal fat oxidation seems to be different in men and women and that it can be influenced by type and intensity of exercise. Nineteen endurance trained male (V.O (2)peak 61.3 +/- 4.4 ml x kg (-1) x min ( 1)) and 17 female (V.O (2)peak 52.8 +/- 4.5 ml x kg (-1) x min (-1)) athletes were studied over 30 min at 55, 65 and 75 % V.O (2)peak on a treadmill and a cycling ergometer in order to find the intensity and kind of exercise with the highest absolute fat oxidation. For women, normalised (per body weight) fat oxidation was higher at 75 % V.O (2)peak than at 55 % V.O (2)peak for both running (p = 0.02) and cycling (p = 0.01). Women also oxidised a significantly higher percentage of fat with regard to total energy expenditure than men in running (p = 0.02) and cycling (p = 0.004). Normalised carbohydrate oxidation was significantly higher for men at each tested intensity (p < 0.05) and compared to kind of exercise in men (p = 0.006) and women (p = 0.002) in cycling than in running. Men and women showed a significantly higher normalised fat oxidation for running compared to cycling (p = 0.01). Cycling produced in men (p = 0.06) and women (p = 0.001) significantly more lactate than running. In summary, we found at 75 % V.O (2)peak a higher fat oxidation rate than at 65 % V.O (2)peak and 55 % V.O (2)peak for men and women in cycling and running. This is coincident with lactate threshold in men and women in cycling but not in running, where lactate threshold is higher than 75 % V.O (2)peak. PMID- 14750012 TI - No evidence of sustained myocardial injury following an Ironman distance triathlon. AB - We aimed to determine whether an Ironman distance triathlon resulted in sustained myocardial injury detected by electrocardiography, biochemical markers or echocardiographic assessment of left ventricular systolic and diastolic function. Electrocardiograms, blood for analysis of creatine kinase (CK) and its MB fraction, cardiac troponin I (cTnI) and echocardiograms were obtained in 15 male athletes prior to and at a mean of 4.7 days after competing in the Australian Ironman Triathlon. Regional wall motion scores, left ventricular ejection fraction (LVEF) and mitral inflow parameters were determined from the echocardiograms by a blinded investigator. Levels of cTnI were undetectable in all athletes and total CK was mildly elevated in 7/15 athletes prior to the event. Baseline wall motion, ejection fraction and diastolic filling were normal in all athletes. CK levels were increased post-race (p < 0.05) with a mean post race level of 451U/l. Levels of cTnI were undetectable post-race in 14 athletes with a level of 0.9 microg/l recorded in one athlete, although all were within the laboratory's normal range for the assay. Mitral inflow parameters and LVEF did not change post-race and regional wall motion was normal in 14 of 15 athletes. Regional wall motion abnormalities detected in 1 athlete had resolved by 25 days post-race. These findings indicate that ultraendurance exercise does not result in sustained myocardial injury in this group of elite athletes. PMID- 14750013 TI - Body size and V.O2peak: a new perspective? AB - The purpose of this paper was to show how dimensional analysis and biological similarity theory can be used to define and derive extensive (size-dependent) and intensive (size-independent) variables. The method was then used to analyze the peak rate of oxygen consumption (V.O2peak) data of children, adolescents, and adults to determine if size-independent increases in V.O2peak occur during growth and development. The results indicated growth and development is accompanied by a substantial increase in size-independent V.O2peak, which was associated with improvements in size-independent peak heart rate ( HR(peak)). When differences in fat free mass (FFM) were considered, size-independent HR(peak) was similar in boys and girls within each age-group, but was greater in men than in women. In contrast, when differences in FFM were accounted for, size-independent peak oxygen pulse (O2P(peak)) was independent of age, although males tended to have greater values than females at all ages. Because O2P(peak) is proportional to the product of stroke volume ( SV) and arterial mixed-venous oxygen difference (a vO2peak), when differences in FFM were considered, the size-independent nature of O2P(peak) indicated the combined effects changes in SV(peak) and (a-vO(2)peak) played a secondary role in improving aerobic capacity. In conclusion, during normal growth and development improvement in HR(peak), not stroke volume and arterial mixed-venous oxygen difference, appears to increase aerobic capacity. PMID- 14750014 TI - The effects of a sensorimotor training and a strength training on postural stabilisation, maximum isometric contraction and jump performance. AB - Previous studies revealed that adaptations following sensorimotor training, performed to improve functional joint or postural stability, were characterized by improvements in the rate of force development during maximum voluntary isometric contraction. In classical strength training studies using intense loads it has been shown that improvements in rate of force development is mainly due to adaptations in the intramuscular coordination. The purpose of the present study was to compare possible neuromuscular adaptations in two training groups following either sensorimotor or classical strength training over a period of four weeks. Additionally a control group was investigated to contrast the adaptations seen after training. Postural stability, maximum voluntary isometric contraction and performance in squat-jump and in drop-jump were measured before and after training. The results confirmed the positive effects of both training regimen on rate of force development and on maximum strength during maximum voluntary contraction as well as on jump performance, while only the improvements after the strength training were significant. Strength training reduced iMEG, while it was enhanced after sensorimotor training in most testing situations. Strength training had positive effects also on concentric contractions like squat jump. The sensorimotor training improved performance in reactive drop-jump by enhanced neuromuscular activity immediately after ground contact. It is concluded that classical strength training with high loads basically improves the mechanical efficiency of the efferent drive on the motoneurons, whereas sensorimotor training alters the afferent input on the central nervous system. Both adaptations yield to specific effects during force development. PMID- 14750015 TI - Measuring aerobic fitness of Hispanic youth 10 to 12 years of age. AB - Obesity is a major health problem in the U.S., especially for Hispanic youth. Because maximal/peak oxygen consumption (V.O (2)peak) is one predictor of future weight gain in children, valid field-based methods for determining V.O (2)peak in Hispanic children are needed. The purpose of this study was to validate a field based aerobic fitness test, the 20-m shuttle test (20-MST), in Hispanic boys (n = 58) and girls (n = 67), 10 - 12 years old (mean age +/- SD, 10.7 +/- 0.6 y). Measured V.O (2)peak was determined during a maximal, graded treadmill test using the Bruce protocol. The 20-MST was administered per a standard protocol. Maximal speed attained on the 20-MST and age were used to estimate V.O (2)peak. An intraclass coefficient of 0.82 was obtained on 35 students (16 boys; r = 0.85 and 19 girls; r = 0.79) who completed the 20-MST twice, 1-wk apart. Estimated (44.3 ml x kg (-1) x min (-1) ) and measured (45.1 ml x kg (-1) x min (-1)) V.O (2)peak were not significantly different (p = 0.33). The correlation between the two V.O (2)peak parameters was r = 0.62; p < 0.001, the standard error of the estimate (SEE) was 3.91 ml x kg (-1) x min (-1), and 85.5 % of the measured V.O (2)peak values fell within 5.9 ml x kg (-1) x min (-1) of estimated V.O (2)peak. The weight status of the child did not significantly change these results. The 20-MST combined with the Leger et al. equation is a valid method for predicting V.O (2)peak in Hispanic youth. The test can be used to provide valuable information for intervention design and disease prevention. PMID- 14750016 TI - The validity of physiological variables to assess training intensity in kayak athletes. AB - It appears that training benefits are compromised if excessive training is performed at intensities that are either too low or too high. This suggests a need for accurate methods to monitor training intensity. It has been suggested that heart rate (HR) or lactate concentration ([La -]) can be used to accurately monitor training intensity. The purpose of the present study therefore, was to examine whether the relationship between HR, [La -] and intensity determined during a kayak graded exercise test (GXT) remained stable during constant intensity kayak exercise. Sixteen trained kayak paddlers, (22 +/- 4 y, peak V.O (2) = 3.7 +/- 0.9 l x min (-1)) performed a GXT on a wind-braked kayak ergometer. They then performed a 20-min constant-load test on the kayak ergometer at a power output corresponding to their lactate inflection (LI) intensity. Eight subjects also performed a 20-min constant-load test at a power output corresponding to their lactate threshold (LT) intensity. Differences between constant-load and GXT values were determined using one-way ANOVA (p < 0.05). There were no significant differences between values for HR and V.O (2) derived from the GXT and those measured during both constant-load tests. However, while [La -] also provided a valid marker of the LI training intensity (1.8 +/- 0.3 v 2.1 +/- 0.8 mmol x l ( 1)), [La -] did not provide a valid marker of the LT training intensity (3.8 +/- 0.7 v 5.1 +/- 1.4 mmol x l (-1)). These results suggest that HR, but not [La -1], is similar during both a GXT and constant-load exercise at the LT intensity. PMID- 14750017 TI - Influence of therapeutic ultrasound on skeletal muscle regeneration following blunt contusion. AB - Athletic trainers typically use therapeutic ultrasound to treat skeletal muscle contusion injuries. However, the structural outcomes underlying this treatment are not well understood. Our working hypothesis was that following a blunt contusion injury to the gastrocnemius muscle, ultrasound treatment would facilitate recovery, as indicated by changes in several biological markers of skeletal muscle regeneration. Eighty male Wistar rats (three-month) were studied. Following anesthetic administration, each animal received a bilateral contusion injury to the gastrocnemius muscle. Pulsed ultrasound treatment was subsequently initiated six hours post-contusion injury unilaterally on the right gastrocnemius muscle, and ultrasound treatment was continued once daily for seven days. The left (non-ultrasound treated) and right (ultrasound treated) gastrocnemius muscles of 10 animals per group were excised at 1-, 3-, 5-, 7-, 14-, 21-, 28-, and 40-days post-contusion injury. There were no differences in muscle mass, total protein concentration, or fiber cross-sectional area between the right and left gastrocnemius muscles at any post-injury time point examined. Further, when fiber cross-sectional area was normalized to muscle mass, there were no differences. Myonuclear number and cross-sectional area per myonuclei between the right and left gastrocnemius muscles were not different. These results suggest that ultrasound, as administered, does not hasten or improve the regeneration of skeletal muscle following contusion injury. PMID- 14750018 TI - Chronic physical exercise reduces anxiety-like behavior in rats. AB - While many individuals with anxiety disorders receive drug therapy, many do not respond or adversely respond to drugs. An alternative treatment, exercise, has been shown to relieve negative feelings and induce positive shifts in mood. The purpose of this study was to establish an animal model to specifically test the effects of chronic physical exercise on anxiety-related behaviors. Thirty-two male Sprague Dawley rats were divided into two groups: runners (R) and nonrunners (NR). Runners ran on a treadmill for 45 minutes a day, five days a week, for ten weeks at a moderate intensity. Nonrunners remained in their cages in the treadmill room during the running period and were handled for an equal amount of time. After ten weeks of training, two behavioral tests were administered including the elevated plus maze and open field tests. Results comparing R and NR showed higher responses by R in percent open arm time and center square time during the elevated plus maze test, as well as in number of entries into the center, number of rears, and lower fecal boli count during the open field test, p < 0.05. In addition, there were no differences in total activity levels between groups as indicated by similar closed arm entries in the elevated plus maze test and total lines crossed in the open field test. These results indicate that treadmill training reduces anxiety-like behaviors in two animal tests of anxiety, without a significant change in total activity levels. These data are in support of treadmill training as a model to test the anxiolytic effects of exercise. PMID- 14750019 TI - Variable influence of kaempferol and myricetin on in vitro hepatocellular cholesterol biosynthesis. AB - Flavonoids, e. g., quercetin, luteolin, and taxifolin, are known to inhibit hepatocellular cholesterol biosynthesis. Surprisingly, we found that some flavonoids, namely kaempferol and myricetin, are able to considerably stimulate cholesterol biosynthesis, particularly within a low concentration range (between 0.1 and 10 microM), while they show a variable degree of inhibition at higher concentrations. Kaempferol 3-glucoside even stimulated cholesterol synthesis up to 100 microM. In contrast, kaempferol 7-neohesperidoside was ineffective. These effects were more pronounced in rat hepatocytes than in HepG2 cells except for myricetin which acted stimulatory even at high concentrations in HepG2 cells. These opposing effects on cholesterol biosynthesis are exerted in an indirect manner and seem possible though differential modulation of the complex regulation of HMGCoA reductase. PMID- 14750020 TI - Antihyperglycemic effect of andrographolide in streptozotocin-induced diabetic rats. AB - The antihyperglycemic action of andrographolide, an active principle in the leaves of Andrographis paniculata (Burm. f.) Nees, was investigated in streptozotocin-induced diabetic rats (STZ-diabetic rats). Oral treatment of andrographolide decreased the plasma glucose concentrations of STZ-diabetic rats in a dose-dependent manner. Similar treatment with andrographolide also decreased the plasma glucose in normal rats and the maximal effect was more marked than that in STZ-diabetic rats. Andrographolide at the effective dose (1.5 mg/kg) significantly attenuated the increase of plasma glucose induced by an intravenous glucose challenge test in normal rats. In the isolated soleus muscle of STZ diabetic rats, andrographolide enhanced the uptake of radioactive glucose in a concentration-dependent manner. Moreover, the mRNA and protein levels of the subtype 4 form of the glucose transporter (GLUT4) in soleus muscle were increased after repeated intravenous administration of andrographolide in STZ-diabetic rats for 3 days. These results suggest that andrographolide can increase the glucose utilization to lower plasma glucose in diabetic rats lacking insulin. PMID- 14750021 TI - Antispasmodic effect of the essential oil of Plectranthus barbatus and some major constituents on the guinea-pig ileum. AB - The effects of the Plectranthus barbatus essential oil (PBEO) at concentrations ranging form 1 to 300 microg/mL and some major constituents, i. e., alpha-pinene, myrcene and caryophyllene in the ratio and amount found in the oil, were studied on the contractility of the guinea-pig ileum. PBEO decreased the basal tonus of the ileum with a maximal response (R (max); % of K (+)-contraction) of 62.7 +/- 3.8 % compared with 36.6 +/- 4.5 % inhibition achieved with alpha-pinene and 68 +/- 2.6 % with papaverine. The other constituents had only a slight effect. PBEO also blocked the phasic contractions evoked by acetylcholine with an R (max) of 85 +/- 3.6 % compared with 54.4 +/- 3.5 % inhibition induced by alpha-pinene and 12.4 +/- 5.6 % with caryophyllene. The contractions induced by histamine or barium chloride were also decreased in amplitude by PBEO with an R (max) of 94.3 +/- 5.7 % and 100 %, respectively. In addition, PBEO relaxed tissues pre contracted with 60 mM K (+) with an R (max) of 89.7 +/- 2.7 % compared with 55.4 +/- 4.6 % relaxation induced by alpha-pinene. The other major constituents studied had no significant effect. Furthermore, at the higher concentration used, i. e., 300 microg/mL, PBEO decreased the maximal response of calcium chloride (10 ( - 7) to 10 ( - 3) M) induced contraction in depolarized tissues by 29.1 +/- 9.53 % (p < 0.05). In addition, the component of the contraction elicited by a single dose of carbachol (CCh; 100 microM) added on nifedipine (10 microM) treated tissues or at very low extracellular calcium concentration were blocked by PBEO (300 microg/mL) in 85.7 +/- 7.8 % (p < 0.05; n = 5) and 81.2 +/- 6.4 % (p < 0.05; n = 4), respectively. These data show that PBEO has intestinal relaxant and antispasmodic activity and suggest that this effect is not related to antagonism in extracellular receptors for neurotransmitters or autacoids and it seems to occur downstream of calcium entry or release from internal stores. The main active principle for its relaxant and spasmolytic activity seems to be alpha pinene. PMID- 14750022 TI - Relaxant effects of flavonoids in isolated guinea pig trachea and their structure activity relationships. AB - The structure-activity relationships between flavonoids and their tracheal relaxant action are little known. In the present study, 26 natural and synthetic flavonoids, divided into the five classes of flavones, flavonols, flavanones, isoflavones, and chalcones, were tested, and their IC (50) values were determined. The IC (50) values of these five classes indicated that flavones were more potent than flavonols. Flavones were also more potent than flavanones suggesting that the presence of a double bond between C-2 and C-3 is important. However, flavones were similar to isoflavones in potency. Chalcones, a class with an open C-ring, appeared to be the least potent among these five classes. Introduction of a hydroxy group at position C-6 of flavones increases their relaxant activities. So does adding a hydroxy group at position C-7 of flavones. It appears that the optimal number of hydroxy groups introduced to the A-ring of flavones is one. As more hydroxy groups are introduced to positions at C-5, C-6, and/or C-7 of flavones, the IC (50) values increase. It seems that flavones or flavonols with a pyrogallol moiety either in the A- or B-ring, respectively, have no activity. It appears that flavonols with ortho-hydroxy groups in the B-ring are more potent than those with meta-hydroxy groups. The activity of 6 hydroxyflavone disappears if the C-6 hydroxy group of the A-ring is methoxylated. If the C-4' hydroxy group of the B ring is methoxylated, the relaxant effect of these flavones is also attenuated or disappears. Therefore, the hydroxy group on either the A- or B-ring of flavones and flavonols being methylated resulted in lower potency of the tracheal relaxant effects. However, when all hydroxy groups on both the A- and B-rings of flavones or flavonols are methoxylated this results in higher potency. Therefore, the influence of methoxylation in flavones may be similar to that in flavonols. However, if the C-3 hydroxy group on the C-ring of flavonols, but not flavones which lack this hydroxy group, is methoxylated, the relaxant effects may increase. Glycosylation of the hydroxy group at position C-7 of flavones or flavanones attenuates the relaxant effects. PMID- 14750023 TI - Cytotoxic activity of coumarins from the fruits of Cnidium monnieri on leukemia cell lines. AB - Cnidii monnieri Fructus [CmF; Cnidium monnieri (L.) Cusson] is used as a tonic agent in traditional Chinese medicine. In a previous Chinese herb-cytotoxicity screening test, the ethanol extract of CmF exhibited strong effects on human leukemia (HL-60), cervical carcinoma (HeLa) and colorectal carcinoma (CoLo 205) cells. Then, the CmF extract was subjected to silica gel column chromatography and recrystallization to give five coumarins: osthol, imperatorin, bergapten, isopimpinellin, and xanthotoxin. Among these compounds, osthol showed the strongest cytotoxic activity on tumor cell lines. The structure-activity relationship established from the results indicated that the prenyl group has an important role in the cytotoxic effects. However, imperatorin showed the highest sensitivity to HL-60 cells and the least cytotoxicity to normal PBMCs. Osthol and imperatorin both caused apoptotic bodies, DNA fragmentation, and enhanced PARP degradation in HL-60 cells by biochemical analysis. These results indicate that osthol and imperatorin can induce apoptosis in HL-60 cells. Therefore, osthol and imperatorin are cytotoxic marker substances in the fruits of Cnidium monnieri. PMID- 14750024 TI - Sauchinone, a lignan from Saururus chinensis, suppresses iNOS expression through the inhibition of transactivation activity of RelA of NF-kappaB. AB - Sauchinone, a known lignan, was isolated from the root of Saururus chinensis as an active principle responsible for inhibiting the production of NO in LPS stimulated RAW264.7 cells by activity-guided fractionation. Sauchinone dose dependently inhibited not only the production of NO, but also the expression of iNOS mRNA and protein in LPS-stimulated RAW 264.7 cells. Furthermore, sauchinone prevented LPS-induced NF-kappaB activation, which is known to play a critical role in iNOS expression, assessed by a reporter assay under the control of NF kappaB. However, an electrophoretic mobility shift assay (EMSA) demonstrated that sauchinone did not suppress the DNA-binding activity of NF-kappaB or the degradation of IkappaB-alpha induced by LPS. Further analysis revealed that transactivation activity of RelA subunit of NF-kappaB was dose-dependently suppressed in the presence of sauchinone. Taken together, our results suggested that sauchinone could inhibit production of NO in LPS-stimulated RAW264.7 cells through the suppression of NF-kappaB by inhibiting transactivation activity of RelA subunit. PMID- 14750025 TI - In vitro anti-inflammatory activity of panduratin A isolated from Kaempferia pandurata in RAW264.7 cells. AB - An active compound identified as panduratin A was isolated from a methanol extract of Kaempferia pandurata (Zingiberaceae). We examined the effect of panduratin A on nitric oxide (NO) and prostaglandin E (2) (PGE (2)) production induced by lipopolysaccharide (LPS) in RAW264.7 cells. Modulations of iNOS and COX-2 enzyme expression were evaluated by Western blotting. Panduratin A strongly inhibited both NO (IC (50): 0.175 microM) and PGE (2) (IC (50): 0.0195 microM) production and suppressed both iNOS and COX-2 enzyme expression without any appreciable cytotoxic effect on RAW264.7 cells in a dose-dependent manner. Panduratin A also suppressed the phosphorylation of inhibitor kappaBalpha (IkappaBalpha) and degradation of IkappaBalpha associated with nuclear factor kappaB (NF-kappaB) activation. Furthermore, panduratin A inhibited LPS-induced NF kappaB transcriptional activity in a dose-dependent manner. These results suggest that panduratin A could exert its inhibitory effects on the production of NO and PGE (2) through the suppression of NF-kappaB activation, indicating its potential for use as an anti-inflammatory agent. PMID- 14750026 TI - Modulation of PAI-1 and tPA activity and thrombolytic effects of corilagin. AB - In this study, Charlton's and Tomihisa's methods were modified to investigate the thrombolytic effect of corilagin from the Chinese herbal plant Phyllanthus urinaria L., as well as its effect on carotid artery patency status. The activity of type 1 plasminogen activator inhibitor (PAI-1) in rat plasma or platelet released substances and tissue-type plasminogen activator (tPA) in rat plasma was assayed by use of a chromogenic substrate. The results showed that corilagin had a dose-dependent thrombolytic effect in rats. 5 mg/kg of corilagin produced a nearly similar reperfusion rate to that of 20000 U/kg of urokinase, whereas it produced a lower reocclusion rate than urokinase. Corilagin significantly inhibited PAI-1 activity in rat plasma or platelet-released substances while it elevated plasma tPA activity, in a concentration-dependent manner. Corilagin, however, had no influence on rabbit platelet aggregation. It is indicated that corilagin inhibited PAI-1 activity and increased tPA activity, and this property of corilagin is assumed to be responsible for the thrombolytic effect. Abbreviations. PO:persistent occlusion CR:cyclic reflow PP:persistent patency PAI 1:type 1 plasminogen activator inhibitor tPA:tissue-type plasminogen activator PBS:phosphate buffer solution IC (50):50 % of inhibitory concentration PRP:platelet-rich plasma ADP:adenosine diphosphate AA:arachidonic acid PAF:platelet-activating factor PMID- 14750027 TI - A deglucosylated metabolite of paeoniflorin of the root of Paeonia lactiflora and its pharmacokinetics in rats. AB - Paeoniflorin is a bioactive monoterpene glucoside in Paeoniae Radix (PR), the roots of Paeonia lactiflora (Ranunculaceae). By oral administration to rats with the decoction of PR, the metabolism and pharmacokinetics of paeoniflorin were investigated in this study. A deglucosylated metabolite of paeoniflorin, paeoniflorgenin (PG), in serum was identified based on HPLC/MS and NMR spectral data. HPLC/UV methods were developed for determining PG in serum and feces suspension. A non-compartment model was used for the calculation of pharmacokinetic parameters. Moreover, the metabolism of paeoniflorin by various types of feces was investigated as well. The paeoniflorin levels in serum were below the detection limit throughout the study. The C (max), t (max), and AUC (0 t) of PG were 8.0 microg/mL, 10 min and 487.0 microg min/mL, respectively. Paeoniflorin was found to be hydrolyzed into PG through incubation with feces of rabbit, rat, pig or human. Similar profiles of PG were shown for various types of feces, except for rabbit. In conclusion, paeoniflorin was not absorbed per se, whereas its aglycone paeoniflorgenin was absorbable and circulating in the bloodstream. Rat and pig are appropriate models for investigating the metabolism and pharmacokinetics of paeoniflorin. PMID- 14750028 TI - Involvement of reactive oxygen species, but not mitochondrial permeability transition in the apoptotic induction of human SK-Hep-1 hepatoma cells by shikonin. AB - Shikonin has been demonstrated to exhibit anti-cancer activity, but the underlying mechanisms are poorly understood. In this report, we showed that the administration of shikonin could result in the induction of apoptotic cell death of human hepatoma cell line, SK-Hep-1. As evident by the flow-cytometric studies, shikonin has the capability of generating increased amounts of intracellular reactive oxygen species (ROS) during the early stage of this apoptotic process (ca. one-hour), and subsequently accompanied by the dissipation of mitochondrial transmembrane potential (deltapsi (m)) at 3 hours. Further studies indicated that this apoptotic process could effectively be protected by the pretreatment of shikonin-treated cells with glutathione (GSH) and N-acetylcysteine (NAC), a precursor of GSH, but not by cyclosporin A (CyA), an inhibitor of mitochondrial permeability transition (MPT) pore. These data further proved that ROS-mediated oxidative stress was the pivotal element involved in the induction of apoptosis of SK-Hep-1 cells. Taken together, we suggest that shikonin-induced apoptosis of SK-Hep-1 cells proceeds by an oxidative stress-mediated pathway. PMID- 14750029 TI - Effects of selected flavonoids and caffeic acid derivatives on hypoxanthine xanthine oxidase-induced toxicity in cultivated human cells. AB - Tissue damage as a result of oxygen radical generation may be involved in the pathogenesis of different diseases, carcinogenesis, aging and cell death. The inhibition of the proliferation rate of the immortalised human cell line ECV 304 after oxidant damage by oxygen radicals generated in a hypoxanthine-xanthine oxidase system and the protection provided by some selected flavone and flavonol glycosides as well as by caffeic acid and its derivatives was determined. The cytotoxicity of the reactive oxygen species was differentially influenced by selected flavonoids and seems to be determined by the pattern of substitution and by their lipophilicity. Apigenin and quercetin demonstrated the strongest effect on the inhibition of hypoxanthine-xanthine oxidase-induced toxicity (50 % restitution of the cells at a concentration of 0.36 microM and 3.1 microM, respectively). The beneficial effect of the flavonol glycosides rutin and hyperoside was weak, whereas flavone glycosides such as diosmin showed a better effect of protection. PMID- 14750030 TI - Antifungal chromans inhibiting the mitochondrial respiratory chain of pea seeds and new xanthones from Calophyllum caledonicum. AB - Two new xanthones, caledonixanthone M 1 and caloxanthone L 2, and one new acid, caledonic acid 6 were isolated from the hexane-soluble extract of the stem bark of Calophyllum caledonicum. In the course of this phytochemical study, seven other known compounds - calothwaitesixanthone, calozeyloxanthone, allanxanthone, isoapetalic acid 3, calolongic acid 4, apetalic acid 5 and isocalolongic acid 7 - were isolated. Their antifungal activity against the growth of the human pathogenic fungus Aspergillus fumigatus was then investigated. The results indicated that the crude extract, calolongic acid 4 and isocalolongic acid 7 exhibited strong inhibitory effects with MIC (80) values of 8, 4, 2 microg/mL, respectively. Besides, calolongic acid 4, its lactone derivative 4a and isocalolongic acid 7 markedly reduced the respiration of pea seed mitochondria. PMID- 14750031 TI - allo-aromadendrane- and picrotoxane-type sesquiterpenes from Dendrobium moniliforme. AB - Eight sesquiterpenes, four stilbene derivatives along with two lignans have been isolated from lipophilic fraction of stems of Dendrobium moniliforme (Orchidaceae). Among the eight sesquiterpenes, one allo-aromadendrane-type and four picrotoxane-type sesquiterpenes were identified to be new compounds, and alpha-dihydropicrotoxinin was isolated as a new natural product. Their structures were established on the basis of spectroscopic analysis. PMID- 14750032 TI - Anti-HIV-1 protostane triterpenes and digeranylbenzophenone from trunk bark and stems of Garcinia speciosa. AB - Three new protostanes, garciosaterpenes A ( 1), B ( 2) and C ( 3), together with a new digeranylbenzophenone, garciosaphenone A ( 4) were isolated from the ethyl acetate fractions obtained from the crude methanol extracts of the trunk bark and stems of Garcinia speciosa. The structures were elucidated by spectroscopic methods and chemical reactions. Compounds 1 and 3 showed significant inhibitory activities (IC (50) 15.5 and 12.2 microg/mL, respectively) against HIV-1 reverse transcriptase and in the syncytium assay (EC (50) 5.8 microg/mL with TI 3.4 and 37.0 microg/mL with TI 1.9, respectively). Compound 4 was active in HIV-1 RT assay (IC (50) 23.9 microg/mL), but toxic in the syncytium assay. This work represents the first report on the anti-HIV-1 activities of the protostane triterpenes. PMID- 14750034 TI - Demethylbellidifolin inhibits adhesion of monocytes to endothelial cells via reduction of tumor necrosis factor alpha and endogenous nitric oxide synthase inhibitor level. AB - The effect of demethylbellidifolin (DMB), a major compound of Swertia davidi Franch, on the adhesion of monocytes to endothelial cells induced by oxidized low density lipoprotein (ox-LDL) was studied. Adhesion of monocytes to endothelial cells was induced by treatment with ox-LDL (100 microg/mL) for 48 h. Levels of tumor necrosis factor-alpha (TNF-alpha) and asymmetric dimethylarginine (ADMA, an endogenous inhibitor of NOS) in conditioned medium and the activity of dimethylarginine dimethylaminohydrolase (DDAH) in endothelial cells were measured. DMB (3 or 10 micromol/L) significantly inhibited the adhesion of monocytes to endothelial cells, attenuated an increase in levels of TNF-alpha and ADMA, and a decrease in the activity of DDAH by ox-LDL. The present results suggest that DMB inhibits the increased adhesion of monocytes to endothelial cells induced by ox-LDL, and that the effect of DMB is related to reduction of the ADMA concentration via reduction of TNF-alpha production in cultured endothelial cells treated with ox-LDL. PMID- 14750033 TI - Inhibitory effects of manassantin A and B isolated from the roots of Saururus chinensis on PMA-induced ICAM-1 expression. AB - Cell adhesion inhibitors were isolated from the methanol extract of Saururus chinensis roots by bioactivity-guided fractionation. The active compounds were identified as manassantin A ( 1) and B ( 2), dineolignan compounds. Compounds 1 and 2 inhibited PMA-induced ICAM-1/LFA-1-mediated homotypic aggregation of the HL 60 cells without cytotoxicity with MIC values of 1.0 and 5.5 nM, respectively. Even though 1 and 2 did not affect the adhesion of ICAM-1 to LFA-1, these compounds inhibited PMA-induced ICAM-1 expression in HL-60 cells in a dose dependent fashion. These results suggest that 1 and 2 inhibit cell aggregation through down-regulation of ICAM-1 expression. PMID- 14750035 TI - Anticomplement and antioxidant activities of new acetylated flavonoid glycosides from Centaurium spicatum. AB - In addition to the three acetylated flavonol glycosides, quercetin 3- O-[(2,3,4 triacetyl-alpha-rhamnopyranosyl)-(1-->6)]-beta-galactopyranoside, quercetin 3- O [(2,3,4-triacetyl-alpha-rhamnopyranosyl)-(1-->6)]-3-acetyl-beta galactopyranoside, and quercetin 3- O-[(2,3,4- triacetyl-alpha-rhamnopyranosyl) (1-->6)]-4-acetyl-beta-galactopyranoside, which have recently been isolated from Centaurium spicatum (L.) Fritsch (Gentianaceae), a new pentaacetylated flavonoid glycoside was isolated from the same plant. Structure elucidation, especially the localization of the acetyl groups, and complete (1)H- and (13)C-NMR assignments, was carried out using one- and two-dimensional NMR methods, including (1)H- and (13)C-NMR, DEPT-135 and DEPT-90, and gradient-assisted experiments such as DQF COSY, TOCSY, HSQC and HMBC. The structure of the new flavonoid glycoside was established as quercetin 3- O-[(2,3,4-triacetyl-alpha-rhamnopyranosyl)-(1-->6)] 3,4-diacetyl-beta-galactopyranoside. The anticomplement and antioxidant activities of these compounds were evaluated. The triacetylated flavonoid glycoside showed the highest activity in the two assays. PMID- 14750036 TI - Diterpenes from the aerial parts of Salvia candelabrum and their protective effects against lipid peroxidation. AB - A methanolic extract of the aerial parts of Salvia candelabrum was subjected to multiple chromatographic separation under the guidance of anti-lipid peroxidation assay. From the most active fractions seven abietane and seco-abietane diterpenes were isolated by preparative TLC purification. Besides candesalvoquinone, candelabroquinone, 12- O-methylcandesalvone B, candesalvone B methyl ester and candelabrone (all reported earlier), the known candesalvone B and the new candesalvolactone were identified. The structures were established by means of mass spectroscopy and advanced 2D NMR methods. All the identified compounds were evaluated for antioxidant activity in enzyme-dependent (IC (50) values 3.49 - 10.42 microM) and enzyme-independent (IC (50) values 1.40 - 13.40 microM) systems of lipid peroxidation. All compounds displayed marked concentration-dependent effects in both tests as compared with those of authentic ascorbic, rosmarinic and caffeic acids. The differences in antioxidant capacities observed in the enzyme-independent system allowed conclusions concerning structure-activity relationships. PMID- 14750037 TI - Chemical composition and antibacterial activity of essential oil of Artemisia iwayomogi. AB - The chemical composition of the essential oil from Artemisia iwayomogi Kitamura was analyzed by means of GC and GC-MS. Eighty-five constituents were identified representing 96.23 % of the total oil. Camphor (19.31 %), 1,8-cineole (19.25 %), borneol (18.96 %), camphene (4.64 %), and beta-caryophyllene (3.46 %) were found to be the major components. Furthermore, the oil exhibited antibacterial activity against six Gram-(+) and six Gram-(-) bacteria in tests using the broth dilution method. PMID- 14750038 TI - Polyacetylenes in hairy roots of a panax hybrid. AB - From the hairy roots of an interspecific hybrid ginseng ( Panax ginseng x P. quinquefolium) known as Pgq, three polyacetylenes were isolated: panaxynol, panaxydol and 1,8-heptadecadiene-3,10-diol. These compounds isolated from the hairy roots were used for quantitative analysis to investigate the polyacetylene production of the hairy roots cultured in Gamborg B5 (B5) and 1/8 Murashige-Skoog (MS) liquid media. Maximum growth of the hairy roots was observed (ca. 5.4 g fresh weight/100 mL flask) at 8 weeks of culture in B5 medium. The highest total content of total polyacetylenes was 0.18 % of dry weight at week 8 when cultured in 1/8 MS medium. In addition, we compared the yields of polyacetylenes and ginsenosides in hairy roots cultured in B5 with those in 1/8 MS media and found the highest yields were obtained in the hairy roots cultured in B5 medium (1.24 mmol/flask polyacetylenes and 4.45 mmol/flask ginsenosides at week 8). PMID- 14750039 TI - Galanthamine pattern in Narcissus confusus plants. AB - Galanthamine is an Amaryllidaceae-type alkaloid with acetylcholinesterase inhibitory activity which is used in the treatment of Alzheimer's disease. The distribution of galanthamine and four other alkaloids in different organs of Narcissus confusus plants, as well as the variations occurring during the ontogenic cycle of this plant species, was studied. The five alkaloids were found to be present in all the organs at every stage, with the exception of haemanthamine in senescent flowers. The highest amount of alkaloids occurred in the bulb at the emerging stage, galanthamine being the most abundant, reaching a concentration of up to 2.5 % referred to dry weight. PMID- 14750040 TI - Quantitative analysis of strychnine and Brucine in Strychnos nux-vomica using 1H NMR. AB - A quantitative analysis using (1)H-NMR (Q-NMR) has been developed for the determination of strychnine and brucine in Strychnos nux-vomica seeds and stems. The advantages of the method are that no reference alkaloids are needed for calibration curves, the quantification could be directly realized on a crude extract, strychnine and brucine could easily be distinguished, an overall profile of the preparation (including non alkaloid compounds) could be directly obtained, and a very significant time-gain could be achieved, in comparison to conventional HPLC methods, for instance. PMID- 14750041 TI - DNA microarray for identification of the herb of dendrobium species from Chinese medicinal formulations. AB - A DNA microarray for detecting processed medicinal Dendrobium species (Herba Dendrobii) was constructed by incorporating the ITS1-5.8S-ITS2 sequences of 16 Dendrobium species on a glass slide. Using fluorescence-labeled ITS2 sequences as probes, distinctive signals were obtained for the five medicinal Dendrobium species listed in the Chinese Pharmacopoeia. The established microarray was able to detect the presence of D. nobile in a Chinese medicinal formulation containing nine herbal components. PMID- 14750042 TI - Atypical antipsychotics and new onset diabetes mellitus. An overview of the literature. AB - During the last few years, several case reports and studies have been published on the potential diabetes mellitus (DM)-inducing effect of some atypical antipsychotics, especially clozapine and olanzapine. The purpose of our study was to evaluate diabetogenic effects of atypical antipsychotics in the literature. In order to give a full-scale overview, both peer-reviewed publications and oral and poster presentations on this subject were screened. We found 27 case reports of new-onset DM for clozapine, 39 for olanzapine, 4 for risperidone, and 3 for quetiapine. Related to the year of introduction of these drugs on the market and the number of treatment days of each drug during the last 6 years in 13 western countries, Brazil, and Japan, the cases show an over-representation of cases related to olanzapine and clozapine. In the majority of cases, risk factors (DM family history, obesity, Negroid ethnicity) were present. Eighty-four percent of the cases arose in patients < 50 years, in contrast to the general population (most cases, > 50 years). Comparative epidemiological studies point in the same direction, with two studies showing no differences between the atypical drugs. Antipsychotic agents are used often for treatment of schizophrenia, a condition that appears to be associated with DM also in untreated subjects. Some antipsychotics appear to induce new-onset diabetes mellitus. In view of the health risks of DM and the predisposition in patients with schizophrenia, it is advised to be cautious with prescription of antipsychotics that are associated with new-onset DM. Especially in predisposed patients (family history of DM, obese, Negroid ethnicity), reticence in this respect is required. Moreover, careful monitoring of weight, BMI, and glucose levels is advised both before these antipsychotics are prescribed and during treatment. PMID- 14750043 TI - Patterns of antipsychotic utilization in a tertiary care psychiatric institution. AB - OBJECTIVE: To perform a retrospective survey of discharge medications at a tertiary care psychiatric facility in an attempt to gain insight into, and perhaps an understanding of, the most recent pattern of antipsychotic utilization in patients with a diagnosis not restricted to schizophrenia. METHODS: This is a retrospective survey that used the Department of Pharmacy's computer database to obtain relevant discharge information on all non-geriatric patients discharged from Riverview Hospital between 1 January and 31 December 2000. The records of 372 patients met the inclusion criteria and formed the database for the survey. RESULTS: The results of this survey revealed a relatively high prevalence of antipsychotic polypharmacy (the use of two antipsychotics). Perhaps surprisingly, the highest rate of antipsychotic polypharmacy was found in individuals diagnosed with schizoaffective disorder (49.3%), followed by schizophrenia (44.7%), bipolar disorder (29.9%), and psychosis not otherwise specified (22.5%). CONCLUSION: Although antipsychotic polypharmacy is not a new phenomenon in schizophrenia, this study is the first to document its employment among other diagnoses. PMID- 14750044 TI - Effects of imipramine, fluvoxamine and depressive mood on autonomic cardiac functioning in major depressive disorder. AB - OBJECTIVE: Diminished HR variability is considered to be associated with depression and the increased risk of cardiovascular disease. The pharmacological effects of antidepressants and depressive mood itself may contribute to alterations in autonomic cardiac functioning, but a limited amount of data is available. We studied the effects of two different types of antidepressant treatments (imipramine and fluvoxamine), in addition to the effect of depressive mood, on the cardiovascular system in depressed patients. METHODS: Depressed inpatients were studied during a drug-free period and after 4 weeks of adequate treatment with imipramine (n = 17) or fluvoxamine (n = 24). Heart rate variability, blood pressure variability, and a baroreflex sensitivity index during supine rest and orthostatic challenge were analyzed by means of spectral techniques to obtain noninvasive parameters of sympathetic and parasympathetic activity. RESULTS: Both imipramine and fluvoxamine reduced sympathetic and parasympathetic activity, although the effects of imipramine were much more pronounced. Severity of depression was positively related to mean levels of heart rate and blood pressure in the total patient group. There was no convincing evidence that these relationships differed between depressed patients treated with imipramine and those treated with fluvoxamine. CONCLUSION: Our findings suggest that alterations in mean heart rate and blood pressure in depressed patients after antidepressant treatment are the result of a combined effect of pharmacological actions of antidepressants and improvement of depressive mood state. The present study did not confirm the relationship between clinical state and cardiovascular variability or baroreflex sensitivity. PMID- 14750045 TI - Combination of clozapine and amisulpride in treatment-resistant schizophrenia- case reports and review of the literature. AB - BACKGROUND: The clinical outcome of patients suffering from schizophrenic psychoses has been considerably improved by atypical antipsychotics like clozapine and amisulpride. In patients whose symptomatology cannot be ameliorated by monotherapy, it might be necessary to combine two atypical antipsychotics. While clozapine interacts with a variety of neurotransmitter receptors, amisulpride predominantly binds with high affinity to D3/D2-dopamine receptors. Combination can be considered if a supplementary dopamine-receptor blockade is desired. METHODS: We report on the therapy of 15 patients using a combination regimen of amisulpride and clozapine. Data were collected from patient records. The case reports document previous treatment attempts, describe the reason for the combination therapy, and determine its effect. RESULTS: Major (six cases) or at least marked (eight cases) improvement of previously treatment-resistant positive and negative symptoms could be achieved by using a mean clozapine dose of 375 mg/day (serum level 0.38 mg/l) and an amisulpride dose of 527 mg/day. Additionally, by reducing the clozapine dose compared to monotherapy by 24 %, a significant reduction of side effects was observed. CONCLUSIONS: The combination of amisulpride with clozapine considerably enriches the therapeutic arsenal in cases of severe schizophrenic psychoses. Additional prospective studies are needed in order to systematically evaluate this new treatment strategy. PMID- 14750046 TI - Natural course and placebo response in short-term, placebo-controlled studies in major depression: a meta-analysis of published and non-published studies. AB - BACKGROUND: This study was conducted to investigate the role of natural course of a major depressive episode in short-term, placebo-controlled studies. METHODS: We analyzed for sustained response all placebo arms and tricyclic arms from all randomized three-arm studies that were conducted in patients with a major depressive episode and were submitted to the Medicine Evaluation Board (1979 1991). The Medicine Evaluation Board is the regulatory authority of the Netherlands. A responder was defined as a patient with at least 50% improvement on the Hamilton Depression Rating Scale as compared to baseline score. A study responder was defined as a patient meeting response criteria at endpoint (Last Observation Carried Forward). A sustained responder was defined as a patient who, after becoming responder, remained a responder until the end of the study. RESULTS: The ITT population incorporated 1989 patients in the tricyclic arm and 2042 patients in the placebo arm. There was a statistically significant difference for study responders: 39.3% in the tricyclic treatment group and 27.9% in the placebo group (difference 11.4; CI (95%): 8.5% 14.3%). However, no significant differences in sustained response patterns were found, with the exception of a significantly higher sustained response rate for initial responders at week 4. CONCLUSIONS: While efficacy of tricyclic antidepressants was confirmed in this meta-analysis, the sustained response patterns of active treatment and placebo did not differ substantially, suggesting that many of the patients included in the studies were close to, or at the end of, their episode. PMID- 14750047 TI - Effects of topiramate in the treatment of alcohol dependence. AB - BACKGROUND: Anticonvulsant drugs have been used in the treatment of alcohol addiction with relatively good results. The purpose of the present study was to evaluate tolerance and safety of topiramate in patients presenting alcohol dependence. METHODS: We studied 24 patients that fulfilled alcohol-dependence criteria (DSM-IV) and presented other psychiatric disorders for which the use of topiramate was indicated. During the 12 weeks of the study, the patients received topiramate (262 mg/day) plus the psychoactive drugs they were taking for the other disorders. Carbohydrate-deficient transferrin (CDT) values and measures of craving and alcohol use were taken every 2 weeks. RESULTS: Baseline rating of amount and frequency of craving and alcohol use decreased significantly by week 2, and CDT values decreased from week 6. Topiramate was well tolerated, and there were only three dropouts due to adverse events. CONCLUSION: Topiramate is safe and well tolerated, and may be beneficial in the treatment of alcohol dependence. A placebo-controlled study would be of interest. PMID- 14750048 TI - Effect of risperidone on prolactinoma--a case report. AB - Risperidone induced galactorrhea and hyperprolactinemia have been reported but its role in the growth of prolactinoma is not yet conclusive, due to extreme rarity of such cases. We describe a woman, suffering from Bipolar Disorder-manic episode, who exhibited prolactinoma while on risperidone therapy. The withdrawal of risperidone resulted in disappearance of prolactinoma though her prolactin level remained elevated along with persistent galactorrhea. The change to olanzapine therapy did not show much change in serum prolactin level and galactorrhea. Ultimately, only adding of bromocriptine resulted in disappearance of symptoms of prolactinemia and normal serum prolactin level was achieved and galactorrhea stopped. Further study is recommended to find out relationship between the growth of prolactinoma and risperidone. PMID- 14750050 TI - [White blood cell count as a predictor of glucose tolerance and insulin sensitivity. The role of inflammation in the pathogenesis of type 2 diabetes mellitus]. AB - BACKGROUND AND OBJECTIVE: Chronic subclinical activation of the immune system is probably involved in the pathogenesis of type 2 diabetes. In the present study we examined whether an association exists between a non-specific inflammatory marker such as white blood count (WBC) and factors relevant to the pathogenesis of type 2 diabetes. PATIENTS AND METHODS: The statistical association between WBC and glucose tolerance, insulin secretion and insulin sensitivity estimated in an oral glucose tolerance test (OGTT, n = 607) or measured during an euglycemic clamp (N = 280) was determined in non-diabetic individuals. RESULTS: WBC was positively correlated with body weight (r = 0.32, p < 0.001) and postprandial (2-hour) blood glucose during the OGTT (r = 0.22, p < 0.001) independent of sex, age and percentage body fat. WBC negatively correlated with insulin sensitivity both measured by the euglycemic clamp (r = -0.23, p < 0.001) and estimated from the OGTT (r = -0.34, p < 0.001). This relationship remained significant upon adjusting for sex, age and percentage body fat. No relationship between WBC and insulin secretion independent of percentage body fat and insulin sensitivity was found (p = 0.95). CONCLUSION: An increase in WBC is associated with deterioration of glucose tolerance. This is mostly explained by reduced insulin sensitivity. These results are compatible with the hypothesis that chronic subclinical inflammation is involved in the pathogenesis of type 2 diabetes. PMID- 14750049 TI - [Precautions for intestinal cancer in the workplace. An initiative for secondary prevention in the BASF joint-stock company]. AB - BACKGROUND AND OBJECTIVE: The annual incidence of colorectal cancer in Germany is estimated at 27000 in men and 30000 in women. If the diagnosis is made early the cure rate is over 90%. Against this background the department of occupational medicine and health protection of the BASF Aktiengesellschaft initiated a study on the potential prevention of colorectal cancer among the staff at its Ludwigshafen site. SUBJECTS AND METHODS: The target group included all 13265 actively working employees aged 45 years or above. Those expressing interest were given a standardized questionnaire concerning risk factors for colorectal cancer and a test for occult fecal blood (FOBT). If the test was positive and/or a positive answer was given to the question on blood in the stool or on a positive family history, coloscopy - to be arranged via the general practitioner - was advised, in line with the recommendations of the German Society of Digestive and Metabolic diseases (Deutsche Gesellschaft fur Verdauungs- und Stoffwechselerkrankungen). RESULTS: At the end of the study 3732 employees (337 women, 3395 men, mean age 52 years) had completed the questionnaire and the FOBT results were available. Coloscopy was recommended to 688 employees, 323 of whom (47%) underwent the investigation. Nine of the subjects already had manifest cancer, six of them in the early stage T1 or T2. Adenomatous polyps were found in an additional 61 and subsequently excised. Cost-benefit considerations at the company level or in the area of the health system, respectively, gave favourable ratios of 1:10 and 1:14. CONCLUSION: It requires considerable expenditure to increase the number of participants in the cancer prevention programme and obtain a consequent clarification of suspicious findings. Health care within a company is a valuable complementation in Germany of medical care provided by general practitioners or specialist, if close cooperation between practitioners in the given region is assured. Initiatives like the one presented here protect people personally affected against pain and distress and are also of economic value. PMID- 14750051 TI - [Thoracic neuroblastoma in a young adult]. AB - HISTORY AND CLINICAL FINDINGS: A 21-year-old patient was admitted to the hospital because of massively enlarged cervical lymph nodes. Additionally, a left-sided facial and brachial edema was visible. Auscultation of the left lung was remarkable for diminished breath sounds. EXAMINATIONS: Diagnostic imaging showed an extensive thoracic tumor and enlarged mediastinal and cervical lymph nodes. The diagnosis of neuroblastoma was established by biopsy. TREATMENT AND CLINICAL COURSE: The patient was treated with a polychemotherapy protocol according to the pediatric neuroblastoma study NB97. Subsequently, the patient underwent partial tumor resection, received two further chemotherapy courses and irradiation of the remaining tumor. Because of residual vital tumor cells, a second surgical tumour reduction followed by high-dose chemotherapy with autologous stem-cell support was performed. Two cycles of high-dose retinoic acid followed. Six months after the end of therapy, the patient is in a good condition despite of the presence of residual tumor. CONCLUSION: Neuroblastoma is a very rare tumor in adult patients. Therapy is multimodal and should follow pediatric guidelines for neuroblastoma treatment. PMID- 14750052 TI - [Photodermatoses--case report]. PMID- 14750056 TI - [Organ trade versus reciprocity model. An ethical evaluation]. AB - We perform an ethical evaluation of two models that promise to solve the increasing shortage of organs for transplantations: firstly, the legalization of organ trade, and, secondly, the so called "Reciprocity Model". Thereby unrestricted respect for the individual human being serves as the ethical standard. We conclude that the Reciprocity Model is ethically much more acceptable that organ trade even if this trade were limited to Europe. In addition, the Reciprocity Model can easily be integrated into the current Eurotransplant system of organ allocation. PMID- 14750053 TI - [Photodermatoses--diagnosis and treatment]. PMID- 14750057 TI - [Must platelet aggregation inhibitors be discontinued before surgery?]. PMID- 14750058 TI - [Distal radius fractures]. PMID- 14750059 TI - [Palmar plating with the locking compression plate for dorsally displaced fractures of the distal radius--first clinical experiences]. AB - Injuries and irritation of extensor tendons are common problems in the treatment of fractures of the distal radius when plating is used via a dorsal approach. By the development of locking compression plates the possibility of palmar plating for dorsally displaced fractures of the distal radius is available. In this study our first clinical experiences using the 3.5 mm radius locking compression plate (LCP) are reported. Between February 2002 and September 2002 24 patients with dorsally displaced fractures of the distal radius were treated using a palmar approach with the LCP and included in a prospective study. The mean age of the patients was 52 years (28-87 years). Six weeks and six months after surgery a clinical assessment was done, the range of motion of the injured wrist was measured, and an X-ray control of the injured joint was performed. The preliminary results demonstrate the option of early functional treatment using the locking compression plate. Most of the patients had a good to excellent range of motion of the injured wrist which resulted in an early return to former activity. There were few intra- and postoperative complications. No irritation of the median nerve, no infection was observed. In one case a screw which was placed intraarticularly was removed prematurely. The palmar locking compression plate has been proven as a safe and effective implant for the treatment of dorsally displaced fractures of the distal radius. PMID- 14750060 TI - [Dorsal double plating for fractures of the distal radius--a biomechanical concept and clinical experience]. AB - This article describes the anatomical and biomechanical rationale for stable internal fixation of distal radius fractures using a dorsal approach. The three column biomechanical model is illustrated. Advances in the understanding of the anatomy, the biomechanical model of the three columns and our clinical experience with dorsal double plating have lead to the development of a new set of precontoured 2.4 mm Titanium plates with the option for head locking screws. PMID- 14750061 TI - [Management of complex distal radius fractures]. AB - Complex injuries of the distal radius can be associated with articular comminution, severe soft tissue injury, large metaphyseal and/or diaphyseal bone defects, carpal ligament tears or its combinations. We recommend a standardized three step approach for the management of these severe injuries: external fixation as an emergency procedure; treatment of soft tissue damage and further diagnostics as needed; definitive adapted operative therapy after thorough analysis of the specific injury. PMID- 14750062 TI - [Radio-radial external fixation in the treatment of distal radius fractures allows for free wrist motion]. AB - INTRODUCTION: Joint bridging external fixation in the treatment of distal radius fractures restores radial length, whereas anatomic reduction of articular fragments is difficult. Immobilisation of the wrist joint is a further disadvantage. An extraarticular hybrid fixation of distal radius fractures was introduced that facilitates fracture reduction, safe retention and allows for free wrist movement. METHODS: 30 consecutive patients with extra- and intraarticular comminuted fractures of the distal radius were treated with an extraarticular radio-radial external fixation employing a modified Ilizarov hybrid fixation technique. The operative procedure is described in detail. A clinical and radiological evaluation was carried out on the first and seventh day as well as 6 weeks postoperatively after implant removal. RESULTS: Anatomic reduction was achieved in 24 cases (80%). On the first postoperative day dorsiflexion was 30%, palmar flexion was 51% and pronation/supination was 60%/70% of the uninjured side. Six weeks postoperatively all fractures united. Wrist motion of the affected side was more than 80% of the unaffected side. Neither extensor tendinitis nor pin loosening in the distal fragment did occur. CONCLUSION: The technique of extraarticular hybrid fixation is safe and reliable in the treatment of comminuted fractures of the distal radius. It allows for early wrist movement in the immediate postoperative period. PMID- 14750063 TI - [Acute CRPS I (morbus sudeck) following distal radial fractures--methods for early diagnosis]. AB - INTRODUCTION: CRPS I represents a frequent complication following distal radial fractures. Early diagnosis may prevent chronification of the disease. However posttraumatic pain, swelling and motor disturbances render the differentiation from normal fracture patients more difficult. The incidence of CRPS I in patients at risk and the diagnostic value of clinical evaluation, radiography and thermography in the early posttraumatic phase are analysed. METHODS: 158 consecutive patients with distal radial fractures were followed-up for 16 weeks after trauma. Apart from a detailed clinical examination 8 and 16 weeks after trauma, thermography and bilateral radiographs of both hands were performed. RESULTS: At the end of the observation period 18 patients (11%) were clinically identified as CRPS I. The severity of the preceding trauma and the chosen therapy did not influence the process of the disease. 16 weeks after trauma easy differentiation between normal fracture patients and CRPS I patients was possible. 8 weeks after distal radial fracture clinical evaluation showed a sensitivity of 78% and a specificity of 94%. Thermography (58%) however and bilateral radiography (33%) revealed a poor sensitivity, respectively. The specificity was high for radiography (91%) and again poor for thermography (66%), respectively. CONCLUSION: The results of the study support the importance of clinical evaluation in the early diagnosis of CRPS I. Plain radiographs facilitate the diagnosis as soon as bony changes develop. PMID- 14750064 TI - [Emergency ultrasound for blunt abdominal trauma--meta-analysis update 2003]. AB - Emergency ultrasound has established itself as a key procedure of primary diagnostic work-up for blunt abdominal and multiple trauma. However, in a systematic review published in 2001 ultrasonography turned out to provide an unexpectedly low sensitivity. We conducted an update of this analysis to investigate if test characteristics will be maintained including recent studies. Prospective trials published between January 1957 and January 2003 were identified using the Medline/Oldmedline, Embase and Cochrane Controlled Trials Register databases. The searching strategy comprised a manual search as well as a search along the world-wide web. Qualitative rating was carried out by two investigators using criteria proposed by the Centre for Evidence-Based Medicine, Oxford. We investigated a composite endpoint (i. e., free fluid and/or organ laceration) as well as the single criteria organ injury and free intraabdominal fluid collections. After calculation of two-by-two-tables, Summary Receiver Operating Characteristics (SROC) and Q* values were determined together with their 95% confidence intervals. The Q* value was proposed as the point of intersection where sensitivity equals specificity. In addition, a random effects model was employed to compute common positive and negative likelihood ratios (LR). By assessing the title and/or abstract, 349 of 957 papers contained potentially valid information for the purpose of this review. A total of 67 studies were deemed eligible, nine of which had to be excluded from meta-analysis because of dual publication. This left 58 trials allocating 16,361 subjects for statistical analysis. Despite a trend towards improved study designs observed during the past decade, the included trials were of average methodological quality. Two-thirds of all investigations fulfilled two or less of the six possible quality criteria. The diagnostic reference standard was applied independently in only 40% of all protocols. With regard to the composite endpoint and the sonographic depiction of free fluid, the Q* value was estimated at 0.91, whereas Q* equaled 0.90 for the detection of organ injury. Q* values subsequently decreased with improving study quality and fell clearly below 0.80 in methodologically proper studies. Accounting for a negative LR of 0.23 (composite endpoint) and an assumed prevalence of 35% of intraabdominal injury, a post-test probability of 11% will remain in case of a negative sonogram. In pediatric trauma, ultrasound showed even worse test characteristics (negative LR = 0.43). Thus, in case of a 35% prevalence, the post-test probability has to estimated at 19%. Emergency ultrasound provides high specificity but insufficient sensitivity to reliably rule out intraabdominal injury. PMID- 14750065 TI - Surgical wound infection surveillance. AB - Measuring the frequency of a defined outcome flaw for a series of patients undergoing operative procedures generates information for performance evaluation. Such data influence decisions to improve care if used responsibly. Wound infection (WI), bacterial invasion of the incision, is the most common infectious complication of surgical care and WI prevention has value because the complication affects economic, patient satisfaction, and patient functional status outcomes. WI frequency, one kind of surgical outcome flaw rate, is traditionally used to judge one aspect of surgical care quality. At the author's institution, global WI surveillance was conducted without interruption for 20 years. Results for 85,260 consecutive inpatient operations performed during the period showed that secular changes in infection rates occurred but were not necessarily caused by surgical care quality decrements. PMID- 14750066 TI - Hospital-acquired infections in the surgical intensive care: epidemiology and prevention. AB - Hospital-acquired infections affect 7% to 10% of hospitalized patients and account for approximately 80,000 deaths per year in the United States. Of all infections acquired in the hospital, more than 20% occur in intensive care unit patients. As the number of ICU beds increases, the proportion of ICU infections is likely to increase. The focus of this paper is to review the epidemiology of hospital-acquired infections that occur in the surgical ICU, particularly ventilator associated pneumonia, catheter-associated urinary tract infection, and catheter-related bloodstream infection, and to discuss ICU-related prevention strategies. By implementing effective preventative measures and maintaining strict surveillance of ICU infections, we hope to affect the associated morbidity, mortality, and cost that our patients and society bare. PMID- 14750067 TI - [The role of the ultracision technique in visceral surgery. Ligature-free operation--an illusion?]. AB - We report on 431 operations in conventional visceral surgery with the general use of the ultracision technique for preparation (thyroid resections n = 356, colonic resections n = 28, local excisions of the gastric wall n = 3, gastrectomy n = 14, anterior resections of the rectum n = 19, abdominoperineal resection of the rectum [Miles' operation] n = 11). Ligatures were only used for truncal blood vessel ligation. A thorough study of the technical possibilities is a prerequisite for the use of the ultracision technique. The general use of the ultracision technique leads to a revolutionary change of the surgical technique and saves a considerable amount of suture material. In combination with the use of bipolar electrocoagulation the ultracision technique makes the operation fast and leads to a major reduction of bleeding. The biggest disadvantage of the ultracision technique in comparison to the conventional surgical technique is the high cost of the device at present. Despite of savings of suture material, swabs and blood transfusions, the costs for the ultracision scissors are still higher due to its single use. PMID- 14750068 TI - [Value of local surgical therapy for rectal cancer. A literature analysis]. AB - BACKGROUND: Local excision of pT(1) rectal cancer is recognized as curative treatment, whereas the indication is not clarified for pT(2)-tumours. In addition the prognostic histological analysis criteria and the influence of adjuvant and neoadjuvant measures are open. METHOD: Starting form the publication date 1990 investigations on the local excision, with and without adjuvant or neoadjuvant therapy, were included in our literature search and analysed if they comprised at least 30 patients and a follow up of at least 24 months. We studied the assessment criteria local recurrence, survival rate, histological grading, and the conditions of resection margins. RESULTS: After complete resection of "low grade" rectal cancers local recurrences are expected for the pT(1)-category of mean 8 % and for pT(2)-tumours of mean 22%. Poor differentiated pT(1)-tumours exhibit after complete resection over 18 % and after incomplete, despite adjuvant radio (chemo)-therapy, up to 26% recurrences. For pT(2) "high grade" tumours in mean 50% local recurrences and for pT(2) "low grade" tumours after complete resection and adjuvant irradiation in mean 6% have to be expected. Adjuvant measures cannot lower the local recurrence rate for pT(2) tumours after incomplete resection under 20%. CONCLUSION: Local excision with clear margins represents for pT(1) "low grade" cancer an adequate therapy. Further prospective investigations are necessary to clarify whether adjuvant radio (chemo)-therapy represents a curative therapeutic option for pT(2) "low grade" tumours after complete resection. PMID- 14750069 TI - [Ultrasound diagnosis of rare retrorectal tumors]. AB - The diagnosis of perirectal masses can be accomplished by computer tomography and magnet resonance imaging if the rectum is contrasted. The accuracy of these methods is > 90 %. But often the assessment of the exact borders with respect to the rectum wall and to the genitalia is difficult. Therefore transrectal 3D ultrasound is of major importance in the preoperative staging and postoperative follow-up of rectum cancer and allows the diagnosis of pararectal findings using the ultrasound-guided, transrectal aspiration. By ultrasound-guided aspiration biopsy we detected five cases of pararectal and especially presacral diseases including one case of endometriosis. The diagnostic algorithm is described. PMID- 14750070 TI - [Laparoscopic therapy of colorectal cancer--wishful thinking or realism?]. AB - The laparoscopic resection of the colon and rectum is established for surgery of benign diseases. The main advantages compared to the open-conventional operation technique are reduced postoperative pain, reduced negative influences on pulmonary and gastrointestinal functions as well as a shorter reconvalescence. Minimally invasive curative resections of colorectal carcinomas are still controversially discussed concerning sufficient radicality and the spread of tumor cells. The given oncological standards of the conventional approach can be kept without restriction when resections of the colon and rectum are performed laparoscopically. Newer comparative studies do not describe raised recurrence or lower survival rates following the minimal-invasive procedure. In the coming years prospective randomized multicenter studies must give proof whether or not the long-term survival is comparable with the conventional approach. PMID- 14750071 TI - [Changes in perioperative treatment for elective colorectal resections in Germany 1991 and 2001/2002]. AB - BACKGROUND: To assess changes in perioperative treatment of patients undergoing elective colorectal resections, surveys were sent to all German surgical departments in 1991 and 2001/2002. METHODS: 1,207 chairmen of departments for general or visceral surgery were asked to answer a survey concerning the principles of perioperative treatment of patients undergoing elective colorectal resection. The results of this questionnaire were compared to a survey that had been performed in 1991. RESULTS: 616 chairmen (51.0%) responded to the survey (1991: 76.4%). In 2001/2002 preoperative parenteral alimentation was utilized routinely in only 10.3% (1991: 40.0%) of all hospitals. Preoperative i.v. pyelography was used only in 24.7% of the hospitals (1991: 79.7%). Intraoperative testing of colorectal anastomoses was more common in 2001/2002 (63.7%) than in 1991 (40.1%). At the same time the incidence of "single-shot"-antibiotic prophylaxis increased from 24.0% to 70.4 %. Orthograde bowel lavage, perioperative antibiotic prophylaxis and postoperative parenteral alimentation were use as often in 2001/2002 as in 1991. Intraperitoneal drains were routinely inserted in most of the surgical departments after left-sided colonic resections (2001/2002: 86.2%; 1991: 88.2%) or rectal resections (2001/2002: 90.5%; 1991: 94.4%). CONCLUSION: During the last decade, perioperative therapy for patients undergoing elective colorectal resection has changed substantially. Most of these changes occurred in the perioperative medical treatment. However, surgical traditions like intraperitoneal drainage are still very frequently utilized. PMID- 14750072 TI - [Angiodysplasia of the colon combined with portal hypertension and recurrent gastrointestinal bleeding]. AB - Angiodysplasia of the colon has been reported as one of the most common sources of lower gastrointestinal bleeding in the elderly whereas it is very rare in younger patients. Apart from this, portal hypertension may also have effects on the colon. There are, however, only a few reports which have investigated the colon in patients with portal hypertension. We report the case of a 16-year-old woman who presented massive bleeding from angiodysplasia of the colon assembled together with portal hypertension due to portal vein thrombosis caused presumably by neonatal umbilical cord infection. In this regard we discuss diagnosis, therapy and prognosis of this disease. PMID- 14750073 TI - Genomewide scan in families with schizophrenia from the founder population of Afrikaners reveals evidence for linkage and uniparental disomy on chromosome 1. AB - We report on our initial genetic linkage studies of schizophrenia in the genetically isolated population of the Afrikaners from South Africa. A 10-cM genomewide scan was performed on 143 small families, 34 of which were informative for linkage. Using both nonparametric and parametric linkage analyses, we obtained evidence for a small number of disease loci on chromosomes 1, 9, and 13. These results suggest that few genes of substantial effect exist for schizophrenia in the Afrikaner population, consistent with our previous genealogical tracing studies. The locus on chromosome 1 reached genomewide significance levels (nonparametric LOD score of 3.30 at marker D1S1612, corresponding to an empirical P value of.012) and represents a novel susceptibility locus for schizophrenia. In addition to providing evidence for linkage for chromosome 1, we also identified a proband with a uniparental disomy (UPD) of the entire chromosome 1. This is the first time a UPD has been described in a patient with schizophrenia, lending further support to involvement of chromosome 1 in schizophrenia susceptibility in the Afrikaners. PMID- 14750074 TI - A new era of antifungal therapy. AB - Invasive fungal infections pose major management problems for clinicians caring for hematopoietic cell transplant patients. Two major fungal genera, Candida and Aspergillus, account for most fungal infections. Rates of systemic Candida infection range from 15% to 25%, mostly in the pre-engraftment period. Prophylaxis by fluconazole has dramatically reduced the frequency of early Candida infections. Caspofungin has recently been shown to offer an excellent alternative to amphotericin B (with less toxicity) or fluconazole (with a broader spectrum) for therapy of systemic Candida infections. Aspergillus infections occur in 15% to 20% of allogeneic hematopoietic cell transplant patients, most frequently in the post-engraftment period; they are associated with a severe diminution of cell-mediated immune responses by graft-versus-host disease and prolonged corticosteroid use. Voriconazole, a recently introduced broad-spectrum azole, has excellent activity against Aspergillus and is generally well tolerated. Voriconazole currently offers the best prospect for success and tolerance as a first-line treatment for aspergillosis. Second-line therapies include lipid formulations of amphotericin B, caspofungin, or intravenous itraconazole. Unfortunately, early initiation of therapy for aspergillosis is frequently not possible because of inaccurate diagnostics. One new diagnostic, the galactomannan assay, has recently been approved, and others are in development; these offer promise for earlier diagnosis without the need for invasive procedures. It is hoped that these new therapies and new diagnostics will usher in a new era of antifungal therapy. PMID- 14750075 TI - Specific donor Vbeta-associated CD4 T-cell responses correlate with severe acute graft-versus-host disease directed to multiple minor histocompatibility antigens. AB - CXB-2/By (CXB-2) recombinant inbred mice express a subset of the minor histocompatibility antigen (miHA) repertoire expressed by C.B10-H2(b)/LiMcdJ (BALB.B) mice. On lethal irradiation and the transplantation of H2(b)-matched C57BL/6 (B6) T cell-depleted bone marrow cells, along with naive unfractionated T cells, both strains succumb to acute graft-versus-host disease (GVHD). Although alloreactive B6 CD4(+) T cells are a necessary source of T-cell help for the B6 CD8(+) component of the GVHD response in both recipient strains, they are capable of mediating severe GVHD by themselves only in BALB.B mice. Previous CD4(+) T cell receptor repertoire analysis demonstrated overlapping oligoclonal Vbeta use between the CD4(+) B6 anti-BALB.B and B6 anti-CXB-2 responses, with indications of additional BALB.B unique T-cell responses (Vbeta2 and Vbeta11). We report here that the more severe B6 anti-BALB.B response is not due to a quantitative difference in the responding cells, because the frequency of alloreactive donor CD4(+) T cells over time was equivalent in the spleens of BALB.B versus CXB-2 recipients. The responses were also similar in the number of infiltrating B6 CD4(+) T cells in the lingual epithelium of the 2 recipients. In contrast, a significantly greater degree of infiltration and injury of BALB.B intestinal epithelium correlated with the increased level of clinical GVHD severity. Of most significance, despite the involvement of at least 11 Vbeta-associated CD4(+) T cell families in the overall B6 anti-BALB.B response, the development of severe GVHD correlated with the presence of Vbeta2- and Vbeta11-positive donor T cells. Transplantation of donor CD4(+) T cells from Vbeta-associated families that were shared between the B6 anti-BALB.B and anti-CXB-2 responses resulted in minimal GVHD potential. These data suggest that severe GVHD across miHA barriers depends on the involvement of a restricted number of potent T-cell specificities and implies that there are only a limited number of corresponding responsible miHAs. PMID- 14750076 TI - Acute graft-versus-host disease and steroid treatment impair CD11c+ and CD123+ dendritic cell reconstitution after allogeneic peripheral blood stem cell transplantation. AB - Human dendritic cells (DC) comprise 2 subsets-plasmacytoid CD123(+) and myeloid CD11c(+) DC-that may have distinct roles in the regulation of immunity after allogeneic hematopoietic stem cell transplantation. In this study, we analyzed the kinetics of CD123(+) DC and CD11c(+) DC reconstitution in 31 patients who underwent transplantation with allogeneic granulocyte colony-stimulating factor mobilized peripheral blood (PB) stem cells from HLA-identical sibling donors after myeloablative conditioning. Lineage marker-negative HLA-DR(+) CD11c(+) CD11c(+) DC and lineage marker-negative HLA-DR(+) CD123(+) CD123(+) DC, as well as monocytes and lymphoid subsets, were enumerated in donor grafts and in the PB of patients at various time points after transplantation. Reconstitution of both CD11c(+) DC and CD123(+) DC to normal levels occurred within 6 to 12 months and was not affected by the diagnosis, preparatory regimen, or graft composition. However, PB CD11c(+) DC and CD123(+) DC counts were significantly reduced in patients with acute GVHD grade II to IV (at 1 and 3 months) and grade I (at 1 month). Patients with chronic GVHD instead showed reduced CD123(+) DC counts only 6 months after transplantation. Moreover, treatment with steroids (>0.1 mg/kg) was significantly associated with reduced PB CD11c(+) DC and CD123(+) DC counts at all time points after transplantation. In multivariate analysis, only acute GVHD affected DC reconstitution early after transplantation. These results will prompt new studies addressing whether DC reconstitution correlates with immunity against infectious agents or with graft-versus-tumor reactions after PB stem cell allotransplantation. PMID- 14750078 TI - Which donor should be chosen for hematopoietic stem cell transplantation among unrelated HLA-A, -B, and -DRB1 genomically identical volunteers? AB - The aim of this study was to identify significant prognostic factors by using unrelated genomically HLA-A, -B and -DRB1-identical donors. Such data could help to choose the best donor. We studied 136 consecutive patients with hematologic malignancies and a median age of 32 years (range, 0-55 years) who received hematopoietic stem cell transplantation. Bone marrow grafts were given to 83 and peripheral blood stem cells to 53 patients. The cumulative incidence of grade II to IV acute graft-versus-host disease (GVHD) was 30% and of chronic GVHD was 54%. At 5 years, the overall transplant-related mortality (TRM) was 34%, and patient survival was 50%. In Cox multivariate analysis, 32 potential risk factors were analyzed. Monoclonal antibody OKT-3 during conditioning was correlated with grade II to IV acute GVHD, chronic GVHD, and TRM. HLA-DP mismatch was associated with poor TRM and poor survival. Cytomegalovirus-seropositive patients with a seronegative donor had a decreased leukemia-free survival. Five-year TRM was 14% with no risk factor, 38% with 1 risk factor, and 87% with 2 risk factors. The 5 year survival was 72%, 48%, and 30% with 0, 1, and 2 risk factors, respectively. We concluded that unrelated hematopoietic stem cell transplantation may be improved if an optimal donor and immunosuppression are chosen. PMID- 14750077 TI - Autologous transplantation for diffuse aggressive non-Hodgkin lymphoma in first relapse or second remission. AB - We evaluated the results of high-dose chemotherapy and autologous hematopoietic stem cell transplantation in patients with diffuse aggressive non-Hodgkin lymphoma (NHL) in first relapse (Rel 1) or second complete remission (CR 2). Data were evaluated from the Autologous Blood and Marrow Transplant Registry on 429 patients with diffuse aggressive NHL who underwent transplantation in Rel 1 or CR 2. Transplantations were performed between 1989 and 1996 and were reported to the Autologous Blood and Marrow Transplant Registry by 93 centers in North and South America. The probability of 3-year survival was 44% (95% confidence interval [CI], 33%-55%). The probability at 3 years of progression-free survival was 31% (95% CI, 27%-36%). Patients who underwent transplantation in CR 2 had a 3-year probability of progression-free survival of 38% (95% CI, 30%-46%) compared with 28% (95% CI, 22%-33%) for those who were not in remission at the time of transplantation (P <.001). In multivariate analysis, chemotherapy resistance, increased lactic dehydrogenase at diagnosis, an interval of <12 months from diagnosis to relapse, age >or=40 years, and use of myeloid growth factors to accelerate posttransplantation bone marrow recovery were adverse predictors of survival. High-dose chemotherapy and autologous hematopoietic stem cell transplantation for patients with diffuse aggressive NHL in CR 2 or Rel 1 resulted in better outcome for patients with chemotherapy-sensitive disease, longer relapse-free intervals, and age <40 years. Exposure to myeloid growth factors to accelerate recovery for recipients of bone marrow grafts may increase the risk of disease progression or death. PMID- 14750079 TI - The role of depletion of dimethyl sulfoxide before autografting: on hematologic recovery, side effects, and toxicity. AB - Cryopreservation of stem cells after collection from peripheral blood or bone marrow for autologous transplantation necessitates protection with dimethyl sulfoxide (DMSO). Unfortunately, DMSO, when infused with the thawed cell suspension, may induce serious complications and side effects. To assess whether depletion of DMSO before autografting affects safety and efficacy, 56 consenting consecutive patients treated with high-dose chemotherapy and autologous blood stem cell transplantation were assigned to obtain either an untreated or DMSO depleted autograft. On the day of transplantation, the cryopreserved cells were thawed and infused to the patient either immediately or after washing 3 times in normal saline supplemented with 6% anticoagulant citrate dextrose solution. Cell count with viability, clonogenic assay, and phenotyping were performed before and after thawing and after washing. Hematologic recovery, side effects, and complications were recorded. The in vitro and clinical data on 56 patients show that the depletion of DMSO in vitro before autografting does not induce a significant loss of cell number, viability, colony-forming unit-granulocyte macrophage activity, or number of CD34(+) cells. Furthermore, it leads to a safe and sustained engraftment. The complications and side effects, as recorded by continuous monitoring, were substantially less; however, the procedure takes 3 to 4 hours of laboratory work per patient. PMID- 14750085 TI - Treatment of diffuse proliferative lupus nephritis: a meta-analysis of randomized controlled trials. AB - BACKGROUND: In this systematic review of randomized controlled trials (RCTs), we assess the benefits and harm of current treatments for diffuse proliferative lupus nephritis (DPLN). METHODS: The Cochrane Controlled Trial Registry, MEDLINE, and EMBASE were searched for RCTs of treatment for DPLN. All available RCTs of patients with biopsy-proven DPLN were included, and data were extracted for overall mortality, end-stage renal disease, doubling of serum creatinine level, relapse, major infection, herpes zoster infection, ovarian failure, malignancy, and bladder toxicity. Treatment effects on these outcomes were summarized as relative risk (RR) with 95% confidence interval (CI) and pooled by using a random effects model. RESULTS: Twenty-five of 920 articles identified were eligible RCTs and were included. The majority compared cyclophosphamide or azathioprine plus steroids versus steroids alone. Cyclophosphamide plus steroids reduced the risk for doubling of serum creatinine level (4 RCTs, 228 patients; RR, 0.59; 95% CI, 0.40 to 0.88) compared with steroids alone, but had no impact on overall mortality (5 RCTs, 226 patients; RR, 0.98; 95% CI, 0.53 to 1.82). However, risk for ovarian failure was increased significantly (3 RCTs, 147 patients; RR, 2.18; 95% CI, 1.10 to 4.34). In studies from the 1970s, azathioprine plus steroids reduced the risk for all-cause mortality compared with steroids alone (3 RCTs, 78 patients; RR, 0.60; 95% CI, 0.36 to 0.99), but had no effect on renal outcomes. Neither therapy was associated with increased risk for major infection. The addition of plasma exchange to these treatments offered no benefit, and information on other agents, including mycophenolate mofetil, was insufficient for analysis. CONCLUSION: Until future RCTs of newer agents are completed, the current use of cyclophosphamide combined with steroids remains the best option to preserve renal function in patients with DPLN. The smallest effective dose and shortest duration of treatment should be used to minimize gonadal toxicity without compromising efficacy. PMID- 14750086 TI - Oculorenal manifestations in systemic autoimmune diseases. AB - Vasculitides form a heterogeneous group of diseases characterized by blood-vessel inflammation and necrosis. They have a wide spectrum of manifestations because of the involvement of arteries and other vessels of various sizes and locations. Early diagnosis and prompt treatment may decrease the morbidity and mortality associated with these disorders. Examination of the eye and kidney should be performed routinely in those diseases. This article reviews the major types of oculorenal manifestations in systemic autoimmune diseases. PMID- 14750087 TI - Apolipoprotein E and renal disease. AB - Apolipoprotein E (ApoE) is a major constituent of plasma lipoproteins with many biological actions of great significance. Beyond the known influence of ApoE polymorphisms on serum lipid profile, the pathogenesis of atherosclerosis, and the development of neurodegenerative disorders, ApoE also has a major role in the pathogenesis and progression of a variety of renal diseases, as well as in the atherosclerotic complications associated with them. Briefly, the polymorphisms of ApoE are major determinants of plasma lipid levels in uremic patients. They may affect the risk for cardiovascular disease in this population, predispose to the development of diabetic nephropathy, influence the severity of certain glomerulopathies, and regulate mesangial and glomerular functions locally in the kidney microenvironment. Finally, certain mutations of the ApoE gene are associated with a recently described nephropathy, termed lipoprotein glomerulopathy. PMID- 14750088 TI - Cardiac calcification in renal patients: what we do and don't know. AB - Cardiovascular (CV) disease is one of the major causes of mortality in patients with renal diseases, with an increased odds ratio of mortality with risk factors as diverse as blood pressure (high or low), cholesterol level (high or low), left ventricular hypertrophy, vascular stiffness, chronic inflammation, and hyperhomocysteinemia. Mainly cross-sectional studies of renal patients showed excess CV calcification (CVC) compared with the general population, but a clear link between calcification and subsequent mortality is tenuous to date. Several factors have been incriminated to explain the increase in CVC in this particular population. Increased duration of dialysis therapy, dyslipidemia, altered calcium phosphorus metabolism, and chronic inflammation have all been associated with increased CVC. However, with the shortage of large, observational, population based, prospective studies tracking these potential risk factors and the pathogenesis of CVC in renal patients not yet sufficiently understood, it is difficult with the present state of knowledge to make robust recommendations about care strategies. The purpose of this review is to examine the 10 available studies of renal patients that have used modern CVC imaging and quantification techniques for clues to likely targets for future interventional studies. PMID- 14750089 TI - Inflammation, endothelial dysfunction, and platelet activation in patients with chronic kidney disease: the chronic renal impairment in Birmingham (CRIB) study. AB - BACKGROUND: Studies in the general population suggest that low-grade inflammation, endothelial dysfunction, and platelet activation are associated with an increased risk of cardiovascular events. METHODS: Markers of inflammation, endothelial dysfunction, and platelet activation were measured in 334 patients with chronic kidney disease (serum creatinine >1.47 mg/dL [>130 micromol/L] at screening) and compared with 2 age- and sex-matched control groups, 1 comprising 92 patients with coronary artery disease and the other comprising 96 apparently healthy individuals with no history of cardiovascular or kidney disease. RESULTS: There was evidence of low-grade inflammation in the chronic renal impairment group compared with healthy controls, with higher concentrations of C-reactive protein (3.70 versus 2.18 mg/L, P < 0.01) and fibrinogen (3.48 versus 2.67 g/L, P < 0.001) and lower serum albumin concentration (41.8 versus 44.0 g/dL [418 versus 440 g/L], P < 0.001). More severe renal impairment was associated with a trend towards higher fibrinogen and lower albumin concentrations (both P < 0.001), although there was no association with higher C-reactive protein level. As compared to healthy controls, plasma von Willebrand factor (142 versus 108 IU/dL, P < 0.001) and soluble P-selectin concentrations (57.0 versus 43.3 ng/mL, P < 0.001) were also higher in the chronic renal impairment group. More severe renal impairment was associated with a trend towards higher levels of von Willebrand factor (P < 0.001) and of soluble P selectin (P < 0.05). CONCLUSION: This cross-sectional analysis demonstrates that chronic kidney disease is associated with low-grade inflammation, endothelial dysfunction, and platelet activation, even among patients with moderate renal impairment. PMID- 14750090 TI - Kidney volume, blood pressure, and albuminuria: findings in an Australian aboriginal community. AB - BACKGROUND: Australian Aborigines are experiencing epidemic proportions of renal disease, marked by albuminuria and, variably, hematuria. They also have high rates of low birth weight, which have been associated with lower kidney volumes and higher blood pressures. The authors evaluated relationships between kidney volume, blood pressure, albuminuria, and hematuria in 1 homogeneous group. METHODS: Forty-three percent (672 of 1,560) of the population in a remote coastal Australian Aboriginal community aged 4.4 to 72.1 years participated in the study. RESULTS: Kidney size correlated closely with body size. Systolic blood pressure (SBP) was correlated inversely with kidney length and kidney volume, after adjusting for age, sex, and body surface area (BSA); a 1-cm increase in mean kidney length was associated with a 2.2-mm Hg decrease in SBP, and a 10-mL increase in mean kidney volume was associated with a 0.6-mm Hg decrease in SBP (P = 0.001). Mean kidney volume explained 10% of the variance in SBP in a multivariate model containing age, sex, and BSA. In addition to higher SBP, adults who had the lowest quartiles of kidney volume also had the highest levels of overt albuminuria (P = 0.044). CONCLUSION: Smaller kidneys predispose to higher blood pressures and albuminuria in this population. The lower volumes possibly represent kidneys with reduced nephron numbers, which might be related to an adverse intrauterine environment. Susceptibility to renal disease could be a direct consequence of reduced nephron numbers; the higher blood pressures with which they are associated could also contribute to, as well as derive from, this association. PMID- 14750092 TI - Treatment of pure membranous lupus nephropathy with prednisone and azathioprine: an open-label trial. AB - BACKGROUND: The aim of this study was to report the outcome of pure membranous lupus nephropathy treated with prednisone and azathioprine (AZA). METHODS: Consecutive patients with pure membranous lupus glomerulonephritis (World Health Organization [WHO] Va and Vb) from 4 regional hospitals were recruited for an open-label treatment trial consisting of prednisone and AZA. Remission status was evaluated at 12 months. Maintenance treatment with low-dose prednisone and AZA was continued indefinitely for those who achieved remission. Factors predictive of initial renal remission and subsequent relapse were studied by statistical analyses. RESULTS: Thirty-eight patients (31 women and 7 men) were studied. The mean age was 35.0 +/- 9.2 years, and the duration of systemic lupus erythematosus was 48.5 +/- 59 months. Seventeen (45%) patients had WHO class Va lupus nephritis, whereas 21 (55%) had class Vb disease. Two patients withdrew from the protocol because of idiosyncratic reactions to AZA. At 12 months, 24 (67%) patients achieved complete remission (CR), 8 (22%) achieved partial remission (PR), and 4 (11%) were treatment resistant. Patients who achieved CR or PR were maintained on low-dose prednisone and AZA. Over a mean follow-up period of 90.4 +/- 59 months, 6 (19%) patients had relapse of nephritis (proteinuric flare in 4 and nephritic flare in 2). The cumulative risk of renal relapse was 12% at 36 months and 16% at 60 months. No particular clinical variables were found to predict renal remission or relapses. Over a mean follow-up of 90 months, 13% of patients had decline of creatinine clearance by 20%, but none had doubling of serum creatinine. Renal outcome was not significantly worse in patients presenting with nephrotic syndrome. Treatment generally was well tolerated. CONCLUSION: A combination of prednisone and AZA is reasonably effective for the initial treatment of pure membranous lupus nephritis. Severe adverse effects are uncommon. The additional efficacy of AZA in comparison with prednisone alone has to be confirmed with randomized, controlled trials. PMID- 14750091 TI - Low-dose dual blockade of the renin-angiotensin system in patients with primary glomerulonephritis. AB - BACKGROUND: Treatment with agents interfering with the renin-angiotensin system retards the progressive course of proteinuric chronic renal disease. However, because of unwanted effects associated with such therapy, some patients cannot be treated with these drugs at all or may be administered only very small doses. To find an optimal nephroprotective strategy for these patients, we compared antiproteinuric effects of combination therapy with an angiotensin-converting enzyme inhibitor and angiotensin II type 1 receptor antagonist in very small doses with treatment with either agent alone at greater, but not maximal, doses. We compared the concomitant use of benazepril, 5 mg, and losartan, 25 mg, and monotherapy with these agents in doses 2-fold greater. METHODS: This is a randomized, open, crossover study of 3 treatments in 3 periods of 4 months each. Twenty-four patients with primary glomerulonephritis and nonnephrotic proteinuria, recognized previously as not able to be administered high doses of drugs from these classes, completed the protocol. RESULTS: Combined therapy decreased 24-hour proteinuria (-45.54% versus baseline) more effectively than either losartan (-28.17%; analysis of variance, P < 0.01) or benazepril (-20.19%; analysis of variance, P < 0.001) alone. Subgroup analysis showed that antiproteinuric effects of combination therapy, as well as losartan or benazepril alone, were significantly greater in patients with basal proteinuria greater than 2 g/24 h than in those with proteinuria less than this value (P < 0.001, P < 0.01, and P < 0.05, respectively). All therapies significantly decreased blood pressure (BP) compared with baseline, but there were no differences between treatments in BP changes. CONCLUSION: The study shows that combination therapy with very small doses of losartan and benazepril was more effective in reducing proteinuria than greater doses of either agent in monotherapy, and this greater antiproteinuric efficacy was independent of changes in BP. PMID- 14750093 TI - Outcomes and complications of pregnancy in women with primary hyperoxaluria. AB - BACKGROUND: Information about pregnancy in females with primary hyperoxaluria types I and II (PH-I, PH-II) is limited to isolated case reports. This study sought to determine the number and outcomes of pregnancies in a series of women with PH-I and PH-II, to assess the incidence of complications during pregnancy, and to characterize infant outcomes. METHODS: From a database of patients with PH followed at the Mayo Clinic, we identified 16 females who had been pregnant. A retrospective medical record review and telephone survey were performed. RESULTS: Forty pregnancies occurring between 1961 and 1998 were identified: 26 pregnancies in 11 patients with PH-I and 14 pregnancies in 5 patients with PH-II. Thirty (75%) of the pregnancies were carried to term, and 33 infants were born. Four miscarriages, 4 preterm births, and 2 elective abortions occurred. No maternal complications were reported in half of the pregnancies. In the remaining pregnancies, the most common complications were hypertension, urinary tract infection, and urolithiasis-associated symptoms. One PH-I patient developed pre eclampsia resulting in a stillborn infant, and another PH-1 patient developed hyperemesis gravidarum with a decline in renal function. Approximately 75% of the infants had no perinatal problems. All of the PH-I patients eventually required initiation of renal replacement therapy at a mean of 17.5 years following their first pregnancy. No PH-II patients have required renal replacement therapy. CONCLUSION: Overall, pregnancy appeared to be well tolerated in this series of women with primary hyperoxaluria, with 1 of 16 women experiencing loss of renal function during 1 of 40 pregnancies. PMID- 14750094 TI - Growth arrest specific protein 6/Axl signaling in human inflammatory renal diseases. AB - BACKGROUND: Growth arrest-specific gene 6 (Gas6) and its binding partner, the receptor tyrosine kinase Axl, are important mediators in experimental nephritis. The authors tested whether the Gas6/Axl signaling pathway participates in human renal diseases. METHODS: The authors compared 26 human renal specimens from patients with IgA nephritis, acute diffuse immune complex glomerulonephritis, acute lupus nephritis, antineutrophil cytoplasmic antibody--associated glomerulonephritis, acute transplant rejection, and normal renal tissue. Because reactive oxygen species are pivotal in inflammation, the authors tested whether the Axl/Gas6 expression is influenced by NADPH oxidase in vitro. RESULTS: Gas6 and Axl immunofluorescence was barely detectable in normal kidney. However, in disease Axl was copiously expressed in the small vessel media, glomeruli, distal tubules, and collecting ducts. Similarly, Gas6 was upregulated in the small vessel intima and media, all segments of the renal tubules, the brush border, and glomeruli. Gas6 and Axl upregulation was a prominent but nonspecific finding in these renal diseases. Cultured rat vascular smooth muscle cells and immortalized human mesangial cells were stimulated with angiotensin (Ang) II (1 x 10(-7) mol/L) for 6 or 18 hours. Confocal microscopy and Western blot showed Ang II dependent Gas6 and Axl expression. An antisense probe against the p22 phox unit of NADPH-oxidase suppressed Ang II-induced Gas6 and Axl expression. In addition, in p47 phox knockout cells Ang II-induced Gas6 and Axl expression were blocked. CONCLUSION: GAS6/Axl signaling is involved in human renal disease. The Ang II induced Gas6 and Axl expression may be dependent on NADPH-oxidase. Gas6 and Axl are important signaling molecules in human renal disease and may be potential therapeutic targets. PMID- 14750095 TI - Phenotypic heterogeneity in pediatric autosomal dominant polycystic kidney disease at first presentation: a single-center, 20-year review. AB - BACKGROUND: The presentation of autosomal dominant polycystic kidney disease (ADPKD) in childhood provides an insight into comorbidities and potential areas for interventions and investigation. METHODS: Phenotypic heterogeneity at the time of first presentation was studied with respect to age of diagnosis, mode of presentation, parental inheritance pattern, renal function, associated hypertension, and hyperlipidemia. Fifty-five children (median age of presentation, 8.7 years; 27% < 1 year) with ADPKD from 44 families followed up between March 1983 and March 2003 were reviewed. The diagnosis was based on family history and ultrasound confirmation of cysts. Progression of renal disease was followed over the study period (mean duration of follow-up, 4.9 years). RESULTS: A family history of ADPKD was known at presentation in 89%, which precipitated the screening diagnostic imaging in 59% of these children. Maternal inheritance was displayed in 51%, whereas 5% had no known family history of ADPKD. Bilateral renal findings were present in 78%. Hypertension (>95(th) percentile for age) was present in 22%, and hyperlipidemia was present in 54%. Renal function was not significantly diminished in 98% of patients with creatinine clearance > or =3rd percentile for age, and 7% had persistent proteinuria (>150 mg/d). No subjects had hepatic, splenic, or pancreatic cysts on ultrasound scan. A subpopulation of 10 patients had features of ADPKD dating back to prenatal ultrasound scans. All prenatal cases were characterized by bilateral renal findings, 90% had a known family history of ADPKD at the time of presentation, and 89% of these patients displayed maternal inheritance. Follow-up studies showed a persistence of hyperlipidemia despite pharmacotherapeutic treatment of hypertension, infrequent proteinuria, and sustained renal function in most patients. CONCLUSION: The results of this study show that many children at the time of first presentation have a significant prevalence of modifiable risk factors: hypertension, proteinuria, and hyperlipidemia, in the face of normal renal function. The results also show a unique presentation existing in prenatal subjects. PMID- 14750096 TI - Intrafamilial phenotype variability in patients with Gitelman syndrome having the same mutations in their thiazide-sensitive sodium/chloride cotransporter. AB - BACKGROUND: Gitelman syndrome (GS) most often results from mutations in the thiazide-sensitive sodium chloride cotransporter (NCC). Although the severity of symptoms may vary in patients who have the same mutations, a markedly different clinical presentation in family members with identical mutations is truly rare. METHODS: Five patients (3 women and 2 men) belonging to 2 unrelated Chinese families were investigated. All had chronic hypokalemia, renal potassium (K+) wasting, metabolic alkalosis, and normal blood pressure. Direct sequencing of both the NCC and CLCNKB genes were performed. RESULTS: The probands in each family were men. They had very severe hypokalemia and were symptomatic with episodes of paralysis. They had normal plasma magnesium concentrations, normal calcium excretion rates, and impaired maximal urine concentrating ability. In contrast, female family members were asymptomatic. They had laboratory findings typical of GS--less severe hypokalemia, hypomagnesemia, hypocalciuria, and intact maximal renal concentrating ability. Nevertheless, all patients had the same novel pair of NCC mutations and no mutations detected in CLCNKB. CONCLUSION: Differences in sex may help explain the different clinical presentations in these 2 Chinese families with novel NCC mutations. Hypomagnesemia and hypocalciuria are not always present in patients with GS. PMID- 14750097 TI - Coronary artery calcification measured with electron-beam computerized tomography correlates poorly with coronary artery angiography in dialysis patients. AB - BACKGROUND: Coronary artery calcification measured by electron-beam computerized tomography (EBCT) correlates with plaque burden and vessel stenosis and is predictive of future cardiac events in the general population. Uremic vascular calcification is common and widespread, tends to be medial as well as intimal, and may not relate solely to atherosclerotic lesions. Despite this difference and in the absence of any direct evidence in uremic patients, it is generally implied that coronary artery calcification equates to occlusive atherosclerosis. METHODS: We set out to compare the predictive value of coronary artery calcification assessed by EBCT with contemporaneous coronary angiography. We studied 18 patients with end-stage renal disease undergoing maintenance dialysis. Seventy two coronary vessels were analyzed for angiographic evidence of stenotic disease and correlated with individual vessel calcification score. RESULTS: There was no significant correlation between degree of vessel stenosis and calcification score for individual vessels in patients with a positive calcium scan. Specificity was 48% and the positive predictive value was 53%. However, a calcification score <20 strongly correlated with the absence of significant luminal narrowing, and a 0 calcification score had a negative predictive value of 87.5%. CONCLUSION: Coronary artery calcification measured by EBCT is not an accurate marker of the degree of vessel stenosis in coronary artery disease in uremic patients and should not be used as a single screening test for atherosclerotic coronary disease. PMID- 14750098 TI - Ability of non-high-density lipoprotein cholesterol and calculated intermediate density lipoprotein to identify nontraditional lipoprotein subclass risk factors in dialysis patients. AB - BACKGROUND: Non-high-density lipoprotein cholesterol (non-HDL-C) and calculated intermediate-density lipoprotein cholesterol (IDL-C) have been proposed as surrogate markers to estimate apolipoprotein B-containing lipoproteins. The purpose of this study was to determine the validity of non-HDL-C and calculated IDL-C to predict nontraditional lipoprotein risk factors among dialysis patients and to compare the prevalence of these nontraditional risk factors between dialysis modalities. METHODS: The authors performed a cross-sectional analysis comparing standard lipid profile with lipoprotein analysis via nuclear magnetic resonance (NMR) spectroscopy among 186 hemodialysis (HD) and peritoneal dialysis (PD) patients on modern lipid-lowering therapy. RESULTS: The PD group had a significantly higher low-density lipoprotein (LDL) particle concentration (P < 0.005), higher large very low-density lipoprotein (VLDL; P < 0.001), greater small dense LDL (P < 0.001), and lower protective large HDL (P < 0.005). Forty six (40%) of 118 subjects with LDL-C below goal had at least 1 nontraditional risk factor by NMR spectroscopy. The sensitivity of non-HDL-C method together with triglyceride (TG) value greater than 200 mg/dL (>2.26 mmol/L) to predict nontraditional risk was 13% and increased to 20% if TG values were excluded. A negative correlation was observed between LDL particle size and HDL-C (r2 = 0.269; P < 0.001); the sensitivity of HDL-C to predict LDL size was 92%. There was no relationship between measured IDL by NMR and calculated IDL-C (r2 = 0.005; P = 0.69). CONCLUSION: Non-HDL-C greater than 130 mg/dL (3.4 mmol/L) independent of TG values and HDL-C lower than 40 mg/dL (1.0 mmol/L) may predict nontraditional lipoprotein risk factors among dialysis patients. This is especially applicable to patients on PD, a modality associated with a more atherogenic lipoprotein profile. PMID- 14750099 TI - The metabolic response to ingested protein is normal in long-term hemodialysis patients. AB - BACKGROUND: Protein-energy malnutrition affects 30% to 50% of hemodialysis (HD) patients. This has been attributed to inadequate food intake, but may be caused by disturbances in utilization of ingested protein. METHODS: We studied protein kinetics during fasting and during ingestion of a protein-enriched meal to investigate possible metabolic differences between stable HD patients and control subjects. Whole-body protein kinetics was measured by means of a primed constant infusion of L[1-13C] valine. RESULTS: During fasting, whole-body protein balance was significantly less negative in HD patients compared with control subjects. During meal intake, protein balance was similar between HD patients and control subjects. Meal intake increased protein balance significantly in both groups, but not differently between the groups. Also, protein oxidation was decreased during fasting in HD patients compared with control subjects, but not during meal intake. CONCLUSION: We conclude that the rate of protein breakdown is lower in HD patients compared with control subjects, but the efficiency of protein utilization is normal in HD patients during a nondialysis day. PMID- 14750100 TI - Efficacy and cardiovascular tolerability of extended dialysis in critically ill patients: a randomized controlled study. AB - BACKGROUND: Extended dialysis is an increasingly used modality of renal replacement therapy that theoretically offers advantages of both intermittent and continuous therapies in the intensive care unit (ICU). METHODS: We randomly treated 39 ventilated critically ill patients with oliguric acute renal failure with either continuous venovenous hemofiltration (CVVH; n = 19; age, 50.1 +/- 3.2 years; Acute Physiology and Chronic Health Assessment II [APACHE II] score, 32.3 +/- 1.2; 79% sepsis) and a substitution fluid rate of at least 30 mL/kg/h for 24 hours or with extended dialysis for 12 hours (n = 20; age, 50.8 +/- 3.6 years; APACHE II score, 33.6 +/- 1.0; 85% sepsis). The latter was performed using an easy-to-handle, single-pass, batch dialysis system. All hemodynamic parameters were monitored invasively by means of an indwelling arterial catheter. RESULTS: Average mean arterial blood pressure, heart rate, cardiac output, systemic vascular resistance, and catecholamine dose were not significantly different in both therapies. Urea reduction rate was similar with extended dialysis compared with CVVH therapy (53% +/- 2% versus 52% +/- 3%; P = not significant) despite an average rate of substitution fluid with the latter of 3.2 +/- 0.1 L/h. This was corroborated by the finding of similar amounts of urea eliminated in the collected spent total hemofiltration and dialysis fluid. Correction of acidosis was accomplished faster with extended dialysis than CVVH, and the amount of heparin used was significantly lower with extended dialysis (P < 0.01). CONCLUSION: Extended dialysis combines excellent detoxification with cardiovascular tolerability, even in severely ill patients in the ICU. The technically simple dialysis system used offers flexibility of treatment time. PMID- 14750101 TI - Physicians' beliefs about racial differences in referral for renal transplantation. AB - BACKGROUND: Black patients with end-stage renal disease are much less likely than white patients to undergo renal transplantation, but previous research has shown that black patients are only slightly less likely to desire this procedure. A better understanding of physicians' views about racial differences in access to transplantation may help reduce disparities in care. METHODS: We surveyed 278 nephrologists in 4 US regions about quality of life and survival for black and white patients undergoing renal transplantation and reasons for racial differences in access to transplantation. We also surveyed 606 of their patients about their care. RESULTS: Physicians were less likely to believe transplantation improves survival for blacks than whites (69% versus 81%; P = 0.001), but similarly likely to believe it improves quality of life (84% versus 86%). Factors commonly cited by physicians as important reasons why blacks are less likely than whites to be evaluated for transplantation included patients' preferences (66%), availability of living donors (66%), failure to complete evaluations (53%), and comorbid illnesses (52%). Fewer physicians perceived patient-physician communication and trust (38%) or physician bias (12%) as important reasons. Black patients were less likely than white patients to report receiving some or a lot of information about transplantation (55% versus 74%; P = 0.006) when their physicians did not view patient-physician communication and trust as an important reason for racial differences in care. CONCLUSION: Nephrologists' views about the benefits of renal transplantation and reasons for racial differences in access to this procedure may affect how they present this treatment option to black and white patients. PMID- 14750102 TI - Clinical and histological presentation of 3 siblings with mutations in the NPHP4 gene. AB - Nephronophthisis (NPH) is an autosomal recessive kidney disease characterized by tubular basement membrane disruption, interstitial infiltration, and tubular cysts. NPH leads to end-stage renal failure in the first 2 decades of life. Four genes responsible for different types of NPH have been identified: NPHP1, NPHP2, NPHP3, and NPHP4. The NPHP1 gene encodes nephrocystin; NPHP2, inversin; NPHP3, nephrocystin-3; and NPHP4, nephrocystin-4. We report 3 siblings from a consanguineous family with NPH who were previously described as carrying a homozygous mutation in the NPHP4 gene. Renal imaging showed cysts in the children. The histological picture of NPHP4 showed the same characteristic features as those known for NPHP1 and NPHP3. Progression to end-stage renal disease occurred between the ages of 17 and 22 years. None of the renal transplants showed recurrence of the disease. Retinitis pigmentosa was absent in all affected family members. PMID- 14750103 TI - Unusual placement of a dialysis catheter: persistent left superior vena cava. AB - Many nephrologists perform clinical procedures, and perhaps the most common is placement of a noncuffed dialysis catheter to obtain vascular access necessary for immediate hemodialysis therapy. The right internal jugular vein frequently is the site of choice for placement of such catheters in most patients, but placement in the left internal jugular vein would not be unusual; for example, if another central catheter is present in the right internal jugular vein or there has been a failed attempt at that site. Nephrologists who place hemodialysis catheters in the left internal jugular vein should be aware of the existence of an anatomic variant, a persistent left superior vena cava, to prevent alarming misinterpretation and inappropriate clinical responses to routine chest radiographs taken to confirm adequate placement of such catheters. PMID- 14750104 TI - Pathologic classification of focal segmental glomerulosclerosis: a working proposal. PMID- 14750105 TI - Treatment of proliferative lupus nephritis. PMID- 14750106 TI - Disparities in transplantation: what should we do? PMID- 14750107 TI - An overlapping etiology of rapidly progressive glomerulonephritis. PMID- 14750109 TI - A critical appraisal for definition of hyperfiltration. PMID- 14750117 TI - A case of Langerhans'-cell histiocytosis with membranous nephropathy. AB - A 26-year-old man with a history of Langerhans'-cell histiocytosis (LCH) of the bone presented with nephrotic-range proteinuria. Renal biopsy results showed changes characteristic of membranous nephropathy. During the current hospitalization, the patient had 2 episodes of pulmonary embolism. LCH at this time was documented in the lymph nodes. The patient was treated with repeated courses of vinblastine and high doses of corticosteroids to achieve remission of the basic disease and the renal involvement. After 2 years, complete remission of both the lymphadenopathy and the nephrotic syndrome was achieved. The association of membranous nephropathy with LCH might be attributable to an underlying abnormality in the immune system, a paraneoplastic manifestation, or both. This is the first report of LCH associated with severe nephrotic syndrome caused by membranous nephropathy. PMID- 14750118 TI - Remission of collapsing focal segmental glomerulosclerosis following chemotherapy for myeloma. AB - Collapsing focal segmental glomerulosclerosis can be idiopathic but has been associated with human immunodeficiency virus infection as well as lymphoproliferative, myeloproliferative, and autoimmune disorders. It has a more rapid progression to end-stage renal failure. Treatment of this condition is poorly defined with no clear evidence of response to any treatment regimens. We report a case of collapsing focal segmental glomerulosclerosis in association with multiple myeloma that underwent partial remission following treatment for the underlying disease. PMID- 14750119 TI - Pulmonary embolism due to catheter fracture from a tunneled dialysis catheter. AB - The patient presented to the emergency room with hemoptysis and pleuritic chest pain. A chest x-ray revealed a broken dialysis catheter tip lodged in the pulmonary artery. The fractured catheter tip was removed via the femoral vein using a loop snare. As has been described for central lines and venous ports, a fractured catheter tip from a hemodialysis catheter may also lead to pulmonary embolism. PMID- 14750120 TI - Acute interstitial nephritis and pigmented tubulopathy in a patient after wasp stings. AB - Acute renal failure (ARF) is an unusual complication of wasp stings and mostly results from toxic-ischemic acute tubular necrosis. This patient, who was stung by a swarm of wasps, experienced an allergic reaction, rhabdomyolysis, intravascular hemolysis, and subsequent ARF. The originality of this case report is related to the finding of combined lesions of acute tubulointerstitial nephritis and acute tubular nephropathy. From our extensive literature review, it is the first case of a patient developing this type of injury after wasp stings, and the complications have been documented previously only in one form or another. Renal biopsy should be encouraged, especially for a patient with delayed recovery of renal function after wasp stings, to facilitate early steroid treatment for the patient with the histological change of acute interstitial nephritis. Early use of steroid therapy may hasten renal recovery by preventing the development of interstitial fibrosis. PMID- 14750121 TI - Esophageal varices as a rare complication of central venous dialysis tunneled cuffed catheter. AB - Long-term complications of tunneled cuffed catheters include infection and catheter and venous thrombosis. We present a case of a central venous dialysis tunneled cuffed catheter complicated with esophageal varices resulting from superior vena cava obstruction. PMID- 14750122 TI - Necrotizing glomerulonephritis caused by Bartonella henselae endocarditis. AB - Glomerulonephritis secondary to endocarditis is uncommon and usually associated with valvular infection by blood culture-positive bacteria. We report 3 cases of necrotizing glomerulonephritis associated with culture-negative endocarditis caused by Bartonella henselae. Two of the patients presented with renal abnormalities and were investigated for endocarditis after results of renal biopsy. All 3 patients had an immune complex-mediated necrotizing and crescentic glomerulonephritis with mesangial and capillary wall deposition of immunoglobulin M (IgM), IgG, and C3. Electron microscopy showed immune-type electron-dense deposits in the mesangium and segmental subendothelial (2 cases) or subepithelial (1 case) deposits. Patients were treated with antibiotics, including azithromycin or doxycycline and ceftriaxone or tobramycin. In addition, 2 patients were administered steroids and 2 patients underwent valve replacement surgery. The 2 patients who underwent cardiac surgery were discharged from the hospital with stable renal function. The third patient died 4 months after hospital admission of renal failure. In conclusion, glomerulonephritis caused by B henselae endocarditis is an immune complex-mediated disease characterized by segmental necrotizing and crescentic glomerular lesions that can respond to aggressive medical and surgical therapy. PMID- 14750123 TI - Contrast-induced translocational hyponatremia and hyperkalemia in advanced kidney disease. AB - BACKGROUND: In patients with advanced kidney disease, administration of effective osmolar agents may lead to hyponatremia, hyperkalemia, metabolic acidosis, and intravascular expansion. To our knowledge, osmolar effects of contrast agents in patients with advanced kidney disease have not been previously elucidated. METHODS: This retrospective case series includes 5 patients with advanced kidney disease who underwent diagnostic and/or therapeutic cardiac catheterization and developed hyponatremia. Blood chemistry tests were performed before and after the procedure. Hyponatremia is defined as a plasma sodium concentration less than 135 mEq/L (mmol/L). RESULTS: Mean precontrast and postcontrast sodium levels were 138.6 mEq/L (mmol/L) and 122.6 mEq/L (mmol/L), respectively. Plasma potassium levels increased after contrast exposure. There was no difference in degree of hyperkalemia between patients with and without diabetes. Plasma bicarbonate levels were noted to decrease after contrast exposure in 4 of 5 patients, with the exception of a patient undergoing continuous ambulatory peritoneal dialysis. There was a strong correlation between dose of contrast administered and change in sodium level, with a correlation coefficient of 0.91. CONCLUSION: These data suggest that large volumes of contrast may result in hypertonic hyponatremia through a dual effect of exogenous fluid dilution and translocation. The fluid translocated from intracellular to extracellular spaces may lead to a decrease in sodium, chloride, and bicarbonate levels. All patients developed hyperkalemia, which may be caused by solvent drag and/or passive diffusion, in addition to impaired excretory capacity. Patients with advanced kidney disease exposed to large volumes of contrast should be monitored for osmolar-induced chemical changes so that timely postcontrast dialytic therapy can be instituted. PMID- 14750125 TI - Quiz page. Severe metabolic acidosis, caused by an enteric augmentation of urinary tract and renal impairment. Decompensation was caused by lack of self catheterization. PMID- 14750126 TI - Flow cytometer electronics. AB - Flow cytometers, like all things electronic, have undergone rapid evolution since the introduction of the first commercial instruments 30 years ago. Cytometers then had so little automation and sophistication that the operator had to be expert in many different scientific disciplines to be able to coax any data at all out of the instrument. But trends in electronic components toward higher speed, smaller size, and lower power consumption have allowed the instruments to become faster, more sensitive, and actually quasi-intelligent. Automation of complex processes that once required considerable operator expertise, such as running a high-speed sort or correctly compensating a multicolor experiment, has expanded the number of flow cytometry applications and the number of flow cytometry users; there is no reason to expect that trend to stop. This review discusses the electronic components of a current "generic" flow cytometer from a functional point of view with the hope that a better understanding of what is going on "inside the box" will lead to a better understanding of how operator selectable instrument settings can influence the quality of the data produced. PMID- 14750127 TI - New method for the analysis of cell cycle-specific apoptosis. AB - BACKGROUND: In this study, a new method for the analysis of cell cycle specificity of apoptosis was designed by using a modified annexin V and propidium iodide (API) method. METHODS: Cells of the human promyelocytic HL-60 line treated with camptothecin (CPT) or ultraviolet light (UV) were labeled with fluorescein isothiocyanate-conjugated annexin V and prefixed with 1% methanol-free formaldehyde on ice, and their DNA was stained stoichiometrically with propidium iodide in the presence of digitonin. Cellular green and red fluorescences were measured by flow cytometry. RESULTS: Cell cycle specificity of apoptosis obtained by the API method and those analyzed for the presence of DNA strand breaks by using terminal deoxynucleotidyl transferase (TdT) assay were similar: CPT- or UV induced apoptosis preferentially in S- or G1-phase cells, respectively. When the internucleosomal DNA degradation was prevented by the serine protease inhibitor N tosyl-L-phenylalanine chloromethyl ketone, apoptotic cells could not be detected by the TdT assay but were identified by the API method. CONCLUSIONS: The API method, similar to the TdT assay, accurately detects the cell cycle phase specificity of apoptosis. Also, the API method appears to detect earlier stages of apoptosis than the TdT assay. PMID- 14750128 TI - Measurement of basal, substrate induced and total P-glycoprotein activity in bronchoalveolar lavage T-cell subsets. AB - BACKGROUND: P-glycoprotein (P-gp) is a member of the ABC transporter superfamily. P-gp activity can be detected by measuring efflux of fluorescent substrates such as rhodamine 123 (R123). Our objectives were to evaluate P-gp activity in T cells freshly isolated from bronchoalveolar lavage (BAL) and to develop a strategy to distinguish between basal, in vitro substrate-induced, and total P-gp activities. METHODS: Cells were obtained from blood (n = 44) and BAL (n = 34), stained for expression of CD3, CD4, CD8, and CD14, and incubated with R123 (0.13 microM) +/- cyclosporine (5 microM), a specific P-gp inhibitor. P-gp activity was detected as median fluorescence intensity (MFI) and percentage of cells falling below a pre established cutoff. RESULTS: BAL T cells displayed significant basal P-gp activity, which was most apparent when measured as percentage below the cutoff. Induced activity (difference between P-gp activity measured after load and efflux) was determined equally well when using the MFI or the percentage below cutoff parameter. Total activity was represented by the efflux parameters (MFI or percentage below cutoff) or by the activity-time area under the curve (AUC) method. The two efflux parameters correlated well but were insensitive to the time-dependent nature of dye efflux. In the AUC method, two samples with identical R123 brightness or percentage below cutoff after dye efflux can have very different total activities, depending on their basal activity. The AUC method was also most sensitive in distinguishing between P-gp activity in peripheral blood and resident lung T cells. Application of this methodology to the comparison of P-gp activity in BAL and peripheral CD8+ T cells best revealed the elevated total P-gp activity in BAL T cells. CONCLUSIONS: We have systematically evaluated several methodologies for analysis of P-gp activity and arrived at a novel and robust strategy amenable to standardization and evaluation of the effects of P-gp modulators in vivo and in vitro. PMID- 14750129 TI - Development of Herceptin resistance in breast cancer cells. AB - BACKGROUND: Herceptin, a humanized antibody to HER-2, is now utilized in the clinic for metastatic breast cancer treatment. The response rate for HER-2+ patients is only 30% and little is known as to mechanisms of resistance. The mechanism of Herceptin action is also unknown but has been related to cell cycle inhibition. METHODS: The effects of Herceptin and other antibody treatments were determined by cell counting and cell cycle analysis. HER-2 and p27 expression levels were analyzed by flow cytometry and levels of activated AKT were compared by Western blot analysis. Cellular HER-2 and p27 expression was measured by immunofluorescence. RESULTS: Herceptin treatment of BT-474 cells results in inhibition of cell growth and arrest in the G1 phase. The efficacy of growth arrest was not directly correlated to the binding affinity of antibodies to Her 2. Our laboratory has developed cell lines that are resistant to Herceptin treatment. In resistant cell lines, binding of antibodies is not hindered. However, Herceptin has completely lost the ability to inhibit cell proliferation. Yet, the mouse isotype 4D5 maintains significant inhibitory activity upon Herceptin-resistant clones. CONCLUSIONS: Herceptin binds effectively to Her-2 on the cell surface of Herceptin-resistant cell lines and the level of Her-2 expression on the cell surface is not downregulated. Herceptin resistance is not due to downregulation of levels of AKT protein expression, although, phosphorylation of AKT is enhanced in resistant lines and could have a role in resistance. Resistance appears to correlate with the loss of nuclear expression of the cyclin-dependent kinase inhibitor, p27, as defined by immunofluorescence and flow cytometry studies and cdk-2 binding studies. PMID- 14750130 TI - Flow cytometer for measurement of the light scattering of viral and other submicroscopic particles. AB - BACKGROUND: Light scattering is an essential parameter in flow cytometry, facilitating functions such as size measurement, discrimination of cell types on the basis of shape and morphology, detection of fluorescence-negative cells, and gating of fluorescence measurements. Light scattering measurement of viruses is generally not feasible with current flow cytometers due to their small size. The problem is aggravated by the fact that the light scattering of particles in this size range (< 200 nm) falls off with roughly the sixth power of their linear dimensions. METHODS: A new optical layout using darkfield illumination and detection has been developed. A 532-nm laser was used for excitation, and scattered light was collected with large aperture optics. RESULTS: Light scattering histograms of polymer particles with diameters of 70-300 nm were recorded without gating by other parameters. By extrapolation, a detection limit of about 50 nm was obtained. Different species of virus with sizes of approximately 100 nm also were recorded. CONCLUSIONS: Flow cytometric light scattering measurement of submicroscopic particles, in a size range that includes many viral species, is now feasible. The results indicate that it may be practically impossible to measure by flow cytometry the light scattering of particles smaller than 40 nm. PMID- 14750131 TI - Comparison of flow cytometry and laser scanning cytometry for the analysis of CD34+ hematopoietic stem cells. AB - BACKGROUND: Characterization of hematopoietic stem cells (HSCs) by laser scanning cytometry (LSC) was compared with conventional flow cytometry (FCM). The method was evaluated for application in the development of advanced cell culture substrates that were supposed to support the ex vivo expansion of HSC. For this purpose, adherent HSCs were grown in culture on thin polymer films coated with reconstituted collagen I fibrils and subsequently analyzed by LSC. METHODS: CD34+ HSCs were isolated from cord blood by immunomagnetic separation and cultivated on polymer films coated with reconstituted collagen I fibrils. Cell surface antigens (CD34, CD29) were stained with antibodies, and nuclei were labeled with a DNA stain (TO-PRO-3 iodide) that does not interfere with the fluorochromes of the antibodies. Fluorescence intensity of the adherent cells was measured by means of LSC. Before and after in vitro expansion for time periods of up to 7 days, suspension cells were analyzed with LSC and FCM. RESULTS: LSC-based analysis enabled reliable quantification of CD34+ cells with bright antigen expression before cell culture. At this stage, LSC and FCM data for CD34 expression at given HSC samples largely coincided. After in vitro expansion, LSC data deviated from FCM data for cells with dim CD34 antigen expression, whereas the fluorescence intensity of the CD29 antigen remained comparable. The deviation between LSC and FCM data for CD34dim was attributed to the better resolution of weak fluorescence by FCM. Based on the preceding evaluation of the method, LSC analysis could be applied to characterize HSCs cultivated on collagen I-coated polymer films without detachment of the cells from the substrate. CONCLUSIONS: LSC-based analysis allows for the automated evaluation of adherent HSCs. Although resolution of weakly expressed antigens can be achieved more precisely with FCM, the method provides a valuable tool to study interactions of HSCs with bioartificial substrates. PMID- 14750132 TI - Spatial pattern analysis of nitrergic neurons in the developing myenteric plexus of the human fetal intestine. AB - BACKGROUND: Enteric nervous system precursors derived from the neural crest migrate along defined pathways to colonize the bowel. The individual cells in different environments experience different growth, differentiation, and survival conditions. Hence, the spatial distribution of the neurons is determinant with regard to functional maturation. The question arises as to whether the distribution is random or nonrandom. METHODS: Nitrergic cells were visualized by means of nicotinamide adenine dinucleotide phosphate diaphorase histochemistry. Stained specimens were photographed, and the borders of the myenteric plexus and the nuclei of the nitrergic neurons were digitalized. Plexus Pattern Analysis software was used to count the nuclei of nitrergic neurons, calculate the proportions of the areas covered by the plexus and the gut wall, and perform randomization analyses. RESULTS: The distribution pattern of the nitrergic neurons changed markedly between weeks 14 and 22 of gestation. The nitrergic neurons were randomly distributed at week 14 but were aggregated in the plexus and within the individual ganglia at week 19. The dynamics of these changes exhibited regional differences. CONCLUSIONS: The results suggest that, in addition to the gut wall and the plexus, other intraganglionic constituents may contribute to the aggregation of nitrergic cells and such examinations should be extended to other cell types in the future. PMID- 14750133 TI - Simple, semiautomatic assay of cytostatic and cytotoxic effects of antitumor drugs by laser scanning cytometry: effects of the bis-intercalator WP631 on growth and cell cycle of T-24 cells. AB - BACKGROUND: Common assays of drug-induced cytotoxicity on adherent cells rely on cell trypsinization followed by count of live and dead cells. To estimate the cell cycle effects, cellular DNA content is analyzed by flow cytometry. This procedure is laborious and time consuming. The alternative viability assays, e.g., based on reduction of 3-(4,5-dimethylthiazol-2-yl) 2,5-diphenyl tetrazolium bromide, although rapid and convenient, do not provide information about individual cells or cell cycle effects and may be biased by growth imbalance. METHODS: The bladder carcinoma T-24 cells were seeded onto eight-chamber microscope slide-based tissue culture vessels. The novel antitumor drug, the bis intercalating anthracycline WP631, was administered at various concentrations to different chamber cultures on the same slide; the control cultures were left untreated. After 24, 48, and 72 h, the cultures were fixed, and cellular DNA was stained with 4,6-diamidino-2-phenyl indole (DAPI). The slides were scanned by laser scanning cytometry (LSC) to obtain the number of attached cells per culture chamber and reveal their cell cycle distribution. RESULTS: The cell growth and viability plots in the absence and presence of WP621 were constructed from the frequency of the attached cells per chamber. A 50% reduction in cell number was observed at the 75 nM concentration of WP321. Mitotic and postmitotic cells were identified based on high intensity of maximal pixel of DAPI fluorescence. An increase in proportion of cells in G2 was seen at 75-300 nM of WP631. Relatively few (<12%) apoptotic cells, identified by the presence of DNA strand breaks, remained attached in the WP631-treated cultures. CONCLUSIONS: Because late apoptotic cells detach during culturing, the cells that remain attached in the multi-chamber cultures represent predominantly live cells; the deficit in their number compared with the untreated cultures, recorded by LSC during scanning, provides information about the degree of cytostatic and cytotoxic effects of the studied drug. The possibility to demonstrate the cell cycle distribution, including a distinction between G2 and M cells, provides an additional advantage of this assay. Other parameters that may be associated with the cell cycle perturbation or with induction of apoptosis also can be measured in the same cultures by using the multiparameter capabilities of LSC. Each measured cell can be relocated for imaging or measurement after subsequent staining with other probes. PMID- 14750134 TI - Quantitation of MHC tetramer-positive cells from whole blood: evaluation of a single-platform, six-parameter flow cytometric method. AB - BACKGROUND: Quantitation of antigen-specific T cells provides an insight into the development and dynamics of T-cell responses in tumor immunology and infectious diseases. Soluble major histocompatibility class I tetramers are widely used to monitor immune responses; however, variations due to handling and analysis are likely to confound comparisons between different experiments and laboratories. METHODS: Whole blood from healthy donors was stained with HLA-A*0201/tetramers specific for an epitope of phosphoprotein 65, the immunodominant antigen in cytomegalovirus infection. With the help of Trucount tubes, a single-platform, four-color flow cytometric assay was established to obtain absolute counts of tetramer-positive cells. Various staining and gating strategies were evaluated. RESULTS: The no-wash method was a quick and straightforward procedure for the quantitation of tetramer-positive events from whole blood. The level for background staining was low. This information about the intra-assay-related variation and the physiologic variation will allow validation and interpretation of data in future studies. CONCLUSIONS: The method is highly reliable and can be standardized for multiple experiments. It is therefore suitable for the direct ex vivo analysis of antigen-specific T cells in a variety of clinical settings such as infectious, autoimmune, or neoplastic diseases and can be implemented as a tool for multicenter studies. PMID- 14750135 TI - Experience with focused abdominal sonography for trauma (FAST) in 313 pediatric patients. AB - PURPOSE: The use of focused abdominal sonography for trauma (FAST), which detects free fluid in the abdomen and pelvis, for the assessment of blunt abdominal trauma is gaining acceptance worldwide and has been described extensively in the general medical literature. The precise application of this technique in pediatric patients, however, has yet to be established. The aim of this study was to assess the utility of FAST in pediatric trauma patients by comparing the results of this technique with those of CT and explorative laparotomy (ELAP). METHODS: We retrospectively reviewed the medical records and sonographic examinations of pediatric patients who had sustained multiple traumatic injuries for which they were treated at our hospital during a 20-month period. For all patients, FAST had been the initial screening examination for blunt abdominal trauma. We compared the FAST findings, which had been recorded as positive or negative, with the findings on CT or ELAP, which were considered definitive. RESULTS: A total of 313 patients (204 boys and 109 girls) with a mean age of 7.1 years were included in the study. The FAST finding had been negative in 274 patients, of whom 201 had had no clinical signs of abdominal injury and had been managed conservatively without complications. CT had been performed in 109 patients and ELAP in 11. FAST had yielded 3 false-negative and 2 false-positive results. The sensitivity, specificity, and accuracy of FAST were 92.5%, 97.2%, and 95.5%, respectively. CONCLUSIONS: FAST is an effective tool in screening pediatric trauma patients for blunt abdominal trauma. PMID- 14750136 TI - Color Doppler sonographic signs of respiration-dependent hepatofugal portal flow. AB - PURPOSE: The role of respiration in modulating blood flow in the portal vein is unclear. The aim of this study was to investigate the phenomenon of respiration dependent periodic hepatofugal portal venous blood flow as detected on color Doppler sonography. METHODS: Within 1 year, we identified 13 patients with respiration-dependent reversal of blood flow in the portal vein that was diagnosed on color Doppler sonography. This phenomenon was investigated by color Doppler sonographic examination of the portal venous flow during both mid inspiration breath-holding and a respiratory cycle including deep inspiration; evaluation of hepatic vein Doppler waveforms as normal (triphasic) or decreased (flattened); and echocardiographic examination to determine the presence or absence of tricuspid regurgitation. RESULTS: The patients' median age was 53 years (range, 26-87 years). Seven of the 13 patients had heart disease (tricuspid regurgitation) with or without liver disease, 3 had liver disease without heart disease, and 3 had other diseases with no evidence of heart or liver disease. On Doppler sonography, 10 of the 13 patients had increased portal venous pulsatility (7 of the 10 had tricuspid regurgitation; the other 3 did not); the remaining 3 patients had neither increased pulsatility nor tricuspid regurgitation. Sonographic follow-up within 4 weeks in 4 of the 13 patients revealed loss of the respiration-dependent hepatofugal portal flow. CONCLUSIONS: Respiration-dependent hepatofugal portal flow is a rare finding associated with periodic portal hypertension in patients with right heart insufficiency and liver disease. Its clinical significance is unclear. Among our patients, its occurrence was predominantly associated with an increased venous pulsatility index due to tricuspid regurgitation or venous outflow obstruction. Further study is needed to investigate whether periodic respiration-dependent hepatofugal portal flow is predictive of the occurrence of continuous flow reversal. PMID- 14750137 TI - Characteristic imaging features of breast fibroadenomas in women given cyclosporin A after renal transplantation. AB - PURPOSE: This retrospective study was conducted to determine the characteristic imaging and histopathologic features of breast fibroadenomas in kidney-transplant recipients given chronic cyclosporin A immunosuppressive therapy after the transplantation and to compare these characteristics with those from a control group. METHODS: From January 1, 1990, through December 31, 1999, 486 women underwent renal transplantation at our institution. All patients subsequently received immunosuppressive therapy with cyclosporin A and prednisolone. Ten (2%) of these women had developed breast fibroadenomas during this chemotherapy. We compared the data obtained on this group's fibroadenomas with those obtained from those in the control group, which comprised 100 women with fibroadenomas but who had never undergone organ transplantation or immunosuppressive therapy. RESULTS: Twenty-two fibroadenomas developed in the 10 transplant recipients. Eight of those 10 had multiple lesions, and 7 were affected bilaterally. The mean diameter of the fibroadenomas was 4.2 +/- 2.5 cm. Mammographically, the lesions were round or oval high-density masses with well-circumscribed margins and no calcification or spiculation. Sonographically, the lesions were relatively highly echogenic and had a lower ratio of the longitudinal to the anteroposterior diameter (L /AP) than did those in the control group. Histopathologically, the features of these lesions were generally typical of fibroadenomas, but some were more typical of malignant lesions. Among the 100 control patients, 146 fibroadenomas developed; 33 women had multiple lesions, and 12 were affected bilaterally. Their fibroadenomas had a mean diameter of 2.1 +/- 1.5 cm, and the imaging features of these lesions were typical of fibroadenomas. CONCLUSIONS: The fibroadenomas that developed in kidney-transplant recipients given chronic cyclosporin A and prednisolone immunosuppressive therapy had a tendency to be multiple, bilateral, and larger than those that developed in the control patients. These fibroadenomas also exhibited some imaging features that differed from those of typical fibroadenomas that develop in women who have not undergone organ transplantation or immunosuppressive therapy. PMID- 14750138 TI - Characteristic sonographic findings of Warthin's tumor in the parotid gland. AB - PURPOSE: The aim of this study was to define the characteristic sonographic features of Warthin's tumors in the parotid gland, thus enhancing the ability to make a preoperative diagnosis of this disease process. METHODS: We retrospectively evaluated the sonograms of Warthin's tumors of the parotid gland that had been confirmed by histopathologic examination of surgically excised specimens from patients treated at our institution over a 2-year period. We recorded the echogenicity, shape, margin, and internal structure of the tumors and categorized them into 2 groups on the basis of size (< 5 cm versus > or =5 cm in the largest diameter). RESULTS: Eighteen patients (16 men and 2 women) with a mean age of 57 years (range, 29-82 years) were included in the analysis. One patient had 2 tumors (1 in each parotid gland); the other 17 patients each had only 1 tumor. Our review of the sonographic findings revealed that all 19 tumors were hypoechoic compared with the normal parenchyma of the affected parotid glands. Most of the tumors were ovoid, had well-defined margins, and contained multiple irregular, small, sponge-like anechoic areas. Tumors that were 5 cm or greater in diameter had a higher proportion of cystic content than smaller lesions had and in some cases were composed almost entirely of cystic material. CONCLUSIONS: Our evidence suggests that the sonographic appearance of a Warthin's tumor in the parotid gland is distinctive: a well-defined round or ovoid hypoechoic mass containing microcystic anechoic areas. These sonographic features constitute useful criteria in the preoperative diagnosis of Warthin's tumor in the parotid gland, although sonographically guided fine-needle aspiration biopsy may be needed to confirm the diagnosis. PMID- 14750139 TI - Simple new technique for sonographic measurement of the thyroid in neonates and small children. AB - PURPOSE: The aim of this study was to determine whether the ratio of the thyroid width (Th) to the trachea width (Tr) is a useful technique for sonographic estimation of thyroid size in neonates and small children. METHODS: We prospectively performed sonographic examination of the thyroid gland in 30 pediatric subjects ranging in age from 1 week to 12 years. The sum of the maximum width of the left thyroid lobe and the maximum width of the right lobe was considered the Th. Each subject's thyroid was classified as small, normal sized, or large on the basis of the Th value in reference to the subject's body height. The ratios of the Th to the Tr were then calculated and compared among the 3 groups by 1-way analysis of variance. Correlation and regression analyses were performed to determine the correlation between body height and Tr. A p value of less than 0.05 was considered significant. RESULTS: The mean Th/Tr ratios (+/- standard deviations) for the 3 groups were as follows: small, 1.25 +/- 0.19; normal sized, 2.09 +/- 0.19; and large, 4.10 +/- 2.42. The difference in the Th/Tr ratios between the 3 groups was statistically significant (p < 0.0001). A significant positive correlation was found between Tr and body height (p < 0.001; r = 0.85). CONCLUSIONS: The Th/Tr ratio is a simple, practical parameter for estimating the size of the thyroid gland in neonates and small children. PMID- 14750140 TI - Counterclockwise barber-pole sign on prenatal three-dimensional power Doppler sonography in a case of duodenal obstruction without intestinal malrotation. AB - Three-dimensional (3D) power Doppler sonographic imaging provides a 3D view of the blood vessels. This technique reportedly has advantages over other forms of sonography in visualizing normal and abnormal fetal vascular anatomy. We report the case of a 36-year-old pregnant woman in whom 3D power Doppler sonography with simultaneous gray-scale imaging was performed at 32 weeks' gestational age to investigate an intestinal obstruction in the fetus that was suspected on the basis of 2-dimensional sonographic findings. The 3D sonograms revealed that the superior mesenteric artery and vein of the fetus were wound counterclockwise, forming the "barber-pole" sign. At 38 weeks' menstrual age, the female infant was delivered vaginally. Only duodenal atresia and annular pancreas, but not intestinal malrotation, were found postnatally. Surgical reconstruction of the duodenum was performed, with no major complications. Previous research suggests that the counterclockwise barber-pole sign is a normal finding. On the basis of our case, we believe that this "negative finding" might be used to exclude the diagnosis of midgut volvulus, a condition that would be expected to cause these vessels to wrap around in a clockwise direction. PMID- 14750142 TI - Subserous uterine adenomyosis mimicking an adnexal mass on sonography. AB - Uterine adenomyosis usually manifests as diffuse disease involving the myometrium and the endometrial-myometrial junction, but it may also manifest as a focal lesion. It is usually only a few millimeters in diameter but may sometimes be larger. We report the case of a 32-year-old woman with a large isolated mass in the uterine wall. Transvaginal sonography demonstrated the cystic nature of the mass and its characteristic hemorrhagic pattern, whereas CT confirmed its uterine origin. The patient underwent laparoscopic resection of the mass, and pathologic examinations led to the diagnosis of adenomyosis. PMID- 14750141 TI - Migration of an intrauterine contraceptive device to the ovary. AB - We present the case of a 37-year-old woman with a history of 2 consecutive insertions of intrauterine contraceptive devices (IUDs) 3 years before she was referred to us for sonographic evaluation of lower abdominal pain. The first of the IUDs was presumed to have been expelled spontaneously, and 3 months after insertion of the second device, the patient had begun experiencing lower abdominal pain. Medical treatment with antibiotics and spasmolytics had been unsuccessful. We performed transvaginal sonography, which revealed the presence of an IUD in the uterus and a 2-cm linear metallic echogenic area in the left ovary, believed to represent another IUD. Anteroposterior radiography confirmed that there were 2 IUDs in the pelvis, and CT demonstrated 1 IUD in the uterus and another in the left ovary. The patient underwent laparoscopic removal of the ovarian IUD and was discharged in good condition. To our knowledge, this is the first report of migration of an IUD to the ovary detected on transvaginal sonography. We recommend consideration of this possibility during evaluation of women with unexplained chronic pelvic pain. PMID- 14750143 TI - Sonographic diagnosis of placental teratoma. AB - Placental teratomas are very rare, and the features that distinguish them on sonography and allow their differentiation from other placental tumors have not been fully described. Prenatal recognition of this tumor is prognostically useful because, unlike other neoplasms, placental teratoma is benign and almost never associated with congenital deformities in the fetus. We describe the case of a 27 year-old pregnant woman in whom prenatal color Doppler sonographic examination performed during early-stage labor revealed a heterogeneous mass at the placental margin. This lesion, which measured 10 x 8 x 5 cm, had an echogenic focus consistent with calcification and hyperechoic foci consistent with fat. Placental teratoma and fetus acardius amorphus were considered in the differential diagnosis, but the segmental organization and umbilical cord-like structures that characterize the latter diagnosis were absent. The sonographic diagnosis, placental teratoma, was confirmed postnatally by histopathologic examination. The neonate, a healthy boy, was delivered vaginally at term. Although the presence of tissues of varied echogenicity, such as calcification, fat, and fluid, and the absence of both polarity and an umbilical cord-like structure support the sonographic diagnosis of placental teratoma, fetus acardius amorphus should be considered in the differential diagnosis. Histopathologic examination may be needed to establish a definitive diagnosis in such cases. PMID- 14750144 TI - Asymmetric parasitic dicephalus conjoined twins. AB - In this report, we describe the case of a 19-year-old primigravida pregnant woman who had been hospitalized for 2 days with bleeding and a diagnosis of imminent abortion. Sonographic examination revealed a single fetus of approximately 17 weeks' gestation that initially appeared normal except for a mass on its neck. Further evaluation of the mass identified it as an anencephalic fetal head that was joined to the body with deformed cervical vertebrae. We diagnosed asymmetric parasitic dicephalus conjoined twins. The parents elected to terminate the pregnancy. Labor was induced, and the patient delivered a 20-cm, 208-g male fetus. The fetus had 4 normal limbs and a normal abdomen. Both its heads had a cleft lip, and its spine was U shaped with closed spina bifida at the spinal union in the thoracolumbar area. The patterns of conjoined twins are infinitely varied, but they can be generally classified according to standardized terms based on the anatomic site of their union. Cases of conjoined twins occur so rarely that it is important to learn as much as possible from each case. PMID- 14750145 TI - Odorant receptor expression in the mouse cerebral cortex. AB - Mammalian odorant receptors have been known to be involved not only in odorant detection but also in neuronal development of olfactory sensory neurons. We have examined a possibility of odorant receptor expression in nonolfactory neurons in the mouse. Mouse odorant receptors (M71, C6, and OR3), two of which were already shown to be functionally activated by odorants in heterologous systems, were detected by polymerase chain reactions (PCRs) from the cerebral cortex but not from other brain tissues. Degenerate PCR further suggested that other odorant receptors were also expressed in the mouse cerebral cortex. One of these receptors showed high sequence-match with a putative chick odorant receptor OR7 transiently expressed in the notochord during development. In situ hybridization detected signals for M71 and C6 receptors in the layer II cortical pyramidal neurons located in the occipital pole. In the M71-IRES-tauLacZ mouse, in which M71 expression was genetically marked with tauLacZ, X-gal staining signals were mostly localized in the layer II neurons in the occipital pole, being consistent with the in situ hybridization result. Fluorescent immunohistochemistry using anti-beta-galactosidase antibody further detected the tauLacZ signals in the same cells. X-gal staining began at P3, peaked at P8, and continued to adults, although signals gradually decreased. These data showed that at least a few odorant receptors are expressed not only in olfactory sensory neurons but also in pyramidal neurons in the cerebral cortex, possibly playing an important role either in chemical detection of exogenous or endogenous ligands or in a developmental process such as axon guidance and target recognition. PMID- 14750146 TI - Presynaptic protein kinase C controls maturation and branch dynamics of developing retinotectal arbors: possible role in activity-driven sharpening. AB - Visual activity refines developing retinotectal maps and shapes individual retinal arbors via an NMDA receptor-dependent mechanism. As retinal axons grow into tectum, they slow markedly and emit many transient side branches behind the tip, assuming a "bottlebrush" morphology. Some branches are stabilized and branch further, giving rise to a compact arbor. The dynamic rate of branch addition and deletion is increased twofold when MK801 is used to block NMDA receptors, as if this prevents release of a stabilizing signal such as arachidonic acid (AA) from the postsynaptic neuron. In optic tract, AA mediates NCAM and L1 stimulation of axon growth by activating presynaptic protein kinase C (PKC) to phosphorylate GAP 43 and stabilize F-actin, and, if present in tectum, this growth control pathway could be modulated by postsynaptic activation. To test for the effects on arbor morphology of blocking PKC or AA release, we examined DiO-labeled retinal axons of larval zebrafish with time-lapse videomicroscopy. Bath application of the selective PKC inhibitor bisindolylmaleimide from 2 or 3 days onward doubled the rate at which side branches were added and deleted, as seen with MK801, and also prevented maturation of the arbor so that it retained a "bottlebrush" morphology. In order to selectively block the PKC being transported to retinal terminals, we injected the irreversible inhibitor calphostin C into the eye from which the ganglion cells were labeled, and this produced both effects seen with bath application. In contrast, there were no effects of control injections, which included Ringers into the same eye and the same dose into the opposite eye (actually much closer to the tectum of interest), to rule out the possibility that the inhibitor leaked from the eye to act on tectal cells. For comparison, we examined arbors treated with the NMDA blocker MK801 at half-hour time-lapse intervals, and detected the twofold rise in rates of branch addition and deletion previously reported in Xenopus larvae, but not the structural effect seen with the PKC inhibitors. In addition, we could produce both effects seen with PKC inhibitors by using RHC80267 to block AA release from DAG lipase, indicating that AA is the main drive for PKC activation. Thus, the results show a distinct role of AA and presynaptic PKC in both maturation of arbor structure and in the dynamic control of branching. The effects on branch dynamics were present regardless of the level of maturity of arbor structure. The fact that they mimicked those of MK801 suggests that presynaptic PKC may be involved in the NMDA receptor-driven stabilization of developing retinal arbors. PMID- 14750147 TI - Changes in retinal expression of neurotrophins and neurotrophin receptors induced by ocular hypertension. AB - Open angle glaucoma is defined as a progressive and time-dependent death of retinal ganglion cells concomitant with high intraocular pressure, leading to loss of visual field. Because neurotrophins are a family of growth factors that support neuronal survival, we hypothesized that quantitative and qualitative changes in neurotrophins or their receptors may take place early in ocular hypertension, preceding extensive cell death and clinical features of glaucoma. We present molecular, biochemical, and phenotypic evidence that significant neurotrophic changes occur in retina, which correlate temporally with retinal ganglion cell death. After 7 days of ocular hypertension there is a transient up regulation of retinal NGF, while its receptor TrkA is up-regulated in a sustained fashion in retinal neurons. After 28 days of ocular hypertension there is sustained up-regulation of retinal BDNF, but its receptor TrkB remains unchanged. Throughout, NT-3 levels remain unchanged but there is an early and sustained increase of its receptor TrkC in Muller cells but not in retinal ganglion cells. These newly synthesized glial TrkC receptors are truncated, kinase-dead isoforms. Expression of retinal p75 also increases late at day 28. Asymmetric up-regulation of neurotrophins and neurotrophin receptors may preclude efficient neurotrophic rescue of RGCs from apoptosis. A possible rationale for therapeutic intervention with Trk receptor agonists and p75 receptor antagonists is proposed. PMID- 14750148 TI - Inducible transcript expressed by reactive epithelial cells at sites of olfactory sensory neuron proliferation. AB - The continuous replacement of cells in the spiny lobster olfactory organ depends on proliferation of new cells at a specific site, the proximal proliferation zone (PPZ). Using representational difference analysis of cDNA, we identified transcripts enriched in the PPZ compared to the mature zone (MZ) of the organ. The 12 clones identified included four novel sequences, three exoskeletal proteins, a serine protease, two protease inhibitors, a putative growth factor, and a sequence named PET-15 that has similarity to antimicrobial proteins of the crustin type. PET-15 mRNA was only detected in epithelial cells. It was abundant in all epithelial cells of the PPZ, but was only detected in the MZ at sites of damage to the olfactory organ. PET-15 mRNA was increased by types of damage that are known to induce proliferation of new olfactory sensory neurons in the olfactory organ. It increased in the PPZ after partial ablation of the olfactory organ and in the MZ after shaving of aesthetasc sensilla. These ipsilateral effects were mirrored by smaller increases in the undamaged contralateral olfactory organ. These contralateral effects are most parsimoniously explained by the action of a diffusible signal. Because epithelial cells are the source of proliferating progenitors in the olfactory organ, the same diffusible signal may stimulate increases in both cellular proliferation and PET-15 mRNA. The uniformity of expression of PET-15 in the PPZ epithelium suggests that the epithelial cells that give rise to new olfactory sensory neurons are a subset of cells that express PET-15. PMID- 14750149 TI - Odor-evoked responses in the olfactory center neurons in the terrestrial slug. AB - The procerebrum (PC) of the terrestrial mollusk Limax is a highly developed second-order olfactory center consisting of two electrophysiologically distinct populations of neurons: nonbursting (NB) and bursting (B). NB neurons are by far the more numerous of the two cell types. They receive direct synaptic inputs from afferent fibers from the tentacle ganglion, the primary olfactory center, and also receive periodic inhibitory postsynaptic potentials (IPSPs) from B neurons. Odor-evoked activity in the NB neurons was examined using perforated patch recordings. Stimulation of the superior tentacle with odorants resulted in inhibitory responses in 45% of NB neurons, while 11% of NB neurons showed an excitatory response. The specific response was reproducible in each neuron to the same odorant, suggesting the possibility that activity of NB neurons may encode odor identity. Analysis of the cycle-averaged membrane potential of NB neurons revealed a correlation between the firing rate and the membrane potential at the plateau phase between IPSPs. Also, the firing rate of NB neurons was affected by the frequency of the IPSPs. These results indicate the existence of two distinct mechanisms for the regulation of NB neuron activity. PMID- 14750150 TI - Identification of a proctolin preprohormone gene (Proct) of Drosophila melanogaster: expression and predicted prohormone processing. AB - Proctolin was the first insect neuropeptide to be sequenced and has been the subject of many physiological and pharmacological studies in insects and crustaceans. We have identified a Drosophila gene (CG7105, Proct) encoding a precursor protein containing the neuropeptide proctolin (RYLPT). In situ hybridization with a riboprobe to the Proct gene revealed a distribution of transcript in neurons of the larval central nervous system (CNS) matching that seen with antiserum to proctolin. An antiserum raised to a sequence in the precursor downstream of proctolin labeled the same neurons as those seen with the antiproctolin antisera. The predicted protein encoded by Proct has a single copy of the RYLPT sequence that directly follows the predicted signal peptidase cleavage point and precedes a consensus recognition site for a furinlike processing endoprotease. Ectopic expression of Proct in the CNS and midgut via the GAL4-UAS system, using an Actin5C-GAL4 driver, confirmed that the predicted preproproctolin can be processed to generate immunoreactive proctolin peptide. Pupae over-expressing Proct displayed a 14% increase in heart rate, providing evidence in support of a cardioacceleratory endocrine function for proctolin in Drosophila. The distribution of proctolin suggests roles as a neuromodulator in motoneurons and interneurons, and as a neurohormone that could be released from brain neurosecretory cells with terminations in the ring gland. PMID- 14750151 TI - TBX2/TBX3 transcriptional factor homologue controls olfactory adaptation in Caenorhabditis elegans. AB - Although transcriptional factors are known to play important roles in synaptic plasticity, their role in olfactory adaptation has not been studied well. Here we report that Ce-TBX-2, the TBX2/TBX3 transcriptional factor homologue of the nematode Caenorhabditis elegans, is involved in olfactory adaptation. Two missense hypomorphic mutations in this gene confer abnormality in adaptation, but not chemotaxis, to all the odorants sensed by AWC olfactory neurons. The Ce-tbx-2 gene is expressed in AWB, AWC, ASJ, and many pharyngeal neurons, but expression in AWC neurons is sufficient for normal adaptation. Unexpectedly, the protein product is localized mostly in cytoplasm. The AWC neurons in the mutants retain their characteristic morphology and many marker gene expressions, suggesting that the mutants are abnormal in neural functions rather than neuronal differentiation. The results of this study imply that some of the mammalian T-box family proteins, which play central roles in embryonic development, may also control functions like neural plasticity in differentiated neurons. PMID- 14750152 TI - Regulation of dendritic length and branching by semaphorin 3A. AB - The molecular mechanisms that control dendritic development are still unclear. Semaphorin 3A, a class III semaphorin, has been shown to regulate the radial orientation of pyramidal neurons in the developing neocortex. Here, we investigate the effects of Sema3A on the development of dendritic topology. Neocortical slices from Sema3A null mutant mice were cultured and neurons were transfected with GFP, reconstructed, and compared with neurons from wild-type and heterozygote littermates. We also added exogenous Sema3A to cultured wild-type neocortical slices to further test its effects on dendritic development. We document reductions in dendritic length and branching in Sema3A null mice and increases in dendritic length and branching after the addition of exogenous Sema3A to wild-type neurons. We conclude that Sema3A is necessary for the elaboration of second and third order dendritic branches in pyramidal neurons. PMID- 14750153 TI - Estrogen contributes to seasonal plasticity of the adult avian song control system. AB - Songbirds show dramatic neural plasticity as adults, including large-scale anatomical changes in discrete brain regions ("song control nuclei") controlling the production of singing behavior. The volumes of several song control nuclei are much larger in the breeding season than in the nonbreeding season, and these seasonal neural changes are regulated by plasma testosterone (T) levels. In many cases, the effects of T on the central nervous system are mediated by neural conversion to estradiol (E(2)) by the enzyme aromatase. The forebrain of male songbirds expresses very high levels of aromatase, in some cases adjacent to song control nuclei. We examined the effects of aromatase inhibition and estrogen treatment on song nuclei size using wild male songbirds in both the breeding and nonbreeding seasons. In breeding males, aromatase inhibition caused the volume of a telencephalic song control nucleus (HVC) to decrease, and this effect was partially rescued by concurrent estrogen replacement. In nonbreeding males, estradiol treatment caused HVC to grow to maximal spring size within 2 weeks. Overall, these data suggest that aromatization of T is an important mediator of song control system plasticity, and that estradiol has neurotrophic effects in adult male songbirds. This study demonstrates that estrogen can affect adult neural plasticity on a gross anatomical scale and is the first examination of estrogen effects on the brain of a wild animal. PMID- 14750155 TI - Functional and anatomical organization of floccular zones: a preserved feature in vertebrates. PMID- 14750156 TI - Functional compartmentalization in the flocculus and the ventral dentate and dorsal group y nuclei: an analysis of single olivocerebellar axonal morphology. AB - The cerebellar cortex consists of multiple longitudinal bands defined by their olivocerebellar projection. Single olivocerebellar axons project to a narrow longitudinal band in the cerebellar cortex and to the cerebellar nucleus with their axon collaterals. This olivocortical and olivonuclear organization is related to the functional compartmentalization of the cerebellar system. To reveal the detailed morphologic organization in the flocculus and the cerebellar and vestibular nuclei, we examined olivocerebellar projection by reconstructing the entire trajectories of 19 single olivofloccular axons and by anterograde mapping with biotinylated dextran in the rat. The flocculus was composed of 12 longitudinal band-shaped compartments that subdivided 5 previously described zones. These longitudinal bands were innervated differentially by the caudal and rostral portions of the dorsal cap (DC) and the ventrolateral outgrowth (VLO) and the rostral pole of the medial accessory olive. Single olivofloccular axons with an average of 5.1 climbing fibers usually projected to a single longitudinal band in the flocculus and to the ventral dentate or dorsal group y nucleus with their collaterals. DC neurons projected moderately to the rostrolateral portion of the ventral dentate nucleus, whereas VLO neurons projected densely to the medial portion of the ventral dentate nucleus and the dorsal group y nucleus with rostrocaudal topography. DC and VLO neurons did not project to the vestibular nuclei, although floccular Purkinje cells projected to the vestibular, ventral dentate, and dorsal group y nuclei. The fine morphologically identified longitudinal bands and topographic olivonuclear projections were correlated with previous electrophysiologically defined functional zones in the flocculus and inferior olive. PMID- 14750157 TI - Primary afferent terminal sprouting after a cervical dorsal rootlet section in the macaque monkey. AB - We examined the role of primary afferent neurons in the somatosensory cortical "reactivation" that occurs after a localized cervical dorsal root lesion (Darian Smith and Brown [2000] Nat. Neurosci. 3:476-481). After section of the dorsal rootlets that enervate the macaque's thumb and index finger (segments C6-C8), the cortical representation of these digits was initially silenced but then re emerged for these same digits over 2-4 postlesion months. Cortical reactivation was accompanied by the emergence of physiologically detectable input from these same digits within dorsal rootlets bordering the lesion site. We investigated whether central axonal sprouting of primary afferents spared by the rhizotomy could mediate this cortical reactivation. The cortical representation of the hand was mapped electrophysiologically 15-25 weeks after the dorsal rootlet section to define this reactivation. Cholera toxin subunit B conjugated to horseradish peroxidase was then injected into the thumb and index finger pads bilaterally to label the central terminals of any neurons that innervated these digits. Primary afferent terminal proliferation was assessed in the spinal dorsal horn and cuneate nucleus at 7 days and 15-25 postlesion weeks. Labeled terminal bouton distributions were reconstructed and the "lesion" and control sides compared within each monkey. Distributions were significantly larger on the side of the lesion in the dorsal horn and cuneate nucleus at 15-25 weeks after the dorsal rootlet section, than those mapped only 7 days postlesion. Our results provide direct evidence for localized sprouting of spared (uninjured) primary afferent terminals in the dorsal horn and cuneate nucleus after a restricted dorsal root injury. PMID- 14750158 TI - Development of GABA-immunoreactive cells in the spinal cord of the sea lamprey, P. marinus. AB - The lamprey spinal cord increases in length and size during all its life cycle; thus, it is expected that new cells will be generated. This expectation suggests that the locomotor circuits must be continuously remodeled. Key elements in the cellular network controlling locomotor behavior are inhibitory cells. Here, we studied the gamma-aminobutyric acid-immunoreactive (GABA-ir) cells in the lamprey spinal cord during postembryonic development. Three major populations of GABA-ir cells were identified according to their distribution: those located in the gray matter, those contacting the cerebrospinal liquid (LC cells), and those located in the white matter. The results show (1). the number of GABA-ir cells per segment increase from prolarvae (<10 mm) to adulthood; (2). the lower number of GABA-ir cells in 100 microm of spinal cord is 66 +/- 7, found in premetamorphic larvae, and the highest is 107 +/- 6, found in postmetamorphic animals; (3). the gray matter and LC GABA-ir cells show different variations in number depending on the developmental period. Thus, in the 10-mm larvae, the gray matter GABA-ir cells are more abundant than LC cells, whereas in the young postmetamorphic specimens, the contrary occurs. Most of the GABA-ir cells located in the white matter were classified as edge cells. They increase in number from the beginning of the prolarval period, where there are not white matter-positive cells, to the middle larval period, where there are 9 +/- 4 GABA-ir edge cells per segment. This value was unaltered in later periods, where GABA-ir edge cells represent 20 30% of the total number of edge cells per segment. The increase in number of GABA ir cells in these populations during a specific point of the lamprey life cycle may indicate different inhibitory requirements of the locomotor circuit at different developmental periods. PMID- 14750159 TI - Electron microscopy of degenerative changes in the chick basilar papilla after gentamicin exposure. AB - We present a sequential study of the substructural alterations in the chick basilar papilla at the earliest signs of hair cell degeneration. Three-day posthatch chicks received a single injection of gentamicin (300 mg/kg) and were killed at 6, 8, 12, 15, 18, 21, and 24 hours after the injection. The basilar papillae were studied by conventional transmission electron microscopy. Examination was limited to the basal region, where all hair cells are eliminated by this treatment. As early as 8 hours and clearly by 12 hours, altered fine structure was seen in hair cells. Changes included rounding and swelling of the hair cells, condensation of nuclear chromatin, dissolution of ribosomes, dilatation of the mitochondria, and accumulation of inclusion bodies and lysosomes. By 15-18 hours, lysosomes increased and became denser, afferent terminals appeared swollen, and the first cell extrusion was seen. Efferents were unaffected, and supporting cells, though having inclusion bodies now, retained normal intercellular junctions. By 21-24 hours, large regions of complete hair cell loss were composed of expanded supporting cell processes with normal appearing intercellular junctions and portions of extruded hair cells, partially attached to the supporting cell surface. These observations demonstrate that auditory hair cells undergo a rapid and controlled process of hair cell extrusion that allows preservation of the reticular lamina and minimal contamination of surrounding structures by intracytoplasmic contents of the damaged hair cells. PMID- 14750160 TI - Organization of parabrachial projections from the spinal trigeminal nucleus oralis: an anterograde tracing study in the rat. AB - In recent years, we have accumulated data showing that the spinal trigeminal nucleus oralis (Sp5O) contributes to the processing of somatosensory inputs from the orofacial region. Although the parabrachial area (PB) represents the main brainstem relay for autonomic, nociceptive, and gustatory afferents, few data are available regarding the topographical distribution of the efferent projections from the Sp5O to the PB. We have addressed this question with the rat, by using the anterograde tracer Phaseolus vulgaris leucoagglutinin. A dense trigeminoparabrachial pathway from the Sp5O toward, predominantly, the ipsilateral PB was revealed. Projections come mainly from the dorsal part of the Sp5O that was found to innervate densely the medial, external medial, and ventral lateral subnuclei. In contrast, the ventral part of the Sp5O projected almost exclusively to an as yet not formally described region, located dorsally and laterally to the lateral tip of the brachium conjunctivum, close to the Kolliker Fuse nucleus. These results suggest that distinct regions within the Sp5O may be involved in the processing of gustatory and nociceptive information. PMID- 14750161 TI - Immunohistochemistry and neural connectivity of the Ov shell in the songbird and their evolutionary implications. AB - The neuropeptide immunohistochemistry and neural connectivity of areas surrounding the thalamic auditory nucleus (the nucleus ovoidalis [Ov]), as well as the areas to which it is connected, were investigated in a songbird, the Bengalese finch. The results showed that met-enkephalin was present in the Ov shell and most of the areas connected to it, but not in the Ov core. Anterograde and retrograde tracing studies showed that the Ov shell was more widely connected than the Ov core. The Ov shell was mainly connected to: 1). areas flanking the primary telencephalic auditory field (i.e., fields L2b, L1, and L3) and areas surrounding the robust nucleus of the archistriatum (RA); 2). several hypothalamic areas such as the nucleus ventromedialis hypothalami (VMN) and the nucleus anterior medialis hypothalami (AM). Some of these areas connected to the Ov shell are thought to be involved in auditory mediated neurosecretory activities. These results, which are similar to those reported previously in non songbirds, suggest that the Ov shell and other surrounding areas of auditory and song-control nuclei are conserved in birds. These findings are discussed in terms of the evolution of the core-and-surround organization of auditory and song control nuclei. PMID- 14750162 TI - Components of the reelin signaling pathway are expressed in the spinal cord. AB - The Reelin signaling pathway in the brain involves the binding of Reelin to very low-density lipoprotein receptors (VLDLR) and apolipoprotein E receptor 2 (ApoER2). After Reelin binds the lipoprotein receptors on migrating neurons, the intracellular adaptor protein Disabled-1 (Dab1) becomes phosphorylated, ultimately resulting in the proper positioning of cortical neurons. Previous work showed that Reelin also affects the positioning of sympathetic preganglionic neurons (SPN) in the spinal cord (Yip et al. [2000] Proc Natl Acad Sci USA 97:8612-8616). We asked in the present study whether components of the Reelin signaling pathway in the brain also function to control SPN migration in developing spinal cord. Results showed that Reelin and reelin mRNA are found adjacent to migrating SPN. In addition, dab1 mRNA and protein are expressed by migrating SPN, and dab1-null mice show abnormal SPN migration similar to that seen in reeler. Finally, vldlr and apoER2 are also expressed in migrating SPN, and mice lacking both vldlr and apoER2 show aberrant SPN location that is identical to that of reeler and dab1-null mice. Because molecules known to be involved in Reelin signaling in the brain are present in the developing spinal cord, it is likely that the Reelin signaling pathways in the brain and spinal cord function similarly. The relative simplicity of the organization of the spinal cord makes it a potentially useful model system with which to study the molecular and cellular function of the Reelin signaling pathway in control of neuronal migration. PMID- 14750163 TI - Mechanisms of genomic instability in human cancer: insights from studies with human papillomavirus oncoproteins. AB - Genomic instability is a hallmark of most human cancers including high-risk human papillomavirus (HPV)-associated anogenital neoplasia. The two HPV-encoded oncoproteins, E6 and E7, can independently induce chromosomal abnormalities. We summarize the current state of knowledge concerning HPV-induced genomic instability and discuss its significance in the context of human carcinogenesis. PMID- 14750164 TI - CDH1 and CDH13 methylation in serum is an independent prognostic marker in cervical cancer patients. AB - Cervical cancer is the principal cause of death due to cancer in women. Five-year survival rate ranges from 15-80%, depending on the extent of the disease. New predictive markers for relapse may increase survival rates by improving treatment of patients at high risk for relapse. The gene products of CDH1 and CDH13, namely E-cadherin and H-cadherin, play a key role in cell-cell adhesion. Inactivation of the cadherin-mediated cell adhesion system, caused by aberrant methylation, is a common finding in human cancers. To test the hypothesis that CDH1/CDH13 methylation is a prognostic marker in cervical cancer we determined the methylation status of CDH1/CDH13 in serum samples from 93 cervical cancer patients. Methylation analysis was carried out using MethyLight. Aberrant methylation of the 5'-region of CDH1 or CDH13 was observed in 43% (40 of 93) of the patients. Cervical cancer patients with unmethylated CDH1/CDH13 in serum samples showed significantly better disease-free survival in univariate and multivariate analysis. Median disease-free survival for CDH1/CDH13 methylation negative and positive patients was 4.3 years and 1.2 years, respectively. Our results suggest that detection of aberrant methylation of CDH1/CDH13 may be of potential use as a marker for selecting cervical cancer patients at high risk for relapse who could benefit from additional systemic therapy. PMID- 14750165 TI - Resveratrol modulates the phosphoinositide 3-kinase pathway through an estrogen receptor alpha-dependent mechanism: relevance in cell proliferation. AB - Resveratrol (RES), a natural phytoalexin, has antiproliferative activity in human derived cancer cells and in rodent models of tumor development. We have previously shown that RES induced apoptotic death in estrogen-responsive MCF-7 human breast cancer cells. Recent data have indicated that the estrogen receptor alpha (ERalpha), through interaction with p85, regulates phosphoinositide 3 kinase (PI3K) activity, revealing a physiologic, nonnuclear function of the ERalpha potentially relevant in cell proliferation and apoptosis. In our study, using MCF-7, we have analyzed the ability of RES to modulate the ERalpha dependent PI3K pathway. Immunoprecipitation and kinase activity assays showed that RES increased the ERalpha-associated PI3K activity with a maximum stimulatory effect at concentrations close to 10 microM; concentrations >50 microM decreased PI3K activity. Stimulation of PI3K activity by RES was ERalpha dependent since it could be blocked by the antiestrogen ICI 182,780. RES did not affect p85 protein expression but induced the proteasome-dependent degradation of the ERalpha. Nevertheless, the amount of PI3K immunoprecipitated by the ERalpha remained unchanged in presence of RES, indicating that ERalpha availability was not limiting PI3K activity. Phosphoprotein kinase B (pPKB/AKT) followed the pattern of PI3K activity, whereas RES did not affect total PKB/AKT expression. PKB/AKT downstream target glycogen synthase kinase 3 (GSK3) also showed a phosphorylation pattern that followed PI3K activity. We propose a mechanism through which RES could inhibit survival and proliferation of estrogen-responsive cells by interfering with an ERalpha-associated PI3K pathway, following a process that could be independent of the nuclear functions of the ERalpha. PMID- 14750166 TI - Dynamic and intracellular trafficking of P-glycoprotein-EGFP fusion protein: Implications in multidrug resistance in cancer. AB - In our present study, a P-glycoprotein-EGFP (P-gp-EGFP) fusion plasmid was constructed and functionally expressed in HeLa cells to investigate the intracellular localization and trafficking of P-glycoprotein (P-gp). Using immunocytochemistry and fluorescent confocal microscopy techniques, colocalization studies showed that after transfection, P-gp-EGFP was progressively transported from the endoplasmic reticulum (ER) to the Golgi and finally to the plasma membrane within 12-48 hr. The degree of intracellular accumulation of daunorubicin was related to the particular localization of P-gp EGFP. Significant daunorubicin accumulation occurred in transfected cells when P gp-EGFP was localized predominantly within the ER, and accumulation remained high when P-gp-EGFP was mainly localized in the Golgi. However, there was little or no intracellular accumulation of daunorubicin when P-gp-EGFP was localized predominantly on the plasma membrane. Blocking the intracellular trafficking of P gp-EGFP with brefeldin A (BFA) and monensin resulted in inhibition of traffic of P-gp-EGFP and retention of P-gp-EGFP intracellularly. Intracellular accumulation of daunorubicin also increased in the presence of BFA or monensin. Our study shows that P-gp-EGFP can be used to define the dynamics of P-gp traffic in a transient expression system, and demonstrates that localization of P-gp on the plasma membrane is associated with the highest level of resistance to daunorubicin accumulation in cells. Modulation of intracellular localization of P gp with agents designed to selectively modify its traffic may provide a new strategy for overcoming multidrug resistance in cancer cells. PMID- 14750167 TI - Resistance of pancreatic cancer to gemcitabine treatment is dependent on mitochondria-mediated apoptosis. AB - Palliative chemotherapy with gemcitabine, a common mode of treatment of pancreatic cancer, has little influence on patients' survival. We investigated the impact of anti-apoptotic Bcl-xL protein and its antagonist Bax on gemcitabine induced apoptosis in human pancreatic carcinoma cells in vitro and in vivo. The level of Bcl-xL and Bax expression was determined in 3 established pancreatic cancer cell lines that differ in their sensitivity to gemcitabine-mediated apoptosis. Bcl-xL and Bax genes were transduced into Colo357 cells by retroviral infection. In addition, cells were transfected with c-FLIP to assess involvement of CD95 and caspase-8. The impact of Bax/Bcl-xL expression on gemcitabine sensitivity in vivo was evaluated in orthotopic Colo357 tumors in SCID mice. The apoptotic index revealed a strong inverse correlation between Bcl-xL expression and gemcitabine-induced apoptosis in the pancreatic carcinoma cell lines tested. Caspase-8 and Bid were cleaved in Colo357 cells exposed to gemcitabine, and there was no correlation with either Bcl-xL or with Bax expression. In contrast, the lack of mitochondrial transmembrane potential transition, release of cytochrome-c and absence of caspase-9- and PARP-cleavage showed a strong correlation with Bcl xL expression. Expression of c-FLIP significantly increased the resistance towards gemcitabine. Orthotopically growing Colo357-bcl-xl tumors in SCID mice were refractory to gemcitabine treatment, and in contrast to the in vitro data, Colo357-bax tumors exhibited a 12-fold greater tumor regression than Colo357-wild type tumors in the control group. Gemcitabine-induced apoptosis involves the mitochondria-mediated signaling pathway. A functional restoration of this pathway appears to be essential to overcome the resistance mechanisms of pancreatic tumor cells and to improve the response to therapy as demonstrated by Bax overexpression in a clinically relevant tumor model. PMID- 14750168 TI - Retinoblastoma protein in human renal cell carcinoma in relation to alterations in G1/S regulatory proteins. AB - The retinoblastoma gene product (pRb) is the main substrate for cyclin-dependent kinases (CDKs) during the G1/S transition. Aberrations in cell cycle regulatory proteins, which have been observed in many malignancies, can theoretically cause increased phosphorylation of pRb due to unbalanced CDK activities. The expression and phosphorylation of pRb and potential associations to cell cycle aberrations in renal cell carcinomas (RCC) has only partly been clarified. We therefore evaluated the presence of pRb and the level of pRb-phosphorylation in 216 RCCs arranged in tissue microarrays by using different pRb-antibodies, including pRb phosphospecific antibodies. Most RCCs (95%) expressed pRb, while cases with the low pRb levels, potentially indicative for pRb-inactivation, were few. In order to detect secondary alterations to a potential pRb-inactivation, the p16 expression was also monitored. None of the tumors exhibited increased p16 levels, confirming that pRb-inactivation is rare in RCC. Phosphorylated pRb was detected in approximately 50% of the RCCs, using Western blotting or immunohistochemistry. The immunohistochemical ppRb(ser807/811) levels were associated with high proliferation, cyclin D1, cyclin E and p27 protein content. Surprisingly, there was no association between pRb-phosphorylation and clinicopathological data. In summary, pRb seemed to be functional and aberrations in G1/S-regulatory proteins were associated with increased phosphorylation of pRb and proliferation. The data supports that pRb might be one of the main cell cycle regulators in RCC. PMID- 14750169 TI - Promoter hypermethylation of E-cadherin and its abnormal expression in Epstein Barr virus-associated gastric carcinoma. AB - Promoter hypermethylation of various tumor-related genes is extremely frequent in Epstein-Barr virus (EBV)-associated gastric carcinoma (EBVaGC). To investigate the significance of the promoter methylation in EBVaGC, we focused on one of the important proteins in the carcinogenesis of the stomach, E-cadherin. Methylation specific PCR analysis (MSP) was applied to surgically resected gastric carcinomas, together with immunohistochemistry, PCR-based analysis of mutations and allelic loss, and site-specific MSP of E-cadherin gene. By MSP, nearly all of the carcinomas showed aberrant methylation of E-cadherin promoter in EBVaGC (21/22), and the frequency of this aberration was significantly higher than that in EBV-negative gastric carcinoma (GC; 45/81; p = 0.0003). According to immunohistochemistry of E-cadherin, the frequency of abnormal staining pattern in EBVaGC (87%) was comparable to that in the diffuse type (80%), but higher than that in the intestinal type of EBV-negative GC (47%). Promoter methylation was well correlated with abnormal staining pattern in EBVaGC, but not in EBV-negative GC. Neither mutation nor allelic loss of E-cadherin was observed in EBVaGC. Methylation status of E-cadherin within each carcinoma was heterogeneous as far as examined. Thus, in addition to the known association involving p16, we determined that promoter methylation-mediated silencing of E-cadherin gene was also closely associated with the development of EBVaGC, although it becomes heterogeneous within a given tumor along its progression. PMID- 14750170 TI - Apc10.1: an ApcMin/+ intestinal cell line with retention of heterozygosity. AB - APC10.1 is a new intestinal cell line derived from ApcMin/+ mice that retains both the heterozygous Apc genotype and a nonactivated Wnt signaling pathway and displays an early neoplastic phenotype. Although tumorigenic both in immunodepressed and in immunocompetent syngeneic mice, it requires a high cell dose and a long latency. Its epithelial/intestinal origin is shown, in a gene expression profile, by the expression of epithelial transcripts (such as cytokeratin and laminin isoforms) and of developmental regulatory genes (such as Tcf-4, Hnf3beta, p21, Ihh, Hes1) necessary for, or involved in, the maintenance of intestinal stem cells. The lack of activation of the Wnt cascade in APC10.1 cells is shown both by the expression profile of Wnt target genes and by the standard TCF reporter assay. APC10.1 cell line is a novel in vitro model that can contribute to a better understanding of the clinical evolution of familial adenomatous polyposis and to finding of new prophylactic and therapeutic approaches. Supplementary material for this article can be found on the International Journal of Cancer website at http://www.interscience.wiley.com/jpages/0020-7136/suppmat/index.html. PMID- 14750171 TI - Acetylation of histones associated with the p21WAF1/CIP1 gene by butyrate is not sufficient for p21WAF1/CIP1 gene transcription in human colorectal adenocarcinoma cells. AB - Butyric acid is well recognized as a histone deacetylase (HDAC) inhibitor, and changes in histone acetylation are thought to alter gene expression. The mechanism by which sodium butyrate (NaB) induces p21WAF1/CIP1, a critical gene involved in the antiproliferative effect of NaB, was studied at the chromatin level. Using chromatin immunoprecipitation (ChIP) assay, acetylation of histone H3 was observed at the proximal region of the promoter within 30 min of NaB exposure and this extended to the distal region within 2 hr. By contrast, histone H4 was acetylated both at the proximal and the distal regions of the promoter within 30 min. NaB did not influence other histone modifications. NaB stimulated recruitment of the transcription factors ZBP89 and Sp1 as well as GCN5, but did not influence recruitment of Sp3, HDAC1, p300, or CBP. As recruitment of HDAC1 to the promoter appeared not to account for NaB-induced changes in histone acetylation, we aimed to influence HDAC activity by altering its phosphorylation status. The kinase inhibitor, H7, suppressed p21WAF1/CIP1 mRNA in both the absence and the presence of NaB without influencing the butyrate-induced hyperacetylation of H3 and H4 associated with the p21WAF1/CIP1 promoter. These results suggest that acetylation of histones at the p21WAF1/CIP1 promoter is not sufficient for NaB to exert antiproliferative effects via transcription of the p21WAF1/CIP1 gene. Induction of p21WAF1/CIP1 transcription by the phosphatase inhibitor, okadaic acid, in the absence of changes in association of acetylated histones with the p21WAF1/CIP1 promoter provides further evidence of the importance of phosphorylation to p21WAF1/CIP1 transcription. PMID- 14750172 TI - Endothelial cells co-cultured with wild-type and dominant/negative p53 transfected glioblastoma cells exhibit differential sensitivity to radiation induced apoptosis. AB - We performed expressional profiling of isogenic glioblastoma cell lines U87-Lux8 and U87-175.4. These cell lines differ in that U87-Lux8 expresses wild-type p53 and U87-175.4 expresses a dominant-negative p53 (175(His) mutation). DNA array analysis and real-time PCR measurements demonstrated that basal expression and response to irradiation were different in these isogenic glioblastoma cell lines. These differences included genes involved in growth regulation and genes associated with cell-to-cell and cell/ECM communications. Co-cultivation of U87 175.4 and U87-Lux8 with HUVE cells demonstrated that U87-175.4 cells suppress the angiogenic phenotype of HUVEC and increase their sensitivity to radiation-induced apoptosis compared to co-culture of U87-Lux8/HUVEC. These data suggest that blockade of p53 function may alter the communication between tumor cells and endothelial cells such that endothelial cells exhibit an increase in radiosensitivity. These findings may have important implications for the treatment of glioblastoma tumors and other human cancers. PMID- 14750173 TI - Identification of biomarkers for the initiation of apoptosis in human preneoplastic colonocytes by proteome analysis. AB - Development of resistance of cells toward proapoptotic signals is regarded as one of the key processes that allow tumor development. To identify proteins that are crucial for the initiation of apoptosis in NCOL-1 human preneoplastic colonocytes, we analyzed the proteome of cells exposed to the flavonoids flavone and quercetin that differ in their ability to induce apoptosis although they possess similar structures. Both flavonoids inhibited proliferation and induced differentiation of NCOL-1 cells but only quercetin committed the cells to apoptosis. The accessible proteome of NCOL-1 cells was separated by 2D polyacrylamide-gelelectrophoresis and proteins with changed expression level were identified by peptide mass fingerprints of tryptic digests of the protein spots. A pre-fractionation of soluble and lipophilic proteins was used to enhance the resolution of analysis. After exposure to the test compounds for 24 hr, 73 proteins displayed changed steady state levels in case of quercetin and 32 in case of flavone. Several heat-shock proteins, annexins and cytoskeletal caspase substrates were regulated by quercetin but not by flavone and these protein classes are known to play a role in apoptosis induction and execution. Whereas proteins like lamin A, C and desmoplakin, are indicators that apoptosis has already proceeded, others, such as annexin IV or protein kinase C-beta play a pivotal role in the early phases of apoptosis. In conclusion, proteome analysis allowed the identification of marker proteins that are involved in the initiation of apoptotic cell death in preneoplastic colonocytes and those may help to develop new strategies for cancer prevention. PMID- 14750174 TI - Genetic and epigenetic alterations of the blood group ABO gene in oral squamous cell carcinoma. AB - Loss of histo-blood group A and B antigen expression is a frequent event in oral carcinomas and is associated with decreased activity of glycosyltransferases encoded by the ABO gene. We examined 30 oral squamous cell carcinomas for expression of A and B antigens and glycosyltransferases. We also examined DNA from these tumors for loss of heterozygosity (LOH) at markers surrounding the ABO locus at chromosome 9q34, for loss of specific ABO alleles, and for hypermethylation of the ABO promoters. Loss of A or B antigen expression was found in 21 of 25 tumors (84%) and was a consistent feature of tumors lacking expression of A/B glycosyltransferases. LOH at 9q34 was found in 7 of 27 cases (26%), and one case showed microsatellite instability. Among 20 AO/BO cases, 3 showed loss of the A/B allele and 3 showed loss of the O allele. Analysis of the proximal ABO promoter by methylation-specific PCR and melting curve analysis showed hypermethylation in 10 of 30 tumors (33.3%), which was associated with loss of A/B antigen expression. ABO promoter hypermethylation was also found in hyperplastic or dysplastic tissues adjacent to the tumors, suggesting that it is an early event in tumorigenesis. Collectively, we have identified molecular events that may account for loss of A/B antigen expression in 67% of oral squamous cell carcinomas. PMID- 14750175 TI - Single nucleotide polymorphisms result in impaired membrane localization and reduced atpase activity in multidrug transporter ABCG2. AB - ABCG2/MXR/ABCP1/BCRP is a member of the ATP-binding cassette membrane transporter, which consists of six transmembrane regions and one ATP-binding cassette. The transporter is known to be involved in the efflux of various anticancer compounds such as mitoxantrone, doxorubicin and topoisomerase I inhibitor. In this study, we analyzed the effects of polymorphisms in ABCG2, V12M and Q141K on transporter function. When polarized LLC-PK1 cells were transfected with variant ABCG2, drug-resistance to topoisomerase I inhibitor of cells expressing V12M or Q141K was less than 1/10 that of wild-type ABCG2 transfected cells, and was accompanied by increased drug accumulation and decreased drug efflux in the variant ABCG2-expressing cells. We further elucidated the molecular mechanisms of the transport dysfunction by investigating membrane localization and ATPase activity. Confocal microscopic analysis revealed that apical plasma membrane localization of V12M was disturbed, while the localization of wild-type transporters occurred specifically in the apical plasma membrane of polarized LLC PK1 cells. Also, ATPase activities measured in the membrane of SF9 cells infected with variant ABCG2 showed that Q141K decreased activity by 1.3 below that of wild type ABCG2. In addition, kinetic analysis of ATPase activity showed that the K(m) value in Q141K was 1.4-fold higher than that of wild-type ABCG2. These results indicated that naturally occurring SNPs alter transport functions of ABCG2 transporter and analysis of SNPs in ABCG2 may hold great importance in understanding the response/metabolism of chemotherapy compounds that act as substrates for ABCG2. PMID- 14750176 TI - Genetic alterations in the peritumoral stromal cells of malignant and borderline epithelial ovarian tumors as indicated by allelic imbalance on chromosome 3p. AB - Stromal accumulation of hyaluronan in epithelial ovarian cancers is an independent predictor of tumor spreading and unfavorable outcome of the disease. We started to screen for chromosomal causes of this accumulation by studying deletions in 3p21.3, a region harboring 3 hyaluronidase genes (HYAL1-3) among other potentially important tumor suppressors. Using 6 microsatellite markers from this region, allelic imbalance was found in 60-87% of the informative tumor cells microdissected from histologic sections of 58 patients with epithelial ovarian cancer. However, adjacent stromal cells originally intended as controls showed allelic imbalance at a frequency almost as high as the tumor cells (52 80%). A further laser capture microdissection on 10 borderline tumors also showed a high rate of allelic imbalance, both in the epithelial and stromal cells, but with a pattern slightly different from cancers. Allelic imbalance in the tumor epithelium or stroma was not correlated with the accumulation of hyaluronan or clinicopathologic parameters, including tumor stage and grade. The results suggest that factors other than inactivation of the HYAL1-3 genes are responsible for hyaluronan accumulation in epithelial ovarian tumors. Moreover, the results indicate that the stromal cells of the epithelial ovarian cancers not only respond to the signals from malignant epithelium but also have themselves undergone genetic alterations in markers partly identical to those in the cancer epithelial cells and may actively contribute to the development of the tumor from its early stages to the late determinants of patient mortality. PMID- 14750177 TI - Esophageal squamous cell cancer in patients with head and neck cancer: Prevalence of human papillomavirus DNA sequences. AB - An etiologic role for human papillomavirus (HPV) infections in either head and neck (HNC) or esophageal carcinogenesis remains debatable. Patients with head and neck cancer are at high risk for developing a second esophageal squamous cell cancer (ESCC). The aim of our study was to determine whether HPV infections play a role in this multifocal carcinogenesis. Samples from 2 groups of HNC patients were studied: Random esophageal biopsies were collected from the first group of 60 patients who had been screened for asymptomatic ESCC. The second group consisted of 21 patients with pairs of HNC and ESCC. Both the fresh frozen biopsy samples of the first group and the paraffin-embedded specimens of the second group were evaluated for the presence of HPV DNA sequences by PCR amplification, cloning and sequencing. HPV DNA sequences were detected in 66.7% of normal/inflammatory (34/51) and dysplastic and malignant (6/9) esophageal tissues from HNC patients being screened endoscopically. Similarly, in the second group of 21 patients with both HNC and ESCC, HPV DNA sequences were demonstrated in 13 (61.9%) of the HNC biopsies and in 14 (66.7%) of the ESCC biopsies. The prevalence of high-risk-type HPV 16 was low (5/51, 9.8%) in normal/inflammatory esophageal mucosa but higher (10/24, 47.6%) in ESCC. The low-risk HPV 11 was present in 37.3% (19/51) of normal/inflammatory, 66.7% (4/6) of dysplastic and 28.9% (13/45) of the carcinoma samples. The same HPV type was present in only 3/21 pairs of HNC and ESCC samples, suggesting that a clonal expansion from the HNC to a subsequent ESCC, or visa versa, is unlikely. The high prevalence of "low risk" HPV infections points to the need for studies on possible interactions of these infections with the use of alcohol and tobacco in the pathogenesis of these tumors. PMID- 14750178 TI - CD4+ T cell-mediated HER-2/neu-specific tumor rejection in the absence of B cells. AB - HER-2/neu (HER-2) is a cell surface proto-oncogene that is often overexpressed in carcinomas. Passive administration of anti-HER-2 antibodies in breast cancer patients has achieved promising results, but less is known about the role of antibodies in active immunization. We asked whether B cells/antibodies are needed for tumor immunity induced by plasmid (HER-2 and GM-CSF) immunization. HER-2 specific tumor immunity relied completely on both CD4+ and CD8+ T cells. IFN gamma, and to a lesser extent IL-4, seemed to be crucial cytokines during tumor rejection. Protection was associated with production of anti-HER-2 IgG antibodies in B cell competent mice. After immunization, however, B cell-deficient mice rejected HER-2-expressing tumors as efficiently as control littermates. We conclude that T cells are the main effector cells in DNA vaccine induced immunity against HER-2 and that anti HER-2 antibodies are not necessary to elicit a protective anti tumor immune response in this model. PMID- 14750179 TI - MHC class I antigen processing pathway defects, ras mutations and disease stage in colorectal carcinoma. AB - Colorectal tumorigenesis has been associated with the progressive acquisition of a variety of genetic alterations. These include mutations of the Ki-ras proto oncogene in codons 12 and 13, which account for 85% of genetic changes in colorectal cancer. In murine in vitro models of oncogenic transformation, an association between ras-mediated transformation and downregulation of different components of the MHC class I antigen processing machinery (APM) has been described. In order to investigate whether this association also exists in human tumors, 10 cases of high-grade intraepithelial neoplasia (HIN), as well as primary tumors and autologous lymph node metastases from 42 patients with colorectal carcinoma, were monitored by allele-specific restriction analysis for Ki-ras mutations. In parallel, APM component expression and tumor cell proliferation were analyzed by immunohistochemistry. In comparison to autologous colorectal mucosa, TAP1, LMP2 and tapasin loss was found in 68%, 67% and 80% of HIN, respectively. In contrast, impaired TAP1, LMP2 and tapasin expression was found in 42%, 42% and 63% of primary adenocarcinomas of stage III disease and in 63%, 47% and 79% of the matched lymph node metastases, respectively. More than 60% of colorectal tumor lesions with TAP1, LMP2 and/or tapasin defects displayed Ki-ras mutations. The frequency of TAP1, LMP2 and tapasin loss varied between 33% of primary adenocarcinomas, 40% of HIN to approximately 67% of metastases. These data suggest that i) APM component deficiencies occur more frequently in Ki-ras mutated colorectal carcinoma lesions and ii) APM abnormalities in conjunction with Ki-ras mutations appear to be associated with disease stage. These findings support the hypothesis that Ki-ras mutations may contribute to immune escape mechanisms of tumors by downregulating the MHC class I APM component expression. PMID- 14750180 TI - Survival of cervix cancer patients in Harare, Zimbabwe, 1995-1997. AB - The survival experience of 284 patients with cancer of the cervix uteri registered by the population-based Zimbabwe National Cancer Registry in 1995-1997 is described. The vital status of these subjects was established by linkage with death registration and by retrieval of patient files from medical records departments. Untraced patients were contacted at home. Of the 284, 177 (62.3%) were dead and 76 (26.8%) were alive at the closing date of the study (31 December 1999), with only 31 cases (10.9%) lost to follow-up. Overall observed and relative survival at 3 years were 44.2% and 45.2%, respectively. Half of the cases (139) had been referred and treated in the radiotherapy department. Survival was significantly greater in the first 3 years for patients who received radiotherapy treatment compared to those that had not, but this difference had disappeared by the fourth year of follow-up. Many cases presented late (distant metastasis), and extent of disease was an important determinant of survival; cases with metastases had a risk of death some 3 times that of patients with localized disease. The results demonstrate the importance of earlier diagnosis and availability of effective treatment in the African context. PMID- 14750181 TI - Fibre intake and prostate cancer risk. AB - Dietary fibre has been reported to protect from several neoplasms, but the issue remains controversial. No previous study considered in depth the topic of fibres and prostate cancer. A multicentre case-control study was conducted in Italy from 1991 to 2002, including 1,294 men with incident, histologically confirmed prostate cancer and 1,451 controls admitted to the same network of hospitals as cases with acute nonmalignant conditions. Multivariate odds ratios (OR) and 95% confidence intervals (CI) were obtained after allowance for major identified confounding factors, including total energy intake. Compared to the lowest quintile, the OR of prostate cancer for the highest quintile of total fibre intake was 0.93 (95% CI 0.71-1.22). The risk was inversely related with soluble fibre (OR = 0.89, 95% CI 0.78-1.02, for a difference between 80th and 20th percentile), cellulose (OR = 0.88, 95% CI 0.78-1.01) and vegetable fibre (OR = 0.82, 95% CI 0.73-0.93). These relationships were consistent across strata of age, family history of prostate cancer, body mass index and education. Vegetable fibres appear, therefore, to have a favourable association with prostate cancer risk. PMID- 14750182 TI - Target cell-restricted and -enhanced apoptosis induction by a scFv:sTRAIL fusion protein with specificity for the pancarcinoma-associated antigen EGP2. AB - The apparent tumor selective apoptosis-inducing activity of recombinant soluble TNF-related apoptosis-inducing ligand (TRAIL) has aroused much interest for use in clinical application. However, to exploit fully its therapeutic potential, the characteristics of both the TRAIL receptor system and soluble TRAIL (sTRAIL) should be taken into account: first, the widespread expression of the various TRAIL receptors throughout the human body; second, the differential binding affinities and crosslinking requirements of the agonistic receptors TRAIL-R1 and TRAIL-R2; and third, the solution behavior of particular sTRAIL preparations. Therefore, we constructed a novel TRAIL fusion protein, designated scFvC54:sTRAIL, comprising the human scFv antibody fragment C54 genetically linked to the N-terminus of human sTRAIL. The scFvC54:sTRAIL fusion protein was designed to induce apoptosis by crosslinking of agonistic TRAIL receptors only after specific binding of scFvC54:sTRAIL to the abundantly expressed carcinoma associated cell surface antigen EGP2 (alias EpCAM). Target antigen-restricted apoptosis induction was demonstrated for various EGP2-positive tumor cells and could be inhibited by an EGP2 competing antibody. Target antigen binding converted soluble scFvC54:sTRAIL into a membrane-bound form of TRAIL that was capable of signaling apoptosis not only through TRAIL-R1 but also through TRAIL R2. Size-exclusion fast protein liquid chromatography (FPLC) indicated that scFvC54:sTRAIL was produced as stable and homogeneous trimers in the absence of detectable TRAIL aggregates. The favorable characteristics of the scFvC54:sTRAIL fusion protein potentially reduce the amount of sTRAIL required for antitumor activity and may be of value for the treatment of various human carcinomas. PMID- 14750183 TI - Effects of trastuzumab on epidermal growth factor receptor-dependent and independent human colon cancer cells. AB - Little information is available as to the potential role of HER-2 as a therapeutic target in colon cancers, which express much fewer HER-2 receptors than breast cancer cells. Treatment of certain human colon cancer cell lines with the HER-2 inhibitory antibody mAb 4D5 demonstrated a role for HER-2 in mediating proliferation, apoptosis and tumorigenicity. However, only the cell lines that were dependent on autocrine EGFR-mediated cell proliferation were susceptible to the antiproliferative and antitumorigenic effects of HER-2 inhibition. The relative levels of HER-2, EGFR, HER-3 and HER-4 were not predictive of responsiveness to mAb 4D5. Treatment with HER-2 antibodies caused a decrease in HER-2 protein levels in all of the colon cancer cell lines and also significantly decreased EGFR levels but only in the EGFR-dependent cell lines. Treatment with mAb 4D5 caused the rapid ubiquitination and ligand-dependent downregulation of the EGFR in an EGFR-dependent colon cancer cell line. Treatment of athymic mice engrafted with EGFR-dependent colon cancer cells with mAb 4D5 caused tumor regression and a decrease in EGFR tyrosine phosphorylation in the tumor cells. EGFR-independent colon cancer cell xenografts were resistant to mAb 4D5 therapy. Combined inhibition of HER-2 and EGFR caused large areas of necrosis in EGFR dependent colon cancer xenografts, suggesting a benefit of combined HER-2 and EGFR inhibitor therapy. Predicting clinical responsiveness of human colon cancer cells to anti-HER-2 and anti-EGFR therapy may require demonstration of EGFR tyrosine kinase dependency of the cells. PMID- 14750184 TI - Proliferation assay on a silicon chip applicable for tumors extirpated from mammalians. AB - We describe a novel anticancer drug sensitivity assay on a silicon chip applicable for tumors extirpated from in vivo mammalians. Human promyelocytic leukemia (HL-60) cells were subcutaneously (s.c.) inoculated in SCID mice, then removed 31 days after the inoculation. The cells were embedded in a small volume (18 nL) of a collagen-gel matrix on a pyramid-shaped silicon microstructure for further cultivation. The respiration activity of the cells on the chip was measured by scanning electrochemical microscopy (SECM). The proliferation behavior was continuously monitored for 6 days. It seemed that the proliferation rate of the cells removed from the mice was lower than that cultured in a flask and conformed to that in mice. The effects of cisplatin (CDDP) and etoposide (VP 16) on the HL-60 cultured in vivo were in good agreement with those obtained by a conventional colorimetric assay. Our results suggest that the SECM-based assay is appropriate for biopsy specimens in a relatively short-time evaluation. PMID- 14750185 TI - Molecular analysis of PinX1 in medulloblastomas. AB - Our group has previously demonstrated a high frequency of allelic loss on the short arm of chromosome 8 and identified a region of homozygous deletion of 1.41 Mb, flanked by D8S520 and D8S1130, on 8p23.1 in medulloblastomas, suggesting the presence of a tumor suppressor gene in this critical deletion region. The aim of our study was to investigate whether PinX1, a newly identified gene whose product is a potent inhibitor of telomerase, is the target gene in the homozygous deletion region identified in medulloblastomas. We assessed for alterations in gene sequence and transcript expression of PinX1, as well as the correlation between PinX1 expression and telomerase activity in a series of 52 medulloblastomas, 3 medulloblastoma cell lines (D283, D341 and Daoy) and 4 primitive neuroectodermal tumors (PNETs). Direct sequence analysis of all 7 exons and splice junctions of the PinX1 gene revealed no somatic mutations but 11 genetic polymorphisms. Transcript expression of PinX1, as evaluated by reverse transcription-polymerase chain reaction, in microdissected tumors and normal cerebellum showed 2 transcript variants, corresponding to the full-length form and an alternative spliced variant lacking exon 6, in all samples. This result indicated that PinX1 expression was not suppressed in medulloblastomas. Using the telomeric repeat amplification protocol (TRAP) assay, 13 of 19 (68%) medulloblastomas, 1 of 2 PNETs and all 3 cell lines showed telomerase activity. There is no significant correlation between PinX1 transcript expression and telomerase activity, but our results showed that telomerase activation is involved in medulloblastomas. Taken together, our results suggest that PinX1 does not play a major role in the oncogenesis of medulloblastomas. PMID- 14750187 TI - In memory of Professor Sandy C. Marks, Jr., B.S., D.D.S., Ph.D. (1937-2002). PMID- 14750188 TI - A social biography of Carnegie embryo no. 836. AB - A tiny, sectioned embryo specimen known as Carnegie no. 836 has served as the prototype for Stage 13 (28-32 days) since the 1910s. Recently digitalized and reanimated for the 21st century, this singular specimen is now being used to develop 3D and 4D visualizations. Yet the social origins of the specimen have been largely forgotten. This essay traces the biography of 836 from its origins in a young woman's life, through sectioning and transformation into a scientific specimen, to its contemporary manifestations as a symbol of life. By reuniting the specimen with its story, we can appreciate how cultural attitudes toward embryo specimens have changed over the past century. PMID- 14750189 TI - Gunther von Hagens and Body Worlds Part 1: the anatomist as prosektor and proplastiker. AB - Recent calls to reintegrate the sciences and humanities are challenged by the contemporary work of anatomist Gunther von Hagens and his Body Worlds exhibits of plastinated cadavers. The anatomical quest to understand our physical interior has long been in tension both with aesthetic ideals and religious sensitivities regarding the metaphysical significance of the human body. Part I of this two part Historical Note examines tensions epitomized by Goethe's figures of the prosektor and proplastiker. The former, driven by scientific curiosity, is willing to destroy, even desecrate, the human form to obtain knowledge. The latter demurs at such mutilation of our physical body, wondrous even in death- seeking instead to rejoin what the prosektor has pulled apart, to restore human dignity. In the confrontation between prosektor and proplastiker, roles disturbingly fused in the person of von Hagens himself, questions arise regarding the authenticity of models as well as the appropriate recipients of such mediated yet intimate anatomical knowledge. Part II will focus on religious perspectives on the human body, variously interpreted as God's handiwork, habitation for the soul, and vehicle of resurrection. Consideration also is given to the role of anatomist as priest, prophet, and Promethean creator, roles self-consciously embraced by von Hagens. PMID- 14750190 TI - Preserving and sharing examples of anatomical variation and developmental anomalies via photorealistic virtual reality. AB - Computer graphics technology has made it possible to create photographic-quality virtual specimens from real anatomical material. One technique for doing this, QuickTime Virtual Reality (QTVR), results in virtual specimens that are easily shared on the Internet and displayed as standalone entities or incorporated into complex programs or Web sites. A compelling use of this technology is the sharing of rare specimens such as unusual variations, developmental anomalies or gross pathology. These types of specimens have traditionally been confined to anatomical museums, but could serve a much more useful existence as freely shared virtual specimens. An example presented here is a relatively rare developmental defect in the embryonic aortic arches that results in a right-sided aortic arch coursing posterior to the trachea and esophagus. In a time of ever increasing restraints on the practical side of anatomy education, an Internet-based library of human variation and other rare specimens would be a useful supplement to students' limited exposure to the human body. Since the discovery and preparation of specimens would be the rate-limiting step in producing such a collection, we propose the establishment of a center for virtual specimen creation and preservation through a cooperative effort by gross anatomists and pathologists in contributing the source material. This collection, a work in progress, is available at www.anatomy.wright.edu/qtvr. PMID- 14750191 TI - What happened in the origin of human consciousness? AB - At some point in its evolutionary history, our species Homo sapiens ceased to be a nonlinguistic, nonsymbolic organism, living in the world as presented to it by Nature, and instead began to exist in a world that it reconstructs in its own mind. Most scientists since Darwin have been content to explain this extraordinary transformation in human consciousness by the operation of natural selection. However, the human fossil and archaeological records indicate that modern human symbolic consciousness is not the culmination of the long trend that natural selection would predict. Instead, it shows that major change in the human past has been episodic and rare and that, as far as can be determined from the archaeological record, the passage from nonsymbolic to symbolic cognition is a recent event as well as an unprecedented one. So recent, indeed, that it significantly postdates the acquisition of modern human anatomy as expressed in skeletal structure. It, thus, appears most likely that the biological (neural) capacity underwriting the radically new behavioral mode arose as an incidental exaptation in the same process that produced the new skeletal structure of Homo sapiens, but that it lay unexpressed until it was "discovered" by means of a cultural innovation, plausibly the invention of language. As in the case of the modern anatomical structure, it appears that the new capacity was initially expressed in Africa and that its various behavioral potentials were sequentially discovered in a drawn-out process that is continuing today. An "accidental" origin of the human capacity helps understand why so many human behaviors have proven self-destructive and contradictory, a feature of our species that reductionist, selection-based scenarios are hard-put to explain. PMID- 14750192 TI - Active components from Artemisia iwayomogi displaying ONOO(-) scavenging activity. AB - Peroxynitrite (ONOO(-)) is one of cytotoxic species produced by the reaction between superoxide (*O(2) (-)) and nitric oxide (NO). The main aim of this study was to identify ONOO(-) scavenging constituents from herbs. Methanolic extracts derived from one hundred thirty eight herbs were screened out for their ONOO(-) scavenging activities. Twenty herbs showed excellent ONOO(-) scavenging activity in a dose dependent manner. Of them, Artemisia iwayomogi, which showed the highest activity, was fractioned with several solvents. The ONOO(-) scavenging activity of fractions was in order of ethyl acetate (EtOAc) > n-butanol (BuOH) > dichloromethane (CH(2)Cl(2)) > water (H(2)O) fraction. The EtOAc and the n-BuOH fractions were further purified to isolate active principles by column chromatography. Apigenin 7-methylether (genkwanin), apigenin 7,4'-di-O methylether, jaceosidin, and scopoletin from the EtOAc fraction and chlorogenic acid, 2,4-dihydroxy 6-methoxy acetophenone 4-O-beta-D-glucoside, quebrachitol, and scopolin from the n-BuOH fraction were identified. The scavenging activities of chlorogenic acid (IC(50) = 0.52 +/- 0.04 microM), genkwanin (IC(50) = 1.01 +/- 0.10 microM), and scopoletin (IC(50) = 1.03 +/- 0.15 micro M) were higher than or comparable to a well-known ONOO(-) scavenger, penicillamine (IC(50) = 1.76 +/- 0.18 microM), suggesting that those compounds might be developed as effective ONOO(-) scavengers for, prevention of the ONOO(-)-involved diseases. PMID- 14750193 TI - Reproduction in male rats is vulnerable to treatment with the flavonoid-rich seed extracts of Vitex negundo. AB - A partially purified flavonoid-rich extract was prepared from the seed of Vitex negundo. The effect of this extract on the reproductive system of male rats was investigated at four different concentrations. All the major accessory sex organs shed weight when the preparation was administered at doses of >or=15 mg/rat/day after 15 days of treatment. The drop in weight was also reflected in disturbed tissue biochemistry. Secretory products such as citric acid in the prostate, fructose in seminal vesicles and epididymal alpha-glucosidase activity, indices of accessory sex organ function in males, diminished. Microscopic examination of the sperm derived from the cauda epididymides of treated animals showed only a marginal change in vitality. However, sperm numbers dwindled and slackness in their motility was observed, factors that may impede fertility. Toxicity testing in blood did not point to distress in any of the vital organs. Taken together, it is inferred that the seed extracts of V. negundo interfere with male reproductive function without producing adverse toxicity in other vital organs. PMID- 14750194 TI - The evaluation of the radioprotective effect of chyavanaprasha (an ayurvedic rasayana drug) in mice exposed to lethal dose of gamma-radiation: a preliminary study. AB - The effect of various doses of 50% ethanolic extract of chyavanaprasha (an Ayurvedic rejuvenating herbal preparation) was studied on the survival of mice exposed to 10 Gy of gamma-radiation. Treatment with chyavanaprasha, consecutively for fi ve days before irradiation, delayed symptoms of radiation sickness and onset of mortality when compared with the non-drug treated irradiated controls. All doses of chyavanaprasha provided a significant protection against gastrointestinal (GI) death (death of animals within 10 days after exposure to radiation), however, highest protection against GI death was observed for 15 mg/kg chyavanaprasha. Chyavanaprasha also provided a significant protection against the bone marrow death after 10 to 40 mg/kg. However, the best protection was seen for 15 mg/kg, where the highest number of survivors was observed at the end of 30 days post-irradiation. The drug was non-toxic up to a dose of 6 g/kg b. wt., the highest drug dose that could be tested. Our study demonstrates that chyavanaprasha can provide good radioprotection at a very low non-toxic dose. PMID- 14750195 TI - Effects of methanol extract of Cirsium japonicum var. ussuriense and its principle, hispidulin-7-O-neohesperidoside on hepatic alcohol-metabolizing enzymes and lipid peroxidation in ethanol-treated rats. AB - Effects of the methanol extract of Cirsium japonicum var. ussuriense and hispidulin 7-O-neohesperidoside isolated from the plant on hepatic alcohol metabolizing enzymes and lipid peroxidation were studied in rats treated with ethanol. Rats treated with 10% alcohol solution for 6 weeks were orally administered with 250 or 500 mg of methanol extract or 10 or 20 mg of hispidulin 7-O-neohesperidoside per kg body weight daily during the last week of ethanol treatment. The administration of the methanol extract of herbal plant and hispidulin 7-O-neohesperidoside in ethanol-treated rats significantly enhanced the activities of hepatic alcohol dehydrogenase, microsomal ethanol-oxidizing system and aldehyde dehydrogenase in a dose-dependent manner. The extract and the compound decreased hepatic lipid peroxidation along with an increase in hepatic content of reduced glutathione. The methanol extract and hispidulin 7-O neohesperidoside of C. japonicum var. ussuriense also increased the activity of glutathione reductase, but had no effect on gamma-glutamylcysteine synthase. The results suggest that C. japonicum var. ussuriense may alleviate alcoholic toxicity by enhancing ethanol oxidation as well as inhibiting lipid peroxidation, and hispidulin 7-O-neohesperidoside is one of the active substances responsible for the protective effects of this plant. PMID- 14750196 TI - Antiosteoporotic activity of OST-6(Osteocare), a herbomineral preparation in calcium deficient ovariectomized rats. AB - The preventive effect of the herbal formulation OST-6 (Osteocare) on the progress of bone loss induced by ovariectomy and concurrent calcium deficiency was studied in rats. Ovariectomy (OVX) and calcium deficiency (CD) resulted in bone loss as evident from decreased femoral weight and density. Treatment with OST-6 at a dose of 250 mg/kg body wt twice a day orally for a period of 16 weeks significantly restored the femoral weight and density. The biomechanical strength of the tibia was decreased by OVX and CD and this was significantly suppressed by the administration of OST-6. Histologically decalcified bone sections revealed narrowed, and the disappearance of, trabeculae and widened medullary spaces. The total numbers of tartrate-resistant acid phosphates (TRAP) positive cells were increased significantly in OVX animals. Treatment with OST-6 significantly inhibited these histopathological changes and strongly suggested that OST-6 was effective in preventing the progress of bone loss induced by ovariectomy and concurrent calcium deficiency. PMID- 14750197 TI - Chemical and biological evaluation of the essential oils of different Melaleuca species. AB - The essential oils of the fresh leaves of M. ericifolia, M. leucadendron, M. armillaris and M. styphelioides were isolated by a hydrodistillation method and analysed by a gas chromatography/mass spectrometry (GC/MS) technique. The essential oil of M. ericifolia contained methyl eugenol (96.84%) as a major constituent, whereas M. leucadendron was rich in 1,8-cineole (64.30%). The essential oil of M. armillaris was rich in 1,8-cineole (33.93%) followed by terpinen-4-ol (18.79%), whereas M. styphelioides was rich in caryophyllene oxide (43.78%) and (-) spathulenol (9.65%). The essential oils of these species possessed antimicrobial and antifungal activities. M. ericifolia exhibited the highest inhibitory effects against Bacillus subtiles and Aspergillus niger. The antiviral activities of the essential oils against Herpes simplex virus type 1 (HSV-1) were studied in African green monkey kidney cells (Vero) by a plaque reduction assay. The volatile oil of M. armillaris was more effective as a virucidal (up to 99%) than that of M. leucadendron (92%) and M. ericifolia (91.5%). The effects of the essential oils on the antioxidant system status in carbon tetrachloride treated animals were studied. The essential oil of M. armillaris exhibited a marked antioxidant effect, it improved vitamin E, vitamin C and superoxide dismutase parameters so it can be used as a free radical suppressor. PMID- 14750198 TI - Vasodilating properties of the stem bark extract of Mitragyna ciliata in rats and guinea pigs. AB - Mitragyna ciliata is commonly used in traditional medicine for the treatment of inflammation, hypertension, headache, rheumatism, gonorrhoea and broncho pulmonary diseases. In the present study, the vascular relaxant effect in the rat and guinea-pig was investigated. The extract induced aortic relaxation in a concentration-dependent manner, with an EC(50) of 1.3 and 7 microg/mL for the noradrenaline- and KCl-induced contractions, respectively. The relaxant effect of the extract on KCl-induced contractions was fi ve times greater than on noradrenaline-induced contractions. Moreover, the relaxant effect of the extract was higher in rat aortic rings with endothelium (104.67%) than without endothelium (49.44%). Chemical analysis of the extract showed the presence of alkaloids and flavonoids which may be responsible for the antihypertensive properties. PMID- 14750199 TI - Chemical analysis and antifungal activity of Thymus striatus. AB - The essential oil composition from Thymus striatus collected from Mountain Orjen (Montenegro) has been investigated by gas chromatography-mass spectrometry. Thymol, gamma-terpinene and p-cymene were found to be the major components. Furthermore, the oil and its major component, thymol, were analysed for potential antifungal activity against plant, animal and human pathogenic fungi from different genera by a macrodilution test. The oil exhibited a strong inhibitory effect against all fungi investigated. PMID- 14750200 TI - Methanol extract of the oyster mushroom, Pleurotus florida, inhibits inflammation and platelet aggregation. AB - The antiinflammatory and platelet aggregation inhibiting activities of the methanol extract of Pleurotus florida Eger, an edible and commercially grown mushroom, were investigated. The extract showed significant activity in ameliorating acute inflammation induced by carrageenan and chronic inflammation by formalin at 500 and 1000 mg/kg body weight. The effect was comparable to the standard reference drug, diclofenac. The extract also showed significant platelet aggregation inhibiting activity of washed human platelets. Adenosine 5' diphosphate (ADP) induced platelet aggregation was inhibited by 88% to 95% at a concentration of 500 microg/mL of the extract after a preincubation time of 5, 10 and 20 min. The marked antiinflammatory and platelet aggregation inhibiting properties of this mushroom suggest its potential therapeutic use against vascular disorders. PMID- 14750201 TI - Comparative controlled study of Andrographis paniculata fixed combination, Kan Jang and an Echinacea preparation as adjuvant, in the treatment of uncomplicated respiratory disease in children. AB - A three-arm study comparing the efficacy of Kan Jang, a fixed herbal combination containing standardized Andrographis paniculata (N.) SHA-10 extract, with Immunal, a preparation containing Echinacea purpurea (L.) extract, in uncomplicated common colds was carried out in 130 children aged between 4 and 11 years over a period of 10 days. The study was designed as an adjuvant treatment of Kan Jang and Immunal with a standard treatment. The patients were assigned to one of the three groups. In control group C; 39 patients received only standard treatment. Kan Jang and Immunal were used as an adjuvant to this therapy in the other two groups. Adjuvant group A; 53 patients treated with Kan Jang tablets concomitant to standard treatment, and adjuvant control group B; 41 patients treated with concomitant Immunal. It was found that the adjuvant treatment with Kan Jang, was significantly more effective than Immunal, when started at an early stage of uncomplicated common colds. The symptoms of the disease were less severe in the Kan Jang group. The effect of Kan Jang was particularly pronounced in two objective parameters, amount of nasal secretion g/day and nasal congestion. Kan Jang also accelerated the recovery time, whereas Immunal did not show the same efficacy. The use of standard medication was significantly less in the Kan Jang adjuvant group than in either the Immunal or standard treatment group. Kan Jang treatment was well tolerated and no side effects or adverse reactions were reported. PMID- 14750202 TI - Emblica officinalis causes myocardial adaptation and protects against oxidative stress in ischemic-reperfusion injury in rats. AB - The fruits of Emblica officinalis (Amla) are widely used in the Indian System of Medicine and are believed to increase defense against disease. In the present study, the effects of chronic oral administration of fresh fruit homogenate of Amla on: (i). myocardial antioxidant system and (ii). oxidative stress induced by ischemic-reperfusion injury (IRI) in rat heart were investigated. Fresh amla fruit homogenate, in three different doses (250, 500 and 750 mg/kg) and normal saline (C) were administered orally to Wistar albino rats (120-150 gms) of either sex daily for 30 days. There was reduction in basal myocardial lipid peroxidation, as evidenced by decreased thiobarbituric acid reactive substances (TBARS) level, and augmentation of myocardial endogenous antioxidants, like superoxide dismutase (SOD), catalase and glutathione peroxidase (GPx) in the treated rats. Hearts were also subjected to in vitro IRI (9 min of global ischemia, followed by 12 min of reperfusion, Langendorff's mode). Significant myocyte injury and rise in myocardial TBARS along with depletion of SOD, catalase, GSH (reduced glutathione) and GPx occurred in the control group. No significant increase in myocardial TBARS and depletion of antioxidant enzymes were observed in the treated groups. Myocyte injury was evident only in 250 mg/kg group. The results indicate that chronic Emblica officinalis administration causes myocardial adaptation by augmenting endogenous antioxidants and protects rat hearts from oxidative stress associated with ischemic-reperfusion injury. PMID- 14750204 TI - Cytotoxic activity of Hypericum perforatum L. on K562 erythroleukemic cells: differential effects between methanolic extract and hypericin. AB - The influence of a methanolic extract of Hypericum perforatum L. and of purified hypericin has been comparatively tested on the growth of a human erythroleukemic cell line (K562). After 1 h exposure to increasing concentrations (as hypericin content) of both agents in the dark, leukemic cells were grown for 24 h and 48 h. The effects on cell growth were determined by viable cell count, flow cytometry analysis and fluorescence microscopy. Our data show that purified hypericin has only a weak inhibitory effect on cell growth and no effect in inducing apoptotic cell death. In contrast, the Hypericum flower extract shows a significant concentration-dependent and long-lasting inhibition of cell growth, and induces apoptotic cell death. This work con fi rms the interesting role of Hypericum perforatum L. in cancer therapy and strongly supports the hypothesis that agents, other than hypericin, present in the total extract can impair tumor cell growth acting separately or in a combined manner. PMID- 14750203 TI - Inhibitory activity of flavonoids from Lychnophora sp. on generation of reactive oxygen species by neutrophils upon stimulation by immune complexes. AB - Formation of circulating immune complexes (ICs) is essential for clearance of invading agents. In some circumstances ICs might deposit on host tissues, leading to an inflammatory process that involves massive activation of neutrophils (PMNs), release of reactive oxygen species (ROS) and lysosomal enzymes and damage to the host tissue. Extracts of plants from Lychnophora sp. are used in Brazilian folk medicine as antiinflammatory agents. In this study, we evaluated the effect of eight flavonoids isolated from L. granmongolense, L. salicifolia and L. ericoides on the generation of ROS by rabbit PMNs stimulated with two kinds of ICs: particles of serum-opsonized zymosan (OZ) and insoluble ICs (ICIgG). ROS production was measured by chemiluminescence (CL) assay. We observed that 5- and 7- dihydroxylated compounds at 5 micromol/L inhibited almost totally ICIgG- and OZ-triggered luminol-CL and OZ-triggered lucigenin-CL. The degree of inhibitory effect among the other flavonoids was different, depending on the kind of ICs used to trigger ROS generation by PMNs and the number and position of methoxy groups. Moreover, under the conditions assessed, the studied flavonoids were not toxic to the rabbit PMNs. These results suggest that the actions of flavonoids on ROS generation by stimulated PMNs are highly dependent on their structures. PMID- 14750205 TI - The effect of medicinal plants of Islamabad and Murree region of Pakistan on insulin secretion from INS-1 cells. AB - In vitro testing of the extracts of medicinal plants collected from Islamabad and the Murree region on insulin secretagogue activity was carried out. Dried ethanol extracts of all plants (ZH1-ZH19) were dissolved in ethanol and DMSO, and tested at various concentrations (between 1 and 40 microg/mL) for insulin release from INS-1 cells in the presence of 5.5 mM glucose. Glibenclamide was used as a control. Promising insulin secretagogue activity in various plant extracts at 1, 10, 20 and 40 microg/mL was found, while in some cases a decrease in insulin secretion was also observed. Artemisia roxburghiana, Salvia coccinia and Monstera deliciosa showed insulin secretagogue activity at 1 microg/mL (p < 0.05) while Abies pindrow, Centaurea iberica and Euphorbia helioscopia were active at 10 microg/mL (p < 0.05). Extracts of Bauhinia variegata and Bergenia himalacia showed effects at 20 microg/mL (p < 0.05), and Taraxacum officinale and Viburnum foetens at 40 microg/mL (p < 0.05). Insulin secretagogue activity could not be detected in the extracts of Adhatoda vasica, Cassia fistula, Chrysanthemum leucanthemum, Morus alba, Plectranthus rugosus, Peganum harmala and Olea ferruginea. The results suggest that medicinal plants of Islamabad and the Murree region of Pakistan may be potential natural resources for antidiabetic compounds. PMID- 14750207 TI - Effects of Rhus coriaria L (Anacardiaceae) on lipid peroxidation and free radical scavenging activity. AB - In order to evaluate the antioxidant potential of Rhus coriaria L. (Anacardiaceae), a well-known spice, the methanolic extract (water-soluble part) was prepared and investigated using free radical-generating systems in vitro. The IC(50) value of the extract for lipid peroxidation was estimated as 1200 microg/mL in the Fe(+2)-ascorbate system while those for superoxide-scavenging activity in the xanthine-xanthine oxidase method and hydroxyl radical scavenging activity in the deoxyribose decomposition method were 282.92 microg/mL and 3850 microg/mL, respectively. These data suggest that the methanolic extracts of Rhus coriaria L. fruits have considerable antioxidant activity against free radicals and lipid peroxidation in vitro, a fact that may encourage in vivo studies. PMID- 14750206 TI - Anti-inflammatory activity, cytotoxicity and active compounds of Tinospora smilacina Benth. AB - Tinospora smilacina Benth. has been used in Australian indigenous medicine for the treatment of headache, rheumatoid arthritis and other inflammatory disorders. As part of an investigation into the anti-inflammatory potential of plants using an ethnopharmacological approach, the present study sought to evaluate the efficacy and safety of Tinospora smilacina. An ethanol extract of this plant was evaluated in vitro for anti-inflammatory activities on cyclooxygenase-1 (COX-1), cyclooxygenase-2 (COX-2), 5-lipoxygenase (5-LO) and phospholipase A(2) (PA(2)). The ethanol extract of Tinospora smilacina showed inhibitory activities on COX-1, COX-2, 5-LO and PA(2) with the IC(50) values of 63.5, 81.2, 92.1 and 30.5 micro g/mL respectively. Cytotoxic effect of the extracts of Tinospora smilacina was investigated in vitro using ATP-based luminescence assay and the results showed no cytotoxic effect on cell lines of skin fibroblasts (1BR3), human Caucasian hepatocyte carcinoma (Hep G2) and human Caucasian promyelocytic leukaemia (HL 60). This paper also describes the results of fractionations and bioassay guided chemical studies, suggesting that the anti-inflammatory activity is due to triterpene-fatty acid esters and free fatty acids. PMID- 14750208 TI - Effect of garlic oil on ethanol induced gastric ulcers in rats. AB - Garlic oil was evaluated for gastroprotective activity against ethanol induced ulcers. Reactive oxygen species are involved in the pathogenesis of these ulcers. The possible involvement of garlic oil in restraining the oxidation process produced in gastric tissue was also investigated. The ulcer index, lipid peroxidation and antioxidant enzyme activity (GPx, catalase, SOD) were determined. Pretreatment with garlic oil in doses of 0.25 and 0.5 mg/kg, 30 min before administration of ethanol (1 mL of 100%) caused a decrease in ulcer index and lipid peroxidation and ameliorated the decrease in antioxidant enzyme levels caused by ethanol. The result suggests that garlic oil possesses antioxidant properties and provides protection against ethanol induced gastric injury. PMID- 14750209 TI - Antiproliferative activity of Copaifera duckei oleoresin on liver regeneration in rats. AB - Copaiba oleoresin is widely used in folk medicine as an antiinflammatory, healing and antiseptic agent. Wistar rats received Copaifera duckei oleoresin (CDO) or saline solution (SS) from 7 days before surgery until killing. Partial hepatectomy (PH) or sham-operation was performed. Hepatocellular proliferation and liver mitochondrial respiration were evaluated 24 h after the procedure. The proliferating cell nuclear antigen labelling index was significantly lower in CDO treated plus PH rats than in SS-treated plus PH rats, suggesting that CDO inhibited cell proliferation. The state 3 and state 4 respiration rates were greater for CDO-treated groups, and the stimulation of state 4 respiration was more expressive. These data resulted in a significant decrease in the respiratory control ratio observed for CDO-treated groups, demonstrating the ability of CDO to uncouple oxidative phosphorylation in mitochondria, a fact that may be one explanation for the inhibition of hepatocellular proliferation observed in the CDO-treated plus PH group. PMID- 14750210 TI - Isolation and purification of the hypoglycaemic principle present in Capsicum frutescens. AB - Capsicum frutescens has been used to treat diabetes mellitus by traditional healers in Jamaica. Purification experiments employing thin layer chromatography (TLC) and high performance liquid chromatography led to the extraction of the active principle, capsaicin. Purified capsaicin caused a decrease in blood glucose levels to 4.91 +/- 0.52 (n = 6) mmol/dL versus 6.40 +/- 0.13 mmol/dL (n = 6) for the control (p < 0.05) at 2.5 h in an OGTT in dogs. There was a concomitant elevation in plasma insulin levels (p < 0.05). In conclusion, it can be stated that capsaicin is the major constituent of Capsicum frutescens that is responsible for the hypoglycaemic episodes seen in the dogs. It is also apparent that the latter is mediated by insulin release. PMID- 14750211 TI - Eighteenth Aspen Cancer Conference: mechanisms of toxicity, carcinogenesis, cancer prevention, and cancer therapy. PMID- 14750212 TI - Modulation of DNA hypomethylation as a surrogate endpoint biomarker for chemoprevention of colon cancer. AB - Surrogate end-point biomarkers are being developed as indicators of the efficacy of chemopreventive agents. These biomarkers are molecular and biological end points that can be modulated by chemopreventive agents in accordance with their efficacy to prevent cancer. DNA hypomethylation is a common alteration found in colon tumors that has the potential of being modulated by chemopreventive agents and thus being useful as a surrogate end-point biomarker. Agents that were either effective or ineffective in preventing colon cancer were evaluated for the ability to modulate DNA hypomethylation in azoxymethane-induced colon tumors in male F344 rats. DNA methylation was determined by Dot Blot Analysis using a mouse monoclonal anti-5-methylcytosine antibody. Colon tumors had a 70% reduction in DNA methylation relative to normal colonic mucosa. DNA methylation in the tumors was increased by 7 days of treatment with agents that have been shown to prevent colon cancer (calcium chloride, alpha-diflouromethylornithine [DFMO], piroxicam, and sulindac), whereas agents shown not to prevent colon cancer in rats (low dose aspirin, 2-carboxyphenyl retinamide [2-CPR], quercetin, 9-cis retinoic acid, and rutin) did not increase DNA methylation. The results suggest that the ability to reverse the DNA hypomethylation in colon tumors could be useful as a surrogate end-point biomarker for chemoprevention of colon cancer. PMID- 14750214 TI - Suppression of polyamine catabolism by activated Ki-ras in human colon cancer cells. AB - An activated Ki-ras was expressed in the human colon adenocarcinoma cell line Caco-2 to study the effects of Ki-ras oncogene on polyamine metabolism during gastrointestinal tumorigenesis. Multiple clones selected for expression of the mutant Ki-ras transgene displayed a suppression of transcription of a key catabolic enzyme in polyamine catabolism spermidine/spermine N1-acetyltransferase (SSAT). Gene expression analysis, with cDNA microarrays, showed that Ki-ras transfected clones had decreased levels of expression, compared to mock transfected cells, of peroxisome proliferator-activated receptor gamma (PPARgamma), a member of the nuclear hormone receptor family and an important regulator of cell proliferation and differentiation. The activated Ki-ras suppressed SSAT expression by a mechanism involving the PPARgamma response element (PPRE) located at +48 bp relative to the transcription start site of the SSAT gene. Transient expression of the PPARgamma protein in Ki-ras expressing Caco-2 clones, or treatment with the PPARgamma ligand ciglitazone, led to an increase in the SSAT promoter activity. A MEK1/2 inhibitor PD98059 induced transcription of both PPARgamma and SSAT genes in the activated Ki-ras clones, suggesting that the mitogen-activated protein kinases (MAPKs) were involved in the regulation of SSAT expression by PPARgamma. We concluded that mutated Ki-ras suppressed SSAT via a transcriptional mechanism involving the PPARgamma signaling pathway. PMID- 14750213 TI - Kinetics of myc-max-mad gene expression during hepatocyte proliferation in vivo: Differential regulation of mad family and stress-mediated induction of c-myc. AB - Mad proteins (Mad1, Mxi1, Mad3, Mad4, Mnt/Rox) are biochemical and biological antagonists of c-Myc oncoprotein. Mad-Max dimers repress the transcription of the same target genes activated by Myc-Max dimers. Despite the critical role of Max and Mad proteins as modulators of c-Myc functions, there are no comparative data on their regulation in vivo. We carried out a systematic analysis of c-myc, max, and mad family expression in a model of synchronized cell proliferation in vivo in adult tissues, that is, rat hepatocytes after partial hepatectomy. We confirmed the previously reported early peak of c-myc expression after hepatectomy but we show that it did not correlate with hepatocyte proliferation as it also occurred in sham-operated animals as a result of surgical stresses. A second peak of c-myc expression was observed later, at the time of the wave of DNA synthesis. No such expression was detected in sham-operated rat quiescent hepatocytes. max expression increased around 4-16 h after hepatectomy, before the peaks of c-myc and DNA synthesis. mxi1 and mad4 were slightly downregulated during liver regeneration. mnt/rox expression did not change. These expression patterns suggest a role of Myc-Max for efficient mitogenic response of hepatocytes. We also analyzed the effects of Myc and Max ectopic expression on the clonogenic growth of the rat hepatoma cells. Expression of c-Myc and Max increased clonogenic growth, whereas the reduction of c-Myc levels by an antisense vector decreased growth. The results suggest nonredundant roles for mad genes in hepatocyte proliferation and point to c-Myc as a putative target for anticancer therapy of liver cancer. PMID- 14750215 TI - Imbalance of antioxidant enzymes in tumor cells and inhibition of proliferation and malignant features by scavenging hydrogen peroxide. AB - The aim of this study was to evaluate the endogenous alterations of the antioxidant enzymes in tumor cells and to specifically compensate the resulting changes in the levels of reactive oxygen species (ROS) to control the malignant growth. We determined and compared the activities of antioxidant enzymes and the levels of superoxide anion (O2*-) and hydrogen peroxide (H2O2) in tumor cell lines with different degrees of malignancy, paired with regard to their origin (PB/CH72T4, PDV/PDVC57, and HBL-100/MCF-7). An increase in superoxide dismutase activity and a decrease in the activities of H2O2-detoxifying enzymes, as a function of malignancy, coupled with a rise in H2O2 and a decrease in O2*- were demonstrated. Treatment of cells with exogenous catalase showed a dose-dependent inhibition of proliferation. This inhibition was also demonstrated in several cell lines of different tissue origin and species, suggesting a general role of H2O2 in cell proliferation. Moreover, stable expression of human catalase in MCF 7 cells inhibited proliferation and also reverted malignant features. We conclude that H2O2 played a crucial and general role in the regulation of proliferation and that an endogenous imbalance in antioxidant enzymes could be a relevant event in the carcinogenesis process. PMID- 14750216 TI - Molecular mechanisms for apigenin-induced cell-cycle arrest and apoptosis of hormone refractory human prostate carcinoma DU145 cells. AB - Development of effective agents for treatment of hormone-refractory prostate cancer has become a national medical priority. We have reported recently that apigenin (4',5,7-trihydroxyflavone), found in many common fruits and vegetables, has shown remarkable effects in inhibiting cell growth and inducing apoptosis in many human prostate carcinoma cells. Here we demonstrate the molecular mechanism of inhibitory action of apigenin on androgen-refractory human prostate carcinoma DU145 cells that have mutations in the tumor suppressor gene p53 and pRb. Treatment of cells with apigenin resulted in a dose- and time-dependent inhibition of growth, colony formation, and G1 phase arrest of the cell cycle. This effect was associated with a marked decrease in the protein expression of cyclin D1, D2, and E and their activating partner, cyclin-dependent kinase (cdk)2, 4, and 6, with concomitant upregulation of WAF1/p21, KIP1/p27, INK4a/p16, and INK4c/p18. The induction of WAF1/p21 and its growth inhibitory effects by apigenin appears to be independent of p53 and pRb status of these cells. Apigenin treatment also resulted in alteration in Bax/Bcl2 ratio in favor of apoptosis, which was associated with the release of cytochrome c and induction of apoptotic protease-activating factor-1 (Apaf-1). This effect was found to result in a significant increase in cleaved fragments of caspase-9, -3, and poly(ADP-ribose) polymerase (PARP). Further, apigenin treatment resulted in downmodulation of the constitutive expression of nuclear factor-kappaB (NF-kappaB)/p65 and NF kappaB/p50 in the nuclear fraction that correlated with an increase in the expression of IkappaB-alpha (IkappaBalpha) in the cytosol. Taken together, we concluded that molecular mechanisms during apigenin-mediated growth inhibition and induction of apoptosis in DU145 cells was due to (1) modulation in cell-cycle machinery, (2) disruption of mitochondrial function, and (3) NF-kappaB inhibition. PMID- 14750217 TI - Nose and blood pressure. PMID- 14750218 TI - Slowing Parkinson's disease progression: recent dopamine agonist trials. PMID- 14750219 TI - Nelfinavir safe and effective in HIV/HCV patients. PMID- 14750220 TI - Bacteria more resistant to antibiotics. PMID- 14750221 TI - Results in for first phase III vaccine trial. PMID- 14750222 TI - Fast track for antiretroviral. PMID- 14750223 TI - Laughter is the best medicine: the value of humour in current nursing practice. PMID- 14750224 TI - A study of hyperbilirubinemia and the effect of phototherapy among full term newborns with a view to develop a nursing care protocol based on identified needs. PMID- 14750225 TI - Violence against women. PMID- 14750226 TI - [Tropical endemic limbo-conjunctivitis in Lubumbashi, Democratic Republic of the Congo]. AB - PURPOSE: To determine the frequency and clinical characteristics of tropical endemic limbo-conjunctivitis (TELC). METHODS: A review of records of all patients aged 0-15 years with a diagnosis of tropical endemic limbo-conjunctivitis examined between January 2000 and February 2002. The diagnosis was based on interview and routine ocular examination. RESULTS: A total of 139 patients out of 422 patients had the diagnosis of TELC, giving a relative frequency of 32.9%. The average age with TELC was 6.5 years which was lower than that of patients without TELC (7.6 years). The sex distribution didn't show any difference between the two sexes (P = 0.345). Children aged 0-4 years were the most affected by the disease. Itching (86%), red eye (51%), brown eye (48%), and tearing (48%) were the most frequent complaints encountered. Photophobia was only seen in 12% of patients. The distribution of the types of the disease was: palpebral form (58%), mixed form (39%) and limbal form (3%). CONCLUSION: One out of three children 0-15 years of age who consulted suffers from LCET in Lubumbashi; although other African studies reported difference in distribution in sex and the predominance of the limbal (bulbar) form of the disease, such data were not found in this study. The disease was equally distributed among sexes and the palpebral form was the most frequently encountered. Complications are uncommon. Other studies are needed to confirm the findings of this study. PMID- 14750227 TI - The use of a pinhole aperture during the recording of pattern reversal visual evoked potentials. AB - In young children, mentally retarded persons or in malingering persons, determining the optimal refraction is not always evident. Because of the importance of this optimal refraction during the recording of pattern-reversal visual evoked potentials, we tried in this study to overcome these refraction errors by the use of a pinhole. PMID- 14750228 TI - Alport syndrome and conjunctival telangiectasia. AB - We report a patient with ocular abnormalities due to Alport syndrome. Considerable improvement of visual acuity was obtained following ocular lens extraction with foldable lens implantation in the right eye to compensate the effect of the lenticonus anterior. The presence of conjunctival telangiectasia is a new clinical finding in a patient with Alport syndrome. PMID- 14750229 TI - Results of penetrating keratoplasty in syphilitic interstitial keratitis. AB - PURPOSE: To compare the results in our patient series after penetrating keratoplasty (PKP) for syphilitic interstitial keratitis (IK) with those described in the literature. METHODS: Retrospective case series in which visual acuity (VA), graft clarity, rejection episodes, intraocular pressure and endothelial cell density (ECD) were examined postoperatively. RESULTS: Postoperative VA improved in all cases. There was no evidence of wound dehiscense or occurrence of retrocorneal membrane formation in any case. Postoperative inflammation was not more severe in our patients with syphilitic IK compared to patients undergoing PKP for other reasons. A normal decline in ECD proved that there was no subclinical inflammation as well. CONCLUSION: PKP for syphilitic IK has a good prognosis in our case series as far as graft survival is concerned. Improvement in VA was present in all cases, though sometimes limited. In our case series, we experienced less postoperative complications than described in the older literature, which is probably due to better microsurgical techniques used nowadays. PMID- 14750230 TI - Gradenigo syndrome: a case-report. AB - We report a case of sixth nerve palsy as a rare complication of acute otitis media (apical petrositis). The clinical triad of acute otitis media, pain in the distribution of the fifth cranial nerve and sixth nerve palsy is known as Gradenigo syndrome. PMID- 14750231 TI - Ectopia lentis and central serous chorioretinopathy in a patient with a marfanoid habitus. A case report. AB - We report a case of ectopia lentis and central serous chorioretinopathy in a young female patient with several manifestations of Marfan syndrome in the skeletal, cardiovascular and dermatological systems. We can't conclude however that these two ocular abnormalities are linked. PMID- 14750232 TI - Lasek for myopia: first results. AB - PURPOSE: To evaluate the efficacy and safety of laser-assisted subepithelial keratectomy (LASEK) for the treatment of myopia and astigmatism (low, moderate and high). MATERIAL AND METHODS: Laser-assisted subepithelial keratectomy was performed by 3 surgeons in 45 eyes using an INPRO Gauss Excimer laser (31 eyes) or a NIDEK EC-5000 Excimer Laser (14 eyes). The mean follow-up is 9 months (3-15 months). Preoperative best-corrected visual acuity (BCVA) and corneal topography were measured. The postoperative parameters were: uncorrected visual acuity (UCVA), best-corrected visual acuity (BCVA), spherical equivalent refraction (SER), defocus equivalent refraction (DER), and corneal topography. RESULTS: The mean preoperative sphere and cylinder were -4.09 diopters (D) +/- 1.94 D (range 1.25 D to -9.75 D) and -0.67 D +/- 0.55 D (range 0 D to -2.50 D) respectively. No eye lost 2 or more lines of BCVA. The UCVA was > or = to 20/20 in 56% of the cases and 20/40 or better in 100% of the cases. No eye developed corneal haze that affected visual acuity. No major complications were recorded. CONCLUSIONS: LASEK treatment is a safe and effective technique for treatment of low to high myopia. This surgical technique is less invasive and more effective than LASIK because of the lack of flap- and microkeratome-related complications. PMID- 14750233 TI - Posterior uveitis: an under-recognized adverse effect of pamidronate: 2 case reports. AB - Ocular adverse effects of pamidronic acid are rare but well documented. Pamidronate, an inhibitor of bone resorption used primarily in the management of tumor-induced hypercalcemia and Paget's disease, is reported to cause conjunctivitis, anterior uveitis, and infrequently episcleritis and scleritis. It is hypothesized that an allergic or immunologic phenomenon caused by drug-indued immune complex formation is at fault. The reason why the uvea is a target organ is unclear. The acute inflammatory response seems unrelated to the dose of the drug, the way of administration, or the activity of Paget's disease or malignancy. We report two cases of pamidronate-induced posterior uveitis, following the WHO Causality Assessment Guide of Suspected Adverse Reactions. Uveitis and scleritis have been reported in association with a variety of topical, intraocular, periocular, and systemic medications. Seven criteria were proposed to establish causality of adverse events by drugs. Only systemically administered biphosphonates meet all seven criteria. Where pamidronate is currently considered as the drug of choice in diverse strategies, the adverse ocular effects should be well known to physicians in order to make rapid diagnosis and stop the drug causing adverse reaction. PMID- 14750234 TI - Patentable commodities? PMID- 14750235 TI - Nevus cell aggregates in lymph nodes. PMID- 14750236 TI - Immunohistochemical detection of Aspergillus species in pediatric tissue samples. AB - Definitive diagnosis of invasive aspergillosis often requires tissue samples for histologic evidence of fungal infection and culture confirmation of Aspergillus species. However, the culture frequently fails to isolate Aspergillus species. Alternative approaches to confirm Aspergillus infection use polymerase chain reaction, in situ hybridization, and immunohistochemical analysis on paraffin embedded sections. These approaches are well characterized in animals and adult patients but not pediatric patients. We studied the immunoreactivity of a commercially available monoclonal antibody, Mab-WF-AF-1 (DAKO, Carpinteria, CA), on paraffin-embedded sections from 16 pediatric cases with invasive aspergillosis, of which 12 were proven by culture. Optimal immunoreactivity required microwave antigen retrieval using high pH; 5 other antigen retrieval approaches were unsuccessful. With optimization, the monoclonal antibody was strongly immunoreactive in all cases with staining of the Aspergillus cell wall, septa, and cytoplasm. Background was minimal with no cross-reactivity to Candida albicans. These findings demonstrate the usefulness of the Mab-WF-AF-1 antibody in pediatric tissues suspected of invasive aspergillosis. PMID- 14750237 TI - Fatal West Nile virus encephalitis in a renal transplant recipient. AB - West Nile virus (WNV), a mosquito-transmitted single-stranded RNA flavivirus, causes human disease of variable severity. We report clinical and pathologic findings of fatal encephalitis from the transmission of WNV from an organ donor to a kidney transplant recipient. The patient developed a febrile illness 18 days after transplantation, which progressed to encephalitis. Postmortem examination demonstrated extensive viral encephalopathic changes. Immunohistochemical studies highlighted WNV antigens within neurons, especially in the cerebellum and brainstem. Flavivirus virions were detected ultrastructurally within the cerebellum, and WNV was isolated from the brain and the brainstem. Thus, this case demonstrates the first death in the first solid organ transplant-associated transmission of WNV. Immunosuppression of the transplant recipient might have been responsible for the fulminant viral effects. The pathologic diagnosis helped guide subsequent epidemiologic and laboratory studies. PMID- 14750238 TI - Expression of bcl-6 and CD10 protein is associated with longer overall survival and time to treatment failure in follicular lymphoma. AB - Follicular lymphomas (FLs) are a heterogeneous group of tumors, but prognostic factors are evaluated insufficiently in this common hematologic neoplasm. While bcl-6 and CD10 are expressed characteristically in FLs, their significance for biologic behavior of FL has not been studied previously. Samples from 73 patients with FL and clinical follow-up from 7 to 231 months were evaluated by immunohistochemical analysis. Patients with high levels of bcl-6 expression had favorable overall survival (OS) (P = .003), disease-specific survival (DSS) (P = .033), and time to treatment failure (P = .003) compared with patients with low levels of bcl-6 expression. Multivariate analysis showed that the results for OS, DSS, and time to treatment failure were independent of the international prognostic index. Patients with CD10+ FLs also had longer OS (P = .001), DSS (P = .007), and time to treatment failure (P = .004), and grade 1 FL was associated with better OS (P = .01) and a statistical trend for longer DSS (P = .05) and time to treatment failure (P = .05), but these results were not independent of bcl-6 expression or the international prognostic index in multivariate analysis. PMID- 14750239 TI - Eosinophil-rich CD30+ lymphoproliferative disorder of the oral mucosa. A form of "traumatic eosinophilic granuloma". AB - We describe 3 patients who had oral mucosal lesions with features of traumatic eosinophilic granuloma (TEG) and containing CD30+ atypical cells. In 1 patient, the oral lesion was followed by skin nodules. All lesions were evaluated histologically, by immunohistochemical analysis, and by polymerase chain reaction (PCR) analysis of the T-cell receptor (TCR) gamma chain gene. All oral lesions were characterized by a dense and deeply infiltrative lymphoproliferation, showing epitheliotropism and massive eosinophilia. They contained atypical large lymphoid cells, which expressed T-cell markers and CD30. PCR analysis showed a monoclonal rearrangement of the TCR gamma chain gene in all lesions and, in 1 patient, the same rearrangement in the oral and cutaneous specimens. The lesions in these patients seem to be the oral counterpart of the spectrum of primary cutaneous CD30+ lymphoproliferative disorders and should be recognized as such to avoid a diagnosis of large T-cell lymphoma and possible consequent overtreatment. However, they represent only a subset among several others within the complex and heterogeneous category of disorders referred to as TEG. PMID- 14750240 TI - Noninvasive monitoring of hemoglobin. The effects of WBC counts on measurement. AB - The efficacy of a noninvasive hemoglobin monitoring device (Astrin, Sysmex, Kobe, Japan) was evaluated for healthy volunteers and for patients with hematologic disorders. At the same time, the effects of WBC counts on noninvasive monitoring were studied by clinical evaluation and in ex vivo experiments. The hemoglobin levels determined by the device (Ast-Hb) and a conventional analyzer (T-Hb) were compared. The coefficient of correlation between findings with the Ast-Hb and the T-Hb for healthy volunteers was r = 0.626, whereas that for patients with hematologic disorders was r = 0.762. A comparison of the ratios of measurement errors in hemoglobin levels by Ast-Hb and T-Hb indicated that the number of WBCs had no effect on hemoglobin monitoring. Moreover, ex vivo studies using isolated WBCs and an optical model that imitates blood vessels and tissue in human fingers confirmed these results. Therefore, this new hemoglobin monitoring device can be expected to be useful for continuous hemoglobin monitoring. PMID- 14750241 TI - Nodal melanocytic nevi in sentinel lymph nodes. Correlation with melanoma associated cutaneous nevi. AB - Melanocytic nevi occurring in lymph nodes create diagnostic difficulty by mimicking metastases. Few studies describe nodal nevi in sentinel lymph nodes (SLNs) excised for melanoma. We evaluated 72 cases in which patients had undergone SLN biopsy for melanoma. Lymph nodes and cutaneous melanomas were evaluated according to a standard protocol. Nodal nevi were identified in 8 patients (11%). Of these, 6 (75%) had an associated cutaneous nevus (P = .006). Of 21 patients with an associated nevus, 4 (19%) with nodal nevi had a cutaneous nevus with congenital features (P = .01). The incidence of nodal nevus correlated with a Breslow thickness greater than 2.5 mm (P = .02). Nevi were not seen in non SLNs. Nodal nevi appear more frequently in patients with melanoma-associated cutaneous nevi, particularly if congenital features are present. The increased frequency of nodal nevi in SLNs relative to non-SLNs suggests an etiology of mechanical transport of nevus cells. PMID- 14750242 TI - Cost-effectiveness of immediate specimen adequacy assessment of thyroid fine needle aspirations. AB - Pathologists and cytotechnologists often provide immediate specimen adequacy evaluation of thyroid fine-needle aspirations (FNAs) to ensure that diagnostic material is obtained. We assessed the cost-effectiveness of this practice. All patients who had a thyroid FNA specimen accessioned at the Beth Israel Deaconess Medical Center, Boston, MA, during a 6-month period were included and divided into 2 groups: (1) with or (2) without immediate adequacy assessment. Specimen adequacy from each group was compared. The time spent to perform the adequacy assessment was recorded. Compared with group 2, group 1 had more specimens with diagnostic cellular material (67.2% vs 47.0%) and fewer specimens with suboptimal (23.3% vs 38.1%) or nondiagnostic cellular material (9.5% vs 14.9%) (P = .002). At the time of adequacy assessment, 98% (60/61) of the adequate specimens were obtained with 3 or fewer passes. The improved rate of diagnostic material was achieved at a cost of 220 minutes of cytologists' time per additional diagnostic specimen compared with group 2. It may be most cost-effective to routinely obtain 3 passes and to perform immediate adequacy assessment under special circumstances such as repeated procedures. PMID- 14750243 TI - Determination of HER-2 status and chromosome 17 polysomy in breast carcinomas comparing HercepTest and PathVysion FISH assay. AB - We evaluated and compared 2 HER-2 tests (immunohistochemical analysis [HercepTest, DAKO, Carpinteria, CA] and fluorescence in situ hybridization [FISH]) and assessed chromosome 17 polysomy status in relation to these tests. HER-2 status was obtained in 690 cases. The rinse step in the HercepTest before and after addition of the visualization reagent was 2 minutes in 188 cases and was increased to 5 minutes in 600 cases. HercepTest with both rinse steps was performed on duplicate slides in 98 cases. Chromosome 17 ploidy status based on FISH results was determined in 687 cases. Weak overexpression (2+) of HER-2 protein was not due to gene amplification in a majority of cases (67/76 [88%]). A small subset of breast carcinomas (19/687 [2.8%]) strongly overexpressed (3+) HER 2 protein without gene amplification. The aneuploidy rate was similar in negative and 2+ cases (60/141 [42.5%] and 12/26 [46%]), compared with 86% (18/21) in 3+ cases. The incidence of polysomy 17 in 2+ nonamplified cases (3/67 [4%]) was similar to that seen in negative cases (5.5%), in contrast with 47% (9/19) of 3+ nonamplified cases. Adding a longer rinse step to the HercepTest converted a subset (3/10 [30%]) of weakly positive cases to negative cases. Weak overexpression of HER-2 protein in a majority of cases seems to represent an artifactual staining pattern. Chromosome 17 polysomy is a major factor in strong HER-2 protein overexpression in 3+ nonamplified cases. PMID- 14750244 TI - COX-2, MMP-9, and Noguchi classification provide additional prognostic information about adenocarcinoma of the lung. A study of 117 patients from Brazil. AB - We report immunohistochemical staining results for cyclooxygenase (COX)-2 and matrix metalloproteinase (MMP)-9 in primary tumors of 117 patients with resected adenocarcinoma of the lung (median follow-up, 20 months). For COX-2, we graded the degree of tumor staining according to the sum of staining intensity and the proportion of cells staining. For MMP-9, we used morphometry to quantify cytoplasmic staining. We used the Cox proportional hazards model to analyze overall survival. With only 29 patients censored at last follow-up, after controlling for the effect of pathologic stage, staining for COX-2 and MMP-9 and subtype of tumor were related significantly to survival (P < 6 x 10(-5)). The effects of COX-2 and MMP-9 were opposite. Whereas any staining for COX-2 decreased the hazard and increased survival time, increased staining for MMP-9 increased the hazard and decreased survival time. The results also suggested that staining for COX-2 decreases with dedifferentiation. Our results suggest that staining for the combination of COX-2 and MMP-9 and categorizing tumors into papillary and nonpapillary types may provide important prognostic information for patients with resected adenocarcinoma of the lung; it is possible that these 3 variables could aid decisions about postoperative adjuvant treatment. PMID- 14750245 TI - Atypical glandular cells of undetermined significance. Outcome predictions based on human papillomavirus testing. AB - Cases of atypical glandular cells (AGC) diagnosed on liquid-based preparations were culled from a 3-year period. When available, residual cellular material was analyzed for human papillomavirus (HPV) by polymerase chain reaction and correlated with cytologic and histologic (biopsy) outcome. Of 178,994 cytologic cases, 187 (0.1045%) contained AGC compared with 8,740 (4.8828%) atypical squamous cells (ASC) for an AGC/ASC ratio of 0.021. HPV results and follow-up were available for 108 specimens from 106 patients. Depending on the end-point (histologic/cyto-logic), the sensitivity range of HPV testing for significant cervical disease (high-grade squamous intraepithelial lesion [SIL], adenocarcinoma in situ [ACIS], invasive carcinoma) was 83% with a specificity range of 78% to 82%, a positive predictive value of 57% to 61%, and a negative predictive value of 91% to 95%. Fifteen false-positive results included concurrent ASC or low-grade SIL, ASC on follow-up cytology, and previous ACIS with a negative follow-up cone biopsy result. Noncervical glandular neoplasia (including atypical endometrial hyperplasia) was confirmed in 13 cases (1 recurrent), only 2 of which scored positive for HPV. HPV-positive AGC has a substantially higher positive predictive value for significant disease than ASC (61% vs historic 20%) and merits consideration in the triage of patients with atypical endocervical cells not otherwise specified. However, noncervical or other HPV-negative glandular neoplasia must be considered in all patients with AGC, particularly older patients. PMID- 14750246 TI - Mesenteric fibromatosis with involvement of the gastrointestinal tract. A GIST simulator: a study of 25 cases. AB - Mesenteric fibromatosis (MF) and gastrointestinal stromal tumors (GISTs) are distinct lesions, but they often are confused with each other. Correct identification is essential because of their vastly different therapeutic and prognostic connotations. We reviewed 25 cases of MF with involvement of the wall of the gastrointestinal tract and found that GIST was the most common misdiagnosis (13 [52%]). MF was characterized by a spatially homogeneous proliferation of wavy spindle cells without atypia, associated with collagen deposition (often of the keloidal type), and an infiltrative border. Most cases displayed prominent muscular arteries and dilated, thin-walled veins. The mitotic count was relatively low, and no atypical mitotic figures were identified. These features are sufficiently characteristic of MF to permit distinction from GIST on the basis of routinely stained sections in the large majority of the cases, but immunohistochemical analysis provides a supporting role in the few equivocal cases. PMID- 14750247 TI - P504S immunostaining boosts diagnostic resolution of "suspicious" foci in prostatic needle biopsy specimens. AB - From 1.5% to 9.0% of prostatic needle biopsy specimens disclose atypical small acinar proliferations (ASAPs) suggestive of malignancy, carrying an approximate 45% predictive value for cancer. We applied keratin 34 beta E12 and P504S monoclonal immunostains to 93 cases that were judged as ASAP after H&E staining alone. Forty-one ASAP foci survived recutting for both immunostains. Three urologic pathologists independently assigned post-keratin 34 beta E12 diagnoses of cancer, ASAP, high-grade prostatic intraepithelial neoplasia, or benign and then reviewed P504S slides and assigned final diagnoses. Eight foci (20%) were resolved unanimously after keratin 34 beta E12 staining; 18 (44%) were resolved by 1 or 2 evaluators and 29 (71%) by at least 1. According to whether post keratin 34 beta E12 ASAP designation was given by 3, 2, or 1 evaluator(s), P504S immunostaining unanimously resolved an additional 5 (12%), 10 (24%), or 23 (56%) of 41 ASAP foci and cumulatively, 31 foci (76%). Among 35 men (excluding 6 with cancer in other cores of the original biopsy), these immunostains could have permitted cancer diagnosis in 11 (31%), without repeated biopsy. Thus, the consensus diagnosis rate improved from poor to good after supplementing 34 beta E12 immunostaining with P504S. PMID- 14750248 TI - Apoptosis of cardiomyocytes in explanted and transplanted hearts. Comparison of results from in situ TUNEL, ISEL, and ISOL reactions. AB - We assessed the efficiency of detecting myocyte apoptosis within human hearts using in situ enzymatic reactions in paraffin-embedded tissue samples: in situ end labeling (ISEL), terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling (TUNEL), and in situ oligoligation (ISOL). The reactions were carried out in explanted hearts (idiopathic dilatative cardiomyopathy, n = 6; ischemic heart disease, n = 3) and in endomyocardial biopsy specimens (EMBs; n = 32) obtained from transplanted human hearts. The results were verified by DNA laddering. The ISOL reaction led to a significantly (P = .027) smaller number of false-positive results (2/41 [5%]) compared with assessment by ISEL (9/41 [22%]) or TUNEL (9/41 [22%]). Only 1 ISEL+ apoptotic cardiomyocyte was found in specimens from explanted hearts. Among the EMBs, 1 specimens had TUNEL+ apoptotic cardiomyocytes and 1 specimen had ISEL+ apoptotic cardiomyocytes. This implies that verifying results by independent methods must be used for TUNEL and ISEL techniques. A smaller number of false-positive results makes interpretation of ISOL results easier, although the sensitivity of this reaction remains to be established. PMID- 14750249 TI - E-cadherin-negative primary small cell carcinoma of the breast. Report of a case and review of the literature. AB - Primary small cell carcinoma of the breast is exceedingly rare, with fewer than 25 reported cases. The case presented herein is that of a 61-year-old woman with a 2.5-cm mass of the left breast. She underwent mastectomy with axillary node dissection. Histologic examination revealed sheets and nests of small, hyperchromatic, malignant cells with indistinct nucleoli and scant cytoplasm. High-grade solid and comedo ductal carcinoma in situ also was present. Two of 5 axillary lymph nodes contained metastatic disease. Immunohistochemical analysis demonstrated weak immunoreactivity for cytokeratin, neuron-specific enolase, and bcl-2. This histologic and immunohistochemical profile was consistent with that of a primary small cell carcinoma. Interestingly, this neoplasm lacked immunoreactivity for E-cadherin. E-cadherin expression has been documented in all 11 (100%) of 11 previously reported cases of primary small cell carcinoma of the breast, suggesting that this tumor is a form of ductal carcinoma. To our knowledge, this is the first reported case of E-cadherin-negative small cell carcinoma of the breast, which raises the question of a possible lobular histogenesis in some of these neoplasms. PMID- 14750250 TI - Recombinant factor VIIa in the treatment of bleeding. AB - Recombinant factor VIIa (rFVIIa) has become available for treating people with hemophilia with inhibitors who experience bleeding or require surgery. It has become apparent that rFVIIa is useful in controlling bleeding in a variety of clinical situations. This review attempts to collate and summarize the nonhemophilia applications of rFVIIa. The theoretical mechanism for the coagulation and hemostatic effects of rFVIIa are discussed. The dosage and clinical administration are described. The potential uses for patients with liver disease, anticoagulation-induced bleeding, surgery, thrombocytopenia, thrombasthenia, von Willebrand disease, and other bleeding disorders are reviewed. The use of rFVIIa is evolving, and the indications, dosage, and precautions or contraindications need to be further described and defined. It is an expensive therapy and needs to be prescribed judiciously. This review is meant to be an introduction to this new hemostatic reagent. The uses for rFVIIa will evolve as more studies are published. PMID- 14750251 TI - Biphasic activated partial thromboplastin time waveform and adverse events in non intensive care unit patients. AB - The purpose of the present study was to determine whether the presence of a biphasic activated partial thromboplastin time (aPTT) waveform (BPW) is associated with adverse clinical outcomes among patients not in an intensive care unit (ICU). Consecutive patients from the emergency department or non-ICU inpatient floors with a BPW (n = 24) were enrolled prospectively, along with 24 matched control subjects with a normal aPTT waveform. Patients with a BPW had a significantly longer hospital stay (mean [median], 16.9 [11] vs 4.9 [2.5] days; P = .011), were more likely to have a positive microbial culture (16/24 [67%] vs 3/24 [13%]; P < .001), were transferred more often to an ICU (6/24 [25%] vs 0/24 [0%]; P = .010), and were more likely to receive an RBC transfusion (11/24 [46%] vs 5/24 [21%]; P = .047) or a fresh frozen plasma transfusion (5/24 [21%] vs 0/24 [0%]; P = .025). Emergency department patients with a BPW were more likely to be admitted (11/11 vs 5/11; P = .018). These results suggest that the BPW is associated with an increased rate of adverse events among non-ICU patients. Further study in this population is warranted. PMID- 14750253 TI - Peritoneal involvement and free tumor cells in peritoneal cavity of patients with colon cancer. PMID- 14750252 TI - Anti-beta 2-glycoprotein I and antiphosphatidylserine antibodies are predictors of arterial thrombosis in patients with antiphospholipid syndrome. AB - The predictive value (PV) and association of 4 antiphospholipid antibodies with clinical manifestations of the antiphospholipid syndrome (APS) were evaluated in 90 patients with systemic lupus erythematosus (SLE) and 100 with APS. Patients with APS were classified into arterial thrombosis, venous thrombosis, and pregnancy morbidity subgroups. IgG, IgM, and IgA anticardiolipin (aCL), antiphosphatidylserine (aPS), anti-beta 2-glycoprotein I (anti-B2GPI), and antiprothrombin (aPT) antibodies were determined by enzyme-linked immunosorbent assay. Individually, anti-B2GPI and aPS antibodies had the strongest PV for APS (86.4%-94.1%; P < .001) in patients with SLE. The PV for APS reached 100% when 2 or more antibodies were present. Similarly, anti-B2GPI and aPS antibodies had a stronger PV and association for arterial thrombosis (87%-95%; P < .001) compared with venous thrombosis (80%-92%; P = .01). Weak PV and association with pregnancy morbidity were seen with all antibodies. These results suggest an important pathogenic role of anti-B2GPI antibodies in arterial thrombosis. In addition, anti-B2GPI and aPS antibodies seem to provide the best diagnostic value for the laboratory assessment of APS. PMID- 14750254 TI - [Current methods in prevention and treatment of acute renal failure]. PMID- 14750255 TI - [Diabetic nephropathy in contemporary nephrology]. PMID- 14750256 TI - [Early detection of renal diseases-urgency or clever choice?]. AB - Economical end medical benefits of early referral and nephrological care were presented in patients with very early stages of renal disease. PMID- 14750257 TI - [Clinical aspects and diagnosis of Epstein-Barr virus infections]. PMID- 14750258 TI - [Standardization issues of immunoprocedures]. PMID- 14750259 TI - [Cytokines and bone turnover markers]. AB - OBJECTIVE: Bone turnover markers (osteocalcin, bone-ALP, beta-crosslaps--CTX) and cytokines (IL-1 alpha, IL-8 and IL-10) in hip joint fluid were analyzed in patients with aseptic loosening of prosthesis before revision surgery, in patients with rheumatoid arthritis and with idiopathic coxarthrosis for comparison. METHODS: Bone turnover markers were determined by electrochemiluminescence, colorimetric or ELISA method and cytokines by ELISA in joint fluid collected at the beginning of the surgery. RESULTS: Patients with loose implants had lower concentration of the resorption marker than cases with rheumatoid arthritis and coxarthrosis (7771 +/- 3322 vs 25986 +/- 16059 p < 0.01 and 23047 +/- 32556 pmol/L, p < 0.003) and over tenfold lower concentration of osteocalcin, the bone formation marker (p < 0.04 i p < 0.01). Concentration of IL 8 was elevated and similar in patients with loosening and rheumatoid arthritis while in cases with osteoarthrosis the mean value was twice lower. The anti inflammatory IL-10 was highly elevated only in cases with prosthesis loosening. Additionally, a negative correlation was observed between CTX and IL-10 was and positive between IL-10 and time to revision surgery. CONCLUSIONS: We conclude that increased local production of inflammatory cytokines leading to uncoupling of bone turnover is a part of the loosening process. PMID- 14750260 TI - [Hepatitis C virus infection--a major problem in hepatology]. PMID- 14750261 TI - [Peginterferon alpha and ribavirin therapy for difficult-to-treat chronic hepatitis C patients]. PMID- 14750262 TI - [Stem cells--future in transplantology]. PMID- 14750263 TI - [Chronic Chlamydia pneumonia--complications]. PMID- 14750264 TI - [Assessment methods of autoantibodies in autoimmune diseases]. AB - In most of the autoimmune diseases, a humoral and cellular immune response is characteristically seen, with autoantibodies and cells directed to distinct intracellular antigens. This phenomenon can be shown in systemic diseases like sclerosis, systemic lupus erythematosus, Sjogren's syndrome, mixed connective tissue disease, polymyositis and rheumatoid arthritis. It is also evident that autoantibodies are present in many the autoimmune diseases (called organ specific) like for example Hashimoto, Graves-Basedow or Addison disease. The presence of autoantibodies is important items to be considered in establishing a diagnosis and because of that autoantibodies are included in the diagnostic criteria of many autoimmune diseases. They are useful prognostic markers in some situations and facilitate clinical and treatment follow-up. Since year 1950 (discovering of the first autoantibody--classical rheumatoid factor IgM class) many new methods like: immunoelectrophoresis, counter-electrophoresis, double and radial immunodiffusion in gels, immunoagglutination and haemolytic methods have been used for autoantibodies assessment. It seems to us the indirect immunofluorescence method (IIF) was a most powerful, sensitive and comprehensive test for screening of autoantibodies, until an immunoenzymatic (EIA) methods (ELISA, Western-blotting) in late 60-s was worked out. The immunoenzymatic tests are very useful because of their simplicity and reliability. But there is one more excellent test hybrid named "Colorzyme" (presented by Immuno-Concept Corporation from USA) worked out by combining of the EIA and IIF tests. Instead of FITC-conjugates (like in IIF) a HRP-conjugates for developing of typical ANA test based on glass fixed Hep-2 cells have been used. The nuclear type of pattern we get using "Colorzyme" test are very strong, nit and rich in details. The prevalence of the "Colorzyme" test relies on that it can be properly done and interpreted by unexperienced technician. More and more new-founded resources of marker autoantibodies and methods force to introduction into standardization both methods and specimens on which they are marked. PMID- 14750265 TI - [Epidemiology of end stage renal disease in Poland]. AB - Development of end stage renal disease for many years was in Poland "signum maliominis" for patients with this syndrome. Situation dramatically changed as a consequence of dramatic development of renal replacement therapy. This progress was observed not only in basic forms of dialysis therapy (hemodialysis and peritoneal dialysis) but also renal transplantation. Increased availability of renal replacement therapy influenced both on the increment total number of treated patients (by 250% during period 1991-2001) and an qualification criteria. These changes caused substantial increase of patients with diabetic and hypertensive nephropathy. Such phenomenon is observed in whole the world and one can conclude that situation in this matter is normalizing in Poland. Similar situation is observed taking into account age of treated patients together with described above changes in primary diseases leading to end stage renal disease will cause more and more significant problem for health care system in Poland and many other countries. PMID- 14750266 TI - The social worlds of immigrant youth. AB - This introductory chapter uses a detailed case study to illustrate the interconnection of multiple social influences on one particular youth's path of migration. It further identifies some of the major influences on immigrant youth development, including the stresses of migration, separations and reunifications, changing networks of relations, poverty and segregation, and identity formation. PMID- 14750267 TI - Responsibilities of children in Latino immigrant homes. AB - Using survey and observational data, children's contributions to households in a Mexican immigrant community in Chicago are examined. Children provide essential help to their families, including translating, interpreting, and caring for siblings. These daily life activities shape possibilities for learning and development. PMID- 14750268 TI - Surfing the "model minority" wave of success: how the school context shapes distinct experiences among Vietnamese youth. AB - Vietnamese students must contend with the burden of the myth of being a model minority. As a result of adults' high expectations of them, Vietnamese youth receive structural and ideological advantages over other nonwhite racial groups. Further, the students themselves reinforce the model minority image as they attempt to attain educational mobility. The authors examine the role of two distinct school contexts within the same school that shape the academic outcomes of Vietnamese students contending with the pressures of being considered members of the model minority. PMID- 14750269 TI - Ethnic language schools and the development of supplementary education in the immigrant Chinese community in the United States. AB - This chapter seeks to unpack ethnicity through a close examination of ethnic language schools and the ethnic system of supplementary education in the immigrant Chinese community in the United States. It sheds light on the specific ways in which ethnic community organizations contribute to educational achievement. PMID- 14750270 TI - "He is everything": religion's role in the lives of immigrant youth. AB - Using emergent theme analysis of qualitative interview data in combination with quantitative survey data, the role of religion in the lives of immigrant youth was explored. Latino, Haitian, and Chinese teenagers described, in their own rich words, the significance of religion to them; their responses are reflected in themes that point to the potential protective role of religion for some immigrant groups. PMID- 14750271 TI - Gendered expectations and gendered experiences: immigrant students' adaptation in schools. AB - This chapter draws on longitudinal data to examine the role of gender in immigrant students' educational adaptation. Analyses show that over time girls receive higher grades and express higher future expectations than do boys. Compared with boys, immigrant girls are more likely to be protected from risk factors, such as harsh school environments, by a supported network of teachers, friends, and parents, and to benefit from the shield of ethnicity more than their male counterparts in their pursuit of education. PMID- 14750272 TI - Media in the lives of immigrant youth. AB - This chapter uses data from the Longitudinal Immigrant Student Adaptation Study to examine media use patterns among immigrant teens. Similarities and differences in media ownership levels, use patterns, and content preferences between immigrant teens and the U.S. teen population as a whole, as well as across immigrant groups and gender, are explored. Implications for educators and others who work with immigrant youth include active discussion with youth about their media choices and media messages. PMID- 14750273 TI - [Post-traumatic stress disorder (PTSD). Assessment of its rate of occurrence in the adult population of Portugal]. AB - AIM: This is the first epidemiological study of posttraumatic stress disorder (PTSD) within the adult population (over 18 years of age) in Portugal. This study is particularly relevant given the proportion of the population exposed to trauma: 800,000 men participated in 14 years (from 1961 to 1975) of Portugal's colonial wars. METHODS: A representative sample of the adult nation-wide population was defined using bi-phasic and stratified methodology. In accordance with study inclusion/exclusion criteria, subjects from all geographical regions were selected, either randomly from the community or via the protocol defined stratification. Subjects identified as having been exposed to one or more traumatic events were further interviewed and assessed with the Short Screening Scale (Breslau, 1999) to determine the presence/absence of DSM-IV defined PTSD. FINDINGS: A representative sample of 2,606 subjects (53.3% men) was selected, aged 18 to 99 years old (mean = 43 years). Overall PTSD prevalence rate was 7.87% (205/2,606 subjects) with a statistically significative different distribution by gender: 4.8% for male and 11.4% for female subjects. Around 75% of the Portuguese adult population was exposed to at least one traumatic situation and 43.5% to more than one. We found that 11.6% of the sample were male exposed to combat/war and 9.9% of those developed PTSD. The most referred traumatic situation was Sudden violent death of family or friend, (29.3%), followed by Robed or assaulted, (22.7%) and Witnessing of serious accident or death, (22.2%). CONCLUSIONS: The prevalence of posttraumatic stress disorder (overall and by gender) was similar to that reported for other developed countries; lower than expected, given the extent of the population's exposure to combat. PMID- 14750274 TI - [Assessment of severity of meningococcal sepsis in children]. AB - Despite advances in critical care medicine, acute meningococcal infection remains complicated by high mortality. Different prognostic scoring systems have been developed but none of them is largely used. The objective of this study was to evaluate the performance at admission to the pediatric intensive care unit (PICU) of five severity scores in children with proven and unproven meningococcal infection. Our results seem to indicate that the Neisseria Sepsis Index (NESI) and the Rotterdam Score (RS) perform better than the other scores, being appropriate tools to assess severity of illness at admission to the PICU in children with proven or presumed meningococcal infection. PMID- 14750275 TI - [Pregnancy and the thyroid]. AB - The author points out the changes occurred in the axis hypothalamus/hypofysis/thyroid during normal pregnancy. They mention fetus thyroid development different stages, as well as the thyroid hormone secretion through the fetus. They also give special attention to those pregnancies and its special features occurring in places where the amount of iodine isn't enough and how treatment must be carried out. PMID- 14750276 TI - [Hypothyroidism and pregnancy]. AB - The authors refer hypothyroidism epidemiology and physiology, enhancing the need to raise thyroid hormones and also to adjust the doses of levothyrosine, normally used in the treatment. They describe the obstetric complications that can occur in non medicated hypothyroidism pregnancies, as well to the new-born baby. PMID- 14750277 TI - [Hyperthyroidism and pregnancy]. AB - The authors describe most common hyperthyroidism aetiologies that occur during pregnancy and also the immunity changes that define Graves' disease and the immunity tolerance associated to pregnancy. They also refer the clinical picture, laboratorial diagnosis and the treatment to be carried out. Medical treatment is mostly based on anti-thyroid drugs. Their characteristics and use during pregnancy are discussed. PMID- 14750278 TI - [Hyperemesis gravidarum and hyperthyroidism]. AB - The author describes hyperemesis gravidarum clinical, biochemical and hormonal chart. He also refers the features of HCG molecule, its connections with hormones, which there are also glycoprotein and its plasmatic changes in normal pregnancy. The association between hyperemesis gravidarum and hyperthyroidism is extremely important in the patient's clinical evaluation. He also alludes to the special hyperthyroidism features concerning hyperemesis gravidarum, as well as when and how to treat it. PMID- 14750279 TI - [Goiter and pregnancy]. AB - The author refers to the changes occurred in thyroid dimensions during pregnancy, evaluated clinically and echographically. He also describes the changes in thyroid normal function on pregnant women and the different stages of the fetal thyroid maturation. Some complementary tests must be carried out, when there is a presence of a goitre nodule, which opportunity and relevance the need are discussed. The possibility of thyroid carcinoma is referred as its most adequate treatment. PMID- 14750280 TI - [Thyroid nodule and pregnancy]. AB - The author refer that thyroid nodule occur quite often in general population. He also mention research methods, and how they could be changed when pregnant women in concern. Therefore this prognosis must be done through cytology and ultrasounds. When related to rate of occurrence, metastasis and mortality, there are no differences between pregnant or non-pregnant women with thyroid papillary carcinoma. He also discusses the options and when it is the right time to surgery. PMID- 14750281 TI - [Thyroiditis and pregnancy]. AB - Authors develop the concept of immunity tolerance and its importance in thyroiditis occurrence and evolution during pregnancy as well post-partum pathology. They also refer the mechanisms that can induce to change the immunity response during pregnancy involving maternal, placental and fetal cytochemistry. It is also proposed a constant and precocious screening of the thyroid dysfunction in pregnant woman with thyroid antibodies, because their positivity prevalence is extremely high as far as the associated risk of abortion. PMID- 14750282 TI - [Thyroid carcinoma and pregnancy]. AB - The authors refer particular aetiological features, prognostic e different types of thyroid carcinoma treatment on pregnant women. There are also other kind of tumours occurring during this time, although not so frequent. There are some protocols in discussion to treat these kind of thyroid carcinoma, considering that the problem must be specified according to the type of tumour, stadium, pregnancy time and the patient psychological state. PMID- 14750283 TI - [Thyroid dysfunction in the newborn]. AB - The authors refer to congenic hypothyroidism, hypotyroxinemia in premature new borns, thyroid dysfunction, congenic thyrotoxicosis, breastfeeding, composed anti thyroid drugs and children nutrition based on iodine. Concerning to the fetus and new-born thyroid dysfunction, it's very important to make an early diagnosis in order to avoid later complications. Several studies revealed that the different types of composed anti-thyroid drugs can induce pregnant woman choice. PMID- 14750284 TI - [Postpartum thyroiditis]. AB - The author refers to the main clinical symptoms, diagnosis, treatment and evaluation of the post-partum thyroiditis. Researches can reveal different post partum thyroiditis, considering the diagnosis and screening. Normally the classical presentation of post-partum thyroiditis includes a period of thyrotoxicosis, followed by hypothyroidism and finally thyroid functions in its normal levels. According to some statistic investigation, authors mention that after a period of normal and stable status, hypothyroidism can prevail as a long term disease. PMID- 14750285 TI - [Other postpartum thyroid dysfunctions]. AB - The author refers Graves' disease as the second most common cause of post-partum thyroid dysfunction. It is more common a long term persistence of hyperthyroidism but a transitory hyperthyroidism can be also possible. The disease hypothalamus/hypophysis can also be responsible for the post-partum thyroid dysfunction, even though it is extremely rare. The auto-immunity of hypophysis plus Sheehan syndrome are two main reasons for hypothyroidism, associated with other hypophysis hormones production deficit. PMID- 14750286 TI - [Pregnancy and breastfeeding. Side effects of drugs used in the treatment of thyroid diseases, on the fetus and the newborn]. AB - The authors allude to the composed anti-thyroid drugs, blocking drugs, iodides, radioactive iodine, lithium carbonate and tyrosine side effects. The most common are composed anti-thyroid drugs and tyrosine. Anti-thyroid drugs risks are related to the tresspassing of the placenta barrier which can induce in goitre and hypothyroidism. Thyroid hormones are also very important for the fetus neural development during the first quarter when they cross the placenta. PMID- 14750287 TI - [Uterine tumor resembling ovarian sex-cord tumor]. AB - Uterine tumors resembling ovarian sex-cord tumors are rare and few cases are published in the literature. The authors describe a recently diagnosed one. The clinical features consisted of: abnormal vaginal bleeding and enlargement of the uterus, which allowed confusion with several types of uterine pathology: benign or malignant one; the diagnosis is exclusively histopathological. These tumors have a benign biological behaviour. PMID- 14750288 TI - [Acral skin necrosis in a patient with malaria]. AB - A case of acral skin necrosis in a 21-year-old farm workman, associated with Plasmodium falciparum malaria is reported. Patient presented jaundice, disorientation and delirium, high parasitemia, hypoglycemia, anemia, low platelets, and dark necrotic maculae surrounded by an erythematous halo, circumscribed to toes. After malaria treatment, the clinical evolution was uneventful and the skin lesions also improved without sequels. The possibility that some of the skin changes, eventually observed in malaria patients, may be related with responses to Plasmodium infection is emphasized. PMID- 14750289 TI - Cost-effectiveness analysis of the New South Wales adult drug court program. AB - In New South Wales, Australia, a cost-effectiveness evaluation was conducted of an adult drug court (ADC) program as an alternative to jail for criminal offenders addicted to illicit drugs. This article describes the program, the cost effectiveness analysis, and the results. The results of this study reveal that, for the 23-month period of the evaluation, the ADC was as cost-effective as were conventional sanctions in delaying the time to the first offense and more cost effective in reducing the frequency of offending for those outcome measures selected. Although the evaluation was conducted using the traditional steps of a cost-effectiveness analysis, because of the complexity of the program and data limitations it was not always possible to adhere to textbook procedures. As such, each step involved in undertaking the cost-effectiveness analysis is discussed, highlighting the key issues faced in the evaluation. PMID- 14750290 TI - An evaluation of collaborative interventions to improve chronic illness care. Framework and study design. AB - The author's dual-purpose evaluation assesses the effectiveness of formal collaboratives in stimulating organizational changes to improve chronic illness care (the chronic care model or CCM). Intervention and comparison sites are compared before and after introduction of the CCM. Multiple data sources are used to measure the degree of implementation, patient-level processes and outcomes, and organizational and team factors associated with success. Despite challenges in timely recruitment of sites and patients, data collection on 37 participating organizations, 22 control sites, and more than 4,000 patients with diabetes, congestive heart failure, asthma, or depression is nearing completion. When analyzed, these data will shed new light on the effectiveness of collaborative improvement methods and the CCM. PMID- 14750291 TI - Characteristics of adolescents who provide neither parental consent nor refusal for participation in school-based survey research. AB - Schools offer a convenient setting for research on adolescents. However, obtaining active written parental consent is difficult. In a 6th-grade smoking study, students were recruited with two consent procedures: active consent (parents must provide written consent for their children to participate) and implied consent (children may participate unless their parents provide written refusal). Of 4,427 invited students, 3,358 (76%) provided active parental consent, 420 (9%) provided active parental refusal, and 649 (15%) provided implied consent (parental nonresponse). The implied consent procedure recruited more boys, African Americans, students with poor grades, and smokers. This dual consent procedure is useful for collecting some limited data from students who do not provide active consent or refusal. PMID- 14750292 TI - A comparison of methods to obtain active parental consent for an international student survey. AB - Many school-based research efforts require active parental consent for student participation. Maximizing rates of consent form return and agreement is an important issue, because sample representativeness may be compromised when these rates are low. This article compares two methods for obtaining active parental consent: return of consent forms in the mail versus return by students to their classrooms. The methods were tested in a pilot study of 46 schools (1,058 students), with half of the schools randomly allocated to each of the alternative methods. A hierarchical nonlinear model of consent form return and agreement rates suggests that the student-delivered method is more successful at producing higher rates of consent form return and agreement to participate in the study, after controlling for school-level characteristics. The authors discuss the findings and their implications for other researchers engaged in school-based research with adolescents. PMID- 14750293 TI - At least say thanks. PMID- 14750294 TI - An open question. AB - The lungs are surrounded by the pleural membranes. The visceral pleura directly covers the lung and is separated from the parietal pleura by a layer of surfactant, which reduces friction during respiratory movement. A potential space exists between these two layers, and they may become separated by fluid or air. A lung can collapse to the size of a fist under pressure from either. Standard treatment in the field for an open chest wound is an occlusive dressing. The first thing that can be used to occlude the wound is a gloved hand. After placing the dressing, evaluate the breath sounds and determine if they have improved. The dressing should be taped down on three sides, leaving one side open to relieve the pressure during exhalation (one-way valve). "Burping" the dressing involves lifting one side to make sure any pressure buildup is relieved, as occasionally the dressing can become adhered to the skin, which may lead to a tension pneumothorax. If, after ensuring the occlusive dressing is properly in place, the respiratory rate increases, distress level worsens, oxygen saturations fall and breath sounds decrease, then needle decompression is required. A neurovascular bundle is located underneath each rib, and it is important to avoid damage to that bundle by performing a decompression over the top of a rib. If the patient is intubated before the development of a tension pneumothorax, carefully evaluate the breath sounds (especially if the left-side sounds are diminished) to determine if the ET tube needs to be withdrawn a centimeter. The rescuer performing ventilation will usually recognize a tension pneumothorax by the difficulty in bagging the patient. Remember, when you perform a needle thoracentesis, you are creating an open chest wound. Early signs and symptoms of a tension pneumothorax include diminished or absent breath sounds, severe dyspnea, narrowing pulse pressure, tachycardia and restlessness. Neck veins may be distended, but this can be a normal finding in a supine patient. The classic sign is a deviated trachea; the trachea shifts toward the "good" lung as the buildup of pressure collapses the "bad" lung. This is a late sign and suggests the tension pneumothorax has been developing for some time. One sign that does not normally accompany a plain pneumothorax is hypotension. In this case, the persistent low BP, combined with cool, mottled skin and a delayed capillary refill time, led providers to suspect that a hemothorax was developing as well. With endotracheal intubation and pleural decompression, the positive-pressure ventilations allowed the affected right lung to inflate more fully, utilize more of the available alveolar space and "bag out" some of the blood pooling at the base. The patient's vital signs and saturation improved. He needed surgical treatment and removal of the blood in the pleural space before ventilation and oxygenation could normalize. PMID- 14750295 TI - Operating in orange: what do you do? PMID- 14750296 TI - Fireground EMS. AB - The incident commander must ensure that EMS is able to effectively communicate and function as the incident warrants. EMS should provide appropriate services to the general public and emergency responders. With routine cross-training and drills, EMS functions can become common practice when the incident is anything but routine. PMID- 14750297 TI - Clearing the airway. PMID- 14750298 TI - Three cardinal sins in airway management. PMID- 14750299 TI - Pediatric pulmonary emergencies. PMID- 14750300 TI - Not so basic airway maneuvers. PMID- 14750301 TI - Ten budgetary best practices. Staying out of the red in EMS. PMID- 14750302 TI - Documenting for dollars. PMID- 14750304 TI - Are you failing to communicate? PMID- 14750303 TI - When they just say no. Documenting patient refusals in the era of HIPAA. PMID- 14750305 TI - Practitioners in pediatric rehabilitation have endorsed the concepts of family centered service as an approach to the delivery of services to children with disabilities and their families. PMID- 14750306 TI - Parental adjustment to having a child with cerebral palsy and participation in home therapy programs. AB - Based on a qualitative research project guided by grounded theory methodology, this article describes parents' participation in home therapy programs recommended by physical and occupational therapists. The study involved in-depth interviews with 8 parents and 4 therapists of children with cerebral palsy. The parents reported that in the initial stages of adjustment to the diagnosis, they were typically overwhelmed by their situation and unable to participate in providing activities at home for their child. Furthermore, they concealed their lack of participation from the therapists, with whom they have not yet developed a trusting relationship. Although this study was conducted in New Zealand with a small number of parents, the findings raise questions about parent participation in the early days of therapy. It is suggested that service delivery needs to be organised in a way that will allow therapists to provide higher levels of input in the early stages while parents come to grips with what is required of them. The need for further research to determine the ways in which the therapy program or the therapist could facilitate parental participation, and how therapists might evaluate parents' readiness to participate in a home program is recommended. PMID- 14750307 TI - Use of the Alberta Infant Motor Scale to characterize the motor development of infants born preterm at eight months corrected age. AB - The Alberta Infant Motor Scale (AIMS) was used to examine variations in motor development of infants born preterm. Sixty infants attending a Developmental Follow-up Clinic participated. Infants were assessed by physical therapists using the AIMS and independently judged by physicians to be neurodevelopmentally and neurologically "normal," "suspect," or "abnormal." The AIMS clearly differentiated infants in these three categories. Compared to the normative sample, infants judged to be "normal" demonstrated similar motor behaviors, infants judged to be "abnormal" were significantly different across a wide range of items, and infants judged to be "suspect" were significantly different on items requiring antigravity postural control, lower extremity dissociation, and trunk rotation. The AIMS can be used to identify infants developing abnormally, to affirm normalcy in infants developing typically, and to identify motor differences in infants who are neurologically "suspect." In the latter group of infants, the AIMS can be used to provide anticipatory guidance to parents regarding the components of movement they might expect their infants to be developing next. PMID- 14750308 TI - The relationship between postural control and fine manual dexterity. AB - The influence of the development of postural control of the trunk and center of the body on the development of fine manual dexterity ability is a common assumption. The objectives of this study were: (1) to examine the relationship between the adult version of the Nine-Hole Peg Test and the "children's version" in order to establish construct validity for the latter, and (2) to investigate the relationship between postural control and fine motor performance of children developing typically aged five to six. The study sample included 47 children, 25 males and 22 females, with a mean age of 5 years and 8 months and a standard deviation of 3.8 months. Two tests were administered: (1) The balance subtest of the Bruininks-Oseretsky Test of Motor Proficiency (BOTMP; Bruininks, 1978); and (2) the Nine-Hole Peg Test (NHPT; Kellor, Frost, Silberg Iversen & Cumming, 1971), as well as the "children's version" of rotation and translation tasks (Case-Smith, 1993, 1995). The adult version of the NHPT revealed significant moderate correlations with most of the BOTMP tasks for both hands. Secondary results of the study support the concurrent validity of the two "children's version" tasks. Results show low to moderate correlations between performance on the NHPT subtests and the BOTMP subtest tasks, with significant correlations ranging from -.31 to -.47. Given the negative and low correlations obtained in this study, it is suggested that more extensive longitudinal research examine the relationship between postural control and fine motor manual dexterity performance. PMID- 14750309 TI - Social participation for children with developmental coordination disorder: conceptual, evaluation and intervention considerations. AB - Children with Developmental Coordination Disorder (DCD) often have limited participation in social activities, causing isolation, anxiety, emotional and social problems in children themselves and their families. This paper reviews studies that investigated participation in activities of children with DCD in home, community and school. Assessments to evaluate social participation of children with DCD, intervention, strategies and future research considerations for children with DCD are suggested. PMID- 14750310 TI - Predicting length of service provision in school-based occupational therapy. AB - Parents and school administrators commonly ask how long a child is likely to need school-based occupational therapy services. In an effort to provide a statistically supported answer, a record review of 464 cases of students discharged from school-based occupational therapy services in the past 7 years was performed. This review demonstrated that some general statements about service patterns could be determined for this particular caseload. Of note was the significant variability in length of services for students with multiple disabilities as compared to the more discrete length of service provision for students with speech impairments, learning disabilities and those students without a specific classification. The applicability to other practice settings is limited, but this study serves as a beginning point for discussion on the topic. PMID- 14750311 TI - Caught in the Web. PMID- 14750312 TI - [Drug resistance mediated by ABC transporters]. AB - Remarkable advances have been made in cancer chemotherapy by developing new anticancer drugs and pharmacogenomics strategies. However, multidrug resistance in human cancers is the major obstacle to long-term, sustained patient response to chemotherapy. Several ATP-binding cassette (ABC) transporters cause multidrug resistance in cancer cells by actively extruding the clinically administered chemotherapeutic drugs. P-glycoprotein (ABCB1/MDR1/P-gp) and MRP1 (ABCC1/GS-X pump) have been well characterized in terms of their molecular structure and function. In addition, ABCG2/breast cancer resistance protein (BCRP) is the most recently identified/ABC transporter, and is also reportedly associated with cellular resistance against chemotherapeutic agents, such as DNA topoisomerase I, II inhibitor. It is important to note that these ABC transporters are expressed not only in cancer cells but also in normal tissues to play a pivotal role in the absorption, distribution, and excretion of endogenous substances as well as xenobiotics. ABC transporters are key factors that can affect the pharmacokinetic profiles of drugs. Recent studies have revealed that many single nucleotide polymorphisms (SNPs) reside in these ABC transporter genes. Functional analysis of the genetic polymorphism of ABC transporters would greatly contribute to our understanding of individual differences in the drug response and also to the development of personalized medicine in the near future. PMID- 14750313 TI - [Role of oncologic nurses in cancer chemotherapy]. AB - The field of cancer chemotherapy requires specialized knowledge and skills, and nurses working in this field undergo specialized training. Oncologic nurses have a large role in cancer chemotherapy and demonstrate superior practical ability while also serving as counselors to patients and as promoters and coordinators of team-based medical care. They also work to carry out education and research on nursing in the field of cancer chemotherapy. PMID- 14750314 TI - [Management of pharmacists on the cancer chemotherapy]. AB - The duties of hospital pharmacists include drug dispensing, drug management, preparation of injections, drug information provision, in-hospital preparation, drug management guidance (centering on patient compliance instructions), tests and research, and education. The Department of Pharmacy conducts most of these activities, which are not noticed by other medical staff. As members of medical care teams, pharmacists must share information and perform their jobs from a pharmaceutical viewpoint in order to fulfill their duties as drug specialists, which include management of patient conditions, patient-oriented patient compliance instructions, risk management, management of dosages and administration methods for anticancer drugs during cancer chemotherapy, discovery of the initial symptoms of adverse drug reactions and prevention of adverse drug reactions to obtain the optimum effect, education on self-care after discharge from the hospital, and evaluation of drug economics for cancer chemotherapy. It is important that they acquire sophisticated expertise and stay in as close contact as possible with patients and their families. PMID- 14750315 TI - [Roles of clinical psychologists in cancer chemotherapy]. AB - Cancer patients face a variety of problems and undergo psychological turbulence during the clinical course of their disease. In particular, problems related to treatment, such as chemotherapy, can pose a great burden and place the patients in an unstable status, involving both anxiety and expectation related to problems faced in daily life and the unreliability of the therapeutic effects. To cope with the various psychological problems experienced by cancer patients, individual counseling, group therapy, and behavioral therapy, including progressive muscle relaxation training and autonomic training, are employed, depending on the situation of each individual patient. Clinical psychologists participating in cancer treatment need a basic knowledge and understanding of the pathological characteristics of cancer, methods of treatment, and the characteristic clinical course of various cancers, in order to be in close contact and maintain communication with physicians, nurses, and other medical staff, and to provide the support demanded by the patients and their family through a variety of approaches. PMID- 14750316 TI - [Chemotherapy and clinical research coordinator]. AB - We have described the activities and responsiveness of clinical research coordinators (CRC) who support clinical studies of cancer chemotherapy. Clinical studies are essential for approval of new chemotherapeutic drugs. To perform clinical studies with high quality control, CRC staffs should play a large coordinating role throughout the process of each study. We introduce the recent status of the support system for clinical studies and discuss its problems. PMID- 14750317 TI - [Cancer chemotherapy and electronic medical record]. AB - In the clinical setting, the functions required for electronic medical records for chemotherapy are proposed from a system developed for individuals. The directivity of a future electronic medical record system is considered in the present condition of an electronic medical record system and a medical system expansionist policy from the history of medical information from Japan. PMID- 14750318 TI - [Participation of pharmacy in the clinical path for cancer chemotherapy]. AB - The clinical path is about standardizing the clinical progress of a patient and his or her medical treatment. It is the technique of charting the order and timing of intervention of the physician, nurse and all other medical staff, and performing and evaluating them. The clinical path is also applicable to chemotherapy. We created two items: A "medicine management check sheet," which is the check list used by pharmacists for each medicine or therapeutic class, and a "medication counseling and instruction sheet," which standardizes the instruction of medicine management. In adopting the clinical path, we brought these two items together, calling it "the worksheet for pharmacists." It is utilized for medication instruction. Furthermore, we made adjustments to each department in our role as Clinical Path Committee secretariat. We collected guidelines and a database about the diseases adopted to the clinical path and utilized them for evaluation of the clinical paths. PMID- 14750319 TI - [Clinical study of the area under the blood concentration-time curve of targeting intra-arterial infusion chemotherapy with nedaplatin for primary oral cancer]. AB - We treated oral cancer using a targeting intra-arterial infusion chemotherapy with CDGP (nedaplatin); dosages were determined by the original formula. We compared the expected AUC (area under the curve) with the actual AUC in primary oral cancer cases to assess the optimal dosage of CDGP for intra-arterial chemotherapy and to study relevance of AUC, effectiveness of independent chemotherapy and side effects. Eleven cases were treated at the Department of Oral and Maxillofacial Surgery, Faculty of Medicine, Kagawa Medical University, from October 1998 to June 2002. The results are as follows. 1) A correlation was seen between the AUC and degree of thrombocytopenia. 2) AUC may reveal renal function, especially for GFR. 3) The response rate of the targeted intra-arterial infusion chemotherapy with CDGP was remarkably high and serious side effects were not observed. It appears that the set-up of the optimum dose should be decided based on the actual AUC. 4) There was about a 4/3 difference between the value of the expected AUC and the actual AUC. The equation for dosages based on the actual AUC needs to be studied further. PMID- 14750320 TI - [Neoadjuvant chemoradiotherapy for advanced squamous cell carcinoma of thoracic esophagus]. AB - Advanced thoracic esophageal cancer has a poor prognosis despite advances in surgery, such as three-field lymph node dissection. Multimodal therapy is needed to improve local control, resectability and survival rate. Fifteen patients with advanced squamous cell carcinoma of the thoracic esophagus were treated with neoadjuvant chemoradiotherapy (NAC) combined with concurrent radiation (30 Gy/12 f) and 3 courses of 5-FU and CDDP (CDDP 5 mg/m2/day + 5-FU 250 mg/m2/day: day 1 5: div). In the absence of unresectable disease and surgical risk, 12 patients underwent esophagectomy (Group 1) and 3 patients underwent additional chemoradiotherapy because of high surgical risk (Group 2). Side effects consisted of nausea, vomiting and myelo-suppression in 8 patients, but all patients tolerated and completed a full course of NAC. The effective rate (CR + PR) of NAC was 58.3% in Group 1 and 66.7% in Group 2. No patients showed pathological CR. Two-year survival rate was 28.1% in Group 1 (PR: 33.3%, NC: 20.0%) and 33.5% in Group 2. This protocol had acceptable toxicities but did not show survival benefit. Further trials are necessary to improve survival rate. PMID- 14750321 TI - [Phase I study of gemcitabine (GEM) and UFT combination chemotherapy for unresectable/recurrent pancreatic cancer]. AB - A phase I study of gemcitabine (GEM) and UFT combination chemotherapy was performed for unresectable pancreatic cancer. The regimen included UFT orally from day 1 through day 6 and from day 8 through 13, GEM intravenously on day 7 and 14, and 1 week rest as 1 course. Administration of more than 2 courses was attempted for each patient, with the same dosage levels at all 3 scheduled steps. Nine patients were enrolled in the study. Leukopenia and neutropenia of grade 3 occurred in 2 of 6 patients at level 2, and maximum tolerated dose (MTD) was considered to be 800 mg/m2 for GEM and 250 mg/m2 for UFT. Clinical effects of the patients included: 2 PR, 4 NC, and 3 PD, for a response rate of 22% (2/9 patients). Reduced CA19-9 level of less than half of that at starting time was observed in 4 of 9 (44%) patients. The recommended dose for the phase II study is GEM 800 mg/m2 and UFT 250 mg/m2 with consideration of application to outpatients and continuing courses. PMID- 14750322 TI - [Appropriate duration of postoperative oral adjuvant chemotherapy with HCFU for colorectal cancer]. AB - We conducted a joint study of different duration of drug administration for oral adjuvant chemotherapy using camphor (HCFU) with patients having advanced colorectal cancer who underwent curative resection. The patients were randomly divided into 2 groups, according to length of HCFU administration (6-month group and 2-year group), and followed up for 5 years postoperatively. In total, 239 patients were originally enrolled, out of which 155 were chosen as subjects for this study. There was significant difference in the overall cumulative 5-year survival rate between the short-term group and the long-term group (78.1% vs 89.6%). Between the respective subgroup that was defined by tumor location (colon or rectum), no differences were observed, but there was significant difference in the subgroup that was defined by the presence/absence of lymph node metastasis (59.4% vs 83.9%). It appears that oral adjuvant chemotherapy with HCFU is more effective when administered for 2 years than for 6 months. PMID- 14750324 TI - [Neurotoxicity of weekly TP (weekly paclitaxel + consecutive low-dose CDDP) therapy]. AB - OBJECTIVES: To evaluate drug-induced neurotoxicity with combination consecutive low-dose CDDP (CLD-CDDP) and weekly paclitaxel in comparison with TJ therapy. Neurotoxicity was judged according to accumulative scores on questionnaires. SUBJECTS AND METHODS: Weekly TP therapy (CDDP: 10 mg/m2, days 1-7, paclitaxel: 60 80 mg/m2, days 1, 8, 15) and TJ therapy (paclitaxel: 170-180 mg/m2, 3 hours, carboplatin AUC 4-5) were given to 24 subjects in 57 courses and 19 subjects in 30 courses as previously reported, respectively. RESULTS: 1. Significant differences were found in numbness accumulative scores (p = 0.011) between weekly TP and TJ therapy, but not in pain accumulative scores (p = 0.926) between weekly TP and TJ therapy. 2. Peak pain at days 3-4 of paclitaxel that appeared during TJ therapy disappeared during weekly TP therapy, while numbness persisted to a lesser extent with weekly TP therapy than with TJ therapy. 3. Numbness was observed to accumulate in some subjects but not in others during weekly TP therapy. CONCLUSION: The patients' subjective symptoms were able to be objectively evaluated with the questionnaires. Weekly paclitaxel was observed to reduce neurotoxicity, which might be reinforced in combination with CDDP. PMID- 14750323 TI - [Clinical evaluation of cefozopran as treatment for febrile neutropenia]. AB - Clinical effects and safety of cefozopran (CZOP) were evaluated by the Okayama Bone Marrow Transplantation Group. Twenty-five patients expected to experience febrile neutropenia during induction chemotherapy or consolidation chemotherapy of acute leukemia were enrolled between July 2000 and November 2002. CZOP was administrated by drip infusion at 4g/day bid for a minimum of 3 days. The clinical effects and safety were evaluated in 20 patients with fever of 37.5 degrees C or more from a clinically suspected infection. The underlying disease was acute myeloid leukemia in 17 patients, acute lymphoid leukemia in 1 and acute promyelogeneous leukemia in 1. The complicating infections were sepsis and suspected sepsis. Clinical efficacy was excellent in 11 patients, good in 1, fair in 2 and poor in 6, with an efficacy rate of 60.0%. The efficacy rate in patients whose albumin levels before therapy were less than 3.8 g/dl was 37.5%, whereas the rate in patients whose albumin levels before therapy were between 3.8 g/dl and 5.3 g/dl was 80.0%. The efficacy rate in patients whose neutrophil counts before therapy were less than 100/microliter was 50.0%, whereas the rate in patients whose neutrophil counts after therapy were less than 100/microliter was 53.8%. The efficacy rate in patients whose neutrophil counts both before and after therapy were less than 100/microliter was 37.5%. Side effect of exanthema was observed in 1 patient. These results indicate that CZOP is an effective and safe antibiotic for the treatment of febrile neutropenia in patients with hematological malignancies. PMID- 14750325 TI - [A case of pulmonary metastases from laryngeal cancer showing a complete response to chemotherapy with docetaxel]. AB - Docetaxel (TXT) is a new cytotoxic agent against head and neck, (H and N) cancer since the emergence of cisplatin. We herein describe the efficacy of TXT in treating distant metastases from H and N cancer. The patient was a 61-year-old man who had undergone laryngectomy and irradiation for laryngeal cancer (glottic type: T3N0M0). More than 2 years later after the first course of therapy, pulmonary metastases involved in the left lower area were revealed by lung CT. The divided TXT therapy (20 mg, 3 times, and 40 mg, 3 times) was carried out without any sequelae. This consecutive therapy resulted in complete disappearance of the metastases on follow-up CT. It is suggested that divided TXT therapy has potential as a new treatment modality for distant metastases of H and N cancer. PMID- 14750326 TI - [Metastatic oropharyngeal cancer successfully treated with docetaxel, cisplatin, 5-FU and l-leucovorin]. AB - We encountered a patient with metastatic disease of the lung from oropharyngeal cancer who achieved a complete response to 3 cycles of chemotherapy with docetaxel, cisplatin, 5-FU and l-leucovorin (TPFL). A 72-year-old man was found to have metastatic disease of the lung 32 months after treatment of oropharyngeal squamous cell carcinoma. Chemotherapy was initiated with TPFL and the patient obtained a complete response after 3 cycles. Twelve months after chemotherapy he had no recurrence. We conclude that the use of docetaxel cisplatin, 5-FU and l leucovorin for metastatic squamous cell carcinoma of the head and neck is a viable option. PMID- 14750327 TI - [Complete remission of locally advanced squamous cell lung cancer induced by chemotherapy with cisplatin and docetaxel]. AB - The patient was a 70-year-old man with c-T2N3M0 stage IIIB squamous cell lung cancer in the upper lobe of his left lung. Two cycles of chemotherapy of docetaxel with cisplatin induced complete response. Sequential radiotherapy was performed for the mediastinum. One year after treatment, he underwent a lung resection for a second lung cancer, poorly differentiated adenocarcinoma, in the upper lobe of his right lung. Resected mediastinal lymph nodes showed no cancer cells of the first lung cancer. Now, more than 3 years after the first examination, he maintains complete remission without evidence of recurrence. PMID- 14750328 TI - [A case of peritonitis carcinomatosa of pulmonary adenocarcinoma in which gefitinib was effective]. AB - We report a case in which gefitinib was effective in reducing the amount of ascites. A 61-year-old man with peritonitis carcinomatosa due to metastasis from pulmonary adenocarcinoma was admitted to our hospital. Various chemotherapies were not effective for controlling his ascites. Therefore, we orally delivered gefitinib at a dose of 250 mg/day. Three days after administration, a reduction in the amount of ascites was noticed. Subjective symptoms including general fatigue and difficulty breathing were dramatically improved as was QOL. We conclude that this case was a good candidate for treatment using gefitinib. PMID- 14750329 TI - [A case of effective weekly paclitaxel administration for primary lesion of schirrous gastric cancer]. AB - We report a case of advanced schirrous gastric cancer with carcinomatous peritonitis. Chemotherapy with TS-1 was applied during the first 4 weeks, but the tumor did not respond to this therapy. Next, paclitaxel (TXL) was administered at a weekly dose of 90 mg/body/day for 3 weeks followed by a week interval of rest. Remarkable mass reduction of primary tumor was observed after 3 courses of treatment, and the symptom derived from primary tumor was relieved without significant side effects. The clinical course for schirrous gastric cancer with carcinomatous peritonitis is still miserable for patients, although many attempts have been made to improve its prognosis. A weekly paclitaxel regimen appears to be one of the promising treatment for schirrous gastric cancer. PMID- 14750330 TI - [A case of gastric cancer with multiple liver metastases and spleen metastasis that responded to low-dose CDDP/TS-1 combination therapy]. AB - Since the introduction of TS-1 for clinical treatment of the progression or recurrence of stomach cancer, the effectiveness of combination therapy incorporating other agents with CDDP has been reported. Low-dose CDDP/TS-1 combination treatment was carried out in a case of Stage IV progressive stomach cancer showing multiple liver metastases and spleen metastasis. Regression of the primary carcinoma and reduction in size of liver metastases and spleen metastasis were observed. Grade 2 leukocyte decrease and grade 1 stomatitis were noted as adverse reactions to the treatment. Low-dose CDDP/TS-1 combination therapy was useful in this case of advanced gastric cancer. PMID- 14750331 TI - [A case of malignant gastric outlet stenosis palliated with self-expandable metallic stent and responding to TS-1, an oral fluorouracil derivative]. AB - We report a case of advanced gastric cancer with antral stenosis that responded to an oral anticancer agent, TS-1, after metallic stent insertion. A 59 year-old man was admitted to our hospital because of abdominal distension and vomiting after meals. The diagnosis was advanced gastric cancer with antral stenosis and multiple liver metastases. FP therapy (CDDP 80 mg/m2 day 1 div, 5-FU 700 mg/m2 day 1-5 continuous div) was administered. Although endoscopic findings showed improvement after the first course, the condition of the patient did not improve. We therefore inserted a self-expandable metallic stent into the antral stenosis. After implantation, the patient was able to have regular meals, leave the hospital and return to work. TS-1 (120 mg daily), an oral fluorouracil derivative, was administered in the outpatient setting. A partial response (PR) was obtained after 2 courses with regression of multiple liver metastases and the primary tumor. PMID- 14750332 TI - [A case of recurrent advanced gastric cancer with lung metastasis effectively treated by combined chemotherapy of TS-1 and weekly CDDP]. AB - We report a case of a patient with recurrent gastric cancer and lung metastasis, who responded remarkably to combination chemotherapy using TS-1 and weekly CDDP. The patient was administered 2 courses of TS-1 (80 mg/m2/day, on day 1-21) and CDDP (25 mg/m2/day, on day 8, 15, 22) every 5 weeks. The regimen was done on an outpatient basis. The treatment resulted in the metastatic tumors in the lung disappearing after 1 course. No severe side effects were observed. This combination therapy proved useful for treating lung metastasis from gastric cancer in this patient. PMID- 14750333 TI - [Complete response to treatment with low-dose FP therapy in a patient with stage IVB primary hepatocellular carcinoma with multiple lung and bone metastases]. AB - We report a case in which low-dose FP (5-fluorouracil/cisplatin, 5-FU/CDDP) therapy was remarkably effective for stage IVB advanced hepatocellular carcinoma (HCC) with lung and bone metastases. 5-FU of 250 mg/body/day was continuously infused over 24 hours and CDDP of 10 mg/body/day was infused over 30 minutes from day 1 to day 5 in a week. Administration was continued for 4 weeks as 1 cycle. An 81-year-old woman was diagnosed with HCC in S3 and underwent a transcatheter hepatic arterial embolization (TAE) for the tumor in December 2000. The patient complained of lumbago and hip pain in July 2001 and was admitted for dysbasia in September 2001. On admission, the level of serum AFP and PIVKA-II elevated to a remarkable 59,300 ng/ml and 25,700 AU/ml. Chest computed tomography (CT) showed multiple bilateral lung metastases and abdominal CT showed a tumor 12 x 11 x 10 cm in diameter in the right, iliac bone. No recurrent sign was found in the liver except for the accumulation of Lipiodol. Low-dose FP therapy of 2 cycles was performed. The levels of serum AFP and PIVKA-II decreased to 374 ng/ml from 59,300 and to 35 AU/ml from 25,700, respectively, after this therapy. The CT findings revealed that a complete response (CR) was obtained for lung metastases and a partial response (PR) was obtained for bone metastases after completion of course 2, and maintained thereafter. The oral UFT of 600 mg was administered after completion of course 2 in the outpatient setting. The level of AFP and PIVKA-II decreased to 13.2 ng/ml and to 26 AU/ml, respectively, in February 2002. No sign of recurrence was seen during the 13 months of follow-up after low-dose FP therapy. Toxic events consisted of only leukopenia (grade 1). Her quality of life (QOL) was fair during this therapy. Low-dose FP therapy is possibly useful for patients with stage IVB advanced HCC. PMID- 14750334 TI - [A case of liver metastatic recurrence of bile duct cancer completely responding to single drug, UFT, chemotherapy]. AB - A 72-year-old man with common bile duct cancer was treated by pylorus preserving pancreatoduodenectomy with D3 lymph node dissection and preventive radiotherapy at hepaticojejunostomy. At 4 months after surgery, CT demonstrated multiple liver metastatic tumors. Single drug chemotherapy, UFT 600 mg, was started. After 6 months, the liver metastatic tumor could not be seen by CT. UFT may be a first candidate for chemotherapy for recurrence of bile duct cancer. PMID- 14750335 TI - [A case of rectal cancer with distant lymph node metastases completely responding to postoperative chemotherapy with levofolinate combined with 5-fluorouracil]. AB - The patient was a 58-year-old female who had a cancer of the rectum with multiple lymph node metastases including paraaortic lymph nodes and Virchow lymph nodes. Abdominoperineal resection was performed palliatively. After the operation, weekly bolus of 5-fluorouracil combined with levofolinate was carried out. After 2 courses of chemotherapy, metastases of paraaortic lymph nodes and Virchow lymph nodes completely disappeared upon CT examination. Chemotherapy has continued for 14 months, and she is well and has maintained a complete response for more than 1 year. This case suggests that this combination chemotherapy of LV/5-FU, admitted in Japan, is effective against advanced colorectal cancer. PMID- 14750336 TI - [Historical background and current status of the researches on sentinel node concept]. PMID- 14750338 TI - Facility profile. Design takes flight at Florida hospital. PMID- 14750337 TI - [Molecule based diagnosis]. AB - Based on reviews of the concept of diagnostics and in general and in specific tumour areas it was clear that development of diagnostic procedures involving genomics will allow for much better targeted and tailored treatments in the future. This will result in better efficacy and better tolerability of cancer treatments, but will also allow for progress in prediction, diagnosis and dose selection. Large collaborative projects studying the efficacy and safety of drugs on the genome level is promising to bring important benefits to both patients and the national economy by reducing useless drug therapy. In colorectal cancer there are several genetic defects identified that can act as the target for directed therapy in the future. Expressions of tumour specific antigens open the way for immunological targeted therapies. Developments in the understanding of the genomic basis for resistance to anti-tumour therapy is promising to help targeting patients likely to respond and not develop resistance. A Japanese model is being developed to determine the relative risk of breast cancer of Japanese women. Based on this prevention therapies can be instigated. The last four years have seen the introduction of four novel targeted therapies. If this model should become a standard in the future, much stronger collaboration between academic research and pharmaceutical industry need to develop. PMID- 14750339 TI - Planning for PACS. Imaging upgrades present infrastructure challenges. PMID- 14750340 TI - Growing issue. An expert's perspective on molds + legal discovery. PMID- 14750341 TI - Water watch. A look at water quality recommendations + requirements. PMID- 14750342 TI - Trends in critical care planning and design. Panel discussion. PMID- 14750343 TI - New year, 'new pathways'. Are you ready for the JCAHO's new survey process? PMID- 14750344 TI - Microfibers, macro benefits. Health care facilities discover microfiber mops and cloths. PMID- 14750345 TI - Facial pain and sinonasal surgery. AB - OBJECTIVES: To examine the causes of facial pain that persists after endoscopic and other sinonasal surgery. STUDY DESIGN & SETTING: A study of a cohort of 973 patients presenting in the outpatient clinic with symptoms of facial pain and/or rhinosinusitis. The study subgroup consisted of 75 patients with facial pain persisting after sinus surgery (endoscopic procedure n = 48, other intranasal procedures n = 27). The patients in the subgroup were studied with particular reference to their original presenting symptoms, endoscopic signs, and findings at CT as well as their symptoms when they presented to this unit and, importantly, these were analysed in the context of their response to treatment and follow-up after a mean of 2 years 7 months. RESULTS: Of the 75 patients who had persistent pain after surgery, 40 had no evidence of disease at endoscopy or CT at their initial presentation, whilst 35 had preoperative evidence of sinonasal disease. However, after surgery and re-evaluation, as well as neurological medical treatment, it was found that sinonasal disease was not the cause of these patients pain. The causes of their facial pain that persisted after sinonasal surgery were tension-type headache, atypical facial pain, migraine, paroxysmal hemicrania, cluster headache but the most common cause was 'midfacial segment pain', which has all the characteristics of tension-type headache but primarily affects the midface. CONCLUSIONS & SIGNIFICANCE: Some patients with facial pain are undergoing endoscopic sinus surgery in the mistaken belief that rhinosinusitis is the cause of their facial pain. All surgeons dealing with facial pain should be familiar with non-sinonasal diagnoses. A new category of facial pain, 'midfacial segment pain', is described and proposed. PMID- 14750346 TI - Nasal polyps and middle turbinates epithelial cells sensitivity to amphotericin B. AB - HYPOTHESIS: Intranasal application of the antimycotic agent amphotericin B (AmphoB) has been proposed as an effective treatment of chronic rhinosinusitis (CRS) with polyps. AmphoB is a sterol-binding agent known to modify cell membrane structure. The cytotoxic effects of AmphoB were studied on primary human nasal epithelial cells in vitro. METHODS: Human epithelial cells were isolated from nasal polyps and middle turbinates of patients suffering from CRS, and grown on collagen-coated polycarbonate filters with an air liquid-interface. After 15 days of culture, cells were exposed apically to 50 microM AmphoB during 4 h daily for 5 days. Some cells were treated during 4 weeks. The bioelectric properties of cells were then studied in Ussing chambers. Integrity of the cell monolayers was assessed by measurement of the transepithelial resistance (R) and immunofluorescent localization of the tight junction protein occludin. RESULTS: Disruption of the epithelial monolayer integrity was observed in all of the nasal polyps cell cultures, as demonstrated by a 60% drop in R. Immunofluorescence microscopy showed significant loss in cell number and disruption in the distribution of occludin. Turbinate cell cultures elicited no change in R and expression of occludin after AmphoB treatment. However, the transepithelial potential, the basal short-circuit current and the amiloride-sensitive current were reduced by 70%. CONCLUSIONS: AmphoB was cytotoxic for nasal polyp epithelial cells with disruption of the epithelium integrity and loss of tight junctions. In contrast, integrity of turbinate epithelial cells was conserved despite alterations in transepithelial ion transport. These observations may explain the beneficial effect of intranasal application of AmphoB on CRS observed in clinical trials. PMID- 14750347 TI - Peak nasal inspiratory and expiratory flow measurements--practical tools in primary care? AB - General practitioners diagnose and treat numerous patients with nasal problems but have no objective equipment to aid diagnosis. We studied the characteristics of peak nasal inspiratory (PNIF) and expiratory (PNEF) flows to explore their utility for primary care use. One hundred healthy volunteers, 50 men, 50 women, performed PNIF and PNEF measurements in standardised laboratory circumstances. Repeatability was tested for a subgroup of 20 women who performed two consecutive series of PNIF and PNEF at a 2-minute interval. Diurnal variability was tested with a subgroup of 10 men and 10 women who recorded PNIF and PNEF values at home every morning and evening for 7 days. Distribution of individual values for both PNIF and PNEF was wide and independent of subject's age, height, or gender. Repeatability was poor and diurnal variation substantial. PNIF and PNEF lack the properties of good clinical tests and thus cannot be recommended for primary care use. PMID- 14750348 TI - Evolution of symptoms associated to nasal polyposis following oral steroid treatment and nasalization of the ethmoid--radical ethmoidectomy is functional surgery for NPS. AB - PURPOSE: The effects on the symptoms of nasal polyposis (NPS) of 1) a 7-day systemic steroid treatment and 2) radical ethmoidectomy (nasalisation) were compared. PATIENTS AND METHODS: Twenty-four patients with NPS whose symptoms failed to respond to medical treatment were included in the study. Symptoms were recorded on Visual Analog Scales (VAS) before (Q1) and the day (Q2) after a 7-day treatment of oral prednisolone (60 mg/day), and a few weeks later the day before surgery (Q3). All patients were operated on bilaterally according to the nasalization principles, i.e. endoscopic radical ethmoidectomy without mucosal preservation, with middle turbinate resection, antrostomy, sphenoidotomy and frontal ostium exposure, and a depot injection of triamcinolone 80 mg the day after surgery. The fourth questionnaire (Q4) was fulfilled one month after surgery, the day of the first postop visit. The following questionnaires were returned by mail at 3 months (Q5), 6 months (Q6), 9 months (Q7), and 12 months (Q8) post-operatively. Patients stayed on topical steroids throughout the study. RESULTS: Nasal obstruction was a major complaint at entry in the study. Following the short-systemic steroid course the obstruction score improved significantly. However, at 2 months after the oral steroid treatment the obstruction score had deteriorated again. Following surgery, obstruction scores ameliorated again and remained stable over the full year of follow-up. Similar results were observed for anterior and posterior rhinorrhea, sneezing and itching. None of the patients reported any intake of systemic steroids during follow-up. CONCLUSION: These data show that 'nasalization' i.e. radical ethmoidectomy with middle turbinate resection and mucosa removal is effective functional surgery for patients with nasal polyposis if medical treatment fails. The subjective effects on the sense of smell are reported in a separate paper published in this issue. PMID- 14750349 TI - Olfaction in patients with nasal polyposis: effects of systemic steroids and radical ethmoidectomy with middle turbinate resection (nasalization). AB - AIM: In this prospective study the effect of medical and surgical treatment on subjective olfaction was studied in patients with nasal polyposis (NPS). The effects on nasal obstruction, anterior and posterior rhinorrhea, sneezing and itching are reported in another article in this issue. PATIENTS AND METHODS: Protocol 1. Twenty-four patients with NPS who complained about anosmia were treated with a 7-days course of systemic steroids. Their subjective overall sense of smell was determined with a visual analog scale (VAS) before treatment, immediately after treatment, and two months later. Subsequently all patients underwent surgery bilaterally according to the nasalization principles. The sense of smell was re-evaluated at 1, 3, 6, 9, and 12 months postoperatively. Protocol 2. Thirty-two patients with NPS not responding to medical therapy who, for different reasons, did not receive oral steroid treatment, received surgery only and were followed up during one year after nasalization. Of these patients, 25 were anosmic and 7 normosmic. RESULTS: Protocol 1. Following the 7-day treatment with systemic steroids the olfactory score increased significantly. During the waiting time for surgery (64 +/- 39 days) this score deteriorated again in a significant way. One month after nasalization which included a depot injection of triamcinolone 80 mg the day after surgery, the olfactory score ameliorated again and remained stable at 3, 6, 9, and 12 months. None of the patients reported any intake of systemic steroids during the one-year of follow-up. Statistically, there was a trend suggesting that the 12 month post-nasalization score was better than the immediate post-oral steroid score. A good correlation between the improvement of the sense of smell after 7 days of systemic steroids and one year after nasalization was found. Protocol 2 One month after the nasalization protocol, olfaction in patients of the hypo-anosmic group had improved considerably. Scores at 3, 6, 9, and 12 remained very stable. The sense of smell in the normosmic group did not change after surgery and remained stable during the year of follow-up. In total, 49 patients with a severe loss of smell showed a significant improvement at 12 months after surgery. CONCLUSION: The present study shows that 1) long-lasting correction of olfactory dysfunction produced by nasal polyposis can be achieved through the combination of nasalization and low dose of nasal steroids, 2) middle turbinate resection does not alter the possibilities to restore the sense of smell. PMID- 14750350 TI - Three-wall decompression technique using transpalpebral and endonasal approach in patients with Graves' ophthalmopathy. AB - PURPOSE: To evaluate the effects of the three-wall decompression technique using transpalpebral and endonasal approach in patients suffering from Graves' ophthalmopathy. METHODS: In this prospective study, we present a consecutive series of 15 subjects (17 eyes) who were submitted to orbital decompression by removing the inferior and lateral walls using transpalpebral incision combined with a transnasal endoscopic resection of the medial wall. The surgical technique involved the preservation of the bone structure between the lamina papyracea of the ethmoid and the maxillary orbital floor. MAIN RESULTS: The mean ocular recession based on Hertel measurements was 6.00 mm (range, 4-9 mm). None of the patients presented pre-operation diplopia, and one developed post-operation diplopia. Visual acuity was preserved in all cases. CONCLUSION: It is safe and efficient to perform three-wall decompression, combining transpalpebral and endoscopic transnasal approach, with preservation of the bone structure and the bone lateral to the infraorbital canal with fixation by two titanium plates on the lateral edge and removal of intraorbital fat, which results in significant proptosis reduction and minimal complications. PMID- 14750351 TI - Juvenile nasopharyngeal angiofibroma: the routes of invasion. AB - The juvenile nasopharyngeal angiofibroma has a characteristic growth in all directions from its origin. However, the extensions of the tumor seem to be independent, each one with distinct behavior. The aim of this study is to analyze the preferential direction and routes of JNA growth, as well as its correlation with the patient's age. We analyzed 33 patients without any previous treatment, attempting to the extension and routes of tumor's growth (CT scan), and its correlation with the patient's age. The sphenopalatine foramen region was affected in all cases. From this point, a growth towards several routes with a different rhythm was noted, determining variable configurations to the tumor. The lateral and superior growths were the most frequent. The expansion into the pterygopalatine fossa was very frequent and could involve important anatomical structures, determining higher morbidity. Three sites were invaded through more than one route: pterygoid fossa, middle cranial fossa and maxillary sinus. There was no significant correlation between invasion route and patient's age. However, considering the age, there was a concomitance between tumor development and facial growth by "displacement". We discuss this condition, suggesting an explanation to the tumor invasion and expansion inside the pterygopalatine fossa. PMID- 14750352 TI - Double-blind randomised controlled trial comparing Merocel with Rapid Rhino nasal packs after routine nasal surgery. AB - INTRODUCTION: Nasal packing after routine nasal surgery is commonly practiced in the UK. The most popular pack appears to be Merocel, however this can be associated with significant pain on its removal. AIM: To test the efficacy of a relatively new nasal pack--the Rapid Rhino Goodman pack--introduced to our department, which claims to reduce pain and bleeding after nasal surgery. METHODS: Fourteen patients undergoing routine nasal surgery were recruited into a randomised controlled trial. One nasal cavity was packed with a Merocel pack and the other with a Rapid Rhino Goodman pack. Patients were asked to record pain levels on each side using a visual analogue scale. RESULTS: We found no difference between packs whilst in-situ, with all patients recording low to moderate pain scores. There were significantly higher pain levels associated with Merocel pack removal than with Rapid Rhino pack removal (average pain scores 5.64 vs 1.64, p < 0.001) and less bleeding overall. CONCLUSIONS: Rapid Rhino (Goodman pack) is associated with significantly less pain on removal than Merocel pack, causes less bleeding and is not more painful whilst in-situ. We therefore recommend its use in routine nasal surgery. PMID- 14750353 TI - Hemostasis in endoscopic sinus surgery using a specific gelatin-thrombin based agent (FloSeal). AB - The safety and efficacy of a new hemostatic sealant, based on a gel with collagen derived particles and topical thrombin (FloSeal, Fusion Medical Technologies, Inc. Fremont, CA) were assessed as an alternative to nasal packing for hemostasis in functional endoscopic sinus surgery. In a prospective clinical study of 50 patients undergoing bilateral endoscopic anterior ethmoidectomy, 2 ml FloSeal was used after surgery to stop bleeding. The results were compared to a control group of 50 patients with Merocel packing and showed that intraoperative hemostasis was rapid and equal in both groups. The main advantages of the new hemostatic sealant included a higher degree of comfort during postoperative nasal breathing and absence of complaints due to pressure or pain. There was only one case of postoperative bleeding on the 6th day, which required nasal packing. There were no more cases of stenoses or synechia in the ostiomeatal complex than were found in the Merocel group. No systemic side effects due to FloSeal were observed. This specific hemostatic sealant was shown to be a safe and efficacious alternative method for hemostasis in endoscopic sinus surgery with high patient satisfaction and an easy and fast mode of application. PMID- 14750354 TI - Psammomatoid ossifying fibroma. AB - Psammomatoid ossifying fibroma (POF) represents a unique subtype of fibro-osseous lesions. We describe a case of POF involving the orbit and the sinonasal tract, in a 13-year-old white female. Diagnosis depends on the histological, radiological and clinical features. Complete excision by endoscopic nasal surgery was the treatment of choice. Five years later the patient was free from symptoms and tumour recurrence. Differential diagnoses are discussed. PMID- 14750355 TI - Adenoid cystic carcinoma of the nasal septum. AB - Adenoid cystic carcinoma is a malignant tumour frequently described arising from seromucinous salivary tissue in the major and minor salivary glands. Within the nasal cavity, it is uncommon and usually involves the lateral wall. A rare case of adenoid cystic carcinoma of the nasal septum is presented along with a review of the literature. The presentation and management of this uncommon condition is discussed. PMID- 14750356 TI - A morphometric consideration of nasal septal deviations by people with paranasal complaints; a computed tomography study. PMID- 14750357 TI - [Brief intervention for smoking, problem drinking and drug abuse by high school students]. AB - Adolescent smoking, drinking and illegal drug abuse (drug-related problems) in Japan were treated as a legal model in the second prevention area. This study showed the necessity and usefulness of early intervention concerning adolescent drug-related problems as adolescent mental health problems. First, school nurses were asked if they were consulted about drug-related problems by students. Many school nurses in junior and senior high schools were consulted about drug-related problems by students, and they emphasized the need for counseling systems for drug-related problems by professionals. The second part of the study involved brief interventions by the authors in high school students with a high risk of drug-related problems. The authors went to three high schools and conducted brief interventions on students who had drug-related problems and sought advice. We prepared several screening tests for assessment of drug-related problems, such as the Fagerstrom Test for Nicotine Dependence (FTND). Adolescent Alcohol Involvement Scale (AAIS). Quantity-Frequency Scale (QF Scale). Core Alcohol Use Disorders Identification Test (Core AUDIT), CAGE and Drug Abuse Screening Test-20 (DAST-20). In the brief interventions, the authors assessed drug-related problems of students using these scales and advised them how to revolve such problems two or three times. Twenty-two students accepted brief interventions. All of them smoked, 62% of them showed problem drinking and 33% illegal drug use. At 6 months after the brief interventions, 16 out of 21 students reported their drug-related problems. Nineteen percent of them had reduced smoking, 67% of them decreased drinking and one half of them decreased their use of illegal drugs. Results of this study revealed the necessity and the usefulness of the brief intervention for high school students with drug-related problems. PMID- 14750358 TI - Adolescent smoking behavior in Japan, 1996. AB - We conducted second nationwide survey on smoking behavior among Japanese high school students. The survey was a cross-sectional sampling survey. The survey's targets were junior and senior high schools throughout Japan. Sample schools were selected by stratified cluster sampling. Self-administered anonymous questionnaires were sent to sample schools for all students to fill out. 65.6% of the junior high schools and 67.0% of the senior high schools were responded to this survey. A total of 117,325 students responded and 115,814 questionnaires were subjected to analysis. 7.5% of boys and 3.8% of girls in the 1st grade of junior high school were classified as current smokers, compared with 36.9% of boys and 15.6% of girls in 3rd grade of senior high school. Less than 1% of boys and girls in the 1st grade of junior high school were classified as daily smokers, this figure increased markedly by the 3rd grade of senior high school to reach 25.4% of boys and 7.1% of girls. Of students in the 3rd grade of senior high school, 21.2% of boys and 9.9% of girls had tried smoking before 13 year of age. Cigarette consumption also increased with age, although the proportion of those smoking 20 or more cigarettes per day did not. The major source for cigarettes among current smokers was vending machines. This survey revealed that smoking prevalence among Japanese high school students has already reached quite high level. Comprehensive energetic measures for control minor smoking are necessary in Japan. PMID- 14750359 TI - [A case of female alcoholic who developed liver cirrhosis with small amounts of alcohol by the common use of contraceptive agent]. AB - A case of female alcoholic who developed liver cirrhosis with small amounts of alcohol by the common use of contraceptive agent was reported. A case was a 33 year-old female who had complained of systemic edema and jaundice. She had been drinking alcohol, while she had been taking the contraceptive agent from 20-year old. On admission, she had a large amount of ascites with jaundice. She was diagnosed as liver cirrhosis and hepatic failure by CT scanning of abdomen and laboratory data. Her condition was temporarily improved by the abstinence and the treatment. Since she drank under hospitalization, she had to change the hospital and died after 2 months. She had been drinking for only 10 years. Her cumulative alcohol intake was also very small. She may have developed alcoholic cirrhosis with small amount of alcohol because of common use of contraceptive agent with drinking. PMID- 14750360 TI - Brain excitatory amino acid transporters (EAATs) and treatment of methamphetamine toxicity. AB - Based on the phenomenon of the abnormally increased transport of brain excitatory amino acids induced by the increased release of dopamine (DA) in the brain, the effects of intraperitoneal L-trans-pyrrolidine-2,4-dicarboxylic acid (PDC), a non selective excitatory amino acid transporter (EAAT) inhibitor, and (+/-)-threo-3 methylglutamic acid (MG), a specific EAAT2 inhibitor, were examined against methamphetamine (MA) and cocaine (COC) toxicity in mice. The MA (5 mg/kg) increased activity counts, which included counts of both ambulatory and stereotyped behaviors, were attenuated by 10 and 20 mg/kg of PDC, but the COC (40 mg/kg)-increased activity counts were attenuated only by 20 mg/kg PDC. PDC (20 mg/kg) significantly attenuated both the mortality rate and the seizure score in acute MA (18 mg/kg) toxicity, but attenuated only the seizure score in acute COC (75 mg/kg) toxicity. PDC and MG (repeated doses of 5 and 10 mg/kg) attenuated the mortality rate (significant attenuation in the PDC group) and seizure score against repeated MA (12 mg/kg) toxicity, but had no effect on repeated COC (60 mg/kg) toxicity. Furthermore, MA (5 mg/kg) and COC (40 mg/kg) induced stressor like and anxiogenic effects, the former of which were attenuated by PDC only (10 and 20 mg/kg in the MA group and 20 mg/kg in the COC group), and the latter of which were attenuated by both PDC and MG (for both drugs, 10 and 20 mg/kg in the MA group and 20 mg/kg in the COC group). Therefore, it was concluded that not only EAAT2 but also the other EAATs contributed to the occurrence of the MA induced effects and part of the COC-induced effects, and that a non-selective EAAT inhibitor notably blocked the behavioral effects accompanying the MA-induced over-release of DA. PMID- 14750361 TI - [Difference between alcohol dependence and nicotine dependence in cognitive dysfunction]. AB - The extent of cognitive dysfunction in the nicotine and/or alcohol dependent subjects has been evaluated with Digit Span, Number Connection Test and Wisconsin Card-Sorting Test. Samples were divided into 4 groups, 8 subjects with alcohol and nicotine dependence, 9 subjects with alcohol dependence, 10 subjects with nicotine dependence and 31 subjects without the dependences who met ICD-10 criteria for dependence syndrome. The performance times of Number Connection Test were prolonged in the alcohol dependence and the presence of the nicotine dependence did not influence the performance times of Number Connection Test. The performances on the Wisconsin Card-Sorting Test were impaired by the alcohol dependence and the presence of the nicotine dependence did not influence the performances on the Wisconsin Card-Sorting Test. PMID- 14750362 TI - [Requisition of the revised criteria on infectious wastes disposed from medical institutions]. PMID- 14750363 TI - [Effects of dietary variety on declines in high-level functional capacity in elderly people living in a community]. AB - OBJECTIVE: The purpose of the investigation was to assess effects of dietary variety on declines in high-level functional capacity in community dwelling elderly, based on a 5-year longitudinal study (from 1992 to 1997). SUBJECTS AND METHOD: Subjects were a representative sample comprising 235 men (mean age 70.8 years) and 373 women (71.7 years) aged 65 years and above, living in Nangai village in Akita prefecture. Baseline and follow-up surveys were undertaken by the interview method. For assessing dietary variety, we introduced the dietary variety score (DVS), counting the number of 10 food-groups consumed daily from food frequency questionnaires: meat, fish and shellfish, eggs, milk, soybean products, potatoes, green yellow vegetables, fruits, seaweed, and fat and oil. The DVS ranged from 0 to 10 with higher score indicating a higher dietary variety. We also evaluated higher-level functional capacity using the Tokyo Metropolitan Institute of Gerontology Index of Competence (TMIG Index of Competence). Effects of dietary variety on declines in TMIG Index of Competence were analyzed by multiple logistic regression analysis. RESULTS: The average TMIG Index of Competence score for men and women were 12.1 and 10.8 at baseline, respectively. The mean baseline DVS was 6.3 for men, and 6.2 for women. Relative to the reference groups with DVS in 1-3, the groups with 4-8 and 9-10 scores had lower risks for decrease in TMIG Index of Competence scores over the study period. Relative risks (95% confidence interval) of the groups with a DVS of 4-8 and 9-10 were 0.92 (0.50-1.67) and 0.71 (0.34-1.48) regarding instrumental self maintenance, 0.50 (0.29-0.86) and 0.40 (0.20-0.77) for intellectual activities, and 0.44 (0.26-0.75) and 0.43 (0.20-0.82) for social roles of sub-scales of TMIG Index of Competence, adjusting for age, sex, educational attainment, and the baseline TMIG Index of Competence scores. CONCLUSION: Higher dietary variety is associated with a reduced risk of higher-level functional decline in community dwelling elderly. PMID- 14750364 TI - [Relationship of childhood obesity to adult obesity: a 20-year longitudinal study from birth in Ishikawa Prefecture, Japan]. AB - OBJECTIVE: Evidence regarding the relationship between childhood obesity and adult obesity in Japan is limited. This study was conducted to determine the relationship between childhood mass index (BMI) at 3 months, 12 months, 3 years and 20 years in a general population. METHODS: Data obtained from men and women aged 20 years (born between 1968-1974), who had received medical examinations in Ishikawa Prefecture, Japan, were linked to data of medical examinations of the same individuals as infants (3 months, 12 months, and 3 years). The relationship between childhood BMI (or Kaup index) and adult BMI was analyzed for a total of 2,314 participants (1,080 men and 1,234 women), whose data could be followed for 20 years. RESULTS: BMI at 20 years displayed significant positive correlations with BMI at 3 months, 12 months, and 3 years; this correlation was strongest with respect to BMI at 3 years (r = 0.33, p < 0.001 in men; r = 0.42, p < 0.001 in women). In terms of percentages of obese participants (BMI 25 kg/m2 or over) at 20 years in accordance with BMI categories at each age, the rates were 4.6% in men and 1.0% in women with a BMI less than 15 kg/m2 at 3 years, but 29.1% and 29.5%, respectively, with a BMI of 18 kg/m2 or over (6.3 and 29.5 times higher, respectively). Percentages of obese participants at 20 years were highest in those exhibiting an above average BMI at 3 years, regardless of the BMI at 3 months. CONCLUSIONS: Body mass in young adults is strongly related to body mass in childhood, especially with that at 3 years. About 30 percent of obese children at 3 years remain obese into adulthood. These results are of interest with respect to assessment of future risk of adulthood obesity at medical examinations for infants in Japan. PMID- 14750365 TI - [Estimation of disease-specific costs in a dataset of health insurance claims and its validation using simulation data]. AB - PURPOSES: To estimate disease-specific costs in a dataset of health insurance claims with multiple diagnoses with known aggregate cost per claim and unknown disease-specific cost of each diagnosis using PDM (Proportional Disease Magnitude) method, validate its accuracy using simulation data with Monte Carlo method and improve its accuracy by developing an adjustment formula. METHODS: Developed simulation data with pre-assigned disease-specific costs, applied PDM method using arithmetic means of per-diem-per-disease cost as magnitude, validated its accuracy by observing the correlation between estimates by PDM method and known disease-specific costs and formulated an adjustment formula to improve accuracy. The reproducibility of the findings was assessed using Monte Carlo method by repeating the same procedures. RESULTS: The observed arithmetic means of per-diem-per-disease cost did not match well with actual values resulting in unsatisfactory accuracy. However, when the observed means were adjusted with a formula in which the observed mean is multiplied by (observed mean/overall mean) in the power of 2, PDM method yielded an accurate estimate of disease-specific cost. The accuracy was reproduced by Monte Carlo method with 0.9 or above R square value and slope of regression line in 76, 56 out of 100 iterations respectively. CONCLUSIONS: PDM method proved to be an objective, reproducible and accurate method for estimation of disease-specific costs of health insurance claims. PMID- 14750366 TI - [The relationship between dental caries of deciduous teeth and anxiety of mothers associated with child-care]. AB - AIM: The incidence of caries in children is closely related to daily habits and may also be influenced markedly by a mother's attitude to child-care. For example, if a mother has a mental health problem such as child-care anxiety, this may prevent her obtaining advice on the daily health requirements of her children including the need to brush the teeth regularly. It is therefore important that mothers receive guidance in order to minimize the incidence of caries. This study examined the effect of child-care anxiety in mothers on the prevalence of caries in 3-year-old children. METHODS: The subjects were 503 pairs of mothers and their children who attended a check-up at a community health center in Gifu City when the children were aged 3-years. The mothers answered a questionnaire about child care, while the children had a dental examination and caries activity test (Cariostat). The prevalence of caries and the Cariostat score were examined by binomial logistic regression analysis. The relationship between child-care anxiety of the mothers and children's caries or Cariostat score were analyzed using Covariance Analysis, that incorporated additional multiple indicator models including snack behavior, social-support and food faddism of the children. RESULTS: 1. The rate of caries disease increased with high Cariostat scores and was statistically significant at scores greater than 2.0. 2. Covariance analysis demonstrated both the models of children's caries and Cariostat score provided a good fit to the actual data, the former model having a GFI = 0.951 and RMSEA = 0.046, and the latter a GFI = 0.952 and RMSEA = 0.046. 3. It was found that anxiety over child-care directly affected snack behavior, while itself being influenced directly by both social-support and food fads of the children. 4. Snack behavior influenced Cariostat score, but not the prevalence of child caries. CONCLUSIONS: We suggest it is necessary to provide guidance to mothers with a background of child-care anxiety in order to prevent a high rate of caries in their children. It may be possible to detect mothers with child-care anxiety that leads to bad snack behavior by obtaining information on the child's social support and food fads. Our study showed that offering social support was an effective means of preventing habits that may predispose to the development of dental caries. Furthermore, we found the Cariostat score was an effective test for assessing the influence of child-care on caries incidence. PMID- 14750367 TI - [A survey on the status of medical treatment with high risk for infection among the users of home nursing stations--a comparison among services with and without medical facilities]. PMID- 14750368 TI - Traumatic deaths. PMID- 14750369 TI - Micronutrients and its correlation with mental performance among schoolchildren in Bario, Sarawak: a preliminary study. AB - Children who are born in a community with insufficient micronutrients, particularly iodine in remote rural areas are associated with low intellectual functions and mental retardation. The aim of this cross-sectional study is to determine the mental performance of the schoolchildren in Bario, a highland settlement of the Kelabit people in the district of Baram, Sarawak and to determine its correlation with the availability of iodine in the environment, as well as other micronutrients such as selenium, copper and manganese. A total of 25 schoolchildren in Bario age ranging from 7 to 12 years old participated in the study. Mental performance of the schoolchildren were tested using TONI-2 (Test of Nonverbal Intelligence--second edition), a cognitive ability measures with a response format which eliminates language and reduces motoric and cultural factors. The iodine levels from several wells, soils and salt found in Bario were determined using HPLC (software version 3.05.01) whilst serum levels of selenium, copper and manganese were measured using Graphite Furnace Atomic Absorption Spectrophotometer (GFAAS). The results showed that the median and mode scores of intelligence quotient for Bario were 82 and 75, respectively, whilst median and mode scores of intelligence percentile were 11.0 and 5.0, respectively. The maximum score achieved were at the average level with the quotient score between 90-110. It was found that salt produced from one of the wells in Bario contained high quantity of iodine. Based on standards established by the Trace Elements Laboratory, Roben Institute, University of Surrey, United Kingdom, schoolchildren in Bario are having sufficient blood levels of copper and a high selenium and manganese levels. Despite the remoteness of the study area, the schoolchildren in Bario, Sarawak showed higher mental performance compared to other isolated areas. This is probably correlated with the high micronutrients availability, particularly iodine, found naturally in Bario. PMID- 14750370 TI - Knowledge and practice of breast self examination and Pap smear screening among a group of electronics women workers. AB - A total of 486 Malaysian women electronics workers participated in a study of reproductive health knowledge and cancer screening. The practice of Breast Self Examination (BSE) was found to be related to educational attainment; while ever having had a Pap smear was found to be related to being older than 30 years old, being ever married, living with family or relatives, and not staying in hostels. Knowledge on reproductive health was found to be higher for older women, married women, living with family or relatives, not staying in hostels, ever having done BSE and ever having had a Pap smear. PMID- 14750371 TI - Stroke audit. AB - Medical audit is vital to ensure continuous quality assurance and quality improvements. We did a retrospective study to ascertain the adequacy of clinical documentation and the factors hindering early discharge after an acute stroke in a restructured hospital. The medical records of all patients with acute stroke who died or were discharged from a restructured hospital in Singapore in January and February 1999 were reviewed retrospectively. Demographic data and the presence or absence of clinical documentation were noted. Factors hindering the discharge of patients at Day 5, Day 10 of stroke and at final discharge were noted and classified into: stroke-related, complications of stroke, medical related and social factors. There were 101 patients in the study cohort, 55 males (54.5%) and 46 females (45.5%). The mean age was 67.9 years (SD 12.3). Documentation in Barthel scores (0%), presence of depression (0%), mental scores (1.0%), visual problems (10.0%), bladder continence (39.6%), admission functional status (37.6%) and dysphagia (52.5%) were deficient. The mean length of stay (LOS) was 13.0 (SD 14.2) days. The main factor hindering discharge at Day 5 (90.4%), Day 10 (95.2%) and at final discharge (82.1%) was stroke-related problems. Poor function (60.3%) and dysphagia (15.8%) were the 2 most common stroke-related problems hindering final discharge. Complications of stroke, medical-related problems and social reasons hindered final discharge in 10.8%, 17.8% and 2.9% of patients respectively. This audit revealed inadequacy in clinical documentation in patients with acute stroke. The main hindrance to final discharge of patients was stroke-related problems. The 2 most important stroke related problems were poor function and dysphagia. PMID- 14750372 TI - Gastrointestinal Cytomegalovirus infection in non-human immunodeficiency virus infected patients. AB - This is a retrospective study of fourteen patients who had proven Cytomegalovirus (CMV) infection of the gastrointestinal tract with no Human Immunodeficiency virus infection. The median age was 60.5 (Range 28 to 81) years. Eight patients were below (Group 1) and six above sixty five years old (Group 2). Areas of gastro-intestinal involvement were: oesophagus (2), stomach (1), colon (10) and multiple sites (1). Seven patients from Group 1 had received immunosuppressive therapy at the time of presentation and one had diabetes mellitus. We found a high prevalence of co-morbidities such as chronic renal failure and diabetes mellitus in Group 2. At median follow up of 13.9 months, there was a mortality rate of 50%. Only four patients were treated with ganciclovir. Our study concludes that the gastrointestinal CMV diseases in young patients were associated with immunosuppression whereas the older patients had chronic renal failure or diabetes. PMID- 14750373 TI - Protean manifestations of gastrointestinal tuberculosis. AB - We present 11 cases to illustrate the protean presentations of gastrointestinal tuberculosis. The patients presented with perianal fistula, appendicitis, ascites, rectal, intestinal or gastric 'growth', "ulcerative colitis", or recurrent anemia. In some of these cases there was no conclusive proof of infection with Mycobacterium tuberculosis but they responded well to empirical treatment with anti-tuberculous therapy. These cases illustrate that because signs and symptoms of intestinal tuberculosis are non-specific and even histology can be misleading, the diagnosis of gastrointestinal tuberculosis requires a high index of suspicion. A therapeutic trial of antituberculous drugs should be considered for patients with a high clinical suspicion of tuberculosis. PMID- 14750374 TI - Local experience in paediatric flexible bronchoscopy. AB - All children who underwent flexible bronchoscopy in the respiratory unit at Paediatric Institute, Hospital Kuala Lumpur from June 1997 to June 2002 were reviewed. A hundred and ten children underwent the procedure under sedation or general anaesthesia. The median age of these children was eight months. (Q1 3, Q3 30) The commonest indication for performing flexible bronchoscopy was for chronic stridor (50 cases) followed by persistent or recurrent changes such as lung infiltrates, atelectasis and consolidation on the chest radiographs (22). Laryngomalacia was found to be the commonest cause of stridor in 29 children. Two patients were diagnosed with pulmonary tuberculosis. With regard to safety, three procedures were abandoned due to recurrent desaturation below 85%. One of these patients had severe laryngospasm that required ventilation for 48 hours but recovered fully. Two neonates developed pneumonia requiring antibiotics following bronchoscopy. No patients developed pneumothorax or bleeding following the procedure. Bronchoscopy is a safe procedure when performed by well-trained personnel. Since it is an invasive procedure the benefits must outweigh the risks before it is performed. PMID- 14750375 TI - The sensitivity of the Malay version of Brief Manual of Sexual Function Inventory in assessing erectile dysfunction secondary to benign prostatic hyperplasia. AB - This study aimed to assess the sensitivity of the Malay version of the Brief Manual Sexual Function Inventory (BMSFI) on patients with and without urinary symptoms in Malaysian population. Validity and reliability were studied in patients with lower urinary tract symptoms (LUTS) and patients without LUTS. Reliability and validity was evaluated by using the test-retest method while internal consistency was assessed by Cronbach's alpha. Sensitivity to change was expressed as the effect size in the pre-intervention versus post-intervention score in patients who underwent transurethral resection of the prostate (TURP). Internal consistency was excellent. A high degree of internal consistency was observed for each of the 11 items and 5 domains (Cronbach's alpha value = 0.67 and higher and 0.73 and higher respectively). Test-retest correlation coefficient for the 11 items scores was highly significant. Intraclass correlation coefficient was high (ICC = 0.68 and above). The sensitivity and specificity showed a high degree of sensitivity and specificity to the effects of treatment. A high degree of significant level between baseline and post-treatment scores were observed across 3 domains in the treatment corresponds cohort but not in the control group. The Mal-BMSFI is a suitable, reliable, valid and sensitive to clinical change in the Malaysian population. PMID- 14750376 TI - Morbidity and process of care in urban Malaysian general practice: the impact of payment system. AB - BACKGROUND: The majority of primary care consultations in Malaysia occur in the general practice clinics. To date, there is no comprehensive documentation of the morbidity and practice activities in this setting. OBJECTIVES: We reported the reasons for encounter, diagnoses and process of care in urban general practice and the influence of payment system on the morbidity and practice activities. METHODS: 115 clinics in Kuala Lumpur, Ipoh and Penang participated in this study. General practitioners in these clinics completed a 2-page questionnaire for each of the 30 consecutive patients. The questionnaire requested for the following information: demographic data, reasons for encounter, important physical findings, diagnoses, investigations ordered, outpatient procedures performed, medical certificate given, medication prescribed and referral made. The morbidity (reasons for encounter and diagnoses) was coded using ICPC-2 and the medication data was coded using MIMS Classification Index. RESULTS: During 3481 encounters, 5300 RFEs (152 RFEs per 100 encounters) and 3342 diagnoses (96 diagnoses per 100 encounters) were recorded. The majority of the RFEs and diagnoses are in the following ICPC Chapters: Respiratory, General and unspecified, Digestive, Neurological, Musculoskeletal and Skin. The frequencies of selected aspects of the process of care (rate per 100 encounters) were: laboratory investigations 14.7, outpatient procedures 2.4, sick certification 26.9, referral 2.4, and medication prescription 244. Consultation for chronic diseases and acute infections were influenced more by demographic variables (age, employment) rather than payment system. Cash-paying patients were more likely to receive laboratory investigations and injections. CONCLUSION: This study demonstrated the breadth of clinical care in the general practice. Relatively fewer patients consulted specifically for preventive care and treatment of chronic diseases. The frequencies of outpatient procedures and referrals appeared to be low. Payment system results in important differences in patient mix and influences some types of practice activities. PMID- 14750377 TI - Trends in inquiries on poisoning: a five-year report from the National Poison Centre, Malaysia. AB - This report describes inquiries relating to poisoning cases which were received by the National Poison Centre of Malaysia from the years 1996 to 2000. The study utilized data from the NPC report forms. Only data relating to patient contact with a poison or chemical were included in the analysis. The poison centre received an average of 186 poisoning inquiries per year. Doctors remained the highest group of caller to the poison centre throughout the five-year period. Nearly 50% of all inquiries was regarding pesticide poisoning and this trend remained constant during the five-year period. Overall, the findings showed that poisoning inquiries relating to patient care were lower than in Japan and the United States. PMID- 14750378 TI - Cost analysis of cataract surgery with intraocular lens implantation: a single blind randomised clinical trial comparing extracapsular cataract extraction and phacoemulsification. AB - A randomised single blinded clinical trial to compare the cost of cataract surgery between extracapsular cataract extraction (ECCE) and phacoemulsification (PEA) was conducted at Hospital Universiti Kebangsaan Malaysia (HUKM) between March and December 2000. A total of 60 patients were included in this study. The cost of a cataract surgery incurred by hospital, patients and households up to two months after discharge were included. The costs of training, loss of patients' income after discharge and intangible costs were excluded. Results showed that the average cost for one ECCE operation is RM1,664.46 (RM1,233.04 RM2,377.64) and for PEA is RM1,978.00 (RM1,557.87-RM3,334.50). During this short period of follow up, it can be concluded that ECCE is significantly cheaper than PEA by an average difference of RM 313.54 per patient (p < 0.001). Cost of equipment and low frequency of PEA technique done in HUKM were the two main reasons for the high unit cost of PEA as compared to ECCE. PMID- 14750379 TI - Relationship between selected health problems and exposures among women semiconductor workers in Malaysia. AB - A study conducted between 1998-2001 on the semiconductor industry in Penang and Selangor found that irregular menstruation, dysmenorrhea and stress were identified as the three leading health problems by women workers from a checklist of 16 health problems. After adjusting for confounding factors, including age, working duration in current factory, and marital status, in a multiple logistic regression model, wafer polishing workers were found to experience significantly higher odds of experiencing irregular menstruation. Dysmenorrhea was found to be significantly associated with chemical usage and poor ventilation, while stress was found to be related to poor ventilation, noise and low temperatures. PMID- 14750380 TI - Core needle biopsy of palpable breast lump: the influence of needle size. AB - The diagnostic value of core needle biopsy is increasingly being preferred because of its better characterization of benign and malignant lesions and lower frequency of insufficient samples. The aim of this study was to determine the diagnostic accuracy and complication rates with 2 different gauges of core biopsy needle in the preoperative diagnosis of palpable breast lumps. A total of 150 consecutive core biopsies were included in this prospective non-randomised study of palpable breast lump from May 2000 to May 2001. The tissue diagnosis made from the core biopsy specimen was compared with the final histopathology reports from the excised specimen. However, if the lump is not excised, a presumptive diagnosis of benign lesion was made only after at least 6 months follow up with no change in the breast lump. The data were analysed for sensitivity, specificity, predictive values, diagnostic accuracy and complications. The results from the 2 different sizes of core needle biopsies were compared accordingly and a statistical analysis was performed using Chi-squared test. Ninety-six core specimens were acquired with 14 G needle while the other 54 with 16 G needle. There was no significant statistical difference between the accuracy of both needle sizes. However, 4 complications occurred with the larger size 14 G needle while none with the 16 G needle, but this was not statistically significant. In conclusion the size 16 G core biopsy needle provided an accurate diagnostic reliability that is comparable to the larger size 14 G needle in the preoperative diagnosis of palpable breast lump. PMID- 14750381 TI - Open-globe injuries: the experience at Hospital Sultanah Aminah, Johor Bahru. AB - Between 1st January 1999 and 31st December 2000, 152 patients (156 eyes) with open-globe injuries were treated in the Department of Ophthalmology, Hospital Sultanah Aminah, Johor Bahru. The majority were male (88.2%), Malay (63.2%), from the Johor Bahru district (51.3%) and aged between 21 and 30 years (23.7%). Most injuries were workplace-related (41.4%). Lens injury, retinal detachment, endophthalmitis, intraocular foreign bodies and phthisis occurred in 40.4%, 15.4%, 14.7%, 12.2% and 11.5% of eyes respectively. A favourable visual outcome occurred in 55.4% of eyes. Prognostic factors for visual outcome include presenting visual acuity, relative afferent pupillary defect, wound location, lens injury, retinal detachment and endophthalmitis. PMID- 14750382 TI - Perioperative death in Malaysia: the transition phase from a developing nation to a developed one. AB - This paper examines the surgical pathology associated with perioperative deaths in a country that is undergoing the transition from a developing to a developed nation status. The data from an ongoing nation-wide perioperative mortality study was prospectively collected for the period July 1996 to December 1997 and analyzed. The surgical pathology related to perioperative deaths in Malaysia is different from other developing and developed countries. While death from trauma and the late presentation of surgical conditions are similar to developing countries, infective gastrointestinal conditions were rarely encountered. Diseases associated with advanced age such as colorectal cancer, peptic ulcer, urological diseases and vascular conditions are beginning to emerge. As the country races towards a developed nation status, increasing life expectancy and changing life-styles are expected to influence the disease pattern. The planning of surgical facilities and manpower development must recognize the changes taking place. PMID- 14750383 TI - The outcome of abdominal aortic aneurysm repair in northern Malaysia. AB - A prospective study of all infrarenal abdominal aortic aneurysm (AAA) repairs both as electives and emergencies in Penang between January 1997 to December 2000 is presented. The objectives of the study were to determine the age, gender, racial distribution of the patients, the incidence, and risk factors and to summarize treatments undertaken and discuss the outcome. Among the races, the Malays were the most common presenting with infrarenal AAA. The mean age of patients operated was 68.5 years. Males were more commonly affected compared to females (12:1). Most infrarenal AAA repairs were performed as emergency operations, 33 cases (61.1%) compared to electives, 21 cases (38.9%). Total survival was 70.3% (elective 85.7%; emergency 57.6%). Mortality rate was 31.5% and the primary reason is the lack of operating time available for urgent operation and for treatment of concurrent disease states. Mycotic aneurysm with its triad of abdominal pain, fever and abdominal mass resulted in a significantly higher mortality (46.6%). Ninety six percent of the infrarenal AAA had transverse diameter greater than 6 cm. Morphologically 90.7% were fusiform AAA rather than saccular aneurysm (9.3%). Pulmonary complications (35.2%) were more common than cardiac complications (11.1%) possibly related to the urgent nature of the operation, smoking or history of pulmonary tuberculosis. Bleeding (14.8%) was the most common cause of mortality in ruptured mycotic infrarenal AAA. PMID- 14750384 TI - A study of 415 cases of esophageal carcinoma in northwest of Iran. AB - BACKGROUND AND OBJECTIVE: Considering the serious and fatal nature of esophageal carcinoma and as prevalence in the population on the banks of Caspian sea and northern region (Azerbaijan, Khorassan, Gilan, Mazandaran, Golestan and Kurdistan) along with the importance of its early diagnosis in the initial stage in order to increase the survival period of the patient, we aimed to proceed in regard to these factual observations so that this study should be an incentive and beginning of our future perfect study. MATERIALS AND METHODS: In a prospective study over a period of five years (from year 1994 to 1999), a total of 415 pathologically documented esophageal carcinoma out of those 33000 referred cases those with clinical signs and symptoms of esophageal cancer were enrolled in this study. Main complain of patients included (Dysphagia, odynophagia retrosternal pain with unknown origin, anorexia, weight loosE, and anemia with undetermined etiology) in one of the treatment center Age of these patients ranged from 20 to 80 years old, with mean age of 60 years and stable variables were selected from the chart of the patients. The study was accomplished with serial radiography and Endoscopy. Biopsy specimens collected during Endoscopy were sent for histological examination and staging of the disease. Finally, with the confirmation of pathological report, patients were referred for the therapeutical management. RESULTS: Out of 415 patients with distinct confirmative pathological findings, 349 (88.3%) cases had squamous cell carcinoma while, 46 (11.7%) of them were having adenocarcinoma. Among these patients, 214 were males and 201 females. The male to female ratio was almost equal. CONCLUSION: This study highlights the type of esophageal cancer along with the sex distribution and its diagnostic approaches. Squamous cell carcinoma was mainly observed among the patients studied and there was not significant difference between its involvement among males and females. After the diagnosis, 5 years survival rate will depend upon its early diagnosis, accurate staging of the disease and management of the patient with appropriate therapy. PMID- 14750385 TI - Surgical intervention in traumatic facial nerve paralysis. AB - A four years review from June 1998 to June 2002 of traumatic facial nerve paralysis from temporal bone fractures that required surgical intervention is presented. The aim of this clinical presentation was to determine the current pattern of cases with traumatic facial paralysis which required surgical intervention at our center. There were six cases, of which four (66%) were longitudinal fractures, one each (17%) had transverse fracture and fracture over the lateral wall of mastoid. Hearing loss (83%) was the commonest associated clinical symptom. All cases underwent decompression via the transmastoid surgical approach. Intraoperative findings revealed oedema of facial nerve involving vertical segment and horizontal segment in three cases each respectively. Two cases had concomitant bony impingement. The facial nerve functions in four cases (66%) and one case recovered to House Brackmann grade 2 and 4, 12 months and 3 months respectively postsurgery. The case with transverse fracture remained as House Brackmann grade 5 after two years. PMID- 14750386 TI - The Jarisch-Herxheimer reaction in ocular syphilis. AB - A patient with ocular syphilis is presented. She experienced deterioration in vision following the commencement of treatment due to a Jarisch-Herxheimer reaction. This is a transient febrile illness that can occur in patients after the first adequate dose of an anti-microbial drug to treat infectious diseases such as syphilis, Lyme disease and relapsing fever. However, a Jarisch-Herxheimer reaction occurring in a patient receiving treatment for ocular syphilis can be serious, resulting in the rapid loss of vision. PMID- 14750387 TI - Double pylorus. AB - Double pylorus (DP) or duplication of the pylorus is an uncommon condition that is either congenital or acquired. Acquired double pylorus (DP) results from a peptic ulcer eroding through and creating a fistula between the duodenal bulb and the distal stomach. We report a case of an acquired double pylorus in an adult gentleman that resulted from the erosion of a duodenal and prepyloric ulcer. PMID- 14750388 TI - Combined needlescopic cholecystectomy and laparoscopic splenectomy for the treatment of thalassaemic splenomegaly and cholelithiasis. AB - Gallstone disease is a common association in patients with haematological splenomegaly. When indicated, simultaneous splenectomy and cholecystectomy should be performed and traditionally this is accomplished by open surgery. We report a 17 year old thalassaemic girl with splenomegaly complicated by gallstone pancreatitis. We treated her with a combination of needlescopic cholecystectomy and laparoscopic splenectomy as well as delivering the huge spleen via a pfannenstiel incision to hide the scar. We believe this technique is an acceptable alternative mainly for rapid delivery of the spleen and to minimize visible scars hence improving cosmesis. PMID- 14750389 TI - Primary aorto-duodenal fistula. AB - Primary aorto-duodenal fistula is a rare and life-threatening cause of upper gastro-intestinal bleed. In this case report, a patient presented acutely with several episodes of haematochezia and pulseless lower limbs bilaterally. Primary aorto-duodenal fistula with peripheral vascular disease was diagnosed after an urgent CT angiogram was performed. She underwent left axillo-bifemoral bypass, resection of the fistula, Rouxen-Y gastro-jejunostomy, pyloric exclusion and controlled duodenal fistula the following day. PMID- 14750390 TI - The different faces of facial nerve schwannomas. AB - Facial nerve schwannomas are rare benign tumors. The tumor can arise anywhere along the course of the facial nerve. The most common presentation for this tumor is a slowly progressive facial nerve paralysis. Sensorineural hearing loss (SNHL) and tinnitus are later symptoms. The symptoms and signs depend on the site of tumor along the nerve. We report three cases of facial nerve schwannomas with different clinical presentations. Appropriate management of a facial nerve schwannoma should be based on the site and extent of the tumor and status of the nerve function. PMID- 14750391 TI - Evaluation of three commercial rapid tests for detecting antibodies to human immunodeficiency virus. AB - Determine HIV-1/2, Chembio HIV-1/2 STAT-PAK and PenTest are simple/rapid tests for the detection of antibodies to HIV-1 and HIV-2 in human whole blood, serum and plasma samples. The assay is one step and the result is read visually within 15 minutes. Using 92 known HIV-1 reactive sera and 108 known HIV-1 negative sera, the 3 HIV tests correctly identified all the known HIV-1 reactive and negative samples. The results indicated that Determine HIV-1/2, Chembio HIV-1/2 STAT-PAK and PenTest HIV are as sensitive and specific (100% concordance) as Microparticle Enzyme Immunoassay. The data indicated that these 3 HIV tests are effective testing systems for diagnosis of HIV infection in a situation when the conventional Enzyme Immunoassay is not suitable. PMID- 14750392 TI - Immunohistochemical localization of RET rearrangements in a Malaysian papillary thyroid carcinoma population. PMID- 14750393 TI - Reevaluating the use of antibiotics in acute otitis media in children. AB - Increasing concern about the antibiotic resistance in acute otitis media (AOM) has led to debate over use of antibiotic in AOM and duration of therapy. Many studies have proved that watchful waiting should be used more often for acute otitis media. In children over two years, the most appropriate treatment was found to be initial observation followed by 5 days of an antibiotic if the child failed to improve spontaneously. In children less than 2 years or one with severe symptoms antibiotic can be started after 24 hours if there is no improvement with symptomatic treatment. Physician should be more selective in the prescription of antibiotics early in AOM. PMID- 14750395 TI - Imperfect Medicare law has positive implications for pharmacists. PMID- 14750396 TI - WHO, UNAIDS set HIV treatment goal for developing countries. Plan is to treat 3 million with HIV infection by 2005. PMID- 14750394 TI - An introduction to the Medicare Prescription Drug, Improvement, and Modernization Act of 2003. PMID- 14750397 TI - Protocol sharing requires diligence by recipients. PMID- 14750398 TI - National specialty pharmacies focus on rare, chronic diseases. PMID- 14750399 TI - The pharmaceutical supply chain: a perfect storm is brewing. PMID- 14750400 TI - Projecting future drug expenditures--2004. AB - PURPOSE: Drug expenditure trends since 2002 and projected drug expenditures for 2004 are discussed. SUMMARY: In 2002 there was a moderation in the trend of increasing drug expenditures. Drug expenditures increased by 12.3% between 2001 and 2002. This trend continued in the first half of 2003, with expenditures increasing by only 10% compared with 2002. This moderation in the drug expenditures trend can be attributed to many factors, especially patent expirations and decreases in new drug approvals. Higher cost sharing for consumers and a general economic slowdown in the United States affecting employment and insurance coverage have resulted in a smaller increase in drug utilization. In 2004, there should be a 10-12% increase in drug expenditures for outpatient settings, a 19-21% increase for clinics, and a 6-8% increase for hospitals. CONCLUSION: Drug expenditure growth should continue to outpace the growth in overall health care expenditures and the growth in the U.S. economy. PMID- 14750401 TI - Diabetic neuropathy: an intensive review. AB - PURPOSE: The epidemiology, classification, pathology, and treatment of diabetic neuropathy are reviewed. SUMMARY: Diabetic peripheral neuropathy is a common complication of diabetes that can cause significant morbidity and mortality. Some 30% of hospitalized and 20% of community-dwelling diabetes patients have peripheral neuropathy; the annual incidence rate is approximately 2%. The primary risk factor is hyperglycemia. Sensorimotor neuropathy is marked by pain, paresthesia, and sensory loss. Cardiac autonomic neuropathy (CAN) may contribute to myocardial infarction, malignant arrhythmia, and sudden death. Gastroparesis is the most debilitating complication of gastrointestinal autonomic neuropathy. Genitourinary autonomic neuropathy can cause sexual dysfunction and neurogenic bladder. The pathology of diabetic neuropathy involves oxidative stress, advanced glycation end products, polyol pathway flux, and protein kinase C activation; all contribute to microvascular disease and nerve dysfunction. For symptom management current evidence from clinical trials supports the use of desipramine, amitriptyline, capsaicin, tramadol, gabapentin, bupropion, and venlafaxine as preferred medications. Citalopram, nonsteroidal antiinflammatory drugs, and opioid analgesics may be used as adjuvant agents. Lamotrigine, oxcarbazepine, paroxetine, levodopa, and alpha-lipoic acid are alternative considerations. Evidence supporting the use of zonisamide, fluoxetine, mexiletine, dextromethorphan, and phenytoin is considered equivocal. Complementary therapies have also shown efficacy. The symptoms of CAN may be ameliorated with fludrocortisone, clonidine, midodrine, dihydroergotamine or caffeine, octreotide, and beta-blockers. Gastroparesis may be treated with metoclopramide or erythromycin. The most promising disease-modifying therapy is ruboxistaurin, which is in Phase III trials. Glycemic control remains the foundation of prevention and the prerequisite of adequate treatment. CONCLUSION: Diabetic neuropathy is a many-faceted complication of diabetes that can be managed symptomatically with an array of drugs. PMID- 14750402 TI - Preventing medication errors with smart infusion technology. AB - PURPOSE: Processes that pharmacists can use to identify high-risk areas and drugs that require special focus in error-prevention efforts are discussed, with emphasis on the need for i.v. medication error prevention. SUMMARY: Pharmacists can help determine where best to focus medication safety efforts and innovative technology by identifying areas that pose the greatest risk of harm to a patient, such as medications, administration routes, patient care areas, and diagnosis related groups. Delivery of i.v. medications via infusion devices has traditionally not been a major concern for pharmacists. The introduction of "smart" infusion technology has changed that paradigm by requiring pharmacist involvement in defining minimum and maximum doses for continuous and bolus infusions used within a health care facility. This technology provides a software filter to prevent key-stroke errors in programming infusion devices for delivery of i.v. drugs, as well as a new source of data with which to measure medication errors at the bedside. Implementation of smart infusion technology at Vanderbilt University Medical Center appeared to prevent errors involving heparin. In addition to having an immediate positive impact at the bedside, the technology was relatively easy to implement. CONCLUSION: Smart infusion systems represent an innovative technology that can provide an additional layer of protection at the point of care to help avert i.v. drug errors and prevent patient harm. PMID- 14750403 TI - Association between hospital size and quality improvement for pharmaceutical services. AB - PURPOSE: The relationship between hospital size and quality improvement (QI) for pharmaceutical services was studied. METHODS: A questionnaire on QI was sent to hospital pharmacy directors in Michigan and Florida in 2002. The questionnaire included items on QI lead-team composition, QI tools, QI training, and QI culture. RESULTS: Usable responses were received from 162 (57%) of 282 pharmacy directors. Pharmacy QI lead teams were present in 57% of institutions, with larger teams in large hospitals (> or = 300 patients). Only two QI tools were used by a majority of hospitals: root-cause analysis (62%) and flow charts (66%). Small hospitals (< 50 patients) were less likely than medium-sized hospitals (50 299 patients) and large hospitals to use several QI tools, including control charts, cause-and-effect diagrams, root-cause analysis, flow charts, and histograms. Large hospitals were more likely than small and medium-sized hospitals to use root-cause analysis and control charts. There was no relationship between hospital size and the frequency with which physician or patient satisfaction with pharmaceutical services was measured. There were no differences in QI training or QI culture across hospital size categories. CONCLUSION: A survey suggested that a majority of hospital pharmacies in Michigan and Florida have begun to adopt QI techniques but that most are not using rigorous QI tools. Pharmacies in large hospitals had more QI lead-team members and were more likely to use certain QI tools, but there was no relationship between hospital size and satisfaction measurements, QI training, or QI culture. PMID- 14750404 TI - Computerized prescriber order-entry systems: evaluation, selection, and implementation. PMID- 14750405 TI - Using an infection-reporting database to improve outcomes in cataract-surgery patients. PMID- 14750406 TI - Criteria for use of drotrecogin alfa. PMID- 14750407 TI - Criteria for use of drotrecogin alfa. PMID- 14750409 TI - [Does cardiac resynchronization therapy improve nitric oxide concentration in exhaled gas?]. AB - OBJECTIVES: Cardiac resynchronization therapy (CRT) is widely known to improve cardiac function in patients with chronic heart failure, especially those with increased intraventricular conduction delay. However, whether vascular endothelial cell function is improved remains unknown. Capability of nitric oxide (NO) production is a good marker to evaluate endothelial cell function. Therefore, NO output was measured in exhaled air in patients with chronic heart failure after CRT. METHODS: Six patients with chronic heart failure and dilated cardiomyopathy (mean age 62.0 +/- 11.3 years, left ventricular ejection fraction 22.8 +/- 11.6%) received CRT. Pacing leads were lodged at the right atrium, the apex of the right ventricle and at the mid portion of the postero-lateral branch of the coronary sinus. One month after CRT, endtidal NO concentration was evaluated with and without pacing using the chemiluminescence method. RESULTS: Left ventricular ejection fraction improved from 22.8% to 28.8% after the procedure. Endtidal NO was significantly (p = 0.038) increased from 64.7-13.2 ppb without pacing to 69.1-14.1 ppb with pacing. Respiratory rate (with pacing: 19.7 5.7 n/min, without pacing: 20.3-4.8 n/min), tidal volume (with pacing: 523.2 122.7 ml, without pacing: 516.2-160.3 ml) and minute ventilation were not changed. CONCLUSIONS: CRT did not affect respiratory rate, tidal volume or minute ventilation, so increased endtidal NO did not result from enhanced production from tracheal and/or bronchial epithelial cells. Therefore, endtidal NO increased because chronic heart failure increased pulmonary perfusion and so raised vascular shear stress. CRT for patients with dilated cardiomyopathy can improve vascular endothelial cell function. PMID- 14750408 TI - [Detection of myocardial ischemia by analysis of the shape of exercise-induced ST elevation in infarct-related leads]. AB - OBJECTIVES: Possible mechanisms of exercise-induced ST elevation in infarct related leads include ventricular dyskinesis, and myocardial ischemia in the infarct region. Detection of ischemia in viable myocardium in the infarct region is important to determine the therapeutic strategy. This study evaluated whether the analysis of the shape of exercise-induced ST elevation(convex or concave type) is useful to detect myocardial ischemia in the infarct region. METHODS: Ninety-eight patients (78 males, 20 females, mean age 59 +/- 10 years) with prior Q wave myocardial infarction underwent the treadmill exercise test. Patients were divided into three groups according to the exercise-induced ST changes: No ST-E group, 27 patients without ST changes; Concave ST-E group, 52 patients with concave type ST elevation; Convex ST-E group, 19 patients with convex type ST elevation. Coronary arteriography was evaluated in all patients. Dobutamine stress echocardiography was performed in 38 patients, including 28 patients in the Concave ST-E group and 10 patients in the Convex ST-E group. Biphasic or worsening response on dobutamine stress echocardiography was defined as ischemic response. RESULTS: Coronary arteriography revealed significant stenosis of the infarct-related artery in 30% of the No ST-E group, 47% in the Convex ST-E and 86% in the Concave ST-E groups (p < 0.05). Dobutamine stress echocardiography revealed myocardial ischemia in the infarct region in 30% in the Convex ST-E group and 75% in the Concave ST-E group(p < 0.05). CONCLUSIONS: The Concave ST-E group had a higher incidence of stenosis of the infarct-related artery and myocardial ischemia in the infarct region. Analysis of the shape of exercise induced ST elevation in infarct-related leads is useful for the detection of ischemia of viable myocardium. PMID- 14750410 TI - [Usefulness of transthoracic freehand three-dimensional echocardiography for the evaluation of mitral valve prolapse]. AB - BACKGROUND: Two-dimensional (2D) echocardiography is routinely used in evaluating patients with mitral valve prolapse but requires a systematic examination for accurate assessment of the involved lesion of mitral valve prolapse, because the sonographer is required to mentally reconstruct two-dimensional images into three dimensions. Recently, freehand three-dimensional (3D) echocardiography has been introduced in the clinical setting for three-dimensional visualization of the mitral valve apparatus. OBJECTIVES: To evaluate the accuracy of the freehand 3D echocardiography system in assessing the involved lesion in patients with mitral valve prolapse. METHODS: This study consisted of 25 consecutive patients (15 men, 10 women, mean age 55 +/- 17 years) with mitral valve prolapse who were scheduled for 3D echocardiography. Mitral valve was reconstructed in the view from the left atrium (surgeon's view) by 3D echocardiography. The location of the involved lesion in mitral valve was classified as the medial, middle and lateral portions of the anterior leaflet, and the medial, middle and lateral scallops of the posterior leaflet, respectively. The results by 3D echocardiography were compared with those of 2D echocardiography as the clinical standard. RESULTS: An adequate three-dimensional display of the entire mitral valve for analysis of the involved lesion could be reconstructed in all 25 patients (feasibility 100%). The sensitivity of 3D echocardiography for detecting the lesions at the medial, middle and lateral portions of the anterior leaflet was 80%, 100% and 75%, and the medial, middle and lateral scallops of the posterior leaflet was 100%, 100% and 0%, respectively. The specificities were 100% at all locations in the mitral valve. CONCLUSIONS: These results indicate that freehand 3D echocardiography is useful for assessment of the involved lesion of the mitral valve in patients with mitral valve prolapse. PMID- 14750411 TI - Experimental study of quantitative assessment of left ventricular mass with contrast enhanced real-time three-dimensional echocardiography. AB - OBJECTIVES: To evaluate the feasibility and accuracy of measurement of left ventricular mass with intravenous contrast enhanced real-time three-dimensional (RT3D) echocardiography in the experimental setting. METHODS: RT3D echocardiography was performed in 13 open-chest mongrel dogs before and after intravenous infusion of a perfluorocarbon contrast agent. Left ventricular myocardium volume was measured according to the apical four-plane method provided by TomTec 4D cardio-View RT1.0 software, then the left ventricular mass was calculated as the myocardial volume multiplied by the relative density of myocardium. Correlative analysis and paired t-test were performed between left ventricular mass obtained from RT3D echocardiography and the anatomic measurements. RESULTS: Anatomic measurement of total left ventricular mass was 55.6 +/- 9.3 g, whereas RT3D echocardiographic calculation of left ventricular mass before and after intravenous perfluorocarbon contrast agent was 57.5 +/- 11.4 and 55.5 +/- 9.3 g, respectively. A significant correlation was observed between the RT3D echocardiographic estimates of total left ventricular mass and the corresponding anatomic measurements (r = 0.95). A strong correlation was found between RT3D echocardiographic estimates of left ventricular mass with perfluorocarbon contrast and the anatomic results (r = 0.99). Analysis of intraobserver and interobserver variability showed strong indexes of agreement in the measurement of left ventricular mass with pre and post-contrast RT3D echocardiography. CONCLUSIONS: Measurements of left ventricular mass derived from RT3D echocardiography with and without intravenous contrast showed a significant correlation with the anatomic results. Contrast enhanced RT3D echocardiography permitted better visualization of the endocardial border, which would provide a more accurate and reliable means of determining left ventricular myocardial mass in the experimental setting. PMID- 14750412 TI - [Acute myocardial infarction due to septic emboli in a patient with severe pneumonia: a case report]. AB - A 77-year-old man presented with acute anterior myocardial infarction and cardiogenic shock. We successfully performed coronary angioplasty and stenting with intra-aortic balloon pumping for a proximal lesion of the left anterior descending artery, which supplied the territory of the totally occluded right coronary artery. He also had pneumonia resistant to antibiotic therapy. On the 11th day after the onset of infarction, he suffered cardiogenic shock again and died. Postmortem histological examination demonstrated multiple septic emboli with neutrophil infiltration, resulting in myocardial infarctions. There was no occlusion at the stenting site. Severe pneumonia was found in both lungs, which may have been the embolic source in this case. PMID- 14750413 TI - [A 55-year-old man with prior myocardial infarction presenting with congestive heart failure refractory to medical treatment]. PMID- 14750415 TI - The microstructural border between the motor and the cognitive domain in the human cerebral cortex. AB - When we voluntarily interact with our environment, the agranular frontal cortex (Brodmann's areas 4 and 6) plays a pivotal role in cortical motor control. The primary motor cortex (area 4) influences kinematic and dynamic parameters of movements, whereas the rostrally adjoining nonprimary motor cortex (area 6) uses external (e.g., sensory) or internal cues to trigger and guide movements. Once thought to be homogeneous, data from nonhuman primates have shown that area 6 is a mosaic of areas, each with distinct structural and functional properties: the supplementary motor areas "SMA proper" and "pre-SMA" on the mesial cortical surface, and the dorso- and ventrolateral premotor cortex on the cortical convexity. Dorso- and ventrolateral premotor areas are specifically connected with posterior parietal areas. These parieto-frontal circuits work in parallel and tranform different aspects of sensory information into appropriate motor commands. The rostral border of area 6 is very important for functional neuroimaging studies in humans since it separates the "motor domain" of the supplementary motor/premotor cortex from the "cognitive domain" of the prefrontal cortex. Can the topography of this border be inferred from the gyral pattern of the frontal lobe? To answer this, ten postmorterm brains were scanned with a T1 weighted magnetic resonance sequence. The brains were serially sectioned at 20 micro M and area 6 was defined by subjective and objective cytoarchitectonic analysis. Each brain's histological volume (with the representation of area 6) was reconstructed in 3-D and spatially normalized to the reference brain of a computerized atlas. The ten normalized volumes were superimposed and a population map was generated that describes, for each voxel, how many brains have a representation of area 6. On the mesial coetical surface, the rostral border of area 6 lies rostral to the anterior commissure-- though the distance varies across different brains. On the lateral convexity, the border recedes in a caudal direction-- again to a varying degree in different brains-- and lies on the precentral gyrus close to the sylvian fissure. No macroanatomical landmark indicates the border between area 6 and the prefrontal cortex. The question whether a motor task engages only the "motor domain" of the supplementary motor/premotor cortex or in addition the "cognitive domain" of the prefrontal cortex can only be answered by superimposing the functional activation map with the microstructural population map of area 6. PMID- 14750416 TI - [Assessment of serum levels of adhesion molecules (sICAM-1, sVCAM-1, sE-selectin) in stable and unstable angina and acute myocardial infarction]. AB - The targeting and recruitment of inflammatory cells to vascular endothelium in ischaemic heart diseases is mediated by Intercellular Adhesion Molecule-1 (ICAM 1), Vascular Cell Adhesion Molecule (VCAM-1) and E-selectin and proinflammatory cytokines. Accumulation of mononuclear cells to the endothelium is one of the earliest events in the formation of an atherosclerotic lesion. The aim of this study was to estimate the serum concentrations of soluble intercellular adhesion molecule-1 (sICAM-1), vascular adhesion molecule-1 (sVCAM-1), selectin E (sE selectin) in patients with acute myocardial infarction (Group 1--n = 18 patients: 3 women and 15 men, mean age--60 years), unstable angina pectoris (Grupa 2--n = 31 patients: 8 women and 23 men, mean age--62 years and stable heart disease (Grupa 3--n = 25 patients: 14 women and 11 men, mean age--61 years. The control group (Group 4--n = 20) consist of twenty healthy patient without coronary risk factors. ELISA method was used to determine the concentration of adhesion molecules of acute inflammation parameters, and traditional risk factors with using standard methods. The serum levels of sICAM-1, sVCAM-1 were markedly elevated in patients with acute myocardial infarction, unstable angina pectoris and stable heart disease compared to control group (p < 0.001, p < 0.004, p < 0.0002 for sICAM-1, p < 0.007, p < 0.003, p < 0.004 for sVCAM-1). Serum concentration of sE-selectin in three groups was similar, we did not find statistically significant differences between them. Furthermore, serum concentrations of adhesion molecules correlated with serum concentrations of acute inflammation parameters and traditional coronary risk factor for example BMI, systolic and diastolic blood pressure and lipid concentration. Additional serum concentration of sICAM-1 was elevated in smoking patients compared to non smokers. We conclude that evaluation of adhesion molecules (sICAM-1 and sVCAM-1 in patients with heart diseases can be unspecific markers of activity of inflammatory process in coronary vascular endothelium. PMID- 14750417 TI - [Coronary risk factors and mode of ischemic heart disease treatment in patients over 65 years of age]. AB - The aim of the study was to investigate the presence of coronary risk factors, the frequency of coronary angiography and methods of treatment analysis in patients over 65 years old. MATERIAL: Among 670 patients hospitalized in our institution, 136 were in the age over 65 years old (20.3%) and they were the study group, 534 patients were in the age below 65 years old and they were the control group. METHODS: Total cholesterol, HDL cholesterol, triglycerides, plasminogen inhibitor plasminogen type 1 (PAI-1) and fibrinogen were measured, whereas LDL cholesterol was calculated by means of Friedewald's formula. The frequency of such coronary risk factors as: smoking, arterial hypertension, diabetes and family history of coronary artery disease (CAD), previous myocardial infarction (MI), arterial hypertension and diabetes were analysed. Also were studied results of coronary angiography and qualification for different mode of treatment: pharmacological therapy, percutaneous transluminal coronary angioplasty or coronary artery bypass grafting. STATISTICS: Chi-square test or t Student test were used. RESULTS: In the group of patients over 65 years old there were more females. The frequency of arterial hypertension and fibrinogen level were higher in the older group than in the younger one. The frequency of smoking (current and in the past) was higher in the younger patients. The only difference in lipid parameters was significantly higher triglycerides level in the younger group. The percent of people with treated hypercholesterolemia was higher in the younger patients. There were no significant difference in frequency of the other coronary risk factors in both groups. Analysis of family history revealed the lower frequency of stroke, CAD and MI in older patients' relatives. Q wave MI was diagnosed in 32.4% of older patients and in 36.2% of younger ones. Non-Q wave MI was noted in 25.7% and 26.1% respectively. In patients over 65 years old the frequency of unstable CAD was significantly higher than in the younger group. In coronary angiography one vessel changes stenosis was diagnosed in 11.5% of older patients, whereas three vessels stenosis in 40% of these people. Although in both groups invasion treatment was instituted in 60% of the patients, the frequency of coronary surgery was significantly higher in older patients (34.2% vs 22.5%; p < 0.01) whereas the frequency of angioplasty was lower in patients over 65 years old (25.5% vs 44.3%; p < 0.005). CONCLUSIONS: 1. In patients over 65 years old the frequency of coronary risk factors was lower than in younger ones. All these factors, except arterial hypertension, seem not to be of such great importance as in younger people. 2. The age alone should not restrict instituting invasion treatment, by which were treated 60% of patients in both groups. 3. In opposition to the control group, in the patients over 65 years old prevailing method of invasion treatment was coronary surgery. PMID- 14750418 TI - [Results of coronary heart disease treatment with coronary angioplasty in patient over 65]. AB - The aim of the study was to investigate the results of coronary angioplasty in patients over 65 years old. MATERIAL: Among the following 590 patients after coronary angiography performed in the Department of Cardiology, Institute of Cardiology Medical University of Lodz 113 (19.2%) pt were over 65 years old (mean age 69.4 years) and 477 (80.8%) pt in the age below 65 years old (mean age 51.6 years). Coronary angioplasty underwent 31 patients (27.4%) over 65 years old and which were the study group and 207 (43.4%) pt below 65 years old and which were the control group. METHODS: The frequency of previous MI, CCS angina class, the results of coronary angiography and qualification for percutaneous transluminal coronary angioplasty treatment, the percent of successful and complete revascularisation, the number of clinical restenosis and restenosis in angiography and the methods of restenosis treatment were estimated. In statistical analysis Chi-square test and t-Student test were used. RESULTS: Three vessels disease in angiography was observed more frequently in pt over 65 years old than in the younger group (40.7% vs 26.6%; p < 0.005) and these pt were qualified more frequently to surgery revascularisation 37.2% (vs 22% in younger ones; p < 0.001) while the coronary angioplasty was performed only in 27.4% pt (vs 43.4% in younger ones; p < 0.005). Successful angioplasty was performed in almost 97 percent of patients in both groups. The frequency of complete revascularization was significantly lower in patients over 65 years old than in the control group (36.7% vs 57.9%, p < 0.05) because of the more advanced coronary lesions in older ones. Angioplasty with stent implantation was performed in 29% pt over 65 and in 27% pt below 65 years old. On the basis of clinical evidence of ischemia (mainly recurrent angina) restenosis was suspected in 36.7% of older and in 29.7% of younger patients but the difference wasn't statistically significant. Angiographic analysis revealed restenosis after coronary balloon angioplasty in 33.3% patients over 65 years old and in 21.3% of younger ones. Stenosis in stents was observed in 13.3% of older and in 6.4% of younger patients. Restenosis was treated with coronary artery bypass grafting in about 20% and with percutaneous revascularization in 80% of patients in both groups. In patients over 65 years old the frequency of using intracoronary stents during revascularization was lower than in the control group (10% vs 30%). CONCLUSIONS: 1. In patients over 65 years old the frequency of using angioplasty as the method of revascularization was lower than in younger patients. 2. Although complete revascularization can be instituted only in half of the patients over 65 years old, the results of coronary angioplasty are not worse than in younger people. PMID- 14750419 TI - [Met-enkephalin plasma concentration and content in liver tissue in patients with primary biliary cirrhosis]. AB - Met-enkephalin is a pentapeptide belonging to the opioid system and like other opioid peptides is involved in phenomena associated with modulated pain perception, regulation of memory and emotional conditions, food and liquid consumption, regulation of immunological system and it has an impact upon digestive system motility, gastric and pancreatic secretion, metabolism of carbohydrates. Met-enkephalin concentration changes in liver tissue and plasma during liver disorders. It has been revealed that Met-enkephalin takes part in the etiopathogenesis of ascites and pruritus. Primary biliary cirrhosis (PBC) is a chronic inflammatory liver disease, its course involves the destruction of small and medium-sized bile ducts as a result of the process of granulation tissue growth developing in such diseases as fibrosis, liver cirrhosis development, and liver insufficiency. During the ending period of the PBC, clinical and biochemical symptoms of decompensation of liver cirrhosis occur. The objective of the study and paper was to make a comparison between the Met enkephalin concentration rates in plasma and its content in liver tissue with the progress stage of PBC, which was assessed using clinical symptoms. The study was carried out in 40 female patients with PBC. PBC has been found by means of biochemical, serological investigations as well as histopathological liver bioptate estimation. The patietns with different causes of liver disorders have been excluded. Met-enkephalin concentration in plasma and content in liver tissue have been indicated in each patient. The control group consisted of 30 female patients treated in the Department of Gastroenterology with normal activity of indicative and cholestatic enzymes, and without any organic disorders. In the control group, the Met-enkephalin concentration rates in plasma were determined. Statistically significant difference has been indicated in Met-enkephalin plasma concentration between two patients groups: 23 patients with pruritus and 17 patients without it where (p = 0.0476). Statistically significant difference in Met-enkephalin concentration level in plasma and liver tissue in the group of 19 patients with liver failure symptoms (stadium IV and V according to Schaffner and Popper classification) and in the group of 21 patients without those symptoms (stadium I and II according to Schaffner and Popper classification) has been stated. For Met-enkephalin plasma concentration p = 0.002 and for Met-enkephalin liver tissue concentration p = 0.0214. CONCLUSIONS: 1. The Met-enkephalin concentration rates in serum of patients with the PBC is higher than in the control group; 2. In PBC patients' pruritus appears together with the increase of Met-enkephalin plasma concentration. 3. In PBC patients, when liver failure symptoms appear Met-enkephalin concentration increases in plasma, and it decreases in the liver tissue. PMID- 14750420 TI - [Efficacy and feasibility evaluation in long term pamidronate treatment of bone metastases]. AB - The aim of our study was to evaluate the efficacy and feasibility of long-term pamidronate treatment. MATERIAL AND METHODS: Thirty-six patients (pts) undergoing long-term (> 9 months) pamidronate treatment for bone metastases of breast cancer (30 pts), prostate cancer (3), multiple myeloma (2) and renal carcinoma (1) were retrospectively analyzed. The indication for pamidronate treatment were appearance of bone metastases (21 pts), progression of bone lesions (13) or intolerance of clodronate (2). Pamidronate was administered as an intravenous infusion, most commonly at a dose of 90 mg monthly. Skeletal complications including pathologic fractures, the need for palliative radiotherapy or bone surgery, spinal cord compression and hypercalcemia as well as occurrence of new bone or visceral lesions were assessed. The use of analgesics and subjective bone pain relief were used to evaluate the analgetic effect of pamidronate therapy. Adverse events of pamidronate treatment were noted. RESULTS: Patients received a median of 15 pamidronate infusions (range 9-35). Complete pain control was observed in 7 pts (19%), partial in 21 (58%) and stabilization in 8 (22%). Mean time to maximal effect was 5 months (range 0-17). There were 5 cases (14%) of fever and 6 cases (17%) of flu-like syndrome after pamidronate administration. New bone lesions appeared in 16 pts (44%) after a median of 12 months (range 1 28). Other skeletal complications included pathologic fractures (9 pts, 25%) and hypercalcemia (2 pts, 5.6%); 13 pts (36%) required radiotherapy. Symptomatic progression occurred in 27 pts (75%), with a median progression-free time of 14 months (range 5-35) from the beginning of pamidronate treatment. CONCLUSIONS: Long-term treatment with pamidronate in patients with bone metastases is well tolerated and effective in decreasing bone pain, thus maintaining considerably high quality of life. PMID- 14750421 TI - [The effectiveness of ergotamine treatment in chronic headache disorders: a double-blind, randomized, crossover, placebo controlled trial]. AB - BACKGROUND: Postulated mechanism of migraine headache is a result of vascular spasm (primary) and dilatation with edema of the perivascular space (secondary). Thus we use drugs not the only during acute phase of headache but in prevention, also. Drugs containing ergotamine are well known and were used for years in the treatment of migraines. In this study we tried to investigate the role of multicomponent drugs containing ergotamine in the treatment of migraine in the adult workers group. MATERIALS AND METHODS: Experiments were performed with the use of double blind, randomized, crossover, placebo controlled study. Drug, Hydacorn, is a multicomponent drug where one tablet contains 1 mg of dihydroergo tamine, 40 mg of caffeine and 100 mg of diphenylhydantoine. 36 adults were enrolled (27 females and 9 males). Patients were examined 5 times every 2 weeks. During the 3rd visit the drug was changed to Hydacorn and placebo. RESULTS: During the 1st visit the intensity of pain did not differ significantly. After start of treatment the pain intensity did not differ significantly versus the 1st visit in placebo group. In the Hydacorn group we observed the significant reduction of pain intensity vs 1st visit and vs placebo. After drug change the significant reduction was present vs placebo (during the 5th visit only) and vs the 1st visit. Pain intensity during the 4th visit did not differ significantly vs placebo probably because of long action time for ergotamine. CONCLUSIONS: Our results suggest that Hydacorn is very effective and safe in the prevention of migraines. PMID- 14750422 TI - [The influence of birch pollen on adenoid hypertrophy in children with seasonal allergic rhinitis]. AB - The adenoid is a part of Waldeyer's ring, the basic function of which is antibody formation, they react later against a great variety of antigens. Pediatric adenoidal obstruction of the nasal airway is associated with significant morbidity and is a frequent indication for surgery, but its relationship to allergy has not been often studied. To examine the influence of atopy on the adenoid hypertrophy we studied 9 children ages 4 to 8 years, who had hypersensitivity to birch pollen and the seasonal allergic rhinitis. We conclude that allergy and sensitivity to birch pollen and probably to other kinds of alrallergens is an important risk factor for a AH in children. Therefore, early prevention of exposure to them may help reduce occurrence of AH. PMID- 14750423 TI - [Laboratory tests for HIV infection]. AB - This report describes commercially available tests for HIV diagnosis. It evaluates the sensitivity and specificity of tests and provides guidance for interpreting the test results. How use of them in HIV diagnosis is dependent on which kind of test was used, when the proper technique can to be used and how to interpret the results. The following diagnostic techniques are described: antibody related (EIA and Western blot), genetic test (PCR), viral protein estimation (p24 Ag), cell and viral culture. PMID- 14750424 TI - [The role of platelet glycoprotein receptors and their ligands: fibrinogen and von Willebrand factor in arterial thrombosis]. AB - The arterial platelet thrombus contribute to development of thrombotic complications of atherosclerosis: acute coronary syndrome, thrombotic strokes and exacerbations of peripheral arterial occlusive disease. Platelet adhesion and aggregation is mediated by interaction between platelet glycoprotein receptors GPI b/IX GPII b/III a and their ligands-adhesive proteins: von Willebrand factor (vWF) and/or fibrinogen. Pharmacological blockade of interaction between glycoproteins receptor and their ligands may offer an effective prevention of thrombotic complications. PMID- 14750425 TI - [Traumatic epidural hematoma in a patient with von Willebrand's disease]. AB - A case of traumatic epidural haematoma associated with von Willebrand disease (type 3) is reported. A 32-year-old man fell and hit his frontal region against the floor. CT scan on admission showed acute epidural haematoma of the right frontal region 45 mm in width and 60 ml volume. Considering moderate clinical symptomatology, a possibility of complication in connection with a congenital coagulation disorder and, above all, lack of patient's consent, prophylactic treatment with clotting factor concentrates was undertaken. Complete resorption of epidural haematoma has been obtained. In our opinion, in the presence of relative contraindication to operative intervention, even in cases with a large intracranial haematoma and inborn clotting deficiency prophylactic treatment may be considered. The following conditions must be met before such treatment is undertaken: prompt use of appropriate clotting factors, CT scan, clinical observation as well as possibility of prompt operative intervention in the neurosurgical department must all be available. PMID- 14750426 TI - [Unilateral exophthalmos during the course of cavernous sinus thrombosis: a case report]. AB - Ophthalmopathy, particularly its infiltrative form, is considered a characteristic feature of Graves' disease. However many other also life-treating conditions may be manifested as unilateral exophthalmos and lead to diagnostic problems. A case of a sixty six-year old patient with asymmetric exophthalmos and rapidly decreasing vision acuity is reported. Thyroid eye disease was suspected due to clinical symptoms (diplopy, impaired right eye motion) and thickening of extraocular muscles in ultrasound imaging. Further follow-up and orbits as well as head MRI imaging led to the final diagnosis of cavernous sinus thrombosis, complication of acute maxillary sinusitis. PMID- 14750427 TI - Arabis gemmifera is a hyperaccumulator of Cd and Zn. AB - Hyperaccumulators are essential for phytoremediation of heavy metals. In Europe and North America, many studies have been conducted to find more effective plants for phytoremediation of various pollutants. In Japan, this field of research has just recently come more into focus. A type of fern in Japan, Athyrium yokoscense, is well known as a hyperaccumulator of Cd and Zn. However, it is not suitable for phytoremediation because it is a summer green and grows slowly. Therefore, in order to find hyperaccumulators other than from A. yokoscense, we surveyed plants growing at polluted sites in Japan. We found that the Brassicae Arabis gemmifera is a hyperaccumulator of Cd and Zn, with phytoextraction capacities almost equal to Thlaspi caerulescens. PMID- 14750428 TI - Sulphur phytoaccumulation in plant species characteristic of Gypsiferous soils. AB - The SO4= toxicity in gypsiferous soils, which represent more than 100 million hectares worldwide, constitutes one of the major problems limiting world agricultural output. Currently, phytoremediation of SO4= is regarded, from agricultural and environmental standpoints, as one of the most effective alternatives for the decontamination and recovery of these soils. In this study, we analyzed the behavior of five plant species characteristic of gypsiferous soils (Gypsophila struthium, Helianthemun alypoides, H. squamatum, H. syriacum, and Lepidium subulatum) in order to establish the variability of these plants in terms of S-extraction mechanisms. Our results indicate that the species best adapted and with the densest population was Lepidium subulatum. This plant showed the highest concentration of total S and the greatest organic S, as well as the highest levels of amino acids and proteins. In this study, we propose that the incorporation of S into organic compounds is the essential metabolic process determining the tolerance of Lepidium subulatum to gypsiferous soils. In conclusion, the foliar accumulation of organic S could be defined a priori as one of the key characteristics in developing plants with a potential for regenerating zones with high SO4= concentrations. PMID- 14750429 TI - Aquatic and terrestrial plant species with potential to remove heavy metals from storm-water. AB - Remediation of storm-water polluted with heavy metals should be possible in percolation systems, ponds, or wetlands. The aim of this work was to find plant species for such systems that are efficient in the uptake of Zn, Cu, Cd, and Pb. Plants were collected from percolation and wetland areas and analyzed for heavy metal concentrations. Results showed that submersed and free-floating plants had the capacity to take up high levels of Cu, Zn, and Pb into their shoots. With roots having a concentration factor above 1, the terrestrial plants show efficient stabilization of Cd and Zn and emergent plants show corresponding stabilisation of Zn. In addition, Potamogeton natans, Alisma plantago-aquatica, and Filipendula ulmaria were used in a controlled experiment. The shoots of P. natans and the roots of A. plantago-aquatica were found to accumulate even higher concentrations of Zn, Cu, and Pb than found in the field-harvested plants. Similar results were found for Cd in shoots and Pb in roots of F. ulmaria. Our conclusion is that submersed plant species seem to be the most efficient for removal of heavy metals from storm-water. PMID- 14750430 TI - Degradation of crude oil in the rhizosphere of Sorghum bicolor. AB - Dissipation of petroleum contaminants in the rhizosphere is likely the result of enhanced microbial degradation. Plant roots may encourage rhizosphere microbial activity through exudation of nutrients and by providing channels for increased water flow and gas diffusion. Phytoremediation of crude oil in soil was examined in this study using carefully selected plant species monitored over specific plant growth stages. Four sorghum (Sorghum bicolor L.) genotypes with differing root characteristics and levels of exudation were established in a sandy loam soil contaminated with 2700 mg crude oil/kg soil. Soils were sampled at three stages of plant growth: five leaf, flowering, and maturity. All vegetated treatments were associated with higher remediation efficiency, resulting in significantly lower total petroleum hydrocarbon concentrations than unvegetated controls. A relationship between root exudation and bioremediation efficiency was not apparent for these genotypes, although the presence of all sorghum genotypes resulted in significant removal of crude oil from the impacted soil. PMID- 14750431 TI - Nickel and cobalt phytoextraction by the hyperaccumulator Berkheya coddii: implications for polymetallic phytomining and phytoremediation. AB - We investigated the potential of the South African high-biomass Ni hyperaccumulator Berkheya coddii to phytoextract Co and/or Ni from artificial metalliferous media. Plant accumulation of both metals from single-element substrates indicate that the plant/media metal concentration quotient (bioaccumulation coefficient) increases as total metal concentrations increase. Cobalt was readily taken up by B. coddii with and without the presence of Ni. Nickel uptake was, however, inhibited by the presence of an equal concentration of Co. Bioaccumulation coefficients of Ni and Co for the single element substrates (total metal concentration of 1000 micrograms g-1) were 100 and 50, respectively. Cobalt phytotoxicity was observed above a total Co concentration in plant growth media of 20 micrograms g-1. Elevated Co concentrations significantly decreased the biomass production of B. coddii without affecting the bioaccumulation coefficients. The mixed Ni-Co substrate produced bioaccumulation coefficients of 22 for both Ni and Co. Cobalt phytotoxicity in mixed Ni-Co substrate occurred above a total Co concentration of 15 micrograms g-1. When grown in the presence of both Ni and Co, the bioaccumulation coefficients of each metal were reduced, as compared to single-element substrate. This may indicate competition for binding sites in the root zone. The interference relationship between Ni and Co uptake demonstrated by B. coddii suggests a significant limitation to phytoextraction where both metals are present. PMID- 14750432 TI - Phytotransformation of benzotriazoles. AB - Plant roots interact with organic pollutants and some of these contaminants can be phytotransformed. Root uptake of 1-H-benzotriazole and its derivatives, tolyltriazole, 5-methyl benzotriazole, and 1-hydroxy benzotriazole was studied. At levels below the toxic threshold of about 100 mg/L, triazoles appear to be incorporated into plant tissue. Their concentration in the aqueous phase of the culture decreases with time and they cannot be extracted from the plant material using methanol. Hydroponic studies with sunflowers (Helianthus annuus) were used to investigate the behavior of the solution concentration versus time and to determine kinetic parameters for plant uptake of triazoles. Plants actively take up the triazoles at a rate greater than predicted by transpiration stream concentration factor and plant-water uptake. Analyses of the data for phytotransformation rate versus concentration were performed to establish the kinetic model for the removal process. Except for 1-hydroxy-benzotriazole, triazole disappearance in plant systems followed the Michaelis-Menten kinetic model (commonly found for enzyme-catalyzed reactions) better than a first order rate model. However, the fit for the first order model was improved when normalizing to the plant fresh weight, which was assumed to be an approximate measure of the changing root surface area. Experiments with other plant species are in progress. PMID- 14750433 TI - Purification characteristics of golden pothos for atmospheric gasoline. AB - Previous studies have shown that plants have the ability to purify various atmospheric chemicals. Gasoline is one of the more serious pollutants. Soil and atmospheric pollution caused by gasoline is increasing due to the widespread use of automobiles. In this article, the purification characteristic of the pothos plant for atmospheric gasoline is investigated using a tin oxide gas sensor. The purification rate (Pr), defined as the purification ability per hour as described by a differential coefficient, has a maximum value at longer time intervals as the pollutant concentration becomes higher. Pr can be represented by an exponential function of lapsed time and its characteristic in soil is similar. A golden pothos plant growing in a 30-cm diameter pot of was placed in a 300-I experimental chamber to examine its purification ability. Pr had a maximum value 40 h after a 0.04-ml injection of gasoline into the chamber. The total purification ability (Pa) is also used in this study and is derived using the peak value (h) and the full width (tw) at half maximum of the tin oxide gas sensor characteristic, namely Pa = h/tw x 100. The Pa of the pothos for gasoline was about 7, with the value decreasing as the pollutant concentration increased. PMID- 14750434 TI - The molecular basis of infectious diseases: pathogenicity islands and other mobile genetic elements. A review. AB - Bacterial genomes generally consist of stable regions termed core genome, and variable regions that form the so-called flexible gene pool. The flexible part is composed of bacteriophages, plasmids, transposons as well as unstable large regions that have been termed genomic islands. Genomic islands encoding virulence factors of pathogenic bacteria have been designated "pathogenicity islands". Pathogenicity islands were first discovered in uropathogenic Escherichia coli and presently more than 30 bacterial species carrying pathogenicity islands have been described. This review summarises the current knowledge on bacterial genomic islands and their general features, and discusses their putative role in the evolution of microbes in the light of genomics of pathogenic bacteria. PMID- 14750435 TI - Increased salt- and nisin-sensitivity of pressure-injured bioluminescent Listeria monocytogenes. AB - Suspensions of a bioluminescent (luxAB) transformant of Listeria monocytogenes in pH 7.0 phosphate buffer were pressurised and the effect of the pressure treatment was monitored by plate counting. When the bacteria were suspended in NaCl- and nisin-free buffer the number of colony forming units (CFU) decreased by 3 and 6 log cycles after 300 MPA for 10 and 30 min, respectively. Supplementing the plating medium with 5% NaCl did not influence the colony forming capacity of non pressurised cells, however, CFU of residual populations after respective treatments of 300 MPa for 10 and 30 min were reduced by a further 2 and 3.5 log cycles in case of salt containing plates. Nisin-addition to the plating medium caused less than one log unit decrease in the CFU of the non-pressurised population. However, the CFU of 10 min-pressurised sample was 4 log cycles less in the nisin-containing plates than in the nisin-free ones, whereas no colonies were formed in the nisin-containing plates even when 1 ml was inoculated from the originally 10(10) CFU/ml population after 300 MPa for 30 min. The luciferase activities (bioluminescence intensities) decreased concomitant with the reduction of the viable cell counts, however, they were approx. 0.6-0.8 log units less in the presence of 5% NaCl in the pressurised suspension than those expected from the previously established linear correlation between the logarithmic light outputs and the logarithmic viable cell counts. PMID- 14750436 TI - Increased levels of tumor necrosis alpha and soluble vascular endothelial adhesion molecule-1 in the cerebrospinal fluid of patients with connective tissue diseases and multiple sclerosis. AB - The aim of the present study was to investigate the serum and cerebrospinal fluid (CSF) concentrations of tumor necrosis factor alpha (TNF-alpha) and soluble vascular cell adhesion molecule-1 (sVCAM-1) in patients with primary progressive form of multiple sclerosis (MS) and in patients with connective tissue diseases (CTDs) complicated with central nervous system (CNS) involvement. Stimulation of sVCAM-1 release by TNF-alpha was demonstrated on endothelial cells of brain vessels. We intended to present the TNF-alpha stimulated elevation of sVCAM-1 in the serum and CSF in any cases of CNS lesion. Fifty patients with several CTDs complicated with neuropsychiatric symptoms and 25 MS patients with primary chronic progressive form of the disease were selected. Determinations of TNF alpha and sVCAM-1 were performed using ELISA methods. TNF-alpha and sVCAM-1 concentrations were elevated in the CSF of all patients, intrathecal synthesis of sVCAM-1 was demonstrated in MS patients. The changes in the TNF-alpha and sVCAM-1 concentrations were independent from the clinical manifestations, immunoserological changes and quality of neuropsychiatric symptoms of the CTDs. The stimulatory effect of TNF-alpha was more pronounced in the CSF of MS patients. PMID- 14750437 TI - Comparison of traditional and molecular typing methods of Escherichia coli O157. AB - An account is given using typing methods and detection of virulence genes of different serotypes of Escherichia coli isolated in Hungary. By hybridization using SLT-I and SLT-II probes and PCR method using stx1-2, eae and ehx primers we could differentiate O157 strains of different serotypes into eight (stx, eae, ehxA positive; stx, eae positive; stx, ehxA positive; stx positive; eae, ehxA positive; eae positive; ehxA positive; stx, eae, ehxA negative) types. The discriminatory power of phage typing proves to be much higher than that of the plasmid profile. RAPD typing with different primers could confirm or exclude the subtypes identity of the isolated E. coli O157 serotypes. Escherichia coli O157:HNM isolates could be sorted in six different phage types and six different RAPD types with ERIC-1, in five RAPD types with ERIC-2 and in seven types with M13 primers. Escherichia coli O157:H7 showed six different phage types and three RAPD types with ERIC-1 and ERIC-2 and five types with M13 primers. According to our results the standard PFGE protocol [32] gives the opportunity to differentiate epidemiologically independent but evolutionary related or unrelated isolates, but the practical value of PFGE method for epidemiological purposes must be confirmed by other or more restriction enzymes or using an other protocol. Summarizing our results we suggest the use of phage and RAPD typing and in doubtful cases the PFGE method. PMID- 14750438 TI - The use of spiral loops in serological and virological micro-methods. 1955. PMID- 14750439 TI - QSAR analysis of antibacterial activity of some 4-aminodiphenylsulfone derivatives. AB - QSAR studies on a set of 36 congeners of 4-aminodiphenylsulfone derivatives with measured inhibition potencies of dihydropterate synthase were made using multiple regression analysis. Conformational entropy in combination with indicator parameters gave excellent results. PMID- 14750441 TI - Molecular mechanisms of virus spread and virion components as tools of virulence. A review. AB - Despite of differences in replication strategy among virus families, some basic principles have remained similar. Analogous mechanisms govern virus entry into cells and the use of enzymes which direct the replication of the virus genome. The function of many cell surface receptors (such as glycosoaminoglycans, glycoproteins, proteins) which interact with viral capsid proteins or envelope glycoproteins has recently been elucidated. The list of cellular receptors (Table I) is still far from being final. The capsid components, similarly as the envelope glycoproteins, may form specific pocket like sites, which interact with the cell surface receptors. Neutralizing antibodies usually react with antigenic domains adjacent to the receptor binding site(s) and hamper the close contact inevitable for virion attachment. In the case of more complex viruses, such as herpes simplex virus, different viral glycoproteins interact with several cellular receptors. At progressed phase of adsorption the virions are engulfed into endocytic vesicles and the virion fusion domain(s) become(s) activated. The outer capsid components of reoviruses which participate in adsorption and fusion may get activated already in the lumen of digestive tract, i.e. before their engulfment by resorptive epithelium cells. Activation of the hydrophobic fusion domain(s) is a further important step allowing to pass through the lipid bilayer when penetrating the cell membrane in order to reach the cytosol. Activation of the virion fusion domain is accomplished by a conformation change, which occurs at acid pH (influenza virus hemagglutinin, sigma 1 protein of the reovirus particle) and/or after protease treatment. The herpes simplex virus fusion factors (gD and gH) undergo conformation changes by a pH-independent mechanism triggered due to interaction with the cell surface receptor(s) and mediated by mutual interactions with the viral envelope glycoproteins. The virion capsid or envelope components participating in the entry and membrane fusion are not the only tools of virulence. The correct function of virus coded proteins, which participate in replication of the viral genome, and/or in the supply of necessary nucleotides, may be very essential. In the case of enteroviruses, which RNA interacts with ribosomes directly, the correct configuration of the non-coding viral RNA sequence is crucial for initiation of translation occurring in the absence of the classical "cap" structure. PMID- 14750440 TI - Molecular microbiology of gut bacteria: genetic diversity and community structure analysis. AB - Recently developed molecular biology approaches make possible the detailed genetic, taxonomic and ecological examination of microorganisms from various habitats. Animal gut represents one of the most complex microbial ecosystems with a large degree of microbial biodiversity present. Bacteria inhabiting the gut usually play important roles in metabolic transformations of substrates and sometimes, e.g. in ruminants, they make the basis for an obligate symbiosis with the host. Here we discuss molecular microbiology as a strategy for examination of gut bacteria, concentrating on a typical and in such environment dominant group of strictly anaerobic Gram-negative bacteria from the phylogenetic group Cytophaga/Flexibacter/Bacteroides. The bacteria from the genus Prevotella are the most abundant Gram-negative bacteria in the rumen and form a distinctive phylogenetic cluster, clearly separated from prevotellas isolated from other ecological niches. They may represent a good choice for a model organism in genetic manipulation experiments and for studies of gene transfer mechanisms taking place in the gut. The molecular tools for detection and monitoring of ruminal prevotellas are discussed. PMID- 14750442 TI - The ion channels coded by viruses. AB - Pathogenicity and virulence are multifactorial traits, depending on interaction of viruses with susceptible cells and organisms. The ion channels coded by viruses, viroporins, represent only one factor taking part in the cascade of interactions between virus and cell, leading to the entry of virus, replication and to profound changes in membrane permeability. The M2 protein from influenza A virus forms proton-selective, pH-regulated channel involved in regulating vesicular pH, a function important for the correct maturation of HA glycoprotein. The NB glycoprotein of influenza B viruses is an integral membrane protein with an ion channel activity. The CM2 protein of influenza C virus is an integral membrane glycoprotein structurally analogous to influenza A virus M2 and influenza B virus NB proteins. The picornavirus 3A protein is involved in cell lysis and shows homology with other lytic proteins. Vpu is an oligomeric integral membrane protein encoded by HIV-1, which forms ion channels. The togavirus 6K protein shows structural similarities with other viroporins. PMID- 14750443 TI - Virus and host factors that mediate the clinical and behavioral signs of experimental herpetic encephalitis. A short auto-review. AB - Experimental models that mimic the clinical syndrome of human viral encephalitis and represent HSV-1 neurotropism were utilized to investigate neuro-pharmacologic changes mediating clinical and behavioral manifestations of encephalitic infection of the central nervous system with HSV-1-induced rapid activation of the hypothalamic--pituitary--adrenocortical (HPA) axis and production of brain derived interleukin-1 (IL-1) and prostaglandin E2 (PG-E2), independently of viral replication. HSV-1 infection induced clinical signs of fever, motor hyperactivity and aggressive behavior. These manifestations were dependent on a permissive action of circulating glucocorticoids and not related to the degree of viral replication in the brain. Hyperthermia and HPA axis activation were also specifically dependent on HSV-1-induced brain IL-1 and PG-E2. The chronic neurological sequel or fatal outcome of HSV-1 encephalitis may be due to viral replication and brain tissue destruction, which are dependent on virus encoded virulence genes. In contrast, the clinical and behavioral signs in the acute phase are a result of activation of neurochemical systems, including cytokines, prostaglandinds and catecholamines. Circulating glucocorticoids play an essential role in mediating the physiologic actions of HSV-1-induced brain products and the clinical syndrome of encephalitis. PMID- 14750444 TI - Vancomycin-resistant Enterococcus faecalis colonization during recovery from Neisseria meningitidis cerebrospinal meningitis. Case report. AB - A 19-year-old man had been admitted to the Hospital because of septic shock and large scale suffusions all over the body. The pathogen had proved to be Neisseria meningitidis serogroup C. In his stabilization period two superinfectious attacks arose. One of them was a bacteremia, caused by a vancomycin-sensitive Enterococcus faecium. The second was a wound infection in his deep colliquating necrotised tissue of the heel. Vancomycin-resistant Enterococcus faecalis (VREF) was isolated from this lesion with some Gram-negative opportunistic pathogens. The strain contained the vanA gene. After systemic and topical treatment, furthermore plastic surgical interventions the patient recovered. This is the second report on VREF from Hungary colonizing/infecting a patient with an underlying disease. PMID- 14750445 TI - [The human measure]. AB - In 1800 the new French republic introduced the metric system to replace a multitude of different measures. Some old units for length, such as the foot, were based on the human body. In medicine, the severity of a disease can be measured at several different levels; (a) the biological process; (b) the function of body parts; (c) the individual's activities; and (d) social participation. In the current quest for objectivity, physicians sometimes prefer measurements that are closely related to physical science, whereas patients are more concerned about the restriction of their activities. There are different scales for measuring the range of human activities, for example, according to speciality or even disease. There is a need for a generic measure of health in relation to activities and this should preferably be calibrated on a linear scale. PMID- 14750446 TI - [Patient information after a stroke: the needs in relation to the different phases]. AB - A 78-year-old woman felt uncertain 4 days after a stroke, while a 65-year-old man wanted to do too much 6 weeks after a stroke. An 81-year-old man was depressive 6 months after a stroke. By paying attention to the need for information and the burden on the carer, the general practitioner tried to improve the quality of life in both cases. The patient's and carer's ability to cope with post-stroke disability is enhanced by providing adequate information. The three consecutive phases in the post-stroke process have specific characteristics and corresponding needs for information: clinical information in the acute phase, coping information in the rehabilitation phase, and practical information in the chronic phase. In each phase, the information should be tuned to the individual needs of the patient and the carer. Coordination with other professionals is necessary. PMID- 14750447 TI - [Decrease in hospital admissions due to cardiovascular diseases]. AB - Since the late 1970s there has been a decline in cardiovascular mortality in the Netherlands. In recent years, the number of hospital admissions due to cardiovascular disease has decreased, whereas short-stay admissions and outpatients' visits are increasing. One explanation for this phenomenon, suggested by Koek et al. in this issue, is the limited availability and closure of hospital beds in recent years. However, there are other possible reasons for the decrease in cardiovascular mortality. First of all, there has been a substantial improvement in survival following acute myocardial infarction, mainly as a result of advances in the treatment. As all patients with an acute myocardial infarction are admitted anyway, the observed decrease in hospital admissions due to acute myocardial infarction reflects a decrease in the incidence as opposed to a decrease in the availability of hospital beds. Secondly, primary preventive measures aimed at modifying risk factors have been shown to result in a clear decrease in cardiovascular mortality. Lastly, mortality and morbidity due to chronic cardiovascular disease, such as heart failure, have decreased significantly. The range of options to adequately treat this category of patients has expanded considerably with, for example, the introduction of new drugs and biventricular pacemakers. There are many factors, other than the closure of hospital beds, which could explain the observed decline in the number of admissions due to cardiovascular disease. PMID- 14750448 TI - [High dose chemotherapy and stem cell transplant in patients with operable breast cancer and a poor prognosis; not indicated for the time being]. AB - Early breast cancer with multiple tumour-positive axillary lymph glands is still characterised by a considerable mortality despite improvements in surgery, radiotherapy, chemotherapy and endocrine treatment. High-dose chemotherapy and bone-marrow support instead of the usual chemotherapy seemed to be a very promising approach back in the 1980s. Several phase-II studies supported this view and therefore phase-III studies were carried out. In a Cochrane review published at the start of 2003, a meta-analysis of several phase-III studies revealed that the results after high-dose chemotherapy were not significantly better than those after the usual treatment. Two large multicentre studies, one performed in the Netherlands and one in the USA, have recently been published. Neither of the two studies revealed survival differences, although one study demonstrated a significant benefit in actuarial 5-year relapse-free survival. Therefore, the routine clinical application of high-dose chemotherapy cannot be advocated yet. PMID- 14750449 TI - [Nutrition and health--safety of new types of food]. AB - In Europe it is currently a statutory requirement that all novel foods must first be tested before they can be put on the market to establish that they pose no hazard to consumers. A novel food is a food or food ingredient which was not used to a significant degree on the European market prior to 1997 and which falls within one of the categories described in a specific European regulation. The procedure used to establish the safety of such a novel food involves a request for authorization. The company compiles a safety dossier containing a comprehensive report on the characteristics of the substance, details of toxicological studies and any other relevant information. The company submits the dossier to the relevant authority in one of the European member states. In the Netherlands, this is the Ministry of Health, Welfare and Sport. The Ministry then arranges for an initial assessment to be carried out. In the Netherlands the Committee on Safety Assessment of Novel Foods (Dutch acronym: VNV), part of the Health Council, conducts the assessment. Following the completion of the assessment, other member states may carry out their own assessments. Finally, the member states make a collective decision on whether or not to authorise the marketing of the product. The VNV assessment takes a case-by-case and step-by step approach. Each request for authorization is individually examined to determine the types of research data that will be needed to reach a verdict. A short-track procedure or 'notification' is available for products that are substantially equivalent to existing foods. The VNV Committee operates as transparently as possible. All dossiers are available for inspection. The Committee's advisory reports are available to the public and committee members provide details of their personal interests. PMID- 14750450 TI - [Further development of 'Enuresis nocturna' guidelines for children with persistent problems]. AB - To achieve dryness in children who suffer from persistent bedwetting, it is important to find out which factors play a role in their bedwetting and why previous treatment has failed. The use of a micturition diary is essential. The child's problems and needs have to be identified and treated individually. The enuresis alarm, with the proper guidance, is the preferred form of initial treatment. Desmopressin is particularly suitable in cases of nocturnal polyuria or if the use of the alarm is unfeasible. If the alarm does not have any effect within two weeks or if a rapid result is important, a combination of desmopressin and the alarm is advisable. Bedwetting combined with daytime micturition problems is often indicative of a small bladder capacity and/or detrusor instability. In these children bladder training and/or treatment with an anticholinergic drug (possibly combined with desmopressin) may be effective. PMID- 14750451 TI - [Diagnostic image (171). A man with a painful red ear. Relapsing polychondritis]. AB - A 50-year-old man presented with a painful and red swollen right ear, fever, iridocyclitis and a peripheral facial nerve palsy, due to relapsing polychondritis. Immunosuppressive therapy was successful. PMID- 14750452 TI - [Trends in cardiovascular morbidity and mortality in the Netherlands, 1980-2000]. AB - OBJECTIVE: To describe and interpret trends in cardiovascular morbidity and mortality among the Dutch population from 1980 to 2000, in particular the decline in hospital admissions for cardiovascular disease during the second half of the 1990s. DESIGN: Descriptive. METHOD: An analysis of trends in hospital admissions and cardiovascular mortality for the period 1980 to 2000 was performed using Dutch hospital admissions and mortality data, specified according to age and sex. These data were taken from the National Medical Register of hospital admissions maintained by Prismant and from the national statistics on the causes of mortality maintained by Statistics Netherlands. RESULTS: Since the 1970s, the mortality due to cardiovascular diseases has gradually declined. The age-adjusted mortality has declined by 36% for men and 33% for women. The decline is evident for both acute and chronic diseases. After an initial increase, hospital admission rates for cardiovascular disease showed a marked decline during the second half of the 1990s. The age-adjusted hospital admission rate has declined by 15% for men and 13% for women. This decline was seen in all age groups and for both acute and chronic diseases. CONCLUSION: The recent decrease in hospital admissions for cardiovascular disease during the second half of the 1990s appears mainly to be the consequence of a shift from hospital admissions to day care and outpatients' clinics. PMID- 14750453 TI - [Empty sella syndrome as the cause of panhypopituitarism]. AB - In two patients, men aged 78 and 42 years respectively, an empty-sella syndrome was found. The first patient presented with chronic fatigue, in the second the empty sella was the late result of prior neurosurgery followed by external irradiation. Both suffered from panhypopituitarism. In both cases the diagnosis was confirmed following laboratory tests and MRI. Hormone-replacement therapy was found to provide adequate treatment. The course of the empty-sella syndrome is usually benign and with adequate hormone-replacement therapy a good quality of life is maintained. PMID- 14750454 TI - [A patient with an extremely severe immunohaemolytic anemia due to diclofenac]. AB - A 77-year-old male was diagnosed with immunohaemolytic anaemia. He presented with fever, jaundice and oliguric renal insufficiency. He had been taking diclofenac for three days. Additional examinations and tests revealed a Coombs-positive haemolytic anaemia and thrombopenia. The micro-card gel-agglutination test demonstrated antibodies against diclofenac metabolites. After discontinuation of diclofenac and short-term treatment with prednisone and haemodialysis for two days, the patient made an almost full recovery. Haemolytic anaemia caused by diclofenac is rare. It can be caused by autoantibodies against erythrocytes and complement-fixing antibodies that bind to erythrocytes only in the presence of diclofenac or its metabolites. A timely diagnosis-supported by the appropriate laboratory tests--is of critical importance in this rare but extremely serious complication of the use of diclofenac. The treatment consists of withdrawal of diclofenac and supportive measures, possibly in combination with immunosuppressants. PMID- 14750455 TI - [The guideline 'Detection of visual disorders at the age of 0-19' from the Dutch Association of Child Health Care]. PMID- 14750456 TI - [Euthanasia and other medical decisions at the end of life in the Netherlands in 1990, 1995 and 2001]. PMID- 14750457 TI - [Reports of transfusion incidents: experiences from the first year of hemovigilance in the region of the former ZWN (South West Netherlands) blood bank in Rotterdam]. PMID- 14750458 TI - Pressuring insurers to pay for medically unnecessary treatments. PMID- 14750459 TI - Time to antidepressant discontinuation: a comparison of controlled-release paroxetine and immediate-release selective serotonin-reuptake inhibitors. AB - Despite advances in depression therapy, early treatment discontinuation with antidepressants remains high and contributes to poor clinical outcomes. In clinical trials, treatment discontinuation with controlled-release paroxetine was similar to placebo and significantly better than immediate-release paroxetine. To examine whether this benefit noted in clinical trials translated into longer therapy duration in practice settings, survival-analysis models were constructed to compare the time to treatment discontinuation of controlled-release paroxetine with that of immediate-release selective serotonin-reuptake inhibitors (SSRIs). Survival analysis indicated that patients receiving controlled-release paroxetine were 28% less likely to discontinue therapy during a 180-day period when compared with patients receiving immediate-release SSRIs. PMID- 14750460 TI - A decade of evolution in disease management. PMID- 14750461 TI - Evaluating the role of confounding in pharmacoeconomic studies. AB - Confounding may be present in nonrandomized etiological research involving human populations. It can result in erroneous conclusions about the effect of exposure on a disease outcome or about any form of causality between predictors and outcomes. Confounding can wholly or partially account for the apparent effect of the risk factor under consideration or mask the underlying, true association. Not controlling for the effects of confounding can lead to biased results, thus compromising the validity of study conclusions. The three goals of this article are: (1) to define a confounder or a confounding variable, (2) to discuss strategies for controlling the effects of confounding, and (3) to illustrate the perverse effects of confounding with the help of an example. PMID- 14750462 TI - Disease management and antikickback regulations. PMID- 14750464 TI - Beyond doer and done to: an intersubjective view of thirdness. AB - Analytic work based on the intersubjective view of two participating subjectivities requires discipline rooted in an orientation to the structural conditions of thirdness. The author proposes a theory that includes an early form of thirdness involving union experiences and accommodation, called the one in the third, as well as later moral and symbolic forms of thirdness that introduce differentiation, the third in the one. Clinically, the concept of a co-created or shared intersubjective thirdness helps to elucidate the breakdown into the twoness of complementarity in impasses and enactments and suggests how recognition is restored through surrender. PMID- 14750465 TI - Subjectivity, objectivity, and triangular space. AB - The author reviews his ideas on subjectivity, objectivity, and the third position in the psychoanalytic encounter, particularly in clinical work with borderline and narcissistic patients. Using the theories of Melanie Klein and Wilfred Bion as a basis, the author describes his concept of triangular space. A case presentation of a particular type of narcissistic patient illustrates the principles discussed. PMID- 14750466 TI - The relational unconscious: a core element of intersubjectivity, thirdness, and clinical process. AB - The relational unconscious is the fundamental structuring property of each interpersonal relation; it permits, as well as constrains, modes of engagement specific to that dyad and influences individual subjective experience within the dyad. Three usages of the concept of thirdness are delineated and contrasted with the concept of the relational unconscious, which, it is suggested, has the advantage of being both consistent with existing views of unconscious processes and more directly applicable to therapeutic concerns. Enactments and intersubjective resistances are viewed as clinical manifestations of the relational unconscious, and the therapeutic action of psychoanalysis results, in part, from altering the structure of the relational unconscious that binds analysand and analyst. PMID- 14750467 TI - Thirdness and psychoanalytic concepts. AB - Many psychoanalytic concepts lend themselves to the notion of thirdness. Starting from a basis of Freudian thought, the author discusses some of the elaborative contributions of Winnicott, Lacan, and Bion, as well as the ideas of Saussure and Peirce, noting how all these incorporate an appreciation of the value and relevance of thirdness in both the theory and practice of psychoanalysis. PMID- 14750468 TI - Solving the problems of duality: the third and self-consciousness. AB - Locating the concept of the third in the debate about countertransference that began in the 1950s, the authors maintain that it originated to solve problems stemming from the recognition that the analytic encounter takes place between two individual subjects. This recognition can lead to discomfort for the analyst, once objective criteria to interpret reality have been lost due to adhesion to a dialectical constructionist perspective; it also implies a deeper involvement arising from the abandonment of neutrality. The concept of the third is often invoked to help avoid these risks. However, the authors maintain that only the human subject itself can grasp the self reflexively; this view has a referent in the Hegelian concept of self-consciousness and is also supported by the findings of infant research. PMID- 14750469 TI - The analytic third: implications for psychoanalytic theory and technique. AB - The author views the analytic enterprise as centrally involving an effort on the part of the analyst to track the dialectical movement of individual subjectivity (of analyst and analysand) and intersubjectivity (the jointly created unconscious life of the analytic pair--the analytic third). In Part I of this paper, the author discusses clinical material in which he relies heavily on his reverie experiences to recognize and verbally symbolize what is occurring in the analytic relationship at an unconscious level. In Part II, the author conceives of projective identification as a form of the analytic third in which the individual subjectivities of analyst and analysand are subjugated to a co-created third subject of analysis. Successful analytic work involves a superseding of the subjugating third by means of mutual recognition of analyst and analysand as separate subjects and a reap-propriation of their (transformed) individual subjectivities. PMID- 14750470 TI - The third in mind. AB - Various aspects of the occult as they relate to psychoanalysis are discussed in this article. Drawing on both Freud's writings and Granoff and Rey's (1983) work on the occult in Freudian thought, the author considers the concept of co thinking and its manifestations in clinical work. The psychoanalytic third is viewed in the context of the occult element known as thought transference, or thought transmission, and is also considered as it bears on psychoanalytic supervision. PMID- 14750471 TI - The third position: reflections about the internal analytic working process. AB - The development of a third position is a key aspect of the analyst's ability to survive in the analytic relationship. In exploring the value of the third position, the author discusses other models relevant to clinical work, including the analytic-therapeutic position, the internal analyst, the alive analytic contact, and the phobic position. A case example illustrates these models and forms the basis for further discussion of the internal analytic working process. PMID- 14750472 TI - The third: a brief historical analysis of an idea. PMID- 14750473 TI - Autogenous free tooth transplantation with a two-stage operation technique. AB - If tooth transplantation is to succeed, it is crucial to preserve the vitality of the cells on the root surface of the tooth transplant. Insufficient postoperative nutrition to the cells on the root surface of the tooth transplant was thought to contribute to their devitalization of these cells. Impaired nutrition may be a result of poor contact between the recipient bed and the root surface of the transplanted tooth, and development of an interposed blood clot. To improve postoperative nutrition to the root surface cells, teeth were transplanted to the recipient beds in which the tissues were regenerated during a 14 day period, i.e., using the two-stage transplantation technique. In a clinical study of this technique, a total of 95 autogenous teeth with fully developed roots were transplanted in 84 patients, and examined both clinically and radiographically for up to 13 years after the transplantation. In a dog model, a comparative experimental study was made between teeth transplanted to beds left to heal for 5 days and teeth transplanted to beds prepared immediately before the transplantation. The clinical study showed a low prevalence of tooth graft loss and root resorption even when infection of the root canal occurred. Periodontal attachment loss of less than 3 mm was found in 97% of 6 defined surfaces around the transplanted teeth. Transplanted teeth which were later extracted were often hypermobile, and signalled pain when provoked with heavy loading. Excessive extraction trauma, fixation failure, and excessive plaque accumulation after transplantation were all shown to be detrimental to tooth transplant. Transplanted teeth were used as abutments for fixed partial dentures and provided the necessary dental support for crowns and bridges even in patients with atrophic alveolar bone. The experimental histological study showed no differences between test and control teeth in terms of the prevalence of root resorption, which was suspected to be caused by traumatic injuries to the roots during extraction and non-rigid fixation of the transplanted teeth. PMID- 14750474 TI - The Malawi medical school--a success story. PMID- 14750475 TI - Dispensing with doctors? PMID- 14750476 TI - Ethics, litigation and teaching of anatomy. PMID- 14750477 TI - Mountain rescue--nerve blocks, helicopters, hypothermia and hypotension. PMID- 14750478 TI - Mountain rescue--nerve blocks, helicopters, hypothermia and hypotension. PMID- 14750479 TI - Legal bone throwing has doctors hopping mad. PMID- 14750480 TI - AIDS--why we must do more. PMID- 14750481 TI - Legislation to 'CON' doctors? PMID- 14750482 TI - Did we really help break the HIV prevalence wave? PMID- 14750483 TI - Results of WHO inquiry into SARS leave grey areas.. PMID- 14750484 TI - Developed world warned to avoid complacency over SARS. PMID- 14750485 TI - Landmark ruling by competition commission. PMID- 14750486 TI - WHO welcomes research showing effectiveness of AIDS treatment. PMID- 14750487 TI - Aspen to manufacture AIDS drugs for Clinton Foundation. PMID- 14750488 TI - Human resource requirements for introducing cervical screening--who do we need where? PMID- 14750489 TI - Antimycobacterial activity of indigenous South African plants. PMID- 14750490 TI - Whither the AIDS epidemic--or lies, damned lies and statistics? PMID- 14750491 TI - The living situations of orphans in periurban Blantyre, Malawi. PMID- 14750492 TI - Intracardiac air--the 'hospital killer' identified? Case reports and review of the literature. AB - Venous access is an essential part of caring for the sick neonate. The primary problem with catheters, whether peripherally or centrally placed, is the difficulty in maintaining them, the development of phlebitis and systemic infection, and fluid extravasation. A lesser known complication is the development of venous air embolism (VAE), as described in the 4 cases presented. We agree with others that VAE in newborn infants may occur more frequently than expected and emphasise the fact that it is preventable and that careful attention must be given to the techniques of preparing venous infusions. As health professionals (medical and nursing) we should take a harder line and regard these events as medically negligent until proven otherwise. We should take full responsibility for equipment, the connections, the infusate and the monitoring thereof. Unfortunately, the prognosis for this condition remains poor and it is unclear whether an increased awareness of this condition would influence outcome. Manufacturers of intravenous fluids should produce products devoid of air in order to reduce the risk of venous air embolism. PMID- 14750493 TI - Psychopathology and coping in recently diagnosed HIV/AIDS patients--the role of gender. AB - BACKGROUND: Although there is growing literature on the psychological responses to and the psychopathology associated with HIV/AIDS, few investigations have focused on the role of gender. This study compared psychiatric morbidity, coping responses, and disability in male and female outpatients recently diagnosed with HIV/AIDS. METHOD: One hundred and forty-nine patients (44 male, 105 female) with HIV/AIDS (mean +/- standard deviation (SD) months since diagnosis 5.8 +/- 4.1) attending an infectious diseases clinic at Tygerberg Hospital, Cape Town, were evaluated. Subjects were assessed using the MINI International Neuropsychiatric Interview (MINI), the Carver Brief COPE, and the Sheehan Disability Scale. In addition, negative life events and risk behaviours were evaluated. RESULTS: Fifty six per cent of patients were diagnosed with a psychiatric disorder, most commonly major depression (34.9%), dysthymic disorder (21.5%), post-traumatic stress disorder (14.8%), and alcohol dependence (10.1%). There were no significant gender differences in the prevalence of mood disorders in the sample. Men, however, were more likely than women to meet diagnostic criteria for alcohol abuse or dependence, and to engage in certain risky sexual behaviours. Women were more likely to suffer from post-traumatic stress disorder, and to use coping strategies of planning and religion to deal with the illness. There were no significant gender differences in disability. CONCLUSION: Psychiatric disorders are common in recently diagnosed HIV/AIDS patients in South Africa. Clinicians should be aware of the high prevalence of mood disorders in both men and women, and of gender-different responses such as increased alcohol and substance use and more risky sexual behaviour in men. PMID- 14750494 TI - Antinuclear antibody testing in obstetric patients. AB - OBJECTIVES: To assess possible associations between the presence of antinuclear antibodies (ANAs) and pregnancy outcome in order to determine the significance of this test in obstetric practice. METHODS: A case-control study was performed on 408 patients admitted to an obstetric high care unit and on whom ANA testing was consecutively performed. The study group consisted of 46 patients who tested positive for ANAs and a control group of 92 patients who tested negative for ANAs. In addition to demographic data, indications for admission and pregnancy outcome were compared between the two groups. RESULTS: Of the 46 patients with a positive ANA result, 28 had an antinuclear pattern, 13 an anticytoplasmic pattern and 5 an antinuclear and an anticytoplasmic pattern. No significant differences were observed between the two groups (ANA-positive and negative) with regard to demographic data, indication for admission, clinical and laboratory data, and pregnancy outcome. The patients were also tested for anticardiolipin antibodies, and significantly more patients with severe pre-eclampsia tested positive (24% versus 4.7%, p = 0.01). No difference in HIV status and presence of autoantibodies was found between the two groups. CONCLUSION: The presence of ANAs was not associated with adverse pregnancy outcome. Therefore routine patient testing for ANAs in an obstetric high-care unit is not recommended. PMID- 14750495 TI - The history and pathology of crucifixion. AB - In antiquity crucifixion was considered one of the most brutal and shameful modes of death. Probably originating with the Assyrians and Babylonians, it was used systematically by the Persians in the 6th century BC. Alexander the Great brought it from there to the eastern Mediterranean countries in the 4th century BC, and the Phoenicians introduced it to Rome in the 3rd century BC. It was virtually never used in pre-Hellenic Greece. The Romans perfected crucifion for 500 years until it was abolished by Constantine I in the 4th century AD. Crucifixion in Roman times was applied mostly to slaves, disgraced soldiers, Christians and foreigners--only very rarely to Roman citizens. Death, usually after 6 hours--4 days, was due to multifactorial pathology: after-effects of compulsory scourging and maiming, haemorrhage and dehydration causing hypovolaemic shock and pain, but the most important factor was progressive asphyxia caused by impairment of respiratory movement. Resultant anoxaemia exaggerated hypovolaemic shock. Death was probably commonly precipitated by cardiac arrest, caused by vasovagal reflexes, initiated inter alia by severe anoxaemia, severe pain, body blows and breaking of the large bones. The attending Roman guards could only leave the site after the victim had died, and were known to precipitate death by means of deliberate fracturing of the tibia and/or fibula, spear stab wounds into the heart, sharp blows to the front of the chest, or a smoking fire built at the foot of the cross to asphyxiate the victim. PMID- 14750496 TI - In vitro evaluation of the anti-prionic activity of newly synthesized congo red derivatives. AB - "Transmissible Spongiform Encephalopathies" (TSE) are a group of degenerative progressive fatal disorders of the CNS, affecting both humans and animals. The main pathogenic event is the conversion of cellular prion protein from the normal, enzyme-sensitive (PrPsen), to the insoluble proteinase K-resistant isoform (PrPres). Since the new juvenile variant of Creutzfeldt-Jakob disease (vCJD) is probably due to the transmission of Bovine Spongiform Encephalopathy (BSE) prion protein to man, therapeutic and preventive compounds for animals and humans are urgently needed. Congo Red (benzidine-diazo-bis-1-naphthylamine-4 sulfonic acid sodium salt, CAS 573-58-0, CR), an azoic dye that inhibits amyloid deposition, and some newly synthesized derivatives, more lipophilic and less toxic, were tested for their anti-prionic activity, in different experimental models. Cell-free experiments using the synthetic peptide PrP 106-126, homologous to amino acid residues 106-126 of the human PrP, were run to determine the anti amyloidogenic properties of some of the molecules. Peptide solutions containing each compound were incubated at 37 degrees C, for increasing times, to analyse the kinetics of aggregation of PrP 106-126 peptide. After incubation, the amount of non-aggregated peptide was measured by RP-HPLC. While CR enhanced the amyloidogenicity of PrP 106-126, derivatives "1a" and "1b" both showed the opposite behaviour, reducing aggregation by 15-20%. In other experiments using electron microscopy PrP 106-126 was assayed with the same molecules to assess the number and size of fibrils formed. CR showed its typical interaction, producing amyloid aggregates, "1a" did not interfere with fibril formation, while "1b" seemed to partially affect the structure of PrP 106-126 fibrils. Using a different cell-free model, it was investigated whether CR derivatives could reverse the protease-resistant PrPres, extracted from Syrian hamster infected brain, into the normal protease sensitive PrPsen. Samples containing fixed amounts of PrPres were incubated at 37 degrees C for 1 h with all the newly synthesized molecules, at concentrations ranging from 50 micrograms/mL to 750 micrograms/mL. After treatment with proteinase K, half of each sample was incubated with 3 mol/L guanidine thiocyanate in order to exclude over stabilisation of the PrPres aggregates already observed with CR. The remaining amount of PrPres was assessed by Enhanced Chemoluminescence (ECL) Western blotting analysis. None of the compounds induced the reversion of PrPres to PrPsen; nevertheless, 6 of the 8 molecules interacted with PrPres molecules, over stabilising the PrPres aggregates, from this aspect being similar to CR in activity. Finally, the inhibition of the generation of PrPres in the S12 clone of a mouse neuroblastoma cell line (N2a S12), persistently infected by the mouse adapted Chandler strain of scrapple, was evaluated. Increasing amounts of CR, "1a" and "1b" were added to the culture medium at each cell passage. After various days of treatment, the cells were collected, lysed, and the amount of PrPres was assayed by ECL Western blotting after PK treatment. As expected, there was a decrease in pathological PrP expression starting from the 4th day of treatment, with 5 and 10 micrograms/mL CR; PrPres completely disappeared after respectively 10 and 14 days of treatment. "1a" was strongly effective after 3 days of treatment at 5 and 10 micrograms/mL, but it was also highly toxic; at the concentration of 1 microgram/mL, it had a mild inhibitory effect after 8 days. The reduction of PrPres was also evaluated by intracytoplasmic flow-cytometry immunofluorescence on CR- and "1a"-treated N2a S12 cells. CR induced a dose related decrease of PrP expression from day 3 to 13 of treatment. At the concentrations of 2 and 1.5 micrograms/mL "1a" also strongly affected the expression of PrP starting from the 3rd day of treatment until the end of the experiment (day 13). These results confirm the importance of using an integrated system, based on different experimental models, to obtain useful information on the mechanism of action of anti-prionic compounds. PMID- 14750497 TI - Provision of pharmacy services to underserved populations via remote dispensing and two-way videoconferencing. AB - A telepharmacy program aimed to resolve the dual problem of access to prescription drugs and pharmacists for low-income populations is described. The program was conducted at the Community Health Association of Spokane (CHAS), a federally qualified community health center, and initiated in January 2001. The program, known as Telepharmacy at CHAS, allows for the dispensing of low-cost medications through participation in the federal government's 340B program and use of remote dispensing and counseling via a two-way interactive videoconferencing system to patients at six urban and rural clinics. The regulatory and practical steps necessary to implement and maintain the program are also presented. Consecutive patients whose prescriptions were filled at the base site or remote-sites were invited to complete a pharmacy satisfaction questionnaire during a two-week period in March 2003. Over the two-week period, 93 patients seen at remote sites and 106 seen at the base site completed the questionnaire. Over 75% of patients seen at the remote sites were satisfied with their videoconference interactions with the pharmacist. Of the patients seen at the base site, 66% agreed or strongly agreed that they were satisfied with the time required to obtain medications and counseling. A high percentage of patients at both the base site (94%) and remote sites (63%) strongly agreed or agreed that they would have difficulty affording their medications without this program. Telepharmacy at CHAS has been well received by most of the patients and improved their access to medications and pharmacy services. PMID- 14750498 TI - Photon-noise effect on detection in coherent active images. AB - We analyze photon-noise effects on target detection performance in low-flux coherent active imagery systems. We show that when photon noise is expected, the performance of classical detection techniques designed for pure and fully developed speckle images can be improved with no increase in algorithm complexity. Furthermore, the mean photon number under which photon noise becomes sensitive is higher when the target and background mean values are unknown than in the idealized case, where they are assumed to be known, and when the reflectivity ratio between the target and the background is low. PMID- 14750499 TI - Adaptor-tagged competitive PCR: study of the mammalian nervous system. AB - Adaptor-tagged competitive PCR (ATAC-PCR) is an advanced form of quantitative competitive PCR, and enables high-throughput analysis of gene expression. We applied this technique to the postnatal cerebellar development. Data analysis with terms representing reported functions revealed a correlation between gene expression and functions. We also analyzed the cell death induced by extended polyglutamine, a model of a neurodegenerative disorder. We identified genes with expression patterns specific to the cell death, and evaluated their functions by in vitro transfection experiments. PMID- 14750501 TI - [More about a new professional designation]. PMID- 14750500 TI - [Time to pregnancy and lead exposure]. AB - OBJECTIVE: To determine the effects of lead exposure on the time elapsed to become pregnant. MATERIAL AND METHODS: The study population consisted of 142 women residing in Mexico City between 1997 and 2001, who were already participating in a study to evaluate effects of lead exposure on reproductive health. Measurements of lead in bone were performed when women were first admitted to the program. Information on lead exposure and other variables of interest was obtained through a questionnaire. Participants were followed up to assess the relationship between the time required to become pregnant and lead exposure. Statistical analysis consisted of Kaplan-Meier estimates and Cox proportional hazards models. RESULTS: Of the total number of women in the program, 42 got pregnant: 34 before the first year of follow-up, and 8 at a later date. The mean value for lead concentration in blood was 9.3 micrograms/dl. The mean values for lead concentration in patella and tibia were 16.0 y 11.0 micrograms Pb/g of bone, respectively. Survival analysis was performed and no differences were detected in blood lead levels and time to pregnancy in the first year. Nevertheless, in women with blood lead levels above 10.0 micrograms/dl, the likelihood of not achieving pregnancy was five times higher (95% confidence interval [CI] 0.05-0.56) after one year of follow-up compared with women with blood lead levels below 10.0 micrograms/dl. CONCLUSIONS: Exposure to high lead concentrations may be an important risk factor influencing the time period for a woman to get pregnant, especially in fertile women who have tried to get pregnant for more than a year. This paper is available too at: http://www.insp.mx/salud/index.html. PMID- 14750502 TI - An all-atom solution-equilibrated model for human extrinsic blood coagulation complex (sTF-VIIa-Xa): a protein-protein docking and molecular dynamics refinement study. AB - Tissue factor (TF)-bound factor (F)VIIa plays a critical role in activating FX, an event that rapidly results in blood coagulation. Despite recent advances in the structural information about soluble TF (sTF)-bound VIIa and Xa individually, the atomic details of the ternary complex are not known. As part of our long-term goal to provide a structural understanding of the extrinsic blood coagulation pathway, we built an all atom solution-equilibrated model of the human sTF-VIIa Xa ternary complex using protein-protein docking and molecular dynamics (MD) simulations. The starting structural coordinates of sTF-VIIa and Xa were derived from dynamically equilibrated solution structures. Due to the flexible nature of the light-chain of the Xa molecule, a three-stage docking approach was employed in which SP (Arg195-Lys448)/EGF2 (Arg86-Arg139), EGF1 (Asp46-Thr85) and GLA (Ala1 Lys45) domains were docked in a sequential manner. The rigid-body docking approach of the FTDOCK method in conjunction with filtering based on biochemical knowledge from experimental site-specific mutagenesis studies provided the strategy. The best complex obtained from the docking experiments was further refined using MD simulations for 3 ns in explicit water. In addition to explaining most of the known experimental site-specific mutagenesis data pertaining to sTF-VIIa, our model also characterizes likely enzyme-binding exosites on FVIIa and Xa that may be involved in the ternary complex formation. According to the equilibrated model, the 140s loop of VIIa serves as the key recognition motif for complex formation. Stable interactions occur between the FVIIa 140s loop and the FXa -strand B2 region near the sodium-binding domain, the 160 s loop and the N-terminal activation loop regions. The helical-hydrophobic stack region that connects the GLA and EGF1 domains of VIIa and Xa appears to play a potential role in the membrane binding region of the ternary complex. The proposed model may serve as a reasonable structural basis for understanding the exosite-mediated substrate recognition of sTF-VIIa and to advance understanding of the TFPI-mediated regulatory pathway of the extrinsic blood coagulation cascade. PMID- 14750503 TI - A peptide isolated from alpha B-crystallin is a novel and potent inhibitor of platelet aggregation via dual prevention of PAR-1 and GPIb/V/IX. AB - BACKGROUND: The ability of low-molecular-weight heat shock protein (HSP) to modulate thrombin-induced platelet aggregation has been investigated. OBJECTIVES: We examined the inhibitory effects on platelet aggregation of nine amino acid sequences isolated from HSP20 or alpha B-crystallin and their various derivatives. METHODS AND RESULTS: Platelet aggregation induced by various agonists was performed. These findings indicated that a peptide (Trp-Ile-Arg-Arg Pro-Phe-Phe-Pro-Phe) from alpha B-crystallin significantly inhibits platelet aggregation induced by thrombin, TRAP (a protease activated receptor-1 agonist) and botrocetin, ristocetin (a stimulator of the platelet glycoprotein Ib/V/IX-von Willebrand factor axis), but not a protease-activated receptor-4 agonist, collagen and ADP. The inhibitory activity against thrombin or botrocetin is mainly linked to Arg-Arg-Pro-Phe or Trp-Ile-Arg-Arg-Pro, respectively, among nine amino acids. Additionally, during in vivo experiments, Trp-Ile-Arg-Arg-Pro-Phe Phe-Pro-Phe shows a significant antithrombotic effect without marked bleeding. CONCLUSION: Our results provide the basis for a potential new aspect of antiplatelet compound for the therapy of thrombosis and cardiovascular disease. PMID- 14750504 TI - [Proceeding of 33rd annual meeting of the Japanese Society of Neuropsychopharmacology. Nara, Japan. October 9-10, 2003. Abstracts]. PMID- 14750505 TI - Re: Is this glaucoma? PMID- 14750506 TI - [Canadian Thoracic Society COPD guidelines: summary of highlights for family doctors]. PMID- 14750507 TI - The plan. PMID- 14750508 TI - Naltrexone and nicotine patch smoking cessation: a preliminary study. AB - This preliminary study evaluated the potential efficacy of 50 mg of opioid antagonist naltrexone or placebo in combination with nicotine replacement therapy for smoking cessation, in a small sample of 32 smokers using one to one and a half packs of cigarettes per day. Addition of naltrexone resulted in an increase in continuous abstinence rates (9/16 for naltrexone vs. 5/16 for placebo). Naltrexone reduced the likelihood of relapse among participants who smoked during the first week of treatment, reduced desire to smoke, and prevented weight gain following smoking cessation. These preliminary results suggest that naltrexone augmentation of nicotine patch therapy using procedures designed to optimize the concurrent use of these two medications may be beneficial for smoking cessation. PMID- 14750509 TI - Unfolded proteins. PMID- 14750510 TI - Setting limits on selection of adults for liver transplantation. PMID- 14750511 TI - Bioethics and the challenge of the post-consensus society. PMID- 14750512 TI - [Commentary on the "Pathogenesis of decomposition in infancy" by Drs. Odon Kerpel Fronius and Ferenc Varga, Orvosi Hetilap 1953, 94, 29-36]. PMID- 14750513 TI - Mortality trend of ovarian cancer in Japan: 1960-2000. PMID- 14750514 TI - A tribute to I. B. Bender. PMID- 14750515 TI - Sport medicine journal club. PMID- 14750516 TI - A unified nomenclature for Arabidopsis dynamin-related large GTPases based on homology and possible functions. PMID- 14750517 TI - Domains of expansin gene expression define growth regions in the shoot apex of tomato. AB - Expansins are members of a multigene family of extracellular proteins, which increase cell wall extensibility in vitro and thus are thought to be involved in cell expansion. The major significance of the presence of this large gene family may be that distinctly expressed genes can independently regulate cell expansion in place and time. Here we report on LeExp9, a new expansin gene from tomato, and compare its expression in the shoot tip with that of LeExp2 and LeExp18. LeExp18 gene is expressed in very young tissues of the tomato shoot apex and the transcript levels are upregulated in the incipient primordium. LeExp2 mRNA accumulated in more mature tissues and transcript levels correlated with cell elongation in the elongation zone. In situ hybridization experiments showed a uniform distribution of LeExp9 mRNA in submeristematic tissues. When gibberellin deficient mutant tomatoes that lacked elongation of the internodes were treated with gibberellin, the phenotypic rescue was correlated with an increase in LeExp9 and LeExp2, but not LeExp18 levels. We propose that the three expansins define three distinct growing zones in the shoot tip. In the meristem proper, gibberellin-independent LeExp18 mediates the cell expansion that accompanies cell division. In the submeristematic zone, LeExp9 mediates cell expansion at a time that cell division comes to a halt. LeExp9 expression requires gibberellin but the hormone is not normally limiting. Finally, LeExp2 mediates cell elongation in young stem tissue. LeExp2 expression is limited by the available gibberellin. These data suggest that regulation of cell wall extensibility is controlled, at least in part, by differential regulation of expansin genes. PMID- 14750518 TI - Identification of rice (Oryza sativa) proteins linked to the cyclin-mediated regulation of the cell cycle. AB - Yeast two-hybrid assays were used to identify rice proteins interacting with two rice cyclins and other proteins potentially involved in cell cycling. The DNA sequences encoding 119 protein fragments identified were then compared by BLAST against proteins in GenBank. The proteins found include myosin-like proteins, transcription factors, kinesins, centromere proteins and undefined proteins. Based on interactions with cyclins and other elements required for cycling, we believe the undefined proteins may be involved in associated cycling processes. The identification of proteins involved in cell cycle regulation in rice may allow for the control of agronomic traits involving plant growth or development. PMID- 14750519 TI - Chromatin-associated HMGA and HMGB proteins: versatile co-regulators of DNA dependent processes. AB - High-mobility-group (HMG) proteins are small and relatively abundant chromatin associated proteins, which act as architectural factors. In plants, two groups of chromosomal HMG proteins have been identified, namely the HMGA family, typically containing four A/T-hook DNA-binding motifs, and the HMGB family, containing a single HMG-box DNA-binding domain. The HMGA proteins are structurally flexible and bind A/T-rich DNA stretches. By orchestrating multiple protein-protein and protein-DNA interactions, they assist the formation of higher-order transcription factor complexes, regulating gene expression. The HMGB proteins bind DNA non sequence-specifically, but specifically recognise DNA structures. Due to their remarkable DNA bending activity, they can enhance the structural flexibility of DNA, facilitating the assembly of nucleoprotein structures that control various DNA-dependent processes such as transcription and recombination. PMID- 14750520 TI - Phragmoplastin dynamics: multiple forms, microtubule association and their roles in cell plate formation in plants. AB - We have characterized 4 of the 16 members of the family of dynamin-related proteins (DRP) in Arabidopsis. Three members, DRP1A (previously referred as ADL1), DRP1C and DRP1E, belong to the largest group of phragmoplastin-like proteins. DRP2A (ADL6) is one of the two members that contain a pleckstrin homology (PH) domain and a proline-rich (PR) motif, characteristics of animal dynamins. All four proteins interacted in yeast two-hybrid assays with phragmoplastin, and showed different patterns of localization at the forming cell plate during cytokinesis. GFP-tagged DRP1A and DRP1C proteins were found to be associated with the cytoskeleton in G1 phase of the cell cycle. The distribution pattern of DRP1A was sensitive to propyzamid and insensitive to cytochalasin D, suggesting that DRP1A is associated with microtubules and not actin filaments. The association of DRP1A with microtubules was confirmed in vitro by spin-down assays. A GTPase-defective phragmoplastin acted as a dominant negative mutant, reduced transport of vesicles to the cell plate and formed dense tubule-like structures in the cell plate. We propose that DRP1 proteins may provide an anchor for Golgi-derived vesicles to attach to microtubules, which in turn direct the vesicles to the forming cell plate during cytokinesis. Whereas the DRP1 subfamily members are involved in tubulization of membranes, DRP2 may be involved in endocytosis and membrane recycling via clathrin-coated vesicles. PMID- 14750521 TI - ABI3 mediates expression of the peroxiredoxin antioxidant AtPER1 gene and induction by oxidative stress. AB - The peroxiredoxin antioxidant gene AtPER1 has been considered to be specifically expressed in the embryo and aleurone layer during maturation and desiccation stages of development, and in the mature seed, typically for late embryogenesis abundant (lea) transcripts. In the abscisic acid-insensitive abi3-1 mutant, the AtPER1 transcript level is strongly reduced, suggesting ABI3 to be a prime regulator of AtPER1. We have studied the expression pattern and regulation of AtPER1 with a series of nine promoter::GUS constructs with deletions and/or mutations in putative regulatory elements. Arabidopsis lines harbouring these constructs revealed AtPER1 promoter activity in the endosperm, especially the chalazal cyst, already when the embryo is in the late globular stage, in the embryo from the late torpedo stage, and in distinct cells of unfertilized and fertilized ovules. Early expression seems to be dependent on a putative antioxidant-responsive promoter element (ARE), while from the bent cotyledon stage endosperm and embryo expression is dependent on an ABA-responsive element (ABRE) likely to bind ABI5. The shortest promoter fragment (113 bp), devoid of ARE, ABRE and without an intact RY/Sph element thought to bind ABI3 did not drive GUS expression. The AtPER1::GUS construct also revealed expression in cotyledons, meristems and stem branching points. In general, seed and vegetative expression coincided with the expression pattern of ABI3. In plants ectopically expressing ABI3, AtPER1::GUS expression was found in true leaves, and AtPER1 could be induced by exogenous ABA and oxidative stress (H2O2 and hydroquinone). ABI3 mediated oxidative stress induction was dependent on the presence of an intact ARE element. PMID- 14750522 TI - Methylation of the exon/intron region in the Ubi1 promoter complex correlates with transgene silencing in barley. AB - Methylation of plant promoters is often associated with transcriptional gene silencing, while methylation of the transcribed region is associated with post transcriptional gene silencing. Promoter complexes that include the first untranslated exon and intron, such as maize ubiquitin1 and rice actin1, have been widely used in monocot transformation because they support higher levels of transient transgene expression than the core promoter does. However, persistent problems with transgene silencing driven by these promoter complexes brought up a troubling question: were higher initial levels of transgene expression at the expense of long-term expression stability? Here we report that methylation of an untranslated exon and intron in the promoter complex is correlated with transcriptional transgene silencing in barley. Two sibling sublines, designated T(3)30 and T(3)31, were identified in a homozygous T3 population from a single transgenic parental line. In the T6 generation, all progeny of one subline, T(3)30, expressed ubiquitin-driven bar and uidA, but both transgenes were transcriptionally silenced in the other subline, T(3)31. Although the structure of the transgene locus is complex, no differences in the physical structure or location of the locus were detected between the two sublines. Transcriptional transgene silencing in T(3)31 correlated with two molecular events: methylation of the first untranslated exon and 5' end of the intron of the promoter complex, and condensation of the chromatin in regions containing transgenes. Passage of the non-silenced subline through in vitro culture recreated the silenced phenotype of T(3)31 and the molecular events leading to its silencing. PMID- 14750523 TI - Characterization of the targeting signal of dual-targeted pea glutathione reductase. AB - We investigated the dual targeting signal of pea glutathione reductase (GR) that had been previously shown to be capable of targeting the passenger protein phosphinothricin acetyl transferase to mitochondria and chloroplasts in vivo. We confirmed that GR was imported into mitochondria and chloroplasts in vitro. Rupture of the outer mitochondrial membrane after the import assay indicated that GR was imported into both the intermembrane space and the matrix. Changing positive and hydrophobic residues in the targeting signal we investigated if dual targeting of GR was due to an overlapping or separate signal. Overall single mutations had a greater effect on mitochondrial import compared to chloroplasts, especially those on positive residues. Precursors containing both positive and hydrophobic residue mutations (double mutants) indicated that there might be some redundancy in targeting information for chloroplastic import as double mutants had a greater effect than predicted from the single mutants. Fusion of the targeting signal to the green fluorescent protein (GFP) followed by transient transformation indicated that this signal was only capable of targeting this passenger protein to plastids. Additionally, fusion of the complete coding sequence of GR to GFP also resulted in an exclusive chloroplastic localization. Mutations in the targeting signal that reduced import into plastids in vitro also displayed altered patterns of GFP localizations in vivo. These results indicate that some residues in the signal for dual localisation of GR play a role in both mitochondrial and chloroplastic import, and thus the signal is overlapping. PMID- 14750524 TI - The OsFOR1 gene encodes a polygalacturonase-inhibiting protein (PGIP) that regulates floral organ number in rice. AB - We have isolated a cDNA clone, OsFOR1, from the immature panicles of rice. The OsFOR1 (Oryza sativa floral organ regulator 1) gene encodes a protein that contains a leucine-rich repeat (LRR) domain. This domain comprises 10 tandem repeats of a canonical 24-amino acid LRR sequence. The structure and the number of LRRs for OsFOR1 are similar to those of polygalacturonase-inhibiting proteins (PGIPs) from various other plant species. Moreover, the OsFOR1 recombinant protein, when fused to maltose-binding protein (MBP), shows PGIP activity against the Aspergillus niger polygalacturonase. OsFOR1 is highly expressed in the calli and immature and mature panicles, while detectable at only low levels in seedling roots and mature stems. In situ hybridization experiments showed the transcripts of OsFOR1 are present in young spikelet primordia and in almost all of the young floral organs. Transgenic approaches were used to study in vivo functioning. Antisense expression of OsFOR1 resulted in an increase in the numbers of floral organs, including the stamen, carpel, palea/lemma, stigma, and lodicule. OsFOR1 transcript was not detected in the frizzy panicle mutant, which is defective in its spikelet formation but normal in inflorescence-meristem initiation and maintenance. Therefore, we suggest that OsFOR1 plays a role in the formation and/or maintenance of floral organ primordia. PMID- 14750525 TI - Expression and RNA binding properties of the chloroplast ribosomal protein S1 from Chlamydomonas reinhardtii. AB - The gene encoding the chloroplast ribosomal protein S1 from Chlamydomonas reinhardtii, CreS1, was cloned and the RNA binding properties and the expression patterns were studied. Gel-shift analysis revealed that CreS1 binds AU-rich 5' untranslated regions (5'-UTR) of chloroplast mRNAs with higher affinity than the corresponding sequence of a GC-rich nuclear transcript. The binding affinity of CreS1 for a mutant form of the psbD 5'-UTR with a deletion of a U-rich stretch that is required for translation decreases 4-fold as compared to the wild-type 5' UTR. Our results suggest that CreS1 protein interacts with U-rich sequences. Most of CreS1 is bound to high-molecular-weight complexes which co-migrate with the 30S small ribosomal subunit, and only a small fraction of CreS1 exists in its free form. CreS1 is localized mainly to the chloroplast stroma albeit a significant fraction is associated with chloroplast membranes. The results suggest that most of CreS1 is associated with the 30S ribosomal subunit throughout the translation process. Upon a shift of cells from the dark to the light, the mRNA levels of CreS1 and Psrp-7, both components of the 30S ribosomal subunit, increase transiently and return to the dark levels after 8 h. However, during this dark-to-light transition the levels of CreS1 and of other components of the 30S subunit remain the same suggesting that either protein synthesis or degradation is regulated. The possible implications of these findings are discussed. PMID- 14750527 TI - Highly abundant pea LTR retrotransposon Ogre is constitutively transcribed and partially spliced. AB - We have isolated and characterized a novel giant retroelement, named Ogre, which is over 22 kb long and makes up at least 5% of the pea (Pisum sativum L.) genome. This element can be classified as a Ty3/gypsy-like LTR retrotransposon based on the presence of long terminal repeats (LTRs) and the order of the domains coding for typical retrotransposon proteins. In addition to its extreme length, it has several features which make it unique among the retroelements described so far: (1) the sequences coding for gag and prot proteins are separated from the rt/rh int domains by several stop codons; (2) the region containing these stop codons is removed from the element transcripts by splicing which results in reconstitution of the complete gag-pol coding sequence; (3) only a part of the transcripts is spliced which probably determines the ratio of translated proteins; (4) the element contains an extra ORF located upstream the gag-pol coding sequences, potentially coding for a protein of 546-562 amino acids with unknown function. The transcriptional activity of the Ogre elements has been detected in all organs tested (leaves, roots, flowers) as well as in wounded leaves and protoplasts. Considering this retroelement's constitutive expression and observed high mutual similarity of the element genomic sequences, it is possible to speculate about its recent amplification in the genomes of pea and other legume plants. PMID- 14750526 TI - Molecular characterization of Brassica napus NAC domain transcriptional activators induced in response to biotic and abiotic stress. AB - Subtractive expressed sequence tag analysis and screening of cDNA libraries derived from Brassica napus leaves subjected to mechanical wounding, flea beetle feeding or cold temperatures revealed eight genes encoding NAC-domain transcription factors. The genes were found to be differentially regulated in response to biotic and abiotic stresses including wounding, insect feeding, Sclerotinia sclerotiorum infection, cold shock and dehydration. Five BnNAC proteins were orthologous to Arabidopsis thaliana ATAF1 or ATAF2 and gave rise to developmental abnormalities similar to the A. thaliana nam and cuc mutants when expressed ectopically in A. thaliana. Transgenic lines expressing BnNAC14, exhibited large leaves, thickened stems and hyper-developed lateral root systems similar to that observed with A. thaliana NAC1, but also were delayed in bolting and lacked an apical dominant tap root. Several of the BnNAC proteins were capable of activating gene expression in yeast and recognized an element within the CaMV35S promoter. A yeast two-hybrid screen revealed that BnNAC14 interacted with other select BnNAC proteins in vitro and identified an additional BnNAC gene, BnNAC485. The protein interaction and transcriptional activation domains were mapped by deletion analysis. PMID- 14750528 TI - Tomato EF-Ts(mt), a functional mitochondrial translation elongation factor from higher plants. AB - Ethylene-induced ripening in tomato (Lycopersicon esculentum) resulted in the accumulation of a transcript designated LeEF-Ts(mt) that encodes a protein with significant homology to bacterial Ts translational elongation factor (EF-Ts). Transient expression in tobacco and sunflower protoplasts of full-length and truncated LeEF-Ts(mt)-GFP fusion constructs and confocal microscopy observations clearly demonstrated the targeting of LeEF-Ts(mt) to mitochondria and not to chloroplasts and the requirement for a signal peptide for the proper sorting of the protein. Escherichia coli recombinant LeEF-Ts(mt) co-eluted from Ni-NTA resins with a protein corresponding to the molecular weight of the elongation factor EF-Tu of E. coli, indicating an interaction with bacterial EF-Tu. Increasing the GDP concentration in the extraction buffer reduced the amount of EF-Tu in the purified LeEF-Ts(mt) fraction. The purified LeEF-Ts(mt) stimulated the poly(U)-directed polymerization of phenylalanine 10-fold in the presence of EF-Tu. Furthermore, LeEF-Ts(mt) was capable of catalysing the nucleotide exchange reaction with E. coli EF-Tu. Altogether, these data demonstrate that LeEF-Ts(mt) encodes a functional mitochondrial EF-Ts. LeEF-Ts(mt) represents the first mitochondrial elongation factor to be isolated and functionally characterized in higher plants. PMID- 14750529 TI - Occupation and bladder cancer among men in Western Europe. AB - OBJECTIVES: We examined which occupations and industries are currently at high risk for bladder cancer in men. METHODS: We combined data from 11 case-control studies conducted between 1976-1996 in six European countries. The study comprised 3346 incident cases and 6840 controls, aged 30-79 years. Lifetime occupational and smoking histories were examined using common coding. RESULTS: Odds ratios for eight a priori defined high-risk occupations were low, and with the exception of metal workers and machinists (OR = 1.16, 95% CI = 1.02-1.32), were not statistically significant. Higher risks were observed for specific categories of painters, metal, textile and electrical workers, for miners, transport operators, excavating-machine operators, and also for non-industrial workers such as concierges and janitors. Industries entailing a high risk included salt mining, manufacture of carpets, paints, plastics and industrial chemicals. An increased risk was found for exposure to PAHs (OR for highest exposure tertile = 1.23, 95% CI = 1.07-1.4). The risk attributable to occupation ranged from 4.2 to 7.4%, with an estimated 4.3% for exposure to PAHs. CONCLUSIONS: Metal workers, machinists, transport equipment operators and miners are among the major occupations contributing to occupational bladder cancer in men in Western Europe. In this population one in 10 to one in 20 cancers of the bladder can be attributed to occupation. PMID- 14750530 TI - Association between non-steroidal anti-inflammatory drugs (NSAIDs) and invasive breast cancer and carcinoma in situ of the breast. AB - OBJECTIVE: Non-steroidal anti-inflammatory drugs (NSAIDs) have been consistently associated with reduced colon cancer risk. Associations with other cancer sites, particularly breast cancer, have been less consistent. This study's objective was to examine the relationship between use of non-prescription and prescription NSAIDs and breast cancer and carcinoma in situ of the breast. METHODS: We analyzed data from a population-based, case-control study conducted in North Carolina between 1996 and 2000 to examine the association between NSAID use and breast cancer. Logistic regression models were used to estimate odds ratios (ORs) and 95% confidence intervals (CI) separately for invasive breast cancer and carcinoma in situ of the breast. FINDINGS: NSAID use was inversely associated with invasive breast cancer (multivariable adjusted OR = 0.4; 95% CI: 0.3-0.6). Inverse associations were apparent for both prescription and non-prescription medications, and for occasional as well as regular use of NSAIDs. Inverse associations also were observed with carcinoma in situ, however the effects were somewhat weaker. INTERPRETATION: Our study adds to the accumulating, but somewhat inconsistent evidence that NSAIDs may reduce the risk of breast cancer. Future studies should work to define the most efficacious agents and determine whether genetic, lifestyle, or other characteristics influence their anti-carcinogenic properties. PMID- 14750531 TI - Population-density and county-level variation in breast cancer mortality rates among white women residing in the Northeastern and Southern United States. AB - OBJECTIVE: We assessed the contribution of variation in risk factor prevalence to population-density and county-level variation in breast cancer mortality rates. METHODS: In 1995 we collected risk factor information in a telephone interview of a random digit dialed sample of: (1) 1241 women from counties in the upper and lower tertiles of population density as of 1970 in the Northeast and South of the United States (Design A); (2) 2492 women from counties in the upper and lower tertiles of 1970-1979 breast cancer mortality rates in the four populations from Design A, and; (3) 276 women in Nassau County in New York State. We calculated 1990-94 mortality ratios (MRs) adjusted for breast cancer risk factors. RESULTS: The high/low population-density fully-adjusted MRs in women > or = 55 years were 1.01 (95% CI 0.9-1.2) and 1.00 (95% CI 0.8-1.2). The fully-adjusted MRs for high versus low mortality counties ranged from 0.95 (95% CI 0.8-1.2) to 1.29 (95% CI 1.0-1.6) in women > or = 55 years. CONCLUSIONS: Differences in risk factor prevalence explained higher rates in high-density versus low-density areas in older women. Modest elevations in the adjusted high/low breast cancer MRs among older women in certain groups of counties may reflect unidentified risk factors but more likely are due to chance. PMID- 14750533 TI - Survival from oral cancer in Mumbai (Bombay), India. AB - OBJECTIVE: Cancer survival information is available for only few populations in developing countries. Data on survival of oral cancer patients registered by the Bombay population-based cancer registry, India, during 1992-1994 is presented in this paper. METHODS: Vital status of the patients was established by matching against death certificates, reply paid postal enquiries, telephone enquiries, scrutiny of hospital records and house visits. Of the 1808 eligible cases for analysis, 1204 (66.6%) were dead; and 604 (33.4%) were alive at last follow-up, of whom 136 were lost to follow-up during the first 5-years after diagnosis. Observed and relative survival rates are presented and hazard ratios and corresponding 95% confidence intervals were estimated using Cox proportional hazard model. RESULTS: The overall 5-year observed and relative survival rates were 30.5% and 39.7%, respectively. Survival steadily declined with advancing age and advanced clinical stages. Five-year observed survival was 59.1% for localised cancer, 15.7% for cancers with regional extension and 1.6% for those with distant metastasis. Those with tongue, buccal mucosa and retromolar trigone cancers had poor survival rates. CONCLUSIONS: Our study shows that detecting oral cancer in early stages, when these are amenable to single modality therapies, offers the best chance of long-term survival. However, primary prevention by tobacco/alcohol control measures is important in the long-term perspective. PMID- 14750532 TI - Combined effect of oral contraceptive use and hormone replacement therapy on breast cancer risk in postmenopausal women. AB - OBJECTIVE: We examined breast cancer risk related to lifetime exposure to oral contraceptives (OCs) and hormone replacement therapy (HRT) in postmenopausal women. METHODS: The Women's Contraceptive and Reproductive Experiences (CARE) Study was a population-based case-control study that included 1847 postmenopausal women with incident invasive breast cancer, and 1932 control subjects, identified using random digit dialing. RESULTS: 45% of cases and 49% of controls used both OCs and HRT. OC users were not at increased risk regardless of subsequent HRT exposure. HRT users who had used OCs previously did not have a higher risk of breast cancer than women with no exposure to OCs. We observed a negative interaction (p-value: 0.032) of combined hormone replacement therapy (CHRT) and past OC use. The increase in risk with CHRT was stronger in women who had never used OCs in the past (odds ratio: 1.05; 95% confidence interval: 1.01-1.10 per year of exclusive CHRT use) than in women who had used OCs (odds ratio: 1.00; 95% confidence interval: 0.97-1.03). CONCLUSIONS: We found no indication that adverse effects of exposure to OCs or HRT appeared only in the presence of the other hormone or were exacerbated by exposure to the other hormone. PMID- 14750534 TI - Hypothesis: the act of taking a Papanicolaou smear reduces the prevalence of human papillomavirus infection: a potential impact on the risk of cervical cancer. AB - BACKGROUND: Based on data collected in a case-control study of hormonal contraceptive use and invasive cervical cancer among South African colored and black women, we have reported a five-fold reduction in incidence among women who in their lifetimes had previously received as few as two Papanicolaou (Pap) smears. Given the conditions prevailing in the community under study, a reduction in risk of this magnitude was surprising. We hypothesized that the act of performing a Pap smear may trigger an immune response to human papillomavirus (HPV) infection and thus reduce the risk of cervical cancer. METHODS: Among 1540 control women we compared prevalence rates of infection with specific HPVs known to cause cervical cancer among those who previously received 0, 1, 2 or > or = 3 Pap smears in their lifetimes. FINDINGS: Overall, 16% of the controls were HPV positive. Among women who received 0, 1, 2 and > or = 3 Pap smears the HPV positivity rates were 24, 17, 12, and 11%, respectively. Compared with never having received a smear, the confounder-adjusted relative risk estimates for the receipt of 1, 2, and > or = 3 smears were 0.7 (95% confidence interval, 0.5-1.0), 0.5 (0.3-0.8). and 0.5 (0.3-0.7), respectively (test for trend among Pap smear recipients: p = 0.04). The data were consistent within strata of age, parity, and race. INTERPRETATION: The data are compatible with the hypothesis that the minor trauma involved in taking a Pap smear may reduce the risk of cervical cancer by triggering an immune response to HPV infection. PMID- 14750535 TI - A prospective study of whole grains, fruits, vegetables and colon cancer risk. AB - OBJECTIVE: We examined the relation between whole grains, fruit, vegetables and dietary fiber and colon cancer risk in the prospective Cancer Prevention Study II Nutrition Cohort. METHODS: In 1992-1993, 62,609 men and 70,554 women completed questionnaires on medical history, diet and lifestyle behaviors. After exclusions, we confirmed 298 cases of incident colon cancer among men and 210 among women through August 31, 1997. RESULTS: Multivariate rate ratios (RR) and 95% confidence intervals (CI) for all dietary factors were null. However, a statistically non-significant 30% reduction in risk was observed for men with the highest vegetable intakes (RR = 0.69, CI = 0.47-1.03, top versus bottom quintile, p trend = 0.10). Men with very low (lowest tertile within the lowest quintile) intakes of vegetables and dietary fiber were at increased risk compared to those in the highest four quintiles of intake (vegetables RR = 1.79, CI = 1.22-2.61, p trend = 0.04, and fiber RR = 1.96, CI = 1.24-3.10, p trend = 0.006). Women with very low intakes of fruit were also at increased risk (RR = 1.86, CI = 1.18-2.94, p trend = 0.06). CONCLUSIONS: Higher intakes of plant foods or fiber were not related to lower risk of colon cancer. However, our data suggest that very low intakes of plant foods may increase risk, and that certain phytochemical subgroups may decrease risk. PMID- 14750536 TI - Palliative home care reduces time spent in hospital wards: a population-based study in the Tuscany Region, Italy. AB - OBJECTIVE: To estimate the reduction of time spent in hospital obtained through palliative home care. METHODS: Retrospective analysis of the hospital admissions occurred during the last year of life. Cancer patients who were resident in the province of Florence, were registered at the Tuscan Cancer Registry, and died during the year 1997 were included in the study. Three palliative home care services were operating in the study area in 1997. MAIN RESULTS: 3423 cancer patients died in the study area during 1997. 9.2% of them received palliative home care. Palliative home care was effective in reducing the utilization of hospital care during the last three months of life. A 25% reduction of the relative risk (CI: 34-14%) to have in-patient admissions and a 49% reduction of relative risk (CI: 52-47%) of spending days in the hospital during the last 3 months of life were estimated through multivariable regression models. CONCLUSIONS: Palliative home care was effective in reducing time spent in hospital during the last 3 months of life. PMID- 14750537 TI - Hormone replacement therapy in relation to survival in women diagnosed with colon cancer. AB - INTRODUCTION: Colorectal cancer is the second leading cause of cancer death among US men and women. A number of studies report that hormone replacement therapy (HRT) reduces the risk of colorectal cancer, but fewer studies have examined the relation between HRT use and survival after diagnosis and none examined this relation by anatomic location of the primary colon tumor. METHODS: Data from a cohort of 699 women 50-79 years of age diagnosed with colon cancer between 1980 and 1998 in a large Health Maintenance Organization based in Seattle, Washington were analyzed to examine the relation between HRT use prior to diagnosis and survival. Use of HRT was ascertained from a computerized pharmacy database. RESULTS: HRT use was associated with a 41% reduction in risk of death after adjustment for age, stage, and diagnosis year (p= 0.037). The association between HRT use and colon cancer survival was strongest among women with cancer in the distal colon (hazard rate ratio 0.33, 95% confidence interval, 95% CI: 0.13 0.83), while little or no association between HRT use and survival was observed among women with proximal tumors (HRR 0.78, 95% CI: 0.42-1.44). CONCLUSIONS: These findings suggest that HRT use is associated with improved survival from colon cancer. Future observational and laboratory studies should shed light on how hormones may be related to reduced incidence and improved prognosis. PMID- 14750538 TI - Changes in youth smoking participation in California in the 1990s. AB - OBJECTIVE: To identify which key transitions in the adolescent smoking uptake process changed during periods of increasing and decreasing adolescent smoking participation in California. METHODS: Age groups (12-14 and 15-17 years) from longitudinal surveys of California adolescents, conducted in 1993-1996 (prevalence increasing) and in 1996-1999 (prevalence declining), allowed comparisons of transition rates across these periods for: never smokers (committed and susceptible) to any smoking, experimenters to established smokers, and current established smokers to former smokers (15-17-year-olds only). Analyses adjusted for demographics and other baseline environmental influences on adolescents to smoke, with a variable for 'time period' included to test for differential period transition rates. RESULTS: Adjusted analyses indicated a time period effect (1993-1996 versus 1996-1999) on all transitions examined for 12-14 year-olds. For 15-17-year-olds, the time period effect was significant for the transition from committed never smoker to any smoking, but only marginally significant for experimenter to established smoker and from current established to former smoker. CONCLUSION: Adolescent smoking participation appeared to decline at all phases in the smoking uptake process, with generally larger declines in younger adolescents. Intensified public health measures to curb adolescent smoking beginning in the mid 1990s may have been responsible. PMID- 14750539 TI - Spontaneous clearance of high-titer serum HBV DNA and risk of hepatocellular carcinoma in a Chinese population. AB - Chronic hepatitis B virus (HBV) carriers with high-titer viremia (>10(5) virions/ml) are at increased risk for hepatocellular carcinoma (HCC). The aim of this study was to determine the relationship between clearance of high-titer viremia and subsequent risk of HCC. The study population was a prospective cohort of 114 adults from Haimen City, China, all HBV DNA(+) at study entry and followed for 797.8 person-years in total. During follow-up, 54 (47.4%) subjects spontaneously cleared high-titer viremia at least once. Of these, 27 were considered to have undergone stable seroconversion, 16 were considered unstable (12 reversions to HBV DNA positivity and 4 multiple clearances), and 11 did not have sufficient follow-up to determine stability. Of the 114 persons, 26 (22.8%) died during follow-up, 21 (18.4%) from HCC. Using Cox proportional hazards models, the RR of HCC death associated with seroconversion was 2.8 (95% CI = 1.1 7.4), controlling for age, sex, family HCC history, history of acute hepatitis, alcohol use and cigarette smoking. In conclusion, fluctuations of high-titer viremia may indicate increased hepatocellular damage and at least short-term increases in HCC risk. Long-term longitudinal studies are needed to clarify this relationship and its potential usefulness as a prognostic marker in chronic HBV infection. PMID- 14750540 TI - External validity in a population-based national prospective study--the Norwegian Women and Cancer Study (NOWAC). AB - OBJECTIVE: 'The Norwegian Women and Cancer Study' (NOWAC) was created as a national population-based cohort study by taking advantage of the existing population registers in Norway. Thus, the women in the NOWAC study should be representative for the entire female population in the corresponding age-groups. The aim of this paper was to study the external validity of the cohort as a necessary premise for the estimation of population attributable risk. METHODS: Between 1991 and 1997 women were sampled randomly from the national Central person registry in Norway. A total of 179,388 women were invited, of whom 102,443 can be included in the forthcoming follow-up analysis. The response-rates were around 60% in the age-groups 30-34 years till 55-59 years, while 44.7% responded among those aged 65-70 years. Information was collected by postal questionnaires. Follow-up information was based on linkage to national end-point registers with use of the unique national identification number given all Norwegians. RESULTS: Validation of the information on parity and education by linkage to national registers showed only minor differences for these factors between the responders and the total sample of women. A postal survey among non-responders indicated that the most important reasons for not returning the questionnaire were lack of time and concern about privacy. The results showed no differences in life-style factors between the original responders and the non-responders. No significant differences were found between the observed incidence rates for all cancer sites or cancer of the breast compared with national figures for the year 1999. CONCLUSION: The analysis revealed no major source of selection bias that could seriously invalidate the estimation of population attributable risk. PMID- 14750541 TI - Harvard launches a workshop series targeting the control of obesity in the US. AB - The Harvard Center for Cancer Prevention launched a workshop series to target the control of obesity in the United States. Under the direction of Professor Walter Willett, the series started on April 16 with a workshop titled 'Overweight/obesity and chronic disease: understanding and communicating the connection'. Several introductory lectures by distinguished experts in the field introduced the workshop participants to the key issues related to obesity control. During the second half of the meeting, participants broke out into small groups. Through lively discussions, each group developed a first set of key messages addressing preventive measures that need to be taken to effectively fight the obesity epidemic. Future workshops will help refine these messages with the ultimate goal of providing policy makers and communities with a tool for obesity control. PMID- 14750542 TI - Special issue: Bioethics & disability. PMID- 14750543 TI - Disability or end-of-life? Competing narratives in bioethics. AB - Bioethics, and indeed much ethical writing generally, makes its point through narratives. The religious parable no less than the medical teaching case uses a simple story to describe appropriate action or the application of a critical principle. While powerful, the telling story has limits. In this paper the authors describe a simple teaching case on "end-of-life" decision making that was ill received by its audience. The authors ill-received example, involving the disconnection of ventilation in a patient with ALS (Lou Gherig's Disease) was critiqued by audience members with long-term experience as ventilation users. In this case, the supposedly simple narrative of the presenters conflicted with the life histories of the audience. The lessons of this story, and the conflict that resulted, speak critically to the limits of simple teaching cases as well as the strengths of narrative analysis as a tool for the exploration of bioethical case histories. PMID- 14750544 TI - On the possibility and desirability of constructing a neutral conception of disability. AB - Disagreement about the proper attitude toward disability proliferates. Yet little attention has been paid to an important meta-question, namely, whether "disability" is an essentially contested concept. If so, recent debates between bioethicists and the disability movement leadership cannot be resolved. In this essay I identify some of the presumptions that make their encounters so contentious. Much more must happen, I argue, for any discussions about disability policy and politics to be productive. Progress depends on constructing a neutral conception of disability, one that neither devalues disability nor implies that persons with disabilities are inadequate. So, first, I clear away the conceptual underbrush that makes us think our idea of disability must be value-laden. Second, I sketch some constituents of, and constraints upon, a neutral notion of disability. PMID- 14750545 TI - Envisioning a kinder, gentler world: on recognition and remuneration for care workers. AB - In this paper, I argue that the status of those who take care of persons with disabilities, and persons with disabilities, are inextricably linked. That is, devaluing the status of one necessarily devalues that of the other. Persons with disabilities and those who help care for them must form an alliance to advance their common interests. This alliance can gain insight and inspiration from feminist thought insofar as caretaking is literally linked to problems of the representation of caretaking as "women's work,' and more philosophically, by borrowing from the toolbox of feminist social, political, and economic analyses. PMID- 14750546 TI - The ethics of receiving. AB - As a teacher and philosopher, Dr. Kate Lindemann has spent much of her professional life thinking about morality in human relationships. Critical analyses abound about the obligations and particular responsibilities of health care providers to patients, teachers to students, etc. Such analyses often emphasize the inherent inequality, and thus vulnerability, of those who are the recipients of care or knowledge. Though familiar with the ethics of care as a moral framework, Dr. Lindemann's perspectives on such relationships were profoundly affected and forever altered after acquiring a brain injury in 1998. The current manuscript describes how her views on caring acts as not only dynamic but reciprocal have been shaped by her experiences during rehabilitation and as a person now living with disability. PMID- 14750547 TI - Love and space in the nursing home. AB - Nursing homes and other institutions designed for persons with impairments are not, in fact, designed for persons with impairments. They are typically designed for the impairments, not the persons, and thereby become a part of the problem by reinforcing physical and cultural manifestations of the impairments. In the essay that follows, I describe an architectural design project in which students were asked to make changes to an existing nursing home for the persons who lived there. This requires not only becoming familiar with the spaces, but with the persons themselves and designing space to help articulate their voices and being. PMID- 14750548 TI - Disabilities and aging. AB - Both older persons and those who have disabilities can encounter discrimination when they seek medical care. Just as ageism and stereotypes about older persons may inappropriately limit medical care for the elderly, limits may be placed on medical care for those who are disabled simply because of the presence of a disability. At the same time death is the natural end of the lifespan for all individuals and there are situations when aggressive medical care is not indicated. It is not right to always insist on "doing everything" for a person even if that person may be at risk otherwise for discrimination. Using the example of the elderly, this paper examines the risks of discrimination and the dangers of overtreatment in caring for older persons and suggests parallels in the appropriate care of those who have disabilities. PMID- 14750549 TI - Should we re-examine the status of lymphocyte alloimmunization therapy for recurrent spontaneous abortion? AB - PROBLEM: In the human, lymphocyte alloimmunization immunotherapy (IT) has been proposed as a treatment for recurrent spontaneous abortion (RSA). This treatment has been the subject of debate for a long period of time. Recently, it has been proposed to extend such a treatment for implantation failure in humans, and I was asked to express my opinion on this topic. METHODS: I reviewed the evolution and theories and current paradigms in Reproductive Immunology and rationales proposed for IT. New discoveries show the complexity of implantation as a step by step developmental event, in mice and humans, and as such led me to re-examine paradigms currently evoked for extension of IT to implantation. Such a re examination obviously leads me to re-question the basis of IT for RSA itself. CONCLUSIONS: I conclude that the Th1/Th2 paradigm, evoked as the current basis for IT, and as useful as it has been to explain pregnancy, is no longer sufficient. It is especially insufficient to explain the process of implantation, which involves inflammatory molecules and cannot fit in such a scheme. It ensues that alloimmunization has no scientific rationale for the treatment of human implantation as a whole, and should not be applied broadly at such a stage of pregnancy. Furthermore, its use in RSA should be re-examined. PMID- 14750550 TI - Seven antiphospholipid antibodies in HIV-positive patients: correlation with clinical course and laboratory findings. AB - OBJECTIVE: To compare the clinical course of HIV-1-infected patients, their CD4+, CD8+ T lymphocytes, and viral loads (VL) with the levels of seven antiphospholipid antibodies (aPLs) before, during, and after the highly active antiretroviral therapy (HAART). PATIENTS AND METHODS: aPLs were examined in patients (20 men, 10 women, aged 12-64 years, median 33 years) from the AIDS center of Western Bohemia before the initiation of HAART, and two (23 patients), and five (20 patients) years later. Flow-cytometry was used for CD4+ and CD8+ T lymphocytes analysis, commercial kits were used for VL-measurements, and commercial enzyme-linked immunosorbent assay (ELISA) was used to determine serum levels of anti-beta2-glycoprotein I (GPI) of immunoglobulin G (IgG) and IgA isotypes, and anticardiolipin levels (ACA) of IgG and IgM isotypes. APLs screening also included L-alpha-phosphatidic (ph) acid, L-alpha phosphatidylethanolamine, L-alpha-phosphatidyl-DL-glycerol, L-alpha phosphatidylinositol, and L-alpha-phosphatidylserine of IgG and IgM autoantibodies. Statistical analysis was performed using cut-off levels for immunoglobulin-isotypes of aPLs using 3 S.D. or 95th percentile calculated using Statgraphics-software. RESULTS: In 14 of 21 patients treated by HAART an increased number of CD4+ T lymphocytes was detected, and in 14 of 21 patients VL decreased below detection threshold during the 5 years of observation. We did not observe correlations of aPLs with age, the initial low CD4+ and high number of CD8+ T lymphocytes, and the viremic levels over the entire observation period. We did not find the elevation of aPLs in 2 of 5 patients in stage C (AIDS). Ten of 11 HIV-positive homosexuals had positive aPLs, and the same result was seen in 7 of 10 patients infected through heterosexual intercourse. aPLs levels were significantly increased in 18 of 30 patients at the beginning of HAART. ACA IgG was elevated in 14 of 30 cases, IgG antibodies against L-alpha-ph-acid in 5 of 30, ph-ethanolamine in 10 of 30, ph-inositol in 9 of 30, and L-serine in 14 of 30, combined positivity of six aPLs together was detected in 10 HIV positive patients. Significantly decreased levels of aPLs because of HAART were found in eight patients. APLs were still present in only four patients after 5 years of the treatment. Abnormalities in blood clotting were not present in any of our patients. CONCLUSION: Results of screening for seven aPLs in HIV-positive patients suggest that HAART also positively influences the autoimmune response represented by aPLs levels, but individual differences in aPLs levels were observed. PMID- 14750551 TI - Cytokine secretion patterns of NK cells and macrophages in early human pregnancy decidua and blood: implications for suppressor macrophages in decidua. AB - PROBLEM: Local immune modulation has been shown to be of considerable importance for the maintenance of successful pregnancy. We have previously reported the secretion of interferon-gamma (IFN-gamma), interleukin-4 (IL-4) and IL-10 in human decidua from early normal pregnancy. The aim of this study was to investigate the cellular source of cytokine secretion in the decidua, and compare this to secretion patterns in peripheral blood. METHOD OF STUDY: Decidual tissue and peripheral blood was collected from 20 women undergoing surgical abortion during first trimester pregnancy. Monocytes/macrophages and NK cells were enriched by immunomagnetic cell separation and cytokine secretion was detected by enzyme-linked immunosorbent spot-forming cell assay. RESULTS: Decidual and peripheral monocytes/macrophages and NK cells spontaneously secrete IFN-gamma, IL 4 and IL-10. The number of IL-10 secreting cells was significantly higher in decidual macrophages compared with decidual non-monocytic cells as well as compared with blood monocytes/macrophages. These differences were not seen for IFN-gamma or IL-4. CONCLUSIONS: Our results indicate that decidual macrophages subserve important suppressive functions in the pregnant uterus. PMID- 14750552 TI - Differential gene expression of cytokine and cell surface molecules in T cell subpopulation derived from mammary gland secretion of cows. AB - PROBLEM: As T cell subpopulations in the mammary gland secretion (MGS) of cows dynamically vary through the lactation cycle, their functional analysis is important to understand the mammary immune responses. METHOD OF STUDY: T cell subpopulations were positively selected from MGS during lactation period and non lactation period (dry period) by a magnetic cell sorter. The messenger RNA (mRNA) expression of cytokine and cell surface molecules in the subpopulations stimulated with anti-CD3 was investigated using reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: CD4+ T cells from MGS significantly expressed mRNA of interferon (IFN)-gamma, interleukin (IL)-2, granulocyte-macrophage colony stimulating factor (GM-CSF), transforming growth factor (TGF)-beta, tumor necrosis factor (TNF)-alpha, IL-4, CD40 ligand (CD40L), Fas ligand (FasL) and IL 2 receptor (IL-2R) during dry period, and mRNA of IFN-gamma, IL-2 and TGF-beta during lactation period. Their expression during lactation period was always less than that during dry period. CD8+ T cells from MGS substantially expressed mRNA of IFN-gamma, IL-2, GM-CSF, TGF-beta, TNF-alpha, FasL and IL-2R during dry period and mRNA of IFN-gamma, GM-CSF, TGF-beta, TNF-alpha and c-kit during lactation period. The TGF-beta, TNF-alpha, c-kit and IL-2R mRNA expression of T cells in MGS during lactation period mostly depended on gammadelta T cells. Interestingly, c-kit mRNA was exclusively expressed in gammadelta T cells. CONCLUSIONS: The cytokine expression of T cells in MGS of cows depended on the T cell subpopulations. The present findings suggested that the activation of gammadelta T cells via c-kit receptor participated in the suppressed expression of cytokine mRNA in T cells during lactation period. PMID- 14750553 TI - Inhibition of human polymorphonuclear cell oxidative burst by 17-beta-estradiol and 2,3,7,8-tetrachlorodibenzo-p-dioxin. AB - PROBLEM: Polymorphonuclear cell (PMN) function may be directly influenced by 17 beta-estradiol and the endocrine disruptor, 2,3,7,8-tetrachloro-dibenzo-p-dioxin (TCDD). This may have significant consequences on PMN function within the female reproductive tract. This study evaluated the effects of 17-beta-estradiol and TCDD on PMN oxidative burst. METHOD OF STUDY: Peripheral blood PMN were isolated from normal male donors. Following treatment with 17-beta-estradiol, TCDD or both, PMN were stimulated with phorbol 12-myristate 13-acetate. Superoxide production was measured by lucigenin-enhanced chemiluminescence. RESULTS: Following 24-hr culture with either 17-beta-estradiol or TCDD, PMN superoxide production was significantly reduced, however, no such inhibition was observed when PMN were cultured with both estradiol and TCDD. Using antagonists, the estradiol and TCDD effects on PMN superoxide production was shown to be estrogen and aryl hydrocarbon receptor mediated. CONCLUSIONS: Estradiol and TCDD influence PMN oxidative burst through receptor mediated events. Such altered PMN function may have profound effects upon the normal endometrial cycle. PMID- 14750554 TI - Effect of prostatein, the major protein produced by the rat ventral prostate, on phagocytic cell functions. AB - PROBLEM: To determine whether prostatein, the major protein produced and secreted into the seminal fluid by the rat ventral prostate has any effect on the phagocytic cell functions in vitro. METHOD OF STUDY: Analysis was done by determining if purified prostatein added to cells obtained from the peritoneal cavity has any effect on their phagocytic and intracellular killing capacity. Also, we analyzed the effect of prostatein on the production of oxygen and nitrogen intermediates, measuring these metabolites by Nitroblue tetrazolium assay and by the Griess reaction respectively. RESULTS: Prostatein possess the ability to inhibit in vitro the phagocytic and killing properties of peritoneal rat leukocytes in a dose-dependent manner. The addition of a polyclonal antiserum against prostatein specifically blocks this inhibitory effect. Moreover, prostatein inhibits the production of oxygen and nitrogen intermediates by these cells. CONCLUSION: Regulation of the production of reactive oxygen species in the reproductive tract is extremely necessary to avoid their deleterious effects on the sperm motility and the fertilization process. We propose that prostatein, a protein supplied by an accessory gland like prostate, can inhibit the macrophage function, showing an important antioxidant effect. PMID- 14750555 TI - A digital method of sperm immobilization test: comparison to the conventional method. AB - Antisperm antibodies have been found in infertile patients and those causing immobilization of sperm are considered to be closely related to unexplained infertility. These antibodies are usually identified by a sperm immobilization test which involves counting motile sperm under microscope. This test is subjective as it relies on the judgement of the examiner with respect to sperm motility. In this study, we analyzed motile sperm by a digital method using Sperm Quality Analyzer. The results were compared with those obtained by the conventional method. We found that the two methods yielded identical results, with 14 of 66 samples tested being positive and 52 negative for sperm immobilizing antibodies. These results show that the digital method is objective and of value in the measurement of motile sperm in determination of sperm immobilizing antibodies. PMID- 14750556 TI - PCBs, hexachlorobenzene and DDE are not associated with recurrent miscarriage. AB - PROBLEM: A case-control study was designed to evaluate any associations between high exposure to polychlorinated biphenyls (PCB), hexachlorobenzene (HCB) and the 1,1,1,-trichloro-2,2-bis (p-chlorophenyl) ethane (DDT) metabolite 1,1-dichloro 2,2-bis (p-chlorophenyl) ethylene (DDE) and recurrent miscarriage and immunoendocrine abnormalities. METHODS OF STUDY: A total of 18 kinds of co-planer PCBs, HCB, DDE, natural killer cell (NK) activity, antiphospholipid antibodies, antinuclear antibody, prolactin, progesterone, thyroid-stimulating hormone (TSH) and free T4 were examined in 45 patients with a history of three or more (3-11) consecutive first-trimester miscarriages and 30 healthy women with no history of live birth and infertility. RESULTS: There were no differences in mean +/- S.D. values in serum samples for PCBs, HCB and DDE between patients and controls. Hypothyroidism, hyperprolactinemia, luteal phase defects, NK cell activity and the presence of autoantibodies were also not associated with levels of any of the compounds in the patients. CONCLUSION: PCBs, HCB and DDE are not associated with miscarriage and immunoendocrine abnormalities in patients with a history of recurrent miscarriage. PMID- 14750557 TI - Gamma/deltaT-cell subsets, NKG2A expression and apoptosis of Vdelta2+ T cells in pregnant women with or without risk of premature pregnancy termination. AB - PROBLEM: Potentially cytotoxic Vdelta2+ T lymphocytes recognize human leukocyte antigen-E on the trophoblast via their CD94/NKG2A receptors. This study aims at determing the percentage of gamma/delta T-cell subsets, their NKG2A and Annexin V positivity in peripheral blood of healthy pregnant women and women at risk of premature pregnancy termination. METHOD OF STUDY: Peripheral Vdelta2+ cells from healthy pregnant women and from women at risk of premature pregnancy termination were tested for the KIR NKG2A and Annexin V positivity by flow cytometry. RESULTS: The percentage of viable Vdelta2+ T cells was higher, that of Vdelta1+ T cells was lower in women at risk of premature pregnancy termination than in healthy pregnant women. The percentage of NKG2A + Vdelta2+ T cells was significantly lower in pregnant women at risk of premature pregnancy termination than in normal pregnancy. CONCLUSIONS: These data suggest the involvement of gamma/delta T lymphocytes in the pathogenesis of premature pregnancy termination. PMID- 14750558 TI - Technical note: fate and transport of jet fuel (JP-8) in soils with selected plants. AB - Remediation of soil and groundwater contaminated by leaking fuel storage tanks may be assisted by plants, although plant effects on abiotic and biotic removal processes remain unclear. The objectives of this study were to investigate abiotic and biotic removal of JP-8, a kerosene-based jet fuel, in soils with plants, and to determine the effects of plant-induced water movement. Loss of JP 8 in a dry-soil, control column was 25% after 5 months, primarily due to volatilization and gas-phase diffusion. By comparison, managed treatments with simulated surface spills averaged 86% mass reduction at 5 months, indicating an important contribution of biodegradation. Overall JP-8 mass reduction was similar in surface and subsurface-irrigated systems, indicating water content, not mode of water application, influences bioremediation in near-surface systems. The JP-8 concentration reductions in soil columns contaminated above a simulated watertable were 36% after 3 months and 50% after 12 months for vegetated columns compared to 26% and 34% in unplanted columns. Downward movement of JP-8 in unplanted columns was double that in planted columns. Near the groundwater table, JP-8 persists longer than near the soil surface. Plants promote upward movement of water and help draw spilled JP-8 to aerobic near-surface soil. PMID- 14750559 TI - Modeling jet fuel (JP-8) fate and transport in soils with plants. AB - Vegetation is often used to clean up soils and groundwater contaminated with organic contaminants. Plant-induced upward water movement may draw organic contaminants spilled near the watertable to the more aerated near-surface soil. The objective of this study was to develop and verify a 1-D model of fate and transport of JP-8, a kerosene-based jet fuel, in soil. The modeling approach considered the advective and dispersive transport of jet fuels dissolved in groundwater, which may undergo simple first-order decay or linear adsorption. The governing partial differential advection dispersion equation was solved in one dimension. Data from an experiment of fate and transport of JP-8 with plant induced upward water movement were used to verify the model. Simulated results with different scenarios described the experimental results well for different depths above the contaminated zone in both vegetated and unvegetated columns. Advection was the dominant mechanism near the contaminated zone and advection with retardation and decay was used to fit the data away from the contaminated zone. Results indicated that the soil water movement impacted the transport and concentration of JP-8 in the soil columns. This model can be used to simulate the fate of JP-8 associated with phytoremediation and evapotranspiration. PMID- 14750560 TI - Non-significance of rhizosphere degradation during phytoremediation of MTBE. AB - Methyl tertiary butyl ether (MTBE) is a gasoline additive associated with groundwater pollution at gas station sites. Previous research on poplar trees in hydroponic systems suggests that phytovolatilization is an effective mechanism for phytoremediation of MTBE (Rubin and Ramaswami, 2001), but the potential for microbial degradation of MTBE in the rhizosphere of trees had not been assessed. MTBE had largely been considered recalcitrant to microbial processes, but recent fieldwork suggests rapid biodegradation may occur in certain cases. This paper investigates the potential for rhizosphere degradation of MTBE at time frames relevant for phytoremediation. Three experiments were conducted at different levels of aggregation to examine possible degradation of MTBE by rhizosphere microorganisms that had been acclimated to low levels of MTBE for 6 weeks. MTBE soil die-away studies, conducted with both poplar trees and fescue grass, found no significant differences between MTBE concentration in vegetated and unvegetated soils over a two-week attenuation period. Closed chamber tests comparing hydroponic and rhizospheric poplar tree systems also showed essentially complete recovery of MTBE mass in both systems, suggesting an absence of degradation. Finally, rhizosphere microbes tested in aerated bioreactors were found to be thriving and metabolizing root materials, but did not show measurable degradation of MTBE. In all tests, the MTBE degradation product, Tert Butyl Alcohol (TBA), was not detected. The insignificance of MTBE degradation by rhizosphere microorganisms suggests that plant processes be the primary focus of further research on MTBE phytoremediation. PMID- 14750561 TI - Development of a hydroponic screening technique to assess heavy metal resistance in willow (Salix). AB - A nutrient thin film hydroponic system has been developed which allows rapid screening of willow (Salix) clones for their resistance to heavy metals, and hence their use in phytoremediation. Two clones known to be different in their resistance to heavy metals (Salix burjatica (Germany) and S. triandra x viminalis (Q83)), could be distinguished on the basis of leaf biomass, root biomass and stem height after 6 weeks. There were also differences in the uptake of heavy metals between the two clones. PMID- 14750562 TI - Screening of willow species for resistance to heavy metals: comparison of performance in a hydroponics system and field trials. AB - The aim of this study was to ascertain whether metal resistance in willow (Salix) clones grown in a hydroponics screening test correlated with data from the same clones grown independently in a field trial. If so, results from a short-term, glasshouse-based system could be extrapolated to the field, allowing rapid identification of willows suitable for planting in metal-contaminated substrates without necessitating longterm field trials. Principal Components Analysis was used to show groups of clones and to assess the relative importance of the parameters measured in both the hydroponics system and the field; including plant response factors such as increase in stem height, as well as metal concentrations in plant tissues. The clones tested fell into two distinct groups. Salix viminalis clones and the basket willow Black Maul (S. triandra) were less resistant to elevated concentrations of heavy metals than a group of hardier clones, including S. burjatica 'Germany,' S.x dasyclados, S. candida and S. spaethii. The more resistant clones produced more biomass in the glasshouse and field, and had higher metal concentrations in the wood. The less resistant clones had greater concentrations of Cu and Ni in the bark, and produced less biomass in the glasshouse and field. Significant relationships were found between the response of the same clones grown the in short-term glasshouse hydroponics system and in the field. PMID- 14750563 TI - Phenotypic characterization of microbes in the rhizosphere of Alyssum murale. AB - Metal hyperaccumulator plants like Alyssum murale are used for phytoremediation of Ni contaminated soils. Soil microorganisms are known to play an important role in nutrient acquisition for plants, however, little is known about the rhizosphere microorganisms of hyperaccumulators. Fresh and dry weight, and Ni and Fe concentrations in plant shoots were higher when A. murale was grown in non sterilized compared to sterilized soils. The analysis of microbial populations in the rhizosphere of A. murale and in bulk soils demonstrated that microbial numbers were affected by the presence of the plant. Significantly higher numbers of culturable actinomycetes, bacteria and fungi were found in the rhizosphere compared to bulk soil. A higher percent of Ni-resistant bacteria were also found in the rhizosphere compared to bulk soil. Percentage of acid producing bacteria was higher among the rhizosphere isolates compared to isolates from bulk soil. However, proportions of siderophore producing and phosphate solubilizing bacteria were not affected by the presence of the plant. We hypothesize that microbes in the rhizosphere of A. murale were capable of reducing soil pH leading to an increase in metal uptake by this hyperaccumulator. PMID- 14750564 TI - Influence of organic and inorganic soil amendments on plant growth in crude oil contaminated soil. AB - Phytoremediation can be a viable alternative to traditional, more costly remediation techniques. Three greenhouse studies were conducted to evaluate plant growth with different soil amendments in crude oil-contaminated soil. Growth of alfalfa (Medicago sativa L., cultivar: Riley), bermudagrass (Cynodon dactylon L., cultivar: Common), crabgrass (Digitaria sanguinalis, cultivar: Large), fescue (Lolium arundinaceum Schreb., cultivar: Kentucky 31), and ryegrass (Lolium multiflorum Lam., cultivar: Marshall) was determined in crude oil-contaminated soil amended with either inorganic fertilizer, hardwood sawdust, papermill sludge, broiler litter or unamended (control). In the first study, the addition of broiler litter reduced seed germination for ryegrass, fescue, and alfalfa. In the second study, bermudagrass grown in broiler litter-amended soil produced the most shoot biomass, bermudagrass produced the most root biomass, and crabgrass and bermudagrass produced the most root length. In the third study, soil amended with broiler litter resulted in the greatest reduction in gravimetric total petroleum hydrocarbon (TPH) levels across the six plant treatments following the 14-wk study. Ryegrass produced more root biomass than any other species when grown in inorganic fertilizer- or hardwood sawdust + inorganic fertilizer-amended soil. The studies demonstrated that soil amendments and plant species selection were important considerations for phytoremediation of crude oil-contaminated soil. PMID- 14750565 TI - Cobb's tufts: a rare cause of spontaneous hyphaema. AB - A 59-year-old hypertensive man presented to eye casualty complaining of blurring of vision in his right eye. There was no history of any trauma or coagulation defects. Examination revealed a 4 mm hyphaema in his right eye with no evidence of rubeosis or traumatic squeal. The intraocular pressure was 32 mm Hg. The patient was managed conservatively using topical steroids and antiglaucoma medication. A number of vascular tufts were identified on slit lamp biomicroscopy and confirmed on iris angiography PMID- 14750566 TI - Monocular visual loss in a patient undergoing cisplatin chemotherapy. AB - Chemotherapeutic agents used against tumours have a variety of toxic effects. We describe a case of acute blindness in the left eye of a patient after treatment with cisplatin for lung cancer. Both the clinical findings and the absence of either ocular or retrobulbar metastasis suggested that the condition was related to chemotoxicity. PMID- 14750567 TI - Modulation of intraocular pressure by unilateral and forced unilateral nostril breathing in young healthy human subjects. AB - PURPOSE: To determine the effects of unilateral right/left nostril breathing (URNB/ULNB) and forced unilateral right/left nostril breathing (FURNB/FULNB) on intraocular pressure (IOP) and to examine the differences in the IOP during the various phases of nasal cycle. METHODS: Young healthy volunteers of either sex aged between 19-24 years, participated in the sessions using URNB/ULNB (n = 52) and FURNB/FULNB (n = 28). The nostril dominance was calculated from signals recorded on the PowerLab equipment, representing pressure changes at the end of the nostrils during respiration. The IOP was measured with Tono-Pen. The subjects were divided into 4 groups viz. right nostril dominant (RND), left nostril dominant (LND), transitional right nostril dominant (TRND) and transitional left nostril dominant (TLND) groups. The IOP data 'before and after' URNB/ULNB or FURNB/FULNB were compared by using paired t-test. The baseline data of IOP between the groups were analysed by using independent samples t-test. RESULTS: The URNB decreased the IOP in the LND and TLND (p < 0.01) and also in the RND (p < 0.05) groups but not significantly in the TRND group. The ULNB decreased the IOP in the RND group (p < 0.01) only. The FURNB significantly reduced the IOP (p < 0.05) only in the LND and RND groups. The FULNB decreased the IOP but not significantly. The baseline IOP did not differ significantly between the LND, RND, TLND and TRND groups. CONCLUSION: The URNB/FURNB reduced the IOP, while ULNB/FULNB failed to increase the IOP significantly. It is suggested that the lowering of IOP by URNB indicated sympathetic stimulation. PMID- 14750568 TI - Pigmented striae of the anterior lens capsule and age-associated pigment dispersion of variable degree in a group of older African-Americans: an age, race, and gender matched study. AB - PURPOSE: To investigate pigmented striae of the anterior lens capsule in African Americans, a potential indicator of significant anterior segment pigment dispersion. METHODS: A group of 40 African-American subjects who exhibited pigmented lens striae (PLS) were identified from a non-referred, primary eye care population in Chicago, IL, USA. These subjects were then compared to an age, race, and gender matched control group relative to refractive error and the presence or absence of diabetes and hypertension. RESULTS: The PLS subjects (mean age = 65.4 +/- 8.8 years, range = 50-87 years) consisted of 36 females and 4 males. PLS were bilateral in 36 (85%) of the 40 subjects. Among the eyes with PLS, 21 (55%) of 38 right eyes and 22 (61%) of 36 left eyes also had significant corneal endothelial pigment dusting, commonly in the shape of a Krukenberg's spindle. Ten (25%) of the PLS subjects had either glaucoma or ocular hypertension (7 bilateral, 3 unilateral). The presence of trabecular meshwork pigment varied from minimal to heavy. The mean +/- SD (range) refractive error of the PLS right eyes was +1.61 +/- 1.43D (-1.50 to +5.00D) and +1.77 +/- 1.37D (-1.00 to +5.00D) for the left eyes. Based on these data, the PLS right eyes were +1.63D (Student's t, p = 0.0001; 95% CI = +0.82 to +2.44D) more hyperopic on average than the control right eyes, and the PLS left eyes were +1.77D (p = 0.0001; 95% CI = +0.92 to +2.63D) more hyperopic on average than the control left eyes. Trend analysis showed a gradually increasing likelihood of PLS with increasing magnitude of hyperopia in both eyes (Mantel-Haenszel chi-square, p = 0.001). Among PLS subjects, 24 (60%) of 40 were hypertensive and 9 (23%) of 40 were diabetic. However, these proportions were not significantly different (two-tailed Fisher's exact test; hypertension: p = 0.30; diabetes: p = 0.70) from the randomly selected controls. CONCLUSIONS: Among our African-American group, which consisted predominately of females >50 years of age, the likelihood of PLS increased with increasing hyperopic refractive error. This finding is consistent with the possibility that PLS may, in some circumstances, indicate a significant pigment dispersal process due to iris-lens rubbing that may be associated with crowding of anterior segment structures. Additional study is warranted to further assess the nature of PLS, their precise relationship with an age-related pigment dispersal process, and their true significance as a risk factor for development of glaucoma. PMID- 14750569 TI - Prophylaxis of acute posttraumatic bacterial endophthalmitis with or without combined intraocular antibiotics: a prospective, double-masked randomized pilot study. AB - PURPOSE: The effectiveness of an intraocular injection of combined gentamicin and clindamycin in the prevention of acute posttraumatic bacterial endophthalmitis following penetrating ocular injuries was evaluated in a prospective, double masked, randomized pilot study. METHODS: Sixty eyes of 60 patients with penetrating ocular injuries were treated at a tertiary care hospital. Following primary repair, the eyes were randomized in two groups. Group 1, the antibiotic injection group (cases), was given an intracameral or intravitreal injection of 0.1 mL antibiotic (40 microg gentamicin and 45 microg clindamycin). Group 2 (balanced saline solution [BSS] injection group [controls]) received intracameral or intravitreal injection of 0.1 mL BSS. All patients received standard prophylactic antibiotic therapy (systemic, subconjunctival, and topical). RESULT: Although the overall incidence of acute posttraumatic bacterial endophthalmitis was 6.6% (4 eyes), the results of three cultures were negative. All endophthalmitis cases occurred in the BSS injection group; however, there was no statistically significant difference between case and control groups (p = 0.11). The incidence rate for those with retained intraocular foreign bodies was 13.3% and for those without foreign bodies was 4.4%. No retinal toxicity was detected. CONCLUSION: Intraocular injection of gentamicin and clindamycin in addition to the other methods of prophylaxis may be an effective modality in the prevention of posttraumatic endophthalmitis. Early results suggest that these antibiotics may have a role as adjunct therapy to primary repair of injured globes without significant side effects at the dosage used. PMID- 14750570 TI - Use of optical coherence tomography (OCT) and indirect ophthalmoscopy in the diagnosis of macular edema in diabetic patients. AB - OBJECTIVES: To evaluate the use of Optical Coherence Tomography (OCT) in the diagnosis of macular edema (ME) in diabetic patients in comparison to indirect ophthalmoscopy (IO) and, in addition, to study the characteristics of these patients. METHODS: 165 patients were randomly selected to join the study in 1998. Ophthalmological, clinical and laboratory examinations were performed for all these patients. RESULTS: Diabetic retinopathy was identified in 143 eyes (44.7%) and ME in 58 (18.3% of the total and 40.5% of the patients with retinopathy). 82.7% (48) of the eyes with ME could be diagnosed with OCT, against 62.0% (36) with IO. Haemoglobin A1c was the only variable that showed a significant association with ME, when compared to control (p < 0.05). Retinopathy was associated with the presence of nephropathy (p = 0.01) and neuropathy (p = 0.001), but ME was not (NS for both). 68% of patients without ME had a visual acuity of more than 50%. CONCLUSIONS: OCT is a new method that can help the evaluation of ME in diabetic patients. It can be used not only to diagnose the lesion, but also to follow up the patients during treatment. High levels of haemoglobin A1c might be associated with the presence of ME. Diabetic complications (nephropathy and neuropathy) are associated with retinopathy but not with macular edema. PMID- 14750571 TI - Cataract surgery and transpupillary silicone oil removal through a single scleral tunnel incision under topical anesthesia; sutureless surgery. AB - The author evaluated the results of combined cataract extraction and transpupillary silicone oil removal through a single scleral tunnel incision, in eyes that had undergone pars plana vitrectomy with silicone oil tamponade. Twenty four of the 46 eyes were operated on under topical anesthesia with Blumenthal mode mini-nucleus manual extracapsular cataract extraction technique (mini-nuc ECCE), and silicone oil was removed passively through planned posterior capsulorhexis via the scleral tunnel, followed by endocapsular intraocular lens (IOL) implantation. The operation was completed without any suturing. The remaining 22 eyes were similarly operated on with the same cataract extraction technique, but in these cases silicone oil was classically aspirated actively through pars plana sclerotomies. Results were evaluated by visual acuity measurement, duration of operation, and complications. The transpupillary silicone oil removal group had significantly less vitreous hemorrhage (0- 31.8%) and posterior capsule opacification (0-36.4%). Also, the mean duration of the operation was significantly shorter in this group. There was no significant difference between the two groups with regard to postoperative recurrence of retinal detachment (12.5-18.1%) and visual acuity outcome. The combination of mini-nuc ECCE with transpupillary silicone oil removal compares favorably with the combination of silicone oil aspiration through pars plana sclerotomies. This combined technique allows the surgeon to perform the operation under topical anesthesia and no sutures are required. The intervention period is shorter and no posterior capsule opacification or vitreous hemorrhage develops. PMID- 14750572 TI - MRI artifact masquerading as orbital disease. AB - OBJECTIVES: To describe the diagnostic challenge associated with magnetic resonance imaging (MRI) artifact and to describe methods of minimizing this artifact. METHODS: Gadolinium enhancement was demonstrated to be artifact by using an inversion recovery pulse sequence technique. RESULTS: A patient with complaints of painless loss of vision also had radiologic abnormalities that led to subsequent referral to a tertiary care facility. Increased signals on the fat suppressed contrast enhanced MRI was demonstrated to be artifact by using an inversion recovery technique which produced a normal MRI of the orbit. CONCLUSIONS: Clinical confusion leading to unnecessary and costly evaluation may be avoided if one considers the possibility of artifact when reviewing magnetic resonance studies. When magnetic susceptibility is suspected, the use of an inversion recovery sequence can better delineate the true nature of the abnormality. PMID- 14750573 TI - Visual recovery patterns in children with Leber's hereditary optic neuropathy. AB - Three patients with Leber's hereditary optic neuropathy (LHON) showed spontaneous improvement in visual acuities after months of legal blindness. Two male patients with bilateral subacute visual loss were 14 years of age at presentation. The first male patient had a mitochondrial DNA mutation at nucleotide position 11778. The second male patient was found to be negative for the designated primary mutations (11778, 14484, 3460) and two of the secondary mutations (15257, 9804). The third patient was a 20-year-old female who presented with bilateral optic atrophy. She had been diagnosed as LHON and was found positive for the 3460 mutation when she was 15. These patients' pattern of visual recovery by developing small islands of normal vision within a central scotoma is characteristic in such rare cases of LHON. PMID- 14750574 TI - Regression of orbital myocysticercosis: an ultrasonographic study. PMID- 14750575 TI - The prognostic importance of detecting mild sensory impairment in leprosy: a randomized controlled trial (TRIPOD 2). AB - This study was designed to investigate whether leprosy patients diagnosed with mild sensory impairment have a better prognosis when treated with steroids than similarly impaired patients treated with placebo. A multi-centre, randomized, double-blind, placebo-controlled trial was conducted in Nepal and Bangladesh. Patients were eligible if they had a confirmed leprosy diagnosis, were between 15 and 50 years old, had mild sensory impairment of the ulnar or posterior tibial nerve of less than 6 months duration and did not require steroids for other reasons. 'Mild impairment' was defined as 'impaired on the Semmes-Weinstein monofilament test, but testing normal on the ballpen sensory test'. Subjects were randomized to either prednisolone treatment starting at 40 mg per day, tapering over 4 months, or placebo. Nerve function was monitored monthly. Any patient who deteriorated was taken out of the trial and was put on full-dose steroid treatment. Outcome assessment was done at 4, 6, 9 and 12 months from the start of the treatment. Outcome measures were the proportion of patients needing full-dose prednisolone and the Semmes-Weinstein sum scores. Each patient contributed only one nerve to the analysis. Seventy-five patients had nerves eligible for analysis, of whom 41 (55%) and 34 (45%) were allocated to the prednisolone and placebo arms, respectively. At 4 months, three patients in the prednisolone arm (7%) and six in the placebo arm (18%) had an outcome event requiring full dose steroids. At 12 months, these proportions had almost reversed, 11 (27%) and 6 (18%) in the treatment and placebo arms, respectively. In the latter group, 75% had recovered spontaneously after 12 months. Prednisolone treatment of sensory impairment of the ulnar and posterior tibial nerves detectable with the monofilament test, but not with the ballpen test, did not improve the long-term outcome in terms of recovery of touch sensibility, not did it reduce the risk of leprosy reactions or nerve function impairment beyond the initial 4-month treatment phase. Two unexpected main findings were the strong tendency of mild sensory impairment to recover spontaneously and the fact that patients with mild sensory impairment without any other signs or symptoms of reaction or nerve function impairment are relatively rare. PMID- 14750576 TI - Treatment with corticosteroids of long-standing nerve function impairment in leprosy: a randomized controlled trial (TRIPOD 3). AB - Some leprosy patients with long-standing nerve function impairment (NFI) appear to have responded favourably to treatment with corticosteroids. This study investigated whether patients with untreated NFI between 6 and 24 months duration and who are given standard regimen corticosteroid therapy, will have a better treatment outcome than a placebo group. A multicentre, randomized, double-blind placebo-controlled trial was conducted in Nepal and Bangladesh. Subjects were randomised to either prednisolone treatment starting at 40 mg/day, tapered by 5 mg every 2 weeks, and completed after 16 weeks, or placebo. Outcome assessments were at 4, 6, 9, and 12 months from the start of treatment. 92 MB patients on MDT were recruited, of whom 40 (45%) received prednisolone and 52 (55%) placebo treatment. No demonstrable additional improvement in nerve function, or in preventing further leprosy reaction events was seen in the prednisolone group. Overall, improvement of nerve function at 12 months was seen in about 50% of patients in both groups. Analysis of subgroups according to nerve (ulnar and posterior tibial), duration of NFI, and sensory and motor function, also did not reveal any differences between the treatment and placebo groups. There was however, indication of less deterioration of nerve function in the prednisolone group. Finally, there was no difference in the occurrence of adverse events between both groups. The trial confirms current practice not to treat long standing NFI with prednisolone. Spontaneous recovery of nerve function appears to be a common phenomenon in leprosy. Leprosy reactions and new NFI occurred in a third of the study group, emphasizing the need to keep patients under regular surveillance during MDT, and, where possible, after completion of MDT. PMID- 14750577 TI - Adverse events of standardized regimens of corticosteroids for prophylaxis and treatment of nerve function impairment in leprosy: results from the 'TRIPOD' trials. AB - Reactions in leprosy causing nerve function impairment (NFI) are increasingly treated with standardized regimens of corticosteroids, often under field conditions. Safety concerns led to an assessment of adverse events of corticosteroids, based on data of three trials studying prevention of NFI (the TRIPOD study). A multicentre, randomized, double-blind placebo-controlled trial was conducted in leprosy control programmes in Nepal and Bangladesh. Treatment was with prednisolone according to fixed schedules for 16 weeks, starting in one trial with 20 mg/day (prophylactic regimen: total dosage 1.96 g) and in the other two trials with 40 mg/day (therapeutic regimen: total dosage 2.52 g). Minor adverse events were defined as moon face, fungal infections, acne, and gastric pain requiring antacid. Major adverse events were defined as psychosis, peptic ulcer, glaucoma, cataract, diabetes and hypertension. Also, the occurrence of infected plantar, palmar, and corneal ulceration was monitored, together with occurrence of TB. Considering all three trials together, minor adverse events were observed in 130/815 patients (16%). Of these, 51/414 (12%) were in the placebo group and 79/401 (20%) in the prednisolone group. The relative risk for minor adverse events in the prednisolone group was 1.6 (P = 0.004). Adverse events with a significantly increased risk were acne, fungal infections and gastric pain. Major adverse events were observed in 15/815 patients (2%); 7/414 (2%) in the placebo group and 8/401 (2%) in the prednisolone group. No major adverse events had a significantly increased risk in the prednisolone arm of the trials. No cases of TB were observed in 300 patients who could be followed-up for 24 months. Standardized regimens of corticosteroids for both prophylaxis and treatment of reactions and NFI in leprosy under field conditions in developing countries are safe when a standard pre-treatment examination is performed, treatment for minor conditions can be carried out by field staff, referral for specialized medical care is possible, and sufficient follow-up is done during and after treatment. PMID- 14750578 TI - Treatment duration of reversal reaction: a reappraisal. Back to the past. AB - In this paper, earlier data on the length of prednisolone treatment are re examined. Based on those data and supported by the literature and the author's own observations, it can be concluded that type I leprosy reaction should be treated with prednisolone for a longer period than the 12 weeks, advised by the WHO. The paper also warns against silent nerve damage that may occur when prednisolone is discontinued early. PMID- 14750579 TI - Impairments and Hansen's disease control in Rondonia state, Amazon region of Brazil. AB - This retrospective study of impairments in a decentralized and integrated, routine Hansen's disease (HD) programme was done on a cohort of all new patients detected in Rondonia state from 1996 to 1999. It shows that the dynamics of impairments during treatment in Rondonia are similar to what has been published in other recent studies from Africa and Asia. Data about impairments at detection and at release from treatment (cure), the prescription of steroids, and epidemiological information are provided. Of the original 5350 new patients, 4230 patients (80%) completed multidrug therapy (MDT) and had complete data about their impairment status. At the start of treatment, 9% of the paucibacillary (PB) and 26% of the multibacillary (MB) patients had WHO grade 1 impairment. Three percent of the PB and 11% of the MB patients had visible deformities (WHO grade 2 impairment). Of the patients without impairments (grade 0) at the start of treatment, 5% of the PB and 20% of the MB patients developed impairments during treatment. Of the PB patients with a WHO impairment grade 1 at start of treatment, 34% improved and 6% got worse. Of the MB patients 34% improved and 12% became worse. In a separate study of patients from the 1997 intake, 17% of the PB and 58% of the MB patients were treated at least once with a course of steroids or thalidomide during MDT treatment. It is noted in the literature that the percentage of persons with recent nerve function impairment (NFI), nerve pain or tenderness and/or reaction reactions differs between projects. This may reflect real differences or may be caused by differences in routine monitoring and/or criteria and methods of treatment. The use of the WHO maximum score, particularly for the patients with grade 2, is not as sensitive to change as utilizing the summary of Eye, Hand and Foot (EHF) scores. If overall impairment figures are given, the proportions of MB patients may define the differences between projects, therefore it is important to analysis and present the results of PB and MB patients separately. The most simple (outcome) indicator to estimate the effectiveness of patient management would be the proportions of patients with impairment grade 0 at start of treatment who develop either grade 1 or 2 impairments during treatment. An additional (outcome) indicator could be the proportion of patients with impairment grade 1 at start of treatment who develop grade 2 impairments during treatment. Currently, no operational targets or acceptable level of performance for patient management have been set. This would be important to enable programme managers to determine if adequate patient education, treatment and follow up have been provided after the disease detection to prevent and/or minimize problems associated with the disease. The available evidence strongly suggests that reactions and impairments related to HD will continue to occur in large numbers, requiring the development of adequate infrastructures and sustainable services to detect and to manage problems associated with HD during and after MDT treatment. PMID- 14750580 TI - Delay in presentation, an indicator for nerve function status at registration and for treatment outcome--the experience of the Bangladesh Acute Nerve Damage Study cohort. AB - The objective of our research was to relate delay in presentation in the Bangladesh Acute Nerve Damage Study cohort to intake status and to treatment outcome. The Bangladesh Acute Nerve Damage Study (BANDS) is a prospective cohort study of 2664 consecutive newly registered patients at clinics run by the Danish Bangladesh Mission Leprosy (DBLM) project in Nilphamari, northern Bangladesh. The 1-year intake began in April 1995. Three-year follow-up for PB cases and 5 years for MB cases was completed in 2001. Delay in presentation in the BANDS cohort is associated with increased signs of nerve function impairment at registration. Individuals presenting with no nerve impairment and maintaining nerve function to the end of follow-up had the shortest mean delays. Individuals presenting with impairment that did not improve during follow-up had the longest mean delays. Discussion focuses on the value of setting a threshold value defining early presentation. Since the WHO Grade 2 disability rate effectively sanctions lengthy delays where there is no impairment, an indicator relating directly to delay is preferred as an indicator for good practice in leprosy control. PMID- 14750581 TI - Unexplained delayed nerve impairment in leprosy after treatment. AB - The objective of this study was to examine the clinical signs, symptoms and course of neuropathies in patients with leprosy who after treatment developed nerve impairment, not explained by relapse or reversal reactions. We searched the case-records of leprosy patients, seen between 1985 and 2002 at the department of dermatology at our centre. Included in the study were patients who had developed nerve impairment after treatment of leprosy in the absence of relapse, erythema nodosum leprosum, or reversal reactions, and who were referred to a neurologist. In these patients, we recorded age, onset of leprosy, type of leprosy, treatment of leprosy, signs and symptoms of delayed nerve impairment, results of electrophysiological studies, responses to treatment and course. Included were 14 patients, of whom eight had a (sub)acute multiple mononeuropathy (group I); and six had a slowly progressive multiple mononeuropathy (group II). Patients in group I had limited improvement of nerve impairment after treatment with corticosteroids, and recurrence of symptoms and signs (usually of the motor nerves) when corticosteroids were tapered off. Patients in group II had slowly progressive predominantly sensory nerve impairment. Initially, they had only subjective symptoms, after at least 3 years objective signs became detectable. These patients were not treated with immunosuppressants. Two groups of patients with unexplained delayed nerve impairment could be distinguished. One group had a multiple mononeuropathy resembling reversal reactions with insufficient response to corticosteroids. In these patients, more aggressive and prolonged immunosuppressive treatment should be considered. The aetiology for the neuropathy in the other group remains unclear and further investigations are needed to understand the pathogenesis before treatment recommendations can be given. PMID- 14750582 TI - WHO disability grading: operational definitions. AB - The WHO disability grading has been in use for many years. Its main use has been as an indicator for early case detection/reporting. More recently, the WHO grading has also been used as a change indicator of impairments for patients while on treatment. In such instances, the individual scores for eyes hand, and feet are added to obtain the so-called EHF sum score. A major drawback in the use of the grading system has been the lack of operational definitions of the descriptions for the grades. This may result in data and results of comparisons of data across programs and countries being flawed. The paper discusses the WHO grading in the light of its dual use: as an indicator for early case detection and as an indicator for change in impairments. The paper presents operational definitions for the grading options. PMID- 14750583 TI - Ulnar and median nerves in paucibacillary leprosy--a follow-up study of electrophysiological functions in patients before and after nerve trunk decompression. AB - Electrophysiological functions of ulnar and median nerves in paucibacillary leprosy patients were studied. Patients who showed deterioration of sensory motor functions in spite of steroid therapy were offered nerve decompression together with oral steroids. On periodic follow-up of those who opted for surgery, it was observed, in general, that NCV and amplitude remained reduced even though clinical recovery occurred. Only 80% recovery of electrophysiological functions was seen (as compared to control levels), even in cases that showed good results. Motor function recovered better than sensory function. Complete electrophysiological recovery, if it occurs at all, takes much longer than clinical recovery. PMID- 14750584 TI - Bilateral facial palsy in Hansen's disease. PMID- 14750585 TI - Chronic recurrent ENL, steroid dependent: long-term treatment with high dose clofazimine. PMID- 14750586 TI - The influence of age and gender on the driving patterns of older adults. AB - The purpose of this study was to investigate the influences of age and gender on the driving patterns of 300 older adults. Odds of driving less than every day increased significantly with age and female gender. However, no differences were found in the reduction of overall driving. Females were more likely than men to have stopped or reduced driving under certain adverse conditions and for elective purposes. The driving patterns of today's cohort of older females suggest that the gender gap may be narrowing. Social and cultural issues such as security, safety, and identity with driving may explain existing gender differences. PMID- 14750587 TI - On growing old in America: perceptions of the Okinawan war bride. AB - The unique perceptions of aging of Okinawan war brides are presented through interviews with 15 subjects. They voiced their thoughts about a range of topics, including available support systems, effects of acculturation, and their sense of security as they approach old age. Pride in culture and fears of lost family connections are expressed as these women speculate about the demands of aging in America. PMID- 14750588 TI - Knowledge and correlates of osteoporosis: a comparison of Israeli-Jewish and Israeli-Arab women. AB - The present study examined levels and correlates of knowledge about osteoporosis among 176 Israeli-Jewish (mean age = 55) and 80 Israeli-Arab (mean age = 51) women. Levels of knowledge about the disease were low among all women, especially regarding some of the risk factors. Knowledge and awareness about the disease were especially deficient among Arab women. Younger age and lower education were the main vulnerability factors among Jewish women, and lower desire to seek information from the medical establishment, higher religiosity, and the lack of extended medical insurance among Arab women. Educational programs, geared to the needs and capabilities of the different ethnic populations, should be encouraged. PMID- 14750589 TI - Aging happens: experiences of Swiss women living alone. AB - The purpose of this study was to give beginning insights into how aging is experienced by women living alone in Switzerland. A feminist methodology was used to gather and interpret 17 interviews conducted with a selected group of 9 older women living alone. Interviews were taped and transcribed for hermeneutic analysis; major themes were developed. Aging happens, Independence, Being vulnerable, Memory and aging, and If I had been a boy were the themes discovered. These experiences represent how aging is shaped by individual life courses, sociocultural conditions along with gender and class. PMID- 14750590 TI - Quality of life of older African American women in rural North Carolina. AB - PURPOSE: The purpose of this study was to investigate the relationship of activity to the quality of life of female African American elders. METHODS: Framed in the critical emancipatory paradigm, the study used qualitative research methodology, including the analysis of transcribed interviews and the critical review of relevant literature. RESULTS: Contributors to this study identified numerous past and present activities. The activities were bound together with the themes of "The Lord," Doing, Family, Environment, "I had to let it go," and "Just a part of life." DISCUSSION: Study results indicate that participants have enhanced their quality of life and emancipated themselves through engagement in a variety of activities that hold both real and symbolic meaning. PMID- 14750591 TI - Does financial self-efficacy explain gender differences in retirement saving strategies? AB - Research indicates that women remain less financially prepared for retirement than are men. Little research has examined the gender difference in use of retirement plans. The present research assessed the gender difference in use of employer-sponsored and private retirement plans, and sought to account for this difference. Social status and human capital factors, occupation and industry of employment, and sense of financial self-efficacy were expected to account for gender differences. Findings indicated gender has no relationship with use of private retirement plans. However, females were found to be less likely to use an employer-sponsored plan, and this was largely accounted for by gender differences in occupation. PMID- 14750592 TI - Understanding older women's health care concerns: a qualitative study. AB - BACKGROUND: Older women often have different physical and psychological health priorities compared to men, and health systems must strive to extend and improve health care delivery to meet older women's specific health care needs. The goal of this study was to obtain information from older women on how to improve health care services to best support their efforts to age successfully and receive optimal quality health care in later life. METHODS: Focus groups were conducted among women aged 65 or older recruited from the community in the Montreal, Quebec, area. A total of 36 women participated. The focus group sessions were audiotaped, and the transcripts of each session were analyzed for issues and themes emerging from the text. Content analysis using the framework approach was used to explore and understand the experience of the focus group participants. The data from the text were then coded according to the relevant and emergent ideas and concepts. RESULTS: Participants felt that their physical health care needs were being met, but that a number of issues relating to psychological health were inadequately addressed by health care professionals. The importance of feeling validated as active participants in a health care relationship, recognition of fears and anxieties associated with aging, and the need for information-sharing and education were all viewed as important health care priorities for older women. Time and accessibility were identified as the most significant barriers towards receiving optimal health care in later life. INTERPRETATION: The current health care system does not meet the global health care needs of older women. Health care leaders must recognize that success in program development and delivery for older women will require designing clinical programs that address both the physiological and psychosocial requirements of women. Only when women feel that they are being cared for in a comprehensive manner, one that includes attention to physical, psychological and emotional health, are we likely to be delivering health care that optimally promotes successful aging. PMID- 14750593 TI - Gene therapy of retinal dystrophies: achievements, challenges and prospects. AB - Early attempts at gene therapy of inherited retinal diseases by recombinant adenovirus-vectored gene replacement in laboratory animals met with moderate success but the effect was transient. Recently, emphasis has shifted to less toxic vectors, namely recombinant adeno-associated (rAAV) viruses. Ribozymes, targeted to the P23H rhodopsin mutation in transgenic rats, significantly reduced photoreceptor loss and slowed attenuation of the electroretinogram (ERG) for 8 months. By gene replacement, rAAV-based photoreceptor rescue has been achieved in the rds-/- mouse and has restored vision in dogs carrying a RPE65 gene mutation. Minigenes for neurotrophins delivered by rAAV have been effective in achieving structural rescue of photoreceptors in rodent models of dominant disease, although this has not always been accompanied by functional rescue. One of the current challenges is the application of ribozyme therapy for dominant mutations coupled with wild-type gene augmentation to overcome haploinsufficiency. Other animal models are currently being utilized for preclinical studies as well. Spontaneously mutated Irish Setters and rd mice offer excellent subjects for the therapy of recessive mutations as do the RPE65 knockout mouse and RCS (rdy) rat. With burgeoning preclinical successes, the future looks bright for the treatment and cure of inherited retinal diseases in human patients. PMID- 14750594 TI - Identifying retinal disease genes: how far have we come, how far do we have to go? AB - One of the great success stories in retinal disease (RD) research in the past decade has been identification of many of the genes and mutations causing inherited retinal degeneration. To date, more than 133 RD genes have been identified, encompassing many disorders such as retinitis pigmentosa, Leber congenital amaurosis, Usher syndrome and macular dystrophy. The most striking outcome of these findings is the exceptional heterogeneity involved: dozens of disease-causing mutations have been detected in most RD genes; mutations in many different genes can cause the same disease; and different mutations in the same gene may cause different diseases. Superimposed on this genetic heterogeneity is substantial clinical variability, even among family members with the same mutation. The RD genes involve many different pathways, and expression ranges from very limited (e.g. expressed in rod photoreceptors only) to ubiquitous. These findings raise several general questions--in addition to the extraordinary number of specific, biological problems revealed. What fraction of the patient population can now be accounted for by the known RD genes? How many more RD genes will be found, and how should we find them? Are we dealing with just a handful of disease mechanisms or are there many different routes to retinal degeneration? How will this extreme heterogeneity affect our ability to diagnose and treat patients? These questions are considered in this summary. PMID- 14750595 TI - Dominant cone and cone-rod dystrophies: functional analysis of mutations in retGC1 and GCAP1. AB - The regulation of cGMP levels is central to the normal process of phototransduction in both cone and rod photoreceptor cells. Two of the proteins involved in this process are the enzyme, retinal guanylate cyclase (retGC), and its activating protein (GCAP) through which activity is regulated via changes in cellular Ca2+ levels. Dominant cone-rod dystrophies arising from changes in retGC1 are essentially restricted to mutations in codon 838 and result in the replacement of a conserved arginine residue with either cysteine, histidine or serine. In all three cases, the effect of the substitution on the in vitro cyclase activity is a loss of Ca2+ sensitivity arising from an increased stability of the coiled-coil domain of the protein dimer and retention of cyclase activity. In contrast, mutations in the Ca2+-coordinating EF hands of GCAP1 result in dominant cone dystrophy; the consequences of these mutations is a reduced ability of the mutant protein to regulate retGC activity in response to changes in Ca2+ levels. Functionally therefore, the retGC2 and GCAP2 mutations are similar in reducing the feedback inhibition of Ca2+ on cyclase activity and thereby on cGMP levels in the photoreceptors. PMID- 14750596 TI - Isotretinoin treatment inhibits lipofuscin accumulation in a mouse model of recessive Stargardt's macular degeneration. AB - Recessive Stargardt's macular degeneration is an inherited blinding disease of children caused by mutations in the ABCR gene. The primary pathologic defect in Stargardt's discase is accumulation of toxic lipofuscin pigments such as N retinylidene-N-retinylethanolamine (A2E) in cells of the retinal pigment epithelium (RPE). This accumulation appears to be responsible for the photoreceptor death and severe visual loss in Stargardt's patients. Here, we tested a novel therapeutic strategy to inhibit lipofuscin accumulation in a mouse model of recessive Stargardt's disease. Isotretinoin (Accutane) has been shown to slow the synthesis of 11-cis-retinaldehyde (11cRAL) and regeneration of rhodopsin by inhibiting 11-cis-retinol dehydrogenase (11cRDH) in the visual cycle. Light activation of rhodopsin results in its release of all-trans-retinaldehyde (atRAL), which constitutes the first reactant in A2E biosynthesis. Accordingly, we tested the effects of isotretinoin on lipofuscin accumulation in abcr-/- knockout mice. Isotretinoin blocked the formation of A2E biochemically and the accumulation of lipofuscin pigments by electron microscopy. We observed no significant visual loss in treated abcr-/- mice by electroretinography. Isotretinoin also blocked the slower, age-dependent accumulation of lipofuscin in wild-type mice. These results corroborate the proposed mechanism of A2E biogenesis. Further, they suggest that treatment with isotretinoin may inhibit lipofuscin accumulation and thus delay the onset of visual loss in Stargardt's patients. Finally, the results suggest that isotretinoin may be an effective treatment for other forms of retinal or macular degeneration associated with lipofuscin accumulation. PMID- 14750597 TI - The expanding roles of ABCA4 and CRB1 in inherited blindness. AB - Mutations in the ABCA4 gene cause Stargardt disease (STGD), most cases with autosomal recessive (ar) cone-rod dystrophy (CRD), and some cases with atypical ar retinitis pigmentosa (arRP). We found compound heterozygous ABCA4 mutations in two unrelated patients with STGD and homozygous splice site mutations in their 2nd and 4th degree cousins with RP. Some ABCA4 mutations display strong founder effects. In Dutch and German STGD patients, the 768G > T mutation is present in 8% and 0.6% of ABCA4 alleles respectively. Vice versa, the complex L541P;A1038V allele is found in 70% of ABCA4 alleles in German STGD patients but absent in Dutch patients. As approximately 70% of ABCA4 mutations are known, a microarray based analysis of known ABCA4 gene variants allows routine DNA diagnostics in Caucasian patients. Mutations in the CRB1 gene underlie RP12, some cases with classic arRP, 55% of cases with RP and Coats-like exudative vasculopathy, and 13% of patients with Leber congenital amaurosis (LCA), rendering CRB1 a significant cause of autosomal recessive retinal dystrophy. Different combinations of mutations in ABCA4 or CRB1 can be correlated with disease severity, suggesting that small increments of protein activities in patients might have significant therapeutic effects. Mouse and Drosophila studies strongly suggest that both patient groups might benefit from reduced light exposure and therefore should be detected as early as possible using molecular techniques. PMID- 14750598 TI - What should a clinician know to be prepared for the advent of treatment of retinal dystrophies? AB - It is now evident that several forms of therapy have influenced inherited retinal degeneration in animals. These are gene therapy, cell transplantation, slowing of cell death using growth factors and pharmacological approaches. There are firm proposals to initiate gene therapy in the foreseeable future. For therapy to be successful a variety of attributes of disease must be established so that the full benefits of research can be realised in clinical practice. These can be considered as identification of the causative genes, knowledge of the disease mechanisms and detection of the therapeutic effect. PMID- 14750599 TI - Role of subunit assembly in autosomal dominant retinitis pigmentosa linked to mutations in peripherin 2. AB - Peripherin 2 is a photoreceptor-specific membrane protein implicated in outer segment disk morphogenesis and linked to various retinopathies including autosomal dominant retinitis pigmentosa (ADRP). Peripherin 2 and ROM1 assemble as a mixture of core noncovalent homomeric and heteromeric tetramers that further link together through disulfide bonds to form higher order oligomers. These complexes are critical for disk rim formation and outer segment structure through interaction with the cGMP-gated channel and other photoreceptor proteins. We have examined the role of subunit assembly in peripherin 2 targeting to disks, outer segment structure, and photoreceptor degeneration by examining molecular and cellular properties of peripherin 2 mutants in COS-1 cells and transgenic Xenopus laevis rod photoreceptors. Wild-type (WT) and the ADRP-linked P216L mutant were transported and incorporated into newly formed outer segment disks of transgenic X. laevis. The P216L mutant, however, induced progressive outer segment instability and photoreceptor degeneration possibly through the introduction of a new N-linked oligosaccharide chain. In contrast, the C214S and L185P disease linked, tetramerization-defective mutants, were retained in the inner segment, but did not affect outer segment structure or induce photoreceptor degeneration. Together, these results indicate that peripherin 2 mutations can cause ADRP either through a deficiency in WT peripherin 2 (C214S, 1.185P) or by a dominant negative effect on disk stability (P216L). PMID- 14750600 TI - The search for rod-dependent cone viability factors, secreted factors promoting cone viability. AB - During the last decade, numerous research reports have considerably improved our knowledge of the pathophysiology of retinal degenerations. Three non-mutually exclusive general areas dealing with therapeutic approaches have been proposed: gene therapy, pharmacology and retinal transplantations. The observation that cone photoreceptors, even those seemingly unaffected by any described anomaly, die secondarily to rod disappearance related to mutations expressed specifically in the latter, led us to study the interactions between these two photoreceptor populations to search for possible causal links between rod degeneration and cone death. These in vivo and in vitro studies suggest that paracrine interactions between both cell types exist and that rods are necessary for continued cone survival. We have developed a protocol that is used to evaluate the potential of all sequences in a retinal library to generate a protective effect on cones from cone-enriched cultures from chicken embryo. The protocol of expression cloning is a systematic approach aimed at screening all genes normally expressed by retina. Since the role of cones in visual perception is essential, pending the identification of the factors mediating these interactions underway, rod replacement by transplantation and/or neuroprotection by trophic factors or alternative pharmacological means appear as promising approaches for limiting secondary cone loss in currently untreatable blinding conditions. PMID- 14750601 TI - Studies on retinal and retinal pigment epithelial gene expression. AB - The 'completion' of the murine and human genomes and creation of high-density expressed sequence tag (EST) databases from multiple tissues and multiple species, coupled with the development of high-throughput expression profiling approaches such as microarrays and Serial Analysis of Gene Expression (SAGE), is making possible the in-depth analysis of gene expression patterns in health and disease to an extent that was not previously possible. Such new information is providing insight into normal function, and into how normal function is altered in disease. Efforts have begun, and are accelerating, in the application of expression profiling to the study of the retina and retinal pigment epithelium (RPE). In this chapter we will review progress in this area. We will also discuss technical issues that make expression studies of the RPE particularly challenging, and share our experience in methodological approaches to overcome these challenges. PMID- 14750602 TI - From disease genes to cellular pathways: a progress report. AB - Mutations in a large number of retinal and retinal pigment epithelium (RPE) expressed genes can lead to the degeneration of photoreceptors and consequently the loss of vision. The genetic and phenotypic heterogeneity of retinal dystrophies poses a complex problem with respect to rational development of therapeutic strategies. Delineation of physiological functions of disease genes and identification of pathways that lead to disease pathogenesis represent essential goals towards developing a systematic and global approach to gene-based treatments. We are interested in identifying cellular pathways that are involved in photoreceptor differentiation, function and degeneration. We are, therefore, generating comprehensive gene expression profiles of retina and RPE of humans and mice using both cDNA- and oligonucleotide-based (Affymetrix) microarrays. Because of the under-representation of retinal/RPE genes in the public databases, we have constructed several unamplified cDNA libraries and produced almost twenty thousand expressed sequence tags (ESTs) that are being printed onto glass slides ('I-Gene' microarrays). In this presentation, we will report the microarray analysis of the rodless (and cone-enhanced) retina from the Nrl-knockout mouse as a paradigm to initiate the identification of cellular pathways involved in photoreceptor differentiation and function. PMID- 14750603 TI - Prospects for gene therapy. AB - Inherited retinal disease, which includes conditions such as retinitis pigmentosa (RP), affects about 1/3000 of the population in the Western world. It is characterized by gradual loss of vision and results from mutations in any one of 60 or so different genes. There are currently no effective treatments, but many of the genes have now been identified and their functions elucidated, providing a major impetus to develop gene-based treatments. Many of the disease genes are photoreceptor- or retinal pigment epithelium (RPE) cell specific. Since adeno associated viral (AAV) vectors can be used for efficient gene transfer to these two cell types, we are developing AAV-mediated gene therapy approaches for inherited retinal degeneration using animal models that have defects in these cells. The retinal degeneration slow (rds or Prph2Rd2/Rd) mouse, a model of recessive RP, lacks a functional gene encoding peripherin 2, which is a photoreceptor-specific protein required for the formation of outer segment discs. We have previously demonstrated restoration of photoreceptor ultrastructure and function by AAV-mediated gene transfer of peripherin 2. We have now extended our assessment to central visual neuronal responses in order to show an improvement of central visual function. The Royal College of Surgeons (RCS) rat, provides another model of recessive RP. Here the defect is due to a defect in Mertk, a gene that is expressed in the RPE and encodes a receptor tyrosine kinase that is thought to be involved in the recognition and binding of outer segment debris. The gene defect results in the inability of the RPE to phagocytose the shed outer segments from photoreceptor cells. The resulting accumulation of debris between the RPE and the neuroretina leads to progressive loss of photoreceptor cells. AAV mediated delivery of Mertk to the RPE results in reduction of debris indicating that the phagocytosing function of the RPE is restored and delays the degeneration of the photoreceptor cells 3-4 months. Our results, along with those of other groups support the use of AAV vectors for the treatment of inherited retinal degeneration. PMID- 14750604 TI - Range of retinal diseases potentially treatable by AAV-vectored gene therapy. AB - Viable strategies for retinal gene therapy must be designed to cope with the genetic nature of the disease and/or the primary pathologic process responsible for retinal malfunction. For dominant gene defects the aim must be to destroy the presumably toxic gene product, for recessive gene defects the direct approach aims to provide a wild-type copy of the gene to the affected retinal cell type, and for diseases of either complex or unknown genetic origin, more general cell survival strategies that deal with preserving affected retinal cells are often the best and only option. Hence examples of each type of therapy will be briefly discussed in several animal models, including ribozyme therapy for autosomal dominant retinitis pigmentosa in the transgenic P23H opsin rat, beta-PDE gene augmentation therapy for autosomal recessive retinitis pigmentosa in the rd mouse, glial cell-derived neurotrophic factor (GDNF) gene therapy for autosomal dominant RP in the transgenic S334ter opsin rat and pigment epithelial cell derived neurotrophic factor (PEDF) gene therapy for neovascular retinal disease in rodents. Each employs a recombinant AAV vectored passenger gene controlled by one of several promoters supporting either photoreceptor-specific expression or more general retinal cell expression depending on the therapeutic requirements. PMID- 14750605 TI - Gene therapy for Leber congenital amaurosis. AB - Recent success in delivering vision to a canine model of a severe, early-onset blinding disease, Leber congenital amaurosis (LCA) (Acland et al 2001) demonstrates that adeno-associated virus serotype 2 (AAV2) is capable of delivering a corrective gene to the target retinal cells. Results of these studies indicate long-term rescue of vision as assessed by psychophysical, behavioural and molecular biological studies. Preliminary results of studies in progress are described and the implications of these results with respect to developing human clinical trials for LCA and for other retinal degenerative diseases are discussed. PMID- 14750606 TI - A superantigen hypothesis for the pathogenesis of chronic hyperplastic sinusitis with massive nasal polyposis. AB - BACKGROUND: The pathogenesis of chronic hyperplastic sinusitis with massive nasal polyposis is still an enigma; however, the molecular biology of this disease is beginning to become unraveled and the proinflammatory cytokines and the message and the product of these cytokines have all been identified in nasal polyps. However, the initial trigger that causes inflammation of the lateral wall of the nose to up-regulate lymphocytes and eosinophils is still unknown. METHODS: Thirteen patients with massive polyposis were studied. The mucus of the nasal cavities surrounding the nasal polyps was studied for both bacterial and fungal species. The lymphocytes of the nasal polyps were extracted and evaluated for the T-cell receptor, particularly, the variable beta region of this receptor. Enterotoxins (superantigens) of the bacteria were studied. Finally, the histopathology of nasal polyps was studied. RESULTS: Fifty-five percent of the patients had toxin-producing Staphylococcus aureus in the nasal mucus adjacent to the polyps. Three different enterotoxins were isolated, including Staphylococcus enterotoxin A, Staphylococcus enterotoxin B, and toxic shock syndrome toxin 1. The variable B specificity for these superantigens was identified also in the polyp lymphocyte T-cell receptor. CONCLUSION: A superantigen hypothesis for massive polyposis is suggested because the most common bacterial species found in the nasal mucus is Staphylococcus aureus. These bacteria produce enterotoxins in all of the cases studied and the corresponding variable beta region of the T-cell receptor also was up-regulated in the polyp lymphocytes in cases studied thus far. These data taken together suggest that the initial injury to the lateral wall of the nose may be the result of toxin-producing Staphylococci. Superantigens (enterotoxins) may up-regulate lymphocytes to produce cytokines that are responsible for the massive up-regulation of lymphocytes, eosinophils, and macrophages, the three most common inflammatory cells found in massive nasal polyposis. PMID- 14750607 TI - Objective and subjective evaluation of endoscopic nasal surgery outcomes. AB - BACKGROUND: Chronic rhinosinusitis (CRS) symptoms include nasal obstruction, rhinorrhea, and facial pain associated with rhinosinusitis disability. When resistance to medical treatment is associated with endonasal anomalies, endoscopic nasal surgery (ENS) can be proposed. However, objective and subjective assessment criteria regarding the evaluation of ENS outcomes remain unclear. The aims of this study were to evaluate the correlation between the inflammation in the nasal mucosa, objective recordings of nasal airway resistance (NAR), subjective evaluation of symptom intensity, and the impact of ENS on patient perceived rhinosinusitis disability. METHODS: Sixty-one consecutive patients (35 men and 26 women; mean age, 37.5 years) suffering from CRS were monitored at 4 months and 2 years after ENS. All middle turbinate mucosa were analyzed for the density of nonspecific inflammatory cells. All patients scored their own subjective rhinosinusitis symptoms and complaints of rhinosinusitis disability. An active anterior rhinomanometry was performed. RESULTS: A good correlation was observed between subjective and objective NAR (p < 0.001). We found a significant correlation between the density of inflammatory cells in the nasal mucosa, subjective nasal obstruction, and the rhinosinusitis disability score (p < 0.001). Recurrent CRS was seen only in subjects with moderate to severe inflammation of the middle turbinate mucosa sampled at the first surgical intervention. Subjective rhinosinusitis symptoms, objective NAR, and rhinosinusitis disability improved significantly after ENS. CONCLUSION: The degree of inflammation seems to be a good prognostic indicator regarding CRS recurrence. Long-term outcome after ENS for CRS showed significant improvement in subjective rhinosinusitis-specific symptoms, objective NAR, and rhinosinusitis disability. PMID- 14750608 TI - Minimally invasive treatment of the nasal inverted papilloma. AB - BACKGROUND: The purpose of this work is to evaluate our results in the treatment of the nasal inverted papillomas with an endoscopic approach using a retrospective case series. METHODS: Between 1993 and 2000 we treated 27 patients with nasal inverted papillomas. All patients underwent endoscopic nasal surgery under general anesthesia. None of the inverted papillomas extended outside of the paranasal sinuses. All tissue samples underwent polymerase chain reaction and hybridization in situ to detect genetic sequences of the human papilloma virus and Epstein Barr virus. RESULTS: The study population consisted of 16 men and 11 women with a median age of 52 years (range, 22-77 years). Ten patients (37%) had undergone a previous nasal surgery. The median follow-up was 5 years (range, 2-8 years). None of the patients presented with bilateral nasal involvement or a synchronous carcinoma. Seven patients underwent an additional surgical approach (two endoscopic approaches via a Caldwel-Luc approach, four sublabial approaches via a Caldwel-Luc approach, and one external ethmoidectomy). There were no surgical complications. Two patients (7%) had recurrent papilloma 4 and 6 years after surgery and again underwent endoscopic resection. The amplification both by polymerase chain reaction and hybridization in situ for human papilloma virus and Epstein Barr virus were negative in the specimens from all patients. CONCLUSIONS: According to the literature and our own experience, we believe that the initial surgical management of primary and recurrent inverted papillomas limited to the nasal cavity and paranasal sinuses should be endoscopic sinus surgery. PMID- 14750609 TI - Sellar reconstruction: is it necessary? AB - BACKGROUND: Sellar reconstruction is practiced routinely during the transsphenoidal approach to pituitary tumor resection. This practice exposes the patient to risks of donor site complications and may interfere with measuring postoperative tumor reduction. We propose that it is not a necessary component of transsphenoidal pituitary surgery in the absence of intraoperative cerebrospinal fluid (CSF) leak. METHODS: A retrospective chart review of 45 cases of minimally invasive pituitary surgery were reviewed. Twenty-eight cases were identified with no sellar reconstruction being performed. Age, sex, revision surgery, postoperative CSF leak, days with lumbar drain, meningitis, ophthalmoplegia, visual acuity loss, postoperative epistaxis, diabetes insipidus, development of empty sella syndrome, and length of stay were investigated in these cases. RESULTS: Twenty-three cases were the primary procedure and five cases were revision surgery. Complication rates were low and compared favorably with those reported in the literature. Five cases of transient diabetes insipidus occurred. There was one postoperative CSF leak that required 4 days with a lumbar drain. No cases of empty sella syndrome developed. There were no cases of meningitis. The average length of stay was 2.9 days. CONCLUSION: Sellar reconstruction during transsphenoidal approach to pituitary tumor resection is not required for patients without evidence of an intraoperative CSF leak. This practice exposes the patient to the risks of donor site complications without reducing the rate of postoperative complications. PMID- 14750610 TI - Low-grade fibrosarcoma of the anterior skull base: endoscopic resection and repair. AB - BACKGROUND: Fibrosarcomas of the paranasal sinuses and skull base are uncommon tumors. Traditionally, "open approach" surgery remains the mainstay for treatment of choice for these tumors. METHODS: A 49-year-old man underwent resection of a right anterior skull base fibrosarcoma using the endoscopic approach. RESULTS: Close follow-up using both endoscopic and imaging methods over a period of four years has revealed a well-healed skull base with no evidence of recurrence. CONCLUSION: Significant resistance exists at present for such a technique to deal with malignant diseases of the head and neck but results from advanced centers continue to prove that this may be a technique worth mastering and improving on. PMID- 14750611 TI - Objective testing and quality-of-life evaluation in surgical candidates with chronic rhinosinusitis. AB - BACKGROUND: Discordance has been reported between patient symptoms and objective measures of disease in chronic sinusitis, such as radiographic evaluation. The objective of this study was to evaluate the association between presurgical objective studies and presurgical quality of life (QOL). This work was designed as a cross-sectional study of 90 consecutive patients presenting to a tertiary rhinology practice for surgical management of sinonasal disease. METHODS: QOL assessment was performed using disease-specific instruments (Chronic Sinusitis Survey and Rhinosinusitis Disability Index). Computed tomography (CT) scans were scored according to the Lund-Mackay CT scoring system and endoscopy evaluation was scored by the system proposed by Lund and Kennedy. RESULTS: Ninety surgical candidates were enrolled and included in the analysis. Correlation between the QOL total scores and subscale scores was excellent (r = 0.39; p = 0.0001) as was the correlation between CT and endoscopy scores (r = 0.59; p = 0.0001). In contrast, correlation between QOL and objective measures was poor. These results were not significantly influenced by subgroup analysis by diagnosis, comorbidity, and other patient factors. CONCLUSION: Preoperative objective measures of CRS disease show little, if any, correlation with disease-specific QOL measures in surgical candidates. It is likely that CT and endoscopy are measuring a different aspect of CRS disease than the QOL measures. In addition, it is possible that preoperative QOL, either alone or in combination with CT and endoscopy, may prove important in selecting patients most likely to benefit from surgery. PMID- 14750612 TI - The percutaneous columellar strut. AB - BACKGROUND: Understanding tip support mechanisms is essential for nasal surgery, especially for preservation or augmentation of tip projection. METHODS: Support of the nasal tip often is improved by the use of struts placed between the medial crura. The percutaneous columellar strut is a technique that provides additional tip support during septoplasty and closed rhinoplasty. The strut is placed via a small vertical columellar incision. RESULTS: The advantage of easy and simple insertion outweighs the minor potential disadvantage of a small, visible vertical columellar scar. It is useful in addressing tip ptosis when trauma, surgery, or aging has reduced the contribution of the septum to tip support. CONCLUSION: A detailed description of the percutaneous columellar strut technique and results of its application are presented. PMID- 14750613 TI - Intravenous antibiotics for refractory rhinosinusitis in nonsurgical patients: preliminary findings of a prospective study. AB - BACKGROUND: Some patients with persistent chronic sinusitis have either had unsuccessful surgery or have simply refused surgery. In nonsurgical candidates, when oral antibiotics and traditional therapy are unsuccessful, home intravenous (i.v.) antibiotics present an effective alternative. METHODS: Forty-five patients were assessed before treatment using a semiquantitative scoring system and visual analog scales to measure major and minor symptoms of rhinosinusitis. Computed tomography (CT) scans were assessed using the Lund-MacKay staging system; nasal endoscopic findings were documented. Seven patients had nasal endoscopy and CT results. Medical diseases known to be associated with rhinosinusitis were documented. Endoscopic-guided culture and sensitivities were obtained. After successful insertion and radiological placement confirmation, home i.v. antibiotics were administered via a peripherally inserted central catheter line. Antibiotic choice was based on the culture and sensitivity reports. Treatment continued for 6 weeks; subsequent cultures were obtained at weeks 3, 6, and 9. Nasal endoscopy was performed, and rhinosinusitis symptoms were assessed at weeks 3, 6, and 9. Follow-up CT scans were obtained at week 12 in a subset of patients. RESULTS: Significant improvement in symptom analysis was seen in patients receiving home i.v. antibiotics when previous oral antibiotic treatment and/or surgery had failed. CONCLUSION: Home i.v. antibiotics provide an excellent alternative to surgery for patients who have either had unsuccessful surgery or who have refused surgery. PMID- 14750614 TI - A comparison of the efficacy of telithromycin versus cefuroxime axetil in the treatment of acute bacterial maxillary sinusitis. AB - BACKGROUND: Telithromycin, a new ketolide, exhibits potent activity against respiratory pathogens, including resistant strains. METHODS: Five days of telithromycin (800 mg once daily) was compared with 10 days of cefuroxime axetil (250 mg twice daily) in subjects (n = 593) with acute bacterial maxillary sinusitis (ABMS). Bacteriologic sampling was accomplished by sinus puncture or nasal endoscopy. The primary efficacy variable was clinical outcome at the posttherapy/test-of-cure evaluation in clinically evaluable patients. RESULTS: Clinical cure was achieved in 85.2% of telithromycin patients and 82.0% of cefuroxime axetil patients (difference in proportions, 3.2%; 95% confidence interval, -7.1-13.4%). Satisfactory bacteriologic response rates were comparable. Treatment-emergent adverse events for both drugs were mild or moderate. The most frequently reported treatment-emergent adverse events were nausea and diarrhea. CONCLUSION: Once-daily telithromycin for 5 days was equivalent in efficacy to twice-daily cefuroxime axetil for 10 days in patients with ABMS. Telithromycin is a suitable option for short-course therapy of ABMS. PMID- 14750615 TI - Tele-ophthalmology in India. Is it here to stay? PMID- 14750616 TI - The conundrum of lenticular oncology. A review. AB - It is a long-accepted dogma in ophthalmology that the lens is a tumour-free tissue. Yet, in the lens, there is lifelong mitotic activity in the subcapsular epithelium. Therefore, these subcapsular epithelial cells must have the potential for cellular transformation. How then can we explain the fact that no scientist has ever seen a naturally occurring primary tumour of the lens in vivo? This review discusses the early work of Mann, von Hallermann, Courtois and others who addressed the issue of tumour resistance of the lens. PMID- 14750617 TI - Jules Gonin. Pioneer of retinal detachment surgery. AB - Before the turn of the 20th century, eyes with a retinal detachment were considered doomed. Contrary to other branches of ophthalmology, such as cataract extraction, the surgical treatment of retinal detachment was still in its infancy, and the surgical success rates were less than five percent. From 1902 to 1921 Jules Gonin almost single handedly changed the landscape of retinal detachment surgery forever. He recognised that the retinal break was the cause- and not the consequence as it was largely believed at the time--of the retinal detachment, and that the treatment had at all costs to comprise the closure of the break by cauterisation. He named the procedure ignipuncture, as he cauterised the retina through the sclera with a very hot pointed instrument. Despite rigorously detailed clinical observations and increasing success rates, his discovery was not readily accepted and sometimes openly opposed by a large part of the ophthalmic establishment. It was not until 1929 that he received worldwide acclaim at the International Ophthalmological Congress in Amsterdam for his surgical technique. His legacy lives on in the eye hospital in Lausanne that bears his name, in the Gonin Medal awarded by the International Council of Ophthalmology every four years for the highest achievement in ophthalmology, and in a street named after him, the very street that he used to walk from his home to the hospital every day. PMID- 14750618 TI - Ocular manifestations of Behcet's disease in Indian patients. AB - PURPOSE: To study the prevalence, manifestations and severity of ocular involvement in Indian patients with Behcet's disease. METHOD: Prospective analysis of all patients of suspected Behcet's disease between 1997 and 2001. RESULTS: A total of 19 patients were diagnosed to have Behcet's disease. Ocular manifestations were seen in 36% patients. The commonest manifestation was conjunctival ulcer (26.3%), followed by iridocyclitis (10.5%). None of the patients had posterior segment involvement or visual loss. Response to topical corticosteroids was good. CONCLUSION: The prevalence and severity of ocular lesions in Behcet's disease is relatively low in Indian patients. Conjunctival ulcers was a common finding, which is rarely reported in the world literature. PMID- 14750619 TI - Epidemiological characteristics and laboratory diagnosis of fungal keratitis. A three-year study. AB - PURPOSE: To study the epidemiological characteristics and laboratory diagnosis of fungal keratitis seen at a tertiary eye care referral centre in South India. METHODS: A retrospective review of all culture-proven fungal keratitis seen over a 3-year period, September 1999 through August 2002. RESULTS: Fungal aetiology were confirmed in 1095 (34.4%) of 3183 corneal ulcers. The predominant fungal species isolated was Fusarium spp (471; 42.82%) followed by Aspergillus spp (286; 26%). Males (712; 65.08%) were more often affected (P<0.0001). A large proportion of the patients (732; 66.85%) were in the younger age group (21 to 50 years). A majority (879; 80.27%) came from rural areas (P<0.0001), and most patients (709; 64.75%) were farmers (P<0.0001). Ocular trauma (1009; 92.15%) was a highly significant risk factor (P<0.0001) and vegetative injuries (671; 61.28%) were identified as a significant cause for fungal keratitis (P<0.0001). 172 (15.71%) patients had concurrent diabetes mellitus. The sensitivity of 10% potassium hydroxide (KOH) wet mount preparation was higher (99.23%) than Gram-stained smear (88.73%) (P<0.0001). Incidence of fungal keratitis was higher between June and September. CONCLUSION: Agricultural activity and related ocular trauma were principal causes of mycotic keratitis. A potassium hydroxide (KOH) wet mount preparation is a simple, and sensitive, method for diagnosis PMID- 14750620 TI - Familial exudative vitreoretinopathy (FEVR). Clinical profile and management. AB - PURPOSE: To report our experience with the diagnosis and management of Familial Exudative Vitreoretinopathy (FEVR) in a predominantly older Indian population.. METHODS: This prospective interventional non-comparative case series included 38 patients of FEVR and their 23 family members. The diagnosis was established by clinical examination, fluorescein angiography and family screening. Prophylactic photocoagulation/cryotherapy or surgical treatment was done depending on the severity of the disease. RESULTS: The mean age of the patients was 23.6 years. The fundus/fluorescein angiographic findings in 116 eyes of our 61 patients (6 eyes phthisical) were as follows: forty eight (41.4%) eyes had only peripheral avascular zone, 8 (6.9%) eyes had peripheral new vessels, and 35 (30.1%) eyes had retinal detachments (RD)--10 (8.6%) exudative, 5 (4.3%) tractional and 20 (17.2%) rhegmatogenous. Prophylactic photocoagulation or cryotherapy was done in 34 eyes for retinal holes, local exudative detachments and bleeding new vessels. All the eyes retained stable vision over a mean follow-up of 16 months. Only 14 RDs were suitable for surgery: scleral buckling, vitrectomy or both. The reattachment rate was 85.7% (12 of 14) and the best-corrected visual acuity (BCVA) improved to 5/60 or better in 50% of these eyes over a 2-year follow-up. CONCLUSIONS: FEVR appears to be more common than reported. Timely diagnosis and intervention is essential in view of the lifelong progression of the disease, late exacerbations, frequent involvement of family members, and poor surgical results. A high index of suspicion, family screening and early prophylaxis are recommended to prevent avoidable blindness from this underdiagnosed disease. PMID- 14750621 TI - Five-year risk of progression of ocular hypertension to primary open angle glaucoma. A population-based study. AB - PURPOSE: To report the progression of ocular hypertension (OHT) to primary open angle glaucoma (POAG) during a 5-year follow up of a population-based sample. METHODS: Twenty-nine patients diagnosed to have OHT and 110 randomly selected normals from a population-based study in 1995 were invited for ocular examination in 2000. All patients underwent a complete ophthalmic examination; including the daytime diurnal variation of intraocular pressure (IOP) and measurement of central corneal thickness (CCT). The "corrected" IOP was used for analysis. Progression to POAG was based on typical optic disc changes with corresponding field defects on automated perimetry. RESULTS: Twenty-five of the 29 persons with OHT who could be contacted were examined. After correcting for CCT, two persons were reclassified as normal. Four of 23 (17.4%; 95% CI: 1.95-32.75) had progressed to POAG. One person amongst the 110 normals progressed to normal tension glaucoma (NTG). The relative risk of progression amongst OHT was 19.1 (95% CI: 2.2-163.4). All those who progressed had bilateral OHT. The mean and peak IOP in those who progressed was 25.4 mm Hg and 29.3 mm Hg compared to 23.9 mm Hg and 25.7 mm Hg in those who did not. Those who progressed had more than 8 mm Hg diurnal variation. The diurnal variation was less than 6 mm Hg in those who did not progress. No patient developed blindness due to glaucoma. CONCLUSION: The 5-year incidence of POAG amongst OHT in this population was 17.4% (3.5% per year). Bilateral OHT, higher peak IOP and large diurnal variation may be the risk factors for progression. PMID- 14750622 TI - Endophthalmitis caused by Acinetobacter calcoaceticus. A profile. AB - PURPOSE: To report the clinical and microbiological profile of endophthalmitis caused by Acinetobacter calcoaceticus. METHODS: A retrospective study of case series of Acinetobacter calcoaceticus endophthalmitis. Outcome measures included ability to sterilise the eye, anatomical result (clear media and attached retina) and visual recovery (visual acuity > 6/60). RESULTS: Of the 20 cases studied, 10 were cases of postoperative endophthalmitis, 3 were posttraumatic, 6 were endogenous and one was bleb-related endophthalmitis. Specific features of interest observed were relative chronicity of presentation and absence of any obvious predisposing factor in endogenous endophthalmitis cases. All cases could be sterilised except one, which needed evisceration. Cases with postoperative endophthalmitis had better anatomical outcome (7/10 with attached retina and clear media) and visual outcome (4/10 regained vision > 6/18). Higher smear positivity was seen in vitreous samples (72.2%) compared to aqueous samples (37.5%). Culture positivity was higher from the vitreous cavity compared to aqueous. The organism was sensitive to ciprofloxacin in a high percentage (88.9%) of cases. CONCLUSIONS: Visual recovery in Acinetobacter calcoaceticus endophthalmitis is modest. Ciprofloxacin is the antibiotic of choice. PMID- 14750623 TI - Comparative evaluation of Teller and Cardiff acuity tests in normals and unilateral amblyopes in under-two-year-olds. AB - PURPOSE: To compare and evaluate Teller Acuity Cards (TAC) and Cardiff Acuity Cards (CAC) to assess vision in children below the age of two. METHODS: The study evaluated TAC and CAC to assess visual acuity in 90 normal children divided into three age groups, 0-6 months (group I), 6-12 months (group II) and 12-24 months (group III). 30 cases of unilateral amblyopiogenic conditions, 10 cases each of unilateral refractive error, unilateral esotropia, and unilateral cataract, were also examined. Trained optometrists carried out binocular testing followed by monocular testing, and recorded the test time in each case. RESULTS: The mean visual acuity (in Snellen units) and standard deviation (in octaves) in the three age groups of normal children, I, II, III respectively were 6/44 +/- 0.54, 6/21 +/- 0.37 and 6/21 +/- 0.41 (binocularly by TAC) and 6/46 +/- 0.80, 6/21 +/- 0.59 and 6/14.5 +/- 0.84 (binocularly by CAC). Although the time taken for testing with CAC was less, its coefficient of variance was greater for all age groups as compared to TAC. Diminution of visual acuity could be assessed correctly by both the tests in cases of strabismus and cataract, but not in some cases of refractive error. CONCLUSION: CAC is a useful and child-friendly test. It can be used clinically but may miss some cases of visually significant refractive errors. TAC is a more dependable test to assess amblyopiogenic conditions despite the use of gratings. PMID- 14750624 TI - Posterior scleritis associated with systemic tuberculosis. AB - Infective isolated posterior scleritis is rare. We report a case of isolated posterior scleritis associated with histopathologically documented systemic tuberculosis, a hitherto unreported association. The patient responded well to a combination of oral corticosteroids with antituberculosis therapy. PMID- 14750625 TI - Intravitreal gas for submacular haemorrhage. AB - Submacular haemorrhage is an important cause for sudden visual loss needing immediate intervention. We report a case of submacular haemorrhage causing profound visual loss, which resolved after intravitreal injection of perfluoropropane (C3F8) gas followed by strict prone positioning. PMID- 14750626 TI - Posterior scleritis mimicking macular serpiginous choroiditis. AB - An unusual case of posterior scleritis mimicking macular serpiginous choroiditis is reported. PMID- 14750627 TI - Optical coherence tomographic evaluation of foveal pseudocyst in the formation of macular hole. AB - Optical coherence tomographic documented progression of foveal pseudocyst in a vitrectomised eye to full thickness macular hole is reported. Muller cell alteration and other centrifugal forces are possible pathomechanisms. PMID- 14750628 TI - Congenital glaucoma associated with 22p+ variant in a dysmorphic child. AB - A case of congenital glaucoma with developmental delay and several dysmorphic features showing 22p+ chromosomal variant is reported. PMID- 14750629 TI - Presumed ocular toxoplasmosis presenting as papillitis. AB - Unilateral papillitis is a rare manifestation of ocular toxoplasmosis. However, other causes of papillitis need to be ruled out before concluding the diagnosis. PMID- 14750630 TI - Ocular cicatricial pemphigoid: all conjunctival scarring is not trachoma. PMID- 14750631 TI - Vitamin A intervention in the Thar Desert. PMID- 14750632 TI - Spectrum and clinical profile of post-cataract surgery endophthalmitis in North India. PMID- 14750633 TI - Spectrum and clinical profile of post-cataract surgery endophthalmitis in North India. PMID- 14750634 TI - Awareness of diabetic retinopathy among diabetics. PMID- 14750635 TI - Selenite cataract and its attenuation by vitamin-E in Wistar rats. PMID- 14750636 TI - Sufficient control of heartburn in endoscopy-negative gastro-oesophageal reflux disease trials. PMID- 14750637 TI - Endoscopy-negative reflux disease: what is the value of a proton-pump inhibitor test in everyday clinical practice? AB - BACKGROUND: Our aim in this study was to evaluate a simple test with proton-pump inhibitor (PPI) for use in everyday clinical practice in diagnosing endoscopy negative reflux disease. METHODS: 68 patients with heartburn and/or acid regurgitation as their main complaint, symptoms of at least 3 months' duration prior to inclusion, all with a negative gastroscopy were included. The patients were given 60 mg of lansoprazole before breakfast for 7 days. After the test week, patients had to answer one single question on a formula. 'Did you essentially, without any doubt. have less heartburn and/or acid regurgitation during the treatment?' The only alternative answers were 'Yes' or 'No'. Patients in doubt were told to answer 'No'. 24-h pH monitoring was performed at the earliest 14 days after the test. RESULTS: 65 completed the PPI test and 52 fulfilled the pH monitoring. Defined by < 4% time oesophageal pH < 4, 34 (65%) had pathological reflux. The PPI test was positive in all patients with pathological reflux but also in 17 of 18 with normal pH-metry, giving the test a sensitivity of 97% and specificity of 6%. CONCLUSION: Application of a diagnostic PPI test in clinical practice gave a high sensitivity and unusually low specificity compared to placebo-controlled studies, indicating that a test of this nature should be used with caution in everyday practice. Most patients with endoscopy-negative GORD will be diagnosed clinically. A positive test with PPI strengthens the diagnosis but has insufficient specificity to be an objective criterion alone. pH-metry should be unnecessary for the diagnosis of ENGORD in patients with typical reflux symptoms. PMID- 14750638 TI - Natural history of gastro-oesophageal reflux disease: the neglected factor. AB - The natural history of gastro-oesophageal reflux disease (GORD) is still a matter for research. The follow-up of GORD patients under placebo and after suspension of the drugs that healed the disease, along with observations of large patient populations treated with, or without, antisecretory drugs for long periods of time, may give some information on the spontaneous evolution of the disease. To single out the outcome of each patient, predictive factors based on demographic, morphologic and pathophysiologic characteristics have been devised, but their reliability is debatable. The reason for their many discrepancies could lie in the fact that, to date, the large majority of investigators have focused their research on the characteristics of gastro-oesophageal reflux and underlying motor activity, and have overlooked the resistance of the oesophageal mucosa. As many studies have shown that the severity of GORD is not proportional to the amount of reflux, we strongly believe that knowledge of the capacity of the mucosa to resist injury and repair the damage is the key to understanding why patients with low reflux may have severe disease, while patients with high reflux may not. Research on the kind and degree of mucosal defects in patients with GORD should receive more attention and because it may help to clarify the laws that govern the GORD evolution. PMID- 14750639 TI - Prevalence of low vitamin B12 and high homocysteine in serum in an elderly male population: association with atrophic gastritis and Helicobacter pylori infection. AB - BACKGROUND: Deficiency of vitamin B12 raises the serum and tissue levels of homocysteine. Atrophic corpus gastritis results in impaired secretion of intrinsic factor and may lead to malabsorption of vitamin B12 in the intestine. We examined how common an undiagnosed vitamin B12 deficiency is among elderly men in the general population and, in particular, how often this deficiency is related to atrophic corpus gastritis. METHODS: The serum level of pepsinogen I (S PGI) was assayed in a population-based sample of 12,252 men (age 51-65 years) from two cities in Finland. In this sample, all 635 men with S-PGI < 25 microg/l formed Series A ('males with atrophic corpus gastritis'). Series C (controls- 'males without atrophic corpus gastritis)' with a non-atrophic gastric corpus was formed as a random sample of men (n = 402) with S-PGI > or = 50 microg/l. Serum levels of vitamin B12 (S-B12), folate (S-Fol), total homocysteine (S-Hcy) and Helicobacter pylori antibodies (S-HpAb) were assayed in all, or in large subsamples, of the men in Series A and C. RESULTS The men in Series A had significantly lower S-B12 and S-Fol levels than those in Series C. In Series A, 172 of 613 men tested (28%) had S-B 12 < 170 pmol/ 1, and 133 men (22%) had S-B 12 in the range 170-219 pmol/l. The corresponding prevalences were 7% (P < 0.001) and 17% (P < 0.001) in Series C, respectively. The mean S-Hcy was significantly higher in Series A in men with low S-B12 than the mean S-Hcy in Series C in men with normal S-B12. The prevalence of S-Hcy > 15 ,micromol/l was 27% in Series A and 15% in Series C (P < 0.05; chi2 = 4.63). Among subjects with S-B 12 < 220 pmol/l, 46% (104 of 226 men tested) in Series A and 16% (16 of 99) in Series C had S-Hcy > or = 15 micromol/l (P < 0.001). The mean S-Hcy was significantly (P < 0.001) higher in men with S-B12 in the range 170-219 pmol/l in Series A (mean 14.6 +/- 5.0 micromol/l) than in Series C (11.3 +/- 3.0 micromol/l). It was extrapolated that 2.5% of men in the age group 51-65 years in the present study population had a low S-B12 (< 220 pmol/l) level that associated with atrophic corpus gastritis. Of these men, 72% (128 of 179 tested) had an elevated S-HpAb level. CONCLUSIONS: Low S-B12 related to atrophic corpus gastritis is relatively common (prevalence 2.5%) among elderly males in the general population. An ongoing H. pylori infection occurs in three-fourths of these cases. PMID- 14750640 TI - IL-1 RN 2/2 genotype and simultaneous carriage of genotypes IL-1 RN 2/2 and IL 1beta-511 T/T associated with oesophagitis in Helicobacter pylori-negative patients. AB - BACKGROUND: Interleukin-1 receptor antagonist genotype 2/2 is associated with a prolonged and enhanced inflammatory response. It is suspected of being a risk factor for atrophic gastritis and gastric cancer and for some autoimmune diseases. No specific genetic risk factors for oesophagitis have been identified so far and there are no reports of IL-1 polymorphism in relation to oesophageal disease. METHODS: We studied the IL-1RN, IL-1beta-511 and IL-1beta + 3953 polymorphisms in an unselected series of 142 adult patients scheduled for gastrointestinal endoscopy because of dyspepsia. The control group consisted of university staff and students (n = 179). Helicobacter pylori status was determined by antibody testing and bacterial detection. RESULTS: Endoscopic oesophagitis was noted in 40 patients. The IL-1RN 2/2 genotype was significantly more prevalent in the patients with H. pylori-negative oesophagitis than in the control subjects (27% versus 9%; OR 3.574, CI 1.23-10.35, P = 0.034) or in the dyspeptic patients (27% versus 7%; OR 5.089. CI 1.51-17.11, P = 0.009). IL-1beta 511 T/T genotype tended to be more frequent in the H. pylori-negative patients with oesophagitis than in the control subjects (P = 0.071). The strongest association was between the simultaneous carriage of genotypes IL-1RN 2/2 and IL 1beta -511 T/T and H. pylori-negative oesophagitis. where the combined genotype was more prevalent than in the control subjects (23% versus 6%; OR 4.492, CI 1.40 14.46, P = 0.012) or the dyspeptic patients without oesophagitis (23% versus 3%: OR 9.706. CI 2.12-44.42, P = 0.003). CONCLUSIONS: The results indicate that the IL-1RN 2/2 genotype and the carriage of combined genotypes IL-1RN 2/2 + IL-1beta 511 T/T are associated with H. pylori-negative oesophagitis. This is the first report on the association between IL-1 gene polymorphism and oesophagitis. PMID- 14750641 TI - 'Serological biopsy' in first-degree relatives of patients with gastric cancer affected by Helicobacter pylori infection. AB - BACKGROUND: Relatives of patients with gastric cancer are at increased risk of developing this disease, especially if they are infected by Helicobacter pylori. Moreover, H. pylori-related atrophic gastritis and hypochlorhydria are well documented risk factors for noncardia gastric cancer. Serum pepsinogen I (sPGI) and II (sPGII) levels are low in this condition. The aim of our study was to assess by means of a 'Gastropanel' blood test, including sPGI, sPGII, gastrin-17 (G-17) and antibodies anti-H. pylori (IgG-Hp). both functional and morphological features of gastric mucosa in Hp + ve subjects with a family history of gastric cancer. MATERIALS AND METHODS: Twenty-five Hp + ve subjects consecutively referred to our department for gastrointestinal complaints, selected as first degree relatives of patients suffering from gastric cancer, were enrolled in the study and then matched for sex and age with 25 dyspeptic and Hp + ve subjects with no family history of gastric neoplasia. Blood samples were taken for determination of gastropanel in all patients; in addition, antibodies against CagA were analysed. RESULTS: No statistically significant differences were detected between the two groups as regards alcohol consumption, coffee intake and smoking habits. Mean sPGI levels in Group A (83.4 +/- 58.4 microg/L) were significantly lower than those in Group B (sPGI 159.5 +/- 80.6 microg/L; P < 0.0001) as well as sPGII (12.5 microg/L = 6.24 versus 20.6 +/- 58 microg/L; P < 0.006). No statistical difference was found between the two groups in relation to G-17 levels, IgG-Hp titres and antibodies against CagA. CONCLUSION: First-degree relatives of patients with noncardia gastric cancer affected by H. pylori infection present lower sPGI and sPGII levels, possibly due to the increased frequency of atrophic lesions in these patients. PMID- 14750642 TI - In vitro sensitivity of human gastric cancer cells (HGC-27) to Helicobacter pylori cytotoxin. AB - BACKGROUND: The VacA cytotoxin produced by Helicobacter pylori is considered an important co-factor in the pathogenesis of chronic gastritis, peptic ulcer and gastric carcinoma. The toxin remains partly bound on the bacterial surface, but a certain amount is secreted and can bind receptors on gastric epithelium. The vacuolizing activity of this toxin is related to alteration of endo-lysosomic function and pore formation into plasmatic membrane. METHODS: We investigated the 'in vitro' effect of filtrates obtained from two broth cultures of H. pylori with different genotype (vacA+ and vacA-) as verified by PCR. The effect was studied on three cell lines of epithelial origin: HeLa cells (reference strain for testing vacuolization), human transformed keratinocytes HaCaT, human gastric carcinoma cells HGC-27, and on a murine leukaemia WEHI-3B. The filtrate concentrations capable of giving vacuolization (NRU test), antiproliferative and cytotoxic effects (MTT test) were determined. The modulating effect of filtrates on drug toxicity was investigated on HeLa and HGC-27 cells by testing topoisomerase inhibitors (Ciprofloxacin and Camptothecin) and non-steroidal anti inflammatory molecules (Aspirin and Indomethacin). RESULTS: Our results confirm that vacuolizing activity is present only in VacA+ filtrate and that HaCaT and HeLa cells show a similar sensitivity, whereas gastric HGC-27 cells appear significantly resistant to VacA+ activity. Although VacA filtrate does not produce vacuolisation, it affects the cell proliferation and is cytotoxic to the four cell lines. Both the VacA+ and VacA- filtrates (at non-cytotoxic concentrations) produce a decrease in drug toxicity with the unique exception of Ciprofloxacin to gastric HGC-27 cells, which in the presence of VacA+ and VacA- produces a significant increase in toxicity. CONCLUSIONS: These data suggest that products from H. pylori (other than those that have antiproliferative and toxic activity) may modulate the sensitivity of cells to drugs 'in vitro'. If this also occurs 'in vivo', we can assume that H. pylori products interfere with drug activity on gastric tissue and also with other factors (such as cytokines) with a role in the genesis of diseases in which Helicobacters are potentially involved. PMID- 14750643 TI - Prevalence of mutations of the NOD2/CARD15 gene and relation to phenotype in Spanish patients with Crohn disease. AB - BACKGROUND: We assessed the prevalence of R702W, G908R, and L1007fs coding mutations in the NOD2/CARD15 gene and the genotype-phenotype relation in Spanish patients with Crohn disease. METHODS: A cohort of 204 unrelated patients with Crohn disease and 140 healthy controls were studied. The phenotype was established before commencement of genotyping. Genotyping of the R702W, G908R, and L1007fs gene polymorphisms of NOD2/CARD15 was performed by two independent laboratories using different techniques. In the case of discordant results, specific sequencing of DNA strands was performed. RESULTS: At least one mutation was present in 32.8% of patients compared to 10.7% in controls (OR = 4.08, 95% CI 2.21 to 7.50). In patients with Crohn disease, the frequency of R702W, G908R, and L1007fs carriers was 13.7%, 8.3%, and 14.2%, respectively. Compound heterozygotes and homozygotes occurred in 3.4% and 2.9% of patients and in none of the controls. The correlation of genotype-Vienna classification showed a significant association with ileal disease (RR = 1.61, 95% CI 1.21-2.15, P = 0.001) and an inverse association with colonic localization (RR = 0.29, 95% CI 0.11-0.80, P = 0.007). There was a significant association between G908R carriership and previous appendectomy, surgical interventions, and stricturing behavior. A gene dosage effect on phenotypic characteristics was not observed. CONCLUSIONS: In a Spanish population from Madrid, mutations of the NOD2/CARD15 gene were a marker of susceptibility to Crohn disease and were associated with ileal disease. Carriers of the G908R mutation showed a stricturing disease behavior, history of appendectomy, and surgical interventions over the course of the disease. PMID- 14750644 TI - Colorectal proliferation and apoptosis in serrated versus conventional adenoma carcinoma pathway: growth, progression and survival. AB - BACKGROUND: Hyperplastic polyp, serrated adenoma and serrated adenocarcinoma form a morphological continuum, and are likely to be biologically related. The growth rate and malignant conversion rate of serrated adenoma are postulated to be higher than those of conventional adenoma. METHODS: Immunohistochemistry for M30 and Ki67 was used to compare apoptosis and proliferation in colorectal hyperplastic polyps, serrated adenomas and serrated adenocarcinomas, and in relation to conventional adenomas and adenocarcinomas. RESULTS: There was an abrupt increase in the apoptosis in carcinomas of serrated adenoma-serrated adenocarcinoma pathway, indicating that inhibition of apoptosis is not maintained in serrated adenocarcinomas. Proliferation was significantly lower at the invasive margin than in the central part in adenocarcinomas, particularly in serrated adenocarcinomas (P < 0.005, Wilcoxon). Apoptosis was lower at the invasive margin than in the central part in serrated adenocarcinomas, but not in conventional adenocarcinomas. CONCLUSIONS: Disordered regulation of apoptosis occurs later in serrated pathway than in conventional adenoma-carcinoma pathway during malignant conversion. Serrated adenoma growth is not likely to be slower than adenoma growth, as judged by the proliferation and apoptosis rates in serrated adenomas and adenomas. In cancer, the decrease of proliferation and apoptosis in the invasive margin is likely to influence the behaviour of the tumour. PMID- 14750645 TI - Gastric cancer in young patients with no alarm symptoms: focus on delay in diagnosis, stage of neoplasm and survival. AB - BACKGROUND: The test and treat strategy for Helicobacter pylori infection has raised some concern since young gastric cancer patients may have no alarm symptoms. In this study the frequency of alarm symptoms was assessed in a series of young gastric cancer patients, as well as the impact of absence of alarm symptoms on delay in diagnosis and stage of gastric cancer at diagnosis and survival. METHODS: A retrospective study was carried out on 92 gastric cancer patients < or = 45 years of age identified from databases in four hospitals between January 1985 and December 2001. Characteristics analysed included duration and features of dyspeptic symptoms, presence of alarm symptoms, time interval from the onset of symptoms to diagnosis, pTNM stage and survival. RESULTS: Of the 92 patients, 54 (58.7%) presented uncomplicated dyspepsia and 38 (41.3%) alarm symptoms. In those with uncomplicated dyspepsia, epigastric pain was the most common complaint (64.1%) followed by vomiting (30.4%), heartburn and nausea. Weight loss was the most common alarm symptom (30.4%), followed by anorexia (10.9%), dysphagia or anaemia (7.6%). The mean delay from first symptoms to final diagnosis was 16.8 +/- 13.9 weeks in patients with alarm symptoms and 29.3 +/- 39.9 weeks in patients without alarm symptoms (P:ns). Patients without alarm symptoms showed significantly less aggressive gastric cancer compared to patients with alarm symptoms in relation to TNM stage and survival (cumulative 5 year survival rate: 76% versus 49% P: 0.01). The survival rate, at 5 years, of patients without alarm symptoms, and with a history of dyspepsia of more than 24 weeks, was higher than that in patients with early diagnosis (93.4% versus 66.5%: P: 0.05). CONCLUSIONS: A large proportion of young gastric cancer patients present without alarm symptoms. Despite the delay in diagnosis, these patients have a better outcome than those with alarm symptoms. Thus the delay in diagnosis of patients without alarm symptoms does not affect survival. PMID- 14750646 TI - Phagocytosis and LPS-stimulated production of cytokines and prostaglandin E2 is different in Kupffer cells isolated from the periportal or perivenous liver region. AB - BACKGROUND: Kupffer cells can release pro-inflammatory mediators and contribute to damage, which often appears in a zonated fashion. METHODS: To assess position associated functional differences, functions of intact Kupffer cells isolated from either the periportal or perivenous acinar region of rat liver were compared. RESULTS: Kupffer cells from the periportal region phagocytosed 2-3 times more FITC-labelled zymosan particles than corresponding perivenous cells, as determined by confocal microscopy and fluorescence assay. Periportal cells also produced more TNF-alpha and IL-1beta, but less NO and PGE2, compared to perivenous cells and the stimulation by addition of lipopolysaccharides (LPS) was moderate. In contrast, after overnight culture LPS dramatically increased TNF alpha release and significantly more so in perivenous Kupffer cells (26-fold) than in periportal cells (11-fold). CONCLUSION: Our study suggests that periportal Kupffer cells are responsible for a major part of phagocytosis by the liver. The stronger LPS response of recovered perivenous Kupffer cells suggests a dominant role of these cells in pro-inflammatory events that ultimately may contribute to development of damage in this region. PMID- 14750647 TI - Intravenous n-acetylcysteine, ascorbic acid and selenium-based anti-oxidant therapy in severe acute pancreatitis. AB - BACKGROUND: To observe outcome in a cohort of patients with severe acute pancreatitis receiving multiple anti-oxidant therapy. METHODS: An observational study was carried out in 46 consecutive patients with acute pancreatitis fulfilling current Atlanta consensus criteria for severe disease. All patients received multiple anti-oxidant therapy based on intravenous selenium, N acetylcysteine and ascorbic acid plus beta-carotene and alpha-tocopherol delivered via nasogastric tube. Principal outcomes were the effect of anti oxidant supplementation on anti-oxidant levels, morbidity and mortality in patients on anti-oxidant therapy, case-control analysis of observed survival compared to predicted survival derived from logistic organ dysfunction score (LODS), logistic regression analysis of factors influencing outcome and side effect profile of anti-oxidant therapy. RESULTS: Paired baseline and post supplementation data were available for 25 patients and revealed that anti oxidant supplementation restored vitamin C (P = 0.003) and selenium (P = 0.028) toward normal. In univariate survival analysis, patient survival to discharge was best predicted by admission APACHE-II score with relative risk of death increasing 12.6% for each unit increase (95% CI 6.0% to 19.6%). The mean LODS calculated on admission to hospital was 3.7 (standard error of the mean 4.1) giving a predicted mortality for the cohort of 21%. The observed in-hospital mortality was 43%. CONCLUSIONS: Case-control analyses do not appear to demonstrate any benefit from the multiple anti-oxidant combination of selenium, N acetylcysteine and ascorbic acid in severe acute pancreatitis. PMID- 14750648 TI - Inter-endoscopist variation in polyp and neoplasia pick-up rates in flexible sigmoidoscopy screening for colorectal cancer. AB - BACKGROUND: The Norwegian Colorectal Cancer Prevention study is an ongoing flexible sigmoidoscopy (FS) screening trial for colorectal cancer. Twenty-one thousand average-risk individuals, aged 50-64 years, living in two separate areas in Norway were randomly drawn from the Population Registry and invited to once only screening flexible sigmoidoscopy. Examinations were performed over 3 years, at 2 centres, by 8 different endoscopists, using the same type of equipment. The aim of the present study was to investigate possible differences between endoscopists in detecting individuals with polyps, adenomas and advanced lesions (adenomas with severe dysplasia and/or villous components and/or size larger than 9 mm and carcinoma) in flexible sigmoidoscopy screening. METHODS: The present trial comprises data from 8822 individuals, aged 55-64 years, who have undergone a flexible sigmoidoscopy. In the study period, all lesions detected by the different endoscopists were registered. Tissue samples were taken from all lesions detected. RESULTS: Detection rates varied significantly between endoscopists, ranging from 36.4% to 65.5% for individuals with any polyp, from 12.7% to 21.2% for any adenoma and from 2.9% to 5.0% for advanced lesions. In a multiple logistic regression model, the performing endoscopist was a strong independent predictor for detection of individuals with polyps (P < 0.001 ), adenomas (P < 0.001) and advanced lesions (P = 0.01). CONCLUSION: Detection rates for colorectal lesions vary significantly between endoscopists in colorectal cancer screening. Establishing systems for monitoring performance in screening programmes is important. Supervised training and re-certification for endoscopists with poor performance should be considered. PMID- 14750649 TI - Gastrointestinal complications of von Recklinghausen's disease: two case reports and a review of the literature. AB - There are few reports of the association between neurofibromatosis (von Recklinghausen's disease) and large, solid stromal tumours of the gastrointestinal tract. The prevalence of gastrointestinal involvement in von Recklinghausen's disease has been estimated at 11%-25%. Some associated gastrointestinal stromal tumours present clinically as bowel obstruction, perforation or gastrointestinal bleeding. We recently treated two patients with this condition who presented with gastrointestinal bleeding and were diagnosed with gastrointestinal stromal tumours. We report the unique aspects of these cases and discuss the diagnostic and management problems that are posed by this unusual association. PMID- 14750650 TI - Unilateral inner ear damage results in lasting changes in hippocampal CA1 field potentials in vitro. AB - We investigated the effects of a surgical lesion of one vestibular inner ear (unilateral vestibular damage [UVD]) on the field potential responses of CA1 neurons in vitro. Hippocampal slices were removed from rats at 4-6 weeks or 5-6 months post-UVD, and the field responses of CA1 neurons to electrical stimulation of the Schaffer collateral commissural pathway were analyzed. Compared with slices from sham and naive control animals, slices from UVD animals at 5-6 months post-UVD exhibited decreases in the population spike amplitude, the somal field excitatory postsynaptic potential (sfEPSP) slope, and the field EPSP (fEPSP) slope. For the population spike amplitude and fEPSP slope, this effect was observed in both CA1 ipsilateral and contralateral to the UVD. On both the ipsilateral and contralateral sides, paired-pulse testing showed increases in paired-pulse inhibition at the shortest interstimulus intervals (ISIs), with increases in paired-pulse facilitation at longer ISIs. This study provides the first evidence that peripheral vestibular damage can produce long-term changes in hippocampal electrophysiological activity in vitro. PMID- 14750651 TI - Retrograde amnesia and the volume of critical brain structures. AB - There are many controversies concerning the structural basis of retrograde amnesia (RA). One view is that memories are held briefly within a medial temporal store ("hippocampal complex") before being "consolidated" or reorganised within temporal neocortex and/or networks more widely distributed within the cerebral cortex. An alternative view is that the medial temporal lobes are always involved in the storage and retrieval (reactivation) of autobiographical memories (multiple trace theory). The present study used quantitative magnetic resonance imaging (MRI) in 40 patients with focal pathology/volume loss in different sites, to examine the correlates of impairment on three different measures of RA. The findings supported the view that widespread neural networks are involved in the storage and retrieval of autobiographical and other remote memories. Brain volume measures in critical structures could account for 60% of variance on autobiographical memory measures (for incidents and facts) in diencephalic patients and for 60-68% of variance in patients with frontal lesions. Significant correlations with medial temporal lobe volume were found only in the diencephalic group, in whom they were thought to reflect thalamic changes, but not in patients with herpes encephalitis or hypoxia in whom the temporal lobes were particularly implicated. The latter finding fails to support one of the main predictions of multiple trace theory, as presently expounded. PMID- 14750652 TI - Neuropsychological correlates of hippocampal and rhinal cortex volumes in patients with mesial temporal sclerosis. AB - Considerable progress has been made toward understanding the function of the primate rhinal cortex, comprising the entorhinal (ErC) and perirhinal (PrC) cortices. However, translating animal models to human memory has been limited by the technological problems associated with characterizing neural structures in vivo. Functional correlates of hippocampal and rhinal cortex volume changes were examined in a sample of 61 temporal lobe epilepsy patients with mesial temporal sclerosis (MTS; 33 left, 28 right). Patients were administered the Wechsler Adult Intelligence Scale (revised or third edition), the Wechsler Memory Scale (revised or third edition), and a spatial maze task. Neuropsychological data, together with rhinal cortex and hippocampal volumes, collected in our earlier study (O'Brien CE, Bowden SC, Whelan G, Cook MJ, unpublished observations), were analyzed using multiple regression. The only significant predictor of verbal memory function was the difference score between the volume of left hippocampus and the left PrC. Spatial maze scores were predicted by the bilateral sum of ErC volume. The difference score between the left hippocampus and left PrC volumes was the most powerful predictor of verbal episodic memory. Right hippocampal volume was not a significant predictor of nonverbal episodic memory. Verbal and nonverbal semantic memory were not significantly predicted by any combination of rhinal cortex structures. This quantitative study suggests a lateralized or material-specific memory function for the left hippocampus and left PrC, in contrast to the bilateral role of the ErC. The left hippocampus and left PrC appear to act on verbal memory function through an opposing relationship. Finally, differentiation between hippocampal and subhippocampal components in terms of episodic and semantic memory, respectively, could not be supported by the current data. PMID- 14750653 TI - Estrogen alters amyloid precursor protein as well as dendritic and cholinergic markers in a mouse model of Down syndrome. AB - Individuals with Down syndrome (DS) develop the pathological hallmarks of Alzheimer's disease at an early age, later followed by memory decline and dementia. Women with DS are twice as likely to undergo early menopause, and levels of estradiol correlate with onset of cognitive decline in these women. We have demonstrated that a mouse model of DS, mice with segmental trisomy of chromosome 16 (Ts65Dn), develop a significant deficit in both reference and working memory as young adults (6-10 months of age), coupled with phenotypic loss of cholinergic neurons in the basal forebrain and altered growth factor levels. In the present study we examined cholinergic and dendritic markers in the hippocampal formation and levels of the amyloid precursor protein (APP) in different brain regions of Ts65Dn mice treated with estradiol for 60 days. The density of the dendritic marker Map2 was significantly decreased in the hippocampal formation of middle-aged trisomic mice (9-15 months old), and the density of cholinergic neurites (acetylcholinesterase [AChE] histochemistry) was also decreased in specific layers of the hippocampus. Treatment with 17beta estradiol alleviated the decreases in Map2 and AChE staining, but had no effect on full-length APP levels in the hippocampus. In contrast, a main effect of treatment on APP levels in the striatum was noted, with significant elevations observed in controls and trisomics. These findings demonstrate that estrogen can alleviate deficits in cholinergic and dendritic elements in the hippocampal formation and further strengthens the rationale to explore estrogen replacement therapy in women with DS. PMID- 14750655 TI - Projections from the periamygdaloid cortex to the amygdaloid complex, the hippocampal formation, and the parahippocampal region: a PHA-L study in the rat. AB - The periamygdaloid cortex, an amygdaloid region that processes olfactory information, projects to the hippocampal formation and parahippocampal region. To elucidate the topographic details of these projections, pathways were anterogradely traced using Phaseolus vulgaris leukoagglutinin (PHA-L) in 14 rats. First, we investigated the intradivisional, interdivisional, and intra-amygdaloid connections of various subfields [periamygdaloid subfield (PAC), medial subfield (PACm), sulcal subfield (PACs)] of the periamygdaloid cortex. Thereafter, we focused on projections to the hippocampal formation (dentate gyrus, hippocampus proper, subiculum) and to the parahippocampal region (presubiculum, parasubiculum, entorhinal, and perirhinal and postrhinal cortices). The PACm had the heaviest intradivisional projections and it also originated light interdivisional projections to other periamygdaloid subfields. Projections from the other subfields converged in the PACs. All subfields provided substantial intra-amygdaloid projections to the medial and posterior cortical nuclei. In addition, the PAC subfield projected to the ventrolateral and medial divisions of the lateral nucleus. The heaviest periamygdalohippocampal projections originated in the PACm and PACs, which projected moderately to the temporal end of the stratum lacunosum moleculare of the CA1 subfield and to the molecular layer of the ventral subiculum. The PACm also projected moderately to the temporal CA3 subfield. The heaviest projections to the entorhinal cortex originated in the PACs and terminated in the amygdalo-entorhinal, ventral intermediate, and medial subfields. Area 35 of the perirhinal cortex was lightly innervated by the PAC subfield. Thus, these connections might allow for olfactory information entering the amygdala to become associated with signals from other sensory modalities that enter the amygdala via other nuclei. Further, the periamygdalohippocampal pathways might form one route by which the amygdala modulates memory formation and retrieval in the medial temporal lobe memory system. These pathways can also facilitate the spread of seizure activity from the amygdala to the hippocampal and parahippocampal regions in temporal lobe epilepsy. PMID- 14750654 TI - Prenatal administration of morphine decreases CREBSerine-133 phosphorylation and synaptic plasticity range mediated by glutamatergic transmission in the hippocampal CA1 area of cognitive-deficient rat offspring. AB - The central nervous system (CNS) exhibits remarkable plasticity in early life and can be altered significantly by various prenatal influences. We previously showed that prenatal exposure to morphine altered kinetic properties of N-methyl-D aspartate (NMDA) receptor-mediated synaptic currents in the hippocampus of young rat offspring at the age of 14 days (P14). The present study further investigates whether NMDA receptor-mediated synaptic plasticity and/or cyclic adenosine monophosphate-responsive element-binding protein (CREBSerine-133), an important transcription factor underlying learning and memory, can be altered by prenatal morphine exposure in these offspring. Subsequently, the Morris water maze task was performed at the older ages (P28-P31). The magnitude of long-term depression (LTD) generated by a low-frequency stimulation (LFS, 1 Hz for 15 min) in hippocampal slices from the vehicle-control offspring (P14) was significantly larger than that in slices from the morphine-treated offspring, although there was no such difference in the magnitude of long-term potentiation (LTP) elicited by a high-frequency stimulation (100 Hz for 1 s) between the two groups. Comparison of the expression range of glutamatergic synaptic plasticity in slices from the vehicle-control and morphine-treated offspring, calculated as the difference in the maximal magnitude between LTP and LTD, demonstrated a remarkably smaller range in the slices from the morphine-treated offspring. In addition, the decreased phosphorylation of CREBSerine-133 and the impaired ability of spatial learning were also seen in the morphine-treated offspring, as compared with the vehicle-control offspring. Collectively, the study suggests that maternal exposure to morphine reduces the range of synaptic plasticity by decreasing the expression of LTD, but not of LTP, in CA1 pyramidal neurons of the hippocampus from rat offspring. More importantly, decreased phosphorylation of CREBSerine-133 may play a role for the impaired spatial learning and memory in rat offspring exposure to prenatal morphine. Thus, the findings here may provide important insights into cellular/molecular mechanisms underlying pathophysiological changes in the CNS of young offspring from morphine-addicted mothers and serve as a basis for possible therapeutic intervention. PMID- 14750656 TI - Morphological and numerical analysis of synaptic interactions between neurons in deep and superficial layers of the entorhinal cortex of the rat. AB - Neurons providing connections between the deep and superficial layers of the entorhinal cortex (EC) constitute a pivotal link in the network underlying reverberation and gating of neuronal activity in the entorhinal-hippocampal system. To learn more of these deep-to-superficial neurons and their targets, we applied the tracer Neurobiotin pericellularly in layer V of the medial EC of 12 rats. Labeled axons in the superficial layers were studied with light and electron microscopy, and their synaptic organization recorded. Neurobiotin labeled layer V neurons displayed "Golgi-like" staining. Two major cell types were distinguished among these neurons: (1) pyramidal neurons with apical spiny dendrites traversing all layers and ramifying in layer I, and (2) horizontal neurons with dendrites confined to the deep layers. Labeled axons ramified profusely in layer III, superficially in layer II and deep in layer I. Analysis of labeled axon terminals in layers I-II and III showed that most synapses (95%) were asymmetrical. Of these synapses, 56% occurred with spines (presumably belonging to principal neurons) and 44% with dendritic shafts (presumably interneurons). A small fraction of the synapses (5%) was of the symmetrical type. Such synapses were mainly seen on dendritic shafts. We found in two sections a symmetrical synapse on a spine. These findings suggest that the deep to superficial projection is mainly excitatory in nature, and that these fibers subserve both excitation and feed-forward inhibition. There is an additional, much weaker, inhibitory component in this projection, which may have a disinhibitory effect on the entorhinal network in the superficial layers. PMID- 14750657 TI - Conduction latency along CA3 hippocampal axons from rat. AB - Relatively few physiological studies have been carried out on intrahippocampal axons. We have recorded compound potentials from fiber groups and the activity of individual axons at 22-25 degrees C to characterize the conduction in subsets of the broad fan-shaped CA3 pyramidal axonal tree, including the Schaffer collaterals and longitudinal branches. The same wide axonal branching was indicated by antidromic activation of individual CA3 pyramidal cells. The average compound action potential latency from the CA3 to the CA1 area (Schaffer collaterals) increased by 4.16 +/- 0.06 ms/mm separation between the stimulation and registration electrodes. The impulses spread 31% faster in the 45-degree oblique temporal than in the transverse direction across CA1. The latency of the longitudinal axons in the CA3 area increased by 6.19 +/- 0.19 ms/mm. More impressive than these direction-dependent differences in latency were the large differences between individual axons running in the same direction. For both the longitudinal axons and the Schaffer collaterals, there was a broad distribution of antidromic latencies for a given distance between the stimulation and recording points. Typically, the fastest impulses arrived in half the time of the slowest. The distribution of compound action potential latencies between two points in the tissue could be made narrower by surgical restriction of the thickness and width of the preparation. By comparison, the cerebellar parallel fibers showed a narrower distribution of their latencies than the Schaffer collaterals. Because the cerebellar fibers run more straight than Schaffer collaterals, this suggested that some of the latency differences of the latter were due to differences in the path length of the axons. One consequence of our findings is that synchronous firing of neighboring CA3 pyramidal cells does not necessarily give synchronous inputs to common target CA1 neurons. PMID- 14750658 TI - Retrograde and anterograde object recognition in rats with hippocampal lesions. AB - Retrograde and anterograde object-recognition memory was assessed in rats with cytotoxic lesions of the hippocampal formation (HPC), using a paradigm based on the natural tendency of rats to spend more time exploring novel objects than familiar objects. The rats were allowed to explore a sample object for 5 min/day on 5 consecutive days, either 5 weeks or 1 week before surgery. After surgery, retrograde recognition was assessed by comparing the amount of time spent exploring the sample versus a novel object in a free-choice situation. Control rats spent more time exploring the novel object than the sample objects from both presurgery time periods, whereas rats with HPC lesions did not discriminate between the novel objects and sample objects from either presurgery time period. Despite their deficits on the retrograde recognition test, the rats with HPC lesions performed like control rats on anterograde recognition tests, displaying a strong exploratory preference for novel objects over sample objects, with retention delays of either 15 min or 24 h. The findings suggest that extrahippocampal circuitry is capable of supporting object recognition, but only if the HPC does not participate in encoding the original encounter with the object. PMID- 14750659 TI - Differential neuronal fates in the CA1 hippocampus after hypoxia in newborn and 7 day-old rats: effects of pre-treatment with MK-801. AB - The brain displays an age-dependent sensitivity to ischemic insults. However, the consequences of oxygen deprivation per se in the developing brain remain unclear, and the role of glutamate excitotoxicity via N-methyl-D-aspartate (NMDA) receptors is controversial. To gain a better understanding of the mechanisms involved in the cerebral response to severe hypoxia, cell damage was temporally monitored in the CA1 hippocampus of rat pups transiently exposed to in vivo hypoxia (100% N2) at either 24 h or 7 days of age. Also, the influence of a pre treatment with the NMDA receptor antagonist MK-801 (5 mg/kg, i.p.) was examined. At both ages, morphometric analyses and cell counts showed hypoxia-induced significant neuronal loss (30-35%) in the pyramidal layer, with injury appearing more rapidly in rats exposed at 7 days. Morphological alterations of 4,6 diamidino-2-phenylindole (DAPI)-labeled nuclei, DNA fragmentation patterns on agarose gels, as well as expression profiles of the apoptosis-related regulatory proteins Bax and Bcl-2 showed that apoptosis was prevalent in younger animals, whereas only necrosis was detected in hippocampi of rats treated at 7 days. Moreover, pre-treatment with MK-801 was ineffective in protecting hippocampal neurons from hypoxic injury in newborn rats, but significantly reduced necrosis in older subjects. These data confirm that hypoxia alone may trigger neuronal death in vivo, and the type of cell death is strongly influenced by the degree of brain maturity. Finally, NMDA receptors are not involved in the apoptotic consequences of hypoxia in the newborn rat brain, but they were found to mediate necrosis at 7 days of age. PMID- 14750660 TI - Analysis of the presynaptic signaling mechanisms underlying the inhibition of LTP in rat dentate gyrus by the tyrosine kinase inhibitor, genistein. PMID- 14750661 TI - Small is different: energetic, structural, thermal, and mechanical properties of passivated nanocluster assemblies. AB - We explore, with the use of extensive molecular dynamics simulations, several principal issues pertaining to the energetics of formation of superlattices made through the assembly of passivated nanoclusters, the interactions that underlie the cohesion of such superlattices, and the unique mechanical, thermal and structural properties that they exhibit. Our investigations focus on assemblies made of crystalline gold nanoclusters of variable sizes, passivated by monolayers of alkylthiol molecules. An analytic optimal packing model that correlates in a unified manner several structural characteristics of three-dimensional superlattice assemblies is developed. The model successfully organizes and systematizes a large amount of experimental and simulation data, and it predicts the phase-boundary between different superlattice structural motifs that evolve as a function of the ratio between the chain-length of the extended passivating molecules and the radius of the underlying gold nanocluster. The entropic contribution to the formation free energy of the superlattice assembly is found to be large and of similar magnitude as the potential energy component of the free energy. The major contribution to the cohesive potential energy of the superlattice is shown to originate from van der Waals interactions between molecules that passivate neighboring nanoclusters. The unique mechanical, thermal, thermomechanical, and thermostructural properties of passivated nanocluster assemblies, are discussed. PMID- 14750662 TI - Shape control of III-V semiconductor nanocrystals: synthesis and properties of InAs quantum rods. AB - A novel approach for synthesis of soluble semiconductor quantum rods using metal nanoparticles to direct and catalyze one-dimensional growth is developed. The method is useful in particular for III-V semiconductors with cubic lattice, where the utilization of surfactant-controlled rod-growth is not easily realized. The growth takes place via the solution-liquid-solid (SLS) mechanism where proper precursors are injected into a coordinating solvent. Centrifugation is used for separation of rod-fractions with different lengths. The reaction is demonstrated for InAs, InP and GaAs. Focusing on InAs rods as a model system, we examined the effects of the type of metal catalyst, and the tuning of reaction conditions with respect to temperature, concentration, catalyst content and reaction time. Within the three types of metal catalysts used--Au, Ag and In, Au was found to provide the best control for achieving rod-growth even though the melting point of bulk gold is significantly higher then the reaction temperature. The structural properties of the rods were characterized by transmission electron microscopy, X ray diffraction and energy dispersive X-ray spectroscopy. Rods have a cubic lattice and grow mainly along the [111] direction. The relative gold content decreases in shorter rods suggesting Au depletion as a cause for limiting the growth. Room and low temperature absorption and photoluminescence measurements show that the band-gap shifts to the red upon increasing rod length revealing strong quantum confinement along the long axis in InAs rods, providing spectral coverage of the near-IR range relevant for telecommunication applications. Emission intensity also decreases with increased rod-length. These length dependent properties manifest the transition from 0D to 1D quantum confined systems. PMID- 14750663 TI - Photophysical and photoelectrochemical characteristics of multilayers of CdS nanoclusters. AB - Luminescence lifetime measurements in sub-ns time resolution and visible pump-IR absorption probe measurements to study photo-induced carrier dynamics using fs laser pulses were carried out for the CdS nanoclusters prepared by reverse micelle method, the nanoclusters modified with 2-mercaptoethanesulfonate (SO3 CdS) in water, and the SO3-CdS nanoclusters in a multilayer assembly. For a comparison, similar measurements were also carried out for a CdS nanocluster doped glass. The photoelectrochemical characteristics of the multilayer are also measured. The characteristics of nanocluster dispersion and those of multilayer assembly are compared. The sample dependent decay behaviors of luminescence and IR absorption are somewhat different and the importance of visible pump-IR absorption probe measurement is demonstrated. PMID- 14750664 TI - Electron-conducting quantum-dot solids with ionic charge compensation. AB - Electron-conducting quantum-dot solids can be prepared on the basis of assemblies of colloidal insulating nanocrystals if electrons can be injected in the delocalized conduction orbitals. We discuss the energetics of electron injection in such an artificial solid consisting of weakly coupled quantum dots. We show that quantum confinement and electron electron repulsion determine the charging characteristics. The electron electron repulsion energy can be screened by three dimensional charge compensation from trapped holes or positive inert ions inserted in the assembly. We present experimental results on the electron storage and long-range transport in assemblies of ZnO nanocrystals in which the electron charge is compensated by positive ions. The electron electron repulsion energy in assemblies permeated with an aqueous electrolyte solution is strongly screened. In contrast, the repulsion energy is about 100 meV in aprotic solvents; the repulsion energy strongly influences electron storage and the characteristics of long-range electron transport. PMID- 14750665 TI - Growth, conductivity, and vapor response properties of metal ion-carboxylate linked nanoparticle films. AB - Nanoparticles of metals (Au, Ag, Pd, alloys) in the size range 1-3 nm diameter can be stabilized against aggregation of the metal particles by coating the metal surface with a dense monolayer of ligands (thiolates). The stabilization makes it possible to analytically define the nanoparticle composition (for example, Au140(hexanethiolate)53, I) and to elaborate the chemical functionality of the protecting monolayer (for example, Au140(C6)35(MUA)18, II, where C6 = hexanethiolate and MUA = mercaptoundecanoic acid). Network polymer films (IIfilm) on interdigitated array electrodes can be prepared from II, based on cation coordination (i.e., Cu2+, Zn2+, Ag+, methyl viologen) by the carboxylates of MUA. The electronic conductivity of the IIfilm network polymer films occurs by electron hopping between the Au140 nanoparticle cores, and offers an avenue for investigation of metal-to-metal nanoparticle electron transfer chemistry. The report begins with a brief summary of what is known about metal nanoparticle electron transfer chemistry. The investigation goes on to assess factors that influence the dynamics of film formation as well as film conductivity, in the interest of better understanding the parameters affecting electron hopping rates in IIfilm network polymer films. Finally, sorption of organic vapors into IIfilm causes a decreased electronic conductivity and increased mass that can be assessed using quartz crystal microbalance measurements. The change in electronic conductivity can be exploited for the sensing of organic vapors. PMID- 14750666 TI - Gold-nanoparticle/organic linker films: self-assembly, electronic and structural characterisation, composition and vapour sensitivity. AB - Gold-nanoparticle/organic films were prepared via layer-by-layer self-assembly using dodecylamine-stabilised Au-nanoparticles and poly(propyleneimine) (PPI) dendrimers of generation one to five (G1-G5) or hexadecanedithiol (HDT) as linker compounds. TEM and FE-SEM images revealed that the bulk of the films consisted of nanoparticles with diameters of about 4 nm. XPS was used to study the chemical composition of the films. The C 1s and N 1s signals of an AuPPI-G4 film were interpreted qualitatively according to the dendrimer structure. The absence of the nitrogen signal in case of an AuHDT film indicated that the dodecylamine ligands were quantitatively exchanged during film assembly. About 76% of the sulfur atoms were bound to the nanoparticles. the remainder being present as free thiol (S H) groups. All films displayed linear current voltage characteristics and Arrhenius-type activation of charge transport. The conductivities of the AuPPI films decreased exponentially over approximately two orders of magnitude (6.8 x 10(-2) to 1.0 x 10(-3) ohms(-1) cm(-1)) with a five-fold increase of the dendrimer generation number. Dosing the films with solvent vapours caused their resistances to increase. Using different solvent vapours demonstrated that the sensitivity of this response was determined by the solubility properties of the linker compounds. Microgravimetric measurements showed that absorption of analyte was consistent with a Langmuir adsorption model. These measurements also revealed a linear correlation between the electrical response (deltaR/Rini) and the concentration of absorbed analyte. The absorption of d4-methanol from a saturated vapour atmosphere was studied by neutron reflectometry with an AuPPI-G4 film. This measurement indicated condensation of methanol on top of the film and a uniform distribution of the analyte across the film thickness. PMID- 14750667 TI - Optical properties of nanostructured metal films. AB - Nanostructured metal films of platinum, gold and silver up to 675 nm thick we prepared by electrochemical deposition through templates of 700 nm diameter polystyrene spheres assembled as hexagonal close packed monolayer on an evaporated gold surface followed by removal of the template by dissolution in tetrahydrofuran. The reflection spectra of the films at normal incidence were recorded as a function of film thickness and the spectra correlated with the local visual appearance of the film and the surface structure from SEM. For thin films, below one quarter sphere height, the spectra show a single reflectivity dip at a wavelength just below the sphere diameter consistent with surface plasmon grating-like behaviour. For the thicker films several reflectivity dips are observed which move towards longer wavelength with increasing film thickness. This behaviour is shown to be consistent with a model in which light reflected from the top of the structure interferes with light reflected from within the spherical segment cavities in the film. PMID- 14750668 TI - AuAg bimetallic nanoparticles: formation, silica-coating and selective etching. AB - A time-resolved study of the formation of AuAg alloy nanoparticles during boiling of AgNO3 and HAuCl4 in the presence of sodium citrate has been performed by monitoring the UV-visible spectra of the solutions. This study reveals clear differences with respect to the formation of pure Au particles, and suggests that gold and silver nanoparticles nucleate separately, but lattice rearrangement eventually leads to formation of alloy nanoparticles with the expected composition, which was confirmed by high resolution TEM. Additionally, we studied the chemical reaction of the alloy nanoparticles with NH4OH, observing that part of the Ag atoms get oxidized, but reorganization of the crystal lattice during the reaction prevents full transformation into pure gold nanoparticles. This allows the synthesis of silica-coated bimetallic particles with tailored plasmon resonance between ca. 430 and 520 nm, as well as homogeneous incorporation of the coated alloy nanoparticles within silica gels. PMID- 14750669 TI - Hyper-Rayleigh scattering of gold nanorods and their relationship with linear assemblies of gold nanospheres. AB - The surface plasmon enhanced hyper-Rayleigh scattering light collected from an aqueous solution of gold nanorods is reported. A non negligible part of the signal is attributed to a photoluminescence background attributed to the electron hole recombination following multiphoton excitation of d-valence band electrons into the sp-conduction band. This radiative relaxation process is likely favored by the presence of the organic species adsorbed at the surface of the nanorods. The absolute value for the hyperpolarisability of nanorods is also compared by the external reference method to that of para-nitroaniline and found to be rather large although an absolute value cannot be given because the exact number density of the gold nanorods is unaccessible. This value is however compared with values reported for linear assemblies of gold spherical nanoparticles and further support the simple model of gold metal ellipsoids to describe the hyper-Rayleigh light intensities. The polarisation analysis of the hyper-Rayleigh scattering light is also determined for gold nanorods and compared to the expected one for gold nanospheres. For the latter spheres, the weakness of the signal intensities precludes a definite comparison with the model. On the opposite, for the nanorods, the polarisation dependence of the hyper-Rayleigh scattered light clearly deviates from the one expected for nanospheres. PMID- 14750670 TI - Electrochemical and optical properties of two dimensional electrostatic assembly of Au nanocrystals. AB - The spectroscopic and electrochemical properties of two-dimensional electrostatic assembly of Au nanocrystals are examined on poly-L-lysine (pLys) modified gold electrodes. The surface preparation for the nanoparticle deposition involved the self-assembly of a monolayer of 11-mercaptoundecanoic acid on the electrode surface, followed by the electrostatic deposition of pLys from aqueous solution. The polyelectrolyte layer acts as the electrostatic anchor for the Au particles. Electrostatically stabilised Au particles were prepared by homogeneous reduction in the presence of citrate, yielding monodispersed colloidal suspension with an average diameter of 18 +/- 2 nm. After 4 h of deposition, the citrate-stabilised particles reach a maximum surface density of (8.2 +/- 0.1) x 10(10) particles cm( 2), with an average edge-to-edge distance of 25 nm. The particle surface density was estimated from scanning electron micrographs. Kelvin probe measurements were employed for examining changes in surface dipole introduced by the 2D array of nanocrystals. From simple electrostatic arguments, the apparent static dipole moment per particle was estimated of the order of 2700 D. The strong interaction between the nanocrystals and the pLys layer is responsible for the surface charge displacement, leading to changes in the surface dipole of 0.35 eV. These electrostatic interactions also manifest itself by the red shift of the plasmon resonance of the assembly with respect to the aqueous colloidal suspension. Analysis of the spectral broadening was attempted within the framework of the so called coherent-potential approximation. Finally, electrochemical studies in 1,2 dichloroethane show a large electronic overlap between the nanocrystals and the metal substrate. Results obtained from electrochemical impedance spectroscopy strongly suggest that the electrostatic assembly of nanocrystal behaves like a 2D array of randomly distributed spherical nanoelectrodes. PMID- 14750671 TI - Electron exchange between two electrodes mediated by two electroactive adsorbates. AB - Experimental data for electron exchange between two electrodes covered by electroactive films are presented and discussed in terms of the Gerischer model. A model Hamiltonian is proposed for such indirect electron exchange involving two intermediate species. Explicit model calculations are performed for the case in which the coupling between the two adsorbates is weak and determines the overall rate. The calculations agree well with the experimental data, and can be used to determine the energy of reorganization associated with the electron transfer. PMID- 14750672 TI - Wiring nanoparticles with redox molecules. AB - Gold nanoparticles were used to make electrical contact to redox-active organic molecules. Viologen based dithiols were self-assembled from solution on Au(111) for use as tethers to attach nanoparticles to a conducting substrate. The topography and electrical properties of the resulting films were investigated by STM and STS and the orientation of these linkers was investigated by FTIR. Surface coverage increased with increasing reaction time, resulting in a change of film orientation from a flat to a more upright standing conformation. Gold nanoparticles attached to these self-assembled films were characterised by STM. It was possible to isolate a single redox-active molecule in an alkanethiol matrix and by subsequent attachment of a single gold nanoparticle the electrical properties of single wired molecules could be investigated. This method allowed the measurement of the conductivity of single molecules connecting a nanoparticle to the substrate chemically, thus forming stable electrical contacts at both ends. PMID- 14750673 TI - Adaptive chemistry of bifunctional gold nanoparticles at the air/water interface. A synchrotron X-ray study of giant amphiphiles. AB - A series of ligand stabilized gold nanoparticles with diameters close to 3 nm were studied as Langmuir monolayers at the air/water interface by synchrotron X ray diffraction and reflectivity. Alkylthiols with different length and/or terminal functional group (hydrophilic or hydrophobic) were introduced into the ligand shell by ligand place exchange reactions. Synchrotron grazing incidence X ray diffraction (GIXD) and specular X-ray reflectivity reveal the well known hexagonally packed monolayers. In addition the mixed hydrophilic/hydrophobic ligand shell nanoparticles show a high degree of environmental responsiveness, as they adapt to an amphiphilic distribution of ligands around the gold core when spread at the water surface. Likewise nanoparticles of mixed long and short alkyl chains respond to lateral pressure by adopting a structure where the short alkyl chains determine the in-plane nearest neighbor distance while the long alkyl chains determine the film thickness. Based on X-ray reflectivity measurements, which quantitatively account for the electron density in the monolayers, combined with GIXD we calculate the average size and number of atoms of the individual gold particle cores, and estimate the number of passivating ligand on the particle surface. Typical values are dAU-core = 16 A, dnanoparticle = 26-37 A depending on the ligands, Mw = 30-40000 g mol(-1) number of ligands = 40-60. The thickness of the monolayers was determined by AFM after transfer of the monolayers to a solid support using the Langmuir Schaefer technique. The combination of the different techniques produce a very consistent picture of the structure and adaptive chemical nature of the nanoparticles studied, and reveal a surprisingly monodisperse particle distribution centered around 140 atoms in the gold core. PMID- 14750674 TI - Immobilisation and synthesis of DNA on Si(111), nanocrystalline porous silicon and silicon nanoparticles. AB - Oligonucleotides have been synthesized on hydrogen-terminated Si(111) and porous silicon using surface hydrosilation of difunctional molecules (1,(omega) dimethoxytritylundecenol) to produce a monolayer bearing suitable reactive groups to allow automated solid-phase DNA synthesis. The absence of an intervening oxide enables electrochemical characterisation of the surface-bound oligonucleotides. Complementary sequences to the DNA synthesized on Si(111) undergo hybridisation at the surface and a straightforward electrochemical quantitation of the amount of synthesized DNA and its hybridisation efficiency (47%) is possible using Ru(NH3)6(3+) as a redox label. In the case of DNA synthesized in porous silicon, electron transfer (ET) between DNA and the underlying bulk semiconductor can be studied by cyclic voltammetry, however the anisotropic diffusion inside the porous layer and the large resistance of the porous silicon results in voltammograms for which thin-layer behaviour is not observed and the peak currents increase with the square root of scan rate. We interpret these voltammograms in terms of charge transport limitations in the layer of metal centres bound to the DNA inside the pores. Further evidence for this interpretation has been obtained using scanning electrochemical microscopy (SECM) to study the charge transport between redox species in films of DNA synthesized on Si(111) surfaces that are in contact with an aqueous phase. As the bulk concentration of Ru(NH3)6(3+) is reduced below about 250 microM the SECM feedback indicates that the rate of charge transport between surface-bound Ru(NH3)6(3+) exceeds that due to diffusion in the liquid phase. Electrochemical quantitation of the DNA is not possible in this situation, however we have been able to obtain independent determinations using radioassay based on 32P or UV/VIS spectrophotometry of dimethoxytrityl cation cleaved from the porous layer. In the case of the former, use of labelled complementary sequences shows an inverse relationship between the current density used to prepare the porous silicon and the amount of hybridisation. This can be interpreted in terms of the specific surface area of the porous silicon layers since the hybridisation efficiencies (ca. 40%) obtained by comparing DMT+ cleaved from sequences synthesized on the surface and then from complementary sequences after hybridisation were relatively insensitive to the current density used to prepare the layers. Our recent work has also been concerned with individual Si nanocrystals generated by breaking up porous silicon during thermal hydrosilation reactions. FTIR spectroscopy shows these particles are also coated with an organic Si-C-bonded monolayer and they form stable, non-turbid and strongly luminescent (lambdamax = 600-650 nm) dispersions in apolar solvents (L. H. Lie, M. S. Duerdin, E. M. Tuite, A. Houlton and B. R. Horrocks, J. Electroanal. Chem., 2002, 538/539, 183). The effect of carrying out synthetic reactions on the porous silicon prior to breaking up the layer is to produce instead larger, micron-scale assemblies with a nanometre scale internal structure. Micron-sized particles of porous silicon produced by breaking up the layer can be probed by confocal Raman spectroscopy using the electric field of a focused laser to trap such particles. Although these particles are also luminescent, the use of relatively long wavelength laser excitation (lambda = 785 nm) allows acquisition of Raman spectra from individual particles in the optical trap. The bulk optical phonon mode at ca. 520 cm(-1) characteristic of crystalline silicon is red-shifted and broadened providing evidence for an internal nanometre scale substructure in these micron-sized particles and we also see evidence for this mode in the colloidal suspensions of the Si nanoparticles. We propose a model for the formation of these two types of particles and briefly discuss the prospects to extend our solid-phase synthesis on porous silicon to allow the facile synthesis of luminescent Si nanocrystals bearing DNA or other biomolecules. PMID- 14750675 TI - Self assemblies of nanocrystals: preparation, collective properties and uses. AB - Self-organization of nanocrystals depends on the type of nanomaterial and the coating. With silver nanocrystals, the self-organization is partially perturbed by the latter. With cobalt nanocrystals, the size distribution and thus the self organization is controlled by the amount of reducing agent added during the chemical reaction and not by the micellar solution. It is possible to make "supra" crystals of cobalt nanoparticles in a face centered cubic (fcc) structure. By applying, during the evaporation process, an external magnetic field to ferrite nanocrystals dispersed in solution, nanocrystal organizations markedly change with their coating. Hence tubes are obtained with nanocrystals coated with citrate ions whereas thick films are produced when the coating is replaced by dodecanoic acid. Collective magnetic properties due to the organization in tubes are observed with a behavior similar to that observed with nanowires. The use of nanocrystals as a mask to produce various patterns on silicon is described. PMID- 14750676 TI - Magnetic nanoparticles through organometallic synthesis: evolution of the magnetic properties from isolated nanoparticles to organised nanostructures. AB - Co and NiFe nanoparticles (2.7 to 3.3 nm mean diameter) of narrow size distribution have been obtained through the decomposition of organometallic precursors in organic solutions of long alkyl chain ligands, namely oleic acid and hexadecylamine. Materials of various volume fractions were produced. The particles have been structurally characterised by WAXS. Both adopt the bulk structure: HCP in the case of cobalt; a mixture of FCC and BCC for NiFe. Their aptitude to self-assemble either on flat supports or in bulk solid state has been investigated by means of TEM and SAXS. This study suggests the crystallisation of the nanoparticles upon solvent evaporation, especially a local FCC arrangement was observed for the NiFe material. Magnetic measurements (SQUID) confirm this tendency. The blocking temperature depends on the metal volume fraction, i.e. on the anisotropy generated by the dipolar couplings (Ki). We show that, for dense samples, the particles of high intrinsic anisotropy, Ku, (Co) still display an individual behaviour while the soft ones (NiFe) display a collective behaviour. PMID- 14750677 TI - Spin-labelled Au nanoparticles. AB - A series of Au nanoparticles functionalised with nitroxide spin labels has been prepared and studied by EPR spectroscopy. Samples with low coverage of the spin label were used to investigate the dynamics of the surface-attached labels at different distances from the Au surface. The rotational correlation times of spin labels vary from 10(-10) s to more than 3 x 10(-9) s, depending on the chain length of the label and the surrounding ligand. The samples with higher coverage of the spin label show an increasing contribution of the exchange interaction between nitroxides adsorbed in a close proximity to each other on the same nanoparticle. Quantitative analysis of the EPR spectra of these samples suggests the presence of non-equivalent binding sites on the surface of Au nanoparticles. Additionally, EPR signals of isolated radical pairs were observed at intermediate coverage. PMID- 14750678 TI - The synthesis of matrices of embedded semiconducting nanowires. AB - In this work we report how single crystal nanowires can be assembled into regular arrays using mesoporous thin films to define the architecture. Mesoporous thin films were prepared by a sol-gel method. These provide films of very regular structure and dimensions. The films produced in this way have almost single crystal like structures and can also exhibit strong epitaxy to the underlying silicon substrate. The films are subjected to a supercritical fluid (SCF) environment in which a precursor is decomposed to yield nanowires of metals, semiconductors or oxides. Using these SCF conditions, pore filling is complete and the products are nanowires which are single crystals and structurally aligned in one direction. The growth mechanism of the nanowires is described and size effects discussed. PMID- 14750679 TI - Structure of assemblies of metal nanowires in mesoporous alumina membranes studied by EXAFS, XANES, X-ray diffraction and SAXS. AB - Mesoporous alumina membranes ("anodic aluminium oxide", or "AAO") are made by anodic oxidation of aluminium metal. These membranes contain hexagonal arrays of parallel non-intersecting cylindrical pores perpendicular to the membrane surface. By varying the anodisation voltage, the pore diameters are controllable within the range 5-250 nm. We have used AAO membranes as templates for the electrochemical deposition of metals within the pores to produce nanowires. These represent assemblies of one-dimensional quantum wires with prospective applications in electronic, optoelectronic and magnetic devices. Detailed characterisation of the structures of these nanowire assemblies on a variety of length scales is essential to understand their physical properties and evaluate their possible applications. We have used EXAFS, XANES, WAXS, high energy X-ray diffraction and SAXS to study their structure and bonding. In this paper we report the results of our studies of four different nanowire systems supported in AAO membranes. These are the ferromagnetic metals iron and cobalt, the superconducting metal tin, and the semiconductor gallium nitride. Iron nanowires in pores of diameter over the range 12 nm-72 nm are structurally very similar to bcc bulk iron. They have a strong preferred orientation within the alumina pores. Their XANES shows significant differences from that of bulk iron, showing that the electronic structure of the iron nanowires depends systematically on their diameter. Cobalt nanowires are composed of a mixture of hcp and fcc phases, but the ratio of the two phases does not depend in a simple way on the pore diameter or preparation conditions. In bulk cobalt, the fcc beta-phase is normally stable only at high temperatures. Strong preferred orientation of the c-axis in the pores was found. Tin nanowires in alumina membranes with pores diameters between 12 nm and 72 nm have a tetragonal beta-structure at ambient temperature and also at 80 K. Magnetic susceptibility measurements show that they are diamagnetic, and become superconducting at the same temperature as bulk tin (3.7 K). Gallium nitride nanowires have been prepared in alumina membranes with pore diameter 24 nm by a novel method. Gallium nitrate was deposited in the pores from aqueous solution and thermolysed at 1000 degrees C to form Ga2O3, which was reacted with ammonia at 1000 degrees C. The GaN nanowires have the wurtzite structure. Preparation at 1150 degrees C led to the incorporation of aluminium in the GaN. The mesoscopic ordering of the pores in the AAO membranes and their filling by metal nanowires has been studied by SAXS, which shows patterns of Bragg peaks arising from the pore arrays. Additionally, the cobalt nanowires have been the subject of an initial ASAXS study. PMID- 14750680 TI - Thin alumina films on Ni3Al(111): a template for nanostructured Pd cluster growth. AB - The growth of vapor deposited palladium on a well-ordered thin alumina film grown on a Ni3Al(111) surface was studied as a function of the sample temperature during deposition and the palladium flux. The superstructure of the oxide film turns out to be an excellent template for the growth of nanostructured palladium cluster arrays. By taking advantage of the growth steering properties of the alumina film we were able to prepare nearly perfectly ordered hexagonal arrays of palladium clusters with a uniform distance of 4.5 nm between the particles. Furthermore the dependence of the cluster height and diameter on the sample voltage as measured by STM has been investigated. In the range between 0.7 V and 2.0 V the measured cluster heights are nearly independent of the sample voltage whereas at voltages higher than 2 V they are continuously decreasing. PMID- 14750681 TI - Size effects on reactivity of Pt nanoparticles in CO monolayer oxidation: the role of surface mobility. AB - In the present paper we study the reactivity of model Pt nanoparticles supported on glassy carbon. The particle size effect is rationalized for CO monolayer oxidation exploring electrochemical methods (stripping voltammetry and chronoamperometry) and modelling. Significant size effects are observed in the particle size interval from ca. 1 to 4 nm, including the positive shift of the CO stripping peak with decreasing particle size and a pronounced asymmetry of the current transients at constant potential. The latter go through a maximum at low COads conversion and exhibit tailing, which is the longer the smaller the particle size. Neither mean field nor nucleation & growth models give a coherent explanation of these experimental findings. We, therefore, suggest a basic model employing the active site concept. With a number of reasonable simplifications a full analytical solution is obtained, which allows a straightforward comparison of the theory with the experimental data. A good correspondence between experiment and theory is demonstrated. The model suggests restricted COads mobility at Pt nanoparticles below ca. 2 nm size, with the diffusion coefficient strongly dependent on the particle size, and indicates a transition towards fast diffusion when the particle size exceeds ca. 3 nm. Estimates of relevant kinetic parameters, including diffusion coefficient, reaction constant etc. are obtained and compared to the literature data for extended Pt surfaces. PMID- 14750682 TI - The structure and reactivity of anchored nanoparticles on a reducible support. AB - We have used scanning tunnelling microscopy (STM) to study the structure and reactivity of metal nanoparticles on a TiO2(110) surface. We have deposited two metals (Pt and Ru) using different techniques, metal vapour deposition (MVD) for Pt and metal organic chemical vapour deposition (MOCVD) for Ru. Pt was deposited onto a (1 x 3) reconstructed surface and is found to sinter at temperatures above 873 K, where the onset of nanoparticle mobility is expected. The effect that Pt has on the rate of re-oxidation of a more reduced titania sample with a (1 x 1) terminated surface was studied. At 673 K no preferential re-growth around the particles is observed and this has been attributed to the desorption characteristics of oxygen from Pt surfaces. The results are compared to those obtained from Pd covered TiO2 where such preferential growth has previously been observed. Ru was deposited using a carbonyl precursor, Ru3(CO)12, and is found to adsorb preferentially on the (1 x 2) rows of a surface consisting of both (1 x 1) and (1 x 2) added row structures. Upon annealing the compound is initially de carbonylated to leave Ru nanoparticles on the surface, which can then sinter with increasing temperature. These particles have been alloyed with Pt and their re oxidation characteristics studied. The alloy particles alter the oxygen desorption characteristics from that of the constituent elements (Pt and Ru) and consequently an enhancement in the rate of re-oxidation of a TiO2(110) surface at 673 K is observed. PMID- 14750683 TI - Concluding remarks: nanoparticle assemblies: state-of-the-art and future challenges. PMID- 14750684 TI - Childhood leukaemia and radiation exposure of fathers--the end of the road, perhaps? PMID- 14750685 TI - The evolution of the international system of radiological protection: stakeholder views from the 1st and 2nd NEA/ICRP fora. AB - The Nuclear Energy Agency's (NEA's) Committee on Radiological Protection and Public Health (CRPPH) has collaborated closely with the ICRP in its efforts to develop new recommendations for radiological protection at the start of the 21st century. As part of this effort, the NEA organised, in collaboration with the ICRP, two fora to discuss the radiological protection of the environment (Taormina, February 2002) and the future policy for radiological protection (Lanzarote, April 2003). Both these meetings were attended by a broad representation of stakeholders. The CRPPH and other stakeholders universally appreciated the opportunity to speak directly with the ICRP on these important subjects. This report summarises the main conclusions made during these two meetings to advance the deliberations of the ICRP to create a new set of recommendations responsive to stakeholder needs, firmly rooted in science, and that can be implemented in a timely, efficient and cost-effective manner. PMID- 14750686 TI - The children of atomic bomb survivors: a synopsis. AB - When the atomic bombing of Hiroshima and Nagasaki occurred in the summer of 1945, most members of the public presumed that many of the children conceived by the survivors would be grossly deformed or seriously damaged in other ways as a consequence of radiation-induced mutations. Although the experimental data then available, largely limited to studies of Drosophila melanogaster, the common fruit fly, did not support this perception, the limitations of the data and the depth of public concern warranted a careful follow-up of the children born to the survivors. To this end a surveillance was begun in 1947 of all pregnancy outcomes after 20 weeks of gestation in these two cities. Over the half century subsequent to the initiation of this surveillance, some 80-odd thousand pregnancy outcomes have been studied and a variety of potential indicators of mutational damage measured. This report summarises the findings of these studies and offers an estimate of the genetic risk based on these findings. PMID- 14750687 TI - Comparison of Health and Safety Executive and Cumbrian birth cohort studies of risk of leukaemia/non-Hodgkin's lymphoma in relation to paternal preconceptional irradiation. AB - In 1993, a case-control study by the Health and Safety Executive (HSE) assessed the risk of leukaemia and non-Hodgkin's lymphoma (LNHL) among children of fathers employed at the Sellafield nuclear installation in relation to paternal preconceptional irradiation (PPI). It concluded that the statistical association between risk of LNHL and PPI was confined to children born in the village of Seascale, where the dose-response was extremely high and very significant. In contrast, in 2002, a Cumbrian birth cohort study, investigating largely the same cases, concluded that this statistical association was not significantly different among children born inside and outside Seascale and estimated the dose response inside Seascale to be much lower. This review makes a detailed comparison of the two studies, considering their design, data and analyses. The differences between their findings are due to: (i) differences in the distribution of offspring-years which are differential with respect to dose category and Seascale birth status, (ii) a non-Seascale high-dose case included in the Cumbrian but not the HSE study, (iii) differences between analyses using categorical and continuous PPI dose and (iv) the presence of Seascale controls with PPI over 200 mSv in the Cumbrian but not the HSE study. PMID- 14750688 TI - Application of ALARP to extremity doses for hospital workers. AB - The implementation of ALARP for hospital workers is considered in relation to extremity doses. Criteria are proposed which could provide guidance in determining strategies for both implementing radiation protection measures and dose monitoring for the extremities. Two groups of hospital workers have been studied, namely interventional radiologists/cardiologists, and radionuclide staff preparing and administering radiopharmaceuticals. The radiology procedures can give high doses to both the hands and legs. Those to the legs can be reduced by the use of lead rubber shields. Study of the distribution of dose across radiologists' hands has identified the ring position on the little finger as the appropriate position for dose monitoring. The variations in dose across the hands of radionuclide workers are greater, with the tip likely to receive the highest dose. The protection strategy will need to be determined for each department, because of the wide range in techniques used in handling radiopharmaceuticals. It is hoped that the criteria could aid balanced decision-making about the appropriate protection strategy and ensure that protection measures are in place where they are required, but avoid their introduction where they are unnecessary. PMID- 14750689 TI - Persistence of translocation frequencies in blood lymphocytes following radiotherapy: implications for retrospective radiation biodosimetry. AB - Chromosome aberration analysis using a G-banding technique was performed on peripheral blood lymphocyte cultures from eight individuals over a 5 year period following therapeutic radiation exposure. Samples were placed in three time periods comprising 0-12, 12-36 and 36-60 months post-treatment. The group was heterogeneous with respect to exposure and this resulted in wide differences in initial total translocation yields. Total translocation frequencies declined in seven of the eight cases, reaching significance in four cases. This decline was attributed to a decrease in cells, which in addition to translocations, also contained aberrations such as dicentrics which resulted in them being unstable. In all eight cases, when only stable cells were considered, no significant differences were observed in translocation frequencies between the different time periods post-treatment. Thus, although the frequency of translocations in stable cells is persistent over time, extrapolating to total initial yield, and using this to equate to dose, is not possible in cases where the exposure has been high and non-homogeneous. In practice, retrospective biological dosimetry is more often required in cases of historical, usually protracted, exposures which will have been essentially uniform and not of a sufficiently high dose for many cells to have acquired more than one aberration. In such cases the frequency of translocations observed some years after the exposure can be assumed to reflect induced frequencies and be used for dose estimation. PMID- 14750690 TI - Outdoor and indoor dose assessment using environmental thermoluminescence dosimeters (TLDs) in Costa Rica. AB - Costa Rica lies at the intersection of the Cocos and Caribbean plates in Central America. It has mountain ranges with many active volcanoes along its territory. Its soils are predominantly of volcanic origin. Natural radiation measurements utilising environmental CaF2:Dy thermoluminescence dosimeters were used for the first time in Costa Rica by the Dosimetry Section of the Atomic, Nuclear and Molecular Sciences Research Center of the University of Costa Rica. Seven hundred outdoor measurements were obtained in a 3.5-year period at eight different sites throughout the country. One hundred and seventy-four indoor readings were also collected at four sites for a 2-year period. Population-weighted averages give 82 nGy h(-1) for outdoors and 130 nGy h(-1) for indoors. The values lie on the upper range of worldwide reported values due to reported soil characteristics rich in uranium and potassium. A preliminary population-weighted value of 0.74 mSv/year for the effective dose is calculated for natural terrestrial gamma radiation in Costa Rica. PMID- 14750691 TI - Radon concentration in Jordanian drinking water and hot springs. AB - The radon concentration in Jordanian drinking water and hot springs has been measured using the electret-passive environmental radon monitoring method. This study maps out Jordanian water sources from the north to the south as well as the area around the capital Amman and the Jordan valley. Samples from 30 sources were collected and analysed during the period from November 2001 to June 2002. We have found that 60% of the drinking water sources have radon concentrations above the proposed US Environmental Protection Agency limit of 11 Bq l(-1); these are mostly situated in the northern region of the country. Fortunately, these sources supplied only a small number of people since they were mainly located in rural areas. Tap water in the capital Amman, where approximately 35% of the population live, has a very low radon concentration, 3.9 Bq l(-1). However, the isolated northern drinking water wells did show a radon level as high as 117 Bq l(-1). Hot springs in Jordan have a radon level ranging between 63 and 81 Bq l(-1), again mainly in the northern region. In general we conclude that Jordanian drinking water is safe as far as radon concentration is concerned with the exception of a few isolated local drinking water wells. PMID- 14750692 TI - The new ICRP memorandum and quantities in radiation protection dosimetry. PMID- 14750693 TI - Selection of healthy workers in underground metal ore miners--possible effect on leukaemia mortality. PMID- 14750694 TI - Studies of mortality of atomic bomb survivors. Report 13: Solid cancer and noncancer disease mortality: 1950-1997. PMID- 14750695 TI - COMARE statement on Green Audit occasional paper 2002/5, Cancer in Burnham on Sea North: results of the PCAH (Parents Concerned About Hinkley) Questionnaire. PMID- 14750697 TI - Actions of tumor necrosis factor-alpha on oocyte maturation and embryonic development in cattle. AB - PROBLEM: Infertility can accompany mastitis in cattle. Involvement of tumor necrosis factor-alpha (TNF-alpha) in this phenomenon is suggested by observations that circulating concentrations of TNF-alpha are elevated after intramammary infection or infusion of endotoxin. It was hypothesized that (1) TNF-alpha acts on the oocyte during maturation to decrease the percent of oocytes that cleave and develop following fertilization; (2) exposure of embryos to TNF-alpha after fertilization reduces development to the blastocyst stage; and (3) TNF-alpha increases the proportion of blastomeres that undergo apoptosis in a stage-of development dependent manner. METHOD OF STUDY: In one experiment, oocytes were matured with various concentrations of TNF-alpha and then fertilized and cultured without TNF-alpha. In another study, embryos were cultured with TNF-alpha for 8 days beginning after fertilization. Finally, embryos were collected at the two or four-cell stage (at 28-30 hr after insemination) or when > or = 9-cells (at day 4 after insemination) and cultured +/- TNF-alpha for 24 hr. The proportion of blastomeres undergoing apoptosis was then determined by the TUNEL procedure. RESULTS: Addition of TNF-alpha to maturation medium did not affect the proportion of oocytes that cleaved. However, the percent of oocytes that developed to the blastocyst stage at day 8 after insemination was reduced (P = 0.05) at all TNF alpha concentrations tested (0.1-100 ng/mL). When added during embryo culture, there was no significant effect of TNF-alpha on the proportion of oocytes that became blastocysts. In addition, TNF-alpha did not induce apoptosis in two and four-cell embryos. For embryos > or = 9-cells, however, 10 and 100 ng/mL TNF alpha increased (P < 0.05) the percent of blastomeres labeling as TUNEL-positive. CONCLUSION: TNF-alpha can have deleterious actions on oocyte maturation that compromise development of the resultant embryo. While exposure of fertilized embryos to TNF-alpha did not inhibit development to the blastocyst stage, TNF alpha increased the percentage of blastomeres undergoing apoptosis when exposure occurred for embryos > or = 9-cells. Increased blastomere apoptosis could conceivably compromise subsequent embryo survival. PMID- 14750696 TI - Influence of the murine oestrous cycle on the induction of mucosal immunity. AB - PROBLEM: To determine if the stage of oestrous cycle, at the time of immunization, affects the magnitude of mucosal and systemic immunity. METHOD OF STUDY: Female BALB/c mice were immunized with tetanus toxoid and cholera toxin by the oral, intranasal and transcutaneous routes. Groups of mice were immunized at proestrus, oestrus, postestrus and diestrus. Antibodies in serum and mucosal secretions were determined by ELISA and T cell responses by lymphocyte proliferation assay. RESULTS: Oral immunization at the oestradiol dominant stage of cycle (oestrus and proestrus) significantly enhanced TT-specific IgG and IgA levels in female reproductive tract (FRT) secretions and TT-specific IgA levels in faecal extracts. Transcutaneous immunization at diestrus enhanced TT-specific IgG in faecal extracts. TT-specific T cell proliferation is greatest following intranasal immunization at proestrus and transcutaneous immunization at diestrus, particularly in the caudal and lumbar lymph nodes draining the FRT and colon. CONCLUSIONS: Reproductive cycle-associated changes in the endogenous sex hormones oestradiol and progesterone influence the levels of vaccine-induced immunity in the FRT and distal colon following oral and transcutaneous immunization. PMID- 14750698 TI - Quantitative mRNA analysis of IL-1 gene system in human testis. AB - PROBLEM: There is a growing body of evidence that interleukins exhibit modulatory activity on development of reproductive cells. In this context, there appears to be a role for IL-1, which is also produced in human testis. We have analysed transcripts of IL-1 gene system (IL-1alpha, IL-1beta, IL-1RI, IL-1RII and IL-1RA) to evaluate the possible link between the level of gene(s) transcription and their function. METHOD OF STUDY: To determine the activity of gene transcription, a quantitative PCR with isotopic and/or nonisotopic detection was applied. RESULTS AND CONCLUSIONS: We have detected differential expression of IL-1alpha and IL-1beta genes in separate functional compartments of a male gonad. A strong expression of IL-1alpha gene in an intratubular cell fraction was shown, while the IL-1beta expression seemed to be dominant in extratubular compartment of the male gonad. Abundant amounts of IL-1RA mRNA in gametogenic cells fraction slightly higher than in interstitium have also been found. IL-1RA is the most important regulatory molecule in IL-1 system, which down-regulates activity of both interleukins. Looking more closely at gene(s) differential expression it appears that IL-1alpha can be preferentially down-regulated by IL-1RA gene in intratubular fraction while the IL-1beta, through the "false" IL-1RII receptor in the interstitium. Genes coding for both receptors (IL-1RI and IL-1RII) showed, however, relatively low levels of transcription in both studied compartments. IL 1 genes system creates a complex intragonadal environment and the function of these genes is reflected by their respective distribution in the two main functional compartments of the testis. PMID- 14750699 TI - Mutual interactions of boar seminal plasma proteins studied by immunological and chromatographic methods. AB - PROBLEM: Boar seminal plasma contains various types of proteins. Most of them belong to spermadhesins and some of them contain fibronectin type II domain. Almost all of these proteins are present under physiological conditions in aggregated forms differing in their relative molecular mass, composition, and binding properties. The study of mutual specific interactions between proteins of boar seminal plasma that could be involved in the formation of aggregated forms and most probably of sperm coating layers is the subject of our present communication. METHODS OF STUDY: Aggregated forms of boar seminal plasma proteins separated by size exclusion chromatography were analysed by Reversed-phase High performance Liquid Chromatography (RP HPLC), SDS-PAGE and immunochemical methods (ELISA and immunoblotting). Mutual interactions between proteins were investigated by size exclusion chromatography of a mixture of separated monomer proteins and affinity chromatography of boar seminal plasma on immobilized spermadhesins. RESULTS: Composition of proteins that are adsorbed from boar seminal plasma to immobilized spermadhesins corresponds to that of aggregated forms found in seminal plasma. Mutual interactions between monomer forms observed by size exclusion chromatography are in good agreement with results of affinity chromatography. An existence of interactions between spermadhesins of the AQN and AWN families and the DQH sperm surface protein, as well as between proteins involved in the formation of heterodimer porcine seminal plasma I (PSP I)/PSP II was proved. CONCLUSION: Mutual specific interactions between protein components of boar seminal plasma were shown. These interactions participate in the formation of aggregated forms of proteins in seminal plasma and probably also in the arrangement and remodelling of protein coating layers of sperm. Aggregation of seminal plasma proteins is probably an important phenomenon in the fertilization process. PMID- 14750700 TI - Characterization of spermatozoa surface antigens by antisperm antibodies and its influence on acrosomal exocytosis. AB - PROBLEM: Antisperm antibodies (ASA) are the main cause of immunological infertility, they impair sperm functions by binding to the sperm membrane. The aim of this study was to characterize highly enriched sperm membrane proteins by 2-D-electrophoresis and to identify membrane antigens binding ASA and to evaluate the influence of ASA on the acrosome reaction (AR). METHOD OF STUDY: Sperm membrane proteins were separated by 2-D-electrophoresis and antigens were identified by immunoblotting with ASA from seminal plasma samples of infertile men. The influence of ASA on the AR were observed and determined by means of flowcytometry. RESULTS: A total of 18 antigens were identified by using ASA from seminal plasma. Six of the recognized proteins were analyzed by means of mass spectrometry and peptide matching: HSP70 and HSP70-2, disulfide-isomerase-ER60, caspase-3 and two subunits of the proteasome (component-C2 and zeta-chain). ASA from seminal plasma are able to enhance the AR in donor-spermatozoa. CONCLUSION: The biochemical identification of these proteins will be helpful to understand the mechanism by which ASA impair sperm function and the fertilization process. Spermatozoa, in which the AR was prematurely induced by ASA, will not be able to fertilize anymore. PMID- 14750701 TI - Materno-fetal immunogenetic disparity: the biological basis for in vitro fertilization treatment in patients with unexplained recurrent abortion? AB - PROBLEM: The majority of women with recurrent miscarriage have no discernible cause but it has been postulated that immunologic aberrations may be the cause in most of such cases. Also, it has been stressed that deliberate modification of the maternal host defense system can improve the chances of success. We tested the hypothesis that it is possible to potentiate maternal immune functions so as to improve reproductive performance by replacing several embryos into the uterus, thus favoring the recognition of fetal antigens. METHOD OF STUDY: A total of 57 couples with three or more (mean 5.52; range 3-12) consecutive first-trimester spontaneous clinical abortions of unknown etiology were treated with in vitro fertilization (IVF) and embryo transfer for a total of 84 cycles. Patients underwent IVF after combined gonadotropin-releasing hormone agonist/gonadotropin treatment for ovarian stimulation, and up to four embryos were replaced into the uterus. RESULTS: There were 32 pregnancies (three of them after frozen-thawed embryo transfers) and 26 (81%) of them were viable gestations. Overall, patients had a previous obstetric history of 315 pregnancy losses and 15 live-born babies. Thus, the probability of having a live baby before treatment was 4.54% (95% CI, 2.78-7.36) a figure significantly lower (P < 0.0001) than that observed under IVF treatment (81%; 95% CI, 64.53-91.01). None of selected variables potentially related with the outcome of pregnancy after IVF and embryo transfer in recurrent aborters (including pre-implantation genetic diagnosis) was found to be associated with miscarriage. CONCLUSIONS: This study shows that replacement of several embryos after IVF is a useful treatment in the prevention of unexplained recurrent spontaneous abortion thus providing further evidence for immunologically modifiable pregnancy loss. PMID- 14750702 TI - Increased production of tumor necrosis factor-alpha by peritoneal fluid mononuclear cells induced by 60-kDa heat shock protein in women with minimal to mild endometriosis. AB - PROBLEM: We sought to examine the role of Hsp60, a 60-kDa stress-responsive protein (hsp60) with potent immunostimulatory properties, in the pathophysiology of endometriosis. We investigated the response of peritoneal fluid mononuclear cells (PFMCs) to hsp60 to identify differences in PFMCs between women with and without endometriosis. METHOD OF STUDY: PFMCs from women with or without endometriosis were incubated with hsp60 in vitro. The release of tumor necrosis factor (TNF)-alpha was measured by enzyme-linked immunosorbent assay (ELISA). RESULTS: The rapid secretion of TNF-alpha was observed following the exposure of PFMCs to hsp60. PFMCs from women with minimal to mild endometriosis produced greater amounts of TNF-alpha in response to hsp60 stimulation than levels seen in cells from women without endometriosis. CONCLUSION: Hsp60 may play a role in the activation of peritoneal immunity in patients with minimal to mild endometriosis. PMID- 14750703 TI - States take lead on climate change laws. PMID- 14750704 TI - Data reveal pulses of banned POPs. PMID- 14750705 TI - Examining dissolved toxic metals in U.S. estuaries. PMID- 14750706 TI - Environmental assessment of used oil management methods. AB - The 1 billion gal of used oil generated in the U.S. each year are managed in three primary ways: rerefined into base oil for reuse, distilled into marine diesel oil fuel, and marketed as untreated fuel oil. Management of used oil has local, regional and global impacts. Because of the globally distributed nature of fuel markets, used oil as fuel has localized and regional impacts in many areas. In this paper, the human health and environmental tradeoffs of the management options are quantified and characterized. The goal of this study was to assess and compare the environmental impacts and benefits of each management method in a product end-of-life scenario using a life-cycle assessment (LCA) approach. A life cycle inventory showed that 800 mg of zinc and 30 mg of lead air emissions may result from the combustion of 1 L of used oil as fuel (50-100 times that of crude derived fuel oils). As an example, up to 136 Mg of zinc and 5 Mg of lead air emissions may be generated from combustion of over 50 M gal of California generated used oil each year. While occurring elsewhere, these levels are significant (of the same magnitude as reported total stationary source emissions in California). An impact assessment showed that heavy metals-related toxicity dominates the comparison of management methods. Zinc and lead emissions were the primary contributors to the terrestrial and human toxicity impact potentials that were calculated to be 150 and 5 times higher, respectively, for used oil combusted as fuel than for rerefining or distillation. Low profits and weak markets increasingly drive the used oil management method selection toward the untreated fuel oil market. Instead, both the rerefining and distillation methods and associated product markets should be strongly supported because they are environmentally preferable to the combustion of unprocessed used oil as fuel. PMID- 14750707 TI - Will Lake Michigan lake trout meet the Great Lakes Strategy 2002 PCB reduction goal? AB - The Great Lakes Strategy 2002 establishes the long-term goal that all Great Lakes fish should be safe to eat without restriction. As an indicator of progress toward that goal, the Strategy specifies that lake trout PCBs will decline 25% from 2000 to 2007. We estimated the plausibility of achieving this near-term goal by examining a time-series of Lake Michigan lake trout PCB concentrations from 1972 to 2000. We used two different Bayesian approaches, Bayesian model averaging (BMA) and dynamic linear models (DLM), to model the trajectory of these historical data and forecast concentrations through 2007. Both approaches indicate that the probability of a 25% reduction from 2000 to 2007 is negligible. The most likely lake trout PCB declines predicted by the BMA and DLM over this time period are 6.8% and 8.9%, respectively. Our results suggest that declines in lake trout PCBs will be in the range of 5-10% assuming conditions similar to recent years. This rate of decline will be difficult to discern without adequate data collection. If sufficient data are not gathered to document further declines, then the relaxation of lake trout consumption advisories is indeed a long-term prospect. PMID- 14750708 TI - Changes in the regional emissions of greenhouse gases and ozone-depleting compounds. AB - In the wake of the Kyoto and Montreal Protocols, there is a need to verify whether policies to reduce emissions are working. We present data showing that emissions of ozone-depleting compounds, such as the chlorofluorocarbons and methyl chloroform, are decreasing from some regions of the United States but emissions of the greenhouse gases do not appear to be declining. PMID- 14750709 TI - Synthetic musk fragrances in Lake Michigan. AB - Synthetic musk fragrances are added to a wide variety of personal care and household products and are present in treated wastewater effluent. Here we report for the first time ambient air and water measurements of six polycyclic musks (AHTN, HHCB, ATII, ADBI, AHMI, and DPMI) and two nitro musks (musk xylene and musk ketone) in North America. The compounds were measured in the air and water of Lake Michigan and in the air of urban Milwaukee, WI. All of the compounds except DPMI were detected. HHCB and AHTN were found in the highest concentrations in all samples. Airborne concentrations of HHCB and AHTN average 4.6 and 2.9 ng/m3, respectively, in Milwaukee and 1.1 and 0.49 ng/m3 over the lake. The average water concentration of HHCB and AHTN in Lake Michigan was 4.7 and 1.0 ng/L, respectively. A lake-wide annual mass budget shows that wastewater treatment plant discharge is the major source (3470 kg/yr) of the synthetic musks while atmospheric deposition contributes less than 1%. Volatilization and outflow through the Straits of Mackinac are major loss mechanisms (2085 and 516 kg/yr for volatilization and outflow, respectively). Concentrations of HHCB are about one half the predicted steady-state water concentrations in Lake Michigan. PMID- 14750710 TI - Identification of long-chain perfluorinated acids in biota from the Canadian Arctic. AB - Recently it was discovered that humans and animals from various urban and remote global locations contained a novel class of persistent fluorinated contaminants, the most pervasive of which was perfluorooctane sulfonate (PFOS). Lower concentrations of perfluorooctanoate, perfluorohexane sulfonate, and heptadecafluorooctane sulfonamide have also been detected in various samples. Although longer perfluoroalkyl carboxylates (PFCAs) are used in industry and have been detected in fish following a spill of aqueous film forming foam, no studies have been conducted to examine the widespread occurrence of long-chain PFCAs (e.g., CF3(CF2)xCOO-, where x > 6). To provide a preliminary assessment of fluorinated contaminants, including PFCAs, in the Canadian Arctic, polar bears, ringed seals, arctic fox, mink, common loons, northern fulmars, black guillemots, and fish were collected at various locations in the circumpolar region. PFOS was the major contaminant detected in most samples and in polar bear liver was the most prominent organohalogen (mean PFOS = 3.1 microg/g wet weight) compared to individual polychlorinated biphenyl congeners, chlordane, or hexachlorocyclohexane-related chemicals in fat. Using two independent mass spectral techniques, it was confirmed that all samples also contained ng/g concentrations of a homologous series of PFCAs, ranging in length from 9 to 15 carbons. Sum concentrations of PFCAs (sum(PFCAs)) were lower than total PFOS equivalents (sum(PFOS)) in all samples except for mink. In mink, perfluorononanoate (PFNA) concentrations exceeded PFOS concentrations, indicating that PFNA and other PFCAs should be considered in future risk assessments. Mammals feeding at higher trophic levels had greater concentrations of PFOS and PFCAs than mammals feeding at lower trophic positions. In general, odd-length PFCAs exceeded the concentration of even-length PFCAs, and concentrations decreased with increasing chain length in mammals. PFOS and PFCA concentrations were much lower for animals living in the Canadian Arctic than for the same species living in mid-latitude regions of the United States. Future studies should continue to monitor all fluorinated contaminants and examine the absolute and relative toxicities for this novel suite of PFCAs. PMID- 14750711 TI - Characterization of airborne and bulk particulate from iron and steel manufacturing facilities. AB - Characterization of airborne and bulk particulate material from iron and steel manufacturing facilities, commonly referred to as kish, indicated graphite flakes and graphite flakes associated with spherical iron oxide particles were unique particle characteristics useful in identifying particle emissions from iron and steel manufacturing. Characterization of airborne particulate material collected in receptor areas was consistent with multiple atmospheric release events of kish particles from the local iron and steel facilities into neighboring residential areas. Kish particles deposited in nearby residential areas included an abundance of graphite flakes, tens of micrometers to millimeters in size, and spherical iron oxide particles, submicrometer to tens of micrometers in size. Bulk kish from local iron and steel facilities contained an abundance of similar particles. Approximately 60% of blast furnace kish by volume consisted of spherical iron oxide particles in the respirable size range. Basic oxygen furnace kish contained percent levels of strongly alkaline components such as calcium hydroxide. In addition, concentrations of respirable Mn in airborne particulate in residential areas and at local iron and steel facilities were approximately 1.6 and 53 times the inhalation reference concentration of 0.05 microg/m3 for chronic inhalation exposure of Mn, respectively. Thus, airborne release of kish may pose potential respirable particulate, corrosive, or toxic hazards for human health and/or a corrosive hazard for property and the environment. PMID- 14750712 TI - Occurrence of methyl triclosan, a transformation product of the bactericide triclosan, in fish from various lakes in Switzerland. AB - The bactericide triclosan and methyl triclosan, an environmental transformation product thereof, have been previously detected in lakes and a river in Switzerland. Both compounds are emitted via wastewater treatment plants (WWTPs), with methyl triclosan probably being formed by biological methylation. Passive sampling with semi-permeable membrane devices (SPMDs) showed the presence of methyl triclosan in some lakes, suggesting some potential for bioaccumulation of the compound. In this study, we report the presence of methyl triclosan in fish (white fish, Coregonus sp.; roach, Rutilus rutilus) from various lakes in Switzerland receiving inputs from WWTPs. Identification of the compound was based on mass spectral (MS) evidence including MS/MS data. The concentrations of methyl triclosan in the fish were up to 35 ng g(-1) on a wet weight basis and up to 365 ng g(-1) on a lipid basis with concentrations in a relatively narrow range for fish from the same lake (Thunersee, 4-6 ng g(-1); Zurichsee, 32-62 ng g(-1); Pfaffikersee, 43-56 ng g(-1); Greifensee, 165-365 ng g(-1), lipid basis). No methyl triclosan (<1 ng g(-1), lipid basis) was detected in fish (lake trout, Salmo trutta) from a remote lake in Sweden (Habberstjarnen) and in fish (roach) from a small lake in Switzerland with no input from WWTPs (Huttnersee, <2-<5 ng g(-1), lipid basis). The concentrations of methyl triclosan in fish correlated (r2 = 0.85) with the ratio of population in the watershed to water throughflow of the lakes (P/Q ratio), which is considered to be a measure for the domestic burden from WWTPs to a lake. Passive sampling with SPMDs confirmed the presence of methyltriclosan in lakes and a river (Zurichsee and Greifensee; Limmat) but not in a remote mountain lake (Jorisee) and in Huttnersee. The bioconcentration factor (BCF) of methyl triclosan estimated from the fish data and SPMD-derived water concentrations was in the order of 1-2.6 x 10(5) (lipid basis) and thus in the range of other persistent organic pollutants. SPMDs were found to be reliable for monitoring low concentrations of methyl triclosan in surface water. Methyl triclosan appears to be a suitable marker for WWTP-derived lipophilic contaminants in the aquatic environment and fish. PMID- 14750713 TI - Recent changes in platinum group element concentrations and osmium isotopic composition in sediments from an urban lake. AB - Automobile catalyst emissions have resulted in the occurrence of elevated Pt, Pd, and Rh concentration in the urban and roadside environment. We investigate the chronology of platinum group elements (PGE) accumulation in dated sediments from an urban lake near Boston, MA. Chronological profiles demonstrate that Pt, Pd, and Rh concentrations increased following the introduction in catalysts with accumulation rates 6-16 times larger in 1992-2002 than prior to the introduction of catalysts. Ratios of these elements closely match their ratios in catalysts, providing further evidence of an automobile source. Iridium and Ru accumulation in sediments also increased following the introduction of catalysts, and while past Os contamination is associated with leather tanning, recent changes in the isotopic composition of Os indicate another anthropogenic source for this element. The PGE have similar geochemical properties and are difficult to separate from one another, and therefore, we suggest that automobile catalyst emissions also result in increasing environmental concentrations of Ir, Ru, and Os, which occur as impurities in catalysts. An automobile catalyst source of Ir and Os is supported by elevated concentrations of these elements in a tunnel dust sample. PMID- 14750714 TI - PCDDs, PCDFs, and coplanar PCBs in albatross from the North Pacific and Southern Oceans: levels, patterns, and toxicological implications. AB - Concentrations of polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), and coplanar polychlorinated biphenyls (coplanar PCBs) were determined in five albatross species collected from the North Pacific and Southern Oceans to assess the north-south differences in residue levels, accumulation patterns, and toxic potential. Black-footed and Laysan albatrosses from the North Pacific Ocean contained higher levels of PCDD/Fs and coplanar PCBs than albatrosses from the Southern Ocean, indicating that emission sources of these contaminants were predominant in the northern hemisphere. Residue levels in albatrosses from the remote North Pacific Ocean far from the point source of pollution were comparable to or higher than those in terrestrial and coastal birds from contaminated areas in developed nations, suggesting the specific exposure and accumulation of PCDD/Fs and coplanar PCBs in albatross. The long life span and ingestion of plastic resin pellets by albatrosses could be the plausible explanations for the elevated accumulation of persistent and lipophilic contaminants including PCDD/Fs and coplanar PCBs in these birds. Relative proportions of PCDFs and coplanar PCBs in albatross were higher than those observed in birds inhabiting terrestrial and coastal areas, suggesting that these toxic chemicals may have higher transportability by air and water than PCDDs. Congener patterns of PCDD/Fs in albatross showed less variability as compared to those in terrestrial species, indicating that contamination patterns of PCDD/Fs were similar within the open ocean environment. Contributions of PCDD/Fs to total TEQs in albatrosses from the open ocean were generally lower than those in terrestrial birds, suggesting different toxic potency of PCDD/Fs and coplanar PCBs on animals inhabiting open ocean and terrestrial environment. Whereas albatrosses from southern oceans retained lower TEQ concentrations, possible adverse effects of PCDD/Fs and coplanar PCBs to black-footed and Laysan albatrosses of the North Pacific Ocean may be suspected from TEQ levels. PMID- 14750715 TI - Causes of variability in pesticide and PCB concentrations in air near the Great Lakes. AB - Data through 2001 from the Integrated Atmospheric Deposition Network (IADN) were used to investigate the causes of variability in gas-phase polychlorinated biphenyl (PCB) and pesticide concentrations measured near Lakes Michigan, Erie, and Superior. A multiple linear regression model that incorporates temperature and time was used explain the variability in the concentrations. Our approach used autocorrelation analyses of the residuals to help us determine the effectiveness of the regression. Autocorrelation plots forthe in-use pesticide lindane indicated that an agricultural application cycle was also present in the regression residual data at all sites. The addition of parameters for this effect to the regression equation accounted for, on average, 16% more of the variability in the data. Similar analyses forthe in-use pesticide endosulfan did not show an agricultural application effect. The banned compounds DDT and chlordane showed that temperature and time correctly accounted for the variability in the atmospheric concentrations of these compounds at all sites. In contrast to the other compounds, PCBs and hexachlorobenzene showed strong residual autocorrelation patterns near Lake Michigan of an unknown origin. PMID- 14750716 TI - Comparison of personal, indoor, and outdoor exposures to hazardous air pollutants in three urban communities. AB - Two-day average concentrations of 15 individual volatile organic compounds (VOCs) were measured concurrently in (a) ambient air in three urban neighborhoods, (b) air inside residences of participants, and (c) personal air near the breathing zone of 71 healthy, nonsmoking adults. The outdoor (O), indoor (I), and personal (P) samples were collected in the Minneapolis/St. Paul metropolitan area over three seasons (spring, summer, and fall) in 1999 using charcoal-based passive air samplers (3M model 3500 organic vapor monitors). A hierarchical, mixed-effects statistical model was used to estimate the mutually adjusted effects of monitor location, community, and season while accounting for within-subject and within time-index (monitoring period) correlation. Outdoor VOC concentrations were relatively low compared to many other urban areas, and only minor seasonal differences were observed. A consistent pattern of P > I > O was observed across both communities and seasons for 13 of 15 individual VOCs (exceptions were carbon tetrachloride and chloroform). Results indicate that ambient VOC measurements at central monitoring sites can seriously underestimate actual exposures for urban residents, even when the outdoor measurements are taken in their own neighborhoods. PMID- 14750717 TI - Hydrocarbons in Lake Washington sediments. A 25-year retrospective in an urban lake. AB - Aliphatic and polycyclic aromatic hydrocarbon and stable and radiocarbon isotope distributions are compared for dated cores from the 1970s and 2000 for a 25-year retrospective in Lake Washington, Seattle, WA (USA). Contamination of Lake Washington sediments by petrogenic aliphatic hydrocarbons and pyrolytic polycyclic aromatic hydrocarbons via atmospheric deposition and stormwater runoff peaked between the 1950s and 1970s and has since decreased as stormwater inputs have been reduced. Radiocarbon signatures (delta14C, per 1000) of total organic carbon decrease (increased "age") in the depth interval of highest hydrocarbon concentration. Graphitic black carbon in the year 2000 core showed a historical profile similar to that of the PAH; however high background sediments deposited before the founding of Seattle indicates a considerable nonindustrial component derived from weathering in the watershed. Unlike hydrocarbon contamination, input of terrestrial organic matter (tracked by long-chain fatty alcohols) has increased throughout the late 20th century, documenting a shift in pollutant sources away from hydrocarbons and toward anthropogenic erosion of the region's soils. PMID- 14750718 TI - Slow desorption mechanisms of volatile organic chemical mixtures in soil and sediment micropores. AB - Desorption profiles of trichloroethylene (TCE), tetrachloroethylene (PCE), and a TCE-PCE mixture were measured for three natural solids and four zeolites. Initial sorbed mass (Mi) in slow desorbing sites of natural solids and in micropores of zeolites were obtained from desorption profiles. In natural solids, Mi increases with recalcitrant organic matter content. In zeolites, Mi generally increases with decreasing micropore width and increasing micropore hydrophobicity, but the effect of hydrophobicity is stronger. In both natural solids and zeolites, competition between TCE and PCE causes Mi for each sorbate in the mixture to be less than or similar to that for each sorbate alone. Zeolite results indicate that micropore width affects this competition more than micropore hydrophobicity for the solids examined. Desorption in all solids was simulated with the radial diffusion model, either alone or coupled with the advection-dispersion equation when necessary; diffusion rate constants (D/R2) were obtained. In natural solids, mean values of D/R2 increase with decreasing recalcitrant organic matter content. In zeolites, values of D/R2 generally increase with increasing micropore width, while they are a weak function of hydrophobicity. In both natural solids and zeolites, competition between TCE and PCE causes D/R2 for each sorbate in the mixture to generally be larger than that for each sorbate alone. Zeolite results indicate that the effects of competition on D/R2 generally decrease with decreasing micropore width for the solids examined; a trend with micropore hydrophobicity is not apparent. For the three natural solids and four zeolites examined in this study, the similar effects of competition between TCE and PCE on values of Mi and D/R2 and the overlapping range of D/R2 values support the hypothesis that diffusion through hydrophobic micropores affects and may control slow mass transfer processes in the recalcitrant organic matter of natural solids. These results contribute to the fundamental understanding of slow mass transfer processes in natural solids, and they indicate that characterization of micropore width and polarity may be necessary to predict organic chemical transport and fate. PMID- 14750719 TI - Understanding the differences in Cd and Zn bioaccumulation and subcellular storage among different populations of marine clams. AB - The marine clams Mactra veneriformis were collected from three different locations in a contaminated bay in Northern China. Another species of clams Ruditapes philippinarum was collected from the same contaminated bay as well as from a relatively clean site in Hong Kong. The indices of Cd and Zn bioaccumulation (assimilation efficiency, dissolved uptake rate, and efflux rate), tissue concentration, subcellular distribution, metallothionein (MT) content, and clearance rate of the clams were subsequently quantified in these populations in the laboratory. In the two species of clams, the population with a higher Cd tissue concentration assimilated Cd and Zn more efficiently, in correlation with an increase in the Cd associated with the metallothionein-like protein (MTLP) fraction. The subcellular partitioning of Zn was similar among the different populations. The dissolved uptake rates of Cd and Zn were not influenced by the different tissue concentrations of metals in the clams. However, the clam R. philippinarum from the contaminated site reduced their Zn uptake rate constants in response to increasing Zn concentration in the water. Differences in Cd and Zn tissue concentrations had little influence on the metal efflux rate constant and the clams' clearance rate. Our results indicate that the higher Cd and Zn tissue concentrations observed in these two species may be partially caused by the high levels of metal assimilation. Populations living in contaminated environments may be able to modify their physiological and biochemical responses to metal stress, which can subsequently alter trace metal bioaccumulation to aquatic animals. The relative significance of dietary uptake and the potential trophic transfer of metals in the contaminated areas may be substantially different from those in the clean environments. PMID- 14750720 TI - Can degradation products be used as documentation for natural attenuation of phenoxy acids in groundwater? AB - In situ indicators of degradation are important tools in the demonstration of natural attenuation. A literature survey on the production history of phenoxy acids and degradation pathways has shown that metabolites of phenoxy acid herbicides also are impurities in the herbicide products, making the bare presence of these compounds useless as in situ indicators. These impurities can make up more than 30% of the herbicides. Degradation of phenoxy acids was demonstrated in microcosm experiments using groundwater and sediment contaminated with MCPP, dichlorprop, and related compounds such as other phenoxypropionic acids and chlorophenols. Field observations at two phenoxy acid-contaminated sites showed the occurrence of several impurities including metabolites in the groundwater. Neither the microcosm experiments nor the field observations verified that metabolites were actually produced or accumulated in situ. However, it was demonstrated that the impurity/parent herbicide ratios can be useful in situ indicators of degradation. PMID- 14750721 TI - In situ bioreduction of technetium and uranium in a nitrate-contaminated aquifer. AB - The potential to stimulate an indigenous microbial community to reduce a mixture of U(VI) and Tc(VII) in the presence of high (120 mM) initial NO3- co contamination was evaluated in a shallow unconfined aquifer using a series of single-well, push-pull tests. In the absence of added electron donor, NO3-, Tc(VII), and U(VI) reduction was not detectable. However, in the presence of added ethanol, glucose, or acetate to serve as electron donor, rapid NO3- utilization was observed. The accumulation of NO2-, the absence of detectable NH4+ accumulation, and the production of N2O during in situ acetylene-block experiments suggest that NO3- was being consumed via denitrification. Tc(VII) reduction occurred concurrently with NO3- reduction, but U(VI) reduction was not observed until two or more donor additions resulted in iron-reducing conditions, as detected by the production of Fe(II). Reoxidation/remobilization of U(IV) was also observed in tests conducted with high (approximately 120 mM) but not low (approximately 1 mM) initial NO3- concentrations and not during acetylene-block experiments conducted with high initial NO3-. These results suggest that NO3(-) dependent microbial U(IV) oxidation may inhibit or reverse U(VI) reduction and decrease the stability of U(IV) in this environment. Changes in viable biomass, community composition, metabolic status, and respiratory state of organisms harvested from down-well microbial samplers deployed during these tests were consistent with the conclusions that electron donor additions resulted in microbial growth, the creation of anaerobic conditions, and an increase in activity of metal-reducing organisms (e.g., Geobacter). The results demonstrate that it is possible to stimulate the simultaneous bioreduction of U(VI) and Tc(VII) mixtures commonly found with NO3- co-contamination at radioactive waste sites. PMID- 14750722 TI - Modeling tetracycline antibiotic sorption to clays. AB - Sorption interactions of three high-use tetracycline antibiotics (oxytetracycline, chlortetracycline, tetracycline) with montmorillonite and kaolinite clays were investigated undervaried pH and ionic strength conditions. Sorption edges were best described with a model that included cation exchange plus surface complexation of zwitterion forms of these compounds. Zwitterion sorption was accompanied by proton uptake, was more favorable on acidic clay, and was relatively insensitive to ionic strength effects. Calcium salts promoted oxytetracycline sorption at alkaline pHs likely by a surface-bridging mechanism. Substituent effects among the compounds in the tetracycline class had only minor effects on sorption edges and isotherms under the same solution pH and ionic strength conditions. At low ionic strength, greater sorption to montmorillonite than kaolinite was observed at all pHs tested, even after normalizing for cation exchange capacity. These results indicate that soil and sediment sorption models for tetracyclines, and other pharmaceuticals with similar chemistry, must account for solution speciation and the presence of other competitor ions in soil or sediment pore waters. PMID- 14750723 TI - Factors affecting the distribution of the rate of carbon uptake by forests in South Korea. AB - The biomass of forests in South Korea has significantly increased during the last 30 years because of a national reforestation project and forest management. Despite the high potential of this biomass for sequestering atmospheric carbon dioxide, little effort has been devoted to identifying the factors affecting the rate of carbon uptake by forests in Korea. Recently, we reported that Korean forests have a higher carbon uptake rate (1.5 Mg C ha(-1) year(-1)) than those of North America, Europe, and China. In this study, as a follow-up to that work, we examine the distributions of total forest carbon, carbon density, and carbon uptake rate at the province and subprovince levels in Korea and elucidate the relationships between forest and climatic variables within these distributions. The provinces can be classified into three distinct groups according to their carbon uptake rate and forest age class: group A (Gyeonggi, Chungbuk, Chungnam), group B (Gyeongbuk, Gyeongnam, Jeonbuk, Jeonnam), and group C (Gangwon, Jeju). When all forest and climatic variables are considered, the provinces Gangwon and Jeju in group C are found to belong to distinct groups. The rate of carbon uptake in each province is not significantly correlated to most forest and climatic variables but is highly correlated to forest age class. A multivariate statistical analysis also supports our conclusion that forest age class is the major factor affecting the current distribution of the rate of carbon uptake in Korea. We conclude that for several decades Korean forests will have a high capacity for sequestering carbon dioxide. PMID- 14750724 TI - Characterization of sorption mechanisms of VOCs with organobentonites using a LSER approach. AB - To fully utilize the sorption traits of organobentonites to control volatile organic compounds (VOCs) pollution, the sorption mechanisms of VOCs with organobentonites need to be understood adequately. The sorption of VOCs as vapors to a typical organobentonite, modified with cetyltrimethylammonium bromide (CTMAB bentonite), was characterized using a linear solvation energy relationship (LSER) of the type log Kc = c + rR2 + s pi2H + a sigma(alpha2)H + b sigma(beta2)H + l log L16. The fitted LSER equation, log Kc = 0.434 + 0.968R2 - 0.0886pi2H + 2.170sigma(alpha2)H + 1.611sigma(beta2)H + 0.417 log L16, was obtained by a multiple regression of the partition coefficients of 22 probe solutes against the solvation parameters of the solutes. The coefficients of the LSER equation show that CTMAB-bentonite is a sorbent with nonsignificant dipolarity/polarizability, interacts with solutes partly through pi-/n-electron pairs, behaves both as hydrogen-bond donor and hydrogen-bond acceptor, and can interact with solutes by cavity/dispersion interactions. The related terms in LSER suggest that the potential factors governing the sorption of VOCs onto CTMAB-bentonite are dispersion interactions, hydrogen-bond acidity interactions, hydrogen-bond basicity interactions, and pi-/n-electron interactions. The dispersion interaction is recognized to be the predominant parameter for most solutes, whereas the contributions of the other parameters depend on specific solutes. The derived LSER equation successfully predicted the VOC partition coefficients and the selectivity of CTMAB-bentonite for the VOCs. The relationship between LSER and adsorption/partition model was compared. The classification of sorption mechanisms by LSER goes on the molecular interaction types between sorbate and sorbent, and classification by adsorption/partition model goes on the property difference among various components of sorbent. The LSER approach coupled with inverse gas chromatography (IGC) is a comparatively simple and reliable tool to rapidly characterize the sorption mechanism of VOCs with solid sorbents such as CTMAB-bentonite, and may potentially be applied to the design of an organoclay sorbent for control of VOCs. PMID- 14750725 TI - Metal ion sorption and desorption on zeolitized tuffs from the Nevada Test Site. AB - Because of the hundreds of nuclear weapon tests conducted on the Nevada Test Site (NTS) during the Cold War, the migration of radionuclides and contaminants is a potential concern. The mobility of these compounds and our ability to remediate contaminated sites are controlled by sorption and desorption processes, which depend frequently on the nature of the contaminant, the mineralogy of the site, and the geochemical conditions. The sorption and desorption behavior of strontium (Sr) and lead (Pb), two metal cations with different chemistries, commonly found on nuclear test sites were studied. Strontium showed pH-independent and ionic strength-dependent sorption, consistent with ion exchange processes at permanent charge sorption sites. The sorption uptake of Sr increased with decreasing ionic strength of background solution. Strontium desorption from the adsorbents was enhanced by increased background electrolyte concentration and was a function of background electrolyte composition. The fractional uptake of Pb was higher, compared to that of Sr, and was only pH dependent at the highest ionic strength used (1.0 M). This pH-dependent sorption behavior, consistent with formation of surface complexes at amphoteric surface hydroxyl sites or formation of surface precipitates, could explain the decreased Pb desorption, compared to that of Sr, especially at increased background electrolyte concentrations. Under conditions typical for the groundwater at the NTS (I = 0.003 M, pH = 8.0), both Pb and Sr are expected to bind strongly on tuffs with composition similar to the zeolitized tuffs used in this study. Any increase in the dissolved ion concentration of the groundwater, however, may result in, at least partial, release of Sr and enhanced Sr mobility. PMID- 14750726 TI - Structure effect on the interaction of phenylurea herbicides with model biomembrane as an environmental mobility parameter. AB - During recent years, intensive use of herbicides has raised increasing concern mainly due to their massive pollution of the environment. As these herbicides are directly or indirectly toxic to a wide range of organisms, their potential for contaminating soil, surface water, and groundwater makes these xenobiotics of special interest from a health and environmental point of view. Knowledge of the mechanisms by which they exert their toxic effects is becoming a need. Because of the herbicides' lipophilicity, a possible site of interaction in the cell is represented by biomembranes. The interaction of four herbicides, difenoxuron, diuron, linuron, and metoxuron, with model membranes constituted of dimyristoylphosphatidylcholine multilamellar vesicles was investigated by the differential scanning calorimetry technique. The aim was to study the effects exerted by an increasing amount of the examined compounds on thermotropic behavior of the model phospholipid membranes and to correlate the obtained results with structural features of the herbicides due to their environmental mobility. Among the herbicides studied, linuron is the most effective in perturbing the ordinate structure of vesicles forming phospholipids, whereas metoxuron is the least effective and the others exert an intermediate effect. Linuron exerts its effect both on the transition temperature of the gel to the liquid crystalline phase and on the enthalpy change. Difenoxuron, diuron, and metoxuron cause a change in the transition temperature but have an insignificant effect on the enthalpy change. The calorimetric results, correlated with the structural features of the herbicides, are consistent with their partition coefficient, log K(ow), suggesting that the more hydrophobic compound character causes a greater liposolubility and consequential cellular absorption with more effectiveness on the membrane order. PMID- 14750727 TI - Photoinduced oxidation of Hg0(aq) in the waters from the St. Lawrence estuary. AB - The oxidation of volatile aqueous Hg0 in aquatic systems may be important in decreasing the fluxes of Hg out of the water column. Using incubations of natural samples from the St. Lawrence River, we examined some of the parameters that control this oxidation. Hg0 was found to be chiefly mediated by UV radiation since (i) "dark" oxidation was not found to be statistically significant; (ii) visible light induced a significant but slow photooxidation (k = 0.09 h(-1)); and (iii) visible + UV radiation led to a faster photooxidation (k = 0.6-0.7 h(-1)), mainly because of UV-A induced reactions. Doubling UV irradiation did not increase the reaction rate of Hg0 photooxidation in natural water samples, indicating that some factor other than photon flux was rate limiting and suggesting that the reaction involves intermediate photoproduced oxidant(s). The addition of methanol, a *OH scavenger, decreased Hg photooxidation rates by 25% in brackish waters and by 19% in artificial saline water containing semiquinones, indicating that *OH may be partly responsible for Hg0 oxidation. Photooxidation rates were not affected by oxygen concentrations and did not decrease when samples were heat-sterilized, treated with chloroform, or filtered prior to exposure to light. PMID- 14750728 TI - Emission, fate and effects of soluble silicates (waterglass) in the aquatic environment. AB - Soluble silicates, commercially known as waterglass, are among the largest volume synthetic chemicals in the world. Silicon from waterglass is rapidly transformed to the biologically active orthosilicic acid (referred to as dissolved silicate). This paper aims to assess the impact of waterglass on the aquatic environment in Western Europe. The emission to surface waters from the four most relevant application areas, household detergents, pulp and paper production, water and wastewater treatment, and soil stabilization, is estimated to be ca. 88-121 kton of SiO2 per year. This is a small fraction (<2%) of the estimated total amount of dissolved silicate transported by rivers to the oceans. Locally, increases in dissolved silicate concentration will decrease the ratios of N:Si and P:Si, which could influence phytoplankton species composition and favor the growth of diatoms over other groups of algae. Significant adverse effects in aquatic ecosystems are not expected. PMID- 14750729 TI - Phenylpyrazole insecticide fipronil induces male infertility in the estuarine meiobenthic crustacean Amphiascus tenuiremis. AB - Copepods are the most abundant arthropods on earth and are often the most important secondary producers in estuarine/marine food webs. The new GABA (gamma aminobutyric acid)-disrupting insecticide fipronil (FP) induces unique sex specific reproductive dysfunction in male meiobenthic copepods, leading to trans generational population depression at environmentally realistic concentrations (0.63 microg/L). Using a newly developed 96-well microplate lifecycle bioassay, more than 700 individual Stage-I juveniles were reared to adulthood in as short as 12 days in only 200 microL of control (CTL) or 0.63 microg-FP/L seawater solution. Individual virgin male: female pairs were then cross-mated for all possible combinations within and across rearing treatments and allowed to mate for an additional 12 days in CTL or 0.63 microg-FP/L solution. FP at 0.63 microg/L caused no significant lethality to any mating combinations but evoked 73% or 89% inhibition of reproduction when FP-reared males were mated with either a control- or FP-reared female in FP solution, respectively. In contrast, when CTL-reared males were mated with FP-reared females in FP solution, there was no difference in reproductive success compared to FP-free controls. When FP-reared males were mated with either female group in FP-free solution, these mating pairs displayed a 3-day delay in time to brood sac extrusion but ultimately did reproduce. As fipronil (1) has a high K(ow), (2) is persistent in sediments where meiobenthic copepods live, and (3) has been detected in estuarine waters >0.7 microg/L, it may pose high risk to copepod production in estuarine systems. PMID- 14750730 TI - Correlation equation for predicting single-collector efficiency in physicochemical filtration in saturated porous media. AB - A new equation for predicting the single-collector contact efficiency (eta0) in physicochemical particle filtration in saturated porous media is presented. The correlation equation is developed assuming that the overall single-collector efficiency can be calculated as the sum of the contributions of the individual transport mechanisms--Brownian diffusion, interception, and gravitational sedimentation. To obtain the correlation equation, the dimensionless parameters governing particle deposition are regressed against the theoretical value of the single-collector efficiency over a broad range of parameter values. Rigorous numerical solution of the convective-diffusion equation with hydrodynamic interactions and universal van der Waals attractive forces fully incorporated provided the theoretical single-collector efficiencies. The resulting equation overcomes the limitations of current approaches and shows remarkable agreement with exact theoretical predictions of the single-collector efficiency over a wide range of conditions commonly encountered in natural and engineered aquatic systems. Furthermore, experimental data are in much closer agreement with predictions based on the new correlation equation compared to other available expressions. PMID- 14750731 TI - Accumulation and toxicity of cadmium in the aquatic oligochaete Tubifex tubifex: a kinetic modeling approach. AB - Heavy metal pollution is a serious threat to ecosystem functioning. Different approaches have been developed to relate the exposure of heavy metals to their accumulation and toxicity. One approach is to relate metal toxicity to the concentrations of the metals in the whole body or a specific target tissue instead of the external exposure concentrations. To test the usefulness of this approach, the relationship between cadmium exposure, accumulation, and toxicity was investigated using an oligochaete worm and kinetic modeling. The uptake and elimination of cadmium by the aquatic oligochaete Tubifex tubifex from the aqueous phase was studied as function of time at different exposure concentrations using both radioactive and non-radioactive cadmium. A two compartmental pharmacokinetic model was constructed and parametrized by fitting the model to the measured cadmium body concentrations during exposure to different cadmium concentrations. The uptake rate constants were dependent on the cadmium exposure concentration, and this relation could be well-described by incorporation of Michaelis-Menten type uptake kinetics. The toxicity of cadmium was analyzed by determining the lethal exposure concentration associated with a mortality of 50% (LC50) at different time points. LC50 values decreased with increasing exposure time reaching the incipient lethal level after 15 d. Critical body concentrations (CBC) associated with 50% mortality were calculated by combining the model-predicted pharmacokinetic parameters and the measured LC50 values. The predicted mean CBC (0.32 micromol/g wet weight +/- 0.02) was in good agreement with the experimentally obtained CBC for cadmium found in T. tubifex (0.37 micromol/g wet weight +/- 0.07) and appeared to be independent of exposure time and exposure concentration. Our results show that a pharmacokinetic modeling approach provides a tool to link metal exposure to availability, accumulation, and toxicity under variable exposure scenarios taking into account the kinetics of the processes. PMID- 14750732 TI - Multilinear model for spatial pattern analysis of the Measurement of Haze and Visual Effects project. AB - A multilinear model was developed for the analysis of the spatial patterns and possible sources affecting haze and its visual effects in the southwestern United States. The data from the project Measurement of Haze and Visual Effects (MOHAVE) collected during the late winter and mid-summer of 1992 at the monitoring sites in four states (i.e., California, Arizona, Nevada and Utah) were used in the study. The three-way data array was analyzed by a four-product-term model. This study makes a direct effort to include wind patterns as a component in the model in order to obtain the information of the spatial patterns of source contributions. The solution is computed using the conjugate gradient algorithm with applied non-negativity constraints. For the winter data set, reasonable solutions contained six sources and six wind patterns. The analysis of summer data required seven sources and seven wind patterns. The ME results are compared to the prior single-species empirical orthogonal function analysis results--and prior work describing the transport pathways. PMID- 14750733 TI - Global source attribution for mercury deposition in the United States. AB - A multiscale modeling system that consists of a global chemical transport model (CTM) and a nested continental CTM was used to simulate the global atmospheric fate and transport of mercury and its deposition over the contiguous United States. The performance of the CTMs was evaluated against available data. The coefficient of determination (r2) for observed versus simulated annual mercury wet deposition fluxes over North America was 0.50 with average normalized error and bias of 25% and 11%, respectively. The CTMs were used to conduct a global source attribution for selected receptor areas. Three global emission scenarios were used that differed in their distribution of background emissions among direct natural emissions and re-emissions of natural and anthropogenic mercury. North American anthropogenic sources were calculated to contribute only from 25 to 32% to the total mercury deposition over the continental United States. At selected receptors, the contribution of North American anthropogenic emissions ranges from 9 to 81%; Asian anthropogenic emissions were calculated to contribute from 5 to 36%; natural emissions were calculated to contribute from 6 to 59%. PMID- 14750734 TI - Geographic sensitivity of fine particle mass to emissions of SO2 and NOx. AB - An air quality model, URM-1ATM, was used to investigate tendencies in fine particle (PM2.5) species in response to changes in SO2 and NOx emissions in the eastern United States. The model employed the decoupled direct method (DDM) to estimate sensitivities without the need for multiple model runs for different emissions species and geographic regions. The baseline for sensitivities was emissions projected to 2010. Principal geographic regions investigated were east of the Mississippi River, although the contribution of a region to the immediate west of the river was also included in the study. Sensitivities to emissions changes from point sources (SO2 and NOx) and low-level sources (NOx) were computed. PM2.5 species examined were sulfate, organic carbon, and nitrate as well as total fine mass. Results for the midwest, mid-Atlantic, and southeast regions indicated that those regions affect their own aerosol levels the most. Aerosols in the northeast were most strongly linked to emissions from the midwest and mid-Atlantic regions. In general, midwest emissions had the most influence of any region on other regions. In addition, the southeast was relatively isolated, having the least influence outside itself and being least affected by neighboring regions. Sulfate was the species most sensitive to emission changes. Finally, the largest potential relative sensitivities of sulfate and organic aerosols, along with PM2.5 mass, to emissions changes were usually modeled to occur outside those areas computed to experience the highest aerosol levels. PMID- 14750735 TI - Validated measurements of the uranium isotopic signature in human urine samples using magnetic sector-field inductively coupled plasma mass spectrometry. AB - Increased interest in measuring uranium isotope ratios in environmental samples (biological materials, soils, dust particles, water) has come from the necessity to assess the health impact of the use of depleted uranium (DU) based ammunitions during recent military conflicts (e.g., Gulf war, Kosovo) and from the need to identify nondeclared nuclear activities (nuclear safeguards). In this context, very important decisions can arise which have to be based on measurement data of nondisputable uncertainty. The present study describes the certification to 2.5% (k = 2) relative combined uncertainty of n(235U)/n(238U) at ultralow uranium levels (approximately 5-20 pg g(-1)) in human urine samples. After sample decomposition and matrix separation, the isotope ratios were measured by means of a single-detector magnetic sector-field inductively coupled plasma mass spectrometry instrument fitted with an ultrasonic nebulizer. Correction for mass discrimination effects was obtained by means of the certified isotopic reference material IRMM-184. The analytical procedure developed was validated in three complementary ways. First, all major sources of uncertainty were identified and propagated together following the ISO/GUM guidelines. Second, this quality was controlled with a matrix matching NUSIMEP-3 sample (approximately 0.06-0.7% difference from certified). Third, the instrumental part of the procedure was proven to be reproducible from the confirmation of the results obtained for three samples remeasured 7 months later (approximately 1.5% difference). The results obtained for 33 individuals indicated that none seemed to have been exposed to contamination by DU. PMID- 14750736 TI - In-situ measurements of Cu in an estuarine environment using a portable spectrophotometric analysis system. AB - Application of a portable in-situ spectrophotometric analysis system for the measurement of Cu in estuarine environments is described in this work. Our spectrophotometric elemental analysis system (SEAS) used for in-situ observations of Cu concentrations is capable of fully autonomous or user-controlled operations. The optical cells used in SEAS systems are flexible liquid core waveguides (LCWs) with optical path lengths as long as 5 m. The 1-m waveguide used in the present study provided a 3.0 nM detection limit and a 5.0% relative standard deviation for a 25 nM copper sample. Analysis times range between 1 and 5 min, allowing for acquisition of data on scales appropriate to the highly dynamic biogeochemical nature of copper in the coastal environment. Field deployments of SEAS-Cu in Tampa Bay, FL, showed low Cu concentrations near the mouth of the estuary (3-4 nM), with elevated concentrations (approximately 25 nM) in anthropogenically impacted regions of the bay (e.g., marinas and areas adjacent wastewater treatment plants). Transect data between Tampa Bay and a deep water harborage exhibited copper concentrations ranging between 5 and 50 nM. PMID- 14750737 TI - Investigation of the wetting behavior of coal tar in three phase systems and its modification by poloxamine block copolymeric surfactants. AB - The removal of dense nonaqueous phase liquid mixtures (DNAPLs) from rocks and subsurface soils is an ongoing remedial challenge. Very often the wetting preferences of the system are not altered by exposure to the DNAPL. However, there are systems where the wetting properties of the solid phase have been altered from strongly water wetting by exposure to the DNAPL. In these cases some technique is necessary for reducing the work of adhesion between the DNAPL and the mineral surface. The focus of this report is the problems posed by coal tar in unconsolidated sands. It is shown that coal tar can alter the wetting properties of quartz, the principal component of sands, and is thus capable of adhering to the surface. In this investigation the ability of several members of the poloxamine family of polymeric surfactants to aid in the removal of coal tar from sand was evaluated. The poloxamines are tetrafunctional block copolymeric surfactants, which contain four poly(ethylene oxide)-block-poly(propylene oxide) chains joined to a central ethylenediamine moiety via the nitrogen atoms. Contact angle measurements of coal tar on a quartz surface immersed in aqueous surfactant solution and the interfacial tension between coal tar and aqueous surfactant solution have been measured. Coal tar/water interfacial tensions are reduced to values in the region of 2 mN m(-1) at surfactant concentrations of approximately 0.1 w/v %. Poloxamine surfactant impact on the static contact angle is more complex. In some cases the polymeric surfactants alter the wetting behavior from strongly water wetting to weakly water wetting. However, other poloxamines appear to have little if any impact on the contact angle, which remains strongly water wetting. The foregoing measurements have then been used to calculate the work of adhesion of the coal tar to quartz and the results qualitatively compared with the concentration of surfactant solution required to visually demonstrate the complete de-adhesion of coal tar to the quartz. It is shown that at surfactant concentrations below the critical micelle concentration (cmc) of the surfactant, the work of adhesion can be reduced sufficiently to ensure complete removal of coal tar from both quartz and sand. PMID- 14750738 TI - Identification of dynamic leaching kinetics of stabilized, water-soluble wastes. AB - A one-dimensional diffusion model based on Fick's second law with a non-zero surface concentration at the solid-solution interface was developed to calculate effective calcium and sulfate diffusion coefficients of composites placed in saltwater. A regression method was used to identify the leaching kinetics. The regression method decomposes the stabilized PG leaching processes into diffusion, surface wash-off, and immediate and long-term precipitation. The immediate surface precipitation of both calcium and sulfate ions occurred only in three of the PG composite combinations. The effective diffusion coefficients of calcium (2.58-4.68 x 10(-13) m2 s(-1)) and sulfate (2.77-5.02 x 10(-13) m2 s(-1)) obtained from the regression method are similar to those obtained from methods of cumulative flux and daily flux associated with the simple diffusion model, provided that the leaching processes do not deviate significantly from that of the diffusion. The ratio (1.13) of effective sulfate to calcium diffusion coefficients obtained using the regression analysis is statistically consistent with the theoretical value (1.31), which further justifies the regression method. The research also implies that the leaching processes of calcium and sulfate ions stop after a certain period of time (300-900 d for calcium and 80-170 d for sulfate) and that the precipitations of calcium and sulfate affect the leaching processes. The regression method can be used to identify the leaching mechanisms and to predict the long-term stability of the stabilized wastes. PMID- 14750739 TI - Hydrogen and carbon isotope fractionation during anaerobic biodegradation of aromatic hydrocarbons--a field study. AB - The aquifer of a former manufactured gas plant site, highly contaminated by dissolved monocyclic, heterocyclic, and polycyclic aromatic hydrocarbons, was studied to evaluate the applicability of carbon and hydrogen isotope fractionation to prove ongoing biodegradation of these compounds even in complex aquifer settings. The loss of toluene, o-xylene, p,m-xylene, and 2 methylnaphthalene was accompanied by a considerable carbon isotope fractionation. Additionally, a strong 2H enrichment in residual o-xylene was detected. All isotope fractionations observed could be related to established biochemical degradation mechanisms, each involving a C-H bond cleavage in the rate determining step. In contrast, other compounds such as 1-methylnaphthalene, methylbenzofuran, and acenaphthene exhibited a uniform stable carbon isotope composition. However, a decrease in concentration for these compounds was observed in the flowpath of the aquifer. High threshold concentrations of acenaphthene downgradient indicate that this contaminant is, if at all, only marginally biodegraded. Detailed analyses of xylenes provided support that compound specific isotope analyses and subsequent application of the Rayleigh model may provide a valuable basis to distinguish between different biodegradation mechanisms as well as dissolution processes in heterogeneous aquifers. PMID- 14750740 TI - Combined application of stable carbon isotope analysis and specific metabolites determination for assessing in situ degradation of aromatic hydrocarbons in a tar oil-contaminated aquifer. AB - To evaluate the intrinsic bioremediation potential in an anoxic tar oil contaminated aquifer at a former gasworks site, groundwater samples were qualitatively and quantitatively analyzed by compound-specific isotope analysis (CSIA) and signature metabolites analysis (SMA). 13C/12C fractionation data revealed conclusive evidence for in situ biodegradation of benzene, toluene, o xylene, m/p-xylene, naphthalene, and 1-methylnaphthalene. In laboratory growth studies, 13C/12C isotope enrichment factors for anaerobic degradation of naphthalene (epsilon = -1.1 +/- 0.4) and 2-methylnaphthalene (epsilon = -0.9 +/- 0.1) were determined with the sulfate-reducing enrichment culture N47, which was isolated from the investigated test site. On the basis of these and other laboratory-derived enrichment factors from the literature, in situ biodegradation could be quantified for toluene, o-xylene, m/p-xylene, and naphthalene. Stable carbon isotope fractionation in the field was also observed for ethylbenzene, 2 methylnaphthalene, and benzothiophene but without providing conclusive results. Further evidence for the in situ turnover of individual BTEX compounds was provided by the presence of acetophenone, o-toluic acid, and p-toluic acid, three intermediates in the anaerobic degradation of ethylbenzene, o-xylene, and p xylene, respectively. A number of groundwater samples also contained naphthyl-2 methylsuccinic acid, a metabolite that is highly specific for the anaerobic degradation of 2-methylnaphthalene. Additional metabolites that provided evidence on the anaerobic in situ degradation of naphthalenes were 1-naphthoic acid, 2 naphthoic acid, 1,2,3,4-tetrahydronaphthoic acid, and 5,6,7,8-tetrahydronaphthoic acid. 2-Carboxybenzothiophene, 5-carboxybenzothiophene, a putative further carboxybenzothiophene isomer, and the reduced derivative dihydrocarboxybenzothiophene indicated the anaerobic conversion of the heterocyclic aromatic hydrocarbon benzothiophene. The combined application of CSIA and SMA, as two reliable and independent tools to collect direct evidence on intrinsic bioremediation, leads to a substantially improved evaluation of natural attenuation in situ. PMID- 14750741 TI - Combined bioaugmentation and biostimulation to cleanup soil contaminated with high concentrations of atrazine. AB - We developed a joint bioaugmentation and biostimulation approach for the clean up of soil contaminated with high (168.7 and 337.4 microg g(-1)) concentrations of the herbicide atrazine (2-chloro-4-(ethylamino)-6-isopropylamino-s-triazine). Pseudomonas sp. strain ADP (P. ADP) was used for bioaugmentation (approximately 10(7) cells g(-1) soil), and citrate (concentration range 5.8-40 mg g(-1) soil) and succinate (6.2-30.8 mg g(-1)) were used for biostimulation. The study soil had indigenous potential for atrazine mineralization (54.4 +/- 2% of 168.7 microg g(-1) mineralized after 67 day), but rapid mineralization only took place after a prolonged acclimation phase (approximately 28 days). Inoculation with P. ADP alone resulted in a limited improvement in mineralization (e.g., 30.6 +/- 1% mineralization of 168.7 microg g(-1) of atrazine in inoculated soil cf. < 0.5% in noninoculated in 7 days). Quantification of surviving numbers of P. ADP revealed a 10-fold decline from initial levels. However, bioaugmentation together with citrate or succinate biostimulation markedly increased P. ADP cell survival and atrazine mineralization (e.g., addition of 11.6 mg g(-1) of citrate increased mineralization of 337.4 microg g(-1) of atrazine from < 2 to 79.9 +/- 1% in 13 days). A critical parameter in determining the extent of atrazine mineralization by P. ADP was C(s):N(atz) (soluble carbon to atrazine nitrogen ratio): C(s):N(atz) > 40 was required for maximal atrazine mineralization. We suggest our observations may be used as a framework for rational bioremediation of field soils contaminated with atrazine. PMID- 14750742 TI - Electrochemical hydrodehalogenation of 2,4-dibromophenolin paraffin oil using a solid polymer electrolyte reactor. AB - A new technology for remediation of halogenated organics-oil systems, which can cause serious environmental problems, has been demonstrated using the electrochemical hydrodehalogenation of 2,4-dibromophenol (DBP) in paraffin oil in a solid polymer electrolyte reactor. The reactor has been evaluated in terms of cathode materials and structure and the ratio of the cathode surface area to the solution volume. A cathode of titanium minimesh with a palladium electrocatalyst produced by electrodeposition was particularly effective. Current efficiencies of up to 85% and percentage of DBP removal of up to 62%, space-time yields of up to 7.6 kg DBP m(-3) h(-1), and energy consumption as low as 1.6 kW h (kg of DBP)(-1) were achieved. The reactor showed stable operation for periods of up to 170 h. The results demonstrated that electroreduction could be an alternative technology to electrooxidation forthe treatment of wastes and toxic halogenated compounds, making the process simpler in comparison to electrooxidation. PMID- 14750743 TI - Mobilizing particles in a saturated zone during air sparging. AB - The mobilization of soil particles changes the porosity of saturated zone during air sparging. Soil porosity is shown to be correlated with soil electrical resistivity. This study performs porosity-resistivity tests to establish the relationship between porosity and resistivity of quartz sand. Experiments, involving a large sandbox to simulate the saturated zone, are then performed to compare the resistivity of compacted sand before air injection with that after air injection. The relevant data enable the mobilization of quartz sand particles to be quantified. Results of the experiments indicate the mobilization of sand particles and an increase in porosity directly proportional to the rate at which air is injected. Besides, a layer of fine-grained particles covered the compacted sand at the upper boundary of sandbox after each air injection experiment. This is direct evidence that finer particles were transported upward during air sparging. Two methods were applied to verify the results of this study. The first verification method indicated that changes in porosity increased directly proportional to the air injection rate, which is consistent with shear theory. The other validation method indicated that the mass of sand in the tank did not change after air sparging, which indicates that the resistivity-porosity method is unbiased. PMID- 14750744 TI - Methylmercury formation in a wetland mesocosm amended with sulfate. AB - This study used an experimental model to evaluate methylmercury accumulation when the soil of a constructed wetland is amended with sulfate. The model was planted with Schoenoplectus californicus and designed to reduce wastestream metals and metal-related toxicity. The soil was varied during construction to provide a control and two sulfate treatments which were equally efficient at overall mercury and copper removal. After an initial stabilization period, methylmercury concentrations in porewater were up to three times higher in the sulfate-treated porewater (0.5-1.6 ng/L) than in the control (<0.02-0.5 ng/L). Mean percent methylmercury was 9.0% in the control with 18.5 and 16.6% in the low- and high sulfate treatments, respectively. Methylmercury concentrations measured in mesocosm surface water did not reflect the differences between the control and the sulfate treatments that were noted in porewater. The mean bulk sediment methylmercury concentration in the top 6 cm of the low-sulfate treatment (2.33 ng/g) was significantly higher than other treatment means which ranged from 0.96 to 1.57 ng/g. Total mercury in sediment ranged from 20.8 to 33.4 ng/g, with no differences between treatments. Results suggest that the non-sulfate-amended control was equally effective in removing metals while keeping mercury methylation low. PMID- 14750746 TI - ECG of the month. Oxymoron. Accelerated idioventricular rhythm. PMID- 14750745 TI - Shall the last be first? Only if we have the will to improve. PMID- 14750747 TI - Radiology case of the month. Radiologic evaluation of a bone tumor in a child. Benign osteochondroma with surrounding bursal sac. PMID- 14750748 TI - Pathology case of the month. Tumor of the cauda equina. Paraganglioma of the cauda equina. PMID- 14750749 TI - Lymphatic malformations. AB - Lymphatic malformations are developmental anomalies of the lymphatic system that can range in size and symptomatology and are usually diagnosed in early childhood. To follow is a discussion of the proposed etiology, diagnostic methods, and categorization, as well as debated methods of treatment of these malformations. PMID- 14750750 TI - Essential thrombocytosis: diagnostic and treatment dilemmas. AB - Essential thrombocytosis is one of the chronic myeloproliferative disorders that includes polycythemia vera, chronic myelogenous leukemia, and agnogenic myeloid metaplasia. Despite established diagnostic criteria and greater than a quarter century of study, there are still several diagnostic and management dilemmas that plague researchers and clinicians alike. We present a case of essential thrombocytosis in a patient with multiple sequelae. PMID- 14750751 TI - Cardiovascular disease in patients infected with the human immunodeficiency virus. AB - Worldwide, infection with the human immunodeficiency virus (HIV) is increasing. At the same time, new treatments allow patients to live longer. Consequently, cardiovascular manifestations, most of which occur relatively late in the course of the infection, are becoming more frequent. Pericardial effusion, the most common cardiovascular manifestation of HIV infection, usually is small and causes no hemodynamic compromise or symptoms. It does, however, augur a grim prognosis, as do other cardiovascular conditions associated with the infection: myocarditis, dilated cardiomyopathy, pulmonary arterial hypertension, cardiac lymphoma, and Kaposi's sarcoma of the heart. Highly active antiretroviral therapy (HAART), especially when incorporating protease inhibitors, greatly improves overall outlook in these patients, but appears not only to cause a lipodystrophic syndrome, but to accelerate atherosclerotic cardiovascular disease by inducing glucose intolerance, frank diabetes mellitus, hypertriglyceridemia, hypercholesterolemia, increased lipoprotein (a), and decreased HDL cholesterol. Recent ongoing prospective trials also are showing an association of HAART with increased coronary artery disease and myocardial infarction. PMID- 14750752 TI - Education does pay off: pneumococcal vaccine screening and administration in hospitalized adult patients with pneumonia. AB - Streptococcus pneumoniae-associated infections are an important cause of hospitalization and mortality in high-risk and elderly patients. Even in the setting of appropriate therapy, the case fatality rate of invasive pneumococcal disease in the elderly may approach 40%. Since approximately 40,000 people die annually from pneumococcal-associated disease, it represents a substantial target for vaccine-preventable, bacterial fatalities. The 23-valent pneumococcal polysaccharide vaccine has proven consistently effective in preventing invasive pneumococcal disease. Despite its endorsement by numerous specialty societies, the pneumococcal vaccine is underutilized in the inpatient setting. In a recent report of quality indicators for Medicare beneficiaries, the percentage of Medicare beneficiaries in Louisiana admitted with pneumonia who were screened or received the pneumococcal vaccination prior to discharge was only 4%, the lowest percentage in the United States. The Louisiana State University-New Orleans Internal Medicine Department and its house staff embarked upon a retrospective study to determine its baseline pneumococcal vaccination or screening rates for all patients with pneumonia on its inpatient services at the The Medical Center of Louisiana in New Orleans from July 2000 through June 2001. From July 2001 through June 2002 an intensive educational intervention concentrating on the indications and benefits of pneumococcal vaccination was directed toward the Louisiana State University Internal Medicine house staff assigned to the inpatient service. Retrospective analysis for pneumococcal vaccine screening and administration of charts of all patients with pneumonia on the LSU Medicine service from July 2001 through June 2002 was performed in order to determine the effects of the intervention. Data from the pre-educational intervention period revealed a baseline pneumococcal vaccine screening or administration rate of 11% for all patients with pneumonia on the LSU Internal Medicine inpatient service. During the one-year intervention period, the pneumococcal vaccine screening or administration rate increased to 71%, a clinically and statistically significant increase (p-value < 0.0001). Data targeting patients 65 years of age and older revealed a baseline pneumococcal vaccine screening or administration rate of 10% for patients with pneumonia on the LSU Internal Medicine inpatient service which increased to 82% during the one year educational intervention (p-value < 0.0001). House officer scores (possible range 0-100) on a questionnaire assessing their understanding of the indications and benefits of pneumococcal vaccination were significantly higher after the educational intervention compared to before the intervention (means +/- standard deviations, 68 +/- 9 vs. 59 +/- 10, p < 0.0001). The findings from this study highlight the importance of education in increasing compliance with widely-accepted practice guidelines such as pneumococcal vaccine screening or administration in patients hospitalized with pneumonia. PMID- 14750753 TI - Louisiana State University School of Public Health. AB - On July 1, 2003, the Louisiana State University School of Public Health became the 6th School at LSU Health Sciences Center in New Orleans joining the Schools of Medicine, Nursing, Dentistry, Allied Health Professions, and Graduate Studies. Its primary mission will be to improve the health of the people of Louisiana and their families and communities through education, research, and community service, with special emphasis on the health and healthcare of underserved people and on the industries of importance to the state. Its programs and faculty will be developed primarily in collaboration with other components of the LSU system. PMID- 14750754 TI - Risk stratification in mild head injury patients: the head injury predictive index. AB - BACKGROUND: Mild Head Injury has classically been defined as patients with a Glasgow Coma Scale (GCS) of 13 to 15. While most of these patients do well, some suffer serious outcomes. The objective of this study was to identify the group at high risk for poor outcome. METHOD: Analysis of 255 consecutive patients who presented to the Medical Center of Louisiana at New Orleans with GCS from 13 to 15 was performed. A new Head Injury Predictive Index (HIPI) based on Focal Neurological Signs, the verbal response and eye-opening components of the GCS, and CT findings was used to stratify patients into "high" and "low" risk groups. RESULTS: There were 10 in-hospital decompensations and 7 poor outcomes at discharge. The study shows that the HIPI was predictive of both poor outcomes and in-patient decompensations. CONCLUSION: When compared to GCS, the inclusion of computed tomographic and focal neurological data gives the HIPI more power in predicting poor outcome and in-hospital decompensations in the mild head injury group. PMID- 14750755 TI - The historical odyssey of cardiac transplantation. AB - Often taken for granted, the journey leading to the therapeutic reality of cardiac transplantation was paved by many obstacles and a myriad of successes rooted primarily in the basic principle of perseverance. One cannot pretend to understand the sensational evolution of cardiac transplantation without a look back at the historical accomplishments concurrently achieved across a variety of disparate disciplines in medicine, surgery, and immunology that heralded contemporary outcomes. This historical review will delineate landmark events in the field of cardiac transplantation, a procedure once considered a "fantastic speculation for the future." PMID- 14750756 TI - Nonpenetrating deep sclerectomy with the use of adjunctive mitomycin C. AB - BACKGROUND AND OBJECTIVE: To determine whether adjunctive use of mitomycin C (MMC) would increase the success rate of primary nonpenetrating deep sclerectomy with collagen implant. PATIENTS AND METHODS: Twenty-six patients (26 eyes) with primary open-angle glaucoma uncontrolled with maximally tolerated medical therapy were randomly assigned to undergo nonpenetrating deep sclerectomy and collagen implant, either with (13 eyes) or without (13 eyes) 0.3 mg/mL of adjunctive MMC for 3 minutes. Intraocular pressure (IOP), number of glaucoma medications, and visual acuity were assessed before and 12 and 24 months after surgery. RESULTS: The mean age of the study patients was similar in both groups (MMC = 68.1 +/- 8 years, control = 65.8 +/- 6.8 years). At the 12- and 24-month follow-up visits, the mean IOP with or without medications was lower in the MMC group than in the control group (15.6 +/- 3.5 vs 17.2 +/- 3.9 mm Hg at 12 months and 15.8 +/- 5.6 vs 17.8 +/- 2.8 mm Hg at 24 months, respectively). The IOP with or without medications significantly decreased after surgery in both groups (P < .05). Twelve months after surgery, the IOP decreased by 48% in the MMC group and by 35% in the control group. At the end of follow-up (24 months), the IOP decreased by 48% in the MMC group and by 32% in the control group (P = .01). The mean number of glaucoma medications decreased after surgery, there was no significant difference in complications (hyphema and suprachoroidal hemorrhage), and visual acuity was unchanged throughout the study in both groups. CONCLUSION: The use of MMC in nonpenetrating deep sclerectomy with collagen implant is safe and improves surgical results. PMID- 14750757 TI - Combined phacoemulsification, intraocular lens implantation, and scleral buckling surgery for cataract and retinal detachment. AB - BACKGROUND AND OBJECTIVE: To evaluate the effects of simultaneous surgery for cataract and retinal detachment. PATIENTS AND METHODS: Six patients with a diagnosis of retinal detachment accompanied by dense cataracts precluding fundus visualization were enrolled in this study. A triple procedure involving phacoemulsification, posterior chamber intraocular lens insertion, and scleral buckling was performed in all six cases. RESULTS: A clear intraoperative view of the fundus was obtained in all cases and retinal breaks were identified in five cases. The retina was successfully reattached in all six cases, five of which showed improvement in visual acuity. CONCLUSION: Combined cataract and scleral buckling surgery can be performed safely. This procedure spares the patient repeat surgery and may improve the visual outcome by clearly identifying the breaks and avoiding delays in detachment repair. PMID- 14750758 TI - Interferon alpha-2a for proliferative diabetic retinopathy after complete laser panretinal photocoagulation treatment. AB - BACKGROUND AND OBJECTIVE: The aim of this study was to investigate the effect of interferon alpha-2a, an angiogenesis inhibitor, on eyes with active neovascularization after complete laser panretinal photocoagulation treatment. PATIENTS AND METHODS: Eight patients with active neovascularization persisting for 6 months or more after completion of full panretinal photocoagulation were included in the study. All patients were treated with subcutaneous injections of 6 million international units of interferon alpha-2a, 3 times a week, for an average period of 10 months. Visual acuity, contrast sensitivity, blood tests, fundus photographs, fluorescein angiography, and physical examination were performed periodically. The main outcome measures were visual acuity and extent of neovascularization as assessed by fundus photography and fluorescein angiography. RESULTS: The 5 men and 3 women (mean age, 60 years) had a mean duration of diabetes of 19 years. The average study follow-up was 42.2 +/- 8.7 weeks. Visual acuity and extent of neovascularization improved or remained stable in 7 patients. In none of the patients was there progression of neovascularization, but in 1 patient it could not be assessed due to vitreous hemorrhage. Most patients had malaise during the first weeks of treatment, but none of the patients suffered from nonreversible side effects associated with interferon alpha-2a. CONCLUSION: This pilot study provides evidence that interferon alpha-2a might have a role in the regression of proliferative diabetic retinopathy and that further investigation is warranted. PMID- 14750759 TI - Indocyanine green staining for the triple corneal procedure. AB - BACKGROUND AND OBJECTIVE: In the triple corneal procedure, successful completion of capsulorhexis is of the utmost importance. Another use of indocyanine green dye for better visualization of the anterior lens capsule of mature cataract during the triple corneal procedure is described. PATIENTS AND METHODS: Four consecutive patients (mean age, 69.5 years) with both mature cataracts and corneal opacity underwent the triple corneal procedure. After trephination of the recipient cornea, the anterior capsule of the lens was stained with indocyanine green. A continuous curvilinear capsulorhexis (CCC) was performed, after which conventional triple corneal procedures were followed. RESULTS: In all four cases, this technique markedly improved visualization of the lens capsule and resulted in successful and easy manipulation of the CCC and subsequent removal of residual lens cortex. CONCLUSIONS: Staining of the anterior capsule of mature cataract in the triple corneal procedure clearly defines the border of the capsule, thus allowing easy and complete execution of CCC. PMID- 14750760 TI - Large spot endolaser for retinal photocoagulation and transvitreal thermotherapy. AB - BACKGROUND AND OBJECTIVE: Diode laser transpupillary thermotherapy can be used for the treatment and choroidal tumors. A large spot endolaser probe was developed for endophotocoagulation and transvitreal thermotherapy. The aim of this study was to assess the probe's clinical performance for photocoagulation and transvitreal thermotherapy combined with macular elevation. MATERIALS AND METHODS: Endolaser with a divergent beam giving a 3 mm spot size on the retina was developed. Vitrectomy was performed on ten eyes of five Dutch Belted rabbits. Threshold for photocoagulation was determined for the diode (810 nm) laser in four eyes. In another six eyes, the retina was detached with balanced salt solution prior to laser irradiation. The tissue effects were monitored clinically and with photography and microscopy. RESULTS: Threshold photocoagulation was induced with 150 mW power, for 1 minute, in attached retina. Following retinal detachment, no coagulation occurred at 200 mW (suprathreshold) power for 1 minute. CONCLUSIONS: A large spot endolaser probe may be useful for endophotocoagulation. It generates more gradual photocoagulation than standard endolaser probes, giving a more controllable reaction over a larger area. This may be useful for treatment of large areas of the retina when treating retinal detachments or ischemic retinopathies. When detached retina was irradiated, no retinal damage occurred at suprathreshold power. This development may allow for intraoperative transvitreal thermotherapy of choroidal neovascularization while protecting the retina with intentional intraoperative detachment of the retina. PMID- 14750761 TI - Effect of disinsertion of rectus eye muscles on aqueous humor composition in rabbits. AB - BACKGROUND AND OBJECTIVES: To evaluate aqueous humor composition and biochemical analysis in a rabbit model for anterior segment ischemia. MATERIALS AND METHODS: Six rabbits underwent bilateral disinsertion of the four rectus eye muscles. One day later, aqueous humor (0.1 to 0.2 mL) was withdrawn from the anterior chamber and the concentrations of electrolytes, glucose, and lactate were determined. Similar assessments were performed in 8 eyes of 4 control rabbits that were not operated on. RESULTS: Sodium and calcium concentrations (mean +/- standard deviation) were significantly higher in the study group than in the control group (142 +/- 2.9 vs 138 +/- 2 mmol/L, P = .002, and 1.4 +/- 0.1 vs 1.3 +/- 0 mmol/L, P = .005, respectively). Potassium concentrations were significantly lower in the study group than in the control group (4.1 +/- 0.5 vs 4.5 +/- 0.1 mmol/L; P = .02). The mean glucose level in the study group was 108.4 mg/dL, which is comparable to the published normal value of 108 mg/dL, and the mean lactate level was 124 mg/dL, which is significantly higher than the published normal value of 78 mg/dL (P = .001). CONCLUSIONS: Bilateral disinsertion of the four rectus muscles in rabbits resulted in a breakdown of the blood-aqueous barrier, as reflected and measured by an electrolyte imbalance. These experimental results may be useful in the future in searching for methods of manipulating the ischemic response. There is currently no indication to perform anterior chamber taps in humans for that purpose. PMID- 14750763 TI - Determination of excimer laser ablation rates of corneal tissue using wax impressions of ablation craters and white-light interferometry. AB - BACKGROUND AND OBJECTIVE: Efforts to better understand the processes of corneal ablation are aided by a means to precisely measure corneal tissue ablation rates. MATERIALS AND METHODS: A white-light interferometer was used to measure the depth of argon fluoride laser ablation craters and wax impressions of the ablation craters. The technique was validated using polymethylmethacrylate (PMMA), and was then used to measure the ablation rate of bovine corneal tissue. RESULTS: The ablation rate was measured as 0.47 microm/pulse from direct measurement of the PMMA ablation craters and as 0.48 microm/pulse from measurement of the corresponding wax impressions. The ablation rate of bovine corneal tissue was found to be 0.90 microm/pulse. CONCLUSIONS: Wax impressions provide a valid representation of ablation craters. The ablation depth of corneal tissue and PMMA was found to increase linearly with increasing number of laser pulses, and the slope of the regression lines provided a measure of the laser ablation rate. PMID- 14750762 TI - Quantitative lacrimal scintigraphy after dacryocystorhinostomy. AB - BACKGROUND AND OBJECTIVE: To evaluate lacrimal drainage in patients who have undergone successful dacryocystorhinostomy using quantitative dacryoscintigraphy. PATIENTS AND METHODS: This study included 72 eyes of 72 patients. There were 26 males and 46 females, with a mean age of 39.6 years (range, 8 to 67 years). The patients were subdivided into three groups. Group 1 consisted of 35 eyes with external dacryocystorhinostomy, group 2 consisted of 15 eyes with endoscopic dacryocystorhinostomy, and group 3 consisted of 22 eyes with conjunctival dacryocystorhinostomy with a Jones tube. There was no epiphora in patients and the lacrimal drainage systems were patent by irrigation. The other normal eyes of the patients were evaluated as controls. RESULTS: In groups 1 and 2, the mean T(1/2) values for the palpebral aperture were lower than those for normal eyes, and this difference was statistically significant (P < .01). In group 3, no significant difference was detected in the mean T(1/2) values for the palpebral aperture. CONCLUSIONS: Tear flow was slower in patients who had had external and endoscopic dacryocystorhinostomy, but conjunctival dacryocystorhinostomy did not affect T(1/2) values for the palpebral aperture. PMID- 14750764 TI - Optical coherence tomography findings in solar retinopathy. AB - Solar retinopathy is characterized by involvement of the outer retinal layers. Optical coherence tomography may help delineate the retinal injury site in vivo. Optical coherence tomography was performed in three eyes of two patients with solar retinopathy. All three eyes demonstrated a characteristic excavation in the outer hyperreflective layer corresponding to the retinal complex. PMID- 14750765 TI - Enophthalmos as initial manifestation of occult, mammogram-negative carcinoma of the breast. AB - A patient with enophthalmos was found to have occult metastatic breast carcinoma to the orbit despite normal findings on breast examination and mammogram. Enophthalmos can be a sign of serious systemic disease and should be appropriately investigated. Despite radiologically interpreted normal orbital and breast imaging, the clinician should rule out underlying subclinical malignancy and consider orbital biopsy. PMID- 14750766 TI - Photodynamic therapy with verteporfin for subfoveal choroidal neovascular membranes in highly myopic eyes after laser in situ keratomileusis. AB - The authors describe the management of subfoveal choroidal neovascular membranes in highly myopic eyes after laser in situ keratomileusis with photodynamic therapy. Five cases of choroidal neovascular membrane after laser in situ keratomileusis for the correction of myopia (mean, 13.3 diopters; range, -8.00 to -16.25 diopters) treated with single or multiple sessions of photodynamic therapy with verteporfin are presented. Two cases had improved visual acuity (2 to 5 lines) after photodynamic therapy, two cases remained the same, and one case lost 4 lines of visual acuity. Photodynamic therapy with verteporfin seems to increase the chance of stabilizing or improving vision in patients with subfoveal choroidal neovascular membrane after laser in situ keratomileusis in highly myopic eyes. PMID- 14750767 TI - Choroidal neovascularization following laser in situ keratomileusis. AB - Two cases of choroidal neovascularization occurring after laser in situ keratomileusis (LASIK) are presented from two separate centers. In the first case, an extrafoveal choroidal neovascular membrane, apparently associated with age-related macular degeneration, occurred 3 months after a LASIK procedure was performed on a 64-year-old man with hyperopia. Argon laser treatment and subsequent photodynamic therapy were performed and resulted in stabilization of vision. The second case involved neovascularization 3 weeks following a LASIK procedure for myopia, apparently associated with the myopia. No treatment was recommended and the vision stabilized at 20/50. Although the cause is not clear, careful preoperative macular evaluation and attention to symptoms that might herald the presence of choroidal neovascularization are recommended for patients undergoing LASIK. PMID- 14750768 TI - Dislocation of a scleral-fixated posterior chamber intraocular lens into the anterior chamber associated with pseudophakic bullous keratopathy. AB - A 48-year-old patient with a scleral-fixated posterior chamber intraocular lens had dislocation of the intraocular lens into the anterior chamber and associated pseudophakic bullous keratopathy. The patient underwent intraocular lens extraction and partial penetrating keratoplasty. Because an anterior chamber intraocular lens is easier to implant and has fewer complications, surgeons may want to consider this as the first choice for treatment. If the scleral-fixated posterior chamber intraocular lens is used, it must be implanted properly and meticulously to avoid complications. PMID- 14750769 TI - Pars plana vitrectomy foreign body extraction assisted with a 24-gauge needle tunnel. AB - A 54-year-old man presented with corneal laceration, traumatized iris, small vitreous hemorrhage, lens opacity, and a 13-mm intraocular foreign body embedded in the retina. Pars plana vitrectomy and argon laser photocoagulation were performed. Using intraocular forceps, the object was forced into a 24-gauge needle that was inserted through the sclerotomy. The foreign body and the needle were removed from the eye. Phacoemulsification with foldable intraocular lens implantation in the ciliary sulcus was performed. Seven days after the surgery, the patient's visual acuity was 20/20 with spectacle correction of -3.5 diopters. The use of a needle tunnel during pars plana vitrectomy should be considered in similar cases of large, irregular intraocular foreign bodies to avoid severe and irreversible damage to the adjoining tissues. PMID- 14750770 TI - Free iris cyst in the anterior chamber. AB - A 50-year-old man had a free iris cyst in the anterior chamber that was treated by puncturing using an Nd:YAG laser. Following laser treatment, the cyst wall settled in the anterior chamber angle and no complications were encountered during 4 months of follow-up. PMID- 14750771 TI - Self-illuminated contact lens for peripheral vitreous surgery. AB - A modified self-retaining plano-concave contact lens for visualization of the ora serrata and anterior vitreous during vitreous surgery is described. A groove to hold a light pipe is made in the standard self-retaining, plano-concave, one piece acrylic lens with footplates. A drop of viscoelastic material between the lens and the cornea holds the lens in place by suction. The groove (1 mm in depth and diameter) supports the tip of the endoillumination probe held by an assistant. This illuminates the area while the surgeon indents the sclera with the free hand, facilitating quick and well-controlled vitreous base dissection. This system offers a simple, effective, and quick method for illumination of the ora serrata and anterior vitreous in aphakic and pseudophakic eyes during vitrectomy. PMID- 14750772 TI - Photodynamic therapy with verteporfin for choroidal neovascularization complicating angioid streaks. PMID- 14750773 TI - Work and housework conditions and depressive symptoms among married women: the importance of occupational status. AB - Using the American Changing Lives Survey, a nationally representative sample of adults residing in the United States, this research examines housewives' subjective evaluations of their housework and the subjective evaluations of paid employment among three groups of married women--professionals, sales-clerical, and service-blue collar wives. A major goal was to assess the usefulness of disaggregating employed women by occupational status. Depressive symptoms were regressed on five work conditions--autonomy, physical and time demands, boredom, and feeling appreciated--along with sociodemographic characteristics. The results indicate professional wives report fewer symptoms than homemakers, sales clerical, and service-blue collar wives. Differences between professionals and homemakers are largely accounted for by professional women's more advantaged economic position. Nonprofessional employed women are more depressed than professionals even when their disadvantaged working conditions are controlled. We discuss the findings in view of research on the stress of combining full-time employment with homemaking and argue that balancing these two roles may be more difficult for some employed women than for others. PMID- 14750774 TI - The relationships of hassles and uplifts to experience of health in working women. AB - Swedish women are more ill than men are, often explained by women's heavier total workload. A balanced pattern of daily occupations is believed to promote health. The aim of this study was to investigate the relationship of aspects of the pattern of daily occupations and lifestyle factors to working mothers' health and well-being, and whether control influenced any relationships. One hundred working, cohabiting mothers took part in semi-structured interviews targeting health and well-being, control, lifestyle variables, and hassles and uplifts in the pattern of daily occupations. All variables were dichotomised according to a median cut and subjected to logistic regression analyses. Working more and having a university diploma were found to be risk factors for experiencing more hassles. Risk factors for fewer uplifts were having more than two children and fewer leisure occupations. Experiencing less control constituted a risk of low self rated health and with an additional high level of hassles impacted on the experience of well-being. PMID- 14750775 TI - Social change, migration and sexual health: Chilean women in Chile and Australia. AB - Cultural beliefs, norms and values regarding sexuality and gender roles forge people's sexual behaviour and understanding of sexual health risk. Acknowledging a person's cultural background is a key challenge for the promotion of sexual health programs and strategies for the prevention of sexually transmitted diseases (STDs) and HIV/AIDS. This challenge acquires larger dimensions when health promotion programs are directed towards migrant communities. This article examines narratives about past and present life experiences of Chilean women living in Australia and Chilean women in Chile. We inquire about social changes and exposure to education women experienced in their own country and in Australia and the ways in which migrant women define and articulate their experiences in relation to sexual health prevention. In comparing these experiences, we raise a number of questions about sexual health promotion and programs, including the prevention of STDs and HIV/AIDS targeted to specific migrant communities in Australia. Very few sexual health policies and strategies in Australia take into account the impact that the social and cultural background of migrants, social changes and the 'settlement process' has on the cultural construction of gender identity of migrants in the new country. We propose that these cultural constructs are key in the formulation of migrants' beliefs and attitudes towards sexuality and sexual health. We suggest that there is a need to build effective and culturally appropriate sexual health promotion and prevention strategies that build upon the social and cultural background and the present and past life experiences of migrant women and men. PMID- 14750776 TI - Stigma, disclosure, and psychological functioning among HIV-infected and non infected African-American women. AB - HIV is on the rise among African-American women. AIDS-related stigma plays an important role in the lives of HIV-infected and non-infected African-American women. Among HIV-infected women, the decision to disclose HIV seropositive status is likely affected by perceived stigma. The first purpose of the study is to examine perceived AIDS-related stigma over a six year period and across two groups of African-American women: HIV-infected and non-infected. The second purpose of the study examines whether disclosure of HIV seropositive status moderates the relationship between stigma and psychological functioning. Participants were 98 HIV-infected and 146 non-infected African-American women, between the ages of 18 and 50. Data were collected at four points across six years. Results indicated that HIV-infected women perceived a significantly higher level of AIDS-related stigma than non-infected women at all four assessments. Perceptions of stigma did not significantly change over time for the entire sample or within either HIV group. Among HIV-infected women, as the level of perceived stigma increased, the level of disclosure and psychological functioning decreased. Regarding the hypothesized moderating relationship, at high, but not low, levels of disclosure, the relationship between stigma and distress was significant. Implications for health professionals' work with HIV-infected African-American women around the issue of disclosure and stigma are discussed. PMID- 14750777 TI - Feminism and women's health professions in Ontario. AB - Historically, prevailing gender ideologies were an important element in both the exclusionary strategies employed by male occupational groups and the countervailing responses by female groups. The way in which evolving gender ideologies, and feminism in particular, influence the continuing struggle for greater status and recognition by female professions, however, remains to be fully explored. In this paper, we examine the impact and the role of feminism and feminist ideologies within three female professional projects: nursing, dental hygiene and midwifery in Ontario. We argue that feminism provides an ideology of opposition that enables leaders in these professions to battle against professional inequalities by laying bare the gender inequalities that underlie them. Framing their struggles in feminist terms, female professions also seek recognition for the uniquely female contribution they make to the health care division of labour. At the same time, there exists a tension between ideals of feminism and ideals of professionalism, that has the potential to undermine female professional projects. PMID- 14750778 TI - Physicians as violence prevention activists--a qualitative study. AB - Few studies have examined the challenges facing physician activists: health care providers who engage in unpaid, non-clinical work to effect change in social issues pertaining to public health. We conducted focus groups with 19 health care providers active in violence prevention; data were analyzed using qualitative methods. Five themes emerged: (1) personal experience had generated participants' activism; (2) physicians believed they were uniquely qualified as violence prevention activists; (3) violence prevention inside the health care setting often overshadowed outside activism; (4) they feared being overwhelmed by demands of activism; and (5) they felt isolated and valued networking, especially locally, to relieve isolation. Findings illustrate the complex demands of violence prevention work on today's busy physicians. PMID- 14750779 TI - Developing capacity for integrated trauma-related behavioral health services for women: start-up costs from five community sites. AB - Violence against women is a growing public health concern. Many community agencies are interested in expanding their service delivery capacity to better meet these needs, but little information is currently available about the costs of implementing comprehensive services for women who have experienced various types of trauma. This article responds to this need by presenting findings from a study of start-up costs incurred by five regionally-diverse sites in the Women, Co-Occurring Disorders, and Violence Study (Women's Study) during the initial two years of a 5-year federal demonstration grant. Using a cost-finding approach that included project-related expenditures without regard to the source of funding, information was obtained with a structured data collection instrument that was completed during in-person interviews with local project staff. Phase 1 start-up project costs ranged from 0.6 million dollars to almost 1.2 million dollars per site. Of the five sites, start-up costs were lowest in the two mixed urban/suburban settings, and highest in the two rural settings, where few of the project-related services had been in place prior to grant funding. Implications for public health interventions are highlighted. PMID- 14750780 TI - Active site mapping of 2-deoxy-scyllo-inosose synthase, the key starter enzyme for the biosynthesis of 2-deoxystreptamine. Mechanism-based inhibition and identification of lysine-141 as the entrapped nucleophile. AB - A key enzyme in the biosynthesis of clinically important aminoglycoside antibiotics including neomycin, kanamycin, gentamicin, etc. is 2-deoxy-scyllo inosose synthase (DOIS), which catalyzes the carbocycle formation from d-glucose 6-phosphate to 2-deoxy-scyllo-inosose (DOI). To clarify its precise reaction mechanism and crucial amino acid residues in the active site, we took advantage of a mechanism-based inhibitor carbaglucose-6-phosphate (pseudo-dl-glucose, C-6 P) with anticipation of its conversion to a reactive alpha,beta-unsaturated carbonyl intermediate. It turned out that C-6-P clearly showed time- and concentration-dependent inhibition against DOIS, and the molecular mass of the resulting modified-DOIS with C-6-P was 160 mass units larger than that of native DOIS. Thus, the expected alpha,beta-unsaturated intermediate appeared to trap a specific nucleophilic group in the active site through the Michael-type 1,4 addition. The covalently modified amino acid residue was determined to be Lys-141 by means of enzymatic digestion and subsequent LC/MS and LC/MS/MS of the digest. Also discussed are the role of Lys-141 in the substrate recognition and the reaction pathway and comparison with evolutionary related dehydroquinate synthase. PMID- 14750781 TI - Design, synthesis, and evaluation of 9-D-ribitylamino-1,3,7,9-tetrahydro-2,6,8 purinetriones bearing alkyl phosphate and alpha,alpha-difluorophosphonate substituents as inhibitors of tiboflavin synthase and lumazine synthase. AB - Lumazine synthase and riboflavin synthase catalyze the last two steps in the biosynthesis of riboflavin, an essential metabolite that is involved in electron transport processes. To obtain structural probes of these two enzymes, as well as inhibitors of potential value as antibiotics, a series of ribitylpurinetriones bearing alkyl phosphate and alpha,alpha-difluorophosphonate substituents were synthesized. Since the purinetrione ring system and the ribityl hydroxyl groups can be alkylated, the synthesis required the generation of these two moieties in protected form before the desired alkylation reaction could be carried out. These substances were designed as intermediate analogue inhibitors of lumazine synthase that would bind to its phosphate-binding site. All of the compounds were found to be effective inhibitors of both Bacillus subtilis lumazine synthase as well as Escherichia coli riboflavin synthase. Molecular modeling of the binding of 3 (1,3,7,9-tetrahydro-9-D-ribityl-2,6,8-trioxopurin-7-yl)propane 1-phosphate provided a structural explanation for how these compounds are able to effectively inhibit both enzymes. Interestingly, the enzyme kinetics of these new compounds in comparison with the parent purinetrione demonstrated unexpectedly that the phosphate and phosphonate substituents contributed negatively to the binding. A possible explanation for these effects on lumazine synthase would be that the inorganic phosphate in the assay buffer competes with the substituted purinetriones for binding to the enzyme. This would be consistent with the observed increase in K(m) of the 3,4-dihydroxybutanone-4-phosphate substrate from 5.2 microM in Tris buffer or from 6.7 microM in MOPS buffer to 50 microM in phosphate buffer when tested on Bacillus subtilis lumazine synthase. However, when tested in Tris buffer vs Mycobacterium tuberculosis lumazine synthase, three of the phosphate inhibitors displayed inhibition constants in the 4-5 nM range, indicating that they are much more potent than the parent purinetrione. Under these conditions, the phosphate moieties of the inhibitors do contribute positively to their binding. The alpha,alpha-difluorophosphonate analogue, which is expected to have enhanced metabolic stability relative to the phosphates, was also found to be an inhibitor of Mycobacterium tuberculosis lumazine synthase with a K(i) of 60 nM. PMID- 14750782 TI - The first direct evaluation of the two-active site mechanism for chitin synthase. AB - Chitin synthase polymerizes UDP-GlcNAc to form chitin (poly-beta(1,4)-GlcNAc) and is essential for fungal cell wall biosynthesis. The alternating orientation of the GlcNAc residues within the chitin chain has led to the proposal that chitin synthase possesses two active sites. We report the results of the first direct test of this possibility. Two simple uridine-derived dimeric inhibitors are shown to exhibit 10-fold greater inhibition than a monomeric control, consistent with the presence of two active sites. This observation has important implications for the development of antifungal agents, as well as the understanding of polymerizing glycosyltransferases. PMID- 14750783 TI - Synthesis and biological activity of enantiomers of antitumor irofulven. AB - Stereoselective synthesis of (-)-irofulven has been achieved by cycloaddition of (R)-5-chloro-5-methyl-2-cyclopentenone to the 1,3-dipolar intermediate from 1 acetyl-1-(diazoacetyl)cyclopropane. The enantiomer, (+)-irofulven, was prepared in a similar way starting with (S)-5-chloro-5-methyl-2-cyclopentenone. (+) Irofulven was 5 to 6 times less toxic than (-)-irofulven to adenocarcinoma (MV 522) cells. PMID- 14750784 TI - The first broad application of alkynyl sulfides as dienophiles in cobalt(I) catalyzed Diels-Alder reactions. AB - The cobalt(I)-catalyzed Diels-Alder reaction of nonactivated aryl alkynyl sulfides with acyclic 1,3-dienes generates dihydroaromatic vinyl sulfides under very mild reaction conditions, and these products can be oxidized with mild oxidants to the corresponding diaryl sulfides in good overall yields. The steric and electronic effects of substituents on the aryl, as well as on the alkynyl, moieties of the aryl alkynyl sulfide are discussed. While the cobalt catalyst system is quite efficient in converting alkynyl sulfides to the Diels-Alder adducts, the transformation of the corresponding aryl alkynyl sulfoxides and sulfones under similar mild reaction conditions gave only moderate yields of the desired adducts. PMID- 14750785 TI - A high-spin and durable polyradical: poly(4-diphenylaminium-1,2 phenylenevinylene). AB - A purely organic, high-spin, and durable polyradical molecule was synthesized: It is based on the non-Kekule- and non-disjoint design of a pi-conjugated poly(1,2 phenylenevinylene) backbone pendantly 4-substituted with multiple robust arylaminium radicals. 4-N,N-Bis(4-methoxy- and -tert-butylphenyl)amino-2 bromostyrene 5 were synthesized and polymerized with a palladium-phosphine catalyst to afford the head-to-tail-linked polyradical precursors (1). Oxidation of 1 with the nitrosonium ion solubilized with a crown ether gave the aminium polyradicals (1(+)()) which were durable (half-life > 1 month) at room temperature in air. A high-spin ground state with an average S = (4.5)/2 for 1a(+) was proved even at room temperature by magnetic susceptibility, magnetization, ESR, and NMR measurements. PMID- 14750786 TI - Multigram synthesis of well-defined extended bifunctional polyethylene glycol (PEG) chains. AB - A series of novel, well-defined, unsymmetrical poly(ethylene glycol) chains of the type X(OCH(2)CH(2))(n)()Y (where X = protecting group; Y = nucleofuge or a different protecting group; n = 3, 6, 9, 12, 15, 18, and 24) were prepared in high yields by applying orthogonal protecting groups. The purity of the compounds was fully verified by elemental and high-resolution mass spectrometry analyses. PMID- 14750787 TI - Resonance energies of the allyl cation and allyl anion: contribution by resonance and inductive effects toward the acidity and hydride abstraction enthalpy of propene. AB - Density functional theory was employed to calculate the acidities and hydride abstraction enthalpies of propene (3) and propane (4), along with their vinylogues (5 and 6, respectively). The same reaction enthalpies were calculated for the propene vinylogues in which the terminal vinyl group was rotated perpendicular to the rest of the conjugated system (7). The contribution by resonance and inductive effects toward the acidity and hydride abstraction enthalpy of each vinylogue of 5 (n = 1-3) was computed and extrapolated to n = 0 (the parent propene system). The resonance energies of the allyl cation and anion were determined to be about 20-22 and 17-18 kcal/mol, respectively. Comparisons are made to resonance energies calculated using other methodologies. PMID- 14750788 TI - Highly stereoselective synthesis of aristeromycin through dihydroxylation of 4 aryl-1-azido-2-cyclopentenes. AB - Dihydroxylation of 4-aryl-1-azido-2-cyclopentenes 6, in which an aryl group is used as a synthetic equivalent of CH(2)OH, was studied to improve the low to moderate stereoselectivity previously reported for cyclopentenes 3 possessing CH(2)X and nitrogen atom-containing groups. 2-Furyl, Ph, and p-MeOC(6)H(4) groups were chosen as the aryl groups. Compounds 6a-c possessing such aryl groups were prepared by CuCN-catalyzed reaction between 2-cyclopentene-1,4-diol monoacetate 9 and the corresponding Grignard reagents followed by substitution of the hydroxyl group with (PhO)(2)P(=O)N(3). The desired diols 7a-c were obtained with higher selectivities of >7:1 when dihydroxylation of 6a-c was carried out at 0 degrees C with OsO(4) (catalyst) and NMO in a mixed solvent of MeCN, THF, t-BuOH, and H(2)O. Among them, the furyl compound recorded the highest selectivity of 14:1. The furyl and azido groups on diol 7a were converted into hydroymethyl and adeninyl groups, respectively, to produce acetonide 2, which upon hydrolysis affords aristeromycin 1. PMID- 14750789 TI - Experimental studies of the 13C NMR of iodoalkynes in Lewis-basic solvents. AB - The (13)C NMR spectra of two different iodoalkynes, 1-iodo-1-hexyne (1) and diiodoethyne (2), exhibit a strong solvent dependence. Comparisons of the data with several common empirical models, including Gutmann's Donor numbers, Reichardt's E(N)(T), and Taft and Kamlet's beta and pi, demonstrate that this solvent effect arises from a specific acid-base interaction. Solvent basicity measures such as Donor numbers and beta values correlate well with the alpha carbon chemical shift of 1, but polarity measures such as E(N)(T) and pi do not correlate. The similarity of the solvent effect for 1 and 2 suggests that carbon carbon bond polarization may not play a role in the change in chemical shift, as previously hypothesized. PMID- 14750790 TI - Novel UDP-glycal derivatives as transition state analogue inhibitors of UDP GlcNAc 2-epimerase. AB - The "epimerisation" of UDP-GlcNAc to ManNAc, the first step in the biosynthesis of sialic acids, is catalyzed by UDP-GlcNAc 2-epimerase. In this paper we report the synthesis of transition state based inhibitors of this enzyme. To mimic the assumed first transition state of this reaction (TS 1), we designed and synthesized the novel UDP-exo-glycal derivatives 1-4. We also report herein the synthesis of 5 and 6, the first C-glycosidic derivatives of 2-acetamidoglucal, and the synthesis of the ketosides 7 and 8, which were designed as bis-substrate analogue and bis- product analogue, respectively, to mimic the second step of the reaction via the assumed second transition state TS 2. PMID- 14750791 TI - Lewis acid catalysis in supercritical carbon dioxide. Use of poly(ethylene glycol) derivatives and perfluoroalkylbenzenes as surfactant molecules which enable efficient catalysis in ScCO2. AB - Lewis acid catalysis in supercritical carbon dioxide (CO(2)) was investigated. While solubility of most organic materials is low in scCO(2), poly(ethylene glycol) derivatives or perfluoroalkylbenzenes were found to work as surfactants to dissolve organic materials in scCO(2). In the presence of these molecules, Lewis acid catalyzed organic reactions such as aldol-, Mannich-, and Friedel Crafts-type reactions proceeded smoothly in scCO(2). Formation of emulsions was observed in these reactions, and the systems were studied in detail. PMID- 14750792 TI - Formation of P-ylide under neutral and metal-free conditions: transformation of aziridines and epoxides to conjugated dienes in the presence of phosphine. AB - A general approach to formation of the P-ylide from the reaction of aziridines or epoxides with organophosphine under neutral and metal-free conditions is realized. Conjugated diene derivatives based on this kind of P-ylide were prepared in a facile and convenient way. PMID- 14750793 TI - Metal-catalyzed coupling reactions on an olefin template: the total synthesis of (13E,15E,18Z,20Z)-1-hydroxypentacosa- 13,15,18,20-tetraen-11-yn-4-one 1-acetate. AB - The naturally occurring ant venom (13E,15E,18Z,20Z)-1-hydroxypentacosa 13,15,18,20-tetraen-11-yn-4-one 1-acetate was synthesized stereospecifically using a series of metal-mediated cross-coupling reactions. The use of the difunctional olefin template (E)-1-chloro-2-iodoethylene as the central, pseudosymmetrical building block facilitated a fully convergent and, thus, efficient strategy to prepare this polyunsaturated natural product. PMID- 14750794 TI - Syntheses and activities of new C10 beta-turn peptidomimetics. AB - A program to identify small molecules that mimic or disrupt protein-protein interactions led us to design the peptidomimetics 1-3. Solid-phase syntheses of 1 3 were developed. The purities of the crude materials isolated from the resin tend to be highest for the S- and N-compounds 2 and 3 and better than in the corresponding syntheses of peptidomimetics A. The particular dipeptide units incorporated were chosen to correspond with the turn regions of the neurotrophins (e.g., nerve growth factor [NGF] and the neurotrophin factor-3 [NT-3]). Preliminary studies were performed to access the binding of these analogues to Trk receptors and their ability to induce cell survival (just as NGF and NT-3 do). Several active compounds were identified. However, poor water solubilities of some of the other compounds preclude reliable testing. Consequently, solid phase modifications to the synthetic procedures were investigated to provide access to the derivatives 12-14 in which the aromatic nitro group is replaced by amine, guanidine, or sulfonamide functionalities. The latter are more acceptable pharmacophores than nitro groups and also tend to increase the water solubilities of the peptidomimetics. PMID- 14750795 TI - Intramolecular chiral relay at stereogenic nitrogen. Synthesis and application of a new chiral auxiliary derived from (1R,2S)-norephedrine and acetone. AB - (1R,2S)-Norephedrine has been employed in the synthesis of a novel 3,4,5,6 tetrahydro-2H-1,3,4-oxadiazin-2-one via reductive alkylation with acetone, N nitrosation, reduction, and cyclization. The oxadiazinone was treated with either propionyl chloride or 3-thiophenylpropionyl chloride to afford the corresponding N(3)-acylated oxadiazinones 9a and 9b, respectively. X-ray crystallographic analysis of the N(3)-thiophenylpropionyl oxadiazinone 9b revealed that the C(2) urethane carbonyl and the N(3)-carbonyl are arranged in an anti-periplanar conformation. The oxadiazinones were subsequently applied in the titanium mediated asymmetric aldol addition reaction by treatment with titanium tetrachloride, triethylamine, and a variety of aldehydes at 0 degrees C. The aldol adducts 10a-i and 11a,b were found to have diastereoselectivities ranging from 8:1 to >99:1 favoring the formation of the non-Evans syn configuration. The absolute stereochemistry of the adduct 10a was determined by acid hydrolysis. This process afforded the N(4)-isopropyloxadiazinone 8 and (2S,3S)-3-hydroxy-2 methyl-3-phenylpropanoic acid 14 in >/=95% enantiomeric excess. PMID- 14750796 TI - Spectroscopic correlations between supermolecules and molecules. Anatomy of the ion-modulated electronic properties of the nitrogen donor in monoazacrown-derived intrinsic fluoroionophores. AB - The synthesis, absorption and emission spectra, fluorescence quantum yields, and fluorescence lifetimes of three compound series of trans-4,4'-disubstituted aminostilbenes (1-3) are reported. The chromo-/fluoroionophoric behavior of the monoaza-15-crown-5- (A15C5) and monoaza-18-crown-6 (A18C6)-derived species (1A(5)()-3A(5)() and 1A(6)()-3A(6)()) in acetonitrile and dichloromethane are also investigated. Great similarities in electronic spectroscopic properties (chemical shifts, wavelength, intensity, and lifetime) between the metal ion complexed supermolecules and the corresponding chloro-substituted molecules have been observed: namely, 1A(5)()/Ca(2+)-3A(5)()/Ca(2+) approximately 1A(6)()/Ba(2+) 3A(6)()/Ba(2+) approximately 1C-3C in acetonitrile and 1A(5)()/Na(+) 2A(5)()/Na(+) approximately 1A(6)()/K(+)-2A(6)()/K(+) approximately 1C-2C in dichloromethane. Such spectroscopic correlations allow us to define the metal ion modulated electronic character of the azacrown nitrogen atom in the ground and excited states and, in turn, to gain insights into the observed fluoroionophoric behavior of these probes in terms of the size and direction of fluorescence shifts and intensity variations. PMID- 14750797 TI - Toward the development of a structurally novel class of chiral auxiliaries. Conformational properties of the aldol adducts of oxadiazinones: observation of unusual shielding effects. AB - Asymmetric aldol reactions were conducted with the titanium enolate of N(3) hydrocinnamoyl-3,4,5,6-tetrahydro-2H-1,3,4-oxadiazin-2-one to afford aldol adducts 5a-j. The dominant product of the asymmetric aldol reaction was the non Evans syn adduct as determined by (1)H NMR spectroscopy and X-ray crystallography. When evaluating the (1)H NMR spectra of adducts 5a-j, a highly shielded signal with an average chemical shift of 0.05 ppm was observed. This signal was readily determined to be the C(5)-methyl group of the oxadiazinone. It is presumed that the overall conformation adopted by the aldol adducts in solution places an aromatic ring of the N(3)-substituent in close proximity to the C(5)-methyl group. An investigation of this conformational preference is conducted employing (1)H NMR spectroscopy, X-ray crystallography, and computational methods. PMID- 14750798 TI - The first catalytic asymmetric allylation of imines with the tetraallylsilane TBAF-MeOH system, using the chiral bis-pi-allylpalladium complex. AB - The asymmetric allylation of imines with use of catalytic transition metals with chiral ligands should be a new frontier of the enantioselective C-C bond formation. So far allyltrimethylsilane, allyltrichlorosilane, and allyltrimethoxysilane have been commonly employed with use of either silane activators or dual silane-imine activators. However, tetraallylsilane is untouched in the allylation of aldimines. The first allylation of aldimines with the tetraallylsilane-TBAF-MeOH system with use of the bis-pi-allylpalladium catalyst under catalytic, non-Lewis acid, essentially neutral and very mild reaction conditions has been achieved. The reaction is triggered by dual activation/promotion by TBAF and MeOH in which the fluoride anion activates the C Si bond cleavage and MeOH promotes the facile protonation of intermediate palladium amide. Thus, the synthesis of chiral homoallylamines is achieved in a shorter reaction time and higher yields and enantioselectivities through an efficient, general, and reproducible allylation protocol for imines. PMID- 14750799 TI - General synthesis of unsymmetrical norbornane scaffolds as inducers for hydrogen bond interactions in peptides. AB - Starting from readily accessible endo-cis-(2S,3R)-norbornene dicarboxylic acid benzyl monoester, a general and efficient synthetic approach toward unsymmetrical two-stranded peptidic structures was developed. In these structures the peptide strands are oriented in a parallel geometry. Their synthesis is easily applicable to a variety of amino acids and peptides. Specifically, a norbornane template as molecular scaffold induces hydrogen bonding between the adjacent peptide strands. The specific hydrogen bonding patterns between these strands were revealed by detailed NMR analysis including TOCSY/NOE experiments. PMID- 14750800 TI - Synthesis of DNA with phenanthridinium as an artificial DNA base. AB - A phenanthridinium-containing DNA building block was synthesized as an ethidium nucleoside analogue starting from 3,8-diamino-6-phenyl-phenanthridine. Using this building block, oligonucleotides bearing the phenanthridinium moiety as an artificial DNA base were prepared via automated solid-phase phosphoramidite chemistry. The modified phenanthridinium-containing DNA duplexes were characterized by UV/vis absorption spectroscopy (including the melting behavior), CD spectroscopy, and steady-state fluorescence spectroscopy. These experiments reveal the expected similarity of the synthetic phenanthridinium moiety with noncovalently bound ethidium. More importantly, the results show clearly that the artificial phenanthridinium base is intercalated within the DNA base stack. The counterbase as part of the complementary strand seems to have only a minor influence on the intercalation properties of the phenanthridinium moiety. PMID- 14750801 TI - Total synthesis of the proposed structures of indole alkaloids lyaline and lyadine. AB - The harman-1,4-dihydropyridines 1 and 2, which constitute the originally proposed structures for the indole alkaloids lyaline and lyadine, have been synthesized, and their NMR data have been compared with those available for the natural products. Due to the discrepancies in the spectral data, the structures of lyadine and lyaline should be revised. PMID- 14750802 TI - Characteristics of the two frontier orbital interactions in the Diels-Alder cycloaddition. AB - Two electron-deficient dienes were reacted with a series of twelve electron-poor and electron-rich dienophiles to give, in some cases, the corresponding Diels Alder adducts. Clear differences in the roles played by the two frontier orbital interactions emerged. It was demonstrated that in the case of normal Diels-Alder cycloadditions, the FMO theory could predict the relative reactivities between dienophiles, while in the case of inverse-electron demand Diels-Alder reactions, it could not. It was shown that the dissymmetry in electron-rich dienophiles increases their reactivities. PMID- 14750803 TI - A tin-complexation strategy for use with diverse acylation methods in the preparation of 1,9-diacyldipyrromethanes. AB - The acylation of dipyrromethanes to form 1,9-diacyldipyrromethanes is an essential step in the rational synthesis of porphyrins. Although several methods for acylation are available, purification is difficult because 1,9 diacyldipyrromethanes typically streak extensively upon chromatography and give amorphous powders upon attempted crystallization. A solution to this problem has been achieved by reacting the 1,9-diacyldipyrromethane with Bu(2)SnCl(2) to give the corresponding dibutyl(5,10-dihydrodipyrrinato)tin(IV) complex. The reaction is selective for dipyrromethanes that bear acyl groups at both the 1- and 9 positions but otherwise is quite tolerant of diverse substituents. The diacyldipyrromethane-tin complexes are stable to air and water, are highly soluble in common organic solvents, crystallize readily, and chromatograph without streaking. Four methods (Friedel-Crafts, Grignard, Vilsmeier, benzoxathiolium salt) were examined for the direct 1,9-diacylation of a dipyrromethane or the 9-acylation of a 1-acyldipyrromethane. In each case, treatment of the crude reaction mixture with Bu(2)SnCl(2) and TEA at room temperature enabled facile isolation of multigram quantities of the 1,9 diacyldipyrromethane-tin complex. The diacyldipyrromethane-tin complexes could be decomplexed with TFA in nearly quantitative yield. Alternatively, use of a diacyldipyrromethane-tin complex in a porphyrin-forming reaction (reduction with NaBH(4), acid-catalyzed condensation with a dipyrromethane, DDQ oxidation) afforded the desired free base porphyrin in yield comparable to that obtained from the uncomplexed diacyldipyrromethane. The acylation/tin-complexation strategy has been applied to a bis(dipyrromethane) and a porphyrin dipyrromethane. In summary, the tin-complexation strategy has broad scope, is compatible with diverse acylation methods, and greatly facilitates access to 1,9 diacyldipyrromethanes. PMID- 14750804 TI - Synthetic peptidoglycan substrates for penicillin-binding protein 5 of Gram negative bacteria. AB - The major constituent of the bacterial cell wall, peptidoglycan, is comprised of repeating units of N-acetylglucosamine (NAG) and N-acetylmuramic acid (NAM) with an appended peptide. Penicillin-binding proteins (PBPs) are involved in the final stages of bacterial cell wall assembly. Two activities for PBPs are the cross linking of the cell wall, carried out by dd-transpeptidases, and the dd-peptidase activity, that removes the terminal d-Ala residue from peptidoglycan. The dd peptidase activity moderates the extent of the cell wall cross-linking. There exists a balance between the two activities that is critical for the well-being of bacterial cells. We have cloned and purified PBP5 of Escherichia coli. The membrane anchor of this protein was removed, and the enzyme was obtained as a soluble protein. Two fragments of the polymeric cell wall of Gram-negative bacteria (compounds 5 and 6) were synthesized. These molecules served as substrates for PBP5. The products of the reactions of PBP5 and compounds 5 and 6 were isolated and were shown to be d-Ala and the fragments of the substrates minus the terminal d-Ala. The kinetic parameters for these enzymic reactions were evaluated. PBP5 would appear to have the potential for turnover of as many as 1.4 million peptidoglycan strands within a single doubling time (i.e., generation) of E. coli. PMID- 14750805 TI - Novel enhancement of diastereoselectivity of [2 + 2] photocycloaddition of chiral cyclohexenones to ethylene by adding naphthalenes. AB - The additive effect on the diastereoselective [2 + 2] photocycloaddition of chiral cyclohexenones 1 to ethylene is examined. A novel and fairly efficient method of increasing the diastereoselectivity in the reaction of 1a was elucidated. The de value increased from 56% to 83% by the addition of 1 phenylnaphthalene. The major product 2a was isolated by the recrystallization of the diastereomeric mixture (major/minor = 11/1), of which X-ray analysis confirmed the absolute configuration of the bicyclic system of 2a. Hydrolysis for removing the chiral auxiliary and subsequent esterification afforded the optically pure bicyclo[4.2.0]octanone derivative 5. From the fluorescence spectral analyses and other experimental results, the additive effect is attributed to the complex formation of chiral cyclohexenone 1a and added naphthalenes. PMID- 14750806 TI - Application of a recyclable pseudoephedrine resin in asymmetric alkylations on solid phase. AB - A pseudoephedrine resin has been successfully employed in asymmetric alkylations on solid phase. Immobilized pseudoephedrine amides are conveniently prepared by the one-step attachment of pseudoephedrine to Merrifield resin through the hydroxyl group and subsequent acylation on nitrogen. Deprotonation and alkylation of the resin-bound amides proceeds smoothly. Ketones and alcohols are cleaved from the resin in high enantiomeric excess and moderate to good overall yield. The parallel, asymmetric solid-phase synthesis of a small library of chiral ketones and alcohols has been carried out to illustrate the utility of the approach. Finally, the pseudoephedrine resin can be conveniently recycled and utilized with no significant loss in the yield or enantiomeric excess of the products. PMID- 14750807 TI - Hydrolysis of 2',3'-O-methyleneadenos-5'-yl bis(2',5'-di-O-methylurid-3'-yl) phosphate, a sugar O-alkylated trinucleoside 3',3',5'-monophosphate: implications for the mechanism of large ribozymes. AB - Hydrolytic reactions of 2',3'-O-methyleneadenos-5'-yl bis(2',5'-di-O-methylurid 3'-yl) phosphate (1), a sugar O-alkylated trinucleoside 3',3',5'-monophosphate, have been followed by RP HPLC over a wide pH range. Under neutral and mildly acidic conditions, the only reaction observed was a pH-independent cleavage of the O-C5' bond of the 5'-linked nucleoside. Under more alkaline conditions nucleophilic attack by hydroxide ion starts to compete. The reaction is first order in [OH(-)] and becomes predominant at pH 10. Each of the 3'-linked nucleosides is displaced 2.9 times as readily as the 5'-linked one. To determine the beta(lg) value for the hydroxide ion catalyzed hydrolysis of 1, two diesters (2a,b) having 2',3'-O-methyleneadenosine (7) and 2',5'-di-O-methyluridine (4) as leaving groups were hydrolyzed under alkaline conditions. Since the beta(lg) value for this reaction is known, DeltapK(a) between 4 and 7 could be calculated. The beta(lg) for the hydrolysis of 1 was estimated to be -0.5 with use of this information. The mechanisms of the partial reactions and the role of leaving group properties in ribozyme reactions of large ribozymes are discussed. PMID- 14750808 TI - Facile syntheses of oxazolines and thiazolines with N-acylbenzotriazoles under microwave irradiation. AB - Microwave reactions of 2-amino-2-methyl-1-propanol (2) or 2-aminoethanethiol hydrochloride (4) with readily available N-acylbenzotriazoles 1a-j in the presence of SOCl(2) produced 2-substituted 2-oxazolines 3a-j in 84-98% yields and 2-substituted thiazolines 5a-i in 85-97% yields, respectively. With use of this method chiral oxazoline 6, bisoxazoline 7, bisthiazoline 8, and 5,6-dihydro-4H 1,3-oxazines 9 or 10 have also been prepared in 82-96% yields. These results demonstrate a new application of N-acylbenzotriazoles in the preparation of oxazolines and thiazolines under mild conditions and short reaction times with microwave irradiation. PMID- 14750809 TI - Synthesis of gamma,delta-unsaturated and delta,epsilon-unsaturated alpha-amino acids from fragmentation of gamma- and delta-lactones. AB - A noncoded amino acid of cyclomarin A (1) was synthesized in a racemic fashion. The method employs a six-membered ring template to control the relative stereochemistry and introduction of the functional groups. Ultimately, Pd catalyzed fragmentation of the lactone provided gamma,delta-unsaturated and delta,epsilon-unsaturated alpha-amino acids. A Pd-catalyzed ring opening of a gamma-lactone is also reported. PMID- 14750810 TI - Conformational studies by dynamic NMR. 98. Stereodynamics of bond rotation in beta-hydroxyesters. AB - The dynamic processes due to restricted intramolecular motions have been observed in a number of variously substituted beta-hydroxyesters by variable-temperature NMR spectroscopy. The barriers for three such processes (i.e. Ph-C, t-Bu-C, and C(alpha)-C(beta) bond rotation) have been measured. PMID- 14750811 TI - Straightforward asymmetric entry to highly functionalized 3-substituted 3-hydroxy beta-lactams via Baylis-Hillman or bromoallylation reactions. AB - The reaction of various activated vinyl systems, including 2-cyclopenten-1-one, with enantiopure azetidine-2,3-diones 1 was promoted by DABCO to afford the corresponding optically pure Baylis-Hillman adducts 2 without detectable epimerization. However, the reaction of alpha-keto lactams 1 with but-3-yn-2-one was not as successful, giving the corresponding beta-halo Baylis-Hillman adduct in low yield. Metal-mediated bromoallylation reaction between 2,3-dibromopropene and azetidine-2,3-diones 1 was investigated in aqueous media. Surprisingly, indium was unable to promote the bromoallylation reaction of alpha-keto lactams 1, but the Sn-Hf(4)Cl-promoted bromoallylation of ketones 1 proceeded efficiently to achieve bromohomoallyl alcohols 5 as single diastereomers. On this basis, simple and fast protocols for the asymmetric synthesis of the potentially bioactive 3-substituted 3-hydroxy-beta-lactam moiety were developed. PMID- 14750812 TI - Novel synthesis of highly functionalized 14-beta-hydroxysteroids related to batrachotoxin and ouabain. AB - The use of anionic polycyclization was investigated in an effort to develop a versatile and convergent synthesis of advanced tetracyclic intermediates of batrachotoxin and ouabain analogues. Two new 5-(trialkylsilyl)-2-cyclohexenones as A ring precursors and a new Nazarov intermediate (D ring precursor) were prepared for this purpose. The reaction of the unsaturated beta-keto aldehyde A ring precursor with the enolate of the Nazarov intermediate afforded, after subsequent transformations, a 14-beta-hydroxysteroid with complete control of stereochemistry. PMID- 14750813 TI - Facile and effective copper-mediated cyclization reaction of cyclopropylideneacetic acids (or esters) and cyclopropylideneacetonitriles. AB - The full details of the copper-mediated cyclization reaction of cyclopropylideneacetic acids (or esters) and cyclopropylidenenitriles, the synthetic application of this reaction, and the study of the reaction mechanism are reported. PMID- 14750815 TI - Transition structures, energetics, and nucleus-independent chemical shifts for divinylcyclobutene-to-cyclooctatriene rearrangement: a DFT study. AB - The minimum energy reaction paths and nucleus-independent chemical shifts (NICS) for the Cope rearrangement of cis-3,4-divinylcyclobutene, obtained by (U)B3LYP/6 31G calculations, are reported. Three transition structures (endo-boatlike, chairlike, and exo-boatlike) have been located, giving rise to formation of cis,cis,cis-, cis,cis,trans-, and trans,cis,trans-1,3,5-cyclooctatrienes, respectively. The minimum energy path proceeds through an endo-boatlike, aromatic transition structure. The reaction path of the rearrangement is intervened by enantiomerization saddle point of the product. NICS values calculated for transition structures agree qualitatively with their activation energy and reaction exothermicity orders. Cope rearrangement and electrocyclic ring-opening processes of cis-3,4-divinylcyclobutene are competitive, but the former is relatively more favored and exothermic than the latter. PMID- 14750814 TI - Synthesis of 4-amino-5-H-2,3-dihydroisothiazole-1,1-dioxide ring systems on sugar templates via carbanion-mediated sulfonamide intramolecular cyclization reactions (CSIC protocols) of glyco-alpha-sulfonamidonitriles. AB - The carbanion-mediated sulfonate intramolecular cyclizations (CSIC protocols) of glyco-alpha-sulfonamidonitriles derived from readily available monosaccharides have been extensively investigated using potassium carbonate, cesium carbonate, n BuLi, and LDA as bases. As a result, a series of enantiomerically pure spiro(4 amino-5-H-2,3-dihydroisothiazole-1,1-dioxide) derivatives have been prepared efficiently and isolated in good yield. The synthesis of these new bicyclic systems is key to accessing a novel range of aza analogues of TSAO nucleosides (ATSAOs). PMID- 14750816 TI - A silicon tether approach for addition of functionalized radicals to chiral alpha hydroxyhydrazones: diastereoselective additions of hydroxymethyl and vinyl synthons. AB - Stereocontrolled additions of hydroxymethyl and vinyl groups to chiral alpha hydroxyhydrazones can be achieved by radical cyclizations using bromomethyl or vinyl radical precursors tethered via a temporary silicon connection. Tin mediated 5-exo radical cyclization of alpha-hydroxyhydrazones using a silicon tethered bromomethyl group, followed by oxidative removal of the tether, provides anti-2-hydrazino 1,3-diols in good yield. Tandem thiyl radical addition cyclization of alpha-hydroxyhydrazones using a silicon-tethered vinyl group, followed by treatment with potassium fluoride, affords acyclic allylic anti hydrazino alcohols in good yield. The thiyl addition-cyclization method has been successfully extended to the use of alpha,beta-dihydroxyhydrazones without prior protection or hydroxyl differentiation. Diastereoselection in both reaction types increases with increasing A values of the appended groups, consistent with prediction by the Beckwith-Houk model for stereocontrol in 5-hexenyl radical cyclizations. PMID- 14750817 TI - Alpha-nitro ketone as an electrophile and nucleophile: synthesis of 3-substituted 2-nitromethylenetetrahydrothiophene and -tetrahydrofuran as drosophila nicotinic receptor probes. AB - 3-(6-Chloropyridin-3-yl)methyl-2-nitromethylenetetrahydrothiophene 2 and tetrahydrofuran 3 were synthesized through novel approaches using alpha-nitro ketone intermediates as an electrophile and nucleophile, respectively. The 2 nitromethylenetetrahydrothiophene 2 was formed exclusively as the Z-isomer through intramolecular attack by a thiol substituent at the carbonyl group of an alpha-nitro ketone, in which the alpha-nitro ketone served as an electrophile. In contrast, the corresponding 2-nitromethylenetetrahydrofuran 3, not accessible by the above route due to limited stability, was prepared as a mixture of E- and Z isomers by intramolecular attack of the alpha-nitro ketone enol anion in which the deprotonated alpha-nitro ketone served as a nucleophile. These compounds, together with the corresponding 2-nitromethylenepyrrolidine (1), were used to probe the Drosophila neonicotinoid-nicotinic acetylcholine receptor interaction. PMID- 14750818 TI - Simultaneously electrogenerated cycloaddition partners for regiospecific inverse electron-demand Diels-Alder reactions: a route for polyfunctionalized 1,4 benzoxazine derivatives. AB - A multistep one-pot electrochemical synthesis of a variety of complex 2 alkylamino-1,4-benzoxazine derivatives is described. The reactions are regiospecific and diastereospecific in the case of heterocyclic annulation. This cascade sequence, wherein both cycloaddition partners are generated in situ, at room temperature, under metal-free conditions, allows the inverse-electron-demand Diels-Alder reaction of an o-iminoquinone diene and a secondary alkylenamine dienophile, two chemically nonaccessible unstable entities. To increase the molecular diversity, a variant in which the enamine is separately prepared completes the aforementioned procedure. The extension of this reaction should be useful to generate libraries of heterocycles. PMID- 14750819 TI - Total synthesis of the boron-containing ion carrier antibiotic macrodiolide tartrolon B. AB - The first total synthesis of the boron-containing macrodiolide antibiotic tartrolon B is reported in full detail. Two convergent approaches to the target compound are described, the first of which eventually failed, due to sensitive functionality. In the second, successful route the key step was a stereoselective boron-mediated aldol addition of a bicyclic acetonide protected ketone to a diene aldehyde. In this case the synthesis could be completed without major problems, using a Yamaguchi dimerization macrolactonization endgame. PMID- 14750820 TI - Recognition through self-assembly. A quadruply-hydrogen-bonded, strapped porphyrin cleft that binds dipyridyl molecules and a [2]rotaxane. AB - Quadruply-hydrogen-bonded porphyrin homodimer Zn1.Zn1 has been designed, assembled, and evaluated as a supramolecular cleft-featured receptor for its ability to bind dipyridyl guests in chloroform-d. Monomer Zn1 consists of a 2 ureidopyrimidin-4(1H)-one unit, which was initially reported by Meijer et al., and a zinc porphyrin unit. The zinc porphyrin is strapped with an additional aliphatic chain for controlling the atropisomerization of porphyrin. The 2 ureidopyrimidin-4(1H)-one unit dimerizes exclusively in chloroform even at the dilute concentration of 10(-)(4) M, while the two "strapped" zinc porphyrin units of the homodimer provide additional binding sites for selective guest recognition. (1)H NMR studies indicate that the new homodimer Zn1.Zn1 adopts an S type conformation due to strong donor-acceptor interaction between the electron rich porphyrin units and the electron-deficient 2-ureidopyrimidin-4(1H)-one unit. (1)H NMR, UV-vis, and vapor pressure osmometry investigations reveal that Zn1.Zn1 could function as a new generation of assembled supramolecular cleft, to be able to not only efficiently bind linear dipyridyl molecules 14-17, resulting in the formation of stable termolecular complexes, with K(aasoc) values ranging from 3.8 x 10(6) to 8.9 x 10(7) M(-)(1), but also strongly complex a hydrogen-bond assembled [2]rotaxane, 18, which consists of a rigid fumaramide thread and a pyridine-incorporated tetraamide cyclophane, with K(aasoc) = 1.2 x 10(4) M(-)(1). (1)H NMR competition experiments reveal that complexation to the dipyriyl guests also promotes the stability of the quadruply-hydrogen-bonded dimeric receptor. PMID- 14750821 TI - Pd(0)-catalyzed conjugate addition of benzylzinc chlorides to alpha,beta-enones in an atmosphere of carbon monoxide: preparation of 1,4-diketones. AB - Pd(0)-catalyzed conjugate addition of benzylzinc chloride to methyl vinyl ketone in the presence of chlorotrimethylsilane and lithium chloride in an atmosphere of carbon monoxide at room temperature afforded 1-phenyl-2,5-hexanedione monosilyl enol ether. In this catalytic carbonylation, four components are connected in one reaction. Successive acidic workup generated a variety of 1,4-diketones from substituted benzylzinc chlorides or related compounds and alpha,beta-enones. Some products were converted to cyclopentenones or five-membered heterocyclic compounds containing an N, O, or S atom. PMID- 14750822 TI - Magnesium-induced copper-catalyzed synthesis of unsymmetrical diaryl chalcogenide compounds from aryl iodide via cleavage of the Se-Se or S-S bond. AB - The methodology for a copper-catalyzed preparation of diaryl chalcogenide compounds from aryl iodides and diphenyl dichalcogenide molecules is reported. Unsymmetrical diaryl sulfide or diaryl selenide can be synthesized from aryl iodide and PhYYPh (Y = S, Se) with a copper catalyst (CuI or Cu(2)O) and magnesium metal in one pot. This reaction can be carried out under neutral conditions according to an addition of magnesium metal as the reductive reagent. Furthermore, it is efficiently available for two monophenylchalcogenide groups generated from diphenyl dichalcogenide. PMID- 14750823 TI - Synthesis and functional properties of strongly luminescent diphenylamino end capped oligophenylenes. AB - Two novel homologous series of oligophenylenes (OPPs) symmetrically end-capped with diphenylamino groups and asymmetrically end-capped with anthryldiphenylamino groups were successfully synthesized by a convergent approach with use of palladium-catalyzed homo- and cross-coupling of arylboronic acids. The absorption maxima of both diphenylamino end-capped OPP series do not vary with the chain length although the molar absorptivities increase sequentially. On the other hand, the emission maxima slightly shift to longer wavelengths when the phenylene unit increases in the series. All the diphenylamino end-capped oligomers exhibit very large fluorescence quantum yields (81-89%). They also exhibit low first ionization potentials, corresponding to the oxidation of the triarylamino moiety, which are essentially unaffected by the oligomeric length extension. The good thermal stabilities of these oligomers allowed the fabrication of multilayer light-emitting devices and their investigations. PMID- 14750824 TI - Hammett correlations in the photosensitized oxidation of 4-substituted thioanisoles. AB - Singlet oxygen is quenched by a series of 4-substituted thioanisoles (methoxy to nitro), with rate constant k(t) = 7 x 10(4) to 7 x 10(6) M(-)(1) s(-)(1), close to the value observed for the myoglobin-catalyzed sulfoxidation of the same sulfides. Correlations with sigma (rho = -1.97) and with E(ox) (slope -3.9 V( )(1)) are evidence for an electrophilic mechanism. In methanol sulfoxides are formed (85%) via an intermediate quenched by diphenyl sulfoxide; competing minor paths lead to arylthiols, arylsulfenic acid, and aryl sulfoxides. In aprotic solvents, the sulfoxidation is quite sluggish, but carboxylic acids (mostly 100. The protonated persulfoxide is formed in this case and acts as an electrophile with sulfides, again with a rate constant correlating with sigma (rho = -1.78). PMID- 14750825 TI - Convenient processes for the synthesis of aromatic ketones from aryl bromides and carboxylic anhydrides using a cobalt catalysis. AB - The cross-coupling of various para- and meta-substituted aromatic bromides, mostly bearing sensitive moieties, with several carboxylic acid anhydrides is reported. This reaction can be carried out in two steps, by forming an aromatic organozinc reagent via cobalt catalysis in the first step, or even more interestingly in a single step, also by using a cobalt-based catalyst. The aromatic ketones are obtained by these new, mild, and convenient methods in 30 79% yields versus starting aryl bromide. Results are also disclosed that suggest the role played by cobalt species in the coupling of organozinc reagents with electrophiles could be similar to those of more commonplace transition metal complexes. PMID- 14750826 TI - Efficient fluoride-selective fluorescent host: experiment and theory. AB - A new naphthalene derivative containing a urea group at the 1,8-position of naphthalene was synthesized and showed a unique absorption and fluorescence peak with fluoride ion. Calculations suggested that a new peak was attributed to the increased anionic character of urea nitrogen due to the strongly charged hydrogen bonding between fluoride and amide protons of the urea. The fluoride selectivity among halides (F(-), Cl(-), Br(-)) comes from the fact that the fluoride approaches much closer to the amide protons than other halides and resides in the cavity with fast dynamics. The nature of electronic transitions that were analyzed from the calculations by the collective electronic oscillator method also supports the anionic nature of the complex between host and fluoride. PMID- 14750827 TI - Impact of the solvent on the conformational isomerism of calix[4]arenes: a study based on continuum solvation models. AB - The influence of solvation on the conformational isomerism of calix[4]arene and p tert-butylcalix[4]arene has been investigated by using the continuum model reported by Miertus, Scrocco, and Tomasi (MST). The quantum mechanical (QM) and semiclassical (SC) formalisms of the MST model have been considered for two different solvents (chloroform and water). The suitability of the QM-MST and SC MST methods has been examined by comparison with previous results derived from classical molecular dynamics (MD) simulations with explicit solvent molecules. The application of the continuum model to the solute configurations generated by using in vacuo classical MD simulations provides a fast strategy to evaluate the effects of the solvent on the conformational preferences of calixarenes. These encouraging results allow us to propose the use of continuum models to solutes with complex molecular structures, which are traditionally studied by MD simulations. PMID- 14750828 TI - Diastereoselective dl-hydrocoupling of benzalacetones by electroreduction. AB - Electroreduction of benzalacetones with an undivided cell in Et(4)NOTs/acetonitrile gave cyclized dl-hydrodimers as mixtures of two diastereomers. The hydrodimerization proceeded stereoselectively to afford linear dl-hydrodimers, and the following cyclization led to two thermodynamically stable diasteromers of cyclopentanols. PMID- 14750829 TI - Flexible bidentate pyridine and chiral ligands in the self-assembly of supramolecular 3-D cages. AB - Discrete, nanoscopic 3-D cages are prepared in high yield via coordination-driven self-assembly from a variety of building blocks, including bidentate 3 substituted pyridines, chiral, and silicon-based tripods. All are characterized by NMR ((31)P, (1)H) and electrospray ionization mass spectrometry. PMID- 14750830 TI - Novel route to functionalized cyclooctanoids via [5+3] cycloaddition. AB - The self-dimerization of 3-oxidopyrylium leading to stereocontrolled formation of highly functionalized cyclooctanoids is described. Different functionalities were introduced on the dimer (3) and the stereochemical outcome was determined by single-crystal X-ray analysis. It is noteworthy that the hydrogenation of 3 in ethanol solvent gave the transannulated product 5, whereas the expected dihydro product 4 was obtained when the reaction was run in nonnucleophilic solvents. The mechanistic pathway is discussed. PMID- 14750831 TI - An enantiomerically pure 1,5,7-trimethyl-3-azabicyclo[3.3.1]nonan- 2-one as 1H NMR shift reagent for the ee determination of chiral lactams, quinolones, and oxazolidinones. AB - The chiral lactam 1 (or its enantiomer ent-1) was shown to be an effective (1)H NMR shift reagent for the ee determination of chiral lactams, quinolones, and oxazolidinones. It was successfully employed in many cases in which a detection of enantiomers by chromatographic methods failed. The method was extended to a broader range of simple substrates bearing a lactam moiety to evaluate its scope. The NH signals of the substrate enantiomers showed the strongest separation and were used for (1)H NMR integration. In most cases, compound 1 (1.5 equiv; 0.06 M solution) induced a baseline separation of the NH signals and it can consequently be regarded as a generally applicable shift reagent for chiral products with a lactam moiety. PMID- 14750832 TI - Reactions of a carbamoylstannane with acid chlorides: highly efficient synthesis of alpha-oxo amides. AB - Treatment of acid chlorides with a carbamoylstannane under mild conditions (mostly rt for a few hours) affords alpha-oxo amides in high yields. Vicinal polycarbonyl compounds are also obtained, although spontaneous decarbonylation occasionally occurs. PMID- 14750833 TI - A new strategy for the construction of the imidazo[1,5-a]quinoxalin-4-one ring system and its application to the efficient synthesis of BMS-238497, a novel and potent Lck inhibitor. AB - A new efficient strategy was developed for the construction of the imidazo[1,5 a]quinoxalin-4-one ring system. The new method involves condensation of o nitroaniline with glyoxylate in methanol followed by treatment of the resulting alpha-(o-nitroanilino)-alpha-methoxy acetate with tosylmethyl isocyanide (TosMIC) reagent to give 1-(o-nitrophenyl)imidazole-5-carboxylate. Reductive cyclization of the nitro imidazole carboxylate afforded imidazo[1,5-a]quinoxalin-4-one in three steps and 60% overall yield. The new method was successfully applied to the synthesis of BMS-238497, a novel and potent Lck inhibitor. PMID- 14750834 TI - Asymmetric synthesis of dialkyloxy-3-alkylammonium cationic lipids. AB - The cationic diether-linked cytofectin DOTMA (available commercially as a mixture, Lipofectin comprised of DOTMA:DOPE, 1:1) and analogues including DIMRIE and DORIE are frequently used for in vitro and in vivo transfections. Despite this wide usage direct synthetic routes to the optical isomers have received little attention to date. Here we describe strategies to synthesize enantiomers of DOTMA and analogues, including an extremely concise procedure to the trimethylammonium salts. One strategy utilized N-protection, as the imine, with concomitant ether formation and deprotection during the workup. Methylation of the 1-amino-2,3-dialkyloxypropane then generated the trimethylammonium cationic lipids directly. This methodology was extended to synthesize a novel headgroup functionalized lipid. A second route was also developed using an alternative chiral synthon. PMID- 14750835 TI - Efficient synthesis of exo-olefinated deoxoartemisinin derivatives by Ramberg Backlund rearrangement. AB - 10-exo-Bromoalkylidene- and benzylidenedeoxoartemisinins were synthesized from corresponding 10-alkanesulfonyldihydroartemisinin and 10 phenylmethanesulfonyldihydroartemisinin using a highly efficient, mild, and simple Ramberg-Backlund rearrangement. PMID- 14750836 TI - Synthesis of alkylated aminofluorenes by palladium-catalyzed substitution at halofluorenes. AB - New N-substituted 2-amino-9,9-dialkylfluorenes optionally bearing electron withdrawing substituents such as nitro or cyano in position 7 can be synthesized starting from 2-halo-9,9-dialkylfluorenes by Pd-catalyzed substitution with amines. Chiral amino groups can be introduced by this method too. 2-N,N Dimethylamino-7-nitro-9H-fluorene was obtained in a convenient way by reductive amination. The N-substituted 2-amino-7-nitro-9H-fluorenes are promising candidates for fluorescence probes for femtosecond solvation dynamics. PMID- 14750837 TI - Nucleophilic partners in the tandem conjugate addition-Dieckmann condensation reaction: 1. synthesis of 1,2,3-trisubstituted naphthalenes. AB - The scope and limitations of the tandem conjugate addition-Dieckmann condensation for the construction of 1,2,3-trisubstituted naphthalenes is defined. Viable nucleophilic partners in this methodology include organocuprates, active methylenes, and a variety of heteroatom initiators. PMID- 14750838 TI - Synthesis of optically pure highly functionalized gamma-lactams via 2-azetidinone tethered iminophosphoranes. AB - A synthesis of optically pure densely functionalized gamma-lactams starting from 2-azetidinone-tethered iminophosphoranes has been developed. Full chirality transfer has been accomplished from the enantiomerically pure 2-azetidinones. The addition of lithium acetylides to 4-oxoazetidine-2-carbaldehydes at -78 degrees C smoothly yielded propargylic alcohols with excellent diastereoselectivities. Propargylic alcohols were converted to mesylates, which by exposure to sodium azide afforded the corresponding azides. Treatment of beta-lactams bearing an azido side chain with triphenylphosphine (TPP) gave lambda(5)-phosphazenes (iminophosphoranes, phosphine imines), which were not isolated. The sodium methoxide promoted reaction of the phosphazene beta-lactams smoothly provided gamma-lactams, through a N1-C2 bond breakage process on the four-membered lactam with concomitant ring expansion, followed by hydrolysis. PMID- 14750839 TI - A direct retro-Reformatsky fragmentation: formal ring enlargement of cyclic ketones for novel and practical synthesis of heterocyclic enamines. AB - A novel and practical synthesis of heterocyclic enamines has been developed from the formal ring enlargement of cyclic ketones, which comprised the retro Reformatsky fragmentation reaction as a key step. Under alkaline bromination conditions, the Reformatsky adducts derived from five- to seven-membered cyclic ketones underwent efficiently a direct retro-Reformatsky fragmentation, followed by spontaneous alpha,alpha-dibromination, to produce alpha,alpha,omega-tribromo beta-ketoester compounds in a one-pot reaction. Highly regioselective reduction of alpha,alpha,omega-tribromo-beta-ketoesters with Cu-Zn alloy under mild conditions afforded omega-bromo-beta-ketoesters in good to excellent yields. Treatment of omega-bromo-beta-ketoesters with sodium azide followed by intramolecular aza-Wittig reaction or catalytic hydrogenation furnished heterocyclic secondary enamines, while a straightforward cyclocondensation of omega-bromo-beta-ketoesters with amines led to the formation of heterocyclic tertiary enamines. PMID- 14750840 TI - Carbon composite microelectrodes: charge percolation and electroanalytical performance. AB - Microelectrodes based on two different epoxy-graphite composites (Araldite M/HY5162 and Araldite-PY302-2/HY943) that are compatible with organic solvents have been developed and characterized. The variation in the bulk conductivity with graphite particle loading is described by percolation theory and indicates that the particles interact strongly with one another. The percolation threshold is 52% v/v loading of graphite, and this composite exhibits a bulk conductivity of 15 S m(-1). Microdisk electrodes of 25-microm diameter were produced by first etching a microcavity at the tip of a platinum microelectrode, which was then packed with a composite containing 60% v/v graphite so as to optimize both electrical conductivity and the electrode stability in acetonitrile and methanol solutions. Solution phase voltammetry of ferrocene is nearly ideal, and the responses are dominated by radial diffusion (slow scan rates) and semi-infinite linear diffusion (fast scan rates). The microelectrodes display high signal-to noise ratios, good sensitivity, and low detection limits. The response times given by the product of the resistance, R, and capacitance, C, are 7.5 x 10(-4) and 1.4 x 10(-1) s for the Araldite M and PY302-2 composites, respectively. Although these response times are significantly slower than those associated with microelectrodes based on carbon fibers or metal wires, they are sufficient for time-resolved electroanalytical applications. The long response times arise from the large composite resistances, 3.1 x 10(11) and 8.3 x 10(11) Omega cm(-2) for Araldite M and PY302-2, respectively. Voltammetry of ferrocene in the absence of deliberately added supporting electrolyte is also reported. Significantly, indistinguishable slopes and intercepts for a calibration curve of peak current vs ferrocene concentration where 2 < [ferrocene] < 50 microM are obtained in the presence and absence of supporting electrolyte. PMID- 14750841 TI - Preparation, characterization, and time-resolved fluorometric application of silica-coated terbium(III) fluorescent nanoparticles. AB - Novel silica-coated terbium(III) chelate fluorescent nanoparticles have been prepared and characterized as a new type of fluorescence probe for highly sensitive time-resolved fluorescence bioassay. The preparation was carried out in a water-in-oil microemulsion containing a strongly fluorescent Tb(3+) chelate, N,N,N(1),N(1)-[2,6-bis(3'-aminomethyl-1'-pyrazolyl) phenylpyridine]tetrakis(acetate)-Tb(3+), Triton X-100, hexanol, and cyclohexane by controlling hydrolysis of tetraethyl orthosilicate. The nanoparticles are spherical and uniform in size, 42 +/- 3 nm in diameter, strongly fluorescent, and highly photostable and have enough of a long fluorescence lifetime (1.52 ms) for time-resolved fluorescence measurement. A stable and nontoxic method was developed for the surface modification and protein immobilization of the nanoparticles. As a model of application, the nanoparticle-labeled streptavidin was prepared and used in a sandwich-type time-resolved fluoroimmunoassay of human prostate-specific antigen (PSA) by using a 96-well microtiter plate as the solid phase carrier. The method gives a detection limit of 7.0 pg/mL for the PSA assay. PMID- 14750842 TI - A microphysiometer for simultaneous measurement of changes in extracellular glucose, lactate, oxygen, and acidification rate. AB - A microphysiometer capable of measuring changes in extracellular glucose, lactate, oxygen, and acidification rate has been developed by incorporating modified electrodes into a standard Cytosensor Microphysiometer plunger. Glucose and lactate are measured indirectly at platinum electrodes by amperometric oxidation of hydrogen peroxide, which is produced from catalysis of glucose and lactate at films containing their respective entrapped oxidase. Oxygen is measured amperometrically at a platinum electrode coated with a Nafion film, while the acidification rate is measured potentiometrically by a Cytosensor Microphysiometer. Analytical information is obtained during the Cytosensor stop flow cycles, where the electrodes measure changes in the extracellular medium corresponding to the consumption or production of the analyte by the cells. Modification of the Cytosensor plunger for multianalyte determination is described, and the operation of the technique is illustrated by the simultaneous measurement of all four analytes during the addition of fluoride and DNP to Chinese hamster ovary cells and fluoride and antimycin A to mouse fibroblast cells. Cell metabolic recovery and dynamics after exposure to agents can also be observed in specific cases. PMID- 14750843 TI - Determination of primary and secondary standards and characterization of appropriate salt bridges for pH measurements in formamide. AB - For the first time, the standardization for pH measurements is here implemented in the domain of superpermittive media. The nonaqueous solvent studied is formamide (epsilon = 109.5 at 298.15 K) for which three primary standards and two secondary standards have been determined, whose excellent internal consistency has also been demonstrated by specific cell measurements. A comparison of the pH scale in formamide with the aqueous scale has been duly tried by accounting for the primary medium effect on the H+ ion. Furthermore, as a result of an ad hoc supplementary systematic investigation, three salt bridges of appropriate level of equitransference in formamide, that is, NH(4)Cl, NH(4)Br, and NH(4)I, have been characterized for abating the liquid junction potentials intervening in the pH-measuring cell to enable the user to carry out regular pH measurements and related controls. PMID- 14750844 TI - Improved planar amperometric nitric oxide sensor based on platinized platinum anode. 1. Experimental results and theory when applied for monitoring NO release from diazeniumdiolate-doped polymeric films. AB - An improved miniature amperometric nitric oxide sensor design with a planar sensing tip (ranging from 150 microm to 2 mm in diameter) is reported. The sensor is fabricated using a platinized platinum anode and a Ag/AgCl cathode housed behind a microporous poly(tetrafluoroethylene) (PTFE; Gore-tex) gas-permeable membrane. Platinization of the working platinum electrode surface dramatically improves the analytical performance of the sensor by providing approximately 10 fold higher sensitivity (0.8-1.3 pA/nM), approximately 10-fold lower detection limit (< or =1 nM), and extended (at least 3-fold) stability (>3 d) compared to sensors prepared with bare Pt electrodes. These improvements in performance arise from increasing the kinetics and lowering the required potential for the 3 electron oxidation of NO to nitrate, relative to that observed using a nonplatinized working electrode. The outer porous PTFE membrane provides complete selectivity for NO over nitrite ions (up to 10 mM nitrite). The new sensor is applied for surface measurements of NO released from diazeniumdiolate-loaded silicone rubber films (SR-DACA-6/N(2)O(2)). The effects of sensor size (for sensor dimensions of 0.15-, 1-, and 2-mm o.d.) and the distance of the sensor from the surface of the NO-emitting polymer film are investigated via experiments as well as theoretical calculations. A significant analyte trapping effect is demonstrated, the degree of which depends on the sensor size and its distance from the surface. It is further demonstrated that surface NO concentrations for fresh SR-DACA-6/N(2)O(2) loaded films are also influenced by the polymer film thickness, with thicker films generating higher surface concentrations of NO. PMID- 14750845 TI - Improved planar amperometric nitric oxide sensor based on platinized platinum anode. 2. Direct real-time measurement of NO generated from porcine kidney slices in the presence of l-arginine, l-arginine polymers, and protamine. AB - Nitric oxide generation from porcine kidney slices is assessed using a new planar NO-selective amperometric sensor. The planar shape of the sensor allows for direct NO measurements near the surface (10 microm) of renal tissue slices in real time. Renal NO production may be modulated by the addition of L-arginine, arginine homopolymers (R2, R6, R10), and protamine, all of which can potentially transport across cellular membranes and provide a substrate for nitric oxide synthase within kidney parenchyma. Real-time amperometric measurements demonstrate that most L-arginine species can translocate across the cell membrane and rapidly increase NO production. However, no increase in NO generation is observed when the dimer of L-arginine (R2) is added to the solution bathing the tissue, suggesting that this species cannot permeate cell membranes. The degree of enhancement in NO generation observed for L-arginine and the larger peptides depends on the structure and follows the following sequence: R10 (decamer) > protamine > R6 (hexamer) > L-arginine. Protamine and the R10 decamer, especially, induce the largest increases in NO generation owing to their apparent rapid translocation into cells and subsequent cleavage by proteases to create high intracellular levels of L-arginine. The effect of sensor size (for sensor dimensions of 0.15- and 1-mm outer diameters) on the measured surface NO levels is also examined. The larger sensor traps more NO but hinders access of the L arginine species to the tissue area between the flat distal plane of the sensor and the surface of the kidney slice. The use of such NO-generating peptides may be important in numerous biological systems that depend on NO production, such as ischemia-reperfusion injury and thrombogenesis. PMID- 14750846 TI - Correlation of analyte structures with biosensor responses using the detection of phenolic estrogens as a model. AB - The apparent increase in hormone-induced cancers and disorders of the reproductive tract in wildlife and humans has led to a search for an accurate and reliable method for monitoring endocrine-disrupting chemicals (EDCs). This study presents a generic approach that may allow researchers to establish screening procedures for potential EDCs by correlating the analyte structures with biosensor responses and explain possible reaction mechanisms. A simple amperometric tyrosinase-based biosensor (Tyr-CPE) has been developed for the detection of phenolic EDCs. The investigation of the enzymatic oxidation of selected phenolic estrogens was first carried out using UV-vis spectroscopy. The result was used to correlate sensor responses to enzymatic activity. Natural phytoestrogen polyphenols, including resveratrol (RES), genistein (GEN), and quercetin (QRC), were compared with synthetic estrogens, for example, bisphenol A (BPhA), nonylphenol (NPh), and diethylstilbestrol (DES). The Tyr-CPE biosensor resulted in rapid, simple, and accurate measurement of phenolic estrogens with varying degrees of sensitivity, selectivity, and response times. The sensor responses have been evaluated for the detection of binary and tertiary mixtures of EDCs and natural estrogens. The results showed that BPhA could be successfully discriminated in a composite mixture containing NPh and DES at various ratios. In the case of natural phenolic estrogens GEN, RES, and QRC, the sensor allows the determination of a total phenolic content. The sensor was also validated for the detection of BPhA in a real environmental water sample, and the results was compared with standard ASTM method 9065. Mechanistically, our results indicated that the number of OH groups, the nature and the position of aryl ring substituents, or both could affect the detection limit and the biosensor sensitivity. PMID- 14750847 TI - Surface plasmon resonance spectroscopy based on evanescent field treatment. AB - The reflectance in a surface plasmon resonance (SPR) curve can be expressed in terms of the integration of the product between the evanescent electric field and the imaginary part of the dielectric constant of all absorbing media. The evanescent field in the metal film consists of two fields, one originating at the prism/metal interface and the other at the metal/dielectric interface. Near the resonance angle, the evanescent field strength at the metal/dielectric interface is much greater than that at the prism/metal interface. The evanescent field in dielectric medium has a single origin at the metal/dielectric interface. Due to the optical enhancement at the interface, the amplitude of the evanescent electric field in the dielectric medium is much greater than that in the metal film. This field, however, is not being utilized in conventional SPR where changes in the refractive index of the nonabsorbing dielectric media are of interest. In a system with an absorbing dielectric medium, the absorption of the medium is enhanced by the strong evanescent electric field. The evanescent field distributions in the metal film and in the dielectric medium are significantly altered by the absorbing dielectric, which results in shifting of the resonance angle, increasing of the reflectance, and broadening of the SPR curve. Since the absorption contribution from the absorbing dielectric can be separated from that of the metal film via knowledge of evanescent field distribution, an in-depth analysis of the SPR curve of an absorbing medium and its relationship with the material characteristics are possible. PMID- 14750848 TI - Submicrometric lipobead-based fluorescence sensors for chloride ion measurements in aqueous solution. AB - This paper describes the preparation and optimization of the analytical properties of fluorescence-based submicrometric chloride ion sensing lipobeads. Fluorescence sensing lipobeads are polystyrene nanoparticles that are coated with a phospholipid membrane that contains a fluorescent indicator for a targeted analyte. In this study, the halide-specific fluorescence dye, lucigenin, was immobilized into the phospholipid membrane of the lipobeads to enable chloride ion detection. The fluorescence intensity of lucigenin decreases with increasing chloride ion concentration due to dynamic quenching. Lipobeads that contained only lucigenin were ineffective as chloride ion sensors due to poor partition of the water-soluble lucigenin molecules into the phospholipid membrane and high leakage rate of immobilized lucigenin molecules to the aqueous solution. To stabilize the chloride ion sensing lipobeads we coimmobilized hexadecanesulfonate molecules into the phospholipid membrane. The formation of ion pairs between hexadecanesulfonate and lucigenin decreased the hydrophilicity of the dye, increased its partition rate into the membrane, increased the brightness of the particles, and significantly decreased the leakage rate of the hydrophobic ion pair from the membrane to the solution. To further improve their chloride ion sensitivity, we also immobilized the chloride ionophore [9] mercuracarborand-3 into the lipobead membrane. The study resulted in a unique submicrometric chloride ion sensor, which is suitable for chloride ion measurements in biological fluids. PMID- 14750849 TI - Spatially resolved analysis of small particles by confocal Raman microscopy: depth profiling and optical trapping. AB - Raman microscopy is a powerful method to provide spatially resolved information about the chemical composition of materials. With confocal collection optics, the method is well suited to the analysis of small particles, either resting on a surface or optically trapped at a laser focus, where the confocal collection volume optimizes the signal from the particle. In this work, the sensitivity and spatial selectivity of detecting Raman scattering from single particles was determined as a function of particle size. An inverted confocal Raman microscope was used to acquire spectra of individual surface-bound and optically trapped polystyrene particles with sizes ranging between 200 nm and 10 microm. The particles are in contact with aqueous solution containing perchlorate ion that served as a solution-phase Raman-active probe to detect interferences from the surrounding medium. The collection volume is scanned through single particles that are attached to the surface of the coverslip, and the sensitivity and selectivity of detection are measured versus particle size. The results compare favorably with a theoretical analysis of the excitation profile and confocal collection efficiency integrated over the volumes of the spherical particles and the surrounding solution. This analysis was also applied to the detection of particles that are optically trapped and levitated above the surface of the coverslip. The results are consistent with the optical trapping of particles at or near the excitation beam focus, which optimizes excitation and selective collection of Raman scattering from the particle. PMID- 14750850 TI - Characterization of microorganisms using UV resonance Raman spectroscopy and chemometrics. AB - The past decade has seen an increased interest in the application of several physicochemical analytical techniques for the rapid detection and identification of microorganisms. We report the development of UV resonance Raman (UVRR) spectroscopy for the reproducible acquisition of information rich Raman fingerprints from endospore-forming bacteria belonging to the genera Bacillus and Brevibacillus. UVRR was conducted at 244 nm, and spectra were collected in typically 60 s. Cluster analyses of these spectra showed that UVRR spectroscopy could be used to discriminate between these microorganisms to species level, and the clustering pattern from this phenotypic classification was highly congruent with phylogenetic trees constructed from 16S rDNA sequence analysis. Therefore, we conclude that UVRR spectroscopy when coupled with chemometrics constitutes a powerful approach to the characterization and speciation of microorganisms. PMID- 14750851 TI - Comparison of surface-enhanced resonance Raman scattering from unaggregated and aggregated nanoparticles. AB - The effect of excitation frequency and state of aggregation on the sensitivity obtained in ultratrace analysis using colloidal suspensions of silver nanoparticles and surface-enhanced resonance Raman scattering (SERRS) detection is explored to define suitable conditions for quantitative analysis. Two structurally similar dyes, only one of which causes aggregation, were used as analytes without the use of external aggregating agents, thus simplifying the surface chemistry and removing a major source of error. Addition of the nonaggregating dye caused no change in particle charge or size and no time dependent aggregation as measured by zeta potential and particle size analysis. The most intense single-particle scattering was obtained using excitation at the wavelength of the plasmon resonance. Molecular resonance added approximately 2 orders of magnitude in sensitivity. Addition of the aggregating dye caused a reduction in surface charge of the particles and initiated a time-dependent aggregation process. However, constant SERRS with time is obtained at some excitation wavelengths probably because a constant number of clusters active at these wavelengths is maintained in the dynamic aggregation process. The additional enhancement caused by aggregation and molecular resonance is spread over a range of excitation frequencies. However, electronic spectra suggested that plasmon resonance enhancement would be effective at the longest wavelength of excitation used (785 nm), but there was a significant drop in intensity this far away from the absorbance maximum of the dye (429 nm). Thus, sensitive analysis using suspensions of single nanoparticles is feasible provided the excitation frequency used is close to that of the plasmon resonance frequency. Aggregation adds only an enhancement of approximately 6 in the experiments performed since only some particles in aggregates will have an active plasmon at any one wavelength, but the range of excitation wavelengths at which good enhancement is obtained is wider giving more flexibility if more complexity. PMID- 14750852 TI - Reagentless identification of single bacterial spores in aqueous solution by confocal laser tweezers Raman spectroscopy. AB - We demonstrate that optical trapping combined with confocal Raman spectroscopy using a single laser source is a powerful tool for the rapid identification of micrometer-sized particles in an aqueous environment. Optical trapping immobilizes the particle while maintaining it in the center of the laser beam path and within the laser focus, thus maximizing the collection of its Raman signals. The single particle is completely isolated from other particles and substrate surfaces, therefore eliminating any unwanted background signals and ensuring that information is collected only from the selected, individual particle. In this work, an inverted confocal Raman microscope is combined with optical trapping to probe and analyze bacterial spores in solution. Rapid, reagentless detection and identification of bacterial spores with no false positives from a complex mixed sample containing polystyrene and silica beads in aqueous suspension is demonstrated. In addition, the technique is used to analyze the relative concentration of each type of particle in the mixture. Our results show the feasibility for incorporating this technique in combination with a flow cytometric-type scheme in which the intrinsic Raman signatures of the particles are used instead of or in addition to fluorescent labels to identify cells, bacteria, and particles in a wide range of applications. PMID- 14750853 TI - Infrared-visible sum frequency generation investigation of Cu corrosion inhibition with benzotriazole. AB - Infrared-visible sum frequency generation spectroscopy is used to investigate the corrosion inhibitor benzotriazole (BTAH) adsorbed on Cu(100) and Cu(111) in acidic solution. Potential-dependent in situ spectra indicate that the adsorbed molecule is the benzotriazole anion (BTA-) at all potentials investigated. The Cu(100) surface is shown to form an ordered adlayer at all potentials probed, while the Cu(111) face is shown to be disordered at negative potentials, but to order with applied positive potential. The ordered adlayer is shown to consist of the BTA- in two configurations, one coordinated to the surface and Cu+ ions in solution and the other coordinated only to the surface. The BTA- coordinated to Cu+ is shown to be more stable with respect to Cl- addition than BTA- coordinated to only the surface. This study demonstrates the viability of using sum frequency generation to study corrosion inhibition in situ. PMID- 14750854 TI - Noninvasive continuous monitoring of physiological glucose using a monosaccharide sensing contact lens. AB - We have tested the feasibility of tear glucose sensing using a daily, disposable contact lens embedded with boronic acid-containing fluorophores as a potential alternative to current invasive glucose-monitoring techniques. Our findings show that our approach may, indeed, be suitable for the continuous monitoring of tear glucose levels in the range 50-500 microM, which track blood glucose levels that are approximately 5-10-fold higher. We compare the response of the boronic acid probes in the contact lens to solution-based measurements and can conclude that both the pH and polarity within the contact lens need to be considered with respect to choosing/designing and optimizing glucose-sensing probes for contact lenses. PMID- 14750855 TI - Selective detection and identification of sugar nucleotides by CE-electrospray-MS and its application to bacterial metabolomics. AB - A novel method employing CE-ESMS and precursor ion scanning was developed for the selective detection of nucleotide-activated sugars. By using precursor ion scanning for fragment ions specific to the different nucleotide carriers, i.e., ions at m/z 322 for cytidine monophosphate, m/z 323, 385, and 403 for uridine diphosphate, m/z 362, 424, and 442, for guanosine diphosphate, and m/z 346, 408, and 426 for adenosine diphosphate, it was possible to selectively detect sugar nucleotides involved in the biosynthesis of glycoconjugates such as glycoproteins and lipopolysaccharides. Enhancement of sensitivity was achieved using N-(2 hydroxyethyl)piperazine-N'-(2-ethanesulfonic acid) as a sample stacking buffer and provided detection limits between 0.2 and 3.8 pmol.mL(-)(1). The present CE ESMS method provided linear dynamic ranges over the concentrations 0.2-164 nM (r(2) = 0.952-0.997) for different nucleotide sugar standards. The application of this method is demonstrated for the identification of intracellular pools of sugar nucleotides in wild type and isogenic mutants from the bacterial pathogen Campylobacter jejuni. By using product ion scanning (with and without front-end collision-induced dissociation), it was possible to determine the precise nature of unexpected sugar nucleotides involved in the biosynthesis of pseudaminic acid, a sialic acid-like sugar previously observed on the flagellin of some pathogenic bacteria. PMID- 14750856 TI - Absolute configuration of monodentate phosphine ligand enantiomers on Cu(111). AB - Scanning tunneling microscopy (STM) has been employed to investigate the chirality of monophosphine compounds that are highly efficient chiral ligands in transition-metal-catalyzed organic transformations. The absolute configuration of 1-(2-diphenyphosphino-1-naphthyl)isoquinoline enantiomers with axial chirality was discriminated directly by the "marker" group, PPh(2) substitutes. Although the two enantiomer molecules adsorb on a Cu(111) surface and form well-defined (4 x 4) structures, the positions of PPh(2) substitutes in the molecular adlayers are different. The mirror symmetry between two adlayers is demonstrated. On the basis of STM results, structural models are proposed to interpret the chiral adsorption. The results presented here supply a straightforward method for axial chirality analysis in adsorbed adlayers by STM. PMID- 14750857 TI - Improvement of homogeneity of analytical biodevices by gene manipulation. AB - Homogeneity is proposed for evaluation of the quality of analytical biodevices, such as biosensors and biochips. As a demonstration, glucose oxidase (GOx) was modified at its C-terminal with a linker peptide with a cysteine residue at the end. The fusion structure (GOx-linker-cysteine) enables the enzyme to immobilize on gold surfaces with a Cys-S-Au bond or to immobilize on a silanized glass surface via disulfide chemistry. With this fusion structure, the enzyme can be anchored onto the substrate with well-controlled orientation, thus forming a homogeneous biological layer on biodevices. The linker peptide between GOx and the cysteine acts as a spacer to reduce the steric hindrance caused by the bulky body of the enzyme. Biochemistry experiments showed that this genetically modified glucose oxidase (shortened to GOxm) retained most of its catalytic characteristics, with K(m) and K(cat) similar to those of the wild-type GOx. Electrochemistry experiments showed that GOxm-modified electrode gave higher and more stable current responses than the electrode modified with GOx which has no free -SH on its surface. The coefficients of variation (used for evaluation of the interchangeability of the enzyme device from the same batch preparation) were 9.5% for the GOxm gold electrode and 20.0% for the GOx gold electrode and the GOxm oxygen electrode. The relative errors (used for evaluation of the precision of the individual enzyme device) were 2.9% for the GOxm gold electrode, 12.0% for the GOx gold electrode, and 11.2% for the GOxm oxygen electrode. Atomic force microscopy images revealed that GOxm formed a self-assembled monolayer in a hexagonal-like lattice packing arrangement on the gold surface, while GOx formed multilayer assembling or aggregated particles. The homogeneity of the protein chips, the GOxm array that was prepared through -S-S- formation, and the GOx array that was prepared through nonspecific adsorption was evaluated. The coefficients of variation, calculated with the signal level of all dots, were 5.4% for the GOxm array and 81.8% for the GOx array. All experimental results pointed to the fact that the homogeneity of the analytical biodevices could be considerably improved by using the proposed method. PMID- 14750858 TI - Analysis of the performance of a flow reactor for use with microcolumn HPLC. AB - Postcolumn derivatization reactions can be used to improve detector sensitivity or selectivity. The advantages of capillary chromatography for trace analysis could be augmented if there were postcolumn reactors suitable for microchromatographic systems. However, postcolumn derivatization is a challenge because of the small peak volumes associated with capillary columns. We have developed a postcolumn flow reactor from microchannels formed in fluorinated ethylene propylene and 50-microm fused-silica tubing for use with capillary HPLC analyses. Theoretical and experimental evidence show that the reactor, which operates in the Taylor dispersion regime, enables contact of analyte and derivatization streams purely by diffusion. Reactor lengths as short as 2 cm allow formation of copper(II)-peptide complexes that are detected electrochemically at a carbon fiber microelectrode. The reactor has been used with 100-microm-i.d. columns with insignificant effects (i.e., <3%) on peak band spreading. Theoretical calculations indicate that even smaller i.d. columns can be used with little effect on chromatographic resolution. PMID- 14750859 TI - Automated microarray system for the simultaneous detection of antibiotics in milk. AB - A parallel affinity sensor array (PASA) for the rapid automated analysis of 10 antibiotics in milk is presented, using multianalyte immunoassays with an indirect competitive ELISA format. Microscope glass slides modified with (3 glycidyloxypropyl)trimethoxysilane were used for the preparation of hapten microarrays. Protein conjugates of the haptens were immobilized as spots on disposable chips, which were processed in a flow cell. Monoclonal antibodies against penicillin G, cloxacillin, cephapirin, sulfadiazine, sulfamethazine, streptomycin, gentamicin, neomycin, erythromycin, and tylosin allowed the simultaneous detection of the respective analytes. Antibody binding was detected by a second antibody labeled with horseradish peroxidase generating enhanced chemiluminescence, which was recorded with a sensitive CCD camera. All liquid handling and sample processing was fully automated, and one analysis was carried out in milk within less than 5 min. The detection limits ranged from 0.12 (cephapirin) to 32 microg/L (neomycin). Penicillin G could be detected at the maximum residue limit (MRL); the detection limits for all other analytes were far below the respective MRLs. The PASA system proved to be the first immunochemical biosensor platform having the potential to test for numerous antibiotics in parallel, such being of considerable interest for the control of milk in the dairy industry. PMID- 14750860 TI - Capillary electrophoretic separation of nuclei released from single cells. AB - We report here the first capillary electrophoresis analysis of intact nuclei released on-column from single cells. Expression of the nuclear-targeted protein nuDsRed2 and the plasma membrane-bound farnesylated enhanced green fluorescent protein in cultured human DeltaH2-1 cells allowed fluorescent monitoring of the fate of these subcellular compartments upon injection of a single cell into the separation capillary. On-column treatment with digitonin allowed for the separation of the plasma membrane from the nucleus as indicated by their selective laser-induced fluorescence detection in two separate spectral regions. The data suggest that less than 0.1% of the plasma membrane remains bound to individually detected nuclei. In digitonin-treated cells, the electropherograms consisted of a prominent fluorescent peak attributed to nuDsRed2 localized to the nucleus and a collection of weakly fluorescent events (barely distinguishable from scattering) that seem to indicate additional localization of this protein to other subcellular regions. Taken together, this report points to the feasibility of studying intact organelles released from a single mammalian cell by capillary electrophoresis, which is a prerequisite to understanding the relevance of subcellular heterogeneity in biological systems. PMID- 14750861 TI - Chiral morphing and enantiomeric quantification in mixtures by mass spectrometry. AB - A novel mass spectrometric method is introduced for rapid and accurate chiral quantification by examining a tetracoordinated transition metal complex into which a reference and a fixed ligand are incorporated simultaneously with the analyte. Chiral analysis is performed by measuring the dissociation kinetics of these trimeric cluster ions [(M(II) + L(fixed) - H)(ref)(An)]+ (M(II) = a transition metal ion, L(fixed) = chiral peptide fixed ligand, ref = chiral reference ligand, and An = chiral analyte) in an ion trap mass spectrometer. The ratio of the product ion branching ratios measured when a pair of pure chiral fixed ligands and chiral reference ligands (/ref(D) and /ref(L); or /ref(L) and /ref(D)) are employed in separate experiments is related, via the kinetic method formalism, to the enantiomeric composition of the chiral mixture. This fixed ligand quotient ratio (QR(fixed)) is logarithmically proportional to enantiomeric purity allowing construction of a calibration curve for chiral analysis when the analyte is only available in one form of known optical purity. There are reciprocal relationships when switching the chirality of the fixed/reference ligands. Improved quantification accuracy (due to simplified dissociation kinetics) and ready construction of two or more single-point calibration curves allow data to be cross-checked and represent an advantage of this approach. These features and the matrix tolerance of the kinetic method are demonstrated using the QR(fixed) method for determinations of enantiomeric excess of the drug DOPA in the presence of the co-drug compound L-carbidopa. The chiral selectivity of DOPA was found to vary from 0.0581 to 0.337 using this method, depending on the choices of fixed-ligand and reference chirality. The average relative errors are less than 1.2%. The potential of chiral morphing (changing chiral centers in the ligands) to further refine the chiral interactions and hence to maximize chiral recognition is shown. PMID- 14750862 TI - Analysis of protein solvent accessible surfaces by photochemical oxidation and mass spectrometry. AB - Protein surfaces are important in most biological processes, including protein protein interactions, enzymatic catalysis, and protein-ligand binding. We report a method in which hydroxyl radicals generated by a rapid-UV irradiation of a 15% hydrogen peroxide solution were utilized to oxidize specific amino acid side chains of two model proteins (lysozyme, beta-lactoglobulin A), according to the residues' chemical reactivities and the solvent accessibility of the reactive carbons and sulfurs in the residue. Oxidized peptides generated by tryptic digestion were identified by electrospray-Fourier transform mass spectrometry. The specific sites of the stable modification were then identified by reverse phase liquid chromatography coupled to quadropole ion trap tandem mass spectrometry. The solvent accessibility of the residue was shown to directly affect the rate of oxidation by this method (with the exception of methionine), supporting its use as a rapid measure of the solvent accessibility of specific residues, and in some cases, individual atoms. PMID- 14750863 TI - Multiplexed toxin analysis using four colors of quantum dot fluororeagents. AB - Quantum dots (QDs) have the potential to simplify the performance of multiplexed analysis. In this work, we prepared bioinorganic conjugates made with highly luminescent semiconductor nanocrystals (CdSe-ZnS core-shell QDs) and antibodies to perform multiplexed fluoroimmunoassays. Sandwich immunoassays for the detection of cholera toxin, ricin, shiga-like toxin 1, and staphylococcal enterotoxin B were performed simultaneously in single wells of a microtiter plate. Initially the assay performance for the detection of each toxin was examined. We then demonstrated the simultaneous detection of the four toxins from a single sample probed with a mixture of all four QD-antibody reagents. Using a simple linear equation-based algorithm, it was possible to deconvolute the signal from mixed toxin samples, which allowed quantitation of all four toxins simultaneously. PMID- 14750864 TI - Electrical detection of viral DNA using ultramicroelectrode arrays. AB - A fully electrical array for voltammetric detection of redox molecules produced by enzyme-labeled affinity binding complexes is shown. The electronic detection is based on ultramicroelectrode arrays manufactured in silicon technology. The 200-microm circular array positions have 800-nm-wide interdigitated gold ultramicroelectrodes embedded in silicon dioxide. Immobilization of oligonucleotide capture probes onto the gold electrodes surfaces is accomplished via thiol-gold self-assembling. Spatial separation of probes at different array positions is controlled by polymeric rings around each array position. The affinity bound complexes are labeled with alkaline phosphatase, which converts the electrochemically inactive substrate 4-aminophenyl phosphate into the active 4-hydroxyaniline (HA). The nanoscaled electrodes are used to perform a sensitive detection of enzyme activity by signal enhancing redox recycling of HA resulting in local and position-specific current signals. Multiplexing and serial readout is realized using a CMOS ASIC module and a computer-controlled multichannel potentiostat. The principle of the silicon-based electrical biochip array is shown for different experimental setups and for the detection of virus DNA in real unpurified multiplex PCR samples. The fast and quantitative electronic multicomponent analysis for all kinds of affinity assays is robust and particle tolerant. PMID- 14750865 TI - Direct analysis of an oligomeric hindered amine light stabilizer in polypropylene materials by MALDI-MS using a solid sampling technique to study its photostabilizing action. AB - A novel method for the direct analysis of small amounts of an oligomeric hindered amine light stabilizer (HALS) occluded in polypropylene (PP) material was developed to study its photostabilizing action on the basis of matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) using a solid sampling technique while avoiding troublesome solvent extraction. In this sampling protocol, the powdered mixture of PP composite sample containing trace amounts of an oligomeric HALS, Adekastab LA-68LD (MW = 1900), and the matrix reagent (dithranol) was spotted on the sample plate, then ion exchanged water was deposited onto the mixture to make a suspension, and finally, the dried mixture adhered on the plate was subjected to MALDI-MS measurement. On the mass spectrum thus obtained by the solid sampling MALDI, the molecular ions of the HALS desorbed from the PP composite were clearly observed as three major series of the HALS components in the range up to about m/z 7000 with little interference by the PP substrate and the other additives. Moreover, in the MALDI-MS spectra for the UV-exposed sample, the satellite peaks around the major HALS components proved to enhance significantly, reflecting the oxidized HALS species at the tetramethylpiperidine units to cause the photostabilizing action. In addition, hydrolyzed HALS species were also observed for the irradiated sample. These results suggest that not only the oxidation reaction but also the hydrolysis or decomposition of the oligomeric HALS components competitively proceed in the PP composites during UV exposure. PMID- 14750866 TI - Analysis of multiple endocrine disruptors in environmental waters via wide spectrum solid-phase extraction and dual-polarity ionization LC-ion trap-MS/MS. AB - An analytical method for the determination of 35 endocrine disrupting chemicals (EDCs) present in the aquatic environment was developed and validated. The procedure includes an off-line solid-phase extraction of 500-mL water samples using wide-spectrum polymer packing material combined with two LC-ESI-MS/MS runs, in negative and positive ionization modes. Limits of quantitation were established between 0.1 and 20.0 ng/L. Satisfactory recoveries were obtained ranging from 80.1 to 110.2%. Calibration, using deuterated internal standardization, was performed by linear regression analysis. Linearity (R(2) > 0.99) was demonstrated over individually specified ranges using seven calibration points for each analyte. Intrabatch and interbatch precision, as well as accuracy (n = 5), were investigated at low, medium, and high concentrations. Precision for all compounds, expressed as the RSD, proved to be less than 17.8 and 20.0%, respectively, for intra- and interbatch. Accuracy, expressed as the mean recovery, was between 83.1 and 108.4% at all concentrations. Stability experiments showed no significant loss or deterioration for any of the analytes. Finally, the method was applied on real samples. PMID- 14750867 TI - Development and characterization of an aerosol time-of-flight mass spectrometer with increased detection efficiency. AB - This paper describes the development and characterization studies of a more efficient aerosol time-of-flight mass spectrometer (ATOFMS), showing results for the on-line detection and determination of the size and chemical composition of single fine (100-300 nm) and ultrafine (<100 nm) particles. An aerodynamic lens inlet was implemented, replacing the converging nozzle inlet used on conventional ATOFMS instruments. In addition, the light scattering region was modified to enhance the scattering signals for smaller particles. Polystyrene latex spheres (PSL) with aerodynamic diameters ranging from 95 to 290 nm were used to characterize the particle sizing efficiency (product of particle transmission efficiency and particle scattering efficiency), particle detection efficiency (product of particle sizing efficiency and particle hit rate), and particle beam profile and perform instrument calibration. At number concentrations of <20 particles/cm(3), the particle sizing efficiencies were determined to be approximately 0.5% for 95 nm and approximately 47% for 290-nm PSL particles, while the particle detection efficiencies were measured to be approximately 0.3% for 95 nm and 44% for 290-nm PSL particles. This represents a significant increase (i.e., at least 3 orders of magnitude) in detection efficiencies for smaller particles over the conventional ATOFMS. In addition, the beam profiles for PSL particles of various sizes were measured in the ion source of the mass spectrometer and follow a Gaussian distribution with a full width at half-maximum of approximately 0.35 mm. The resulting higher detection efficiencies allow ATOFMS to obtain higher temporal resolution measurements of the composition of fine and ultrafine individual particles as demonstrated in initial ambient measurements in La Jolla, CA. At typical ambient particle number concentrations of 10(2)-10(3) particles/cm(3), approximately 30 000 particles with aerodynamic diameters of <300 nm were detected with average 24-h hit rates of 30% for particles between 50 and 300 nm. This advancement, allowing for high temporal resolution measurements of the composition of smaller particles with higher efficiency, adds to a growing number of instruments that can chemically characterize individual fine and ultrafine particles, with the goal of providing new insights into a number of areas including environmental and material sciences, health effects studies, industrial hygiene, and national security. PMID- 14750868 TI - Whole protein dissociation in a quadrupole ion trap: identification of an a priori unknown modified protein. AB - A protein mixture derived from a whole cell lysate fraction of Saccharomyces cerevisiae, which contains roughly 19 proteins, has been analyzed to identify an a priori unknown modified protein using a quadrupole ion trap tandem mass spectrometer. Collection of the experimental data was facilitated by collision induced dissociation and ion/ion proton-transfer reactions in multistage mass spectrometry procedures. Ion/ion reactions were used to manipulate charge states of both parent ions and product ions for the purpose of concentrating charge into the parent ion of interest and to reduce the product ion charge states for determination of product ion mass and abundance. The identification of the protein was achieved by matching the uninterpreted product ion spectrum against protein sequence databases with varying degrees of annotation, coupled with a scoring scheme weighted for the relative abundances of the experimentally observed product ions and the frequency of fragmentations occurring at preferential sites. The protein was identified to be an acetylated yeast heat shock protein, HS12_Yeast (11.6 kDa), with the initiating methionine residue removed. This constitutes the first example of the identification of an a priori unknown protein that is not present in an annotated protein database using a "top down" approach with a quadrupole ion trap. This example illustrates the utility of relatively low cost instrumentation with modest mass analysis characteristics for the identification of modified proteins without recourse to enzymatic digestion. It also illustrates how experimental data can be used interactively with protein databases when the modified protein of interest is not initially present in the database. PMID- 14750869 TI - Fluorogenic derivatization reagents suitable for isolation and identification of cysteine-containing proteins utilizing high-performance liquid chromatography tandem mass spectrometry. AB - The fluorogenic derivatization reagents with a positive charge, 4 (dimethylaminoethylaminosulfonyl)-7-chloro-2,1,3-benzoxadiazole (DAABD-Cl) and 7 chloro-2,1,3-benzoxadiazole-4-sulfonylaminoethyltrimethylammonium chloride (TAABD Cl), are proposed for use in proteomics studies. Following derivatization of protein mixtures with these reagents, a series of standard processes of isolation, digestion, and identification of the proteins were performed utilizing high-performance liquid chromatography-fluorescence detection and tandem mass spectrometry with the probability-based protein identification algorithm. Both DAABD and TAABD derivatives were detected fluorometrically at the femtomole level and showed more than 100-fold improvement in sensitivity compared to the underivatized original compounds with an electrospray ionization ion trap mass spectrometer analysis. The modification of the MASCOT database search system memorized with the fragment information of a DAABD-attached Cys residue allowed the identification of the proteolytic peptide fragments of the derivatized bovine serum albumin (BSA) with an estimated 38% sequence coverage of BSA. Utilizing DAABD-Cl as a derivatization reagent, identification of several proteins was also possible in a soluble extract of Caenorhabditis elegans (10 microg of protein). Consequently, for identification of proteins in the complex matrixes of proteins, DAABD-Cl could be a more appropriate reagent than ammonium 7-fluoro-2,1,3 benzoxadiazole-4-sulfonate as reported previously. PMID- 14750870 TI - Derivatization of chiral amino acids in supercritical carbon dioxide. AB - A new method is proposed to perform the derivatization of chiral amino acids occurring in complex samples using supercritical carbon dioxide as both the reaction medium and the agent used to extract the obtained derivatives prior to accomplishing the subsequent enantiomeric chromatographic analysis. The derivatization step under supercritical conditions involves the esterification of the carboxyl group and the acylation of the amino group of the amino acids without using a catalyst. A Chirasil-L-Val capillary column enabled the separation of the D- and L-forms of the amino acids as their N(O) pentafluoropropionyl 1-propyl esters. Relative standard deviation values obtained from the gas chromatographic analysis for the derivatized amino acids ranged from 5 to 15%. PMID- 14750871 TI - Integration of isoelectric focusing with parallel sodium dodecyl sulfate gel electrophoresis for multidimensional protein separations in a plastic microfluidic [correction of microfludic] network. AB - An integrated protein concentration/separation system, combining non-native isoelectric focusing (IEF) with sodium dodecyl sulfate (SDS) gel electrophoresis on a polymer microfluidic chip, is reported. The system provides significant analyte concentration and extremely high resolving power for separated protein mixtures. The ability to introduce and isolate multiple separation media in a plastic microfluidic network is one of two key requirements for achieving multidimensional protein separations. The second requirement lies in the quantitative transfer of focused proteins from the first to second separation dimensions without significant loss in the resolution acquired from the first dimension. Rather than sequentially sampling protein analytes eluted from IEF, focused proteins are electrokinetically transferred into an array of orthogonal microchannels and further resolved by SDS gel electrophoresis in a parallel and high-throughput format. Resolved protein analytes are monitored using noncovalent, environment-sensitive, fluorescent probes such as Sypro Red. In comparison with covalently labeling proteins, the use of Sypro staining during electrophoretic separations not only presents a generic detection approach for the analysis of complex protein mixtures such as cell lysates but also avoids additional introduction of protein microheterogeneity as the result of labeling reaction. A comprehensive 2-D protein separation is completed in less than 10 min with an overall peak capacity of approximately 1700 using a chip with planar dimensions of as small as 2 cm x 3 cm. Significant enhancement in the peak capacity can be realized by simply raising the density of microchannels in the array, thereby increasing the number of IEF fractions further analyzed in the size-based separation dimension. PMID- 14750872 TI - pH gradient reversed-phase HPLC. AB - pH gradient HPLC is reported, which is a new original mode of reversed-phase high performance liquid chromatography applicable to ionogenic analytes. The method consists of programmed increase during the chromatographic run of the eluting strength of the mobile phase with respect to the acid/base analytes separated. Unlike the well-established conventional gradient HPLC, where the eluting power of the mobile phase is increased with time due to the increasing content of organic modifier, in the pH gradient HPLC that is realized by linearly increasing (in the case of acids) or decreasing (in the case of bases) the pH of the eluent of a fixed organic modifier content, thus providing functional increase in the degree of analyte dissociation and, hence, a decrease in its retention. The pH gradient mode has typical features of gradient HPLC, such as reduced peak width and minimized peak-tailing due to peak compression, which is especially advantageous in the case of organic base analytes. It may be of special value for separation of those analytes which are susceptible to the higher concentrations of organic solvents, as many bioanalytes are. A theory of the pH gradient HPLC has been elaborated, and its full mathematical formalistic is presented step by step in a comprehensive manner. Although fundamental relationships at the basis of pH gradient HPLC are more complex than in the case of the organic gradient variant, the resulting mathematical model is easily manageable. Its applicability to predict changes in retention and separation of test mixtures of analytes accompanying the changes in chromatographic conditions has been demonstrated experimentally in both gradient and isocratic HPLC. The proposed model supplies a rational basis for modifications of eluent pH aimed at optimization of separations and for convenient assessment of chromatographically relevant physicochemical parameters of analytes, such as pK(a). PMID- 14750873 TI - Probing the interaction of benzo[a]pyrene adducts and metabolites with monoclonal antibodies using fluorescence line-narrowing spectroscopy. AB - A new approach for studying antibody-antigen interactions of DNA adducts and metabolites of polycyclic aromatic hydrocarbons (PAHs) is demonstrated in which fluorescence line-narrowing spectroscopy (FLNS) is used. It is based on the fact that in an FLN spectrum the relative intensities of the line-narrowed bands (that correspond to the excited-state vibrations) are, in general, strongly dependent on the local environment of the fluorophore. Information on the nature of the interactions can be obtained by comparing the FLN spectra of the antigen-antibody complexes to the spectra of the antigen in different types of solvents (H bonding, aprotic, and pi-electron-containing solvent molecules) recorded under the same conditions. The antigens used were the DNA adduct 7-(benzo[a]pyren-6 yl)guanine (BP-6-N7Gua) and the metabolite (+)-trans-anti-7,8,9,10 benzo[a]pyrenetetrol (BP-tetrol) of benzo[a]pyrene; two monoclonal antibodies (MAbs) have been developed to selectively bind these compounds. It is shown that, for BP-tetrol, H-bonding solvents have a pronounced effect on the FLN spectra. The presence of pi electrons in the solvent molecules results in relatively small but still significant changes in the spectra. When BP-tetrol is bound to its MAb, however, neither of these effects is observed; its spectrum is very similar to the one obtained with an aprotic solvent, methylcyclohexane. Therefore, we can conclude that this MAb has an internal binding site in which the interaction with BP-tetrol is of a hydrophobic character. For BP-6-N7Gua, however, there is a strong effect of the presence of pi electrons in the solvent molecules. The FLN spectrum of this antigen bound to its MAb is very similar to its spectrum in acetone, indicating that pi-pi interactions play an important role in the binding. PMID- 14750874 TI - Use of excess solid-phase capacity in immunoassays: advantages for semicontinuous, near-real-time measurements and for analysis of matrix effects. AB - A flow-based immunoassay system using solid-phase particles with high binding capacity was used for semicontinuous, near-real-time, measurement of 17beta estradiol (E2). The high binding capacity of the solid phase was exploited to enable (i) a quantitative determination of E2 concentration, based on rate of accumulation of fluorescently labeled anti-E2 antibody on the solid phase, and (ii) the use of a single solid phase for more than a dozen competitive binding measurements. The high binding capacity of the solid phase also permitted the immobilization of a second capture antigen. Biotin was immobilized as a second antigen and used to evaluate a biotin anti-biotin system as a control for matrix effects in the E2 immunoassay. In phosphate-buffered saline, E2 could be quantified (in the range of 10-1000 pM) by using either the summation or ratio of the signals from the labeled anti-E2 and anti-biotin antibody in the presence of biotin at a constant concentration. The same referencing system was applied to estimate the matrix effects in selected environmental samples. Matrix effects that inhibited the binding of the anti-E2 antibody to the solid phase led to false positive responses, but these matrix effects could be identified and partially corrected using the response from the anti-biotin antibody. PMID- 14750875 TI - Automated enzymatic assays in a renewable fashion using the multisyringe flow injection scheme with soluble enzymes. AB - In this paper, a novel flowing stream scheme based upon the multisyringe flow injection (MSFI) technique is presented as a powerful tool to perform automated enzymatic assays. The exploitation of enzymes in homogeneous phase circumvents typical drawbacks associated with the commonly used packed-bead or open tubular permanent columns, namely, malfunctions of the reactor, carryover effects, flow resistance, loss of binding sites, large reagent consumption, and use of harmful organic solvents during immobilization procedures. The proposed MSFI system is able to handle minute volumes of soluble enzymes and accommodate reactions with divergent kinetic and pH demands, as demonstrated via the indirect chemiluminescence determination of trace levels of glucose. The procedure is based on the on-line glucose oxidase-catalyzed oxidation of beta-glucose in homogeneous phase to beta-glucono-delta-lactone and hydrogen peroxide. Subsequently, the generated oxidant merges downstream with an alkaline slug of 3 aminopthalhydrazide and a metal-catalyst zone (viz., Co(II)) at a total flow rate as high as 72 mL/min aiming to warrant maximum light collection from the fast CL reaction. Under optimum conditions for both sequentially occurring reactions, a glucose concentration as low as 90 microg/L may be easily detected at a 1000-fold photomultiplier gain. A second-order polynomial regression equation of light emission versus substrate concentration is found over the range 90 microg/L-2.7 mg/L glucose, although a maximum concentration of 180 mg/L may be determined by suitable gain selection without requiring manifold reconfiguration. An injection throughput of 20 h(-1), a repeatability better than 2.5% at the 1 mg/L level, and a 3sigma detection limit of 72 microg/L are the analytical features of the designed analyzer. The proposed approach was applied to the analysis of ultralow glucose content soft drinks as well as fruit juices suitable for diabetic consumers. The accuracy was assessed using the spectrophotometric batch glucose Trinder method as an external reference methodology for the determination of the target species in parenteral solutions. PMID- 14750876 TI - Attenuated total reflectance FTIR detection and quantification of low concentrations of aqueous polyatomic anions. AB - Development of a new quantitative method for determining low concentrations of aqueous polyatomic anions using attenuated total reflectance (ATR) FTIR spectroscopy is described. Evaporated thin-film coatings of anion-selective tetraalkylated ferrocenium salts were applied to the surface of ATR crystals, which enabled anion detection limits to be lowered up to 23 000-fold below those achieved using the commercially available spectrometer with identical uncoated ATR crystals. Linear calibration curves based on d(absorbance)/dt, which is related to the rate of anion exchange in the thin film, were established in the 0.04-30 microM range. Limits of detection (10-min analyses) for perchlorate, chlorate, trifluoromethanesulfonate, perfluoro-n-butanesulfonate, perfluoro-n octanesulfonate, tetrafluoroborate, hexafluorophosphate, and pinacolylmethylphosphonate in aqueous solution were 0.03, 0.2, 0.05, 0.07, 0.06, 0.06, 0.6, and 0.7 microM, respectively, using the thin-film coatings. This simple detection/quantification method afforded good reproducibility with relatively fast detection times. PMID- 14750877 TI - Quartz crystal microbalance and infrared reflection absorption spectroscopy characterization of bisphenol A absorption in the poly(acrylate) thin films. AB - The absorption process of bisphenol A (BPA) in a number of poly(acrylate) thin films, such as poly(2-methoxyethyl acrylate) (PMEA), poly(ethyl acrylate) (PEA), poly(n-butyl methacrylate) (PBMA), and poly(methyl methacrylate) (PMMA), has been investigated by quartz crystal microbalance (QCM) and infrared reflection absorption spectroscopy (IRRAS) measurements. Both QCM and IRRAS measurements show that the BPA molecules absorb in PMEA, PEA, and PBMA thin films but not in PMMA thin film. The differences in the BPA absorption behavior are mainly attributed to the difference in the glass transition temperature (T(g)) between these polymers. This absorption behavior also depends on the BPA concentration and polymer film thickness. Furthermore, IRRAS characterization demonstrates that the hydrogen bonding is formed between the hydroxyl group in BPA and the carbonyl group in the poly(acrylate) thin films. BPA molecule absorbed in these polymer thin films can be removed by ethanol rinse treatment. By optimizing experimental conditions for the QCM electrode modified by PMEA thin film, detection limitation of approximately 1 ppb for BPA can be realized by the in situ QCM measurement. This method is expected to be a sensitive in situ detection way for trace BPA in the environmental study. PMID- 14750878 TI - A model for the quartz crystal microbalance frequency response to wetting characteristics of corrugated surfaces. AB - We consider the effect of surface roughness, and its unique wetting behavior, on the response of a quartz crystal microbalance (QCM) resonator operating in contact with a fluid. The rough surface is modeled as sinusoidally corrugated particular to the case of a fixed relationship between amplitude and periodicity, as would arise from polishing with monodisperse spherical particles. The penetration of fluid into the troughs of the corrugations and the resulting meniscus are determined as a competition between surface tension and compression of the trapped gas. Liquid contained below the corrugation peaks, but above the gas/liquid meniscus, is trapped and behaves as an ideal mass layer, contributing a frequency shift that adds to that arising from liquid entrainment. This model allows QCM responses on rough surfaces to be described as a function of liquid properties and contact angle. This permits responses on hydrophobic surfaces to be understood in terms of incomplete surface wetting. PMID- 14750879 TI - Development of a portable immunoextraction-reversed-phase liquid chromatography system for field studies of herbicide residues. AB - A portable system based on immunoextraction and reversed-phase HPLC was developed for the field analysis of herbicides in groundwater and surface water. Atrazine, simazine, and cyanazine were used as model analytes for this work. These were measured in water by using three coupled columns: an anti-atrazine antibody column for the selective extraction of these analytes, a reversed-phase precolumn for their reconcentration, and a reversed-phase analytical column for their separation. Various factors were considered in the optimization of this system, including the binding properties of the immunoextraction column, the effect of flow rate on the performance of each column, the selection of sample volume, and the choice of mobile phases for the RPLC columns. A typical analysis with this system allowed the injection of one sample every 7.5 min and provided results for all three of the tested herbicides in less than 10 min. In the analysis of atrazine alone, samples could be injected every 4 min and results were obtained within 8 min. There was good correlation between this technique and a comparable benchtop system. The lower limits of detection for the given analytes were approximately 0.2-0.25 microg/L, with a linear range that extended to 20 microg/L and a dynamic range that went up to at least 100 microg/L. The use of this technique in the field was demonstrated through applications that involved the development of time and location profiles for triazine herbicides in environmental samples. PMID- 14750880 TI - Mass distribution measurement of water-insoluble polymers by charge-reduced electrospray mobility analysis. AB - 1-Methyl-2-pyrrolidone (NMP) seeded with 5% trifluoroacetic acid is identified as a singular buffer, polar enough to produce fine electrospray drops, yet having excellent solubility for many industrial polymers such as polystyrene (PSR) and poly(methyl methacrylate) (PMMA). Four PSR mass standards (M = 9.2, 34.5, 68, and 170 kDa) with narrow mass distributions are electrosprayed from their solutions in this buffer. The high charge on the resulting ions is reduced to unity with a radioactive source, whereby their electrical mobility distributions, determined by a differential mobility analyzer, yield unambiguously their size distribution. Each standard produces (at high solution concentration) several mobility peaks associated with the formation of particles containing from one to six polymer molecules, used to establish a relation Z(M) between electrical mobility Z and polymer mass. Within the indeterminacy given by inaccuracies in the nominal masses of the standards, this relation indicates that the polymers form spherical balls with a density close to the bulk density of polystyrene, as seen previously with poly(ethylene glycol) chains. Good mobility spectra from the same buffer are also obtained for PMMA (M = 49 kDa). Because NMP is less conductive and contains more involatile impurities than common aqueous buffers, the electrospray ions formed tend to carry a small contaminant crust, which distorts the inferred mass distribution unless a high spray quality is achieved. PMID- 14750881 TI - Integration of knowledge-based metabolic predictions with liquid chromatography data-dependent tandem mass spectrometry for drug metabolism studies: application to studies on the biotransformation of indinavir. AB - Despite recent advances in the application of data-dependent liquid chromatography/tandem mass spectrometry (LC/MS/MS) to the identification of drug metabolites in complex biological matrixes, a prior knowledge of the likely routes of biotransformation of the therapeutic agent of interest greatly facilitates the detection and structural characterization of its metabolites. Thus, prediction of the [M + H]+ m/z values of expected metabolites allows for the construction of user-defined MS(n) protocols that frequently reveal the presence of minor drug metabolites, even in the presence of a vast excess of coeluting endogenous constituents. However, this approach suffers from inherent user bias, as a result of which additional "survey scans" (e.g., precursor ion and constant neutral loss scans) are required to ensure detection of as many drug related components in the sample as possible. In the present study, a novel approach to this problem has been evaluated, in which knowledge-based predictions of metabolic pathways are first derived from a commercial database, the output from which is used to formulate a list-dependent LC/MS(n) data acquisition protocol. Using indinavir as a model drug, a substructure similarity search on the MDL metabolism database with a similarity index of 60% yielded 188 "hits", pointing to the possible operation of two hydrolytic, two N-dealkylation, three N glucuronidation, one N-methylation, and several aromatic and aliphatic oxidation pathways. Integration of this information with data-dependent LC/MS(n) analysis using an ion trap mass spectrometer led to the identification of 18 metabolites of indinavir following incubation of the drug with human hepatic postmitochondrial preparations. This result was accomplished with only a single LC/MS(n) run, representing significant savings in instrument use and operator time, and afforded an accurate view of the complex in vitro metabolic profile of this drug. PMID- 14750882 TI - Normal-phase nanoscale liquid chromatography-mass spectrometry of underivatized oligosaccharides at low-femtomole sensitivity. AB - We here describe the online liquid chromatography (LC) electrospray ionization mass spectrometry (MS) of underivatized glycans using a nanoscale normal-phase amide column at a flow rate of 300 nL/min. Retention on the amide column is based on polar interactions of the oligosaccharide hydroxyl groups with the stationary phase, and thus, the retention time predictably increases with elongation of the oligosaccharide chain. The system is characterized by its high chromatographic resolution, which routinely allows the separation of isobaric structures. Separation of oligosaccharide mixtures over a 1-h range permits the detailed characterization of the different species by multiple ion selection and fragmentation steps using ion trap MS. The here presented miniaturization of the online-LC system to the nanoscale in combination with ion trap MS allows the detection of oligosaccharide species in a mixture at low-femtomole sensitivity. Online normal-phase nano-LC-MS of complex oligosaccharide mixtures further facilitates the sensitive and detailed structural analysis of oligosaccharides by overcoming the need for cumbersome and time-consuming derivatization procedures such as reductive amination for labeling with hydrophobic fluorophores or labeling with tritium. The method should be useful for the sensitive and quick analysis of glycosylation patterns and individual oligosaccharides from biotechnologically produced glycoproteins as well as scarcely available biological samples. PMID- 14750883 TI - Favorable effects of weak acids on negative-ion electrospray ionization mass spectrometry. AB - Despite widespread use in pharmacokinetic, drug metabolism, and pesticide residue studies, little is known about the factors governing response during reversed phase liquid chromatography coupled with negative-ion electrospray ionization (ESI(-)) mass spectrometry. We examined the effects of various mobile-phase modifiers on the ESI(-) response of four selective androgen receptor modulators using a postcolumn infusion system. Acetic, propionic, and butyric acid improved the ESI(-) responses of analytes to varying extents at low concentrations. Formic acid suppressed ionization, as did neutral salts (ammonium formate, ammonium acetate) and bases (ammonium hydroxide, triethylamine) under most conditions. Two modifiers (2,2,2-trifluoroethanol, formaldehyde) that produce anions with high gas-phase proton affinity increased ESI(-) responses. However, the concentrations of these modifiers required to enhance ESI(-) response were higher than that of acidic modifiers, which is a phenomenon likely related to their low pK(a) values. 2,2,2-Trifluoroethanol increased response of more hydrophobic compounds but decreased response of a more hydrophilic compound. Formaldehyde improved response of all the compounds, especially the hydrophilic compound with lower surface activity. In summary, these results suggest that an ideal ESI(-) modifier should provide cations that can be easily electrochemically reduced and produce anions with small molecular volume and high gas-phase proton affinity. PMID- 14750884 TI - A modified multisite Stern-Volmer equation for the determination of conditional stability constants and ligand concentrations of soil fulvic acid with metal ions. AB - In this work, we modify the multisite Stern-Volmer (MSV) equation for fitting fluorescence titration curves. Under the condition of a static quenching mechanism, the MSV postulates an underlying 1:1 fulvic acid (FA)/copper coordination ratio at multisites. Approximates of six fitting parameters characterize the stability constants (K1 and K2) of FA ligands with Cu2+, micromolar ligand site concentrations (CL1 and CL2), the unquenched, steady-state fractional fluorescence contributions (fx), and the residual fluorescence intensity (IRES). Prior to its application to actual FA titration data, the MSV function is simulated, and its predictive ability is confirmed by titrating a mixture of model fluorophores, glycyl-L-tryptophan and L-tryptophan with Cu2+ at pH 6. Molecular fluorescence measurements of FA are acquired at a fixed spectral position (lambda(ex) = 335 nm; lambda(em) = 450 nm), and FA is titrated with copper in triplicate at three pH values-5, 6, and 7. An objective analysis of log K(1) and K(2) values supports several site organization schemes, including (i) subtle, cooperative interaction, (ii) interfering molecular conformations, and (iii) aggregate forms. Site densities (CL1 and CL2) are consistent across varied pH. The f(1) is indicative of a pH-induced spectral shift of a fluorophore and convincingly associates with a transect in the Deltalambda = 25 synchronous fluorescence spectrum and with the preexponential terms describing the time dependent fluorescence decay. The MSV and its parent one-site version are equivalent for data fitting but are only simple approximations of a FA ligand system with more complex molecular fundamentals. PMID- 14750885 TI - Liquid-phase microextraction as an on-line preconcentration method in capillary electrophoresis. AB - A simple and efficient sample preconcentration method for capillary electrophoresis has been developed using liquid-phase microextraction (LPME). A thin layer of an organic liquid was used to separate a drop of the aqueous acceptor phase hanging at the inlet of a capillary from the bulk aqueous donor phase. The donor-phase pH was 1.0, and the acceptor phase pH was 9.5. This pH difference caused the preconcentration of the acidic compounds, fluorescein and fluorescein isothiocyanate, into the acceptor-phase drop. Enrichment factors of 3 orders of magnitude were obtained with 30-min LPME at 35 degrees C. PMID- 14750886 TI - Voltammetric sensor for general purpose organohalide detection at picogram per liter concentrations based on a simple collector-generator method. AB - With the aim of producing a general purpose sensor for environmental analysis, we describe a simple and sensitive method for organohalide detection, based on an electrochemical collector-generator process. The sensor consists of four coplanar electrodes contacting a solution volume of 300 microL, containing organohalide. At the first working electrode (a Zn/PTFE composite), the analyte is electrolyzed to liberate halide ions. At the second working electrode (Ag), the halide ions are detected by cathodic stripping voltammetry. Using a preconcentration time of 600 s, with differential pulse voltammetry for stripping, the responses to 1 chloropropane, chloroform, carbon tetrachloride, iodoethane, and bromoethane can be plotted on a common calibration curve, with a detection limit of 0.1 nM (1.3 pg L(-1) or less depending on the organohalide). To the best of our knowledge, this is the lowest reported organohalide detection limit by an electrochemical method and is so far the only general purpose electrochemical method sensitive enough for regulatory requirements. The sensor response was invariant for approximately 40 measurements. Analysis of tap water, spiked with chloroform or carbon tetrachloride, gave recoveries within 1.0-2.6% of the recoveries by the standard GC method. PMID- 14750887 TI - The Vinland Map ink is NOT medieval. PMID- 14750888 TI - Polarized reflectance spectroscopy for pre-cancer detection. AB - Early detection of cancer and its curable precursors remains the best way to ensure patient survival and quality of life. Thus, highly selective, sensitive and cost-effective screening and diagnostic techniques to identify curable pre cancerous lesions are desperately needed. Precancers are characterized by increased nuclear size, increased nuclear/cytoplasmic ratio, hyperchromasia and pleomorphism, which currently can only be assessed through an invasive, painful biopsy. Here, we describe the development of a non-invasive optical technique based on polarized reflectance spectroscopy that has the potential to provide in real time diagnostically useful information for pre-cancer detection. Our results demonstrate that polarized reflectance spectroscopy can be used to selectively detect the size-dependent scattering characteristics of nuclei in vivo. We gradually progress from cell suspensions to realistic three-dimensional tissue models of epithelium, then to cervical biopsies and, finally to in vivo studies on normal volunteers and clinical patients. PMID- 14750889 TI - Fluorescence imaging in vivo: raster scanned point-source imaging provides more accurate quantification than broad beam geometries. AB - Two fluorescence imaging systems were compared for their ability to quantify mean fluorescence intensity from surface-weighted imaging of tissue. A broad beam CCD camera system was compared to a point sampling system that raster scans to create the image. The effects of absorption and scattering in the background tissue volume were shown to be similar in their effect upon the signal, but the effect of the three-dimensional shape of the tissue was shown to be a significant distortion upon the signal. Spherical phantoms with Intralipid and blood for absorber and scatterer were used with a fixed concentration of aluminum phthalocyanine fluorophore to illustrate that the mean intensity observed with the broad beam system increased with size, while the mean intensity observed with the raster scanned system was not as significantly affected. Similar results were observed in vivo with mice injected with the fluorophore and imaged multiple times to observe the pharmacokinetics of the drug. The fluorescence in the tumor observed with the broad beam system was higher than that observed with the raster scanned system. Based upon the phantom and animal observations in this study, it should be concluded that using broad beam fluorescence imaging systems to quantify fluorescence in vivo may be problematic when comparing tissues with different three dimensional characteristics. In particular, the ratio of fluorescence from tumor to normal tissue can yield inaccurate results when the tumor is large. However, similar measurements with a narrow beam system that is raster scanned to create the images are not as significantly affected by the three dimensional shape of the tissue. Raster scanned imaging appears to provide a more uniform and accurate way to quantify fluorescence signals from distributed tissues in vivo. PMID- 14750890 TI - The imaging of apoptosis with the radiolabeled annexin V: optimal timing for clinical feasibility. AB - In recent years, the imaging of drug-induced apoptosis has become one of the centers of interest in experimental and clinical research. In particular, the accurate monitoring of chemosensitivity as well as the early prediction of chemoresistance in response to various pro-apoptotic interventions are critical requirements for the best management of oncology patients. The use of technetium [(99m)Tc]-labeled annexin V on animal and human models of cancers provides a proof of principle for the feasibility of a non-invasive, in vivo detection of an apoptotic signal and then for the early assessment of tumor response in the course of chemotherapy. Although promising, however, the initial clinical data point out on the technical limitations that are still to be resolved in terms of tumor-to-background ratio and optimal timing for the imaging of apoptosis. In the present review article, we report the results of animal studies aimed to the evaluation of apoptotic peaks following chemotherapy. In the light of these basic research works, we analyze the profiles of radiolabeled annexin V uptake over time as observed in clinical trials. We then discuss possible new imaging strategies designed to optimize the visualization of apoptotic changes within tumor tissues using the [(99m)Tc]-labeled annexin V. We also suggest longer lived forms of radiolabeled annexin V designed to better understand the temporal patterns of apoptotic tumor response, which in turn, may help to capture the best time-window for the imaging of cell death. PMID- 14750891 TI - Nanoshell-enabled photonics-based imaging and therapy of cancer. AB - Metal nanoshells are a novel type of composite spherical nanoparticle consisting of a dielectric core covered by a thin metallic shell which is typically gold. Nanoshells possess highly favorable optical and chemical properties for biomedical imaging and therapeutic applications. By varying the relative the dimensions of the core and the shell, the optical resonance of these nanoparticles can be precisely and systematically varied over a broad region ranging from the near-UV to the mid-infrared. This range includes the near infrared (NIR) wavelength region where tissue transmissivity peaks. In addition to spectral tunability, nanoshells offer other advantages over conventional organic dyes including improved optical properties and reduced susceptibility to chemical/thermal denaturation. Furthermore, the same conjugation protocols used to bind biomolecules to gold colloid are easily modified for nanoshells. In this article, we first review the synthesis of gold nanoshells and illustrate how the core/shell ratio and overall size of a nanoshell influences its scattering and absorption properties. We then describe several examples of nanoshell-based diagnostic and therapeutic approaches including the development of nanoshell bioconjugates for molecular imaging, the use of scattering nanoshells as contrast agents for optical coherence tomography (OCT), and the use of absorbing nanoshells in NIR thermal therapy of tumors. PMID- 14750892 TI - Stereotactic radiosurgery versus fractionated stereotactic radiotherapy boost for patients with glioblastoma multiforme. AB - The aim of this study is to evaluate the efficacy of stereotactic radiotherapy boost (SRB) in patients with glioblastoma multiforme (GBM) by comparing two different regimens, single dose or fractionated treatment. Between December 1994 and January 2000, 24 patients with GBM were treated with SRB in conjunction with external beam radiotherapy (EBRT). Fourteen patients (58%) were treated with stereotactic radiosurgery (SRS) and 10 patients (42%) with fractionated stereotactic radiotherapy (FSRT). Median interval between EBRT and SRS or FSRT was 1.4 months (range -0.4-3.9 months). Actuarial survival rates of the entire 24 patients at one and two years following SRB were 63% and 34% respectively, with median survival time of 16 months. Variables predicting survival were age, extent of surgery, re-operation and the RTOG (Radiation Therapy Oncology Group) classes based on recursive partitioning analysis (RPA). In comparison to historical controls, improved survival benefit after SRB was observed. The median survival times for the RTOG classes 4, 5, and 6 were 28.3, 10.3, and 6.0 months following EBRT+SRB, respectively. Expected values for these classes after EBRT are 11.1, 8.9, and 4.6 months, respectively. This improvement in survival was seen predominantly for the RTOG class 4. There was no difference in survival between SRS and FSRT treated groups. Late complications developed in 4 patients in the SRS group and 1 patients in the FSRT group. Our retrospective data suggest that SRB in conjunction with EBRT may improve survival in patients with GBM with median survival time of 16 months, when compared to historical controls of the RTOG data following EBRT. The addition of SRB appeared to improve the median survival most demonstrably in RTOG RPA class 4 patients. SRS and FSRT are equally effective with similar median survival, but potentially less late complications associated with FSRT. Since this is a nonrandomized study, further investigation is needed to confirm this and to determine an optimal dose/fractionation scheme. PMID- 14750893 TI - Enhanced surface registration techniques for patient positioning control in breast cancer radiotherapy. AB - Experimental data, describing patient inter-fractional set-up errors in the clinical practice of breast radiotherapy, were exploited to simulate the performance in errors detection and correction of a constrained surface registration procedure, based on a hybrid configuration of control points (passive and laser). During 47 treatment sessions in three patients undergoing post-quadrantectomy radiotherapy, an opto-electronic localizer was used to acquire the three-dimensional coordinates of the hybrid control points, being two passive markers placed on selected skin landmarks on the sternum. Laser scanning technique was also applied for the acquisition of the 3-D surface model of the irradiated body area, which was used as reference for the automatic position correction procedure. A constrained surface registration algorithm was applied to estimate the rigid spatial transformation, describing the local errors affecting the control points. The improvement of the irradiation geometrical set-up, by correcting the patient position according to the estimated spatial transformation parameters, was simulated. Results showed that the proposed surface registration method allowed us to detect and significantly (Wilcoxon signed rank analysis) reduce the initial misalignments, which exhibited overall median and 75th percentile values equal to 4.26 mm and 5.76 mm. Simulated residual errors dropped down to median and 75th percentile values measuring 2.95 mm and 3.87 mm, respectively. These results confirmed the high potentiality of surface registration techniques for the opto-electronic automatic patient positioning control in breast cancer radiotherapy. PMID- 14750894 TI - Repositioning accuracy of a commercially available double-vacuum whole body immobilization system for stereotactic body radiation therapy. AB - We evaluated the repositioning accuracy of a commercially available stereotactic whole body immobilization system (BodyFIX, Medical Intelligence, Schwabmuenchen, Germany) in 36 patients treated by hypofractionated stereotactic body radiation therapy. CT data were acquired for positional control of patient and tumor before each fraction of the treatment course. Those control CT datasets were compared with the original treatment planning CT simulation and analyzed with respect to positional misalignment of bony patient anatomy, and the respective position of the treated small lung or liver lesions. We assessed the stereotactic coordinates of distinct bony anatomical landmarks in the original CT and each control dataset. In addition, the target isocenter was recorded in the planning CT simulation dataset. An iterative optimization algorithm was implemented, utilizing a root mean square scoring function to determine the best-fit orientation of subsequent sets of anatomical landmark measurements relative to the original treatment planning CT data set. This allowed for the calculation of the x, y and z-components of translation of the patient's body and the target's center-of-mass for each control CT study, as well as rotation about the principal room axes in the respective CT data sets. In addition to absolute patient/target translation, the total magnitude vector of patient and target misalignment was calculated. A clinical assessment determined whether or not the assigned planning target volume safety margins would have provided the desired target coverage. To this end, each control CT study was co-registered with the original treatment planning study using immobilization system related fiducial markers, and the computed isodose calculation was superimposed. In 109 control setup CT scans available for comparison with their respective treatment planning CT simulation study (2-5 per patient, median 3), anatomical landmark analysis revealed a mean bony landmark translation of -0.4 +/- 3.9 (mean +/- SD), -0.1 +/- 1.6 and 0.3 +/- 3.6 mm in x, y and z-directions, respectively. Bony landmark setup deviations along one or more principal axis larger than 5 mm were observed in 32 control CT studies (29.4%). Body rotations about the x-, y- and z-axis were 0.9 +/- 0.7, 0.8 +/- 0.7 and 1.8 +/- 1.6 degrees, respectively. Assuming a rigid body relationship of target and bony anatomy, the mean computed absolute target translation was 2.9 +/- 3.3, 2.3 +/- 2.5 and 3.2 +/- 2.7 mm in x, y and z-directions, respectively. The median and mean magnitude vector of target isocenter displacement was computed to be 4.9 mm, and 5.7 +/- 3.7 mm. Clinical assessment of PTV/target volume coverage revealed 72 (66.1%), 23 (21.1%), and 14 (12.8%), of excellent (100% isodose coverage), good (>90% isodose coverage), and poor GTV/isodose alignment quality (less than 90% isodose coverage to some aspect of the GTV), respectively. Loss of target volume dose coverage was correlated with translations >5 mm along one or more axes (p<0.0001), rotations >3 degrees about the z-axis (p=0.0007) and body mass index >30 (p<0.0001). The analyzed BodyFIX whole body immobilization system performed favorably compared with other stereotactic body immobilization systems for which peer-reviewed repositioning data exist. While the measured variability in patient and target setup provided clinically acceptable setup accuracy in the vast majority of cases, larger setup deviations were occasional observed. Such deviations constitute a potential for partial target underdosing warranting, in our opinion, a pre-delivery positional assessment procedure (e.g., pre-treatment control CT scan). PMID- 14750895 TI - Inversely planned intensity modulated radiotherapy of the breast including the internal mammary chain: a plan comparison study. AB - The aim of this paper is to evaluate the benefit of inversely planned intensity modulated radiotherapy (IMRT) in the adjuvant irradiation of breast cancer when internal mammary lymph nodes are included in the treatment volume. 20 patients treated with 3D-planned conventional radiotherapy (CRT) following breast conserving surgery were included in the study. We chose 10 patients with left sided and 10 patients with right-sided tumors. All treatment volumes included the internal mammary chain. For plan comparison to the applied CRT plan an inverse IMRT-plan in, step-and-shoot'-technique was calculated. For all patients IMRT resulted in an improved conformity of dose distribution to the target volume compared to CRT (mean COIN95: 0.798 vs. 0.514 with COIN95 = C1 * C2 (C1= fraction of CTV that is covered by > 95% of the prescribed dose and C2 = volume of CTV that is covered by > 95% of the prescribed dose/total volume that is covered by > 95% of the prescribed dose). In all cases with matching adjacent beams, the homogeneity in the target volume was improved. The volume of the ipsilateral lung irradiated with a dose higher than 20 Gy was reduced with IMRT from 24.6% to 13.1% compared to CRT. For left-sided target volume the heart volume with a dose higher than 30 Gy was reduced from 6.2% to 0.2%. The presented plan comparison study for irradiation of the breast and the parasternal lymph nodes showed a substantial improvement of the dose distribution by inversely planned IMRT compared to CRT. This is visible for the target volume, the ipsilateral lung and, in case of left-sided target volume, the heart. Despite an increase in integral dose to the entire normal tissue, the application of IMRT might be clinically advantageous in cases where no satisfying dose distribution can be obtained by CRT. PMID- 14750896 TI - Improved display of synthetic IgG-binding domains on the baculovirus surface. AB - Improved display of foreign protein moieties in combination with beneficial alteration of the viral surface properties should be of value for targeted and enhanced gene delivery. Here, we describe a vector based on Autographa californica multiple nucleopolyhedrovirus (AcMNPV) displaying synthetic IgG binding domains (ZZ) of protein A fused to the transmembrane anchor of vesicular stomatitis virus (VSV) G protein. This display vector was equipped with a GFP/EGFP expression cassette enabling fluorescent detection in both insect and mammalian cells. The virus construct displayed the biologically active fusion protein efficiently and showed increased binding capacity to IgG. As the display is carried out using a membrane anchor of foreign origin, gp64 is left intact for virus entry, which may increase gene expression in the transduced mammalian cells. In addition, the viral vector can be targeted to any desired cell type via binding of ZZ domains when an appropriate IgG antibody is available. PMID- 14750897 TI - Liver tumor cryoablation: a commentary on the need of improved procedural monitoring. AB - Cryoablation is a method used for in situ destruction of liver tumors not eligible for surgical resection. Local recurrences following such treatment have been reported at rates of 5-44%. Insufficient procedural monitoring of the ablation is one plausible explanation for these recurrences. The cryoablative procedure is usually monitored by ultrasonography, but acoustic shadowing and loss of signals, compromise visualisation of the cryolesion circumference. Other monitoring modalities such as computer tomography and invasive methods like the use of thermocouples and impedance measurements have also been studied, but are not in common clinical use as single monitoring modalities. Thermodynamic conditions assumed adequate for tumor eradication are likely to occur only in parts of the cryolesion. This tumoricidal part of the cryolesion is not adequately depicted using any of these modalities. Magnetic resonance imaging (MRI) provides a clear delineation of the cryolesion circumference. Noninvasive temperature measurements assisted by MRI indicate which parts of the cryolesion that may be subject to complete necrosis. In this article MRI monitored cryoablation of liver tumors is discussed. Improved peroperative monitoring as offered by MRI may reduce the rates of local recurrences after treatment, but further technological improvements are required. PMID- 14750898 TI - Economic evaluations of granulocyte colony-stimulating factor: in the prevention and treatment of chemotherapy-induced neutropenia. AB - The prevailing uncertainty about the pharmacoeconomic positioning of granulocyte colony-stimulating factor (G-CSF) in the prevention and treatment of chemotherapy induced febrile neutropenia has resulted in a number of pharmacoeconomic evaluations published in the past 10 years. These studies vary considerably regarding the approaches used and the results presented. In order to contribute to a clearer pharmacoeconomic positioning of G-CSF, a systematic review of economic evaluations was carried out. The focus of the review was prophylaxis and therapy of chemotherapy-induced neutropenia in patients with cancer. A computerised bibliography search of several databases was conducted yielding 33 studies. The findings demonstrated the cost-saving potential of G-CSF in standard dose chemotherapy to be limited, with lower costs often seen in the control group. The results of these studies were too heterogeneous to extract a clear recommendation from a cost-saving point of view. The administration of G-CSF after high-dose chemotherapy with stem cell support resulted more often in cost savings in the G-CSF group as compared with standard-dose chemotherapy, illustrating a possible cost-saving potential of G-CSF. In the treatment of established chemotherapy-induced febrile neutropenia, cost savings were found in all studies. This result is surprising but hampered by the small number of studies (n = 5) and remains to be confirmed by more rigourously designed prospective economic analyses. Despite the substantial research on this topic, the economic evaluation of G-CSF is far from being settled and needs further investigation. PMID- 14750899 TI - The health economics of bladder cancer: a comprehensive review of the published literature. AB - The aim of this paper was to conduct a critical systematic review of the available literature on the clinical and economic burden of bladder cancer in developed countries, with a focus on the cost effectiveness of interventions aimed at reducing that burden.Forty-four economic studies were included in the review. Because of long- term survival and the need for lifelong routine monitoring and treatment, the cost per patient of bladder cancer from diagnosis to death is the highest of all cancers, ranging from 96000-187000 US dollars (2001 values) in the US. Overall, bladder cancer is the fifth most expensive cancer in terms of total medical care expenditures, accounting for almost 3.7 billion US dollars (2001 values) in direct costs in the US. Screening for bladder cancer in the general population is currently not recommended. The economic value of relatively new and less expensive urine assays and molecular urinary tumour markers has not been assessed. However, the literature suggests that screening patients suspected of having bladder cancer and using less invasive diagnostic procedures is cost effective. Very few cost-effectiveness studies have evaluated intravesical therapies such as bacillus Calmette-Guerin and mitomycin in the management of superficial disease and no robust recommendations can be drawn. Economic analyses suggest that non-surgical treatment strategies for the management of invasive disease aiming at bladder preservation may not be cost effective, because they have not consistently demonstrated survival benefits and do not eliminate the need for subsequent radical cystectomy. The literature suggests that the current conventional frequent follow-up and monitoring of patients can be cost effectively replaced by less frequent and less invasive monitoring, and should rely more heavily on intravesical chemotherapy to reduce the need for cystoscopies. Bladder cancer is a fairly common and costly malignancy. Nevertheless, the existing literature only contributes marginally to our knowledge concerning the burden of bladder cancer and the economic value of various interventions. The limited value of the literature in this area may be attributed to (i) being published as abstracts rather than full peer-reviewed evaluations; (ii) employing questionable methodologies; and (iii) being in many cases nearly obsolete, rendering them less relevant to, if not in conflict with, current clinical practice. Consequently, opportunities exist to conduct meaningful economic research in all areas of the management of bladder cancer, including screening, diagnosis, follow-up and treatment, especially with respect to new and innovative pharmaceutical and other technologies. PMID- 14750900 TI - An evaluation of the cost effectiveness of drotrecogin alfa (activated) relative to the number of organ system failures. AB - BACKGROUND: While drotrecogin alfa (activated) was shown to decrease absolute 28 day mortality by 6.1% in patients with severe sepsis in the Recombinant Human Protein C Worldwide Evaluation in Severe Sepsis (PROWESS) study, no mortality benefit was observed in the subset of patients with only one organ system failure. Consequently, some institutions restrict drotrecogin alfa (activated) use to patients with severe sepsis with >/=2 organ system failures. OBJECTIVE: To measure the cost effectiveness of drotrecogin alfa (activated) for treatment of severe sepsis in relation to the number of organ system failures and determine the economic impact of restricting drotrecogin alfa (activated) use based on the number of organ system failures. PERSPECTIVE: Policy perspective specific to our 340-bed, level I trauma centre. METHODS: A Monte Carlo simulation analysis was conducted to evaluate a hypothetical cohort of 10 000 patients with severe sepsis in four scenarios restricting treatment with drotrecogin alfa (activated) to patients with >/=1, >/=2, >/=3 or >/=4 organ system failures. The primary outcomes of 28-day all-cause mortality and serious bleeding were obtained from the PROWESS study. Costs (year 2002 values) were obtained from institutional financial records and literature estimates. The incremental cost per life saved at 28 days with drotrecogin alfa (activated) plus best standard care versus best standard care alone (placebo) was calculated. The incidence of severe sepsis and number of drotrecogin alfa (activated) candidates were estimated through chart review, and projected annual institutional expenditures were derived according to these data. RESULTS: With increasing number of organ system failures, the proportion of lives saved with drotrecogin alfa (activated) increased, and consequently the ICER decreased. Restriction of drotrecogin alfa (activated) to patients with >/=4 organ system failures was the most cost-effective scenario (0.11 lives saved; 56727 US dollars per life saved). For the nine patients that would be treated annually by our institution under this policy, one life would be saved at a total additional cost of 56160 US dollars per year. Use of the drug in patients with >/=1 or >/=2 organ system failures would save the greatest number of lives per year (4-5); however, restricting drotrecogin alfa (activated) to patients with >/=2 organ system failures would be the cheaper alternative (total additional cost 356022 US dollars vs 462204 US dollars . CONCLUSION: While restriction of drotrecogin alfa (activated) use to patients with sepsis with >/=4 organ system failures is the most cost-effective alternative, restriction to those with >/=2 organ system failures is the preferred alternative for our institution according to the number of lives saved and available financial resources. PMID- 14750901 TI - The anti-Mullerian hormone type II receptor: insights into the binding domains recognized by a monoclonal antibody and the natural ligand. AB - Anti-Mullerian hormone (AMH) [also called Mullerian inhibiting substance (MIS)] is a member of the transforming growth factor-beta family. AMH and its type II receptor (AMHR-II) are involved in the regression of the Mullerian ducts in the male embryo, and in gonadal functions in the adult. AMH is also known to be a marker of granulosa and Sertoli cell tumours. We selected a high-affinity monoclonal antibody, mAb 12G4, specific for human AMHR-II (hAMHR-II), by FACS analysis, Western blotting and immunohistochemical staining of a hAMHR-II transfected CHO (Chinese hamster ovary) cell line, normal adult testicular tissue and granulosa cell tumours. Using peptide array screening, we identified the binding sequences of mAb 12G4 and AMH on the receptor. Identification of Asp53 and Ala55 as critical residues in the DRAQVEM minimal epitopic sequence of mAb 12G4 definitively accounted for the lack of cross-reactivity with the murine receptor, in which there is a glycine residue in place of an aspartic acid residue. In a structural model, the AMH-binding interface was mapped to the concave side of hAMHR-II, whereas the mAb 12G4-binding site was located on the convex side. mAb 12G4, the first mAb to be raised against hAMHR-II, therefore has unique properties that could make it a valuable tool for the immunotargeting of tumours expressing this receptor. PMID- 14750902 TI - Repression of Smad4 transcriptional activity by SUMO modification. AB - Smad4 plays a key role in TGF-beta (transforming growth factor beta)/Smad mediated transcriptional responses. We show that Smad4 is sumoylated both in vivo and in vitro. Recent studies showed that sumoylation of Smad4 regulated its stability, but the effect of sumoylation on the intrinsic transcriptional activity of Smad4 was not defined. We show that overexpression of SUMO (small ubiquitin-related modifier)-1 and Ubc9 can inhibit a TGF-beta-responsive reporter gene, whereas co-transfection with SUMO-1 protease-1 (SuPr-1) can increase the TGF-beta response. We show further that mutation of the Smad4 sumoylation sites or co-transfection with SuPr-1 greatly increases Smad4 transcriptional activity. Moreover, direct fusion of SUMO-1 to the sumoylation mutant Smad4 potently inhibits its transcriptional activity. Thus, as it is being rapidly discovered that sumoylation inhibits the activities of many transcription factors, sumoylation also represses Smad4 transcriptional activity. The net effect of sumoylation of Smad4 can therefore be either stimulatory or inhibitory, depending on the target promoter that is analysed. PMID- 14750903 TI - Identification and characterization of plant glycerophosphodiester phosphodiesterase. AB - GPX-PDE (glycerophosphodiester phosphodiesterase; EC 3.1.4.46) is a relatively poorly characterized enzyme that catalyses the hydrolysis of various glycerophosphodiesters (glycerophosphocholine, glycerophosphoethanolamine, glycerophosphoglycerol, glycerophosphoserine and bis-glycerophosphoglycerol), releasing sn-glycerol 3-phosphate and the corresponding alcohol. In a previous study, we demonstrated the existence of a novel GPX-PDE in the cell walls and vacuoles of plant cells. Since no GPX-PDE had been identified in any plant organism, the purification of GPX-PDE from carrot cell walls was attempted. After extraction of cell wall proteins from carrot cell suspension cultures with CaCl2, GPX-PDE was purified up to 2700-fold using, successively, ammonium sulphate precipitation, gel filtration and concanavalin A-Sepharose. Internal sequence analysis of a 55 kDa protein identified in the extract following 2700-fold purification revealed strong similarity to the primary sequence of GLPQ, a bacterial GPX-PDE. To confirm the identity of plant GPX-PDE, an Arabidopsis thaliana cDNA similar to that encoding the bacterial GPX-PDE was cloned and overexpressed in a bacterial expression system, and was used to raise antibodies against the putative Arabidopsis thaliana GPX-PDE. Immunochemical assays performed on carrot cell wall proteins extracted by CaCl2 treatment showed a strong correlation between GPX-PDE activity and detection of the 55 kDa protein, validating the identity of the plant GPX-PDE. Finally, various properties of the purified enzyme were investigated. GPX-PDE is a multimeric enzyme, specific for glycerophosphodiesters, exhibiting a K(m) of 36 microM for glycerophosphocholine and active within a wide pH range (from 4 to 10). Since these properties are similar to those of GLPQ, the bacterial GPX-PDE, the similarities between plant and bacterial enzymes are also discussed. PMID- 14750905 TI - Low back pain, health status, and quality of life in older adults: challenge and opportunity. PMID- 14750906 TI - Strategic planning, operations management, and fiscal analysis and control: organizational imperatives. PMID- 14750907 TI - How does low back pain impact physical function in independent, well-functioning older adults? Evidence from the Health ABC Cohort and implications for the future. AB - OBJECTIVE: To determine the relationships between low back pain (LBP) frequency and intensity and self-reported and performance-based physical function in a large cohort of well-functioning older adults. DESIGN: Cross-sectional survey and examination. SETTING: Community-based cohort of the Health, Aging, and Body Composition (Health ABC) study. PARTICIPANTS: Participants were 2,766 community dwelling adults, aged 70-79; 42% were African American, 52% were men. OUTCOME MEASURES: 1) Back pain-location, frequency, intensity; 2) Hip and/or knee pain; 3) Body mass index (BMI); 4) Self-reported difficulty doing functional tasks; 5) Lower extremity function, using the battery from the Established Populations for Epidemiologic Studies in the Elderly (EPESE); 6) Self-rated health; 7) Comorbidity; 8) Depressive symptoms, using the Center for Epidemiological Studies Depression (CES-D) scale. RESULTS: LBP was common (36%), and its frequency/intensity was significantly associated with other pain and comorbidities. In gender-specific models, LBP frequency/intensity was not significantly associated with EPESE performance score after adjusting for age, race, BMI, CES-D score, knee pain, hip pain, and other comorbidities. LBP frequency/intensity, however, was significantly associated with self-reported difficulty with most functional tasks after adjusting for important confounders. CONCLUSIONS: Among well-functioning community-dwelling older adults, LBP frequency/intensity was associated with perceived difficulty in performing important functional tasks, but not with observed physical performance. The demonstrated dose-response relationship between pain frequency/intensity and self reported task performance difficulty underscores the importance of clinical efforts to treat pain without necessarily eradicating it. Additional work is needed to determine whether back pain is associated with a risk for progressive functional decline and loss of independence in older adults and whether therapeutic interventions can ameliorate decline and, therefore, preserve independence. PMID- 14750904 TI - Phosphorylation of clock protein PER1 regulates its circadian degradation in normal human fibroblasts. AB - Recent advances suggest that the molecular components of the circadian clock generate a self-sustaining transcriptional-translational feedback loop with a period of approx. 24 h. The precise expression profiles of human clock genes and their products have not been elucidated. We cloned human clock genes, including per1, per2, per3, cry2 and clock, and evaluated their circadian mRNA expression profiles in WI-38 fibroblasts stimulated with serum. Transcripts of hPer1, hPer2, hPer3, hBMAL1 and hCry2 (where h is human) underwent circadian oscillation. Serum stimulation also caused daily oscillations of hPER1 protein and the apparent molecular mass of hPER1 changed. Inhibitor studies indicated that the CKI (casein kinase I) family, including CKIepsilon and CKIdelta, phosphorylated hPER1 and increased the apparent molecular mass of hPER1. The inhibition of hPER1 phosphorylation by CKI-7 [ N -(2-aminoethyl)-5-chloro-isoquinoline-8 sulphonamide], a CKI inhibitor, disturbed hPER1 degradation, delayed the nuclear entry of hPER1 and allowed it to persist for longer in the nucleus. Furthermore, proteasome inhibitors specifically blocked hPER1 degradation. However leptomycin B, an inhibitor of nuclear export, did not alter the degradation state of hPER1 protein. These findings indicate that circadian hPER1 degradation through a proteasomal pathway can be regulated through phosphorylation by CKI, but not by subcellular localization. PMID- 14750909 TI - Chronic pain, depression, and quality of life: correlations and predictive value of the SF-36. AB - OBJECTIVE: Depression is a major barrier to effective pain relief. The SF-36 Health Survey may be useful as an outcome measure for chronic pain patients with and without depression. The study purpose was to determine the correlation between the SF-36 Mental Composite Scale t-score and depression type in chronic pain patients and the positive predictive value of the SF-36 in classifying depression type in chronic pain patients. DESIGN: Cross-sectional survey of chronic pain patients at baseline assessment. SETTING: Interdisciplinary pain management center in the North Central United States. PATIENTS: Two hundred forty two consecutive, chronic noncancer pain patients. INTERVENTIONS: Assessments of patient-reported health-related quality of life (SF-36), pain, pain type and diagnoses, mental health diagnoses, and patient demographics. OUTCOME MEASURES: SF-36 Health Survey, pain diagnoses, and depression diagnoses. RESULTS: Study participants were 160 women (66%) and 82 men (34%), 95% Caucasian, whose mean age was 46 (+/-0.8 SD) years (range: 19-83). All types and diagnoses of chronic pain were represented. The prevalence of major depressive disorder was 52%. The type of depression was highly correlated with SF-36 score (r=-0.567; P < 0.001). All chronic pain patients had very low SF-36 scores. Compared with U.S. population norms, chronic pain patients with and without depression had significantly lower SF-36 scores as measured by z-scores. Chronic pain patients with major depressive disorder had a significantly lower Mental Composite Score t-score than those with minor or no depression-34.1 and 47.6, respectively (P < 0.001). The positive predictive value of the SF-36 for differentiating major depression from minor or no depression was 98% (sensitivity=84.4%, specificity=93.9%). CONCLUSIONS: The SF 36 Mental Composite Score and all subscales were highly correlated with depression type in chronic pain patients. The positive predictive value of the SF 36 in classifying depression type was high. The SF-36 may be a useful clinical tool to measure health-related quality of life in chronic pain patients. In addition, the SF-36 was able to detect major depression and demonstrate a dose effect relationship between depression type (severity) and health-related quality of life in chronic pain patients. PMID- 14750908 TI - Lidocaine patch 5% with systemic analgesics such as gabapentin: a rational polypharmacy approach for the treatment of chronic pain. AB - OBJECTIVE: To assess the effectiveness and safety of the lidocaine patch 5%, a targeted peripheral analgesic, in the treatment of postherpetic neuralgia, painful diabetic neuropathy, and low back pain patients with incomplete responses to their current analgesic treatment regimen containing gabapentin. DESIGN: This was a 2-week, open-label, nonrandomized, multicenter pilot trial in the clinical setting. Patients with postherpetic neuralgia, painful diabetic neuropathy, or low back pain with partial responses (average daily pain intensity >4/10) to their current analgesic treatment regimen were included. Treatment consisted of daily application of up to four lidocaine patches to areas of maximal peripheral pain. Effectiveness was evaluated using the Brief Pain Inventory (BPI). Safety was assessed by adverse events, physical and neurologic examinations, vital signs, and clinical laboratory tests. RESULTS: Significant improvements in BPI measures of pain intensity and pain relief were reported for all groups of patients after 2 weeks of lidocaine patch 5% treatment. Significant improvements in BPI measures of pain interference with general activity, mood, walking ability, normal work, relationships with others, sleep, and enjoyment of life were noted. The lidocaine patch 5% was found to be safe and well tolerated. CONCLUSIONS: Results of this study highlight the potential advantages achieved with rational polypharmacy using a targeted peripheral analgesic, the lidocaine patch 5%, with centrally acting agents such as the anticonvulsant gabapentin. Controlled trials are warranted to further define the impact of such combination therapy. PMID- 14750910 TI - A survey of chronic noncancer pain patients prescribed opioid analgesics. AB - OBJECTIVES: Opioid analgesic drugs are sometimes advocated for chronic noncancer pain (CNCP). However, due to the paucity of studies assessing problematic opioid drug use in this population, evidence for such is inconclusive, and this issue remains controversial. This survey assessed problematic drug use among CNCP patients. PATIENTS/SETTING: Patients (N=104) prescribed opioids (mean duration of treatment 14.1 months) for severe CNCP at a pain clinic within a National Health Service hospital in London, United Kingdom. DESIGN: A review of pain clinic records to identify CNCP patients who had been prescribed opioids and subsequent assessment of those patients for problematic drug use using a substance use questionnaire. RESULTS: A total of 90 (86.5%) patients reported stopping opioid therapy at some point and, of these, 59 (65%) had ceased opioid therapy permanently. Of those patients who stopped opioids, 13 reported opioid withdrawal symptoms, two with severe and two with very severe symptoms. However, 72.5% of all patients derived benefit from opioids, although 77% of all patients reported opioid side effects. The addiction rate was 2.8%. CONCLUSION: These findings indicate that opioid therapy for CNCP does not necessarily lead to problematic drug use. Some problematic side effects are likely to be surmountable through appropriate prescribing. Further research is required into the long-term use of opioids in CNCP. PMID- 14750911 TI - Menstrual pain intensity, coping, and disability: the role of pain catastrophizing. AB - OBJECTIVE: Menstrual pain or primary dysmenorrhea has not received much attention in the field of pain research. Little is understood about the effects menstrual pain has on the women who experience it. No studies to date have examined the cognitive factors related to the perceived intensity and coping of menstrual pain. To investigate these areas further, this study examined the associations between pain catastrophizing and how women perceive and cope with menstrual pain. DESIGN: A prospective and retrospective between-subjects study. PARTICIPANTS: Ninety-three undergraduate women, with a regular menstrual period and no preexisting pain disorder (e.g., endometriosis) that affects menstrual pain, were classified into high or low pain catastrophizing groups. OUTCOME MEASURES: Participants completed several self-reported questionnaires assessing pain catastrophizing, menstrual pain intensity, coping, and disability. RESULTS: High pain catastrophizers, in comparison with low pain catastrophizers, reported greater menstrual pain intensities, greater affective menstrual pain intensity, greater variability in the use of pain coping strategies, lower perceived effectiveness of over-the-counter medications and nonmedical pain coping strategies, and greater disability. CONCLUSIONS: The results extend our knowledge about the associations between pain catastrophizing and menstrual pain, reemphasize that pain experience is best viewed as a multidimensional construct, and have implications for the management of menstrual pain. PMID- 14750912 TI - On being a legal expert. Opinion No.1. PMID- 14750913 TI - On being a legal expert. Opinion No.2. PMID- 14750914 TI - On being a legal expert. Opinion No.3. PMID- 14750915 TI - On being a legal expert. Opinion No.4. PMID- 14750916 TI - A placebo for the pain: a medico-legal case analysis. AB - OBJECTIVES: The objectives of this medico-legal case report are to consider the current status of the use of placebos in pain medicine from clinical, ethical, and legal perspectives. The focus of the analysis is a particular case in which the deceptive use of placebo pain therapy on an adolescent gave rise to professional grievances filed by the patient's mother against the physician who ordered and several nurses who administered the placebo. The medical board declined to take disciplinary action against the physician, and disciplinary action by the board of registered nursing against the nurses was successfully challenged by two of the charged nurses in an administrative review. While there is a growing literature that challenges the need for or justification of the deceptive use of placebos, the practice continues and, as the case under consideration indicates, retains some influential supporters. DESIGN: This is a case report from a community hospital. The patient, referred to here as KC, was an adolescent with migraine headaches. The substitution of a placebo (saline solution) for an opioid analgesic (morphine) was made during KC's treatment. RESULTS: The patient's pain subsided sufficiently following the administration of a placebo to permit his discharge from the hospital. The subsequent discovery by the patient's mother of the deceptive use of a placebo prompted her to file charges of professional misconduct against the treating physician and three nurses with their respective professional licensing boards. The medical board declined to take disciplinary action against the physician, and the disciplinary action by the board of registered nursing was successfully challenged in a ruling by an administrative law judge following a hearing in which expert witnesses took conflicting positions on the acceptability of the deceptive use of a placebo. CONCLUSION: While there is a developing literature that challenges the ethical legitimacy of the deceptive use of placebos in pain medicine, that literature has yet to be recognized as unqualifiedly setting the standard of care or of professionalism in medicine and nursing. PMID- 14750917 TI - Successful treatment of spontaneous cerebrospinal fluid leak headache with fluoroscopically guided epidural blood patch: a report of four cases. AB - Spontaneous cerebrospinal fluid (CSF) leak is a rare clinical entity that may result in disabling headaches. It occurs as a result of dural defects, and the initial symptoms resemble those of postdural puncture headache. However, the positional headache can later evolve into a persistent chronic daily headache. The diagnosis of spontaneous CSF leak can be very challenging, but increasing awareness and improved diagnostic techniques are yielding ever more cases. When conservative management fails, the pain management clinician is called upon to administer an epidural blood patch. The success of this technique is dependent upon accurate diagnosis of the site of leakage and targeted epidural administration of the blood patch to this area. In this report, we describe four consecutive cases that were referred to our pain management department over an 18 month period and were successfully treated with site-directed epidural blood patches. PMID- 14750918 TI - Improvement in sexual function after reduction of chronic high-dose opioid medication in a cancer survivor. AB - OBJECTIVE: To demonstrate improvement in sexual function after reduction of opioids. METHODS: This was a retrospective examination of a single patient at the cancer pain management clinic at M.D. Anderson Cancer Center in Houston, Texas. The patient was a 58-year-old male, free of cancer for 12 years, with chronic low back pain from a prior retroperitoneal mass. Changes in scores from the Brief Male Sexual Inventory and visual analog scale pain questionnaires were used to evaluate the patient. RESULTS: In this patient, a decrease in morphine-equivalent daily dose from 690 mg to 20 mg resulted in a significant increase in sexual function. Sexual inventory scores increased from 4 to 43. CONCLUSIONS: Reduction in opioid consumption can dramatically increase libido and sexual function. A possible mechanism involves opioid-related effects on the hypothalamic-pituitary gonadal axis. PMID- 14750919 TI - Waddell signs. PMID- 14750921 TI - Why withholding a prescription is unethical: a bioethical analysis of a far too familiar case. PMID- 14750922 TI - Working at the World Health Organization: An international perspective on mental health nursing. PMID- 14750923 TI - Valuing the past: The importance of an understanding of the history of psychiatry for healthcare professionals, service users and carers. AB - Drawing primarily on examples from the UK, this paper argues that an under standing of the history of psychiatry can be of practical use to clinicians, service users and carers. It can provide a window on the present and an explanation of some aspects of current practice. In addition, a study of the past can provide a vision of how things might be done differently in the present and in the future. Moreover, knowledge of the past can also serve to remind us of psychiatry's potential for misuse and the dangers inherent in a view of the world in which psychiatry is simply seen as a benevolent science destined to find an answer to mental illness. PMID- 14750924 TI - Rural community mental health nursing: a grounded theory account of sole practice. AB - This paper examines community mental health in rural and remote settings, characterized as sole practice. Using a grounded theory approach, the research reported here explored how meanings of health and health care are advanced within the context of rural mental health care, dominated and in the main led by nurses. Five different practice sites in rural New South Wales were involved. The study articulated a model of therapeutics that foregrounds a relationship of intense professional intimacy and trust against a context of geographical disadvantage and professional isolation. The meanings of the relationship are elaborated in terms of unusually high levels of responsibility, professional ingenuity, powerlessness and the independent and risky character of life in the bush. PMID- 14750925 TI - Implementing research findings into mental health nursing practice: exploring the clinical research fellowship approach. AB - The lack of research utilization within nursing practice has been extensively discussed in the literature. The Clinical Research Fellowship (CRF) program was developed to assist nurses to change practice on the basis of high-quality research evidence. This paper presents the results of a qualitative study examining the experiences of four CRF participants and three of their unit managers in completing the program and implementing changes within the clinical setting. The major themes to emerge from the data were: experience of the program, outcomes, implementation, assistance from the Centre for Psychiatric Nursing Research and Practice, Victoria, Australia, benefits and drawbacks to the program and whether it would be recommended to others. The findings indicate a positive view of the program itself although problems with the implementation stage were clearly evident. Further support following completion of the program is required to achieve maximum benefit from the program. PMID- 14750926 TI - The development of a professional practice audit questionnaire for mental health nursing in Aotearoa/New Zealand. AB - This paper reports the three-stage development of a professional practice audit questionnaire for mental health nursing in Aotearoa/New Zealand. In Study 1, clinical indicator statements (n = 99) generated from focus group data, which were considered to be unobservable in the nursing documentation in consumer case notes, were included in a three-round Delphi process. Consensus of ratings occurred for the mental health nurse and academic participants (n = 7) on 83 clinical indicator statements. In Study 2, the clinical indicator statements (n = 67) that met importance and consensus criteria were incorporated into a questionnaire, which was piloted at a New Zealand mental health service. The questionnaire was then modified for use in a national field study. In Study 3, the national field study, registered mental health nurses (n = 422) from 11 New Zealand District Health Board mental health services completed the questionnaire. Five categories of nursing practice were identified: professional and evidence based practice; consumer focus and reflective practice; professional development and integration; ethically and legally safe practice; and culturally safe practice. Analyses revealed little difference in the perceptions of nurses from different backgrounds regarding the regularity of the nursing practices. Further research is needed to calibrate the scores on each clinical indicator statement with behaviour in clinical practice. PMID- 14750927 TI - Hassles and uplifts associated with caring for people with cognitive impairment in community settings. AB - In this study we explored the hassles and uplifts (i.e. negative and positive emotional events) experienced by registered nurses, nursing assistants and personal carers working with people with cognitive impairment in community and residential healthcare settings in Brisbane, Queensland, Australia. The primary aim of the research was to explore what aspects of caring for cognitively impaired clients hassles nurses, what helps to relieve these hassles, what aspects of this work nurses find rewarding and what detracts from those rewards, as well as the intensity with which each of these aspects were felt. A questionnaire developed to explore hassles and uplifts at work was administered and 57 responses obtained. Results indicated that caring for the cognitively impaired client provides many uplifts for nurses and few hassles. However, the hassles that occurred were of high importance. This paper will be of interest to managers, nurses and carers in settings where there are people with cognitive impairment as well as scholars, who may find that assessing emotional hassles and uplifts provides additional insights into other areas of nursing. PMID- 14750928 TI - A grounded theory study of the role that nurses play in increasing clients' willingness to access community mental health services. AB - Clients and caregivers frequently encounter difficulties in accessing mental health services. Early intervention to care and treatment is essential to recovery as delays can adversely affect the severity and the duration of the illness. This paper reports the factors that encouraged and deterred young adults with schizophrenia and their caregivers when trying to seek early access to community mental health services, and examines how community mental health nurses made themselves accessible to these individuals in order to increase their access to services. The findings showed that several factors simultaneously encouraged and inhibited individuals initiating contact at community mental health centres, and these had serious implications for care and treatment, and recovery. There were two client access pathways to care: a direct access pathway where an individual recognized signs of being unwell and sought help early; and an indirect access pathway where others, such as caregivers, general practitioners, police, and inpatient facilities initiated contact on an individual's behalf. Nurses used three strategies to enhance client and caregiver access to services: 'promoting favourable experiences to enhance approachability', 'using technology to promote access' and 'being available'. The findings have significance for nursing practice because they reinforce the importance of having good relationships with clients and caregivers, and the need to take account of the needs of caregivers. They highlight the benefits and drawbacks of using telecommunications to enhance accessibility, and why it is necessary to analyse nurses' workloads and time management skills. PMID- 14750929 TI - Measuring the effectiveness of the national Mental Health Triage Scale in an emergency department. AB - The mainstreaming process has significantly altered the means of access to mental health services in Australia. Increasingly people seeking mental health care present at general hospital emergency departments. The triage system, which has proven effective for prioritizing physical illness and injury, has proven problematic when applied to mental health-related problems. This paper presents the results of a study undertaken in the emergency department of a Victorian public hospital. The Mental Health Triage Scale was introduced and used independently by triage nurses and the psychiatric nurse consultants employed in the department. Following a 3-month period, the two sets of triage scores for psychiatric presentations (n = 137) were compared. The findings suggest that triage nurses are rating clients experiencing mental health problems as in more urgent need of care than their psychiatric nursing counterparts. This suggests that the introduction of the guidelines alone is insufficient, and that education is required for more effective use of the tool. PMID- 14750930 TI - The World Federation of Hemophilia: 40 years of improving haemophilia care worldwide. AB - In 2003, the World Federation of Hemophilia marks its 40th anniversary. Established in 1963 by Frank Schnabel, a person with hemophilia from Montreal, Canada, the WFH has grown into its role as an independent, not-for-profit representative of the global haemophilia community. Since then, its biannual world congresses have provided an opportunity to exchange information on research and treatment. Today, more than 3,000 delegates attend these congresses. Until the early 1990s, the WFH's other major function was the International Hemophilia Training Centre programme, offering training fellowships and workshops to medical and paramedical staff from developing countries. In 1982, AIDS was reported in people with haemophilia who had received tainted blood products. The WFH developed task forces and committees to monitor safety and supply issues and set up the Global Forum on the Safety and Supply of Treatment Products. In the mid 1990s, twinning programmes were established for treatment centres and national haemophilia organizations. Twinning facilitates the exchange of training, coaching, and expertise. In the late 1990s, the WFH expanded and began working with local organizations and health authorities in a number of countries to improve the diagnosis and care of persons with hemophilia. The federation currently has ongoing projects in 25 developing countries. On World Hemophilia Day, April 17, 2003, the WFH launched the Global Alliance for Progress (GAP) in haemophilia which aims to double the number of people with haemophilia diagnosed and receiving treatment in up to 40 developing countries over a 10-year period. PMID- 14750931 TI - Why is primary prophylaxis underutilized in the United States? AB - A recent article published in Haemophilia reports the results of a survey of treatment center practices in the United States, and specifically the use of prophylaxis as treatment regimen. This article discusses the issues that may contribute to utilization of prophylactic regimens and issues to consider when determining treatment programs for pediatric patients. PMID- 14750932 TI - The dental management of adult patients with haemophilia and other congenital bleeding disorders. AB - The management of adult dental patients with congenital bleeding disorders has caused a considerable number of problems to the dental profession. There is a need to simplify the process and identify what can be safely carried out on a 'shared care' basis in General Dental Practice or the Community Dental Service. Particular problems are discussed with special reference to those requiring hospital care. The Scottish Oral Health Group for Medically Compromised Patients has developed this clinical guidance in conjunction with the Scottish Haemophilia Directors. It is important that dental care is easily available for this group of patients, especially those living at some distance from the regional centres. The aim is to simplify planning dental care for this group of patients and remove a number of myths concerning their management. The hospital departments, both medical and dental, must be available for advice and to arrange for treatment that is inappropriate outside a specialist hospital. PMID- 14750933 TI - Health-related quality of life assessment in adult haemophilia patients: a systematic review and evaluation of instruments. AB - The objective of this study was to review and evaluate the performance of health related quality of life (HRQL) and other health status measures used in studies of adult haemophilia and provide recommendations for future research. A systematic literature review was performed to identify HRQL and health status measures used in haemophilia. Literature was identified using medical databases, Internet and manual searches. The search was restricted to articles published in English since 1986. Ninety-six abstracts were located; 19 relevant articles were selected for detailed review. Three main types of HRQL measures were identified: generic psychometric-based HRQL (SF-36 and SF-12), utility-based HRQL [EQ-5D and Health Utilities Index (HUI)], and musculoskeletal-specific HRQL (Arthritis Impact Measurement Scale 2, AIMS 2) instruments. No patient-rated haemophilia specific HRQL instrument was found. The SF-36, the EQ-5D, and the HUI were able to discriminate haemophilia patient subgroups with respect to disease severity and HIV comorbidity status. Sixteen additional scales were identified that were used to measure the different aspects of physical, psychological, and social functioning of patients. There were no clinical studies of haemophilia carried out that employed HRQL instruments, thus responsiveness of these instruments could not be evaluated. The variety of instruments used in haemophilia studies highlights the need for a tool that can capture the full impact of haemophilia and its treatment on patients' HRQL. Developing such a tool poses the unique challenge of accounting for common comorbidities, such as HIV and chronic hepatitis that may have a greater HRQL impact than the underlying disease. PMID- 14750934 TI - Successful treatment of urgent bleeding in von Willebrand disease with factor VIII/VWF concentrate (Humate-P): use of the ristocetin cofactor assay (VWF:RCo) to measure potency and to guide therapy. AB - This prospective, open-label, non-randomized study evaluated the safety and efficacy of factor VIII (FVIII)/von Willebrand Factor (VWF) concentrate (Humate P) using treatment regimens based on VWF:ristocetin cofactor (VWF:RCo) activity in patients with von Willebrand Disease (VWD) in (i) urgent bleeding episodes, or (ii) in patients undergoing urgent and necessary surgery. This article summarizes the results of treatment for the 33 patients with 53 urgent bleeding events. The median loading dose of FVIII/VWF concentrate was 67.0 international units per kilogram (IU kg(-1)) VWF:RCo (range 25.7-143.2 IU kg(-1)), and the median daily maintenance dose per infusion was 74.0 IU kg(-1) (range 16.4-182.9 IU kg(-1)) for a median duration of 2 days (range 1-34 days). The overall efficacy (achievement of haemostasis) of FVIII/VWF concentrate was rated as excellent/good for 98% of the urgent bleeding events. No unexpected treatment-related adverse events or serious drug-related adverse events (AEs) were observed. This study supports the safety and efficacy of Humate-P administered in doses calculated in VWF:RCo units for the treatment of urgent bleeding episodes in patients with VWD. PMID- 14750935 TI - Retrospective analysis of 1312 patients with haemophilia and related disorders in a single Chinese institute. AB - With 1.3 billion people, China has the largest population in the world, and therefore has the largest population of persons with haemophilia (PWH). As there is no national registry for haemophilia, it is difficult to ascertain how many PWH have actually been diagnosed. Between January 1983 and June 2002, 1312 patients with coagulation disorders were referred to our hospital, and 1190 patients were evaluable. Among them, 1069 (89.8%) patients had haemophilia, 68 had vWD, 20 had factor XI deficiency, 10 had acquired factor VIII inhibitor and 23 had other coagulation disorders. Of the 1069 PWH, 14.7% were unclassified, 38.4% severe, 35.7% moderate and 11.1% mild. If the unclassified cases were excluded, 45.1% were severe, 41.9% moderate and 13.0% mild. Twenty-nine of the 68 vWD patients had vWF:Ag <5%, and subcategorized as type 3 vWD. Because vWF multimer analysis was not performed in our centre, the remaining vWD patients were not subdivided. PMID- 14750936 TI - Renal disease among males with haemophilia. AB - Haematuria is common among persons with haemophilia (PWH), but its long-term effects on the kidney and renal function are not well defined. In addition, infection with human immunodeficiency virus (HIV) or hepatitis C, or exposure to nephrotoxic agents as therapy for these infections may place PWH at increased risk for renal disease. To examine factors associated with chronic renal disease (CRD) and acute renal disease (ARD) in PWH, we analysed data collected from the medical records of 3422 males with haemophilia living in six US states from 1993 to 1998. Renal disease cases were ascertained from among 2075 persons who were hospitalized at least once over the 6-year period. Of these, 60 (2.9%) were diagnosed during one or more hospitalizations with either ARD (29/60) or CRD (31/60). In multivariate analyses, we examined associations between renal disease and demographic and clinical factors including age, race, haemophilia type and severity, hypertension, diabetes, history of recent renal bleeds, presence of an inhibitor, and infection with hepatitis C or HIV. HIV infection and hypertension were strongly associated with both ARD and CRD. PWH who had ARD were also more likely to have an inhibitor than those without this diagnosis. PWH who had CRD were more likely to be older and non-white and to have had a recent admission for a kidney bleed than those without diagnosed CRD. In summary, we found that HIV infection and haemophilia-related factors including inhibitors and kidney bleeds were associated with renal disease in a cohort of males with haemophilia. PMID- 14750937 TI - Immunoadsorption for coagulation factor inhibitors: a retrospective critical appraisal of 10 consecutive cases from a single institution. AB - Immunoadsorption is occasionally used as an adjuvant measure in the treatment of subjects with coagulation factor inhibitors. We reviewed our recent 3-year period experience with this procedure in 10 subjects. Immunoadsorption was used in the context of an immune tolerance protocol for 3 subjects with severe congenital deficiency in factor VIII, IX, and XIII; it was effective in lowering the level of inhibitor but immune tolerance was not achieved. It allowed successful use of porcine factor VIII in 4 cases of acquired hemophilia and in one case of inhibitor in mild hemophilia A. This therapeutic approach seems to be more useful in acquired hemophilia than in severe congenital factor deficiencies with inhibitors. PMID- 14750938 TI - Germ-line origin of intron 1 inversion in two haemophilia A families. AB - Factor VIII gene inversion of intron 1 has recently been reported to be the mutation responsible for haemophilia A in about 5% of severe cases. In our series of patients, which is made up of 77 Italian cases negative for intron 22 inversion, the mutation was found in three sporadic and in one familial patients, with an overall frequency of 5.2%. The carrier status of the patients' female relatives was assessed by mutation analysis and showed that only two-thirds of cases could be considered truly sporadic. The germ-line origin of the mutation was investigated in the two sporadic families by haplotype analysis on genomic DNA of the patients' maternal grandparents. These studies indicated that both mutation events had occurred in the germ cell lines of the patients' healthy grandfather, suggesting that, as already demonstrated for the inversion of intron 22, the male germ cell line is more susceptible to the intrachromosome recombination which leads to the inversion of intron 1. PMID- 14750939 TI - Iliopsoas haemorrhage in patients with bleeding disorders--experience from one centre. AB - Iliopsoas haemorrhage in patients with bleeding disorders is a potentially life threatening condition, with significantly associated morbidity. Despite its clinical importance, little has been published on the frequency, complications or outcomes of this entity since the advent of modern therapies for haemophilia. In a retrospective review of 297 patients with bleeding disorders followed at our centre, we identified 46 episodes of iliopsoas haemorrhage in 31 patients. Patients presented primarily with thigh, hip and/or groin pain, and frequently had flexion hip contracture, femoral nerve paresthesia, and >2 g dL(-1) haemoglobin drop. The duration of symptoms prior to seeking medical attention was 3.8 +/- 4 days. Nineteen of 155 patients (12.3%) with haemophilia A had 28 episodes of iliopsoas bleed; 52.6% of these patients had severe haemophilia. Of these 19 patients with haemophilia A who had iliopsoas haemorrhage, seven (36.8%) had an inhibitor to factor VIII (FVIII), and accounted for one-half of the bleeding episodes. Nine of 66 patients (13.6%) with haemophilia B had 15 episodes of iliopsoas haemorrhage; 22.2% of these patients had severe haemophilia, including one patient with an inhibitor to FIX who had two iliopsoas bleeds. The mean duration of therapy was 18.7 +/- 11.9 days, and the duration of hospitalization was 12.3 +/- 9.1 days. The length of hospital stay was significantly longer in patients with inhibitors, when compared with patients without inhibitors (19.1 +/- 5.8 days vs. 7.6 +/- 7.8 days; P<0.0001) with higher factor consumption, although the total duration of therapy was not significantly different. Patients with inhibitors were over-represented in the cohort of haemophiliacs with iliopsoas bleed. Patients with inhibitors who had iliopsoas bleeds remained hospitalized longer, although the duration of therapy was the same as patients with no inhibitors. There was a low frequency of recurrent bleed (2.8%). PMID- 14750940 TI - Molecular challenges and viral diseases. PMID- 14750941 TI - Concurrent total hip and knee replacements in a patient with haemophilia with inhibitors using recombinant factor VIIa by continuous infusion. AB - The introduction of activated recombinant factor VIIa (rFVIIa) has allowed elective surgery to be safely performed in haemophiliacs with inhibitors. The main problems associated with its use are the short half-life, necessitating frequent intravenous injections, and its very high cost. Here we describe, for the first time, the performance of total hip and knee replacements in a haemophiliac with inhibitors at the same operation. The amount of rFVIIa concentrate used (8.57 mg) was similar to that normally used for a single joint replacement. The use of continuous infusion allowed for easier administration and further contributed to the reduction in cost as it avoids the peak levels associated with bolus injections. PMID- 14750942 TI - Successful treatment of acquired haemophilia with prednisolone therapy. AB - Acquired hemophilia is a rare, life threatening coagulopathy in adults caused by the development of autoantibodies against to factor VIII. No general consensus exists on the best therapeutic approach. We report here a case that presented with extensive cutaneous and mucosal bleedings due to factor VIII inhibitors and treated successfully with steroid therapy alone but complicated with a life threatening thromboembolic attack during her follow up. In conclusion, corticosteroids are "cost effective therapy" associated with high inhibitor elimination rates and although recurrence of inhibitor in a patient with factor VIII inhibitor is an expected clinical situation thrombosis risk should also be considered. PMID- 14750943 TI - Meta-analysis on the effectiveness of B-domain deleted factor VIII for prophylaxis. PMID- 14750944 TI - Comparative study of full-length and B-domain deleted factor VIII concentrates. PMID- 14750945 TI - Controversial use of meta-analysis for factor VIII pharmacokinetic data. PMID- 14750947 TI - Crystal structure of the Pyrococcus horikoshii DNA primase-UTP complex: implications for the mechanism of primer synthesis. AB - BACKGROUND: In chromosomal DNA replication, DNA primase initiates the synthesis of a dinucleotide on a single-stranded template DNA, and elongates it to form a primer RNA for the replicative DNA polymerase. Although the apo-structure of an archaeal primase has been reported, the mechanism of primer synthesis by the eukaryotic-type primase still remains to be elucidated. RESULTS: In this study, we present the crystal structure of the eukaryotic-type DNA primase from the hyperthermophilic archaeon (Pyrococcus horikoshii) with the uridine 5' triphosphate (UTP). In the present primase-UTP complex, the primase binds the triphosphate moiety of the UTP at the active site, which includes Asp95, Asp97, and Asp280, the essential residues for the nucleotidyl transfer reaction. CONCLUSION: The nucleotide binding geometry in this complex explains the previous biochemical analyses of the eukaryotic primase. Based on the complex structure, we constructed a model between the DNA primase and a primer/template DNA for the primer synthesis. This model facilitates the comprehension of the reported features of DNA primase. PMID- 14750948 TI - Prion domain interaction responsible for species discrimination in yeast [PSI+] transmission. AB - BACKGROUND: The yeast [PSI+] factor is transmitted by a prion mechanism involving self-propagating Sup35 aggregates. As with mammalian prions, a species barrier prevents prion transmission between yeast species. The N-terminal of Sup35 of Saccharomyces cerevisiae, necessary for [PSI+], contains two species-signature elements-a Gln/Asn-rich region (residues 1-41; designated NQ) that is followed by oligopeptide repeats (designated NR). RESULTS: In this study, we show that S. cerevisiae[PSI+] is transmissible through plasmid shuffling and cytoplasmic transfer to heterotypic Sup35s whose NQ is replaced with the S. cerevisiae NQ. In addition to homology, the N-terminal location is essential for NQ mediated susceptibility to [PSI+] transmission amongst heterotypic Sup35s. In vitro, a swap of NQ of S. cerevisiae Sup35 led to cross seeding of amyloid formation. CONCLUSIONS: These findings suggest that NQ discriminates self from non-self, and is sufficient to initiate [PSI+] transmission irrespective of whether NR is heterotypic. NR as well as NQ alone coalesces into existing [PSI+] aggregates, showing their independent potentials to interact with the identical sequence in the [PSI+] conformer. The role of NQ and NR in [PSI+] prion formation is discussed. PMID- 14750949 TI - Suppression of spontaneous and hydrogen peroxide-induced mutations by a MutT-type nucleotide pool sanitization enzyme, the Escherichia coli Orf135 protein. AB - BACKGROUND: We recently found that the Escherichia coli Orf135 protein, a MutT type enzyme, hydrolysed 2-hydroxy-dATP (2-OH-dATP), and less efficiently, 8 hydroxy-dGTP. RESULTS: In this study, we examined the effects of the absence of the orf135 gene. Frequencies of spontaneous and H2O2-induced mutations were two- to three-fold higher in the orf135- strain than in the wild-type strain. These mutations include various mutations involving a G:C-->T:A transversion, the same type of mutation elicited by 2-OH-dATP. Over-expression of the Orf135 protein suppressed mutations even in the wild-type strain, as well as in the orf135- strain. CONCLUSIONS: The mutator phenotype of bacteria lacking the Orf135 protein suggests that this protein is involved in the suppression of mutations induced by oxidized deoxynucleotides in vivo and that various MutT-type enzymes contribute to nucleotide pool sanitization. PMID- 14750951 TI - Antioxidant function of a novel selenoprotein in Drosophila melanogaster. AB - BACKGROUND: Insects appear to have diverged from both higher and lower organisms in their defense mechanisms against oxidative damage. They do not encode glutathione peroxidases or glutathione reductases, and their thioredoxin reductases exhibit distinct properties from those of higher and lower species. Nonetheless, appropriate balance of anti-oxidants and pro-oxidants, and protection from damaging reactive oxygen species are clearly crucial in insects for viability, normal functioning of signalling pathways and morphogenesis, and have been implicated in studies on longevity in flies and other organisms. RESULTS: Two novel selenoproteins, dselH and dselK, were recently identified in Drosophila melanogaster. We have used RNAi in D. melanogaster embryos and in Schneider S2 cells to inhibit expression of these proteins. We report that inhibition of either dselH or dselK expression significantly reduces viability in embryos. We further show that dselH silencing decreases total anti-oxidant capacity in embryos and Schneider cells, and increases lipid peroxidation in cells. Conversely, transient expression of dselH in the cell line decreases lipid peroxidation, and reverses the toxic effects of a glutathione-depleting drug. The latter correlates with sparing of glutathione levels. CONCLUSIONS: These studies suggest that the well-known role of selenoproteins in vertebrate anti-oxidant defenses also extends to include invertebrates. PMID- 14750950 TI - Carboxy-terminal region of the yeast heat shock factor contains two domains that make transcription independent of the TFIIH protein kinase. AB - BACKGROUND: Phosphorylation of the carboxy-terminal domain (CTD) of the largest subunit of RNA polymerase II is implicated in transition from initiation to elongation in the transcription cycle. In yeast cells, Kin28, a subunit of the general transcription factor TFIIH, is responsible for the CTD phosphorylation. Although Kin28 is indispensable for transcription of many genes, its requirement is bypassed in certain genes such as SSA4 or CUP1, whose transcription is activated by the heat shock factor Hsf1. RESULTS: We show that C-terminal region of Hsf1, which consists of an activation domain AR2 and a regulatory domain CTM, mediates the Kin28-independent transcription. The AR2 domain, when fused to the DNA-binding domain of Gal4 and recruited to the GAL7 gene via the Gal4-binding sequence, is sufficient for activating GAL7 in the absence of Kin28. We have further found that AR2 has an ability to recruit TATA box-binding protein associated factors (TAFs) to the promoter. Consistently, transcription from promoters occupied naturally or artificially with TAFs is sustained in the absence of Kin28 function. CONCLUSIONS: These results show that CTM modulates activation function of AR2 in the Hsf1 molecule. We also suggest that recruitment of TAFs to a promoter is involved in the Kin28-independent transcription. PMID- 14750952 TI - Nucleocytoplasmic shuttling and phosphorylation of BMAL1 are regulated by circadian clock in cultured fibroblasts. AB - BACKGROUND: Recent discoveries of clock proteins have unveiled an important part of the mammalian circadian clock mechanism. However, the molecular clockwork that cause these fundamental feedback loops to stably oscillate with a approximately 24 h-periodicity remain unclear. RESULTS: Serum-shocked fibroblasts were used as a cellular clock model. Circadian changes in the subcellular localization and phosphorylation of BMAL1 protein in these cells were assessed by immunocytochemistry and immunoblotting. A significant time lag between Bmal1 transcription and the cytoplasmic/nuclear accumulation of BMAL1 was observed. After its nuclear accumulation, BMAL1 accumulated in the cytoplasm again, mainly by nucleoexport, before the increase of Bmal1 transcripts. Nuclear accumulation of BMAL1 matched nuclear accumulation of CLOCK and the peak of Per1 transcription. Nuclear BMAL1 was gradually phosphorylated and then dephosphorylated in a temporally regulated manner, although cytoplasmic BMAL1 was not. In serum-shocked mCry1/mCry2 (CRY)-deficient fibroblasts, which lack a functional clock, both the cytoplasmic and nuclear BMAL1 were only present as hyperphosphorylated forms and their circadian nucleocytoplasmic shuttling was absent. CONCLUSIONS: We propose that the nucleocytoplasmic shuttling and phosphorylation states of BMAL1 are regulated by circadian clock, and that this temporally regulated and time-delayed nuclear entry of BMAL1 is important in the maintenance of a stably oscillating clock. PMID- 14750953 TI - Nectin-dependent localization of synaptic scaffolding molecule (S-SCAM) at the puncta adherentia junctions formed between the mossy fibre terminals and the dendrites of pyramidal cells in the CA3 area of the mouse hippocampus. AB - BACKGROUND: Two types of intercellular junctions, synaptic junctions (SJs) and puncta adherentia junctions (PAs), are observed at the synapses between the mossy fibre terminals and the dendrites of pyramidal cells in the CA3 area of the hippocampus. SJs are associated with active zones and postsynaptic densities (PSDs) where neurotransmission occurs, whereas PAs are not associated with either of them. We have found that the nectin-afadin unit as well as the N-cadherin catenin unit localizes at the PAs and that both the units cooperatively organize the PAs. Nectins are Ca2+-independent Ig-like cell-cell adhesion molecules and afadin is a nectin- and actin filament-binding protein that connects nectins to the actin cytoskeleton. Synaptic scaffolding molecule (S-SCAM) is a neural scaffolding protein which interacts with many proteins including neuroligin, NMDA receptors, neural plakophilin-related armadillo-repeat protein/delta-catenin, a GDP/GTP exchange protein for Rap1 small G protein (PDZ-Rap-GEP), and beta catenin. S-SCAM has been suggested to be a component of PSDs, but its precise localization at the synapses remains unknown. RESULTS: S-SCAM was not concentrated at the PSDs but highly concentrated and co-localized with nectins at both the sides of the PAs formed between the mossy fibre terminals and the dendrites of pyramidal cells in the CA3 area of the adult mouse hippocampus. S SCAM co-localized with nectin-1 at the primitive synapses where the SJs and the PAs were not morphologically differentiated, and they co-localized during the maturation of the SJs and the PAs. Nectin-1 had a potency to recruit S-SCAM to the nectin-1-based cell-cell adhesion sites formed in cadherin-deficient L cells as a model system. This recruitment was dependent on the C-terminal PDZ domain binding motif of nectin-1 which is necessary for the binding of afadin, suggesting that nectins recruit S-SCAM through afadin. Consistently, S-SCAM was co-immunoprecipitated with afadin by the anti-S-SCAM antibody from the mouse brain, but S-SCAM did not directly bind afadin. CONCLUSION: These results indicate that S-SCAM localizes at the PAs in the CA3 area of the hippocampus in a nectin-dependent manner and suggest that S-SCAM serves as a scaffolding molecule at the PAs after maturation of the synapses and at the SJs during the maturation. PMID- 14750954 TI - Src-dependent phosphorylation of the EGF receptor Tyr-845 mediates Stat-p21waf1 pathway in A431 cells. AB - BACKGROUND: Cell surface receptor for the epidermal growth factor (EGFR) and cytoplasmic tyrosine kinase c-Src co-operate in several cellular functions such as proliferation and apoptosis. Our previous studies have shown that ectopic expression of the adaptor protein p52shc or p66shc, but not p46shc, and EGF stimulation lead to the activation of c-Src that is accompanied by phosphorylation of signal transducers and activators of transcription (Stat) in A431 cells. RESULTS: Here, we show that by using A431 cells as a model system, expression of p52shc, or cell stimulation with EGF or H2O2 leads to phosphorylation of EGFR on Tyr 845 that is located to the activation segment of the catalytic domain. The phosphorylation of Tyr 845 can be inhibited by PP2, but not by AG1478, and is associated with Src activation and Stat 3/5 phosphorylation, but not with MAP (mitogen-activated protein) kinase phosphorylation. Phosphorylation of Stat 3/5 in response to p52shc expression, EGF or H2O2 could also be inhibited by introduction into cells of phospho-Tyr 845 specific antibody or by expression of dominant-negative version of c-Src. Co incubation of purified c-Src and EGFR results in phosphorylation of Tyr 845 in vitro, indicating that c-Src can directly phosphorylate EGFR on Tyr 845. CONCLUSION: These results indicate that multiple signals for c-Src activation can promote Stat 3/5 phosphorylations through Src-dependent phosphorylation of EGFR on Tyr 845. PMID- 14750955 TI - Identification and characterization of a novel Dvl-binding protein that suppresses Wnt signalling pathway. AB - BACKGROUND: Dvl is a cytoplasmic protein to regulate the stability of beta catenin in the Wnt signalling pathway. However, the molecular mechanism by which Dvl regulates the Wnt signalling pathway is not fully understood. RESULTS: We identified a novel protein that binds to Dvl and named it Daple. Daple consisted of 2009 amino acids with a high frequency of leucine residues and formed a homo oligomer. The C-terminal three amino acids of Daple were necessary for binding to the region containing the PDZ domain of Dvl. Expression of Daple in mouse fibroblast L cells inhibited Wnt-3a-induced accumulation of beta-catenin. Furthermore, Daple inhibited Wnt-3a-dependent activation of T-cell factor (Tcf) transcriptional activity. Expression of Daple in the dorsal region of Xenopus embryos inhibited axis formation, which is known to be regulated by the Wnt signalling pathway. Daple also inhibited Dvl-induced secondary axis formation in Xenopus embryos. CONCLUSIONS: Daple binds to Dvl and functions as a negative regulator of the Wnt signalling pathway. PMID- 14750956 TI - Involvement of Cdc42 and Rac small G proteins in invadopodia formation of RPMI7951 cells. AB - BACKGROUND: Invadopodia are membrane protrusions into the extracellular matrix by aggressive tumour cells. These structures are associated with sites of matrix degradation and invasiveness of malignant tumour cells in an in vitro fibronectin degradation/invasion assay. The Rho family small G proteins, consisting of the Rho, Rac and Cdc42 subfamilies, are implicated in various cell functions, such as cell shape change, adhesion, and motility, through reorganization of the actin cytoskeleton. We studied the roles of the Rho family small G proteins in invadopodia formation. RESULTS: We first demonstrated that invadopodia of RPMI7951 human melanoma cells extended into the matrix substratum on a vertical view using a laser scanning confocal microscope system. We confirmed that invadopodia were rich in actin filaments (F-actin) and visualized clearly with F actin staining on a vertical view as well as on a horizontal view. We then studied the roles of Rho, Rac, and Cdc42 in invasiveness of the same cell line. In the in vitro fibronectin degradation/invasion assay, a dominant active mutant of Cdc42 enhanced dot-like degradation, whereas a dominant active mutant of Rac enhanced diffuse-type degradation. Furthermore, frabin, a GDP/GTP exchange protein for Cdc42 with F-actin-binding activity, enhanced both dot-like and diffuse-type degradation. However, a dominant active mutant of Rho did not affect the fibronectin degradation. Moreover, inhibition of phosphatidylinositol-3 kinase (PI3K) disrupted the Rac and Cdc42-dependent actin structures and blocked the fibronectin degradation. CONCLUSION: These results suggest that Cdc42 and Rac play important roles in fibronectin degradation and invasiveness in a coordinate manner through the frabin-Cdc42/Rac-PI3K signalling pathway. PMID- 14750958 TI - Control of eye orientation: where does the brain's role end and the muscle's begin? AB - Our understanding of how the brain controls eye movements has benefited enormously from the comparison of neuronal activity with eye movements and the quantification of these relationships with mathematical models. Although these early studies focused on horizontal and vertical eye movements, recent behavioural and modelling studies have illustrated the importance, but also the complexity, of extending previous conclusions to the problems of controlling eye and head orientation in three dimensions (3-D). An important facet in understanding 3-D eye orientation and movement has been the discovery of mobile, soft-tissue sheaths or 'pulleys' in the orbit which might influence the pulling direction of extraocular muscles. Appropriately placed pulleys could generate the eye-position-dependent tilt of the ocular rotation axes which are characteristic for eye movements which follow Listing's law. Based on such pulley models of the oculomotor plant it has recently been proposed that a simple two-dimensional (2 D) neural controller would be sufficient to generate correct 3-D eye orientation and movement. In contrast to this apparent simplification in oculomotor control, multiple behavioural observations suggest that the visuo-motor transformations, as well as the premotor circuitry for saccades, pursuit eye movements and the vestibulo-ocular reflexes, must include a neural controller which operates in 3 D, even when considering an eye plant with pulleys. This review summarizes the most recent work and ideas on this controversy. In addition, by proposing directly testable hypotheses, we point out that, in analogy to the previously successful steps towards elucidating the neural control of horizontal eye movements, we need a quantitative characterization first of motoneuron and next of premotor neuron properties in 3-D before we can succeed in gaining further insight into the neural control of 3-D motor behaviours. PMID- 14750959 TI - Netrin 1-mediated chemoattraction regulates the migratory pathway of LHRH neurons. AB - Luteinizing hormone-releasing hormone (LHRH) neurons migrate from the vomeronasal organ (VNO) to the forebrain in all mammals studied. In mice, the direction of LHRH neuron migration is dependent upon axons that originate in the VNO, but bypass the olfactory bulb and project caudally into the basal forebrain. Thus, factors that guide this unique subset of vomeronasal axons that comprise the caudal vomeronasal nerve (cVNN) are candidates for regulating the migration of LHRH neurons. We previously showed that deleted in colorectal cancer (DCC) is expressed by neurons that migrate out of the VNO during development [Schwarting et al. (2001) J. Neurosci., 21, 911-919]. We examined LHRH neuron migration in Dcc-/- mice and found that trajectories of the cVNN and positions of LHRH neurons are abnormal. Here we extend these studies to show that cVNN trajectories and LHRH cell migration in netrin 1 (Ntn1) mutant mice are also abnormal. Substantially reduced numbers of LHRH neurons are found in the basal forebrain and many LHRH neurons migrate into the cerebral cortex of Ntn1 knockout mice. In contrast, migration of LHRH cells is normal in Unc5h3rcm mutant mice. These results are consistent with the idea that the chemoattraction of DCC+ vomeronasal axons by a gradient of netrin 1 protein in the ventral forebrain guides the cVNN, which, in turn, determines the direction of LHRH neuron migration in the forebrain. Loss of function through a genetic deletion in either Dcc or Ntn1 results in the migration of many LHRH neurons to inappropriate destinations. PMID- 14750960 TI - Distinct functions of Rac1 and Cdc42 during axon guidance and growth cone morphogenesis in Drosophila. AB - Rho family small GTPases are thought to be key molecules in the regulation of cytoskeletal organization, especially for actin filaments. In order to examine the functions of Rac1 and Cdc42 in axon guidance at the midline of the central nervous system in Drosophila embryos, we either activated or inactivated Rac1 and Cdc42 in all postmitotic neurons. We found that the phenotypes of Cdc42 activation and Rac1 inactivation were similar to those of roundabout mutants, in that many extra axons crossed the midline. We also found that Rac1 inactivation is dominant over Roundabout receptor activation. Our observations indicate that Rac1 and Cdc42 have distinct functions in downstream signalling events triggered by Roundabout receptors. In order to further examine the functional difference between Rac1 and Cdc42 in the growth cone morphogenesis, we used primary embryonic cultures to closely observe neurite formation. We showed that activation of Rac1 and Cdc42 has distinct effects on neurite formation, particularly on growth cone morphology and the actin filaments within. Both Rac1 and Cdc42 activation induced large growth cones and long filopodia, but Cdc42 did so more efficiently than Rac1. Only Rac1 activation, however, induced thick actin bundles in the filopodia. We also found a clear difference between Rac1 and Cdc42 in terms of the response to an inhibitor of actin polymerization. Our results suggest that Cdc42 is specifically involved in the regulation of actin filaments in growth cones, whereas Rac1 is involved in additional functions. PMID- 14750961 TI - Gene expression profiling reveals multiple novel intrinsic and extrinsic factors associated with axonal regeneration failure. AB - In contrast to the regeneration-competent peripheral nervous system (PNS), lesions of nerve tracts within the central nervous system (CNS) lead to chronically impaired neuronal connections. We have analysed changes in gene expression patterns occurring as a consequence of postcommissural fornix transection at a time when spontaneous axonal growth has ceased at the lesion site. This was done in order to describe both extrinsic and intrinsic determinants of regeneration failure. Using a genomic approach we have identified a number of so far undetected factors such as bamacan and semaphorin 6B, which relate to chronic axonal growth arrest and therefore are promising candidates for lesion-induced axonal growth inhibitors. In addition, we observed that within the subiculum, where the fornix axons originate, neuronal Oct-6 was induced and NG2 was down-regulated, indicating that axotomized neurons as well as glial cells react at the level of gene expression to remote axotomy. PMID- 14750962 TI - Altered morphology of rod bipolar cell axonal terminals in the retinas of mice carrying genetic deletion of somatostatin subtype receptor 1 or 2. AB - Somatostatin (SRIF), similar to other neuropeptides, is likely to influence the morpho-functional characteristics of neurons. We studied possible morphological alterations of mouse retinal neurons following genetic deletion of SRIF subtype receptor 1 [sst1 knockout (KO)] or 2 (sst2 KO). In sst1 KO retinas, axonal terminals of rod bipolar cells (RBCs), identified with protein kinase C immunoreactivity, were 25% larger than in controls. In contrast, in sst2 KO retinas, RBC axonal terminals were significantly smaller (-14%). No major ultrastructural differences were observed between control and KO RBCs. In sst2 KO retinas, SRIF levels decreased by about 35%, while both sst1 receptor mRNA and protein increased by about 170% and 100%, respectively. This compares to previous results reporting an increase of both retinal SRIF and sst2 receptors following sst1 receptor deletion. Together, these findings suggest that, on the one hand, sst1 receptor deletion induces over-expression of sst2 receptors, and vice versa; on the other hand, that an imbalance in sst1 and sst2 receptor expression and/or changes in the levels of retinal SRIF induced by sst1 or sst2 receptor deletion are responsible for the morphological changes in RBC axonal terminals. Similar alterations of RBC terminals were observed in KO retinas at 2 weeks of age (eye opening). In addition, reverse transcription-polymerase chain reaction analysis of the expression of sst2 and sst1 receptors in developing sst1 and sst2 KO retinas, respectively, demonstrated that these receptors are up-regulated at or near eye opening. These findings suggest that the integrity of the somatostatinergic system during development is necessary for proper RBC maturation. PMID- 14750963 TI - Synthesis, localization and externalization of galectin-1 in mature dorsal root ganglion neurons and Schwann cells. AB - We recently confirmed that oxidized galectin-1 is a novel factor enhancing axonal growth in peripheral nerves after axotomy, but the process of extracellular release and oxidization of endogenous galectin-1 in the injured nervous tissue remains unknown. In the present study, we examined the distribution of galectin-1 in adult rat dorsal root ganglia (DRG) in vivo and in vitro. By RT-PCR analysis and in situ hybridization histochemistry, galectin-1 mRNA was detected in both DRG neurons and non-neuronal cells. Immunohistochemical analyses revealed that galectin-1 was distributed diffusely throughout the cytoplasm in smaller diameter neurons and Schwann cells in DRG sections. In contrast, the immunoreactivity for galectin-1 was detected in almost all DRG neurons from an early stage in culture (3 h after seeding) and was restricted to the surface and/or extracellular region of neurons and Schwann cells at later stages in culture. In a manner similar to the primary cultured cells, we also observed the surface and extracellular expression of this molecule in immortalized adult mouse Schwann cells (IMS32). Western blot analysis has revealed that both reduced and oxidized forms of galectin-1 were detected in culture media of DRG neurons and IMS32. These findings suggest that galectin-1 is externalized from DRG neurons and Schwann cells upon axonal injury. Some of the molecules in the extracellular milieu may be converted to the oxidized form, which lacks lectin activity but could act on neural tissue as a cytokine. PMID- 14750964 TI - Substance P and enkephalinergic synapses onto neurokinin-1 receptor immunoreactive neurons in the pre-Botzinger complex of rats. AB - Our previous studies have demonstrated that neurokinin-1 receptor (NK1R) immunoreactive (ir) neurons in the pre-Botzinger Complex (pre-BotC), the hypothesized kernel of respiratory rhythmogenesis, receive both glutamatergic excitatory and GABAergic or glycinergic inhibitory inputs. Neuromodulators, such as substance P (SP) and opioids, play important roles in normal respiratory activity and respiratory disorders. The identification of the relationship between neurotransmitters and NK1R-ir neurons at the cellular level is essential for understanding the synaptic interaction within the pre-BotC network. Using immunofluorescence and immunogold-silver staining, we wished to exploit SP and enkephalin (ENK) immunoreactivity and their relationships with glutamate, GABA, glycine, or NK1R in the pre-BotC in adult Sprague-Dawley rats. The pre-BotC contained a substantial amount of SP-ir and ENK-ir boutons. They were largely colocalized with glutamate and much less so with GABA. Glycine immunoreactivity was rarely found in either SP-ir or ENK-ir boutons. A number of SP-ir boutons were ENK-ir as well. Synapses were commonly found between SP-ir or ENK-ir terminals and NK1R-ir neurons in the pre-BotC. Most of them were asymmetric. Symmetric synapses made up 10% of all synapses examined between SP-ir boutons and NK1R-ir neurons, and 19% of ENK/NK1R synapses. Colocalization of SP and/or ENK with glutamate in boutons in the pre-BotC implies the combined synaptic release of excitatory amino acid and neuropeptides, which may exert combined post synaptic effects onto NK1R-ir neurons and contribute to respiratory activity. PMID- 14750965 TI - Expression of an inwardly rectifying K+ channel, Kir5.1, in specific types of fibrocytes in the cochlear lateral wall suggests its functional importance in the establishment of endocochlear potential. AB - Cochlear endolymph contains 150 mm K+ and has a highly positive potential of approximately +80 mV. The specialized ionic composition and high potential in endolymph are essential for hearing and maintained by circulation of K+ from perilymph to endolymph through the cochlear lateral wall. Various types of K+ channel such as Kir4.1 and KCNQ1/KCNE1 are expressed in stria vascularis of the lateral wall and play essential roles in K+ circulation. In this study, we examined a distribution of another K+ channel, Kir5.1, and found it specifically expressed in the spiral ligament of the cochlear lateral wall. Specific immunoreactivity for Kir5.1 was detected in type II, IV and V fibrocytes of the ligament and spiral limbus, all of which are directly involved in K+ circulation. Kir5.1 was not found in either type I or III fibrocytes. Although Kir5.1 assembles with Kir4.1 to form a functional Kir channel in renal epithelia and retinal Muller cells, double-immunolabelling revealed that they were expressed in distinct regions in the cochlea lateral wall, i.e. Kir4.1 only in stria vascularis vs. Kir5.1 in spiral ligament. During development, the expression of Kir5.1 subunits started significantly later than Kir4.1 and was correlated with the 'rapid' phase of the elevation of endocochlear potential (EP). Kir5.1 and Kir4.1 channel-subunits may therefore play distinct functional roles in K+ circulation in the cochlear lateral wall. PMID- 14750966 TI - Immunocytochemical characterization of rat brainstem neurons with vagal afferent input from the stomach challenged by acid or ammonia. AB - Exposure of the gastric mucosa to backdiffusing acid is signalled to the brainstem via vagal afferents. This study examined whether exposure of the Sprague-Dawley rat stomach to hydrochloric acid (HCl) or ammonium hydroxide (NH4OH), a noxious chemical produced by Helicobacter pylori, activates different vagal afferent pathways as reflected by different circuitries in the medullary brainstem. Two hours after intragastric treatment with HCl or NH4OH the activation of neurons in the nucleus tractus solitarii at the rostrocaudal extension of the area postrema (NTSAP) was visualized by c-Fos immunohistochemistry and their chemical coding characterized by double-labelling immunohistochemistry. Exposure of the rat gastric mucosa to HCl (0.15-0.5 M) or NH4OH (0.1-0.3 M) led to a concentration-dependent expression of c-Fos in the NTSAP. The number and distribution of NTSAP neurons activated by 0.35 M HCl and 0.3 M NH4OH were similar; the highest number of activated neurons occurring in the medial part of the NTSAP. Some 60% of the NTSAP neurons activated by intragastric HCl and NH4OH stained for the high affinity glutamate transporter EAAC1, while some 30% contained calbindin or neuropeptide Y. Glutamate receptors of the N-methyl-D-aspartate type were found on approximately 50% of the c-Fos positive cells in the NTSAP, whereas tachykinin NK1, NK2 and NK3 receptors were present on 5-10% of the activated neurons. The similar number and distribution of c-Fos-expressing neurons within the NTSAP and their identical chemical coding indicate that exposure of the rat stomach to backdiffusing concentrations of HCl and NH4OH activates the same vagal afferent-NTSAP pathway. PMID- 14750967 TI - Tissue plasminogen activator in primary afferents induces dorsal horn excitability and pain response after peripheral nerve injury. AB - The extracellular protease cascade of plasminogen activators and plasminogen are known to regulate neuronal plasticity and extracellular matrix modification, and to be important factors involved in producing long-term potentiation in the CNS. The purpose of this study is to examine the expression of plasminogen activators in primary afferents and its role in nociceptive pathways after peripheral nerve injury. We found the induction of mRNAs for tissue type plasminogen activator (tPA) and urokinase plasminogen activator (uPA) in the rat dorsal root ganglia following sciatic nerve transection. Immunoreactivity for tPA was increased in laminae I and II of the dorsal horn and, importantly, the increase in proteolytic activity mediated by tPA was observed in the same area. As neither immunoreactivity for uPA nor uPA-mediated proteolysis was observed, we further examined the effects of tPA on dorsal horn excitability and neuropathic pain behaviour. Intrathecal injection of a specific inhibitor of tPA decreased electrical stimulation-induced Fos expression in dorsal horn neurons following axotomy, and also prevented the development of thermal hyperalgesia following partial sciatic nerve ligation. These findings suggest that the increased tPA in the dorsal horn due to mRNA expression in the dorsal root ganglia increases the dorsal horn excitability and has an important role in pain behaviour after peripheral nerve injury. The tPA-mediated hypersensitivity in dorsal horn neurons may be a novel molecular mechanism of neuropathic pain. PMID- 14750968 TI - Pre- and postsynaptic contributions of voltage-dependent Ca2+ channels to nociceptive transmission in rat spinal lamina I neurons. AB - Activation of voltage-dependent Ca2+ channels (VDCCs) is critical for neurotransmitter release, neuronal excitability and postsynaptic Ca2+ signalling. Antagonists of VDCCs can be antinociceptive in different animal pain models. Neurons in lamina I of the spinal dorsal horn play a pivotal role in the processing of pain-related information, but the role of VDCCs to the activity dependent Ca2+ increase in lamina I neurons and to the synaptic transmission between nociceptive afferents and second order neurons in lamina I is not known. This has now been investigated in a lumbar spinal cord slice preparation from young Sprague-Dawley rats. Microfluorometric Ca2+ measurements with fura-2 have been used to analyse the Ca2+ increase in lamina I neurons after depolarization of the cells, resulting in a distinct and transient increase of the cytosolic Ca2+ concentration. This Ca2+ peak was reduced by the T-type channel blocker, Ni2+, by the L-type channel blockers, nifedipine and verapamil, and by the N-type channel blocker, omega-conotoxin GVIA. The P/Q-type channel antagonist, omega agatoxin TK, had no effect on postsynaptic [Ca2+]i. The NMDA receptor channel blocker D-AP5 reduced the Ca2+ peak, whereas the AMPA receptor channel blocker CNQX had no effect. Postsynaptic currents, monosynaptically evoked by electrical stimulation of the attached dorsal roots with C-fibre and Adelta-fibre intensity, respectively, were reduced by N-type channel blocker omega-conotoxin GVIA and to a much lesser extent, by P/Q-type channel antagonist omega-agatoxin TK, and the L type channel blockers verapamil, respectively. No difference was found between unidentified neurons and neurons projecting to the periaqueductal grey matter. This is the first quantitative description of the relative contribution of voltage-dependent Ca2+ channels to the synaptic transmission in lamina I of the spinal dorsal horn, which is essential in the processing of pain-related information in the central nervous system. PMID- 14750969 TI - Group I metabotropic glutamate receptors regulate the frequency-response function of hippocampal CA1 synapses for the induction of LTP and LTD. AB - Synaptically released glutamate binds to ionotropic or metabotropic glutamate receptors. Metabotropic glutamate receptors (mGluRs) are G-protein-coupled receptors and can be divided into three subclasses (Group I-III) depending on their pharmacology and coupling to signal transduction cascades. Group I mGluRs are coupled to phospholipase C and are implicated in several important physiological processes, including activity-dependent synaptic plasticity, but their exact role in synaptic plasticity remains unclear. Synaptic plasticity can manifest itself as an increase or decrease of synaptic efficacy, referred to as long-term potentiation (LTP) and long-term depression (LTD). The likelihood, degree and direction of the change in synaptic efficacy depends on the history of the synapse and is referred to as 'metaplasticity'. We provide direct experimental evidence for an involvement of group I mGluRs in metaplasticity in CA1 hippocampal synapses. Bath application of a low concentration of the specific group I agonist 3,5-dihydroxyphenylglycine (DHPG), which does not affect basal synaptic transmission, resulted in a leftward shift of the frequency-response function for the induction of LTD and LTP in naive synapses. DHPG resulted in the induction of LTP at frequencies which induced LTD in control slices. These alterations in the induction of LTD and LTP resemble the metaplastic changes observed in previously depressed synapses. In addition, in the presence of DHPG additional potentiation could be induced after LTP had apparently been saturated. These findings provide strong evidence for an involvement of group I mGluRs in the regulation of metaplasticity in the CA1 field of the hippocampus. PMID- 14750970 TI - Diversity beyond variance: modulation of firing rates and network coherence by GABAergic subpopulations. AB - Computational modelling studies have revealed that heterogeneity in interneuronal networks can powerfully modulate firing rates, responses to excitatory inputs, theta-gamma oscillations and network synchrony. In these previous studies, heterogeneity was represented by the degree of variance in various interneuronal synaptic and cellular parameters. However, a major characteristic of gamma-amino butyric-acid-ergic (GABAergic) synaptic and interneuronal populations is the presence of distinct subgroups, which variance-based approaches cannot fully address. Here we apply an information theory-based measure of diversity, the Shannon-Wiener diversity index (equivalent to entropy), which takes into account both the number and relative abundance of categories within a population. Computational modelling data and experimental dynamic clamp results show that increasing the diversity index in the somatically injected inhibitory post synaptic currents (IPSC) peak conductances modulates the firing rates of CA1 pyramidal cells in a predictable manner that depends on both the mean and variance of the IPSC conductance. Furthermore, increases in the diversity index in interneuronal populations strongly decreased network coherence, even when population variance, the previously applied measure of heterogeneity, remained unchanged. These modelling and experimental results reveal a new approach to the study of interneuronal heterogeneity, and demonstrate the modulation of principal cell firing rates and interneuronal network coherence by parameter clustering at both the synaptic and cellular levels. PMID- 14750971 TI - Suppressed proliferation and apoptotic changes in the rat dentate gyrus after acute and chronic stress are reversible. AB - Acute stress suppresses new cell birth in the hippocampus in several species. Relatively little is known, however, on how chronic stress affects the turnover, i.e. proliferation and apoptosis, of the rat dentate gyrus (DG) cells, and whether the stress effects are lasting. We investigated how 3 weeks of chronic unpredictable stress would influence the structural dynamic plasticity of the rat DG, and studied newborn cell proliferation, survival, apoptosis, volume and cell number in 10-week-old animals. To study lasting effects, another group of animals was allowed to recover for 3 weeks. Based on two independent parameters, bromodeoxyuridine (BrdU) and Ki-67 immunocytochemistry, our results show that both chronic and acute stress decrease new cell proliferation rate. The reduced proliferation after acute stress normalized within 24 h. Interestingly, chronically stressed animals showed recovery after 3 weeks, albeit with still fewer proliferating cells than controls. Apoptosis, by contrast, increased after acute but decreased after chronic stress. These results demonstrate that, although chronic stress suppresses proliferation and apoptosis, 3 weeks of recovery again normalized most of these alterations. This may have important implications for our understanding of the reversibility of stress-related hippocampal volume changes, such as occur, for example, in depression. PMID- 14750972 TI - The opposite effects of stress on dendritic spines in male vs. female rats are NMDA receptor-dependent. AB - Dendritic spines in the hippocampus are sources of synaptic contact that may be involved in processes of learning and memory [Moser (1999) Cell. Mol. Life Sci., 55, 593-600]. These structures are sensitive to sex differences as females in proestrus possess a greater density than males and females in other stages of the estrous cycle [Woolley et al. (1990) J. Neurosci., 10, 4035-4039]. Moreover, exposure to an acute stressful event increases spine density in the male hippocampus but decreases spine density in the female hippocampus [Shors et al. (2001) J. Neurosci., 21, 6292-6297]. Here we demonstrate that antagonism of N methyl-d-aspartate (NMDA) receptors prevents the increase in spine density as females transition from diestrus 2 to proestrus, when estrogen levels are rising. Antagonism of NMDA receptors during exposure to the stressful event also prevented the changes in spine density in males and females, despite differences in the direction of these effects. Thus, the stress-induced increase in spine density was prevented in the male hippocampus as was the stress-induced decrease in spine density in the female hippocampus. NMDA receptor antagonism during exposure to the stressful event did not alter corticosterone levels or the corticosterone response to stress. These data suggest that both increases and decreases in spine density can be dependent on NMDA receptor activation. PMID- 14750973 TI - Behavioural adaptations to addictive drugs in mice lacking the NMDA receptor epsilon1 subunit. AB - N-methyl-D-aspartate (NMDA) receptors, a subtype of glutamate receptors (GluRs) formed by assembly of the GluRzeta subunit (called NR1 in rats) with any one of four GluRepsilon subunits (GluRepsilon1-4; NR2A-D), play an important role in excitatory neurotransmission, synaptic plasticity and brain development. Recent pharmacological studies have also indicated a role for NMDA receptors in drug addiction. In the present study, we investigated the behavioural adaptations to addictive drugs such as phencyclidine (PCP), methamphetamine (MAP) and morphine (MOR) in mice lacking the GluRepsilon1 subunit of the NMDA receptor. GluRepsilon1 mutant mice exhibited a malfunction of NMDA receptors, as evidenced by the reduction of [3H]MK-801 binding in an autoradiographic receptor binding assay. GluRepsilon1 mutant mice showed an attenuation of acute PCP- and MAP-induced hyperlocomotion. The development of sensitization by repeated treatment with PCP and MAP at a low, but not high, dose was also suppressed. The development of MOR induced analgesic tolerance and naloxone-precipitated MOR withdrawal symptoms were attenuated in GluRepsilon1 mutant mice. In the place conditioning test, PCP induced place aversion in naive mice and place preference in PCP-pretreated mice, as well as MOR-induced place preference, were diminished whereas MAP-induced place preference was not affected in GluRepsilon1 mutant mice. These findings provide genetic evidence that GluRepsilon1 subunit-containing NMDA receptors are involved in certain aspects of drug addiction. PMID- 14750974 TI - Environmental enrichment affects striatal graft morphology and functional recovery. AB - Environmental conditions and behavioural experience can affect neuronal function and morphology. It is less well known whether such factors also influence the growth, integration and functional recovery provided by neural grafts placed within the damaged brain. Here we report on the effects of differential housing conditions on striatal graft morphology and functional recovery after striatal lesions. Rats were pretrained on a skilled bilateral forelimb task, the staircase test, and lesioned unilaterally in the lateral dorsal striatum with quinolinic acid. One group of lesioned animals was given suspension grafts of E15 whole ganglionic eminence implanted into the lesioned striatum. Following transplantation, the animals were housed either in standard cages (four per cage) or in enriched environment housing conditions (10 per cage) with tunnels, ladders and increased living space available for exploration, social interaction and play. The differentially housed animals were retested on the skilled staircase test at two separate time points. Repeated testing, environmental enrichment and transplantation positively influenced behavioural recovery. Partial recovery was observed bilaterally amongst the grafted animals in both housing conditions. Nevertheless, the grafted animals housed in the enriched environment performed significantly better in the final test compared with all of the other experimental groups. The grafts survived equally well under both housing conditions but the grafts of animals housed in the enriched environment contained larger projection neurons and were somewhat better reinnervated by dopaminergic afferents. An increased level of striatal brain-derived neurotrophic factor was observed in the control animals housed under the enriched compared with the standard conditions. The results indicate that an enriched environment can affect both graft function and graft morphology through as yet unknown mechanisms. PMID- 14750975 TI - First- and second-order stimulus length selectivity in New World monkey striate cortex. AB - Motion is a powerful cue for figure-ground segregation, allowing the recognition of shapes even if the luminance and texture characteristics of the stimulus and background are matched. In order to investigate the neural processes underlying early stages of the cue-invariant processing of form, we compared the responses of neurons in the striate cortex (V1) of anaesthetized marmosets to two types of moving stimuli: bars defined by differences in luminance, and bars defined solely by the coherent motion of random patterns that matched the texture and temporal modulation of the background. A population of form-cue-invariant (FCI) neurons was identified, which demonstrated similar tuning to the length of contours defined by first- and second-order cues. FCI neurons were relatively common in the supragranular layers (where they corresponded to 28% of the recorded units), but were absent from layer 4. Most had complex receptive fields, which were significantly larger than those of other V1 neurons. The majority of FCI neurons demonstrated end-inhibition in response to long first- and second-order bars, and were strongly direction selective. Thus, even at the level of V1 there are cells whose variations in response level appear to be determined by the shape and motion of the entire second-order object, rather than by its parts (i.e. the individual textural components). These results are compatible with the existence of an output channel from V1 to the ventral stream of extrastriate areas, which already encodes the basic building blocks of the image in an invariant manner. PMID- 14750976 TI - Lesions of dorsolateral striatum preserve outcome expectancy but disrupt habit formation in instrumental learning. AB - Habits are controlled by antecedent stimuli rather than by goal expectancy. Interval schedules of feedback have been shown to generate habits, as revealed by the insensitivity of behaviour acquired under this schedule to outcome devaluation treatments. Two experiments were conducted to assess the role of the dorsolateral striatum in habit learning. In Experiment 1, sham operated controls and rats with dorsolateral striatum lesions were trained to press a lever for sucrose under interval schedules. After training, the sucrose was devalued by inducing taste aversion to it using lithium chloride, whereas saline injections were given to the controls. Only rats given the devaluation treatment reduced their consumption of sucrose and this reduction was similar in both the sham and the lesioned groups. All rats were then returned to the instrumental chamber for an extinction test, in which the lever was extended but no sucrose was delivered. In contrast to sham operated controls, rats with dorsolateral striatum lesions refrained from pressing the lever if the outcome was devalued. To assess the specificity of the role of dorsolateral striatum in this effect a second experiment was conducted in which a group with lesions of dorsomedial striatum was added. In relation now to both the sham and the dorsomedial lesioned groups, only rats with lesions of dorsolateral striatum significantly reduced responding after outcome devaluation. In conclusion, this study provides direct evidence that the dorsolateral striatum is necessary for habit formation. Furthermore, it suggests that, when the habit system is disrupted, control over instrumental performance reverts to the system controlling the performance of goal-directed instrumental actions. PMID- 14750977 TI - Execution and observation of bringing a fruit to the mouth affect syllable pronunciation. AB - Kinematic analysis of lip and voice spectrum analysis were used to assess the influence of both execution and observation of arm-mouth-related actions on speech production. In experiments 1 and 2 participants brought either a cherry or an apple to their mouth and either pronounced the syllable BA (experiment 1) or emitted a nonspeech-related vocalization (experiment 2). In the other three experiments participants observed arm actions performed by the experimenter and pronounced the syllable BA. In experiment 3, they observed the action of bringing the cherry or apple to the mouth. In experiments 4 and 5, they observed a pantomime of the same action performed by the experimenter with his own arm (experiment 4) or with a nonbiological arm (experiment 5). The results showed that the formant 2 of the vowel 'a' increased when participants executed the bringing-to-the-mouth act with the apple or observed its execution or pantomime with the experimenter's arm (experiments 1, 3 and 4). In contrast, no modification in the vowel formants was found during a nonspeech-related vocalization (experiment 2) and during observation of an action with a nonbiological arm (experiment 5). Finally, the opening of the lips was larger when the participant brought the apple rather than the cherry to the mouth and pronounced BA (experiment 1). Taken together, the results of the present study suggest that the execution and observation of the bringing-to-the-mouth action activate a mouth articulation posture (probably due to the act of food manipulation with the mouth) which selectively influences speech production. They support the idea that the system involved in speech production shares and may derive from the neural substrate which is involved in the control of arm-mouth interactions and, in general, of arm actions. PMID- 14750978 TI - Stimulus-induced change in long-range temporal correlations and scaling behaviour of sensorimotor oscillations. AB - The human brain spontaneously generates large-scale network oscillations at around 10 and 20 Hz. The amplitude envelope of these oscillations fluctuates intermittently and was recently reported to exhibit power-law decay of the autocorrelation for hundreds of seconds. This indicates that the underlying networks are in a dynamic state resembling the self-organized critical state known to exist in many complex systems. Based on the mechanism of how correlations emerge in these systems, we hypothesized that the physiological basis of long-range power-law correlations is the buildup of a memory of past activity by a continuous modification of the network's functional connectivity by the ongoing oscillations. In this framework, exogenous perturbations of ongoing oscillations would degrade or abolish this dynamic network memory. We investigated the sensitivity of the temporal correlations in sensorimotor 10- and 20-Hz oscillations to median nerve stimulation that is known to have immediate effects on ongoing oscillations. Our results show that the amplitude fluctuations of these oscillations were effectively modulated by the somatosensory stimuli but still exhibited long-range temporal correlations and power-law scaling behaviour. The magnitude of the temporal correlations was, however, attenuated and the power law exponents were decreased. This implies that the stimuli indeed degraded the network's memory of its past. PMID- 14750979 TI - Molecular cloning and expression regulation of PRG-3, a new member of the plasticity-related gene family. AB - Phospholipid-mediated signalling on neurons provokes diverse responses such as neurogenesis, pattern formation and neurite remodelling. We have recently uncovered a novel set of molecules in the mammalian brain, named plasticity related genes (PRGs), which mediate lipid phosphate phosphatase activity and provide evidence for their involvement in mechanisms of neuronal plasticity. Here, we report on a new member of the vertebrate-specific PRG family, which we have named plasticity-related gene-3 (PRG-3). PRG-3 is heavily expressed in the brain and shows a specific expression pattern during brain development where PRG 3 expression is found predominantly in neuronal cell layers and is already expressed at embryonic day 16. In the mature brain, strongest PRG-3 expression occurs in the hippocampus and cerebellum. Overexcitation of neurons induced by kainic acid leads to a transient down-regulation of PRG-3. Furthermore, PRG-3 is expressed on neurite extensions and promotes neurite growth and a spreading-like cell body in neuronal cells and COS-7 cells. In contrast to previously described members of the PRG family, PRG-3 does not perform its function through enzymatic phospholipid degradation. In summary, our findings feature a new member of the PRG family which shows dynamic expression regulation during brain development and neuronal excitation. PMID- 14750980 TI - Effect of gene transfer of GLT-1, a glutamate transporter, into the locus coeruleus by recombinant adenoviruses on morphine physical dependence in rats. AB - There is a body of evidence implying the involvement of the central glutamatergic system in morphine dependence. We previously reported changes in the mRNA expression of a glial glutamate transporter GLT-1 in some brain regions of morphine-dependent and naloxone-precipitated withdrawal rats, and inhibition of morphine physical dependence by a glutamate transporter activator in mice. These findings support the possibility that glutamate transporters such as GLT-1 are involved in morphine dependence. In this study, we examined the effects of gene transfer of GLT-1 into the locus coeruleus (LC) by recombinant adenoviruses on morphine physical dependence in rats. We constructed recombinant adenoviruses that successfully delivered the GLT-1 gene in vitro and in vivo. Local overexpression of GLT-1 within the bilateral LC by the recombinant adenoviruses before implantation of the morphine pellet significantly inhibited various somatic signs of naloxone-precipitated morphine withdrawal, compared with the control. These results suggest that GLT-1 within the LC plays an inhibitory role in morphine physical dependence. PMID- 14750981 TI - Ventricular beat detection in single channel electrocardiograms. AB - BACKGROUND: Detection of QRS complexes and other types of ventricular beats is a basic component of ECG analysis. Many algorithms have been proposed and used because of the waves' shape diversity. Detection in a single channel ECG is important for several applications, such as in defibrillators and specialized monitors. METHODS: The developed heuristic algorithm for ventricular beat detection includes two main criteria. The first of them is based on steep edges and sharp peaks evaluation and classifies normal QRS complexes in real time. The second criterion identifies ectopic beats by occurrence of biphasic wave. It is modified to work with a delay of one RR interval in case of long RR intervals. Other algorithm branches classify already detected QRS complexes as ectopic beats if a set of wave parameters is encountered or the ratio of latest two RR intervals RRi-1/RRi is less than 1:2.5. RESULTS: The algorithm was tested with the AHA and MIT-BIH databases. A sensitivity of 99.04% and a specificity of 99.62% were obtained in detection of 542014 beats. CONCLUSION: The algorithm copes successfully with different complicated cases of single channel ventricular beat detection. It is aimed to simulate to some extent the experience of the cardiologist, rather than to rely on mathematical approaches adopted from the theory of signal analysis. The algorithm is open to improvement, especially in the part concerning the discrimination between normal QRS complexes and ectopic beats. PMID- 14750982 TI - Primary Malignant Melanoma of the Lung: A Case Report. AB - BACKGROUND: Primary melanoma of the lung is an extremely rare pathological entity and sparsely reported in the literature. CASE PRESENTATION: A case of primary malignant melanoma of the lung in a 41-year-old female is reported. The clinical, radiological and histopathological features are discussed. The initial symptom was cough, whereas the chest radiography showed a round opacity of the right lung. The computed tomography of the chest revealed a well-demarcated mass lesion in the right upper lobe. Endobronchial mass causing obstruction of the upper lobar bronchus was the bronchoscopic finding. Patient underwent pneumonectomy. A diagnosis of melanoma was confirmed postoperatively after the immunohistochemistry. Primary nature of the tumour in the lung results from the demonstration of characteristic junctional pattern of melanoma cells beneath the bronchial epithelium on histopathology, and from exclusion of other potential primary sites in the clinical, paraclinical and laboratory examination. CONCLUSIONS: Primary melanoma of the lung represents a rare pathological entity. Careful interpretation of histopathological information in correlation with all other findings from clinical and paraclinical studies can establish a diagnosis. Follow-up is necessary in order to diagnose potential dissemination or secondary sites of the disease. Due to the small number of cases reported in the literature, there is no experience on the management and the prognosis of the disease, but surgical resection remains the cornerstone of the treatment. PMID- 14750983 TI - Letter to the Editor: comments on "Compositional averaging of backscatter intensities in compounds.". PMID- 14750984 TI - Thin dielectric film thickness determination by advanced transmission electron microscopy. AB - High-resolution transmission electron microscopy (HR-TEM) has been used as the ultimate method of thickness measurement for thin films. The appearance of phase contrast interference patterns in HR-TEM images has long been confused as the appearance of a crystal lattice by nonspecialists. Relatively easy to interpret crystal lattice images are now directly observed with the introduction of annular dark-field detectors for scanning TEM (STEM). With the recent development of reliable lattice image processing software that creates crystal structure images from phase contrast data, HR-TEM can also provide crystal lattice images. The resolution of both methods has been steadily improved reaching now into the sub Angstrom region. Improvements in electron lens and image analysis software are increasing the spatial resolution of both methods. Optimum resolution for STEM requires that the probe beam be highly localized. In STEM, beam localization is enhanced by selection of the correct aperture. When STEM measurement is done using a highly localized probe beam, HR-TEM and STEM measurement of the thickness of silicon oxynitride films agree within experimental error. In this article, the optimum conditions for HR-TEM and STEM measurement are discussed along with a method for repeatable film thickness determination. The impact of sample thickness is also discussed. The key result in this article is the proposal of a reproducible method for film thickness determination. PMID- 14750985 TI - Using fluctuation microscopy to characterize structural order in metallic glasses. AB - We have used fluctuation microscopy to reveal the presence of structural order on length scales of 1-2 nm in metallic glasses. We compare results of fluctuation microscopy measurements with high resolution transmission electron microscopy and electron diffraction observations on a series of metallic glass samples with differing degrees of structural order. The agreement between the fluctuation microscopy results and those of the other techniques is good. In particular, we show that the technique used to make thin specimens for electron microscopy affects the structure of the metallic glass, with ion thinning inducing more structural order than electro-polishing. We also show that relatively minor changes in the composition of the alloy can have a significant effect on the medium-range order; this increased order is correlated with changes in mechanical behavior. PMID- 14750986 TI - Selected-area growth of carbon nanotubes by the combination of focused ion beam and chemical vapor deposition techniques. AB - In this article, we report a technique for growing carbon nanotubes in a more controllable fashion, which enables us to synthesize nanotubes directly in various forms of designed patterns. This nanofabrication process is based on a combination of focused ion beam (FIB) and chemical vapor deposition (CVD) techniques. In this process, arrays of conductive patterns were first deposited on silicon substrates by directing a gaseous compound (C(9)H(16)Pt) via the capillary needle-sized nozzles within a FIB system. The substrates were then coated with catalyst and further modified by the FIB to localize the position of the catalyst. Finally, the growth of carbon nanotubes on the designed substrates was carried out by CVD of hydrocarbon gases. This fabrication technique has the advantage of positioning carbon nanotubes in selected locations. This may open up opportunities for the direct synthesis of carbon nanotubes onto almost any substrate material, thus allowing fabrication of carbon nanotube-based devices. PMID- 14750987 TI - The use of high pressure freezing and freeze substitution to study host-pathogen interactions in fungal diseases of plants. AB - This article reports on the use of high pressure freezing followed by freeze substitution (HPF/FS) to study ultrastructural details of host-pathogen interactions in fungal diseases of plants. The specific host-pathogen systems discussed here include a powdery mildew infection of poinsettia and rust infections of daylily and Indian strawberry. The three pathogens considered here all attack the leaves of their hosts and produce specialized hyphal branches known as haustoria that invade individual host cells without killing them. We found that HPF/FS provided excellent preservation of both haustoria and host cells for all three host-pathogen systems. Preservation of fungal and host cell membranes was particularly good and greatly facilitated the detailed study of host-pathogen interfaces. In some instances, HPF/FS provided information that was not available in samples prepared for study using conventional chemical fixation. On the other hand, we did encounter various problems associated with the use of HPF/FS. Examples included freeze damage of samples, inconsistency of fixation in different samples, separation of plant cell cytoplasm from cell walls, breakage of cell walls and membranes, and splitting of thin sections. However, we believe that the outstanding preservation of ultrastructural details afforded by HPF/FS significantly outweighs these problems and we highly recommend the use of this fixation protocol for future studies of fungal host-plant interactions. PMID- 14750988 TI - Angioarchitecture of tumors induced by two different cloned cell lines established from a transplantable rat malignant fibrous histiocytoma. AB - Angiogenesis, a biologic process whereby endothelial cells divide and migrate to form new blood vessels, is a key step in tumor growth, invasion, and metastasis. In the present study, we investigated the differences in angioarchitecture between two different tumors induced by cloned cell lines (MT-8 and MT-9), derived from a transplantable rat malignant fibrous histiocytoma, by scanning electron microscopy of vascular corrosion casts. During a 3-week observation period after implantation, the growth of MT-8 tumors appeared to be faster than that of MT-9 tumors. Histologically, MT-8 tumors were of the uniformly undifferentiated sarcoma type arranged in characteristic organoid structures, and MT-9 tumors showed a storiform growth pattern. In MT-8 tumors, neovascularization occurred by sprouting at postimplantation (PI) week 1, and the newly formed capillaries gradually became more tortuous. In MT-9 tumors, at PI week 1, the corrosion casts of newly formed capillaries mainly showed a wavy course but no finger-like outgrowths of capillaries were seen. At PI weeks 2 and 3, the sprouting was seen specifically in MT-9 tumors, forming basket-like structures and glomeruloid structures of capillaries. These results indicate that angiogenesis or angioarchitecture of MT-8 tumors is different from that of MT-9 tumors, depending on the differences in their tumor histology and by the features like absence or presence of basket-like structures and glomeruloid structures of capillaries. PMID- 14750989 TI - Extending energy-filtered transmission electron microscopy (EFTEM) into three dimensions using electron tomography. AB - The length scales on which materials microstructures are being formed, grown, and even designed are becoming increasingly small and increasingly three-dimensional. For such complex structures two-dimensional transmission electron microscopy (TEM) analysis is often inadequate and occasionally misleading. One approach to this problem is the modification of electron tomography techniques, developed for structural biology, for use in materials science. Energy-Filtered (EF) TEM elemental distribution images approximate to true projections of structure, and, as such, can be used to reconstruct the three-dimensional distribution of chemical species. A sample holder has been modified to allow the high tilt (+/-60 degrees ) required for tomography and a semiautomatic acquisition script designed to manage energy-loss acquisition. Tilt series data sets have been acquired from two widely different experimental systems, Cr carbides in 316 stainless steel and magnetite nanocrystals in magnetotactic bacteria, demonstrating single- and multiple-element tomography. It is shown that both elemental maps and jump-ratio images are suitable for reconstruction, despite the effects of diffraction contrast in the former and thickness changes in the latter. It is concluded that the image contrast, signal, and signal-to-noise ratio (SNR) are key to the achievable reconstruction quality and, as such, the technique may be of limited value for high energy loss/small inelastic cross section edges. PMID- 14750990 TI - Object oriented database and electronic notebook for transmission electron microscopy. AB - As high-resolution biological transmission electron microscopy (TEM) has increased in popularity over recent years, the volume of data and number of projects underway has risen dramatically. A robust tool for effective data management is essential to efficiently process large data sets and extract maximum information from the available data. We present the Electron Microscopy Electronic Notebook (EMEN), a portable, object-oriented, web-based tool for TEM data archival and project management. EMEN has several unique features. First, the database is logically organized and annotated so multiple collaborators at different geographical locations can easily access and interpret the data without assistance. Second, the database was designed to provide flexibility to the user, so it can be used much as a lab notebook would be, while maintaining a structure suitable for data mining and direct interaction with data-processing software. Finally, as an object-oriented database, the database structure is dynamic and can be easily extended to incorporate information not defined in the original database specification. PMID- 14750991 TI - Electron beam coater for reduction of charging in ice-embedded biological specimens using Ti(88)Si(12) alloy. AB - Biological macromolecules embedded in vitreous ice are known to suffer from charging while being imaged in an electron transmission cryomicroscope. We developed an electron beam coater that deposits conductive films onto the surface of frozen-hydrated specimens. The conductive films help to dissipate charge during electron irradiation of poorly conductive ice-embedded biological samples. We observed significant reduction in charging of ice-embedded catalase crystals suspended over holes in a holey carbon film after coating them with a 30-A-thick layer of an amorphous alloy, Ti(88)Si(12). Images of the crystals after coating showed diffraction spots of up to 3 A resolution. PMID- 14750992 TI - Analysis of orientations of collagen fibers by novel fiber-tracking software. AB - Recent evidence supports the notion that biological functions of extracellular matrix (ECM) are highly correlated to not only its composition but also its structure. This article integrates confocal microscopy imaging and image processing techniques to analyze the microstructural properties of ECM. This report describes a two- and three-dimensional fiber middle-line tracing algorithm that may be used to quantify collagen fibril organization. We utilized computer simulation and statistical analysis to validate the developed algorithm. These algorithms were applied to confocal images of collagen gels made with reconstituted bovine collagen type I, to demonstrate the computation of orientations of individual fibers. PMID- 14751002 TI - Methamphetamine dependence increases risk of neuropsychological impairment in HIV infected persons. AB - Both HIV infection and methamphetamine dependence can be associated with brain dysfunction. Little is known, however, about the cognitive effects of concurrent HIV infection and methamphetamine dependence. The present study included 200 participants in 4 groups: HIV infected/methamphetamine dependent (HIV+/METH+), HIV negative/methamphetamine dependent (HIV-/METH+), HIV infected/methamphetamine nondependent (HIV+/METH-), and HIV negative/methamphetamine nondependent (HIV /METH-). Study groups were comparable for age, education, and ethnicity, although the HIV-/METH- group had significantly more females. A comprehensive, demographically corrected neuropsychological battery was administered yielding a global performance score and scores for seven neurobehavioral domains. Rates of neuropsychological impairment were determined by cutoff scores derived from performances of a separate control group and validated with larger samples of HIV+ and HIV- participants from an independent cohort. Rates of global neuropsychological impairment were higher in the HIV+/METH+ (58%), HIV-/METH+ (40%) and HIV+/METH- (38%) groups compared to the HIV-/METH- (18%) group. Nonparametric analyses revealed a significant monotonic trend for global cognitive status across groups, with least impairment in the control group and highest prevalence of impairment in the group with concurrent HIV infection and methamphetamine dependence. The results indicate that HIV infection and methamphetamine dependence are each associated with neuropsychological deficits, and suggest that these factors in combination are associated with additive deleterious cognitive effects. This additivity may reflect common pathways to neural injury involving both cytotoxic and apoptotic mechanisms. PMID- 14751003 TI - Dopamine and semantic activation: an investigation of masked direct and indirect priming. AB - To investigate the effects of dopamine on the dynamics of semantic activation, 39 healthy volunteers were randomly assigned to ingest either a placebo (n = 24) or a levodopa (n = 16) capsule. Participants then performed a lexical decision task that implemented a masked priming paradigm. Direct and indirect semantic priming was measured across stimulus onset asynchronies (SOAs) of 250, 500 and 1200 ms. The results revealed significant direct and indirect semantic priming effects for the placebo group at SOAs of 250 ms and 500 ms, but no significant direct or indirect priming effects at the 1200 ms SOA. In contrast, the levodopa group showed significant direct and indirect semantic priming effects at the 250 ms SOA, while no significant direct or indirect priming effects were evident at the SOAs of 500 ms or 1200 ms. These results suggest that dopamine has a role in modulating both automatic and attentional aspects of semantic activation according to a specific time course. The implications of these results for current theories of dopaminergic modulation of semantic activation are discussed. PMID- 14751004 TI - Mapping the neural systems that mediate the Paced Auditory Serial Addition Task (PASAT). AB - The paced auditory serial addition task (PASAT), in which subjects hear a number string and add the two most-recently heard numbers, is a neuropsychological test sensitive to cerebral dysfunction. We mapped the brain regions activated by the PASAT using positron emission tomography (PET) and 15O-water to measure cerebral blood flow. We parsed the PASAT by mapping sites activated by immediate repetition of numbers and by repetition of the prior number after the presentation of the next number in the series. The PASAT activated dispersed non contiguous foci in the superior temporal gyri, bifrontal and biparietal sites, the anterior cingulate and bilateral cerebellar sites. These sites are consistent with the elements of the task that include auditory perception and processing, speech production, working memory, and attention. Sites mediating addition were not identified. The extent of the sites activated during the performance of the PASAT accounts for the sensitivity of this test and justifies its use in a variety of seemingly disparate conditions. PMID- 14751005 TI - Working memory and processing speed deficits in systemic lupus erythematosus as measured by the paced auditory serial addition test. AB - As many as 66% of systemic lupus erythematosus (SLE) patients have been reported to have cognitive deficits. These deficits are often associated with information processing speed and working memory. Similarly, processing speed and working memory impairments are the hallmark of cognitive dysfunction in multiple sclerosis (MS). The Paced Auditory Serial Addition Test (PASAT) places high demands on processing speed and working memory. Fisk and Archibald, however, demonstrated that the total score of the PASAT does not accurately reflect impairments in these cognitive processes. They found that MS patients used a chunking strategy to obtain correct responses and reduce the cognitive demands of the task. In the present study, PASAT performance was examined for 45 SLE patients and 27 controls using alternative scoring procedures. Although the total number of correct responses did not differ between SLE and controls at the 2.4 or 2.0 s presentation rates, SLE patients had fewer dyads (correct consecutive responses) than controls at the faster rate, and more chunking responses than controls at both rates. Disease activity, disease duration, depression, fatigue, and corticosteroids could not account for these differences. The findings suggest that SLE patients, like MS patients, chunk responses more often than controls, and that this scoring procedure may better reflect the working memory and processing speed deficits present in SLE. PMID- 14751006 TI - Confrontation naming in Chinese patients with left, right or bilateral brain damage. AB - Confrontation naming of 52 unilateral or bilateral brain-damaged Chinese patients were examined with a modified version of the Boston Naming Test (BNT). Chinese patients with left or right hemisphere lesions, contrary to studies on English speakers, demonstrated similar levels of naming impairments, supporting the notion that English and Chinese are mediated by different neuroprocessing systems. In addition, the psychometric properties of the BNT on Chinese population were examined. While the test demonstrated satisfactory internal consistency and discriminant validity, level of education was found to be a significant factor affecting participants' performance. A cut-off score of 24 in spontaneous naming yielded a sensitivity of 73.1% and specificity of 75.3% in differentiating normal from brain-damaged participants, suggesting that the modified BNT is applicable to the Chinese population. PMID- 14751007 TI - Inhibition of return in obsessive-compulsive disorder. AB - Obsessive-compulsive disorder (OCD) is characterized by repetitive obsessions and/or compulsions that interfere with daily functioning. Neuropsychological studies have suggested that such perseverative behaviors may be due to underlying attentional deficits. Inhibition of return (IOR) is an adaptive mechanism that is thought to assist visual search by biasing attention after a critical, short interval to novel, previously unattended areas. Therefore, this study aimed to examine whether deficient IOR mechanisms could underlie some of the attentional, and perhaps behavioral, problems, reported in OCD patients. Using a computerized IOR paradigm, participants were required to respond to a target that appeared at either the same or different location to a precue that was presented either 100 ms or 700 ms earlier. Results indicate that patients had a reduced IOR for targets presented in the left visual field, suggesting lateralized anomalies in shifting attention. Results are consistent with lateralization anomalies previously reported in OCD. PMID- 14751008 TI - Effects of hypoxia on the brain: neuroimaging and neuropsychological findings following carbon monoxide poisoning and obstructive sleep apnea. AB - Hypoxia damages multiple organ systems especially those with high oxygen utilization such as the central nervous system. The purpose of this study was to compare the neuropathological and neuropsychological effects of hypoxia in patients with either carbon monoxide poisoning or obstructive sleep apnea. Neuroimaging revealed evidence of hippocampal atrophy in both groups although a linear relationship between hippocampal volume and memory performance was found only for selected tests and only in the sleep apnea group. There were significant correlations between hippocampal volume and performance on measures related to nonverbal/information processing. Generalized brain atrophy, as measured by the ventricle-to-brain ratio, was more common in the carbon monoxide poisoning group compared to the obstructive sleep apnea group. Performance on tests of executive function improved following treatment with nasal continuous positive airway pressure treatment in the obstructive sleep apnea group but there was no associated improvement in general intellectual function. We found that hypoxia due to obstructive sleep apnea and CO poisoning resulted in neuropathological changes and neuropsychological impairments. The observed group differences provide insight into the relationship between etiology of injury, neuropathological changes, and clinical presentation. PMID- 14751009 TI - A normative study of the CERAD neuropsychological assessment battery in the Korean elderly. AB - This study aimed to explore the effects of age, education and gender on the performance of eight tests in the Korean version of the CERAD neuropsychological assessment battery and to provide normative information on the tests in the Korean elderly. The battery was administered to 618 healthy volunteers aged from 60 to 90. People with serious neurological, medical and psychiatric disorders, including dementia, were excluded. Multiple linear regression analyses were performed to assess the relative contribution of the demographic factors on the score of each cognitive test. Age, education, and gender were found to have significant effects on the performance of many tests in the battery. Based on these results, 4 overlapping age normative tables (60 to 74, 65 to 79, 70 to 84, and 75 to 90 years of age) with 3 educational strata (0 to 3 years, 4 to 6 years, and 7 years and more) for both genders are presented. The normative information will be useful for a clinical interpretation of the CERAD neuropsychological battery in Korean elderly as well as for comparing the performance of the battery across countries. PMID- 14751010 TI - How well does IQ predict neuropsychological test performance in normal adults? AB - The strength and nature of the association between IQ and performance on other cognitive tests has both practical and conceptual significance for clinical neuropsychology. In this study, 28 measures derived from 16 cognitive tests were analyzed as a function of IQ in 221 adults. Participants were grouped by their IQ scores as having below average (BA), average (A), or above average (AA) intelligence. Planned comparisons revealed that A adults performed significantly better than BA adults on 25 of the 28 cognitive measures, and that AA adults performed significantly better than A adults on 19 of 28 measures. Effect sizes averaged.74 for BA-A comparisons and.41 for A-AA comparisons. Linear, quadratic, and cubic functions described the relationships between IQ and cognitive test performance equally well for most individual test measures and for a composite index of test performance, whereas quadratic and cubic functions explained the proportion of abnormal performances better than a linear function. These findings confirm that IQ predicts concurrent neuropsychological performance across the entire spectrum of intelligence, but more so among persons of average IQ or less than among those with above average IQ. PMID- 14751011 TI - Semantic interference deficits and the detection of mild Alzheimer's disease and mild cognitive impairment without dementia. AB - Impairment in delayed recall has traditionally been considered a hallmark feature of Alzheimer's disease (AD). However, vulnerability to semantic interference may reflect early manifestations of the disorder. In this study, 26 mildly demented AD patients (mild AD), 53 patients with mild cognitive impairment without dementia (MCI), and 53 normal community-dwelling elders were first presented 10 common objects that were recalled over 3 learning trials. Subjects were then presented 10 new semantically related objects followed by recall for the original targets. After controlling for the degree of overall memory impairment, mild AD patients demonstrated greater proactive but equivalent retroactive interference relative to MCI patients. Normal elderly subjects exhibited the least amount of proactive and retroactive interference effects. Recall for targets susceptible to proactive interference correctly classified 81.3% of MCI patients and 81.3% of normal elderly subjects, outperforming measures of delayed recall and rate of forgetting. Adding recognition memory scores to the model enhanced both sensitivity (84.6%) and specificity (88.5%). A combination of proactive and retroactive interference measures yielded sensitivity of 84.6% and specificity of 96.2% in differentiating mild AD patients from normal older adults. Susceptibility to proactive semantic interference may be an early cognitive feature of MCI and AD patients presenting for clinical evaluation. PMID- 14751012 TI - Working memory deficits in chronic fatigue syndrome: differentiating between speed and accuracy of information processing. AB - To examine the relative influence of speed of information processing versus working memory ability, CFS participants with psychiatric comorbidity (CFS-Psych) and CFS without a psychiatric history (CFS-noPsych) were examined on tests of visual and auditory processing speed and visual and auditory working memory. Compared to healthy controls (HC) and a group of participants with rheumatoid arthritis (RA), the CFS-noPsych group displayed significantly reduced performance on tests of information processing speed, but not on tests of working memory. No significant differences were observed between the CFS-Psych group and any other group in the study. The implications of group heterogeneity on the understanding of cognitive impairment in CFS are discussed. PMID- 14751013 TI - Hepatitis C virus infection is associated with reduced white matter N acetylaspartate in abstinent methamphetamine users. PMID- 14751014 TI - Neurological, neuropsychological, and psychosocial outcome following treatment of unruptured intracranial aneurysms: a review and commentary. AB - Thirty studies published between 1977 and 2001 that focus on outcome following unruptured intracranial aneurysm (UIA) treatment are reviewed. Although findings from these studies suggest outcome from UIA treatment is reasonably good (between 5% and 25% morbidity and between 0-7% mortality), many of the complex issues associated with the treatment of UIAs remain controversial. Most of the studies reviewed address outcome in terms of mortality and neurological morbidity. Very few studies exist which include measures of outcome such as cognitive status, psychosocial functioning and quality of life. Given that patients facing treatment tend to be healthy middle-aged adults with many years of active working and social life ahead of them, it is important to take into account the long-term consequences of either harboring an UIA, or having it treated. The small number of studies that include cognitive, psychosocial and quality of life outcomes are reviewed in some detail and suggestions made for improving future UIA outcome research. PMID- 14751015 TI - Perceptual span deficits in adults with HIV. AB - Studies have found that infection with the Human Immunodeficiency Virus (HIV) leads to cognitive dysfunction. In fact, attention problems have been reported to be the most frequent cognitive symptom in HIV-infected adults. One question is how early in the course of information processing can attention impairment be detected? To address this issue, performance on a perceptual span task was examined in 54 HIV-infected adults and 19 seronegative controls. In this task a target had to be identified in a briefly presented (50 ms) array of 1, 4, or 12 letter-characters. Response accuracy was differentially worse in the HIV+ group relative to seronegative controls in the most difficult condition, the 12-item array, but not in the easier conditions. There was no evidence of a group difference in response strategy due to disinhibition or in psychomotor speed. These data suggest that HIV infection leads to a reduction in early visual processing capacity (or span of apprehension). The present results illustrate a new type of attentional deficit in HIV and show the impact of HIV on cognition at an earlier point in information processing than has been previously reported. PMID- 14751020 TI - Healthcare influences today and tomorrow. PMID- 14751021 TI - The emerging role for health plans: "Info-Mediary". PMID- 14751022 TI - Healthcare panel: healthcare survival solutions. PMID- 14751023 TI - An employer's dilemma: grappling with health benefits. PMID- 14751024 TI - The Pharmaceutical Management Panel - highlights of the pharmacy business. PMID- 14751025 TI - Digital insurance: wave of the future? PMID- 14751026 TI - Observations on American health care. PMID- 14751027 TI - The differing strengths of Thyroid - meanderings on 2003. PMID- 14751029 TI - The effect of posttranslational modifications on the in vitro activity of recombinant human thyroid-stimulating hormone. AB - Posttranslational modification can influence the biologic activity of recombinant proteins. The effects of beta-subunit C-terminal truncation, oligosaccharide heterogeneity, and chemical oxidation on the in vitro activity of recombinant human thyroid-stimulating hormone (rhTSH) were investigated. beta-Subunit C terminal truncation up to residue 113 did not effect the in vitro activity of the hormone. The relationship between the heterogeneity of oligosaccharide structures on rhTSH and specific activity of the glycoprotein hormone was also examined. Oligosaccharide profiles were generated for preparations of rhTSH containing similar sialic acid levels. A weak correlation was observed between relative levels of monosialylated biantennary, bisialylated biantennary, and trisialylated triantennary oligosaccharide species and in vitro activity of the recombinant hormone (p < 0.05). To examine the effect of chemically induced methionine oxidation on the activity of rhTSH, the hormone was treated with tert-butyl hydroperoxide and then characterized. Using peptide mapping and mass spectrometry, the degree of oxidation of the five methionine residues within rhTSH was measured. Met-71 in the alpha-subunit was the most susceptible to oxidation whereas Met-9 in the beta-subunit was the most resistant. Also, after tert-butyl hydroperoxide treatment, levels of oxidation of Met-32 in the beta subunit, and Met-29 and Met-47 in the alpha-subunit were less than half of that observed for Met-71. The in vitro activity of rhTSH initially declined with increasing oxidation; however, the loss in activity plateaued at approximately 50% of the control sample activity. In summary, despite the possible effects that posttranslational modifications may have on the bioactivity of a protein, a limited degree of variation in bioactivity was observed for the rhTSH preparations described in this study. PMID- 14751028 TI - CTLA-4 AT-repeat polymorphism reduces the inhibitory function of CTLA-4 in Graves' disease. AB - Graves' disease (GD) is thought to be an autoimmune disease with a strong genetic component. Candidate genes include human leukocyte antigen (HLA) class II genes and CTLA-4. The CTLA-4 gene has a variable length AT-repeat polymorphism in the 3'-untranslated region. We previously found that the AT-repeat of 104 bp or longer was associated with GD. In this study, we categorized patients with GD and normal controls (NC) by genotyping the CTLA-4 AT-repeat and investigated the function of CTLA-4. Peripheral blood mononuclear cells (PBMC) and DNA were prepared from adult Caucasians (NC = 34, GD = 37). Genotypes of the AT-repeat polymorphism were divided into three groups according to their alleles. We related the CTLA-4 polymorphism in each genotype to augmentation of T-cell proliferation induced by a soluble anti-CTLA-4 antibody during incubation with irradiated Epstein-Barr virus (EBV)-transformed B cells. Proliferation of T cells from subjects with the 86/86 bp (shorter) allele was less than T cells from patients with longer alleles. The length of the AT-repeat allele correlated inversely with augmentation of proliferation after CTLA-4 blockade in subjects with GD. The CTLA-4 AT-repeat polymorphism affects the inhibitory function of CTLA-4. The long AT-repeat allele is associated with reduced control of T-cell proliferation and thus contributes to the pathogenesis of GD. PMID- 14751030 TI - Modulation of tumor necrosis factor-related apoptosis-inducing ligand-induced apoptosis by chemotherapy in thyroid cancer cell lines. AB - Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) induces apoptosis in many human cancer cells but not in normal cells. Thyroid cancer cells, however, appear to be relatively resistant to TRAIL-induced apoptosis. We therefore investigated the effect of chemotherapy on TRAIL-induced apoptosis in thyroid cancer cells. We used six thyroid cancer cell lines: TPC-1, FTC-133, FTC 236, FTC-238, XTC-1, and ARO82-1. We used flow cytometry to measure apoptosis, dimethyl-thiazol-diphenyltetrazolium bromide (MTT) assay to measure antiproliferation effects and Western blot to determine the expression of Bcl family proteins. Troglitazone, paclitaxel, geldanamycin, and cycloheximide were used for pretreatment. We used the Student's t test and analysis of variance (ANOVA) for statistical analysis. All thyroid cancer cell lines, except the TPC-1 cell line, were resistant to TRAIL, and growth inhibition was less than 20% at concentration of 800 ng/mL of TRAIL. In both TPC-1 (TRAIL-sensitive) and FTC-133 (TRAIL-resistant) thyroid cancer cell lines, pretreatment with troglitazone, cycloheximide, and paclitaxel enhanced TRAIL-induced cell death significantly but pretreatment with geldanamycin did not. There were no significant changes in Bcl 2, Bcl-xl, and Bax protein expression after troglitazone treatment. In conclusion, TRAIL in combination with troglitazone, paclitaxel, and cycloheximide induces apoptosis in thyroid cancer cells at suboptimal concentrations that cannot be achieved using TRAIL alone. PMID- 14751031 TI - Mice lacking the thyrotropin-releasing hormone gene: what do they tell us? AB - Thyrotropin-releasing hormone (TRH) is localized in the brain hypothalamus and stimulates the secretion and synthesis of pituitary thyrotropin (TSH). Although TRH deficiency caused by artificial hypothalamic destructions has been reported to result in significant decreases in TSH secretion in rodents, clinical observations from the patients with possible TRH deficiency did not entirely agree with these animal results. Because of its ubiquitous distribution throughout the brain and in the peripheral tissues, TRH has been suggested to possess a wide variety of functions in these regions. However, the neurobehavioral and peripheral actions of TRH still remains to be established. It has been, therefore, anticipated that detailed analysis of TRH-knockout mice might provide insight into the physiological significance of endogenous TRH. The present review focuses on the phenotypic findings of mice deficient in TRH. PMID- 14751032 TI - The thyrotropin receptor mutation database: update 2003. AB - In 1999 we have created a TSHR mutation database compiling TSHR mutations with their basic characteristics and associated clinical conditions (www.uni leipzig.de/innere/tshr). Since then, more than 2887 users from 36 countries have logged into the TSHR mutation database and have contributed several valuable suggestions for further improvement of the database. We now present an updated and extended version of the TSHR database to which several novel features have been introduced: 1. detailed functional characteristics on all 65 mutations (43 activating and 22 inactivating mutations) reported to date, 2. 40 pedigrees with detailed information on molecular aspects, clinical courses and treatment options in patients with gain-of-function and loss-of-function germline TSHR mutations, 3. a first compilation of site-directed mutagenesis studies, 4. references with Medline links, 5. a user friendly search tool for specific database searches, user-specific database output and 6. an administrator tool for the submission of novel TSHR mutations. The TSHR mutation database is installed as one of the locus specific HUGO mutation databases. It is listed under index TSHR 603372 (http://ariel.ucs.unimelb.edu.au/~cotton/glsdbq.htm) and can be accessed via www.uni-leipzig.de/innere/tshr. PMID- 14751033 TI - Evaluation of the therapeutic efficacy of different levothyroxine preparations in the treatment of human thyroid disease. AB - At the present time, optimal therapy for hypothyroidism requires replacement of the deficiency in thyroid hormone with synthetic levothyroxine. Precise titration of this narrow therapeutic index drug is necessary to return the patient to a chemically and clinically euthyroid state. Seven levothyroxine formulations are Food and Drug Administration (FDA)-approved and four are available to the physician. Proper dosage is established based on thyrotropin (TSH) testing and clinical evaluation. Each levothyroxine preparation must comply with FDA standards for bioavailability but may vary with respect to its dissolution and absorption properties and are not interchangeable. This equivalence testing is done on normal volunteers and requires a suprapharmacologic dose of levothyroxine in order to make the determination of bioavailability. In this review we discuss the various methods to evaluate therapeutic efficacy and bioequivalence of levothyroxine preparations in the treatment of thyroid disease. These are relevant to the physician and patient because small differences in the efficacy can produce unwanted effects of either underreplacement or overreplacement. PMID- 14751034 TI - The management of subclinical hyperthyroidism by thyroid specialists. AB - Subclinical hyperthyroidism is a relatively common condition for which prospectively derived evidenced-based management guidelines do not exist. We have conducted a case-based mail survey to solicit opinions from members of the American Thyroid Association (ATA) about various issues that arise in the management of patients with this disorder. The survey was completed and returned by 185 of 300 (62%) of the original survey recipients. Four hypothetical cases varying in age, thyrotropin (TSH) level and underlying etiology were presented. The majority of respondents recommended further evaluation of all cases, most commonly choosing a radioactive iodine uptake (42%-71%), thyroid scan (39%-68%) and antithyroid (TPO/Tg) antibodies (49%-55%) as the additional tests to be ordered. The large majority (84%) recommended observation rather than active treatment for a young patient with a low but detectable serum TSH level. A small majority also recommended observation alone for a young woman with an undetectable serum TSH level (58%) and for an older woman with a low but detectable serum TSH value (63%). However, the majority (66%) favored treating an older woman with an undetectable serum TSH. When treatment was advised in the patients with subclinical hyperthyroidism, the respondents strongly favored anti thyroid drugs when the etiology was Graves' disease and radioactive iodine when the etiology was toxic nodular thyroid disease. In the absence of adequate evidence-based guidelines, it is hoped that this survey of expert opinions may provide useful guidance for physicians providing care for patients with subclinical hyperthyroidism. PMID- 14751035 TI - Graves' dermopathy and acropachy are markers of severe Graves' ophthalmopathy. AB - It is generally considered that thyroid dermopathy and acropachy almost always occur with Graves' ophthalmopathy and that these two extrathyroidal manifestations are indicators of severe autoimmune disease and hence of more severe ophthalmopathy. However, documentation of these anecdotal impressions is needed. We assessed the presence of optic neuropathy and frequency of orbital decompression in 2 referral cohorts: 40 patients with acropachy and dermopathy (acropachy group) and 138 patients with Graves' dermopathy and no acropachy (dermopathy group). We compared those cohorts with a cohort of 114 patients who had ophthalmopathy without dermopathy and acropachy (control group). We considered optic neuropathy and the need for orbital decompression to be indicators of severe Graves' ophthalmopathy. The frequency of orbital decompression was significantly higher in the dermopathy group than in the control group (odds ratio, 3.55) and even higher in the acropachy group (odds ratios: 20.68 for acropachy group compared with control group; 5.83 for acropachy group compared with dermopathy group). The same trend occurred with optic neuropathy but was not statistically significant (alpha = 0.05; p = 0.07). Five patients were exceptions: they had definite Graves' dermopathy without clinically obvious ophthalmopathy. In conclusion, dermopathy and acropachy appear to be markers of severe ophthalmopathy. Occasionally, however, Graves' dermopathy occurs without clinical ophthalmopathy. PMID- 14751036 TI - Thyroperoxidase gene mutations in congenital goitrous hypothyroidism with total and partial iodide organification defect. AB - Mutations of the thyroperoxidase (TPO) gene have been reported as being the most severe and frequent abnormality in thyroid iodide organification defect (IOD) causing goitrous congenital hypothyroidism. The objective of this study was to screen and subsequently identify TPO gene mutations in patients with congenital hypothyroidism with evidence of total iodine organification defects (TIOD) or partial iodine organification defect (PIOD) as defined by the perchlorate discharge test. Seven goitrous patients with TIOD and seven patients with PIOD, from three and five unrelated families, respectively, were studied. We were able to detect different TPO genes mutations in patients with TIOD and PIOD. In TIOD families the results were as follows: (1) a homozygous GGCC insertion at exon 8, position 1277 (family 1); (2) compound heterozygosity with a GGCC insertion at exon 8 (1277) and a nucleotide substitution in exon 11 (2068G>C) (family 2); (3) compound heterozygosity with the mutation 2068G>C in exon 11 and a C insertion in exon 14 between positions 2505-2511 (family 3). In patients with PIOD we have detected: (1) only one heterozygous mutation in two families (4 and 5), in exons 11 and 10 (2084G>A and 1780C>A); (2) a compound heterozygous condition in one family (family 6), with mutations, respectively in exons 8 and 10 (1242G>T and 1780C>A); (3) only polymorphisms (family VII) and (4) a heterozygous mutation in the first base of the border exon/intron 9 +1G>T (family VIII). We did not detect inactivating mutations in exons 11, 16, and 21 of the THOX2 gene where mutations have been previously described. We concluded that homozygous and compound heterozygous mutations found in TIOD characterized the autosomal recessive mode of inheritance and will translate a nonfunctional protein or a protein that may not reach the apical membrane. As for PIOD, the majority of the studied kindreds had only heterozygous mutations and/or polymorphisms. It is conceivable that these TPO gene sequence alterations may partially affect the functional state of the translated protein or affect its transport to the apical membrane. PMID- 14751037 TI - The effect of thyrotropin-receptor blocking antibodies on stimulating autoantibodies from patients with Graves' disease. AB - The hyperthyroidism of Graves' disease (GD) is caused by thyrotropin-receptor (TSHR) stimulating autoantibodies (TSAb), which lead to overproduction of thyroid hormones. In this study we tried to block the stimulatory effect of patients' TSAb to the TSHR with monoclonal antibodies (mAbs) and sera from hypothyroid patients. Two groups of blocking mAbs raised by different methods from two independent groups were tested for their ability to inhibit TSH binding to the TSHR, and also the binding of TSAb from the serum of patients with GD. Group 1 mAbs (7E3, 3H10, 4C1, 1B1, 4E9) bind to amino acids 378-387 and group 2 mAbs (23.1 and 31.7) to amino acids 382-415 of the human TSHR. These results were compared to the TSH- and TSAb-inhibiting effect of sera from hypothyroid patients containing bona fide thyroid blocking antibodies (TBAb) without agonistic activity. All studies were done in a conventional cyclic adenosine monophosphate (cAMP) or a modified luciferase reporter gene bioassay. TSH-induced cAMP/luciferase signal was reduced (> 70% inhibition) by all 7 mAbs, verifying the blocking nature. Comparable results (82.2%-96.3% inhibition) were seen when cells were preincubated with 8 TBAb sera. These TBAb sera also inhibited cAMP/luciferase induction of TSAb-positive sera from patients with GD (median of 27 experiments 62.2% inhibition; range, 26.8%-93.9%), and maintained inhibition greater than 20% even when diluted 1:150. However, when mAbs were incubated with these sera, results were heterogeneous: 17 of 30 sera (57%) incubated with mAb 31.7 caused reduced cAMP production compared to incubation with the control antibody, as did 18 of 34 sera (53%) incubated with mAb 7E3, 17 of 33 sera (52%) incubated with mAb 3H10, and 16 of 31 (52%) with mAb 23.1. Mixing all four mAbs did not enhance the cAMP-reductive effect (16/27 sera; 59% inhibited). Inhibition was less pronounced than with TBAb sera (0%-76% of a control antibody) and only present at antibody concentrations greater than 10 microg/mL. We conclude that despite the strong TBAb activity of the mAbs, their effect on TSAb-induced TSHR activation of sera from patients with GD was weaker than that of human TBAb autoantibodies. Thus, the latter are not only strong inhibitors of TSH activity, but also block the stimulatory effect of autoantibodies from patients with GD. However, this effect could not be reproduced by experimental mAbs to the same extent, because it may be the result of a broader spectrum of antibodies present in the TBAb sera, interacting with or in the vicinity of TSAb epitopes. Also of interest, when a TBAb serum was added to a TSAb serum, the TBAb effect was predominant even at high dilutions. PMID- 14751038 TI - Diagnostic utility of thyroglobulin detection in fine-needle aspiration of cervical cystic metastatic lymph nodes from papillary thyroid cancer with negative cytology. AB - Cystic changes in metastatic cervical lymph nodes (CLN) from papillary thyroid cancer (PTC) may be a diagnostic pitfall in fine-needle aspiration biopsy (FNAB) cytology. We investigated in a series of CLN metastases from thyroid cancers (TC), including cystic PTC, and from a wide spectrum of extrathyroidal malignancies, the diagnostic role for metastatic TC of the rapid detection of thyroglobulin in eluates from FNAB (FNAB-Tg) of CLN. The study was carried out in a group of 79 subjects (22/57 M/F; median age, 56 years; range, 20-86 years) with enlarged CLN and thyroid nodules (TN), examined for potential metastatic TC, and harboring a large spectrum of incidentally diagnosed extrathyroidal malignancies (n = 24, mostly represented by lymphomas, lung, and breast cancers), CLN metastases from thyroid cancers (n = 28, including 6 cystic metastatic PTC), 6 specific lymphadenitis and 21 reactive lymphadenitis mostly detected (n = 16) during follow-up of patients with previously ablated TC. Markedly high FNAB thyroglobulin (Tg) values were found in all metastatic CLN TC. Two of the six cases with cystic metastatic CLN PTC were diagnosed by FNAB-Tg but not by cytology. In conclusion, FNAB-Tg has been confirmed as an easy modality and fast procedure to diagnose CLN metastasis from TC and high FNAB-Tg values with nondiagnostic cystic cytology strongly suggest cystic metastatic PTC. PMID- 14751039 TI - Impact of thyroxine-binding globulin on thyroid hormone economy during pregnancy. AB - Pregnancy-associated changes in thyroid hormone economy are well-established and are of significant clinical relevance to women with established hypothyroidism because they usually result in increased thyroxine dose requirements by these women. Studies suggest that elevations in serum thyroxine-binding globulin (TBG) have the most influence on this increased need for thyroxine, although the exact contributions by TBG rises and by other mechanisms is as yet unclear. We report the case of a 42-year-old woman, with both established primary hypothyroidism and TBG deficiency, who we have now managed through two full-term pregnancies. The patient was noted to have a baseline TBG that was approximately 30% of the average baseline level reported for non-TBG-deficient individuals. Her TBG levels were induced by pregnancy, although the absolute increase of 1.0 mg/dL was only half the increase usually associated with pregnancy. Despite the patient's low baseline TBG level and her blunted pregnancy-associated TBG induction, her absolute and relative pregnancy-associated increases in thyroxine replacement dosage mirrored those found in non-TBG-deficient, hypothyroid women. Thus, our limited study suggests that an increase in TBG concentration is not the key determinant for the increase in thyroxine requirement in pregnancy. PMID- 14751040 TI - Tablet formulation of levothyroxine is absorbed less well than powdered levothyroxine. AB - The comparative bioavailability of oral doses of levothyroxine (LT(4)) formulation taken as tablets, after being crushed, or chewed before swallowing has not been well studied. Three patients with hypothyroidism who showed persistent elevation of serum thyrotropin (TSH) despite taking 200, 150, and 125 microg of LT(4) tablets per day are presented. They did not show signs and symptoms of gastrointestinal illness that could interfere with the absorption of LT(4) nor history of such a condition. They did not concurrently take medications known to affect the absorption of LT(4) from the gut. Their serum TSH levels normalized when the tablets were taken after being pulverized. The difference appeared attributable to the slow dissolution of the tablets in the gut of these patients. PMID- 14751041 TI - Combined positron emission tomography - computed tomography improves specificity for thyroid carcinoma by identifying vocal cord activity after laryngeal nerve paralysis. PMID- 14751042 TI - Three unusual causes of diffuse thyroid enlargement. PMID- 14751043 TI - Autoimmune thyroiditis after elimination of iodine deficiency in Sri Lanka. PMID- 14751044 TI - Hashimoto's and Ord's diseases. PMID- 14751047 TI - [Effect of human cytomegalovirus on hematopoietic system]. AB - OBJECTIVE: To investigate the mechanism and the suppression effect of human cytomegalovirus (HCMV) on hematopoietic system. METHODS: Semi-solid culture system was used to observe the effect of HCMV AD169 strain on colony forming unit granulocyte/macrophage (CFU-GM), CFU-erythroid (CFU-E), CFU-multipotent (CFU-Mix) and CFU-megakaryocyte (CFU-MK) growth. The techniques of in situ polymerase chain reaction (IS-PCR) and polymerase chain reaction (PCR) were used to demonstrate the existence of HCMV DNA in the colony cells of cultured CFU-GM, CFU-Mix, CFU-MK and CFU-E, respectively. The immediate early antigen (IEA) mRNA in CFU-MK and late antigen (LA) mRNA in CFU-E were detected by reverse transcriptase-polymerase chain reaction (RT-PCR). HCMV early protein P52 was detected with immunohistochemical technique. RESULTS: HCMV AD169 suppressed the differentiation and proliferation of CFU-GM, CFU-E, CFU-Mix and CFU-MK in vitro significantly (P < 0.05). The suppression was dose-dependent. HCMV DNA was successfully detected in CFU-GM, CFU-Mix, CFU-MK colony cells from viral infection groups by IS-PCR, and was detected in CFU-E by PCR, while it was negative in blank control or mock control groups. CFU-MK colony cells expressed HCMV IEA mRNA with the size of 340 bp in virus infection groups of 10(3) plague forming unit (PFU), 10(4) PFU and 10(5) PFU, respectively. The HCMV LA mRNA was detected by RT-PCR and was 263 bp long in positive control group of HCMV-infected human embryonic fibroblasts. The expression of HCMV LA mRNA in CFU-E was negative. The early protein P52 of HCMV in 10(4) PFU group was also identified by immunohistochemical staining. CONCLUSION: HCMV AD169 strains inhibited the differentiation and proliferation of CFU-GM, CFU-E, CFU-Mix and CFU-MK by the infection of the hematopoietic progenitors. HCMV might cause the suppression of hematopoiesis by direct infection, which is thought to be one of the reasons of HCMV infection associated with thrombocytopenia, neutropenia and anemia. PMID- 14751048 TI - [Analysis of 21 children with acute non-lymphoid leukemia carrying AML1/ETO fusion gene]. AB - OBJECTIVE: It was revealed that t(8; 21) (q22; q22) was one of the most common chromosomal aberrations in acute non-lymphoid leukemia. The translocation was found to be involved in the AML1 gene on the chromosome 21 and the ETO gene on the chromosome 8, and resulted in the formation of AML1/ETO fusion gene on the derivative chromosome 8. The fusion gene was a transcription factor and played a direct role in the leukemogenesis. The translocation was mainly observed in M(2), accidentally in M(4) and M(1) and rarely in MDS. Here we studied the main clinical data in children with acute non-lymphoid leukemia (ANLL) carrying the AML1/ETO fusion gene. In addition, we discussed the significance of the detection of AML1/ETO fusion gene in the diagnosis and prognosis of children with ANLL. METHODS: The authors investigated 29 patients in our hospital from December 2000 to March 2002. The patients were divided into two groups. Group A included 21 patients, 14 males and 7 females. They were 3.6 to 14 years old and the median was 9. Group B included 8 patients, 6 males and 2 females. They were 0.8 to 14 years old and the median was 6. Diagnosis was made according to FAB and MIC criteria and the expression of AML1/ETO fusion gene was detected with nested RT PCR. The patients were treated according to DA, DAE or BFM regimen, respectively. The main clinical indexes including age, Hb, white blood count, platelet, blasts in PBC and BM, and time of arrival at complete remission (CR), were compared statistically between the two groups with t test of independent samples. RESULTS: All the 21 patients in group A were found carrying AML1/ETO, and 17 patients (81%) were classified as M(2), the other 4 cases were of M(2) developed from MDS RAEB-T, M(4Eo), M(5) and eosinophil leukemia, respectively. Eighteen out of 20 patients whose effects could be assessed reached CR, and the CR ratio was 90%. Two patients in group B were of AML-M(1), 3 M(2), 1 M(3), 1 M(4), and 1 M(5), respectively. None of them was found carrying AML1/ETO. Seven cases reached CR and the ratio was 87.5%. There was no significant difference between the two groups in the above clinical indices. CONCLUSIONS: Between the two groups of patients there was no significant difference in the above clinical indices. RT PCR for the detection of AML1/ETO in children with ANLL was quick, convenient and sensitive, and could be regarded as a useful method for the diagnosis and prognosis of ANLL. PMID- 14751049 TI - [Effect of truncated platelet-derived growth factor-alpha receptor on apoptosis and expression of c-sis mRNA of pulmonary artery smooth muscle cells]. AB - OBJECTIVE: Platelet-derived growth factor (PDGF) plays an important role during the pathophysiological changes in vascular remodeling. The study aimed to investigate the effect of truncated PDGF-alpha receptor on apoptosis and expression of c-sis mRNA of pulmonary artery smooth muscle cells (VSMCs). METHODS: Tissue mass culture was done to get vascular smooth muscle cells of pulmonary artery in newborn pigs. Two methods were used to interfere VSMCs: adding adenoviral recombined body (Ad5CMV-PalphaRtr, ACP) with three different concentrations of truncated PDGF-alpha receptor into the cultures, or adding three concentrations of PDGF-BB after the treatment with mid-concentration of ACP. VSMC apoptosis, cellular cycle and expression of c-sis were observed using flow-cytometry, and the expression of c-sis mRNA was detected by reverse transcription polymerase chain reaction (RT-PCR). RESULTS: ACP with mid- to- high concentrations could restrain the proliferation of VSMCs apparently with the increase of G(0)/G(1) cells. The apoptotic rate presented an ascending tendency. The differences among the groups were of statistically significant. Affected by mid- concentration of ACP, PDGF-BB did not exhibit a significantly accelerating effect on the changes of cellular cycle and VSMC apoptosis. The expression of c sis mRNA was up-regulated under the effect of ACP. Affected by mid-concentration of ACP and PDGF-BB, c-sis mRNA expressed was down-regulated. CONCLUSION: Mid- to- high concentration of ACP is a powerful inhibitor of cellular proliferation for pulmonary artery VSMCs. It can significantly increase cells in number in G(0)/G(1) phase, apoptosis and c-sis mRNA expression. PMID- 14751050 TI - [Immunophenotyping of 222 children with acute leukemia by multi-color flow cytometry]. AB - OBJECTIVE: Acute leukemia (AL) is one of the most common malignant diseases in children. AL immunophenotypes are known to be benefit to the predictable prognoses and specific therapy. Recently, the accuracy of AL immunophenotyping was dramatically improved with the application of the flow cytometry, the new monoclonal antibodies, the improvement of the gating strategies and the multi parameter analytic techniques. The aim of this study was to evaluate the value of multi-color flow cytometry in the immunophenotyping of acute leukemia in children. METHODS: Three- or four-color flow cytometry and CD(45)/Side Scatter (SSC) gating were used to analyze the surface and cytoplasmic (Cy) antigen expressions in 222 successive cases of childhood acute leukemia. RESULTS: Cells from 222 cases of children with acute leukemia were analyzed. Based on the diagnostic criterion proposed by European Group for the Immunological Characterization of Leukemia (EGIL), four categories could be identified: the undifferentiated type accounted for 0.9%, acute myeloid leukemia (AML) 35.1%, acute lymphoblastic leukemia (ALL) 55.9%, and mixed lineage AL 8.1%. Of 124 patients with ALL, 94 patients (75.8%) were classified as B lineage and 30 patients (24.2%) T lineage ALL. Antigen aberrant expressions were found in AML (24.4%), B lineage ALL (36.2%) and T lineage ALL (30.0%). CD(7) was the most commonly expressed lymphoid antigen in AML (12.8%), followed by CD(19) (6.4%) and CD(2) (5.1%). CD(13) was the most commonly expressed myeloid antigen in ALL (18.5%), followed by CD(15) (11.3%), CD(11b) (6.5%) and CD(33) (4.3%). CD(117) and CD(56) presented in 73.3% and 38.0% cases of AML, respectively, but were generally absent on blast cells of ALL. CyCD(22), CyCD(3) and CyMPO were specifically expressed in B lineage, T lineage and myeloid lineage leukemia, respectively, and the first two could be more sensitively detected than they were on cell membrane surface. CONCLUSIONS: Multi-color flow cytometry is a reliable technique in the diagnosis, differential diagnosis and classification of acute leukemia in children. PMID- 14751051 TI - [Evaluation of physiological index on treadmill exercise testing of 294 healthy children in Shanghai area]. AB - OBJECTIVE: Dynamic exercise is often used to evaluate the backlog function of cardiovascular system, and the treadmill test is a commonly used dynamic exercise protocol. The study aimed to assess the exercise capacity and cardiovascular response to treadmill exercise in healthy children, and create normal reference values of exercise testing in native children. METHODS: Two huadveds and ninety four healthy children aged 5 to 14 years were tested using the Bruce protocol on treadmill. RESULTS: Mean exercise capacity in boys increased from 12.2 METs at the age of 5 to 6 years, to 15.2 METs at the age of 13 to 14 years. Mean exercise capacity in girls increased from 11.7 METs at the age of 5 to 6 years, to 12.6 METs at the age of 13 to 14 years. Sex difference was obvious in exercise capacity of children except those at the age of 5 to 6 years. There was no obvious correlation between exercise capacity and the ratio of weight to height. Maximal heart rate ranged from 187 to 235 beats/min. Systolic blood pressure increased gradually until peak exercise was achieved. The mean systolic blood pressure of boys at maximal exercise increased by 38.6 percent compared to resting level, and the girls increased by 34.4 percent. After maximal exercise, mean systolic blood pressure reached resting level in 6 minutes but diastolic blood pressure varied. All children had sinus rhythm at rest. No arrhythmia was recorded during treadmill exercise. However, arrhythmia was found in five children in early recovery period. The incidence of exercise-induced arrhythmia was 1.7 percent. CONCLUSION: Sex difference and age difference was obvious in exercise capacity. In the 21st century, the exercise capacity of children in Shanghai area has approached to the developed country. Using MET as the standardized criterion of exercise capacity is advantageous to standardize maximal or submaximal exercise workloads of all kinds of protocols. Exercise induced arrhythmia in the early recovery period in children without structural heart disease may not be pathological, but it is necessary for those children to be followed-up. PMID- 14751052 TI - [Premature, low birth weight, small for gestational age and childhood cerebral palsy]. AB - OBJECTIVE: To investigate the relationship between perinatal risk factors such as premature, low birth weight, small for gestational age and childhood cerebral palsy (CP). METHODS: A cross sectional survey was carried out among 305,263 children aged 1 - 6 years old in seven cities of Jiangsu Province, China from May to July 1997. The perinatal risk factors were analysed. RESULTS: Four hundred and eighty-four cases of CP were found among this population. The prevalence of CP for children aged 1 - 6 years old was 1.59 per thousand. The prevalence of CP were strongly correlated to prematurity (RR = 25.16), low birth weight (RR = 19.63), and also highly correlated to small for gestational age (RR = 4.34). For smaller groups divided by small for gestational age (SGA), appropriate for gestational age (AGA), large for gestational age (LGA) and then by gestational age, prematurity was found to be at high risk in SGA (RR = 9.29), AGA (RR = 28.34) and LGA (RR = 21.41) groups. For groups divided by gestational age and then by SGA, AGA and LGA, SGA was found to have significantly high risk in premature (RR = 1.45), mature (RR = 4.41) and postmature (RR = 3.19) groups. Nine groups were divided by the gestational age along with SGA, AGA and LGA, rates of CP were found to be significantly higher in most groups than in the term AGA group. Compared with the rate of CP in the term AGA group, the RR were calculated and showed as followings (from higher to lower), premature SGA (RR = 40.99), premature AGA (RR = 28.34), premature LGA (RR = 21.08), postmature SGA (RR = 8.39), mature SGA (RR = 4.41) and postmature AGA (RR = 2.63). CONCLUSION: Prematurity and small for gestational age are both independent risk factors for cerebral palsy. Postmaturity and large for gestational age are not risk factors. PMID- 14751053 TI - [Expression of neutrophil adhesion molecule CD11b as an early diagnostic marker for neonatal sepsis]. AB - OBJECTIVE: Neonatal sepsis is a common disease and the sepsis-related mortality rate is still high. Until now, there has no ideal diagnostic marker to early identify neonatal sepsis. Expression of neutrophil adhesion molecule CD(11b) was showed as the earlier reaction to the infection/inflammation, and may be applied as an early diagnostic marker for sepsis. This study was to investigate this antigen for early diagnosis of neonatal sepsis related to bacterial infection. METHODS: According to clinical symptoms, signs and four indices (WBC, PLT, plasma CRP and ratio of I/T), fifty-one neonates with established or suspected sepsis were allocated retrospectively into two groups of sepsis [n = 23, gestational age of (38.3 +/- 2.4) weeks, postnatal age of (12.7 +/- 8.8) days, body weight: (3.1 +/- 0.8) kg] and suspected sepsis [n = 28, gestational age of (38.8 +/- 1.6) weeks, postnatal age of (11.7 +/- 7.3) days, body weight: (3.3 +/- 0.6) kg]. Fifteen healthy neonates were served as controls [gestational age: (38.5 +/- 1.4) weeks, postnatal age: (8.2 +/- 5.5) days, body weight: (3.3 +/- 0.3) kg]. CD(11b) was quantified with the whole blood flow cytometry and direct immunofluorescence technique. RESULTS: The expressions of neutrophil CD(11b) in neonates with sepsis and suspected sepsis were (320 +/- 189) MFI and (456 +/- 213) MFI, respectively, which was lower than that of controls [(1,090 +/- 338) MFI, t = -9.01 and -7.56, respectively; P < 0.001]. The expression of CD(11b) was lower in neonates with sepsis than that with suspected sepsis (t = -2.39, P < 0.05). The expression of CD(11b) in neonates with CRP >or= 30 mg/L was (211 +/- 164) MFI, which was lower than those with CRP < 30 mg/L [(505 +/- 265) MFI, t = 2.64, P < 0.05]. The detection of CD(11b) (60 years (n = 40), 44% versus 20%. The response rate in patients with extra pelvic disease (n = 42) was 31% as compared to 14% in those with strictly pelvic and/or vaginal disease (n = 14). The median progression-free survival (PFS) was 2.7 months and median overall survival was 14.0 months. Two patients experienced a grade 4 thromboembolic event. Additional toxicities included one of each grade 3 gastrointestinal, grade 3 neurologic, and grade 3 genitourinary. CONCLUSION: A regimen of alternating megestrol acetate and tamoxifen is active in treating endometrial cancer and may result in a prolonged complete response (CR) in some patients. PMID- 14751132 TI - Establishment of a HPV and p53-mutation-negative human cell line (CA) derived from a squamous carcinoma of the uterine cervix. AB - OBJECTIVE: Well-characterized human cancer cell lines are important research resources for studying cancer cell biology, as well as for developing new strategies against cancer cell growth and progression. We present a new cell line, CA, established from an invasive non-keratinizing squamous cell carcinoma of the uterine cervix in 36-year-old patient. METHODS: We measured the doubling time of CA cells. To investigate the tumorigenicity of CA, cells were inoculated subcutaneously into the back of nude mice. Several tumor markers were analyzed using culture media by EIA. PCR-based analyses were performed to examine the human papillomavirus (HPV) status and telomerase activity. CA was also screened for p53 mutation using the sequencing technique. RESULTS: The cells show rapid growth in culture with a doubling time of 14.3 h and high migration activity. Monolayer-cultured cells were polygonal, showing a pavement-like arrangement and a tendency to pile up without contact inhibition. Subcutaneous transplantation of the CA cells into nude mice formed solid tumors that were histologically diagnosed as squamous cell carcinoma, whereas no metastasis was observed. Cultured CA cells produced SCC, CEA, TPA, CA125 and SLX. Genetic and molecular analyses revealed high telomerase activity and the absence of HPV DNA. No p53 mutation was observed in this cell line. CONCLUSION: These properties suggest that CA is an aggressive cervical carcinoma cell line and may serve as a useful experimental model for studying HPV role in cervical carcinogenesis. PMID- 14751133 TI - Lung cancer metastatic to a cervical polyp. AB - BACKGROUND: We describe an unusual finding of a lung cancer metastatic to a cervical polyp. CASE: A 69-year-old female with metastatic lung cancer was referred for evaluation of an asymptomatic cervical polyp. The polyp was removed and pathologic examination revealed a focus of invasive adenocarcinoma with signet ring features within a polypoid fragment of squamous mucosa. All studies suggest a primary pulmonary origin. CONCLUSION: Cervical polyps occur in up to 5% of women and 1.7% contain carcinomatous changes. Most malignant polyps result from the progression of localized dysplasia, but distant metastases have been reported. We were unable to find any prior report of a primary lung cancer metastatic to a cervical polyp. Although removal or biopsy of asymptomatic polyps is reasonable, further studies need to be done. PMID- 14751134 TI - Radiation fields in gynecologic oncology: correlation of soft tissue (surgical) to radiologic landmarks. AB - OBJECTIVES: (1). To determine if radiation fields defined by bony structure landmarks correlate to anatomic boundaries of lymph node dissection marked intraoperatively; and (2). to determine if a patient's body mass index (BMI) correlates with these anatomic or radiographic boundaries. METHODS: One hundred patients undergoing exploratory laparotomy with pelvic and paraaortic lymph node dissection had three medium hemoclips placed at vascular junctions considered of clinical significance to lymph node dissection: insertion of the left ovarian vein into the renal vein, insertion of the right ovarian vein into the vena cava, inferior mesenteric artery (IMA), bifurcation of the aorta, bifurcation of the common iliacs (bilateral), and the insertion of the deep circumflex vein (DCV) in to the external iliac vein (bilateral). Postoperatively, an abdominal X-ray was obtained. Comparisons were made between these eight major vascular landmarks and radiographic bony landmarks that are used to define radiation field boundaries. The percentage of vascular landmarks that were encompassed or fell outside of traditional radiation fields was determined with a 1-cm margin considered an adequate boundary for radiation. These measurements were also compared to patient BMIs. RESULTS: Radiation fields defined by traditional bony landmarks would adequately encompass the paraaortic lymph nodes in the majority of patients (91%). For pelvic radiation fields, there was a significant "miss" (39%) of common iliac lymph nodes. Approximately one quarter (26%) of patients would receive inadequate coverage of one or both of the lateral boundaries of pelvic radiation. There was no apparent correlation of BMI to vascular or bony landmarks. CONCLUSIONS: Radiation fields determined by traditional bony landmarks do not adequately reflect the anatomic (surgical) landmarks associated with the lymphatic drainage of the female reproductive organs. Although the majority of tertiary care centers now use advanced imaging techniques (e.g. computed tomography) to plan their radiation treatments, the historical guidelines of radiographic landmarks are still used in smaller institutions and continue to be referenced in Gynecologic Oncology Group protocols. For centers still using radiographic landmarks, the application of hemoclips with X-ray identification is a low-cost modality that is easily reproducible and may be clinically useful in guiding treatment. PMID- 14751135 TI - ERBB2 amplification is superior to protein expression status in predicting patient outcome in serous ovarian carcinoma. AB - OBJECTIVE: The objective of this study was to evaluate the frequency and clinical significance of ERBB2 gene amplification in serous ovarian carcinoma. In addition, concordance of the findings of ERBB2 immunohistochemistry and ERBB2 amplification was assessed. METHODS: Tissue microarray constructed of 401 serous ovarian carcinomas was examined by chromogenic in situ hybridization (CISH) using probe for ERBB2 gene and by immunohistochemistry using CB11 monoclonal antibody against ERBB2 protein. RESULTS: Amplification (>5 copies per cell) of ERBB2 was detected in 7% and low copy number increase (three-five copies) in 14% of the carcinomas. Increased copy number of ERBB2 was associated with poor prognosis, that is, poor response to therapy (P = 0.024), shorter disease-free (P < 0.0001) and overall survival (P < 0.0001). ERBB2 copy number status was identified as an independent prognostic factor for overall survival. Increased copy number of ERBB2 was also associated with high tumor grade, greater patient age, large residual tumor size, high proliferation index, aberrant p53 and negative progesterone receptor status. A significant association was found between ERBB2 amplification and ERBB2 protein overexpression. However, a substantial number of cases showed discrepant results by the two methods, especially in cases with low level amplification or moderate protein overexpression. Overexpression of ERBB2 protein was associated with poor overall survival, but the prognostic value was weaker than that of ERBB2 gene copy number status. CONCLUSION: ERBB2 amplification positive tumors identified by CISH constitute a subgroup of serous ovarian carcinomas associated with aberrant p53, negative progesterone receptor status and aggressive behavior, a suitable group for testing the effect of trastuzumab in clinical trials. PMID- 14751136 TI - Cyclins and proliferation markers in early squamous cervical carcinoma. AB - OBJECTIVES: To examine the prognostic significance of the protein expression of cyclin E, cyclin A and cyclin D3, and of the proliferation markers Ki-67 and BM28 in early squamous cervical carcinoma (SCC). METHODS: Tissue blocks from 221 patients who underwent radical hysterectomy and bilateral lymphadenectomy at our institution for stage IB SCC between 1987 and 1993 were available for this study. Immunohistochemistry using monoclonal antibodies against cyclin E, cyclin A, cyclin D3, Ki-67 and BM28 was used to examine protein expression. Ten patients who underwent hysterectomy for uterine prolapse served as controls. RESULTS: Cyclin E, cyclin A, Ki-67 and BM28 expression was increased in SCC and high expression was observed in 81.5% (180/221), 35% (78/221), 25.5% (56/221) and 92% (204/221) of tumors, respectively. Cyclin D3 was decreased in SCC and low expression was found in 50.5% (111/220) of tumors. In univariate analysis, all classical clinicopathological parameters but none of the investigated proteins were associated with prognosis. In multivariate analysis, only deep stromal invasion was independently related to survival. CONCLUSION: Cyclin E, cyclin A and cyclin D3 and the proliferation markers Ki-67 and BM28 are not independently associated with prognosis in stage IB SCC. PMID- 14751137 TI - Tumor-stromal cell contact promotes invasion of human uterine cervical carcinoma cells by augmenting the expression and activation of stromal matrix metalloproteinases. AB - OBJECTIVE: The augmentation of the expression and activation of matrix metalloproteinases (MMPs) is associated with tumor invasion and metastasis. In addition, tumor-stromal cell contact provides a crucial signal for regulating the pericellular proteolysis for the progression of tumor invasiveness. The present study evaluates the regulation of the expression and activation of MMPs and tissue inhibitors of metalloproteinases (TIMPs) by tumor-stromal cell contact in an in vitro co-culture model of human uterine cervical carcinoma cells and human uterine cervical fibroblasts. METHODS: When human uterine cervical carcinoma SKG II cells were co-cultured with human uterine cervical fibroblasts (HUCFs), the invasive activity of SKG-II cells was analyzed using an in vitro invasion assay using Matrigel. The production, mRNA expression and activation of MMPs and TIMPs were monitored by Western blot and Northern blot analyses and gelatin zymography. RESULTS: SKG-II cells, which constitutively produced membrane-type 1 MMP (MT1 MMP) and a trace of proMMP-2 but neither TIMP-1 nor TIMP-2, showed poor invasiveness in vitro. Upon co-culturing with HUCFs, SKG-II cells were found to transform to the invasive phenotype by enhancing the production and mRNA expression of tumoral MT1-MMP. In addition, a sequential increase in the activation of fibroblast proMMP-2 was observed along with the formation of an MT1 MMP-TIMP-2-proMMP-2 complex on the tumor cell surface. Furthermore, the production and gene expression of fibroblast proMMP-1 and proMMP-3 were augmented under co-culture conditions, whereas mRNA expression of proMMP-2, TIMP-1 and TIMP 2 was unchanged. Moreover, we demonstrated the partial involvement of tumor-cell derived soluble factors in the augmentation of the production of proMMP-1 and proMMP-3 in HUCFs. However, anti-integrin beta1 and beta3 antibodies failed to abolish the augmentation of fibroblast proMMP-3 production and proMMP-2 activation in the co-culture. CONCLUSION: Cell-cell contact between cervical carcinoma cells and peripheral stromal fibroblasts augments the production and activation of MMPs, and therefore the subsequent imbalance between MMPs and TIMPs may result in the progression of invasiveness of cervical carcinoma cells in vivo. PMID- 14751138 TI - Poly(L-lactide/glycolide) suture: the effect of acute radiation. AB - The physical properties of Poly(L-lactide/glycolide) indicate that the suture retains approximately 80% of its original strength at 3 months and 60% of its original strength at 6 months. This new long-term synthetic absorbable suture offers postoperative fascial strength for an extended period when compared to other available absorbables. Importantly, many women with gynecologic cancer will undergo radiation therapy during this interval. This report is intended to evaluate the effects of ionizing radiation on this suture. METHODS: One lot (op strands) of size 1 PLG suture was used. Ten (10) strands were used for baseline study of out of package tensile strength. Eighty (80) strands were placed in a buffer solution (pH approximately 9.0) and incubated in an in vitro water bath approximately 48 h at 55 degrees C to simulate 4 weeks in vitro residence time. Following this in vitro hydrolysis 20 strands were tested. Sixty strands were placed beneath a piece of fresh full thickness porcine skin and subcutaneous tissue and exposed to 3, 30, and 70 Gy. All samples were evaluated for breaking strength and elongation-at-break using an Instron tensiometer. RESULTS: Analysis of variance performed at the different exposure level revealed no significant effect on tensile properties (p>0.1). CONCLUSION: The tensile properties of PLG suture are not adversely affected by ionizing radiation. This long-term absorbable suture is an alternative for fascial closure when extended periods of support are necessary in patients destined to receive therapeutic ionizing radiation. PMID- 14751139 TI - Phase II trial of neoadjuvant paclitaxel and cisplatin in uterine cervical cancer. AB - OBJECTIVES: The toxicity and effectivity of intravenous paclitaxel and cisplatin as neoadjuvant chemotherapy were assessed in cervical cancer patients. Patients and methods. Forty-three consecutive patients affected by International Federation of Gynecology and Obstetrics (FIGO) stage IB2 to IIB were treated with paclitaxel 60 mg/m(2) that was administered intravenously over a 3-h period, followed by cisplatin 60 mg/m(2), also administered intravenously. The chemotherapy was administered every 10 days and for three courses. The toxicity of the regimen in each cycle was determined according to the WHO toxicity criteria and in cases with grade 3 or 4 toxicity, chemotherapy was postponed until the toxicity was disappeared. Before the neoadjuvant chemotherapy, the lesion was pathologically confirmed by punch biopsy, and the size of the tumor mass was measured by pelvic examination and pelvic magnetic resonance imaging (MRI). The response to the treatment was determined 10 days after three cycles of chemotherapy by pelvic examination and pelvic MRI. Two weeks after the neoadjuvant chemotherapy was completed, the patients were either given an operation or radiation therapy, depending on their overall condition, the operational risks, and personal willingness for an operation. RESULTS: A total of 43 patients were enrolled in this study and all of them were given an operation. Hematologic toxicity was seen in 17 patients. But most of them were anemia and there was no grade 3 or 4. Grade 1 neurotoxicities developed in 29 patients and all of them were peripheral neurotoxicity. Clinical responses occurred in 90.7% (39/43) of patients, including 39.5% (17/43) with a complete response (CR), 11.6% (5/43) with a pathologically determined complete response, 51.2% (22/43) with a partial response (PR), and 9.3% (4/43) showed stable disease (ST). A down-staging response was seen in 72.1% (31/43) of those patients showing a response. CONCLUSION: The combination of paclitaxel with cisplatin for use in neoadjuvnant chemotherapy seems to be tolerated and very active in cervical cancer. Especially, every 10 days treatment did not delay the optimal time of main treatment. A larger number of cases need to be studied to confirm the efficacy of the treatment. PMID- 14751140 TI - Clinicopathologic features of villoglandular papillary adenocarcinoma of the uterine cervix. AB - OBJECTIVES: The objectives of this study are to analyze the clinicopathologic features of villoglandular papillary adenocarcinoma (VGPA) of the uterine cervix and to discuss the management thereof. We examined 13 patients with VGPA. METHODS: Clinical profiles, including patient age, clinical stage, surgical procedure, and outcome, were recorded. Pathologically, macroscopic features, polypoid tumor size, horizontal spread and depth of endophytic tumor, nuclear atypicality, mitotic count, lymph capillary space invasion, and lymph node metastasis were investigated. RESULTS: The median age of 13 patients was 45 years, with 10 and 3 patients staged Ib and IIb, respectively. All the patients underwent hysterectomy and pelvic lymphadenectomy and are alive without recurrence. Macroscopically, the tumor showed a polypoid pattern in 8 patients and a flat pattern in the remaining 5 patients. Polypoid tumor size ranged between 4 x 2 and 20 x 15 mm. Horizontal spread and depth of endophytic tumor ranged between 8 and 30 mm and between 3 and 11 mm, respectively. The tumor in all the 13 patients except 1 showed moderate nuclear atypicality. The mean mitotic count was 43/10 high-power fields. Lymph capillary space invasion was present in 4 patients, 1 of whom also had bulky lymph node metastases. CONCLUSIONS: VGPA has been reported to rarely involve lymph capillary space invasion or lymph node metastasis, leading some surgeons to conduct less radical surgeries such as conization. Nevertheless, we encountered patients with these pathologic risk factors. Much caution should be exercised in managing patients with VGPA. PMID- 14751141 TI - Immunohistochemical markers in differential diagnosis of endometrial stromal sarcoma and cellular leiomyoma. AB - OBJECTIVE: Distinction of endometrial stromal sarcoma (ESS) from benign smooth muscle proliferations like cellular leiomyoma (CL) is sometimes problematic. The purpose of this study was to evaluate the potential utility of a panel of antibodies in the differential diagnosis of ESS and CL. METHODS: Using a standard streptavidin-biotin method, the expression of desmin, alpha smooth muscle actin (SMA), calponin h1, h-caldesmon, estrogen receptor (ER), progesterone receptor (PR), CD10, CD44v3, proliferating cell nuclear antigen (PCNA), and mast cells (MCs) were evaluated in 26 cases of ESS (21 low grade, 5 high grade), 25 CL (17 common CL, 8 highly CL), 25 myometria, and 25 endometria. RESULTS: Among ESS, 20 of 26, 17 of 26, 9 of 26, 12 of 26, 14 of 26, and 22 of 26 were positive for expression of desmin, SMA, calponin h1, ER, PR, and CD10, respectively, while only 2 of 26 were positive for CD44v3 and all were entirely negative for h caldesmon. Of CL, all were positive for SMA, calponin h1, PR, and CD44v3; 24 of 25, 24 of 25, and 19 of 25 were positive for desmin, h-caldesmon, and ER, respectively, whereas 1 of 25 focally marked with antibodies to CD10. There was no significant difference of PCNA expression between ESS and CL, although the ESS cases tended to have higher values. The MC counts were significantly higher in the CL group than in the ESS group (P < 0.01). When using the cut-off value of seven MCs per HPF to distinguish ESSs from CLs, the sensitivity and specificity of this cut-off value were 92.9% and 100%, respectively. CONCLUSIONS: A panel of h-caldesmon, CD10, and CD44v3 should be used and will distinguish ESS from CL in most cases. In addition, counting the number of MCs might be useful as part of a multivariate approach to the differential diagnosis of them. But the biological function of MC and CD44v3 in these tumors is worthy of further investigation. PMID- 14751142 TI - Plasminogen fragmentation and increased production of extracellular matrix degrading proteinases are associated with serous epithelial ovarian cancer progression. AB - OBJECTIVE: Elevated levels of proteases are linked to the malignant phenotype in a wide variety of solid tumors. Therefore, the expression of plasminogen, matrix metalloproteinases (MMP-2 and MMP-9), and of the serine protease urokinase-type plasminogen activator (uPA) in serous epithelial carcinoma of the ovary were investigated. METHODS: Plasminogen antigen was analyzed in tissue extracts and in the urine of patients with normal (n = 12), benign (n = 6), borderline (n = 9), and invasive serous tumors (n = 22) by Western immunoblotting using rabbit polyclonal plasminogen and murine monoclonal angiostatin antibodies. In the same tissue extracts, semiquantitative estimates of MMP-2, MMP-9, total MMP activity, and uPA activity were determined using semiquantitative gelatin zymography in the presence or absence of human plasminogen. RESULTS: Bands corresponding to Glu plasminogen (approximately 92 kDa) and Lys-plasminogen (approximately 86 kDa) were detected in all ovarian tissues and in corresponding urine samples. Densitometric analysis of combined Glu- or Lys-plasminogen levels showed significantly decreased levels in malignant compared to normal tissue. In Grade 3 cancers, there was no evidence of Glu-plasminogen or angiostatin. MMP activity was significantly elevated in both borderline and in Grade 3 ovarian cancer tissues. Increased tissue uPA activity on zymograms was detected only in Grade 3 ovarian cancer tissue. CONCLUSION: These data suggest that proteolytic activity of the plasminogen activation cascade increases in serous epithelial ovarian carcinoma. PMID- 14751143 TI - Sentinel node detection in patients with vaginal carcinoma. AB - OBJECTIVE: The objective was to describe sentinel ode detection in patients with primary and recurrent vaginal carcinoma. METHOD: Preoperatively 60-mBq technetium labeled nannocolloid was injected in the mucosa at 3, 6, 9, and 12 o'clock, just adjacent to the vaginal cancer. Sentinel nodes were detected using a laparoscopic or hand-held probe (Navigator) and removed for pathological assessment. RESULTS: Sentinel nodes could be found in two of three patients with primary stage I or II carcinoma of the vagina. In the first patient the sentinel nodes, located in the groin and obturator region, proved to be negative and she was treated with a wide local excision of the tumor, pelvic and groin lymphadenectomy, and adjuvant radiotherapy. The second patient had tumor metastases in the sentinel node, which was found just below of the junction of the iliac vessels, and she underwent combined chemo- and radiotherapy. In a third patient no sentinel node could be detected at lymphoscintigraphy. A last patient with stage III carcinoma of the upper vagina was initially treated by combined chemo-radiotherapy but recurred 6 months later. During a staging procedure the sentinel nodes could be detected in the right obturator fossa and were removed laparoscopically. As they were negative, she underwent a posterior pelvic exenteration with complete resection of the tumor. CONCLUSION: Laparoscopic detection of sentinel nodes using 99mTc labeled colloid is feasible in patients with primary and recurrent vaginal cancer and may provide important information to direct further management. PMID- 14751144 TI - Phase I feasibility trial of carboplatin, paclitaxel, and gemcitabine in patients with previously untreated epithelial ovarian or primary peritoneal cancer: a Gynecologic Oncology Group study. AB - PURPOSE: To determine the feasibility of administering a minimum of four cycles of carboplatin, paclitaxel, and gemcitabine (CPG) every 21 days without excessive dose modification or cycle delay in patients with previously untreated epithelial ovarian cancer or primary peritoneal cancer. METHODS: Paclitaxel 175 mg/m(2) was given over 3 h followed by carboplatin concentration time curve (AUC) 5 (day 1) and gemcitabine 1 g/m(2) (days 1 and 8) in the first cohort. A second cohort received paclitaxel 135 mg/m(2) over 3 h followed by carboplatin AUC 5 (day 1) and gemcitabine 800 mg/m(2) (days 1 and 8). A maximum of eight cycles was administered. RESULTS: Fourteen patients received 89 cycles during the first cohort. Seven patients experienced 19 hematologic dose-limiting events (DLEs) within the first four cycles, including grade 4 thrombocytopenia (n = 9), febrile neutropenia (n = 3), and omission of gemcitabine on day 8 (n = 7). This exceeded the threshold for nonfeasibility. In the second, less intense regimen, 36 patients were entered. Thirty-one evaluable patients received a total of 200 and median of 6 (range: 2-8) cycles. Thirteen of the thirty-one had 27 DLEs within the first four cycles including grade 4 thrombocytopenia (n = 5), prolonged grade 4 neutropenia (n = 2), febrile neutropenia (n = 2), and omission of day 8 gemcitabine (n = 18). There was one patient death secondary to a wound abscess and febrile neutropenia. Myelosuppression as expected was the dose-limiting toxicity. CONCLUSION: The schedule of paclitaxel 135 mg/m(2) (day 1, 3 h), carboplatin AUC 5 (day 1), and gemcitabine 800 mg/m(2) (days 1 and 8) is feasible, with an acceptable toxicity profile. PMID- 14751145 TI - Fatty acid synthase is a marker of increased risk of recurrence in endometrial carcinoma. AB - PURPOSE: To explore the expression of fatty acid synthase (FAS) and human erythrocyte glucose transporter 1 (GLUT1) in endometrial carcinomas and to detect associations with clinicopathological features and prognosis. FAS and GLUT1 are two molecules involved in energy supply of normal cells. These markers are overexpressed in neoplastic tissues because of their increased necessity of energy. METHODS: Ninety-five patients with endometrial carcinoma were followed-up for an average period of 5 years. FAS and GLUT1 expressions were evaluated by immunohistochemistry on formalin-fixed paraffin-embedded tissues. Staining was determined with a semiquantitative method. Negative controls were obtained from patients submitted to hysterectomy for uterine prolapse. RESULTS: Eighty-five cases were endometrioid, 7 were serous, and 1 was a mucinous carcinoma. Seventy two cases (75%) were stage I, 12 (13%) were stage II, and 11 (12%) were stage III carcinomas. Sixteen (15%) carcinomas recurred. Nine patients (8%) died for cancer during the follow-up period. FAS expression was observed in 53 cases (56%). GLUT1 expression was observed in 32 (43%) cases. Statistical analysis revealed that FAS (P = 0.04) and stage (P = 0.001) of the disease were the only two independent predictors of recurrence. GLUT1 and other clinicopathologic parameters had no prognostic association. CONCLUSIONS: FAS is a reliable marker of clinically aggressive endometrial carcinomas. The knowledge of FAS expression in endometrial carcinomas is an important finding that may stratify patients into selected groups and determine therapeutic approaches for patient care. PMID- 14751146 TI - Lymphocyte composition of tumor draining lymph nodes from cervical and endometrial cancer patients. AB - OBJECTIVE: In the present study we compared the lymphocyte composition of tumor draining lymph nodes (TDLN) in 13 cervical (CC) and 26 endometrial cancer (EC) patients with special emphasis on the suppressive CD4(+) T cell subset constitutively expressing CD25 (T regulatory, Treg, cells). METHODS: Microscopically uninvolved TDLNs (n = 122) collected at different distances from the primary tumor site were used. Proximal TDLN included obturator, and internal and external iliac nodes. Distal TDLN included common iliac, presacral, and aortic nodes. Sixteen relevant lymphocyte subsets were assessed. The proliferative response to PHA and TCR crosslinking, the suppressive activity of Treg cells, and IFN-gamma and IL-4 production were also assessed. RESULTS: Compared to EC patients, TDLN from CC patients contained a higher proportion of naive CD4(+) and CD8(+) cells (P < 0.01), a lower proportion of CD4(+) pre effector cells (P < 0.05), and a higher proportion of the most potent Treg cell subset identified by coexpression of a high level of CD25 and CD152 (P < 0.05). Functionally, Treg cells were anergic and efficiently inhibited other T cell proliferation, thereby fulfilling the requirements of genuine Treg cells. The proliferative response of TDLN to both PHA and TCR crosslinking tended to be lower in CC than EC patients and was inversely related to the Treg cell content in both type of tumors. No differences were noticed between CC and EC patients in terms of IFN-gamma and IL-4 production. Using pairs of TDLN from individual patients, we showed that proximal TDLN in CC and EC patients displayed a higher CD4/CD8 ratio (P < 0.05 and 0.001, respectively) compared to distal TDLN of the same individual. By contrast, only in EC patients did proximal TDLN contained a proportion of Treg cells higher than distal TDLN (P < 0.02). CONCLUSIONS: The present data highlight differences in immune competency of TDLN in CC and EC patients, suggesting that the former is in a relative immunosuppressive status, and also underlines the importance of the proximity to the primary tumor site. PMID- 14751147 TI - Intratumoral effects of medroxy-progesterone on proliferation, apoptosis, and sex steroid receptors in endometrioid endometrial adenocarcinoma. AB - OBJECTIVE: The effects of progesterone on proliferation and apoptosis are studied in a scrutinized evaluation of endometrial carcinoma before, during, and after progesterone therapy. The heterogeneity of sex steroid expression as well as proliferation, indicated as Ki-67 index, is considered. METHODS: A total of 29 endometrial carcinomas were studied with in situ evaluation of Ki-67 proliferation marker, estrogen and progesterone receptors (ER and PR), and bcl-2 and p53 immunohistochemistry in the epithelial part of the tumor. In biopsy 1, before the therapy, Ki-67 ER, and PR were studied also in stroma. Apoptotic cells were morphologically identified in hematoxylin- and eosin-stained sections of the tumors and the apoptotic index (apoptotic cells per 1000 cells) was calculated. Chances in feature factors were mainly evaluated by repeated measures ANOVA. RESULTS: Proliferation (Ki-67) was decreased in grade 1 (G1) and grade 2 (G2) tumors during progesterone therapy both in overall evaluation (Ki) and particularly in the areas of maximal proliferation (Ki-max). No change was seen in G3 tumors. A decrease in PR expression in the areas of maximal expression for PR (PR-max) was also observed in G1 and G2 tumors. Apoptosis as well as bcl-2 and ER expression were unchanged during therapy and withdrawal. CONCLUSIONS: The effect of progesterone is seen only on proliferation in low-grade (G1 and G2) tumors. The coexistence of high PR expression in the foci of high proliferation may contribute to the effect in G1 and G2 tumors. No effect of progesterone is seen on apoptosis in tumors of any grade. PMID- 14751148 TI - An analysis of 84244 patients from the British Columbia cytology-colposcopy program. AB - OBJECTIVE: To determine the diagnostic correlation between referral cytology, initial biopsies and colposcopic impression in patients assessed in a provincial cytology screening program. METHODS: A retrospective review of the computerized cytology screening database for British Columbia (BC), to identify all patients having their first colposcopy between 1986 and 2000 in 24 participating clinics constituted the study population. 84244 patient records were identified for analysis. Colposcopies were performed mainly by 37 general gynecologists as part of a province-wide colposcopy program. Correlation of cytology, colposcopic impression and directed biopsies was performed. RESULTS: The colposcopic impression correlated with the referral cytology within one degree in over 90% of cases. Colposcopists felt cytology underestimated disease in 1.5% and overestimated disease in 8.3%. Cytology-histology correlation within one degree occurred in 82%. Cytology underestimated the result of the biopsies in 2.3% and appeared to overestimate disease in 16.1% of patients. Patients with HSIL cytology had corresponding lesions in 77%, with a further 4.9% having LSIL disease. The predictive accuracy of colposcopy increased with advancing severity of disease expected. As the degree of cytological abnormality worsened, the predictive accuracy of colposcopic diagnosis increased. CONCLUSIONS: Both cytology and colposcopy have high sensitivity but low to moderate specificity. Colposcopy is most accurate in identifying high-grade diseases. Colposcopic impression correlates closely with the cytology diagnosis and combining the two produces optimum results. PMID- 14751149 TI - Characterization of an anti-human ovarian carcinomaxanti-human CD3 bispecific single-chain antibody with an albumin-original interlinker. AB - OBJECTIVE: The objective of this study was to determine the properties of a single-chain bispecific antibody (scBsAb) against human ovarian carcinoma and to develop this agent for potential use in human ovarian cancer. METHODS: ELISA and FACS were performed to determine the antigen-binding properties of the scBsAb. Its abilities to retarget the pre-activated human peripheral blood mononuclear cells (PBMCs) to human ovarian carcinoma cell line SKOV3 cells and mediate their lysis in vitro were performed by a colorimetric MTT-based assay. Nude mice bearing human SKOV3 tumor xenografts were used to study the distribution and imaging of the scBsAb. Its pharmacokinetics in vivo was also studied in naive BALB/c mice. RESULTS: The scBsAb showed nearly identical ligand binding properties at each site relative to the individual monovalent single-chain antibody prototype molecules and could bridge SKOV3 and human T cell line Jurkat, which expresses CD3 antigens on the surface of cells together. It can also retarget the pre-activated PBMCs to SKOV3 cells and mediated their lysis in vitro effectively. Imaging and distribution study demonstrated that the antibody could target the tumor. Its elimination in vivo corresponded to second-order kinetics with a terminal half-life time (t(1/2)beta) of 7.7 h. CONCLUSION: This scBsAb with easy production and reasonable blood retention time should be developed for potential use in human ovarian cancer. PMID- 14751150 TI - Efficacy of intraperitoneal cisplatin as consolidation therapy in patients with pathologic complete remission following front-line therapy for epithelial ovarian cancer. Consolidative intraperitoneal cisplatin in ovarian cancer. AB - OBJECTIVE: The aim of this study is to evaluate the efficacy of intraperitoneal cisplatin as consolidation treatment in epithelian ovarian cancer patients with complete pathologic response following front-line platin-based chemotherapy. PATIENTS AND METHODS: Thirty patients who had no evidence of disease as assessed by second-look laparotomy following chemotherapy for stage III epithelial ovarian cancer were given three courses of intraperitoneal cisplatin (100 mg/m(2)) with three weekly intervals as consolidation therapy. RESULTS: Median age was 50 years. After a median follow-up period of 37 months, 16 patients are being followed with no evidence of disease. Eleven patients developed recurrent disease. Median disease-free survival was 50 months. Median overall survival is not reached. WHO grades 3-4 toxicity criteria were emesis in 19 patients (63.3%), abdominal pain in 5 (16.7%) and nephrotoxicity in 2 (6.7%) patients. Catheter related complications were infection/peritonitis in one and catheter malfunction in one patient. There were no serious hematologic side effects that required transfusions or caused treatment delays. None of the patients developed serious neurologic toxicity. Treatment had to be stopped early in four patients who refused further treatment due to abdominal pain, nausea ::and vomiting. Dose reductions were required in five patients. CONCLUSION: Our results suggest that intraperitoneal cisplatin is a feasible regimen that may provide a favorable outcome in terms of progression-free survival in patients with a complete pathologic response following front-line treatment for ovarian cancer. Further randomized trials are required to evaluate the role of consolidation treatment in this setting. PMID- 14751151 TI - Second-line chemotherapy with gemcitabine and carboplatin in paclitaxel pretreated, platinum-sensitive ovarian cancer patients. A Hellenic Cooperative Oncology Group Study. AB - OBJECTIVE: Patients with epithelial ovarian cancer (EOC) who relapse more than 6 months following completion of platinum-based primary chemotherapy are considered platinum-sensitive, and can be effectively retreated with cisplatin or carboplatin. The nucleoside analogue gemcitabine has proven activity in both platinum-sensitive and platinum-resistant disease. We conducted a phase II study using the combination of carboplatin and gemcitabine for the treatment of patients with relapsed platinum-sensitive and paclitaxel-pretreated EOC. METHODS: Forty-three patients were treated with gemcitabine 1000 mg/m(2), intravenously, over 30 min on days 1 and 8, and carboplatin at AUC 5 on day 1. Courses were administered every 3 weeks on an outpatient basis. RESULTS: Among 37 patients with measurable or evaluable disease, 15 (40.5%) achieved an objective response including 10 complete and 5 partial responses. The median overall survival was 24.5 months, and the median time to progression for all patients was 9 months. The treatment was well tolerated without toxic deaths; the most common toxicities were Grade 3 or 4 neutropenia, anemia, and thrombocytopenia that occurred in 69%, 26%, and 24% of patients, respectively. CONCLUSIONS: The combination of carboplatin and gemcitabine is a well-tolerated outpatient regimen with activity in patients with relapsed platinum-sensitive and paclitaxel-pretreated EOC. However, a randomized prospective study is justified to define whether the addition of gemcitabine to single-agent carboplatin results in improved efficacy in this subset of patients. PMID- 14751152 TI - In vitro chemoresistance and biomarker profiles are unique for histologic subtypes of epithelial ovarian cancer. AB - OBJECTIVES: To determine whether there is a relationship between histologic subtype of epithelial ovarian cancer and chemoresistance, we evaluated ovarian carcinomas of six histologic subtypes and correlated histology with in vitro drug response. Biomarker profiles (p53, Her-2 neu, and EGFR) were also evaluated to determine if their expression patterns were associated with histology. METHODS: In vitro drug response profiles for different histologic subsets of epithelial ovarian carcinomas exposed to standard relevant chemotherapy agents were determined in the Extreme Drug Resistance assay (EDR). Immunohistochemistry techniques were employed to determine biomarker expression. RESULTS: Of 5195 referred serial cases of epithelial ovarian cancer, there were 2660 papillary serous, 303 endometrioid, 142 mucinous, 102 clear cell, 952 undifferentiated carcinomas, and 42 tumors of low malignant potential. For the samples as a whole, the incidences of extreme drug resistance to the tested chemotherapeutic agents were cisplatin 10%, carboplatin 16%, cyclophosphamide 16%, doxorubicin 40%, gemcitabine 21%, paclitaxel 22%, and topotecan 13%. When compared to papillary serous tumors, mucinous tumors were more frequently resistant to cisplatin (10% vs. 18%) but less frequently resistant to topotecan (13% vs. 5%) and doxorubicin (42% vs. 16%). Endometrioid tumors were less resistant to cisplatin (10% vs. 6%) and doxorubicin (42% vs. 20%). Clear cell and undifferentiated tumors had the lowest rates of EDR to paclitaxel (13% and 18%) and cyclophosphamide (7% and 11%), while borderline tumors showed high rates of EDR to these agents (52% and 63%, respectively). With respect to biomarker profiles, mP53 was detected in 46%, Her-2 neu in 16%, and EGFR in 30% of the cases evaluated. As compared to all other subtypes, clear cell carcinomas had significantly higher Her-2 neu expression (19%). Relative to papillary serous carcinomas, borderline tumors exhibited significantly lower rates of mP53 expression (60% vs.17%). CONCLUSIONS: We found significant differences in the frequencies of extreme drug resistance to chemotherapeutic agents and biomarker expression among histologic subtypes of epithelial ovarian cancer. The data collected in this investigation may provide a guide for stratification of patients entering clinical trials based on histology and biomarker expression. PMID- 14751153 TI - Detection of CD8+ T cell responses to human papillomavirus type 16 antigens in women using imiquimod as a treatment for high-grade vulval intraepithelial neoplasia. AB - OBJECTIVES: To investigate CD8+ T cell reactivity to human papillomavirus (HPV) 16 antigens in patients with high-grade vulval intraepithelial neoplasia (VIN) before, during and after treatment with 5% imiquimod cream. METHODS: CD8-enriched responder cell populations were obtained from 10 patients with high-grade VIN using imiquimod cream as a treatment. Overlapping synthetic peptides covering the entire primary sequences of the HPV16 E6, E7 and E4 proteins were used to screen for CD8+ T cell responses using an ELISPOT assay of interferon (IFN)-gamma release. RESULTS: Reactivity to the proteins was detected in all patients on at least one occasion. With the exception of one patient, CD8+ T cell reactivity generally increased at some stage during treatment. The magnitude and specificities of responses changed over the treatment period. This was particularly noticeable in response to peptides derived from the E4 protein. CD8+ T cell reactivity to HPV16 E7 appeared to be dominant amongst women with high grade VIN. The magnitude and specificity of response had no correlation with clinical response to imiquimod. CONCLUSIONS: HPV16 specific CD8+ T cell activity was detected in patients with high-grade VIN. Imiquimod use appeared to increase the magnitude of the response and broaden the specificity of response in some patients. Despite the presence of these CD8+ T cells, the disease state persisted; therefore, a role for HPV-specific cytotoxic T cells (CTLs) in VIN resolution remains unproven. PMID- 14751154 TI - Ki-67 and vascular endothelial growth factor expression in uterine leiomyosarcoma. AB - OBJECTIVE: Uterine leiomyosarcomas (LMS) are rare female neoplasms of high malignant potential. The importance of different prognostic parameters in these malignancies remains controversial. The aim of the present study was to evaluate the prognostic value of Ki-67 and vascular endothelial growth factor (VEGF) in uterine LMS. METHODS: Ki-67 and VEGF expression was assessed using immunohistochemistry from paraffin-embedded tissue in 20 patients with uterine LMS. RESULTS: Ki-67 was expressed in 10 of 20 cases (50%) of uterine LMS. A statistically significant positive correlation was observed between vascular space involvement and Ki-67 expression (P = 0.05). There was no statistically significant relationship between Ki-67 expression and clinical stage (P = 0.38), recurrent disease (P = 0.86), and age (P = 1.0). An expression of VEGF could be observed in 1 of 20 (5%) cases. The median disease-free survival was 13 months (range: 3-23 months). Eleven (55%) patients died of the disease with a median overall survival of 47.4 months (range: 1-227 months), resulting in a 5-year overall survival rate of 45%. In a univariate analysis a statistically significant shortened disease-free survival in patients with Ki-67 positive tumors could be observed (P = 0.01). CONCLUSIONS: Our results suggest that the significant expression of Ki-67 in uterine LMS, correlating with vascular space involvement and being associated with a shortened disease-free survival, may be an indicator of the biological aggressiveness of these rare tumors. PMID- 14751155 TI - Activity of weekly paclitaxel in patients with advanced endometrial cancer previously treated with both a platinum agent and paclitaxel. AB - PURPOSE: The aim of this report is to describe the potential clinical utility of the weekly administration of paclitaxel in patients with endometrial cancer previously treated with a platinum agent and paclitaxel (delivered on an every 3 week schedule). METHODS: We briefly describe the clinical courses of three women with recurrent endometrial cancer who had prior exposure to platinum and paclitaxel, and who were subsequently treated with weekly paclitaxel in an effort to relieve significant cancer-related symptoms. RESULTS: In these individuals, the weekly administration of paclitaxel was reasonably well tolerated. There was evidence of both objective and subjective improvement in the status of the malignant process. CONCLUSION: The weekly administration of paclitaxel is a rational management approach in women with metastatic or recurrent endometrial cancer who have previously received treatment with both a platinum agent and paclitaxel. This delivery strategy should be further explored through the conduct of well-designed clinical trials, both in the primary and secondary chemotherapeutic management of this malignancy. PMID- 14751156 TI - Cytogenetic and molecular genetic characterization of immortalized human ovarian surface epithelial cell lines: consistent loss of chromosome 13 and amplification of chromosome 20. AB - OBJECTIVES: This study aimed at identifying the genetic events involved in immortalization of ovarian epithelial cells, which might be important steps in ovarian carcinogenesis. METHODS: The genetic profiles of five human ovarian surface epithelial (HOSE) cell lines immortalized by retroviral transfection of the human papillomavirus (HPV) E6/E7 genes were thoroughly characterized by chromosome banding and fluorescence in situ hybridization (FISH), at various passages pre- and post-crisis. RESULTS: In pre-crisis, most cells had simple, non clonal karyotypic changes. Telomere association was the commonest aberration, suggesting that tolermase dysfunction might be an important genetic event leading to cellular crisis. After immortalization post-crisis, however, the karyotypic patterns were non-random. Loss of genetic materials was a characteristic feature. The commonest numerical aberrations were -13, -14, -16, -17, -18, and +5. Among them, loss of chromosome 13 was common change observed in all lines. The only recurrent structural aberration was homogeneously staining regions (hsr) observed in three lines. FISH and combined binary ratio labeling (COBRA)-FISH showed in two cases that the hsrs were derived from chromosome 20. Clonal evolution was observed in four of the lines. In one line, hsr was the only change shared by all subclones, suggesting that it might be a primary event in cell immortalization. CONCLUSION: The results of the present study suggested that loss of chromosome 13 and the amplification of chromosome 20 might be early genetic events involved in ovarian cell immortalization, and might be useful targets for the study of genomic aberrations in ovarian carcinogenesis. PMID- 14751157 TI - Phase 2 trial of carboplatin, paclitaxel, and irinotecan in ovarian, fallopian tube, and primary peritoneal cancers. AB - OBJECTIVE: Our goal in this nonrandomized phase 2 trial was to evaluate the toxicity and obtain preliminary data on the potential efficacy of a novel three drug combination regimen (carboplatin-paclitaxel-irinotecan) when employed as initial therapy of advanced ovarian cancer or as second-line treatment in the setting of a prolonged (>or=12 months) treatment-free interval. METHODS: Patients with a histologically confirmed diagnosis of advanced ovarian cancer, primary adenocarcinoma of the peritoneum, or fallopian tube cancer were enrolled in the study. Patients received carboplatin (AUC 5), paclitaxel (150 mg/m(2) over 3 h), and irinotecan (100 mg/m(2) over 90 min). The three-drug combination was initially administered on an every 21-day schedule, but due to toxicity was subsequently changed to a 28-day program. RESULTS: A total of 30 patients were enrolled into this phase 2 trial. Twenty-three patients were chemotherapy naive, while 7 had received prior chemotherapy. Seventeen patients completed all six cycles of treatment. Eight patients (27%) were removed from the study after a median of three cycles due to toxicities. Seventeen patients (57%) experienced grade 4 neutropenia, with three individuals requiring hospitalization for neutropenic fever and dehydration. Grades 3 and 4 thrombocytopenia were experienced by three patients each. The principal nonhematologic toxicities were diarrhea (grade 3: three patients) and fatigue. The overall objective clinical response rate was 83%. CONCLUSIONS: The combination of carboplatin-paclitaxel irinotecan can be administered to women with advanced ovarian cancer with significant, but overall acceptable toxicity. Modification of the regimen from a 3-week to a 4-week schedule permits a greater percentage of patients to complete the program without experiencing excessive toxicity. The overall objective response rate observed in this trial is comparable to other combination regimens employed in this setting. Defining a place for this three-drug program in the standard management of ovarian cancer will require the conduct of an appropriately designed randomized trial. PMID- 14751158 TI - The tea polyphenol, (-)-epigallocatechin gallate effects on growth, apoptosis, and telomerase activity in cervical cell lines. AB - OBJECTIVE: To investigate the effect of the major tea polyphenol, (-) epigallocatechin gallate (EGCG) in cervical carcinogenesis. METHODS: Cell growth rate was examined after treatment for 4, 7, and 10 days with 0-100 microM EGCG in primary human endocervical cells (HEN), human papillomavirus type 18 (HPV 18) immortalized endocervical cell (HEN-18), ectocervical cell (HEC-18), serum adapted HEN-18 (HEN-18S), transformed HEC-18 (HEN-18T), and four cervical cancer cell lines. The effect of EGCG treatment was examined on dysplastic epithelium formation in organotypic culture, induction of apoptosis by DNA ladder assay and telomerase activity by PCR telomere extension assay. RESULTS: EGCG inhibited growth more than 90% in HEN-18 and HEC-18, whereas growth inhibition was less in ME180, TMCC-1, HeLa, SiHa, HEC-18T, and HEN-18S. In organotypic culture, thickness of epithelial multilayers was decreased in all EGCG-treated cells. EGCG resulted in apoptosis of HEN-18 or HEC-18, but not HEN-18S nor HEC-18T and inhibited telomerase activity in HEN-18 and HEC-18, as well as HEN-18S and HEC 18T. CONCLUSION: Our data suggest that EGCG prevents the carcinogenesis of cervical cancer, induces apoptosis and inhibited telomerase activity. The effect by EGCG treatment may be associated with the induction of apoptosis and telomerase inhibition in early cervical lesions. PMID- 14751159 TI - Characteristics of recurrence in patients who underwent lymphatic mapping for vulvar cancer. AB - OBJECTIVE: To evaluate patients with vulvar cancer who experienced a recurrence after undergoing lymphatic mapping and sentinel lymph node (SLN) biopsy. METHODS: We reviewed the records of 52 patients who underwent vulvectomy and lymphatic mapping with blue dye for treatment of vulvar cancer at our institution from 1993 to 1999 and identified patients who experienced recurrent disease. RESULTS: Fourteen (27%) of 52 patients experienced a recurrence. The patients' median age was 60 years (range 35-84 years). Nine patients had squamous lesions, four patients had melanoma, and one patient had Paget's disease with stromal invasion. Four tumors were stage T1, seven were T2, and three were T3. Eight lesions were located at the midline and six were lateral. Thirteen patients underwent superficial inguinal lymphadenectomy while one patient underwent SLN biopsy only. Postoperatively, seven patients underwent no further treatment, six underwent radiation therapy, and one patient underwent chemotherapy. The median follow-up was 46 months and the median disease-free interval was 21 months. Primary recurrence was in the vulva in eight patients (57%), in the groin in three patients (21%), and distant in three patients (21%). Nine of 32 (22%) squamous lesions recurred, four (57%) of seven melanomas recurred, and the sole patient with invasive Paget's recurred. Patient weight was found to be significantly different between patients who experienced a recurrence and those who did not (P = 0.05). At least one SLN was identified in 46 (88%) of the 52 patients. One (17%) of six patients in whom no SLN was identified experienced a recurrence, and 13 (28%) of 46 patients in whom a SLN was identified experienced a recurrence (P = 0.5). In the 41 patients with negative SLNs and negative non-SLNs, the recurrence rate was 24%; in the six patients with positive SLNs and negative non SLNs, the recurrence rate was 40%; and in the five patients with positive SLNs and positive non-SLNs, the recurrence rate was 40% (P = 0.6). No patients had a negative SLN and positive non-SLN. Of the three patients who experienced a recurrence in the groin, one had a negative SLN and negative non-SLN, one had a positive SLN and positive non-SLN, and one had no SLN identified and a negative non-SLN. CONCLUSIONS: . This heterogeneous group of patients who underwent lymphatic mapping with blue dye had similar patterns of recurrence to reported series of patients who did not undergo lymphatic mapping. Groin relapse following a negative SLN biopsy is of concern and suggests that long-term follow-up data are required before lymphatic mapping and SLN biopsy alone can be considered standard treatment for patients with vulvar cancer. PMID- 14751160 TI - The prognostic significance of thrombocytosis in epithelial ovarian carcinoma. AB - OBJECTIVE: The objective of this study was to determine the incidence of thrombocytosis in epithelial ovarian carcinoma and examine associations with clinico-pathologic features. Thrombocytosis (platelet counts >400 x 10(9)/l) has been identified as a poor prognostic factor in many cancers. Platelet-secreted factors may contribute to metastasis, invasion, and primary tumor growth. METHODS: One hundred eighty-three patients with invasive epithelial ovarian or primary peritoneal carcinomas were identified between January 1996 and December 2000. Records were retrospectively reviewed and data analyzed using chi(2), Student's t test, and Cox proportional hazards model; survival was analyzed by the method of Kaplan and Meier. RESULTS: Forty-one of 183 (22.4%) patients had thrombocytosis at primary diagnosis. Patients with preoperative thrombocytosis were found to have greater elevations of CA-125 (P = 0.026), more advanced stage disease (P = 0.016), higher grade tumors (P = 0.010), more frequent lymph node metastases (P = 0.018), and greater volume of ascites (P < 0.0001). One hundred sixty of 183 (87.4%) patients achieved optimal cytoreduction; patients with thrombocytosis demonstrated a greater likelihood of suboptimal resection (residual disease >1 cm; 19/41 vs. 4/142 in patients without thrombocytosis, P < 0.0001). Patients with thrombocytosis had a shorter disease-free interval (12 vs. 34 months, P < 0.0001) and overall survival (28 vs. 79 months, P < 0.0001). On multivariate analysis, thrombocytosis retained significance as a poor prognostic indicator in patients with stage III and IVA disease (P = 0.04). CONCLUSIONS: Thrombocytosis is a frequent preoperative finding in ovarian and peritoneal carcinomas and may be a marker of aggressive tumor biology. PMID- 14751161 TI - The role of radical parametrectomy in the treatment of occult cervical carcinoma after extrafascial hysterectomy. AB - OBJECTIVES: To assess the morbidity and efficacy of radical parametrectomy (RP) performed following extrafascial hysterectomy in patients with occult cervical carcinoma. METHODS: An IRB approved retrospective chart review identified 23 patients that underwent RP with pelvic and/or para-aortic lymphadenectomy and upper vaginectomy. Data were collected on demographics, tumor stage, grade, histology, indication for hysterectomy, surgical findings, complications, recurrence, and survival. RESULTS: Of the 23 patients, 2 patients had a stage IA(2) lesion while 21 patients had a stage IB(1) lesion. There were 5 patients with a grade 1 tumor, 10 with grade 2, 4 with grade 3, and 4 with unknown grade. Median age was 41 years (range 27-59). The most common indication (48%) for extrafascial hysterectomy was CIS of the cervix. Four patients (17%) had metastasis to pelvic nodes or evidence of tumor at the margin at the time of RP. Three of these 4 patients with a positive specimen received adjuvant radiation and all are alive (mean follow-up 66 months). One patient declined radiation and is alive at 42 months. There were 7 (30%) operative complications: Most notably 4 patients received blood transfusions. Two of 19 patients (11%) with no residual tumor in RP specimen recurred and 1 patient was salvaged with radiation (follow up 103 months). With a median follow-up of 61 months (range 9-103), overall 5 year survival is 96%. CONCLUSIONS: RP is an acceptable option for patients diagnosed with an occult cervical carcinoma at the time of extrafascial hysterectomy. Careful selection of RP for patients unlikely to have residual tumor will obviate the need for radiation in most instances. PMID- 14751162 TI - The gastrointestinal complications of the Miami Pouch: a review of 77 cases. AB - OBJECTIVES: To describe the gastrointestinal (GI) complications associated with the Miami Pouch (MP), a continent ileocolonic urinary reservoir. METHODS: A retrospective chart review of patients who underwent a MP from 1988 to 1997 at the University of Miami, School of Medicine, was employed. Data was analyzed in terms of early and late (beyond 6 weeks) GI complications resulting directly from the operation. RESULTS: Seventy-seven patients underwent a MP, a form of continent urinary diversion. Seventy-two patients (93.5%) were previously radiated. The perioperative mortality rate was 11.7%. Twenty (26%) patients developed a GI complication (17 late and 3 early), and 5 (6.5%) were directly as a result of the MP. Twelve recto-vaginal and 1 recto-neo-vaginal fistulas were identified. All but one was considered as late. Three (3.9%) patients developed colo-MP fistulas (3, 5, and 14 months). All three patients failed conservative management and required reoperation. Two patients developed enterocutaneous fistulas (3 and 5 months). One patient developed breakdown of the ileotransverse colon anastomosis on postoperative day 12 and required reoperation with bowel resection and an ileostomy. She expired from intraabdominal sepsis. Finally, 1 patient developed short bowel syndrome secondary to an expanding hematoma in the small bowel mesentery. CONCLUSIONS: . The GI complication rate attributed directly to the MP is low (6.5%). Prompt recognition is the key to successful management of these complications. The majority of these complications are considered as late and do not occur in the immediate postoperative period. Conservative management of GI-MP fistulas is not successful and necessitates reoperation. PMID- 14751163 TI - Presence of multiple human papillomavirus types in cervical samples from HIV infected women. AB - OBJECTIVES: The aim of this study was to detect and identify human papillomavirus (HPV) genotypes on a population of women infected by the human immunodeficiency virus (HIV) and to investigate the role of multiple infections on cervical dysplasia. METHODS: Two hundred and fifty-five HIV-infected women were enrolled on a study to evaluate the prevalence of HPV and cervical intra-epithelial neoplasia (CIN). A group of HIV-negative women with confirmed CIN diagnosis was included for comparison. A polymerase chain reaction (PCR)-reverse hybridization method was applied to detect and precisely identify HPV types, specifically multiple infections. RESULTS: On HIV patients, an altered Pap smear confirmed by biopsy was observed on 45 (18%); HPV-DNA prevalence was 87% (223/255), with 45% (116/255) infected by more than two types. In contrast, HPV-DNA was detected in all 36 women of the control group but only 3 were infected by more than two types. Cervical dysplasia was associated with low CD4 counts and elevated high risk HPV viral load. However, the presence of multiple HPV types did not correlate with the degree of immune suppression or the presence of cervical lesions. CONCLUSIONS: Infection with multiple HPV types is a rather frequent finding on Brazilian HIV-infected women. On this population, concomitant infection with three or more HPV types does not seem to confer an additional risk of cervical dysplasia in comparison to single/double infections, nor to be related to more severe immunesuppresion. PMID- 14751164 TI - Symptoms and diagnosis of borderline, early and advanced epithelial ovarian cancer. AB - OBJECTIVE: To document symptoms associated with borderline, early and advanced ovarian cancer and identify personal characteristics associated with early versus late diagnosis. METHODS: Information concerning symptoms and diagnosis history was available from 811 women with ovarian cancer who took part in an Australian case-control study in the early 1990s. Women were classified into three groups for comparison based on their diagnosis: borderline, early (stage I-II) and advanced (stage III-IV) invasive cancer. RESULTS: Sixteen percent of women with borderline tumors, 7% with early cancer and 4% with advanced cancer experienced no symptoms before diagnosis (P < 0.0001). Among women with symptoms, abdominal pain (44%) or swelling (39%) were most frequently reported; an abdominal mass (12%) and gynecological symptoms (12%) were less common. Compared to advanced stage cancer, women with early stage cancer were more likely to report an abdominal mass or urinary symptoms but less likely to report gastrointestinal problems or general malaise. General malaise and 'other' symptoms were least common in borderline disease. Older women, and those with higher parity or a family history of breast or ovarian cancer, were more likely to be diagnosed at an advanced stage of disease. CONCLUSIONS: Women who experience persistent or recurrent abdominal symptoms, particularly swelling and/or pain should be encouraged to seek medical attention and physicians should be alert to the possibility of ovarian cancer even in the absence of an abdominal mass. Further information about the prevalence of these symptoms in the general population is essential to assist physicians in patient management. PMID- 14751165 TI - Optimal management for surgically Stage 1 serous cancer of the uterus. AB - OBJECTIVE: To describe the outcomes of patients who have undergone well-conducted surgery and found to have Stage 1 serous uterine cancer. METHODS: This retrospective cohort study includes women who have been treated for Stage 1 serous cancer of the uterus from 1985 to 2001. Cases were included from the regional cancer centers in Hamilton, London, Sunnybrook Toronto and Cancer Care Manitoba. RESULTS: Forty-three women met the inclusion criteria: Complete surgical staging (n = 27), surgery followed by pelvic radiation therapy (n = 4), surgery followed by whole abdominal radiation therapy (n = 6), surgery followed by adjuvant chemotherapy (n = 6). Patient age or depth of invasion did not influence survival. Progression free interval was 22 months (SD = 14.29). Recurrence rate was highest for adjuvant chemotherapy (66%). Survival was assessed by treatment modality and a statistically significant poorer survival was seen in the adjuvant chemotherapy group (OR 17.5; 95% CI 1.3-227.6). No comment can be made on a superior treatment regimen given the small numbers in each treatment strata. CONCLUSION: This study supports the findings of others in the literature. In a group of patients where surgical staging shows limited disease (i.e., surgically Stage 1 disease), then surgery alone appears to be adequate treatment. PMID- 14751167 TI - Epidemiology of choriocarcinoma in Finland, 1953 to 1999. AB - OBJECTIVE: Choriocarcinoma is a rare tumor with remarkable differences in the incidence in various parts of the world. The available data do not elucidate recent time trends in the incidence of the disease in Western countries. We studied the epidemiology of choriocarcinoma in Finland over a period of 47 years. METHODS: Incidence rates for choriocarcinoma from 1953 to 1999, according to the population-based Finnish Cancer Registry, were calculated per number of deliveries, obtained from the National Research and Development Center for Welfare and Health. Standardized incidence ratios (SIRs) for the years 1971 through 1995 for women born between 1906 and 1945 were calculated by occupation taken from the 1970 Population Census. RESULTS: The incidence of choriocarcinoma was 40/10(6) deliveries for the study period 1953-1999 as a whole. The respective incidence rate calculated per female population and adjusted for age to the world standard population was 1.3/10(6). The incidence, per number of deliveries, was remarkably higher in women above 40 years as compared to younger women. There was a decline in the incidence of the disease in women between 25 and 39 years of age from 53/10(6) deliveries in 1953-1984 to 26/10(6) deliveries in 1985-1999. High occupation-specific risks were observed for nurses (SIR 7.8; 95% confidence interval 2.1-20) and agricultural workers (SIR 11; 95% confidence interval 1.4 40). CONCLUSION: The incidence of choriocarcinoma in Finland is similar to that reported earlier for other Western countries. The recent decline in the incidence of the disease, the enormous increase in the risk among old fertile women, and clustering of cases to certain occupations should be targets of future studies. PMID- 14751166 TI - Cell cycle and apoptotic proteins in relation to ovarian epithelial morphology. AB - OBJECTIVE: To assess the relationship between the expression of cell cycle and apoptotic proteins and the morphological appearance of the surface epithelium in non-neoplastic ovaries. METHODS: The subjects for this study were 79 women who had undergone oophorectomy for benign conditions at the North Middlesex Hospital, London, and Royal Free Hospital, London, and whose ovaries had been reported on routine histology as entirely normal or containing physiological cysts or endometriosis. The epithelial morphology was reassessed on haematoxylin and eosin stained paraffin wax sections using nine cytological and architectural parameters associated with premalignant intraepithelial changes. A 'score' was obtained for each ovary. Expression of p53, Ki67, cyclin D1 and Bcl-2 in the surface, cystic and endometriotic epithelium was assessed in corresponding sections using standard immunohistochemistry. RESULTS: The median score for the morphological changes was significantly higher in the sections, which expressed p53 compared to those which did not. This difference remained significant in a subanalysis of the sections, which did not contain endometriosis. No relationship was identified between the morphological score and the expression of Ki67, Bcl-2 and cyclin D1. CONCLUSION: Increased intraepithelial abnormality as assessed by an epithelial morphological score of ovarian sections is associated with expression of the p53 cell cycle protein. This lends credence to the hypothesis that the ovarian surface or cystic epithelium goes through an identifiable precursor or "premalignant" phase before the development of invasive disease. Further work is required to characterise the changes that take place before the development of malignancy in ovarian epithelium. PMID- 14751168 TI - Detection of disseminated tumor cells in peripheral blood of patients with breast cancer: correlation to nodal status and occurrence of metastases. AB - OBJECTIVE: Soon after breast cancer becomes invasive, it sheds cancer cells into the blood stream or the cancer cells are spread via lymphatic vessels. The early and unambiguous detection of these disseminated tumor cells (DTC) is of importance for the evaluation of the tumor process and for monitoring therapy response. The detection of disseminated tumor cells by immunocytochemistry (ICC) without previously performing tumor cell enrichment is time consuming and may miss a considerable part of these cells. Therefore, we have applied a negative immunomagnetic enrichment of disseminated tumor cells from the peripheral blood of patients with breast cancer by the depletion of CD45(+) leukocytes using automated magnetic activated cell separation (autoMACS). METHODS: One hundred twenty-five blood samples from 83 breast cancer patients were investigated for occurrence of disseminated tumor cells by autoMACS technique and immunocytochemical cytokeratin staining. Frequency of disseminated tumor cells was analyzed statistically for correlation to clinical data. RESULTS: Thirty three of the 125 blood samples (26%) originating from 29 of 83 breast cancer patients (35%) carried cytokeratin positive (CK(+)) tumor cells. The occurrence of CK(+) tumor cells correlated significantly with the nodal status (P = 0.009) and with the occurrence of metastases at the time of primary tumor resection (P = 0.003). CONCLUSIONS: The finding that occurrence DTC detected in peripheral blood of breast cancer patients correlated with nodal stage and metastases is described for the first time. It suggests that disseminated tumor cells identified in peripheral blood by autoMACS are associated with tumor characteristics of breast cancer. PMID- 14751169 TI - Prognostic factors in adenocarcinoma of the uterine cervix. AB - OBJECTIVE: To determine the behaviour of adenocarcinomas of the uterine cervix during the last 10 years in the southwest region of the Netherlands, and to determine prognostic factors. METHODS: Three hundred and five cases of primary cervical adenocarcinomas (ACs) diagnosed between 1989 and 1999 in the region of Rotterdam, The Netherlands, were retrieved. Clinical and pathological data were reviewed and analysed. RESULTS: Mean age at presentation was 52 years. The mean follow-up time for surviving patients was 61 months. The overall survival was 60% at 5 years. The 5-year survival rates for stages I and II were, respectively, 79% and 37%. The 5-year survival rates for stages III and IV were less than 9%. Using univariate analysis stage, grade, age < 35 years and histological type were significant prognostic factors. In the group of patients who underwent surgery (n = 200), stages I-IIA, lymph node metastases, lymph-vascular-space-invasion (LVSI) and depth of stromal invasion were significant for survival. For patients with stages I and II-A disease, survival was significantly better where the primary treatment was surgical as opposed to primary radiotherapy (P = 0.002). Using multivariate analysis, only stage, grade and lymph node metastases remained significant independent predictors for survival. CONCLUSIONS: This report about cervical adenocarcinoma in the southwest region of The Netherlands shows similar results for survival to previous reports. Longest survival was for patients with early stage disease, younger patients and after primary surgery. We found FIGO stage, grade and lymph node metastases of significant prognostic value for survival in cervical adenocarcinoma. PMID- 14751170 TI - Expression of cellular apoptosis susceptibility protein in serous ovarian carcinoma: a clinicopathologic and immunohistochemical study. AB - OBJECTIVE: This study investigated the expression of cellular apoptosis susceptibility protein (CAS) in serous ovarian carcinoma by immunohistochemistry and compared it with topoisomerase IIalpha (topo IIalpha), bcl-2, bcl-x, the frequency of apoptotic bodies (ABI), mitotic activity, and c-erbB-2 with regard to clinicopathologic variables. METHODS: Formalin-fixed, paraffin-embedded archival tissue sections of 10 benign serous cystadenomas, 10 serous neoplasms of low malignant potential (LMP), and 41 serous ovarian carcinomas were immunostained with antibodies to CAS, bcl-x, topo IIalpha, bcl-2, and c-erbB-2. Immunostaining for CAS, bcl-x, and bcl-2 was scored concerning approximate percentage of positive tumor cells and relative staining intensity. For c-erbB-2, at least 10% of tumor cells had to display membrane staining. Topo IIalpha labeling indices were quantitated as the percentage of positively stained nuclei in 1000 tumor cells. ABIs were reported as the number of apoptotic bodies in 1000 tumor cells, mitotic activity as mitotic figures per 10 high power fields. RESULTS: CAS expression was negative in serous cystadenomas and LMP. In contrast, moderate or strong immunostaining was observed in 34 of 41 cases (83%) of serous carcinomas. CAS immunoreactivity was positively related with ABI (P = 0.0170), mitotic activity (P < 0.0001), c-erbB-2 (P = 0.0153), grade (P = 0.0107), and adverse outcome (P = 0.0035), but not with FIGO stage, topo IIalpha, bcl-2, and bcl-x. Other variables indicating outcome were topo IIalpha, c-erbB-2, and ABI. CONCLUSIONS: CAS is frequently upregulated in serous ovarian carcinomas, correlated with apoptosis and mitotic activity, and relevant prognostically. CAS protein expression may serve as a marker of aggressive tumor behavior in serous ovarian carcinomas. PMID- 14751171 TI - Comparative study of laparoscopico-vaginal radical hysterectomy and abdominal radical hysterectomy in patients with early cervical cancer. AB - OBJECTIVE: The objective of this study was to compare the outcomes of patients treated by laparoscopico-vaginal (modified) radical hysterectomy (LVMRH) to those of patients treated by abdominal radical hysterectomy (RH). METHODS: From 1997 to 2002, we performed 37 cases of LVMRH + pelvic lymph node dissection (PLND) and 47 cases of laparoscopico-vaginal radical hysterectomy (LVRH) with paraaortic lymph node sampling + PLND. Inclusion criteria for the laparoscopic surgery were patients with FIGO stage IA1 to IB1, for exocervical mass of grossly less than 2 cm. As a control, we selected 46 cases for MRH group and 96 cases for RH group. RESULTS: Operating time, the number of lymph nodes obtained and the rate of complications were similar in both groups. The hospital stay was significantly shorter in laparoscopic group. Four (8.5%) of 47 LVRH patients and 2 (2.1%) of 96 RH patients had recurrences. Recurrence-free survival in RH group was significantly higher than LVRH group (P = 0.0194). In LVRH group, patients with large tumor volume (>or=4.2 cm(3)) had significantly higher recurrence rate of 42.9% (3/7) than those with small volume (1/40) (P = 0.0021). The 3-year progression-free survivals were 97.1% in LVRH group (<4.2 cm(3)) and 98.9% in RH group. CONCLUSION: Laparoscopic surgery for the treatment of early cervical cancer is a safe and effective alternative to conventional RH. Considering the higher recurrence rate in patients with large tumor volume, it would be better if laparoscopic surgery is limited to patients with small volume disease (tumor diameter <2 cm or volume <4.2 cm(3)). PMID- 14751172 TI - Correlation between responsiveness of neoadjuvant chemotherapy and apoptosis associated proteins for cervical adenocarcinoma. AB - OBJECTIVE: Adenocarcinoma of the uterine cervix appears to be increasing in prevalence and it has been suggested that these tumors tend to be less sensitive to radiation therapy and to chemotherapy than squamous carcinomas. In the present study, 29 patients with locally advanced cervical adenocarcinoma (bulky IB-IVB) were treated with neoadjuvant chemotherapy (NAC) using cisplatin, aclacinomycin-A and mitomycin-C, followed by radical surgery or irradiation. METHODS: To predict the prognosis and response to the chemotherapy, the expression of apoptosis associated-proteins, p53, p21WAF1/CIP1, Bcl-2 and activated caspase-3 was evaluated for tumor samples by immunohistochemistry. RESULTS: Of the analyzed clinicopathological factors, the overexpression of p53 was frequently observed in endocervical-type adenocarcinoma, nonresponders to chemotherapy and the grade 0 histologic effect of the chemotherapy. Positive staining of Bcl-2 was frequently observed in the early stage and had a better prognosis than for patients with the negative staining; however, there was no correlation between responders and nonresponders to chemotherapy. The expression of p21WAF1/CIP1 and caspase-3 was not correlated to the clinicopathological factors. CONCLUSION: In this study, the overexpression of p53 was found to be a factor to predict the chemoresistance and positive expression of Bcl-2 indicated as a better prognostic value. For p21WAF1/CIP1 and caspase-3, further analysis is necessary. PMID- 14751173 TI - Patterns and outcomes of chemotherapy for elderly patients with stage II ovarian cancer: a population-based study. AB - BACKGROUND: The optimal treatment for patients with stage II ovarian cancer is controversial, although most experts recommend adjuvant chemotherapy. The purpose of this study was to assess the patterns of use of chemotherapy in women with stage II ovarian cancer, and to compare the survival of treated and untreated patients aged 65+ years in a population-based sample. METHODS: Using the Surveillance, Epidemiology, and End Results (SEER)-Medicare database of cancers diagnosed in approximately 14% of the U.S. population, we identified women who were diagnosed with stage II ovarian cancer between 1992 and 1996, survived >or=120 days beyond diagnosis, and were >or=65 years of age. Multivariate regression was used to compare those treated to those not treated with chemotherapy. Cox proportional hazards regression models were used to analyze patient survival. RESULTS: Of 236 women with stage II ovarian cancer, 160 (67.8%) received chemotherapy, and 118 (50%) received platinum-based regimens. Younger patients and those with higher-grade tumors were more likely to receive chemotherapy. The adjusted hazards ratio for mortality associated with any chemotherapy use was 0.67 (95% CI, 0.45-0.98), corresponding to an increase in median survival from 28 months to 35 months (P < 0.0001). CONCLUSIONS: This observational study found that most patients aged >or=65 years and diagnosed with stage II ovarian cancer between 1992 and 1996 were treated with chemotherapy. Grade and younger age were the most significant predictors of treatment, and treatment was associated with a 5-year mortality reduction of 33%. These findings are not definitive, but they may provide some guidance in the absence of randomized trials of adjuvant chemotherapy for older women with stage II ovarian cancer. PMID- 14751174 TI - Screening for HIV infection in women with newly diagnosed cervical cancer. AB - OBJECTIVE: This study aimed to investigate the prevalence of human immunodeficiency virus (HIV) infection in women with cervical cancer and to decide whether HIV testing should be included as part of the initial work-up of invasive cervical cancer. METHODS: This was a prospective case controlled study. Women with newly diagnosed cervical cancer were tested and age-matched women with benign gynecologic conditions were selected as control. Serum samples were tested for HIV I and HIV II antibodies at diagnosis and 3 months later. RESULTS: Two hundred twenty-five consecutive patients with newly diagnosed cervical cancer and two hundred twenty-three age-matched women with benign gynaecological conditions were tested. In comparison with the control, women with cervical cancer started sexual intercourse at an earlier age and had a statistically significant higher number of sexual partners. All the HIV tests were negative. The HIV prevalence rate was estimated to be 0.0-1.6% for both groups. CONCLUSION: HIV screening should not be a part of the initial work-up for cervical cancer in a population with a low background prevalence. PMID- 14751175 TI - Expression of estrogen receptor coregulators in normal and malignant human endometrium. AB - OBJECTIVES: To compare the expression of nuclear receptor coregulators in normal and malignant human endometrium and to identify any relationship to grade, stage, age, depth of myometrial invasion, estrogen receptor alpha (ERalpha), or progesterone receptor (PR) expression. METHODS: Gene expression of SRC-1, SRC-2, SRC-3, N-CoR, SMRT, ERalpha, and PR was measured in 26 samples of normal endometrium and 30 primary endometrial carcinomas using real-time RT-PCR. ERalpha protein expression of each tissue was also measured by Western blot. RESULTS: . All coregulators showed significantly increased mRNA expression in endometrial carcinoma as compared to normal endometrium. The mRNA expression of each coregulator showed a high correlation with ERalpha mRNA, PR mRNA, and with the other coregulators in both normal and malignant endometrium. In the normal endometrium, SRC-1 mRNA expression was positively correlated with ERalpha protein expression and SRC-3 mRNA expression was positively correlated with patient age. No relationship was found between coregulator mRNA expression and grade, stage, or depth of myometrial invasion. CONCLUSION: The nuclear receptor coregulators SRC-1, SRC-2, SRC-3, N-CoR, and SMRT were found to be up-regulated in malignant endometrium. Our findings suggest that these proteins may have a role in the development of endometrial carcinoma. PMID- 14751176 TI - An in vitro study of the inhibitory activity of gemcitabine and platinum agents in human endometrial carcinoma cell lines. AB - OBJECTIVE: To determine the cytotoxic activity of gemcitabine on a panel of endometrial cancer cell lines and to determine if the addition of platinum agents enhanced this activity. METHODS: A panel of four endometrial carcinoma cell lines was treated with variable concentrations of gemcitabine, gemcitabine and cisplatin, or gemcitabine and carboplatin. 3-(4,5-Cimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide (MTT) assays were performed to determine the IC(50). RESULTS: Each cell line evaluated revealed a dose-dependent decrease in cell growth for gemcitabine alone and in combination with platinum agents. The IC(50) of gemcitabine with cisplatin was significantly lower than that of gemcitabine alone for all cell lines, including the platinum-resistant line. CONCLUSION: Gemcitabine exerts in vitro cytotoxicity on endometrial cancer cell lines, and the combination of gemcitabine and cisplatin is superior to gemcitabine alone. This may represent a promising option for treatment of endometrial cancer. PMID- 14751177 TI - Reproductive and dietary risk factors for epithelial ovarian cancer in China. AB - OBJECTIVE: A case-control study was conducted to investigate the effects of reproductive and dietary risk factors on ovarian cancer risk in China. METHODS: Cases were 254 patients with histologically confirmed epithelial ovarian cancer. Controls were 652 women without neoplasm and long-term dietary modifications. Information was collected using a structured questionnaire on sociodemographic and reproductive characteristics, diet, medical and family cancer history. The risks of ovarian cancer were assessed by multivariate logistic regression analysis. RESULTS: The adjusted odds ratios (OR) for women having at least two full-term pregnancies, two or more incomplete pregnancies, and first full-term pregnancy at 21-25 years of age were 0.45 (95% CI 0.3-0.8), 0.56 (95% CI 0.4 0.8), and 0.40 (95% CI 0.2-0.8), respectively, compared with nulliparity. The OR of ever lactation was 0.50 (95% CI 0.3-0.8) and oral contraceptive was 0.48 (95% CI 0.3-0.7), while postmenopausal women appeared to have an increased risk with OR 1.48 (95% CI 1.0-2.3). For the highest versus the lowest quartile intakes of nutrients, the OR were 2.17 (95% CI 1.3-3.8) for fat, 0.36 (95% CI 0.2-0.6) for fibre, 0.26 (95% CI 0.2-0.5) for carotene, 1.59 (95% CI 0.9-2.7) for retinol, 0.31 (95% CI 0.2-0.5) for vitamin C, and 0.41 (95% CI 0.2-0.7) for vitamin E, with significant dose-response relationships. CONCLUSION: It is evident that full term and incomplete pregnancies, lactation, and oral contraceptive use can reduce the ovarian cancer risk. Moreover, consumption of foods low in fat but high in fibre, carotene and vitamins appears to be protective against ovarian cancer in Chinese women. PMID- 14751178 TI - Role of laparoscopy in the evaluation of the adnexa in patients with stage IV breast cancer. AB - OBJECTIVE: To describe the utility of laparoscopic evaluation of adnexal masses in women with stage IV breast cancer. METHODS: A retrospective review of gynecologic and breast surgery databases at our institution was performed to identify patients with stage IV breast cancer who underwent surgical evaluation for an adnexal mass or bilateral salpingo-oophorectomy (BSO) between January 1986 and August 2002. Patient demographics and operative and pathologic findings were reviewed. RESULTS: Thirty-one patients were identified. Median age was 47 years (range, 25-79 years). Pathology of the primary breast tumor was infiltrating ductal carcinoma in 58%, invasive lobular carcinoma in 29%, and unspecified in 13%. Median time from diagnosis of stage IV breast cancer to surgical evaluation of the adnexa was 15 months (range, 0-106 months). Surgery consisted of planned laparotomy in four patients and laparoscopic evaluation in 27 patients. Six patients had laparoscopic BSO for hormonal ablation. The remaining 21 patients had laparoscopic evaluation of an adnexal mass. Conversion to laparotomy occurred in three patients based on intraoperative findings of suspected primary ovarian cancer and for technical reasons in one patient. Overall, metastatic breast cancer was diagnosed in 21 (68%) of 31 patients, including two patients with occult metastases undergoing BSO for hormonal ablation. Primary ovarian cancer was found in 3 (10%) of 31 patients, and 7 (22%) of 31 patients had benign findings. Pathologic intraoperative frozen section was obtained in 21 (84%) of 25 patients undergoing laparoscopic evaluation for an adnexal mass. Intraoperative frozen section was concordant with final pathology in 20 (95%) of 21 patients (18 on laparoscopic evaluation, two on laparotomy). CONCLUSIONS: The majority of patients with stage IV breast cancer who present with an adnexal mass will be diagnosed with metastatic breast cancer. A small subset of patients will be diagnosed with primary ovarian cancer; thus, the evaluation of an adnexal mass even in this stage IV setting is warranted. Accurate diagnosis of metastatic breast cancer versus ovarian cancer can be made laparoscopically, thereby avoiding laparotomy in the metastatic breast cancer setting. PMID- 14751180 TI - Multicystic benign mesothelioma of the peritoneum presenting as postmenopausal bleeding and a solitary pelvic cyst--a case report. AB - BACKGROUND: All non-invasive diagnostic algorithms suffer from diagnostic uncertainty in discriminating benign from malignant adnexal masses. CASE: We present a case where the identification of a large solitary multiloculated pelvic cyst in a woman with postmenopausal bleeding was shown to be multifocal multicystic benign mesothelioma (MBM) of the peritoneum at laparotomy. CONCLUSION: We wish to alert gynaecologists of this rare association and the unique appearance of mesothelioma lesions, which many would not instantly recognise. There is a lack of consensus on the treatment and follow-up of MBM. Most authorities consider curative treatment of MBM is achievable through complete disease cytoreduction (peritonectomy) as carried out in the case. PMID- 14751179 TI - Laparoscopic enterolysis before adjuvant radiotherapy in a case of endometrial cancer. AB - CASE: A 33-year-old woman presenting with endometrial cancer stage FIGO Ic. After hysterectomy, bilateral oophorectomy and lymph node dissection, combined radiotherapy was indicated. Computer tomography scan was performed to plan radiotherapy and large bowel adhesions were found. To prevent bowel complications during radiation therapy, laparoscopic enterolysis of the adhesions was performed. We were able to start and apply radiation therapy only 22 days after initial laparotomy. Our patient had a history of deep vein thrombosis as a result of oral contraceptive medication. The endoscopic treatment enabled us to shorten the time of immobilization and bed rest substantially. CONCLUSION: Laparoscopy is an appropriate method to prevent complications due to radiotherapy. PMID- 14751181 TI - An unusual admixture of neoplastic and metaplastic lesions of the female genital tract in the Peutz-Jeghers Syndrome. AB - BACKGROUND: Peutz-Jeghers Syndrome (PJS) is a rare autosomal dominant condition with variable penetrance characterized by gastrointestinal hamartomatous polyps and mucocutaneous pigmentation. Patients with PJS have an increased risk for breast, gastrointestinal and female genital tract cancers. CASE: Multiple genital tract neoplasms in a 41-year-old Italian woman with PJS are described. The patient presented with abdominal pain due to intussusception. A CT scan of the abdomen also showed a left adnexal mass, diagnosed as ovarian mixed serous and mucinous borderline tumor. An ovarian microscopic sex cord tumor with annular tubules (SCTAT) was incidentally diagnosed together with a minimal deviation mucinous adenocarcinoma of the uterine cervix. Also areas of typical hyperplasia of the tubal mucosa with mucinous metaplasia were found. CONCLUSION: This appears to be one of the rare cases reported in literature in which PJS is complicated by multiple and contemporaneous genital tract tumors and rare histological findings. The clinical significance of recurrence of these unusual genital tract tumors and histological alterations in PJS patients is reviewed. PMID- 14751182 TI - An unusual cause of adnexal mass: fallopian tube schwannoma. AB - BACKGROUND: Schwannoma is a well-defined, usually benign tumor arising from the Schwann cells in the nerve sheath. It can present in any location as a solitary mass; however, it is rarely found in the pelvis. CASE: We report a 40-year-old female patient with a pelvic mass, which revealed tubal schwannoma after complete resection of the mass and uterus by laparotomy. CONCLUSION: Schwannomas are thought to result from a proliferation of perineural cells. Although origin along the nerves of the retroperitoneal space is not uncommon, these tumors rarely present as pelvic masses. PMID- 14751183 TI - Conservative management of a uterine tumor resembling an ovarian sex cord-stromal tumor. AB - BACKGROUND: Sex cord-stromal tumors usually arise in the ovary but may arise in the uterus. Only 55 cases have been reported thus far. A hysterectomy is typically recommended. We report a case of uterine tumor resembling an ovarian sex cord-stromal tumor managed conservatively. CASE: A 32-year-old nulliparous woman presented with infertility secondary to suspected uterine leiomyomata. A laparoscopic resection of the presumed leiomyomata was performed, and the pathology showed a uterine tumor resembling an ovarian sex cord tumor. Because of the patient's desire to preserve fertility, a hysterectomy was not performed. She has been followed for the past 18 months, and there is no evidence of tumor recurrence. CONCLUSION: Resection of the tumor without hysterectomy and close follow-up may preserve fertility without compromising survival in patients with uterine tumors resembling ovarian sex cord-stromal tumors. PMID- 14751184 TI - Gliomatosis peritonei and teratomatous implant with carcinomatous transformation presenting 54 years following oophorectomy for dermoid cyst. AB - BACKGROUND: Gliomatosis peritonei is a rare condition associated with ovarian teratomas in which benign glial implants are identified on the peritoneal surfaces of the abdomen. The implants have been identified at initial surgery and at second-look laparotomy. CASE: Here, we present a case of an 82-year-old female who was diagnosed with gliomatosis peritonei 54 years after her initial surgery for an ovarian dermoid tumor. A separate teratomatous implant containing focally invasive adenocarcinoma was also present. CONCLUSION: This is by far the longest interval between initial diagnosis and identification of glial implants reported. Additionally, the presence of a separate malignant teratomatous implant suggests that teratomatous implants may retain malignant potential, in contrast to implants composed of purely benign glial tissue. PMID- 14751185 TI - Growing teratoma syndrome vs chemotherapeutic retroconversion: case report and review of the literature. AB - BACKGROUND: Immature ovarian teratoma is the third most common germ cell tumor (GCT) following dysgerminoma and endodermal sinus tumor. The treatment of choice during childbearing age for immature teratoma composes of unilateral oophorectomy and in case of metastatic disease postoperative chemotherapy (BEP). Finding a solid mass in the peritoneal or chest cavity during routine follow up raises the suspicion of distance recurrence. DiSaia was the first to describe the appearance of benign distant metastasis during routine follow up. He termed this phenomenon "chemotherapeutic retroconversion". Latter, Logothetis described what seems to be a similar phenomenon in testicular non-seminomatous germ cell tumor (NSGCT) that he called the "growing teratoma syndrome". CASE: We present a case of a 12-year old girl treated for growing teratoma syndrome after primary ovarian GCT. CONCLUSION: Review of the literature shows that this syndrome and the "chemotherapeutic retroconversion" are probably the same phenomenon. PMID- 14751186 TI - Low-grade myofibroblastic sarcoma of the vulva. AB - BACKGROUND: Vulvar sarcomas are rare tumors. A distinctive low-grade myofibroblastic sarcoma is described. CASE: A 46-year-old female presented with a progressively enlarging vulvar mass. Pathological evaluation revealed a low-grade myofibroblastic sarcoma. The immunophenotype is outlined and ultrastructural features are highlighted. Tumor cells were positive for p53 protein, smooth muscle actin, steroid receptors, and showed myofibroblastic differentiation on electron microscopy. The patient has been followed for >14 months without evidence of recurrence. CONCLUSION: The tumor was positive for p53, mitotically active, but was categorized as a low-grade malignancy. Immunohistochemical and ultrastructural criteria were utilized to distinguish this tumor from other neoplasms. PMID- 14751187 TI - Matched unrelated donor bone marrow transplantation for the treatment of platinum refractory ovarian carcinoma: a case report. AB - BACKGROUND: We are reporting on a case of platinum-refractory low-grade ovarian cancer responding to treatment with an allogeneic bone marrow transplantation from an unrelated donor. CASE: The 37-year-old patient received a preparative regimen consisting of fludarabine 25 mg/m(2) on days -6, -5, -4, -3 and -2, melphalan 70 mg/m(2) on days -3 and -2, and thymoglobulin 2 mg/kg on days -3, -2 and -1. An unrelated HLA-compatible bone marrow was infused on day 0. Graft versus-host disease prophylaxis consisted of tacrolimus and methotrexate. The patient demonstrated complete donor chimerism. Serial CT scans of the abdomen and pelvis over the following 15 months showed a slow regression of the malignant lesion. CONCLUSION: Engraftment of an unrelated donor marrow was achieved in a patient with ovarian cancer and induced a tumor response. This suggests the presence of a graft-versus-tumor effect in ovarian cancer. Further study is underway. PMID- 14751188 TI - Oral altretamine used as salvage therapy in recurrent ovarian cancer. AB - BACKGROUND: Altretamine has reported efficacy in the treatment of recurrent ovarian cancer following platinum-based therapy. This report presents the cases of two long-term survivors with recurrent ovarian cancer given oral altretamine. CASES: Two patients diagnosed with stage IIIC ovarian cancer underwent optimal cytoreductive surgery. Both women were subsequently treated with platinum-based chemotherapy. One had persistent cancer documented 2 months post therapy, while the other was disease-free for 22 months before recurring. Both received altretamine in a salvage setting. Each of these women achieved a prolonged response to third-line altretamine therapy, and one of whom was disease-free for 4 years and the other remains disease-free over 7 years following initiation of salvage therapy. CONCLUSION: Outpatient-administered oral altretamine can provide a prolonged disease-free interval with minimal toxicity. PMID- 14751189 TI - Primary ovarian undifferentiated non-small cell carcinoma, neuroendocrine type. AB - BACKGROUND: Non-small cell neuroendocrine carcinoma (NSCNEC) of the ovary is a rare aggressive neoplasm, characteristically arising in association with a surface epithelial tumor. CASE: A 27-year-old woman with no significant past medical history presented with a 17-cm left ovarian tumor. She underwent an exploratory laparotomy with resection of the pelvic mass. A diagnosis of stage IA non-small cell neuroendocrine carcinoma was rendered. She was subsequently treated with Taxol and Carboplatinum. CONCLUSION: This is a report of a rare primary ovarian non-small cell neuroendocrine carcinoma. In contrast to prior reports, this neoplasm was not associated with a surface epithelial tumor. PMID- 14751190 TI - Stage II-B carcinoma of the cervix complicated by bilateral pelvic kidneys. AB - BACKGROUND: The simultaneous occurrence of advanced cervical cancer and a pelvic kidney is uncommonly reported. It is an interesting clinical problem because the pelvic kidney lies within the radiation field. CASE: A patient found to have bilateral pelvic kidneys and stage II-B cervical cancer underwent an anterior exenteration. This was complicated by urinary conduit leakage and eventual nephrectomy for hemorrhage. She has been followed for 14 months with normal renal function and no evidence of disease. CONCLUSION: A primary surgical approach to stage II-B cervical carcinoma was offered to this patient because of bilateral pelvic kidneys. Anastomotic breakdown with urinary leakage after an ileal conduit remains a significantly morbid problem. PMID- 14751191 TI - Verrucous carcinoma of the vulva in Turner syndrome. AB - BACKGROUND: Verrucous carcinoma of the vulva in a patient with Turner syndrome is extremely rare. CASE REPORT: A 7.5 x 4 cm exophytic tumor of the vulva with a palpable ipsilateral groin lymph node developed in a 41-year-old single nulligravid Japanese woman with short stature and a history of primary amenorrhea. Analysis of karyotype demonstrated mosaic Turner syndrome. She underwent total deep vulvectomy with en-block inguinal femoral lymphadenectomy. Histopathological finding of the extirpated specimen demonstrated verrucous carcinoma with clear margins and the absence of metastasis to the lymph nodes. There was no sign of HPV genome detected in the carcinoma tissue. CONCLUSION: This seems to be the first case of vulvar verrucous carcinoma in a patient with Turner syndrome. PMID- 14751192 TI - Survival after relapse in patients with endometrial cancer: results from a randomized trial. C.L. Creutzberg, W.L.J. van Putten, P.C. Koper, et al., Gynecologic Oncology 89 (2003) 201-209. PMID- 14751194 TI - Commenting on "HER2/neu overexpression: has the Achilles' heel of uterine serous papillary carcinoma been exposed? (88:263-5) by Santin AD". PMID- 14751199 TI - Our right to work, our demand to be heard: people with disabilities, the 2004 election, and beyond. PMID- 14751200 TI - Focal network involvement in generalized seizures: new insights from electroconvulsive therapy. AB - Generalized seizures are commonly thought to involve the entire brain homogeneously. However, recent evidence suggests that selective cortical subcortical networks may be crucial for the initiation, propagation, and behavioral manifestations of generalized seizures, while other brain regions are relatively spared. Here we review previous studies, and describe a new human model system for the investigation of generalized seizures: single-photon emission computed tomography, ictal-interictal difference imaging of generalized tonic-clonic seizures induced by electroconvulsive therapy (ECT). Bitemporal ECT activates focal bilateral frontotemporal and parietal association cortex, sparing other regions; bifrontal ECT activates mainly prefrontal cortex; while in right unilateral ECT the left frontotemporal region is relatively spared. Associated midline subcortical networks are also involved. Focal verbal memory deficits parallel the focal regions involved in these neuroimaging studies. Further studies of this kind may elucidate specific networks in generalized tonic-clonic seizures, providing targets for new therapeutic interventions in epilepsy. PMID- 14751201 TI - Paradoxical results in the cure of chronic illness: the "burden of normality" as exemplified following seizure surgery. AB - Little has been written about processes of recovery following life-changing medical interventions for chronic illness. This article reviews our research with chronic epilepsy patients undergoing neurosurgery for the relief of intractable partial seizures. This research has given rise to a new conceptualization of adjustment and outcome following effective treatment of chronic illness, representing the first, detailed characterization of this process from a psychological and psychosocial perspective. Crucial to outcome are patient and family expectations prior to treatment, and learning to discard roles associated with chronic illness after treatment. These and the posttreatment affective functioning of the patient temper the view of medical outcome, and can account for paradoxical clinical effects, such as worsening patient psychosocial functioning in the context of medical treatment success. Our results have clear implications for the clinical management of chronically ill patients and their families to optimize treatment outcome. PMID- 14751202 TI - Differences in memory performance and other clinical characteristics in patients with mesial temporal lobe epilepsy with and without hippocampal atrophy. AB - Mesial temporal lobe epilepsy (MTLE) is usually accompanied by memory deficits due to damage to the hippocampal system. In most studies, however, the influence of hippocampal atrophy (HA) is confounded with other variables, such as: type of initial precipitating injury and pathological substrate, effect of lesion (HA) lateralization, history of febrile seizures, status epilepticus, age of seizure onset, duration of epilepsy, seizure frequency, and antiepileptic drugs (AEDs). To investigate the relationship between memory deficits and these variables, we studied 20 patients with MTLE and signs of HA on MRI and 15 MTLE patients with normal high-resolution MRI. The findings indicated that (1) HA, earlier onset of seizures, longer duration of epilepsy, higher seizure frequency, and AEDs (polytherapy) are associated with memory deficits; and (2) there is a close relationship between deficits of verbal memory and left HA, but not between visual memory and right HA. PMID- 14751203 TI - Pharmacologic evidence for a parasympathetic role in seizure-induced neurocardiac regulatory abnormalities. AB - PURPOSE: We evaluated whether postictal cardiac arrhythmia can be prevented by pharmacologic blockage of peripheral muscarinic receptors in an experimental model of epilepsy in rats. METHODS: Rats were prepared for chronic electrocardiograph recording and pretreated with atropine methyl bromide (2 or 10mg/kg i.p.) or saline prior to exposure to maximal electroshock (MES). The resulting seizure severity and duration of cardiac arrhythmia were measured. RESULTS: Atropine methyl bromide did not significantly affect seizure severity in comparison to control animals but reduced the arrhythmia at a dose of 2mg/kg, and completely suppressed arrhythmia at 10mg/kg. CONCLUSIONS: Postictal arrhythmia following MES-induced seizures may be blocked by pretreatment with atropine methyl bromide, a peripherally acting parasympatholytic agent. Our findings support previous observations that suggest strong participation of the parasympathetic system in postictal arrhythmia. This may be important for clinical suppression of cardiac arrhythmia in persons with uncontrolled epilepsy, who are at risk for sudden unexpected death. PMID- 14751204 TI - Long-term tolerability of lamotrigine: data from a 6-year continuation study. AB - This open-label, 6-year continuation study of several short-term clinical trials was conducted to assess the long-term tolerability and efficacy of lamotrigine when used as adjunctive therapy or monotherapy for partial seizures in adult patients (> or =16 years) with epilepsy. Study visits occurred every 24 weeks throughout the treatment period. Of the 527 patients enrolled in the long-term continuation study, 508 were exposed to lamotrigine for at least 6 months (including their exposure in the primary clinical study), and 248 were exposed to lamotrigine for at least 5 years. Of the 527 patients, 75 received initial lamotrigine exposure during this study. Investigators judged that overall clinical status at the end of the study or at time of discontinuation (whichever occurred first) was improved moderately or markedly relative to prelamotrigine clinical status for 36% of patients. The most common treatment-emergent adverse events (regardless of suspected cause) were dizziness, diplopia, and headache. The only serious treatment-emergent adverse event occurring at a frequency exceeding 2% was accidental injury (2.7% of patients). Adverse events prompted 28 patients to discontinue from the study. The most common adverse events leading to discontinuation were dizziness (1.3%), headache (0.8%), rash (0.8%), and somnolence (0.6%). All adverse events resolved without sequelae. Lamotrigine administered as monotherapy or adjunctive therapy during a 6-year open-label continuation study was associated with a low incidence of adverse events in adult patients with epilepsy. PMID- 14751205 TI - Cognitive and psychological predictors of everyday memory in children with intractable epilepsy. AB - Children with epilepsy have known deficits on objective measures of learning and memory. Parents and children report that memory deficits have a negative impact on everyday functioning. In adults with epilepsy, self-report of memory is more strongly associated with depression than performance on memory tests. We investigated the cognitive and psychological predictors of everyday memory in 37 children with medically intractable epilepsy, using children's self-report and parent ratings of everyday memory performance and standard tests of attention, intelligence, visual and verbal memory, working memory, and mood/emotional state. Standard multiple regressions demonstrated that only a parent report measure of attention uniquely and significantly (P< or =0.05) predicted estimates of everyday memory performance, accounting for 33% of variance in children's own ratings and 27% of variance in parents' ratings. Findings suggest that everyday memory in children with intractable epilepsy differs from that of adults; attentional problems may underlie everyday memory problems in these children. PMID- 14751206 TI - Day/night patterns of focal seizures. AB - PURPOSE: In many patients with epilepsy seizures occur with a day/night pattern. Our aims were to compare day/night patterns in seizure frequency among patients with different epileptogenic regions. METHODS: We analyzed video-EEG recordings in 15 patients with temporal lobe (TLE) and 11 with extratemporal lobe epilepsy (XTLE). Each seizure was classified according to subject group (TLE vs XTLE), sleep/wake state, and time of day of seizure occurrence (grouped into 6 x 4-hour "bins"). RESULTS: Of 90 seizures, 41 occurred in TLE and 49 in XTLE patients. There were day/night patterns of seizure occurrence in each group, with differences in the patterns between groups. In TLE, 50% of seizures occurred between the hours of 15:00 and 19:00 (17% would be expected by chance in each 4 hour "bin": F=3.59, P<0.006). In XTLE, there was a peak between 19:00 and 23:00 (47%: F=4.72, P<0.0018). The effect of time on seizures was least pronounced in the XTLE patients who had more than one epileptogenic region. The proportion of seizures occurring from sleep was significantly less in TLE (19%) than in XTLE patients (41%) [P<0.04, Fisher's exact test]. CONCLUSIONS: There are clear day/night patterns of seizure occurrence in epilepsy, with differences in the patterns between TLE and XTLE. There is an additional interaction with sleep/wake state, with relatively few seizures occurring from sleep in TLE compared with XTLE. Thus, it appears that both sleep/wake state and day/night or circadian rhythms may affect seizure proclivity, with different effects depending on the location of the epileptogenic region. PMID- 14751207 TI - Seizure disorders, depression, and health-related quality of life. AB - Disease and mood states are important determinants of quality of life (QOL). Low QOL, due to mood states, can be expected especially in psychiatric disorders such as depression. However, patients with seizure disorders may be even more affected because of the combined burden of physical episodes, psychiatric comorbidities, and psychosocial factors (e.g., stigma). In this study, we compare the quality of life in seizure disorders and clinical depression. Based on our earlier findings, we hypothesize that epilepsy patients fare better than patients with psychogenic, nonepileptic seizures (PNES), and we speculate that QOL in PNES is also lower relative to clinical depression. We estimate the relationships between type of seizures (epilepsy vs PNES), depression, and QOL (SF-36) using multiple regression, and we compare the SF-36 scores of patients with epilepsy and PNES (n=194) with the normative data for clinical depression using one-sample t tests. Our findings indicate that depression contributes to the poor QOL in both epilepsy and PNES, but the patients with PNES, even those without depression, have worse QOL compared with both the epilepsy patients and the depression norms. We conclude that evaluating and treating mood states is as important as treating PNES itself when caring for patients with PNES, and it might be the first step toward improving their QOL. PMID- 14751208 TI - Exploring the evaluation of antiepileptic drug change in people with intellectual disabilities and high-frequency epileptic seizures: seizure control and sustained responsiveness to the environment. AB - PURPOSE: Optimum antiepilepsy medication should be successful in reducing seizures with minimal adverse effects on the patient's ability to concentrate or general level of awareness. The purpose was to investigate the potential of a method of measuring responsiveness to environmental events as a means of reflecting awareness levels among people with intellectual disabilities undergoing review of medication for high-frequency epileptic seizures. METHODS: Observations of 22 participants referred to a specialist clinic were conducted three times a month over a 5-month period following the initial baseline measures and clinical intervention. Behavioral responsiveness was measured by calculating the likelihood of appropriate activity occurring given the occurrence of staff interaction. This likelihood was represented by the statistic Yule's Q. Seizure frequency was also evaluated. RESULTS: Participant responsiveness after drug review was similar to baseline indicating an absence of long-term adverse effects. Participants experienced a significant decrease in seizure frequency. CONCLUSION: It was concluded that drug review led to seizure reduction while behavioral measurement confirmed no loss of responsiveness. PMID- 14751209 TI - Pharmacological outcomes in people with mental retardation and epilepsy. AB - The aim of this study was to examine prospective outcomes in mentally retarded people with epilepsy seen for the first time during 1998 until the end of 2001. Two hundred and fourteen patients (120 men, 94 women, aged 11-70 years [median 34 years]) were referred over this 4-year period. Median duration of follow-up was 18 months (range, 13-36 months). Seventeen (8%) had nonepileptic attacks only, 10 of whom were being treated with antiepileptic drugs (AEDs). The remaining 197 (92%) patients had epilepsy, the majority (n=151, 77%) presenting with focal seizures. A total of 22 patients were started on AED treatment, with seizure freedom for at least a year being achieved in 10 (45%). AED manipulation was undertaken in a further 136 patients, resulting in 59 (43%) becoming seizure free. No relationship was found between extent of mental retardation and seizure control. There was no deterioration in mean caregiver scores rating sleep, appetite, alertness, and behavior. These findings suggest that AED therapy can result in seizure freedom in more than 40% of mentally retarded people with epilepsy without producing unacceptable toxicity. PMID- 14751210 TI - Effects of epilepsy surgery on quality of life: a controlled study in a Middle Eastern population. AB - The purpose of this study was to investigate which areas of quality of life (QOL) change after epilepsy surgery and generate QOL data specific to the Lebanese population. The QOL of 20 consecutive patients 1 year after surgery was compared (using the ESI-55 scale) with that of 20 matched patients who underwent the same presurgical evaluation, but no surgery. Overall QOL improved in both groups, but was greater in the surgery group (85% seizure free as compared to 0% in nonsurgery group). Significant differences were noted within the "well-being" domain including health perception (5/9 items), energy-fatigue (2/4), and emotional well-being (2/5). Differences were less common in the "functioning" domain including physical (1/10 items), social (1/2), and cognitive (0/5) functioning scales, and in the role limitation domain (1/17 items). Our patients experienced, 1 year after epilepsy surgery, improvements similar to those reported in Western populations after a similar period: they had marked improvements in overall QOL, health perception, well-being, and cognitive functioning areas. They had less remarkable improvements in social functioning and role limitation areas. PMID- 14751211 TI - Epilepsy surgery: patient-perceived long-term costs and benefits. AB - PURPOSE: The goal of this study was to assess the patient-perceived costs and benefits associated with the longer-term outcomes of epilepsy surgery in patients who underwent anterior temporal lobectomy or selective amygdalohippocampectomy. METHODS: Surgery patients who were assessed in 1997 were reassessed in 2003. Demographic, clinical, and psychosocial details were collected using a validated self-completion questionnaire. Data were collected from 67 patients who had undergone surgery. RESULTS: Forty-five percent were seizure-free. There were significant differences (P<0.001) between the seizure-free (SF) and continuous seizure (CS) groups with respect to anxiety, depression, impact of epilepsy, self esteem, mastery, stigma, affect balance, self-reported health, and quality of life. More SF patients were also employed and driving (P<0.001). Despite these differences there were no differences for regret over surgery but there were differences for satisfaction and success ratings. CONCLUSIONS: Patients who were not SF, in the longer term, had little regret undergoing surgery but were less likely to be satisfied and had a poorer psychosocial profile. PMID- 14751212 TI - Psychogenic nonepileptic seizures are associated with an increased risk of obesity. AB - Psychogenic nonepileptic seizures (NES) are somatic manifestations of psychological distress. There is some evidence that weight problems are more common in patients with psychiatric illness. We have observed that patients admitted for video-EEG monitoring who we diagnosed with NES commonly have a larger body habitus than patients with epilepsy. The goal of this study was to test our hypothesis that there was a significant difference in body mass index (BMI) in patients with nonepileptic seizures compared with their epileptic counterparts. We compared the BMIs of 46 NES patients and 46 age- and gender matched epileptic controls and found that the NES patients had significantly higher BMIs (30.5 vs 26.1, P=0.006) than controls. This remained true after controlling for weight-gain properties of antiepileptic drugs. These results are compared with the prevalence of overweight and obesity in the general population. Possible explanations of the findings and limitations of the study are discussed. PMID- 14751213 TI - Effect of interstitial stereotactic radiosurgery on behavior and subjective handicap of epilepsy in patients with gelastic epilepsy. AB - Patients with symptomatic epilepsy due to hypothalamic hamartomas often are compromised not only by pharmacoresistant epileptic seizures but also by behavioral disturbances and cognitive dysfunction. We report the effect of successful treatment with stereotactic interstitial radiosurgery by intrahypothalamic implantation of 125I seeds on behavior and subjective handicap. In all patients rendered seizure-free or suffering only from auras, improvement of behavior was reported by parents and colleagues or schoolteachers. Parents' ratings according to the Child Behavior Checklist showed improvements with respect to social problems and attention. Self-ratings of quality of life by adult patients showed improvements in activities, working situation, and self perception. These improvements were not observed in patients in whom clinically manifest seizures and interictal EEG discharges persisted after radiosurgery. PMID- 14751214 TI - Concerns regarding lamotrigine and breast-feeding. AB - PURPOSE: Many women with epilepsy who are planning a pregnancy are treated with lamotrigine (LTG), resulting in greater fetal exposure to the drug. Current care guidelines suggest that mothers with epilepsy breast-feed their children. These recommendations are made without regard to how nursing newborns metabolize medication. Lamotrigine is extensively metabolized by glucuronidation, which is immature in neonates and may lead to accumulation of medication. This article reports LTG levels in full-term nursing newborns born to mothers with epilepsy on lamotrigine monotherapy. METHODS: Serum LTG levels were obtained in nursing mothers and their neonates on Day 10 of life. Maternal LTG clearance during pregnancy and postpartum was determined and correlated with levels. RESULTS: Four mothers with partial epilepsy on LTG monotherapy were evaluated. Serum LTG levels in nursing newborns ranged from <1.0 to 2.0 microg/mL on Day 10 of life. Three babies had LTG levels >1.0 microg/mL. After excluding one child with an undetectable level, the LTG levels in newborns were on average 30% (range 20-43%) of the maternal drug level. No decline was noted in two children with repeat levels at 2 months. CONCLUSION: Serum concentrations of LTG in breast-fed children were higher than expected, in some cases reaching "therapeutic" ranges. These high levels may be explained by poor neonatal drug elimination due to inefficient glucuronidation. Our observation that not all newborns had a high LTG level suggests considerable genetic variability in metabolism. Our limited data suggest monitoring blood levels in nursing children and the need for individual counseling for women with epilepsy regarding breast-feeding. PMID- 14751215 TI - Face and facial expression memory in temporal lobe epilepsy patients: preliminary results. AB - Right temporal lobe structures are involved in face and facial expression processing and in mnestic functions. Face and facial expression memory was investigated in 15 patients with left (LTLE) and 18 patients with right (RTLE) temporal lobe epilepsy as well as 13 healthy controls. Pairs of pictures combining four faces and four emotions had to be matched according to face identity or facial expression. In the memory tasks, the two pictures of a pair were divided by a memory interval of 2000 milliseconds, whereas in the perception tasks (control condition) both pictures were presented simultaneously. RTLE patients had significantly lower scores than healthy controls in face memory. LTLE patients had significantly lower scores than healthy controls in face and facial expression memory. The data confirm impaired face memory in RTLE patients and show that LTLE patients display deficits in face as well as in facial expression memory. Results are discussed according to functional reorganization, memory strategies, perception performance, naming problems, and group characteristics. PMID- 14751216 TI - Obsessive-compulsive disorder after epilepsy surgery. AB - It is a well-known fact that after epilepsy surgery (ES) preexisting psychopathology may deteriorate or de novo psychopathological syndromes, mainly of a depressive and psychotic nature, may appear. Previously, recovery of obsessive-compulsive disorder (OCD) after ES has been reported in patients who had comorbid OCD preoperatively; however, there have been no reports on the appearance of de novo OCD interfering with daily living activities post-ES. This is the first report of OCD after ES in patients with mesial temporal lobe epilepsy (MTLE). Five patients with MTLE were identified with obsessive personality traits before surgery. Within the first 2 months after ES, two of these MTLE patients fulfilled OCD diagnostic criteria. These OCD patients were not any different from the other three patients with respect to age, age of onset of epilepsy, seizure types, and seizure frequency. All patients stopped having seizures postoperatively, but the OCD patients had worse quality of life postoperatively than preoperatively. Our findings show that those patients with obsessive traits preoperatively should be carefully monitored after ES. PMID- 14751217 TI - The neuromatrix and the epileptic brain: behavioral and learning preservation in limbic epileptic rats treated with ketamine but not acepromazine. AB - Conceiving the organization of the brain as a "neuromatrix" could provide significant insights into how different injuries to the nervous system could result in very distinct changes in behavior. The use of different pharmacological treatments to combat the deleterious consequences of such injuries is common practice. However, such treatments may have the capacity to alter the configurations of various neuronal circuits that contribute to the "neuromatrix" by selectively preventing damage to some pathways while facilitating the spread of destruction along others. The choice of pharmacological treatment may have profound consequences on the recovery of normal functioning following injury. We examined the behavior of rats treated with one of two potentially neuroprotective agents, the N-methyl-D-aspartate antagonist ketamine and the atypical neuroleptic acepromazine, on seizures induced by lithium-pilocarpine. Rats treated with ketamine following seizure onset were virtually indistinguishable from nonepileptic controls on a variety of behavioral tasks that included tests on learning, memory, and anxiety. In contrast, acepromazine-treated rats showed marked deficits on all learning and behavioral measures tested. These results suggest that administration of ketamine relatively soon after the emergence of epilepsy can prevent many of the cognitive deficits that are commonly found in rats subjected to lithium-pilocarpine-induced seizures. Further clinical testing investigating ketamine as a potential adjunct treatment for epilepsy may be well warranted. PMID- 14751219 TI - Excessive masturbation after epilepsy surgery. AB - Sexual behavior changes as well as depression, anxiety, and organic mood/personality disorders have been reported in temporal lobe epilepsy (TLE) patients before and after epilepsy surgery. The authors describe a 14-year-old girl with symptoms of excessive masturbation in inappropriate places, social withdrawal, irritability, aggressive behavior, and crying spells after selective amygdalohippocampectomy for medically intractable TLE with hippocampal sclerosis. Since the family members felt extremely embarrassed, they were upset and angry with the patient which, in turn, increased her depressive symptoms. Both her excessive masturbation behavior and depressive symptoms remitted within 2 months of psychoeducative intervention and treatment with citalopram 20mg/day. Excessive masturbation is proposed to be related to the psychosocial changes due to seizure free status after surgery as well as other possible mechanisms such as Kluver Bucy syndrome features and neurophysiologic changes associated with the cessation of epileptic discharges. This case demonstrates that psychiatric problems and sexual changes encountered after epilepsy surgery are possibly multifactorial and in adolescence hypersexuality may be manifested as excessive masturbation behavior. PMID- 14751218 TI - Vagus nerve stimulation for pharmacoresistant epilepsy: clinical symptoms with end of service. AB - PURPOSE: Limited capability exists to predict when vagus nerve stimulation (VNS) battery deterioration becomes significant. Initial models last 2-5 years. We evaluated the first 18 patients with pharmacoresistent epilepsy after reimplantation to examine the clinical course observed during VNS end of service (EOS). METHODS: Of 72 patients with VNS, 18 patients had generator replacement. EOS was estimated based on duration of use and stimulus parameters in accordance with manufacturer guidelines. Eight males and ten females had pharmacoresistent epilepsy for a mean of 17.9 years. Thirteen with localization-related epilepsy (LRE) and 5 nonverbal patients with symptomatic generalized epilepsy (SGE) failed a mean of 11.1 antiepileptic drugs (AEDs) over 21.5 years. Seven had intracranial evaluations and five failed epilepsy surgery. Reimplantation was performed after a mean of 34.4 months. Symptoms at end of service (EOS) were addressed by postoperative survey submitted at initial reprogramming within 2 weeks of reimplantation. Stimulus parameters were compared before and after surgery. RESULTS: Nine of thirteen (69.2%) verbal patients and 11 of 18 (61.1%) total patients had signs or symptoms prior to replacement, suggesting clinical EOS, and 4 of 18 (22.2%) failed interrogation denoting battery failure without symptoms; however, this did not reach significance (chi2=0.359,p=0.54). Increased seizures were the most frequent sign in 8 of 18 (44.4%), with intensification in 7 of 18 (38.9%). Irregular stimulation was detected in 5 of 18 (27.7%), with less intense stimulation in 4 of 18 (22.2%). Painful stimulation and behavioral worsening each occurred in 2 of 18 (11.1%). A subjective improvement in function after reimplantation was noted in 12 of 13 (92.3%) verbal patients, with greater intensity and consistency. Maximally tolerated reimplant current averaged -0.56 mA less. All but one (94.4%) felt surgery should be performed before clinical EOS occurred. CONCLUSIONS: We conclude that clinical signs and symptoms may arise during VNS EOS and following replacement. Seizure increase or a change in seizure pattern was most frequently observed. The tolerated reimplant current was less than the preoperative output current in most cases. Battery replacement before EOS appears desirable from a patient perspective. PMID- 14751220 TI - Successful resonance Raman study of cresolase activity of mushroom tyrosinase. AB - Mono-oxygenase (cresolase) activity of mushroom tyrosinase (MT) in the presence of 4-[(4-hydroxyphenyl)azo]-benzenesulfonamide (HPABS) was successfully studied by resonance Raman (rR) spectroscopy. HPABS is a synthetic competitive inhibitor (K(i)=7.17 x 10(-6)M) for the cresolase activity with a large extinction coefficient at 365 nm. Upon reacting with MT, HPABS produced an enzyme-inhibitor (EI) complex with sufficiently long life span. Analyzing the ensuing spectrum indicates that the azo tautomer of HPABS binds to the enzyme and retains its geometrical isomeric form in the EI complex. The observed changes in the rR spectrum of HPABS after binding to MT support the idea that an electrophilic attack on the inhibitor has happened. Similar experiments were designed for studying the oxidase activity of MT. However, the enzymatic reaction, even in the presence of 4-[(2,4-dinitrophenyl)azo]-1,2-benzenediols was still fast enough to tan the reaction solution quickly and render its rR spectrum impregnable background. PMID- 14751221 TI - Antibody detection of SARS-CoV spike and nucleocapsid protein. AB - Early detection and identification of SARS-CoV-infected patients and actions to prevent transmission are absolutely critical to prevent another SARS outbreak. Antibodies that specifically recognize the SARS-CoV spike and nucleocapsid proteins may provide a rapid screening method to allow accurate identification and isolation of patients with the virus early in their infection. For this reason, we raised peptide-induced polyclonal antibodies against SARS-CoV spike protein and polyclonal antibodies against SARS-CoV nucleocapsid protein using 6x His nucleocapsid recombinant protein. Western blot analysis and immunofluorescent staining showed that these antibodies specifically recognized SARS-CoV. PMID- 14751222 TI - Indirect oxidation of 6-tetrahydrobiopterin by tyrosinase. AB - 6-Tetrahydrobiopterin is known to bind to an allosteric site of tyrosinase to directly inhibit the enzyme. However, simultaneous measurements of ultraviolet visible absorption spectra and oxygen consumption led us to conclude that the inhibition was due to oxidation of 6-tetrahydrobiopterin by dopaquinone. Immediately after addition of 6-tetrahydrobiopterin, tyrosinase stopped producing dopachrome from either tyrosine or dopa. Duration of inhibition was proportional to the concentration of added 6-tetrahydrobiopterin and the enzyme activity was fully restored after the inhibition. Surprisingly, there was a rapid consumption of oxygen during the inhibition period. In addition, absorption spectra indicated that the only reaction that occurred during the inhibition was oxidation of 6 tetrahydrobiopterin to 7,8-dihydrobiopterin. In the absence of tyrosine or dopa, tyrosinase did not oxidize 6-tetrahydrobiopterin, suggesting that a reaction intermediate between dopa and dopachrome was a target for the inhibition. We propose a new mechanism in which dopa is oxidized to dopaquinone and the latter, instead of producing dopachrome, is reduced back to dopa by 6 tetrahydrobiopterin. PMID- 14751223 TI - Characterization of a cytotoxic pilin subunit of Xenorhabdus nematophila. AB - Xenorhabdus nematophila is an insect pathogenic bacterium, known to produce protein toxins that kill the larval host. We have described a cytotoxic pilin subunit of X. nematophila, which is expressed on the cell surface and also secreted in the extracellular medium associated with outer membrane vesicles. A 17kDa pilin subunit was isolated and purified from X. nematophila cell surface. The protein showed cytotoxicity to larval hemocytes of Helicoverpa armigera in an in vitro assay, causing agglutination of the cells, and releasing cytoplasmic enzyme lactate dehydrogenase in the medium. The pilin protein was able to bind to the surface of larval hemocytes. The binding and cytotoxicity of the purified 17kDa protein to hemocytes was inhibited by antiserum raised against the pilin protein. The study demonstrates for the first time a cytotoxic structural subunit of pilin from an entomopathogenic bacterium X. nematophila that is excreted in the extracellular medium with outer membrane vesicles. PMID- 14751224 TI - Rapid method for mean telomere length measurement directly from cell lysates. AB - Telomere length is involved in cell survival, tumorigenesis, and early aging. We present here an innovative method to determine the mean telomere length without any DNA purification. Our strategy is to measure both the DNA concentration and the number of telomeric units (TTAGGG) directly from cell lysate produced by the combined action of NaOH (pH>13) and sonication directly on cell pellet. Telomere units are quantified using an enzyme hybridization assay on 96-well microtiter plates grafted with a captor sequence. A biotin-coupled-tracer oligonucleotide hybridizes with telomere fragments and the enzymatic reaction is performed with a streptavidin-acetylcholinesterase conjugate, using the colorimetric method of Ellman. OD measure is directly proportional to the number of telomere units in cell lysate. This scalable technique allows the determination of mean telomere length simultaneously in many samples. This assay will be highly efficient to screen new drugs involved in chemotherapy targeting telomerase or directly telomeres. PMID- 14751225 TI - Anaerobic transformation of 2,4,6-TNT by bovine ruminal microbes. AB - Degradation of TNT by bovine rumen fluid, a novel source of anaerobic microbes, was investigated. Whole rumen fluid contents were spiked with TNT and incubated for a 24h time period. Supernatant samples taken at 0, 1, 2, 4, and 24h were analyzed by reverse-phase HPLC with diode array detection. Within 1h, TNT was not detectable and reduction products of TNT including 2-hydroxyl-amino-4,6 dinitrotoluene, 4-hydroxylamino-2,6-dinitrotoluene, and 4-amino-2,6 dinitrotoluene were present with smaller amounts of diamino-nitrotoluenes. Within 2h, only the diamino and dihydroxyamino-nitrotoluene products remained. After 4h, 2,4-diamino-6-nitrotoluene and 2,4-dihydroxyamino-6-nitrotoluene were the only known molecular species left. At 24h known UV absorbing metabolites were no longer detected, suggesting further transformation such as complete reduction to triaminotoluene or destruction of the aromatic ring of TNT may have occurred. TNT was not transformed at 24h in autoclaved and buffered controls. This study presents the first direct evidence of biodegradation of TNT by ruminal microbes. PMID- 14751226 TI - Dimerization modulates the activity of the orphan nuclear receptor ERRgamma. AB - Estrogen-related receptor gamma (ERRgamma) is an orphan nuclear receptor lacking identified natural ligands. However, 4-hydroxytamoxifen and diethylstilbestrol were recently shown to bind to and inhibit ERRgamma activity. ERR activates transcription constitutively as a monomer. We show here that ERRgamma forms also dimers via its ligand-binding domain. Homodimerization enhances the transcriptional activity. In contrast, heterodimerization with the related receptor ERRalpha inhibits the activities of both ERRgamma and ERRalpha. The inverse ERRgamma agonist 4OHT further inhibits the activity of the ERRgamma ERRalpha heterodimer, indicating that 4OHT may modulate ERRalpha signaling via ERRgamma. Receptor dimerization thus modulates the transcriptional activities of ERRs. PMID- 14751227 TI - Deletion of the cytoplasmic domain increases basal shedding of angiotensin converting enzyme. AB - Ectodomain shedding generates soluble isoforms of cell-surface proteins, including angiotensin-converting enzyme (ACE). Increasing evidence suggests that the juxtamembrane stalk of ACE, where proteolytic cleavage-release occurs, is not the major site of sheddase recognition. The role of the cytoplasmic domain has not been completely defined. We deleted the cytoplasmic domain of human testis ACE and found that this truncation mutant (ACE-DeltaCYT) was shed constitutively from the surface of transfected CHO-K1 cells. Phorbol ester treatment produced only a slight increase in shedding of ACE-DeltaCYT, unlike the marked stimulation seen with wild-type ACE. However, for both wild-type ACE and ACE-DeltaCYT, shedding was inhibited by the peptide hydroxamate TAPI and the major cleavage site was identical, indicating the involvement of similar or identical sheddases. Cytochalasin D markedly increased the basal shedding of wild-type ACE but had little effect on the shedding of ACE-DeltaCYT. These data suggest that the cytoplasmic domain of ACE interacts with the actin cytoskeleton and that this interaction is a negative regulator of ectodomain shedding. PMID- 14751228 TI - Systematic screening of potential beta-cell imaging agents. AB - The beta-cell loss seen in diabetes mellitus could be monitored clinically by positron emission tomography (PET) if imaging agents were sufficiently specific for beta-cells to overcome the high ratio of non-beta-cell to beta-cell tissue in pancreas. In this report, we present a screening assay for identifying beta-cell specific compounds that is based on the relative accumulation and retention by islet, INS-1, and exocrine (PANC-1) cells of candidate molecules. Molecules thought to have a high affinity for beta-cells were tested and included glibenclamide, tolbutamide, serotonin, L-DOPA, dopamine, nicotinamide, fluorodeoxyglucose, and fluorodithizone. Glibenclamide and fluorodithizone were the most specific, but the specificity ratios fell well below those needed to attain robust signal to background ratio as a PET imaging agent for quantifying beta-cell mass. In vivo tests of the biodistribution of glibenclamide and fluorodithizone in rats indicated that the compounds were not specifically associated with pancreas, bearing out the predictions of the in vitro screen. PMID- 14751229 TI - A novel inhibitor protein of N-myristoyltransferase from Escherichia coli. AB - Myristoyl-CoA:protein N-myristoyltransferase (NMT) catalyzes the covalent attachment of myristate to the N-terminal of the glycine residue of various eukaryotic and viral proteins of diverse functions. Earlier, we have demonstrated that NMT activity is elevated in colon and gall bladder cancer. Attenuation of NMT activity may prove a novel therapeutic protocol for cancer. We report here a novel inhibitor protein of NMT being expressed in Escherichia coli cells containing the human NMT gene on increasing the incubation period from 5 to 24h. The inhibitor protein was purified by SP-Sepharose column chromatography, heat treatment, ammonium sulfate precipitation, and Superose 12 HR/30 FPLC column chromatography. The inhibitor protein had an apparent molecular mass of 10kDa by gel filtration. It inhibited human NMT in a concentration-dependent manner with 50% inhibition at 640+/-4.68nM. The inhibitor protein showed no direct interaction with myristoyl-CoA and demonstrated no demyristoylase or protease activity. Therefore, we conclude that the inhibitor protein acts directly on NMT. PMID- 14751230 TI - A mushroom (Ganoderma capense) lectin with spectacular thermostability, potent mitogenic activity on splenocytes, and antiproliferative activity toward tumor cells. AB - An 18-kDa lectin, with an N-terminal sequence displaying slight similarity to some lectins and fungal immunomodulatory proteins, was isolated from the mushroom Ganoderma capense (Lloyd) Teng. It exhibited more potent mitogenic activity than that of concanavalin A toward mouse splenocytes, and antiproliferative activity toward leukemia (L1210 and M1) cells and hepatoma (HepG2) cells. The isolation procedure entailed ion exchange chromatography on Q-Sepharose, fast protein liquid chromatography (FPLC)-ion exchange chromatography on Mono S, and FPLC-gel filtration on Superdex 75. D(+)-galactose and D(+)-galactosamine specifically inhibited the hemagglutinating activity of the lectin. The hemagglutinating activity of the lectin was not affected over the temperature range 0-100 degrees C and after exposure to 100 degrees C for 60min. The activity was stable in the pH range of 4-11, and after incubation with solutions of various chlorides (from 3.125 to 50mM) including NaCl, KCl, CaCl(2), MgCl(2), ZnCl(2), MnCl(2), and AlCl(3). However, it was potentiated by 12.5-50mM FeCl(3). The lectin was devoid of HIV-1 reverse transcriptase inhibitory and antifungal activities. PMID- 14751231 TI - Quantitative assessment of FGF regulation by cell surface heparan sulfates. AB - Heparin/heparan sulfate-like glycosaminoglycans (HSGAGs) modulate the activity of the fibroblast growth factor (FGF) family of proteins. Through interactions with both FGFs and FGF receptors (FGFRs), HSGAGs mediate FGF-FGFR binding and oligomerization leading to FGFR phosphorylation and initiation of intracellular signaling cascades. We describe a methodology to examine the impact of heparan sulfate fine structure and source on FGF-mediated signaling. Mitogenic assays using BaF3 cells transfected with specific FGFR isoforms allow for the quantification of FGF1 and FGF2 induced responses independent of conflicting influences. As such, this system enables a systematic investigation into the role of cell surface HSGAGs on FGF signaling. We demonstrate this approach using cell surface-derived HSGAGs and find that distinct HSGAGs elicit differential FGF response patterns through FGFR1c and FGFR3c. We conclude that this assay system can be used to probe the ability of distinct HSGAG species to regulate the activity of specific FGF-FGFR pairs. PMID- 14751232 TI - Functional crosstalk between Wnt signaling and Cdx-related transcriptional activation in the regulation of the claudin-2 promoter activity. AB - Assembly of tight junctions at the most apical part of the lateral cell membrane is a key event in the differentiation of polarized epithelial cells. Claudin-2, a transmembrane protein involved in tight junction strand formation, has turned out to play a crucial role for the paracellular barrier function by opening pores for small cations. Physiological and pathological variations of epithelial barrier function are accompanied by differential expression of tight junction proteins. Therefore, we characterized molecular mechanisms regulating claudin-2 gene expression. Genomic DNA containing the transcription start point of human claudin 2 was isolated and functionally characterized by reporter gene assays. Activity of the claudin-2 promoter was elevated in mouse mammary epithelial C57 cells expressing Wnt-1. LEF-1, a nuclear effector of the Wnt signaling pathway which is involved in the regulation of cell differentiation and polarization, was found to bind directly to the claudin-2 promoter as revealed by electrophoretic mobility shift assays. Expression of LEF-1 and beta-catenin both enhanced claudin-2 promoter activity. This increase was reduced after mutation of LEF-1 binding sites within the claudin-2 promoter. Furthermore, claudin-2 promoter activity was found to be enhanced by the TCF-4/beta-catenin transcription complex. Therefore, we conclude that gene expression mediated by the promoter of the human tight junction protein claudin-2 is regulated by factors involved in Wnt signaling. Moreover, a functional crosstalk between Wnt signaling and transcriptional activation related to caudal-related homeobox (Cdx) proteins could be demonstrated in the regulation of claudin-2 promoter-mediated gene expression. PMID- 14751233 TI - ADAMs, a disintegrin and metalloproteinases, mediate shedding of oxytocinase. AB - Placental leucine aminopeptidase (P-LAP), a type-II transmembrane protease responsible for oxytocin degradation during pregnancy, is converted to a soluble form through proteolytic cleavage. The goal of this study was to determine the nature of the P-LAP secretase activity. The hydroxamic acid-based metalloprotease inhibitors GM6001 and ONO-4817 as well as the TNF-alpha protease inhibitor-2 (TAPI-2) reduced P-LAP release, while tissue inhibitors of metalloproteinase (TIMP)-1 and TIMP-2, which are matrix metalloproteinase inhibitors, had no effect on P-LAP release in Chinese hamster ovary (CHO) cells stably overexpressing P LAP, thus indicating possible involvement of ADAM (a disintegrin and metalloproteinase) members in P-LAP shedding. Furthermore, overexpression of ADAM9 and ADAM12 increased P-LAP release in P-LAP-CHO transfectants. Immunohistochemical analysis in human placenta demonstrated strong expression of ADAM12 in syncytiotrophoblasts, while little expression of ADAM9 was detected throughout the placenta. Our results suggest ADAM members, at least including ADAM12, are involved in P-LAP shedding in human placenta. PMID- 14751234 TI - Direct measurement of Ca2+ concentration in the SR of living cardiac myocytes. AB - Although abnormal sarcoplasmic reticulum (SR) Ca(2+) handling may cause heart failure, there has been no method to directly measure Ca(2+) concentration in SR ([Ca(2+)](SR)) of living cardiomyocytes. We have measured [Ca(2+)](SR) by expressing novel fluorescent Ca(2+) indicators yellow cameleon (YC) 2.1, YC3er, and YC4er in cultured neonatal rat cardiomyocytes. The distribution of YC2.1 was uniform in the cytoplasm, while that of YC3er/YC4er, containing the signal sequence which recruits them to SR, showed reticular pattern and was co-localized with SERCA2a. The treatment with caffeine reversibly decreased the emission ratio (R) in YC3er/YC4er-expressing myocytes, and the treatment with ryanodine and thapsigargin decreased R irreversibly. During the contraction-relaxation cycle, R was changed periodically in the YC2.1- and YC3er-expressing myocytes, but its direction of the change was opposite. These results suggest that YC3er/YC4er were specifically localized and functioned in SR as a [Ca(2+)](SR) indicator. This technique would be useful to understand the function of SR in failing myocardium. PMID- 14751235 TI - Regulation of osteoclastogenesis by three human RANKL isoforms expressed in NIH3T3 cells. AB - Receptor activator of nuclear factor-kappaB ligand (RANKL) induces osteoclastogenesis by binding with the receptor, receptor activator of nuclear factor-kappaB in the presence of macrophage colony-stimulating factor. Three human RANKL isoforms, hRANKL1, hRANKL2, and hRANKL3, were identified. hRANKL1 was identical to previously reported RANKL and possessed intracellular, transmembrane, and extracellular domains, hRANKL2 did not have the intracellular domain, and hRANKL3 did not have the intracellular and transmembrane domains. When bone marrow macrophages were cultured with NIH3T3 cells expressing hRANKL1, osteoclasts were formed, but when cultured with NIH3T3 cells expressing hRANKL2 or hRANKL3, no tartrate resistant acid phosphatase-positive cell was observed. In the coculture system, coexpression of hRANKL3 with hRANKL1 significantly inhibited the formation of osteoclasts by hRANKL1, but coexpression of hRANKL2 with hRANKL1 did not affect the osteoclastogenesis by hRANKL1 significantly. These results suggest that the activity of osteoclastogenesis by hRANKL1 is regulated by the attenuator, hRANKL3. PMID- 14751236 TI - Inactivating mutations block the tumor necrosis factor-alpha-converting enzyme in the early secretory pathway. AB - The ectodomain of different transmembrane molecules is released by a proteolytic event known as shedding. The metalloprotease disintegrin proTNF-alpha converting enzyme (TACE) is responsible for the shedding of various proteins, including protransforming growth factor-alpha (proTGF-alpha) and amyloid-beta precursor protein (APP). Inactive TACE accumulates in the early secretory pathway of cell mutants (M1 and M2) defective in proTGF-alpha and APP shedding. Although previous evidences indicated that the component mutated in M1 and M2 cells is different from TACE, recent results show the existence of two heterozygous point mutations in TACE from M2 cells. Here, we show that wild-type TACE stably transfected in M2 cells is processed, transported to the cell surface, and rescues the proTGF-alpha and APP shedding-defective phenotype. Furthermore, M1 cells also express mutant TACE and transfection with wild-type TACE restores the wild-type phenotype. Therefore, different inactivating mutations result in the accumulation of TACE in the early secretory pathway, emphasizing the importance of the initial steps in the biosynthesis of TACE. PMID- 14751237 TI - Human DDB2 splicing variants are dominant negative inhibitors of UV-damaged DNA repair. AB - Damaged DNA-binding protein (DDB) is a heterodimer (DDB1 and DDB2), which is implicated in the repair of UV-irradiated DNA damage. Here we have identified four DDB2 variants from HeLa cells (D1-D4) that are generated by alternative splicing. Analysis of tissue distribution by RT-PCR indicates that D1 is the most highly expressed in human brain and heart. A DNA repair assay revealed that both D1 and D2 are dominant negative inhibitors. Electrophoresis mobility shift assays indicated that D1 and D2 are not part of the damaged DNA-protein complex. Co immunoprecipitation studies show that DDB2-WT interacts with D1 and itself. Nuclear import of DDB1 was less induced by transfection with D1 than WT. Based on these results, D1 and D2 are dominant negative inhibitors of DNA repair, which is probably due to disruption of complex formation between DDB1 and DDB2-WT and of DDB1 nuclear import. PMID- 14751238 TI - Insulin-like growth factor binding protein-3 stimulates phosphatidylinositol 3 kinase in MCF-7 breast carcinoma cells. AB - Insulin-like growth factor binding protein-3 (IGFBP-3) is the most abundant IGFBP in serum and other biological fluids. Apart from its capacity for specific and high-affinity binding to IGFs, it also has so-called "IGF-independent" activities that modulate cell proliferation and survival/apoptosis. However, the molecular elements of the IGFBP-3 signalling pathway remain obscure. In this study, we investigated the possible implication of phosphatidylinositol 3-kinase (PI 3 kinase) activity in MCF-7 breast carcinoma cells. In cells incubated with IGFBP 3, both total and insulin receptor substrate-1 (IRS-1)-associated PI 3-kinase activities were rapidly stimulated, with maximal effects after 3 and 10min of incubation, respectively. IGFBP-3-induced PI 3-kinase activity was unaffected by the state of IRS-1 tyrosine phosphorylation. Since IGFBP-3 failed to stimulate PI 3-kinase activity in MDA-MB 231 breast carcinoma cells, its effects in MCF-7 cells could be considered as cell-type-specific. Pertussis toxin abolished IGFBP 3-stimulation of PI 3-kinase activity, suggesting that this IGFBP-3 signalling pathway depends upon a pertussis toxin-sensitive G protein. Our results provide further evidence that IGFBP-3 directly triggers a specific intracellular signal in MCF-7 cells. PMID- 14751240 TI - Effects of C-reactive protein on atherogenic mediators and adrenomedullin in human coronary artery endothelial and smooth muscle cells. AB - We examined the effects of recombinant human C-reactive protein (rhCRP) on atherosclerosis-related factors in cultured human coronary artery endothelial and smooth muscle cells (HCAECs and HCASMCs). After removing endotoxin from commercial rhCRP preparations using the appropriate column, the purified (P) rhCRP retained the ability to Ca(2+)-dependently bind to phosphorylcholine, but did not augment the secretion of interleukin-6 and MCP-1 from HCAECs, as non purified (NP)-rhCRP did. By contrast, P-rhCRP elicited 2- to 3-fold increases in the secretion of both hormones from HCASMCs, though the effect was smaller than that obtained with NP-rhCRP. Production of PAI-1 and endothelin-1 was little affected by either rhCRP preparation in either cell type. In addition, P-rhCRP dose-dependently diminished adrenomedullin release from both cell types, but did not affect adrenomedullin receptor expression or function. Our findings highlight the importance of removing endotoxin from commercial rCRP preparations and show that hCRP elicits atherogenic responses from HCASMCs, but not HCAECs. PMID- 14751239 TI - Polyol pathway-dependent osmotic and oxidative stresses in aldose reductase mediated apoptosis in human lens epithelial cells: role of AOP2. AB - Aldose reductase (AR) has been implicated as a major contributor to the pathogenesis of diabetic cataracts. AR activation generates osmotic and oxidative stresses via the polyol pathway and induces cell death signals. Antioxidant protein 2 (AOP2) protects cells from oxidative stress. We investigated the effect of AR overexpression on polyol accumulation and on hyperglycemic oxidative stress and osmotic stress, as well as the effects of these stresses on human lens epithelial cell (hLEC) survival. hLECs overexpressing the AR became apoptotic during hyperglycemia and showed elevated levels of intracellular polyols. Glutathione and AOP2 levels were significantly decreased in these cells. Interestingly, supply of AOP2 and/or the AR inhibitor fidarestat protected the cells against hyperglycemia-induced death. Overexpression of AR increased osmotic and oxidative stresses, resulting in increased apoptosis in hLECs. Because AOP2 protects hyperglycemia-induced hLEC apoptosis, this molecule may have the potential to prevent hyperglycemia-mediated complications in diabetes. PMID- 14751241 TI - Single intravenous injection of naked plasmid DNA encoding erythropoietin provides neuroprotection in hypoxia-ischemia rats. AB - Hypoxic-ischemic (H-I) encephalopathy is a major contributor to morbidity and mortality in infants and children. To delineate the nature and mechanism(s) of neuroprotection via erythropoietin (EPO) gene therapy, we evaluated the effects of single intravenous injection of naked plasmid DNA encoding EPO in H-I infant rats. Single administration of naked plasmid containing EPO cDNA driven under cytomegalovirus promoter (pCMV-EPO) by rapid injection via the tail vein produced a remarkable level of human EPO protein in the circulation, peaking at one day and lasting for 14 days after injection. There were significant improvements of water maze task in H-I rats after EPO gene therapy. Our data showed that the mechanisms of EPO gene therapy were rescue of CA1 neurons from lethal H-I injury, prevention of neuronal apoptosis in CA1 region, and decrease of glial activation in corpus callosum. This could be the first report of successful treatment of H-I injury by a single intravenous infusion of EPO gene. PMID- 14751242 TI - Interleukin-12 genetic administration suppressed metastatic liver tumor unsusceptible to CTL. AB - A cytokine gene therapy approach was conducted against metastatic lesions of cytotoxic T lymphocyte (CTL)-unsusceptible tumor in mice. The EBV-based and conventional plasmid vectors that encode murine interleukin-12 (IL-12) gene (pGEG.mIL-12 and pG.mIL-12, respectively) were intravenously transfected into the mice that had received a subcutaneous inoculation of M5076 sarcoma cells. The pGEG.mIL-12 transfection drastically suppressed the subcutaneous as well as hepatic metastatic tumors, resulting in significant prolongation of survival period of the animals. Although single administration with pG.mIL-12 was not effective, repetitive transfection with the plasmid significantly prolonged the longevity of the mice-bearing the metastatic liver tumors. Multiple transfection with either pGEG.mIL-12 or pG.mIL-12 also suppressed peritoneal carcinomatosis in mice that had been injected with M5076 cells into the peritoneal cavity. It was suggested that a high level IL-12 production elicited by the intravenous delivery of the cytokine gene may be quite effective in inhibiting metastatic and CTL unsusceptible neoplasms. PMID- 14751243 TI - Transient posttranslational up-regulation of telomerase activity during megakaryocytic differentiation of K562 cells. AB - Telomerase is active in immature somatic cells, but not in differentiated cells. However, the mechanism by which telomerase is regulated in relation to cell differentiation is not well understood. In this study, the human erythroid leukemia cell line K562 was induced to differentiate into megakaryocytes by TPA and into erythroid by STI571. The human acute myeloblastic leukemia cell line HL60 was also induced to differentiate into monocytes by TPA. Telomerase activity, the expression of human telomerase reverse transcriptase, hTERT, and the cell cycle were examined. TPA induced a transient increase in telomerase activity during the megakaryocytic differentiation while the message of hTERT decreased gradually throughout the same period. This suggests the existence of a regulatory mechanism other than transcription of hTERT. Cell cycle analysis revealed that cells in G(2)/M phase increased in number in accordance with the changes in telomerase activity. Pretreatment with PKC inhibitors inhibited the megakaryocytic differentiation, transient increase in telomerase activity, and G(2)/M arrest. These results suggest that PKC acts as a transient post translational activator of telomerase during megakaryocytic differentiation. PMID- 14751244 TI - Molecular cloning and expression of HRLRRP, a novel heart-restricted leucine-rich repeat protein. AB - We isolated a novel leucine-rich repeat protein (LRRP) cDNA from E13 mouse embryos by the in silico approach. The cDNA encoded a protein of 274 amino acids having 7 leucine-rich repeat motifs at the center of the protein. An in vitro transcription/translation study showed that the cDNA coded for a peptide of approximately 31kDa. Northern blot analysis suggested that the mRNA of this novel LRRP was expressed only in the heart, although RT-PCR indicated slight expression in skeletal muscle as well. The transcripts of this gene and Nkx-2.5/Csx were detected in the early stage of cardiac differentiation of P19CL6 embryonal carcinoma cells treated with 1% dimethyl sulfoxide. The fusion protein made between it and GFP was detected at a high level in mitochondria and a low level in the nuclei of COS7 cells. The nuclei of the adult mouse heart were strongly stained with the antibody raised against the synthetic peptide of the protein. Therefore, we designated the gene as heart-restricted leucine-rich repeat protein (HRLRRP) and assume that mouse HRLRRP may play important roles in cardiac development and/or cardiac function. PMID- 14751245 TI - Suppression of prostaglandin E2 receptor subtype EP2 by PPARgamma ligands inhibits human lung carcinoma cell growth. AB - Prostaglandin E(2) (PGE(2)), a major cyclooxygenase (COX-2) metabolite, plays important roles in tumor biology and its functions are mediated through one or more of its receptors EP1, EP2, EP3, and EP4. We have shown that the matrix glycoprotein fibronectin stimulates lung carcinoma cell proliferation via induction of COX-2 expression with subsequent PGE(2) protein biosynthesis. Ligands of peroxisome proliferator-activated receptor gamma (PPARgamma) inhibited this effect and induced cellular apoptosis. Here, we explore the role of the PGE(2) receptor EP2 in this process and whether the inhibition observed with PPARgamma ligands is related to effects on this receptor. We found that human non small cell lung carcinoma cell lines (H1838 and H2106) express EP2 receptors, and that the inhibition of cell growth by PPARgamma ligands (GW1929, PGJ2, ciglitazone, troglitazone, and rosiglitazone [also known as BRL49653]) was associated with a significant decrease in EP2 mRNA and protein levels. The inhibitory effects of BRL49653 and ciglitazone, but not PGJ2, were reversed by a specific PPARgamma antagonist GW9662, suggesting the involvement of PPARgamma dependent and -independent mechanisms. PPARgamma ligand treatment was associated with phosphorylation of extracellular regulated kinase (Erk), and inhibition of EP2 receptor expression by PPARgamma ligands was prevented by PD98095, an inhibitor of the MEK-1/Erk pathway. Butaprost, an EP2 agonist, like exogenous PGE(2) (dmPGE(2)), increased lung carcinoma cell growth, however, GW1929 and troglitazone blocked their effects. Our studies reveal a novel role for EP2 in mediating the proliferative effects of PGE(2) on lung carcinoma cells. PPARgamma ligands inhibit human lung carcinoma cell growth by decreasing the expression of EP2 receptors through Erk signaling and PPARgamma-dependent and -independent pathways. PMID- 14751246 TI - Conflicting effects of caffeine on apoptosis and clonogenic survival of human K1 thyroid carcinoma cell lines with different p53 status after exposure to cisplatin or UVc irradiation. AB - Caffeine has been widely described as a chemo/radiosensitizing agent, presumably by inhibiting DNA repair, and affecting preferentially cells with an altered p53 status. We evaluated the effects of caffeine using isogenic and isophenotypic K1 cells derived from a papillary thyroid carcinoma and displaying either a wild type or a mutated p53 status. Apoptosis and clonogenic survival were examined after exposure of the cells to cisplatin or UVc irradiation. We find that at the most currently used concentration, 2mM, caffeine hinders cisplatin or UVc induced apoptosis in K1 cells. In addition, at this already barely achievable concentration in vivo, caffeine does not decrease their clonogenic survival. Hence in our cellular model, caffeine does not behave as a chemo- or a radiosensitizer. Although surprising, these results (1) are in agreement with the delayed G2/M block caused by caffeine that we previously observed in normal human fibroblasts and K1 cells and (2) allow us to elucidate some discrepancies concerning this molecule throughout the literature such as increase or decrease of apoptosis and clonogenic survival, activation or deactivation of molecules involved in DNA damage repair and proliferation inhibition but accelerated G2/M traverse. PMID- 14751247 TI - Presence of interaction but not complementation between human mtDNAs carrying different mutations within a tRNA(Leu(UUR)) gene. AB - This study provided answers to fundamental questions on mammalian mitochondrial genetics: Could respiratory function in mitochondria be maintained by their exchange of genetic contents even when mutations were created within the same genes in different mitochondrial DNA (mtDNA) molecules? Using cell fusion techniques, we created a chance to coexist two types of respiration-deficient syn(-) mitochondria carrying different mtDNA mutations within the same tRNA(Leu(UUR)) gene obtained from patients with mitochondrial diseases. The results showed that two syn(-) mitochondria exchanged their genetic contents, but did not restore respiration defects, suggesting that mitochondrial interaction could not complement the mutations created within the same gene in different mtDNA molecules. PMID- 14751248 TI - Model analysis of difference between EGF pathway and FGF pathway. AB - The difference in time course of Ras and mitogen activated protein kinase (MAPK) cascade by different growth factors is considered to be the cause of different cellular responses. We have developed the computer simulation of Ras-MAPK signal transduction pathway containing newly identified negative feedback system, Sprouty, and adaptor molecules. Unexpectedly, negative feedback system did not profoundly affect time course of MAPK activation. We propose the key role of fibroblast growth factor receptor substrate 2 (FRS2) in NGF/FGF pathway for sustained MAPK activation. More Grb2-SOS complexes were recruited to the plasma membrane by binding to membrane-bound FRS2 in FGF pathway than in EGF pathway and caused sustained activation of ERK. The EGF pathway with high concentration of EGF receptor also induced sustained MAPK activation, which is consistent with the results in the PC12 cell overexpressing the EGF receptors. The simulated time courses of FRS2 knock-out cells were consistent with those of the reported experimental results. PMID- 14751249 TI - Expression of glucose transporter 4 in the human pancreatic islet of Langerhans. AB - Glucose transporter 4 (GLUT4) is the main insulin-responsive glucose transporter in skeletal muscle and adipose tissue of human and rodent, and is translocated to the plasma membrane in response to insulin. GLUT2 is well known as the main glucose transporter in pancreatic islets and could highly regulate glucose stimulated insulin secretion by B-cells as a glucose sensor. We confirmed the presence of GLUT4 mRNA and GLUT4 protein in pancreas in the human. Indirect immunohistochemistry showed that the pancreatic islets of human and rat were conspicuously labeled by anti-GLUT4 antibody. The presence of placental leucine aminopeptidase (P-LAP), a homologue of insulin-regulated aminopeptidase (IRAP), was also shown in the human pancreatic islet. IRAP/P-LAP is thought to be involved in glucose metabolism. This study provides the first evidence that GLUT4 is present in human and rat pancreatic islets and may suggest its specific role in glucose homeostasis in conjunction with IRAP/P-LAP. PMID- 14751250 TI - Differential regulation of host cellular genes by HIV-1 viral protein R (Vpr): cDNA microarray analysis using isogenic virus. AB - HIV-1 Vpr is a protein with multiple functions. It has been suggested that such pleiotropic effects by a viral protein may be mediated by its association with viral and cellular proteins or through modulation of expression of specific cellular genes. To address this, we used cDNA microarray techniques to analyze the regulation of a panel of host cellular genes by HIV-1 Vpr using isogenic HIV 1 either with or without Vpr expression. Results indicate that Vpr downregulated the expression of genes involved in cell cycle/proliferation regulation, DNA repair, tumor antigens, and immune activation factors, and upregulated many ribosomal and structural proteins. These results for the first time reveal the involvement of several potential cellular genes, which may be useful, both for understanding Vpr functions and for the development of therapeutics targeting the Vpr molecule. PMID- 14751251 TI - Clinico-molecular study of dedifferentiation in well-differentiated liposarcoma. AB - Well-differentiated liposarcoma (WD) acquires fully malignant potential when the histological progression named dedifferentiation occurs. This progression is supposed to occur in a time-dependent manner but this is still a debated issue. Clinically, the prediction of dedifferentiation for WD is very important from the therapeutic point of view. To identify genes that are predictive of dedifferentiation and to understand the mechanism of dedifferentiation, we investigated clinical information of 50 cases and studied the gene expression profiles of 36 lipomatous tumors using cDNA microarray. The clinical study showed that the dedifferentiation did not always seem to occur in a time-dependent manner. Interestingly, from the gene expression study, unsupervised hierarchical clustering analysis of well-differentiated lesions obtained from dedifferentiated liposarcoma (DD) cases that were indistinguishable from WD pathologically showed a clearly distinct gene expression pattern from WD. Using the pattern-matching program, 1687 genes including 487 known genes were identified, which discriminated WD cases from well-differentiated lipomatous lesions obtained from DD cases. These results suggest that the dedifferentiation may arise from different types of WD that could be distinguished from gene expression profiling but could hardly be classified by the pathological studies. PMID- 14751252 TI - Magnetic bead isolation of neutrophil plasma membranes and quantification of membrane-associated guanine nucleotide binding proteins. AB - A protocol for isolation of neutrophil plasma membranes utilizing a plasma membrane marker antibody, anti-CD15, attached to superparamagnetic beads was developed. Cells were initially disrupted by nitrogen cavitation and then incubated with anti-CD15 antibody-conjugated superparamagnetic beads. The beads were then washed to remove unbound cellular debris and cytosol. Recovered plasma membranes were quantified by immunodetection of G(beta2) in Western blots. This membrane marker-based separation yielded highly pure plasma membranes. This protocol has advantages over standard density sedimentation protocols for isolating plasma membrane in that it is faster and easily accommodates cell numbers as low as 10(6). These methods were coupled with immunodetection methods and an adenosine 5(')-diphosphate-ribosylation assay to measure the amount of membrane-associated G(ialpha) proteins available for receptor coupling in neutrophils either stimulated with N-formyl peptides or treated to differing degrees with pertussis toxin. As expected, pertussis toxin treatment decreased the amount of membrane G protein available for signaling although total membrane G protein was not affected. In addition, activation of neutrophils with N-formyl peptides resulted in an approximately 50% decrease in G protein associated with the plasma membrane. PMID- 14751253 TI - Cyclodextrin-bound 6-(4-methoxyphenyl)imidazo[1,2-alpha+/-]pyrazin-3(7H)-ones with fluorescein as green chemiluminescent probes for superoxide anions. AB - In providing chemiluminescent probes that have high chemiluminescence intensity and high specificity to superoxide anions, novel chemiluminescent probes involving cyclodextrins covalently bound to 6-(4-methoxyphenyl)imidazo[1,2 alpha]pyrazin-3(7H)-one with fluorescein were synthesized and characterized. Using the hypoxanthine-xanthine oxidase system for the generation of the superoxide anions, these novel chemiluminescent probes showed higher superoxide induced chemiluminescence intensity than that of 6-[4-[2-[N(')-(5 fluoresceinyl)thioureido]-ethoxy]phenyl]-2-methylimidazo[1,2-alpha]pyrazin-3(7H) one (FCLA). When tested at a probe concentration of 1.0 microM, compound 6, in which 6-(4-methoxyphenyl)imidazo[1,2-alpha]pyrazin-3(7H)-one and fluorescein are covalently attached on the secondary and primary hydroxyl faces of gamma cyclodextrin, respectively, showed green luminescence intensity that was 26 times that of FCLA, which was also the highest luminescence intensity in this present study. At probe concentrations of less than 1.0 microM, the ratio of the superoxide-dependent chemiluminescence intensity to the background chemiluminescence intensity for compound 6 was higher than that of FCLA. This high superoxide-induced chemiluminescence intensity and superoxide specificity in low probe concentrations indicates that 6 can be more effective than FCLA toward the measurement of superoxide anions. PMID- 14751254 TI - Analysis of medium-chain acyl-coenzyme A esters in mouse tissues by liquid chromatography-electrospray ionization mass spectrometry. AB - Medium-chain acyl-coenzyme A (CoA) esters are key metabolites in lipid metabolism. Liquid chromatography-electrospray ionization mass spectrometry analysis of medium-chain acyl-CoA esters is described. Eight medium-chain acyl CoA esters were well separated on a C(8)-MS reversed-phase column using a linear gradient of ammonium acetate buffer (pH 5.3)-acetonitrile. The positive-ion mass spectra of all the saturated and unsaturated medium-chain acyl-CoA esters gave dominant [M+H](+) ions, whereas their negative-ion mass spectra showed abundant [M-H](-) and [M-2H](2-) ions. The positive-ion mode of operation was slightly less sensitive than the negative-ion detection mode. Five medium-chain acyl-CoA esters of C(6:0), C(8:0), C(8:1), C(10:0), and C(10:1) in liver, heart, kidney, and brain from the mouse were identified. The predominant acyl-CoA peaks were C(6:0), C(8:0), and C(10:0). Small amounts of medium-chain acyl-CoAs of C(8:1) and C(10:1) were detected only in heart and kidney. The analytical method is very useful for the analysis of medium-chain acyl-CoA esters in the tissues. PMID- 14751255 TI - Determination of enantiomeric compositions of pharmaceutical products by near infrared spectrometry. AB - A new method for the determination of enantiomeric compositions of a variety of drugs including propranolol, atenolol, and ibuprofen has been developed. The method is based on the use of the near-infrared technique to measure diastereomeric interactions between an added carbohydrate compound and both enantiomeric forms of a drug followed by evaluation of the data by partial least square analysis. The fact that the method works well with all three macrocyclic carbohydrates with different cavity sizes (i.e., alpha-, beta-, and gamma cyclodextrin) and with sucrose, which is a linear carbohydrate, clearly demonstrates that it is not necessary to have inclusion complex formation to produce effective diastereomeric interactions. Rather a simple adsorption of the drug onto a carbohydrate is sufficient. Since inclusion complex formation is not a requisite, this method is not limited to the three drugs evaluated in this study but is rather universal as it can, in principle, be used for the sensitive and accurate determination of enantiomeric compositions of many different types of drugs with only about 1.5mg/mL concentration and enantiomeric excess as low as 0.80%, in water or in a mixture of water with organic solvent. Furthermore, it does not rely on the use of rather expensive carbohydrates such as cyclodextrins but is equally as effective even with a simple and inexpensive carbohydrate such as sucrose. PMID- 14751256 TI - Chaotic mixer improves microarray hybridization. AB - Hybridization is an important aspect of microarray experimental design which influences array signal levels and the repeatability of data within an array and across different arrays. Current methods typically require 24h and use target inefficiently. In these studies, we compare hybridization signals obtained in conventional static hybridization, which depends on diffusional target delivery, with signals obtained in a dynamic hybridization chamber, which employs a fluid mixer based on chaotic advection theory to deliver targets across a conventional glass slide array. Microarrays were printed with a pattern of 102 identical probe spots containing a 65-mer oligonucleotide capture probe. Hybridization of a 725 bp fluorescently labeled target was used to measure average target hybridization levels, local signal-to-noise ratios, and array hybridization uniformity. Dynamic hybridization for 1h with 1 or 10ng of target DNA increased hybridization signal intensities approximately threefold over a 24-h static hybridization. Similarly, a 10- or 60-min dynamic hybridization of 10ng of target DNA increased hybridization signal intensities fourfold over a 24h static hybridization. In time course studies, static hybridization reached a maximum within 8 to 12h using either 1 or 10ng of target. In time course studies using the dynamic hybridization chamber, hybridization using 1ng of target increased to a maximum at 4h and that using 10ng of target did not vary over the time points tested. In comparison to static hybridization, dynamic hybridization reduced the signal-to noise ratios threefold and reduced spot-to-spot variation twofold. Therefore, we conclude that dynamic hybridization based on a chaotic mixer design improves both the speed of hybridization and the maximum level of hybridization while increasing signal-to-noise ratios and reducing spot-to-spot variation. PMID- 14751257 TI - Characterization and analysis of thermal denaturation of antibodies by size exclusion high-performance liquid chromatography with quadruple detection. AB - Size exclusion chromatography (SEC) coupled with online light scattering, viscometry, refractometry, and UV-visible spectroscopy provides a very powerful tool for studying protein size, shape, and aggregation. This technique can be used to determine the molecular weight of the component peaks independent of the retention times in the SEC column and simultaneously measure the hydrodynamic radius and polydispersity of the protein. We applied this technology by coupling an Agilent Chemstation high-performance liquid chromatography system with a diode array UV-visible detector and a Viscotek 300 EZ Pro triple detector (combination of a light scattering detector, refractometer, and differential pressure viscometer) to characterize and compare the molecular properties of a number of monoclonal antibodies. Our studies reveal that different monoclonal immunoglobulin Gs (IgGs) and chimeric IgGs show slightly different retention times and therefore different molecular weights in gel filtration analysis. However, when they are analyzed by light scattering, refractometry, and viscometry, different IgGs have comparable molecular weight, molecular homogeneity (polydispersity), and size. Gel filtration coupled with UV or refractive index detection suggests that antibodies purified and formulated for preclinical and clinical development are more than 95% monomer with little or no detectable soluble aggregates. Light scattering measurements showed the presence of trace amounts of soluble aggregate in all the IgG preparations. The different IgG molecules showed different susceptibility to heat and pH. One of the murine antibodies was considerably less stable than the others at 55 degrees C. The application of this powerful technology for the characterization of monoclonal antibodies of therapeutic potential is discussed. PMID- 14751258 TI - Bioluminescent immunoassay of thyrotropin and thyroxine using obelin as a label. AB - Solid-phase bioluminescent immunoassay of thyroid hormones, human thyrotropin (hTSH) and two forms of thyroxine (T4), whose determinations are vitally important for diagnostics of thyroid diseases and the efficiency of treatment, is described. The recombinant obelin, a Ca(2+)-regulated photoprotein originally derived from the luminous marine hydroid Obelia longissima, is employed as a bioluminescent label. To produce obelin conjugates with anti-hTSH, anti-T4 immunoglobulins (IgG), and T4, additional SH groups are introduced into the obelin molecule using Traut's reagent (2-iminothiolane) and then obelin possessing extra SH groups is conjugated with succinimidyl 4-(N-maleimidomethyl) cyclohexane-1-carboxylate-activated IgGs or T4. The total yield of obelin conjugates determined by luminescent activity is 60-65% after all chemical and purification procedures. The obtained conjugates are stable to lyophilization and in solution for at least 9 months at 4 degrees C, with loss of activity not exceeding 10%. The application of obelin conjugates for determination of the hTSH, total T4, and free T4 in standard, control, and patient sera displays high sensitivity and reproducibility of results. The results of bioluminescent immunoassays are closely comparable to those obtained by the radioimmunoassay method (R=0.95-0.99). PMID- 14751259 TI - A continuous nonradioactive assay for RNA-dependent RNA polymerase activity. AB - Current assays for the activity of viral RNA-dependent RNA polymerases (RdRps) are inherently end-point measurements, often requiring the use of radiolabeled or chemically modified nucleotides to detect reaction products. In an effort to improve the characterization of polymerases that are essential to the life cycle of RNA viruses and develop antiviral therapies that target these enzymes, a continuous nonradioactive assay was developed to monitor the activity of RdRps by measuring the release of pyrophosphate (PP(i)) generated during nascent strand synthesis. A coupled-enzyme assay method based on the chemiluminescent detection of PP(i), using ATP sulfurylase and firefly luciferase, was adapted to monitor poliovirus 3D polymerase (3D(pol)) and the hepatitis C virus nonstructural protein 5B (NS5B) RdRp reactions. Light production was dependent on RdRp and sensitive to the concentration of oligonucleotide primer directing RNA synthesis. The assay system was found to be amenable to sensitive kinetic studies of RdRps, requiring only 6nM 3D(pol) to obtain a reliable estimate of the initial velocity in as little as 4 min. The assay can immediately accommodate the use of both homopolymer and heteropolymer RNA templates lacking uridylates and can be adapted to RNA templates containing uridine by substituting alpha-thio ATP for ATP. The low background signal produced by other NTPs can be corrected from no enzyme (RdRp) controls. The effect of RdRp/RNA template preincubation was assessed using NS5B and a homopolymer RNA template and a time-dependent increase of RdRp activity was observed. Progress curves for a chain terminator (3(') deoxyguanosine 5(')-triphosphate) and an allosteric NS5B inhibitor demonstrated the predicted time- and dose-dependent reductions in signal. This assay should facilitate detailed kinetic studies of RdRps and their potential inhibitors using either standard or single-nucleotide approaches. PMID- 14751260 TI - Monitoring the formation of Maillard reaction products of glucosamine with fibrinogen and human serum albumin using capillary electrophoresis. AB - Formation of Maillard reaction products (MRP) of glucosamine (GlcN) with fibrinogen and human serum albumin (HSA), under simulated physiological conditions, was detected by fluorescence (excitation/emission: 340/420 nm) and UV/Vis (max. 275 nm) spectroscopy. The application of polyacrylamide gel electrophoresis demonstrated the generation of high-molecular-weight fibrinogen and HSA MRP by GlcN. A simple and rapid capillary electrophoresis method was developed to separate MRP formed by the reaction of GlcN with proteins from GlcN autocondensation products. PMID- 14751261 TI - Use of relaxation-edited one-dimensional and two dimensional nuclear magnetic resonance spectroscopy to improve detection of small metabolites in blood plasma. AB - The 1H nuclear magnetic resonance (NMR) spectra of biological samples, such as blood plasma and tissues, are information rich but data complex owing to superposition of the resonances from a multitude of different chemical entities in multiple-phase compartments, hampering detection and subsequent resonance assignments. To overcome these problems, several spectral-editing NMR experiments are described here, combining spin-relaxation filters (based on T(1), T(rho), and T(2)) with both one-dimensional and two-dimensional (2D) NMR spectroscopy. These techniques enable the separation of NMR resonances based on their relaxation times and allow simplification of the complex spectra. In this paper, the approach is exemplified using a control human blood plasma, which is a complex mixture of proteins, lipoproteins, and small-molecule metabolites. In the case of T(1rho)- and T(2)-edited 2D NMR experiments, a "flip-back" pulse was introduced after the relaxation editing to make the phase cycling of the "relaxation filter" and the 2D NMR part independent, thus enabling easy implementation of the phase sensitive 2D NMR experiments. These methods also permit much higher receiver gains to be used to reduce digitization error, in particular, for the small resonances, which are sometimes vitally important for metabonomics studies. Both pulse sequences and experimental results are discussed for T(1)-, T(1rho)-, and T(2)-filtered COSY, T(2)-filtered phase-sensitive DQF-COSY, and T(1), T(1rho)-, and T(2)-filtered TOCSY NMR. PMID- 14751262 TI - Fluorescence labeling, purification, and immobilization of a double cysteine mutant calmodulin fusion protein for single-molecule experiments. AB - We present a method of labeling and immobilizing a low-molecular-weight protein, calmodulin (CaM), by fusion to a larger protein, maltose binding protein (MBP), for single-molecule fluorescence experiments. Immobilization in an agarose gel matrix eliminates potential interactions of the protein and the fluorophore(s) with a glass surface and allows prolonged monitoring of protein dynamics. The small size of CaM hinders its immobilization in low-weight-percentage agarose gels; however, fusion of CaM to MBP via a flexible linker provides sufficient restriction of translational mobility in 1% agarose gels. Cysteine residues were engineered into MBP.CaM (MBP-T34C,T110C-CaM) and labeled with donor and acceptor fluorescent probes yielding a construct (MBP.CaM-DA) which can be used for single molecule single-pair fluorescence resonance energy transfer (spFRET) experiments. Mass spectrometry was used to verify the mass of MBP.CaM-DA. Assays measuring the activity of CaM reveal minimal activity differences between wild-type CaM and MBP.CaM-DA. Single-molecule fluorescence images of the donor and acceptor dyes were fit to a two-dimensional Gaussian function to demonstrate colocalization of donor and acceptor dyes. FRET is demonstrated both in bulk fluorescence spectra and in fluorescence trajectories of single MBP.CaM-DA molecules. The extension of this method to other biomolecules is also proposed. PMID- 14751263 TI - Direct electron transfer and bioelectrocatalysis of hemoglobin at a carbon nanotube electrode. AB - A stable suspension of carbon nanotube (CNT) can be obtained by dispersing the CNT in the solution of the surfactant cetyltrimethylammonium bromide. CNT has promotion effects on the direct electron transfer of hemoglobin (Hb), which was immobilized onto the surface of CNT. The direct electron transfer rate of Hb was greatly enhanced after it was immobilized onto the surface of CNT. Cyclic voltammetric results showed a pair of well-defined redox peaks, which corresponded to the direct electron transfer of Hb, with the formal potential (E(0('))) at about -0.343V (vs. saturated calomel electrode) in the phosphate buffer solution (pH 6.8). The electrochemical parameters such as apparent heterogeneous electron transfer rate constant (k(s)) and the value of formal potential (E(0('))) were estimated. The dependence of E(0(')) on solution pH indicated that the direct electron transfer reaction of Hb is a one-electron transfer coupled with a one-proton transfer reaction process. The experimental results also demonstrated that the immobilized Hb retained its bioelectrocatalytic activity to the reduction of H(2)O(2). The electrocatalytic current was proportional to the concentration of H(2)O(2) at least up to 20mM. PMID- 14751264 TI - Atomic force microscope investigation of large-circle DNA molecules. AB - A circular bacterial artificial chromosome of 148.9kbp on human chromosome 3 has been extended and fixed on bare mica substrates using a developed fluid capillary flow method in evaporating liquid drops. Extended circular DNA molecules were imaged with an atomic force microscope (AFM) under ambient conditions. The measured total lengths of the whole DNA molecules were in agreement with sequencing analysis data with an error range of +/-3.6%. This work is important groundwork for probing single nucleotide polymorphisms in the human genome, mapping genomic DNA, manipulating biomolecular nanotechnology, and studying the interaction of DNA-protein complexes investigated by AFM. PMID- 14751265 TI - Kinetic screening of antibodies from crude hybridoma samples using Biacore. AB - Experimental and data analysis protocols were developed to screen antibodies from hybridoma culture supernatants using Biacore surface plasmon resonance biosensor platforms. The screening methods involved capturing antibodies from crude supernatants using Fc-specific antibody surfaces and monitoring antigen binding at a single concentration. After normalizing the antigen responses for the amount of antibody present, a simple interaction model was fit to all of the binding responses simultaneously. As a result, the kinetic rate constants (k(a) and k(d)) and affinity (K(D)) could be determined for each antibody interaction under identical conditions. Higher-resolution studies involving multiple concentrations of antigen were performed to validate the reliability of single-concentration measurements. The screening protocols can be used to characterize antigen binding kinetics to approximately 200 antibody supernatants per day using automated Biacore 2000 and 3000 instruments. PMID- 14751266 TI - High-throughput metabolic flux analysis based on gas chromatography-mass spectrometry derived 13C constraints. AB - 13C-constrained flux balancing analysis based on gas chromatography-mass spectrometry data is presented here as a simple and robust method for the estimation of intracellular carbon fluxes. In this approach, the underdetermined system of metabolite balances deduced from stoichiometric relations and measured extracellular rates is complemented with 13C constraints from metabolic flux ratio analysis. Fluxes in central carbon metabolism of exponentially growing Escherichia coli were estimated by 13C-constrained flux balancing from three different 13C-labeled glucose experiments. The best resolution of the network was achieved using 13C constraints derived from [U-13C]glucose and [1-13C]glucose experiments. The corresponding flux estimate was in excellent agreement with a solution that was independently obtained with a comprehensive isotopomer model. This new methodology was also demonstrated to faithfully capture the intracellular flux distribution in E. coli shake flasks and 1-ml deep-well microtiter plates. Due to its simplicity, speed, and robustness, 13C-constrained metabolic flux balancing is promising for routine and high-throughput analysis on a miniaturized scale. PMID- 14751267 TI - Caspase multiplexing: simultaneous homogeneous time-resolved quenching assay (TruPoint) for caspases 1, 3, and 6. AB - Caspases are a group of cysteine proteases involved in apoptosis and inflammation. A multiparametric homogeneous assay capable of measuring activity of three different caspases in a single well of a microtiter plate is described. Different fluorescent europium, samarium, terbium, and dysprosium chelates were coupled to a caspase substrate peptide, their luminescence properties, were analyzed, and their function in a time-resolved fluorescence quenching-based caspase 3 assay was studied. Substrates for caspases 1, 2, 3, 6, and 8 and granzyme B were also synthesized and their specificities for different caspases were determined. By selecting suitable lanthanide chelates and substrates we developed a multiparametric homogeneous time-resolved fluorescence quenching based assay for caspases 1, 3, and 6. The assay was capable of measuring the activity of both single caspases and a mixture of three caspases mixed in the same well. PMID- 14751268 TI - Direct observation of covalent adducts with Cys34 of human serum albumin using mass spectrometry. AB - The interactions of the unpaired thiol residue (Cys34) of human serum albumin (HSA) with low-molecular-weight thiols and an Au(I)-based antiarthritic drug have been examined using electrospray ionization mass spectrometry. Early measurements of the amount of HSA containing Cys34 as the free thiol suggested that up to 30% of circulating HSA bound cysteine as a mixed disulfide. It has also been suggested that reaction of HSA with cysteine, occurs only on handling and storage of plasma. In our experiments, there were three components of HSA in freshly collected plasma from normal volunteers, HSA, HSA+cysteine, and HSA+glucose in the ratio approximately 50:25:25. We addressed this controversy by using iodoacetamide to block the free thiol of HSA in fresh plasma, preventing its reaction with plasma cysteine. When iodoacetamide was injected into a vacutaner tube as blood was collected, the HSA was modified by iodoacetamide, with 20-30% present as the mixed disulfide with cysteine (HSA+cys). These data provide strong evidence that 20-30% of HSA in normal plasma contains one bound cysteine. Reaction of HSA with [Au(S(2)O(3))(2)](3-) resulted in formation of the adducts HSA+Au(S(2)O(3)) and HSA+Au. Reaction of HSA with iodoacetamide prior to treatment with [Au(S(2)O(3))(2)](3-) blocked the formation of gold adducts. PMID- 14751269 TI - A fluorimetric method using fluorescein di-beta-D-galactopyranoside for quantifying the senescence-associated beta-galactosidase activity in human foreskin fibroblast Hs68 cells. AB - The senescence-associated beta-galactosidase (SA-betaG) assay is one of the few accepted markers of cell aging. However, the cytochemical method using 5-bromo-4 chloro-3-indolyl beta-D-galactopyranoside (X-Gal) as substrate is limited in sensitivity and is only semiquantitative. Here, we modified the X-Gal method by replacing X-Gal with fluorescein di-beta-D-galactopyranoside (FDG) as substrate for SA-betaG, and the activity was measured fluorimetrically. We showed in Hs68 cells that the FDG fluorescein fluorescence increased with increasing passages of the cells in parallel with the X-Gal method. A major advantage of the FDG method is that it is a quantitative method for the SA-betaG activity. For example, we showed that the FDG fluorescein in p30(+1) of Hs68 cells was generally stronger than that in p26(+1) cells, whereas the X-Gal method gave similar results (95 and 100%) for p26(+1) and p30(+1) cells. The FDG method was precise with a relative standard deviation lower than 10%. We further demonstrated that FDG and X-Gal could be added simultaneously for SA-betaG assay because the FDG fluorescein diffused readily through formaldehyde-fixed cell membrane and could be detected in the suspension buffer. Thus, a double-substrate method, i.e., X-Gal for rapid qualitative assay and FDG for quantitative assay, can be conducted simultaneously to provide a simple and reliable assay of SA-betaG activity as a marker of cell aging. PMID- 14751270 TI - Studies of lipid turnover in cells with stable isotope and gas chromatograph-mass spectrometry. AB - Phospholipids are major building blocks for biological membranes. In addition, metabolites derived from their degradation are important signals in major cellular events, such as proliferation and apoptosis. The concept of lipid signaling in cells is derived mainly from the measurement of change in the concentration of lipid molecules. However, these changes in concentration are only a small part of the underlying metabolic change induced by a perturbation in the cell. In contrast, metabolic kinetic studies documenting product-precursor relationships and turnover rates are useful in elucidating the responsible mechanisms. Historically, metabolic studies of phospholipids in cells have been carried out with pulse or pulse-chase methods using radioactive isotopes. While these studies provide valuable information, their scope is restricted by inherent limitations. In this paper we describe a method using [1,2,3,4-13C(4)]palmitate as the tracer for studying the metabolic kinetics of the molecular species of diacylglycerol, ceramide, phosphatidylcholine, and sphingomyelin. After growing cells in the presence of labeled palmitate complexed to serum albumin, the lipids are extracted and separated into lipid classes. After enzymatic hydrolysis, diacylglyerols and ceramides as bis-trimethylsilyl derivatives are determined quantitatively with capillary column gas chromatography. Internal standards for each lipid class are used in the procedure. In addition, the isotopic enrichments of the lipid molecular species are determined with gas chromatograph-mass spectrometry. We applied this method to the study of HL60 cells. Different turnover rates were found for various molecular species. In addition, the sn-1 and sn-2 acyl groups appear to be synthesized at different rates for different molecular species. Other information, such as chain elongation and desaturation, might also be derived through the use of this method. PMID- 14751271 TI - A gas chromatography-mass spectrometry method for the measurement of fatty acid omega and omega(-1) hydroxylation kinetics by CYP4A1 using an artificial membrane system. AB - A gas chromatography-mass spectrometry assay method for the analysis of lauric, myristic, and palmitic acids and their omega and omega(-1) hydroxylated metabolites from in vitro incubations of cytochrome P450 CYP4A1, involving solid phase extraction and trimethysilyl derivatization, was developed. The assay was linear, precise, and accurate over the range 0.5 to 50microM for all the analytes. It has the advantages of a more rapid analysis time, an improved sensitivity, and a wider range of analytes compared with other methods. An artificial membrane system was optimized for application to purified CYP4A1 enzyme by investigating the molar ratios of cytochrome b(5) and cytochrome P450 reductase present in the incubation mixture. Using this method, the kinetics of omega and omega(-1) oxidation of lauric, myristic, and palmitic acids by CYP4A enzymes were measured and compared in rat liver microsomes and an artificial membrane system. PMID- 14751272 TI - A microtiter plate assay for assessing the interaction of eukaryotic initiation factor eIF4E with eIF4G and eIF4E binding protein-1. AB - Modulation of interactions among proteins is an important mechanism for regulating both the subcellular location and the function of proteins. An example of the importance of protein-protein interaction is the reversible association of eukaryotic initiation factor eIF4E with the eIF4E binding proteins 4E-BP1 and eIF4G. When bound to 4E-BP1, eIF4E cannot bind to eIF4G to form the active eIF4F complex, an event that is required for the binding of mRNA to the ribosome. Thus, association of eIF4E with 4E-BP1 represses mRNA translation by preventing the binding of mRNA to the ribosome. Previous studies have measured the amount of 4E BP1 or eIF4G bound to eIF4E by either affinity chromatography or immunoprecipitation of eIF4E followed by Western blot analysis for quantitation of 4E-BP1 and eIF4G. Both of these techniques have significant limitations. In the present study, we describe a microtiter plate-based assay for quantitation of the amount of 4E-BP1 and eIF4G bound to eIF4E that obviates many of the limitations of the earlier approaches. It also has the advantage that absolute amounts of the individual proteins can be easily estimated. The approach should be applicable to the study of a wide variety of protein-protein interactions. PMID- 14751274 TI - Up-regulation of gamma-aminobutyric acid transporter I mediates ethanol sensitivity in mice. AB - Ethanol is among the most widely abused drugs in the world. Chronic ethanol consumption leads to ethanol tolerance and addiction, and impairs learning and memory. Na+/Cl- dependent GABA transporters play an important role in controlling the concentration of GABA in the synaptic cleft, and thus they control the intensity and duration of synaptic transmission of GABA. It has been suggested that GABAergic system is involved in ethanol consumption, tolerance and addiction, because chronic ethanol consumption alters the expression of GABAA receptors and drugs on GABA receptors affect ethanol actions. The results of the present study reveal that that activity of GABA transporters in mouse brain after 15-min acute ethanol injection or after chronic ethanol consumption is increased. Moreover, mice pre-injected with a competitive or a noncompetitive antagonist of gamma-aminobutyric acid transporter subtype 1 (GAT1) showed high sensitivity to the sedative/hypnotic effects of ethanol. In contrast, transgenic mice overexpressing GAT1 displayed low sensitivity to ethanol, as shown by the righting reflex test. Mice overexpressing GAT1 survived a lethal dose of ethanol (9 g/kg, i.p.) longer, maintained locomotor activity longer after a sub-lethal dose (1.75 g/kg, i.p.) and exhibited a higher median lethal dose than wild-type littermates. These results suggest that GAT1 plays an important role in sensitivity to ethanol, and might be a therapeutic target for alcoholism prevention and treatment. Acute and chronic ethanol administration resulted in the increase of GABA transporter function. Use of GAT1 selective inhibitors and GAT1 overexpressing mice thus demonstrate that GAT1 should be an important protein mediating sensitivity to ethanol in mice. PMID- 14751275 TI - Sex differences in the effect of ethanol injection and consumption on brain allopregnanolone levels in C57BL/6 mice. AB - The pharmacological profile of allopregnanolone, a neuroactive steroid that is a potent positive modulator of gamma-aminobutyric acidA (GABAA) receptors, is similar to that of ethanol. Recent findings indicate that acute injection of ethanol increased endogenous allopregnanolone to pharmacologically relevant concentrations in male rats. However, there are no comparable data in mice, nor has the effect of ethanol drinking on endogenous allopregnanolone levels been investigated. Therefore, the present studies measured the effect of ethanol drinking and injection on allopregnanolone levels in male and female C57BL/6 mice. One group was given 17 days of 2-h limited access to a 10% v/v ethanol solution in a preference-drinking paradigm, while another group had access to water only. The ethanol dose consumed in 2 h exceeded 2 g/kg. Then, separate groups of mice were injected with either 2 g/kg ethanol or saline. Mice were killed 30 min after the 2-h drinking session or injection. Blood ethanol concentration was significantly higher in the ethanol-injected versus ethanol drinking groups, even though the dose was similar. Consumption of ethanol significantly increased brain allopregnanolone levels in male but not female mice, compared with animals drinking water, but did not alter plasma corticosterone levels. In contrast, injection of ethanol did not significantly alter brain allopregnanolone levels in male or female mice and only significantly increased plasma corticosterone levels in the male mice, when compared with saline-injected animals. The sex differences in the effect of ethanol administration on endogenous allopregnanolone levels suggest that the hormonal milieu may impact ethanol's effect on GABAergic neurosteroids. Importantly, these data are the first to report the effect of ethanol drinking on allopregnanolone levels and indicate that ethanol consumption and ethanol injection can produce physiologically relevant allopregnanolone levels in male mice. These results have important implications for studies investigating the potential role of endogenous allopregnanolone levels in modulating susceptibility to ethanol abuse. PMID- 14751276 TI - An animal model with relevance to schizophrenia: sex-dependent cognitive deficits in osteogenic disorder-Shionogi rats induced by glutathione synthesis and dopamine uptake inhibition during development. AB - Low glutathione levels have been observed in the prefrontal cortex and the cerebrospinal fluid of schizophrenic patients, possibly enhancing the cerebral susceptibility to oxidative stress. We used osteogenic disorder Shionogi mutant rats, which constitute an adequate model of the human redox regulation because both are unable to synthesize ascorbic acid. To study the long-term consequences of a glutathione deficit, we treated developing rats with L-buthionine-(S,R) sulfoximine (BSO), an inhibitor of glutathione synthesis, and later investigated their behavior until adulthood. Moreover, some rats were treated with the dopamine uptake inhibitor GBR 12909 in order to elevate dopamine extracellular levels, thereby mimicking the dopamine hyperactivity proposed to be involved in schizophrenia. BSO and GBR 12909 alone or in combination minimally affected the development of spontaneous alternation or basic sensory and motor skills. A major effect of BSO alone or in combination with GBR 12909 was the induction of cataracts in both sexes, whereas GBR 12909 induced an elevation of body weight in females only. Sex and age-dependent effects of the treatments were observed in a test of object recognition. At postnatal day 65, whereas male rats treated with both BSO and GBR 12909 failed to discriminate between familiar and novel objects, females were not affected. At postnatal day 94, male object recognition capacity was diminished by BSO and GBR 12909 alone or in combination, whereas females were only affected by the combination of both drugs. Inhibition of brain glutathione synthesis and dopamine uptake in developing rats induce long-term cognitive deficits occurring in adulthood. Males are affected earlier and more intensively than females, at least concerning object recognition. The present study suggests that the low glutathione levels observed in schizophrenic patients may participate in the development of some of their cognitive deficits. PMID- 14751277 TI - Serotonergic response to social stress and artificial social sign stimuli during paired interactions between male Anolis carolinensis. AB - Serotonergic activity is influenced by social status and manipulation of social signals. In the lizard Anolis carolinensis, eyespot formation, i.e. darkening of postorbital skin from green to black, appears during stressful and agonistic situations, forming first in males that become dominant. To assess the effect of eyespots on central serotonergic activity during social interaction, males were paired by weight and painted postorbitally with green or black paint. Manipulation of eyespot color influenced social interactions and status. All males that viewed an opponent with black painted eyespots became subordinate. In these subordinate animals, serotonergic activity was elevated in hippocampus, striatum, nucleus accumbens and locus ceruleus. In contrast, males that viewed opponents with hidden eyespots (painted green) and became dominant had increased serotonergic activity in hypothalamus, medial amygdala and raphe. Pre-painted eyespots produced results that distinguish dominant and subordinate relationships based on serotonergic activity not previously seen in unmanipulated pairs. Results from experiments using pairs are similar to those using mirrors for medial amygdala and locus ceruleus, but not hippocampus, nucleus accumbens or raphe. Decreased hypothalamic serotonin was associated with increased aggressive behavior. These results, when compared with previous studies, suggest some flexibility in central serotonergic systems, which may shape dominant and subordinate rank acquisition, and appear to be influenced by the completion of social role formation. Furthermore, social status and central serotonergic activity was influenced by a visual cue, the presence or absence of postorbital eyespots on an opponent. PMID- 14751278 TI - Early stages of memory formation in filial imprinting: Fos-like immunoreactivity and behavior in the domestic chick. AB - Early stages of memory formation in filial imprinting were studied in domestic chicks. Chicks trained for 15 min showed strong imprinting, demonstrated by a strong preference for their training stimulus, and the time course of this preference over 2 days after training was similar to that of chicks trained for 60 min. The chicks therefore learned characteristics of the training stimulus very early during training. The intermediate and medial hyperstriatum ventrale (IMHV) is a part of the chick forebrain that is crucial for imprinting. Previous experiments have shown a learning-specific increase in Fos-like immunoreactivity, used as a marker of neuronal activity, in the IMHV after training for 60 min. The time course of Fos expression in the IMHV was measured after training for 15 min and 60 min. The same pattern of expression was found for both training times, showing a peak 120 min after the start of training. The time course of expression was stimulus-dependent. Fos expression in the IMHV, but not the hippocampus, was significantly correlated with strength of imprinting. It is concluded that the learning-specific change in Fos expression in the IMHV is associated with very early components of memory formation. PMID- 14751279 TI - Long-term behavioral and neuronal cross-sensitization to amphetamine induced by repeated brief social defeat stress: Fos in the ventral tegmental area and amygdala. AB - Repeated exposure to stress induces cross-sensitization to psychostimulants. The present study assessed functional neural activation during social defeat stress induced sensitization to a subsequent amphetamine challenge. Social defeat stress was induced in intruder rats during short confrontations with an aggressive resident rat once every third day during the course of 10 days. Rats received d amphetamine injections (1 mg/kg, i.p.) 17 or 70 days after the first social defeat stress exposure. Amphetamine administration induced a significantly higher frequency of locomotor activity in stressed animals than in handled control rats, which was still evident 2 months after the last social stress exposure. Immunohistochemistry for Fos-like proteins was used to detect activated neural profiles in the striatum, nucleus accumbens (NAc), prefrontal cortex, amygdala, and ventral tegmental area (VTA). Repeated social defeat stress significantly increased Fos-like immunoreactive (Fos-LI) labeling 17 days after the start of stress exposure in the prelimbic and infralimbic cortical regions, NAc shell and core, medial, central and basolateral amygdala, and VTA, which probably represented the expression of chronic Fos-related antigens. Amphetamine augmented stress-induced Fos-LI labeling 17 days after the first stress episode in the dorsal striatum, NAc core, and medial amygdala, reflecting a cross-sensitization of Fos response. Amphetamine challenge 70 days after social stress exposures revealed sensitized Fos-LI labeling in the VTA and the amygdala. These data suggest that episodes of repeated social stress induce a long-lasting neural change that leads to an augmented functional activation in the VTA and amygdala, which might represent a neurobiological substrate for long-lasting cross sensitization of repeated social defeat stress with psychostimulant drugs. PMID- 14751280 TI - Altered expression of potassium channel subunit mRNA and alpha-dendrotoxin sensitivity of potassium currents in rat dorsal root ganglion neurons after axotomy. AB - Previous studies have raised the possibility that a decrease in voltage-gated K+ currents may contribute to hyperexcitability of injured dorsal root ganglion (DRG) neurons and the emergence of neuropathic pain. We examined the effects of axotomy on mRNA levels for various Kv1 family subunits and voltage-gated K+ currents in L4-L5 DRG neurons from sham-operated and sciatic nerve-transected rats. RNase protection assay revealed that Kv1.1 and Kv 1.2 mRNAs are highly abundant while Kv1.3, Kv1.4, Kv1.5 and Kv1.6 mRNAs were detected at lower levels in L4-L5 DRGs from sham and intact rats. Axotomy significantly decreased Kv1.1, Kv1.2, Kv1.3 and Kv1.4 mRNA levels by approximately 35%, approximately 60%, approximately 40% and approximately 80%, respectively, but did not significantly change Kv1.5 or Kv1.6 mRNA levels. Patch clamp recordings revealed two types of K+ currents in small-sized L4-L5 DRG neurons: sustained delayed rectifier currents elicited from a -40 mV holding potential and slowly inactivating A-type currents that was additionally activated from a -120 mV holding potential. Axotomy decreased both types of K+ currents by 50-60% in injured DRG neurons. In addition, axotomy increased the alpha-dendrotoxin sensitivity of the delayed rectifier, but not slow A-type K+ currents in injured DRG neurons. These results suggest that Kv1.1 and Kv1.2 subunits are major components of voltage-gated K+ channels in L4-L5 DRG neurons and that the decreased expression of Kv1-family subunits significantly contributes to the reduction and altered kinetics of Kv current in axotomized neurons. PMID- 14751281 TI - Initiation of electrographic seizures by neuronal networks in entorhinal and perirhinal cortices in vitro. AB - The hippocampus is often considered to play a major role in the pathophysiology of mesial temporal lobe epilepsy. However, emerging clinical and experimental evidence suggests that parahippocampal areas may contribute to a greater extent to limbic seizure initiation, and perhaps epileptogenesis. To date, little is known about the participation of entorhinal and perirhinal networks to epileptiform synchronization. Here, we addressed this issue by using simultaneous field potential recordings in horizontal rat brain slices containing interconnected limbic structures that included the hippocampus proper. Epileptiform discharges were disclosed by bath applying the convulsant drug 4 aminopyridine (50 microM) or by superfusing Mg(2+)-free medium. In the presence of 4-aminopyridine, slow interictal- (duration=2.34+/-0.29 s; interval of occurrence=25.75+/-2.11 s, n=16) and ictal-like (duration=31.25+/-3.34 s; interval of occurrence=196.96+/-21.56 s, n=17) discharges were recorded in entorhinal and perirhinal cortices after abating the propagation of CA3-driven interictal activity to these areas following extended hippocampal knife cuts. Simultaneous recordings obtained from the medial and lateral entorhinal cortex, and from the perirhinal cortex revealed that interictal and ictal discharges could initiate from any of these areas and propagate to the neighboring structure with delays of 8-66 ms. However, slow interictal- and ictal-like events more often originated in the medial entorhinal cortex and perirhinal cortex, respectively. Cutting the connections between entorhinal and perirhinal cortices (n=10), or functional inactivation of cortical areas by local application of a glutamatergic receptor antagonist (n=11) made independent epileptiform activity occur in all areas. These procedures also shortened ictal discharge duration in the entorhinal cortices, but not in the perirhinal area. Similar results could be obtained by applying Mg(2+)-free medium (n=7). These findings indicate that parahippocampal networks provide independent epileptiform synchronization sufficient to sustain limbic seizures as well as that the perirhinal cortex plays a preferential role in in vitro ictogenesis. PMID- 14751282 TI - Early increase of apoptosis-linked gene-2 interacting protein X in areas of kainate-induced neurodegeneration. AB - Apoptosis-linked gene-2 interacting protein X (Alix) is thought to be involved in both cell death and vesicular trafficking. We examined Alix expression 2 h, 6 h and 24 h after triggering seizure-dependent neuronal death by i.p. kainic acid injection. In the hippocampus, intense, transient immunolabelling was observed in the strata lucidum, oriens and radiatum, areas of high synaptic activity. The similarity of this distribution to those of synaptophysin and endophilin suggests a presynaptic localisation. Alix labelling was increased in neuronal cell bodies in kainate-sensitive regions before or concomitant with the first signs of oedema and/or neuronal eosinophilia. The increase persisted 24 h after kainate-injection in CA3 and the piriform cortex which are areas with massive swelling and numerous pyknotic neurons. This suggests that Alix may play an early role in the mechanisms leading to cell death. Taken together, our results suggest that Alix may be a molecular link between synaptic functioning and neuronal death. PMID- 14751283 TI - Neuroprotection by transforming growth factor-beta1 involves activation of nuclear factor-kappaB through phosphatidylinositol-3-OH kinase/Akt and mitogen activated protein kinase-extracellular-signal regulated kinase1,2 signaling pathways. AB - Prevention of neuronal apoptosis has been introduced as a new therapeutic strategy for neurodegenerative disorders. We have previously reported anti apoptotic effects of transforming growth factor-beta1 (TGF-beta1), a multifunctional cytokine, in models of cerebral ischemia and in cultured neurons and recently focused on the mechanisms underlying the anti-apoptotic effect of TGF-beta1. The anti-apoptotic transcriptional factor nuclear factor kappa B (NF kappaB) shows high impact in the cell survival function of multiple cytokines and growth factors. The present study explored whether NF-kappabeta is a target of TGF-beta1 and which signaling pathways involved in the activation of NF-kappabeta are triggered by TGF-beta1. We demonstrated that TGF-beta1 increased the transcriptional activity of NF-kappabeta in cultured hippocampal neurons in a time- and concentration-dependent manner. Furthermore, TGF-beta1 induced translocation of p65/NF-kappabeta to the nucleus and enhanced NF-kappabeta transcriptional activity in the presence of apoptotic stimuli. TGF-beta1-mediated NF-kappabeta activation was blocked by wortmannin and U0126, indicating the involvement of both phosphatidylinositol-3-OH kinase (PI3k)/Akt and mitogen activated protein kinase (MAPK)/extracellular-signal regulated kinase (Erk)1,2 pathways in the action of TGF-beta1. TGF-beta1 produced a concomitant increase in the phosphorylations of Ikappabeta kinase (IKKalpha/beta) and Ikappabetaalpha with a subsequent degradation of Ikappabetaalpha. Interestingly, the increased phosphorylation of IKKalpha/beta and Ikappabetaalpha was abrogated by wortmannin, but not by U0126, suggesting that PI3k/Akt and MAPK/Erk1,2 pathways triggered by TGF-beta1 regulated the activation of NF-kappabeta through different mechanisms. Of note, wortmannin and U0126, as well as kappabeta-decoy DNA, abolished the anti apoptotic effect of TGF-beta1, corroborating the notion that both PI3k/Akt and MAPK/Erk1,2 pathways, and NF-kappabeta activity are necessary for the anti apoptotic activity of TGF-beta1. PMID- 14751284 TI - Accumulation of Ym1/2 protein in the mouse olfactory epithelium during regeneration and aging. AB - A unique feature of the olfactory system is its efficiency to produce new neurons in the adult. Thus, destruction of the olfactory receptor neurons (ORNs) using chemical (intranasal perfusion with ZnSO4) or surgical (axotomy or bulbectomy) methods, leads to an enhanced rate of proliferation of their progenitors and to complete ORNs regeneration. The aim of our study was to identify new factors implied in this regenerative process. Using an electrophoretic method, we observed the accumulation of a 42 kDa protein after axotomy in the olfactory mucosa, but not in the olfactory bulb. Its expression started after a few days following injury and increased massively during the phase of ORN regeneration. The purification and the sequence characterization revealed that this protein was Ym1/2, recently identified in activated macrophages present in various tissues during inflammation. Western blotting analysis of Ym1/2 confirmed the accumulation of this protein in the regenerating olfactory mucosa consecutively to olfactory axotomy or bulbectomy but also after ZnSO4 irrigation of the nasal cavity. In the olfactory mucosa of control mice, Ym1/2 was hardly detectable in young animals and became more and more abundant with increasing age. In injured and aged mice, Ym1/2 mainly accumulates in the cytoplasm of supporting cells as well as in other cells located throughout the olfactory epithelium. Our results suggest that Ym1/2 is involved in olfactory epithelium remodeling following several kinds of lesions of the adult olfactory mucosa and support the view of a critical role of inflammatory cues in neurodegeneration and aging. PMID- 14751285 TI - Cellular localization of Shab and Shaw potassium channels in the lobster stomatogastric ganglion. AB - The motor pattern generated by the 14 neurons composing the pyloric circuit in the stomatogastric ganglion (STG) of the spiny lobster, Panulirus interruptus, is organized not only by the synaptic connections between neurons, but also by the characteristic intrinsic electrophysiological properties of the individual cells. These cellular properties result from the unique complement of ion channels that each cell expresses, and the distribution of those channels in the cell membranes. We have mapped the STG expression of shab and shaw, two genes in the Shaker superfamily of potassium channel genes that encode voltage-dependent, non inactivating channels. Using antibodies developed against peptide sequences from the two channel proteins, we explored the localization and cell-specific expression of the channels. Anti-Shab and anti-Shaw antibodies both stain all the pyloric neurons in the somata, as well as their primary neurites and branch points of large neurites, but to varying degrees between cell types. Staining was weak and irregular (Shaw) or absent (Shab) in the fine neuropil of pyloric neurons, where most synaptic interactions occur. There is a high degree of variability in the staining intensity among neurons of a single cell class. This supports Golowasch et al.'s [J Neurosci 19 (1999) RC33; Neural Comput 11 (1999) 1079] hypothesis that individual cells can have similar firing properties with varying compositions of ionic currents. Both antibodies stain the axons of the peripheral nerves as they enter foregut muscles. We conclude that both Shab and Shaw channels are appropriately localized to contribute to the noninactivating potassium current in the stomatogastric nervous system. PMID- 14751286 TI - TFII-I, a candidate gene for Williams syndrome cognitive profile: parallels between regional expression in mouse brain and human phenotype. AB - The gene for TFII-I, a widely expressed transcription factor, has been localized to an interval of human chromosome 7q11.23 that is commonly deleted in Williams syndrome (WS). The clinical phenotype of WS includes elfin facies, infantile hypercalcemia, supravalvular aortic stenosis, hyperacusis and mental retardation. The WS cognitive profile (WSCP) is notable for the differential impairment of visual-spatial abilities with relative sparing of verbal-linguistic function. Fine mapping of individuals with WS has revealed a close association between deletion of TFII-I and the WSCP. To determine the plausibility of the hypothesis that hemizygous deletion of TFII-I contributes to the WSCP, we have examined the anatomic distribution of TFII-I RNA and protein isoforms in brains from adult and embryonic mice. Our studies show that early in development, TFII-I expression is widespread and nearly uniform throughout the brain. In adult brain, TFII-I protein is present exclusively in neurons. Highest levels of expression are observed in cerebellar Purkinje cells and in hippocampal interneurons. TFII-I immunoreactivity is distinct from that of the related protein, TFII-IRD1, which is also localized to the region of human chromosome 7 deleted in WS. The expression pattern of TFII-I in mouse brain parallels regions in human brain which have been shown to be anatomically and functionally altered in humans with WS. These observations are consistent with the hypothesis that deletion of the gene for TFII-I contributes to the cognitive impairments observed in WS. PMID- 14751287 TI - Developmental expression of methyl-CpG binding protein 2 is dynamically regulated in the rodent brain. AB - The gene encoding methyl-CpG binding protein 2 (MeCP2) is mutated in the large majority of girls that have Rett Syndrome (RTT), an X-linked neurodevelopmental disorder. To better understand the developmental role of MeCP2, we studied the ontogeny of MeCP2 expression in rat brain using MeCP2 immunostaining and Western blots. MeCP2 positive neurons were present throughout the brain at all ages examined, although expression varied by region and age. At early postnatal ages, regions having neurons that were generated early and more mature had the strongest MeCP2 expression. Late developing structures including cortex, hippocampus and cerebellum exhibited the most significant changes in MeCP2 expression. Of these regions, the cerebellum showed the most striking cell specific changes in MeCP2 expression. For example, the early-generated Purkinje cells became MeCP2 positive by P6, while the late-generated granule cells did not express MeCP2 until the fourth postnatal week. The timing of MeCP2 expression in the granule cell layer is coincident with the onset of granule cell synapse formation. Although more subtle, the degree of MeCP2 expression in cortex and hippocampus was most closely correlated with synaptogenesis in both regions. Our finding that MeCP2 expression is correlated with synaptogenesis is consistent with the hypothesis that Rett Syndrome is caused by defects in the formation or maintenance of synapses. PMID- 14751288 TI - Expression of axon guidance molecules and their related genes during development and sexual differentiation of the olfactory bulb in rats. AB - Axon guidance molecules and related proteins such as semaphorin 3A, neuropilin-1, plexin-1, netrin-1, growth-associated protein, olfactory marker protein, cypin and collapsin response mediator proteins guide the development of neural circuits in the olfactory bulb. In this study, transcriptions of these genes were examined in the olfactory bulb of female, male and neonatal testosterone propionate treated female rats at the ages of 2, 5, 10, 15, 20, 25, 30 and 45 days. The semaphorin 3A, neuropilin-1, growth-associated protein and collapsin response mediator protein 1-5 genes were expressed significantly higher during the early development stages than in adulthood while the opposite is true for the olfactory marker protein. The expression profile of cypin and netrin-1 was relatively constant through development. A late effect of the neonatal testosterone propionate treatment on netrin-1, growth-associated protein, olfactory marker protein, collapsin response mediator proteins 1, 3, 4 and cypin gene expression was observed. The expression profiles of collapsin response mediator proteins and their related genes in the developing olfactory bulb confirmed most studies on the relationship between collapsin response mediator proteins and development in the brain. Sex differences of semaphorin 3A, neuropilin-1 as well as collapsin response mediator protein 3 at the early development stage and the late effect of neonatal testosterone propionate treatment on the expressions of netrin-1, growth associated marker protein, cypin and collapsin response mediator proteins 1, 3 and 5 genes may indicate a possible role of these molecules on sexual differentiation of the olfactory bulb. PMID- 14751289 TI - Striatal phosphodiesterase mRNA and protein levels are reduced in Huntington's disease transgenic mice prior to the onset of motor symptoms. AB - Inheritance of a single copy of the gene encoding huntingtin (HD) with an expanded polyglutamine-encoding CAG repeat leads to neuronal dysfunction, neurodegeneration and the development of the symptoms of Huntington's disease (HD). We have found that the steady-state mRNA levels of two members of the phosphodiesterase (PDE) multi-gene family decrease over time in the striatum of R6 transgenic HD mice relative to age-matched wild-type littermates. Phosphodiesterase 10A (PDE10A) mRNA and protein levels decline in the striatum of R6/1 and R6/2 HD mice prior to motor symptom development. The rate of reduction in PDE10A protein correlates with the rate of decline of the message and the decrease in PDE10A mRNA and protein is more rapid in R6/2 compared with R6/1 mice. Both PDE10A protein and mRNA, therefore, decline to minimum levels prior to the onset of overt physical symptoms in both strains of transgenic mice. Moreover, protein levels of PDE10A are decreased in the caudate-putamen of grade 3 HD patients compared with age-matched neuropathologically normal controls. Striatal PDE1B mRNA levels also decline in R6/1 and R6/2 HD mice; however, the decrease in striatal PDE10A levels (>60%) was greater than that observed for PDE1B and immediately preceded the onset of motor symptoms. In contrast, PDE4A mRNA levels are relatively low in the striatum and do not differ between age matched wild-type and transgenic HD mice. This suggests that the regulation of PDE10A and PDE1B, but not PDE4A, mRNA levels is dependent on the relative expression of or number of CAG repeats within the human HD transgene. The loss of phosphodiesterase activity may lead to dysregulation of cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) levels in the striatum, a region of the brain that contributes to the control of movement and cognition. PMID- 14751290 TI - Localization of VGLUT3, the vesicular glutamate transporter type 3, in the rat brain. AB - We have recently identified a third subtype of glutamate vesicular transporter (VGLUT) named VGLUT3. In the present study, we provide a detailed account of the regional and cellular distributions of VGLUT3 in the rat brain, using specific nucleotide probes and antisera. The distribution of VGLUT3 protein was compared with that of the other vesicular transporters (VGLUT1 and VGLUT2). All the areas expressing VGLUT3 also contain high levels of VGLUT1 and -2 proteins, but, at a finer level of analysis, the distribution of the three subtypes differs. Unlike VGLUT1 and -2, VGLUT3 expression is limited to discrete cell populations. Neurons containing VGLUT3 transcript are essentially observed in the caudate-putamen, the olfactory tubercle, the nucleus accumbens, the hippocampus, the interpeduncular nucleus and the dorsal and medial raphe nuclei. More scattered populations of VGLUT3 expressing neurons are found in the cerebral cortex. The distribution of the VGLUT3 protein, as determined with specific antisera, overlaps with that of the transcript in the caudate-putamen, olfactory tubercles, hippocampus, cortex, interpeduncular nucleus, and raphe nuclei, suggesting that VGLUT3 is essentially present in local projection neurons in these regions. Microscopic examination reveals staining of terminals and perikarya. Furthermore, co-localization studies indicate that VGLUT3 is present in GABAergic interneurons in the hippocampus, as well as in the interpeduncular nucleus. However, other regions, such as the substantia nigra (pars compacta), the ventral tegmental area, and the parabigeminal nucleus, receive a dense VGLUT3 terminal labeling although they do not contain VGLUT3 expressing neurons. In these regions, VGLUT3 immunoreactivity may be present in terminals of long projecting neurons. This subclass of glutamatergic afferents differs from other "classical" excitatory terminals that express VGLUT1 or VGLUT2. The distribution of VGLUT3 in the rat brain suggests an unsuspected function of vesicular glutamate transport in subsets of interneurons and in neuromodulatory neurons. PMID- 14751291 TI - Effects of chronic cocaine exposure on corticotropin-releasing hormone binding protein in the central nucleus of the amygdala and bed nucleus of the stria terminalis. AB - The neuropeptide, corticotropin-releasing hormone (CRH), has been shown to play a role in behavioral and neurobiological effects of drugs of abuse. An important modulator of CRH, the CRH binding protein (CRH-BP), has not, on the other hand, been assessed for its role in drug-associated effects. The primary objective of the present experiment was to assess whether prior, chronic exposure to cocaine modulates expression of CRH-BP, and to compare expression of the BP with that of the peptide itself. We assessed CRH-BP and CRH mRNA expression in two brain regions where CRH is known to affect responses to drugs of abuse; namely, the central nucleus of the amygdala (CeA) and bed nucleus of the stria terminalis (BNST). Male Long-Evans rats were given 14 daily injections of cocaine (30 mg/kg, i.p.) or saline. One, 3, 10, 28, or 42 days post-treatment, animals were killed and adjacent brain sections through the CeA and BNST were processed for CRH-BP and CRH by in situ hybridization. In the CeA, cocaine pre-exposure increased both CRH and CRH-BP mRNA expression 1 day post-treatment. In the dorsal BNST, cocaine pre-exposure elevated levels of CRH-BP, but not CRH, mRNA 3 days post-treatment. Taken together, the results suggest that withdrawal-induced changes in the expression of the CRH-BP, and CRH itself, are relatively short-lived and that a dysregulation in basal expression of either gene is not likely responsible for long-lasting behavioral effects noted with cocaine and other drugs of abuse. PMID- 14751292 TI - Modulation of AMPA receptor kinetics differentially influences synaptic plasticity in the hippocampus. AB - Prior studies showed that positive alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionic acid (AMPA) receptor modulators facilitate long-term potentiation (LTP) and improve the formation of several types of memory in animals and humans. However, these modulators are highly diverse in their effects on receptor kinetics and synaptic transmission and thus may differ also in their efficacy to promote changes in synaptic strength. The present study examined three of these modulators for their effects on synaptic plasticity in field CA1 of hippocampal slices, two of them being the benzamide drugs 1-(quinoxalin-6 ylcarbonyl)piperidine (CX516) and 1-(1,4-benzodioxan-6-ylcarbonyl)piperidine (CX546) which prominently enhance synaptic transmission yet differ in their relative impact on amplitude versus duration of the synaptic response. The third drug was cyclothiazide which potently blocks AMPA receptor desensitization. Effects on plasticity were assessed by measuring (i) the likelihood of obtaining stable potentiation when using theta-burst stimulation with three instead of four pulses per burst, (ii) the maximum amount of potentiation under optimal stimulation conditions, and (iii) the effect on long-term depression (LTD). Both benzamides facilitated the formation of stable potentiation induced with three pulse burst stimulation which is normally ineffective. CX546 in addition increased maximally inducible potentiation after four-pulse burst stimulation from about 50% to 100%. Burst response analysis revealed that CX546 greatly prolonged the duration of depolarization by slowing the decay of the response which thus presumably leads to a more continuous N-methyl-D-aspartate (NMDA) receptor activation. Cyclothiazide was ineffective in increasing maximal potentiation in either field or whole-cell recordings. CX546, but not CX516, also enhanced nearly two-fold the NMDA receptor-dependent long-term depression induced by heterosynaptic 2 Hz stimulation. Tests with recombinant NMDA receptors (NR1/NR2A) showed that CX516 and CX546 have no direct effects on currents mediated by these receptors. These results suggest that (1) modulation of AMPA receptors which increases either response amplitude or duration can facilitate LTP formation, (2) modulators that effectively slow response deactivation augment the maximum magnitude of LTP and LTD, and (3) receptor desensitization may have a minor impact on synaptic plasticity in the hippocampus. Taken together, our data indicate that AMPA receptor modulators differ substantially in their ability to enhance synaptic potentiation or depression, depending on their particular influence on receptor kinetics, and hence that they may also be differentially effective in influencing higher-order processes such as memory encoding. PMID- 14751293 TI - Stimulation of adenosine A2A receptors elicits zif/268 and NMDA epsilon2 subunit mRNA expression in cortex and striatum of the "weaver" mutant mouse, a genetic model of nigrostriatal dopamine deficiency. AB - Interaction between basal ganglia and cerebral cortex is critical for normal goal directed behavior. In the present study we have used the immediate early gene zif/268, as functional marker to investigate how the stimulation of adenosine A2A receptors, i.e. of the "indirect" striatal output pathway, affects striatal and cortical function in "weaver" mouse, a genetic model of dopamine deficiency. Furthermore, we have examined the effect of A2A receptor stimulation on glutamate receptor expression in the "weaver" brain. A single injection of CGS21680 (A2A receptor agonist), induced strong expression of zif/268 mRNA, detected by in situ hybridization, not only in striatum but also in the motor cortex of the "weaver" mutant. This cortical response seems to be elicited through the basal-ganglia thalamo-cortical circuit, rather than through a direct cortical effect, since A2A receptors are not detectable in cortex according to our autoradiographic study. Co-administration of CGS21680 and quinpirole (D2 receptor agonist) attenuated the expression of zif/268 mRNA in dorsal striatum but not in motor cortex, indicating that the cortical response is dopamine-D2-receptor-independent. However, this co administration induced an increase in zif/268 mRNA expression in somatosensory cortex, which could rely on disinhibition of the thalamo-cortical pathway. The motor cortical response could be of clinical interest, as it would further stimulate the "indirect" striatal pathway in a feed forward circuit, thus worsening the parkinsonian symptoms. Furthermore, the up-regulation of epsilon2 subunit mRNA of the NMDA receptor, induced by CGS21680 administration, seen in striatum and cortex of the "weaver" mouse, would lead to overactivity of these receptors worsening dyskinesias. These results suggest adenosine to play a significant role in regulating striatal and cortical neurochemistry in a dopamine depleted mouse. Blockade of these receptors by specific A2A antagonists could ameliorate parkinsonian symptoms. PMID- 14751294 TI - Modulation of activator protein 1/DNA binding activity by acoustic overstimulation in the guinea-pig cochlea. AB - Changes in gene expression are part of the homeostatic machinery with which cells respond to external stimuli or assaults. The activity of the early response transcriptional factor activator protein-1 (AP-1) can be modulated by a variety of environmental stimuli including those that alter the cellular oxidation/reduction status. This study investigates the activation of AP-1/DNA binding in the guinea-pig cochlea in response to acoustic overstimulation which produces reactive oxygen species. Electrophoretic mobility shift assays revealed that binding of AP-1 to its radiolabeled oligonucleotide probe markedly changed in nuclear extracts of inner ear tissues following intense noise exposure (4 kHz octave band, 115 dB, 5 h). AP-1/DNA binding increased in the organ of Corti and the lateral wall tissues immediately after the exposure, returning to near baseline levels 5 h later. At 15 h after noise, a second peak of binding activity occurred in the organ of Corti whereas stria vascularis showed a lesser but more sustained activity. Binding in nuclear extracts from the spiral ganglion did not change. Incubation of nuclear extracts with antibodies against Fos/Jun family proteins prior to a supershift assay showed Fra-2 as a major component of the AP 1 complex immediately after the noise exposure. In the organ of Corti, Fra-2 immunoreactivity was localized to the middle turn, i.e. the region which is most affected by the 4-kHz octave band exposure. The results suggest the modulation of gene expression via the activation of AP-1 as a consequence of noise trauma but also demonstrate differential responses in cochlear tissues. PMID- 14751295 TI - Evidence of neuronal excitatory amino acid carrier 1 expression in rat dorsal root ganglion neurons and their central terminals. AB - The expression and distribution of the neuronal glutamate transporter, excitatory amino acid carrier-1 (EAAC1), are demonstrated in the dorsal root ganglion neurons and their central terminals. Reverse transcriptase-polymerase chain reaction shows expression of EAAC1 mRNA in the dorsal root ganglion. Immunoblotting analysis further confirms existence of EAAC1 protein in this region. Immunocytochemistry reveals that approximately 46.6% of the dorsal root ganglion neurons are EAAC1-positive. Most EAAC1-positive neurons are small and around 250-750 microm2 in surface area, and some co-label with calcitonin gene related peptide (CGRP) or isolectin IB4. In the spinal cord, EAAC-1 immunoreactive small dot- or patch-like structures are mainly localized in the superficial dorsal horn, and some are positive for CGRP or labeled by isolectin IB4. Unilateral dorsal rhizotomy experiments further show that EAAC1 immunoreactivity is less intense in superficial dorsal horn on the side ipsilateral to the dorsal rhizotomy than on the contralateral side. The results indicate the presence of EAAC1 in the dorsal root ganglion neurons and their central terminals. Our findings suggest that EAAC1 might play an important role in transmission and modulation of nociceptive information via the regulation of pre-synaptically released glutamate. PMID- 14751296 TI - N-acetylaspartate levels of left frontal cortex are associated with verbal intelligence in women but not in men: a proton magnetic resonance spectroscopy study. AB - The left frontal cortex plays an important role in executive function and complex language processing inclusive of spoken language. The purpose of this work was to assess metabolite levels in the left and right prefrontal cortex and left anterior cingulum by proton magnetic resonance spectroscopy and relate results to verbal intelligence (Wechsler Adult Intelligence Scale revised) in a sample of college-educated healthy volunteers (dorsolateral prefrontal cortex [DLPFC]: n=52, 23 females, and left anterior cingulum: n=62, 22 females; age range: 20-75 years). In women only, N-acetylaspartate in the DLPFC and in the left anterior cingulate cortex was positively correlated with vocabulary scores. Our data support the hypothesis of existing gender differences regarding the involvement of the left frontal cortex in verbal processing as reflected in different correlations of specific metabolites with verbal scores. PMID- 14751298 TI - Horseradish peroxidase: a modern view of a classic enzyme. AB - Horseradish peroxidase is an important heme-containing enzyme that has been studied for more than a century. In recent years new information has become available on the three-dimensional structure of the enzyme and its catalytic intermediates, mechanisms of catalysis and the function of specific amino acid residues. Site-directed mutagenesis and directed evolution techniques are now used routinely to investigate the structure and function of horseradish peroxidase and offer the opportunity to develop engineered enzymes for practical applications in natural product and fine chemicals synthesis, medical diagnostics and bioremediation. A combination of horseradish peroxidase and indole-3-acetic acid or its derivatives is currently being evaluated as an agent for use in targeted cancer therapies. Physiological roles traditionally associated with the enzyme that include indole-3-acetic acid metabolism, cross-linking of biological polymers and lignification are becoming better understood at the molecular level, but the involvement of specific horseradish peroxidase isoenzymes in these processes is not yet clearly defined. Progress in this area should result from the identification of the entire peroxidase gene family of Arabidopsis thaliana, which has now been completed. PMID- 14751299 TI - On the origins of triterpenoid skeletal diversity. AB - The triterpenoids are a large group of natural products derived from C(30) precursors. Nearly 200 different triterpene skeletons are known from natural sources or enzymatic reactions that are structurally consistent with being cyclization products of squalene, oxidosqualene, or bis-oxidosqualene. This review categorizes each of these structures and provides mechanisms for their formation. PMID- 14751300 TI - Cyanogenic glucosides and plant-insect interactions. AB - Cyanogenic glucosides are phytoanticipins known to be present in more than 2500 plant species. They are considered to have an important role in plant defense against herbivores due to bitter taste and release of toxic hydrogen cyanide upon tissue disruption. Some specialized herbivores, especially insects, preferentially feed on cyanogenic plants. Such herbivores have acquired the ability to metabolize cyanogenic glucosides or to sequester them for use in their predator defense. A few species of Arthropoda (within Diplopoda, Chilopoda, Insecta) are able to de novo synthesize cyanogenic glucosides and, in addition, some of these species are able to sequester cyanogenic glucosides from their host plant (Zygaenidae). Evolutionary aspects of these unique plant-insect interactions with focus on the enzyme systems involved in synthesis and degradation of cyanogenic glucosides are discussed. PMID- 14751301 TI - Purification and identification of a Ca(2+)-pectate binding peroxidase from Arabidopsis leaves. AB - A protein fraction was obtained from Arabidopsis (Arabidopsis thaliana, L.) leaf extract by affinity chromatography through a Ca(2+)-pectate/polyacrylamide gel. Further purification by preparative isoelectric focusing and SDS PAGE allowed the separation of a peroxidase that was identified as being peroxidase AtPrx34 (AtprxCb, accession number X71794) by N-terminal amino acid microsequencing. AtPrx34 belongs to a group of five Arabidopsis sequences encoding putative pectin binding peroxidases. An expression study showed that it is expressed in root, stem, flower and leaf. It was produced by Escherichia coli and tested for its ability to bind to Ca(2+)-pectate. The identity of the amino acids involved in the interaction between the peroxidase and the Ca(2+)-pectate structure is discussed. PMID- 14751303 TI - Carbon and hydrogen isotopic fractionation during lipid biosynthesis in a higher plant (Cryptomeria japonica). AB - Compound-specific carbon and hydrogen isotopic compositions of lipid biomolecules (n-alkanes, n-alkanoic acids, n-alkanols, sesquiterpenes, diterpenes, phytol, diterpenols and beta-sitosterol), extracted from Cryptomeria japonica leaves, were determined in order to understand isotopic fractionations occurring during lipid biosynthesis in this species. All lipid biomolecules were depleted in both 13C and D relative to bulk tissue and ambient water, respectively. n-Alkyl lipids associated with the acetogenic pathway were depleted in 13C relative to bulk tissue by 2.4-9.9 per thousand and depleted in D relative to ambient water by 91 152 per thousand. C(15)- and C(30)-isoprenoid lipids (sesquiterpenes, squalene and beta-sitosterol) associated with the mevalonic-acid pathway are depleted in 13C relative to bulk tissue by 1.7-3.1 per thousand and depleted in D relative to ambient water by 212-238 per thousand. C(20)-isoprenoid lipids (phytol and diterpenoids) associated with the non-mevalonic-acid pathway were depleted in 13C relative to bulk tissue by 4.6-5.9 per thousand and depleted in D relative to ambient water by 238-303 per thousand. Phytol was significantly depleted in D by amounts up to 65 per thousand relative to other C(20) isoprenoid lipids. The acetogenic, mevalonic-acid and non-mevalonic-acid pathways were clearly discriminated using a cross-plot between the carbon and hydrogen isotopic fractionations. PMID- 14751302 TI - Signatures of cinnamyl alcohol dehydrogenase deficiency in poplar lignins. AB - A series of transgenic poplars down-regulated for cinnamyl alcohol dehydrogenase (CAD) was analyzed by thioacidolysis. Among the lignin-derived monomers, the indene compounds that were recently shown to originate from sinapaldehyde incorporated into lignins through 8-O-4-cross-coupling, were found to increase as a function of CAD deficiency level. While these syringyl markers were recovered in substantial amounts in the most severely depressed lines, the markers for coniferaldehyde incorporation were recovered in only low amounts. In conjunction with these additional sinapaldehyde units and relative to the control samples, lignins in CAD-deficient poplar lines had less conventional syringyl-units and beta-O-4-bonds and more free phenolic groups. We found that almost half of the polymers in the most deficient lines could be solubilized in alkali and at room temperature. This unusual behavior suggests that lignins in CAD-deficient poplars occur as small, alkali-leachable lignin domains. That mainly sinapaldehyde incorporates into the lignins of CAD-deficient poplars suggests that the recently identified sinapyl alcohol dehydrogenase (SAD), which is structurally distinct from the CAD enzyme targeted herein, does not play any substantial role in constitutive lignification in poplar. PMID- 14751304 TI - The anti-staphylococcal activity of Angelica dahurica (Bai Zhi). AB - Bioassay-guided fractionation of a hexane extract prepared from the roots of the Chinese drug Angelica dahurica (Bai Zhi) led to the isolation of the polyacetylenic natural product falcarindiol (1). The absolute stereochemistry of this compound was confirmed by careful 1H NMR analysis of its (R)- and (S)-Mosher ester derivatives as the 3(R), 8(S) isomer. Activity was tracked using a Mycobacterium fortuitum screening assay and the purified product was evaluated against multidrug-resistant and methicillin-resistant strains of Staphylococcus aureus (MRSA). The minimum inhibitory concentrations (MIC) of this metabolite ranged from 8 to 32 microg/ml highlighting the potential of the acetylene natural product class as antibiotic-lead compounds. These MIC values compare favourably with some of the newest agents in development for the treatment of MRSA infection and indicate that further evaluation of the antibiotic activity of acetylenes is warranted. PMID- 14751305 TI - Generation of ginsenosides Rg3 and Rh2 from North American ginseng. AB - Rg3 and Rh2 ginsenosides are primarily found in Korean red ginseng root (Panax ginseng C.A. Meyer) and valued for their bioactive properties. We quantified both Rh2 and Rg3 ginseng leaf and Rg3 from root extracts derived from North American ginseng (Panax quinquefolius). Quantification was obtained by application of HPLC with ion fragments detected using ESI-MS. Ginseng leaf contained 11.3+/-0.5 mg/g Rh2 and 7.5+/-0.9 mg/g Rg3 in concentrated extracts compared to 10.6+/-0.4 mg/g Rg3 in ginseng root. No detectable Rh2 was found in root extracts by HPLC, although it was detectable by ESI-MS analysis. Ginsenosides Rg3 and Rh2 were detected following hot water reflux extraction, but not from tissues extracted with 80% aqueous ethanol at room temperature. Therefore ginsenosides Rg3 and Rh2 are not naturally present in North American ginseng, but are products of a thermal process. Using ESI-MS analysis, it was found that formation of Rg3 and Rh2, among other compounds, were a function of heating time and were breakdown products of the more abundant ginsenosides Rb1 and Rc. Our findings that heat processed North American ginseng leaf is an excellent source of Rh2 ginsenoside is an important discovery considering that ginseng leaf material is obtainable throughout the entire plant cycle for recovery of valuable ginsenosides for pharmaceutical use. PMID- 14751306 TI - Potential cancer chemopreventive constituents of the leaves of Macaranga triloba. AB - Activity-guided fractionation of the leaves of Macaranga triloba, using an in vitro bioassay based on the inhibition of cyclooxygenase-2, resulted in the isolation of a rotenoid, 4,5-dihydro-5'alpha-hydroxy-4'alpha-methoxy-6a,12a dehydro-alpha-toxicarol (1), as well as 12 known compounds, (+)-clovan 2beta,9alpha-diol, ferulic acid, 3,7,3',4'-tetramethylquercetin, 3,7,3' trimethylquercetin, 3,7-dimethylquercetin, abscisic acid, 1beta,6alpha-dihydroxy 4(15)-eudesmene, 3beta-hydroxy-24-ethylcholest-5-en-7-one, loliolide, scopoletin, taraxerol, and 3-epi-taraxerol. The structure of compound 1 was determined using spectroscopic methods. All isolates were evaluated for their potential to inhibit cyclooxygenases-1 and -2 by measuring PGE(2) production, and to induce quinone reductase in cultured Hepa 1c1c7 mouse hepatoma cells. PMID- 14751308 TI - Oligomeric hydrolyzable tannins from Monochaetum multiflorum. AB - Four hydrolyzable tannins, nobotanins Q, R, S, and T, were isolated from the aqueous acetone extract of the dried leaves of Monochaetum multiflorum (Melastomataceae), a plant indigenous to Colombia. Their dimeric and tetrameric structures were elucidated by spectral and chemical methods. Eight known hydrolyzable tannin monomers and eight ellagitannin oligomers characteristic of melastomataceous plants were also characterized as tannin constituents of the plant. PMID- 14751307 TI - Cinnamoyl glucosides of catechin and dimeric procyanidins from young leaves of Inga umbellifera (Fabaceae). AB - The rapidly growing, nearly achlorophyllous, young leaves of Inga umbellifera express high concentrations of mono and dimeric 3-O-gluco-cinnamoyl catechin/epicatechin, rare forms of substituted flavan-3-ols. Here we present structures for five novel compounds in this class: three monomers [catechin-3-O beta-D-gluco(2-cinnamoyl)pyranoside, catechin-3-O-beta-D-gluco(6-cinnamoyl) pyranoside, catechin-3-O-beta-D-gluco(2,6-biscinnamoyl)pyranoside] and two dimeric procyanidins [catechin-3-O-beta-D-glucopyrano-(4alpha-->8)-catechin-3-O beta-D-gluco(2-cinnamoyl)pyranoside and catechin-3-O-beta-D-glucopyrano-(4alpha- >8)-epicatechin-3-O-beta-D-gluco(6-cinnamoyl)pyranoside]. The young leaves of Inga umbellifera express high concentrations of 3-O-(cinnamoyl)glucosides of catechin and epicatechin. PMID- 14751309 TI - Slow release of two antibiotics of veterinary interest from PVA hydrogels. AB - Two antibiotics, tylosin tartrate and oxytetracycline hydrochloride, were entrapped in poly(vinyl alcohol) (PVA) hydrogels (MW 31,000-50,000) by a cryogen procedure obtaining a controlled release system suitable for veterinary application. It was found that at a low drug matrix loading (10 mg/ml), the in vitro release rate of both antibiotics could be reduced by a previous freeze drying of the gel, while no reduction in drug rate took place in heavily loaded matrices (300 mg/ml). When PVA hydrogels containing tylosin were administered to rats per os the drug could not be detected in the blood, but it was found in organs,: liver, kidneys, and muscles, for up to 120 h. On the other hand, when the same amount of drug was administered orally as powder, no appreciable organ accumulation was detected, while the drug was found in faeces and urine. These data show that PVA hydrogels can be a suitable slow release system for tylosin administration. Oxytetracycline could also be quantitatively entrapped and released from PVA hydrogels, but once administered per os to rats, it was not detected in blood or organs. PMID- 14751310 TI - Synthesis and anticonvulsant properties of tetrahydroisoquinoline derivatives. AB - As a follow up of our previous structure-activity relationship and molecular modeling studies, we synthesized a novel series of 1-aryl-6,7-dimethoxy-1,2,3,4 tetrahydroisoquinoline derivatives as potential non-competitive AMPA receptor antagonists. When tested for their ability to prevent sound-induced seizures in DBA/2 mice, some of these novel compounds showed high anticonvulsant potency. PMID- 14751311 TI - Biodegradation of DNA and nucleotides to nucleosides and free bases. AB - Thirty-two different microorganisms were examined in order to check their ability to degrade an exogenous DNA. Bacteria from species: Stenotrophomonas maltophilia, Brevundimonas diminuta, Bacillus subtilis, Mycobacterium butyricum and fungus Fusarium moniliforme were capable to degrade DNA to nucleic bases or their derivatives. Degradation of DNA by S. maltophilia resulted in formation of free bases, such as hypoxanthine, thymine, uracil and xanthine. The optimum concentration of DNA seemed to be 3 mg ml(-1). The mode of degradation of DNA nucleotides depended on the type of nucleotide and its concentration, but nucleic bases or their derivatives were always formed at the end of the reaction process. PMID- 14751312 TI - Direct spectrophotometric determination of quercetin in the presence of ascorbic acid. AB - The current research provides a simplified sample preparation procedure for the accurate estimation of quercetin in pure and in the pharmaceutical dosage form. Direct spectrophotometric method for the determination of quercetin in the presence of ascorbic acid was established. The influences of medium, wavelength, pH, temperature and the ionic strengths on quercetin determination were investigated. The best conditions for calibration curve are: 50% ethanol, lambda = 370 nm, pH = 4.2, T = 34 degrees C and I = 7.5 x 10(-5) M. Beer's law is obeyed in the concentration range 1.0-12.0 microg ml(-1) for quercetin. The corresponding detection limit is 0.76 microg ml(-1). The proposed method was verified by analyzing Quercetin + C capsules, Twinlab. PMID- 14751313 TI - Synthesis and COX-2 inhibitory properties of N-phenyl- and N-benzyl-substituted amides of 2-(4-methylsulfonylphenyl)cyclopent-1-ene-1-carboxylic acid and of their pyrazole, thiophene and isoxazole analogs. AB - Some N-phenyl- (7a-10a) and N-benzyl-substituted (7b-10b) amido analogs of cyclooxygenase (COX-2) selective tricyclic non-steroidal anti-inflammatory drugs have been synthesized with the aim to obtain information on the structural requirements for the COX-inhibitory activity. Compounds 7-10 were tested in vitro for their inhibitory properties only towards COX-2 enzyme by measuring prostaglandin E2 (PGE2) production on activated J774.2 macrophages. Some of the new compounds (7a, 8a, 9a and 9b) showed a modest activity, with percentage inhibition values near 30% at a concentration of 10 microM. These data have been tentatively explained by a conformational study which indicates that at least the N-phenyl-substituted amides 7a-9a present steric hindrances which may prevent a good interaction with COX-2 active site. PMID- 14751314 TI - Synthesis of new 2,3-diaryl-1,3-thiazolidin-4-ones as anti-HIV agents. AB - Several 2,3-diaryl-1,3-thiazolidin-4-ones were synthesized and evaluated as anti HIV agents. The results of the in vitro tests showed that some of them were highly effective inhibitors of HIV-1 replication at 30-50 nM concentrations with minimal cytotoxicity, thereby acting as non-nucleoside HIV-1 reverse transcriptase inhibitors (NNRTIs). PMID- 14751315 TI - DNA-directed alkylating agents: synthesis, antitumor activity and DNA affinity of bis-N,N'-trisubstituted 1,2,4-triazolo-piperazines. AB - A series of 1,4-bis-(1,5-dialkyl-1H-1,2,4-triazol-ylmethyl)piperazines and N methyl-piperazine analogs were prepared spontaneously from the cycloaddition of various reactive cumulenes with the piperazino-1,4-(bis-ethanenitrile) and 1 cyanomethyl-4-methyl-piperazine, respectively. The new compounds were evaluated for their DNA affinity and antitumor activity. PMID- 14751316 TI - Extractive spectrophotometric methods for the determination of nifedipine in pharmaceutical formulations using bromocresol green, bromophenol blue, bromothymol blue and eriochrome black T. AB - Four simple, sensitive and accurate spectrophotometric methods have been developed for the determination of nifedipine in pharmaceutical formulations. These methods are based on the formation of ion-pair complexes of amino derivative of the nifedipine with bromocresol green (BCG), bromophenol blue (BPB), bromothymol blue (BTB) and eriochrome black T (EBT) in acidic medium. The coloured products are extracted with chloroform and measured spectrophotometrically at 415 nm (BCG, BPB and BTB) and 520 nm (EBT). Beer's law was obeyed in the concentration range of 5.0-32.5, 4.0-37.5, 6.5-33.0 and 4.5 22.5 microg ml(-1) with molar absorptivity of 6.41 x 10(3), 4.85 x 10(3), 5.26 x 10(3) and 7.69 x 10(3) l mol(-1) cm(-1) and relative standard deviation of 0.82%, 0.72%, 0.66% and 0.68% for BCG, BPB, BTB and EBT methods, respectively. These methods have been successfully applied for the assay of drug in pharmaceutical formulations. No interference was observed from common pharmaceutical adjuvants. Statistical comparison of the results with the reference method shows excellent agreement and indicates no significant difference in accuracy and precision. PMID- 14751317 TI - Analgesic and anti-inflammatory activities evaluation of (-)-O-acetyl, (-)-O methyl, (-)-O-dimethylethylamine cubebin and their preparation from (-)-cubebin. AB - The anti-inflammatory and antinociceptive effects of the acetylated (2), methylated (3) and aminated (4) derivatives of cubebin (1), obtained by its reaction with acetic anhydride, methyl iodide and dimethylethylamine chloride, respectively, were investigated, using different animal models. The compound (2) was the most effective anti-inflammatory one in the carrageenin-induced paw edema in rats and was the only one which showed dose-response correlation for this assay with r = 0.993 and Y = 64.58x + 0.22. Besides, compounds (2) and (4) were more effective than cubebin in inhibiting acetic acid-induced writhing in mice, producing dose-response correlation with doses of 10, 20 and 30 mg/kg, respectively. Regarding the hot plate and the cell migration tests in rats, none of the four tested compounds showed activity. Overall, the results showed that the acetylation and amination of cubebin were efficient in enhancing its analgesic activity, as well as its anti-inflammatory activity. PMID- 14751318 TI - Spectrophotometric determination of gliclazide in pharmaceuticals and biological fluids through ternary complex formation with eosin and palladium (II). AB - A simple and sensitive spectrophotometric method has been developed for the determination of gliclazide (GLZ) in pharmaceutical formulations and biological fluids. The proposed method is based upon the formation of a ternary complex between palladium (II), eosin and GLZ in the presence of methyl cellulose as a surfactant and acetate buffer of pH 4.5. The ternary complex showed an absorption maximum at 550 nm. The solution of ternary complex obeyed Beer's law over the concentration range of 0.5-4 microg ml(-1) with minimum detectability (S/N = 2) of 0.05 microg ml(-1) (1.545 x 10(-7) M). The different experimental parameters affecting the development and stability of the color were carefully studied and optimized. The proposed method was successfully applied to the analysis of commercial tablets. The results obtained were in good agreement with those obtained using the official or reference spectrophotometric method. The proposed method was further applied to spiked human urine and plasma, the percentage recoveries were 97.84 +/- 0.72 and 97.43 +/- 0.83, respectively, (n = 4). A proposal of the reaction pathway was presented. PMID- 14751319 TI - Physician and enrollee knowledge of Medicaid coverage for tobacco dependence treatments. AB - BACKGROUND: The 2000 Public Health Service Clinical Practice Guideline, Treating Tobacco Use and Dependence, recommends health insurance coverage for tobacco dependence treatments proven effective in helping smokers to quit. Two states with comprehensive coverage for tobacco-dependence treatments in their Medicaid programs were selected to document awareness of coverage for tobacco-dependence treatments among primary care physicians who treat Medicaid enrollees and Medicaid-enrolled smokers. METHODS: In 2000, surveys were conducted among Medicaid smokers (n =400) and physicians (n =160) to document knowledge of covered tobacco-dependence treatments under state Medicaid programs in two states with comprehensive coverage. RESULTS: Only 36% of Medicaid-enrolled smokers and 60% of Medicaid physicians knew that their state Medicaid program offered any coverage for tobacco-dependence treatments. Physicians were more than twice as likely to know that pharmacotherapies were covered compared to counseling. CONCLUSIONS: Greater effort is needed to make Medicaid smokers and physicians aware that effective pharmacotherapies and counseling services are available to assist in treating tobacco dependence. Additionally, future research should explore the methods that are most effective in informing patients and providers regarding covered benefits. PMID- 14751320 TI - Latinos report less use of pharmaceutical aids when trying to quit smoking. AB - BACKGROUND: Latino smokers are more likely than white non-Latino smokers to attempt cessation, but less likely to receive cessation advice from physicians or to use nicotine replacement therapy (NRT). Proposed underlying causes have included lighter smoking, lower financial status, and less healthcare access. This study assessed these factors as possible explanations for disparate rates of smoking-cessation support. METHODS: Data were analyzed from a random, population level telephone survey of Colorado adults that interviewed 10,945 white non Latino respondents and 1004 Latino respondents. For the current analysis, main outcome measures were receipt of physician advice to quit smoking, use of NRT, and use of bupropion or other anti-depressant for smoking cessation. RESULTS: Latino smokers reported higher prevalence of quit attempts (71.5% v 61.6%, p <0.01) but less physician advice to quit smoking (46.4% v 56.2%, p <0.05) and less use of NRT or an anti-depressant for cessation (10.6% v 24.8%, p <0.0001). Adjusted for potentially confounding factors, the odds ratio (OR) for less Latino use of cessation medications was substantial and significant (full model OR=0.31; 95% confidence interval, 0.17 to 0.57). The adjusted OR for physician cessation advice was not significant. CONCLUSIONS: Population-level differences in health status, smoking level, financial status, or healthcare access do not explain why Latino smokers less often use proven pharmaceutical aids to increase cessation. Further research is needed to understand these disparities, and greater effort is needed to deliver cessation support to Latino smokers seeking to quit. PMID- 14751321 TI - Inpatient smoking-cessation counseling and all-cause mortality among the elderly. AB - BACKGROUND: Although smoking cessation is essential to the management of acute myocardial infarction (AMI), prevalence and benefits of smoking-cessation counseling in the inpatient setting are not well described among older adults. The objective of this study was to evaluate associations between inpatient smoking-cessation counseling and 5-year all-cause mortality among older adults hospitalized with AMI. METHODS: The Cooperative Cardiovascular Project (January 1994-July 1995) included 788 Medicare beneficiaries aged >/=65 years who were current smokers, admitted to acute care facilities in North Carolina with confirmed AMI, and discharged alive. Information on smoking-cessation advice or counseling prior to discharge was abstracted from medical records. Associations of counseling with 5-year risk of death were assessed with multivariate Cox proportional hazards regression. RESULTS: Smoking-cessation counseling was provided to 40% of AMI patients before discharge. Women (p =0.06) and blacks (p =0.02) were less likely to receive counseling. Counseling was associated with a history of chronic obstructive pulmonary disease (p =0.01). Increasing age, discharge to a skilled nursing facility, and histories of hypertension, heart failure, or stroke were associated with no counseling (p <0.05, all cause). Age adjusted mortality rates (per 1000 enrollees) at 5 years were 488.3 for patients who were given counseling compared to 579.3 for patients without counseling. After adjustment for age, race, gender, prior histories of hypertension, cardiovascular diseases, diabetes, and chronic obstructive pulmonary disease, Killip class III or IV, and discharge to a skilled nursing facility; inpatient counseling remained associated with improved survival (relative hazard, 0.78; 95% confidence interval, 0.63-0.97). CONCLUSIONS: Inpatient counseling on smoking cessation is suboptimal among older smokers hospitalized with AMI. Even without confirmation of actual cessation, these data suggest that provision of smoking cessation advice or counseling has a major impact on survival of older adults. PMID- 14751322 TI - Perceived environment attributes, residential location, and walking for particular purposes. AB - BACKGROUND: Identifying environmental factors that can influence physical activity is a public health priority. We examined associations of perceived environmental attributes with walking for four different purposes: general neighborhood walking, walking for exercise, walking for pleasure, and walking to get to and from places. METHODS: Participants (n =399; 57% women) were surveyed by mail. They reported place of residence, walking behaviors, and perceptions of neighborhood environmental attributes. RESULTS: Men with the most positive perceptions of neighborhood "aesthetics" were significantly more likely (odds ratio [OR]=7.4) to be in the highest category of neighborhood walking. Men who perceived the weather as not inhibiting their walking were much more likely (OR=4.7) to be high exercise walkers. Women who perceived the weather as not inhibiting their walking were significantly more likely to be high neighborhood walkers (OR=3.8) and those with moderate perceptions of "accessibility" were much more likely to do more walking for pleasure (OR=3.5). CONCLUSIONS: Different environmental attributes were associated with different types of walking and these differed between men and women. Approaches to increasing physical activity might usefully focus on those attributes of the local environment that might influence particular subsets of walking behavior. PMID- 14751323 TI - Body mass index and quality of well-being in a community of older adults. AB - BACKGROUND: The impact of obesity and associated conditions on health has not been assessed in older adults using a generic, utility-based measure of health related quality of life (HRQOL). This study evaluates the relationship between body mass index (BMI) and HRQOL scores and gives estimates of quality-adjusted life years (QALYs) lost to overweight, obesity, and associated conditions. METHODS: A total of 1326 adults from the Rancho Bernardo longitudinal cohort study, with a mean age of 72 years, completed the Quality of Well-Being Scale (QWB), a generic health-related quality of life measure. Height, weight, exercise, and smoking status were also assessed. Differences in QWB scores between obese adults and those with a normal BMI were used to estimate the QALYs lost due to obesity and associated conditions. RESULTS: Participants were divided into four groups based on BMI: <20 (underweight); 20 to 24.9 (normal); 25 to 29.9 (overweight); >30 kg/m(2) (obese). Analysis of covariance controlling for age, gender, smoking history, and exercise showed a significant difference between group means (F(7,1310)=30.79; p <0.001). The normal BMI group had the highest QWB score (0.709), followed by the underweight (0.698), overweight (0.695), and obese (0.663) groups. The QWB score for the obese group was significantly lower than that for the normal and overweight groups. An estimated 2.93 million QALYs are lost in this country each year from obesity and associated conditions. CONCLUSIONS: Obese older adults tend to have lower HRQOL than those who are overweight or of normal BMI. The lower QWB scores associated with obesity translate into millions of QALYs lost each year. Being overweight but not obese did not have a significant impact on HRQOL in this population. PMID- 14751324 TI - Content analysis of prime-time television coverage of physical activity, 1970 2001. AB - BACKGROUND: The purpose of this study was to examine how major nightly television networks reported on the health benefits of physical activity. METHODS: A retrospective content analysis of physical activity coverage on four major nightly television networks from 1970 to 2001 was performed. The Vanderbilt Television News Archives were searched for keywords "physical activity," "physical fitness," and "exercise." RESULTS: During the 31-year time period, 111 non-overlapping reports aired on all networks combined. The link between physical activity and health was reported in 53 (47.7%) articles, with general health (n =16, 14.4%) and heart disease (n =12, 12.6%) cited most frequently. Just three broadcasts related to the Surgeon's General Report on Physical Activity and Health were aired following its publication in 1996. CONCLUSIONS: Although the protective health benefits of physical activity are well established, physical activity received a modest amount of television coverage from 1970 through 2001. PMID- 14751325 TI - Risk perceptions regarding ticks and Lyme disease: a national survey. AB - BACKGROUND: Lyme disease (LD) is caused by the tickborne bacterium Borrelia burgdorferi and, in 2000, accounted for >90% of all reported cases of vectorborne illness in the United States. Aside from anecdotal and indirect evidence, little empirical evidence exists regarding what the U.S. public knows, says, or does about preventing LD. OBJECTIVES: To examine knowledge, perceptions, and practices regarding prevention of tick bites and LD. METHODS: In 1998, a random-digit-dial frame was used to collect a cross-sectional sample (n =1500) from the 48 coterminous states plus the District of Columbia, and an over-sample (n =250) from six states with the highest incidence of LD. RESULTS: Forty percent of respondents reported doing something to avoid being bitten by ticks. Less than half (41%) used insect repellent. Ninety-two percent of those who had heard about LD stated their likelihood of ever getting the disease was 51% v >17.6%, p =0.001). CONCLUSIONS: The results suggest that a brief nurse-delivered intervention is effective at increasing patient adherence with SSE. Utilizing digital photographs as an adjunct to screening appeared to increase patient adherence to performing SSE. PMID- 14751329 TI - Do the majority of Asian-American and African-American smokers start as adults? AB - BACKGROUND: Identifying ethnic differences in the age of smoking onset from nationally representative data can lead to improved targeted prevention programs and policies to combat smoking in ethnic communities. METHODS: Analyzing data from the Tobacco Use Supplements of the U.S. Census Bureau's Current Population Surveys throughout the 1990s, differences in the age of regular smoking onset among Asians/Pacific Islanders (A/PI), African Americans (AA), Hispanics/Latinos (H/L) and non-Hispanic whites (WH) are reported. Data on people aged 26 to 50 years at the time of the survey interview (n =130,356; mean age=38.4 years; 47.9% male; 1.9% A/PI, 7.8% AA, 5.2% H/L, and 85.1% WH) were examined. RESULTS: Results indicate significant ethnic disparities in when people start smoking, among A/PIs in particular, and AAs and H/Ls to a lesser degree, who initiate regular smoking at later ages than do WHs. The majority of A/PIs and AAs initiated smoking as young adults, with almost half (47.8%) of A/PIs who were ever regular smokers starting between ages 18 and 21, compared with 39.8% of AAs, 37.5% of H/Ls, and 36.7% of WHs. CONCLUSIONS: These findings indicate significant ethnic disparities in relation to when people start smoking, with the majority of A/PIs and AAs initiating as young adults. The findings suggest that prevention strategies should begin at a young age and continue throughout young adulthood, especially among ethnic minority populations. Further consideration of the different influences on later initiation in ethnic minorities may lead to suggestions to improve current smoking-prevention programs aimed at adolescents and young adults. PMID- 14751330 TI - Social determinants of tuberculosis case rates in the United States. AB - BACKGROUND: Social capital has been related to a number of important public health variables such as overall mortality, health status, and sexually transmitted infections (including AIDS case rates). However, the relationship of social capital to tuberculosis has received little attention. Because social capital may be related to the constructs of poverty and income inequality, any exploration of the correlation between social capital and tuberculosis should include examination of the interrelationships with poverty and income inequality as well. OBJECTIVE: This study examined the state-level relationship between social capital, poverty, income inequality, and tuberculosis case rates. METHODS: The design was state-level, correlational analysis (including bivariate linear correlational analysis, and multivariate linear stepwise regression analysis). Main outcome measures were 1999 state-level case rates of tuberculosis. RESULTS: In bivariate analyses, poverty, income inequality, and social capital were all significantly correlated with tuberculosis case rates. In stepwise multiple regression analyses predicting tuberculosis case rates from this set of three predictor variables, social capital and income inequality entered the regression equation (with social capital being the strongest predictor variable). CONCLUSIONS: These results suggest that social capital is highly predictive of tuberculosis at the state level. The results indicate the need for further research into this potentially causal relationship, including the examination of structural interventions designed to increase social capital. PMID- 14751331 TI - Tuberculosis prevention in Mexican immigrants: limitations of short-course therapy. AB - BACKGROUND: Two months of rifampin and pyrazinamide (RIF/PZA) for tuberculosis prevention has been advocated as a way to improve adherence in mobile populations, such as recent immigrants. However, RIF/PZA requires intensive patient and laboratory monitoring for hepatotoxicity. OBJECTIVES: To describe the feasibility and outcomes of using RIF/PZA for TB prevention during a tuberculosis outbreak in a Mexican immigrant community, where 23 adults and 11 children were treated with RIF/PZA between August 2001 and October 2001. METHODS: Retrospective chart review and interviews with health department employees were conducted to assess completion rates, hepatotoxicity, cost, and feasibility of monitoring. RESULTS: Ten (91%) children and 13 (57%) adults completed RIF/PZA. One child (9%) and four adults (17%) developed drug-induced hepatitis. Cultural barriers affected care. The adults resisted the biweekly blood draw, believing it would "drain them of energy." RIF/PZA, plus monitoring, was twice as costly as 4 months of rifampin. CONCLUSIONS: RIF/PZA was associated with significant hepatotoxicity, poor completion, and cultural barriers to monitoring, and was more costly than standard therapy. Tuberculosis prevention must address potential clinical, cultural, and economic barriers to completion and monitoring of short-course therapy in immigrants. PMID- 14751332 TI - The tobacco industry's political efforts to derail the EPA report on ETS. AB - Previously secret tobacco industry documents detailed a multifaceted approach of political strategies aimed to derail the 1993 Environmental Protection Agency (EPA) risk assessment on environmental tobacco smoke (ETS). These pervasive strategies included the following: (1) lobbying the first Bush Administration to approve an executive order that would impose new risk assessment standards for federal agencies, thus delaying the release of the EPA report; (2) having the first Bush Administration transfer jurisdiction over ETS from the EPA to Occupational Safety and Health Administration (OSHA), thus obviating the need for the release of the EPA report; and (3) applying enormous political pressure directly by alleging improper procedure and policy at EPA. Although some of the attempted strategies failed, the political pressure from Congressman Thomas Bliley (R-VA) was a success. This is the first report showing how a single member of Congress in conjunction with his staff, tobacco industry attorneys, and executives worked very aggressively to do the tobacco industry's bidding. These tactics successfully delayed the EPA risk assessment and placed a cloud over its validity that was not fully vindicated until December 2002 when the U.S. 4th Circuit Court of Appeals overturned the tobacco industry's suit against the EPA. The documents show that the industry will expend whatever effort is necessary to protect itself from public health policy that would adversely affect consumption of cigarettes and, therefore, profit. PMID- 14751333 TI - Considering schizophrenia from a prevention perspective. AB - Schizophrenia is a serious mental illness that causes major disability and psychosocial impairment. Recent advances in the neurosciences are prompting considerations of schizophrenia from a preventive perspective. An overview of the literature is provided on two important aspects of the development of a prevention orientation in schizophrenia research: elucidation of potential causal risk factors for schizophrenia and research on risk markers. Risk factors for schizophrenia include, but are not limited to, family history, older paternal age, velo-cardio-facial syndrome, maternal infections during pregnancy, pregnancy and delivery complications, and social adjustment difficulties in childhood and adolescence. Potential risk markers include structural brain pathology, minor physical anomalies and dermatoglyphic abnormalities, neurocognitive deficits, eye tracking dysfunction, certain electrophysiologic findings, and olfactory identification deficits. Several early efforts at indicated preventive interventions targeting individuals at particularly high risk for developing the disorder are discussed. The preventive medicine and public health disciplines may have a role in future research and interventions that apply a preventive perspective to schizophrenia and other mental illnesses. Like any other chronic medical condition, schizophrenia can be considered from a preventive perspective. PMID- 14751334 TI - Response to the 2003, vol. 23 no. 3s1 poem entitled "ain' like there's hunger". Move insecurity. PMID- 14751335 TI - Assisted reproductive technology in the new millennium: part I. PMID- 14751336 TI - Changes in urinary parameters after oral administration of potassium-sodium citrate and magnesium oxide to prevent urolithiasis. AB - OBJECTIVES: To examine urinary parameters among normal individuals and patients with calcium oxalate (CaOx) stones after oral administration of potassium-sodium citrate (KNa-Cit) and magnesium oxide (MgO). Urinary citrate and magnesium have been known as the inhibitors of CaOx stone formation. Supplementation with potassium-magnesium citrate prevents the recurrence of CaOx stones. METHODS: Twenty-five male volunteers aged 21 to 42 years without a history of urinary stones were given either KNa-Cit or MgO, or both. Fourteen patients with recurrent CaOx stones were also given both supplements, and 24-hour urine samples were collected to determine the urinary parameters. RESULTS: The administration of both KNa-Cit and MgO to the normal individuals increased the excretion of citrate, magnesium, and potassium by 70.0%, 44.2%, and 50.0%, respectively. These parameters increased less when KNa-Cit or MgO was administered individually. After administration of both supplements to the patients with stones, the citrate, magnesium, and potassium levels increased by 62.1%, 63.3%, and 25.3%, respectively, and oxalate decreased by 66.5%. In both normal individuals and patients, the ion activity product index of CaOx decreased significantly more after administration of the combination than with either compound alone or before administration. CONCLUSION: The combination of KNa-Cit and MgO is more effective than either supplement alone in inhibiting the crystallization of CaOx stones. The combination may improve the urinary parameters of patients with stones accompanied by hypocitraturia and/or hypomagnesuria. PMID- 14751338 TI - Role of mepartricin in category III chronic nonbacterial prostatitis/chronic pelvic pain syndrome: a randomized prospective placebo-controlled trial. AB - OBJECTIVES: To verify the efficacy of mepartricin versus placebo with regard to symptom improvement in patients with chronic nonbacterial prostatitis/chronic pelvic pain syndrome (CPPS) and to verify a relation between hormonal levels and clinical improvement in these patients. METHODS: Twenty-six patients with CPPS were included in our study and randomized into two groups of 13 subjects each. Group 1 patients were treated with mepartricin (40 mg daily) and group 2 patients with placebo. All patients underwent treatment for 60 days. At the beginning and end of therapy, all patients underwent evaluation, including a standardized history, physical examination, luteinizing hormone, follicle-stimulating hormone, testosterone, and beta-estradiol measurements, and a National Institutes of Health-Chronic Prostatitis Symptom Index (NIH-CPSI) questionnaire. RESULTS: We observed a decrease in the total NIH-CPSI score from 25.0 to 10.0 in group 1 and from 25.0 to 20.0 in group 2, revealing a 60% and 20% improvement in groups 1 and 2, respectively. A statistically significant decrease was observed with regard to pain (from 11.0 to 4.0 and from 10.0 to 8.0, respectively) and quality of life (from 10.0 to 5.0 and 10.0 to 9.0, respectively). No statistically significant difference was observed in urinary dysfunctions. The luteinizing hormone, follicle-stimulating hormone, and testosterone values were similar in both groups before and after treatment; the 17-beta-estradiol levels were significantly lower in group 1 compared with group 2 at the end of the study. CONCLUSIONS: Mepartricin provides significant symptomatic improvement in men with CPPS compared with placebo. The role of mepartricin in decreasing estrogen plasmatic levels and their concentration in the prostate may account for this clinical improvement. PMID- 14751339 TI - Prevalence of interstitial cystitis in first-degree relatives of patients with interstitial cystitis. AB - OBJECTIVES: To compare, in a pilot study, the prevalence of interstitial cystitis (IC) among first-degree relatives of patients with IC with the prevalence of IC in the general population. Often the first evidence that a disease may have a genetic susceptibility is the demonstration of family aggregation of the disease. METHODS: Members of the Interstitial Cystitis Association (ICA) were mailed a survey inquiring about the prevalence of the disease or consistent symptoms in first-degree family members (parents, siblings, and/or children). The same survey instrument was used in telephone interviews of a randomly selected sample of nonrespondents to determine the degree of responder bias. RESULTS: Of 2581 respondents to the mail-in survey, 101 (3.9%) reported 107 first-degree relatives with IC. The subsequent telephone interviews with 346 randomly selected nonrespondents revealed little selection bias in the mail-in survey. These measurements, plus data-based assumptions of proportions of those self-reporting IC who actually met the hydrodistension requirements for the diagnosis of IC, suggest that women, 31 to 73 years old who were first-degree relatives of patients with IC, themselves had a prevalence of IC of 995/100,000. A comparison of this with the number approximating the prevalence in the general population of American women of this age (60/100,000) indicates a risk ratio for IC in adult female first-degree relatives of 17. CONCLUSIONS: Adult female first-degree relatives of patients with IC may have a prevalence of IC 17 times that found in the general population. This, together with previously reported evidence showing a greater concordance of IC among monozygotic than dizygotic twins, suggests, but does not prove, a genetic susceptibility to IC. PMID- 14751340 TI - Antiproliferative factor, heparin-binding epidermal growth factor-like growth factor, and epidermal growth factor in men with interstitial cystitis versus chronic pelvic pain syndrome. AB - OBJECTIVES: To determine whether men with chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) have urine markers previously described for patients with interstitial cystitis (IC; presence of antiproliferative factor [APF] activity, decreased levels of heparin-binding epidermal growth factor-like growth factor [HB-EGF], and increased levels of epidermal growth factor). METHODS: Clean catch urine specimens were collected from 41 symptomatic patients with CP/CPPS, 36 asymptomatic men without bladder disease who served as the control group, and 24 men with IC. APF activity was determined by (3)H-thymidine incorporation into primary normal adult human bladder epithelial cells. HB-EGF and epidermal growth factor levels were determined by enzyme-linked immunosorbent assay. RESULTS: Men with CP/CPPS did not differ significantly from asymptomatic controls for any of the three markers tested (P >0.49). In contrast, APF activity was present significantly more often and HB-EGF levels were significantly lower in the urine specimens from men with IC than in the specimens from controls or patients with CP/CPPS (P <0.00001 for all four comparisons). Although the epidermal growth factor levels also tended to be higher in the urine from patients with IC than in the urine from controls, the difference did not reach statistical significance (P = 0.06). CONCLUSIONS: These findings indicate that at least two of the urine biomarkers previously identified in women with IC (presence of APF activity and decreased levels of HB-EGF) are also found in men with IC, but not in men with CP/CPPS. This finding suggests that IC and CP/CPPS may be two different disorders with distinct pathophysiologies. It also confirms the utility of the presence of APF activity and HB-EGF levels as markers for IC in men, as well as in women, with this disorder. PMID- 14751341 TI - Complications of abdominal urologic laparoscopy: longitudinal five-year analysis. AB - OBJECTIVES: To analyze complications of abdominal laparoscopic surgery of the urinary tract at a single institution during a 5-year period. METHODS: From 1996 to 2000, we identified 894 abdominal laparoscopic procedures performed at a single institution: 600 nephrectomies (live donor, simple, radical, nephroureterectomy, and partial), 112 pyeloplasties, 61 renal biopsies, 35 retroperitoneal lymph node dissections, 31 renal cyst ablations, 18 adrenalectomies, and 37 other abdominal procedures. The charts were retrospectively reviewed for complications, which were classified as operative, postoperative, or medical. Complications were correlated with patient age and American Society of Anesthesiologists score. Statistical analysis was performed with Fisher's exact test and chi-square tests. RESULTS: A total of 118 complications (13.2%) occurred. Two patients (0.2%) died. As a result of operative complications, the procedure of 13 patients (1.5%) was converted to an open one. As a result of postoperative complications, 13 (1.5%) underwent operative and 6 (0.7%) nonoperative intervention. The most common intraoperative complications were vascular (n = 23), adjacent organ (n = 10), and bowel (n = 9) injuries. The most common postoperative complications were neuromuscular pain (n = 12), hematoma (n = 11), urine leak (n = 7), and wound infection (n = 7). The differences in the annual complication rates for all procedures did not attain statistical significance (P = 0.5). Among all procedures, excluding live donor nephrectomy, complications of any kind correlated with a greater patient American Society of Anesthesiologists score (P = 0.01). CONCLUSIONS: Rather than decreasing, the overall incidence of laparoscopic complications did not change significantly during a 5-year period at our institution. The factors contributing to this observation likely included the progression of inexperienced individual surgeons through the learning curve, the introduction of new, more sophisticated laparoscopic procedures, and stable rates of patient comorbidity. This experience may represent the average complication rate for urologic laparoscopy at a large volume, academic training center. PMID- 14751342 TI - Role of voided urine cytology in diagnosing primary urethral carcinoma. AB - OBJECTIVES: To evaluate the role of voided urine cytology in diagnosing primary carcinoma of the urethra in male and female patients. METHODS: We reviewed the medical records of all patients with urethral carcinoma seen at Memorial Sloan Kettering Cancer Center between 1958 and 1996. The patients who had undergone voided urine cytology before any treatment were the subject of this report. RESULTS: This report included 41 female and 29 male patients. In the cohort of female patients, the most common histologic type was adenocarcinoma (n = 16), followed by squamous cell carcinoma (SCC; n = 9) and transitional cell carcinoma (TCC; n = 6). Urine cytology was positive in 24 patients (59%). The sensitivity was greatest in patients with SCC (77%) and lowest in patients with TCC (50%). In the cohort of male patients, the most common histologic type was SCC (n = 14) followed by TCC (n = 10). Urine cytology was positive in 16 patients (55%). The sensitivity was greatest for patients with TCC (80%) and lowest for patients with SCC (50%). CONCLUSIONS: Voided urine cytology is not a very reliable method of diagnosing primary carcinoma of the urethra in either male or female patients. A cystoscopic evaluation with possible biopsy is warranted if suspicion of carcinoma of the urethra is high. PMID- 14751343 TI - Laparoscopic versus open donor nephrectomy: ureteral complications in recipients. AB - OBJECTIVES: To describe our experience with laparoscopic donor nephrectomy (LDN) and open donor nephrectomy (ODN) regarding ureteral complications. LDN has proved to be safe and to offer significant benefits to the donor compared with ODN. Of major importance is the effect of the surgical technique on the graft. Studies have shown an increased incidence of ureteral complications in recipients of laparoscopically procured kidneys. Operative reconstruction results in additional morbidity for the recipient. METHODS: Living donors and their recipients, who underwent surgery from January 1994 to April 2002, were included in this retrospective study. A total of 122 LDN and 77 ODN recipients were included. RESULTS: Of the 122 LDN and 77 ODN recipients, 15 (12%) and 10 (13%), respectively, required percutaneous nephrostomy drainage. In total, 5 LDN (4.1%) and 5 ODN (6.5%) recipients required reconstruction of the ureter because of obstruction of the ureter or urine leakage (P value not statistically significant, excluding reconstruction required for technical errors). The operating time, warm ischemia time, and serum creatinine were comparable between recipients with or without ureter complications requiring reconstruction. CONCLUSIONS: In our experience, LDN was not associated with an increased incidence of ureteral complications in the recipient compared with ODN. PMID- 14751345 TI - Natural history and clinical outcome of sporadic renal cortical tumors diagnosed in the young adult. AB - OBJECTIVES: To determine the natural history of patients younger than 40 years (young patient [YP] group) who are diagnosed with a sporadic renal cortical tumor (RCT) and to compare the natural history of these patients with the more typical older patient (OP) with RCT. METHODS: We reviewed our database and identified 34 patients (younger than 40 years old, median age 35) who underwent surgery for a sporadic RCT. The YP group outcomes were compared with 100 patients between 41 and 85 years (median 65). We fit a Cox proportional hazards model to examine the relationship between age at presentation and recurrence risk. RESULTS: The median tumor size in the YP group was 3.8 cm (range 0.6 to 19) and in the OP group was 5.0 cm (range 0.9 to 22; P = 0.225). Tumors were discovered incidentally in 51% and 56% of the YP and OP groups, respectively (P = 0.65). The frequency of partial nephrectomy did not differ between the two groups (35% YP and 30% OP, P = 0.55). The frequency of malignant histologic subtypes did not differ between the groups (P = 0.439). In the YP group, only larger tumor size (hazard ratio 1.23, 95% confidence interval 1.02 to 1.50, P = 0.034) was associated with a statistically significant increased risk of recurrence. Those in the YP group were not more or less likely to develop recurrence than those in the OP group (hazard ratio 0.79, 95% confidence interval 0.22 to 2.85, P = 0.72). The 5-year disease-free survival rate was 73% and 80% in the YP and OP groups, respectively (P = 0.23). The 5-year disease-specific survival rate was 85% and 84% in the YP and OP groups, respectively (P = 0.88). CONCLUSIONS: The findings of our study indicate that the natural history of RCTs is similar in both younger and older patients. Young patients were neither more nor less likely to develop recurrence compared with their older counterparts. PMID- 14751346 TI - Prognostic value of body mass index in patients undergoing nephrectomy for localized renal tumors. AB - OBJECTIVES: To identify whether the body mass index (BMI) has any adverse effect on the prognosis of patients with established renal cell carcinoma, given the increasing prevalence of obesity and the rising incidence of renal cell carcinoma in the United States. METHODS: We reviewed the records of patients who underwent nephrectomy for localized disease between 1985 and 1998 at our institution. Patients were grouped according to BMI as normal (less than 25 kg/m2), overweight (25 to 30 kg/m2), or obese (more than 30 kg/m2). Cox regression analysis was used to determine the significant predictors of metastasis and survival. RESULTS: A total of 400 patients met the inclusion criteria. On univariate analysis, normal BMI (P = 0.018), pathologic stage (P <0.0001), Fuhrman grade (P = 0.007), maximal tumor size (P = 0.005), and time to recurrence (P <0.0001) were significant predictors of disease-specific death. On multivariate regression analysis, the time to metastasis (P <0.0001) was the best predictor of disease-specific death, followed by BMI (normal versus overweight or obese; P = 0.006) and pathologic stage (P = 0.007). Patients with a normal BMI who had pathologic Stage pT3 or greater and developed metastasis within 19 months of surgery had the worst prognosis, with a disease-specific death rate of 52.0%. CONCLUSIONS: Our findings suggest that overweight and obese patients with renal cell carcinoma have a more favorable prognosis than patients with a normal BMI. If others confirm our finding that a high BMI confers a survival advantage to patients undergoing nephrectomy, BMI may prove to be an important prognostic factor in renal cell carcinoma. PMID- 14751347 TI - Feasibility of robot-assisted totally intracorporeal laparoscopic ileal conduit urinary diversion: initial results of a single institutional pilot study. AB - OBJECTIVES: To explore the use of the da Vinci Surgical Robotic System (DSRS) to assist in the completion of totally intracorporeal laparoscopic ileal conduit urinary diversion (TLIC). METHODS: Two patients with radiation cystitis underwent TLIC procedures and another patient with bladder cancer underwent TLIC along with laparoscopic radical cystoprostatectomy at our institution. The ileal conduit urinary diversion was done totally intracorporeally using conventional laparoscopic techniques, and the DSRS was used to assist in the Bricker-type ureteroileal anastomosis. RESULTS: The 3 patients in the study included 2 men and 1 woman (mean age 73 years, range 64 to 84). The TLIC was completed intracorporeally in all 3 patients without the need for open conversion. The operative time, estimated blood loss, intraoperative decrease in hemoglobin, and time to hospital discharge for the 2 patients undergoing TLIC and the patient undergoing TLIC along with radical cystoprostatectomy was 628, 616, and 828 minutes, 50, 200, and 500 mL, 1.7, 2.8, and 5.3 g, and 5, 7, and 10 days, respectively. The median follow-up was 4.5 months (range 3.5 to 5.5). Postoperative satisfactory drainage of both kidneys was confirmed in all 3 patients at 8 weeks or later by intravenous urography or renal nuclear imaging. The serum creatinine remained stable in all 3 patients after surgery at hospital discharge. The only complication noted was postoperative ileus in the patient undergoing radical cystoprostatectomy that resolved with conservative management. CONCLUSIONS: TLIC is technically feasible and safe and can be done intracorporeally without complications. The DSRS can be successfully used to assist in the completion of TLIC. However, that each case lasted for more than 600 minutes highlights the need for further refinement in the technique. The practical application of TLIC requires improved long-term outcomes compared with open surgery, as well as a reduction in the operative time to justify the costs of robotic surgery. PMID- 14751348 TI - Applicability and reproducibility of condom catheter method for measuring isovolumetric bladder pressure. AB - OBJECTIVES: To report on the applicability, reproducibility, and adverse events of the noninvasive condom catheter method in the first 730 subjects of a longitudinal survey of changes in urinary bladder contractility secondary to benign prostatic hyperplasia, in which 1300 men will be evaluated three times in 5 years using this method. METHODS: Subjects were recruited by general practitioners, general publicity, and e-mail. Only those meeting the study criteria were entered in the study. If the free flow rate exceeded 5.4 mL/s, at least two consecutive condom pressure measurements were attempted using the condom catheter method. The condom pressure measured reflected the isovolumetric bladder pressure, a measure of urinary bladder contractility. The reproducibility of the method was quantified by a difference plot of the two maximal condom pressures measured in each subject. RESULTS: In 618 (94%) of 659 eligible participants, one condom pressure measurement was completed; two measurements were done in 555 (84%). The maximal condom pressure ranged from 28 to 228 cm H2O (overall mean 101, SD 34). A difference between the two pressures of less than +/ 21 cm H2O was found in 80%. The mean difference was -1 cm H2O (SD 18), significantly different from 0. Some adverse events such as terminal self limiting hematuria were encountered. CONCLUSIONS: The condom catheter method is very suitable for large-scale use. It has a success rate of 94% and a reproducibility comparable to that of invasive pressure flow studies. PMID- 14751349 TI - Single-blind, randomized trial of pelvic floor muscle training, biofeedback assisted pelvic floor muscle training, and electrical stimulation in the management of overactive bladder. AB - OBJECTIVES: To compare the efficacy of pelvic floor muscle training (PFMT), biofeedback-assisted PFMT (BAPFMT), and electrical stimulation (ES) in the management of overactive bladder (OAB). METHODS: The interventions for the 12 week treatment period, conducted by the physiotherapist who was unaware of the progress and outcome, included (a) a PFMT program tailored to the subject's PERFECT (power, endurance, repetitions, and fast [1-second] contractions, with every contraction timed) scheme, used for training at home; (b) an electromyography BAPFMT program and home program tailored to the subject's PERFECT scheme; and (c) an ES program using biphasic symmetric probe current with 10-Hz frequency, 400-micros pulse width, 10/5 duty cycle, and varying intensity. Identical preintervention and postintervention assessment included King's Health Questionnaire, as well as outcomes of urge incontinence and other urinary symptoms. RESULTS: Of the 103 women who completed this study, 34 were in the PFMT group, 34 in the BAPFMT group, and 35 in the ES group. The changes in the three parameters of King's Health Questionnaire revealed statistically significant differences, except for the total score, between ES and BAPFMT (domain 7, P = 0.003; domain 9, P = 0.029; and total score, P = 0.952). These same parameters were significantly different between ES and PFMT (domain 7, P = 0.007; domain 9, P = 0.001; and total score P = 0.004). The change in total score was significantly different between BAPFMT and PFMT (P = 0.003). The subjective improvement/cure rate of OAB was 51.4% for ES, 50.0% for BAPFMT, and 38.2% for PFMT (P = 0.567). CONCLUSIONS: ES had the greatest subjective reduction rate of OAB and was the most effective of the three treatments. BAPFMT was more effective than PFMT. PMID- 14751350 TI - Do objective urodynamic or clinical findings determine impact of urinary incontinence or its treatment on quality of life? AB - OBJECTIVES: To correlate the clinical and urodynamic parameters with two measures of incontinence-specific quality of life (QOL), to describe the changes in those measures after treatment, and to explore the factors determining these changes. METHODS: A total of 82 incontinent women (mean age 52 years, range 28 to 80) underwent urodynamics testing and a 48-hour pad test. They also completed the frequency/volume chart, estimated the degree of bother from urinary incontinence using the visual analog scale (VAS), and completed a validated QOL instrument- the Urinary Incontinence Severity Score (UISS). Sixty-nine women were re evaluated 13 months (range 6 to 21) after treatment. RESULTS: A greater degree of disability from urinary incontinence as measured by the VAS correlated with a lower maximal urethral closure pressure (r = -0.29, P <0.01), greater detrusor pressure (r = 0.30, P <0.05), and amount of urine leakage (r = 0.46, P <0.001). The UISS correlated poorly with the urodynamic and frequency/volume chart findings. A greater amount of urine leakage was the best predictor of QOL impairment as measured by the UISS (beta 0.25; P = 0.034). The change in urine leakage best predicted the change in the UISS (beta 0.30; P = 0.024) and the change in the VAS (beta 0.48; P = 0.001). CONCLUSIONS: The response to the question "How bothered are you by incontinence at this moment?" best reflects the severity of urinary incontinence measured objectively. Urodynamic parameters correlated poorly with incontinence-specific QOL measure. Of the clinical objective measures of the severity of urinary incontinence, the amount of leakage in the pad test was the best, although modest, predictor of QOL impairment. The change in urine leakage best predicted the change in QOL scores and VAS 1 year after beginning treatment. PMID- 14751352 TI - Concurrent chemoradiotherapy for clinical stage T2 bladder cancer: report of a single institution. AB - OBJECTIVES: To report our experience with concurrent chemoradiotherapy for clinical Stage T2 bladder cancer. METHODS: From 1996 to 2002, 43 patients were treated with concurrent chemotherapy and radiotherapy for clinical Stage T2 bladder cancer. After complete bladder transurethral resection, all patients underwent chemotherapy, consisting of one daily infusion of cisplatin at a dose of 15 mg/m2 and 5-fluorouracil at a dose of 400 mg/m(2) on days 1 to 3 (first cycle) and days 15 to 17 (second cycle). Pelvic irradiation was administered at a dose of 24 Gy, using two daily fractions of 3 Gy on days 1, 3, 15, and 17. Random biopsies were performed 6 weeks after the end of the first two cycles. Patients with persistent invasive tumor underwent cystectomy; others received two additional cycles of concurrent chemoradiotherapy. RESULTS: The mean follow-up was 36.3 months (range 3 to 72). Nine patients underwent early cystectomy for nonresponse, and 2 patients underwent delayed cystectomy. The overall rate of cystectomy was 25.6%. The rate of specific survival at 3 and 5 years was 75% and 60%, respectively. The overall rate of recurrence-free survival at 3 and 5 years was 63% and 33%, respectively. Two factors correlated with patient survival: the presence of carcinoma in situ at first resection (P = 0.01) and the response after the first two cycles (half dose; P = 0.004). CONCLUSIONS: In our experience, concurrent chemoradiotherapy is less effective than primary cystectomy for clinical Stage T2 bladder cancer. This treatment may be unwarranted in patients with concomitant carcinoma in situ at the first resection. PMID- 14751353 TI - Geographic distribution of prostate cancer incidence in the era of PSA testing, Connecticut, 1984 to 1998. AB - OBJECTIVES: To evaluate the geographic variation of prostate cancer incidence in Connecticut during a 15-year period: before the introduction of prostate-specific antigen (PSA) testing (1984 to 1988), during its introduction (1989 to 1993), and after its widespread adoption (1994 to 1998). METHODS: A spatial scan statistic was used to detect and test the significance of the geographic rate variation among 29,944 incident cases. RESULTS: During 1984 to 1988, eight locations had incidence rates that differed significantly from the statewide level; for 1989 to 1993, only two locations were identified. By 1994 to 1998, a significant rate variation was noted for 10 locations around Connecticut. CONCLUSIONS: The observed geographic variation of incidence rates may reflect differing opportunities to uncover latent cases at given locales. PMID- 14751354 TI - Individual submission and embedding of prostate biopsies decreases rates of equivocal pathology reports. AB - OBJECTIVES: To determine whether the use of biopsy kits with 6 to 12 containers in which biopsies of the prostate are individually submitted and processed reduces the monthly rates of equivocal diagnoses. METHODS: We searched our computer records for prostate needle biopsies submitted in 1 to 2 containers between July 1998 and June 2000 (group 1, 515 patients) and biopsies submitted individually in 6 to 12 containers between January 2001 and December 2002 (group 2, 933 patients). We analyzed the patient demographics and pathologic diagnoses between the two groups, including the rates of equivocal diagnoses, which included atypical gland suspicious for adenocarcinoma (ATYP) and high-grade prostatic intraepithelial neoplasia (PIN) with adjacent ATYP. RESULTS: Group 2 had statistically significant reductions in the monthly rates of equivocal diagnosis (2.8% versus 6.0%, P = 0.003), ATYP diagnosis (1.8% versus 3.5%, P = 0.042), and PIN with adjacent ATYP diagnosis (1.0% versus 2.5%, P = 0.038). The differences in the monthly prostatic adenocarcinoma rates (36.9% versus 35.9%, P = 0.388) and high-grade PIN rates (13.5% versus 12.3%, P = 0.311) between the two study groups were not statistically significant. CONCLUSIONS: Multiple needle biopsies submitted in 1 to 2 containers tend to entangle and fragment and are difficult to embed in a single plane during processing. The resulting loss of tissue surface area makes a definitive diagnosis difficult on small foci of atypical glands, resulting in equivocal pathology reports. The results of our study indicate that individual submission and processing of prostate biopsies in 6 to 12 container kits reduces the monthly rates of equivocal diagnoses. PMID- 14751355 TI - Parasagittal biopsies add minimal information in repeat saturation prostate biopsy. AB - OBJECTIVES: To compare the outcome and efficacy of lateral biopsies with parasagittal biopsies in detecting prostate cancer during repeated biopsies performed using the "saturation" technique, which includes 24 cores per biopsy. Prostate biopsy may miss cancer in up to 38% of men eventually found to harbor the disease. Lateral biopsies are more likely than parasagittal biopsies to detect adenocarcinoma according to the findings of several studies. METHODS: A total of 100 patients, average age 62.1 +/- 7.9 years, underwent repeated transrectal ultrasound-guided saturation biopsy. The study group included 31 patients with previous biopsy results demonstrating high-grade prostatic intraepithelial neoplasia, 7 with atypia, and 62 with benign prostatic tissue but persistently elevated prostate-specific antigen levels. Patients had undergone an average of 1.65 previous biopsies. The average prostate-specific antigen level was 9.4 +/- 6.8 ng/mL. Biopsies were obtained from five sectors on each side and examined histologically. RESULTS: Cancer was detected in 25 (25%) of the 100 patients. Malignancy was identified in the lateral cores of all patients with positive biopsies. Parasagittal biopsy cores were positive in association with a lateral-based biopsy in 9 (36%) of the 25 malignancies, for an overall parasagittal biopsy core rate of 9% (9 of 100 patients). No cancers were detected in the parasagittal biopsy cores alone. CONCLUSIONS: Inclusion of parasagittal zone biopsy cores proved to have a low yield in detecting cancer on repeated biopsy. As all patients found to have cancer in the parasagittal biopsy cores also had cancer on the lateral biopsy cores, most time and effort can be spent obtaining lateral biopsy cores to increase the sensitivity on repeated saturation biopsy. PMID- 14751356 TI - Low AUA symptom score independently predicts positive prostate needle biopsy: results from a racially diverse series of 411 patients. AB - OBJECTIVES: To evaluate the prebiopsy parameters, including the American Urological Association symptom score (AUASS), that may be predictive of positive biopsy. Transrectal ultrasound (TRUS) biopsy of the prostate represents the reference standard in the diagnosis of prostate cancer. METHODS: A total of 411 consecutive men undergoing TRUS biopsy were prospectively evaluated. The indications for biopsy were abnormal digital rectal examination (DRE) findings and/or an elevated prostate-specific antigen (PSA) level. A single surgeon (C.R.P.) examined all the men. DRE and TRUS were each given a level of suspicion between 1 (low suspicion--smooth DRE, homogeneous TRUS) and 5 (high suspicion- hard DRE, hypoechoic lesion). A level of suspicion of 3 or greater was considered abnormal. The prebiopsy parameters examined included PSA level, age, race, biopsy history, prostate volume, TRUS-detected lesion, and AUASS. RESULTS: Of 411 men, 62% were African American and 38% were white. The mean PSA level was 11.6 ng/mL. The mean patient age was 65.3 years. Overall, 39% of men had abnormal DRE and 32% abnormal TRUS findings. The mean AUASS was 9.3. The positive biopsy rate was 40.8%. Univariate analysis demonstrated that age, PSA level, prostate volume, abnormal DRE findings, TRUS-detected lesion, and AUASS (less than 7, low) were all predictive of a positive biopsy (P <0.05). Race was not statistically significant (P = 0.38). Detailed analysis of the AUASS in the 411 men indicated that 41% had low symptom scores (less than 7), 32% had moderate scores (8 to 19), and 27% had severe scores (20 to 35). In the group of men with low symptom scores (n = 169), univariate analysis demonstrated that age, PSA level, prostate volume, and abnormal TRUS findings were all statistically significant predictors of positive biopsy (P <0.05). Multivariate analysis of the data from the 411 men demonstrated that age, PSA level, prostate volume, abnormal DRE findings, and low AUASS were all independent predictors of positive biopsy (P <0.05). CONCLUSIONS: In this prospective study, the independent predictors of positive TRUS biopsy included age, PSA level, prostate volume, abnormal DRE findings, and low AUASS. A low AUASS may be an important variable to consider when counseling patients before biopsy and when designing patient algorithms for prostate biopsy. PMID- 14751357 TI - Minor modifications in apical dissection of radical retropubic prostatectomy in patients with clinical stage T2 prostate cancer reduce positive surgical margin incidence. AB - OBJECTIVES: To describe modifications in the apical dissection of radical retropubic prostatectomy (RRP) in men with palpable disease that reduce the incidence of positive apical surgical margins. Positive surgical margins occur in up to 45% of patients undergoing RRP, even in contemporary series. The apex is the most common site of positive margins, and patients with palpable (clinical Stage T2) disease are more likely to have positive surgical margins than patients with nonpalpable (clinical Stage T1c) disease. METHODS: Ninety-five men, 48 to 77 years old (mean age 59.6) with clinical Stage T2 tumors underwent RRP between October 1998 and May 2001. Surgical modifications included identification of the prostatic urethral junction by dissection of the levator ani muscle from the prostate and partial transection of the puboprostatic ligaments; variable division of the membranous urethra 1 to 3 mm distal to the prostatic urethral junction; division of prostatic ischial ligaments before dividing the catheter and posterior membranous urethra; and division of the neurovascular bundle at the urogenital diaphragm before liberating the prostate from the rectum. RESULTS: Positive surgical margins occurred in only 8 (8.4%) of 95 patients, even though tumor extended beyond the prostatic capsule in 55% of cases. Only 2 (2.1%) of the 95 patients had isolated positive apical surgical margins. Urinary continence was not adversely affected by these modifications. CONCLUSIONS: Easily applied modifications in the apical dissection of RRP significantly reduce the incidence of positive apical margins in patients with clinical Stage T2 prostate cancer. PMID- 14751358 TI - Laparoscopic radical prostatectomy: a multi-institutional study of conversion to open surgery. AB - OBJECTIVES: To perform a multi-institutional review of the incidence and factors contributing to conversion from laparoscopic radical prostatectomy (LRP) to open radical retropubic prostatectomy (RRP) among eight surgeons. METHODS: The medical records from all patients undergoing LRP at seven institutions were reviewed to assess the incidence of open conversion to RRP. The clinical, operative, pathologic, and functional outcomes of patients who required conversion were compiled and analyzed. RESULTS: Of 670 operations, 13 (1.9%) were converted from LRP to RRP. The most common steps at which conversion occurred were the apical dissection (38%) and the posterior seminal vesicle dissection (31%). Failure to progress was the most common cause of conversion, followed by injury to adjacent structures and hypercarbia. Comorbidities associated with conversion were prior pelvic surgery and obesity (body mass index greater than 30). Six of the 13 conversions occurred in the surgeons' first 5 cases. Despite open conversion, patients were continent (no pads) (92%) and potent (44%) with at least 6 months of follow-up. CONCLUSIONS: Open conversion from LRP to RRP is an uncommon event, but is more likely to occur during a surgeon's early experience. Surgeons beginning their experience with LRP are advised to select their patients carefully to minimize the need for conversion to open surgery. Obese patients and those at risk of periprostatic adhesions should initially not be considered for LRP. Despite open conversion, the functional outcomes did not appear to be adversely affected. PMID- 14751359 TI - Treatment options for localized prostate cancer: quality-adjusted life years and the effects of lead-time. AB - OBJECTIVES: The purposes of this study were to estimate the difference in quality adjusted life-years between conservative management and prostatectomy or radiotherapy (RT) by clinical Gleason score (2 to 4, 5 to 6, 7, and 8 to 10) for patients aged 55 years and older with clinically localized prostate cancer and to adjust for and explore the effects of lead-time. For localized prostate cancer, it is not known whether treatment (prostatectomy or RT) results in longer quality adjusted survival than conservative management. Observed survival benefits after treatment may be biased by the lead-time resulting from early diagnosis with prostate-specific antigen screening. METHODS: A Markov simulation was developed, and transition probabilities were derived from a review of published studies. Utility weights were measured in male volunteers older than 60 years. Estimates of disease progression during conservative management were adjusted for lead time. Sensitivity analyses were performed on all parameters (including estimates for lead-time). RESULTS: For Gleason score 2 to 4 cancer, conservative management yielded the greatest number of quality-adjusted life-years. For Gleason score 5 to 6 cancer, any of the options appeared beneficial, depending on the estimates for disease progression. For Gleason score 7 to 10 cancer, prostatectomy and RT resulted in more quality-adjusted life-years than conservative management; with a lead-time adjustment of greater than 10 years, the outcomes with conservative management and prostatectomy were similar. The choice between prostatectomy and RT was sensitive to estimates of disease progression after treatment. CONCLUSIONS: Conservative management is a reasonable option for Gleason score 2 to 4 cancer and for some patients with Gleason score 5 to 6 cancer. Prostatectomy or RT is recommended for Gleason score 7 to 10 cancer. The survival benefits after treatment were not explained by the lead-time alone. PMID- 14751360 TI - PSA bounce predicts early success in patients with permanent iodine-125 prostate implant. AB - OBJECTIVES: To determine the clinical and dosimetric factors that predict prostate-specific antigen (PSA) bounce after iodine-125 prostate brachytherapy and to determine the predictive value of PSA bounce relative to biochemical relapse-free survival (bRFS). METHODS: A multivariate analysis of factors thought to predict for PSA bounce was performed in 295 consecutive patients with T1-T2 prostate cancer treated by prostate brachytherapy as the sole radiotherapeutic modality and a minimum follow-up of 2 years. The variables examined included age, initial PSA level, biopsy Gleason score, use of androgen deprivation, occurrence of PSA bounce, dose received by 90% of the prostate gland, and volume of gland receiving 100% of the prescribed dose. A PSA bounce was defined as a rise of at least 0.2 ng/mL greater than a previous PSA level with a subsequent decline equal to, or less than, the initial nadir. A second analysis investigating the same factors and adding PSA bounce as a predictor of bRFS was also performed. RESULTS: The median follow-up was 38 months. A PSA bounce was noted in 82 (28%) of 295 patients. On multivariate analysis, only younger age (younger than 65 years) significantly predicted for a PSA bounce. Patients who experienced a PSA bounce were less likely to have biochemical failure (P = 0.037). Overall, the bRFS rate at 5 years in those experiencing a PSA bounce was 100% versus 92% in those with no bounce. CONCLUSIONS: Immediate salvage therapy in patients with a rising PSA level after permanent prostate brachytherapy should not be initiated provided the PSA increase does not exceed the pretreatment PSA value. A PSA bounce may be associated with improved bRFS but was not associated with any of the pretreatment clinical and dosimetric factors examined. PMID- 14751361 TI - Phase I and clinical pharmacology of a type I and II, 5-alpha-reductase inhibitor (LY320236) in prostate cancer: elevation of estradiol as possible mechanism of action. AB - OBJECTIVES: To study the safety, pharmacokinetics, biologic activity, and preliminary efficacy of the bispecific 5-alpha-reductase inhibitor (LY320236) in prostate cancer. METHODS: Fifty-one patients with recurrent or metastatic prostate cancer were sequentially (nonrandomly) assigned in cohorts to receive one of five single daily oral doses of LY320236 (10, 50, 150, 250, and 500 mg). Serial evaluations included serum testosterone, dihydrotestosterone, androstenediol glucuronide, estradiol, and pharmacokinetics on days 1, 29, and 57. Toxicity assessments, x-rays/scans, and blood tests, including serum prostate specific antigen (PSA) determination, were done at regular intervals. RESULTS: Overall, treatment was well tolerated, with 3 of 51 patients developing reversible grade 3-4 toxicity (one diarrhea, two elevated liver enzymes). Peak blood levels (2 to 3 hours after drug administration) were greater for doses of 150 mg or greater compared with less than 150-mg doses with slow accumulation. Serum levels of testosterone, dihydrotestosterone, and androstenediol glucuronide did not change significantly during treatment; however, a statistically significant increase occurred in serum estradiol levels in both the castration and noncastration groups. One of 26 in the noncastration group and 4 (27%) of 15 in the castration group with baseline PSA levels of 5 ng/mL or greater had a 50% or greater PSA decline for 4 weeks or longer. CONCLUSIONS: LY320236 treatment is associated with modest reversible toxicity. An elevation of estradiol levels was seen in both castration and noncastration groups, although PSA declines were primarily seen in the castration group. The absence of cardiovascular toxicity suggests that this agent may be a promising alternative to exogenous estrogens in patients with prostate cancer who demonstrate evidence of disease progression after initial androgen deprivation treatment. PMID- 14751362 TI - Combination of LHRH analog with somatostatin analog and dexamethasone versus chemotherapy in hormone-refractory prostate cancer: a randomized phase II study. AB - OBJECTIVES: To evaluate prospectively the combination of a luteinizing hormone releasing hormone analog with a somatostatin analog and dexamethasone in patients with hormone-refractory prostate cancer (HRPC) in a randomized Phase II study. HRPC presents a challenging therapeutic problem. Salvage chemotherapy is the usual approach at this stage of the disease. The combination of a luteinizing hormone-releasing hormone analog with a somatostatin analog and dexamethasone has produced objective clinical responses in HRPC. METHODS: Forty patients with HRPC were randomized to receive one of two treatments. Group 1 underwent chemotherapy (estramustine 140 mg three times daily and etoposide 100 mg orally for 21 days) and group 2 the combination of a somatostatin analog (lanreotide 30 mg intramuscularly every 14 days) and dexamethasone (4 mg tapered to 1 mg), in addition to androgen ablation by orchiectomy or a luteinizing hormone-releasing hormone analog (triptorelin 3.75 mg intramuscularly every 28 days). The clinical and prostate-specific antigen (PSA) response, overall survival, time to progression, and toxicity were compared between the two groups. RESULTS: The data of 20 patients in group 1 and 18 in group 2 were analyzed. The demographic and clinical data were similar in the two groups at study entry. A PSA response (decrease of greater than 50%) was observed in 45% of group 1 and 44% of group 2. The difference was not statistically significant. A partial clinical response was observed in 29% and 30% of groups 1 and 2, respectively. Again, the difference was not statistically significant. Changes in performance status and pain score during treatment were not significantly different in the two groups. Hematologic toxicity was more frequent in group 1 (80% of patients), and mild diabetes was more frequent in group 2 (22% of patients). The overall survival was 18.8 months in group 1 and 18 months in group 2 (not statistically significant). The time to progression was 6 versus 4 months and, in the PSA responder subgroup, it was 8 versus 7.7 months in groups 1 and 2, respectively (neither difference was statistically significant). CONCLUSIONS: The results of our randomized Phase II study indicated that the new combination treatment (luteinizing hormone-releasing hormone analog, somatostatin analog, and dexamethasone) may be equally effective as salvage chemotherapy in patients with HRPC in terms of the clinical and PSA response, overall survival, and time to progression. A larger prospective Phase III trial is required to confirm our observations. PMID- 14751363 TI - Impact of infertility on quality of life, marital adjustment, and sexual function. AB - OBJECTIVES: To evaluate the hypothesis that infertility may result in a decrease in quality of life and an increase in marital discord and sexual dysfunction. The burden of infertility is physical, psychological, emotional, and financial. METHODS: Couples seeking treatment for infertility were asked to complete standardized validated questionnaires assessing quality of life (Quality of Well Being Scale-Self Administered, version 1.04), marital adjustment (Locke-Wallace Marital Adjustment Test), and sexual function (Brief Index of Sexual Functioning for Women and International Index of Erectile Function for men). Couples seeking elective sterilization served as the control subjects. RESULTS: Eighteen infertile couples and 12 couples seeking elective sterilization participated in the study. The mean age, years together, and household income were comparable. Infertile couples had made a mean of 14.5 office visits for infertility, and 83% of couples reported feeling societal pressures to conceive. The Marital Adjustment Test scores for the women of the infertile couples were significantly lower than the scores of the controls (P = 0.01); however no difference was noted in the men. A trend toward lower quality-of-life scores was noted in women (P = 0.09) but not in the men of infertile couples. No statistically significant impact on sexual functioning in women was noted; however, the men in the infertile couples had lower total International Index of Erectile Function scores (P = 0.05) and intercourse satisfaction scores (P = 0.03). CONCLUSIONS: Women in infertile couples reported poor marital adjustment and quality of life compared with controls. Men may experience less intercourse satisfaction, perhaps because of the psychological pressure to try to conceive or because of the forced timing of intercourse around the woman's ovulatory cycle. PMID- 14751364 TI - Transcriptional levels of four Y chromosome-linked AZF genes in azoospermic men and their association with successful sperm retrieval. AB - OBJECTIVES: To investigate the transcriptional levels of four azoospermia factor genes in the testis of azoospermic men and to investigate the association between transcriptional levels and the results of sperm retrieval. METHODS: Thirty-eight azoospermic men with normal karyotype and without Y chromosome gene deletions were enrolled. The amounts of USP9Y (ubiquitin specific protease 9, Y chromosome), DBY (dead box on the Y), RBMY1 (RNA-binding motif on the Y, 1), and DAZ (deleted in azoospermia) transcripts were examined by quantitative competitive-reverse transcriptase-polymerase chain reaction. The steady-state concentrations of mRNA encoding for each gene in each testicular sample were normalized by the amounts of a housekeeping gene (glyceraldehyde phosphate dehydrogenase [GAPDH]). Differences in the transcript ratios (gene transcript/GAPDH transcript) among patients with different testicular histologic findings (normal spermatogenesis, hypospermatogenesis, maturation arrest, Sertoli cell-only syndrome) were analyzed. For each gene, the association between the transcript ratios and the results of sperm retrieval was evaluated. RESULTS: No statistically significant difference was found in the transcript ratios of USP9Y and DBY among the four histologic patient groups (P = 0.33 and P = 0.21, respectively). In contrast, statistically significant decreases were found in the transcript ratios of RBMY1 and DAZ in patients with spermatogenic failure (P = 0.0002 and P = 0.002, respectively). The transcript ratios of USP9Y and DBY were not associated with the results of sperm retrieval, and the transcript ratios of RBMY1 and DAZ revealed a positive association with successful sperm retrieval. CONCLUSIONS: The decreased transcriptional levels of RBMY1 and DAZ in patients with spermatogenic failure may reflect the generalized loss of germ cells. The transcriptional levels of RBMY1 and DAZ may have the potential of becoming useful parameters in the prediction of successful sperm retrieval. PMID- 14751366 TI - Fertility after high-dose chemotherapy for testicular cancer. AB - OBJECTIVES: To describe long-term gonadal function after high-dose chemotherapy (HDC). HDC for testicular cancer was recently developed. The evaluation of testicular function after chemotherapy for testicular cancer is an important part of overall care, especially in young patients. METHODS: Between 1994 and 2001, 27 patients underwent HDC (1250 mg/m2 carboplatin, 1500 mg/m2 etoposide, and 7.5 g/m2 ifosfamide) at Kobe University Hospital. Information on gonadal function during follow-up was available for 10 of these patients. The mean patient age +/- SD at treatment was 32.2 +/- 8.4 years. The relationships among age at treatment, semen analysis, serum hormone levels (follicle-stimulating hormone, luteinizing hormone, testosterone, prolactin, and estradiol), cumulative dose of cisplatin and carboplatin, and length of follow-up were determined. RESULTS: Spermatogenesis recovered after cessation of HDC in 5 of 10 patients. Semen analysis in these patients showed the mean sperm concentration and motility at 42.4 +/- 10.4 million/mL and 67.2% +/- 17.0%, respectively. The patients were divided into azoospermic and nonazoospermic groups. The age of the nonazoospermic and azoospermic patients was 28.2 +/- 8.7 and 36.2 +/- 6.5 years, respectively. Follicle-stimulating hormone levels in the nonazoospermic group (11.7 +/- 3.4 mIU/mL) were significantly lower than in the azoospermic group (32.8 +/- 14.4 mIU/mL; P = 0.0472). No other statistically significant difference was observed in the other hormone levels or the cumulative dose of cisplatin and carboplatin between the azoospermic and nonazoospermic groups. CONCLUSIONS: Spermatogenesis recovers after HDC in some patients. Patients should be informed that they may or may not be fertile after HDC. PMID- 14751367 TI - Use of alternative and hormonal therapies in male infertility. AB - OBJECTIVES: To examine the patterns of use of alternative and hormonal therapies in men presenting for infertility evaluation. METHODS: We administered a questionnaire on the use of alternative and hormonal therapies to 500 consecutive men presenting for infertility evaluation at our male infertility clinic. The questionnaire asked about the use of specific therapies (eg, vitamins, herbal medicine, or hormones), the monthly cost of these therapies, and whether the principal healthcare provider had been made aware of the use of therapies. RESULTS: Of the 481 men who completed the questionnaire, 147 (31%) admitted to using one or more alternative therapies. Most of the men using alternative therapies (92 of 147, 63%) were taking one or more antioxidant vitamins or minerals (ie, vitamins C, E, selenium, zinc), and 18 men admitted to using herbal medicines. Of concern, 25 men reported using agents with clear hormonal activity (testosterone, clomiphene citrate), and 6 of these men had not informed their principal healthcare provider of this. CONCLUSIONS: Our data suggest that a significant percentage ( approximately 30%) of men presenting for infertility evaluation do use alternative therapies. It is important to inquire about the use of these therapies because some of these treatments may be toxic to the gonads. PMID- 14751368 TI - Risk-adapted treatment for patients with clinical stage I nonseminomatous germ cell tumor of the testis. AB - OBJECTIVES: To evaluate whether two courses of chemotherapy after orchiectomy in patients with clinical Stage I nonseminomatous germ cell testicular tumor at high risk of relapse will spare patients additional chemotherapy or surgery. METHODS: High-risk patients had one or more of the following: preorchiectomy alpha fetoprotein level of 80 ng/dL or greater, 80% embryonal cell carcinoma or greater, or vessel invasion in the primary tumor. Low-risk patients had none of these factors or had 50% teratoma or more without vessel invasion. High-risk patients were offered two 21-day courses of outpatient chemotherapy consisting of carboplatin, etoposide, and bleomycin. Low-risk patients and high-risk patients not receiving chemotherapy were observed. RESULTS: Of 99 patients, we classified 76 as high risk and 23 as low risk of relapse. All but eight of the high-risk patients received chemotherapy. No patient who underwent chemotherapy developed relapse, although 1 patient with normal biomarkers and a late-appearing mass underwent retroperitoneal lymphadenectomy for mature teratoma. Two of the 23 low risk patients had disease relapse; both successfully underwent chemotherapy. The nonhematologic toxicity was mild in patients receiving chemotherapy, and no patient required hospitalization. The median follow-up was 38 months (range 9 to 69). CONCLUSIONS: Two courses of postorchiectomy adjuvant chemotherapy were safe and well tolerated and markedly decreased the relapse rate in high-risk patients with clinical Stage I nonseminomatous germ cell testicular tumor without additional surgery or more protracted chemotherapy. This approach may avoid potential problems with compliance and diminish the cost of scrupulous follow-up. Our results support that surveillance for carefully selected patients at a low risk of relapse is appropriate. PMID- 14751370 TI - Assessing comfort, safety, and patient satisfaction with three commonly used penile compression devices. AB - OBJECTIVES: To assess the safety, efficacy, comfort, and patient satisfaction with three penile compression devices: the Cunningham clamp, C3, and U-Tex. METHODS: The devices were tested in random order in a multiple-period, crossover study design using a Latin squares configuration. The subjects had undergone radical prostatectomy 6 months or more before the study, had no neurologic or cognitive impairment, and had not undergone radiotherapy. Baseline penile Doppler ultrasonography was followed by ultrasound scanning with each device. In random order, subjects completed a 4-hour pad test, with and without each device, and the questionnaire. RESULTS: Twelve men completed the study. The mean Mini-Mental State Examination score was 29.6 (SD 1.2, range 27 to 30). The mean urine loss at baseline was 122.8 g (SD 130.8). The mean urine loss with each device was 53.3 g (SD 65.7) with the U-Tex, 32.3 g (SD 24.3) with C3, and 17.1 g (SD 21.3) with the Cunningham clamp (P <0.05). No device had an impact on the resistive index; the C3 and U-Tex allowed good cavernosal artery flow, and the Cunningham clamp significantly lowered the distal blood flow velocity (from 12.5 to 7.3 cm/s [left systolic velocity] to 9.5 cm/s [right systolic velocity]) even at the loosest setting. The Cunningham clamp was ranked positively by 10 of 12 men; 2 of 12 men rated the C3 positively; none rated the U-Tex positively. CONCLUSIONS: The Cunningham device was the most efficacious and most acceptable to users, but also contributed to reduced systolic velocity in all men. None of the devices completely eliminated urine loss when applied at a comfortable pressure. Individualized instruction to cognitively capable men is necessary to ensure appropriate application, comfort, and fit. PMID- 14751371 TI - Circumcision in adults: effect on sexual function. AB - OBJECTIVES: To evaluate the effects of adult circumcision on sexual function in men circumcised only for religious or cosmetic reasons. METHODS: The study group consisted of 42 male patients with a median age of 22.3 years (range 19 to 28) referred for circumcision from June 2002 to January 2003. Of the 42 men, 39 desired circumcision for religious reasons. Before circumcision, their sexual performance was evaluated using the Brief Male Sexual Function Inventory (BMSFI) and ejaculatory latency time. The BMSFI evaluation and ejaculatory latency time measurements were repeated after a postoperative interval of at least 12 weeks. The scores in the five main sections of the BMSFI and the ejaculatory latency times before and after circumcision were analyzed. RESULTS: The differences in the mean BMSFI scores were not statistically significant in any of the five sections. However, the mean ejaculatory latency time was significantly longer after circumcision (P = 0.02). CONCLUSIONS: Adult circumcision does not adversely affect sexual function. The increase in the ejaculatory latency time can be considered an advantage rather than a complication. PMID- 14751372 TI - Endoscopic management of pediatric urolithiasis in a developing country. AB - OBJECTIVES: To evaluate our experience with the endoscopic management of lower and upper urinary tract stones in pediatric patients in the Republic of Yemen. METHODS: From January 1, 1993 to December 31, 1998, 290 endoscopic operations were performed on 265 pediatric patients up to 14 years of age, 173 on the upper and 117 on the lower urinary tract. Of these procedures, 138 were percutaneous nephrolithotomy, 5 were endopyelotomy combined with percutaneous nephrolithotomy, 30 were ureteral lithotripsy, and 117 were percutaneous cystolithotomy. Of the 265 patients, 244 were boys and 21 girls (male/female ratio 11.6:1, upper tract 7.4:1, lower tract 116:1) aged 8 months to 14 years (mean age 7.1 years). The 26F adult nephroscope and 9.5F semirigid ureteroscope were used. RESULTS: The overall success rate was 98.9%. Minor complications were observed in 29 patients (10.7%); severe complication did not occur. The nucleus and/or the main component of the stones was ammonium urate in 73.5% of the cases (upper tract 54%, but for those younger than 5 years, it was 75%; lower tract 93%). CONCLUSIONS: The endoscopic management of pediatric urolithiasis is a safe and effective method. To our knowledge, this is the largest reported series on the endoscopic management of pediatric urolithiasis. PMID- 14751374 TI - Laparoscopic donor nephrectomy for pediatric recipients: outcomes analysis. AB - OBJECTIVES: To present our outcomes of laparoscopic donor nephrectomy for pediatric recipients. Laparoscopic nephrectomy has become the technique of choice for live donor nephrectomy. However, limited data are available regarding the role of this technique for pediatric recipients, who may pose special challenges and considerations, and whose parents need additional information. METHODS: Since November 1999, we have performed laparoscopic nephrectomy in more than 300 consecutive patients for live donor transplantation. Of these, 20 cases were identified that involved a recipient younger than 18 years of age. The preoperative, intraoperative, and postoperative data were reviewed to analyze the outcomes of these specific cases. The data were compared with a similar group of pediatric recipients receiving kidneys procured using the traditional open technique. RESULTS: The mean donor and recipient age was 40 years (range 26 to 52) and 13 years (range 1.7 to 18), respectively, and 19 cases (95%) involved the left kidney. The mean donor and recipient operative time was 3.3 hours (range 2.5 to 5.2) and 3.9 hours (range 2.4 to 5.1), respectively. The warm and anastomotic time averaged 4 minutes (range 2 to 8) and 30 minutes (range 16 to 41), respectively. Nineteen of the grafts functioned immediately, with a mean creatinine at 24 hours of 2.0 mg/dL (range 0.3 to 6.0). At last follow-up (mean 13.6 months), the mean creatinine was 1.1 mg/dL. Ureteral stricture requiring stent placement occurred in 2 patients (10%). No statistically significant differences were noted in operative parameters, complications, or renal function compared with a contemporary cohort of 26 pediatric patients receiving organs obtained by open donor nephrectomy. CONCLUSIONS: Our early experience with laparoscopic donor nephrectomy for pediatric recipients confirmed that the technique provides quality organs with excellent function. The method yields outcomes comparable to those after traditional open donor nephrectomy and does not require modifications for the recipient operation. PMID- 14751375 TI - Extension of a shortened ureter using the in situ appendix during Indiana pouch urinary diversion. AB - A short ureteral length can preclude the use of a continent urinary diversion during urinary tract reconstruction after bladder removal. We present a surgical technique using the in situ appendix to provide length to a shortened left ureter, thus allowing formation of a continent ileocecal urinary diversion. PMID- 14751376 TI - A new approach in the management of the hydrocele with a silicone catheter. AB - INTRODUCTION: To describe a new alternative technique to overcome several disadvantages of previous techniques to repair idiopathic hydrocele. Idiopathic hydrocele, which causes scrotal enlargement and discomfort for patients, is an abnormal fluid collection between the parietal and visceral layers of tunica vaginalis of the testis. Surgical and sclerotherapy treatments have previously been used to treat this problem. TECHNICAL CONSIDERATIONS: Forty-five patients with hydrocele (aged 19 to 67 years) underwent an alternative procedure using a silicone catheter 15 to 20 cm in length in which holes had been made to enable flow of the hydrocele fluid from the parietal layer of the tunica vaginalis to the surrounding scrotal tissue. After an incision of approximately 5 mm on the scrotal wall with a scalpel knife, a cannula was pushed through the hydrocele sac with a gentle rotation until reaching the upper scrotal wall. A silicone catheter with holes was passed through the cannula to the point of the incision by way of the cannula. At the end, both tips of the catheter were sutured. During the procedure, the hydrocele fluid was emptied using an angiocatheter before the cannula was removed. CONCLUSIONS: The results of this study indicate that this procedure does not require dissection, incision, or manipulation of the scrotal contents during treatment. It also resulted in a low rate of recurrence during the 1 to 3 years of follow-up. Additionally, the procedure has minimal complications and requires a short time, only about 15 minutes. PMID- 14751377 TI - Artificial penile nodules. PMID- 14751378 TI - Congenital vesicovaginal fistula in association with hypoplastic kidney and uterus didelphys. AB - Abnormalities of the lower urinary and genital tracts commonly coexist, although congenital vesicovaginal fistula is extremely rare. We report the first case of congenital vesicovaginal fistula and hypoplastic kidney in association with uterine didelphys. Despite extensive investigation throughout childhood, a definitive diagnosis was not made until 16 years of age. We discuss some of the difficulties that might arise in diagnosing a small vesicovaginal fistula in the presence of a fusion abnormality of the genital tract and suggest how such cases might be investigated and managed surgically. PMID- 14751379 TI - Inguinal bladder hernia masking bowel ischemia. AB - Displacement of the bladder outside of the pelvis by way of the inguinal canal represents an infrequent presentation of an inguinal hernia that rarely causes urinary tract obstruction. However, bladder hernias can become bothersome, painful, and potentially dangerous. We report a 56-year-old man with a chronic bladder hernia who became acutely unable to empty his bladder from its position in the right hemiscrotum after a motor vehicle accident. At hernia repair, bladder and ischemic bowel were discovered, mandating significant small bowel resection. This case presents interesting considerations and potential pitfalls when evaluating a patient with a bladder hernia. PMID- 14751380 TI - Initial report of retroperitoneoscopic renal surgery in a patient with factor VIII deficiency (classic hemophilia). AB - Bleeding disorders are generally considered absolute contraindications to laparoscopic surgery. We believe laparoscopic/retroperitoneoscopic renal surgery is an optimal choice in patients with hemophilia, because it minimizes tissue trauma. We present a case of successful retroperitoneoscopic renal surgery in a patient with factor VIII deficiency with a perinephric abscess that failed less invasive treatments. A fully equipped coagulation laboratory, experienced hematologists, and an experienced laparoscopic surgeon play a large role in the outcome in patients with bleeding disorders undergoing minimally invasive procedures. We conclude that although uncorrected bleeding disorders are absolute contraindications for surgical procedures, those that are corrected and appropriately monitored are not. PMID- 14751381 TI - Retroperitoneoscopic radical nephrectomy in a super-obese patient (body mass index 77 kg/m2). AB - Although obesity was initially considered a relative contraindication for laparoscopy, the retroperitoneal approach has been reported to be safe and effective for such patients during renal and adrenal surgery. We report a case of successful retroperitoneoscopic radical nephrectomy in a super-obese patient (body mass index 77 kg/m2) with a 12-cm renal tumor. The operative time was 3 hours, and the estimated blood loss was 100 mL. The patient was discharged home 36 hours after surgery. No intraoperative or perioperative complications occurred. The pathologic examination revealed renal cell carcinoma, and all surgical margins were negative (pT2N0M0). The patient returned to normal activities 3 weeks postoperatively. PMID- 14751382 TI - Bilateral guaifenesin ureteral calculi. AB - We report on a patient with bilateral ureteral calculi composed of guaifenesin metabolite as determined by infrared spectroscopy. These stones may be associated with excessive guaifenesin intake related to the current popularity of ephedrine preparations. PMID- 14751383 TI - Dorsal urethral fistula: case report and review of literature. AB - Urethral fistula is a rare, but recognized, entity. We report the case of a young diabetic patient who developed urethral fistula on the dorsum of penis after debridement for necrotizing fasciitis. This cause and location for urethral fistula is extremely rare, and we were unable to find a similar case in published studies. A brief literature search for various causes of urethral fistula was made, and the likely mechanisms for the cause of the fistula were explored. PMID- 14751384 TI - Fournier's gangrene associated with sparganosis in the scrotum. AB - Fournier's gangrene is a necrotizing fasciitis of the scrotum or perineum that may extend by way of the fascial planes to the penis and the anterior abdominal wall up to the clavicles, buttocks, or lower extremities. It is a life threatening progressive disease that requires aggressive antibiotic therapy and early radical debridement. Sparganosis is a parasitic infection that occurs principally in cats and dogs, but human infestations have been reported, albeit rarely. Recently, we experienced a case of Fournier's gangrene associated with sparganosis in the scrotum, which was treated with antibiotics and extensive debridement including removal of a white, flat, shiny sparganum worm. PMID- 14751385 TI - Xanthogranulomatous orchitis. AB - Xanthogranulomatous inflammation is an uncommon process that is usually associated with severe renal infection. We report a case of a 64-year-old man with xanthogranulomatous inflammation in the testis who presented with contralateral epididymoorchitis and a painless testicular mass. Only two prior cases of xanthogranulomatous orchitis have been reported. This case highlights the need for a broad differential diagnosis when approaching testicular masses, particularly in the setting of coincident infection. PMID- 14751386 TI - Testicular cancer metastatic exclusively to the brain and spleen. AB - We report an unusual case of testicular cancer metastasized solely in the brain and spleen without retroperitoneal disease or other widespread metastasis. The patient initially presented with seizures and underwent craniotomy for presumed primary brain tumor, with a delayed diagnosis of testicular cancer as the true primary. This pattern of spread is highly unusual, and, to our knowledge, we report the first case of splenectomy performed for involvement by testicular cancer. At last follow-up, the patient was doing well, with no evidence of disease after 2 years. PMID- 14751387 TI - Free-floating organized fat necrosis: rare presentation of pelvic mass managed with laparoscopic techniques. AB - Free-floating intraperitoneal pelvic masses are exceedingly rare. These abnormalities, typically composed of organized fat necrosis, are most commonly thought to occur after spontaneous torsion and subsequent infarction of an appendix epiploica. Most of these loose bodies are incidentally discovered. Surgical removal is recommended because cross-sectional imaging typically cannot accurately make the diagnosis. Laparoscopy offers a minimally invasive tool to simultaneously inspect the abdominal cavity, assess the pelvic mass diagnosis, and treat the patient. We describe the case of a patient followed up for pudendal neuropathy in which organized fat necrosis was diagnosed and treated by laparoscopy. PMID- 14751388 TI - Granulocyte colony-stimulating factor receptor signals for beta1-integrin expression and adhesion in bladder cancer. AB - OBJECTIVES: To examine the association of granulocyte colony-stimulating factor (G-CSF) and G-CSF receptor (G-CSFR) expression with increased beta1-integrin expression and determine the ability of autocrine G-CSFR signaling to promote bladder cancer cell adhesion by way of beta1-integrin. beta1-integrin is expressed at higher levels in more invasive bladder carcinoma cells and participates in the process of tissue invasion. Cancer cell invasion and metastasis in some ways mimic normal neutrophil behavior. In neutrophils, G-CSF acts through its receptor to enhance adhesion and migration. A significant fraction of bladder carcinoma has been reported to express both G-CSF and G-CSFR. METHODS: We examined bladder carcinoma tissue samples obtained from segmental or radical cystectomy specimens for expression of G-CSF, G-CSFR, and beta1-integrin using reverse transcriptase-polymerase chain reaction, Western blot, and immunohistochemical methods. We determined the G-CSFR-mediated beta1-integrin adhesion using a static adhesion assay and the bladder cancer cell line 5637. RESULTS: Eleven of 14 bladder cancer samples expressed G-CSFR. All 11 G-CSFR positive tumors also expressed G-CSF, and the G-CSF/G-CSFR positive tumors had elevated beta1-integrin protein levels. All but one G-CSFR negative tumor demonstrated low beta1-integrin protein levels. In four G-CSF/G-CSFR positive tumors for which distant urothelium was available for examination, G-CSF, G-CSFR, and beta1-integrin expression was also increased. In the 5637 cell line, we demonstrated G-CSFR-mediated upregulation of beta1-integrin-dependent adhesion to fibronectin and laminin. CONCLUSIONS: G-CSF/G-CSFR expression in some bladder cancers appears to be an early event during malignant transformation that increases beta1-integrin expression and adhesion and thereby may promote tissue invasion. PMID- 14751389 TI - Nitric oxide release from human corpus cavernosum induced by a purified scorpion toxin. AB - OBJECTIVES: To investigate the effects of a purified scorpion toxin (Ts3) on human corpus cavernosum (HCC) in vitro. Scorpion venoms cause a massive release of neurotransmitters that contribute to the clinical symptoms resulting from envenomation. METHODS: HCC strips were mounted in organ baths containing Krebs solution. After equilibration, the tissues were precontracted with phenylephrine (10 micromol/L). The relaxations caused by Ts3 (30 nmol/L) were compared with those induced by electrical field stimulation (1 to 20 Hz) and nitric oxide (NO, 1 to 100 micromol/L). RESULTS: The addition of Ts3 evoked long-lasting relaxations of precontracted HCC strips, and exogenously applied NO and electrical field stimulation caused short-lived responses. The NO synthesis inhibitor N(omega)-nitro-L-arginine methyl ester (L-NAME; 100 micromol/L) reduced by 87% +/- 2% the Ts3-induced relaxations; this inhibition was reversed by pretreating the tissues with L-arginine (1 mmol/L). The relaxant responses mediated by Ts3 were blocked to a similar degree by the soluble guanylyl cyclase inhibitor 1H-[1,2,4] oxadiazolo [4,3,-alquinoxalin-1-one] (10 micromol/L). In contrast, the addition of the phosphodiesterase type 5 inhibitor sildenafil (0.1 micromol/L) significantly enhanced Ts3-evoked relaxations by 78% +/- 4%. The sodium channel blocker tetrodotoxin (1 micromol/L) completely blocked the relaxant responses elicited by both Ts3 and electrical field stimulation, without significantly affecting those elicited by NO. CONCLUSIONS: The results indicate that Ts3 relaxes the HCC through the release of NO from nitrergic nerves. The elucidation of this mechanism is useful for the development of new therapeutic strategies to treat priapism after scorpion envenomation or to modulate sodium channel activity in the case of penile dysfunction. PMID- 14751390 TI - Congenital unilateral ureteropelvic junction obstruction of the rat: a useful animal model for human ureteropelvic junction obstruction? AB - OBJECTIVES: To investigate the expression of endothelin-1 (ET-1) and adrenomedullin (ADM) in the renal pelvis, stenotic ureteropelvic junction, and ureter of 20 male Wistar rats with congenital unilateral ureteropelvic junction obstruction; the normal contralateral kidneys served as controls. The molecular pathophysiology of congenital ureteropelvic junction obstruction is still unclear. The implication of altered peptidergic innervation is under discussion. Our study group has recently been able to demonstrate a significant increase in ET-1 and a significant decrease in ADM in prestenotic and stenotic tissue, but not in the remainder of the ureter, compared with controls. METHODS: Twenty animals were killed, and samples of the renal pelvis, ureteropelvic junction, upper ureter, middle part of the ureter, and lower ureter were immediately snap frozen and stored in liquid nitrogen. Total RNA was extracted, and subsequently 1 microg of RNA was reversely transcribed. mRNA expression of ET-1 and ADM was determined semiquantitatively using on-line polymerase chain reaction. The expression of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was determined to relate the specific mRNA expression to the expression of a housekeeping gene. RESULTS: We found a significant increase in the expression of ET-1 in the obstructed junctions related to GAPDH (P <0.001). The expression of ADM, however, revealed no statistically significant differences. No differences at all could be detected in the tissue samples from the rest of the ureter. CONCLUSIONS: Alterations in the local production of peptidergic neurotransmitters, especially ET-1, may contribute to the molecular pathogenesis of ureteropelvic junction obstruction. Results previously obtained in the stenotic tissue from children were confirmed in the stenotic tissue from the rat model. We hypothesize that the alterations are disease-, but not age-specific. PMID- 14751391 TI - Protective effect of vasoactive intestinal peptide on testicular torsion detorsion injury: association with heparin-containing mast cells. AB - OBJECTIVES: To elucidate the action of vasoactive intestinal peptide (VIP) on detorsion injury and the heterogeneity of mast cells in the testes of rats. METHODS: Prepubertal male Sprague-Dawley rats were used in six groups. Group 1 was the control group (sham operation); group 2 had 2 hours of torsion; group 3, 2 hours of torsion and 1 hour of detorsion after administration of saline; group 4 had 2 hours of torsion and 4 hours of detorsion after administration of saline; group 5, 2 hours of torsion and 1 hour of detorsion after administration of intraperitoneal VIP (25 ng/kg); and group 6, 2 hours of torsion and 4 hours of detorsion after intraperitoneal VIP. The 2 hours of torsion was created by rotating the right testis 720 degrees in a clockwise direction. VIP (25 ng/kg) was injected intraperitoneally 1 minute before the 1 and 4 hours of detorsion. At the end of the experiment, catalase enzyme activity was measured polarographically, and superoxide dismutase, malondialdehyde, and protein were measured spectrophotometrically. Nitric oxide was measured by capillary electrophoresis in the testicular tissue. Routine histologic examination of testicular mast cells was done under light microscopy; the histochemistry was also analyzed. RESULTS: Torsion significantly induced oxidative stress, mast cell degranulation, and tissue damage. Detorsion attenuated oxidative stress without any diminution of the histologic damage to the tissue. VIP significantly protected the testicular tissue from detorsion injury. It also inhibited mast cell activity while increasing the heparin content. CONCLUSIONS: VIP can protect testicular tissue from detorsion injury. Heparin-containing mast cells seem to be important mediator cells for this protection. PMID- 14751392 TI - Immunohistochemical comparison of vascular and sinusoidal adherens junctions in cavernosal endothelium. AB - OBJECTIVES: To characterize endothelial cell-to-cell junctions in the sinusoids and microvasculature of the corpus cavernosum. METHODS: Corporal tissue was obtained from 6 potent human subjects, cut into 5-microm cryosections, and double labeled with consecutive applications of primary and secondary antibodies. Laser scanning confocal microscopy identified subcellular localization of endothelial anchoring and adhesion molecules. Fluorescence intensity was rated as strong, weak, or absent by two observers. RESULTS: The cavernosal endothelial adherens junction was composed of vascular endothelial cadherin, alpha-catenin, plakoglobin, vinculin, and the regulatory proteins beta-catenin and ZO-1. Adherens junctions in sinusoids were elongated, redundant, and narrow versus short, dense, linear cell-to-cell contacts in small arterioles and venules. Vinculin expression along the basal interface of the endothelium and stroma was weak in the sinusoids and strong in the arterioles. Definite sinusoidal expression of CD31 and CD34 was noted. P-selectin was only expressed within the cavernosal microvessels. CONCLUSIONS: The regulatory and structural proteins extending from vascular endothelial cadherin provide immunohistochemical evidence of a role for adherens junctions in cavernosal endothelial barrier function and cellular homeostasis. The sinusoidal endothelium has a unique junctional phenotype consistent with its blood trapping function. Differential expression of functional proteins in sinusoidal and microvascular endothelium may reflect segmental variation in hemodynamic exposure to pressure, stretch, or flow. PMID- 14751393 TI - Whitmoreisms: memorable quotes from Willet F. Whitmore, Jr, M.D. PMID- 14751395 TI - Treatment of urinary and fecal incontinence with functional magnetic stimulation. PMID- 14751396 TI - Retroperitoneoscopic nephrectomy ("gravity technique"). PMID- 14751398 TI - Acupuncture ameliorates symptoms in men with chronic prostatitis/chronic pelvic pain syndrome. PMID- 14751401 TI - Stereoselective binding of levosimendan to cardiac troponin C causes Ca2+ sensitization. AB - The effects of the Ca(2+) sensitizer levosimendan and that of its stereoisomer dextrosimendan on the cardiac contractile apparatus were studied using skinned fibers obtained from guinea pig hearts. Levosimendan was found to be more effective than dextrosimendan in this model. The respective concentrations of levosimendan and dextrosimendan at EC(50) were 0.3 and 3 microM. In order to explain the difference in efficacy as Ca(2+) sensitizers, the binding of the two stereoisomers on cardiac troponin C was studied by nuclear magnetic resonance in the absence and presence of two peptides of cardiac troponin I. The two stereoisomers interacted with both domains of cardiac troponin C in the absence of cardiac troponin I. In the presence of cardiac troponin I-(32-79) and cardiac troponin I-(128-180), the binding of both levosimendan and dextrosimendan to the C-terminal domain of cardiac troponin C was blocked and only the binding to the N terminal domain was observable. Differences in the overall binding behavior of the two isomers to cardiac troponin C were highlighted in order to discuss their structure to activity relation. Our data are consistent with the notion that the action of levosimendan as a Ca(2+) sensitizer and positive inotrope relates to its stereoselective binding to Ca(2+)-saturated cardiac troponin C. PMID- 14751403 TI - Differential involvement of mu 1-opioid receptors in dermorphin tetrapeptide analogues-induced antinociception. AB - The involvement of putative mu(1)-opioid receptors in the antinociception induced by the dermorphin tetrapeptide analogues Try-D-Arg-Phe-beta-Ala (TAPA) and Tyr-D Arg-Phe-beta-Ala-NH(2) (TAPA-NH(2)) was determined in mice, using a tail-pressure test and a formalin test. TAPA and TAPA-NH(2) injected i.c.v. and i.t. produced dose-dependent antinociception in both assays. In the tail-pressure test, the antinociception induced by i.c.v. or i.t. injected TAPA, but not TAPA-NH(2), was significantly attenuated by pretreatment with naloxonazine, a selective antagonist for putative mu(1)-opioid receptors. Moreover, naloxonazine also significantly attenuated the antinociception induced by i.c.v. injected TAPA, but not TAPA-NH(2), in the formalin test. In contrast, the antinociception induced by both TAPA and TAPA-NH(2) given i.t. was significantly attenuated by pretreatment with naloxonazine in the formalin test. The present results suggest that TAPA and TAPA-NH(2) should be considered selective agonists for putative mu(1)- and mu(2) opioid receptors, respectively. The C-terminal amidation of TAPA-NH(2) may be critical for distinguishing between putative mu(1)- and mu(2)-opioid receptors. PMID- 14751402 TI - Pharmacology of selective acetylcholinesterase inhibitors: implications for use in Alzheimer's disease. AB - Cholinesterase inhibitors vary in their selectivity for acetylcholinesterase versus butyrylcholinesterase. We examined several cholinesterase inhibitors and assessed the relative role of acetylcholinesterase versus butyrylcholinesterase inhibition in central and peripheral responses to these medications. Donepezil and icopezil are highly selective for acetylcholinesterase, whereas tacrine and heptylphysostigmine demonstrated greater potency for butyrylcholinesterase over acetylcholinesterase. All four compounds increased acetylcholine levels in mouse brains. Dose-response curves for tremor (central effect) and salivation (peripheral effect) showed that donepezil and icopezil possess a more favourable therapeutic index than the nonselective inhibitors, tacrine and heptylphysostigmine. Co-administration of the selective butyrylcholinesterase inhibitor tetraisopropylpyrophosphoramide (iso-OMPA) potentiated peripheral, but not central, effects of the selective acetylcholinesterase inhibitor icopezil. The improved therapeutic index observed in mice with icopezil is due to a high degree of selectivity for acetylcholinesterase versus butyrylcholinesterase, suggesting that high selectivity for acetylcholinesterase may contribute to the clinically favourable tolerability profile of agents such as donepezil in Alzheimer's disease patients. PMID- 14751404 TI - Chronic fluoxetine-induced desensitization of 5-HT1A and 5-HT1B autoreceptors: regional differences and effects of WAY-100635. AB - Desensitization of 5-HT(1A) and 5-HT(1B) autoreceptors is thought to be the mechanism underlying the therapeutic effects of fluoxetine and other selective serotonin reuptake inhibitors when these are administered chronically. The blockade of 5-HT(1A) autoreceptors occurring on administration of a selective serotonin reuptake inhibitor together with a 5-HT(1A) autoreceptor antagonist is responsible for the acute increase in 5-hydroxytryptamine (serotonin, 5-HT) levels observed under these circumstances. The effects of repeated administration of selective serotonin reuptake inhibitors together with 5-HT(1A) receptor antagonists have not been widely studied. In this work, we found that the effects of fluoxetine (5 mg/kg, i.p., daily for 12 days) to desensitize 5-HT(1B) autoreceptors in the frontal cortex, as measured by the effect of the locally administered 5-HT(1B) receptor agonist, 3-(1,2,5,6-tetrahydropyrid-4 yl)pyrrolo[3,2-b]pyrid-5-one (CP 93129), and to desensitize 5-HT(1A) autoreceptors as measured by the action of the 5-HT(1A) receptor agonist, 8 hydroxy-2-(di-n-propylamino) tetralin (8-OH-DPAT; 50 microg/kg, s.c.) to reduce 5 HT levels in cortex, were prevented by concomitant administration of the 5-HT(1A) receptor antagonist, N-[2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl]-N-(2 pyridinyl) cyclohexanecarboxamide (WAY-100635; 0.3 mg/kg, s.c.). 5-HT(1B) receptor activity in the hypothalamus, as measured by the effects of locally administered CP 93129, and 5-HT(1A) autoreceptor activity, as determined by the effects of subcutaneous 8-OH-DPAT to reduce 5-HT levels in hypothalamus, were not altered either by fluoxetine alone or by fluoxetine in the presence of WAY 100635. The data suggest that the regulation of extracellular levels of 5-HT in the cortex and hypothalamus is subject to different autoregulatory mechanisms. PMID- 14751405 TI - Chronic antidepressants reverse cerebrocortical allopregnanolone decline in the olfactory-bulbectomized rat. AB - Olfactory bulbectomy is one of the most validated models of depression. We demonstrate that bilateral removal of the olfactory bulbs in rats produced a significant decline of allopregnanolone content in a select cerebrocortical area which was reversed by chronic (3-week) treatment with three different classes of antidepressant (desipramine, fluoxetine, and sertraline, and venlafaxine). The effects of the chronic antidepressant treatments on allopregnanolone cortical content are observed at a time which typically coincides with the drug's abilities to reverse the behavioral deficits of the bulbectomy syndrome. We therefore propose that normalization of allopregnanolone cerebrocortical levels may contribute to the antidepressant-like profile of these drugs in the olfactory bulbectomized rat model of depression. PMID- 14751406 TI - RC-3095, a bombesin/gastrin-releasing peptide receptor antagonist, impairs aversive but not recognition memory in rats. AB - Bombesin and its mammalian equivalent, gastrin-releasing peptide (GRP), stimulate cell proliferation and are involved in the pathogenesis of several types of human cancer. Bombesin-like peptides also display neuroendocrine activities and regulate neural function. In the present study, we evaluated the effects of the bombesin/GRP receptor antagonist (D-Tpi(6), Leu(13) psi[CH(2)NH]-Leu(14)) bombesin-(6-14) (RC-3095), experimental antitumor drug, on memory in rats. Adult female Wistar rats were treated with an intraperitoneal injection of RC-3095 (0.2, 1.0 or 5.0 mg/kg) 30 min before training in either inhibitory avoidance or novel object recognition tasks. Retention test trials were carried out 1.5 (short term memory) or 24 h (long-term memory) after training. RC-3095 at the doses of 0.2 or 1.0 mg/kg, but not at the dose of 5.0 mg/kg, impaired both short- and long term inhibitory avoidance retention, but did not affect recognition memory. The memory-impairing effect of RC-3095 could not be attributed to alterations in sensorimotor functions. The results show that the antitumor drug/GRP antagonist RC-3095 impairs formation of aversive memory. PMID- 14751407 TI - Different role of endothelin ETA and ETB receptors and endothelial modulators in diabetes-induced hyperreactivity of the rabbit carotid artery to endothelin-1. AB - The influence of diabetes on regulatory mechanisms and specific receptors implicated in the contractile response of isolated rabbit carotid arteries to endothelin-1 was examined. Endothelin-1 induced a concentration-dependent contraction that was greater in arteries from diabetic rabbits than in arteries from control rabbits. Endothelium removal or N(G)-nitro-L-arginine enhanced contractions in response to endothelin-1 only in control arteries, without modifying the endothelin-1 response in diabetic arteries. Indomethacin, furegrelate (thromboxane A(2) inhibitor), or cyclo-(D-Asp-Pro-D-Val-Leu-D-Trp) (BQ-123; endothelin ET(A) receptor antagonist) inhibited the contractions in response to endothelin-1, the inhibition being greater in diabetic arteries than in control arteries. 2,6-Dimethylpiperidinecarbonyl-gamma-methyl-Leu-N(in) (methoxycarbonyl)-D-Trp-D-Nle (BQ-788; endothelin ET(B) receptor antagonist) enhanced the contraction elicited by endothelin-1 in control arteries and displaced to the right the contractile curve for endothelin-1 in diabetic arteries. In summary, diabetes induces hyperreactivity of the rabbit carotid artery to endothelin-1 by a mechanism that at least includes: (1) enhanced activity of muscular endothelin ET(A) receptors; (2) impairment of endothelin ET(B) receptor-mediated nitric oxide (NO) release; and (3) enhancement of the production of thromboxane A(2). PMID- 14751408 TI - Potentiation of anandamide effects in mesenteric beds isolated from bile duct ligated rats: role of nitric oxide. AB - Changes in vascular responsiveness are proposed as the basis for some of the cardiovascular complications in cholestasis. Cholestasis is also associated with accumulation of endogenous opioid peptides and evidence of nitric oxide (NO) overproduction. On the other hand, it is well known that anandamide, an endogenous cannabinoid ligand, causes hypotension and a decrease in systemic vascular resistance. In the present study, the possible role of the cannabinoid system in cholestasis-induced mesenteric vascular bed responsiveness was investigated. Mesenteric arteries of bile duct-ligated and sham-operated rats receiving daily administrations of saline were used for evaluating phenylephrine or anandamide dose-response, acute effects of N(G)-nitro-L-arginine methyl ester (L-NAME, 100 microM), a non-selective inhibitor of NO synthase (NOS), or naltrexone, an opioid receptors antagonist (1 microM). The other groups of bile duct-ligated and sham-operated rats received daily intraperitoneal administration of L-NAME (20 mg/kg/day), aminoguanidine, a selective inducible NOS (iNOS) inhibitor (150 mg/kg/day) or naltrexone (10 mg/kg/day). After 7 days, the superior mesenteric artery was cannulated and the mesenteric vascular bed was perfused according to the McGregor method. Anandamide-induced relaxation was significantly potentiated in mesenteric vascular beds of bile duct-ligated rats. Chronic treatment of bile duct-ligated animals with L-NAME and aminoguanidine blocked this hyperresponsiveness while the hyperresponsiveness was potentiated at large doses of anandamide on chronic treatment of these animals with naltrexone. Although acute L-NAME treatment of mesenteric beds completely blocked the anandamide-induced vasorelaxation in sham-operated rats, this vasorelaxation still was present in bile duct-ligated animals. Anandamide-induced vasorelaxation remained unaffected after acute naltrexone treatment of mesenteric beds in both bile duct-ligated and sham-operated rats. Our results indicate that (1) there is enhanced anandamide-induced vasorelaxation in cholestatic rats, probably due to a defect in cannabinoid or vanilloid receptors and (2) NO overproduction may be involved in cholestasis-induced vascular hyperresponsiveness. PMID- 14751409 TI - Morphine potentiates dextromethorphan-induced vasodilation in rat superior mesenteric artery. AB - The combined action of morphine and dextromethorphan on the superior mesenteric artery was investigated in this study. The artery was cut into rings, placed in a muscle bath and mounted to a force transducer for recording tension. Rings preconstricted with 1 microM phenylephrine produced a dose-dependent relaxation to graded doses of dextromethorphan but showed no response to morphine. An equimolar combination of morphine and dextromethorphan exhibited a marked synergism quantitated by a factor of 3.7 (1.8-7.7, 95% CI). Naloxone, which had no effect on the dextromethorphan dose-response relation, abolished the synergism. Removal of the endothelium produced a slight attenuation of the morphine-dextromethorphan synergism, but the magnitude of this attenuation was the same when dextromethorphan alone was examined in the denuded preparation. In contrast to the marked synergism seen in the mesenteric artery preparation, similar experiments on the carotid artery and the aorta produced only additive interactions. PMID- 14751410 TI - The effects of levosimendan and OR-1896 on isolated hearts, myocyte-sized preparations and phosphodiesterase enzymes of the guinea pig. AB - The concentration dependences of the Ca(2+)-sensitizing and the phosphodiesterase inhibitory effects of levosimendan (the (-) enantiomer of [[4-(1,4,5,6-tetrahydro 4-methyl-6-oxo-3-pyridazinyl)phenyl]hydrazono]propanedinitrile) and its active metabolite, OR-1896 (the (-) enantiomer of N-[4-(1,4,5,6-tetrahydro-4-methyl-6 oxo-3-pyridazinyl)phenyl] acetamide), were compared with their positive inotropic effects to reveal their mechanisms of action in guinea pig hearts. In Langendorff perfused hearts, left ventricular +dP/dt(max) increased by 26+/-4% and 25+/-3% (mean+/-S.E.M.), with EC(50) values of 15+/-2 and 25+/-1 nM for levosimendan and OR-1896, respectively. In permeabilized myocyte-sized preparations, levosimendan and OR-1896 both increased isometric force production via Ca(2+) sensitization (at pCa 6.2), by 51+/-7% and 52+/-6%, with EC(50) values of 8+/-1 and 36+/-7 nM (P<0.05), respectively. Thus, the two molecules could be defined as Ca(2+) sensitizers and positive inotropes with very similar concentration dependences. However, major differences appeared when the phosphodiesterase-inhibitory effects of levosimendan and OR-1896 were probed on the two phosphodiesterase isoforms (phosphodiesterases III and IV) dominant in the left ventricular cardiac tissue. Levosimendan was a 40-fold more potent and a 3-fold more selective phosphodiesterase III inhibitor (IC(50) for phosphodiesterase III=2.5 nM, and IC(50) for phosphodiesterase IV=25 microM, selectivity factor approximately 10000) than OR-1896 (IC(50) for phosphodiesterase III=94 nM, and IC(50) for phosphodiesterase IV=286 microM, selectivity factor approximately 3000). Hence, our data support the hypothesis that levosimendan and OR-1896 both exert positive inotropy via a Ca(2+)-sensitizing mechanism and not via simultaneous inhibition of the phosphodiesterases III and IV isozymes in the myocardium at their maximal free plasma concentrations. PMID- 14751411 TI - Central anti-hypertensive effect of tachykinin NK3 receptor antagonists in rat. AB - Tachykinins are involved in the central autonomic control of blood pressure. In the present study, we examined the i.c.v. cardiovascular effects of several tachykinin receptor antagonists in awake spontaneously hypertensive rats (SHR, 15 weeks old). Results showed that two tachykinin NK(3) receptor antagonists (R-820: 3-indolylcarbonyl-Hyp-Phg-N(Me)-Bzl and SB 222200: (S)-(-)-N-(alpha-ethylbenzyl) 3-methyl-2-phenylquinoline-4-carboxamide) caused a sustained and dose-dependent reduction of blood pressure when injected i.c.v. but not i.v. The stereoselective anti-hypertensive effect of SB 222200 peaked at 3 h and faded at 6 h post injection (if injected at 07:00 h) or had a slower onset and peaked at 8 h post injection (if injected at 13:00 h). The effect of R-820 was maximal at 24 h and lasted up to 48 h post-injection. Both antagonists failed to alter blood pressure in normotensive Wistar-Kyoto rats (WKY) and heart rate was not affected in both strains. The anti-hypertensive effect of SB 222200 was not associated with changes in plasma levels of catecholamines and vasopressin and it remained unchanged in SHR subjected to acute bilateral nephrectomy. In contrast, blood pressure was not affected by tachykinin NK(1) (RP 67580: (+/-) 7,7-diphenyl-2[1 imino-2(2-methoxy-phenyl)-ethyl]perhydroisoindol-4-one(3aR,7aR)) and NK(2) (SR 48968: (S)-N-methyl-N[4-(4-acetylamino-4-phenylpiperidino)-2-(3,4 dichlorophenyl)butyl]benzamide) receptor antagonists. Data suggest that brain tachykinin NK(3) receptors are implicated in the maintenance of hypertension in SHR. Hence, these receptors may represent promising therapeutic target in the treatment of arterial hypertension. PMID- 14751412 TI - Serotonin blockade protects against early microvascular constriction following atherosclerotic plaque rupture. AB - Early microvascular constriction following atherosclerotic plaque rupture may be mediated via serotonin and/or endothelin-1. Atherosclerotic lesions in the rabbit hindlimb underwent plaque rupture, resulting in a rapid reduction of distal flow (7.1+/-0.7 ml/min pre-rupture versus 3.6+/-0.6 ml/min post-rupture, P<0.001) and a rise in distal microvascular resistance (10.5+/-0.9 mm Hg min/ml pre-rupture versus 23.5+/-3.5 mm Hg min/ml post-rupture, P=0.01). Distal microvascular resistance remained elevated following endothelin-1 receptor antagonism and control vehicle, but normalised after serotonin receptor antagonism with ritanserin (10.5+/-0.9 mm Hg min/ml pre-rupture versus 22.2+/-6.0 mm Hg min/ml post-endothelin-1 receptor antagonism [P<0.05] versus 21.6+/-6.2 mm Hg min/ml post-control vehicle [P<0.05] versus 11.6+/-2.0 mm Hg min/ml post-ritanserin [P=NS]). Early antagonism of serotonin receptors protects against distal microvascular constriction following atherosclerotic plaque rupture. PMID- 14751413 TI - Abnormal response to ryanodine in oesophageal striated muscle of spontaneously hypertensive rats. AB - The effects of ryanodine on twitch contraction and basal tension of oesophageal striated muscle were compared between preparations from stroke-prone spontaneously hypertensive rats (SHRSP) and normotensive Wistar Kyoto rats (WKY). Ryanodine (3 x 10(-7) M) augmented the twitch contraction in WKY preparations, butt attenuated it in SHRSP preparations. Rates of contraction and relaxation of twitch contraction, normalized to developed tension, were slightly decreased by ryanodine in both preparations. The effect of ryanodine was not different between WKY and SHRSP preparations. Ryanodine elevated the basal tension in WKY preparations but not in SHRSP preparations. Ryanodine elevated the intracellular Ca(2+) level in both preparations, but the response was significantly less in SHRSP preparations. Resting and action potentials were not significantly different between WKY and SHRSP preparations, while the duration of the action potential was significantly longer in SHRSP preparations. Ryanodine did not alter the resting and action potentials of either preparation. These results suggest that the Ca(2+) handling properties, including the ryanodine receptor, of the sarcoplasmic reticulum are genetically altered in oesophageal striated muscle of SHRSP. PMID- 14751414 TI - Cholinergic regulation of Na+-K+-ATPase activity in rat parotid gland: changes after castration. AB - In this study, we investigated the different signalling pathways involved in muscarinic acetylcholine M(3) receptor-dependent modulation of Na(+)-K(+)-ATPase in parotid glands from normal and castrated rats. Carbachol inhibited the enzyme activity in parotid glands from control rats while it stimulated the enzyme activity in castrated rats. The inhibition of Ca(2+) calmodulin by trifluoperazine abolished the inhibitory effect of carbachol in control rats, while the inhibition of protein kinase C by staurosporine stimulated Na(+)-K(+) ATPase. In castrated rats, trifluoperazine inhibited the carbachol-stimulant effect while staurosporine had no effect. Results indicate that in control glands the activation of a phospholipid-Ca(2+) calmodulin-dependent protein kinase C is responsible for the inhibitory effect of carbachol on Na(+)-K(+)-ATPase activity. In castrated rats, the activation of the enzyme by carbachol is regulated by its Ca(2+) calmodulin-stimulating action, and not by activation of protein kinase C. The activation of the Na(+)-K(+)-ATPase observed in castrated rats resulted in a decrease in carbachol-induced net K(+) efflux and thereby could decrease salivary fluid production. PMID- 14751415 TI - Inhibition of cyclooxygenase-2 reduces the protective effect of hepatocyte growth factor in experimental pancreatitis. AB - Hepatocyte growth factor (HGF) overexpression is observed in experimental and clinical acute pancreatitis. Moreover, previous studies have shown that administration of HGF reduces pancreatic damage in experimental pancreatitis. The aim of our studies was to determine the role of cyclooxygenase-1 and cyclooxygenase-2 in the protective effect of HGF administration against caerulein induced pancreatitis. Acute pancreatitis was induced in rats by infusion of caerulein. HGF was administered twice at the dose 10 microg/kg s.c. The activity of cyclooxygenase-1 and cyclooxygenase-2 was inhibited by resveratrol and rofecoxib, respectively (10 mg/kg). Immediately after cessation of caerulein or saline infusion, pancreatic blood flow, pancreatic cell proliferation, pancreatic prostaglandin E(2) generation, plasma lipase activity, plasma interleukin-1 beta and interleukin-10 concentration were measured and morphological signs of pancreatitis were examined. Expression of cyclooxygenase-1 and cyclooxygenase-2 mRNA transcripts was determined by reverse transcriptase-polymerase chain reaction (RT-PCR). Cyclooxygenase protein production was analyzed by Western blot. Administration of HGF or caerulein alone, or their combination, was without effect on cyclooxygenase-1 mRNA expression in pancreatic tissue. Expression of cyclooxygenase-2 mRNA was increased by HGF and caerulein. The maximal increase in cyclooxygenase-2 mRNA expression was observed when HGF administration was combined with caerulein infusion. A similar effect was observed when we studied the influence of HGF and caerulein on pancreatic cyclooxygenase-2 production, as determined by Western blot. Administration of HGF without induction of acute pancreatitis increased pancreatic prostaglandin E(2) generation and plasma interleukin-10, and this effect was abolished by the cyclooxygenase-2 inhibitor, rofecoxib. Treatment with HGF, during the development of pancreatitis, increased the plasma interleukin-10 concentration and attenuated pancreatic damage, as evidenced by: (a) histological improvement of pancreatic integrity; (b) the partial reversal of the decrease in DNA synthesis and pancreatic blood flow; (c) the reduction in pancreatitis-evoked increase in plasma lipase and interleukin-1 beta. Administration of resveratrol and rofecoxib alone was without effect on the development of pancreatitis. Combination of rofecoxib with HGF reduced the HGF evoked increase in plasma interleukin-10 concentration and pancreatic prostaglandin E(2) generation, and abolished the protective effect of HGF against pancreatic damage in pancreatitis. Resveratrol did not affect the protective effect of HGF. We conclude that: (1) HGF induces cyclooxygenase-2 but not cyclooxygenase-1 expression; (2) inhibition of cyclooxygenase-2 in HGF-treated rats decreases the release of anti-inflammatory interleukin-10, increases the production of pro-inflammatory interleukin-1 beta and reduces pancreatic blood flow; (3) cyclooxygenase-2 activity is necessary for the protective effect of HGF in acute pancreatitis. PMID- 14751416 TI - Perspectives for the development of animal models of bipolar disorder. AB - Bipolar disorder (BD) has been a particularly challenging illness for the development of adequate animal models for neurobiological studies. These difficulties are largely related to the peculiar clinical characteristics of this illness, with an intriguing alternation of mania, depression, euthymia, and mixed states. The etiology and brain mechanisms involved in this several mental illness remain unknown. Preclinical studies with animal models of mania or depression have been developed to evaluate the potential efficacy of new psychotropic drugs and generate information concerning the biochemical effects of these drugs on specific targets. These models try to mimic the behavioral components of mania and depression in human subjects and examine the pharmacological responses and mechanisms of action of potentially new therapeutic agents. The main limitation is that there is currently no model that would mimic mood cyclicity, which is a hallmark feature of BD. Thus, these models do not represent valid paradigms for the study of this illness, because they do not address key questions regarding cyclicity. In this review, we propose that new genetics approaches involving potential animal models of BD are a promising new area for further development. PMID- 14751417 TI - Morphine potentiates the impairing effects of neuroleptics on two-way active conditioned avoidance response in male mice. AB - The dopaminergic and opioid systems have effects on the conditioned avoidance response (CAR), although the possible interaction between these systems on this behaviour has not been studied. The effects of morphine (12.6 mg/kg), haloperidol (0.075 mg/kg), sulpiride (20 mg/kg) and risperidone (0.1 mg/kg) alone as well as morphine combined with these dopamine (DA) antagonists on the acquisition and performance of the CAR were explored in mice. Morphine increased avoidances but this seemed secondary to a rise in activity levels. All DA antagonists impaired CAR in the acquisition phase but only haloperidol disrupted performance. The combination of morphine plus neuroleptics impaired acquisition and performance of CAR. These results suggest that morphine disrupts the learning of CAR and that the classical neuroleptic haloperidol profoundly impairs acquisition and performance of CAR to a greater degree than atypical neuroleptics such as sulpiride and risperidone. Finally, it is concluded that morphine potentiates the impairing effects of DA antagonists on CAR. PMID- 14751418 TI - Microinjection of the nitric oxide synthase inhibitor L-NAME into the lateral basal forebrain alters the sleep/wake cycle of the rat. AB - The effects of N(G)-nitro-L-arginine methyl ester (L-NAME) (0.19-0.74 micromol), a competitive inhibitor of the enzyme nitric oxide synthase (NOS); L-arginine (48.0-191.0 nmol), a nitric oxide (NO) precursor; and molsidomine (0.06-0.24 nmol), an NO donor, on spontaneous sleep were studied in adult rats implanted for chronic sleep recordings. Direct bilateral application of L-NAME into the nucleus of the horizontal limb of the diagonal band of Broca (HDB) increased waking (W) and reduced slow wave sleep (SWS). On the other hand, intra-HDB injection of L arginine or molsidomine induced slight but inconsistent changes of sleep variables that did not attain significance. Pretreatment with L-arginine (191.0 nmol) or molsidomine (0.24 nmol) prevented the increase of W and the reduction of SWS induced by L-NAME (0.37 micromol), thus indicating that a decrease in the availability of NO may be involved in the effects of L-NAME on sleep variables. An increase in the release of acetylcholine (ACh) and/or a reduction in the output of gamma-aminobutyric acid (GABA) and adenosine could tentatively explain the effects of L-NAME on SWS and W. PMID- 14751419 TI - Anxiogenic-like activity of 3,4-methylenedioxy-methamphetamine ("Ecstasy") in the social interaction test is accompanied by an increase of c-fos expression in mice amygdala. AB - 3,4-Methylenedioxymethamphetamine (MDMA) is a synthetic amphetamine popularly known as "Ecstasy." Animal studies examining acute effects of MDMA on anxiety are unclear because although an anxiolytic-like action of MDMA in different animal models of anxiety has been described, there is also substantial evidence supporting an anxiogenic-like effect of this drug. To date, several studies have examined c-fos expression following MDMA administration in rats. However, there is no information about the MDMA-induced c-fos expression in mice previously tested in an animal model of anxiety. In this study, male mice were injected with MDMA (1, 8 and 15 mg/kg ip) and assessed for changes on anxiety and for the expression of the immediate early gene c-fos in the amygdala (central, basolateral and basomedial). Anxiety was evaluated by the "social interaction test." Ten behavioral categories were recorded: body care, digging, nonsocial exploration, exploration from a distance, social investigation, threat, attack, avoidance/flee, defense/submission and immobility. As compared with the control group, mice treated with MDMA (all doses) showed a decrease in mean duration and total time spent in social investigation behaviors, whereas avoidance/flee behaviors were significantly increased after treatment with this compound (8 and 15 mg/kg). Likewise, a significant increase in c-fos expression was found in the basolateral (all doses) and central (15 mg/kg) amygdala after MDMA administration. Overall, these findings indicate that MDMA exhibits an anxiogenic like profile in the social interaction test in mice, and that central and basolateral amygdala might be involved in these anxiogenic-like effects of the drug. PMID- 14751420 TI - Is vitamin D hypothesis for schizophrenia valid? Independent segregation of psychosis in a family with vitamin-D-dependent rickets type IIA. AB - The vitamin D hypothesis of schizophrenia is a recent concept bringing together old observations on environmental risk factors and new findings on the neurodevelopmental effects of vitamin D. Candidate genes related to the vitamin D endocrine system have not yet been fully explored for this purpose. The coexistence of vitamin-D-dependent-rickets type II with alopecia (VDDR IIA) and different forms of psychosis in the same inbred family has provided us with an opportunity to investigate the presumed relationship between vitamin D deficiency and psychosis. Psychiatric examination and molecular genetic studies were performed in this family overloaded with psychotic disorders and VDDR IIA. Forty members were evaluated in order to describe their phenotypic features. The family was tested for a linkage to the chromosome 12q12-q14 region where the vitamin D receptor (VDR) gene is located. Psychosis was the common phenotype in the 18 psychiatrically affected members. Pedigree analysis did not show a cosegregation of psychosis and rickets. Lod scores were not significant to prove a linkage between psychosis and VDR locus. The authors concluded that (1) the neurodevelopmental consequences of vitamin D deficiency do not play a causative role in psychotic disorders, (2) these two syndromes are inherited independently, and (3) vitamin D deficiency does not act as a risk factor in subjects susceptible to psychosis. PMID- 14751421 TI - Clinical profile of responders to buprenorphine as a substitution treatment in heroin addicts: results of a multicenter study of 73 patients. AB - In France, high-dosage buprenorphine (HDB) is the main substitution treatment for narcotic addiction. Few data have been published concerning clinical factors predicting a good response to this treatment in a daily practice. A hospital based multicenter clinical research program (PHRC) was undertaken in heroin addicted patients, diagnosed according to DSM-III-R, to detect clinical criteria susceptible of predicting a good response to HDB administered during a 3-month treatment period. At the inclusion time in the study, a diagnostic structured interview (DIGS) was performed, and the Addiction Severity Index (ASI), Zuckerman scale, depression scale from Jouvent, and CGI were scored. MMPI was also administered. Good response was defined as an ongoing participation in the study, with absence of opiate detected in 75% of urine collected during the last month of treatment. Only subjects treated for at least 1 month were eligible for analyses. One hundred fifteen patients were recruited and 73 were analyzed. Patients received 8.5+/-2.6 mg (m+/-S.D.) of buprenorphine for 1 to 3 months. A forward stepwise logistic regression showed that six clinical parameters may predict a good response to treatment: probability to respond to buprenorphine was higher in subjects having a high psychopathology (ASI) subscore, low disinhibition and boredom susceptibility factor scores (Zuckerman scale), no alcohol dependence, no family history of addiction or mood disorder, and duration of opiate dependence less than 10 years. Only the MMPI D subscale was a psychological pattern correlated to a good response to substitution treatment. These findings are important to consider when making the decision to prescribe HDB substitution treatment in opiate addiction. PMID- 14751422 TI - Neurons expressing calcium-binding proteins in the prefrontal cortex in schizophrenia. AB - Increased neuronal density, cortical thinning, and alterations of GABAergic interneurons in the prefrontal cortex have been associated with the pathophysiology of schizophrenia. This study used antibodies directed against the calcium-binding proteins, calretinin (CR), parvalbumin (PV), and calbindin (CB) to compare the relative density of subpopulations of GABAergic interneurons in BA9 of the prefrontal cortex from six subjects with schizophrenia and six control subjects matched for age, gender, and postmortem interval. The relative density of interneurons expressing CR, PV, or CB did not differ significantly between subjects with schizophrenia and control subjects. In addition, no change in somal size of immunoreactive (IR) neurons or cortical thickness was observed between the two groups. This study supports previous reports consistently demonstrating no change in the relative density of interneurons expressing CR in the dorsolateral prefrontal cortex in schizophrenia but does not support previous inconsistent findings that the relative density of interneurons expressing PV and CB might be altered in this disorder. PMID- 14751423 TI - Hypothalamic-pituitary-adrenal axis function following a 35% CO2 inhalation in healthy volunteers. AB - RATIONALE: The hypothalamic-pituitary-adrenal axis (HPA axis) is a central component of the brain's neuroendocrine response to stress. The extent of increase in cortisol secretion, provides an index of the HPA axis activity, and in this way, objectively reflects perceived stress. In healthy subjects, the 35% CO(2) inhalation does hardly induce stress, as expressed in anxiety. However, inconsistent results have been found in studies investigating the cortisol response following CO(2) inhalation. Clarity has to be reached about the normal reaction to this challenge, especially because this model is still a very valuable method to study central aspects of panic. OBJECTIVES: The present study aimed to test the hypothesis that a single breath of 35% CO(2) would not induce cortisol release in healthy volunteers. METHODS: In the current study, 20 healthy subjects underwent both a 35% CO(2) and a placebo inhalation in a randomised, single blind fashion. Cortisol levels were determined in saliva samples, taken at regular intervals. RESULTS: No differences were found between the CO(2) and the placebo condition. In both conditions a significant time effect was found, which can be subscribed to normal variation in the circadian rhythm. Furthermore, only modest subjective anxiety scores were found in the CO(2) condition. CONCLUSIONS: These results provide biological evidence for the hypothesis that healthy subjects are not affected by the 35% CO(2) challenge in a clinically significant way. Characteristic, PD patients react much stronger to the inhalation. Thus, in addition to psychological parameters, healthy subjects also constitute an ideal comparison group with regard to endocrinological parameters. PMID- 14751424 TI - Adverse effects of risperidone and haloperidol treatment in schizophrenia. AB - PURPOSE: Side effects of pharmacological treatment in schizophrenia continue to be a major issue in spite of the development of new antipsychotics. The aim of this study is to explore the adverse effects of conventional and atypical antipsychotic drugs and their associated factors. METHODS: Over 3 months, 41 patients with schizophrenia were randomized to treatment with risperidone 1-12 mg (n=21) or haloperidol 2-20 mg (n=20) daily. Efficacy was assessed by improvement of psychotic symptoms, measured on the Positive and Negative Syndrome Scale (PANSS). The safety and tolerability were assessed with the Extrapyramidal Symptom Rating Scale, the UKU Side-Effect Rating Scale and clinical laboratory assessments. RESULTS: Each treatment reduced psychotic symptoms. PANSS total scores, positive scores, and general psychopathology scores declined as trial went on without significant differences between the two groups. While PANSS negative scores improved better in the risperidone group than in the haloperidol group. The tolerability of antipsychotics was statistical significantly better in the risperidone than in the haloperidol-treated patients. The most frequent adverse effects for both groups were tremor and rigidity. Antipsychotics, their doses, and hyperprolactinemia predict short-term extrapyramidal side effects. Serum prolactin levels could predict parkinsonism and dyskinesia severity. However, dyskinesia was best predicted by the doses of neuroleptics. The predictive factor of dystonia was the antipsychotic drug itself. After adjusting drug doses and concomitant medications, side effects could be markedly improved. CONCLUSIONS: This study suggested that risperidone was superior to haloperidol in improving negative symptoms and better tolerated during the 12 weeks' treatment of schizophrenia. Serum prolactin levels could predict the severity of parkinsonism and dyskinesia. PMID- 14751425 TI - Effects of the vanilloid agonist olvanil and antagonist capsazepine on rat behaviors. AB - Vanilloid receptors (VR) are molecular integrators of painful chemical and physical stimuli. Olvanil is an agonist of the vanilloid receptor; capsazepine is a competitive VR antagonist. The authors were interested in investigated the effects of these compounds on anxiety-like behaviors in rats using the elevated plus maze. In addition, the authors examined the effects of olvanil on the Porsolt swim test. Doses of 0, 0.2, 1.0 and 5.0 mg/kg olvanil, respectively, yielded percent open arm entries at 5 min of 25+/-10.1, 19.3+/-7.1, 14.9+/-5.9 and 0+/-0. We demonstrated a drug effect by showing that the mean of the 0.2, 1 and 5 mg/kg doses was significantly lower than the 0 mg/kg dose at P<.05. In addition, the authors examined the effect of olvanil on the ability of rats to perform in the Porsolt swim test. Float time for rats was tested with 0.1 or 2 mg/kg olvanil and differences between the float times for the lower and higher doses were significant at P<.05. In addition, the effects of various doses of the vanilloid antagonist capsazepine was examined on elevated plus maze behavior. Doses of 0, 1, 5 and 10 mg/kg yielded percent time in the open arms at 5 min of 1.46+/-1.38, 15.05+/-10.42, 11.54+/-10.57, and 14.56+/-7.86. The mean of the 1, 5 and 10 mg/kg doses was significantly greater than the percent time in the open arms for the vehicle, consistent with a drug effect. The results suggest that the vanilloid agonists and antagonists may impact on behaviors involving anxiety and affect. However, it cannot be ruled that the findings could be due to nonspecific motor effect. PMID- 14751427 TI - Effects of antidepressant treatment on thyrotropin-releasing hormone stimulation, growth hormone response to L-DOPA, and dexamethasone suppression tests in major depressive patients. AB - Dexamethasone suppression (DST), thyroid-stimulating hormone (TSH) and prolactin (PRL) responses to thyrotropin-releasing hormone (TRH) and growth hormone (GH) response to L-DOPA tests were evaluated in 19 depressed inpatients before the commencement of the antidepressant treatment and after the clinical response to examine: (i) the functional relationships among the hypothalamic-pituitary adrenal (HPA) and hypothalamic-pituitary-thyroid (HPT) axis and dopaminergic system in depression, (ii) any alterations in these hormonal functions with the antidepressant treatment. TSH responses to TRH showed a tendency to increase from pre- to posttreatment period, while TRH-induced PRL and L-DOPA-induced GH responses did not change with treatment in depressed patients who responded to the treatment. Females showed significantly higher TSH and PRL responses to TRH compared to males. No interconnections were found among the responses in DST, TRH stimulation test and L-DOPA-induced GH test in the patients. The results do not support the interrelations between the abnormalities in the HPT and HPA axes and central dopaminergic activity in depression. PMID- 14751426 TI - Effects of clozapine and its metabolites on the 5-HT2 receptor system in cortical and hippocampal cells in vitro. AB - PURPOSE: The goal of the present study was to determine the effects of clozapine (Cloz) and its metabolites norclozapine (Norcloz) and clozapine-N-oxide (Cloz-N oxide) on the 5-HT(2) receptor system on the levels of protein and gene expression in in vitro systems of primary cortical cells of the rat and human hippocampal SHS5Y5 neuroblastoma cells. METHODS: Clinically relevant concentrations of Cloz (200/400 ng/ml) and its metabolites (200 ng/ml) were used for the examination of the effects of Cloz and its metabolites on serotoninergic 5-HT(2) receptor parameters (density, affinity and mRNA levels) as well as on glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA levels in primary cortical cells of the rat after treatment for 24 h under in vitro conditions. To compare the results to human cells, we also measured treatment-induced changes in 5-HT(2) and GAPDH mRNA levels in human hippocampal SHS5Y5 cells. RESULTS: A significant decrease was found in primary cortical cells for 5-HT(2) receptor density (Cloz 200/Cloz 400/Norcloz 200 and Cloz-N-oxide 200 vs. control) and 5-HT(2A) receptor mRNA levels (Cloz 200 vs. control). 5-HT(2A) receptor mRNA levels were also significantly reduced (Norcloz 200 vs. control) in SHS5Y5 cells. GAPDH mRNA levels were not affected. CONCLUSIONS: The results of the present study show that Cloz and Norcloz induce significant alterations on the 5-HT(2) receptor system in primary cortical cells of the rat and in human hippocampal cells. PMID- 14751428 TI - Double-blind, randomized comparison of olanzapine versus fluphenazine in the long term treatment of schizophrenia. AB - This study was undertaken to evaluate the efficacy and safety of olanzapine compared with fluphenazine in the treatment of patients who met the Diagnostic and Statistical Manual, fourth edition (DSM-IV) diagnostic criteria for schizophrenia or schizoaffective disorder. This was a long-term (22-week), randomized, double-blind, parallel clinical trial. Sixty patients (mean age, 35.4 years) were randomly assigned to either olanzapine (n=30) or fluphenazine (n=30). They received treatment at three centers in Croatia during a 22-week study period and were assessed weekly for the first 6 weeks and monthly thereafter. Efficacy was measured using the Brief Psychiatric Rating Scale (BPRS), the Positive and Negative Syndrome Rating Scale (PANSS) and the Clinical Global Impression (CGI) Severity and Improvement scores. The Hillside Akathisia Scale (HAS), Simpson Angus Scale (SAS), Abnormal Involuntary Movement Scale (AIMS), vital signs, laboratory tests, and treatment-emergent adverse events were assessed to evaluate safety. The olanzapine group showed significantly greater mean decreases from baseline to endpoint for BPRS total (-25.8 vs. -16.5, P=.035), PANSS total (-45.7 vs. -29.5, P=.037), PANSS positive (-13.0 vs. -7.9, P=.034), and CGI Severity ( 2.2 vs. -1.3, P=.031) scores. The olanzapine group showed greater mean decreases on all measures of extrapyramidal symptoms, significantly so for the SAS (-2.1 vs. 1.9, P=.004) and HAS (-3.4 vs. 2.6, P=.028). Patients in the fluphenazine group experienced a higher incidence of treatment-emergent adverse events (76.7% vs. 50.0%, P=.032). Weight gain was the most frequently reported adverse event in the olanzapine group (16.7% vs. 0.0%, P=.020). Akathisia (30.0% vs. 10.0%, P=.053) and insomnia (20.0% vs. 0.0%, P=.010) appeared most frequent in the fluphenazine group. Daily use of anticholinergics and benzodiazepines were both significantly greater for the fluphenazine group (P=.003 and.04, respectively). No significant changes were observed in vital signs, ECG, or clinical chemistry. The study indicates that olanzapine has advantages in both efficacy and safety compared to fluphenazine; however, the small sample size limits our ability to draw definitive conclusions. PMID- 14751429 TI - Regional cerebral blood flow and plasma nicotine after smoking tobacco cigarettes. AB - The hypothesis for this research is that only in some brain areas, regional cerebral blood flow (rCBF) after tobacco smoking is correlated with arterial plasma nicotine concentrations. Twenty-one healthy adult tobacco smokers of both genders were studied after overnight tobacco abstinence. H(2)15O water was used to measure rCBF. Six separate scans were taken about 12 min apart with the subjects' eyes closed and relaxed. Research tobacco cigarettes smoked were of average (1.0 mg) and low (0.08 mg) nicotine but similar tar yield (9.5 and 9.1 mg). Increases in normalized rCBF were obtained in the occipital cortex, cerebellum, and thalamus, and decreases in the anterior cingulate, nucleus accumbens, amygdala, and hippocampus immediately after smoking the first average nicotine yield cigarette of the morning. After smoking the second average nicotine yield cigarette, the effects were less than smoking the first. Low nicotine cigarettes produced fewer changes in rCBF than those after the first average cigarette. As expected, statistically significant correlations were found between increases in arterial plasma nicotine and HR. Correlations with arterial nicotine on rCBF were statistically significant in brain areas with the greatest changes in relative blood flow such as the cerebellum and occipital cortex. Nicotine delivery by tobacco smoking is only one of the factors, which contribute to changes in rCBF. PMID- 14751431 TI - Spontaneous firing rate of lateral septal neurons decreases after forced swimming test in Wistar rat. AB - The systemic or local administration of diverse antidepressants increases the neuronal firing rate of the lateral septal nucleus (LSN), whereas some stressful situations decrease its firing rate; however, any long-lasting effect exerted by the forced swimming (FS) test (15-min pretest and 5-min test 24 h later) on the firing rate of the LSN is unknown. Therefore, single-unit extracellular recordings were obtained from the LSN neurons of control rats (Ctrl, n=6) and FS rats (n=10) 2 h after the last swimming session. In other rats, spontaneous firing rate of cortical neurons was recorded under the same experimental conditions. The firing rate of the LSN neurons of the animals in the FS group was significantly lower (9.2+/-1.7 spikes/10 s; P<.004, n=35) in comparison with the Ctrl group (21.1+/-3.4 spikes/10 s, n=22). The reduced firing rate in the LSN after swimming tests was both evident and generalized given that approximately 83% of the total recorded neurons from the FS group fired below the mean+/-1 S.D. rate obtained from the Ctrl group. Accordingly, the mean first-order interval of neuronal firing rate in the FS group (621.3+/-22.6 ms) was significantly greater (P<.05) than that observed in the Ctrl group (391.5+/-29.2 ms), but no significant differences were found in the variation coefficient of these two experimental groups, illustrating regularity of firing. Nonsignificant differences or even an opposite trend were observed in the firing rate of cortical neurons in the FS group (26.3+/-8.4 spikes/10 s) as compared with Ctrl group (15.4+/-1.1 spikes/10 s). Accordingly, no differences were found in the variation coefficient (FS 55.3+/-7.2%, Ctrl 55.8+/-3.6%) or average first-order interval (FS 417.8+/-71.8 ms, Ctrl 494.1+/-64.5 ms). We conclude that the FS test constitutes a situation whose capacity for inducing long-lasting despair is reflected in a reduction in the firing rate of LSN neurons as it occurs in situations of anxiety and fear, contrary to the actions of antidepressant drugs. PMID- 14751430 TI - Neurocognitive function in borderline personality disorder. AB - A battery of neuropsychological measures considered sensitive to dysfunction in prefrontal or temporal cortices was administered to patients with borderline personality disorder (BPD) and healthy controls. BPD patients exhibited striking deficits on measures of nonverbal executive function and nonverbal memory but were unimpaired on tests of alternation learning, response inhibition, divergent thinking, verbal fluency, and verbal working memory. A second study found that university students obtaining high scores on a self-report measure of BPD symptoms exhibited a similar pattern of neuropsychological impairment, although performance deficits were much less pronounced in the student sample. Taken together, these studies suggest that dysfunction of a right hemisphere frontotemporal regions may be associated with borderline personality. PMID- 14751432 TI - Emotional withdrawal, CT abnormalities and drug response in late life depression. AB - In this study, the authors investigated if CNS degenerative abnormalities could correlate with depressive symptoms in elderly patients, if the presence of mild/moderate cognitive impairment could be related to the response to treatment and the role of peculiar clinical features in influencing the response to treatment. Fifty-three patients (60-75 years) diagnosed as affected by late onset (after 60 years) Major Depressive Episodes according to DSM-IV criteria were studied. Brain vascular and degenerative markers were assessed by computed tomography (CT) through measurements of a lateralized version of the bifrontal index and a rating scale addressing subcortical disease. The presence of mild/moderate cognitive impairment [(24-28 total score at the Mini-Mental State Examination (MMSE)], and of specific symptoms were assessed at baseline and evaluated with respect to the antidepressant response. Patients with CT abnormalities showed higher baseline scores on Hamilton Rating Scale for Depression (HAM-D) items "late insomnia" (t=-2.674, P=.002), "somatic symptoms" (t=-3.355 P=.002), and Brief Psychiatric Rating Scale (BPRS) item "emotional withdrawal" (t=-3.355, P=.002). No significant correlation was found between the vascular index and baseline clinical symptoms, while the HAM-D "depressed mood" item was negatively correlated to the right frontal index (R=-0.692, P=.006). Patients with CT abnormalities showed a lower reduction of HAM-D total scores than patients with normal CT (time effect: F=29.277, P<.0001; group effect: F=5.154, P<.03), while a significant reduction of symptoms in time (time effect: F=33.33, P<.0001) but no differences between groups were found on Hamilton Rating Scale for Anxiety (HAM-A). Both patients with and without mild cognitive impairment improved on the HAM-D (time effect: F=19.668, P<.0001), BPRS (time effect: F=18.345, P<.0001), and HAM-A (time effect: F=17.959, P<.0001) total scores. Patients with emotional withdrawal showed lower improvement on BPRS total scores (time effect: F=26.946, P<.0001; group effect: F=5.121, P<.03). The results from this study showed that patients with baseline emotional withdrawal and CT abnormalities have poorer outcome. Further investigations on larger samples are needed to confirm these findings. PMID- 14751433 TI - Selective antagonist [3H]SR141716A binding to cannabinoid CB1 receptors is increased in the anterior cingulate cortex in schizophrenia. AB - Previous studies suggest that long-term cannabis use causes cognitive impairment, including lack of motivation and impaired attention, conditions that also resemble core negative symptoms of schizophrenia. The anterior cingulate cortex (ACC) plays an important role in normal cognition, particularly in relation to motivation and attention. This could suggest that changes in the cannabinoid (CB) system might be present in the ACC of patients suffering schizophrenia. The present study examined the distribution and density of CB1 cannabinoid receptors in the left ACC taken postmortem from patients with schizophrenia (n=10) and matched control subjects (n=9). Radioligand binding of [3H]SR141716A, an antagonist that specifically targets CB1 receptors of the endogenous cannabinoid system, was examined on ACC sections using quantitative autoradiography. CB1 receptors had a homogeneous distribution among the layers of ACC. A significant 64% increase in [3H]SR141716A specific binding to CB1 receptors was found in the schizophrenia group as compared to the control group (mean+/-S.E.M.: 46.15+/-6.22 versus 28.02+/-4.20 fmol/mg estimated tissue equivalents; p=0.03). The present results support the suggestion that changes in the endogenous cannabinoid system in the ACC may be involved in the pathology of schizophrenia particularly in relation to negative symptoms. PMID- 14751434 TI - A clinical case series of switching from antipsychotic polypharmacy to monotherapy with a second-generation agent on patients with chronic schizophrenia. AB - Second-generation antipsychotic medications have become popular as a treatment for schizophrenia. The authors investigated 25 chronic subjects who had previously been treated with high-dose antipsychotic polypharmacy without amelioration. All patients had a history of having been treated with an antipsychotic polypharmacy regimen of the total daily chlorpromazine equivalent dose exceeding 1000 mg/day for more than 6 weeks. They were subsequently switched to a second-generation antipsychotic monotherapy. Other psychotropic medications were simplified at the same time. For successful patients whose symptoms showed at least minimal improvement, the medical chart was reviewed retrospectively. After completed switching, the patients were followed up for 12 weeks, when final evaluation was made. The Global Assessment of Functioning score improved from 32 to 47. The number of antipsychotic medications and total psychotropic medications were significantly reduced from 3.5 to 1.1 and 6.8 to 2.6, respectively. Also, the antipsychotic dose was significantly minimized from 2203 to 619 mg/day. Eleven of eighteen inpatients were discharged and the other four were better enough to be ready for discharge. By showing successful cases, the authors suggest a possibility of antipsychotic monotherapy with a second-generation agent even for those patients who had been treated with high-dose antipsychotic polypharmacy in vain. PMID- 14751435 TI - Heart rate dynamics and their relationship to psychotic symptom severity in clozapine-treated schizophrenic subjects. AB - The analysis of heart rate variability (HRV) has proven to be useful in evaluating the neuroautonomic dysfunctions associated with various clinical conditions. The purpose of this study was to investigate the linear and non linear dynamic measures of HRV, and to evaluate their relationship with the psychotic symptom severity, in clozapine-treated schizophrenic subjects. Fifty schizophrenic patients treated with clozapine as monotherapy and 50 normal control subjects were evaluated for HRV analysis. The HRV measurements were obtained from a 30-min resting electrocardiogram (ECG). The severity of psychotic symptoms was assessed using the Positive and Negative Syndrome Scale (PANSS). In the patient group, the complexity and symbolic dynamics measures as well as the time and frequency domain measures of HRV were significantly lower than in the control group (P<0.01). The intermediate-term fractal scaling component value was significantly higher in the patient group (P<0.01). The PANSS total score and the positive symptom subscale score had significant negative correlations with the sample entropy (SampEn) value (P<0.01). In conclusion, schizophrenic patients treated with clozapine had markedly different heart rate dynamics compared to normal control subjects. The severity of psychotic symptoms was associated with the SampEn value, suggesting that the non-linear complexity measure might be useful in assessing the neuroautonomic dysfunction in schizophrenia. PMID- 14751436 TI - Distribution of neuropeptide Y interneurons in the dorsal prefrontal cortex of schizophrenia. AB - The distribution of neuropeptide Y (NPY) containing neurons was investigated in the dorsal prefrontal region in the brains of the schizophrenic patients and compared to those of normal control. Proportional comparison of NPY neurons in four compartments, upper cortical layers, lower cortical layers, subcortical white matter and deep white matter, demonstrated differential distribution between schizophrenic brains and controls. The proportion of NPY neurons in the upper cortical layers was low in disorganized form and subsequently in paranoid form in comparison to controls. The proportion of NPY neurons in the deep white matter was, conversely, high in the disorganized form and subsequently in the paranoid form. These results indicate that there may be a gamma-aminobutyric acid (GABA)-ergic deficit in schizophrenic patients, especially, in the disorganized form. These results also support the hypothesis of neurodevelopmental dysfunction of schizophrenia. PMID- 14751437 TI - Possible role of D-serine in the pathophysiology of Alzheimer's disease. AB - Several lines of evidence suggest that D-serine may function as an endogenous agonist of the glycine site on the N-methyl-D-aspartate (NMDA) receptor that has been implicated in the pathophysiology of Alzheimer's disease (AD). The purpose of the study was to determine whether serum levels of D- and L-serine in patients with AD are altered as compared with normal controls. Serum levels of D- and L serine in patients of AD and age- and gender-matched normal controls were determined using a high-performance liquid chromatography (HPLC). Serum levels of D-serine in the patients with AD were slightly (z=-1.77, p=0.078) lower than those of normal controls. In contrast, serum levels of L-serine in the patients were slightly (z=-1.73, p=0.083) higher than those of controls. In addition, the percentage (%) of D-serine in the total (L+D) serine in the patients was significantly (z=-2.36, p=0.018) lower than that of controls. The present study suggests that the reduced activity of serine racemase, an enzyme catalyzing the formation of D-serine from L-serine may play a role in the pathophysiology of AD. PMID- 14751438 TI - Blunted inhibition of return in schizophrenia-evidence from a longitudinal study. AB - Previous cross-sectional studies on covert orienting of visual attention in schizophrenia have been inconsistent. In the present longitudinal study, we examined 40 medicated acutely ill inpatients with a covert orienting of attention task (COVAT) shortly after admission, and again 12-16 weeks after the initial examination, while most patients were in (partial) remission. We administered a COVAT with nonpredictive peripheral cues and two stimulus-onset asynchronies (SOA; 100 and 800 ms). In addition, we examined 34 healthy control subjects twice (2 weeks apart). The most important finding was a lack of inhibition of return (IOR) in patients with schizophrenia, both at the first examination in an acute psychotic state and at the follow-up examination after considerable clinical improvement. The IOR deficit was unrelated to psychopathology, length of illness, number of previous psychotic episodes, and type of neuroleptic (NL) medication. Deficient IOR in patients with schizophrenia appears to be state-independent and might be viewed as a trait or vulnerability marker of the disorder. Subsequent studies with never-medicated populations and with schizotypal or high-risk subjects are needed in order to further analyze the possible role of NL medications and to clarify whether blunted IOR might represent a vulnerability marker of schizophrenia. PMID- 14751439 TI - Neuroanatomy of coprolalia in Tourette syndrome using functional magnetic resonance imaging. AB - OBJECTIVE: To determine the neural substrates of phonic tics in Tourette syndrome (TS) using functional magnetic resonance imaging (fMRI) and compare with a proposed tic-generating network (TGN). PATIENTS: One with TS and one normal control. METHODS: fMRI scans were obtained on the TS patient during which numerous unsuppressed phonic tics occurred and, along with the scanner noise, were recorded on audiotape. The control underwent the same functional MRI sequence but mimicked the tics within predetermined, on-off time blocks. Fuzzy clustering (FC) methods were used to generate the activation maps. RESULTS: The TS patient and control showed fMRI activation in the left middle frontal gyrus and right precentral gyrus. The TS patient also had activity in the caudate nucleus, cingulate gyrus, cuneus, left angular gyrus, left inferior parietal gyrus, and occipital gyri. CONCLUSIONS: fMRI, using an FC analysis, is a viable technique for studying TS patients with phonic tics. These results give further support to the hypothesis of a tic-generating circuit model. Further studies are required to confirm our data. PMID- 14751440 TI - Aging and visual hallucinations in elderly psychiatric outpatients. AB - Estimates of the reported prevalence of visual hallucinations in the elderly vary from study to study. The aim of the present investigation was to re-evaluate the prevalence of visual hallucinations in elderly psychiatric outpatients and to examine pertinent risk factors. One hundred and twenty-two elderly (> or =60 years old) consecutive outpatients at our university hospital were investigated with a questionnaire regarding visual hallucinations and the Mini-Mental State Examination. Twelve of 122 (10%) patients suffered from visual hallucinations. Although advanced age, lower MMSE scores and dementia were significantly associated with the presence of visual hallucinations, logistic regression analysis revealed that only advanced age showed a tendency to predict the presence of visual hallucinations. In 53 patients with dementia examined separately, only advanced age was significantly associated with visual hallucinations. The present findings suggest that the prevalence of visual hallucinations in the elderly is about 10% and that aging is an important risk factor in the development of visual hallucinations in dementia. Further studies are required in order to investigate the contribution of age-related factors such as lowering visual acuity, bereavement and loneliness to the presence of visual hallucinations. PMID- 14751441 TI - Effects of baclofen on feeding behaviour examined in the runway. AB - The motivational mechanisms underlying the effects of systemic administration of the GABA-B agonist baclofen on feeding were examined using a runway. Food deprived male hooded Lister rats were trained to traverse a runway for food reinforcement. Baclofen (1 mg/kg i.p.) significantly increased food intake and this was most evident on the final two blocks of testing. The 2 mg/kg dose of baclofen increased running speed without significantly altering intake. At the highest dose tested (4 mg/kg), no significant effects on either consummatory or appetitive measures were observed. These data suggest that low doses of baclofen enhance the consummatory phase of ingestion by attenuating the natural signals associated with onset of satiation. The data also suggest that baclofen has complex effects on appetitive behaviour that may interfere with its effects on consumption. PMID- 14751442 TI - Lack of association between polymorphic variations in the alpha 3 subunit GABA receptor gene (GABRA3) and suicide attempts. AB - This is the first case-control study exploring the association between suicide attempts and the polymorphic variations of the alpha 3 subunit gamma-aminobutyric acid (GABA) receptor gene (GABRA3) located in chromosome X. In a Spanish general hospital, 184 suicide attempters (127 women and 57 men) and 275 control blood donors (109 women and 166 men) were recruited. The four frequent variants (A1, A2, A3 and A4) of GABRA3 were studied. There were no significant differences in the total or by-gender frequencies of the four alleles. In females, there were no significant differences in the genotypes. This study can rule out even small size effects in the total sample and suggests a lack of association between GABRA3 polymorphism and suicide attempt, in the Spanish population. PMID- 14751443 TI - Weight gain and improvement with quetiapine in bipolar I disorder: a case report. AB - Observations made with quetiapine (QUET) in this case give clues for some aspects of its use for patients with bipolar disorder. Weight gain (11 kg; 16.6% increase in 21 weeks) and improvement in manic symptoms occurred after QUET add-on to lithium (Li). Patient's mood improved after QUET add-on without causing extrapyramidal symptoms (EPS), while QUET was discontinued due to weight gain. Short-term QUET add-on to Li may help mood stabilization in bipolar I disorder. Weight changes must be observed carefully. PMID- 14751444 TI - A case of neurosyphilis showing a marked improvement of clinical symptoms and cerebral blood flow on single photon emission computed tomography with quantitative penicillin treatment. AB - A 31-year-old woman manifested significant emotional symptoms and personality change due to neurosyphilis; her clinical symptoms were improved with quantitative penicillin treatment. On admission, magnetic resonance imaging (MRI) showed no abnormal findings, while N-isopropyl-p-[123I] iodoamphetamine single photon emission computed tomography (123I-IMP SPECT) initially showed a remarkable increase in cerebral blood flow (CBF) in the cerebral cortex. This increase disappeared after improvement of the patient's clinical symptoms with treatment. It appears that the increase in CBF might have reflected an active inflammatory state of neurosyphilis and that its disappearance might therefore represent successful treatment with penicillin for neurosyphilis. Our case study suggests that single photon emission computed tomography (SPECT) is a useful method for evaluating an inflammatory state and for assessing the effect of therapy on neurosyphilis. PMID- 14751445 TI - Effects of NMDA receptor antagonists on olfactory learning and memory in the honeybee (Apis mellifera). AB - In contrast to vertebrates the involvement of glutamate and N-methyl-D-aspartate (NMDA) receptors in brain functions in insects is both poorly understood and somewhat controversial. Here, we have examined the behavioural effects of two noncompetitive NMDA receptor antagonists, memantine (low affinity) and MK-801 (high affinity), on learning and memory in honeybees (Apis mellifera) using the olfactory conditioning of the proboscis extension reflex (PER). We induced memory deficit by injecting harnessed individuals with a glutamate transporter inhibitor, L-trans-2,4-PDC (L-trans-2,4-pyrrolidine dicarboxylate), that impairs long-term (24 h), but not short-term (1 h), memory in honeybees. We show that L trans-2,4-PDC-induced amnesia is 'rescued' by memantine injected either before training, or before testing, suggesting that memantine restores memory recall rather than memory formation or storage. When injected alone memantine has a mild facilitating effect on memory. The effects of MK-801 are similar to those of L trans-2,4-PDC. Both pretraining and pretesting injections lead to an impairment of long-term (24 h) memory, but have no effect on short-term (1 h) memory of an olfactory task. The implications of our results for memory processes in the honeybee are discussed. PMID- 14751446 TI - Discriminative stimulus properties of 1.25 and 5.0 mg/kg doses of clozapine in rats: examination of the role of dopamine, serotonin, and muscarinic receptor mechanisms. AB - Clozapine (CLZ), an atypical antipsychotic drug (APD), produces minimal extrapyramidal side effects (EPS) and has significant advantages for treating both positive and negative symptoms in schizophrenic patients. CLZ has been established as a discriminative cue in the drug discrimination paradigm and in generalization tests the CLZ cue is more selective for atypical, rather than typical, APDs. However, greater selectivity for atypical antipsychotics has been demonstrated with a lower (1.25 mg/kg) CLZ training dose in rats [Psychopharmacology, 149 (2000) 189], rather than the traditional, higher training dose (5.0 mg/kg). It is therefore of interest to evaluate the properties mediating the 1.25 mg/kg CLZ discriminative cue. In the present study, rats were trained to discriminate either 1.25 mg/kg (N=7) or 5.0 mg/kg (N=7) CLZ from vehicle in a two-lever drug discrimination task. The typical antipsychotic haloperidol (0.1-0.4 mg/kg) did not substitute for either CLZ cue, whereas the atypical antipsychotic melperone (0.37-3.0 mg/kg) provided full substitution in both groups (>80% CLZ-appropriate responding). The 5-HT(1A) receptor agonist (+) 8-OH-DPAT (0.04-0.16 mg/kg), and the selective 5-HT(2A) receptor antagonist M100907 (0.03-1.0 mg/kg) did not produce substitution in either group. (+)-8-OH DPAT combined with haloperidol (0.05 mg/kg) engendered only partial substitution (>60% CLZ-appropriate responding) for both CLZ cues, and M100907 combined with haloperidol (0.05 and 0.1 mg/kg doses) failed to provide substitution in either group. Trihexyphenidyl (0.18-6.0 mg/kg), a muscarinic M(1)-preferring receptor antagonist, engendered full substitution for the 1.25 mg/kg CLZ cue, but only partial substitution for the 5.0 mg/kg CLZ cue. These results provide evidence that antagonism at the muscarinic M(1) receptor is sufficient to provide 1.25 mg/kg CLZ-like discriminative stimulus effects. PMID- 14751447 TI - Exercise, antidepressant treatment, and BDNF mRNA expression in the aging brain. AB - Principal mental disorders affecting the geriatric population include dementia and depression. A lack of trophic support is thought to contribute to the pathology of these disorders. Physical activity and antidepressant treatment increase the expression of brain-derived neurotrophic factor (BDNF) in the young rat hippocampus. Herein, we investigated the responsiveness of the aging rat hippocampus to antidepressant treatment and voluntary exercise. In situ hybridization revealed that, in young animals, exercise, antidepressant treatment, or their combination elevated BDNF mRNA levels in several hippocampal regions, most notably in the CA3, CA4, and dentate gyrus (DG). This effect was rapid (detectable at 2 days) and sustainable to 20 days. In aged (22-month-old) rats, hippocampal responsiveness to antidepressant treatment and exercise was also rapid and sustainable, but evident mostly in the CA1 and CA2. Daily swimming also revealed that small amounts of activity led to marked elevations in hippocampal BDNF mRNA. The differences in regional patterns of BDNF mRNA elevations between young and aged animals observed with running were maintained with this different exercise modality. Our results indicate that the aged brain is responsive to exercise and antidepressant treatment, and changes in regional response patterns may reflect shifts in hippocampal physiology during the lifespan. PMID- 14751448 TI - The chick separation stress paradigm: a validation study. AB - To expand the generalizability of the chick separation stress paradigm as a high throughput anxiolytic screen, six positive drug probes (doses in mg/kg: meprobamate 15-120, pentobarbital 2.5-20.0, chlordiazepoxide 2.5-15.0, buspirone 2.5-10.0, imipramine 1-15, and clonidine 0.10-0.25) and five negative drug probes (amphetamine 0.5-4.0, scopolamine 0.2-1.6, caffeine 5-20, chlorpromazine 1-30, and haloperidol 0.03-1.00) were evaluated in the test. Seven-day-old chicks received intramuscular injections of either vehicle or drug probe 15 min prior to tests in either a mirror (low-stress) or a no-mirror (high-stress) condition for a 3-min observation period. The dependent measures were distress vocalizations to index separation stress and sleep onset latency to index sedation. All positive drug probes attenuated distress vocalizations in a dose-dependent manner, except buspirone. All positive drug probes affected sleep onset latency in a dose dependent manner, except buspirone and imipramine. In all cases, the anxiolytic like effect of positive drug probes was greater than its sedative effect. None of the negative drug probes affected either distress vocalizations or sleep onset latency, except for the highest dose of amphetamine, which caused pronounced stereotypy. These findings demonstrate that this anxiolytic screen is sensitive to a wide range of positive pharmacological probes and insensitive to a wide range of negative pharmacological probes. PMID- 14751449 TI - Delta sleep-inducing peptide and its tetrapeptide analogue alleviate severity of metaphit seizures. AB - The effects of delta sleep-inducing peptide (DSIP) and its tetrapeptide analogue, DSIP(1-4), on metaphit-induced audiogenic seizures were studied. Five groups of adult male Wistar rats were intraperitoneally treated with (1) saline, (2) metaphit, (3) DSIP, (4) metaphit+DSIP and (5) metaphit+DSIP(1-4). To examine blocking effects of DSIP and its analogue on fully developed metaphit seizures, the last two groups were injected after the eight audiogenic testing. The rats were stimulated using electric bell (on the top of the cage, generating 100+/-3 dB and frequency 5-8 kHz, for 60 s) 1 h after metaphit and afterwards at hourly intervals during the experiment. For EEG recordings and power spectra, three gold plated screws were implanted into the skull. In metaphit-treated animals, EEGs appeared as polyspikes and spike-wave complexes while the power spectra were increasing for 30-h period. The incidence and severity of metaphit-induced audiogenic seizures reached peak value 7-12 h after the injection. Both DSIP and DSIP(1-4) significantly increased power spectra of delta waves and decreased incidence of seizures, mean seizure grade and tonic component of metaphit-induced convulsions. Taken together, these results suggest that DSIP and its analogue DSIP(1-4) should be considered as potential antiepileptics. PMID- 14751450 TI - Effects of morphine and cocaine in mice with stable high aggressive and nonaggressive behavioral strategy. AB - The social group experience of mice with opposite aggressive and nonaggressive behavioral strategies was examined to modulate reinforcing effects of morphine and cocaine. Highly aggressive and nonaggressive male mice cohoused for long period in three-member groups were tested to self-administrate the drugs and to develop conditioned place preferring by them. Mouse triads formed by principle of descending aggression were used as a model of linear hierarchical group. The level of mouse aggression was identified previously within the stock group and during encounter with unknown intruder that continued to be stable over the time of experiment. Highly aggressive mice self-administered morphine and cocaine at higher unit concentrations (1.5 and 1.5 mg/ml) as compare with nonaggressive animals (0.5 and 0.25, 0.5, 1.0 mg/ml). Both morphine (2.5, 5.0, 10.0, and 20.0 mg/kg) and cocaine (2.5, 5.0, and 10.0 mg/kg) induced conditioned place preference in nonaggressive mice at all doses. In contrast, morphine had no effect in highly aggressive mice, while cocaine induced place conditioning at the highest doses (10 mg/kg) only. Our results illustrate that social experience in a stable group alter mouse sensitivity to the rewarding properties of drugs of abuse and social state should be taken into account in the experiments when social interactions are present. PMID- 14751451 TI - Behavioral effects of manganese injected in the rat substantia nigra are potentiated by dicumarol, a DT-diaphorase inhibitor. AB - The purpose of this study was to evaluate the contribution of DT-diaphorase inhibition to in vivo neurodegenerative effects of dopamine (DA) oxidation to the corresponding o-quinones. The neurotoxicity to nigrostriatal DA neurons was induced by injection of manganese pyrophosphate (Mn(3+)) complex as a prooxidizing agent alone or together with the DT-diaphorase inhibitor dicumarol into the right rat substantia nigra. The behavioral effects were compared with those induced after selective lesions of dopaminergic neurons with 6 hydroxydopamine (6-OHDA). Intranigral injection of Mn(3+) and Mn(3+) plus dicumarol produced significant impairment in motor behavior compared with control animals. However, the effect seen in the Mn(3+) plus dicumarol injected group was significantly more severe than that observed in the Mn(3+) alone injected group. In motor activity and rearing behavior, the simultaneous injection of Mn(3+) plus dicumarol produced a 6-OHDA-like impairment. Similar effects were observed in the acquisition of a conditioned avoidance response (CAR). Dicumarol significantly impaired avoidance conditioning although without affecting the motor behavior. The behavioral effects were correlated to the extent of striatal tyrosine hydroxylase (TH)-positive fiber loss. Rats receiving unilateral intranigral Mn(3+) and Mn(3+) plus dicumarol injections exhibited a significant reduction in nigrostriatal TH-positive fiber density in medial forebrain bundle compared with the contralateral noninjected side. In conclusion, this study provides evidence that the neurotoxicity of Mn(3+) in vivo is potentiated by DT-diaphorase inhibition, suggesting that this enzyme could play a neuroprotective role in the nigrostriatal DA systems. PMID- 14751453 TI - Precipitated opioid withdrawal across acute physical dependence induction methods. AB - The phenomenon of acute opioid physical dependence (APD) is an established and well-characterized experimental model for studying the clinical phenomenon of physical dependence to opioids in humans. In this paradigm, an opioid withdrawal syndrome is elicited in non-opioid-dependent humans by the parenteral administration of naloxone (NX) following a single large dose of opioid agonist. Although induced by various opioids and NX administration schedules, lacking is a direct comparison of different induction protocols with respect to withdrawal severity. Using a crossover design, we compared withdrawal severity in four healthy male subjects pretreated with morphine (MS; 18 mg/70 kg im), MS (10 mg/70 kg iv) and hydromorphone (HM; 2 mg/70 kg) followed 2 or 6 h later with NX (10 mg/70 kg iv). Dependent measures included both physiological and subjective indicators of withdrawal. All opioid pretreatments reliably induced APD and repeated-measures analysis of variance (ANOVA) showed that both pattern and severity of precipitated withdrawal were similar across conditions. Thus, despite altering the pretreatment opioid and route of administration, all three APD protocols produced similar and reliable withdrawal symptoms in humans. PMID- 14751452 TI - Co-exposure to pyridostigmine bromide, DEET, and/or permethrin causes sensorimotor deficit and alterations in brain acetylcholinesterase activity. AB - Military personnel deployed in the Persian Gulf War (PGW) were exposed to a combination of chemicals, including pyridostigmine bromide (PB), DEET, and permethrin. We investigated the dose-response effects of these chemicals, alone or in combination, on the sensorimotor performance and cholinergic system of male Sprague-Dawley rats. Animals were treated with a daily dermal dose of DEET and/or permethrin for 60 days and/or PB (gavage) during the last 15 days. Neurobehavioral performance was assessed on day 60 following the beginning of the treatment with DEET and permethrin. The rats were sacrificed 24 h after the last treatment for biochemical evaluations. PB alone, or in combination with DEET, or DEET and permethrin resulted in deficits in beam-walk score and longer beam-walk times compared to controls. PB alone, or in combination with DEET, permethrin, or DEET and permethrin caused impairment in incline plane performance and forepaw grip strength. PB alone at all doses slightly inhibited plasma butyrylcholinesterase activity, whereas combination of PB with DEET or permethrin increased its activity. Brainstem acetylcholinesterase (AChE) activity significantly increased following treatment with combinations of either DEET or permethrin at all doses, whereas the cerebellum showed a significant increase in AChE activity following treatment with a combination of PB/DEET/permethrin. Co exposure to PB, DEET, and permethrin resulted in significant inhibition in AChE in midbrain. PB alone or in combination with DEET and permethrin at all doses increased ligand binding for m2 muscarinic acetylcholine receptor in the cortex. In addition, PB and DEET together or a combination of PB, DEET, and permethrin significantly increased ligand binding for nicotinic acetylcholine receptor. These results suggest that exposure to various doses of PB, alone and in combination with DEET and permethrin, leads to sensorimotor deficits and differential alterations of the cholinergic system in the CNS. PMID- 14751454 TI - Acute stressor-selective effect on total plasma homocysteine concentration in rats. AB - Stress produces several physiological and behavioral alterations that increase cardiovascular morbidity and mortality. Nonetheless, there is a dearth of studies that have evaluated the effects of stress on total plasma homocysteine, an important amino acid associated with cardiovascular disease. We used four distinct acute stressors in rats, i.e., swimming, restrain, novelty and cold exposure, in order to examine whether any acute effect on total plasma homocysteine concentrations would occur. Plasma corticosterone and adrenocorticotropic hormone concentrations were also measured to demonstrate the ability of the chosen manipulations to activate the hypothalamic-pituitary adrenal (HPA) axis. Three of the four stressors activated the HPA axis and only restrain affected total plasma homocysteine concentrations (+37%, P=.006) compared with the control group. The complexity of the physiological responses to stress, the peculiarities of stress responses and the intricate regulatory systems involved in homocysteine metabolism must be taken into account in order to clarify the increasing effect of restrain (mainly a psychological stressor) on total plasma homocysteine concentrations in rats and to evaluate its meaning in human pathology. PMID- 14751455 TI - Involvement of melatonin metabolites in the long-term inhibitory effect of the hormone on rat spinal nociceptive transmission. AB - There is evidence that melatonin and its metabolites could bind to nuclear sites in neurones, suggesting that this hormone is able to exert long-term functional effects in the central nervous system via genomic mechanisms. This study was designed to investigate (i) whether systemically administered melatonin can exert long-term effects on spinal cord windup activity, and (ii) whether blockade of melatonin degradation with eserine could prevent this effect. Rats receiving melatonin (10 mg/kg ip), the same dose of melatonin plus eserine (0.5 mg/kg ip), or saline were studied. Seven days after administration of the drugs or saline, spinal windup of rats was assessed in a C-fiber reflex response paradigm. Results show that rats receiving melatonin exhibited a reduction in spinal windup activity. This was not observed in the animals receiving melatonin plus eserine or saline, suggesting a role for melatonin metabolites in long-term changes of nociceptive transmission in the rat spinal cord. PMID- 14751456 TI - Reduced 5-HT3 receptor binding and lower baseline plus maze anxiety in the alcohol-preferring inbred fawn-hooded rat. AB - The present investigation sought to explore the relationship between the 5-HT(3) receptor and anxiety-like behavior in fawn-hooded (FH/Wjd) rats, an inbred strain that exhibits a high intake and preference for ethanol, and the alcohol nonpreferring ACI/N strain. Using quantitative autoradiography, we examined whether there were differences in central 5-HT(3) receptor binding in FH/Wjd versus ACI/N rats. Ten to 14 days prior to being used in the autoradiographic studies, rats were first confirmed to be representative of their strains by subjecting them to a two-bottle choice procedure for 2 weeks. The binding of [3H]LY 278584 to 5-HT(3) receptors was significantly reduced in frontal cortex, CA1 region of hippocampus, and in the medial and lateral nuclei of the amygdala of FH/Wjd versus ACI/N rats. In the anterior cingulate cortex and in the dentate gyrus region of the hippocampus the reduction in [3H]LY 278548 binding in the FH/Wjd versus ACI/N strain (40% and 41%, respectively) did not reach statistical significance. In a separate group of animals, the effects of the 5-HT(3) receptor antagonist MDL 72222 (3 mg/kg ip) on anxiety-related behaviors were assessed in the elevated plus maze. In vehicle-treated rats, the FH/Wjd strain exhibited significantly greater percent of time spent on the open arms and percent open arm entries, an indication of less anxiety. Pretreatment with MDL 72222 did not alter these behaviors in the FH/Wjd rats, but had an anxiolytic-like effect in the ACI/N strain, significantly increasing the percent of time spent on the open arms and percent open arm entries. Further research into 5-HT(3) receptor function in the alcohol-preferring FH/Wjd rats is needed to elucidate the relationship among 5-HT(3) receptors, alcohol drinking, and anxiety. PMID- 14751458 TI - Antidepressant-like effect of nicotinamide adenine dinucleotide in the forced swim test in rats. AB - Nicotinamide adenine dinucleotide (NADH), a cosubstrate for energy transfer in the oxidative phosphorylation, has supposedly beneficial effects on central nervous system (CNS)-related diseases, e.g., shown in an open study with depressive patients. To our knowledge there are no data concerning the efficacy of NADH in animal tests. Acute effects of NADH and the precursor nicotinamide, compared to controls and the antidepressants desipramine and fluoxetine, were examined in the forced swim test (FST) in Wistar rats. NADH, but not nicotinamide, reduced immobility and increased swimming behaviour in the FST, with a minimum effective dose of 5 mg/kg. NADH-induced behavioural profile was similar to fluoxetine, but different from desipramine. Since NADH did not induce hyperlocomotion but even decreased motor activity in the open field test, the antidepressant-like effect cannot be attributed to an increase in motor activity. These data support an antidepressant potential of NADH. PMID- 14751457 TI - Sensitivity to drug effects on prepulse inhibition in inbred and outbred rat strains. AB - Genetic differences in the neurochemical regulation of PPI in rats may help clarify the neural basis of inherited PPI differences in neuropsychiatric disorders. We reported and characterized substantial heritable differences in sensitivity to PPI-disruptive effects of DA agonists in outbred Sprague Dawley (SDH) versus Long-Evans (LEH) rats. Other strains might yield large group separations and facilitate studies of the neural basis for these strain differences; inbred strains might also allow us to map genes associated with differential PPI sensitivity. Sensitivity to the PPI-disruptive effects of the DA agonist apomorphine (APO) and the NMDA antagonist phencyclidine (PCP) were compared across inbred and outbred strains. APO sensitivity was greatest in SDH and buffalo rats, but the effect in buffalo rats was complicated by significant APO-induced startle suppression. PPI APO sensitivity was least in ACI and LEH rats; F344s exhibited intermediate sensitivity and Lewis rats showed a nonlinear dose response (sensitivity at low but not higher doses). PPI APO insensitivity in ACI rats developed over time, with ACI pups exhibiting robust sensitivity. Substantial strain differences were observed in short-interval (10-30 ms) prepulse effects, and APO-induced increases in short-interval PPI occurred in SDH, LEH, and Lewis rats, but not in F344, ACI, or buffalo rats. Sensitivity to PPI-disruptive effects of PCP was generally greater in outbred than inbred rats. These findings identify strains suitable for comparisons of PPI neural circuitry and others for whom such comparisons would be complex and perhaps less informative. PMID- 14751459 TI - Modulation of the locomotor activating effects of the noncompetitive NMDA receptor antagonist MK801 by dopamine D2/3 receptor agonists in mice. AB - The noncompetitive NMDA receptor antagonist MK801 (dizocilpine) produces behavioral stimulation mediated, in part, through indirect activation of the dopamine (DA) system. Previous reports indicate that D2/3 agonists inhibit MK801 induced stereotypies; however, it is unclear if these agonists also attenuate MK801-induced locomotion. As such, the ability of the D2/3 agonists, quinelorane and quinpirole, and the partial D3 agonist, BP897, to attenuate the locomotor activating effects of MK801 was examined in mice. MK801 (0.1-1.0 mg/kg) produced a biphasic effect on total distance traveled with the intermediate dose of 0.3 mg/kg producing the greatest stimulation. The increase in MK801-induced total distance traveled was attenuated by the coadministration of quinelorane and quinpirole at doses that alone had no effect on activity. Similarly, the partial D3 agonist, BP897, blocked the effects of MK801. The D3-preferring antagonist, nafadotride, reversed the attenuation of quinelorane and partially reversed the attenuation of quinpirole. The D2-preferring antagonist, eticlopride, reversed the attenuating effects of quinelorane, but was not effective against quinpirole. Nafadotride and eticlopride were ineffective against the attenuating effects of BP897 on MK801-induced locomotion. Because BP897 is a partial agonist it was tested against quinelorane/MK801 and quinpirole/MK801 combinations. BP897 reversed the attenuating effects of quinelorane, but not those of quinpirole on MK801's effects. These results demonstrate that the DA system, through D2/3 receptor activation, modulates the locomotor activating effects produced by noncompetitive NMDA receptor blockade. PMID- 14751460 TI - Chronic methamphetamine increases fighting in mice. AB - A propensity for violent behaviors to develop in chronic methamphetamine (METH) abusers has been noted. The idea that increased aggressiveness might result from chronic METH administration was tested in mice after chronic (long-term intermittent, 8 weeks) or single exposures to the drug. A single injection of METH (6 mg/kg) did not augment fighting. In contrast, chronic METH administration significantly increased the number of animals that initiated bite attacks. This regimen also shortened the latency before the first attack. Latency before the first attack was shorter at 20 h after the METH injection than at 15 min after injection. Locomotor activity was not different at 20 h after METH injection, indicating that increased fighting was not secondary to METH-induced hyperactivity. METH-induced increases in fighting were not related to the duration of persistent sniffing after the initial encounter with an intruder since the duration of this behavior was significantly increased at 15 min after METH but not at 20 h post drug. These results indicate that repeated injections of METH can increase fighting behaviors and also alter social interactions in mice. Thus, intermittent administration of METH might be useful as a pharmacological model to study the biochemical and molecular bases of aggressiveness. PMID- 14751461 TI - Long-term opiate effects on amphetamine-induced dopamine release in the nucleus accumbens core and conditioned place preference. AB - Withdrawal following chronic exposure to opiates or other drugs of abuse, administered as frequent doses, or a chronic infusion can cause reductions in mesolimbic dopamine (DA) transmission. However, mesolimbic DA transmission can be enhanced by opiates or psychostimulants administered intermittently as a single daily injection. Both enhanced and attenuated responsiveness of the mesolimbic DA system may have important implications for substance abuse disorders. Previous studies have shown that procedures that use electrical stimulation or drug treatments to augment neurotransmitter release are more effective for demonstrating declines in mesolimbic DA transmission that persist for extended periods following opiate withdrawal. The present study evaluated the effects of pretreatment with noncontingent morphine on amphetamine-induced DA release in the nucleus accumbens core and conditioned place preference (CPP). Morphine pretreatment was administered as a constant infusion, which was gradually increased to a dose of 50 mg/kg/day over a 1-week period in Wistar rats. At 10 days after cessation of morphine pretreatment, baseline dialysate DA levels in the nucleus accumbens core were unchanged, but amphetamine-induced increases in DA were attenuated by greater than 50% in morphine-pretreated animals. Morphine pretreatment did not modify locomotor activity during conditioning sessions, expressed as absolute values or change in activity counts between saline and morphine injections. Place preference, conditioned by two morphine pairings at 10 and 11 days after the onset of opiate withdrawal, was enhanced by opiate pretreatment between 12 and 33 days after the onset of withdrawal. In conclusion, morphine pretreatment delivered as a constant infusion can have pronounced and long-lasting effects on DA release and CPP, which may have important implications for drug-seeking behavior and treatment of substance abuse disorders. PMID- 14751462 TI - Antagonism of anticholinesterase (DFP) toxicity by donepezil plus scopolamine: a preliminary study. AB - Studies in animals exploring the antagonism of the cholinesterase inhibitors soman and sarin have shown that pretreatment with low doses of the centrally acting cholinesterase inhibitor, physostigmine, alone or in conjunction with the centrally acting anticholinergic agent, scopolamine, is effective against their lethality and toxicity. The current study evaluated the effects of pretreatment with the oral anticholinesterase agent, donepezil (Aricept, 2.0 mg/kg), used to treat Alzheimer's disease, with and without scopolamine in decreasing the hypothermic, hypokinetic, and diarrhea-inducing effects of the irreversible long acting cholinesterase inhibitor diisopropyl fluorophosphate (DFP, 1.0 mg/kg) in adult Flinders sensitive line (FSL) male rats. Donepezil alone and donepezil plus scopolamine (0.1 mg/kg) to a greater extent antagonized the decrease in temperature, hypoactivity, and induction of diarrhea due to DFP observed at 4 h after its administration. However, donepezil alone induced hypothermia at 1 and 2 h after treatment. Therefore, these preliminary findings are encouraging, but many additional studies are needed to establish the effectiveness of donepezil as a prophylactic agent against irreversible cholinesterase inhibition by DFP. PMID- 14751463 TI - Anticonvulsive effect of agmatine in mice. AB - The present study was designed to examine the effect of agmatine, the decarboxylated product of L-arginine by L-arginine decarboxylase, on convulsion in the mouse maximal electroshock (MES) test and mouse glutamate-induced convulsant test. MES convulsion and glutamate convulsion were respectively induced by an electrical stimulation (110 V, 0.3 s, 8 Hz) and by intracerebroventricular injection of glutamate (0.5 M, pH 7.4, 5microl). The results were expressed as the tonic and clonic time of convulsion in MES or percentage of mice with tonic hind-limb extension in glutamate-induced convulsant assay. Agmatine given intracerebroventricularly (2-16 mg/kg) or subcutaneously (10-160 mg/kg) significantly shortened the tonic and clonic times of convulsion in a dose-dependent manner in the mouse MES test. Glutamate (0.5 M, 5microl icv per mouse) induced an obvious convulsive response indicated by tonic hind-limb extension in mice, and agmatine (2-16 mg/kg icv) decreased the rate of mice with tonic hind-limb extension like NMDA receptor antagonist MK-801. The anticonvulsive effect of agmatine (80 mg/kg sc) on both the tonic and clonic times of convulsion lasted for more than 4 h after administration in the mouse MES test, which was twice that of barbital. Taken together, the results implicate that agmatine has obvious anticonvulsive effects, and its possible mechanism might be related to the antagonism of the function of NMDA receptors. PMID- 14751464 TI - Fenfluramine-induced hypothermia is associated with cutaneous dilation in conscious rats. AB - The antiobesity agent, fenfluramine, produces hypothermia in rodents by an, as yet, uncharacterized mechanism. The present study was conducted in conscious rats to determine if fenfluramine-induced hypothermia was associated with cutaneous dilation. In animals maintained at 16 degrees C, core body temperature (T(CORE)) was measured telemetrically, and tail surface temperature was monitored with thermocouples fixed to the tail (T(TAIL)). D-Fenfluramine (10 mg/kg ip) produced a rapid increase in T(TAIL) of 7.7+/-0.4 degrees C (P<.001) and a decline in T(CORE) of 4+/-0.3 degrees C (P<.001). Two findings indicate that the increase in T(TAIL) was due to the withdrawal of a sympathetic vasoconstrictor tone. First, pretreatment with the ganglionic blocker, pentolinium, prevented fenfluramine induced changes in T(TAIL). Second, when sympathetic tone to the tail was physiologically withdrawn by increasing the environmental temperature to 28 degrees C, fenfluramine treatment produced no increase in T(TAIL). Moreover, the effects of fenfluramine on T(TAIL) and T(CORE) depended on the uptake of fenfluramine into serotonergic neurons because these effects were markedly attenuated by pretreatment with the selective serotonin re-uptake inhibitor, fluoxetine. The hypothermic effect of fenfluramine occurred despite the fact that total body oxygen consumption increased by 20%. The results suggest that heat loss due to the dilation of the cutaneous circulation contributes to fenfluramine induced hypothermia. PMID- 14751465 TI - Anxiolytic-like effects of rose oil inhalation on the elevated plus-maze test in rats. AB - The effect of rose oil inhalation (1.0%, 2.5%, and 5.0% w/w) on the elevated plus maze (EPM) test was investigated in adult male rats and compared with the effect of diazepam (DZP) (1.0 and 2.0 mg/kg) administered intraperitoneally 30 min before testing. Exposure to rose oil produced an anxiolytic-like effect similar to DZP (anxiolytic reference drug). Thus, at some concentrations, rose oil significantly increased the number of visits to and time spent in the open arms of the EPM. Anxiolytic-like properties of rose oil were observed using the EPM, being consistent with other behavioral and clinical studies. PMID- 14751466 TI - Physiological and behavioral effects of methamphetamine in a mouse model of endotoxemia: a preliminary study. AB - We investigated the effects of methamphetamine (METH) on core body temperature (Tb) and motor activity (MA) with or without exposure to a peripheral immune challenge. Mice were exposed to an escalating METH treatment and then to a METH treatment known to cause neurotoxicity (binge METH treatment). This was followed by a challenge with lipopolysaccharide (LPS). Three days later, METH and saline treated control groups were challenged with an acute test dose of METH (METH test). Animals exposed to the escalating METH treatment exhibited a significant increase in Tb only after the initial exposure to METH (Day 1) and following the METH test (Day 7). The hyperthermic effect produced by the METH test (Day 7) was reduced in mice previously exposed to combined exposure to binge METH and LPS treatments. The escalating METH treatment produced MA sensitization to the METH test. Animals treated with the binge METH, LPS injection or both treatments combined prevented MA sensitization to the METH test. These findings suggest that induction of peripheral endotoxemia in animals with a history of METH reduced the hyperthermic response to a subsequent challenge with METH. PMID- 14751467 TI - Long-term social isolation and medial prefrontal cortex: dopaminergic and cholinergic neurotransmission. AB - Rearing rats in social isolation has been suggested as an animal model of schizophrenia, based mainly on the similarity between the attenuation of prepulse inhibition (PPI) in isolated rats and in schizophrenic patients. The medial prefrontal cortex (mPFC) plays a major role in the pathophysiology of schizophrenia. Thus, a postmortem micropunch analysis measuring dopamine (DA), DOPAC (3,4-dihydroxyphenylacetic acid) and homovanillic acid (HVA) in the dorsal and ventral subregion of the mPFC, the caudate putamen (CPu) and nucleus accumbens (NAc) was carried out on socially isolated or group-housed male Sprague Dawley (SD) rats. Additionally, in vivo microdialysis with D-amphetamine (1 mg/kg ip) stimulation was performed in isolated animals and their controls, examining the ventral mPFC for acetylcholine (ACh), DOPAC and HVA levels. Simultaneously, recording of motor activity was performed. In the neurochemical postmortem tissue analysis we found no difference in any of the brain regions tested between isolated and group-reared animals. Amphetamine increased ACh levels in the mPFC, induced a decrease in DOPAC and HVA levels, and increased motor activity. A close to significant Drug x Housing interaction reflected the fact that the amphetamine induced decrease of DOPAC was confined to the group-housed animals. In conclusion, social isolation leads only to moderate changes in the dopaminergic system in the mPFC, whereas the cholinergic system remains unaffected. PMID- 14751468 TI - Erythropoietin improves place learning in fimbria-fornix-transected rats and modifies the search pattern of normal rats. AB - The acquisition of a water-maze-based allocentric place learning task was studied in four groups of rats: two groups subjected to bilateral transections of the fimbria-fornix and two groups undergoing a sham control operation. At the moment of surgery all animals were given one systemic (intraperitoneal) injection of either human recombinant erythropoietin (EPO) (at a dosage of 5000 IU/kg body weight), given to one of the fimbria-fornix-transected groups and one of the sham operated groups, or vehicle (saline), given to the two remaining groups. The 25 day task acquisition period (one session/day) began 6 or 7 days after the day of surgery. The fimbria-fornix-transected and saline-injected group exhibited a pronounced and long-lasting impairment of task acquisition. In contrast, the fimbria-fornix-transected and EPO-treated group demonstrated a less pronounced and more transient lesion-associated impairment. The two sham-operated groups did not differ with respect to the proficiency of task acquisition. But administration of EPO to intact animals caused a significant modification of swim patterns-apparently reflecting a somewhat modified strategy of task solution. It is concluded that systemic administration of EPO significantly improves the posttraumatic functional recovery of the presently studied place learning task after transections of the fimbria-fornix. Additionally, administration of EPO influences the strategy, although not quality, of task solution in normal (sham operated) rats. PMID- 14751469 TI - R-citalopram inhibits functional and 5-HTP-evoked behavioural responses to the SSRI, escitalopram. AB - Escitalopram mediates the serotonin re-uptake inhibitory and antidepressant effect of citalopram racemate. However, recent studies have shown that R citalopram inhibits the escitalopram-induced increase of extracellular 5-HT levels in the frontal cortex of rats. Here, we investigated the inhibitory effect of R-citalopram on the escitalopram-induced increase of 5-HT neurotransmission at the behavioural [potentiation of 5-hydroxytryptophan (5-HTP)-induced behavioural changes in mice and rats] and functional (increase in serum corticosterone in rats) levels. The effect of escitalopram was inhibited by R-citalopram in all three models, and R-citalopram, given alone, was inactive. The effects were more pronounced using an escitalopram to R-citalopram ratio of 1:4 than ratios of 1:2 and 1:1, suggesting a dose-dependent effect. The ED(50)-value of escitalopram in mouse 5-HTP potentiation studies corresponded to a serum concentration of approximately 50 ng/ml, which can be considered to be in the range of clinically relevant serum concentrations. In conclusion, R-citalopram inhibited the escitalopram-induced increase of 5-HT activity in functional, as well as behavioural, animal models. The mechanism involved in this interaction is currently unknown, but may be related to an improved clinical effect seen with escitalopram in comparison with citalopram. PMID- 14751470 TI - Sedative and sleep-enhancing properties of linarin, a flavonoid-isolated from Valeriana officinalis. AB - We have recently reported the presence of the anxiolytic flavone 6-methylapigenin (MA) and of the sedative and sleep-enhancing flavanone glycoside 2S (-) hesperidin (HN) in Valeriana officinalis and Valeriana wallichii. MA, in turn, was able to potentiate the sleep-inducing properties of HN. The present paper reports the identification in V. officinalis of the flavone glycoside linarin (LN) and the discovery that it has, like HN, sedative and sleep-enhancing properties that are potentiated by simultaneous administration of valerenic acid (VA). These effects should be taken into account when considering the pharmacological actions of valeriana extracts. PMID- 14751472 TI - Insertion of the LMA-Unique with and without digital intraoral manipulation by inexperienced personnel after manikin-only training. AB - In a randomized, crossover study, we compared insertion of the LMA-Unique with and without digital intraoral manipulation by inexperienced personnel after manikin-only training. Ten registered nurses with no hands-on clinical experience of airway management and 100 anesthetized, paralyzed adults (ASA Status 1-2, aged 18-80 years) participated in the trial. Training comprised: 1) a 30-min didactic lecture; 2) a 3-min description of each of the insertion techniques; 3) a 3-min demonstration of each technique using a manikin; 4) 10 min of supervised training on the manikin with each technique. The time to achieve an effective airway (2 consecutive expired tidal volumes >/= 8 ml/kg; maximum 90 s or 3 attempts allowed) and the number of insertion attempts were determined by analysis of digital video recordings. Any blood staining on the first randomized device was noted at removal. The first attempt success rate (with digital intraoral manipulation, 84%; without digital intraoral manipulation, 87%) and overall success rate (with digital intraoral manipulation, 94%; without digital intraoral manipulation, 93%) were similar. Effective airway time was shorter without digital intraoral manipulation (43 +/- 17 vs. 33 +/- 15 s). Blood staining was detected in 12% (6/50) with and 16% (8/50) without digital intraoral manipulation (not significant). There were no differences in performance among nurses. We conclude that insertion of the LMA-Unique is equally successful with or without digital intraoral manipulation by inexperienced personnel in paralyzed adults after manikin-only training. Successful insertion of the LMA-Unique does not require insertion of the finger into the patient's mouth. PMID- 14751471 TI - Modulation of multiple ethanol withdrawal-induced anxiety-like behavior by CRF and CRF1 receptors. AB - Previous work demonstrated that rats subjected to multiple withdrawals from chronic ethanol exhibit a sensitization of anxiety-like behavior compared to animals withdrawn from treatment with an equal but continuous amount of ethanol. This study sought to examine whether corticotropin-releasing factor (CRF) could modulate this ethanol-withdrawal-induced anxiety-like behavior. Initially, rats were administered with CRF (1 microg) or vehicle intraventricularly on two occasions 5 days apart while on control diet (CD) followed by exposure to 7% ethanol diet (ED) for 5 days, with social interaction assessed 5 h into withdrawal. Social interaction was significantly reduced in the CRF-treated animals compared to vehicle-treated rats and vehicle- and CRF-treated rats maintained on CD, indicative that CRF given before ethanol exposure was capable of inducing an adaptive change that sensitized withdrawal-induced anxiety-like behavior. Next, the CRF(1) receptor antagonist CRA1000 (3 mg/kg, systemically), the CRF(2) receptor antagonist antisauvagine-30 (20 microg intraventricularly), or vehicle was injected 4 h after the ethanol was removed following the first and second cycles of chronic ethanol exposure and the effect on the multiple withdrawal-induced anxiety-like behavior determined after the third withdrawal cycle. The CRF(1) receptor antagonist blocked the reduced social interaction behavior, whereas the CRF(2) receptor antagonist was without effect. Similar pretreatment with another CRF(1) receptor antagonist CP-154,526 (10 mg/kg systemically) during the first and second withdrawals also counteracted anxiety like behavior. These findings indicate that the CRF system and CRF(1) receptors play key roles in the adaptive change responsible for the anxiety-like behavior induced by repeated withdrawals from chronic ethanol. PMID- 14751473 TI - Patient preference for route of pain medication delivery. AB - We sought to determine Emergency Department (ED) patient preference for oral (p.o.), intramuscular (i.m.), or intravenous (i.v.) pain medication and patient expectations of time to medication effect by route. A prospective, observational study of 1276 patients presenting with painful illness or injury was performed in a university ED. Patient preferences were 66% p.o., 15% i.m., and 19% i.v. pain medication. Patients aged greater than 55 years were more likely to prefer parenteral medication than younger patients. Patients in severe pain were also more likely to prefer parenteral medication than those with less severe pain. Despite these differences, a majority of patients in all groups preferred oral medications. There were no differences in preference based on ethnicity or gender. Patient expectations for time to pain medication effect were 27 min p.o. (95% CI 26-28), 12 min i.m. (95% CI 11-13), and 7.5 min i.v. (95% CI 6.9-8.0). PMID- 14751474 TI - Patient satisfaction in the Emergency Department: a review of the literature and implications for practice. AB - This article reviews the empirical literature on patient satisfaction in the Emergency Department (ED). It explores the implications for clinical practice, discusses limitations and weaknesses of the literature, and provides direction for future research. Articles resulting from a comprehensive electronic search were obtained, their references examined, and all other relevant articles not already discovered via the electronic search were acquired and reviewed. Articles were included if: 1) the stated goal of the study was to investigate satisfaction with at least one aspect of ED care, 2) the study was conducted in the United States, 3) it provided enough information on the study methods, design, and statistical analyses to conduct a critical review, and 4) it used quantitative methods. Fifty studies met the above criteria. Based on the multivariate predictive studies, the most robust predictor of global satisfaction is the quality of interpersonal interactions with the ED provider. Perceived waiting times are more closely associated with satisfaction than actual waiting times. Several methods for improving satisfaction have shown promise, but none has garnered sufficient support to recommend unequivocally. Promising interventions include: providing information on how the ED functions through visual media, improving ED processes through performance improvement methodologies, and improving the interpersonal skills of providers. Interventions designed to reduce actual waiting times have not been sufficiently studied, but results from several well-designed studies suggest that such a strategy is unlikely to have as great an impact as those targeting perceived waiting times. To advance this area of research, investigators must use: 1) larger, more representative samples; 2) reliable and valid assessment instruments; 3) theory-driven hypothesis testing; and 4) randomized, controlled trials. PMID- 14751475 TI - Behavioral and psychological risk factors for traumatic injury. AB - The objectives of the present study were to determine the behavioral and psychological risk factors associated with injury. The most widely investigated risk factor for injury is alcohol use. However, other behavioral and psychological risk factors may also contribute to injuries. This study examined the association of alcohol use, injury-related risk behaviors, and psychological characteristics with injury status. A hospital-based case control study of 177 patients admitted for treatment of traumatic injury, and 195 general surgery patients as controls, was conducted. Alcohol use, injury-related risk behaviors, impulsivity, sensation seeking, and risk perception were assessed. The results from a multivariate analysis controlling for age, gender, and ethnicity indicated that, in addition to alcohol consumption (OR = 2.2, 95% CI = 1.1-4.5), driving (OR = 2.4, 95% CI = 1.7-3.5) and violence-related risk behaviors (OR = 1.6, 95% CI = 1.0-2.2) are significantly associated with injury. In conclusion, the engagement in injury-related risk behaviors, including alcohol use, is strongly associated with injury status. Brief interventions that directly address these behavioral risk patterns in injured patients may reduce their risk of future injury. PMID- 14751476 TI - Radiograph use in low back pain: a United States Emergency Department database analysis. AB - We identified factors associated with radiograph evaluation for patients who presented to the Emergency Department (ED) with uncomplicated low back pain (LBP). Using 1998-2000 ED data from the National Hospital Ambulatory Medical Care Survey, a multivariate analysis was performed to assess utilization of radiographs for LBP. Based upon published guidelines, of the over 3 million patients who met our criteria of uncomplicated LBP, 17.8% received an unnecessary radiograph. Patients who arrive via ambulance with moderate pain, who need to be seen within 15 min, and who have 3 or more screening tests ordered are 100% likely to also get a radiograph. There is an increased probability of receiving a radiograph for those patients 40-70 years old, being seen at a metropolitan hospital, having private insurance, and being treated by a resident in training. Multiple factors are associated with the overuse of radiographs for patients presenting with uncomplicated LBP. PMID- 14751477 TI - A case series of patients with black tar heroin-associated necrotizing fasciitis. AB - In 2000, a series of cases of necrotizing fasciitis (NF) among injection users of black tar heroin was observed in our Emergency Department (ED). The description of these cases characterizes the ED presentation of NF, focusing on the sensitivity of physical signs and ED diagnostic tests. All cases of pathologically determined NF in 2000 were prospectively and retrospectively identified and analyzed. Among the 20 patients identified, 20% had fever and 80% were tachycardic in the ED. Most patients, 95%, had elevated white blood cell counts (mean = 37.1 + 23.4 k/ul), 79% had elevated lactate levels, 47% were hyponatremic, and 33% had soft tissue gas on radiograph. Thirty-seven percent of blood cultures and 75% of surgical wound cultures were positive, revealing a variety of organisms. Although tachycardia, leukocytosis, and elevated lactate levels are common in NF patients, fever and soft tissue gas are not. Tests and traditional teaching regarding the "classic" findings of NF should be amended to reflect differences seen in the setting of injection drug use with black tar heroin. PMID- 14751478 TI - Spinal epidural abscess presenting as intra-abdominal pathology: a case report and literature review. AB - Spinal epidural abscess is a rare infectious disease. However, if left unrecognized and untreated, the clinical outcome of spinal epidural abscess can be devastating. Correctly diagnosing a spinal epidural abscess in a timely fashion is often difficult, particularly if the clinician does not actively consider the diagnosis. The most common presenting symptoms of spinal epidural abscess include backache, radicular pain, weakness, and sensory deficits. However, early in its course, spinal epidural abscess can also present with vague and nondescript manifestations. In this report, we describe a case of spinal epidural abscess presenting as abdominal pain, and review the literature describing other cases of spinal epidural abscess presenting as intra-abdominal pathology. PMID- 14751479 TI - A subglottic mass mimicking near-fatal asthma: a challenge of diagnosis. AB - Upper airway obstruction due to a subglottic tumor can be easily misdiagnosed as bronchial asthma. We report on a 50-year-old woman who was ultimately diagnosed with subglottic tumor, but who presented with near-fatal asthma. According to her medical history she had been treated with high doses of prednisolone and bronchodilators for the past year for difficult asthma. The patient presented to the Emergency Department (ED) in severe respiratory distress. The chest X-ray study revealed bilateral hyperinflation. The flow-volume curve suggested a fixed airway obstruction. After performing a laryngoscopic examination, a subglottic mass was discovered and an urgent tracheotomy was performed. After the operation, all symptoms and respiratory distress disappeared. This case report emphasizes the fact that not all wheezes are attributable to asthma. Upper airway obstructions can lead to asthma-like symptoms in which establishment of the correct diagnosis may be challenging. PMID- 14751480 TI - Hemotympanums secondary to spontaneous epistaxis in a 7-year-old. AB - Hemotympanum is a well-known physical finding most often associated with basilar skull fractures and therapeutic nasal packing. A literature review demonstrated only five cases of hemotympanum associated with spontaneous epistaxis in adults. To our knowledge, there have been no reported cases in children. We present a case of a 7-year-old child with bilateral hemotympanums secondary to spontaneous epistaxis. PMID- 14751481 TI - Pediatric cardiac disorders. AB - Although pediatric cardiac disorders are not commonly seen in the Emergency Department, they are important to identify to prevent further morbidity or mortality. Diagnosis may be complicated by the lack of classic complaints such as chest pain or palpitations that are commonly associated with cardiac disorders. In fact, presenting complaints associated with pediatric cardiac disorders, like "fussiness" or "difficulty feeding," may seem quite nonspecific. This article reviews pertinent issues concerning diagnosis and management of pediatric cardiac disorders including supraventricular tachycardia, congenital heart disease, prolonged QT syndrome, pericarditis, myocarditis, rheumatic fever, and endocarditis. It also reviews abnormal findings in pediatric electrocardiograms. PMID- 14751482 TI - Heterotopic gestation: another possibility for the emergency bedside ultrasonographer to consider. AB - Emergency bedside ultrasonography has become an important tool in differentiating an intrauterine from an ectopic pregnancy. As the odds of a heterotopic pregnancy were thought to be minute, some ultrasonographers and Emergency Physicians certified in bedside ultrasonography have taught that an intrauterine gestation with fetal heart tones clinches the diagnosis of intrauterine pregnancy. However, with the current rise in heterotopic gestations, this standard of care should be re-evaluated. If emergency bedside ultrasonography reveals an intrauterine gestation in the presence of either a moderate amount of free fluid in the pelvis or a co-existent ovarian cyst, one must consider and search for a heterotopic pregnancy. PMID- 14751483 TI - EMLA-induced methemoglobinemia and systemic topical anesthetic toxicity. AB - This case report illustrates an adult presenting with the simultaneous occurrence of both methemoglobinemia (MetHb) and systemic toxicity from the topical application of local anesthetics while undergoing laser epilation therapy of the legs. The concurrent development of both is considered uncommon in this setting and may have been related to several factors, including her recent previous treatment, increased absorption secondary to abraded skin with the addition of occlusive dressing, and possible alteration of protein binding and drug metabolism due to the use of medications. The clinical manifestations and mechanisms of MetHb and systemic local anesthetic toxicity are discussed. PMID- 14751484 TI - Prevalence of autonomic signs and symptoms in antimuscarinic drug poisonings. AB - Classically described antimuscarinic poisoning signs and symptoms include mydriasis, decreased secretions, ileus, urinary retention, hyperthermia, tachycardia, and altered mental status. These features may be used clinically to assist in the diagnosis of patients with unknown poisonings. We sought to analyze the prevalence of antimuscarinic physical examination findings in evaluating patients presenting with acute poisoning from antimuscarinic agents. We conducted a retrospective, medical record review at two urban tertiary care teaching hospitals. The study population consisted of patients presenting to the Emergency Department with a diagnosis of acute poisoning secondary to medications with known antimuscarinic side effects during a 78-month period between January 1994 and July 2001. Cases were excluded for incomplete medical records or unreliable histories of ingestion, and when concomitant ethanol intoxication was present on laboratory analysis. Clinical information obtained from each patient included vital signs, pupillary size, electrocardiogram abnormalities, the presence of mucous membrane and axillary secretions, initial urine output after bladder catheterization, quality of bowel sounds, mental status changes, the occurrence of seizures and coma, need for orotracheal intubation, and time required for clinical resolution. Diagnostic and therapeutic information including laboratory tests, administration of sodium bicarbonate, and usage of physostigmine was also collected. We identified a total of 345 cases, 213 of which met inclusion criteria. Of these cases, the most common documented findings included decreased secretions in 75.1%, tachycardia in 68.1%, confusion in 49.3%, drowsiness in 48.2%, and hypoactive or absent bowel sounds in 44.6%. Combining signs and symptoms to predict this toxic syndrome was not very reliable. Tachycardia, decreased oral or axillary secretions, and mydriasis proved to be the most predictive trio of clinical signs, but were found in only 28.2% of cases. At least one of these three signs was documented in 94% of our patients. The combination of tachycardia and decreased secretions was the most common pair of findings, recorded in 55.4% of cases. We conclude that the clinical presentation of antimuscarinic syndrome is variable. PMID- 14751485 TI - Electrocardiographic manifestations: diagnosis of atrioventricular block in the Emergency Department. AB - Patients with bradycardia are commonly encountered by the Emergency Physician. Of the possible bradydysrhythmias, atrioventricular blocks (AVB) represent a significant portion of these presentations. In this article, we provide four illustrative cases of patients presenting to the Emergency Department (ED) with AVB. We review the various types of AV block dysrhythmias (1st, 2nd, and 3rd degrees) and their underlying etiologies. This discussion also focuses on the presentation, clinical considerations, management and acute treatment of AVB dysrhythmias in the emergent setting. PMID- 14751486 TI - Cervical epidural abscess associated with massively elevated erythrocyte sedimentation rate. AB - We present a case of an elderly woman who presented with neck pain, low-grade fever, bandemia, and a massively elevated erythrocyte sedimentation rate (ESR) who had a cervical epidural abscess. We believe that the selective use of ESR assisted in narrowing the differential diagnosis, as the patient had no neurological deficits and no predisposing factors such as distal infection, immunosuppression, trauma, or recent surgery. Furthermore, in the literature, an elevated ESR is consistently found in patients with epidural abscesses, whereas clinical findings such as fever, leukocytosis, and neurological deficits are only variably present. PMID- 14751487 TI - Fever. PMID- 14751488 TI - Superior mesenteric vein thrombosis following uncomplicated appendectomy. PMID- 14751489 TI - Twiddler syndrome. PMID- 14751490 TI - The use of brief alcohol interventions in the ED employed by emergency physicians and staff with trauma patients: a suggestion that is a bit naive. PMID- 14751491 TI - "A national epidemic of unassigned patients". PMID- 14751493 TI - A girl with a giant bladder stone. PMID- 14751494 TI - Clinical trials in an emergency setting: implications from the fifth version of the Declaration of Helsinki. AB - Everybody agrees that research is crucial to improve the quality of emergency care. Consent of human subjects for participation in research requires that they fully understand their role and risk, not be coerced, and be allowed to withdraw at any time without penalty. In an emergency situation, informed consent is not always possible but the need for good research data is very high. Here is the ethical difficulty, and a real conflict of values: a population that might ultimately benefit from research cannot consent to the research and are thus excluded from the potential therapeutical advances. Patients at high risk of morbidity or death, with cardiac arrest, shock, head injury, or altered mental status, are evidently incapable of providing an adequate consent, but nevertheless are often in the greatest need of innovative therapy and might be willing to assume some risk for potential benefit. In an attempt to resolve this dilemma, the new version of the Declaration of Helsinki presents updated requirements for the waiver of informed consent and the protection of human subjects in emergency research. PMID- 14751497 TI - Advances in translational radiation oncology: from molecular signaling to cancer cure. PMID- 14751498 TI - International Conference on Translational Research ICTR 2003 Conference Summary: marshalling resources in a complex time. AB - The knowledge, tools, and environment for the practice of radiation oncology are changing rapidly. The National Cancer Institute has articulated the need for a balanced portfolio, including the interrelated components of discovery, development, and delivery. Underpinning practice is the emerging knowledge from molecular, cellular, and tumor biology that is the engine of discovery. The use of high-throughput technologies to analyze biochemical and molecular profiles will ultimately enable the individualization of cancer treatment requiring the appropriate integration of radiation with a range of systemic therapies, including chemotherapy, biologic therapy, and immunotherapy. Technological advances in treatment delivery using photons, brachytherapy, particle therapy, radioisotopes, and other forms of energy require an improved ability to localize the tumor and critical subregions and to ensure necessary tissue immobilization and/or real-time target adjustment. Functional imaging is helping to define tumor characteristics and response to treatment. The development of appropriate radiation oncology treatment requires a wide range of expertise, a multimodality approach, and multi-institutional collaboration to provide improved and cost effective outcome. The delivery of appropriate cancer care to those who need it requires biology and technology but also reaching the underserved populations worldwide. ICTR 2003 demonstrated substantial progress in translational radiation oncology. Faced with financial constraints for research and patient care, the broad field of radiation oncology must continually examine and balance its research and development portfolio and invest in its future leaders to enable it be an important contributor to the future of cancer care. PMID- 14751499 TI - High-tech in radiation oncology: should there be a ceiling? AB - PURPOSE: To analyze some of the limitations to improvement of the outcome of radiotherapy (RT) expected from the introduction of sophisticated treatment planning and delivery technology. METHODS AND MATERIALS: Several recent examples from the literature were analyzed in some detail. Mathematical modeling techniques were used to assess the likely clinical impact of new technologies or biologic principles. The findings of recent randomized controlled trials of RT for prostate, breast, and rectal cancer were analyzed from the perspective of cost-effectiveness and therapeutic gain. RESULTS: The main findings of the analyses may be summarized as follows. Dosimetric precision should aim for a <2% patient-to-patient variability in the delivered dose. Imprecision in clinical target volume definition remains an obstacle for high-precision RT. Functional imaging and novel biologic assays may facilitate a move from a clinical target volume to the real target volume. Improved target volume coverage is mainly important if RT has high effectiveness. Radiation oncology is increasingly becoming evidence based. However, there is still a long way to go. Hypofractionation in adjuvant RT for breast cancer may represent a favorable balance between cost and benefit. Treatment complications are potentially associated with both suffering and high cost. The identification of high-risk patients would improve the cost-effectiveness of high-tech RT aimed at avoiding complications. Conformal RT may allow the introduction of hypofractionation, which, again, could potentially save resources. With improvement in surgery and more screening-detected cancer cases, the number needed to treat increases, and this will directly affect the cost-effectiveness of high-tech RT unless efficient patient selection can be developed. CONCLUSION: Sustained technological refinement is only likely to be cost-effective if the clinical and biologic understanding of patient-to-patient variability in the risk of specific types of failure and the optimal multimodality approach to handle these risks is developed at the same time. Mathematical modeling together with methods from health technology assessment and health economics are useful complements to standard methods from evidence-based medicine. Progress in functional imaging and in basic and clinical cancer biology is likely to provide the tools required for individualized risk-adapted RT. PMID- 14751500 TI - Phosphorylation of histone H2AX as a measure of radiosensitivity. AB - PURPOSE: Phosphorylation of histone H2AX (gammaH2AX) occurs rapidly in response to the presence of DNA double-strand breaks and is thought to recruit repair enzymes to these sites. We examined the possibility that expression of phosphorylated H2AX could provide information on tumor and/or normal tissue sensitivity to radiation. METHODS: Flow cytometry of gammaH2AX antibody-stained single cells was used to measure gammaH2AX intensity in cultured cell lines, tumor cells, and normal tissues. RESULTS: The rate of disappearance of gammaH2AX during the first few hours after irradiation was generally faster in more radioresistant tumor and normal cell lines, but slower in radiosensitive cells and normal tissues from C3H mice. An exception was testis, which showed a high background and rapid loss rate. Levels of gammaH2AX were at least three times higher in well-oxygenated cells than in anoxic cells, and the oxygen concentration that produced a half-maximal response was 0.55%. Hypoxic cells could be detected in SiHa xenografts as a subpopulation with lower expression of gammaH2AX. CONCLUSIONS: Analysis of gammaH2AX has the potential to provide useful information on tumor and normal cell response to ionizing radiation after exposure to clinically relevant doses of radiation. PMID- 14751501 TI - Antisense MDM2 sensitizes prostate cancer cells to androgen deprivation, radiation, and the combination. AB - PURPOSE: Antisense MDM2 (AS) sensitizes a variety of tumor cell types, including prostate cancer, to radiation and chemotherapy. We have previously described that AS enhances the apoptotic response to androgen deprivation (AD) and that this translates into a reduction in overall cell survival, as measured by clonogenic assay. Because AD + radiation (RT) is a key strategy for the treatment of men with high-risk prostate cancer, AS was tested for the ability to sensitize cells to the combination of AD+RT. METHODS AND MATERIALS: LNCaP cells were cultured in vitro in either complete, androgen deprived (AD), or AD+R1881 (synthetic androgen) medium for 2-3 days before AS was administered. Radiation at 5 Gy was given 18-24 h later. Processing of the cells after RT was done at 3 h for Western blots, 24 and 48 h for trypan blue dye exclusion, 18 h for Annexin V staining by flow cytometric analysis, 18 h for Caspase 3+7 quantification by fluorometric assay, and immediately for clonogenic survival measured 12-14 days later. There were 18 treatment groups that were studied: lipofectin control, AS, antisense mismatch (ASM), AD, AD+R1881, and RT in all possible combinations. Statistical comparisons between groups were accomplished with one-way analysis of variance using the Bonferroni test, considering all 18 groups. RESULTS: AS caused a reduction in MDM2 expression and an increase in p53 and p21 expression. Early cell death by trypan blue was found to be reflective of the apoptotic results by Annexin V and Caspase 3+7. AS caused a significant increase in apoptosis over the lipofectin control, AD, and RT controls. Apoptosis was further increased significantly by the addition of AD or RT to AS. When AS, AD, and RT were combined, there was a consistent increase in early cell death over AS+AD and AS+RT by all of the assay methods, although this increase was not significant. Overall cell death measured by clonogenic assay revealed synergistic cell killing of AS+RT beyond that of ASM+RT and RT alone, and AS+RT+AD beyond that of AS+RT, AS+RT+AD+R1881, ASM+RT+AD, and ASM+RT+AD+R1881. CONCLUSION: AS sensitizes cells to AD, RT, and AD+RT and shows promise in the treatment of the full range of patients with prostate cancer. AS has the potential to sensitize the primary tumor to AD+RT and metastasis to AD. PMID- 14751502 TI - Effects of the EGFR/HER2 kinase inhibitor GW572016 on EGFR- and HER2 overexpressing breast cancer cell line proliferation, radiosensitization, and resistance. AB - PURPOSE: Two members of the epidermal growth factor receptor family, EGFR and HER2, have been implicated in radioresistance in breast cancer and other malignancies. To gauge the potential clinical utility of targeting both EGFR and HER2 to control growth and radiosensitize human breast cancers, we examined the effect of a dual EGFR/HER2 inhibitor, GW572016, on the proliferation and radiation response of either EGFR- or HER2-overexpressing human breast cancer cell lines. METHODS AND MATERIALS: Primary human breast cancer cell lines that endogenously overexpress EGFR or HER2 and luminal mammary epithelial H16N2 cells stably transfected with HER2 were evaluated for the effect of GW572016 on inhibition of ligand-induced or constitutive receptor phosphorylation, proliferation, radiosensitization, and inhibition of downstream signaling. RESULTS: GW572016 inhibited constitutive and/or ligand-induced EGFR or HER2 tyrosine phosphorylation of all five cell lines, which correlated with the antiproliferative response in all but one cell line. GW572016 radiosensitized EGFR-overexpressing cell lines, but HER2-overexpressing cells were unable to form colonies after brief exposure to GW572016 even in the absence of radiation, and thus could not be evaluated for radiosensitization. One cell line was resistant to the antiproliferative and radiosensitizing effects of GW572016, despite receptor inhibition. Exploration of potential mechanisms of resistance in SUM185 cells revealed failure of GW572016 to inhibit downstream ERK and Akt activation, despite inhibition of HER2 phosphorylation. In contrast, sensitive HER2 overexpressing cell lines demonstrated inhibition of both ERK and Akt phosphorylation. CONCLUSION: GW572016 potently inhibits receptor phosphorylation in either EGFR- or HER2-overexpressing cell lines and has both antiproliferative and radiosensitizing effects. Resistance to GW572016 was not due to a lack of receptor inhibition, but rather with a lack of inhibition of ERK and Akt, suggesting that measurement of inhibition of crucial signaling pathways may better predict response than inhibition of receptor phosphorylation. The SUM185 cell line provides a valuable model for studying mechanisms of resistance of EGFR/HER2 inhibitor therapy. PMID- 14751503 TI - Radiosensitization of vinorelbine and gemcitabine in NCI-H460 non-small-cell lung cancer cells. AB - PURPOSE: Results from recent clinical studies of gemcitabine and vinorelbine have encouraged the use of this combination concurrently with radiotherapy in the treatment of non-small-cell lung cancer, although preclinical data are limited. The present study aimed to quantify the in vitro interaction and radiosensitizing effect of gemcitabine and vinorelbine individually and in combination. METHODS AND MATERIALS: Cytotoxicity was measured by exposing NCI-H460 cells to gemcitabine and/or vinorelbine simultaneously or sequentially, followed by irradiation at 0-10 Gy. Clonogenic cell survival assays were performed. Flow cytometry was used to measure the effects of both drug and radiation on cell cycle distribution. Apoptosis was assessed by morphologic criteria, by sub-G1 changes using flow cytometry assay, and by Annexin-V binding assay. RESULTS: Both drugs showed single-agent activity against NCI-H460 cells and targeted different phases of the cell cycle. When both drugs were used in combination, they showed schedule-dependent interaction. An antagonistic effect was observed with simultaneous exposure to the two drugs. The optimum combination schedule was sequential exposure to vinorelbine followed by gemcitabine 24 h later. Both drugs showed radiosensitization effects. The radiosensitization effect of gemcitabine was evident when radiation was given immediately after 4-h incubation. However, the radiosensitization effect of vinorelbine was time dependent and observed with radiation given at 24 h postincubation. Apoptosis induced by gemcitabine increased gradually, reaching 20% at 72 h posttreatment. In contrast, apoptotic cell death was an early feature in vinorelbine-treated cells, reaching approximately 40% at 24 h. CONCLUSIONS: The individual cytotoxic effects of gemcitabine and vinorelbine on NCI-H460 cells are phase specific, and the combined effect of gemcitabine and vinorelbine is sequence dependent. The radiosensitizing effects of both drugs seem to be related to enhanced apoptosis. PMID- 14751504 TI - Novel radiosensitizers for locally advanced epithelial tumors: inhibition of the PI3K/Akt survival pathway in tumor cells and in tumor-associated endothelial cells as a novel treatment strategy? AB - In locally advanced epithelial malignancies, local control can be achieved with high doses of radiotherapy (RT). Concurrent chemoradiotherapy can improve tumor control in selected solid epithelial adult tumors; however, treatment-related toxicity is of major concern and the therapeutic window often small. Therefore, novel pharmacologic radiosensitizers with a tumor-specific molecular target and a broad therapeutic window are attractive. Because of clonal heterogeneity and the high mutation rate of these tumors, combined treatment with single molecular target radiosensitizers and RT are unlikely to improve sustained local tumor control substantially. Therefore, radiosensitizers modulating entire tumor cell survival pathways in epithelial tumors are of potential clinical use. We discuss the preclinical efficacy and the mechanism of three different, potential radiosensitizers targeting the PTEN/PI3K/Akt survival pathway. These compounds were initially thought to act as single-target agents against growth factor receptors (PKI 166 and PTK 787) or protein kinase C isoforms (PKC 412). We describe an additional target for these compounds. PKI 166 (an epidermal growth factor [EGF] receptor inhibitor) and PKC 412, target the PTEN/PI3K/Akt pathway mainly in tumor cells, and PTK 787 (a vascular endothelial growth factor [VEGF] receptor inhibitor) in endothelial cells. Even for these broader range molecular radiosensitizers, the benefit could be restricted to human epithelial tumor cell clones with a distinct molecular profile. Therefore, these potential radiosensitizers have to be carefully tested in specific model systems before introduction in early clinical trials. PMID- 14751506 TI - The bioreductive agent RH1 and gamma-irradiation both cause G2/M cell cycle phase arrest and polyploidy in a p53-mutated human breast cancer cell line. AB - PURPOSE: RH1 is a newly developed bioreductive agent, and its bioactivation is mediated by the enzyme DT-diaphorase (DTD). We have shown previously that RH1 is highly cytotoxic against cells expressing high DTD, using the p53-mutated MDA231 human breast cancer cell line transfected with the DTD gene (D7 cells). We now report that both RH1 and gamma-irradiation cause D7 cells to arrest in the G2/M cell cycle phase and undergo polyploidy. The latter is a way of p53-mutated cells responding to DNA-damaging agents. Only a small proportion of the polyploid cells are clonogenic, hence polyploidy may contribute to the reproductive failure of the cells after RH1 and irradiation. Thus, we investigated the effect of RH1 and gamma-irradiation on the formation of polyploid cells and a sub-G1 population (as a measure of apoptosis) in relation to the G2/M cell cycle block. METHODS AND MATERIALS: MDA231 D7 cells were treated using a range of RH1 doses. The cells were irradiated using 2 Gy or 5 Gy gamma-rays either as a single dose or in combination with RH1. An IC(90) dose (dose to kill 90% of the cells) of RH1 was administered for 3 h followed by irradiation after a further 24 h. Subsequent changes in cell cycle and polyploidy (DNA content in excess of that of G2/M cells) were examined. RESULTS: Treatment of D7 cells with the RH1 resulted in 60 70% of cells arrested in the G2/M phase of the cell cycle by 24 h, which decreased to control levels by 48 h. Irradiation with 2 Gy and 5 Gy caused a similar G2/M block at 12-24 h, which was followed by a sharp decline at 24-48 h. In contrast, the same dose of radiation combined with RH1 held the cells in the G2/M phase up to 48 h, and this pattern reached pretreatment levels at 72-96 h. Most control cells were found to contain a small number of spontaneously arising polyploid cells. The development of polyploid cells was evident from 12 h after all treatments and showed a significant increase at 48 h and subsequently. As opposed to this, apoptosis measured by the sub-G1 cell population in DNA analyses showed a tendency to increase according to the elapsed time for each group of treatments. Single treatments with RH1 caused a significant increase in the apoptotic population between 48 and 120 h. The first significant increase in apoptosis was observed at 48 h for 5 Gy, 2 Gy + RH1, and 5 Gy + RH1 treatments, and showed a tendency to increase further at later times, but the 2 Gy dose gave an earlier apoptotic peak at 24 h, which decreased to 96 h. The addition of RH1 to the irradiation did not increase the formation of polyploid cells or apoptosis compared with radiation alone (2 Gy vs. RH1 + 2 Gy or 5 Gy vs. RH1 + 5 Gy). The higher dose of irradiation (5 Gy vs. 2 Gy) resulted in a significantly higher proportion of polyploid cells (but not of apoptotic cells) when used alone or in combination (5 Gy + RH1 vs. 2 Gy + RH1). CONCLUSIONS: Both RH1 and gamma irradiation, individually and in combination, showed a significant G2/M block in MDA231 D7 breast cancer cells. The formation of polyploid cells was dependent more on the radiation dose rather than on the pretreatment with RH1. The polyploid cell population was observed after the G2/M cell cycle phase arrest, and it preceded the late increase of the apoptotic cell population. The role of polyploidy in cell reproductive failure in the total cell population is not known, but it appears to contribute to cytotoxicity in cells released from the G2/M cell cycle phase block. PMID- 14751505 TI - Potentiation of tumor response to radiation or chemoradiation by selective cyclooxygenase-2 enzyme inhibitors. AB - Cyclooxygenase-2 (COX-2) is an enzyme expressed primarily in pathologic states, such as inflammatory disorders and cancer, where it mediates prostaglandin production. Its overexpression is associated with more aggressive biologic tumor behavior and adverse patient outcome. Increasing evidence shows that agents that selectively inhibit COX-2 enhance tumor response to radiation or chemotherapeutic agents. This article gives an overview of some of this evidence. In addition, we describe new results showing that celecoxib, a selective COX-2 inhibitor, enhanced response of A431 human tumor xenografts in nude mice to radiation by an enhancement factor (EF) of 1.43 and to the chemotherapeutic agent docetaxel by an EF of 2.07. Celecoxib also enhanced tumor response when added to the combined docetaxel plus radiation treatment (EF = 2.13). Further experiments showed that selective COX-2 inhibitors enhanced tumor cell sensitivity to ionizing radiation, involving inhibition of cellular repair from radiation damage and cell cycle redistribution as mechanisms for some cell types. The results show that selective COX-2 inhibitors have the potential to improve tumor radiotherapy or radiochemotherapy, and this therapeutic strategy is currently under clinical testing. PMID- 14751507 TI - Influence of hypoxia on TRAIL-induced apoptosis in tumor cells. AB - PURPOSE: Tumor hypoxia reduces the efficacy of radiotherapy, many types of chemotherapy, and tumor necrosis factor-alpha (TNF-alpha). TRAIL (TNF-alpha related apoptosis-inducing ligand) is a ligand for death receptors of the TNF superfamily shown to be selectively toxic for tumor cells and thereby a promising antineoplastic tool. The impact of hypoxia on TRAIL-induced apoptosis was examined in this study. METHODS AND MATERIALS: Apoptosis induction and growth rates of various tumor cell lines under hypoxia were evaluated in vitro. Biologically effective induction of hypoxia was verified by determination of hypoxia-inducible factor-1 (HIF-1) activation. The efficacy of TRAIL- and radiation-induced apoptosis under different oxygen conditions was quantified in vitro. The impact of Bcl-2 on TRAIL-induced apoptosis under hypoxia or normoxia was evaluated by comparing cells expressing Bcl-2 with a vector control. RESULTS: Moderate hypoxia caused no growth retardation or apoptosis, but led to activation of HIF-1 as a prerequisite of hypoxic gene induction. Cellular responses to TRAIL differed considerably among the cell lines tested. Hypoxia reduced radiation induced, but not TRAIL-induced, apoptosis in the tested cell lines. Hypoxia did not induce Bcl-2 expression. Bcl-2 had a minor impact on the efficacy of TRAIL induced apoptosis. CONCLUSION: Taken together, the data indicate that TRAIL is clearly effective under conditions of proven hypoxia. PMID- 14751508 TI - In vitro and in vivo evaluation of novel antitumor prodrugs of 5-fluoro-2' deoxyuridine activated by hypoxic irradiation. AB - PURPOSE: We previously developed a novel antitumor prodrug that has a 2-oxopropyl substituent at the N(1) position of 5-fluorouracil (5-FU) and releases 5-FU via one-electron reduction on hypoxic irradiation. Although the compound was effective in vivo, its activity against murine tumors was not high enough to warrant clinical studies. Therefore, we developed a similar family of radiation activated prodrugs of 5-fluoro-2'-deoxyuridine (FdUrd), which is generally more potent than 5-FU, and investigated their radiation chemical reactivity and in vitro and in vivo effects. METHODS AND MATERIALS: Compounds bearing various 2 oxoalkyl substituents at the N(3) position of FdUrd were synthesized and investigated. After aerobic or hypoxic irradiation to the prodrugs dissolved in water or culture medium, release of FdUrd was measured using high-performance liquid chromatography. To investigate in vitro cytotoxicity, SCCVII and EMT6 cells in culture were irradiated in the presence of the prodrug under aerobic or hypoxic conditions, and then kept with the compound for 24 h. Cell survival was then measured using a colony assay. To investigate in vivo effects, the drug was injected intraperitoneally at a dose of 100 or 300 mg/kg into Balb/c mice bearing EMT6 tumors 30 min before irradiation. The tumor growth delay-time was then assessed. RESULTS: In vitro, the prodrugs released FdUrd at G-values (molar numbers of molecules produced by 1 J of radiation energy) of 1.6-2.0 x 10(-7) mol/J after hypoxic irradiation. The G-values for FdUrd release with hypoxic irradiation were about 100-fold greater than those with aerobic irradiation. Among the prodrugs tested, OFU106 bearing a 2-oxocyclopentyl substituent released the highest amount of FdUrd in the culture medium, and it was subjected to further in vitro and in vivo assays. Although OFU106 administered alone showed no cytotoxicity up to a concentration of 0.2 mM, it produced an enhanced cytotoxic effect when administered before hypoxic irradiation and kept with the cells for 24 h. The enhancement ratios calculated at the surviving fraction of 1% were 1.35 1.4 at 0.04 mM and 1.45-1.5 at 0.2 mM. In vivo, however, administration of OFU106 (100 or 300 mg/kg) before 20 Gy of irradiation did not produce marked growth delays compared with 20 Gy of radiation alone. CONCLUSION: On hypoxic irradiation in vitro, the prodrugs of FdUrd were activated as efficiently as were the prodrugs of 5-FU, but marked in vivo effects could not be detected. This strategy of prodrug design should be used in further development of radiation-activated prodrugs of more potent anticancer agents. PMID- 14751509 TI - Tumor-line specific pO(2) fluctuations in human melanoma xenografts. AB - PURPOSE: A large number of studies have demonstrated that tumors are heterogeneous in oxygen tension (pO(2)) and may develop regions with chronically or acutely hypoxic cells during growth. In the present study, it was investigated whether experimental tumors of different lines may show characteristic pO(2) fluctuation patterns and hence may differ with respect to the kinetics of acute hypoxia. METHODS AND MATERIALS: A total of 70 xenografted tumors of two human melanoma lines (A-07 and R-18) were included in the study. Tissue pO(2) was measured simultaneously in two positions in each tumor for periods of at least 60 min using a two-channel fiberoptic oxygen-sensing device (OxyLite 2000, Oxford Optronix, Oxford, UK). RESULTS: The mean pO(2) was calculated for each pO(2) trace, and this parameter was significantly greater in A-07 than in R-18 tumors (p <0.000001). Fluctuations in pO(2) around 3, 5, or 10 mm Hg were seen in a large fraction of the tumors of both lines. The pO(2) fluctuation frequency differed among individual traces from 0 to 20/h (A-07) and from 0 to 12/h (R-18) and was significantly greater in A-07 than in R-18 tumors (p = 0.0026). The absolute amplitude of the pO(2) fluctuations ranged from 1 to 16 mm Hg (A-07) and 1 to 33 mm Hg (R-18) and did not differ between the tumor lines. The relative amplitude was significantly higher in R-18 than in A-07 tumors (p <0.000001). The pO(2) values recorded simultaneously in the same tumor were in general not temporally coordinated. CONCLUSION: Experimental tumors of different lines may show individual and characteristic pO(2) fluctuation patterns. The pO(2) fluctuations may result in regions with acutely hypoxic cells. The kinetics of the acute hypoxia may differ among tumors of different lines, individual tumors of the same line, and different regions within the same tumor. PMID- 14751510 TI - PARP-1, PARP-2, and the cellular response to low doses of ionizing radiation. AB - PURPOSE: Poly(ADP-ribose) polymerase-1 (PARP-1) is rapidly and directly activated by single-strand breaks and is required for efficient base excision repair. These properties indicate that inhibition of PARP-1 might enhance the cellular response to low doses of radiation. We tested the effect of chemical inhibition of PARP-1 on low-dose clonogenic survival in a number of cell lines and the low-dose radiation response of a PARP-1 knockout murine cell line. METHODS AND MATERIALS: Clonogenic cell survival of V79-379A and CHO-K1 hamster fibroblasts, T98G and U373-MG human glioma cells, and 3T3 mouse embryo fibroblast PARP-1 knockout cells was measured using a precise flow cytometry-based plating assay. Chemical inhibitors of PARP enzymes were tested for their effect on clonogenic survival after a range of ionizing radiation doses. RESULTS: Chemical inhibition of PARP activity induced marked radiosensitization of V79, CHO, and exponentially growing T98G cells in the 0.05-0.3-Gy range. This effect was not seen in U373 cells or in confluent T98G populations. Low-dose radiosensitization was not apparent in PARP 1 knockout cells. CONCLUSION: Low-dose radiosensitization of actively dividing tumor cells by PARP-1 inhibitors suggests that they may have a role in enhancing the efficacy of ultrafractionated or low-dose-rate radiotherapy regimens. We hypothesize that PARP-2 compensates for the absence of PARP-1 in the knockout cell line. PMID- 14751511 TI - Microarray analysis to identify molecular mechanisms of radiation-induced microvascular damage in normal tissues. AB - PURPOSE: Radiation-induced vascular injury can be a serious problem for cancer survivors. In capillary vessels, this manifests as telangiectasia, causing cosmetic problems when occurring in the skin and more serious problems, e.g. excessive bleeding requiring surgery, when occurring in rectal or bladder mucosa. In addition, thrombotic, inflammatory, and fibrogenic events play an important role in the development of late radiation injury in many tissues. However, the sequence of these events and the relationship between various mechanistic pathways is unclear. The purpose of this project is to identify genes that are differentially expressed in tissues with manifest vascular damage, with the ultimate goal of intervening in this process to block the progressive development of tissue injury. METHODS AND MATERIALS: Microarray experiments were performed using amplified RNA isolated from irradiated mouse kidney and rectum, and from sham-irradiated controls, at 10 and 20 weeks after treatment. Tissue samples were also taken for histologic evaluation of vascular damage at 10, 20, and 30 weeks after irradiation. Expression profiles for irradiated and sham-irradiated samples were compared, and differentially expressed genes were identified after normalization procedures, using information from straight color, color reverse, and self-self experiments. The extent of overlap in expression profiles for kidney and rectum during the phase of vascular damage was also examined. RESULTS: The mouse kidney experiments showed upregulation of 31 genes at 10 weeks and 42 upregulated genes at 20 weeks. Only 20 genes showed significantly increased expression at both time points. Some of these genes were of particular interest in terms of their known involvement in vascular injury and signal transduction pathways. Irradiated mouse rectum had 278 upregulated genes at 10 weeks and 86 upregulated genes at 20 weeks. Only 19 of the genes upregulated during the period of identified telangiectasia (10-20 weeks) were common to both tissues. These included jagged 1 and Kruppel-like factor 5 (KLF5), which are reported to play a role in vascular development and remodeling. CONCLUSIONS: Microarray analysis of RNA from irradiated normal tissues is an effective tool for identifying new genes of potential interest in the development of late tissue injury. Such experiments should be regarded as generating testable hypotheses for mechanisms of radiation induced injury. PMID- 14751512 TI - Acute induction of inflammatory cytokine expression after gamma-irradiation in the rat: effect of an NF-kappaB inhibitor. AB - PURPOSE: The pathologic changes within the intestinal muscle layer may be at the origin of the cytokines that account for acute radiation-induced inflammation. We were specifically interested in evaluating the efficacy of an inhibitor of nuclear transcription factor kappa B (NF-kappaB) activation that is involved in regulating cytokine expression. METHODS AND MATERIALS: Cytokine expression was analyzed in the ileal muscularis layer by reverse transcriptase-polymerase chain reaction (RT-PCR) at 3 h, 6 h, and 3 days after a 10-Gy gamma whole-body irradiation of rats. Caffeic acid phenethyl ester (CAPE) was injected intraperitoneally (30 mg/kg) 15 min before irradiation and once a day for 3 days. RESULTS: Interleukin (IL)-1beta, tumor necrosis factor alpha (TNF-alpha), and IL 6 mRNA increased at 3 h and 6 h after irradiation, and expression of IL-6 and IL 8 was elevated at 3 days. On the other hand, levels of the anti-inflammatory cytokine IL-10 were markedly lower on Day 3. Overexpression of IL-6 on Day 3 was combined with upregulation of the IL-6 receptors (gp130/gp80) and suppressor of cytokine signaling-3 (SOCS3) genes. CAPE treatment did not significantly change IL-1beta or TNF-alpha expressions in the irradiated rats; it increased IL-10 expression at 6 h but had no effect on it on Day 3. CAPE treatment inhibited the radiation-induced expression of IL-6, IL-6 receptors (IL-6rs), and SOCS3 at 3 days. CONCLUSION: In vivo, irradiation induced a cascade of inflammatory responses that involved the transcription factor NF-kappaB; this inflammation was reduced by CAPE treatment. PMID- 14751514 TI - For X-irradiated normal human fibroblasts, only half of cell inactivation results from chromosomal damage. AB - PURPOSE: To study the relationship between residual double-strand breaks (dsbs), chromosomal damage, and cell inactivation for X-irradiated normal human fibroblasts. METHODS AND MATERIALS: The experiments were performed with 12 normal human fibroblast strains and, for comparison, a fibroblast line from a LiFraumeni patient (LFS2800), a squamous cell carcinoma line (FaDu), and CHO cells. Cells were irradiated in plateau phase, which was followed by immediate or delayed (14 h) plating. Chromosomal damage was measured by metaphase technique and loss of proliferative capacity by colony-forming assay. The data obtained were compared with residual double-strand breaks measured previously (Dikomey et al. IJROBP 2000;46:481-490). RESULTS: For each fibroblast strain, the number of lethal chromosome aberrations (CAs) increased with dose, but with a substantial variation among the strains (coefficient of variation = 20%-26%). The number of lethal aberrations was significantly correlated with the number of residual dsbs measured for the same strain (r(2) = 0.71, p = 0.0006). The residual dsbs were assumed to represent both non- and also mis-rejoined dsbs. There was a significant correlation between lethal aberrations and cell survival, but only for delayed and not immediate plating (r(2) = 0.69, p < 0.0008 vs. r(2) = 0.19, p = 0.16). For delayed plating, the ratio between lethal events (LEs) and CAs amounted to LE:CA = 2.0 +/- 0.05:1, indicating that on average, only half of cell inactivation resulted from chromosomal damage. The other 50% was attributed to the p53-dependent permanent G1 arrest, because cells lacking in functional p53 (LFS2800, FaDu, CHO) showed a ratio of LE:CA = 1.01 +/- 0.02:1. CONCLUSION: On average, up to 50% of the inactivation of X-irradiated normal human fibroblasts is a result of lethal chromosome aberrations, whereas the rest is due to a p53 dependent process, probably permanent G1 arrest. PMID- 14751513 TI - Cellular kinetics of murine lung: model system to determine basis for radioprotection with keratinocyte growth factor. AB - PURPOSE: Normal tissue toxicity remains a dose limitation for cancer radiotherapy and chemoradiotherapy. Growth factors offer a novel means of mitigating normal tissue radiotoxicity. In particular, keratinocyte growth factor (rHuKGF), whose proliferative activity is restricted to epithelial cells, holds promise on the basis of the findings of preclinical models of epithelial cytoprotection and the clinical developments to date. We report the radioprotection of murine lung by an increase in tissue cellularity after rHuKGF-induced proliferation. METHODS AND MATERIALS: Flow cytometric and image analysis techniques after bromodeoxyuridine labeling were used to estimate proliferative parameters. Our specialized analytical methods measure not only labeling indexes, but also the durations of S and G(2)+M phases, potential doubling times, and the net cell production rate. Image analysis techniques were used to identify the specific cell types that were proliferating (type II pneumocytes). RESULTS: Lung labeling index control values (0.5%) rose to a maximum (5.5%) at 3 days after intratracheal rHuKGF, returning to normal by Day 7. The potential doubling time fell from 66 days to 4.4 days. The net cell production rate rose from a control value of 1%/d to >15%/d by Day 3. This resulted in a nearly twofold increase in alveolar epithelial cellularity, which remained significantly elevated on Day 7. Saline-treated control animals exhibited no significant changes in the proliferative parameter values or cellularity. On the basis of these data, mice were irradiated, solely to the thorax, with ranges of single doses of 250 kVp X-rays 7 days after either intratracheal administration of 5 mg/kg rHuKGF or phospate-buffered saline. This interval was chosen because the proliferative response of the type II cells was finished but the cellularity of the lung remained increased. Pretreatment with rHuKGF extended the latent period before onset of pneumonitis after all radiation doses. rHuKGF treatment 7 days before thoracic irradiation significantly protected against pneumonitis (median effective dose 13.7 Gy, 95% confidence limit 13.4-14.0) compared with the control pretreatment with phosphate-buffered saline (median effective dose 12.8 Gy, 95% confidence limit 12.6-13.1). CONCLUSION: The data showed that an increase in tissue cellularity, caused by rHuKGF treatment before irradiation, protected the lung from damage due to pneumonitis. PMID- 14751515 TI - Arrest of human lung fibroblasts in G2 phase after irradiation is regulated by converging phosphatidylinositol-3 kinase and beta1-integrin signaling in vitro. AB - PURPOSE: Cell-matrix interactions might confer cellular radioresistance in vitro. As a function of radiation, the impact of fibronectin (FN) and phosphatidylinositol-3 kinase (PI3K)-related signaling on survival, the cell cycle, and the beta1-integrin signaling kinases integrin-linked kinase (ILK), protein kinase Balpha/Akt (PKBalpha/Akt), and glycogen synthase kinase-3beta (GSK 3beta) was examined in normal lung fibroblasts in vitro. METHODS AND MATERIALS: Normal human CCD32 lung fibroblasts grown on polystyrene, FN, or poly-L-lysine were irradiated with 0-8 Gy. Colony forming assays, flow cytometric DNA analysis, immunoblotting (Chk1, Chk2, Cdc25C, Cdk1, 14-3-3, p53, p21), and protein kinase assays (ILK, PKBalpha/Akt, GSK-3beta) were performed with or without PI3K inhibition using LY294002 or wortmannin. RESULTS: FN significantly elevated clonogenic survival of CCD32 cells after irradiation compared with polystyrene or poly-L-lysine. FN improved accumulation of irradiated cells in G(2)/M (60%) compared with polystyrene (43%). LY294002 prevented radiation-dependent G(2) blockage on polystyrene; on FN, G(2) arrest was only slightly reduced. Radiation- and PI3K inhibition-related changes in expression and phosphorylation of the various cell cycle proteins tested correlated with the cell cycle data acquired. The kinase activities of ILK, PKBalpha/Akt, and GSK-3beta were strongly induced by irradiation on polystyrene, but not on FN, which was a result of a FN-mediated increase of basal kinase activities. In contrast to polystyrene, FN enabled radiation-dependent induction of ILK and GSK-3beta in a PI3K-independent manner. CONCLUSION: The data indicate a tight convergence of cell-matrix and cell-growth factor interactions that seem to optimize the cellular responsiveness to ionizing radiation in terms of survival and G(2) arrest. ILK, PKBalpha/Akt, and GSK-3beta involved in integrin signaling were uncovered as new molecular factors within the cellular radiation response. Our findings might also provide insight into normal tissue effects and cellular radioresistance. PMID- 14751516 TI - Target cells in radiation pneumopathy. AB - Radiation pneumopathy is the reaction of the organ lung to radiation effects in various target cells. It starts as an exudative inflammation, with the clinical picture of interstitial pneumonia 6-12 weeks after irradiation, and proceeds to a productive chronic inflammation lasting several months and terminating, as other chronic inflammations do, in scar formation, called lung fibrosis. Lung fibrosis is the common end point after lung damage from a wide range of damaging agents. The pathogenetic process leading to the signs and symptoms of radiation pneumopathy is an integrated response of the complex organization of lung tissue. Clinical and pathologic data in patients do not support the hypothesis that the early inflammatory phase of radiation "pneumonitis" and late "radiation fibrosis" are independent pathogenetic processes in the same way as acute radiodermatitis and subcutaneous fibrosis are separate pathologic entities. The target cell population that initiates the pathogenetic process in the lung is not known, and it has been suggested that no single identifiable target exists. The entire process is the result of complex functional alterations in endothelial cells, pneumocytes, macrophages, and other resident and transient cells. No evidence has been found for a role of stem cell sterilization, for impaired transit cell proliferation, or for hypoplasia, which is the hallmark of other acute inflammatory normal tissue damage (i.e., in the mucosa). The radiobiologic concepts developed in cellular radiobiology are not adequate for the quantitative analysis of radiation pneumopathy. A new analytical framework based on structurally defined intercellular interaction by signaling molecules and their activation needs to be developed. This would not be only an abstract radiobiologic paradigm but would be the key to the development of potential therapeutic interventions in irradiated patients. PMID- 14751517 TI - Primary fibroblasts from BRCA1 heterozygotes display an abnormal G1/S cell cycle checkpoint following UVA irradiation but show normal levels of micronuclei following oxidative stress or mitomycin C treatment. AB - PURPOSE: There is evidence to suggest that the breast cancer predisposing gene, BRCA1, is involved in cell cycle control and the response to damage but mouse brca1+/- heterozygotes have no distinctive phenotype. Here the response to the three forms of cellular stress was examined in primary human fibroblasts from individuals with a +/+ or +/- genotype for BRCA1. METHODS AND MATERIALS: Fibroblasts from individuals carrying mutations in the BRCA1 gene were compared with those from those wild-type for BRCA1 in their response to long wavelength uv (UVA), hydrogen peroxide, and mitomycin C (MMC). Cell cycle progression and micronucleus formation (MN) were used as end points. RESULTS: After UVA treatment there was no difference between +/- and +/+ cells in the initial fall in DNA synthetic activity (G(1) arrest) but the reentry into S-phase was restored at a faster rate in the BRCA1+/- cells after UVA exposure. Thus, for three normal (+/+) cell lines irradiated in monolayer, S-phase values averaged 15 +/- 3.7% 14 h post-UVA (1 x 10(5) J/m(2)), as compared with 35.7 +/- 1.9 (range) for two BRCA1(+/-) strains. Because a defective G(1)/S checkpoint in BRCA1 heterozygotes could lead to a greater proportion of S-phase cells with unrepaired DNA damage (strand breaks) and a resultant increase in chromosomal instability, the frequency of micronuclei induced by UVA was examined. Three normal (+/+) and three mutant (+/-) strains (two of which were used in the cell cycle experiments) produced mean micronuclei frequencies of 0.077 +/- 0.016 and 0.094 +/- 0.04/binucleate cell respectively (not statistically significant), 48 h after UVA exposure. No differences were found between BRCA1+/+ and +/- cells in MN formation after treatment with MMC or hydrogen peroxide. CONCLUSION: Our data suggest a defective G(1)/S checkpoint in cells from BRCA1 heterozygotes in response to UVA although this is not reflected in genomic instability as measured by micronuclei induction after oxidative stress or MMC treatment. PMID- 14751518 TI - Induction of p53 and p21 proteins by gamma radiation in skin fibroblasts derived from breast cancer patients. AB - PURPOSE: To test the hypothesis that breast cancer patients could have a predisposing defect in the induction of TP53 tumor suppressor protein (known as p53) that could be a marker of genetic instability and increased cancer susceptibility. The accumulation of p53 was triggered by ionizing radiation, and its transcriptional activity was estimated from the induction of CDKN1A (known as p21). METHODS AND MATERIALS: Using Western blot, we studied the radiation induction of p53 and p21 proteins in 28 fibroblast cell strains derived from 7 healthy donors (Control), 3 ataxia telangiectasia (AT), 3 AT heterozygous, and 15 breast cancer patients. Contact-inhibited plateau phase cells were irradiated with 6 Gy, and proteins were assessed 3, 6, and 24 h later. RESULTS: The control group showed a high induction of p53 at 3 h postirradiation followed by a relative decrease at 6 h, and approached the basal level at 24 h. This was associated with a parallel induction in p21. The response of fibroblasts derived from the breast cancer group was not significantly different from the control. Similar results were obtained with the AT heterozygous group at 3 h postirradiation; however, sustainable increase in p53 and p21 induction was observed at 24 h. AT cells showed a statistically significant highly reduced accumulation of p53 and a delayed and reduced induction of p21 (p < or = 0.05). Furthermore, a correlation was observed between these two proteins at the 3-h induction point, confirming the high dependence of p21 on p53. CONCLUSION: Although severe deficiency in p53 and p21 induction was observed in fibroblasts derived from the cancer-prone AT syndrome, these results do not support the assumption that breast cancer patients have a predisposing defective induction in these proteins after ionizing irradiation. PMID- 14751519 TI - Changes in the effect of dose fractionation during daily fractionated irradiation: studies in mouse oral mucosa. AB - PURPOSE: The aim of the present study was to quantify the fractionation effect in mouse oral mucosa during a daily fractionated protocol. METHODS AND MATERIALS: Irradiation of the snout of C3H mice was performed with 5 x 3 Gy/week. In the first experiment, graded test doses were applied to the lower tongue on Days 4, 7, 11, 14, or 18. Subsequently, a split-dose experiment was performed on the same days with 1, 2, 3, or 5 fractions (graded doses). RESULTS: The ED50 (dose expected to induce ulcer in 50% of the mice) for untreated mucosa was 10.7 +/- 1.0 Gy. Fractionated irradiation with 5 x 3 Gy/week yielded an ED50 for top-up irradiation of 6.5 +/- 1.8 Gy, 6.8 +/- 0.9 Gy, 5.3 +/- 2.1 Gy, 7.3 +/- 1.9 Gy, and 7.5 +/- 1.3 Gy on Days 4, 7, 11, 14, and 18. The ED50 values for split-dose irradiation on Day 4 increased from 6.24 +/- 1.48 Gy (1 fraction) to 9.96 +/- 1.47 Gy (5 fractions). Similarly, an increase from 4.04 +/- 1.56 Gy to 10.07 +/- 1.9 Gy was found on Day 11, and from 5.84 +/- 2.37 Gy to 9.81 +/- 2.29 Gy on Day 18. After the weekend breaks, values between 6.22 +/- 1.60 Gy (1 fraction) and 4.7 +/- 1.95 Gy (5 fractions) were observed on Day 7, and between 5.62 +/- 1.36 Gy and 5.98 +/- 2.01 Gy on Day 14. CONCLUSIONS: These results indicate that the fractionation effect in oral mucosa is consistently lost over the weekends and restored during the treatment weeks. PMID- 14751520 TI - Thrombospondin-1 treatment prevents growth of dormant lung micrometastases after surgical resection and curative radiation therapy of the primary tumor in human melanoma xenografts. AB - PURPOSE: Intradermal D-12 human melanoma xenografts develop pulmonary micrometastases in BALB/c nu/nu mice, and these metastases are kept dormant for prolonged times, because the primary tumor secretes thrombospondin-1 (TSP-1) into the blood circulation of the host. In this study, we report on the development of macroscopic metastases after surgical resection and curative radiation treatment of the primary tumor, the mechanisms involved, and the effects of treating the host with exogenous TSP-1 after the eradication of the primary tumor. METHODS AND MATERIALS: Xenografted tumors of the D-12 human melanoma were used as tumor model. Macroscopic metastases were scored by using a stereomicroscope. Micrometastases were detected by histologic examinations. Angiogenesis was studied by using an intradermal angiogenesis assay. Apoptotic endothelial cells were detected by immunohistochemistry by using an in situ apoptosis detection kit. RESULTS: Surgical resection as well as curative radiation treatment of the primary tumor resulted in accelerated growth of dormant micrometastases. This growth could be prevented by treating the host with exogenous TSP-1 after the surgery or the irradiation. Endogenous and exogenous TSP-1 prevented metastatic growth by suppressing angiogenesis, i.e., by inducing apoptosis in activated endothelial cells adjacent to dormant micrometastases. CONCLUSIONS: TSP-1 has antiangiogenic and antimetastatic effects that should be investigated further in clinical studies. Cancer patients with TSP-1-producing primary tumors may benefit from combined local treatment and antiangiogenic/antimetastatic treatment with TSP-1. PMID- 14751521 TI - Target validation of cytochrome P450 CYP1B1 in prostate carcinoma with protein expression in associated hyperplastic and premalignant tissue. AB - PURPOSE: To investigate the localization and distribution of cytochrome P450 CYP1B1 protein expression in patients diagnosed with prostate carcinoma compared to those with bladder carcinoma. To validate CYP1B1 as a molecular target for the development of selective cancer therapeutics for use in combination with radiation. METHODS AND MATERIALS: Prostatectomy specimens (n = 33) of moderate Gleason grade (3 + 3 and 3 + 4) were analyzed immunohistochemically for CYP1B1 protein expression using a specific monoclonal antibody for the enzyme. The intensity of CYP1B1 staining was assessed both semiquantitatively using visual scoring and quantitatively by spectral imaging microscopy using reference spectra and compared with bladder carcinoma (n = 22). RESULTS: CYP1B1 protein expression was present in 75% of prostate carcinomas (n = 27) compared to 100% of bladder carcinomas (n = 22). In both cases, CYP1B1 protein expression was heterogeneous and localized in the cytoplasm of the tumor cells but absent from the surrounding stromal tissue. CYP1B1 was also detected in premalignant prostatic intraepithelial neoplasia (n = 2, 100%), as well as noncancerous tissues, including benign prostatic hyperplasia (n = 27, 82%), metaplastic prostatic urothelium (n = 8, 100%), and hyperplastic prostatic urothelium (n = 14, 100%). Higher CYP1B1 protein expression in bladder vs. prostate carcinoma was confirmed by their corresponding average normalized absorbances (+/- standard deviation), measured as 1.40 +/- 0.44 and 0.55 +/- 0.09, respectively. Overall CYP1B1 staining intensity in prostate carcinoma was similar to that in prostatic intraepithelial neoplasia, benign prostatic hyperplasia, and hyper-/metaplastic urothelial tissue. No CYP1B1 was detected in normal prostate tissue. CONCLUSIONS: CYP1B1 is overexpressed in prostate carcinoma at a high frequency and is also detectable in the associated premalignant and hyperplastic tissue, implicating a possible link with malignant progression and CYP1B1 as a suitable target for therapy. Spectral imaging microscopy has highlighted differences in CYP1B1 protein expression between different cancers. PMID- 14751522 TI - Repopulation of moderately well-differentiated and keratinizing GL human squamous cell carcinomas growing in nude mice. AB - PURPOSE: It has been suggested that, reminiscent of the regulated proliferative response of normal squamous epithelium, squamous cell carcinomas that have preserved characteristics of differentiation have a greater repopulation capacity during fractionated irradiation than undifferentiated tumors. The aim of the present study was to investigate repopulation in moderately well-differentiated and keratinizing GL human squamous cell carcinomas in nude mice. METHODS AND MATERIALS: GL human squamous cell carcinomas were transplanted s.c. into the right hind leg of NMRI nu/nu mice. Irradiation was performed with 5.4 Gy fractions under clamp hypoxia or with 2 Gy fractions under ambient conditions. Six, 12, or 18 fractions were given daily, every second day, or every third day. Graded top-up doses were applied under clamp hypoxia to determine the tumor control dose 50% (TCD(50)). A total of 20 TCD(50) assays were performed and analyzed using maximum-likelihood techniques. RESULTS: With an increasing number of daily 5.4 Gy fractions under clamp hypoxia, the top-up TCD(50) values decreased significantly from 50.9 Gy (95% CI: 47, 54) after single doses to 0 Gy after 18 fractions. For the same number of fractions, the top-up TCD(50) increased with increasing overall treatment time. The results are consistent with a constant repopulation rate with a clonogenic doubling time (T(clon)) of 12.7 days (8.6, 16.8). Under ambient blood flow, the top-up TCD(50)s for daily 2 Gy fractions decreased significantly, but were less pronounced than for 5.4 Gy fractions under clamp hypoxia. For a given number of fractions under ambient conditions, the top-up TCD(50)s did not increase significantly with overall treatment time, except for irradiation with 12 fractions in 36 days compared to 12 and 24 days. The T(clon) value from these data was 24.0 days (11.6, 36.4). CONCLUSION: Our data demonstrate a slow but significant rate of repopulation of clonogenic tumor cells during fractionated irradiation of GL human squamous cell carcinomas under clamp hypoxia without indication of a change of the repopulation rate during treatment. Less pronounced repopulation was observed for irradiation under ambient conditions, which might be explained by preferential survival of hypoxic and therefore nonproliferating cells. Taken together with our previous studies on poorly differentiated FaDu tumors (Petersen et al., IJROBP 2001;51:483 493), the results support important heterogeneity of kinetics and mechanisms of repopulation, in particular of the influence of the oxygenation status of surviving clonogenic cells on the repopulation rate during fractionated irradiation, in different experimental squamous cell carcinoma. PMID- 14751523 TI - Treatment of intracranial rat glioma model with implant of radiosensitizer and biomodulator drug combined with external beam radiotherapy. AB - PURPOSE: To evaluate an intracranial polymer implant containing bromodeoxyuridine (BrdUrd) and N-(phosphonacetyl)-L-aspartic acid (PALA) in combination with external beam radiotherapy (EBRT) in the treatment of a rat glioma. METHODS AND MATERIALS: Combinations of the biomodulators 5-fluorouracil, methotrexate, or PALA with BrdUrd were evaluated as radiosensitizers in vitro by clonogenic assay. In in vivo experiments, BrdUrd and PALA were incorporated into a polyanhydride based polymer, bis(p-carboxyphenoxy)propane sebacic acid, and implanted in the C6 rat glioma growing intracranially. The effectiveness of treatment was evaluated on the basis of survival. EBRT was given as 10-MV X-rays. RESULTS: In tissue culture experiments, C6 cells were refractory to radiosensitization by BrdUrd even when the thymidine analog was combined with a biomodulator intended to reduce de novo thymidine synthesis. The most effective compound in vitro was PALA. When PALA and BrdUrd in a polymer formulation were implanted intracranially and combined with 10-Gy EBRT, the treatment was highly effective, with 83% of treated rats surviving 180 days. CONCLUSION: Although the in vitro results were not encouraging, the combination of intratumoral BrdUrd and PAL with 10-Gy EBRT was highly effective in treating a rat glioma. These results indicate the clinical potential of combined and mixed modality treatments involving intratumoral sustained-release drug delivery. PMID- 14751524 TI - 18F-FDG positron emission tomography staging and restaging in rectal cancer treated with preoperative chemoradiation. AB - PURPOSE: To assess the information supplied by FDG-PET in patients with locally advanced rectal cancer both in the initial staging and in the evaluation of tumor changes induced by preoperative chemoradiation (restaging). METHODS AND MATERIALS: Twenty-five consecutive patients with rectal cancer were included, with tumor stages (c)T(2-4)N(x)M(0), during the period 1997-1999. We prospectively performed two FDG-PET scans in all patients to assess disease stage (1) at initial diagnosis and (2) presurgically, 4 to 5 weeks after protracted chemoradiation. Protracted chemoradiation was carried out during 5-6 weeks with 45-50 Gy, plus concurrent oral tegafur 1200 mg/day or 5-fluorouracil 500-1000 mg/m(2) administered as a 24-h continuous i.v. infusion on Days 1-4 and 21-25 of the radiotherapy treatment. Tumors were staged with CT in 95% of patients, whereas endorectal ultrasound was used in 90% of patients. Maximum standardized uptake value (SUVmax) was used as the quantitative parameter to estimate the tumor:tissue metabolic ratio. RESULTS: Preoperative chemoradiation significantly decreased the SUVMAX: 5.9 (mean SUVmax at initial staging) vs. 2.4 (mean SUVmax after chemoradiation) with p < 0.001. Unknown liver metastases were detected by FDG-PET in 2 patients, in 1 of them with the initial staging FDG-PET scan, and with the restaging FDG-PET scan in the other. After an average follow-up of 39 months, the value of SUVmax > or =6 allowed us to discriminate for survival at 3 years: 92% vs. 60% (p = 0.04). T downstaging (total 62%) was significantly correlated with SUVmax changes: 1.9 vs. 3.3 (p = 0.03). The degree of rectal cancer response to chemoradiation, established as mic vs. mac categories, was not associated with SUVmax differences (mean values of 2.0 vs. 2.7). CONCLUSION: Preliminary results observed suggest the potential utility of FDG-PET as a complementary diagnostic procedure in the initial clinical evaluation (8% of unsuspected liver metastases) as well as in the assessment of chemoradiation response (any T downstaged event) of locally advanced rectal cancer. Initial SUVmax might be of prognostic value related to long-term patient outcome. PMID- 14751525 TI - In vivo tumor imaging using a near-infrared-labeled endostatin molecule. AB - PURPOSE: Endostatin is a 20-kD C-terminal fragment of collagen XVIII and is a potent inhibitor of angiogenesis. Imaging technologies that use near-infrared (NIR) fluorescent probes are well suited to the laboratory setting. The goal of this study was to determine whether endostatin labeled with a NIR probe (Cy5.5) could be detected in an animal after intraperitoneal injection and whether it would selectively localize in a tumor. METHODS: Endostatin was conjugated to Cy5.5 monofunctional dye and purified from free dye by gel filtration. LLC, a murine tumor, was implanted in C57BL/6 mice. The tumors were allowed to grow to 350 mm(2), at which point the mice were injected with 100 microg/100 microL endostatin-Cy5.5 and imaged at various points under sedation. Imaging was performed using a lightproof box affixed to a fluorescent microscope mounted with a filter in the NIR bandwidth (absorbance maximum 675 nm and emission maximum 694 nm). Images were captured by a CCD and desktop computer and stored as 16-bit Tiff files. The mice were also serially imaged for uptake into the tumor and washout from the tumor. RESULTS: After intraperitoneal injection, endostatin-Cy5.5 was quickly absorbed, producing a NIR fluorescent image of the tumors at 24 h that persisted through 7 days. However, the signal peaked at 42 h after injection. Control animals included mice containing green fluorescent protein (GFP) under the control of an actin promotor, which expresses GFP in every cell; tumor-free mice injected with endostatin-Cy5.5; mice with tumors that were not injected with endostatin-Cy5.5; and mice with tumors injected with dye alone. In the four sets of control animals, no NIR photon emissions were detected at 24 hours or 5 days. Only the GFP mouse was detected using the GFP filter. Unlike previous analogous studies with 4-N-(S-glutathionylacetyl)amino) phenylarsenoxide (GSAO)-Cy5.5 in which the tumor image faded with time, the endostatin-Cy5.5 NIR signal was emitted from the tumor up to 7 days after injection, the last time point examined. CONCLUSION: The results of this study demonstrated that endostatin covalently bound to Cy5.5 will migrate from a distant intraperitoneal injection site to a tumor. These data indicate that endostatin-Cy5.5 is appropriate for selectively imaging tumors in experimental animals. Furthermore, data suggest that the anti-angiogenic effect of endostatin occurs through a local mechanism of action, within the tumor or tumor vasculature, rather than through a systemic mechanism. PMID- 14751526 TI - Apoptosis-modulating agents in combination with radiotherapy-current status and outlook. AB - PURPOSE: To increase the therapeutic efficacy of ionizing radiation or to reduce radiation-mediated side effects, diverse research centers for translational radiation oncology have headed for a specific modulation of defined cellular death pathways. In this regard, several signaling systems have proved to be of high potential value. RESULTS: It has previously been shown that apoptotic pathways induced by ionizing radiation are distinct from death pathways triggered by death ligands such as tumor necrosis factor-related apoptosis-inducing ligand (TRAIL). The combination of both radiation and TRAIL was highly efficient in vitro and in preclinical mouse models. However, several aspects of normal tissue toxicity have not been solved, and no Phase I data are available yet. A second approach tested in a Phase I trial is based on the observation that synthetic phospholipid derivatives (alkyllysophospholipids and alkylphosphocholines) strongly enhance apoptotic effects by modulating the balance among the mitogenic, anti-apoptotic MAPK, phosphatidylinositol 3'-kinase (PI3K)/Akt, and the pro apoptotic SAPK/JNK signaling pathways. Furthermore, others have provided evidence that inhibition of anti-apoptotic signals generated by mitogenic stimuli may increase radiation responses. In this context, controversial data are available regarding the influence of a pharmacologic abrogation of MEK1, Erk1/2 signaling on apoptotic sensitivity but no Phase I trials of MEK inhibitors either alone or in combination with radiation have yet been published. However, inhibition of the PI3K/Akt survival pathway using compounds such as the protein kinase C (PKC) inhibitor PKC412 has been shown to induce apoptosis or to increase the apoptotic sensitivity of tumor cells. Therefore, these drugs may be used alone or in combination with radiation to increase tumor control; however, Phase I data are lacking. Several other drugs, including cyclooxygenase-2 inhibitors, betulinic acid, and proteasome inhibitors, have been shown to interact with apoptotic signal transduction. Again, most of the drugs have not been tested in combination with radiation in vivo or-in the case of cyclooxygenase-2 inhibitors-exert pleiotropic effects. CONCLUSION: Although the examples do not reflect all available strategies, it is clear that several promising approaches targeting defined cell death pathways have been developed and entered into clinical trials. The use of synthetic phospholipid derivatives in a Phase I trial is an important example, proving that basic research in radiation biology finally guides the development of new treatment strategies. This, and other approaches, will hopefully increase tumor control rates and reduce side effects in the future. PMID- 14751527 TI - Predicting response to treatment in human cancers of the uterine cervix: sequential biopsies during external beam radiotherapy. AB - PURPOSE: To quantify cervix tumor response via weekly biopsies during therapy. METHODS AND MATERIALS: Pretreatment and subsequent weekly biopsies (when feasible) were obtained during radical external beam radiotherapy, reduced to single-cell suspensions, and assessed for DNA content by flow cytometry. RESULTS: Data for 41 patients ranging from FIGO Stage Ib to IIIb and with follow-up >100 days are presented. No clinically significant complications due to the weekly biopsies were noted. The actuarial progression-free survival at 2 years was 68%, with initial pelvic progression observed in 3 patients. The biopsies showed (1) Increased tumor cell proliferation was evident even for the earliest on-treatment samples; (2) Sustained cell yields, coupled with high proliferation 2 or more weeks into treatment, were associated with early failure; and (3) Therapy often "selected" for cell subpopulations that were only minor components of the pretreatment biopsy. CONCLUSIONS: "Accelerated repopulation" began surprisingly rapidly in cervix tumors after the initiation of chemoradiotherapy, and patients with a sustained cell yield and S-phase fraction 2 or more weeks into therapy were at increased risk for tumor progression. Although clearly implicating tumor (re)growth kinetics as a factor in outcome, the sequential biopsy data also suggested interplay between growth potential and in situ sensitivity to treatment. PMID- 14751528 TI - The prognostic value of epidermal growth factor receptor is related to tumor differentiation and the overall treatment time of radiotherapy in squamous cell carcinomas of the head and neck. AB - Accelerated repopulation in head-and-neck carcinomas might be related to the expression of proliferative factors such as epidermal growth factor receptor (EGFr). The present study focuses on the prognostic value of EGFr for T-site control and the relation to tumor cell differentiation and overall treatment time. We studied 336 patients treated with primary radiotherapy using 66-68 Gy, 2 Gy per fraction and overall treatment times of 912, 612, or 512 weeks. Pretreatment biopsies were stained for EGFr.Thirty-five percent of the carcinomas had less than 50% of the area stained for EGFr. Small T-size and well differentiated tumors was associated with a high degree of staining (p = 0.001 and p = 0.002, respectively). EGFr was of poor prognostic influence regarding local control in patients treated with 9 weeks split-course, whereas the opposite was found for patients given accelerated treatment in 5 weeks. A similar relationship between outcome, overall treatment time, and differentiation has previously been shown. The two parameters were analyzed together by separating the tumors with low EGFr and/or poor differentiation from tumors with well/moderate differentiation and high EGFr, resulting in odds ratios for T-site failure of 12 (1.43-104), 0.91 (0.51-1.65), and 0.43 (0.17-1.08), for treatment times of 912, 612, and 512 weeks, respectively. The tumor response to variations in fractionation is heterogeneous, and the prognostic impact of EGFr and differentiation might be relative and dependent on the overall treatment time of radiotherapy. PMID- 14751529 TI - Effects of low-dose ionizing radiation on gene expression in human skin biopsies. AB - PURPOSE: Several investigations have demonstrated that significant biologic effects can occur in animals, animal cells, immortalized human cell lines, and primary human cells after exposure to doses of ionizing radiation in the low dose, < or =1-10 cGy region (LDIR). However, little information is available as to how these and other observations pertain to human responses to LDIR, though such knowledge is required for reducing the uncertainty of assessing human risks due to these exposures. We therefore undertook these translational studies to begin the development of a unique data set of human cellular responses to LDIR as measured by gene expression changes when exposure occurs to a normal tissue with its complex cellular mixture and three-dimensional architecture. METHODS AND MATERIALS: Using full-thickness human skin resected during esthetic surgery, we obtained biopsy cores and exposed the tissue to LDIR ex vivo. Gene expression changes in five core regulatory genes were assessed by real-time RT-PCR. RESULTS: Results indicate that skin is a good biologic model for assessing LDIR in humans, though meticulous attention to sample processing is necessary. LDIR does produce changes in gene expression, though time- and dose-response relationships may be complex. CONCLUSION: These proof-of-principle studies have provided a crucial initial step toward validation of LDIR risk assessment models in humans. We have demonstrated the feasibility of this approach and provide initial evidence that ionizing radiation exposures as low as 1 cGy are biologically active in human skin. PMID- 14751530 TI - Radiotherapy and the potential exploitation of bystander effects. AB - Radiation-induced bystander effects are the subject of intense investigation in radiation protection. The effects predominate at low doses and have been discussed mainly in terms of the impact on low-dose risk assessment. Possible therapeutic implications have been alluded to, but not discussed in any detail. The purpose of this review was to consider bystander biology in areas of major importance or interest in radiotherapy. These include consideration of radiation induced bystander effects during the cell cycle, under hypoxic conditions, when fractionated therapy modalities are used, or when combined radiochemotherapy is given. Also discussed are individual variations in toxicity of bystander factors and normal tissue "collateral" damage. The importance of considering the tumor in the context of the organ, and even the organism that supports it, is also discussed. Direct clinical radiotherapy studies that consider bystander effects are not in the public domain at the time of writing, but many in vitro studies are available that are relevant; some preliminary animal data have also been published. Because radiation-induced bystander effects appear to challenge many of the central assumptions that underlie radiotherapy practice, it is important to consider what unexplored treatment avenues might result from a consideration of these effects. The final part of this paper is devoted to this point. PMID- 14751531 TI - A bispecific antibody to enhance radiotherapy by tumor necrosis factor-alpha in human CEA-expressing digestive tumors. AB - Tumor necrosis factor-alpha (TNF-alpha) enhances X-ray killing of human tumor cells in vitro and enhances tumor control when combined with radiotherapy (RT) in animal tumor models. In multiple Phase I studies, intravenous injection of TNF alpha appeared to have severe systemic side effects. To overcome these limitations, we used a bispecific antibody (BAb) directed against carcinoembryonic antigen and human TNF-alpha to target this cytokine in human digestive carcinoma treated with simultaneous RT. We used human digestive carcinoma cell lines (colon cancer, LS174T, and pancreatic cancer, BxPC-3) to determine the interaction of TNF-alpha and RT on clonogenic cytotoxicity. Isobolograms were established to confirm additive or supra-additive effects between both treatments. LS174T and BxPC-3 cells were grafted subcutaneously at Day 0 into female nude mice (7-8 weeks old). When the tumors reached a volume of about 80 mm(3), the mice were randomly assigned to treatment: Group 1, normal saline i.v. injection (control group); Group 2, TNF-alpha at 1 microg/i.v. injection; Group 3, BAb at 25 microg/i.v. injection; Group 4, BAb plus TNF-alpha (ratio 25 microg to 1 microg) i.v. injection; Group 5, local RT plus normal saline (0.5 Gy. min(-1)) at a total dose of 30 Gy delivered in five fractions; Group 6, local RT plus TNF-alpha injections 3 h before RT; Group 7, local RT plus BAb plus TNF-alpha co-injected 24 h before RT. Tumor growth delay was used as the end point for all groups. In the LS174T experiments, TNF-alpha added 12 h before RT showed a statistically significant decrease in the survival fraction at 2 Gy compared with RT alone (0.23 vs. 0.42 Gy, p = 0.0017). These results were largely confirmed with the BxPC-3 cell lines (0.29 vs. 0.72, p <0.00001). Isobolograms confirmed the additivity between TNF-alpha and RT in both cell lines. At 50% survival, the data points were within the envelope of additivity. In the LS174T and BxPC-3 xenografts, RT as a single agent (Group 5) slowed tumor progression compared with Group 1 (p <0.027 and p = 0.00001, respectively). TNF-alpha alone, BAb alone, or BAb plus TNF-alpha (Groups 2, 3, and 4) had no effect. In the LS174T model, TNF-alpha plus RT enhanced the delay to reach 2000 mm(3) compared with RT alone but without statistical significance. This delay was significantly longer when BAb was added (p = 0.0033, for Group 6 vs. Group 7). In the BxPC-3 experiments, the median delay to reach 2000 mm(3) was similar between the RT and TNF-alpha plus RT groups (93 days). The use of our BAb in combination with TNF alpha and RT dramatically enhanced this median delay (177 days, p = 0.0013). No body weight loss was observed in any group. Our data could be used as a solid preclinical rationale on which to base a clinical study of locally advanced pancreatic or rectal cancers in the near future. PMID- 14751532 TI - Relationships between radiosensitivity and microvascular density in esophageal carcinoma: significance of hypoxic fraction. AB - PURPOSE: The prognosis and the radiosensitivity of macroscopically infiltrative type of esophageal carcinoma are worse than those of the localized type of esophageal carcinoma treated with irradiation. The aim of this study was to investigate the cause of differences in radiosensitivity and prognosis of esophageal carcinoma according to macroscopic type from the viewpoint of tumor angiogenesis. METHODS AND MATERIALS: A total of 40 surgically resected esophageal carcinoma tissues with good material remaining were selected at random from macroscopically localized type (n = 20) and infiltrative type (n = 20) of esophageal carcinoma. The highest intratumoral microvascular density (h-MVD), average intratumoral microvascular density (a-MVD), Ki67 labeling index, and expression of vascular endothelial growth factor (VEGF) in each section were estimated. RESULTS: h-MVD was significantly (p = 0.0006) greater in the infiltrative type than in the localized type, whereas a-MVD (p = 0.0014) and Ki67 labeling index (p = 0.022) were significantly lower in the infiltrative type than in the localized type. The expression level of VEGF was significantly (p < 0.0001) higher in the infiltrative type. CONCLUSIONS: The generally underdeveloped vascular densities with low proliferation activities (suggesting increase of hypoxic fraction) seemed to be one of the reasons for unfavorable radiosensitivities of infiltrative type of esophageal carcinoma. The infiltrative type of esophageal carcinoma shows a high level of VEGF expression and high activity of tumor angiogenesis. The locally enhanced neovascularization, which occurs frequently in hematogenous metastasis seemed to be one of the reasons for the unfavorable prognosis of the infiltrative type of esophageal carcinoma. PMID- 14751533 TI - Where next with preoperative radiation therapy for rectal cancer? AB - PURPOSE: The basic question for radiation oncologists is what we hope to achieve from treatments that are adjuvant to surgery: better local (pelvic) control and, hopefully, because of that, fewer metastases. Chemotherapy could add to the local effect of irradiation and may also decrease distant metastases directly. Selection criteria for individual treatment could enhance the therapeutic index. LOCAL CONTROL: Total mesorectal excision reduces the incidence of local recurrence, but preoperative (chemo) radiation is still indicated for more advanced tumors (T3-T4) and for lymph node involvement. Pelvic recurrences arise from tumor clonogens residual beyond the surgical margins. Thus, the practice of shrinking fields to boost the dose to the primary tumor makes no sense, except for tumors that invade residual structures, such as the sacrum. Subclinical disease beyond the future surgical margins grows more quickly than the primary tumor, and hence treatment should be as intense as tolerable. A short treatment course (e.g. 5 x 5 Gy) is desirable, but this regimen, which is currently the gold standard, should be compared (as in the recently closed randomized Polish trial) with higher-dose, longer-duration chemoradiotherapy regimens. The recently closed EORTC trial 22921 examines the benefit of pre- and postoperative chemotherapy combined with a long schedule of radiation. Likewise, continuous infusion of a cycle-active agent rather than bolus administration is a logical addition to radiation therapy in the treatment of fast-growing subclinical tumor extensions. SYSTEMIC DISEASE: The reduction in distant metastasis rates attributable to adjuvant chemotherapy varies greatly among reports. If the reduction is of the order of 10-25%, the efficacy of chemotherapy equates to as little as about 5 to 12.5 Gy and not more than 20 Gy of total body irradiation. INTERVAL BETWEEN RADIATION THERAPY AND POSTRADIATION SURGERY: Early excision after preoperative irradiation would be desirable if the primary tumor were still disseminating viable metastatic clonogens. Most tumors do not metastasize until they contain enough viable clonogens to render them clinically detectable. A dose of 10 Gy in 2 Gy fractions reduces at least 30-fold the absolute number of viable clonogens in the primary tumor, to levels that do not yield metastases from the untreated tumor. After a dose of 44-50 Gy in 2 Gy fractions, there is little chance that the surviving tumor clonogens could regrow to a metastasis-yielding volume in any reasonable radiation-surgery interval. Thus there is no tumor related necessity for early postradiation surgery. The importance of the interval between radiation and surgery is currently being addressed in a Swedish randomized trial. PROGNOSTIC AND PREDICTIVE CHARACTERIZATION: Tumor volume should be included in the staging system. There are many tumor- and host-related characteristics that can be used to fingerprint the tumor to help select appropriate individual treatment. PMID- 14751534 TI - Recent advances in light ion radiation therapy. AB - BACKGROUND: The fast development of energy- and intensity-modulated radiation therapy during the last two decades using photon and electron beams has when implemented resulted in a considerable improvement of radiation therapy, particularly if combined with radiobiologically based treatment optimization techniques. This has made intensity-modulated electron and photon beams as powerful as today's uniform dose proton therapy. To be able to cure also the most advanced hypoxic and radiation-resistant tumors of complex local spread, intensity-modulated light ion beams are really the ultimate tool and in clinical practice 2 to 3 times less expensive per patient treated than proton therapy. This development and the recent development of advanced tumor diagnostics based on PET-CT imaging of the tumor cell density open the field for new powerful radiobiologically based treatment optimization methods. The ultimate step is to use the unique radiobiologic and dose distributional advantages of light ion beams for truly optimized bioeffect planning where the integral three-dimensional dose delivery and tumor cell survival can be monitored by PET-CT imaging and corrected by biologically based adaptive therapy optimization methods. PURPOSE: The main purpose of the present paper is to discuss the principal areas of development of therapy optimization, by considering the therapy chain from tumor diagnostics and the use of three-dimensional predictive assay to biologically based treatment optimization with special focus on the rapid clinical development of advanced light ion therapy. METHODS: Besides the "classical" approaches using low ionization density hydrogen ions (protons, but also possibly deuterons and tritium nuclei) and high ionization density carbon ions, two new approaches will be discussed. In the first one, lithium or beryllium or boron ions, which induce the least detrimental biologic effect to normal tissues for a given biologic effect in a small volume of the tumor, will be key particles. In the second approach, referred patients will be given a high-dose, high-precision "boost" treatment with carbon or oxygen ions during 1 week preceding the final treatment with conventional radiation in the referring hospital. The rationale behind these approaches is to minimize the high ionization density dose to the normal-tissue stroma outside but sometimes also inside the tumor bed and to ensure a more uniform and optimal biologic effectiveness in the tumor, also on the microscopic scale. The present discussion indicates that BIologically Optimized predictive Assay based light ion Radiation Therapy (Bio-Art) is really the ultimate way to perform high-precision radiation therapy using checkpoints of the integral dose delivery and the tumor response and, based on this information, perform compensating corrections of the dose delivery. By using biologically optimized scanned high-energy photon or ion beams, it is possible to measure in vivo the three-dimensional dose delivery using the same PET-CT camera that was used for diagnosing the tumor spread. This method thus opens up the door for truly three dimensional biologically optimized adaptive radiation therapy, where the measured dose delivery to the true target tissues can be used to fine-adjust the incoming beams, so that possible errors in the integral therapy process are eliminated toward the end of the treatment. Interestingly enough, practically all major error sources--such as organ motion, treatment planning errors, patient setup errors, and dose delivery problems due to gantry, multileaf, or scanning beam errors--can be corrected for in this way. RESULTS AND CONCLUSIONS: Radiobiologically optimized dose delivery using intensity and radiation quality modulation based on high-resolution PET-CT or Magnetic Resonance Spectroscopic Imaging (MRSI)-based tumor and normal-tissue imaging is probably the ultimate development of radiation therapy, taking the unique physical and biologic advantages of light ions fully into account in truly patient-individualized curative treatment schedules. Using recently available biologically based treatment ilable biologically based treatment optimization algorithms, it is possible to improve the treatment outcome for advanced tumors by as much as 10 40%. The adaptive radiotherapy process based both on three-dimensional tumor cell survival and dose delivery monitoring has the potential of percent accuracy in tumor response and dose delivery monitoring, using two-dimensional, narrow high energy photon beam scanning and three-dimensional (11)C Bragg peak scanning for radiation quality and intensity-modulated dose delivery. There is no doubt that the future of radiation therapy is very promising, and gradually more and more patients may not even need advanced surgery. Instead, they could be cured by biologically optimized electron, photon, or light ion therapy, where the densely ionizing Bragg peak is placed solely in the gross tumor, and a lower ionization density is used in microscopically invasive tumor volumes. PMID- 14751535 TI - Three-dimensional conformal vs. intensity-modulated radiotherapy in head-and-neck cancer patients: comparative analysis of dosimetric and technical parameters. AB - BACKGROUND AND PURPOSE: The use of intensity-modulated radiotherapy (IMRT) is now widely advocated for the treatment of head-and-neck cancers, to increase the therapeutic ratio of radiotherapy used as sole modality of treatment or in combination with chemotherapy. This report aims to summarize the technical and dosimetric factors to be taken into consideration to assess the respective advantages of the various high conformality treatments in radiotherapy, especially in the framework of quality assurance procedures. MATERIALS AND METHODS: Twenty-six head-and-neck cancer patients were irradiated following a feasibility internal protocol with IMRT. Treatments were performed with either the static step-and-shoot (20) or the dynamic sliding window (6) techniques on a 6 MV Varian Clinac equipped with a multileaf collimator with 80 leaves. Dose plans were computed using commercial treatment planning systems: MDS-Nordion Helax-TMS for static cases and Varian Eclipse for dynamic cases. Dose plans were evaluated in terms of physical quantities based on dose-volume histograms and isodose distributions. Each IMRT plan was also compared to a reference 3D conformal therapy plan (3DCRT). RESULTS: Elective target volumes ranged from 530 to 1151 cm(3) with a mean of 780 +/- 141 cm(3). Boost volumes ranged from 248 to 832 cm(3) with a mean of 537 +/- 165 cm(3). Thirty-two dose plans were generated with static technique and 10 with dynamic. In the static mode, 6.8 +/- 3.4 fields were applied on average with 12.5 +/- 1.3 segments per field. In the static mode, 264 +/- 56 MU per Gy were erogated, whereas in the dynamic mode, 387 +/- 126 MU per Gy were erogated, to be compared to 147 +/- 20 computed for reference 3DCRT plans. For all target volumes in general, conformity was improved compared to 3DCRT (e.g. V(95) increased from 85% to 93% with p < 0.001, or equivalent uniform dose normalized to prescribed dose increased from 0.86 to 0.96 with p = 0.002). Irradiation of parotid glands or spinal cord improved, as well: For parotids, D(2/3V) reduced from 59 Gy to 41 Gy (p < 0.001). For spinal cord, D(max) reduced from about 40 Gy to about 30 Gy (p < 0.001). PMID- 14751536 TI - Rapid involution and mobility of carcinoma of the cervix. AB - PURPOSE: To quantitatively describe the involution and mobility of carcinoma of the cervix while under treatment with chemoradiotherapy (both with external beam radiation [EBRT] and high-dose-rate [HDR] intracavitary therapy). These data have implications for conformal or intensity modulated radiation therapy boost to the cervix. METHODS AND MATERIALS: Seventeen patients underwent HDR brachytherapy boost to the cervix and were evaluated by repeat clinical examinations. In most cases, 5 weekly HDR brachytherapy insertions were performed after approximately 2 to 3 weeks of the initiation of EBRT. Sequential clinical tumor sizes were recorded in the chart for each patient under treatment. Linear regression analyses were performed to analyze tumor size as a function of total dose of external beam plus brachytherapy and number of elapsed days during the treatment course. In addition, the mobility of the cervix was documented by placement of a uterine sleeve for HDR brachytherapy before the initiation of therapy, and changes in sleeve position were identified on portal films relative to the midline of the pubic symphysis, in three dimensions. The anatomic position of the cervix was also identified at the time of simulation for HDR brachytherapy. RESULTS: Seventeen patients were identified and selected to receive HDR brachytherapy at our institution. Sixteen of the 17 patients received concurrent chemotherapy. The median dose at which tumor was no longer clinically evident was 61.5 Gy (95% confidence interval [CI]: 50.7-72.3 Gy) by linear regression analysis. This indicates that the median dose to achieve a 50% reduction in tumor size is approximately 30.8 Gy. Similarly, the median number of elapsed days for a complete response was 42 days (95% CI: 34-50 elapsed days). This indicates that it takes 21 days to achieve a 50% clinical complete response for patients undergoing concurrent cisplatin-based chemoradiotherapy and HDR brachytherapy. In addition, the mobility of the cervix during EBRT was noted by serial measurements of the location of a metallic ring in the uterine sleeve, as seen on port films. The median and maximum ranges for change in the position of the cervix in the lateral (x), superior/inferior (y), and anterior/posterior (z) planes were 10, 8, and 16 mm and 24, 36, and 23 mm, respectively. Also, 85 brachytherapy procedures were performed, and the positions of the cervix on 170 orthogonal films were evaluated. The median and maximum ranges for the position of the cervix at the time of HDR brachytherapy in the lateral (x), superior/inferior (y), and anterior/posterior (z) planes were 5, 12, and 10 mm and 11, 25, and 32 mm, respectively. CONCLUSIONS: Carcinoma of the cervix involutes rapidly with EBRT, concurrent cisplatin-based chemotherapy, and HDR brachytherapy. The time for 50% tumor regression was calculated to be 21 days and occurs after 30.8 Gy. In addition, uterine sleeve placement allowed us to document the median and maximum ranges of cervical mobility during the treatment course of EBRT to be 8-16 mm and 23-36 mm, and at the time of HDR brachytherapy to be 5-12 mm and 11-32 mm, respectively. These data indicate that the cervix gross tumor volume changes rapidly in a systematic fashion during chemoradiotherapy and, together with the mobility of the cervix, urge caution in nonbrachytherapy boost planning. PMID- 14751537 TI - Results of carbon ion radiotherapy in 152 patients. AB - PURPOSE: This study summarizes the experience with raster scanned carbon ion radiation therapy (RT) at the Gesellschaft fur Schwerionenforschung (GSI), Darmstadt, Germany since 1997. METHODS AND MATERIALS: Between December 1997 and December 2002, 152 patients were treated at GSI with carbon ion RT. Eighty-seven patients with chordomas and low-grade chondrosarcomas of the skull base received carbon ion RT alone (median dose 60 GyE); 21 patients with unfavorable adenoid cystic carcinomas and 17 patients with spinal (n = 9) and sacrococcygeal (n = 8) chordomas and chondrosarcomas were treated with combined photon and carbon ion RT. Twelve patients received reirradiation with carbon ions with or without photon RT for recurrent tumors. Furthermore, 15 patients with skull base tumors other than chordoma and low-grade chondrosarcoma were treated with carbon ions. RESULTS: Actuarial 3-year local control was 81% for chordomas, 100% for chondrosarcomas, and 62% for adenoid cystic carcinomas. Local control was obtained in 15/17 patients with spinal (8/9) and sacral (7/8) chordomas or chondrosarcomas and in 11/15 patients with skull base tumors other than chordomas and low-grade chondrosarcomas, respectively. Six of 12 patients who received reirradiation are still alive without signs of tumor progression. Common Toxicity Criteria Grade 4 or Grade 5 toxicity was not observed. CONCLUSION: Carbon ion therapy is safe with respect to toxicity and offers high local control rates for skull base tumors such as chordomas, low-grade chondrosarcomas, and unfavorable adenoid cystic carcinomas. PMID- 14751538 TI - Modeling killing and repopulation kinetics of subclinical cancer: direct calculations from clinical data. AB - PURPOSE: Models for cell killing and repopulation can provide insight into the efficacy of therapies. Using clinical data on breast cancer recurrence after lumpectomy with or without radiotherapy (L+/-RT) and brain metastases after chemotherapy with or without prophylactic cranial irradiation (C+/-PCI) for small cell lung cancer, estimates of cell killing and subclinical repopulation were tested against the results from simple radiobiologic models. METHODS AND MATERIALS: The rates of local breast cancer recurrence after L+/-RT and of brain metastases after C+/-PCI were extracted from published randomized trials. In Method 1, assuming simple exponential growth, the cell number distributions after L+/-RT and C+/-PCI were calculated from the clinical data, and the impact of RT on these distributions was determined. In Method 2, "classic" radiobiology dictates that a typical course of breast RT and PCI results in approximately =7 and approximately =4.5 log of cell kill, respectively. Using an assumption of uniform log-kill, the clinical doubling times (CDTs) can be calculated directly from the clinical data. RESULTS: Using Method 1, for breast cancer and assuming a CDT of 110 days and a clinically detectable cell number of 10(9), the calculated cell number distribution would be approximately uniformly distributed from 1 to 10(8) cells, with RT reducing the frequency at all points by approximately =75%. From the brain metastasis data, assuming a CDT of 55 days, a cell number distribution of 10(3) to 10(8) cells would be calculated. PCI reduces the frequency of metastases by roughly 40%. For both the breast and the brain data, the effects of RT on the cell number distribution are not consistent with uniform radiosensitivity. Using Method 2, assuming a cell number of 10 after L+/-RT, the calculated CDTs range from 14 to 124 days. For the brain metastasis case, assuming a starting cell number of 3.16 x 10(3), the CDTs would primarily be in the 10-30-day range. CONCLUSION: The distribution of clinical responses to adjuvant RT suggests a broad range of radiosensitivity, rather than uniform log cell kill. The subpopulation of tumors with minimal cell kill appears to be significant. This heterogeneity may be due to radioresistant subpopulations, failure to irradiate tumor cells, and/or new tumor formation. Similarly, the computed CDTs consistent with the clinical data are shorter than those reported in the literature. Simple radiobiologic models that fail to incorporate heterogeneity of radiosensitivity and/or tumor cell repopulation do not adequately describe clinical outcomes. PMID- 14751539 TI - Cell signalling through thromboxane A2 receptors. AB - Thromboxane A2 receptors (TPs) are widely distributed among different organ systems and have been localized on both cell membranes and intracellular structures. Following the initial cloning of this receptor class from human placenta, the deduced amino acid sequence predicted seven-transmembrane spanning regions, four extracellular domains and four intracellular domains, making TP a member of the seven-transmembrane G-protein-coupled receptor (GPCR) super family. A single gene on chromosome 19p13.3 leads to the expression of two separate TP isoforms: TPalpha which is broadly expressed in numerous tissues, and a splice variant termed TPbeta which may have a more limited tissue distribution. Mutagenesis, photoaffinity labelling, and immunological studies have indicated that the ligand binding domains for this receptor may reside in both the transmembrane (TM) and extracellular regions of the receptor protein. In addition, separate studies have provided evidence that this receptor can couple to at least four separate G protein families. As a consequence, TP signalling has been shown to result in a broad range of cellular responses including phosphoinositide metabolism, calcium redistribution, cytoskeletal arrangement, integrin activation, kinase activation, and the subsequent nuclear signalling events involved in DNA synthesis, cell proliferation, cell survival and cell death. While activation of these different signalling cascades can all derive from TP stimulation, the relative signalling preference for a given cascade appears to be both tissue and cell specific. Finally, separate studies have indicated that TP signalling capacity can be both down-regulated by protein kinase activation and up-regulated by GPCR cross-signalling. Thus, the multitude of signalling events which derive from TP activation can themselves be modulated by endogenous cellular messengers. PMID- 14751540 TI - Bradykinin-stimulated p42/p44 MAPK activation associated with cell proliferation in corneal keratocytes. AB - Bradykinin (BK) is released into the tear-film in ocular allergic patients. BK has been shown to exert mitogenic effects on several cell types. However, the mechanisms underlying its action on corneal keratocytes (CKs) were largely unknown. This study was to investigate the mitogenic effect of BK on rabbit CKs linked to activation of p42/p44 mitogen-activated protein kinase (MAPK), assessed by [3H]thymidine incorporation and Western blotting analysis, respectively. BK stimulated [3H]thymidine incorporation and p42/p44 MAPK phosphorylation in a time and concentration-dependent manner. Pretreatment with pertussis toxin attenuated the BK-induced responses. BK-stimulated responses were attenuated by inhibitors of selective B2 receptor (Hoe 140), phosphatidylinositol (PI)-PLC (U73122), an intracellular Ca2+chelator (BAPTA/AM), PKC (GF109203X), tyrosine kinase (genistein), and MEK1/2 (PD98059). BK also stimulated translocation of p42/p44 MAPK into nucleus and led to expression of c-fos and c-jun in CKs. These results demonstrate that in CKs, BK-stimulated phosphorylation of p42/p44 MAPK is mediated through the activation of BK B2 receptors and leads to cell proliferation. PMID- 14751541 TI - CaM kinase II-dependent phosphorylation of myogenin contributes to activity dependent suppression of nAChR gene expression in developing rat myotubes. AB - During development of the neuromuscular junction (NMJ), extrajunctional expression of genes, whose products are destined for the synapse, is suppressed by muscle activity. One of the best-studied examples of activity-dependent gene regulation in muscle are those encoding nicotinic acetylcholine receptor (nAChR) subunits. We recently showed that nAChR gene expression is inhibited by calcium/calmodulin-dependent protein kinase II (CaMKII) and CaMKII inhibitors block activity-dependent suppression of these genes. Here we report results investigating the mechanism by which CaMKII suppresses nAChR gene expression. We show that the muscle helix-loop-helix transcription factor, myogenin, is necessary for activity-dependent control of nAChR gene expression in cultured rat myotubes and is a substrate for CaMKII both in vitro and in vivo. CaMKII phosphorylation of myogenin is induced by muscle activity and this phosphorylation influences DNA binding and transactivation. Thus we have identified a signaling mechanism by which muscle activity controls nAChR gene expression in developing muscle. PMID- 14751542 TI - PKA-dependent activation of PKC, p38 MAPK and IKK in macrophage: implication in the induction of inducible nitric oxide synthase and interleukin-6 by dibutyryl cAMP. AB - In this study, we examined the signal transduction of dibutyryl cyclic adenosine monophosphate (dBcAMP) to stimulate the release of nitric oxide (NO) and interleukin-6 (IL-6) from J774 macrophages. These actions of dBcAMP were diminished by the presence of the inhibitors of protein kinase A (PKA), protein kinase C (PKC), p38 MAPK and nuclear factor-kappa B (NF-kappaB). In contrast, Go 6976 and PD98059 had no significant effects. Consistently, dBcAMP caused membrane translocation of PKCbetaII, delta, mu, lambda and zeta isoforms, and increased atypical protein kinase C (aPKC) and p38 MAPK activities. The nuclear translocation and DNA-binding study revealed that dBcAMP stimulated NF-kappaB, activator protein-1 (AP-1), and CAAT/enhancer-binding protein (c/EBPbeta). Via PKA, PKC and p38 MAPK-dependent signals, dBcAMP also induced inhibitory subunit of NF-kappaB (IkappaB) degradation, IkappaB kinase (IKK) activation, nuclear translocation of NF-kappaB subunit p65 and its association with the CREB-binding protein (CBP). These results illustrate that PKA activation in macrophages is able to stimulate PKC and p38 MAPK, which lead to IKK-dependent NF-kappaB activation and contribute to the induction of inducible nitric oxide synthase (iNOS) and IL-6 genes. PMID- 14751543 TI - Induction of Cbeta splice variants and formation of novel forms of protein kinase A type II holoenzymes during retinoic acid-induced differentiation of human NT2 cells. AB - Cyclic AMP (cAMP) and cAMP-dependent protein kinase (PKA) are critical regulators of neuronal differentiation. The expression, levels and activities of PKA subunits were studied prior to and during differentiation of the human neuronal precursor cell line NTera 2 (NT2). Undifferentiated NT2 cells expressed mainly cytoplasmic PKA type I, consisting of the regulatory subunit RIalpha and the catalytic subunit Calpha. Low levels of PKA type II consisting of RIIalpha or RIIbeta associated with Calpha were also detected, mainly in the cytoplasm and in the Golgi-centrosomal area. During retinoic acid-induced differentiation, the RIalpha and RIIalpha expressions remained in the cytoplasm, while we observed a strong upregulation of RIIbeta, located to the whole cytoplasm including neurite extensions. This upregulation coincided with increased PKA-specific activity accompanied by a strong induction of a number of neuronal-specific Cbeta splice variants that together with RIIbeta form novel PKAII holoenzymes. Formation of novel PKAII holoenzymes may imply specific PKA features which may have consequences for the process of neuronal differentiation and nerve cell function. PMID- 14751544 TI - Phosphorylation of profilin regulates its interaction with actin and poly (L proline). AB - Activation of bovine platelets with thrombin and phorbol 12,13-dibutyrate (PDBu) resulted in phosphorylation of profilin on serine. The phosphorylation was inhibited when platelets were pretreated with the PI 3-kinase inhibitor, LY294002, indicating that profilin phosphorylation is a downstream event with respect to PI 3-kinase activation. Phosphorylation of profilin resulted in significant decrease in actin polymerization (16.5%), indicating an increased affinity of phosphoprofilin towards actin. The critical actin monomer concentration (Cc) increased to 260 nM in the presence of phosphoprofilin in comparison with 200 nM in the presence of profilin. The interaction of phosphoprofilin with phosphatidylinositol 4,5-bisphosphate [PI (4,5)-P2] and poly (L-proline) (PLP) was examined by monitoring the quenching of tryptophan fluorescence. Scatchard plot and binding isotherm data obtained revealed no difference in PI (4,5)-P2 binding between profilin and phosphoprofilin (Kd=20.4 microM), while poly (L-proline)-binding studies indicated a sixfold decrease (27.34 microM for profilin and 4.73 microM for phosphoprofilin) in Kd with phosphoprofilin. In vivo studies with platelets indicated an increased association of p85alpha, the regulatory subunit of PI 3-kinase with phosphoprofilin over profilin. Overall, the data presented conclude that profilin phosphorylated under in vivo conditions and phosphorylation depends upon activation of PI 3-kinase. Phosphoprofilin exhibited increased affinity to poly (L-proline) sequences both in vitro and in vivo. PMID- 14751545 TI - Tumor necrosis factor-alpha-induced cyclooxygenase-2 expression in human tracheal smooth muscle cells: involvement of p42/p44 and p38 mitogen-activated protein kinases and nuclear factor-kappaB. AB - This study was to determine the mechanism of tumor necrosis factor-alpha (TNF alpha)-enhanced cyclooxygenase (COX)-2 expression associated with prostaglandin E2 (PGE2) synthesis in human tracheal smooth muscle cells (HTSMCs). TNF-alpha markedly increased COX-2 expression and PGE2 synthesis in a time- and concentration-dependent manner, whereas COX-1 remained unaltered. Tyrosine kinase inhibitor (genistein), phosphatidylcholine-specific phospholipase C (PC-PLC) inhibitor (D-609) and PKC inhibitor (GF109203X) attenuated TNF-alpha-induced COX 2 expression and PGE2 synthesis in HTSMCs. TNF-alpha-induced COX-2 expression and PGE2 synthesis were also inhibited by PD98059 (an inhibitor of MEK1/2) and SB203580 and SB202190 (inhibitors of p38 MAPK), respectively, suggesting the involvement of p42/p44 and p38 MAPKs in these responses. This hypothesis was further supported by that TNF-alpha induced a transient activation of p42/p44 and p38 MAPKs in a time-and concentration-dependent manner. Furthermore, TNF-alpha induced activation of nuclear factor-kappaB (NF-kappaB) reversely correlated with the degradation of IkappaB-alpha in HTSMCs. TNF-alpha-induced COX-2 expression and PGE2 synthesis was also inhibited by NF-kappaB inhibitor pyrrolidinedithiocarbamate (PDTC). These findings suggest that the increased expression of COX-2 correlates with the release of PGE2 from TNF-alpha-challenged HTSMCs, at least in part, mediated through p42/p44 and p38 MAPKs as well as NF kappaB signaling pathways in HTSMCs. PMID- 14751546 TI - Involvement of Ca2+ channels in endothelin-1-induced MAP kinase phosphorylation, myosin light chain phosphorylation and contraction in rabbit iris sphincter smooth muscle. AB - The purpose of the present study was to investigate the role and type of Ca2+ channels involved in the stimulatory effects of endothelin-1 (ET-1) on the Ca2+ dependent functional responses, p42/p44 MAP kinase phosphorylation, 20-kDa myosin light chain (MLC) phosphorylation and contraction, in rabbit iris sphincter, a nonvascular smooth muscle. ET-1 induced inositol phosphates production, MAP kinase phosphorylation, MLC phosphorylation (MLC20-P plus MLC20-2P) and contraction in a concentration-dependent manner with EC50 values of 71, 8, 6 and 25 nM, respectively. ET-1-induced MAP kinase phosphorylation, MLC phosphorylation and contraction were not significantly affected by nifedipine (1-60 microM), an L type Ca2+ channel blocker, or by LOE 908 (1-100 microM), a blocker of Ca2+ permeable nonselective cation channels. However, SKF96365, a receptor-operated Ca2+ channel (ROCC) blocker, inhibited MAP kinase phosphorylation, MLC phosphorylation and contraction in a concentration-dependent manner with IC50 values of 28, 30 and 42 microM, respectively. 2-APB, a store-operated Ca2+ channel (SOCC) blocker, inhibited ET-1-induced MLC phosphorylation and contraction in a concentration-dependent manner with IC50 values of 12.7 and 19 microM, respectively, but was without effect on MAP kinase phosphorylation. The combined effects of submaximal concentrations of SKF96365 and 2-APB on ET-1 induced MLC phosphorylation and contraction were not additive, implying that their inhibitory actions could be mediated through a common Ca2+ entry channel. PD98059, a MAP kinase inhibitor, had no effect on ET-1-induced MLC phosphorylation and contraction, suggesting that these ET-1 effects in the rabbit iris muscle are MAP kinase-independent. In conclusion, the present study demonstrated for the first time that in rabbit iris sphincter (a) ET-1, through the ETA receptor, stimulates MAP kinase phosphorylation, MLC phosphorylation and contraction in a concentration-dependent manner, (b) that these Ca2+-dependent functional responses are not significantly affected by nifedipine or LOE908, and (c) that ET-1-induced MLC phosphorylation and contraction are inhibited by SKF96365 and 2-APB, suggesting that these effects are mainly due to store- and/or receptor Ca2+ entry. PMID- 14751547 TI - Regulation and localization of CAS substrate domain tyrosine phosphorylation. AB - Crk-associated substrate (CAS) is a tyrosine kinase substrate implicated in integrin control of cell behavior. Phosphorylation, by Src family kinases, of multiple tyrosine residues in the CAS substrate domain (SD) is a major integrin signaling event that promotes cell motility. In this study, novel phosphospecific antibodies directed against CAS SD phosphotyrosine sites ("pCAS" antibodies) were characterized and employed to investigate the cellular regulation and localization of CAS SD tyrosine phosphorylation. An analysis of CAS and focal adhesion kinase (FAK) variants expressed in CAS- and FAK-deficient cell lines, respectively, indicated that CAS SD tyrosine phosphorylation is substantially achieved by Src family kinases brought into association with CAS through two distinct mechanisms: direct binding to the CAS Src-binding domain and indirect association through a FAK bridge. Cell immunostaining with pCAS antibodies revealed that CAS SD tyrosine phosphorylation occurs exclusively at sites of integrin adhesion including both nascent focal complexes formed at the edges of extending lamellipodia as well as mature focal adhesions underlying the cell body. These findings further document a role for FAK as an important upstream regulator of CAS SD tyrosine phosphorylation and implicate CAS-mediated signaling events in promoting membrane protrusion/lamellipodium extension during cell motility. PMID- 14751548 TI - Contribution of JNK, Mek, Mos and PI-3K signaling to GVBD in Xenopus oocytes. AB - In Xenopus oocytes, induction of the G2/M transition by progesterone is a complex process that is promoted by a network of signaling molecules whose cumulative effect results in the activation of maturation promoting factor (MPF) and germinal vesicle breakdown (GVBD). We examined the role of Mos, Mek, PI-3 kinase and c-Jun N-terminal kinase (JNK) in progesterone stimulation of GVBD. Expression of an activated form of JNK neither induced nor enhanced progesterone-mediated GVBD in oocytes, suggesting a limited role in cell-cycle progression. We blocked Mek, Mos and PI-3 kinase activities by a variety of means that included expression of dominant-negative kinase suppressor of Ras (DnKSR), expression of a dominant-negative PI-3 kinase (DnPI3K), treatment of oocytes with a Mek inhibitor (U1026) or PI-3 kinase (LY294002) inhibitor, and introduction of Mos antisense morpholinos. Inhibition of any one pathway alone failed to block GVBD induced by either high or low concentrations of progesterone. In contrast, inhibiting Mos or Mek function in addition to abrogating PI-3 kinase activity effectively blocked oocyte maturation. Furthermore, by expressing suboptimal amounts of Mos in conjunction with an activated form of Mek and an activated form of the p110 catalytic subunit of PI-3 kinase, we show cooperation among these signaling molecules toward the induction of GVBD. Moreover, expression of optimal amounts of these three proteins in conjunction with inhibitors of Mos, Mek or PI-3 kinase demonstrated that activated Mek-induced GVBD is independent of Mos or PI-3 kinase activity. In addition, Mos-induced GVBD is dependent upon Mek activity, but does not require PI-3 kinase activity. Finally, Mos appears to be a major contributor to GVBD induced by activated PI-3 kinase, while Mek is a minor contributor to this process. PMID- 14751549 TI - Retinoic acid signals the direction of ocular elongation in the guinea pig eye. AB - A growing eye becomes myopic after form deprivation (FD) or compensates for the power and sign of imposed spectacle lenses. A possible mediator of the underlying growth changes is all-trans retinoic acid (RA). Eye elongation and refractive error (RE) was manipulated by raising guinea pigs with FD, or a spectacle lens worn on one eye. We found retinal-RA increased in myopic eyes with accelerated elongation and was lower in eyes with inhibited elongation. RA levels in the choroid/sclera combined mirrored these directional changes. Feeding RA (25 mg/kg) repeatedly to guinea pigs, also resulted in rapid eye elongation (up to 5 times normal), and yet the RE was not effected. In conclusion, RA may act as a signal for the direction of ocular growth. PMID- 14751550 TI - Age-dependent decrease of retinal kynurenate and kynurenine aminotransferases in DBA/2J mice, a model of ocular hypertension. AB - The study examines age-dependent changes of kynurenic acid (KYNA) content and kynurenine aminotransferases (KAT I and KAT II) celluar expression in the retinas of DBA/2J mice. Retinas were obtained from DBA/2J mice of different ages (3, 6 and 11 months). C57BL6 mice were used as controls. As measured with HPLC, KYNA content decreased (p < 0.01) in the retinas of 6-month-old DBA/2J mice and continued to decrease (p < 0.0074) in the retinas of 11-month-old animals compared to the controls. Immunohistochemistry showed that expression of both KAT I and KAT II decreased markedly in the retinas of 11-month-old DBA/2J mice compared to controls. The impairment in KYNA biosynthesis in the retinas of DBA/2J mice may be one of the mechanisms of retinal neurodegeneration related to ocular hypertension. PMID- 14751551 TI - Monocular and dichoptic masking effects on the frequency doubling illusion. AB - The purpose of this paper was to investigate the effects of both monocular and dichoptic masking on the frequency doubling (FD) illusion, using both temporal and spatial masks. Monocular spatial tuning effects occurred around the fundamental FD spatial frequency of 0.25 cycles per degree (c/deg), whereas dichoptic spatial frequency tuning effects occurred at the doubled spatial frequency of 0.50 c/deg. Temporal tuning effects were observed at the FD temporal frequency of 25 Hz, in both monocular and dichoptic paradigms. These results suggest that the FD illusion is cortical in origin and is dominated by a flicker component. PMID- 14751552 TI - BOLD fMRI and psychophysical measurements of contrast response to broadband images. AB - We have measured the relationship between image contrast, perceived contrast, and BOLD fMRI activity in human early visual areas, for natural, whitened, pink noise, and white noise images. As root-mean-square contrast increases, BOLD response to natural images is stronger and saturates more rapidly than response to the whitened images. Perceived contrast and BOLD fMRI responses are higher for pink noise than for white noise patterns, by the same ratio as between natural and whitened images. Spatial phase structure has no measurable effect on perceived contrast or BOLD fMRI response. The fMRI and perceived contrast response results can be described by models of spatial frequency response in V1, that match the contrast sensitivity function at low contrasts, and have more uniform spatial frequency response at high contrasts. PMID- 14751553 TI - Depth-dependent contrast gain-control. AB - Contrast adaptation that was limited to a small region of the peripheral retina was induced as observers viewed a multiple depth-plane textured surface. The small region undergoing contrast adaptation was present only in one depth-plane to determine whether contrast gain-control is depth-dependent. After adaptation, observers performed a contrast-matching task in both the adapted and a non adapted depth-plane to measure the magnitude and spatial specificity of contrast adaptation. Results indicated that contrast adaptation was depth-dependent under full-cue (disparity, linear perspective, texture gradient) conditions; there was a highly significant change in contrast gain in the depth-plane of adaptation and no significant gain change in the unadapted depth-plane. A second experiment showed that under some monocular viewing conditions a similar change in contrast gain was present in the adapted depth-plane despite the absence of disparity information for depth. Two control experiments with no-depth displays showed that contrast adaptation can also be texture- and location-dependent, but the magnitude of these effects was significantly smaller than the depth-dependent effect. These results demonstrate that mechanisms of contrast adaptation are conditioned by 3-D and 2-D viewing contexts. PMID- 14751554 TI - Letter-recognition and reading speed in peripheral vision benefit from perceptual learning. AB - Visual-span profiles are plots of letter-recognition accuracy as a function of letter position left or right of the midline. Previously, we have shown that contraction of these profiles in peripheral vision can account for slow reading speed in peripheral vision. In this study, we asked two questions: (1) can we modify visual-span profiles through training on letter-recognition, and if so, (2) are these changes accompanied by changes in reading speed? Eighteen normally sighted observers were randomly assigned to one of three groups: training at 10 degrees in the upper visual field, training at 10 degrees in the lower visual field and a no-training control group. We compared observers' characteristics of reading (maximum reading speed and critical print size) and visual-span profiles (peak amplitude and bits of information transmitted) before and after training, and at trained and untrained retinal locations (10 degrees upper and lower visual fields). Reading speeds were measured for six print sizes at each retinal location, using the rapid serial visual presentation paradigm. Visual-span profiles were measured using a trigram letter-recognition task, for a letter size equivalent to 1.4x the critical print size for reading. Training consisted of the repeated measurement of 20 visual-span profiles (over four consecutive days) in either the upper or lower visual field. We also tracked the changes in performance in a sub-group of observers for up to three months following training. We found that the visual-span profiles can be expanded (bits of information transmitted increased by 6 bits) through training with a letter recognition task, and that there is an accompanying increase (41%) in the maximum reading speed. These improvements transferred, to a large extent, from the trained to an untrained retinal location, and were retained, to a large extent, for at least three months following training. Our results are consistent with the view that the visual span is a bottleneck on reading speed, but a bottleneck that can be increased with practice. PMID- 14751555 TI - Spontaneous eye movements in goldfish: oculomotor integrator performance, plasticity, and dependence on visual feedback. AB - To quantify performance of the goldfish oculomotor neural integrator and determine its dependence on visual feedback, we measured the relationship between eye drift-velocity and position during spontaneous gaze fixations in the light and in the dark. In the light, drift-velocities were typically less than 1 deg/s, similar to those observed in humans. During brief periods in darkness, drift velocities were only slightly larger, but showed greater variance. One hour in darkness degraded fixation-holding performance. These findings suggest that while visual feedback is not essential for online fixation stability, it may be used to tune the mechanism of persistent neural activity in the oculomotor integrator. PMID- 14751556 TI - Uncertainty in visual processes predicts geometrical optical illusions. AB - It is proposed in this paper that many geometrical optical illusions, as well as illusory patterns due to motion signals in line drawings, are due to the statistics of visual computations. The interpretation of image patterns is preceded by a step where image features such as lines, intersections of lines, or local image movement must be derived. However, there are many sources of noise or uncertainty in the formation and processing of images, and they cause problems in the estimation of these features; in particular, they cause bias. As a result, the locations of features are perceived erroneously and the appearance of the patterns is altered. The bias occurs with any visual processing of line features; under average conditions it is not large enough to be noticeable, but illusory patterns are such that the bias is highly pronounced. Thus, the broader message of this paper is that there is a general uncertainty principle which governs the workings of vision systems, and optical illusions are an artifact of this principle. PMID- 14751558 TI - The expanding role of PI3-kinase in bone. AB - Phosphatidylinositol-3-kinases (PI3-Ks) play an important role in signal transduction and have been implicated in mediating a broad range of cellular responses. There are three classes of PI3-Ks [I (a and b subclasses), II, and III] with different substrate specificities and different modes of regulation. In osteoclasts, PI3-K has been shown to be a critical downstream effector from at least three cell-surface receptors, c-fms [the receptor for colony-stimulating factor 1 (CSF-1)], alphaVB3 integrin, and RANK [receptor activator of nuclear factor-kB (NF-kB)]. Furthermore, PI3-K is known to partner with the cytoplasmic tyrosine kinase c-src in mediating the effects of activated c-fms. The effector actions of PI3-K are diverse, including influencing osteoclast survival and activity, mediating actin remodeling and motility, and regulation of attachment structures. Less is known about the roles of PI3-K in osteoblasts. However, recent evidence suggests a role for PI3-K in osteoblast differentiation and survival. The classification, structure, function, and regulation of PI3-Ks will be reviewed here, with particular emphasis on the role of PI3-K in bone. PMID- 14751559 TI - Signalling by fibroblast growth factor receptor 3 and parathyroid hormone-related peptide coordinate cartilage and bone development. AB - Bone development is regulated by conserved signalling pathways that are linked to multifunctional growth factors and their high affinity receptors. Parathyroid hormone-related peptide (PTHrP) and fibroblast growth factor receptor 3 (FGFR3) have been shown to play pivotal, and sometimes complementary, roles in the replication, maturation and death of chondrocytes during endochondral bone formation. To gain further insight into how these pathways coordinate cartilage and bone development, we generated mice lacking expression of both PTHrP and FGFR3. The phenotype of compound mutant mice resembled that of their PTHrP deficient littermates with respect to neonatal lethality, facial dysmorphism and foreshortening of the limbs. The absence of PTHrP in the developing epiphyseal cartilage of PTHrP-/- and PTHrP-/-/FGFR3-/- mice resulted in a dominant hypo proliferative phenotype. However, abnormalities such as the presence of nonhypertrophic cells among hypertrophic chondrocytes and excessive apoptosis seen in the hypertrophic zone of PTHrP-/- mice were absent in the PTHrP-/-/FGFR3 /- mice. Furthermore, the absence of FGFR3 in single and compound mutant mice led to decreased expression of vascular endothelial growth factor (VEGF) and an increase in depth of hypertrophic chondrocytes. These observations indicate that FGFR3 deficiency can rescue some of the defects seen in PTHrP-deficient mice and that it plays an important role in the regulation of chondrocyte differentiation and hypertrophy. These studies support a dominant role for PTHrP in regulating the pool of proliferating cells during limb development and suggest that signalling by FGFR3 plays a more prominent role in cartilage maturation and vascular invasion at the chondro-osseous junction. PMID- 14751560 TI - Overexpression of FGFR3, Stat1, Stat5 and p21Cip1 correlates with phenotypic severity and defective chondrocyte differentiation in FGFR3-related chondrodysplasias. AB - Achondroplasia (ACH) and thanatophoric dysplasia (TD) are human skeletal disorders of increasing severity accounted for by mutations in the fibroblast growth factor receptor 3 (FGFR3). Attempts to elucidate the molecular signaling pathways leading to these phenotypes through mouse model engineering have provided relevant information mostly in the postnatal period. The availability of a large series of human fetuses including 14 ACH and 26 TD enabled the consequences of FGFR3 mutations on endogenous receptor expression during the prenatal period to be assessed by analysis of primary cultured chondrocytes and cartilage growth plates. Overexpression and ligand-independent phosphorylation of the fully glycosylated isoform of FGFR3 were observed in ACH and TD cells. Immunohistochemical analysis of fetal growth plates showed a phenotype-related reduction of the collagen type X-positive hypertrophic zone. Abnormally high amounts of Stat1, Stat5 and p21Cip1 proteins were found in prehypertrophic hypertrophic chondrocytes, the extent of overexpression being directly related to the severity of the disease. Double immunostaining procedures revealed an overlap of FGFR3 and Stat1 expression in the prehypertrophic-hypertrophic zone, suggesting that constitutive activation of the receptor accounts for Stat overexpression. By contrast, expression of Stat and p21Cip1 proteins in the proliferative zone differed only slightly from control cartilage and differences were restricted to the last arrays of proliferative cells. Our results indicate that FGFR3 mutations in the prenatal period upregulate FGFR3 and Stat-p21Cip1 expression, thus inducing premature exit of proliferative cells from the cell cycle and their differentiation into prehypertrophic chondrocytes. We conclude that defective differentiation of chondrocytes is the main cause of longitudinal bone growth retardation in FGFR3-related human chondrodysplasias. PMID- 14751561 TI - Matrix metalloproteinase-12 (MMP-12) in osteoclasts: new lesson on the involvement of MMPs in bone resorption. AB - Osteoclasts require matrix metalloproteinase (MMP) activity and cathepsin K to resorb bone, but the critical MMP has not been identified. Osteoclasts express MMP-9 and MMP-14, which do not appear limiting for resorption, and the expression of additional MMPs is not clear. MMP-12, also called metalloelastase, is reported only in a few cells, including tissue macrophages and hypertrophic chondrocytes. MMP-12 is critical for invasion and destruction in pathologies such as aneurysm and emphysema. In the present study, we demonstrate that osteoclasts express MMP 12, although only in some situations. Northern blots show that highly purified rabbit osteoclasts in culture express MMP-12 at the same level as macrophages, whereas in situ hybridizations performed on rabbit bone do not show any MMP-12 expression in osteoclasts whatever the bone type. In contrast, in situ hybridizations performed on mouse bone show MMP-12 expression in osteoclasts in calvariae and long bones. We also demonstrate that recombinant MMP-12 cleaves the putative functional domains of osteopontin and bone sialoprotein, two bone matrix proteins that strongly influence osteoclast activities, such as attachment, spreading and resorption. Furthermore, we investigated the role of MMP-12 in bone resorption and osteoclast recruitment by comparing MMP-12 knockout and wild-type mice in specialized culture models known to depend on MMP activity, as well as in the ovariectomy model, and we did not find any indication for a limiting role of MMP-12 in these processes. In conclusion, we found that osteoclasts are able to express MMP-12, but MMP-12 did not appear critical for osteoclast recruitment or resorption. The fact that none of the MMPs identified so far in osteoclasts appears limiting for resorption, gives strength to the hypothesis that the critical MMP for bone solubilization is produced by non-osteoclastic cells. PMID- 14751562 TI - Immunolocalization of matrix metalloproteinase-13 on bone surface under osteoclasts in rat tibia. AB - Matrix metalloproteinase (MMP)-13 (an interstitial collagenase also called collagenase 3) is involved in degradation of extracellular matrix in various tissues. Using immunohistochemistry and Western blotting, we investigated localization of MMP-13 in rat tibia, to clarify the role of MMP-13 in bone resorption. MMP-13 reactivity was mainly seen on bone surfaces under osteoclasts, and in some osteocytes and their lacunae near osteoclasts. However, immunoreactivity was not seen in chondrocytes or osteoclasts. MMP-13 was also localized on cement lines in the epiphysis. In the growth plate erosion zone, perivascular cells showed MMP-13 reactivity. Immunoelectron microscopy revealed that MMP-13 was localized on the bone surfaces, under the ruffled borders and some clear zones of osteoclasts. Gold-labeled MMP-13 was closely associated with collagen fibrils. Gold labeling was also detected in Golgi apparatus of osteocytes adjacent to osteoclasts and bone lining cells. Western blotting showed that MMP-13 was mainly associated with mineralized bone matrix. These findings suggest that MMP-13 synthesized and secreted by osteoblast-lineage cells is localized under the ruffled borders of osteoclasts. MMP-13 may play an important role in degradation of type I collagen in bone matrix, acting in concert with cathepsin K and MMP-9 produced by osteoclasts. MMP-13 in perivascular cells may be involved in removal of cartilage matrix proteins such as type II collagen and aggrecan. PMID- 14751563 TI - Transforming growth factor-beta induces osteoclast formation in the absence of RANKL. AB - Transforming growth factor beta (TGFbeta) is a multifunctional growth factor that is produced by many cells in bone and is abundant in the bone matrix. TGFbeta is known to regulate RANKL-induced osteoclast formation and bone resorbing activity. In this study we sought to determine whether TGFbeta could directly induce osteoclast formation by a RANKL-independent mechanism. We found that the addition of TGFbeta to cultures of human monocytes and RAW 264.7 cells (in the presence of M-CSF and the absence of RANKL, TNFalpha or IL-6/IL-11) was sufficient to induce the formation of TRAP+ and VNR+ cells, which formed actin rings and were capable of extensive lacunar resorption. The addition of osteoprotegerin or antibodies to TNFalpha and its receptors, as well as antibodies to gp130, did not inhibit lacunar resorption, indicating that TGFbeta did not act by stimulating RANKL, TNF or IL-6 production by monocytes. TGFbeta-induced osteoclast formation was qualitatively different from that induced by RANKL with numerous TRAP+/VNR+ mononuclear and small multinucleated cells being formed; these cells produced many small resorption lacunae. Our results indicate that TGFbeta, which is abundant in the bone matrix, can, in the presence of M-CSF, directly induce mononuclear phagocyte osteoclast precursors to differentiate into osteoclastic cells capable of lacunar resorption. PMID- 14751565 TI - Bone formation in the presence of platelet-rich plasma vs. bone morphogenetic protein-7. AB - Growth factors contained in platelet-rich plasma (PRP) have recently been proposed to enhance maturation of bone grafts and, in combination with anorganic bovine bone, to support repair in the treatment of small bone defects in maxillofacial surgery. Bone morphogenetic proteins (BMP) carried in a matrix may be able to replace the autologous bone graft in the treatment of critical size defects. However, no studies have compared the bone stimulating capacity of PRP and BMP. Likewise there is no data comparing the effects of PRP in either an autologous bone graft or in anorganic bovine bone. We augmented the mandible of Wistar rats (n = 28) on both sides with either anorganic bovine bone (Bio-Oss) or autologous rib bone. On the test side we applied either 20 microl of autologous PRP or 10 microl of rhBMP-7 (4 groups, n = 7). In addition, bone induction was evaluated in an extraskeletal site (n = 14). A polychrome sequential labeling was performed. The animals were sacrificed by intra-vital perfusion on day 50. Undecalcified ground sections were evaluated by microradiography, digitized histomorphometry and under fluorescent light. The qualitative analysis of fluorochrome labels suggested that PRP and rhBMP-7 accelerated bone growth. However, histomorphometric analysis revealed no significant differences in the area of newly mineralized bone under either the influence of PRP or rhBMP-7 on autologous bone graft. Likewise, the addition of PRP to anorganic bovine bone showed no statistical difference to the control group. The strongest bone stimulating effect was seen for the combination of rhBMP-7 with anorganic bovine bone (p = 0.028). In the extraskeletal model, newly formed bone was evident in the presence of rhBMP-7, but not of PRP. In conclusion, according to the histomorphometry, the addition of platelet-rich plasma failed to enhance bone formation on anorganic bovine bone and on autologous bone grafts. PMID- 14751566 TI - Histamine participates in the early phase of trabecular bone loss in ovariectomized rats. AB - We have previously reported that cimetidine, a reference H2 receptor antagonist, attenuates the initial osteoclastic burst and subsequent trabecular bone loss induced by ovariectomy (ovx) in rats. This study was designed to determine whether these effects are specific to H2 antagonism. To this end, we compared the effects of two H2 receptor antagonists, cimetidine and famotidine. In addition, we analyzed the response of histamine-producing cells to these inhibitors. Seventy-two 90-day-old female Sprague-Dawley rats were ovariectomized or sham operated, and received single daily intramuscular injections of cimetidine (125 mg/kg), famotidine (10 mg/kg), or vehicle. The animals were killed 14 days after surgery and their femurs were processed for histomorphometry. Trabecular bone volume was reduced by 30% in ovx rats and by 15% in cimetidine- and famotidine treated rats. Architectural parameters were reduced by about 20% in ovx rats. Cimetidine and famotidine attenuated these consequences of ovx by about 50%. Trabecular connectivity was deteriorated by ovx, while cimetidine and famotidine attenuated this effect. Resorption parameters were increased by ovx, while cimetidine and famotidine prevented this increase. Kinetic bone formation parameters were increased by ovx, while cimetidine and famotidine had no influence. Neither cimetidine nor famotidine had any observable effect in sham treated rats. Mast cell numbers increased by 250% in ovx rats and by only 40% in H2 antagonists-treated ovx rats. A resident histamine-positive, non-mast cell, population found in bone marrow was increased by 25% by ovx. Interestingly, cimetidine and famotidine reduced this population in both sham-operated and ovx rats, famotidine being more potent than cimetidine. These results show that H(2) receptor blockade partially prevents the consequences of castration on cancellous bone resorption in female rats, and strongly suggest that histamine participates in the mediator network regulating estrogen deficiency induced bone resorption. A large population of histamine-producing cells, which differ morphologically from mast cells and belong to an immature marrow population, may be a source of histamine in this model. The H(2) blockers targeted this population, and this effect appeared to explain the anti-resorptive action of the two drugs. PMID- 14751564 TI - Reduction of c-Src activity by substituted 5,7-diphenyl-pyrrolo[2,3-d] pyrimidines induces osteoclast apoptosis in vivo and in vitro. Involvement of ERK1/2 pathway. AB - We employed potent and selective c-Src inhibitors to investigate the functional and molecular consequences of inhibited c-Src tyrosine kinase activity in osteoclasts. These pyrrolopyrimidine derivatives reduced osteoclast numbers and induced osteoclast disruption in vivo. In vitro, they inhibited resorption pit formation and osteoclastogenesis, impaired adhesion ability and actin ring organization, and induced programmed cell death in mature osteoclasts. The cell death receptor Fas and p53 were insensitive to c-Src modulation. The expression of the cyclin-dependent kinase (CDK)-inhibitor p21WAF1/CIP1 was markedly reduced, but neither Bcl-2 nor Bcl-xL or Bax were modulated by c-Src inhibition. Caspase 9, and to a lesser extent caspase-3, but not caspase-8, were transiently cleaved (activated) by treatment with the c-Src inhibitors. c-Src inhibition stabilized p38 mitogen-activated protein kinase (MAPK), whereas the c-Jun N-terminal kinase (JNK) pathway did not appear to be modulated by our compounds. Most interestingly, transient extracellular signal regulated kinase (ERK1/2) dephosphorylation followed by sustained remarkable rephosphorylation overwhelming control levels was observed in response to c-Src inhibition. Blockade of ERK1/2 rephosphorylation by PD98059 reduced osteoclast nuclear disruption, suggesting the involvement of this pathway in apoptosis. Collectively, these data demonstrate that small pyrrolopyrimidine derivatives impair osteoclast function and induce cell damage suggestive of apoptosis in vivo and in vitro, with mechanisms presumably involving selective sustained ERK1/2 phosphorylation. PMID- 14751567 TI - Interaction of estrogen receptor alpha with protein kinase C alpha and c-Src in osteoblasts during differentiation. AB - In cultured osteoblasts, protein kinase C (PKC) activity increases and estrogen receptor alpha (ERalpha) binding capacity decreases upon confluence. We investigated potential interactions between ERalpha and PKC isoforms and their confluence-induced modulations in clonal ROS.SMER#14 cells and primary osteoblasts. In sub-confluent ROS.SMER#14 cells, which express an exogenous plus small amounts of the endogenous ERalpha gene, the receptor appeared as two main bands of approximately 66 and approximately 46 kDa. In over-confluent, more differentiated cells, the cytosolic approximately 66 kDa ERalpha appeared decreased and the approximately 46 kDa variant increased. Enhanced expression and/or membrane translocation of PKCalpha and PKCepsilon, but not PKCzeta, was evidenced at over-confluence, along with transient increases in expression and kinase activity of c-Src, accompanied by membrane translocation of the kinase activated enzyme. In contrast, negligible membrane translocation of PKCalpha and/or activated c-Src was observed in parental ROS 17/2.8 cells, which express low levels of full-length ERalpha. PKCalpha from over-confluent cells phosphorylated p60c-Src in vitro, suggesting functional interaction between the two kinases. ERalpha co-immunoprecipitated c-Src and PKCalpha, mostly in its cleaved form (PKMalpha). An analogous interaction was observed in primary osteoblasts. However, in these cells, much more PKCalpha/PKMalpha was ERalpha-co immunoprecipitated at over-confluence, a condition in which the shorter, approximately 46 kDa ERalpha variant is increased. This interaction was enhanced by estradiol treatment or PKC down-regulation, but was unaffected by c-Src inhibition. These data highlight direct PKCalpha-c-Src-ERalpha interactions, which may be crucial in the modulation of estrogen responsiveness and the differentiation process in osteoblasts. PMID- 14751568 TI - The nitrogen-containing bisphosphonate, zoledronic acid, increases mineralisation of human bone-derived cells in vitro. AB - Previous studies have attributed the increase in bone mass observed following bisphophonate (BP) therapy to their effects on bone-resorbing osteoclasts (OCs). However, recent evidence suggests that BPs can also act directly on bone forming osteoblasts (OBs) to increase their anabolic activity. Using an established model of in vitro OB differentiation, we found that the potent nitrogen-containing BP, zoledronic acid (ZOL), may enhance the bone forming potential of human adult OB like cells in vitro by inducing their differentiation. ZOL dose dependently induced both cytostasis and cell death in OB-like cells at concentrations of 0.5 microM or greater. Cells expressing high levels of the osteoprogenitor antigen, STRO-1, exhibited a greater proliferative potential than STRO-1negative/dim cells, and were more susceptible to the cytostatic and apoptotic effects of ZOL. ZOL was also found to promote bone cell differentiation, as evidenced by an increase in the number of cells exhibiting a more differentiated (STRO-1(-)/AP+ and STRO-1(-)/AP-) phenotype. Analysis of gene expression, using semi quantitative RT-PCR, demonstrated that ZOL treatment resulted in a significant upregulation of osteocalcin (OCN) and bone morphogenetic protein-2 (BMP-2) gene expression. Furthermore, in vitro mineralisation studies revealed that ZOL enhanced mineralised matrix formation at concentrations between 5 and 25 microM. These results show that, in addition to its direct effects on OCs, ZOL also directly affects the proliferation and differentiation of human OB-like cells in vitro and may enhance bone formation in vivo. PMID- 14751569 TI - mRNA expression and protein localization of dentin matrix protein 1 during dental root formation. AB - Dentin matrix protein 1 (DMP1) is an acidic phosphoprotein. DMP1 was initially detected in dentin and later in other mineralized tissues including cementum and bone, but the DMP1 expression pattern in tooth is still controversial. To determine the precise localization of DMP1 messenger RNA (mRNA) and the protein in the tooth, we performed in situ hybridization and immunohistochemical analyses using rat molars and incisors during various stages of root formation. During root dentin formation of molars, DMP1 mRNA was detected in root odontoblasts in parallel with mineralization of the dentin. However, the level of DMP1 mRNA expression in root odontoblasts decreased near the coronal part and was absent in coronal odontoblasts. DMP1 protein was localized along dentinal tubules and their branches in mineralized root dentin, and the distribution of DMP1 shifted from the end of dentinal tubules to the base of the tubules as dentin formation progressed. During the formation of the acellular cementum, DMP1 mRNA was detected in cementoblasts lining the acellular cementum where its protein was localized. During the formation of the cellular cementum, DMP1 mRNA was detected in cementocytes embedded in the cellular cementum but not in cementoblasts, and its protein was localized in the pericellular cementum of cementocytes including their processes. During dentin formation of incisors, DMP1 mRNA was detected in odontoblasts on the cementum-related dentin, where its protein was localized along dentinal tubules near the mineralization front. The localization of DMP1 mRNA and protein in dentin and cementum was related to their mineralization, suggesting that one of the functions of DMP1 may be involved in the mineralization of dentin and cementum during root formation. PMID- 14751570 TI - Effect of gene dose and parental origin on bone histomorphometry in X-linked Hyp mice. AB - X-linked hypophosphatemia (XLH) is characterized by rickets and osteomalacia and arises from mutations in the Phex and PHEX genes in mice (Hyp) and humans, respectively. The present study was undertaken to examine the effect of gene dose on the skeletal phenotype using a histomorphometric approach. Metrical traits (vertebral length, growth plate thickness, cancellous osteoid volume per bone volume, and cancellous, endocortical, and periosteal osteoid thickness) were compared in caudal vertebrae of mutant female (Hyp/+, Hyp/Hyp) and male (Hyp/Y) mice and their normal female (+/+) and male (+/Y) littermates. Mutant animals had trait values that differed significantly from those of normal animals. However, with the exception of vertebral length and cancellous osteoid thickness, values were not significantly different between the three mutant genotypes. We also examined the effect of gamete-of-origin on histomorphometric parameters in obligate Hyp/+ females derived from male or female transmitting parents. The metrical trait values in both groups of Hyp/+ mice were similar, with the exception of vertebral length and cancellous osteoid volume per bone volume. In summary, we demonstrate that the amount of osteoid per bone volume is similar in the three mutant genotypes and conclude that the extent and magnitude of the mineralization defect is fully dominant and likely not affected by gene dose. The differences in vertebral length in the mutants suggest that rickets and osteomalacia are not the only causes of decreased vertebral growth in Hyp mice and that Phex protein may influence bone growth and mineralization by distinct pathways. PMID- 14751571 TI - Reduction of nocturnal rise in bone resorption by subcutaneous GLP-2. AB - We have previously shown that a subcutaneous injection of glucagon-like peptide-2 (GLP-2) at 9 a.m. in fasting postmenopausal women results in a dose-dependent decrease in the serum concentration of fragments derived from the degradation of the C-terminal telopeptide region of collagen type I (s-CTX), a marker of bone resorption. In contrast, GLP-2 was found to have a neutral effect on bone formation, as assessed by serum osteocalcin. Since increased s-CTX levels are normally observed at night, we conducted bedtime studies in healthy postmenopausal women. The objective was to study the effect of GLP-2 injection on bone turnover given at bedtime. A total of 81 postmenopausal women were included in two randomised placebo-controlled studies. In conclusion, we found a dose related reduction of s-CTX after injection of GLP-2 (P < 0.05) and osteocalcin levels was increased as compared to placebo (P = 0.07) by the treatment, suggestive of a stimulative effect on bone formation. An area under the curve (AUC0-10 h) analysis for s-CTX after GLP-2 injection confirmed the dose-related decrease as compared to placebo (P < 0.05). PMID- 14751572 TI - Ionizing radiation sensitizes bone cells to apoptosis. AB - Osteoradionecrosis is a common sequelae of radiation therapy for head and neck cancer. To test the hypothesis that radiation induces osteoradionecrosis by induction of bone cell apoptosis, we exposed MC3T3-E1 osteoblast-like cells to gamma-radiation and evaluated cell viability. Twenty-four hours postirradiation, measurement of osteoblast dehydrogenase activity suggested that there was a small decrease in cell viability. However, TUNEL and flow cytometric analysis indicated that the viability loss was caused by inhibition of cell proliferation and not by induction of apoptosis. The effect of irradiation on osteoblast function was examined by Western blot and flow cytometric analysis. It was found that irradiated osteoblasts underwent G2 cell cycle arrest. In addition, we observed changes in expression of molecules that regulate the cell cycle. Thus, there was an increase in p53 transcription, a raised level of MDM2 dephosphorylation, and elevation in p21 and GADD153 protein levels. Since these proteins are concerned with the regulation of the cell cycle, the observed changes in expression would be expected to disturb cyclin activity and cause G2M arrest. The arrested cells displayed a dramatic increase in sensitivity to specific apoptogens. Thus, when irradiated, and then treated with Ca2+Pi or staurosporine, agents that cause mitochondrial dysfunction, more osteoblasts underwent apoptosis than with the apoptogen alone. In contrast, irradiated cells treated with anti-Fas antibody showed no change in apoptotic sensitivity; apoptosis was inhibited when osteoblasts were treated with etoposide. Similar alterations in sensitivity were observed when cells were arrested in G2/M by pretreatment with colchicine and then challenged with apoptogens. It was concluded that activation of radiation induced G2 arrest sensitizes osteoblasts to agents that mediate apoptosis through a mitochondrial-dependent death pathway. PMID- 14751573 TI - A selective EP4 receptor antagonist abrogates the stimulation of osteoblast recruitment from bone marrow stromal cells by prostaglandin E2 in vivo and in vitro. AB - Recent evidence indicates that systemic administration of PGE2 increases bone formation and bone mass via activation of the EP4 receptor. Previously, we demonstrated that osteoblastic recruitment from rat bone marrow stromal cells (BMSC) is a major mechanism for the anabolic effect of PGE2. In this study, we used a selective EP4 antagonist to test if the stimulation of osteoblast differentiation from rat BMSC in vitro and in vivo involves the EP4 receptor. In vitro, PGE2 (100 nM) increased nodule formation and alkaline phosphatase (ALP) activity in cultures of rat BMSC 1.5- to 2-fold. These effects were abolished by the EP4 antagonist at 10(-6) M but not 10(-9) M. Furthermore, PGE2 increased the number of surviving adherent BMSC by approximately 225% and the EP4 antagonist prevented this effect as well. The antagonist had no effect on basal levels of nodule formation and adherent cell number. In vivo, daily systemic administration of PGE2 at 6 mg/kg for 2 weeks increased cancellous bone area (by approximately 50%) and increased nodule formation (measured as mineralized area) in ex vivo stromal cultures by approximately 50%. Pre-administration of the EP4 antagonist at 10 mg/kg abrogated both the increase in bone mass as well as the increase in nodule formation. These data indicate that PGE2 stimulates osteoblastic commitment of BMSC via activation of the EP4 receptor. PMID- 14751574 TI - Detecting and tracking local changes in the tibiae of individual rats: a novel method to analyse longitudinal in vivo micro-CT data. AB - In this study we present the analysis of in vivo micro-CT scans using a new method based on image registration that accurately evaluates longitudinal micro CT studies. We tested if detailed changes in the bone architecture could be detected and tracked in individual animals. A prototype in vivo micro-CT scanner (Skyscan 1076) was developed in which tibiae of rats that are lying on a bed under gas anaesthesia were scanned. For this study, three female Wistar rats were used: a sham-operated rat, an ovariectomised (OVX) rat and one rat that served as a reproducibility control. The reproducibility control rat was scanned twice in 1 day. The other animals were scanned at week 0, just before surgery, at week 4 and at week 14 after surgery. Architectural changes over time were detected by overlaying two data sets made at different time points using an algorithm that uses mutual information for optimal registration. The scans were segmented into binary data sets using a local thresholding algorithm. The reproducibility test showed small errors of less than 3% in bone volume measurements and errors less than 0.5% in measurements of trabecular thickness. The sham-operated rat showed no changes in total bone volume, though thinning and eventual loss of some small trabeculae could be detected, which could be related to the age of the animal. The OVX rat lost much trabecular bone volume, especially in the metaphysis (60% at week 4, 75% at week 14). The remaining trabeculae slowly increased in thickness. Following the different scans in time showed the forming of new trabecular structures. Additionally, small longitudinal growth at the growth plate could be detected after the first 4 weeks. Further, the OVX rat showed extensive modelling at the proximal endosteal lateral cortex. We have shown a new method that can detect and track changes in the local bone architecture and individual trabeculae in time, in an individual living animal. This method enables longitudinal in vivo micro-CT studies and has the potential to greatly contribute to experimental rat or mouse studies on pharmacological intervention and transgenic models. PMID- 14751575 TI - Analysis of the influence of image resolution on the discriminating power of trabecular bone architectural parameters. AB - In the study the influence of image resolution on the discriminating power of different parameters quantifying the architecture of trabecular bone is investigated. High-resolution images of 200-microm-thick sections of young and old trabecular bone are collected. Different architectural parameters are evaluated and shown to be statistically different in both groups of images. Then the resolution of the images is artificially degraded to the level comparable with resolution achievable in vivo. It is shown that although the errors of evaluation are quite large for low-resolution images, the statistical difference present in the original data is still observed for parameters that depend only on the global characteristics of trabecular structure or marrow space, or depend only linearly on the number and area of disconnected parts of marrow space or trabecular structure. The parameters that fulfill such conditions are Euler number, mean area of marrow cavities, star volume and trabecular spacing. The statistical difference disappears for parameters that depend on the number and area of disconnected parts of marrow space or trabecular structure in a nonlinear manner, that is, marrow and trabecular disconnection probabilities, two-point distance and second moment of marrow cavities area distribution. It is shown that the mechanism that leads to the losing of discriminating power does not depend crucially on the estimation error or noise level. This mechanism, which involves an interplay between changes of mean values and standard deviations, is the manifestation of partial volume effect amplified with image segmentation. PMID- 14751576 TI - Body composition and bone measurements in intra-uterine growth retarded and early postnatally undernourished male and female rats at the age of 6 months: comparison with puberty. AB - Undernutrition in early life may permanently change body structure, physiology and metabolism and leads to chronic diseases in later life. To test whether malnutrition during different critical time periods around birth in the rat has long-lasting effects on body composition and skeletal growth, we examined body weight and body composition in pubertal rats and adult rats of 6 months after pre and postnatal malnutrition. Prenatal malnutrition or intra-uterine growth retardation (IUGR) was induced by ligation of the uterine arteries on day 17 of gestation and postnatal food restriction (FR) by litter enlargement to 20 pups per mother from day 2 after birth until weaning (day 24). Pubertal markers were balanopreputial separation (BPS) in the male and vaginal opening (VO) in the female. IUGR as well as FR resulted in a persistent growth retardation. From birth in IUGR rats and from day 4 after birth in FR rats until 6 months of age body weight in male and female rats was significantly lower compared with controls (P < 0.01 and P < 0.05). Although total body bone mineral content (TBBMC) did not differ between male IUGR rats and controls at BPS, at the age of 6 months TBBMC was significantly lower (P < 0.01) compared with controls. At BPS as well as at 6 months of age, TBBMC was significantly lower in male FR rats compared with controls (P < 0.05 and P < 0.01). In the female IUGR rats TBBMC was significantly lower compared with controls at VO (P < 0.01) and 6 months (P < 0.05). TBBMC in the female FR rats was significantly lower at VO (P < 0.01), but did not differ from controls at the age of 6 months. For both IUGR and FR male and female rats these differences disappeared after adjusting for body weight. Body composition in terms of total fat mass, percentage fat and percentage lean did not differ from controls in male and female IUGR rats at 6 months and the same results were observed in the female FR rats. However, in the male FR rats, total fat mass and percentage fat were significantly lower compared with controls (P < 0.01 and P < 0.05), while the percentage lean mass was significantly higher (P < 0.05). We conclude that different critical time periods of malnutrition around birth have different effects later in life on growth, which do not disappear at least after 6 months of life. With respect to body composition, only in the FR male rats, differences are found in total fat mass and the balance of percentage fat mass and lean mass. At time of puberty and at the age of 6 months bone mass adjusted for body weight does not seem to be affected by perinatal undernutrition. PMID- 14751577 TI - Clinical performance of immunoreactive tartrate-resistant acid phosphatase isoform 5b as a marker of bone resorption. AB - Previous immunoassays developed for the measurement of serum tartrate-resistant acid phosphatase (TRACP) have lacked specificity for osteoclastic TRACP, TRACP 5b, or have not shown satisfactory clinical performance. The aim of this study was to evaluate the clinical performance of a novel immunocapture activity assay for TRACP 5b, in comparison to telopeptide fragments of type I collagen. Within subject variability and the effect of feeding on TRACP 5b and telopeptides of type I collagen were assessed in 20 healthy premenopausal women. Diurnal variation of TRACP 5b and serum beta C-terminal cross-linked telopeptide of type I collagen (sbetaCTX) was assessed in 12 healthy postmenopausal women. Renal clearance was assessed in 19 end stage renal failure patients undergoing routine haemodialysis. Response to antiresorptive treatment and calcium supplementation was assessed in osteoporotic postmenopausal women treated with alendronate and calcium (n = 16) or with calcium alone (n = 7) for 24 weeks.Within-subject variability (CVi) of TRACP 5b was 6.6%, lower than CVi of urinary and serum telopeptides. TRACP 5b decreased by 2.4 +/- 0.8%, in response to feeding (P < 0.05) compared to 7.0 +/- 2.6% to 7.9 +/- 3.7% for urinary telopeptides (P < 0.05 to < 0.01) and 8.5 +/- 1.7% to 17.8 +/- 2.6% for serum telopeptides (P < 0.0001). The amplitude of the diurnal rhythm for TRACP 5b was small compared to that of sbetaCTX, 14 +/- 4% vs. 137 +/- 14%. Haemodialysis did not have a significant effect on TRACP 5b but reduced sbetaCTX by 46 +/- 4% (P < 0.0001). In response to alendronate, TRACP 5b decreased by 39 +/- 4% compared to 49 +/- 4% to 69 +/- 5% for urinary telopeptides and 75 +/- 8% for sbetaCTX. We conclude that TRACP 5b shows an attenuated response to antiresorptive therapy in comparison with other markers of bone resorption, but that this may be offset by lower biological variability. TRACP 5b may provide useful additional information about bone resorption. PMID- 14751578 TI - Fracture incidence and association with bone mineral density in elderly men and women: the Rotterdam Study. AB - The incidence of all non-vertebral fractures, as well as the relation to bone mineral density (BMD), was quantified in 7806 men and women from the Rotterdam Study, a prospective, population-based cohort study of men and women aged 55 years and older. In addition, the sensitivity of using a T-score at or below -2.5 for identifying subjects at risk for fractures was assessed. At baseline, between 1990 and 1993, femoral neck BMD was measured by dual energy X-ray absorptiometry (DXA). Subsequently, gender-specific T-scores were calculated using the NHANES reference population. During a mean follow-up of 6.8 years, information on incident non-vertebral fractures was gathered. In general, hip, wrist and upper humerus fractures are the most frequent fractures in both men and women. Femoral neck BMD appears to be an equally important risk factor in both genders, and is especially related to hip fractures. For all non-vertebral fractures, the age adjusted hazard ratio (95% confidence interval) per standard deviation decrease in femoral neck BMD was 1.5 (1.4-1.6) for women and 1.4 (1.2-1.6) for men. For hip fractures, the hazard ratios were 2.1 (1.7-2.5) for women and 2.3 (1.6-3.3) for men. Only 44% of all non-vertebral fractures occurred in women with a T-score below -2.5; in men, this percentage was even lower (21%). Thus, there is a clear need for the development of more sensitive risk assessment tools, using not only BMD, but also other clinical predictors of fractures. PMID- 14751579 TI - Effects of hypophysectomy and recombinant human growth hormone on material and geometric properties and the pre- and post-yield behavior of femurs in young rats. AB - To study the musculoskeletal effects of hypophysectomy (Hx) and a partial replacement treatment with recombinant human growth hormone (rhGH) in rats, we determined the stiffness (elastic modulus, E) and volumetric BMD (vBMD) of cortical bone; the periosteal and endosteal perimeters, area and bending moment of inertia (xCSMI) of the cross sections, and the structural stiffness and pre- and post-yield strength of the femur diaphyses by pQCT and mechanical tests, and the gastrocnemius weight of rats that were either intact (n = 9) or Hx at 15 days of age (20). The latter were otherwise untreated (Hx controls, 4) or given 0.4 (8) or 2.0 (8) IU kg(-1) day(-1), s.c., of rhGH for 45 days starting 15 days after surgery. Hx delayed musculoskeletal development (gastrocnemius weight, bone geometric properties), thus affecting the diaphyseal stiffness and strength. It also reduced the cortical vBMD through an undefined mechanism, and increased the elastic modulus of cortical bone. The Hx also affected the correlation between bone geometric and material properties (xCSMI vs. E), suggesting an antianabolic interaction with the biomechanical control of bone modeling in response to strains caused by mechanical usage. As a result, Hx reduced the stiffness, post yield, and ultimate strength of the diaphyses. These effects should reflect changes in bone tissue microstructure, perhaps associated with crack generation and progress, but unrelated to bone mineral mass. They are compatible with the induction of a delay in collagen turnover with associated increases in fibers' diameter and crystals' size that may have resulted from the suppression of some other hormones, such as thyroid, prolactin, or other hormones regulated by ACTH. The above doses of rhGH significantly but incompletely prevented the negative Hx effects on bone and muscle development (bone geometric properties, muscle mass). However, rhGH treatment failed to prevent the demineralizing and stiffening effect of Hx on bone tissue and the unusual effects on the post-yield strength (less clearly related to muscle development than the former). Consequently, rhGH treatment tended to preserve the natural relationship between muscle function and bone geometry but not bone strength. The effects of larger rhGH doses and the interaction of other hormones with the described effects remain to be investigated. Nevertheless, these findings would deserve special attention because they challenge the prevailing view that in endocrine-metabolic bone weakening diseases the bone matrix always has a normal composition. PMID- 14751580 TI - Association between fluoride, magnesium, aluminum and bone quality in renal osteodystrophy. AB - INTRODUCTION: Trace elements are known to influence bone metabolism; however, their effects may be exacerbated in renal failure because dialysis patients are unable to excrete excess elements properly. Our study correlated bone quality in dialysis patients with levels of bone fluoride, magnesium, and aluminum. A number of studies have linked trace elements, including fluoride, magnesium, and aluminum, to the development of renal osteodystrophy (ROD). However, little is known about the relationship between trace elements and changes in bone quality in ROD patients. The purpose of this study was to examine bone quality in ROD patients, and correlate differences in bone quality to trace element concentrations in bone. Bone quality encompasses parameters that contribute to the mechanical integrity of the bone. METHODS: One hundred fifty-three anterior iliac crest bone biopsies from patients with ROD were examined and subdivided into five groups based on the pathological features. Parameters contributing to bone quality, such as bone structure and remodeling, connectivity, mineralization, and microhardness, were assessed and correlated to bone chemical composition. In addition, clinical symptoms of ROD were assessed and correlated with bone composition. RESULTS AND CONCLUSIONS: There were no differences in bone architecture between the different ROD bone groups; however, differences in bone mineralization and microhardness were observed. Increase in bone fluoride was associated with increased osteoid parameters and decreased bone microhardness. Bone mineralization and microhardness decreased with increasing bone magnesium content and intact parathyroid hormone (PTH) level. Moreover, bone magnesium increased with intact PTH levels. The relationship between PTH, bone magnesium, mineralization, and microhardness was primarily observed in aplastic bone disorder. Furthermore, bone magnesium and aluminum contents were positively associated with bone pain and proximal myopathy in these patients. Most importantly, fluoride, magnesium, and aluminum showed significant correlations with one another. These results suggested that in ROD, bone fluoride may diminish bone microhardness by interfering with mineralization. Magnesium may be involved in the suppression of PTH secretion, lowering bone turnover thus leading to an increase in bone mineralization profile and microhardness in aplastic bone disorder. The effects of fluoride and magnesium on bone quality may be exacerbated by their interaction with aluminum. PMID- 14751581 TI - Shock wave treatment shows dose-dependent enhancement of bone mass and bone strength after fracture of the femur. AB - Shock wave treatment is believed to improve bone healing after fracture. The purpose of this study was to evaluate the effect of shock wave treatment on bone mass and bone strength after fracture of the femur in a rabbit model. A standardized closed fracture of the right femur was created with a three-point bending method in 24 New Zealand white rabbits. Animals were randomly divided into three groups: (1) control (no shock wave treatment), (2) low-energy (shock wave treatment at 0.18 mJ/mm2 energy flux density with 2000 impulses), and (3) high-energy (shock wave treatment at 0.47 mJ/mm2 energy flux density with 4000 impulses). Bone mass (bone mineral density (BMD), callus formation, ash and calcium contents) and bone strength (peak load, peak stress and modulus of elasticity) were assessed at 12 and 24 weeks after shock wave treatment. While the BMD values of the high-energy group were significantly higher than the control group (P = 0.021), the BMD values between the low-energy and control groups were not statistically significant (P = 0.358). The high-energy group showed significantly more callus formation (P < 0.001), higher ash content (P < 0.001) and calcium content (P = 0.003) than the control and low-energy groups. With regard to bone strength, the high-energy group showed significantly higher peak load (P = 0.012), peak stress (P = 0.015) and modulus of elasticity (P = 0.011) than the low-energy and control groups. Overall, the effect of shock wave treatment on bone mass and bone strength appears to be dose dependent in acute fracture healing in rabbits. PMID- 14751582 TI - On Jarvinen et al. (Bone 2003;32(6):642-61). PMID- 14751584 TI - Effects of lipopolysaccharide treatment on feeding of goldfish: role of appetite regulating peptides. AB - The gram-negative bacteria-derived endotoxin lipopolysaccharide (LPS) is known to play an important role in immune and neurological manifestations during bacterial infections. In mammals, peripheral or brain administration of LPS induces anorexia and is thought to exert its effects through activation of pro inflammatory cytokines. In this study, we investigated the effects of peripheral (intraperitoneal, IP) and central (intracerebroventricular, ICV) injections of LPS on food intake of goldfish. Fish treated IP with 10, 25, 50, 100 or 250 ng/g LPS or ICV with 1, 10 and 100 ng/g LPS showed a significant dose-dependent decrease in food intake, compared to the saline-treated fish. We also examined the brain mRNA expression of several hypothalamic appetite-related neuropeptides in response to the administration of LPS. IP injections of LPS (100 ng/g) induced a decrease in NPY expression and an increase in CCK, CRF and CART expression. These results indicate that LPS is a potent anorexigenic factor in goldfish and that this endotoxin induces a reduction in appetite, at least in part, by influencing gene expression of appetite-related neuropeptides. PMID- 14751585 TI - The cerebellar GABAA alpha6 subunit is differentially modulated by chronic ethanol exposure in normal (R100R) and mutated (Q100Q) sNP rats. AB - Sardinian alcohol non-preferring (sNP) rats carry a point mutation (R100Q) in the cerebellar expressed GABAA receptor alpha6 subunit gene, leading to a higher sensitivity to ethanol and diazepam. The role of the alpha6 subunit gene cluster in the ethanol non-preferring phenotype was here investigated by measuring the levels of alpha1, alpha6 and gamma2 peptide in the cerebellum of normal (RR) and mutated (QQ) sNP rats after 2 weeks of chronic ethanol administration. Western blot analysis revealed that the alpha6 subunit is increased in RR sNP rats after chronic ethanol exposure (25.44%+/-8.69 versus control), while it remained unchanged in mutated QQ sNP rats. Interestingly, chronic ethanol administration decreased alpha1 peptide levels in the cerebellum of both rat lines to a similar extent (30.99%+/-6.74 and 27.12%+/-9.83 in RR and QQ rats, respectively), while gamma2 peptide levels remained unchanged. To further correlate the genetic and biochemical difference of the normal and mutated sNP rats with their aversive phenotype, we exposed sNP rats to a protocol of acquisition and maintenance of ethanol drinking. QQ sNP rats drank less ethanol than RR rats during the acquisition phase, but such difference was lost during the maintenance phase. These data may contribute to elucidating the mechanisms of alcohol avoidance in rat lines selected for this behavior when exposed to ethanol solution. PMID- 14751587 TI - Age-related changes in growth hormone (GH) cells in the pituitary gland of male mice are mediated by GH-releasing hormone but not by somatostatin in the hypothalamus. AB - Using immunocytochemical and morphometric methods, we examine changes with age of growth hormone-releasing hormone (GHRH) in the arcuate nucleus (ARC), changes of somatostatin (SS) in the periventricular nucleus (PeN) of the hypothalamus, and changes of growth hormone (GH) cells in the anterior pituitary in male C57BL/6J mice at 2 months old (2 M), 4 M, 12 M and 24 M. The number of GHRH-ir neurons decreased significantly with age. The number of SS-ir neurons did not differ significantly between these all age groups. The volume of the anterior pituitary and the number of adenohypophysial parenchymal cells fell dramatically from 4 to 12 M. The proportion of GH-ir cells decreased significantly with age, and in absolute number from 4 to 12 M and in size from 2 to 4 M and from 4 to 12 M. These results suggest that the reduction in GH-ir cells in male mice is modulated by the reduction in GHRH-ir neurons, but not by SS-ir neurons. PMID- 14751586 TI - Homeostatic plasticity in hippocampal slice cultures involves changes in voltage gated Na+ channel expression. AB - Neurons preserve stable electrophysiological properties despite ongoing changes in morphology and connectivity throughout their lifetime. This dynamic compensatory adjustment, termed 'homeostatic plasticity', may be a fundamental means by which the brain normalizes its excitability, and is possibly altered in disease states such as epilepsy. Despite this significance, the cellular mechanisms of homeostatic plasticity are incompletely understood. Using field potential analyses, we observed a compensatory enhancement of neural excitability after 48 h of activity deprivation via tetrodotoxin (TTX) in hippocampal slice cultures. Because activity deprivation can enhance voltage-gated sodium channel (VGSC) currents, we used Western blot analyses to probe for these channels in control and activity-deprived slice cultures. A significant upregulation of VGSCs expression was evident after activity deprivation. Furthermore, immunohistochemistry revealed this upregulation to occur along primarily pyramidal cell dendrites. Western blot analyses of cultures after 1 day of recovery from activity deprivation showed that VGSC levels returned to control levels, indicating that multiple molecular mechanisms contribute to enhanced excitability. Because of their longevity and in vivo-like cytoarchitecture, we conclude that slice cultures may be highly useful for investigating homeostatic plasticity. Furthermore, we demonstrate that enhanced excitability involves changes in channel expression with a targeted localization likely profound transform the integrative capacities of hippocampal pyramidal cells and their dendrites. PMID- 14751588 TI - Differential expression of NMDA and AMPA receptor subunits in DARPP-32-containing neurons of the cerebral cortex, hippocampus and neostriatum of rats. AB - Dopamine and cyclic adenosine 3',5'-monophosphate-regulated phosphoprotein, 32 kDa (DARPP-32) is a key element of dopamine/D1/DARPP-32/protein phosphatase-1 (PP 1) signaling cascades of mammalian brain. We are interested in the expression patterns of N-methyl-D-aspartate (NMDA) and alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionate (AMPA) receptors in DARPP-32-containing neurons, which may constitute morphological basis for interaction between dopamine and ionotropic glutamate receptors in dopaminoceptive cells. Double immunofluorescence was performed to visualize neurons showing coexpression of DARPP-32 with NMDA or AMPA receptor subunits (i.e., NR1, NR2a/b, glutamate receptor subunit 1 [GluR1], GluR2/3, and GluR4) in the forebrains of rats. Distribution of DARPP-32-positive neurons completely or partially overlapped with that of NMDA receptor- or AMPA receptor-immunoreactive ones in the frontal and parietal cortex, hippocampus and neostriatum, and neurons double-labeled with DARPP-32/NR1, DARPP-32/NR2a/b, DARPP 32/GluR1, DARPP-32/GluR2/3, or DARPP-32/GluR4 immunoreactivity were numerously observed. Semiquantification analysis indicated that most of DARPP-32-containing neurons (86-98%) expressed NR1, NR2a/b and GluR2/3, while less of them (14-90%) expressed GluR1 and GluR4. Although high rates (90-98%) of DARPP-32-positive cells expressed NMDA receptors in all regions above, variant percentages of them expressing AMPA receptor subunits were observed among the cortex (54-90%), hippocampus (59-97%) and neostriatum (14-97%). The study presents differential expression patterns of NMDA and AMPA receptors in DARPP-32-postive neurons in these forebrain regions. Taken together with previous reports, the present data suggest that interaction between dopamine and glutamate receptors may occur in the dopaminoceptive neurons with distinct receptor compositions and may be involved in modulating neuronal properties and excitotoxicity in mammalian forebrain. PMID- 14751589 TI - Excitotoxic lesions of the prefrontal cortex attenuate the potentiation of amphetamine-induced locomotion by repeated neurotensin receptor activation. AB - This study was aimed at determining the role of prefrontal cortex neurons in the development of the potentiation of amphetamine-induced locomotor activity by repeated central injections of D-Tyr[11]neurotensin. Excitotoxic lesions of the prefrontal cortex were made by injecting bilaterally at three anterior-posterior placements 2 microg/microl of ibotenic acid. Ten days after surgery, locomotor responses to an intracerebroventricular injection of 0.18 or 18 nmol/10 microl of D-Tyr[11]neurotensin, or vehicle-saline, were measured in different groups of lesioned and sham rats. Ambulatory, non-ambulatory and vertical movements were measured for 2 h in activity cages starting immediately after the injection. This training phase was repeated on four occasions, every second day. One week after the last day of the training phase (day 14), locomotor responses to a single injection of amphetamine (0.75 mg/kg, IP) were measured in all rats. Results show that D-Tyr[11]neurotensin produced in sham animals a dose-dependent initial suppression of locomotor activity followed by an augmentation. The latter behavioral effect tended to be smaller in the lesioned rats, but not statistically different than in sham, suggesting that prefrontal cortex neurons do not play a major role in the stimulant effect of neurotensin on locomotor activity. However, sham rats pre-exposed to the high dose of D-Tyr[11]neurotensin showed stronger non-ambulatory and vertical movements than saline pre-exposed rats when tested with amphetamine; this sensitization effect was not observed in lesioned rats. The present results show that prefrontal cortex neurons are part of the neural circuitry involved in the development of amphetamine sensitization by repeated activation of central neurotensin receptors. PMID- 14751590 TI - Spatial frequency-dependent feedback of visual cortical area 21a modulating functional orientation column maps in areas 17 and 18 of the cat. AB - The feedback effect of activity of area 21a on orientation maps of areas 17 and 18 was investigated in cats using intrinsic signal optical imaging. A spatial frequency-dependent decrease in response amplitude of orientation maps to grating stimuli was observed in areas 17 and 18 when area 21a was inactivated by local injection of GABA, or by a lesion induced by liquid nitrogen freezing. The decrease in response amplitude of orientation maps of areas 17 and 18 after the area 21a inactivation paralleled the normal response without the inactivation. Application in area 21a of bicuculline, a GABAa receptor antagonist caused an increase in response amplitude of orientation maps of area 17. The results indicate a positive feedback from high-order visual cortical area 21a to lower order areas underlying a spatial frequency-dependent mechanism. PMID- 14751591 TI - Microdialysis study of striatal dopaminergic dysfunctions induced by 3 MPa of nitrogen- and helium-oxygen breathing mixtures in freely moving rats. AB - Previous studies have demonstrated opposite effects of high-pressure helium and nitrogen on extracellular dopamine (DA) levels, which may reflect disturbances on the synthesis, release or metabolic mechanisms. Intrastriatal microdialysis was used to measure the precursor (tyrosine), DA and its metabolites (DOPAC, HVA) levels under nitrogen- or helium- at pressure up to 3 MPa. Under 3 MPa of helium oxygen breathing mixtures, the extracellular concentration of tyrosine is decreased while the extracellular concentration of DA is increased. On the contrary, nitrogen-oxygen breathing mixture at the same pressure increased extracellular tyrosine concentration and decreased DA release. Under both conditions, an increment of the DOPAC and HVA levels could be noted. Our results suggest that changes in DA release and metabolism during high-pressure helium exposure reflect the effect of the pressure per se, whereas the intrinsic effects of narcotic gases, although sensitive to pressure, would be revealed by hyperbaric nitrogen exposure. PMID- 14751592 TI - Effect of the adenosine A2A receptor antagonist 8-(3-chlorostyryl)caffeine on L DOPA biotransformation in rat striatum. AB - In the present study, we investigated effects of the new selective adenosine A2A receptor antagonist 8-(3-chlorostyryl)caffeine (CSC) on L-DOPA-induced dopamine (DA) release in the striatum of intact and reserpine-treated rats. CSC given in a pharmacologically effective dose of 5 mg/kg i.p. significantly increased striatal DA release after joint administration of L-DOPA (100 mg/kg, i.p.) and benserazide (50 mg/kg, i.p.) to intact and reserpine (2.5 mg/kg, s.c.)-injected rats. CSC did not change the elevated level of 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) in intact rats, but raised it in DA-depleted animals. The availability of exogenous L-DOPA in the extracellular space was similar and equally increased by CSC in both intact and reserpinized rats. Our results suggest that the observed effects may be mediated by striatal adenosine A2A receptors, and are probably related to the CSC action on DA metabolism and the increased transport of exogenous L-DOPA into the brain. These observations might be of relevance, considering the use of selective A2A antagonists as potential supplements to L-DOPA therapy of Parkinson's disease. PMID- 14751594 TI - Relieving effects of electroacupuncture on mechanical allodynia in neuropathic pain model of inferior caudal trunk injury in rat: mediation by spinal opioid receptors. AB - The relieving effects of electroacupuncture (EA) on mechanical allodynia and its mechanism related to the spinal opioid system were investigated in a rat model of neuropathic pain. To produce neuropathic pain in the tail, the right superior caudal trunk was resected between the S1 and S2 spinal nerves. Two weeks after the surgery, EA stimulation (2 or 100 Hz, 0.3 ms, 0.2-0.3 mA) was delivered to Zusanli (ST36) for 30 min. The degree of mechanical allodynia was evaluated quantitatively by touching the tail with von Frey hair (2.0 g) at 10 min intervals. These rats were then subjected to an i.t. injection with one of the three specific opioid agonists in successive ways: the mu agonist (DAMGO 25, 50 and 100 pmol), the delta agonist (DADELT II 0.5, 1 and 2 nmol), and the kappa agonist (U50488H 5, 10 and 20 nmol) separated by 10 min in cumulative doses. During 30 min of EA stimulation, specific opioid antagonists were subjected to i.t. injection: the mu antagonist (beta-FNA 5, 10 and 20 nmol), the delta antagonist (naltrindole 5, 10 and 20 nmol), and the kappa antagonist (nor-BNI 3, 6 and 12 nmol) separated by 10 min in cumulative doses. As a result, EA reduced the behavioral signs of mechanical allodynia. Two Hz EA induced a robust and longer lasting effect than 100 Hz. All three opioid agonists also showed relieving effects on mechanical allodynia. However, nor-BNI could not block the EA effects on mechanical allodynia, whereas beta-FNA or naltrindole significantly blocked EA effects. These results suggest that the mu and delta, but not kappa, opioid receptors in the spinal cord of the rat, play important roles in mediating relieving effects on mechanical allodynia induced by 2 Hz EA. PMID- 14751595 TI - High-dose aspirin is neuroprotective in a rat focal ischemia model. AB - Acetylsalicylic acid (ASA) is neuroprotective through various pharmacological action sites. We used a temporary middle cerebral artery occlusion (tMCAO) model in 56 Wistar rats to assess whether repeated ASA injections at 30 min, 6 h, 1, 2, 3, and 4 days after stroke onset are neuroprotective. Animals were sacrificed 5 days after MCAO; infarct size was analyzed with 2,3,5-triphenyltetrazolium chloride staining. As compared to saline (164+/-13 mm(3), n=14), only repeated injections of 40 mg/kg ASA (79+/-18 mm(3), n=14, P=0.0029), but not of 20 mg/kg ASA (129+/-19 mm(3), n=15), reduced infarct volume significantly. No significant change was noted with 40 mg/kg ASA injected only once at 30 min after MCAO (117+/ 16 mm(3), n=13). PMID- 14751593 TI - Neutrophils both reduce and increase permeability in a cell culture model of the blood-brain barrier. AB - This study was carried out to determine the effects that human neutrophils have on permeability across a model of the blood-brain barrier (BBB) formed by primary cultures of bovine brain microvessel endothelial cells (BBMEC). Transendothelial electrical resistance (TEER) was used to measure changes in permeability across BBMEC monolayers in a dual compartment system, during neutrophil interactions. When neutrophils (5 x 10(6)/ml) were applied to monolayers, TEER increased (permeability decreased). Adenosine was implicated, since the TEER increase was blocked by adenosine deaminase (1 U/ml) and the adenosine A2 receptor antagonist ZM 241385 (at 10(-6) M but not 10(-8) M, implicating A2B receptors). Oxygen free radicals were implicated as the TEER increase was blocked by combined catalase (100 U/ml) and superoxide dismutase (60 U/ml). When a gradient of the bacterial chemoattractant peptide formyl methionyl leucine phenylalanine (fMLP, 10(-7) M) was applied to neutrophils, the TEER decreased (permeability increased), concurrent with migration. When fMLP (10(-7) M) was added to the neutrophils, without migration, no change occurred. The TEER decrease was blocked by loading endothelium with the calcium buffer BAPTA (10 microM) and partially blocked by the serine protease inhibitor aprotinin (20 microg/ml). Measures to block the potential extracellular triggers heparin binding protein, glutamate, oxygen free radicals and binding to intercellular cell adhesion molecule-1 (ICAM-1) were ineffective. These data indicate that neutrophils both reduce and increase permeability in a cell culture model of the BBB, correlated to their proximity and migration through the endothelium. They explore the role of neutrophils in BBB breakdown, and the formation or amelioration of vasogenic cerebral edema. PMID- 14751596 TI - Delayed post-ischemic albumin treatment neither improves nor worsens the outcome of transient focal cerebral ischemia in rats. AB - Human albumin therapy within the first 4 h is highly neuroprotective in focal ischemia, but it is unknown whether delayed albumin therapy is deleterious. Rats received 2 h middle cerebral artery suture-occlusion. Human albumin (25%, 2.5 mg/kg; n=12) or vehicle (0.9% saline, 5 ml/kg; n=9) were administered at 19 h. Neurological status was evaluated daily, and histopathology and brain swelling were quantified at 3 days. Delayed albumin treatment, while ineffective, failed to show adverse effects. PMID- 14751597 TI - GABA, serotonin and serotonin receptors in the rat inferior colliculus. AB - GABA and serotonin in the mammalian inferior colliculus both have a restrictive effect on fearful and aversive behavior. Immunohistochemical techniques were used to study the relationship between GABA and serotonin receptors in the central nucleus of the rat inferior colliculus. Neurons positive for 5HT1B are more numerous than those displaying 5HT1A receptors. Approximately two-thirds of GABA positive neurons are associated with serotonin receptors. PMID- 14751598 TI - Efficacy of nociceptin inhibition on WDR neuron activity is enhanced in mononeuropathic rats. AB - Nociceptin (NC), administered microiontophoretically at different concentrations, significantly reduced the spontaneous and stimulus-evoked activity on WDR neuron in rats with chronic constriction of one sciatic nerve and showing signs of neuropathic pain. The effect was not antagonized by Naloxone. The same concentrations of NC were ineffective on the noxious stimulus evoked responses of WDR neurons in sham and intact rats. This result indicates a facilitated inhibitory action of NC on nociceptive transmission in this pain model. PMID- 14751603 TI - Appropriate technology for neurosurgery. AB - Though more than 50 years have elapsed since neurosurgery has come to the developing countries, vast areas of the population do not have neurosurgical facilities available to them. This is due not only to the economic status of the country but also as a result of the training that the neurosurgical trainee receives. It is proposed in this article that while the young neurosurgeon must, without doubt, be very well trained in the latest technology, at the same time he must be taught to work with confidence with whatever facilities are available without any feeling of inferiority. The locally available appropriate technology should be fully utilized. The neurosurgeon at the periphery must be provided with the minimum essential neurosurgical equipment and also with good compensation. The concerned governments, the public, neurosurgical teachers, and neurosurgical societies and journals have a great role to play in this difficult task. PMID- 14751611 TI - Quo vadimus: the perspective from neurosurgery's next generation. PMID- 14751612 TI - Evaluation of the Miethke dual- switch valve in patients with normal pressure hydrocephalus. AB - OBJECTIVE: Especially in patients with normal pressure hydrocephalus (NPH), conventional differential-pressure valves are known to create nonphysiological negative intraventricular pressure values (IVP) when the patient moves into the upright position, with the consequence of numerous, sometimes severe, complications. The recently presented gravitational devices promise improvement, primarily in respect to this disadvantage. METHODS: In a prospective multicenter study the new Miethke dual- switch valve (DSV) has been implanted in 128 patients with NPH. The patients have been assessed before operation, at discharge, and re evaluated 6 to 9 months after surgery. The technical principle of the new device is presented. RESULTS: The clinical follow-up showed excellent results in 63% of the patients, satisfactory results in 16%, and a bad outcome in 21% of the cases. The infection rate was 5%; the rate of mechanical complications including overdrainage and dislocations has been in total 9%, and underdrainage was suspected in 7 cases. The outcome correlated with the preoperative severity of NPH. Despite the clinical outcome, the computed tomography scans showed only minimal or no reduction of the ventricular size in the majority of cases. We found a valve-related rate of overdrainage of 2.5%, which is clearly lower than results of comparable series in the literature. CONCLUSION: The clinical course of patients suffering from NPH is mainly influenced by the stage of the disease, the time of beginning of the therapy, and the gravitational function of the implanted device. Based on our clinical experiences with the Miethke dual-switch valve (MD-SV), we underscore the advantages of this valve for the treatment of hydrocephalus, especially for patients with NPH. PMID- 14751619 TI - Sacral tuberculosis: a case report and review of the literature. AB - BACKGROUND: We present a patient with a heterogeneously enhancing lesion within the body of the sacrum and the sacral canal. Sacral tuberculosis (TB) was suspected because of a history of familial exposure. Few cases isolated to the sacrum have been reported in the literature. The characteristic histopathologic and magnetic resonance imaging (MRI) findings are also presented. CASE DESCRIPTION: A 31-year-old African-American male presented with an 8-month history of lower back pain radiating into his legs as well as numbness and weakness of the right foot. His history revealed PPD conversion following an exposure to active pulmonary TB 3 years prior for which he received 6 months of isoniazid prophylaxis. An MRI scan revealed a large heterogeneously enhancing lesion involving the sacrum with extension into the sacral canal. The patient underwent computed tomography (CT)- guided needle biopsy of the sacral lesion. Cytopathologic examination revealed caseating granulomas. Acid-fast bacilli cultures were positive for Mycobacterium tuberculosis. He was placed on a 6-month course of isoniazid, rifampin, pyrazinamide, and ethambutol. At 3-month follow up, his examination and symptoms had improved. CONCLUSIONS: Sacral TB is an extremely rare cause of lower back pain with radiation into the lower extremities. TB should always be considered in the differential diagnosis of isolated sacral masses, especially in light of today's increasing multidrug resistance and immunosuppressed population. PMID- 14751616 TI - Intraoperative antibiotic prophylaxis in clean spinal surgery: a retrospective analysis in a consecutive series of 973 cases. AB - BACKGROUND: Antibiotic prophylaxis in spine surgery is still a debated question, involving medical, ethical, economic, and legal issues. The aim of our retrospective study was to evaluate the safety and effectiveness of an intraoperative protocol of antibiotic prophylaxis. METHODS: During a 3-year period, 973 patients were consecutively operated on for clean spinal operations. Twenty-three percent of the cases involved the cervical spine and 77% the thoraco lumbar spine; about 90% of patients were operated on for degenerative diseases and the remaining for traumatic lesions or tumors. Patients undergoing operations shorter than 120 minutes received a single-dose of IV ampicillin 1000 mg and sulbactam 500 mg (AS) at induction of anesthesia. In procedures longer than 120 minutes and/or requiring prosthetic materials, an IV single-dose of teicoplanin 400 mg was also administered at the same time. A second intraoperative dose of AS and teicoplanin was administered in operations longer than 4 hours (240 minutes after the first one) and in procedures in which blood loss exceeded 1500 mL. Postoperative prophylaxis has never been performed. RESULTS: The only side effect was a cutaneous rash in 7 cases (0.7%), without any consequence. A wound infection was detected in 9 cases (<1%), all successfully treated with surgical toilette and specific antibiotic treatment. A lumbar discitis was detected in 4 out of 657 microdiscectomies (0.6%). CONCLUSIONS: Even if this study has the weakness of the retrospective character, our intraoperative antibiotic prophylaxis protocol proved to be safe and efficacious. We hope that these preliminary results will be confirmed by larger prospectic trials. PMID- 14751624 TI - Atypical presentation of spontaneous discitis: case report. AB - BACKGROUND: Spontaneous discitis is primarily a pediatric illness. Adult patients usually present at an average age of 69 years with a history of diabetes or with a systemic infection. The lumbar spine is the most frequent site of infection (54%), and the cervical is the least at 10%. The causative organisms are most commonly Staphylococcus aureus and beta-hemolytic streptococcus species. Intravenous antibiotics are the mainstays of treatment, and surgical intervention is usually not required. CASE PRESENTATION: A single case observation with an unusual presentation from the statistically typical criterion of discitis is described. CONCLUSIONS: Atypical discitis needs to be considered in the differential diagnoses in the middle-aged and healthy population. PMID- 14751626 TI - Extending survival in gliomas: surgical resection or immunotherapy? PMID- 14751627 TI - The combination of mitotic and Ki-67 indices as a useful method for predicting short-term recurrence of meningiomas. AB - BACKGROUND: The most relevant factor in the progression-free survival (PFS) of patients with meningiomas is the malignant grade. However, using only the current World Health Organization (WHO) definition that does not consider precise quantitative indicators, an unequivocal diagnosis of the malignant grade is difficult. In our retrospective study of the PFS of meningioma patients, we focused on mitoses and the Ki-67 staining index of tumor specimens obtained at the initial surgery. METHODS AND RESULTS: A total of 349 patients with intracranial meningioma, operated between 1978 and 2000, were followed for a mean of 7 years. According to the mitotic index (MI), we classified them into 3 groups. In Group A (n = 326), slide-mounted tumor samples exhibited no mitoses; in Group B (n = 15) there were fewer than 4 mitoses, and in Group C (n = 8) 4 or more mitoses were seen per 10 high-power fields (HPF). The estimated 5-year PFS rates in Groups A, B, and C were 93%, 10%, and 13% respectively. The mean PFS for Group A was 148 months; in Groups B and C the median PFS was 43 and 16 months, respectively. A Ki-67 staining index (SI) of less than 1% corresponded with no mitosis, while an SI exceeding 5% was indicative of the presence of mitoses. CONCLUSION: In meningioma patients, no mitoses and/or a Ki-67 SI <1% signals a favorable outcome. An SI >5% or the presence of mitoses, even fewer than 4 in 10 HPF, is suggestive of a short PFS irrespective of other pathologic features. We suggest that in combination, assay of the Ki-67 SI and the MI represents a reliable, quantitative tool for predicting PFS in meningioma patients. PMID- 14751629 TI - Surgical options in the management of falcotentorial meningiomas: report of 13 cases. AB - BACKGROUND: The primary aim of surgical treatment for falcotentorial meningiomas is gross total excision. The vital surrounding brain structures make this a complex task. Seeking elements that might facilitate presurgical planning we reviewed our surgical records of falcotentorial meningiomas treated during the past 20 years. METHODS: Between 1975 and 1996, in the Neurosurgical Unit at the University of Rome, "La Sapienza," 13 consecutive patients underwent surgery for falcotentorial meningiomas that had been localized on preoperative imaging and confirmed by histology. The surgical approach varied according to the site of the tumor. RESULTS: Nine meningiomas were totally removed and 4 subtotally. Three patients (23.0%), all treated early in the series, died after the operation. Ten patients (76.9%) survived: 3 (23.0%) had postoperative neurologic complications necessitating reintervention, and 7 patients (53.8%) had an uneventful postoperative course. Two of the 4 patients who had subtotal resections had regrowth at 1 year that responded to radiosurgery. CONCLUSIONS: The ideal surgical approach to falcotentorial meningiomas should allow gross total removal and minimum brain retraction while safeguarding the galenic system and other vital neighboring structures. Toward achieving this aim we propose detailed preoperative imaging studies to classify falcotentorial meningiomas according to their site and direction of growth-craniocaudal or anteroposterior-in relation to the cerebellar tentorium. PMID- 14751632 TI - Parasagittal meningioma en plaque with extracranial extension presenting diffuse massive hyperostosis of the skull. AB - BACKGROUND: We describe a case of convexity meningioma en plaque (MEP) invading the skull and scalp with diffuse massive hyperostosis, presenting striking radiological findings. CASE DESCRIPTION: A 48-year-old woman was admitted to our hospital with headache, general fatigue, diplopia, and blurred vision. A skull X ray film, computed tomography (CT), and 3-dimensional CT demonstrated diffuse significant hyperostosis and sclerosis in the cranial vault. Magnetic resonance (MR) imaging revealed a frontoparietal, well-enhanced extra-axial mass lesion, which is compatible with findings of MEP. The tumor was partially resected. Histological examination revealed massive tumor invasion into the dura mater, hyperostotic skull, and scalp. CONCLUSION: Hyperostosis is frequently observed in MEP, although there has been no report of a case of MEP such as ours demonstrating diffuse tumor extension and diffuse hyperostosis. Analyzing the CT finding of hyperostotic bone is useful for differentiation of MEP from other diseases that include a hyperostotic condition. PMID- 14751635 TI - Unmasking the secrets of meningioma: a slow but rewarding journey. PMID- 14751636 TI - Solitary cerebellar metastasis of malignant pleural mesothelioma: case report. AB - BACKGROUND: Malignant pleural mesothelioma is an uncommon malignancy that rarely metastasizes to the central nervous system and even less frequently occurs as a solitary lesion. CASE DESCRIPTION: We present a 71-year-old white female, nonsmoker, with no occupational exposure to asbestos. She presented with a 15-lb. weight loss over several months and persistent right subscapular pain radiating to her anterior chest. Imaging studies revealed a pleural mass, and biopsy confirmed fibrous type malignant pleural mesothelioma. During a metastatic workup, computed tomography (CT) and magnetic resonance imaging (MRI) of the head demonstrated a 1 cm subcortical, contrast-enhancing lesion without surrounding edema in the right posterior cerebellum. Surgical resection of the solitary cerebellar mass revealed fibrous-type metastatic malignant mesothelioma. Postoperatively, the patient received a combined chemotherapy regimen of Adriamycin and Cisplatin and underwent whole brain radiation therapy. CONCLUSIONS: We report the first resection of a solitary cerebellar metastasis of malignant pleural mesothelioma. We also review past cases of intracranial metastasis of this malignancy, its histologic subtypes, outcome, and recent treatment modalities. PMID- 14751638 TI - Development of glioblastoma multiforme following traumatic cerebral contusion: case report and review of literature. AB - BACKGROUND: Diagnostic criteria for posttraumatic brain tumors were formulated in the pre-CT (computerized tomography) era. We propose that radiologic criteria incorporating imaging data be added to the existing criteria. CASE DESCRIPTION: We report a case of a 56-year-old man who presented with history of raised intracranial pressure of 20 days' duration. Imaging showed a large left frontal intra-axial mass lesion. He had history of head injury 5 years prior with CT evidence of bilateral basifrontal contusions. There was no contrast enhancement at the site of the contusions in an intervening CT scan done 18 months after the trauma. He underwent radical excision of the mass, and the histopathology was reported as glioblastoma multiforme. We formulated additional radiologic criteria for tumors that may present following trauma. A review of the literature of posttraumatic gliomas is also presented. CONCLUSION: Fulfillment of the additional radiologic criteria proposed by us will help distinguish a tumor that developed following trauma from that which was present before the occurrence of the injury. PMID- 14751640 TI - Treatment of vertebral artery aneurysms with posterior inferior cerebellar artery posterior inferior cerebellar artery anastomosis combined with parent artery occlusion. AB - BACKGROUND: In patients with aneurysms that involve the origin of the posterior inferior cerebellar artery (PICA) and require occlusion of the vertebral artery (VA), revascularization of the PICA is commonly performed. We present six patients with dissecting VA aneurysms who underwent PICA-PICA anastomosis combined with parent artery occlusion. METHODS: After a lower lateral suboccipital craniectomy and partial resection of the jugular tubercle, anastomoses were performed in a side-to-side fashion at the posterior medullary segment of the PICA. The VA was subsequently occluded by clipping proximal and distal to the aneurysm, and the PICA was occluded by clipping distal to the aneurysm. RESULTS: Postoperative cerebral angiography demonstrated patency of the anastomosis and regression of the aneurysm in five of six patients. The remaining patient experienced hemorrhage from contralateral VA dissection and subsequently died. One patient experienced myopathy of the lower extremities secondary to intraoperative fixed board compression and developed permanent lower extremity muscular weakness. The remaining four cases experienced no new neurologic deficits. CONCLUSION: PICA-PICA anastomosis is a useful procedure for reconstruction of the PICA when parent vessel occlusion or trapping is necessary to exclude a VA aneurysm involving the origin of the PICA. PMID- 14751644 TI - Venous angioma of the facial nerve: a case report. AB - BACKGROUND: Lower motor neurone type facial nerve palsy is a common condition that is very rarely caused by a neoplastic lesion. CASE DESCRIPTION: This case report describes a progressive facial palsy in a 62-year-old man because of a vascular tumor of the facial nerve. Histologic examination confirmed it to be a venous angioma. CONCLUSION: Meticulous imaging of the facial nerve is required to diagnose cases of progressive facial nerve palsy caused by a neoplastic lesion. PMID- 14751642 TI - Microsurgical anatomy of the perforating branches of the vertebral artery. AB - BACKGROUND: There is limited data in the literature related to the microanatomic features of the perforating branches of the vertebral artery. METHODS: The 44 vertebral arteries and their branches were injected with india ink or a radiopaque substance and examined under the stereoscopic microscope. RESULTS: The perforating arteries were noted to range in number from 1 to 11 (mean, 6.5) and in diameter between 100 microm and 520 microm (average, 243 microm). They arose from the vertebral artery (VA) (54.54%), 8 from the right, the left or both VAs. The anterior spinal artery (ASA), which was singular (81.82%), duplicated (13.64%), or plexiform (4.55%), always gave rise to the perforators. The vascular roots of the ASA were the source of the perforators in 95.45% of the brains. The latter vessels arose from the anterolateral arteries in 50% of the cases. The anastomoses involving the perforators, which were present in 40.91% of the brains, varied in diameter between 100 microm and 350 microm (mean, 169 microm). The perforating vessels gave rise to the side branches in 95.45% of the brains that varied in diameter from 100 microm to 300 microm (average, 161 microm). The perforators usually entered the foramen cecum and the anterior median sulcus, and then continued close and parallel to the raphe of the medulla. The perforators can be compressed by a VA aneurysm, which was found in one among the 71 examined patients with cerebral aneurysms. CONCLUSIONS: The obtained data give additional information about the vascular anatomy of the pontomedullary region. PMID- 14751645 TI - Split spinal cord malformation in an elderly patient: case report. AB - Split spinal cord malformations (SCM) typically present in childhood and rarely in adulthood. Very little is known about the SCMs in the elderly, and the diagnosis can be easily missed. A 73-year-old woman with a childhood history of scoliosis and late ambulation milestones presented with a 2-year history of worsening low back pain and progressive difficulty walking. She had a mild gait disturbance with 4/5 weakness in left ankle dorsiflexion. Magnetic resonance imaging revealed a bifid spinal cord contained in a single thecal sac and a tethered cord with low-lying conus at L3. The patient was taken to the operating room and a soft-tissue median septum, as well as all other adhesions, was removed. The filum terminale was identified, coagulated, and divided. Six weeks later, the patient reported decreased back pain, improvement in ambulation, and markedly decreased used of narcotics for her back and leg pain. Her left ankle dorsiflexion strength improved to 4+/5. This patient had two hemicords encased in a single dural tube separated by a nonrigid, fibrous median septum and an associated tethered cord. Adult presentation of SCM is extremely rare. This case highlights the need to consider split cord malformation and tethered cord in the differential diagnosis not only for adults but also the elderly presenting with back pain, scoliosis, and difficulty walking. PMID- 14751647 TI - The National Institute of Neurology and Neurosurgery of Mexico on its 40th anniversary. PMID- 14751648 TI - Where has the concern for humanity gone? PMID- 14751650 TI - Omapatrilat and enalapril in patients with hypertension: the Omapatrilat Cardiovascular Treatment vs. Enalapril (OCTAVE) trial. AB - BACKGROUND: Recent reports suggest that existing antihypertensive agents may not have sufficient efficacy to control blood pressure (BP) in many patients. Omapatrilat, an agent under development, has been shown to have significantly greater antihypertensive efficacy than existing agents, but may also carry increased risk of angioedema. We compared the efficacy and safety of omapatrilat to a representative angiotensin-converting enzyme (ACE) inhibitor, enalapril. METHODS: The Omapatrilat Cardiovascular Treatment vs. Enalapril (OCTAVE) trial is a multicenter, randomized, double-blind, active-controlled, 24-week trial in 25,302 patients with untreated or uncontrolled hypertension conducted in 3298 office-based sites in 12 countries. Subjects were randomized to omapatrilat 10 mg or enalapril 5 mg as initial therapy for hypertension (group 1, n = 9292), replacement for existing antihypertensive therapy (group 2, n =11,224), or in addition to existing antihypertensive therapy (group 3, n = 4751). Study drug was force-titrated at week 2 and electively titrated at weeks 4 and 6 to a maximum of omapatrilat 80 mg or enalapril 40 mg once daily. At weeks 8 and 16, adjunctive antihypertensive medications were added electively to achieve target BP. RESULTS: Omapatrilat reduced systolic BP 3.6 mm Hg more than enalapril at week 8, and was associated with less use of adjunctive antihypertensive therapy by week 24 (19% v 27%; P < 0.001 for both comparisons). Subjects randomized to omapatrilat were more likely to reach BP target, regardless of demographics or comorbid conditions and whether omapatrilat was used as initial therapy, replacement for existing therapy, or in addition to existing therapy. Angioedema was more frequent with omapatrilat than enalapril (2.17% v 0.68%). Two omapatrilat-treated subjects experienced angioedema with airway compromise, which was successfully treated. CONCLUSIONS: Omapatrilat provided broadly superior antihypertensive efficacy when used in a setting resembling clinical practice. Angioedema was more common than with enalapril but life-threatening angioedema was rare. The risk-benefit profile for omapatrilat in clinical use therefore appears likely to be favorable in appropriate patients. PMID- 14751651 TI - Comparison of valsartan and amlodipine on ambulatory and morning blood pressure in hypertensive patients. AB - BACKGROUND: Cardiovascular events occur most frequently in the morning. We aimed to study the effects of monotherapy with the long-acting angiotensin II receptor blocker valsartan compared with the long-acting calcium antagonist amlodipine on ambulatory and morning blood pressure (BP). METHODS: We performed ambulatory BP monitoring before and after once-daily dose of valsartan (valsartan group, n = 38) and amlodipine (amlodipine group, n = 38) therapy in 76 hypertensive patients. To achieve the target BP of < or =140/90 mm Hg, valsartan was titrated from 40 mg/day to 160 mg/day (mean dose 124 mg/day) and amlodipine was titrated from 2.5 mg/day to 10 mg/day (mean dose 6.4 mg/day). RESULTS: Both drugs significantly reduced clinic and 24-h systolic BP (SBP) and diastolic BP (DBP) (P <.002). However, the antihypertensive effect of amlodipine was superior to that of valsartan in clinical SBP (-26 mm Hg v -13 mm Hg, P =.001) and 24-h SBP (-14 mm Hg v -7 mm Hg, P =.008). In addition, morning SBP was significantly reduced by amlodipine from 156 to 142 mm Hg (P <.001) but not by valsartan. Both agents reduced lowest night SBP to a similar extent (amlodipine 121 to 112 mm Hg, P <.001; valsartan 123 to 114 mm Hg, P <.002). Reduction in morning SBP surge (morning SBP minus lowest night SBP) was significantly greater in patients treated with amlodipine compared with those treated with valsartan (-6.1 mm Hg v +4.5 mm Hg, P <.02). CONCLUSIONS: Amlodipine monotherapy was more effective than valsartan monotherapy in controlling 24-h ambulatory BP and morning BP in hypertensive patients. PMID- 14751652 TI - Effect of different antihypertensive drug classes on central aortic pressure. AB - Central aortic systolic blood pressure (BP) is an important determinant of cardiac workload and cardiac hypertrophy. The relationship of central aortic systolic BP and brachial BP varies depending on the stiffness of blood vessels. It is not certain whether the different drug classes affect the brachial and aortic systolic BP in a similar manner. In a double-blind crossover study, we measured the effects of the four major drug classes compared with placebo on central aortic pressure. Central aortic pressure and various indices were determined using the Sphygmo Cor apparatus. The study was undertaken in patients aged 65 to 85 years with systolic BP >150 mm Hg at study entry. Results are reported for 32 patients who had satisfactory applanation tonometry in all five periods. Calcium channel blockers and diuretics caused a greater fall in brachial artery systolic BP than angiotensin-converting enzyme (ACE) inhibitors or beta blocking drugs. On placebo, central aorta augmentation pressure and index were 23 mm Hg and 33.3%; on ACE inhibitors the values were 18 mm Hg and 30%; on beta blockers, 26 mm Hg and 38.5%; on calcium channel blockers, 16 mm Hg and 28%; and on diuretics, 17 mm Hg and 28.8%. The augmentation pressure on beta-blocking drugs was greater than on the other three drug classes (P <.05), and augmentation pressures on ACE inhibitors, calcium channel blockers, and diuretics were less than on placebo (P <.05). The lowest central aortic pressures were achieved with calcium blocking drugs and diuretics. Therapy based on brachial artery recordings may thus overestimate the effect of beta-blocking drugs on central aortic systolic BP and underestimate the effectiveness of ACE inhibitors and calcium blocking drugs. The clinical importance of this discrepancy needs to be evaluated. PMID- 14751653 TI - White coat effect in treated versus untreated hypertensive individuals: a case control study using ambulatory and home blood pressure monitoring. AB - BACKGROUND: Some studies have shown a significant white coat effect (WCE) (i.e., difference between clinic blood pressure [CBP] and awake ambulatory blood pressure [ABP]) to be present not only in untreated but also in treated hypertensive individuals. This study aims to assess 1) the prevalence and the magnitude of the WCE in treated versus untreated hypertensive persons, and 2) the usefulness of home blood pressure (HBP) versus ABP in the detection of this phenomenon. METHODS: A case-control study was conducted in 138 treated hypertensive patients and same number of sex- and age-matched untreated hypertensive subjects who had measurements of CBP (at least three visits), HBP, and ABP. Subjects with a WCE of >20/10 mm Hg (systolic/diastolic) were classified as clinic reactors. RESULTS: There was a trend for a larger WCE assessed by ABP monitoring in the untreated group (mean difference in systolic WCE, 1.8 +/- 22.2 mm Hg, 95% CI -2.0 to 5.5; diastolic 1.8 +/- 11.9 mm Hg, 95% CI -0.2 to 3.8) and for more untreated clinic reactors (27% untreated v 20% treated, odds ratio 1.5, 95% CI 0.9 to 2.7). The sensitivity, specificity, and positive and negative predictive values of HBP to detect clinic reactors correctly were 56%/62% (treated/untreated), 87%/84%, 52%/59%, and 89%/86%, respectively, with moderate agreement between HBP and ABP (kappa 0.42/0.46). CONCLUSIONS: In treated hypertensive patients, WCE seems to be reduced compared with that in untreated hypertensive persons but is not eliminated. In both untreated and treated hypertensive individuals HBP monitoring appears to be useful in the detection of the WCE, but it may not be appropriate as an alternative to the ABP method. PMID- 14751654 TI - Aortic pulse pressure is related to the presence and extent of coronary artery disease in men undergoing diagnostic coronary angiography: a multicenter study. AB - BACKGROUND: In the general population, pulse pressure (PP) is a correlate of cardiovascular outcomes. Few data are available regarding the links between PP and documented coronary artery disease (CAD). METHODS: From July 2000 to January 2002, a total of 1337 patients referred for a first diagnostic coronary angiogram at 75 participating centers were prospectively included. Of these individuals, 280 patients receiving no hypertensive therapy constituted the study population. Pulse pressure was recorded in the aortic root before angiography, and baseline characteristics, medical history, treatment used, and data from coronary angiography were recorded. RESULTS: In the whole population, aortic PP strongly correlated with the presence and extent of CAD in univariate analyses. However, the correlation disappeared in multivariate analysis, and a strong interaction with gender was found. In women (n = 82), aortic PP was not an independent predictor of CAD. However, in men (n = 198) an independent correlation between aortic PP and CAD was found, together with age and hypercholesterolemia. In addition, PP was strongly correlated with the extent of CAD (no disease, 51 +/- 16 mm Hg; one or two stenoses, 54 +/- 18 mm Hg; and more than two stenoses: 64 +/ 20 mm Hg). CONCLUSIONS: In this multicenter study, aortic PP was significantly correlated with the presence and extent of CAD in patients without antihypertensive therapy. This correlation, however, was independent of other risk factors for CAD in men but not in women. PMID- 14751655 TI - Clinical experience with perindopril in African-American hypertensive patients: a large United States community trial. AB - BACKGROUND: The prevalence of hypertension is greater in African Americans, and management of this condition presents challenges for practicing physicians. METHODS: The effectiveness and safety of perindopril was evaluated in hypertensive African-American patients (n = 1412) and hypertensive white patients (n = 7745) who had participated in a large United States community trial. Patients received perindopril 4 mg once daily for 6 weeks. Based on physicians' clinical judgment at week 6, the dose was either maintained or increased to 8 mg for an additional 6 weeks. RESULTS: Reduction of blood pressure (BP) was significant with perindopril monotherapy (4 to 8 mg once daily) in African Americans and whites (P <.001). The magnitude of BP reduction was significantly more in whites (P <.001). Up-titration of perindopril achieved additional BP reduction in both ethnic groups (P <.001). Control of BP (<140/90 mm Hg) in elderly (>65 years of age) and diabetic African-Americans subgroups was achieved in 32.1% and 31.6%, respectively. Perindopril was safe and well tolerated. CONCLUSIONS: Perindopril monotherapy is effective and is a viable initial therapeutic option as an antihypertensive agent in African-American individuals with hypertension. PMID- 14751656 TI - Erectile dysfunction in essential arterial hypertension and effects of sildenafil: results of a Spanish national study. AB - BACKGROUND: The aim of this study was to establish the prevalence of erectile dysfunction (ED) in hypertensive patients in specialized care hypertension units (SCHUs) and to assess the effectiveness and tolerability of sildenafil treatment. METHODS: This was a multicenter, prospective, open, observational pharmacoepidemiology study conducted in 25 Spanish SCHUs. A total of 2130 men with essential hypertension under treatment were recruited. In a second phase, 291 subjects with a score < or = 21 in the Sexual Health Inventory for Men (SHIM) received sildenafil (50 mg/day) as required 30 to 60 minutes before sexual activity, and were evaluated by the International Index of Erectile Function (IIEF). RESULTS: A total of 975 subjects (45.8%) had a score < or = 21 in the SHIM. In the second phase, sildenafil improved the score in the erectile function domain in 232 patients (83.2%). Severity of ED significantly improved (P <.001); severe (22.3% to 7.7%), moderate (23% to 5.6%), and mild impairment (36.3% to 44.8%). The IIEF was normalized in 39.1% of patients who completed post-treatment IIEF. In all, 33 subjects (11.8%) failed to complete the study: two (0.7%) because of lack of efficacy, two (0.7%) intercurrent disease, 10 (3.6%) failure to return to the visits, three (1.1%) fear of therapy, four (1.4%) adverse effects requiring treatment discontinuation, and 12 (4.3%) protocol violations. No statistically significant association was found between the prevalence of adverse effects and antihypertensive treatment with single drug or combination therapy. CONCLUSIONS: A high incidence of ED was found in hypertensive patients from Spanish SCHUs. Sildenafil showed an excellent response and safety profile. PMID- 14751657 TI - Glucose effectiveness is strongly related to left ventricular mass in subjects with stage I hypertension or high-normal blood pressure. AB - BACKGROUND: Evidence suggests that "glucose effectiveness," (SG) or the effect of glucose per se to enhance net glucose disposal, may be at least as important as the insulin sensitivity index (SI) in the assessment of glucose tolerance. Our objective was to study the relationship of SG and SI parameters to left ventricular mass in a group of untreated, nondiabetic, and nonobese subjects recently diagnosed with stage I or high-normal blood pressure (BP). METHODS: In this sample of subjects, among whom the expected prevalence of insulin resistance is low, we assessed SG and SI parameters using the intravenous glucose tolerance test and minimal model analysis. We also measured left ventricular mass (LVM) index and diastolic function by echocardiography. RESULTS: We observed a strong relationship between SG and LVM index (r = -0.61, P <.0001). Patients with left ventricular hypertrophy (LVH) had lower SG than those without LVH (0.1114 +/- 0.04 v 0.2088 +/- 0.08 x 10(-1). min(-1), P <.001). In contrast, patients below the lowest quartile of the SG parameter distribution had higher LVM index (126.4 +/- 23.1 v 94.8 +/- 22.3 g/m(2), P <.001) and also had higher prevalence of LVH than the other patients (P <.0001). The SI related only to diastolic dysfunction, suggesting that SG may be an earlier marker of LVH than SI in hypertension. CONCLUSION: In this sample of nonobese and glucose-tolerant subjects with an early stage of hypertension, SG but not SI was related to LVM. PMID- 14751658 TI - A prospective study of maternal serum C-reactive protein concentrations and risk of preeclampsia. AB - BACKGROUND: We measured C-reactive protein (CRP), a clinical marker of systemic inflammation, in maternal serum collected at 13 weeks gestation on average, to determine whether elevations precede the clinical manifestation of preeclampsia. METHODS: Using a prospective, nested, case-control study design we measured CRP concentrations using a competitive immunoassay in 60 women who developed preeclampsia and in 506 women who remained normotensive throughout pregnancy. Logistic regression procedures were used to calculate odds ratio (OR) and 95% CI. Because maternal serum CRP is highly correlated with maternal prepregnancy body mass index (BMI), all analyses were repeated after stratification by maternal prepregnancy overweight status (BMI <25 v > or =25 kg/m(2)). RESULTS: Overall, the risk of preeclampsia increased across successively higher tertiles of CRP (OR = 1.0, 1.6, and 3.5, with the lowest tertile as the referent group; P <.001 for trend). After adjusting for parity and first-degree family history of chronic hypertension, the OR in the highest tertile was 3.2 (95% CI = 1.5 to 6.7). Further adjustment for BMI greatly attenuated this association (OR = 1.8, 95% CI = 0.8 to 4.1). Elevated CRP concentrations (> or =4.9 mg/L) were associated with a 2.5-fold increased risk of preeclampsia (95% CI = 1.1 to 5.5) in lean women. No similar association was observed among overweight women. CONCLUSIONS: Elevated CRP is highly correlated with prepregnancy adiposity and appears to be an independent predictor of preeclampsia in lean women. Further work is needed to identify modifiable risk factors for systemic inflammation in early pregnancy and to explore further the extent to which CRP and prepregnancy adiposity independently and jointly contribute to preeclampsia risk. PMID- 14751659 TI - Effects of early carvedilol treatment and withdrawal on the development of hypertension and renal vascular narrowing. AB - BACKGROUND: The aims of this study were to examine whether combined blockade of alpha(1) and beta-adrenoceptors with carvedilol postweaning affected the development of hypertension and renal vascular narrowing in spontaneously hypertensive rats (SHR), and whether these effects on pressure and renal vascular changes persisted after treatment withdrawal. METHODS: From 4 to 12 weeks of age male SHR were administered carvedilol in rat chow at 1.2 mg/g chow (low-dose) or 2.4 mg/g chow (high-dose), or were given normal chow. At 12 weeks of age, rats from each group either underwent experimentation or had treatment withdrawn and were studied at 20 weeks. On the experimental day, conscious mean arterial pressure (MAP) was measured and, as a functional test of renal vessel lumen characteristics, pressure-flow and pressure-glomerular filtration rate (pressure GFR) relationships were determined in the maximally dilated kidney. RESULTS: At 12 weeks of age, SHR on low and high-dose carvedilol had significantly lower MAP than that of untreated SHR (137 +/- 3, 134 +/- 1, 152 +/- 2 mm Hg, respectively; P <.001). The SHR treated with high-dose (but not low-dose) carvedilol demonstrated a steeper renal pressure-flow relationship (P <.001), and a leftward shifted (P <.01) and steeper (P <.001) pressure-GFR relationship compared with control SHR. Eight weeks after carvedilol withdrawal, there were no significant differences in MAP, pressure-flow, or pressure-GFR relationships between groups. CONCLUSIONS: These results suggest that postweaning alpha(1) and beta adrenoceptor blockade with high-dose carvedilol attenuated the development of hypertension and led to a preferential reduction in preglomerular resistance (increased lumen dimensions) independent of the effects on MAP. However, treatment of SHR from 4 to 12 weeks of age with high-dose carvedilol did not lead to persistent, long-term effects on arterial pressure or renal vascular narrowing after treatment withdrawal. PMID- 14751660 TI - Inhibition of oxidative stress and improvement of endothelial function by amlodipine in angiotensin II-infused rats. AB - BACKGROUND: Calcium channel blockers such as amlodipine are effective antihypertensive agents. In this study we investigated the effects of amlodipine on vascular oxidative stress, expression of the lectin-like oxidized low-density lipoprotein receptor (LOX-1), and endothelial function in angiotensin (Ang) II infused rats. METHODS: Sprague-Dawley rats were treated with Ang II (0.7 mg/kg/day subcutaneously injected by mini-pump), with or without amlodipine (10 mg/kg/day by gavage), for 5 days and compared with control rats. Levels of aortic ring superoxide (O(2)(-)) and peroxynitrite (ONOO(-)) were determined, and systolic blood pressure (SBP) and endothelium-dependent relaxation were evaluated. RESULTS: Compared with control rats, Ang II-infused rats developed hypertension (175 +/- 3 v 135 +/- 2 mm Hg, P <.05), aortic hypertrophy (16.9 +/- 1.3 v 13.2 +/- 0.3 mg/cm, P <.05), left ventricular hypertrophy (0.236 +/- 0.003 v 0.204 +/- 0.004 g/100 g body weight, P <.05), and impaired endothelium dependent relaxation (ED(50): 6.6 +/- 0.2 v 8.0 +/- 0.2 -log mol/L acetylcholine concentration, P <.05). Compared with control rats, Ang II-infused rats also had higher aortic levels of LOX-1 mRNA expression, O(2)(-)production (1005 +/- 140 v 608 +/- 159 counts/min/mg, P <.05), ONOO(-) production (1875 +/- 295 v 782 +/- 115 counts/min/mg, P <.05), and plasma free 8-F(2)alpha-isoprostanes (67.4 +/- 19.1 v 27.2 +/- 6.1 pg/mL, P <.05). In Ang II-infused rats SBP, aortic hypertrophy, endothelial dysfunction, LOX-1 expression, aortic O(2)(-) and ONOO( ) production, and plasma free 8-F(2)alpha-isoprostane levels were significantly reduced by amlodipine treatment. CONCLUSIONS: Amlodipine has antihypertensive and antioxidant activity in vivo, which effectively inhibits many of the oxidative stress-dependent mechanisms involved in Ang II-mediated cardiovascular injury. PMID- 14751661 TI - Angiotensin-converting enzyme inhibition and angiogenesis in myocardium of obese Zucker rats. AB - BACKGROUND: Obesity, hypertension, and non-insulin-dependent diabetes mellitus (NIDDM) are associated with microvascular rarefaction in the myocardium and this contributes to increase cardiovascular morbidity and mortality. At present, controversial data exist in medical literature regarding the specific role of angiotensin-converting enzyme (ACE) inhibitors concerning angiogenesis in different tissues. The present study was designed to determine the possible beneficial effects of an ACE inhibitor perindopril on myocardial angiogenesis in an animal model of obesity, hypertension, and NIDDM, such as the obese Zucker rat (OZR) and control lean Zucker rats (LZR). METHODS AND RESULTS: Ten-week-old male OZR (fa/fa) and LZR (Fa/fa) were used in this study: OZR group (G1, n = 10), OZR with perindopril group (G2, n = 10); LZR group (G3, n = 10). For 6 months, G2 received a daily dose of 3 mg/kg of perindopril, by gavage, and G1 and G3 received an equal volume of vehicle throughout the experiment. After 6 months of treatment, all rats were killed, hearts were harvested for pathology studies, including immunohistochemistry, using monoclonal antibodies against rat endothelial cell (RECA-1) and endothelial nitric oxide synthase. At the end of the study, OZR treated with perindopril presented: 1) lower blood pressure (BP) (127 +/- 3.2 v 152.4 +/- 3 mm Hg, P <.01) and 2) lower heart weight/100 g body weight (0.22 +/- 0.02 v 0.36 +/- 0.04 g, P <.01) than OZR untreated. Moreover, OZR that received perindopril showed higher: 1) myocyte density (2044 +/- 67 v 847 +/- 91 myocytes/mm(2), P <.01) and 2) capillary density (1348 +/- 118 v 436 +/- 78 capillaries/mm(2), P <.01); higher amount of: 1) vascular endothelial growth factor (VEGF) in the myocardium (P <.01) and higher percentage of capillaries with positive immunostaining for eNOS (P <.01), compared with untreated OZR. There was a correlation between both the amount of VEGF in myocardium and the number of capillaries (r = 0.88; P <.01) and VEGF and eNOS expression in myocardial capillaries (r = 0.93; P <.01) in OZR treated with perindopril. Finally, OZR that received P showed an improvement in insulin/glucose ratio (P <.01) when compared with untreated OZR. CONCLUSIONS: These results suggest that ACE inhibition by perindopril improves myocardial angiogenesis in this animal model of human metabolic syndrome. The pathway that involves bradykinin, eNOS, and VEGF could be involved in this effect; however, because no additional antihypertensive treatment group was included in our study, the BP-lowering effect cannot be excluded. PMID- 14751662 TI - Characteristic change in local pulse wave velocity in different segments of the atherosclerotic aorta in KHC rabbits. AB - BACKGROUND: Pulse wave velocity, conventionally determined between the carotid and femoral arteries, is a useful measure to estimate stiffness of the aorta. We investigated local pulse wave velocity (LPWV) in different segments in the aorta with relatively early-stage atherosclerosis in relation to the extent and severity of atherosclerotic lesions. METHODS: Pressure waves were recorded in eight aortic positions using two catheters with one or two micromanometers to determine LPWV in the ascending aorta, distal end of the aortic arch, proximal, middle, and distal thoracic aortas, and proximal, middle, and distal abdominal aortas in Kurosawa and Kusanagi-hypercholesterolemic (KHC) and normal rabbits aged 10 to 12 months. RESULTS: The LPWV in the KHC rabbit was greatest in the aortic arch, decreased almost to the normal level in the middle and distal thoracic aorta, increased in the proximal abdominal aorta, and showed almost identical change to that in the normal rabbit in the middle and distal abdominal aortic regions. There was significant difference in LPWV in the aortic arch, proximal thoracic, and proximal abdominal aortas between the two rabbit groups. The sclerotic lesion was prominent in the aortic arch, proximal thoracic aorta, and proximal abdominal aortas. The wall was severely thickened with abundant foam cells. The significant increase in LPWV would be mainly related to the increased wall thickness in these aortic regions. CONCLUSIONS: We can conclude that LPWV reflects well the distribution and severity of atherosclerotic lesion and the increased wall thickness in the local aortic region in which pulse waves were traveled. PMID- 14751663 TI - Mice lacking osteopontin exhibit increased left ventricular dilation and reduced fibrosis after aldosterone infusion. AB - BACKGROUND: Osteopontin, also known as cytokine Eta-1, plays an important role in postmyocardial infarction remodeling by regulating collagen accumulation. Aldosterone promotes collagen synthesis and structural remodeling of the heart. The role of osteopontin in aldosterone-induced fibrosis and myocardial remodeling is unknown. Osteopontin expression and left ventricular structural and functional remodeling were determined in wild-type and osteopontin knockout mice after aldosterone infusion. METHODS AND RESULTS: Immunohistochemical analyses showed increased interstitial osteopontin protein in the wild-type left ventricle after 7 days of aldosterone infusion. After 4 weeks of aldosterone infusion, heart rate was unchanged, and there were similar increases in blood pressure (BP) and heart to-body weight ratio in both wild-type and knockout mice. Left ventricular end diastolic diameter was significantly higher, whereas percent fractional shortening was significantly lower (P <.05) in knockout versus wild-type mice after 4 weeks of aldosterone infusion. Aldosterone infusion increased fibrosis and apoptosis (TUNEL-positive) in both wild-type and knockout mice. However, the increase in the extent of fibrosis and apoptosis was significantly lower in knockout hearts. CONCLUSIONS: Increased osteopontin plays an important role in the regulation of aldosterone-induced remodeling with effects on left ventricular dilation, fibrosis, and apoptosis. PMID- 14751664 TI - Time interval between pairs of arterial blood pressure measurements--does it matter? AB - Guidelines on arterial blood pressure (BP) measurements recommend a time interval between readings. There has been little evidence supporting this recommendation. Therefore, we measured the variation in arterial pressure between the pairs of measurements without an interval (n = 269) or with venous congestion interposed (n = 79). The variation in BP readings was not significantly greater whether a 60 sec interval or venous congestion were present. These overall results do not support the hypothesis that the absence of the time interval or of presence of venous congestion, as may occur in clinical practice, significantly affects BP measurements. PMID- 14751665 TI - In the search for stroke genes: a long and winding road. AB - In spite of a significant improvement in control of numerous predisposing risk factors, stroke remains a major health problem and a common cause of death and disability in our societies. Genetic predisposition to stroke development exists and has been documented in both animal models and in humans. However, a precise definition of genetic factors responsible for common forms of stroke is still lacking, mainly due to its complex nature, the confounding presence of other predisposing risk factors, and the genetic heterogeneity of human populations. In contrast, important breakthroughs have been reached for monogenic forms of stroke, such as cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL). An animal model of stroke, the stroke prone spontaneously hypertensive rat, has provided valuable information on genetic factors involved in stroke predisposition. Among them, the gene-encoding atrial natriuretic peptide has been identified as a stroke gene in both the stroke-prone spontaneously hypertensive rat and, subsequently, in two different human populations. In particular, structural alterations of the gene are consistently present in diseased individuals, suggesting an important role of mutation-dependent mechanisms in stroke predisposition. Finally, the recent use of intermediate disease phenotypes provides a reductionist approach that may contribute to important accumulating information on genes contributing to cerebrovascular accidents. PMID- 14751666 TI - The selective effect of genistein on the toxicity of bleomycin in normal lymphocytes and HL-60 cells. AB - This study was carried out to find whether genistein might enhance bleomycin induced cytotoxicity in human leukemia (HL-60) while protecting normal blood lymphocytes. Despite the excellent chemotherapeutic effect of bleomycin, its cytotoxicity and genotoxicity in normal cells remains a major problem in chemotherapy. Genistein, one of the major Soy isoflavones, is particularly effective in quenching free radicals generated by toxic agents. In this study, the protective and enhancement effects of genistein on bleomycin induced cytotoxicity in HL-60 cells and blood lymphocytes were demonstrated. HL-60 cells were treated with various concentrations of genistein for 3 h followed by treatment with various concentrations of bleomycin during the G1 phase. Pretreatment of genistein increased micronuclei (MN) frequency and DNA damage as a result of bleomycin treatment. However, when human lymphocytes were pretreated with genistein prior to bleomycin treatment during the G2 or G0 phase, the frequencies of bleomycin induced MN was decreased. Although the extent of bleomycin induced DNA damage determined by single cell gel electrophoresis was increased through the pretreatment of genistein in HL-60 cells, it was decreased in normal lymphocytes. The result of this study may therefore provide great impact on the potential activity of genistein as a therapeutic agent. PMID- 14751667 TI - Assessment of oxidative stress in lungs from sheep after inhalation of wood smoke. AB - To elucidate potential dose-dependent mechanisms associated with wood smoke inhalation injury, the present study evaluated antioxidant status and the extent of pulmonary injury in sheep after graded exposure to smoke. Adult, male sheep (n=4-5 per group) were anesthetized and received 0, 5, 10 or 16 units of cooled western pine bark smoke, corresponding to 0, 175, 350 and 560 s, respectively, of smoke dwell time in the airways and lung. Smoke was mixed at a 1:1 ratio with 100% O2 to minimize hypoxia. Plasma and expired breath samples were collected pre smoke, and 6, 12, 18, 24, 36 and 48 h after smoke exposure. Sheep were euthanatized 48 h after smoke exposure and lung and airway sections were evaluated histologically for injury and biochemically for indices of oxidative stress. Plasma thiobarbituric acid reactive substances (TBARS) were 66 and 69% higher than controls after moderate and severe smoke exposure at 48 h, whereas total antioxidant potential was not statistically different among groups at any time after exposure. Lung TBARS showed a dose-dependent response to smoke inhalation and were approximately 2-, 3- and 4-fold higher, respectively, than controls after exposure to 5, 10 and 16 units of smoke. Lung myeloperoxidase (MPO) activity was also higher in smoke-exposed animals than controls, and MPO activity was markedly elevated (19- and 22-fold higher than controls in right apical and medial lobes) in response to severe smoke exposure. Smoke exposure also induced a dose-dependent injury to tracheobronchial epithelium and lung parenchyma. Taken together these data show that few indices of oxidative stress responded in a dose-dependent manner to graded doses of smoke inhalation, although most of the indices measured in lung were affected by the highest dose of smoke. Additional time course studies are necessary to determine whether these oxidants are a cause or a consequence of the airway and lung injury associated with exposure to wood smoke. PMID- 14751668 TI - Development and standardization of a simple binding assay for the detection of compounds with affinity for the androgen receptor. AB - Concerns have been raised whether natural and man-made chemicals might have the potential of interfering with the endocrine system. Especially interactions with sex hormone receptors are considered as a critical issue. Weak anti-androgenicity has been demonstrated for some environmental pollutants such as p,p'-DDE, and androgenic activity was found in feedlot and pulp mill effluents. In order to be able to screen for compounds with affinity for the androgen receptor (AR), we developed an AR binding assay using a recombinant AR as receptor source and the synthetic androgen methyltrienolone (R 1881) as ligand. Experiments were performed on 96-well microtitre plates. Following method optimization, compounds recently recommended for the validation of assays characterizing AR-mediated effects and those being used for the OECD validation of the Hershberger assay were employed amongst others to standardize the method. The assay readily detected and discriminated compounds with strong and weak affinity for the AR such as natural and synthetic androgens, anti-androgens in therapeutic use, and a variety of chemicals with weak anti-androgenic side effects, whereas in line with previous findings, AR binding properties of dibutylphthalate and its metabolites could not be demonstrated. Detergents interfered with receptor binding, but showed characteristic effects different from that of true AR binding compounds. The assay is simple and sensitive, avoids the use of animals as a receptor source, and should be of value when screening for endocrine-modulating compounds. PMID- 14751669 TI - Therapeutic potential of monoisoamyl and monomethyl esters of meso 2,3 dimercaptosuccinic acid in gallium arsenide intoxicated rats. AB - The dose dependent effects of monoisoamyl and monomethyl esters of meso 2,3 dimercaptosuccinic acid (DMSA) (0.1, 0.3 and 0.5 mmol kg(-1), intraperitoneally (i.p.) once daily for 5 days) to offset the characteristic biochemical, immunological, oxidative stress consequences and DNA damage (based on DNA fragmentation and comet assay) following sub-chronic administration of gallium arsenide and the mobilization of gallium and arsenic were examined. The effects of these chelators alone in normal animals too were examined on above-mentioned variables. Male Wistar rats were exposed to 10 mg kg(-1), GaAs, orally once daily for 12 weeks and were administered DMSA or two of its monoesters (monoisoamyl or monomethyl) for 5 consecutive days. DMSA was used as a positive control. DMSA and its derivatives, when given alone, generally have no adverse effects on various parameters. After 5 days of chelation therapy in GaAs pre-exposed rats, MiADMSA was most effective in the reduction of inhibited blood delta-aminolevulinic acid dehydratase (ALAD) activity and zinc protoporphyrin level while, all three chelators effectively reduced urinary ALA excretion, compared to GaAs alone exposed rats. MiADMSA was also effective, particularly at a dose of 0.3 mmol kg( 1), in enhancing the inhibited hepatic transaminase activities. Parameters indicative of oxidative stress responded less favorably to the chelation therapy, however, three chelators significantly restored the altered immunological variables. MiADMSA was relatively more effective than the other two chelators. GaAs produced significant DNA damage in the liver and kidneys and the chelation treatment had moderate but significant influence in reducing DNA damage. All three chelators significantly reduced arsenic concentration and, however, MiADMSA was more effective than the other two chelators in depleting arsenic concentration from blood and other soft tissues. A dose of 0.3 mmol kg(-1) was found to be relatively better than the other two doses examined. Gallium contents of blood and soft tissues remained uninfluenced by the chelation therapy. Significant loss of copper after MiADMSA administration, however, is of concern and requires further exploration. Additionally, further studies are required for the choice of appropriate dose, duration of treatment and possible toxic/side effects. Keeping in view the promising role of MiADMSA in the treatment of GaAs poisoning, these data will be needed for the registration of this chelating agent as licensed drug for the treatment of gallium arsenide intoxication. PMID- 14751670 TI - Correlation between clinical indicators of lead poisoning and oxidative stress parameters in controls and lead-exposed workers. AB - The present study was undertaken to investigate the involvement of oxidative damage in lead-induced toxicity in humans and to enlighten whether oxidative stress indicators are correlated with the known indices of lead toxicity. For these purposes, selected oxidative stress parameters along with some clinical indices of lead poisoning were determined in blood of battery plant workers and control subjects. Workers had significantly increased erythrocyte malondialdehyde (MDA) levels, catalase and glucose-6-phosphate dehydrogenase (G6PD) activities, and decreased blood glutathione:glutathione disulfide ratio compared to the controls. Increased blood lead concentrations and zinc protoporphyrin (ZPP) levels, and decreased delta-aminolevulinic acid dehydratase (ALAD) activity were used as clinical indices of lead toxicity. Statistically significant correlation between oxidative stress parameters and clinical indices implies that disrupted prooxidant/antioxidant balance might contribute to lead-induced toxicity in erythrocytes. A significant correlation was found between ALAD activity and blood lead levels in human subjects. Similarly significant correlation between ALAD activity and erythrocyte MDA concentrations was shown. Present data indicates that ALAD can serve as a valuable biomarker of oxidative stress in lead-exposed hematological system as well as being a biochemical indicator of lead exposure. PMID- 14751671 TI - Montserrat volcanic ash induces lymph node granuloma and delayed lung inflammation. AB - OBJECTIVES: A substantial amount of Montserrat volcanic ash, containing up to 24% of cristobalite (w/w), a fibrogenic crystalline silica, has been generated since the first documented eruption in 1995. The bioreactivity of the ash and its two major components: cristobalite and anorthite have been studied in vivo for a year following intratracheal instillation into rats. METHODS: The rats (n=5) were instilled with a sterile vehicle solution (0.15 M NaCl) and/or three doses (1.0, 2.5 or 5.0 mg) of each of the dust, and were sacrificed at 13, 25 or 49 weeks post-instillation for quantitative biochemical and histopathological analyses in the lung and lymph nodes. RESULTS: Cristobalite caused inflammation in the lung and granuloma in the hilar lymph nodes associated with significant size augmentation at 13 weeks post-instillation (P<0.05) and cristobalite (5.0 mg) induced fibrosis in the lung at 49 weeks post-exposure. However, the Montserrat volcanic ash caused inflammation in the lung at 49 weeks post-treatment without any fibrogenic response although the ash (5.0 mg) triggered significant lymph node enlargement without significant changes in the lung at 13 weeks post treatment (P<0.05). Dose and time independent responses in the anorthite-exposed lung and lymph nodes suggest that a single instillation of 5.0 mg of poorly soluble mineral dust does not induce any change in the lung or lymph nodes. CONCLUSION: The ash produces inflammatory reactions in lymph nodes at 13 weeks post-instillation in rats. These effects are seen much earlier than any inflammatory reaction in the lung. The onset of the lung inflammation is delayed until 49 weeks post-exposure. Despite the high cristobalite content of the ash, there is no evidence of any lung fibrogenic responses. PMID- 14751672 TI - Silica-exposed mice generate autoantibodies to apoptotic cells. AB - Silica exposure has been associated with development of autoantibodies and systemic autoimmune disease, but mechanisms leading to these events are unknown. This study tested the hypothesis that autoantibodies associated with silica exposure may recognize epitopes on apoptotic macrophages. Serum was obtained from New Zealand mixed (NZM) mice, in which instillation of silica significantly increased production of autoantibodies. Sera were selected that were shown, by indirect immunofluorescence (IIF), to be positive or negative for antinuclear antibodies (ANA) following silica or saline exposure, respectively. Apoptosis was induced in MH-S murine macrophages using silica or cycloheximide. The ability of the autoantibodies to preferentially recognize apoptotic cells was tested using IIF and ELISA. Apoptotic cells, but not live cells, were shown to stain with serum from ANA-positive mice, but not from ANA-negative serum. In addition, binding of antibodies from ANA-positive mice was shown to be significantly greater on cellular lysates from apoptotic cells, but not necrotic or live cell lysates using an ELISA based assay. Finally, inhibition of apoptosis with a broad spectrum caspase inhibitor, Boc-D-FMK, blocked the increased binding by the autoantibodies. These results suggest that autoantibodies from mice with silica exacerbated autoimmune responses recognize specific epitopes on apoptotic macrophages. It is therefore possible that silica-induced apoptosis may exacerbate autoimmune responses by exposing antigenic epitopes to the immune system. PMID- 14751673 TI - Comparison of the Hershberger assay and androgen receptor binding assay of twelve chemicals. AB - We performed the Hershberger assay of 12 chemicals based on the OECD draft protocol. The chemicals tested by the Hershberger assay were phthalic acid di-n hexyl ester, phthalic acid di-n-amyl ester, phthalic acid di-n-propyl ester, diethylstilbestrol, 17beta-estradiol, tamoxifen, 5alpha-dihydrotestosterone, dichlorodiphenyldichloroethane, cyproterone acetate, 6alpha-methyl-17alpha hydroxy-progesterone, atrazine, and spironolactone. Phthalic acid di-n-hexyl ester, phthalic acid di-n-amyl ester, and phthalic acid di-n-propyl ester are phthalates; diethylstilbestrol and 17beta-estradiol are estrogenic chemicals; tamoxifen is partial estrogen receptor antagonist with mainly estrogenic properties; 5alpha-dihydrotestosterone is an androgen derivatives; dichlorodiphenyldichloroethane is a reference androgen antagonistic chemical; cyproterone acetate, 6alpha-methyl-17alpha-hydroxy-progesterone, and spironolactone have an androgenic steroid structure and are known as androgen antagonistic chemicals; and atrazine is a reference endocrine disruptor. We also subjected these chemicals to the receptor binding assay for androgen. A clear androgen agonistic effect was detected in 5alpha-dihydrotestosterone, and an androgen antagonistic effect was observed in five chemicals: cyproterone acetate, spironolactone, 6alpha-methyl-17alpha-hydroxy-progesterone, phthalic acid di-n amyl ester, and dichlorodiphenyldichloroethane. By contrast, diethylstilbestrol, 17beta-estradiol, tamoxifen, 5alpha-dihydrotestosterone, dichlorodiphenyldichloroethane, cyproterone acetate, 6alpha-methyl-17alpha hydroxy-progesterone, and spironolactone were positive in the receptor binding assay for androgen. Three estrogenic chemicals, diethylstilbestrol, 17beta estradiol, and tamoxifen, were negative in the Hershberger assay with receptor binding affinity. On the other hand, the Hershberger assays of three phthalates were performed at the same dosages, and the results showed androgen antagonistic affinity only in the assay of phthalic acid di-n-amyl ester without receptor binding affinity. PMID- 14751674 TI - Preliminary characterisation of an in vitro paradigm for the study of the delayed effects of organophosphorus compounds: hen embryo brain spheroids. AB - Organophosphate induced delayed neuropathy (OPIDN) has been studied extensively but the mechanisms of toxicity remain unclear. It is generally accepted that the inhibition and ageing (dealkylation) of the B-esterase neuropathy target esterase (NTE) is integral to axonal loss. At present, the only way of detecting compounds that induce OPIDN is the hen test, an animal model. In this study, we preliminary validated hen embryo brain spheroids (HEBS) for the study of organophosphate (OP) toxicity. Hen brain spheroids have been characterised previously, although they have never been fully optimised for OP testing. We optimised the levels of acetylcholine esterase (AChE) and neuropathy target esterase by adapting the culture technique and using chemically defined media. Spheroid cultures were maintained for 35 days and viability and enzyme levels were monitored over this time. Levels of AChE and NTE in this system remained stable over the 35 day period. Using transmission electron microscopy, we have shown synaptogenesis within HEBS earlier than previously suggested in spheroid culture. These studies indicate that HEBS may be useful for the study of OP-induced toxicity and that the long-term stability of the cultures makes it an ideal candidate for studying OPIDN. PMID- 14751675 TI - HCH and DDT residues in human placentas in Murcia (Spain). AB - Organochlorinated insecticides are ubiquitous toxicants that are transplacentally transferred from mother to fetus and are reported to produce adverse health effects in pregnant woman and neonates. To investigate hexachlorocyclohexane (HCH) and 1,1,1-trichloro-2,2-bis(4-chlorophenyl)ethane (DDT) exposure in pregnant woman, a total of 102 placentas were analyzed by a gas chromatograph equipped with electron capture detector. Organochlorine residues were found in 69.6% of the samples. Lindane was detected in 24.5%, alpha-HCH in 2.9%, beta-HCH in 6.9%, p,p'-DDE in 44.1%, p,p'-DDT in 14.7% and p,p'-DDD in 10.8% of the samples. The pattern of dispersal by these substances in Murcia is similar to that described by different authors in other countries. Therefore, the past use of DDT and the present employment of lindane seem to be reflected in body tissues of the residents of this area. PMID- 14751676 TI - Foetal rat lung epithelial (FRLE) cells: alterations in cellular homeostasis and gene expression in response to etoposide, hydrogen peroxide and sodium butyrate. AB - Genomics technology offers a way of detecting the effects of a toxin on the expression of many genes in a single experiment. We have previously partially characterised a foetal rat lung epithelial (FRLE) cell line and shown that it is suitable for use in a pneumocytotoxicity screen. In this study, we wanted to ascertain whether we could use alterations in FRLE cell gene expression as a sensitive marker of cell stress. Sodium butyrate and etoposide were shown to arrest FRLE cell cycle at G0/G1 and G2/M phase of the cell cycle, respectively and this was associated with a decrease in the number of cells in culture. Following 24 h of culture both compounds caused a statistically significant increase in the mRNA levels of the cell cycle inhibitory protein, gadd153, whereas p21 was statistically altered by etoposide only. Hydrogen peroxide induced growth arrest at low concentrations (< or =250 microM) following 24 h of culture. We could not detect an increase in apoptosis or in the mRNA levels of the pro-apoptotic protein bax in FRLE cells following culture with hydrogen peroxide or etoposide. Thus, it was possible to correlate cellular perturbations in FRLE cells with alterations in gene expression, demonstrating that these cells are suitable for use in a toxicity screen. PMID- 14751677 TI - Selenium and high dose vitamin E administration protects cisplatin-induced oxidative damage to renal, liver and lens tissues in rats. AB - Cisplatin is one of the most active cytotoxic agents in the treatment of cancer but its clinical use is associated with nephrotoxicity. Several studies suggest that supplementation with antioxidant can influence cisplatin induced nephrotoxicity. In the present study, we investigated the effect of selenium with high dose vitamin E administration on lipid peroxidation (MDA) and scavenging enzyme activity in kidneys, liver and lens of cisplatin-induced toxicity in rats. Forty female Wistar rats were used. They were randomly divided into five groups. The first and second groups were used as control and cisplatin (6 mg/kg BW) intraperitoneally administrated groups. Groups III, IV and V received intraperitoneally five doses of selenium (1.5 mg/kg BW) and a high dose of vitamin E (1000 mg/kg BW) combination before, simultaneously and after with cisplatin, respectively. Glutathione peroxidase (GSH-Px), vitamin E and beta carotene levels in the kidney, lens and liver, vitamin A and reduced glutathione (GSH) levels in the kidney were significantly (P<0.05 to <0.001) lower in the cisplatin group than in the control whereas there was a significant increase in kidney, liver and lens MDA levels in rats treated with cisplatin. The decreased antioxidant enzymes and vitamins and increased MDA levels in the kidney, lens and liver of animals administered with cisplatin were significantly (P<0.05 to <0.001) improved with selenium and a high dose vitamin E injection. In conclusion, this data demonstrates that there is an increase in lipid peroxidation in the kidney, liver and lens of animals administered with cisplatin whereas there is a decrease in antioxidant vitamins and enzymes. However, intraperitoneally injected selenium combined with a high dose of vitamin E seem to produce a significant improvement on antioxidants concentrations in rats treated before, simultaneously and after with cisplatin. The selenium with high dose vitamin E injection may play a role in preventing cisplatin-induced nephropathy and cataract formation in cancer patient. PMID- 14751678 TI - Occupational exposure to styrene: modulation of cytogenetic damage and levels of urinary metabolites of styrene by polymorphisms in genes CYP2E1, EPHX1, GSTM1, GSTT1 and GSTP1. AB - Styrene is widely used in the production of various plastics, synthetic rubber and resins. The aim of this study was to evaluate if individual polymorphisms in xenobiotic metabolizing enzymes, related with the metabolic fate of styrene, could modify individual susceptibility to the possible genotoxic effects of the styrene exposure. Twenty-eight reinforced plastic workers and 28 control subjects were studied. In the selected population the urinary styrene metabolites mandelic (MA) and phenylglyoxylic (PGA) acids were quantified, sister chromatid exchanges (SCE) and micronuclei (MN) were assessed in peripheral lymphocytes and all the subjects were genotyped for GSTM1, GSTT1 (gene deletions), GSTP1 (codon 105 ile==>val), EPHX1 (codons 113 tyr==>his and 139 his==>arg) and CYP2E1 (DraI polymorphism in intron 6). The results obtained showed a significant difference between the levels of SCE, but not in MN levels, in exposed workers as compared with the control group. The GSTP1 and CYP2E1 individual genotypes modulate the baseline levels of SCE that are lower in non-wild type individuals for both polymorphisms. The GSTM1 null individuals with low levels of exposure have significantly higher urinary levels of MA+PGA. The present data seem to suggest that apart from the methodology usually used for monitoring populations occupationally exposed to styrene (urinary metabolites and biomarkers of early biological effects) the analysis of individual genotypes associated with the metabolic fate of styrene should also be carried out in order to evaluate the individual genetic susceptibility of exposed populations. PMID- 14751679 TI - Antiandrogenic activity of extracts of diesel exhaust particles emitted from diesel-engine truck under different engine loads and speeds. AB - To clarify the alteration of androgenic and antiandrogenic activities by diesel engine conditions, we collected diesel exhaust particles (DEP) samples emitted from a diesel-engine truck under different conditions of engine loads and vehicle speeds, and DEP extract (DEPE) samples were prepared from each. The androgenic and antiandrogenic activities of the DEPE samples were examined using a prostate specific antigen (PSA) promoter-luciferase reporter gene assay in PC3/AR human prostate cancer cells. While all DEPE samples did not exhibit androgenic effects, the antiandrogenic effects were enhanced by higher engine load but not by higher vehicle speed. In this study, significant correlations between antiandrogenic and aryl hydrocarbon receptor (AhR) agonistic activities were demonstrated in PC3/AR cells by 16 polycyclic aromatic compounds and beta-naphthoflavone. Yeast two hybrid assay and cytochrome P450 (CYP) 1A1 promoter-luciferase reporter gene assay showed that the antiandrogenic constituents acting as androgen receptor (AR) antagonists and AhR agonists were increased by only the higher engine load. In conclusion, the antiandrogenic effects of DEPE samples were enhanced by a higher engine load which resulted in DEPC samples with elevated AhR agonistic and AR antagonistic activities. PMID- 14751680 TI - Detection of Escherichia coli O157 in bovine meat products in northern Italy. AB - Tests for Escherichia coli and E. coli O157 were carried out on meat samples collected from randomly chosen stores throughout the city of Bologna and suburban areas. The samples consisted of 25 g of loose minced beef, sometimes already shaped into meatballs or hamburgers, some of which were mixed with vegetables. The meat was purchased from retail outlets, open market stalls, and supermarket chains during 25 sampling visits from October 2000 to December 2001. For E. coli detection, Tryptone soya broth (TSB) supplemented with novobiocin and C-EC agar were used. Immunomagnetic separation with SMAC-BCIG-CT agar and chromogenic E. coli O 157 agar, API 20E system and agglutination latex test were used to detect E. coli O157; Vero cell assay and polymerase chain reaction (PCR) were used to assess toxin production and the presence of virulence genes.E. coli were detected in 45 (30.2%) of the 149 samples examined, mainly in the hamburger samples mixed with vegetables and in the loose minced beef. E. coli O157 was found in one sample of hamburger and two samples of hamburger mixed with vegetables (2%) collected from three different butcher's stores between July and October. All the strains of E. coli O157 and most cases of E. coli were found in meat from small retailers. The three strains of E. coli O157 were positive for verocytotoxin production. PCR analysis revealed genes coding for vt2 and one strain possessed the gene for eae A. Chromogenic E. coli O157 agar was found to be more selective and differential, allowing easier identification of suspected colonies with mixed flora and producing less false-positive colonies. PMID- 14751681 TI - Control of Aeromonas on minimally processed vegetables by decontamination with lactic acid, chlorinated water, or thyme essential oil solution. AB - Aeromonas is an opportunistic pathogen, which, although in low numbers, may be present on minimally processed vegetables. Although the intrinsic and extrinsic factors of minimally processed prepacked vegetable mixes are not inhibitory to the growth of Aeromonas species, multiplication to high numbers during processing and storage of naturally contaminated grated carrots, mixed lettuce, and chopped bell peppers was not observed. Aeromonas was shown to be resistant towards chlorination of water, but was susceptible to 1% and 2% lactic acid and 0.5% and 1.0% thyme essential oil treatment, although the latter provoked adverse sensory properties when applied for decontamination of chopped bell peppers. Integration of a decontamination step with 2% lactic acid in the processing line of grated carrots was shown to have the potential to control the overall microbial quality of the grated carrots and was particularly effective towards Aeromonas. PMID- 14751682 TI - Observations on the succession dynamics of lactic acid bacteria populations in chill-stored vacuum-packaged beef. AB - Drip samples were collected at 4-week intervals from 10 vacuum-packaged beef striploins stored for 16 weeks at -1.5 degrees C and assayed for populations of lactic-acid bacteria (LAB), pH and spoilage-causing fermentation products. A total of 15 LAB species were identified using pulsed-field gel electrophoresis and biochemical analysis. A pattern of succession was observed during storage between strains of Carnobacterium, Lactobacillus, Leuconostoc and Pediococcus. Acetic acid production was associated with increasing LAB populations generally and butyric acid production was associated with the development of a particular strain of Leuconostoc. Changes in pH is postulated as a driver of succession. PMID- 14751683 TI - A rapid turbidometric microplate bioassay for accurate quantification of lactic acid bacteria bacteriocins. AB - A 1 day turbidometric microplate bioassay (TMB) was developed for the rapid, accurate and precise quantification of lactic acid bacteria (LAB) bacteriocins (nisin Z and pediocin PA-1). Parameters such as the concentration of the indicator strains and the incubation time were optimized for each bacteriocin. A high correlation coefficient (r(2)=0.992+/-0.004) was obtained for the exponential regression in the nisin Z concentration range of 20-120 ng/ml with 1 x 10(7) CFU indicator strain (Pediococcus acidilactici UL5) and an incubation time of 3 h. Using these parameters, the detection limit was estimated at 80 ng/ml (3.2 IU/ml), compared to 300 ng/ml for the agar diffusion assay (ADA). High precision (<7%) and accuracy (10%) were obtained for all nisin Z concentrations tested. Similar results were obtained with pediocin PA-1 with r(2)=0.993+/-0.005, a precision (8.2%) and an accuracy lower than 15%. PMID- 14751684 TI - Effect of iron limitation on the growth and cytotoxin production of Salmonella choleraesuis SC-5. AB - This study investigated the effect of iron limitation on the growth and cytotoxin production of Salmonella choleraesuis and examined the iron-accruing capability of various Salmonella strains. It was found that the growth of S. choleraesuis SC 5 was retarded by the presence of iron-chelating agents, 2,2'-dipyridyl or ethylenediamine di-o-hydroxyphenylactic acid (EDDA). Addition of 2,2'-dipyridyl to Trypticase soy broth (TSB) resulted in a smaller maximum population of S. choleraesuis noted at the stationary phase, while addition of EDDA in TSB only caused an extended period of lag phase. A significant increase in cytotoxin production was found when S. choleraesuis SC-5 was cultured in iron-limited TSB containing 2,2'-dipyridyl. On the other hand, pre-culture in an iron-limited medium increased the growth of S. choleraesuis SC-5 in mouse serum with or without complement inactivated. Testing with a plate assay method revealed that the ability and efficiency of Salmonella to acquire iron under iron-limited condition varied with strains, and the kinds and dosages of iron-containing compounds present. PMID- 14751685 TI - Prevalence of Arcobacter species in retail meats and antimicrobial susceptibility of the isolates in Japan. AB - A survey was conducted to examine the prevalence of Arcobacter species among meat samples and to investigate the antimicrobial susceptibility of the isolates in Japan. In 1998 and 1999, samples of beef (n=90), pork (n=100) and chicken meat (n=100) were purchased from seven retail shops. Arcobacter species were isolated from 2.2%, 7.0% and 23.0% of beef, pork and chicken meat samples, respectively. The rate of isolations in chicken meats was shown to be significantly higher than those of beef and pork. Species-specific polymerase chain reaction (PCR) demonstrated that the most dominant Arcobacter species was Arcobacter butzleri among the isolates examined. Multiple contaminations with different Arcobacter species were observed in 5% of the chicken samples. Almost all the strains tested showed resistance to vancomycin (100%) and methicillin (97.5%). Strains resistant to cephalothin, sulfamethoxazole-trimethoprim, nalidixic acid and chloramphenicol were detected at the rate of 81.1%, 67.2%, 53.5% and 24.6%, respectively. All Arcobacter strains examined were susceptible to ampicillin, tetracycline, streptomycin and kanamycin. PMID- 14751686 TI - Phenotypic and genetic diversity of Lactococcus lactis and Enterococcus spp. strains isolated from Northern Spain starter-free farmhouse cheeses. AB - To evaluate a previous phenotypic classification of lactococci, 39 presumed lactococcal strains were classified by molecular techniques. The strains were also subjected to several typing techniques to estimate the phenotypic and genetic diversity present in original populations from starter-free farmhouse cheeses. Partial Amplified rDNA Restriction Analysis (partial ARDRA) with either restriction enzyme MboII or HhaI divided these isolates into four distinctive groups. Sequencing of representative amplicons identified 29 isolates as belonging to Lactococcus lactis subsp. lactis (24) and Lactococcus lactis subsp. cremoris (5). The remaining 10 isolates were shown to be Enterococcus durans (8) and Enterococcus faecalis (2), which were misclassified by the traditional tests. Thus, partial ARDRA was successfully used to classify wild Lactococcus-like strains into Lactococcus and Enterococcus species. The technique also allowed differentiation of L. lactis strains at subspecies level. The 29 strains of L. lactis showed five different fermentation profiles, four distinct Random Amplification of Polymorphic DNA (RAPD) profiles, and 14 unrelated profiles by both Restriction Fragment Length Polymorphism analyzed by Pulsed Field Gel Electrophoresis (RFLP-PFGE) and Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis (SDS-PAGE). Using the same techniques, the 10 enterococcal strains showed four fermentation profiles, four RADP, and six by RFLP-PFGE and SDS-PAGE, respectively. Several typing techniques, especially RFLP-PFGE and SDS PAGE, revealed wide phenotypic and genetic variability in both the lactococcal and enterococcal isolates. Two simple, rapid and cheap techniques (partial ARDRA and SDS-PAGE) are proposed as reliable tools for the classification and typing of new lactococcal-like isolates. PMID- 14751687 TI - Emergence of variants with altered survival properties in stationary phase cultures of Campylobacter jejuni. AB - During the stationary phase of Campylobacter jejuni NCTC 11351 viable numbers fluctuate in a characteristic fashion. After reaching the maximum cell count (ca. 2 x 10(9) CFU/ml) in early stationary phase (denoted phase 1), viable numbers subsequently decrease to about 10(6) CFU/ml after 48 h and then increase again to about 10(8) CFU/ml (denoted phase 2) before decreasing once more to a value intermediate between the previous maximum and minimum values. To investigate whether the increase in viable numbers following the initial decline was due to the emergence of a new strain with a growth advantage in stationary phase analogous to the 'GASP' phenotype described in Escherichia coli [Science 259 (1993) 1757], we conducted mixed culture experiments with cells from the original culture and antibiotic-resistant marked organisms isolated from the re-growth phase. In many experiments of this type, strains isolated from phase 2 failed to out-compete the original strain and we have thus been unable to demonstrate a convincing GASP phenotype. However, strains isolated from phase 2 showed a much lower rate of viability loss in early stationary phase and a small increase in resistance to aeration, peroxide challenge and heat, indicating that the emergent strain was different from the parent. These results support the view that dynamic population changes occur during the stationary phase of C. jejuni that may play a role in the survival of this organism. PMID- 14751688 TI - Studies to determine the critical control points in pork slaughter hazard analysis and critical control point systems. AB - Aerobic mesophilic counts (AMC), coliform (CC) and coliform resuscitation counts (CRCs) were obtained by swabbing 50 cm(2) areas at three sites (ham, belly and neck) on pig carcasses, after each of seven stages of the slaughter/dressing process (bleeding, scalding, dehairing, singeing, polishing, evisceration and chilling). In most cases, there were no statistical differences (P>0.05) among the counts derived by these three methods. Reductions in counts at individual sites were observed after scalding (3.5 log(10) cfu cm(-2)), and singeing (2.5 log(10) cfu cm(-2)). Increases in counts at individual sites were observed after dehairing (2.0 log(10) cfu cm(-2)) and polishing (1.5 log(10) cfu cm(-2)). The incidence of Salmonella on pig carcasses was also obtained by swabbing the outside surfaces of 100 half carcasses. Information on the incidence of Salmonella in scald tank water (108 samples) was also investigated. Carcass swabs and scald tank water were examined for the presence of Salmonella using standard enrichment methods. Salmonella were detected on 31% of carcasses immediately after bleeding, 7% of carcasses immediately after dehairing and evisceration, and 1% of carcasses immediately after scalding. Serovars included Salmonella Typhimurium, Salmonella Hadar, Salmonella Infantis and Salmonella Derby. No Salmonella were recovered from samples of scald tank water. The impact of pig slaughter/dressing processes on carcass microbiology and their potential use as critical control points (CCPs) during pork production are discussed. PMID- 14751689 TI - Characterization of Listeria monocytogenes and Listeria innocua from a vegetable processing plant by RAPD and REA. AB - The incidence of Listeria monocytogenes in a vegetable processing plant was investigated over a 23-month period. Frozen ready-to-eat vegetable samples, well as the plant environment, were sampled. The molecular subtyping techniques, Random Amplified Polymorphic DNA (RAPD) and Restriction Endonuclease Analyses (REA), were performed to help investigate the origin and routes of Listeria dissemination. The low and sporadic incidence of L. monocytogenes made it impossible to establish an epidemiological sequence in the processing plant, though a case of cross-contamination between tomato and ratatouille was detected. Listeria innocua subtyping, however, allowed us to determine the prevalence of several strains in vegetables, and their presence on machinery samples suggested the possibility of cross-contamination during processing. The low incidence of L. monocytogenes indicated that the risk of listeriosis transmission by vegetable consumption is low. On the other hand, the isolation of the same strain of L. innocua in several surveys pointed out the risk of colonisation on surfaces and machinery. The persistence of Listeria spp. is a cause for concern as can lead to future contamination of vegetables processed in the plant and to a possible increased risk for health. Therefore, periodic controls for the presence of Listeria spp. and a further review of the cleaning and disinfection procedures used in frozen vegetable plants are recommended. PMID- 14751690 TI - Occurrence of Listeria monocytogenes in fresh and processed foods in Navarra (Spain). AB - The presence of Listeria spp. was investigated in a total of 3685 food samples obtained from different industries and markets of Northern Spain in the last 4 years. The samples analyzed include fresh raw products (meat, milk and poultry) and treated products (cooked and cured meats, frozen vegetables and smoked salmon). Occurrence of Listeria spp. varied from 8.1% in soft cheese to 76.3% in raw poultry samples. The highest incidence of L. monocytogenes also occurred in raw poultry (36.1% positive samples). Despite this high incidence of contamination, these kinds of products carry a low risk of listeriosis transmission because of the heat treatment prior to consumption. On the other hand, the ready-to-eat products (RTE) tested in this study showed incidences that could pose serious health problems, taking into account that the storage conditions may allow for rapid growth of the pathogen. It was also found that up to 75.5% of the L. monocytogenes strains isolated in this study belonged to serogroup 1, mainly serotype 1/2a, while the clinical cases observed in Navarra in the same period of time belonged mainly to serotype 4b/4bx. PMID- 14751691 TI - Binding of Bifidobacterium bifidum and Lactobacillus reuteri to the carbohydrate moieties of intestinal glycolipids recognized by peanut agglutinin. AB - We examined binding of Bifidobacterium bifidum and Lactobacillus reuteri to the carbohydrate moieties of glycolipids extracted from human enterocyte-like Caco-2 cells in this study. In binding assays to reference glycolipids of different carbohydrate compositions, B. bifidum EB102 bound strongly to gangliotetraosylceramide (asialo-GM1) and less strongly to gangliotriaosylceramide (asialo-GM2), lactosylceramide and sulfatide. The binding profile of B. bifidum EB102 was almost identical to that of L. reuteri JCM1081 described previously [Lett. Appl. Microbiol. 27 (1998) 130]. When we examined binding to neutral glycolipids extracted from Caco-2 cells, the binding profiles of B. bifidum EB102 and L. reuteri JCM1081 were very similar to that shown by peanut agglutinin (PNA). Binding of both strains to periodate-treated intestinal glycolipids was completely abolished, suggesting that the bacterial cells bind to carbohydrate moieties of the glycolipids. Furthermore, B. bifidum EB102 was found to express multiple glycolipid-binding proteinaceaous components on the cell surface. These results strongly suggested involvement of cell-surface proteinaceous components of B. bifidum in binding to the carbohydrate moieties of intestinal glycolipids recognized by PNA. Binding ability of B. bifidum and L. reuteri to intestinal glycolipids may play a crucial role for colonization on the mucosal surface of the intestine. PMID- 14751692 TI - Avian influenza: the threat looms. PMID- 14751693 TI - Should prenatal diagnostic testing be offered to all pregnant women on economic grounds? PMID- 14751694 TI - Burn scars: a new cause of vitamin D deficiency? PMID- 14751695 TI - Retrovirus infection as a trigger for primary biliary cirrhosis? PMID- 14751697 TI - "Personal account": a new feature from the patient's perspective. PMID- 14751698 TI - Comparison of outcomes in cancer patients treated within and outside clinical trials: conceptual framework and structured review. AB - BACKGROUND: Many oncologists believe that patients with cancer who enroll in clinical trials have better outcomes than those who do not enroll. We aimed to assess the empirical evidence that such a trial effect exists. METHODS: We developed a conceptual framework for comparison of trial and non-trial patients. We then did a comprehensive literature search to identify studies that compared outcomes between these groups. We critically evaluated these studies to assess whether they provide valid and generalizable support for a trial effect. FINDINGS: We identified 26 comparisons, from 24 published articles, of outcomes among cancer patients enrolled and not enrolled in clinical trials. 21 comparisons used retrospective cohort designs. 14 comparisons provided some evidence that patients enrolled in trials have improved outcomes. However, strategies to control for potential confounding factors were inconsistent and frequently inadequate. Only eight comparisons restricted non-trial patients to those meeting trial eligibility criteria. Of these, three noted better outcomes in trial patients than in non-trial patients. Children with cancer, patients with haematological malignant disease, and patients treated before 1986 were disproportionately represented in positive studies. INTERPRETATION: Despite widespread belief that enrollment in clinical trials leads to improved outcomes in patients with cancer, there are insufficient data to conclude that such a trial effect exists. Until such data are available, patients with cancer should be encouraged to enroll in clinical trials on the basis of trials' unquestioned role in improving treatment for future patients. PMID- 14751699 TI - Doubling the dose of inhaled corticosteroid to prevent asthma exacerbations: randomised controlled trial. AB - BACKGROUND: Asthma self-management plans that include doubling the dose of inhaled corticosteroid when the condition deteriorates improve asthma control. Whether doubling the dose of corticosteroid in isolation is effective is unknown. We undertook a randomised controlled trial to investigate the effects of doubling the dose of inhaled corticosteriods when asthma deteriorates. METHODS: 390 individuals with asthma who were at risk of an exacerbation monitored their morning peak flow and asthma symptoms for up to 12 months. When peak flow or symptoms started to deteriorate, participants added an active or placebo corticosteroid inhaler to their usual corticosteroid for 14 days to produce a doubling or no change in dose. The primary outcome was the number of individuals starting oral prednisolone in each group. FINDINGS: During 12 months, 207 (53%) started their study inhaler and 46 (12%) started prednisolone--22 (11%) of 192 and 24 (12%) of 198 in the active and placebo groups, respectively. The risk ratio for starting prednisolone was therefore 0.95 (95% CI 0.55-1.64, p=0.8). INTERPRETATION: We recorded little evidence to support the widely recommended intervention of doubling the dose of inhaled corticosteroid when asthma control starts to deteriorate. PMID- 14751700 TI - Cost utility of prenatal diagnosis and the risk-based threshold. AB - BACKGROUND: Prenatal testing guidelines recommend offering amniocentesis or chorionic villus sampling to women aged 35 years or older, or who have been found by screening to be at a similarly high risk of giving birth to an infant with Down's syndrome or another chromosomal abnormality. This threshold was chosen, in part, because 35 was the approximate age at which amniocentesis was cost beneficial when testing guidelines were developed in the USA in the 1970s. We aimed to assess the economic validity of thresholds based on age or risk for offering invasive prenatal diagnosis. METHODS: We did a cost-utility analysis of chorionic villus sampling and amniocentesis versus no invasive testing using data from randomised trials, case registries, and a utility assessment of 534 diverse pregnant women aged 16-47 years. FINDINGS: In the USA, compared with no diagnostic testing, amniocentesis costs less than US15000 dollars per quality adjusted life year gained for women of all ages and risk levels. The results do not depend on maternal age or risk of Down's syndrome-affected birth. The cost utility ratio for any individual woman depends on her preferences for reassurance about the chromosomal status of her fetus, and, to a lesser extent, for miscarriage. INTERPRETATION: Prenatal diagnostic testing can be cost effective at any age or risk level. Current guidelines should be changed to offer testing to all pregnant women, not just those whose risk of carrying an affected fetus exceeds a specified threshold. PMID- 14751701 TI - Variant surface antigen-specific IgG and protection against clinical consequences of pregnancy-associated Plasmodium falciparum malaria. AB - BACKGROUND: Pregnancy-associated malaria caused by Plasmodium falciparum adherence to chondroitin sulfate A in the placental intervillous space is a major cause of low birthweight and maternal anaemia in areas of endemic P falciparum transmission. Adhesion-blocking antibodies that specifically recognise parasite encoded variant surface antigens (VSA) are associated with resistance to pregnancy-associated malaria. We looked for a possible relation between VSA specific antibody concentrations, placental infection, and protection from low birthweight and maternal anaemia. METHODS: We used flow cytometry to measure VSA specific IgG concentrations in plasma samples taken during child birth from 477 Kenyan women selected from a cohort of 910 women on the basis of HIV-1 status, gravidity, and placental histology. We measured VSA expressed by one placental P falciparum isolate and two isolates selected or not selected for chondroitin sulfate A adhesiveness in-vitro. FINDINGS: Concentrations of plasma IgG specific for VSA, expressed by chondroitin sulfate A-adhering parasites (VSA in pregnancy associated malaria or vsa-pam), increased with gravidity and were associated with placental histological findings. Women with chronic pregnancy-associated malaria and low or absent VSA-PAM-specific IgG had lower haemoglobin values (reduced by 17 g/L; 95% CI 8.1-25.2) and delivered smaller babies (birthweight reduced by 0.26 kg; 0.10-0.55) than did corresponding women with high VSA-PAM-specific IgG. No such relation was shown for concentrations of IgG with specificity for non pregnancy-associated malaria VSA. INTERPRETATION: VSA-PAM-specific IgG protects against low birthweight and maternal anaemia. Our data indicate an important mechanism of clinical protection against malaria and raise hope for the clinical effectiveness of a potential VSA-based vaccine against pregnancy-associated malaria. PMID- 14751702 TI - Genetics in reverse. PMID- 14751703 TI - Synthesis of vitamin D in skin after burns. AB - Severe burn injury is associated with vitamin D deficiency, low bone turnover, and abnormalities in calcium homoeostasis. Patients do not routinely receive vitamin D supplementation and sun exposure is currently not controlled. By analysis of skin biopsy samples for vitamin D3 precursors after exposure to ultraviolet B light we found that the conversion of 7-dehydrocholesterol to previtamin D3 was reduced in children a mean of 14 months after the burn. Low serum 25-hydroxyvitamin D concentrations were also found. We conclude that vitamin D supplementation is necessary after burn injury. PMID- 14751704 TI - Group B streptococcal disease in UK and Irish infants younger than 90 days. AB - The incidence, morbidity, and mortality of group B streptococcal disease in the UK and Republic of Ireland are largely unknown. Between Feb 1, 2000, and Feb 28, 2001, we identified cases of invasive group B streptococcal disease in infants younger than 90 days through surveillance involving paediatricians, microbiologists, and parents. 568 cases were identified, equivalent to a total incidence of 0.72 per 1000 live-births (95% CI 0.66-0.78); the incidence for early-onset disease (n=377) was 0.48 per 1000 (0.43-0.53), and for late-onset disease (n=191) was 0.24 per 1000 (0.21-0.28). Risk factors were identifiable for 218 (58%) cases of early-onset disease. 53 infants died (overall 9.7%). We have established the minimum current burden of group B streptococcal disease in UK and Irish infants. This information will assist in the formulation of guidelines for prevention of this disease. PMID- 14751705 TI - Postpartum psychiatric disorders. AB - This review summarises the psychiatry of the puerperium, in the light of publications during the past 5 years. A wide variety of disorders are seen. Recognition of disorders of the mother-infant relationship is important, because these have pernicious long-term effects but generally respond to treatment. Psychoses complicate about one in 1000 deliveries. The most common is related to manic depression, in which neuroleptic drugs should be used with caution. Post traumatic stress disorder, obsessions of child harm, and a range of anxiety disorders all require specific psychological treatments. Postpartum depression necessitates thorough exploration. Cessation of breastfeeding is not necessary, because most antidepressant drugs seem not to affect the infant. Controlled trials have shown the benefit of involving the child's father in therapy and of interventions promoting interaction between mother and infant. Owing to its complexity, multidisciplinary specialist teams have an important place in postpartum psychiatry. PMID- 14751706 TI - Motherhood--a potential killer. PMID- 14751707 TI - Communicating sad, bad, and difficult news in medicine. AB - In every medical specialty bad, sad, and difficult information must be given to patients and their families. An insensitive approach increases the distress of recipients of bad news, may exert a lasting impact on their ability to adapt and adjust, and can lead to anger and an increased risk of litigation. Many doctors also find these interactions stressful, and in the absence of much effective training they may adopt inappropriate ways of delivering bad news and coping with the emotional fall-out. Recognition of these difficulties has led to many initiatives, ranging from increased communication skills training to the development of guidelines and protocols. We review some of the research on the impact that giving sad, bad, and difficult news has on doctors and patients, and assess whether interventions are helping. We focus mainly on difficulties encountered involving parents in an obstetric or paediatric setting, people in acute trauma situations such as accident and emergency departments, and patients with cancer. PMID- 14751708 TI - Scaling-up treatment for HIV/AIDS: lessons learned from multidrug-resistant tuberculosis. AB - The UN has launched an initiative to place 3 million people in developing countries on antiretroviral AIDS treatment by end 2005 (the 3 by 5 target). Lessons for HIV/AIDS treatment scale-up emerge from recent experience with multidrug-resistant tuberculosis. Expansion of treatment for multidrug-resistant tuberculosis through the multipartner mechanism known as the Green Light Committee (GLC) has enabled gains in areas relevant to 3 by 5, including policy development, drug procurement, rational use of drugs, and the strengthening of health systems. The successes of the GLC and the obstacles it has encountered provide insights for building sustainable HIV/AIDS treatment programmes. PMID- 14751711 TI - Rosuvastatin-warfarin drug interaction. PMID- 14751710 TI - Bifidobacterium lipoteichoic acid and false ELISA reactivity in aspergillus antigen detection. AB - A major difficulty with the detection of circulating galactomannan, a cell-wall polysaccharide released by Aspergillus sp during growth, in the serodiagnosis of invasive aspergillosis is the occurrence of false-positive ELISA results, especially in neonates and infants. On the basis of molecule similarity, we postulate that a lipoteichoic acid of Bifidobacterium sp can act as epitope for the monoclonal antibody used in the ELISA. The neonatal gut is heavily colonised with Bifidobacterium sp and these bacteria or their lipoteichoic acid might cause ELISA reactivity with serum after translocation because of immaturity of the intestinal mucosa. If our hypothesis is correct, we might find a method to discriminate between false-positive and true-positive ELISA results and thereby prevent unnecessary pre-emptive treatment of patients. PMID- 14751712 TI - Politics in medical journals. PMID- 14751714 TI - What are health and human rights? PMID- 14751715 TI - Quality of medical education in Mexico. PMID- 14751716 TI - Oral prednisolone for viral wheeze in young children. PMID- 14751718 TI - Effects of different blood-pressure-lowering regimens on major cardiac events. PMID- 14751720 TI - Confusion over benefits of hormone replacement therapy. PMID- 14751721 TI - Effects of different blood-pressure-lowering regimens on major cardiac events. PMID- 14751722 TI - If she were your daughter, what would you do? PMID- 14751727 TI - Food fads. PMID- 14751728 TI - Citizen Copito. PMID- 14751729 TI - Measurement of Poisson's ratio of dental composite restorative materials. AB - The aim of this study was to determine the Poisson ratio of resin-based dental composites using a static tensile test method. Materials used in this investigation were from the same manufacturer (3M ESPE) and included microfill (A110), minifill (Z100 and Filtek Z250), polyacid-modified (F2000), and flowable (Filtek Flowable [FF]) composites. The Poisson ratio of the materials were determined after 1 week conditioning in water at 37 degrees C. The tensile test was performed with using a uniaxial testing system at crosshead speed of 0.5 mm/min. Data was analysed using one-way ANOVA/post-hoc Scheffe's test and Pearson's correlation test at significance level of 0.05. Mean Poisson's ratio (n=8) ranged from 0.302 to 0.393. The Poisson ratio of FF was significantly higher than all other composites evaluated, and the Poisson ratio of A110 was higher than Z100, Z250 and F2000. The Poisson ratio is higher for materials with lower filler volume fraction. PMID- 14751730 TI - Controlled degradation of hydrogels using multi-functional cross-linking molecules. AB - Hydrogels, chemically cross-linked or physically entangled, have found a number of applications as novel delivery vehicles of drugs and cells. However, the narrow ranges of degradation rates and mechanical strength currently available from many hydrogels limits their applications. We have hypothesized that utilization of multi-functional cross-linking molecules to form hydrogels could provide a wider range and tighter control over the degradation rates and mechanical stiffness of gels than bi-functional cross-linking molecules. To address the possibility, we isolated alpha-L-guluronate residues of sodium alginate, and oxidized them to prepare poly(aldehyde guluronate) (PAG). Hydrogels were formed with either poly(acrylamide-co-hydrazide) (PAH) as a multi-functional cross-linking molecule or adipic acid dihydrazide (AAD) as a bi-functional cross linking molecule. The initial properties and degradation behavior of both PAG gel types were monitored. PAG/PAH hydrogels showed higher mechanical stiffness before degradation and degraded more slowly than PAG/AAD gels, at the same concentration of cross-linking functional groups. The enhanced mechanical stiffness and prolonged degradation behavior could be attributed to the multiple attachment points of PAH in the gel at the same concentration of functional groups. This approach to regulating gel properties with multifunctional cross-linking molecules could be broadly used in hydrogels. PMID- 14751731 TI - Influence of polymer membrane porosity on C3A hepatoblastoma cell adhesive interaction and function. AB - The effect of the porosity of acrylonitrile-N-vinylpyrrolidone copolymer membranes on human C3A hepatoblastoma cell adhesive interaction and functioning is investigated on four membranes with an average pore size ranging between 6 and 12 nm. Adhesion of C3A cells was quantified and characterized by studying overall cell morphology and focal adhesion formation. Cell-cell interactions were characterized by E-cadherin expression and organization. Cell growth, fibronectin synthesis and cytochrome P450 activity were estimated as criteria of functional cell activity. The results suggest that membrane porosity influences the initial cell-surface interactions since an increasing pore size augmented cell adhesion and aggregate formation. Cell growth after 7 d was diminished on membranes with an average pore size of 12 nm. The activity of P450 measured by 7-ethoxycoumarin conversion at day 7 was influenced by membrane topography representing a clear optimum in the range of 7-10 nm pore size. These results indicate that membrane porosity is a determinant for the function of hepatocytes in extracorporal liver assist devices. PMID- 14751732 TI - A novel electrically conductive and biodegradable composite made of polypyrrole nanoparticles and polylactide. AB - A novel electrically conductive biodegradable composite material made of polypyrrole (PPy) nanoparticles and poly(d,l-lactide) (PDLLA) was prepared by emulsion polymerization of pyrrole in a PDLLA solution, followed by precipitation. The composite was characterized by scanning electron microscopy and X-ray photoelectron spectroscopy. The electrical stability of the composite containing 5 wt% PPy was investigated in a cell culture environment for 1000 h with 100 mV DC applied voltage. Fibroblasts were cultured on the composite membranes and were stimulated with various DC currents. The PPy particles formed aggregations and constituted microdomains and networks embedded in the PDLLA. With the 1-17% increase in the PPy content, the conductivity of the composite increased by six orders of magnitude. The surface resistivity of the PPy/PDLLA membrane with 3% PPy was as low as 1x10(3) Omega/square. The electrical stability was significantly better in the PPy/PDLLA composite than in the PPy-coated polyester fabrics. For the composite with 5% PPy, the test membrane retained 80% and 42% of the initial conductivity in 100 and 400 h, respectively, following the addition of the MEM solution, compared to 5% and 0.1% for the PPy-coated polyester fabrics. Under 100 mV, a composite membrane 3.0x2.5x0.03cm3 in size and containing 5% PPy sustained a biologically meaningful electrical conductivity in a typical cell culture environment for 1000 h. The growth of fibroblasts was up regulated under the stimulation of medium range intensity of DC current. PMID- 14751733 TI - Processing and properties of porous poly(L-lactide)/bioactive glass composites. AB - Porous poly(L-lactide)/bioactive glass (PLLA/BG) composites were prepared by phase separation of polymer solutions containing bioactive glass particles (average particle size: 1.5 microm). The composite microstructures consist of a porous PLLA matrix with glass particles distributed homogeneously throughout. Large pores (>100 microm) are present in a network of smaller (<10 microm) interconnected pores. The porous microstructure of the composites was not significantly influenced by glass content (9 or 29 vol%), but silane pretreatment of the glass resulted in better glass incorporation in the matrix. Mechanical tests showed that an increase in glass content increased the elastic modulus of the composites, but decreased their tensile strength and break strain. Silane pretreatment enhanced the increase in modulus and prevented the decrease in tensile strength with increasing glass content. Composites soaked in simulated body fluid (SBF) at body temperature formed bone-like apatite inside and on their surfaces. The silane pretreatment of glass particles delayed the in vitro apatite formation. This bone-like apatite formation demonstrates the composites' potential for integration with bone. PMID- 14751734 TI - Gap junction communication between cells aggregated on a cellulose-coated polystyrene: influence of connexin 43 phosphorylation. AB - The appropriate functioning of tissues and organ systems depends on intercellular communication such as gap junctions formed by connexin (Cx) protein channels between adjacent cells. We have previously shown that Swiss 3T3 cells aggregated on hydrophilic cellulose substratum Cuprophan (CU) establish short linear gap junctions composed of Cx 43 in cell surface plaques. This phenomenon seems to depend on the high intracellular cyclic AMP (cAMP) concentration triggered by attachment of the cells to CU. We have now used a cellulose-coated polystyrene inducing the same cell behaviour to analyse the gap junction communication between aggregated cells. The transfer of the dye Lucifer Yellow (LY) between cells showed that cells aggregated on cellulose substratum rapidly (within 90 min) establish functional gap junctions. Inhibitors of cAMP protein kinase (PKI) or protein kinase C (GF109203X) both inhibited the diffusion of LY between neighbouring cells. Western blot analysis showed that this change in permeability was correlated with a decrease in Cx 43 phosphorylation. Thus, cellulose substrata seem to induce cell-cell communication through Cx 43 phosphorylation modulated by PKA and PKC. To understand the mechanisms by which a substratum regulates gap junctional communication is critically important for the emerging fields of tissue engineering and biohybrid devices. PMID- 14751735 TI - Phenotypic expression of bone-related genes in osteoblasts grown on calcium phosphate ceramics with different phase compositions. AB - Calcium phosphate ceramics with different hydroxyapatite (HA) and tricalcium phosphate (TCP) ratios have different chemical properties. Does the difference in phase composition affect osteoblast behavior? In this study, osteoblasts were cultured on 4 kinds of calcium phosphate ceramics, i.e. pure (HA), HT1 (HA/TCP, 70/30), HT2 (HA/TCP, 35/65), and pure TCP. Cell proliferation of SaOS-2 cells together with bone-related genes' mRNA expression and protein production in osteoblasts cultured on different calcium phosphate ceramics were detected at different time points. Data suggested that cell proliferation rate on TCP ceramics was lower than that on the other substrates tested. Generally, mRNA expressions for osteonectin and osteocalcin were similar among the four kinds of ceramics in most circumstances, whereas at six days, alkaline phosphatase mRNA expression was higher on HA and HT1 surfaces than on the other two materials. Collagen I mRNA expression was also affected by the phase composition of substrates. Osteocalcin and bone sialoprotein production in SaOS-2 cells was very similar no matter which ceramic surface the cells were grown upon. This study revealed that calcium phosphate ceramics substrate could support osteoblast growth and bone-related gene expression and its gene expression pattern explained the basis of the biocompatibility and bioactivity for calcium phosphate ceramics. PMID- 14751736 TI - MMP-2, MMP-9 and their inhibitors TIMP-2 and TIMP-1 production by human monocytes in vitro in the presence of different forms of hydroxyapatite particles. AB - After calcium-phosphates biomaterials based implantation like hydroxyapatite (HA) coating, particles are released in the periprosthetic tissues. Wear-debris induced fibrous membranes contain macrophage subsets that can produce metalloproteinases (MMPs), which are considered to be key enzymes in extra cellular matrix turnover. Tissue inhibitors of metalloproteinases (TIMPs) are important regulator of MMPs activity. Interleukin-1 mainly produced by monocytes can also regulate MMPs production. In the present work, we have evaluated the effect of HA particles characteristics (size, shape and sintering temperature) on the MMP-2, -9 and their respective inhibitors TIMP-2, -1 production. Our results demonstrate that sintering temperature (that modify crystal size and surface area) have little effect on MMPs and TIMPs production. Non-phagocytable particles induced more MMP-9, although phagocytable particles induced more IL-1beta release. The shape of the particles was the most important factor since needle shaped particles induced the most significant up-regulated expression of MMPs and IL-1beta. PMID- 14751737 TI - Formation and growth of calcium phosphate on the surface of oxidized Ti-29Nb-13Ta 4.6Zr alloy. AB - The bioconductivity of a new biomedical titanium alloy Ti-29Nb-13Ta-4.6Zr achieved by a combination of surface oxidation and alkali treatment is reported in this paper. Oxidation treatment at 400 degrees C for 24 h was found to result in the formation of a hard layer on the surface of the alloy. Immersion in a protein-free simulated body fluid and fast calcification solution led to the growth of calcium phosphate (Ca-P) phase on the oxidized and alkali-treated alloy, and the new bioconductive surface was still harder than the substrate. The surface processes during various treatment and immersion processes were investigated in detail, and the morphology of the calcium phosphate crystals was shown to be determined by the concentrations of Ca and P in the solution. PMID- 14751738 TI - Hydroxyapatite coating on titanium substrate with titania buffer layer processed by sol-gel method. AB - Hydroxyapatite (HA) was coated onto a titanium (Ti) substrate with the insertion of a titania (TiO2) buffer layer by the sol-gel method. The HA layer was employed to enhance the bioactivity and osteoconductivity of the Ti substrate, and the TiO2 buffer layer was inserted to improve the bonding strength between the HA layer and Ti substrate, as well as to prevent the corrosion of the Ti substrate. The HA layer coated over the TiO2 showed a typical apatite phase at 400 degrees C and the phase intensity increased above 450 degrees C. The sol-gel derived HA and TiO2 films, with thicknesses of approximately 800 and 200 nm, respectively, adhered tightly to each other and to the Ti substrate. The bonding strength of the HA/TiO2 double layer coating on Ti was markedly improved when compared to that of the HA single coating on Ti. The highest strength of the double layer coating was 55 MPa after heat treatment at 500 degrees C. The improvement in bonding strength with the insertion of TiO2 was attributed to the resulting enhanced chemical affinity of TiO2 toward the HA layer, as well as toward the Ti substrate. Human osteoblast-like cells, cultured on the HA/TiO2 coating surface, proliferated in a similar manner to those on the TiO2 single coating and on the pure Ti surfaces. However, the alkaline phosphatase activity of the cells on the HA/TiO2 double layer was expressed to a higher degree than that on the TiO2 single coating and pure Ti surfaces. The corrosion resistance of Ti was improved by the presence of the TiO2 coating, as confirmed by a potentiodynamic polarization test. PMID- 14751739 TI - Biocompatible nanocrystalline octacalcium phosphate thin films obtained by pulsed laser deposition. AB - We extended for the first time pulsed laser ablation to the deposition of octacalcium phosphate Ca8H2(PO4)6.5H2O (OCP) thin films. The depositions were performed with a pulsed UV laser source (lambda=248 nm, tau> or =20 ns) in a flux of hot water vapors. The targets were sintered from crystalline OCP powder and the laser ablation fluence was set at values of 1.5-2 J/cm2. During depositions the collectors, Si or Ti substrates, were maintained at a constant temperature within the range 20-200 degrees C. The resulting structures were submitted to heat treatment in hot water vapors for up to 6 h. The best results were obtained at a substrate temperature of 150 degrees C during both deposition and post deposition treatment. High-resolution electron microscopy and XRD at grazing incidence indicated that the coatings obtained were made of nanocrystalline OCP. Cross-section TEM investigations showed that the coatings contained droplets stacked on Ti substrates as well as distributed across the entire thickness of the arborescence-like structure layers. The results of WST-1 assay, cell adherence, DNA replication, and caspase-1 activity confirmed the good biocompatibility of the coatings. PMID- 14751740 TI - Osteoblast response to bioactive glasses in vitro correlates with inorganic phosphate content. AB - Inorganic phosphate (Pi) is a physiological regulator of osteoblasts and chondrocytes, suggesting that phosphate may contribute to the biological response of these cells to bioactive glasses like Bioglass 45S5, which is composed of 45% SiO2, 24.5% CaO, 24.5% Na2O, and 6% P2O5. We investigated the effect of varying the Pi content of bioactive glass disks (0%, 3%, 6% and 12% P2O5) using human osteoblast-like MG63 cells as the model. Cell number on 6% Pi disks was comparable to cultures on tissue culture plastic, but was reduced at higher and lower Pi concentrations. Alkaline phosphatase specific activity of isolated cells and cell layer lysates, as well as PGE2, TGF-beta1 and NO levels in conditioned media, were elevated in cultures grown on bioactive glass and varied with the Pi content. The greatest effects were observed in cultures grown on disks with the lowest Pi concentrations. Thus, growth on the bioactive glasses enhances cell function in comparison with tissue culture plastic and lower Pi content favors osteoblast differentiation. PMID- 14751741 TI - Inflammatory responses and cell adhesion to self-assembled monolayers of alkanethiolates on gold. AB - The acute inflammatory response and the adhesion of cells to self-assembled monolayers (SAMs) of well-defined surface chemistry was studied in vivo using a rodent air-pouch model of inflammation. SAMs with three different terminal functional groups (OH, COOH and CH3) were implanted in subcutaneous air pouches induced in BALB/c mice. After 24 h, inflammatory cells were recovered from the air pouches and the implants were removed and prepared for observation by scanning electron microscopy (SEM). The implants coated with OH and CH3, were found to cause the highest recruitment of inflammatory cells into the subcutaneous pouches. Polymorphonuclear neutrophils (PMNs) leukocytes predominated over mononuclear cells in inflammatory exudates of SAMs-coated implants, the opposite being found in uncoated implants (controls). CH3-coated implants induced the highest number of inflammatory cells and also the largest percentage of PMNs seen in the subcutaneous pouches. Control and OH-covered implants presented the higher densities of attached inflammatory cells detected by SEM. In contrast, the CH3-coated implants showed a very low density of cells adherent to the implant surface. We conclude that the chemical nature and the degree of hydrophobicity of the surface of implants modulate both the local acute inflammatory reaction and the adhesion of leukocytes. PMID- 14751742 TI - Physiological strains remodel extracellular matrix and cell-cell adhesion in osteoblastic cells cultured on alumina-coated titanium alloy. AB - The effects of mechanical strains on cellular activities were assessed in an in vitro model using human osteoblastic MG-63 cells grown on titanium alloy discs coated with porous alumina and exposed to chronic intermittent loading. Strain was applied with a Dynacell device for three 15-min sequences per day for several days with a magnitude of 600 microepsilon strain and a frequency of 0.25 Hz. We have previously demonstrated that this regimen increased alkaline phosphatase activity in confluent cultures on ceramic coated titanium (alumina and hydroxyapatite) (Biomaterials 24 (2003) 3139). In this study, we analysed the production of bone matrix proteins. Osteocalcin secretion quantified by ELISA between day 5 and 11 was not affected by mechanical strain. Strain had even no quantifiable effect on collagen production from day 1 to 5 as measured by carboxy terminal collagen type I propeptide release. On the other hand, stress stimulation resulted in increased expression of fibronectin (FN) measured by Western blot after 1 day stretching. This upregulation of FN production was followed by reorganisation of the FN network after 5 days stretching observed by immunostaining. The receptors for collagen and FN, alpha2beta1, alpha5beta1 and beta1 integrins were not quantitatively affected by the strains as measured by flow cytometry. A modification of cell morphology was seen after 5 days of loading that appeared to increase cell spreading, implying consequences on intercellular contacts. For this reason, N, C11 and E-adherins were examined. We noted a selective effect characterised by increased expression of N-cadherin using both RT-PCR and Western blot analyses. We concluded that reinforcement of cell-cell adhesion and remodelling of the FN network are important adaptive responses to physiological strains for human osteoblasts grown on alumina-coated biomaterials. PMID- 14751743 TI - Characterization of CO3Ap-collagen sponges using X-ray high-resolution microtomography. AB - For reconstruction and regeneration of hard tissues, scaffold biomaterials with large size pores and high porosity are important, in addition to their roles as supporting frames. To develop a new biodegradable scaffold biomaterial, CO3Ap, which has crystallinity and a chemical composition similar to bone, was synthesized at pH 7.4 and 60 degrees C. Then, the CO3Ap was mixed with a neutralized collagen gel and the CO3Ap-collagen mixtures with different kinds of CO3Ap contents and porosity were lyophilized into sponges. Scanning electron micrography (SEM) observation of CO3Ap-collagen sponges showed favorable pores for cell invasion. Approximately 50-300 microm size pores appeared to continue through the bulk. Higher magnification of the sponge showed a better adhesion between CO3Ap crystals and collagen. X-ray high-resolution microtomography revealed a clear image of the 3D structure of the sponges. The porosity of 0, 70 and 90%(w/w) CO3Ap-collagen sponges was 79.2 +/- 2.8%, 72.6 +/- 2.4% and 48.9 +/- 6.1%, respectively. The 70%(w/w) CO3Ap-collagen sponge appeared to be the most favorable biomaterial from the viewpoint of natural bone properties. Mouse osteoblast MC3T3-E1 cells were cultured in alphaMEM with 10% FCS for 2 weeks. Hematoxylin-eosin staining confirmed osteoblast cells invaded well into the CO3Ap collagen sponge. These sponges are expected to be used as hard tissue scaffold biomaterials for therapeutic uses. PMID- 14751744 TI - Multilayered peptide incorporated collagen tubules for peripheral nerve repair. AB - Successful nerve regeneration process was achieved with improved mechanical strength by crosslinking tubular nerve guides made up of collagen. The multilayered collagen sheets were prepared from laminar evaporation of collagen solution. Scanning electron micrograph of the collagen tubes crosslinked with glutaraldehyde (GTA), microwave irradiation showed porous, fibrillar structures of collagen filaments in these matrices. The mechanical property of the crosslinked collagen tubes was carried out by tensile strength measurements. Fourier transform infrared spectra of the collagen films show that the native triple helicity was unaltered during multilayered preparation. It was observed that the structural integrity is unaltered during the multilayer preparation. Microscopic analysis indicates that the tubule surface acts as a surface of adherence and proliferation for the sprouting axons from the cut proximal nerve stumps. Solute diffusion studies on these tubes indicate that they are highly porous to wide range of molecular sizes during regeneration. Among the two types of crosslinking, the microwave irradiated collagen conduits results in ample myelinated axons compared with GTA group, where we observed more unmyelinated axons. PMID- 14751745 TI - Novel biodegradable electrospun membrane: scaffold for tissue engineering. AB - Nonwoven fibrous matrixes have been widely used as scaffolds in tissue engineering, and modification of microstructure of these matrices is needed to organize cells in three-dimensional space with spatially balanced proliferation and differentiation required for functional tissue development. The objective of this study was fabrication of nanofibrous matrix from novel biodegradable poly(p dioxanone-co-L-lactide)-block-poly(ethylene glycol) (PPDO/PLLA-b-PEG) copolymer, and to examine cell proliferation, morphology of cell-matrix interaction with the electrospun nanofibrous matrix. The electrospun structure composed of PPDO/PLLA-b PEG fibers with an average diameters of 380 nm, median pore size 8 microm, porosity more than 80% and mechanical strength 1.4 MPa, is favorable for cell matrix interaction and supports the active biocompatibility of the structure. NIH 3T3 fibroblast cell seeded on the structure tend to maintain phenotypic shape and guided growth according to nanofiber orientation. Good capability of the nanofibrous structure for supporting the cell attachment and proliferation are observed. This novel biodegradable scaffold will be applicable for tissue engineering based upon its unique architecture, which acts to support and guide cell growth. PMID- 14751747 TI - Generation of porous microcellular 85/15 poly (DL-lactide-co-glycolide) foams for biomedical applications. AB - Porous 85/15 poly (DL-lactide-co-glycolide) or PLGA foams were produced by the pressure quench method using supercritical CO2 as the blowing agent. The rate of CO2 uptake and CO2 equilibrium concentration in PLGA at different processing conditions were studied by performing sorption experiments. The effects of saturation pressure and temperature on average cell size and relative density of the resulting foams were also studied. The time required to approach equilibrium exhibited a minimum with increasing saturation pressure. The diffusion coefficient and equilibrium concentration of CO2 in PLGA increased with an increasing pressure in an approximately linear relationship. Porous PLGA foams were generated with relative densities ranging from 0.107 to 0.232. Foams showed evidence of interconnected cells with porosities as high as 89%. The pore size ranged from 30 to 100 microm. PMID- 14751746 TI - The role of the CaCl2-guluronic acid interaction on alginate encapsulated betaTC3 cells. AB - Previously we demonstrated that alginate composition has a significant effect on the growth of encapsulated betaTC3 cells and consequently on the overall metabolic and secretory activities of the encapsulated cultures. Based on these results we postulated that the mechanical properties of alginate were not responsible for the observed effects but rather, changes in the strength of the alginate gel network caused by changes in the number of alginate strands held together in the "egg-box" model are responsible for the observed effects. In this study we address this hypothesis with a series of experiments in which the strength of this interaction is manipulated by varying the calcium concentration either at the time of gelation or during culture maintenance. Our data show that increasing the concentration of the CaCl2 solution used at the time of gelation, thus increasing the strength of the alginate gel network, impedes the growth characteristics of betaTC3 cells encapsulated in a high guluronic acid content alginate. This effect is amplified by maintaining a constant supply of calcium ions during culture thus sustaining the interaction between guluronic acid residues and calcium ions. However, preparations of betaTC3 cells encapsulated in an alginate with high mannuronic acid content are not affected by changes in CaCl2 concentration due to the low percentage of consecutive guluronic acid residues. Therefore, the present data show that the strength of the alginate gel network is an important factor that influences the growth characteristics of encapsulated cell preparations. PMID- 14751748 TI - Cholesteryl-grafted functional amphiphilic poly(N-isopropylacrylamide-co-N hydroxylmethylacrylamide): synthesis, temperature-sensitivity, self-assembly and encapsulation of a hydrophobic agent. AB - A thermally responsive amphiphilic grafted copolymer comprised of hydroxyl containing random poly(N-isopropylacrylamide-co-N-hydroxylmethylacrylamide) as the hydrophilic chain and cholesteryl groups as hydrophobic side arms was developed for the controlled release of hydrophobic drugs. The polymer was temperature-sensitive with a lower critical solution temperature of 40.5 degrees C and a critical micelle concentration of 4 mg/l. Dynamic light-scattering studies showed that the amphiphilic polymer self-assembled into micelles in aqueous media with their mean sizes in the range of 25-34 nm. Transmission electron microscope studies showed that the nanoparticles prepared from the micelle solutions exhibited multiple morphologies including unusual cubic and cuboids-like shapes, and normal spherical shapes, which could be controlled by the formation conditions. Wide-angle X-ray scattering studies showed that these nanoparticles were amorphous in nature but a small crystalline phase existed and the crystallinity of particles increased with the decrease of initial formation concentration. Pyrene was employed as a model hydrophobic agent to examine the encapsulation ability of the polymer with respect to hydrophobic agents in aqueous media. The loading level of the polymer with respect to pyrene was 4.4 mg/g, indicating that the thermally responsive amphiphilic polymer would be able to be used for the encapsulation of hydrophobic drugs. PMID- 14751749 TI - Calcium sulphate-based cements containing cephalexin. AB - Cephalexin containing gypsum and apatite/gypsum cements have been synthesised. The presence of cephalexin into the cements does not alter neither the physico chemical behaviour of the cements nor produce structural changes on them. These cements behave as drug delivery systems when soaked in simulated body fluid. The release of the drug is different depending on the composition. For gypsum cements, the cephalexin is quickly released, helped by a dissolution process of the matrix, whereas the drug release is more controlled by the hydroxyapatite presence in hydroxyapatite/gypsum samples. Apatite containing cements do not only show a different drug release process, also the paste viscosity is lower and a faster formation "in vitro" of an apatite-type layer on their surface is observed. PMID- 14751750 TI - Development of a calcium phosphate co-precipitate/poly(lactide-co-glycolide) DNA delivery system: release kinetics and cellular transfection studies. AB - One of the most common non-viral methods for the introduction of foreign deoxyribonucleic acid (DNA) into cultured cells is calcium phosphate co precipitate transfection. This technique involves the encapsulation of DNA within a calcium phosphate co-precipitate, particulate addition to in vitro cell culture, endocytosis of the co-precipitate, and exogenous DNA expression by the transfected cell. In this study, we fabricated a novel non-viral gene transfer system by adsorbing DNA, encapsulated in calcium phosphate (DNA/Ca-P) co precipitates, to biodegradable two- and three-dimensional poly(lactide-co glycolide) matrices (2D-DNA/Ca-P/PLAGA, 3D-DNA/Ca-P/PLAGA). Co-precipitate release studies demonstrated an initial burst release over the first 48 h. By day 7, approximately 96% of the initially adsorbed DNA/Ca-P co-precipitate had been released. This was followed by low levels of co-precipitate release for 42 days. Polymerase chain reaction was used to demonstrate the ability of the released DNA containing co-precipitates to transfect SaOS-2 cells cultured in vitro on the 3D DNA/Ca-P/PLAGA matrix and maintenance of the structural integrity of the exogenous DNA. In summary, a promising system for the incorporation and controlled delivery of exogenous genes encapsulated within a calcium phosphate co precipitate from biodegradable polymeric matrices has been developed and may have applicability to the delivery of therapeutic genes and the transfection of other cell types. PMID- 14751751 TI - Effects of preparative parameters on the properties of chitosan hydrogel beads containing Candida rugosa lipase. AB - The influences of the pH, tripolyphosphate (TPP) concentration, and ionic strength of the gelling medium on the entrapment efficiency, release, and activity of lipase in chitosan hydrogel beads were studied. A solution of Candida rugosa lipase was prepared in a 1.5% w/v chitosan and 1% (v/v) acetic acid medium, and dropped into a TPP solution. Release of lipase in pH 7.2 Tris buffer was monitored over 36 h using the micro BCA protein assay. The activity of the entrapped enzyme was assayed using the Sigma lipase activity method. Following preliminary studies, an experimental design was followed to develop mathematical models that describe bead characteristics as functions of the pH and the TPP concentration in the gelling medium. The pH and the TPP concentration each had an effect on the entrapment, retention, and activity of lipase. Entrapped lipase retained a high degree of activity in multiple reactions. The ionic strength, in the range studied, exerted a minimal effect on bead characteristics. Statistical analysis allowed optimization within the factor space with respect to maximizing the enzyme entrapment efficiency and activity, and also minimizing the amount released after 36 h in the Tris buffer. PMID- 14751752 TI - Mechanical behaviour of a new acrylic radiopaque iodine-containing bone cement. AB - In total hip replacement, fixation of a prosthesis is in most cases obtained by the application of methacrylic bone cements. Most of the commercially available bone cements contain barium sulphate or zirconium dioxide as radiopacifier. As is shown in the literature, the presence of these inorganic particles can be unfavourable in terms of mechanical and biological properties. Here, we describe a new type of bone cement, where X-ray contrast is obtained via the introduction of an iodine-containing methacrylate copolymer; a copolymer of methylmethacrylate and 2-[4-iodobenzoyl]-oxo-ethylmethacrylate (4-IEMA) is added to the powder component of the cement. The properties of the new I-containing bone cement (I cement) are compared to those of a commercially available bone cement, with barium sulphate as radiopacifier (B-cement). The composition of the I-cement is adjusted such that similar handling properties and radiopacity as for the commercial cement are obtained. In view of the mechanical properties, it can be stated that the intrinsic mechanical behaviour of the I-cement, as revealed from compression tests, is superior to that of B-cement. Concerning the fatigue behaviour it can be concluded that, though B-cement has a slightly higher fatigue crack propagation resistance than I-cement, the fatigue life of vacuum-mixed I cement is significantly better than that of B-cement. This is explained by the presence of BaSO4 clumps in the commercial cement; these act as crack initiation sites. The mechanical properties (especially fatigue resistance) of the new I cement warrant its further development toward clinical application. PMID- 14751753 TI - Intramedullary fixation of femoral cortical osteotomies with interlocked biodegradable self-reinforced poly-96L/4D-lactide (SR-PLA96) nails. AB - Femoral diaphyseal osteotomies of adult sheep were fixed with intramedullary (i.m.) biodegradable self-reinforced poly-96L/4D-lactide (SR-PLA96) nails (Phi 10.66-10.78 mmx195-199 mm) that were interlocked with four Phi 1.5 mm metallic Kirschner (K) wires. Hollow i.m. nails used in the pilot study resulted in implant failure and only two of these sheep were followed long term (18 months and 3 years). In the main study, solid nails were used and this resulted in consolidation in 5/6 cases and in one non-union due to failure of both proximal K wires at 3 weeks. Sheep in the main study were followed for 6 and 12 months. Femurs were studied radiographically, microradiographically, histologically and by oxytetracycline labelling. Good bone healing was achieved and histology showed only a mild tissue reaction to the implant at 6-18 months. At 3 years, the implant had almost degraded and numerous foamy macrophages were ingesting the disintegrated material. No accumulations of lymphocytes implying an immunological inflammatory reaction were seen. This investigation demonstrated that absorbable SR-PLA96 i.m. nails can be used together with interlocking metallic K wires in fixation of simple cortical osteotomies in large animals weighing up to 63 kg. The long-term biocompatibility should be studied in a larger population. PMID- 14751754 TI - Process development of an acellular dermal matrix (ADM) for biomedical applications. AB - The object of this study was to compare the extent of decellularization at each critical step of processing porcine skin to produce an acellular dermal matrix (ADM) for biomedical applications. The results demonstrated that the removal of epidermis using treatment with 0.25% trypsin for 18 h and 0.1% sodium dodecyl sulfate (SDS) for 12 h at room temperature was beneficial for the subsequent treatment to remove cells in the dermal structure. Lengthy incubation in 0.25% trypsin (12 h) and then 560 units/l Dispase (12 h) at 25 degrees C of small pieces of porcine skin from which the epidermis had been removed efficiently removed cells and cellular components from the skin. Histological examinations revealed that the epidermis, dermal fibroblasts, and epidermal appendages were completely removed by these treatments, and the basic dermal architecture of collagen bundles was that of a loose meshwork. Examinations by TEM showed that the characteristics of collagen fibers in the ADM were retained after complete removal of cells present under optimal conditions defined in this study. SDS-PAGE and size-exclusion HPLC revealed that collagen fibers in the ADM were mostly type I and showed two typical component peaks identified as oligomers and monomers, respectively. PMID- 14751755 TI - Enzymatic degradation of starch-based thermoplastic compounds used in protheses: identification of the degradation products in solution. AB - Apart from favourable physico-chemical and mechanical properties, the most important requirement for a biodegradable polymer to be used in medical applications is its biocompatibility and the non-cytotoxicity of its degradation products. Their combined effect should assure the safe material degradation under controlled kinetics. The present work analyses the degradation behaviour of blends of corn starch with poly(ethylene-vinyl alcohol) copolymer (SEVA-C). The characterization included long-term degradation trials on simulated physiological solution with alpha-amylase up to 200 days. The degradation solutions were analysed by several techniques. High-performance liquid chromatography (HPLC) and colorimetric methods were used to monitor the liberation of carbohydrate as a consequence of starch hydrolysis by alpha-amylase. The hydration degree was followed by thermogravimetric analysis (TGA). Several degradation products such as carbohydrates ranging from C6 to C18 were identified. After alpha-amylase action, biodegradation was more pronounced in the first 100 days, after which the biodegradation rate decreased probably due to the structure and porosity of the material. The action of alpha-amylase solely led to the starch degradation, in contrast with other assays without enzymes where no carbohydrates were found in the degradation solutions. PMID- 14751756 TI - Toll receptor families: structure and function. PMID- 14751757 TI - TLR signaling pathways. AB - Toll-like receptors (TLRs) have been established to play an essential role in the activation of innate immunity by recognizing specific patterns of microbial components. TLR signaling pathways arise from intracytoplasmic TIR domains, which are conserved among all TLRs. Recent accumulating evidence has demonstrated that TIR domain-containing adaptors, such as MyD88, TIRAP, and TRIF, modulate TLR signaling pathways. MyD88 is essential for the induction of inflammatory cytokines triggered by all TLRs. TIRAP is specifically involved in the MyD88 dependent pathway via TLR2 and TLR4, whereas TRIF is implicated in the TLR3- and TLR4-mediated MyD88-independent pathway. Thus, TIR domain-containing adaptors provide specificity of TLR signaling. PMID- 14751758 TI - Endotoxin recognition molecules, Toll-like receptor 4-MD-2. AB - Toll-like receptors (TLRs) are innate pathogen recognition molecules for microbial products. Lipopolysaccharide (LPS), a membrane constituent of Gram negative bacteria, is one of the most potent microbial products. LPS is recognized by TLR4 and MD-2. TLR4 is a transmembrane protein, the extracellular domain of which is composed of a protein motif called leucine-rich repeats (LRR). MD-2 is an extracellular molecule that is associated with the extracellular LRR of TLR4. MD-2 has a role in cell surface expression of TLR4 and interaction with LPS. TLR4-MD-2 contributes to containment of infections by Gram-negative bacteria by activating immune responses. PMID- 14751759 TI - Signal transduction pathways mediated by the interaction of CpG DNA with Toll like receptor 9. AB - Synthetic oligodeoxynucleotides (ODN) expressing non-methylated "CpG motifs" patterned after those present in bacterial DNA have characteristic immunomodulatory effects. CpG DNA is recognized as a pathogen-associated molecular pattern, and triggers a rapid innate immune response. CpG ODN are being harnessed for a variety of therapeutic uses, including as immune adjuvants, for cancer therapy, as anti-allergens, and as immunoprotective agents. The signal transduction pathway mediated by the engagement of CpG DNA with Toll-like receptor 9 (TLR9) is shared with other members of the TLR family. Recent studies demonstrate that formation and maturation of CpG DNA-containing endosomes are regulated by phosphatidylinositol 3 kinases and the Ras-associated GTP-binding protein, Rab5, which are essential for the initiation of TLR9-mediated signaling. PMID- 14751760 TI - Toll-like receptors and acquired immunity. AB - Toll-like receptors are a family of pattern recognition receptors (PRRs) that evolved to detect microbial infection. These receptors recognize conserved molecular products derived from different classes of microorganisms, including Gram-positive and -negative bacteria, fungi, protozoa and viruses. Following recognition of ligands TLRs initiate signaling events that result in acute innate responses. In addition, TLRs are responsible for initiation of adaptive immune responses against pathogen-derived antigens primarily through triggering dendritic cell activation. Control of adaptive immunity by TLRs is a complex phenomenon and much needs to be understood about how different TLRs tailor the outcome of adaptive immune responses to the advantage of the host. Although TLRs have evolved to induce protective immune responses, under some circumstances, activation of these receptors may lead to autoimmune diseases. PMID- 14751761 TI - Toll-like receptors and dendritic cells: for whom the bug tolls. AB - Recognition of molecular signatures of potential pathogens via toll-like receptors (TLRs) activates dendritic cells (DC), leading to the initiation of adaptive immunity. TLR signalling in DC causes an increase in display of MHC peptide ligands for T cell recognition, upregulation of co-stimulatory molecules important for T cell clonal expansion and secretion of immunomodulatory cytokines, which direct T cell differentiation into effectors. Remarkably, ligation of distinct TLRs can trigger differential cytokine production in a single DC type or result in different cytokines in distinct DC sub-types. Studying the complexity of DC responses to TLR ligands illuminates the link between innate recognition and adaptive immunity, paving the way for improved vaccines and strategies to induce tolerance to autoantigens or allografts. PMID- 14751762 TI - The role of Toll-like receptors in combating mycobacteria. AB - In the battle against infections with mycobacteria, the body employs components of both the innate and adaptive arms of the immune system. Toll-like receptors (TLRs) mediate the activation of cells of the innate immune system leading to dynamic functions including direct anti-microbial activity, induction of cytokine secretion, triggering dendritic cell maturation, and triggering apoptosis. Furthermore, TLR activation is capable of modulating the adaptive immune response with a bias towards a Th1 T-cell response. However, the activation of TLRs by mycobacteria may also provide a means of immune evasion. Therefore, the modulation of TLR activation can influence the ability to properly destroy invading pathogens such as mycobacteria. PMID- 14751763 TI - Sensing infection in Drosophila: Toll and beyond. AB - Drosophila has evolved a potent immune system that is somewhat adapted to the nature of infections through the selective activation of either one of two NF kappa B-like signalling pathways, the Toll and IMD (Immune deficiency) pathways. In contrast to the mammalian system, the Toll receptor does not act as a pattern recognition receptor (PRR) but as a cytokine receptor. The sensing of microbial infections is achieved by at least four PRRs that belong to two distinct families: the peptidoglycan recognition proteins (PGRPs) and the Gram-negative binding proteins (GNBPs)/beta-glucan recognition proteins (beta GRPs). PMID- 14751764 TI - A systems approach to dissecting immunity and inflammation. AB - The immune and inflammatory responses are extraordinarily complex, involving the dynamic interaction of a wide array of tissues, cells, and molecules. Traditional approaches are by and large reductionist, shying away from complexity, but providing detailed knowledge of circumscribed physiologic, cellular and molecular entities. The sequencing of the human genome, in concert with emerging genomic and proteomic technologies permits the definition of a complete and dynamic parts list of the immune and inflammatory systems. When harnessed with powerful new computational approaches, this will for the first time provide a comprehensive description of these complex biological processes. PMID- 14751765 TI - Hyperammonemia impairs long-term potentiation in hippocampus by altering the modulation of cGMP-degrading phosphodiesterase by protein kinase G. AB - Hyperammonemia impairs long-term potentiation (LTP) in hippocampus, by an unknown mechanism. LTP in hippocampal slices requires activation of the soluble guanylate cyclase (sGC)-protein kinase G (PKG)-cGMP-degrading phosphodiesterase pathway. The aim of this work was to assess whether hyperammonemia impairs LTP by impairing the tetanus-induced activation of this pathway. The tetanus induced a rapid cGMP rise, reaching a maximum at 10 s, both in the absence or presence of ammonia. The increase in cGMP is followed in control slices by a sustained decrease in cGMP due to PKG-mediated activation of cGMP-degrading phosphodiesterase, which is required for maintenance of LTP. Hyperammonemia prevents completely tetanus-induced cGMP decrease by impairing PKG-mediated activation of cGMP-degrading phosphodiesterase. Addition of 8Br-cGMP to slices treated with ammonia restores both phosphodiesterase activation and maintenance of LTP. Impairment of LTP in hyperammonemia may be involved in the impairment of the cognitive function in patients with hepatic encephalopathy. PMID- 14751766 TI - Microglial activation facilitates Abeta plaque removal following intracranial anti-Abeta antibody administration. AB - The mechanisms by which anti-Abeta antibodies clear amyloid plaques in Abeta depositing transgenic mice are unclear. In the current study, we demonstrate that inhibition of anti-Abeta antibody-induced microglial activation with anti inflammatory drugs, such as dexamethasone, inhibits removal of fibrillar amyloid deposits. We also show that anti-Abeta F(ab')(2) fragments fail to activate microglia and are less efficient in removing fibrillar amyloid than the corresponding complete IgG. Diffuse Abeta deposits are cleared by antibodies under all circumstances. These data suggest that microglial activation is necessary for efficient removal of compact amyloid deposits with immunotherapy. Inhibition of this activation may result in an impaired clinical response to vaccination against Abeta. PMID- 14751767 TI - Complement activation in human prion disease. AB - The central event in the neuropathological process of prion diseases (PrD) is the accumulation of abnormal prion protein accompanied by severe neuronal loss. Despite the infectious nature of these diseases, no prominent immune response has been detected yet. However, recent studies have shown that complement, a component of the innate immune system, is involved in the early pathogenesis of experimental prion infection. Here we demonstrate, in the diseased human brains, the presence of active compounds of the complement system, like C1q and C3b, in extracellular disease-associated prion protein deposits and the membrane attack complex in neurons. The neuronal localization of the membrane attack complex correlates well with the severity of disease-specific pathology and TUNEL labeling of neurons, irrespective of genotype or molecular phenotype of human prion diseases. PMID- 14751768 TI - Mitochondrial abnormalities in cybrid cell models of sporadic Alzheimer's disease worsen with passage in culture. AB - We created and studied new cybrid cell lines from sporadic Alzheimer's disease (SAD) or control (CTL) subjects to assess mitochondrial abnormalities just after metabolic selection ("early passage") and again six passages later ("late passage"). Cytochrome oxidase (CO) activities in early passage SAD cybrids created independently from the same platelet samples were highly correlated. Early passage SAD and CTL cybrids showed equivalent mitochondrial morphologies. Late passage SAD cybrids showed increased mitochondrial number, reduced mitochondrial size, and an approximately eightfold increase in morphologically abnormal mitochondria. Deficiency of SAD cybrid mitochondrial membrane potentials (DeltaPsi(M)) increased with passage. Mitochondrial bromodeoxyuridine (BrdU) uptake to estimate mitochondrial DNA (mtDNA) synthesis did not change with passage in CTL but increased in SAD cybrids. With time in culture, SAD mtDNA appears to replicate faster in cybrids, yielding cells with relative worsening of bioenergetic function. Metabolically deleterious SAD mitochondrial genes, like those in yeast, may have a replicative advantage over nondeleterious mitochondrial genes that assume dominance in CTL cybrids. PMID- 14751769 TI - Neuronal localization of C1q in preclinical Alzheimer's disease. AB - Complement has been postulated to contribute to inflammatory reactions associated with the neuropathology of Alzheimer's disease (AD). C1q, an initial component of the complement cascade, is associated with neuritic plaques and with neurons in the hippocampus of AD brain. Here, we report the presence of C1q in a cognitively intact subject, previously identified as preclinical AD. We compared in detail brain tissue of this preclinical case with a genetically related late-onset AD case. In the AD brain, C1q was typically associated with fibrillar Abeta plaques in frontal cortex and with plaques and neurons in the hippocampus. In the preclinical subject, C1q was abundantly present but it was cell-associated only, being primarily colocalized with neurons in both frontal cortex and hippocampus. However, no predominant cortical neuronal C1q localization was found in other preclinical cases or in Down's cases of different ages. Thus, it is possible that this neuronal-associated C1q reflects an early, but transient, response to injury that may modulate the progression of neurological dysfunction in AD. PMID- 14751770 TI - Characterisation of cytoskeletal abnormalities in mice transgenic for wild-type human tau and familial Alzheimer's disease mutants of APP and presenilin-1. AB - To study the role of Abeta amyloid deposits in the generation of cytoskeletal lesions, we have generated a transgenic mouse line coexpressing in the same neurons a wild-type human tau isoform (0N3R), a mutant form of APP (751SL) and a mutant form of PS1 (M146L). These mice developed early cerebral extracellular deposits of Abeta, starting at 2.5 months. A somatodendritic neuronal accumulation of transgenic tau protein was observed in tau only and in tau/PS1/APP transgenic mice, including in neurons adjacent to Abeta deposits. The phosphorylation status of this somatodendritic tau was similar in the two transgenic lines. The Abeta deposits were surrounded by a neuritic reaction composed of axonal dystrophic processes, immunoreactive for many phosphotau epitopes and for the human tau transgenic protein. Ultrastructural observation showed in these dystrophic neurites a disorganisation of the microtubule and the neurofilament network but animals that were observed up to 18 months of age did not develop neurofibrillary tangles. These results indicate that overexpression of mutant PS1, mutant APP and of wild-type human tau were not sufficient per se to drive the formation of neurofibrillary tangles in a transgenic model. The Abeta deposits, however, were associated to marked changes in cytoskeletal organisation and in tau phosphorylation in adjacent dystrophic neurites. PMID- 14751771 TI - Excitotoxic degeneration of hypothalamic orexin neurons in slice culture. AB - Several lines of evidence indicate that narcolepsy, a sleep disorder, results from the loss of hypothalamic orexin (hypocretin)-containing neurons, but the mechanisms responsible for selective elimination of this neuronal population are unknown. Using organotypic rat hypothalamic slice cultures, we investigated vulnerability of orexin neurons to excitotoxic insults. Twenty-four hours of incubation with N-methyl-D-aspartate (NMDA) followed by a recovery period of 72 h resulted in a marked decrease in the number of orexin-immunoreactive neurons, whereas melanin-concentrating hormone (MCH)-immunoreactive neurons in the same cultures were relatively spared. In contrast, orexin neurons were more resistant to kainic acid cytotoxicity than MCH neurons. Examinations of the effects of several endogenous glutamate receptor agonists as well as a glutamate transporter blocker highlighted quinolinic acid as an endogenous excitotoxin that could cause selective loss of orexin neurons as compared to MCH neurons by activating NMDA receptors. In addition, quinolinic acid-induced decrease of orexin neurons was prevented by an inhibitor of poly(ADP-ribose) polymerases. These results provide the first evidence concerning cytotoxic consequences onto orexin neurons, and indicate that NMDA receptor-mediated injury may contribute to the selective loss of these neurons in the hypothalamus, a prominent neuropathological feature found in narcolepsy patients. PMID- 14751772 TI - Spinal and cortical spreading depression enhance spinal cord activity. AB - Cortical spreading depression (CSD) has been suggested to underlie some neurological disorders such as migraine. Despite the intensity with which many investigators have studied SD in the brain, only a few studies have aimed to identify SD in the spinal cord. Here we described the main characteristic features of SD in the spinal cord induced by different methods including various spinal cord injury models and demonstrated that SD enhances the spinal cord activity following a transient suppressive period. These findings suggest that SD may play a role in the mechanisms of spinal neurogenic shock, spinal cord injury, and pain. Furthermore, we studied the effect of CSD on the neuronal activity of the spinal cord. CSD was induced via cortical pinprick injury or KCl injection in the somatosensory cortex. CSD did not propagate into the cervical spinal cord. However, intracellular recordings of the neurons in the dorsal horn of C2 segment, ipsilateral to the hemisphere in which CSD was evoked, showed a transient suppression of spontaneous burst discharges, followed by a significant enhancement of the neuronal activity. This indicates a link between a putative cause of the neurological symptoms and the subsequent pain of migraine. PMID- 14751773 TI - Biochemical and molecular studies of NMDA receptor subunits NR1/2A/2B in hippocampal subregions throughout progression of Alzheimer's disease pathology. AB - Alzheimer's disease (AD) is characterized by loss of specific cell populations within selective subregions of the hippocampus. Excitotoxicity, mediated via ionotropic glutamate receptors, may play a crucial role in this selective neuronal vulnerability. We investigated whether alterations in NMDA receptor subunits occurred during AD progression. Employing biochemical and in situ hybridization techniques in subjects with a broad range of AD pathology, protein levels, and mRNA expression of NR1/2A/2B subunits were assayed. With increasing AD neuropathology, protein levels and mRNA expression for NR1/2B subunits were significantly reduced, while the NR2A subunit mRNA expression and protein levels were unchanged. Cellular analysis of neuronal mRNA expression revealed a significant increase in the NR2A subunit in subjects with moderate neurofibrillary tangle neuropathology. This investigation supports the hypothesis that alterations occur in the expression of specific NMDA receptor subunits with increasing AD pathologic severity, which is hypothesized to contribute to the vulnerability of these neurons. PMID- 14751774 TI - Low brain glutathione and ascorbic acid associated with dopamine uptake inhibition during rat's development induce long-term cognitive deficit: relevance to schizophrenia. AB - Schizophrenia is associated with a cerebral glutathione deficit, which may leave the brain susceptible to oxidants. To study the consequences of a glutathione deficit, we treated developing rats with L-buthionine-(S,R)-sulfoximine (BSO), an inhibitor of glutathione synthesis, and later investigated their behaviour until adulthood. Since rodents may in some occasions compensate for a glutathione deficit by ascorbic acid (AA), we used Osteogenic Disorder Shionogi (ODS) mutant rats, which like humans, cannot synthetize ascorbic acid. Moreover, as hyperactivity of the dopaminergic system may be associated with schizophrenia, some rats were treated with the dopamine uptake inhibitor GBR 12909. Whereas ODS rats treated with either BSO or GBR 12909 alone had normal behaviour, rats treated with both BSO and GBR 12909 failed to discriminate between familiar and novel objects although other behaviours proved to be normal. In contrast, nonmutant rats were not affected by treatment with BSO and GBR 12909. Our results suggest that low brain glutathione and ascorbic acid levels associated with a perturbation of the dopaminergic system actively participate in the development of some cognitive deficits affecting schizophrenic patients. PMID- 14751775 TI - Suppression of neurite outgrowth by high-dose nerve growth factor is independent of functional p75NTR receptors. AB - We have previously demonstrated that high concentrations of nerve growth factor suppress neurite outgrowth from sensory neurons. Inhibition could be mediated by either the p75NTR or TrkA receptor. We used a functional block of p75NTR by REX antibody in rat dorsal root ganglion neurons and dorsal root ganglion cultures from p75NTR knockout mice. In both systems, high-dose NGF inhibited neurite outgrowth, implying that p75NTR is not involved in suppression of neurite outgrowth. Confocal images of dissociated dorsal root ganglion neurons exposed to fluorescence-tagged NGF showed ligand internalization. Radioligand binding indicated disappearance of high-affinity binding sites from the surface of dorsal root ganglia after treatment with 200 ng/ml NGF for 1 h. Downstream signaling showed sustained hyperphosphorylation of MAPK (Erk(1-2)) but not of SNT or Akt. High-dose NGF may induce cytoplasmic relocation of the receptor TrkA and axonal growth arrest independently of p75NTR. PMID- 14751776 TI - Positional effects of presenilin-1 mutations on tau phosphorylation in cortical plaques. AB - Mutations in presenilin-1 (PS-1) account for the majority of familial Alzheimer's disease (AD). While increasing Abeta42 is one mechanism whereby PS-1 mutations are thought to exert their pathogenic effect, little is known about the role of tau in PS-1 AD. This study compares staining (AT8 and tau-2), morphology and quantity of tau-immunoreactive cortical plaques in six PS-1 and five sporadic AD cases. The densities of tau-positive plaques differentiated PS-1 from sporadic AD cases. All PS-1 cases demonstrated a greater than 6-fold increase in tau-2 positive plaques. In PS-1 cases with mutations in exons 5 and 6, there was an increase in classical AD plaques containing hyperphosphorylated tau (AT8- and tau 2-positive). However, cases with exon 8 and 9 mutations had numerous cotton wool plaques containing nonhyperphosphorylated tau (tau-2-positive, AT8-negative). These findings suggest that PS-1 mutations increase tau deposition while mutation specific cellular responses determine phosphorylation events and may influence cell death mechanisms. PMID- 14751777 TI - Antiapoptotic signaling by a remyelination-promoting human antimyelin antibody. AB - Stabilizing the survival of oligodendrocytes and oligodendrocyte precursors within and near lesions in patients suffering from multiple sclerosis (MS) and other demyelinating diseases is an important therapeutic goal. Previous studies have identified a human-derived monoclonal IgM antibody designated rHIgM22 that induces remyelination in a mouse model of MS. We provide evidence that this antibody, directed against myelin, induces antiapoptotic signaling in premyelinating oligodendrocytes and reduces caspase-3 activation and caspase gene expression in mice undergoing antibody-induced remyelination. This effect was dependent on calcium entry via CNQX-sensitive channels and on lipid raft integrity, and was correlated with suppression of JNK signaling. We conclude that rHIgM22 may induce remyelination via rescue of oligodendrocytes, and suggest that such autoantibody-mediated signaling may have important therapeutic implications for a variety of neurological diseases, including stroke and Alzheimer's disease. PMID- 14751778 TI - Motor phenotypic alterations in TgDyrk1a transgenic mice implicate DYRK1A in Down syndrome motor dysfunction. AB - Motor deficits are among the most frequent impairments in Down syndrome (DS), but their neuropathological and molecular bases remain elusive. Here we investigate the motor profile of transgenic mice overexpressing Dyrk1a, Tg(Dyrk1a)1Cff (hereafter TgDyrk1a), a candidate gene hypothesized to cause some of the neurological defects associated with DS. We have previously shown DYRK1A expression in the cerebellum and functionally related structures, most brainstem motor nuclei and spinal cord, supporting a role for Dyrk1a in controlling motor function. Here we demonstrate that TgDyrk1a mice present DYRK1A overexpression in these areas along with specific motor dysfunction. The main finding that emerged was impairment of motor learning and alteration of the organization of locomotor behavior, which agrees with reported clinical observations in subjects with DS. These results confirm and extend previous data and provide further insight to the functional domains that might be altered in TgDyrk1a mice and underlying molecular mechanisms of DS motor dysfunction. PMID- 14751779 TI - Scrapie-infected GT1-1 cells show impaired function of voltage-gated N-type calcium channels (Ca(v) 2.2) which is ameliorated by quinacrine treatment. AB - Prions are transmissible pathogens that cause neurodegenerative diseases, although the mechanisms behind the nervous system dysfunctions are unclear. To study the effects of a prion infection on voltage-gated calcium channels, scrapie infected gonadotropin-releasing hormone neuronal cells (ScGT1-1) in culture were depolarized by KCl and calcium responses recorded. Lower calcium responses were observed in infected compared to uninfected cells. This effect was still observed when L-type calcium channels were blocked by nimodipine. After inhibition of N type calcium channels with omega-conotoxin GVIA, there was no difference in calcium responses. The calcium responses after nimodipine treatment became progressively lower during infection, but there was no major loss of the cellular prion protein (PrP(C)) or marked increase in accumulation of the abnormal prion protein (PrP(Sc)) in the cultures. These results indicate that scrapie infection causes a dysfunction of voltage-gated N-type calcium channels, which is exacerbated slowly over time. Quinacrine treatment cleared PrP(Sc) and restored calcium responses in the ScGT1-1 cultures. PMID- 14751780 TI - Death of cortical and striatal neurons induced by mitochondrial defect involves differential molecular mechanisms. AB - An important aspect of Huntington's disease (HD) pathogenesis which may have important therapeutic implications is that the cellular events leading to cell death may be different in cortical and striatal neurons. In the present study, we characterized cellular changes in cortical and striatal neurons treated with the mitochondrial toxin 3-nitropropionic acid (3NP) in culture. Degeneration induced by 3NP was similar in both striatal and cortical neurons as observed using markers of cell viability and DNA fragmentation. However, in striatal neurons, 3NP produced a marked delocalization of Bad, Bax, cytochrome c and Smac while this was not observed in cortical neurons. Death of striatal neurons was preceded by activation of calpain and was blocked by calpain inhibitor I. In cortical neurons, calpain was not activated and calpain inhibitor I was without effect. In both cell types, caspase-9 and -3 were not activated by 3NP and the caspase inhibitor zVAD-fmk did not provide neuroprotective effect. Interestingly, treatment with staurosporine (STS) triggered caspase-9 and -3 in cortical and striatal cells, suggesting that the molecular machinery related to caspase dependent apoptosis was functional in both cell types even though this machinery was not involved in 3NP toxicity. The present results clearly demonstrate that under mitochondrial inhibition, striatal and cortical neurons die through different pathways. This suggests that mitochondrial defects in HD may trigger the death of cortical and striatal neurons through different molecular events. PMID- 14751781 TI - Plasma and CSF markers of oxidative stress are increased in Parkinson's disease and influenced by antiparkinsonian medication. AB - We determined systemic oxidative stress in Parkinson's disease (PD) patients, patients with other neurological diseases (OND) and healthy controls by measurement of in vitro lipoprotein oxidation and levels of hydro- and lipophilic antioxidants in plasma and cerebrospinal fluid (CSF). Additionally, we investigated the influence of levodopa (LD) and dopamine agonist therapy (DA) on the oxidative status in PD patients. We found increased oxidative stress, seen as higher levels of lipoprotein oxidation in plasma and CSF, decrease of plasma levels of protein sulfhydryl (SH) groups and lower CSF levels of alpha-tocopherol in PD patients compared to OND patients and controls. Levodopa treatment did not significantly change the plasma lipoprotein oxidation but LD monotherapy tended to result in an increase of autooxidation and in a decrease of plasma antioxidants with significance for ubiquinol-10. DA monotherapy was significantly associated with higher alpha-tocopherol levels. Patients with DA monotherapy or co-medication with DA showed a trend to lower lipoprotein oxidation. These data support the concept of oxidative stress as a factor in the pathogenesis of PD and might be an indicator of a potential prooxidative role of LD and a possible antioxidative effect of DA in PD treatment. PMID- 14751782 TI - Restricted access materials and large particle supports for on-line sample preparation: an attractive approach for biological fluids analysis. AB - An analytical process generally involves four main steps: (1) sample preparation; (2) analytical separation; (3) detection; and (4) data handling. In the bioanalytical field, sample preparation is often considered as the time-limiting step. Indeed, the extraction techniques commonly used for biological matrices such as liquid-liquid extraction (LLE) and solid-phase extraction (SPE) are achieved in the off-line mode. In order to perform a high throughput analysis, efforts have been engaged in developing a faster sample purification process. Among different strategies, the introduction of special extraction sorbents, such as the restricted access media (RAM) and large particle supports (LPS), allowing the direct and repetitive injection of complex biological matrices, represents a very attractive approach. Integrated in a liquid chromatography (LC) system, these extraction supports lead to the automation, simplification and speeding up of the sample preparation process. In this paper, RAM and LPS are reviewed and particular attention is given to commercially available supports. Applications of these extraction supports, are presented in single column and column-switching configurations, for the direct analysis of compounds in various biological fluids. PMID- 14751783 TI - Validation study of assay method for DE-310, a novel polymer-bound camptothecin derivative, and the free drug in mouse plasma by liquid chromatography with fluorimetric detection. AB - DE-310 is a macromolecular carrier conjugate containing an anti-tumor camptothecin derivative, DX-8951, which is conjugated to a water-soluble polymer via a peptide spacer. Assay methods have been developed for the determination of a polymer-bonded DX-8951 conjugate, DX-8951, and Glycyl-DX-8951 (G-DX-8951) in mouse plasma. Free DX-8951 and Glycyl-DX-8951 were extracted from plasma by protein precipitation and analyzed by HPLC (Method I). Conjugated DX-8951 was extracted by protein precipitation and digested by using a thermolysin. The productive compound was analyzed by HPLC (Method II). The lower limits of quantitation of DX-8951, Glycyl-DX-8951, and Conjugated DX-8951 were 0.60, and 0.77 ng/ml and 3.45 microg/ml (as DX-8951 equivalent). These two methods showed satisfactory sensitivity, precision, accuracy, recovery, and selectivity. PMID- 14751784 TI - Simultaneous determination of 6beta-hydroxycortisol and cortisol in human urine by liquid chromatography with ultraviolet absorbance detection for phenotyping the CYP3A activity determined by the cortisol 6beta-hydroxylation clearance. AB - This study describes a high-performance liquid chromatographic (HPLC) method for the simultaneous determination of 6beta-hydroxycortisol (6beta-OHF) and cortisol in human urine using either methylprednisolone or beclomethasone as internal standard. Separation was achieved on a reversed-phase phenyl column by a gradient elution of 0.05 M KH(2)PO(4)-0.01 M CH(3)COOH (pH 3.77) and 0.05 M KH(2)PO(4) 0.01 M CH(3)COOH with acetonitrile (4:6, v/v). 6beta-Hydroxycortisol and cortisol were monitored by UV absorption at 239 nm. The lower quantitation limits of the present HPLC method were 21.5 ng/ml for 6beta-OHF and 5.0 ng/ml for cortisol in urine. The within-day reproducibilities in the amounts of 6beta-OHF and cortisol determined were in good agreement with the actual amounts added, the relative error being less than 1.59%. The inter-assay precisions (R.S.D. values) were less than 7.91% for 6beta-OHF and cortisol. The method was compared with the GC/MS method by measuring 6beta-OHF in the same urine samples. A good correlation was found between the amounts determined by the two methods. The regression equations for the HPLC (y) and GC/MS (x) methods were: y=1.0701x+17.389 (r=0.9772) for methylprednisolone as internal standard and y=1.0827x+6.1364 (r=0.9794) for beclomethasone as internal standard. PMID- 14751785 TI - Quantification and confirmation of flunixin in equine plasma by liquid chromatography-quadrupole time-of-flight tandem mass spectrometry. AB - The method describes quantification and confirmation of flunixin in equine plasma by liquid chromatography-quadrupole time-of-flight mass spectrometry (LC/Q TOF/MS/MS). Samples were screened by enzyme-linked immunosorbent assay (ELISA) and only those samples presumptively declared positive were subjected to quantification and confirmation for the presence of flunixin by this method. The method is also readily adaptable to instrumental screening for the analyte. Flunixin was recovered from plasma by liquid-liquid extraction (LLE). The sample was diluted with 2 ml saturated phosphate buffer (pH 3.10) prior to LLE. The dried extract was reconstituted in acetonitrile:water:formic acid (50:50:0.1, v/v/v) and subsequently analyzed on a Q-TOF tandem mass spectrometer (Micromass) operated under electrospray ionization positive ion mode. The concentration of flunixin was determined by the internal standard (IS) calibration method using the peak area ratio with clonixin as the IS. The limits of detection (LOD) and quantification (LOQ) for flunixin in equine plasma were 0.1 and 1 ng/ml, respectively, whereas the limit of confirmation (LOC) was 2.5 ng/ml. The qualifying ions for the identification of flunixin were m/z 297 [M+H](+), 279 (BP), 264, 259, 239 and those for clonixin (IS) were m/z 263 [M+H](+), 245 (BP) and 210. The measurement uncertainty about the result was 8.7%. The method is simple, sensitive, robust and reliably fast in the quantification and confirmation of flunixin in equine plasma. Application of this method will assist racing authorities in the enforcement of tolerance plasma concentration of flunixin in the racehorse on race day. PMID- 14751786 TI - Role of hydrophobicity on the monoamine receptor binding affinities of central nervous system drugs: a quantitative retention-activity relationships analysis using biopartitioning micellar chromatography. AB - Biological action and activity reflect an aspect of the fundamental physicochemical properties of the bioactive compounds. As an alternative to classical QSAR studies, in this work different quantitative retention-activity relationships (QRAR) models are proposed, which are able to describe the role of hydrophobicity on the binding affinity to different brain monoamine receptors (H(1)-histamine, alpha(1)-noradrenergic and 5-HT(2)-serotonergic) of different families of psychotherapeutic drugs. The retention of compounds is measured in a biopartitioning micellar chromatography (BMC) system using Brij-35 mobile phases. The adequacy of the QRAR models developed is due to the fact that both the retention of compounds in BMC and the drug-receptor interaction are described by the same hydrophobic, electronic and steric properties of compounds. The obtained results indicate that, for structurally related compounds that present the same molecular features as the basic pharmacophore, there is a retention range in which compounds present the highest affinity to all of monoamine receptors. PMID- 14751787 TI - Development and validation of a reversed-phase liquid chromatographic method for analysis of D4T (Stavudine) in rat plasma. AB - An improved and simplified high-performance liquid chromatographic (HPLC) method at UV detection 265 nm is presented for the determination of d4T in rat plasma. The mobile phase consists of methanol-distilled water-acetic acid in the 23:77:0.2 (v/v) ratio. Quantification is achieved by the peak-area ratio method with reference to the internal standard. This paper presents linearity, accuracy, precision, limit of quantification and limit of detection, specificity selectivity and sample stability data. Based on the intra and inter-day validation, all coefficients of variation (CV) were found less than 15%. The assay is sufficiently rapid and sensitive and was applied in a pharmacokinetic study in rats. PMID- 14751788 TI - Method for simultaneous measurement of norepinephrine, 3-methoxy-4 hydroxyphenylglycol and 3,4-dihydroxyphenylglycol by liquid chromatography with electrochemical detection: application in rat cerebral cortex and plasma after lithium chloride treatment. AB - An assay was developed to quantify norepinephrine (NE) and its metabolites (MHPG and DHPG) by high-performance liquid chromatography with electrochemical detection method (HPLC-ECD) in brain tissue and plasma of rats treated by LiCl. Separation on C(18) column was obtained by a mobile phase consisting of 4.5% methanol in buffer (0.1 M sodium acetate, 0.2 M citric acid) containing 0.2 mM ethylenediaminetetraacetic acid disodium salt (EDTA Na(2)) and 0.4 mM sodium octylsulfate, operated at a flow rate of 0.8 ml/min. A potential of +0.78 V was applied across the working and reference electrodes of the detector. The precision was in the range 2.88-4.35% for NE, 5.94-11.0% for MHPG and 1.97-4.40% for DHPG. Accuracy was 98.8-99.3% for NE, 97.4-100% for MHPG and 96.1-101% for DHPG. The limit of detection was 0.6 ng/ml for NE, 0.5 ng/ml for MHPG and 0.2 ng/ml for DHPG. The linearity is over the range 20-60 ng/ml for NE, 7-23 ng/ml for MHPG and 6-20 ng/ml for DHPG. The assay has been applied successfully to measure simultaneously cortex and plasmas concentrations of these three catecholamines in rats. PMID- 14751789 TI - Urine sample preparation of tricyclic antidepressants by means of a supported liquid membrane technique for high-performance liquid chromatographic analysis. AB - Supported liquid membrane (SLM) technique for sample work-up and enrichment was used for determination of tricyclic antidepressant drugs in urine by high performance liquid chromatography (HPLC) with UV detection. The studied antidepressant drugs were amitriptyline, opipramol, noxiptyline and additionally diethazine was used as possible internal standard. Alkaline phosphoric buffer with urine sample, as the donor solution, was passed over the liquid membrane into which investigated substances were extracted. On the other side of the membrane, analyzed compounds were trapped due to creating non-extractable form in acidic acceptor solution. Enriched and cleaned up drugs were then injected into a HPLC system with ultraviolet detection to analyze of their concentration in acceptor solution. Optimum extraction efficiency was determined by changing acceptor and donor solutions pH, application of different flow rates of donor solution and by using different solvents in the membrane. Also, donor solution volume, extraction time and concentration of analytes were varied to check the linearity of extraction process. The highest extraction efficiency: 43% for opipramol, 56% for noxiptyline, 43% for amitriptyline and 42% for diethazine (R.S.D. values were <6% and n=3) was achieved when 0.05 M phosphate buffer pH 4.0 and 9.5 were used as donor and acceptor solutions, respectively, n-undecane with 5% tri-n-octylphosphine oxide (TOPO) was used as liquid membrane. Limit of quantification (LOQ) for tricyclic antidepressants after enrichment of 100ml of urine sample was about 1 ng/ml. PMID- 14751790 TI - Anti-coagulant rodenticide binding properties of human serum albumin: a biochromatographic approach. AB - In this paper, the anti-coagulant rodenticide-human serum albumin (HSA) binding was investigated using a perturbation method to calculate the solute distribution isotherms. It was shown that rodenticide can bound either on the benzodiazepine HSA site with low affinity (site I) or on the warfarin HSA site with high affinity (site II). The thermodynamic parameters of this association were calculated for the two HSA binding sites. For the site II, the rodenticide-HSA association was governed enthalpically whereas for the site I, this one was driven entropically. Moreover, the role of the magnesium (Mg(2+)) and calcium (Ca(2+)) on this association was carried out. It was clearly demonstrated that the rodenticide affinity for the site I was not affected by modifying the bulk solvent surface tension whereas for the site II the association constant increased strongly with the Mg(2+) or the Ca(2+) concentration in the bulk solvent. These results showed that the rodenticide-HSA affinity and thus the rodenticide toxicological effect depends on the Mg(2+) or Ca(2+) concentration. PMID- 14751791 TI - Comparison of measured and calculated lipophilicity of substituted aurones and related compounds. AB - A molecule library containing 55 aurone- and thioaurone-type structures has been designed and synthesised. Reversed phase high performance liquid chromatographic (RP-HPLC) method has been developed to separate these compounds and to characterise their lipophilicity by experimental method (k'). The experimental lipophilicity data have been compared with the computer calculated lipophilicity parameters (CLOGPs) of the same molecules. In general, good correlations between the measured and calculated lipophilicities have been found with the exception of structure isomers and compounds capable for hydrogen bonding. The chromatographic method was suitable to separate the structure (ortho and para) isomers of aurone and thioaurones and was sensitive enough to differentiate their lipophilicities. Our findings suggest the usefulness of the chromatographic method in fast characterisation of the lipophilicity of structurally closely related molecules. PMID- 14751792 TI - Determination of queuosine derivatives by reverse-phase liquid chromatography for the hypomodification study of Q-bearing tRNAs from various mammal liver cells. AB - Three queuosine derivatives (Q-derivatives) have been found at position 34 of four mammalian so-called Q-tRNAs: queuosine (Q) in tRNA(Asn) and tRNA(His), mannosyl-queuosine (manQ) in tRNA(Asp), and galactosyl-queuosine (galQ) in tRNA(Tyr). An analytical procedure based on the combined means of purified tRNA isolation from liver cells and ribonucleoside analysis by reverse-phase high performance liquid chromatography coupled with real-time UV-spectrometry (RPLC UV) was developed for the quantitative analysis of the three Q-derivatives present in total tRNA from liver tissues and liver cell cultures. Using this analytical procedure, the rates of Q-tRNA modification were studied in total tRNAs from various mammalian hepatic cells. Our results show that the four Q tRNAs are fully modified in liver tissues from adult mammals, regardless of the mammal species. However, a lack in the Q-modification level was observed in Q tRNAs from newborn rat liver, as well in Q-tRNAs from normal rat liver cell cultures growing in a low queuine content medium, and from a rat hepatoma cell line. It is noteworthy that in all cases of Q-tRNA hypomodification, our analytical procedure showed that tRNA(Asp) is always the least affected by the hypomodification. The biological significance of this phenomenon is discussed. PMID- 14751793 TI - Galactonate determination in urine by stable isotope dilution gas chromatography mass spectrometry. AB - A stable isotope dilution assay was developed for the sensitive determination of D-galactonic acid. D-[U-13C(6)]galactono-1,4-lactone was prepared as internal standard. Unlabelled and U-13C-labelled D-galactonic acid species were converted to the N-(1-butyl)galactonamide pentaacetate derivatives and assessed by gas chromatography-mass spectrometry (GC-MS). Positive chemical ionisation and monitoring of the [MH-60](+)-ions in the galactonate chromatographic peak at m/z 402 and m/z 408 were used for quantification. The procedure was applied to study the variability of D-galactonate excretion in healthy subjects and galactosemic patients and to monitor the D-galactonate-D-galactitol ratio in human urine. PMID- 14751794 TI - Simple and sensitive liquid chromatography-tandem mass spectrometry method for determination of the S(+)- and R(-)-enantiomers of baclofen in human plasma and cerebrospinal fluid. AB - A simple and sensitive liquid chromatography-tandem mass spectrometry (LC/MS/MS) method to determine the enantiomers of the muscle relaxant baclofen in human plasma and cerebrospinal fluid (CSF) has been developed. A commercially available ultrafiltration membrane is used to prepare the sample. A chiral CROWNPAK CR(+) stationary phase column is then used to perform complete resolution of the S(+)- and R(-)-enantiomers of baclofen. This method was used to analyze human plasma and CSF spiked with baclofen, and the calibration curves for both biologic samples were linear over a concentration range of 0.15-150 ng enantiomer/ml. The lower limit of quantification was 0.15 ng enantiomer/ml in both fluids. Finally, the method was tested with an artificial CSF as an alternative to authentic human CSF. The results showed that no matrix effects and no interfering peaks were observed using this artificial CSF. PMID- 14751795 TI - Simultaneous determination of glipizide and rosiglitazone unbound drug concentrations in plasma by equilibrium dialysis and liquid chromatography-tandem mass spectrometry. AB - Glipizide and rosiglitazone are widely used to treat Type 2 diabetes. In order to investigate drug-drug protein binding interaction between glipizide and rosiglitazone, a method was developed and validated for simultaneously determining the free (unbound) fraction of glipizide and rosiglitazone in plasma employing equilibrium dialysis for the separation of free drug and liquid chromatography-tandem mass spectrometry (LC-MS/MS) for quantitation. Post dialysis human plasma or buffer samples of 0.2 ml were extracted using a liquid liquid extraction procedure and analyzed by a high performance liquid chromatography electrospray tandem mass spectrometer system. The compounds were eluted isocratically on a Zorbax SB-Phenyl column, ionized using an atmospheric pressure electrospray ionization source and analyzed in positive ion mode with multiple reaction monitoring. The ion transitions monitored were m/z 446-->321 for glipizide, m/z 358-->135 for rosiglitazone, and m/z 271-->155 for tolbutamide (internal standard, IS). The chromatographic run time was 5 min per injection, with retention times of 2.3, 3.4 and 2.3 min for glipizide, rosiglitazone and IS, respectively. The calibration curves of glipizide and rosiglitazone were over the range of 1-2000 ng/ml (r(2)>0.9969) in the combined matrix of human plasma and isotonic sodium phosphate buffer (1:1, v/v). The inter-assay precision and accuracy of the quality control samples were <10.9% of coefficient of variability and >93.5% and 94.5% of nominal concentration for glipizide and rosiglitazone, respectively. The lower limit of quantitation of both glipizide and rosiglitazone was 1.0 ng/ml. Both glipizide and rosiglitazone bound to plasma protein extensively (>99% bound). Glipizide and rosiglitazone free fraction averaged 0.678+/-0.071 and 0.389+/-0.061%, respectively, at plasma concentration of 1000 ng/ml. This developed method proves reproducible and sensitive and its application to clinical samples is also reported. PMID- 14751796 TI - Confirmatory analysis of 17beta-boldenone, 17alpha-boldenone and androsta-1,4 diene-3,17-dione in bovine urine, faeces, feed and skin swab samples by liquid chromatography-electrospray ionisation tandem mass spectrometry. AB - The origin, i.e. natural occurrence or illegal treatment, of findings of 17alpha boldenone (alpha-Bol) and 17beta-boldenone (beta-Bol) in urine and faeces of cattle is under debate within the European Union. A liquid chromatographic positive ion electrospray tandem mass spectrometric method is presented for the confirmatory analysis of 17beta-boldenone, 17alpha-boldenone and an important metabolite/precursor androsta-1,4-diene-3,17-dione (ADD), using deuterium labelled 17beta-boldenone (beta-Bol-d3) as internal standard. Detailed sample preparation procedures were developed for a variety of sample matrices such as bovine urine, faeces, feed and skin swab samples. The method was validated as a quantitative confirmatory method according to the latest EU guidelines and shows good precision, linearity and accuracy data, and CCalpha and CCbeta values of 0.1 0.3 and 0.4-1.0 ng/ml, respectively. Currently, the method has been successfully applied to suspect urine samples for more than a year, and occasionally to faeces, feed and swab samples as well. Results obtained from untreated and treated animals are given and their impact on the debate about the origin of residues of 17beta-boldenone is critically discussed. Finally, preliminary data about the degree of conjugation of boldenone residues are presented and a simple procedure for discrimination between residues from abuse versus natural origin is proposed. PMID- 14751797 TI - Determination of Astragaloside IV in rat plasma by liquid chromatography electrospray ionization mass spectrometry. AB - Astragaloside IV (AGS-IV) is an active constituent of Radix Astragali used in many Traditional Chinese Medicines. This paper describes a sensitive and specific assay for the quantitation of AGS-IV in rat plasma. After solid phase extraction (SPE), samples were analyzed by liquid chromatography electrospray ionization mass spectrometry using a reversed-phase C18 column. The assay was linear in the range 1-500 ng/ml with a limit of detection of 0.5 ng/ml. The recovery was 92.5% and within-day and between-day precision were 3.7-6.0 and 2.8-9.8%, respectively. The assay was applied to a pharmacokinetic study in rat after a single oral dose. The drug was rapidly absorbed and subsequently eliminated according to a biphasic concentration-time curve. PMID- 14751798 TI - Rapid screening procedure based on headspace solid-phase microextraction and gas chromatography-mass spectrometry for the detection of many recreational drugs in hair. AB - An increasing number of synthetic drugs are appearing on the illicit market and on the scene of drug use by youngsters. Official figures are underestimated. In addition, immunochemical tests are blind to many of these drugs and appropriate analytical procedures for routine clinical and epidemiological purposes are lacking. Therefore, the perceived increasing abuse of recreational drugs has not been proved yet. In a previous paper, we proposed a procedure for the preliminary screening of several recreational substances in hair and other biological matrices. Unfortunately, this procedure cannot apply to cocaine. Consequently, we performed a new headspace solid-phase microextraction and gas chromatography-mass spectrometry (HS-SPME-GC-MS) procedure for the simultaneous detection of cocaine, amphetamine (A), methamphetamine (MA), methylen-dioxyamphetamine (MDA), methylen dioxymethamphetamine (MDMA), methylen-dioxyethamphetamine (MDE), N-methyl-1-(1,3 benzodioxol-5-yl)-2-butanamine (MBDB), ketamine, and methadone in human hair. Hair was washed with water and acetone in an ultrasonic bath. A short acid extraction with 1M hydrochloric acid was needed; the fiber was exposed to a 5 min absorption at 90 degrees C and thermal desorption was performed at 250 degrees C for 3 min. The procedure was simple, rapid, required small quantities of sample and no derivatization. Good linearity was obtained over the 0.1-20.0 ng/mg range for the target compounds. Sensitivity was good enough: limits of detection (LOD) were 0.7 ng/mg of hair for the majority of substances. The intra-day precision ranged between 7 and 20%. This paper deals with the analytical performance of this procedure and its preliminary application to hair samples obtained on a voluntary basis from 183 young people (138 males and 45 females) in the Rome area. PMID- 14751799 TI - Multidimensional on-line sample preparation of verapamil and its metabolites by a molecularly imprinted polymer coupled to liquid chromatography-mass spectrometry. AB - A new molecularly imprinted polymer (MIP) material was synthesized selective for verapamil and utilized for on-line metabolic screening of this common calcium antagonist in biological samples. Since some metabolites of verapamil have also shown pharmacological properties, a selective and sensitive sample preparation approach that provides a metabolic profile in biologically relevant samples is important. The MIP material was coupled on-line to a restricted access material (RAM) precolumn. The multidimensional nature of this set-up removed large matrix interferents such as proteins from the sample, while the selectivity of the MIP enabled further cleanup of the smaller analytes. The selectivity and extraction efficiency of the MIP for verapamil and its metabolites was evaluated in various biological matrices, such as cell cultures and urine. The experimental set-up with the developed method enabled the direct injection of biological samples for the selective isolation, preconcentration, identification and analysis of verapamil and its phase I metabolites by LC-MS(n). This multidimensional approach provided much qualitative information about the metabolic profile of verapamil in various biological matrices. An analytical method was developed for the quantification of verapamil and gallopamil in urine, plasma and cell culture. Acceptable linearity (R(2)=0.9996, 0.9982 and 0.9762) with an average injection repeatability (n=3) of 10, 25 and 15% R.S.D. was determined for urine, plasma and cell culture, respectively. This is the first application of the procedure for the selective metabolic screening of verapamil in biological samples. PMID- 14751800 TI - Simultaneous determination of 1- and 2-naphthol in human urine using on-line clean-up column-switching liquid chromatography-fluorescence detection. AB - We developed a new 3-D HPLC method for on-line clean-up and simultaneous quantification of two important naphthalene metabolites, 1-naphthol and 2 naphthol, in human urine. Except an enzymatic hydrolysis no further sample pre treatment is necessary. The metabolites are stripped from urinary matrix by on line extraction on a restricted access material pre-column (RAM RP-8), transferred in backflush mode onto a silica-based CN-(cyano)phase column for further purification from interfering substances. By another successive column switching step both analytes are transferred with a minimum of overlapping interferences onto a C12 bonded reversed phase column with trimethylsilyl endcapping where the final separation is carried out. The entire arrangement is software controlled. Eluting analytes are quantified by fluorescence detection (227/430 nm) after an external calibration. Within a total run time of 40 min we can selectively quantify both naphthols with detection limits in the lower ppb range (1.5 and 0.5 microg/l for 1- and 2-naphthol, respectively) with excellent reliability (ensured by precision, accuracy, matrix-independency and FIOH quality assurance program participation). First results on a collective of 53 occupationally non exposed subjects showed mean levels of 11.0 microg/l (1 naphthol) and 12.9 microg/l (2-naphthol). Among smokers (n=21) a significantly elevated mean level of urinary naphthols was determined (1-naphthol: 19.2 microg/l and 2-naphthol: 23.7 microg/l) in comparison to non smokers (n=32; 1 naphthol: 5.6 microg/l, 2-naphthol: 5.6 microg/l). PMID- 14751801 TI - New trend in sample preparation: on-line microextraction in packed syringe for liquid and gas chromatography applications. I. Determination of local anaesthetics in human plasma samples using gas chromatography-mass spectrometry. AB - A new technique for sample preparation on-line with LC and GC-MS assays was developed. Microextraction in a packed syringe (MEPS) is a new miniaturised, solid-phase extraction technique that can be connected on-line to GC or LC without any modifications. In MEPS approximately 1mg of the solid packing material is inserted into a syringe (100-250 microl) as a plug. Sample preparation takes place on the packed bed. The bed can be coated to provide selective and suitable sampling conditions. The new method is very promising. It is very easy to use, fully automated, of low cost and rapid in comparison with previously used methods. This paper presents the development and validation of a method for microextraction in packed syringe MEPS on-line with GC-MS. Local anaesthetics in plasma samples were used as model substances. The method was validated and the standard curves were evaluated by the means of quadratic regression and weighted by inverse of the concentration: 1/x for the calibration range 5-2000 nM. The applied polymer could be used more than 100 times before the syringe was discarded. The extraction recovery was between 60 and 90%. The results showed close correlation coefficients (R>0.99) for all analytes in the calibration range studied. The accuracy of MEPS-GC-MS was between 99 and 115% and the inter-day precision (n=3 days), expressed as the relative standard deviation (R.S.D.%), was 3-10%. PMID- 14751802 TI - Analysis of catecholamines and related substances using porous graphitic carbon as separation media in liquid chromatography-tandem mass spectrometry. AB - Capillary porous graphitic carbon (PGC) columns have been utilized for separation of several catecholamines and related compounds (i.e. L-tyrosine, L-DOPA, 3-O methyl-DOPA, dopamine, 3,4-dihydroxy-phenyl-acetic acid (DOPAC), homovanillic acid, noradrenaline, vanillomandelic acid and adrenaline) on-line with electrospray ionization tandem mass spectrometry (ESI-MS/MS). The use of a mobile phase without ion-pairing agents and with high content of organic modifier facilitated the coupling to the selective and sensitive mass spectrometric detection. Minimum detectable sample concentration (MDC sample) for noradrenaline, dopamine and L-tyrosine in a standard solution was estimated to 3, 10 and 30 nM, respectively (3 S/N corresponds to MDQ for L-tyrosine of approximately 8 x 10(-14)mol). The developed strategy was applied for analysis of brain tissue, i.e. a substantia nigra (ns) sample. PMID- 14751803 TI - Application of short monolithic columns for fast purification of plasmid DNA. AB - Anion-exchange chromatography is one of the most important methods in downstream processing of plasmid DNA, both as a process and as an analytical technique. Separation of plasmid DNA on traditional particle-based anion-exchange supports is usually slow. Moreover, such supports have a low capacity for plasmid DNA due to the steric exclusion effects. In this work, the separation of plasmid DNA using short monolithic columns, Convective Interaction Media, will be presented. It will be demonstrated that plasmid DNA can be purified from bacterial cells using alkaline lysis followed by chromatography on a very short weak anion exchange chromatographic columns-disks-with good purity and quality within a short time. Furthermore, the separation of plasmid DNA from cell RNA can be carried out without the need of adding RNAse. Fast and efficient method for in process control of the purified plasmid will be described as well. PMID- 14751804 TI - Simultaneous determination of the acid/base antihypertensive drugs celiprolol, bisoprolol and irbesartan in human plasma by liquid chromatography. AB - A simple, rapid method for the simultaneous determination of cardiovascular drugs: celiprolol, bisoprolol and irbesartan in human plasma is described. The two main features of the proposed method deal first, with a simultaneous solid phase extraction of weakly basic beta-blockers derivatives and irbesartan which exhibit weak acidic properties; second with an absorbance monitoring using diode array detection in order to insure an improved selectivity. The separation is performed on a C(18) Kromasil 4.6 mm x 150 mm column using a linear gradient to achieve an entire separation of the four species in less than 20 min. The full analytical validation is performed according to guidance for industry for bioanalytical method validation. Linearity of the response was demonstrated for each drug for a range fulfilling the reported plasma levels, that is 10-500, 5 250 and 20-1000 ng l(-1) for celiprolol, bisoprolol and irbesartan respectively. Intra- and inter-day relative standard deviations for all compounds were, in any case, lower than 11% and the method exhibits a convenient accuracy (percentage of relative error lower than 6% for each drug). In each case, the LOD were sufficient to detect post dose trough concentrations for checking patient's observance. Moreover, selectivity towards either endogenous species or co administered drugs was demonstrated by combination of the use of the solid phase extraction process, gradient elution and diode array detection facilities, making thus, the proposed technique especially suitable for routine drug monitoring of resistant hypertensive patients. PMID- 14751805 TI - Determination of corticosterone and 17-hydroxycorticosterone in plasma and urine samples by sweeping techniques using micellar electrokinetic chromatography. AB - The analysis of corticosterone in mouse blood serum (metabolic-stress experiment) and 17-hydroxycorticosterone in human urine (exercise-stress experiment) samples by means of capillary electrophoresis/UV absorbance in conjunction with online sample concentration techniques is described. The use of normal MEKC had an analyte detection limit of 7 microg/ml (S/N=3); whereas when online sample concentration methods, including sweeping-micellar electrokinetic chromatography (Sweeping-MEKC) and cation-selective exhaustive injection-sweep-micellar electrokinetic chromatography (CSEI-sweep-MEKC) were used, the detection limits could be improved to 3 and 5 ng/ml, respectively. In the analysis of actual samples from animal metabolic-stress experiments (39 mouse), chronically stressed animals showed a higher level (552+/-152 ng/ml) and acute stressed animals showed an intermediate level (375+/-105 ng/ml). In comparison, normal animals show a lower concentration level of corticosterone (153+/-109 ng/ml). In addition, based on a human exercise-stress experiment (seven volunteers), the acute stressed humans (after exercise, 800 m of running) show a higher concentration of 17 hydroxycorticosterone (113+/-55 ng/ml for males; 128+/-25 for females) and the non-stressed humans (before exercise) show a lower concentration (63+/-37 ng/ml for male; 60+/-20 for female), respectively. PMID- 14751806 TI - Quantitative analysis of human salivary glucose by gas chromatography-mass spectrometry. AB - A reference analytic methodology was developed for the determination of human salivary glucose concentration. The technique involves the glucose derivatization with acetic anhydride and subsequent analysis of glucose penta-acetylated by gas chromatography combined with mass spectrometry. Glucose concentration in the biological fluid depends on the physiological status of the donor. PMID- 14751807 TI - Determination of 3-amino-5-mercapto-1,2,4-triazole in serum. AB - A high performance liquid chromatography (HPLC) method using fluorescence detection to determine 3-amino-5-mercapto-1,2,4-triazole (AMT) levels in serum has been developed. Sample preparation involved treatment with tributylphosphine (TBP) to reduce disulfides formed during storage, precipitation of proteins with acetonitrile (ACN), and precolumn derivatization using the thiol reactive fluorescent probe monobromobimane (MBB). The conjugate (AMT-MBB) was resolved by gradient elution from a C(18) reversed-phase column. The assay method was linear over a concentration range of 0.78-50 microg/ml and had a limit of detection (LOD) of 0.05 microg/ml AMT (10 microl injection). This method provides a sensitive and specific tool for the determination of AMT in serum and may have potential industrial hygiene application. PMID- 14751808 TI - Validated high-performance liquid chromatographic method utilizing solid-phase extraction for the simultaneous determination of naringenin and hesperetin in human plasma. AB - Naringenin and hesperetin, the aglycones of the flavanone glucosides naringin and hesperidin occur naturally in citrus fruits. They exert a variety of pharmacological effects such as antioxidant, blood lipid-lowering, anticarcinogenic and inhibit selected cytochrome P-450 enzymes resulting in drug interactions. A specific, sensitive, precise, and accurate solid-phase extraction high-performance liquid chromatographic (HPLC) assay for the simultaneous determination of naringenin and hesperetin in human plasma was developed and validated. After addition of 7-ethoxycoumarin as internal standard, plasma samples were incubated with beta-glucuronidase/sulphatase, and the analytes were isolated from plasma by solid-phase extraction using C(18) cartridges and separated on a C(8) reversed phase column with methanol/water/acetic acid (40:58:2, v/v/v) as the eluent at 45 degrees C. The method was linear in the 10 300 ng/ml concentration range for both naringenin and hesperetin (r>0.999). Recovery for naringenin, hesperetin and internal standard was greater than 76.7%. Intra- and inter-day precision for naringenin ranged from 1.4 to 4.2% and from 1.9 to 5.2%, respectively, and for hesperetin ranged from 1.3 to 4.1% and from 1.7 to 5.1%, respectively. Accuracy was better than 91.5 and 91.3% for naringenin and hesperetin, respectively. PMID- 14751809 TI - Peroxisome proliferator-activated receptor alpha agonists increase nitric oxide synthase expression in vascular endothelial cells. AB - OBJECTIVE: There has been accumulating evidence demonstrating that activators for peroxisome proliferator-activated receptor alpha (PPARalpha) have antiinflammatory, antiatherogenic, and vasodilatory effects. We hypothesized that PPARalpha activators can modulate endothelial nitric oxide synthase (eNOS) expression and its activity in cultured vascular endothelial cells. METHODS AND RESULTS: Bovine aortic endothelial cells were treated with the PPARalpha activator fenofibrate. The amount of eNOS activity and the expression of eNOS protein and its mRNA were determined. Our data show that treatment with fenofibrate for 48 hours resulted in an increase in eNOS activity. Fenofibrate failed to increase eNOS activity within 1 hour. Fenofibrate also increased eNOS protein as well as its mRNA levels. RU486, which has been shown to antagonize PPARalpha action, inhibited the fenofibrate-induced upregulation of eNOS protein expression. WY14643 and bezafibrate also increased eNOS protein levels, whereas rosiglitazone did not. Transient transfection experiments using human eNOS promoter construct showed that fenofibrate failed to enhance eNOS promoter activity. Actinomycin D studies demonstrated that the half-life of eNOS mRNA increased with fenofibrate treatment. CONCLUSIONS: PPARalpha activators upregulate eNOS expression, mainly through mechanisms of stabilizing eNOS mRNA. This is a new observation to explain one of the mechanisms of PPARalpha-mediated cardiovascular protection. PMID- 14751810 TI - Absence of dystrophin in mice reduces NO-dependent vascular function and vascular density: total recovery after a treatment with the aminoglycoside gentamicin. AB - OBJECTIVE: Mutations in the dystrophin gene causing Duchenne's muscular dystrophy (DMD) lead to premature stop codons. In mice lacking dystrophin (mdx mice), a model for DMD, these mutations can be suppressed by aminoglycosides such as gentamicin. Dystrophin plays a role in flow (shear stress)-mediated endothelium dependent dilation (FMD) in arteries. We investigated the effect of gentamicin on vascular contractile and dilatory functions, vascular structure, and density in mdx mice. METHODS AND RESULTS: Isolated mice carotid and mesenteric resistance arteries were mounted in arteriographs allowing continuous diameter measurements. Mdx mice showed lower nitric oxide (NO)-dependent FMD and endothelial NO synthase (eNOS) expression as well as decreased vascular density in gracilis and cardiac muscles compared with control mice. Treatment with gentamycin restored these parameters. In contrast, smooth muscle-dependent contractions as well as endothelium-dependent or -independent dilation were not affected by dystrophin deficiency or by gentamicin treatment. CONCLUSIONS: Dystrophin deficiency induces a selective defect in flow-dependent mechanotransduction, thus attenuating FMD and eNOS expression, and may contribute to low arteriolar density. These findings open important perspectives regarding the mechanism involved in the pathophysiology of genetic diseases related to premature stop codons such as DMD. PMID- 14751811 TI - Human apolipoprotein A-IV reduces secretion of proinflammatory cytokines and atherosclerotic effects of a chronic infection mimicked by lipopolysaccharide. AB - OBJECTIVE: Expression of human apolipoprotein (h-apo) A-IV in apoE-deficient (apoE(0)) mice (h-apoA-IV/E(0)) reduces susceptibility to atherosclerosis. Chronic infection mimicked by exposure to lipopolysaccharide (LPS) increases the size of atherosclerosis lesions in apoE(0) mice. Thus, we used h-apoA-IV/E(0) mice to determine whether h-apoA-IV plays a protective role after LPS administration. METHODS AND RESULTS: We injected apoE(0), h-apoA-IV/E(0), and C57Bl/6 (wild-type) mice intraperitoneally with either LPS or phosphate-buffered saline (PBS) every week for 10 weeks. Atherosclerotic lesions were significantly smaller in h-apoA-IV/E(0) mice treated with LPS than in their apoE(0) counterparts. The titers of IgG2a and IgG2b autoantibodies to oxidized low density lipoprotein (LDL) were higher in the LPS-group of h-apoA-IV/E(0) mice than in apoE(0) mice, suggesting that the Th1 response is stronger in the presence of h-apoA-IV. Lymphocytes from the blood, liver, spleen, and thymus of h apoA-IV/E(0) mice treated with LPS produced less IL-4, INF-gamma, and TNF-alpha proinflammatory cytokines than their apoE(0) counterparts. Furthermore, we demonstrated that recombinant h-apoA-IV blocks the LPS-induced stimulation of monocytes. CONCLUSIONS: The expression of h-apoA-IV in apoE(0) mice reduces the susceptibility to atherogenesis and decreases the secretion of proinflammatory cytokines after LPS administration. PMID- 14751812 TI - Angiotensin-converting enzyme D/I gene polymorphism and age-related changes in pulse pressure in subjects with hypertension. AB - Background- Few studies have examined the possible influence of gene polymorphisms on the increase of systolic blood pressure (SBP) and pulse pressure (PP) with age, although in older populations, SBP>160 mm Hg or PP>60 mm Hg are strong mechanical factors predicting cardiovascular mortality. METHODS AND RESULTS: This cross-sectional study involved 315 men and 154 women with either systolic-diastolic or isolated systolic hypertension. Using polymerase chain reaction, the angiotensin-converting enzyme (ACE) D/I gene polymorphism was investigated separately in men and women, enabling us to determine the relationships between age and PP, SBP, and diastolic blood pressure (DBP) for each genotype in each population. In men, most of which were under 50 years of age, the slope of the age-PP and age-SBP (but not age-DBP) relationships differed significantly between genotypes (P=0.0096 and 0.0175). The interslope difference was unmodified after adjustments involving all of the following factors together: plasma glucose, cholesterol, creatinine, potassium, body weight, tobacco consumption, mean blood pressure, and previous antihypertensive therapy. Adjustment of the two latter parameters alone significantly attenuated the interslope difference. Based on logistic regressions, the DD genotype was shown to independently predict a PP>60 mm Hg but not a SBP>160 mm Hg. CONCLUSIONS: In men, the ACE D/I gene polymorphism independently modulates age-related increase of PP, and potentially modulates the resulting cardiovascular risk. This finding requires the development of long-term follow-up. PMID- 14751813 TI - The "CholesteROR" protective pathway in the vascular system. AB - Retinoic acid receptor-related Orphan Receptor alpha (RORalpha) is a member of the nuclear hormone receptor superfamily. RORalpha has long been considered as a constitutive activator of transcription in the absence of exogenous ligand; however, cholesterol has recently been identified as a natural ligand of RORalpha. The spontaneous staggerer (sg/sg) mutation is a deletion in the Rora gene that prevents the translation of the ligand-binding domain (LBD), leading to the loss of RORalpha activity. The homozygous Rora(sg/sg) mutant mouse, of which the most obvious phenotype is ataxia associated with cerebellar degeneration, also displays a variety of other phenotypes, including several vascular ones; in particular, dysfunction of smooth muscle cells and enhanced susceptibility to atherosclerosis. Moreover, RORalpha appears to participate in the regulation of plasma cholesterol levels, and has been shown to positively regulate apolipoprotein (apo)A-I and apoC-III gene expression. Yet its activity is regulated by cholesterol itself, making RORalpha an intracellular cholesterol target. PMID- 14751814 TI - Evidence for a role of macrophage migration inhibitory factor in vascular disease. AB - OBJECTIVE: Inflammation plays an essential role in atherosclerosis and restenosis. Macrophage migration inhibitory factor (MIF) is a proinflammatory cytokine that is widely expressed in vascular cells. However, there is no in vivo evidence that MIF participates directly in vascular injury and repair. Therefore, we investigated the effect of MIF blockade on the response to experimental angioplasty in atherosclerosis-susceptible mice. METHODS AND RESULTS: Carotid artery dilation (2.5 atm) and complete endothelial denudation were performed in male C57BL/6J LDL receptor-deficient mice treated with a neutralizing anti-MIF or isotype control monoclonal antibody. After 7 days and 28 days, intimal and medial sizes were measured and intima/media area ratio (I/M) was calculated. Intimal thickening and I/M were reduced significantly by anti-MIF compared with control antibody. Vascular injury was accompanied by progressive vessel enlargement or "positive remodeling" that was comparable in both treatment groups. MIF blockade was associated with reduced inflammation and cellular proliferation and increased apoptosis after injury. CONCLUSIONS: Neutralizing MIF bioactivity after experimental angioplasty in atherosclerosis-susceptible mice reduces vascular inflammation, cellular proliferation, and neointimal thickening. Although the molecular mechanisms responsible for these effects are not yet established, these data prompt further research directed at understanding the role of MIF in vascular disease and suggest novel therapeutic interventions for preventing atherosclerosis and restenosis. PMID- 14751815 TI - Subclasses of low-density lipoprotein and very low-density lipoprotein in familial combined hyperlipidemia: relationship to multiple lipoprotein phenotype. AB - OBJECTIVE: The present study addresses the presence of distinct metabolic phenotypes in familial combined hyperlipidemia (FCHL) in relation to small dense low-density lipoprotein (sd LDL) and very low-density lipoprotein (VLDL) subclasses. METHODS AND RESULTS: Hyperlipidemic FCHL relatives (n=72) were analyzed for LDL size by gradient gel electrophoresis. Pattern B LDL (sd LDL, particle size <258 A) and pattern A LDL (buoyant LDL, particle size > or =258 A) were defined. Analyses showed bimodal distribution of LDL size associated with distinct phenotypes. Subjects with predominantly large, buoyant LDL showed a hypercholesterolemic phenotype and the highest apo B levels. Subjects with predominantly sd LDL showed a hypertriglyceridemic, low high-density lipoprotein (HDL) cholesterol phenotype, with moderately elevated apoB, total cholesterol level, and LDL cholesterol level. Subjects with both buoyant LDL and sd LDL (pattern AB, n=7) showed an intermediate phenotype, with high normal plasma triglycerides. VLDL subfraction analysis showed that the sd LDL phenotype was associated with a 10-times higher number of VLDL1 particles of relatively lower apo AI and apo E content, as well as smaller VLDL2 particles, in combination with increased plasma insulin concentration in comparison to pattern A. CONCLUSIONS: The present observations underscore the importance of the VLDL triglyceride metabolic pathway in FCHL as an important determinant of the phenotypic heterogeneity of the disorder. PMID- 14751818 TI - Conceptual "Heat-Driven" approach to the synthesis of DNA oligonucleotides on microarrays. AB - The discovery of deoxyribonucleoside cyclic N-acylphosphoramidites, a novel class of phosphoramidite monomers for solid-phase oligonucleotide synthesis, has led to the development of a number of phosphate protecting groups that can be cleaved from DNA oligonucleotides under thermolytic neutral conditions. These include the 2-(N-formyl-N-methyl)aminoethyl, 4-oxopentyl, 3-(N-tert-butyl)carboxamido-1 propyl, 3-(2-pyridyl)-1-propyl, 2-[N-methyl-N-(2-pyridyl)]aminoethyl, and 4 methythiobutyl groups. When used for 5'-hydroxyl protection of nucleosides, the analogous 1-phenyl-2-[N-methyl-N-(2-pyridyl)]aminoethyloxycarbonyl group exhibited excellent thermolytic properties, which may permit an iterative "heat driven" synthesis of DNA oligonucleotides on microarrays. In this regard, progress has been made toward the use of deoxyribonucleoside cyclic N acylphosphoramidites in solid-phase oligonucleotide syntheses without nucleobase protection. Given that deoxyribonucleoside cyclic N-acylphosphoramidites produce oligonucleotides with heat-sensitive phosphate protecting groups, blocking the 5' hydroxyl of these monomers with, for example, the thermolabile 1-phenyl-2-[N methyl-N-(2-pyridyl)]aminoethyloxycarbonyl group may provide a convenient thermo controlled method for the synthesis of oligonucleotides on microarrays. PMID- 14751819 TI - Reading, writing, and modulating genetic information with boranophosphate mimics of nucleotides, DNA, and RNA. AB - The P-boranophosphates are efficient and near perfect mimics of natural nucleic acids in permitting reading and writing of genetic information with high yield and accuracy. Substitution of a borane (-BH3) group for oxygen in the phosphate ester bond creates an isoelectronic and isosteric mimic of natural nucleotide phosphate esters found in mononucleotides, i.e., AMP and ATP, and in RNA and DNA polynucleotides. Compared to natural nucleic acids, the boranophosphate RNA and DNA analogs demonstrate increased lipophilicity and resistance to endo- and exonucleases, yet they retain negative charge and similar spatial geometry. Borane groups can readily be introduced into the NTP and dNTP nucleic acid monomer precursors to produce alpha-P-borano nucleoside triphosphate analogs (e.g., NTPalphaB and dNTPalphaB). The NTPalphaB and dNTPalphaB are, in fact, good to excellent substrates for RNA and DNA polymerases, respectively, and allow ready enzymatic synthesis of RNA and DNA with P-boranophosphate linkages. Further, boranophosphate polymer products are good templates for replication, transcription, and gene expression; boronated RNA products are also suitable for reverse transcription to cDNA. Fully substituted boranophosphate DNA can activate the RNase H cleavage of RNA in RNA:DNA hybrids. Moreover, certain dideoxy NTPalphaB analogs appear to be better substrates for viral reverse transcriptases than the regular ddNTPs, and may offer promising prodrug alternatives in antiviral therapy. These properties make boranophosphates promising candidates for diagnostics; aptamer selection; gene therapy; and antiviral, antisense, and RNAi therapeutics. The boranophosphates constitute a versatile family of phosphate mimics for processing genetic information and modulating gene function. PMID- 14751820 TI - Modulation of Toll-like Receptor 9 Responses through Synthetic Immunostimulatory Motifs of DNA. AB - Bacterial, plasmid, and synthetic DNA containing unmethylated CpG dinucleotides in specific sequence contexts activate the vertebrate innate immune system. A pattern recognition receptor (PRR), toll-like receptor 9 (TLR9), recognizes CpG DNA and activates signaling cascade leading to the secretion of a number of cytokines and chemokines. Our extensive structure-immunostimulatory activity relationship studies showed that a number of synthetic pyrimidine (Y) and purine (R) nucleotides are accepted by the receptor as substitutes for natural deoxycytidine and deoxyguanosine in a CpG dinucleotide. These studies permitted development of synthetic immunostimulatory motifs YpG, CpR, and YpR and established the nucleotide motif recognition pattern of the receptor. A number of site-specific chemical modifications in the flanking sequences to the CpG dinucleotide permitted modulation of immunostimulatory affects in a predictable manner. Our studies also showed that TLR9 recognizes and reads the CpG DNA sequence from the 5'-end. Design of oligonucleotides with two 5'-ends, immunomers, resulted in potent immunomodulatory agents with distinct cytokine profiles. Immunomers containing synthetic immunostimulatory motifs produced different cytokine induction profiles compared with natural CpG motifs. Importantly, some of these synthetic motifs showed optimal activity in both mouse and human systems without requiring to change sequences, suggesting overriding the species-dependent specificity of the receptor by the use of synthetic motifs. In this article, we review current understanding of structural recognition and functional modulation of TLR9 receptor by second-generation immunomodulatory oligonucleotides and their potential application as wide spectrum therapeutic agents. PMID- 14751821 TI - The effect of hydroxyurea and trichostatin a on targeted nucleotide exchange in yeast and Mammalian cells. AB - Targeted nucleotide exchange (TNE) is a process by which a synthetic DNA oligonucleotide, partially complementary to a site in a chromosomal or an episomal gene directs the reversal of a single nucleotide at a specific site. To protect against nuclease digestion, the oligonucleotide is modified with derivative linkages among the terminal bases. We have termed these molecules modified single-stranded oligonucleotides (MSOs). Current models suggest that the reaction occurs in two steps. The first, DNA pairing, involves the alignment of the MSO with the target site and its assimilation into the target helix forming a D-loop. The second phase centers around the repair of a single base mismatch formed between the MSO and its complementary strand in the D-loop. Nucleotide exchange is promoted in all likelihood by the mismatch repair system. A critical feature of successful TNE is the accessibility of the target site for the MSO and the factors that increase the dynamic nature of the chromatin that will likely increase the frequency. Here, we report that two factors, trichostatin A and hydroxyurea, elevate gene repair of a mutant hygromycin gene in Saccharomyces cerevisiae and a mutant eGFP gene in a mammalian cell line, MCF-10AT1 cells. Trichostatin A (TSA) acts by preventing the deacetylation of histones while hydroxyurea (HU) reduces the rate of replication. Both of these activities, by their very nature, create a more open configuration of the MSO into the target site. PMID- 14751822 TI - Short interfering RNA (siRNA)-mediated RNA interference (RNAi) in human cells. AB - Transient gene silencing in mammalian cells can be mediated by double stranded RNA (dsRNAs) molecules of approximately 20-25 nucleotides termed short interfering (siRNAs). Naturally occurring siRNAs in lower eukaryotes have characteristic structural elements; however, little is known about what features are critical for an exogenous siRNA to mediate RNAi in mammals. We have recently determined some of the critical parameters that influence the efficiency of siRNA mediated RNAi in mammalian cells and have been considering the use of RNAi as a functional genomics tool, particularly for high throughput analysis, and the potential use of RNAi as a therapeutic tool. PMID- 14751823 TI - RNA-mediated inhibition of HIV in a gene therapy setting. AB - At present, treatment for HIV-1 infection employs highly active anti-retroviral therapy (HAART), which utilizes a combination of RT and protease inhibitors. Unfortunately, HIV can escape many therapies because of its high mutation rate and the complexity of its pathogenesis. HIV-1 integrates into the cellular genome, which facilitates persistence and acts as a reservoir for reactivation and replication. As an alternative or adjuvant to chemotherapy we have been developing an RNA-based gene therapy approach for the treatment of HIV-1 infection. This article summarizes the various RNA based technologies that we have developed for potential application in a gene therapy setting. PMID- 14751824 TI - Therapeutic potentialities of EWS-Fli-1 mRNA-targeted vectorized antisense oligonucleotides. AB - We have used structured antisense oligonucleotides (AON), which are protected against extra and intracellular degradation by their internal structure. We have shown that if correctly designed this structure does not prevent them from hybridizing to the mRNA target. This concept allows reducing the number of thioate groups in the oligonucleotide and therefore the potential toxicity. Junction oncogenes are found in cancers such as certain leukemias, Ewing sarcoma, and thyroid papillary carcinomas. Ewing sarcoma is a cancer of children and young adults with bone metastasis. It is caused by a chromosomic translocation t(11;22) (q24;q12) creating a fusion gene between the genes EWS and Fli-1 giving rise to a chimeric protein which is an unnatural transcription factor. Immortalized NIH/3T3 cells transfected by the EWS-Fli-1 cDNA under the control of the LTR retroviral promoter--which do not undergo apoptosis and which became tumoral--were used for this study. As a model of Ewing sarcoma in nude mice, we have used permanently expressing human EWS-Fli-1 cells grafted to nude mice. The nanospheres or nanocapsules have been used to deliver two different AON: a phosphorothioate, and a structured chimeric AON, both targeted toward the junction area of EWS-Fli-1. Both types of AON-loaded nanoparticles inhibited the growth of the xenografted tumor after intratumoral injections into nude mice, whereas similar nanoparticles with control oligonucleotides had no effect. With AON in nanospheres, we have shown after 24 hours that the mRNA of EWS-Fli-1 was specifically down-regulated, confirming the antisense activity of the targeted AON. PMID- 14751825 TI - HER-2-targeted antisense oligonucleotide results in sensitization of head and neck cancer cells to chemotherapeutic agents. AB - Existing HER-2 targeted therapies for human head and neck cancers, usually administered in combination with chemotherapeutic drugs or irradiation, include monoclonal antibodies to HER-2, receptor tyrosine kinase inhibitors and HER-2 specific immunotoxins. Instead of targeting the existing protein, interference with HER-2 mRNA translation by antisense oligonucleotides may be a more efficient method to downregulate levels of HER-2 protein for combination therapy. To test this hypothesis we have used a phosphorothioate pentadecamer, complementary to the HER-2 mRNA initiation codon region (AS HER-2 ODN), to increase sensitivity to four chemotherapeutic agents in human head and neck cancer cell lines, all of which express low levels of the HER-2 protein. To improve delivery into tumor cells, the AS HER-2 ODN was complexed with our previously established folate liposome delivery system. Cell survival assays and Western blot analysis data demonstrated that folate-liposome mediated AS HER-2 oligonucleotide treatment inhibited cell growth and HER-2 expression, and induced apoptosis in SCC-25CP cells. Moreover, there was a synergistic effect on the percent of apoptotic cells. Additionally, the combination of folate-liposome-AS HER-2 ODN and CDDP had a synergistic effect on the induction of apoptosis. Using confocal microscopy, FITC labeled ODN (FITC-ODN) in complex with folate-liganded, rhodamine (Rh) labeled, cationic liposomes was observed to enter SCC-25CP head and neck tumor cells within 3 to 6 h. Intracellularly, the FITC-ODN separated from the Rh-folate liposomes, and FITC-ODN accumulated in the nucleus while Rh-liposomes remained in punctate cytoplasmic structures. Thus, folate-liposome-mediated delivery of AS HER-2 ODN has potential as a new means of increasing the responsiveness of head and neck cancer to conventional chemotherapy. PMID- 14751826 TI - Antisense to apoptosis inhibitors facilitates chemotherapy and TRAIL-induced death signaling. AB - Impaired apoptosis signaling is associated with tumor development and confers resistance to chemotherapy and apoptosis triggered by the extrinsic death receptor pathway including Fas and TRAIL-R1/R2. In addition to genetic and epigenetic alterations, such as mutational inactivation and silencing of potential tumor suppressor genes, the antiapoptotic proteins Bcl-2, Bcl-xL, and survivin are overexpressed in many human tumors, and targeted inhibition of their expression has potential to facilitate apoptosis induced by various stimuli. We have used antisense and RNAi technology to counteract the expression of these antiapoptotic proteins in various tumor cell types and investigated the effect of this intervention on apoptosis induction by chemotherapy and the tumor-selective death ligand TRAIL. The oligonucleotide targeting Bcl-2 and Bcl-xL was used in the 2'-MOE or LNA-modified gapmer format, the survivin siRNA was derived from the sequence of an effective first generation antisense oligonucleotide. Modulation of gene expression was monitored by real-time PCR and Western blotting, cell death was determined in cell growth and apoptosis assays. In the tumor cells tested, downregulation of Bcl-2, Bcl-xL or survivin expression facilitated apoptosis via the intrinsic and extrinsic signaling pathway and sensitized tumor cells to various chemotherapeutic agents and to TRAIL. All combinations of antisense and chemotherapy as well as of Bcl-2/Bcl-xL antisense and TRAIL resulted in more than additive cytotoxicity. Although survivin represents a promising target for antisense therapy owing to its tumor-selective expression, its downregulation less effectively enhanced TRAIL-induced apoptosis compared to the Bcl-2/Bcl-xL antisense approach. Our data suggest the use of Bcl-2-, Bcl-xL- and survivin-directed antisense therapy to improve the treatment options for apoptosis-resistant cancer. PMID- 14751827 TI - Clusterin and IGFBPs as antisense targets in prostate cancer. AB - The primary hurdle to improved survival of advanced prostate cancer is our failure to prevent or treat the tumor's progression to its lethal and untreatable stage of androgen independence. Novel treatment modalities designed to prevent androgen-independent progression including prostate cancer metastasis are required. Accelerated identification and characterization of cancer-relevant molecular targets has sparked considerable interest in the development of new generations of anticancer agents that specifically inhibit a progression-relevant target. Antisense oligonucleotides, short synthetic stretches of chemically modified DNA capable of specifically hybridizing to the mRNA of a chosen cancer relevant target gene, promise to show enhanced specificity for malignant cells with a favorable side-effect profile due to well-defined and tailored modes of action. Although not all of the challenges have been met to date, emerging clinical evidence supports the premise that antisense oligonucleotides stand a realistic chance of emerging as major partners of rationally designed anticancer regimens. The rationale and status of antisense targeting of the treatment resistance factor clusterin and of insulin-like growth factor binding protein (IGFBP) 2 and 5 are discussed. PMID- 14751828 TI - The therapeutic potential of antigen-oligonucleotide conjugates. AB - Conjugation of protein antigen with immunostimulatory oligonucleotides creates a potent immunogen that elicits antigen-specific antibody responses, a Th1-biased cytokine profile, and CD8 cytotoxic T lymphocyte activity. The wide range of humoral and cellular immune responses induced by these conjugates suggests they can be used to create effective vaccines against infectious pathogens and tumors and to beneficially modulate the immune responses seen in allergic diseases. This review summarizes the available data on the use of antigen-oligonucleotide conjugates and discusses their potential use for the treatment of infectious, oncologic, and allergic diseases in humans. PMID- 14751829 TI - Regulation of CpG-induced immune activation by suppressive oligodeoxynucleotides. AB - Bacterial DNA and synthetic oligodeoxynucleotides (ODN) containing unmethylated "CpG motifs" stimulate an innate immune response characterized by the production of cytokines, chemokines, and polyreactive Igs that promote host survival following infectious challenge. Yet CpG-driven immune activation can have deleterious consequences, such as increasing the host's susceptibility to autoimmune disease. The immunomodulatory activity of CpG DNA can be blocked by DNA containing "suppressive" motifs. This work explores the rules governing cellular recognition of stimulatory and suppressive motifs, and the resultant modulation of the immune system. Results suggest that both CpG and suppressive ODN may find use as therapeutic agents. PMID- 14751830 TI - CRE-enhancer DNA decoy: a tumor target-based genetic tool. AB - Enhancer DNA decoy oligonucleotides (ODNs) inhibit transcription by competing for transcription factors. A decoy ODN composed of the cAMP response element (CRE) inhibits CRE-directed gene transcription and tumor growth without affecting normal cell growth. We used DNA microarrays to analyze the global gene expression in tumors exposed to the CRE-decoy ODN. The CRE decoy upregulated the AP-2beta transcription factor gene in tumors but not in the livers of host animals. The upregulated expression of AP-2beta was clustered with other upregulated genes involved in development and cell differentiation. Concomitantly, another cluster of genes involved in cell proliferation and transformation was downregulated. The observed alterations indicate that CRE-directed transcription favors tumor growth. Evidence presented here suggests that the CRE-decoy ODN may provide a target-based genetic tool for treating cancer, viral diseases, and other diseases in which CRE-directed transcription is abnormally used. PMID- 14751831 TI - Antigene radiotherapy: targeted radiodamage with 125i-labeled triplex-forming oligonucleotides. AB - Antigene radiotherapy is based upon damaging selected genes by a high dose of radiation from radionuclides delivered to this gene by a sequence-specific DNA binding molecule. Here we describe our recent trials of antigene radiotherapy using the human mdr1 gene over-expressed in KB-V1 cells as a model. As a delivery molecule, we used a triplex-forming oligonucleotide (TFO) with a binding site in intron 14 of mdr1. This TFO was labeled with an Auger-electron-emitting radionuclide 125I. Decay of 125I releases a shower of low energy electrons that produce DNA strand breaks mostly within 10 bp from the decay site. Targeting in situ was assessed by restriction enzyme digestion of the DNA recovered from the TFO-treated cells followed by Southern hybridization with DNA probes flanking the target sequence. Double-strand breaks in the target sequence were detected in purified nuclei and digitonin-permeabilized cells, but not in the intact cells when TFO were delivered with liposomes. On the basis of these observations we hypothesized that there are cytoplasmic factors that bind such TFO and deliver them into the nucleus, but do not release them inside the nucleus, thus preventing TFO from binding their genomic targets. To test this hypothesis we (i) delivered TFO along with an excess of unlabeled oligonucleotide with an arbitrary sequence ("ballast") and (ii) conjugated TFO with a nuclear localization sequence peptide (NLS). We have found that TFO/NLS conjugates cleaved the target in a concentration-dependent manner regardless of the presence of the "ballast" oligonucleotide. In contrast, TFO without NLS cleaved the target only in the presence of an excess of the "ballast." These results may provide a new insight into the mechanism of intracellular transport of oligonucleotides. PMID- 14751832 TI - Gene targeting by triple helix-forming oligonucleotides. AB - Effective gene targeting reagents would have widespread utility for genomic manipulation including transgenic cell and animal construction and for gene therapy. They would also be useful in basic research as probes of chromatin structure, and as tools for studying the repair and mutagenesis of targeted DNA damage. We are developing triple helix-forming oligonucleotides (TFOs) for gene targeting in living mammalian cells. Challenges to TFO bioactivity include the impediments to the biochemistry of triplex formation presented by the physiological environment and the charge repulsion between the duplex and the third strand. In addition, there are biological constraints to target access imposed by mammalian chromatin structure. Here we describe the oligonucleotide modification format that appears to support biological activity of TFOs. In addition we show that manipulation of the cell biology, specifically the cell cycle, has a dramatic influence on TFO bioactivity. PMID- 14751833 TI - Potent inhibition of HhaI DNA methylase by the aglycon of 2-(1H)-pyrimidinone riboside (zebularine) at the GCGC recognition domain. AB - A short oligodeoxynucleotide (ODN) with 2-(1H)-pyrimidinone at the HhaI DNA methyltransferase target site (GCGC) is shown to induce a level of inhibition of methyl transfer and thermal stability of the complex with the enzyme identical to that achieved with a similar ODN substituted with 5-azacytosine. The drugs responsible for these effects-zebularine and 5-azacytidine/2'-deoxy-5-azacytidine are contrasted in terms of chemical stability and possible metabolic activation by a brief structure-activity analysis. PMID- 14751834 TI - Imaging oncogene expression. AB - In 2003, approximately 39,800 women in the US will die from breast cancer. Mammography and physical examination miss up to 40% of early breast cancers. Moreover, if an abnormality is found, an invasive diagnostic procedure must still be performed to determine if the breast contains atypia or cancer, even though approximately 85% of abnormalities are benign. Scintigraphic imaging of gene expression in vivo by noninvasive means could direct physicians to appropriate targets for intervention at the onset of disease and thereby significantly impact patient management. Until now, no method has been available to image specific overexpressed oncogene mRNAs in vivo by scintigraphic imaging. We hypothesize that gamma-emitting Tc-99m-PNA-peptides can be taken up by human ER+ and ER- breast cancer xenografts, hybridize to complementary mRNA targets in those cells, and concentrate sufficiently in tumor tissue to allow noninvasive imaging of oncogene overexpression. To prepare the probes, peptide analogs of insulin-like growth factor 1 (IGF1) were extended from a solid support by Fmoc coupling. Peptide nucleic acid (PNA) dodecamers antisense to CCND1 and MYC mRNAs were then extended from the N-terminus of IGF1, followed by a chelator peptide, using Fmoc coupling for all residues. The cysteine thiols were cyclized on the solid support, either before or after PNA extension. This simplified synthetic approach allows preparation of a variety of multipeptide disulfide-bridged PNA chimeras. A chelating peptide-PNA chimera antisense to MYC mRNA was then labeled efficiently with Tc-99m, yielding a single product. Tissue distribution studies of antisense and mismatch chimeras at 4 h and 24 h after administration displayed modest accumulation in the liver and kidneys, with appreciable levels in tumors. This result enables testing of Tc-99m-peptide-PNA probes to image gene expression in tumors. PMID- 14751835 TI - Custom cancer therapies: safe and effective treatments for most or all cancers. AB - Gene Tools, LLC has embarked on a program to develop safe and effective custom therapies for essentially all cancers. This entails: 1) identifying in each patient's cancer multiple molecular targets which are absent from that patient's normal cells, but are present in and essential to that patient's cancer; and 2) treating that patient with a custom cocktail of therapeutic agents effective for specifically inhibiting those selected targets in that patient's cancer. PMID- 14751836 TI - Antisense to the early growth response-1 gene (Egr-1) inhibits prostate tumor development in TRAMP mice. AB - Egr-1 is a transcription factor induced by stress or injury, mitogens, and differentiation factors. Egr-1 regulates the expression of genes involved in growth control or survival. Expression of Egr-1 results in either promotion or regression of cell proliferation, depending on cell type and environment. Egr-1 acts as a tumor suppressor in many cell types and loss of Egr-1 has been proposed to contribute to cancer progression. There is strong new evidence however suggesting that Egr-1 overexpression is involved in prostate cancer progression. For example, Egr-1 expression levels are elevated in human prostate carcinomas in proportion to grade and stage. Furthermore, prostate cancer progression was significantly delayed in two models of prostate cancer mice lacking Egr-1. Our objective in the present study is to test whether inhibition of Egr-1 function would block cell proliferation and inhibit the transformed phenotype of prostate cancer cells in vitro and in vivo. We describe the development of high affinity and high specificity antisense oligonucleotides that efficiently inhibit Egr-1 expression. We show that inhibition of Egr-1 expression in mouse or human prostate cancer cells decreased proliferation and reduced the capacity of these cells to form colonies and to grow in soft agar. Conversely, stable expression of Egr-1 in normal human prostate epithelial 267B1 cells promoted transformation. In TRAMP mice, treatment with Egr-1 antisense oligonucleotides delayed the occurrence of prostate tumors. Importantly, Egr-1 antisense showed little or no toxicity when injected into animals. Finally, we identified a few genes such as cyclin D2, p19ink4d, and Fas that are directly regulated by Egr-1 in prostate cancer cells and that control cell cycle and survival. PMID- 14751837 TI - Chemosensitization and radiosensitization of human cancer by antisense anti-MDM2 oligonucleotides: in vitro and in vivo activities and mechanisms. AB - MDM2 oncogene is overexpressed in many human cancers including breast, colon, and prostate cancer, and MDM2 levels are associated with poor prognosis in patients with cancer. Here, we summarize the investigation of the functions of MDM2 oncogene in human cancer growth and the value of MDM2 as a drug target for prostate cancer therapy by using antisense to inhibit MDM2 expression. Antisense anti-human-MDM2 oligonucleotides and mismatch controls were tested in in vitro and in vivo human cancer models for antitumor activity. Targeted gene products and related proteins were analyzed and the antitumor activity was determined when the oligonucleotides were used alone or in combination with cancer chemotherapeutics and radiation therapy. The antisense oligonucleotide specifically inhibited MDM2 expression in a dose- and time-dependent manner, resulting in significant antitumor activity in vitro and in vivo. The antisense oligonucleotides also potentiated the effects of p53 activation and p21 induction by chemotherapeutic agents 10-hydroxycamptothecin, adriamycin, 5-fluorouracil, and paclitaxel. In a dose-dependent manner, the antisense oligonucleotide showed antitumor activity in nude mice bearing human cancer xenografts and increased therapeutic effectiveness of the chemotherapeutic agents irinotecan, paclitaxel, and Rituxan and radiation therapy. These results indicate that MDM2 has a role in various tumor growth through both p53-dependent and p53-independent mechanisms, indicating that MDM2 inhibitors have a broad spectrum of antitumor activities in human cancers regardless of p53 status. These results provide a basis for clinical evaluation of antisense anti-MDM2 oligonucleotides as chemosensitizer and radiosensitizer. PMID- 14751838 TI - Antisense targeting protein kinase A type I as a drug for integrated strategies of cancer therapy. AB - We have studied the role of protein kinase A (PKA) and its type I isoform (PKAI) in the transduction of mitogenic signaling, apoptosis, and angiogenesis. We have contributed to the development of selective inhibitors of PKAI, including a hybrid DNA/RNA mixed backbone oligonucleotide (AS-PKAI). We, and others, have demonstrated that AS-PKAI has a cooperative antitumor effect with a selected class of cytotoxic drugs and with radiotherapy in vitro and in vivo and that these effects can also be obtained following oral adinistration. Previously, we developed a series of therapeutic models based on the pleiotropic role played by PKAI in cell proliferation, apoptosis, and angiogenesis. On the basis of our former demonstration of functional and structural interactions of PKAI and the activated epidermal growth factor receptor (EGFR), we have shown that the combined blockade of both signaling molecules by AS-PKAI and either the monoclonal antibody C225 (erbitux) or the small molecule ZD1839 (gefitinib), results in a marked cooperative antitumor effect in a variety of human tumor models. A further cooperative antitumor effect can be obtained when AS-PKAI is used in combination with both EGFR inhibitors and either cytotoxic drugs or radiotherapy. The antitumor activity is associated with inhibition of growth factors and angiogenic factors production and to induction of apoptosis. In light of the recently demonstrated role of PKAI on the bcl-2-dependent apoptotic pathway, we have recently shown a synergistic antitumor, antiangiogenic, and proapoptotic effect of AS-PKAI in combination with antisense bcl-2 (oblimersen) or with a bispecific bcl-2/bcl-xL second generation antisense. A connection between COX-2, EGFR and PKAI was established, and we demonstrated that the combination of AS-PKAI with gefitinib and a COX-2 inhibitor, all adminstered orally, can result in a potent antitumor and antiangiogenic activity. These studies support the development of AS-PKAI as a novel anticancer agent and suggest its potentially relevant role when integrated with conventional treatments and/or other signaling inhibitors in novel therapeutic strategies. PMID- 14751839 TI - Applying antisense technology: Affinitak and other antisense oligonucleotides in clinical development. AB - Progress in the development of antisense drugs over the last decade has led to the approval of the first such drug--Vitravene for AIDS-related CMV retinitis- and the development of a large number of antisense drugs in clinical trials. Antisense drugs are now being studied in Phase 3 trials for patients with cancer and inflammatory bowel disease. Other antisense drugs are in development for rheumatoid arthritis, other inflammatory conditions, and hepatitis C. Still other antisense drugs are entering clinical trials for treatment of metabolic conditions such as diabetes and hyperlipidemia. These latter applications provide the potential for target effects to be more directly measured in the clinic. Improved antisense chemistry, which will enhance the feasibility of subcutaneous and oral administration of antisense drugs and offer the potential of less frequent dosing, is expected to further expand the opportunities for antisense drug development. PMID- 14751840 TI - Clinical studies in patients with solid tumors using a second-generation antisense oligonucleotide (GEM 231) targeted against protein kinase A type I. AB - GEM 231 is a second-generation antisense oligonucleotide targeted against the RIalpha regulatory subunit of cAMP-dependent protein kinase type I (PKA-I). Excessive expression of PKA-I is associated with cell proliferation and transformation, and increased levels of secreted extracellular PKA (ECPKA) are found in the serum of cancer patients. Preclinical studies have demonstrated single-agent antitumor activity of GEM 231 in a variety of human cancer xenograft models, and additive or synergistic antitumor activity has been observed with taxane and/or camptothecin-based combinations. Based on prior safety (MTD) data demonstrating dose-dependent, reversible, and cumulative transaminitis, and high peak plasma concentration (Cmax)-dependent changes in activated partial thromboplastin time (aPTT) with GEM 231 2-h twice-weekly infusions, an alternative schedule of GEM 231 given as a single agent was evaluated in patients with advanced solid tumors. Fourteen patients (median age approximately 60 yrs) with advanced solid malignancies received a total of 78 weeks of therapy. GEM 231 was infused via a CADD pump at 80 mg/m2/day (d) for 3 d/wk (n = 1), then for 5 d/wk at 80 (n = 3), 120 (n = 8), and 180 mg/m2/d (n = 2). One cycle was defined as 4 weeks of therapy. Apparent dose dependency for the occurrence of transaminitis was readily reversible. At 180 mg/m2/d, 2 of 2 patients had cycle 1 dose-limiting toxicity (DLT) transaminitis. One patient treated at 120 mg/m2/d experienced grade 3 transaminase elevations after 8 weeks of therapy, but when serum transaminase values rapidly improved he resumed treatment at 80 mg/m2/d for 6 weeks until tumor progression was documented. Another patient at 120 mg/m2/d developed grade 3 esophagitis after 3 weeks, limiting further dosing. One patient (lung cancer) demonstrated stable disease for 9 weeks. Overall, plasma aPTT was minimally prolonged and changes were transient, peaked at the end of each infusion, and were not associated with spontaneous bleeding. A constitutive symptom (e.g., low-grade fatigue) was common, cumulative, and reversible following discontinuation of therapy. Serum ECPKA was measured by enzymatic assay and Western blotting from blood drawn at the beginning and end of each infusion. Serum ECPKA levels demonstrated a trend to decline with the treatment. In addition to single agent schedules, combination trials were undertaken to assess safety and possible interaction of GEM 231 with taxanes (paclitaxel, docetaxel), given once every 3 weeks (one cycle). While trials using the 2-h twice-weekly GEM 231 infusions are ongoing, preliminary results from both studies show that it is safe to combine paclitaxel or docetaxel with GEM 231. Overall, it is also feasible to administer GEM 231 in combination with taxane or nontaxane chemotherapy (e.g., camptothecins). Phase I combination studies are currently underway to further explore the clinical, pharmacokinetic, and biologic profile of GEM 231 with chemotherapy. PMID- 14751841 TI - Targeting protein kinase C-alpha (PKC-alpha) in cancer with the phosphorothioate antisense oligonucleotide aprinocarsen. AB - Antisense oligonucleotides (ASOs) offer a novel pharmacological platform to develop highly specific drugs. As shown by the clinical development of aprinocarsen, an ASO directed against protein kinase C-alpha (PKC-alpha), this platform has made a remarkable advance from the bench to the bedside. This review summarizes the rationale of the early development of aprinocarsen and current clinical experience. PMID- 14751844 TI - Metabolic and mitogenic effects of IGF-I and insulin on muscle cells of rainbow trout. AB - The relative function of IGF-I and insulin on fish muscle metabolism and growth has been investigated by the isolation and culture at different stages (myoblasts at day 1, myocytes at day 4, and myotubes at day 10) of rainbow trout muscle cells. This in vitro model avoids interactions with endogenous peptides, which could interfere with the muscle response. In these cells, the effects of IGF-I and insulin on cell proliferation, 2-deoxyglucose (2-DG), and l-alanine uptake at different development stages, and the use of inhibitors were studied and quantified. Insulin (10-1,000 nM) and IGF-I (10-100 nM) stimulated 2-DG uptake in trout myocytes at day 4 in a similar manner (maximum of 124% for insulin and of 142% for IGF-I), and this stimulation increased when cells differentiated to myotubes (maximum for IGF-I of 193%). When incubating the cells with PD-98059 and especially cytochalasin B, a reduction in 2-DG uptake was observed, suggesting that glucose transport takes place through specific facilitative transporters. IGF-I (1-100 nM) stimulated the l-alanine uptake in myocytes at day 4 (maximum of 239%), reaching higher values of stimulation than insulin (100-1,000 nM) (maximum of 160%). This stimulation decreased when cells developed to myotubes at day 10 (118% for IGF-I and 114% for insulin). IGF-I (0.125-25 nM) had a significant effect on myoblast proliferation, measured by thymidine incorporation (maximum of 170%), and required the presence of 2-5% fetal serum (FBS) to promote thymidine uptake. On the other hand, insulin was totally ineffective in stimulating thymidine uptake. We conclude that IGF-I is more effective than insulin in stimulating glucose and alanine uptake in rainbow trout myosatellite cells and that the degree of stimulation changes when cells differentiate to myotubes. IGF I stimulates cell proliferation in this model of muscle in vitro and insulin does not. These results indicate the important role of IGF-I on growth and metabolism of fish muscle. PMID- 14751843 TI - The M2 muscarinic receptor mediates in vitro bladder contractions from patients with neurogenic bladder dysfunction. AB - Bladder muscle specimens from seven patients with neurogenic bladder dysfunction were analyzed to determine whether the muscarinic receptor subtype mediating contraction shifts from M(3) to the M(2) subtype as found in the denervated, hypertrophied rat bladder. Seven bladder specimens were analyzed from six female and one male patients. Six of the patients had traumatic cervical spinal cord injuries (C(4)-C(7)), and the other patient had an L(1) congenital myelomeningocele. This was compared with results from bladder specimens obtained from eight organ transplant donors. The affinities of three subtype-selective muscarinic receptor antagonists for inhibition of carbachol-induced contractions were determined. The affinity of the M(3) selective antagonists darifenacin or p fluoro-hexahydrosiladifenadol (p-F-HHSiD) was determined in six of the seven spinal injury patient specimens. The affinity was consistent with M(2)-mediated contractions in four of these six specimens, intermediate between M(2) and M(3) in one specimen, and within the M(3) range in one specimen. The other specimen, tested only with the M(2) selective antagonist methoctramine, showed an M(3) affinity. In the organ donors, the affinity of p-F-HHSiD was within the M(2) range for six of seven specimens, whereas the affinity of darifenacin was within the M(3) range for five of six and intermediate between M(2) and M(3) for the other specimen tested. The affinity of methoctramine in both organ donor specimens tested was within the M(3) range. Whereas normal detrusor contractions are mediated by the M(3) receptor subtype, in patients with neurogenic bladder dysfunction as well as certain organ transplant donors, contractions can be mediated by the M(2) muscarinic receptor subtype. PMID- 14751845 TI - The mechanism of EDHF-mediated responses in subcutaneous small arteries from healthy pregnant women. AB - We studied the importance of endothelium-derived hyperpolarizing factor (EDHF) vs. nitric oxide (NO) and prostacyclin (PGI(2)) in bradykinin (BK)-induced relaxation in isolated small subcutaneous arteries from normal pregnant women. We also explored the contribution of cytochrome P-450 (CYP450) product of arachidonic acid (AA) metabolism, hydrogen peroxide (H(2)O(2)), and gap junctions that have been suggested to be involved in EDHF-mediated responses. Isolated arteries obtained from subcutaneous fat biopsies of normal pregnant women (n = 30) undergoing planned cesarean section were mounted in a wire-myography system. In norepinephrine-constricted vessels, incubation with N(G)-nitro-L-arginine methyl ester (L-NAME) resulted in a significant reduction in relaxation to BK. Simultaneous incubation with L-NAME and indomethacin failed to modify this response further. BK-mediated dilatation in the presence of K(+)-modified solution was decreased to similar level as obtained after incubation with L-NAME. Incubation with L-NAME abolished BK-induced responses in K(+)-modified solution. Sulfaphenazole, a specific inhibitor of CYP450 epoxygenase, and catalase (an enzyme that decomposes H(2)O(2)) did not affect the EDHF-mediated relaxation because concentration-response curves to BK were similar in arteries after incubation with L-NAME vs. L-NAME + sulfaphenazole and L-NAME + catalase. The inhibitor of gap junctions, 18 alpha-glycyrrhetinic acid, significantly reduced BK-mediated relaxation both without and with incubation with L-NAME. We found that both NO and EDHF, but not PGI(2), are involved in the endothelium-dependent dilatation to BK. BK-induced relaxation is almost equally mediated by NO and EDHF. CYP450 epoxygenase metabolites of AA or H(2)O(2) do not account for EDHF mediated response; however, gap junctions are involved in the EDHF-mediated responses to BK in subcutaneous small arteries in normal pregnancy. PMID- 14751846 TI - Stimulation of cardiac sympathetic nerve activity by central angiotensinergic mechanisms in conscious sheep. AB - Central actions of angiotensin play an important role in cardiovascular control and have been implicated in the pathogenesis of hypertension and heart failure. One feature of centrally or peripherally administered angiotensin is that the bradycardia in response to an acute pressor effect is blunted. It is unknown whether after central angiotensin this is due partly to increased cardiac sympathetic nerve activity (CSNA). We recorded CSNA and arterial pressure in conscious sheep, at least 3 days after electrode implantation. The effects of intracerebroventricular infusions of ANG II (3 nmol/h for 30 min) and artificial cerebrospinal fluid (CSF) (1 ml/h) were determined. The response to intracerebroventricular hypertonic saline (0.6 M NaCl in CSF at 1 ml/h) was examined as there is evidence that hypertonic saline acts via angiotensinergic pathways. Intracerebroventricular angiotensin increased CSNA by 23 +/- 7% (P < 0.001) and mean arterial pressure (MAP) by 7.6 +/- 1.2 mmHg (P < 0.001) but did not significantly change heart rate (n = 5). During intracerebroventricular ANG II the reflex relation between CSNA and diastolic blood pressure was significantly shifted to the right (P < 0.01). Intracerebroventricular hypertonic saline increased CSNA (+9.4 +/- 6.6%, P < 0.05) and MAP but did not alter heart rate. The responses to angiotensin and hypertonic saline were prevented by intracerebroventricular losartan (1 mg/h). In conclusion, in conscious sheep angiotensin acts within the brain to increase CSNA, despite increased MAP. The increase in CSNA may account partly for the lack of bradycardia in response to the increased arterial pressure. The responses to angiotensin and hypertonic saline were losartan sensitive, indicating they were mediated by angiotensin AT-1 receptors. PMID- 14751847 TI - Adrenomedullin modulates hemodynamic and cardiac effects of angiotensin II in conscious rats. AB - We examined whether adrenomedullin, a vasoactive peptide expressed in the heart, modulates the increase in blood pressure, changes in systolic and diastolic function, and left ventricular hypertrophy produced by long-term administration of ANG II or norepinephrine in rats. Subcutaneous administration of adrenomedullin (1.5 microg.kg(-1).h(-1)) for 1 wk inhibited the ANG II-induced (33.3 microg.kg(-1).h(-1) sc) increase in mean arterial pressure by 67% (P < 0.001) but had no effect of norepinephrine-induced (300 microg.kg(-1).h(-1) sc) hypertension. Adrenomedullin enhanced the ANG II-induced improvement in systolic function, resulting in a further 9% increase (P < 0.01) in the left ventricular ejection fraction and 19% increase (P < 0.05) in the left ventricular fractional shortening measured by echocardiography, meanwhile norepinephrine-induced changes in systolic function were remained unaffected. Adrenomedullin had no effect on ANG II- or norepinephrine-induced left ventricular hypertrophy or expression of hypertrophy-associated genes, including contractile protein and natriuretic peptide genes. The present study shows that adrenomedullin selectively suppressed the increase in blood pressure and augmented the improvement of systolic function induced by ANG II. Because adrenomedullin had no effects on ANG II- and norepinephrine-induced left ventricular hypertrophy, circulating adrenomedullin appears to act mainly as a regulator of vascular tone and cardiac function. PMID- 14751848 TI - 5-HT induces an arachidonic acid-sensitive calcium influx in rat small intrapulmonary artery. AB - 5-Hydroxytryptamine (5-HT) is a potent pulmonary vasoconstrictor and contributes to hypoxic pulmonary vasoconstriction and pulmonary arterial hypertension. Small intrapulmonary vessels are very sensitive to hypoxia and play a major role for blood flow regulation in the lung. Thus we have investigated the mechanisms involved in the calcium signal to 5-HT in rat small intrapulmonary artery (IPA). Effects of 5-HT were examined in isolated IPA (external diameter <250 microm) from rat. Digital imaging with fura-PE3 was used to record intracellular calcium concentration ([Ca(2+)](i)) and to follow external diameter of the vessels. 5-HT induced a sustained [Ca(2+)](i) variation that was sensitive to the inhibitor of the 5-HT(2A) receptors, ketanserin, and insensitive to voltage-dependent L-type calcium channel blockers (nitrendipine and nicardipine) or voltage-independent calcium channel antagonists (LOE-908, SKF-96365, and gadolinium). The calcium response to 5-HT was also not modified by a sarcoplasmic reticulum Ca(2+)-ATPase inhibitor (cyclopiazonic acid; CPA), which depletes intracellular calcium stores. CPA alone activated a capacitative calcium channel that was sensitive to LOE-908 and insensitive to SKF-96365 and gadolinium. The sustained calcium signal to 5-HT was partly blocked by inhibitors of arachidonic acid production (RHC-80267 and isotetrandrine) and mimicked by application of exogenous arachidonic acid. These results suggest that activation of a noncapacitative, arachidonic acid-sensitive, receptor-operated calcium channel contributes to 5-HT-induced sustained calcium increase in small IPA. PMID- 14751849 TI - Increased pulmonary blood flow does not alter surfactant protein gene expression in lambs within the first week of life. AB - Neonates and infants with congenital heart disease with increased pulmonary blood flow suffer morbidity from poor oxygenation and decreased lung compliance. In a previous experiment involving 4-wk-old lambs with pulmonary hypertension secondary to increased pulmonary blood flow following an in utero placement of an aortopulmonary vascular graft, we found a decrease in surfactant protein (SP)-A gene expression as well as a decrease in SP-A and SP-B protein contents. To determine the timing of these changes, the objective of the present study was to characterize the effect of increased pulmonary blood flow and pulmonary hypertension on SP-A, -B, and -C gene expressions and protein contents within the first week of life. Of eight fetal lambs that underwent the in utero placement of the shunt, there was no difference in the expression of SP-A, -B, and -C mRNA levels or SP-A and -B protein contents compared with age-matched controls. The results showed that, in this model of congenital heart disease with pulmonary hypertension and increased pulmonary blood flow, the effect of the shunt on SP gene expression and protein content was not apparent within the first week of life. PMID- 14751850 TI - Electron microscopic study of actin polymerization in airway smooth muscle. AB - Actin polymerization as part of the normal smooth muscle response to various stimuli has been reported. The actin dynamics are believed to be necessary for cytoskeletal remodeling in smooth muscle in its adaptation to external stress and strain and for maintenance of optimal contractility. We have shown in our previous studies in airway smooth muscle that myosins polymerized in response to contractile activation as well as to adaptation at longer cell lengths. We postulated that the same response could be elicited from actins under the same conditions. In the present study, actin filament formation was quantified electron microscopically in cell cross sections. Nanometer resolution allowed us to examine regional distribution of filaments in a cell cross section. Airway smooth muscle bundles were fixed in relaxed and activated states at two lengths; muscle preparations were also fixed after a period of oscillatory strain, a condition known to cause depolymerization of myosin filaments. The results indicate that contractile activation and increased cell length nonsynergistically enhanced actin polymerization; the extent of actin polymerization was substantially less than that of myosin polymerization. Oscillatory strain increased thin filament formation. Although thin filament density was found higher in cytoplasmic areas near dense bodies, contractile activation did not preferentially enhance actin polymerization in these areas. It is concluded that actin thin filaments are dynamic structures whose length and number are regulated by the cell in response to changes in extracellular environment and that polymerization and depolymerization of thin filaments occur uniformly across the whole cell cross section. PMID- 14751851 TI - Signal transduction events involved in TPA downregulation of SP-A gene expression. AB - Surfactant protein A (SP-A), the most abundant pulmonary surfactant protein, plays a role in innate host defense and blocks the inhibitory effects of serum proteins on surfactant surface tension-lowering properties. SP-A mRNA and protein are downregulated by phorbol esters (TPA) via inhibition of gene transcription. We evaluated the TPA signaling pathways involved in SP-A inhibition in a lung cell line, H441 cells. TPA caused sustained phosphorylation of p44/42 mitogen activated protein kinase (MAPK), p38 MAPK, and c-Jun-NH(2)-terminal kinase. An inhibitor of conventional and novel isoforms of protein kinase C (PKC) and two inhibitors of p44/42 MAPK kinase partially or completely blocked the inhibitory effects of TPA on SP-A mRNA levels. In contrast, inhibitors of conventional PKC alpha and -beta, stress-activated protein kinases, protein phosphatases, protein kinase A, and the phosphatidylinositol 3-kinase pathway had no effect on the TPA mediated inhibition of SP-A mRNA. TPA also stimulated the synthesis of c-Jun mRNA and protein in a time-dependent manner. Inhibitors of the p44/42 MAPK signaling pathway and PKC blocked the TPA-mediated phosphorylation of p44/42 MAPK and the increase in c-Jun mRNA. We conclude that TPA inhibits SP-A gene expression via novel isoforms of PKC, the p44/42 MAPK pathway, and the activator protein-1 complex. PMID- 14751853 TI - Effect of acute global ischemia on the upper limit of vulnerability: a simulation study. AB - The goal of this modeling research is to provide mechanistic insight into the effect of altered membrane kinetics associated with 5-12 min of acute global ischemia on the upper limit of cardiac vulnerability (ULV) to electric shocks. We simulate electrical activity in a finite-element bidomain model of a 4-mm-thick slice through the canine ventricles that incorporates realistic geometry and fiber architecture. Global acute ischemia is represented by changes in membrane dynamics due to hyperkalemia, acidosis, and hypoxia. Two stages of acute ischemia are simulated corresponding to 5-7 min (stage 1) and 10-12 min (stage 2) after the onset of ischemia. Monophasic shocks are delivered in normoxia and ischemia over a range of coupling intervals, and their outcomes are examined to determine the highest shock strength that resulted in induction of reentrant arrhythmia. Our results demonstrate that acute ischemia stage 1 results in ULV reduction to 0.8A from its normoxic value of 1.4A. In contrast, no arrhythmia is induced regardless of shock strength in acute ischemia stage 2. An investigation of mechanisms underlying this behavior revealed that decreased postshock refractoriness resulting mainly from 1) ischemic electrophysiological substrate and 2) decrease in the extent of areas positively-polarized by the shock is responsible for the change in ULV during stage 1. In contrast, conduction failure is the main cause for the lack of vulnerability in acute ischemia stage 2. The insight provided by this study furthers our understanding of mechanisms by which acute ischemia-induced changes at the ionic level modulate cardiac vulnerability to electric shocks. PMID- 14751852 TI - Hepatoma-derived growth factor is a pulmonary endothelial cell-expressed angiogenic factor. AB - Hepatoma-derived growth factor (HDGF) was previously identified as a developmentally regulated cardiovascular and renal gene that is mitogenic for vascular smooth muscle and aortic endothelial cells. As reciprocal interactions of smooth muscle and endothelial cells are necessary for vascular formation, we examined whether HDGF plays a role in angiogenesis. According to immunohistochemistry, HDGF was highly expressed in endothelial cells of nonmuscularized, forming blood vessels of the fetal lung. HDGF was also expressed in endothelial cells of small (20 microm) mature arteries and veins. By Western immunoblotting, HDGF was highly expressed by human pulmonary microvascular endothelial cells in vitro. Adenoviral overexpression of HDGF was mitogenic for human pulmonary microvascular endothelial cells in serum-free medium, stimulating a 1.75-fold increase in bromodeoxyuridine (BrdU) uptake and a twofold increase in cell migration. With the chick chorioallantoic membrane (CAM), a biologic assay for angiogenesis, exogenous recombinant HDGF significantly stimulated blood vessel formation and a dose-dependent reorganization of cells within the CAM into a more compact, linear alignment reminiscent of tube formation. According to double immunostaining for endothelial cells with a transforming growth factor betaII receptor antibody and BrdU as a marker of cell proliferation, exogenous HDGF selectively stimulated endothelial cell BrdU uptake. HDGF also activated specific ERK1/2 signaling and did not overlap with VEGF SAPK/JNK, Akt-mediated pathways. We conclude that HDGF is a highly expressed vascular endothelial cell protein in vivo and is a potent endothelial mitogen and regulator of endothelial cell migration by mechanisms distinct from VEGF. PMID- 14751854 TI - Cerebral artery reactivity changes during pregnancy and the postpartum period: a role in eclampsia? AB - Eclampsia is thought to be similar to hypertensive encephalopathy, whereby acute elevations in intravascular pressure cause forced dilatation (FD) of intrinsic myogenic tone of cerebral arteries and arterioles, decreased cerebrovascular resistance, and hyperperfusion. In the present study, we tested the hypothesis that pregnancy and/or the postpartum period predispose cerebral arteries to FD by diminishing pressure-induced myogenic activity. We compared the reactivity to pressure (myogenic activity) as well as factors that modulate the level of tone of third-order branches (<200 microm) of the posterior cerebral artery (PCA) that were isolated from nonpregnant (NP, n = 7), late-pregnant (LP, 19 days, n = 10), and postpartum (PP, 3 days, n = 8) Sprague-Dawley rats under pressurized conditions. PCAs from all groups of animals developed spontaneous tone within the myogenic pressure range (50-150 mmHg) and constricted arteries at 100 mmHg (NP, 30 +/- 3; LP, 39 +/- 4; and PP, 42 +/- 7%; P > 0.05). This level of myogenic activity was maintained in the NP arteries at all pressures; however, both LP and PP arteries dilated at considerably lower pressures compared with NP, which lowered the pressure at which FD occurred from >175 for NP to 146 +/- 6.5 mmHg for LP (P < 0.01 vs. NP) and 162 +/- 7.7 mmHg for PP (P < 0.01 vs. NP). The amount of myogenic tone was also significantly diminished at 175 mmHg compared with NP: percent tone for NP, LP, and PP animals were 35 +/- 2, 11 +/- 3 (P < 0.01 vs. NP), and 20 +/- 7% (P < 0.01 vs. NP), respectively. Inhibition of nitric oxide (NO) with 0.1 mM N(omega)-nitro-l-arginine (l-NNA) caused constriction of all vessel types that was significantly increased in the PP arteries, which demonstrates significant basal NO production. Reactivity to 5-hydroxytryptamine (serotonin) was assessed in the presence of l-NNA and indomethacin. There was a differential response to serotonin: PCAs from NP animals dilated, whereas LP and PP arteries constricted. These results suggest that both pregnancy and the postpartum period predispose the cerebral circulation to FD at lower pressures, a response that may lower cerebrovascular resistance and promote hyperperfusion when blood pressure is elevated, as occurs during eclampsia. PMID- 14751855 TI - Nitric oxide attenuates insulin- or IGF-I-stimulated aortic smooth muscle cell motility by decreasing H2O2 levels: essential role of cGMP. AB - Insulin and insulin-like growth factor I (IGF-I) both play important roles in vascular remodeling. Moreover, nitric oxide (NO) is well established as a counterregulatory agent that opposes the actions of several vascular agonists, in part by decreasing smooth muscle motility. We tested the hypothesis that NO blocks insulin or IGF-I-induced rat aortic smooth muscle cell motility via a mechanism involving the attenuation of agonist-induced elevation of hydrogen peroxide levels and cGMP as mediator. Insulin or IGF-I induced an increase of hydrogen peroxide levels and cell motility. Both effects were blocked by catalase or diphenyleneiodonium, indicating that hydrogen peroxide elevation is necessary for induction of cell motility. Two NO donors mimicked the effects of catalase, indicating that NO decreases cell motility by suppressing agonist-induced elevation of hydrogen peroxide. A cGMP analogue mimicked the effect of NO, whereas a guanyl cyclase inhibitor blocked the effect of NO on hydrogen peroxide levels, indicating that elevation of cGMP is both necessary and sufficient to account for the reduction of hydrogen peroxide levels. A NO donor as well as a cGMP analogue attenuated insulin-stimulated NADPH activity, indicating that NO decreases hydrogen peroxide levels by inhibiting the generation of superoxide, via a cGMP-mediated mechanism. Finally, exogenous hydrogen peroxide increased cell motility and reversed the inhibitory effect of cGMP. These results support the view that NO plays an antioxidant role via reduction of hydrogen peroxide in cultured rat aortic smooth muscle cells and that this effect is both necessary and sufficient to account for its capacity to decrease cell motility. PMID- 14751856 TI - Limited transfer of cytosolic NADH into mitochondria at high cardiac workload. AB - Glycolysis supplements energy synthesis at high cardiac workloads, producing not only ATP but also cytosolic NADH and pyruvate for oxidative ATP synthesis. Despite adequate Po(2), speculation exists that not all cytosolic NADH is oxidized by the mitochondria, leading to lactate production. In this study, we elucidate the mechanism for limited cytosolic NADH oxidation and increased lactate production at high workload despite adequate myocardial blood flow and oxygenation. Reducing equivalents from glycolysis enter mitochondria via exchange of mitochondrial alpha-ketoglutarate (alpha-KG) for cytosolic malate. This exchange was monitored at baseline and at high workloads by comparing (13)C enrichment between the products of alpha-KG oxidation (succinate) and alpha-KG efflux from mitochondria (glutamate). Under general anesthesia, a left thoracotomy was performed on 14 dogs and [2-(13)C]acetate was infused into the left anterior descending artery for 40 min. The rate-pressure product was 9,035 +/- 1,972 and 21,659 +/- 5,266 mmHg.beats.min(-1) (n = 7) at baseline (n = 7) and with dobutamine, respectively. (13)C enrichment of succinate was 57 +/- 10% at baseline and 45 +/- 13% at elevated workload (not significant), confirming oxidation of [2-(13)C]acetate. However, cytosolic glutamate enrichment, a marker of cytosolic NADH transfer to mitochondria, was dramatically reduced at high cardiac workload (11 +/- 1%) vs. baseline (50 +/- 14%, P < 0.05). This reduced exchange of (13)C from alpha-KG to cytosolic glutamate at high work indicates reduced shuttling of cytosolic reducing equivalents into the mitochondria. Myocardial tissue lactate increased 78%, countering this reduced oxidation of cytosolic NADH. The findings elucidate a contributing mechanism to glycolysis outpacing glucose oxidation in the absence of myocardial ischemia. PMID- 14751857 TI - Red blood cell dysfunction in septic glucose-6-phosphate dehydrogenase-deficient mice. AB - Glucose-6-phosphate dehydrogenase (G-6-PDH) deficiency is the most common known human genetic polymorphism. This study tested the hypothesis that G-6-PDH deficiency worsens sepsis-induced erythrocyte dysfunction. Sepsis (24 h) was induced by cecal ligation and puncture in wild-type (WT) and G-6-PDH-deficient (G 6-PDH activity 15% of WT) mice. Erythrocyte responses were tested in whole blood as well as in subpopulations of circulating erythrocytes. Whereas erythrocyte deformability was similar in unchallenged deficient and WT animals, sepsis decreased erythrocyte deformability that was more pronounced in deficient than WT animals. Sepsis also resulted in anemia and hemolysis in deficient compared with WT animals. Mean corpuscular hemoglobin content and erythrocyte deformability decreased in younger erythrocyte subpopulations from septic deficient compared with WT animals. Sepsis decreased the reduced-to-oxidized glutathione ratio in erythrocytes from both deficient and WT animals; however, plasma glutathione increased more in deficient than in WT animals. Erythrocyte content of band 3 associated with the cytoskeleton was elevated in deficient compared with WT erythrocytes. The antioxidant N-acetyl-l-cysteine in vivo alleviated the sepsis induced decrease in erythrocyte deformability in deficient animals compared with sham-operated control animals. This study demonstrates that a mild degree of G-6 PDH deficiency (comparable to the human class III G-6-PDH deficiencies) worsens erythrocyte dysfunction during sepsis. Increased erythrocyte rigidity and tendency for hemolysis together with alterations in band 3-spectrin interactions may contribute to the immunomodulatory effects of G-6-PDH deficiency observed after major trauma and infections in humans. PMID- 14751858 TI - Transmural mechanics at left ventricular epicardial pacing site. AB - Left ventricular (LV) epicardial pacing acutely reduces wall thickening at the pacing site. Because LV epicardial pacing also reduces transverse shear deformation, which is related to myocardial sheet shear, we hypothesized that impaired end-systolic wall thickening at the pacing site is due to reduction in myocardial sheet shear deformation, resulting in a reduced contribution of sheet shear to wall thickening. We also hypothesized that epicardial pacing would reverse the transmural mechanical activation sequence and thereby mitigate normal transmural deformation. To test these hypotheses, we investigated the effects of LV epicardial pacing on transmural fiber-sheet mechanics by determining three dimensional finite deformation during normal atrioventricular conduction and LV epicardial pacing in the anterior wall of normal dog hearts in vivo. Our measurements indicate that impaired end-systolic wall thickening at the pacing site was not due to selective reduction of sheet shear, but rather resulted from overall depression of fiber-sheet deformation, and relative contributions of sheet strains to wall thickening were maintained. These findings suggest lack of effective end-systolic myocardial deformation at the pacing site, most likely because the pacing site initiates contraction significantly earlier than the rest of the ventricle. Epicardial pacing also induced reversal of the transmural mechanical activation sequence, which depressed sheet extension and wall thickening early in the cardiac cycle, whereas transverse shear and sheet shear deformation were not affected. These findings suggest that normal sheet extension and wall thickening immediately after activation may require normal transmural activation sequence, whereas sheet shear deformation may be determined by local anatomy. PMID- 14751859 TI - Left atrial conduit volume is generated by deviation from the constant-volume state of the left heart: a combined MRI-echocardiographic study. AB - Although modeling the four-chambered heart as a constant-volume pump successfully predicts causal physiological relationships between cardiac indexes previously deemed unrelated, the real four-chambered heart slightly deviates from the constant-volume state by ventricular end systole. This deviation has consequences that affect chamber function, specifically, left atrial (LA) function. LA attributes have been characterized as booster pump, reservoir, and conduit functions, yet characterization of their temporal occurrence or their causal relationship to global heart function has been lacking. We investigated LA function in the context of the constant-volume attribute of the left heart in 10 normal subjects using cardiac magnetic resonance imaging (MRI) and contemporaneous Doppler echocardiography synchronized via ECG. Left ventricular (LV) and LA volumes as a function of time were determined via MRI. Transmitral flow, pulmonary vein (PV) flow, and lateral mitral annular velocity were recorded via echocardiography. The relationship between the MRI-determined diastolic LA conduit-volume (LACV) filling rate and systolic LA filling rate correlate well with the relationship between the echocardiographically determined average flow rate during the early portion of the PV D wave and the average flow rate during the PV S wave (r = 0.76). We conclude that the end-systolic deviation from constant volume for the left heart requires the generation of the LACV during diastole. Because early rapid filling of the left ventricle is the driving force for LACV generation while the left atrium remains passive, it may be more appropriate to consider LACV to be a property of ventricular diastolic rather than atrial function. PMID- 14751860 TI - Myocardial carnitine palmitoyltransferase I expression and long-chain fatty acid oxidation in fetal and newborn lambs. AB - Carnitine palmitoyltransferase I (CPT I) catalyzes the conversion of acyl-CoA to acylcarnitine at the outer mitochondrial membrane and is a key enzyme in the control of long-chain fatty acid (LC-FA) oxidation. Because myocardial LC-FA oxidation increases dramatically after birth, we determined the extent to which CPT I expression contributes to these changes in the perinatal lamb. We measured the steady-state level of transcripts of the CPT1A and CPT1B genes, which encode the liver (L-CPT I) and muscle CPT I (M-CPT I) isoforms, respectively, as well as the amount of these proteins, their total activity, and the amount of carnitine in left ventricular tissue from fetal and newborn lambs. We compared these data with previously obtained myocardial FA oxidation rates in vivo in the same model. The results showed that CPT1B was already expressed before birth and that total CPT I expression transiently increased after birth. The protein level of M-CPT I was high throughout development, whereas that of L-CPT I was only transiently upregulated in the first week after birth. The total CPT I activity in vitro also increased after birth. However, the increase in myocardial FA oxidation measured in vivo (112-fold) by far exceeded the increase in gene expression (2.2-fold), protein amount (1.1-fold), and enzyme activity (1.2-fold) in vitro. In conclusion, these results stress the importance of substrate supply per se in the postnatal increase in myocardial FA oxidation. M-CPT I is expressed throughout perinatal development, making it a primary target for metabolic modulation of myocardial FA oxidation. PMID- 14751862 TI - Respiratory sinus arrhythmia, cardiac vagal control, and daily activity. AB - Ambulatory respiratory sinus arrhythmia (RSA) or high-frequency heart rate (HR) variability is frequently employed as an index of cardiac parasympathetic control and related to risk or severity of cardiovascular disease. However, laboratory studies indicate variations in physical activity and respiratory parameters of rate and tidal volume may confound estimation of vagal activity. Because little is known about these relations outside the laboratory, we examined ambulatory relations among RSA, respiration, physical activity, and HR during waking hours by employing a multichannel monitoring system. Forty healthy young-to-middle aged adults underwent daytime monitoring that included continuous registration of the ECG, respiration (inductance plethysmography), and accelerometry motion activity. Within-individual regression analyses were performed to examine minute-to-minute relations between RSA and respiration, HR, and indexes of physical activity (minute ventilation and motion). HR changes were assumed to be strongly related to within-individual variations of vagal tone. RSA adjusted for respiratory parameters and unadjusted RSA were compared for strength of prediction of other measures. Unadjusted RSA was related to respiratory parameters (R = 0.80) and moderately predicted minute-to-minute HR and activity variances (means = 56%, HR; 48%, minute ventilation; and 37%, motion). Adjusted RSA predicted significantly more HR and activity variance (means = 75%, 76%, and 57%, respectively) with narrower confidence intervals. We conclude that ambulatory RSA magnitude is associated with respiratory variations and physical activity. Adjustment for respiratory parameters substantially improves relations between RSA and significantly vagally mediated HR and physical activity. Concurrent monitoring of respiration and physical activity may enhance HR variability accuracy to predict autonomic control. PMID- 14751861 TI - Reduced release of nitric oxide to shear stress in mesenteric arteries of aged rats. AB - We hypothesized that aging is characterized by a reduced release of nitric oxide (NO) in response to shear stress in resistance vessels. Mesenteric arterioles and arteries of young (6 mo) and aged (24 mo) male Fischer 344 rats were isolated and cannulated. Shear stress (15 dyn/cm(2))-induced dilation was significantly reduced and shear stress (1, 5, 10, and 15 dyn/cm(2))-induced increases in perfusate nitrite were significantly smaller at all shear stress levels in vessels of aged rats. Inhibition of NO synthesis abolished shear stress-induced release of nitrite. Furthermore, shear stress (15 dyn/cm(2))-induced release of nitrate was significantly higher and total nitrite (nitrite plus nitrate) was significantly lower in vessels of aged rats. Tiron or SOD significantly increased nitrite released from vessels of aged rats, but this was still significantly less than that in young rats. Superoxide production was increased and the activity of SOD was decreased in vessels of aged rats. There were no differences in endothelial NO synthase (eNOS) protein and basal activity or in Cu/Zn-SOD and Mn SOD proteins in vessels of the two groups, but extracellular SOD was significantly reduced in vessels of aged rats. Maximal release of NO induced by shear stress plus ACh (10(-5) M) was comparable in the two groups, but phospho eNOS in response to shear stress (15 dyn/cm(2)) was significantly reduced in vessels of aged rats. These data suggest that an increased production of superoxide, a reduced activity of SOD, and an impaired shear stress-induced activation of eNOS are the causes of the decreased shear stress-induced release of NO in vessels of aged rats. PMID- 14751863 TI - Suppression of alternans and conduction blocks despite steep APD restitution: electrotonic, memory, and conduction velocity restitution effects. AB - We examine the utility of the action potential (AP) duration (APD) restitution curve slope in predicting the onset of electrical alternans when electrotonic and memory effects are considered. We develop and use two ionic cell models without memory that have the same restitution curve with slope >1 but different AP shapes and, therefore, different electrotonic effects. We also study a third cell model that incorporates short-term memory of previous cycle lengths, so that it has a family of S1-S2 restitution curves as well as a dynamic restitution curve with slope >1. Our results indicate that both electrotonic and memory effects can suppress alternans, even when the APD restitution curve is steep. In the absence of memory, electrotonic currents related to the shape of the AP, as well as conduction velocity restitution, can affect how alternans develops in tissue and, in some cases, can prevent its induction entirely, even when isolated cells exhibit alternans. When short-term memory is included, alternans may not occur in isolated cells, despite a steep APD restitution curve, and may or may not occur in tissue, depending on conduction velocity restitution. We show for the first time that electrotonic and memory effects can prevent conduction blocks and stabilize reentrant waves in two and three dimensions. Thus we find that the slope of the APD restitution curve alone does not always well predict the onset of alternans and that incorporating electrotonic and memory effects may provide a more useful alternans criterion. PMID- 14751864 TI - Individual differences in respiratory sinus arrhythmia. AB - To investigate the interindividual differences in respiratory sinus arrhythmia (RSA), recordings of ventilation and electrocardiogram were obtained from 12 healthy subjects for five imposed breathing periods (T(TOT)) surrounding each individual's spontaneous breathing period. In addition to the spectral analysis of the R-R interval signal at each breathing period, RSA characteristics were quantified by using a breath-by-breath analysis where a sinusoid was fitted to the changes in instantaneous heart rate in each breath. The amplitude and phase (or delay = phase x T(TOT)) of this sinusoid were taken as the RSA characteristics for each breath. It was found that for each subject the RSA amplitude-T(TOT) relationship was linear, whereas the delay-T(TOT) relationship was parabolic. However, the parameters of these relationships differed between individuals. Linear correlation between the slopes of RSA amplitude versus T(TOT) regression lines and 1) mean breathing period and 2) mean R-R interval during spontaneous breathing were calculated. Only the correlation coefficient with breathing period was significantly different from zero, indicating that the longer the spontaneous breathing period the lesser the increase in RSA amplitude with increasing breathing period. Similarly, only the correlation coefficient between the curvature of the RSA delay-T(TOT) parabola and mean breathing period was significantly different from zero; the longer the spontaneous breathing period the larger the curvature of RSA delay. These results suggest that the changes in RSA characteristics induced by changing the breathing period may be explained partly by the spontaneous breathing period of each individual. Furthermore, a transfer function analysis performed on these data suggested interindividual differences in the autonomic modulation of the heart rate. PMID- 14751866 TI - Thoracic vein ablation terminates chronic atrial fibrillation in dogs. AB - The thoracic vein hypothesis of chronic atrial fibrillation (AF) posits that rapid, repetitive activations from muscle sleeves within thoracic veins underlie the mechanism of sustained AF. If this is so, thoracic vein ablation should terminate sustained AF and prevent its reinduction. Six female mongrel dogs underwent chronic pulmonary vein (PV) pacing at 20 Hz to induce sustained (>48 h) AF. Bipolar electrodes were used to record from the atria and thoracic veins, including the vein of Marshall, four PVs, and the superior vena cava. Radio frequency (RF) application was applied around the PVs and superior vena cava and along the vein of Marshall until electrical activity was eliminated. Computerized mapping (1,792 electrodes, 1 mm resolution) was also performed. Sustained AF was induced in 30.6 +/- 6.5 days, and ablation was done 17.3 +/- 8.5 days afterward. Before ablation, the PVs had shorter activation cycle lengths than the atria, and rapid, repetitive activations were observed in the PVs. All dogs converted to sinus rhythm during (n = 4 dogs) or within 90 min of completion of RF ablation. Rapid atrial pacing afterward induced only nonsustained (<60 s) AF in all dogs. Average AF cycle lengths after reinduction were significantly (P = 0.01) longer (183 +/- 31.5 ms) than baseline (106 +/- 16.2 ms). There were no activation cycle length gradients after RF application. We conclude that thoracic vein ablation converts canine sustained AF into sinus rhythm and prevents the reinduction of sustained AF. These findings suggest that thoracic veins are important in the maintenance of AF in dogs. PMID- 14751865 TI - Platelet-derived growth factor-BB and Ets-1 transcription factor negatively regulate transcription of multiple smooth muscle cell differentiation marker genes. AB - Platelet-derived growth factor (PDGF)-BB, a potent mitogen for mesenchymal cells, also downregulates expression of multiple smooth muscle (SM) cell (SMC)-specific markers. However, there is conflicting evidence whether PDGF-BB represses SMC marker expression at a transcriptional or posttranscriptional level, and little is known regarding the mechanisms responsible for these effects. Results of the present studies provide clear evidence that PDGF-BB treatment strongly repressed SM alpha-actin, SM myosin heavy chain (MHC), and SM22alpha promoters in SMCs. Of major significance for resolving previous controversies in the field, we found PDGF-BB-induced repression of SMC marker gene promoters in subconfluent, but not postconfluent, cultures. Treatment of postconfluent SMCs with a tyrosine phosphatase inhibitor restored PDGF-BB-induced repression, whereas treatment of subconfluent SMCs with a tyrosine kinase blocker abolished PDGF-BB-induced repression, suggesting that a tyrosine phosphorylation event mediates cell density-dependent effects. On the basis of previous observations that Ets-1 transcription factor is upregulated within phenotypically modulated neointimal SMCs, we tested whether Ets-1 would repress SMC marker expression. Consistent with this hypothesis, results of cotransfection experiments indicated that Ets-1 overexpression reduced transcriptional activity of SMC marker promoter constructs in SMCs, whereas it increased activity of SM alpha-actin promoter in endothelial cells. PDGF-BB treatment increased expression of Ets-1 in cultured SMCs, and SM alpha-actin mRNA expression was reduced in multiple independent clones of SMCs stably transfected with an Ets-1-overexpressing construct. Taken together, results of these experiments provide novel insights regarding possible mechanisms whereby PDGF-BB and Ets-1 may contribute to SMC phenotypic switching associated with vascular injury. PMID- 14751867 TI - Efficacy of preconditioning should be gauged by reduction of infarction. PMID- 14751868 TI - Identification of interstitial cells of Cajal in corporal tissues of the guinea pig penis. AB - This study shows for the first time the presence of interstitial cells of Cajal (ICC) and their possible role in the initiation of spontaneous excitation in the corporal tissue of the guinea-pig penis. ICC, which were identified by their c kit immunoreactivity, were abundantly distributed in the corporal smooth muscle meshwork. Spontaneous increases in the intracellular calcium concentration ([Ca(2+)](i); calcium transients) were visualized in preparations loaded with the fluorescent dye fura-2. Ca transients originated from the boundary of muscle bundles and then spread throughout the meshwork (Ca waves). Ca waves were strongly suppressed by either CPA (10 microm), ryanodine (50 microm) or 2-APB (10 microm), and their synchronicity was disrupted by 18beta-GA (30 microm). These results suggest that ICC in the corporal tissue may have a role as pacemakers to drive the bulk of smooth muscles, and that intracellular Ca(2+) stores and gap junctions are critical for the generation of spontaneous excitation. PMID- 14751869 TI - A1 and A2A adenosine receptor modulation of alpha 1-adrenoceptor-mediated contractility in human cultured prostatic stromal cells. AB - 1. This study investigated the possibility that adenosine receptors modulate the alpha(1)-adrenoceptor-mediated contractility of human cultured prostatic stromal cells (HCPSC). 2. The nonselective adenosine receptor agonist, 5'-N ethylcarboxamido-adenosine (NECA; 10 nm-10 microm), and the A(1) adenosine receptor selective agonist, cyclopentyladenosine (CPA; 10 nm-10 microm), elicited significant contractions in HCPSC, with maximum contractile responses of 18+/-3% and 17+/-2% reduction in initial cell length, respectively. 3. In the presence of a threshold concentration of phenylephrine (PE) (100 nm), CPA (1 nm-10 microm) caused contractions, with an EC(50) of 124+/-12 nm and maximum contractile response of 37+/-4%. The A(1) adenosine receptor-selective antagonist 8 cyclopentyl-1,3-dipropylxanthine (DPCPX 100 nm) blocked this effect. In the presence of DPCPX (100 nm), NECA (1 nm-10 microm) inhibited contractions elicited by a submaximal concentration of PE (10 microm), with an IC(50) of 48+/-2 nm. The A(2A) adenosine receptor-selective antagonist 4-(2-[7-amino-2 [furyl][1,2,4]triazolo[2,3-alpha][1,3,5,]triazin-5-yl amino]ethyl)phenol (Zm241385 100 nm) blocked this effect. 4. In BCECF-AM (10 microm)-loaded cells, both CPA (100 pM-1 microm) and NECA (100 pm-10 microm) elicited concentration dependent decreases in intracellular pH (pH(i)), with EC(50) values of 3.1+/-0.3 and 6.0+/-0.3 nm, respectively. The response to NECA was blocked by Zm241385 (100 nm; apparent pK(B) of 9.4+/-0.4), but not by DPCPX (100 nm). The maximum response to CPA was blocked by DPCPX (100 nm), and unaffected by Zm241385 (100 nm). 5. NECA (10 nm-10 microm) alone did not increase [(3)H]-cAMP in HCPSC. In the presence of DPCPX (100 nm), NECA (10 nm-10 microm) caused a concentration dependent increase in [(3)H]-cAMP, with an EC(50) of 1.2+/-0.1 microm. This response was inhibited by Zm241385 (100 nm). CPA (10 nm-10 microm) had no effect on cAMP, in the presence or absence of forskolin (1 microm). 6. These findings are consistent with a role for adenosine receptors in the modulation of adrenoceptor-mediated contractility in human prostate-derived cells. PMID- 14751872 TI - Pandemic risks from bird flu. PMID- 14751870 TI - Characterization of ERK1/2 signalling pathways induced by adenosine receptor subtypes in newborn rat cardiomyocytes. AB - 1. Adenosine A(1), A(2A), and A(3) receptors (ARs) and extracellular signal regulated kinase 1/2 (ERK1/2) play a major role in myocardium protection from ischaemic injury. In this study, we have characterized the adenosine receptor subtypes involved in ERK1/2 activation in newborn rat cardiomyocytes. 2. Adenosine (nonselective agonist), CPA (A(1)), CGS 21680 (A(2A)) or Cl-IB-MECA (A(3)), all increased ERK1/2 phosphorylation in a time- and dose-dependent manner. The combined maximal response of the selective agonists was similar to adenosine alone. Theophylline (nonselective antagonist) inhibited completely adenosine-mediated ERK1/2 activation, whereas a partial inhibition was obtained with DPCPX (A(1)), ZM 241385 (A(2A)), and MRS 1220 (A(3)). 3. PD 98059 (MEK1; ERK kinase inhibitor) abolished all agonist-mediated ERK1/2 phosphorylation. Pertussis toxin (PTX, G(i/o) blocker) inhibited completely CPA- and partially adenosine- and Cl-IB-MECA-induced ERK1/2 activation. Genistein (tyrosine kinase inhibitor) and Ro 318220 (protein kinase C, PKC inhibitor) partially reduced adenosine, CPA and Cl-IB-MECA responses, without any effect on CGS 21680-induced ERK1/2 phosphorylation. H89 (protein kinase A, PKA inhibitor) abolished completely CGS 21680 and partially adenosine and Cl-IB-MECA responses, without any effect on CPA response. 4. Cl-IB-MECA-mediated increases in cAMP accumulation suggest that A(3)AR-induced ERK1/2 phosphorylation involves adenylyl cyclase activation via phospholipase C (PLC) and PKC stimulation. 5. In summary, we have shown that ERK1/2 activation by adenosine in cardiomyocytes results from an additive stimulation of A(1), A(2A), and A(3)ARs, which involves G(i/o) proteins, PKC, and tyrosine kinase for A(1) and A(3)ARs, and Gs and PKA for A(2A)ARs. Moreover, the A(3)AR response also involves a cAMP/PKA pathway via PKC activation. PMID- 14751871 TI - Radical surgery for mesothelioma. PMID- 14751873 TI - Exchanging health lessons globally. PMID- 14751874 TI - Implementing the European clinical trials directive. PMID- 14751875 TI - Access to antiretroviral treatment in Africa. PMID- 14751876 TI - Death toll mounts in avian flu outbreak. PMID- 14751877 TI - Judge may report drug company executive to attorney general. PMID- 14751878 TI - Shortage of sperm donors predicted when anonymity goes. PMID- 14751880 TI - United States wins more time to lobby against WHO diet plan. PMID- 14751881 TI - Psychological therapies recommended for eating disorders. PMID- 14751883 TI - Payments announced for patients infected with hepatitis C. PMID- 14751884 TI - Kidney trade arrest exposes loopholes in India's transplant laws. PMID- 14751886 TI - Doctors suspended for removing wrong kidney. PMID- 14751888 TI - Controversy surrounds proposed Italian alternative to female genital mutilation. PMID- 14751890 TI - Australian drug pricing scheme under pressure in free trade talks. PMID- 14751892 TI - Report calls for strategies to reduce medication errors. PMID- 14751893 TI - Parents discover their stillborn babies are alive four months after birth. PMID- 14751896 TI - Changing practice. PMID- 14751898 TI - Clinical risk management in obstetrics: eclampsia drills. AB - PROBLEM: Infrequent presentation of patients with eclampsia, leading to staff inexperienced in the condition and untested emergency systems. DESIGN: "Fire drill" programme using on-site simulation of patients with eclampsia. SETTING: Tertiary referral obstetric unit. KEY MEASURES FOR IMPROVEMENT: Successful implementation of measures to optimise management of eclampsia. STRATEGIES FOR CHANGE: Rapid activation of emergency team after one call, development and dissemination of evidence based protocol for eclampsia, strategically placed "eclampsia boxes," individual staff feedback and education. EFFECTS OF CHANGE: Efficient and appropriate management of subsequent simulated patients. LESSONS LEARNT: On-site simulation can identify and correct potential deficiencies in the care of patients with eclampsia. PMID- 14751897 TI - Indirect comparisons: a novel approach to assessing the effect of anti-HIV drugs. PMID- 14751899 TI - How to investigate and manage the child who is slow to speak. PMID- 14751901 TI - Why people smoke. PMID- 14751902 TI - Antiretroviral therapy in Africa. PMID- 14751903 TI - Evaluating the health effects of social interventions. PMID- 14751904 TI - Save European research campaign. PMID- 14751905 TI - Quality of randomised controlled trials: caution is important. PMID- 14751906 TI - Quality of randomised controlled trials: quality of research may be worse than it appears. PMID- 14751907 TI - Quality of randomised controlled trials: quality of trial methods is not good in all disciplines. PMID- 14751908 TI - WHO's world health report 2003: actions speak louder than words. PMID- 14751909 TI - Stress and exacerbations in multiple sclerosis: whether stress triggers relapses remains a conundrum. PMID- 14751910 TI - Special issue on South Asia: health economics is neglected in this region. PMID- 14751911 TI - Special issue on South Asia: focus will be an eye opener. PMID- 14751912 TI - Colchicine in acute gout: optimal dose of colchicine is still elusive. PMID- 14751913 TI - Colchicine in acute gout: low dose colchicine was started after usual dose. PMID- 14751914 TI - WHO's world health report 2003: time to ease the mental health burden. PMID- 14751915 TI - Polypill debate continues: concept is a fascinating thought experiment. PMID- 14751916 TI - Polypill debate continues: people will always be sceptical. PMID- 14751917 TI - Polypill debate continues: similar combination of drugs has worked in natural experiment. PMID- 14751918 TI - Retrospective cohort study of false alarm rates associated with a series of heart operations: the case for hospital mortality monitoring groups. AB - OBJECTIVE: To examine the efficacy of different methods of detecting a high death rate and determining whether an increase in deaths after heart transplantation could be explained by chance. DESIGN: Retrospective analysis of deaths after heart transplantation. Seven methods were used: mortality above national average, mortality excessively above national average, test of moving average mortality, test of number of consecutive deaths, sequential probability ratio test (SPRT), cusum graph with v-mask, and CRAM chart. The national average mortality was not available and a rate of 15% was used instead as the benchmark. SETTING: Regional cardiothoracic unit. PARTICIPANTS: All 371 patients who received a heart transplant in the programme, 1986-2000. MAIN OUTCOME MEASURES: 30 day survival after transplantation. RESULTS: All methods provided evidence that the 30 day mortality had been high at some stage. The probability that the finding was a false positive depended on which test was used. At the end of the series the average mortality, sequential probability ratio, and cusum tests indicated a level of deaths higher than the benchmark while the remaining four tests yielded negative results. CONCLUSIONS: If the decision to test for outlying mortality is made retrospectively, in the light of the data, it is not possible to determine the false positive rate. Prospective on-site mortality monitoring with the CRAM chart is recommended as this method can quantify the death rate and identify periods when an audit of cases is indicated, even when data from other institutions are not available. A hospital mortality monitoring group can routinely monitor all deaths in the hospital, by specialty, using hospital episode statistics (HES) data and appropriate statistical methods. PMID- 14751919 TI - Patients' preferences for the management of non-metastatic prostate cancer: discrete choice experiment. AB - OBJECTIVE: To establish which attributes of conservative treatments for prostate cancer are most important to men. DESIGN: Discrete choice experiment. SETTING: Two London hospitals. PARTICIPANTS: 129 men with non-metastatic prostate cancer, mean age 70 years; 69 of 118 (58%) with T stage 1 or 2 cancer at diagnosis. MAIN OUTCOME MEASURES: Men's preferences for, and trade-offs between, the attributes of diarrhoea, hot flushes, ability to maintain an erection, breast swelling or tenderness, physical energy, sex drive, life expectancy, and out of pocket expenses. RESULTS: The men's responses to changes in attributes were all statistically significant. When asked to assume a starting life expectancy of five years, the men were willing to make trade-offs between life expectancy and side effects. On average, they were most willing to give up life expectancy to avoid limitations in physical energy (mean three months) and least willing to trade life expectancy to avoid hot flushes (mean 0.6 months to move from a moderate to mild level or from mild to none). CONCLUSIONS: Men with prostate cancer are willing to participate in a relatively complex exercise that weighs up the advantages and disadvantages of various conservative treatments for their condition. They were willing to trade off some life expectancy to be relieved of the burden of troublesome side effects such as limitations in physical energy. PMID- 14751920 TI - Treatments of homosexuality in Britain since the 1950s--an oral history: the experience of patients. AB - OBJECTIVES: To investigate the circumstances since the 1950s in which people who were attracted to members of the same sex received treatments to change their sexual orientation, the referral pathway and the process of therapy, and its aftermath. DESIGN: A nationwide study based on qualitative interviews. PARTICIPANTS: 29 people who had received treatments to change their sexual orientation in the United Kingdom and two relatives of former patients. RESULTS: Most participants had been distressed by their attraction to their own sex and people in whom they confided thought they needed treatment. Although some participants chose to undergo treatments instead of imprisonment or were encouraged through some form of medical coercion, most were responding to complex personal and social pressures that discouraged any expression of their sexuality. While many participants found happiness in same sex relationships after their treatment, most were left feeling emotionally distressed to some degree. CONCLUSION: The definition of same sex attraction as an illness and the development of treatments to eradicate such attraction have had a negative long term impact on individuals. PMID- 14751921 TI - Treatments of homosexuality in Britain since the 1950s--an oral history: the experience of professionals. AB - OBJECTIVE: To investigate the experiences of professionals who administered and evaluated treatments for homosexuality in Britain since the 1950s. DESIGN: A nationwide study based on qualitative interviews. PARTICIPANTS: 30 health professionals who developed and practised treatments for homosexuality. RESULTS: A range of treatments were developed to make homosexuals into heterosexuals, the most common of which were behavioural interventions. Treatments were based on little evidence of effectiveness and were open to the criticism that legal or social pressures coerced patients. Treatments did not become mainstream within British mental health services. With hindsight, professionals realised that they had not appreciated the influence of social context on sexual behaviour. Most now regarded same sex attraction as compatible with psychological health, although a small minority considered that the option to try to become heterosexual should still be available to patients who desire it. CONCLUSIONS: Social and political assumptions sometimes lie at the heart of what we regard as mental pathology and serve as a warning for future practice. PMID- 14751922 TI - Effect of hepcidin on intestinal iron absorption in mice. AB - The effect of the putative iron regulatory peptide hepcidin on iron absorption was investigated in mice. Hepcidin peptide was synthesized and injected into mice for up to 3 days, and in vivo iron absorption was measured with tied-off segments of duodenum. Liver hepcidin expression was measured by reverse transcriptase polymerase chain reaction. Hepcidin significantly reduced mucosal iron uptake and transfer to the carcass at doses of at least 10 microg/mouse per day, the reduction in transfer to the carcass being proportional to the reduction in iron uptake. Synthetic hepcidin injections down-regulated endogenous liver hepcidin expression excluding the possibility that synthetic hepcidin was functioning by a secondary induction of endogenous hepcidin. The effect of hepcidin was significant at least 24 hours after injection of hepcidin. Liver iron stores and hemoglobin levels were unaffected by hepcidin injection. Similar effects of hepcidin on iron absorption were seen in iron-deficient and Hfe knockout mice. Hepcidin inhibited the uptake step of duodenal iron absorption but did not affect the proportion of iron transferred to the circulation. The effect was independent of iron status of mice and did not require Hfe gene product. The data support a key role for hepcidin in the regulation of intestinal iron uptake. PMID- 14751923 TI - Mechanisms and implications of phosphoinositide 3-kinase delta in promoting neutrophil trafficking into inflamed tissue. AB - The phosphoinositide 3-kinase (PI3K) catalytic subunit p110 delta is expressed in neutrophils and is thought to play a role in their accumulation at sites of inflammation by contributing to chemoattractant-directed migration. We report here that p110 delta is present in endothelial cells and participates in neutrophil trafficking by modulating the proadhesive state of these cells in response to tumor necrosis factor alpha (TNF alpha). Specifically, administration of the selective inhibitor of PI3K delta, IC87114, to animals reduced neutrophil tethering to and increased rolling velocities on cytokine-activated microvessels in a manner similar to that observed in mice deficient in p110 delta. These results were confirmed in vitro as inhibition of this isoform in endothelium, but not neutrophils, diminished cell attachment in flow. A role for PI3K delta in TNF alpha-induced signaling is demonstrated by a reduction in Akt-phosphorylation and phosphatidylinositol-dependent kinase 1 (PDK1) enzyme activity upon treatment of this cell type with IC87114. p110 delta expressed in neutrophils also contributes to trafficking as demonstrated by the impaired movement of these cells across inflamed venules in animals in which this catalytic subunit was blocked or genetically deleted, results corroborated in transwell migration assays. Thus, PI3K delta may be a reasonable therapeutic target in specific inflammatory conditions as blockade of its activity reduces neutrophil influx into tissues by diminishing their attachment to and migration across vascular endothelium. PMID- 14751924 TI - Allogeneic bone marrow transplantation for children with acute myelocytic leukemia in first remission demonstrates a role for graft versus leukemia in the maintenance of disease-free survival. AB - In Children's Cancer Group (CCG) study 2891, patients who were recently diagnosed with acute myelocytic leukemia (AML) were assigned randomly to standard- or intensive-timing induction chemotherapy. Patients in first complete remission (CR1) and who had a human leukocyte antigen (HLA)-identical, related donor or a donor disparate at a single class I or II locus were nonrandomly assigned to receive a bone marrow transplant (BMT) by using oral busulfan (16 mg/kg) and cyclophosphamide (200 mg/kg). Methotrexate only was given for graft-versus-host disease (GVHD) prophylaxis. One hundred fifty patients received transplants. Grade 3 or 4 acute GVHD occurred in 9% of patients. Patients younger than 10 years had a lower incidence of grade 3 or 4 GVHD (4.6%) compared with patients 10 years or older (17.4%) (P =.044). Disease-free survival (DFS) at 6 years was 67% and 42% for recipients of intensive- and standard-timing induction therapies, respectively. Multivariate analysis showed that receiving intensive-timing induction therapy (P =.027) and having no hepatomegaly at diagnosis (P =.009) was associated with favorable DFS, and grades 3 and 4 acute GVHD were associated with inferior DFS. Multivariate analysis showed that grades 1 or 2 GVHD (P =.008) and no hepatomegaly at diagnosis (P =.014) were associated with improved relapse-free survival (RFS). Our results show that children older than 10 years are at higher risk for developing severe GVHD; acute GVHD is associated with favorable RFS. PMID- 14751925 TI - Stroke and conversion to high risk in children screened with transcranial Doppler ultrasound during the STOP study. AB - The Stroke Prevention Trial in Sickle Cell Anemia (STOP) was a randomized multicenter controlled trial comparing prophylactic blood transfusion with standard care in sickle cell anemia (SCA) children aged 2 to 16 years selected for high stroke risk by transcranial Doppler (TCD). More than 2000 children were screened with TCD to identify the 130 high-risk children who entered the randomized trial. A total of 5613 TCD studies from 2324 children were evaluated. We also collected information on stroke. We describe the changes in TCD with repeated testing and report the outcome without transfusion in the STOP screened cohort. Risk of stroke was higher with abnormal TCD than with normal or conditional TCD (P <.001) or inadequate TCD (P =.002), and risk with conditional TCD was higher than with normal TCD (P <.001). Repeated TCD in 1215 children showed that the condition of 9.4% of children became abnormal during observation. Younger patients and those with higher initial flow velocities were most likely to convert to abnormal TCDs. Screening in STOP confirmed the predictive value of TCD for stroke. Substantial differences in the probability of conversion to abnormal TCD were observed, with younger children and those with higher velocity more likely to have an abnormal TCD with rescreening. PMID- 14751926 TI - Hephaestin is a ferroxidase that maintains partial activity in sex-linked anemia mice. AB - Hephaestin (Hp) plays an important role in intestinal iron absorption and is predicted to be a ferroxidase based on significant sequence identity to the serum multicopper ferroxidase ceruloplasmin. Here, we demonstrate that Hp has both amine oxidase and ferroxidase activity in cultured cells and primary intestinal enterocytes with the use of both gel and solution assays. The specificity of the activity is shown by immunoblotting, immunoprecipitation, and immunodepletion experiments. Surprisingly, the truncated hephaestin expressed in sex-linked anemia (sla) mice still has measurable, but decreased, oxidase activity. Molecular modeling of the truncated hephaestin suggests retention of a minimum catalytic core required for enzymatic activity. We suggest that hephaestin, by way of its ferroxidase activity, facilitates iron export from intestinal enterocytes, most likely in cooperation with the basolateral iron transporter, Ireg1. PMID- 14751927 TI - Aberrant quantity and localization of Aurora-B/AIM-1 and survivin during megakaryocyte polyploidization and the consequences of Aurora-B/AIM-1-deregulated expression. AB - Megakaryocytes skip late anaphase and cytokinesis during endomitosis. We found normal expression and localization of a fundamental regulator of mitosis, Aurora B/AIM-1, during prophase in polyploidizing mouse bone marrow megakaryocytes. At late anaphase, however, Aurora-B/AIM-1 is absent or mislocalized. Megakaryocytes treated with a proteasome inhibitor display Aurora-B/AIM-1 properly expressed and localized to the midzone, suggesting that protein degradation contributes to this atypical appearance. In contrast, survivin, an Aurora-B/AIM-1 coregulator of mitosis, is not detected at any stage of the endomitotic cell cycle, and in most megakaryocytes proteasome inhibition does not rescue this phenotype. To further explore the importance of reduced Aurora-B/AIM-1 for polyploidization, it was overexpressed in megakaryocytes of transgenic mice. The phenotype includes increased transgenic mRNA, but not protein, in polyploidy megakaryocytes, further suggesting that Aurora-B/AIM-1 is regulated at the protein level. Aurora-B/AIM-1 protein is, however, elevated in diploid transgenic megakaryocytes. Transgenic mice also exhibit enhanced numbers of megakaryocytes with increased proliferative potential, and some mice exhibit mild decreases in ploidy level. Hence, the molecular programming involved in endomitosis is characterized by the mislocalization or absence of at least 2 critical mitotic regulators, Aurora B/AIM-1 and survivin. Future studies will examine the impact of survivin restoration on mouse megakaryocyte polyploidization. PMID- 14751928 TI - The small oligomerization domain of gephyrin converts MLL to an oncogene. AB - The MLL (mixed lineage leukemia) gene forms chimeric fusions with a diverse set of partner genes as a consequence of chromosome translocations in leukemia. In several fusion partners, a transcriptional activation domain appears to be essential for conferring leukemogenic capacity on MLL protein. Other fusion partners, however, lack such domains. Here we show that gephyrin (GPHN), a neuronal receptor assembly protein and rare fusion partner of MLL in leukemia, has the capacity as an MLL-GPHN chimera to transform hematopoietic progenitors, despite lack of transcriptional activity. A small 15-amino acid tubulin-binding domain of GPHN is necessary and sufficient for this activity in vitro and in vivo. This domain also confers oligomerization capacity on MLL protein, suggesting that such activity may contribute critically to leukemogenesis. The transduction of MLL-GPHN into hematopoietic progenitor cells caused myeloid and lymphoid lineage leukemias in mice, suggesting that MLL-GPHN can target multipotent progenitor cells. Our results, and other recent data, provide a mechanism for oncogenic conversion of MLL by fusion partners encoding cytoplasmic proteins. PMID- 14751929 TI - Deregulated expression in Ph+ human leukemias of AHI-1, a gene activated by insertional mutagenesis in mouse models of leukemia. AB - Ahi-1/AHI-1 (Abelson helper integration site-1) encodes a family of protein isoforms containing one Src homology 3 (SH3) domain and multiple tryptophan aspartic acid 40 (WD40)-repeat domains. The function of these proteins is unknown, but involvement in leukemogenesis has been suggested by the high frequency of Ahi-1 mutations seen in certain virus-induced murine leukemias. Here we show that in both mice and humans, Ahi-1/AHI-1 expression is highest in the most primitive hematopoietic cells with specific patterns of down-regulation in different lineages. Cells from patients with chronic myeloid leukemia (CML; n = 28) show elevated AHI-1 transcripts in all disease phases and, in chronic phase, in the leukemic cells at all stages of differentiation, including quiescent (G(0)) CD34(+) cells as well as terminally differentiating cells. In the most primitive lin(-)CD34(+)CD38(-) CML cells, transcripts for the 2 shorter isoforms of AHI-1 are also increased. Although 15 of 16 human lymphoid and myeloid leukemic cell lines showed aberrant control of AHI-1 expression, this was not seen in blasts obtained directly from patients with acute Philadelphia chromosome negative (Ph(-)) leukemia (n = 15). Taken together, our results suggest that down regulation of AHI-1 expression is an important conserved step in primitive normal hematopoietic cell differentiation and that perturbations in AHI-1 expression may contribute to the development of specific types of human leukemia. PMID- 14751930 TI - A novel plasma proteinase potentiates alpha2-antiplasmin inhibition of fibrin digestion. AB - Human alpha2-antiplasmin (alpha2AP), also known as alpha2-plasmin inhibitor, is the major inhibitor of the proteolytic enzyme plasmin that digests fibrin. There are 2 N-terminal forms of alpha2AP that circulate in human plasma: a 464-residue protein with Met as the N-terminus, Met-alpha2AP, and a 452-residue version with Asn as the N-terminus, Asn-alpha2AP. We have discovered and purified a proteinase from human plasma that cleaves the Pro12-Asn13 bond of Met-alpha2AP to yield Asn alpha2AP and have named it antiplasmin-cleaving enzyme (APCE). APCE is similar in primary structure and catalytic properties to membrane-bound fibroblast activation protein/seprase for which a physiologic substrate has not been clearly defined. We found that Asn-alpha2AP becomes cross-linked to fibrin by activated factor XIII approximately 13 times faster than native Met-alpha2AP during clot formation and that clot lysis rates are slowed in direct proportion to the ratio of Asn-alpha2AP to Met-alpha2AP in human plasma. We conclude that APCE cleaves Met-alpha2AP to the derivative Asn-alpha2AP, which is more efficiently incorporated into fibrin and consequently makes it strikingly resistant to plasmin digestion. APCE may represent a new target for pharmacologic inhibition, since less generation and incorporation of Asn-alpha2AP could result in a more rapid removal of fibrin by plasmin during atherogenesis, thrombosis, and inflammatory states. PMID- 14751931 TI - Prompt versus preemptive intervention for EBV lymphoproliferative disease. AB - Posttransplantation lymphoproliferative disorders (PTLDs) caused by uncontrolled expansion of Epstein-Barr virus (EBV)-infected B cells after hematopoietic stem cell transplantation (HSCT) can be predicted by an increase in EBV DNA in peripheral blood mononuclear cells. We used real-time quantitative polymerase chain reaction (RQ-PCR) analysis to determine whether frequent monitoring of EBV DNA to allow preemptive treatment is truly of value in patients after HSCT. More than 1300 samples from 85 recipients were analyzed. No patient with consistently low EBV DNA levels developed PTLD. Nine patients had a single episode with a high EBV load (more than 4000 EBV copies/microg peripheral blood mononuclear cell [PBMC] DNA), and 16 patients had high EBV loads detected on 2 or more occasions. Only 8 of these developed symptoms consistent with PTLD, and all were promptly and successfully treated with EBV-specific cytotoxic T cells or CD20 monoclonal antibody. Hence, quantitative measurement of EBV DNA may best be used to enable the prompt rather than the preemptive treatment of PTLD. PMID- 14751933 TI - Should computed tomography of the chest be recommended in the medical certification of professional divers? PMID- 14751934 TI - No pain, no gain? Thoughts on the Caerphilly study. PMID- 14751935 TI - The role of exercise prescription in chronic disease. PMID- 14751937 TI - Genedicine. PMID- 14751936 TI - Eccentric training in patients with chronic Achilles tendinosis: normalised tendon structure and decreased thickness at follow up. AB - OBJECTIVE: To prospectively investigate tendon thickness and tendon structure by ultrasonography in patients treated with eccentric calf muscle training for painful chronic Achilles tendinosis located at the 2-6 cm level in the tendon. METHODS: The patients were examined with grey scale ultrasonography before and 3.8 years (mean) after the 12 week eccentric training regimen. At follow up, a questionnaire assessed present activity level and satisfaction with treatment. RESULTS: Twenty six tendons in twenty five patients (19 men and six women) with a mean age of 50 years were followed for a mean of 3.8 years (range 1.6-7.75). All patients had a long duration of painful symptoms (mean 17.1 months) from chronic Achilles tendinosis before treatment. At follow up, 22 of 25 patients were satisfied with treatment and active in Achilles tendon loading activities at the desired level. Ultrasonography showed that tendon thickness (at the widest part) had decreased significantly (p<0.005) after treatment (7.6 (2.3) v 8.8 (3) mm; mean (SD)). In untreated normal tendons, there was no significant difference in thickness after treatment (5.3 (1.3) mm before and 5.9 (0.8) mm after). All tendons with tendinosis had structural abnormalities (hypoechoic areas and irregular structure) before the start of treatment. After treatment, the structure was normal in 19 of the 26 tendons. Six of the seven patients with remaining structural abnormalities experienced pain in the tendon during loading. CONCLUSIONS: Ultrasonographic follow up of patients with mid-portion painful chronic Achilles tendinosis treated with eccentric calf muscle training showed a localised decrease in tendon thickness and a normalised tendon structure in most patients. Remaining structural tendon abnormalities seemed to be associated with residual pain in the tendon. PMID- 14751938 TI - Prevalence of Staphylococcus aureus carriage by young Malaysian footballers during indoor training. AB - BACKGROUND: Research has shown that athletes are carriers of Staphylococcus aureus during physical activity. OBJECTIVE: To estimate the mean total plate count of S aureus carried by footballers before and after training at an indoor venue. METHODS: Forty Malay and 20 Indian students volunteered to participate. There was also a control group consisting of 40 Malay and 20 Indian students who were not active. The experimental group were active footballers who had played at school or club level. The subjects were healthy and free of skin infection. The experiment was divided into three sessions, with 20 subjects present at each. At each session, the subjects trained for one hour. Swabs were taken from the skin, nose, and ear before and after training. For the control group, swabs were taken only once from the skin, nose, and ear. The swabs were subjected to biochemical tests and then streaked and cultured aerobically in Baird Parker agar plates for 24 hours at 37 degrees C. Black colonies with a clear zone were presumed to be S aureus, and the mean total plate count of the colonies was estimated. Gram staining, catalase, coagulase slide, coagulase tube, acetoin production, o nitrophenyl beta-D-galactopyranoside (ONPG), and mannitol fermentation tests were used to confirm the colonies as S aureus. A haemolysin test was conducted with human blood to confirm haemolytic activity. RESULTS: All subjects in the experimental group were carrying S aureus both before and after training. The estimated mean total counts of colonies from the skin, ear, and nose for the Malays before training were 33, 71, and 312 respectively. Counts after training were 21, 44, and 452 respectively. The results for the Indians were 72, 80, and 309 respectively before training and 55, 200, and 466 respectively after training. The positive results for Gram staining, catalase, coagulase slide, coagulase tube, acetoin production, ONPG, and mannitol fermentation tests were 100%, 96%, 95%, 95%, 93%, 93%, and 90% respectively. All subjects in the control group were also carrying S aureus. CONCLUSIONS: All of the players were carriers of S aureus during training. The decrease in total count from the skin for both races may be due to lysozyme activity lysing the bacterial cells. Contamination of the environment with these bacteria may have increased the estimated total plate count in the nose. The experimental group face a higher risk of infection because of lower immunity during training and higher rate of injuries compared with the control group. PMID- 14751939 TI - 99mTc-MDP bone SPECT in evaluation of the knee in asymptomatic soccer players. AB - OBJECTIVE: To evaluate stress fractures in leg (particularly around the knee, tibia, and femur) and knee pathology in active asymptomatic (no symptoms in the preceding month) soccer players. METHOD: The study included 42 asymptomatic soccer players (21 women, 21 men; age range 19-31 years). Players from seven teams in the major female professional and amateur male soccer leagues were examined by technetium-99m-methylene diphosphonate ((99m)Tc-MDP) bone scintigraphy during the soccer season. Four hours after intravenous injection of 20 mCi (99m)Tc-MDP, standard imaging included anterior planar spot images of the legs, lateral images of the knee, and single photon emission computed tomography (SPECT). RESULTS: Although the players were asymptomatic, increased tracer uptake, indicating stress fracture, was found in 28 (66%). Most of the stress fractures were in the tibia (62%) and femur (5%). In the 42 subjects (84 legs), 35 sites (42%) showed rupture of the posterior horn of the lateral meniscus and bone bruising of the tibial plateau, 16 sites (19%) showed rupture of the anterior horn of the medial meniscus, 11 sites (13%) showed bone bruising of the lateral femoral condyle, eight sites (10%) showed bone bruising of the medial femoral condyle, and there was avulsion injury to the infrapatellar tendon insertion in the anterior tibia in 34 sites (40%). There were 11 anterior cruciate ligament injuries. CONCLUSION: Bone SPECT is very accurate, easy to perform, cost effective, may give valuable information before magnetic resonance imaging studies in the detection of meniscal tears, and may be used successfully when magnetic resonance imaging is unavailable. PMID- 14751940 TI - The Victorian Active Script Programme: promising signs for general practitioners, population health, and the promotion of physical activity. AB - BACKGROUND: The Active Script Programme (ASP) aimed to increase the number of general practitioners (GPs) in Victoria, Australia who deliver appropriate, consistent, and effective advice on physical activity to patients. To maximise GP participation, a capacity building strategy within Divisions of General Practice (DGPs) was used. The objectives of the programme were to (a) train and support GPs in advising sedentary patients, and (b) develop tools and resources to assist GPs. OBJECTIVE: To evaluate the effectiveness of the ASP. METHODS: A systems approach was used to promote capacity in Victorian general practice. Economic analyses were incorporated into the programme's evaluation. Participants were selected DGPs and their GP members. The programme worked with DGPs to train GPs and provide relevant resources. The main outcome measures were (a) changes in GP knowledge and behaviour and (b) cost effectiveness, based on modelled estimates of numbers of patients advised and adopting physical activity and gaining the associated health benefits. RESULTS: GP awareness and provision of physical activity advice increased. Although the programme's reach was modest, based on actual GP involvement, the cost effectiveness figures (138 Australian dollars per patient to become sufficiently active to gain health benefits, and 3647 Australian dollars per disability adjusted life year saved) are persuasive. CONCLUSIONS: The ASP increased DGPs' capacity to support GPs to promote physical activity. There is a strong economic argument for governments to invest in such programmes. However, caution is warranted about the maintenance of patients' activity levels. Programme refinement to encourage GPs to use community supports more effectively will guide future development. Further research on long term patient adherence through a multisectorial approach is warranted. PMID- 14751941 TI - Arthrogenic muscle response to a simulated ankle joint effusion. AB - BACKGROUND: Arthrogenic muscle inhibition (AMI) is a continuing reflex reaction of the musculature surrounding a joint after distension or damage to the structures of that joint. This phenomenon has been well documented after knee joint injury and has been generalised to occur at other joints of the human body, yet minimal research has been conducted in this regard. The response of the muscles crossing the ankle/foot complex after ankle injury and effusion is not well understood. AMI may occur after an ankle sprain contributing to residual dysfunction. OBJECTIVE: To determine if AMI is present in the soleus, peroneus longus, and tibialis anterior musculature after a simulated ankle joint effusion. METHODS: Eight neurologically sound volunteers (mean (SD) age 23 (4) years, height 171 (6) cm, mass 73 (10) kg) participated. Maximum H-reflex and maximum M wave measurements were collected using surface electromyography after delivery of a percutaneous stimulus to the sciatic nerve before its bifurcation into the common peroneal and posterior tibial nerves. RESULTS: The H-reflex and M-wave measurements in all muscles increased (p< or =0.05) after the simulated ankle joint effusion. CONCLUSIONS: Simulated ankle joint effusion results in facilitation of the soleus, peroneus longus, and tibialis anterior motoneurone pools. This may occur to stabilise the foot/ankle complex in order to maintain posture and/or locomotion. PMID- 14751942 TI - Effect of contraction frequency on energy expenditure and substrate utilisation during upper and lower body exercise. AB - OBJECTIVE: To examine the effect of contraction frequency on energy expenditure and substrate utilisation during upper (UE) and lower (LE) body exercise. METHODS: Twenty four college students were recruited: 12 were tested on an arm ergometer, and the other 12 were tested on a leg ergometer. Each subject underwent three experimental trials on three separate days, and the three trials were presented in a randomised order. Each trial consisted of 10 minutes of arm cranking or leg cycling at 40, 60, or 80 rev/min, with power output being kept constant at 50 W. Steady state oxygen uptake (VO(2)) and respiratory exchange ratio (RER) were measured during each exercise. Energy expenditure was calculated from the steady state VO(2) adjusted for substrate metabolism using RER. Carbohydrate and fat oxidation were calculated from VO(2) and RER based on the assumption that protein breakdown contributes little to energy metabolism during exercise. RESULTS: Energy expenditure was greater (p<0.05) at 80 rev/min than at 40 rev/min. No difference was found between 40 and 60 rev/min and between 60 and 80 rev/min during both UE and LE. During LE, carbohydrate oxidation was also higher at 80 rev/min than at 40 rev/min, whereas no difference in fat oxidation was found among all three pedal rates. During UE, no speed related differences in either carbohydrate or fat utilisation were observed. CONCLUSIONS: Pedalling at a greater frequency helped to maximise energy expenditure during exercise using UE or LE despite an unchanging power output. Whereas contraction frequency affects energy expenditure similarly during both UE and LE, its impact on carbohydrate utilisation appears to be influenced by exercise modality or relative exercise intensity. PMID- 14751943 TI - The Football Association Medical Research Programme: an audit of injuries in professional football--analysis of hamstring injuries. AB - OBJECTIVE: To conduct a detailed analysis of hamstring injuries sustained in English professional football over two competitive seasons. METHODS: Club medical staff at 91 professional football clubs annotated player injuries over two seasons. A specific injury audit questionnaire was used together with a weekly form that documented each clubs' current injury status. RESULTS: Completed injury records for the two competitive seasons were obtained from 87% and 76% of the participating clubs respectively. Hamstring strains accounted for 12% of the total injuries over the two seasons with nearly half (53%) involving the biceps femoris. An average of five hamstring strains per club per season was observed. A total of 13 116 days and 2029 matches were missed because of hamstring strains, giving an average of 90 days and 15 matches missed per club per season. In 57% of cases, the injury occurred during running. Hamstring strains were most often observed during matches (62%) with an increase at the end of each half (p<0.01). Groups of players sustaining higher than expected rates of hamstring injury were Premiership (p<0.01) and outfield players (p<0.01), players of black ethnic origin (p<0.05), and players in the older age groups (p<0.01). Only 5% of hamstring strains underwent some form of diagnostic investigation. The reinjury rate for hamstring injury was 12%. CONCLUSION: Hamstring strains are common in football. In trying to reduce the number of initial and recurrent hamstring strains in football, prevention of initial injury is paramount. If injury does occur, the importance of differential diagnosis followed by the management of all causes of posterior thigh pain is emphasised. Clinical reasoning with treatment based on best available evidence is recommended. PMID- 14751944 TI - Clinical investigation of athletes with persistent fatigue and/or recurrent infections. AB - OBJECTIVE: To investigate whether underlying medical conditions contribute to the fatigue and high incidence of infections that can occur during repeated intense training. METHOD: Forty one competitive athletes (22 male, 19 female) with persistent fatigue and/or recurrent infections associated with performance decrements had a thorough medical examination and a series of clinical investigations to identify potential medical causes. RESULTS: Conditions with the potential to cause fatigue and/or recurrent infections were identified in 68% of the athletes. The most common were partial humoral immune deficiency (28%) and unresolved viral infections (27%). Non-fasting hypoglycaemia was common (28%). Other conditions included allergic disease (15%), new or poorly controlled asthma (13%), upper airway dysfunction (5%), sleep disorders (15%), iron depletion (3%), and one case of a thyroid disorder. A positive antinuclear antibody was detected in 21% of the athletes, without any clinical evidence of autoimmune disorders. Evidence of Epstein-Barr virus reactivation was detected in 22% of the athletes tested. CONCLUSIONS: Athletes with recurrent infections, fatigue, and associated poor performance may benefit from a thorough investigation of potentially reversible underlying medical conditions, especially when these conditions cause disruption to training and competition. Unresolved viral infections are not routinely assessed in elite athletes, but it may be worth considering in those experiencing fatigue and performing poorly. PMID- 14751945 TI - Performance enhanced headgear: a scientific approach to the development of protective headgear. AB - BACKGROUND: There is a continuing debate about the performance of protective headgear in rugby union, rugby league, and Australian rules football. OBJECTIVES: To examine the impact energy attenuation performance of foam that could be incorporated into headgear and examine the performance of prototypes of modified headgear. METHODS: Impact tests were conducted on polyethylene foams and protective headgear. Free fall drop tests with a rigid headform on to a flat rigid anvil were conducted. Resultant headform acceleration was measured. Means of the headform acceleration maxima for repeat tests were calculated. RESULTS: Tests on polyethylene foam indicated that an increase in thickness from 10 mm to 16 mm would improve headgear performance. These modifications were incorporated in part into two headgear models: the Albion Headpro and the Canterbury brand Body Armour honeycomb headgear. The headgear tests show that significant reductions in headform acceleration were achieved by increasing the foam density and thickness. Mean headform acceleration maxima for the 16 mm thick modified rugby headgear was about 25% of that of standard headgear for lateral impact 0.3 and 0.4 m drop heights and 27% for the centre front 0.3 m drop tests. At these impacts, the headform acceleration for the modified rugby headgear was below 200 g. CONCLUSIONS: Significant improvements in impact energy attenuation performance are possible with small design changes. Whether these are sufficient to reduce the rate or severity of concussion in rugby and Australian rules football can only be shown by formal prospective studies on the field. PMID- 14751946 TI - Effect of tai chi exercise on proprioception of ankle and knee joints in old people. AB - OBJECTIVES: To assess if tai chi, a traditional Chinese form of exercise, could improve proprioception in old people and if the effects of tai chi on proprioception are more evident than other exercise forms in the elderly. METHODS: By detecting the threshold of passive movement, ankle and knee joint kinaesthesis was measured in 21 elderly long term tai chi practitioners (TC group), 20 elderly long term swimmers/runners (S/R group), and 27 elderly sedentary controls (control group). RESULTS: Ankle joint kinaesthesis differed significantly among the three groups (p = 0.001). Subjects in the TC group could detect a significantly smaller amount of motion than those in the S/R group (p = 0.022) and control group (p = 0.001). No significant difference was found between the S/R group and the control group (p = 0.701). The threshold for detection of passive motion was significantly different in knee extension and flexion. For knee flexion, the TC group showed a significantly lower mean threshold for detection of passive motion than the control group (p = 0.026). There were no significant differences between the S/R group and control group (p = 0.312), or between the TC group and S/R group (p = 0.533). For knee extension, no significant difference was noted among the three groups (p = 0.597). CONCLUSIONS: The elderly people who regularly practiced tai chi not only showed better proprioception at the ankle and knee joints than sedentary controls, but also better ankle kinaesthesis than swimmers/runners. The large benefits of tai chi exercise on proprioception may result in the maintenance of balance control in older people. PMID- 14751947 TI - Effect of ultramarathon cycling on the heart rate in elite cyclists. AB - OBJECTIVES: To analyse the heart rate (HR) response and estimate the ultraendurance threshold-the optimum maintainable exercise intensity of ultraendurance cycling-in ultraendurance elite cyclists competing in the Race across the Alps. METHODS: HR monitoring was performed in 10 male elite cyclists during the first Race across the Alps in 2001 (distance: 525 km; cumulative altitude difference: 12 600 m) to investigate the exercise intensity of a cycle ultramarathon and the cardiopulmonary strains involved. Four different exercise intensities were defined as percentages of maximal HR (HR(max)) as follows: recovery HR (HR(re)), <70% of HR(max); moderate aerobic HR (HR(ma)), 70-80%; intense aerobic HR (HR(ia)), 80-90%; and high intensity HR (HR(hi)), >90%. RESULTS: All athletes investigated finished the competition. The mean racing time was 27 hours and 25 minutes, and the average speed was 18.6 km/h. The mean HR(max) was 186 beats/min, and the average value of measured HRs (HR(average)) was 126 beats/min resulting in a mean HR(average)/HR(max) ratio of 0.68, which probably corresponds to the ultraendurance threshold. The athletes spent 53% (14 hours 32 minutes) of total race time within HR(re), 25% (6 hours 51 minutes) within HR(ma), 19% (5 hours 13 minutes) within HR(ia), and only 3% (49 minutes) within HR(hi), which shows the exercise intensity to be predominantly moderate (HR(re) + HR(ma) = 78% or 21 hours 23 minutes). The HR response was influenced by the course profile as well as the duration. In all subjects, exercise intensity declined significantly during the race, as indicated by a decrease in HR(average)/HR(max) of 23% from 0.86 at the start to 0.66 at the end. CONCLUSIONS: A substantial decrease (10% every 10 hours) in the HR response is a general cardiovascular feature of ultramarathon cycling, suggesting that the ultraendurance threshold lies at about 70% of HR(max) in elite ultramarathon cyclists. PMID- 14751948 TI - A comparison of the sports safety policies and practices of community sports clubs during training and competition in northern Sydney, Australia. AB - OBJECTIVES: To compare the safety policies and practices reported to be adopted during training and competition by community sports clubs in northern Sydney, Australia. METHODS: This cross sectional study involved face to face interviews, using an 81 item extensively validated questionnaire, with representatives of 163 community netball, rugby league, rugby union, and soccer clubs (response rate 85%). The study was undertaken during the winter sports season of 2000. Two separate 14 item scales were developed to analyse the level of safety policy adoption and safety practice implementation during training and competition. The statistical analysis comprised descriptive and inferential analysis stratified by sport. RESULTS: The reliability of the scales was good: Cronbach's alpha = 0.70 (competition scale) to 0.81 (training scale). Significant differences were found between the safety scores for training and competition for all clubs (mean difference 11.2; 95% confidence interval (CI) 10.0 to 12.5) and for each of the four sports: netball (mean difference 14.9; 95% CI 12.6 to 17.2); rugby league (mean difference 10.3; 95% CI 7.1 to 13.6); rugby union (mean difference 9.4; 95% CI 7.1 to 11.7); and soccer (mean difference 8.4; 95% CI 6.5 to 10.3). CONCLUSIONS: The differences in the mean competition and training safety scores were significant for all sports. This indicates that safety policies were less often adopted and practices less often implemented during training than during competition. As injuries do occur at training, and sports participants often spend considerably more time training than competing, sporting bodies should consider whether the safety policies and practices adopted and implemented at training are adequate. PMID- 14751949 TI - Evaluation of isokinetic force production and associated muscle activity in the scapular rotators during a protraction-retraction movement in overhead athletes with impingement symptoms. AB - OBJECTIVES: To determine if the muscle force and electromyographic activity in the scapular rotators of overhead athletes with impingement symptoms showed differences between the injured and non-injured sides. METHODS: Isokinetic peak force was evaluated during protraction and retraction of the shoulder girdle, with simultaneous recording of electromyographic activity of the three trapezius muscles and the serratus anterior muscle, in 19 overhead athletes with impingement symptoms. RESULTS: Paired t tests showed significantly lower peak force during isokinetic protraction at high velocity (p<0.05), a significantly lower protraction/retraction ratio (p<0.01), and significantly lower electromyographic activity in the lower trapezius muscle during isokinetic retraction on the injured side than on the non-injured side (p<0.05). CONCLUSION: These results confirm that patients with impingement symptoms show abnormal muscle performance at the scapulothoracic joint. PMID- 14751950 TI - Prevalence of temporomandibular dysfunction in a group of scuba divers. AB - BACKGROUND: Temporomandibular dysfunction (TMD) has been reported to be a common problem in divers, with a prevalence of up to 68%. No evidence for this is available. OBJECTIVE: To investigate the prevalence of TMD in divers. METHOD: Sixty three subjects were asked to retrospectively complete a questionnaire on symptoms of TMD after diving in warm and cold water areas and in daily life. RESULTS: The prevalence of TMD was greater in female divers. The prevalence of TMD while diving was about 26%, comparable to that experienced in daily life. CONCLUSION: Improvements in mouthpiece design and lighter demand valves mean that TMD is now probably exacerbated by diving rather than caused by it. PMID- 14751951 TI - Effect of physiotherapy attendance on outcome after anterior cruciate ligament reconstruction: a pilot study. AB - BACKGROUND: In many centres patients are routinely referred for physiotherapy after anterior cruciate ligament (ACL) reconstruction. However, to date the role and amount of supervised physiotherapy required has not been clearly established. OBJECTIVE: To establish whether there was any difference in outcome between a group of patients who attended physiotherapy regularly after ACL reconstruction and a group who attended only infrequently. METHODS: Ten patients who had attended physiotherapy infrequently (mean 1.9 visits) during the first six months after ACL reconstructive surgery were matched for age, sex, graft type, and activity level and occupation before injury with 10 patients who had attended physiotherapy regularly (mean 26.5 visits). Outcome was assessed at 12 months using the Cincinnati knee rating system and the IKDC form. RESULTS: Compared with the regular physiotherapy group, patients in the minimal physiotherapy group had fewer symptoms (mean Cincinnati symptom score 46.2 v 43.4, p = 0.045). There was also a trend towards higher overall Cincinnati knee scores in the minimal physiotherapy group (mean 93.7 v 87.3, p = 0.06) but no difference in IKDC ratings. CONCLUSION: These preliminary results indicate that some patients who choose to attend physiotherapy on a very limited basis after ACL reconstruction can achieve satisfactory, if not better, outcomes than patients who attend physiotherapy regularly. PMID- 14751952 TI - Fatal head injury from boxing: a case report from Greece. AB - The case is described of a fatal injury to a young amateur boxer. The number of deaths from boxing have decreased in recent years. However, new rules devised to promote safety, such as limitation of the number of rounds, mandatory use of safety head guards, and better medical control during the match, seem to be of limited value in preventing severe head injuries from boxing. PMID- 14751953 TI - Deep cutting injury from the edges of a snowboard. AB - A laceration deep enough to reach the bone occurs very rarely in skiing or snowboarding. Two such cases are presented here. In one case, the popliteal fossa of a skier was cut during a collision with a snowboarder. All structures posterior to the knee were severed and the leg became ischaemic. The other case was of a snowboarder who sustained a deep cut to the distal forearm during landing after a jump, resulting in a "spaghetti wrist". PMID- 14751954 TI - Exercise induced neurally mediated syncope in an elite rower: a treatment dilemma. PMID- 14751955 TI - Effect of prolonged exercise in a hypoxic environment on cardiac function and cardiac troponin T. AB - BACKGROUND: Exercise induced cardiac fatigue has recently been observed after prolonged exercise. A moderate to high altitude has been suggested as a possible stimulus in the genesis of such cardiac fatigue. OBJECTIVE: To investigate if exercise induced cardiac fatigue and or cardiac damage occurs after prolonged exercise in a hypoxic environment. METHODS: Eight trained male triathletes volunteered for the study. Each completed two 50 mile cycle trials, randomly assigned from normobaric normoxia and normobaric hypoxia (15% FIO(2)). Echocardiographic assessment and whole blood collection was completed before, immediately after, and 24 hours after exercise. Left ventricular systolic and diastolic functional variables were calculated, and serum was analysed for cardiac troponin T. Results were analysed using a two way repeated measures analysis of variance, with alpha set at 0.05. RESULTS: No significant differences were observed in either systolic or diastolic function across time or between trials. Cardiac troponin T was detected in one subject immediately after exercise in the normobaric hypoxic trial. CONCLUSIONS: A 50 mile cycle trial in either normobaric normoxia or normobaric hypoxia does not induce exercise induced cardiac fatigue. Some people, however, may exhibit minimal cardiac damage after exercise in normobaric hypoxia. The clinical significance of this is yet to be elucidated. PMID- 14751956 TI - Strategies for prevention of soccer related injuries: a systematic review. AB - OBJECTIVES: To examine evidence on the effectiveness of current injury prevention strategies in soccer, determine the applicability of the evidence to children and youth, and make recommendations on policy, programming, and future research. METHODS: Standard systematic review methodology was modified and adopted for this review. Research questions and relevance criteria were developed a priori. Potentially relevant studies were located through electronic and hand searches. Articles were assessed for relevance and quality by two independent assessors, and the results of relevant articles were abstracted and synthesised. RESULTS: A total of 44 potentially relevant articles from electronic (n = 37) and hand (n = 7) searches yielded four that met inclusion criteria. These four studies addressed a range of intervention strategies and varied with respect to results and quality of evidence. CONCLUSIONS: Some of the strategies look promising but lack adequate evaluation or require further research among younger players. Practice, policy, and research recommendations are provided as a result of the synthesis. PMID- 14751957 TI - Use of tissue adhesives in sport? A new application in international ice hockey. AB - In the tough, competitive environment of international ice hockey, it is vital that a player who sustains a minor facial laceration is returned to play as soon as possible. A method of wound closure that is fast to apply, water resistant, and effective was sought. Dermabond, a cyanoacrylate tissue adhesive, was selected for use during the 2002 International Ice Hockey Federation World Championships for the closure of selected facial wounds. The wounds were all closed using prescribed aseptic techniques. The results are presented and discussed. The tissue adhesive met the criteria set out. PMID- 14751958 TI - Raised concentrations of C reactive protein in anabolic steroid using bodybuilders. AB - OBJECTIVE: To examine levels of C reactive protein in users of anabolic androgenic steroids (AAS) compared with age matched control groups consisting of AAS using (but abstinent)/resistance trained and non-drug using/sedentary controls. METHOD: Subjects included AAS using bodybuilders (n = 10); bodybuilders who denied AAS use (n = 10); sedentary controls (n = 8). Venous blood was sampled, from which serum concentrations of C reactive protein, male sex hormones, and cardiac troponin T were determined. RESULTS: A significantly altered hormonal profile in the AAS using group provided indirect confirmation of AAS use. C reactive protein concentrations were significantly (p<0.05) higher in the AAS using bodybuilders. There was no relation between C reactive protein and cardiac troponin T. CONCLUSION: AAS using bodybuilders had significantly higher C reactive protein concentrations, indicating a greater propensity to develop peripheral arterial disease. PMID- 14751959 TI - Blood boosting. AB - This article reviews the history, technique, effects, side effects, and detection of blood boosting. It also considers whether or not this particular performance enhancement technique is a thing of the past or a continuing form of abuse among athletes. PMID- 14751960 TI - Scuba diving can induce stress of the temporomandibular joint leading to headache. PMID- 14751961 TI - Metabolic risks of completing Giro, Tour, and Vuelta in the same season. PMID- 14751963 TI - Modeling the relationship between LVAD support time and gene expression changes in the human heart by penalized partial least squares. AB - MOTIVATION: Heart failure affects more than 20 million people in the world. Heart transplantation is the most effective therapy, but the number of eligible patients far outweighs the number of available donor hearts. The left mechanical ventricular assist device (LVAD) has been developed as a successful substitution therapy that aids the failing ventricle while a patient is waiting for the donor heart. We obtained genomics data from paired human heart samples harvested at the time of LVAD implant and explant. The heart failure patients in our study were supported by the LVAD for various periods of time. The goal of this study is to model the relationship between the time of LVAD support and gene expression changes. RESULTS: To serve the purpose, we propose a novel penalized partial least squares (PPLS) method to build a regression model. Compared with partial least squares and Breiman's random forest method, PPLS gives the best prediction results for the LVAD data. PMID- 14751964 TI - Mapping Gene Ontology to proteins based on protein-protein interaction data. AB - MOTIVATION: Gene Ontology (GO) consortium provides structural description of protein function that is used as a common language for gene annotation in many organisms. Large-scale techniques have generated many valuable protein-protein interaction datasets that are useful for the study of protein function. Combining both GO and protein-protein interaction data allows the prediction of function for unknown proteins. RESULT: We apply a Markov random field method to the prediction of yeast protein function based on multiple protein-protein interaction datasets. We assign function to unknown proteins with a probability representing the confidence of this prediction. The functions are based on three general categories of cellular component, molecular function and biological process defined in GO. The yeast proteins are defined in the Saccharomyces Genome Database (SGD). The protein-protein interaction datasets are obtained from the Munich Information Center for Protein Sequences (MIPS), including physical interactions and genetic interactions. The efficiency of our prediction is measured by applying the leave-one-out validation procedure to a functional path matching scheme, which compares the prediction with the GO description of a protein's function from the abstract level to the detailed level along the GO structure. For biological process, the leave-one-out validation procedure shows 52% precision and recall of our method, much better than that of the simple guilty-by-association methods. PMID- 14751965 TI - Predicting the linkage sites in glycoproteins using bio-basis function neural network. AB - MOTIVATION: Although, it is known that O-glycosidically linked oligosaccharides are commonly conjugated to a serine, threonine or hydroxylysine residue of the polypeptide, the chemical nature of the anchoring monosaccharide and the size of the oligosaccharide unit varies. Among different types, O-linked or mucin-type oligosaccharides are intimately involved in the secretion of proteins, be they enzymes, hormones or structural glycoproteins. Knowledge of the linkage sites in glycoproteins is critical to the design of specific and efficient inhibitors against the enzyme to catalyse the formation of the carbohydrate-peptide linkage. RESULTS: We present a method for predicting the linkage sites in O-linked glycoproteins using bio-basis function neural networks. The mean prediction accuracy of this method is 91.15 +/- 2.75% while it is 82.28 +/- 6.45% using back propagation neural networks. Importantly, this method has significantly reduced the CPU time for modelling. PMID- 14751966 TI - Gene Unscrambler for detangling scrambled genes in ciliates. AB - Scrambled genes are surprisingly common in some species of ciliates. Until now there was no software available to analyze automatically these genes. We present here a program that can automatically align the macronuclear and micronuclear forms of a gene, outputting the location of the macronuclear destined segments and pointer sequences. AVAILABILITY: A web version of the program is available free of charge and can be accessed at http://oxytricha.princeton.edu/GeneUnscrambler.htm PMID- 14751967 TI - ReDiT: Repeat Discrepancy Tagger--a shotgun assembly finishing aid. AB - Finishing, i.e. gap closure and editing, is the most time-consuming part of genome sequencing. Repeated sequences together with sequencing errors complicate the assembly and often result in misassemblies that are difficult to correct. Repeat Discrepancy Tagger (ReDiT) is a tool designed to aid in the finishing step. This software processes assembly results produced by any fragment assembly program that outputs ace files. The input sequences are analyzed to determine possible differences between repeated sequences. The output is written as tags in an ace file that can be viewed by, e.g. the Consed sequence editor. AVAILABILITY: The ReDiT program is freely available at http://web.cgb.ki.se/redit PMID- 14751968 TI - Alu repeat analysis in the complete human genome: trends and variations with respect to genomic composition. AB - MOTIVATION: Transposon-derived Alu repeats are exclusively associated with primate genomes. They have gained considerable importance in the recent times with evidence of their involvement in various aspects of gene regulation, e.g. alternative splicing, nucleosome positioning, CpG methylation, binding sites for transcription factors and hormone receptors, etc. The objective of this study is to investigate the factors that influence the distribution of Alu repeat elements in the human genome. Such analysis is expected to yield insights into various aspects of gene regulation in primates. RESULTS: Analysis of Alu repeat distribution for the human genome build 32 (released in January 2003) reveals that they occupy nearly one-tenth portion of the sequenced regions. Huge variations in Alu frequencies were seen across the genome with chromosome 19 being the most and chromosome Y being the least Alu dense chromosomes. The highlights of the analysis are as follows: (1). three-fourth of the total genes in the genome are associated with Alus. (2). Alu density is higher in genes as compared with intergenic regions in all the chromosomes except 19 and 22. (3). Alu density in human genome is highly correlated with GC content, gene density and intron density with GC content being major deterministic factor compared with other two. (4). Alu densities were correlated more with gene density than intron density indicating the insertion of Alus in untranslated regions of exons. PMID- 14751969 TI - Modeling within-motif dependence for transcription factor binding site predictions. AB - MOTIVATION: The position-specific weight matrix (PWM) model, which assumes that each position in the DNA site contributes independently to the overall protein DNA interaction, has been the primary means to describe transcription factor binding site motifs. Recent biological experiments, however, suggest that there exists interdependence among positions in the binding sites. In order to exploit this interdependence to aid motif discovery, we extend the PWM model to include pairs of correlated positions and design a Markov chain Monte Carlo algorithm to sample in the model space. We then combine the model sampling step with the Gibbs sampling framework for de novo motif discoveries. RESULTS: Testing on experimentally validated binding sites, we find that about 25% of the transcription factor binding motifs show significant within-site position correlations, and 80% of these motif models can be improved by considering the correlated positions. Using both simulated data and real promoter sequences, we show that the new de novo motif-finding algorithm can infer the true correlated position pairs accurately and is more precise in finding putative transcription factor binding sites than the standard Gibbs sampling algorithms. PMID- 14751970 TI - Gaussian mixture clustering and imputation of microarray data. AB - MOTIVATION: In microarray experiments, missing entries arise from blemishes on the chips. In large-scale studies, virtually every chip contains some missing entries and more than 90% of the genes are affected. Many analysis methods require a full set of data. Either those genes with missing entries are excluded, or the missing entries are filled with estimates prior to the analyses. This study compares methods of missing value estimation. RESULTS: Two evaluation metrics of imputation accuracy are employed. First, the root mean squared error measures the difference between the true values and the imputed values. Second, the number of mis-clustered genes measures the difference between clustering with true values and that with imputed values; it examines the bias introduced by imputation to clustering. The Gaussian mixture clustering with model averaging imputation is superior to all other imputation methods, according to both evaluation metrics, on both time-series (correlated) and non-time series (uncorrelated) data sets. PMID- 14751971 TI - External control in Markovian genetic regulatory networks: the imperfect information case. AB - Probabilistic Boolean Networks, which form a subclass of Markovian Genetic Regulatory Networks, have been recently introduced as a rule-based paradigm for modeling gene regulatory networks. In an earlier paper, we introduced external control into Markovian Genetic Regulatory networks. More precisely, given a Markovian genetic regulatory network whose state transition probabilities depend on an external (control) variable, a Dynamic Programming-based procedure was developed by which one could choose the sequence of control actions that minimized a given performance index over a finite number of steps. The control algorithm of that paper, however, could be implemented only when one had perfect knowledge of the states of the Markov Chain. This paper presents a control strategy that can be implemented in the imperfect information case, and makes use of the available measurements which are assumed to be probabilistically related to the states of the underlying Markov Chain. PMID- 14751972 TI - mdclust--exploratory microarray analysis by multidimensional clustering. AB - MOTIVATION: Unsupervised clustering of microarray data may detect potentially important, but not obvious characteristics of samples, for instance subgroups of diagnoses with distinct gene profiles or systematic errors in experimentation. RESULTS: Multidimensional clustering (mdclust) is a method, which identifies sets of sample clusters and associated genes. It applies iteratively two-means clustering and score-based gene selection. For any phenotype variable best matching sets of clusters can be selected. This provides a method to identify gene-phenotype associations, suited even for settings with a large number of phenotype variables. An optional model based discriminant step may reduce further the number of selected genes. PMID- 14751973 TI - The Hera database and its use in the characterization of endoplasmic reticulum proteins. AB - MOTIVATION: Information concerning endoplasmic reticulum (ER) proteins is widely dispersed and cannot be easily and rapidly processed by the biological community. We present a comprehensive database of human ER proteins, called Human ER Apercu (Hera). The Hera database was constructed by exhaustively searching through public databases and the scientific literature for ER proteins. RESULTS: Hera was used for the analysis of characteristics common to all human ER proteins. Our results show that a high proportion of ER proteins (59%) have at least one transmembrane domain and display physical characteristics consistent with this observation. In addition, one-third of ER proteins contain known ER retrieval or retention signals and 70% of ER proteins contain a signal peptide or anchor. Finally, 85% of ER proteins contain at least one InterPro motif. The most abundant InterPro motifs in ER proteins represent many of the most well characterized functions of the ER. PMID- 14751974 TI - Finding coexpressed genes in counts-based data: an improved measure with validation experiments. AB - MOTIVATION: Expressed sequence tag (EST) data reflects variation in gene expression, but previous methods for finding coexpressed genes in EST data are subject to bias and vastly overstate the statistical significance of putatively coexpressed genes. RESULTS: We introduce a new method (LNP) that reports reasonable p-values and also detects more biological relationships in human dbEST than do previous methods. In simulations with human dbEST library sizes, previous methods report p-values as low as 10(-30) on 1/1000 uncorrelated pairs, while LNP reports significance correctly. We validate the analysis on real human genes by comparing coexpressed pairs to gene ontology annotations and find that LNP is more sensitive than the three previous methods. We also find a small but statistically significant level of coexpression between interacting proteins relative to randomized controls. The LNP method is based on a log-normal prior on the distribution of expression levels. PMID- 14751976 TI - ClusterControl: a web interface for distributing and monitoring bioinformatics applications on a Linux cluster. AB - ClusterControl is a web interface to simplify distributing and monitoring bioinformatics applications on Linux cluster systems. We have developed a modular concept that enables integration of command line oriented program into the application framework of ClusterControl. The systems facilitate integration of different applications accessed through one interface and executed on a distributed cluster system. The package is based on freely available technologies like Apache as web server, PHP as server-side scripting language and OpenPBS as queuing system and is available free of charge for academic and non-profit institutions. AVAILABILITY: http://genome.tugraz.at/Software/ClusterControl PMID- 14751975 TI - Visualization of near-optimal sequence alignments. AB - MOTIVATION: Mathematically optimal alignments do not always properly align active site residues or well-recognized structural elements. Most near-optimal sequence alignment algorithms display alternative alignment paths, rather than the conventional residue-by-residue pairwise alignment. Typically, these methods do not provide mechanisms for finding effectively the most biologically meaningful alignment in the potentially large set of options. RESULTS: We have developed Web based software that displays near optimal or alternative alignments of two protein or DNA sequences as a continuous moving picture. A WWW interface to a C++ program generates near optimal alignments, which are sent to a Java Applet, which displays them in a series of alignment frames. The Applet aligns residues so that consistently aligned regions remain at a fixed position on the display, while variable regions move. The display can be stopped to examine alignment details. PMID- 14751977 TI - Identification of optimal classification functions for biological sample and state discrimination from metabolic profiling data. AB - MOTIVATIONS: Classification of biological samples for diagnostic purposes is a difficult task because of the many decisions involved on the number, type and functional manipulations of the input variables. This study presents a generally applicable strategy for systematic formulation of optimal diagnostic indexes. To this end, we develop a novel set of computational tools by integrating regression optimization, stepwise variable selection and cross-validation algorithms. RESULTS: The proposed discrimination methodology was applied to plasma and tissue (liver) metabolic profiling data describing the time progression of liver dysfunction in a rat model of acute hepatic failure generated by d-galactosamine (GalN) injection. From the plasma data, our methodology identified seven (out of a total of 23) metabolites, and the corresponding transform functions, as the best inputs to the optimal diagnostic index. This index showed better time resolution and increased noise robustness compared with an existing metabolic index, Fischer's BCAA/AAA molar ratio, as well as indexes generated using other commonly used discriminant analysis tools. Comparison of plasma and liver indexes found two consensus metabolites, lactate and glucose, which implicate glycolysis and/or gluconeogenesis in mediating the metabolic effects of GalN. PMID- 14751980 TI - Effects of choice of DNA sequence model structure on gene identification accuracy. AB - MOTIVATION: Markov chain models of DNA sequences have frequently been used in gene finding algorithms. Performance of the algorithm critically depends on the model structure and parameters. Still, the issue of choosing the model structure has not been studied with sufficient attention. RESULTS: We have assessed performance of several types of Markov chain models, both fixed order (FO) models and models with interpolation, within the framework of the GeneMark algorithm. The performance was measured in two ways: (i) the accuracy of detection of protein-coding potential in artificial DNA sequences and (ii) the accuracy of identifying genes in real prokaryotic genomes. We observed that the models built by deleted interpolation (DI) slightly outperformed other models in detecting protein-coding potential in artificial DNA sequences with GC content in the medium range and also in detecting genes in real genomes with medium GC content. For artificial and real genomic DNA with high or low GC content, we observed that the models built by DI were in some cases slightly outperformed by FO models. PMID- 14751981 TI - Large-scale analysis of non-synonymous coding region single nucleotide polymorphisms. AB - MOTIVATION: Single nucleotide polymorphisms (SNPs) are the most common form of genetic variant in humans. SNPs causing amino acid substitutions are of particular interest as candidates for loci affecting susceptibility to complex diseases, such as diabetes and hypertension. To efficiently screen SNPs for disease association, it is important to distinguish neutral variants from deleterious ones. RESULTS: We describe the use of Pfam protein motif models and the HMMER program to predict whether amino acid changes in conserved domains are likely to affect protein function. We find that the magnitude of the change in the HMMER E-value caused by an amino acid substitution is a good predictor of whether it is deleterious. We provide internet-accessible display tools for a genomewide collection of SNPs, including 7391 distinct non-synonymous coding region SNPs in 2683 genes. AVAILABILITY: http://lpgws.nci.nih.gov/cgi bin/GeneViewer.cgi PMID- 14751983 TI - Measuring the similarity of protein structures by means of the universal similarity metric. AB - MOTIVATION: As an increasing number of protein structures become available, the need for algorithms that can quantify the similarity between protein structures increases as well. Thus, the comparison of proteins' structures, and their clustering accordingly to a given similarity measure, is at the core of today's biomedical research. In this paper, we show how an algorithmic information theory inspired Universal Similarity Metric (USM) can be used to calculate similarities between protein pairs. The method, besides being theoretically supported, is surprisingly simple to implement and computationally efficient. RESULTS: Structural similarity between proteins in four different datasets was measured using the USM. The sample employed represented alpha, beta, alpha-beta, tim barrel, globins and serpine protein types. The use of the proposed metric allows for a correct measurement of similarity and classification of the proteins in the four datasets. AVAILABILITY: All the scripts and programs used for the preparation of this paper are available at http://www.cs.nott.ac.uk/~nxk/USM/protocol.html. In that web-page the reader will find a brief description on how to use the various scripts and programs. PMID- 14751982 TI - Database model and specification of GermOnline Release 2.0, a cross-species community annotation knowledgebase on germ cell differentiation. AB - GermOnline is a web-accessible relational database that enables life scientists to make a significant and sustained contribution to the annotation of genes relevant for the fields of mitosis, meiosis, germ line development and gametogenesis across species. This novel approach to genome annotation includes a platform for knowledge submission and curation as well as microarray data storage and visualization hosted by a global network of servers. AVAILABILITY: The database is accessible at http://www.germonline.org/. For convenient world-wide access we have set up a network of servers in Europe (http://germonline.unibas.ch/; http://germonline.igh.cnrs.fr/), Japan (http://germonline.biochem.s.u-tokyo.ac.jp/) and USA (http://germonline.yeastgenome.org/). SUPPLEMENTARY INFORMATION: Extended documentation of the database is available through the link 'About GermOnline' at the websites. PMID- 14751984 TI - On the significance of sequence alignments when using multiple scoring matrices. AB - MOTIVATION: Pairwise local sequence alignment is commonly used to search data bases for sequences related to some query sequence. Alignments are obtained using a scoring matrix that takes into account the different frequencies of occurrence of the various types of amino acid substitutions. Software like BLAST provides the user with a set of scoring matrices available to choose from, and in the literature it is sometimes recommended to try several scoring matrices on the sequences of interest. The significance of an alignment is usually assessed by looking at E-values and p-values. While sequence lengths and data base sizes enter the standard calculations of significance, it is much less common to take the use of several scoring matrices on the same sequences into account. Altschul proposed corrections of the p-value that account for the simultaneous use of an infinite number of PAM matrices. Here we consider the more realistic situation where the user may choose from a finite set of popular PAM and BLOSUM matrices, in particular the ones available in BLAST. It turns out that the significance of a result can be considerably overestimated, if a set of substitution matrices is used in an alignment problem and the most significant alignment is then quoted. RESULTS: Based on extensive simulations, we study the multiple testing problem that occurs when several scoring matrices for local sequence alignment are used. We consider a simple Bonferroni correction of the p-values and investigate its accuracy. Finally, we propose a more accurate correction based on extreme value distributions fitted to the maximum of the normalized scores obtained from different scoring matrices. For various sets of matrices we provide correction factors which can be easily applied to adjust p- and E-values reported by software packages. PMID- 14751986 TI - The PCR suite. AB - The web application PCR Suite is an extension of the primer design program Primer3. It allows the design of primer sets encompassing single nucleotide polymorphisms, all exons of a single gene, all open reading frames in a list of cDNAs or the creation of overlapping PCR products. PMID- 14751985 TI - Using functional and organizational information to improve genome-wide computational prediction of transcription units on pathway-genome databases. AB - MOTIVATION: The prediction of transcription units (TUs, which are similar to operons) is an important problem that has been tackled using many different approaches. The availability of complete microbial genomes has made genome-wide TU predictions possible. Pathway-genome databases (PGDBs) add metabolic and other organizational (i.e. protein complexes) information to the annotated genome, and are able to capture TU organization information. These characteristics of PGDBs make them a suitable framework for the development and implementation of TU predictors. RESULTS: We implemented a TU predictor that uses only intergenic distance and functional classification of genes to predict TU boundaries, and applied it to EcoCyc, our PGDB of Escherichia coli. To this original predictor, we added information on metabolic pathways, protein complexes and transporters, all readily available in EcoCyc, in order to generate an enhanced predictor. The enhanced predictor correctly predicted 80% of the known E.coli TUs (69% of the known operons), a moderate improvement over the original predictor's performance (75% of TUs and 65% of operons correctly predicted), demonstrating that the extra information available in the PGDB does indeed improve prediction performance. Performance of this E.coli-based predictor on a genome other than that of E.coli was tested on BsubCyc, our computationally generated PGDB for Bacillus subtilis, for which a set of 100 known operons is available. Prediction accuracy decreased substantially (46% of the known operons correctly predicted). This was due in part to missing information in BsubCyc, which prevented full use of the predictor's features. The augmented predictor has been implemented as part of our Pathway Tools software suite, and can be used to populate a PGDB with predicted TUs. AVAILABILITY: The TU predictor is included in version 7.0 of the Pathway Tools software suite. Pathway Tools 7.0 is available free of charge to academic institutions and for a fee to commercial enterprises. It runs on Sun Solaris 8, Linux and Windows. TUs predicted on the Caulobacter crescentus and Mycobacterium tuberculosis (H37Rv) genomes are available in our CauloCyc and MtbrvCyc databases, available at the BioCyc web site (http://biocyc.org). To obtain version 7.0 of Pathway Tools, follow the directions in our web site, http://biocyc.org/download.shtml. PMID- 14751987 TI - Genotyping of single nucleotide polymorphism using model-based clustering. AB - MOTIVATION: Single nucleotide polymorphisms have been investigated as biological markers and the representative high-throughput genotyping method is a combination of the Invader assay and a statistical clustering method. A typical statistical clustering method is the k-means method, but it often fails because of the lack of flexibility. An alternative fast and reliable method is therefore desirable. RESULTS: This paper proposes a model-based clustering method using a normal mixture model and a well-conceived penalized likelihood. The proposed method can judge unclear genotypings to be re-examined and also work well even when the number of clusters is unknown. Some results are illustrated and then satisfactory genotypings are shown. Even when the conventional maximum likelihood method and the typical k-means clustering method failed, the proposed method succeeded. PMID- 14751988 TI - Java-based application framework for visualization of gene regulatory region annotations. AB - MOTIVATION: The genome sequences of several organisms are either complete, or being sequenced. Each genome needs to be integrated with various types of annotations, e.g. locations of genes, promoters and other functional elements such as transcriptional regulatory elements. A robust application framework will be useful for developing web-based applications to visualize various genome annotations. RESULTS: We developed genome data visualization toolkit (GDVTK) as an application framework that consists of a set of data structures and core classes, using Java technology. GDVTK is a sound framework for developing web based applications to present the gene regulatory region annotations in visual form. The current version of GDVTK consists of eight packages and 38 Java classes that are portable, reusable and extensible for plugging in new data sources and models. We implemented GDVTK for visualization of promoter annotations in Mammalian Promoter Database (MPromDb), a web-based gene-regulatory information server. AVAILABILITY: GDVTK is available under GNU general public license. Source code and software documentation can be found at the URL http://bioinformatics.med.ohio-state.edu/GDVTK. PMID- 14751989 TI - Bio-support vector machines for computational proteomics. AB - MOTIVATION: One of the most important issues in computational proteomics is to produce a prediction model for the classification or annotation of biological function of novel protein sequences. In order to improve the prediction accuracy, much attention has been paid to the improvement of the performance of the algorithms used, few is for solving the fundamental issue, namely, amino acid encoding as most existing pattern recognition algorithms are unable to recognize amino acids in protein sequences. Importantly, the most commonly used amino acid encoding method has the flaw that leads to large computational cost and recognition bias. RESULTS: By replacing kernel functions of support vector machines (SVMs) with amino acid similarity measurement matrices, we have modified SVMs, a new type of pattern recognition algorithm for analysing protein sequences, particularly for proteolytic cleavage site prediction. We refer to the modified SVMs as bio-support vector machine. When applied to the prediction of HIV protease cleavage sites, the new method has shown a remarkable advantage in reducing the model complexity and enhancing the model robustness. PMID- 14751990 TI - GENIUS II: a high-throughput database system for linking ORFs in complete genomes to known protein three-dimensional structures. AB - GENIUS II is an automated database system in which open reading frames (ORFs) in complete genomes are assigned to known protein three-dimensional (3D) structures. The system uses the multiple intermediate sequence search method in which query and target sequences are linked by intermediate sequences gathered by PSI-BLAST search. By applying the system to 129 complete genomes, 43.8% on average of the ORFs in the genomes were assigned to known 3D structures and the results are available for free at GENIUS II web site. PMID- 14751991 TI - Dominant spectral component analysis for transcriptional regulations using microarray time-series data. AB - MOTIVATION: Microarray time-series data provides us a possible means for identification of transcriptional regulation relationships among genes. Currently, the most commonly used method in determining whether or not two genes have a potential regulatory relationship is to measure their expressional similarity using Pearson's correlation coefficient. Although this traditional correlation method has been successfully applied to find functionally correlated genes, it does have many limitations. In the hope of overcoming such circumstances and getting more insights into the transcriptional regulatory issue, we propose an autoregressive (AR)-based technique for detection of potential regulated gene pairs from time-series microarray measurements. RESULTS: We use the well-known AR modeling technique to characterize temporal gene expression data from the Spellman's alpha-synchronized yeast cell-cycle experiment. In this method, time-series expression profiles are decomposed into spectral components and correlations between profiles are then computed in a component-wise sense. We show how these component-wise correlations reveal possible regulatory relationships. Our technique is applied on known transcriptional regulations and is able to identify many of those missed by the traditional correlation method. PMID- 14751992 TI - Regression trees for regulatory element identification. AB - MOTIVATION: The transcription of a gene is largely determined by short sequence motifs that serve as binding sites for transcription factors. Recent findings suggest direct relationships between the motifs and gene expression levels. In this work, we present a method for identifying regulatory motifs. Our method makes use of tree-based techniques for recovering the relationships between motifs and gene expression levels. RESULTS: We treat regulatory motifs and gene expression levels as predictor variables and responses, respectively, and use a regression tree model to identify the structural relationships between them. The regression tree methodology is extended to handle responses from multiple experiments by modifying the split function. The significance of regulatory elements is determined by analyzing tree structures and using a variable importance measure. When applied to two data sets of the yeast Saccharomyces cerevisiae, the method successfully identifies most of the regulatory motifs that are known to control gene transcription under the given experimental conditions, and suggests several new putative motifs. Analysis of the tree structures also reconfirms several pairs of motifs that are known to regulate gene transcription in combination. AVAILABILITY: http://if.kaist.ac.kr/~phuong/RegTree PMID- 14751993 TI - STAM: simple transmembrane alignment method. AB - MOTIVATION: The database of transmembrane protein (TMP) structures is still very small. At the same time, more and more TMP sequences are being determined. Molecular modeling is an interim answer that may bridge the gap between the two databases. The first step in homology modeling is to achieve a good alignment between the target sequences and the template structure. However, since most algorithms to obtain the alignments were constructed with data derived from globular proteins, they perform poorly when applied to TMPs. In our application, we automate the alignment procedure and design it specifically for TMP. We first identify segments likely to form transmembrane alpha-helices. We then apply different sets of criteria for transmembrane and non-transmembrane segments. For example, the penalty for insertion/deletions in the transmembrane segments is much higher than that of a penalty in the loop region. Different substitution matrices are used since the frequencies of occurrence of the various amino acids differ for transmembrane segments and water-soluble domains. RESULTS: This program leads to better models since it does not treat the protein as a single entity with the same properties, but accounts for the different physical properties of the various segments. STAM is the first multisequence alignment program that is directly targeted at transmembrane proteins. AVAILABILITY: Source code and installation package are available on request from the authors. Web access is currently implemented. PMID- 14751994 TI - Predicting protein structure classes from function predictions. AB - MOTIVATION: We introduce a new approach to using the information contained in sequence-to-function prediction data in order to recognize protein template classes, a critical step in predicting protein structure. The data on which our method is based comprise probabilities of functional categories; for given query sequences these probabilities are obtained by a neural net that has previously been trained on a variety of functionally important features. On a training set of sequences we assess the relevance of individual functional categories for identifying a given structural family. Using a combination of the most relevant categories, the likelihood of a query sequence to belong to a specific family can be estimated. RESULTS: The performance of the method is evaluated using cross validation. For a fixed structural family and for every sequence, a score is calculated that measures the evidence for family membership. Even for structural families of small size, family members receive significantly higher scores. For some examples, we show that the relevant functional features identified by this method are biologically meaningful. The proposed approach can be used to improve existing sequence-to-structure prediction methods. AVAILABILITY: Matlab code is available on request from the authors. The data are available at http://www.mpisb.mpg.de/~sommer/Fun2Struc/ PMID- 14751995 TI - Reproducibility of SELDI-TOF protein patterns in serum: comparing datasets from different experiments. AB - MOTIVATION: There has been much interest in using patterns derived from surface enhanced laser desorption and ionization (SELDI) protein mass spectra from serum to differentiate samples from patients both with and without disease. Such patterns have been used without identification of the underlying proteins responsible. However, there are questions as to the stability of this procedure over multiple experiments. RESULTS: We compared SELDI proteomic spectra from serum from three experiments by the same group on separating ovarian cancer from normal tissue. These spectra are available on the web at http://clinicalproteomics.steem.com. In general, the results were not reproducible across experiments. Baseline correction prevents reproduction of the results for two of the experiments. In one experiment, there is evidence of a major shift in protocol mid-experiment which could bias the results. In another, structure in the noise regions of the spectra allows us to distinguish normal from cancer, suggesting that the normals and cancers were processed differently. Sets of features found to discriminate well in one experiment do not generalize to other experiments. Finally, the mass calibration in all three experiments appears suspect. Taken together, these and other concerns suggest that much of the structure uncovered in these experiments could be due to artifacts of sample processing, not to the underlying biology of cancer. We provide some guidelines for design and analysis in experiments like these to ensure better reproducible, biologically meaningfully results. AVAILABILITY: The MATLAB and Perl code used in our analyses is available at http://bioinformatics.mdanderson.org PMID- 14751996 TI - Quality of alignment comparison by COMPASS improves with inclusion of diverse confident homologs. AB - MOTIVATION: Adding more distant homologs to a multiple alignment and thus increasing its diversity may eventually deteriorate the numerical profile constructed from this alignment. Here, we addressed the question whether such a diversity limit can be reached in the alignments of confident homologs found by PSI-BLAST, and we analyzed the dependence of the quality of the profile-profile comparison made by COMPASS on the sequence diversity within these alignments. RESULTS: Protein families that have a greater number of diverse confident homologs in the current sequence databases provide an increased quality of similarity detection in profile databases, but produce on average less accurate profile-profile alignments with their remote relatives. This lower alignment accuracy cannot be improved when the most distant members of these families are excluded from their profiles. On the contrary, the presence of more diverse members results in more accurate alignments. For families with a high diversity of confident homologs, the lower quality of profile alignments with their remote relatives seems to be an attribute of these families or their alignments, rather than to be caused by the large number of diverse sequences itself. Our results suggest that at any level of profile diversity, one should include in the multiple alignment as many confident sequence homologs as possible in order to produce the most accurate results. PMID- 14751997 TI - ESPD: a pattern detection model underlying gene expression profiles. AB - MOTIVATION: DNA arrays permit rapid, large-scale screening for patterns of gene expression and simultaneously yield the expression levels of thousands of genes for samples. The number of samples is usually limited, and such datasets are very sparse in high-dimensional gene space. Furthermore, most of the genes collected may not necessarily be of interest and uncertainty about which genes are relevant makes it difficult to construct an informative gene space. Unsupervised empirical sample pattern discovery and informative genes identification of such sparse high dimensional datasets present interesting but challenging problems. RESULTS: A new model called empirical sample pattern detection (ESPD) is proposed to delineate pattern quality with informative genes. By integrating statistical metrics, data mining and machine learning techniques, this model dynamically measures and manipulates the relationship between samples and genes while conducting an iterative detection of informative space and the empirical pattern. The performance of the proposed method with various array datasets is illustrated. PMID- 14751998 TI - Comparative analysis of algorithms for signal quantitation from oligonucleotide microarrays. AB - MOTIVATION: Recent years' exponential increase in DNA microarrays experiments has motivated the development of many signal quantitation (SQ) algorithms. These algorithms perform various transformations on the actual measurements aimed to enable researchers to compare readings of different genes quantitatively within one experiment and across separate experiments. However, it is relatively unclear whether there is a 'best' algorithm to quantitate microarray data. The ability to compare and assess such algorithms is crucial for any downstream analysis. In this work, we suggest a methodology for comparing different signal quantitation algorithms for gene expression data. Our aim is to enable researchers to compare the effect of different SQ algorithms on the specific dataset they are dealing with. We combine two kinds of tests to assess the effect of an SQ algorithm in terms of signal to noise ratio. To assess noise, we exploit redundancy within the experimental dataset to test the variability of a given SQ algorithm output. For the effect of the SQ on the signal we evaluate the overabundance of differentially expressed genes using various statistical significance tests. RESULTS: We demonstrate our analysis approach with three SQ algorithms for oligonucleotide microarrays. We compare the results of using the dChip software and the RMAExpress software to the ones obtained by using the standard Affymetrix MAS5 on a dataset containing pairs of repeated hybridizations. Our analysis suggests that dChip is more robust and stable than the MAS5 tools for about 60% of the genes while RMAExpress is able to achieve an even greater improvement in terms of signal to noise, for more than 95% of the genes. PMID- 14751999 TI - Algorithms for variable length Markov chain modeling. AB - We present a general purpose implementation of variable length Markov models. Contrary to fixed order Markov models, these models are not restricted to a predefined uniform depth. Rather, by examining the training data, a model is constructed that fits higher order Markov dependencies where such contexts exist, while using lower order Markov dependencies elsewhere. As both theoretical and experimental results show, these models are capable of capturing rich signals from a modest amount of training data, without the use of hidden states. AVAILABILITY: The source code is freely available at http://www.soe.ucsc.edu/~jill/src/ PMID- 14752000 TI - CAAT-Box, Contigs-Assembly and Annotation Tool-Box for genome sequencing projects. AB - MOTIVATION: Contigs-Assembly and Annotation Tool-Box (CAAT-Box) is a software package developed for the computational part of a genome project where the sequence is obtained by a shotgun strategy. CAAT-Box contains new tools to predict links between contigs by using similarity searches with other whole genome sequences. Most importantly, it allows annotation of a genome to commence during the finishing phase using a gene-oriented strategy. For this purpose, CAAT Box creates an Individual Protein file (IPF) for each ORF of an assembly. The nucleotide sequence reported in an IPF corresponds to the sequence of the ORF with 500 additional bases before the ORF and 200 bases after. For annotation, additional information like Blast results can be added or linked to the IPFs as well as automatic and/or manual annotations. When a new assembly is performed, CAAT-Box creates new IPFs according to the old IPF panel. CAAT-Box recognizes the modified IPFs which are the only ones used for a new automatic analysis after each assembly. Using this strategy, the user works with a group of IPFs independently of the closure phase progression. The IPFs are accessible by a web server and can therefore be modified and commented by different groups. RESULT: CAAT-Box was used to obtain and to annotate several complete genomes like Listeria monocytogenes or Streptococcus agalactiae. AVAILABILITY: The program may be obtained from the authors and is freely available to non-profit organisations. PMID- 14752001 TI - In silico analysis of complete bacterial genomes: PCR, AFLP-PCR and endonuclease restriction. AB - We have developed a website, www.in-silico.com, which runs a software program that performs three basic tasks in completely sequenced bacterial genomes by in silico analysis: PCR amplification, amplified fragment length polymorphism (AFLP PCR) and endonuclease restriction. For PCR, after selection of the genome and introduction of primers, fragment size, DNA sequence and corresponding open reading frame (ORF) identity of the resulting PCR product is computed. Plasmids of sequenced species may be included in the analysis. Theoretical AFLP-PCR analyzes similar parameters, and includes a suggestion tool providing a list of commercial restriction enzyme pairs yielding up to 50 amplicons in the selected genome. Endonuclease restriction analysis of complete genomes and plasmids calculates the number of restriction sites for endonucleases in a given genome. If the number of fragments is 50 or fewer, pulsed field gel electrophoresis image and restriction maps are illustrated. Other tools that have been included in this site are ORF search by name and DNA to protein translation as well as restriction digestion of user-defined DNA sequences. AVAILABILITY: This is a new molecular biology resource freely available over the Internet at http://www.in-silico.com PMID- 14752002 TI - Gene selection for oligonucleotide array: an approach using PM probe level data. AB - MOTIVATION: Analysis of oligonucleotide array data, especially to select genes of interest, is a highly challenging task because of the large volume of information and various experimental factors. Moreover, interaction effect (i.e. expression changes depend on probe effects) complicates the analysis because current methods often use an additive model to analyze data. We propose an approach to address these issues with the aim of producing a more reliable selection of differentially expressed genes. The approach uses the rank for normalization, employs the percentile-range to measure expression variation, and applies various filters to monitor expression changes. RESULTS: We compare our approach with MAS and Dchip models. A data set from an angiogenesis study is used for illustration. Results show that our approach performs better than other methods either in identification of the positive control gene or in PCR confirmatory tests. In addition, the invariant set of genes in our approach provides an efficient way for normalization. PMID- 14752003 TI - Optimizing substitution matrices by separating score distributions. AB - MOTIVATION: Homology search is one of the most fundamental tools in Bioinformatics. Typical alignment algorithms use substitution matrices and gap costs. Thus, the improvement of substitution matrices increases accuracy of homology searches. Generally, substitution matrices are derived from aligned sequences whose relationships are known, and gap costs are determined by trial and error. To discriminate relationships more clearly, we are encouraged to optimize the substitution matrices from statistical viewpoints using both positive and negative examples utilizing Bayesian decision theory. RESULTS: Using Cluster of Orthologous Group (COG) database, we optimized substitution matrices. The classification accuracy of the obtained matrix is better than that of conventional substitution matrices to COG database. It also achieves good performance in classifying with other databases. PMID- 14752004 TI - Approximate geodesic distances reveal biologically relevant structures in microarray data. AB - MOTIVATION: Genome-wide gene expression measurements, as currently determined by the microarray technology, can be represented mathematically as points in a high dimensional gene expression space. Genes interact with each other in regulatory networks, restricting the cellular gene expression profiles to a certain manifold, or surface, in gene expression space. To obtain knowledge about this manifold, various dimensionality reduction methods and distance metrics are used. For data points distributed on curved manifolds, a sensible distance measure would be the geodesic distance along the manifold. In this work, we examine whether an approximate geodesic distance measure captures biological similarities better than the traditionally used Euclidean distance. RESULTS: We computed approximate geodesic distances, determined by the Isomap algorithm, for one set of lymphoma and one set of lung cancer microarray samples. Compared with the ordinary Euclidean distance metric, this distance measure produced more instructive, biologically relevant, visualizations when applying multidimensional scaling. This suggests the Isomap algorithm as a promising tool for the interpretation of microarray data. Furthermore, the results demonstrate the benefit and importance of taking nonlinearities in gene expression data into account. PMID- 14752006 TI - Guidelines for hypertension: are quality-assurance measures on target? AB - Guideline committees recommend targets of treatment based on trial data on efficacy and effectiveness. Quality-assurance initiatives apply these parameters in the general practice setting. Therefore, targets must be feasible and achievable by the practicing physicians who are judged by these targets as goals for care. We evaluated 437 patients in the Rush University Hypertension Clinic using the Health Employer Data Information Set (HEDIS) measures for 2000 to assess goal achievement in a practice-based setting. We compared guideline achievement of uncomplicated hypertensive and diabetic subjects to standards dictated by HEDIS, the 6th Joint National Committee on the Prevention, Detection, Evaluation, and Treatment of High Blood Pressure (JNC VI), and the American Diabetic Association (ADA)/National Kidney Foundation (NKF). Overall, 276 (63%) patients achieved SBP goal, with 376 (86%) achieving DBP goal and 358 (59%) achieving both goals. However, in the 20% of patients who were diabetic, only 52% had a BP of <140 mm Hg and <90 mm Hg, whereas only 22% achieved the more stringent goals of JNC VI of <130 mm Hg systolic and <85 mm Hg diastolic and only 15% achieved the ADA/NKF goals of <130 mm Hg systolic and <80 mm Hg diastolic. Although goal was achievable in most uncomplicated hypertension, hypertension in diabetes was more difficult to control, despite being more likely to receive enhanced benefit from effective management. Goal-oriented strategy, especially in diabetic subjects, should be aggressively sought rather than relaxing goals to promote achievement. PMID- 14752007 TI - The Mus81 solution to resolution: generating meiotic crossovers without Holliday junctions. PMID- 14752008 TI - Roles of Bifocal, Homer, and F-actin in anchoring Oskar to the posterior cortex of Drosophila oocytes. AB - Transport, translation, and anchoring of osk mRNA and proteins are essential for posterior patterning of Drosophila embryos. Here we show that Homer and Bifocal act redundantly to promote posterior anchoring of the osk gene products. Disruption of actin microfilaments, which causes delocalization of Bifocal but not Homer from the oocyte cortex, severely disrupts anchoring of osk gene products only when Homer (not Bifocal) is absent. Our data suggest that two processes, one requiring Bifocal and an intact F-actin cytoskeleton and a second requiring Homer but independent of intact F-actin, may act redundantly to mediate posterior anchoring of the osk gene products. PMID- 14752009 TI - Acf1 confers unique activities to ACF/CHRAC and promotes the formation rather than disruption of chromatin in vivo. AB - Chromatin assembly is required for the duplication of chromosomes. ACF (ATP utilizing chromatin assembly and remodeling factor) catalyzes the ATP-dependent assembly of periodic nucleosome arrays in vitro, and consists of Acf1 and the ISWI ATPase. Acf1 and ISWI are also subunits of CHRAC (chromatin accessibility complex), whose biochemical activities are similar to those of ACF. Here we investigate the in vivo function of the Acf1 subunit of ACF/CHRAC in Drosophila. Although most Acf1 null animals die during the larval-pupal transition, Acf1 is not absolutely required for viability. The loss of Acf1 results in a decrease in the periodicity of nucleosome arrays as well as a shorter nucleosomal repeat length in bulk chromatin in embryos. Biochemical experiments with Acf1-deficient embryo extracts further indicate that ACF/CHRAC is a major chromatin assembly factor in Drosophila. The phenotypes of flies lacking Acf1 suggest that ACF/CHRAC promotes the formation of repressive chromatin. The acf1 gene is involved in the establishment and/or maintenance of transcriptional silencing in pericentric heterochromatin and in the chromatin-dependent repression by Polycomb group genes. Moreover, cells in animals lacking Acf1 exhibit an acceleration of progression through S phase, which is consistent with a decrease in chromatin mediated repression of DNA replication. In addition, acf1 genetically interacts with nap1, which encodes the NAP-1 nucleosome assembly protein. These findings collectively indicate that ACF/CHRAC functions in the assembly of periodic nucleosome arrays that contribute to the repression of genetic activity in the eukaryotic nucleus. PMID- 14752010 TI - Rad6 plays a role in transcriptional activation through ubiquitylation of histone H2B. AB - Covalent modifications of the histone N tails play important roles in eukaryotic gene expression. Histone acetylation, in particular, is required for the activation of a subset of eukaryotic genes through the targeted recruitment of histone acetyltransferases. We have reported that a histone C tail modification, ubiquitylation of H2B, is required for optimal expression of several inducible yeast genes, consistent with a role in transcriptional activation. H2B was shown to be ubiquitylated and then deubiquitylated at the GAL1 core promoter following galactose induction. We now show that the Rad6 protein, which catalyzes monoubiquitylation of H2B, is transiently associated with the GAL1 promoter upon gene activation, and that the period of its association temporally overlaps with the period of H2B ubiquitylation. Rad6 promoter association depends on the Gal4 activator and the Rad6-associated E3 ligase, Bre1, but is independent of the histone acetyltransferase, Gcn5. The SAGA complex, which contains a ubiquitin protease that targets H2B for deubiquitylation, is recruited to the GAL1 promoter in the absence of H2B ubiquitylation. The data suggest that Rad6 and SAGA function independently during galactose induction, and that the staged recruitment of these two factors to the GAL1 promoter regulates the ubiquitylation and deubiquitylation of H2B. We additionally show that both Rad6 and ubiquitylated H2B are absent from two regions of transcriptionally silent chromatin but present at genes that are actively transcribed. Thus, like histone H3 lysine 4 and lysine 79 methylation, two modifications that it regulates, Rad6 directed H2B ubiquitylation defines regions of active chromatin. PMID- 14752012 TI - Diagnosis of neonatal sepsis: a clinical and laboratory challenge. PMID- 14752013 TI - High-level multiplex genotyping of polymorphisms involved in folate or homocysteine metabolism by matrix-assisted laser desorption/ionization mass spectrometry. AB - BACKGROUND: Increased plasma total homocysteine (tHcy), a risk factor for cardiovascular disease, is related to genetic, environmental, and nutritional factors, in particular folate status. Future large epidemiologic studies of the genetic basis of hyperhomocysteinemia will require high-throughput assays for polymorphisms of genes related to folate and Hcy metabolism. METHOD: We developed a high-level multiplex genotyping method based on matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) for the detection of 12 polymorphisms in 8 genes involved in folate or Hcy metabolism. The assay includes methylenetetrahydrofolate reductase (MTHFR) 677C>T and 1298A>C, methionine synthase (MTR) 2756A>G, methionine synthase reductase (MTRR) 66A>G, cystathionine beta-synthase (CBS) 844ins68 and 699C>T, transcobalamin II (TCII) 776C>G and 67A>G, reduced folate carrier-1 (RFC1) 80G>A, paraoxonase-1 (PON1) 575A>G and 163T>A, and betaine homocysteine methyltransferase (BHMT) 742G>A. RESULTS: The failure rate of the assay was < or = 1.7% and was attributable to unsuccessful DNA purification, nanoliter dispensing, and spectrum calibration. Most errors were related to identification of heterozygotes as homozygotes. The mean error rate was 0.26%, and error rates differed for the various single-nucleotide polymorphisms. Identification of CBS 844ins68 was carried out by a semiquantitative approach. The throughput of the MALDI-TOF MS assay was 1152 genotypes within 20 min. CONCLUSIONS: This high-level multiplex method is able to genotype 12 polymorphisms involved in folate or Hcy metabolism. The method is rapid and reproducible and could facilitate large-scale studies of the genetic basis of hyperhomocysteinemia and associated pathologies. PMID- 14752011 TI - Splicing enhances translation in mammalian cells: an additional function of the exon junction complex. AB - In mammalian cells, spliced mRNAs yield greater quantities of protein per mRNA molecule than do otherwise identical mRNAs not made by splicing. This increased translational yield correlates with enhanced cytoplasmic polysome association of spliced mRNAs, and is attributable to deposition of exon junction complexes (EJCs). Translational stimulation can be replicated by tethering the EJC proteins Y14, Magoh, and RNPS1 or the nonsense-mediated decay (NMD) factors Upf1, Upf2, and Upf3b to an intronless reporter mRNA. Thus, in addition to its previously characterized role in NMD, the EJC also promotes mRNA polysome association. Furthermore, the ability to stimulate translation when bound inside an open reading frame appears to be a general feature of factors required for NMD. PMID- 14752014 TI - Transcutaneous measurement of bilirubin in newborns: comparison with an automated Jendrassik-Grof procedure and HPLC. PMID- 14752015 TI - Serum S100B in pregnancy-related hypertensive disorders: a case-control study. PMID- 14752016 TI - Determination of thiopurine methyltransferase activity in isolated human erythrocytes does not reflect putative in vivo enzyme inhibition by sulfasalazine. PMID- 14752017 TI - Enzyme assay for diagnosis of guanidinoacetate methyltransferase deficiency. PMID- 14752018 TI - Nanogen microelectronic chip for large-scale genotyping. PMID- 14752019 TI - Rapid quantification of DNase I activity in one-microliter serum samples. PMID- 14752020 TI - Identification of twelve polymorphisms in the endothelin-1 gene by use of fluorescently labeled oligonucleotides and PCR with restriction fragment polymorphism analysis. PMID- 14752021 TI - IgA antibodies against tissue transglutaminase in the diagnosis of celiac disease: concordance with intestinal biopsy in children and adults. PMID- 14752022 TI - Time course of B-type natriuretic peptide (BNP) and N-terminal proBNP changes in patients with decompensated heart failure. PMID- 14752023 TI - Incidental clostebol contamination in athletes after sexual intercourse. PMID- 14752024 TI - Evaluation of the comparability of commercial ghrelin assays. PMID- 14752025 TI - Beckman Coulter access creatine kinase MB assay. PMID- 14752027 TI - Essential role for Smad3 in regulating MCP-1 expression and vascular inflammation. AB - Transforming growth factor (TGF)-beta(1) is a pleiotropic growth factor with known inhibitory effects on immune cell activation. However, the specific mechanism(s) and in vivo significance of the effectors of TGF-beta(1) modulation in the context of vascular inflammation are not well characterized. The chemokine monocyte chemoattractant protein (MCP)-1 is critical for the recruitment of macrophages in inflammatory disease states. In this study, we provide definitive evidence that the ability of TGF-beta(1) to inhibit MCP-1 expression is mediated via its effector Smad3. Adenoviral overexpression of Smad3 potently repressed inducible expression of endogenous MCP-1. Conversely, TGF-beta(1) inhibition of cytokine-mediated induction of MCP-1 expression was completely blocked in Smad3 deficient macrophages. Consistent with this impaired response, cardiac allografts in Smad3-deficient mice developed accelerated intimal hyperplasia with increased infiltration of adventitial macrophages expressing MCP-1. Previous studies show that MCP-1 inducibility is regulated by an AP-1 complex composed of c-Jun/c-Fos heterodimers. We demonstrate that the inhibitory effect of Smad3 occurs via a novel antagonistic effect of Smad3 on AP-1 DNA-protein binding and activity. Thus, Smad3 plays an essential role in modulating vascular inflammation characteristic of transplant-associated arteriopathy, is important in regulating MCP-1 expression, and plays a critical role in the ability of TGF-beta(1) to repress stimuli from a major inflammatory signaling pathway. PMID- 14752026 TI - Induction of hyaluronic acid synthase 2 (HAS2) in human vascular smooth muscle cells by vasodilatory prostaglandins. AB - Hyaluronic acid (HA) is a prominent constituent of the extracellular matrix of atherosclerotic vascular lesions in humans known to modulate vascular smooth muscle phenotype. The regulation of HA synthesis by vasodilatory prostaglandins was analyzed in human arterial smooth muscle cells (SMCs). The prostacyclin analogue, iloprost (100 nmol/L), markedly increased pericellular formation of HA coats and HA secretion into the cell culture medium in human arterial SMCs (8.7+/ 1.6-fold). Expression of HA synthase 2 (HAS2) was determined by semiquantitative RT-PCR and found to be strongly upregulated at concentrations of iloprost between 1 and 100 nmol/L after 3 hours. Furthermore, endogenous cyclooxygenase-2 (COX2) activity was required for basal expression of HAS2 mRNA in SMCs in vitro. Total HA secretion in response to iloprost was markedly decreased by RNA interference (RNAi), specific for HAS2. In addition, siRNA targeting HAS2 strongly increased the spreading of human SMCs compared with mock-transfected cells. HAS2 mRNA levels were also stimulated by a selective prostacyclin receptor (IP) agonist, cicaprost (10 nmol/L), prostaglandin E(2) (10 nmol/L), and the EP(2) receptor agonist, butaprost (1 micromol/L). Induction of HAS2 mRNA and HA synthesis by prostaglandins was mimicked by stable cAMP analogues and forskolin. In human atherectomy specimens from the internal carotid artery, HA deposits and COX2 expression colocalized frequently. In addition, strong EP(2) receptor expression was detected in SMCs in HA-rich areas. Therefore, upregulation of HAS2 expression via EP(2) and IP receptors might contribute to the accumulation of HA during human atherosclerosis, thereby mediating proatherosclerotic functions of COX2. PMID- 14752029 TI - Restitution of action potential duration during sequential changes in diastolic intervals shows multimodal behavior. AB - Restitution of action potential duration (APD) is thought to be critical in activation instability. Although restitution is used to predict APD during sequential changes in diastolic interval (DI), currently used protocols to determine restitution do not use sequential changes in DI. We explored restitution using a new pacing protocol to change DI sequentially and independently of APD. Transmembrane potentials were recorded from right ventricular endocardial tissue isolated from six dogs. We used three patterns of DIs: oscillatory, to demonstrate differences in APDs depending on previous activation history; random, to minimize effects of previous activation history, each DI preceding an APD had an equal probability of being short or long; and linear, to compare restitution relationship obtained during sequential changes in DI with those obtained using currently used protocols; DIs mimicked those that resulted using currently used protocols, except that they changed in sequence. During oscillatory DIs, restitution showed bimodal trajectory similar to hysteresis. Decrease in APD during decreasing DIs was faster than increase in APD during increasing DIs. When effects of previous activation history were minimized, we observed that for a given DI there were multiple values of APD. Restitution relationship obtained during sequential changes in DI was shallower than those obtained using currently used protocols. Our results show that the new pacing protocol may permit direct evaluation of effects of memory on APD. Sequential and explicit control of DI suggests that use of a unimodal relationship to predict APD when DIs change in sequence may not be appropriate. PMID- 14752028 TI - Role of Nrf2 in the regulation of CD36 and stress protein expression in murine macrophages: activation by oxidatively modified LDL and 4-hydroxynonenal. AB - CD36 is an important scavenger receptor mediating uptake of oxidized low-density lipoproteins (oxLDLs) and plays a key role in foam cell formation and the pathogenesis of atherosclerosis. We report the first evidence that the transcription factor Nrf2 is expressed in vascular smooth muscle cells, and demonstrate that oxLDLs cause nuclear accumulation of Nrf2 in murine macrophages, resulting in the activation of genes encoding CD36 and the stress proteins A170, heme oxygenase-1 (HO-1), and peroxiredoxin I (Prx I). 4-Hydroxy-2-nonenal (HNE), derived from lipid peroxidation, was one of the most effective activators of Nrf2. Using Nrf2-deficient macrophages, we established that Nrf2 partially regulates CD36 expression in response to oxLDLs, HNE, or the electrophilic agent diethylmaleate. In murine aortic smooth muscle cells, expressing negligible levels of CD36, both moderately and highly oxidized LDL caused only limited Nrf2 translocation and negligible increases in A170, HO-1, and Prx I expression. However, treatment of smooth muscle cells with HNE significantly enhanced nuclear accumulation of Nrf2 and increased A170, HO-1, and Prx I protein levels. Because PPAR-gamma can be activated by oxLDLs and controls expression of CD36 in macrophages, our results implicate Nrf2 as a second important transcription factor involved in the induction of the scavenger receptor CD36 and antioxidant stress genes in atherosclerosis. PMID- 14752030 TI - Modulation of in vivo cardiac function by myocyte-specific nitric oxide synthase 3. AB - Nitric oxide (NO) functions principally as a diffusible paracrine effector. The exception is in cardiomyocytes where both NO synthases (NOS) and target proteins coexist, allowing NO to work in an autocrine/intracrine fashion. However, the most abundant myocyte isoform (NOS3) is far more expressed in vascular endothelium; thus, the in vivo contribution of myocyte-NOS3 remains less clear. The present study tested this role by transfecting whole hearts of NOS3-null (NOS3(-/-)) mice with adenovirus-expressing NOS3 coupled to a alpha-MHC promoter (AdV(NOS3)), comparing results to hearts transfected with marker-gene beta galactosidase (AdVbeta(gal)). Total myocardial NOS3 protein and activity were restored to near wild-type (WT) levels in NOS3(-/-)+AdV(NOS3) hearts, and NOS3 relocalized normally with caveolin-3. Ejection function by pressure-volume analysis was enhanced in NOS3(-/-)+AdVbeta(gal) over WT or NOS3(-/-)+AdV(NOS3). More prominently, isoproterenol (ISO)-stimulated systolic and diastolic function in WT was amplified in NOS3(-/-)+AdVbeta(gal), whereas NOS3(-/-)+AdV(NOS3) returned the response to control. ISO-activated systolic function was inhibited 85% by concomitant muscarinic stimulation (carbachol) in NOS3(-/-)+AdV(NOS3) but not NOS3(-/-)+AdVbeta(gal) hearts. Lastly, NOS3(-/-)+AdVbeta(gal) mice displayed enhanced inotropy and lusitropy over WT at slower heart rates but a blunted rate augmentation versus controls. A more positive rate response was restored in NOS3( /-)+AdV(NOS3) (P<0.001). Thus, myocyte autocrine/intracrine NOS3 regulation in vivo can underlie key roles in beta-adrenergic, muscarinic, and frequency dependent cardiac regulation. PMID- 14752031 TI - Selective suppression of endothelial cell apoptosis by the high molecular weight form of adiponectin. AB - Adiponectin is an adipocyte-derived, antiatherogenic protein that is present in serum as three isoforms. Total adiponectin levels are decreased in obese or diabetic humans or animal models. This study was designed to elucidate the relative isoform distribution of adiponectin in human disease states and identify the active form of adiponectin toward vascular endothelial cells. The percentage of high molecular weight form (HMW) per total adiponectin was significantly lower in patients with coronary artery disease than control subjects, whereas the hexamer form was similar and the trimer form was significantly higher. During weight reduction in obese subjects, the HMW form increased and the trimer and hexamer forms decreased. Recombinant adiponectin dose-dependently suppressed apoptosis and caspase-3 activity in human umbilical vein endothelial cells (HUVECs). Transduction with dominant-negative AMP-activated protein kinase (AMPK) abolished the suppressive effect of adiponectin on HUVECs. Gel filtration chromatography was used to separate the adiponectin isoforms, and the antiapoptotic effect toward HUVECs was only observed with the HMW form. These data suggest that HMW adiponectin specifically confers the vascular-protective activities of this adipocytokine. The full text of this article is available online at http://circres.ahajournals.org. PMID- 14752032 TI - p21Cip1 levels differentially regulate turnover of mature endothelial cells, endothelial progenitor cells, and in vivo neovascularization. AB - p21(Cip1) (p21) controls cell cycle progression and apoptosis in mature endothelial cells (ECs) and regulates size and cycling of the hematopoietic progenitor cell pool. Because circulating endothelial progenitor cells (EPCs) contribute to postnatal neovascularization in addition to mature ECs, we investigated the regulation of ECs and EPCs in p21-deficient mice. Mature aortic EC proliferation was increased in homozygous p21(-/-) and heterozygous p21(+/-) mice, in which p21 protein levels are reduced to one third of wild-type (WT). In contrast, apoptosis sensitivity was increased by 3.5-fold only in p21(-/-), but not in p21(+/-) mice. Consistently, in vivo apoptosis of ECs within areas of neovascularization was elevated in p21(-/-) but not in p21(+/-) mice. EPC numbers were elevated 2-fold in p21(-/-) mice compared with WT (P<0.001), and clonal expansion capacity of EPCs was increased from 25+/-4 (WT) to 57+/-8 colony forming units in p21(-/-) mice (P<0.005). EPC numbers and expansion were likewise increased in p21(+/-) mice. As the integrative endpoint, in vivo neovascularization reflecting all p21-affected parameters was increased over WT only in p21(+/-) (P<0.001), but not in p21(-/-) mice. In conclusion, reduced p21 protein levels of mice lacking one p21 allele are associated with increased proliferation of ECs and EPCs, whereas survival of ECs to apoptotic stimuli in vitro and in vivo is not impaired. Under these conditions, neovascularization was increased. In contrast, complete p21 deficiency did not result in an increased neovascularization despite increased mature EC and EPC proliferation. This may be due to the sensitization of ECs against apoptosis. PMID- 14752033 TI - Sarcoplasmic reticulum calcium content fluctuation is the key to cardiac alternans. AB - The aim of this work was to investigate whether beat-to-beat alternation in the amplitude of the systolic Ca(2+) transient (Ca(2+) alternans) is due to changes of sarcoplasmic reticulum (SR) Ca(2+) content, and if so, whether the alternans arises due to a change in the gain of the feedback controlling SR Ca(2+) content. We found that, in rat ventricular myocytes, stimulating with small (20 mV) depolarizing pulses produced alternans of the amplitude of the Ca(2+) transient. Confocal measurements showed that the larger transients resulted from propagation of Ca(2+) waves. SR Ca(2+) content (measured from caffeine-evoked membrane currents) alternated in phase with the alternans of Ca(2+) transient amplitude. After a large transient, if SR Ca(2+) content was elevated by brief exposure of the cell to a Na(+)-free solution, then the alternans was interrupted and the next transient was also large. This shows that changes of SR Ca(2+) content are sufficient to produce alternans. The dependence of Ca(2+) transient amplitude on SR content was steeper under alternating than under control conditions. During alternation, the Ca(2+) efflux from the cell was also a steeper function of SR Ca(2+) content than under control. We attribute these steeper relationships to the fact that the larger responses in alternans depend on wave propagation and that wave propagation is a steep function of SR Ca(2+) content. In conclusion, alternans of systolic Ca(2+) appears to depend on alternation of SR Ca(2+) content. This, in turn results from the steep dependence on SR Ca(2+) content of Ca(2+) release and therefore Ca(2+) efflux from the cell as a consequence of wave propagation. PMID- 14752035 TI - The prenatal maturity of the accessory olfactory bulb in pigs. AB - The morphological development of the accessory olfactory bulb of the fetal pig was studied by classical and histo-chemical methods, and the vomeronasal organ and nasal septum were studied histochemically. Specimens were obtained from an abattoir and their ages estimated from their crown-to-rump length. The accessory olfactory bulb was structurally mature in fetuses of crown-to-rump length 21-23 cm, by which time the lectin Lycopersicum esculentum agglutinin stained the same structures as in adults (in particular, the entire sensory epithelium of the vomeronasal organ, the vomeronasal nerves, and the nervous and glomerular layers of the accessory olfactory bulb). These results suggest that the vomeronasal system of the pig may, like that of vertebrates such as snakes, be functional at birth. PMID- 14752036 TI - Analysis of a human fungiform papillae cDNA library and identification of taste related genes. AB - Various genes related to early events in human gustation have recently been discovered, yet a thorough understanding of taste transduction is hampered by gaps in our knowledge of the signaling chain. As a first step toward gaining additional insight, the expression specificity of genes in human taste tissue needs to be determined. To this end, a fungiform papillae cDNA library has been generated and analyzed. For validation of the library, taste-related gene probes were used to detect known molecules. Subsequently, DNA sequence analysis was performed to identify further candidates. Of 987 clones sequenced, clustering results in 288 contigs. Comparison of these contigs with genomic databases reveals that 207 contigs (71.9%) match known genes, 16 (5.6%) match hypothetical genes, eight (2.8%) match repetitive sequences and 57 (19.8%) have no or low similarity to annotated genes. The results indicate that despite a high level of redundancy, this human fungiform cDNA library contains specific taste markers and is valuable for investigation of both known and novel taste-related genes. PMID- 14752037 TI - Metallic taste and retronasal smell. AB - A series of experiments investigated the nature of metallic taste reports and whether they can be attributed to the development of a retronasal smell. Two studies showed that the metallic sensation reports following oral stimulation with solutions of FeSO4 were reduced to baseline when the nose was occluded. No such reduction was seen for CuSO4 or ZnSO4, which were more bitter and astringent, respectively, and less metallic. A discrimination test based on weak but equi-intense levels of FeSO4 and CuSO4 showed that FeSO4 could be discriminated from water with the nose open but not when occluded, but that discrimination of CuSO4 from water was not impaired by nasal occlusion. A discrimination test demonstrated that the headspace over solutions of FeSO4 was not different from water, although some subjects could discriminate FeSO4 solutions from water in the mouth when the nose was occluded, perhaps by tactile or astringent cues. These results confirm that metallic taste reports following oral stimulation with FeSO4 are likely due to development of a retronasal smell, possibly following a lipid oxidation reaction in the mouth. However, metallic taste reports may arise from different mechanisms with copper and zinc salts. PMID- 14752038 TI - Alarm pheromones with different functions are released from different regions of the body surface of male rats. AB - Our previous study suggested that the alarm pheromones in rats could be divided into at least two functionally different categories: one evoking autonomic responses and the other evoking behavioral responses, and the present study was conducted to test this hypothesis. Four regions of the body surface, i.e. the whisker pad, neck, rump and perianal region, of an anesthetized male Wistar rat were electrically stimulated (donor) and, after removal of the donor, the recipient rat was introduced into the same box and its behavioral and autonomic changes were recorded. Electrical stimulation of the perianal region of anesthetized donor rats provoked the release of odor that subsequently augmented core body temperature in other awake male rats. By contrast, electrical stimulation of the whisker pad of anesthetized donor males provoked the release of odor that augmented sniffing, rearing and locomotor activity in other awake male subjects. These results suggest that the alarm pheromone released from the face modifies behavior and that from the anal area induces autonomic stress responses in recipients. PMID- 14752039 TI - The influence of training on chemosensory event-related potentials and interactions between the olfactory and trigeminal systems. AB - It is not possible to accurately predict the perceptual response to odorants and odorant mixtures without understanding patterns of suppression and facilitation that result from interactions between the olfactory and trigeminal systems. The current study extends previous findings by exploring the effect of intensive training on the interaction between these systems and also by using a different mixed chemosensory stimulus to examine whether the principles established in earlier studies generalize to different odorants. Stimuli were chosen so as to selectively activate the olfactory (H2S) and trigeminal (CO2) nerves. In addition, linalool was included as a stimulus that activated both systems. Thirty five participants (19 men, 16 women) rated the intensity of each stimulus when presented both alone and in binary mixtures (linalool + H2S, and linalool + CO2). Chemosensory event-related potentials were obtained from three recording positions. Analysis of intensity ratings showed that linalool was significantly less intense than the other stimuli when presented alone. In binary mixtures, H2S was strongly suppressed by linalool. One week of intensive odor training produced significant and specific reductions in the intensity of linalool and H2S, both alone and in their mixture. Training with a different odor (champignol) had no effect. Chemosensory event-related potential data confirmed previous findings showing changes in topographical distribution that reflected the degree of trigeminal activity. Binary mixtures generally produced larger amplitudes than single stimuli. Latencies clearly differentiated between the three single stimuli and the binary mixtures. Changes were observed in event-related potentials that reflected those obtained for intensity ratings in that they were observed for linalool and H2S in the linalool trained group only. The amplitude of the late 'endogenous' component (P3) was significantly decreased for these odors at frontal recording sites. In summary, strong and specific training effects were observed in intensity ratings for participants trained with the test odor (linalool), but not for those trained with a different odor. This was supported by a significant decrease of amplitudes of the event-related potentials at frontal recording sites following training with the test odor only PMID- 14752040 TI - Individual differences in perception of bitterness from capsaicin, piperine and zingerone. AB - It was recently shown that in some subjects capsaicin can evoke bitterness as well as burning and stinging, particularly in the circumvallate (CV) region of the tongue. Because perception of bitterness from capsaicin is characterized by large individual differences, the main goal of the present study was to learn whether people who taste capsaicin as bitter also report bitterness from structurally similar sensory irritants that are known to stimulate capsaicin sensitive neurons. The irritancy and taste of capsaicin and two of its most commonly studied congeners, piperine and zingerone, were measured in individuals who had been screened for visibility of, and reliable access to, the CV papillae. Approximately half of these individuals reported tasting bitterness from all three irritants when the stimuli were swabbed directly onto the CV papillae. Concentrations that produced similar levels of burning sensation across subjects also produced similar (though lower) levels of bitter taste. These results are consistent with the hypothesis that capsaicin and its congeners stimulate bitterness via a common sensory receptor that is distributed differentially among individuals. Additionally, bitter tasters rated gustatory qualities (but not burning and stinging) slightly but significantly higher than did bitter non tasters, which suggests that perception of capsaicin bitterness is associated with a higher overall taste responsiveness (but not chemesthetic responsiveness) in the CV region. PMID- 14752041 TI - Voltage-gated inward currents of morphologically identified cells of the frog taste disc. AB - We used the patch clamp technique to record from taste cells in vertical slices of the bullfrog (Rana catesbeiana) taste disc. Cell types were identified by staining with Lucifer yellow in a pipette after recording their electrophysiological properties. Cells could be divided into the following three groups: type Ib (wing) cells with sheet-like apical processes, type II (rod) cells with single thick rod-like apical processes and type III (rod) cells with thin rod-like apical processes. No dye-coupling was seen either between cells of the same type or between cells of different types. We focused on the voltage gated inward currents of the three types of cells. Type Ib and type II cells exhibited tetrodotoxin (TTX)-sensitive voltage-gated Na+ currents. Surprisingly, type III cells showed TTX-resistant voltage-gated Na+ currents and exhibited a lack of TTX-sensitive Na+ currents. TTX-resistant voltage-gated Na+ currents in taste cells are reported for the first time here. The time constant for the inactivating portion of the voltage-gated inward Na+ currents of type III cells was much larger than that of type Ib and type II cells. Therefore, slow inactivation of inward Na+ currents characterizes type III cells. Amplitudes of the maximum peak inward currents of type III cells were smaller than those of type Ib and type II cells. However, the density (pA/pF) of the maximum peak inward currents of type III cells was much higher than that of type Ib cells and close to that of type II cells. No evidence of the presence of voltage-gated Ca2+ channels in frog taste cells has been presented up to now. In this study, voltage gated Ba2+ currents were observed in type III cells but not in type Ib and type II cells when the bath solution was a standard Ba2+ solution containing 25 mM Ba2+. Voltage-gated Ba2+ currents were blocked by addition of 2 mM CoCl2 to the standard Ba2+ solution, suggesting that type III cells possess the voltage-gated Ca2+ channels and they do classical (calcium-influx) synaptic transmission. It appears that type III cells are taste receptor cells. PMID- 14752042 TI - Intrinsic nitric oxide regulates the taste response of the sugar receptor cell in the blowfly, Phormia regina. AB - The taste organ in insects is a hair-shaped taste sensory unit having four functionally differentiated contact chemoreceptor cells. In the blowfly, Phormia regina, cGMP has been suggested to be a second messenger for the sugar receptor cell. Generally, cGMP is produced by membranous or soluble guanylyl cyclase (sGC), which can be activated by nitric oxide (NO). In the present paper, we electrophysiologically showed that an NO scavenger, 2-phenyl-4,4,5,5 tetramethylimidazoline-3-oxide-1-oxyl (PTIO), an NO donor, 1-hydroxy-2-oxo-3-(N methyl-3-aminopropyl)-3-methyl-1-triazene (NOC 7) or an NO synthase (NOS) inhibitor, NG-nitro-L-arginine methyl ester (L-NAME) specifically affected the response in the sugar receptor cell, but not in other receptor cells. PTIO, when introduced into the receptor cells in a sensillum aided by sodium deoxycholate (DOC, pH 7.2), depressed the response of sugar receptor cells to sucrose but did not affect those of the salt or water receptor cells. NOC 7, given extracellularly, latently induced the response of sugar receptor cells; and L NAME, when introduced into the receptor cells, depressed the response of sugar receptor cells. The results clearly suggest that NO, which may be produced by intrinsic NOS in sugar receptor cells, participates in the transduction cascade of these cells in blowfly. PMID- 14752043 TI - Taurine action on mitral cell activity in the frog olfactory bulb in vivo. AB - Taurine (TAU) is a free amino acid that is particularly abundant in the olfactory bulb. In the frog, TAU is located in the terminations of the primary olfactory axons and in the granular cell layer. TAU action seems to be associated with gamma amino butyric acid (GABA), the main inhibitory neurotransmitter involved in the processing of the sensory signal. The present study was designed to assess the action of TAU in vivo during the olfactory network's stimulation by odors. It was performed by recording the single-unit activity of mitral cells, the main bulbar output neurons. TAU effects were tested on both their spontaneous and odor induced firing activity. Interactions between TAU and GABA were examined by analyzing TAU effects under the selective blocking action of GABAA or GABAB antagonists. TAU was found to suppress the spontaneous firing of mitral cells, mainly without altering their odor response properties. By testing GABA antagonists, we further show that TAU action is associated with GABAergic inhibitory mechanisms mainly via GABAB receptors. Thus, TAU action clearly reduces background activity in favor of the emergence of the odor-induced activity in the same manner as GABA action does via GABAB receptors. As a conclusion, we propose that, in the frog olfactory bulb, the joint actions of TAU and GABA may favor the processing of the primary sensory information by increasing the signal to noise ratio. PMID- 14752044 TI - Oligonucleotide-directed site-specific integration of high complexity libraries into ssDNA templates. AB - We present an approach that generates an oligomer-based library with minimal need for restriction site modification of sequences in the target vector. The technique has the advantage that it can be applied for generating peptide aptamer libraries at sites within proteins without the need for introducing flanking enzyme sites. As an example we present a phagemid retroviral shuttle vector that can be used to achieve stable expression of the library in mammalian cells for the purpose of screening for peptides with desired biological activity. PMID- 14752045 TI - Catalytic and DNA-binding properties of the human Ogg1 DNA N-glycosylase/AP lyase: biochemical exploration of H270, Q315 and F319, three amino acids of the 8 oxoguanine-binding pocket. AB - The human Ogg1 protein (hOgg1) is an antimutator DNA glycosylase/AP lyase that catalyzes the excision of 8-oxo-7,8-dihydroguanine (8-oxoG) and the incision of apurinic and apyrimidinic (AP) sites in DNA. In this study, we have investigated the functional role of H270, Q315 and F319, three amino acids that are located in the 8-oxoG-binding pocket of hOgg1. Wild-type and mutant hOgg1 proteins (H270A, H270R, H270L, Q315A and F319A) were purified to apparent homogeneity. The catalytic activities and the DNA-binding properties of the various hOgg1 mutants were compared to those of the wild-type. The results show that hOgg1 mutated at H270 (H270A and H270L) or F319 (F319A) exhibits greatly reduced (50- to 1000 fold) DNA glycosylase activity, whereas the AP lyase activity is only moderately affected (<4-fold). The affinity of the hOgg1 mutants (H270A, H270L and F319A) for 8-oxoG.C-containing DNA is also greatly reduced (>30-fold), whereas their affinity for THF.C-containing DNA is only moderately reduced (<7-fold). The results also show that hOgg1 mutated at Q315 (Q315A) exhibits catalytic and DNA binding properties similar to those of the wild-type. Therefore, H270 and F319 are essential to form the functional 8-oxoG-binding pocket, whereas Q315 is less crucial. In contrast, H270, Q315 and F319 are not required for efficient binding of THF.C and cleavage of AP sites. Finally, hOgg1 mutant proteins with a substitution of H270A or F319A are members of a new type of hOgg1 that is deficient in DNA glycosylase but proficient in AP lyase. PMID- 14752046 TI - On the interpretation of Raman spectra of 1-aminooxy-spermine/DNA complexes. AB - By FT-Raman spectroscopy, we have investigated the effect of 1-aminooxy-spermine (AOSPM) on aggregation and stability of calf-thymus DNA and selected oligonucleotide chains. AOSPM is able to mimic spermine in some macromolecular interactions, but is unable to substitute polyamines to maintain cell proliferation, suggesting pharmacological applications. Raman spectra of solutions containing AOSPM and either genomic DNA or two 15mer oligodeoxyribonucleotides, with GC or AT sequences, were recorded. Precipitation was observed for calf-thymus DNA, aggregated structures and appearance of several Z marker bands were observed for the 15mer GC sequence, and no macromolecular changes were detected for the 15mer AT sequence. Specific binding sites between the aminooxy group and the base residues were also evidenced. Assignment of the AOSPM Raman bands was supported on a normal mode calculation for the molecule NH(2)-O-CH(3), as a model. The theoretical results, in combination with the analysis of the Raman bands, demonstrated that the aminooxy group played a relevant role in the AOSPM-DNA interaction. Preferential binding by the major groove was evidenced in the absence of macromolecular changes. When either precipitation or aggregation occurred, the interaction involved both the major and minor grooves. The specific interaction between AT/GC base pairs and the aminooxy group has also been theoretically investigated. The biological relevance of this work is discussed. PMID- 14752047 TI - Sequence-specific transcriptional repression by an MBD2-interacting zinc finger protein MIZF. AB - MBD2 is a member of the methyl-CpG-binding protein family that plays an important role in methylated DNA silencing. We have recently identified a novel zinc finger protein, MIZF, as an MBD2-binding partner. To understand the physiological function of MIZF in MBD2-mediated gene silencing, we investigated the DNA-binding properties of MIZF and its potential target genes. Using a cyclic amplification and selection of targets technique, the consensus sequence CGGACGTT, which contains a conserved CGGAC core, was determined as sufficient for MIZF binding. Deletion of individual zinc fingers revealed that five of the seven zinc fingers are required for DNA binding. Reporter assays demonstrated that MIZF represses transcription from the promoter including this DNA sequence. A database search indicated that a variety of human genes, including Rb, contain this sequence in their promoter region. MIZF actually bound to its recognition sequence within the Rb promoter and repressed Rb transcription. These results suggest that MIZF, through its DNA-binding activity, acts as a sequence-specific transcriptional repressor likely involved in MBD2-mediated epigenetic gene silencing. PMID- 14752048 TI - Modification of de novo DNA methyltransferase 3a (Dnmt3a) by SUMO-1 modulates its interaction with histone deacetylases (HDACs) and its capacity to repress transcription. AB - The de novo DNA methyltransferase Dnmt3a is one of three mammalian DNA methyltransferases that has been shown to play crucial roles in embryonic development, genomic imprinting and transcriptional silencing. Despite its importance, very little is known about how the enzymatic activity and transcriptional repression functions of Dnmt3a are regulated. Here we show that Dnmt3a interacts with multiple components of the sumoylation machinery, namely the E2 sumo conjugating enzyme Ubc9 and the E3 sumo ligases PIAS1 and PIASxalpha, all of which are involved in conjugating the small ubiquitin-like modifier polypeptide, SUMO-1, to its target proteins. Dnmt3a is modified by SUMO-1 in vivo and in vitro and the region of Dnmt3a responsible for interaction maps to the N terminal regulatory domain. Functionally, sumoylation of Dnmt3a disrupts its ability to interact with histone deacetylases (HDAC1/2), but not with another interaction partner, Dnmt3b. Conditions that enhance the sumoylation of Dnmt3a in vivo abolish its capacity to repress transcription. These studies reveal a new level of regulation governing Dnmt3a whereby a post-translational modification can dramatically regulate its interaction with specific protein partners and alter its ability to repress transcription. PMID- 14752050 TI - Linker phosphoramidite reagents for the attachment of the first nucleoside to underivatized solid-phase supports. AB - New linker phosphoramidite reagents containing a cleavable 3'-ester linkage are used for attaching the first nucleoside to the surface of a solid- phase support. Inexpensive, underivatized amino supports, such as long chain alkylamine controlled-pore glass, can serve as universal supports. No modifications to phosphoramidite coupling conditions are required and, after synthesis, treatment with NH(4)OH releases the products with 3'-OH ends. No 3'-dephosphorylation is required. Phosphoramidite reagents containing a succinate and sulfonyl diethanol linkage between the nucleoside and phosphoramidite group are particularly advantageous and can be used to create both 3'-OH and 5'-phosphate ends on oligonucleotides. Reproducibility and quality of oligonucleotide synthesis is demonstrated for either column and 96-well plate formats on low-, medium- or high loading CPG supports. PMID- 14752049 TI - Characterization of the 5'-untranslated region of YB-1 mRNA and autoregulation of translation by YB-1 protein. AB - The eukaryotic Y-box binding protein YB-1 is involved in various biological processes, including DNA repair, cell proliferation and the regulation of transcription and translation. YB-1 protein is abundant and expressed ubiquitously in human cells, functioning in cell proliferation and transformation. Its concentration is thought to be highly regulated at both the levels of transcription and translation. Therefore, we investigated whether or not the 5'-UTR of YB-1 mRNA affects the translation of YB-1 protein, thus influencing expression levels. Luciferase mRNA ligated to the YB-1 mRNA 5'-UTR was used as a reporter construct. Ligation of the full-length YB-1 5'-UTR (331 bases) enhanced translation as assessed by in vitro and in vivo translation assays. Deletion constructs of the YB-1 5'-UTR also resulted in a higher efficiency of translation, especially in the region mapped to +197 to +331 from the major transcription start site. RNA gel shift assays revealed that the affinity of YB-1 for various 5'-UTR probe sequences was higher for the full length 5'-UTR than for deleted 5'-UTR sequences. An in vitro translation assay was used to demonstrate that recombinant YB-1 protein inhibited translation of the full-length 5'-UTR of YB-1 mRNA. Thus, our findings provide evidence for the autoregulation of YB-1 mRNA translation via the 5'-UTR. PMID- 14752051 TI - Functional analysis of subcellular localization and protein-protein interaction sequences in the essential DNA ligase I protein of fission yeast. AB - DNA ligase I (Lig I) has key roles in chromosomal DNA replication and repair in the eukaryotic cell nucleus. In the budding yeast Saccharomyces cerevisiae the Lig I enzyme Cdc9p is also required for mitochondrial DNA replication and repair. In this report, dual nuclear-mitochondrial localization is demonstrated to be a property of the essential Lig I enzyme Cdc17 from the distantly related fission yeast Schizosaccharomyces pombe. Expression of nuclear and mitochondrial forms of Cdc17 from separate genes shows that, whereas expression of either protein alone is insufficient to restore viability to cells lacking endogenous Cdc17, co expression restores full viability. In the nucleus, Lig I interacts with the sliding clamp proliferating cell nuclear antigen (PCNA) via a conserved PCNA interacting sequence motif known as a PIP box. Deletion of the PIP motif from the N-terminus of the nuclear form of Cdc17 fails to abolish Cdc17 function, indicating that PCNA binding by Cdc17 is not an absolute requirement for completion of S-phase. PMID- 14752052 TI - Nuclear MxA proteins form a complex with influenza virus NP and inhibit the transcription of the engineered influenza virus genome. AB - Mx proteins belong to the dynamin superfamily of high molecular weight GTPases and interfere with multiplication of a wide variety of viruses. Earlier studies show that nuclear mouse Mx1 and human MxA designed to be localized in the nucleus inhibit the transcription step of the influenza virus genome. Here we set a transient influenza virus transcription system using luciferase as a reporter gene and cells expressing the three RNA polymerase subunits, PB1, PB2 and PA, and NP. We used this reporter assay system and nuclear-localized MxA proteins to get clues for elucidating the anti-influenza virus activity of MxA. Nuclear-localized VP16-MxA and MxA-TAg NLS strongly interfered with the influenza virus transcription. Over-expression of PB2 led to a slight resumption of the transcription inhibition by nuclear MxA, whereas over-expression of PB1 and PA did not affect the MxA activity. Of interest is that the inhibitory activity of the nuclear MxA was markedly neutralized by over-expression of NP. An NP devoid of its C-terminal region, but containing the N-terminal RNA binding domain, also neutralized the VP16-MxA activity in a dose-dependent manner, whereas an NP lacking the N-terminal region did not affect the VP16-MxA activity. Further, not only VP16-MxA but also the wild-type MxA was found to interact with NP in influenza virus-infected cells. This indicates that the nuclear MxA suppresses the influenza virus transcription by interacting with not only PB2 but also NP. PMID- 14752053 TI - Orphan nuclear receptor small heterodimer partner, a novel corepressor for a basic helix-loop-helix transcription factor BETA2/neuroD. AB - Small heterodimer partner (SHP; NR0B2) is an atypical orphan nuclear receptor that lacks a conventional DNA binding domain (DBD) and represses the transcriptional activity of various nuclear receptors. In this study, we examined the novel cross talk between SHP and BETA2/NeuroD, a basic helix-loop-helix transcription factor. In vitro and in vivo protein interaction studies showed that SHP physically interacts with BETA2/NeuroD, but not its heterodimer partner E47. Moreover, confocal microscopic study and immunostaining results demonstrated that SHP colocalized with BETA2 in islets of mouse pancreas. SHP inhibited BETA2/NeuroD-dependent transactivation of an E-box reporter, whereas SHP was unable to repress the E47-mediated transactivation and the E-box mutant reporter activity. In addition, SHP repressed the BETA2-dependent activity of glucokinase and cyclin-dependent kinase inhibitor p21 gene promoters. Gel shift and in vitro protein competition assays indicated that SHP inhibits neither dimerization nor DNA binding of BETA2 and E47. Rather, SHP directly repressed BETA2 transcriptional activity and p300-enhanced BETA2/NeuroD transcriptional activity by inhibiting interaction between BETA2 and coactivator p300. We also showed that C-terminal repression domain within SHP is also required for BETA2 repression. However, inhibition of BETA2 activity was not observed by naturally occurring human SHP mutants that cannot interact with BETA2/NeuroD. Taken together, these results suggest that SHP acts as a novel corepressor for basic helix-loop-helix transcription factor BETA2/NeuroD by competing with coactivator p300 for binding to BETA2/NeuroD and by its direct transcriptional repression function. PMID- 14752054 TI - Genetic analysis of adenohypophysis formation in zebrafish. AB - The adenohypophysis consists of at least six different cell types, somatotropes, lactotropes, thyrotropes, melanotropes, corticotropes, and gonadotropes. In mouse, cloning of spontaneous mutations and gene targeting has revealed multiple genes required for different steps of adenohypophysis development. Here, we report the results of a systematic search for genes required for adenohypophysis formation and patterning in zebrafish. By screening F3 offspring of N-ethyl-N nitrosourea-mutagenized founder fish, we isolated eleven mutants with absent or reduced expression of GH, the product of somatotropes, but a normally developing hypothalamus. Of such mutants, eight were further analyzed and mapped. They define four genes essential for different steps of adenohypophysis development. Two of them, lia and pia, affect the entire adenohypophysis, whereas the other two are required for a subset of adenohypophyseal cell types only. The third gene is zebrafish pit1 and is required for lactotropes, thyrotropes, and somatotropes, similar to its mouse ortholog, whereas the fourth, aal, is required for corticotropes, melanotropes, thyrotropes, and somatotropes, but not lactotropes. In conclusion, the isolated zebrafish mutants confirm principles of adenohypophysis development revealed in mouse, thereby demonstrating the high degree of molecular and mechanistic conservation among the different vertebrate species. In addition, they point to thus far unknown features of adenohypophysis development, such as the existence of a new lineage of pituitary cells, which partially overlaps with the Pit1 lineage. Positional cloning of the lia, pia, and aal genes might reveal novel regulators of vertebrate pituitary development. PMID- 14752056 TI - Insulin regulates protein kinase CbetaII alternative splicing in multiple target tissues: development of a hormonally responsive heterologous minigene. AB - Cells respond to external signals like insulin to alter metabolic pathways in response to varying physiological environments. Insulin stimulates the protein kinase C beta (PKCbeta) isozymes and preferentially switches the expression to PKCbetaII isozyme, which is shown to have a crucial role in glucose uptake, cellular proliferation, and differentiation. We have developed an insulin responsive PKCbetaII heterologous minigene to identify cis-elements in vivo in eukaryotes by cloning the PKCbetaII exon and its flanking intronic sequences into the splicing vector pSPL3. The transfected minigene mimicked the endogenous insulin response of PKCbetaII alternative splicing in five distinct cell types, i.e. L6 skeletal muscle, 3T3-L1 pre-adipocytes, HepG2 human hepatoma cells, A10 vascular smooth muscle cells, and murine embryonic fibroblasts within 30 min of insulin stimulation. Sequential deletions of the flanking introns in the minigene demonstrated that insulin regulated elements within the 5'-intron flanking the PKCbetaII exon. Mutational studies indicated the SRp40 binding site promotes splice site selection. In these cases, splicing appears to be regulated by a phosphatidylinositol 3-kinase signaling pathway because LY294002 and wortmannin, its specific inhibitors, blocked exon inclusion. Cotransfection with constitutively active Akt2 kinase mimicked insulin action. Signal-dependent regulation of splicing by insulin is unique from tissue-specific and developmentally regulated mechanisms previously reported and serves as a prototype for studies of alternative splicing involving protein phosphorylation. PMID- 14752058 TI - O-sulfonation of serine and threonine: mass spectrometric detection and characterization of a new posttranslational modification in diverse proteins throughout the eukaryotes. AB - Protein sulfonation on serine and threonine residues is described for the first time. This post-translational modification is shown to occur in proteins isolated from organisms representing a broad span of eukaryote evolution, including the invertebrate mollusk Lymnaea stagnalis, the unicellular malaria parasite Plasmodium falciparum, and humans. Detection and structural characterization of this novel post-translational modification was carried out using liquid chromatography coupled to electrospray tandem mass spectrometry on proteins including a neuronal intermediate filament and a myosin light chain from the snail, a cathepsin-C-like enzyme from the parasite, and the cytoplasmic domain of the human orphan receptor tyrosine kinase Ror-2. These findings suggest that sulfonation of serine and threonine may be involved in multiple functions including protein assembly and signal transduction. PMID- 14752059 TI - The fenfluramine metabolite (+)-norfenfluramine is vasoactive. AB - The anorexigen (+)-fenfluramine was used for treatment of obesity until the association of use with valvular heart disease and primary pulmonary hypertension. (+)-Fenfluramine has been found in Chinese and Korean slimming pills. The hepatic metabolite of (+)-fenfluramine, (+)-norfenfluramine, has affinity for 5-hydroxytryptamine (5-HT)(2A) and 5-HT(2B) receptors. We tested the hypothesis that (+)-norfenfluramine contracts arterial smooth muscle in a 5-HT receptor-dependent manner and acts as a pressor in the conscious rat. Isometric contraction experiments showed that (+)-norfenfluramine (10 nM, 100 microM) but not (+)-fenfluramine nor the isomer (-)-norfenfluramine caused concentration dependent contraction in arteries [-log EC(50) (moles per liter), thoracic aorta = 5.77 +/- 0.09; renal artery = 6.29 +/- 0.02; mesenteric resistance artery = 5.70 +/- 0.06]. Contraction was dependent on the 5-HT(2A) receptor because ketanserin (10 nM) rightward shifted (+)-norfenfluramine response curves (aorta = 16-fold, renal artery = 26-fold, and resistance artery = >100-fold). Dependence on activation of 5-HT(2A) receptors and independence of (+)-norfenfluramine induced contraction from stimulation of alpha-adrenergic receptors and the sympathetic nervous system was validated by demonstrating 1) unchanged contraction to (+)-norfenfluramine in arteries from chemically denervated rats; 2) a minimal effect of the alpha(1)-adrenergic receptor antagonist prazosin (100 nM) on contraction; and 3) antagonism by [6-methyl-l-(1-methylethy)ergoline-8beta carboxylic acid 2-hydroxy-1 methylpropyl ester maleate] LY53857 [6-methyl-1-(1 methylethy)-ergoline-8beta-carboxylic acid 2-hydroxy-1 methylpropyl ester maleate], a 5-HT(2) receptor antagonist without alpha-receptor affinity. (+) Norfenfluramine (10-300 microg/kg i.v.) caused a dose-dependent increase in mean arterial blood pressure in conscious rats, the maximum of which could be virtually abolished by ketanserin (3 mg/kg i.v.) but not prazosin (0.2 mg/kg i.v.). Our findings demonstrate for the first time that (+)-norfenfluramine is vasoactive and has the potential to increase blood pressure. PMID- 14752057 TI - Acute regulation of translation initiation by gonadotropin-releasing hormone in the gonadotrope cell line LbetaT2. AB - The hypothalamic neuropeptide hormone GnRH is the central regulator of reproductive function. GnRH stimulates the synthesis and release of the gonadotropins LH and FSH by the gonadotropes of the anterior pituitary through activation of the G-protein-coupled GnRH receptor. In this study, we investigated the role of translational control of hormone synthesis by the GnRH receptor in the novel gonadotrope cell line LbetaT2. Using immunohistochemical and RIA studies with this model, we show that acute GnRH-induced synthesis and secretion of LH are dependent upon new protein synthesis but not new mRNA synthesis. We examined the response to GnRH and found that activation of cap-dependent translation occurs within 4 h. LHbeta promoter activity was also examined, and we found no increases in LHbeta promoter activity after 6 h of GnRH stimulation. Additionally, we show that increased phosphorylation of translation initiation proteins, 4E-binding protein 1, eukaryotic initiation factor 4E, and eukaryotic initiation factor 4G, occur in a dose- and time-dependent manner in response to GnRH stimulation. Quantitative luminescent image analysis of Western blots shows that 10 nm GnRH is sufficient to cause a maximal increase in factor phosphorylation, and maximal responses occur within 30 min of stimulation. Further, we demonstrate that the MAPK kinase inhibitor, PD 98059, abolishes the GnRH-mediated stimulation of a cap-dependent translation reporter. More specifically, we demonstrate that PD 98059 abolishes the GnRH-mediated stimulation of a downstream target of the ERK pathway, MAPK-interacting kinase. Based on these findings, we conclude that acute GnRH stimulation of LbetaT2 cells increases translation initiation through ERK signaling. This may contribute to the acute increases in LHbeta subunit production. PMID- 14752060 TI - Atypical beta-adrenoceptor subtypes mediate relaxations of rabbit corpus cavernosum. AB - This study was performed to characterize the beta-adrenoceptor population in rabbit isolated corpus cavernosum (RbCC) by using nonselective and selective beta adrenoceptor agonists and antagonists in functional assays. Metaproterenol, ritodrine, fenoterol, and 8-hydroxy-5-[(1R)-1-hydroxy-2-[N-[(1R)-2-(rho methoxyphenyl)-1-methylethyl]amino]ethyl]carbostyril (TA 2005) (3-100 nmol each) dose dependently relaxed the RbCC preparations. These relaxations were markedly reduced by N(omega)-nitro-L-arginine methyl ester (L-NAME; 10 microM) and 1H [1,2,4]-oxadiazolo-[4,3,-a]quinoxalin-1-one (ODQ) (10 microM), whereas the adenylyl cyclase inhibitor SQ 22,536 [9-(2-tetrahydrofuryl) adenine] (10 microM) had no effect. In contrast, neither L-NAME nor ODQ affected the isoproterenol induced RbCC relaxations, but SQ 22,536 abolished this response. Sildenafil (1 microM) significantly potentiated the relaxations induced by beta(2)-agonists without affecting the isoproterenol-evoked relaxations. Rolipram (10 microM) enhanced the relaxations elicited by isoproterenol but had no effect on those induced by the selective beta(2) agonists. Propranolol and (+/-)-1-[2,3-(dihydro 7-methyl-1H-inden-4-yl)oxy]-3-[(1-methylethyl)amino]-2-butanol hydrochloride (ICI 118,551) determined a rightward shift in the concentration-response curves to isoproterenol in a noncompetitive manner with a reduction of maximum response at the highest antagonist concentration, with the slope values significantly different from unity. Propranolol and ICI 118,551 had no effect on the relaxations elicited by fenoterol, TA 2005, metaproterenol, and ritodrine. Atenolol and 1-[2-((3-carbamoyl-4-hydroxy)phenoxy) ethylamino]-3-[4-(1-methyl-4 trifluoromethyl-2-imidazolyl)-phenoxy]-2-propanol methanesulfonate (CGP 20712A) (0.1-10 microM) failed to affect the relaxations induced by all tested beta adrenoceptor agonists. Our study revealed the existence of two atypical beta adrenoceptors in the rabbit erectile tissue. Isoproterenol relaxes the rabbit cavernosal tissue by activating atypical beta-adrenoceptors coupled to adenylyl cyclase pathway, whereas the selective beta(2)-adrenoceptor agonists relax the RbCC tissue through another atypical beta-adrenoceptor subtype coupled to nitric oxide release from the sinusoidal endothelium. PMID- 14752061 TI - Fast onset medications through thermally generated aerosols. AB - Smoking involves heating a drug to form a mixture of drug vapor and gaseous degradation products. These gases subsequently cool and condense into aerosol particles that are inhaled. Here, we demonstrate rapid and reliable systemic delivery of pure pharmaceutical compounds without degradation products through a related process that also involves inhalation of thermally generated aerosol. Drug is coated as a thin film on a metallic substrate and vaporized by heating the metal. The thin nature of the drug coating minimizes the length of time during which the drug is exposed to elevated temperatures, thereby preventing its thermal decomposition. The vaporized, gas-phase drug rapidly condenses and coagulates into micrometer-sized aerosol particles. For the commonly prescribed antimigraine drug rizatriptan, inhalation of these particles results in nearly instantaneous systemic drug action. PMID- 14752062 TI - Histamine H1 and H2 receptor gene and protein levels are differentially expressed in the hearts of rodents and humans. AB - Histamine is highly concentrated in the heart of animals and humans. Excessive release in pathophysiological conditions, such as immediate hypersensitivity and septic shock, causes cardiac dysfunction and arrhythmias. Previous pharmacological studies revealed that H(1) and H(2) receptors mediate these effects. Yet, an accurate estimate of the distribution and molecular characteristics of cardiac histamine receptors is missing. Recently, the genes encoding H(1) and H(2) receptors have been cloned, and the amino acid sequence and protein structure have been elucidated. Accordingly, we analyzed gene and protein expression levels of H(1) and H(2) receptors in atria and ventricles of guinea pig, rabbit, rat, and human hearts. With immunocytochemical techniques, we examined the regional expression of H(1) and H(2) receptor proteins in the sinoatrial and atrioventricular nodes and surrounding myocardium of the guinea pig heart. Northern and Western blot studies revealed that cardiac histamine H(1) and H(2) receptors are variably distributed among different mammalian species and different regions of the heart, whereas H(2) receptors are abundantly expressed in human atrial and ventricular myocardium. These findings agree with those of previous pharmacological studies, clearly demonstrating that the responses of the heart to histamine depend on the expression level of H(1) and H(2) receptors. The highly abundant expression of H(2) receptors in the human heart substantiates histamine arrhythmogenicity in various disease states. The new knowledge of a differential distribution of histamine receptor subtypes in the human heart will foster a better understanding of histamine roles in cardiovascular pathophysiology and may contribute to new therapeutic approaches to histamine induced cardiac dysfunctions. PMID- 14752063 TI - Bone marrow chemoprotection without compromise of chemotherapy efficacy in a rat brain tumor model. AB - Thiol chemoprotective agents can reduce chemotherapy side effects, but clinical use is limited due to concerns of impaired chemotherapeutic efficacy. We evaluated whether an optimized bone marrow chemoprotection regimen impaired the efficacy of enhanced chemotherapy against rat brain tumors. Nude rats with intracerebral human lung carcinoma xenografts were treated with carboplatin, melphalan, and etoposide phosphate delivered intra-arterially with osmotic blood brain barrier disruption (n = 8/group). Thiol chemoprotection was N-acetyl-L cysteine (1000 mg/kg) 60 min before chemotherapy and/or sodium thiosulfate (8 g/m(2)) 4 and 8 h after chemotherapy, when the blood-brain barrier is reestablished. Blood counts were obtained before treatment on day 3 and at sacrifice on day 9. N-acetylcysteine serum clearance half-life was 9 to 11 min. Pretreatment with N-acetylcysteine combined with delayed administration of sodium thiosulfate protected against toxicity toward total white cells, granulocytes, and platelets (P = 0.0016). Enhanced chemotherapy reduced intracerebral tumor volume to 4.3 +/- 1.0 mm(3) compared with 29.1 +/- 4.1 mm(3) in untreated animals (P < 0.0001). Tumor volume was 3.7 +/- 0.6 mm(3) in rats that received N acetylcysteine before and sodium thiosulfate after chemotherapy. The data indicate the efficacy of enhanced chemotherapy for rat brain tumors was not affected by thiol chemoprotection that provided excellent protection for hematological toxicity. Negative interactions of thiols with antitumor efficacy were avoided by temporal and spatial separation of chemoprotectants and chemotherapy. PMID- 14752064 TI - Inhibition of nitric oxide generation by 23,24-dihydrocucurbitacin D in mouse peritoneal macrophages. AB - Nitric oxide (NO) has various physiological functions. However, uncontrolled overproduction of NO can be toxic in many pathologic conditions involving inflammatory tissue damage. In the present study, we examined effects of 23,24 dihydrocucurbitacin D (DHCD) isolated from the root of Bryonia alba L. on macrophage NO generation. DHCD (<80 microM) effectively abolished NO generation from macrophages activated with lipopolysaccharide and interferon-gamma. DHCD decreased the levels of protein and mRNA for inducible NO synthase (iNOS). DHCD potently blocked nuclear factor-kappaB (NF-kappaB) activation, a process necessary for transcriptional activation of iNOS. These results suggested that DHCD inhibited NO generation by blocking NF-kappaB activation and iNOS gene transcription. Because NF-kappaB activation is necessary not only for NO generation but also for many inflammatory processes, DHCD and its derivatives could be developed as anti-inflammatory drugs. PMID- 14752065 TI - Highly active antiretroviral therapy in AIDS-associated Kaposi's sarcoma: implications for the design of therapeutic trials in patients with advanced, symptomatic Kaposi's sarcoma. PMID- 14752066 TI - Low-dose thalidomide ameliorates cytopenias and splenomegaly in myelofibrosis with myeloid metaplasia: a phase II trial. AB - PURPOSE: A phase II dose-escalation trial was conducted to ascertain low-dose thalidomide safety and response in patients with advanced myelofibrosis with myeloid metaplasia (MMM). PATIENTS AND METHODS: Thalidomide was administered together with current therapy to 63 patients, starting at 50 mg daily and increasing to 400 mg as tolerated. RESULTS: Half of the patients sustained daily doses more than 100 mg and the drop-out rate was 51% at 6 months: the drop-out rate was lower in patients with high baseline fatigue score. At efficacy analysis, anemia was ameliorated in 22% of the patients and transfusions were eliminated in 39% of transfusion-dependent patients. Platelet count increased by 50 x 10(9)/L or more in 22% of patients with an initial count lower than 100 x 10(9)/L. Splenomegaly decreased by more than 50% of the initial size in 19% of patients. Reduction of an overall disease severity score occurred in 31% of patients and was associated with a significant reduction of fatigue. Disease severity amelioration was independently predicted by a high baseline myeloproliferative index (ie, large splenomegaly, thrombocytosis, or leukocytosis). CONCLUSION: Low-dose thalidomide displays an acceptable toxicity profile and provides an objective and subjective advantage to a relevant portion of MMM patients. PMID- 14752067 TI - Prognostic factors for survival in previously treated patients with metastatic renal cell carcinoma. AB - PURPOSE: To describe survival in previously treated patients with metastatic renal cell carcinoma (RCC) who are candidates for clinical trials of new agents as second-line therapy. PATIENTS AND METHODS: The relationship between pretreatment clinical features and survival was studied in 251 patients with advanced RCC treated during 29 consecutive clinical trials between 1975 and 2002. Clinical features were first examined in univariate analyses, and then a stepwise modeling approach based on Cox regression was used to form a multivariate model. RESULTS: Median survival for the 251 patients was 10.2 months and differed according to year of treatment, with patients treated after 1990 showing longer survival. In this group, the median overall survival time was 12.7 months. Because the purpose of this analysis was to establish prognostic factors for present-day clinical trial design, prognostic factor analysis was performed on these patients. Pretreatment features associated with a shorter survival in the multivariate analysis were low Karnofsky performance status, low hemoglobin level, and high corrected serum calcium. These were used as risk factors to categorize patients into three different groups. The median time to death in patients with zero risk factors was 22 months. The median survival in patients with one of these prognostic factors was 11.9 months. Patients with two or three risk factors had a median survival of 5.4 months. CONCLUSION: Treatment with novel agents during a clinical trial is indicated for patients with metastatic RCC after progression to cytokine treatment. Three prognostic factors for predicting survival were used to categorize patients into risk groups. These risk categories can be used in clinical trial design and interpretation. PMID- 14752068 TI - Relapse-free and overall survival in patients with pathologic stage II nonseminomatous germ cell cancer treated with etoposide and cisplatin adjuvant chemotherapy. AB - PURPOSE: To assess the long-term relapse-free survival and overall survival of patients with stage II nonseminomatous germ cell tumor (NSGCT) who received two cycles of adjuvant etoposide and cisplatin (EP) after primary retroperitoneal lymph node dissection. PATIENTS AND METHODS: Eighty-seven patients with completely resected pathologic stage II NSGCT were treated with adjuvant EP chemotherapy. Adjuvant EP consisted of two cycles of etoposide (100 mg/m(2)) plus cisplatin (20 mg/m(2)) per day, administered days 1 to 5 at a 21-day interval. RESULTS: Ten patients (11%) had pN1 disease, 73 (84%) had pN2 disease, and four (5%) had pN3 disease. Eighty-six patients received two cycles of EP, and one patient received an additional two cycles of EP after a transient marker increase after his first cycle. Eighty-seven patients are alive, and 86 patients (99%) remain relapse-free at a median follow-up of 8 years (range, 0.9 to 13.5 years). CONCLUSION: Two cycles of adjuvant EP is highly effective in preventing relapse in patients with pathologic stage II pN1 and pN2 NSGCT. An alternative treatment strategy is surveillance with full-course chemotherapy at relapse. Because there is a higher risk of relapse for patients with pN2 disease, these patients are offered adjuvant chemotherapy. PMID- 14752069 TI - Effect of duration of treatment on treatment outcome for patients with clear-cell sarcoma of the kidney: a report from the National Wilms' Tumor Study Group. AB - PURPOSE: To evaluate the effect of conventional and standard (ST) versus pulse intensive (PI) chemotherapy and short-duration versus long-duration chemotherapy on relapse-free survival (RFS) and overall survival rates of patients with clear cell sarcoma of the kidney (CCSK) entered onto the National Wilms' Tumor Study (NWTS)-4. PATIENTS AND METHODS: The 5-year and 8-year RFS rates were determined for patients with CCSK treated on the NWTS-4. After August 6, 1986, 40 previously untreated children younger than 16 years with CCSK were randomly assigned, after the completion of 6 months of chemotherapy, to discontinue (short) or continue 9 additional months (long) of treatment with chemotherapy regimens that included vincristine and either divided-dose (ST) courses (5 days) or single-dose (PI) treatment with dactinomycin and divided-dose (ST) courses (3 days) or single-dose (PI) treatment with doxorubicin. RESULTS: For patients with CCSK, the 5- and 8 year RFS rates were 65.2% and 60.6%, respectively, for patients randomly assigned to the short chemotherapy and 87.8% (both 5- and 8-year RFS) for patients randomly assigned to the long chemotherapy (P =.08). The overall survival rates for patients at 5 and 8 years were 95.5% and 85.9%, respectively, for the short chemotherapy and 87.5% (both 5- and 8-year overall survival) for the long chemotherapy (P =.99). In NWTS-4, the overall survival rates for patients with CCSK improved from NWTS-3 (83% v 66.9% at 8 years, respectively; P <.01). CONCLUSION: CCSK patients exhibit an improved RFS from a longer course of therapy when using vincristine, doxorubicin, and dactinomycin, but their long-term survival is unchanged compared with patients receiving 6 months of therapy. The overall survival rates for patients with CCSK have improved from NWTS-3. PMID- 14752070 TI - Combination of cytology, fluorescence in situ hybridization for aneuploidy, and reverse-transcriptase polymerase chain reaction for human mammaglobin/mammaglobin B expression improves diagnosis of malignant effusions. AB - PURPOSE: The identification of malignant cells in effusions by conventional cytology is hampered by its limited sensitivity. The aim of this study was to improve tumor cell detection in effusions by molecular approaches. MATERIALS AND METHODS: A total of 157 effusions from patients with tumors and 72 effusions from patients without a history or evidence of malignancy were included in this study. All effusion specimens were evaluated in parallel by cytology, fluorescence in situ hybridization (FISH) for aneuploidy, and reverse-transcriptase polymerase chain reaction (RT-PCR) for expression of human mammaglobin (hMAM) and mammaglobin B (hMAM-B). RESULTS: In effusions from patients with tumors, the sensitivities of tumor cell detection by cytology, FISH, and hMAM and hMAM-B detection were 46.2%, 53.3%, 36.4%, and 57.7%, respectively. The corresponding specificities were 94.4%, 97.0%, 87.1%, and 88.6%. Notably, a high percentage of effusions containing malignant cells were in fact transudates, indicating the necessity for molecular diagnostic work-up of transudates collected from patients with tumors. Dependent on the tumor type, the use of appropriate marker combinations improved tumor cell detection in effusions significantly. By combining all four diagnostic tests, a positive test result indicating the presence of malignancy was achieved in 81.1%, with a fairly good specificity of 70.1%. CONCLUSION: Molecular techniques are definitely useful to detect malignancy in cytologically negative effusions. Tumor cell detection in effusions can be significantly improved by FISH and PCR techniques applying appropriate molecular markers. This finding should help to improve tumor staging, prognostic assessment, and treatment monitoring. PMID- 14752072 TI - Surveillance of duodenal adenomas in familial adenomatous polyposis reveals high cumulative risk of advanced disease. AB - PURPOSE: The development of high-grade dysplasia (HGD) on duodenal or jejunal adenomas and of late-stage (stage IV) duodenal polyposis are major clinical events for familial adenomatous polyposis (FAP) patients. Our aim was to determine their respective frequency, risk factors, and cumulative risk. PATIENTS AND METHODS: A prospective, optimized, endoscopic surveillance protocol was applied to 58 FAP patients in a university hospital. The number, size, and histology of duodenojejunal polyps were assessed, and the Spigelman's score was calculated at each endoscopy. Cox regression and linear regression analysis were used to determine risk factors for HGD development and the cumulative risk of stage IV duodenal polyposis, respectively. RESULTS: During a median (+/- standard deviation) follow-up of 47.9 +/- 15.6 months, 35 patients with at least two consecutive examinations had 107 duodenojejunal examinations. The Spigelman's score increased in 21 patients (60.0%), and HGD developed in 12 patients (34.2%). High initial Spigelman's score (> 7 points), but not age or APC mutation site, was a risk factor for HGD development. Estimated cumulative risk of developing stage IV duodenal polyposis was of 42.9% at age 60 (95% CI, 35.7% to 50.0%) and 50.0% at age 70 (95% CI, 42.9% to 57.1%). CONCLUSION: This prospective series shows a higher duodenal polyposis progression rate and cumulative risk of late stage (stage IV) duodenal polyposis in FAP patients compared with previous series. These results suggest that current modalities for surveillance and management of these patients need revision. PMID- 14752071 TI - Efficacy of oral adjuvant therapy after resection of colorectal cancer: 5-year results from three randomized trials. AB - PURPOSE: Adjuvant therapy of colorectal cancer with oral fluorinated pyrimidines is attractive because of its ease of administration and good tolerability. The purpose of this meta-analysis is to assess the survival and disease-free survival benefits of treating patients after surgical resection of a primary colorectal tumor with oral fluoropyrimidines for 1 year. PATIENTS AND METHODS: This meta analysis was performed on individual data from three randomized trials conducted by the Japanese Foundation for Multidisciplinary Treatment for Cancer involving a total of 5,233 patients with stages I to III colorectal cancer. RESULTS: The overall hazard ratio in favor of oral therapy was 0.89 for survival (95% CI, 0.80 to 0.99; P =.04), and 0.85 for disease-free survival (95% CI, 0.77 to 0.93; P <.001). Thus oral therapy reduced the risk of death by 11% and the risk of recurrence or death by 15%. There was no significant heterogeneity between trials, nor did the benefit of oral therapy depend on tumor stage (I, II, or III), tumor site (rectum or colon), patient age, or patient sex. CONCLUSION: Oral fluoropyrimidines improve disease-free survival and survival of patients after resection of early-stage colorectal cancer. These observations support the use of these agents alone after resection of early-stage disease, as well as further testing of oral agents in combination with new drugs that have recently shown antitumor activity in advanced colorectal cancer. PMID- 14752073 TI - Urinary VEGF and MMP levels as predictive markers of 1-year progression-free survival in cancer patients treated with radiation therapy: a longitudinal study of protein kinetics throughout tumor progression and therapy. AB - PURPOSE: To determine the predictive value of urinary levels of two angiogenic factors, vascular endothelial growth factor (VEGF) and matrix metalloproteinase (MMPs), in a longitudinal study to determine their correlation with 1-year progression-free survival in patients with cancer. PATIENTS AND METHODS: VEGF and MMP levels were measured in the urine of 65 cancer patients at first evaluation, during therapy, and at follow-up (n = 242); normalized by creatinine levels; and compared with 16 healthy controls. The correlation of initial levels and trends of VEGF and MMPs with 1-year progression-free survival was assessed using two sample tests and stepwise logistic regression. RESULTS: Urinary VEGF levels at presentation were different between patients with local-regional cancer and normal controls, and between patients with metastatic prostate cancer and local regional disease (P =.04 and.01, respectively). Similar results were found with MMP measurement (P =.03 and.0001, respectively). Of those patients subsequently treated with radiation, VEGF levels at presentation between patients with no evidence of disease (NED) after radiation and those who had persistent or recurrent disease after radiotherapy were also different (P =.039). The comparison between angiogenic factor levels taken at least 1 month postradiotherapy and the last level taken during treatment was the strongest predictor of patient 1-year progression-free survival (P =.004). Similarly, the overall MMP trend was also significantly associated with 1-year progression-free survival, as was the individual MMP-2 trend (P =.004 and.001, respectively). Stepwise logistic regression revealed that the VEGF trend comparing postradiation levels with last level taken during treatment was an independent predictor of progression-free survival (P =.02). CONCLUSION: This small exploratory study suggests that the angiogenic urinary trends of VEGF and MMPs may be useful predictive markers for progression-free survival in cancer patients after the completion of radiotherapy. PMID- 14752074 TI - Effect of a cognitive behavioral intervention on reducing symptom severity during chemotherapy. AB - PURPOSE: To describe a randomized trial of a cognitive behavioral intervention on reducing symptom severity among patients diagnosed with solid tumors and undergoing a first course of chemotherapy and to determine whether the intervention had an additive or interactive effect on symptom severity in the presence of supportive care medications. PATIENTS AND METHODS: Patients (N = 237) were accrued from comprehensive and community cancer centers, interviewed, and randomly assigned to either the experimental intervention (n = 118) or conventional care (n = 119). A symptom severity index, based on summed severity scores across 15 symptoms, was the primary outcome. Each patient's site of cancer, stage at diagnosis, chemotherapy protocols, and use of supportive medications were learned from medical records. RESULTS: Groups were equivalent at baseline, and attrition by characteristics by group was not different. The proportion of patients not receiving chemotherapy at 10 and 20 weeks did not differ by group. At the 10- and 20-week observations, there was a significant interaction between the experimental group and baseline symptom severity. Patients in the experimental group who entered the trial with higher symptom severity reported significantly lower severity at 10 and 20 weeks. Controlling for chemotherapy treatment status at follow-up and supportive care medications did not alter the effect of the experimental intervention. CONCLUSION: Compared with conventional care alone, the experimental intervention was effective among patients who entered the trial with higher levels of symptom severity. Age, sex, site or stage of cancer, and supportive medications did not modify the effect of this cognitive behavioral intervention on symptom severity. PMID- 14752075 TI - Antioxidants and cancer therapy: a systematic review. AB - PURPOSE: Many patients with cancer take antioxidant nutritional supplements during cancer treatment to alleviate treatment toxicities and to improve long term outcomes, but little is known about the efficacy and safety of antioxidant use during cancer treatment. We reviewed English-language manuscripts published in the biomedical literature, reporting the results of observational studies of antioxidant status and cancer outcomes and of intervention trials of antioxidants among patients receiving chemotherapy with or without radiation for various malignancies. METHODS: We searched the Medline database and the bibliographies of the retrieved manuscripts, reviews, and books on antioxidants and cancer. The retrieved studies are grouped by study design, malignancy, and end points. RESULTS: More than 100 citations were retrieved; 52 met our criteria, 31 were observational studies, and 21 were intervention trials. The studies varied in study design, timing of observation/intervention, intervention protocol, malignancy, and anticancer regimen. CONCLUSION: These inconsistencies preclude a definitive conclusion as to the effect of chemotherapy on antioxidant status in patients undergoing anticancer therapy. However, our review suggests that total antioxidant status (measured by total radical antioxidant parameter) declines during cancer treatment. Adequately powered trials or observational studies among patients with a specific cancer diagnosis receiving a specific treatment regimen are needed to address patients' and physicians' concerns regarding these associations. PMID- 14752076 TI - Thymidylate synthase expression and prognosis in colorectal cancer: a systematic review and meta-analysis. AB - PURPOSE: A number of studies have investigated the relationship between thymidylate synthase (TS) expression and survival in colorectal cancer (CRC) patients. Although most have reported poorer overall and progression-free survival with high TS expression, estimates of the hazard ratio (HR) between studies differ wildly. To derive a more precise estimate of the prognostic significance of TS expression, we have reviewed published studies and carried out a meta-analysis. MATERIALS AND METHODS: Twenty studies stratifying overall survival and/or progression-free survival in CRC patients by TS expression status were eligible for analysis. The principal outcome measure was the HR. Data from these studies were pooled using standard meta-analysis techniques. RESULTS: Thirteen studies investigated outcome in a total of 887 cases with advanced CRC, and seven studies investigated outcome in a total of 2,610 patients with localized CRC. A number of methods were used both to assess TS expression and to assign TS status. Sample sizes varied greatly, small sample sizes being a feature of the advanced disease studies. The combined HR estimate for overall survival (OS) was 1.74 (95% CI, 1.34 to 2.26) and 1.35 (95% CI, 1.07 to 1.80) in the advanced and adjuvant settings, respectively, but there was evidence of heterogeneity and possible publication bias. CONCLUSION: Tumors expressing high levels of TS appeared to have a poorer OS compared with tumors expressing low levels. Additional studies with consistent methodology are needed to define the precise prognostic value of TS. PMID- 14752077 TI - Eligibility and outcomes reporting guidelines for clinical trials for patients in the state of a rising prostate-specific antigen: recommendations from the Prostate-Specific Antigen Working Group. AB - PURPOSE: To define methodology to show clinical benefit for patients in the state of a rising prostate-specific antigen (PSA). RESULTS: HYPOTHESIS: A clinical states framework was used to address the hypothesis that definitive phase III trials could not be conducted in this patient population. PATIENT POPULATION: The Group focused on men with systemic (nonlocalized) recurrence and a defined risk of developing clinically detectable metastases. Models to define systemic versus local recurrence, and risk of metastatic progression were discussed. INTERVENTION: Therapies that have shown favorable effects in more advanced clinical states; meaningful biologic surrogates of activity linked with efficacy in other tumor types; and/or effects on a target or pathway known to contribute to prostate cancer progression in this state can be considered for evaluation. OUTCOMES: An intervention-specific posttherapy PSA-based outcome definition that would justify further testing should be described at the outset. Reporting: Trial reports should include a table showing the number of patients who achieve a specific PSA-based outcome, the number who remain enrolled onto the trial, and the number who came off study at different time points. The term PSA response should be abandoned. TRIAL DESIGN: The phases of drug development for this state are optimizing dose and schedule, demonstration of a treatment effect, and clinical benefit. To move a drug forward should require a high bar that includes no rise in PSA in a defined proportion of patients for a specified period of time at a minimum. Agents that do not produce this effect can only be tested in combination. The preferred end point of clinical benefit is prostate cancer specific survival; the time to development of metastatic disease is an alternative. CONCLUSION: Methodology to show that an intervention alters the natural history of prostate cancer is described. At each stage of development, only agents with sufficient activity should be moved forward. PMID- 14752078 TI - Unusual lymphoma manifestations: case 1. Natural killer cell leukemia with dermal erythema and vascular epidermal growth factor production. PMID- 14752079 TI - Unusual lymphoma manifestations: case 2. Myocardial lymphoma presenting as atrial flutter. PMID- 14752080 TI - Unusual lymphoma manifestations: case 3. CD8+ T-cell prolymphocytic leukemia. PMID- 14752081 TI - The Day One Talk. PMID- 14752082 TI - Anaphylactoid reactions to isosulfan blue dye during breast cancer lymphatic mapping in patients given preoperative prophylaxis. PMID- 14752083 TI - HER2/neu expression and hormonal therapy in early breast cancer: can muddy waters become clear? PMID- 14752084 TI - Acute lymphoblastic leukemia-relapse study of the Berlin-Frankfurt-Munster Group (ALL-REZ BFM) experience: early treatment intensity makes the difference. PMID- 14752086 TI - Physician-patient communication in phase I cancer trials. PMID- 14752088 TI - Prospective, randomized, multicenter trial on the antiproliferative effect of lanreotide, interferon alfa, and their combination for therapy of metastatic neuroendocrine gastroenteropancreatic tumors. PMID- 14752089 TI - Is lanreotide and/or interferon alfa an adequate therapy for neuroendocrine tumors? PMID- 14752093 TI - Granzyme B encoded by the commonly occurring human RAH allele retains pro apoptotic activity. AB - A key function of human granzyme B (GrB) is to induce apoptosis of target cells in conjunction with perforin. The RAH allele is the first documented variant of the human GrB gene, occurs at a frequency of 25-30%, and encodes three amino acid substitutions (Q48R, P88A, and Y245H). It was initially reported that RAH GrB is incapable of inducing apoptosis, but here we show that it has essentially identical proteolytic and cytotoxic properties to wild type GrB. Recombinant RAH and wild type GrB cleave peptide substrates with similar kinetics, are both capable of cleaving Bid and procaspase 3, and are equally inhibited by proteinase inhibitor 9, an endogenous regulator of GrB. Furthermore, cytotoxic lymphocytes from RAH heterozygotes and homozygotes have no defect in target cell killing, and in vitro RAH GrB and wild type GrB kill cells equally well in the presence of perforin. We conclude that the RAH allele represents a neutral polymorphism in the GrB gene. PMID- 14752094 TI - Structural elements of instantaneous and slow gating in hyperpolarization activated cyclic nucleotide-gated channels. AB - Hyperpolarization-activated cyclic nucleotide-gated (HCN) subunits produce a slowly activating current in response to hyperpolarization (If) and an instantaneous voltage-independent current (Iinst) when expressed in Chinese hamster ovary (CHO) cells. Here we found that a mutation in the S4-S5 linker of HCN2 (Y331D) produced an additional mixed cationic instantaneous current. However, this current was inhibited by external Cs+ like If and unlike Iinst. Together with a concomitant reduction in If, the data suggest that the Y331D mutation disrupted channel closing placing the channel in a "If-like," and not an "Iinst-like," state. The "If-like" instantaneous current represented approximately 70% of total If over voltages ranging from +20 to -150 mV in high K+ solutions. If activated at more depolarized potentials and the activation curve was less steep, whereas deactivation was significantly slowed, consistent with the idea that the mutation inhibited channel closing. The data suggest that the mutation produced allosteric effects on the activation gate (S6 segment) and/or on voltage-sensing elements. We also found that decreases in the ratio of external K+/Na+ further disrupted channel closing in the mutant channel. Finally, our data suggest that the structures involved in producing Iinst are similar between the HCN1 and HCN2 isoforms and that excess HCN protein on the plasma membrane of CHO cells relative to native cells is not responsible for Iinst. The data are consistent with Iinst flowing through a "leaky" closed state but do not rule out flow through a second configuration of recombinant HCN channels or up regulated endogenous channels/subunits. PMID- 14752095 TI - Isoform-specific changes in scleral transforming growth factor-beta expression and the regulation of collagen synthesis during myopia progression. AB - The development of high myopia is associated with altered scleral extracellular matrix biochemistry. Previous studies highlight the importance of collagen turnover in this process, yet it is unclear which factors control scleral remodeling. This study used a mammalian model of myopia to investigate the capacity of TGF (transforming growth factor)-beta1, -beta2, and -beta3 to influence scleral remodeling in myopia. RT-PCR confirmed the presence of all mammalian TGF-beta isoforms in scleral tissue and scleral fibroblasts. Myopia was experimentally induced via monocular deprivation of pattern vision, and animals were allocated to two groups depending on the duration of treatment (1 or 5 days). Down-regulation of each isoform was apparent after only 1 day of myopia development (TGF-beta1, -32%; TGF-beta2, -27%; TGF-beta3, -42%). Whereas the decrease in TGF-beta1 and -beta3 expression was relatively constant between the two time points, differential down-regulation of TGF-beta2 was found between days 1 (-27%) and 5 (-50%). In vitro experiments, using primary scleral fibroblasts, demonstrated the capacity of all isoforms to increase collagen production in a dose-dependent manner. Changes in TGF-beta levels, which mimicked those during myopia induction, caused an approximately 15% reduction in collagen synthesis, which is qualitatively similar to those previously reported in vivo. These data represent the first demonstration of TGF-beta3 expression in the sclera and implicate all three TGF-beta isoforms in the control of scleral remodeling during myopia development. In addition, the early alterations in TGF-beta expression levels may reflect a role for these cytokines in mediating the retinoscleral signal that controls myopic eye growth. PMID- 14752096 TI - AML1 is functionally regulated through p300-mediated acetylation on specific lysine residues. AB - AML1 (RUNX1) is one of the most frequently disrupted genes in human leukemias. AML1 encodes transcription factors, which play a pivotal role in hematopoietic differentiation, and their inappropriate expression is associated with leukemic transformation of hematopoietic cells. Previous studies demonstrated that the transcription cofactor p300 binds to the C-terminal region of AML1 and stimulates AML1-dependent transcription during myeloid cell differentiation. Here, we report that AML1 is specifically acetylated by p300 in vitro. Mutagenesis analyses reveal that p300 acetylates AML1 at the two conserved lysine residues (Lys-24 and Lys-43). AML1 is subject to acetylation at the same sites in vivo, and p300 mediated acetylation significantly augments the DNA binding activity of AML1. Disruption of these two lysines severely impairs DNA binding of AML1 and reduced the transcriptional activity and the transforming potential of AML1. Taken together, these data indicate that acetylation of AML1 through p300 is a critical manner of posttranslational modification and identify a novel mechanism for regulating the function of AML1. PMID- 14752097 TI - 1-Methyl-4-phenylpyridinium-induced apoptosis in cerebellar granule neurons is mediated by transferrin receptor iron-dependent depletion of tetrahydrobiopterin and neuronal nitric-oxide synthase-derived superoxide. AB - In this study, we investigated the molecular mechanisms of toxicity of 1-methyl-4 phenylpyridinium (MPP(+)), an ultimate toxic metabolite of a mitochondrial neurotoxin, 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine, that causes Parkinson like symptoms in experimental animals and humans. We used rat cerebellar granule neurons as a model cell system for investigating MPP(+) toxicity. Results show that MPP(+) treatment resulted in the generation of reactive oxygen species from inhibition of complex I of the mitochondrial respiratory chain, and inactivation of aconitase. This, in turn, stimulated transferrin receptor (TfR)-dependent iron signaling via activation of the iron-regulatory protein/iron-responsive element interaction. MPP(+) caused a time-dependent depletion of tetrahydrobiopterin (BH(4)) that was mediated by H(2)O(2) and transferrin iron. Depletion of BH(4) decreased the active, dimeric form of neuronal nitric-oxide synthase (nNOS). MPP(+)-mediated "uncoupling" of nNOS decreased *NO and increased superoxide formation. Pretreatment of cells with sepiapterin to promote BH(4) biosynthesis or cell-permeable iron chelator and TfR antibody to prevent iron-catalyzed BH(4) decomposition inhibited MPP(+) cytotoxicity. Preincubation of cerebellar granule neurons with nNOS inhibitor exacerbated MPP(+)-induced iron uptake, BH(4) depletion, proteasomal inactivation, and apoptosis. We conclude that MPP(+) dependent aconitase inactivation, Tf-iron uptake, and oxidant generation result in the depletion of intracellular BH(4), leading to the uncoupling of nNOS activity. This further exacerbates reactive oxygen species-mediated oxidative damage and apoptosis. Implications of these results in unraveling the molecular mechanisms of neurodegenerative diseases (Parkinson's and Alzheimer's disease) are discussed. PMID- 14752098 TI - Structures of isobutyryl-CoA dehydrogenase and enzyme-product complex: comparison with isovaleryl- and short-chain acyl-CoA dehydrogenases. AB - The acyl-CoA dehydrogenases are a family of mitochondrial flavoproteins involved in the catabolism of fatty and amino acids. Isobutyryl-CoA dehydrogenase (IBD) is involved in the catabolism of valine and catalyzes the conversion of isobutyryl CoA to methacrylyl-CoA. The crystal structure of IBD with and without substrate has been determined to 1.76-A resolution. The asymmetric unit contains a homotetramer with substrate/product bound in two monomers. The overall structure of IBD is similar to those of previously determined acyl-CoA dehydrogenases and consists of an NH2-terminal alpha-helical domain, a medial beta-strand domain and a C-terminal alpha-helical domain. The enzyme-bound ligand has been modeled in as the reaction product, methacrylyl-CoA. The location of Glu-376 with respect to the C-2-C-3 of the bound product and FAD confirms Glu-376 to be the catalytic base. IBD has a shorter and wider substrate-binding cavity relative to short chain acyl-CoA dehydrogenase, permitting the optimal binding of the isobutyryl CoA substrate. The dramatic lateral expansion of the binding cavity seen in isovaleryl-CoA dehydrogenase is not observed in IBD. The conserved tyrosine or phenylalanine that defines a side of the binding cavity in other acyl-CoA dehydrogenases is replaced by a leucine (Leu-375) in the current structure. Substrate binding changes the position of some residues lining the binding pocket as well as the position of the loop containing the catalytic glutamate and subsequent helix. Three clinical mutations have been modeled to the structure. The mutations do not affect substrate binding but instead appear to disrupt protein folding and/or stability. PMID- 14752099 TI - How O2 binds to heme: reasons for rapid binding and spin inversion. AB - We have used density functional methods to calculate fully relaxed potential energy curves of the seven lowest electronic states during the binding of O(2) to a realistic model of ferrous deoxyheme. Beyond a Fe-O distance of approximately 2.5 A, we find a broad crossing region with five electronic states within 15 kJ/mol. The almost parallel surfaces strongly facilitate spin inversion, which is necessary in the reaction of O(2) with heme (deoxyheme is a quintet and O(2) a triplet, whereas oxyheme is a singlet). Thus, despite a small spin-orbit coupling in heme, the transition probability approaches unity. Using reasonable parameters, we estimate a transition probability of 0.06-1, which is at least 15 times larger than for the nonbiological Fe-O(+) system. Spin crossing is anticipated between the singlet ground state of bound oxyheme, the triplet and septet dissociation states, and a quintet intermediate state. The fact that the quintet state is close in energy to the dissociation couple is of biological importance, because it explains how both spin states of O(2) may bind to heme, thereby increasing the overall efficiency of oxygen binding. The activation barrier is estimated to be <15 kJ/mol based on our results and Mossbauer experiments. Our results indicate that both the activation energy and the spin transition probability are tuned by the porphyrin as well as by the choice of the proximal heme ligand, which is a histidine in the globins. Together, they may accelerate O(2) binding to iron by approximately 10(11) compared with the Fe-O(+) system. A similar near degeneracy between spin states is observed in a ferric deoxyheme model with the histidine ligand hydrogen bonded to a carboxylate group, i.e. a model of heme peroxidases, which bind H(2)O(2) in this oxidation state. PMID- 14752100 TI - Calmodulin interacts with the third intracellular loop of the serotonin 5 hydroxytryptamine1A receptor at two distinct sites: putative role in receptor phosphorylation by protein kinase C. AB - The serotonin 5-HT(1A) receptor couples to heterotrimeric G proteins and intracellular second messengers, yet no studies have investigated the possible role of additional receptor-interacting proteins in 5-HT(1A) receptor signaling. We have found that the ubiquitous Ca(2+)-sensor calmodulin (CaM) co immunoprecipitates with the 5-HT(1A) receptor in Chinese hamster ovary fibroblasts. The human 5-HT(1A) receptor contains two putative CaM binding motifs, located in the N- and C-terminal juxtamembrane regions of the third intracellular loop of the receptor. Peptides encompassing both the N-terminal (i3N) and C-terminal (i3C) CaM-binding domains were tested for CaM binding. Using in vitro binding assays in combination with gel shift analysis, we demonstrated Ca(2+)-dependent formation of complexes between CaM and both peptides. We determined kinetic data using a combination of BIAcore surface plasmon resonance (SPR) and dansyl-CaM fluorescence. SPR analysis gave an apparent K(D) of approximately 110 nm for the i3N peptide and approximately 700 nm for the i3C peptide. Both peptides also caused characteristic shifts in the fluorescence emission spectrum of dansyl-CaM, with apparent affinities of 87 +/- 23 nm and 1.70 +/- 0.16 microm. We used bioluminescence resonance energy transfer to show that CaM interacts with the 5-HT(1A) receptor in living cells, representing the first in vivo evidence of a G protein-coupled receptor interacting with CaM. Finally, we showed that CaM binding and phosphorylation of the 5-HT(1A) receptor i3 loop peptides by protein kinase C are antagonistic in vitro, suggesting a possible role for CaM in the regulation of 5-HT(1A) receptor phosphorylation and desensitization. These data suggest that the 5-HT(1A) receptor contains high and moderate affinity CaM binding regions that may play important roles in receptor signaling and function. PMID- 14752102 TI - Chemical synthesis and single channel properties of tetrameric and pentameric TASPs (template-assembled synthetic proteins) derived from the transmembrane domain of HIV virus protein u (Vpu). AB - Vpu, an 81-residue membrane protein encoded by the genome of HIV-1, is involved in CD4 degradation and facilitates virion budding from infected cells. The latter activity requires an intact transmembrane (TM) domain; however, the mechanism remains unclear. Vpu forms ion channels, an activity linked to the TM domain and envisioned to arise by oligomerization. The precise number of Vpu monomers that structure the channel is not yet known. To address this issue, we have synthesized tetrameric and pentameric proteins consisting of a carrier template to which four or five peptides corresponding to the TM domain of Vpu are attached. Ketoxime-forming chemoselective ligation efficiently ligated four and five copies, respectively, of the linear transmembrane peptide that was solubilized by the addition of a cleavable polyethylene glycol-polyamide auxiliary to a template. Purified tetrameric and pentameric proteins, denoted as T(4)Vpu and T(5)Vpu, exhibit the predicted mass as determined by MS analysis and fold with a high helical content as evidenced by CD. Both T(4)Vpu and T(5)Vpu, after reconstitution in lipid bilayers, form discrete ion channels of distinct conductance and high propensity to be open. The most frequent openings have a single channel conductance of 42 +/- 5 pS for T(4)Vpu and 76 +/- 5 pS for T(5)Vpu in 0.5m KCl. These findings validate the notion that the channels formed by Vpu result from the self-assembly of monomers. We conclude that a five-helix bundle of the TM of Vpu may approximate the structural motif underlying the oligomeric state of the conductive channel. PMID- 14752101 TI - Cytoplasmic polyadenylation element (CPE)- and CPE-binding protein (CPEB) independent mechanisms regulate early class maternal mRNA translational activation in Xenopus oocytes. AB - Meiotic cell cycle progression during vertebrate oocyte maturation requires the correct temporal translation of maternal mRNAs encoding key regulatory proteins. The mechanism by which specific mRNAs are temporally activated is unknown, although both cytoplasmic polyadenylation elements (CPE) within the 3' untranslated region (3'-UTR) of mRNAs and the CPE-binding protein (CPEB) have been implicated. We report that in progesterone-stimulated Xenopus oocytes, the early cytoplasmic polyadenylation and translational activation of multiple maternal mRNAs occur in a CPE- and CPEB-independent manner. We demonstrate that polyadenylation response elements, originally identified in the 3'-UTR of the mRNA encoding the Mos proto-oncogene, direct CPE- and CPEB-independent polyadenylation of an early class of Xenopus maternal mRNAs. Our findings refute the hypothesis that CPE sequences alone account for the range of temporal inductions of maternal mRNAs observed during Xenopus oocyte maturation. Rather, our data indicate that the sequential action of distinct 3'-UTR-directed translational control mechanisms coordinates the complex temporal patterns and extent of protein synthesis during vertebrate meiotic cell cycle progression. PMID- 14752103 TI - A regulatory domain in the C-terminal extension of the yeast glycerol channel Fps1p. AB - The Saccharomyces cerevisiae gene FPS1 encodes an aquaglyceroporin of the major intrinsic protein (MIP) family. The main function of Fps1p seems to be the efflux of glycerol in the adaptation of the yeast cell to lower external osmolarity. Fps1p is an atypical member of the family, because the protein is much larger (669 amino acids) than most MIPs due to long hydrophilic extensions in both termini. We have shown previously that a short domain in the N-terminal extension of the protein is required for restricting glycerol transport through the channel (Tamas, M. J., Karlgren, S., Bill, R. M., Hedfalk, K., Allegri, L., Ferreira, M., Thevelein, J. M., Rydstrom, J., Mullins, J. G. L., and Hohmann, S. (2003) J. Biol. Chem. 278, 6337-6345). Deletion of the N-terminal domain results in an unregulated channel, loss of glycerol, and osmosensitivity. In this work we have investigated the role of the Fps1p C terminus (139 amino acids). A set of eight truncations has been constructed and tested in vivo in a yeast fps1Delta strain. We have performed growth tests, membrane localization following cell fractionation, and glycerol accumulation measurements as well as an investigation of the osmotic stress response. Our results show that the C-terminal extension is also involved in restricting transport through Fps1p. We have identified a sequence of 12 amino acids, residues 535-546, close to the sixth transmembrane domain. This element seems to be important for controlling Fps1p function. Similar to the N-terminal domain, the C-terminal domain is amphiphilic and has a potential to dip into the membrane. PMID- 14752104 TI - Leptin counteracts sodium butyrate-induced apoptosis in human colon cancer HT-29 cells via NF-kappaB signaling. AB - This study shows that leptin induced a rapid phosphorylation of p42/44 mitogen activated protein kinase, an enhancement of both NF-kappaB DNA binding and transcriptional activities, and a concentration-dependent increase of HT-29 cell proliferation. These effects are consistent with the presence of leptin receptors on cell membranes. The leptin induction of cell growth was associated with an increase of cell population in S and G2/M phase compared with control cells found in G0/G1 phase of the cell cycle. Moreover, cyclin D1 immunoreactivity was enhanced in leptin-treated HT-29 cells and this increase was essentially associated with cell population in G0/G1 phase. On the other hand, we observed that sodium butyrate inhibited cell proliferation by blocking HT-29 cells in G0/G1 phase of the cell cycle. Interestingly, at physiological concentration, leptin prevented sodium butyrate-induced morphological nucleus changes, DNA laddering and suppressed butyrate-induced cell cycle arrest. This anti-apoptotic effect of leptin was associated with HT-29 cell proliferation and activation NF kappaB pathways. However, the phosphorylation of p42/44 MAP kinase in response to leptin was reduced in butyrate-treated cells. These data demonstrated that leptin is a potent mitogenic factor for intestinal epithelial cells through the MAP kinase and NF-kappaB pathways. They also showed, for the first time, that leptin promotes colon cancer HT-29 cell survival upon butyrate challenge by counteracting the apoptotic programs initiated by this short chain fatty acid probably through the NF-kappaB pathways. Although further studies are required to unravel the precise mechanism, these data may have significance in the pathogenesis of colorectal cancer and ulcerative colitis diseases. PMID- 14752105 TI - Cell type-specific activation of intracellular transglutaminase 2 by oxidative stress or ultraviolet irradiation: implications of transglutaminase 2 in age related cataractogenesis. AB - Transglutaminase (TGase) 2 is a ubiquitously expressed enzyme that modifies proteins by cross-linking or polyamination. An aberrant activity of TGase 2 has implicated its possible roles in a variety of diseases including age-related cataracts. However, the molecular mechanism by which TGase 2 is activated has not been elucidated. In this report, we showed that oxidative stress or UV irradiation elevates in situ TGase 2 activity. Neither the expression level nor the in vitro activity of TGase 2 appeared to correlate with the observed elevation of in situ TGase 2 activity. Screening a number of cell lines revealed that the level of TGase 2 activation depends on the cell type and also the environmental stress, suggesting that unrecognized cellular factor(s) may specifically regulate in situ TGase 2 activity. Concomitantly, we observed that human lens epithelial cells (HLE-B3) exhibited about 3-fold increase in in situ TGase 2 activity in response to the stresses. The activated TGase 2 catalyzed the formation of water-insoluble dimers or polymers of alphaB-crystallin, betaB(2) crystallin, and vimentin in HLE-B3 cells, providing evidence that TGase 2 may play a role in cataractogenesis. Thus, our findings indicate that in situ TGase 2 activity must be evaluated instead of in vitro activity to study the regulation mechanism and function of TGase 2 in biological and pathological processes. PMID- 14752106 TI - A novel actin bundling/filopodium-forming domain conserved in insulin receptor tyrosine kinase substrate p53 and missing in metastasis protein. AB - Insulin receptor tyrosine kinase substrate p53 (IRSp53) has been identified as an SH3 domain-containing adaptor that links Rac1 with a Wiskott-Aldrich syndrome family verprolin-homologous protein 2 (WAVE2) to induce lamellipodia or Cdc42 with Mena to induce filopodia. The recruitment of these SH3-binding partners by IRSp53 is thought to be crucial for F-actin rearrangements. Here, we show that the N-terminal predicted helical stretch of 250 amino acids of IRSp53 is an evolutionarily conserved F-actin bundling domain involved in filopodium formation. Five proteins including IRSp53 and missing in metastasis (MIM) protein share this unique domain and are highly conserved in vertebrates. We named the conserved domain IRSp53/MIM homology domain (IMD). The IMD has domain relatives in invertebrates but does not show obvious homology to any known actin interacting proteins. The IMD alone, derived from either IRSp53 or MIM, induced filopodia in HeLa cells and the formation of tightly packed parallel F-actin bundles in vitro. These results suggest that IRSp53 and MIM belong to a novel actin bundling protein family. Furthermore, we found that filopodium-inducing IMD activity in the full-length IRSp53 was regulated by active Cdc42 and Rac1. The SH3 domain was not necessary for IMD-induced filopodium formation. Our results indicate that IRSp53, when activated by small GTPases, participates in F-actin reorganization not only in an SH3-dependent manner but also in a manner dependent on the activity of the IMD. PMID- 14752108 TI - Phosphatidic acid and phosphatidylserine have distinct structural and functional interactions with the nicotinic acetylcholine receptor. AB - Bilayers containing phosphatidylcholine (PC) and the anionic lipid phosphatidic acid (PA) are particularly effective at stabilizing the nicotinic acetylcholine receptor (nAChR) in a functional conformation that undergoes agonist-induced conformational change. The physical properties of PC membranes containing PA are also substantially altered upon incorporation of the nAChR. To test whether or not the negative charge of PA is responsible for this "bi-directional coupling," the nAChR was reconstituted into membranes composed of PC with varying levels of the net negatively charged lipid phosphatidylserine (PS). In contrast to PA, increasing levels of PS in PC membranes do not stabilize an increasing proportion of nAChRs in a functional resting conformation, nor do they slow nAChR peptide hydrogen exchange kinetics. Incorporation of the nAChR had little effect on the physical properties of the PC/PS membranes, as monitored by the gel-to-liquid crystal phase transition temperatures of the bilayers. These results show that a net negative charge alone is not sufficient to account for the unique interactions that occur between the nAChR and PC/PA membranes. Incorporation of the receptor into PC/PS membranes, however, did lead to an altered head group conformation of PS possibly by recruiting divalent cations to the membrane surface. The results show that the nAChR has complex and unique interactions with both PA and PS. The interactions between the nAChR and PS may be bridged by divalent cations, such as calcium. PMID- 14752109 TI - Mutant R1 proteins from Escherichia coli class Ia ribonucleotide reductase with altered responses to dATP inhibition. AB - Aerobic ribonucleotide reductase from Escherichia coli regulates its level of activity by binding of effectors to an allosteric site in R1, located to the proposed interaction area of the two proteins that comprise the class I enzyme. Activity is increased by ATP binding and decreased by dATP binding. To study the mechanism governing this regulation, we have constructed three R1 proteins with mutations at His-59 in the activity site and one R1 protein with a mutation at His-88 close to the activity site and compared their allosteric behavior to that of the wild type R1 protein. All mutant proteins retained about 70% of wild type enzymatic activity. We found that if residue His-59 was replaced with alanine or asparagine, the enzyme lost its normal response to the inhibitory effect of dATP, whereas the enzyme with a glutamine still managed to elicit a normal response. We saw a similar result if residue His-88, which is proposed to hydrogen-bond to His 59, was replaced with alanine. Nucleotide binding experiments ruled out the possibility that the effect is due to an inability of the mutant proteins to bind effector since little difference in binding constants was observed for wild type and mutant proteins. Instead, the interaction between proteins R1 and R2 was perturbed in the mutant proteins. We propose that His-59 is important in the allosteric effect triggered by dATP binding, that the conserved hydrogen bond between His-59 and His-88 is important for the communication of the allosteric effect, and that this effect is exerted on the R1/R2 interaction. PMID- 14752107 TI - DNA-dependent protein kinase and checkpoint kinase 2 synergistically activate a latent population of p53 upon DNA damage. AB - The role of the checkpoint kinase 2 (Chk2) as an upstream activator of p53 following DNA damage has been controversial. We have recently shown that Chk2 and the DNA-dependent protein kinase (DNA-PK) are both involved in DNA damage-induced apoptosis but not G(1) arrest in mouse embryo fibroblasts. Here we demonstrate that Chk2 is required to activate p53 in vitro as measured by its ability to bind its consensus DNA target sequence following DNA damage and is in fact the previously unidentified factor working synergistically with DNA-PK to activate p53. The gene mutated in ataxia telangiectasia is not involved in this p53 activation. Using wortmannin, serine 15 mutants of p53, DNA-PK null cells and Chk2 null cells, we demonstrate that DNA-PK and Chk2 act independently and sequentially on p53. Furthermore, the p53 target of these two kinases represents a latent (preexisting) population of p53. Taken together, the results from these studies are consistent with a model in which DNA damage causes an immediate and sequential modification of latent p53 by DNA-PK and Chk2, which under appropriate conditions can lead to apoptosis. PMID- 14752110 TI - Activation of heme-regulated eukaryotic initiation factor 2alpha kinase by nitric oxide is induced by the formation of a five-coordinate NO-heme complex: optical absorption, electron spin resonance, and resonance raman spectral studies. AB - Heme-regulated eukaryotic initiation factor 2alpha kinase (HRI) regulates the synthesis of hemoglobin in reticulocytes in response to heme availability. HRI contains a tightly bound heme at the N-terminal domain. Earlier reports show that nitric oxide (NO) regulates HRI catalysis. However, the mechanism of this process remains unclear. In the present study, we utilize in vitro kinase assays, optical absorption, electron spin resonance (ESR), and resonance Raman spectra of purified full-length HRI for the first time to elucidate the regulation mechanism of NO. HRI was activated via heme upon NO binding, and the Fe(II)-HRI(NO) complex displayed 5-fold greater eukaryotic initiation factor 2alpha kinase activity than the Fe(III)-HRI complex. The Fe(III)-HRI complex exhibited a Soret peak at 418 nm and a rhombic ESR signal with g values of 2.49, 2.28, and 1.87, suggesting coordination with Cys as an axial ligand. Interestingly, optical absorption, ESR, and resonance Raman spectra of the Fe(II)-NO complex were characteristic of five coordinate NO-heme. Spectral findings on the coordination structure of full length HRI were distinct from those obtained for the isolated N-terminal heme binding domain. Specifically, six-coordinate NO-Fe(II)-His was observed but not Cys-Fe(III) coordination. It is suggested that significant conformational change(s) in the protein induced by NO binding to the heme lead to HRI activation. We discuss the role of NO and heme in catalysis by HRI, focusing on heme-based sensor proteins. PMID- 14752111 TI - End-to-end template jumping by the reverse transcriptase encoded by the R2 retrotransposon. AB - The reverse transcriptase encoded by the non-long terminal repeat retrotransposon R2 has been shown to be able to jump from the 5'-end of one RNA template (the donor) to the 3'-end of a second RNA template (the acceptor) in the absence of preexisting sequence identity between the two templates. These jumps between RNA templates have similarity to the end-to-end template jumps described for the RNA directed RNA polymerases encoded by certain RNA viruses. Here we describe for the first time the mechanism by which such end-to-end template jumps can occur. Most template jumps by the R2 reverse transcriptase are brought about by the enzyme's ability to add nontemplated (overhanging) nucleotides to the cDNA when it reaches the end of the donor RNA. The enzyme then anneals these overhanging nucleotides to sequences at the 3'-end of the acceptor RNA. The annealing is most efficient if it involves the terminal nucleotide(s) of the acceptor RNA but can occur to sites at least 5 nucleotides from the 3'-end. These end-to-end jumps are similar to steps proposed to be part of the integration reaction of non-long terminal repeat retrotransposons and can explain chimeric integration products derived from multiple RNA templates. PMID- 14752112 TI - Absence of hormone-sensitive lipase inhibits obesity and adipogenesis in Lep ob/ob mice. AB - Hormone-sensitive lipase (HSL) plays a crucial role in the hydrolysis of triacylglycerol and cholesteryl ester in various tissues including adipose tissues. To explore the role of HSL in the metabolism of fat and carbohydrate, we have generated mice lacking both leptin and HSL (Lep(ob/ob)/HSL(-/-)) by cross breeding HSL(-/-) mice with genetically obese Lep(ob/ob) mice. Unexpectedly, Lep(ob/ob)/HSL(-/-) mice ate less food, gained less weight, and had lower adiposity than Lep(ob/ob)/HSL(+/+) mice. Lep(ob/ob)/HSL(-/-) mice had massive accumulation of preadipocytes in white adipose tissues with increased expression of preadipocyte-specific genes (CAAT/enhancer-binding protein beta and adipose differentiation-related protein) and decreased expression of genes characteristic of mature adipocytes (CCAAT/enhancer-binding protein alpha, peroxisome proliferator activator receptor gamma, and adipocyte determination and differentiation factor 1/sterol regulatory element-binding protein-1). Consistent with the reduced food intake, hypothalamic expression of neuropeptide Y and agouti-related peptide was decreased. Since HSL is expressed in hypothalamus, we speculate that defective generation of free fatty acids in the hypothalamus due to the absence of HSL mediates the altered expression of these orexigenic neuropeptides. Thus, deficiency of both leptin and HSL has unmasked novel roles of HSL in adipogenesis as well as in feeding behavior. PMID- 14752113 TI - Seeding specificity in amyloid growth induced by heterologous fibrils. AB - Over residues 15-36, which comprise the H-bonded core of the amyloid fibrils it forms, the Alzheimer's disease plaque peptide amyloid beta (Abeta) possesses a very similar sequence to that of another short, amyloidogenic peptide, islet amyloid polypeptide (IAPP). Using elongation rates to quantify seeding efficiency, we inquired into the relationship between primary sequence similarity and seeding efficiency between Abeta-(1-40) and amyloid fibrils produced from IAPP as well as other proteins. In both a solution phase and a microtiter plate elongation assay, IAPP fibrils are poor seeds for Abeta-(1-40) elongation, exhibiting weight-normalized efficiencies of only 1-2% compared with Abeta-(1-40) fibrils. Amyloid fibrils of peptides with sequences completely unrelated to Abeta also exhibit poor to negligible seeding ability for Abeta elongation. Fibrils from a number of point mutants of Abeta-(1-40) exhibit intermediate seeding abilities for wild-type Abeta elongation, with differing efficiencies depending on whether or not the mutation is in the amyloid core region. The results suggest that amyloid fibrils from different proteins exhibit structural differences that control seeding efficiencies. Preliminary results also suggest that identical sequences can grow into different conformations of amyloid fibrils as detected by seeding efficiencies. The results have a number of implications for amyloid structure and biology. PMID- 14752114 TI - Tropomyosin exon 6b is troponin-specific and required for correct acto-myosin regulation. AB - The specificity of tropomyosin (Tm) exon 6b for interaction with and functioning of troponin (Tn) has been studied using recombinant fibroblast Tm isoforms 5a and 5b. These isoforms differ internally by exons 6a/6b and possess non-muscle exons 1b/9d at the termini, hence they lack the primary TnT(1)-tropomyosin interaction, allowing study of exon 6 exchange in isolation from this. Using kinetic techniques to measure regulation of myosin S1 binding to actin and fluorescently labeled Tm to directly measure Tn binding, we show that binding of Tn to both isoforms is similar (0.1-0.5 microm) and both produce well regulated systems. Calcium has little effect on Tn binding to the actin.Tm complex and both exons produce a 3-fold reduction in the S1 binding rate to actin.Tm.Tn in its absence. This confirms previous results that show exon 6 has little influence on Tn affinity to actin.Tm or its ability to fully inhibit the acto-myosin interaction. Thin filaments reconstituted with Tn and Tm5a or skeletal Tm (containing exon 6b) show nearly identical calcium dependence of acto-myosin regulation. However, Tm5b produces a dramatic increase in calcium sensitivity, shifting the activation mid point by almost an order of magnitude. This shows that exon 6 sequence and, hence, Tm structure in this region have a significant effect upon the calcium regulation of Tn. This finding supports evidence that familial hypertrophic cardiomyopathy mutations occurring adjacent to this region can effect calcium regulation. PMID- 14752115 TI - Differential amplification of intron-containing transcripts reveals long term potentiation-associated up-regulation of specific Pde10A phosphodiesterase splice variants. AB - We employed differential display of expressed mRNAs (Liang, P., and Pardee, A. B. (1992) Science 257, 967-971) to identify genes up-regulated after long term potentiation (LTP) induction in the hippocampus of awake adult rats. In situ hybridization confirmed the differential expression of five independently amplified clones representing two distinct transcripts, cl13/19/90 and cl95/96. Neither cl13/19/90 nor cl95/96 showed significant sequence homology to known transcripts (mRNA or expressed sequence tag) or to the mouse or human genome. However, comparison with the rat genome revealed that they are localized to a predicted intron of the phosphodiesterase Pde10A gene. cl13/19/90 and cl95/96 are likely to be part of the Pde10A primary transcript as, using reverse transcriptase-PCR, we could specifically amplify distinct introns of the Pde10A primary transcript, and in situ hybridization demonstrated that a subset of Pde10A splice variants are also up-regulated after LTP induction. These results indicate that amplification of a primary transcript can faithfully report gene activity and that differential display can be used to identify differential expression of RNA species other than mRNA. In transiently transfected Cos7 cells, Pde10A3 reduces the atrial natriuretic peptide-induced elevation in cGMP levels without affecting basal cGMP levels. This cellular function of LTP-associated Pde10A transcripts argues for a role of the cGMP/cGMP-dependent kinase pathway in long term synaptic plasticity. PMID- 14752116 TI - Human erythrocyte recycling of ascorbic acid: relative contributions from the ascorbate free radical and dehydroascorbic acid. AB - Recycling of ascorbic acid from its oxidized forms helps to maintain the vitamin in human erythrocytes. To determine the relative contributions of recycling from the ascorbate radical and dehydroascorbic acid, we studied erythrocytes exposed to a trans-membrane oxidant stress from ferricyanide. Ferricyanide was used both to induce oxidant stress across the cell membrane and to quantify ascorbate recycling. Erythrocytes reduced ferricyanide with generation of intracellular ascorbate radical, the concentrations of which saturated with increasing intracellular ascorbate and which were sustained over time in cells incubated with glucose. Ferricyanide also generated dehydroascorbic acid that accumulated in the cells and incubation medium to concentrations much higher than those of the radical, especially in the absence of glucose. Ferricyanide-stimulated ascorbate recycling from dehydroascorbic acid depended on intracellular GSH but was well maintained at the expense of intracellular ascorbate when GSH was severely depleted by diethylmaleate. This likely reflects continued radical reduction, which is not dependent on GSH. Erythrocyte hemolysates showed both NAD and NADPH-dependent ascorbate radical reduction. The latter was partially due to thioredoxin reductase. GSH-dependent dehydroascorbate reduction in hemolysates, which was both direct and enzyme-dependent, was greater than that of the radical reductase activity but of lower apparent affinity. Together, these results suggest an efficient two-tiered system in which high affinity reduction of the ascorbate radical is sufficient to remove low concentrations of the radical that might be encountered by cells not under oxidant stress, with back-up by a high capacity system for reducing dehydroascorbate under conditions of more severe oxidant stress. PMID- 14752118 TI - Association of the 16-kDa subunit c of vacuolar proton pump with the ileal Na+ dependent bile acid transporter: protein-protein interaction and intracellular trafficking. AB - The rat ileal apical sodium-dependent bile acid transporter (Asbt) transports conjugated bile acids in a Na+-dependent fashion and localizes specifically to the apical surface of ileal enterocytes. The mechanisms that target organic anion transporters to different domains of the ileal enterocyte plasma membrane have not been well defined. Previous studies (Sung, A.-Q., Arresa, M. A., Zeng, L., Swaby, I'K., Zhou, M. M., and Suchy, F. J. (2001) J. Biol. Chem. 276, 6825-6833) from our laboratory demonstrated that rat Asbt follows an apical sorting pathway that is brefeldin A-sensitive and insensitive to protein glycosylation, monensin treatment, and low temperature shift. Furthermore, a 14-mer signal sequence that adopts a beta-turn conformation is required for apical localization of rat Asbt. In this study, a vacuolar proton pump subunit (VPP-c, the 16-kDa subunit c of vacuolar H+-ATPase) has been identified as an interacting partner of Asbt by a bacterial two-hybrid screen. A direct protein-protein interaction between Asbt and VPP-c was confirmed in an in vitro pull-down assay and in an in vivo mammalian two-hybrid analysis. Indirect immunofluorescence confocal microscopy demonstrated that the Asbt and VPP-c colocalized in transfected COS-7 and MDCK cells. Moreover, bafilomycin A1 (a specific inhibitor of VPP) interrupted the colocalization of Asbt and VPP-c. A taurocholate influx assay and membrane biotinylation analysis showed that treatment with bafilomycin A1 resulted in a significant decrease in bile acid transport activity and the apical membrane localization of Asbt in transfected cells. Thus, these results suggest that the apical membrane localization of Asbt is mediated in part by the vacuolar proton pump associated apical sorting machinery. PMID- 14752117 TI - Structure of Kre2p/Mnt1p: a yeast alpha1,2-mannosyltransferase involved in mannoprotein biosynthesis. AB - Kre2p/Mnt1p is a Golgi alpha1,2-mannosyltransferase involved in the biosynthesis of Saccharomyces cerevisiae cell wall glycoproteins. The protein belongs to glycosyltransferase family 15, a member of which has been implicated in virulence of Candida albicans. We present the 2.0 A crystal structures of the catalytic domain of Kre2p/Mnt1p and its binary and ternary complexes with GDP/Mn(2+) and GDP/Mn(2+)/acceptor methyl-alpha-mannoside. The protein has a mixed alpha/beta fold similar to the glycosyltransferase-A (GT-A) fold. Although the GDP-mannose donor was used in the crystallization experiments and the GDP moiety is bound tightly to the active site, the mannose is not visible in the electron density. The manganese is coordinated by a modified DXD motif (EPD), with only the first glutamate involved in a direct interaction. The position of the donor mannose was modeled using the binary and ternary complexes of other GT-A enzymes. The C1" of the modeled donor mannose is within hydrogen-bonding distance of both the hydroxyl of Tyr(220) and the O2 of the acceptor mannose. The O2 of the acceptor mannose is also within hydrogen bond distance of the hydroxyl of Tyr(220). The structures, modeling, site-directed mutagenesis, and kinetic analysis suggest two possible catalytic mechanisms. Either a double-displacement mechanism with the hydroxyl of Tyr(220) as the potential nucleophile or alternatively, an S(N)i-like mechanism with Tyr(220) positioning the substrates for catalysis. The importance of Tyr(220) in both mechanisms is highlighted by a 3000-fold reduction in k(cat) in the Y220F mutant. PMID- 14752119 TI - Natural killer cell lytic activity and CD56(dim) and CD56(bright) cell distributions during and after intensive training. AB - The purpose of this study was to examine the impact of intensive training for competitive sports on natural killer (NK) cell lytic activity and subset distribution. Eight female college-level volleyball players undertook 1 mo of heavy preseason training. Volleyball drills were performed 5 h/day, 6 days/wk. Morning resting blood samples were collected before training (Pre), on the 10th day of training (During), 1 day before the end of training (End), and 1 wk after intensive training had ceased (Post). CD3(-)CD16(bright)CD56(dim) (CD56(dim) NK), CD3(-)CD16(dim/-)CD56(bright) NK (CD56(bright) NK), and CD3(+)CD16(-)CD56(dim) (CD56(dim) T) cells in peripheral blood were determined by flow cytometry. The circulating count of CD56(dim) NK cells (the predominant population, with a high cytotoxicity) did not change, nor did the counts for other leukocyte subsets. However, counts for CD56(bright) NK and CD56(dim) T cells (subsets with a lower cytotoxicity) increased significantly (P < 0.01) in response to the heavy training. Overall NK cell cytotoxicity decreased from Pre to End (P = 0.002), with a return to initial values at Post. Lytic units per NK cell followed a similar pattern (P = 0.008). Circulating levels of interleukin-6, interferon gamma, and tumor necrosis factor-alpha remained unchanged. These results suggest that heavy training can decrease total NK cell cytotoxicity as well as lytic units per NK cell. Such effects may reflect in part an increase in the proportion of circulating NK cells with a low cytotoxicity. PMID- 14752120 TI - Comparison of two plethysmography systems in assessment of forearm blood flow. AB - Venous occlusion plethysmography is widely used to assess forearm blood flow (FBF). We compared the established Hokanson system (HEC4) with a newly developed Filtrass 2001 system (F2001). The HEC4 uses mercury-in-Silastic strain gauges, whereas F2001 detects volume changes with a nonmercury linear displacement device. The aim of this study was to evaluate the new F2001 against the HEC4 in terms of repeatability and systematic bias. Ten subjects were studied on 4 separate days in random order using either the HEC4 on both arms, the F2001 on both arms, the HEC4 on the right arm with the F2001 on the left, or the F2001 on the right arm and the HEC4 on the left. Stroop's colored word conflict test and postocclusive hyperemia were used to increase FBF, and lower body negative pressure was used to lower FBF. Stroop's colored word conflict test and lower body negative pressure increased (24.6 +/- 1.5%, n = 240, P < 0.0001) and decreased (18.7 +/- 0.8%, n = 240, P < 0.0001) FBF, respectively. Postocclusive hyperemia after occlusion times of 5, 8, and 13 min substantially increased FBF by 390 +/- 86, 756 +/- 217, and 851 +/- 132%, respectively. Repeatability was not different between the devices (0.10 +/- 2.37 vs. -0.47 +/- 1.92 l/min, n = 125, P > 0.05), and there was no systematic bias. The F2001 is a newly developed plethysmography system that does not utilize mercury and is suitable for assessing changes of FBF in physiological studies. PMID- 14752121 TI - Inhibition of medullary raphe serotonergic neurons has age-dependent effects on the CO2 response in newborn piglets. AB - Medullary raphe serotonergic neurons are chemosensitive in culture and are situated adjacent to blood vessels in the brain stem. Selective lesioning of serotonergic raphe neurons decreases the ventilatory response to systemic CO2 in awake and sleeping adult rats. Abnormalities in the medullary serotonergic system, including the raphe, have been implicated in the sudden infant death syndrome (48). In this study, we ask whether serotonergic neurons in the medullary raphe and extra-raphe regions are involved in the CO2 response in unanesthetized newborn piglets, 3-16 days old. Whole body plethysmography was used to examine the ventilatory response to 5% CO2 before and during focal inhibition of serotonergic neurons by 8-hydroxy-2-di-n-propylaminotetralin (8-OH DPAT), a 5-HT1A receptor agonist. 8-OH-DPAT (10 or 30 mM in artificial cerebrospinal fluid) decreased the CO2 response in wakefulness in an age dependent manner, as revealed by a linear regression analysis that showed a significant negative correlation (P < 0.001) between the percent change in the CO2 response and piglet age. Younger piglets showed an exaggerated CO2 response. Control dialysis with artificial cerebrospinal fluid had no significant effect on the CO2 response. Additionally, 8-OH-DPAT increased blood pressure and decreased heart rate independent of age (P < 0.05). Finally, sleep cycling was disrupted by 8-OH-DPAT, such that piglets were awake more and asleep less (P < 0.05). Because of the fragmentary sleep data, it was not possible to examine the CO2 response in sleep. Inhibition of serotonergic medullary raphe and extra-raphe neurons decreases ventilatory CO2 sensitivity and alters cardiovascular variables and sleep cycling, which may contribute to the sudden infant death syndrome. PMID- 14752122 TI - Effect of prenatal nicotine exposure on biphasic hypoxic ventilatory response and protein kinase C expression in caudal brain stem of developing rats. AB - Current evidence suggests that maternal smoking is associated with decreased respiratory drive and blunted hypoxic ventilatory response (HVR) in the newborn. The effect of prenatal nicotine exposure on overall changes in HVR has been studied; however, there is limited data on the effect of nicotine exposure on each component of biphasic HVR. To examine this issue, 5-day timed-pregnant Sprague-Dawley rats underwent surgical implantation of an osmotic minipump containing either normal saline (Con) or a solution of nicotine tartrate (Nic) to continuously deliver free nicotine at 6 mg.kg of maternal weight(-1).day(-1). Rat pups at postnatal days 5, 10, 15, and 20 underwent hypoxic challenges with 10% O(2) for 20 min using whole body plethysmography. At postnatal day 5, Nic was associated with attenuation of peak HVR; peak minute ventilaton increased 44.0 +/ 6.8% (SE) from baseline in Nic pups, whereas that of Con pups increased 62.9 +/- 5.1% (P < 0.05). Nic pups also had a reduction in the magnitude of ventilatory roll-off; minute ventilation at 15 min decreased 7.3 +/- 7.1% in Nic pups compared with 27.3 +/- 4.0% in Con pups (P < 0.05). No significant difference in HVR was noted at postnatal days 10, 15, and 20. Hypercapnic response was similar at all ages. We further investigated the effect of prenatal nicotine exposure on PKC expression in the caudal brain stem (CB) of developing rats. At postnatal day 5, Nic was associated with increased expression of PKC-beta and PKC-delta in CB, whereas other PKC isoforms were not affected. It is concluded that prenatal nicotine exposure is associated with modulation of biphasic HVR and a selective increase in the expression of PKC-beta and PKC-delta within the CB of developing rats. PMID- 14752123 TI - Hypercholesterolemia inhibits L-type calcium current in coronary macro-, not microcirculation. AB - Hypercholesterolemia (HC) is a mary risk factor for the development of coronary heart disease. Coronary ion regulation, especially calcium, is thought to be important in coronary heart disease development; however, the influence of high dietary fat and cholesterol on coronary arterial smooth muscle (CASM) ion channels is unknown. The purpose of this study was to determine the effect of diet-induced HC on CASM voltage-gated calcium current (I(Ca)). Male miniature swine were fed a high-fat, high-cholesterol diet (40% kcal fat, 2% wt cholesterol) for 20-24 wk, resulting in elevated serum total and low-density lipoprotein cholesterol. Histochemistry indicated early atherosclerosis in large coronary arteries. CASM were isolated from the right coronary artery (>1.0 mm ID), small arteries ( approximately 200 microm), and large arterioles ( approximately 100 microm). I(Ca) was determined by whole cell voltage clamp. L type I(Ca) was reduced approximately 30% by HC compared with controls in the right coronary artery (-5.29 +/- 0.42 vs. -7.59 +/- 0.41 pA/pF) but not the microcirculation (small artery, -8.39 +/- 0.80 vs. -10.13 +/- 0.60; arterioles, 10.78 +/- 0.93 vs. -11.31 +/- 0.95 pA/pF). Voltage-dependent activation was unaffected by HC in both the macro- and microcirculation. L-type voltage-gated calcium channel (Ca(v)1.2) mRNA and membrane protein levels were unaffected by HC. Inhibition of I(Ca) by HC was reversed in vitro by the cholesterol scavenger methyl-beta-cyclodextrin and mimicked in control CASM by incubation with the cholesterol donor cholesterol:methyl-beta-cyclodextrin. These data indicate that CASM L-type I(Ca) is decreased in large coronary arteries in early stages of atherosclerosis, whereas I(Ca) in the microcirculation is unaffected. The inhibition of calcium channel activity in CASM of large coronary arteries is likely due to increases in membrane free cholesterol. PMID- 14752124 TI - Exercise training improves aortic endothelium-dependent vasorelaxation and determinants of nitric oxide bioavailability in spontaneously hypertensive rats. AB - The present study examined in vitro vasomotor function and expression of enzymes controlling nitric oxide (NO) bioavailability in thoracic aorta of adult male normotensive Wistar-Kyoto (WKY) and spontaneously hypertensive rats (SHR) that either remained sedentary (Sed) or performed 6 wk of moderate aerobic exercise training (Ex). Training efficacy was confirmed by elevated maximal activities of both citrate synthase (P = 0.0024) and beta-hydroxyacyl-CoA dehydrogenase (P = 0.0073) in the white gastrocnemius skeletal muscle of Ex vs. Sed rats. Systolic blood pressure was elevated in SHR vs. WKY (P < 0.0001) but was not affected by Ex. Despite enhanced endothelium-dependent relaxation to 10(-8) M ACh in SHR vs. WKY (P = 0.0061), maximal endothelium-dependent relaxation to 10(-4) M ACh was blunted in Sed SHR (48 +/- 12%) vs. Sed WKY (84 +/- 6%, P = 0.0067). Maximal endothelium-dependent relaxation to 10(-4) M ACh was completely restored in Ex SHR (93 +/- 9%) vs. Sed SHR (P = 0.0011). N(omega)-nitro-l-arginine abolished endothelium-dependent relaxation in all groups (P or =8% was at least 80%. CONCLUSIONS: Combined thrombolysis with a GPIIb/IIIa antagonist and rtPA at reduced dosages is promising but cannot be recommended for general use before prospective randomized clinical trials are completed. PMID- 14752130 TI - Planetary science. Third success reveals a geologic delight. PMID- 14752131 TI - Biomedical research. Senators probe alleged financial conflicts at NIH. PMID- 14752132 TI - California. Budget deepens cuts to UC system. PMID- 14752133 TI - Behavioral ecology. Ecologists roiled by misconduct case. PMID- 14752134 TI - National Science Foundation. Director expected to step down. PMID- 14752135 TI - Infectious diseases. WHO ramps up bird flu vaccine efforts. PMID- 14752136 TI - Remaking NASA. How much space for science? PMID- 14752137 TI - Remaking NASA. Asking for the moon. PMID- 14752138 TI - Remaking NASA. Astronomers ponder a really long-range vision for NASA. PMID- 14752139 TI - Remaking NASA. NASA hopes bigger is better for planned mission to Jupiter. PMID- 14752140 TI - Remaking NASA. NASA's plan for station: from lemon to lemonade. PMID- 14752142 TI - Remaking NASA. From bean counter to visionary: a space odyssey for NASA chief. PMID- 14752141 TI - Remaking NASA. Versatility is the object for new crew vehicle. PMID- 14752143 TI - Conservation policy in coffee landscapes. PMID- 14752145 TI - Comment on "Reactions at interfaces as a source of sulfate formation in sea-salt particles" (I). PMID- 14752144 TI - Cellular distribution of nonionic micelles. PMID- 14752146 TI - Comment on "Reactions at interfaces as a source of sulfate formation in sea-salt particles" (II). PMID- 14752148 TI - Public health. Ethics and the conduct of public health surveillance. PMID- 14752150 TI - Chemistry. Throwing tetrahedral dice. PMID- 14752149 TI - Development. Programming the X chromosome. PMID- 14752151 TI - Astronomy. Nuclear fossils in stardust. PMID- 14752152 TI - Developmental biology. A pattern of precision. PMID- 14752153 TI - The significance of few versus many in the tree of life. PMID- 14752154 TI - Extinct technetium in silicon carbide stardust grains: implications for stellar nucleosynthesis. AB - The isotopic composition of ruthenium (Ru) in individual presolar silicon carbide (SiC) stardust grains bears the signature of s-process nucleosynthesis in asymptotic giant branch stars, plus an anomaly in 99Ru that is explained by the in situ decay of technetium isotope 99Tc in the grains. This finding, coupled with the observation of Tc spectral lines in certain stars, shows that the majority of presolar SiC grains come from low-mass asymptotic giant branch stars, and that the amount of 99Tc produced in such stars is insufficient to have left a detectable 99Ru anomaly in early solar system materials. PMID- 14752155 TI - Elastic torque and the levitation of metal wires by a nematic liquid crystal. AB - Anisotropic particles suspended in a nematic liquid crystal disturb the alignment of the liquid crystal molecules and experience small forces that depend on the particles' orientation. We have measured these forces using magnetic nanowires. The torque on a wire and its orientation-dependent repulsion from a flat surface are quantitatively consistent with theoretical predictions based on the elastic properties of the liquid crystal. These forces can also be used to manipulate submicrometer-scale particles. We show that controlled spatial variations in the liquid crystal's alignment convert the torque on a wire to a translational force that levitates the wire to a specified height. PMID- 14752156 TI - Local gate control of a carbon nanotube double quantum dot. AB - We have measured carbon nanotube quantum dots with multiple electrostatic gates and used the resulting enhanced control to investigate a nanotube double quantum dot. Transport measurements reveal honeycomb charge stability diagrams as a function of two nearly independent gate voltages. The device can be tuned from weak to strong interdot tunnel-coupling regimes, and the transparency of the leads can be controlled independently. We extract values of energy-level spacings, capacitances, and interaction energies for this system. This ability to control electron interactions in the quantum regime in a molecular conductor is important for applications such as quantum computation. PMID- 14752157 TI - An ab initio molecular dynamics study of the aqueous liquid-vapor interface. AB - We present an ab initio molecular dynamics simulation of the aqueous liquid-vapor interface. Having successfully stabilized a region of bulk water in the center of a water slab, we were able to reproduce and further quantify the experimentally observed abundance of surface "acceptor-only"(19%) and "single-donor"(66%) moieties as well as substantial surface relaxation approaching the liquid-vapor interface. Examination of the orientational dynamics points to a faster relaxation in the interfacial region. Furthermore, the average value of the dipole decreases and the average value of the highest occupied molecular orbital for each water molecule increases approaching the liquid-vapor interface. Our results support the idea that the surface contains, on average, far more reactive states than the bulk. PMID- 14752158 TI - Multiferroic BaTiO3-CoFe2O4 Nanostructures. AB - We report on the coupling between ferroelectric and magnetic order parameters in a nanostructured BaTiO3-CoFe2O4 ferroelectromagnet. This facilitates the interconversion of energies stored in electric and magnetic fields and plays an important role in many devices, including transducers, field sensors, etc. Such nanostructures were deposited on single-crystal SrTiO3 (001) substrates by pulsed laser deposition from a single Ba-Ti-Co-Fe-oxide target. The films are epitaxial in-plane as well as out-of-plane with self-assembled hexagonal arrays of CoFe2O4 nanopillars embedded in a BaTiO3 matrix. The CoFe2O4 nanopillars have uniform size and average spacing of 20 to 30 nanometers. Temperature-dependent magnetic measurements illustrate the coupling between the two order parameters, which is manifested as a change in magnetization at the ferroelectric Curie temperature. Thermodynamic analyses show that the magnetoelectric coupling in such a nanostructure can be understood on the basis of the strong elastic interactions between the two phases. PMID- 14752159 TI - Crosstalk between the EGFR and LIN-12/Notch pathways in C. elegans vulval development. AB - The Caenorhabditis elegans vulva is an important paradigm for cell-cell interactions in animal development. The fates of six vulval precursor cells are patterned through the action of the epidermal growth factor receptor-mitogen activated protein kinase (EGFR-MAPK) inductive signaling pathway, which specifies the 1 degrees fate, and the LIN-12/Notch lateral signaling pathway, which specifies the 2 degrees fate. Here, we provide evidence that the inductive signal is spatially graded and initially activates the EGFR-MAPK pathway in the prospective 2 degrees cells. Subsequently, this effect is counteracted by the expression of multiple new negative regulators of the EGFR-MAPK pathway, under direct transcriptional control of the LIN-12-mediated lateral signal. PMID- 14752160 TI - Reactivation of the paternal X chromosome in early mouse embryos. AB - It is generally accepted that paternally imprinted X inactivation occurs exclusively in extraembryonic lineages of mouse embryos, whereas cells of the embryo proper, derived from the inner cell mass (ICM), undergo only random X inactivation. Here we show that imprinted X inactivation, in fact, occurs in all cells of early embryos and that the paternal X is then selectively reactivated in cells allocated to the ICM. This contrasts with more differentiated cell types where X inactivation is highly stable and generally irreversible. Our observations illustrate that an important component of genome plasticity in early development is the capacity to reverse heritable gene silencing decisions. PMID- 14752161 TI - Heterochromatic silencing and HP1 localization in Drosophila are dependent on the RNAi machinery. AB - Genes normally resident in euchromatic domains are silenced when packaged into heterochromatin, as exemplified in Drosophila melanogaster by position effect variegation (PEV). Loss-of-function mutations resulting in suppression of PEV have identified critical components of heterochromatin, including proteins HP1, HP2, and histone H3 lysine 9 methyltransferase. Here, we demonstrate that this silencing is dependent on the RNA interference machinery, using tandem mini-white arrays and white transgenes in heterochromatin to show loss of silencing as a result of mutations in piwi, aubergine, or spindle-E (homeless), which encode RNAi components. These mutations result in reduction of H3 Lys9 methylation and delocalization of HP1 and HP2, most dramatically in spindle-E mutants. PMID- 14752162 TI - Lacticin 481: in vitro reconstitution of lantibiotic synthetase activity. AB - The lantibiotic lacticin 481 is synthesized on ribosomes as a prepeptide (LctA) and posttranslationally modified to its mature form. These modifications include dehydration of serines and threonines, followed by intramolecular addition of cysteines to the unsaturated amino acids, which generates cyclic thioethers. This process breaks eight chemical bonds and forms six newbonds and is catalyzed by one enzyme, LctM. We have characterized the in vitro activity of LctM, which completely processed a series of LctA mutants, displaying a permissive substrate specificity that holds promise for antibiotic engineering. PMID- 14752163 TI - PEST domain-enriched tyrosine phosphatase (PEP) regulation of effector/memory T cells. AB - Protein tyrosine kinases and phosphatases cooperate to regulate normal immune cell function. We examined the role of PEST domain-enriched tyrosine phosphatase (PEP) in regulating T cell antigen-receptor function during thymocyte development and peripheral T cell differentiation. Although normal naive T cell functions were retained in pep-deficient mice, effector/memory T cells demonstrated enhanced activation of Lck. In turn, this resulted in increased expansion and function of the effector/memory T cell pool, which was also associated with spontaneous development of germinal centers and elevated serum antibody levels. These results revealed a central role for PEP in negatively regulating specific aspects of T cell development and function. PMID- 14752164 TI - A predator unmasked: life cycle of Bdellovibrio bacteriovorus from a genomic perspective. AB - Predatory bacteria remain molecularly enigmatic, despite their presence in many microbial communities. Here we report the complete genome of Bdellovibrio bacteriovorus HD100, a predatory Gram-negative bacterium that invades and consumes other Gram-negative bacteria. Its surprisingly large genome shows no evidence of recent gene transfer from its prey. A plethora of paralogous gene families coding for enzymes, such as hydrolases and transporters, are used throughout the life cycle of B. bacteriovorus for prey entry, prey killing, and the uptake of complex molecules. PMID- 14752165 TI - Molecular evolution of the SARS coronavirus during the course of the SARS epidemic in China. AB - Sixty-one SARS coronavirus genomic sequences derived from the early, middle, and late phases of the severe acute respiratory syndrome (SARS) epidemic were analyzed together with two viral sequences from palm civets. Genotypes characteristic of each phase were discovered, and the earliest genotypes were similar to the animal SARS-like coronaviruses. Major deletions were observed in the Orf8 region of the genome, both at the start and the end of the epidemic. The neutral mutation rate of the viral genome was constant but the amino acid substitution rate of the coding sequences slowed during the course of the epidemic. The spike protein showed the strongest initial responses to positive selection pressures, followed by subsequent purifying selection and eventual stabilization. PMID- 14752166 TI - A genome-wide screen identifies genes required for centromeric cohesion. AB - During meiosis, two chromosome segregation phases follow a single round of DNA replication. We identified factors required to establish this specialized cell cycle by examining meiotic chromosome segregation in a collection of yeast strains lacking all nonessential genes. This analysis revealed Sgo1, Chl4, and Iml3 to be important for retaining centromeric cohesin until the onset of anaphase II. Consistent with this role, Sgo1 localizes to centromeric regions but dissociates at the onset of anaphase II. The screen described here provides a comprehensive analysis of the genes required for the meiotic cell cycle and identifies three factors important for the stepwise loss of sister chromatid cohesion. PMID- 14752167 TI - Cross-linking cellular prion protein triggers neuronal apoptosis in vivo. AB - Neuronal death is a prominent, but poorly understood, pathological hallmark of prion disease. Notably, in the absence of the cellular prion protein (PrPC), the disease-associated isoform, PrPSc, appears not to be intrinsically neurotoxic, suggesting that PrPC itself may participate directly in the prion neurodegenerative cascade. Here, cross-linking PrPC in vivo with specific monoclonal antibodies was found to trigger rapid and extensive apoptosis in hippocampal and cerebellar neurons. These findings suggest that PrPC functions in the control of neuronal survival and provides a model to explore whether cross linking of PrPC by oligomeric PrPSc can promote neuronal loss during prion infection. PMID- 14752169 TI - Osteoarthritis--the impact of a serious disease. AB - Osteoarthritis (OA) is common in the elderly, but also affects younger people. The disease symptoms are debilitating and, as well as causing physical impairment, can affect the psychosocial wellbeing of the patient. Furthermore, the impact of this disease is substantially increased by the common occurrence of comorbid conditions, such as hypertension and renal impairment. Non-steroidal anti-inflammatory drugs are commonly used to treat the symptoms of OA, but their related gastrointestinal side-effects increase the impact of this disease. Gastrointestinal tolerability should therefore be considered in the design of new therapies that reduce the symptoms and activity of OA. Furthermore, because this disease is associated with comorbid conditions, patient safety must also be considered when designing new therapies. PMID- 14752170 TI - Osteoarthritis therapy--are there still unmet needs? AB - Non-steroidal anti-inflammatory drugs (NSAIDs) and selective cyclooxygenase (COX) 2 inhibitors are commonly used to control pain and inflammation in osteoarthritis. However, these agents have been associated with gastrointestinal, renal and cardiovascular adverse effects. Together, these complications indicate a clear unmet need in the safety of current treatment options for the management of osteoarthritis. NSAIDs are known to have adverse gastrointestinal effects, and more recently it has been suggested that some selective COX-2 inhibitors are also associated with serious gastrointestinal complications. Selective COX-2 inhibitors have a similar capacity to NSAIDs to delay ulcer healing, and may not significantly decrease the incidences of perforation, ulceration and bleeding (the most clinically relevant gastrointestinal endpoints) compared with NSAIDs. These effects may be due to overlapping roles of COX-1 and COX-2 in physiological and pathophysiological processes. Furthermore, as COX-2 is integrally involved in renal homeostasis, selective COX-2 inhibitors are associated with negative effects on kidney function similar to those seen with NSAIDs. Electrolyte disturbances, oedema and hypertension have been correlated with the use of both drug classes. Additionally, selective COX-2 inhibitors have the potential to increase cardiovascular events, although further research is required to clearly determine such a risk. With the current unmet needs in the treatment of osteoarthritis, the opportunity exists for the development of new therapies. Novel agents include the COX-inhibiting nitric oxide donors and the lipoxygenase (LOX)/COX inhibitor licofelone. Initial results suggest that these therapies may have tolerability advantages over the NSAIDs and selective COX-2 inhibitors. PMID- 14752171 TI - Safety of anti-inflammatory treatment--new ways of thinking. AB - The development of osteoarthritis may be accompanied by increased production of leukotrienes (LTs) and prostaglandins (PGs) from arachidonic acid. These products contribute to joint damage, pain and inflammation. Cyclooxygenase (COX)-1 and COX 2 are responsible for the production of PGs. Inhibition of these enzymes by non steroidal anti-inflammatory drugs and selective COX-2 inhibitors reduces the levels of PGs, resulting in a reduction in pain and inflammation. However, this inhibition can cause alternative processing of arachidonic acid via the 5 lipoxygenase (5-LOX) pathway, resulting in increased production of proinflammatory and gastrotoxic LTs. Licofelone is a competitive inhibitor of 5 LOX, COX-1 and COX-2 that is currently being developed for the treatment of osteoarthritis. Licofelone decreases the production of both LTs and PGs, and thereby reduces inflammation and pain with low gastrotoxicity. Unlike selective COX-2 inhibitors, coadministration of licofelone and aspirin does not appear to be associated with an increase in gastrointestinal adverse events, at least under experimental conditions. Furthermore, there is evidence from animal models to suggest that licofelone may stop disease progression. PMID- 14752172 TI - Licofelone--clinical update on a novel LOX/COX inhibitor for the treatment of osteoarthritis. AB - Licofelone, a competitive inhibitor of 5-lipoxygenase, cyclooxygenase (COX)-1 and COX-2, is currently in clinical development for the treatment of osteoarthritis (OA). Licofelone decreases the production of proinflammatory leukotrienes and prostaglandins-which are involved in the pathophysiology of OA and in gastrointestinal (GI) damage induced by NSAIDs-and has the potential to combine good analgesic and anti-inflammatory effects with excellent GI tolerability. Initial endoscopy data in healthy volunteers have demonstrated that licofelone is well tolerated and has a GI safety profile similar to placebo and significantly better than naproxen. These tolerability results were confirmed in patients with OA in two separate randomized studies. Furthermore, a long-term study (52 weeks) has shown that licofelone is at least as effective as naproxen in the treatment of OA. Licofelone also appears to be as effective as the selective COX-2 inhibitor celecoxib in the treatment of the signs and symptoms of OA. Licofelone has a GI safety profile similar to that of celecoxib, but may offer the advantage of fewer incidences or worsening of peripheral oedema. Preliminary data have also shown that licofelone coadministration with low-dose aspirin does not lead to increased GI toxicity. The emerging clinical data for licofelone indicate that it is an effective and well-tolerated therapy that could offer safety advantages over current treatment options, and that it could be suitable for the long-term treatment of a broad spectrum of patients with OA. PMID- 14752173 TI - How can fetal lung volume be monitored? PMID- 14752174 TI - Technology assessment for radiologists. AB - Health technology assessment is the systematic and quantitative evaluation of the safety, efficacy, and cost of health care interventions. This article outlines aspects of technology assessment of diagnostic imaging. First, it presents a conceptual framework of a hierarchy of levels of efficacy that should guide thinking about imaging test evaluation. In particular, the framework shows how the question answered by most evaluations of imaging tests, "How well does this test distinguish disease from the nondiseased state?" relates to the fundamental questions for all health technology assessment, "How much does this intervention improve the health of people?" and "What is the cost of that improvement?" Second, it describes decision analysis and cost-effectiveness analysis, which are quantitative modeling techniques usually used to answer the two core questions for imaging. Third, it outlines design and operational considerations that are vital if researchers who are conducting an experimental study are to make a quality contribution to technology assessment, either directly through their findings or as an input into decision analyses. Finally, it includes a separate discussion of screening--that is, the application of diagnostic tests to nonsymptomatic populations--because the requirements for good screening tests are different from those for diagnostic tests of symptomatic patients and because the appropriate evaluation methods also differ. PMID- 14752175 TI - Nanotechnology and medicine. PMID- 14752176 TI - Quality and consistency in CT colonography and research reporting. PMID- 14752178 TI - CT angiography for diagnosis of pulmonary embolism: state of the art. AB - In daily clinical routine, computed tomography (CT) has practically become the first-line modality for imaging of pulmonary circulation in patients suspected of having pulmonary embolism (PE). However, limitations regarding accurate diagnosis of small peripheral emboli have so far prevented unanimous acceptance of CT as the reference standard for imaging of PE. The development of multi-detector row CT has led to improved visualization of peripheral pulmonary arteries and detection of small emboli. The finding of a small isolated clot at pulmonary CT angiography, however, may be increasingly difficult to correlate with results of other imaging modalities, and the clinical importance of such findings is uncertain. Therefore, the most realistic scenario to measure efficacy of pulmonary CT angiography when PE is suspected may be assessment of patient outcome. Meanwhile, the high negative predictive value of a normal pulmonary CT angiographic study and its association with beneficial patient outcome has been demonstrated. While the introduction of multi-detector row technology has improved CT diagnosis of PE, it has also challenged its users to develop strategies for optimized contrast material delivery, reduction of radiation dose, and management of large-volume data sets created at those examinations. PMID- 14752179 TI - Late-stage adult respiratory distress syndrome caused by severe acute respiratory syndrome: abnormal findings at thin-section CT. AB - PURPOSE: To evaluate thin-section computed tomographic (CT) abnormalities in patients in the intensive care unit during the late stage of adult respiratory distress syndrome (ARDS) caused by severe acute respiratory syndrome (SARS). MATERIALS AND METHODS: Eight patients in the late stage of ARDS (ie, more than 2 weeks after onset) were imaged with thin-section CT. Images were evaluated for ground-glass opacification, consolidation, interstitial thickening, evidence of fibrosis, and any other abnormalities. Patient records were reviewed, and relevant respiratory and ventilatory parameters, total steroid dose, and outcome were recorded. RESULTS: All patients received high-dose pulse methylprednisolone (minimum, 2.5 g total), and all patients who received ventilation received low pressure, low-volume ventilation. Five patients received prolonged mechanical ventilation (for more than 14 days), one received ventilation for 72 hours, and two patients did not receive ventilation. Three patients died, four were discharged from the hospital, and one continued to require ventilation. Ground glass opacification and interstitial thickening were present at CT in all eight patients. Consolidation was present in six patients. Three patients had evidence of fibrosis. Patients who received long-term ventilation, those who received short-term ventilation, and those who did not receive ventilation had similar pulmonary changes at CT. Pulmonary cysts, most of which were small (<1 cm), were present in five patients. Cysts were present in one patient who received only short-term low-pressure and low-volume ventilation and in one patient who received no mechanical ventilation. CONCLUSION: The CT features of late-stage ARDS caused by SARS are similar to those seen in late-stage ARDS of other causes, with no apparent differences between patients who do and patients who do not receive prolonged mechanical ventilation. The presence of cysts in one patient who received short-term and one patient who received no mechanical ventilation suggests that severe SARS-induced ARDS may independently result in cyst formation. PMID- 14752180 TI - Pulmonary nodules at chest CT: effect of computer-aided diagnosis on radiologists' detection performance. AB - PURPOSE: To evaluate the effect of computer-aided diagnosis (CAD) on radiologists' detection of pulmonary nodules. MATERIALS AND METHODS: Fifty chest computed tomographic (CT) examination cases were used. The mean nodule size was 0.81 cm +/- 0.60 (SD) (range, 0.3-2.9 cm). Alternative free-response receiver operating characteristic (ROC) analysis with a continuous rating scale was used to compare the observers' performance in detecting nodules with and without use of CAD. Five board-certified radiologists and five radiology residents participated in an observer performance study. First they were asked to rate the probability of nodule presence without using CAD; then they were asked to rate the probability of nodule presence by using CAD. RESULTS: For all radiologists, the mean areas under the best-fit alternative free-response ROC curves (Az) without and with CAD were 0.64 +/- 0.08 and 0.67 +/- 0.09, respectively, indicating a significant difference (P <.01). For the five board-certified radiologists, the mean Az values without and with CAD were 0.63 +/- 0.08 and 0.66 +/- 0.09, respectively, indicating a significant difference (P <.01). For the five resident radiologists, the mean Az values without and with CAD were 0.66 +/- 0.04 and 0.68 +/- 0.04, respectively, indicating a significant difference (P =.02). At observer performance analyses, there were no significant differences in Az values obtained either without (P =.61) or with (P =.88) CAD between the board certified radiologists and the residents. For all radiologists, in the detection of pulmonary nodules 1.0 cm in diameter or smaller, the mean Az values without and with CAD were 0.60 +/- 0.11 and 0.64 +/- 0.11, respectively, indicating a significant difference (P <.01). CONCLUSION: Use of the CAD system improved the board-certified radiologists' and residents' detection of pulmonary nodules at chest CT. PMID- 14752181 TI - Percutaneous MR imaging-guided radiofrequency interstitial thermal ablation of tongue base in porcine models: implications for obstructive sleep apnea syndrome. AB - PURPOSE: To test the feasibility and safety of a percutaneous magnetic resonance (MR) imaging-guided technique for radiofrequency (RF) interstitial thermal ablation of the tongue base and to correlate MR appearance of induced thermal lesions with histopathologic findings in pigs in acute and chronic porcine models. MATERIALS AND METHODS: A 1-cm-tip RF electrode was inserted percutaneously into the tongue in 10 pigs with 0.2-T real-time MR guidance. The RF electrode was advanced up the midline between lingual arteries and stopped short of tongue mucosa. RF interstitial thermal ablation was performed at 90 degrees C +/- 2 and lasted 10 minutes. Postablation images were obtained with a 1.5-T MR imager. Five pigs were sacrificed immediately (acute model), while five were followed up for 1 month (chronic model) before they were sacrificed. MR compatible fiducial coils were inserted into tongues with MR imaging guidance prior to RF ablation in the chronic group. Tongues were harvested for histopathologic analysis. Mean thermal lesion volume was compared with the Student t test on images obtained immediately, 2 weeks, and 1 month after RF ablation. Interclass correlation coefficients of lesion diameters at gross pathologic analysis and corresponding diameters with each pulse sequence were calculated. RESULTS: Successful MR imaging-guided electrode positioning was achieved in all procedures without intra- or postprocedure complications because there was high vascular conspicuity and tissue contrast. Thermal lesions appeared hypointense with hyperintense surrounding rims with all sequences in both groups. At pathologic analysis, acute lesions appeared as pale necrotic areas surrounded by hyperemic rims, while chronic lesions demonstrated progressive circumferential fibrosis and significant volume shrinkage (P <.01). Thermal lesion diameters measured at gross pathologic analysis best agreed with corresponding diameters measured on short inversion time inversion-recovery images (interclass correlation coefficient = 0.85). CONCLUSION: The results of this investigation demonstrate MR imaging-guided RF interstitial thermal ablation of the tongue base is feasible and safe and illustrate imaging and pathologic phenomena associated with creation and evolution of the induced thermal lesions. PMID- 14752182 TI - Bone formation in transplants of human bone marrow stromal cells and hydroxyapatite-tricalcium phosphate: prediction with quantitative CT in mice. AB - PURPOSE: To determine whether quantitative computed tomography (CT) can be used to estimate the extent of new bone formation in hydroxyapatite-tricalcium phosphate (HA-TCP)-based transplants. MATERIALS AND METHODS: Bone-forming transplants were generated by attaching cultured human bone marrow stromal cells to aliquots of HA-TCP particles and were placed in subcutaneous pockets in immunocompromised mice. After 8 weeks, the transplants were individually imaged; each scan included a phantom. Overall bone mineral density (BMD) of each transplant was obtained. Hematoxylin-eosin-stained sections of the same transplants were then examined histologically, which is the reference standard for assessing bone formation. The extent of bone in each transplant was scored on a semiquantitative scale ranging from 0 to 4 by three independent blinded observers; the bone score for each transplant was calculated by averaging the three observer scores. BMD was compared with the histologically determined bone score for each transplant. Statistical evaluations included (a) calculation of empiric receiver operating characteristic curves to determine optimum BMD thresholds and (b) determination of the relationship between BMD and bone score, including derivation of Pearson correlation coefficients. RESULTS: One hundred twenty transplants were evaluated. Average BMD of 600 mg/cm3 K2HPO4 or more was noted in transplants with appreciable bone formation (bone score > or = 3), while average BMD of less than 600 mg/cm3 K2HPO4 was seen in transplants with poor bone formation (bone score < 3) (P <.001). Among transplants with appreciable bone formation, the BMD was proportional to the extent of mineralized matrix present in the new bone. CONCLUSION: Use of quantitative CT offers a practical approach for the noninvasive determination of new bone formation in mineralizing bone marrow stromal cells and HA-TCP transplants. PMID- 14752183 TI - Coronary artery calcium quantification at multi-detector row helical CT versus electron-beam CT. AB - PURPOSE: To compare coronary artery calcium scores from a multi-detector row helical computed tomographic (CT) scanner with those from an electron-beam CT scanner, with emphasis on subjects with calcium scores less than 400. MATERIALS AND METHODS: Seventy-eight asymptomatic subjects (37 women, 41 men; age range, 39 78 years; mean age, 54.2 years) underwent multi-detector row CT and electron-beam CT. Volume and Agatston scores were calculated with a workstation. Statistical analyses included assessment of association between calcium scores from two scanners, calculation of percent absolute difference to assess score variability between scanners, equivalence analysis, construction of Bland-Altman plots to assess agreement between scores, and assessment of changes in score grouping and risk criteria based on score differences between scanners. RESULTS: Electron-beam CT calcium scores were higher than multi-detector row CT scores. Linear association between calcium scores obtained from paired scans was significant (r = 0.96-0.99, P <.001). Mean percent absolute differences were 67.9% and 65.0% for volume and Agatston scores, respectively (48.6% and 46.3% for corresponding natural log-transformed scores). In subjects with a score of 11 or greater, mean percent absolute differences between electron-beam CT and multi-detector row CT scores ranged from 15% to 30% (<10% for natural log-transformed calcium scores). With a 20% equivalence limit, calcium scores from the two scanners were statistically equivalent (P <.05). Score grouping would have been subject to change in 12 (11 increased and one decreased; six with scores of 11 or greater), and possible risk management decisions would have been subject to change in eight (16%) of 51 subjects who underwent electron-beam CT versus multi-detector row CT scanning. CONCLUSION: Multi-detector row CT appears to be comparable to electron beam CT for coronary calcification screening, except in subjects with a calcium score less than 11. PMID- 14752185 TI - Case 67: Persistent stapedial artery. PMID- 14752186 TI - Pheochromocytomas: detection with 11C hydroxyephedrine PET. AB - PURPOSE: To evaluate the accuracy of carbon 11 (11C) hydroxyephedrine (HED) positron emission tomography (PET) in the detection of pheochromocytomas. MATERIALS AND METHODS: Nineteen patients (12 women, seven men; mean age, 53 years) suspected of having pheochromocytoma were evaluated. Patients had enlarged adrenal glands at computed tomography and either increased urinary catecholamine levels (n = 18) or normal biochemistry (n = 1). Dynamic PET examination in the adrenal region was performed after injection of 800 MBq 11C HED. PET data were analyzed visually and semiquantitatively. Time-activity curves were generated for different organs. PET results were validated with histologic evaluation (n = 16) or clinical follow-up (n = 3). The diagnostic value of HED PET was evaluated by calculating the sensitivity, specificity, positive predictive value, negative predictive value, and accuracy. RESULTS: In 12 patients, 13 pheochromocytomas were verified at surgery and histologic evaluation. All but one of the pheochromocytomas were detected with HED PET, which demonstrated elevated uptake. The rest of the patients (n = 7) did not have pheochromocytomas. In these patients, HED PET did not show any abnormal uptake in the suspicious tumors (confirmed at surgery in four patients and at clinical follow-up in three). Mean standardized uptake value of the tumors was 21.4 (range, 11.1-40.9). The time activity curves for pheochromocytomas showed early uptake after injection, and the activity increased with the time of examination. Sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of HED PET in the detection of pheochromocytomas were 92% (12 of 13), 100% (seven of seven), 100% (12 of 12), 87.5% (seven of eight), and 95% (19 of 20), respectively. CONCLUSION: HED PET is useful in the detection of pheochromocytomas, providing a high level of accuracy. PMID- 14752187 TI - Precaval right renal arteries: prevalence and morphologic associations at spiral CT. AB - PURPOSE: To determine the prevalence and morphologic associations of precaval right renal arteries at spiral computed tomography (CT). MATERIALS AND METHODS: The authors retrospectively reviewed 186 arterial phase contrast material enhanced spiral CT scans of the abdomen (5.0-mm section thickness in 97 scans, 2.5 mm in 89 scans) obtained during a 2-year period to identify patients with precaval right renal arteries. During routine interpretation of CT scans at daily readout, the authors prospectively identified 39 additional patients with precaval right renal arteries. All cases were evaluated for anatomic variants and associated clinical findings. Fisher exact analysis and chi2 analysis were performed to compare the frequency of anatomic variants between patients with and those without precaval renal arteries. RESULTS: Nine of 186 patients had precaval right renal arteries, for a prevalence of 5%. In the 48 patients with precaval renal arteries, 52 precaval arteries were found, of which 48 were accessory and four were dominant. Fourteen patients had right pelviectasis to the level of the precaval artery, and three of these had a clinical diagnosis of right ureteropelvic junction obstruction. Eighteen (35%) of the 52 precaval renal arteries arose from the anterior aspect of the aorta (within 30 degrees of the midline). The lower pole of the right kidney was rotated anteriorly in two (22%) of nine and 13 (33%) of 39 patients with precaval renal arteries in the retrospective and prospective groups, respectively, compared with four (2%) of 177 patients without precaval arteries (P <.05 and P <.001, respectively). CONCLUSION: On the basis of these results, precaval right renal arteries appear to be more common than previously reported. Anterior rotation of the lower pole of the right kidney should prompt a search for precaval renal arteries. PMID- 14752188 TI - Preoperative staging of gastric adenocarcinoma: comparison of helical CT and endoscopic US. AB - PURPOSE: To compare the performance of helical computed tomography (CT) and endoscopic ultrasonography (US) in the preoperative staging of gastric cancer. MATERIALS AND METHODS: Fifty-one consecutive patients with a primary malignant gastric tumor (stage T2-T4) were preoperatively evaluated with both helical CT and endoscopic US within 3 days. Each tumor was staged according to the TNM classification system with both modalities. All patients subsequently underwent surgery. Results of CT and endoscopic US were compared with histologic staging of tumor invasion depth and regional lymph node metastasis. For comparison of CT and endoscopic US data, the marginal homogeneity test was used, and a P value of less than.05 was determined to indicate statistical significance. RESULTS: In comparison with histologic results, CT achieved correct T staging in 39 patients (76%) and correct N staging in 35 patients (70%). The corresponding results for endoscopic US achieved correct T staging in 44 patients (86%) and correct N staging in 45 patients (90%). There was no significant difference between T staging (P =.55) and N staging (P >.99). Because of challenging detection of wall layers, correct T staging was difficult for CT and endoscopic US in the differentiation of T2 and T3 lesions. CONCLUSION: Compared with endoscopic US, helical CT focused on the stomach provides valuable results regarding T and N staging in patients with gastric cancer. PMID- 14752189 TI - Littoral cell angioma of the spleen: CT features with clinicopathologic comparison. AB - PURPOSE: To evaluate the clinical, pathologic, and computed tomographic (CT) features of littoral cell angioma of the spleen in eight patients. MATERIALS AND METHODS: Two abdominal radiologists retrospectively reviewed the contrast material-enhanced CT images obtained in six, the contrast-enhanced and nonenhanced CT images obtained in two, and the photographs of gross pathologic specimens resected from seven patients. They also retrospectively reviewed clinical data (ie, demographic data, presenting signs and symptoms, physical findings, and medical histories). Histopathologic specimens from the eight patients were reviewed by a hematopathologist. The CT images were reviewed for the presence of splenomegaly. The number, size, and enhancement characteristics of the splenic masses at CT were compared with the histopathologic and gross pathologic specimen findings. RESULTS: All patients had laboratory evidence of hypersplenism. Seven patients (88%) had splenomegaly and innumerable splenic masses ranging from 0.2 to 6.0 cm in diameter at CT. The single patient with a normal spleen size had four splenic masses. The splenic masses were hypoattenuating relative to the normal spleen at CT in all patients and correlated with blood-filled nodules at gross pathologic examination and with blood-filled vascular channels of littoral cell angioma at histopathologic examination. The early and late portal venous phase CT images that were available in one case demonstrated progressive homogeneous contrast enhancement of the masses such that they were indistinguishable from the normal splenic parenchyma. CONCLUSION: Littoral cell angioma is a primary splenic neoplasm that most commonly manifests at CT as multiple hypoattenuating masses in an enlarged spleen. Histopathologically, these masses represent blood-filled vascular channels. PMID- 14752190 TI - The rugger jersey spine sign. PMID- 14752191 TI - Intervertebral disk calcification of the spine in an elderly population: radiographic prevalence, location, and distribution and correlation with spinal degeneration. AB - PURPOSE: To determine the prevalence, distribution, and location of intervertebral disk calcification (IDC) in the thoracic and lumbar spine and the association of IDC with radiographically evident spinal degenerative changes in cadavers. MATERIALS AND METHODS: Anterior vertebral columns comprising T1 through L5 were removed from 223 cadavers (183 men, 40 women; mean age at death, 67 years; range, 37-94 years). Approximately 5-mm-thick parasagittal sections were investigated with high-contrast radiography. The presence of IDC, osteophytes, vertebral endplate abnormalities, and vacuum phenomena was recorded, and the height of disk space was measured at 3,568 intervertebral levels. Logistic regression analysis was performed. RESULTS: IDC was identified in 178 (80%) of the 223 cadavers. Of 3,568 disks, 459 (13%) had IDC, and 289 cases (63%) were located in the annulus fibrosus. IDC was most common in the lower thoracic spine, occurring in 275 (60%) of 459 disks. IDC occurred in 159 (87%) of 183 men and 19 (48%) of 40 women. Logistic regression analysis was adjusted for age, and results showed that the frequency of IDC was significantly higher in men in upper, middle, and lower segments of the thoracic spine (P <.05) but not in the lumbar spine (P =.09). IDC correlated with increasing age (P <.001) and disk space loss (P <.001) at all spinal levels. There was no association of IDC with vacuum phenomena or vertebral endplate abnormalities at any spinal level. CONCLUSION: IDC is common in elderly persons, especially in the annulus fibrosus and lower thoracic spine. The prevalence of IDC increases with age and extent of disk space loss. PMID- 14752192 TI - Pattern of white matter abnormalities at MR imaging: use of polymerase chain reaction testing of Guthrie cards to link pattern with congenital cytomegalovirus infection. AB - PURPOSE: To define a magnetic resonance (MR) imaging pattern suggestive of congenital cytomegalovirus (CMV) infection by using polymerase chain reaction (PCR) testing to detect CMV DNA in neonatal blood on Guthrie cards for validation. MATERIALS AND METHODS: On the basis of findings in eight patients with documented congenital CMV infection, the authors developed MR imaging inclusion criteria, including multifocal lesions predominantly located in the deep parietal white matter. If gyral abnormalities were present, white matter lesions were either multifocal or diffuse. The criteria were applied to 152 patients with static leukoencephalopathy of unknown etiology. Guthrie cards for 22 of the 43 patients fulfilling the MR imaging criteria, 20 patients not fulfilling them, and 300 control subjects were analyzed. Fisher exact testing was used to evaluate the association between MR imaging characteristics and CMV status, and backward elimination linear discriminant analysis was used to identify MR imaging characteristics predictive of CMV infection in addition to the initial criteria. RESULTS: PCR test results were positive in 12 of 22 patients suspected of having congenital CMV infection, in no patient not suspected of having infection (P <.001), and in two of 300 control subjects (negative predictive value [NPV] of MR imaging criteria, 100% [95% CI: 83%, 100%]; positive predictive value [PPV], 55% [95% CI: 32%, 76%]). The most important additional MR imaging finding predicting a positive PCR result was abnormality of the anterior part of the temporal lobe, including abnormal white matter, cysts, and enlargement of inferior horns. Including this finding in the MR imaging criteria enhanced the PPV (89%; 95% CI: 52%, 99%) at the expense of the NPV (88%; 95% CI: 72%, 97%). CONCLUSION: In patients with static encephalopathy, an MR imaging pattern of multifocal lesions predominantly involving deep parietal white matter, with or without gyral abnormalities, is predictive of congenital CMV infection. When gyral abnormalities are present, leukoencephalopathy may also be diffuse. The presence of abnormalities in the anterior part of the temporal lobe increases the likelihood that CMV infection is present. PMID- 14752193 TI - Screening-detected breast cancer in a man with BRCA2 mutation: case report. AB - Current indications for breast cancer screening in the male population are lacking, although family history of breast cancer may be such an indication. The authors describe a man with a history of clinically diagnosed right breast cancer who subsequently tested positive for the breast cancer susceptibility gene BRCA2 and received a diagnosis of mammographically detected left breast cancer at screening. The authors discuss the clinical implications of this approach to detecting male breast cancer. PMID- 14752194 TI - Endovascular brachytherapy: effect on acute inflammatory response after percutaneous femoropopliteal arterial interventions. AB - PURPOSE: To investigate whether endovascular brachytherapy diminishes vascular inflammation in response to femoropopliteal percutaneous transluminal angioplasty (PTA) or stent implantation in two double-blind randomized-controlled trials. MATERIALS AND METHODS: Forty-seven consecutive patients from two double-blind randomized-controlled trials were studied. Patients either underwent femoropopliteal PTA with endovascular gamma irradiation (n = 8) or placebo irradiation (n = 7) or underwent PTA and stent implantation with brachytherapy (n = 15) or placebo irradiation (n = 17). High-sensitivity C-reactive protein (CRP), serum amyloid A (SAA), and fibrinogen levels were measured at baseline and 8, 24, and 48 hours after the intervention. The change of acute phase parameters from baseline to 48 hours after intervention indicated the extent of the inflammatory response and was compared between patients undergoing brachytherapy and those undergoing placebo irradiation. Fisher exact test was used for comparison of categorical data, and nonparametric statistical methods were applied for analysis of continuous data (Mann-Whitney U tests for unpaired data and Friedman analysis for repetitive measurements). RESULTS: Median patient age was 70 years (interquartile range, 56-74 years); 33 (70%) patients were men and 14 (30%) were women. Clinical characteristics and baseline values of acute phase parameters were similar between groups. A statistically significant increase in CRP, SAA, and fibrinogen values was observed after PTA and stent implantation, both in the patients who underwent brachytherapy and in those who underwent placebo irradiation. Compared with placebo irradiation, however, brachytherapy did not significantly reduce any acute phase parameter from baseline to 8, 24, or 48 hours after the intervention (P >.05 for all comparisons). CONCLUSION: Endovascular brachytherapy did not diminish early vascular inflammation in response to PTA or stent implantation and even induced a trend toward an increased inflammatory response. PMID- 14752195 TI - Dark flow artifacts with steady-state free precession cine MR technique: causes and implications for cardiac MR imaging. AB - Steady-state free precession cine images from cardiac magnetic resonance imaging studies of 24 patients were reviewed retrospectively to identify dark flow artifacts. The cause and features of the artifacts were studied in flow phantom experiments. Dark flow artifacts were recognized in eight of the 24 cases and were characterized by low or inhomogeneous signal intensity in blood pools with little change in adjacent tissues. The artifacts could be mimicked in flow phantom experiments by deliberately deshimming the gradients and appeared periodically during imaging with off-centered frequencies. These artifacts appeared to be caused by spins moving within an inhomogeneous magnetic field. PMID- 14752196 TI - Long-term retention of mammographic computer-assisted diagnosis information is neither necessary nor desirable. PMID- 14752197 TI - Diagnosis please comments. PMID- 14752198 TI - Automated protein crystal structure determination using ELVES. AB - Efficient determination of protein crystal structures requires automated x-ray data analysis. Here, we describe the expert system ELVES and its use to determine automatically the structure of a 12-kDa protein. Multiwavelength anomalous diffraction analysis of a selenomethionyl derivative was used to image the Asn-16 Ala variant of the GCN4 leucine zipper. In contrast to the parallel, dimeric coiled coil formed by the WT sequence, the mutant unexpectedly formed an antiparallel trimer. This structural switch reveals how avoidance of core cavities at a single site can select the native fold of a protein. All structure calculations, including indexing, data processing, locating heavy atoms, phasing by multiwavelength anomalous diffraction, model building, and refinement, were completed without human intervention. The results demonstrate the feasibility of automated methods for determining high-resolution, x-ray crystal structures of proteins. PMID- 14752199 TI - Structures of the N-terminal modules imply large domain motions during catalysis by methionine synthase. AB - B(12)-dependent methionine synthase (MetH) is a large modular enzyme that utilizes the cobalamin cofactor as a methyl donor or acceptor in three separate reactions. Each methyl transfer occurs at a different substrate-binding domain and requires a different arrangement of modules. In the catalytic cycle, the cobalamin-binding domain carries methylcobalamin to the homocysteine (Hcy) domain to form methionine and returns cob(I)alamin to the folate (Fol) domain for remethylation by methyltetrahydrofolate (CH(3)-H(4)folate). Here, we describe crystal structures of a fragment of MetH from Thermotoga maritima comprising the domains that bind Hcy and CH(3)-H(4)folate. These substrate-binding domains are (beta alpha)(8) barrels packed tightly against one another with their barrel axes perpendicular. The properties of the domain interface suggest that the two barrels remain associated during catalysis. The Hcy and CH(3)-H(4)folate substrates are bound at the C termini of their respective barrels in orientations that position them for reaction with cobalamin, but the two active sites are separated by approximately 50 A. To complete the catalytic cycle, the cobalamin binding domain must travel back and forth between these distant active sites. PMID- 14752200 TI - Notch is required for long-term memory in Drosophila. AB - A role for Notch in the elaboration of existing neural processes is emerging that is distinct from the increasingly well understood function of this gene in binary cell-fate decisions. Several research groups, by using a variety of organisms, have shown that Notch is important in the development of neural ultrastructure. Simultaneously, Presenilin (Psn) was identified both as a key mediator of Notch signaling and as a site of genetic lesions that cause early-onset Alzheimer's disease. Here we demonstrate that Notch loss of function produces memory deficits in Drosophila melanogaster. The effects are specific to long-term memory, which is thought to depend on ultrastructural remodeling. We propose that Notch plays an important role in the neural plasticity underlying consolidated memory. PMID- 14752203 TI - Top-down approaches to the study of natural variation in complex physiological pathways using the white-footed mouse (Peromyscus leucopus) as a model. AB - Variation in complex physiological pathways has important effects on human function and medical treatment. Complex pathways involve cells at multiple locations, which serve different functions regulated by many genes and include complex neuroendocrine pathways that regulate physiological function. One of two competing hypotheses regarding the effects of selection on complex pathways predicts that variability should be common within complex pathways. If this hypothesis is correct, then we should expect wide variation in neuroendocrine function to be typical within natural populations. To test this hypothesis, a complex neuroendocrine pathway that regulates photoperiod-dependent changes in fertility in a natural population of white-footed mice (Peromyscus leucopus) was used to test for natural genetic variability in multiple components of the pathway. After testing only six elements in the photoperiod pathway in P. leucopus, genetic variation in the following four of these elements was evident: the circadian clock, melatonin receptor abundance or affinity, sensitivity of the reproductive axis to steroid negative feedback, and gonadotropin-releasing hormone neuronal activity. If this result can be extended to humans, the prediction would be that significant variation at multiple loci in complex neuroendocrine pathways is common among humans, and that variation would exist even in human populations from a common genetic background. This finding could only be drawn from an "exotic" animal model derived from a natural source population, confirming the continuing importance of nontraditional models alongside the standard laboratory species. PMID- 14752204 TI - Octodon degus: a diurnal, social, and long-lived rodent. AB - Octodon degus is a moderate-sized, precocious, but slowly maturing, hystricomorph rodent from central Chile. We have used this species to study a variety of questions about circadian rhythms in a diurnal mammal that readily adapts to most laboratory settings. In collaboration with others, we have found that a number of fundamental features of circadian function differ in this diurnal rodent compared with nocturnal rodents, specifically rats or hamsters. We have also discovered that many aspects of the circadian system are sexually dimorphic in this species. However, the sexual dimorphisms develop in the presence of pubertal hormones, and the sex differences do not appear until after gonadal puberty is complete. The developmental timing of the sex differences is much later than in the previously studied altricial, rapidly developing rat, mouse, or hamster. This developmental timing of circadian function is reminiscent of that reported for adolescent humans. In addition, we have developed a model that demonstrates how nonphotic stimuli, specifically conspecific odors, can interact with the circadian system to hasten recovery from a phase-shift of the light:dark cycle (jet lag). Interestingly, the production of the odor-based social signal and sensitivity to it are modulated by adult gonadal hormones. Data from degu circadian studies have led us to conclude that treatment of some circadian disorders in humans will likely need to be both age and gender specific. Degus will continue to be valuable research animals for resolving other questions regarding reproduction, diabetes, and cataract development. PMID- 14752205 TI - The musk shrew (Suncus murinus): a model species for studies of nutritional regulation of reproduction. PMID- 14752206 TI - The prairie vole (Microtus ochrogaster): an animal model for behavioral neuroendocrine research on pair bonding. AB - Pair bond formation has been investigated much less than many other social behaviors, perhaps in part because traditional laboratory mice and rats do not exhibit this behavior. However, pair bonding is common among monogamous animals such as the prairie vole (Microtus ochrogaster). In this review, we discuss how the prairie vole has been used as a model system to investigate the neurobiology of pair bonding. Descriptions include neuroanatomical differences between monogamous and non-monogamous voles, as well as how manipulations of vasopressin, oxytocin, dopamine, and corticosterone systems affect pair bond formation. Also summarized are potential interactions among these systems that regulate pair bonding, and the extent of sexual dimorphism in underlying mechanisms. Pair bonding in prairie voles is an excellent model system for studying central processing of social information. Understanding the mechanisms underlying this behavior may provide important insights into human disorders associated with impaired social functioning. PMID- 14752207 TI - Tracing the evolution of brain and behavior using two related species of whiptail lizards: Cnemidophorus uniparens and Cnemidophorus inornatus. AB - Cnemidophorus whiptail lizards offer a unique opportunity to study behavioral and neural evolution because unlike most genera, ancestral and descendant species are still extant, and comparisons between species provide a window into correlated changes in biological organization through speciation. This review focuses on the all-female or parthenogenetic species Cnemidophorus uniparens (descendant species), which evolved through several hybridization events involving the sexually reproducing species Cnemidophorus inornatus (ancestral species). Data compiled over more than 2 decades include behavioral, endocrine, and neural differences between these two related species of whiptail lizards. For example, unlike females of the ancestral species, individuals of the descendant species display male-like mounting behavior (pseudocopulatory behavior) after ovulation. Pseudocopulatory behavior in the parthenogen is triggered by the progesterone surge after ovulation, and the behavioral capacity to respond to progesterone appears to be an ancestral trait that was inherited from C. inornatus males through the hybridization events. Interestingly, the regulation of sex steroid hormone receptor mRNA in brain areas critical for the expression of sociosexual behaviors differs between females of the two species and suggests that evolutionary changes in the regulation of gene expression could be a proximate mechanism that underlies the evolution of a novel social behavior in the parthenogen. Finally, because the sexual species is diploid, whereas the parthenogen is triploid, differences between the species could directly assess the effect of ploidy. The behavioral and neuroendocrinological data are pertinent for considering this possibility. PMID- 14752208 TI - Tall stature, insulin resistance, and disturbed behavior in a girl with the triple X syndrome harboring three SHOX genes: offspring of a father with mosaic Klinefelter syndrome but with two maternal X chromosomes. AB - AIMS: To describe the tall stature and its possible underlying mechanism in a Caucasian girl (age 12 years and 10 months) with 46,XX (28%)/47,XXX (72%) mosaicism and to identify the parental origin of her extra X chromosome. METHODS: The fasting glucose-to-insulin ratio was studied. The karyotypes of the girl and her parents as well as the presence of SHOX copies and the parental origin of her extra X chromosome were assessed. RESULTS: Clinical examination revealed a tall stature and severe acne, and endocrinological/metabolic assessment revealed insulin resistance. Fluorescence in situ hybridization cytogenetic analysis depicted the presence of three SHOX genes in the 47,XXX cell line of the patient. Karyotyping of her parents showed a normal 46,XX karyotype in the mother and 46,XY(93%)/47,XXY(7%) Klinefelter mosaicism in the father. However, DNA analysis unequivocally showed maternal origin of the extra X chromosome of the patient. CONCLUSIONS: This report suggests that SHOX gene triplication may produce a tall stature, even in the presence of preserved ovarian function. X triplication might predispose to insulin resistance and behavioral disorders. PMID- 14752209 TI - Effect of growth hormone replacement therapy on pituitary hormone secretion and hormone replacement therapies in GHD adults. AB - OBJECTIVE: We tested the impact of commencement of GH replacement therapy in GH deficient (GHD) adults on the circulating levels of other anterior pituitary and peripheral hormones and the need for re-evaluation of other hormone replacement therapies, especially the need for dose changes. METHODS: 22 GHD patients were investigated in a double-blind randomized study and 90 GHD patients in an open study at baseline and after 6 and 12 months of GH replacement therapy. RESULTS: In the placebo-controlled trial, the FT(3) levels increased after 6 months in the GH-treated group, and in the open study the FT(3) levels tended to increase. Other hormone concentrations did not change in either part of the study. Four patients required an increase in thyroxine dose, while 2 patients needed dose reduction. One originally euthyroid patient required thyroxine replacement. Two patients with originally conserved pituitary-adrenal function developed ACTH insufficiency. The hydrocortisone dose was increased in 1 and decreased in 1 of the 66 patients with secondary hypocortisolism. None of the females required any adjustment of sex hormone replacement therapy. Two of 37 males needed dose increase of testosterone, while 1 needed dose reduction. CONCLUSION: GH replacement therapy required dose adjustments regarding other hormone replacement therapies in 12.2% (n = 11), while initiation of new hormone replacement was performed in 3.3% (n = 3) of the 90 patients during the 1-year follow-up. Monitoring of pituitary hormone axes is advisable after commencement of GH replacement therapy, since changes of hormone replacement therapy was observed in a small but clinically significant number of patients. PMID- 14752210 TI - Effect of puberty on the relationship between bone markers of turnover and bone mineral density in Turner's syndrome. AB - It has been suggested that the appropriate timing of puberty is necessary for normal bone mineral acquisition which may not be achieved amongst patients with Turner's syndrome (TS). The aim of this study was to assess bone mineral density (BMD) and bone turnover in 34 patients with TS (age range 2.2-39.0 years). The areal BMD (aBMD) was determined by dual-energy X-ray absorptiometry, and the volumetric BMD was calculated. Blood and second voided urine samples were taken the morning after an overnight fast for evaluation of the biochemical markers of bone turnover: bone-specific alkaline phosphatase (BAP) and N-telopeptides of type I collagen (NTX), respectively. Both were determined by enzyme-linked immunosorbent assay. The patients were divided into three groups: group 1 (n = 13; prepubertal; age range 2.2-19.0 years), group 2 (n = 10; teenagers; age range 12.4-19.0 years), and group 3 (n = 11; adults; chronological age >20 years). They were also grouped by breast development according to Tanner stage into B1 (n = 12), B2-3 (n = 9), and B4-5 (n = 13). The aBMD was significantly lower in group 1 and was higher at Tanner stages 4 and 5 as compared with patients at Tanner stage 1. The bone turnover markers were significantly higher in group 1 (NTX: p = 0.002; BAP: p = 0.0005) and declined, as puberty progressed. A negative correlation was observed between aBMD and biochemical bone markers at the lumbar spine (NTX: r = -0.54, p = 0.05; BAP: r = -0.44, p = 0.01) and in the whole body (NTX: r = -0.60, p = 0.0008; BAP: r = -0.19, p = 0.002). We conclude that the negative relationships between aBMD and biochemical markers suggest a high bone turnover, mainly in prepubertal patients and that the results observed in relation to aBMD and puberty are imputed to the delayed puberty which occurs amongst TS patients. PMID- 14752211 TI - Laparoscopic technique for secondary vaginoplasty in male to female transsexuals using a modified vascularized pedicled sigmoid. AB - Between 1995 and 2002, 53 primary sex reassignments in male to female transsexuals were performed at our division. The objective of this procedure is to mimic the female external and part of the internal genitalia both esthetically and functionally. In 11 of the 53 patients, a secondary vaginal lengthening had to be performed due to a short neovagina. This was achieved using a pedicled sigmoid segment, with an open approach in the first 2 patients and using a laparoscopic method in the following 9. This paper focuses on the laparoscopic technique, its benefits and potential complications. Where a primary vaginoplasty, combining inversion of the penile and scrotal skin flaps, yields unsatisfactory functional results, a secondary vaginoplasty using the pedicled sigmoid represents an elegant means to achieve functional improvement. Furthermore, we report a modified surgical approach to the conventional sigmoid transition. PMID- 14752212 TI - Effects of diazeniumdiolates (NONOates) and methylene blue on the reduction of postoperative adhesion in rats. AB - We investigated the effects of spermine NONOate (SPER/NO), diethylenetriamine NONOate (DETA/NO) and methylene blue for preventing postoperative adhesion in a rat uterine horn model. Before operations, rats were randomly assigned into 6 groups, each composed of 12 rats. These were the sham, control, normal saline, SPER/NO, DETA/NO and methylene blue groups. Each rat was anesthetized with ketamine hydrochloride (40 mg/kg i.v.). The abdominal wall was shaved; the surgical site was scrubbed with povidone iodine and rinsed with sterile saline 3 times before surgery. Under sterile conditions, a 3-cm vertical midline incision was made in all groups. In the sham group, we closed the abdominal wall without any procedure. In the other groups, a 2-cm segment of each uterine horn was injured in 10 spots on the antimesenteric surface using unipolar cautery. Before the final abdominal closure, no adjuvant therapy was administered intraperitoneally to the rats in the control group; 2 ml of normal saline solution, 1% methylene blue solution, SPER/NO (0.5 mg/ml) and DETA/NO (0.1 mg/ml) were instilled into the uterine horns of the rats in the respective groups. The incision was closed with a running 4-0 monofilament delayed absorbable suture in a single layer of muscle and fascia in a running pattern, excluding the peritoneum, and in the covering layer of skin in an interrupted pattern, in all groups. Two weeks after the surgery, all animals were killed, second laparotomies were performed, and the extent and severity of adhesions were determined by a blinded examiner. The adhesion scores of the sham group were significantly lower than those of the other groups (p < 0.05). In the methylene blue, SPER/NO and DETA/NO groups, adhesion scores were significantly lower than in the normal saline and control groups (p < 0.05). However, there were no significant differences related to the extent and severity scores of adhesions between the methylene blue, SPER/NO and DETA/NO groups (p > 0.05). This study showed that SPER/NO, DETA/NO and methylene blue administered at the end of surgery reduced the adhesion formation in a rat uterine horn model. PMID- 14752213 TI - Charcot and art: from a hobby to science. AB - Jean-Martin Charcot not only was one of the founders of modern neurology, but he displayed an exceptionally developed visual perception and memory, with special artistic gifts, which he used first as a hobby and subsequently as a tool in his profession. Previously unpublished drawings emphasize Charcot's talents in caricature, including autoderision. One of the best achievements of Charcot in correlating the clinic with art includes his thorough study of artistic representations of "possessed states", which allowed him to refine his work on hysteria. The artist and the scientist are two unique facets of Charcot, whose permanent coexistence help to understand his legacy. PMID- 14752214 TI - Neurosarcoidosis: clinical experience and diagnostic pitfalls. AB - OBJECTIVE: To describe a group of patients with neurosarcoidosis and to highlight diagnostic difficulties based on current diagnostic criteria. METHODS: The patient database of a general neurological department was searched for patients with established or suspected diagnosis of neurosarcoidosis. Twenty-four patients were identified with definite (n = 3), probable (n = 10) and possible neurosarcoidosis (n = 10). History and clinical, laboratory and imaging data of patients with definite and probable neurosarcoidosis were analyzed. RESULTS: Cranial nerve symptoms were a dominant clinical feature, with the optic nerve being affected most frequently. Cerebrospinal fluid pleocytosis was found in more than half of the patients. Intrathecal IgG synthesis and oligoclonal bands were less frequent. There was a wide array of MRI lesions in both groups. Chest X-ray was false negative in 2 of 5 patients who also underwent a thoracic CT. Therapy with prednisolone was initiated in all patients. After a median of 36 months, 6 of 8 patients with follow-up data of >24 months were still in remission. Aggravation of symptoms required therapy escalation in 2 patients. CONCLUSION: There is a wide range of clinical symptoms and test results in patients with "definite" or "probable" neurosarcoidosis. Because systemic involvement is a crucial diagnostic criterion, extensive medical work-up may be necessary. Prognosis under corticosteroid treatment may be better than previously thought. PMID- 14752215 TI - The first placebo-controlled trial of a special butterbur root extract for the prevention of migraine: reanalysis of efficacy criteria. AB - This is an independent reanalysis of a randomised, placebo-controlled parallel group study on the efficacy and tolerability of a special butterbur root extract (Petadolex) for the prophylaxis of migraine. The original protocol and analysis had a number of major shortcomings. In order to follow regulatory requirements, an independent reanalysis of the original data was performed. Following a 4-week baseline phase, 33 patients were randomised to treatment with two capsules 25 mg butterbur twice a day and 27 to placebo. The mean attack frequency per month decreased from 3.4 at baseline to 1.8 after 3 months (p = 0.0024) in the verum group and from 2.9 to 2.6 in the placebo group (n.s.). The responder rate (improvement of migraine frequency > or =50%) was 45% in the verum group and 15% in the placebo group. Butterbur was well tolerated. This small trial indicates that butterbur may be effective in the prophylaxis of migraine. PMID- 14752216 TI - Diffusion-weighted MRI and 99mTc-HMPAO SPECT in delayed relapsing type of carbon monoxide poisoning: evidence of delayed cytotoxic edema. AB - BACKGROUND: Carbon monoxide (CO) is a common cause of poisoning, and its sequelae include a progressive (25%) and a delayed relapsing form (75%). We report the diffusion-weighted MRI (DWI) findings in the delayed relapsing form of CO poisoning and characterize the types of edema. METHODS: From November 1, 2000 to June 1, 2003, 5 consecutive patients (2 men, 3 women, range of age: 54-67 years), who had the delayed relapsing type of CO poisoning, underwent DWI, conventional MRI, MR angiography and SPECT. CO poisoning was diagnosed by the presence of a typical clinical history, an abnormally increased level of serum carboxyhemoglobin and MRI findings. Apparent diffusion coefficient (ADC) values were measured in all of the abnormal lesions with visual inspection of DWI and T(2)-weighted echo-planar imaging. RESULTS: DWI showed high signal intensities in bilateral periventricular white matter, in the splenium of the corpus callosum, in internal capsules, and brainstem showing moderately decreased ADC values. In the globus pallidus, the ADC values were rather increased with low signal intensities on DWI. Brain SPECT showed decreased perfusion in bilateral white matter and some parts of the cerebral cortex, which correlated well with the DWI findings. CONCLUSIONS: We suggest that prominent, symmetric restricted diffusion can occur in periventricular white matter, brainstem, and corpus callosum after the delayed relapsing type of CO poisoning. Delayed cytotoxic edema can occur in this setting, which provides a new guidance for the pathogenesis of CO poisoning and the differential diagnosis of white matter diseases. PMID- 14752218 TI - Proteome analysis for the identification of tumor-associated biomarkers in gastrointestinal cancer. AB - Gastrointestinal cancers are usually diagnosed at advanced stages, making a curative treatment difficult. Biomarkers can help to overcome this problem by allowing earlier diagnosis, and thus better therapy. Proteomics tools are novel technologies to identify such biomarkers. This review summarizes advances in biomarker detection using two-dimensional gel electrophoresis (2D-PAGE), chromatography and mass spectrometry technologies. 2D-PAGE combined with mass spectrometry has led to the identification of several differentially expressed proteins in cancer tissue. However, for serum analysis, 2D-PAGE has severe limitations. For serum-based cancer diagnosis, surface-enhanced laser desorption ionization time-of-flight (SELDI-TOF) mass spectrometry is a promising new technology. The potential of proteins identified with this technology as novel cancer biomarkers still needs to be confirmed in clinical trials. PMID- 14752219 TI - DNA methylation markers in patients with gastrointestinal cancers. Current understanding, potential applications for disease management and development of diagnostic tools. AB - DNA methylation, the modification of a cytosine nucleotide immediately preceding a guanine base in a stretch of DNA, is rapidly gaining strength in the diagnostic field as a powerful tool to be utilized for the discrimination of neoplastic tissue from its healthy counterpart. This epigenetic modification occurs often in the promoter region of genes and is associated with transcriptional silencing of tumor suppressors or other genes important for normal cellular function. These changes have been found to occur at very early stages in the progression of healthy to malignant phenotype in many cancer types. We are taking a targeted approach to finding methylation-based markers that can be used not only for the early detection of cancer but also for determining risk, monitoring patient response to therapy and even determining the degree of aggressiveness of a tumor. In this paper, we review the progress in our understanding of methylation in gastrointestinal tumors, the potential clinical applications of methylation-based markers and our process for the discovery and validation of highly specific and sensitive markers for the use in these applications. PMID- 14752220 TI - Application of DNA array analyses in the management of gastrointestinal cancer patients. AB - Cancer is a polygenic disease arising from the accumulation of multiple genetic and epigenetic defects in the affected cells. For the majority of cancers, including many malignancies of the gastrointestinal tract, our current means of diagnosis and treatment of the tumors are grossly insufficient. DNA arrays offer the possibility to monitor the expression levels of thousands of mRNA transcripts simultaneously in a single assay, making them ideal tools to study the complex network of transcriptional changes that are associated with the malignant transformation of normal cells. Expression profiling analyses are rapidly expanding our knowledge of tumor cell biology, laying the basis for the development of more sophisticated diagnostic procedures and more effective treatment modalities. This review will discuss current applications and future perspectives of DNA array technology in oncological research and tumor patient management. PMID- 14752221 TI - Identification of colon cancer patients by molecular diagnosis. AB - The important causal role of DNA alterations in cancer development implies that tumor specific DNA mutations might serve as ideal parameters in cancer diagnosis. In particular the detection of mutations in fecal DNA might complement the current methods for the early diagnosis of colorectal tumors. Over the last 10 years many studies have proven the usefulness and the feasibility of the fecal DNA analysis in the diagnosis of colon tumors. Technical difficulties have circumvented the clinical application and the broad screening by molecular diagnosis. Recent methodological proceedings together with the promising outcomes of first multi-target studies have put a diagnostic stool-based DNA test for colorectal tumors within close reach. PMID- 14752222 TI - Molecular detection of circulating cancer cells. Role in diagnosis, prognosis and follow-up of colon cancer patients. AB - BACKGROUND: The circulating tumor cells of solid tumors, especially in colorectal cancer has been of great interest in medical research in the last and new century. METHODS: A literature review of recent data for the preparation of a proposal for clinical applications. RESULTS: Starting with the detection of circulating cancer cells in histological sections, lymph nodes, bone marrow and peripheral blood have recently become the primary samples for these investigations. Following microscopic morphology, new methods were and are being developed continuously including rare cell enrichment and detection techniques like fluorescence microscopy, flow cytometry, RT-PCR and methylated DNA PCR. The sensitivity of these assays reaches down to the concentration of 1 tumor cell/ml blood. A lot of different epithelial markers were targeted with these techniques e.g. cytokeratins, EGFR, CEA, and EMA. Clinically, circulating tumor cells were found as independent prognostic factors in lymph nodes and bone marrow. In blood, their presence appears to be an early marker for recurrence and relapse. The change in their quantitative number is supposed to reflect the chemotherapeutic sensitivity and metastatic growth activity of the tumor. CONCLUSIONS: The detection of circulating tumor cells in bone marrow and lymph nodes is of clinical significance. Their presence in peripheral blood and therapeutic applications need further clarification. PMID- 14752223 TI - Molecular analysis of therapy resistance in gastric cancer. AB - Therapy resistance is the main cause of therapeutic failure and death in patients suffering from gastric carcinoma. Clinical resistance against systemic chemotherapy of gastric cancer is likely to be multifactorial and heterogenous. So far, no significant resistance factor that predicts the clinical outcome of systemic treatment of gastric carcinoma has been identified. In order to gain further understanding of therapy resistance in gastric carcinoma, various in vitro model systems were established. One of these models consists of the parental, drug-sensitive and thermosensitive human gastric carcinoma cell line EPG85-257P, its classical multidrug-resistant variant EPG85-257RDB, its atypical multidrug-resistant subline EPG85-257RNOV and their thermoresistant counterparts EPG85-257P-TR, EPG85-257RDB-TR, and EPG85-257RNOV-TR. This panel of cells was analyzed using morphological, biochemical, cellular and molecular biological methods to identify potential new factors involved in therapy resistance of gastric carcinoma. Cellular alterations that could be identified in these models were evaluated by functional investigations. This review will discuss the current state of knowledge of these new therapy resistance-associated factors, e.g. glypican-3 (GPC3), as well as the impact of well-known drug resistance-associated factors, such as MDR1/P-glycoprotein, on therapy resistance of gastric carcinoma. PMID- 14752224 TI - Alterations of the CARD15/NOD2 gene and the impact on management and treatment of Crohn's disease patients. AB - The recent identification of the CARD15/NOD2 gene as a susceptibility locus for Crohn's disease represents an important step towards the delineation of the immunopathogenesis of inflammatory bowel disease. CARD15 functions as an intracellular receptor for bacterial components and thus represents an important link between inflammatory bowel disease and innate immunity. Three major CARD15/NOD2 gene mutations have been associated with Crohn's disease in Caucasians in several independent studies. Together, they explain about 20% of the genetic susceptibility for Crohn's disease. Genotype-phenotype analyses demonstrated an association of these mutations with ileum-specific disease, an increased incidence of the fibrostenotic phenotype and an earlier age of disease onset. Beside these associations, no other relationship between the CARD15/NOD2 genotype and disease behavior or response to treatment has been detailed so far. Thus, the clinical impact of knowing the patient's genotype is limited at this time. Screening for CARD15 mutations in order to identify high-risk individuals or to introduce an individualized disease management is therefore currently not recommended. PMID- 14752225 TI - Identification of patients with genetic risk factors of pancreatitis: impact on treatment and cancer prevention. AB - Recent data suggest that genetic alterations are relevant risk factors for chronic pancreatitis. The highest risk is associated with autosomal-dominant mutations (N29I, R122H) of the cationic trypsinogen (PRSS1). Further mutations were identified in the genes of the pancreatic trypsin inhibitor (SPINK1) and in the cystic fibrosis transmembrane conductance regulator (CFTR). A remarkable finding was that both molecules were also mutated in patients suffering from alcoholic chronic pancreatitis. According to recent estimations, genetic alterations may be regarded as more severe risk factors than chronic alcohol consumption. To identify patients with mutations, a positive family history could be of help, but mutations were also found in a significant number of those with a negative family history. On the other hand, in approximately 40% of the patients with a positive family history no mutations were found up to now. The age at onset is lower in patients with genetic risk factors; however, no clear limit can be denominated above which a screening is not appropriate. Therefore, in our department genetic screening is offered to all patients with chronic pancreatitis of unclear origin. There is no specific treatment in patients with a genetically based disease. The patients with familial pancreatitis-increased rates of pancreas cancer were described but there is no agreement concerning the prophylactic strategy. Prevention of cancer by routine pancreatectomy, though performed recently, is not justified at the moment. Clinical criteria may be more appropriate to decide the timing and the extent of the operation. PMID- 14752226 TI - Informed consent for molecular-based diagnostic and prognostic studies in the cancer patient. AB - The consent of the subject who is donating cells, tissues, organs or body fluid for research purposes is a precondition for biomedical research on human samples. Before such consent can be obtained, comprehensive information should be provided to the subject by the investigators. This information includes the scope of research, the intended use of subject's body parts and associated data, the risks and benefits associated with the research project, and the right to withdraw their consent at any time without prejudice. Consent is free and must not be obtained under any kind of pressure. The traditional practice of obtaining consent for unspecified future use of biological samples and data generated from clinical trials is no longer adequate for genetic research. However, the investigator has still interest to keep the definition of the field of research as broad as possible. Privacy is an issue of importance for the patient, who is the primary supplier of human samples, and for his relatives. Benefit sharing is another field of contradictory discussion. Normal volunteers are a special case among gene expression studies. An example of informed consent form is provided. PMID- 14752227 TI - Clinical implications of alpha-fetoprotein expression in gastric adenocarcinoma. AB - BACKGROUND: Alpha-fetoprotein (AFP)-producing gastric adenocarcinoma has an extremely poor prognosis. Few cases have been reported in Germany until now. Here we report on a patient with an AFP-producing gastric cancer and a subsequent analysis of AFP expression in a more extended series of gastric cancer patients. MATERIAL AND METHODS: A 62-year-old man was referred to our hospital with suspected gastric cancer. Gastroscopy, ultrasound, and CT scans of the abdomen showed a gastric tumour. Serological and histopathological investigations led to the diagnosis of a poorly differentiated adenocarcinoma expressing AFP. Following diagnosis, a short-term response was achieved by palliative chemotherapy. Expression of AFP and albumin was then investigated on an extended series of 25 patients with gastric cancer and four gastric cancer cell lines using immunohistochemistry and RT-PCR. No patient had suffered from hepatocellular carcinoma or germ cell tumor. RESULTS: Using paraffin-embedded gastric cancer specimens, we found AFP immunohistochemically in 2 of 25 (8%) patients. AFP-mRNA was expressed in 3 (12%) patients and a single gastric cancer cell line (AGS). Albumin-mRNA was not found in any gastric cancer sample or gastric cancer cell line. CONCLUSIONS: Our study shows that AFP-producing gastric adenocarcinomas are an important clinical entity with a frequency that has been underestimated in the past. In the absence of a primary liver tumor, clinicians have to consider a primary gastric cancer, which has a poor prognosis, and merits a more aggressive therapy. PMID- 14752228 TI - Microsatellite alterations in free-circulating serum DNA in patients with ulcerative colitis. AB - BACKGROUND/AIMS: The aim of this study was to find out whether there is free circulating DNA in the serum of patients with ulcerative colitis and whether it is possible to establish a panel of genetic alterations which might be useful markers for diagnostic purposes, staging, or follow up. METHODS: A set of 11 microsatellite markers located on different chromosomes were analyzed in a group of 59 patients with ulcerative colitis. Radiolabeled PCR products from serum and white blood cell DNA were analyzed by autoradiography and correlated with the clinical data from these patients. RESULTS: Seven of 59 patients showed one or more microsatellite alteration(s) in their serum DNA. Six patients had an alteration in one marker, and two patients had changes in 4 markers, respectively. There was no correlation between the frequency of microsatellite alterations and the clinical data. CONCLUSION: From the panel of 11 microsatellite markers used for these studies, 6 of them seem to be well suited for the detection of ulcerative colitis-associated alterations in free circulating serum DNA. In order to increase the specificity of this assay, it is necessary to increase the number of patients to be analyzed and to use a different set of markers. PMID- 14752229 TI - EUK-134 reduces renal dysfunction and injury caused by oxidative and nitrosative stress of the kidney. AB - BACKGROUND/AIMS: Oxidative and nitrosative stress plays important roles in the pathogenesis of renal ischemia/reperfusion (I/R) injury. Here we investigate the effect of EUK-134, a synthetic superoxide dismutase and catalase mimetic, (i) on renal dysfunction and injury caused by I/R in vivo and (ii) on proximal tubular cell (PTC) injury and death caused by oxidative and nitrosative stress. METHODS: Rats, subjected to bilateral renal ischemia (45 min) followed by reperfusion (6 h), were administered EUK-134 (0.3 and 3 mg/kg, i.v.) prior to and during reperfusion, after which biochemical and histological indicators of renal dysfunction and injury were measured. The expression of poly(ADP-ribose) (PAR) and inducible nitric oxide (NO) synthase (iNOS) and nitrotyrosine formation were determined immunohistochemically and used as indicators of oxidative and nitrosative stress. Primary cultures of rat PTCs, isolated and cultured from the kidney cortex, were incubated with hydrogen peroxide (H2O2; 1 mM for 2 h) in the presence of increasing concentrations of EUK-134 (1-100 microM) after which PTC injury and death were measured. The effects of EUK-134 on serum levels of NO in rats subjected to renal I/R or on NO production by PTCs incubated with interferon gamma (IFN-gamma, 100 IU/ml) and bacterial lipopolysaccharide (LPS, 10 microg/ml) in combination for 24 h were also measured. RESULTS: EUK-134 produced a significant reduction in renal dysfunction and injury caused by I/R. Specifically, serum creatinine levels, an indicator of renal dysfunction, were reduced from 227 +/- 11 (n = 12, I/R only) to 146 +/- 9 microM (n = 12, I/R +3 mg/kg EUK-134). Urinary N-acetyl-beta-D-glucosaminidase activity, an indicator of tubular damage, was reduced from 42 +/- 5 (n = 12, I/R only) to 22 +/- 3 IU/l (n = 12, I/R +3 mg/kg EUK-134). EUK-134 significantly reduced renal injury caused by oxidative stress in vivo (reduction in PAR formation), and in vitro EUK-134 reduced PTC injury and death caused by H2O2. However, EUK-134 also reduced nitrosative stress caused by I/R in vivo (reduction of iNOS expression and nitrotyrosine formation), which was reflected by a significant reduction in serum NO levels in rats subjected to renal I/R. Specifically, serum NO levels were reduced from 57 +/- 12 (n = 12, I/R only) to 23 +/- 3 mM (n = 12, I/R +3 mg/kg EUK-134). In vitro, EUK-134 significantly reduced NO production by PTCs incubated with IFN-gamma/LPS. CONCLUSION: We propose that EUK-134 reduces renal I/R injury not only via reduction of oxidative stress, but also by reducing nitrosative stress caused by renal I/R. PMID- 14752230 TI - Irreversible inhibition of [3H]glycine transport into channel catfish erythrocytes by thiol group modifiers. AB - Erythrocytes can take up amino acids from the blood by using a variety of transport systems. GLYT is a key transport protein in the plasma membrane responsible for the Na(2+)-dependent uptake of glycine needed for glutathione biosynthesis. Certain cysteine-specific compounds, particularly mercuric chloride and 5,5'-dithiobis(2-nitrobenzoate), irreversibly inhibited the [(3)H]glycine transport via GLYT by red blood cells isolated from channel catfish. Bimolecular rate constants (k(2)) of 0.556 (mmol/l)(-1) min(-1) and 0.032 (mmol/l)(-1) min( 1), respectively, were calculated for the two inhibitors. Addition of 2 mercaptoethanol 1 min after the initiation of inactivation by mercuric chloride stopped further inactivation, but did not reverse the inhibition. The presence of glycine, but not Na(+) ions, during the preincubation of the cells with each inhibitor markedly reduced the degree of inhibition. Thus cysteinyl residues within the transport protein appear to be vital for the binding and uptake of glycine by channel catfish erythrocytes. PMID- 14752231 TI - Interaction of androgens with cardiotonic drugs in isolated left atrium of rat. AB - Pharmacological concentrations of androgens are known to elicit a rapid positive inotropism in isolated left atrium of male rats. Upon short-term exposure to androgens, an increase in intracellular cAMP levels has been observed, though delayed with respect to the time course of contraction, suggesting that other mechanisms may participate in initiating the contraction. Therefore, the interaction of positive inotropism elicited by ouabain, an inhibitor of Na(+) K(+)-ATPase, and androgens was studied in isolated left atrium of rat. Androgens antagonized ouabain-elicited positive inotropism and increased the basal tone. Vanadate, an inhibitor of the Ca(2+) pump, produced a similar effect as androgens on ouabain-elicited positive inotropism. Therefore, androgens might interact with the Ca(2+) pump and this may explain the increase in basal tone. The conjugation of 5 alpha-dihydrotestosterone with bovine serum albumin produced the same effect, suggesting an extracellular interaction of androgens inhibiting the Na(+) K(+)-ATPase that could increase intracellular Ca(2+) via the Na(+)-Ca(2+) exchange. PMID- 14752232 TI - Effects of HR780, a novel 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor, in Watanabe heritable hyperlipidemic rabbits and cholesterol-fed rabbits. AB - The aim of this study was to evaluate the effects of (+)-(E)-6S-(2-(4-(4 fluorophenyl)-2-(1-methylethyl)-6-phenylpyridin-3-yl)ethenyl)-4R-hydroxy-3,4,5,6 tetrahydro-2H-pyran-2-one (HR780), a novel 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor, on hypercholesterolemia in Watanabe heritable hyperlipidemic (WHHL) rabbits and rabbits fed a diet with 1% cholesterol, in comparison with the effects of simvastatin. Each drug was administered orally to WHHL rabbits for 24 weeks and to 1% cholesterol-fed rabbits for 10 weeks. In WHHL rabbits, HR780 at doses of 1, 2.5 and 5 mg/kg/day reduced the plasma total cholesterol level by 15, 24 and 20%, respectively. In contrast, simvastatin at 5 mg/kg/day lowered the level by 23%. In 1% cholesterol-fed rabbits, HR780 (1 and 2.5 mg/kg/day) was found to inhibit the increases in the plasma total cholesterol and phospholipid levels and liver cholesterol contents in a dose-dependent manner. Simvastatin (5 mg/kg/day) also inhibited their increase. Neither HR780 nor simvastatin increased the contents of cholesterol and total bile acid in the gallbladder bile. In conclusion, long-term treatment with HR780 reduced the plasma cholesterol level in WHHL rabbits and 1% cholesterol-fed rabbits, and decreased the liver cholesterol contents in 1% cholesterol-fed rabbits. PMID- 14752233 TI - 7-OH-DPAT-induced inhibition of norepinephrine release in PC12 cells. AB - The purpose of this study was to investigate mechanisms of suppression of norepinephrine release by 7-OH-DPAT, a dopamine D(2)/D(3) receptor agonist, in PC12 cells pretreated with nerve growth factor (NGF). 7-OH-DPAT caused inhibition of basal and K(+)-evoked norepinephrine release, which could be blocked by pretreatment with raclopride, a D(2)/D(3) receptor antagonist. Moreover, dopamine D(2) and D(3 )receptors were identified by immunocytochemistry. Expression of D(2), D(3), and D(4) mRNAs and their proteins were detected using RT-PCR and immunoblotting. Furthermore, 7-OH-DPAT produced no change in cGMP levels; however, 7-OH-DPAT inhibited forskolin-stimulated cAMP accumulation that was antagonized by pretreatment with raclopride. In addition, 7-OH-DPAT inhibited carbachol-induced Ca(2+) transient, conversely, 7-OH-DPAT had no effect on 4 aminopyridine-induced Ca(2+) transient. Taken together, suppression of cAMP accumulation and calcium mobilization by 7-OH-DPAT is involved in the inhibition of norepinephrine release through activation of dopamine D(2)/D(3) receptors. PMID- 14752234 TI - Anti-hypertension effect of vanylidilol: a phenylaldehyde alpha/beta-adrenoceptor blocker with endothelium-dependent and K+ channels opening-associated vasorelaxant activities. AB - The antihypertensive effect of vanylidilol, a new alpha/beta-adrenoceptor antagonist with endothelium-dependent and K(+)-channel-opening activities, was investigated in normotensive and hypertensive Wistar rats. Vanylidilol competitively antagonized (-)isoproterenol-induced positive chronotropic effects, inotropic effects, and tracheal relaxation effects in isolated rat right atria, left atria, and guinea pig tracheal strips in a concentration-dependent manner. Vanylidilol's apparent pA(2) values were 6.36 +/- 0.08 (right atria), 6.41 +/- 0.07 (left atria), and 6.31 +/- 0.06 (trachea). Vanylidilol also produced a competitive antagonism of phenylephrine-induced contraction in the isolated rat aorta with pA(2) values of 6.79 +/- 0.18. In the radioligand binding assay, vanylidilol inhibited [(3)H]CGP-12177 binding to rat ventricle and lung tissues and [(3)H]prazosin binding to brain membranes with Ki values of 535.17, 2,066.69, and 431.11, respectively. In isolated rat thoracic aorta, vanylidilol's vasorelaxant effects on phenylephrine (10 micromol/l)-induced contractions were attenuated by removing endothelium and by the presence of L-N(G)-nitro arginine methyl ester (L-NAME; 100 micromol/l), methylene blue (10 micromol/l), 1H [1,2,4]oxadiazolol[4,3,-a] quinoxalin-1-one (ODQ; 10 micromol/l), tetraethylammonium (10 mmol/l), glibenclamide (1 micromol/l), apamin (1 micromol/l), and charybdotoxin (0.1 micromol/l). In addition, vanylidilol, in an equally antagonistic activity, inhibited phenylephrine-induced phasic and tonic contractions. Intravenous vanylidilol further reduced mean blood pressure in pentobarbital-anesthetized normotensive Wistar rats in a dose-dependent manner. The oral administration of vanylidilol to conscious spontaneously hypertensive rats had a long-lasting hypotensive effect on the heart rate and decreased it in a dose-dependent manner. Furthermore, vanylidilol's vasodilator effect can be attributed in part to the release of NO or NO-related substance from vascular endothelium, while the endothelium-independent mechanism involved in vanylidilol's relaxation is probably linked to the activation of the K(+) channels and the alpha-adrenoceptor blocking activity in these vessels. PMID- 14752235 TI - Effects of neuropeptides on the sumatriptan-disturbed circulation in the optic nerve head of rabbits. AB - The purpose of this work was to study 'in vivo' the vascular responses of retinal vessels of New Zealand white rabbits to substance P (SP), neurokinin A (NKA), neurokinin B (NKB), senktide, capsaicin (CAPS), and calcitonin gene related peptide (CGRP) before and after selective antagonist administration. We examined the effects of these neuropeptides on the normal circulation in the optic nerve head of the rabbit. Drugs were injected via pars plana through a micropipette system. Ten minutes before perivascular injection of 10 nmol/l sumatriptan (to contract the vessel), a selective antagonist or its solvent was administered. Then, cumulative injection of the agonist was performed. The other eye was used as control. Direct measurement of retinal arteriole diameters was performed using digital angiography. The quantification of the relaxing effect is expressed as percentage related to the precontracted vascular diameter. Microinjection of SP (NK1 receptor agonist) up to 10 nmol/l induced a dose-dependent arteriolar dilating effect [E(max) (mean +/- SEM) 21.3 +/- 2.3%]. After the perivascular preinjection of 1 nmol/l L-668,169 or 1 nmol/l L-733,060 (NK1 receptor antagonists), the SP dose-response curve was shifted to the right. The same results were obtained with NKA (NK2 receptor agonist) which induced the most potent effect of all neuropeptides (E(max) 53.3+/-2.5%). The NK2 receptor antagonists L-659,877 and GR 159897 (1 nmol/l) strongly inhibited this arteriolar vasodilation. As for CGRP, doses up to 10 nmol/l induced a marked vasodilation (E(max) 41.1+/-0.4%) which decreased after microinjection of the selective antagonist CGRP8-37. The NK3 receptor agonists (senktide and NKB) showed a minor vasodilating effect (E(max) 5.1+/-1.2 and 8.0+/-0.9%, respectively). On the contrary, CAPS showed a marked dose-dependent vasodilating effect (E(max) 43.2+/ 2.9%), antagonized by the tachykinin receptor antagonists and CGRP8-37. These results suggest, for the first time, the presence of NK1, NK2, and CGRP receptors on the retinal arteriolar wall of the rabbit. PMID- 14752236 TI - Functional study of the [Ca2+]i signaling pathway in aortas of L-NAME hypertensive rats. AB - A variety of mechanisms has been proposed to suggest that nitric oxide participates in the regulation of smooth muscle free [Ca(2+)](c) (the primary determinant of contractile tone), including inhibition of Ca(2+) influx across the plasma membrane and inhibition of intracellular Ca(2+) release. In view of such considerations, the aim of this study was to investigate the possible alterations in contractile responses induced by drugs that mobilize Ca(2+) from different sources in aortae from N(G)-nitro-L-arginine methyl ester (L-NAME) hypertensive rats (LHR). Treatment with L-NAME did not alter the contractile response induced by phenylephrine; however, indomethacin increased the contraction to phenylephrine only in LHR aortae (1.36 +/- 0.08 g, n = 6, vs. 1.97 +/- 0.09 g, n = 7). Both phenylephrine and caffeine evoked rapid and phasic contractions in intact or denuded aortic rings in Ca(2+)-free solution containing EGTA. Phenylephrine-elicited phasic contractions were lower in normotensive rats (NR; 0.41 +/- 0.05 g, n = 9) than in LHR (0.57 +/- 0.06 g, n = 6) and were increased by endothelium removal only in the NR group (0.64 +/- 0.05 g, n = 6). Conversely, neither with treatment with L-NAME nor endothelium removal altered the phasic contractile responses induced by caffeine. The Ca(2+) influx stimulated with phenylephrine was greater in NR (1.95 +/- 0.08 g; pD(2) 6.06 +/- 0.69; n = 8) than in the LHR denuded aorta (1.63 +/- 0.11 g; pD(2) 3.52 +/- 0.06; n = 6). Similarly, contractions stimulated with phorbol ester in denuded arteries were greater in NR (1.76 +/- 0.08 g, n = 7) than in LHR (1.11 +/- 0.11 g, n = 7). In the same manner, indomethacin failed to alter the contraction stimulated with phorbol ester in NR arteries (2.01 +/- 0.21 g, n = 7), although it completely blocked the inhibitory effect of chronic treatment with L-NAME on this contractile response (1.94 +/- 0.24 g; n = 9). Indomethacin did not change the contractile responses stimulated by increasing concentrations of extracellular Ca(2+) in either NR aortas (1.44 +/- 0.26 g; pD(2) 4.74 +/- 0.79; n = 6) or LHR aorta (1.99 +/- 0.19 g; pD(2) 4.10 +/- 0.47; n = 8). However, in the presence of indomethacin, the Ca(2+) influx was similar in NR and LHR aortae. Taken together, these results suggest that, in this model of hypertension, the increase in agonist-induced release of Ca(2+) from intracellular stores may be partly compensated by inhibition of Ca(2+) influx and that this effect is due to the increased production of the relaxant prostanoid in vascular smooth muscle cells. PMID- 14752237 TI - 4th international symposium on inherited diseases of the pancreas. November 7-9, 2003, Chicago, Ill., USA. PMID- 14752238 TI - Regulation of plasma hemopexin activity by stimulated endothelial or mesangial cells. AB - The pathogenesis of glomerular alterations and proteinuria in corticosteroid responsive nephrotic syndrome (CRNS) is unknown. As an isoform of plasma hemopexin (Hx) with protease activity may be implicated in this disease, we have studied the inhibition of Hx by ADP and reactivation to its active form by endothelial or mesangial cells in vitro. We hypothesized that these cells might potentially be able to convert the inactivated form of Hx (Hxi) to active Hx (Hxa) in vitro, mediated by cellular ecto-ADPase. Since ecto-ADPase (CD39) is upregulated after stimulation of these cells with lipopolysaccharide (LPS) or certain cytokines, we postulated that this conversion might occur specifically after inflammatory stimulation of these cells. Human endothelial or mesangial cell cultures were incubated overnight with or without LPS (10.0 ng/ml) or TNFalpha (10.0 ng/ml), washed and subsequently incubated with Hxi (1.5 mg/ml) in serum-free conditions (Hxi was prepared by treatment of Hxa with ADP or ADP-beta S). After 60 min, supernatants were tested for their capacity to alter glomerular extracellular matrix molecules (i.e. ecto-apyrase) in vitro using standard methods, and compared with Hxi that had not been incubated with cells. Supernatants containing Hxa (1.5 mg/ml) served as positive control. The results show significant activity in supernatants with Hxi (prepared using native ADP). However, Hxi inactivated by ADP-beta-S (which is non-hydrolyzable) could not be reactivated after contact with LPS-stimulated or unstimulated cells in vitro. As ecto-ADPase of these cells is upregulated by LPS, we believe that reactivation of Hxi to Hxa is mediated by cellular ecto-ADPase. Although the relevance of this inflammation-mediated activation mechanism of Hx in patients with CRNS requires further experimentation, our preliminary observations suggesting that this mechanism is corticosteroid dependent may support a potential role of Hxa in CRNS. PMID- 14752239 TI - Low sodium excretion in SIADH patients with low diuresis. AB - It is well known that during low diuresis or low effective circulating volume, salt excretion is low. The aim of this study was to find out whether salt excretion, expressed as either urinary sodium concentration (UNa) or fractional sodium excretion (FENa), and the combined use of FENa and fractional urea excretion (FEurea) still differentiate between hyponatremic SIADH and hyponatremic salt depletion (SD) patients when diuresis is low. The relationships between UNa, FENa and diuresis, indirectly estimated by the urinary to plasma creatinine ratio (U/P creat), were studied in 42 hyponatremic SIADH patients, 21 hyponatremic SD patients and 66 normonatremic controls (CO) of similar age and sex ratio. There was no significant relationship between UNa and U/P creat either in SIADH or in SD or CO patients. FENa and U/P creat were inversely correlated, both in CO (r = -0.72; p < 0.001) and in SIADH (r = -0.68; p < 0.001). SIADH and SD patients can be fairly well differentiated from one another using FENa and U/P creat. Even with high U/P creat values, SIADH patients, despite a sharp decrease in their FENa values, presented still higher FENa values than SD patients did (mean FENa = 0.3 +/- 0.2% in SIADH and 0.1 +/- 0.04% in SD; p < 0.05). However, FENa values of SIADH patients with low diuresis (mean FENa = 0.3 +/- 0.2% for a mean U/P creat = 191 +/- 40) are indistinguishable from those of SD patients with normal urine volumes (mean FENa = 0.2 +/- 0.2% for a mean U/P creat = 92 +/- 30). The combined use of FENa and FEurea remains a reliable way to discriminate SD patients and SIADH patients, as far as the differential limit value for FENa is narrowed to a value of 0.15%, for hyponatremic patients with U/P creat >140. CONCLUSION: In SIADH, FENa values are lower than 0.5%, as soon as U/P creat exceeds a value of 180. In SD patients with U/P creat values exceeding 140, FENa is lower than 0.15% and FEurea lower than 45%. PMID- 14752240 TI - Increased renal fibrosis and expression of renal phosphatidylinositol 4-kinase beta and phospholipase C(gamma1) proteins in piglets exposed to ochratoxin-A. AB - Endemic nephropathy has been linked to exposure of ochratoxin-A (OA) in grains and animal products. The underlying events surrounding this form of renal injury are not well known, partly due to the lack of a suitable animal model of the disease. Therefore, in this study, a pig model of OA-induced renal injury was established and used to examine whether elements of the phosphoinositide signalling pathway are altered in this disease. Weanling piglets were fed diets containing 0, 2, and 4 ppm OA for 6 weeks. Serum creatinine and urea and renal fibrosis were monitored biweekly using serial blood samples and renal biopsies. At termination, the protein levels of renal phosphatidylinositol 4-kinase-beta (PtdIns4Kbeta) and phospholipase C(gamma1) (PLC(gamma1)) were determined using immunoblotting and scanning densitometry. Serum creatinine was elevated by 2 weeks and renal fibrosis was elevated by 4 weeks at both levels of inclusion of OA. At the end of the experimental period, kidney size and water content were elevated, as were the protein levels of renal PtdIns4Kbeta and PLC(gamma1) in OA exposed animals. Therefore, serial biopsies can be used to track changes in renal pathology in the OA-exposed piglet. We conclude that this is a useful model for OA-induced renal injury in which the underlying molecular events associated with this form of renal injury can be studied. PMID- 14752241 TI - A study of VEGF and its receptors in two rat models of proteinuria. AB - BACKGROUND: The high level of expression of vascular endothelial growth factor (VEGF) in normal podocyte foot processes suggests that VEGF has an important role in maintaining normal glomerular function. While altered VEGF expression occurs in many glomerular diseases, a direct role for VEGF in the pathogenesis of proteinuria has not been demonstrated. METHODS: Expression of VEGF and its receptors (VEGFR-1 and VEGFR-2) was examined in passive Heymann nephritis (PHN) and puromycin aminonucleoside nephrosis (PAN), by immunohistochemistry, in situ hybridization, Northern and Western blotting. Inhibition of VEGF in the PAN model was performed by administration of a blocking antibody. RESULTS: In both models, glomeruli showed upregulation of VEGF and VEGF receptors compared to control animals. VEGF mRNA was increased most significantly (5-fold) at day 5 after induction of PHN, prior to the onset of proteinuria, with persistent upregulation (3-fold) at day 21. Increased VEGF mRNA was also seen in PAN, but it was less marked. In situ hybridization and immunohistochemistry localized VEGF predominantly to podocytes. Increased expression of VEGFR-1 and VEGFR-2 protein was seen in glomerular endothelial cells of PHN and PAN rats by immunohistochemistry, as was VEGFR-2 mRNA by in situ hybridization. Upregulation of VEGFR-1 by endothelial cells was more striking in the PAN model than PHN. Administration of a blocking antibody to rats with PAN did not affect proteinuria, creatinine clearance or sodium excretion. CONCLUSION: The expression of VEGF and its receptors is significantly increased in the PHN and PAN rat models of proteinuria suggesting a role for VEGF in the disease process. VEGF may have an important role in promoting glomerular repair in a variety of glomerular diseases. PMID- 14752242 TI - Cytokines and cell adhesion molecules in the inflammatory response during acute pyelonephritis. AB - Urinary tract infection is a common bacterial infection of childhood. Renal parenchymal scarring, a recognised complication of urinary tract infection, is responsible for up to 24% of children entering end-stage renal failure. Why acute inflammation results in renal scarring in some children whilst in others complete resolution occurs without scarring is at present poorly understood. This article reviews the role of the cytokines, adhesion molecules and growth factors in the inflammatory response during acute pyelonephritis and renal parenchymal scarring. We hypothesize that inter-individual variability in cellular response may in part be responsible for this variable clinical outcome. PMID- 14752243 TI - MDR1 haplotypes modify BEN disease risk: a study in Bulgarian patients with Balkan endemic nephropathy compared to healthy controls. AB - BACKGROUND: Balkan endemic nephropathy (BEN) is a slow progressive nephropathy with frequent occurrence of uroepithelial tumors in the upper urinary tract. Genetic factors involved in xenobiotic detoxification mechanisms may cause genetic predisposition to BEN and influence the risk for this disease. Polymorphic MDR1 variants with decreased P-glycoprotein (P-gp) activity modulate the risk for renal neoplasm. We have therefore investigated the impact of MDR1 polymorphisms on BEN manifestation. METHODS: The constitutional genotype frequencies of two SNPs (C3435T and G2677T) in the MDR1 gene in 112 healthy control subjects were investigated and compared with those of 96 patients with BEN. Identification of the SNPs was done with rapid cycle real-time PCR and melting curve analysis with allele-specific probes. RESULTS: The frequency of mutant alleles was comparable in both groups. Significant differences were revealed when the MDR1 haplotypes were analyzed. Individuals with a predicted haplotype 12 (2677G/3435T) were less frequent in BEN cases (frequency 7.3%) than in controls (16.1%, p = 0.006). We found that carriers of the haplotype 12 had a decreased risk for BEN (OR = 0.411; 0.21-0.78). CONCLUSIONS: The data suggest that haplotype 12 is protective against BEN. There is no clear molecular explanation of the MDR1 haplotype effects on the protein activity, which can explain the modified effect of the haplotype 12 on BEN risk. PMID- 14752244 TI - Autoimmune renal injury in C3- and C4-deficient mice: a histological and functional study. AB - BACKGROUND: Complement deficiency predisposes to autoimmune renal disease. Since complement deficient mice are increasingly used to study the immunopathogenesis of renal disease we have determined whether mice deficient in C3 or C4 are susceptible to spontaneous immune-mediated renal injury. METHODS: C3-deficient, C4-deficient and complement-sufficient, wild-type mice were maintained in standard conditions for 1 year at which stage renal function, renal histology, circulating antibody and autoantibody levels were assessed. RESULTS: No significant decline in renal function was demonstrated in the complement deficient mice. However, there was histological evidence of glomerular injury in both the C3- and C4-deficient mice, but of insufficient severity to alter function. Serum IgG2a concentration was significantly lower in C3- and C4 deficient mice. In contrast C3-deficient mice had higher concentrations of serum IgG2b. There was a tendency for mice from all groups, including the complement sufficient mice, to develop autoantibodies. C4-deficient mice had higher titres of anti-dsDNA IgG but otherwise deficient mice had similar autoantibody titres to controls. CONCLUSION: We conclude that C4-deficient mice demonstrate a small increase in autoantibody production at 1 year of age compared to C3-deficient and wild-type mice. Furthermore, although complement-deficient mice exhibit glomerular changes, they are of minor functional significance, and are unlikely to affect the study of experimentally induced renal disease in these mice. PMID- 14752245 TI - Stimulation of NHE3 in OKP cells by an autocrine mechanism. AB - BACKGROUND/AIMS: Chronic hypokalemia increases NHE3 activity in OKP cells. The aim of the present study was to determine whether an autocrine mechanism is involved in this activation. METHODS: After incubation of OKP cells in normal K(+) and low-K(+) media for 24 h, the potassium concentration in the low-K(+) media was adjusted to a normal level. These conditioned media were then used as the normal-K(+) and low-K(+) supernatants. Other OKP cells were incubated in these normal-K(+) and low-K(+) supernatants and the mechanism of Na(+)/H(+) antiporter activation was examined. RESULTS: The EIPA-resistant Na(+)/H(+) antiporter activity of OKP cells increased after 4 h incubation in the low-K(+) supernatant, and the amount of NHE3 protein increased at 24 h. Since both BQ788 and saralasin blocked this antiporter activation, the supernatant concentration of endothelin I (ET-I) and angiotensin II (Ang-II) were measured. The ET-I concentration was reduced, but the Ang-II concentration remained unchanged. There was a significant association between a reduction in the ET-I concentration and an increase in Na(+)/H(+) antiporter activity, but only when Ang-II was present in the supernatant. CONCLUSION: An autocrine mechanism is involved in the activation of NHE3 in OKP cells. Both ET-I and Ang-II play a role in this activation. PMID- 14752246 TI - Reducing dialysate calcium - a reasonable strategy? PMID- 14752247 TI - Effects of reduced dialysate calcium on calcium-phosphorus product and bone metabolism in hemodialysis patients. AB - BACKGROUND: The safety of using reduced calcium dialysate (RDC) in hemodialysis (HD) patients is controversial due to related changes in bone metabolism. In the present study we investigated whether an 18-month treatment period with RDC may induce significant changes in calcium-phosphorus product (CaxP), bone metabolism, and components of the insulin-like growth factor (IGF) system in HD patients. STUDY DESIGN: In this prospective study, 13 HD patients with biochemical signs of diminished or low-normal bone turnover and high CaxP due to high serum calcium level were treated by lowering dialysate calcium from 3.5 to 2.5 mEq/l for 18 months. By specific immunometric assays, serum levels of intact parathyroid hormone (PTH), bone alkaline phosphatase (B-ALP), pyridinoline (PYR), desoxypyridinoline (D-PYR), 25-OH-vitamin D(3) (25-vit D(3)), 1,25-(OH)(2) vitamin D(3) (1,25-vit D(3)), free IGF-I, IGF-II, and IGF-binding protein (IGFBP) 1 to -6 were measured. RESULTS: CaxP decreased significantly from 5.62 (baseline) to 3.95 mmol(2)/l(2) (at 18 months), whereas PTH increased from 81 +/- 57 pg/ml at baseline to 236 +/- 188 at 12 months (p < 0.01), remaining in this range thereafter. Parameters of bone resorption (PYR) as well as formation (B-ALP) significantly increased during RDC, with peak levels after 12 months. Despite increasing doses of oral alfacalcidol, levels of 25-vit D(3) and 1,25-vit D(3) subsequently declined during RDC. In parallel with the changes in bone markers, free IGF-I levels decreased (baseline: 1.9 +/- 0.9 ng/ml, after 18 months: 1.1 +/ 0.7; p < 0.01). The decline of free IGF-I correlated with decreasing levels of IGFBP-3 and increasing levels of IGFBP-1/-4. CONCLUSION: The treatment with RDC effectively lowered CaxP and stimulated bone formation and resorption. The different changes in bone markers and IGF system components mirror the complex effects on bone metabolism. PMID- 14752248 TI - Comorbid depression and platelet serotonin in hemodialysis patients. AB - BACKGROUND/AIM: Comorbid depression often occurs in chronic renal failure patients on hemodialysis (HD). Reduced serotonin (5-HT) function is implicated in the pathophysiology of depression. METHODS: Comorbid depression and different clusters of depressive symptoms were assessed in 79 male HD patients and 35 male depressed psychiatric patients. Platelet 5-HT concentration (a peripheral model for the central serotonergic neurons) was determined in all patients and 80 male healthy controls. RESULTS: Comorbid depression occurred in 50 out of 79 HD patients. Depressed psychiatric patients and depressed HD patients had higher scores of anxiety, retardation, and cognitive symptoms than nondepressed HD patients. Platelet 5-HT concentration was lower in depressed or nondepressed HD patients than in healthy controls, or in depressed patients. Higher platelet 5-HT content was found in depressed psychiatric patients with depressive clusters than in all other patients. Among HD patients, anxious HD patients had a higher platelet 5-HT concentration than HD patients without anxiety symptoms. CONCLUSIONS: Comorbid depression occurred in 63% of HD patients. Dialyzed patients had decreased platelet 5-HT concentration, regardless of the occurrence of comorbid depression. Higher platelet 5-HT concentration was related to anxiety symptoms in HD patients. Our data suggest that platelet 5-HT concentration might be a suitable marker for anxiety symptoms in dialyzed patients. PMID- 14752249 TI - Impaired T cell proliferation and zeta chain phosphorylation after stimulation with staphylococcal enterotoxin-B in hemodialysis patients. AB - BACKGROUND: Patients on regular hemodialysis treatment are in an immunodeficiency state. Several studies have shown defective T cell proliferation after stimulation with various agents. Staphylococcal enterotoxin B (SEB) is a MHC dependent superantigen that triggers proliferation of a large proportion of T cells. T cell activation after stimulation with SEB parallels normal T cell signal transduction. An important and early event in this transduction pathway is the phosphorylation of the zeta chain. In this study, T cell proliferation and zeta chain phosphorylation after stimulation with SEB were evaluated. METHODS: Peripheral blood mononuclear cells (PBMCs) from 24 patients and 14 healthy individuals were isolated and cultured with or without stimulation with SEB (1 ng/ml). Cell proliferation was estimated by immunoenzymatic measurement of bromodeoxyuridine uptake. PBMCs from 8 patients and 6 healthy individuals were isolated and pulsed for 2 min with or without SEB (10 microg/ml). Zeta chain phosphorylation was estimated by immunoprecipitation and immunoblotting with antiphosphotyrosine antibody. RESULTS: Lymphocyte proliferation index after SEB stimulation was lower in hemodialyzed patients. Stimulation of T cells with SEB also resulted in a lower zeta chain phosphorylation in hemodialyzed patients. CONCLUSIONS: Lymphocyte proliferation after MHC-dependent stimulation is impaired in hemodialyzed patients. This proliferation defect is due to impaired zeta chain phosphorylation. PMID- 14752250 TI - Quality of life in male hemodialysis patients. Role of erectile dysfunction. AB - BACKGROUND: Health-related quality of life (QOL) is affected in hemodialysis patients (HD). A number of factors such as age, anemia, and comorbidity had been implicated in decreased QOL. Erectile dysfunction (ED) is a frequent and potentially treatable complication in HD patients. In this cross-sectional study, we aimed to evaluate the possible relation between the QOL and ED in HD patients. PATIENTS AND METHODS: Among the 511 chronic HD patients dialyzed in 11 outpatient HD centers, 148 male patients (mean age: 46 +/- 9 years) were included. The mean time on dialysis was 41 +/- 35 months (range: 3-203 months). Biochemical parameters such as BUN, creatinine, hemoglobin, serum albumin and Kt/V were measured. The QOL of the patients were measured with the short form of Medical Outcomes Study (SF-36), physical component scores (PCS) and mental component scores (MCS) were calculated. The ED was evaluated by the International Index of Erectile Function (IIEF). RESULTS: One hundred and four of the 148 patients (70%) had ED. Hemoglobin levels were correlated with PCS (r = 0.197, p = 0.02) and MCS (r = 0.20, p = 0.019). Patients with ED had lower scores in nearly all the components related to PCS and MCS as compared to patients without ED. IIEF score was correlated with PCS (r = 0.369, p < 0.001) and MCS (r = 0.308, p < 0.001). In linear regression analysis, IIEF score and hemoglobin levels were the independent variables that predicted both PCM and MCS. CONCLUSION: ED, a frequent complication in HD patients, was related to QOL together with anemia. Successful treatment of ED and anemia may lead to improvement in QOL in HD patients. PMID- 14752251 TI - Low initial vitamin B12 levels in Helicobacter pylori--positive patients on chronic hemodialysis. AB - BACKGROUND: Helicobacter pylori has been identified as a possible cause of vitamin B12 deficiency in the general population. We assessed any potential relationship between low cyanocobalamin serum levels and Helicobacter pylori status in hemodialysis patients and subsequently correlated these results with the existence of anemia (a common complication in hemodialysis patients), and macrocytosis. METHODS: In 29 chronic hemodialysis patients, active H. pylori infection was diagnosed using two different methods regardless of digestive symptoms: by searching for bacterial antigens in stools and by the detection of urea breakdown through breath testing. If these results were non-coincident, gastroscopy was performed and antral biopsies obtained. Patients were subsequently divided into group A (H. pylori-positive, n = 8, 28%) and group B (H. pylori-negative, n = 21, 72%). The corresponding initial values of erythrocytic folic acid, vitamin B12 and homocysteine prior to the first hemodialysis session of each patient were retrospectively collected. RESULTS: Vitamin B12 levels (normal 200- 900 pg/ml) were significantly lower in group A compared to group B (225.4 +/- 111.9 vs. 707.9 +/- 258.3 pg/ml, p < 0.011). In group A, 5 patients (63%) had vitamin B12 deficiency (154 +/- 24.6 pg/ml). Baseline hematocrits, erythrocyte folic acid and serum homocysteine levels were not different between the groups, but mean corpuscular volumes were significantly higher in group A compared to group B (109.7 +/-14.1 vs. 91.8 +/- 8.8 fl, p = 0.002). CONCLUSIONS: H. pylori-positive chronic hemodialysis patients may present with lower vitamin B12 blood levels and macrocytosis. H. pylori infection should be suspected in this population when low or low-normal vitamin B12 levels or macrocytosis exist. PMID- 14752252 TI - Effects of simvastatin on plasma lipoproteins and response to arterial injury in wild-type and apolipoprotein-E-deficient mice. AB - OBJECTIVE: To test the non-lipid-lowering effects of simvastatin on the response to injury in normolipidemic and hyperlipidemic mice. METHODS AND RESULTS: Wild type (WT) mice (n = 40) and hyperlipidemic apolipoprotein-E-deficient (apoE(-/-)) mice (n = 40) received normal chow or chow containing simvastatin 100 mg/kg/day prior to bilateral femoral artery wire injury. Intimal hyperplasia and plasma cholesterol concentration were quantified after 4 weeks. Plasma cholesterol in WT mice treated or untreated with simvastatin was similar (100.9 +/- 6.6 vs. 94.3 +/ 17.5 mg/dl). Simvastatin did not affect intimal hyperplasia. In apoE(-/-) mice, intimal hyperplasia was increased 2.3-fold relative to WT mice (17090 +/- 4998 vs. 39490 +/- 16190; p < 0.001). In apoE(-/- )mice, simvastatin caused a paradoxical increase in plasma cholesterol (1094 +/- 60.3 vs. 658 +/- 66.8 mg/dl; p < 0.001), confirmed by FPLC. This was associated with a further increase in intimal area (39490 +/- 16190 vs. 55420 +/- 22590 mm(2); p < 0.01). CONCLUSIONS: (1). Simvastatin had no effect on plasma cholesterol or the response to arterial injury in normolipidemic WT mice; (2). hyperlipidemia was associated with markedly increased intimal hyperplasia, and (3). simvastatin treatment of apoE(-/ ) mice caused paradoxical hyperlipidemia and increased intimal hyperplasia. PMID- 14752253 TI - Expression of costimulatory molecules (4-1BBL and Fas) and major histocompatibility class I chain-related A (MICA) in aortic tissue with Takayasu's arteritis. AB - To further investigate the immunological mechanisms involved, we analyzed the expression of costimulatory molecules in aortic tissue and their counterpart molecules on infiltrating cells of patients with Takayasu's arteritis. We also examined the expression of major histocompatibility complex (MHC) class I chain related (MIC) A in aortic tissue, which is known to be induced by external stress, and its counterpart NKG2D receptors on infiltrating cells. Among these costimulatory molecules, strong expression of 4-1BBL and Fas was induced in the aortic tissue, and most of the infiltrating cells expressed 4-1BB and FasL, suggesting these pathways play critical roles in T-cell-mediated vascular injury. We also found that MICA was strongly induced in the aortic tissue and that at least part of the infiltrating cells expressed NKG2D receptors. Some infiltrating cells - but not vascular smooth muscle cells - seemed to have undergone apoptosis. Our findings strongly suggest that 4-1BB/4-1BBL and Fas/FasL pathways play important roles in vascular injury in Takayasu's arteritis. We assume that gammadelta T cells infiltrated aortic tissue recognizing MICA, resulting in the induction of MHC antigens and costimulatory molecules, and then alphabeta T-cells infiltrated recognizing some auto-antigens presented by MHC antigens, leading to chronic inflammation. PMID- 14752254 TI - Improving the accuracy of NMR structures of large proteins using pseudocontact shifts as long-range restraints. AB - We demonstrate improved accuracy in protein structure determination for large (>/=30 kDa), deuterated proteins (e.g. STAT4(NT)) via the combination of pseudocontact shifts for amide and methyl protons with the available NOEs in methyl-protonated proteins. The improved accuracy is cross validated by Q-factors determined from residual dipolar couplings measured as a result of magnetic susceptibility alignment. The paramagnet is introduced via binding to thiol reactive EDTA, and multiple sites can be serially engineered to obtain data from alternative orientations of the paramagnetic anisotropic susceptibility tensor. The technique is advantageous for systems where the target protein has strong interactions with known alignment media. PMID- 14752255 TI - Probing slow backbone dynamics in proteins using TROSY-based experiments to detect cross-correlated time-modulation of isotropic chemical shifts. AB - The difference in the relaxation rates of zero-quantum (ZQ) and double-quantum (DQ) coherences is the result of three principal mechanisms. These include the cross-correlation between the chemical shift anisotropies of the two participating nuclei, dipolar interactions with remote protons as well as interference effects due to the time-modulation of their isotropic chemical shifts as a consequence of slow micros-ms dynamics. The last effect when present, dominates the others resulting in large differences between the relaxation rates of ZQ and DQ coherences. We present here four sets of TROSY-based (Salzmann et al., 1998) experiments that measure this effect for several pairs of backbone nuclei including (15)N, (13)C(alpha) and (13)C'. These experiments allow the detection of the presence of slow dynamic processes in the protein backbone including correlated motion over two and three bonds. Further, we define a new parameter chi which represents the extent of correlated motion on the slow (micros-ms) timescale. This methodology has been applied to (15)N,(13)C,REDPRO (2)H-labeled (Shekhtman et al., 2002) human ubiquitin. The ubiquitin backbone is seen to exhibit extensive dynamics on the slow timescale. This is most pronounced in several residues in the N-terminal region of the alpha-helix and in the loop connecting the strands beta(4) and beta(5). These residues which include Glu24, Asn25, Glu51 and Asp 52 form a continuous surface. As an additional benefit, the measured rates confirm the dependence of the (13)C(alpha) chemical shift tensor on local secondary structure of the protein backbone. PMID- 14752256 TI - RFDR with adiabatic inversion pulses: application to internuclear distance measurements. AB - In the context of the structural characterisation of biomolecular systems via MAS solid state NMR, the potential utility of homonuclear dipolar recoupling with adiabatic inversion pulses has been assessed via numerical simulations and experimental measurements. The results obtained suggest that it is possible to obtain reliable estimates of internuclear distances via an analysis of the initial cross-peak intensity buildup curves generated from two-dimensional adiabatic inversion pulse driven longitudinal magnetisation exchange experiments. PMID- 14752257 TI - Protein signal assignments using specific labeling and cell-free synthesis. AB - The goal of structural genomics initiatives is to determine complete sets of protein structures that represent recently sequenced genomes. The development of new high throughput methods is an essential aspect of this enterprise. Residue type and sequential assignments obtained from specifically labeled samples, when combined with 3D heteronuclear data, can significantly increase the efficiency and accuracy of the assignment process, the first step in structure determination by NMR. A protocol for the design of specifically labeled samples with high information content is presented along with a description of the experiments used to extract essential information using 2D versions of 3D heteronuclear experiments. In vitro protein synthesis methods were used to produce four specifically labeled samples of the 23.5 kDa protein phosphoserine phosphatase (PSP) from Methanoccous jannaschii (MJ1594). Each sample contained two (13)C/(15)N-labeled amino acids and one (15)N-labeled amino acid. The 135 type and 14 sequential assignments obtained from these samples were used in conjunction with 3D data obtained from uniformly (13)C/(15)N-labeled and (2)H/(13)C/(15)N-labeled protein to manually assign the backbone (1)H(N), (15)N, (13)CO, (13)C(alpha), and (13)C(beta) signals. Using an automated assignment algorithm, 30% more assignments were obtained when the type and sequential assignments were used in the calculations. PMID- 14752258 TI - Paramagnetism-based restraints for Xplor-NIH. AB - Modules that use paramagnetism-based NMR restraints have been developed and integrated in the well known program for solution structure determination Xplor NIH; the complete set of such modules is called PARArestraints for Xplor-NIH. Paramagnetism-based restraints are paramagnetic relaxation enhancements, pseudocontact shifts, residual dipolar couplings due to metal and overall magnetic anisotropy, and cross correlation between Curie relaxation and nuclear nuclear dipolar relaxation. The complete program has been tested by back calculating NOEs and paramagnetism-based restraints from the X-ray structure of cytochrome c (553) from B. pasteurii. Furthermore, the same experimental restraints previously used to determine the solution structure of cytochrome c (553) itself, of cytochrome b (5), and of calbindin D(9k) with the program PARAMAGNETIC DYANA, have been used for structure calculations by using PARArestraints for Xplor-NIH. The agreement between the two programs is quite satisfactory and validates both protocols. PMID- 14752259 TI - Triple quantum decoherence under multiple refocusing: slow correlated chemical shift modulations of C' and N nuclei in proteins. AB - A new experiment allows the identification of residues that feature slow conformational exchange in macromolecules. Rotations about dihedral angles that are slower than the global correlation time tau(c) cause a modulation of the isotropic chemical shifts of the nuclei. If these fluctuations are correlated they induce a differential line broadening between three-spin single-quantum and triple-quantum coherences involving three nuclei such as the carbonyl C', the neighbouring amide nitrogen N and the amide proton H(N) belonging to a pair of consecutive amino acids. A cross-correlated relaxation rate R (CS/CS)(C'N) can be determined that corresponds to the sum of the isotropic and anisotropic contributions to the chemical shift modulations of the carbonyl carbon and nitrogen nuclei. Only the isotropic contributions depend on the pulse repetition rate of a multiple-refocusing sequence. An attenuation of the relaxation rate with increasing pulse repetition rate can therefore be attributed to slow motions. The asparagine N25 residue of ubiquitin, located in the first alpha helix, is shown to feature significant slow conformational exchange. PMID- 14752260 TI - Application of correlated residual dipolar couplings to the determination of the molecular alignment tensor magnitude of oriented proteins and nucleic acids. AB - Residual dipolar couplings (RDC) between nuclear spins in partially aligned samples offer unique insights into biomacromolecular structure and dynamics. To fully benefit from the RDC data, accurate knowledge of the magnitude ( D (a)) and rhombicity ( R ) of the molecular alignment tensor, A, is important. An extended histogram method (EHM) is presented which extracts these parameters more effectively from dipolar coupling data. The method exploits the correlated nature of RDCs for structural elements of planar geometry, such as the one-bond (13)C'(i)-(13)C(i)(alpha), (13)C'(i)-(15)N(i+1), and (15)N(i+1)-(1)H(N)(i+1) couplings in peptide bonds of proteins, or suitably chosen combinations of (1) D (C1'H1'), (1) D (C2'H2'), (1) D (C1'C2'), (2) D (C2'H1'), (2) D (C1'H2'), and (3) D (H1'H2') couplings in nucleic acids, to generate an arbitrarily large number of synthetic RDCs. These synthetic couplings result in substantially improved histograms and resulting values of D (a) and R, compared with histograms generated solely from the original sets of correlated RDCs, particularly when the number of planar fragments for which couplings are available is small. An alternative method, complementary to the EHM, is also described, which uses a systematic grid search procedure, based on least-squares fitting of sets of correlated RDCs to structural elements of known geometry, and provides an unambiguous lower limit for the degree of molecular alignment. PMID- 14752261 TI - 3D TROSY-HNCA(coded)CB and TROSY-HNCA(coded)CO experiments: triple resonance NMR experiments with two sequential connectivity pathways and high sensitivity. AB - The concept of chemical shift-coding monitors chemical shifts in multi dimensional NMR experiments without additional polarization transfer elements and evolution periods. The chemical shifts are coded in the line-shape of the cross peak through an apparent scalar coupling dependent upon the chemical shift. This concept is applied to the three-dimensional triple-resonance experiment HNCA adding the information of (13)C(beta) or (13)C' chemical shifts. On average, the proposed TROSY-HNCA(coded)CB experiment is a factor of 2 less sensitive than the HNCA experiment. However, it contains correlations via the chemical shifts of both (13)C(alpha) and (13)C(beta), and provides up to three times higher resolution along the (13)C(alpha) chemical shift axis. Thus, it dramatically reduces ambiguities in linking the spin systems of adjacent residues in the protein sequence during the sequential assignment. The TROSY-HNCA(coded)CO experiment which is conceptually similar contains correlations via the chemical shifts of (13)C(alpha) and (13)C' without major signal losses. The proposed triple resonance experiments are applied to a approximately 70% (2)H, approximately 85% (13)C,(15)N labeled protein with a molecular weight of 25 kDa. PMID- 14752262 TI - 1H, 15N and 13C NMR resonance assignments of staphostatin A, a specific Staphylococcus aureus cysteine proteinase inhibitor. AB - The increasing antibiotic resistance of an important human pathogen Staphylococcus aureus calls for the development of new therapeutic strategies. Staphylococcal cysteine proteases have been suggested as targets for such therapies. The recent discovery of staphostatins, specific protein inhibitors of these enzymes, gives prospects for the design and production of synthetic, low molecular weight analogs which might become drugs. We have decided to structurally characterize staphostatin A, a representative inhibitor of staphylococcal cysteine proteases, and to assess its binding mode to the target protease with the view of clarifying the specificity determinants. Here we report the (1)H, (15)N and (13)C NMR resonance assignments of staphostatin A. PMID- 14752264 TI - Resonance assignments and topology of the 15N, 13C labelled 23 kDa core domain of xenopus Mob1. PMID- 14752263 TI - Complete 1H, 15N and 13C assignment of the soluble domain of the ba3 oxidase subunit II of Thermus thermophilus in the reduced state. PMID- 14752265 TI - 1H, 13C, and 15N backbone assignments and secondary structure for the 60.8 kD dimer of the NAD+ synthetase from Bacillus subtilis. PMID- 14752266 TI - 1H, 13C and 15N resonance assignments of the region 1463-1617 of the mouse p53 binding protein 1 (53BP1). PMID- 14752267 TI - Sequential 1H, 15N and 13C NMR assignment of human calbindin D28k. PMID- 14752268 TI - Backbone 1H, 13C and 15N resonance assignments for the 18.7 kDa GTPase domain of Escherichia coli MnmE protein. PMID- 14752270 TI - Guarding the Bcl-2 army. PMID- 14752269 TI - Complete backbone resonance assignments of p47: the 41kDa adaptor protein of the AAA ATPase p97. PMID- 14752271 TI - Smurf1: a link between cell polarity and ubiquitination. AB - Members of the Rho family of small guanosine triphosphatases are well known for their important functions in the dynamic regulation of actin cytoskeleton. We recently found that a HECT domain E3 ubiquitin ligase, called Smurf1, regulates cell polarity and protrusion formation by targeting RhoA for degradation at cellular protrusions. Smurf1 regulates these functions as a partner of protein kinase Cxi, a component of the polarity complex. Furthermore, using siRNA mediated knockdown, we demonstrated this pathway is required to maintain the transformed morphology and motility of a tumor cell. Smurf1 thus provides a link between the control of cell polarity and ubiquitin-mediated RhoA degradation during directional cell movements. Here we further discuss the mechanism by which the spatial control of Smurf1 activity is accomplished and the potential implications of these findings in cancer and development. PMID- 14752272 TI - Polycomb group proteins in cell cycle progression and cancer. AB - Epigenetic deregulation of gene expression is emerging as key mechanism in tumorigenesis. Deregulated activity of the chromatin remodeling Polycomb Repressive Complex 2 (PRC2) has recently been shown to be a frequent event in human tumors. Here we discuss these findings and speculate on the role of the PRC2 complex in controlling gene expression during normal cellular proliferation and cancer development. PMID- 14752273 TI - Cdk1: unsung hero of S phase? AB - Cdk2 has been viewed as a key cell cycle regulator that is essential for S phase progression. The recent discovery that Cdk2 is not required for cell proliferation in mice now shows that other factors must be able to replace Cdk2 in stimulating DNA replication. Experiments performed in Xenopus egg extracts identify the mitotic protein kinases Cdk1/Cyclin B and Cdk1/Cyclin A as likely candidates. These observations raise the intriguing possibility that Cdk1 normally participates in genome duplication in wild type cells. PMID- 14752274 TI - Neosis and its potential role in cancer development and chemoresistance. PMID- 14752275 TI - Proteomic analysis of kinase inhibitor selectivity and function. AB - Small molecule inhibitors of protein kinases have become highly popular tools in signal transduction research, despite the fact that rather limited data about their respective selectivities have been available. We established an efficient chemical proteomics method to characterize the cellular targets of the widely used inhibitor SB 203580, which was deemed to be rather specific for p38 kinase. Our results revealed several protein kinases as high affinity targets of SB 203580 and therefore imply a far more complicated cellular mode of action of this inhibitor than previously assumed. This raises the important question whether a lack of selectivity is inherent to many other "specific" inhibitors of protein kinases and warrants their evaluation employing experimental approaches adapted from our described proteomic technique. PMID- 14752276 TI - Hierarchical order of phosphorylation events commits Cdc25A to betaTrCP-dependent degradation. AB - We have recently demonstrated that regulation of Cdc25A protein abundance during S phase and in response to DNA damage is mediated by SCF(betaTrCP) activity. Based on sequence homology of known betaTrCP substrates, we found that Cdc25A contains a conserved motif (DSG), phosphorylation of which is required for interaction with betaTrCP.1 Here, we show that phosphorylation at Ser 82 within the DSG motif anchors Cdc25A to betaTrCP and that Chk1-dependent phosphorylation at Ser 76 affects this interaction as well as betaTrCP-dependent degradation. We propose that a hierarchical order of phosphorylation events commits Cdc25A to betaTrCP-dependent degradation. According to our model, phosphorylation at Ser 76 is a "priming" step required for Ser 82 phosphorylation, which in turn allows recruitment of Cdc25A by betaTrCP and subsequent betaTrCP-dependent degradation. PMID- 14752277 TI - The cyclin D3 knockout: a pound of redundancy with a dash of tissue specificity. AB - Progression through each phase of the cell division cycle is driven by specific cyclin/CDK holoenzymes complexes, each complex having a discrete subset of critical substrates. Phosphorylation of these substrates by the appropriate cyclin/CDK complex in turn directs regulated cell cycle progression. Based on this notion, a reasonable assumption is that elimination of any one of these complexes would be catastrophic with regard to cellular and organismal development. Over the past year however, we have witnessed elegant work that challenges the notion that cyclin/CDK holoenzymes target unique and specific substrates. Indeed in the most recent work, Sicinska et al. report that targeted deletion of the G1 cyclin, cyclin D3, disrupts proliferation of immature T lymphocytes, but does not provide a function essential for organismal development or survival. PMID- 14752278 TI - Linking TGF-beta-mediated Cdc25A inhibition and cytoskeletal regulation through RhoA/p160(ROCK) signaling. AB - Transforming growth factor-beta (TGF-beta) can mediate G(1)/S cell-cycle inhibition and changes in the cytoskeletal organization through multiple parallel downstream signaling pathways. Recent findings regarding TGF-beta-mediated cell cycle checkpoint control and epithelial to mesenchymal transition have converged to the RhoA/p160(ROCK) signaling pathway. The activation of TGF-beta-mediated p160(ROCK)rapidly inhibits the Cdc25A phosphatase as a component of the G(1)/S checkpoint control at the time cytoskeletal re-organization occurs. This can be likened to the ability to preserve genomic integrity in circumstances of genotoxic stress. The inactivation of the RhoA/p160(ROCK) pathway may be a mechanism by which cancer cells bypass growth inhibition even in the presence of TGF-beta. PMID- 14752279 TI - Determinants for Aurora-A activation and Aurora-B discrimination by TPX2. AB - The mitotic kinases Aurora-A and Aurora-B have similar amino-acid sequences but are differently localised and regulated during cell division. The basis for their interactions with different and specific regulators is unclear. Surprisingly, our recent structural studies indicate that TPX2 regulates Aurora-A activity by binding at a site that is conserved almost completely on Aurora-B. Here we investigate molecular determinants of TPX2-Aurora-A recognition. Using structure based mutagenesis, we show that a single amino-acid difference on the surface of the kinase catalytic domain is key to the precision with which TPX2 discriminates between Aurora-A and Aurora-B. The conservation at this amino-acid position suggests that this discriminatory mechanism is likely to be conserved in higher eukaryotes. PMID- 14752280 TI - Increased levels of serum sulfite in patients with acute pneumonia. AB - Sulfite, a common air pollutant, is toxic for humans, causing hypersensitivity or chronic airway diseases. We previously reported that sulfite is actively produced from neutrophils by stimulation with bacterial endotoxin, lipopolysaccharide (LPS). We also found that the serum sulfite concentration is increased in a rat model of sepsis induced by systemic injection of LPS. However, information on sulfite metabolism in human inflammatory conditions is limited. In the current study, the serum concentration of sulfite was determined in 25 patients with acute pneumonia. Serum sulfite concentration in pneumonia patients was significantly higher than that in control subjects (3.75 +/- 0.88 vs. 1.23 +/- 0.48 microM, respectively, P < 0.05). Among 20 patients, serum sulfite was serially determined before and after antibiotic therapy. The levels of serum sulfite were significantly reduced during the recovery phase compared with those during the acute phase (1.34 +/- 0.56 vs. 3.65 +/- 0.92 microM, respectively, P < 0.05). Moreover, neutrophils obtained from three patients during the acute phase of pneumonia spontaneously produced higher amounts of sulfite in vitro than those obtained after recovery. There was a close positive correlation (r = 0.71, P < 0.05) between serum sulfite and C-reactive protein (CRP) in patients with pneumonia. Taken together, the current findings suggest that serum sulfite increases during systemic inflammation in humans. Activated neutrophils might be responsible, at least in part, for the up-regulation of sulfite. Given various biological effects reported previously, sulfite may act as a mediator in inflammation. PMID- 14752281 TI - E. Coli sepsis induces profound mechanoenergetic inefficiency in the porcine left ventricle. AB - Myocardial dysfunction is believed to be a central part of septic multiorgan manifestations. The aim of the present study was to assess whether E. coli sepsis in an in vivo model would induce a dysfunction in the relationship between mechanical work and energy consumption in the left ventricle (LV). Accordingly, we measured hemodynamics, left ventricular pressure-volume area (PVA), and myocardial oxygen consumption (MVo2) in deeply anesthetized pigs. Eight pigs received 2.0 +/- 0.5 x 10(9) E. coli bacteria intravenously, and seven served as controls. Compared with baseline and the control group, no alternations were observed in LV diastolic function or indices of contractility in the septic group. The MVo2-PVA relationship was highly linear in both groups (all r2 = 0.96 0.99). At 5 h, the y-axis intercept of the MVo2-PVA relationship (nonmechanical MVo2) had increased in the sepsis group by 70% compared with baseline (P = 0.004) and by 60% compared with the control group (P = 0.003). Contractile efficiency (the inverse of the MVo2-PVA slope) remained unchanged over time and between groups. The study demonstrates a profound increase in nonmechanical oxygen consumption during E. coli sepsis in the LV. PMID- 14752282 TI - Competitive and noncompetitive inhibition of myocardial cytochrome C oxidase in sepsis. AB - Sepsis is the most common cause of death in intensive care units worldwide. The basic pathophysiologic defect in sepsis, causing functional abnormalities in many organ systems, remains elusive. One potential cause is disruption of oxidative phosphorylation in mitochondria. Here, we report that oxidation of cytochrome c by myocardial cytochrome c oxidase, the terminal oxidase in the electron transport chain, is competitively inhibited early in experimental sepsis (cecal ligation with single or double 23-gauge puncture) in mice. In severe sepsis (cecal ligation and double puncture, 75% mortality at 48 h), inhibition becomes noncompetitive by 48 h. The development of noncompetitive inhibition is associated with a decrease in heme a,a3 content, which is the key active site in the functional subunit (I) and catalyzes the reduction of molecular oxygen. In addition, there are persistently decreased steady-state levels of subunit I mRNA and protein after cecal ligation and double puncture. Both loss of heme and loss of subunit I could explain the observed irreversible inhibition of cytochrome c oxidase. Noncompetitive inhibition of cytochrome c oxidase may interrupt oxidative phosphorylation, leading to sepsis-associated cardiac depression. Importantly, this abnormality may underlie sepsis-associated dysfunction in other organ systems. PMID- 14752284 TI - Antibiotics improve survival in sepsis independent of injury severity but do not change mortality in mice with markedly elevated interleukin 6 levels. AB - Genetically identical mice have a heterogeneous response to antibiotic therapy in sepsis, with only a subset deriving therapeutic benefit. We sought to determine whether the severity of a septic insult correlates with the survival benefit conferred by antibiotics. We also sought to determine whether antibiotics given 12 h after injury alter survival in animals predicted to die based upon high interleukin (IL)-6 levels drawn 6 h earlier. Adult male ND4 mice (n = 363) were subjected to double-puncture cecal ligation and puncture (CLP) with a 19-, 21-, or 23-gauge needle. Animals were randomized to receive imipenem or 0.9% NaCl every 12 h after CLP for 5 days. Ten-day survival was 16%, 26%, and 52%, respectively, for untreated animals. Antibiotics decreased the absolute risk of death 17% to 23% regardless of injury severity. In a separate cohort, mice (n = 37) were subjected to single or double-puncture CLP with a 21-gauge needle. IL-6 levels were assayed 6 h postoperatively and mice were followed for survival. Levels greater than 14,000 pg/mL were identified as predicting a 100% mortality (7/7 animals dead). A third set of mice (n = 94) then underwent double-puncture CLP with either 21-, 23-, or 25-gauge needle and had IL-6 levels measured in a similar fashion. Animals were randomized to receive imipenem or 0.9% NaCl beginning 12 h postoperatively (6 h after IL-6 levels were drawn) and continued for 5 days or until death. Although antibiotics decreased mortality overall, all animals with IL-6 levels greater than 14,000 pg/mL (n = 13) died, regardless of whether they received antibiotics or the gauge of needle used. These results indicate that antibiotics improve outcome in murine sepsis, regardless of injury severity. Furthermore, there is a threshold IL-6 level that can be identified 6 h after sepsis above which animals are destined to die, and antibiotic treatment does not alter their outcome. PMID- 14752283 TI - Antibiotic treatment in a murine model of sepsis: impact on cytokines and endotoxin release. AB - Experimental and clinical studies in sepsis indicate that antibiotic therapy may induce the release of endotoxin (LPS) from the outer membrane of gram-negative bacteria and therefore may affect the physiologic response and survival. The aim of this study was to evaluate if antibiotics commonly used to treat secondary peritonitis are capable of changing survival rates, proinflammatory and anti inflammatory cytokine concentrations, and the release of endotoxin in a murine model of sepsis. Sepsis was induced by cecal ligation and puncture (CLP) in Swiss mice using an 18-gauge needle. The animals received injections of saline solution or imipenem or a combination of ciprofloxacin plus clindamycin every 8 h for 3 days. Antibiotic treatment induced an increase in survival rate and decreased plasma and peritoneal fluid levels of TNF-alpha and IL-6 at 6 and 24 h after CLP as compared with saline-treated animals. Antibiotic-treated animals also showed an early (6 h) decrease and a late (24 h) increase in IL-10 concentration in the peritoneal fluid. LPS concentrations were elevated in all groups, but imipenem treated animals showed higher levels (2.2 EU/mL) than ciprofloxacin plus clindamycin (1.3 EU/mL) and saline-treated (1.5 EU/mL) groups. We conclude that antibiotic-induced endotoxin release is not a major determinant in the inflammatory response and prognosis in murine models of sepsis. PMID- 14752285 TI - Inhibition of poly (ADP-ribose) polymerase attenuates acute lung injury in an ovine model of sepsis. AB - It is known that in various pathophysiological conditions, reactive oxidants cause DNA strand breakage and subsequent activation of the nuclear enzyme poly(ADP ribose) polymerase (PARP). Activation of PARP results in cellular dysfunction. We hypothesized that pharmacological inhibition of PARP reduces the damage in the ovine model of acute lung injury (ALI). After smoke inhalation, Pseudomonas aeruginosa (5 x 109 cfu/kg) was instilled into both lungs. All of the animals were mechanically ventilated with 100% O2. The infusion of the PARP inhibitor (INO-1001, n = 6) began 1 h after the injury and thereafter through 24 h (3 mg bolus + 0.3 mg/kg/h, i.v.). Control animals (n = 6) were treated with saline. Sham injury animals (n = 8) received sham smoke and were mechanically ventilated in the same fashion. One-half of those sham animals (n = 4) were given the same dose of INO-1001. PaO2/FiO2 ratio at 24 h in saline and in the INO-1001 treated groups were 95 +/- 22 and 181 +/- 22, respectively (P < 0.05). Peak airway pressure at 24 h in the saline- and INO-1001-treated groups was 32.6 +/- 3.0 and 24.4 +/- 2.2, respectively (P < 0.05). Pulmonary shunt fraction was also significantly attenuated. INO-1001 treatment reduced pulmonary histological injury and attenuated poly (ADP-ribose) accumulation in the lung. In conclusion, inhibition of PARP improved the ALI after smoke inhalation and pneumonia. The results suggest that the activation of PARP plays a role in the pathophysiology of ALI in sheep. PMID- 14752286 TI - A protective role of interleukin 11 on hepatic injury in acute endotoxemia. AB - The liver is one of the major target organs affected in sepsis, and its failure always results in critical consequences. It has been reported that recombinant human interleukin 11 (rhIL-11), a pleiotropic cytokine, may be useful in the treatment of sepsis. However, the effect of IL-11 specifically on the hepatic failure in sepsis has not been evaluated. In the present study, we examined the effect of rhIL-11 on the hepatic injury in a rat endotoxemia model. Acute endotoxemia was induced in male Sprague-Dawley rats by intraperitoneal injection (i.p.) of bacterial lipopolysaccharide (LPS, 20 mg/kg). Immediately after injection of LPS, rats were treated with rhIL-11 (150 microg/kg, i.p.) or the vehicle. LPS treatment induced severe hepatic injury as revealed by marked increases in serum alanine transaminase (ALT) and aspartate transaminase (AST) activities, extensive hepatocyte necrosis, tumor necrosis factor-alpha (TNF alpha) mRNA, inducible nitric oxide synthase (iNOS) mRNA, and DNA-binding activity of nuclear factor-kappaB (NF-kappaB). In contrast, rhIL-11 treatment significantly ameliorated the LPS-induced hepatic injury, as judged by marked improvement in all these indices. In addition, rhIL-11 treatment markedly decreased LPS-induced mortality. These results indicate that rhIL-11 plays a significant protective role in LPS-induced hepatic injury in acute endotoxemia. PMID- 14752287 TI - Antithrombin III diminishes production of oxygen radical in endotoxin-infused rat lung. AB - The interaction of antithrombin III (AT) with cell surface glycosaminoglycans is known to have an inhibitory effect on inflammatory processes. We evaluated the effect of AT on endotoxin-induced production of oxygen radical in the pulmonary circulation using a fluorescent imaging technique. Also measured was the myeloperoxidase content of the lung, which served as an index of neutrophil accumulation, and neutrophil F-actin levels. Four groups of rats were infused for 2 h with endotoxin at 4.5 mg/kg/h (Et group), physiological saline (CT group), 100 U/kg of AT + endotoxin (AT group), or 100 U/kg of low-heparin-affinity latent AT + endotoxin (L-AT group), respectively. Production of oxygen radical, neutrophil accumulation, and neutrophil F-actin levels were all significantly higher in the ET and L-AT groups than in the CT or AT group. Moreover; the levels of myeloperoxidase within the lung were well correlated with levels of oxygen radical production, which was consistent with the electron microscopic finding that cerium was deposited almost exclusively around neutrophils. Thus, it appears that AT most likely reduces F-actin formation in neutrophil by binding to glycosaminoglycans (e.g., syndecan-4) on the neutrophil, thereby reducing neutrophil accumulation in the lung, which would in turn inhibit oxygen radical production in the lung. PMID- 14752288 TI - Involvement of CD14 and toll-like receptor 4 in the acute phase response of serum amyloid A proteins and serum amyloid P component in the liver after burn injury. AB - Acute phase proteins such as serum amyloid A proteins (SAAs) and serum amyloid P component (SAP) are induced in the liver after various insults (e.g., infection, injury). The cellular and molecular mechanisms controlling the expression of these acute phase proteins may be specifically designed for different insults. The roles of two central molecules of the lipopolysaccharide (LPS)-mediated inflammation pathway (CD14 and toll-like receptor 4 [Tlr4]) were investigated for the regulation of SAAs and SAP in the liver of mice after an 18% total body surface area burn injury. RT-PCR analysis revealed a subtype- and time-dependent induction of SAA mRNAs between 3 h and 3 days, while there was a peak induction of SAP mRNA at day 1. Marked elevations of SAA and SAP protein levels at day 1 supported the mRNA data. Furthermore, a differential regulation of SAAs and SAP mRNAs was noted between CD14 knockout (KO) and their control mice after injury. SAA protein was induced to a lesser degree after injury in C3H/HeJ (Tlr4 defective) mice than in their control mice. In addition, in both CD14 KO and C3H/HeJ mice, the induction of SAP protein was significantly reduced compared with respective controls. These data provide evidence that CD14 and Tlr4 participate, at least in part, in a cascade of signaling events that control the immediate-early and differential induction of SAAs and SAP in the liver after injury. They also suggest that LPS may be one of the initial inducing agents associated with these acute phase responses in the liver after injury. PMID- 14752289 TI - Effects of prolactin level on burn-induced aberrations in myelopoiesis. AB - In this study, we sought to determine if prolactin (PRL) had any influence on burn-induced alterations in myelopoiesis and serum IL-6, IL-10, IL-12, IFN-gamma, TNF-alpha, and MCP-1 levels. To do this, we used mice that were PRL normal, PRL deficient, or hyperprolactinemic and had received a 15% total body surface area burn, sham treatment, or no treatment. We performed clonogenic assays of bone marrow cells, and we found that sham treatment significantly decreased monocyte/macrophage (M) colony formation relative to the control group in the PRL deficient and PRL-normal mice (P < 0.01). Hyperprolactinemia attenuated the sham induced decrease in M colony formation. Burn injury significantly increased M colony formation relative to the sham group with an equal significance in the PRL deficient and PRL-normal mice (P < 0.05). We also showed that burn led to a significant increase in GM colony formation relative to the sham group. This burn induced increase was significant in the PRL-normal (P < 0.05) and the PRL deficient (P < 0.01) mice. In the PRL-normal mice, burn injury caused a 2.1-fold increase in the GM colony number, whereas in the PRL-deficient mice burn led to a 2.6-fold increase in GM colony number. When comparing the effects of burn injury on colony formation to the control groups, there were no significant differences seen, irrespective of the PRL level. We observed that all of the cytokines studied, with the exception of IL-10, were influenced by either sham treatment, burn injury, or both forms of stress. This stress-induced response occurred most often in animals that were either hypo- or hyperprolactinemic. We conclude that the PRL level was able to influence the sham-induced and burn-induced alterations in GM and M colony formation. Under euprolactinemic conditions, mice exhibited less often with stress-induced serum cytokine level alterations. We did not find any significant correlations with any of the serum cytokine levels and the ability to form colonies. Importantly, the sham treatment led to immune alterations independent of, and sometimes opposite of burn-induced effects. PMID- 14752290 TI - Hydrogen peroxide derived from intestine through the mesenteric lymph induces lung edema after surgical stress. AB - Compelling evidence indicates that the small intestine is the primary source of factors inducing lung injury after major surgery and that the lymphatic system is the major route by which these gut-derived factors reach the pulmonary circulation. This study investigated the mechanism of lung edema induced by surgical stress. After subjecting male, fasted, pathogen-free Sprague-Dawley rats to surgical stress (laparotomy and intestinal handling for 5 min), followed by ventilation for 5 h, we measured H2O2 production in the mucosa of small intestine and in the lung using 2',7'-dichlorofluorescein and intravital fluorescence microscopy. In addition, H2O2 in mesenteric lymph was measured using a quantitative assay; lung permeability was assessed as a function of extravasation of Evans blue dye; neutrophil accumulation was visualized by intravital fluorescence microscopy and assessed as a function of myeloperoxidase activity; and TNF-alpha levels were measured using a specific ELISA. The intensity of 2',7' dichlorofluorescein fluorescence in the mucosa of small intestine, H2O2 levels of mesenteric lymph, and lung permeability were all significantly higher in rats subjected to surgical stress than in control animals. Moreover, all of these effects were blocked by pretreatment with a specific xanthine oxidase inhibitor. Surgical stress did not increase neutrophil accumulation or TNF-alpha production in the lung. In conclusion, surgical stress induces xanthine oxidase-dependent H2O2 production in the small intestine. The H2O2 then enters the mesenteric lymph and travels to the lung, where it increases capillary permeability and thus induces edema. PMID- 14752291 TI - The attenuation of hepatic microcirculatory alterations by exogenous substitution of nitric oxide by s-nitroso-human albumin after hemorrhagic shock in the rat. AB - Hepatic microcirculatory disorders such as narrowing of sinusoids after hemorrhagic shock play a major role in the pathogenesis of organ failure. It is known that the balance of vasoactive mediators such as endothelin and nitric oxide (NO) regulate microvascular perfusion, including the diameter of hepatic sinusoids. The present study was designed to evaluate the role of exogenous substitution of NO by S-nitroso-albumin (S-NO-HSA) in the prevention of pathophysiological alterations of hepatic microcirculation. Anesthetized Sprague Dawley rats were instrumented for invasive hemodynamic monitoring. Hemorrhagic shock was induced by bleeding to a mean arterial pressure (MAP) of 40 mmHg and was maintained for 60 min. Thereafter, the animals were resuscitated with shed blood and Ringer's solution. During the first hour of resuscitation, S-NO-HSA or pure HSA was infused continuously (10 micromol/kg/h) and hepatic microcirculation was detected by intravital epifluorescence microscopy either 5 or 24 h after the insult. Results were compared with a sham-treated group (n = 6-8 per group). Shock-induced microcirculatory narrowing of sinusoids was significantly reduced in the S-NO-HSA group compared with the HSA group both at 5 and 24 h (HSA: 9.3 +/ 0.2 microm; S-NO-HSA: 12.1 +/- 0.2 microm, P < 0.05). Sinusoidal perfusion was significantly higher in the S-NO-HSA group than in the HSA group (HSA: 50,934 +/- 1,382 microm3/s; S-NO-HSA: 78,120 +/- 2,348 microm3/s, P < 0.05). Reversible leukocyte adhesion to sinusoidal endothelium, an indicator of the inflammatory response, was significantly reduced in the S-NO-HAS-treated group. The findings of this study in a rat model of hemorrhagic shock suggest that NO substitution by S-NO-HSA during resuscitation attenuates both early and late hepatic microcirculatory disturbances as well as the increase in leukocyte adherence. PMID- 14752292 TI - P38 MAPK mediates myocardial proinflammatory cytokine production and endotoxin induced contractile suppression. AB - Cardiac myocytes are capable of synthesizing tumor necrosis factor alpha (TNF alpha), interleukin-1, and interleukin-6 (IL-1 and IL-6). p38 mitogen-activated protein kinase (MAPK) has been implicated in oxidant-stress-induced myocardial TNF-alpha production; however, the extent to which this kinase contributes to endotoxin-induced contractile dysfunction, as well as TNF-alpha, IL-1alpha, IL 1beta, and IL-6 production, in a bloodless model of endotoxin-induced myocardial dysfunction is unknown. Isolated rat hearts were perfused (Langendorff), and myocardial contractile function continuously recorded, during direct antegrade endotoxin infusion, with and without prior p38 MAPK inhibition. Ventricular p38 MAPK activation (phospho-p38 MAPK Western), cytokine mRNA (RT-PCR), and protein (ELISA) were determined. Endotoxin resulted in progressive decline in left ventricular developed pressure and coronary flow that was attenuated with prior p38 MAPK inhibition (SB 203580). p38 MAPK inhibition significantly decreased endotoxin-induced cardiac TNF-alpha, IL-1alpha, IL-1beta, and IL-6 mRNA levels. To determine the relative effect of TNF-alpha in inducing IL-1alpha, IL-1beta, and IL-6 production, TNF-alpha was sequestered during endotoxin infusion, and TNF alpha, IL-1beta, and IL-6 protein levels were measured. Interestingly, TNF-alpha sequestration alone significantly decreased myocardial IL-1beta and IL-6 production. We conclude that p38 MAPK is involved in endotoxin-induced myocardial contractile dysfunction and myocardial TNF-alpha production; however, p38 MAPK's involvement in IL-1 and IL-6 production may be indirectly mediated by TNF-alpha. PMID- 14752293 TI - Impairment of endothelium-dependent dilation response after resuscitation from hemorrhagic shock involved postreceptor mechanisms. AB - Resuscitation from hemorrhagic shock is associated with impairment of the endothelium-dependent dilation response, whereas the dilation response induced by the endothelium-independent pathway, which is mediated by nitroprusside, a nitric oxide (NO) donor and a direct activator of guanylate cyclase, remains unaltered. Whether the impairment of the endothelium-dependent dilation response is caused by a specific receptor alteration or generally a defect in signal transduction pathway remains undetermined. Anesthetized rats were monitored for hemodynamics, and the terminal ileum was prepared for intravital videomicroscopy. Hemorrhage was 50% of mean arterial pressure for 60 min followed by resuscitation with the shed blood returned plus 2 volumes of normal saline. Intestinal microvascular reactivity to the endothelium-dependent receptor-dependent agonists acetylcholine or substance P (10(-8) or 10(-6) M), as well as the endothelium-dependent receptor-independent calcium ionophore, was determined at baseline and at 2 h postresuscitation from hemorrhagic shock. Measured vascular diameters for premucosal A3 arterioles (pA3 and dA3) were normalized and expressed as percentage of the maximal dilation capacity, as obtained from the response to the endothelium-independent NO donor sodium nitroprusside (10(-4) M). At 2 h postresuscitation, there was a marked constriction of pA3 (-70.1 +/- 20) and dA3 (-61.5 +/- 11.6) from maximal dilation capacity. Baseline premucosal arteriolar response to substance P (10(-8) M) was 30.68 +/- 4.19% and 34.66 +/- 5.82% for pA3 and dA3 arterioles, respectively. This was significantly reduced to 20.97 +/- 2.41% and 17.94 +/- 3.60% at 2 h postresuscitation. However, no significant difference between baseline and postresuscitation arteriolar responses was observed at the higher dose of substance P (10(-6) M). Postresuscitation premucosal arteriolar response to the endothelium-dependent receptor-independent calcium ionophore (10(-9) to 10(-5) M) is characterized by a marked decrease in sensitivity and an enhanced threshold for calcium ionophore-mediated dilation. The logEC50 was -7.62 +/- 0.39 and -7.75 +/- 0.32 for the pA3 and dA3 at baseline, respectively. This was significantly (P < 0.01) reduced to -5.15 +/- 0.14 and -4.39 +/- 0.71 at 2 h postresuscitation. These data suggest that impairment of the endothelium-dependent dilation response after resuscitation from hemorrhagic shock is not mediated by specific receptor alteration. Cellular mechanisms that participate in or are part of oxygen free radical formation after resuscitation from hemorrhagic shock such as Ca2+ and leukocytes, appear to have a pivotal role in the mechanism of cellular dysfunction. PMID- 14752294 TI - Modulation of macrophage responsiveness to lipopolysaccharide by IRAK-1 manipulation. AB - Local activation of the macrophage by endotoxin is essential for the eradication of invasive gram-negative infections. Circulating endotoxin at lower concentrations results in immune cell activation at distant sites leading to tissue injury. Although the cellular mechanisms involved in these potentially dissimilar events are incomplete, it appears that the proximal kinase IRAK-1 plays a role. Thus, sense and antisense IRAK-1 oligonucleotides were used to determine the role IRAK-1 plays in macrophage activation by systemic (1-100 ng/mL) and local (1000 ng/mL) concentration of lipopolysaccharide (LPS) within THP-1 cells. Within the sense group, 1-1000 ng/mL of LPS within the sense group resulted in cellular activation of ERK-1/2, p38, and JNK/SAPK and the nuclear activation of NF-kappaB and AP-1. This activation was associated with proinflammatory cytokine production and cellular spreading. Systemic concentrations of LPS within the antisense group were associated with significant attenuation of intracellular signaling, cytokine production, and cellular spreading compared with the sense group. Local concentrations of LPS within the antisense group, however, were associated only with a delay in intracellular signaling, with no effect on cytokine production or cell spreading compared with the sense group. Based on these results, it appears that IRAK-1 is essential to macrophage activation at systemic, but not local, concentrations of LPS. These data suggest that redundant pathways exist that are functional at higher concentrations of LPS. Therefore, IRAK-1 appears to be the central kinase involved in the activation of the macrophage at distant sites during septic shock but is not necessary for activation in areas of local infection. PMID- 14752295 TI - Clinical trial design and outcomes in patients with severe sepsis. Steven M. Opal. Shock 20(4):295-302, 2003. PMID- 14752296 TI - Cardiopulmonary, histological and inflammatory alterations after lung contusion in a novel mouse model of blunt chest trauma. Shock 19(6):519-525, 2003. PMID- 14752297 TI - Memorable medical mentors: I. Lewis Hill Weed (1886-1952). PMID- 14752298 TI - The trials and tribulations of long-term postmenopausal hormone therapy. PMID- 14752299 TI - Subfertility and gastrointestinal disease: 'unexplained' is often undiagnosed. AB - Subfertility can be more reliably explained and effectively treated with an improved understanding of the contribution of chronic medical disease to reproductive dysfunction. This review addresses several common gastrointestinal disorders which are increasingly implicated in infertility and early pregnancy loss: celiac disease, inflammatory bowel disease (ulcerative colitis and Crohn's disease), and hemochromatosis. Appreciating the reproductive impact of these comorbidities and their treatments enables clinicians to accurately counsel patients and to modify medical and fertility treatments based on etiology. Because unexplained infertility can represent the initial presentation of undiagnosed medical disease, considering these often-subclinical gastrointestinal disorders in the differential diagnosis of subfertility provides an opportunity not only to increase the probability of conception and uncomplicated pregnancy, but also to improve overall maternal health. PMID- 14752300 TI - Metformin in obstetric and gynecologic practice: a review. AB - Metformin is a common treatment for women who have insulin resistance manifesting as type 2 diabetes or polycystic ovarian syndrome (PCOS). With an increasing number of these patients conceiving, it is expected that the use of metformin in and around the time of pregnancy will increase. This article reassesses the mechanisms, safety, and clinical experience of metformin use in obstetrics and gynecology. Metformin is an attractive therapeutic option because administration is simple, hypoglycemia rare, and weight loss promoted. There is a large volume of research supporting the use of metformin treatment in diabetes mellitus, androgenization, anovulation, infertility, and recurrent miscarriage. Although metformin is known to cross the placenta, there is, as yet, no evidence of teratogenicity. Metformin has an array of complex actions, accounting for the varied clinical roles, many of which are still to be fully evaluated. Much research is still needed. PMID- 14752301 TI - Misoprostol: a quarter century of use, abuse, and creative misuse. AB - Misoprostol is a prostaglandin E1 analog originally intended for use to prevent NSAID-induced gastric ulcers. However, because of its cervical ripening and uterotonic property, misoprostol has become one of the most useful drugs in obstetrics and gynecology. Misoprostol has proven to be a very convenient and flexible drug because of its formulation as a tablet that is stable and that can be administered orally, rectally, vaginally and by the sublingual route. Beginning with its abuse for illegal abortion in the late 1980s, misoprostol has quickly become established as one of the most effective drugs for terminating pregnancies in the first and second trimesters, as well as for inducing labor in the third trimester. Its use for routine prevention of postpartum hemorrhage has not been so successful, partly as the high doses required for this indication often result in troublesome side effects. Despite the large body of medical evidence about its efficacy and relative safety, the use of misoprostol in pregnant women remained off-label until the spring of 2002. PMID- 14752302 TI - Detecting insulin resistance in polycystic ovary syndrome: purposes and pitfalls. AB - Approximately 50% to 70% of all women with polycystic ovary syndrome (PCOS) have some degree of insulin resistance, and this hormone insensitivity probably contributes to the hyperandrogenism that is responsible for the signs and symptoms of PCOS. Although uncertainty exists, early detection and treatment of insulin resistance in this population could ultimately reduce the incidence or severity of diabetes mellitus, dyslipidemia, hypertension, and cardiovascular disease. Even if that proves to be the case, there are still several problems with our current approach to insulin sensitivity assessment in PCOS, including the apparent lack of consensus on what defines PCOS and "normal" insulin sensitivity, ethnic and genetic variability, the presence of other factors contributing to insulin resistance such as obesity, stress, and aging, and concern about whether simplified models of insulin sensitivity have the precision to predict treatment needs, responses, and future morbidity. Although the hyperinsulinemic-euglycemic clamp technique is the gold standard for measuring insulin sensitivity, it is too expensive, time-consuming, and labor-intensive to be of practical use in an office setting. Homeostatic measurements (fasting glucose/insulin ratio or homeostatic model assessment [HOMA] value) and minimal model tests (particularly the oral glucose tolerance test [OGTT]) represent the easiest office-based assessments of insulin resistance in the PCOS patient. The OGTT is probably the best simple, office-based method to assess women with PCOS because it provides information about both insulin resistance and glucose intolerance. The diagnosis of glucose intolerance holds greater prognostic and treatment implications. All obese women with PCOS should be screened for the presence of insulin resistance by looking for other stigmata of the insulin resistance syndrome such as hypertension, dyslipidemia, central obesity, and glucose intolerance. PMID- 14752303 TI - Perspective: rejuvenation of the periocular region: a unified approach to the eyebrow, midface, and eyelid complex. PMID- 14752304 TI - Review: primary epithelial malignancies of the lacrimal gland: the 2003 Ramon L. Font lecture. PMID- 14752305 TI - Outcomes in patients with adenoid cystic carcinoma of the lacrimal gland. AB - PURPOSE: To evaluate the outcomes among patients with adenoid cystic carcinoma of the lacrimal gland treated at various stages of their disease at a tertiary care cancer center. METHODS: A retrospective case series of 20 patients with adenoid cystic carcinoma of the lacrimal gland treated at a single institution between 1952 and 2002. Clinical records were available for all 20 patients; histologic sections from 12 of the 20 patients were available for review. Disease-free survival was measured from the completion of treatment; overall survival was measured from the date of initial diagnosis. RESULTS: The study included 6 men and 14 women. The mean age at diagnosis was 39.5 years. The median follow-up time was 34 months (range, 6 to 264 months). The local/regional treatment modalities included exenteration with bone removal and radiation therapy (RT) in 5 patients, exenteration with RT (no bone removal) in 8 patients, exenteration (no RT or bone removal) in 1 patient, exenteration with bone removal (no RT) in 1 patient, local resection with RT in 3 patients, and local resection without RT in 2 patients. Overall, 16 patients had RT as part of their treatment regimen. Seven patients (35%) had local recurrence. Sixteen patients (80%) had distant metastasis during the study period. At the time of this report, 13 (65%) of the patients had died of disease. The median disease-free survival for the entire group was 18 months. Eight patients had a predominantly basaloid histologic pattern. Ten patients had verifiable histologic evidence of perineural invasion. CONCLUSIONS: This study underscores the generally grave prognosis for patients with adenoid cystic carcinoma of the lacrimal gland and the difficulty in making any conclusive recommendations for local therapy for this disease. PMID- 14752306 TI - Intraosseous hemangioma of the lateral orbital wall. AB - PURPOSE: To report the clinical and radiologic features of two patients with intraosseous orbital hemangiomas of the lateral wall. METHODS: Case reports and literature review. RESULTS: The first patient presented with an indolent mass over the lateral orbital rim. Intraosseous hemangioma was not considered in the differential diagnosis. A biopsy specimen was taken from the tumor, which caused significant bleeding. After biopsy, an orbital CT scan revealed a lesion involving the left lateral orbital rim and soft tissue of the temporal fossa. The rim was grossly enlarged, with several cavities of different sizes. The tumor of the second patient was located more posteriorly and extended into the roof. The patient complained of parietal headache, epiphora, and local pain. A CT scan revealed a round, lytic lesion involving the greater sphenoid wing and frontal bone. The tumor was approached through a coronal flap and resected en bloc without problems. CONCLUSIONS: The diagnosis of intraosseous hemangioma can be difficult, especially when tumors are located in the anterior part of the lateral wall. In this location, intraosseous hemangiomas tend to induce few symptoms and thus have a higher likelihood of biopsy being performed before a correct diagnosis. A CT scan with bone windows can play a beneficial role in the evaluation of lytic bone lesions in intraosseous hemangioma. PMID- 14752307 TI - Sebaceous gland tumors of the eyelids and conjunctiva in the Muir-Torre syndrome: a clinicopathologic study of five cases and literature review. AB - PURPOSE: To study the sebaceous tumors of eyelid/conjunctiva associated with Muir Torre syndrome (MTS) and to determine the role of immunohistochemical markers (MSH2, mismatch repair gene) in the diagnosis of patients with MTS. METHODS: A retrospective, noncomparative case series of 5 patients diagnosed with MTS from our laboratory. We also reviewed all previously reported cases of sebaceous eyelid tumors with a visceral malignancy. RESULTS: Four of the 5 patients were men, with a mean age of 55 years (range, 41 to 76 years). Four of the 5 patients had gastrointestinal carcinoma. On histopathological examination, 4 of the 5 tumors were classified as sebaceous adenomas that exhibited a distinct lobular pattern with prominent basaloid cells at the periphery of the lobules. One tumor was classified as a well-differentiated sebaceous gland adenocarcinoma. The diagnosis of MTS in all 5 patients was made after the diagnosis of the eyelid lesions. Immunohistochemical stains showed a lack of MSH2 expression in two tumors, which is highly consistent with MTS. CONCLUSIONS: Muir-Torre syndrome should be considered in patients who develop sebaceous tumors of the ocular adnexa. Immunohistochemistry for MSH2 is a practical initial approach for screening MTS in patients with sebaceous tumors. PMID- 14752308 TI - Tarsal strip procedure for the correction of tearing. AB - PURPOSE: To evaluate the tarsal strip procedure in the treatment of tearing related to lacrimal pump failure. METHODS: A retrospective chart review of all tearing patients undergoing the tarsal strip procedure was done. Patients with ectropion, nasolacrimal duct obstruction, and punctal eversion and stenosis were excluded. Data collected included patient age, sex, procedures performed, follow up, and resolution of tearing. The main outcome measure is resolution of tearing. RESULTS: Thirty-four eyelids of 21 patients underwent a tarsal strip procedure for the correction of tearing. Complete resolution of tearing was noted in 14 eyes. Seventeen eyes were partially improved and required no further intervention. Three eyes were unimproved over the follow-up period. Two eyes eventually required another procedure. CONCLUSIONS: Patients undergoing tarsal strip for tearing caused by presumed lacrimal pump failure showed a significant rate of improvement in their tearing symptom after the procedure. The tarsal strip procedure appears to be effective in the surgical treatment of tearing secondary to laxity of the lower eyelid. PMID- 14752309 TI - A new, wider-diameter Crawford tube for stenting in the lacrimal drainage system. AB - PURPOSE: To determine whether technology is available to secure a larger-diameter Crawford tube swedged onto the metal Crawford introducer and to use the system in a closed intubation to treat a failed dacryocystorhinostomy. METHODS: A metal ferrule has been developed to swedge a larger-diameter Crawford tube onto the standard Crawford probe. Seven patients with failed dacryocystorhinostomy procedures were treated with probing and intubation with the use of this larger tube Crawford system. RESULTS: Six of the 7 patients had patent lacrimal systems after the tubes were removed, a result comparable to open revision after a failed dacryocystorhinostomy. In each case, on insertion, the tube remained firmly attached to the Crawford probe and did not dislodge. CONCLUSIONS: The technology of using a metal ferrule at the junction of the standard Crawford probe and a larger silicone tube was successful in creating a Crawford system with greater dilation capability than the standard Crawford system. The technique of probing and intubation with this larger Crawford system may be a simple alternative to open revision of a failed dacryocystorhinostomy procedure. PMID- 14752310 TI - Double silicone intubation as treatment for persistent congenital nasolacrimal duct obstruction. AB - PURPOSE: The great majority of children with nasolacrimal duct (NLD) obstruction are successfully treated with probing or conventional silicone tube intubation. A small percentage of patients fail to have their NLD obstruction resolved with these procedures and require dacryocystorhinostomy (DCR). This study was conducted to assess the effectiveness of double bicanalicular silicone intubation with placement of two loops of silicone tubing through the NLD for treatment of persistent NLD obstruction in children as an alternative to DCR. METHODS: Ten eyes of 9 patients with nasolacrimal duct obstruction who had failed conventional therapies and were to otherwise undergo DCR were instead treated with double bicanalicular silicone tube intubation. Resolution of preoperative symptoms and signs of NLD obstruction were assessed. RESULTS: At an average follow-up of 40 +/ 5.6 months, all patients had improvement in symptoms and signs. The mean patient age was 31 +/- 3.2 months. The average duration of intubation was 15 +/- 0.73 months. Eight of 10 eyes had complete resolution of NLD obstruction symptoms. Two of 10 treated eyes had occasional symptoms of mattering with upper respiratory infection and exposure to wind or cold but required no further treatment. Nine of 10 eyes were treated for congenital NLD obstruction. The remaining patient had failed conventional bicanalicular intubation after repair of a traumatic common canalicular avulsion. CONCLUSIONS: Double bicanalicular silicone tube intubation is an effective alternative to DCR in selected children who have failed conventional therapies for NLD obstruction. This treatment obviated DCR in all patients in this study. PMID- 14752311 TI - Mechanical endonasal dacryocystorhinostomy versus external dacryocystorhinostomy. AB - PURPOSE: To compare the success rates of a new mechanical endonasal dacryocystorhinostomy (MENDCR) technique and the conventional external dacryocystorhinostomy technique (Ext-DCR). METHODS: A prospective, nonrandomized interventional comparative case series of 31 consecutive MENDCRs and 24 Ext-DCRs performed from January 1999 to December 2000. Patients with anatomic nasolacrimal duct obstruction were included in the study; previous lacrimal surgery and functional nasolacrimal and canalicular obstruction were exclusion criteria. Two surgeons performed the MENDCRs, using a standardized operative technique, which involved creation of a large bony ostium and mucosal flaps between the lacrimal sac mucosa and nasal mucosa. One surgeon performed all Ext-DCRs. RESULTS: Twenty seven patients (8 men, 19 women) underwent 31 MENDCRs. The average age of the patients was 62.9 years (range, 15 to 86 years; SD, 19.1 years). In 11 cases (35.4%), a septoplasty was required at the time of surgery, and in 6 cases (19%), further endoscopic sinus surgery was performed. In the Ext-DCR group, 23 patients (7 men, 16 women) underwent 24 DCRs. The average age was 59.6 years (range, 22 to 86 years; SD, 18.5 years). No other nasal procedures were performed at the time of surgery in this group. The average follow-up time was 13 months for the MENDCR group and 12.4 months for the Ext-DCR group. Success was defined as relief of symptoms and by anatomic patency, which was assessed by history, fluorescein flow on nasal endoscopy, and lacrimal syringing. In the MENDCR group, surgery was successful in 29 of 31 DCRs (93.5%); 1 of 2 failed cases was anatomically patent but symptomatic, yielding an anatomic patency rate of 96.8%. In the Ext-DCR group, the success rate was 95.8% (23/24 DCRs); the failed case was anatomically patent but symptomatic, giving an anatomic patency rate of 100%. The differences in overall success and anatomic patency were not statistically significant (P = 0.6 and P = 0.56, 1-tailed Fisher exact test). CONCLUSIONS: The success rate of MENDCR (93.5%) compares favorably with that of standard external DCR (95.8% in this study). MENDCR relies on the creation of a large ostium and mucosal flap apposition. A larger, randomized prospective trial is needed to fully assess the efficacy of this new technique. PMID- 14752312 TI - External dacryocystorhinostomy and local anesthesia: technique to measure minimized blood loss. AB - PURPOSE: To describe the techniques required to minimize blood loss in external dacryocystorhinostomy (DCR) under local anesthesia and to accurately measure the intraoperative blood loss through application of these techniques. METHODS: Prospective single-surgeon series to include the first 20 sequential external DCRs from June 2002. Hemostasis was maximized at each stage of the procedure. Blood loss was measured by attaching a collection pot to the first link in the vacuum tubing. Patient demographics and operative duration were recorded. RESULTS: The mean blood loss was 4.5 mL (range, 1 to 14 mL). The mean operative duration was 36 minutes (range, 25 to 65 minutes). Seventy-five percent of patients were female and 25% were male. The mean age was 57 years (range, 48 to 76 years). CONCLUSIONS: Intraoperative blood loss in external DCR can be reduced to a negligible level through careful patient preparation and operative technique. Minimal blood loss allows for patient comfort with DCR under local anesthesia. PMID- 14752313 TI - Orbital trauma caused by bicycle hand brakes. AB - PURPOSE: This report aims to increase awareness of an unusual mechanism of orbital injury sustained by bicycle riders. METHODS: In this retrospective small case series, we describe two cases of orbital injury caused by upper eyelid penetration. A 5-year-old boy (patient 1) and a 6-year-old boy (patient 2) presented to our service within a 2-week period. Both had been injured by similarly styled, handlebar-mounted bicycle hand brake levers. Patient 1 had an orbital roof fracture and penetrating brain injury and underwent repair of a left upper eyelid laceration, craniotomy for pseudoencephalocele, and ptosis repair. Patient 2 had orbital hemorrhage and underwent repair of left upper eyelid laceration. RESULTS: In both cases, a handlebar-mounted bicycle hand brake lever perforated the left eyelid when the rider fell onto it. Neither patient was wearing protective headwear or eyewear. Two months after surgery, patient 1 had 20/25 visual acuity OU and excellent cosmetic appearance. Patient 2 had baseline amblyopic vision 2 days after surgery but moved from town and was lost to follow up. CONCLUSIONS: Orbit injuries from bicycle brake levers are rare, and helmets or protective eyewear probably would not have prevented these injuries. However, a change in the design and/or mounting location of handlebar-mounted brake levers might help prevent further injuries of this type. PMID- 14752314 TI - Consecutive ectropion after the Wies procedure. AB - PURPOSE: To describe the clinical features and management difficulties of the ectropions after Wies procedure for lower eyelid involutional entropion in 4 patients. METHODS: Case series. RESULTS: All ectropions had developed within 2 months of the Wies procedure. Common structural features with cicatricial contraction of the anterior lamella and buckling of the tarsal plates were evident. Maximal horizontal eyelid shortening and excision of anterior lamella scar tissue allowed correction. CONCLUSIONS: Consecutive ectropion is caused by secondary cicatricial changes, which can be difficult to manage. Untreated horizontal laxity and postoperative scarring are likely contributory factors in its development. The authors recommend excision of anterior lamella scar tissue combined with large full-thickness wedge excision as an effective treatment. PMID- 14752315 TI - Coralline hydroxyapatite orbital implant (bio-eye): experience with 158 patients. AB - PURPOSE: To assess the problems seen in 158 patients with coralline hydroxyapatite (HA) orbital implants (Bio-Eye). METHODS: A consecutive case series of 170 patients receiving coralline HA implanted by two surgeons over a 5 year period were reviewed. The authors analyzed age, type of surgery, implant size, peg system, follow-up duration, time of pegging, problems encountered, and treatment. RESULTS: Twelve patients were lost to follow-up after 5 months, leaving 158 patients who were followed from 6 to 130 months (average, 39 months). Problems in unpegged implants occurred in 36 (22.8%) patients. Discharge occurred in 18 (11.4%) patients, implant exposure in 12 (7.6%), socket discomfort in 1 (0.6%), conjunctival thinning in 3 (1.9%), chronic conjunctival swelling in 2 (1.3%), and implant infection in 3 (1.9%). Problems after pegging occurred in 68 (50.7%) of 134 patients: discharge in 27 (20.1%), pyogenic granuloma in 24 (17.9%), conjunctiva overgrowing the peg in 4 (3.0%), implant exposure around the sleeve in 5 (3.7%), clicking in 6 (4.5%), peg on an angle in 2 (1.5%), loose sleeve in 1 (0.7%), peg falling out in 18 (13.4%), popping peg in 1 (0.7%), poor transfer of movement in 3 (2.2%), pain with movement in 1 (0.7%), and implant infection in 2 (1.5%). CONCLUSIONS: The Bio-Eye orbital implant represents a porous orbital implant that is biocompatible with orbital tissues and allows fibrovascular ingrowth and improved motility when coupled to the overlying artificial eye. It is more expensive than other commercially available porous orbital implants, such as synthetic FCI3 HA, porous polyethylene (Medpor), and aluminum oxide (Bioceramic) implant. Problems encountered with its use are similar to those problems seen in patients with the synthetic FCI3 hydroxyapatite and aluminum oxide orbital implants. PMID- 14752316 TI - Evolution of an orbital apex tumor over a decade. AB - We report the evolution of an orbital apex tumor that underwent spontaneous regression, followed several years later by rapid growth and loss of vision. A turkey farmer had a right apical lesion, initially thought to be an optic nerve meningioma. The mass spontaneously regressed over 2 years, with marked improvement of vision. Eight years later, the patient presented with loss of light perception and severe proptosis of the right eye from a massive orbital tumor that extended in the cavernous sinus. An eyelid-sparing orbital exenteration was performed. The pathologic diagnosis was diffuse large cell lymphoma. This case demonstrates that non-Hodgkin lymphoma of the orbital apex may resolve spontaneously or it may enlarge rapidly, degenerating into more aggressive forms. PMID- 14752318 TI - Phantom vision: a graphic description. AB - An 80-year-old man, who is an experienced artist, underwent orbital exenteration for invasive squamous cell carcinoma. After exenteration, the patient noted a persistent, colored visual phenomenon. The phantom vision continued unchanged to 9 months after surgery (the time of writing), when the patient presented a graphic and written description of phantom vision. PMID- 14752317 TI - Primary pelvic telangiectatic osteosarcoma metastatic to both orbits. AB - We report a case of a 23-year-old man with primary pelvic telangiectatic osteosarcoma metastatic to both orbits. The patient had proptosis with optic neuropathies and intermittent third nerve palsies. His disease was unresponsive to chemotherapy or radiotherapy, and extensive craniofacial involvement precluded surgical resection. The patient died of his disease. Telangiectatic osteosarcoma, a rare variant of osteosarcoma that is distinguished by blood-filled cystic spaces, may metastasize to the orbit and skull base. Because telangiectatic osteosarcoma may radiographically resemble other benign and malignant lesions, biopsy is essential for accurate diagnosis that will ultimately dictate clinical therapy. However, patients with this disease have a poor prognosis. PMID- 14752319 TI - Intrasinus wood foreign body causing orbital cellulitis in congenital insensitivity to pain with anhidrosis syndrome. AB - A 2-year-old boy with congenital insensitivity to pain with anhidrosis (CIPA) was referred with a 2-day history of left periorbital swelling and mucoid conjunctival discharge. Marked worsening was noted despite intramuscular ceftriaxone treatment for 3 days, with marked proptosis, conjunctival chemosis, and a frozen eye. Orbital cellulitis was suspected. Ceftriaxone was intravenously administered. Orbital computed tomography (CT) disclosed an inflammatory process in the medial aspect of the left orbit with ethmoiditis. Improvement was not noted after external ethmoidectomy and drainage of the intraconal abscess. Repeat CT showed a recurrent intraconal abscess. A revision external ethmoidectomy was performed, and a bent wooden match was removed from the posterior aspect of the ethmoidal sinus, after which significant clinical improvement was noted. In patients with CIPA, accidental or self-penetrated foreign bodies must be kept in mind when treating all types of wounds. PMID- 14752320 TI - Asymptomatic lacrimal ductule dacryolithiasis with embedded cilia. AB - We describe a case of a 32-year-old man who presented with a visually apparent but otherwise asymptomatic mass in the right lateral fornix. Computerized tomography demonstrated the mass adjacent to the lacrimal gland. The mass was surgically removed, and histopathologic examination was consistent with a dacryolith of the lacrimal ductule. There were several cilia isolated from the dacryolith. This entity should be considered in the differential diagnosis of patients presenting with a localized mass in the region of the lacrimal gland. PMID- 14752321 TI - Complications of bioabsorbable orbital implants and fixation plates. AB - After orbital floor and trimalar fracture repair with the use of a bioabsorbable maxillofacial implant, a patient had eyelid retraction with a thickened, immobile eyelid. We observed a dense, fibrous scar and encapsulation at the implantation site. PMID- 14752322 TI - Subperiosteal hematoma after surgical treatment for subarachnoid hemorrhage. AB - After the surgical treatment for subarachnoid hemorrhage, computed tomography and magnetic resonance imaging clearly demonstrated a fracture in the medial wall of the left orbit and a superonasal subperiosteal hematoma. Removal of the subperiosteal hematoma and correction of the fracture were successful. Subperiosteal hematoma is an unusual complication after inadvertent neurosurgical manipulation. PMID- 14752325 TI - The right mix: using nutritional interventions and an anabolic agent to manage a stage IV ulcer. PMID- 14752324 TI - Growth factors and chronic wound healing: past, present, and future. AB - Growth substances (cytokines and growth factors) are soluble signaling proteins affecting the process of normal wound healing. Cytokines govern the inflammatory phase that clears cellular and extracellular matrix debris. Wound repair is controlled by growth factors (platelet-derived growth factor [PDGF], keratinocyte growth factor, and transforming growth factor beta). Endogenous growth factors communicate across the dermal-epidermal interface. PDGF is important for most phases of wound healing. Becaplermin (PDGF-BB), the only growth factor approved by the Food and Drug Administration, requires daily application for neuropathic wound healing. Gene therapy is under development for more efficient growth factor delivery; a single application will induce constitutive growth factor expression for weeks. Based on dramatic preclinical animal studies, a phase 1 clinical trial planned on a PDGF genetic construct appears promising. PMID- 14752323 TI - Wound pain: a challenge for the patient and the wound care specialist. AB - PURPOSE: To provide physicians and nurses with an overview of the mechanisms, pathophysiology, assessment, and treatment of pain related to pressure ulcers. TARGET AUDIENCE: This continuing education activity is intended for physicians and nurses with an interest in learning about management of patients with pressure ulcer-related pain. LEARNING OBJECTIVES: After reading the article and taking the test, the participant will be able to: 1. Describe the mechanisms and pathophysiology of pain related to pressure ulcers. 2. Identify assessment parameters and treatment options for pain related to pressure ulcers. PMID- 14752326 TI - 2004 Medicare changes impact hospital-owned outpatient wound departments. PMID- 14752327 TI - How to make dressing changes less painful. PMID- 14752328 TI - Steps for determining the toe brachial pressure index. PMID- 14752329 TI - V(D)J recombination and DNA repair: lessons from human immune deficiencies and other animal models. AB - PURPOSE OF REVIEW: V(D)J recombination not only represents the main mechanism for the diversification of the immune system, it also constitutes a critical checkpoint in the development of both B and T lymphocytes. While a defect in V(D)J recombination leads to severe combined immune deficiency, a deregulation of this process can participate in the onset of lymphoid malignancies. RECENT FINDINGS: The careful analysis of human severe combined immune deficiency patients as well as engineered murine models provided several new interesting insights into the physiopathology of the V(D)J recombination process. A new factor of the V(D)J recombination/DNA repair machinery, Artemis, was identified based on its deficiency in human severe combined immune deficiency patients. It also became evident from knockout mouse studies that DNA repair factors that participate in V(D)J recombination can be considered as genomic caretakers. SUMMARY: While V(D)J recombination was first recognized as a critical checkpoint in the development of the immune system, the discovery of several DNA repair factors that participate in this reaction shed light on more general aspects of genomic stability and cancer predisposition. PMID- 14752330 TI - Immune dysregulation, polyendocrinopathy, enteropathy, X-linked syndrome: a model of immune dysregulation. AB - PURPOSE OF REVIEW: Immune dysregulation, polyendocrinopathy, enteropathy, X linked syndrome is a fatal syndrome of overwhelming autoimmunity. Recent identification of FOXP3 as the causative gene and realization that this same gene defect occurs in the mutant mouse Scurfy has yielded new insights and hopes of unraveling the mechanism of autoimmunity in this and possibly other diseases. In this review, we describe the clinical features of immune dysregulation, polyendocrinopathy, enteropathy, X-linked syndrome/Scurfy and compare this phenotype with similar syndromes caused by other single gene mutations. We examine therapeutic options to treat the syndrome, study its immunologic basis, and investigate the structure and function of the FOXP3 protein. RECENT FINDINGS: The Scurfy mutant mouse has a characteristic phenotype that causes death by approximately 3 weeks of age. It is known that the effector cells in the Scurfy mouse are CD4+ T cells and that a population of normal T cells can control the overwhelming autoimmunity that they induce. Recent data have demonstrated that this process requires antigenic stimulation and that the degree to which the immune system responds is inversely proportional to the level of FOXP3 protein (Forkhead box P3) expression in peripheral T cells. Suppression of immune activation by FOXP3 may occur due to its ability to bind to DNA through a putative forkhead DNA-binding motif and to repress transcriptional activation from certain promoters in T cells. SUMMARY: Because of the dramatic phenotype and rapidity of onset, immune dysregulation, polyendocrinopathy, enteropathy, X linked syndrome and Scurfy provide a powerful model in which to study mechanisms of T cell regulation. A more complete understanding of this syndrome will provide important insights into mechanisms of immune suppression, tolerance, and autoimmunity. PMID- 14752331 TI - Cross-talk between CD40 and CD40L: lessons from primary immune deficiencies. AB - PURPOSE OF REVIEW: The purpose of this review is to provide an update of the molecular bases of CD40-mediated signalling and of the human immune defects associated with abnormalities of this activation pathway. RECENT FINDINGS: Over the last years considerable progress in the identification of intracellular molecules mediating CD40 signalling has been achieved. This review focuses on the recent work on the molecular mechanisms of CD40 signalling mediated by tumor necrosis factor receptor-associated factors, by transcription of the activation induced cytidine deaminase gene and by activation of nuclear factor kappa B. Furthermore, the importance of CD40/CD40L interaction for the induction of adaptive immunity will be outlined in the context of primary immunodeficiencies due to defects of the genes involved in the CD40 signalling pathway, which are characterized by an immunological phenotype of hyper-IgM syndrome. SUMMARY: The critical role of CD40/CD40L interactions in the development of various disease states has been fully appreciated, and further understanding of the molecular events involved in CD40 signalling may allow the identifications of candidate genes for other hyper-IgM syndromes. Molecular diagnosis will help to provide the most appropriate treatment and prognosis. PMID- 14752332 TI - Cytokines and their role in lymphoid development, differentiation and homeostasis. AB - PURPOSE OF REVIEW: The development of lymphoid tissues as well as the ultimate differentiation of naive and memory T cells are dependent on cytokines. In this review, we will focus on recent advances in the understanding of molecular mechanisms that regulate lymphoid development, homeostasis and tolerance. RECENT FINDINGS: Cytokines play a critical role in the development and differentiation of lymphoid cells. In addition, newer data indicate important roles of interleukin-7 and interleukin-15 in lymphoid homeostasis and memory. Furthermore, a new family of heterodimeric cytokines comprising interleukin-12, interleukin-23 and -27 is important for differentiation of helper T cells and cell-mediated immunity. Finally the importance of tumor necrosis factor superfamily members in the development of lymphoid organs has recently been elucidated and will be discussed in detail. SUMMARY: New cytokines and receptors continue to be identified. The discovery and characterization of cytokines, their receptors and signaling molecules will provide a more complete understanding of normal lymphoid development, differentiation and function. In addition, this knowledge should improve our understanding of the pathogenesis of immunological diseases and hopefully will provide new treatment strategies. PMID- 14752333 TI - Gene therapy of X-linked severe combined immunodeficiency. AB - This review addresses several questions in the light of the results recently obtained by a gene therapy trial for the treatment of X-linked severe combined immunodeficiency. This primary immunodeficiency, characterized by a complete absence of T and natural killer lymphocytes, appeared as a good model for the application of gene therapy, combining an expected selective advantage for transduced cells, an absence of immunological response to the vector and/or the therapeutic transgene together with accessibility to hematopoietic stem cells. After a brief description of the disease and its physiopathology we summarize the clinical results of the gene therapy trial putting them in perspective with those obtained following allogeneic hematopoietic stem cell transplantation. Definitive conclusions cannot be thrown due to the limited number of gene therapy-treated patients and their relatively short follow-up. PMID- 14752334 TI - The autoinflammatory syndromes. AB - PURPOSE OF REVIEW: To review the remarkable recent progress in our understanding of a range of inflammatory conditions in humans that until recently appeared unrelated. The term autoinflammatory disease has been proposed to describe a group of disorders characterized by attacks of seemingly unprovoked inflammation without significant levels of autoantibodies and autoreactive T cells. RECENT FINDINGS: As the link between the innate immune response and disease susceptibility has become more apparent, some remarkable associations have emerged. The majority of hereditary periodic fevers are due to mutations in the pyrin and tumour necrosis factor receptor superfamilies of molecules, both of which are intimately involved in innate immunity. Pyrin/marenostrin protein is mutated in familial Mediterranean fever, while mutations in a related protein, cryopyrin, are associated with Muckle-Wells/familial cold urticaria and chronic infantile neurologic cutaneous and articular syndrome. Both of these proteins interact with the apoptotic speck-like protein involved in caspase-1 activation and regulation of nuclear factor kappa B transcription; furthermore cryopyrin contains regions of homology with the nucleotide-binding oligomerization domain 2 protein, which is associated with susceptibility to Crohn's disease. Variants in the leucine-rich repeat domain of nucleotide-binding oligomerization domain are found in approximately 20% of patients with Crohn's disease, depending on ethnic background, while mutations in the NACHT domain are associated with a rare dominant granulomatous disease called Blau syndrome. SUMMARY: The study of autoinflammatory disease has progressed from genetics to definition of the functional defects in these patients. Although a direct association between defective innate immune responses to bacterial components and these diseases has not been formally established, much ongoing research is aimed towards confirmation of that hypothesis. PMID- 14752335 TI - Early defects in B cell development. AB - PURPOSE OF REVIEW: Recent clinical studies in patients with genetically proven X linked or autosomal recessive agammaglobulinemia provide some guidelines that should influence our management of patients with suspected immunodeficiency. RECENT FINDINGS: Males who are at a high risk of having X-linked agammaglobulinemia because they have an affected brother or uncle are often not evaluated for immunodeficiency until they are hospitalized for infection. Some of those who are evaluated are not started on gammaglobulin therapy immediately. More than 10% of patients with X-linked agammaglobulinemia are hospitalized for infection at less than 6 months of age, indicating that patients with known X linked agammaglobulinemia should be started on therapy by 2-3 months of age. In patients with sporadic X-linked agammaglobulinemia, the incidence of chronic lung disease correlates with the age at diagnosis, highlighting the importance of early diagnosis. Although almost all patients who are diagnosed as having the condition at more than 12 months of age have a history of recurrent otitis, 93% are not evaluated for immunodeficiency until they are hospitalized for infection. Because the physical exam provides a clue to the diagnosis of the condition- unusually small or absent cervical lymph nodes and tonsils--it should be possible to make an early diagnosis in a greater percentage of patients. Patients with autosomal recessive agammaglobulinemia have an earlier onset of disease compared with patients with X-linked agammaglobulinemia and they are more likely to have severe complications of the disease. SUMMARY: There is plenty of room for improvement in the diagnosis and management of patients with defects in early B cell development. PMID- 14752336 TI - Is sublingual immunotherapy clinically effective? AB - PURPOSE OF REVIEW: Sublingual immunotherapy has become increasingly popular and in some countries more allergic patients are treated by the sublingual route than the subcutaneous route. Evaluation of the scientific documentation for clinical efficacy is important before a treatment is used without restriction. This review critically analyses every placebo-controlled, double-blind study providing symptom/medication scores for the primary outcome; that is, a significant and clinically relevant reduction in disease severity of actively treated patients. RECENT FINDINGS: In total 23 papers fulfill the evaluation criteria; 26% are categorized as unequivocally effective, 35% are possibly effective (significant improvement in either symptom or medication scores), and 39% have no statistically documented efficacy. SUMMARY: The majority of papers have used an inadequate study design that may be responsible for the large number of inconclusive and negative studies. Before sublingual immunotherapy can be recommended as a routine treatment, more documentation for beneficial efficacy is needed. PMID- 14752337 TI - Respirable antisense oligonucleotides: a new, third drug class targeting respiratory disease. AB - PURPOSE OF REVIEW: To describe the potential of a new class of respiratory drugs, respirable antisense oligonucleotides. RECENT FINDINGS: The first respirable antisense oligonucleotide, EPI-2010, has now reached clinical trials. It has shown intriguing initial indications of efficacy and the potential to be the first once-per-week asthma preventative. Respirable antisense oligonucleotides are capable of addressing targets that have proven to be intractable to traditional 'small molecule' approaches, and against which newer monoclonal antibody strategies may also not be optimal. Respirable antisense oligonucleotides functionally, but not genetically, ablate gene expression by blocking the template function of target respiratory messenger RNAs by as yet incompletely defined mechanisms. They do so with an avidity and specificity which can be several orders of magnitude greater than those shown by small molecule antagonists for their protein targets. The target properties of respiratory messenger RNAs are strikingly different from those of respiratory proteins, enabling respirable antisense oligonucleotides to offer the potential of longer duration of effect, increased specificity of effect, and lack of systemic side effects compared with either traditional small molecule protein antagonists or monoclonal antibodies. SUMMARY: Respirable antisense oligonucleotides represent a new, third class of respiratory drugs with the potential to extend the range of therapeutic responses to otherwise intractable respiratory targets, and to address precedented targets with the possibility of improving on such features as safety and durability of response. PMID- 14752338 TI - Is immunotherapy effective in allergic disease? AB - PURPOSE OF REVIEW: The purpose of this review is to summarize the basic principles of effective immunotherapy, and to highlight some of the advances that have been published in the past year. RECENT FINDINGS: Recent findings show that immunotherapy, when performed appropriately and properly, not only causes a decrease in symptoms, use of medication and an improved quality of life, but also that the shift from allergic rhinitis to asthma is substantially decreased and the development of new allergies is diminished. In addition, laboratory studies continue to demonstrate significant changes in the immune system moving the patient towards a non-allergic immune function T helper type 2 cell to T helper type 1 cell shift. SUMMARY: In summary, recent research clearly continues to demonstrate the efficacy of immunotherapy not only as a therapeutic agent but as a preventive one as well. PMID- 14752339 TI - Animal models of food allergy. AB - PURPOSE OF REVIEW: The focus of this review will be on recent animal models of food allergy. Animal models are being used to investigate underlying mechanisms of IgE-mediated disease and for prophylactic/intervention therapies to treat allergic disease. RECENT FINDINGS: Considerable advances have been made in the dosage and use of sensitization routes with and without adjuvant and determinations of the pathophysiology of food allergy in murine, dog and swine food allergy models. Continued research on the neuroendocrine and novel immunoregulatory peptides is also providing new insight into inflammatory regulation and immunity. With the advent of genetically modified food crops, animal models are becoming a central theme for prediction/assessment of allergenicity for novel proteins based upon known food allergens. Therapeutic strategies involving cytokine and allergen, DNA immunizations and the use of probiotics are receiving new interest. SUMMARY: Although murine models still predominate the literature with respect to animal models of food allergy, the atopic dog and neonatal swine model are contributing knowledge with respect to symptoms more closely related to human allergic responses. Continuing investigations into the mechanisms of IgE-mediated food allergy and therapeutic strategies are providing new insights into prevention and intervention therapies for food allergy. PMID- 14752340 TI - Allergen-immunostimulatory oligodeoxynucleotide conjugate: a novel allergoid for immunotherapy. AB - PURPOSE OF REVIEW: To summarize the data of both preclinical studies and initial clinical trials of a novel allergoid for allergen specific immunotherapy. This allergoid consists of allergen covalently coupled to immunostimulatory oligodeoxynucleotide DNA sequences. RECENT FINDINGS: Recently, immunostimulatory oligodeoxynucleotide sequences, also called unmethylated cytosin-guanine dinucleotide motifs, have been discovered that act as strong T helper 1 response inducing adjuvants in mice. Although mixing allergens with immunostimulatory DNA sequences induces T helper 1 responses in T helper 2 biased mice, the allergens in such mixes could still cause anaphylactic reactions when used in humans which is one of the reasons why immunotherapy has gradually been falling out of favor. Therefore, we made allergen-immunostimulatory oligodeoxynucleotide conjugates and investigated their immunogenicity and allergenicity in animal models of allergy. These conjugates were highly immunogenic for inducing T helper 1-like antiallergen responses and reversed T helper 2 responses and symptoms of asthma in mouse models. They were also less allergenic, as shown by the reaction with human immunoglobulin E antibodies and by histamine release from basophils of allergic patients. Preliminary phase I and II trials in ragweed allergic patients showed that allergen-immunostimulatory oligodeoxynucleotide conjugates are well tolerated, less allergenic and induce immunoglobulin G antiallergen antibodies more rapidly than allergen extracts without significantly increasing the immunoglobulin E titer. SUMMARY: Allergen-immunostimulatory DNA conjugates induce T helper 1 and down regulate preexisting T helper 2 anti-allergen responses in mice. Initial phase I and II trials in ragweed allergic patients showed that ragweed allergen-DNA conjugates are well tolerated and induce a rapid immunoglobulin G but not E response. The data show that allergen-DNA conjugates are a novel type of allergoid that have great potential for a safe and potent form of allergen specific immunotherapy. PMID- 14752341 TI - Preventive capacity of subcutaneous immunotherapy. AB - PURPOSE OF REVIEW: The scope of this review is to highlight important and interesting articles about the preventive aspects and capacity of allergen specific subcutaneous injection immunotherapy. RECENT FINDINGS: There is increasing amount of data that immunotherapy may have a preventive effect in carefully selected patients, both in the development of asthma in patients with allergic rhinitis and in the development of new immunoglobulin E-mediated sensitizations. SUMMARY: The preventive capacity of subcutaneous immunotherapy in suppression of the development of new immunoglobulin E sensitizations and the progression of the allergic disease into a more severe form are interesting topics in the field of allergen-specific immunotherapy. Allergen-specific immunotherapy, the subcutaneous administration of increasing doses of allergen extracts, is the only specific and curative approach towards the treatment of immunoglobulin E-mediated allergy. Immunotherapy affects the natural course of allergic disease, and induces prolonged clinical remission when the treatment is continued for several years, which has raised the question of whether it should be considered earlier in the management of allergic diseases. There is an urgent need for large controlled multi-center studies to show the preventive aspects in different patient populations. PMID- 14752342 TI - Immunotherapy in children. AB - PURPOSE OF REVIEW: Specific allergen immunotherapy although being performed for almost 100 years, is still being addressed in clinical trials. In children the sublingual allergen application has gained considerable interest. RECENT FINDINGS: Controlled long-term trials suggest that specific allergen immunotherapy has the capacity to modify the natural history of allergic airway disease by reducing the incidence of new sensitivities non-specifically, reducing allergic symptoms years after discontinuation and preventing the incidence of asthma. The current evidence on sublingual immunotherapy is not sufficient to recommend this mode of treatment in clinical practice yet. SUMMARY: Specific allergen immunotherapy should be considered in children in whom IgE-mediated allergic symptoms cannot be adequately controlled by symptomatic treatment. PMID- 14752343 TI - The effect of amicar on perioperative blood loss in idiopathic scoliosis: the results of a prospective, randomized double-blind study. AB - STUDY DESIGN: A prospective, randomized, double-blind Institutional Review Board approved study evaluating the efficacy of Amicar (epsilon aminocaproic acid), an antifibrinolytic agent, in decreasing perioperative blood loss in idiopathic scoliosis. OBJECTIVES: To compare the perioperative (intraoperative and postoperative) blood loss and the need for autologous and homologous blood replacement in two groups of essentially identical patients undergoing a posterior spinal fusion for idiopathic scoliosis. SUMMARY OF BACKGROUND DATA: Reducing perioperative blood loss and the need for transfusion in patients undergoing spinal surgery is important to orthopedic surgeons. Recently, there has been interest in pharmacologic agents, particularly Amicar and Aprotinin, to assist in decreasing perioperative blood loss. In 2001, in a preliminary study, we demonstrated that Amicar appeared to be effective in reducing perioperative blood loss in patients with idiopathic scoliosis undergoing a posterior spinal fusion and segmental spinal instrumentation. This was a study of 28 consecutive patients receiving Amicar compared to a historical control group of the 31 previous consecutive patients with the same study criteria. The current study was performed to confirm our preliminary findings. METHODS: We analyzed the perioperative blood loss of 36 patients with idiopathic scoliosis who were blindly randomized by the operating room pharmacy into an Amicar and control group. The criteria to be included in the study was the same as the preliminary study: diagnosis of idiopathic scoliosis, age at surgery 11 to 18 years, posterior spinal fusion and segmental spinal instrumentation only, autogenous iliac crest bone graft or homologous cancellous bone graft, and a signed agreement to participate in the study. The patients in both groups had the same anesthetic technique, intraoperative procedure, instrumentation, postoperative management, and standardized indications for transfusions. RESULTS: Before surgery, the patients in both groups were essentially identical. The distribution of patients and their results was not known until the completion of the study. Patients in the Amicar group demonstrated a statistically significant decrease in perioperative blood loss and the need for autologous blood transfusion. Interestingly, this decrease was predominantly in the postoperative suction drainage. This may be due to elevated fibrinogen levels induced by Amicar. The patients taking Amicar had no intraoperative or postoperative thromboembolic complications. CONCLUSIONS: The results of this study confirmed that the use of intraoperative Amicar is a safe, effective, and inexpensive method to significantly reduce perioperative blood loss in patients with idiopathic scoliosis undergoing posterior spinal fusion and segmental spinal instrumentation. The results have allowed us to reduce our recommendation for perioperative autologous blood donation, thereby further decreasing costs. PMID- 14752344 TI - Thoracic pedicle morphometry in vertebrae from scoliotic spines. AB - STUDY DESIGN: A morphometric analysis of thoracic pedicles in vertebrae from scoliotic specimens. OBJECTIVE: The objective of this study was to quantify the changes occurring in thoracic pedicles affected by a scoliotic deformity. SUMMARY OF BACKGROUND DATA: There exists a lot of controversy in the literature concerning the shape and size of thoracic pedicles in idiopathic scoliosis. In recent years, thoracic pedicle screws are being used more frequently in corrective spine surgery, but few studies have evaluated the morphology of scoliotic thoracic pedicles. MATERIAL AND METHOD: Thirty scoliotic specimens with curves presenting various degrees of severity were studied using a three dimensional digitizing protocol developed to create a precise three-dimensional reconstruction of the vertebrae. Twenty-two parameters describing specifically the pedicles were then calculated for each vertebra from these reconstructions. Every scoliotic specimen was then matched with a normal specimen to provide for a representative control group and comparisons were made on pedicle width, length, height, surface, and orientation. RESULTS: A total of 683 thoracic vertebrae were measured (325 scoliotic and 358 normal vertebrae). Pedicles located on the concavity of typical right thoracic curves were found to be significantly thinner than their normal counterparts with a maximal mean difference of 1.37 mm at T8. The pedicles on the concavity of the high thoracic compensatory curve were also found to be significantly diminished with a maximal mean difference of 1.68 mm at T4. Mean left pedicle width at T8 (concavity) and mean right pedicle width at T4 (concavity) were found to be 4.08 mm and 2.60 mm, respectively. Pedicle length was found to be slightly increased, and pedicle height was found to be slightly decreased in pedicles from scoliotic spines with no preference for concavity or convexity. Pedicle orientation and inclination were unchanged with respect to each corresponding vertebral body. CONCLUSION: These results are of critical importance for clinicians performing spinal corrective surgery in patients with AIS. Pedicle width is significantly diminished on the concavity of scoliotic curves. Our results advocate caution in the use of pedicle screws in the thoracic spine especially on the concave side of the curve. PMID- 14752345 TI - Efficacy of osteogenic protein-1 in a challenging multilevel fusion model. AB - STUDY DESIGN: A therapeutic study compared the influence of osteogenic protein-1 to autograft and collagen carrier in multilevel sheep spine fusions. OBJECTIVE: To evaluate the efficacy of osteogenic protein-1 compared to autograft and collagen carrier in achieving fusion in a challenging multilevel lumbar spine ovine model. SUMMARY OF BACKGROUND DATA: Bone morphogenetic proteins can successfully augment spinal fusion. To date, all the preclinical and clinical studies using bone morphogenetic proteins have evaluated single-level fusion. In practice, multiple level fusions are commonly required for various conditions, like spinal deformity. METHODS: Eighteen sheep underwent three-level spine fusion. Six sheep were treated with osteogenic protein-1 and its carrier, autograft, or with the carrier alone. Specimens were analyzed for evidence of fusion by palpation, radiographic and histologic analysis, and biomechanical testing. RESULTS: Manual palpation testing for the presence of fusion showed none of the specimens fused all three levels or fused at the lumbosacral junction. No statistically significant difference was found between the osteogenic protein-1 and autograft groups' fusion rates based on radiographic grading (P = 0.65) or biomechanical testing. Histologic analysis showed no qualitative difference in bone morphology or cellularity of fusion masses when comparing the autograft and osteogenic protein-1 specimens. CONCLUSIONS: No model before this exists that tests the efficacy of bone morphogenic proteins in as challenging an environment. Extrapolation of single-level preclinical and clinical studies with bone morphogenic proteins for use in multilevel fusion requires careful review. Autograft and osteogenic protein-1 had similar rates of fusion. A high rate of nonunion is seen with this multiple level fusion to the sacrum using autograft or osteogenic protein-1. The biologic enhancement with osteogenic protein-1 is not able to overcome this mechanically rigorous model. PMID- 14752347 TI - Validity and internal consistency of a whiplash-specific disability measure. AB - STUDY DESIGN: Cross-sectional study of patients with whiplash-associated disorders investigating the internal consistency, factor structure, response rates, and presence of floor and ceiling effects of the Whiplash Disability Questionnaire (WDQ). OBJECTIVES: The aim of this study was to confirm the appropriateness of the proposed WDQ items. SUMMARY OF BACKGROUND DATA: Whiplash injuries are a common cause of pain and disability after motor vehicle accidents. Neck disability questionnaires are often used in whiplash studies to assess neck pain but lack content validity for patients with whiplash-associated disorders. The newly developed WDQ measures functional limitations associated with whiplash injury and was designed after interviews with 83 patients with whiplash in a previous study. METHODS: Researchers sought expert opinion on items of the WDQ, and items were then tested on a clinical whiplash population. Data were inspected to determine floor and ceiling effects, response rates, factor structure, and internal consistency. Packages of questionnaires were distributed to 55 clinicians, whose patients with whiplash completed and returned 101 questionnaires to researchers. RESULTS: No substantial floor or ceiling effects were identified on inspection of data. The overall floor effect was 12%, and the overall ceiling effect was 4%. Principal component analysis identified one broad factor that accounted for 65% of the variance in responses. Internal consistency was high; Cronbach's alpha = 0.96. CONCLUSIONS: Results of the study supported the retention of the 13 proposed items in a whiplash-specific disability questionnaire. Dependent on the results of further psychometric testing, the WDQ is likely to be an appropriate outcome measure for patients with whiplash. PMID- 14752348 TI - The posterior approach for lumbar and thoracolumbar adolescent idiopathic scoliosis: posterior shortening and pedicle screws. AB - STUDY DESIGN: Prospective clinical case series. BACKGROUND DATA: Lumbar and thoracolumbar adolescent idiopathic scoliosis has traditionally been treated with an anterior approach and instrumentation. This anterior method often has had problems with kyphosis, pseudarthrosis, and loss of correction. The senior author has had good results treating these same lumbar and thoracolumbar curves posteriorly with wide posterior release and segmental instrumentation. In this series of his evolving technique, he adds pedicle screws as the sole anchor in the thoracolumbar/lumbar curves. OBJECTIVES: To prospectively evaluate outcomes, coronal and sagittal radiographic results, balance parameters, complications, and reoperations in a group of consecutive patients with lumbar and thoracolumbar adolescent idiopathic scoliosis. These patients were surgically treated with wide posterior release and segmental posterior screw instrumentation with 2-year minimum follow-up (range 26-47 months). METHODS: Sixty-two consecutive patients with thoracolumbar and lumbar adolescent idiopathic scoliosis were treated with a wide posterior release and segmental pedicle screw instrumentation limited to the curve defined by the Cobb measurement. The patients were evaluated clinically and radiographically at intervals up to 36 months. There was 2-year minimum follow up. RESULTS: One patient was lost to follow-up. Of the remaining 61 patients, there were 51 Lenke 5 Type curves, 7 Lenke Type 3C curves, and 3 Lenke Type 6 curves. Only the curve defined by the Cobb measurement was fused. A total of 613 pedicle screws were placed safely. Average coronal correction of the thoracolumbar/lumbar curves was from 52 degrees to 10 degrees (80%). In the sagittal plane, lumbar lordosis was normalized from 41 degrees with a wide range (20 degrees -70 degrees ) to 42 degrees with a normal range (34 degrees -47 degrees ). There were no pseudoarthroses, no reoperations, no infections, no problems with screw placement, and excellent maintenance of correction at last follow-up. The lowest instrumented vertebrae had 81% correction of coronal angulation, center sacral line to lowest instrumented vertebrae was improved from 2.4 cm to 0.7 cm, and apex to center sacral line was improved from 5.2 cm to 1.5 cm. The C7 plumb line to center sacral line was also improved from 2.5 cm to 0.6 cm, illustrating the centering of the trunk. CONCLUSIONS: Wide posterior release and segmental pedicle screw instrumentation has excellent radiographic and clinical results with minimal complications. There were no pseudoarthroses and no reoperations. PMID- 14752346 TI - Biomechanical evaluation of contemporary posterior spinal internal fixation configurations in an unstable burst-fracture calf spine model: special references of hook configurations and pedicle screws. AB - STUDY DESIGN: This study attempts to determine the most biomechanically rigid posterior spinal instrumentation configuration in a burst-fracture calf spine model. OBJECTIVES: To compare the biomechanical stability of contemporary posterior spinal instrumentation in various hook and screw configurations in an unstable calf spine model. SUMMARY OF BACKGROUND DATA: Burst-fractures are relatively common injuries seen in the setting of spinal trauma. The use of posterior-only configurations in the treatment of this deformity has become a much more popular approach because of the relative ease of applying the instrumentation. METHODS: Fresh frozen in vitro study using 10 calf spines involving the T11-S1 vertebral segments. Pure moment forces including flexion, extension, axial rotation, and lateral bending were applied to the top of the spinal column at T11. Testing was first performed on all intact specimens. A corpectomy was then performed at L2. Testing was then repeated on each of the ten specimens after internal fixation with different posterior spinal configurations using ISOLA instrumentation (DePuy AcroMed Inc., Raynham, MA). RESULTS: With regards to flexion-extension and lateral bending, all configurations except for distraction hook-rod construct provided stability greater than the intact spine. The distraction hook-rod configuration failed to control extension (P > 0.05) above the intact specimen. All pedicle screw constructs were more rigid than the hook-rod constructs in axial rotation at the level of injury (P < 0.001). CONCLUSIONS: The motion segment at the corpectomy site is adequately stabilized by contemporary spinal internal fixation configurations tested except for the distraction-hook stabilization. Axial rotation is generally poorly controlled by posterior-only internal fixation. Pedicle screw instrumentation was the most rigid compared with other forms of stabilization in stabilizing a burst corpectomy defect. Based on this study, pedicle screw configurations are preferred over hook-rod strategies in the posterior stabilization of a burst corpectomy anterior defect. Among hook-rod configurations, the distraction hook rod strategy provided the least stability. PMID- 14752350 TI - Can we predict the ultimate lumbar curve in adolescent idiopathic scoliosis patients undergoing a selective fusion with undercorrection of the thoracic curve? AB - STUDY DESIGN: Retrospective review of anterior and posterior fusions for treatment of adolescent idiopathic thoracic scoliosis. OBJECTIVES: To delineate the best factors determining final lumbar curve magnitude in patients with adolescent idiopathic scoliosis undergoing a selective thoracic anterior or posterior spinal fusion at or proximal to the first lumbar vertebra. SUMMARY OF BACKGROUND DATA: Although spontaneous lumbar curve correction occurs consistently following a selective thoracic anterior or posterior spinal fusion, the degree of correction is somewhat unpredictable. METHODS: One hundred consecutive patients with major thoracic-compensatory lumbar adolescent idiopathic scoliosis treated by a single surgeon with either selective posterior spinal fusion (n = 44) or anterior spinal fusion (n = 56) of the main thoracic region with an unfused lumbar spine with a lumbar B modifier (lumbar apex touching the center sacral vertical line) or lumbar C modifier (lumbar apex completely lateral to the center sacral vertical line) were retrospectively reviewed. RESULTS: Those patients who maintained excellent postoperative coronal balance, with spontaneous lumbar curve correction, had their thoracic Cobb corrected intraoperatively to a measurement very close to but not more than that of the preoperative thoracic push-prone Cobb. Stepwise multiple linear regression analysis was used to develop a formula to help predict lumbar response in those patients undergoing selective thoracic fusion. This is represented in the following formula: Final lumbar Cobb = 14.4 + 3.06 (lumbar modifier; 0 = B, 1 = C) + 0.30 (preoperative standing lumbar Cobb) - 0.18 (preoperative supine lower Cobb) + 0.81(preoperative push/prone lumbar Cobb) - 0.15(preoperative standing thoracic Cobb) - 0.16(% thoracic Cobb change from preoperative to immediate postoperative). Final model R2 = 0.72. CONCLUSIONS: Of the preoperative measurements examined, the preoperative push-prone is the best preoperative flexibility radiograph to predict the final lumbar curve measurement and, along with other factors, can be used to formulate a model that will help the treating surgeon more confidently predict the final lumbar curve response in patients undergoing a selective thoracic fusion. PMID- 14752351 TI - Predictors of progression of scoliosis after decompression of an Arnold Chiari I malformation. AB - STUDY DESIGN: Retrospective study of patients with scoliosis and an Arnold Chiari I malformation requiring operative management. OBJECTIVES: Determine the factors that could predict whether a particular spinal deformity might progress despite neurosurgical management of Arnold Chiari I malformation. SUMMARY OF BACKGROUND DATA: Few studies have documented the relationship between diagnosis and treatment of Arnold Chiari I malformation and associated spinal deformities. Most studies mix neural axis abnormalities and contain limited information about the spinal deformity. METHODS: Medical records, radiographs, and magnetic resonance images of patients were evaluated focusing on age and findings at presentation, characteristics of presenting and follow-up spinal deformities, and the specifics of neurosurgical and orthopedic management. Patients were divided into two groups: those whose curves progressed >10 degrees or to surgical range (largest curve >45 degrees ) after neurosurgical intervention (progressors) and those whose curves stabilized or decreased (nonprogressors). RESULTS: Eight progressors presented at an average age of 11.4 years (range 2-19) and were followed for 6.3 years (range 2-15). Seven nonprogressors presented at 6.5 years of age (range 5 10) and were followed for 6.6 years (range 3.5-14). Neurosurgical procedures were equivalent in both groups; however, surgical revisions were required in 3 out of 8 progressors and 1 out of 7 nonprogressors. All progressors had a double scoliosis curve; but only one nonprogressor had a double curve. Six out of 8 progressors and 0 out of 7 nonprogressors had a rotation >or=2+ and 50% of progressors had a thoracic kyphosis >50 degrees compared to 1 out of 7 nonprogressors. CONCLUSIONS: In this series, progression of spinal deformity after neurosurgical management of Arnold Chiari I malformation was associated with later age at neurosurgical decompressions and initial neurologic symptoms, double scoliosis curve patterns, kyphosis, rotation, and larger curve at presentation. PMID- 14752352 TI - Noninvasive magnetic resonance to three-dimensional rotational x-ray registration of vertebral bodies for image-guided spine surgery. AB - STUDY DESIGN: Magnetic resonance (MR) and three-dimensional rotational x-ray (3DRX) images of cadaveric spinal segments were registered using a conventional point-based technique and a noninvasive technique called maximization of mutual information (MMI). OBJECTIVES: To assess the feasibility and accuracy of MMI based registration in comparison with point-based registration as a new noninvasive image-to-patient registration technique for use in minimally invasive image-guided spine surgery. SUMMARY OF BACKGROUND DATA: In image-guided orthopedic surgery, correspondence between before surgery acquired images and the patient is required. Currently, this necessitates an invasive registration step, in which anatomic landmarks on the bone surface have to be physically touched by a pointer. To overcome this invasive procedure, we propose using a calibrated 3DRX system, a modality that can visualize high contrast objects intraoperatively and that provides a direct correspondence between the image data and the patient. A noninvasive voxel-based technique is used to register the intraoperative 3DRX image to a before surgery acquired MR image. METHODS: Cadaveric thoracic and lumbar spine segments were implanted with markers, which were used as landmarks. The accuracy of the landmark-based registration was compared with MMI-based registration using the residual errors on the marker positions. RESULTS: The errors made using the point-based registration technique were compared with the errors made with MMI. The results show a statistically significant lower error (P < 0.01) for the proposed MMI method. CONCLUSIONS: Noninvasive MMI registration of intraoperative 3DRX images to preoperative MR images is more accurate than point based registration in cadaveric spine segments. It is therefore a promising technique for replacing the invasive landmark-based registration that is currently used in image-guided spine surgery. PMID- 14752353 TI - An analysis of chest wall and diaphragm motions in patients with idiopathic scoliosis using dynamic breathing MRI. AB - STUDY DESIGN: The motions of chest wall and diaphragm were measured in patients with scoliosis and healthy individuals using dynamic breathing magnetic resonance imaging. OBJECTIVES: To investigate the motions of the chest wall and the diaphragm during deep breathing in patients with idiopathic scoliosis. SUMMARY OF BACKGROUND DATA: Despite the central role of the respiratory motions in maintaining ventilation, it is difficult to measure and describe the motions because of the limited methods available. The kinematics of respiration in scoliosis have not yet been clarified. METHODS: Dynamic fast spoiled gradient recalled echo sequences were used for 18 patients with idiopathic scoliosis and 9 healthy individuals. The chest wall and diaphragm motions were evaluated using a cineloop view and a fusion display of maximal inspiratory and expiratory images. The data were analyzed quantitatively by measuring displacements. RESULTS: Respiratory chest wall movements were significantly restricted in patients with scoliosis, although the diaphragm motion was normal. An analysis of the fusion magnetic resonance displays superimposed with the maximal inspiratory and expiratory images facilitated the understanding of the abnormal respiratory motion in patients with scoliosis. There was some correlation between the values of respiratory motions and pulmonary function tests. CONCLUSIONS: The present technique is useful for assessing respiratory mechanisms dynamically and noninvasively. One of the distinguishing features of respiratory dysfunction in patients with scoliosis is their limited chest wall motion. PMID- 14752354 TI - Comparison of SRS questionnaire results submitted by both parents and patients in the operative treatment of idiopathic scoliosis. AB - STUDY DESIGN: A prospective, cross-sectional analysis of patients with operative idiopathic scoliosis comparing Scoliosis Research Society's Outcomes Instrument (SRS-24) scores from both parents and patients obtained separately on the same day along with pertinent radiographic data. OBJECTIVE: To determine the correlation of parents' and patients' perspectives of the patients' preoperative and postoperative experience using the SRS-24 questionnaire emphasizing parent patient disparities. SUMMARY OF BACKGROUND DATA: No report to our knowledge has addressed the association between parent assessments of their child's presurgical and postsurgical outcome as verified by SRS-24 questionnaire data. MATERIALS AND METHODS: A total of 101 patients with operative idiopathic scoliosis were analyzed based on paired parent-patient SRS-24 data and radiographs performed on the same day. Patients were analyzed by age at date of examination (9-23 years, mean 17 +/- 5 years), gender (16 male, 85 female), major curve magnitude (41-126 degrees ), procedure type [preoperative (22 pairs), anterior spinal fusion (49 pairs), anterior spinal fusion/posterior spinal fusion (19 pairs), posterior spinal fusion (46 pairs)], and time from surgery (preoperative, postoperative 1 93 months, mean 24 months). All questionnaire scores were classified based on domains of pain, self-image, function, overall satisfaction, and total score. RESULTS: In overall time periods, parents consistently scored higher than their children in the self-image (P = 0.0001), satisfaction (P = 0.0001), and total score (P = 0.04), but not pain or function. Before surgery, parents overestimated patients' scores in self-image (P = 0.002) by 7.5% but not other domains. Satisfaction differences (P = 0.04) improved with increasing age of the patient, but not other domains. There were no significant differences with gender or preoperative/postoperative major curve magnitudes. CONCLUSION: Based on SRS-24 data, parents typically scored higher than their children in the operative treatment of idiopathic scoliosis in total score, self-image, and overall satisfaction. Some parent-patient scores correlated better with increasing age of the patient, and later in the postoperative period. PMID- 14752355 TI - Force production parameters in patients with low back pain and healthy control study participants. AB - STUDY DESIGN: A control group study with repeated measures. OBJECTIVE: To compare isometric force production parameters in low back pain and healthy study participants. SUMMARY AND BACKGROUND DATA: Recent evidence suggests that chronic patients with low back pain exhibit deficits in trunk proprioception and motor control. The control of force and its between-trial variability are often taken as critical determinants of performance. We compared various force time characteristics in patients with low back pain and healthy study participants. METHODS: Fifteen control study participants and 16 patients with low back pain participated in this study. Study participants were required to exert 50% and 75% of the maximal trunk flexion and extension. In a learning phase, visual and verbal feedback was provided. Following these learning trials, study participants were asked to perform 10 trials without any feedback. Time to peak force, time to peak force variability, peak force variability, and absolute error in peak force were calculated. Time to peak and peak dF/dt were computed to determine if the first peak of dF/dt could predict the peak force achieved. RESULTS: Two subgroups of patients with low back pain were identified. Controls and patients with low back pain with more pain showed faster time to peak force than patients with low back pain with less pain (331 ms and 341 ms vs. 574 ms, respectively). Linear regressions showed that, for control study participants and low back pain study participants with more pain, peak dF/dt explained 94.0% and 97.0% of the variance observed in peak force while 84.4% was explained for low back pain study participants with less pain. Peak force variability and absolute error in peak force were similar for all groups. CONCLUSIONS: Patients with low back pain were able to produce isometric forces with an accuracy similar to control study participants. The longer time to peak force and the smaller percentage of variance observed for the linear regressions suggest that some patients with low back pain adopted a control mode that was less "open-loop." It is possible that this mode of producing forces results from an adaptation to chronic pain or tissue degeneration. PMID- 14752356 TI - Late complications of adult idiopathic scoliosis primary fusions to L4 and above: the effect of age and distal fusion level. AB - STUDY DESIGN A retrospective analysis of primary cases of adult idiopathic scoliosis treated with long instrumented fusions from the thoracic spine proximally to segments that range from T11 to L4 distally. OBJECTIVE: To analyze whether patients requiring revision surgery had lower postoperative SRS-24 scores; age >or=40 years correlated with higher rates of revision surgery; disc degeneration below the fusion occurred more commonly with a more distal lowest instrumented vertebra or older patient age (>or=40 years); and whether smokers had higher rates of major complications or revision surgery. SUMMARY OF BACKGROUND DATA: Few reports describe complications related to primary long fusions using modern 2+ rods, hook/pedicle screw instrumentation methods in the treatment of adult idiopathic scoliosis. METHODS: Sixty-seven patients were analyzed with an average age of 38.8 years (range 21-61 years). The average clinical follow-up was 7.8 years (range 2-16 years): 42 patients had >5 years follow-up, including 23 patients with >10 years follow-up. Patients were categorized by age (< or >or=40 years) and level of the lowest instrumented vertebra (T11-L2 vs. L3-L4). Upright radiographs and postoperative SRS-24 questionnaires from the latest follow-up date were analyzed. RESULTS: Patients requiring revision surgery had lower total score (average 72.0) than those that did not (total score = 94.2; P = 0.01). More specifically, patients with pseudarthrosis had lower total scores (average 74.7) than those without (average total score = 93.5; P = 0.02). When analyzing age, there were similar rates of pseudarthrosis, but higher rates of transition syndrome (2) and sagittal/coronal imbalance (1 each) in patients >or=40 years. Subsequent distal disc degeneration did not correlate significantly with more distal lowest instrumented vertebra or older patient age. Smokers did not have higher rates of major complications or revision surgery than nonsmokers. CONCLUSIONS: Patients with adult idiopathic scoliosis and long fusions had similar pseudarthrosis rates, but higher rates of transition syndrome when lowest instrumented vertebra was L3-L4 relative to levels T11-L2. When categorized by age, complication rates were similar in each group. Patients with pseudarthroses or other diagnoses requiring revision surgery had lower SRS-24 total scores than those without (P = 0.02 and P = 0.01, respectively). PMID- 14752357 TI - Treatment of pyogenic vertebral osteomyelitis with anterior debridement and fusion followed by delayed posterior spinal fusion. AB - STUDY DESIGN: Case series. OBJECTIVES: The results of surgical treatment of osteomyelitis with anterior debridement and fusion followed by delayed posterior stabilization and fusion are presented. METHODS: Forty-two patients with vertebral osteomyelitis are presented. There were 5 cervical, 12 thoracic, 1 thoracolumbar, 19 lumbar, and 5 lumbosacral lesions. The most frequently identified organism was Staphylococcus aureus. Most patients had significant comorbidities, including diabetes, or were immunocompromised. Ninety percent had elevated erythrocyte sedimentation rates and C-reactive proteins, while white blood cell counts were less reliably elevated. Imaging studies included radiographs, CT scans, and MRIs. All patients were treated with anterior debridement and strut grafting followed by 14.4 days of intravenous antibiotics and delayed instrumented posterior fusions and received 6 weeks of intravenous antibiotics after surgery. RESULTS: All patients had resolution of their infections with no recurrence. There were two deaths. Neurologic deficits resolved in all patients. The diagnosis of pyogenic vertebral osteomyelitis is frequently delayed and presents a significant surgical challenge. The indications for surgical debridement were neurologic compromise, failed medical treatment, soft tissue extension, extensive vertebral body and disc space destruction, and progressive deformity. Many of these patients were severely ill at presentation and required urgent treatment. Anterior debridement and fusion followed by intravenous antibiotics allows for restoration of anterior column support and control of the infection before posterior instrumentation and fusion. CONCLUSION: This study demonstrates that anterior surgical debridement with fusion, followed by a period of intravenous antibiotics and delayed instrumented posterior fusion, is highly effective in the treatment of pyogenic osteomyelitis that has failed medical management. PMID- 14752359 TI - Free hand pedicle screw placement in the thoracic spine: is it safe? AB - STUDY DESIGN: A retrospective study. OBJECTIVE: To evaluate the safety of a free hand technique of pedicle screw placement in the thoracic spine at a single institution over a 10-year experience. SUMMARY OF BACKGROUND DATA: Thoracic pedicle screw fixation techniques are still controversial for thoracic deformities because of possible complications including neurologic. MATERIALS AND METHODS: Three hundred ninety-four consecutive patients who underwent posterior stabilization utilizing 3204 transpedicular thoracic screws by 2 surgeons from 1992 to 2002 were analyzed. The mean age was 27 + 10 years (range 5 + 3-87 + 0 years) at the time of surgery. Etiologic diagnoses were: scoliosis in 273, kyphosis in 53, other spinal disease in 68. Pedicle screws were inserted using a free hand technique similar to that used in the lumbar spine in which anatomic landmarks and specific entry sites were used to guide the surgeon. A 2-mm tip pedicle probe was carefully advanced free hand down the pedicle into the body. Careful palpation of all bony borders (floor and four pedicle walls) was performed before and after tapping. Next, the screw was placed, followed by neurophysiologic (screw stimulation with rectus abdominus muscle recording) and radiographic (anteroposterior and lateral) confirmation. An independent spine surgeon using medical records and roentgenograms taken during treatment and follow-up reviewed all the patients. RESULTS: The number of the screws inserted at each level were as follows (total n = 3204): T1, n = 13; T2, n = 60; T3, n = 192; T4, n = 275; T5, n = 279; T6, n = 240; T7, n = 230; T8, n = 253; T9, n = 259; T10, n = 341; T11, n = 488; T12, n = 572. Five hundred seventy-seven screws inserted into the deformed thoracic spine were randomly evaluated by thoracic computed tomography scan to assess for screw position. Thirty-six screws (6.2%) were inserted with moderate cortical perforation, which meant the central line of the pedicle screw was out of the outer cortex of the pedicle wall and included 10 screws (1.7%) that violated the medial wall. There were no screws (out of the entire study group of 3204) with any neurologic, vascular, or visceral complications with up to 10 years follow-up. CONCLUSIONS: The free hand technique of thoracic pedicle screw placement performed in a step-wise, consistent, and compulsive manner is an accurate, reliable, and safe method of insertion to treat a variety of spinal disorders, including spinal deformity. PMID- 14752361 TI - Direct vertebral rotation: a new technique of three-dimensional deformity correction with segmental pedicle screw fixation in adolescent idiopathic scoliosis. AB - STUDY DESIGN: A prospective study. OBJECTIVES: To introduce a new technique, direct vertebral rotation, and to compare the surgical results of direct vertebral rotation with those of simple rod derotation. SUMMARY OF BACKGROUND DATA: Pedicle screw fixation with a simple rod derotation maneuver enables a powerful coronal and sagittal plane correction in scoliosis surgery. However, the ability of achieving rotational correction is still unclear. METHODS: Thirty eight adolescent idiopathic scoliosis patients treated with segmental pedicle screw fixation were analyzed. The first group (n = 17) was treated by direct vertebral rotation; the second group (n = 21) was treated by simple rod derotation. All patients had a minimum follow-up of 2 years. Having similar preoperative curve patterns, both groups were evaluated for the deformity correction, lower instrumented vertebral tilt, and spinal balance. Apical vertebral rotation was evaluated by computed tomography scans. Surgical techniques of direct vertebral rotation were as follows: a precontoured rod was inserted into segmental screws on the concave side in thoracic scoliosis; a simple rod derotation was performed; and then the screws on the juxta-apical vertebrae, both on concave and convex sides, were rotated opposite direction to the rod derotation. Then, all the screws were sequentially tightened. RESULTS: In the direct vertebral rotation group, the average preoperative apical vertebral rotation of 16.7 degrees was corrected to 9.6 degrees, showing 42.5% correction, whereas in the simple rod derotation group, the correction was negligible from 16.1 degrees to 15.7 degrees (2.4%). In the direct vertebral rotation group, the average preoperative thoracic curve of 55 degrees was corrected to 12 degrees (79.6%), and the lumbar curve of 39 degrees was corrected to 7 degrees (80.5%). In the simple rod derotation group, the preoperative thoracic curve of 53 degrees was corrected to 17 degrees (68.9%), and the lumbar curve of 39 degrees was corrected to 16 degrees (62.2%). The average lower instrumented vertebral tilt correction was 80.6% and 66.3% in the directvertebral rotation and the simple rod derotation group, respectively. There were statistically significant differences in the coronal curve, lower instrumented vertebral tilt, and rotational correction (P < 0.05, Mann-Whitney U test). Thoracic kyphosis was improved in both groups. CONCLUSIONS: Segmental pedicle screw fixation with "direct vertebral rotation" showed better rotational and coronal correction than "simple rod derotation." PMID- 14752362 TI - Contributions of flexion-extension cyclic loads to the lumbar spinal segment stability following different discectomy procedures. AB - STUDY DESIGN: An in vitro biomechanical cadaveric study. OBJECTIVES To elucidate the effect of flexion-extension cyclic loads on the motion behavior of lumbar spine after different discectomies. SUMMARY OF BACKGROUND DATA: Biomechanical cadaveric studies to evaluate the effect of discectomy have been performed and have indicated the relevance between the volume of removed disc materials and increase of motion in affected intervertebral disc. However, there are no biomechanical studies to investigate the motion behavior of injured intervertebral disc after cyclic loads. METHODS: Twenty-eight lumbar functional spinal units were randomized into fenestration, annulotomy, limited discectomy, and radical discectomy groups. Pure bending moments were applied to simulate various loading modes and determine the resulting displacements before and after surgery, and after cyclic loads of 1,000, 5,000, and 10,000 cycles at a frequency of 0.5 Hz and a force of +/-3.0 Nm. Change of range of motion (ROM) was compared among each group. RESULTS: Following surgery, in the radical discectomy group, the relative change of ROM mostly increased in all motion directions except right lateral bending. On the other hand, during cyclic loads up to 10,000 cycles, in the limited discectomy group, the relative change of ROM mostly increased in all motion directions except right lateral bending. CONCLUSIONS: These results demonstrate that the effect of cyclic loads after discectomy may increase ROM, leading to spinal instability even if the increase in ROM does not occur immediately after surgery for the minimum removal of nucleus pulposus case. Clinically, this may underscore the importance of postoperative lumbar support. PMID- 14752363 TI - The possibility to use simple validated questionnaires to predict long-term health problems after whiplash injury. AB - STUDY DESIGN: A prospective follow-up study. OBJECTIVES: To evaluate the relation of the state of health before the accident and the significance of the symptoms reported soon after the injury to the situation 3 years after the injury. To evaluate the possibility of using simple validated questionnaires to predict long term health problems after the injury. SUMMARY OF BACKGROUND DATA: A whiplash injury is generally benign in its natural course. However, some of the patients have diverse and prolonged symptoms. Although several prognostic factors have been suggested for the poor recovery, in most cases the factors leading to prolonged disability remain unclear. METHODS: In collaboration with traffic insurance companies, we gathered information of neck injuries that occurred in traffic accidents in Finland in 1998. After the insurance company received a notification of a neck injury and consent from the injured party to participate in the study, they sent the information to the research team. The first inquiry was sent to the patients as soon as possible. One- and three-year follow-up questionnaires were posted to those who responded to the first inquiry. A total of 144 persons returned the 3-year follow-up questionnaires and form the material of this study. RESULTS A poor state of health or frequent neck pain or headache before the accident did not have any significant relation to the poor outcome 3 years after the accident. The extent of neck pain and lower back pain reported soon after the accident was significantly associated to a poor outcome in the follow-up. The Neck Disability Index questionnaire was significantly related to the poor outcome after 3 years. CONCLUSIONS: The subjective experience of a notably decreased level of activity because of the neck pain when supplemented by the enhanced score of Neck Disability Index questionnaire predicts well poor outcome in long-term follow-up and can be used as a tool to identify persons who are at risk to suffer long-term health problems after whiplash injury. PMID- 14752364 TI - Congenital scoliosis due to unilateral unsegmented bar: posterior spine fusion at age 12 months with 44-year follow-up. AB - STUDY DESIGN: Retrospective case report. PURPOSE: Long-term follow-up. METHODS: A 44-year follow-up after surgery at the age of 1 year for congenital scoliosis with original and current information. BACKGROUND INFORMATION: This long a follow up after so young a fusion has not previously been reported. Shorter follow-ups have not revealed the problems this patient has had subsequently. RESULTS: Low back pain began 22 years after surgery (age 23 years) and cervical pain began 24 years after surgery (age 25 years). Anterior cervical discectomy and fusion plus posterior fusion of two disc levels were necessary at the age of 36 years. Continued low back pain resulting from multilevel degeneration causes major disability. CONCLUSIONS: This patient had an excellent result from his posterior spine fusion at the age of 1 year, with no loss of curve correction, no "crankshafting," and minimal loss of vital capacity. Long-term follow-up did indicate major disabilities resulting from both cervical and lumbar degenerative disc disease. PMID- 14752365 TI - Surgical management of dissociated motor loss following complex cervical spine reconstruction. AB - STUDY DESIGN: A case of surgical management of dissociated motor loss after decompression of the cervical spine is reported. OBJECTIVES: To present a rationale for surgical treatment of postdecompressive cervical radiculopathy with an illustrative case example. SUMMARY OF BACKGROUND DATA: The unusual complication of radiculopathy after multilevel cervical decompressive procedures is characterized by pain or dissociated motor weakness of the C5 and C6 nerve roots. Conservative management paradigms, including analgesics and steroids, are the rule, but symptoms often persist for many months. There are currently no reports describing foraminotomy as a means of more rapidly alleviating the symptoms of radicular pain and deltoid and biceps brachii weakness seen in the postoperative setting. METHODS: We present a case of bilateral C5 and C6 radiculopathy following multilevel cervical decompression for cervical spondylotic myelopathy, which we treated with posterior foraminotomies. RESULTS: The patient reported complete resolution of his dermatomal pain and demonstrated rapid improvement in upper extremity strength as compared to traditional conservative treatments. The historical experience and pathogenesis regarding this postoperative complication are reviewed. The rationale of root-specific posterior decompression for this debilitating complication is discussed. CONCLUSIONS: Foraminal decompression of the affected nerve roots as demonstrated here has not been described for postdecompressive dissociated motor loss. Such an approach may offer earlier and more complete relief to patients suffering from this unfortunate complication. PMID- 14752366 TI - Traumatic anterior atlantoaxial subluxation occurring in a professional rugby athlete: case report and review of literature related to atlantoaxial injuries in sports activities. AB - STUDY DESIGN: This is a case report of traumatic anterior atlantoaxial subluxation occurring in a professional rugby athlete with a literature review. OBJECTIVE: To report a rare case of traumatic anterior atlantoaxial subluxation in a professional rugby athlete. SUMMARY OF BACKGROUND DATA: Atlantoaxial subluxation occurring in a professional athlete is very rare, and only two cases have been reported so far. METHODS: A 26-year-old professional rugby player sustained traumatic anterior atlantoaxial subluxation. The pathomechanism of the injury was hyperflexion. No quadriplegia existed, but severe anterior subluxation was found on functional radiographs and kinematic magnetic resonance imaging. RESULTS: Surgical fixation of C1-C2 was carried out, and no impairment was seen in his daily living, although he did not return to professional rugby. CONCLUSIONS: Anterior atlantoaxial subluxation in sports activities is very rare. The mechanism of this injury is hyperflexion, and the rupture of the transverse ligament is essential. Good prognosis has been reported; however, a return to high-level sports activities is thought to be a contraindication. PMID- 14752367 TI - [ISAT: a randomised trial to arbitrate a debate between sciences and culture]. PMID- 14752368 TI - [What about the ISAT results?]. PMID- 14752369 TI - [Commenting comments: to do or not to do?]. PMID- 14752370 TI - [Non-traumatic intracranial hemorrhage]. PMID- 14752371 TI - [Epidemiology of hemorrhagic strokes]. AB - Hemorrhagic stroke (HS) may be secondary to spontaneous intracerebral hemorrhage or to subarachnoid hemorrhage. There are no clinical symptoms that differentiate hemorrhagic stroke form ischemic stroke (IS) and only imaging can make the difference. But epidemiology, pathologic process and management are different in HS and IS. We will discuss the different risk factors of spontaneous intracerebral hemorrhage and subarachnoid hemorrhage. PMID- 14752372 TI - [Small vessel pathology and cerebral hemorrhage]. AB - Spontaneous intracerebral hemorrhage may be due to two main arteriolopathies: HTA and age-related small vessel disease and cerebral amyloid angiopathy. Historical study of the CHARCOT-BOUCHARD microaneurysm controversy leads to conclude that altered arterioles may be ruptured with or, probably more often, without microaneurysms. Recent neuroradiological studies with gradient-echo T2*W MR imaging have confirmed a classical neuropathological fact: there is a strong correlation between subcortical ischemic lesions (lacune, leukoaraiosis) and subcortical hemorrhage (hematomas, microbleeds). Sporadic Cerebral Amyloid Angiopathy is the first cause of spontaneous intracerebral hemorrhage after 70 years. Hemorrhages are lobar, typically recurrent and/or associated with cortical microbleeds at T2*W imaging. PMID- 14752373 TI - [Imaging of non-traumatic intracerebral hematoma]. AB - Intracerebral hematoma is mainly due to the spontaneous rupture of small vessels damaged by chronic hypertension or amyloid angiopathy. In some cases, intracerebral hemorrhage may be associated with a vascular malformation, a tumor, venous thrombosis or hemorrhagic transformation of a cerebral infarct. The objective of brain imaging is to identify the hematoma according to its different stages and to find a potential underlying cause because of the risk of recurrence and the possibilities of treatment. In emergency, a diagnosis of hematoma may be obtained by CT scan or MRI but the etiologic work-up requires early MRI. According to the patient's age, the medical history and the location of the hematoma, it may be necessary to perform conventional angiography in order to exclude an intracranial vascular malformation. The aim of this review is to detail the different aspects of intracerebral hemorrhages and to discuss the main causes that can be found at brain imaging. PMID- 14752374 TI - [Origin and consequences of the hypodensity surrounding spontaneous intracerebral hemorrhage in the acute phase]. AB - In the acute phase, spontaneous intracerebral hemorrhage is surrounded by hypodensity, the origin of which is controversial. Its clinical consequences are still under evaluation. Surrounding cerebral ischemia was suggested as its main etiology, but no experimental or clinical data currently supports this etiology. The hypodensity reflects the presence of cytotoxic and vasogenic edema secondary to prolonged contact between hemorrhage and adjacent brain tissue. This would mainly be due to local release of activated thrombin. However, other metabolites released from blood degradation products would also be involved. This edematous process is unique due to its early occurrence (as early as 2 hours), early maximum peak (24 hours), and regression over the next few days. In humans, no definitive proof exists that this edematous process of variable intensity is pathological. No specific treatment is currently available. PMID- 14752376 TI - [Prognosis and treatment of spontaneous intracerebral hematoma: review of the literature]. AB - The natural history of spontaneous intracerebral hematomas is difficult to precisely ascertain because of the heterogeneous nature of published series. However, the early evolution of these hematomas, within the first 24 hours, is now well known, especially with regards to predictive factors for early deterioration. Prognosic factors of longer-term evolution include alteration in the level of consciousness, as determined by the Glasgow score, the volume of the intracerebral hematoma, and the presence of intraventricular rupture. Volumetric data must be interpreted based on the location of the hematoma. Randomized studies about the management of intracerebral hematomas are rare. The beneficial value of early global management in a stroke unit (non-surgical) is undeniable and statistically proven. The value of some types of medical management (steroids, osmotherapy, preventive heparin therapy) was assessed by randomized trials with results that often were discordant. The main unknown treatment variable remains the role of surgical management. Eight randomized trials and following meta-analyses were not conclusive regarding the value of surgical management. The main difficulty does not relate to surgical technique but to the determination of clinical and radiological criteria for the selection of patients that are candidate to a surgical treatment. Available data will be reviewed. PMID- 14752378 TI - [Neurosurgical management of spontaneous cerebral hemorrhage]. AB - Intracerebral hemorrhage (ICH) accounts for 10 to 20% of strokes, but carries the highest rate of morbidity and mortality. Until now, there is no proven benefit in the literature for surgical treatment of ICH, and management of ICH varies greatly in neurosurgical centers. Surgery can be performed through standard craniotomy, or with a stereotactic procedure for deep-seated ICH. Conscious patients with minimal neurological deficit and small ICH are nonsurgical candidates. Patients with a Glasgow Coma Score lower than 4, with large deep seated ICH are also non surgical candidates. In other situations, the following arguments could lead to the decision of surgery: superficial (so-called lobar) ICH, size above 3 cm in diameter, midline shift, secondary neurological worsening, young patient, underlying vascular malformation. Acute hydrocephalus from ventricular hemorrhage may be treated with external ventricular drainage if the associated deep-seated ICH is small in size. Indications of surgery are more frequent for cerebellar ICH, as the risks for brainstem compression and hydrocephalus from ventricular obstruction are important. PMID- 14752379 TI - Unruptured intracranial aneurysms. AB - Between 3.6 and 6% of the population harbour an unruptured intracranial aneurysm. Risk of rupture is related to aneurysm site and size and whether or not the patient has already had a subarachnoid haemorrhage (SAH) from another aneurysm. In ISUIA 2, the rupture rate for anterior circulation aneurysms<7mm was 0% per year in patients with no prior SAH, and 0.3% per year in patients with previous SAH; 7-12mm aneurysms, 0.5% per year (both groups); 13-24mm aneurysms, 3% per year; and giant aneurysms 8% per year. Rupture rate for posterior circulation aneurysms is higher at all sizes:<7mm was 0.5% per year in subjects with no prior SAH, 0.7% in those with prior SAH; 7-12mm, 3% per year; 13-24mm, 3.7% per year; and giant aneurysms, 10% per year. Non-invasive tests like contrast enhanced magnetic resonance angiography (MRA) and multislice computed tomographic angiography (CTA) are alternatives to intra-arterial digital subtraction angiography (IADSA) to detect aneurysms. Although these are promising techniques, the quality of data testing their accuracy remains limited and single slice CTA and time-of-flight MRA are poorer at detecting aneurysms<5mm diameter, which account for up to 1/3 of unruptured aneurysms. For ruptured aneurysms, the only large scale randomised controlled trial comparing surgical and endovascular treatment (ISAT) by coiling, resulted in an absolute 8.8% reduction (updated figure as of June 2003 for 1888 patients) in death or dependency at 1 year compared with surgical clipping. For unruptured aneurysms, the best available data so far comparing coiling and clipping is from the prospective (but non randomised) arm of ISUIA. Elective surgical clipping had combined morbidity and mortality at 1 year of 12.2% versus 9.5% for coiling, although the groups were not matched with more high risk patients in the endovascular treatment cohort. Nevertheless these data are encouraging for future randomised trials of elective coiling versus clipping for asymptomatic aneurysms, in particular as the unproven long-term durability of coiling treatment and the fact that complete aneurysm occlusion is not always achieved remain obstacles to its wider use in unruptured aneurysms. There is an increased risk of SAH in relatives of patients with SAH (highest in those with two or more first degree relatives affected), but most SAH is sporadic and therefore the balance of available evidence indicates that mass screening for aneurysms is not cost effective. There may be a limited role for investigation of high-risk subgroups and ideally such screening should be tested in a randomised trial. The avoidance and active management of vascular risk factors should also be part of the management of at risk subjects. PMID- 14752380 TI - [Interview with Dr. Jean-Rene Hazard, Director of Public Relations, Guidant France]. PMID- 14752381 TI - [CD-ROM in pathology: another teaching tool]. PMID- 14752382 TI - [Quality assessment of intraoperative frozen sections in a teaching hospital: an analysis of 847 consecutive cases]. AB - We evaluated the accuracy of 847 consecutive frozen section diagnoses in order to develop a quality control. We also evaluated the time needed to perform them. Frozen sections and final diagnoses agreed in 92.6% and disagreed in 1.7% (14 cases). 5.8% of the cases were deferred. The only case of false-positive frozen sections (0.1%) was due to a pathologist's misinterpretation. False-negative frozen sections were due to sampling errors: in 5 cases, diagnostic tissue was present only in permanent sections of the frozen block and in 8 cases diagnostic tissue was present only in the portion of the specimen not sampled by the frozen section. One hundred and ninety two frozen sections concerned thyroid neoplasms. Thirteen cancers were diagnosed on frozen sections, 2 cancers were considered as benign and 9 cancers had a differed diagnosis. The mean duration to perform the frozen sections was 21 minutes. In conclusion, intra operative frozen section diagnosis is rapid and reliable. Discrepancies are more often false negatives due to sampling errors. Although a high rate of differed diagnosis was observed in thyroid neoplasms, frozen sections remain useful for these lesions. Imprint cytology of thyroid nodules is advisable. PMID- 14752383 TI - [Place of pathology in the forensic diagnosis of drowning]. AB - The diagnosis of drowning is one of the most difficult in forensic pathology. Drowning is death through the aspiration of fluid into the air passages. Signs of immersion only demonstrate submersion of the body for a period of time but are not signs of drowning. The best signs of drowning are froth around the mouth and nostrils and lung distension. Lung histology in drowning victims shows non specific lesions such as "emphysema aquosum" and alveolar edema. The diatom test for drowning requires a careful protocol to avoid possible contamination. Sample collection (cardiac blood, lung, liver, kidney tissue, bone marrow and brain) should be performed during the first part of the autopsy. Sample preparation requires chemical or enzymatic digestion. The diatom test should be both qualitative and quantitative. Species recovered from the body must correspond to those from the site of drowning and approximately in the same proportions. The diatom test has poor sensitivity and thus is much debated because it is always difficult to rule out possible contamination. Moreover, a negative diatom test cannot rule out drowning as the cause of death. The diagnosis of drowning is based on police investigations, forensic autopsy, microscopic analysis, and biochemical tests, but never solely on pathology findings. PMID- 14752384 TI - [The neuropathology of HIV infection in the era of highly active antiretroviral therapy]. AB - Introduction of Highly Active Antiretroviral Treatment (HAART) which is available for most AIDS patients in France since 1996, has resulted in a dramatic improvement of the disease course. From the survey of our autopsy series of (AIDS) cases and the review of other neuropathological studies from different developed countries, we found quantitative and qualitative changes in the pattern of human immunodeficiency virus (HIV) neuropathology. Quantitatively, there was a dramatic decrease in the number of autopsy cases but brain involvement remained a major cause of death in AIDS patients. There was an overall decrease of cerebral toxoplasmosis, cytomegalovirus encephalitis (CMVE) and HIV encephalitis (HIVE) for which successful treatment is available. This contrasted with the unchanged incidence of progressive multifocal leucoencephalopathy (PML) and primary malignant non Hodgkin brain lymphomas (PMBL). However, when looking closer at the last three years, the incidence of diseases affecting patients with severe immunodepression (CMVE, PML, PMBL) decreased in 2000-2002, whereas infections occurring in patients with milder immunodeficiency (toxoplasmosis, varicella zoster encephalitis (VZVE) or herpes simplex virus encephalitis (HSVE) became more frequent. Qualitatively, there were uncommon types of brain infections, such as BK virus encephalitis or general paresis. Finally, new forms of HIVE were reported: severe leukoencephalopathy with intense perivascular macrophage and lymphocyte infiltration possibly due to an exaggerated response from a newly reconstituted immune system; and also chronic "burnt out" forms of HIVE as VZVE, toxoplasmosis, or PML in which no inflammation and no infectious agent could be detected, likely due to prolonged survival. PMID- 14752385 TI - [Amphicrine carcinoma of the pancreas. Report of two cases]. AB - Amphicrine carcinomas are rare tumors defined by the presence of tumor cells showing evidence of both exocrine and endocrine differentiation. We here report two cases of amphicrine carcinomas of the pancreas, an exceedingly rare localization for this type of tumors. Diagnosis was made in respectively, a 32 year-old woman and a 66-year-old man; tumors measured 7 and 3 cm in diameter; metastatic dissemination was present in both cases. The first patient, treated by surgery and chemotherapy, is alive, without disease progression, after 26 months; the second patient deceased early after the diagnosis. In both cases, the first diagnosis considered at cytological and histological examination was endocrine carcinoma. The amphicrine nature of the lesion was ascertained by the combined demonstration of mucus staining and chromogranin A expression in the same cells. In one case, the amphicrine nature of tumor cells was confirmed by the ultrastructural examination. The identification of the amphicrine nature of an apparently endocrine tumor is of relevance, because of the poor prognosis of amphicrine carcinomas as compared to endocrine carcinomas and the requirement for aggressive therapy. PMID- 14752386 TI - [BK virus-induced tubulo-interstitial nephritis in a renal transplant recipient]. AB - We present a case of renal BK virus infection with renal allograft dysfunction. Renal allograft biopsy showed mononuclear infiltrates in the interstitium and viral inclusions in the tubular epithelial cells. Infected cells were stained with an anti-polyomavirus antibody. The polymerase chain reaction (PCR) performed on blood, urine, and on the DNA extracted from renal tissue showed the presence of the BK virus DNA sequence. The immunosuppressive therapy including tacrolimus, prednisone, and mycophenolate mofetil was reduced leading to an improvement of the renal function. BK virus infection is now recognized as a cause of renal allograft dysfunction, and has been observed with increasing frequency in recent years. Reactivation of the latent virus occurs in immunocompromised hosts such as organ recipients with immunosuppressive treatment. Histologically, renal BK virus infection is characterized by a lymphocytic interstitial infiltrate, and could mimic acute rejection. The pathologist should diagnose the viral infection and may be helped by urine cytology and immunohistochemistry. An accurate diagnosis is important because antirejection therapy favors the decline of the renal function. Enhanced new immunotherapy protocols seem to be the main risk factor for this infection. The response to reduced immunosuppression is variable with reports of an end stage renal failure in 70% of the patients after 18 months. PMID- 14752387 TI - [Pleomorphic carcinoma of the large intestine]. AB - Pleomorphic carcinoma, firstly described in the lung, is exceptional in the digestive tract. This tumor is often associated with fever and peripheral blood leukocytosis. The histological feature is characterized by giant pleomorphic often multinucleated cells with dense neutrophil infiltration and frequent phagocytosis of neutrophils by the tumor cells. Expression of epithelial markers confirm the diagnosis. The prognosis is very poor. We hereby report one case of pleomorphic carcinoma unusually situated in the large intestine, emphasizing difficulties encountered in establishing diagnosis. PMID- 14752388 TI - [Keratoacanthoma: two cases with intravascular spread]. AB - We report two patients with keratoacanthoma, simple in one and multiple in the other, displaying typical histological features except for intravascular spread. Although this spread points to malignancy, it did not allow to rule out the diagnosis of keratoacanthoma. These aggressive histological features, as well as perineural invasion, are not linked to malignant clinical course, according to the literature. Intravascular spread suggests that keratoancanthoma could be considered as a peculiar form of well differentiated squamous cell carcinoma. Comparison between clinical and pathological observations should lead to more specific diagnosis. PMID- 14752389 TI - [Invasive hydatiform mole in a postmenopausal woman]. AB - Gestational trophoblastic disease occurs rarely in postmenopausal women. We report the case of a 51 year-old postmenopausal woman with an invasive complete mole. Invasive mole should be distinguished from choriocarcinoma, by a thorough sampling showing infiltrative molar villi associated with a prominent trophoblastic proliferation. Gestational trophoblastic diseases in postmenopausal women can represent malignant changes of trophoblastic remnants of a prior pregnancy after a period of latency or correspond to a possible current pregnancy as demonstrated by an ovarian corpus luteum of pregnancy in our patient. The unusual finding in our case is that the gestational trophoblastic disease follows a pregnancy occurring after a biologically confirmed menopause. PMID- 14752391 TI - [A complicated ulcerative colitis]. PMID- 14752390 TI - [Epithelioid hemangioma of bone: a rare pathologic entity]. AB - Epithelioid hemangioma of bone is a rare benign vascular lesion. Since its first description by Rosai in 1979 about thirty cases have been reported. We report two new cases diagnosed in patients aged 21 and 7 years. Both patients had multiple lesions on the legs. Bone pain was the main symptom. Routine laboratories studies were unremarkable. Bon radiographs showed an expansive process of the bone. Histological and immunochemical features were typical of epithelioid hemangioma of the bone. Treatment consisted in above knee amputation for the first patient and therapeutic abstention for the second. Both patients are alive without progressive local disease or metastasis. PMID- 14752392 TI - [An ambiguous ovarian tumor]. PMID- 14752393 TI - [Gastrointestinal bleeding: a difficult diagnosis]. PMID- 14752394 TI - [A rare endothoracic tumor]. PMID- 14752395 TI - [2001 Standards, Options and Recommendations: practice guidelines for difficult diagnoses in surgical pathology or cytopathology in cancer patients]. AB - CONTEXT: The Standards, Options and Recommendations (SOR) project, which started in 1993, is a collaboration between the Federation of French Cancer Centers (FNCLCC), the 20 French Regional Cancer Centers, and specialists from French public universities, general hospitals and private clinics. The main objective is the development of clinical practice guidelines to improve the quality of health care and the outcome of cancer patients. OBJECTIVES: To elaborate practice guidelines for difficult diagnoses in surgical pathology or cytopathology in cancer patients. METHODS: The methodology is based on a literature review and critical appraisal by a multidisciplinary group of experts who define the CPGs according to the definitions of the Standards, Options and Recommendations project. Once the guidelines has been defined, the document is submitted for review by independent reviewers. RESULTS: The main recommendations to prevent and reduce the number of difficult diagnoses in surgical pathology or cytopathology are: The main recommendations to detect lesions associated with difficult diagnosis in surgical pathology or cytopathology are: The main recommendations to solve difficult diagnosis in surgical pathology or cytopathology are: PMID- 14752396 TI - [Intracapsular carcinoma ex mixed tumor]. PMID- 14752399 TI - [New perspectives in diagnosis and therapy of endocrine gastroenteropancreatic (GEP) tumors with somatostatin analogues]. AB - In the last decade important progresses have been obtained in the diagnosis and therapy of endocrine gastroenteropancreatic (GEP) tumors, mainly derived from the somatostatin receptors characterization and the introduction of long acting somatostatin analogues. Receptorial scintigraphy with radio-labeled analogues (Octreoscan) is the first choice investigation for staging and follow-up of endocrine GEP tumors, thanks to the high sensitivity in revealing the primary tumor and metastases, and for its capability to reveal lesions that are not identified by other imaging methods. Moreover, somatostatin analogues uptake by tumors allow us to use radiopharmaceutical compounds for advanced disease treatment. Between the radio-labeled drugs until now studied, interesting results have been obtained by DOTA-lanreotide (MAURITIUS), DOTA0 Tyr3-octreotide (DOTATOC) and DOTA0 Tyr3-octreotate, bound to beta-emitting radio-isotope suitable for therapeutic use. In the field of the pharmacological therapy of GEP tumors, the clinical trials show that somatostatin analogues reduce the symptoms related to functionally active tumors and stabilize or slow tumor growth improving the patient quality of life. Although somatostatin analogues alone could not be able to cure GEP tumors, their early utilization in association with surgical debulking of primary tumor and metastases, embolization or chemoembolization, and interferon, chemotherapy and radio-metabolic therapy (mainly directed to the destruction of micrometastases), increases the possibility of a radical therapeutic intervention. The continuous evolution of pharmacological research provides always new analogues (octreotide LAR, lanreotide, vapreotide, BIM-23244, BN 81644, PTR-3173, BIM-23A387, SOM-230, etc.) with different pharmacokinetic and receptorial properties and acting with more effectiveness in the different individual clinical situations. In this context there have been recently introduced also the "chimeric" analogues. On the other hand, the widespread utilization of molecular biology and immunohistochemical methods can allow, in perspective, to better define the receptorial pattern of individual endocrine tumors, after their surgical removal. The necessity to integrate endocrinological, nuclear medicine, surgical, oncologic and laboratory competencies behaves a multidisciplinary approach based on the utilization of diagnostic-therapeutic protocols supplying comparable results. It does not appear unjustified to expect, in the future, a scenery of more "individual" and more effective therapies for patients affected by GEP tumours. PMID- 14752400 TI - [Immunologic and genetic aspects of latent autoimmune diabetes in the adult]. AB - The presence of islet cell autoantibodies (ICA), and especially of glutamic acid decarboxylase autoantibodies (GAD65Ab), in patients with non-insulin-dependent diabetes mellitus identifies the so-called latent autoimmune diabetes in the adult (LADA). LADA patients have an increased risk for developing insulin deficiency, and in 60-80% of cases the exogenous insulin therapy must be started within 5-6 years. GAD65Ab identify a subgroup of type 2 diabetic (T2DM) patients with low body mass index (BMI) at the time of diagnosis. The presence of GAD65Ab at high titres and directed against COOH-terminal epitopes of the autoantigen, or the presence of both GAD65Ab and ICA, discriminates patients with clinical characteristics very similar to those of a slowly progressive form of type 1 diabetes (T1DM). On the other hand, the presence of low levels GAD65Ab, in the absence of ICA or other immune markers, such as IA-2 antibodies, characterizes a subgroup of patients with clinical characteristics almost indistinguishable from those of typical T2DM patients. The autoimmune origin of LADA is also demonstrated by the increased frequency of thyroid and adrenal autoantibodies, as compared to GAD65Ab-negative T2DM patients, and by the strong genetic association with HLA-DR3-DQ2, -DR4-DQ8 and the polymorphisms of the MHC class I chain-related A (MICA) and CTLA-4 genes. Metabolic studies have shown the coexistence of insulin resistance and insulin secretion defect supporting the hypothesis that LADA may be the result of the interaction of a genetic background predisposing for islet autoimmunity and a genetic background predisposing for T2DM. PMID- 14752401 TI - Melatonin and energy homeostasis: peripheral versus central regulation. AB - Melatonin, the hormone of darkness, has been known for a long time to be a major regulator of energy homeostasis in hibernating animals. Much less is known about the role of melatonin in energy homeostasis in non-hibernating animals, including humans. In mammals, two specific melatonin receptor subtypes, MT1 and MT2, have been cloned and are known to be expressed at central and peripheral sites. Although a central regulation of energy homeostasis has been widely accepted for hibernating animals, the exact site of melatonin action remains still poorly defined. Central effects appear to be predominantly mediated by the MT1 subtype. Recently, several groups showed that melatonin may also have a direct effect on peripheral tissues involved in energy homeostasis such as pancreatic beta cell, hepatocytes and adipocytes. Both, the MT1 and MT2 subtypes appear to be involved. The respective contribution of central and peripheral effects of melatonin on energy homeostasis in vivo must be established in future studies. PMID- 14752402 TI - Ectopic uterine stem cell tumors in the hamster kidney. A unique model for estrogen-induced oncogenesis. AB - Estrogens are intimately involved in the causation of some of the most prevalent cancers afflicting women, particularly, breast, endometrial, cervico-vaginal, and possibly ovarian. Therefore, it has become particularly pertinent to elucidate the molecular mechanisms whereby estrogens elicit their oncogenic actions so that better prevention strategies can be developed. The estrogen-induced Syrian hamster tumors of the kidney have emerged as one of the most intensively studied in vivo models in solely estrogen oncogenesis. An advantage of this model is that the tumors occur in the absence of any intervening morphologic changes, but rather they are the result of the continuous progression of some interstitial stem cells in the kidney leading to tumor formation. Evidence is presented that the origin of these tumors is derived from ectopic "uterine" stem cells, which are responsive to estrogenic hormones. Their steroid receptor and many other gene alterations have been delineated. Importantly, a crucial early event in this solely estrogen-induced oncogenic process is the overexpression and amplification of c-myc and its protein product. Chromosomal instability, in both early and large frank tumors, is another important characteristic of this process. This later feature has commonly been shown in solely estrogen-induced murine mammary tumors, and in ductal carcinomas in situ and in primary invasive ductal breast carcinomas. These changes are considered crucial in eliciting estrogen-induced tumorigenesis. PMID- 14752403 TI - Gender and valvular surgery. PMID- 14752404 TI - Valvular heart disease in women: the surgical perspective. PMID- 14752405 TI - Off-pump surgery and cerebral injury. PMID- 14752406 TI - Idiopathic laryngotracheal stenosis. PMID- 14752407 TI - The influence of pH strategy on cerebral and collateral circulation during hypothermic cardiopulmonary bypass in cyanotic patients with heart disease: results of a randomized trial and real-time monitoring. AB - OBJECTIVE: The optimal pH strategy during hypothermic cardiopulmonary bypass remains controversial. Systemic pulmonary collateral circulation may develop in patients with cyanotic anomalies. The purpose of this study was to evaluate the effect of pH strategies on cerebral oxygenation and systemic pulmonary collateral circulation during hypothermic cardiopulmonary bypass in cyanotic patients with heart disease. METHODS: Forty cyanotic patients (age > 1 year) with heart disease were prospectively randomized into 2 groups. Group 1 (n = 19, 14.3 +/- 1.5 kg) underwent hypothermic cardiopulmonary bypass with alpha-stat strategy and group 2 (n = 21, 12.5 +/- 0.9 kg) with pH-stat. Cardiopulmonary bypass was established with pump-assisted drainage. Cerebral oxygenation was assessed by near-infrared spectroscopy and the systemic pulmonary collateral circulation was calculated by pump flows [% systemic pulmonary collateral circulation = perfusion flow - drainage flow)/perfusion flow x 100]. Lactate was measured as an index of systemic anaerobic metabolism. RESULTS: There were no significant differences in preoperative hematocrit, oxygen saturation, Qp/Qs, cardiopulmonary bypass duration, minimum temperatures, perfusion flow and pressure, urine output, and depth of anesthesia between the groups. Oxyhemoglobin signal and tissue oxygenation index of near-infrared spectroscopy monitoring were significantly lower in group 1 compared with group 2 (P =.008 and P <.0001, respectively), suggesting inadequate cerebral oxygenation with alpha-stat. Deoxygenated hemoglobin signal was significantly higher in group 1 relative to group 2 (P <.0001). The % systemic pulmonary collateral circulation was significantly lower in group 2 compared with group 1, suggesting a reduced pulmonary collateral circulation with pH-stat (P <.0001, average; group 1, 20.1% +/- 1.2%; group 2; 7.7% +/- 0.7%). Serum lactate was significantly lower in group 2 (P <.0001). CONCLUSIONS: The pH-stat strategy results in an improved environment, including sufficient cerebral oxygenation, decreased systemic pulmonary collateral circulation, and lower lactate level during hypothermic cardiopulmonary bypass in cyanotic patients with heart disease. Future studies should investigate the long term neurological outcome. PMID- 14752408 TI - Nitric oxide synthase gene transfer inhibits biological features of bypass graft disease in the human saphenous vein. AB - BACKGROUND: Bypass graft disease is related to proliferation and migration of vascular smooth muscle cells and to platelet activation with thrombus formation. Nitric oxide inhibits these biological responses; it has never been demonstrated, however, whether this occurs in intact human vascular tissue after endothelial nitric oxide synthase gene transfer. METHODS: We examined whether endothelial nitric oxide synthase overexpression inhibits biological features of bypass graft disease in saphenous vein tissue. RESULTS: The nitric oxide donor diethylenetriamineNONOate inhibited proliferation (P <.001) and migration (P <.001) of human saphenous vein vascular smooth muscle cells in response to 20% serum in a concentration-dependent manner. A similar effect on proliferation (P <.05) and migration (P <.05) without any cytotoxicity was observed after adenoviral endothelial nitric oxide synthase transfection. Staining of saphenous vein tissue for placental alkaline phosphatase demonstrated that adenoviral transfection was efficient. Consistent with this observation, endothelial nitric oxide synthase protein expression and nitric oxide release were enhanced in transfected tissue. Further, endothelial nitric oxide synthase overexpression inhibited vascular smooth muscle cell outgrowth from saphenous vein explants over 21 days; 48% +/- 12% of explants exhibited outgrowth after treatment with endothelial nitric oxide synthase adenovirus as compared with 69% +/- 10% in those infected with control adenovirus and 90% +/- 5% in uninfected tissue (P <.05). Similarly, platelet adhesion to human saphenous vein tissue was inhibited by endothelial nitric oxide synthase overexpression; adhesion was reduced in segments infected with endothelial nitric oxide synthase adenovirus (58% +/- 6%) as compared with those infected with control adenovirus (107% +/- 8%) or uninfected saphenous vein (100%; P <.05). CONCLUSIONS: These data demonstrate that endothelial nitric oxide synthase gene transfer inhibits biological features of bypass graft disease in intact human saphenous vein tissue. Therefore, endothelial nitric oxide synthase transfection represents a promising gene transfer approach to prevent venous bypass graft disease. PMID- 14752409 TI - Gene delivery to aortocoronary saphenous vein grafts in a large animal model of intimal hyperplasia. AB - OBJECTIVE: More than 50% of aortocoronary saphenous vein grafts are occluded 10 years after surgery. Intimal hyperplasia is an initial, critical step in the progression toward occlusion. To date, no clinically relevant large animal models of aortocoronary saphenous vein graft intimal hyperplasia have been fully characterized. Gene therapy holds promise as a novel treatment for aortocoronary saphenous vein graft intimal hyperplasia. The 2 objectives of this study are to characterize a canine model of aortocoronary saphenous vein graft intimal hyperplasia and to demonstrate that ex vivo gene delivery is possible in these grafts using adenoviral vectors. METHODS: Ten dogs underwent aortocoronary bypass grafting using saphenous veins. Six dogs underwent serial arteriograms to monitor graft patency. On postoperative day 90, the dogs were killed and their grafted and nongrafted saphenous veins were studied histologically. Four dogs underwent the same procedure, but their saphenous veins were treated with 1 x 10(12) total viral particles of a replication-deficient, recombinant adenovirus containing beta-galactosidase (n = 2) or the beta-adrenergic receptor kinase carboxyl terminus (n = 2). These animals were killed on postoperative day 7 for determination of transgene expression. RESULTS: All grafts were demonstrated patent by arteriogram before the animals were killed. The mean intimal area of the saphenous vein grafts was increased when compared with that of the nongrafted saphenous veins (2.83 mm(2) vs 0.09 mm(2), P <.0008). Adenoviral-treated saphenous vein grafts demonstrated positive transgene expression either by X-gal staining (beta-galactosidase) or Northern analysis (beta-adrenergic receptor kinase carboxyl terminus). CONCLUSION: This study characterizes a clinically relevant canine model of aortocoronary saphenous vein graft intimal hyperplasia. In addition, it demonstrates that adenoviral vectors can be delivered ex vivo to the saphenous vein graft vessel wall at subphysiologic distension pressures. This model may be used in future studies to manipulate molecular targets critical in aortocoronary saphenous vein graft intimal hyperplasia. PMID- 14752410 TI - Intramyocardial and intracoronary basic fibroblast growth factor in porcine hibernating myocardium: a comparative study. AB - OBJECTIVE: Therapeutic angiogenesis is an alternative method of revascularization for end-stage coronary artery disease. We determined the effects of intramyocardial and intracoronary basic fibroblast growth factor 2 on myocardial blood flow and function in a porcine model of hibernating myocardium. METHODS: Twenty-four mini-swine with 90% left circumflex artery stenosis and documented hibernating myocardium by positron emission tomography and dobutamine stress echocardiography were randomized to intramyocardial basic fibroblast growth factor 2 at 0.6 microg/kg (mid-dose, n = 6, 30 injections/animal), 6 microg/kg (high-dose, n = 6, 30 injections/animal), or intramyocardial vehicle control (n = 6). The intracoronary group received 6 microg/kg basic fibroblast growth factor 2 (n = 6) into the right and left circumflex artery coronary arteries. Positron emission tomography and dobutamine stress echocardiography were repeated at 1 and 3 months. RESULTS: In the vehicle group, normalized left circumflex artery myocardial blood flow was 0.74 +/- 0.04 at 1 month and 0.75 +/- 0.07 at 3 months compared with 0.68 +/- 0.03 at baseline. In the intracoronary group, myocardial blood flow was 0.71 +/- 0.03 at 1 month and 0.72 +/- 0.04 at 3 months compared with 0.67 +/- 0.04 at baseline. In the mid group, myocardial blood flow was 0.73 +/- 0.06 at 1 month and 0.85 +/- 0.05 at 3 months (P <.001) compared with 0.67 +/ 0.04 at baseline. In the high group, myocardial blood flow was 0.81 +/- 0.06 at 1 month and 0.83 +/-.04 at 3 months (P =.03) compared with 0.71 +/- 0.02 at baseline. No significant improvements in ischemia were demonstrated in any of the groups by dobutamine stress echocardiography at 1 or 3 months. CONCLUSIONS: In porcine hibernating myocardium, intramyocardial basic fibroblast growth factor 2 significantly improved regional myocardial blood flow 3 months after treatment. There was no significant change in function in any of the 4 groups. These data suggest that intramyocardial dosing of basic fibroblast growth factor 2 (0.6 microg/kg) may be an optimal dose for improving perfusion in the treatment of end stage coronary artery disease. PMID- 14752411 TI - Supplemental nitric oxide and its effect on myocardial injury and function in patients undergoing cardiac surgery with extracorporeal circulation. AB - BACKGROUND: Cardiopulmonary bypass induces a systemic inflammatory response that may contribute to clinical morbidity. Gaseous nitric oxide at relatively low concentrations may elicit peripheral anti-inflammatory effects in addition to a reduction of pulmonary resistances. We examined the effects of 20 ppm of inhaled nitric oxide administered for 8 hours during and after cardiopulmonary bypass. METHODS AND RESULTS: Twenty-nine consecutive patients undergoing aortic valve replacement combined with aortocoronary bypass were randomly allocated to either 20 ppm of inhaled nitric oxide (n = 14) or no additional inhalatory treatment (n = 15). Blood samples for total creatine kinase, creatine kinase MB fraction, and troponin I measurements were collected at 4, 12, 24, and 48 hours postsurgery. In addition, we collected perioperative blood samples for measurements of circulating nitric oxide by-products and brain natriuretic peptide. Soluble P selectin was analyzed in blood samples withdrawn from the coronary sinus before and after aortic clamping. The area under the curve of creatine kinase MB fraction (P =.03), total creatine kinase (P =.04), and troponin I (P =.04) levels were significantly decreased in the nitric oxide-treated patients. Moreover, in the same group we observed blunted P-selectin and brain natriuretic peptide release (P =.01 and P =.02, respectively). Nitric oxide inhalation consistently enhanced nitric oxide metabolite levels (P =.01). CONCLUSIONS: Nitric oxide, when administered as a gas at low concentration, is able to blunt the release of markers of myocardial injury and to antagonize the left ventricular subclinical dysfunction during and immediately after cardiopulmonary bypass. The organ protection could be mediated, at least in part, by its anti-inflammatory properties. PMID- 14752412 TI - Improved cerebral protection through replacement of residual intracavital air by carbon dioxide: a porcine model using diffusion-weighted magnetic resonance imaging. AB - BACKGROUND: Major risk of central or peripheral organ damage is attributed to air embolism from incompletely de-aired cardiac chambers after cardiac operations. Replacement of air by carbon dioxide insufflation into the thoracic cavity is widely used. Diffusion-weighted magnetic resonance imaging of the brain detects ischemia within minutes after onset. The reversibility of ischemia in cerebral tissue after massive gaseous emboli has not yet been described. METHODS: After selective catheterization of a common carotid artery in 15 pigs, boli of 1 mL/kg body weight of air (n = 5) or carbon dioxide (n = 5, "low dose") were applied. Five pigs received 2 mL/kg body weight of carbon dioxide ("high dose"). Diffusion weighted magnetic resonance imaging of the brain was performed 2, 5, 10, 15, and 25 minutes after embolization. RESULTS: All animals of the "air" group showed important circulatory reactions leading to death of 2 animals. In the whole group, diffusion-weighted magnetic resonance imaging revealed irreversible hyperintense signals in both hemispheres. In the low-dose group, no change in signal intensity was observed in 2 pigs, and 3 others showed reversible changes in signal intensity, without important circulatory reactions. In 3 animals of the high-dose group, hyperintense signals were reversible, but 2 others presented with bilateral, irreversible signals in diffusion-weighted magnetic resonance imaging, accompanied by minor circulatory reactions. CONCLUSION: In contrast to the dramatic effect of air emboli, identical quantities of carbon dioxide injected into cerebral arteries of the pigs were not associated with major clinical symptoms. The early reversibility of ischemic reactions visualized in diffusion-weighted magnetic resonance imaging encourages the use of carbon dioxide insufflation as a protective method in cardiac surgery. PMID- 14752413 TI - Predictors of delirium after cardiac surgery delirium: effect of beating-heart (off-pump) surgery. AB - BACKGROUND: Despite improved outcomes after cardiac operations, postoperative delirium remains a common complication that is associated with increased morbidity and prolonged hospital stay. METHODS: Univariate and multivariate predictors of postoperative delirium were determined from prospectively gathered data on 16,184 patients undergoing cardiac operations with cardiopulmonary bypass (conventional, n = 14,342) and without cardiopulmonary bypass (beating-heart surgery, n = 1847) between April 1996 and August 2001. Delirium was defined as a transient mental syndrome of acute onset characterized by global impairment of cognitive functions, a reduced level of consciousness, attentional abnormalities, increased or decreased psychomotor activity, and a disordered sleep-wake cycle. RESULTS: The overall prevalence of postoperative delirium was 8.4%. Of 49 selected patient-related risk factors and treatment variables, 35 were highly associated with postoperative delirium by univariate analysis. Stepwise logistic regression revealed the following variables as independent predictors of delirium: history of cerebrovascular disease, peripheral vascular disease, atrial fibrillation, diabetes mellitus, left ventricular ejection fraction of 30% or less, preoperative cardiogenic shock, urgent operation, intraoperative hemofiltration, operation time of 3 hours or more, and a high perioperative transfusion requirement. Two variables were identified as having a significant protective effect against postoperative delirium: beating-heart surgery and younger patient age. CONCLUSIONS: Postoperative delirium is a common complication in cardiac operations. The increased use of beating-heart surgery without cardiopulmonary bypass may lead to a lower prevalence of this complication and thus improve patient outcomes. PMID- 14752414 TI - Traction injury during minimally invasive harvesting of the saphenous vein is associated with impaired endothelial function. AB - OBJECTIVE: Many methods of minimally invasive surgical harvesting of the great saphenous vein have been developed because of the morbidity related to the long skin incision after traditional (open) great saphenous vein harvesting. One such method involves the use of multiple small incisions separated by 10- to 15-cm skin bridges through which the saphenous vein is harvested. We hypothesized that this method of saphenous vein harvesting might subject the saphenous vein to considerable traction forces, resulting in impaired endothelial cell function. METHODS: Four-millimeter great saphenous vein segments were obtained from patients undergoing elective coronary artery bypass graft surgery. Group A (minimally invasive surgery) consisted of 23 rings from 20 patients (age, 65.8 +/ 11.1 years, mean +/- SD). Group B (open harvesting) consisted of 33 rings from 8 patients (age, 69.8 +/- 8.6 years). All great saphenous vein segments were undistended and were used within 24 hours of harvesting. Isometric tension experiments were performed on each ring of the great saphenous vein by using a force-displacement transducer to measure the force of contraction in grams. Measurements included developed force after exposure to high-potassium depolarizing solution and 50 micromol/L phenylephrine and decrease in force of contraction (relaxation) after exposure to 1 and 10 micromol/L acetylcholine. RESULTS: There were no differences between the minimally invasive surgery and open harvesting groups in their responses to high-potassium depolarizing solution or phenylephrine: high-potassium depolarizing solution, contractions of 4.26 +/- 0.72 g (mean +/- SEM) and 3.95 +/- 0.38 g, respectively (P =.70); phenylephrine, contractions of 3.49 +/- 0.63 g and 2.73 +/- 0.39 g, respectively (P =.41). There was no net relaxation in segments from the minimally invasive surgery group after exposure to 1.0 or 10 micromol/L acetylcholine. In contrast, rings from the open harvesting group demonstrated relaxation of -0.41 +/- 0.07 g and -0.32 +/- 0.09 g after exposure to 1.0 and 10 micromol/L acetylcholine, respectively. CONCLUSIONS: In undistended saphenous vein segments isolated from patients undergoing minimally invasive surgical and open techniques of harvesting, there was no acetylcholine-mediated endothelium-dependent relaxation in the minimally invasive surgery group. Therefore harvesting of the great saphenous vein through multiple small incisions might result in endothelial dysfunction, possibly caused by traction injury. PMID- 14752415 TI - Attenuation of adhesion formation after cardiac surgery with a chymase inhibitor in a hamster model. AB - OBJECTIVE: Chymase is one of the inflammatory mediators and is released from mast cells, which are closely associated with adhesion formation. Chymase also activates transforming growth factor beta1, which promotes tissue fibrosis. However, the role of chymase in cardiac adhesion formation has not yet been elucidated. We have assessed whether a specific chymase inhibitor, Suc-Val-Pro Phe(p) (OPh)(2), prevents postoperative cardiac adhesions in hamsters. METHODS: In 66 hamsters the epicardium was abraded, and then either chymase inhibitor or placebo was injected into the left thoracic cavity, leaving the pericardium open. Cardiac chymase activity, the level of transforming growth factor beta1 in the pleural fluid, and the density of epicardial mast cells were measured 3 days postoperatively. The degree of adhesion formation was evaluated macroscopically and histologically 2 weeks postoperatively by using a grading score ranging from 0 (no adhesions) to 4 (severe adhesions). RESULTS: The cardiac chymase activity and level of transforming growth factor beta1 were lower in the chymase inhibitor treated group compared with in the placebo-treated group (45.8 +/- 18.7 vs 79.7 +/- 13.7 microU/mg protein [P <.025] and 15.6 +/- 6.5 vs 33.2 +/- 9.8 microg/mL [P <.01], respectively). The density of mast cells was higher in the placebo treated group, and there was suppression to 60% of this value in the chymase inhibitor-treated group. The adhesion scores were lower in the chymase inhibitor treated group compared with in the placebo-treated group (1.3 +/- 1.3 vs 3.0 +/- 1.1, P <.01). CONCLUSION: Use of a chymase inhibitor suppresses not only cardiac chymase activity but also the level of transforming growth factor beta1, and this results in a reduction in postoperative cardiac adhesion. PMID- 14752416 TI - A phase II dose-response study of hemoglobin raffimer (Hemolink) in elective coronary artery bypass surgery. AB - BACKGROUND: We performed this study to determine the dose-response of hemoglobin raffimer administered in conjunction with intraoperative autologous donation in patients undergoing coronary artery bypass grafting surgery. A secondary objective was to evaluate hemoglobin raffimer for reducing the incidence of allogeneic red blood cell transfusions. METHODS: This was a phase II, single blind, multicenter, placebo-controlled, open-label study. Patients undergoing coronary artery bypass grafting with cardiopulmonary bypass and intraoperative autologous donation were randomized to receive a single dose of hemoglobin raffimer or control (10% pentastarch). Patients were sequentially enrolled in a dose block of 250, 500, 750, and 1000 mL. RESULTS: Sixty patients received hemoglobin raffimer (n = 30) or control (n = 30). Hemoglobin raffimer was well tolerated. Most (98%) adverse events were mild or moderate in severity. There was an expected dose-dependent increase in the incidence of blood pressure increases and jaundice in hemoglobin raffimer-treated patients. In a dose-pooled analysis of hemoglobin raffimer versus control, increased blood pressure (43% vs 17%), nausea (37% vs 33%), and atrial fibrillation (37% vs 17%) were the most frequently reported adverse events. All serious adverse events were considered unrelated or unlikely to be related to study drug. No hemoglobin raffimer-treated patient required an intraoperative allogeneic red blood cell transfusion, compared with 5 (17%) pentastarch-treated patients (P =.052). This advantage of hemoglobin raffimer was maintained at 24 hours after surgery (7% vs 37%; P =.010) and up to 5 days after surgery (10% vs 47%; P =.0034). CONCLUSIONS: Hemoglobin raffimer was not associated with any serious adverse events in patients undergoing primary coronary artery bypass grafting with cardiopulmonary bypass and intraoperative autologous donation in a dose-response study up to 1000 mL. Hemoglobin raffimer was effective in facilitating decreased exposure or avoidance of allogeneic red blood cell transfusions when used in conjunction with intraoperative autologous donation. PMID- 14752417 TI - Comparison of mutational changes in involved N1 lymph nodes with those in primary tumors in stage II non-small cell lung cancer: a pilot study. AB - OBJECTIVES: Surgical resection is the standard treatment for stage II non-small cell lung cancer, but recurrence rates approach 60%. This study compared mutational changes in involved lymph nodes and primary tumors from patients with stage II non-small cell lung cancer to determine whether risk factors for recurrence could be identified. METHODS: Forty patients with resected stage II non-small cell lung cancer (excluding T3 N0 disease) were studied. Microdissection was performed on primary tumors and lymph nodes. Analysis was performed across 9 genomic loci by using polymerase chain reaction amplification. The ratio of fractional allelic loss between involved lymph nodes and primary tumors was used to stratify patients into high-risk (fractional allelic loss ratio of >or=1) and low-risk (fractional allelic loss ratio of <1) groups. RESULTS: The median age of the patients was 68 years (range, 42-85 years). Median follow-up was 30 months. Fractional allelic loss was greater in patients with squamous carcinomas compared with that in adenocarcinomas, but survival was similar (35 vs 39 months). The median survival was 35 months in high-risk patients and was not reached in low-risk patients (P =.3). Disease-free survival was 24 months in high-risk patients and was not reached in low-risk patients (P =.35). In the subset with adenocarcinoma (n = 18), median survival was 24 months in the high-risk group; no deaths occurred in low-risk patients (P =.01). Also, disease-free survival was 14 months in high-risk patients and was not reached in the low-risk patients (P =.05). CONCLUSIONS: Squamous cancers demonstrate greater mutational changes than adenocarcinomas; this does not affect outcome. The patients with low-risk adenocarcinomas demonstrated superior outcomes compared with those of other patients. These results should be confirmed in larger studies. PMID- 14752418 TI - Down-regulation of beta catenin inhibits the growth of esophageal carcinoma cells. AB - INTRODUCTION: Esophageal cancer remains a highly lethal malignancy, with therapeutic options of limited efficacy in the majority of patients. Understanding the molecular events involved in the pathogenesis of esophageal cancer offers insight into potential targets for treatment. Beta catenin and Wnt signaling abnormalities are involved in the development of both adenocarcinoma and squamous carcinoma of the esophagus. We hypothesized that down-regulation of beta catenin would inhibit the growth of human esophageal cancer. METHODS: A human esophageal squamous cell carcinoma cell line (TE10) was treated with phosphorothioate antisense oligonucleotides to beta catenin. The cells were subsequently assayed for beta catenin mRNA and protein by real-time polymerase chain reaction and Western blot. Beta catenin transcriptional activity was determined by TOPFlash assay. Cell viability and growth was assessed by methyl thiazol-diphenyl-tetrazolium assay and trypan blue exclusion. A colorimetric assay was employed to assess caspase 3 activity, and flow cytometry was done to determine percentage of cells in a given phase of the cell cycle. RESULTS: Following antisense treatment, beta catenin mRNA and protein concentration were decreased. There was corresponding decrease in beta catenin-transcription factor dependent transcription. Treatment with beta catenin antisense resulted in significantly decreased cell viability and proliferation. The mechanism appears to be increased induction of apoptosis. CONCLUSIONS: These data suggest a potential role for the targeting of beta catenin in the treatment of esophageal cancer. PMID- 14752419 TI - Idiopathic laryngotracheal stenosis: effective definitive treatment with laryngotracheal resection. AB - OBJECTIVE: Little was known about idiopathic laryngotracheal stenosis when it was first described. We have operated on 73 patients with idiopathic laryngotracheal stenosis, have confirmed its mode of presentation and response to surgical therapy, and have established long-term follow-up. METHODS: Charts of 73 patients treated surgically for idiopathic laryngotracheal stenosis between 1971 and 2002 were retrospectively reviewed. RESULTS: All patients were treated with a single staged laryngotracheal resection, with (36/73) and without (37/73) a posterior membranous tracheal wall flap. Nearly all were women (71/73), with a mean age of 46 years (range, 13-74 years). Twenty-eight (38%) of 73 had undergone a previous procedure with laser, dilation, tracheostomy, T-tube, or laryngotracheal operations. After laryngotracheal resection, the majority of patients (67/73) were extubated in the operating room, and 7 required temporary tracheostomies, only 1 of whom was among the last 30 patients. All were successfully decannulated. There was no perioperative mortality. Principal morbidity was alteration of voice quality, which was mild and tended to improve with time. Sixty-seven (91%) of 73 patients had good to excellent long-term results with voice and breathing quality and do not require further intervention for their idiopathic laryngotracheal stenosis. CONCLUSION: Idiopathic laryngotracheal stenosis is an entity that occurs almost exclusively in women and is without a known cause. It is not a progressive process, but the timing of the operation is crucial. Single-staged laryngotracheal resection is successful in restoring the airway while preserving voice quality in more than 90% of patients. Protective tracheostomy is now rarely required (1/30). Long-term follow-up shows a stable airway and improvement in voice quality. PMID- 14752420 TI - Two commonly used neoadjuvant chemoradiotherapy regimens for locally advanced stage III non-small cell lung carcinoma: long-term results and associations with pathologic response. AB - BACKGROUND: We performed this study to determine the outcomes (pathologic response, survival, local-regional control, and toxicity) in patients treated with neoadjuvant chemoradiotherapy and planned operation for stage IIIA non-small cell lung carcinoma. METHODS: Patients treated from 1993 to 2000 with neoadjuvant chemoradiotherapy and a predetermined plan for subsequent surgical resection for stage III non-small cell lung carcinoma were analyzed. All patients underwent pretreatment evaluation at the university's Multidisciplinary Lung Cancer Center. Most patients (87%) had complete mediastinoscopy staging, and all were believed to be poor candidates for up-front operation because of bulky extent of disease. The radiotherapy program used conventional, 2-dimensionally planned treatment to 45 to 54 Gy in 1.8- to 2-Gy fraction size. Concurrent chemotherapy consisted of etoposide/cisplatin or carboplatin/paclitaxel. Study end points included resectability, pathologic response, local-regional control, survival, and toxicity. An exploratory comparison between pathologic response and long-term survival was performed. An exploratory comparison between older chemotherapy (etoposide/cisplatin) and third-generation chemotherapy (carboplatin/paclitaxel) was also performed. RESULTS: Of 53 patients, 45 (85%) were deemed surgical candidates after induction therapy. Twenty-two (42% of the initial cohort) patients had a major pathologic response to stage 0, I, or II disease. The 5-year actuarial survival was 31%. Major pathologic response was associated with improved survival (48% vs 24%; P =.027). The overall rate of early death potentially related to therapy in this series was 9%; this mostly occurred in patients who underwent right pneumonectomy. There was no difference in efficacy or mortality between etoposide/cisplatin and radiotherapy versus carboplatin/paclitaxel and radiotherapy, although the latter regimen was associated with less grade 3 or higher acute toxicity necessitating interruption or hospitalization during neoadjuvant treatment (P =.02). In-field local control was achieved in 83% of all patients (90% of the patients who underwent resection). Brain metastases as the first site of treatment failure occurred in 23% of all patients. CONCLUSIONS: Neoadjuvant concurrent chemoradiation delivers high resectability, major pathologic response rate, and excellent local-regional control, with encouraging long-term survival considering the patient population studied. Major pathologic response correlates with long-term survival. Neoadjuvant carboplatin/paclitaxel and radiotherapy is an appropriate framework on which to add new therapies. PMID- 14752421 TI - A decade of living lobar lung transplantation: recipient outcomes. AB - OBJECTIVE: Living lobar lung transplantation was developed as a procedure for patients considered too ill to await cadaveric transplantation. METHODS: One hundred twenty-eight living lobar lung transplantations were performed in 123 patients between 1993 and 2003. Eighty-four patients were adults (age, 27 +/- 7.7 years), and 39 were pediatric patients (age, 13.9 +/- 2.9 years). RESULTS: The primary indication for transplantation was cystic fibrosis (84%). At the time of transplantation, 67.5% of patients were hospitalized, and 17.9% were intubated. One-, 3-, and 5-year actuarial survival among living lobar recipients was 70%, 54%, and 45%, respectively. There was no difference in actuarial survival between adult and pediatric living lobar recipients (P =.65). There were 63 deaths among living lobar recipients, with infection being the predominant cause (53.4%), followed by obliterative bronchiolitis (12.7%) and primary graft dysfunction (7.9%). The overall incidence of acute rejection was 0.8 episodes per patient. Seventy-eight percent of rejection episodes were unilateral. Age, sex, indication, donor relationship, preoperative hospitalization status, use of preoperative steroids, and HLA-A, HLA-B, and HLA-DR typing did not influence survival. However, patients on ventilators preoperatively had significantly worse outcomes (odds ratio, 3.06, P =.03; Kaplan-Meier P =.002), and those undergoing retransplantation had an increased risk of death (odds ratio, 2.50). CONCLUSION: These results support the continued use of living lobar lung transplantation in patients deemed unable to await a cadaveric transplantation. We consider patients undergoing retransplantations and intubated patients to be at significantly high risk because of the poor outcomes in these populations. PMID- 14752422 TI - Immuno-gene therapy with interferon-beta before surgical debulking delays recurrence and improves survival in a murine model of malignant mesothelioma. AB - OBJECTIVES: Immuno-gene therapy of mesothelioma with an adenovirus encoding interferon-beta mediated strong antitumor responses in murine models with low but not high tumor burden. Our goals were to determine the mechanisms responsible for this loss of efficacy and to test the hypothesis that the combination of preoperative adenovirus encoding interferon-beta and surgical resection would be effective in treating bulky tumors. METHODS: Flank tumors of a mouse mesothelioma cell line were treated with adenovirus encoding interferon-beta or adenoviral vector encoding the bacterial protein beta-galactosidase. Cytotoxic T lymphocytes and tumor infiltration by T lymphocytes were measured. Tumors were surgically excised 72 hours later and tumor cells were injected in the contralateral flank to create a model of a metastatic focus. Tumor-free survival and distant metastatic disease were assessed. RESULTS: Immuno-gene therapy effectively treated small tumors (<200 mm(3)) but did not reduce the size of large (>800 mm(3)) flank tumors. Although treatment with adenovirus encoding interferon-beta resulted in the generation of tumor-neutralizing splenocytes in large tumors, the number of T cells visualized within the tumors was minimal. Tumors treated with adenovirus encoding interferon-beta (versus adenoviral vector encoding the bacterial protein beta-galactosidase or phosphate-buffered saline solution) prior to debulking increased long-term tumor-free survival and resulted in two- to sixfold smaller foci of implanted tumor cells at 2 weeks postoperatively. CONCLUSIONS: The use of adenovirus encoding interferon-beta or surgical debulking alone is ineffective in treating large tumors, but combining preoperative adenovirus encoding interferon-beta and surgical debulking significantly reduces tumor recurrence and improves long-term tumor-free survival. We postulate that adenovirus encoding interferon-beta amplifies the cytotoxic T-lymphocyte antitumor response, allowing elimination of residual tumor cells. PMID- 14752423 TI - Initial experience with the AbioCor implantable replacement heart system. AB - OBJECTIVE: We sought to evaluate the safety and efficacy of the first available totally implantable replacement heart (AbioCor Implantable Replacement Heart System) in the treatment of severe, irreversible biventricular heart failure in human patients. METHODS: Seven male adult patients with severe, irreversible biventricular failure (>70% thirty-day predicted mortality) who were not candidates for transplantation met all institutional review board study criteria and had placement of the AbioCor Implantable Replacement Heart. All were in cardiogenic shock despite maximal medical therapy, including inotropes and intra aortic balloon pumps. Mean age was 66.7 +/- 10.4 years (range, 51-79 years). Four of 7 patients had prior operations. Six had ischemic and one had idiopathic cardiomyopathy. All had 3-dimensional computer-simulated implantation of the thoracic unit that predicted adequate fit. At the time of the operation, the internal transcutaneous energy transfer coil, battery, and controller were placed. Biventriculectomy was then performed, and the thoracic unit was placed in an orthotopic position and attached to the atrial cuffs and outflow conduits with quick-connects. The flow was adjusted to 4 to 8 L/min. Central venous and left atrial pressures were maintained at 5 to 15 mm Hg. The device is powered through transcutaneous energy transfer. An atrial flow-balancing chamber is used to adjust left/right balance. The balance chamber and transcutaneous energy transfer eliminate the need for percutaneous lines. RESULTS: There was one intraoperative death caused by coagulopathic bleeding and one early death caused by an aprotinin reaction. There have been multiple morbidities primarily related to preexisting illness severity: 5 patients had prolonged intubation, 2 had hepatic failure (resolved in 1), 4 had renal failure (resolved in 3), and 1 each had recurrent gastrointestinal bleeding, acute cholecystitis requiring laparotomy, respiratory failure that resolved after 3 days of extracorporeal membrane oxygenation, and malignant hyperthermia (resolved). There were 3 late deaths: one caused by multiple systems organ failure (postoperative day 56), one caused by a cerebrovascular accident (postoperative day 142), and one caused by retroperitoneal bleeding and resultant multiple systems organ failure (postoperative day 151). This latter patient was not able to tolerate anticoagulation (no anticoagulation or antiplatelet therapy alone for 80% of the first 60 days) and had a transient ischemic attack on postoperative day 61 and a cerebrovascular accident on postoperative day 130. At autopsy, blood pumps were clean. The 2 patients who had large cerebrovascular accidents had thrombus on the atrial cage struts. These struts have been removed for future implants. There has been no significant hemolysis or device-related infections. The balance chamber has allowed for left/right balance in all patients (left atrial pressure within 5 mm Hg of right atrial pressure). Three patients have taken multiple (>50) trips out of the hospital, and 2 have been discharged from the hospital. Total days on support with the AbioCor are 759. CONCLUSION: The initial clinical experience suggests that the AbioCor might be effective therapy in patients with advanced biventricular failure. There have been no significant device malfunctions. Two of these patients have been discharged from the hospital. PMID- 14752424 TI - Equivalent midterm outcomes after off-pump and on-pump coronary surgery. AB - OBJECTIVE: Multiple reports demonstrate that off-pump surgery reduces the early morbidity associated with coronary artery bypass grafting. To determine if there are any differences in later outcomes, we compared midterm results of propensity matched patients who underwent off- and on-pump coronary artery bypass grafting. METHODS: From January 1997 to July 2000, 481 patients underwent off-pump coronary artery bypass grafting and 3231 underwent on-pump coronary artery bypass grafting. Propensity matching was used to match 406 patients from each group. Previously, the propensity-matched off-pump patients were found to have had significantly fewer bypass grafts. These 812 patients were followed for time related events, including death, myocardial infarction, percutaneous coronary intervention, coronary reoperation, and the combined end point of all-cause mortality, myocardial infarction, and all coronary reintervention. Follow-up was 95% complete. RESULTS: At 4 years, survival was 87.5% after off-pump and 91.2% after on-pump coronary artery bypass grafting (P =.2); freedom from myocardial infarction was 92.6% and 95.7% (P =.7), respectively; freedom from percutaneous coronary intervention was 94.3% and 95.5% (P =.9), respectively; freedom from coronary reoperation was 98.1% and 99.0% (P =.4), respectively; and freedom from the combined end point of all-cause mortality, myocardial infarction, and coronary reintervention was 75.2% and 82.9% (P =.14), respectively. CONCLUSIONS: Off-pump and on-pump coronary artery bypass grafting results in equivalent midterm outcomes. Fewer bypass grafts in the off-pump patients did not decrease survival or increase ischemic events at 4 years. PMID- 14752426 TI - Composite arterial grafts versus conventional grafting for coronary artery bypass grafting. AB - OBJECTIVES: Composite arterial grafts for coronary artery bypass grafting surgery allow complete arterial revascularization but are limited by the inflow of a single internal thoracic artery supplying all the grafted vessels. We reviewed the safety of composite arterial grafts using either bilateral internal thoracic arteries or a single internal thoracic artery and radial artery. METHODS: From January 1999 to July 2002, 402 consecutive patients receiving composite grafts only were compared to a control group of patients (n = 542) undergoing coronary artery bypass grafting with internal thoracic artery and saphenous veins operated upon by the same surgeons. Two different statistical approaches were used to compare groups in this retrospective analysis. First, propensity score analysis with greedy matching technique was used to match patients from each group. Second, a multivariate analysis was performed looking at a combined patient outcome of death, intra-aortic balloon counterpulsation utilization, myocardial infarction, stroke, and prolonged ventilation on all patients in both groups. RESULTS: After matching by propensity score, the major clinical outcomes in composite arterial (n = 249) and control (n = 249) groups were found to be similar. The in-hospital mortality in the composite group was 1.2% as compared with 0.4% in matched patients (P =.62). However, patients in the composite group were found to have a significantly longer pump time (P <.0001), longer clamp time (P <.0001), increased incidence of prolonged mechanical ventilation (12.8% vs 4.8%; P =.002), and higher incidence of combined morbidity outcome (13.6% vs 6.4%; P =.007) as compared with matched patients. Multivariable analysis showed that composite arterial grafting was an independent predictor of the combined morbidity outcome with an odds ratio of 2.1 (1.2-3.7). CONCLUSIONS: These findings suggest that composite arterial grafting may be associated with an increase in risk-adjusted patient morbidity when compared with a conventional coronary artery bypass grafting group, although a mortality difference was not demonstrable. PMID- 14752425 TI - Late incidence and predictors of persistent or recurrent heart failure in patients with aortic prosthetic valves. AB - BACKGROUND: We examined factors associated with persistent or recurrent congestive heart failure after aortic valve replacement. METHODS: Patients who underwent aortic valve replacement with contemporary prostheses (n = 1563) were followed up with annual clinical assessment and echocardiography. The effect of demographic, comorbid, and valve-related variables on the composite outcome of New York Heart Association class III or IV symptoms or congestive heart failure death after surgery was evaluated with stratified log-rank tests, Cox proportional hazard models, and logistic regression. Factors associated with all cause death were also examined. Prediction models were bootstrapped 1000 times. RESULTS: Total follow-up was 6768 patient-years (mean, 4.3 +/- 3.3 years; range, 60 days to 17.1 years). Freedom from congestive heart failure or congestive heart failure death was 98.6% +/- 0.3%, 88.6% +/- 1.0%, 73.9% +/- 2.3%, and 45.2% +/- 8.5% at 1, 5, 10, and 15 years, respectively. Age, preoperative New York Heart Association class, left ventricular grade, atrial fibrillation, coronary artery disease, smoking, and redo status predicted congestive heart failure after surgery (all P <.05). Larger prosthesis size and effective orifice area, both absolute and indexed for body surface area, were independently associated with freedom from congestive heart failure. Increased transprosthesis gradients were predicted by prosthesis-patient mismatch and were associated with congestive heart failure after surgery. Mismatch defined as an effective orifice area/body surface area of 0.80 cm(2)/m(2) or less was a significant predictor of congestive heart failure events after surgery, but mismatch defined as an effective orifice area/body surface area of 0.85 cm(2)/m(2) or less was not. Small prosthesis size and mismatch were not significantly associated with all-cause mortality. CONCLUSIONS: These analyses identify independent predictors of congestive heart failure symptoms and congestive heart failure death late after aortic valve replacement and indicate that prosthesis size has a significant effect on this cardiac end point, but not on overall survival after aortic valve replacement. PMID- 14752427 TI - Comparison of coronary bypass surgery with and without cardiopulmonary bypass in patients with multivessel disease. AB - BACKGROUND: Coronary artery bypass grafting can now be performed with or without cardiopulmonary bypass. Our objective was to determine whether off-pump coronary artery bypass grafting is associated with better early outcomes compared with conventional coronary artery bypass grafting. METHODS: In 4 centers with off-pump coronary surgery experience, a retrospective analysis of all coronary artery bypass grafting in a 3-year period was performed. Groups were compared to determine selection criteria, mortality, and morbidity, then computer-matched by propensity score to control for selection bias. Multivariate logistic regression identified risk factors predictive of mortality. Specific subgroups most likely to benefit were identified. RESULTS: In all, 17,401 isolated coronary artery bypass grafts were performed, 7283 (41.9%) off-pump coronary artery bypass grafts and 10,118 (58.1%) conventional coronary artery bypass with cardiopulmonary bypass. Factors determining selection of patients for off-pump coronary artery bypass grafting included female gender (55.5% vs 44.5%), preexisting renal failure (57.0% vs 43.0%), and reoperations (52.6% vs 47.4%). Operative mortality was 2.8%; off-pump coronary artery bypass grafting versus conventional coronary artery bypass with cardiopulmonary bypass (1.9% vs 3.5%, P <.001) had the same predicted risk. Of the patients with multivessel disease, 11,548 were matched by propensity scoring. Mortality was significantly less in the off-pump coronary artery bypass grafting group (2.8% vs 3.7%, P <.001). By multivariate logistic regression analysis of the matched sample, predictors for mortality were female gender (odds ratio 1.83, confidence interval 1.37-2.44), preexisting renal failure (odds ratio 2.85, confidence interval 2.64-4.95), history of stroke (odds ratio 1.74, confidence interval 1.08-2.80), previous coronary artery bypass grafting surgery (odds ratio 4.22, confidence interval 2.92-6.09), use of cardiopulmonary bypass (odds ratio 2.08, confidence interval 1.52-2.83), and recent myocardial infarction (odds ratio 2.31, confidence interval 1.68-3.22). Cardiopulmonary bypass was predictive of mortality in reoperations, female patients, and patients aged >or= 75 years. Off-pump coronary artery bypass grafting was associated with less morbidity, including reductions in blood transfusion (32.6% vs 40.6%, P <.001), stroke (1.4% vs 2.1%, P =.002), renal failure (2.6% vs 5.2%, P <.001), pulmonary complications (4.1% vs 9.5%, P <.001), reoperation (1.7% vs 3.2%, P <.001), atrial fibrillation (21.1% vs 24.99%, P <.001), and gastrointestinal complications (3.6% vs 4.8%, P =.02). CONCLUSION: In 4 centers with beating-heart operation experience, there is an overall early benefit in off-pump surgery, especially in patients traditionally considered at high risk for coronary artery bypass grafting. PMID- 14752428 TI - Coronary artery bypass grafting: are risk models developed from on-pump surgery valid for off-pump surgery? AB - OBJECTIVE: This study was undertaken to test whether risk models developed from on-pump coronary artery bypass grafting are valid for assessing the risk for off pump coronary artery bypass grafting. METHODS: From January 1997 through June 2002, a total of 12,845 patients underwent isolated coronary artery bypass grafting procedures in Providence Health System hospitals. Of these, 1782 operations (14%) were performed without cardiopulmonary bypass. An operative mortality risk model was derived from on-pump data with logistic regression. This model and two other external risk models developed from on-pump data were then applied to patients undergoing off-pump coronary artery bypass grafting to test the model adequacy. RESULTS: Good model discrimination and calibration were obtained from all three models. CONCLUSION: Operative mortality risk models developed from on-pump coronary artery bypass grafting can be used to assess the risk for off-pump coronary artery bypass grafting. PMID- 14752429 TI - Left internal thoracic artery-radial artery composite grafts as the technique of choice for myocardial revascularization in elderly patients: a prospective randomized evaluation. AB - OBJECTIVES: The technique of choice for myocardial revascularization in elderly patients remains a debated issue. We evaluated the potential advantages of the use of left internal thoracic artery-radial artery composite grafts compared with conventional coronary artery bypass grafts in elderly patients. METHODS: We prospectively enrolled 160 patients aged more than 70 years scheduled to undergo isolated myocardial revascularization. Patients were assigned at random to group 1, 80 patients undergoing total arterial revascularization (left internal thoracic artery on left anterior descending coronary artery plus radial artery), or group 2, 80 patients undergoing standard coronary artery bypass graft surgery (left internal thoracic artery on left anterior descending coronary artery plus saphenous veins). The radial artery was used in all cases as a composite Y-graft. RESULTS: Preoperative characteristics and risk factors (EuroSCORE: group 1 = 7.9 vs group 2 = 8.1), number of grafted coronary vessels (group 1 = 2.4 vs group 2 = 2.5), aortic crossclamping time (group 1 = 37 +/- 7 minutes vs group 2 = 38 +/- 7 minutes), ventilation time (group 1 = 22 +/- 12 hours vs group 2 = 23 +/- 11 hours), intensive care unit stay (group 1 = 39 +/- 10 hours vs group 2 = 40 +/- 9 hours), and hospital mortality (group 1 = 3.8% vs group 2 = 5%) were comparable between the groups. Comparison between the 2 groups in terms of early postoperative complications showed a higher incidence of cerebrovascular accidents in group 2 (group 1 = 0 patients vs group 2 = 4 patients, 5%). At a mean follow-up of 16 +/- 3 months, patients in group 1 showed superior clinical results with a lower incidence of graft occlusion (group 1 = 2 vs group 2 = 11; P =.06) and angina recurrence (group 1 = 2 patients vs group 2 = 12 patients; P =.03). Multivariate analysis identified saphenous vein grafts as independent predictors for graft occlusion and angina recurrence. CONCLUSIONS: Left internal thoracic artery-radial artery composite grafts proved to be a safe procedure in elderly patients. It improved the clinical outcome, providing a significantly higher graft patency rate and a lower incidence of late cardiac events. PMID- 14752430 TI - Automated distal coronary bypass with a novel magnetic coupler (MVP system). AB - OBJECTIVE: We sought to assess the feasibility of performing sutureless distal coronary artery bypass anastomoses with a novel magnetic coupling device. METHODS: From May 2000 to April 2001, single-vessel side-to-side coronary artery bypass grafting on a beating heart was performed in 39 domestic white pigs (35-60 kg) without the use of mechanical stabilization, shunts, or perfusion bridges. Animals were divided into 2 groups. Seventeen pigs underwent right internal thoracic artery to right coronary artery bypass grafting through a median sternotomy (group 1) with a novel magnetic vascular positioning system (MVP system; Ventrica, Inc, Fremont, Calif). Twenty-two pigs underwent left internal thoracic artery to left anterior descending artery grafting with the MVP anastomotic device through a left anterior minithoracotomy (group 2). This system consists of 2 pairs of elliptical magnetic implants and a deployment device. One pair of magnets forms the anastomotic docking port within the graft; the other pair forms an identical anastomotic docking port within the target vessel. The anastomosis is created when the 2 docking ports magnetically couple. Anastomotic patency was evaluated by means of angiography during the first postoperative week and at 1 month. Histologic studies were performed at different time points as late as 6 months. RESULTS: Right internal thoracic artery to right coronary artery anastomoses and left internal thoracic artery to left anterior descending artery anastomoses were successfully performed with the system in all animals. The self-adherent and self-aligning properties of the implants allowed for immediate and secure approximation of the arteries (total anastomotic time between 2-3 minutes). Anastomoses were constructed without a stabilization platform. Five nondevice-related deaths occurred postoperatively. One-week angiography, performed in 35 surviving animals, showed a patent graft and anastomosis in all cases. The patency rate at 1 month was 97% (33/34). Histologic studies as late as 6 months demonstrated neointimal coverage of the magnets without any significant luminal obstruction. Histology also confirmed the presence of viable tissue between magnets. CONCLUSION: The MVP anastomotic system uses magnetic force to create rapid and secure distal coronary artery anastomoses, which might facilitate minimally invasive and totally endoscopic coronary artery bypass surgery. PMID- 14752431 TI - Primary repair of pulmonary atresia with ventricular septal defect and major aortopulmonary collaterals: a useful approach. AB - BACKGROUND: The ultimate goal of surgical therapy for pulmonary atresia with ventricular septal defect and major aortopulmonary collateral arteries is to create unobstructed and separate in series pulmonary and systemic circuits. Our preference has been a 1-stage complete unifocalization technique, avoiding collateral anastomosis with either the native pulmonary arteries or other aortopulmonary collateral vessels. METHODS AND RESULTS: Since 1998, 5 patients (median age 29.6 months) with pulmonary atresia, ventricular septal defect, and major aortopulmonary collateral arteries have undergone surgical correction, consisting of (1) exclusion of a descending thoracic aortic segment from which all major aortopulmonary collateral arteries originate, and (2) connection of this aortic segment to the native pulmonary artery using an interposition polytetrafluoroethylene conduit. The ventricular septal defect was closed in all patients, and the right ventricle was connected to the unifocalized pulmonary artery with a valved conduit. All patients survived the operation. Two patients required reexploration for postoperative bleeding. One patient remained on mechanical ventilation for 17 days due to a pulmonary infection. During follow-up (12-21 months), no patient required additional interventions. The postoperative right ventricular/left ventricular pressure ratio was 0.55 median. No significant stenosis within the reconstructed pulmonary circuit was identified. All patients remain free of symptoms, requiring no medications. CONCLUSION: Intracardiac repair of pulmonary atresia with ventricular septal defect and major aortopulmonary collateral arteries can be accomplished by a midline 1-stage repair including complete unifocalization of all pulmonary blood supply without individual collateral anastomosis in selected patients. This approach offers a convenient and satisfactory surgical option. PMID- 14752432 TI - Is surgical intervention still indicated in recurrent aortic arch obstruction? AB - BACKGROUND: Introduction of balloon dilatation has become the standard treatment for recurrent aortic arch obstruction and has changed the therapeutic approach to patients with this disorder. OBJECTIVES: Whether all patients with recurrent aortic arch obstruction are candidates for balloon dilatation remains unanswered. In addition, only few reports have tried to compare the results between patients undergoing balloon dilatation or redo operations. METHODS: Since 1983, 97 patients underwent reintervention for recurrent aortic arch obstruction (42 dilations and 55 reoperations). Eight had immediate unsuccessful dilatation and were shifted to the surgical group (n = 63). The median age at reintervention was 21.7 months (10 days-45 years), and the median delay was 13.6 months (7 days-17 years). Anatomy of the aortic arch oriented the surgical approach to treat arch hypoplasia. It could be performed through a left thoracotomy in 52 patients, with extended end-to-end anastomosis in 34 patients, subclavian flap repair in 9 patients, conduit insertion in 6 patients, and patch enlargement in 3 patients. More recently, an anterior approach with cardiopulmonary bypass without circulatory arrest was applied to enlarge the patch in all the aortic arches. RESULTS: There was one early death in the surgical intervention group and 2 late deaths in the dilation group. Major complications and recurrence were higher in the dilated group (4 vs 0, P <.01, and 14 vs 5, P <.0004, respectively). At a mean follow-up of 11.8 +/- 4.1 years in the surgical intervention group and 7.5 +/- 2.5 years in the dilated group, systemic hypertension was normalized in all but 5 patients in the surgical intervention group and 6 patients in the dilated group. CONCLUSION: Reoperation for recurrent aortic arch obstruction can be performed safely, with low rates of mortality and morbidity. This approach should be considered versus balloon angioplasty, especially in patients older than 4 years and in the presence of aortic arch hypoplasia. PMID- 14752433 TI - Arterial switch with full-flow cardiopulmonary bypass and limited circulatory arrest: neurodevelopmental outcome. AB - OBJECTIVES: Neonatal cardiac surgery has been associated with unfavorable neurodevelopmental events. We investigated a patient cohort operated on predominantly with full-flow cardiopulmonary bypass (150 mL x kg(-1) x min(-1), alpha-stat, alpha-blockade, median arrest = 6 minutes, temperature of 22 degrees C) as the major support strategy for neonatal arterial switch operations (transposition of the great arteries and intact ventricular septum). METHODS: Seventy-four patients and "best-friend" control subjects were assessed 109 months (range, 48-166 months) postoperatively with general medical and neurologic evaluation, IQ testing, formal movement scores, and detailed parent-teacher behavioral-social reports. Fetal, neonatal, and perioperative data were collated. RESULTS: The prevalence of perioperative seizures was 6.8% (4/5 cases occurring preoperatively). The incidence of all perioperative neurologic abnormalities was 20%. Patients who had a neurologic event were (as a group) older at the time of operation and had a lower arterial blood pH before the operation. Selected perioperative factors (not related directly to cardiopulmonary bypass variables) predicted early (before discharge) neurologic outcome in a multivariate model. At late assessment, patients were more likely than control subjects to have a mild neurologic abnormality (P = 0.002). Full-scale IQ scores (Wechsler Preschool and Primary Scale of Intelligence and Wechsler Intelligence Scale for Children-Third Edition) were higher in control subjects (101.9 [SD = 13] vs 108.6 [SD = 12], P =.0007), with both groups having scores greater than the population-based test means. Full-scale IQ scores related most significantly to years of paternal education (beta = 1.51, P =.0078) but were also influenced by perioperative neurologic abnormalities, birth weight, and circulatory arrest time. Patients had higher motor impairment scores (Movement Assessment Battery) than control subjects (P =.0004). Parents (Achenbach Child Development Checklist) assigned higher total social-behavioral competence scores to control subjects (P =.05). Teachers (Achenbach Teacher Report Form) suggested that patients were more likely to be perceived as having various speech and expressive language problems, as well as minor behavioral problems. CONCLUSION: With the perioperative strategies used, not all survivors can be considered (neurodevelopmentally) normal at late follow-up, although the risk of important impairment is low. Perioperative events might have long-term prognostic value. On the basis of this study and published data regarding other strategies, continued application of full-flow cardiopulmonary bypass is justified, with the proviso that further investigation is required. PMID- 14752434 TI - Changes in cerebral and somatic oxygenation during stage 1 palliation of hypoplastic left heart syndrome using continuous regional cerebral perfusion. AB - OBJECTIVES: Stage 1 palliation of hypoplastic left heart syndrome requires the interruption of whole-body perfusion. Delayed reflow in the cerebral circulation secondary to prolonged elevation in vascular resistance occurs in neonates after deep hypothermic circulatory arrest. We examined relative changes in cerebral and somatic oxygenation with near-infrared spectroscopy while using a modified perfusion strategy that allowed continuous cerebral perfusion. METHODS: Nine neonates undergoing stage 1 palliation for hypoplastic left heart syndrome had regional tissue oxygenation continuously measured by frontal cerebral and thoraco lumbar (T10-L2) somatic (renal) reflectance oximetry probes (rSO(2), INVOS; Somanetics, Troy, Mich). Surgery was accomplished using cardiopulmonary bypass with whole-body cooling (18 degrees C-20 degrees C) and regional cerebral perfusion through the innominate artery at flow rates guided by estimated minimum flow requirements and measured rSO(2) during reconstruction of the aortic arch. Data were logged at 1-minute intervals and analyzed using repeated measures analysis of variance. RESULTS: A total of 3176 minutes of data were analyzed. Prebypass cerebral rSO(2) was 65.4 +/- 8.9, and somatic rSO(2) was 58.9 +/- 12.4 (P <.001, cerebral vs somatic). During regional cerebral perfusion, cerebral rSO(2) was 80.7 +/- 8.6, and somatic rSO(2) was 41.4 +/- 7.1 (P <.001). Postbypass cerebral rSO(2) was 53.2 +/- 14.9, and somatic rSO(2) was 76.4 +/- 7.7 (P <.001). The risk of cerebral desaturation was significantly increased after cardiopulmonary bypass. CONCLUSIONS: Cerebral oxygenation was maintained during regional cerebral perfusion at prebypass levels with deep hypothermia. However, after rewarming and separation from cardiopulmonary bypass, cerebral oxygenation was lower compared with prebypass or somatic values. These results indicate that cerebrovascular resistance is increased after deep hypothermic cardiopulmonary bypass, even with continuous perfusion techniques, placing the cerebral circulation at risk postoperatively. PMID- 14752435 TI - Intraoperative device closure of perimembranous ventricular septal defects without cardiopulmonary bypass: preliminary results with the perventricular technique. AB - OBJECTIVE: In infants undergoing closure of perimembranous ventricular septal defects, cardiopulmonary bypass remains one of the factors that prolongs hospital stay and morbidity. A new technique was used to close the defects under echocardiographic guidance without cardiopulmonary bypass to prevent the deleterious effects of bypass. METHODS: Recently, the Amplatzer membranous ventricular septal defect device (AGA Medical Corp, Golden Valley, Minn) was introduced. The device has a double-disc design with a short connecting waist. The left ventricular disc has an eccentric design to prevent encroachment on the aortic valve leaflets. Eight Yucatan miniature pigs with naturally occurring perimembranous ventricular septal defects underwent closure of the defect in the operating room by using the perventricular technique. After median sternotomy, a purse-string suture was placed on the free wall of the right ventricle. An angiocatheter was advanced in the right ventricle, and through the catheter, a wire was advanced from the right ventricle through the ventricular septal defect into the left ventricle. A delivery sheath and the dilator were advanced over the wire. The wire and catheter were removed, and an appropriately sized Amplatzer membranous device was advanced through the sheath. The device was deployed under echocardiographic guidance with the heart beating. RESULTS: The procedure was successful in all animals. There was no incidence of device embolization, heart block, or aortic insufficiency. Angiograms at 3 and 6 months revealed no residual defects and no aortic insufficiency. Pathologically, the devices were completely endothelialized when examined grossly. CONCLUSIONS: The perventricular technique appears to be excellent for closure of perimembranous ventricular septal defects in the operating room. The technique might be feasible in smaller babies, who are high-risk candidates for closure in the catheterization laboratory. Cardiopulmonary bypass and prolonged hospital stay are avoided. PMID- 14752436 TI - Blood group incompatibility and accelerated homograft fibrocalcifications. AB - OBJECTIVE: Cryopreserved valved homograft has become the conduit of choice for right ventricular outflow tract reconstruction in pediatric cardiac surgery. Aortic homografts have been frequently used in pulmonary position, but accelerated aortic homograft fibrocalcification may occur. Blood group incompatibility between receiver and homograft donor may play a central role in this context. METHODS: Between 1993 and 2000, 59 children (mean age 6.4 +/- 4.4 years) received cryopreserved valved homografts for right ventricular outflow tract reconstruction and were followed from 2 to 10 years clinically, with echocardiography and chest radiography for detection of development of homograft calcifications. Seventeen patients were 3 years or younger. Fifty aortic (85%) and 9 pulmonary homografts were all used in pulmonary position. Thirty-three patients (56%) had the same blood group (ABO) as the homograft donor (iso group), and 26 were blood group-incompatible (non-iso group). RESULTS: No deaths occurred during follow-up. Six patients (10.2%) required homograft replacement because of severe fibrocalcifications, and another 3 showed moderate homograft calcifications (5.1%) at last examination. Freedom from moderate to severe homograft calcification at 8 years (Kaplan-Meier) was 95.2% for the iso group and 72.9% for the non-iso group (P <.0001). Homograft calcifications occurred within 2 years of implantation in 6/9 patients (67%) in the non-iso group. CONCLUSIONS: Blood group incompatibility between receiver and homograft donor seems to play an important role in the development of accelerated fibrocalcifications in cryopreserved homografts, particularly in the very young (3 years old or younger). Blood group compatibility should therefore be respected to avoid accelerated homograft fibrocalcifications. PMID- 14752437 TI - The influence of pulmonary artery morphology on the results of operations for major aortopulmonary collateral arteries and complex congenital heart defects. AB - OBJECTIVE: Congenital heart defects with major aortopulmonary collateral arteries show marked variability in the size and distribution of native pulmonary arteries. We sought to classify the size and distribution of native pulmonary arteries and to determine their influence on surgical outcome. METHODS: Between 1989 and 2002, 164 patients underwent surgical intervention for congenital heart defects with major aortopulmonary collateral arteries (median age, 10 months). Three patterns of native pulmonary arteries were identified: intrapericardial native pulmonary arteries present (group I); confluent intrapulmonary native pulmonary arteries without intrapericardial native pulmonary arteries (group II); and nonconfluent intrapulmonary native pulmonary arteries (group III). Thirty seven (23%) patients had single-stage and 76 (47%) patients had multistage complete repair. Thirty (18%) patients await septation, and 8 (5.0%) patients are not septatable. Follow-up is 98% complete (median follow-up, 5.8 years). RESULTS: In the 164 patients there were 15 (9.1%) early and 12 (7.3%) late deaths. Early mortality after complete repair was 4.4% (n = 5). Actuarial survival was 90% +/- 3% and 85% +/- 4% at 1 and 10 years, respectively. Actuarial freedom from surgical or catheter reintervention in septated patients was 77% +/- 4% and 45% +/- 8% at 1 and 10 years, respectively. On multivariate analysis, the morphology of the native pulmonary arteries was the only factor that influenced actuarial survival after complete repair (P =.04). Group III had the highest risk of death after septation (P =.008). Group II fared better than group III after the initial operation (P <.05). CONCLUSIONS: Current classifications of congenital heart defects with major aortopulmonary collateral arteries are based on the presence or absence of intrapericardial pulmonary arteries. We have identified a subgroup without intrapericardial native pulmonary arteries but with confluent intrapulmonary native pulmonary arteries. This group has a better outcome than those with nonconfluent intrapulmonary native pulmonary arteries. PMID- 14752438 TI - Prosthetic graft remnant-related pseudoaneurysm after left ventricular assist device explantation: a case report. PMID- 14752439 TI - Off-pump technique for insertion of a HeartMate Vented Electric left ventricular assist device. PMID- 14752440 TI - Management of aortic insufficiency in patients with left ventricular assist devices: a simple coaptation stitch method (Park's stitch). PMID- 14752442 TI - Use of a proximal anastomotic device in coronary artery bypass surgery: a word of caution. PMID- 14752441 TI - Failure of four bovine pericardial mitral prostheses. PMID- 14752443 TI - Apicoaortic conduit in a patient with severe hemolysis after three aortic valve replacements. PMID- 14752444 TI - Selective perfusion of preoperatively identified artery of Adamkiewicz during repair of thoracoabdominal aortic aneurysm. PMID- 14752445 TI - Ventricular septal defect caused by impact from a horseshoe. PMID- 14752446 TI - Selective carotid cannulation at the neck: a satisfactory option for reoperation for aneurysms of the ascending aorta and arch. PMID- 14752447 TI - Aortic arch aneurysm with dissection in Cushing syndrome. PMID- 14752448 TI - Maintenance of hemodynamic stability during pericardiectomy with the Starfish 2 Heart Positioner. PMID- 14752449 TI - Pulmonary venous pathway obstruction from recurrent restriction at atrial septum late after Fontan procedure. PMID- 14752450 TI - Complex double-outlet right ventricle repair in a neonate with complete tracheal agenesis. PMID- 14752451 TI - Formation of a stenotic fibrotic membrane at the distal anastomosis of bovine jugular vein grafts (Contegra) after right ventricular outflow tract reconstruction. PMID- 14752452 TI - Cardiac resynchronization therapy in a patient with single ventricle and intracardiac conduction delay. PMID- 14752454 TI - Mediastinal growing teratoma syndrome after cisplatin-based chemotherapy and radiotherapy for intracranial germinoma. PMID- 14752453 TI - Intrasternal respiratory epithelium-lined foregut cyst. PMID- 14752455 TI - A rare case of hemangioma arising from the azygos vein: Informative procedure with endobronchial ultrasonography. PMID- 14752456 TI - Gastrobronchial fistula and anastomotic esophagogastric stenosis after esophagectomy for esophageal carcinoma. PMID- 14752457 TI - Bipulmonary transplants with lungs obtained from two non-heart-beating donors who died out of hospital. PMID- 14752458 TI - Off-pump versus conventional coronary artery bypass grafting: randomized studies. PMID- 14752459 TI - Abdominal tumors with cavoatrial extension. PMID- 14752460 TI - Partial left ventriculectomy in patients with neoplasms and severe heart failure who are not candidates for cardiac transplantation. PMID- 14752461 TI - Aortic insufficiency in patients with Marfan syndrome: a surgical dilemma. PMID- 14752462 TI - Amiodarone for postoperative atrial fibrillation. PMID- 14752463 TI - Endotoxemia and cardiac function. PMID- 14752464 TI - Ascending aorta cannulation in type A dissection. PMID- 14752466 TI - Assessment of coronary flow reserve and microcirculation: a clinical perspective. PMID- 14752467 TI - Sensitivity of acute rest myocardial perfusion imaging for identifying patients with myocardial infarction based on a troponin definition. AB - BACKGROUND: Myocardial perfusion imaging (MPI) is often used to identify low-risk chest pain patients who have myocardial infarction (MI). A recent recommendation is that patients with increased troponin levels be diagnosed as having MI. The sensitivity and characteristics of patients who have elevated troponin levels who also underwent early MPI are unknown. METHODS AND RESULTS: Patients considered at low risk for MI underwent rest gated tomographic MPI and serial marker assessment as part of a standard chest pain evaluation protocol. Patients with cardiac troponin I (cTnI) elevations were analyzed further for this study. MPI results were considered positive if there was a perfusion defect in association with abnormal wall motion or thickening. Short-axis images were divided into 17 segments and graded on a 4-point scale (0, normal; 3, high-grade or absent perfusion), and a summed rest score was derived. Of the 319 patients who had MPI and cTnI elevations, 78 had negative MPI results (sensitivity, 75%). Patients with negative MPI results had lower peak creatine kinase (CK)-MB values (15 +/- 25 ng/mL vs 45 +/- 78 ng/mL, P <.0001) and higher ejection fractions (56% +/- 15% vs 47% +/- 13%, P <.0001) and were less likely to have significant disease (55% vs 72%, P =.04) than those with positive MPI results. Increasing summed rest score was associated with larger MIs as estimated by peak CK and CK-MB values. CONCLUSIONS: Patients with negative MPI results have smaller MIs and less extensive coronary disease. MPI and cTnI offer complementary data for assessing patients with possible MI. PMID- 14752468 TI - Adenosine stress myocardial perfusion tomographic imaging in patients with significant aortic stenosis. AB - BACKGROUND: Myocardial perfusion scintigraphy has been used by some investigators for the diagnosis of coronary artery disease (CAD) in patients with mild, moderate, and moderate to severe aortic stenosis, with various results. The aim of this study was to assess the safety and diagnostic accuracy of adenosine stress myocardial perfusion scintigraphy (adenosine single photon emission computed tomography [Ad-SPECT]) for the detection of CAD in patients with significant aortic stenosis. METHODS AND RESULT: The study included 75 patients with significant aortic stenosis (maximal instantaneous aortic valve gradient >80 mm Hg [range, 81-149 mm Hg] and aortic valve area <0.75 cm2). All patients underwent Ad-SPECT after a 6-minute infusion of adenosine (140 microg/kg body weight per minute). At the third minute of adenosine infusion, a bolus of 3 mCi thallium 201 was injected, and SPECT acquisition was obtained immediately after completion of adenosine infusion. Coronary angiography was performed in all patients. No major complications during adenosine infusion were observed. All unpleasant symptoms lasted for only a few seconds and did not necessitate cessation of the test. Concerning the angiographically diagnosed CAD, we found that Ad-SPECT showed a sensitivity of 88.6%, a specificity of 72.5%, a positive predictive value of 73.8%, a negative predictive value of 87.8%, and a diagnostic accuracy of 80%. CONCLUSIONS: Ad-SPECT is a moderately accurate method for detecting the presence or absence of CAD in patients with severe aortic stenosis. However, further modification of this method is required before it can supplant cardiac catheterization in the preoperative evaluation of patients with severe aortic stenosis. PMID- 14752469 TI - Clinical significance of apical thinning after attenuation correction. AB - BACKGROUND: Apical thinning and other image changes at the apex have been described after attenuation correction of myocardial perfusion single photon emission computed tomography (SPECT) studies, but their clinical significance is unknown. METHODS AND RESULTS: We studied 102 subjects from a multicenter trial of attenuation correction, 46 with angiographic coronary artery disease and 56 normal subjects. We graded the presence and magnitude (on a 4-point scale) of apical thinning (decrease in wall thickness, best assessed in the vertical long axis view) in both noncorrected and attenuation-corrected images. In attenuation corrected images, apical thinning of any degree was present in 78% of the abnormal patients and 63% of the normal subjects (P = not significant [NS]). However, moderate or severe apical thinning was present in 30% of the abnormal patients compared with 5% of the normal subjects (relative risk = 2.2, P <.001). In noncorrected images, apical thinning of any degree was present in 87% of the abnormal patients and 71% of the normal subjects (P = NS). However, moderate or severe apical thinning was present in 28% of the abnormal patients compared with 4% of the normal subjects (relative risk = 2.3, P <.001). CONCLUSION: The presence of mild apical thinning is common in both noncorrected and attenuation corrected SPECT images and does not imply coronary artery disease. Moderate or severe apical thinning is 7 times more common in patients than in normal subjects, but it is relatively uncommon and thus is not a generally useful clinical tool. PMID- 14752470 TI - Decreased septal wall thickening in patients with left bundle branch block. AB - OBJECTIVES: Septal wall motion abnormalities are frequently observed in patients with left bundle branch block (LBBB). However, septal wall thickening in LBBB patients has not been thoroughly evaluated. METHODS AND RESULTS: To investigate the relationship between septal wall motion and wall thickening, we studied 31 normal control subjects, 24 LBBB patients with normal wall motion (LBBB-NWM), and 24 LBBB patients with septal dyssynchrony (LBBB-SDS), all with a low likelihood (<15%) of coronary artery disease. The septal and lateral quadrants of the left ventricle were analyzed in stress 8-frame gated technetium 99m sestamibi tomograms. The percent wall thickening was calculated by use of a 25-segment polar map with the p-FAST software program by two independent methods: the regional count density increase from end diastole to end systole (CD method) and the geometric increase in the distance between the 50% thresholded endocardial and epicardial borders from end diastole to end systole (GD method). In addition, the ratio of septal/lateral percent wall thickening was calculated. The relative septal wall thickening in the entire LBBB population was decreased as compared with the normal control subjects (0.35 +/- 0.37 vs 0.81 +/- 0.17, P <.001). Decreased wall thickening was observed in not only LBBB-SDS patients but also to a lesser degree in LBBB-NWM patients (0.12 +/- 0.35, P <.001; 0.57 +/- 0.24, P =.005, respectively). This abnormality was most apparent when the CD method was used. CONCLUSIONS: Septal wall thickening is decreased in patients with LBBB even with normal wall motion. LBBB per se may compromise septal wall thickening, and dyssynchronous wall motion results in further deterioration of wall thickening. PMID- 14752472 TI - Day-to-day variability of global left ventricular functional and perfusional measurements by quantitative gated SPECT using Tc-99m tetrofosmin in patients with heart failure due to coronary artery disease. AB - BACKGROUND: Although myocardial gated single photon emission computed tomography (SPECT) is routinely used for functional measurements in patients with coronary artery disease (CAD) and heart failure, day-to-day variability of left ventricular ejection fraction (LVEF), left ventricular (LV) volumes, and global perfusion scoring has not yet been investigated. METHODS AND RESULTS: In 20 consecutive patients with CAD and an LVEF lower than 40% who routinely underwent a resting tetrofosmin gated SPECT study, we performed an additional gated SPECT study at rest 1 to 5 days later under the same circumstances. LV volumes and LVEF were calculated from the gated SPECT data by commercially available software (QGS). Myocardial perfusion was scored visually by use of a 20-segment, 5-point scoring method. For global LV function and perfusion, agreement between data was investigated by use of Bland-Altman plotting. The 95% limits of agreement found by Bland-Altman analysis were -0.9% +/- 6.0% for LVEF, 3 +/- 20 mL for LV end diastolic volume, and 4 +/- 20 mL for LV end-systolic volume. CONCLUSION: In CAD patients with an LVEF lower than 40%, day-to-day variability of measurements of global myocardial function and perfusion is quite similar to interobserver and intraobserver variability. Day-to-day variability of global LV functional parameters obtained by gated cardiac SPECT is fairly small, which indicates that myocardial gated SPECT can be used in daily clinical practice to determine changes in global LV function and perfusion over time in patients with diminished LV function. PMID- 14752471 TI - Absolute quantitation of left ventricular wall and cavity parameters using ECG gated PET. AB - BACKGROUND: Electrocardiography (ECG)-gated scintigraphy demonstrates promising results for the simultaneous assessment of myocardial glucose metabolism and contractile function. In this study a method was evaluated for absolute quantitation of left ventricular wall and cavity parameters with the use of fluorine 18 fluorodeoxyglucose (FDG) ECG-gated positron emission tomography (PET). METHODS AND RESULTS: A previously developed 2-dimensional mathematical model was implemented for computer-automated identification of endocardial and pericardial borders. The accuracy and precision were tested in a heart phantom and in healthy subjects. Twelve healthy men aged 64 +/- 8 years were studied by use of cine magnetic resonance imaging (MRI) and ECG-gated FDG-PET during euglycemic glucose-insulin clamp. At increasing image noise levels, the estimated cavity volume of the heart phantom was within 2 mL of the actual volume, and no significant difference was found between actual and estimated wall thicknesses. Endocardial wall motion as assessed by ECG-gated PET in the healthy subjects was systematically underestimated compared with MRI. This underestimation correlated linearly with endocardial excursion during PET end diastole as measured by MRI. Myocardial end-diastolic wall thickness was systematically overestimated by PET, whereas end-systolic thickness deviated less than 1 mm from MRI. Cavity volumes measured by PET correlated linearly with MRI, with a tendency toward an underestimation of end-diastolic cavity volumes by PET. CONCLUSIONS: Absolute measures of cardiac structure and function may be obtained with a reasonable degree of accuracy by use of ECG-gated PET imaging. However, a high ECG-gating frequency appears to be required to obtain measurements comparable to what may be achieved by MRI. PMID- 14752473 TI - Discordance between exercise SPECT lung Tl-201 uptake and left ventricular transient ischemic dilation in patients with CAD. AB - BACKGROUND: In patients with coronary artery disease (CAD), the characteristics of those with discordant exercise thallium 201 single photon emission computed tomography (SPECT) lung uptake (lung-to-heart [L/H] ratio) and left ventricular (LV) transient ischemic dilation (LVTID) are not well defined. METHODS AND RESULTS: The population included 310 patients having exercise Tl-201 SPECT and coronary angiography. The population was subclassified into 4 subgroups: increased L/H ratio only, increased LVTID only, both, and neither. The L/H ratio was weakly correlated to LVTID (r = 0.18). The L/H ratio was correlated to the summed difference score (r = 0.26), summed rest score (r = 0.31), summed stress score (r = 0.5), and rest and stress LV volume (r = 0.5 and r = 0.54, respectively). LVTID was only correlated to the summed difference score (r = 0.32) and stress LV volume (r = 0.17). Increased LVTID only was associated with more frequent ischemia and patients with it tended to be more extensively ischemic, as compared with patients with increased L/H ratio only, but had a similar angiographic extent of CAD. These results were independent of prior myocardial infarction variable. CONCLUSIONS: As compared with patients with increased L/H ratio alone, patients with increased LVTID alone are more frequently ischemic but have a similar angiographic extent of CAD. Increased L/H ratio was correlated to both rest and postexercise LV volume, whereas increased LVTID was correlated only to postexercise LV volume. PMID- 14752474 TI - Advances in quantitative perfusion SPECT imaging. AB - Quantitative software for myocardial perfusion single photon emission computed tomography (SPECT) has advanced significantly over the last 25 years. The strength and availability of quantitative tools for perfusion SPECT have in many ways provided a competitive advantage to nuclear cardiology compared with other higher-resolution noninvasive imaging modalities for the detection of coronary artery disease. The purpose of this report is to review the advances in quantitative diagnostic software for cardiac SPECT over the past 25 years. The time period ending with the 1980s ("the past") saw the origins of nuclear cardiology with the development of planar thallium 201 imaging and perfusion SPECT imaging without electrocardiographic gating. The period from 1990 to the present saw the development of gated SPECT imaging providing both perfusion and functional information and attenuation correction SPECT with improved perfusion information. The report concludes with a look into the future, where hybrid multimodality imaging systems may provide a comprehensive noninvasive evaluation with previously unmatched accuracy in a single imaging session. PMID- 14752475 TI - Myocardial perfusion imaging agents: SPECT and PET. PMID- 14752476 TI - Emergency department assessment of patients with acute chest pain: myocardial perfusion imaging, blood tests, or both? PMID- 14752477 TI - The conundrum of left bundle branch block. PMID- 14752478 TI - Vasodilator stress myocardial perfusion examination discrepancy with coronary angiography: clarification with intravascular ultrasound. PMID- 14752479 TI - Acute myocardial infarction during adenosine myocardial perfusion imaging. PMID- 14752480 TI - Tl-201 reinjection enhances the detection of myocardial ischemia after acute myocardial infarction. PMID- 14752482 TI - Fixed-apex mitral annular descent correlates better with left ventricular systolic function than does free-apex left ventricular long-axis shortening. AB - Echocardiographic measures of mitral annular descent (MAD) assume a fixed left ventricular (LV) apex throughout the cardiac cycle, ignoring the apical component of LV long-axis shortening (LAS). We tested whether apical motion contributes significantly to LAS, making LAS a better surrogate of LV systolic function than MAD. Three-dimensional LV systolic MAD, LAS, and apical motion were measured in sheep using implanted radiopaque markers and biplane videofluoroscopy. End diastolic volume-stroke work relationship (preload recruitable stroke work) was computed as a load-independent index of LV systolic function. Apical motion was 1.4 +/- 0.8 mm, representing 22% of LAS (P <.05). Linear regression demonstrated that MAD correlated slightly better with preload recruitable stroke work (r = 0.808) than LAS (r = 0.792, both P <.001). Receiver operating characteristic curves demonstrated MAD was more accurate in predicting depressed LV function than LAS (93% vs 84%, respectively). Although LV apical motion contributed significantly to LAS, MAD measured with a fixed-apex assumption, as currently done echocardiographically, correlated more closely with LV preload recruitable stroke work. PMID- 14752483 TI - Simple detection of occluded coronary artery using retrograde flow in septal branch and left anterior descending coronary artery by transthoracic Doppler echocardiography at rest. AB - BACKGROUND: We hypothesized that coronary flow assessment by transthoracic Doppler echocardiography (TTDE) for both intramyocardial collateral channel and epicardial channels would be useful for identifying occluded left anterior descending coronary artery (LAD). METHODS: We assessed flow direction in the LAD and the septal branch (SEP) by TTDE in 302 consecutive patients who were suggested to have ischemic heart disease. We defined antegrade LAD flow as a direction from the base to the apex of the left ventricle in the anterior groove area, and antegrade SEP flow as a direction from anterior to inferior in the anterior interventricular septum. By contrast, we defined retrograde LAD and SEP flow as an inverse direction. We performed angiography on all patients. RESULTS: Retrograde flow was detected in 22 (LAD, 16 patients; SEP, 6 patients) of 23 patients with occluded LAD, and antegrade flow was detected in all patients without occluded LAD. The sensitivity and specificity for identification of occluded LAD by TTDE were 96% and 100%, respectively. CONCLUSIONS: Assessment of flow direction in both LAD and SEP by TTDE is a useful method in identification of occluded LAD. PMID- 14752484 TI - Pharmacologic stress echocardiography predicts total mortality early after acute myocardial infarction. AB - OBJECTIVE: The aim of this multicenter, prospective, observational study was to assess the value of inducible ischemia in a large population of survivors of a first uncomplicated myocardial infarction (MI). METHODS AND RESULTS: Pharmacologic stress echocardiography either with high-dose dipyridamole (0.84 mg/kg over 10 minutes) or high-dose dobutamine (up to 40 microg/kg over 3 minutes) (DET) was performed 9 +/- 10 days after a first acute uncomplicated MI in 1681 patients (1499 males; 57 +/- 10 years) with technically satisfactory rest echocardiographic study. Patients were followed up for a mean of 16 +/- 18 months (range: 1-122). DET was positive for myocardial ischemia in 884 (52.5%) and negative in 797 (47.5%) patients. During the follow-up there were 49 deaths for all-cause mortality (2.9% of the total population), 22 of which were cardiac; 62 (3.6%) nonfatal MIs; and 164 (9.7%) hospital readmissions for unstable angina. In all, 376 patients (22%) underwent coronary revascularization (bypass operation or angioplasty). RESULTS: Hard events occurred in 71 of the 884 patients with positive and in 40 of the 797 patients with negative DET (8% vs 5%, P =.014). Using the Cox proportional hazards model, age (relative risk [RR] 1.07, 95% confidence interval [CI] 1.03-1.1), history of angina (RR 3.8, 95% CI 1.6-8.6), peak wall-motion score index (RR 2.2, 95% CI 1.1-4.4), and pharmacologic dose at ischemia (RR 1.5, 95% CI 1.04-2.3) were independent predictors of all-cause death. CONCLUSIONS: In survivors of a first acute uncomplicated MI DET allows effective risk stratification on the basis of the presence, severity, and extent the induced ischemia. PMID- 14752485 TI - Relationship between the ischemic threshold at the onset of wall-motion abnormality on semisupine exercise echocardiography and the extent of coronary artery disease. AB - Semisupine exercise echocardiography (SSEE) provides the unique opportunity of continuous monitoring of segmental wall motion during physiologic stress. We evaluated the relationship between the ischemic threshold at the onset of wall motion abnormality on SSEE and the extent of coronary artery disease (CAD) in a consecutive series of 224 patients who underwent coronary angiography. Ischemic threshold was significantly lower for patients with multivessel disease compared with single-vessel disease: maximal workload was 102 versus 135 W (P = 1.3.10( 6)); percentage of maximal predicted heart rate achieved was 64 versus 70% (P =.004); and double product was 21,335 versus 23,389 (P =.03), respectively. Sensitivity, specificity, and positive and negative predictive values of SSEE for the detection of significant CAD (> or =60% diameter stenosis) were 81%, 74%, 90%, and 56%, respectively. SSEE is an accurate tool to diagnose CAD and the ischemic threshold at the onset of wall-motion abnormality is inversely related to the extent of CAD. PMID- 14752486 TI - Decreased right ventricular function after coronary artery bypass grafting and its relation to exercise capacity: a tricuspid annular motion-based study. AB - BACKGROUND: Decreased right ventricular (RV) function is a known echocardiographic finding after coronary artery bypass grafting (CABG). For patients with heart failure, RV dysfunction is a predictor of poor exercise capacity. The significance and time course of RV dysfunction and its relation to exercise capacity after CABG have not been elucidated, however. OBJECTIVES: In this prospective study, we assessed RV function measured from echocardiographic tricuspid annular motion (TAM) before and after CABG and its relation to exercise capacity. METHODS: In 99 patients accepted for CABG, we did a baseline echocardiographic investigation before operation, followed by repeated echocardiograms 3 months and 1 year after CABG. RV function was assessed using the magnitude of TAM measured at the RV free wall. An exercise stress test and coronary angiography were performed before and 3 months after CABG. RESULTS: RV function assessed by TAM was significantly reduced 3 months after CABG (22.4 vs 14.5 mm, P <.001) compared with preoperative measurements and remained so after 1 year (14.7 mm, P <.001). Left ventricular systolic function was unchanged 3 months after CABG. The 1-year echocardiographic follow-up showed paradoxical septal movement in 96% of the patients. Exercise capacity improved significantly 3 months after CABG compared with before (1.6 vs 1.83 W/kg, P <.001). These finding are independent of the state of the right coronary artery. CONCLUSIONS: One year after CABG, RV function remained depressed and septal motion remained paradoxical compared with the preoperative investigation, suggesting that these postoperative findings might be permanent in the majority of patients. Despite the reduced RV function, exercise performance 3 months after CABG was improved. The depressed RV function, measured from TAM after CABG, probably lacks clinical significance. PMID- 14752487 TI - Noninvasive quantification of regional myocardial function using Doppler-derived velocity, displacement, strain rate, and strain in healthy volunteers: effects of aging. AB - Quantification of regional function can be performed using Doppler tissue echocardiography to evaluate myocardial velocity, tissue displacement, strain, and strain rate. Although these techniques have been validated in animal experiments and early clinical trials, there are only limited measurements in healthy populations to use as reference data. In 100 healthy volunteers, left ventricular myocardial velocity, displacement, strain, and strain rate were measured using Doppler tissue echocardiography. Measurements were obtained from basal, mid, and apical segments of walls visualized from left ventricular apical 4- and 2-chamber views. Analysis of covariance was used to examine the effects of age and wall segment position. All parameters showed a strong dependence on wall segment position. Although myocardial velocities and strain rate showed significant dependence on age, displacement and peak systolic strain measures were less affected. Like pulsed Doppler mitral inflow velocity, tissue velocity and strain rate show age-related changes. PMID- 14752488 TI - Late left ventricular diastolic flow propagation velocity determined by color M mode Doppler in the assessment of diastolic dysfunction. AB - BACKGROUND: Early color M-mode Doppler flow propagation (Ep) through the left ventricle (LV) has been proposed as a useful noninvasive index for assessing LV relaxation, whereas data concerning late velocity propagation (Ap) is lacking. METHODS: We studied 51 patients with delayed relaxation (group I) and 50 with pseudonormal filling pattern (group II). Another 51 aged-matched healthy persons served as the control group. RESULTS: Patients showed increased left atrial dimensions, atrial wave of the pulmonary vein flow, and Ap, and reduced LV ejection fraction, Ep, and Ep/Ap ratio compared with the control group. Patients in group II revealed increased left atrial dimensions (P =.001), atrial wave of the pulmonary vein flow (P <.001), and Ep/Ap ratio (P <.001), and reduced LV ejection fraction and Ap (P <.001) compared with group I. Regression analysis showed that the strongest independent variable distinguishing normal from pseudonormal filling pattern was the Ep/Ap ratio. CONCLUSION: Ap evaluation offers a new diagnostic diastolic index, especially in the field of the pseudonormal pattern where the separation from normal is difficult. PMID- 14752489 TI - Assessment of regional left ventricular filling dynamics using color kinesis in patients with hypertrophic cardiomyopathy. AB - OBJECTIVES: Using color kinesis, we evaluated regional left ventricular filling dynamics in patients with hypertrophic cardiomyopathy (HCM). METHODS: In all, 20 patients (14 men and 6 women) and 20 age-matched healthy control subjects (10 men and 10 women) were studied. From color kinesis diastolic images in a left ventricular short-axis view, we generated regional time curves (6 segments) of left ventricular filling. The percent filling fraction at 25%, 50%, and 75% of filling time was averaged for all segments in each patient, and the SD of its mean was used as an asynchrony index at each particular filling time. The mean filling time for each segment was also measured. RESULTS: The asynchrony index was increased significantly in mid (50%) to late (75%) diastole in patients with HCM as compared with control subjects. Patients with HCM had regional mean filling times prolonged even in the nonhypertrophic segments. Moreover, there was significant correlation between the asynchrony index at mid and late diastole, and the global mean filling time. CONCLUSIONS: Color kinesis is useful in evaluating regional filling dynamics in patients with HCM. Our data reinforce the notion that HCM is a functionally heterogeneous disorder. PMID- 14752490 TI - Genesis of the restrictive filling pattern: pericardial constraint or myocardial restraint. AB - BACKGROUND: Restrictive filling pattern has been predictive of heart failure in patients with cardiomyopathy and after myocardial infarction, and is similar to the filling pattern in constrictive pericarditis and amyloid heart disease. The purpose of this study was to determine the role of both myocardial restraint and pericardial constraint in a chronic left ventricular dysfunction model with restrictive filling. METHODS: After instrumentation, a flat balloon containing a high-fidelity pressure catheter was inserted through a pericardial incision in 12 dogs with chronic left ventricular dysfunction. Intracardiac volume (ICV) was manipulated by inferior venal caval balloon occlusion and volume loading while hemodynamics, echo-assessed chamber size, and transmitral Doppler were obtained at the same atrial paced rate with an intact pericardium and after pericardiectomy. RESULTS: With an intact pericardium, deceleration time increased with reduced ICV (130 +/- 35 vs 153 +/- 47 milliseconds, P <.05) and shortened with increased ICV (107 +/- 45 milliseconds, P <.05). The filling fraction at one third of diastole decreased with reduced ICV (45.6 +/- 29.3 vs 24.2 +/- 15.8%, P <.01) and increased with increased ICV (60.1 +/- 14.8%, P <.05). Deceleration time could be predicted from intrapericardial pressure, the transmural left ventricular chamber stiffness constant, and filling fraction at one-third of diastole. After pericardiectomy, deceleration time also shortened with increased ICV (141 +/- 26 vs 112 +/- 38 milliseconds, P <.01). However, filling fraction at one-third of diastole was markedly reduced at paced baseline (19.9 +/- 14.4%, P <.01) and with increased ICV (15.5 +/- 11.8%, P <.001) as compared with an intact pericardium. CONCLUSIONS: Pericardial constraint and myocardial restraint play a role in restrictive filling pattern. Pericardial constraint becomes evident with redistribution of diastolic filling to later in diastole after pericardiectomy. PMID- 14752491 TI - Baseline echocardiographic values for adult male rats. AB - BACKGROUND: Because of safety, repeatability, and portability, clinical echocardiography is well established as a standard for cardiac anatomy, cardiac function, and hemodynamics. Similarly, application of echocardiography in commonly used rat experimental models would be worthwhile. The use of noninvasive ultrasound imaging in the rat is a potential replacement for more invasive terminal techniques. Although echocardiography has become commonly used in the rat, normal parameters for cardiac anatomy and function, and comparison with established human values, have not been reported. METHODS: A total of 44 Sprague Dawley male rats had baseline echocardiography replicating a protocol for clinical echocardiography. RESULTS: Complete 2-dimensional echocardiography for cardiac anatomy and function was obtained in 44 rats. Hemodynamic parameters could be recorded in 85% of rats. The ejection fraction and fractional shortening values of the left ventricle were similar to those reported for healthy human beings. Pulsed Doppler velocities of atrial systole for mitral valve inflow, pulmonary vein reversal, and Doppler tissue of the lateral mitral valve annulus also had similar means as healthy human beings. The calculated left ventricular mass was at the same order of magnitude as a proportion of body weight of rat to man. All other observations in the clinical protocol were different from those reported in healthy human beings. CONCLUSION: The use of echocardiography for assessment of cardiac anatomy, function, and hemodynamics can be consistently applied to the rat and replicates much of the information used routinely in human echocardiography. PMID- 14752492 TI - Should we be evaluating the ventricle or the myocardium? Advances in tissue characterization. PMID- 14752493 TI - Acute myocardial infarction caused by embolism of thrombus in the right coronary sinus of Valsalva: a case report and review of the literature. AB - A 56-year-old man presented with sustained chest pain. Coronary angiography revealed total occlusion of the distal right coronary artery and left anterior descending branch. Left ventriculography depicted a mobile mass in the right sinus of Valsalva originating from the ostium of the right coronary artery. Transesophageal echocardiography (TEE) showed a mobile mass in the sinus of Valsalva and another mobile mass in the aortic arch. The mass at the right sinus of Valsalva was surgically resected, and histologic examination revealed an organized thrombus. Coagulation study showed protein S deficiency. This is the first case of acute myocardial infarction as a result of multiple coronary embolism caused by thrombosis in the right sinus of Valsalva with a second aortic arch thrombosis, contributed by protein S deficiency. PMID- 14752494 TI - Demonstration of coronary artery flow using transthoracic Doppler echocardiography. AB - Transthoracic Doppler echocardiography is emerging as a promising method for evaluating coronary artery disease. After a period of training, detection and measurement of distal left anterior descending coronary artery flow with transthoracic Doppler echocardiography is feasible in more than 90% of the patients. Using transthoracic Doppler echocardiography with a high-frequency transducer and special setting of low Nyquist limits, pathologic coronary flow dynamics can be demonstrated. Measurement of coronary flow reserve may impact diagnosis or clinical treatment in those: (1) with anginal chest pain and angiographically normal coronary arteries; (2) with intermediate-grade coronary obstruction where the physiologic significance is in doubt; and (3) who have had an attempt at revascularization and the effectiveness of the therapy is uncertain. PMID- 14752495 TI - Right ventricular hemangioma causing right ventricular inflow obstruction and right heart failure. AB - We report a patient who presented with symptoms of right heart failure. Transesophageal echocardiography revealed a right ventricular mass, causing right ventricular inflow obstruction. Coronary angiography revealed a characteristic tumor blush. After successful surgical resection, histologic findings were consistent with a right ventricular hemangioma. PMID- 14752496 TI - Fracture of the connecting bar of a stent graft in the thoracic aorta: a diagnosis by echocardiography. PMID- 14752497 TI - Hepatocellular carcinoma with invasion into right cardiac cavities: report of a case and literature review. AB - Hepatocellular carcinoma is a tumor that rarely invades the inferior vena cava and right heart. We describe the case of a 63-year-old man with jaundice and a history of cryptogenic hepatic cirrhosis with diagnosis of hepatocellular carcinoma. He was referred to the echocardiography laboratory for sudden dyspnea and suggestion of pulmonary thromboembolism. Echocardiographic study reported a mass in the right atrium proceeding from the inferior vena cava. PMID- 14752498 TI - Noninvasive tests for arterial structure, function, and compliance: do they identify risk or diagnose disease? AB - BACKGROUND: Brachial artery reactivity (BAR), carotid intima-media thickness (IMT), and applanation tonometry for evaluation of total arterial compliance may provide information about preclinical vascular disease. We sought to determine whether these tests could be used to identify patients with coronary artery disease (CAD) without being influenced by their ability to identify those at risk for CAD developing. METHODS: We studied 100 patients and compared 3 groups: 35 patients with known CAD; 34 patients with symptoms and risk factors but no CAD identified by stress echocardiography (risk group); and 31 control subjects. BAR and IMT were measured using standard methods, and total arterial compliance was calculated by the pulse-pressure method from simultaneous radial applanation tonometry and pulsed wave Doppler of the left ventricular outflow. Ischemia was identified as a new or worsening wall-motion abnormality induced by stress. RESULTS: In a comparison between the control subjects and patients either at risk for developing CAD or with CAD, the predictors of risk for CAD were: age (P =.01); smoking history (P =.002); hypercholesterolemia (P =.002); and hypertension (P =.004) (model R = 0.82; P =.0001). The independent predictors of CAD were: IMT (P =.001); BAR (P =.04); sex (P =.005); and hypertension (P =.005) (model R = 0.80; P =.0001). CONCLUSION: IMT, BAR, and traditional cardiovascular risk factors appear to identify patients at risk for CAD developing. However, only IMT was significantly different between patients at risk for developing CAD and those with overt CAD. PMID- 14752500 TI - A case of familial trichomegaly in association with oculocutaneous albinism type 1. PMID- 14752501 TI - Posterior polymorphous dystrophy with polycoria and corectopia. PMID- 14752502 TI - Outcome of ICG-assisted ILM peel in macular hole surgery. AB - PURPOSE: To investigate whether indocyanine green (ICG) staining of the internal limiting membrane (ILM) improves surgical outcome. METHODS: A total of 34 patients were identified who underwent macular hole surgery with ILM peeling augmented with ICG. These were matched retrospectively with 34 patients who underwent macular hole surgery without the use of ICG. Closure rates from primary and secondary procedures were compared, as were changes in Snellen and Logmar visual acuity. RESULTS: There was no significant difference between the two groups with reference to demographic features of age, sex, staging of the macular holes, and the proportion subsequently undergoing cataract surgery. The mean follow-up period was 7.7 months in the ICG group and 6.3 months in the non-ICG group. Closure rates from primary surgery were 91.2% in the ICG group and 73.5% in the non-ICG group (P = 0.056), whereas overall closure rates following further surgery were similar in both groups (94.1 and 91.2%, respectively). Logmar visual improvement above baseline was achieved in 53% of the non-ICG group compared to 82% of the ICG group (P = 0.01). Where primary hole closure occurred, there was no statistically significant difference in visual outcome between the two groups. CONCLUSION: ICG-assisted ILM peel in macular hole surgery is associated with a higher closure rate following a single surgical procedure. This difference was found to approach statistical significance (P = 0.056). Overall visual improvement was greater in the ICG group (P = 0.01); however, this reflected the higher closure rates. Visual outcomes between the two groups were comparable where primary hole closure occurred. PMID- 14752503 TI - Ultrasound biomicroscopic documentation of traumatic cyclodialysis cleft closure with hypotony by medical therapy. PMID- 14752504 TI - Endogenous endophthalmitis secondary to Lemierre's syndrome. PMID- 14752505 TI - Longitudinal study examining the risk factors for proliferative retinopathy and maculopathy in type-I diabetes: The Royal College of Physicians of Edinburgh Diabetes Register Group. AB - PURPOSE: The aim of this study was to determine whether there were any differences in the risk factors for developing proliferative diabetic retinopathy or maculopathy in patients with type-I diabetes. METHOD: In all, 1632 patients aged 35 years or younger at diagnosis and treated with insulin, attending six hospital diabetes clinics in Scotland and included on the Royal College of Physicians of Edingburgh Diabetes Register were followed up for a median of 4.0 (2.5-5.5 years: interquartile range). All patients were screened at least annually for diabetic retinopathy using direct ophthalmoscopy, and positive findings were confirmed using slit lamp by an ophthalmologist. RESULTS: Duration of diabetes and HbA1c were the important risk factors for developing proliferative retinopathy, while the duration of diabetes, systolic blood pressure, and HbA1c were the important factors of maculopathy. The adjusted relative incidence for proliferative retinopathy with a HbA1c in the highest quartile was 26.7, while for maculopathy it was only 2.29. Carstairs deprivation score was not associated with either retinal pathology. There was a plateau effect for systolic blood pressure of 140 mmHg and for duration of diabetes of 16 years for developing either maculopathy or proliferative retinopathy. CONCLUSION: Duration of diabetes is a strong predictor for maculopathy and proliferative disease, but is relatively more important for proliferative disease. Raised systolic blood pressure is relatively more important for predicting maculopathy, while raised HbA1c is relatively more important for developing proliferative retinopathy. PMID- 14752506 TI - Progression of visual field defect in a normal-tension glaucoma patient after laser in situ keratomileusis. PMID- 14752507 TI - Characteristics and surgical outcomes of paediatric retinal detachment. AB - PURPOSE: To report the clinical features and surgical and visual outcomes of rhegmatogenous retinal detachment (RRD) in the paediatric population. METHODS: A retrospective review of children (aged 0-15 years) who underwent primary surgical repair for RRD at the Hiroshima University Hospital between 1988 and 2001. RESULTS: In all 53 eyes of 49 patients were identified; paediatric RRD accounted for 3.1% of 1779 eyes with RRD operated on during this period. The causes of RRD included blunt trauma (27%), myopia (25%), idiopathic (20%), familial exudative vitreoretinopathy (13%), and others. Among 55 eyes, 12 (22%) already had proliferative vitreoretinopathy (PVR) of grade C or D preoperatively. The median initial visual acuity (VA) was 0.3. Retinal reattachment was achieved with a single operation in 78%. Final retinal reattachment was achieved in 87%. Retinal reattachment rates with and without PVR were 42% and 100%, respectively (P<0.01). Median final VA was 0.7. Final VA was > or =0.1 in 73% and > or =0.5 in 53%; four eyes had a final VA of no light perception. The presence of preoperative PVR (P=0.03) and the initial VA (P<0.0001) significantly affected final VA. CONCLUSIONS: Paediatric RRD is characterised by a delay in diagnosis, as evidenced by the high rate of PVR at presentation. Retinal reattachment was adversely affected by the presence of PVR. Final VA correlated with the initial VA and was significantly affected by preoperative PVR. Early diagnosis may improve the visual prognosis of paediatric retinal detachment. PMID- 14752508 TI - Ischemia-induced neuronal cell death is mediated by the endoplasmic reticulum stress pathway involving CHOP. AB - Brain ischemia induces apoptosis in neuronal cells, but the mechanism is not well understood. When wild-type mice were subjected to bilateral common carotid arteries occlusion (BCCAO) for 15 min, apoptosis-associated morphological changes and appearance of TUNEL-positive cells were observed in the striatum and in the hippocampus at 48 h after occlusion. RT-PCR analysis revealed that mRNAs for ER stress-associated proapoptotic factor CHOP and an ER chaperone BiP are markedly induced at 12 h after BCCAO. Immunohistochemical analysis showed that CHOP protein is induced in nuclei of damaged neurons at 24 h after occlusion. In contrast, ischemia-associated apoptotic loss of neurons was decreased in CHOP(-/ ) mice. Primary hippocampal neurons from CHOP(-/-) mice were more resistant to hypoxia-reoxygenation-induced apoptosis than those from wild-type animals. These results indicate that ischemia-induced neuronal cell death is mediated by the ER stress pathway involving CHOP induction. PMID- 14752509 TI - Radiation-induced apoptosis in developing mouse retina exhibits dose-dependent requirement for ATM phosphorylation of p53. AB - Ionizing radiation (IR) induces DNA breakage to activate cell cycle checkpoints, DNA repair, premature senescence or cell death. A master regulator of cellular responses to IR is the ATM kinase, which phosphorylates a number of downstream effectors, including p53, to inhibit cell cycle progression or to induce apoptosis. ATM phosphorylates p53 directly at Ser15 (Ser18 of mouse p53) and indirectly through other kinases. In this study, we examined the role of ATM and p53 Ser18 phosphorylation in IR-induced retinal apoptosis of neonatal mice. Whole body irradiation with 2 Gy IR induces apoptosis of postmitotic and proliferating cells in the neonatal retinas. This apoptotic response requires ATM, exhibits p53 haploid insufficiency and is defective in mice with the p53S18A allele. At a higher dose of 14 Gy, retinal apoptosis still requires ATM and p53 but can proceed without Ser18 phosphorylation. These results suggest that ATM activates the apoptotic function of p53 in vivo through alternative pathways depending on IR dose. PMID- 14752510 TI - hsp70-DnaJ chaperone pair prevents nitric oxide- and CHOP-induced apoptosis by inhibiting translocation of Bax to mitochondria. AB - We reported that the endoplasmic reticulum (ER) stress pathway involving CHOP, a member of the C/EBP transcription factor family, plays a key role in nitric oxide (NO)-mediated apoptosis of macrophages and pancreatic beta cells. We also showed that the cytosolic chaperone pair of hsp70 and dj1 (hsp40/hdj-1) or dj2 (HSDJ/hdj 2) prevents NO-mediated apoptosis upstream of cytochrome c release from mitochondria. To analyze roles of the chaperone pair in preventing apoptosis, RAW 264.7 macrophages stably expressing hsp70 and dj1 or dj2 were established. The chaperone pair prevented LPS/IFN-gamma-induced and NO-mediated apoptosis downstream of CHOP induction. hsp70 mutant protein lacking the ATPase domain or the C-terminal EEVD sequence were not effective in preventing CHOP-induced apoptosis. A mutant dj2 lacking the C-terminal prenylation CaaX motif, was also not effective. When wild-type RAW 264.7 cells were treated with LPS/IFN-gamma, NO mediated apoptosis was induced, and proapoptotic Bcl-2 family protein Bax was translocated from cytosol to mitochondria. This translocation was prevented in cells stably expressing hsp70/dj2, and in CHOP knockout cells. Overexpression of CHOP in wild-type cells also induced translocation of Bax and this translocation was prevented in cells expressing hsp70/dj2. CHOP-induced apoptosis was prevented by Bax knock-down. Coimmunoprecipitation experiments showed that Bax interacts with both hsp70 and dj1/dj2. ATPase domain of hsp70 was necessary for the binding with Bax. These findings indicate that CHOP-induced apoptosis is mediated by translocation of Bax from the cytosol to the mitochondria, and hsp70/dj1 or dj2 chaperone pair prevents apoptosis by interacting with Bax and preventing translocation to the mitochondria. PMID- 14752511 TI - Transcription factor AP-2alpha triggers apoptosis in cardiac myocytes. AB - Idiopathic-dilated cardiomyopathy (IDC) is a common primary myocardial disease of unknown etiology associated with apoptosis, cardiac dilatation, progressive heart failure and increased mortality. An elevation of the transcription factor activator protein 2alpha (AP-2alpha) is involved in vertebrate embryonic development and oncogenesis. Here, we show that AP-2alpha protein is expressed in the human heart and increased in human failing myocardium with IDC. Adenovirus mediated overexpression of human AP-2alpha triggered apoptosis and increased mRNA levels of Bcl-2 family members Bax and Bcl-x in rat cardiomyocytes. Immunohistological analysis of human myocardium revealed an increased percentage of AP-2alpha-positive nuclei in IDC and, interestingly, a colocalization of AP 2alpha-positive but not -negative cells with a caspase-cleaved fragment of poly(ADP-ribose)polymerase. We suggest AP-2alpha as a novel cardiac regulator implicated in the activation of apoptosis in IDC. PMID- 14752512 TI - Human alphaA- and alphaB-crystallins bind to Bax and Bcl-X(S) to sequester their translocation during staurosporine-induced apoptosis. AB - AlphaA- and alphaB-crystallins are distinct antiapoptotic regulators. Regarding the antiapoptotic mechanisms, we have recently demonstrated that alphaB crystallin interacts with the procaspase-3 and partially processed procaspase-3 to repress caspase-3 activation. Here, we demonstrate that human alphaA- and alphaB-crystallins prevent staurosporine-induced apoptosis through interactions with members of the Bcl-2 family. Using GST pulldown assays and coimmunoprecipitations, we demonstrated that alpha-crystallins bind to Bax and Bcl-X(S) both in vitro and in vivo. Human alphaA- and alphaB-crystallins display similar affinity to both proapoptotic regulators, and so are true with their antiapoptotic ability tested in human lens epithelial cells, human retina pigment epithelial cells (ARPE-19) and rat embryonic myocardium cells (H9c2) under treatment of staurosporine, etoposide or sorbitol. Two prominent mutants, R116C in alphaA-crystallin and R120G, in alphaB-crystallin display much weaker affinity to Bax and Bcl-X(S). Through the interaction, alpha-crystallins prevent the translocation of Bax and Bcl-X(S) from cytosol into mitochondria during staurosporine-induced apoptosis. As a result, alpha-crystallins preserve the integrity of mitochondria, restrict release of cytochrome c, repress activation of caspase-3 and block degradation of PARP. Thus, our results demonstrate a novel antiapoptotic mechanism for alpha-crystallins. PMID- 14752513 TI - Targeting Bcl-xL in esophageal squamous cancer to sensitize to chemotherapy plus TRAIL-induced apoptosis while normal epithelial cells are protected by blockade of caspase 9. PMID- 14752516 TI - Take a cold flower. PMID- 14752517 TI - Monitoring p53's pulse. PMID- 14752518 TI - Close look at gene conversion hot spots. PMID- 14752519 TI - Getting our AKT together in schizophrenia? PMID- 14752520 TI - RASSF1A, the new guardian of mitosis. PMID- 14752523 TI - Analysis of HER-2/neu amplification in endometrial carcinoma by chromogenic in situ hybridization. Correlation with fluorescence in situ hybridization, HER 2/neu, p53 and Ki-67 protein expression, and outcome. AB - Fluorescence in situ hybridization (FISH) is the most widely used technique to detect HER-2/neu gene amplification; however, it is only available in some institutions. In contrast, chromogenic in situ hybridization (CISH) can be evaluated by routine light microscopy. In endometrial carcinoma there are few data concerning HER-2/neu status and prognosis. Therefore, we determined HER 2/neu gene status by CISH using a digoxigenin-labelled probe on 60 formalin-fixed paraffin-embedded endometrial carcinomas. The data were compared with the immunohistochemistry of HER-2/neu (A0485, TAB250), p53, Ki-67, clinicopathological factors, and survival. By conventional light microscopy, HER 2/neu amplification (>/=6 copies >50% cancer cells) was detected in 14% (8/59) tumours, HER-2/neu overexpression (>10% cells moderate/strong complete membrane staining) in 22% (13/60) for A0485, and 18% (11/60) for TAB250, p53 (>10% +cells) in 61% (36/59), and Ki-67 (>50% +cells) in 50% (30/60). Discordant cases for CISH and immunohistochemistry, as well as all (2+) were further analysed by FISH (Vysis). Among 10 cases (2+) and not amplified by CISH, two showed low-level amplification by FISH. Significant correlation was found between amplification and protein overexpression (P5 dollars per week, and smoking in the house. CONCLUSIONS: Parental behaviour is a key determinant of smoking by New Zealand adolescents. Efforts that target the role of parents should be pursued, such as health promotion strategies that advise parents about the possible benefits of banning smoking in the home, limiting pocket money, and not providing cigarettes to their children. PMID- 14752536 TI - Digoxin therapeutic drug monitoring: an audit and review. AB - AIM: The measurement and assessment of digoxin concentrations are often performed poorly. We have conducted an audit to assess the appropriateness of digoxin therapeutic drug monitoring in Christchurch Hospital. METHODS: One hundred consecutive requests for digoxin concentrations in Christchurch Hospital inpatients were assessed. The case notes and hospital medication records were reviewed to determine the indication for testing, the appropriateness of the sampling time and of the subsequent alteration to dosing. RESULTS: In 53% of requests no clear indication for digoxin therapeutic drug monitoring (TDM) could be determined. In the remainder, 'suspected toxicity' accounted for 31% and 'therapeutic failure' for 16%. Samples were inappropriately taken within eight hours post-dose in 32% of requests. In 19% of cases, the samples did not reflect steady-state conditions. In 5% of occasions, the subsequent decision regarding dose adjustment was felt to be clearly inappropriate, and there was uncertainty regarding appropriateness in some other cases. Overall, in only 29% of requests was TDM performed appropriately with regard to indication, sampling and subsequent dose alteration. CONCLUSIONS: At Christchurch Hospital, the practice of TDM for digoxin is often inappropriate. It would seem that medical staff education is required to improve this practice. PMID- 14752537 TI - The use of inhaled and related respiratory medications in Christchurch rest homes. AB - AIM: To describe the use of inhaled and related respiratory medications ('asthma medications') and associated management amongst Christchurch rest-home residents. METHODS: Fifty per cent of Christchurch rest homes were randomly selected. All residents on asthma medications, the rest-home managers, care-giving staff, and the residents' general practitioners were interviewed using specific questionnaires. RESULTS: All of the rest homes, residents using asthma medications and senior staff members participated. Seventy five per cent of caregivers and 73% of general practitioners took part. Asthma medications were used by 13% of 1416 rest-home residents. Eighty four per cent of these used a preventer medication, mostly inhaled steroids. Some daily doses exceeded current treatment guidelines. One third of residents using inhalers had an inadequate technique. Some staff and residents chose the wrong inhaler to manage 'shortness of breath'. Regular bronchodilator dosing, rather than 'as required', was common. Those using a spacer device usually had a good technique. Residents appreciated non-pharmacological strategies for breathlessness. Staff identified a need for clear written management plans. CONCLUSIONS: There were significant deficiencies in the staff and residents' knowledge of obstructive airways management and medications. Regular review of inhaler technique, greater use of spacers, and regular staff education may improve residents' respiratory management. Inhaled corticosteroids may be used in too high a dose. Inconsistent management of acutely deteriorating asthma/chronic obstructive pulmonary disease may be addressed by greater use of written management plans in residents' notes. PMID- 14752538 TI - Buteyko Breathing Technique for asthma: an effective intervention. AB - AIM: To assess the impact of the Buteyko Breathing Technique (BBT) on medication use in asthma. METHODS: A blinded randomised controlled trial comparing BBT with control was conducted in 38 people with asthma aged between 18 and 70. Participants were followed for six months following the intervention. Medication use and indices of ventilatory function were recorded. RESULTS: No significant change in FEV1 (forced expiratory volume in one second) was recorded in either group. The BBT group exhibited a reduction in inhaled steroid use of 50% and beta2-agonist use of 85% at six months from baseline. In the control group inhaled steroid use was unchanged and beta2-agonist use was reduced by 37% from baseline. Investigator contact between the two groups was equal. There were no adverse events recorded in either group. CONCLUSIONS: BBT is a safe and efficacious asthma management technique. BBT has clinical and potential pharmaco economic benefits that merit further study. PMID- 14752539 TI - Is the provision of laboratory results via the Internet acceptable to patients? A survey of private patients in a large, specialist gynaecology practice. AB - AIM: The purpose of this research project was to ascertain patients' views as to the acceptability of having their laboratory results communicated via the Internet. A database was designed to hold laboratory results on a web site. METHODS: A survey was then prepared for patients of a private, specialist gynaecology practice, at whom the site was targeted. RESULTS: Two hundred and thirty five patients responded to the questionnaire, which canvassed their views on the provision of results through this new modality. Of the 235 respondents, 190 professed to be users of the Internet in general. Of this group almost 60% used the laboratory results web site. Feedback was highly favourable with 87% of those who used the site stating that they would use it again in the future. The respondents as a whole rated the Internet as their preferred method for gaining their laboratory results, even if they had not used it in this instance. The number of patients who held concerns about the security of the site, or the PIN system used to access it, was significantly small. Neither of these possible problems seemed to be a barrier to patients' utilisation of the site. CONCLUSIONS: The Internet-based laboratory results were well received by those who accessed them and the figures for those who would use the system again in the future suggest that once converted patients were more than likely to use the site again. With the increasing use of the Internet for the dissemination of all sorts of information, laboratory results hosted in this way can only become more and more favourably accepted by patients. PMID- 14752540 TI - A case of primary amoebic meningoencephalitis: North Island, New Zealand. PMID- 14752541 TI - The Green Prescription: a field of dreams? AB - Increasing the physical activity of New Zealanders has assumed a higher priority with new primary healthcare strategies. Physical Educators, the professionals who research, teach and practise in the domain of physical activity and exercise, have raised concerns regarding the strategies employed. This essay details several misgivings with public health initiatives designed to increase physical activity, and makes a case for recognising the expertise of graduates specifically trained in this specialist field. A system of referral to accredited exercise professionals is suggested to be a more efficacious means of changing physical activity behaviours. PMID- 14752542 TI - Spontaneous haemopneumothorax: will it ever be common enough to raise some clinical suspicion? PMID- 14752543 TI - Calcium scores are the key to predicting cardiovascular risk. PMID- 14752544 TI - Towards a partnership of care for patients with haemophilia. PMID- 14752545 TI - Why is the NZMJ password protected? PMID- 14752546 TI - Time for the New Zealand cancer control strategy. PMID- 14752547 TI - A healthy paradigm shift in the analytical science community. PMID- 14752548 TI - Bioanalytical challenges for analytical chemists. AB - Analytical chemistry contributes significantly to the life sciences through developing measurement techniques that provide quantitative chemical information. Developments in biological research present new challenges for analytical chemistry and call for revolutionary methods and new thinking. Dana Spence outlines here some key and unique challenges encountered when making measurements on samples of a biological nature, supported with real examples from his own research. PMID- 14752549 TI - Droplets formation from microfluidic device for sampling and measurement of atmospheric nitrogen dioxide. AB - A new NO(2) measurement collection device developed in this study indicates efficient absorption, enough to be applied to the practical determination of atmospheric NO(2), which should become a useful tool compared with conventional methods. PMID- 14752550 TI - In vivo study of triazine herbicides in plants by SPME. AB - The use of SPME for in vivo monitoring of herbicide levels in plant tissues is evaluated. Fibers are exposed to the plant tissue with the aid of buffer located at the fiber/tissue interface region. Following this extraction period the extracted amount is estimated by solvent desorption and LC-MS-MS. PMID- 14752551 TI - Capillary isoelectric focusing in pseudo-closed channel coupled to matrix assisted laser desorption/ionization mass spectrometry for protein analysis. AB - Capillary isoelectric focusing (CIEF) was performed in pseudo-closed channel to separate proteins on a plastic chip. Pseudo-closed channel provided a novel way to couple protein separation by CIEF to MALDI mass spectrometry without eluting the focused proteins. PMID- 14752552 TI - BSE control: detection of gelatine-derived peptides in animal feed by mass spectrometry. AB - The epidemic of bovine spongiform encephalopathy (BSE) is thought to have resulted from feeding scrapie-infected sheep to cattle. This has led to a ban of feeding animals with "processed animal protein"(PAP). We report a novel approach for the mass spectrometric detection of PAP contamination in animal feedstuffs by detecting gelatine, a derivative of the major animal protein collagen. A method was developed to hydrolyse gelatine standards with hydrochloric acid, followed by detection of the derived hydrolytic peptides at m/z 828, 915, 957 and 1044 by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF MS) and liquid chromatography electrospray ionisation mass spectrometry (LC-ESI-MS-MS). The marker peptides can be detected at concentrations of 100 ng ml(-1) gelatine in water with MALDI. The procedure was adapted to obtain a suitable peptide map of gelatine extracted from spiked feed. The ratio of signal area of the gelatine-derived peptide at m/z 1044 to the internal standard at m/z 556 is shown to relate to the total amount of gelatine present in the sample. PMID- 14752553 TI - Preliminary studies on selenium-containing proteins in Brassica juncea by size exclusion chromatography and fast protein liquid chromatography coupled to ICP MS. AB - An approach for screening and resolving selenium-containing plant proteins was developed based on the combination of sample preparation and multi-dimensional liquid chromatography coupled to ICP-MS. Different protein extraction protocols were investigated. A 24 h dodecylsulfate-mediated protein extraction in a sonication bath followed by acetone precipitation was found to be optimal. The use of different protein precipitate solubilizing agents (sodium dodecyl sulfate media and Tris-HCl buffer) demonstrates possible fractionation of the selenium containing proteins. Selenium-containing protein screening and fractionation were carried out by means of SEC-ICP-MS. High molecular weight selenium containing proteins were solubilized with a sodium dodecyl sulfate-containing buffer, whereas the low molecular weight compounds were released into a Tris-HCl buffer. Size exclusion chromatography-fast protein liquid chromatography coupled to ICP MS allowed separation and detection of several selenium-containing proteins in Se supplemented wild type Brassica juncea plant, a fast growing selenium accumulator. PMID- 14752555 TI - Establishing the synthetic origin of amphetamines by 2H NMR spectroscopy. AB - Nine samples of N-acetyl-3,4-methylenedioxyamphetamine (N-acetyl-MDA), prepared according to the most common synthetic procedures, are submitted to (2)H NMR spectroscopy. The relative deuterium content at the various sites of the molecule is shown to depend on its synthetic history. The technique provides a chemical fingerprint of N-acetyl-MDAs and it can be used to trace back the precursor materials and the synthetic pathways employed in the preparation of the samples. PMID- 14752556 TI - Raman spectroscopic analysis of dragon's blood resins-basis for distinguishing between Dracaena(Convallariaceae), Daemonorops(Palmae) and Croton(Euphorbiaceae). AB - "Dragon[prime or minute]s blood" is the name applied to the deep-red coloured resin obtained from various plants. The original source in Roman times, used by many cultures and esteemed for its depth of colour and mystical association, was the dragon tree Dracaena cinnabari(Convallariaceae), found only on the Indian Ocean island of Socotra, (Yemen). Additional sources emerged later, including another species of Dracaena, D. draco, from the Canary Islands and Madeira, and species in the genera Daemonorops(Palmae) from South East Asia and Croton(Euphorbiaceae) from tropical parts of both the New and Old Worlds. In this study, examples of dragon's blood resins from the Economic Botany Collections at the Royal Botanic Gardens, Kew, dating from 1851 to 1993, have been analysed non destructively using Raman spectroscopy. The Raman spectra of well-documented, provenanced specimens have been used to establish the source of specimens of questionable or unknown origin. It has also been possible from the Raman spectra to indicate whether processing of the resins has been undertaken in the preparation of the specimens before their deposition at Kew. PMID- 14752557 TI - Electrospray ionization-ion mobility spectrometry: a rapid analytical method for aqueous nitrate and nitrite analysis. AB - This paper reports the first example of electrospray ionization (ESI) for the separation and detection of anions in aqueous solutions by ion mobility spectrometry (IMS). Standard solutions of arsenate, phosphate, sulfate, nitrate, nitrite, chloride, formate, and acetate were analyzed using ESI-IMS and distinct peak patterns and reduced mobility constants (K(0)) were observed for respective anions. Real world water samples were analyzed for nitrate and nitrite to determine the feasibility of using ESI-IMS as a rapid analytical method for monitoring nitrate and nitrite in water systems. The data showed satisfactory correlation between the measured value ([similar]0.16 ppm) and the reported maximum nitrate-nitrogen concentration (0.2 ppm) found in a local drinking water system. For on-site measurement applications, direct sample introduction and air as an alternate drift gas to nitrogen were evaluated. The identities of the nitrite and nitrate mobility peaks were verified by comparison of reduced mobility constants with mass identified nitrate and nitrite ions reported in literature. In the mixing ratio, a linear dynamic range of 3 orders of magnitude and instrument detection limits of 10 ppb for nitrate and 40 ppb for nitrite were obtained. The calibration curves showed r(2) value of 0.98 and slope of 0.06 for nitrate and r(2) value of 0.99 and slope of 0.11 for nitrite. PMID- 14752558 TI - Validation of a simple, sensitive method for the determination of beta-estradiol in bovine urine using gas-chromatography negative-ion chemical ionization mass spectrometry. AB - A new method is presented for the analysis of 17[small beta]-estradiol in bovine urine. After deconjugation, the sample is cleaned up using an OASIS[trade mark sign] HLB disposable cartridge and extracted into 1-chlorobutane. The hormone is derivatized using pentafluorobenzoyl chloride. The derivatized estradiol is quantitated using gas-chromatography negative-ion chemical ionization mass spectrometry. Calibrations, obtained using spiked blank urine, are linear in the range of 100-1000 pg mL(-1) with CC[small alpha] approximately 170 pg mL(-1) and CC[small beta] of 287 pg mL(-1). Recoveries are in the range of 80 to 130%. The method is rugged, rapid and sensitive when compared to other hormone methods. PMID- 14752559 TI - Simultaneous analysis of hepatotoxic pyrrolizidine alkaloids and N-oxides in comfrey root by LC-ion trap mass spectrometry. AB - The purpose of the current study was to develop a LC-MS(n) method for the analysis of pyrrolizidine alkaloids (PAs) in comfrey. Published data presents an extensive list of PAs and their N-oxides present in comfrey. However, standards are not commercially available for any of the PAs typically present in comfrey. Those PAs that are not stereoisomers were readily resolved on a C(18) column using a water-acetonitrile gradient as the mobile phase. The use of a selective technique, LC-MS/MS, allowed us to identify groups of PAs and their N-oxides, as well as identify the number of PAs present in each group, including those that were not completely resolved chromatographically. PMID- 14752561 TI - Protein microarray scanning in label-free format by Kelvin nanoprobe. AB - Surface-immobilized protein species deposited in the microarray format have been detected by time-of-flight secondary ion mass spectrometry and by scanning Kelvin nanoprobe. The former method was used to examine the nature of protein deposition on amine-coated glass slides and gold substrates in preparation for Kelvin measurements. Both gallium and SF(5)(+) ion sources were employed to produce positive and negative ion spectra of amino acids and polypeptides. Scanning Kelvin technology has been used to detect antibody-antigen interactions in a label-free protocol through measurement of the surface potential of the biochemical pair on indium tin oxide, amine-treated slides and gold substrates. The results show that good inter-spot reproducibility can be achieved and that deposited areas can be examined for homogeneity at 100 nm resolution. This work represents the first report on surface potential detection in protein microarray technology. PMID- 14752562 TI - Development of a rapid screening test for veterinary sedatives and the beta blocker carazolol in porcine kidney by ELISA. AB - Sedatives and tranquillisers are frequently used to reduce stress during the transportation of food producing animals. The most widely used classes of sedatives include the butyrophenone azaperone, the phenothiazines acepromazine, propionylpromazine, chlorpromazine and the [small beta]-blocker, carazolol. For regulatory control purposes, tolerances for azaperone and carazolol have been set by the European Union as 100 and 25 [micro sign]g kg(-1), respectively. Furthermore, the use of the phenothiazines is prohibited and therefore has a zero tolerance. A method for the detection of residues of five tranquillisers and one [small beta]-blocker using a single ELISA plate has been developed. Kidney samples (2.5 g) were extracted with dichloromethane and applied to a competitive enzyme immunoassay using three polyclonal antibodies raised in rabbits against azaperol, propionylpromazine and carazolol conjugates. In sample matrix, the azaperol antibody cross-reacted 28.0% with azaperone and the propionylpromazine antibody cross-reacted 24.9% with acepromazine and 11.7% with chlorpromazine. In the ELISA, the detection capabilities of the six sedatives, azaperol, azaperone, carazolol, acepromazine, chlorpromazine, and propionylpromazine are 5, 15, 5, 5, 20 and 5 [micro sign]g kg(-1), respectively. The proposed method is a sensitive and rapid multi-residue technique that offers a cost effective alternative to current published procedures, without any concession on the ability to detect sedative misuse. PMID- 14752566 TI - Urinary tract infection and functional bladder sphincter disorders in children. AB - This review article focuses on the frequent association between urinary tract infection and functional bladder sphincter disorders which were distributed by the International Children's Continence Society in two main categories referred to as unstable bladder and dysfunctional voiding. Relevant examinations, including ultrasound, voiding cystourethrography, isotopic studies, and urodynamics, are described as well as their main advantages and drawbacks. Emphasis is put on the associated vesicoureteric reflux which should not be considered the central problem in those children. Reflux is induced by distortion of the ureteric meatus, a consequence of a long-standing high pressure in the bladder. Follow-up and treatment should aim to prevent renal damage and to protect the bladder function. PMID- 14752565 TI - Optimal mechanisms for finding and selecting mates: how threespine stickleback ( Gasterosteus aculeatus) should encode male throat colors. AB - Male threespine stickleback ( Gasterosteus aculeatus) use nuptial colors to attract mates and intimidate rivals. We quantified stickleback color and environmental lighting using methods independent of human perception to evaluate the information transmitted by male signals in a habitat where these signals are displayed. We also developed models of chromatic processing based on four cone photopigments (peak absorptions at 360, 445, 530, and 605 nm) characterized microspectrophotometrically in G. aculeatus and three other stickleback species. We show that a simple opponent mechanism receiving equally weighted inputs from cones with peak absorptions at 445 nm and 605 nm efficiently encodes variation in male throat colors. An orthogonal opponent mechanism-the difference between outputs of 530-nm cones and mean of outputs of 445- and 605-nm cones-produces a neural signal that could be used for species recognition and would be largely insensitive to variation in male throat color. We also show that threespine stickleback throats/photopigments are optimized for this coding scheme. These and other findings lead to testable hypotheses about the spectral processing mechanisms present in the threespine stickleback visual systems and the evolutionary interactions that have shaped this signal/receiver system. PMID- 14752567 TI - Ultrasonography of musculoskeletal infections in children. AB - Ultrasonography is an important modality for evaluation of musculoskeletal infections in children because it is rapid, nonionising and very sensitive for (infectious) fluid collections and joint effusions. Moreover, the images are not degraded by metallic or motion artefacts (as with CT and MRI) and finally, ultrasonography offers the possibility of fine-needle aspiration to confirm the infectious nature of a fluid collection without unnecessary contamination of adjacent anatomical compartments. Ultrasonography should be combined with radiography because both imaging techniques are complimentary. The purpose of this article is to emphasise the role of ultrasonography in the diagnosis of various diseases in childhood, including cellulitis, subcutaneous abscess, necrotizing fasciitis, pyomyositis, infectious bursitis and arthritis, osteomyelitis, foreign bodies and infectious lymphadenitis. Along with conventional radiography, ultrasonography is a very valuable modality for early diagnosis and follow-up of musculoskeletal infections in children. PMID- 14752568 TI - Urinary tract infection in infants and children: an update with special regard to the changing role of reflux. AB - The aim of this study was to present current theories of pathogenesis and prognosis in urinary tract infection (UTI) and renal scarring during infancy and childhood, with special regard to new insights concerning the role of vesico ureteral reflux (VUR). For a long time VUR and UTI were considered the only risk factors for renal scarring in childhood. Now a wider spectrum of contributing conditions is commonly accepted, which all may pose different clinical consequences and require different imaging approaches. Particularly bilateral renal scarring causes long-term sequalae; therefore, renal involvement in UTI with potential scarring has become the clinical and imaging focus, and proper diagnosis of UTI as the key factor for further management has become even more important. The VUR still remains one issue on a list of important factors such as treatment onset and response, bacterial virulence, immunological factors, genetic disposition, anatomical variants, and lower urinary tract dysfunction. Recent advances in knowledge leading to changed patho-physiological concepts, and new imaging techniques, may consecutively impact the presently established standard imaging algorithms. New, advanced imaging techniques offer improved and accelerated comprehensive imaging of the paediatric urinary tract. At present, this is complimentary to the established gold standard techniques. Strong research efforts have to be made before suggesting significant changes of current imaging concepts; however, based on recent technical advances and new insight on the natural history of paediatric urological diseases, potential changes of established imaging algorithms need to be discussed and evaluated. PMID- 14752569 TI - Current imaging concepts in pediatric osteomyelitis. AB - The diagnosis of osteomyelitis remains a difficult diagnostic dilemma. In this article, which is particularly aimed at those whose practice does not include a large paediatric population, we review the pathophysiology of paediatric osteomyelitis and contrast it with the available imaging modalities. We examine the role of the radiologist as well as the usefulness of each modality. Secondly, we review the different clinical scenarios such as acute, subacute and chronic, as well as specific forms of osteomyelitis; the latter includes subacute chronic ostemyelitis, toxic synovitis, spondylodiscitis as well as the congenital inflammatory disorders such as rubella and syphylis. The most useful imaging findings to look for and their significance are assessed and we evaluate their usefulness in each case. Close cooperation between clinicians and imagers remains the key to early and adequate diagnosis of paediatric osteomyelitis. PMID- 14752570 TI - Management of intussusception. AB - Intussusception represents the most common abdominal emergency in infancy. The classical clinical triad, consisting of abdominal colics, red jelly stools and a palpable mass, is only present in approximately 50% of cases, 20% of patients are symptom free at clinical presentation. Primary imaging modality of choice is ultrasound scanning, which enables the diagnosis or exclusion of an intussusception at a sensitivity of 98-100%, specificity of 88% and a negative predictive value of 100%. In emergency cases, additional plain films are necessary to detect potential intestinal perforation, to identify intestinal obstruction or other diseases mimicking the clinical presentation. Once the diagnosis of an intussusception is established, non-surgical reduction (NSR) is used. A surgical approach is chosen in patients with signs of perforation, shock or peritonitis. Depending on the choice of guiding imaging technique, different contrast media are used for NSR. Barium suspension or air with fluoroscopic guidance, or saline only or mixed with water-soluble contrast under sonographic guidance, has to be used. Regardless of the used contrast medium, NSR is an effective technique, being successfully employed in more than 90% of cases. PMID- 14752571 TI - HRCT in small and large airways diseases. AB - High-resolution computed tomography (HRCT) is being increasingly used in the diagnostic work-up of paediatric patients with large and small airways disease due to its ability to provide valuable information far beyond that of other non invasive investigations. This article highlights the key HRCT appearances of a range of conditions involving the airways in children, and where relevant, the role of HRCT in assessment of disease severity and monitoring of disease progression. PMID- 14752572 TI - Tumors and tumor-like lesions of the neck: from childhood to adult. AB - In childhood, tumors and tumor-like lesions of the neck are rare and tend to be benign. Very common are congenital cystic lesions (thyroglossal duct cysts, branchial cysts, dermoid cysts, lymphangiomas, cystic hygromas). Common neoplastic lesions are hemangiomas and papillomas. The most common malignant tumors in the head and neck region are lymphomas and rhabdomyosarcomas. Lymph node enlargements, reactive or/and infectious, account also for a significant amount of cervical masses. PMID- 14752573 TI - Ultrasound of female genital anomalies. AB - Congenital anomalies of the female genital tract result from mullerian duct anomalies and/or abnormalities of the urogenital sinus or cloaca. Due to the close developmental relationship between the genital and the urinary tracts, association of anomalies in both systems are common. This article reviews the appearance of developmental anomalies of the female urinary and genital tracts and points out common associated malformations to allow an early and complete sonographic assessment of affected infant. PMID- 14752574 TI - Radiology of diffuse interstitial pulmonary disease in children. AB - Diffuse interstitial lung disease (DILD) represents a heterogeneous group of disorders characterised by restrictive lung function and impaired gas exchange. As these diseases occur on a background of the developing lungs and immune system, the clinical presentation and disease progression is modified by comparison with their adult equivalents; thus, often differs markedly in presentation, clinical features and progress from ILD in adults, and it is not safe to extrapolate from adults to children. It is important to understand the normal growth and development of the lungs in children to understand the development of interstitial lung disease. As the chest radiograph is often non specific, HRCT has been shown in adults and children to increase the accuracy at diagnosis of diffuse lung disease. The trade-off in sensitivity and specificity of HRCT over chest X-ray is related to radiation dose which is significantly higher with conventional spiral or volumetric CT; however, the use of low-dose (50 mA, 0.75 s) limited (1-mm slices every 15-20 mm) HRCT in inspiration with three expiratory supplementary scans allows accurate assessment of the presence and extent of diffuse lung disease at a dose equivalent to approximately ten chest radiographs. Images are reconstructed on a high spatial resolution algorithm and displayed with a wide window setting, at a width of 1500 Hounsfield units (HU) and at a level of -500 HU. PMID- 14752575 TI - Urinary tract infection: an update on imaging strategies. AB - Urinary tract infections (UTIs) are a common cause of morbidity in children. The distinction between upper and lower tract UTI is clinically difficult but important, as permanent renal damage can occur when the UTI involves the kidneys. Children with a history of UTI need investigated promptly and thoroughly. The goal of imaging in UTI is to aid in the diagnosis of acute pyelonephritis, to identify those children who are at high risk of developing permanent renal damage, and to confirm and monitor the presence of renal scarring. This article reviews the role of radiology in each of these aims. PMID- 14752576 TI - Imaging strategies in children with suspected appendicitis. AB - Appendicitis is the commonest acute surgical emergency of childhood, and occurs in approximately 2-4 per 1000 infants. It is usually seen in infants older than 5 years but can occur at any age. Atypical clinical findings are seen in 30-50% of children, especially the younger ones, and often leads to a delayed diagnosis. Preoperative clinical assessment alone has yielded an accuracy ranging between 70 and 90% but is associated with a normal appendectomy rate of 13-25%. Preoperative imaging using the graded compression US technique and/or different helical CT techniques has been able to reduce this rate to 3-7% without an increase in perforation rate. An extensive review of the literature revealed several papers examining the accuracy of different imaging modalities and strategies of acute appendicitis in children. The reported sensitivity of US varied between 87 and 95%, vs 95-97% for helical CT, while the specificity ranged between 85 and 98% for US and 94 and 97% for helical CT. Only one truly randomised study was found, showing that compared with US alone, a combination of US and helical CT increased the sensitivity from 86 to 99%, while the specificity decreased from 95 to 89%. We conclude that imaging should be performed in all children with suspected appendicitis and that US should be the initial procedure with CT as a complementary tool. PMID- 14752577 TI - Abdominal pain: general imaging strategies. AB - Acute abdominal pain is the most common condition necessitating surgical admission to a paediatric hospital. The vast majority of cases are due to either appendicitis or acute non-specific abdominal pain; however, there are many other conditions presenting with acute abdominal pain in childhood which can mimic appendicitis. Some of these conditions are rarely encountered in adult practice. Plain abdominal radiographs are still obtained initially on many children presenting with abdominal pain. In the absence of associated symptoms such as vomiting however, ultrasonography (US) is more likely to elucidate the underlying cause of pain than plain-film radiography. While computed tomography (CT) is being used extensively in investigating adults with abdominal pain, its widespread use in children is to be avoided in the interest of radiation protection. PMID- 14752578 TI - Dexamethasone as a chemoprotectant in cancer chemotherapy: hematoprotective effects and altered pharmacokinetics and tissue distribution of carboplatin and gemcitabine. AB - PURPOSE: Hematoprotective strategies may offer new approaches to prevent chemotherapy-induced hematotoxicity. The present study was undertaken to investigate the chemoprotective effects of dexamethasone and its optimal dose and the underlying mechanisms. METHODS: Lethal toxicity and hematotoxicity of carboplatin were compared in CD-1 mice with or without dexamethasone pretreatment. Plasma and tissue pharmacokinetics of carboplatin were determined in CD-1 mice. Carboplatin was quantified by HPLC. Gemcitabine was analyzed by radioactivity counting. RESULTS: Pretreatment with dexamethasone prevented lethal toxicity of carboplatin in a dose- and schedule-dependent manner. The best protective effects of dexamethasone pretreatment as measured by survival were observed at the dose level of 0.1 mg/mouse per day for 5 days (80% vs 10% in controls). In contrast, posttreatment with dexamethasone had no protective effects. Pretreatment with dexamethasone significantly prevented the decrease in granulocyte counts. To elucidate the mechanisms by which dexamethasone pretreatment reduces hematotoxicity, we examined the effects of dexamethasone pretreatment on the pharmacokinetics of carboplatin and gemcitabine in CD-1 mice. No significant differences in plasma pharmacokinetics of carboplatin or gemcitabine were observed between control and mice pretreated with dexamethasone. However, dexamethasone pretreatment significantly decreased carboplatin and gemcitabine uptake in spleen and bone marrow with significant decreases in AUC, T(1/2), and C(max), and an increase in CL. CONCLUSIONS: To our knowledge, this is the first time that dexamethasone has been shown to significantly decrease host tissue uptake of chemotherapeutic agents, suggesting a mechanism responsible for the chemoprotective effects of dexamethasone. This study provides a basis for future study to evaluate dexamethasone as a chemoprotectant in cancer patients. PMID- 14752579 TI - Metabolic selectivity and growth of Clostridium thermocellum in continuous culture under elevated hydrostatic pressure. AB - The continuous culture of Clostridium thermocellum, a thermophilic bacterium capable of producing ethanol from cellulosic material, is demonstrated at elevated hydrostatic pressure (7.0 MPa, 17.3 MPa) and compared with cultures at atmospheric pressure. A commercial limitation of ethanol production by C. thermocellum is low ethanol yield due to the formation of organic acids (acetate, lactate). At elevated hydrostatic pressure, ethanol:acetate (E/A) ratios increased >10(2) relative to atmospheric pressure. Cell growth was inhibited by approximately 40% and 60% for incubations at 7.0 MPa and 17.3 MPa, respectively, relative to continuous culture at atmospheric pressure. A decrease in the theoretical maximum growth yield and an increase in the maintenance coefficient indicated that more cellobiose and ATP are channeled towards maintaining cellular function in pressurized cultures. Shifts in product selectivity toward ethanol are consistent with previous observations of hydrostatic pressure effects in batch cultures. The results are partially attributed to the increasing concentration of dissolved product gases (H2, CO2) with increasing pressure; and they highlight the utility of continuous culture experiments for the quantification of the complex role of dissolved gas and pressure effects on metabolic activity. PMID- 14752580 TI - Molecular cloning and characterization of the polymorphic MHC class II DBB from the tammar wallaby (Macropus eugenii). AB - Genes of the major histocompatibility complex (MHC) have been characterized in all extant lineages of mammals. The tammar wallaby (Macropus eugenii) is well established as a model marsupial species; however, no classical MHC sequences have been described from this species. We have isolated two MHC class II beta chain sequences from a tammar wallaby spleen cDNA library using a tammar MHC class II beta probe. These sequences belong to the marsupial MHC class II DBB gene family. Two additional DBB sequences were amplified from tammar wallaby genomic DNA. All four sequences were obtained from the same individual, indicating that there are at least two DBB loci in the tammar wallaby. PMID- 14752581 TI - Nimodipine does not affect the flow-metabolism couple in permanent cerebral ischemia. AB - Despite widespread investigational and clinical usage of the calcium channel blocker nimodipine, its effects on cerebral physiology in normal and ischemic brain remain poorly understood. In order to gain insight into this subject we examined the effects of nimodipine on glucose metabolism and cerebral blood flow metabolism coupling in the rat during conditions of reproducible focal ischemia. Nimodipine-treated animals were then matched with vehicle-treated controls for both study conditions. Animals were subjected to permanent occlusion of the middle cerebral artery (MCA) along with occlusion of the common carotid arteries. Five minutes into ischemia, an intravenous infusion of nimodipine (1 micro g/kg per min, n=9) or vehicle ( n=9) was initiated and continued until the end of the study. Seventy-five minutes after the occlusion, [14C]2-deoxyglucose was injected into the venous catheter for the measurement of the local cerebral metabolic rate for glucose (LCMRgl), followed 25 min later by the injection of N-isopropyl [123I] p-iodoamphetamine for the measurement of local cerebral blood flow (LCBF). The animals were killed at the end of 2 h of ischemia, and the brains were processed for double-labeled autoradiography. In all animals, permanent MCA occlusion produced significant decrements in LCBF, LCMRgl, and LCBF/LCMRgl ratio in both the core of the ischemia as well as regions peripheral to the ischemia within the same cerebral hemisphere when compared with non-ischemic brain. There were no significant differences between the nimodipine-treated and vehicle treated groups. In conclusion, nimodipine does not appear to alter cerebral blood flow or cerebral metabolism in ischemic brain. PMID- 14752583 TI - Determination of organotin compounds in biological samples using accelerated solvent extraction, sodium tetraethylborate ethylation, and multicapillary gas chromatography-flame photometric detection. AB - A method has been developed for species-selective analysis of organotin compounds in solid, biological samples. The procedure is based on accelerated solvent extraction (ASE) of analytes and includes extraction of the tin species with a methanol-water (90% methanol) solution of acetic acid/sodium acetate containing tropolone (0.03% w/ v), their ethylation with NaBEt(4), and separation and detection by GC-FPD. The analytical procedure was optimized with an unspiked sample of harbor porpoise ( Phocoena phocoena) liver. Effects of ASE operational variables (extraction temperature and pressure, solvent composition, number of static extraction steps) are discussed. Method detection limits (MDL) were in the range 6-10 ng(Sn) g(-1) dry weight and 7-17 ng(Sn) g(-1) dry weight for butyl- and phenyltin compounds, respectively. Recoveries were comparable with or better than those obtained by use of other procedures reported in the literature. The analytical procedure was validated by analysis of NIES No. 11 (fish tissue) certified reference material. PMID- 14752585 TI - NMR study of some diastereomeric nucleoside-3'-thiophosphate derivatives in solution. AB - The conformational properties of diastereomeric P-modified nucleotides are reported as reflected by different NMR parameters. Some conformational trends can be rationalized by consideration of the (3) J(C4',P)()and (3) J(C2',P) coupling values of the isomers and the nature of the substituent on the phosphorus. Configurational assessment of the phosphorus is inferred from NOE experiments. The effects of temperature, solvent and size of substituents are presented. PMID- 14752582 TI - Spatio-temporal luminance contrast sensitivity and visual backward masking in schizophrenia. AB - The aim of two experiments was to investigate the relationship between spatio temporal contrast sensitivity and visual backward masking in normal observers and in subgroups with positive or negative symptoms in schizophrenia. Experiment 1 measured contrast sensitivity for stationary and counterphase-modulated sinusoidal gratings at four spatial (0.5, 2.0, 4.0, 8.0 cycles/degree) and four temporal frequencies (0, 4.0, 8.0, 16.0 Hz). The results showed that there were no differences in spatio-temporal contrast sensitivity between the control and positive-symptom group, and in comparison with these groups, contrast sensitivity was significantly lower at all spatial and temporal frequencies in the negative symptom group. Experiment 2 measured the visibility of a Landolt C target with a constant target stimulus duration of 4.0 ms followed by a 150-ms backward mask, which was presented at 12 stimulus onset asynchronies from 0 to 110 ms in the same groups of observers. Consistent with the findings of the previous experiment, there were no significant differences in backward masking between the control and positive-symptom group, and in comparison with these groups, visual backward masking was significantly higher at all stimulus onset asynchronies from 40 to 110 ms in the negative-symptom group. The present findings show that there were no significant differences in contrast sensitivity and in backward masking between normal observers and a group with positive symptoms in schizophrenia. It was concluded that the reduction in contrast sensitivity for low spatial frequency counterphase flicker in the negative-symptom group is consistent with a reduction in the 'contrast gain control' mechanism of magnocellular channels, and that the reduction in contrast sensitivity for medium and high stationary gratings is consistent with a disorder in parvocellular channels. It was proposed that a disorder in magnocellular channels in the negative-symptom group may enforce a reliance on parvocellular channels that results in longer temporal summation and visible persistence, slower visual processing of single target stimuli at threshold and higher levels of sensory integration, and backward masking when multiple stimuli are presented rapidly in time. PMID- 14752586 TI - Effects of morphine and naltrexone on impulsive decision making in rats. AB - RATIONALE: It has been reported that human opiate addicts discount delayed rewards more than non-addicts, indicating that they are more impulsive. However, it is not clear whether this difference reflects pre-existing traits, or the effects of exposure to the opiates. OBJECTIVES: This study was designed to investigate the effects of an opioid agonist and antagonist on delay discounting in rats. The study had three objectives: to determine (1) the acute effects of the opioid agonist morphine (MOR) on delay discounting, (2) the acute effects of the opioid antagonist naltrexone (NAL) on delay discounting, and (3) whether NAL reverses the effects of MOR on delay discounting. METHODS: An adjusting amount procedure (AdjAmt) was used to determine how much animals discounted the value of delayed rewards. Acute doses of MOR (0.3, 1.0, and 1.8 mg/kg SC), NAL (0.01, 0.1, 1.0, and 10 mg/kg SC) and NAL (0.1 mg/kg SC) prior to MOR (1.8 mg/kg SC) were tested in 15 rats. RESULTS: MOR dose dependently increased the rate of delay discounting (i.e., made the animals more impulsive). NAL alone had no effect on the value of delayed rewards, but NAL blocked the effects of MOR. CONCLUSIONS: These results suggested that the direct effects of MOR may contribute to the high level of impulsive behavior seen among opiate users. PMID- 14752588 TI - Decreased pain tolerance and mood in recreational users of MDMA. AB - RATIONALE: 3,4-Methylenedioxymethamphetamine (MDMA) is known to affect brain serotonin (5-HT) neurons in experimental animals. However, its effects on humans are more difficult to infer. Serotonin is implicated in the body's ability to modulate the effects of pain and to regulate mood. OBJECTIVE: The aim of this research is to test nociceptive responses and mood in MDMA users as an index of central 5-HT function. METHOD: Measurements of pain tolerance were obtained using the cold-pressor test for 15 polydrug users who regularly use MDMA, 3-4 days after the most recent usage, and ten matched polydrug users who do not use MDMA. A rating on mood was obtained for each participant using the Nowlis Mood Adjective checklist. RESULTS: Measurements of pain tolerance and mood were significantly lower in the MDMA group. A positive correlation between mood and pain tolerance was found in the MDMA group, whereas no correlation was found between these variables in the non-MDMA group CONCLUSION: This study found an association between pain tolerance and MDMA usage and confirmed the association between MDMA and depressed mood. The current results suggest that MDMA, at least in the short term, may cause serotonin-mediated alterations in pain sensitivity. PMID- 14752589 TI - Screening for highly active beta-lactam antibiotics against Agrobacterium tumefaciens. AB - Quantitative in vitro antibacterial activities, i.e., minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBCs), of 12 beta lactam antibiotics against Agrobacterium tumefaciens strains LBA4404 and EHA101 were examined, in order to identify antibiotics effective in eliminating the bacteria in Agrobacterium-mediated plant genetic transformation. The antibacterial activities of beta-lactams tested against strain EHA101 were equal to or less than those tested against strain LBA4404. Cefotaxime, cefbuperazone, and meropenem had high activities against strain LBA4404 (MBC <1 mg l(-1)). Against strain EHA101, however, only meropenem showed activity comparable to that against strain LBA4404. The production of beta-lactamase was observed only in strain EHA101. PMID- 14752590 TI - What is the clinical relevance of a paravaginal defect? PMID- 14752591 TI - Diagnosis of cystocele--the correlation between clinical and radiological evaluation. AB - In patients with genital prolapse involving several compartments simultaneously, radiologic investigation can be used to complement the clinical assessment. Contrast medium in the urinary bladder enables visualization of the bladder base at cystodefecoperitoneography (CDP). The aim of the present study was to evaluate the correlation between clinical examination using the Pelvic Organ Prolapse Quantification system (POP-Q) and CDP. Thirty-three women underwent clinical assessment and CDP. Statistical analysis using Pearson's correlation coefficient ( r) demonstrated a wide variability between the current definition of cystocele at CDP and POP-Q ( r=0.67). An attempt to provide an alternative definition of cystocele at CDP had a similar outcome ( r=0.63). The present study demonstrates a moderate correlation between clinical and radiologic findings in patients with anterior vaginal wall prolapse. It does not support the use of bladder contrast at radiologic investigation in the routine preoperative assessment of patients with genital prolapse. PMID- 14752592 TI - Prevalence of urinary, fecal and double incontinence in the elderly living at home. AB - The aim of this study was to evaluate the prevalence of urinary, fecal and double incontinence in the elderly, through a population-based cross-sectional survey. The study included all patients aged 60 and over of nine general practices in the Nijmegen Monitoring Project. Patients living in a home for the elderly were excluded, as well as patients with dementia, patients who were too ill to participate and patients with a catheter. There were 5278 selected patients who received a postal questionnaire. Of these, 88% returned it. Nineteen percent of the respondents had involuntary loss of urine twice a month or more, 6% loss of feces and 3% both. The prevalence of urinary, fecal and double incontinence increased with age in both men and women, and especially in men in the oldest age group. Urinary incontinence was more prevalent in women than in men. The prevalence of fecal incontinence showed no sex differences, but the type of fecal incontinence did differ between men and women. In men loss of mucus was twice as common as in women. Double incontinence was also equally prevalent in men and women, except in the age group 65-74 years. In conclusion, urinary, fecal and double incontinence are common conditions in the community-dwelling population. The prevalence rates increase with age. Urinary incontinence is more prevalent in women. There were no sex differences in the prevalence of fecal incontinence but the type of fecal incontinence was different in men and women. PMID- 14752594 TI - Mini-incisional Burch urethropexy: a less invasive method to accomplish a time tested procedure for treatment of genuine stress incontinence. AB - The purpose of this study was to evaluate the feasibility, safety and efficacy of performing the Burch urethropexy (BU) and the abdominal paravaginal repair (APR) through a 1.5-2.5 in suprapubic incision. A prospective clinical study was undertaken by four urogynecologists. Seventy-three patients, each with a urodynamic and clinical diagnosis of genuine stress incontinence, underwent a BU procedure, with 33 of the 73 having concomitant APR through the same incision. The duration of surgery and any complications were recorded. Postoperative outcome tests included subjective incontinence questionnaire, cough stress testing, pad testing, measurement of residual volumes, and analgesia requirements. The BU procedure was accomplished in 72 of 73 patients, with 1 requiring conversion to a 5 in incision. The mean operative time was 64.6 +/-21.9 (SD) min. Intraoperatively, 1 patient was noted to have a suture in the bladder. All patient having only a BU (40) went home on the day of surgery or the first postoperative day, and all patients with BU and APR went home within 2 days. All but 1 patient met the criteria for catheter removal within 7 days, with 1 patient suffering obstructive voiding. At a mean follow-up of 9 months, cough stress test and questionnaire demonstrated complete cure in 70 of 72 patients tested. Pad testing confirmed cure in all of the 46 patients who consented to the test. We conclude that the standard Burch procedure and paravaginal repair can be accomplished safely and with excellent short-term efficacy through a 1.5-2.5 in incision. PMID- 14752593 TI - The short-term prevalence of de novo urinary symptoms after different modes of hysterectomy. AB - The aim of this study was to determine the short-term prevalence of de novo urinary symptoms after hysterectomy indicated by meno/metrorrhagia or dysmenorrhea/dyspareunia. The study group consisted of 451 women who had had a hysterectomy for reasons of meno/metrorrhagia or dysmenorrhea/dyspareunia. Fifty three (12%) had a supracervical, 151 (33%) a total abdominal and 247 (55%) a vaginal hysterectomy. As a non-gynecologic background population we enrolled 110 women who had had their gallbladder removed laparoscopically. All women received a postal questionnaire 9-45 months after their operation. Specific questions were asked about their voiding habits, comprising significant stress incontinence, bothersome stress incontinence, significant urge incontinence, bothersome urge incontinence, pollakisuria, nocturia, use of pads, and the feeling of having a hygiene problem. To evaluate de novo symptoms or de novo cure, the women assessed the symptoms before as well as after the operation. Results showed that abdominal hysterectomy lasted longer, had heavier blood loss and required longer hospitalization than did vaginal or supracervical hysterectomy. Women scheduled for a supracervical hysterectomy had preoperatively more significant and bothersome urge incontinence, and postoperatively more significant urge, urgency, and feeling of having a hygienic problem than did women having a vaginal hysterectomy, a total abdominal hysterectomy or a laparoscopic cholecystectomy. When assessing de novo symptoms, supracervical hysterectomy was associated with more urgency and the feeling of having a hygienic problem. Some women experienced de novo cure, but these were almost exclusively in the study group and rarely in the control group. It was concluded that supracervical hysterectomy is related to more urinary symptoms than vaginal or total abdominal hysterectomy. De novo symptoms as well as de novo cure are common, which is why urinary symptoms after hysterectomy must be evaluated over time. PMID- 14752595 TI - Voiding dysfunction after surgery for stress incontinence: literature review and survey results. AB - Postoperative voiding dysfunction is a potential complication of anti incontinence procedures. Reported rates of urethral obstruction range from 5% to 20%. There is a lack of consensus in the literature regarding the appropriate evaluation and management of this distressing problem. A literature search was carried out using Medline (1966-2001) for postoperative voiding dysfunction. The key word urethrolysis was cross-referenced with surgical complications and stress urinary incontinence to identify all published English-language articles. The bibliographies of reviewed articles were searched manually. We also mailed a survey to the members of American Urogynecologic Society (AUGS) regarding their management of this problem. Overall, 262 members (31.4%) responded to the survey. Success rates reported in the literature between retropubic and vaginal techniques of urethrolysis are comparable, but morbidity is lower with the vaginal approach. The success rates are equivalent with (68%) or without (74%) resuspension following transvaginal urethrolysis. The incidence of postoperative SUI is acceptably low even without resuspension of the urethra (6% for both). Results of the AUGS survey reveal that most providers favor a transvaginal approach (74%) when performing urethrolysis, and they do not routinely resupport the bladder neck (82%). PMID- 14752596 TI - The effects of the tension-free vaginal tape on voiding function: a prospective evaluation. AB - The impact of tension-free vaginal tape (TVT) on voiding function was studied in a cohort of 103 patients. Voiding was assessed with quality of life questionnaires, non-invasive urine flow and voiding pressure-flow studies both preoperatively and 1 year postoperatively. Paired t-tests and Wilcoxon's signed ranks tests were used for analysis. Data from 65 patients were analyzed. Subjective voiding function did not change; however, maximum flow rates decreased by up to 43%: from 29 ml/s to 16 ml/s on free flow studies. This was associated with a 27% increase in detrusor pressure at maximum flow: from 15 to 19 cmH(2)O. Postvoid residuals were not clinically different, changing from a median of 15 ml to 30 ml postoperatively. Thirty-eight patients (37%) required postoperative catheterization for urinary retention (median duration 4 days). We concluded that the TVT sling reduces maximum flow rates without clinically significant effects on detrusor pressure, residual urine or subjective voiding. PMID- 14752597 TI - Initial experience with a new method for the dynamic assessment of pelvic floor function in women: the Kolpexin Pull Test. AB - The aim of this study was to evaluate a newly developed, simple test for assessing pelvic floor muscular function in women. Pelvic floor strength in women with symptomatic pelvic floor dysfunction was evaluated with a newly developed assessment tool, the Kolpexin Pull Test, and compared to a clinically validated digital pelvic floor strength assessment scale (DPA). A 36-mm Kolpexin sphere was inserted into the vagina above the levator plate and connected to a digital tensiometer/force gauge. The force required to remove the sphere was recorded for three resting trials and three maximum pelvic floor contractions. Results of the DPA and Kolpexin Pull Test were analyzed using ANOVAs of contrast variables, intraclass correlations, and regression analyses. Twenty-one women participated in the study. Age range was 36-85 years, parity range was 2-5. All six trials required less than 5 min to perform in each patient. Intraperson maximum contraction data were correlated at 0.96 (95% CI: 0.91, 0.98), and were greater at higher DPA scores ( p=0.016). There were positive correlations between the maximum contraction and DPA data (adjusted R(2)=0.52; p <0.001), and the maximum contraction minus resting vs DPA data (adjusted R(2)=0.54; p< 0.001). The Kolpexin Pull Test is reproducible, rapid, and correlates with digital clinical assessment of pelvic floor strength during maximal contractions. PMID- 14752598 TI - The position of the urethrovesical junction after incontinence surgery: early postoperative changes. AB - This study prospectively evaluated the position of the urethrovesical junction using the Q-tip angle to assess early postoperative changes for different anti incontinence surgeries. All procedures resulted in a statistically significant change in resting angle from the intraoperative value. The mean change for the transvaginal tape was 25.74 degrees (27.43 to 3.28); Burch 11.18 degrees (-20.44 to -10.0) and fascia sling 13.9 degrees (26.57 to 15.68). The mean change in Q tip angle was greater after transvaginal tape placement than after Burch ( p=0.000) and fascial sling ( p=0.022) procedures. These findings show that the resting position of the urethrovesical junction after surgery is different for all procedures. The transvaginal tape results in the greatest change in angle. This may help to negate the so-called 'tension-free' nature of the procedure. Surgeons need to be aware of this, as it may be an etiological factor in cases of late urinary retention and urethral erosion. PMID- 14752599 TI - Acute renal failure caused by complete bladder eversion through a vesicovaginal fistula. AB - We report a rare case of bladder eversion through a vesicovaginal fistula. The bladder prolapse was almost complete, resulting in ureteral kinking, bilateral hydronephrosis and acute renal failure. After reduction of the bladder eversion, bilateral ureteral stent placement, fistula repair using the Latzko technique and colpocleisis, the patient had rapid resolution of her renal compromise. PMID- 14752600 TI - Sliding hernia of the bladder presenting as suprapubic pain in a female. AB - We report a case of bilateral sliding hernia of the bladder in a 64-year-old woman who presented with suprapubic pain and nocturia and was diagnosed by voiding diary and cystogram. PMID- 14752601 TI - Delayed presentation and successful repair of a recurrent vesicovaginal fistula after hysterectomy and primary abdominal repair. AB - Urogenital fistulae are an uncommon consequence of gynecologic surgery. Vesicovaginal fistulae due to gynecologic surgery generally appear 1-6 weeks after surgery and recurrent fistulae within 3 months of their repair. The pathogenesis of vesicovaginal fistula formation remains unclear. We present the case of a 36-year-old woman with a spontaneously recurring vesicovaginal fistula 21 months after abdominal repair of a vesicovaginal fistula caused by a laparoscopic-assisted vaginal hysterectomy. During the repair of the fistula and excision of the vaginal cuff, two small fluid-filled cysts between the bladder mucosa and the vaginal epithelium were encountered. Vesicovaginal fistulae can occur spontaneously and remote from surgery. The finding of small fluid-filled cysts in the cuff suggests that rupture of a similar cyst may have led to the formation of the fistula. PMID- 14752602 TI - Suburethral vaginal erosion and pyogenic granuloma formation: an unusual complication of intravaginal slingplasty (IVS). AB - We report an unusual case of suburethral vaginal erosion and pyogenic granuloma formation 14 months after intravaginal slingplasty (IVS). A 64-year-old woman underwent IVS for recurrent stress incontinence 12 years after Burch colposuspension. Following seemingly uncomplicated surgery and recovery, she developed a recurrent urinary tract infection which was treated with antibiotics. When she presented with vaginal pain and postmenopausal bleeding approximately 14 months postoperatively, she was found to have suburethral vaginal erosion of the tape and a pyogenic granuloma. The exposed tape was removed, the granuloma excised, and the overlying vaginal skin was then closed. She then made an uneventful recovery. PMID- 14752603 TI - Gauze packing for aspirin-induced hemorrhage in vaginal hysterectomy. AB - Intra-abdominal packing has been used to control massive hemorrhage in many difficult situations. Gynecologists are finding it increasingly useful in controlling persistent hemorrhage in a variety of situations. Recently we found it necessary to use packing for persistent brisk bleeding during 'vaginal hysterectomy' in a patient on aspirin therapy for heart disease. We could find no similar report in the literature and now describe our experience. PMID- 14752604 TI - Effect of testosterone on lipolysis in human pre-adipocytes from different fat depots. AB - AIM/HYPOTHESIS: Regional differences in lipolysis, with higher lipolytic activity in visceral than subcutaneous fat, are important for the development of insulin resistance and might be influenced by testosterone. METHODS: We studied testosterone-regulated lipolysis and protein expression (by western blot) in fully differentiated pre-adipocytes from visceral (omental) and abdominal subcutaneous adipose tissue from 52 human subjects. These cells were isolated and cultured in a serum-free medium. RESULTS: Testosterone caused a specific, time- and concentration-dependent 50% reduction of catecholamine-stimulated lipolysis in the subcutaneous depot. Half of the maximum effect occurred at 10 nmol/l. The inhibitory effect was due to the inability of beta-adrenoceptors and cyclic AMP to stimulate the protein kinase A, hormone-sensitive lipase complex. Testosterone caused a depot-specific 50% reduction of the protein expression of hormone sensitive lipase and beta(2)-adrenoceptors in differentiated subcutaneous pre adipocytes, but no change in beta(1)-adrenoceptors, protein kinase A subunits or perilipin expression. In contrast, testosterone had no effect on lipolysis or protein expression in the visceral depot. However, testosterone receptors were present in both depots, and the hormone inhibited adipocyte leptin secretion. Similar effects on lipolysis were observed with dihydrotestosterone. CONCLUSIONS/INTERPRETATION: Testosterone in physiological concentrations causes a depot-specific reduction of catecholamine-stimulated lipolysis in subcutaneous fat cells, probably due to reduced protein expression of beta(2)-adrenoceptors and hormone-sensitive lipase. This could be an important pathogenic factor underlying regional differences in lipolysis and development of insulin resistance and hyperandrogenic polycystic ovary syndrome. PMID- 14752605 TI - Phylogenetic analysis of the highland papayas ( Vasconcellea) and allied genera (Caricaceae) using PCR-RFLP. AB - The chloroplast and mitochondrial DNA diversity of 61 genotypes belonging to 18 Vasconcellea species, the so-called highland papayas, was studied by PCR-RFLP analysis of two non-coding cpDNA regions ( trnM- rbcL and trnK1- trnK2) and one non-coding mtDNA region ( nad4/1- nad4/2). This sample set was supplemented with six genotypes belonging to three other Caricaceae genera: the monotypic genus Carica, including only the cultivated papaya, and the genera Jacaratia and Cylicomorpha. Moringa ovalifolia was added as an outgroup species. The PCR amplified cpDNA regions were digested with 18 restriction endonucleases, the mtDNA region with 11. A total of 22 point mutations and four insertion/deletions were scored in the sample. A higher level of interspecific variation was detected in the two cpDNA regions in comparison to the analysis of the mtDNA. Wagner parsimony and Neighbor-Joining analysis resulted in dendrograms with similar topologies. PCR-RFLP analysis supported the monophyly of Caricaceae, but among the 26 mutations scored, an insufficient number of markers discriminated between the different Caricaceae genera included in this study. Hence the inference of the intergeneric relationships within Caricaceae was impossible. However, some conclusions can be noted at a lower taxonomic level. The Caricaceae species were divided into two lineages. One group included only Vasconcellea spp., whereas the second included the remaining Vasconcellea spp., together with the papaya genotypes and those from the other Caricaceae genera. This may indicate a higher level of inter-fertility for the Vasconcellea species from the latter clade in interspecific crossings with papaya. The putative progenitors of the natural sterile hybrid V. x heilbornii, i.e. V. stipulata and V. cundinamarcensis, were only distantly related to V. x heilbornii. This indicates that probably none of these species was involved as the maternal progenitor in the origin of V. x heilbornii. Surprisingly, V. x heilbornii had organellar genome patterns identical with V. weberbaueri, suggesting a possible involvement of this species in the origin of V. x heilbornii. On the basis of discrepancy between morphological traits and the cpDNA profiles of some pairs of Vasconcellea species, we believe that besides V. x heilbornii, some other species have originated through interspecific hybridization. A reticulate evolution for Vasconcellea has therefore been suggested. Finally, intraspecific cpDNA variation was detected in V. microcarpa, thus providing molecular evidence for the high diversity previously indicated by morphological observations. PMID- 14752607 TI - Multivariate statistical model for 3D image segmentation with application to medical images. AB - In this article we describe a statistical model that was developed to segment brain magnetic resonance images. The statistical segmentation algorithm was applied after a pre-processing stage involving the use of a 3D anisotropic filter along with histogram equalization techniques. The segmentation algorithm makes use of prior knowledge and a probability-based multivariate model designed to semi-automate the process of segmentation. The algorithm was applied to images obtained from the Center for Morphometric Analysis at Massachusetts General Hospital as part of the Internet Brain Segmentation Repository (IBSR). The developed algorithm showed improved accuracy over the k-means, adaptive Maximum Apriori Probability (MAP), biased MAP, and other algorithms. Experimental results showing the segmentation and the results of comparisons with other algorithms are provided. Results are based on an overlap criterion against expertly segmented images from the IBSR. The algorithm produced average results of approximately 80% overlap with the expertly segmented images (compared with 85% for manual segmentation and 55% for other algorithms). PMID- 14752606 TI - The role of epithelial-to-mesenchymal transition in renal fibrosis. AB - Epithelial-to-mesenchymal transition (EMT) involving injured epithelial cells plays an important role in the progression of fibrosis in the kidney. Tubular epithelial cells can acquire a mesenchymal phenotype, and enhanced migratory capacity enabling them to transit from the renal tubular microenvironment into the interstitial space and escape potential apoptotic cell death. EMT is a major contributor to the pathogenesis of renal fibrosis, as it leads to a substantial increase in the number of myofibroblasts, leading to tubular atrophy. However, recent findings suggest that EMT involving tubular epithelial cell is a reversible process, potentially determined by the surviving cells to facilitate the repopulation of injured tubules with new functional epithelia. Major regulators of renal epithelial cell plasticity in the kidney are two multifunctional growth factors, bone morphogenic protein-7 (BMP-7) and transforming growth factor beta1 (TGF-beta1). While TGF-beta1 is a well established inducer of EMT involving renal tubular epithelial cells, BMP-7 reverses EMT by directly counteracting TGF-beta-induced Smad-dependent cell signaling in renal tubular epithelial cells. Such antagonism results in the repair of injured kidneys, suggesting that modulation of epithelial cell plasticity has therapeutic advantages. PMID- 14752609 TI - Brainprot - generating the proteome of the brain. PMID- 14752608 TI - Microcolinearity between a 2-cM region encompassing the grain protein content locus Gpc-6B1 on wheat chromosome 6B and a 350-kb region on rice chromosome 2. AB - The conservation of the linear order (colinearity) of genetic markers along large chromosome segments in wheat and rice is well established, but less is known about the microcolinearity between both genomes at subcentimorgan distances. In this study we focused on the microcolinearity between a 2.6-cM interval flanked by markers Xcdo365 and Xucw65 on wheat chromosome 6B and rice chromosome 2. A previous study has shown that this wheat segment includes the Gpc-6B1 locus, which is responsible for large differences in grain protein content (GPC) and is the target of a positional cloning effort in our laboratories. Twenty-one recombination events between Xcdo365 and Xucw65 were found in a large segregating population (935 gametes) and used to map 17 genes selected from rice chromosome 2 in the wheat genetic map. We found a high level of colinearity between a 2.1-cM region flanked by loci Xucw75 and Xucw67 on wheat chromosome 6B and a 350-kb uninterrupted sequenced region in rice chromosome arm 2S. Colinearity between these two genomes was extended to the region proximal to Xucw67 (eight colinear RFLP markers), but was interrupted distal to Xucw75 (six non-colinear RFLP markers). Analysis of different comparative studies between rice and wheat suggests that microcolinearity is more frequently disrupted in the distal region of the wheat chromosomes. Fortunately, the region encompassing the Gpc-6B1 locus showed an excellent conservation between the two genomes, facilitating the saturation of the target region of the wheat genetic map with molecular markers. These markers were used to map the Gpc-6B1 locus into a 0.3-cM interval flanked by PCR markers Xucw79 and Xucw71, and to identify five candidate genes within the colinear 64-kb region in rice. PMID- 14752610 TI - Metabolism and function in animal tissues of agmatine, a biogenic amine formed from arginine. AB - Recently agmatine, decarboxylated arginine, has been shown to be an important biological compound in several animal tissues. This paper summarizes the known information regarding the transport of arginine, its decarboxylation and the effects of the agmatine formed mainly on NO and polyamine synthesis. PMID- 14752611 TI - Evidence for the existence of hypothetical proteins in human bronchial epithelial, fibroblast, amnion, lymphocyte, mesothelial and kidney cell lines. AB - The human genome maybe limited to about 30000 genes whereas the human proteome may be represented by a rough estimate of one million proteins. A legion of proteins have been described and information about these structures are readily available in data banks. There remains, however, a large series of unknown or hypothetical proteins (HPs). Many of them have been predicted from nucleic acid sequences only and are therefore named predicted or HPs. Carrying out "protein hunting" by generating large maps of human cell lines, we aimed to find and identify HPs and provide an analytical tool thereof. Cell lysates from human bronchial epithelial, fibroblast, amnion, lymphocyte, mesothelial and kidney cell lines were prepared and proteins run on two-dimensional gel-electrophoresis (2DE) with in-gel digestion and mass spectrometrical analysis using the MALDI-TOF principle.16 HPs were found in these cell lines and some show cell-specific expressional patterns. HPs belong to several protein classes including structural, signaling, transcriptional/translational, chaperone-related and others. We furthermore provide analytical data i.e. pIs that were often different from predicted values in data banks.A list of HPs has been shown to really exist in several human cell lines thus contributing to knowledge on protein machineries and cascades. Observed and predicted pI values are given representing an analytical tool along with unambiguous identification of protein spots by mass spectrometry independent of antibody availability and specificity thus complementing established methods. PMID- 14752612 TI - Inhibition of the L-dopa transport system in human epidermal Langerhans cells by omeprazole and its analogues. AB - L-3,4-dihydroxyphenylalanine (L-dopa) transport into human Langerhans cells (LC) occurs by a saturable mediation. This plasma membrane agency is, due to its characteristics, distinguishable from systems transporting other neutral, cationic and anionic amino acids into other cells and serves to catalyze the flow of L-dopa, only, into LC. The uphill operation of this L-dopa transport system is believed to occur by down-gradient countermigration of H(+). Due to the uniqueness of the L-dopa transport system, the widely used analogue inhibition approach was not applicable. Instead we studied omeprazole and its analogues in our search for suitable inhibitory candidates. Omeprazole and most of its analogues were indeed inhibitory in the concentration range 1-100 micromol/L. Conspicuously, the compounds with strongest polarity were least inhibitory. The inhibitory pattern displayed by omeprazole and the other analogues on L-dopa uptake in LC corresponded to some extent to what has been observed previously for purified H(+),K(+)-ATPase from tubulovesicles of the stomach. No effects of the inhibitors were registered on energy charge and lactate production of epidermal biopsies, nor were any gross alterations of ultrastructure of LC noticed. PMID- 14752613 TI - Fractionation of liver proteins by preparative electrophoresis. AB - Proteomics offers unique possibilities to investigate changes in the levels and modifications of proteins involved in the pathomechanisms of diseases and toxic events. However, search for potential drug targets and disease or toxicity markers is limited by the fact that mainly the high-abundance, hydrophilic proteins are visualized in two-dimensional gels. Here we studied the enrichment of rat liver cytosolic proteins by preparative electrophoresis. Preparative electrophoresis was performed with the PrepCell apparatus in the presence of 0.1% lithium dodecyl sulfate. Lithium dodecyl sulfate was exchanged against agents compatible with isoelectric focusing prior to the two-dimensional gel electrophoresis. Proteins were identified from two-dimensional gels by matrix assisted laser desorption ionization time-of-flight mass specrometry. Low- and middle-size proteins and low-abundance proteins, which had not been found before, were enriched by preparative electrophoresis. The present study represents a contribution of proteomics in the quantification of differences in the levels of low-abundance liver proteins in toxicity studies. PMID- 14752614 TI - Nitric oxide is not involved in the control of vasopressin release during acute forced swimming in rats. AB - Neurons of the hypothalamo-neurohypophyseal system (HNS) are known to contain high amounts of neuronal nitric oxide (NO) synthase (nNOS). NO produced by those neurons is commonly supposed to be involved as modulator in the release of the two nonapeptides vasopressin (AVP) and oxytocin into the blood stream. Previous studies showed that forced swimming fails to increase the release of AVP into the blood stream while its secretion into the hypothalamus is triggered. We investigated here whether hypothalamically acting NO contributes to the control of the AVP release into blood under forced swimming conditions. Intracerebral microdialysis and in situ hybridization were employed to analyze the activity of the nitrergic system within the supraoptic nucleus (SON), the hypothalamic origin of the HNS. A 10-min forced swimming session failed to significantly alter the local NO release as indicated both by nitrite and, the main by-product of NO synthesis, citrulline levels in microdialysis samples collected from the SON. Microdialysis administration of NO directly into the SON increased the concentration of AVP in plasma samples collected during simultaneous forced swimming. In an additional experiment the effect of the defined stressor exposure on the concentration of mRNA coding for nNOS within the SON was investigated by in situ hybridization. Forced swimming increased the expression of nNOS mRNA at two and four hours after onset of the stressor compared to untreated controls. Taken together, our results imply that NO within the SON does not contribute to the regulation of the secretory activity of HNS neurons during acute forced swimming. Increased nNOS mRNA in the SON after forced swimming and the increase in AVP release in the presence of exogenous NO under forced swimming points to a possible role of NO in the regulation of the HNS under repeated stressor exposure. PMID- 14752615 TI - Analysis of 6-hydroxy-2-aminocaproic acid (HACA) as a specific marker of protein oxidation: the use of N(O,S)-ethoxycarbonyl trifluoroethyl ester derivatives and gas chromatography/mass spectrometry. AB - An alteration of low density lipoprotein (LDL) apolipoprotein (apo) B-100 structure by direct oxidative modification is an important mechanism involved in atherogenesis. There is difficulty in quantifying this type of modification because a lack of specific assays. The use of N(O,S)-ethoxycarbonyl trifluoroethyl amino acid esters for a rapid and sensitive determination of 6 hydroxy-2-aminocaproic acid (HACA), a highly specific marker of metal catalyzed protein oxidation, by using standard gas chromatography/electron impact mass spectrometry, is discussed. The derivatives are formed by the unlabored reaction of amino acids with ethylchloroformate plus trifluoroethanol plus pyridine. Femtomole levels of HACA can be reproducible measured in different LDL preparations subjected to oxidative damage in the presence of iron or copper. HACA determination compares well with the measurement of carbonyl groups that are generally accepted as a nonspecific index of protein oxidation. Thus, the method could prove to be a sensitive assay for studying specific apoB-100 modification. PMID- 14752616 TI - Gender-related differences in carnosine, anserine and lysine content of murine skeletal muscle. AB - The aminoacyl-imidazole dipeptides carnosine (beta-alanyl-L-histidine) and anserine (beta-alanyl-1-methyl-histidine) are present in relatively high concentrations in excitable tissues, such as muscle and nervous tissue. In the present study we describe the existence of a marked sexual dimorphism of carnosine and anserine in skeletal muscles of CD1 mice. In adult animals the concentrations of anserine were higher than those of carnosine in all skeletal muscles studied, and the content of aminoacyl-imidazole dipeptides was remarkably higher in males than in females. Postnatal ontogenic studies and hormonal manipulations indicated that carnosine synthesis was up-regulated by testosterone whereas anserine synthesis increased with age. Regional variations in the concentrations of the dipeptides were observed in both sexes, skeletal muscles from hind legs having higher amounts of carnosine and anserine than those present in fore legs or in the pectoral region. The concentration of L-lysine in skeletal muscles also showed regional variations and a sexual dimorphic pattern with females having higher levels than males in all muscles studied. The results suggest that these differences may be related with the anabolic action of androgens on skeletal muscle. PMID- 14752617 TI - Effects of taurine supplementation on VDT work induced visual stress. AB - In order to evaluate the effects of dietary taurine supplementation on visual fatigue induced by visual display terminals (VDT) work, 25 male college students aged from 20 to 24 years who were not engaged in VDT work were selected to participate in the study. Volunteers were randomly assigned to either the taurine supplementation (n=13) or the placebo supplementation control group (n=12). Before and after 12 days of taurine (3 g/day) or placebo supplementation, two identical 2.5-hr VDT work tests were performed while recording the P100, N75 and N145 latencies and P100 amplitude of pattern visual evoked potential (PVEP) and the frequency of critical flicker fusion (CFF). Following 2.5-hr of VDT work, the P100 and N75 latencies of PVEP increased ( P<0.01) while the P100 amplitude decreased significantly ( P<0.01). The frequency of CFF also reduced significantly ( P<0.01). After 12 days of taurine supplementation, the reduction in P100 amplitude after VDT work alleviated significantly ( P<0.05). The results suggest that taurine supplementation alleviates visual fatigue induced by VDT work. PMID- 14752618 TI - Myocardial taurine, development and vulnerability to ischemia. AB - Depleting intracellular taurine in heart cells improves their resistance to ischemia and reperfusion injury. The aim of this work was to see whether physiologically low levels of endogenous taurine also reflect a reduced vulnerability of the myocardium to cardiac insults. The myocardial concentration of taurine was measured during different stages of development and compared with vulnerability to ischemia and reperfusion injury in the rat and in pediatric patients undergoing cardiac surgery.Rat hearts with relatively lower levels of taurine were significantly more resistant to an ischemic inult and there was a strong negative correlation between taurine content and recovery. Children's hearts had significantly lower taurine levels compared to infants' hearts which was consistent with their known increased resistance to an ischemic cardioplegic insult (Imura et al., 2001). This work shows that the changes in the concentration of myocardial taurine during development correlate with vulnerability to ischemia where low myocardial taurine is associated with improved recovery upon reperfusion. PMID- 14752619 TI - Plasma amino acids and sports injuries. AB - The aim of this study was to explore the relationship between changes in plasma amino acids and the incidence of sports injuries during a soccer season. Fourteen plasma amino acids were assayed at monthly intervals in 12 young soccer players during a whole soccer season. Based on the number and severity of injuries the soccer players were divided into an injury-prone and a non-injury-prone group. The mean plasma level of the amino acid glycine was significantly lower ( P=0.009) in the injury-prone group than the other group, while the mean plasma levels of tyrosine, tryptophan and lysine were higher in the injury-prone group during this period ( P<0.05). However there were no significant differences in the calculated plasma tryptophan and tyrosine/large neutral amino acids ratios. Significant linear time trends were observed for taurine, ornithine, lysine and the tryptophan/large neutral amino acids ratio.These results indicate that the plasma concentrations of glycine and to a lesser extent those of tyrosine, tryptophan and lysine may be promising peripheral markers for injury-proneness in young soccer players. Whether a role for glycine substitution will be indicative to reduce the occurrence of sports injuries will need to be investigated in future studies. PMID- 14752620 TI - Immunocytochemical study by two photon fluorescence microscopy of the distribution of GABA(A) receptor subunits in rat cerebellar granule cells in culture. AB - An immunocytochemical investigation of the expression of alpha(1), alpha(6), beta(2/3), gamma(2) and delta subunits was performed on rat cerebellum granule cells in culture by the two photon microscopy technique. The first four subunits appear to be expressed abundantly in these cells, whereas the delta one seems to be expressed at a lower level. Another major difference in the distribution of these subunits is that whereas alpha(6), beta(2/3) and gamma(2) appear only on plasma membranes alpha(1) and delta are present mainly in the cell bodies cytoplasm. Still another difference was found in that the presence of gamma(2) on neurites is "polarized", preferentially labelling neurites with the appearance of dendrites. The subunits alpha(6) and beta(2/3) appear to label all types of neurites, with beta(2/3) being by far the most heavily expressed subunit type. A final distinct characteristic is that alpha(6) and, even more, gamma(2) appear to accumulate in the cytoplasmic domains immediately below the cone of emergence of neurites. This suggests a conspicuous transport of such subunits from the site of synthesis in the cell body to the site of final expression in the neurites (dendrites and axon terminals). PMID- 14752621 TI - L-histidine is a beneficial adjuvant for antiepileptic drugs against maximal electroshock-induced seizures in mice. AB - Endogenous histamine has been reported to be involved in regulation of seizure susceptibility. Enhancement of histamine neurotransmission engendered by L histidine treatment produces anticonvulsant effects in experimental animals. The present study investigated the influence of L-histidine on the protective effects of carbamazepine and phenytoin against maximal electroshock-induced seizures in mice.L-Histidine, administered at the doses that did not influence the threshold for electroconvulsions (250-500 mg/kg), enhanced by nearly 30% the protective effects of carbamazepine against maximal electroshock-induced seizures. D Histidine (1000 mg/kg), an inactive isomer of histidine, was without any effect in this regard. L-Histidine (500 mg/kg) also augmented the protective effects of phenytoin. Importantly, the enhancement of the anticonvulsant effects of these antiepileptic drugs produced by L-histidine co-administration was not associated with augmentation of their unwanted effects on memory and motor performance. A pharmacokinetic interaction was also excluded since the free plasma levels of these antiepileptics remained unchanged in the presence of L-histidine. It may be suggested that L-histidine could serve as a beneficial adjuvant for selected antiepileptic drugs. PMID- 14752622 TI - Characteristics of taurine release induced by free radicals in mouse hippocampal slices. AB - The release of the inhibitory neuromodulator taurine in the hippocampus is markedly enhanced under various neural cell-damaging conditions, including ischemia and exposure to free radicals. The properties and regulation of the release evoked by a medium containing free radicals was investigated in hippocampal slices from adult (3-month-old) and developing (7-day-old) mice, using a superfusion system. The 'free radical damage' was induced by applying 0.01% H(2)O(2). The release of [(3)H]taurine was in both adult and developing hippocampus partly Ca(2+)-independent, mediated by Na(+)-dependent transporters and probably resulting from disruption of cell membranes and subsequent ion imbalance. The release in developing mice appeared to be more susceptible to regulation than that in adults, the stimulation by free radicals being in the latter already maximal. The release was reduced by adenosine A(1) receptor agonist R(-)N(6)-(2-phenylisopropyl)adenosine, which effect was, however, abolished by the antagonist 8-cyclopentyl-1,3-dipropylxanthine only in the immature hippocampus, indicating a receptor-mediated process. Moreover, the evoked taurine release in developing mice was potentiated by the ionotropic glutamate receptor agonists N-methyl-D-aspartate, kainate and 2-amino-3-hydroxy-5 methyl-4-isoxazolepropionate in a receptor-mediated manner, since the effects were abolished by their respective antagonists. The metabotropic glutamate receptors are of only minor significance in the release, the agonists of group I and II receptors slightly reducing the release. Furthermore, NO may also be involved in this release, the NO-generating compounds hydroxylamine and S-nitroso N-acetylpenicillamine being able to enhance the free-radical-evoked release. It seems that the free-radical-stimulated release, potentiated by ionotropic glutamate receptor activation and NO production, could constitute part of the neuroprotective properties of taurine, being important particularly in the developing hippocampus and hence preventing excitotoxicity. PMID- 14752623 TI - Evidence for expression of a single distinct form of mammalian cysteine dioxygenase. AB - Cysteine dioxygenase (CDO) plays a critical role in the regulation of cellular cysteine concentration. Because multiple forms of CDO ( approximately 23 kDa, approximately 25 kDa, and approximately 68 kDa) have been claimed based upon separation and detection using SDS-PAGE/western blotting (with antibodies demonstrated to immunoprecipitate CDO), we further investigated the possibility of more than one CDO isoform. Using either rabbit antibody raised against purified rat liver CDO or against purified recombinant his(6)-tagged CDO (r his(6)-CDO) and using 15% (wt/vol) polyacrylamide for the SDS-PAGE, we consistently detected the approximately 25 kDa band, but never detected a approximately 68 kDa band, in rat liver, kidney, lung and brain. Nondenatured gel electrophoresis of r-his(6)-CDO yielded a molecular mass estimate of 25.7 kDa and no evidence of dimerization. Mass spectrometry of r-his(6)-CDO yielded two peaks with molecular masses of 24.1 kDa and 24.3 kDa. Anion-exchange FPLC of r-his(6) CDO also gave two peaks, with the first containing CDO that was 7.5-times as active as the more anionic form that eluted second. When the two peaks recovered from FPLC were run on SDS/PAGE, the first (more active) CDO fraction yielded two bands (perhaps as an artifact of SDS/PAGE), whereas the second (less active) CDO fraction yielded only the approximately 23 kDa band. We conclude that the physiologically active form of CDO is the approximately 25 kDa (i.e., 23.5 kDa based on mass spectrometry) monomer and that this active form is probably derived by post-translational modification of the 23 kDa gene product. PMID- 14752624 TI - Failed back surgery syndrome: the role of symptomatic segmental single-level instability after lumbar microdiscectomy. AB - Segmental instability represents one of several different factors that may cause or contribute to the failed back surgery syndrome after lumbar microdiscectomy. As segmental lumbar instability poses diagnostic problems by lack of clear radiological and clinical criteria, only little is known about the occurrence of this phenomenon following primary microdiscectomy. Retrospectively, the records of 2,353 patients were reviewed according to postoperative symptomatic segmental single-level instability after lumbar microdiscectomy between 1989 and 1997. Progressive neurological deficits increased (mean of 24 months; SD: 12, range 1 70) after the initial surgical procedure in 12 patients. The mean age of the four men and eight women was 43 years (SD: 6, range 40-77). The main symptoms and signs of secondary neurological deterioration were radicular pain in 9 of 12 patients, increased motor weakness in 6 of 12 patients and sensory deficits in 4 of 12 patients. All 12 symptomatic patients had radiological evidence of segmental changes correlating with the clinical symptoms and signs. All but one patient showed a decrease in the disc height greater than 30% at the time of posterior spondylodesis compared with the preoperative images before lumbar microdiscectomy. All patients underwent secondary laminectomy and posterior lumbar sponylodesis. Postoperatively, pain improved in 8 of 9 patients, motor weakness in 3 of 6 patients, and sensory deficits in 2 of 4 patients. During the follow-up period of 72+/-7 months, one patient required a third operation to alleviate spinal stenosis at the upper end of the laminectomy. Patients with secondary segmental instability following microdiscectomy were mainly in their 40s. Postoperative narrowing of the intervertebral space following lumbar microdiscectomy is correlated to the degree of intervertebral disc resection. It can therefore be concluded that (1) patients in their 40s are prone to postoperative narrowing of the intervertebral disc space and hence subsequent intervertebral instability and (2) that a small extent of intervertebral disc resection and preservation of the "segmental frame" may be beneficial in those patients. The present study demonstrated for the first time that the degree of extensive operative techniques in microdiscectomy increased the risk of subsequent segmental instability. In addition, narrowing of the intervertebral space of more than 30% represents a clear radiological sign of segmental instability. PMID- 14752625 TI - Measuring mucosal damage induced by cytotoxic therapy. AB - We scored oral mucositis and gut toxicity and measured sugar permeability testing among 56 recipients of a haematopoietic stem cell transplant (HSCT) given myeloablative conditioning with idarubicin, cyclophosphamide and TBI, and a group of 18 patients given cytotoxic chemotherapy for newly diagnosed acute myeloid leukaemia (AML) or myelodysplastic syndrome (MDS). Gut integrity was already disturbed in the AML/MDS group as measured by the lactulose/rhamnose ratio (L/R ratio=0.09) before therapy and was severely perturbed (L/R ratio >0.13) for a month after HSCT. Oral mucositis and to a lesser extent gut toxicity was only significantly correlated with disturbed permeability in the transplant group. The data suggest that sugar permeability, oral mucositis and gut toxicity measure different features of mucosal damage after intensive cytotoxic therapy. PMID- 14752626 TI - Outcome of laparoscopic colorectal resection. AB - BACKGROUND: The aim of this study was to assess the feasibility and safety of laparoscopic surgery for colorectal diseases. METHODS: A retrospective review was undertaken of all patients undergoing a laparoscopic colorectal procedure (LCP) for large bowel disease. All operations were performed by a single experienced team. Patients were divided chronologically into three consecutive groups (G1, G2, and G3). Data collection included the incidence and cause of both "proper" and "mandatory" conversions to laparotomy, the incidence and type of early and late postoperative complications, incidence of operative mortality, and the length of hospital stay. The incidences of conversion to laparotomy and of early and late postoperative complications were also determined as related to diagnosis, type of LCP attempted, and chronological group. RESULTS: Between January 1996 and December 2001, a total of 108 patients (49 men and 59 women) with a mean age of 65.1 years underwent an LCP for colorectal disease. Proper conversion to open surgery was necessary in five patients (4.6%), whereas a mandatory conversion was needed in 10 with patients advanced cancer (9.2%). The overall morbidity rate was 11.9%. There were no anastomotic leaks. In two patients (1.85%) developed a complication requiring reoperation. Postoperative mortality was nil. Mean postoperative hospital stay was 7.2 days. The rates of conversion and of early and late complications decreased through the three chronological periods. No trocar site recurrences were observed in the cancer patients. CONCLUSION: Laparoscopic colorectal surgery performed in experienced centers is safe; the observed morbidity and mortality rates are low and acceptable and compare favorably to those observed after standard open surgery. PMID- 14752627 TI - Favorable early results of gastric banding for morbid obesity: the American experience. AB - BACKGROUND: In 2001 a new device for surgical weight loss was approved by the Food and Drug Administration (Lap-Band, Inamed Health). We describe initial results of laparoscopic gastric banding for morbid obesity in two American academic centers. METHODS: Prospective data was collected on consecutive morbidly obese patients undergoing laparoscopic adjustable gastric banding, and evaluated retrospectively. RESULTS: Four hundred forty-five consecutive patients underwent Lap-Band from May 2001 through December 2002. The 103 men and 341 women had an average age of 42.1 years (range 17-72 years) and an average body mass index (BMI) of 49.6 kg/m2 (range 35.2-92.2 kg/m2). One operation required conversion to laparotomy due to bleeding; the rest were completed laparoscopically. Mean length of stay was 1.1 days (range 1-10 days). There was one death. Additional complications included band slippage in 14 patients (3.1%), gastric obstruction without slip in 12 (2.7%), port migration in 2 (0.4%), tubing disconnections in 3 (0.7%), and port infection in 5 (1.1%). Two bands (0.4%) were removed due to intraabdominal abscess 2 months after placement. There was one band erosion (0.2%) and no clinically significant esophageal dilation. Ninety-nine patients have 1-year follow-up and have lost an average of 44.3% excess body weight. CONCLUSION: Laparoscopic gastric banding has much to offer the morbidly obese. We present data showing weight loss rivaling gastric bypass and acceptably low complications. These results parallel success with this device outside America. PMID- 14752628 TI - Laparoscopic resection of the pancreas: a feasibility study of the short-term outcome. AB - BACKGROUND: Laparoscopic resection is not an established treatment for tumors of the pancreas. We report our preliminary experience with this innovative approach to pancreatic disease. METHODS: Thirty two patients with pancreatic disease were included in the study on an intention-to-treat basis. The preoperative indications for surgery were as follows: neuroendocrine tumors ( n=13), unspecified tumors ( n=11), cysts ( n=2), idiopathic thrombocytopenic purpura with ectopic spleen ( n=2), annular pancreas ( n=1), trauma ( n=1), aneurysm of the splenic artery ( n=1), and adenocarcinoma ( n=1). RESULTS: Enucleations ( n=7) and distal pancreatectomy with ( n=12) and without splenectomy ( n=5) were performed. Three patients underwent laparoscopic exploration only. Four procedures (13%) were converted to an open technique. One resection was converted to a hand-assisted procedure. The mortality rate for patients undergoing laparoscopic resection was 8.3% (two of 24). Complications occurred after resection in nine of 24 procedures (38%). The median hospital stay was 5.5 days (range, 2-22). Postoperatively, opioid medication was given for a median of 2 days (range, 0-13). CONCLUSION: Resection of the pancreas can be performed safely via the laparoscopic approach with all the potential benefits to the patients of minimally invasive surgery. PMID- 14752629 TI - The benefits of stereoscopic vision in robotic-assisted performance on bench models. AB - BACKGROUND: Previous studies have failed to establish clear advantages for the use of stereoscopic visualization systems in minimal-access surgery. The aim of this study was to objectively assess whether stereoscopic visualization improves performance on bench models using the da Vinci robotic system. METHODS: Eleven surgeons carried out a series of four tasks. Positional data streamed from the da Vinci system was analyzed by means of a previously validated custom-designed software-package. An independent blinded observer scored errors. Statistical analysis included the Wilcoxon signed rank test. A p < 0.05 was deemed significant. RESULTS: We found significant improvements in all tasks and for all parameters (p < 0.05). In addition, a significantly lower number of errors was scored using the stereoscopic mode as compared to the standard two-dimensional image (p < 0.001). CONCLUSION: Robotic-assisted performance on bench models is more efficient and accurate using stereoscopic visualization. PMID- 14752630 TI - Intraoperative endoscopic retrograde cholangiopancreatography (ERCP) to remove common bile duct stones during routine laparoscopic cholecystectomy does not prolong hospitalization: a 2-year experience. AB - BACKGROUND: There is still some controversy regarding the optimal timing and best method for the removal of common bile duct stones (CBDS). Intraoperative endoscopic retrograde cholangiopancreaticography (IO-ERCP) is an alternative method that should be considered for this procedure. The aim of our study was to investigate the clinical outcome of a single-step procedure (IO-ERCP) to remove CBDS, thereby combining two existing high-volume clinical modalities-i.e., laparoscopic cholecystectomy (LC) and ERCP. METHODS: Between January 2000 and December 2001, 674 patients, 192 male and 482 female, underwent cholecystectomy at our hospital. There were 612 LC (90.8%), 37 converted procedures (5.5%), and 25 open operations (3.7%). In 592 of the patients, (87.8%) intraoperative cholangiography (IOC) was performed. In 34 (5.7%) of those who had and IOC, an IO ERCP was performed. While the surgeon waited for the endoscopist, care was taken to introduce a thin guidewire through the lOC catheter and pass it through the sphincter of Oddi, out into the duodenum. This complementary procedure greatly facilitated the subsequent cannulation of the bile ducts. RESULTS: The cannulation frequency of the CBD was 100%. Common bile duct stones were successfully extracted in 93.5%. Endoscopic sphincterotomy (EST), followed by the insertion of a plastic endoprosthesis, was performed in two patients with remaining stones. The CBD of these two patients was cleared by postoperative ERCP. None of the patients developed postoperative pancreatitis. The operating time was prolonged as compared with the time for LC (192 vs 110 mins; p < 0.05). The length of hospitalization for IO-ERCP patients did not differ from that for patients undergoing cholecystectomy alone (2.6 vs 2.1. days; NS). CONCLUSIONS: The study suggests that elective IO-ERCP is a safe and efficient method for removing CBDS that has a low risk of inducing postoperative pancreatitis and does not prolong postoperative hospitalization. This technique enables perioperative extraction of CBDS without open or laparoscopic surgical exploration of the CBD and can be used safely in a routine clinical setting. PMID- 14752631 TI - Laparoscopic management of common bile duct stones. AB - BACKGROUND: While laparoscopic cholecystectomy is widely accepted for therapy of cholecystolithiasis, controversy still exists concerning the management of common bile duct stones. Besides preoperative endoscopic papillotomy followed by laparoscopic cholecystectomy and open common bile duct surgery, management of common bile duct stones can be conducted by laparoscopy, if respective experience is available. METHOD: During laparoscopic cholecystectomy a cholangiography via the cystic duct is routinely performed. If bile duct stones are detected they are retrieved via the cystic duct or via incision of the common bile duct by insertion of a Fogarty catheter or Dormia basket. Exclusion criteria against simultaneous laparoscopic management include suspicion of malignancy, severe pancreatitis, or cholangitis. RESULTS: From November 1991 to March 2002, 200 patients primarily underwent laparoscopic therapy of bile duct stones. Retrieval was performed via cystic duct and common bile duct incision in 115 and 85 cases, respectively. Complete removal was achieved in 91%; complication rate and mortality was 7% and 0.5%, respectively. During the same period primary endoscopic papillotomy was necessary in 40 patients because of the above contraindications. CONCLUSIONS: When correct indications and surgical expertise are observed, simultaneous laparoscopic management of common bile duct stones represents a safe and minimally invasive alternative to a two-procedure approach. PMID- 14752632 TI - Endoscopic implantation and patency evaluation of lumboperitoneal shunt: an innovative technique. AB - BACKGROUND: The placement of the lumboperitoneal (LP) shunt tube used in the management of idiopathic intracranial hypertension (IIH) and the evaluation of its patency necessitate an abdominal surgical incision. This procedure can now be done using a laparoscopic-assisted technique. This study was designed to evaluate the usefulness of this technique in treating patients with IIH in whom visual loss was progressive in spite of aggressive medical management, as well as for the evaluation of the function of the shunt tube after its placement. METHODS: Seventeen patients aged between 21 and 45 years (mean, 31) were included in the study. They were divided into two groups. Laparoscopy was used in the first group of 11 patients for primary placement of the peritoneal portion of shunt catheter in the right subphrenic recess. It was used in the second group, which consisted of six patients who had recurrence of symptoms after surgical LP shunt placement, for the evaluation of shunt patency and position inside the peritoneal cavity and for the repositioning of the displaced shunt, as needed. RESULTS: In the first group (n = 11), visual symptomatology was improved in 10 of 11 patients and became stable in the remaining one. In the second group (n = 6), two of six patients had a patent tube in a proper position; three had complete intraperitoneal migration of the shunt tubes, which were repositioned using a laparoscopic-assisted technique; and the last patient had occlusion of the peritoneal side of the shunt by omental adhesions that had been liberated by the laparoscopy. No complications related to laparoscopy were recorded in this series. CONCLUSION: This procedure was associated with better functional results, less postoperative pain and discomfort, a shorter hospital stay, an earlier return to normal activities, and cosmetic acceptability . PMID- 14752633 TI - Laparoscopic virtual reality and box trainers: is one superior to the other? AB - BACKGROUND: Virtual reality (VR) simulators now have the potential to replace traditional methods of laparoscopic training. The aim of this study was to compare the VR simulator with the classical box trainer and determine whether one has advantages over the other. METHODS: Twenty four novices were tested to determine their baseline laparoscopic skills and then randomized into the following three group: LapSim, box trainer, and no training (control). After 3 weekly training sessions lasting 30-min each, all subjects were reassessed. Assessment included motion analysis and error scores. Nonparametric tests were applied, and p < 0.05 was deemed significant. RESULTS: Both trained groups made significant improvements in all parameters measured ( p < 0.05). Compared to the controls, the box trainer group performed significantly better on most of the parameters, whereas the LapSim group performed significantly better on some parameters. There were no significant differences between the LapSim and box trainer groups. CONCLUSIONS: LapSim is effective in teaching skills that are transferable to a real laparoscopic task. However, there appear to be no substantial advantages of one system over the other. PMID- 14752635 TI - Comparison of high- vs low-dose 5-aminolevulinic acid for photodynamic therapy of Barrett's esophagus. AB - BACKGROUND: Barrett's esophagus is the major risk factor for esophageal adenocarcinoma, the incidence of which is increasing rapidly in the Western world. Aminolevulinic acid for photodynamic therapy (ALA-PDT) is effective in the treatment of Barrett's esophagus, but controversy exists regarding optimum ALA dosage. The aim of this study was to establish the optimum dosage regime for ALA PDT for Barrett's esophagus. METHODS: Twenty-five patients with Barrett's esophagus were randomized to receive 30 (low-dose) or 60 (high-dose) mg/kg oral ALA at 4 or 6 h or 30 mg/kg in two fractions 4 and 6 h before PDT. PDT was standardized using red (635 nm) light. Biopsy specimens were taken for protoporphyrin IX (PpIX) quantification. Endoscopy was repeated 4 weeks later. RESULTS: All patients showed a macroscopic response, with squamous re epithelialization. This response was greatest in the 30 mg/kg and fractionated ALA groups. There was no significant difference in response between dosing 4 or 6 h prior to PDT. Tissue levels of PpIX were similar for all dosage groups and were not predictive of clinical response. Side effects were more common with the higher dose of ALA. CONCLUSION: Low-dose ALA-PDT appears to be a safe protocol for the ablation of Barrett's esophagus. PMID- 14752634 TI - Qualitative and quantitative analysis of the learning curve of a simulated surgical task on the da Vinci system. AB - BACKGROUND: Robotic telemanipulation systems provide solutions to the problems of less dexterity and visual constraints of minimally invasive surgery (MIS). However, their influence over surgeons' dexterity and learning curve needs to be assessed. We present motion analysis as an objective method to measure performance and learning progress.METHODS. Thirteen surgeons completed five synthetic small bowel anastomoses using the da Vinci system. Objective Structured Assessment of Technical Skills (OSATS) allowed qualitative analysis. Quantitative analysis used API software of the system to retrieve real-time robotic signal data of time, path length, and number of movements. Wilcoxon signed ranks test was used for statistical analysis. A p value <0.05 was considered significant.RESULTS. OSATS global scores were 18.6 points for the first attempt and 26 for the fifth attempt ( p < 0.02, Cronbach's alpha = 0.894). Paired data of motion analysis for attempts 1 vs 5 showed significant change: time taken 3507 sec and 2287 sec ( p < 0.008), total number of movements 2411 and 1387 ( p = 0.01), total path length 21,630 cm and 13,941 cm ( p = 0.01).CONCLUSIONS. A rapid learning curve to a competent level using the da Vinci system is possible aided by the system's intuitive motion. Motion analysis is a useful tool to measure performance in the da Vinci system compared to OSATS and time alone. PMID- 14752636 TI - First experience of endoscopic implantation of tissue expanders in plastic and reconstructive surgery. AB - BACKGROUND: Over the last 15 years, tissue expansion has been used in the treatment of 324 patients at the A. V. Vishnevsky Institute of Surgery. The principal drawbacks of the traditional technique for tissue expansion are the prolonged time needed to complete the process and a high rate of complications. With the traditional technique, the wound created by the implantation of the expander inhibits the expansion of the tissue until healing takes place, which requires 10-14 days. However, attempts to reduce of the length of the incision have been limited by the necessity to ensure good hemostasis in the expander pocket. We describe our initial experience with the endoscopic insertion of tissue expanders. METHODS: We performed the endoscopic implantation of 20 tissue expanders in nine patients. Six patients had postburn scar deformities, one had congenital microtia, one had a capillary malformation, and one had a capillary malformation combined with postburn scar deformity. Tissue expanders were inserted in the head, face, neck, chest wall, scapular, shoulder, forearm, and calf regions. The endoscopic technique was specially adapted according to the features of the different anatomic sites. Endoscopy enabled creation of the expander pocket, with good control of hemostasis, through incisions < or =1 cm. The orientation of these incisions was parallel to forces of tension. Therefore, full expansion began immediately after placement of the expander, without any risk of postoperative wound dehiscence. The expanders were fully inflated intraoperatively accordingly to the principals followed in the traditional technique of implantation for inflation over a 3-week period. Further inflations were carried out regularly either daily or once every other day. RESULTS: The average time required for the tissue expansion was 34 days, which was less than half the time needed with the traditional technique. All flaps remained extant after reconstruction with the expanded tissues. There were no complications. CONCLUSION: The advantages of the endoscopic implantation of tissue expanders are a reduction in expansion time, a shorter hospital stay, less patient discomfort, and the prevention of complications (hematoma, seroma, infection, wound dehiscence). PMID- 14752637 TI - New model for skills assessment and training progress in minimally invasive surgery. AB - BACKGROUND: With the spread of minimally invasive surgery and training in this field, development of metrics for skills assessment and training progress has become increasingly important. Our approach was to use the tracking of motion for the definition of objectives metrics. METHODS: We have developed an inanimate model and tracked the 3D coordinates of the instrument tips with an ultrasound system. Besides already validated parameters (time, error time, and distance efficiency ratio) we examined the transit and the speed profile for their evidentiary power. Performances of experts (who have already performed >100 laparoscopic operations) and novices (<20 laparoscopic operations) were evaluated. RESULT: The standardized time, the error time as a precision indicator, and the transit profile parameter for spatial perception could significantly ( p < 0.05) distinguish between experts and novices. Furthermore, these parameters and the distance efficiency ratio improved significantly during a training course in laparoscopic surgery. CONCLUSION: Our model showed changes of the mentioned parameters with experience. According to our results, it can be used for skills assessment and as a training progress measurement system. We propose transit profile as an additional important parameter for assessment. PMID- 14752638 TI - Results of thoracoscopic pleural abrasion for primary spontaneous pneumothorax. AB - BACKGROUND: Several video-assisted techniques have been used to treat primary spontaneous pneumothorax (PSP). The aim of this study was to evaluate the results of thoracoscopic pleural abrasion for PSP. METHODS: From 1991 to 2003, 185 consecutive patients, 143 male and 42 female, aged 15 to 60 years (average 31.6) underwent thoracoscopic pleural abrasion for PSP. The indications for surgery were as follows: a first episode with persistent air leak in 33 patients (17.9%), a recurrent ipsilateral pneumothorax in 122 patients (65.9%), a previous contralateral pneumothorax in 23 patients (12.4%), and recurrence after surgical treatment in seven patients (3.8%). Bullae were resected in 163 patients (88.1%). Mechanical pleural abrasion was performed in all cases. RESULTS: There were no deaths. Intraoperative hemorrhage occurred in three patients. It was controlled via thoracotomy in one patient and via thoracoscopy in two patients. The postoperative complication rate was 8.1% (15/185). Complications included prolonged air leak in eight patients (4.3%), pleural effusion in two (1.1%), extrapleural hematoma in one (0.5%), chest wall infection in one (0.5%), atelectasis in one (0.5%), and hemorrhage in two (1.1%). Postoperative hospital stay ranged between 2 and 17 days (mean, 5). Mean duration of drainage was 3.8 days (range; 1-16). Postoperatively, 111 patients were contacted, with a mean follow-up of 36.5 months. Four of them had a recurrence (3.6%) that did not require reoperation. CONCLUSION: Thoracoscopic pleural abrasion associated with bullae resection is a safe and efficient treatment for PSP. Results remain stable in the long term. PMID- 14752639 TI - Antibiotic prophylaxis in elective laparoscopic cholecystectomy. Lack of need or lack of evidence? AB - BACKGROUND: The need to administer antibiotic prophylaxis (ABP) during laparoscopic cholecystectomy (LC) is still a matter of significant controversy. The purpose of this study was to resolve this issue by performing a meta-analysis of the available randomized controlled trials (RCT) on this topic. METHODS: Papers identified via a systematic literature search were evaluated according to standard criteria. Data regarding the patient sample, study methods, and outcomes were abstracted and summarized across studies. The outcome measures were the rates of all perioperative infections, the rates of surgical site infections, and the rates of infections at other sites. Results were examined for 974 patients randomized to ABP or placebo prior to LC in six RCT published from 1997 to 2001. RESULTS: The cumulative rates of all infections were 2.8% in the ABP group and 4.4% in the placebo group. The pooled odds ratio (OR) (95% confidence interval [CI]) was 0.69 (0.34-1.43; p = 0.32). The cumulative rates of surgical site infections were 2.1% in the ABP group and 2.9% in the placebo group. The pooled OR (95% CI) was 0.82 (0.36-1.86; p = 0.63). The cumulative rates of infections at other sites were 0.7% in the ABP group and 1.5% in the placebo group. Pooled OR (95% CI) was 0.82 (0.18-1.90; p = 0.37). No significant heterogeneity was found in any data pooling. CONCLUSIONS: Based on the available evidence, there appears to be no need to administer routine ABP to low-risk patients during LC. However, the number of patients enrolled to date into RCT is insufficient to avoid a type II error. A large and well-designed trial is urgently needed to find a conclusive answer to this question. PMID- 14752640 TI - Effects of pneumoperitoneum created through CO2 insufflation and parameters of mechanical ventilation (PEEP application) on systemic dissemination of intraabdominal infections. AB - BACKGROUND: To examine whether CO2 pneumoperitoneum and positive end expiratory pressure (PEEP) in mechanical ventilation affect the systemic spread of intraabdominal infection. METHODS: Sprague-Dawley male rats weighing 200-300 g were allocated to three groups of 12 animals in each. All rats received mechanical ventilation under general anesthesia. An intraabdominal infection model was established by injecting with 1 ml of Escherichia coli (10(9) CFU/mL) intraperitoneally. Half of the animals in each group were exposed to PEEP (10 cmH2O). CO2 pneumoperitoneum at 13 mmHg was applied to the rats in group 1. Group 2 rats underwent laparotomy. Group 3 served as controls. In addition, TNF-alpha serum levels were measured at baseline and 3 h. A peritoneal specimen for histopathological examination were obtained after the rats were killed at the end of 3 h. For the assessment of data, descriptive statistical methods (mean, standard deviation) as well as Friedman test for repeated measurements in multiple groups, Kruskal-Wallis test for intergroup comparisons, Dunn's multiple comparison test for subgroup comparisons, Mann-Whitney U test for comparisons between paired groups, chi-square and Fisher's exact test for comparison of qualitative data, and McNemar's test for assessment of changes in group variables over time were used. The results were considered statistically significant if probability (p) values were <0.05. RESULTS: Grades of peritonitis in group 1 and 2 were seen to differ nonsignificantly. In group 1, baseline blood cultures were not included in the assessment between the subgroups that received PEEP or not, as there was no growth in any of the subgroups. No significant difference was detected between growth in blood cultures at 1, 2, and 3 h ( p > 0.05). Application of PEEP in subgroups did not alter the blood culture results ( p > 0.05). Significant differences were seen between the initial and final TNF-alpha values of groups (KW: 18.94, p < 0.0001). The values in control group were observed to be significantly lower than those in groups 1 and 2 ( p < 0.01, p < 0.001). Bacteremia and systemic spread of the intraabdominal infection did appear to be different according to the PEEP application. After the assessments of ventilation parameters in our study, significant reductions in pH and HCO3 levels were detected in group 1 as a result of pneumoperitoneum, which was consistent with the literature. There is a significant difference between pH values at baseline and at the end of 1 h because of pneumoperitoneum (Fr: 10.01, p < 0.05). PEEP application in subgroups did not create significant differences in terms of respiratory parameters ( p < 0.01). CONCLUSION: No difference was found between the applications of CO2 pneumoperitoneum and laparotomy with regard to bacteremia and infection-induced peritonitis. It was determined that pneumoperitoneum along with PEEP application had neither a positive nor a negative impact on intraabdominal infection. PMID- 14752641 TI - Outcome of laparoscopic Nissen-Rossetti fundoplication in children with gastroesophageal reflux disease and supraesophageal symptoms. AB - BACKGROUND: The role of surgery is debated for children with gastroesophageal reflux disease (GERD), particularly when they show atypical symptoms. This study was designed to evaluate the safety and outcome of laparoscopic Nissen-Rossetti fundoplication performed in a selected population of children with gastroesophageal reflux and atypical supraesophageal symptoms. METHODS: This prospective study included 595 patients younger than 14 years with GERD who reported recurrent respiratory symptoms and had no benefit from standard medical treatment. Surgery was performed for 48 patients with anatomic anomalies, life threatening events, or respiratory complications after ineffective medical treatment. The subjective and objective outcomes were evaluated. RESULTS: No major intraoperative complications were experienced, and there was no recurrence of gastroesophageal reflux during a postoperative follow-up period of 12 months. The parents' final subjective evaluation of the outcomes 12 months after surgery was positive in 44 cases and negative in 4 cases. CONCLUSIONS: Children with difficult-to-treat chronic respiratory symptoms must be evaluated for GERD, even if the need for surgery is low (8%), because complete eradication of reflux is mandatory. Radical treatment of GERD allows the pulmonologist to perform correct respiratory treatment and to prevent the development chronic and life-threatening complications. PMID- 14752644 TI - Long-term retention of endoscopically placed hydrogel prostheses at the lower esophageal sphincter in pigs. AB - BACKGROUND: Using flexible endoscopic methods, hydrogel prostheses can be safely placed, retained for long periods of time, and removed from the esophageal submucosa of pigs. This new technique may have future applications in the treatment of gastroesophageal reflux disease in selected situations. METHODS: In a controlled, prospective trial, farm pigs ( n = 28) or Sinclair mini-swine ( n = 18) underwent sequential placements of 2-13 pliable, radio-opaque hydrogel prostheses into the submucosa of the esophagus during a single endoscopy session. A novel endoscope-overtube device was used. Followup endoscopy and/or fluoroscopy sessions were video-recorded at 6-week, 12-week, 6-month, and yearly intervals. The endoscopic removal of hydrogels was tested in vivo. Necropsy specimens were inspected for signs of chronic inflammation. RESULTS: Overall, 98% of delivery attempts were successful (288/293). Only three hydrogels were lost after 6 weeks. Short-term animals ( n = 36) retained 88% of hydrogels for periods up to 6 months. Intact hydrogels ( n = 12) were easily removed from four animals at endoscopy. Long-term subjects had two or three hydrogels per animal, using either a beveled-needle device (six pigs) or a trocar design (four pigs). Trocar design: nine of nine prostheses retained at 3 years (100%). Needle design: nine of 10 prostheses retained at 3 years (90%). No significant adverse event occurred. Growth curves were similar between groups. All hydrogels remained pliable to gross inspection upon in vivo removal or necropsy. Pathology showed minimal fibrosis and no chronic inflammation. CONCLUSIONS: A novel endoscopic overtube device allows for the successful delivery of multiple hydrogel prostheses with acceptable safety and long term retention rates. These hydrogel prostheses can also be easily removed from the submucosal space of the esophagus. PMID- 14752642 TI - Hematological long-term results of laparoscopic splenectomy for patients with idiopathic thrombocytopenic purpura: a case control study. AB - BACKGROUND: Laparoscopic splenectomy (LS) for idiopathic thrombocytopenic purpura (ITP) appears, when compared to open splenectomy (OS), associated with immediate important advantages. However, in a number of patients splenectomy does not lead to an adequate response, or after initial adequate response a relapse occurs after some time. A relapse may be associated to the presence of accessory spleens and splenosis. The purpose of this study was to compare the operative outcome and the hematological results on the long term of a series of LS with a historic series of OS for the treatment of ITP. METHODS: A retrospective review was done of 50 consecutive patients who underwent LS for ITP. Patient characteristics, outcome of surgery, and hematological results were compared to a historical group of patients who underwent conventional splenectomy for ITP (n = 31). Response to splenectomy was defined in three groups: complete remission, partial remission, and no response. Grouping was based on hematological data. RESULTS: Concerning operative outcome and postoperative complications, there was a significant difference in favor of LS. Moreover, the hematological outcome of both groups showed no differences after a median period of 66 months (OS) and 35 months (LS), respectively. CONCLUSIONS: Hematological results after laparoscopic splenectomy for ITP are comparable to those after open splenectomy in both the short and the long term. PMID- 14752645 TI - Laparoscopic approach for early gastric cancer within a paraesophageal hiatal hernia. AB - We present a case of early gastric cancer located in gastric volvulus associated with paraesophageal hiatal hernia. Two lesions of EGC were diagnosed in the distal third of the stomach, most of which had herniated into the left chest through a large hiatal defect in an organoaxial fashion. Routinely, laparoscopic assisted distal gastrectomy (LADG) is our preferred approach for EGC, and the presence of hiatal hernia in this case did not alter our approach. Laparoscopic repair of hiatal hernia was performed successfully followed by LADG. A review of the literature supports a minimally invasive approach for both procedures and shows it to be safe, effective, and technically feasible. Further, LADG is shown to be oncologically adequate in terms of tumor margins and lymph node dissection, but its relevance to long-term disease-free survival still needs to be studied in well-designed prospective trials. PMID- 14752646 TI - Magnitude of abdominal incision affects the duration of postoperative ileus in rats. AB - BACKGROUND: The pathogenesis of reduced postoperative ileus (POI) in laparoscopic gastrointestinal (GI) surgery still remains controversial. The aim of this study was to investigate the effect of surgical incision on postoperative ileus. METHODS: The effects of length, depth, and site of the incision on GI transit were compared using the geometric center of 51Cr in rats. The inhibitory mechanism of abdominal incision on GI transit also was studied. RESULTS: The findings showed that 5 cm of abdominal skin and the 5-cm back muscle incision had no significant effect on GI transit. However, the 5-cm abdominal muscle-fascia incision and a 5-cm laparotomy significantly delayed GI transit. Gastrointestinal transit after a 5-cm laparotomy was significantly delayed, as compared with that of a 1-cm laparotomy regardless whether intestinal manipulation was performed or not. Guanethidine and yohimbine, but not propranolol, significantly improved the impaired GI transit after a 5-cm laparotomy. CONCLUSIONS: The results suggest that the longer and deeper abdominal incision more profoundly inhibits GI transit. The inhibitory effect of abdominal incision is mediated via the activation of the somatosympathetic reflex and alpha-2 adrenoceptors. PMID- 14752647 TI - Investigating a possible cause of mesh migration during totally extraperitoneal (TEP) repair. AB - BACKGROUND: In experienced hands, laparoscopic inguinal hernia repair has a low rate of recurrence, but it still can recur, and a number of reasons for this have been identified. In published studies, the majority of such cases seem to result from inadequate dissection leading to missed hernias or suboptimal mesh placement. But even with adequate dissection and proper placement of a sufficiently large mesh, recurrence sometimes happens. A number of investigators have cited mesh migration or dislocation as a possible cause, and this study examined how hip flexion affects the position of newly placed meshes and staples in totally extraperitoneal (TEP) repair of inguinal hernia. METHODS: After completion of the dissection and reduction of discovered hernias, a 15 x 15-cm polypropylene mesh was placed either unilaterally or bilaterally, as indicated. The preperitoneal space then was desufflated. The operating table, in an extended -20 degrees position during surgery, was placed in a 90 degrees position for approximately 15 s. After reinsufflation, the possibility of mesh migration and folding was investigated. Finally, the mesh was stapled, the table again extended and flexed, and the possibility of mesh migration and staple dislodgement investigated once more. RESULTS: The mesh did not migrate or become displaced from any potential hernia area, nor did any of the staples become dislodged. CONCLUSIONS: Concern about mesh migration attributable to patients sitting up immediately after surgery appears to be unfounded, at least according to the findings for the current, small, simulated study group. PMID- 14752648 TI - Experience with routine intraabdominal cultures during laparoscopic gastric bypass with implications for antibiotic prophylaxis. AB - BACKGROUND: Techniques for laparoscopic Roux-en-Y gastric bypass that do not use bowel cross-clamping raise a question of increased risk for infectious complications. However, to the authors' knowledge, no studies have recorded routine intraoperative peritoneal cultures. This article reports the authors' experience with routine peritoneal cultures during laparoscopic Roux-en-Y gastric bypass and the role of antibiotic prophylaxis. METHODS: From January 2000 to March 2000, 66 consecutive patients undergoing a laparoscopically divided proximal Roux-en-Y gastric bypass had peritoneal fluid aspirated for routine culture. No mechanical or oral antibiotic bowel preparation was used. All the patients received preoperative intravenous antibiotic prophylaxis with Levoquin 500 mg and Flagyl 500 mg. Peritoneal fluid was aspirated from the left gutter near the site of the enteroenterostomy before irrigation with 1,000 ml of normal saline containing 50,000 U of bacitracin and 1 of kanamycin. RESULTS: The follow up period averaged 9 months for 100% of patients. For 15 patients (22.7%), the culture results were positive. The 22 organisms cultured involved 15 streptococcus species, 4 anaerobes, 2 staphylococcus species, and 1 enterobacter. None of the patients experienced a clinical infection or required an extension of antibiotics beyond the first 24 h. CONCLUSIONS: This study demonstrated frequent peritoneal contamination during laparoscopic gastric bypass. Prophylactic intravenous antibiotics and antibiotic irrigation may have reduced the risk of clinically significant infections in this small uncontrolled series. PMID- 14752649 TI - Totally extraperitoneal (TEP) hernia repair after an original TEPIs it safe, and is it even possible? AB - BACKGROUND: There are only scant published reports of totally extraperitoneal (TEP) repair of recurrence after a primary TEP procedure. Furthermore, at least two authors have made the statement that such an operation is virtually impossible. METHODS: We have been performing TEP repair of recurrence after TEP since we 1996, and here we present a retrospective review of our experience with the procedure. We employ a method not varying greatly from the standard TEP done for primary hernia. RESULTS: All cases were started laparoscopically, and only one of 20 had to be converted to open. Of these cases, 12 were for same-side recurrence and eight for a contralateral new hernia. With a follow-up of 28-74 months, there have been no fatalities, no complications, and no re-recurrence. CONCLUSION: We have found that TEP repair of recurrent inguinal hernia after a primary TEP repair is entirely feasible technically as well as entirely safe. PMID- 14752650 TI - Laparoscopic incisional hernia repair in a porcine model: what do transfixion sutures add? AB - BACKGROUND: The aim of this study was to evaluate the need for transfixion sutures during laparoscopic ventral hernia repair with mesh. METHODS: Incisional hernias were created in 14 Yucatan mini-pigs. Animals were randomized to undergo laparoscopic hernia repair either with spiral tacks alone (Tacks) or with tacks and 4 Prolene transfixion sutures (Sutured) using Composix E/X mesh (Davol Inc.). At 4 weeks, exploratory laparoscopy was performed to assess the repair and score adhesions. The abdominal wall was harvested for tensile strength analysis and histologic evaluation. Continuous variables were compared using a two-tailed nonpaired t-test. Results are presented as mean +/- standard deviation. RESULTS: The mean hernia size was 8.5 +/- 0.5 cm by 5.5 +/- 0.7 cm, with no difference between groups. The operative time was significantly longer ( p = 0.006) for the Sutured group (62.1 +/- 16.8 min) than for the Tacks group (32.3 +/- 7.0 min). The number of tacks per repair was equivalent between groups. At necropsy, the mesh in all cases was well incorporated, reperitonealized, and without evidence of migration. No hernias recurred. However, the Sutured group had a significantly ( p < or = 0.05) higher adhesion score (5.4 +/- 3.3) than the Tacks group (2.0 +/ 2.7). The tensile strength of the repair zone was no different between groups (Sutured 4.8 +/- 1.5 N/cm, Tacks 3.8 +/- 1.4 N/cm). On histologic examination, the ratio of inflammatory cells to fibroblasts was similar between groups (Sutured 0.2 +/- 0.6, Tacks 0.2 +/- 0.3). Only 82% of tacks in each group penetrated the fascia, and the depth of tack penetration was similar between groups (Sutured 3.7 +/- 0.3 mm, Tacks 3.9 +/- 0.4 mm). CONCLUSIONS: In a porcine model, the use of transfixion sutures was associated with longer operative times and more adhesions, without improvement in tensile strength or mesh incorporation. A human clinical trial is needed to determine the optimal method of securing abdominal wall mesh. PMID- 14752651 TI - Port function following laparoscopic adjustable gastric banding. PMID- 14752652 TI - Outcome of laparoscopic redo fundoplication. AB - BACKGROUND: To date, there has been no objective evidence for the effectiveness of laparoscopic redo fundoplication. We therefore reviewed our experience and based our analysis on a number of objective parameters. METHODS: We prospectively followed 28 consecutive patients (five men and 23 women; mean age, 48.64 +/- 2.57 years) who required redo fundoplication. These patients were part of a series of laparoscopic Nissen fundoplications done between 1992 and 2001. The indications were recurrent symptoms of gastroesophageal reflux disease (GERD) (21 patients), acute herniation of the wrap (three patients), and chronic paraesophageal hernia (four patients). A diagnosis of recurrent GERD was based on endoscopy, 24-h pH study, manometry, and symptom score evaluation. A diagnosis of paraesophageal and acute herniation was based on contrast swallow studies and/or gastroscopy. RESULTS: Twenty-six redo fundoplications were completed laparoscopically; two were converted to open. The mean operative time was 55.43 +/- 3.81 min. There were no intraoperative complications. The mean hospital stay was 3.0 +/- 0.35 days. Postoperative complications included postoperative pneumonia in one patient. Two patients from the laparoscopic group required a third operation-one for acute herniation of the redo wrap, which was fixed laparoscopically, and the other for acute recurrent paraesophageal hernia, which was fixed via an open transthoracic approach. The mean follow-up after revision is 25.14 +/- 3.48 months, with a significant decrease in acid reflux from 5.01% +/- 0.99 to 0.48% +/- 0.23 ( p < 0.0001), a significant decrease in symptom score from 28.96 +/- 2.93 to 10.75 +/- 2.61 ( p < 0.0001), and a small but significant increase in lower esophageal sphincter (LES) pressure from 13.71 +/- 1.79 to 16.69 +/- 1.50 ( p = 0.04). CONCLUSIONS: Laparoscopic redo fundoplication is technically feasible and clinically effective over a 2-year objective follow-up. Conversion and complication rates are low. PMID- 14752653 TI - Laparoscopic repair of large paraesophageal hernia is associated with a low incidence of recurrence and reoperation. AB - BACKGROUND: Laparoscopic repair of paraesophageal hernia (LRPEH) is a feasible and effective technique. There have been some recent concerns regarding possible high recurrence rates following laparoscopic repair. METHODS: We reviewed our experience with LRPEH from 5/1996 to 8/2002. Large paraesophageal hernia (PEH) was defined by the presence of more than one-third of the stomach in the thoracic cavity. Principles of repair included reduction of the hernia, excision of the sac, approximation of the crura, and fundoplication. Pre- and postoperative symptoms were evaluated utilizing visual analogue scores (VAS) on a scale ranging from 0 to 10. Patients were followed with VAS and barium esophagram studies. Statistical analysis was performed using two-tailed Student's t-test. RESULTS: A total of 166 patients with a mean age of 68 years underwent LRPEH. PEH were type II ( n = 43), type III ( n = 104), and type IV ( n = 19). Mean operative time was 160 min. Fundoplications were Nissen (127), Toupet (23), Dor (1), and Nissen Collis (1). Fourteen patients underwent a gastropexy. One patient required early reoperation to repair an esophageal leak. Mean hospital stay was 3.9 days. At 24 months postoperatively there was statistically significant improvement in the mean symptom scores: heartburn from 6.8 to 0.5, regurgitation from 5.9 to 0.3, dysphagia from 4.0 to 0.5, chest pain from 3.7 to 0.3. Radiographic surveillance was obtained in 120 patients (72%) at a mean of 15 months postoperatively. Six patients (5%) had radiographic evidence of a recurrent paraesophageal hernia (two required surgery), 24 patients (20%) had a sliding hernia (two required surgery), and four patients (3.3%) had wrap failure (all four required surgery). Reoperation was required in 10 patients (6%); two for symptomatic recurrent PEH (1.2%), four for recurrent reflux symptoms (2.4%), and four for dysphagia (2.4%). Patients with abnormal postoperative barium esophagram studies who did not require reoperation have remained asymptomatic at a mean follow up of 14 months. CONCLUSION: LPEHR is a safe and effective treatment for PEH. Postoperative radiographic abnormalities, such as a small sliding hernia, are often seen. The clinical importance of these findings is questionable, since only a small percentage of patients require reoperation. True PEH recurrences are uncommon and frequently asymptomatic. PMID- 14752655 TI - Periarterial application of papaverine during laparoscopic donor nephrectomy improves early graft function after kidney transplantation in pigs. AB - BACKGROUND: Laparoscopic donor nephrectomy (LDN) increases incentives to donation by subjects who might refuse an open operation. However, the incidence of delayed graft function is higher after LDN than after open operation. This may be caused by the reduction of renal perfusion as a result of the raised intraabdominal pressure and mechanically induced renal angiospasm during the operation. We conducted experiments to find out whether the application of papaverine around the renal artery during LDN could improve early graft function after transplantation. METHODS: Renal function was studied in 10 male pigs (weight approximately 25 kg). The left kidney was harvested laparoscopically (intraabdominal pressure 8 mmHg). Five animals were randomly selected to have perivascular application of 50 mg papaverine (treatment group) before preparation of the vessels. In controls no papaverine was used. After LDN and open right nephrectomy the left kidney was autotransplanted. The main outcome measures were volume of urine produced and creatinine clearance during the first 20 h after the transplant. RESULTS: The groups were comparable in respect of body weight, hemodynamic values, amount of infusions, warm and cold ischemia time, and duration of anastomosis. Urine output and creatinine clearance were significantly higher in pigs treated with papaverine than in controls. CONCLUSIONS: Papaverine substantially improved early graft function in pigs when applied around the renal artery during LDN. Whether this is applicable to procurement of human kidneys remains to be evaluated. PMID- 14752656 TI - Dual endoscopic-assisted endoluminal colostomy reversal: a feasibility study. AB - BACKGROUND: Emergent colostomies are associated with increased morbidity related to second closure operations. The purpose of this canine pilot study was to create a minimally invasive procedure that would reduce the time interval and morbidity involved with colostomy reversals after left colon end colostomies. METHODS: Six mongrel dogs underwent modified laparoscopic Hartmann's procedures in which the stapled end of the rectal stump was approximated to the left colon proximal to the stoma. After 1 week, they underwent an endoluminal colostomy reversal with a computer-mediated, circular stapling device and varying anvil insertion methods. Variables recorded included anvil insertion technique and feasibility, OR time, complications, and number of days to first meal and bowel movement. A contrast enema performed 1 week post colostomy reversal ruled out anastomosis leaks and stenosis. The dogs were euthanized and subjected to necropsy. RESULTS: Of four anvil insertion techniques tested, the most feasible employed a large-bore needle to perforate through the stapled end of the Hartmann pouch into the lumen of the left colon. Simultaneous endoluminal views of the rectal stump with a sigmoidoscope and the left colon lumen with an endoscope permitted a controlled and safe needle puncture. Through the needle, a guide wire was inserted to withdraw the anvil via the colostomy into place. A transanally inserted stapler was then married to the anvil under fluoroscopic guidance, thus completing the anastomosis. The colostomy was then taken down and transected at the level of the colocolostomy. Average operating time was 126 min (range 90 180), diet was tolerated within 1.5 days, and average number of days to first bowel movement was 2.5. The absence of stenosis, leaks, and inadvertent visceral injuries confirmed feasibility. CONCLUSIONS: In this canine model, a dual endoscopic-assisted colostomy reversal with a computer-mediated, circular stapling device is feasible. Using this technique, colostomy reversals can possibly be performed 1 week post-colostomy without entering the peritoneal cavity, thus reducing the number of invasive operations and subsequent morbidity required to manage emergent colon perforations. PMID- 14752657 TI - Staple line coverage with absorbable mesh after thoracoscopic bullectomy for spontaneous pneumothorax. AB - BACKGROUND: Thoracoscopic simple bullectomy for primary spontaneous pneumothorax (PSP) has a relatively high postoperative recurrence rate and sometimes results in postoperative air leakage. One of the reasons for postoperative recurrence is the regrowth of bullae around the staple line. Therefore, reinforcement of the visceral pleura around the staple line is a reasonable way to prevent postoperative air leaks and recurrence. This study was done to determine the efficacy in preventing postoperative air leak and recurrent pneumothorax of widely covering the staple line with absorbable mesh after thoracoscopic bullectomy. METHODS: Wide coverage of the staple line with absorbable mesh was performed on 114 patients with PSP. These patients were retrospectively compared with 126 patients who underwent thoracoscopic simple bullectomy alone. RESULTS: The postoperative duration of chest drainage in the coverage group (mean, 1 day; range, 0-5) was significantly shorter than that in the simple bullectomy group (mean, 3 days; range 0-20). A prolonged air leak (>7 days) occurred in six patients in the simple bullectomy group, but there were no such leaks in the coverage group. Recurrent pneumothorax occurred in three patients (2.6%) in the coverage group and 12 patients (9.5%) in the simple bullectomy group. CONCLUSION: Wide coverage of the staple line with absorbable mesh is effective in preventing postoperative air leak and in decreasing the recurrence rates of PSP. PMID- 14752658 TI - Impact of miniprobe ultrasonography on planning of minimally invasive surgery for gastric and colonic tumors. AB - BACKGROUND: The use of minimally invasive procedures for the management of gastrointestinal cancer is increasing. The aim of this study was to investigate the role of high-frequency miniprobe endoscopic ultrasound (EUS) for therapeutic decisions making in patients with gastric or colonic tumors. METHODS: A total of 137 patients underwent EUS with a 12.5-MHz miniprobe for preoperative staging of tumors of the stomach ( n = 49) or colon ( n = 88). After resection, the surgical path was reviewed to analyze the role of preoperative staging with miniprobes. RESULTS: Miniprobe EUS enabled accurate assessment of the infiltration depth of gastric and colonic tumors. The overall accuracy rates were 88% and 87%, respectively. The lymph node status was predicted correctly in 82% of the patients (sensitivity, 61%, specificity, 94%). Based on the results of miniprobe EUS, patients with gastric cancer were accurately selected to undergo endoscopic mucosal resection, laparoscopic resection, or open surgery in 100%, 91%, and 86% of the cases, respectively. In patients with colonic tumors, the treatment decision analysis showed that the stratification was correct in 90% of the patients. CONCLUSIONS: Miniprobe EUS is a reliable method for validating treatment decisions for patients undergoing minimally invasive procedures for gastric and colonic tumors. This method is particularly valuable in the management of colon cancer, because endoscopic and laparoscopic resections can be offered to selected patients as an alternative to open surgery. PMID- 14752661 TI - Cloning and characterization of the histidine kinase gene Dic1 from Cochliobolus heterostrophus that confers dicarboximide resistance and osmotic adaptation. AB - A gene for a putative two-component histidine kinase, which is homologous to os-1 from Neurospora crassa, was cloned and sequenced from the plant-pathogenic fungus Cochliobolus heterostrophus. The predicted protein possessed the conserved histidine kinase domain, the response regulator domain, and six tandem repeats of 92-amino-acids at the N-terminal end that are found in histidine kinases from other filamentous fungi. Introduction of the histidine kinase gene complemented the deficiency of the C. heterostrophus dic1 mutant, suggesting that the Dic1 gene product is a histidine kinase. Dic1 mutants are resistant to dicarboximide and phenylpyrrole fungicides, and they are sensitive to osmotic stress. We previously classified dic1 alleles into three types, based on their phenotypes. To explain the phenotypic differences among the dic1 mutant alleles, we cloned and sequenced the mutant dic1 genes and compared their sequences with that of the wild-type strain. Null mutants for Dic1, and mutants with a deletion or point mutation in the N-terminal repeat region, were highly sensitive to osmotic stress and highly resistant to both fungicides. A single amino acid change within the kinase domain or the regulator domain altered the sensitivity to osmotic stress and conferred moderate resistance to the fungicides. These results suggest that this predicted protein, especially its repeat region, has an important function in osmotic adaptation and fungicide resistance. PMID- 14752662 TI - The ancestral role of COE genes may have been in chemoreception: evidence from the development of the sea anemone, Nematostella vectensis (Phylum Cnidaria; Class Anthozoa). AB - An orthologue of the COE family of helix-loop-helix transcription factors was recovered from the anthozoan Nematostella vectensis (Cnidaria). NvCOE has high sequence similarity to vertebrate and invertebrate orthologues in all three major functional domains of the protein. In situ hybridization studies show early expression through the cleavage period but transcripts are down regulated at gastrulation while remaining expressed at high levels only in the apical tuft of cilia at the aboral end of the planula larva. It is likely that one of the ancestral roles of the COE family of genes may have been in the development of chemosensory neurons. PMID- 14752663 TI - A feature-integration account of sequential effects in the Simon task. AB - Recent studies have shown that the effects of irrelevant spatial stimulus response (S-R) correspondence (i.e., the Simon effect) occur only after trials in which the stimulus and response locations corresponded. This has been attributed to the gating of irrelevant information or the suppression of an automatic S-R route after experiencing a noncorresponding trial-a challenge to the widespread assumption of direct, intentionally unmediated links between spatial stimulus and response codes. However, trial sequences in a Simon task are likely to produce effects of stimulus- and response-feature integration that may mimic the sequential dependencies of Simon effects. Four experiments confirmed that Simon effects are eliminated if the preceding trial involved a noncorresponding S-R pair. However, this was true even when the preceding response did not depend on the preceding stimulus or if the preceding trial required no response at all. These findings rule out gating/suppression accounts that attribute sequential dependencies to response selection difficulties. Moreover, they are consistent with a feature-integration approach and demonstrate that accounting for the sequential dependencies of Simon effects does not require the assumption of information gating or response suppression. PMID- 14752664 TI - Spatial interference and response control in sequence learning: the role of explicit knowledge. AB - In several sequence learning studies it has been suggested that response control shifts from the stimuli to some internal representation (i.e., motor program) through the learning process. The main questions addressed in this paper are whether this control shift is related to explicit knowledge and whether the formation of these internal representations depends on the stimulus attributes. In one experiment we compared the learning of a response sequence triggered by either spatial location or location symbol (left-right) by using a serial response task (SRT). Symbols were presented at either a centered or random location. The results showed that in the symbolic conditions the shift of response control correlated with the emergence of explicit knowledge. Only participants with complete explicit knowledge seemed to learn the sequence structure beyond probabilistic information (response time "RT" did not depend on the frequency of the response). Moreover, these participants were able to overcome, when needed, spatial interference (RT was the same for both spatially corresponding and non-corresponding trials). However, when spatial location was relevant, RT was always faster, especially for more frequent responses. These results suggest that the relevant stimulus dimension (location or symbol) seems to engage different sequence learning mechanisms. PMID- 14752665 TI - Early signs of lung fibrosis after in vitro treatment of rat lung slices with CdCl2 and TGF-beta1. AB - Precision-cut rat lung slices have been employed in combination with an extensive immunohistochemistry of paraffin-embedded slices for monitoring of early pathohistological changes after exposure to CdCl(2)/TGF-beta(1). Three days of CdCl(2) exposure in combination with TGF-beta(1) seem to be sufficient to induce lung injury with alterations similar to changes observed in early lung fibrogenesis: (1) extracellular matrix accumulation and myofibroblast transdifferentiation (Sirius red staining, collagen type IV, alpha-smooth muscle actin), (2) type I cell injury with loss of type I cell antigens (T1alpha antigen, aquaporin-5, RAGE), (3) increased apoptosis of pulmonary cells (active caspase-3, vimentin cleavage product V1 of caspase-9), and (4) activation of microvascular endothelial cells (podocalyxin, caveolin-1). Western blot analysis confirmed the increasing amount of alpha-smooth muscle actin, the loss of T1alpha antigen, and the increase in caveolin-1 immunoreactivity. The explant culture using CdCl(2)/TGF-beta(1) provides a suitable tool for the study of other factors involved in pulmonary pathology including transcription factors, cytokines, and other metabolites involved in early stages of fibrogenesis. PMID- 14752666 TI - Protein expression pattern of collagen type XV in mouse cornea. AB - PURPOSE: To examine the alteration of protein expression pattern of collagen type XV in cornea during embryonic development and adult tissue repair. Collagen type XV is a basement membrane collagen of a subfamily of multiplexins (multiple triple helix domains and interruptions). Its COOH-terminal peptide has an anti angiogenic effect and its distribution in avascular tissue of cornea is of interest. METHODS: Eyes of mouse embryos [day (E) 10.5-18.5] and healing adult mouse corneas following either debridement injury or incision were embedded in paraffin. Deparaffinized sections were processed for immunofluorescent staining with anti-collagen XV antibody. RESULTS: At E14.5 embryonic corneal epithelium, as well as fibroblasts in eyelids, began to express this collagen type very faintly, and at E18.5, besides corneal epithelial expression, epidermis, palpebral conjunctiva, and keratocytes started to express collagen type XV. In adult mouse cornea, collagen type XV was observed in basal and suprabasal epithelial cells and stroma, but not in the subepithelial basement membrane. Healing epithelial cells following debridement or incision injury down-regulated its protein expression. CONCLUSIONS: Mouse embryonic corneal epithelium and keratocytes begin to express collagen type XV before birth. Healing murine corneal epithelium down-regulates collagen XV expression. The presence of collagen XV in corneal stroma may play a part in avascularity. PMID- 14752667 TI - Second line therapy with weekly low-dose docetaxel for pretreated non-small-cell lung carcinoma patients: a multicenter Italian phase II study. AB - Docetaxel is one of the most active drugs in second-line therapy for non-small cell-lung-carcinoma (NSCLC). The aim of this multicenter study was to evaluate the safety and efficacy of weekly low-dose docetaxel. Forty-two patients with advanced NSCLC pretreated with cisplatinum-based chemotherapy were enrolled. Docetaxel was administered at a dose of 25 mg/m(2) weekly for 12 consecutive weeks. A total of 386 doses were given with a median number of 10 doses per patient (range: 3-12). Treatment showed low incidence of hematologic toxicity and modest non-hematologic toxicity. An episode of grade 4 thrombocytopenia was reported but no episodes of grade 3 or 4 neutropenia. Most frequent non hematologic toxicities were asthenia and alopecia. Response rate was 10.5% and median survival time (MST) was 12.8 weeks. Weekly treatment with 25 mg/m(2) docetaxel for 12 consecutive weeks appears to be a feasible and active regimen with mild toxicity in heavily pretreated NSCLC patients. PMID- 14752668 TI - The relationship between HRCT and pulmonary function in Behcet's disease. AB - The purpose of this study was to compare the pulmonary functions in female Behcet's patients with or without pathological pulmonary lesions using high resolution computerized tomography (HRCT). Twenty-nine female patients aged 19-54 yrs and 20 healthy females aged 19-50 yrs (control group) were accepted into the study. HRCT images were taken and according to HRCT patients were divided into HRCT (+) and HRCT (-) groups (18 patients in each). Pulmonary function tests (PFT) through a spirometer were performed and FVC, FEV(1), FEF(25-75%), PEF, VC, RV, FRC, DLCO and DLCO/VA were determined. No statistically significant difference was observed when the PFT values were compared between HRCT (+) and HRCT (-) patients for the obtained results and the percentage of the expected values. When the PFT values were compared for both HRCT (+) and HRCT (-) patients with the control group separately there was no statistical difference between the best values. A statistically low DLCO/VA value was observed between the percentage of the expected values. It is concluded that though there is a pulmonary restriction in Behcet's disease, this restriction has no relation to pulmonary functions. It would be useful to perform pulmonary function tests in the patients with Behcet's disease both with or without any pathological findings in HRCT for obtaining information about pulmonary functions. We suggest that even when the results of the pulmonary function tests are normal, considering some pathological changes in HRCT, HRCT investigations may be useful for following up the disease. PMID- 14752669 TI - Surfactant lavage with lidocaine improves pulmonary function in piglets after HCl induced acute lung injury. AB - Acute respiratory distress syndrome (ARDS) is associated with significant morbidity and mortality. The pathophysiology of ARDS includes abnormalities of surfactant function as well as pulmonary inflammation. Immunomodulating drugs, like Lidocaine, have shown some success in decreasing inflammation in ARDS. We attempted to combine surfactant lavage's ability to reverse the surfactant dysfunction, while acting as a vehicle to deliver Lidocaine. Gravity-driven surfactant (Infasurf) lavage (35 ml/kg) was administered alone or mixed with Lidocaine after severe HCl acid injury (0.3 N; 3 cc/kg) in neonatal piglets. Treatment groups included: control (C) ( n = 5), surfactant lavage (SL) (35 ml/kg diluted Infasurf) ( n = 7) and SL mixed with Lidocaine (SL+L) ( n = 7). About 26 27% of the lavage was retained (phospholipid 73-74 mg/kg; Lidocaine 1.8 mg/kg). Oxygenation progressively increased in the SL and SL+L groups over the 4-hour period (at 240 min: C = 99 +/- 14; SL = 154 +/- 39; SL+L = 230 +/- 40 mmHg) ( p < 0.05). PaCO(2) increased in all groups from 43 +/- 0.3 to 55 +/- 0.7 mmHg. Only SL+L showed a reduction in PaCO(2) (at 240 min: C = 54 +/- 4; SL = 53 +/- 7; SL+L = 49 +/- 2 mmHg) ( p < 0.05). Finally, SL and SL + L had superior characteristics during the quasi-static pressure volume (PV) procedure as compared to Control ( p < 0.05). In our HCl ALI model, SL improved oxygenation and quasi-static lung compliance over C. The pulmonary function effects of SL were further enhanced by the addition of Lidocaine to the surfactant suspension. Combining therapeutic agents with surfactant lavage may be an effective strategy in ALI. PMID- 14752670 TI - Cholinergic innervation of BALT (bronchus associated lymphoid tissue) in rat. AB - The presence and distribution of acetylcholinesterase (AChE) and cholineacetyl transferase activities (Chat) were examined in the bronchus-associated lymphoid tissue (BALT) of juvenile, adult and old rats. Histoenzymatic and immunochemical methods were used in association with quantitative analysis of images and statistical analysis of the data. Our results showed that both AChE and Chat activities were primarily confined to the BALT lymphoid cells. Only a low level of activity was observed in the sub-pleural parenchyma of the lung and in the wall of the bronchus. Moreover, both AChE and Chat activities in the BALT are specifically located in the lymphoid cells. Histoenzymatic staining and corresponding values of quantitative analysis of images confirmed morphological and immunochemical results. Finally, the intensity of histoenzymatic staining for AChE and of immunochemical staining for Chat in BALT of rats strongly decreases with age. On the basis of our results we hypothesize that both AChE and Chat activities may play an important role in BALT and both these enzymes undergo specific age-related changes. PMID- 14752671 TI - A probative approach for noninvasive evaluation of airway hyperresponsiveness and remodeling in adult asthmatics. AB - We propose a probative approach for noninvasive evaluation of airway hyperresponsiveness (AHR) and remodeling to investigate their outcome in adult asthmatics treated according to the Global Initiative for Asthma (GINA) guideline. Pulmonary function and AHR to methacholine were measured twice with an interval of 24.3 +/- 3.4 months in 18 adult asthmatics during the ongoing treatments. Mathematical formulas previously used in an animal model were applied in human asthmatics to eliminate the effect of airway wall thickening on respiratory resistance (Rrs), calculating indices for the proportional changes with time in airway wall thickness (PW(1)/PW(0)) and airway smooth muscle shortening (PMS(1)/PMS(0)), respectively. The minimum cumulative dose of methacholine (Dmin), an ordinary index of AHR measured with the oscillometry Asthograph, correlated with the asthma severity. The disease periods significantly correlated with the indices of airflow limitation. While there was no change in PW(1)/PW(0) (1.00 +/- 0.07) during the assessment periods, methacholine-induced airway smooth muscle shortening was attenuated by 46% (PMS(1)/PMS(0)=0.54 +/- 0.16). Less improvement in PMS(1)/PMS(0) was seen with a correlation to the disease periods, but PMS(1)/PMS(0) improved correlating to the relative length of the assessment period with ongoing treatments in the disease period. In conclusion, this probative approach may be useful to investigate the outcome of AHR and remodeling in human asthmatics, and shows that remodeling may get worse with time or may halt and AHR may improve with a stepwise, early intervention and prolonged treatment given according to the GINA guideline. PMID- 14752672 TI - Long-term follow-up of pulmonary function in bronchial asthma patients treated with pranlukast. AB - Clinical studies have shown that pranlukast, a selective cysteinyl leukotriene antagonist, is effective for bronchial asthma. In the present paper, we retrospectively analyzed long-term asthma control by pranlukast treatment in patients treated with inhaled corticosteroids. We analyzed medical records and asthma diaries of 21 patients (9 males, 12 females) (52.1 +/- 3.5 years of age) with bronchial asthma who experienced increase of more than 10 L/min in peak expiratory flow in the first 4 weeks of treatment with pranlukast (450 mg/day) and were subsequently treated with pranlukast for more than 1 year. They all received inhaled corticosteroids (400-1600 microg/day of beclomethasone dipropionate or equivalent). We examined clinical control in terms of time course of self-monitored peak expiratory flow. During the analyzed period, the dose of inhaled corticosteroids was tapered in 4 patients, constant in 15 patients and increased in 2 patients. In 19 patients treated with unchanged or tapered dose of inhaled corticosteroids, improvement in the increase of mean PEF at 4-week treatment was maintained for 1 year. No difference in the add-on effect of pranlukast was observed in patients treated with less than 800 microg and more than or equal to 800 microg of inhaled corticosteroids. Four patients underwent reduction of inhaled corticosteroids in the analyzed period and PEF was well maintained and even increased by pranlukast treatment. In 11 patients in whom data for 3 years were available, the improvement in PEF persisted for 3 years. Although the present investigation is a retrospective analysis, these data may suggest that pranlukast has no tachyphylaxis and its effect continues for more than 1 year. PMID- 14752673 TI - Helix wire osteosynthesis for proximal humeral fractures: unacceptable nonunion rate in two- and three-part fractures. AB - INTRODUCTION: Fractures of the proximal humerus are very common, but controversy still exists about the preferred type of operative treatment in displaced fractures. MATERIALS AND METHODS: We followed 15 patients (12 female and 3 male, average age 70 years) with 15 dislocated type II (9) and III (6) proximal humeral fractures after helix wire osteosynthesis. RESULTS: A nonunion developed in seven cases (47%). Three patients were reoperated (prosthesis: 2, repair: 1). Seven patients were available for follow-up (average: 14 months). Three patients had a Constant score of 80 or more and four patients had a score under 70 points. CONCLUSION: The results of the helix wire osteosynthesis for proximal humeral fractures are poor and we do not recommend its further use. PMID- 14752674 TI - Clinical significance of signet ring cell rectal carcinoma. AB - BACKGROUND AND AIMS: Signet ring cell carcinoma of the rectum (SCCR) is a rare type of rectal carcinoma. This study examined the clinical significance of SCCR. PATIENTS AND METHODS: From our medical records we retrospectively identified 61 SCCR patients and compared their clinical data and outcomes to those of 144 consecutive patients with non-SCCR mucinous rectal adenocarcinomas (NSMR) and 2,414 consecutive patients with nonmucinous rectal adenocarcinomas (NMR). RESULTS: . The incidence of SCCR was 1.39% of rectal cancers. Mean patient age at onset of SCCR (48.1years, range 15-80) was significantly lower than that for NSMR (57.4 years, 9-88) and NMR (62.6 years, 12-94). The proportion of late stage (TNM III+IV) tumors was significantly higher in SCCR (90%) than in NSMR (69%) and NMR (48%). There were more tumors located in the lower rectum in SCCR (46%) than in NSMR (34%) and NMR (29%). SCCR tumors were significantly larger (5.68+/-3.84 cm) than NSMR (4.27+/-1.78 cm) and NMR tumors (3.76+/-1.71 cm). A higher percentage of patients with SCCR (42.6%) received abdominoperineal resection for treatment. In tumors with TNM stage IV the rate of tumor spread via the hematogenous route was significantly lower in SCCR (18.5%) than in NSMR (43.5%) and in NMR (69%). The rate of tumor spread via seeding to the peritoneum was lower in SCCR (22.2%) than in NSMR (43.5%) but higher than in NMR (2.7%). The rate of tumor spread via the lymphatic route was higher in SCCR (44.4%) than in NSMR (26.1%) and significantly higher than in NMR (12.3%). The 1-, 2-, and 5-year overall SCCR survival rates were 73.9%, 36.3%, and 23.3% respectively, which were significantly poorer than those of NSMR and NMR. For the 28 stage III and R0 SCCR tumors the 1-, 2-, and 5-year disease-free survival rates of SCCR were 84.0%, 44.2%, and 30.3%, respectively, which are comparable with general data of stage III rectal cancer in the world. CONCLUSION: Diffuse infiltration of signet ring cells enhances the tendency of mucinous carcinomas of the rectum in more local extension and easier lymphatic spreading but not at peritoneal seeding. Although SCCR had the poorest prognosis, this outcome may be due to the advanced tumor stage rather than histology itself. PMID- 14752675 TI - Anastomotic leakage following low anterior resection: results of a standardized diagnostic and therapeutic approach. AB - BACKGROUND AND AIMS: This study analyzed the results of a standardized approach in anastomotic leakage following low anterior resection for rectal cancer without performance of a protective ileostomy during the primary operation. PATIENTS AND METHODS: The study included all 306 patients with rectal cancer electively undergoing low anterior resection with retroperitonealization of the anastomosis over 9 years. The diagnostic procedure for anastomotic leakage included serum laboratory investigations and abdominal CT together with contrast enema. Minor leakages, i.e., small leakages and pelvic abscess, were treated with rectoscopic lavage and/or CT-guided drainage of the abscess, respectively. Major leakage was defined as broad insufficiency with or without septicemia. Nonseptic patients were treated by ileostomy and rectoscopic treatment. In septic patients a revision of the anastomosis with loop ileostomy was performed. RESULTS: Anastomotic leakage was diagnosed in 30 patients (overall 9.8%; 12 major, 18 minor leakages). Common clinical signs were pelvic pain and fever. No patient developed a peritonitis. The most accurate diagnostic instrument was CT (96.7%). CONCLUSION: Retroperitonealization appears to prevent peritonitis in patients with anastomotic leakage following low anterior resection. A differential treatment leads to good results in terms of mortality and anorectal function. PMID- 14752676 TI - A prospective comparison of two regimes of bowel preparation for pediatric colorectal procedures: normal saline with added potassium vs. polyethylene glycol. AB - We prospectively evaluated the safety and efficacy of total gut irrigation (TGI) using normal saline with added potassium (NS) and polyethylene glycol (PG) in patients undergoing a variety of colorectal procedures including single-stage pull-through for Hirschsprung's disease (HD). Fifty-four patients were randomly assigned into one of the two groups (NS or PG). Pre- and post-TGI weight, abdominal girth, and serum electrolytes were assessed. Patients were also evaluated for vomiting and abdominal discomfort. At surgery, bowel preparation was evaluated, and postoperative complications were recorded. Both NS and PG are safe and effective agents for TGI. No patient in either group had any clinical or statistically significant change in the evaluated parameters. The amount of NS required for TGI was significantly higher than PG, and PG was better tolerated than NS. All patients with HD completed TGI, although one patient with long segment HD had abdominal distension. PMID- 14752677 TI - Local dexamethasone improves the intestinal lesions of gastroschisis in chick embryos. AB - BACKGROUND AND AIMS: Eviscerated bowel in gastroschisis (Gx) undergoes changes that lead to dysfunctions and create management difficulties. This study tests the hypothesis that exposure of the eviscerated bowel of chick embryos with Gx to dexamethasone might have beneficial effects on the parietal lesions. METHODS: Gx was created in chick embryos on incubation day 15 and either dexamethasone (0.047 mg in 0.24 ml) or 0.075% saline were instilled into the amnio-allantoic chamber on day 17. The chicks were recovered near hatching (day 19) and eviscerated and non-eviscerated portions of the intestines were recovered, weighed and processed for HE and synaptophysin staining or for total DNA and protein measurements. Total mural and serosal layer thickness were determined and intramural ganglia were counted. ANOVA was used for comparison among groups with significance level set at p<0.05. RESULTS: Chicks with Gx and Gx + saline controls had reduced body weight and tibial length in comparison with controls. The eviscerated bowel was heavier with marked wall thickening at the expense of all layers but particularly of the serosa. They had decreased total intestinal DNA with normal protein and decreased intramural ganglion density. In contrast, chicks from the Gx + dexamethasone group had normal body weight and tibial length, near-normal intestinal wall thickness with slightly increased serosal width, near-normal intestinal DNA content and normal density of intramural ganglia. CONCLUSION: Local dexamethasone had beneficial effects on the eviscerated bowel of chicks with Gx as judged by decreased wall thickening, normalization of total intestinal DNA and richer neural population. Late gestational exposure to steroids could represent another alternative for preventing intestinal lesions in Gx. PMID- 14752678 TI - Stereotactic biopsy of brain tumours in the paediatric population. AB - OBJECTIVE: Our objective was to establish the role of stereotactic neurosurgical techniques in the management of brain tumours in children. MATERIALS AND METHODS: A retrospective analysis was conducted of all stereotactic procedures performed in a single centre between 1996 and 2001. The success rates of achieving histologic diagnosis and a correlation between radiologic and histologic diagnosis were examined. RESULTS: There were 7 boys and 7 girls with a mean age of 9.1 years (range: 4-15). Under general anaesthetic 15 procedures were performed in 14 patients: 12 diagnostic and 3 therapeutic; 10 with CT and 5 with MRI guidance; 10 lesions were supratentorial and 4 were in the pons. A definitive histologic diagnosis was established in 10 of the 12 cases (diagnostic yield 83%). The pre-operative radiological diagnosis was accurate for tumour type in only 75% of the cases. In 3 patients cyst aspiration was attempted: post operatively the cyst size was decreased in 2 and unchanged in 1. Seizures and acute confusion following biopsy of a thalamic tumour occurred in 1 patient. Post operative scans were performed in 7 patients and in 2 we noted small, clinically insignificant, haemorrhages at the biopsy site. There was no mortality and morbidity was 6.6%. CONCLUSIONS: This small series confirms that stereotactic procedures in children are safe, well tolerated, with a high diagnostic yield, which could be improved with the use of intraoperative histopathological examination. PMID- 14752679 TI - [Legal liability and medico-legal problems of surgeons in Turkey]. AB - Medicine, as a profession, is quite a hard commitment, with the main aim of giving care to patients. Physicians assume professional accountability not only to their patients and relatives but also to legal authorities. In case of insufficiency in exercising this multi-dimensional accountability, they may confront legal and ethical issues. A high index of awareness of responsibility and accountability may help them avoid getting involved in unwilling situations. This article revisits physicians' and, in particular, surgeons' legal liability to medical legislation and jurisprudence in Turkey, with emphasis on medico-legal consequences. PMID- 14752680 TI - Comparison of repair techniques in rat duodonal perforations: simple closure, simple closure and omentoplasty, and fibrin tissue adhesive. AB - BACKGROUND: We compared three repair techniques, namely, simple closure and omentoplasty, simple closure alone, and fibrin tissue adhesive, in the treatment of rat duodenal perforations induced in the postpyloric region. METHODS: Thirty male Sprague-Dawley rats (210-240 g) were included. Under ketamine and ether anesthesia, duodenal perforations of 2 mm were induced in all rats in the postpyloric region. The rats were assigned to three groups equal in number, which underwent repair with the use of simple closure and omentoplasty, simple closure alone, and fibrin glue, respectively. All the animals were sacrificed under deep ether anesthesia on the fourth postoperative day. To assess wound strength, bursting pressure measurements were performed. In addition, hydroxyproline content of the wound site was measured by absorbance spectrophotometry at 560 nm. RESULTS: The mean bursting pressure in the simple closure and omentoplasty group was significantly higher than those of the simple closure (p<0.05) and fibrin glue (p<0.001) groups. No significant difference existed between the simple closure and fibrin glue groups in this respect (p>0.05). Hydroxyproline values of the simple closure and omentoplasty (p<0.05) and simple closure (p<0.001) groups were significantly lower than that of the fibrin glue group. CONCLUSION: Fibrin tissue adhesives may have an application in the treatment of duodenal ulcer perforations, as an adjunct to laparoscopic or open surgical methods. PMID- 14752681 TI - Surgical treatment of ano-rectal gunshot injuries caused by low-velocity bullets. AB - BACKGROUND: The aim of the study was to evaluate the results of treatment for ano rectal gunshot injuries in civilian population, caused by low-velocity bullets. METHODS: Nine patients with ano-rectal gunshot injuries were admitted to the emergency department. All the patients were males, with a mean age of 23 years (range 20 to 37 years) and presented within the first two hours following injury. Complete physical and abdominal examinations were performed and injury severity scores (ISS) were calculated. Injuries were evaluated by rectosigmoidoscopy in the operating room. Seven patients had associated tissue or organ injuries including bladder disruption, pelvic bone fractures, and wide muscular defects. Surgical procedures included a diverting ostomy, irrigation of distal rectum, presacral drainage (6 patients), and retrorectal drainage through the abdomen (3 patients). Antibiotic prophylaxis was routinely administered. Control examinations were made at the end of the second month. RESULTS: Seven patients had rectal blood discharge on admission. The mean ISS score was 7.3+/-3.7, with only one patient having an ISS of 15. Early postoperative complications were urinary infection in three patients, wound dehiscence in five patients, and osteomyelitis in one patient. Anal continence was not adversely influenced after surgery. CONCLUSION: Our results suggest that a diverting ostomy, distal rectal irrigation, and presacral drainage yields favorable outcome in patients with ano rectal gunshot injuries. PMID- 14752682 TI - [Surgical treatment of iatrogenic cardiac traumas induced by heart catheterization]. AB - BACKGROUND: A retrospective evaluation was made on iatrogenic cardiac traumas requiring surgical treatment, that were induced by cardiac catheterizations and interventions performed within a 17-year period. METHODS: A total of 64,911 patients underwent cardiac catheterizations and interventions from 1985 to 2002. Complications of iatrogenic cardiac traumas induced by these interventions were examined together with the surgical treatment performed within 24 hours after catheterization. RESULTS: Iatrogenic cardiac trauma requiring prompt surgical intervention was documented in 20 patients (6 females, 14 males; mean age 51 years; range 31 to 69 years). These were due to coronary angiography/balloon angioplasty-stenting in 14 (70%), percutaneous mitral balloon valvuloplasty in four (20%), and to heart catheterization in two patients (10%). Acute cardiac tamponade was detected in 10 patients (50%) resulting from perforations to the cardiac chambers in six, coronary arteries in two, and major vessels in two patients. Surgical interventions included coronary artery by-pass in 14, mitral valve surgery in four, and repair of major vessels in two patients. Perioperative mortality occurred in two patients. Six patients developed complications contributing to morbidity, including perioperative myocardial infarction (3 patients), infection (2 patients), and prolonged intubation (1 patient). CONCLUSION: In case of major cardiac complications induced during cardiac catheterizations, early diagnosis and prompt surgical intervention are of vital importance regardless of considerably high risks. PMID- 14752683 TI - [Traumatic vascular complications due to intraaortic balloon pump support]. AB - BACKGROUND: The use of an intraaortic balloon pump (IABP) catheter was retrospectively evaluated in terms of risk factors, insertion techniques, and complications in patients with low cardiac output. METHODS: A total of 1036 patients (804 males, 232 females; mean age 53.4 years; range 16 to 75 years) received IABP support from 1985 to March 2002. Of these, 789 patients (76.1%) underwent open heart surgery, 247 patients (23.8%) developed low cardiac output during medical treatment. Insertion of IABP was performed via the femoral artery either percutaneously by the Seldinger technique in 897 patients (86.6%), or by direct surgical exposure in 88 patients (8.5%). Open surgical IABP insertion was performed through an 8 mm Dacron graft placed with an end-to-side anastomosis to the common femoral artery (88 patients) or to the ascending aorta (23 patients). RESULTS: The overall mortality rate was 35.1% (364 patients). Vascular complications were associated with IABP in 104 patients (10%), of which 57 patients (5.5%) required surgical treatment. Major complications were aortic arch dissection in two patients and paraplegia in two patients. Vascular complications tended to increase with female gender, older age, diabetes, and peripheral vascular disease. The mean duration of IABP support in the presence of vascular complications was 7.8 days (range 5 hours to 77 days). CONCLUSION: Application of unsheathed IABP and proper evaluation of peripheral circulation seem to decrease the incidence of vascular complications. PMID- 14752685 TI - Seasonality in the incidence of abdominal aortic aneurysm ruptures: a review of eight years. AB - BACKGROUND: Ruptured abdominal aortic aneurysm (RAAA) is a highly mortal entity. In recent years, the role of seasonality has been proposed in the incidence of RAAA. In this study, we sought possible correlations between monthly atmospheric pressures and the number of patients admitted with RAAA. METHODS: Twenty-four patients who were admitted to our Trauma and Emergency Surgery Department with a diagnosis of RAAA from January 1995 to May 2003 were retrospectively evaluated. Data were collected from patient records, admission charts, hospital death certificate registry, and operating-room records. Only patients whose diagnosis of RAAA was confirmed during surgery were included. Atmospheric pressure records of Istanbul for individual months covering the study period were obtained from the Meteorology Office. The incidences of RAAA and the mean monthly atmospheric pressures were compared. RESULTS: Most of the patients were admitted in winter months. Admissions culminated in January with six patients, at which time the mean atmospheric pressure was 765.5 mmHg. There were no admissions in May, during which the mean atmospheric pressure was 760.8 mmHg. The distribution of monthly admissions for RAAA was not statistically significant (p>0.05). The mean atmospheric pressures tended to run a higher course before the months in which increased admissions were seen. CONCLUSION: Our findings do not corroborate the presence of a relationship between the incidence of RAAA and the atmospheric pressure. PMID- 14752684 TI - [The efficacy of local T-pa infusion in the treatment of axillary vein thrombosis]. AB - BACKGROUND: We evaluated the results of local tissue plasminogen activator (t-PA) infusion in the treatment of axillary vein thrombosis. METHODS: Fourteen male patients (mean age 23+/-2 years) who presented with pain and swelling in the arm were diagnosed as having axillary vein thrombosis. Besides physical examination, diagnoses were made with the use of venous Doppler ultrasound, duplex scanning or venography. Complaints were localized in the right upper arm in 10 patients, and in the left upper arm in four patients. Time to presentation from the onset of complaints ranged 1-3 days, 3-7 days, 7-10 days, and 10-14 days in six, three, two, and three patients, respectively. Infusion of t-PA (0.25 mg/kg) was performed for one hour via a venous catheter inserted to the distal part of the brachial vein. Venous patency was evaluated using Doppler ultrasound and venography. RESULTS: Axillary vein was shown to be patent by venous Doppler ultrasound in eleven patients within a mean of 4.2 hours. Three patients who exhibited no improvement received another t-PA (0.05 mg/kg/h) infusion six hours after the initial treatment. Venous Doppler ultrasound showed almost complete patency of the lumen after six, seven, and 11 hours of the subsequent infusion, respectively. Following therapy, all the patients were administered prophylactic anti-aggregating treatment. On control examinations after three months, venous Doppler ultrasound and venography showed that the lumens were almost completely patent in 12 patients and two patients, respectively. No recurrences were detected. CONCLUSION: Fibrinolytic agents yield high efficacy in the treatment of axillary vein thrombosis. PMID- 14752686 TI - [Thoracic trauma: an analysis of 521 patients]. AB - BACKGROUND: We evaluated thoracic trauma cases with regard to etiologic causes, treatment methods, outcome, and factors affecting the results in the light of our experience and relevant literature data. METHODS: A retrospective evaluation was made of 521 patients (399 males, 122 females; mean age 42 years; range 4 to 93 years) who were treated for thoracic trauma from September 1997 to August 2002. RESULTS: Isolated thoracic trauma and multisystem trauma were found in 348 (67%) and 173 (33%) patients, respectively. Blunt injuries accounted for 87%, the most common cause being traffic accidents (62%), and penetrating injuries accounted for 13%. The most frequent thoracic pathologies included multiple (56%) and single (24%) rib fractures, and flail chest (8.4%). Extrathoracic injuries were seen in 33.2%, the extremities (41%) and the skull (40%) being the most commonly involved. Treatment consisted of symptomatic treatment in 159 patients (30.5%), tube thoracostomy in 337 patients (64.6%), and thoracotomy in 15 patients (2.9%). In isolated thoracic trauma and multisystem trauma, morbidity rates were 3.4% and 22%, and mortality rates were 2.5% and 16.2%, respectively. The overall mortality was 7.1% (37 patients). The mean length of hospital stay was 7.1 days (range 1 to 64 days). CONCLUSION: A great majority of thoracic trauma patients can be treated with conservative methods or tube thoracostomy. Accompanying multisystem traumas are associated with dramatic increases in morbidity and mortality rates. PMID- 14752687 TI - [A retrospective analysis of 204 mandibular fractures]. AB - BACKGROUND: We retrospectively reviewed patients who were treated and followed-up for mandibular fractures within a 10-year period. METHODS: A total of 204 patients (158 males, 46 females; mean age 22.4 years; range 5 to 72 years) were retrospectively evaluated with respect to age groups, sex, etiology, associated injuries, localization and type of the fractures, treatment methods, and early and late complications. The follow-up period ranged from four months to 10 years. RESULTS: The most common cause of injury was traffic accidents (44.1%), followed by falling (31.8%), and violence (17.1%). The highest incidence occurred at ages 21 to 30 years. Of 283 fractures detected, the most common fracture sites were the parasymphysis (83 fractures, 29.3%) and the angulus (52 fractures, 18.4%). Forty-eight patients (24%) had associated injuries. The type of the fractures was simple in 80 patients (39.2%), and complex in 62 patients (30.4%). Treatment included open reduction with titanium mini-plates and screws in 130 patients, and intermaxillary fixation in the remaining patients. The fractures recovered without any complications in 167 patients (81.8%). No occlusion-related complications occurred in the late follow-ups. Complications were encountered in 37 patients (18.1%), being in the early (malocclusion in 5.9%, infections in 2.5%, inferior alveolar nerve injuries in 2.5%) or late (plate-screw exposition in 4.4%, ankylosis of the temporomandibular joint in 2%, and orocutaneous fistula in 1%) postoperative periods. CONCLUSION: Rigid fixation should be the first choice of treatment in mandibular fractures. PMID- 14752688 TI - [The results of nerve repair in combined nerve-tendon injuries of the forearm]. AB - BACKGROUND: We evaluated the clinical and functional results of nerve repair in patients with combined tendon-nerve injuries of the forearm. METHODS: The study included 68 patients (58 males, 10 females; mean age 33.5 years; range 5 to 48 years) with combined tendon-nerve injuries of the forearm. A total of 96 nerves were repaired. Patients with nerve defects were excluded. Both median and ulnar nerves were injured in 17 patients; median and ulnar nerve injuries were detected in 17 patients and 34 patients, respectively. The mean time to operation was four hours (range 20 min to 24 h). Primary nerve repair was performed in 60 patients, and secondary repair was performed in eight patients. The interfascicular technique was employed in 18 patients, and epiperineural suture in 50 patients. Rehabilitation included early motion using the Washington regimen. A modified MRC (Medical Research Council) motor and sensory classification system was used for postoperative evaluation. The mean follow-up was at least two years. RESULTS: The modified MRC results were as follows: of 17 patients with median nerve injuries, 10 had excellent, seven had good results. Of 34 patients with ulnar nerve injuries, the results were excellent in nine, very good in 10, good in 10, and fair in five patients. Of those with median and ulnar nerve injuries, four, seven, and six patients had excellent, very good, and good results, respectively. Overall, 61 patients (89.7%) had satisfactory results. A significant correlation was found between age and the MRC results (p=0.016). CONCLUSION: Primary nerve repair followed by early motion results in substantial rates of excellent and satisfactory results in patients with combined nerve-tendon injuries. PMID- 14752689 TI - [Traumatic aortic rupture: a case report]. AB - Aortic injuries can easily be missed in polytraumatized patients due to either associated injuries or vagueness of physical examination findings. Especially in young adults without associated injuries, relatively low atherosclerotic changes may limit the dissection of traumatic descending aortic injuries. We present a 34 year-old male patient who only had a complaint of back pain following a traffic accident. Upon detection of no abnormalities on plain X-ray films, he was discharged home with analgesics. One week later, he presented with dypsnea and dysphagia. Aortography showed rupture of the aorta and a pseudoaneurysm near the isthmus. Following surgical excision of the pseudoaneurysm and aortic repair with a Dacron graft interposition he was discharged with complete relief of symptoms. This case emphasizes the need for a high index of clinical suspicion in timely diagnosis and prompt treatment of traumatic aortic ruptures. PMID- 14752690 TI - A case of blunt abdominal trauma and posttraumatic acute appendicitis. AB - Acute appendicitis and blunt abdominal trauma are common surgical emergencies. Whether there is a causative relationship between these two entities has long been a subject of debate. A twenty-one-year-old male Japanese tourist presented with vague abdominal pain and dysuria that began after he had been beaten and robbed. No signs of trauma were detected on physical examination; however, there were diffuse abdominal sensitivity with maximal tenderness in the hypogastrium and rebound tenderness in the right lower quadrant. Upon no improvement with medications within 24 hours, laparotomy was performed, which revealed an inflamed appendix, a few enlarged mesenteric lymph nodes, and free peritoneal fluid that was found to be sterile. Following appendectomy, the diagnosis was confirmed by pathologic examination and the enlargement of the lymph node was attributed to non-specific reactive hyperplasia. The patient had an uneventful postoperative course, with relief of pain and fever. PMID- 14752691 TI - A case of giant appendiceal mucocele. AB - Appendiceal mucocele is a rare clinical condition that causes distension of the appendix lumen with mucus. A seventy-three-year-old female patient presented with complaints of abdominal pain, nausea, and vomiting. Abdominal examination revealed mild tenderness, right lower quadrant pain upon palpation, rebound tenderness and muscular rigidity, and a palpable mass. Abdominal ultrasonography and computed tomography scans demonstrated a cystic lesion in the right iliac fossa, adherent to the cecum, suggesting an abdominal abscess. An emergency operation was performed, during which a diagnosis of a mucocele of the appendix was made. Surgical treatment included appendicectomy, partial resection of the ileum, and resection of the cecum. Histopathologic examination confirmed the operative diagnosis. The role of imaging and clinical approach is emphasized in the treatment of an appendiceal mucocele, especially in emergency settings. PMID- 14752692 TI - Secondary intestinal obstruction due to low-grade mucinous cystadenocarcinoma of the appendix. AB - Appendiceal mucoceles are a group of lesions in which the appendiceal lumen becomes distended with mucus. They may be associated with an unusual form of gelatinous ascites termed pseudomyxoma peritonei. We report herein a 77-year-old male patient with appendiceal mucinous cystadenocarcinoma and pseudomyxoma peritonei, whose initial symptoms were abdominal pain and distension and intestinal obstruction. He had a history of explorative laparotomy for an abdominal mass and ascites 14 months ago at another center. He was suffering from abdominal pain and intestinal obstruction on admission to the emergency service. We performed right colectomy and total resection of the mass which originated from the appendix. Pathologic examination of the specimen showed low-grade mucinous cystadenocarcinoma and pseudomyxoma peritonei. The postoperative period was uneventful. PMID- 14752693 TI - [Management in ophthalmology: a reality of our times]. PMID- 14752694 TI - [Nucleotides: new therapeutic approach for dry eye treatment]. PMID- 14752695 TI - [Influence of filtering surgery on automated perimetry]. PMID- 14752696 TI - [Residual myopia after penetrating keratoplasty for keratoconus]. AB - PURPOSE: To study the results of penetrating keratoplasty for keratoconus analyzing the differences in ultimate spherical equivalent according to graft size and the trephine used in the recipient. MATERIAL AND METHODS: Retrospective study of 38 patients that were divided into three groups. Group A is composed of patients with a 0.50 mm donor-recipient difference, 0.25 mm in group B and same sized in group C. In each group visual acuity, spheric equivalent, keratometry and other parameters were assessed. Statistical analysis was performed by comparing the means of the parameters evaluated. RESULTS: We found a better final visual acuity in group C when compared with group A, and a lower spherical equivalent when we compared groups A with C and A with B. The results were better in those groups where the donor-recipient difference was smaller. CONCLUSIONS: We believe that it is convenient to minimize the size difference between donor and recipient or to use same-sized trephines to diminish the final spheric equivalent. PMID- 14752697 TI - [Conjunctival bacteria of patients undergoing cataract surgery: changes in the last 50 years]. AB - PURPOSE: To describe the prevalence of preoperative conjunctival bacteria of patients living in Madrid, who underwent cataract surgery in our hospital, without using exclusion criteria to select the patients. METHOD: The preoperative conjunctival cultures of 4432 consecutive patients, in a three year period, were studied retrospectively. Samples of preoperative conjunctival cultures were selected from publications of the past 50 years. Our conjunctival bacteria frequencies were compared with those of other samples (chi-square test), in order to show that the bacterial isolation method is a determinant factor of the bacterial spectrum. RESULTS: 93.5% bacteria were Gram positive and 6.5% Gram negative. Each bacteria group was present in the patients as follows: 56.9% Staphylococcus coagulase negative, 30.5% Corynebacterium sp, 6.5% Staphylococcus Aureus, 2.7% Streptococcus Pneumoniae, 5.6% other Streptococcus sp, 2.8% Haemophilus sp, 3.5% other Gram negative rods, 1.2% diplococci Gram negative and 0.5% other bacteria. Pathogen bacteria were isolated in 19.1% of the patients and 21.9% of the cultures were negative. The samples shown in publications previous to 1980 (table II), present bacteria frequencies statistically different from our study. CONCLUSIONS: The conjunctival bacteria prevalence in preoperative cultures of patients awaiting cataract surgery had never been described in Madrid. By analysing other authors' results, it can be seen that bacteria prevalence is affected by the culture media used. It also seems that the size of the sample and the health status of the patients could affect this prevalence (Arch Soc Esp Oftalmol 2004; 79: 13-20). PMID- 14752698 TI - [Correlations between retinal thickness analyzer (RTA) and confocal scanning laser tomography (HRT) in optic disc analysis]. AB - OBJECTIVES: To determine if the measures obtained in an optic disc study using the retinal thickness analyzer (RTA) can be adequately correlated with values provided by the confocal scanning laser Heidelberg Retina Tomograph (HRT). METHODS: We analyzed 20 eyes of 20 normal subjects. The contours of the optic disc were drawn by a single operator using both instruments. The data compared were the values of 10 variables denoted in the same way by each instrument and these provided information on disc and cup morphology. RESULTS: Of the 10 variables, only the following three showed clear correlation between the two instruments: maximum cup depth (r=0.926, p=0.001), mean cup depth (r=0.857, p=0.001) and cup shape measure (CSM, r=0.672, p=0.002). The correlation shown by the remaining variables ranged from 0.001 (cup area) and 0.488 (cup volume). CONCLUSIONS: The variables showing best correlation between instruments were those providing information on cup morphology. Operator subjectivity when drawing the disc contour using each device could account for the lower incidence of correlation of the other variables examined. PMID- 14752699 TI - [Monolayered amniotic membrane transplantation as a palliative treatment for bullous keratopathy]. AB - PURPOSE: To determine the efficacy of monolayered amniotic membrane transplantation for short and medium-term symptomatic improvement in those cases of bullous keratopathy where a penetrating keratoplasty is not indicated. METHODS: Amniotic membrane transplantation was performed in five patients with symptomatic bullous keratopathy in which penetrating keratoplasty was not indicated. Data about bandage contact lens need before and after surgery, existence of ocular pain, occurrence and extension of bullae, epithelial defects, and persistence of amniotic membrane, were obtained at days 4 and 14, and months 1, 2, 4 and 6 after surgery. RESULTS: No pain, bullae or epithelial defects were present at day 14 in any patient. However, progressive dissolvement of the amniotic membrane was accompanied with more bullae and pain. At 6th month, 4 of 5 patients required bandage contact lens. CONCLUSION: Amniotic membrane transplantation alleviates short-term symptomatic bullous keratopathy and preserves corneal anatomy. Six months after surgery, anatomical and symptomatic improvement does not avoid the need of bandage contact lens. PMID- 14752700 TI - [Herpes simplex virus type 1 acute retinal necrosis two years after presumably herpetic meningoencephalitis]. AB - CASE REPORT: We report the case of a 53-year-old woman with uveitis in her right eye. She suffered from meningoencephalitis two years before. In the ophthalmic examination she showed no light perception, mild anterior uveitis and severe vitritis, which prevented from visualizing the retina. We suspected herpetic acute retinal necrosis (ARN), so therapy with intravenous acyclovir was started and a diagnostic vitrectomy was performed. Peripheral retinal necrosis and pallor of the optic disc were observed. PCR of the vitreous was positive for herpes simplex virus type I. DISCUSSION: This is probably a case of brain-to-eye virus transmission. According to this, the ARN would support the etiologic suspicion of the previous encephalitis. PMID- 14752702 TI - [A curious treatment for amaurosis (XVIII Century)]. PMID- 14752701 TI - [Bilateral macular cysts secondary to electric current strike. A case report]. AB - OBJECTIVE: To report the clinical and tomographic findings in a case of maculopathy secondary to high-tension electric current strike. CASE REPORT: A 27 year-old male refers a reduction in his visual acuity after electric strike. Visual acuity was 0.05 in both eyes. Circular reddish macular lesions measuring 400 micro m in RE and 200 micro m in LE were evident. Optical coherence tomography (OCT) showed intraretinal foveal cysts. At the third month follow-up visit, visual acuity had improved to 0.4 in RE and 0.5 in LE. Clinical and tomographic evaluation showed a reduction of the foveal cysts. DISCUSSION: Electricity affects retinal pigment epithelium and retina by thermal denaturation. OCT provides a conclusive diagnosis. PMID- 14752703 TI - [With the authorities and the weather permission]. PMID- 14752704 TI - Increasing the power and efficiency of disease-marker case-control association studies through use of allele-sharing information. AB - Case-control disease-marker association studies are often used in the search for variants that predispose to complex diseases. One approach to increasing the power of these studies is to enrich the case sample for individuals likely to be affected because of genetic factors. In this article, we compare three case selection strategies that use allele-sharing information with the standard strategy that selects a single individual from each family at random. In affected sibship samples, we show that, by carefully selecting sibships and/or individuals on the basis of allele sharing, we can increase the frequency of disease associated alleles in the case sample. When these cases are compared with unrelated controls, the difference in the frequency of the disease-associated allele is therefore also increased. We find that, by choosing the affected sib who shows the most evidence for pairwise allele sharing with the other affected sibs in families, the test statistic is increased by >20%, on average, for additive models with modest genotype relative risks. In addition, we find that the per-genotype information associated with the allele sharing-based strategies is increased compared with that associated with random selection of a sib for genotyping. Even though we select sibs on the basis of a nonparametric statistic, the additional gain for selection based on the unknown underlying mode of inheritance is minimal. We show that these properties hold even when the power to detect linkage to a region in the entire sample is negligible. This approach can be extended to more-general pedigree structures and quantitative traits. PMID- 14752705 TI - [Lipid peroxidation in adult epileptic patients treated with valproic acid]. AB - INTRODUCTION: Free radicals play an important role as regulatory mediators in cellular signalling processes; however, when overproduced or when antioxidant defence systems are weakened, they are cause of cellular damage. Excessive amount of free radical production has been related with a variety conditions, like aging, different kind of diseases, and xenobiotics biotransformation; this last process includes the metabolism of lipid soluble drugs. An increase in oxidative stress has been described in series of treated epileptic patients. OBJECTIVE: To evaluate the susceptibility to plasma lipid peroxidation in samples from epileptic patients treated with valproic acid monotherapy, studying if the formation of lipid peroxides was related with plasma drug concentration, patients' sex or the kind of epilepsy suffered. PATIENTS AND METHODS: Peroxidated lipids (LPO) were measured by spectrofluorometry before and after induction of an oxidative Fenton reaction in 76 epileptic patients and 4 healthy subjects. RESULTS: After induction of the Fenton reaction, but not in basal conditions, lipid peroxidation showed a lineal relationship with valproate plasma levels. Oxidized LPO values were also significantly higher in samples from patients with partial epilepsies than in those with generalized epilepsies. Likewise, a significant gender effect was observed, being values from epileptic women noticeably higher than those of epileptic men. CONCLUSIONS: Plasma from epileptic patients receiving valproic acid evidences an increased vulnerability to lipid peroxidation which seems to be related with drug amount in the body, subject's sex, and epilepsy type. PMID- 14752706 TI - [Microcephalus as the reason for visiting a regional referral neuropaediatric service]. AB - INTRODUCTION: The first contact between the patient and clinician takes place when the former visits because of some health problem. PATIENTS AND METHODS: We carried out a review of the clinical records of children who had visited the Neuropaediatric Service because of, among other reasons, an isolated or associated microcephalus over a period of 12 years and 9 months. Factors that were considered included whether or not there was a cephalic perimeter below p3 and evidence of encephalopathy, as well as its prenatal, perinatal or postnatal origin, functional diagnoses and the aetiological diagnosis. RESULTS: In 58 cases (0.92%) out of a total number of 6257 children the visit was due to microcephalus. The mean age at the last visit was 3.9 years. In five children (8.6%) the cephalic perimeter was not below p3. No encephalopathy was found in 20 patients (34.4%) and 38 (65.5%) were seen to have encephalopathy, 37 with a prenatal origin: nine genetic, three disruptive and 22 unspecified. Functional diagnoses were as follows: mental retardation in 29 patients, infantile cerebral palsy in 18, autistic spectrum in four and epilepsy in four. Neuroimaging studies aided diagnosis in 13 cases, i.e. 43.3% of those carried out. CONCLUSIONS: Visits to the doctor because of microcephalus, as well as in normal children, include the whole range of prenatal encephalopathies and are associated, ordered according to the frequency of occurrence, with mental retardation and with infantile cerebral palsy. Individual evaluation and clinical progression allow the orientation of each case. Neuroimaging is the most useful complementary examination for diagnostic purposes. PMID- 14752707 TI - [Anoxic encephalopathy. Clinical description, prognosis and neuroimaging]. AB - OBJECTIVES: To describe the clinical profile (neuropsychological, psychopathological, functional and neuroimaging), as well as the evolution of patients with anoxic encephalopathy. PATIENTS AND METHODS: Nine patients with anoxic encephalopathy attending our Service were included in the study. All patients were assessed with a broad range of neuropsychological tests, checklists of psychopathological symptoms, and several functional scales. A CT/MRI or a positron emission tomography (PET) were performed in five patients. Five patients were admitted to a multidisciplinary rehabilitation program. RESULTS: All patients showed problems in orientation, executive functions, verbal learning an immediate and long-term verbal memory, in association with diffuse cognitive changes in other functions. Psychopathologically, all patients showed apathy indifference, and eight subjects showed anosognosia. All subjects have an important dependence in daily activities. CT/MRI were normal or showed subcortical changes whereas the PET showed a predominantly cortical hypometabolism with specific patterns. There were no significant improvements after rehabilitation in treated patients. CONCLUSION: In the absence of a unique clinical profile, our patients with anoxic encephalopathy showed similarities in their symptoms (diffuse cognitive deficits with predominance of amnesic and executive impairments; presence of apathy and anosognosia; complete functional dependence; and poor response to the rehabilitation). Functional neuroimaging could be a useful tool for a better understanding of these encephalopathies. PMID- 14752708 TI - [The relation between the HLA-DRB1*1501 allele and the severity of multiple sclerosis in a sample of the Spanish population from the Balearic Islands: the influence exerted by sex]. AB - INTRODUCTION: There is some controversy about the possible relation between HLA DR2 (DRB1*1501-DRB5*0101-DQA1*0102-DQB1*0602) and the severity of multiple sclerosis (MS), which could be due, at least in part, to methodological differences among the different studies dealing with this subject and/or to a lack of phenotypic homogeneity within the series of patients. AIMS: This study aims to contribute information about the possible relation between DR2 (more specifically the HLA-DRB1*1501 allele) and the severity of MS. PATIENTS AND METHODS: We conducted a study of 43 individuals with clinically defined MS, whose degree of severity was determined using Kurtzke's EDSS, and 107 controls from a similar ethnic origin. DETERMINATIONS: HLA typing by PCR-SSO and PCR-SSP, analysis of oligoclonal IgG bands in CSF by isoelectric focusing, and quantification of the intrathecal synthesis of IgG (IgG index, IgG ratio, Reiber's formula and Tourtellotte's formula). STATISTICS: Chi2 test, Mann-Whitney test and Kendall correlation coefficient. RESULTS: DRB1*1501 is associated with the presence of MS only in females and with lower severity of the disease only in males. These associations do not appear to depend on any kind of effect exerted by DRB1*1501 on the intrathecal synthesis of IgG that differs between the two sexes. CONCLUSIONS: The absence of a relation between DRB1*1501 and the severity of MS reported in many studies could be due to not stratifying the patients according to sex. Our findings emphasise how important it is in genetic studies of complex traits to reduce the phenotypic heterogeneity of patients as much as possible. PMID- 14752709 TI - [Hemiparetic cerebral palsy and startle epilepsy]. AB - OBJECTIVES: We analyzed electroclinical features and evolution in nine patients with hemiparetic cerebral palsy associated with SE. PATIENTS AND METHODS: Nine patients with mean age 12.3 years and a follow up from 1 to 11 years were studied, analyzing etiology, neurological examination, psychometric evaluation, age at onset and semiology of epileptic seizures, EEGs and neuro-radiological findings, response to treatment and evolution. RESULTS: Etiologies were: porencephaly in 4 cases, hypoxic-ischemic encephalopathy in 2, bacterial meningitis in 1, herpetic encephalitis in 1 and meningo-encephalitis in the last. All cases had mental retardation. Mean age at onset of epileptic seizures was 4 years. Mean age at onset of startle seizures was 6. The startle seizures were characterized by sudden tonic contractions of the paretic hemibody, provoked by auditory stimulus in 6, somatosensory in 2 and both types of stimulus in 1. Falls were observed in 6 patients. Seizures were daily and always when awake. Unprovoked focal seizures with or without secondary generalization were found in 8 cases, and in 6 they presented previously to the startle seizures. Interictal EEGs showed unilateral spikes in 3, bilateral spikes in 3 and generalized polyspike-wave paroxysms in the other three cases. Ictal EEGs were obtained in 8 of the 9 patients and showed diffuse paroxysms of rhythms 6-11 Hz. Cerebral CT scan and/or MRI revealed extensive unilateral encephalomalacia in 5 and porencephaly in 4. The different antiepileptic schedules were unsuccessful in all cases. Surgery was performed in two patients. They are free of seizures after 1 to 4 years of follow up. CONCLUSION: SE should be considered as a distinctive epileptic syndrome or a particular electro-clinical evolution in patients with a large unilateral brain lesion associated with provoked reflex seizures usually refractory to antiepileptic drugs. Epileptic surgery should be considered. PMID- 14752710 TI - [Lesions in the pars compacta substantiae nigra and the subthalamic nucleus modify the density of muscarinic receptors in different nuclei of the basal ganglia]. AB - INTRODUCTION: Several studies that has focused to the dopaminergic transmission in the basal ganglia in parkinsonian condition, but only a few article has taking into account the imbalance between dopaminergic and cholinergic transmission. OBJECTIVE: To evaluate the muscarinic cholinergic receptors density in SNc and PPN in the 6-OHDA model. MATERIALS AND METHODS: Were organized five experimental groups in correspondence to the place of the lesion: I. Non treated rats, II. 6 OHDA lesion in SNc, III. 6-OHDA lesion in SNc + quinolinic acid lesion in NST, IV. Sham operated rats, V. Quinolinic acid in STN. Were obtained coronal sections of 20 microm thickness of SNc and PPN from rats and in these sections was evaluated the muscarinic receptors density through autoradiographic technique with [3H]quinuclidinylbenzilate (QNB) (1.23 nM). The muscarinic antagonist atropine (1 microM) was utilized as non-specific union. The density was evaluated in both hemispheres and the density optical was converted in fentomolas/mg of tissue with base to values obtained from tritium standards. RESULTS: Significant diminution of the muscarinic receptors density was found in the SNc ipsilateral to the 6-OHDA lesion from experimental groups II (t=2.76; p<0.05) and III (t=4.06; p<0.05). In the group V, was seen a significant increase of muscarinic receptor density in the SNc ipsilateral to the 6-OHDA lesion. The comparison between experimental groups evidenced significant differences among them (F=13.13; p<0.001) with a significant decrease in the density from SNc of groups II and III and significant increase in the density from SNc of group V in comparison of the others groups. In relation to PPN, muscarinic receptors density from right PPN ipsilateral to the 6-OHDA lesion, shown significant differences (F=3.93; p<0.01) between the experimental groups with a significant increase of this variable in the group II. CONCLUSIONS: These results signal a modification of cholinergic activity after 6-OHDA lesion. The changes in the muscarinic receptors populations located in SNc and PPN could be part of different compensatory mechanisms to attempt ameliorate the imbalance between dopaminergic and cholinergic transmission that it was installed after denervation of nigrostriatal forebrain bundle. The excitotoxic lesion of STN impose a new adjust mechanism for cell from PPN, which could be expressed in the changes of muscarinic cholinergic receptors population at the level of SNc. PMID- 14752711 TI - [Space-occupying cerebellar infarcts: surgical or conservative treatment?]. AB - INTRODUCTION: Space-occupying cerebellar infarcts (SOCI) make up a subgroup within the ischemic pathologies of the brain which are generally well tolerated clinically and usually respond well to medical treatment. Yet, in some cases, due to their volume and localisation, they can be accompanied by a progressive deterioration that requires surgical treatment. We report the clinical cases of two patients with SOCI that illustrate both therapeutic alternatives. CASE REPORTS: We describe the case of a 65-year-old male patient with vertiginous syndrome, inability to walk and headache. Computerised axial tomography (CAT) scanning showed a SOCI in the left hemisphere that displaced the 4th ventricle, effaced the cisterns and gave rise to the hydrocephalus. The patient improved clinically with medical treatment and a week later was able to walk unaided. We also describe the case of a 75-year-old female patient with symptoms of deterioration in the level of consciousness. CAT and magnetic resonance scans showed an extensive SOCI in the right hemisphere with displacement of the 4th ventricle, effacement of the perimesencephalic cisterns and triventricular hydrocephalus. A suboccipital craniectomy was performed, along with excision of any necrotic material and duroplasty. At 24 h the patient expressed herself coherently and followed orders. She presented ataxia and dysmetry on the right hand side following the operation, which were compensated with rehabilitating therapy. CONCLUSIONS: Our clinical case reports illustrate the alternatives available when dealing with patients with cerebellar infarct that show expanding clinical-radiological characteristics. These patients must be carefully controlled because of the risk of clinical deterioration, in which case timely surgical intervention can increase their survival. PMID- 14752712 TI - [Factor V Leiden mutation as a cause of venous thrombosis]. AB - INTRODUCTION: Venous thrombosis is infrequent in paediatrics. A lot of prothrombotic risk factors have been described. Disturbances of coagulation are present in more than half of children with stroke. Leiden V factor mutation is emphasized as one of the most common genetic cause of deep venous thrombosis in Caucasian children and adults, which represents 20-25%, depending on series. CASE REPORT: Two years-old girl with hypoxic-ischemic disease and West syndrome, which presents a deep venous thrombosis in both legs. This evolved to gangrene, requiring surgical exceresis of scar and amputation of 2nd, 3rd and 4th fingers of left foot. She also presented a right frontoparietal intraparenchima haemorrhagic stroke with tetraventricular bleeding and hydrocephalus. It required an external ventricular derivation and later a Ventricular-peritoneal valve. CONCLUSIONS: The coagulation study confirmed the Leiden V factor mutation in our patient. Later studies showed the same mutation in the father. The risk of recurrence and the severity of venous thrombosis indicate life antiaggregant treatment, currently maintained with aspirin. PMID- 14752713 TI - [A case of epilepsy simulating a pheochromocytoma]. AB - INTRODUCTION: Autonomic epilepsy is a rare entity that results from an epileptogenic focus involving certain structures belonging to the autonomous nervous system; it is characterised by the occurrence of convulsive seizures that consist in autonomic phenomena. CASE REPORT: We report the case of a 28-year-old female patient who suffered paroxysmal episodes similar to those accompanying a pheochromocytoma. Following a thorough examination, both the presence of a tumour and the anomalous production of catecholamines were excluded and, although it was not possible to provide electroencephalographic evidence, the patient was diagnosed as suffering from autonomous epilepsy, which was later confirmed by the clinical efficacy of valproic acid therapy. CONCLUSIONS: Autonomic seizures often show characteristics that are singular or similar to those of other clinical entities. This makes the disorder difficult to diagnose and sometimes requires therapeutic testing with anticonvulsive drugs. PMID- 14752714 TI - [Meningitis by Listeria in children]. AB - INTRODUCTION: Listeria monocytogenes is a rare cause of bacterial meningoencephalitis in the non-immunocompromised pediatric population. On occasions, the absence of differential characteristics with other bacteria that cause meningitis delays diagnosis and hence treatment, worsening the prognosis. CASE REPORT: We present a case of a previously healthy, non-immunocompromised teenager who was admitted to hospital with meningitis caused by Listeria. DISCUSSION: We review the literature related to this case, noting the increasing incidence of this microorganism in the etiopathogenesis of meningoencephalitis, reason for which it has to be kept in mind in the differential diagnosis at the time of admission. PMID- 14752715 TI - [Malignant middle cerebral artery infarction: medical or surgical treatment?]. AB - INTRODUCTION: Post-ischemic brain edema occurs in 10-20% of cases of infarction of the middle cerebral artery and is the main cause of early death following a completed stroke. This complication, which is known as malignant middle cerebral artery infarction (MMCI), has a mortality rate of 78% when treated medically and thus requires a different management, such as a decompressive craniectomy. The main aim of this study is to review this procedure. DEVELOPMENT: We conducted a search in the literature published over the last 20 years on this subject. Most of the studies are series of clinical cases with very favourable surgical outcomes. In non-random case-control studies the mortality rate was seen to decrease in the surgical group, and more so if the intervention was carried out early, as compared to the group that underwent medical treatment. Another study that compared decompressive craniectomy with hypothermia showed a higher survival rate in the surgical group. The post-surgery morbidity rate has not been determined, although it seems to be lower in infarction of the non-dominant hemisphere and in younger patients. CONCLUSIONS: The low degree of conclusiveness of the studies published to date only enables us to offer one practical opinion concerning this issue: decompressive craniectomy should be evaluated on an individual basis in patients with MMCI who do not respond to medical treatment. The final decision and the most appropriate moment to operate on the patient following the stroke must be based on the family's opinion and on the clinical features of the patient. PMID- 14752716 TI - [Endogenous neuronal protection: an alternative approach]. AB - INTRODUCTION: Despite a great number of neuroprotective agents have been evaluated successfully in experimental paradigms of brain ischemia and trauma, the results obtained in clinical trials in general do not satisfy the proposed criteria of efficacy. This fact leads to the searching of alternative approaches in order to orient the efforts towards neuronal protection with better perspectives. METHOD: By analyzing the brain tissue response after an ischemic insult we could observe that preconditioning with brief periods of ischemia produces tolerance, increasing the resistance against later lethal insults. Other stimuli, such as hypoxia and cortical spreading depression, induce similar responses. All of them, selectively induce specific genes, by acting collectively to increase the surviving potential of the nerve cells, promoting the functional recovery. Administration of some of these genes, or the proteins for which they codify--among them neurotrophic factors are remarkable examples--leads to interesting findings in the global and focal ischemia in rodents, which are mainly related to the inhibition of the ischemia-induced neuronal apoptosis. CONCLUSION: These findings also lead to think in an alternative approach for the neuronal protection, which take into consideration the brain endogen response as a promissory choice for a novel therapy. PMID- 14752717 TI - [Treatment of the epileptic patient in special situations]. AB - AIMS: It is known that epileptic seizures can present not only in neurological diseases but also in a number of different medical diseases, and are important in the prognosis and treatment of the primary disease. The purpose of this study is to review a series of patients with epilepsy whose therapy involves a series of peculiarities. DEVELOPMENT: We reviewed the treatment of epilepsy in groups of patients with the following characteristics: mental disability, cardiovascular disease, respiratory disease, liver failure, hepatic porphyrias, liver transplant, kidney failure, haemodialysis, kidney transplant, infectious diseases, acquired immunodeficiency syndrome, diseases of the digestive tract, endocrine diseases, preventive antiepileptic therapy, and surgical diseases. CONCLUSIONS: The increase in life expectancy, together with the ensuing growth in the number of medical diseases, surgical procedures and pharmacological treatments, means that we often have to deal with epileptic seizures in these groups of patients. Many of the epileptic seizures presented in these patients are generally of the acute symptomatic type; they therefore require no long-term antiepileptic treatment and have a good prognosis, once the situation or disease that triggers them is righted or treated. When it comes to choosing a possible antiepileptic drug for these patients, basically all the pharmacological treatments for these medical diseases must be considered. This is because of the possible interactions that may reciprocally alter the effectiveness of the two types of drugs and, similarly, their pharmacokinetics, pharmacodynamic behaviour and preferred choice of pathways. PMID- 14752718 TI - [Contributions to epilepsy genetics]. AB - INTRODUCTION AND DEVELOPMENT: In recent years the progress made in genetics has revolutionised our knowledge of the physiopathological mechanisms governing epilepsy. Just 20 years ago, none of the genes involved in epilepsy were known. Since then 11 genes have been identified and this has enabled us to better understand the mechanisms that generate the seizures. This is not only of interest to academics but can also have a practical application by helping when it comes to making a rational choice of a drug in a particular epileptic syndrome, or by allowing new drugs to be developed as new therapeutic targets are identified. CONCLUSIONS: Genetics can help us to understand the mechanisms that cause resistance to drugs and to determine the risk of a patient's suffering from idiosyncratic side effects or not. It is therefore to be expected that over the next few years it will become a fundamental body of doctrine for the daily management of epileptic patients. PMID- 14752719 TI - [Pharmacology in epilepsy. Where are we heading for?]. AB - AIMS: The aim of this study was to conduct a critical evaluation of the contribution made by the latest antiepileptic drugs (AED) and to describe the pharmacological therapeutic strategies, which do not contemplate the use of traditional AED, employed to treat pharmacoresistant patients. DEVELOPMENT: After comparing the mechanisms of action, as well as the pharmacokinetic and pharmacodynamic characteristics of the classical and new AED, and examining the advantages and disadvantages of each of them, it still turns out to be impossible to achieve a total control of seizures in 20-25% of cases. This is due to the polymorphisms of the enzymes and of their inducers, to the over-expression of the carrier proteins (PGP, MRP) and to polymorphisms undergone by the receptors. CONCLUSIONS: Although there is a need for new AED to be synthesized which can be used to attempt to reduce the rate of pharmacoresistant patients, other strategies must also be developed in the field of pharmacogenomics, and more specifically with regard to AED that are not carried by PGP or MRP. Another important area is that of substances that antagonize these carriers and, thus, allow AED to reach the site where they are to act. PMID- 14752721 TI - [The effects of epileptic seizures on sleep architecture]. AB - AIMS AND DEVELOPMENT: Many studies have appeared in the literature that aim to analyse the influence of sleep and its stages on the behaviour of seizures, but very few look into the opposite effect, that is, the effect of the seizures on the structure of sleep. The influence exerted by a number of variables, such as the presence of seizures during the waking state prior to the study, the administration or withdrawal of antiepileptic drugs, sleep disorders during the night in the laboratory or the presence of concomitant sleep pathologies, together with the difficulties involved in monitoring or controlling all these factors gives rise to varying results in the publications examined. We offer a brief review of the components of sleep and its stages together with an update of the literature on the influence seizures exert on them. CONCLUSIONS: With regard to the macrostructure of sleep, there seems to be general agreement about the fact that nocturnal generalised seizures alter the architecture of sleep, with a reduction in the efficiency of sleep and a drop in the amount of REM sleep. There also appears to be a high degree of concurrence about how recurring partial seizures on the night of the study affect the pattern of sleep, REM sleep being especially sensitive. No generalised agreement exists concerning the influence exerted by isolated partial seizures or by the seizures that take place during the waking state prior to the night the recording is made. Studies into the influence exerted by seizures on the microstructure of sleep (CAP) are scarce and are limited to specific pathologies. A deeper understanding of these interactions will provide us with key data about the clinical manifestations and the physiopathology of epileptic diseases. PMID- 14752720 TI - [Epilepsy and sleep-wake cycle]. AB - INTRODUCTION: Sleep and epilepsy have strong reciprocal influences and this is true for all kinds of epilepsies. The interictal paroxysmal activity increases in slow wave sleep and the localization of the primary epileptogenic area is more reliable in paradoxical sleep. DEVELOPMENT: From a practical point of view, an all-night sleep recording or a short-term sleep EEG recording during daytime, preceded or not by sleep deprivation, are both convenient procedures for the diagnosis and for evaluating the prognosis of epilepsy. Aldrich et al have reported the usefulness of video-EEG polysomnography in the diagnosis of nocturnal seizures versus non-epileptic manifestations such NREM/REM dyssomnias, even in patients with no known history of epilepsy. Sleepiness and nocturnal sleep disturbances have long been recognized in epileptic patients. A review of the literature shows a high incidence of sleep complaints among epileptics. The interaction of epilepsy, antiepileptic drugs and the sleep-wake cycle has been extensively investigated. PMID- 14752722 TI - [Idiopathic epilepsies: some therapeutic aspects]. AB - AIMS: The purpose of this study was to determine the therapeutic approach to be used in localisation-related and generalised epilepsies and idiopathic epileptic syndromes. DEVELOPMENT: Recent literature on the subject was reviewed, as were the records on a total of 118 patients from two paediatric neurology units between the years 2000 and 2003. With regard to the localisation-related cases, the following recommendations are made: 1. Treatment with monotherapy; 2. Low doses, since any antiepileptic drug can make epilepsy worse, and more so in the case of RBEI; 3. If the seizures get worse with treatment, the doses must be reduced instead of increased; 4) Carbamazepine (CBZ) and oxcarbazepine (OXC) are first choice drugs; clobazam (CLB) is indicated in OBEI and in some atypical BPEI, in which steroids in monotherapy can occasionally prove useful; valproate (VPA) is an alternative for cases of intolerance and exacerbation, and 5. Two year treatment and electroencephalogram (EEG) monitoring for exacerbation. As regards idiopathic generalised epilepsies: 1. VPA in monotherapy is recommended in all the forms, 48% were controlled; 18% were controlled with VPA + lamotrigine (LTG); 2. Childhood absence epilepsy is controlled up to 50% with VPA and 85% with VPA + ethosuximide (ESM); 3. LTG, CLB, topiramate (TPM) and Rivotril (CLN) are alternatives to be considered in all types of epilepsies and syndromes that are resistant to medication, and 4. In GCTS, VPA should be chosen in low doses in juvenile myoclonic epilepsy of Janz. PMID- 14752723 TI - [Whipple's disease confined to the brain. A clinical case with pathological confirmation at necropsy]. PMID- 14752724 TI - [Primary osteosarcoma of the cervical vertebrae]. PMID- 14752725 TI - [Thrombosis of the superior sagittal sinus in an infant: secondary to prolonged exposure to mesalazine?]. PMID- 14752726 TI - [Blood ammonia and transaminases in a term infant with perinatal asphyxia]. PMID- 14752728 TI - Organ and tumor motion: an overview. AB - An overview is presented of some of the issues that arise when considering how to manage motion of the patient (including setup errors), the tumor within the patient, and normal tissues that are sufficiently close to the tumor as to be likely to be at least partially irradiated. Problems arise in 3 areas: (1) at the tumor periphery where the question of just where the tumor surface is must be decided, (2) within the tumor proper where interplay effects between tumor motion and the radiation application (eg, in intensity-modulated radiation therapy) may result in dose inhomogeneity within the tumor, and (3) just outside the target volume where the extent to which organs at risk are to be irradiated (which needs to be based on an estimate of the consequences to them) must be decided. In the context of these problems, one must decide how to deal with potential motion. This involves 3 issues: (1) how to limit motion-the available techniques ranging from simple and unobtrusive to complicated, quasi-invasive, and quite costly; (2) how to handle the residual motion; and (3) how to calculate and present the consequences of the residual motion. These issues are discussed, and the importance of an explicit estimation of uncertainty is emphasized. PMID- 14752729 TI - Intrafractional tumor motion: lung and liver. AB - Three-dimensional (3D) dose distribution has been improved by 3D conformation and intensity modulation in external radiotherapy. Interfractional uncertainty has been reduced by image-guided setup techniques. Reduction of ambiguity because of intrafractional target motion is the next step forward. Respiratory organ motion is known to be the largest intrafractional organ motion. Radiotherapy techniques controlling, gating, or tracking respiratory motion are under investigation to use smaller safety margins and higher doses for moving tumors. However, data on intrafractional tumor motion are sparse. We developed a fluoroscopic real-time tracking system and implantation techniques of fiducial markers for moving organs and have been accumulating knowledge about internal tumor motion. We also found the importance of 4-dimensional treatment planning to account for tumor motion in precision radiotherapy. This article reviews the current basic knowledge on respiratory physiology and summarizes the accumulating knowledge on internal motion of lung and liver tumors. PMID- 14752730 TI - Artifacts in computed tomography scanning of moving objects. AB - Target volumes in the thorax and abdomen are commonly computed tomography (CT) scanned during light respiration. In this article, we analyze the distortions introduced in helical scanning of moving objects. Objects of known geometry are placed on a moving sled and scanned in a multirow helical CT scanner. The motion of the sled approximates the magnitude and velocity of organ movement in patients during light respiration (amplitude 1 cm, period 4 seconds). Scans of the phantom are obtained in high speed mode at incremental phases of respiration, and the resulting images are compared with scans obtained when the phantom is static. Computer simulations of the scan process are also performed to interpret the results and extend the analysis to a greater range of parameters in scanning, motion, and object size. Resulting scans show that spherical test objects can be shortened by as much as 2 cm or twice the periodic motion amplitude. Object shape was significantly distorted, and the geometric center of the object was displaced by as much as +/-0.8 cm. Computer simulation results qualitatively agree with the experimentally observed phantom images. These simulations predict that the effect is clearly observable even if the amplitude is decreased to 0.5 cm. Implications of scanning moving objects on treatment planning are discussed. PMID- 14752731 TI - Current ICRU definitions of volumes: limitations and future directions. AB - Volume definitions used in radiation treatment planning of 3-dimensional conformal radiation therapy (3DCRT) and intensity modulated radiation therapy (IMRT) play and important role in advancing these treatment modalities. IMRT is now recognized to be more sensitive to geometric uncertainties than 3DCRT and conventional radiation therapy because of its characteristic sharper dose gradients around target volumes and organs at risk. This article reviews the current state of the art in specifying volumes for 3DCRT and IMRT and discusses some of the limitations and potential pitfalls with the current International Commission on Radiation Units and Measurements (ICRU) Reports 50 and 62 recommendations, particularly in regard to issues of organ motion when irradiated by a time-dependent irradiation modality such as dynamic multileaf collimation IMRT or other methods that have a time component in beam delivery. PMID- 14752732 TI - Effects of motion on the total dose distribution. AB - The success of highly target-conformal treatments such as intensity-modulated radiotherapy (IMRT) can be compromised by motion of the inner organs and random patient setup errors. This article gives an overview of different studies that looked at the effect of organ motion and setup errors on radiation therapy dose distributions, both from a qualitative and quantitative point of view. The qualitative findings are generally applicable (ie, case independent). It is found that motion always leads to a blurring of the dose distribution. In addition, there are so-called interplay effects if the treatment delivery involves moving parts, such as multileaf collimators. After a large number of fractions, the interplay effects lead to a normal distribution of the dose value around the average blurred value. Thirdly, organ motion can also cause a spatial deformation of the dose distribution. Quantitatively it has been found that both deformation and interplay effects appear to be small (in the order of 1%-2%) in many typical clinical cases. The dominant effect is the blurring of the dose distribution, which is, in essence, independent of the treatment technique, and is not more pronounced in IMRT than in more conventional treatment techniques. However, because in IMRT there is a tendency to reduce or compromise target margins, the blurring has potentially a bigger effect on the outcome of IMRT, unless precision dose delivery techniques (such as gated or motion-synchronized beams) are used. An alternative to the use of margins is to do the planning based on blurred dose distributions. PMID- 14752733 TI - Errors and margins in radiotherapy. AB - Clinical radiotherapy procedures aim at high accuracy. However, there are many error sources that act during treatment preparation and execution that limit the accuracy. As a consequence, a safety margin is required to ensure that the planned dose is actually delivered to the target for (almost) all patients. Before treatment planning, a planning computed tomography scan is made. In particular, motion of skin with respect to the internal anatomy limits the reproducibility of this step, introducing a systematic setup error. The second important error source is organ motion. The tumor is imaged in an arbitrary position, leading to a systematic organ motion error. The image may also be distorted because of the interference of the scanning process and organ motion. A further systematic error introduced during treatment planning is caused by the delineation process. During treatment, the most important errors are setup error and organ motion leading to day-to-day variations. There are many ways to define the margins required for these errors. In this article, an overview is given of errors in radiotherapy and margin recipes, based on physical and biological considerations. Respiration motion is treated separately. PMID- 14752734 TI - Deep inspiration breath hold and respiratory gating strategies for reducing organ motion in radiation treatment. AB - We examine 2 strategies for reducing respiration-induced organ motion in radiation treatment: deep inspiration breath hold (DIBH) and respiratory gating. DIBH is a controlled breathing technique in which the patient performs a supervised breath hold during treatment. The technique offers 2 benefits: reduced respiratory motion from the breath hold and increased normal tissue sparing from the increased lung volume. In respiratory-gated treatment, a device external to the patient monitors breathing and allows delivery of radiation only during certain time intervals, synchronous with the patient's respiratory cycle. Gated treatment offers reduced respiratory motion with less patient effort than DIBH. We briefly survey the development of these 2 strategies, describe their clinical implementation for treatment of thoracic and liver tumors at the Memorial Sloan Kettering Cancer Center, and discuss their advantages and limitations. PMID- 14752735 TI - Interventions to reduce organ motion effects in radiation delivery. AB - Because organ motion during a radiation treatment fraction can be substantial, with resultant alterations in organ volume definition and dosimetry, interventions to reduce the impact of intratreatment organ motion are required for many patients if dose escalation and reduction of dose to normal tissue are treatment goals. Before radiation delivery, fluoroscopy, cine computed tomography scans and magnetic resonance imaging can be used to measure the magnitude of organ motion for an individual patient. Strategies to minimize organ motion caused by breathing during radiation include altering breathing patterns, treating during voluntary or controlled breath holds, gating the radiation beam or tracking it with organ motion. Here the first 2 interventions will be primarily discussed. Approaches to minimize nonrespiratory organ motion include maintaining the same preparative regimen before each treatment and ensuring comfortable immobilization and short overall treatment time. PMID- 14752736 TI - 4-dimensional computed tomography imaging and treatment planning. AB - In the era of conformal therapy and intensity-modulated therapy, there is an increased desire to raise tumor dose to facilitate improved survival and decrease normal tissue dose to reduce treatment-related complications. Setup accuracy and internal motion limit our ability to reduce margins. Internal motion has both interfraction and intrafraction components, although only the intrafraction component will be addressed here. Intrafraction motion is significant for lung, liver, and pancreatic radiotherapy and to a lesser extent breast and prostate radiotherapy. A method to explicitly account for intrafraction motion is to temporally adjust the treatment beam based on the tumor position with time such that the motion of the radiation beam is synchronized with the tumor motion. This addition of time into the 3-dimensional treatment process is termed 4-dimensional (4D) radiotherapy. Four-dimensional radiotherapy may allow safe clinical target volume-planning target volume margin reduction to achieve the goals of raised tumor dose and decreased normal tissue dose. This article discusses methodology for 4D CT imaging and 4D treatment planning, with some comments on 4D radiation delivery. PMID- 14752737 TI - Tracking moving organs in real time. AB - In an ideal radiotherapy procedure, the treatment system would continuously adapt the radiation beam delivery to changes in the tumor position. The development of such a tracking capability has been underway for more than 10 years, beginning with the CyberKnife image-guided radiosurgery system. In that time, much has been learned about the nature of tumor motion and the technical issues that it presents to a practical real-time tracking system. In this article, I will review the basic concepts behind existing and proposed radiotherapy beam-tracking systems, show clinical evidence of the types of movement that are encountered in real-time tracking situations, describe the corresponding technical problems and solutions, and discuss the unresolved issues in making real-time tumor tracking a practical response to tumor motion. PMID- 14752756 TI - Afraid...outside the gates. PMID- 14752757 TI - Effects of isolated Weber B fibular fractures on the tibiotalar contact area. AB - Fractures of the lateral malleolus can occur without rupture of the deltoid ligament or fracture of the medial malleolus. Controversy exists regarding the necessity of surgery on supination-external rotation stage II ankle fractures. Theoretically, as long as the medial structures are intact, the talus cannot displace enough to cause degenerative arthritis of the ankle joint. The purpose of this study was to measure changes in contact area between the tibial plafond and the talar dome with serial displacement of the distal fibula in both a lateral and a superolateral direction. Twelve cadaver lower extremities were used. Distal fibular fractures were replicated by creating an osteotomy. Displacement was accomplished with a customized apparatus that displaced and held the distal fibula in a malaligned position. Tibiotalar contact area was measured with pressure sensitive film at the following intervals of fibular displacement: 0 mm, laterally 2 mm and 4 mm, and then posteriorly and superiorly 2 mm and 4 mm. A servohydraulic testing apparatus was used to apply the same physiologic load to all limbs while measuring contact area. Key independent variables included the direction and amount of displacement of the distal fibula. Mean tibiotalar contact area decreased from baseline (no displacement) 361.1 mm2 (SD +/- 49.0) to 162.2 mm2 (SD +/- 81.3) and 82.6 mm2 (SD +/- 30.6) for 2 mm and 4 mm lateral displacement of the distal fibula respectively. With posterior/superior displacement of 2 mm and 4 mm mean tibiotalar contact decreased to 219.3 mm2 (SD +/- 56.7) and 109.2 mm2 (SD +/- 39.0), respectively. Statistical significance was found (P <.001) when comparing normal ankle alignment with displaced fractures at all levels of displacement. PMID- 14752758 TI - Biomechanical consequences of lateral column lengthening of the calcaneus: Part I. Long plantar ligament strain. AB - Lateral column lengthening of the calcaneus has been a powerful tool used to correct peritalar subluxation in symptomatic flat feet. The mechanical basis and limits for correction with this procedure are not well understood. A flatfoot model was created on 8 fresh-frozen cadaver feet by sectioning the deltoid ligament, talonavicular capsule, and spring ligament. Strain-gauge analysis of the long plantar ligament was performed (on 6 specimens) as the lateral column was sequentially lengthened from 4 to 12 mm in 2-mm increments. Results showed that only the lateral most one-third of the long plantar ligament measured positive strain during this procedure. The medial two-thirds of the long plantar ligament and plantar fascia decreased in tension and became fully lax during lengthening. Maximum tension in the long plantar ligament was measured after placing grafts 6 mm in thickness (P <.05). Larger grafts produced additional strain in the ligament, but were not significant. The authors conclude that grafts >6 mm have no additional corrective capacity without compromising the long plantar ligament. Either larger graft size or loss of the long plantar ligament could compromise the intrinsic stability of the lateral column of the foot. These findings may decrease the incidence of complications with this procedure, specifically lateral column pain, instability, and calcaneocuboid arthrosis. PMID- 14752759 TI - Enhancement of the fixation of pyrolytic carbon implants by using atomic oxygen texturing. AB - The purpose of this study was to examine the effect on the biocompatibility and biologic fixation of atomic oxygen-textured pyrolytic carbon. The implants consisted of unalloyed pyrolytic carbon rods, with half the length of the rod textured by atomic oxygen, and the other half retaining the as-deposited surface that normally occurs during the manufacturing process. The rods were implanted transcortically across the distal portion of the femurs of 6 adult male rabbits for 8 weeks. The implants were assessed mechanically by measuring the interface strength between the implant and the bone. The implant-bone interface was also examined by light microscopy. No adverse reaction to either the as-deposited or the textured pyrolytic carbon was seen. Percent bone apposition was greater for textured implants than for the as-deposited implants; however, it was not significant. The bone apposition efficiency factor, calculated by dividing the interface strength by the fraction of bone apposition, was greater for the textured implants than for the as-deposited implants. This indicates that the fixation obtained was more effective for the textured implants. The findings of this study suggest that biologic fixation of pyrolytic carbon implants can be enhanced by surface texturing by using direct exposure to atomic oxygen, without compromising its biocompatibility. PMID- 14752760 TI - Deltoid ligament integrity in lateral malleolar fractures: a comparative analysis of arthroscopic and radiographic assessments. AB - Foot and ankle surgeons often rely on the medial clear space to evaluate competency of the deep deltoid ligament when evaluating ankle fractures. This investigation assesses the integrity of the deep deltoid ligament after lateral malleolar fracture by using direct arthroscopic visualization and medial clear space separation on plain film radiographs. The objectives of this study were to test the reliability of medial clear-space separation and the Lauge-Hansen classification scheme in predicting deep deltoid rupture in displaced lateral malleolar fractures. The medial clear space was measured on injury radiographs of 40 patients with an isolated displaced lateral malleolar fracture who underwent open reduction and internal fixation. Injury radiographs were classified according to the Lauge-Hansen scheme. Direct arthroscopic visualization was used to evaluate the deep deltoid ligament under manual stress before fracture reduction. The mean preoperative medial clear space in patients with a deep deltoid rupture (n = 13) was 6.6 +/- 2.4 mm (range, 4 to 12 mm), and in patients without a deep deltoid rupture (n = 26), it was 4.0 +/- 1.0 mm (range, 2.5 to 6 mm) (P =.002, 2-sample t test). At an injury medial clear space > or =3 mm, the false positive rate for deltoid rupture was 88.5% (P =.54, Fisher's exact test). At > or =4 mm, the false positive rate was 53.6% (P =.007). All fractures were rotational injuries according to the Lauge-Hansen system. Three fractures were not classifiable; another 3 fractures showed deltoid ligament integrity opposite the expected finding. The results indicate that, in isolated displaced fractures of the lateral malleolus, radiographic widening of the medial clear space is not a reliable indicator for deep deltoid rupture. Some fractures considered stable by the Lauge-Hansen classification may require careful scrutiny to rule out deep deltoid injury. PMID- 14752761 TI - Magnetic resonance imaging and surgical correlation of peroneus brevis tears. AB - This study reviewed retrospectively preoperative magnetic resonance imaging (MRI) and intraoperative findings of 32 patients who underwent surgical treatment of longstanding peroneus brevis tendon pathology. The purpose of this study was 3 fold: 1) to determine the sensitivity and specificity of MRI diagnosis of peroneus brevis tendon tears as confirmed by surgical findings, 2) to define the prevalence of osseous and soft-tissue pathologies that coexist with peroneus brevis injuries, and 3) to compare the occurrence rates of other associated pathologies found on MRI to that of the surgical findings. MRI diagnosis of a peroneus brevis tendon tear showed 83% sensitivity and 75% specificity to intraoperative findings. Four false positive and 2 false negative cases were identified. Coexisting conditions identified by MRI included a low-lying muscle belly/peroneus quartus (44%), anterior talofibular ligament rupture (50%), flattened/hypertrophy peroneus longus tendon (56%), increased signal intensity within peroneus longus tendon (53%), and a flat/convex fibular groove (78%). These results show that peroneus brevis tears rarely present as isolated injuries and support the need for a comprehensive preoperative clinical evaluation and MRI assessment of the entire lateral ankle complex. PMID- 14752762 TI - Modified lapidus arthrodesis: rate of nonunion in 227 cases. AB - Several studies of Lapidus arthrodesis have commented on the rate of nonunion (ranging from 3.3% to 12.0%), although these figures are based on relatively small patient populations. This study retrospectively reviewed the medical records and radiographs of 211 consecutive patients (32 men, 179 women; mean age, 46.9 years) who received modified Lapidus arthrodesis for forefoot pathology in 227 feet. In all cases, the procedure was performed using joint curettage with subchondral plate preservation and screw fixation. Patients remained nonweightbearing for 6 to 8 weeks and were monitored for a minimum of 6 months postoperatively. Nonunion was seen in 12 (5.3%) of the 227 feet that underwent modified Lapidus arthrodesis. PMID- 14752763 TI - Management of displaced intraarticular calcaneal fractures by using external ring fixation, minimally invasive open reduction, and early weightbearing. AB - Twenty-three patients with 25 intraarticular fractures of the calcaneus were treated during a 7-year period with minimally invasive open reduction of the posterior facet, external ring fixation, and early weightbearing. Skeletal traction and a minimally invasive lateral approach were used to elevate the posterior facet. Percutaneous wires, which were secured to an external ring fixator, were used to stabilize the reduction. The patients were encouraged to bear weight on the first postoperative day and during the time the fixator was in place. A retrospective chart review was performed and the Maryland Foot Score was used. The patients ranged in age from 22 to 68 years (average, 43.8 years). Using the Sanders computed tomography classification, there were 17 (68%) type II, 6 (24%) type III, and 2 (8%) type IV fracture patterns. Thirty-two percent of the patients were rated excellent, 60% rated good, and 8% rated fair. Subtalar joint motion was >50% of the uninjured foot in 21 of 25 fractures. Eight of 23 patients (35%) experienced diffuse pain in the rear foot, whereas 4 (17%) complained of localized pain in the plantar heel. Few complications were observed, with the most common being superficial skin infection at a wire insertion site. No deep infections developed. The average length of the treatment period with the fixator was 6.6 weeks (range, 5 to 9 weeks). The range of follow-up was 2 to 7 years. The results with this technique indicate that it is a viable alternative to traditional methods of open reduction and internal fixation for the management of intraarticular fractures of the calcaneus. PMID- 14752764 TI - Macrodystrophia lipomatosa: a reconstructive approach to gigantism of the foot. AB - Localized gigantism poses a challenging surgical dilemma, and it may be treated with amputation. This case report documents the application of a reconstructive approach to a severe case of pedal macrodystrophia lipomatosa in a 1-year-old girl. A series of 3 surgeries were designed to reduce the length, width, height, and overall bulk of the congenitally enlarged foot. The 3 procedures debulked the foot for normal ambulation and same-size shoe wear for both feet. The resulting functional and aesthetic improvements achieved through reconstructive treatment provided a desirable alternative to amputation. PMID- 14752765 TI - Closure of central defects of the forefoot with external fixation: a case report. AB - Excision of 1 or more central rays of the foot may complicate soft tissue coverage because large defects may result. The authors presents a technique in which an Ilizarov external fixator was used to narrow the forefoot after resection of the central rays in a patient with diabetes. After external fixator application for 8 weeks, the central defect healed uneventfully and has not shown any recurrence of ulceration after 4 years of follow-up. PMID- 14752766 TI - The use of sentinel lymph node biopsy in squamous cell carcinoma of the foot: a case report. AB - Squamous cell carcinoma (SCC) is the second most common skin cancer in humans. Because the incidence of metastasis from SCC of the skin is rare, regional lymphadenectomy is generally not recommended for the patients with clinically node-negative disease. However, in patients with an intermediate and high risk of metastasis, evaluation of the lymph nodes to detect the absence of metastatic nodal disease is a difficult task. Here, we present a patient with a large SCC on the dorsum of the foot with clinically negative inguinal and popliteal lymph nodes. Intraoperative lymphatic mapping technique was used to make the decision of the inguinal node dissection. Two sentinel lymph nodes that were biopsy negative were found; therefore, only tumor excision was performed without adding complete inguinal node dissection. The defect was reconstructed with the free flap. After a 4-year carcinoma-free period, we determined that the pathology of the sentinel lymph nodes reflected that of the inguinal region. The use of selective lymphadenectomy technique in extremity SCC is very new. However, it might be useful in staging patients with SCC of the lower extremity by being able to detect absence or presence of occult metastatic nodal disease and avoid unnecessary complete inguinal node dissection. PMID- 14752767 TI - An optimal technique for ankle arthrodesis. PMID- 14752768 TI - A plantar approach for fixation of subtalar joint arthrodesis. PMID- 14752775 TI - Umbilical cord blood transplantation in adults using myeloablative and nonmyeloablative preparative regimens. AB - Unrelated umbilical cord blood (UCB) transplantation has recently been explored in an increasing number of adult patients. The relative ease of procurement and the lower-than-anticipated risk of severe acute graft-versus-host disease has made UCB transplantation an appealing alternative to bone marrow-derived hematopoietic stem cells. The use of reduced-intensity or nonmyeloablative preparative regimens to allow engraftment of UCB broadens the scope of patients who may benefit from allogeneic immunotherapy, including elderly and medically infirm patients with no matched sibling donor. This review summarizes the available data on the use of UCB as an alternative source of hematopoietic stem cell transplantation in adult patients. PMID- 14752776 TI - Will developments in allogeneic transplantation influence treatment of adult patients with sickle cell disease? AB - With improvements in the treatment of children with sickle cell disease (SCD), there has been a significant increase in the number of patients with SCD in adult hematology practice. Quality of life and life expectancy continue to be severely compromised in adult patients; hydroxyurea is the only treatment currently available that could reduce the severity and frequency of painful episodes. Allogeneic stem cell transplantation (SCT) has been offered to children with SCD as a curative option. We discuss the implications of new developments in the field of allogeneic SCT in the treatment of adult SCD patients in light of the experience derived from pediatric transplantation. These developments include innovations in the conditioning regimens, GVHD prophylaxis, and alternative donor SCT and their possible effect on adult SCD patients. Finally, we discuss a nonmyeloablative conditioning protocol for adult SCD patients and the eligibility criteria for adult SCD patients undergoing allogeneic transplantation. PMID- 14752777 TI - Effects of organ-specific loss of insulin-like growth factor-I production on murine hematopoiesis. AB - To determine whether circulating insulin-like growth factor (IGF)-I has a role in hematopoiesis, we examined hematologic parameters in mice with markedly reduced serum levels resulting from a liver-specific inactivation of the IGF-I gene. These mice have normal postnatal growth and development, suggesting that local production of IGF-I can maintain anabolic effects. Liver-specific IGF-I-deficient (LID) mice were compared with control littermates with regard to hematopoietic parameters. Spleen cellularity was decreased in the LID mice compared with control mice. Spleen myeloid progenitors, as determined by colony-forming units granulocyte/monocyte (CFU-GM) and colony-forming units-high proliferative potential (CFU-HPP), were significantly decreased in the LID mice. Immune parameters, as indicated by the absolute number of B and T cells, did not significantly differ between the knockout and control mice. In contrast to the decreased cellularity and myelopoiesis in the spleen, bone marrow cellularity was not different between the 2 groups, but the total femoral content of CFU-GM and CFU-HPP was significantly increased in the LID mice. The decrease in splenic myelopoiesis was not due to the inability of progenitors to exit the bone marrow, because CFU-GM and burst-forming units-erythroid were significantly increased in the blood of LID mice compared with normal littermates. Administration of exogenous IGF-I to the LID mice for 4 days partially restored myelopoietic parameters in the spleen. Liver production of IGF-I and, therefore, normal serum levels of this hormone, although not necessary for general organ growth and development, seems necessary for survival or transition of myeloid progenitors into the spleen. PMID- 14752778 TI - Allogeneic versus syngeneic killer splenocytes as effector cells for the induction of graft-versus-tumor effect. AB - The effect of allogeneic versus syngeneic killer cells derived from normal or severe combined immunodeficiency disease (SCID) mice was evaluated for induction of antitumor reaction in a murine model of mammary carcinoma. Tumor cells of H-2d origin were injected intravenously into H-2(d/b) mice 24 hours after total body irradiation (4 Gy). On the following day, lymphokine-activated killer (LAK) splenocytes, derived from either minor (H-2d) or major (H-2b) histocompatibility complex (MHC)-mismatched parental normal mice or MHC (H-2b)-mismatched SCID mice, were given intravenously. LAK cells of H-2d normal or SCID mice, syngeneic to the tumor, were inoculated in parallel. The results show that LAK cells derived from minor histocompatibility complex-mismatched or MHC-mismatched parental normal mice improved the probability of tumor-free survival as compared with LAK cells syngeneic to the tumor cells, but they aggravated the severity of graft-versus host disease. SCID splenocytes serving as a source of natural killer (NK) cells were expanded and activated in vitro by rIL-2 to obtain a sufficient number of DX5+ CD3- CD8- NK cells (SCID-LAK). H-2b SCID-LAK cells did not cause graft versus-host disease and significantly delayed tumor growth compared with syngeneic H-2d SCID-LAK cells, as indicated by tumor colony assays in vitro and adoptive transfer experiments. However, the graft-versus-tumor effect was not long lasting, and treated mice finally died of tumor. Our results show an advantage of allogeneic over syngeneic cell therapy for achieving a graft-versus tumor effect by rIL-2-activated T cells and NK cells. Periodic repetition of NK treatments may be required to achieve more durable antitumor effects. PMID- 14752779 TI - Persisting posttransplantation cytomegalovirus antigenemia correlates with poor lymphocyte proliferation to cytomegalovirus antigen and predicts for increased late relapse and treatment failure. AB - Numerous clinical studies link cytomegalovirus (CMV) infection with incomplete posttransplantation T-cell recovery. We hypothesized that the inability of transplant recipients to handle CMV reactivation might correlate with a defective graft-versus-leukemia response and increased posttransplantation morbidity. Between May 1995 and August 2001, 82 patients who were CMV seropositive and survived the first 100 days after transplantation were identified for a day 100 landmark analysis of the effect of CMV reactivation patterns on eventual transplantation outcome. All patients underwent a myeloablative HLA-identical sibling donor T cell-depleted stem cell transplantation with scheduled donor T cell add-back on day 45. Median follow-up was 1032 days. Forty-two patients who had either no reactivation or only 1 positive test with quick clearance were designated as a CMV immune competent group. Forty patients designated as CMV immune deficient (ID) had at least 2 positive tests. Apart from younger age (33 versus 38 years; P =.05) in the ID group, the 2 groups were balanced for clinical characteristics. In multivariate analysis, ID patients had a significantly higher incidence of leukemia relapse (58% versus 21%; P =.03) and worse disease-free survival (31% versus 66%; P =.04). There was no significant difference in week 1 to 14 posttransplantation lymphocyte counts between the 2 groups. In 67 patients tested 3 to 6 months after transplantation, a proliferative response to CMV antigen (stimulation index > or =2) occurred in 27 of 36 immune competent patients compared with 15 of 31 ID patients (P =.006). These results show that recurrent CMV reactivation in the first 100 days after transplantation predicts for reduced disease-free survival and increased leukemic relapse beyond 100 days and correlates with inferior proliferative responses to CMV. The higher relapse rate may reflect poor immune reconstitution in ID patients or an adverse effect of prolonged antiviral treatment. PMID- 14752780 TI - Prolonged outbreak of human parainfluenza virus 3 infection in a stem cell transplant outpatient department: insights from molecular epidemiologic analysis. AB - Human parainfluenza virus type 3 (hPIV3) infections cause considerable morbidity and mortality after stem cell transplantation, and inpatient nosocomial outbreaks are common. From September 1998 to July 1999, 93 stem cell transplantation recipients at our institution contracted hPIV3, of which 66 (71%) were being followed up in our outpatient department (OPD). The peak incidence was in September and October, when 39 cases were identified; thereafter, hPIV3 incidence decreased to approximately 5 cases per month. Nucleotide sequences (778 nucleotides from variable regions of the hemagglutinin-neuraminidase gene) from 46 patient and 8 community hPIV3 isolates were compared to determine epidemiologic relatedness. Sequence analysis of OPD isolates revealed that 18 of 19 isolates from September and October and 11 of 15 isolates from November 1998 to July 1999 were genetically similar. In contrast, 2 of 3 community isolates from September and October and 0 of 5 from November to July were linked to this cluster. Symptomatic surveillance and isolation were ineffective in terminating the outbreak, suggesting asymptomatic shedding among patients, staff, or visitors or viral persistence on environmental surfaces as possible explanations. The concept of nosocomial transmission should be expanded to include the OPD for immunosuppressed patients. PMID- 14752788 TI - Recent report in the etiology of childhood cancer: 'greatest hits'. PMID- 14752781 TI - Development of early neutropenic fever, with or without bacterial infection, is still a significant complication after reduced-intensity stem cell transplantation. AB - Little information is available on the clinical characteristics of infectious complications that occur in the early period after reduced-intensity stem cell transplantation (RIST). We retrospectively investigated the clinical features of neutropenic fever and infectious episodes within 30 days after RIST in 76 patients who had received fluoroquinolones as part of their antibacterial prophylaxis. Preparative regimens included cladribine 0.66 mg/kg or fludarabine 180 mg/m2 plus busulfan 8 mg/kg. All but 1 patient survived 30 days after transplantation, and 75 patients (99%) became neutropenic within a median duration of 9 days. Neutropenic fever was observed in 29 patients (38%), and bacterial infection was confirmed in 15 (20%) of these, including bacteremia (n = 13), bacteremia plus pneumonia (n = 1), and urinary tract infection (n = 1). The causative organisms were gram-positive (n = 9) and gram-negative organisms (n = 7), with a mortality rate of 6%. Neither viral nor fungal infection was documented. Multivariate analysis showed that the presence of neutropenia at the initiation of preparative regimens was an independent risk factor for subsequent documented bacterial infections (P =.026; 95% confidence interval, 1.25-35.1). We conclude that neutropenic fever and bacteremia remain common complications in RIST. PMID- 14752789 TI - Outcome of children with high-risk acute lymphoblastic leukemia (HR-ALL): Nordic results on an intensive regimen with restricted central nervous system irradiation. AB - BACKGROUND: Improvement in outcome of childhood high-risk (HR) ALL was sought with a very intensive Nordic protocol leaving most patients without CNS-RT. METHODS: A total of 426 consecutive children entered the NOPHO-92 HR-ALL program. HR criteria included WBC > or =50 x 10(9)/L, CNS or testicular involvement, T cell, lymphomatous features, t(9;22), t(4;11), or slow response. Of these, 152 children had very high risk (VHR) with special definitions. CNS consolidation was based on high-dose MTX (8 g/m2) and ARA-C (12 g/m2) alternating. VHR patients also received cranial RT. RESULTS: The 9-year EFS was 61 +/- 3%, OS 74 +/- 2%, and EFS for T-ALL 62 +/- 4%. Cumulative incidence of isolated CNS relapse was 4.7 +/- 1%, and CNS relapse in total 9.9 +/- 2%. Poor prognostic factors were WBC > or =200 x 10(9)/L and a very slow response. CONCLUSIONS: HR-ALL was successfully treated on the NOPHO-92 regimen, with a relatively low CNS relapse rate for non irradiated children. WBC > or =200 x 10(9)/L and very slow response emerged as strong poor prognostic factors. PMID- 14752790 TI - CNS-directed therapy in young children with T-lineage acute lymphoblastic leukemia: High-dose methotrexate versus cranial irradiation. AB - BACKGROUND: Prophylactic use of cranial radiation therapy (CRT) in young children with acute lymphoblastic leukemia (ALL) is associated with significant long-term morbidity. Therefore, current treatment protocols for pediatric B-precursor ALL have abandoned prophylactic CRT in favor of intrathecal chemotherapy, combined with either high-dose methotrexate infusions (HD-MTX) or intensive systemic chemotherapy. In contrast, prophylactic CRT continues to be used in children with T-lineage ALL (T-ALL), who historically have had an inferior prognosis. We conducted a retrospective cohort study to determine the effect on survival of substituting HD-MTX for CRT in young children with T-ALL, a group that faces a high risk of long-term sequelae from CRT. PROCEDURE: Twenty-six children, diagnosed with T-ALL between the ages of 1 and 5 years, were treated on the same high-risk leukemia protocol. Central nervous system (CNS) directed therapy consisted of either CRT (1,800 cGy) or HD-MTX (three doses of 8 g/m2), depending on the treatment era in which patients were diagnosed. RESULTS: Of the 24 patients who entered remission, 12 received CRT and 12 received HD-MTX. Five-year event-free survival (EFS) (+/-SE) was 92 +/- 8% in the HD-MTX group versus 75 +/- 13% in the CRT group (P=0.23). Five-year overall survival (OS) was 100% in the HD MTX group versus 75 +/- 13% in the CRT group (P=0.07). There were no CNS recurrences in the HD-MTX group. One patient treated with CRT developed a brain tumor. CONCLUSIONS: The use of HD-MTX instead of CRT as CNS-directed therapy in very young children with T-ALL does not compromise survival, while avoiding the adverse long-term effects of cranial irradiation. PMID- 14752791 TI - Favorable outcome in 20-year follow-up of children with very-low-risk ALL and minimal standard therapy, with special reference to TEL-AML1 fusion. AB - BACKGROUND: As the treatment results of childhood ALL have improved, avoidance of late effects has become increasingly important. Identification of favorable prognostic factors helps to achieve this goal. PROCEDURE: We studied the prognostic value of TEL-AML1 fusion and a risk factor composed of age, white blood cell count (WBC), lymphomatous features, and hemoglobin level (Hb). We also compared outcome between two cohorts; cohort 1 (n=100) diagnosed 1975-1981, and cohort 2 (n=102) 1989-1991. Both cohorts were retrospectively divided in two groups: very-low-risk (WBC <10 x 10(9)/L, age 2 to <10 years, no lymphomatous features, Hb <90 g/L), and non-low-risk (=the remainder). We performed fluorescent in situ hybridization (FISH) of TEL-AML1 fusion of the marrow samples obtained at diagnosis. RESULTS: The median follow-up is 20 years in cohort 1, and 8 years in cohort 2. In both cohorts, the very-low-risk category comprised one fourth of the children. TEL-AML1 fusion was more frequent in the very-low-risk (35%) than in the non-low-risk group (17%) (P=0.03). The 8-year event-free survival (EFS) of children with the fusion was better than of those without, 74 vs. 54% (P=0.040). The 8-year EFS in the very-low-risk group was 76% in cohort 1, and 79% in cohort 2 (n.s.). In the non-low-risk groups, EFS was 39 vs. 64% (P=0.02), respectively. CONCLUSIONS: Our data support the reported association of TEL-AML1 fusion with a favorable outcome although the risk group had a greater impact. These very-long-term follow-up data also indicate that children with very low-risk ALL (slow disease) had a favorable outcome already in the late 1970s, and may be over treated with the contemporary ALL protocols. PMID- 14752792 TI - Immune responses to influenza immunization in children receiving maintenance chemotherapy for acute lymphoblastic leukemia. AB - PURPOSE: To compare the immune responses to influenza vaccine in children with acute lymphoblastic leukemia (ALL) receiving maintenance chemotherapy with those in healthy children. METHODS: Hemagglutinin-inhibition (HAI) antibody titers were determined before and after influenza vaccination in children with ALL and healthy controls. Immune responses were measured as geometric mean titers (GMT) and 4-fold rises in HAI titers. RESULTS: Although post-immunization GMT were lower in children with ALL compared to healthy children for the H1N1 antigen (P<0.001), the H3N2 antigen (P=0.03), and for the influenza B antigen (P=0.003), at least 60% of children with ALL had at least a 4-fold rise in HAI titers to each of the influenza antigens. CONCLUSIONS: While the GMT after trivalent influenza immunization in children with ALL were significantly lower than those seen in healthy children, the majority of children with ALL had 4-fold rises in HAI titers. Children receiving maintenance chemotherapy for ALL should receive yearly influenza vaccine. PMID- 14752793 TI - An examination of outcome measures to assess functional mobility in childhood survivors of osteosarcoma. AB - BACKGROUND: The purpose of this study is to examine outcome measures that assess functional mobility in adolescent survivors of osteosarcoma. We will also identify relationships between functional mobility measurements and specific measure(s) of health-related quality of life (HRQL). METHODS: Eighteen adolescent survivors of osteosarcoma at least 2 years from surgery at the Children's Hospital of Philadelphia (CHOP) participated in this study. Functional mobility, endurance, perceived exertion, and locomotor efficiency is examined with the following measurement tools: timed up and go (TUG), timed up and down stairs (TUDS), 9-min run-walk, rate of perceived exertion (RPE), and physiological cost index (PCI). The currently used evaluation of function from the American Musculoskeletal Tumor Society (MSTS) is also administered. The Short Form 36 (SF 36) measuring HRQL is used. RESULTS: MSTS scores did not correlate with quantitative functional outcome measures. MSTS scores did not correlate with the HRQL as measured by the SF-36. In contrast, subjects with a higher rate of vitality on the SF-36 walked with more efficiency, farther distance during the 9 min run-walk, and performed the TUDS and TUG with a faster speed. CONCLUSIONS: The MSTS does not correlate with actual measures of function or with HRQL. These data demonstrate the need for a new measure to determine functional mobility in survivors of osteosarcoma. PMID- 14752794 TI - The patterns of relapse in osteosarcoma: the Memorial Sloan-Kettering experience. AB - BACKGROUND: With the introduction of intensive high-dose chemotherapy for the treatment of osteosarcoma, changes in the pattern of metastases observed at relapse have been reported. To further investigate this hypothesis, the relapse patterns among groups receiving chemotherapy regimens of variable intensity at Memorial Sloan-Kettering Cancer Center were analyzed. PROCEDURE: All patients treated with the protocols T4, T5, T7, T10, and T12 were included. Patients were divided into two groups, one including those patients treated with less intense therapy (T4, T5, and T7) and the other, those treated with current regimens (T10 and T12). RESULTS: Of the 25 patients who relapsed on the earlier protocols, 76% relapsed to the lungs, 8% had local recurrences, and 16% distant metastases to the bone. The median time to first relapse was 12 months. Of the 69 patients who relapsed on the T10 and T12 protocols, 75% relapsed in the lungs, 9% had local recurrences, and 16% distant bone metastases. The median time to first relapse was 17 months. There was no statistically significant difference in the timing of relapse between the two groups studied, although a longer median time to relapse was observed for patients treated on the later protocols. The range of time to relapse was also wider in the later protocols. CONCLUSIONS: These data do not support the hypothesis that patterns of relapse are changing with alterations in osteosarcoma treatment. This limited single institutional experience can be explored further in the context of a multi-institutional effort. PMID- 14752795 TI - Methotrexate pharmacokinetics and survival in osteosarcoma. AB - BACKGROUND: The aim of this study was to analyze the relationship between exposure to high-dose methotrexate (HDMTX) and tumor response in terms of survival in children with osteosarcoma. PROCEDURE: This study included 44 patients (479 courses) who received a median dose of 5.92 g/m2 of MTX (interquartile range (IQR) 2.37 g/m2) in a 4-hr infusion. The mean area under the concentration-time curve (AUC) estimated by parametric methods (non-parametric expectation maximization, NPEM), and the mean concentration at the end of the infusion were considered to be the exposure parameters. Tumor response was recorded as disease-free survival (DFS), overall survival (OS), and histologic tumor response. The relationship between MTX exposure and survival parameters was analyzed by Cox regression. RESULTS: The group of 11 patients who were the least exposed to MTX (AUC <2,400 micromol/L hr) presented a high DFS, probably due to the shorter interval of time between MTX courses that led to a higher dose density. In patients with AUC >2,400 micromol/L hr, an increase in the AUC was related to an increase in the DFS. Significant differences were observed in the DFS between patients whose mean AUC was below or above 4,000 micromol/L hr (P=0.024), such that 4,000 micromol/L hr was considered as the minimum AUC to be aimed at for future patients. CONCLUSIONS: Dose density seems to be an important factor in osteosarcoma response, but this must be confirmed in further studies. In order to improve the response to osteosarcoma in children, it is recommended that the dose of MTX to be increased such as to obtain an AUC higher than 4,000 micromol/L hr. PMID- 14752796 TI - Stature of young people with malignant bone tumors. AB - BACKGROUND: Little is known about the aetiology of primary bone tumours. There have been conflicting reports relating to stature in young people with bone cancer. PATIENTS: We analysed height data at diagnosis for 364 patients with osteosarcoma and 356 patients with Ewing sarcoma registered on clinical trials run by the Medical Research Council (MRC) and the United Kingdom Children's Cancer Study Group (UKCCSG). MAIN OUTCOME MEASURES: Height at diagnosis for each patient was standardised for age and sex compared to national reference data with a standard deviation score (SDS) calculated for each subject. RESULTS: Those with osteosarcoma were significantly taller than the general population (mean height SDS 0.2, P=0.001). Patients with osteosarcoma of the femur were significantly taller than patients with other primary sites (mean height SDS 0.45 vs. -0.06, P=0.0001). Overall those with Ewing sarcoma were not significantly taller than the general population (mean height SDS 0.09, P=0.1), but children presenting under 15 years were taller (SDS 0.2, P=0.004) whilst older patients were not (SDS -0.07, P=0.4). In both osteosarcoma and Ewing sarcoma the mean age at diagnosis for females was significantly younger than for males. CONCLUSIONS: This study suggests that tall stature and an earlier pubertal growth spurt may be important factors in the aetiology of both osteosarcoma and Ewing sarcoma. PMID- 14752797 TI - Age is an independent prognostic factor in rhabdomyosarcoma: a report from the Soft Tissue Sarcoma Committee of the Children's Oncology Group. AB - BACKGROUND: Although age <1 year at diagnosis has been associated with a worse prognosis in rhabdomyosarcoma (RMS), the relationship of age at diagnosis to clinical presentation and outcome has not been evaluated carefully. We reviewed data from recent Intergroup Rhabdomyosarcoma Study Committee (later called Group, IRSG) trials to examine this relationship in order to estimate prognosis more accurately and further refine treatment. PROCEDURE: We used data from IRS-III, IV Pilot, and -IV (1983-97, N=2,343) to study the relationship of patient age with clinical features and prognosis in a large cohort of patients treated with contemporary therapy. RESULTS: We showed that, after adjusting for important prognostic factors, age was an independent risk factor for treatment failure and patients could be classified into three failure-risk categories based on age (i.e., <1 year; 1-9 years; >10 years). Infants and adolescents were more likely to have unfavorable features, including alveolar or undifferentiated tumors and advanced Group and Stage, and also had significantly poorer failure-free survival (FFS) than did children aged 1-9 (53 and 51% vs. 72%, P<0.001). Although there was a difference in FFS among age categories, there was no evidence that age influences outcome within the three categories. CONCLUSIONS: Since age relates independently to outcome after adjustment for known risk factors, it is likely that other factors, including perhaps patients' tolerance of protocol-specified therapy, explain this relationship. PMID- 14752798 TI - Liver transplantation for hepatoblastoma: results from the International Society of Pediatric Oncology (SIOP) study SIOPEL-1 and review of the world experience. AB - BACKGROUND: For hepatoblastoma (HB) that remains unresectable by partial hepatectomy after chemotherapy, total hepatectomy with orthotopic liver transplantation (LTX) has been advocated as the best treatment option. The role of LTX in the overall management of HB is still, however, unclear. PROCEDURE: The results of LTX from the first study of HB by the International Society of Pediatric Oncology, SIOPEL-1, were analyzed. In addition, the world experience of LTX for HB was extensively reviewed. Twelve patients in the SIOPEL-1 study underwent a LTX. Median (range) follow-up at Dec. 31, 2001 was 117 months (52 125) since LTX. RESULTS: Overall survival at 10 years post-LTX was 85% for the seven children who received a "primary LTX" and 40% for the 5 children who underwent a "rescue LTX" after previous partial hepatectomy. In the world experience (147 cases), the overall survival rate at 6 year post-LTX was 82% for 106 patients who received a "primary LTX" and 30% for 41 patients who underwent a "rescue LTX." Multivariate analysis of patients undergoing primary LTX showed that only macroscopic venous invasion had a significant impact (P-value: 0.045 with a hazard ratio of 2.96) on overall survival. CONCLUSIONS: Orthotopic LTX has added a new dimension to the treatment of HB unresectable by partial hepatectomy. Because of the rarity of the disease and to optimize results, children with extensive HB should be treated in centers with surgical expertise in pediatric major liver resection and LTX, in close collaboration with pediatric oncologists, radiologists, and histopathologists. PMID- 14752799 TI - A prospective study of thyroid nodular disease in children and adolescents in western Poland from 1996 to 2000 and the incidence of thyroid carcinoma relative to iodine deficiency and the Chernobyl disaster. AB - BACKGROUND: In Poland, where soil is deficient in iodine, supplementation of iodine was introduced in 1935, discontinued in 1980, and then re-introduced in 1997. One of the effects of inadequate iodine intake, prior to 1997, was an increase in the prevalence of thyroid nodular disease (TND) in children. Chernobyl, located in the neighbouring country of Ukraine, suffered a catastrophic nuclear explosion in April 1986. PROCEDURE: A total of 411 children with TND (an incidence of 7.53/100,000) were diagnosed and registered in western Poland between 1996 and 2000 and further evaluated as a population-based study. RESULTS: Based on the patient's clinical status, ultrasound examination, scintiscan, laboratory tests, cytology and the family history, many of the patients qualified for surgery and, as a result, histopathologic data were obtained from 155 of the 411 patients operated on to date (37.7% of all TND). Thyroid carcinoma was detected in 37 of the operated children, i.e. 23.9% or 9.0% of all children with TND with a median incidence of 0.68/100,000. Papillary carcinoma was the predominant histologic type (26-70.3%) compared to follicular carcinoma (10-27.0%) and medullary carcinoma (1-2.7%). Retrospective analysis of the figures for the 23 years (1972-1995) showed that in that period a total of 23 thyroid carcinomas were registered. However, only 12 of these were detected in the 20-year period between 1972 and 1991, none in the years 1992-1993 and, significantly, 11 from 1994 to 1995. CONCLUSIONS: Thyroid carcinoma appears to be an ongoing and increasing problem in the children and adolescents of our region, and it is developing more intensively when compared, both to other parts of Poland and to previous statistics (2000 vs. 1985; P<0.002). Iodine deficiency and radiation resulting from the Chernobyl disaster might be important risk factors in the development of thyroid carcinoma in the young population analysed in our region in the period since 1994. The high percentage of follicular carcinoma and follicular adenoma with an undetermined prognosis (19 out of 46) indicates that the long-term iodine deficiency in our region may be more significant in the pathogenesis of malignant transformation than has previously been postulated. PMID- 14752800 TI - Phase I study of oral cyclophosphamide and oral topotecan for children with recurrent or refractory solid tumors. AB - BACKGROUND: To determine the maximum-tolerated duration and dose-limiting toxicity of a daily schedule of orally administered cyclophosphamide and topotecan in pediatric patients with recurrent or refractory malignant solid tumors. METHODS: Patients received oral cyclophosphamide (50 mg/m2/dose) in the morning followed by topotecan (0.8 mg/m2/dose) 8-12 hr later for an escalating number of consecutive days (10, 14, and 17 days). RESULTS: Seventeen pediatric patients were treated with oral cyclophosphamide and topotecan for durations of 10-17 days for a total of 58 treatment courses. Reversible hematologic toxicity (neutropenia and thrombocytopenia) was the dose-limiting toxicity. Nonhematologic toxicities of greater than grade 3 were not observed. A partial response (neuroblastoma following myeloablative chemotherapy and stem cell rescue) and prolonged stable disease (medulloblastoma) were each observed in one patient. CONCLUSIONS: The recommended duration of therapy with a daily schedule of both oral cyclophosphamide (50 mg/m2/day) and topotecan (0.8 mg/m2/day) for previously treated pediatric patients with recurrent or refractory solid tumors is 14 consecutive days. The observed dose limiting toxicity (DLT) was reversible neutropenia. This regimen was well tolerated in heavily pretreated patients and demonstrated activity against recurrent pediatric solid tumors. PMID- 14752801 TI - Genetic epidemiology of neuroblastoma: a study of 426 cases at the Institut Gustave-Roussy in France. AB - BACKGROUND: Genetic susceptibility to neuroblastoma (NB) is now highly probable and the likelihood that it may also confer a higher risk of other cancer types has been suggested. The aim of this study was to estimate the fraction of inherited cases and penetrance associated with a carrier status, and to investigate the hypothesis that susceptibility cancer genes might be non specific. PROCEDURE: The family history of 426 children treated for NB at the Institut Gustave Roussy was obtained. The excess of relatives affected by NB or early-onset cancer (EOC) was sought using the standardized incidence ratio (SIR). The risk of NB among sibs was estimated taking into account the age of patients' sibs. Estimation of penetrance in hereditary cases and of the proportion of sporadic cases was obtained using segregation analysis of proband sibships. RESULTS: There was a positive family history of NB or ganglioneuroma in 5 of the 426 cases (1.2%). A highly significant excess of NB was found among relatives (SIR=11.4, 95% CI: 3.7-26.5). The excess of EOC (SIR=1.22, 95% CI=0.92-1.58) was non-significant, but it was of borderline significance among first-degree relatives (SIR=1.70, 95% CI=0.99-2.72). The risk of NB among sibs was estimated at 0.2% (95% CI=0.004-1.0%). Penetrance in hereditary cases was estimated at 11.4% and the proportion of inherited cases, 3.5%. CONCLUSIONS: The genetic factors heightening susceptibility to NB are most probably dominantly inherited with low penetrance and are involved in only a very small fraction of NB patients. The overall risk in sibs is very low and this should reassure parents with regard to their other children. We found some arguments for the existence of non-specific genetic susceptibility, which would slightly in crease the probability of developing any cancer. PMID- 14752802 TI - Non-conventional therapies in childhood cancer: guidelines for distinguishing non harmful from harmful therapies: a report of the SIOP Working Committee on Psychosocial Issues in Pediatric Oncology. AB - This is the 11th official document of the SIOP Working Committee on Psychosocial Issues in Pediatric Oncology, instituted in 1991. There is a tendency for some physicians to make blanket statements against the use of non-proven, non conventional therapies, even when these therapies are not harmful. There is an equal and opposite tendency on the part of many parents to do all that they possibly can for their children, including using any non-conventional therapy they feel might do some good. The health care team must open a healthy dialogue with parents that will lead to a clear distinction between those complementary therapies that are harmful and those that are not, indeed, might even be helpful psychologically if not therapeutically. PMID- 14752803 TI - Clinical features and treatment outcomes of 79 infants with immune thrombocytopenic purpura. AB - BACKGROUND: To determine the clinical features and treatment outcomes of infants with immune thrombocytopenic purpura (ITP). METHODS: Retrospective analysis of 79 infant ITP patients treated from 1987 to 2002. The data abstracted comprised age, gender, clinical features, and treatment outcomes. A score test for the trend in the odds ratios was used to determine the risk of chronic ITP with advancing age. The infants were compared to a group of contemporaneous older children with regard to bleeding severity and incidence of chronic ITP. RESULTS: The 34 female and 45 male infants had a median age of 16 months. Seventy-four presented with purpura, four with viral illnesses, and one was asymptomatic. Eight percent had active mucosal bleeding. The median platelet count was 8,000/microl. Forty infants received intravenous immunoglobulin, nine intravenous anti-D immunoglobulin, six steroids, and seven were observed without treatment. Fifty five (76%) responded to a single course of treatment. Only 9% of infants developed chronic ITP compared to 18% of children between the ages of 25 and 119 months and 47% of children 120 months or older (P<0.0005). CONCLUSIONS: Infants with ITP respond favorably to treatment and are less likely to develop chronic ITP compared to older children. PMID- 14752804 TI - Ciliary reorientation is evoked by a rise in calcium level over the entire cilium. AB - Internal Ca2+ levels control the pattern of ciliary and flagellar beating in eukaryotes. In ciliates, ciliary reversal is induced by a rise in intra-ciliary Ca2+, but the mechanism by which Ca2+ induces reversal is not known. We injected the fluorescent Ca2+ indicator Calcium Green into a ciliate Didinium nasutum and observed the intra-ciliary Ca2+ level during the initial reversed stroke preceding spontaneous cyclic reversed beating. In D. nasutum, Ca2+ rose throughout the length of the cilia undergoing initial reversed stroke. Electron microscopy with a combined oxalate-pyroantimonate method showed Ca2+ deposits distributed throughout the reversed cilia. We injected caged Ca2+ into D. nasutum and irradiated the base or mid region of the cilia with UV to locally increase Ca2+ concentration. Uncaging Ca2+ in the middle of the cilia produced reversal distally, but not proximally to the site of Ca2+ release. These results strongly suggest that not only Ca2+ influx sites, but also Ca2+ binding sites and vectoral bending machineries for ciliary reversal, are distributed throughout the cilium. PMID- 14752805 TI - Measurement of barbed ends, actin polymerization, and motility in live carcinoma cells after growth factor stimulation. AB - Motility is associated with the ability to extend F-actin-rich protrusions and depends on free barbed ends as new actin polymerization sites. To understand the function and regulation of different proteins involved in the process of generating barbed ends, e.g., cofilin and Arp2/3, fixed cell approaches have been used to determine the relative barbed end concentration in cells. The major disadvantages of these approaches are permeabilization and fixation of cells. In this work, we describe a new live-cell time-lapse microscopy assay to determine the increase of barbed ends after cell stimulation that does not use permeabilization and provides a better time resolution. We established a metastatic carcinoma cell line (MTLn3) stably expressing GFP-beta-actin at physiological levels. Stimulation of MTLn3 cells with epidermal growth factor (EGF) causes rapid and transient lamellipod protrusion along with an increase in actin polymerization at the leading edge, which can be followed in live cell experiments. By measuring the increase of F-actin at the leading edge vs. time, we were able to determine the relative increase of barbed ends after stimulation with a high temporal resolution. The F-actin as well as the barbed end concentration agrees well with published data for this cell line. Using this newly developed assay, a decrease in lamellipod extension and a large reduction of barbed ends was documented after microinjecting an anti-cofilin function blocking antibody. This assay has a high potential for applications where rapid changes in the dynamic filament population are to be measured. PMID- 14752806 TI - A subclass of myosin XI is associated with mitochondria, plastids, and the molecular chaperone subunit TCP-1alpha in maize. AB - The role and regulation of specific plant myosins in cyclosis is not well understood. In the present report, an affinity-purified antibody generated against a conserved tail region of some class XI plant myosin isoforms was used for biochemical and immunofluorescence studies of Zea mays. Myosin XI co localized with plastids and mitochondria but not with nuclei, the Golgi apparatus, endoplasmic reticulum, or peroxisomes. This suggests that myosin XI is involved in the motility of specific organelles. Myosin XI was more than 50% co localized with tailless complex polypeptide-1alpha (TCP-1alpha) in tissue sections of mature tissues located more than 1.0 mm from the apex, and the two proteins co-eluted from gel filtration and ion exchange columns. On Western blots, TCP-1alpha isoforms showed a developmental shift from the youngest 5.0 mm of the root to more mature regions that were more than 10.0 mm from the apex. This developmental shift coincided with a higher percentage of myosin XI /TCP 1alpha co-localization, and faster degradation of myosin XI by serine protease. Our results suggest that class XI plant myosin requires TCP-1alpha for regulating folding or providing protection against denaturation. PMID- 14752807 TI - The Roadblock light chains are ubiquitous components of cytoplasmic dynein that form homo- and heterodimers. AB - The Roadblock/LC7 class of light chains associate with the intermediate chains at the base of the soluble dynein particle. In mammals, there are two Roadblock isoforms (Robl1 and Robl2), one of which (Robl2) is differentially expressed in a tissue-dependent manner and is especially prominent in testis. Here we define the alpha helical content of Robl and demonstrate using both the yeast two-hybrid system and in vitro biochemistry that Robl1 and Robl2 are capable of forming homo and heterodimers. This is the first report of heterodimer formation by any cytoplasmic dynein component, and it further enlarges the number of potential cytoplasmic dynein isoforms available for binding specific cellular cargoes. In addition, we have generated an antibody that specifically recognizes Robl light chains and shows a 5-10 fold preference for Robl2 over Robl1. Using this antibody, we show that Robl is a ubiquitous cytoplasmic dynein component, being found in samples purified from brain, liver, kidney, and testis. Immunofluorescence analysis reveals that Robl is present in punctate organelles in rat neuroblastoma cells. In testis, Robl is found in Leydig cells, spermatocytes, and sperm flagella. PMID- 14752808 TI - Microtubules become more dynamic but not shorter during preprophase band formation: a possible "search-and-capture" mechanism for microtubule translocation. AB - The dynamic behavior of the microtubule cytoskeleton plays a crucial role in cellular organization, but the physical mechanisms underlying microtubule (re)organization in plant cells are poorly understood. We investigated microtubule dynamics in tobacco BY-2 suspension cells during interphase and during the formation of the preprophase band (PPB), the cytoskeletal structure that defines the site of cytokinesis. Here we show that after 2 h of microtubule accumulation in the PPB and concurrent disappearance elsewhere in the cortex, the PPB is completed and starts to breakdown exponentially already 20 min before the onset of prometaphase. During formation of the PPB, the dynamic instability, i.e., the stochastic alternating between growing and shrinking phases, of the cortical microtubules outside the PPB increases significantly, but the microtubules do not become shorter. Based on this, as well as on the cross linking of microtubules in the PPB and the lack of evidence for motor involvement, we propose a "search-and-capture" mechanism for PPB formation, in which the regulation of dynamic instability causes the cortical microtubules to become more dynamic and possibly longer, while the microtubule cross-linking activity of the developing PPB preferentially stabilizes these "searching" microtubules. Thus, microtubules gradually disappear from the cortex outside the PPB and aggregate to the forming PPB. PMID- 14752809 TI - Changes in locomotor and foraging skills in captive-born, reintroduced golden lion tamarins (Leontopithecus rosalia rosalia). AB - The behavior of reintroduced, captive-born animals is understudied, limiting the scientific understanding and utility of reintroduction as a conservation tool. This work describes changes in locomotor and foraging behaviors in captive-born golden lion tamarins over the first 18 months after their release into the wild. The subjects included 73 individuals living in and around the Poco das Antas Biological Reserve in Brazil between 1984 and 1996. The differences between animals that survived 6 months after release and those that did not indicate that initial deficiencies in locomotor and foraging abilities are related to survival. Behavioral changes in both juvenile and adult individuals during the first 6 and 18 months after release appear to be primarily related to locomotor abilities; however, the effect of provisioning on foraging abilities is unknown. Juvenile animals showed a larger number of changes relative to adults during the first 6 and 18 months, suggesting that placing tamarins into complex environments early in development may promote the expression of natural behaviors and increase survival opportunities after their release. However, when this is not possible, the best mechanism for reintroducing adult members of this species involves intensive post-release support rather than pre-release training, which confers few behavioral advantages. Recommendations for future reintroductions with this and other species include introducing animals to complex environments early in development, and collecting data systematically. PMID- 14752810 TI - Reconciliation and consolation in captive bonobos (Pan paniscus). AB - Although reconciliation in bonobos (Pan paniscus) has previously been described, it has not been analyzed heretofore by the postconflict (PC) match-control (MC) method. Furthermore, although reconciliation has been investigated before in this species, consolation has not. In this study we analyzed agonistic and affiliative contacts in all sex-class combinations to clarify and reevaluate the occurrence of reconciliation in bonobos via the PC-MC method. We also investigated the occurrence of consolation by analyzing the victims' triadic contact tendency (TCT), the influence of the sex of victims, and the relative occurrence of consolation and reconciliation. We collected 167 pairs of PC-MC observations in a captive group of bonobos (in Apeldoorn, The Netherlands). The conciliatory tendency (CCT) we obtained was tendentially lower than the mean value previously found for Yerkes captive chimpanzees. Close relationships, which were present in all female-female (FF) and some male-female (MF) dyads, positively affected reconciliation rates. When only adult PC-MC pairs (157) were considered, the mean TCTs and CCTs did not differ significantly. When we focused on types of PC affiliative contact, in the case of consolation we found a striking preference for sociosexual patterns. As to the relative occurrence of consolation and reconciliation, the highest level of the former was found in the absence of the latter. When reconciliation took place, consolation generally preceded it, suggesting that consolation may be a substitutive behavior. Our findings suggest that even if reconciliation remains the best option, consolation may be an alternative substitute for reconciliation that is used to buffer the tension originating from an unresolved conflict. Reconciliation and consolation are complex phenomena that are probably related to the life history of a group. Given that few studies have been conducted on this subject, we can not at this time make any generalizations regarding conflict resolution in certain species by comparing results among studies. PMID- 14752811 TI - Genotyping from semen of wild Japanese macaques (Macaca fuscata). AB - The noninvasive collection of animal cells is crucial for DNA analyses in wild populations that cannot be disturbed by capture. We describe the collection of 68 semen samples following copulation and masturbation events in wild habituated and nonhabituated troops of Japanese macaques on the protected island of Yakushima. We used this DNA to amplify 390 base pairs (bp) of the mitochondrial DNA control region in 16 individuals from eight troops, and found a monomorphic pattern in agreement with the low variability imposed by geographic isolation and female philopatry. We also amplified two microsatellite loci from samples collected after the resident males of a focal troop had copulated with different females. We found several different allele combinations in samples collected after the observed mating of a single male, indicating the presence of contaminant DNA, presumably from males that had previously mated with the same female. This discovery made it impossible to assign a given sample to a specific male except when the samples were recovered after masturbation events. Thus, it was not possible to test for kinship or estimate allele frequencies from the semen samples. The mixing of semen, and the pattern of sample collection observed in morphologically identified individuals support the notion that strong mating and sperm competition exists among resident and nonresident males. PMID- 14752812 TI - Behavioral study in the gray mouse lemur (Microcebus murinus) using compounds considered sweet by humans. AB - This study presents the results from two-bottle preference (TBP) tests performed on the gray mouse lemur (Microcebus murinus), a small Malagasy primate. We found that of 18 compounds considered sweet by humans, M. murinus preferred only six: D tryptophan, dulcin, fructose, sucrose, SC45647, and xylitol. The animals neither preferred nor rejected acesulfame-K, alitame, aspartame, N-4-cyanophenyl-N' cyanoguanidineacetate (CCGA), cyanosuosan, cyclamate, monellin, saccharin, suosan, super-aspartame, N-trifluoroacetyl-L-glutamyl-4-aminophenylcarbonitrile (TGC), and thaumatin. Together with previously recorded taste-nerve responses in M. murinus to acesulfame-K, alitame, aspartame, cyclamate, monellin, saccharin, and suosan [Hellekant et al., Chem Senses 18:307-320, 1993b], the current results suggest that these compounds either do not taste sweet to M. murinus or they have an aversive taste component. In this work we also relate these findings to phylogeny. PMID- 14752813 TI - The impact of Hepatology. PMID- 14752815 TI - Structural biology of hepatitis C virus. AB - Hepatitis C virus (HCV) causes acute and chronic liver disease in humans, including chronic hepatitis, cirrhosis, and hepatocellular carcinoma. Studies of this virus have been hampered by the lack of a productive cell culture system; most information thus has been obtained from analysis of the HCV genome, heterologous expression systems, in vitro and in vivo models, and structural analyses. Structural analyses of HCV components provide an essential framework for understanding of the molecular mechanisms of HCV polyprotein processing, RNA replication, and virion assembly and may contribute to a better understanding of the pathogenesis of hepatitis C. Moreover, these analyses should allow the identification of novel targets for antiviral intervention and development of new strategies to prevent and combat viral hepatitis. This article reviews the current knowledge of HCV structural biology. PMID- 14752816 TI - Federally funded research in hepatology. PMID- 14752817 TI - Non-HFE hemochromatosis. PMID- 14752818 TI - Noninvasive markers of esophageal varices: another round, not the last. PMID- 14752819 TI - The importance of knowing your identity: sources of confusion in stem cell biology. PMID- 14752820 TI - Unusual suspects in primary biliary cirrhosis. PMID- 14752821 TI - Viral features of lamivudine resistant hepatitis B genotypes A and D. AB - Viral differences among lamivudine resistant hepatitis B (HBV) genotypes have not been yet investigated. Therefore, we analyzed the characteristics of these viral strains in vivo. Forty-one patients carrying lamivudine resistant HBV were enrolled. Twenty-six patients (63%) carried resistant HBV genotype A (group A) and 15 patients (37%) carried resistant HBV genotype D (group D). The rate of reverse transcriptase 204I mutants was significantly higher in group D (67%) compared with group A (19%), whereas rt204V mutants (81% in group A vs 33% in group D; P =.006) and rt180M mutants (81% in group A vs 40% in group D, P =.015) prevailed in group A. The median time of shift from rt204I to rt204V mutants was significantly shorter in group A (4 months in group A, >12 months in group D, P <.001). Additional resistance associated mutations were detected exclusively in group D (P =.004). In a multivariate analysis, HBV genotype (P =.039) and pretreatment serum HBV DNA (P =.001) were independently associated with emerging rt204I or rt204V mutants, respectively. Serum HBV copy numbers after emergence of resistance were higher in group A (mean log(10) 6.99 copies/ml; range 3-9) compared with group D (mean log(10) 6.1 copies/ml; range 3.3-8; P =.04). There was no difference between both groups regarding core promoter/precore mutations, viral turnover, and number of flares or disease progression during follow-up. In conclusion, the mutational pattern during selection of lamivudine resistant HBV strains differs between genotypes A and D. This may have consequences for a salvage regimen initiated for treatment of lamivudine resistant HBV. PMID- 14752822 TI - High prevalence of occult hepatitis B in Baltimore injection drug users. AB - Occult hepatitis B is defined by the presence of hepatitis B virus (HBV) DNA in a serum or liver in the absence of hepatitis B surface antigen (HBsAg). The prevalence and clinical correlates of occult hepatitis B remain incompletely defined. A cross-sectional study was performed to determine the prevalence of occult hepatitis B in a high-risk cohort composed of 188 injection drug users in Baltimore, Maryland. All individuals had chronic hepatitis C viral infections confirmed by RNA detection and liver biopsy. Serologic assays for HBsAg and core antibody (HBcAb) were performed. Serum HBV DNA was detected using the COBAS HBV AMPLICOR monitor assay (lower limit of detection, 200 HBV copies per milliliter) and a semi-nested polymerase chain reaction (PCR) assay (lower limit of detection, 15 HBV copies per milliliter). Although almost all individuals (96%) were anti-HBC positive, only 8 of 188 (4%) were HBsAg positive. Occult hepatitis B was not identified using the COBAS assay, but was found in 81 of 180 (45%) of individuals using semi-nested PCR. Of the 8 HBsAg positive individuals, HBV DNA was found in 1/8 using the COBAS assay and 6/8 using the nested PCR assay. Overall, liver disease was mild, with a median serum alanine aminotransferase (ALT) of 38 IU/L, median activity grade of 3/18, and median fibrosis stage of 1/6. No association was found between the serum AST (aspartate aminotransferase), activity grade, or stage of liver disease and the presence of occult hepatitis B. Serum ALT levels were slightly higher in patients without occult hepatitis B (46 vs. 35 IU/L), and the median years since first injection drug use was somewhat longer in those without occult hepatitis B (24 vs. 20 years). In conclusion, although further research is needed to assess its clinical significance, there is a high prevalence of occult HBV infection in this cohort of HCV-infected injection drug users. PMID- 14752823 TI - Pediatric fulminant hepatic failure in endemic areas of hepatitis B infection: 15 years after universal hepatitis B vaccination. AB - To investigate the role of hepatitis B virus (HBV) infection in pediatric fulminant hepatic failure (FHF) after the launch of universal HBV vaccination, the authors analyzed the data from patients with FHF collected from a nationwide collaborative study group. Children aged 1 month to 15 years who were diagnosed with FHF (62 males and 33 females) between 1985-1999 were included. HBV infection (hepatitis B surface antigen [HBsAg] and/or immunoglobulin M hepatitis B core antibody [IgM anti-HBc] seropositive) accounted for 46% (43 of 95 cases) of all the cases of FHF. The average annual incidence of FHF in the time period 1985 1999 was 0.053/100,000 in the group of patients ages 1-15 years and 1.29/100,000 in those patients age < 1 year. Approximately 61% (58 of 95 cases) of all FHF cases were infants. The percentage of HBV infection was found to be higher in infants (57%) compared with children ages 1-15 years (27%) (P = 0.004). The incidence rate ratio of those patients age < 1 year to those ages 1-15 years was 54.2 for HBV-positive FHF and 15.2 for HBV-negative FHF. Maternal HBsAg was found to be positive in 97% of the infants with HBV-positive FHF, and hepatitis B e antigen (HBeAg) was found to be negative in 84% of these infants. Approximately 74% of all HBV-positive FHF patients and 81% of the infantile HBV-positive patients had been vaccinated. In conclusion, within the first 15 years of universal vaccination, HBV was found to rarely cause FHF in children age > 1 year but remained a significant cause of FHF in infants. HBV-positive FHF was prone to develop in infants born to HBeAg-negative, HBsAg-carrier mothers; these infants had not received hepatitis B immunoglobulin according to the vaccination program in place. PMID- 14752824 TI - Identification of HBV DNA sequences that are predictive of response to lamivudine therapy. AB - Numerous studies have shown that resistance to long-term lamivudine therapy occurs in as many as (2/3) of hepatitis B virus (HBV) chronic carriers. Additional studies have shown that reversion of HBV mutations in the precore/core promoter region conferring an HBeAg-negative phenotype/genotype can occur in up to 30% of lamivudine-treated patients. In this study, sequences of the HBV polymerase and precore/core coding regions in 26 HBV-infected patients (24 with HBeAg-negative virus infection, 25 genotype D, 1 genotype A) treated for 27 to 53 months with lamivudine were analyzed to determine the relationship between pretreatment HBV DNA sequence patterns and long-term treatment response, and the effect of therapy on the status of HBV precore mutations. Reversions of precore mutations A1762T/G1764A and G1896A were observed in 29% and 25% of patients, respectively, but none became HBeAg-positive. These data are consistent with previously published reversion frequencies for 2 other groups of lamivudine treated patients. Two naturally-occurring DNA polymorphisms at aa91 and aa256 of the HBV polymerase were observed in the pretreatment serum samples, which correlated with extended treatment failure. In conclusion, reversion of mutations conferring an HBeAg-negative phenotype occur relatively frequently under lamivudine therapy. Furthermore, at least in HBeAg-negative patients infected predominantly with HBV genotype D, specific viral DNA sequences which are present before therapy appear to be useful as predictors of long-term response to lamivudine treatment. PMID- 14752825 TI - Health-related quality of life in active injecting drug users with and without chronic hepatitis C virus infection. AB - This study assessed the effect of chronic hepatitis C virus (HCV) infection on the health-related quality of life (HRQOL) of injecting drug users, comparing the HRQOL of injecting drug users with and without chronic HCV infection. The study included 199 injecting drug users of more than 18 years of age who participated in a needle exchange program. Blood samples were tested for the presence of HCV RNA in serum with a polymerase chain reaction method. HRQOL was measured using the questionnaire SF-36, measuring HRQOL over the last 4 wk. The HCV RNA test was positive in 102 (51%) and negative in 97 (49%) subjects. The HRQOL scores of actively injecting drug users were markedly reduced compared to the population norm. However, we did not find poorer HRQOL in injecting drug users with chronic HCV infection than in injecting drug users without HCV infection. HCV RNA positive injecting drug users who were aware of the infection had lower HRQOL scores than those unaware of the infection in 4 of the 8 SF-36 dimensions (general health, physical functioning, physical role, and vitality). HCV RNA negative subjects, who believed they were infected, scored worse in one dimension (general health) compared to those who did not believe they were infected. In conclusion, chronic HCV infection per se did not negatively affect the HRQOL of active injecting drug users. Those who thought they were infected had a lower HRQOL scores than those who believed they were not infected. PMID- 14752826 TI - Reactive oxygen species suppress hepatitis C virus RNA replication in human hepatoma cells. AB - Hepatitis C virus (HCV) is a positive-stranded RNA virus that causes severe liver diseases, such as cirrhosis and hepatocellular carcinoma. HCV uses an RNA dependent RNA polymerase to replicate its genome and an internal ribosomal entry site to translate its proteins. HCV infection is characterized by an increase in the concentrations of reactive oxygen species (ROS), the effect of which on HCV replication has yet to be determined. In this report, we investigated the effect of ROS on HCV replication, using a bicistronic subgenomic RNA replicon and a genomic RNA that can replicate in human hepatoma cells. The treatment with peroxide at concentrations that did not deplete intracellular glutathione or induce cell death resulted in significant decreases in the HCV RNA level in the cells. This response could be partially reversed by the antioxidant N acetylcysteine. Further studies indicated that such a suppressive response to ROS was not due to the suppression of HCV protein synthesis or the destabilization of HCV RNA. Rather, it occurred rapidly at the level of RNA replication. ROS appeared to disrupt active HCV replication complexes, as they reduced the amount of NS3 and NS5A in the subcellular fraction where active HCV RNA replication complexes were found. In conclusion, our results show that ROS can rapidly inhibit HCV RNA replication in human hepatoma cells. The increased ROS levels in hepatitis C patients may therefore play an important role in the suppression of HCV replication. PMID- 14752827 TI - Long-term outcome (35 years) of hepatitis C after acquisition of infection through mini transfusions of blood given at birth. AB - Long-term follow up studies of hepatitis C virus (HCV) infection rarely exceed 20 25 yr. We studied the outcome of HCV infection in 35-yr-old adults infected at birth (1968) through mini transfusions of blood. A retrospective-prospective study was carried out. The cohort included 31 individuals who were given mini blood transfusions (21-30 ml) collected from a donor subsequently revealed to be HCV infected. At enrollment (1998), 18 of 31 (58.1%) recipients had anti-HCV antibody and 16 (88.9%) of them were HCV-RNA positive. All viremic recipients and the infectious donor had the same genotype 1b. Sequence analysis of E1/E2 and NS5b regions, coupled with phylogenetic analysis, indicated that HCV isolates from donor/recipients were linked. Eleven of the 16 viremic recipients gave consent to liver biopsy. Nine had no fibrosis or mild portal fibrosis and 2 had either discrete (Ishak's staging 3) or marked (Ishak's staging 4) fibrosis. During the prospective follow-up period (1998-2003), 2 patients were given therapy, one of whom achieved sustained clinical and virologic response. A second biopsy, performed in 5 patients at a 5 yr interval, revealed no substantial modifications in 4 cases and progression from absence of fibrosis to mild portal fibrosis in the fifth. In conclusion, taking into account the limited study sample, these findings suggest that HCV infection acquired early in life shows a slow progression and mild outcome during the first 35 yr of infection. PMID- 14752828 TI - Differential modulation of rat hepatic stellate phenotype by natural and synthetic retinoids. AB - Activation of hepatic stellate cells (HSC) is a central event in the pathogenesis of liver fibrosis during chronic liver injury. We examined the expression of retinoic acid (RAR) and retinoid X receptors (RXR) during HSC activation and evaluated the influence of natural and synthetic retinoic acids (RA) on the phenotype of culture-activated HSC. The expression of the major RAR/RXR subtypes and isoforms was analyzed by Northern hybridization. Presence of functional receptor proteins was established by gel shift analysis. Retinoic acids, RAR, and RXR selective agonists and an RAR antagonist were used to evaluate the effects of retinoid signalling on matrix synthesis by Northern blotting and immunoprecipitation, and on cell proliferation by BrdU incorporation. The 9-cisRA and synthetic RXR agonists reduced HSC proliferation and synthesis of collagen I and fibronectin. All-trans RA and RAR agonists both reduced the synthesis of collagen I, collagen III, and fibronectin, but showed a different effect on cell proliferation. Synthetic RAR agonists did not affect HSC proliferation, indicating that ATRA inhibits cell growth independent of its interaction with RARs. In contrast, RAR specific antagonists enhance HSC proliferation and demonstrate that RARs control proliferation in a negative way. In conclusion, natural RAs and synthetic RAR or RXR specific ligands exert differential effects on activated HSC. Our observations may explain prior divergent results obtained following retinoid administration to cultured stellate cells or to animals subjected to fibrogenic stimuli. PMID- 14752829 TI - AQP4 transfected into mouse cholangiocytes promotes water transport in biliary epithelia. AB - Rodent cholangiocytes express 6 of the 11 known channel proteins called aquaporins (AQPs) that are involved in transcellular water transport in mammals. However, clarifying the role of AQPs in mediating water transport in biliary epithelia has been limited in part because of the absence of physiologically relevant experimental models. In this study, we established a novel AQP4 transfected polarized mouse cholangiocyte cell line suitable for functional studies of transepithelial water transport, and, using this model, we define the importance of this AQP in water transport across biliary epithelia. Polarized normal mouse cholangiocytes (NMCs) lacking endogenous AQP4 were transfected stably with functional AQP4 or cotransfected with functional AQP4 and a transport deficient AQP4 dominant negative mutant using a retroviral delivery system. In transfected NMCs, AQP4 is expressed on both the mRNA and protein levels and is localized at both the apical and basolateral membranes. In nontransfected NMCs, the transcellular water flow, P(f), value was relatively high (i.e., 16.4 +/- 3.2 microm/sec) and likely was a reflection of endogenous expression of AQP1 and AQP8. In NMCs transfected with AQP4, P(f) increased to 75.7 +/- 1.4 microm/sec, that is, by 4.6-fold, indicating the contribution of AQP4 in channel-mediated water transport across MNCs monolayer. In cotransfected NMCs, AQP4 dominant negative reduced P(f) twofold; no changes in P(f) were observed in NMCs transfected with the empty vector. In conclusion, we developed a novel polarized mouse cholangiocyte monolayer model, allowing direct study of AQP4-mediated water transport by biliary epithelia and generated data providing additional support for the importance of AQP4 in cholangiocyte water transport. PMID- 14752830 TI - Spontaneous cholecysto- and hepatolithiasis in Mdr2-/- mice: a model for low phospholipid-associated cholelithiasis. AB - Previously, we identified needle-like and filamentous, putatively "anhydrous" cholesterol crystallization in vitro at very low phospholipid concentrations in model and native biles. Our aim now was to address whether spontaneous gallstone formation occurs in Mdr2 (Abcb4) knockout mice that are characterized by phospholipid-deficient bile. Biliary phenotypes and cholesterol crystallization sequences in fresh gallbladder biles and non-fixed liver sections were determined by direct and polarizing light microscopy. The physical chemical nature and composition of crystals and stones were determined by sucrose density centrifugation and before mass and infrared spectroscopy. Gallbladder biles of Mdr2(-/-) mice precipitate needle-like cholesterol crystals at 12 weeks of age on chow. After 15 weeks, more than 50% of Mdr2(-/-) mice develop gallbladder stones, with female mice displaying a markedly higher gallstone-susceptibility. Although gallbladder biles of Mdr2(-/-) mice contain only traces ( or =18 years, treated before age 20 years on Children's Cancer Group (CCG) ALL protocols, and 396 sibling controls completed a telephone interview and the Harter Adult Self-Perception Profile (ASPP). RESULTS: Survivors global self-worth scores were significantly lower than sibling controls (mean 3.09 vs. 3.18; P = 0.022). Unemployed survivors reported lower global self-worth scores than employed (mean 2.77 vs. 3.12; P = 0.0001), whereas employment status was not associated with self-worth in controls. Among survivors, predictors of negative self-concept included unemployment (odds ratio (OR) = 2.87; 95% CI: 1.50 5.50), and believing that cancer treatment limited employability (OR = 3.17; 95% CI: 1.79-5.62). Unemployment increased the odds for negative self-concept among survivors who received combinations of central nervous system (CNS) irradiation (CRT) and intrathecal methotrexate (IT-MTX), except high CRT with no or low dose IT-MTX. Employed survivors who perceived that treatment limited their employability showed increased odds of negative self-concept for all treatment groups compared to those who did not. Minority ethnic group membership was a borderline significant predictor of negative self-concept (OR = 1.79; 95% CI: 0.94-3.33). CONCLUSIONS: Global self-worth was significantly lower in ALL survivors than sibling controls, however, 81% of survivors had positive self concept. Survivor subgroups most vulnerable to negative self-concept were the unemployed survivors, believing that cancer treatment affected employability, and ethnic minority group members. Targeted intervention may have greater clinical relevance for these subgroups. PMID- 14752861 TI - Starting an adolescent cancer unit: why does it take so long? AB - A pragmatic approach is discussed based on the authors' differing experiences while designing new units specifically for adolescents with cancer in Leeds and Newcastle upon Tyne. Planning and implementation are complex and very time consuming. Areas to be considered include: formally identifying needs and adapting to existing local circumstances, convening a working group and involving potential stakeholders at an early stage, designing a suitable physical space, recruiting and integrating staff to create a supportive environment, obtaining financial support, and developing operational policies. For successful running of an adolescent unit, old prejudices must be disavowed and new models of medical care considered. PMID- 14752862 TI - Caring for a child with cancer--a systematic review. AB - OBJECTIVE: To report the results of a systematic review to determine the effects of parents' behavior on children's coping with cancer. METHODS: Searches were conducted using Medline, EMBASE, PsycLit, and articles were subsequently selected on the basis of predefined criteria. RESULTS: Twenty-four papers were identified. There were associations between parenting behaviors and child distress both before and during medical procedures. Parents who criticized the child, or apologized for what was happening had children who were more distressed. Parents who were very permissive had more problems with adherence to treatment regimens. CONCLUSIONS: Parents' behavior is critical in determining children's responses during procedures and adherence to home care. There has been less work concerning how parents manage more everyday problems, such as encouraging the child to go to school. Longitudinal studies are recommended to determine how parenting behaviors affect longer term child adjustment. These findings may be helpful for clinic staff to understand parents' reactions, and may also inform the content of intervention programs. PMID- 14752863 TI - Treatment of CNS malignant rhabdoid tumors. AB - BACKGROUND: Central Nervous System (CNS) rhabdoid tumours are a highly malignant group of neoplasms usually occurring in children under 2 years of age with characteristic histopathologic findings but unclear histiogenesis and almost uniformly fatal outcome. There is still no proven curative therapy available. PROCEDURE: The clinical course and the successful outcome of therapy in two children with primary CNS rhabdoid tumour are described in this context. Both children had subtotal excision of the primary tumour and received chemotherapy based on the SIOP Malignant Mesenchymal Tumour (MMT-95) protocol with addition of triple intrathecal chemotherapy. Following this, one of the patients received high dose therapy (busulphan and thiotepa), whereas the other had craniospinal radiotherapy with a boost to the primary site. RESULTS: The treatment was reasonably well tolerated and both patients are alive with no evidence of disease 52 months and 65 months after the primary diagnosis. Their favourable outcomes are compared with those of 49 others reported in the literature. CONCLUSIONS: Intensified therapy (with autologous bone marrow transplantation and intrathecal chemotherapy) may improve the prognosis of patients with malignant rhabdoid tumour. PMID- 14752864 TI - High-dose chemotherapy with autologous stem-cell rescue in the treatment of patients with recurrent non-cerebellar primitive neuroectodermal tumors. AB - BACKGROUND: Recurrent non-cerebellar primitive neuroectodermal tumors (PNETs) carry a dismal prognosis when treated with conventional chemotherapy alone. XSWe tested the efficacy of high-dose chemotherapy (HDC) followed by autologous stem cell rescue (ASCR) in this setting. PROCEDURE: Eligibility mandated either minimal residual disease or evidence of chemosensitivity before HDC. Conditioning consisted of carboplatin (CBDCA) (500 mg/m(2) or AUC = 7 mg/ml min using the Calvert formula) on days -8 to -6, thiotepa (300 mg/m(2)), and etoposide (250 mg/m(2)) on days -5 to -3. Irradiation was given post HDC selectively. RESULTS: Among 17 patients treated in this study, there were eight pineoblastoma(s) (pineo), seven cortical PNETs, and two arising elsewhere. Relapse was either local (nine) or metastatic to the brain (four) or spine (four). Two patients received HDC as the sole therapy for recurrence; additionally, eight underwent surgical debulking before HDC, and nine received irradiation, including six after HDC. Median age at ASCR was 3.9 years. Two patients died of toxicity (11%) and ten experienced tumor relapse (range: 23-361 days post ASCR). Five patients with cortical PNETs remain alive disease-free (median follow-up: 8.3 years); four of them received irradiation post HDC. The difference in 5-year event-free survival (EFS) between patients with pineo and other supratentorial PNETs was significant (0 vs. 62.5 +/- 17%, P = 0.0065). Both surgery at relapse and irradiation post HDC were favorable prognostic factors (P = 0.006 and 0.01, respectively). CONCLUSIONS: Patients with recurrent cortical PNETs can be cured with this strategy. Surgical debulking before, and irradiation after HDC play an important role in treatment success. PMID- 14752865 TI - Biodistribution of post-therapeutic versus diagnostic (131)I-MIBG scans in children with neuroblastoma. AB - BACKGROUND: To evaluate the biodistribution of therapeutic (131)I metaiodobenzylguanidine (MIBG) and assess the sensitivity of diagnostic versus therapeutic (131)I-MIBG scans to detect metastatic disease. PROCEDURE: This retrospective study included 44 diagnostic and post-therapy scans (PTS) in 18 children with neuroblastoma treated with (131)I-MIBG (2.0-33.1 GBq). The findings of diagnostic scans (DS) (2.6-44.4 MBq) were compared to those of corresponding PTS. RESULTS: In terms of biodistribution, the PTS identified (131)I-MIBG activity in one or more patients in the following regions not detected on the DS: nasal mucosa, cerebellum, central brain, adrenals, spleen, kidneys, thyroid, salivary glands, lower halves of the lungs, bladder, bowel, and an incisional scar. Conversely, the DS identified activity in the thorax, heart, kidneys, and bladder each in one patient without being visualized on the PTS. In terms of sensitivity to detect metastatic disease, 210 lesions were seen on the PTS compared to 151 on the DS. The PTS demonstrated sites of disease not evident in the DS in 16 cases. CONCLUSIONS: The biodistribution of (131)I-MIBG is different using therapeutic doses as compared to pre-therapy doses. (131)I-MIBG imaging following high therapeutic doses often reveals sites of occult metastatic disease that may be clinically relevant. PMID- 14752866 TI - Clonal proliferation of T-Cell large granular lymphocytes. AB - BACKGROUND: T-cell large granular lymphocyte (T-LGL) leukemia is a disorder only rarely reported in children. We diagnosed a new case of a clonal T-LGL proliferation in a 6-year-old girl presenting with severe neutropenia and pure red blood cell aplasia (PRCA). PROCEDURE: Flow cytometric analysis including TCR Vbeta repertoire analysis and molecular studies using reverse transcriptase polymerase chain reaction (RT-PCR), PCR heteroduplex analysis and GeneScan analysis were performed to investigate the clonal nature of the T-LGL. RESULTS: Flow cytometric analysis revealed a Vbeta3 clonal nature. Molecular studies identified a clonal Vbeta3-Cbeta RT-PCR product. Both PCR heteroduplex analysis and GeneScan analysis found clonal TCRB and TCRG gene rearrangements. CONCLUSIONS: An underlying T-LGL leukemia should be investigated in the diagnostic evaluation of acquired PRCA and neutropenia in young children. Both flow cytometric and molecular analyses can be used to establish the clonal nature of T-LGL. PMID- 14752867 TI - Association of cytogenetic abnormalities with detection of BCR-ABL fusion transcripts in children with T-lineage lymphoproliferative diseases (T-ALL and T NHL). AB - Detection of Philadelphia chromosome (Ph) in childhood T-lineage acute lymphoproliferative disorders is a rare event. Additional cytogenetic abnormalities are particularly uncommon in ALL. We here report two cases with T lineage acute lymphoproliferative disorders (T-ALL and T-NHL) presenting with both cytogenetic alterations and BCR-ABL fusion transcripts, associated with an aggressive presentation and a poor outcome. We point out firstly on the cytogenetic aberrations, supporting the hypothesis of multi-lineage involvement of ALL expressing Ph chromosome; secondly, on the persistence of T-cell leukemic clone detected by minimal residual disease (MRD) analysis, despite of the early disappearance of BCR-ABL fusion transcript. PMID- 14752868 TI - A very uncommon presentation of bilateral Wilms tumor. PMID- 14752869 TI - Cefepime vs. Meropenem as empirical therapy for neutropenic fever in children with lymphoma and solid tumours. PMID- 14752870 TI - Treatment of vincristine-induced bilateral ptosis with pyridoxine and pyridostigmine. PMID- 14752871 TI - Secondary malignant neoplasms of the bladder after cyclophosphamide treatment for childhood acute lymphocytic leukemia. PMID- 14752872 TI - Extraovarian primary peritoneal carcinoma in a child. PMID- 14752873 TI - Leukemic cells and the cytokine patchwork. AB - BACKGROUND: In leukemia, the clonal population is characterized by a hierarchical organization. Although the majority of the leukemic population is generated after post-determinic divisions, a subset of cells retain undifferentiated "blast" morphology. In addition, leukemic cells often have numerical or structural chromosomal abnormalities, aberrant gene expression patterns, and abnormal cell surface marker profiles. Despite these differences when compared to normal bone marrow and blood cells, leukemic cell survival and proliferation, just like that of normal progenitor cells, is influenced by hematopoietic growth factors. A major issue is whether differential regulation of normal and leukemic hematopoietic cells by cytokines can be exploited in antileukemic treatment or, in contrast, whether in vivo cytokine therapy may even be harmful to the patients. PROCEDURE: Here we review the results of recent experimental and clinical observations that investigated the influence of cytokines on leukemic cell growth and differentiation in vitro and in vivo. RESULTS: The majority of studies indicate that hematopoietic growth factors are involved in the regulation of proliferation and terminal differentiation of leukemic blast cells. Genetic aberrations involving cytokines or their receptors may contribute to leukemogenesis. Abundant interactions, cross-lineage stimulation, and aberrant response patterns seem to transform the complex cytokine network regulation of normal hematopoiesis into an even more interlaced "patchwork" that controls leukemic hematopoiesis. CONCLUSIONS: Since hematopoietic growth factors are present in high serum concentrations in patients with acute leukemia and myelodysplastic syndromes, consequences of possible interactions should be kept in mind even when well-defined human recombinant factors in single application are to be involved in antileukemic protocols. PMID- 14752874 TI - Increased risk for aplastic anemia and myelodysplastic syndrome in individuals lacking glutathione S-transferase genes. AB - BACKGROUND: Aplastic anemia (AA) and myelodysplastic syndrome (MDS) are marrow failure states that may be associated with chromosomal instability. An absence of the glutathione S-transferase (GST) enzyme may genetically predispose individuals to AA or MDS. PROCEDURE AND RESULTS: To test this hypothesis, we determined the GSTM1 and GSTT1 genotypes in a total of 196 patients using multiplex PCR. The GSTT1 null genotype was found to be overrepresented in Caucasian, Asian, and Hispanic patients with either AA or MDS. We confirmed a difference in the expected frequency of the GSTM1 null genotype in Caucasian MDS patients. The double null GSTM1/GSTT1 genotype was also overrepresented in Caucasian AA and MDS patients. In our population, 26% of AA patients and 40% of MDS patients had a chromosomal abnormality identified by karyotype or FISH analyses for chromosomes 7 and 8. Patients with AA and the GSTT1 null genotype had an increased frequency of chromosomal abnormalities (P = 0.003). CONCLUSION: There seems to be an increased risk for AA and MDS in individuals lacking GSTT1 or both GSTM1/GSTT1. PMID- 14752875 TI - Effects of physical therapy intervention for children with acute lymphoblastic leukemia. AB - BACKGROUND: The purpose of this study was to examine the effects of physical therapy intervention in children with acute lymphoblastic leukemia (ALL). PROCEDURE: Twenty-eight children aged 4-15 years were randomly assigned to an intervention or control group. The intervention group received five sessions of physical therapy and was instructed to perform an individualized home exercise program consisting of ankle dorsiflexion stretching, lower extremity strengthening, and aerobic exercise. RESULTS: After 4 months children who received physical therapy intervention had significantly improved ankle dorsiflexion active range of motion and knee extension strength (P < 0.01). Differences were not found between the two groups for any of the other dependent variables. CONCLUSIONS: Physical therapy intervention for children with ALL receiving maintenance chemotherapy improved two body functions important for normal gait. Physical therapy programs initiated earlier in treatment and with greater emphasis on endurance activities may also improve stamina and quality of life (QOL). PMID- 14752876 TI - Is alveolar histotype a prognostic factor in paratesticular rhabdomyosarcoma? The experience of Italian and German Soft Tissue Sarcoma Cooperative Group. AB - PURPOSE: To ascertain whether alveolar histology retains its adverse prognostic role in the subset of paratesticular rhabdomyosarcoma (RMS) patients, generally characterized by a very good outcome. PATIENTS AND METHODS: Twenty pediatric patients were treated over a 25-year period using the protocols of the Italian and German Soft Tissue Sarcoma Cooperative Groups. Clinical characteristics at presentation were much the same as in non-alveolar patients. RESULTS: The proportion of patients with alveolar histotype (8%) in paratesticular site was lower than in the general RMS population (20-30%). With a median follow-up of 122 months, 5-year EFS and OS were 78 and 89%, respectively. CONCLUSION: Our data suggest a distinctly better clinical behavior of paratesticular alveolar RMS than when the disease occurs at other sites. These patients were more intensively treated than the embryonal cases, however, so-although a treatment intensity reduction may be desiderable-the idea of eliminating the alkylating agents (as in low-risk embryonal paratesticular RMS) must be considered with great caution. PMID- 14752877 TI - Outcome of pediatric recurrent and refractory malignant solid tumors following ifosfamide/carboplatin/etoposide (ICE): A phase II study in a pediatric oncology centre in Brazil. AB - BACKGROUND: Therapeutic trials using ifosfamide,carboplatin, and etoposide (ICE) regimen have suggested many patients with refractory/relapsed solid tumors may be rendered free of disease using this combination. The aim of this study was to assess the feasibility, response and toxicity of this regimen in a Brazilian Pediatric Oncology Centre. PROCEDURE: ICE, consisting of ifosfamide 3 g/m(2)/day x3 plus Mesna, etoposide 160 mg/m(2)/day x3, and carboplatin 400 mg/m(2)/day x2 was given at 21-28 days intervals. G-CSF was used for patients with hematological recovery greater than 4 weeks. RESULTS: Twenty-one patients younger than 18 years of age treated with this regimen were studied between July 1996 and November 2000. A total of 93 courses of ICE were administered. Leucopenia/neutropenia grades 3/4 were the most frequent severe adverse effects (82%). Grade 3/4 thrombocytopenia occurred in 73% of cycles, and anemia was common (61%). The most common grade 3/4 non-hematologic toxicities were fever and proved infection occurring in 25% and 14% of the courses, respectively. Two patients developed renal tubular damage. Other less common grade 3/4 non-hematologic toxicities included nausea and vomiting, hepatotoxicity, and stomatitis. The overall response rate (complete response/partial response) was 53%. The median progression-free interval was 15.3 months, with a median survival of 24.2 months. CONCLUSIONS: Although ICE was associated with severe myelosuppression it produced objective response in about half of the cases. The most important non hematological toxicity was severe renal tubular damage. This regimen should only be used whenever hematological and infectious supportive care is available. PMID- 14752878 TI - Significance of pleural effusion at diagnosis of Wilms tumor. AB - BACKGROUND: Pleural effusion is uncommon at diagnosis of Wilms tumor. Because the clinical significance of this finding has not been reported, a retrospective review of this entity was conducted. PROCEDURE: The radiology reports and medical records of 233 patients with Wilms tumor who were treated at St. Jude Children's Research Hospital between 1985 and 2001 were reviewed. RESULTS: Ten of the 233 patients (4.3%) had pleural effusions detected on pre-nephrectomy imaging studies. Computed tomography (CT) scans were more sensitive than chest X-rays in identifying pleural effusions. All of the patients with pleural effusions had tumors of favorable histology. Two patients had stage II disease, five had stage III disease, two had stage IV disease, and one had stage V (bilateral) disease. All of the effusions occurred on the same side as the primary kidney tumors and six effusions were associated with ascites. The two effusions that were tapped were negative for tumor cells. All 10 patients in our series were alive without recurrence with a median follow-up duration of 68 months, yet only two patients were treated with lung irradiation. CONCLUSION: Pleural effusion at presentation of Wilms tumor is uncommon and not associated with adverse prognosis. We recommend that pleural effusions be tapped whenever feasible if the results will change therapy. Patients with pleural effusions that are small and difficult to tap may be treated according to their local stage without pulmonary irradiation. Further analysis of pleural effusions in a larger group of patients is necessary to confirm our preliminary observations. PMID- 14752879 TI - Carboplatin-epirubicin regimen for the treatment of hepatoblastoma. AB - BACKGROUND: In order to lower the long-term toxicity of chemotherapy for hepatoblastoma patients, a prospective study was designed based on pre-operative chemotherapy combining carboplatin and epirubicin (CE). PROCEDURES: Patients under 16 years of age with an epithelial hepatic tumor diagnosed by ultrasound or CT scan and a high serum alpha-foetoprotein (AFP) level were eligible. Patients were treated with a pre-operative chemotherapy regimen combining carboplatin 600 mg/m(2) and epirubicin 80 mg/m(2). Tumor resectability was assessed after four courses given at 3-week intervals. After surgery, patients were given two more courses of CE. Response was assessed based on a drop in serum AFP and tumor shrinkage. RESULTS: Between July 1988 and August 1995, 27 patients with a hepatoblastoma were included. The initial PRETEXT group according to the SIOPEL classification was: group 2 (5 pts), group 3 (15 pts), group 4 (5 pts), and 2 pts were not assessed. Six patients had lung metastases. Response was partial response (PR) in 20/27 (74%) patients, disease was stable in 3 and 4 had progressive disease (PD). A complete surgical resection was performed in 21 pts. Five-year overall and disease-free survival (DFS) were respectively 56% (95%CI: 37-72%) and 63% (95%CI: 44-78%). During the same time period, 7 pts with a hepatocellular carcinoma were treated according to this protocol. Only one achieved a PR. Toxicity was mostly hematologic with > or =grade 3 leukopenia in 23% of the courses, > or =grade 3 thrombocytopenia in 29% of the courses and anemia in 22%. CONCLUSION: The CE protocol is feasible and efficient in hepatoblastoma. However, only a randomized study will permit a valid comparison of the efficacy of cisplatin and carboplatin for the treatment of these patients. PMID- 14752880 TI - Medulloblastoma in childhood: Impact of radiation technique upon the outcome of treatment. AB - BACKGROUND: Medulloblastoma is an infratentorial primitive neuroectodermal tumour, diagnosed in paediatric population. The radiotherapy is an essential method of treatment for these tumours. The impact of technical quality of radiation therapy on survival was recently considered. In this study treatment related variables are analysed with a special focus on radiotherapy technical factors. PROCEDURE: The population of 158 children with a diagnosis of medulloblastoma had been undergoing postoperative radiotherapy at MCMCC in Warsaw in the period 1983-1997. The medical data of these patients were analysed. Simulation films of the whole-brain irradiation fields were retrospectively reviewed at 112 patients. The distance from the field margin to the cribriform plate and to the floor of the temporal fossa was assessed and correlated with supratentorial relapse. RESULTS: Five-year overall survival (OS) and 5-year disease free survival (DFS) were 46% and 40%, respectively. In multivariate analysis gender (P = 0.008), neurological status before radiotherapy (P = 0.029), M-stage (P < 0.001) and sequence of craniospinal irradiation (P = 0.019) were significant prognostic factors for OS. For DFS significant factors were M-stage (P < 0.001) and neurological status (P < 0.001). The cranial fields were not fully correctly covered at 43% patients. The field incorrectness was correlated with isolated supratentorial failure (P = 0.049). CONCLUSIONS: Our results are similar to those published in literature. M0-stage was the most powerful favourable prognostic factor. Male gender and neurological status before radiation treatment were associated with poor survival. Also protracted radiotherapy and quality of radiation technique may have an impact on the outcome. PMID- 14752881 TI - Long-term follow-up of survivors of childhood cancer in the UK. AB - BACKGROUND: Childhood cancer is rare, but there are now good survival prospects and in the UK approximately 1 in 1,000 young adults is a survivor of childhood cancer. There are many adverse health outcomes associated with the treatment of childhood cancer often arising several years after completion of treatment. The aim of this study was to quantify the long-term clinical follow-up practices concerning survivors of childhood cancer. PROCEDURE: A cross-sectional postal survey of 22 treatment centres of the United Kingdom Children's Cancer Study Group (UKCCSG) clinicians was carried out as well as a cross-sectional postal survey of general practitioners of most adult survivors of childhood cancer in Britain. RESULTS: Subsequent to 5 years after the end of treatment: 52% of UKCCSG clinicians follow-up all survivors for life, while 45% discharge some patients. Of those clinicians discharging: over 50% discharged benign, stage I or tumors treated with surgery alone, in contrast 16% reported discharging all or most patients; almost all (97%) clinicians discharged to a general practitioner. Only 14% of clinicians reported nurses undertook a specialist role. Sixty-five percent of the 10,979 general practitioners reported that their patient was not on regular hospital follow-up. CONCLUSIONS: There are wide variations in the extent to which survivors of childhood cancer are discharged from hospital follow-up. There is a need for regularly updated national guidelines concerning the levels of follow-up required for specific groups of survivors defined principally by the treatment they received. PMID- 14752882 TI - Epidemiologic analysis of 1,442 children and adolescents registered in the German germ cell tumor protocols. AB - BACKGROUND: Germ cell tumors (GCTs) constitute a heterogeneous group of tumors that significantly vary with respect to their clinical presentation and biology. The objective of this analysis was to analyze a large population-based pediatric cohort of GCTs and to evaluate the parameters age, sex, site of the tumor, histology, and potential correlations between these parameters. PROCEDURE: Between 1981 and 2000, 1,442 patients were prospectively enrolled onto the German protocols for testicular and non-testicular GCTs. Tumors were histologically classified according to the WHO. RESULTS: We observed a bimodal age distribution with a first peak during infancy and a second after the onset of puberty. At birth, almost all tumors were teratomas, sometimes with microfoci of yolk sac tumor, which on the other hand, was the predominant histology during childhood. After the onset of puberty, germinomatous GCTs represented the most frequent histological subtype, and malignant non-germinomatous GCTs often presented as mixed tumors with choriocarcinoma and embryonal carcinoma components. During infancy, non-gonadal GCTs accounted for the majority of GCTs, while after the onset of puberty, gonadal GCTs predominated. Notably, among non-gonadal GCTs, there was a female predominance during childhood and a strong male predominance during adolescence. CONCLUSIONS: Two separate groups of GCTs with distinct clinical features relevant for differential diagnosis and the diagnostic assessment can be distinguished. This observation correlates with genetic studies that reveal different genetic changes in childhood and adolescence GCTs. Further studies are needed to elucidate the molecular mechanisms of germ cell and GCT development that account for the age- and sex-dependent clinical manifestation. PMID- 14752883 TI - Differentiated thyroid carcinoma in childhood and adolescence-clinical course and role of radioiodine. AB - BACKGROUND: Differentiated thyroid carcinoma (DTC) in childhood has a good prognosis despite a high incidence of relapse. The use of radioactive iodine (RAI) has not been well established. PROCEDURE: This is a review of 60 patients less than 21 years of age; mean follow-up was 14 years. RESULTS: Patients had a higher relapse rate with papillary thyroid carcinoma (PTC) than with follicular thyroid carcinoma (FTC): 24.5 vs. 9.1%. Compared with 997 patients with age > or = 21, patients <21 years of age had a higher female to male ratio (7.6 vs. 3.9), higher incidence of nodal metastasis (45 vs. 28%), and lung metastasis (15 vs. 7.8%), and improved 10-year cause-specific survival (CSS) (98.3 vs. 89.5%). The 10-year rates of CSS, local-regional failure-free survival (LRFFS), and distant metastasis failure-free survival (DMFFS) for the young patients were 98.3, 79.3, and 90.7%, respectively. In patients with no distant metastasis at presentation, RAI improved 10-year LRFFS (71.9 vs. 86.5%; P = 0.04). At last follow-up, 10 of 12 patients (80%) with local-regional (LR) relapse and five of nine patients (55.6%) with distant metastasis were rendered disease-free. No patient has experienced a second malignancy. CONCLUSIONS: Prognosis of DTC in young patients was good. Patients with LR relapse and distant metastasis had a high rate of remission after treatment. RAI treatment can reduce the rate of LR relapse in patients with no distant metastasis and result in complete remission in half of those with distant metastasis. No patient experienced a second malignancy. PMID- 14752884 TI - Gist one of those things? PMID- 14752885 TI - Gastric stromal tumors in children. AB - Gastrointestinal stromal tumors (GIST) in children are rare and their behavior has been regarded as difficult to predict on pathological criteria. We report our experience with two gastric GISTs in children aged 10 and 11 years. Both remain alive and free of disease at 5 years and 2 years respectively. Comparison of the pathological features in the resected specimens with a recently proposed guidelines for predicting outcome in this group of tumors is reported. PMID- 14752886 TI - Malignant infantile osteopetrosis and primary pulmonary hypertension: a new combination? AB - Malignant infantile osteopetrosis (MIOP), a rare genetic disorder of the osteoclast, is fatal without hematopoietic stem cell transplantation. Primary pulmonary hypertension (PPH), a rare progressive disorder of the pulmonary circulation, is predominantly fatal in the absence of successful therapy. A clinical association between these two disorders has not been recognized and a pathophysiologic link between osteoclast function and pulmonary vascular pressure as a rationale for such an association is not readily apparent. Here, we report five infants with MIOP, without cardiac abnormalities, who were found to have PPH after undergoing stem cell transplantation. We suggest that PPH may be linked to a specific variant of MIOP and recognizing the potential for pulmonary hypertension in children with MIOP may lead to a more rapid diagnosis and life saving intervention. PMID- 14752887 TI - Ex vivo drug resistance in childhood acute myeloid leukemia on relapse is not higher than at first diagnosis. AB - Relapsed pediatric patients with acute myeloid leukemia (AML) have a poor clinical prognosis. The aim of this study was the analysis of the ex vivo drug resistance profile on relapse in childhood AML in comparison to newly diagnosed AML. The results of 98 pediatric AML samples tested by the MTT assay were analyzed. Eighteen samples (18%) were excluded from the further analysis due to spontaneous apoptosis of blasts in 4-days culture, low percentage of myeloblasts in the sample either in the beginning or at the end of the assay, infection, or formation of clots in the sample. Finally, ex vivo drug resistance of 20 relapsed samples were compared with that of 60 de novo AML, including 9 matched pairs. Up to 18 drugs were tested for each patient. No significant differences between drug resistance at diagnosis and at relapse in AML was found, neither for the whole groups of patients, nor for matched pairs only. Possibly, relatively good sensitivity of myeloblasts on relapse was found against melphalan, thiotepa, 4 HOO-ifosfamide, and cladribine. In summary, cellular drug resistance in childhood AML at relapse is not higher than at first diagnosis. These observations suggest that other, than cellular drug resistance, factors play a key role in therapy failure of relapsed childhood AML. PMID- 14752888 TI - Anemia and a large abdominal tumor in an adolescent. PMID- 14752889 TI - Re: Is this glaucoma? PMID- 14752890 TI - NIH report card. PMID- 14752891 TI - Prognostic value of myeloperoxidase in patients with chest pain. PMID- 14752892 TI - Hospital use and survival among Veterans Affairs beneficiaries. PMID- 14752893 TI - HIV/AIDS among African Americans and US women: minority and young women. AB - The HIV/AIDS pandemic continues to be a major health crisis facing the African American community. Although African Americans make up only about twelve percent of the U.S. population, they accounted for half of the new HIV infections reported in the United States in 2001. Numerous studies suggest that many new infections occur among young African Americans. In the early 1980s, HIV/AIDS was considered mostly a gay white male disease in the United States. Today, the pandemic has expanded and the disease is also a major health problem in the African American community, where men and women of every age and sexual orientation are affected. PMID- 14752894 TI - The nursing shortage. PMID- 14752895 TI - Update: HIV prevention: a medical perspective for persons living with HIV. PMID- 14752896 TI - Tobacco use: an addiction. PMID- 14752897 TI - [Work conditions and occupational morbidity among agricultural workers]. AB - Comparative evaluation of work conditions, risk factors and health state of agricultural workers could serve as forecasting criteria of occupational morbidity level. PMID- 14752898 TI - [Work experience at the Saratov Regional Center for Sanitary and Epidemiologic Supervision in occupational hygiene for agricultural workers]. AB - The authors studied contemporary peculiarities of work conditions of individuals engaged into agriculture, cause-effect relationships in occupational morbidity and priority directions in occupational diseases prophylaxis. PMID- 14752899 TI - [Pathophysiologic aspects of development of occupational diseases and their laboratory diagnosis (review of the literature)]. PMID- 14752900 TI - [Occupational hygiene during pesticide use]. AB - The study covered assortment and amount of pesticides varying in purpose and chemical class and applied in Saratov region in 2002 vs in 1980s and 1990s. Findings are changed assortment, occurrence of new generation pesticides, significantly lower pesticide load, and seed mordants predominant over other pesticide groups. These findings necessitate special attention to occupational hygiene for health preservation among rural agricultural workers. PMID- 14752901 TI - [Evaluation of workplaces concerning work conditions and its role in occupational risk management among agricultural workers]. AB - Evaluation of workplaces with subsequent certification on work safety could serve as a means solving problem of lower occupational risk for health and life among agricultural workers. PMID- 14752902 TI - [Monitoring of myocardial contraction function during an annual occupational cycle in agricultural machine operators]. AB - Infrasound cardiography enables to diagnose premorbid conditions, determine myocardial contractility, regulate work intensity and increase efficiency of prophylactic measures. PMID- 14752903 TI - [Epidemiologic features of occupational brucellosis in Saratov region]. AB - Retrospective epidemiologic analysis proved general and occupational morbidity with brucellosis to decrease in Saratov region. This evidence could be due to better episootic situation for the infection. The authors justified necessity of complex measures on acute and chronic brucellosis prevention. PMID- 14752904 TI - [Methodical aspects of health of individuals working in social and hygienic monitoring system]. PMID- 14752905 TI - [Hygienic and ergonomic features of work of workers engaged in milk-processing enterprises]. PMID- 14752906 TI - [Main factors of occupational risk among mobile agricultural machinery operators]. PMID- 14752907 TI - [Basic hygiene of using insecticides on private farms]. PMID- 14752908 TI - [HF-treatment in rehabilitation of individuals subjected to ecologic hazards]. PMID- 14752909 TI - [Hygienic problems of optimizing rural water supply in the land along the Volga]. PMID- 14752910 TI - [Databases for planning hygiene research]. PMID- 14752911 TI - Abnormal aneurysmal dilatation of the internal carotid artery on angiography without abnormal finding at surgery: a case report. AB - After trauma, the next most common cause of subarachnoid hemorrhage (SAH) is aneurysm rupture. Aneurysms can be of any size and shape and most commonly are found at the anterior or posterior communicating cerebral arteries. Although magnetic resonance angiography is improving in diagnostic accuracy, routine catheter angiography is still considered the "gold standard" for diagnosis. We report a case in which an abnormal dilatation of the supraclinoid internal carotid artery was diagnosed as an aneurysm following catheter angiography; at surgery this area appeared normal. PMID- 14752912 TI - Primary adrenal lymphoma presenting as adrenal insufficiency. A case report and review of literature. AB - Primary adrenal lymphoma (PAL) with adrenal insufficiency is a rare entity that has a unique presentation and prognosis when compared to other high-grade B-cell Non-Hodgkin's Lymphomas (NHL). Radiologic characteristics and image-guided biopsy are helpful in diagnosis. Current chemotherapy of choice is CHOP [Cyclophosphamide, Hydroxydaunomycin (doxorubicin), Oncovin (vincristine sulfate) and Prednisone]. More cases need to be documented to formulate an effective approach to PAL. PMID- 14752913 TI - Emerging Pseudomonas aeruginosa resistance: implications in clinical practice. PMID- 14752914 TI - Bioterrorism preparedness--Part II. Smallpox vaccination in a hospital setting. AB - The threat of using smallpox as an agent for bioterrorism resulted in a directive for the creation of smallpox response teams. In Connecticut, The Commissioner of the Department of Public Health convened public health and hospital leadership to plan for the vaccination of these teams. The purpose of this paper is to provide a description of the vaccination program at Hartford Hospital, a Center of Excellence for Bioterrorism Preparedness, and to report the results of a survey of the vaccinees regarding the vaccination experience. Ninety persons were vaccinated. Six individuals experienced low-grade fever and 10 had axillary node swelling. One individual experienced significant fatigue. A total of six persons lost time from work. Four lost one day and two persons lost between four to five days of work. There was no autoinoculation, transfer inoculation, vaccinia or any other significant complication. Survey results indicate that most vaccinees felt positive about the experience. PMID- 14752915 TI - An assessment of Hispanic health status. AB - The health-care systems in urban communities are facing major challenges. They are being forced to reduce costs while maintaining and improving quality. Compounding these systems issues are the inequalities in health access and barriers that persist within certain segments of the population. Hispanic Americans make up a significant and growing segment of American society; one particularly important issue for Hispanics is staying healthy. Lack of health insurance, environmental stressors, limited access to provider services, and language barrier are a few of the social inequalities that add to the difficulty in receiving health care. To reduce the inequalities and improve access to quality health care, an assessment may be necessary. An investigation into the trends in the health status, beliefs, and practices of Hispanics would be appropriate in order to address their health needs. These inquiries must go beyond having knowledge of different countries of familial origin to include knowledge of differing cultures, socioeconomic status, and experiences within the health-care system. PMID- 14752916 TI - [Role of glycoprotein IIb/IIIa inhibitors in the management of acute coronary syndromes]. AB - Antiplatelet therapy is a cornerstone in the medical management of acute coronary syndromes. Three classes of antiplatelet drugs are available in this setting: acetylsalicylic acid, thienopyridines, and glycoprotein IIb/IIIa antagonists. During the last ten years, numerous clinical trials have been conducted in large populations of patients suffering from acute coronary syndromes. Further investigations are in progress. In the light of these results, the respective roles of the different antiplatelet drugs have been more precisely defined, in terms of class preference as well as in terms of the combination of several antiplatelet drugs and of antiplatelet drugs with other therapies, including non fractionated--or low molecular weight--heparin and non-invasive or invasive revascularisation procedures. In the present article, we review the results of the major published or non published trials that addressed the role of glycoprotein IIb/IIIa antagonists in the management of acute coronary syndromes. Based on these results, the current therapeutic guidelines for clinical practice issued by the American and European cardiology societies are given in the conclusion, with their level of evidence. PMID- 14752917 TI - [Cardiac resynchronization in congestive heart failure: role of Doppler echocardiography in patient selection]. AB - Cardiac echodoppler is an efficient tool to analyse cardiac mechanic loop. Diastolic and systolic desynchronisation criteria are analysed. It should allow a more efficient selection of patients who could take benefit of resynchronization therapy, but also to choose resynchronization system, to program optimal atrioventricular delay, interventricular delay and to evaluate resynchronization efficiency. However, this tool should be evaluated in large studies in order to compare it to electrocardiogram. PMID- 14752918 TI - [Value and limitations of carotid sinus massage in healthy elderly individuals. Evaluation of diagnostic criteria for hypersensitive carotid sinus syndrome]. AB - OBJECTIVE: To study the value and limitations of carotid sinus massage in healthy individuals older then 50 years of age, in order to assess the validity of the widely used criteria for the diagnosis of hypersensitive carotid sinus syndrome. METHODS: Right and left carotid sinus massage was performed in 120 healthy individuals older then 50 years of age (74 males and 46 females aged 59 +/- 7 years) who had no prior history of syncope, pre-syncope, or abnormal bradyarrhythmia, during EKG monitoring. RESULTS: Hypersensitivity of the carotid sinus (HSCS) defined as a pause > 3 s, was found in 28 individuals (23.3%). The pause was induced by right carotid sinus massage in 20 cases and by left carotid sinus massage in 8 cases. Patients demonstrating HSCS were older compared to those who did not develop a pause. One or multiple blocked P wave was seen in 16 cases when carotid sinus massage was performed on the left side compared to 8 with right carotid sinus massage. CONCLUSION: In healthy individuals older than 50 years of age, it is not rare to induce a significant pause following carotid sinus massage. This limits the validity of the widely used criteria for the diagnosis of hypersensitive carotid sinus syndrome. Additional criteria are proposed to make this diagnosis in front of a brief loss of consciousness. PMID- 14752919 TI - [Sphingolipid signaling: a potential pathway for TNF-alpha induced preconditioning]. AB - Although cytokine activation has long been recognized to associate with cardiac ischemia and reperfusion, the concept that these cytokines may enhance some cardioprotective mechanisms has only recently been considered. Ischemic preconditioning is a biologic phenomenon that activates innate cytoprotective programs in the heart. Ischemic preconditioning has been described where a transient non-lethal ischemic "trigger" or endogenous molecules produced/released by ischemia enables the tissue to become more resistant/tolerant to subsequent potentially lethal ischemia. The mechanisms and signalling events involved in this cytoprotective program still remain obscure. Recently, it has been suggested that cytokine activation including tumour necrosis factor (TNF alpha) may play a key role in the preconditioned phenotype. Moreover, new studies have given the evidence that the exploration of cytokine-activated sphingolipid signalling pathways may enhance our understanding of the preconditioning program. PMID- 14752920 TI - [Primary cardiac sarcoma: report of 3 cases and review of the literature]. AB - Primary cardiac sarcoma is a rare tumor with a poor prognosis. We report 3 cases with a review of literature about this disease. There were 2 males and 1 woman. The main symptoms were thoracic pain. The clinical features were various and the thoracic ultra sonography exam allowed the diagnosis in the 3 cases. All patients had surgical remove of their cardiac tumor followed by chemotherapy. All of them died within 13 to 36 months after the diagnosis. Primary cardiac sarcoma has a poor prognosis with a mean survival less than 12 months. PMID- 14752921 TI - [A strange case of phlebitis]. AB - Report of a rare case concerning an elderly man (81 years) suffering from a leiomyosarcoma of the leg already metastasized to the lungs, discovered in connection with a suspected thrombophlebitis. This sarcoma developed from the smooth muscle of a leg vessel, probably a vein. Leiomyosarcoma is a malignant mesenchymal tumor of specialized connective tissue, with a strong potential for local proliferation and metastatic spread. It usually involves the uterine muscle or the wall of the digestive tract, as well as the large vessels of the abdomen and thorax, the prostate very seldom, and only exceptionally a peripheral vein as in this case. The diagnosis suggested by imaging techniques (in particular MRI) is first and foremost immunohistochemical. The treatment is surgical when possible, associated with radiotherapy and chemotherapy as appropriate. The prognosis is especially poor when the diagnosis is made at the metastatic stage. PMID- 14752922 TI - [Ventricular fibrillation induction by transesophageal stimulation in a patient with valvular heart disease]. AB - Rapid atrial transesophageal stimulation is currently used to stop atrial flutter or tachycardia. We report a case complicated of ventricular fibrillation. It is possible to pace the ventricles by transesophageal stimulation in 2-10% of cases. The ventricular stimulation might be dangerous in patients with heart disease. Therefore, transesophageal stimulation must be performed in specialized environment. PMID- 14752923 TI - [Postpartum cardiomyopathy revealed by acute lower limb ischemia]. AB - Peripartum cardiomyopathy is an uncommon disease defined as a dilated cardiomyopathy during puerperium, with left ventricular dysfunction (ejection fraction < 45%) without any other etiology. The etiology of this disease remains uncertain and it can be revealed in a variety of ways. Thrombo-embolic complications may be, although infrequently, the initial manifestation of peripartum cardiomyopathy, which is usually an intracardiac thrombosis. Lower extremity embolism is uncommon. The case reported is about a 39-year-old woman, multiparous, who presented, 40 days after delivery, a global heart failure with atrial fibrillation, revealed by left lower extremity thromboembolism. After echocardiographic and etiologic examinations, the diagnosis was established as peripartum cardiomyopathy. It evolved favourably after 2 months of medical treatment: the symptoms and cardiomegaly decreased, left ventricular systolic function was improved. PMID- 14752924 TI - [Tuberous sclerosis and cardiac tumor (case report)]. AB - We report the case of a cardiac tumor, which is found in a systematic cardiac examination of an 11 year old girl who had a tuberous sclerosis. The interest of this observation is to show the importance of a general examination, especially cardiac, within this pathology. Tuberous sclerosis is an hereditary disease, associated with rhabdomyomas in 50% of cases or more. Cardiac rhabdomyomas are frequently multiple and detected in utero in some cases. Conversely, 60% of these tumors are seen in a context of tuberous sclerosis. Rhabdomyomas are the most common cardiac tumors of infants and children, the large majority occurring in patients younger than one year and are diagnosed more and more in foetal echocardiography. PMID- 14752925 TI - [Pulmonary scintigraphy in the diagnosis of pulmonary embolism]. PMID- 14752927 TI - [Coronaviruses. Causes of SARS and other infections]. PMID- 14752928 TI - [Causes, symptoms and therapeutic methods for glaucoma]. PMID- 14752929 TI - [Hyperthyroidism in type 1 diabetics]. PMID- 14752930 TI - [Administration times for intranasal drops]. PMID- 14752931 TI - [Lymphoproliferative disorders caused by hereditary genetic defects]. PMID- 14752932 TI - [Recent advances in the diagnosis of primary immunodeficiency]. PMID- 14752933 TI - [Investigation of pathophysiology of Graves' disease by animal models]. PMID- 14752934 TI - [A case of Sjogren's syndrome associated with chronic hepatitis C and sarcoidosis]. AB - A 68-year-old female had been treated for chronic type C hepatitis at our department since June, 1992. In August of the same year, swelling of the right eyelid developed and she was diagnosed as having sarcoidosis on the basis of uveitis, skin lesions, and bilateral hilar lymphadenopathy (BHL) on chest X-ray. With steroid therapy, the symptom improved and BHL disappeared. In July, 2001, dryness of the mouth and dry eyes developed, and she was diagnosed as having Sjogren's syndrome (SjS). This case may be precious in discussing the pathophysiology of sarcoidosis and SjS including the association with hepatitis C virus infection. PMID- 14752935 TI - [A case of systemic lupus erythematosus complicated with psoriasis vulgaris]. AB - A 49-years-old female admitted to our hospital because of skin eruptions on the extremities in 1985. She had suffered from polyarthralgia, skin eruptions since 1983. Physical examinations revealed discoid lesion, central nervous system involvement, and polyarthritis. Laboratory tests revealed leukopenia, thrombocytopenia, and hypocomplementemia. Antinuclear antibody, ant-DNA antibody, LE test were positive. From these findings, she was diagnosed as systemic lupus erythematosus (SLE). She developed lupus peritonitis in 1990 and 1994, which was successfully treated by steroid pulse therapy. Since then, the activity of SLE was in good control under administration of prednisolone 10 mg/day. Chilblain lupus was seen from 1993, Raynaud's phenomenon from 1996, and she further developed subcutaneous induration on her chest, back and upper extremities in 1999. Skin biopsy findings were compatible with lupus panniculitis. In 2002, erythematous patches with scales were observed on her right hand and left knee, and these skin lesions were histologically diagnosed as psoriasis vulgaris. An autoimmune response similar to SLE is speculated in psoriasis. We describe a rare case of SLE with various skin lesions including psoriasis vulgaris. PMID- 14752936 TI - Detection of anti-SS-A/Ro and anti-SS-B/La antibodies of IgA and IgG isotypes in saliva and sera of patients with Sjogren's syndrome. AB - To assess the frequency and the possibility of local production of autoantibodies against SS-A/Ro and SS-B/La in patients with primary Sjogren's syndrome (SS), serum and saliva samples were obtained from 42 patients with SS, 10 with rheumatoid arthritis without sicca syndrome, and 12 healthy volunteers. Autoantibodies were detected using enzyme-linked immunosorbent assay and immunoblotting. The frequencies of IgA anti-SS-A antibody, IgA anti-SS-B antibody, IgG anti-SS-A antibody and IgG anti-SS-B antibody in serum from SS patients were 45%, 50%, 43% and 21%, respectively. The frequencies of IgA anti-SS A antibody, IgA anti-SS-B antibody, IgG anti-SS-A antibody and IgG anti-SS-B antibody in saliva from SS patients were 31%, 33%, 40%, and 19%, respectively. We also found secretory IgA anti-SS-A and anti-SS-B antibodies accompanying secretory components in saliva and sera in representative SS patients. Significant correlations were found between serum and salivary levels of IgA anti SS-A antibodies, and between serum and salivary levels of IgA anti-SS-B antibodies in SS patients. Significant correlations were also found between serum and salivary levels of IgG anti-SS-A antibodies, and between serum and salivary levels of IgG anti-SS-B antibodies in SS patients. Immunoblot analysis confirmed the presence of IgA-class autoantibodies against SS-A and SS-B in saliva and serum from representative patients. The presence of IgA- and IgG-class autoantibodies against SS-A and SS-B and those accompanying secretory components in saliva from SS patients suggests the local production of these antibodies and the relationship between local and systemic antibody responses. PMID- 14752937 TI - [Surgical science in future]. PMID- 14752938 TI - [A case of isolated hepatic malignant lymphoma representing characteristic findings on radiological imaging]. AB - We present an 80-year-old-man who showed characteristic radiological findings of secondary hepatic malignant lymphoma. The patient was first admitted to our hospital 6 months before with left cervical mass, which was diagnosed as malignant lymphoma by biopsy. After complete response to chemotherapy, a large hepatic mass appeared, as the serum value of lactic dehydrogenase elevated. CT/MRI showed a well-demarcated hypovascular mass, which was penetrated by the right hepatic vein. The apparent diffusion coefficient of the mass was 0.70 x 10( 3) mm2/s. The patient was diagnosed as relapse of malignant lymphoma, mainly based on these radiological findings reflecting the mass with high cellularity which grows without destruction of existing vascular structure, and additional chemotherapy was given. The hepatic mass disappeared almost all and the serum value of lactic dehydrogenase decreased with in normal range. The patient was discharged in a good condition 2 months later. PMID- 14752939 TI - [How do medical facilities cope with a new medical fee system? A case study of the Kumamoto medical area]. AB - In April 2000, a new medical fee system for hospitals designated for specific functions was introduced into the remuneration of medical services. A basic concept of this system is to pay a supplementary fee for hospital treatment during the acute treatment. In this study, in order to clarify the effect of this new system on hospital management, we performed the case study of three hospitals (Kumamoto National Hospital, Saiseikai Kumamoto Hospital and Kumamoto Chuo Hospital) located in Kumamoto medical area. As a result, in this area, the basic qualitative aspects of medical care environment, such as the number of hospital beds and long-term facilities, was found to be well equipped. In addition, three hospitals differ in following points: 1) characteristics of clinical department, 2) management strategy. In conclusion, the case of the Kumamoto medical area was considered to provide an important model of collaboration and co-operation between medical facilities toward the new medical fee system. PMID- 14752940 TI - Mindful meals: a holistic approach to eating. PMID- 14752941 TI - National Center for Complementary and Alternative Medicine: a valuable resource for all of us. PMID- 14752942 TI - Healthy fats: getting to the fat of the matter. PMID- 14752943 TI - What does it mean to grow older? PMID- 14752944 TI - Hear ye hear ye: a comprehensive ear care program for residents residing in long term care facilities. PMID- 14752945 TI - Severe acute respiratory syndrome (SARS) in the elderly. PMID- 14752946 TI - Reflections on working with families in long-term care during the SARS restrictions. PMID- 14752947 TI - A dialogue on existential-phenomenological thought in psychology and in nursing. Interview by William K. Cody. PMID- 14752948 TI - Review of research related to Watson's theory of caring. AB - Forty retrievable studies are reviewed, covering the period of 1988 to 2003, that were based on Watson's theory of caring. The author provides an overview of the studies, identifies measurement instruments developed, critiques the body of work, and offers suggestions for future research. PMID- 14752949 TI - Exploring ethical implications for acting faithfully in professional relationships. AB - The obligation to act faithfully poses ethical issues as nurses live nursing from day to day. In this column, the ethics of acting faithfully is explored in relation to four scenarios drawn from different realms of professional nursing. The scenarios illustrate ethical issues that may arise when attempting to uphold personal integrity and fulfill one's duty to act faithfully. The theoretical context of this discussion is the human becoming school of thought. PMID- 14752950 TI - A human becoming teaching-learning model. AB - Teaching-learning is a never-ending journey of giving-receiving in coming-to know. The purposes of this column are to distinguish among the traditional and human becoming paradigms of teaching-learning, and to set forth the essences, paradoxes, and processes of a human becoming teaching-learning model. PMID- 14752951 TI - Witnessing change with aspiring nurses: a human becoming teaching-learning process in nursing education. AB - Nurse educators have the opportunity to encourage meaningful reflections of nursing students. Dr. Rosemarie Rizzo Parse's teaching-learning processes provide a framework for such experiences. Student reflection through journaling and student participation in dialogue using these processes brings about an opportunity for students to discover new meaning for themselves and others. The process of how two nurse educators incorporated the human becoming teaching learning model into students' experiences is discussed. Excerpts of student journals, themes of student work, and considerations for future development of the teaching-learning model with students are discussed. PMID- 14752952 TI - Dementia, personhood, and nursing: learning from a nursing situation. AB - Appreciation of the personhood of people with dementia calls for care that looks beyond the disease to the person within. In this column the author discusses the concept of personhood for people with dementia and presents a method of teaching person-centered care developed within the theoretical framework of nursing as caring. The study of an aesthetic expression of a nursing situation, written by a nurse who cares for people with Alzheimer's disease in a nursing home, is presented to assist nurses to learn to see beyond the disease to the person and to develop relationships that nurture personhood. PMID- 14752953 TI - Conceptual models of nursing: international in scope and substance? The case of the Neuman systems model. Interview by Jacqueline Fawcett. PMID- 14752954 TI - Nursing: whose discipline is it anyway? AB - Nursing is variously described as a profession, a discipline, an occupation. The meanings we assign to such words and the expectations, demands, and responsibilities that each reveals to and/or exacts from those of us privileged to call ourselves registered nurse provides a splendid arena for viewing the struggle of nursing as an intellectual endeavor embedded in its own distinctive knowledge base, experiences, purposes, and values. Currently, pressure exerted within and without nursing to adopt the self-limiting potential and subordinated position that a professional discipline orientation and applied degree education confer on nursing is mounting. This article examines the relationships among historical events, contemporary influences, and confounding definitions that are integral to how we choose to guide and shape the evolution of nursing scholarship, practice, and education. The authors contend that the full significance of these relationships must be appreciated if nursing is to advance as a scholarly academic discipline and practice profession. PMID- 14752955 TI - The lived experience of feeling cared for: a human becoming perspective. AB - The purpose of this study was to answer the research question, What is the structure of the lived experience of feeling cared for? The participants were 10 women volunteers who were struggling with lack of economic, social, or interpersonal resources and who were or had been homeless. The Parse research method, a phenomenological-hermeneutic method, was used to discover the meaning of feeling cared for. The major finding of this study is the structure: Feeling cared for is contentment with intimate affiliations arising with salutary endeavors, while honoring uniqueness amid adversity. The structure provides knowledge about feeling cared for and its connection to health and quality of life. Feeling cared for is discussed in relation to the principles and concepts of human becoming and in relation to how it can inform nursing practice and future research. PMID- 14752956 TI - Health and well-being in early adolescents using Rogers' science of unitary human beings. AB - The purpose of this study was to examine the relationship of perceived field motion and human field rhythms to perceived health status, health conception, and well-being in early adolescents to determine which health-related variables are most compatible with Rogers' science of unitary human beings. The sample of 142 early adolescents responded to the study instruments in classrooms. The correlations between perceived field motion and each of the other variables were statistically significant, and increased in magnitude from perceived health status to health conception to well-being. These results give credence to Rogers' evolving philosophical belief that the term well-being is more compatible than health in her science. PMID- 14752957 TI - Toward a middle-range theory of adaptation to chronic pain. AB - Merton postulated that to significantly advance disciplinary knowledge, scholarship must include the development of middle-range theories that derive hypotheses, can be empirically investigated, and are linked to extant theories. The purpose of this paper is to present the strategies used to develop a middle range nursing theory of adaptation to chronic pain based on Merton's description. Analysis and synthesis of the theoretical and research literature provided the foundational, theory-building strategies used to develop the adaptation to chronic pain model. Theoretical substruction was then used to deduce the adaptation to chronic pain model from Roy's adaptation model. PMID- 14752958 TI - Understanding the importance of values and moral attitudes in nursing care in preserving human dignity. AB - The purpose of this investigation was to explore when nursing becomes an art, and to develop a theory/model of nursing as art. The overall design of the investigation was qualitative, and phenomenological-hermeneutic. The five substantial categories in the theory/model are: Invitation and confirmation, encounter, values, moral attitude and demeanor, the act of good will, and aesthetic communication. One main finding of this nursing investigation based on the caring perspective was that values and a thoroughly moral attitude and demeanor are central when nursing becomes an art. PMID- 14752959 TI - Throwing darts. PMID- 14752961 TI - Japanese markets for dental implants. PMID- 14752960 TI - European markets for dental implants. PMID- 14752962 TI - The use of implantoplasty and guided bone regeneration in the treatment of peri implantitis: two case reports. AB - A variety of treatment modalities have been proposed for the management of peri implantitis. These are mostly based on empiric experience and use the systemic administration of an antibiotic in conjunction with surgical intervention. To ensure decontamination of the affected implant surface(s), chemical and/or mechanical debridement is used. For textured implant surfaces, detoxification using implantoplasty could also give favorable results when used as part of the procedure. Two cases are reported in which implants developed localized peri implantitis lesions. Implantoplasty followed by topical tetracycline decontamination was used in conjunction with guided bone regeneration. In both cases, the procedures were effective in arresting disease and regenerating lost bone. These results suggest that the technique holds promise and should be investigated further. PMID- 14752963 TI - Extractions, implant placement, and immediate loading of mandibular implants: a case report of a functional fixed prosthesis in 5 hours. AB - The purpose of this report is to present a reliable surgical and prosthodontic protocol for immediate loading of implants, allowing patients to receive a fixed implant-supported prosthesis in a matter of hours. The surgical, prosthodontic, and laboratory steps for this protocol of immediate functionally loading the implants are described for the treatment of a 51-year-old woman who presented with a hopeless mandibular dentition and an edentulous maxillary arch. It is possible to decrease the treatment time in successfully restoring the patient's oral function by means of immediate functional loading of dental implants and immediately placing a fixed implant-supported prosthesis. Compared with the traditional implant protocols, this protocol of a one-visit approach for patient treatment 1) decreases the number of office visits; 2) decreases the treatment time; 3) reduces the patients' costs; 4) allows the patient to avoid wearing a removable interim prosthesis; and 5) increases the patients' acceptance of treatment while maintaining predictability in treating mandibular edentulism. PMID- 14752964 TI - Bone graft augmentation and dental implant treatment in a patient with osteogenesis imperfecta: review of the literature with a case report. AB - This case report with a review of the literature describes a patient with osteogenesis imperfecta who was treated with dental implants. To our knowledge, only one case report has been published in the English dental literature describing implant treatment in the osteogenesis imperfecta patient. The uncertainty in treatment confronted by all clinicians is the density of bone, which is essential for implant survival. PMID- 14752965 TI - Simplifying single-stage solid abutments: Part 2. Temporization. AB - Fabrication of a temporary crown can consume much chairside time when restoring a single-unit implant. A technique for rapid fabrication of a temporary crown for the ITI implant system is addressed so as to minimize time and provide an accurate-fitting esthetic interim crown. PMID- 14752966 TI - Combination syndrome: treatment with dental implants. AB - Occlusal plane problems are often not evaluated adequately. They can be left untreated or improperly treated. This article reviews one such problem known as Combination Syndrome. The treatment method described involves using a fixed mandibular prosthesis over implants that have been placed immediately after dental extractions. PMID- 14752967 TI - Dental implant design and its relationship to long-term implant success. AB - The purpose of this review is to evaluate the effects of the biomechanical aspects of dental implant design on the quality and strength of osseointegration, the bone-implant interface, and their relationships to the long-term success of dental implants. The engineering design of implants is based on many interrelated factors, including the geometry of the implant, mechanical properties, and the initial and long-term stability of the implant-tissue interface. There is no one "optimal" design criterion. However, implants can be engineered to maximize strength, interfacial stability, and load transfer by using different materials, surfaces, and thread designs. Limited information is currently available in addressing how implant thread design influences the overall implant success. Therefore, this article reviews and discusses design elements of various dental implant systems currently in use as they affect the quality of osseointegration and their relationship to overall long-term success patterns. PMID- 14752968 TI - Cortical bone regeneration with a synthetic cell-binding peptide: a histologic and histomorphometric pilot study. AB - PepGen P-15 is a combination natural anorganic bovine-derived hydroxyapatite matrix (ABM) coupled with a synthetic cell-binding peptide (P-15). This material has been reported to enhance bone formation in periodontal osseous defects. The aim of this study was to assess the effect of ABM/P-15 on the healing of cortical bone defects in rabbits. Five New Zealand rabbits were used. Two 8-mm bone defects were created in each tibia. Eight defects were filled with PepGen P-15, 8 defects with PepGen P-15 Flow, and 4 defects were used as a control group. A total of 20 defects were created. All rabbits were killed at 4 weeks. Block sections containing the defects were retrieved and the specimens processed for light microscopy examination. Newly formed bone was present in both test groups, whereas, in the control-group, only a scarce quantity of newly formed bone was present and the cortical defects had not been filled by the regenerated bone. Statistical evaluation showed that there were statistically significant differences between control sites and sites treated with P-15 and P-15 Flow (P = 0.0001), and also between sites treated with P-15 and P-15 Flow (P = 0.0001), respectively. No acute inflammatory infiltrate cells were visible in both of these groups. Both PepGen P-15 and PepGen P-15 Flow enhanced new bone formation in the cortical drilled defects, whereas control defects showed very little newly formed bone. PMID- 14752969 TI - Histologic evaluation of human biopsies after dental augmentation with a demineralized bone matrix putty. AB - To solve some of the problems inherent in bone regeneration, various types of graft materials, matrix, putty, and gel delivery systems have been developed. These deliver demineralized bone matrix (DBM) to a graft site and maintain it in an appropriate position to achieve favorable results. This prospective study reviewed 10 cases of extraction immediate grafting with a putty-DBM delivery system. Five patients were male and 5 female. At intervals ranging from 4 to 21 months postextraction grafting, bone cores were harvested at the time of surgical insertion of Replace endosteal two-stage implants. All patients were restored with single-tooth self-standing prostheses. Bone quality and quantity in each of the bone cores were evaluated. All 10 cases were completed with favorable outcomes. PMID- 14752970 TI - The influence of diabetes mellitus and insulin therapy on biomechanical retention around dental implants: a study in rabbits. AB - The oral rehabilitation by dental implants in patients with diabetes remains a controversial issue. The aim of this study was to evaluate the influence of diabetes mellitus and insulin therapy on the bone healing around dental implants using torque removal. Twenty-seven rabbits were randomly divided into 3 groups with 9 animals each: control (C) group, induced diabetic (D) group, and insulin treated diabetic (ITD) group (10 U/day). After 1 week, one implant was inserted at the tibial metaphysis of the animals. The glucose levels were periodically evaluated through the glucose-oxidase enzymatic method. The animals were killed at 4, 8, and 12 weeks after surgery and the biomechanical test was performed using a torque manometer. Statistically significant differences regarding the removal torque of the implant could not be found at 4 weeks (P = 0.2) among groups. Group C showed statistically higher values than groups D and ITD at the experimental periods of 8 (P = 0.0001 and P = 0.0002, respectively) and 12 weeks (P = 0.0053 and P = 0.001, respectively). There were no statistical differences between D and ITD groups in any of the experimental periods. Diabetes mellitus has negatively influenced the mechanical retention of implants placed at the tibial metaphysis of rabbits. Therapy with insulin did not induce any changes. PMID- 14752971 TI - Estrogen deficiency affects bone healing around titanium implants: a histometric study in rats. AB - The purpose of this study was to evaluate the influence of an estrogen-deficient state on bone around titanium implants placed in rats. Thirty female Wistar rats were divided into 2 groups: test (n = 15), ovariectomized rats (OVX); and control (n = 15), sham-operated rats. Screw-type titanium implants were placed bilaterally in rats 21 days after ovariectomy or sham surgery. After 60 days, the animals were killed and undecalcified sections obtained. Blood samples were collected to obtain serum levels of alkaline phosphatase at the time of killing. Bone-to-implant contact (BIC), bone area (BA) around the implants, and bone density (BD) in a 500 microns-wide zone lateral to the implant were obtained and arranged separately for the cortical (zone A) and cancellous (zone B) regions. In zone A, there was no significant difference between test and control groups regarding BIC and BD (P > 0.05). A lower BA was observed in the estrogen deficient animals (P < 0.05). In zone B, data analysis showed that estrogen deficiency could result in a lower percentage of BIC, BA, and BD (P < 0.05). In addition, a higher concentration of alkaline phosphatase was observed for the test group. An estrogen-deficient state could affect bone healing and bone density around titanium implants placed in rats, especially in the cancellous bone area. PMID- 14752972 TI - Declining physician workforce is crisis for state. PMID- 14752973 TI - Mississippi's physician labor force: current status and future challenges. AB - The literature review indicates that changes in Medicaid/Medicare reimbursement, large numbers of uninsured patients, the legal climate, and largely rural and chronically ill populations create a challenging environment for physicians practicing in Mississippi. As a largely rural state, many Mississippians find medical care to be physically distant, with most care being concentrated in a couple areas of the state. Given these factors, the legal climate in Mississippi and the top relocation decision factors, Mississippi will be further challenged in recruiting and retaining the numbers of general practitioners and specialists necessary to provide care to the state's population. The challenges that physicians are facing have led to challenges for health policy makers, in that physicians are difficult to recruit to Mississippi and, once here, difficult to retain as practitioners throughout their career. Four datasets were used in conjunction to analyze the demographic characteristics of Mississippi's physicians, including the age structure disaggregated by several other variables. Ultimately, the results were extended to impacts of recruitment, relocations, and retirement decisions of physicians who participated in the MSMDS. Briefly, demographic results indicate that Mississippi has a largely white physician population serving a nearly 40% minority population in Mississippi. The under representation of women within the medical profession in Mississippi means that women in the state might find it unusually challenging to find a female physician, particularly in rural areas where access to physicians is more limited in the first place. Mississippi has a high concentration of African-American patients with a low African-American physician presence. The proportion of physicians who are female is on the rise nationwide and within Mississippi, largely due to increasing enrollments of women in medical schools. Though variations exist within the groups of physicians identified as generalists, Mississippi is only slightly more likely than the nation to have specialists, rather than generalists (see Table Seven). Age structure analysis indicates that Delta physicians are older than physicians elsewhere in the state, that urban physicians are younger than rural physicians, and that our physician labor force is more highly concentrated between the ages of 35 and 54 than in the nation as a whole. Analyses concerning the future of the physician labor force indicate that a near majority of Mississippi's practicing physicians received their MD degree at UMC, but younger physicians are more likely to have been educated out-of-state than older physicians. Those who received their degrees elsewhere and chose to practice in Mississippi are more likely to be specialists (60%) than generalists (40%). Those physicians practicing in the state who were educated in-state are nearly equally as likely to be generalists (47%) as they are to be specialists (53%). Additionally, those approaching retirement are more likely to be generalists, yet the state is recruiting more generalists from recent medical school classes than in the past. Variations in intentions to recruit, relocate, and retire exist. However, most of the substantively important variation is across age groups and time in practice. There is little relevance of specialty or location within the state when examining variation in recruitment, relocation or retirement plans. Given the findings, policy research recommendations focus on improving the retention of UMC's graduates for practice in the state, improving retention of active physicians, increasing the recruitment of physicians from out of state, and easing difficulties associated with working part-time as a step toward retirement. With these changes in policy, it is possible that Mississippi can thwart a physician workforce shortage; however, without changes, with more physicians relocating, retiring early, or opting out of practicing in the state, the extant physician shortage will become more severe. Furthermore, without the data collection efforts mentioned here, there will be no means to assess whether policy changes are actually impacting the physician labor force. PMID- 14752974 TI - Pockets of wealth. Although revenue is tight for long-term care facilities, other sources exist--you just need to know where to look. Roundtable discussion. PMID- 14752975 TI - Moving 'ahead'. New report confirms there are many bright sides to senior housing. PMID- 14752976 TI - A distinctive outlook. Industry financial benchmarks support 'asset quality' in assisted living. PMID- 14752977 TI - One-stop shopping. Cultivating a good relationship with your distributor pays big dividends. PMID- 14752978 TI - 'Home-grown' caregivers. An in-house CNA program can help you train and retain quality staff. PMID- 14752979 TI - Changing psychographics. Residents are demanding more as their perception of value changes. PMID- 14752980 TI - Get the money! Excuses you can expect when your debtors don't pay their invoices on time--and what you can do about them. PMID- 14752981 TI - Discover your writer within through ONS publishing opportunities. PMID- 14752982 TI - Actual data concerning the brain--immune system interface. AB - After a brief presentation of the immune system as sensorial and effector organ, which recognizes and defends against cellular aggressions, the main psycho-neuro endocrine components of immune reaction regulation and modulation will be shown. Both central nervous structures that control the hormonal emissions, the vegetative innervation of the lymphoid organs as well as the afferent neurohumoral pathways involved in the making of the self-regulating and neuromodulating circuits of the humoral and cellular immune responses will be mentioned. An important position will be held by the interrelations between the hypothalamus-pituitary-corticoadrenal gland, the sympathetic-parasympathetic efferent pathways and the chemical messengers (hormones, neurotransmitters, interleukins, neurotrophins) which make possible the bi-directional neuroimmune communication for maintaining the homeostatic balances on this third effector pathway, too. Also will be presented experimental proof concerning the ability of central neurons to secrete neuromodulator cytokines and the presence of specific receptors for the various neuroactive molecules within lymphoid organs and circulating lymphocytes. To close, the psychoemotional components of the neuro immunomodulator circuits will be mentioned, using as examples the changes induced by stress generally and oxidative stress in particular. PMID- 14752983 TI - Photodynamic therapy and some clinical applications in oncology. AB - Photodynamic therapy (PDT), a new treatment modality for localized cancers involving the selective interaction of visible light with photosensitizers, such as hematoporphyrin derivatives (HpD) or dihematoporphyrin ether/ester (DHE) (Photofrin II). Photodynamic therapy of malignant tumors includes biological, photochemical and photophysical processes. These processes involve: (i) absorption of photosensitizing agent; (ii) selective retention of photosensitizer in tumors and (iii) irradiation of sensitized tumor by laser irradiation. This paper provides a review of photosensitizers, photochemistry, subcellular targets, side effects and lasers involved in photodynamic therapy. In addition, gradual increase in knowledge related to in vivo and in vitro mechanisms of action of PDT, as well as some clinical applications of photodynamic therapy are presented. PMID- 14752984 TI - The level of interleukin-2 and of soluble interleukin-2 receptor in patients with Balkan nephropathy. AB - Balkan Nephropathy (BN) is defined as a clinical entity with unknown etiology. The involvement of immune system in pathogenesis of BN is not well defined yet. The aim of this study was to gain more insight into the cellular immune mechanisms in BN. We determined some factors implied in cellular immunity, such as the serum level of IL-2 and of IL-2 soluble receptor (sIL-2R), and the presence of IL-2 receptor alpha chain (CD25) on T cells membrane. The study was performed on 15 patients with BN, 15 patients with Chronic Pyelonephritis (CPN), and 10 healthy controls from a non-endemic area. Our study showed no significant differences between IL-2 level and CD25+ cells percentage in CPN compared to controls, but a significantly increased level of sIL-2R. The BN sIL-2R is significantly lower than sIL-2R in CPN, and associates an important T cell activation (high CD25+ presence, elevated IL-2 level) compared to CPN. Our conclusion is that while the high sIL-2R level could down modulate T cell activity in CPN, BN sIL-2R level is ineffective in limiting the activation effects of IL-2 on T cells. The results suggest that cellular immunity could have a role in the pathogenesis of N. PMID- 14752985 TI - Molecular cloning of a functional cis-acting, Bam HI-flanked, 1.6 Kb 'Mob' cassette for demonstrating rapid conversion of col EI origin-DNA cloning vectors into conjugal form. AB - A 1.6 Kb mobilization (Mob) fragment originating from broad host range IncP plasmid RP4 is effectively cloned into two different Col EI-origin based cloning vectors, pBluescript II SK+ and pT-Adv, to generate pPAR-I and pPAR-II, respectively. The vectors have different genetic markers and were demonstrated to get mobilized at significant frequency into a laboratory and an enteroroxigenic strain of Escherichia coli with all the genetic markers of the recombinant clones expressing efficiently in the recipient host cells. Important restriction endonuclease recognition sequences in the multiple cloning sites of the conjugal vector DNA molecules remained unique. Significance and relevance of the current study with regard to other gene delivery system in gram negative bacteria are discussed. PMID- 14752986 TI - Quo vadis bioterrorism research? PMID- 14752987 TI - [Obesity epidemic has hit Europe]. PMID- 14752988 TI - [Economic and health: history about trans fatty acids]. PMID- 14752989 TI - [Denmark is the first county in the world to forbid the use of industrially produced fatty acids]. PMID- 14752990 TI - [Obesity in Europe]. PMID- 14752991 TI - [D-vitamin: old paradoxes and new perspectives]. PMID- 14752992 TI - [Incidence of primary hyperparathyroidism, frequency of surgical interventions and mortality based on the data from the National Patient Registry]. PMID- 14752993 TI - [Myocardial scintigraphy. Clinical use and consequences in the county of Aarhus]. PMID- 14752994 TI - [Compliance with the treatment of osteoporosis with diphosphonates. A questionnaire among postmenopausal women]. PMID- 14752995 TI - [Fever in pregnancy and risk of fetal death. Results from the Better Health for Mother and Child-project]. PMID- 14752996 TI - [Picture of the month: synostosis between talus and calcaneus]. PMID- 14752997 TI - [Answer to a debate: on prenatal diagnosis (Ugeskr Laeger 2003;165:4981-2)]. PMID- 14752999 TI - [Comments to the brochure "Genetic tests and drug monitoring" distributed by the Laboratoriet, Epilepsihospitalet in Dianalund]. PMID- 14752998 TI - [Professionalism in psychiatry is essential--support to a contribution by professors in psychiatry]. PMID- 14753000 TI - [Reformation of the sanatorium-health resort complex and regulation of the market of sanatorium health care services--task for all participants of the market]. PMID- 14753001 TI - [Aromatherapy in the control of autonomic regulation of the heart rythm]. AB - Significant positive changes of heart rate variability, especially heart rhythm (HR) spectral structure, were observed in patients with disturbed autonomic HR control and high anxiety level given aromatherapy according to a special program. PMID- 14753002 TI - [Mechanisms of adaptation effect of normobaric hypoxic therapy]. PMID- 14753003 TI - [Rehabilitation of patients with chronic bronchitis and spine diseases]. AB - Correction of disturbed respiratory function and inhibition of progression of pulmonary inflammatory and obstructive processes is the aim of respiration rehabilitation in patients with chronic bronchitis (CB). It is known that rehabilitation in CB patients with spinal diseases meets with difficulties as dystrophic lesions of the spine entail its functional-anatomic disorders at the corresponding level. By supporting progression of dystrophic spinal and osteovertebral junction disorders, chronic bronchopulmonary inflammation participates in formation of interrelated respiratory affections. New approaches to rehabilitation of CB patients with spinal lesions lie in use of pharmacopuncture, acupuncture, deep reflex-muscular massage and therapeutic exercise in consensual sequence. PMID- 14753004 TI - [Combined use of electric sleep and hofitol in the treatment of primary chronic gastroduodenitis]. AB - Primary chronic gastroduodenitis (PCGD) accounts for 60-85% of the diseases of the gastroduodenal zone. In our study 90 patients with PCGD were divided into three groups getting one of the following therapies: electric sleep, hofitol, electric sleep plus hofitol. The effects of the treatments were assessed with updated techniques including computed pH-metry. Hofitol showed a good effect on dyspepsia, enhanced the alkalizing ability of the duodenal bulb. Electric sleep relieved pain and asthenoneurotic syndromes, decreased high acidity of the gastric juice in the body of the stomach. Electric sleep in combination with hofitol normalized macroscopic picture of the upper gastrointestinal tract and corrected imbalanced immunity. PMID- 14753005 TI - [Efficacy and optimisation of regiments of sulfate-sodium mineral water drinking in children with biliary pathology]. PMID- 14753006 TI - [Effect of radon baths of various concentrations on patients with genital endometriosis]. AB - The study of the effect of radon water in different concentrations (1.5 and 6.5 kBk/l) on a clinical status and endocrine system of patients with genital endometriosis has detected a positive action of both concentrations. High concentration radon water more effectively decreased the size and tenderness of endometriotic foci as well as concentration of sex steroids. Therefore, a differential approach is needed to prescription of radon water. Its concentration must be based on endometriosis extension and severity of the process. PMID- 14753007 TI - [Peloid therapy in the complex sanatorium treatment of children of early age with complications of perinatal brain damage]. AB - The search for novel approaches to multimodality prophylaxis and treatment of sequelae of perinatal nervous system affection as well as introduction of the early diagnostic criteria are topical problems in present-day pediatric neurology. Peloid therapy efficacy in combined sanatorium treatment was studied (Peloterm unit) in 44 infants aged 1 to 3 years. They suffered from sequelae of perinatal affections of the central nervous system including infantile cerebral paralysis. A positive effect (improvement of motor and psychic-speech development) was achieved in 98% cases. This indicates validity of this method in the treatment of CNS affections following perinatal affection of the brain in infants over 1 year of age. PMID- 14753008 TI - [Treatment of children and adolescents with attention deficit hyperactivity syndrome using transcranial micropolarization]. PMID- 14753009 TI - [Radioprotective effect of drinking sulfate mineral water on spermatogenesis in offspring of irradiated male rats]. AB - Histological and electron-microscopic studies of a radioprotective action of drinking sulphate mineral water (SMW) on spermatogenesis of irradiated male rats' progeny have found that SMW used before radiation (2 Gy) and 10 days after it is able to reduce postradiation sequelae in the progeny (2-5 month and 1.5 year old rats) testes. PMID- 14753010 TI - [Effect of boron-containing mineral water on glucose tolerance in rats with experimental prediabetes]. PMID- 14753011 TI - [Endourethral collalysine electrophoresis in the treatment of chronic prostatitis]. PMID- 14753012 TI - [Effect of iodobromine baths on some risk factors of ischemic heart disease]. PMID- 14753013 TI - [Laser infrared irradiation in the complex treatment of gastroduodenal ulcer]. PMID- 14753014 TI - [Rehabilitation of the myofascial disfunction of nodding muscles in sanatorium]. PMID- 14753015 TI - [Osteochondrosis and arthrosis in clinical practice]. PMID- 14753016 TI - [Role of air of the working areas in the development of fatigue and rehabilitation of health reserves of computer operators]. PMID- 14753017 TI - [Optimization of sanatorium-health resort care for employees of gas industry]. PMID- 14753018 TI - [Evgenii Andreevich Larin. A worthy example of serving the Caucasus Mineral Water resort]. PMID- 14753019 TI - [Problems of radon therapy in USA]. PMID- 14753020 TI - [The status of radon resort should be returned to Belokurikha]. PMID- 14753021 TI - Medical management of the healthcare worker with conjunctivitis. PMID- 14753022 TI - Effectiveness of interventions influencing activities of daily living in visually impaired patients: a systematic review. PMID- 14753023 TI - Pursuing passionate professionalism: the journey of one ophthalmic nurse. PMID- 14753024 TI - Low vision rehabilitation: a concise overview. AB - This article provides a concise overview of low vision, evaluation and rehabilitation. PMID- 14753025 TI - [Nocturnal traffic noise and stress hormone elevations in children]. PMID- 14753026 TI - [Effects of long-term exposure to street traffic exhaust on the development of skin and respiratory tract diseases in children]. AB - The pathogenesis of allergies can be stimulated by adjuvant effects--i.e. air pollutants such as NOx and particles from diesel engines as well as noise--the latter especially during night-time. During sleep, noise signals which are associated with danger (i.e. lorry noise) have the potential to trigger stress reactions even if the noise level is low. Increases of cortisol in the first half of the night seem to play an important role.--In a blind interview study, the combined effects of chronic exposure to traffic related air pollution and noise, upon the risk of skin and respiratory diseases in children were studied. All children between 5-12 years, who had consulted one of two participating pediatricians were included in the study. The pediatricians' diagnoses of 400 children were analysed together with their parents answers on the density of road traffic on their street and several confounding factors. Multiple regression analyses resulted in relative risks of asthma, chronic bronchitis and neurodermitis, which increased significantly with increasing traffic load. A comparison with the literature on such effects caused by air pollution alone, showed that traffic noise during the night might have an adjuvant effect on the pathogenesis of the mentioned diseases. PMID- 14753027 TI - [Respiratory tract diseases in children stressed by traffic noise and automobile exhaust--a field study]. PMID- 14753028 TI - [Epidemiologic reasons for screening programs in the national health service]. AB - The author describes the current health state of the Hungarian population in terms of cancer mortality and morbidity. Based on the comparative analysis of national and international, mainly European, data he describes the unfavourable Hungarian indices trying to identify their causes and the possible breaking free from them, as well. The greatest potential lies in the organised, continuous screenings within the frame of "Johan Bela National Programme of the Decade of Health". Since tumour diseases pose severe and alarming problems in national health care the reduction of extremely high mortality in three tumour localisation (cervix uteri, breast and colorectum) by regular screenings is absolutely justified. PMID- 14753030 TI - [Epilepsy surgery in childhood: theory and practice]. AB - 1-1.5% of the Hungarian child population has epilepsy, and around 20% of them produces seizures in spite of modern antiepileptic drugs. A part of the pharmaco resistant children may benefit from surgical removal of the epileptogenic focus. Presurgical evaluation has been developed a lot since the progress of neuroimaging, video-EEG monitoring, neuropsychology, and neurosurgical techniques in the 1990s. Authors summarize the important steps of presurgical evaluation in epileptic children emphasizing the role of history taking, physical examination, neuroimaging, standard EEG, long term monitoring, and neuropsychology. They describe the surgical treatment of the most important epilepsy syndromes in childhood analyzing the data of 58 epileptic children examined in the Bethesda Children's Hospital and operated in the National Institute of Neurosurgery (Budapest, Hungary). Age of children at surgery was between 16 months and 18 years, dysplasia and benign tumors covered etiology in 59% of the cases. The most frequent intervention was resection; however also some callosotomies and hemispherotomies were completed. Authors emphasize the importance of early surgical intervention of therapy resistant children in order to prevent the deteriorating effects of epilepsy on childhood psychomotor development. PMID- 14753029 TI - [Management of postoperative inflammatory response and pain with nimesulide after cardiac surgery]. AB - INTRODUCTION: Nimesulid (Mesulid) is a non-steroid anti-inflammatory drug (NSAID), acting by the selective inhibition of the Cyclooxygenase-2 (COX-2) isoenzyme. In this study the efficacy of nimesulid following cardiac surgery has been investigated in comparison with that of a COX-1 isoenzyme inhibitor drug. PATIENTS AND METHODS: 200 cardiac surgical patients operated on cardiopulmonary bypass have been involved in this prospective study. 100 patients received 100 mg. nimesulid bd. routinely in the postoperative period (group M). Another 100 patients were given 275 mg naproxen bd. (group A). In both groups the White Blood Cell (WBC), the blood sedimentation (We), the C-Reactive Protein (CRP) levels were determined from venous blood samples before the operation and on the first five postoperative days. Venous blood samples of 20 patients of each group in the same period were used to determine the Interleukin-6 (IL-6), and the Soluble Tumour Necrosis Factor Receptor-I (sTNF-RI) levels. Postoperative complications related to the use of the investigated drugs as well as the side effects have been compared in the two groups. A visual analogue pain scale was used before and after drug administration. Kruskal Wallis and student t tests were used for the statistical analysis. RESULTS: No serious complication related to the use of the investigated drugs has developed in either group. In group A gastrointestinal side effect were recorded in 7 cases (7%) whilst in group M no such complaints were found. Drug doses defined in the study protocol had to be raised or another drug had to be added in 11 and 3 cases in group A and M respectively. Neither the laboratory findings, nor the visual pain scale results have not shown any significant difference between the groups. CONCLUSION: The efficacy of nimesulid in postoperative inflammatory inhibition and pain relief has proved equal to that of the investigated COX-1 inhibitor drug, however less gastrointestinal side effects have been related to its administration. PMID- 14753031 TI - [Combined interferon-alfa-2b and ribavirin therapy in patients with recurrent chronic hepatitis c after liver transplantation]. AB - Interferon with ribavirin therapy has been proposed for the treatment of hepatitis C recurring in liver transplants. AIM OF THE STUDY: Was to assess the efficacy of standard combination therapy (interferon plus ribavirin) of chronic hepatitis C in transplanted patients with recurrent severe HCV induced chronic hepatitis. METHODS: 12 patients with HCV-PCR positive reaction (genotype 1b) were treated with the therapy of interferon-alpha-2b (3 MU three times a week) and 800 1000 mg ribavirin daily. Liver biopsy had been done in every patients before and after the treatment. Study endpoints were the end of treatment and the 6 month post-therapy sustained virologic response. RESULTS: At the end of treatment 3 patients were negative for HCV-PCR and all of them had negative reaction after 6 month follow-up period. CONCLUSION: The results are in a good accordance with treatment of patients with chronic hepatitis C without liver transplantation. PMID- 14753033 TI - [Hospital founding in the besieged capital of Hungary]. PMID- 14753032 TI - [Treatment and prevention of infectious endocarditis]. PMID- 14753034 TI - [Gyula Veszi (1888-1918)]. PMID- 14753035 TI - Twin Force Bite Corrector--hyper efficient Class II correction for a busy orthodontic practice. PMID- 14753036 TI - The Crown Bite Jumping Herbst. AB - The Crown Bite Jumping Herbst Appliance is evaluated and combined with Straight Wire Arch Fixed Orthodontics in treatment of Class II, Division I malocclusions. This article will evaluate a combined orthodontic approach of "straightening teeth" and an orthognathic approach of "moving jaws or making skeletal changes." Orthodontic treatment cannot be accomplished well without establishing a healthy temporomandibular joint. This is defined by Keller as a joint that is "noiseless, painless and has a normal range of motion without deviation and deflection." It is not prudent to separate orthodontic treatment as its own entity without being aware of the changes in the temporomandibular joint before, during and after treatment. In other words, "If you're doing orthodontics you're doing TMJ treatment." One should treat toward a healthy, beautiful face asking, "Will proposed treatment achieve this goal?" Treatment should be able to be carried out in an efficient manner, minimizing treatment time, be comfortable and affordable for the patient, and profitable for the dentist. The finished treatment should meet Andrews' Six Keys of Occlusion, or Loudon's Twelve Commandments. Above all, do no harm to the patient. We think that a specific treatment plan can embrace these tenets. The focus will be to show Class II treatment using a modified Herbst Appliance and fixed straight wire orthodontics. PMID- 14753037 TI - Ortho hints. Initial wire selection.. PMID- 14753038 TI - Ortho hints. What is your starting wire? PMID- 14753039 TI - [Appeal of 2d Russian "Occupation and Health" Congress participants to Russian Federation's President V V. Putin]. PMID- 14753040 TI - [Health of worker is a topic of national security]. PMID- 14753041 TI - [Social responsibility of the employer for the employees' health and contemporary role of occupational medicine]. AB - Changes taken place over past 15 years in economic and social life of Russia have significant negative influence on workers' health, recovery of labour resources. Sharing responsibility for those problems solution between state and society, employers and employees, medical science and practice is one of the most topical problems. PMID- 14753042 TI - [Problems and prospects of physicians' training in "medical prophylaxis" specialty]. AB - According to a Concept of continuous special education, postgraduate special training remains an essential link in creating a widely educated doctor with humanistic and natural-science ideology, high culture and knowledge of contemporary circumstances. PMID- 14753043 TI - [Social and hygienic monitoring of mining enterprises]. AB - The authors justified parameters to provide social and hygienic monitoring of work conditions at mining enterprises. PMID- 14753044 TI - [Effects of work at video displays on the state of the nervous system]. AB - Videodisplay users were compared with reference group in nervous system state, length of service (under 5 years, 5-10 years, 10 years and more) during prospective observation. The comparison demonstrated reliably high and occupationally induced risk of vegetative dysfunctions. The authors defined correlation of the risk with exposure to 2-400 kHz magnetic fields. PMID- 14753045 TI - [Genotoxic effects in workers engaged in ore mining and processing]. AB - The authors studied frequencies of chromosomal aberrations (CA) in 48 ore mining and processing workers exposed to occupational hazards complex. Finding is significantly increased frequency of CA, when compared to that in reference groups. Level of structural CA in the main group appeared not to depend on age, sex and bad habits (smoking). The study revealed indirect dependence between cytogenetic disorders frequency and length of service. PMID- 14753046 TI - [Occupational stress (literature review)]. PMID- 14753047 TI - [Forecasting harmless service and work safety for able-bodied population]. AB - The authors suggest medical and mathematical simulator for forecasting hygienically harmless length of service with consideration of occupational load and other circumstances causing human wear and ageing. Forecasting length of individual occupational longevity is a way to novel system of work safety and health preservation for able-bodied population. PMID- 14753049 TI - [Atomic absorption for determining selenium in biological material]. AB - To determine selenium with high sensitivity, the authors suggest atomic absorption based on selenium hydride formation and thermal decomposition with further selenium line absorption measurement. Low limit of selenium determination in an analysed sample is 5 ng, with standard error no more than 10%. PMID- 14753048 TI - [Hygienic evaluation of work conditions, health status and determination of occupational risk for engineers and technicians in civil aviation]. AB - The article deals with hygienic evaluation of work conditions, health state and determining occupational risk for engineers and technicians servicing aircrafts. The main hazards, according to the authors, are work intensity, high noise levels, air pollution with chemicals and unfavorable microclimate. A complex of prophylactic measures was justified. PMID- 14753050 TI - [Comparison of maximal physical load tolerance in rescuers working with insulation respiratory devices using air and oxygen-helium mixture]. PMID- 14753051 TI - [Changes in dust content of workplace air during ore extraction at the Bazhenovsk chrysotile asbestos field]. PMID- 14753052 TI - Reshaping Medicare--prescription drug coverage for elderly adults. PMID- 14753053 TI - What's new about old drugs. AB - Every month new clinical trials are published that provide relevant insight into medical care. Health care professionals are expected to review the results of these trials to update their knowledge and clinical practice. Although it is impossible to review every clinical trial, it is important to evaluate study findings in one's area of interest or practice. For nurses and other practitioners in the field of geriatrics, clinical trials involving drug therapy can be particularly valuable. This article is a review of clinical trials published in the past year that provide new information about drug therapy used by elderly patients. It reviews recent clinical trials in the areas of cardiology (hypertension, dyslipidemia, antioxidants for cardiovascular disease, hormone replacement therapy, atrial fibrillation, systolic heart failure), hematology (venous thromboembolic disease), neurology (Parkinson's disease, post-herpetic neuralgia), and rheumatology (osteoarthritis). Major findings and implications for geriatric clinical practice are included. PMID- 14753054 TI - Anxiety and nighttime behavioral disturbances. Awakenings in patients with Alzheimer's disease. AB - This study was conducted to describe the relationship between anxiety and nighttime behavioral disturbance in a community-dwelling sample of patients with Alzheimer's disease (AD). Data from 153 patients with probable or possible AD and their family caregivers were analyzed using logistic regression modeling. Ratings of nighttime behavioral disturbance were based on caregiver reports of how often patients had awakened them at night during the past week. Standardized ratings for patient cognitive, functional, and behavioral status, and for caregiver sleep, depression, and burden were collected. Fifty-six percent of the patients with AD showed symptoms of anxiety, and 29% had awakened their caregiver at least once at night during the past week. Patient awakening was associated with higher levels of patient anxiety (odds ratio [OR] = 2.1; confidence Interval [CI] = 1.4, 2.9) and patient impairments in activities of daily living (OR = 1.6, CI = 1.2, 2.3). No other demographic, cognitive, functional, or behavioral variables were significant, including depression. In univariate analyses, individual patient anxiety symptoms (e.g., feeling anxious; showing physical signs of anxiety, agitation, and irritability) were significant risk factors for patient awakenings. Of these, showing physical signs of anxiety remained a significant risk factor in multivariate analyses. Results suggest that anxiety and nighttime awakening are highly interrelated in patients with moderate dementia due to AD, and treatments targeting both may be more efficacious than those focusing on anxiety or sleep alone. They also reveal the importance of assessing anxiety as well as depression in the research and clinical care of patients with AD. PMID- 14753055 TI - Shifting paradigms. Teaching gerontological nursing from a new perspective. AB - Gerontological health care is undergoing a revolution, much like that of the feminist movement of the 1960s. Fundamental changes in health care require revisions in nursing education to ensure appropriate care of older adults in the least restrictive environment. The purpose of this study is to promote the preparation of future nurses who have the knowledge and the skills necessary to provide nursing care for the growing cohorts of older adults. A theoretical rationale for a new perspective in nursing education is discussed. An experiential clinical learning activity based on the functional model of gerontological health care is examined. This home visit clinical learning activity provides nursing students with the opportunity to practice nursing reflective of the health care needs of older adults. Strategies for replication of this clinical learning activity are provided. PMID- 14753056 TI - Maintaining the square. How older adults with Parkinson's disease sustain quality in their lives. AB - In this article, the author's objective is to uncover the common practices that help sustain quality in the lives of older adults with Parkinson's disease. Interpretive phenomenology was the method used to gather and analyze the stories of six men and six women with idiopathic Parkinson's disease. The results show a constitutive pattern of Maintaining the Square. Five relational themes, including Learning How, Accepting Limitations, Seeking Knowledge, Engaging in Meaningful Experiences, and Living for Today, emerged from the participants' stories. The participants' ability to sustain continuity in their lives indicates their successful maintenance of the square. By identifying markers of continuity, they are able to maintain quality in their lives. As nurses, we can promote continuity in the lives of our patients if we take the time to learn what gives them meaning and purpose in life and then facilitate ways to help them maintain this connection. PMID- 14753057 TI - Understanding nutritional health in older adults. A pilot study. AB - A pilot study of adults ages 65 and older admitted to an acute care setting was conducted to compare nutritional risk as measured by hospital dieticians with two Nutrition Screening Initiative tools, the DETERMINE Your Nutritional Health Checklist and the Level I Screen, and to elicit from patients their own perceptions of nutritional health. Ten community-living older adults were interviewed. Although all 10 were at nutritional risk as measured by both hospital assessment and nutritional risk screening tools, none of these patients believed themselves to be at risk. One conclusion of this pilot is that interventions and education need to be tailored to the perceptions of targeted individuals. PMID- 14753058 TI - Blood pressure monitoring. Older adults' perceptions. AB - The purpose of this article is to report the qualitative findings on older adults' perceptions of blood pressure measurement and its meaning to their health. The convenience sample consisted of 51 community-dwelling older adults who were recruited from either wellness clinics or a senior citizens' club. Data were collected through in-depth audiotaped interviews. Participants' responses were analyzed by identifying topics, patterns, and themes. The following three themes emerged: provision of reassurance, decision-making leading to self-care activities, and reliance on health care professionals. These findings can be used by community health nurses, especially those in gerontology to gain insight into how a sample of older adults perceived blood pressure monitoring. In addition, the findings can serve as guidelines for nurses in the provision of effective hypertension management with older adults. PMID- 14753059 TI - Smoothing the pathways. PMID- 14753060 TI - Making the right connections. PMID- 14753061 TI - Positively young at heart. PMID- 14753062 TI - Be aware of the benefits and risks of bed-sharing. PMID- 14753063 TI - Be aware of the benefits and risks of bed-sharing. PMID- 14753065 TI - Stress, burnout and trauma in health care. When working hurts. PMID- 14753066 TI - Pharmaceutical waste and nursing practice. Interview by Helen Griffiths. PMID- 14753067 TI - Joy Millis' new vision for dental implant practice. PMID- 14753068 TI - Models and evidence-based practice: you can't have one without the other. PMID- 14753069 TI - Theory of research utilization enhancement: a model for occupational therapy. AB - BACKGROUND: There is a pressing need for occupational therapists to provide research-based practice, yet there is little understanding of the specific strategies and processes individual practitioners use to integrate research evidence into their clinical practice. METHOD: Using grounded theory method, the self-reported research utilization strategies of a sample of 11 elite occupational therapists practicing in adult stroke rehabilitation were examined. The triangulation of the interview data, the organizational policies of their workplaces, and existing theoretical concepts and processes of research utilization resulted in the development of a theory and a practice model to guide research utilization in occupational therapy. RESULTS: The Theory of Research Utilization Enhancement for Occupational Therapists, and the Model of Research Utilization in Occupational Therapy are presented, and their implications for practice, policy, education and future research are discussed. PRACTICE IMPLICATIONS: Built upon the Occupational Performance Process Model, the theory and model are proposed as guides to enhance therapists' ability to maintain a client-centred approach while informing clinical practices with research evidence. The application of structured reflection, case application and peer consultation facilitate the integration of research evidence into clinical practices. PMID- 14753070 TI - Occupational value among individuals with long-term mental illness. AB - PURPOSE: The study compared the perceived value of occupation among a sample of individuals with long-term mental illness to a sample of people not diagnosed with mental illness. As well, it investigated whether diagnostic and demographic factors were related to perceived occupational value among the individuals with mental illness. Finally, the study examined the relationship between occupational value and ratings of health and well-being. METHOD: One hundred and three individuals with mental illness and 28 healthy individuals were recruited for the study. RESULTS: Overall occupational value among the individuals diagnosed with mental illness differed only marginally from the healthy group, indicating that perceived occupational value was by and large not related to mental illness. Among the individuals with mental illness, having children living at home was related to occupational value. There were moderate to strong associations between occupational value and measures of health and well-being. PRACTICE IMPLICATIONS: This study provides important insights into occupational value among individuals with persistent mental health problems and provides some preliminary evidence in support of the Value, Meaning and Occupation Model. PMID- 14753071 TI - The effects of family-centred functional therapy on the occupational performance of children with cerebral palsy. AB - BACKGROUND: Family-Centred Functional Therapy (FCFT) is a comprehensive approach to paediatric occupational therapy that considers and addresses the skills of the child, the requirements of functional tasks and the elements within the tasks environment, including family goals and preferences. PURPOSE: The purpose of the present study was to determine if interventions based on FCFT, but limited to addressing elements of the task and environment, would improve task performance. METHOD: A single subject, multiple baseline design was applied to three children, for three tasks each. Tasks were selected by parents and measured during baseline and intervention phases. RESULTS: Results indicate that for each child at least one of the two tasks for which intervention was provided demonstrated improved performance. Parents reported a clinically significant improvement in satisfaction with their children's performance in the tasks that were addressed. PRACTICE IMPLICATIONS: This study provides preliminary evidence of the effectiveness of the FCFT approach. PMID- 14753072 TI - Supported employment: evidence for a best practice model in psychosocial rehabilitation. AB - BACKGROUND: Traditional approaches to vocational rehabilitation in mental health settings have had only limited effectiveness in enabling clients to meet their employment goals. Within the last decade the individual Placement and Support Model (IPS) has emerged as an alternate, evidence-based approach to providing vocational services with individuals who have severe and persistent mental illness. METHOD AND SCOPE: This review of the literature critically examines research regarding the IPS model of supported employment then discusses implications of this research for occupational therapists. PRACTICE IMPLICATIONS: In order to enable clients to achieve their competitive employment goals, it is imperative that occupational therapists incorporate best practice models of supported employment. IPS is one model that appears to hold significant promise for occupational therapists and their clients. PMID- 14753074 TI - Networking and communication for nurses. PMID- 14753080 TI - Obesity: a word that needs further discussion. PMID- 14753081 TI - Telemedicine at Huntertown Elementary, Versailles, Kentucky. PMID- 14753082 TI - Body piercing. AB - A student comes to your office excited about the prospect of getting her tongue pierced. "Isn't it cool?" The popular trend of body piercing is on the rise. It is our job as nurses and health promoters to alert students to the facts. Students must be informed about the risks involved so they can make suitable decisions about their bodies. Piercing may be more than an assertion of individuality. This article describes body piercing health risks and treatment methods, and how school nurses can advise students on the risks and care of body piercing. PMID- 14753083 TI - Sugar shock: limiting sugar intake for the sake of oral health is tougher than it sounds. PMID- 14753084 TI - Update on type 2 diabetes in children and adolescents. PMID- 14753085 TI - Resiliency-based research and adolescent health behaviors. PMID- 14753086 TI - Making chicken pox prevention a priority. AB - School nurses are the IDEAL professionals to catch those kids without a history of chicken pox (varicella illness) who have not been immunized, thereby falling into the susceptible range. We handle charts daily. We have both illness history (if it was provided) and immunization dates. We can call parents. We can talk to kids. Wouldn't it be easy if we could do that and then call (or email) the primary care provider? But there is no longer an "easy" way to communicate health information. Doing things the easy way could be perceived by some as "tattling" and by others as "reporting without permission." Instead, the school nurse must take the time-consuming (and better long range) path of education ... of the student, and the parent. And the priority can slip down below other school nurse requirements such as getting kids their required immunizations, first aid and illness care, individualized health care plans for students with medical problems, parent and teacher phone calls, and endless conversations. It takes a village to raise a child though, and the school nurse is a village resident. The following information is vital to the knowledge base of today's school nurse. Please read it, store it and consider it when planning and intervening with your students in the day-to-day interaction that promotes optimal health and wellness. PMID- 14753087 TI - Anaphylaxis/acute allergic emergencies. PMID- 14753088 TI - Internet information that's free, accurate and helpful. PMID- 14753089 TI - Do you know what your patients are reading? PMID- 14753090 TI - Screening for violence in pregnancy. PMID- 14753093 TI - Is emergency contraception going over-the-counter? PMID- 14753094 TI - Evaluating the female pelvic floor. PMID- 14753095 TI - Mifepristone for medical abortion. Exploring a new option for nurse practitioners. PMID- 14753096 TI - Newborn stimulation preventing over-stimulation is key for optimal growth & well being. PMID- 14753097 TI - Facing stillbirth or neonatal death. Providing culturally appropriate care for Jewish families. PMID- 14753098 TI - Circle of life. PMID- 14753099 TI - Hemodialysis adequacy and quality of life: how do they relate? AB - There is clear evidence of a link between dialysis adequacy (as measured by urea kinetic modelling or urea reduction ratio) and such important clinical outcomes as morbidity and mortality. Evidence regarding the relationship between dialysis adequacy and quality of life outcomes is less clear. This paper reports the results of a study designed to answer the following research question: What is the impact of changing dialysis adequacy on functional status and well-being in a sample of new hemodialysis patients? The study employed a convenience sampling technique enrolling all willing new patients who started hemodialysis for a four year period. The final sample size was 69. The adequacy of each new patient's dialysis prescription was determined by calculating Kt/V at one and three months following the start of dialysis. Changes to dialysis prescriptions were made, as necessary, to achieve a minimum Kt/V of 1.2. Each subject's quality of life was measured at the same time intervals, using two instruments. The first instrument, the SF-36, is a generic 36-item instrument designed to measure a range of functioning and well-being. The second instrument, the Kidney Disease Questionnaire (KDQ), is a disease-specific quality of life instrument designed for use with chronic hemodialysis patients. The results of this study demonstrated a significant positive relationship (p < .05) between Kt/V and four of five variables in the KDQ, and two of eight variables in the SF-36. These variables were predominantly in the social/emotional well-being domain. PMID- 14753100 TI - Dilemmas in providing patient-focused care. AB - The provision of patient-focused care to the elderly is often fraught with many ethical and challenging dilemmas. However, for health care professionals, these dilemmas can be magnified when the chronic disease is accompanied by dementia. Such was the case of Mrs. A. The multidisciplinary team of a 36-bed acute care unit identified the challenges they experienced while caring for a 65-year-old female with multiple medical problems: diabetes; hypertension; cardiovascular disease; end stage renal disease and dementia probably due to cardiovascular and metabolic causes. Mrs. A. was admitted to start hemodialysis treatments. Her care was complicated by frequent outbursts of verbal and physical aggression towards staff. Mrs. A. was deemed incapable with regard to personal care and property. Her son believed she required long-term placement, and control of her property was assumed by the Public Guardian and Trustee. Mrs. A. vehemently objected to this total loss of control. This paper will describe how the multidisciplinary team implemented a variety of strategies to help staff intervene more effectively in meeting the care needs of Mrs. A. PMID- 14753101 TI - Proteinuria, a modifiable risk factor: angiotensin converting enzyme inhibitors (ACEIs) and angiotensin II receptor blockers (ARBs). AB - Microalbuminuria and proteinuria have been determined to be modifiable risk factors for the progression of chronic kidney disease as well as risk factors for cardiovascular events. Angiotensin converting enzyme inhibitors and angiotensin II receptor blockers have been demonstrated to decrease proteinuria at all stages and slow the progression of renal disease. Proteinuria can be used as a marker of successful treatment in patients with chronic kidney disease in combination with other established targets. This article discusses the various diagnostic tests used for the detection of microalbuminuria and proteinuria and appropriate pharmaceutical treatment. PMID- 14753102 TI - Timing and effect of fissure sealants on permanent molars: a study in Finland and Sweden. AB - Fissure sealants are frequently used in the Nordic countries for the prevention of caries in children. The aim here was to analyse their use and effect on dental health in children and adolescents in Finland and Sweden. A retrospective longitudinal population-based study was performed in northern Finland and Sweden, where different strategies have been adopted for sealing teeth. The timing of sealant and restoration placements up to 18 years was studied in representative samples of subjects born in the early 1980s in Kemi, Oulu, Raahe and Tornio in Finland (n = 908) and compared with a random sample of subjects from Norrbotten, Sweden (n = 634). A sensitive survival analysis method was applied to all the permanent molars separately (Virtanen et al. 1996). About 80-90% of molars had been sealed in the Finnish subjects, and approximately 30% in the Swedish cases, employing selective criteria. The filling increments were statistically significantly higher in Sweden than in Finland, although the differences were not of the same magnitude as the reduction in sealants. More than 30% of first molars and 50% of second molars had not been treated, i.e. were without sealants or fillings, by the end of the follow-up in Sweden, compared with 10% and 20%, respectively, in Finland. Fissure sealants were effective in preventing caries in molars in children and adolescents, and the use of selective criteria for targeting this treatment can yield savings in effort and costs in a low caries situation, although more research is needed to find the optimum cost-effective strategy. PMID- 14753103 TI - Bonded resin sealant on smooth surface dental enamel--an in vitro study. AB - The aim of this study was to in vitro evaluate the micro leakage of a fissure sealant on sound and demineralized enamel using thermo cycling. The effect of the sealing procedure on sound and demineralized enamel were assessed on human premolars that were divided into one Control group and two Test groups. The Control group served as a baseline reference for the subsequent test procedures. The crowns of five teeth were covered with wax leaving three separate windows on the buccal surfaces. The enamel in one window was acid-etched, in the second window demineralized, and in the third window demineralized and subsequently acid etched. In Test group 1, containing ten teeth, a defined enamel area on the buccal surfaces was acid-etched and sealed. The sealed area and a border of sound enamel around it were subjected to the demineralizing procedure. In Test group 2, a defined enamel window of 13 teeth was demineralized and subsequently etched and sealed. The sealed area and a border of surrounding enamel were subjected to a new cycle of demineralization. The longitudinal, bucco-lingual sections were examined in polarized light. When placed on acid-etched surfaces, the resin adhered firmly to the enamel with no evidence of demineralization or enlargement of previously demineralized areas underneath the sealants. PMID- 14753104 TI - Changes in tobacco habits. A prospective longitudinal study of tobacco habits among boys who play ice hockey. AB - The aim of the investigation was to follow up tobacco habits and actual sporting activities among the boys who participated in an earlier study by the authors, and to examine whether knowledge of the harmful effects of tobacco and tobacco behaviour patterns and/or the choice of tobacco had changed amongst the participants. The study was conducted 3 years after the original study. Data were collected using a questionnaire. Of the 183 boys in the age group 15-22 years who participated in the study, 26.8% were tobacco users: 19.7% only snuff users, 6.0% both snuff users and smokers and 1.1% smokers. Compared with the original study, a further 16.4% of the boys had started to use tobacco. The use of tobacco increased in all age groups except amongst 19-year-olds where tobacco use was unchanged. The age group 17-22 years included boys who were both snuff users and smokers. An increase in tobacco use between the ages of 17 and 19 years could be shown compared with the original study, for equivalent age groups. Among the 132 boys who still played ice hockey, 25.8% were tobacco users: 18.2% only snuff users, 6.1% both snuff users and smokers and 1.5% smokers. A statistically significant difference (p < 0.05) could be shown between boys who used snuff and non-users depending on whether the boys participated in another sport besides ice hockey. Snuff usage was thus not as widespread among boys who participated in other sporting activities. The increase in tobacco use amongst the boys in the study showed that the various preventive initiatives which society so far has offered have been inadequate. Hence, the preventive measures need to be evaluated and developed to prevent tobacco habits from becoming established. The results of this and previous studies show that the environment in which ice hockey is practiced can, in itself, constitute a risk for tobacco usage becoming established among ice-hockey-playing adolescents. PMID- 14753105 TI - Oral disease and psychosocial risk determinants in relation to self-assessments of general health in persons with chronic whiplash-related disorders. AB - The aim of this study was to analyse how self-assessed general health was related to oral health among persons afflicted with whiplash-associated disorders (WAD), controlling for relevant background factors, confounders and other risk factors. Questionnaires included the SF-36 Health Survey, self-assessed oral health and relevant risk factors, in total 49 questions. Multivariable regression modelling was performed. Members of a nationwide Swedish association enlisting persons who have problems concerning a whiplash injury (n = 1,928) were included. A total of 979 persons participated in the study, a response rate of 50.8%. A multivariable regression model is presented, with general health as the dependent variable, and the independent variables inserted en-bloc. The model was highly significant with an explained variance of 28%. Among background factors, only older age appeared as significantly and strongly related to poorer general health. The strongest explanatory contributions came from the health related variables. Oral disease and extra-oral body pain were both strongly related to poorer general health, most obviously for the oral disease variable. Oral disease was significantly and to a clinically relevant degree associated with self-assessed general health. Several other psychosocial indicators of stress were also significantly related to the general health. These findings are consistent with the stress-behaviour immune model for development of disease. PMID- 14753106 TI - [A new orientation to the pharmacy compounding in Belgium]. PMID- 14753107 TI - [Considerations of Erasmus on medicine, medicines, disease and pharmacopoeias of the time]. PMID- 14753108 TI - [The nurse and the disabled person... a community]. PMID- 14753110 TI - [Lumbar disk hernia -- nursing approach]. PMID- 14753109 TI - [Training... qualifying... nurses and organizations]. PMID- 14753111 TI - [The nurse: delivery of holistic care]. PMID- 14753112 TI - [What is the future of stem cells?]. PMID- 14753113 TI - [Computers: do they help or harm the care for the patient?]. PMID- 14753114 TI - [Physical activity and type II diabetes]. PMID- 14753115 TI - Local fluoridation moves a step nearer. PMID- 14753116 TI - Postnatal depression: an overview. PMID- 14753117 TI - Health surveillance in childhood: a debate. PMID- 14753118 TI - Screening: we should welcome a more rigorous approach. PMID- 14753120 TI - Health and fitness series--6. Child health: how nutrition affects growth. AB - Dr Margaret Lawson summarises the relationship between childhood growth and nutrition, and the effect of early nutrition on health in later life. She explains why children should be encouraged to eat a varied diet that includes foods from each of the four main food groups. The need to monitor children's growth is discussed, as are some of the nutritional pitfalls that may prevent children achieving their optimum growth and health. PMID- 14753121 TI - On target for health. PMID- 14753122 TI - Want to give up smoking? Self-help tips for professionals. AB - Health professionals are well aware of the health risks of nicotine but find it as hard as lay people to give up smoking. How people prepare themselves for quitting is important in determining success. Jennifer Percival, Tobacco Education Project Manager at the Royal College of Nursing, explains how to assess motivation and measure one's level of addiction. She also gives practical steps for getting off to a successful start. PMID- 14753123 TI - ['Palm reading' as a diagnostic aid]. AB - In four patients, a man aged 53, a woman aged 63 and two men aged 67 and 53 years, respectively, inspection of the hand led to the final diagnosis. In the first patient who presented with clubbing of the fingers and pulmonary hypertension, a small atrial septal defect was detected. The second patient had thick curved yellow nails and recurrent pleural effusions as part of this yellow nail syndrome. The third patient presented with clubbing and hypertrophic osteoarthropathy, a secondary complaint to adenocarcinoma of the lung. The fourth patient had increasing pulmonary hypertension as a secondary complaint to recurrent multiple pulmonary embolism from deep venous malformations of his right hand, arm and shoulder as a part of the Klippel-Trenaunay syndrome. These cases show that inspection of the hand may give a clue to the diagnosis. PMID- 14753124 TI - [The practice guideline 'Pregnancy and puerperium' (first revision) from the Dutch College of General Practitioners; a response from the perspective of general practice medicine]. AB - The first revision of the Dutch College of General Practitioners' practice guideline about pregnancy and puerperium does not significantly differ from the first edition. The guideline is extensive, is well-worth reading and supports daily practice. There is a greater emphasis on the importance of cooperation and differentiation in primary care (midwifes and general practitioners). During the last decade many general practitioners stopped doing home deliveries and have therefore lost their experience in obstetric care and pathology. The guideline describes the general practitioner's tasks as a preconception counsellor, a professional expert on illnesses during pregnancy and after the delivery, and as the doctor of the newborn baby. It will hopefully stimulate a revived interest of and involvement in pregnancy and post-partum care among general practitioners. PMID- 14753125 TI - [The practice guideline 'Pregnancy and puerperium' (first revision) from the Dutch College of General Practitioners; a response from the perspective of gynaecology]. AB - The major benefit of the revised version of the Dutch College of General Practitioners' practice guideline about pregnancy and puerperium is the formal starting point of professional equality of midwives and general practitioners as care providers in antenatal and postpartum care. This thorough evidence-based practice guideline is behind the most recent developments in this field. Scientific evidence on vertical HIV transmission favours screening all pregnant women and not just the selective procedure proposed in this guideline. The restrictive attitude towards routine ultrasound screening during pregnancy hardly seems in line with scientific evidence and the public demand. On the other hand the guideline is very progressive as far as the care of women with thyroid disorders is concerned even though evidence on this subject is meagre. Most importantly the transition of antenatal care from the current almost conveyor belt-like procedure to an efficient but individually-adapted care process was not addressed. PMID- 14753126 TI - [Treatment of optic neuritis]. AB - Optic neuritis manifests itself as a reversible loss of vision. It can be a clinically isolated incident or one of the (first) manifestations of multiple sclerosis (MS). Its differential diagnosis is extensive, and management of other disorders can differ radically. In a typical case, treatment with corticosteroids hastens recovery of vision but does not affect the eventual degree of recovery. There is a substantial risk of developing MS after isolated optic neuritis (approximately 50% within 20 years), especially if asymptomatic white matter lesions are found on MRI scanning of the brain. Intravenous treatment with methylprednisolone may delay the onset of MS somewhat, but after three years the benefit of this treatment is lost. Treatment with interferon beta-Ia in high-risk patients also slows down the progression to clinically definite MS, but the long term benefits are uncertain. MR imaging of the brain has implications for prognosis but not for treatment. PMID- 14753127 TI - [Summary of the practice guideline 'Pregnancy and puerperium' from the Dutch College of General Practitioners]. AB - During the first trimester of pregnancy, obstetric care concentrates particularly on assessment of gestational age (by history taking or if menstrual history is unclear, ultrasonography) and on the assessment of obstetric or medical risk factors necessitating a referral for specialist care. Tracing hereditary conditions in relatives is important as this is a reason for antenatal screening for congenital abnormalities. In comparison to the previous guideline, blood testing in the first trimester has been extended to include screening for irregular erythrocyte antibodies and the possibility of screening for HIV. Pregnant women with a history of thyroid problems may also be tested for serum concentrations of thyroid stimulating hormone (TSH), free T4 and, on indication, the TSH-receptor antibody levels. Since haemodilution is physiologically normal during the second half of pregnancy, at a gestational age of 18 weeks and above, only a haemoglobin-level of 6.5 mmol/l or less justifies the diagnosis of anaemia, and should treatment be commenced. If a newborn does not show any abnormalities immediately after delivery, the physical examination now routinely carried out at a few days post-partum rarely produces any additional findings and can safely be omitted. PMID- 14753128 TI - [Neuraminidase inhibitors oseltamivir and zanamivir: new means of defence against influenza]. AB - Recently two new neuraminidase inhibitors zanamivir and oseltamivir have been marketed. They appear to considerably reduce morbidity and mortality from influenza. Their adverse effects are infrequent and mild and the chance of development of pathogenically significant resistant mutants appears to be small. During the first six months of a pandemic, neuraminidase inhibitors are the only defence against the virus. It is therefore important to stockpile in each country sufficient quantities of these drugs. During the usual influenza epidemics the main value of neuraminidase inhibitors lies in their use for therapy, prophylaxis and post-exposure prophylaxis in long-term care institutions for the elderly (for prophylaxis only oseltamivir is licensed). Although data on the effectiveness against complications of influenza and on the effect on people with an increased risk of (fatal) complications as a result of an influenza virus infection are limited, the available information on the effects of the neuraminidinase inhibitors indicates that these drugs will also protect against complications and that high-risk groups will benefit from the rapid deployment of these products. The cost-effectiveness of treatment and post-exposure prophylaxis with neuraminidinase inhibitors is probably not favourable for healthy children and adults but seems to be favourable for the high-risk groups (vaccinated or not) in winter. PMID- 14753129 TI - [From gene to disease; galactosemia and galactose-1-phosphate uridyltransferase deficiency]. AB - Classical galactosaemia (Mendelian Inheritance in Man, no 230400) is an autosomal recessive disorder of galactose metabolism caused by a deficiency of the enzyme galactose-1-phosphate uridyltransferase (GALT). The GALT enzyme is responsible for the conversion of galactose-1-phosphate with UDP glucose to glucose-1 phosphate and UDP galactose. The gene encoding for GALT is located on chromosome 9p13. Patients present with hepatomegaly, liver failure, food intolerance, hypoglycaemia, muscle hypotonia, sepsis and cataract. Treatment involving the total restriction of lactose-containing foods is life-saving but many patients develop late complications such as problems of mental development, disorders of motor function, disorders of speech and hypergonadotrophic hypogonadism. PMID- 14753130 TI - [Diagnosis of pulmonary hypertension: experiences with 187 patients referred to the VU Medical Center]. AB - OBJECTIVE: To describe the results of a multidisciplinary diagnostic protocol in patients referred due to suspected pulmonary hypertension. DESIGN: Descriptive. METHOD: In 187 patients who were referred to the VU Medical Centre, Amsterdam, the Netherlands, between May 1998 and February 2003, due to suspected pulmonary hypertension, the diagnosis was established by means of a multidisciplinary diagnostic protocol. The referral and final diagnoses were analysed, as well as the treatment the patients had received. RESULTS: In 56 patients (30%), the application of the protocol lead to a modification of the diagnosis; 16 patients (9%) were found not to have pulmonary hypertension; in 20 of the 89 patients referred with 'primary pulmonary hypertension' (48% of all referrals), an underlying disease was still identified: 6 cases of a connective tissue disease, 5 cases of a chronic thromboembolic process, 5 cases of a condition of the left heart, and 4 cases of an atrial septal defect. CONCLUSION: The diagnosis 'primary pulmonary hypertension' can only be made once all secondary types of the disease have been ruled out. The application of a multidisciplinary diagnostic protocol together with a contribution of specialists with specific expertise can help to adjust the initial diagnosis and sometimes even reject it. PMID- 14753132 TI - [Progressive lymphoedema in the legs of children and adolescents]. AB - In three children, two girls aged 18 months and 15 years and a boy aged 16 years, lymphoedema of the legs was diagnosed. Investigation by dynamic lymphoscintigraphy showed no or diminished activity of lymph vessels in the lower extremities. The patients were treated by manual compression therapy and compression stockings, which controlled the oedema. Early recognition and diagnosis are important, as otherwise irreversible complications and reduced mobility may occur. PMID- 14753131 TI - [Survival of patients with aggressive non-hodgkin's lymphoma: no difference between first line treatment in a prospective randomised phase III clinical trial and first line treatment according to routine clinical practice]. AB - OBJECTIVE: To compare survival of patients with disseminated aggressive non Hodgkin's lymphoma (NHL) who were treated either as part of a clinical trial or in routine clinical practice. DESIGN: Retrospective. METHOD: The survival was studied of patients with disseminated NHL of an intermediate or high degree of malignancy who were treated in the Meander Medical Centre, Amersfoort, the Netherlands, in the years 1994-2001 with chemotherapy consisting of cyclophosphamide, doxorubicin, vincristine and prednisone (CHOP). This took place either in routine clinical practice (RCP) or as part of a clinical trial where patients < or = 60 years of age received intensified CHOP and patients > 60 years received CHOP with growth factors. Treatment data, the response to therapy, survival and prognostic factors according to the International Prognostic Index for aggressive NHL were collected by a review of the patient records. RESULTS: Fifty-nine patients were eligible for this analysis: 32 men and 27 women with a median age of 63 years (range 30-83). Of these, 35 were treated within a clinical trial and 24 were treated in RCP. The patient characteristics in the two groups were comparable. There was no difference in median survival between the trial and RCP groups, this being 27 months for all patients, 34 months for the younger patients, 20 months for the elderly patients, and 42 months for patients who achieved complete remission following chemotherapy. CONCLUSION: No difference in overall survival was found between patients with disseminated aggressive NHL who underwent treatment according to either RCP or as part of a clinical trial. It demonstrates that both patients in clinical trials and patients treated according to RCP received equally effective therapy. Recent developments in NHL treatments are promising, and therefore participation in clinical trials should be encouraged. PMID- 14753133 TI - [Health Council recommendation on the 'Contours of the basic package']. AB - As part of the planned reform of the health insurance system, the Minister of Health, Welfare and Sport has requested the Health Council to 'formulate an opinion with regard to the workable, scientifically-based criteria for identifying which health-care services should be included in a basic package'. The Committee on Contours of the Basic Health Benefit Package has now designated two sets of criteria distinguishing a 'solidarity package' from compulsory insurance. The individual 'burden of disease' combined with 'cost-effectiveness' forms a good basis upon which to define a basic package that, in accordance with the principle of solidarity (rich with poor, young with old, and healthy with sick), will be accessible to all. These criteria have, in the meantime, been applied in a number of situations. For a compulsory package, additional criteria are required: the costs of treatment, nursing or care; the extent to which the disorder that is to be prevented or treated may afflict other people; the preventive nature of services; and the impact that the use of services has on the efficiency of health care as a whole. The two sets of criteria may result in a single basic package, but a 'solidarity package' and a 'compulsory package' need not necessarily coincide. Considerations such as actuarial feasibility may have a bearing on the governmental decisions in this matter. In order to apply the criteria to different services, it is necessary to have a 'national assessment framework' that supports rational decisions. This framework will need to accommodate procedures for defining the package, since application of the criteria will always require a qualified approach. PMID- 14753134 TI - [Fitness-to-drive in neurological disorders]. PMID- 14753135 TI - [Breast cancer and the use of the hormones around the menopause]. PMID- 14753136 TI - [The road to the standardization of ambulatory anesthesia]. PMID- 14753137 TI - [What do patients think about anesthesia? Survey on postoperative satisfaction in ambulatory surgery comparing general and subarachnoid anesthesia]. AB - OBJECTIVES: To conduct a telephone survey to determine the degree of patient satisfaction with the anesthetic technique applied during outpatient arthroscopic surgery on the knee. MATERIAL AND METHODS: We recorded refusals to respond to the survey and the reasons. The patients were distributed randomly in 3 groups to receive 1) general anesthesia with propofol and remifentanil by continuous intravenous infusion at rates of 2 and 0.2-0.4 microgram/Kg/h, respectively; 2) subarachnoid anesthesia with lidocaine, and 3) subarachnoid anesthesia with 1.5% prilocaine. The second and third group received fixed doses of 3 mL of the local anesthetic. All patients were telephoned 48 hours after surgery and asked to answer 8 questions concerning prior experience of anesthesia, degree of satisfaction with the type of anesthesia used, postoperative pain, quality of information received about the anesthetic procedure, and undesirable side effects. RESULTS: We interviewed 120 patients and 32 refused to participate. All patients in the general anesthesia group would accept the same anesthetic technique again in future operations, whereas 85% and 82% in the lidocaine and prilocaine groups, respectively, would accept the same technique (p = 0.026). Satisfaction with anesthesia, postoperative pain, and quality of information about the anesthetic procedure was similar in all 3 groups. No important undesirable side affects were reported in any of the groups. CONCLUSIONS: The overall satisfaction with various types of anesthesia is similar among outpatients undergoing arthroscopic surgery on the knee; therefore, patient preferences for one technique over another should be taken into more consideration. PMID- 14753138 TI - [Premature mortality in the early postoperative period of liver transplantation]. AB - OBJECTIVES: To analyze the relation between premature death and complications that develop in the early postoperative period after liver transplantation. MATERIAL AND METHODS: Liver transplants performed in adults in our hospital were reviewed retrospectively. Patients were distributed in 2 groups, those who died and those who survived the early recovery period, and the incidences of complications in the two groups were compared using a chi 2 for single variables and logistic regression for multiple variables. RESULTS: In the 827 liver transplants studied, the recipient died in 86 cases, 7 inside the operating room (0.85%) and 79 during recovery (9.63%), for an overall mortality rate of 10.4% for surgery and the immediate postoperative period. Complications that were significantly different with respect to mortality in the multivariant analysis were post-reperfusion syndrome, atrial fibrillation, acute renal failure, and infection. CONCLUSIONS: Only one of the complications was specific to liver transplantation: post-reperfusion syndrome. The others were of the type found in any critically ill patient, an nearly all led to an increase in infections, which is the most common reason for death during surgical recovery. PMID- 14753139 TI - [Spanish scientific production in anesthesiology and resuscitation 1983-1995]. AB - OBJECTIVE: To quantify and analyze the information available internationally on Spanish scientific productivity in anesthesiology from 1983 through 1995. MATERIAL: Publications by Spanish authors indexed in three electronic databases: the Spanish Indice Medico (SIM), MedLine, and Embase (belonging to Excerpta Medica). METHOD: A bibliometric study of productivity as reflected by entries in the 3 databases, quantifying documents in circulation, change over time in circulating documents, overlap, and article's characteristics. RESULTS: During the period studied, a total of 2608 papers by Spanish anesthesiologists were indexed in the 3 databases (SIM 79%, Medline 54%, Excerpta Medica 29.5%). Annual growth was 16.7%, with a period of 10.2 years until duplication. CONCLUSION: Growth is exponential in anesthesiology, as in other disciplines, and the distribution satisfies Price's law. The 3 databases are the major collectors of the papers of Spanish anesthesiologists. The SIM and MedLine are the main indexers of Spanish-authored articles in the literature. Excerpta presents the largest growth on the compilation of articles. The inclusion of the three databases is essential in bibliographic searches and bibliometric studies on Spanish anesthesiology. PMID- 14753140 TI - [Intraoperative monitoring of the facial nerve: anesthesia and neurophysiology considerations]. AB - Intraoperative monitoring of the motor component of the 7th cranial pair or any other nerve should be routine during any surgical procedure involving risk of neural damage, whether or not the skull is opened. This paper discusses the main indications for monitoring facial and acoustic nerves during ear and neurosurgery involving the pontocerebellar angle (for acoustic neuroma or in surgery on the 8th cranial pair). Intraoperative electromyographic monitoring of the facial nerve should be used routinely in acoustic neuroma surgery to reduce the degree of postoperative neurological impairment and avoid possible malpractice suits. Although such monitoring requires that the muscle remain unblocked, intraoperative neuromuscular relaxants can be used if doses are administered along with appropriate monitoring of the level of peripheral neuromuscular block. PMID- 14753141 TI - [Inferior vena cava thrombosis after corrective surgery for tricuspid atresia (Fontan technique) in a pediatric patient]. AB - A 13-year-old boy weighing 35 Kg underwent surgery to correct pulmonary valve atresia by total caval-pulmonary anastomosis (Fontan procedure). During surgery and recovery, no adverse events were observed. On the third day after surgery, the patient developed signs of low cardiac output, renal failure, and right ventricular insufficiency. In spite of an antithrombotic treatment protocol with sodium heparin (10 IU/Kg/h), venous return was found to be impeded by 2 thrombi in the inferior vena cava. Thrombolysis with urokinase was ineffective, and surgical thrombectomy was only partially successful. Finally, a high dose of intravenous heparin was prescribed (25 IU/Kg/h) and the thrombi were dissolved. The importance of prophylactic antithromboembolic therapy following corrective surgery for congenital heart disease must be emphasized. Ultrasound monitoring of these patients, even in the absence of clinical signs, has proven effective for early detection of the thrombotic events described in this report. PMID- 14753142 TI - [Hypertriglyceridemic pancreatitis and pregnancy]. AB - A 33-year-old secundipara with a history of gestational diabetes and familial hypertriglyceridemia exacerbated during her previous pregnancy was admitted in the 36th week of gestation with diffuse abdominal pain, vomiting, low-grade fever, and general malaise. A blood sample had a lipemic, milky-pink appearance and plasma concentrations were as follows: triglycerides 2173 mg/dL, cholesterol 320 mg/dL, amylase 801 U/L, lactate dehydrogenase 650 U/L, creatinine 1.5 mg/dL, glucose 380 mg/dL, and left-shifted white cells. Acute pancreatitis was diagnosed and owing to signs of fetal distress, a cesarean was performed under light general anesthesia with propofol, succinylcholine, and sevoflurane. After the umbilical cord was cut, rocoronium and fentanyl were administered. The neonate was healthy and the patient's condition evolved favorably with conservative treatment. The incidence of pancreatitis during pregnancy is low but related morbidity and mortality are high. The usual cause is biliary tract disease, although rare metabolic alterations such as hyperlipidemia may occasionally act as the trigger. Early diagnosis and treatment are the keys to successful surgery and postoperative recovery. PMID- 14753143 TI - [Spinal and cerebral hematoma following diagnostic lumbar puncture with fatal course]. AB - A 64-year-old man suspected of having neurosyphillis suffered subarachnoid spinal and cerebral ventricular hemorrhage after a diagnostic lumbar puncture. The main risk factor was trauma occurring during a difficult puncture. Both the diagnosis and the subsequent laminectomy were delayed. Blood migrated to both lateral ventricles, complicating the clinical course, which ended in death. The need for rapid diagnosis and treatment in such cases is revealed by both images and clinical course. A pathophysiologic explanation for these hemorrhagic events is provided. PMID- 14753144 TI - [Lesion of the palatal mucosa: infrequent complication of nasotracheal intubation]. PMID- 14753145 TI - [Spontaneous pneumomediastinum (Hamman's syndrome). Anesthesia considerations]. PMID- 14753146 TI - [Assessment of recombinant human erythropoietin in scheduled orthopedic surgery]. PMID- 14753147 TI - [Pulmonary embolism caused by cement following vertebroplasty]. PMID- 14753148 TI - [Tattoos along the lumbar median line and epidural anesthesia]. PMID- 14753150 TI - [Innovative implants -- a prerequisite for biological osteosynthesis]. AB - Biological osteosynthesis is a new paradigm in fracture fixation. In contrast to the conservative mechanical operative period, where exact anatomical and rigid fracture fixation was the ultimate ambition, the new perception in fracture management attaches more importance to the conservation of bone perfusion than to the exact anatomical reduction. Because soft tissue condition and local blood circulation are most important for bone reconstruction and healing of fractures, additional damage of soft tissue caused by operation should ideally be avoided. Consequently surgeons aim at minimally invasive operating techniques using no touch bone implants. This shift in point of view has also been accommodated by a rapid progress in development of innovative bone implants. Solid medullar nails, novel designs and applications of plates, locking screws and biocompatible titanium implants are some of the implantation techniques which pushed the new concept of biological fracture fixation. PMID- 14753149 TI - [Are the terms "biology" and "osteosynthesis" contradictory?]. AB - Biology and Osteosynthesis may, at first glance, appear to be contradictory. The disadvantages of the surgical procedure are offset by improved recovery of blood supply across a fracture zone and by stimulation of callus formation due to (controlled and slight) instability (elastically flexible fixation). Biological internal fixation is a principle in its own right; its technique is to some extent incompatible with conventional compression fixation. Locked screws provide better anchorage of the implant in both biological and compression internal fixation techniques. This generally offers improved potential in the treating fractures in osteoporotic bone. Because locked screws offer improved anchorage, the use of unicortical screws is made possible with the effect that the medullary circulation is damaged less. Locked unicortical screws, especially if they are self-drilling and self-cutting, offer important surgical advantages in minimally invasive percutaneous osteosynthesis (MIPO) whereas bicortical screws are required for epi- and metaphyseal anchorage. PMID- 14753151 TI - [Biological internal fixation -- guidelines for the rehabilitation]. AB - The process of fracture healing restores the biological and mechanical state of the bone tissue. In contrast to other tissues, bone has the unique capacity to heal through a real repair process resulting not in a scar but in a regular reconstitution of its original tissue structure. During the last two decades, the understanding of bone biology continuously evolved leading to the new concept that preservation of the viability of the bone fragments is the key to unimpaired fracture healing. The biological fracture management provides environmental conditions that allow the natural healing process to occur as quickly and undisturbed as possible. In comminuted diaphyseal and metaphyseal fractures, the reduction process strives for restoration of proper length and axial and torsional alignment. Thereby, the reduction technique is mainly indirect, i.e. without direct visualization of the fracture area. The primary stability of an osteosynthesis seems to be of secondary importance for achieving sound bone healing. Stabilization can either be performed with an intramedullary nail or a bridging plate. The new biological plating technique imitates the concept of intramedullary fixation. It requires the use of a long implant working with improved leverage. This reduces both, the loading of the screws and the loading of the plate. The good healing capacity of viable fragments and their integration into the fracture callus protects the implant (especially a plate) from fatigue failure. To help to accomplish this new concept of biological plate fixation, new implants such as the Locking Compression Plate (LCP) have been introduced. The mechanical efficiency of this new plate generation is enhanced due to its possibility for angular stability screw insertion; load transfer by friction can be replaced by a load transfer by interlocking (internal fixator), reducing the biological interference of this implant (no contact implant). Functional rehabilitation with painfree mobilization needs to be performed carefully because the loading capacity of the biological plate osteosynthesis is low until radiological signs of bone healing via callus formation is visible. Clinical signs of overload and radiological indicators of potential mechanical failure of the fixation have to be identified by the general practitioners, and the compliance of the patient has to be controlled during the rehabilitation phase to avoid complications after biological internal fixation. PMID- 14753152 TI - [New concepts for stabilizing proximal humeral fractures]. AB - The treatment of proximal humerus fractures is overshadowed by problems like avascular humeral head necrosis, implant failure in osteopenic bone and moderate outcome, especially in the elderly. More precise imaging, a better understanding of the vascularity of the humeral head leading to indirect reduction techniques and new implants which allow for saver fixation even in osteopenic bone, allowing early functional aftercare, are the new promising factors which have the potential to ameliorate fracture treatment of the humeral head. A prerequisit for sucessful treatment is a thoroughful imaging which allows adequate preoperative planning. The type of stabilisation depends more on the surgeons preference and there are pros and cons with all implants. Functional after treatment however depends on the applied stabilisation principle and should be carefully monitored and adapted according to the bone healing process, monitored by regular postoperative x-ray controls. PMID- 14753154 TI - [Relevance and advantages of new angular stable screw-plate systems for diaphyseal fractures (locking compression plate versus intramedullary nail]. AB - Intramedullary nailing still remains the golden standard for the treatment of diaphyseal fractures of the long bones in adults. The operative technique is standardised and usually minimal invasive, the primary healing rate is high, and complications arise rarely. However, for problem zones (meta-diaphyseal region), in poor bone quality and in other relative contraindications for nailing (narrow medullary canal, fractures in adolescents and polytrauma) the new angular stable screw-plate-systems (LISS, LCP) offer an excellent alternative for the operative stabilisation. The high primary stability in combination with newly developed minimal-invasive techniques (MIPO = minimal invasive plate osteosynthesis) are the bases for a functional after treatment and a rapid bony consolidation with a low complication rate. PMID- 14753153 TI - [New principles in the surgical treatment of distal radius fractures -- locking implants]. AB - The treatment goal for fractures of the distal end of the radius is full functional recovery of the wrist. Prerequisites are restoration of the anatomy and early functional after treatment in accordance to the treatment guidelines for other articular and juxta-articular fractures. A better understanding of the anatomy and new biomechanical models lead to the development of new implants and operative techniques. The LCP-concept with locking screws had a considerable impact on recent advances in the operative treatment of distal radius fractures. PMID- 14753155 TI - [New minimally invasive methods of stabilizing distal femoral fractures]. AB - New, minimally invasive methods of stabilizing distal femur fractures offer the advantage of superior bone healing rates and lower infection rates despite of a decreased necessity of bone grafts. In comparison to conventional plating, the employed indirect reduction method does not lead to an increase in postoperative malalignments. The Less Invasive Stabilizing System (LISS) and the Distal Femur Nail (DFN) are introduced as examples of these new methods of stabilization. Although bone biology is widely respected by these new implants and operation techniques, distal femur fractures remain a serious injury, often resulting in subnormal function. PMID- 14753156 TI - [New stabilization techniques for fixation of proximal tibial fractures (LISS/LCP)]. AB - In unstable proximal tibia fractures secondary dislocation due to insufficient buttressing is a well known complication leading to incongruency of the joint and axial deviation of the leg. Double plate osteosynthesis increases stability, however it is not very biological and may be accomplished by wound healing problems. New stabilization techniques such as LISS (Less Invasive Stabilization System) and LCP (Locked Compression Plate) provide angle stability and minimal invasiveness. Therefore stability of fixation is improved and wound-healing problems decrease. Today simple fractures are still fixed with conventional plates and screws, however more complex bicondylar fractures, particularly those with a metaphyseal comminution zone and/or severe soft tissue damage are fixed favorably by the new stabilization techniques. PMID- 14753157 TI - [New surgical treatment procedures in fractures of the distal tibia (LCP, MIPO)]. AB - All along the operative treatment of distal tibial fractures is a difficult procedure. In most cases the clinical situation is characterized by small distal fragments in combination with crucial soft-tissue conditions. That's why complications as primary or secondary displacements, mal unions, delayed or non unions and as well as a high rate of deep wound infection are often seen. Thus internal fixations with traditional implants (standard screws and plates) could consider inevitable this crucial biology and biomechanics only insufficiently. The nowadays available internal fixators with optional angular-stable screws expand the possibilities of internal fixation in these severe situations. Their minimal invasive application (MIPO, Minimally Invasive Plate Osteosynthesis) takes care of the soft tissue and reduces the surgical trauma furthermore. With the variety of their possible applications (combination of angular stability with standard application) also the demands increase, however, both onto the surgeons, but also onto the general practitioners in the aftercare. The combination of most different tactics in one implant results in the consequence, that at the same bone simultaneously direct and indirect bone healing will be expected. The radiological differentiation between desired and unwanted healing processes becomes thus difficult. Pre- and perioperative procedures require from the trauma surgeon a huge infrastructure and a high measure of biomechanical and biological experience. In the postoperative management of these injuries an unlimited cooperation between traumatologists and general practitioners is indispensable for a further successful course. PMID- 14753158 TI - [Biological osteosynthesis in children and adolescents]. AB - Minimally invasive osteosynthesis techniques are very suitable for treatment of unstable fractures in children. Because of little additional soft tissue trauma, these techniques are also called biological osteosynthesis. They allow fracture treatment in children and adolescents without any harm to the growth plate. Angular deformities can be corrected due to further growth. Kirschner wires, threaded or unthreaded, elastic intramedullary rods or external fixators are most commonly used for osteosynthesis. After successful closed reduction of the fragments under x-ray amplifier control, the implants are inserted with minimal trauma to the soft tissues. This results in safe and rapid healing, spontaneous correction of slight malunions and fast restoration of function. The main disadvantage of these methods is a greater exposure of the patient and in particular the surgical team to x-ray radiation. These techniques result in shorter hospital stays enabling a fast return to a familiar environment. However high standard technical equipment and an experienced treating team are required for success. PMID- 14753159 TI - [Open reduction and internal fixation of a displaced transverse fracture of the sacrum with a locking compression plate]. AB - Isolated fractures of the distal sacrum are usually caused by direct trauma. Undisplaced fractures are easily missed on conventional X-ray films. CT scans can disclose the full extent of the fracture. Undisplaced, or minimally displaced fractures can be treated conservatively with analgesics and a restriction to seat for six weeks. In cases of major displacement or neurological symptoms, a digital transanal reduction of the fracture under local or regional anaesthesia can be attempted. If this manoeuvre fails, open reduction and internal fixation are required. Stabilisation is challenging because of the thin bone and the complex shape of the sacrum. In a 15 year old girl with major horizontal displacement and comminution of S4, following a snowboard accident, we achieved sufficient and stable fixation with the new LCP (Locking Compression Plate) system. For six weeks after the operation, the patient was not allowed to sit down. Thereafter, she was free of pain and unrestricted in daily activities and sport. The LCP system has a number of advantages in the fixation of fractures in osteoporotic or thin bone, but clinical experience/skill and careful planning of the operation are crucial. PMID- 14753160 TI - The European Astronaut Centre prepares for International Space Station operations. AB - The European Space Agency (ESA) contribution to the International Space Station (ISS) goes much beyond the delivery of hardware like the Columbus Laboratory, its payloads and the Automated Transfer Vehicles. ESA Astronauts will be members of the ISS crew. ESA, according to its commitments as ISS international partner, will be responsible to provide training on its elements and payloads to all ISS crewmembers and medical support for ESA astronauts. The European Astronaut Centre (EAC) in Cologne has developed over more than a decade into the centre of expertise for manned space activities within ESA by contributing to a number of important co-operative spaceflight missions. This role will be significantly extended for ISS manned operations. Apart from its support to ESA astronauts and their onboard operations, EAC will have a key role in training all ISS astronauts on ESA elements and payloads. The medical support of ISS crew, in particular of ESA astronauts has already started. This paper provides an overview on status and further plans in building up this homebase function for ESA astronauts and on the preparation towards Training Readiness for ISS crew training at EAC, Cologne. Copyright 2001 by the European Space Agency. Published by the American Institute of Aeronautics and Astronautics, Inc., with permission. Released to IAF/IAA/AIAA to publish in all forms. PMID- 14753161 TI - Advances in the diagnosis and treatment of dementia with Lewy bodies. Introduction. PMID- 14753162 TI - [Meta-analysis of GEMZAR platinum in non-small cell lung cancer]. PMID- 14753163 TI - [Abstracts from the VIth European Congress of Endocrinology. Lyon, France, April 2003]. PMID- 14753164 TI - Beyond antimicrobials--the future of gut microbiology. Abstracts of the 3rd Joint RRI-INRA Gastrointestinal Tract Microbiology Symposium. 12-15 June 2002, Aberdeen, Scotland, United Kingdom. PMID- 14753165 TI - [Potential and limitations of fluorescence diagnosis and photodynamic therapy. Part I: Fluorescence diagnosis]. AB - Fluorescence diagnostic methods enable tumor visualization via specific excitation of endogenous or exogenous tumor-selective fluorescent markers and subsequent optical measurement of the induced fluorescence signals. In the following review, the reader will be given a clear and extensive understanding of all aspects of these methods relevant for ENT specialists. Apart from a demonstration of the clinical difficulties in the diagnosis of malignant tumors of the upper aerodigestive tract, a historical synopsis of fluorescence diagnosis and an explanation of the biophysical basics, this publication focuses mainly on a detailed discussion of the current status of research and a critical consideration of the commercially available systems for fluorescence diagnosis in ENT oncology. The topic is rounded off by a short account of possible future activities in this field. PMID- 14753166 TI - [Work shops]. PMID- 14753167 TI - Pharmacovigilance in Clinical Practice. Abstracts of the ISoP Annual Conference. Marrakech, Morocco, 8-11 October 2003. PMID- 14753168 TI - Rectal surgery for endometriosis--should we be aggressive? PMID- 14753169 TI - Organ Microcirculation: A Gateway to Diagnostic and Therapeutic Interventions. Abstracts of the 14th Keio University International Symposium for Life Sciences and Medicine. December 11-13, 2003, Tokyo, Japan. PMID- 14753170 TI - [Hessian Natural Medicine Award 2003]. PMID- 14753171 TI - [Guidelines for advanced life support in cardiology--code blue--resuscitation registry. Standardization of the emergency cart]. PMID- 14753172 TI - [Good fortune and eyeglasses]. PMID- 14753173 TI - [Goal-oriented marketing for general practice]. PMID- 14753175 TI - Abstracts of the Pakistan Society of Clinical Oncology 10th Annual Cancer Conference. 19-21 December 2003, Peshawar, Pakistan. PMID- 14753174 TI - [Health care reform 2003. Consequences for the contract physician]. PMID- 14753176 TI - Australian Dental Research Foundation research grant abstracts. PMID- 14753178 TI - [XXII Congress of the European Academy of Allergology and Clinical Immunology]. PMID- 14753177 TI - Physiological and proteomic approaches to studying prefertilization events in the human. AB - This research aims firstly to understand, in cellular and molecular terms, how a mature human spermatozoon is prepared for fertilization, and secondly, to identify what factors are involved in the initial signalling interactions between the egg and spermatozoon. In order to achieve these objectives, a combination of approaches is being used, including single-cell imaging, patch clamping and proteomics. Single-cell imaging reveals hidden complexity and heterogeneity in signalling responses in spermatozoa. Characterization of cell physiology at the single-cell level must be a future aim, including the study of ion channel expression and function by patch clamping. Proteomic experiments are aimed at identifying defects in protein expression in specific subgroups of men, e.g. those with globozoospermia. A better understanding of prefertilization events will allow the development of non-assisted reproductive therapy, drug-based treatments for male infertility. PMID- 14753179 TI - [Medications in orthopedics. Data, facts, evidence-based statements]. PMID- 14753180 TI - Cumulative index volumes 26-47. PMID- 14753181 TI - Insulin resistance syndrome. PMID- 14753183 TI - [Intraoperative radiotherapy as part of the treatment concept of soft tissue sarcomas]. AB - The therapy of soft tissue sarcomas is not uniform. The goal of treatment for soft tissue sarcomas is surgical resection with negative microscopic margins. Other therapy options are pre-, intra-, or postoperative radiotherapy as well as neoadjuvantor adjuvant chemotherapy with different drugs, systemic or under local perfusion of the limb. If a margin-free surgical resection is not possible, intraoperative radiation therapy (IORT) offers the possibility for treatment of sites at high risk for local recurrence. The goal of this collaborating study of the Orthopedic University Hospital of Heidelberg and the department of Radio Oncology of the University Hospital of Heidelberg is to observe the influence of IORT on the rate of local recurrence, complications, metastases, and survival in another wise unchanged tumor management. From 1960 to 1998,78 patients were treated for soft tissue sarcomas; 26 of 78 patients underwent intraoperative radiation and 52 patients were treated without intraoperative radiation. We found a recurrence rate of 15% in the IORT group and 29% in the non-IORT group. No difference for survival or distant recurrence was found, also no difference in early postoperative complication rate, but a higher rate of late complications after IORT (34% vs 7%) was remarkable. We think intraoperative radiation allows better local control and for that reason IORT is integrated into our therapeutic concept. Its influence on survival and distant metastases needs further investigations. PMID- 14753184 TI - Review of position paper on community-based education. PMID- 14753185 TI - [Obstructive sleep-apnea syndrome and narcolepsy]. PMID- 14753186 TI - [Abstracts of the 31st Annual Meeting of the Austrian Society of Diabetes. 27-29 November 2003]. PMID- 14753188 TI - Tennessee Dental Association directory and membership roster 2003-2004. PMID- 14753187 TI - [Septic loosening of a Wagner revision stem provoked by Mycobacterium tuberculosis]. AB - Only a few cases of specific tuberculous infections of total hip arthroplasties have been published. We report the case of a 66-year-old male patient who received a cementless total hip arthroplasty due to osteoarthritis of the left hip in 1990. Four years later, revision arthroplasty with a Wagner revision stem was performed because of aseptic loosening. In 1995 revision of a loosened acetabular cup was necessary. In 1996 we saw the patient for the first time in our outpatient unit. He complained of increasing pain in the region of the left hip. X-rays showed loosening of the Wagner stem. Aspiration of the synovial fluid of the left hip revealed an infection with mycobacterium tuberculosis (radioimmunoassay). There were no signs of tuberculous infection in the patient's history. Treatment consisted of removal of the prosthesis followed by antituberculosis chemotherapy for 12 months with rifampicin, ethambutol, isoniazid, and pyrazinamide. In April 2001 revision of the left hip joint and implantation of a MRP titanium revision stem and reconstruction of the acetabulum with an acetabular reconstruction ring was done. Until today the patient exhibits no signs of recurrence of the tuberculous infection. PMID- 14753189 TI - [Magnetic resonance imaging spectroscopy. Part 1: Basics]. AB - A century after the discovery of X-rays, the low-energy range of the electromagnetic spectrum also attained broad application in radiology. Radiofrequency waves allow excitation in a magnetic field of the magnetic resonance of spin-bearing nuclei in tissue. Using the intense signal of the water protons, morphological images of the human body can be obtained, while at a higher frequency resolution also endogenous metabolites as well as pharmaceuticals, which contain MR-visible nuclei (e.g., 1H, 13C, 19F, 31P), can be detected noninvasively and in vivo. Accordingly, in vivo MR spectroscopy is a technique which is sensitive to molecules and molecular properties and which can be applied to repeated examinations. Its major limitation is the low signal intensity vs noise, which implies long measurement times and poor spatial resolution. Using spectroscopic imaging, the distribution of metabolites within an organ can be monitored selectively and displayed as a molecular image. PMID- 14753190 TI - [The pediatric chest--new possibilities in the radiological evaluation]. PMID- 14753191 TI - [A law to limit embryo transfer in ART?]. PMID- 14753192 TI - Oral health policies. American Academy of Pediatric Dentistry. PMID- 14753193 TI - Clinical guidelines. American Academy of Pediatric Dentistry. PMID- 14753194 TI - Mental Health Policy and Economics--The Value of Research. Abstracts of the 6th Workshop on Costs and Assessment in Psychiatry. Venice, Italy, March 28-30, 2003. PMID- 14753195 TI - [The G-DRG system 2003/2004 from the trauma surgery-orthopedic viewpoint. A strengths and weaknesses analysis]. PMID- 14753196 TI - [Diagnostics at the wrist]. PMID- 14753197 TI - Proceedings of the 47th Congress of the Korean Society of Veterinary Science. Daejeon, Korea, October 17, 2003. PMID- 14753198 TI - Intestinal spirochaetes of the genus Brachyspira: an update on recent findings. PMID- 14753199 TI - [Endoprosthetic replacement of the ankle joint]. AB - Endoprosthetic replacement of the ankle joint is considered to be a modern alternative of the well-tried fusions of the joint. We try to explain indications and limits of alloarthroplasty in comparison to arthrodesis. The recent technical evolution will be presented: starting with the two-component-("first generation") and leading to the three-component ("second generation") designs, Suitable for cemented and cementless implantation as well. Results of three-component endoprostheses in the literature and our own experiences with implants of both generations especially regarding the time of survival will be discussed. Basing on the actual knowledge, we try to deduce a prognosis of the future way of ankle replacement. PMID- 14753200 TI - A model of magnesium metabolism in young sheep: transactions between plasma, cerebrospinal fluid and bone. AB - An extension of the previously proposed model of Mg metabolism (Robson et al. 1997) has been developed to consider the transactions of Mg that are associated with cerebrospinal fluid (CSF) and bone. The representation of the CSF as a single MG compartment with uptake from the plasma described by Michaelis-Menten kinetics gives very good agreement with published experiments. Analysis of the available information on resorption of Mg from adult bone indicated that this process makes a negligible contribution to Mg homeostasis and can be omitted from the model. PMID- 14753201 TI - Assessment and comparison of intraoperative and postoperative pain associated with NovaSure and ThermaChoice endometrial ablation systems. PMID- 14753202 TI - Selection of abstracts submitted for the AAGL annual meeting: an unbiased process. PMID- 14753203 TI - One century of liquid chromatography. From Tswett's columns to modern high speed and high performance separations. PMID- 14753204 TI - Frequently relapsing nephrotic syndrome: treatment with mycophenolate mofetil. AB - Long-term treatment with cyclosporin A (CyA) of children with frequently relapsing steroid-sensitive nephrotic syndrome (SSNS) carries the risk of nephrotoxicity. We have analyzed renal function in 23 children with SSNS during CyA therapy. Repeated measurements of glomerular filtration rate (single-shot 51Cr-EDTA clearance) showed a decline from 131 +/- 21 ml/min per 1.73 m2 to 116 +/- 27 ml/min per 1.73 m2 at last follow-up. Similarly, effective renal plasma flow (simultaneous 123 I-hippurate clearance) was correlated with duration of CyA treatment, and showed a decline from 980 +/- 318 ml/min per 1.73 m2 to 724 +/- 242 ml/min per 1.73 m2. In a pilot study we investigated the effect of mycophenolate mofetil (MMF) in 7 children with a median age of 12.7 years [6 with minimal change nephrotic syndrome (MCNS), 1 with focal segmental glomerulosclerosis (FSGS)] with signs of nephrotoxicity because of long-term CyA therapy. Only 1 patient with SSNS showed a relapse during MMF therapy. In the patient with FSGS, MMF was started in addition to CyA, resulting in complete remission for a follow-up of 28 months. This preliminary study demonstrates that children with MCNS treated with CyA may be successfully converted to MMF without major side effects. In all cases, including FSGS, MMF had a beneficial effect on renal function. These data should be confirmed by a prospective randomized clinical trial. PMID- 14753205 TI - UN urges broadcasters to air AIDS programmes. Entertainment is better than factual films for increasing awareness. PMID- 14753206 TI - Shopping 'til we drop. Can psychology save us from our lust for possessions? PMID- 14753207 TI - Six babies die in Russia from hospital-acquired infection. PMID- 14753208 TI - Australian drugs scheme threatens free-trade talks. PMID- 14753209 TI - Thai HIV vaccine trial prompts angry exchanges. PMID- 14753210 TI - IOM calls for universal health coverage by 2010. PMID- 14753211 TI - Suicide rates rise among soldiers in Iraq. PMID- 14753212 TI - HIV/TB plan expanded. PMID- 14753213 TI - Cost deal for HIV tests. PMID- 14753214 TI - Blood transfusion scare. PMID- 14753216 TI - Drive to improve patient choice within the UK NHS. More patient involvement in treatment decisions could lead to more efficient NHS, experts say. PMID- 14753215 TI - Helicobacter pylori eradication down-regulates matrix metalloproteinase-9 expression in chronic gastritis and gastric ulcer. PMID- 14753217 TI - The true ligand of the NOD2 receptor is peptidoglycan instead of lipopolysaccharide: a schematic representation of ligand-receptor interactions and NF-kappa B activation. PMID- 14753218 TI - Chronic hepatitis C after liver transplantation: a randomized study. PMID- 14753219 TI - Pitfalls of pit pattern diagnosis in ulcerative colitis-associated dysplasia. PMID- 14753220 TI - Further validation of high-magnification chromoscopic-colonoscopy for the detection of intraepithelial neoplasia and colon cancer in ulcerative colitis. PMID- 14753221 TI - Prevention is the best defense: surgical prophylaxis of pouchitis. PMID- 14753222 TI - Limitations of transjugular intrahepatic portosystemic shunt for management of gastric varices. PMID- 14753223 TI - Discussion on antro-fundic dysfunction in functional dyspepsia. PMID- 14753224 TI - Increased risk of noncardia gastric cancer associated with proinflammatory cytokine gene polymorphisms. PMID- 14753225 TI - Discussion on impaired expression of peroxisome proliferator-activated receptor beta in ulcerative colitis. PMID- 14753226 TI - [Cutting edge medicine and medical ethics: medical ethics in research-type clinical studies and translational research coordinators (TRC)]. PMID- 14753227 TI - [Evidence in otorhinolaryngology and head and neck surgery--evidence in the treatment of early laryngeal (glottic type) cancer]. PMID- 14753228 TI - [Respiratory Rehabilitation Manual--physical therapy]. PMID- 14753229 TI - [WMN: a negative ERPs component related to working memory during non-target visual stimuli processing]. AB - OBJECTIVE: To study non-target stimuli processing in the brain. METHOD: Features of the event-related potentials (ERPs) from non-target stimuli during selective response task (SR) was compared with that during visual selective discrimination (DR) task in 26 normal subjects. The stimuli consisted of two color LED flashes (red and green) appeared randomly in left (LVF) or right (RVF) visual field with same probability. ERPs were derived at 9 electrode sites on the scalp under 2 task conditions: a) SR, making switch response to the target (NT) stimuli from LVF or RVF in one direction and making no response to the non-target (NT) ones; b) DR, making switching response to T stimuli differentially, i.e., to the left for T from LVF and to the right for T from RVF. RESULT: 1) the non-target stimuli in DR conditions, compared with that in SR condition, elicited smaller P2 and P3 components and larger N2 component at the frontal brain areas; 2) a significant negative component, named as WMN (working memory negativity), appeared in the non target ERPs during DR in the period of 100 to 700 ms post stimulation which was predominant at the frontal brain areas. CONCLUSION: According to the major difference between brain activities for non-target stimuli during SR and DR, the predominant appearance of WMN at the frontal brain areas demonstrated that the non-target stimulus processing was an active process and was related to working memory, i.e., the temporary elimination and the retrieval of the response mode which was stored in working memory. PMID- 14753230 TI - [Effects of repeated body position change training for human orthostatic tolerance]. AB - OBJECTIVE: To verify the validity that repeated position change training can increase human orthostatic tolerance and to explore its mechanism. METHOD: Six subjects were trained with repeated position change for 11 d according to a protocol of alternative head-down and head-up tilts, each set of training lasted for about 35 min. Their orthostatic tolerances were determined before and after training. RESULT: Compared with the data before training, subjects' symptom scores during orthostatic tolerance test after training decreased significantly (4.50 +/- 1.05 vs. 2.83 +/- 1.60, P<0.05), magnitude of the increased heart rate increase lowered significantly [(29.3 +/- 4.3) bpm vs. (13.5 +/- 7.5) bpm, P<0.01], magnitude of mean arterial blood pressure augmentation increased significantly [(4.8 +/- 4.4) mmHg vs. (9.0 +/- 3.0) mmHg, P<0.05] and cardiovascular response index decreased significantly (34.42 +/- 5.00 vs. 22.33 +/- 8.27, P<0.01). In brief, the responses to orthostatic stress were improved after training. CONCLUSION: Repeated alternative body position change training can increase human orthostatic tolerance. This kind of training is promising for pilot, especially astronaut training. PMID- 14753231 TI - [Effects of short-action hypnotics triazolam and zopiclone on simulated flight performance]. AB - OBJECTIVE: To evaluate the effects of short-action hypnotics triazolam and zopiclone on simulated flight performance so as to provide experimental evidences for the use of these drugs in aircrew. METHOD: Six healthy young male volunteers served as the subjects. They were asked to take triazolam (0.25 mg), zopiclone (7.5 mg) or placebo at noon time (13:00) and sleep for 1 h. Simulated flight performances were tested in a simulator 1 h, 2 h, 3 h, 4 h, 6 b, 8 h, 10 h and 20 h (next morning 9:00) after taking the drugs. The experiments were done once a week for 3 weeks. RESULT: No significant difference was found between simulated flight performances before or after triazolam while simulated flight performances at 2 h and 3 h after taking zopiclone showed significant decrease (P<0.05), but restored to normal 4 h after taking the drug. CONCLUSION: Short-action hypnotics triazolam had no significant effect on simulated flight performance, while zopiclone had significant adverse effects on simulated flight performance at 2 h and 3 h after taking the drug. PMID- 14753232 TI - [Circadian rhythms of DNA synthesis and telomerase expression in hepatic cancer transplanted in nude mice]. AB - OBJECTIVE: To study the circadian rhythms of DNA synthesis and telomerase expression in hepatic cancer transplanted in nude mice. METHOD: Sixteen BALB/C mice were synchronized with an alternative lighting regimen with 12 h for light and 12 h for darkness (12:12 LD) for 4 weeks. Hepatic cancer cells (SMMC-7721) were implanted into both flanks of each mouse. One week after transplantation, sampling from the tumor was conducted at 3, 9, 15 and 21 h after light onset (HALO). Single cell suspension was obtained and stained with propidium iodide. The cellular DNA content was measured by flow cytometry. Telomerase activity was measured by PCR-ELISA assay. Data were documented by ANOVA and Cosinor analysis. RESULT: The proportion of tumor cells in phase G1, S, G2/M and telomerase activity varied according to circadian time with statistical significance, and the telomerase activity showed a synchronized variation. The distribution curves of both phase S and the expression level of telomerase were fit for Cosinor changes. CONCLUSION: DNA synthesis and telomerase expression of SMMC-7721 cells transplanted into the nude mice varies according to the circadian rhythm. The results provide a guidance for laying down the chemotherapy protocol for human tumors, especially when the telomerase inhibitor was used as the anti-cancer agent. PMID- 14753233 TI - [Numerical analysis of finger temperature distribution in a cold environment]. AB - OBJECTIVE: Temperature distribution over the finger was discussed, so as to provide reference for thermal protection of the hand. METHOD: Mathematical model for thermoregulation of trunk including hands was established. In the model, the trunk was divided into 15 segments and the hand was divided into palm and 5 fingers. The model was presented as a consistent set of bio-heat equations and was solved by a finite element method. RESULT: Effect of wind speed on the finger temperature is so evident, while the effect of artery blood temperature on the finger temperature is so little. CONCLUSION: Through the comparison with the result of literature, the establishment of the model was proved to be reasonable and the method feasible. PMID- 14753234 TI - [Aerodynamic characteristics of crewman's arms during windblast]. AB - OBJECTIVE: To study the aerodynamic characteristics of crewman's arms with or without protective devices in the status with raised legs or not. METHOD: The experiments were performed in an FL-24 transonic and supersonic wind tunnel, over Mach number range of 0.4-2.0, with 5 degrees-30 degrees angles of attack, 0 degrees - 90 degrees sideslip angles and Re number of (0.93-3.1) x 10(6). The test model was a 1/5-scale crewman/ejection seat combination. The aerodynamic characteristics of the various sections of crewman's arms were studied and analyzed. RESULT: The results showed that 1) The effect of raised leg on the aerodynamic characteristics of the crewman's arms was very evident, and was related to the status of leg raising; 2) The sideslip considerably increased aerodynamic loads on the crewman's arms, in particular when beta=50 degrees the loads was severe in the test; 3) The tested protective devices was valid, the effectiveness of wind deflector in protecting crewman's arms was evident; 4) A formula for calculating aerodynamic force acting on crewman's arms was presented. CONCLUSION: 1)The tested protective devices was valid, and the effectiveness of wind deflector in protecting crewman's arms was evident; 2) An aerodynamic basis for the development of crewman windblast protective device was presented; 3)The calculation formula presented is useful in estimating aerodynamic forces of crewman's arms. PMID- 14753235 TI - [Principles and experimental analysis of a measuring system for measuring spacesuit joint's damping parameters]. AB - OBJECTIVE: To measure the damping parameters of the spacesuit joint. METHOD: The principles of the passive robot system for measuring spacesuit joint's damping parameters were presented. Basing on its special mechanical structure, a 3 DOF model of the flexible IVA (intra-vehicular activity) spacesuit's sleeve was built. The optimal approximation of inverse calculation based on 6 dimension space interval was described. The damping parameters of an actual IVA spacesuit's sleeve were measured on the actual testbed. Also, the potential application of the measuring system was discussed. RESULT: The model of spacesuit sleeve and the forward/inverse kinematics were proved by experimental measurements and real time 3D simulation. CONCLUSION: The principles and the proposed method of the measurement were testified. PMID- 14753236 TI - [Decomposition of EMG signals based on combination of information diffusion theory and fuzzy neural network]. AB - OBJECTIVE: To solve the problem of large samples and contradictory samples in EMG during high level muscle contraction. METHOD: By means of recording EMG during muscle contraction with linearly increasing force instead of constant force, basic MUAP templates were obtained with the combination of information diffusion theory and fuzzy neural network. Samples were compressed and contradictory samples were eliminated. RESULT: The method was tested by simulated and real EMG data and the results were satisfactory. CONCLUSION: This method is meaningful for decomposing NEMG at high level muscle contraction. PMID- 14753237 TI - [Measurement and statistical analysis of the vertical force in normal young people's gait]. AB - OBJECTIVE: To find a method for evaluating the relationship between mechanical data and gait efficiently. METHOD: Gait mechanical data of 27 young men (20-22 years) without history of the kinetic diseases were collected using a force measuring system. The data were transformed into characteristic two-peak curves in vertical direction (z-axis). Seven characteristic parameters were distilled. The coefficient of variation (CV) and the absolute symmetry index (ASI) of the parameters between left foot and right foot were analyzed. RESULT: It was found that CV of the first temporal parameter of gait (T(Z1)) is equal to 14.49%, which corresponds to larger dispersion. The values of ASI are all lower than 12.5%, which means the symmetry between left and right feet meets general requirement. But the P value (result of t-test) of F(Z2) is 0.0368 (lower than 0.05), which means that there is obvious differentia between F(Z2) of left and right feet, while no such differentiae were found for the other six characteristic parameters. t-test was also used to analyze the differentia between the gait of male and female. It is still found that p value of F(Z2) is lower than 0.01, which corresponds to a great differentia on F(Z2) between male and female. CONCLUSION: Some significant conclusions of the repetition, dispersion, symmetry and differentia of gait data have been obtained by the analysis. PMID- 14753238 TI - [A new membrane-permeable peptide from human]. AB - OBJECTIVE: To study the translocation ability of a new protein domain (Circadian locomotor output cycles kaput protein's DNA-binding peptide, hCLOCK's DNA_BIND) from human through the membrane. METHOD: hCLOCK's DNA_BIND, which was chemically synthesized and labeled with FITC at N-terminal, was incubated with the vascular endothelial cell (ECV-304) and primary culture neuroglial cells in the culture medium, the distribution of it was observed on a fluorescence microscope. RESULT: hCLOCK's DNA_BIND has the ability to translocate through the cell membrane and accumulate in the nucleus. The quantification of cellular uptake of it increases with the increase of incubation time and concentration. However, the peptide was internalized at 4 degrees C as efficiently as that at 37 degrees C. CONCLUSION: It is promising to provide a new safe carrier for the intracellular treatment. PMID- 14753239 TI - [Measurement of cerebral blood oxygen saturation with near infrared spectroscopy and its application in hypergravity]. AB - OBJECTIVE: To measure the oxygen saturation of blood in rat's brain non invasively after acceleration with near infrared spectroscopy technique. METHOD: A serious of model about different ischemia and hypoxia of head, measured oxygen saturation by phlebotomize were made, and the difference with normal group was obtained. Its absorption intensity was measured and a curve showing the difference between the normal value was drawn. According to the curve, cerebral blood oxygen saturation after acceleration was calculated with the measured absorption intensity after high or push-pull maneuver by near infrared spectroscopy. RESULT: Compared high +Gz and push pull effect group with 0 Gz group, the content of ox hemoglobin reduced, in opposition the deox hemoglobin increased, the greater the difference, the lower the content of oxygen saturation of blood. CONCLUSION: Oxygen saturation of blood in the rat's brain can be obtained non-invasively with near infrared spectroscopy. PMID- 14753240 TI - [Effect of space flight on yield of Monascus purpureus]. AB - OBJECTIVE: To select high Lovastatin-producing microbial breed by space flight. METHOD: Monascus purpureus species was carried into space by the recoverable spaceship, "Shenzhou 3". After flight, the strain was rejuvenized, segregated and selected. The content of Lovastatin produced in the solid fermentation was examined. RESULT: Mutants with high productivity of Lovastatin were obtained. A series of tests showed that the acquired character of the mutants was stable. CONCLUSION: Space flight is an effective method for the selection of fine strains. PMID- 14753241 TI - [Effects of enriched oxygen on human free radical metabolism during exercise at 5 380 m plateau]. AB - OBJECTIVE: To study the effect of enriched oxygen on free radical metabolism at high altitude. METHOD: SOD, GSH-Px and MDA were measured in 10 health young men after step in a room with or without oxygen enrichment at high altitude of 5 380 m. RESULT: After exercise in the oxygen-enriched room, SOD and GSH-Px increased significantly while MDA decreased notably (P<0.01) than those after exercise in the room without oxygen enrichment (P<0.05 or P<0.01). CONCLUSION: Enriched oxygen can decrease damage from free radical at high altitude. PMID- 14753242 TI - [Effects of simulated weightlessness on reproduction in male rats]. AB - OBJECTIVE: To study the effects of tail-suspension on gonad and spermatic hormone in male rats. METHOD: After tail-suspension and reambulation some times in male rats, weight of testis was measured, sperm of epididymis was inspected, morphology of testicular cell was compared, and antent of testosterone assay by radioimmunoassay ratio of LH-beta positive cell in pituitary was measured by immunohistochemic. RESULT: After tail-suspension, weight of testis in male rats decreased significantly. Shape of the convoluted tubules and number of all levels of the spermatocyte in testis atrophied markedly. No sperm was found in the epididymis. Content of testosterone decreased markedly. Number of positive cell of beta-subunit of LH in the pituitary increased markedly. CONCLUSION: Tail suspension has negative effect on sex gland and reproduction function of male rats. PMID- 14753243 TI - [Study on space motion sickness before, during and after spaceflight]. AB - Space motion sickness has long been a difficult medical problem in human space flight. Because it is a group of syndromes of the astronauts in adaptation to the new space environment and is related to a wide range of factors, the research of space motion sickness is different from that of cardiovascular and pulmonary system. Subjective methods and qualitative investigation are frequently used, and quantitative analysis is limited since objective stimuli that can be used in specific space environment are few and the samples are small. So precise localization of the problem is very difficult. In this paper space vestibular experiments and works on perceptions of space motion sickness completed by Russian and American scientists are reviewed, in the hope that it may provide some references for future space medical researches. PMID- 14753244 TI - [sEMG Time-frequency analysis techniques for evaluation of muscle fatigue and it's application in ergonomic studies]. AB - As a non-invasive on-line measurement, sEMG can reflect the status of muscle activity and muscle function accurately and objectively. Some sEMG Time-frequency analysis techniques, especially the JASA (joint analysis of EMG spectrum and amplitude) analysis, for evaluation of muscle fatigue in ergonomics and occupational field studies are introduced and evaluated in this paper. The sEMG signal analysis and the necessity for developing sEMG analysis techniques for field use in ergonomics are also briefly discussed. PMID- 14753245 TI - Budesonide Modulite: improving the changeover to CFC-free treatments. PMID- 14753246 TI - The formulation and evaluation of a CFC-free budesonide pressurised metered dose inhaler. AB - Although dry powder inhalers are well established for the delivery of corticosteroids, the pressurised metered dose inhaler remains the preferred and most cost effective presentation. To design an HFA solution formulation which matched marketed CFC products (Pulmicort and Desonac DA) two elements of the Chiesi Modulite system, the addition of a non-volatile component and the actuator orifice diameter, were varied. These variables, which were shown by in vitro tests to influence the fine particle dose and its mean particle size in different ways, could be permuted to give an aerosol cloud with size characteristics very close to the comparator products. The likelihood that this would confer clinical equivalence is reinforced by a pharmacokinetic analysis which showed that the chosen HFA solution formula gave similar systemic absorption from the lung as Pulmicort. The equivalence in aerosol characteristics was sustained when the pressurised metered dose inhalers (pMDIs) were used with spacers. The Chiesi Jet and the AstraZeneca Nebuhaler, when used with their respective pMDIs, reduced likely oropharyngeal deposition to the same extent and gave a similar increase in the fine particle dose. PMID- 14753247 TI - Equivalent asthma control and systemic safety of inhaled budesonide delivered via HFA-134a or CFC propellant in a broad range of doses. AB - The aim of the present study was to demonstrate an equivalent asthma control and safety of inhaled budesonide 200 microg unit-dose via a spacer device (Jet Spacer, Chiesi Farmaceutici S.p.A.) given with an HFA-134a or CFC propellant in stable patients treated with inhaled corticosteroids. A total number of 270 patients, 134 in the HFA-134a group and 136 in the CFC group, completed a 2-week run-in period and were then randomised to receive a daily dose of inhaled budesonide (low dose: 400 microg, medium dose: 800 microg, high dose: 1200 or 1600 microg), defined on the basis of the dose of previous inhaled steroids given twice daily for 12 weeks. Morning and evening PEFR, intake of rescue salbutamol, number of day-time and night-time asthma attacks, number of night-time awakenings due to asthma and clinical symptoms were recorded daily by patients on diary cards. Pulmonary function tests (FEV1, FVC, PEFR and MEF50) and vital signs were measured at the clinics at study entry, at the start of treatment and after 2, 4, 8 and 12 weeks thereafter. Morning serum cortisol (8.00-10.00 AM) was measured at baseline and in the final visit. Adverse events and vital signs were recorded throughout the total study period. Small increases vs. baseline for lung function (more markedly in the high-dose subsets) and significant decreases of symptoms and use of rescue salbutamol were similarly observed in both groups. Equivalence was demonstrated for the primary endpoint morning PEFR (difference between means = -1.51 l/min; 95% CI: -9.40-6.37 l/min; pre-defined limits: +/- 42.16 l/min, i.e. +/- 10% of the reference LSM) as well as for evening PEFR and FEV1, both in the ITT population or on a per-protocol basis. No statistically significant differences between groups were observed in any of the other efficacy variables. A similar proportion of drug-related adverse events was observed in the two groups, without drug-related serious events in either group. No evidence of adrenal depletion was also noted with both propellants. In conclusion, the budesonide HFA-134a formulation given with a spacer device provided an equivalent asthma control with that of a corresponding CFC product, when administered in stable patients treated with inhaled corticosteroids in a broad range of daily doses. The use of the new propellant did not modify the safety profile of inhaled budesonide. PMID- 14753249 TI - Degradation of estrogen conjugates using titanium dioxide as a photocatalyst. AB - Estrogen conjugates (estradiol-3-glucuronide, -17-glucuronide, estrone glucuronide and -sulfate) were subjected to photodegradation using titanium dioxide immobilized on glass beads as a catalyst. Their time courses were measured by HPLC and compared with those of the unconjugated estrogens. Estradiol, its 17-glucuronide and estrone, which have an unconjugated phenolic hydroxy group at the C-3 position, were almost completely degraded by UV irradiation within 4 h. On the other hand, significant amounts of estradiol- and estrone-3-glucuronide (ca. 20%, 25%) and estrone sulfate (ca. 90%), which were conjugated at the 3-hydroxy group, remained after a 6.5 h irradiation. These results supported the hypothesis that the photodegradation of estrogens was initiated at the phenolic hydroxy group. PMID- 14753248 TI - Efficacy and safety of inhaled budesonide delivered once or twice daily via HFA 134a in mild to moderate persistent asthma in adult patients. Comparison with budesonide CFC. AB - This study was undertaken to investigate whether budesonide 4001 microg twice daily (Chiesi Farmaceutici S.p.A.) given with the HFA-134a propellant is equivalent in efficacy and safety to the same dose regimen delivered with the marketed CFC product in adult asthmatics with mild to moderate persistent asthma; the effects of budesonide HFA 800 microg once daily were also studied. After a 2 week run-in, a total number of 98, 103 and 97 patients were assigned to the 12 week treatment with budesonide given with HFA or CFC twice daily (morning and evening), or HFA once daily (morning), respectively. The main outcome variable morning PEFR, as well as evening PEFR and clinical symptoms (day-time and night time asthma attacks, number of asthma-induced night-time awakenings and overall symptoms' scores) were measured daily by patients. Other standard pulmonary function testing were measured at clinic visits. A blood sample for morning serum dosing (8.00-10.00 AM) was taken at baseline and at endpoint. Adverse events and vital signs were also recorded. Significant improvements at endpoint in morning and evening PEFR, as well as in clinic PEFR and MEF50, were observed in both the twice daily groups only. An exact proof of equivalence between HFA and CFC given twice daily was demonstrated for the primary parameters, morning PEFR (equivalence pre-defined limits were +/- 40.27 l/min, difference between means = 4.0 l/min and 95% CI -6.9-14.9) and secondary parameters as evening PEFR: (limits +/- 40.19 l/min, difference between means = 2.1 l/min and 95% Confidence interval (CI) -9.4-13.5) and FEV1 (limits +/- 0.27 l, difference between means = 0.0 l and 95% CI -0.11-0.10). Less evident (but within limits) proofs of equivalence were shown in the comparisons with the once daily group. No substantial differences between the three groups were observed for the other efficacy variables, including symptoms and use of rescue salbutamol, which significantly improved over the run-in values in all groups. Minimal and non-significant decreases over pre-treatment values were observed in the three groups for morning serum cortisol levels: the analysis of individual data has shown a better outcome in the HFA twice daily regimen, compared with the other two groups. Again, a similar amount of patients in both the twice daily groups reported drug-related adverse events, which were more frequent in the once daily HFA group. Therefore, the results of this study have shown that inhaled budesonide given with new HFA-134a propellant can replace microgram-equivalent doses of the corresponding marketed CFC product when given twice daily. An overall maintainment and an unchanged risk-benefit ratio has emerged for budesonide HFA given once daily, which was however slightly inferior compared with the standard twice daily regimens. PMID- 14753251 TI - Detection of Pb2+ using a hydrogel swelling microcantilever sensor. AB - Hydrogels containing benzo-18-crown-6 were used to modify microcantilevers for measurements of the concentration of Pb2+ in aqueous solutions. These microcantilevers undergo bending deflection upon exposure to solutions containing various Pb2+ concentrations as the result of a swelling of the hydrogels. It was found that a concentration of 10(-6) M Pb2+ can be detected using this technology. Other cations, such as Na+, have no effect on the deflection of this cantilever. The cation K+, which also complexes with benzo-18-crown-6, could interfere with Pb2+ detection, but only at high concentrations (> 10(-4) M). PMID- 14753250 TI - First functionalized MRI contrast agent recognizing vascular lesions. AB - A new MRI-contrast agent, EB-DTPA-Gd, that has an Evans Blue analogue as a sensing unit for endothelium lesions, was designed and synthesized. The agent also has diethylenetriamine-N,N,N',N",N"-pentaacetic acid-Gd complex (Gadolinium DTPA) units, which have been used as detection units for T1-weighted MRI. The EB DTPA-Gd was able to recognize and adsorb to the vascular endothelium-denuded region of porcine aorta, and to decrease the relaxation time of circumferential water's protons, making possible MR imaging of the endothelium-denuded region. The compound can be employed as a contrast agent for the imaging of vascular lesions using MRI. PMID- 14753252 TI - Real-time measurements of polychlorinated biphenyls using laser ionization time of-flight mass spectrometry. AB - Polychlorinated biphenyls (PCBs) are hazardous compounds for which there are few methods involving real-time measurement. Recently, the development of the pulsed laser technique has enabled us to measure aromatic hydrocarbons quite rapidly. In particular, the LI-TOFMS (laser ionization time of flight mass spectrometry) technique is expected to be a powerful method for on-line, selective and sensitive measurements. In the context of developing a highly selective and sensitive technique, the effective laser wavelength and pulse duration for laser ionization of 2-4 chlorinated PCBs are considered. Excitation at lambda = 266 nm was found to be substantially more effective than at lambda = 280, 300 or 320 nm. Also, picosecond excitation for PCBs underwent more efficient ionization (by a factor of over 10) and less fragmentation than nanosecond excitation. The achievable sensitivity for trichlorinated biphenyl in a real-time (1 min) measurement using the LI-TOFMS technique was estimated to be in the sub-ppbV range (< 0.01 mg/Nm3). PMID- 14753253 TI - Trends in atmospheric trace gas measurement instruments with membrane-based gas diffusion scrubbers. AB - A diffusion scrubber (DS) is an excellent tool for gas analysis, and there are many types of DS devices, varying both in structure and construction. In this paper, recent work on atmospheric trace gas measurements by means of DS devices are reviewed. Theoretical considerations on representative DSs are summarized first. Then, the characteristics of the key material, a gas-diffusion membrane, are discussed, and recent improvements and novel scrubbers for highly effective collection are outlined. A chromatograph is a suitable tool for the multi-gas analysis of collected species. On the other hand, solid-state fluorescence/absorbance detectors have been developed for the continuous measurement of target gases. The methods based on DS collection and subsequent detection have high sensitivities, and the detection limits can be in the low pptv levels. Accordingly, they are capable of measuring background levels, and detecting very low levels of contaminants in a cleanroom. Miniaturized advanced DS units, perhaps the next generation of DS devices, are introduced at the end of this review. DS systems have contributed significantly to our knowledge of the atmospheric dynamics and atmospheric chemistry. PMID- 14753254 TI - Distributions of major-to-ultratrace elements among the particulate and dissolved fractions in natural water as studied by ICP-AES and ICP-MS after sequential fractionation. AB - In order to elucidate the distributions of the elements among the particulate and dissolved fractions in pond water, major-to-ultratrace elements in different sizes of particles as well as in the filtrate passed through the 0.05 microm filter were determined by inductively coupled plasma atomic emission spectrometry (ICP-AES) and inductively coupled plasma mass spectrometry (ICP-MS). The different sizes of particle samples (ca. 100-300 microg each) were collected on the membrane filters with pore sizes of 10, 3.0, 1.2, 0.4, 0.2 and 0.05 microm, respectively, by sequential fractionation. As a result, about 40 elements in different sizes of particles could be determined by ICP-AES and ICP-MS, after acid digestion using HNO3/HF/HClO4. Then, the fractional distribution factors of major-to-ultratrace elements among the particulate and dissolved fractions were estimated from the analytical results. The total contents of Al, Fe, Ti, REEs (rare earth elements), Bi, Pb and Ag in the particulate fractions (larger than 0.05 microm) were more than 80-90%, while those of Ca, Sr, Cs, W, Ba, Mn and Co in the dissolved fraction, which corresponded to the filtrate passed through the 0.05 microm membrane filter, were more than 80%. It was further found that the fractional distributions of Cu and Zn in the dissolved fraction were ca. 50%. In addition, the enrichment factors (EFs) of the elements in the particulate fractions with particle sizes of 3.0-10 microm and 0.05-0.2 microm were estimated to elucidate their geochemical characteristics in natural water. PMID- 14753255 TI - Analytical approach to the discoloration of edible laver "nori" in the Ariake Sea. AB - To understand the cause of discoloration of the sea laver "nori," which is found in the Ariake Sea, the concentrations of pigments and elements in the normal and discolored laver samples were determined. In the discolored samples, a decrease in all of the pigments, chlorophyll a and carotenoids, and proteinous pigments, phycobiliproteins, was clearly observed. This was accompanied by a decrease in the content of Fe, Zn, Mn, Cu, and P. Good correlations between these elements and chlorophyll a, as well as between these elements and phycobiliproteins, were confirmed, indicating that, in addition to the deficiency of nitrogen and phosphorus, the deficiency of trace elements (Fe, Zn, Mn, and Cu), which are specifically required for photosynthesis, could be a reason for the discoloration of nori. The cause of elemental deficiency is also discussed. PMID- 14753256 TI - Contamination and biomethylation of organotin compounds in pearl/fish culture areas in Japan. AB - Uwakai of Japan is famous for pearl and yellowtail fish culture. Recently, pearl culture farming in that region has suffered from a low production of pearls. An illegal use of organotin antifouling paints on fishing nets was reported. In the line of pollution studies, thus, the present investigation was carried out to examine the contamination status and fate of organotin compounds. Totally, 23 water, 10 sediment and 8 pearl oyster tissue samples were analyzed for tributyltin (TBT), triphenyltin (TPT), and their breakdown products (di- and mono compounds) by gas chromatography combined with inductively coupled plasma mass spectrometry (GC/ICP-MS). The results show that the TBT concentrations in water, sediment and biota were in the range from 0.11 to 10.6 ng Sn l(-1), 0.35 to 2500 ng Sn g(-1), and 50.4 to 181 ng Sn g(-1), respectively. The values for sediment and biota are expressed on the dry-weight basis. Triphenyltin in water, sediment and biota were in the range from 0.009 to 0.108 ng l(-1), non-detect to 12.7 ng g(-1), and non-detect to 6.83 ng g(-1), respectively. Although the TBT concentration in seawater is below the tentative assessment level of 10 ng l(-1) set by the Japanese Environment Agency in 1992, it may cause endocrine disruption/other effects in aquatic organisms. Octyltin compounds (mono-, di- and trioctyltin) were also quantified in seawater and sediment. The detection of dibutyldimethyltin (DBDMT) and tributylmonomethyltin (TBMMT) in sediment (methylated butyltins comprised 2.8-31% of total butyltins), and TBMMT in seawater suggested that biomethylation of anthropogenic tributyltins is a significant transformation pathway in the coastal environment. PMID- 14753258 TI - Classification for dimethylarsenate-decomposing bacteria using a restrict fragment length polymorphism analysis of 16S rRNA genes. AB - A new monitoring system for bacterial communities involving dimethylarsinic acid (DMAA) decomposition was provided by combining the MPN (Most Probable Number) method and RFLP (restriction-fragment-length polymorphism analysis). The abundance of DMAA decomposing bacteria was estimated by the MPN method using a bacterial culture medium, which included DMAA as the sole carbon source, indicating bacterial cell densities of 1700 cells/ml in Lake Kahokugata and 330 cells/ml in Lake Kibagata. After isolating the dominant bacteria using agar plates, the isolates were classified into some genotype groups by RFLP analysis using 16S rDNA sequences. Classification of the RFLP analysis indicated that 14 isolates of Lake Kahokugata were classified into 6 types, which included 2 dominant types related to genus Pseudomonas, while 8 isolates of Lake Kibagata displayed 6 types including one or two isolates. Moreover, the RFLP types were unique for each lake, suggesting that DMAA decomposing bacteria were specific for the aquatic environment related to the arsenic cycle. The activities of DMAA decomposition mostly matched with the RFLP type category of the isolates. Accordingly, combining the MPN method with the RFLP analysis will play an important role in elucidating the distributions and dynamics of the DMAA decomposing bacterial community. PMID- 14753257 TI - A new luciferase reporter gene assay for the detection of androgenic and antiandrogenic effects based on a human prostate specific antigen promoter and PC3/AR human prostate cancer cells. AB - We developed a new mammalian cell-based luciferase reporter gene assay for androgenic and antiandrogenic activities of chemicals and environmental samples. Environmental samples usually have a complex matrix that may contain the constituents acting as androgen receptor (AR) agonists, AR antagonists or aryl hydrocarbon receptor (AhR) agonists. AhR agonists are known to elicit the antiandrogenic effect through cross-talk between AR and AhR signal transduction pathways. In this study, PC3/AR human prostate carcinoma cells were transiently transfected with a prostate-specific antigen (PSA) promoter-driven luciferase expression plasmid. The cells were treated with a test compound or an environmental sample for 24 h at 37 degrees C and then measured for luciferase activity. The luciferase activity was induced by dihydrotestosterone (DHT) in a concentration-dependent manner in a concentration range from 10 fM to 1 nM. R1881, a synthetic androgen receptor agonist, induced luciferase activity and its inductive effects was additive to that of DHT. The luciferase activity was not induced by cortisol, a glucocorticoid, progesterone, a progestin, and 17beta estradiol, an estrogen in a concentration range of up to 1 microM. DHT-induced luciferase activity was reduced by bicalutamide and cyproterone acetate, AR antagonists, and also by benzo[a]pyrene, an aryl hydrocarbon receptor agonist, through AhR-mediated pathways. All of these findings indicate that the present assay system correctly responds to AR agonists, AR antagonists and AhR agonist and, therefore, it is a powerful tool for the sensitive and selective screening of chemicals and environmental samples for their androgenic and antiandrogenic activities. We developed the first assay system, in which the expression of luciferase was driven by the promoter of a prostate-specific antigen gene, a typical human androgen-regulated gene. PMID- 14753259 TI - Effect of long-range transport of air mass on the ionic components in radiation fog in northern Japan. AB - Water samples from radiation and upslope types of fog were collected at Takanosu Basin and Hachimantai mountain range of Akita Prefecture in northern Japan, respectively. The effect of the long-range transport of pollutants from the Asian Continent to the basin and the mountain range on the chemical characteristics of two types of fog was studied using chemical analysis data for the water samples as well as the back-trajectories of the air mass. In particular, non-sea-salt (nss)-SO4(2-) of radiation fog provided a much higher concentration than that of upslope fog, which suggests that nss-SO4(2-) might be transported mainly from the Asian Continent. The transport and uptake mechanism of pollutants in the radiation fog water were explained based on their accumulation in the basin due to anticyclonic subsidence and by the long-lasting fog behavior. PMID- 14753260 TI - Heavy-metal pollution and its state in algae in Kakehashi river and Godani river at the foot of Ogoya mine, Ishikawa Prefecture. AB - Alga as Achnanthes minutissima among diatoms is a widely adaptable taxon on the state of an aquatic environment. In this study, it was found that diatom had a specific tolerance to heavy metals (Cu, Zn, Pb, and Cd etc.) in river water samples, because the diatom assemblage consisted of almost only Achnanthes minutissima in Kakehashi river and Godani river, which were polluted with waste water from Ogoya copper mine. The relationship between the concentrations of heavy metals (Cu, Zn, Pb and Cd) in river water and the attached substances (algae and silt etc.) and the relative abundances of diatom taxa were investigated in detail. The results indicated that the higher is the concentration of heavy metals in the river environment, the higher is only the relative abundances of Achnanthes minutissima. Thus, the taxon can be used as a bioindicator of heavy metal pollution. The relative rates of toxic chemical forms of copper in algae were 61 - 92% in the attached substances and 49-70% in the sediment on the river bed, respectively. Therefore, it was found that diatom as Achnanthes minutissima had a tolerance to heavy metals in river water, being able to live in such an environment. Since the water treated with calcium hydroxide from the deposition reservoir of Ogoya mine enters in Godani river, the river is polluted by heavy metals (Cu, Zn, Pb and Cd etc.). From the viewpoint of both biological and chemical analyses, Godani river is still polluted with heavy metals, because their concentrations in the river samples were very high. On the other hand, in Kakehashi river, the concentrations of heavy metals were very low and the distributions of some diatoms appeared in an unpolluted Nishimata river were observed. Therefore, Kakehashi river seems to be considerably recovered from heavy-metal pollution after closing the Ogoya mine. PMID- 14753261 TI - Heavy-metal distribution in river waters and sediments around a "firefly village", Shikoku, Japan: application of multivariate analysis. AB - River water and sediment samples were collected at the same site in a vicinity of an abandoned mine, and the concentrations of major elements and heavy metals were determined. The chemical correlations were observed by principal component analysis (PCA), and the samples were classified by cluster analysis (CA) based on the PCA scores. The PCA results presented a macroscopic viewpoint of covariance structure, i.e., the chemical elements could be classified into three groups: 1) major elements and heavy metals in the river water, 2) Cd, Fe and Mn in the sediments, and 3) Cu and Zn in the sediments. The CA results implied a similarity of chemical compositions in most parts of the study area, except the ranges close to the abandoned copper mine. At the mixing location of mining water with natural river water, major elements and cadmium showed simple physical mixing (conservative mixing). Other heavy metals (Cu, Fe, Mn and Zn) showed the massive precipitation at the mixing event. The PCA structure was mainly interpreted in terms of the mixing process between mining water and diluted natural river water. PMID- 14753262 TI - Concentrations of hazardous heavy metals in environmental samples collected in Xiamen, China, as determined by vapor generation non-dispersive atomic fluorescence spectrometry. AB - Non-dispersive atomic fluorescence spectrometry (NDAFS) coupled with vapor generation (VG) sample introduction was applied to the determination of the concentrations of hazardous heavy metals, such as arsenic, cadmium, lead and mercury, in seawater, soils and total airborne particulate matter (PM) collected around the Xiamen area in China. Almost 100% sample introduction efficiency was achieved by using thiourea and ascorbic acid for the pre-reduction of As(V) to As(III), K3Fe(CN)6 and tartaric acid for pre-oxidation of Pb(II) to Pb(IV), and masking the interferences arising from the co-existing transition metals to As, Cd, Hg and Pb during their vapor generation process. Moreover, a novel sample pretreatment device was developed to avoid the loss of mercury, lead, cadmium and arsenic during sample pretreatment. With such methods, the detection limit (DL) of arsenic, cadmium, lead and mercury was down to 0.08, 0.03, 0.05, 0.01 ng mL( 1) (3sigma), respectively. The relative standard deviations (RSD, n = 11) for arsenic, cadmium, lead and mercury at 10 ng mL(-1) were 0.9%, 1.6%, 1.3% and 2.0%, respectively. The concentrations of hazardous heavy metals in the environmental samples collected in Xiamen, China are in the range from 0.02 +/- 0.001 ng mL(-1) in seawater to 15.3 +/- 0.2 microg g(-1) in soils. Besides flame/GF-AAS and ICP-AES/MS, VG-NDAFS should be another choice for the determination of hazardous heavy metals in environmental samples. PMID- 14753263 TI - Detection of iron(III)-binding ligands originating from marine phytoplankton using cathodic stripping voltammetry. AB - The sample preparation and analytical methodology are described for detecting biologically produced iron(III)-binding ligands in laboratory cultures of coastal marine phytoplankton. The iron(III)-binding ligands from the culture media were purified by passage through a column packing with a hydrophobic absorbent. The concentrations and stability constants of the ligands were determined by adsorptive cathodic stripping voltammetry with competitive ligand equilibration. The analytical results of the cultivated cultures suggest that eukaryotic phytoplankton would produce iron(III)-binding ligands in analogy with other microorganisms. PMID- 14753264 TI - Determination of polymeric aluminum in soil extracted with a modified anion exchange resin as a solid-phase adsorbent by ICP-AES. AB - In the present work, a new method was established by applying solid-phase extraction (SPE) to preconcentrate and separate polymeric aluminum (Al) and using ICP-AES to determine the polymeric Al, the total monomeric Al, and the total Al in soil extracts, respectively. A modified resin was prepared with impregnated 8 hydroxyquinoline-5-sulfoxinate (HQS) on the anion-exchange resin. It has good recognition ability for Al fractions, compared to the commonly used cation ion exchange resin, which has a better ability to adsorb cations and a weak ability to recognize detailed Al species. The optimum conditions for Al fractionation sorption, elution and separation and the interference of foreign ions were studied with the prepared resin by continuous column and batch procedures. Monomeric Al was bound to Pyrocathecol Violet (PCV) at pH 6.2, whereas the polymeric Al species did not react with PCV for at least 15 min. Because a stable complex of Al-PCV was not absorbed on the HQS modified resin, the polymeric Al could be preconcentrated on-line by the HQS-modified resin. The adsorbed polymeric Al was eluted with 3 mL of 3 mol L(-1) of HCl, and then detected by ICP AES. The method has been applied to directly determine polymeric Al in soil extracts with high selectivity as well as a high preconcentration factor. It gives a limit of detection of 0.6 ng mL(-1) with a relative standard deviation of less than 5.7% (n = 5, 0.24 microg mL(-1) Al). PMID- 14753265 TI - Derivatization of tributyltin with sodium tetrakis(4-fluorophenyl)-borate for sensitivity improvement of tandem mass spectrometry. AB - Derivatization of tributyltin for tandem mass spectrometry is described. Tributyltin (TBT) and triphenyltin (TPT) were derivatized with sodium tetrakis(4 fluorophenyl)borate. After optimization of their MS/MS conditions, derivatization conditions were examined. Under the optimum conditions using in-situ derivatization, the calibration curves for the TBT and TPT were linear in the ranges of 0.4 - 200 and 1.2 - 200 pg of Sn, respectively. The detection limits for TBT and TPT were 0.07 and 0.43 pg of Sn, respectively. In the case of TBT, the detection limit with 4-fluorophenylation was improved about five times compared with that with pentylation (0.35 pg). This improvement is ascribed to the bond-dissociation energy of Sn-aryl being stronger than that of Sn-alkyl. Namely, the selective fragmentation of 4-fluorophenyl TBT resulted in high sensitivity. The relative recoveries of TBT and TPT from seawater were 99 and 109%, respectively. The method was successfully applied to the seawater samples. PMID- 14753266 TI - Simple analysis of volatile organic compounds (VOCs) in the atmosphere using passive samplers. AB - A simple analysis of volatile organic compounds (VOCs), such as benzene, toluene, m,p-xylene, and o-xylene, at low levels in the atmosphere was conducted using passive samplers. The methods were applied to analyzing the behavior and origin of VOCs in Kyoto City. The passive samplers were exposed for 7 - 14 days at sampling sites in Kyoto City and for 30 days in the mountains (Mt. Hiei and Mt. Daimonji). Shibata gas-tube samplers packed with activated carbon were used for the determination of VOCs. The absorbed VOCs were extracted into carbon disulfide (CS2) and measured by FID-GC. The determination limits and relative standard deviations for VOCs were 0.3 microg/m3 and 3%, respectively. The samplers were set up at 5 sites in March, 2001 and at 13 stations on Mt. Hiei in November, 2002. The average concentrations of ambient benzene, which were higher than the environmental criterion (3.0 microg/m3), except for those on Mt. Daimonji from March, 2001, to February, 2002, decreased to below 3.0 microg/m3 from March, 2002, to February, 2003. The decrease in ambient benzene may have been due to a decrease in the benzene content in gasoline by the end of 1999, and also by implementation of the Pollutant Release and Transfer Register (PRTR) Act in 2001. PMID- 14753267 TI - On-line collection/concentration and detection of sulfur dioxide in air by flow injection spectrophotometry coupled with a chromatomembrane cell. AB - A simple and rapid procedure for SO2 determination in air was developed by using a flow injection analysis (FIA) system coupled with a 3-hole chromatomembrane cell (CMC). The CMC was applied for the on-line collection/concentration of SO2 from air into a solution of 2 g l(-1) triethanolamine (TEA) solution as an absorbing solution: SO2 was converted to SO3(2-) in the alkaline absorbing solution. The solution containing absorbed SO2 was introduced into the carrier stream of the FIA system. The amount of SO3(2-) in the absorbing solution was measured by spectrophotometry with a mixed reagent of pararosaniline and formaldehyde, and was converted to the concentration of SO2 in the air sample. A calibration graph prepared by using standard sodium sulfite aqueous solutions was adopted for the determination of SO3(2-) in the absorbing solution. The SO2 concentration in indoor air examined was found to be 22.7 +/- 0.2 ppbv using 20 ml of air sample with the air flow rate of 5 ml min(-1), where the relative standard deviation was 1.7%. The detection limit for aqueous solutions and air samples were 6.9 x 10(-8) M and 0.48 ppbv, respectively. The measuring time for one sample was about 10 min when a 20 ml air sample was used. The interferences from common anionic species, formaldehyde and acetaldehyde, were also examined. PMID- 14753268 TI - Determination of atmospheric nitrobenzanthrones by high-performance liquid chromatography with chemiluminescence detection. AB - A method using high-performance liquid chromatography with chemiluminescence detection was developed for analyzing mutagenic nitrobenzanthrone (NBA) isomers in airborne particulates. The method was a modification of our previously described method for analyzing nitropolycyclic aromatic hydrocarbons (NPAHs). The pretreatment and reducing conditions for 1-, 2-, 3- and 10-NBAs were the same as those for NPAHs. In order to separate these NBA isomers, we used a polymeric-type ODS column (Cosmosil 5C-18MS); a mixture of 40% acetonitrile and 60% 10 mM imidazole-HClO4 buffer was employed as the mobile phase at a flow rate of 1 mL/min. The isomers of 1-, 2-, 3- and 10-NBA were determined in chemiluminescence with linear calibration graphs from 0.1 to 4 pmol, from 200 to 4000 pmol, from 1 to 50 pmol and from 10 to 400 pmol, respectively. The detection limits (S/N = 3) of 1-, 2-, 3- and 10-NBA isomers were 0.02 pmol, 35 pmol, 0.3 pmol and 3 pmol, respectively. The method was used to analyze airborne particulates at a heavy traffic site in Kanazawa. 2- and 3-NBAs were detected in the extracts of the particulates, while 1-NBA and 10-NBA were not detected. The atmospheric concentrations of 2- and 3-NBAs were 1.83 pmol/m3 and 24.7 fmol/m3, respectively. PMID- 14753269 TI - Ion chromatographic determination of ammonia in air using a sampling tube of porous carbon. AB - The ion chromatographic determination of ammonia in air using a sampling tube of porous carbon carbonized at 500 degrees C was examined. When the mean recovery and the reproducibility for a series of five determinations were examined for 1 and 10 ppm ammonia gases, the mean recovery (n = 5) and the relative standard deviation were 97.0% and 3.5% for 1 ppm and 86.9% and 2.8% for 10 ppm, respectively. Furthermore, the recovery from 10 ppm ammonia gas increased with an increase in the extraction time, and a recovery of 99.7% was obtained for 180 min of extraction time. The mean collection efficiency for 1 and 10 ppm ammonia gas was 102.5% and 96.5%, respectively. The relation between the sampling volume and the peak area was linear, and the linearity was 0.999 of the correlation coefficient. The ammonia gas concentration in an actual goat shed could be determined by this sampling device for a sampling volume of 5 L at a flow rate of 1 L/min; 0.79 ppm of the determination value practically agreed as compare with 0.78 ppm from collection by a boric acid solution. PMID- 14753270 TI - Determination of hydroxylated polycyclic aromatic hydrocarbons in airborne particulates by high-performance liquid chromatography with fluorescence detection. AB - This paper describes a highly sensitive and selective method for the determination of hydroxylated polycyclic aromatic hydrocarbons (PAHs-OH) in airborne particulates. PAHs-OH were extracted from airborne particulates with ethanol by ultrasonication. The extractant was further cleaned up by solid phase extraction (SPE) with an aminopropylsilica cartridge, and subsequently analyzed by reversed-phase high-performance liquid chromatography with fluorescence detection. 2-Hydroxy-1-acetonaphthone was used as an internal standard. By the proposed method, 2-hydroxyfluorene and 1-hydroxypyrene were identified in airborne particulates and their concentrations were determined for the first time. PMID- 14753271 TI - Improved detectability with a polymer-based trapping device in rapid HPLC analysis for ultra-low levels of bisphenol A (BPA) in environmental samples. AB - A new high-performance liquid chromatography (HPLC) method has been developed to detect ultra-low concentrations of bisphenol-A (BPA) (below 1 ng/L (ppt)) using column switching electrochemical detection (ECD). The results were superior to those obtained from manual pretreatment procedure with membrane stationary phase. BPA is inherently ubiquitous in the environment, including tools and solvents used for its analysis; to obtain meaningful results, therefore, the concentration of the overall BPA contamination must be below the detection limit for BPA using the analytical system. Therefore, purified water for preparing the standard BPA solution must be filtered with a hydrophobic membrane to suppress BPA background levels of contamination. In addition, we investigated methods for effectively preserving environmental water containing BPA. The addition of a small amount of ethylenediaminetetraacetic acid (EDTA) provided good recovery even after overnight storage. By employing these precautionary measures and procedures to reduce BPA contamination from the analytical procedure, we could accurately determine l(-10) ppt of BPA in environmental water samples using a column switching HPLC system. PMID- 14753272 TI - A simple and sensitive method for the determination of hydroxylamine in fresh water samples using hypochlorite followed by gas chromatography. AB - A new and simple method for the determination of hydroxylamine in environmental water, such as fresh rivers and lakes using hypochlorite, followed by its gas choromatographic detection, has been developed. A glass vial filled with sample water was sealed by a butyl-rubber stopper and aluminum cap without head-space, and then sodium hypochlorite solution was injected into the vial through a syringe to convert hydroxylamine to nitrous oxide. The head-space in the glass vial was prepared with 99.9% grade N2 using a gas-tight syringe. After the glass vial was shaken for a few minutes, nitrous oxide in the gas-phase was measured by a gas chromatograph with an electron-capture detector. The dissolved nitrous oxide in the liquid-phase was calculated according to the solubility formula. The proposed method was applied to the analysis of fresh-water samples taken from Iu river and Hii river, flowing into brackish Lakes Nakaumi and Shinji, respectively. PMID- 14753273 TI - Characterization and determination of linear alkylbenzenesulfonates in environmental water samples by high-performance liquid chromatography with a hydrophilic polymer column and electrospray ionization mass spectrometric detection. AB - A liquid chromatography-mass spectrometry (LC/MS) method was developed for the separation and determination of linear alkylbenzenesulfonates (C10-C14 LAS) in environmental water samples using a hydrophilic polymer column (Shodex Mspak GF 310 4D). This method involves a solid-phase extraction of the LAS samples with a Sep-Pak PS-2 cartridge. The LAS components were separated on the column with a mobile phase of 29% (w/v) acetonitrile-water containing 0.8 mM di-n-butylammonium acetate and 0.2 M acetic acid, and were detected by mass spectrometry with electrospray ionization. Detection limits of the developed method based on selected ion monitoring (SIM) technique for the C10-C14 LAS standards were 13-47 ng L(-1). The concentrations of the C10-C14 LAS in the environmental water samples ranged between 5-317 microg L(-1) for a river water sample and 0.4-6.4 microg L(-1) for a seawater sample. Linear relationships between the logarithms of retention factors and the alkyl chain lengths for each phenyl positional isomer of LAS could successfully be used for the identification of the isomer peaks. PMID- 14753274 TI - Micro-organic ion-associate phase extraction via in situ fresh phase formation for the preconcentration and determination of di(2-ethylhexyl)phthalate in river water by HPLC. AB - A high-enrichment method was proposed for the HPLC determination of trace di(2 ethylhexyl)phthalate (DEHP) in environmental water. A micro-organic ion-associate phase (IAP) was formed in situ from an aqueous sample by adding 4 trifluoromethylanilinium ion and dodecylbenzenesulfonate ions. Centrifugation of the solution led to the isolation of a liquid organic phase containing DEHP at the bottom of the centrifuge tube. The volume of the phase formed was less than 30 microL. DEHP was extracted into the IAP quantitatively during phase formation. After discarding the aqueous phase, the ion associate was dissolved with 50 microL of 2-methoxyethanol, and DEHP in the concentrate was determined by HPLC with an ultra-violet (UV) diode-array detector. DEHP in the concentrations range from 0.8 to 78 microg L(-1) was determined with good precision. The recovery tests for DEHP added to some river water were satisfactory. The detection limit of DEHP, defined as 3-times the standard deviation of the blank signals, was 0.07 microg L(-1) (n = 3). The present method is very simple, and was applied to the determination of DEHP in the river water samples collected around Toyama City, Japan. PMID- 14753275 TI - Determination of hydroxyl radical photoproduction rates in natural waters. AB - The photochemical formation rates of hydroxyl radicals (OH radicals) in river water and seawater were determined by a simple, rapid and sensitive benzene probe method, in which phenol formed by the reaction between benzene and photochemically-generated OH radicals was analyzed by on-line preconcentration HPLC. The OH radical formation rates from well-known OH radical sources, such as nitrate, nitrite and hydrogen peroxide, were in good agreement with those reported previously. River water samples containing high concentrations of nitrate and nitrite were found to show high OH radical formation rates. Ten to 80% of the OH radical formation in river water and seawater was due to the photolysis of nitrate and nitrite, but OH radical formation from hydrogen peroxide was negligible. The OH radical formation from unknown sources other than nitrate, nitrite and hydrogen peroxide was strongly correlated to the amount of fluorescent matter. PMID- 14753276 TI - Determinations of humic substances and other dissolved organic matter and their effects on the increase of COD in Lake Biwa. AB - Humic substances and other dissolved organic matter (DOM) in Lake Biwa and the surrounding rivers were investigated to elucidate their origins and behavior. An annual increase in chemical oxygen demand (COD) has been observed in the northern basin of Lake Biwa since 1985. The concentrations of dissolved organic carbon (DOC) in the northern and southern basins of Lake Biwa were 1.7-2.4 mgC/l and 1.9 2.6 mgC/l, respectively. The DOC concentrations tended to be high in summer and low in winter, and the seasonal changes in the concentrations of humic substances were small. The humic substances content of DOM was considered to be comparatively small because the ratio of the concentration of humic substances to DOC was in the range of 0.14-0.32. From the results of the fractionation of DOM in lake waters, it was estimated that hydrophobic acids, such as humic substances and hydrophilic acids, were about 25% and 45%, respectively. The main origin of hydrophobic acids in Lake Biwa may be humic substances from soils around the rivers that flow into Lake Biwa, while hydrophilic acids may be due to the inner production by phytoplankton. Therefore, the increase of COD in the northern basin of Lake Biwa may be attributed to the contributions of not only humic substances but also hydrophilic acids. PMID- 14753277 TI - A simple in situ visual and tristimulus colorimetric method for the determination of trace arsenic in environmental water after its collection on a mercury(II) impregnated paper. AB - A simple in situ visual and tristimulus colorimetric method for the determination of trace arsenic in environmental water after collecting arsenic on a test paper impregnated with mercury(II) bromide and rosaniline chloride by its reduction aeration has been developed. The color development on the test paper is based on the formations of AsH(HgBr)2 (yellow) and/or As(HgBr)3 (brownish yellow) by a reaction between mercury(II) bromide and arsine (AsH3), which is produced through the reduction of As(III) (arsenite ion) and/or As(V) (arsenate ion) in a sample solution. To a sample solution, potassium iodide, tin(II) chloride, zinc sand and 4 ml of 6 M hydrochloric acid solution were added successively. The liberated arsine was collected on the test paper. The yellow or brownish-yellow color intensity on the test paper was measured by a tristimulus colorimeter and also by a visual method. The established method is applicable to the determination of arsenic in environmental water sample such as river, brackish, and seawater types. PMID- 14753278 TI - Successive determination of copper and iron by a flow injection-catalytic photometric method using a serial flow cell. AB - A flow injection-catalytic spectrophotometric method using a serial flow cell was proposed for the successive determination of trace amounts of copper and iron. This method is based on the oxidation coupling of p-anisidine with N,N dimethylaniline in the presence of hydrogen peroxide to form a dye, which has an absorption maximum at 740 nm. In this indicator reaction, ligands such as 1,10 phenanthroline (phen) and diphosphate were achieved to improve the sensitivity and selectivity. Under the optimal experimental conditions, the determinable ranges were 0.05-5 ppb for copper and 0.5 - 100 ppb for iron, respectively. The RSDs (n = 10) were 0.78% for 0.5 ppb copper(II) and 0.5% for 200 ppb iron(III). The sample throughput was 30 h(-1). The present flow-injection method was applied to the determination of copper and iron in standard river water, tap water, and other natural water samples, and also to the analysis of labile and inert complexes in synthesized samples containing humic acid with copper(II) or iron(III). PMID- 14753279 TI - Preconcentration of trace aluminum from environmental water samples with 5' chloro-5-dodecyl-2,4,2'-trihydroxyazobenzene impregnated macroporous resin. AB - 5'-Chloro-5-dodecyl-2,4,2'-trihydroxyazobenzene (CDTA) impregnated XAD-4 resin (CDTA resin) was prepared, and applied to the preconcentration of trace aluminum from environmental water samples with satisfactory results. Aluminum was measured by flow-injection analysis (FIA) with fluorescence detection by using lumogallion after the preconcentration of aluminum with CDTA resin. Fe3+ and other coexisting substances had no interference on the determination of aluminum by the present method. Chemical speciation of trace aluminum in environmental water was carried out by using the CDTA resin after the separation of aluminum species with ionic exchangers without changing the pH. About 50% of water-soluble aluminum in Lake Biwa was found to be aluminum organic species (anion species). PMID- 14753280 TI - Determination of heavy metals and rare earth elements in environmental samples by ICP-MS after solid phase preconcentration with chelating resin fibers and anion exchanger filters. AB - A solid phase collection/concentration method using anion exchanger filters and a small syringe packed with chelating resin fibers is adopted as a preconcentration tool for trace elements and a separation tool for matrices in aqueous samples prior to the measurement by inductively coupled plasma-mass spectrometry (ICP MS). The effects of fiber volume, sample volume, eluent volume, and sample flow rate on metal recoveries were investigated in detail to obtain optimum pretreatment conditions. Several heavy metals (HMs) such as, V, Mn, Co, Ni, Cu, Zn, Ga, Cd, Pb, Th and U, as well as 14 rare earth elements (REEs) in sample solutions at pH 6 were quantitatively collected on the solid phase. These adsorbed elements were completely recovered by eluting with 2 ml of 1.0 M nitric acid. At pH 6, more than 99% of alkali and alkaline earth metals in sample solutions were eliminated. The proposed method was evaluated by analyzing two standard reference materials (SRM): peach leaves (NIST 1547) and pond sediment (NIES No. 2). The solid samples were decomposed by microwave-heating and pressurizing acid digestion technique, and then treated by the proposed syringe type pretreatment method, followed by the ICP-MS measurement. The analytical results for HMs in the SRMs obtained by the present method agreed well with the certified values. PMID- 14753281 TI - Partitionings and kinetic behaviors of major-to-ultratrace elements between industrial waste incineration fly and bottom ashes as studied by ICP-AES and ICP MS. AB - The partitionings of major-to-ultratrace elements between industrial waste incineration fly ash (IWIFA) and industrial waste incineration bottom ash (IWIBA) in industrial waste incinerators were investigated by measuring their concentration distributions, where the incineration ash samples were collected from three different types of industrial waste incinerators. The concentrations of the elements in the incineration ash samples were determined by inductively coupled plasma atomic emission spectrometry (ICP-AES) and inductively coupled plasma mass spectrometry (ICP-MS). As a result, ca. 40 elements in the concentration range from mg g(-1) to sub-microg g(-1) could be determined in both IWIFA and IWIBA samples. The concentration ratios of CF/CB (CF, concentration in fly ash; CB, concentration in bottom ash) for analyte elements were used to evaluate the partitionings of the elements between fly and bottom ashes. Then, the correlations between the CF/CB values of the elements and the dissociation energies of their monoxides were examined to evaluate the kinetic behaviors of the elements during the incineration processes. It was found that lithophile and siderophile elements, which have a large affinity with oxygen, were almost equally distributed between fly and bottom ashes, regardless of the dissociation energies of their monoxides. On the other hand, chalcophile elements with rather large volatility provided different behaviors; the elements with the smaller dissociation energies of monoxides were more partitioned in fly ashes than those with the larger ones. PMID- 14753282 TI - Lead isotopic compositions of environmental certified reference materials for an inter-laboratory comparison of lead isotope analysis. AB - Lead isotope ratios, viz. 207Pb/206Pb and 208Pb/206Pb, of the commercially available certified reference materials (CRMs) issued in Japan are presented with an objective to provide a data set, which will be useful for the quality assurance of analytical procedures, instrumental performance and method validation of the laboratories involved in environmental lead isotope ratio analysis. The analytical method used in the present study was inductively coupled plasma quadrupole mass spectrometry (ICPQMS) preceded by acid digestion and with/without chemical separation of lead from the matrix. The precision of the measurements in terms of the relative standard deviation (RSD) of triplicated analyses was 0.19% and 0.14%, for 207Pb/20Pb and 208Pb/206Pb, respectively. The trueness of lead isotope ratio measurements of the present study was tested with a few CRMs, which have been analyzed by other analytical methods and reported in various literature. The lead isotopic ratios of 18 environmental matrix CRMs (including 6 CRMs analyzed for our method validation) are presented and the distribution of their ratios is briefly discussed. PMID- 14753283 TI - Determination of ortho-phenylphenol residues in lemon rind by high-performance liquid chromatography with electrochemical detection using a microbore column. AB - A simple and highly sensitive method has been developed for determining ortho phenylphenol (OPP) in lemon rind by high-performance liquid chromatography with electrochemical detection using a microbore column (microHPLC-ECD). Based on the voltammetric behavior of OPP, microHPLC-ECD was established using a CAPCELL PAK C 18 UG 120 microbore ODS column, 17 mM acetic acid-sodium acetate buffer (pH 4.0)/acetonitrile (60/40, v/v) as a mobile phase and an applied potential at +0.9 V vs. Ag/AgCl. The current peak height was found to be linearly related to the amount of OPP injected from 3.4 pg to 1.7 ng (r > 0.999). The detection limit (S/N = 3) was 3.4 pg (20 fmol), which was 100 times greater in terms of sensitivity when compared to conventional HPLC with UV detection. Standard OPP at 0.425 ng was detected with a relative standard deviation (RSD) of 1.9% (n = 10). The OPP contents in several lemon samples were determined by the present method. The recoveries of OPP from lemon rind exceeded 98% with an RSD (n = 5) of less than 3.01%. PMID- 14753284 TI - A miniaturized FIA system for the determination of residual chlorine in environmental water samples. AB - The present study was aimed at developing a portable miniaturized FIA system with a two-electrode based amperometric detector for on-site monitoring of residual chlorine in environmental samples. The electrode potential control and current data collection were conducted with a home-designed potentiostat with a size of 150 x 100 x 40 mm and the weight of 350 g. It was confirmed that the amperometric response increased in proportion to the chlorine concentration up to 5 microg/ml, and the detection limit for chlorine was 0.05 microg/ml. Besides a drastic reduction in the size of the analytical system, such miniaturization achieved the advantages of increased speed, minimal mobile-phase consumption, and high sensitivity. The proposed method was successfully applied to the determination of residual chlorine in samples of pool water and tap water. The results were well agreed with those obtained by the N,N-diethyl-p-phenylenediamine (DPD) photometric method. PMID- 14753285 TI - Fluorometric determination of sulfite and nitrite in aqueous samples using a novel detection unit of a microfluidic device. AB - On-chip fluorescence determination of sulfite and nitrite with N-(9 acridinyl)maleimide (NAM) and 2,3-diaminonaphthalene (DAN) has been developed using a novel fluorescence detection unit for microchip analysis. Usually, these fluorescence reagents are derivatized and detected separately in microchip analysis because different fluorescence wavelengths are emitted. The proposed fluorescence detection unit has optical fibers with no optical filter, and plural wavelengths of fluorescence were detected sensitively, even in the microchip. In this study, the simultaneous determination of sulfite and nitrite in environmental samples was performed with a polymer microchip analysis system. The calibration curves of sulfite and nitrite showed linear relations (R2 = 0.998 (sulfite) and R2 = 0.990 (nitrite)), and the relative standard deviations (RSD) for 4 runs were 2.1% (20 microM sulfite) and 1.3% (20 microM nitrite), respectively. The proposed method was applied to the recovery test of sulfite and nitrite in environmental samples. PMID- 14753286 TI - Atmospheric concentrations of sulfur dioxide, nitrogen oxides, ammonia, hydrogen chloride, nitric acid, formic and acetic acids in the south of Vietnam measured by the passive sampling method. AB - A passive sampling method was applied to measure the concentrations of air pollutants, such as sulfur dioxide, nitrogen oxides, ammonia, hydrogen chloride, and nitric, formic and acetic acids, in the ambient air at four test sites in the southern region of Vietnam. The monthly averages and the average concentrations of air pollutants during the period from July 2001 to September 2002 are reported here. The concentrations of air pollutants varied widely, depending on the test sites and the sampling periods. The average concentrations of sulfur dioxide in the air during the period from July 2001 to September 2002 at the four test sites were 3.3-16.9 ppb, and those of nitrogen dioxide were 5.4-12.8 ppb. The concentrations of nitrogen monoxide and hydrogen chloride were very low at all of the test sites. The observed concentrations of all of the above-mentioned pollutants were lower than those of the Vietnamese standards of air pollutants. PMID- 14753287 TI - Determination of carbon dioxide and acid components in exhaust gas by suppressed ion chromatography. AB - Although anions are usually determined by suppressed ion chromatography (IC), carbonate and bicarbonate ions can not be determined, because a mixed solution of sodium carbonate and sodium hydrogencarbonate is used as the eluent. This paper describes an IC method for the determination of carbonate ion and common anions using an IonPac AG17/AS17 column, an EG 40 eluent generator and a conductivity detector. The proposed IC method could determine carbonate ion and anions within 6 min. The relative standard deviations (n = 5) for chloride (0.4 mg L(-1)), nitrite (0.8 mg L(-1)), carbonate (100 mg L(-1)), nitrate (1.0 mg L(-1)) and sulfate (2.0 mg L(-1)) ions were 5.1%, 1.1%, 4.2%, 5.1% and 1.1%, respectively. In addition, the absorbing solution of carbon dioxide was examined, and 2-amino-2 methyl-1-propanol was found to be a good absorbing solution. The proposed IC method was applied to the determination of carbon dioxide and acid components in flue gas and automobile exhaust gas. PMID- 14753288 TI - An improved method for the analysis of dimethyl sulfoxide in water samples. AB - An analytical method for dimethyl sulfoxide (DMSO) in aqueous samples at nanomolar levels has been improved. DMSO was reduced to dimethyl sulfide (DMS), concentrated on an adsorbent, and measured by gas chromatography. In the presence of iron chloride, the sodium borohydride (NaBH4) reduction of DMSO proceeded smoothly and efficiently, and the repeatability of this reaction was significantly improved. The detection limit was 0.27 nM for DMSO, and its repeatability of the peak-area measurement was 4.1% as RSD (n = 5). PMID- 14753289 TI - Determination of triazine pesticides and related compounds in environmental water by liquid chromatography-mass spectrometry. AB - A method for the determination of 5 triazine herbicides and 12 degradation products in environmental water samples using liquid chromatography-electrospray ionization mass spectrometry (LC/ESI/MS) has been developed. The pesticides in water were extracted with two types of solid phase: a styrene-divinylbenzene copolymer and a graphitized carbon black. Desorption solvents for the extracted compounds were acetone for the styrene-divinylbenzene copolymer and methanol for the graphitized carbon black. Overall recoveries from ground water and river water ranged from 73% to 111%. The limits of detection (LODs) were 0.2 to 28 ng l(-1). This method was applied to several ground water samples. PMID- 14753290 TI - Ultrafiltration and alumina adsorption of micelles for the preconcentration of copper(II) in water. AB - Micellar-enhanced ultrafiltration was considered from the viewpoint of trace analysis, by taking copper(II) as an example. Copper(II) cations were collected electrostatically on micelles of sodium dodecyl sulfate, and separated from the mother liquor by ultrafiltration. However, the final solution contained a large amount of surfactant, which caused serious interference in the determination. This problem was overcome by using alumina adsorption, where negatively charged micelles were adsorbed on positively charged alumina particles and then recovered by conventional filtration. The copper was leached from the micelles with 4 mol l(-1) nitric acid, leaving the surfactant on the alumina. The proposed method was successfully applied to an analysis of certified reference water samples. PMID- 14753291 TI - Ion-pair formation of a copper(II)-ammine complex with an anionic surfactant and the recovery of copper(II) from ammonia medium by the surfactant-gel extraction method. AB - The extraction and separation of copper(II), zinc(II), cobalt(II), and cadmium(II) were investigated. Both copper(II) and zinc(II) formed ammine complexes, while cadmium(II) and cobalt(II) formed hydroxide precipitates in an ammonia medium. By the addition of sodium dodecylsulfate (SDS), a copper(II) complex formed an ion-pair (copper-ammine-DS), which was extracted into the SDS phase. However, a zinc(II) complex did not form an ion-pair, and was soluble in water. Copper(II) ion was recovered by stripping (back-extraction) after the addition of hydrochloric acid. This method was applied to the separation of copper(II) in a brass alloy. PMID- 14753292 TI - Preparation of artificially spiked soil with polycyclic aromatic hydrocarbons for soil pollution analysis. AB - To confirm the method for preparing artificially spiked soil with polycyclic aromatic hydrocarbons (PAHs), we tested the homogeneity of PAHs in spiked soils, which were prepared by three different procedures, by using kaolin and ando soil. When the slurry of kaolin and acetone containing PAHs were evaporated by a rotary evaporator at 30 - 35 degrees C, the most homogeneous distribution of PAHs was obtained in the spiked soil. This procedure was applied to the preparation of PAH spiked soil for natural soil (ando soil). Such spiked soils can be useful as the standard materials for standardization of the analytical methods for PAHs in the soil and sediment samples. PMID- 14753293 TI - Quantitative mRNA analysis of eight bovine 5-HT receptor subtypes in brain, abomasum, and intestine by real-time RT-PCR. AB - Serotoninergic pathways are involved in economically important bovine gastrointestinal (GI) motility disorders such as displaced abomasum and cecal dilatation/dislocation. The existing research tools to investigate the role of serotoninergic pathways in such disorders in ruminants comprise functional pharmacological methods, e.g., in vitro contractility studies in tissue baths, and electromyographical recordings in vivo. However, no tools for quantification of bovine serotonin receptor [5-hydroxytryptamine receptor (5-HTR)] expression were available so far. This study aimed to develop real-time RT-PCR assays for quantitative mRNA analysis of bovine 5-HTR subtypes. Because the bovine 5-HTR coding sequences (CDSs) were completely unknown, multiple species (human, mouse, and rat) alignment of complete CDS was used for primer design in highly homologous regions. LightCycler real-time RT-PCR assays (partial CDS) for the following bovine 5-HTR subtypes were developed and validated: 5-HTR1A, 5-HTR1B, 5 HTR1D, 5-HTR1F, 5-HTR2A, 5-HTR2B, 5-HTR2C, and 5-HTR4. Intra- and inter-assay coefficients of variation (CV) for the eight established assays were small, ranging from 0.49% to 2.46%. As a first physiological application, 5-HTR mRNA expression levels were measured in brain, abomasum, and intestine of 10 healthy, lactating dairy cows. The 5-HTR expression was quantified by normalization to the housekeeping gene glyceraldehyde-phosphate-dehydrogenase (GAPDH). The 5-HTR subtype expression levels ranged from 0.001% (5-HTR2C in intestine) to 1% 5 HTR/GAPDH (5-HTR1B and 5-HTR4 in intestine). There were high variations of 5-HTR subtype mRNA expression within tissues across receptor subtypes and within receptor subtypes across tissues. In conclusion, accurate real-time RT-PCR assays for quantitative analysis of bovine 5-HTR subtype gene expression were developed and validated. PMID- 14753294 TI - The expression of urotensin II receptor (U2R) is up-regulated by interferon gamma. AB - Urotensin-II (U-II) was identified as the natural ligand of the G protein-coupled receptor GPR14, which has been correspondingly renamed Urotensin-II receptor (U2R). The tissue distribution of U2R and the pharmacological effects of U-II suggest a novel neurohormonal system with potent cardiovascular effects. We here report the human rhabdomyosarcoma cell line TE-671 as the first natural and endogenous source of functional U2R in an immortalized cell line. In TE-671 cells, U-II stimulated extracellular signal regulated kinase phosphorylation and increased c-fos mRNA expression. Furthermore, we demonstrate that the expression of U2R mRNA and functional U-II high affinity binding sites are serum-responsive and that they are specifically up-regulated by interferon gamma (IFNgamma). We propose that IFNgamma contributes to the previously observed increase of U2R density in the heart tissue of congestive heart failure (CHF) patients and we suggest that U2R up-regulation, as a consequence of an inflammatory response, could lead to a clinical worsening of this disease. PMID- 14753295 TI - Phage display of random peptide libraries: applications, limits, and potential. AB - The identification of ligands from large biological libraries by phage display has now been used for almost 15 years. Most of the successful reports on high affinity ligand identification originated from work with different antibody libraries. In contrast, the progress of applying phage display to random peptide libraries was relatively slow. However, in the last few years several improvements have led to an increasing number of published peptide ligands identified by phage display from such libraries and which exhibited good biological activity and high affinity. This review summarizes the current state and the technical progress of the application of random peptide libraries using filamentous phage for ligand identification. PMID- 14753296 TI - Novel cell line selectively expressing neuropeptide Y-Y2 receptors. AB - Neuropeptide Y (NPY) recognition by the human neuroblastoma cell lines SiMa, Kelly, SH-SY5Y, CHP-234, and MHH-NB-11 was analyzed in radioactive binding assays using tritiated NPY. For the cell lines CHP-234 and MHH-NB-11 binding of [3H]propionyl-NPY was observed with Kd-values of 0.64 +/- 0.07 nM and 0.53 +/- 0.12 nM, respectively, determined by saturation analysis with non-linear regression. The receptor subtype was determined by competition analysis using the subtype selective NPY analogues [Leu31, Pro34]-NPY (NPY-Y1, NPY-Y5), [Ahx(5-24)] NPY (NPY-Y2), [Ala31, Aib32]-NPY (NPY-Y5), NPY [3-36] (NPY-Y2, NPY-Y5), and NPY [13-36] (NPY-Y2). Both cell lines, CHP-234 and MHH-NB-11, the latter one being characterized for NPY receptors for the first time, showed exclusive expression of NPY-Y2 receptors. In both cell lines binding of NPY induced signal transduction, which was monitored as reduction of forskolin-induced cAMP production in an ELISA. PMID- 14753297 TI - Tautomerism in computer-aided drug design. AB - Tautomers are often disregarded in computer-aided molecular modeling applications. Little is known about the different tautomeric states of a molecule and they are rarely registered in chemical databases. Tautomeric forms of a molecule differ in shape, functional groups, surface, and hydrogen-bonding pattern. Calculation of physical-chemical properties and molecular descriptors differ from one tautomeric state to the other as it is demonstrated with an example of the log P calculation, similarity index, and the complementarity pattern to the targeted protein. Considering tautomery in ligand-protein interactions therefore has a significant impact on the prediction of the ligand binding using various docking techniques. This article points on hitherto unaddressed issue of tautomerism in computer-aided drug design. PMID- 14753298 TI - Is your interventional radiology service ready for SARS?: The Singapore experience. AB - The recent epidemic of severe acute respiratory syndrome caught many by surprise. Hitherto, infection control has not been in the forefront of radiological practice. Many interventional radiology (IR) services are therefore not equipped to deal with such a disease. In this review, we share our experience from the interventional radiologist's perspective, report on the acute measures instituted within our departments and explore the long-term effects of such a disease on the practice of IR. PMID- 14753299 TI - Informed consent for interventional radiology procedures: a survey detailing current European practice. AB - PURPOSE: Official recommendations for obtaining informed consent for interventional radiology procedures are that the patient gives their consent to the operator more than 24 hr prior to the procedure. This has significant implications for interventional radiology practice. The purpose of this study was to identify the proportion of European interventional radiologists who conform to these guidelines. METHODS: A questionnaire was designed consisting of 12 questions on current working practice and opinions regarding informed consent. These questions related to where, when and by whom consent was obtained from the patient. Questions also related to the use of formal consent forms and written patient information leaflets. Respondents were asked whether they felt patients received adequate explanation regarding indications for intervention, the procedure, alternative treatment options and complications. The questionnaire was distributed to 786 European interventional radiologists who were members of interventional societies. The anonymous replies were then entered into a database and analyzed. RESULTS: Two hundred and fifty-four (32.3%) questionnaires were returned. Institutions were classified as academic (56.7%), non-academic (40.5%) or private (2.8%). Depending on the procedure, in a significant proportion of patients consent was obtained in the outpatient department (22%), on the ward (65%) and in the radiology day case ward (25%), but in over half (56%) of patients consent or re-consent was obtained in the interventional suite. Fifty percent of respondents indicated that they obtain consent more than 24 hr before some procedures, in 42.9% consent is obtained on the morning of the procedure and 48.8% indicated that in some patients consent is obtained immediately before the procedure. We found that junior medical staff obtained consent in 58% of cases. Eighty-two percent of respondents do not use specific consent forms and 61% have patient information leaflets. The majority of respondents were satisfied with their level of explanation regarding indications for treatment (69.3%) and the procedure (78.7%). Fifty-nine percent felt patients understood alternative treatment options. Only 37.8% of radiologists document possible complications in the patient's chart. Comments from respondents indicated that there is insufficient time for radiologists to obtain consent in all patients. Suggestions to improve current local policies included developing the role of radiology nursing staff and the use of radiology outpatient clinics. CONCLUSIONS: More than 50% of respondents are unhappy with their policies for obtaining informed consent. Interventional societies have a role to play in advocating formal consent guidelines. PMID- 14753300 TI - Endovascular stent-graft applications in latrogenic vascular injuries. AB - PURPOSE: To report the results of covered stent applications in iatrogenic vascular injuries. METHODS: We report 17 patients (11 men, 6 women; age range 20 59 years, mean age 40 years) who underwent repair of different iatrogenic vascular lesions by means of endovascular covered stents. The patient population consisted of 8 femoral arteriovenous fistulae, 4 common femoral artery pseudoaneurysms, 1 subclavian artery pseudoaneurysm, 1 abdominal aortic aneurysm, 1 iliac artery perforation, 2 porto-biliary fistulae that developed during TIPS procedure. Balloon-expandable stent-grafts were used in all patients except one. Control studies were performed with angiography. RESULTS: Technical success was achieved in all 17 patients. The mean clinical follow-up period for all 17 patients was 8 months. There were no signs of stent migration or leaks in the control studies. Only one patient developed a hemodynamically insignificant stenosis at the proximal end of the stent. There have been no stent deformations or related complications during the follow-up period. CONCLUSION: Our short-term results suggest that endovascular treatment is a low-risk procedure and appears less invasive than surgery for the treatment of different types of iatrogenic vascular injuries. Intermediate and long-term results are not available. PMID- 14753301 TI - Percutaneous retrieval of chronic intravascular foreign bodies. AB - To evaluate the feasibility of intravascular retrieval of chronic foreign bodies, we retrospectively reviewed an 8 year experience (1993-2001) of percutaneous retrieval of chronically retained intravascular foreign bodies (n = 6). In 6 of 6 cases (4 catheter fragments, 2 guidewires), 5-90 days elapsed before retrieval via the femoral or internal jugular vein. Under fluoroscopy, we determined the foreign body's course, position and size. A guidewire was advanced through a multipurpose catheter to the foreign body. The multipurpose catheter was replaced with a gooseneck snare catheter and the snare advanced to grasp and remove the foreign body. Percutaneous retrieval was successful in all 6 cases. One patient experienced mild hemoptysis, which resolved within 24 hr of observation. No patient experienced long-term sequelae. Given the potential life-threatening complications from intravascular foreign bodies and the low complication rate from percutaneous retrieval, we recommend extraction of the foreign body even if it is asymptomatic in the chronic setting (> 24 hr). PMID- 14753302 TI - Endovascular therapeutic options for isolated iliac aneurysms with a working classification. AB - The purpose of this paper is to demonstrate a variety of stent-grafting and embolization techniques and describe a new classification for endovascular treatment of isolated iliac artery aneurysms. A total of 19 patients were treated for isolated iliac aneurysms. Depending on the proximal iliac neck and the uni /bilaterality of common iliac artery aneurysms (CIAA's) the patient may be treated by a tube (Type Ia) or a bifurcated stent-graft (Type Ib) in addition to internal iliac artery embolization. Neck anatomy is also critical in determining therapeutical options for internal iliac artery aneurysms (IIAA's). These are tube stent-grafting plus internal iliac branch embolization (Type IIa), coiling of afferent and efferent internal iliac vessels (Type IIb) and IIAA packing (Type IIc). The average length of stay for these procedures was 3.8 days. During the mean follow-up of 20.9 months, aneurysm size remained unchanged in all but 4 patients. Reinterventions were necessary in option Type Ib (3/8 pat.) and Type Ia (1/7 pat.) due to extender stent-graft migration (n = 2) or reperfusion leaks (n = 2). We conclude that Iliac artery aneurysms may be successfully and safely treated by a tailored approach using embolization or a combination of embolization and stent-grafting. Long-term CT imaging follow-up is necessary, particularly in patients treated with bifurcated stent-grafts (Type Ib). PMID- 14753303 TI - Endovascular repair of complex aortic aneurysms: intravascular ultrasound guidance with an intracardiac probe. AB - To assess the accuracy and efficacy of intravascular ultrasound guidance obtained by an intracardiac ultrasound probe during complex aortic endografting. Between November 1999 and July 2002, 19 patients (5 female, 14 male; mean age 73.5 +/- 2.1 years) underwent endovascular repair of thoracic (n = 10), complex abdominal (n = 6) and concomitant thoraco-abdominal (n = 3) aortic aneurysm. The most suitable size and configuration of the stent-graft were chosen on the basis of preoperative computed tomographic angiography (CTA) or magnetic resonance angiography (MRA). Intraoperative intravascular ultrasound imaging was obtained using a 9 Fr, 9 MHz intracardiac echocardiography (ICE) probe, 110 cm in length, inserted through a 10 Fr precurved long sheath. The endografts were deployed as planned by CTA or MRA. Before stent-graft deployment, the ICE probe allowed us to view the posterior aortic arch and descending thoraco-abdominal aorta without position-related artifacts, and to identify both sites of stent-graft positioning. After stent-graft deployment, the ICE probe allowed us to detect the need for additional modular components to internally reline the aorta in 11 patients, and to discover 2 incomplete graft expansions subsequently treated with adjunctive balloon angioplasty. In 1 patient, the ICE probe supported the decision that the patient was ineligible for the endovascular exclusion procedure. The ICE probe provides accurate information on the anatomy of the posterior aortic arch and thoracic and abdominal aortic aneurysms and a rapid identification of attachment sites and stent-graft pathology, allowing refinement and improvement of the endovascular strategy. PMID- 14753304 TI - Histologic comparison of vibrating guidewire with conventional guidewire technique in an experimental coronary in vivo model. AB - PURPOSE: To compare the damage caused by vibrating guidewire manipulation and conventional guidewire manipulation of soft coronary wires in normal sheep coronary arteries. METHODS: Using an intact sheep model the two methods of passing a coronary guidewire down a normal coronary artery under fluoroscopic screening control were studied. The resulting arterial damage caused by the two techniques was studied histologically. The severity of damage was scored from 1 (no damage) to 4 (severe damage) and expressed as: (a) percentage of damaged sections, (b) mean damage score per section and (c) percentage of sections suffering the most severe degree of damage (scores 3 and 4). RESULTS: One hundred and sixty-eight sections were studied. The percentage of damaged sections was lower in the vibrating guidewire group (p = 0.004). The mean damage score and the percentage of sections with a damage score of 3 or 4 were smaller in the vibrating guidewire group than in the conventional guidewire manipulation group (p = 0.001 and p = 0.009, respectively). CONCLUSIONS: Both methods of guidewire manipulation cause identifiable vascular damage. The extent and severity of damage appear greater when the guidewire is manipulated manually. PMID- 14753305 TI - Comparison of small intestinal submucosa-covered and noncovered nitinol stents with PTFE endografts in injured ovine femoral arteries: a pilot study. AB - The purpose of this study was to compare performance of small intestinal submucosa (SIS)-covered endografts (SCEs) to polytetra-fluoroethylene (PTFE) covered endografts (PCEs) and to bare nitinol stents (BSs) in injured sheep femoral artery (FA). Bare Zilver 6 mm X 40 mm nitinol stents (n = 6), Zilver stents covered with SIS (n = 6), and Palmaz stents 6 mm x 37 mm covered with PTFE (n = 6) were implanted in the balloon-injured FAs of nine female sheep. Follow-up arteriograms were obtained before animal sacrifice at 1, 3 and 6 months, with three animals at each time point. The FAs with the implanted device were explanted for histologic studies and morphologic measurements. Stent implantation was technically successful in all sheep. All BS and SCEs were patent at each time point. Five BSs and five SCEs exhibited formation of progressive eccentric intimal hyperplasia (IH) that was more advanced in SCE at 6 months. Cross sectional area narrowing averaged 60% for BSs and 67% for SCEs. One BS, one SCE and two patent PCEs exhibited mild-to-moderate formation of concentric IH. Four PCS occluded one at 1 month, two at 3 months and one at 6 months. Performance of the devices placed into sheep FAs depended on their relation to the curving peri articular portion of the FA during extremity flexion. BSs and SCEs placed in this portion exhibited progressive growth of eccentric IH while PCEs placed in this portion occluded. PMID- 14753306 TI - Stent graft in managing juxta-renal aortoiliac occlusion. AB - Endovascular procedures are frequently used as an alternative to surgical bypass in aortic and iliac occlusion. Stents have revolutionized the scope of such endovascular procedures, but there are few reports of stents or stent grafts in occlusive juxta-renal aortic occlusion. We present a case where such occlusion was managed by use of a stent graft with successful outcome. PMID- 14753307 TI - Acute duodenal obstruction after percutaneous placement of metallic biliary stents: peroral treatment with enteral stents. AB - Three patients with malignant biliary obstruction were treated with placement of metallic biliary stents. Two patients had known partial duodenal stenosis but had no symptoms of gastrointestinal obstruction. The patients developed symptomatic duodenal obstruction early after biliary metallic stent placement. The symptomatic duodenal obstructions were successfully treated with peroral placement of duodenal stents, which obviated the need for surgical intervention. PMID- 14753308 TI - Successful treatment of isolated spontaneous superior mesenteric artery dissection with stent placement. AB - Isolated dissection of superior mesenteric artery is a rare condition and is usually treated surgically. We treated a patient with severe abdominal pain who was angiographically confirmed to have superior mesenteric artery thrombosis associated with isolated spontaneous dissection. He was treated initially by thrombolysis and oral anticoagulation, but recurrent symptoms developed with radiologic evidence of disease progression. We performed superior mesenteric artery stenting and recovery was uneventful. PMID- 14753310 TI - The use of direct thrombin injection to treat a type II endoleak following endovascular repair of abdominal aortic aneurysm. AB - This report describes the use of thrombin to treat a type II endoleak which was causing continued abdominal aortic aneurysm expansion in a patient who had undergone endovascular repair. A small quantity of thrombin was injected into the leak by a percutaneous approach directly into the aneurysm sac using color doppler ultrasound. The procedure was successful and required only a few minutes to perform. We believe this procedure is an alternative to some of the more complex and technically challenging means of treating this lesion. PMID- 14753309 TI - Transcatheter arterial embolization therapy for a hypoplastic pelvic kidney with a single vaginal ectopic ureter to control incontinence: the usefulness of three dimensional CT angiography using multidetector-row helical CT. AB - A girl with continuous urinary incontinence was successfully treated by angiographic embolization of a hypoplastic pelvic kidney with a single unilateral vaginal ectopic opening of the ureter. For this intervention, CT angiography was useful for detecting the corresponding renal artery of the hypoplastic kidney. PMID- 14753311 TI - Ruptured mycotic pulmonary artery pseudoaneurysm in an infant: transcatheter embolization and CT assessment. AB - Mycotic pseudoaneurysim of the pulmonary artery that ruptures during necrotizing pneumonia is a rare entity that is often fatal. Traditionally it has been treated with open thoracotomy and resection of both the aneurysm and the lobe in which the aneurysm is located. In this report, we describe the radiological findings and transcatheter coil embolization of a mycotic pulmonary pseudoaneurysm in a 6 month-old female infant. We also describe the subsequent morphologic changes observed on follow-up computed tomography after 9 months of embolization. PMID- 14753312 TI - Accessory meningeal arterial supply to the posterior nasal cavity: another reason for failed endovascular treatment of epistaxis. AB - A patient with intractable posterior epistaxis was treated with embolization of the ipsilateral sphenopalatine and facial arteries and contralateral sphenopalatine artery. She continued to bleed despite a seemingly adequate embolization procedure. A second angiogram revealed a significant collateral blood supply to the posterior nasal cavity from the accessory meningeal artery not identified during the first procedure. This was then embolized with no further epistaxis encountered. This case demonstrates yet another collateral arterial pathway that might account for a failed embolization. PMID- 14753313 TI - Temporary and permanent inferior vena cava filter combination in a young patient: to implant or not to implant? AB - The decision to implant vena cava filters, either temporary or permanent, is difficult in young patients. We present the case of a young man with pulmonary embolism in whom temporary and permanent inferior vena cava filters were implanted. The decision process is discussed in relation to the current literature. PMID- 14753314 TI - Fatal aortic rupture complicating stent plasty in a case of aortoarteritis. AB - Endovascular stenting has emerged as an effective alternative for unsuccessful angioplasty of the aorta in aortoarteritis. This is a single case report of fatal aortic rupture following balloon angioplasty of post-patch aortoplasty restenosis in aortoarteritis. We report a fatal aortic rupture during angioplasty of the primarily stented stenotic segment of the aorta in a case of aortoarteritis. PMID- 14753315 TI - Esophageal and bronchial perforations after thoracic aortic aneurysm replacement: successful repair with covered metallic stents. AB - Esophageal and bronchial perforations are rare but potentially fatal complications of descending thoracic aortic aneurysm replacement. This report presents a 67-year-old man with both esophageal and bronchial perforations that occurred after descending thoracic aortic aneurysm replacement. Surgical repair was performed, but the lesions perforated again. Two covered metallic stent prostheses introduced into both the esophagus and left main bronchus led to the improvement of mediastinitis by sealing the perforations. To our knowledge, this is the first report describing successful treatment for esophageal and bronchial perforations using covered metallic stents. Placement of covered metallic stents can be an option for the treatment of patients with esophagorespiratory tract perforations, especially those who are in critical condition. PMID- 14753316 TI - Delayed union of a sacral fracture: percutaneous navigated autologous cancellous bone grafting and screw fixation. AB - Delayed or non-union of a sacral fracture is a serious clinical condition that may include chronic pain, sitting discomfort, gait disturbances, neurological problems, and inability to work. It is also a difficult reconstruction problem. Late correction of the deformity is technically more demanding than the primary treatment of acute pelvic injuries. Open reduction, internal fixation (ORIF), excision of scar tissue, and bone grafting often in a multi-step approach are considered to be the treatment of choice in delayed unions of the pelvic ring. This procedure implies the risk of neurological and vascular injuries, infection, repeated failure of union, incomplete correction of the deformity, and incomplete pain relief as the most important complications. We report a new approach for minimally invasive treatment of a delayed union of the sacrum without vertical displacement. A patient who suffered a Malgaigne fracture (Tile C1.3) was initially treated with closed reduction and percutaneous screw fixation (CRPF) of the posterior pelvic ring under CT navigation and plating of the anterior pelvic ring. Three months after surgery he presented with increasing hip pain caused by a delayed union of the sacral fracture. The lesion was successfully treated percutaneously in a single step procedure using CT navigation for drilling of the delayed union, autologous bone grafting, and screw fixation. PMID- 14753317 TI - A case of an upper gastrointestinal bleeding due to a ruptured dissection of a right aortic arch. AB - We report a case of severe upper gastrointestinal hemorrhage with a rare underlying cause. The patient was unconscious when he was admitted to the hospital. No chest radiogram was performed. Routine diagnostic measures, including endoscopy, failed to reveal the origin of the bleeding, which was believed to originate from the esophagus secondary to a peptic ulcer or varices. Exploratory laparotomy added no further information, but contrast-enhanced multislice computed tomography (MSCT) of the chest showed dextroposition of the widened aortic arch with a ruptured type-B dissection and a consecutive aorto esophageal fistula (AEF). The patient died on the day of admission. Noninvasive MSCT angiography gives rapid diagnostic information on patients with occult upper gastrointestinal bleeding and should be considered before more invasive conventional angiography or surgery. PMID- 14753318 TI - Cognitive predictors of satisfaction with hospital inpatient service encounters among the elderly: a matter of trust. AB - Given the burgeoning growth of the elderly population, ensuring the satisfaction of senior consumers of health services is worthy of heightened attention by healthcare administrators both on economic and social grounds. By examining inpatient satisfaction among the elderly, we have focused our attention on an important and understudied segment of health consumers within a dominant service delivery context. Moreover, we have supplemented traditional hospital service satisfaction indicators by including a variety of rarely used psychological variables as cognitive predictors of service assessments. This study reinforces the empirical connection between trust in one's physician and elderly inpatient satisfaction at an increased level. The results suggest that increased levels of trust is positively associated with overall satisfaction of elderly inpatients as well as their satisfaction with several subdimensions of service that represent specific attributes of the hospital inpatient encounter. PMID- 14753319 TI - Trends in rural healthcare delivery in the United States, 1990-1999. AB - This article describes the results of a literature search of pertinent professional literature written on issues important to rural healthcare delivery in the United States. The purpose of the literature review was to provide journal articles to practicing rural healthcare leaders who, because of time and economic constraints, claimed they were unable to research their own relevant journal articles. The authoring team is composed of individuals who work as full-time faculty members at institutions that offer courses in health administration. Because the nature of their professional work and research dictates that they focus on areas of rural health, the authoring team offered to share their findings with rural administrators as a means to assist them. The intent was to form a working collaboration between health services management professionals in academia, representatives of the state's healthcare organizational affiliate associations, and professionals practicing in the industry. This ten-year collaboration resulted in a unique wedding of resources that helped both in the delivery of rural healthcare services in a western state and in the advancement of the field of health services administration through original research. Thus, the product of this literature search is a comprehensive study of the trends in rural healthcare delivery. The variety of topics discussed were gleaned from over 500 articles found in and summarized from 70 professional healthcare journals published between 1990 and 1999. PMID- 14753320 TI - Relationships between hospital-centered and multihospital-centered factors and perceived effectiveness: a canonical study of nonprofit hospitals. AB - This article reports on the results of a survey which investigated the nature of relationships between hospital and multihospital organization-centered factors and background characteristics, and multihospital organization effectiveness. Canonical correlation is employed in analyzing the data. Results and their implications are discussed. PMID- 14753321 TI - Magnet status as a competitive strategy of hospital organizations: marketing a culture of excellence in nursing services. AB - With issues of patient safety, the nursing shortage, and managed care fiscal constraints, hospital organizations can strategically capture market share, while insuring best care practices, if they adopt the "Magnet Status" accreditation model. This quality indicator signifies to the consumer a culture of excellence in nursing services and fulfills the priority of customer satisfaction as a marketing strategy objective. PMID- 14753322 TI - Financial statistics for public health dispensary decisions in Nigeria: insights on standard presentation typologies. AB - Public health dispensaries in Nigeria in recent times have demonstrated the poise to boost corporate productivity in the new millennium and to drive the nation closer to concretising the lofty goal of health-for-all. This is very pronounced considering the face-lift giving to the physical environment, increase in the recruitment and development of professionals, and upward review of financial subventions. However, there is little or no emphasis on basic statistical appreciation/application which enhances the decision making ability of corporate executives. This study used the responses from 120 senior public health officials in Nigeria and analyzed them with chi-square statistical technique. The results established low statistical aptitude, inadequate statistical training programmes, little/no emphasis on statistical literacy compared to computer literacy, amongst others. Consequently, it was recommended that these lapses be promptly addressed to enhance official executive performance in the establishments. Basic statistical data presentation typologies have been articulated in this study to serve as first-aid instructions to the target group, as they represent the contributions of eminent scholars in this area of intellectualism. PMID- 14753323 TI - Hospital marketing. AB - This article looks at a prescribed academic framework for various criteria that serve as a checklist for marketing performance that can be applied to hospital marketing organizations. These guidelines are drawn from some of Dr. Noel Capon of Columbia University's book Marketing Management in the 21st Century and applied to actual practices of hospital marketing organizations. In many ways this checklist can act as a "marketing" balanced scorecard to verify performance effectiveness and develop opportunities for innovation. PMID- 14753324 TI - Pfizer and its competitive marketing challenges. AB - Pfizer has been the preeminent global pharmaceutical company in recent years and much of their success can be attributed to their marketing responsiveness and innovation. Pfizer is committed to patients, the community and quality product development. As a model in marketing effectiveness they also show how to lead internal resources efficiently to maximize market place opportunities. PMID- 14753325 TI - Quantitative measurement of marginal disintegration of ceramic inlays. AB - The objectives of this study include establishing a method for quantitative measurement of marginal change in ceramic inlays and clarifying their marginal disintegration in vivo. An accurate CCD optical laser scanner system was used for morphological measurement of the marginal change of ceramic inlays. The accuracy of the CCD measurement was assessed by comparing it with microscopic measurement. Replicas of 15 premolars restored with Class II ceramic inlays at the time of placement and eight years after restoration were used for morphological measurement by means of the CCD laser scanner system. Occlusal surfaces of the restored teeth were scanned and cross-sections of marginal areas were computed with software. Marginal change was defined as the area enclosed by two profiles obtained by superimposing two cross-sections of the same location at two different times and expressing the maximum depth and mean area of the area enclosed. The accuracy of this method of measurement was 4.3 +/- 3.2 microm in distance and 2.0 +/- 0.6% in area. Quantitative marginal changes for the eight year period were 10 x 10 microm in depth and 50 x 10(3) microm2 in area at the functional cusp area and 7 x 10 microm in depth and 28 x 10(3) microm2 in area at the non-functional cusp area. Marginal disintegration at the functional cusp area was significantly greater than at the non-functional cusp area (Wilcoxon signed ranks test, p < 0.05). This study constitutes a quantitative measurement of in vivo deterioration in marginal adaptation of ceramic inlays and indicates that occlusal force may accelerate marginal disintegration. PMID- 14753326 TI - Comparative depths of cure among various curing light types and methods. AB - This study evaluated the depth of cure associated with commercial LEDs (light emitting diodes) (Elipar FreeLight [FL], 3M-ESPE; GC e-Light [EL], GC), high intensity (Elipar TriLight [TL], 3M-ESPE) and very high intensity (Astralis 10 [AS], Ivoclar Vivadent) Quartz Tungsten Halogen (QTH) curing lights. Depth of cure of the various lights/curing modes were compared to a conventional QTH light (Max [Mx], Dentsply-Caulk). Ten exposure regimens were investigated: FL1 - 400 mW/cm2 [40 seconds]; FL2 - 0-400 mW/cm2 [12 seconds] --> 400 mW/cm2 [28 seconds]; EL1 - 750 mW/cm2 [10 pulses x 2 seconds], EL2 - 350 mW/cm2 [40 seconds]; EL3 - 600 mW/cm2 [20 seconds]; EL4 - 0 - 600 mW/cm2 [20 seconds] --> 600 mW/cm2 [20 seconds]; TL1 - 800 mW/cm2 [40 seconds]; TL2 - 100- 800 mW/cm2 [15 seconds] --> 800 mW/cm2 [25 seconds]; AS1 - 1200 mW/cm2 [10 seconds]; MX - 400 mW/cm2 [40 seconds]. Depth of cure was determined by penetration, scraping and microhardness techniques. The results were analyzed using one-way ANOVA/Scheffe's post-hoc test and Pearson's correlation at significance level 0.05 and 0.01, respectively. All light curing regimens met the ISO depth of cure requirement of 1.5 mm with the exception of EL1-EL3 with the microhardness technique. Curing with most modes of EL resulted in significantly lower depths of cure than the control [MX]. No significant difference in depth of cure was observed among the control and the two modes of FL. Curing with TL1 resulted in significantly greater depth of cure compared to MX with all testing techniques. No significant difference in depth of cure was observed between the control and AS1 for all testing techniques except for the penetration technique. The depth of composite cure is light unit and exposure mode dependent. Scraping and penetration techniques were found to correlate well but tend to overestimate depth of cure compared to microhardness. PMID- 14753327 TI - Simultaneous release of fluoride and aluminum from dental materials in various immersion media. AB - Since fluoride (F-) and aluminum (Al3+) present anticariogenic properties and F- release from ionomeric materials depends on the media used in the evaluation, this study tested the hypothesis whether release of Al3+ also depends on the testing media. The simultaneous release of F- and Al3+ was assessed over 15 days in three media: (i) distilled and deionized water (H2O), (ii) artificial saliva (AS) and (iii) solutions simulating a cariogenic challenge (pH-cycling in demineralizing and remineralizing solutions, De-Re-). Six cylindrical samples of each tested material (Ketac-Fil, Vitremer, Fuji Ortho LC and F 2000) were prepared and suspended individually in 1.0 mL of each solution. All solutions were changed daily. F- and Al3+ were determined by ion-selective electrode and atomic absorption, respectively. ANOVA showed statistical significance for the interaction of material, time, media (p < 0.05), either for F- or Al3+ release. The resin-modified glass ionomer Vitremer released the highest amount of F- and Al3+ in De-Re- solutions compared to the other materials (p < 0.05); differences among the materials in H2O and AS were statistically consistent. The data suggest that the media used to evaluate the simultaneous release of F- and Al3+ should be taken into account when the anticariogenic potential of different dental materials is assessed. PMID- 14753328 TI - Biocompatibility of a flowable composite bonded with a self-etching adhesive compared with a glass lonomer cement and a high copper amalgam. AB - This study evaluated the pulpal response and in-vivo microleakage of a flowable composite bonded with a self-etching adhesive and compared the results with a glass ionomer cement and amalgam. Cervical cavities were prepared in monkey teeth. The teeth were randomly divided into three groups. A self-etching primer system (Imperva FluoroBond, Shofu) was applied to the teeth in one of the experimental groups, and the cavities were filled with a flowable composite (SI BF-2001-LF, Shofu). In the other groups, a glass ionomer cement (Fuji II, GC) or amalgam (Dispersalloy, Johnson & Johnson) filled the cavity. The teeth were then extracted after 3, 30 and 90 days, fixed in 10% buffered formalin solution and prepared according to routine histological techniques. Five micrometer sections were stained with hematoxylin and eosin or Brown and Brenn gram stain for bacterial observation. No serious inflammatory reaction of the pulp, such as necrosis or abscess formation, was observed in any of the experimental groups. Slight inflammatory cell infiltration was the main initial reaction, while deposition of reparative dentin was the major long-term reaction in all groups. No bacterial penetration along the cavity walls was detected in the flowable composite or glass ionomer cement except for one case at 30 days in the glass ionomer cement. The flowable composite bonded with self-etching adhesive showed an acceptable biological com- patibility to monkey pulp. The in vivo sealing ability of the flowable composite in combination with the self-etching adhesive was considered comparable to glass ionomer cement. Amalgam restorations without adhesive liners showed slight bacterial penetration along the cavity wall. PMID- 14753329 TI - Fracture resistance of compomer and composite restoratives. AB - This study evaluated and compared the fracture toughness of compomers and composites. Three compomer (Compoglass F [CG], Vivadent; F2000 [FT], 3M-ESPE; Dyract Posterior [DP], Dentsply) and three composite (Tetric Ceram [TC], Vivadent; Z250 [ZT], 3M-ESPE; Esthet X [EX], Dentsply) restoratives were selected for the study. Single-edged notched specimens (25 x 2 x 2 mm) were fabricated according to manufacturers' instructions and conditioned in distilled water at 37 degrees C for one week prior to testing. Seven specimens were made for each material. The specimens were loaded to failure using an Instron microtester with a crosshead speed of 0.5 mm/minute. Data were subjected to ANOVA/Scheffe's test and Independent Samples T-test at significance level 0.05. The mean fracture toughness (K(IC)) ranged from 0.97 to 1.23 MPam 1/2 for compomers and 1.75 to 1.92 MPam 1/2 for composites. The fracture toughness of compomers was significantly lower than their composite counterparts. No significant difference in K(IC) values was observed among the different composites. When the compomers were compared, FT had significantly higher fracture toughness than DP and CG. In view of their poorer resistance to crack propagation, compomers are not recommended for use in stress-bearing areas. PMID- 14753330 TI - Microleakage and marginal hybrid layer formation of compomer restorations. AB - The tested hypotheses of this study were that dentin enamel bonding agents (DBAs) proposed for compomers create a hybrid layer (HL) that seals the margins of Class V restorations and HL is free from voids or gaps on both enamel and dentin margins. For purposes of this study, Class V restorations (n = 70) were made in vitro at the CEJ in extracted third molars. Different systems (bonding agent + compomer) were selected. After finishing with discs, each margin was polished with diamond polishing paste for one minute, treated with a 2.5% NaOCl gel for 10 seconds and washed with deionized water to remove polishing debris and non infiltrated collagen. All restorations were immersed in dye solution for 24 hours, then inspected along the margins. SEM analysis was used to evaluate the morphology of the marginal HL and microleakage tests to evaluate their ability to seal the margins of restorations. Marginal leakage was observed along the dentin and enamel margin. A thin marginal HL (0.5-1.2 microm) was detected only along the dentin margin of several bonding systems but not along the enamel margin. Porosities and gaps were detected along margins when no HL was observed. The results demonstrated that the tested bonding agents for compomers produced a thin marginal hybrid layer, especially along the dentin margin. Microleakage had a relationship with the morphology (gap, porosities and thickness) of this hybrid smear layer. In conclusion, the DBAs tested specifically developed for compomers did not ensure an intimate interfacial adaptation, because microleakage was detected along the enamel and dentin interfaces and the marginal hybrid layer was only partially homogeneous. PMID- 14753331 TI - Thermal and mechanical load cycling on microleakage and shear bond strength to dentin. AB - This study evaluated the influence of mechanical and thermal cycling on microleakage at the cervical margins of proximal slot restorations and shear bond strength on flat dentin surfaces. Microleakage Evaluation: One hundred and twenty slot cavity restorations were performed on bovine incisors. The restorations were randomly divided into four groups (n = 30): control (no thermal and mechanical load cycling), thermal cycling (2,000 cycles, 5 degrees C-55 degrees C), mechanical load cycling (50,000-80N) and thermal and load cycling (2,000 5 degrees C-55 degrees C/50,000-80N). The specimens were sealed with acid resistant varnish, leaving a 1-mm window around the cervical margin interface. To detect marginal leakage, a 2% methylene blue buffered solution was used for four hours. The specimens were sectioned longitudinally and qualitatively evaluated by stereomicroscopy (45x) following a ranked score for the dentin cervical margin. The data were analyzed by Kruskal-Wallis test (alpha = 0.05). Shear Bond Strength Evaluation (SBS): Eighty bovine incisors were embedded and polished to obtain a flat standard surface on dentin. The surfaces were restored with Single Bond adhesive system and a resin composite subsequently inserted in a bipartite Teflon matrix. The specimens were randomly divided into the four groups (n = 20) described above for microleakage. Shear bond strengths were measured in a universal testing machine with a crosshead speed of 0.5 mm/minute. The data were analyzed by one way ANOVA test (alpha = 0.05). No statistically significant influence of thermocycling, mechanical load cycling or the combination was observed for both microleakage and shear bond strength. PMID- 14753332 TI - Support of occlusal enamel provided by bonded restorations. AB - In this in vitro study, the resistance to fracture of occlusal enamel supported by a bonded tooth-colored restorative material was compared to unsupported enamel and enamel supported by sound dentin. Eighty extracted human lower molars were sorted into five groups of 16 teeth each. Lingual cusps were removed. In Groups 2 5, dentin was removed from the facial cusps, leaving a shell of enamel. In Group 1, dentin was not removed. Group 2 remained unrestored. The groups in which a restorative material was inserted to replace missing dentin were as follows: composite (Scotchbond MP/Filtek Z250 [A2] in capsules, 3M); Group 4-resin modified glass-ionomer (Fuji II LC [A2] in capsules, GC); Group 5-conventional glass ionomer (Fuji IX [A2] in capsules, GC). Specimens were thermocycled (1500 cycles, 6 degrees-60 degrees C, dwell 30 seconds), then mounted in die stone with lingual inclines of facial cusps approximately horizontal. The cusp ridges of the lingual inclines were flattened slightly using a horizontally mounted separating disk. Specimens were loaded evenly on flattened inclines in an Instron with a flat rectangular rod at a crosshead speed of 5 mm/minute. Data analysis was with one-way ANOVA and Student-Newman-Keul's test (F = 50.30, p < 0.0001). The bonded restorations provided significantly less enamel support than natural dentin and significantly more than when the enamel was left without support by dentin or a restorative material. There was no difference in support provided by the three restorative materials. PMID- 14753333 TI - Effect of different application and polymerization techniques on the microleakage of proximal resin composite restorations in vitro. AB - This in vitro study investigated cervical microleakage of proximal resin composite restorations placed with three application and polymerization techniques. Uniform mesio-occlusal-distal (MOD) preparations featuring cervical margins above (mesial) and below (distal) the CEJ suitable for restoration with resin composite were copy milled into 33 recently extracted permanent molars. The teeth were divided randomly into three groups of 11 teeth and restored using a conventional incremental technique (Group A) and two novel curing devices (Groups B and C). After 24 hours, a dye penetration test was used to assess microleakage. Conventional placement in preparations with cervical margins in enamel had significantly lower interfacial leakage scores than those recorded for placement in preparations with margins in cementum regardless of the technique used to place the restorative material. Use of the two novel curing devices conferred no advantage in reducing microleakage irrespective of where preparation margins were placed. PMID- 14753334 TI - Effect of surface treatment of prefabricated posts on bonding of resin cement. AB - This in vitro study evaluated the effect of various surface treatments of prefabricated posts of titanium alloy (ParaPost XH), glass fiber (ParaPost Fiber White) and zirconia (Cerapost) on the bonding of two resin cements: ParaPost Cement and Panavia F by a diametral tensile strength (DTS) test. The posts received surface treatments in three categories: 1) roughening by sandblasting and hydrofluoric acid etching; 2) application of primer by coating with Alloy Primer, Metalprimer II and Silane and 3) a combination treatment in the form of roughening (sandblasting or etching) supplemented by the application of a primer or in the form of the Cojet system. After surface treatment, the post was embedded in a cylinder of resin cement (diameter = 4.0 mm, height = 4.0 mm). The surface-treated post was centered in the resin cement-filled mold with the aid of fixation apparatus. Fifteen minutes from the start of mixing the resin cement, the specimen was freed from the mold and stored in water at 37 degrees C for seven days. Following water storage, the specimen was wet-ground to a final length of approximately 3 mm. The DTS of specimens was determined in a Universal Testing Machine. The bonding of resin cement to titanium alloy posts was increased by several surface treatments of the post. However, coating with primers as sole treatment had no effect on bonding. With the DTS method applied, none of the surface treatments had an effect on the bonding to glass fiber posts. The bonding of both resin cements to zirconia posts was improved by Cojet treatment, while sandblasting, followed by silane application, improved bonding of Panavia F. PMID- 14753335 TI - Tensile bond strength of composite to air-abraded dentin. AB - This study evaluated the influence of air abrasive treatment of dentin surfaces on the tensile bond strength between dentin and two different composite-adhesive systems Multi-Purpose/Z100 and OptiBond FL/Herculite XR). The crowns of 200 maxillary central incisors were embedded in resin and then ground to expose a dentin surface 5 mm in diameter. The surfaces were etched or abraded by using a KCP 1000 device with different treatment conditions. Adhesive systems were applied according to the manufacturer's instructions and composite cylinders were bonded to the conditioned dentinal surface using a split mold. Tensile bond strength values and failure modes were then determined. Tensile bond strength values of the acid-etched dentin-composite-interface were significantly higher than for the interface between air-abraded dentin and composite, independent of the composite-adhesive-system used. The light microscopic evaluation showed mainly adhesive and combined adhesive-cohesive fractures. Significantly more adhesive fractures could be observed between abraded dentin and composite than between etched dentin and composite. PMID- 14753336 TI - Effect of an "all-in-one" adhesive on pulp blood vessels: a vitalmicroscopic study of rat's teeth. AB - The newly developed self-etching, self-priming all-in-one adhesives are appealing to clinicians because they are simple and efficient to use. These single application bonding systems contain chemically active compounds that can alter pulpal blood circulation when applied to deep dentin surfaces. Since adequate microcirculation and oxygenation are the basic requirements for tissue survival, the aim of this study was to investigate the immediate vascular effect of a new self-etching adhesive, Prompt L-Pop/composite and compomer version/(test group). The technique of vitalmicroscopy was used to record the changes in vessel diameter of the first lower incisor of 20 (10-10 in each group) male Sprague Dawley rats (weighing 315 +/- 74/SE/g) prior to, and at 5, 15, 30 and 60 minutes after the investigated materials were applied to dentin. The application of saline served as the untreated control. The systemic arterial pressure remained unchanged throughout the experiments both in the control (110 +/- 8 mmHg) and in test animals (114 +/- 4 mmHg). In control rats, the diameter of the vessels was stable during the experiment. In the presence of Prompt L-Pop, the diameters decreased significantly during the experimental period (5 minutes: -11.15 +/- 5.03%; 15 minutes: -14.66 +/- 7.71%; 30 minutes: -13.35 +/- 5.79%; 60 minutes: 11.82 +/- 5.63% p < 0.05 in each cases). In this group, stasis developed in pulpal circulation was 1 out of 10 rats. The results from the rat model used in this study suggest that Prompt L-Pop may result in compromised pulpal microcirculation. PMID- 14753337 TI - Finishing tooth-colored restorations in vitro: an index of surface alteration and finish-line destruction. AB - Many studies have evaluated the surface characteristics of finishing and polishing instruments on different restorative materials using two- and three dimensional models based on mechanical and optical techniques. However, only limited data are available regarding the problem of marginal causing detectable surface alterations such as scratches or grooves may also cause marginal damage. This study aimed to correlate the smooth-surface polishing efficacy of different instruments with their potential for destructive effects on restoration margins and enamel finish lines. An index was created that will help to evaluate future polishing instruments and select suitable ones for different clinical situations. A planar inlay system with a 100 microm wide defined gap was simulated in vitro. Pre-fabricated ceramic (n = 40) and composite blocks (n = 40) were connected to bovine enamel without luting material. After standardized pre-polishing, mean surface roughness and marginal quality were assessed using a profilometer and scanning electron microscopy (SEM). Enamel and restorative surfaces were colored, and subsequently prepared using one of 10 different finishing and polishing instruments. Four specimens per instrument and material were evaluated, resulting in eight interfaces for each test group. Surface roughness (Ra) and marginal quality (expressed as the percentage fracture-free margin) were measured and compared statistically using unpaired t-tests and two-way ANOVA, respectively. The level of significance was set at 0.05 Eight-micrometer diamond burs and 40 fluted tungsten carbide finishers produced smoother surfaces and less finishing line destructions than the other instruments under evaluation. The index values developed will prove helpful in evaluating and selecting appropriate instruments. PMID- 14753338 TI - Surface finish of resin-modified and highly viscous glass ionomer cements produced by new one-step systems. AB - This study investigated the surface finish of resin-modified (Fuji II LC, GC) and highly viscous (Fuji IX GP Fast, GC) glass ionomer cements after treatment with three one-step finishing/polishing systems (One-Gloss [OG], Shofu; Pogo [PG], Dentsply; Sof-Lex Brush [SB], 3M-ESPE). The surface roughness obtained was compared to that using a matrix strip [MS], a two-step rubber abrasive (CompoSite [CS], Shofu) and a graded abrasive disk (Super Snap [SS], Shofu) system. Eight specimens (3-mm long x 3-mm wide x 2-mm deep) of each material were made for the various treatment groups. With the exception of the MS group, all groups were roughened with 320 grit grinding paper using a lapping device prior to finishing/polishing with the different systems. The mean surface roughness (microm) was measured with a profilometer. Data was subjected to ANOVA/Scheffe's tests at significance level 0.05. Mean Ra ranged from 0.13 to 1.04 microm for Fuji II LC and 0.14 to 0.81 for Fuji IX GP. For both materials, the smoothest surface was obtained with MS and the roughest with OG. Depending on the materials, the surface finish produced by PG and SB was superior or comparable to that obtained with CS and SS. The effectiveness of one-step systems, when used to finish/polish resin-modified and highly viscous glass ionomer cements, is product dependent. PMID- 14753339 TI - Composite cure and pulp-cell cytotoxicity associated with LED curing lights. AB - This study compared the cure and pulp-cell cytotoxicity of composites polymerized with light-emitting diode (LED) and halogen-based light curing units. A mini filled resin composite (Tetric Ceram, Vivadent), two LED (E-light [EL], GC and Freelight [FL], 3M-ESPE), a conventional halogen (Max [MX], Dentsply) and a high intensity halogen light (Astralis 10 [AS], Vivadent) were evaluated. Cure associated with the different lights was determined by measuring the top and bottom surface hardness (KHN; n = 5) of 2-mm thick specimens using a digital microhardness tester (load = 500 gf; dwell time = 15 seconds). Pulp-cell cytotoxicity was assessed using a direct contact method involving incisor tooth slices dissected from 28-day old Wistar rats maintained in Dulbecco's Modified Eagle's Medium (DMEM) and 1% agarose. The bottom surfaces of the cured composite specimens (7-mm diameter and 2-mm deep) were placed in contact with the openings of each tooth slice. After incubation in 5% CO2 atmosphere at 37 degrees C for 48 hours, the tooth slices were fixed, demineralized and processed for histological examination. Pulp fibroblasts and odontoblasts were counted histomorphometrically at 400x magnification within a 1500 microm2 area using a computerized micro imaging system. Eighteen readings were obtained for each curing light. Data was subjected to ANOVA/Scheffe's test and Pearson's correlation at significance level 0.05 and 0.01, respectively. At the top surfaces, the cure with AS was significantly greater than the other curing lights, with MX and FL being significantly greater than EL. At the bottom surfaces, MX, AS and FL had significantly better cure than EL. Specimens cured with MX were less cytotoxic than those polymerized with other curing lights. Specimens cured with AS and EL were significantly less cytotoxic than FL. Composite cure and cytotoxicity associated with LED lights is device dependent. Composite cure was not correlated to pulp-cell cytotoxicity. The response of pulpal fibroblasts to unreacted/leached components of composites differs somewhat from odontoblasts. PMID- 14753340 TI - Comparison of surface finish of new aesthetic restorative materials. AB - This study compared the surface finish of eight different types of aesthetic restorative materials. The materials included resin-modified (Fuji II LC [FL], GC) and highly viscous (Fuji IX GP Fast [FN], GC) glass ionomer cements, a compomer (F2000 [FT], 3M-ESPE), minifilled (Z100 [ZO], 3M-ESPE) and microfilled (A110 [AO], 3M-ESPE) composites and materials based on recently introduced ormocer (Admira [AM], Voco), nanomer (Filtek Supreme Translucent [FST], 3M-ESPE) and nanocluster technology (Filtek Supreme [FS], 3M-ESPE). Sixteen specimens (3 mm long x 3-mm wide x 2-mm deep) of each material were divided into two equal groups. Specimens in Group 1 received no further treatment after polymerization against a matrix strip, while the specimens in Group 2 were roughened with 320 grit grinding paper using a lapping device and were finished/polished with a graded abrasive disk system (Super-Snap, Shofu). The mean roughness (Ra, microm) of materials was determined using a surface profilometry. Data was analyzed by ANOVA/Scheffe's test at significance level 0.05. Mean Ra values ranged from 0.04 to 0.16 microm for Group 1 specimens and 0.15 to 0.68 microm for Group 2 specimens. Results of statistical analysis were as follows: Group 1-FS, FST, FL, FN, AM > FT, AO, ZO; Group 2-FN, FT, FL > AO, FS, ZO, AM, FST (> indicates significantly greater Ra values). For the finished/polished composite materials, Ra values observed for AM and FST were significantly lower than for AO and FS. The surface finish of glass ionomers and compomer was significantly poorer than composites. Composite materials based on ormocer and nanomer technology were significantly smoother than those based on microfillers and nanoclusters. PMID- 14753341 TI - Polymerization efficiency of curing lamps: a universal energy conversion relationship predictive of conversion of resin-based composite. AB - A universal energy-conversion relationship (ECRu) predictive of conversion of a resin-based composite (RBC) polymerized with any light source has been described. This relationship was derived from an energy conversion relationship for RBC polymerized with a tungsten-halogen lamp and the lamp's efficiency relative to a hypothetical standard lamp. The ECRu was then used to predict conversion throughout RBC polymerized with an LED lamp using the lamp's relative efficiency compared to the standard lamp. The universal energy scale has also been described as predictive of scrape-back lengths for this RBC family when polymerized with any light source. Despite a 31% greater relative efficiency, scrape-back lengths from RBC polymerized using the LED lamp were predicted to be only 6% greater than those polymerized with the tungsten-halogen lamp when RBC is polymerized on an equal energy basis. This result was experimentally verified. PMID- 14753342 TI - Custom-made resin post and core. AB - Resin posts are used more often than metal posts, as they are esthetically compatible and less damaging to remaining tooth structure. Many resin post systems are available, where a light transmitting post is used to build up the composite in layers. An easier and economical alternative method, a custom-made resin post, is described. PMID- 14753343 TI - Assessment of positional candidate genes myf5 and igf1 for growth on bovine chromosome 5 in commercial lines of Bos taurus. AB - Quantitative trait loci for growth traits in beef cattle have been previously reported and fine-mapped in three chromosomal regions of 0 to 30 cM, 55 to 70 cM, and 70 to 80 cM of bovine chromosome 5. In this study, we further examined the association between gene-specific single nucleotide polymorphisms (SNP) of two positional candidate genes, bovine myogenic factor 5 (myf5) and insulin-like growth factor-1 (igf1), in the QTL regions and the birth weight (BWT), preweaning average daily gain (PWADG), and average daily gain on feed (ADGF) in commercial lines of Bos taurus. The QTL regions for the growth traits identified using a haplotype association analysis, which included the gene-specific SNP markers for both genes in this study, were in agreement with previous studies. The gene specific SNP marker association analysis indicated that the SNP in myf5 had a significant additive effect on PWADG in the M1 line of Beefbooster Inc. (P < 0.10), and a significant additive effect (P < 0.05) and a significant dominance effect (P < 0.10) on ADGF in the M3 line of Beefbooster Inc. When the data from the two commercial lines were pooled, the SNP in myf5 showed a significant association with PWADG (P < 0.10) and with ADGF (P < 0.05). The association between the SNP and BWT, however, did not reach a significance level in the M1 line, the M3 line, or across the lines. For igf1, no significant association between the SNP and the growth traits was detected in either the M1 line or the M3 line, whereas there was only a significant dominance effect (P < 0.10) on BWT detected for the SNP in igfl when the data from the two commercial lines were pooled. These results suggest that myf5 is a strong candidate gene that influences PWADG and ADGF in beef cattle. The SNP of igf1 may not be a causative or close to the causative mutation that affects the three growth traits in the populations of beef cattle examined in this study. Other SNP of igf1 and myf5 or other genes in their respective chromosomal regions, however, should also be studied. PMID- 14753344 TI - Allelic polymorphism in the ovine DQA1 gene. AB - Variation in the ovine DQA1 gene was investigated by amplification of exon 2 using PCR, followed by single-strand conformational polymorphism (SSCP) analysis, cloning, and DNA sequencing. Fourteen novel SSCP patterns, representing 14 different sequences, were identified. Eight of these 14 sequences were identical to published DQA1 sequences from sheep, whereas the remaining six were novel but similar to the published DQA1 sequences from sheep and cattle. These six new sequences exhibited conserved region and variable region patterns similar to the published sheep DQA1 sequences, but were different than the published DQA2 sequences from sheep. All of these 14 putative sheep DQA1 sequences fulfilled the criteria used by the established bovine leukocyte antigens major histocompatibility complex nomenclature committee for assignment as new alleles. Comparison of the available DQA1 sequences from sheep and cattle revealed several clusters of ovine DQA1 sequences, and some sheep alleles were more similar to cattle alleles than other sheep alleles. The occurrence of trans-species polymorphism suggests the action of balancing selection at the DQA1 locus. Twenty four percent of the nucleotide positions showed variation within exon 2, and this variation seems to have arisen largely by point mutation and gene conversion. The nonsynonymous and synonymous substitution rates were similar in both the putative antigen-binding site codons and the putative nonantigen-binding site codons. The extensive polymorphism reported in this article is consistent with polymorphism reported at the bovine DQA1 locus. PMID- 14753345 TI - Evaluation of gene expression in pigs selected for enhanced reproduction using differential display PCR and human microarrays: I. Ovarian follicles. AB - Differential display PCR (ddPCR) and complementary DNA microarray analyses were used to evaluate gene expression differences in porcine ovarian follicles between a line of pigs selected for an index of ovulation rate and embryo survival (Line I) and its randomly selected control line (Line C). Follicles (4.0 to 7.0 mm) were dissected from ovaries of multiparous sows (n = 27) at either 2 or 4 d following PGF2alpha analog injection on d 12 to 14 of the estrous cycle. Using ddPCR, differentially expressed bands (n = 282) were excised from gels and 107 were sequenced, yielding 84 unique porcine follicle expressed sequence tags. Northern hybridization confirmed differential expression (between lines, days, or follicle sizes) for messenger RNA representing the calpain I light subunit, cytochrome C oxidase subunit III, cytochrome P450 aromatase, and cytochrome P450 side chain cleavage genes. For microarray analysis, two mRNA pools representing follicles (d 2; 4.50 to 4.75 mm) from Line I and Line C sows were hybridized to the Incyte UniGEM V1.0 human chip (approximately 7,000 gene probes). A second analysis was performed using mRNA from follicles (d 2; 4.50 to 5.00 mm) hybridized to the Incyte UniGEM V2.0 human chip (approximately 9,100 gene probes). A total of 33 and 21 genes were identified with significant expression differences using UniGEM V1.0 and V2.0, respectively (twofold or greater relative expression following adjustment for expression of control probes). However, there was little overlap between results of the two hybridizations. Expression differences between lines for two genes, follistatin and nuclear receptor subfamily 4, group A, member 1, were confirmed using Northern hybridization. These results demonstrate changes in follicular gene expression as the result of long-term selection for enhanced reproduction. These correlated responses may directly represent allelic variation utilized by selection (e.g., quantitative trait loci), or more likely, transcriptional changes in other genes that interact with reproductive QTL. This work represents one of the first applications of gene expression analysis to evaluate long-term selection response in livestock populations. PMID- 14753346 TI - Evaluation of gene expression in pigs selected for enhanced reproduction using differential display PCR: II. Anterior pituitary. AB - The objective of this study was to identify differentially expressed genes in the anterior pituitary (AP) of sows selected for enhanced reproductive phenotypes. Selection in the Index (I) line was based on an index of ovulation rate and embryo survival, whereas random selection was used in the Control (C) line. Average numbers of fully formed piglets at birth were 12.5 +/- 1.5 and 9.9 +/- 2.0 for Line I and C sows used in this study, respectively. In order to induce luteolysis and synchronize follicle development, sows were injected (i.m.) with 2 mL of prostaglandin F2alpha analog between d 12 and 14 of the estrous cycle. Tissue was harvested 2 d (d2) or 4 d (d4) after injection, resulting in four experimental groups: Cd2 (n = 6), Cd4 (n = 4), Id2 (n = 6), and Id4 (n = 7). Differential display PCR (ddPCR) was used to search for transcriptional changes between selection lines in the AP, using samples within line but pooled across days. Northern hybridization was used to confirm ddPCR results. For ddPCR, two pools were used from each line (C and I). Three genes were confirmed to be differentially expressed between Lines I and C: G-beta like protein, ferritin heavy-chain, and follicle stimulating hormone beta subunit, whereas many other expressed sequence tags were observed to be differentially expressed but still require confirmation. Our findings indicate that long-term selection to increase ovulation rate and decrease embryo mortality has altered transcriptional patterns in the anterior pituitary, most likely as correlated responses. PMID- 14753347 TI - The National Pork Producers Council Maternal Line National Genetic Evaluation Program: a comparison of six maternal genetic lines for female productivity measures over four parities. AB - Litter (n = 8,424) and female performance records were collected in two breed-to wean production units in order to evaluate genetic line differences for sow longevity and maternal performance over four parities. Lines evaluated were American Diamond Genetics, Danbred North America, Dekalb-Monsanto DK44, Dekalb Monsanto GPK347, Newsham Hybrids, and National Swine Registry. Females within a line were derived from a minimum of 65 sires, 197 dams (three dams per sire), and a maximum of three daughters per dam, except in the GPK347, which were produced using semen from 12 Nebraska Index boars mated with Dekalb-Monsanto Line 34 females. All lines expressed 100% maternal heterosis. Mixed model statistical procedures were used with fixed effects including genetic line, parity, production unit, and two-way interactions. Random effects included a contemporary week of production and female for repeated records. Lactation length (average 15 d) was included as a linear covariate where appropriate. In total, 3,599 females entered as early-weaned pigs, 3,283 entered the breeding herd, 2,592 farrowed at least a single litter, and 1,656 and completed four parities. Line (P < 0.001) and parity (P < 0.001) effects were observed for virtually all traits measured. Ranges of genetic line differences averaged across parities were 1.76 pigs for total born, 1.45 pigs born alive, and 0.31 stillborn pigs per litter. Ranges of line differences in total and live litter weight were 1.4 and 1.3 kg, respectively. Ranges among lines, within Parities 1 through 4, for litter size at weaning were 0.56, 1.08, 0.91, and 0.64 pigs per litter, respectively. Line differences for weight (33.8 kg) and backfat depth (6.4 mm) at farrowing, lactation feed intake (8.7 kg), weight loss (5.0 kg), and backfat loss (0.87 mm) were observed. Extended wean-to-estrus interval was related to variation in weight, feed intake, and backfat loss in all lines except the GPK347. The GPK347 females farrowed and weaned the largest number of pigs, ate less feed in lactation, and lost more backfat and weight during lactation, yet they had the largest litters and the shortest wean-to-estrus intervals. Line x parity interactions existed for many traits due to small rank changes, but in general, the high- and low-ranked lines did not change. Genetic line differences in reproductive efficiency through four parities exist and must be recognized when choosing a female line. PMID- 14753348 TI - Covariance functions and random regression models for cow weight in beef cattle. AB - Data from the first four cycles of the Germplasm Evaluation program at the U.S. Meat Animal Research Center were used to evaluate weights of Angus, Hereford, and F1 cows produced by crosses of 22 sire and 2 dam (Angus and Hereford) breeds. Four weights per year were available for cows from 2 through 8 yr of age (AY) with age in months (AM). Weights (n = 61,798) were analyzed with REML using covariance function-random regression models (CF-RRM), with regression on orthogonal (Legendre) polynomials of AM. Models included fixed regression on AM and effects of cow line, age in years, season of measurement, and their interactions; year of birth; and pregnancy-lactation codes. Random parts of the models fitted RRM coefficients for additive (a) and permanent environmental (c) effects. Estimates of CF were used to estimate covariances among all ages. Temporary environmental effects were modeled to account for heterogeneity of variance by AY. Quadratic fixed regression was sufficient to model population trajectory and was fitted in all analyses. Other models varied order of fit and rank of coefficients for a and c. A parsimonious model included linear and quartic regression coefficients for a and c, respectively. A reduced cubic order sufficed for c. Estimates of all variances increased with age. Estimates for older ages disagreed with estimates using traditional bivariate models. Plots of covariances for c were smooth for intermediate, but erratic for extreme ages. Heritability estimates ranged from 0.38 (36 mo) to 0.78 (94 mo), with fluctuations especially for extreme ages. Estimates of genetic correlations were high for most pairs of ages, with the lowest estimate (0.70) between extreme ages (19 and 103 mo). Results suggest that although cow weights do not fit a repeatability model with constant variances as well as CF-RRM, a repeatability model might be an acceptable approximation for prediction of additive genetic effects. PMID- 14753349 TI - The effect of divergent selection for uterine capacity on prenatal survival in rabbits: maternal and embryonic genetic effects. AB - The aim of this work was to determine whether prenatal survival depends on the genotype of the mother or of the embryo and to identify the critical periods for prenatal mortality in two lines of rabbits divergently selected by high (H) and low (L) uterine capacity. Does from H (n = 124) and L (n = 115) lines were slaughtered at 72 h of gestation. Embryos recovered at 72 h of gestation were transferred to the oviducts of recipient does from the H (n = 23) and L (n = 19) lines. Each recipient does received eight embryos from the H line into one oviduct and eight embryos from the L line into the other. Recipient does were slaughtered on d 28 of gestation. No differences were found between lines in the embryo recovery either in ovulation rate (OR) or in fertilization rate of ova recovered. Recovery rate was higher for the H line (0.80 vs. 0.72, P < 0.01). The number of embryos recovered, fitting ovulation rate as a covariate, was also higher for the H line (9.74 vs. 8.78, P < 0.05). The H line showed a more advanced embryonic stage of development, having a higher percentage of blastocysts (PB) and a lower percentage of compact morulae (PCM) (38% vs. 20%, P < 0.001 for PB, and 51% vs. 64%, P < 0.01 for PCM). The percentage of early morulae was low and similar in both lines. Neither donor nor recipient lines affected embryonic survival from 72 h to 7 d of gestation. Fetal survival was affected by the recipient line (P < 0.05). An interaction between donor and recipient was found. Embryos from the H donor line had a better fetal survival rate than embryos from the L donor line (P < 0.05) in H recipient females. Within L recipient females, embryos from H and L donor lines showed similar fetal survival. Fetal survival was divided into early (from d 7 to 17 of gestation) and late (from d 17 to 28 of gestation). The high recipient line showed a higher early fetal survival than the L recipient line (P < 0.05). The same effect was observed for late fetal survival, but the difference between H and L recipient lines was lower (P < 0.10). Thus, fetal survival depends mainly on the maternal genotype, and the embryo genotype only affects fetal survival when embryo transfer is performed to a favorable maternal environment. Selection for uterine capacity in rabbits leads to modification of early embryonic survival and of early and late fetal survival, but differences are higher for early than for late fetal survival. PMID- 14753350 TI - Comparisons of Angus, Charolais, Galloway, Hereford, Longhorn, Nellore, Piedmontese, Salers, and Shorthorn breeds for weight, weight adjusted for condition score, height, and condition score of cows. AB - Breed differences for weight (CW), height (CH), and condition score (CS) were estimated from records (n = 12,188) of 2- to 6-yr-old cows (n = 744) from Cycle IV of the U.S. Meat Animal Research Center's Germplasm Evaluation (GPE) Program. Cows were produced from mating Angus and Hereford dams to Angus, Hereford, Charolais, Shorthorn, Galloway, Longhorn, Nellore, Piedmontese, and Salers sires. Samples of Angus and Hereford sires were 1) reference sires born from 1962 through 1970 and 2) 1980s sires born in 1980 through 1987. The mixed model included cow age, season of measurement and their interactions, year of birth, pregnancy-lactation code (PL), and breedgroup as fixed effects for CW and CS. Analyses of weight adjusted for condition score included CS as a linear covariate. The model for CH excluded PL. Random effects were additive genetic and permanent environmental effects associated with the cow. Differences among breed groups were significant (P < 0.05) for all traits and were maintained through maturity with few interchanges in ranking. The order of F1 cows for weight was as follows: Charolais (506 to 635 kg for different ages), Shorthorn and Salers, reciprocal Hereford-Angus (HA) with 1980s sires, Nellore, HA with reference sires, Galloway, Piedmontese, and Longhorn (412 to 525 kg for different ages). Order for height was as follows: Nellore (136 to 140 cm), Charolais, Shorthorn, Salers, HA with 1980s sires, Piedmontese, Longhorn, Galloway and HA with reference sires (126 to 128 cm). Hereford and Angus cows with reference sires were generally lighter than those with 1980s sires. In general, breed differences for height followed those for weight except that F1 Nellore cows were tallest, which may in part be due to Bos taurus-Bos indicus heterosis for size. PMID- 14753351 TI - Prevalence of spondylosis deformans and estimates of genetic parameters for the degree of osteophytes development in Italian Boxer dogs. AB - The aim of this study was to assess the prevalence of spondylosis deformans and to investigate genetic aspects of the degree of osteophytes development (DOD) in the Italian Boxer dog population. A total of 849 Boxer dogs was radiographed on the thoracic, lumbar, and sacral regions of the spine and scored for DOD. Grading of DOD was performed for all 20 intervertebral sites comprised within the first thoracic site (site T1-T2) and the site between the seventh lumbar and the first sacral vertebra (site L7-S1). Scores for DOD ranged from 0 (no osteophytes development) to 3 (presence of a bony spur formed by osteophytes on adjoining vertebrae). The first five thoracic sites exhibited no variation for DOD and were not considered in the analysis. The prevalence of spondylosis deformans was 84%, and frequency of dogs showing at least one intervertebral site that scored 3 for DOD was 50%. Scores for DOD at different sites were analyzed as different traits. Nongenetic effects influencing DOD scores were sex, age at screening, and the kennel. Posterior densities of heritability (h2) were estimated using a univariate Bayesian analysis. Eight sites exhibited a posterior probability greater than 0.8 for h2 > 10% and were considered in a multivariate restricted maximum likelihood analysis. Estimated h2 from multivariate analysis ranged from 25 to 48% (SE from 5 to 7%). Three sites exhibited h2 estimates greater than 40%. Genetic correlations for DOD scored at different sites ranged from 0.07 to 0.96. All thoracic sites had estimated correlations larger than 0.85 with other thoracic sites. Genetic correlation between the first and the second lumbar site was 0.91. Correlations between thoracic sites and the first two lumbar sites ranged from 0.5 to 0.9. Sites L6-L7 and L7-S1 also exhibited weak relationships with all remaining sites. Breeding values of dogs for DOD at the eight sites were predicted using estimated covariance matrices. A selection index for DOD was computed from predicted breeding values and a set of relative weighting factors produced by a panel of veterinarians. The index was the most important effect influencing phenotypic differences between dogs for average DOD score, number of affected sites, and number of sites with a DOD score > 1 (P < 0.001). The degree of osteophytes development is a trait showing exploitable additive genetic variance, and breeding programs for decreasing prevalence and severity of spondylosis deformans might focus on this trait. PMID- 14753352 TI - Long-term, but not short-term, treatment with somatotropin during pregnancy in underfed pigs increases the body size of progeny at birth. AB - Treatment of pigs with porcine ST (pST) in early to mid-pregnancy increases body weight and length of their fetuses by mid-pregnancy, but this increased weight may not persist to birth. We investigated the effects of short- (25 d) and long term (75 d) treatment with pST, and interactions between long-term pST treatment and crude protein content of diet, in restricted-fed gilts. In both experiments, Large White x Landrace gilts were bred at first estrus to Large White x Duroc boars and allowed to farrow naturally. In the first experiment, gilts were fed 1.8 kg/d of a diet containing 13.5 MJ DE/kg of DM and 15.05% CP (as-fed basis) throughout pregnancy, and were injected daily with 0, 2, or 4 mg pST from d 25 to 50 of pregnancy. Maternal treatment with pST from d 25 to 50 of pregnancy did not affect the number of piglets born per litter or progeny size at birth. In the second experiment, gilts were injected daily with 0 or 2 mg of pST and fed 2.2 kg/d of a diet containing 14.5 MJ DE/kg and either (as-fed basis) 16.6% (0.81% lysine) or 22.2% CP (1.16% lysine) from d 25 to 100 of pregnancy. All gilts were then fed 3.0 kg/d of the lower protein diet from d 100 of pregnancy to farrowing. Treatment with 2 mg pST/d from d 25 to 100 of pregnancy increased live weight of all gilts during the treatment period (P = 0.016), but the change in maternal live weight from d 25 to 100 of pregnancy was only increased (P = 0.001) by pST in gilts fed the higher protein diet. Live weight of gilts 1 d after farrowing was increased by pST treatment (P = 0.007), but was not altered by protein content of diet during pregnancy. In gilts fed the lower protein diet, but not in those fed the higher protein diet, pST treatment decreased maternal backfat depth during treatment (P < 0.020) and 1 d after farrowing (P = 0.002). Treatment with pST during pregnancy did not affect the number of piglets born per litter but independently increased body weight by 11.6% (P < 0.001) and length by 3.4% (P = 0.005) of progeny at birth and decreased (P < 0.01) the negative effect of litter size on body weight at birth. We conclude that in feed-restricted gilts, fetal weight gains in response to 25 d of pST treatment before mid-pregnancy are not maintained to term but that treatment with pST during most of pregnancy increases progeny size at birth and reduces maternal constraint of fetal growth. PMID- 14753353 TI - Fertilization and blastocyst development in oocytes obtained from prepubertal and adult pigs. AB - Polyspermic fertilization and embryo quality are important issues for the in vitro production of pig embryos. We hypothesized that oocyte donor (prepubertal gilt vs. sow) affects polyspermy and blastocyst development in vitro and that the sexual maturity of the oocyte donor affects the response to sperm concentration in the fertilization medium. In Exp. 1, oocytes of sows and gilts were mounted and stained 12 h after insemination to provide fertilization data. In Exp. 2, putative embryos were developed in vitro to 144 h post-insemination before mounting. In both experiments, cumulus-oocyte complexes (COC) were collected from ovaries of prepubertal gilts and adult sows. Sperm were added after maturation of COC for 40 to 44 h. Sperm from two boars at 0.5 to 5.0 x 10(6) sperm/mL was used for insemination. More (P < 0.01) monospermic fertilizations were observed in oocytes derived from gilts than for oocytes from sows. There were fewer (P < 0.02) penetrated sperm per fertilized oocyte in oocytes from gilts compared with sows. There were effects of semen donor (boar) on the percentage of monospermic (P < 0.01) and polyspermic (P < 0.002) fertilizations, and on the number of penetrated sperm/fertilized oocyte (P < 0.02). In Exp. 2, cleavage and blastocyst formation was evaluated at 2 and 6 d postinsemination, respectively. More (P < 0.001) blastocysts developed from sow-derived oocytes than from gilt-derived oocytes. More (P < 0.05) total cells per blastocyst were observed in embryos from sow-derived oocytes than from gilt-derived oocytes. Semen donor affected the percentage of oocytes cleaving (P < 0.02), and a boar x sperm concentration interaction affected (P < 0.05) the incidence of blastocyt formation. Results indicate that sexual maturity of the donor is not responsible for the high incidence of polyspermy in porcine in vitro fertilization. However, blastocyst development is improved by the use of oocytes from sows rather than from prepubertal gilts. PMID- 14753354 TI - Effects of energy intake and body weight on physical and chemical body composition in growing entire male pigs. AB - Two experiments were conducted to determine independent effects of BW and DE intake on body composition and the partitioning of retained body energy between lipid and protein in pigs with high lean tissue growth potentials and when energy intake limited whole-body protein deposition. In a preliminary N-balance experiment involving 20 entire male pigs at either 30 or 100 kg BW, it was established that whole-body protein deposition increased linearly (P < 0.05) with DE intake at both BW. These results indicate that DE intake controlled whole-body protein deposition and that these pigs did not achieve their maximum whole-body protein deposition when fed semi-ad libitum. In the main serial slaughter experiment, 56 pigs, with a BW of 15 kg, were assigned to one of four DE intake schemes and slaughtered at 40, 65, 90, or 115 kg BW. Within DE intake schemes, DE intake was increased linearly (P < 0.05) with BW, allowing for an assessment of effects of DE intake and slaughter BW on chemical and physical body composition (carcass, viscera, blood). Between 15 and 90 kg BW, average DE intake of 16.1, 20.9, 25.2, and 28.8 MJ/d supported average BW gains of 502, 731, 899, and 951 g/d, respectively. The proportion of whole-body protein present in the carcass increased with BW and decreased with DE intake (P < 0.05), whereas the distribution of whole-body lipid between carcass and viscera was not influenced by BW and DE intake. A mathematical relationship was developed to determine the relationship between DE intake at slaughter (MJ/d) and chemical body composition in these pigs: whole-body lipid-to-protein ratio = 1.236 - 0.056 x (DE intake) + 0.0013 x (DE intake)2, r2 = 0.71. The data suggests that absolute DE intake alone was an adequate predictor of chemical body composition in this population of entire male pigs over the BW and DE intake ranges that were evaluated, simplifying the characterization of this aspect of nutrition partitioning for growth in different pig populations. PMID- 14753355 TI - Growth performance of nursery pigs fed diets containing increasing levels of corn distiller's dried grains with solubles originating from a modern Midwestern ethanol plant. AB - Two experiments were conducted to determine the effects of including distiller's dried grains with solubles in nursery diets on growth performance, and to establish maximum inclusion rates for corn distiller's dried grains with solubles originating from modern, "new-generation" ethanol plants (built since 1990). Ninety-six crossbred pigs (BW = 6.18 +/- 0.14 kg) were blocked by gender and ancestry, and pigs within each block were randomly assigned to one of six dietary treatments (four pigs/pen, four pens/dietary treatment) in each of two growth performance experiments. Dietary treatments provided 0, 5, 10, 15, 20, or 25% distiller's dried grains with solubles during Phases 2 and 3 of a three-phase nursery feeding program for early-weaned pigs. Pigs in Exp. 1 were slightly older (19.0 vs. 16.9 d of age) and heavier (7.10 vs. 5.26 kg) at the beginning of the experiment compared to pigs in Exp. 2. All pigs in both experiments were provided a commercial pelleted Phase 1 diet for the first 4 d after weaning and were then switched to their respective experimental Phase 2 diets, which were fed for 14 d, followed by their respective Phase 3 experimental diets, which were fed for a subsequent 21-d feeding period. Experimental diets were formulated to contain equivalent apparent ileal digestible lysine (1.35 and 1.15%), apparent ileal digestible methionine + cystine (0.80 and 0.65%), ME (3,340 and 3,390 kcal/kg), calcium (0.95 and 0.80%), and total phosphorus (0.80 and 0.70%) within Phases 2 and 3, respectively. Overall growth rate, ending body weight, and feed conversion were similar among pigs regardless of dietary distiller's dried grains with solubles level for both experiments. In Exp. 1, feed intake was not affected by dietary treatment (P > 0.10). In Exp. 2, however, increasing the level of distiller's dried grains with solubles linearly decreased feed intake (P < 0.02) during Phase 2 and tended to decrease voluntary feed intake (P < 0.09) over the length of the experiment. These results suggest that the corn distiller's dried grains with solubles used in this study can be included in Phase 3 diets for nursery pigs at dietary levels of up to 25% without negatively affecting growth performance after a 2-wk acclimation period. Including high levels of corn distiller's dried grains with solubles in diets for pigs weighing less than 7 kg in BW, however, may negatively influence feed intake and growth. PMID- 14753356 TI - Energy requirements of cattle for standing and for ingestion, estimated by a ruminal emptying technique. AB - Energy requirements for ingestion and standing were determined in open-circuit respiration chambers with four ruminally cannulated German Red Pied steers weighing 617 +/- 53 kg of BW (mean +/- SD). The requirement for standing over lying was derived by regressing heat production on time spent standing within 2-h periods when no feed was offered to avoid any interference with eating activity, and amounted to 14 kJ/(d x kg of BW). The energy requirement for ingestion was determined by calculating the difference between heat production during a 2-h period without feed and during a consecutive 2-h period in which straw of different particle sizes, fresh and conserved grass, or rolled barley were offered for ad libitum intake. Before measurements, the rumens of the steers were emptied, washed, and filled with a buffer solution to avoid heat production by metabolism of absorbed nutrients from the feed ingested during the experimental periods. The mean value for all feeds tested was 20 J/(min of ingestion x kg BW). Relating heat production to the amount of DM or fiber ingested did not decrease variation among feeds. This confirms the observations of earlier studies, that energy requirement for ingestion is mainly determined by time spent eating. Results of additional measurements, in which the same amounts of the respective feeds ingested in preceding periods were put into the emptied rumens via the cannulas, showed that the presence of the feed in the rumen did not increase heat production, indicating that the increment of heat production during eating is mainly caused by ingesting and chewing the feed. Further measurements in the same animals with an ingesta-filled rumen showed that ingestion of straw led to an increase in heat production per minute of chewing similar to those with emptied rumens, which confirms the validity of the experimental procedure using ruminally emptied animals to determine the energy requirement for ingestion. PMID- 14753357 TI - Effect of source and amount of energy and rate of growth in the growing phase on adipocyte cellularity and lipogenic enzyme activity in the intramuscular and subcutaneous fat depots of Holstein steers. AB - Seventy-three Holstein steers (initial BW 138.5 +/- 4.3 kg; approximately 3 mo of age) were allotted by BW to one of three growing-phase treatments to determine the effect of source and amount of energy on feedlot performance, and characteristics of subcutaneous (s.c.) and intramuscular (i.m.) adipose tissue. Treatment diets were 1) high concentrate fed ad libitum (ALC); 2) high forage fed ad libitum for 55 d, then a mid-level forage diet fed ad libitum for 98 d (ALF); or 3) limit-fed high concentrate to achieve a gain of 0.8 kg/d for 55 d, then to achieve a gain of 1.2 kg/d for 98 d (LFC). All steers were fed the ALC diet from d 154 to slaughter. Eight steers per treatment were selected after an average of 145 and 334 d on feed for determination of adipocyte cellularity and lipogenic enzyme activity at the end of the growing and finishing phases, respectively. Remaining steers were slaughtered after an average of 334 d on feed. At initial slaughter, ALC steers had a two- to threefold greater (P < 0.05) s.c. fat depth, and 1.9-fold greater (P < 0.01) longissimus muscle ether extract than steers in other groups. At final slaughter, LFC steers had a greater fat depth than ALF steers (P < 0.10) and had the greatest (P < 0.10) longissimus muscle ether extract. Increased fat depth for ALC steers at initial slaughter was a result of a greater (P < 0.05) mean adipocyte diameter in the s.c. depot. Mean i.m. adipocyte diameter followed the same trend (P < 0.16). The number of adipocytes per gram of s.c. fat was least for ALC and greatest for ALF (P < 0.10) at initial slaughter. Mean diameter and number of adipocytes per gram of i.m. and s.c. fat did not differ among treatments at final slaughter (after 180 d on a common finishing diet). High energy (ALC) increased activities of ATP-citrate lyase, fatty acid synthase, 6-phosphogluconate dehydrogenase, glucose-6-phosphate dehydrogenase, and malate dehydrogenase (P < 0.05), in the s.c. depot, and increased activities of ATP-citrate lyase and glucose-6-phosphate dehydrogenase (P < 0.10) in the i.m. depot at initial slaughter. Lipogenic enzyme activity in the s.c. depot at final slaughter did not differ among treatments. Glucose-6 phosphate dehydrogenase activity in the i.m. depot at final slaughter was lowest (P < 0.10) in ALF. Hypertrophy made a greater contribution to fat tissue growth than hyperplasia. Hypertrophy was affected by amount of energy, whereas hyperplasia was affected by source of energy. Differences diminished when cattle were fed the common finishing diet. PMID- 14753358 TI - A mechanistic model for predicting the nutrient requirements and feed biological values for sheep. AB - The Cornell Net Carbohydrate and Protein System (CNCPS), a mechanistic model that predicts nutrient requirements and biological values of feeds for cattle, was modified for use with sheep. Published equations were added for predicting the energy and protein requirements of sheep, with a special emphasis on dairy sheep, whose specific needs are not considered by most sheep-feeding systems. The CNCPS for cattle equations that are used to predict the supply of nutrients from each feed were modified to include new solid and liquid ruminal passage rates for sheep, and revised equations were inserted to predict metabolic fecal N. Equations were added to predict fluxes in body energy and protein reserves from BW and condition score. When evaluated with data from seven published studies (19 treatments), for which the CNCPS for sheep predicted positive ruminal N balance, the CNCPS for sheep predicted OM digestibility, which is used to predict feed ME values, with no mean bias (1.1 g/100 g of OM; P > 0.10) and a low root mean squared prediction error (RMSPE; 3.6 g/100 g of OM). Crude protein digestibility, which is used to predict N excretion, was evaluated with eight published studies (23 treatments). The model predicted CP digestibility with no mean bias (-1.9 g/100 g of CP; P > 0.10) but with a large RMSPE (7.2 g/100 g of CP). Evaluation with a data set of published studies in which the CNCPS for sheep predicted negative ruminal N balance indicated that the model tended to underpredict OM digestibility (mean bias of -3.3 g/100 g of OM, P > 0.10; RMSPE = 6.5 g/100 g of OM; n = 12) and to overpredict CP digestibility (mean bias of 2.7 g/100 g of CP, P > 0.10; RMSPE = 12.8 g/100 g of CP; n = 7). The ability of the CNCPS for sheep to predict gains and losses in shrunk BW was evaluated using data from six studies with adult sheep (13 treatments with lactating ewes and 16 with dry ewes). It accurately predicted variations in shrunk BW when diets had positive N balance (mean bias of 5.8 g/d; P > 0.10; RMSPE of 30.0 g/d; n = 15), whereas it markedly overpredicted the variations in shrunk BW when ruminal balance was negative (mean bias of 53.4 g/d, P < 0.05; RMSPE = 84.1 g/d; n = 14). These evaluations indicated that the Cornell Net Carbohydrate and Protein System for Sheep can be used to predict energy and protein requirements, feed biological values, and BW gains and losses in adult sheep. PMID- 14753359 TI - Fermentation of eastern gamagrass (Tripsacum dactyloides [L.] L.) by mixed cultures of ruminal microorganisms with or without supplemental corn. AB - Five dual-flow fermentors (700 mL) were used to determine the effects of eastern gamagrass (Tripsacum dactyloides [L.] L.) diets on microbial metabolism by mixed rumen cultures. Fermentors were incubated with filtered ruminal contents and allowed to adapt for 4 d to diets followed by 3 d of sample collection. Five dietary treatments were tested: 1) gamagrass hay (GH) + no corn (GHNC), 2) gama grass silage (GS) + no corn (GSNC), 3) GS + low corn (GSLC), 4) GS + medium corn (GSMC); and 5) GS + high corn (GSHC). The experiment was conducted as a randomized complete block design with five treatments and three replications. Total VFA concentrations were not affected by diets. Corn addition linearly decreased (P < 0.001) molar proportion of acetate. In contrast, molar proportion of propionate was reduced in GSLC (cubic effect, P < 0.001) but remained similar across other diets. Corn supplementation linearly increased molar proportion of butyrate (P < 0.001). The acetate + butyrate-to-propionate ratio was highest in cultures offered GSLC (cubic effect, P < 0.001) but similar across other diets. Feeding GSNC resulted in a higher ruminal pH compared with GHNC (P < 0.03). Increasing the level of corn supplementation in GS linearly decreased culture pH (P < 0.001). All diets resulted in similar methane production, with the exception of GSMC, which lowered methane output (quadratic effect, P < 0.004). Total substrate fermented to VFA and gas tended to be greater with GHNC than with GSNC (P < 0.06) and linearly increased with the addition of corn (P < 0.004). Neutral detergent fiber digestibility was similar between GH and GS and was not affected by supplemental corn. Microbial N flow increased in cultures offered GSHC (quadratic effect, P < 0.02). Corn supplementation at the medium and high level linearly decreased C 18:0 (P < 0.02) and increased trans-C18:1 (P < 0.004). Including corn at the high level with GS did not have a detrimental effect on fermentation in dual-flow fermentors. PMID- 14753360 TI - Technical note: a procedure for the preparation and quantitative analysis of samples for titanium dioxide. AB - A procedure was developed for the rapid analysis of titanium dioxide (TiO2) concentrations in feed and fecal samples. Samples were digested in concentrated H2SO4 for 2 h, followed by addition of 30% H2O2, and absorbance was measured at 410 nm. Standards were prepared by spiking blanks with increasing amounts of TiO2, resulting in a linear standard curve. Complete analysis using this procedure can typically be accomplished within 4.5 h. This procedure was compared to a previously published dry-ash procedure for the analysis of TiO2 in bovine fecal samples. Three sources of OM devoid of TiO2 (a forage sample, a bovine fecal sample without Cr2O3, and a bovine fecal sample containing Cr2O3) were spiked with graded amounts (0, 2, 4, 6, 8, or 10 mg) of TiO2. With our procedure, TiO2 recoveries averaged 96.7, 97.5, and 98.5%, for the three OM sources, respectively, vs. 74.3, 83.8, and 53.1% for the same samples analyzed using the dry-ash method. These results suggest that our procedure is a rapid and accurate alternative to dry-ash procedures for the determination of TiO2. PMID- 14753361 TI - Effect of live weight gain of steers during winter grazing: II. Visceral organ mass, cellularity, and oxygen consumption. AB - Two experiments were conducted to examine the effect of BW gain during winter grazing on mass, cellularity, and oxygen consumption of splanchnic tissues before and after the feedlot finishing phase. In each experiment, 48 fall-weaned Angus x Angus-Hereford steer calves were assigned randomly to one of three treatments: 1) high rate of BW gain grazing winter wheat (HGW), 2) low rate of BW gain grazing winter wheat (LGW), or 3) grazing dormant tallgrass native range supplemented with 0.91 kg/d of a 41% CP supplement (NR). At the end of winter grazing, four steers were selected randomly from each treatment for initial slaughter to measure organ mass, cellularity, and oxygen consumption. All remaining steers were placed into a feedlot and fed to the same backfat end point (1.27 cm). Six steers were selected randomly from each treatment for final organ mass, cellularity, and oxygen consumption. Initial empty BW (EBW) was greatest (P < 0.001) for HGW, intermediate for LGW, and least for NR steers in both Exp. 1 and 2 (355 > 263 > 207 +/- 6.5 kg and 337 > 274 > 205 +/- 8.7 kg, respectively). For both experiments, the initial total gastrointestinal tract (GIT; g/kg of EBW) proportional weight was greater (P < 0.05) in NR steers than in LGW, and LGW steers had greater (P < 0.05) initial GIT proportional weight than HGW steers. Proportional weight of total splanchnic tissues (TST; g/kg of EBW) did not differ (P < 0.19) among treatments. Initial duodenal RNA concentration and RNA:protein were greater (P < 0.02) in LGW than in HGW steers, and NR steers were intermediate. Initial in vitro liver O2 consumption was greater (P < 0.09) in HGW and LGW than in NR steers (34.5 > 16.9 mL/min), whereas initial small intestinal oxygen consumption was greater (P < 0.01) in LGW than in HGW and NR steers (12.1 > 5.2 mL/min). Ruminal papillae oxygen consumption did not differ (P < 0.55) among treatments. The rate of decrease of GIT (g x g EBW(-1) x d(-1)) during finishing was greater in NR than in HGW and LGW steers in both Exp. 1 and 2, but mesenteric fat (g x g EBW(-1) x d(-1)) increased for NR steers, resulting in a similar (P < 0.75) increase in TST across the finishing period for all treatments. Similar rates of increase in TST across the finishing phase corresponded with similar rates of live and carcass weight gain among treatments. Our data support the hypothesis that increased visceral organ mass increases maintenance energy requirements of growing cattle. PMID- 14753362 TI - Effect of alkali pretreatment of wheat straw on the efficacy of exogenous fibrolytic enzymes. AB - The effects of pretreating wheat straw with alkali on the efficacy of exogenous fibrolytic enzymes for improving straw digestibility were studied in vitro, in situ, and in vivo. In Exp. 1, untreated straw (US); alkali-treated (5% NaOH, wt/wt) straw (AS); and autoclaved, alkali-treated straw (AAS) were sprayed with 0 or 1.5 mg/g DM of enzyme mix (xylanase, beta-glucanase, carboxymethylcellulase, and amylase) and incubated for 30 h in buffered ruminal fluid (3 x 2 factorial arrangement). Enzymes increased (P < 0.001) gas production and the incorporation of 15N into microbial N at 4 h, more so with AS or AAS than with US (P < 0.001 for gas; P < 0.05 for 15N). In Exp. 2, US and AS were sprayed with enzymes at 0, 0.15, or 1.5 mg/g DM (2 x 3 factorial) and incubated ruminally in nylon bags for up to 80 h to determine the in situ DM disappearance (ISDMD). Interactive effects (P < 0.05) of pretreatment and enzymes were observed on all ruminal degradation parameters. Alkali increased the rate (P < 0.01) and extent (P < 0.001) of ISDMD irrespective of enzymes. Enzymes applied to US did not affect the extent of ISDMD, but they increased (P < 0.01) the extent of ISDMD when applied to AS. Substrates from Exp. 1 and 2 were incubated in acetate buffer for 24 h to measure the hydrolytic loss of DM and release of reducing sugars and phenolic compounds. Alkali pretreatment and enzymes each increased (P < 0.001) DM loss and the release of reducing sugars and, in combination, exerted synergistic effects (P < 0.001). Enzymes did not affect the release of phenolic compounds from the straw. In Exp. 3, total-tract digestibility of untreated and enzyme-treated (100 mL/kg DM) ammoniated straw was assessed using 32 beef cows in eight pens. Wrapped straw bales were injected with NH3 (3% [wt/wt], DM basis) 4 mo before the study; enzymes were applied immediately before feeding. Applying enzyme to ammoniated straw increased (P < 0.05) digestibilities of DM, OM, and total N but did not affect the intake of DM or digestibility of ADF. Pretreatment of straw with alkali enhanced the efficacy of exogenous enzymes, presumably by breaking esterified bonds and releasing phenolic compounds and/or by swelling the crystalline cellulose and enhancing enzyme penetration. Including enzymes that mimic alkali hydrolysis (e.g., esterases) in commercial feed additives could substantially improve the value of these products for ruminants. PMID- 14753363 TI - Environmental effects on pig performance, meat quality, and muscle characteristics. AB - The objective of this experiment was to determine the effect of diverse production systems on pig performance, muscle characteristics, and their relation to pork quality measures. Birth and rearing conditions were evaluated using 48 barrows during the fall/winter months. Pigs were farrowed in either indoor crates or outdoor huts. At weaning, indoor- and outdoor-born pigs were allotted randomly to treatments arranged in a 2 x 2 factorial design with two birth (indoor vs. outdoor) and rearing (indoor vs. outdoor) environments. Pigs housed indoors were on concrete-slatted flooring (1.2 m2/pig), and pigs housed outdoors were on an alfalfa pasture (212 m2/pig). Body weight data were collected. Muscle samples were removed within 1 h postmortem from the longissimus (LM) and semimembranosus (SM) muscles. Muscle samples were stained histochemically to identify type I, IIA, and IIB/X muscle fibers. Boneless loins were collected from the left side of each carcass and aged for 14 d. Objective and subjective color measurements were taken on the longissimus muscle at the 10th rib on d 14 postmortem. Loin chops were evaluated for sensory attributes, shear force, and retail display features. Pigs born outdoors were heavier and had a greater ADG at most growth intervals postweaning (d 28, 56, and 112; P < 0.05) than pigs born indoors. Pigs reared outdoors were heavier (P = 0.02) at d 140 (120.1 vs. 112.9 +/- 4.9 kg), and had higher (P = 0.01) ADG (2.2 vs. 1.9 kg/d) and gain:feed ratios (0.41 vs. 0.37) than did pigs reared indoors. Birth x rearing environment interactions were not significant (P > 0.10) for most measures. Carcass and meat quality measures did not differ (P > 0.05) among treatment groups, but loin chops from outdoor born or reared pigs had higher (P < 0.05) a* values than chops from pigs born or reared indoors. The LM muscle of pigs born outdoors had a higher (P < 0.01) percentage of type I, and a lower (P < 0.05) percentage of type IIA fibers than did pigs born indoors. Pigs reared outdoors had a higher (P < 0.01) percentage of IIA fibers and a lower (P < 0.05) percentage of IIB/X fibers than did pigs reared indoors for the LM and SM muscles. Outdoor production systems may influence growth, pork color, and muscle fiber types. PMID- 14753364 TI - Improving pork quality by feeding supranutritional concentrations of vitamin D3. AB - Duroc-cross pigs (n = 25) were assigned to one of three experimental finishing diets containing 0 (control), 40,000 (40), or 80,000 (80) IU of supplemental vitamin D3/kg of feed (as-fed basis)to test the effects of vitamin D3 on pork quality traits. Experimental diets were fed for 44 or 51 d before slaughter, and days on feed were blocked in the experimental design. A trend existed for pigs receiving the highest concentration of vitamin D3 supplementation to have a lower (P = 0.08) ADG (0.77 kg/d) compared with pigs fed either the 40-diet (0.88 kg/d) or control (0.92 kg/d). Diet did not (P > 0.10) affect backfat thickness measured along the midline, 10th-rib fat depth, longissimus muscle area, muscle score, or hot carcass weights. Longissimus pH, measured at 0.5, 1, 2, 3, 4, and 24 h postmortem, was higher (P < 0.05) for pigs on the 80-diet than those fed the control diet. Longissimus muscle color, measured at 24 h postmortem, from pigs fed either the 40- or 80-diet were darker (lower L* values; P < 0.05) than those fed the control diet. Objective longissimus color scores were higher (P < 0.01), and firmness/wetness scores lower (P < 0.05), for pigs on the 80-diet as compared to those on the 40-diet or control diet. The diet had no (P > 0.10) effect on Warner-Bratzler shear force values; percentage of cook loss; or trained sensory panel evaluations for tenderness, juiciness, and flavor. Feeding the 80-diet increased (P < 0.05) plasma vitamin D3 and calcium after 2, 4, and 6 wk on feed compared with the control diet. Vitamin D3 and 25-hydroxy vitamin D3 concentrations in the longissimus muscle increased (P = 0.001) with increasing vitamin D3 levels in the diet; however, muscle calcium concentrations and fiber type were not (P > 0.30) affected by diet. These results indicate that feeding supranutritional levels of vitamin D3 for at least 44 d improves pork color and increases pH, but may retard growth if fed at 80,000 IU/kg of feed. PMID- 14753365 TI - Vitamin D3 supplementation of cull cows: effects on longissimus and semitendinosus muscle tenderness. AB - Previous studies have shown that supplementation of vitamin D3 to cow diets for 4 to 10 d before slaughter lowers Warner-Bratzler shear force (WBSF) values and increases sensory tenderness scores in beef cuts. The present study was conducted to evaluate the effects of vitamin D3 supplementation on muscle calcium concentration, WBSF values, and sensory tenderness ratings of LM and semitendinosus (ST) muscles from cull, predominately Angus, cows (eight cows per treatment). Treatments included 0 (control), 5 million IU, or 7.5 million IU of vitamin D3 supplemented daily for 7 d preslaughter. Twenty-four hours after slaughter, 2.54-cm-thick LM and ST muscle steaks were cut; aged for either 0, 7, 14, or 21 d (ST steaks aged for 7 d only); and frozen at -20 degrees C for WBFS and sensory analysis. Mean values for LM calcium concentration tended to increase (P = 0.14) with vitamin D3 supplementation (154, 176, and 183 microig/g, fresh basis, for 0, 5, and 7.5 million IU/d, respectively). After 7 d of aging, LM steaks from cows fed 7.5 million IU had lower (P < 0.05) WBSF values than 7-d steaks from controls and cows fed 5.0 million IU/d aged 7 d; however, vitamin D3 supplementation had no (P > 0.05) effect on WBSF values of ST steaks aged 7 d. Vitamin D3 supplementation did not (P > 0.05) affect sensory tenderness ratings for either LM or ST steaks at any aging period. Aging, however, had a linear (P < 0.001) effect on tenderness, with an increase in tenderness as aging time increased from 0 to 21 d. Thus, results from the present study indicate that vitamin D3 supplementation, at these levels and duration before slaughter, provided little benefit to muscle tenderness of beef from cull cows. PMID- 14753366 TI - Insulin and dexamethasone regulate stearoyl-CoA desaturase mRNA levels and fatty acid synthesis in ovine adipose tissue explants. AB - Sheep adipose tissue explants were maintained in culture for 24 h in the presence of insulin, dexamethasone, or insulin and dexamethasone, and stearoyl-CoA desaturase (SCD) messenger RNA (mRNA) levels and fatty acid synthesis were measured. Insulin increased SCD mRNA levels (P = 0.008) and synthesis of both saturated (P = 0.07) and unsaturated (P < 0.001) fatty acids but had the greatest effect on unsaturated fatty acid synthesis, resulting in the overall production of a greater (P < 0.001) proportion of monounsaturated fat. Dexamethasone, alone, had the opposite effect but actually potentiated the effect of insulin in stimulating SCD expression and both saturated and monounsaturated fatty acid synthesis, without affecting the relative proportions of each. Across adipose tissue depots, the effect of hormones was similar, although the increase in SCD mRNA levels (P = 0.008) and monounsaturated fatty acid synthesis (P < 0.001) was greater in subcutaneous adipose tissue than in the internal (omental and perirenal) depots. These data clearly show that, in ovine adipose tissue, changes in SCD gene expression in response to insulin and dexamethasone are associated with changes in monounsaturated fatty acid synthesis and suggest that it may be possible to develop strategies to manipulate sheep tissues to produce a less saturated fatty acid profile. PMID- 14753367 TI - Technical note: use of belt grill cookery and slice shear force for assessment of pork longissimus tenderness. AB - The present experiments were conducted to determine whether improved beef longissimus shear force methodology could be used to assess pork longissimus tenderness. Specifically, three experiments were conducted to: 1) determine the effect of belt grill (BG) cookery on repeatability of pork longissimus Warner Bratzler shear force (WBSF), 2) compare the correlation of WBSF and slice shear force (SSF) with trained sensory panel tenderness ratings, and 3) estimate the repeatability of pork longissimus SSF for chops cooked with a BG. In Exp. 1 and 2, the longissimus was removed from the left side of each carcass (Exp. 1, n = 25; Exp. 2, n = 23) at 1 d postmortem and immediately frozen to maximize variation in tenderness. In Exp. 1, chops were cooked with either open-hearth electric broilers (OH) or BG, and WBSF was measured. Percentage of cooking loss was lower (P < 0.001) and less variable for chops cooked with a BG (23.2%; SD = 1.7%) vs. OH (27.6%; SD = 3.0%). Estimates of the repeatability of WBSF were similar for chops cooked with OH (0.61) and BG (0.59). Although significant (P < 0.05), differences in WBSF (4.1 vs. 3.9 kg) between cooking methods accounted for less than 5% of the total variation in WBSF. In Exp. 2, the correlation of SSF (r = -0.72; P < 0.001) with trained sensory panel tenderness ratings was slightly stronger than the correlation of WBSF (r = -0.66; P < 0.001) with trained sensory panel tenderness ratings, indicating that the two methods had a similar ability to predict tenderness ratings. In Exp. 3, duplicate samples from 372 carcasses at 2 and 10 d postmortem were obtained, cooked with BG, and SSF was determined. The repeatability of SSF was 0.90, which is comparable to repeatability estimates for beef and lamb. Use of BG cookery and SSF could facilitate the collection of accurate pork longissimus tenderness data. Time and labor savings associated with BG cookery and the SSF technique should help to decrease research costs. PMID- 14753368 TI - The use of vitamin D3 and its metabolites to improve beef tenderness. AB - Three experiments were conducted to determine whether feeding 25-hydroxyvitamin D3 (25-OH D3) or 1,25-dihydroxyvitamin D3 (1,25-(OH)2 D3) improves the tenderness of longissimus dorsi (LD), semimembranosus (SM), and infraspinatus (IF) muscles similar to supplemental vitamin D3 without leaving residual vitamin D3 and its metabolites in muscle. In the first two experiments, 24 crossbred steers were used to determine the effects of different oral amounts of 1,25-(OH)2 D3 (Exp. 1; n = 12) and 25-OH D3 (Exp. 2; n = 12) on plasma Ca2+ concentrations. In the third experiment, crossbred steers were allotted randomly to one of four treatments: 1) control placebo (n = 7); 2) 5 x 10(6) IU of vitamin D3/d (n = 9) for 9 d and harvested 2 d after last treatment; 3) single, 125-mg dose of 25-OH D3 (n = 8) 4 d before harvest; or 4) single, 500-microg dose of 1,25-(OH)2 D3 (n = 9) 3 d before harvest. The LD and SM steaks from each animal were aged for 8, 14, or 21 d, whereas steaks from the IF were aged for 14 or 21 d. All steaks were analyzed for tenderness by Warner-Bratzler shear force and for troponin-T degradation by Western blot analysis. Supplementing steers with vitamin D3 increased (P < 0.01) the concentration of vitamin D3 and 25-OH D3 in all muscles sampled. Feeding steers 25-OH D3 increased (P < 0.05) the concentration of 25-OH D3 in meat, but to an amount less than half that of cattle treated with vitamin D3. Supplemental 1,25-(OH)2 D3 did not affect (P < 0.10) shear force values; however, there was a trend (P < 0.10) for supplemental vitamin D3 and 25-OH D3 to produce LD steaks with lower shear values after 8 and 14 d of aging, and lower (P < 0.10) shear force values for the SM aged for 21 d. Analysis of Western blots indicated that LD steaks from cattle supplemented with vitamin D3 and 25-OH D3 had greater (P < 0.05) troponin-T degradation. Antemortem supplementation of 25-OH D3 seems to increase postmortem proteolysis and tenderness in the LD and SM without depositing large concentrations of residual vitamin D3 and its metabolite 25-OH D3. PMID- 14753369 TI - Evaluation of methods to reduce bacteria concentrations in spray-dried animal plasma and its effects on nursery pig performance. AB - Four experiments with 1,040 weanling pigs (17 +/- 2 d of age at weaning) were conducted to evaluate the effects of spray-dried animal plasma source, drying technique, and methods of bacterial reduction on nursery pig performance. In Exp. 1, 180 barrows and gilts (initial BW 5.9 +/- 1.8 kg) were used to compare effects of animal plasma, animal plasma source, drying technique (spray-dried or freeze dried), and plasma irradiation in nursery pig diets. From d 0 to 10, pigs fed diets containing irradiated spray-dried animal plasma had increased ADG and ADFI (P < 0.05) compared with pigs fed diets containing nonirradiated spray-dried animal plasma. Pigs fed irradiated animal plasma Sources 1 and 2 were similar in ADG and ADFI, but pigs fed animal plasma Source 1 had greater ADG (P < 0.05) than pigs fed animal plasma Source 2 and pigs not fed plasma. Pigs fed freeze-dried animal plasma had growth performance similar (P > 0.36) to pigs fed spray-dried animal plasma. Overall (d 0 to 24), pigs fed irradiated spray-dried animal plasma were heavier (P < 0.05) than pigs fed no animal plasma, whereas pigs fed nonirradiated spray-dried plasma were intermediate. In Exp. 2, 325 barrows and gilts (initial BW 5.8 +/- 1.7 kg) were used to compare the effects of irradiation or formaldehyde treatment of animal plasma and formaldehyde treatment of the whole diet. Pigs fed diets containing irradiated animal plasma had greater ADG (P < 0.05) than pigs fed nonirradiated plasma. Pigs fed formaldehyde-treated plasma had greater ADG and ADFI (P < 0.05) than pigs fed diets with either nonirradiated plasma or whole diet treated with formaldehyde. In Exp. 3 (360 barrows and gilts; initial BW 6.3 +/- 2.7 kg) and Exp. 4 (175 barrows and gilts; initial BW 6.1 +/- 1.7 kg), the irradiation of feed (high bacteria) and food-grade (low bacteria) animal plasma in nursery pig diets was examined. Pigs fed irradiated feed-grade plasma Product 2 had increased ADG (P < 0.05) compared with pigs fed nonirradiated plasma Product 2 and pigs fed the control diet without plasma. In Exp. 3 and 4, pigs fed irradiated food-grade plasma had growth performance similar to pigs fed nonirradiated food-grade plasma (P > 0.12). These studies indicate that bacterial reduction of feed-grade, but not food-grade animal plasma, improves nursery pig performance. PMID- 14753370 TI - Effect of live weight gain of steers during winter grazing: I. Feedlot performance, carcass characteristics, and body composition of beef steers. AB - Two experiments were conducted to examine the effect of previous BW gain during winter grazing on subsequent growth, carcass characteristics, and change in body composition during the feedlot finishing phase. In each experiment, 48 fall weaned Angus x Angus-Hereford steer calves were assigned randomly to one of three treatments: 1) high rate of BW gain grazing winter wheat (HGW), 2) low rate of BW gain grazing winter wheat (LGW), or 3) grazing dormant tallgrass native range (NR) supplemented with 0.91 kg/d of cottonseed meal. Winter grazing ADG (kg/d) for HGW, LGW, and NR steers were, respectively, 1.31, 0.54, 0.16 (Exp. 1) and 1.10, 0.68, 0.15 (Exp. 2). At the end of winter grazing, four steers were selected randomly from each treatment to measure initial carcass characteristics and chemical composition of carcass, offal, and empty body. All remaining steers were fed a high-concentrate diet to a common backfat end point. Six steers were selected randomly from each treatment for final chemical composition, and carcass characteristics were measured on all steers. Initial fat mass and proportion in carcass, offal, and empty body were greatest (P < 0.001) for HGW, intermediate for LGW, and least for NR steers in both experiments. Live BW ADG and gain efficiency during the finishing phase did not differ (P = 0.24) among treatments, but DMI (% of mean BW) for NR and LGW was greater (P < 0.003) than for HGW steers. Final empty-body composition did not differ (P = 0.25) among treatments in Exp. 1. In Exp. 2, final carcass and empty-body fat proportion (g/kg) was greater (P < 0.03) for LGW and NR than for HGW steers. Accretion of carcass fat free organic matter was greater (P < 0.004) for LGW than for HGW and NR steers in Exp. 1, but did not differ (P = 0.22) among treatments in Exp. 2. Fat accretion in carcass, offal, and empty body did not differ (P = 0.19) among treatments in Exp. 1, but was greater (P < 0.05) for LGW and NR than for HGW steers in Exp. 2. Heat production by NR steers during finishing was greater (P < 0.02) than by HGW steers in Exp. 1 and 2. Differences in ADG during winter grazing and initial body fat content did not affect rate of live BW gain or gain efficiency during finishing. Feeding steers to a common backfat thickness end point mitigated initial differences in carcass and empty-body fat content. However, maintenance energy requirements during finishing were increased for nutritionally restricted steers that were wintered on dormant native range. PMID- 14753371 TI - Effect of source and amount of energy and rate of growth in the growing phase on performance and carcass characteristics of early- and normal-weaned steers. AB - One hundred eighty-four Angus x Simmental steers (initial BW 161.7 +/- 3.4 kg) were used to determine whether different sources and amounts of energy in the growing phase could extend the growth curve and maintain high amounts of intramuscular fat deposition in early-weaned steers. Steers were allotted by source, age, and BW to one of four growing-phase (119 to 259 d of age) regimens. For three regimens, steers were weaned at 119 d of age and either 1) fed (DM basis) a 50% grain diet ad libitum (ALC); 2) limit-fed a 70% grain diet to achieve a gain of 0.8 kg/d from 119 to 192 d of age, and 1.2 kg/d from 193 to 259 d of age (LFC); or 3) fed a 60% haylage diet ad libitum from 119 to 192 d of age, and a 25% haylage diet ad libitum from 193 to 259 d of age (ALF). For the fourth regimen, steers were normal-weaned at 204 d of age and fed a silage diet from 205 to 259 d of age (NW). From 260 d of age to slaughter, all steers consumed a 70% grain (DM basis) diet. Limit-fed and ALF steers spent the most, and NW the least amount of time (P < 0.01) in the feedlot to achieve a target fat depth of 1.27 cm. Ad libitum-fed steers were the youngest (331 d), and NW the oldest (383 d) at slaughter (P < 0.01). Overall ADG was greatest for ALC and least for NW steers (P < 0.01). Overall, ALF steers consumed the most DM (P < 0.01). While in the feedlot, LFC and ALC steers were more efficient in converting feed to BW gain (P < 0.01) than ALF and NW steers. Normal-weaned had the least and ALC the greatest (P < 0.01) fat depth at 260 d of age. Consequently, NW steers produced the heaviest, and ALC the lightest (P < 0.01) carcasses at slaughter. Normal-weaned steers had the largest, and ALC and LFC steers had the smallest longissimus muscle area (P < 0.06). Growing phase dietary treatments did not affect (P > 0.20) yield grade. Marbling score did not differ (P > 0.35), but laboratory analysis revealed that ALC steers had the lowest percentage of fat (P < 0.02) in the longissimus muscle. Shear force was greatest (P < 0.08) for steaks from ALC and LFC steers, and least for steaks from ALF and NW steers. Feeding steers the ALC diet from 119 to 260 d of age hastened physiological maturity, decreased marbling scores, and decreased muscle tenderness compared with forage feeding (ALF, NW). Limit-feeding a high-grain diet also hastened physiological maturity and decreased muscle tenderness but did not decrease marbling scores. Source and amount of energy affected partitioning of fat deposition. PMID- 14753372 TI - Effect of increasing levels of undegradable intake protein on metabolic and endocrine factors in estrous cycling beef heifers. AB - To determine the influence of three levels of undegradable intake protein (UIP) supplementation on metabolic and endocrine factors that influence reproduction, 23 yearling crossbred heifers (body condition score = 4.5 +/- 0.5; initial BW = 362 +/- 12 kg) were stratified by BW and assigned randomly to one of three supplements: 1) low UIP (1,135 g x heifer(-1) x d(-1); 30% CP, 115 g UIP, n = 7); 2) mid UIP (1,135 g x heifer(-1) x d(-1); 38% CP, 216 g UIP, n = 8); or 3) high UIP (1,135 g x heifer(-1) x d(-1); 46% CP, 321 g UIP, n = 8). Heifers were estrually synchronized before initiation of supplementation. Supplement was individually fed daily for 30 to 32 d, at which time heifers were slaughtered (d 12 to 14 of the estrous cycle) and tissues collected. Heifers were fed a basal diet of sudan grass hay (6.0% CP) ad libitum. On d 28 of supplementation (d 10 of the estrous cycle), no differences were observed (P > 0.10) in serum insulin or IGF-I among treatments. At slaughter (d 10 to 12 of the estrous cycle), treatments did not influence corpus luteum weight, cerebral spinal fluid leptin, or IGFBP; serum estradiol-17beta, progesterone, leptin, IGF-I, and IGFBP; or anterior pituitary content of IGFBP (P > 0.10). Follicular fluid IGFBP-2 and IGFBP-4 were greater in high-UIP heifers than low- or mid-UIP heifers on d 12 to 14 of the estrous cycle (P < 0.05). Basal serum LH concentrations and LH area under the curve (every 15 min for 240 min) did not differ (P > 0.10) following 28 d of supplementation (d 10 of the estrous cycle); however, basal serum FSH concentrations were greater (P = 0.06) in low- and mid- vs. high-UIP heifers (5.2 and 5.2 vs. 4.6 ng/mL, respectively), and FSH area under the curve was greater (P = 0.03) in low- vs. high-UIP heifers. At slaughter (d 12 to 14 of the estrous cycle), anterior pituitary LH and FSH content and steady-state mRNA encoding alpha, LHbeta, and GnRH receptor did not differ (P > 0.10) among treatments. However, FSHbeta mRNA was increased approximately twofold (P = 0.03) in mid vs. low UIP. In summary, low and mid levels of UIP supplements fed to estrous cycling beef heifers seemed to enhance pituitary expression and/or secretion of FSH relative to high levels of UIP. Moreover, high-UIP supplementation was associated with increased low-molecular-weight IGFBP compared with supplementation of low and mid levels of UIP. These data suggest that differing levels of UIP supplementation may alter pituitary and ovarian function, thereby influencing reproductive performance in beef heifers. PMID- 14753373 TI - Effects of different grazing and feeding periods on performance and carcass traits of beef steers. AB - Over three consecutive years, 180 (60/yr) fall-born steer calves were weaned in May (average initial BW = 238 kg, SD = 36.2 kg) and allocated to one of three groups: 1) calf-fed steers that entered the feedlot at weaning; 2) short yearlings that grazed irrigated pasture for another 4 mo and entered the feedlot in September; and 3) long yearlings that grazed with short yearlings during the summer, remained on annual California foothills range through the fall, winter, and spring, and entered the feedlot the following May. All steers were fed until the average group backfat (BF), determined by ultrasound, reached 11 to 12 mm. On pasture, short- and long-yearling steers gained weight in the summer; long yearlings then slightly lost weight in the fall and winter, and then gained weight again the following spring. Average days in the feedlot were 188, 158, and 94 (P < 0.10) for calves, short yearlings, and long yearlings, respectively. Feedlot DMI increased with age (and weight) at feedlot entry, with no difference among groups in gain:feed ratio. The gain of BF was nil on pasture, even when animals were gaining weight, and then increased rapidly when animals were placed on a high-energy diet. Final body weights were heaviest (P < 0.10) in long yearlings, followed by short yearlings and then calves, indicating that a prolonged growing period increases the apparent mature size of the animal. Moreover, total carcass fat contents and percentage of Choice or above were all lower (P < 0.10) in cattle that were older at feedlot entry (i.e., long yearlings) compared with the other groups. In conclusion, increasing the backgrounding period decreased time and total concentrate requirements in the feedlot of Angus-Hereford steers. Older cattle reached 10 mm of BF at heavier weights. Grazing animals gained weight without increasing BF; however, BF increased rapidly in the feedlot. Prolonged grazing may decrease quality grade, either by impairing the ability of the animal to deposit intramuscular fat or by decreasing the time during which dietary energy supply is adequate for intramuscular fat deposition to occur. PMID- 14753374 TI - Self-limiting supplements fed to cattle grazing native mixed-grass prairie in the northern Great Plains. AB - Objectives of this research were to compare animal performance with or without supplementation, compare effectiveness of three intake limiters, and to examine seasonal changes in nutritive value of native range in south-central North Dakota. Treatments included 1) control (CONT; no supplement); 2) hand-fed (HF) supplement, with no chemical limiter; 3) 16% salt (NACL); 4) 5.25% ammonium chloride and ammonium sulfate (AS); and 5) 7% calcium hydroxide (CAOH). Supplements were based on wheat middlings, barley malt sprouts, and soybean hulls and were formulated to provide 40% of the CP intake and 32% of the NEm intake of 350-kg steers. Trials 1 and 2 each used 70 yearling steers (370.8 +/- 0.04 and 327.9 +/- 0.76 kg initial BW for Trials 1 and 2, respectively). In each year, four 28-d periods from the latter half of June through mid-October were used. Steers were stratified by weight and allotted randomly to treatments in 1 of 10 16-ha pastures (two pastures per treatment for each trial). In Trial 1, diet sampling began in the first 28-d period, but supplementation did not begin until the second 28-d period. In Trial 2, supplementation and diet collection began in the first 28-d period. Cation-anion differences (DCAD; Na + K - Cl - S) for NACL, AS, CAOH, and HF supplements were 151, -735, 160, and 166 mEq/ kg, respectively. In Trial 1, no treatment, period, or treatment x period effects for supplement intake were detected (P > or = 0.29). In Trial 2, a treatment x period interaction for supplement intake occurred (P = 0.005) because HF steers were offered a constant amount of supplement daily, whereas steers fed AS, CAOH, and NACL were allowed to consume ad libitum quantities of supplement. Average daily gain in Trial 1 was not affected (P = 0.21) by supplementation. In Trial 2, NACL, AS, and HF treatments had higher (P < or = 0.07) ADG than CONT. In Trial 1, final weights were not affected by supplementation (P = 0.23). In Trial 2, final weights of NACL- and HF-fed steers were greater than for CONT and CAOH steers (P < 0.10). In Trial 2, CONT steer final weights were lower than all supplemented treatments (P < 0.10). For yearling steers grazing native range, use of NACL as a limiter resulted in increased weight gains compared with using either CAOH or AS; however, no limiter that was tested restricted supplement intake as effectively as HF. More research is necessary to determine the optimum limiter level and the effect of forage quality on supplement intake. PMID- 14753375 TI - Interactions between supplement energy source and tall fescue hay maturity on forage utilization by beef steers. AB - This experiment was conducted to determine the effects of tall fescue hay maturity on intake, digestion, and ruminal fermentation responses to different supplemental energy sources fed to beef steers. Twelve ruminally cannulated, crossbred steers (initial BW = 228 +/- 21 kg) were used in a split-plot experiment with a 3 x 4 factorial treatment arrangement. Steers were assigned randomly to three supplement treatments: 1) no supplement, 2) pelleted soybean hulls, or 3) coarse cracked corn. The second treatment factor was fescue hay maturity: 1) vegetative (VEG), 2) boot-stage (BOOT), 3) heading-stage (HEAD), and 4) mature (MAT). Supplements were fed once daily at 0.67% of BW (OM basis) and tall fescue hay was offered once daily at 150% of average intake. Supplement type x forage maturity interactions were not detected (P > or = 0.25) for forage, total, or digestible OM intake, which generally decreased (P < 0.01) with advancing forage maturity. Supplementation decreased (P < 0.01) forage and increased (P < 0.01) total OM intake. Supplement type had no effect (P = 0.56) on substitution ratio (unit change in forage intake per unit of supplement intake). Digestible OM intake was increased (P < 0.01) by supplementation and was greater (P = 0.05) with soybean hulls than with corn. Supplement type x forage maturity interactions (P < or = 0.10) were observed for OM and NDF digestibilities and N retention. Increases in digestibility with soybean hulls relative to corn were greater and supplementation elicited greater increases in N retention with more mature forages. Compared with soybean hulls, corn supplementation resulted in greater (P < 0.01) negative associative effects on OM digestibility. Supplementation did not affect (P > or = 0.10) ruminal pH, total VFA concentrations, or acetate:propionate ratio. Corn supplementation decreased (P < or = 0.07) ruminal NH3-N concentrations compared with control and soybean hulls; however, decreases in ruminal NH3-N concentrations were not consistent with the presence of negative associative effects. Thus, mechanisms not involving ruminal pH or NH3-N concentration seem responsible for negative associative effects observed with corn supplementation. Within the range of forage quality in this study, increases in digestible OM intake from starch- or fiber-based supplements were independent of forage maturity. When fed at similar levels of OM, soybean hull supplementation provided an average of 6% greater digestible OM intake than corn supplementation. PMID- 14753377 TI - CT imaging of patients with metallic hardware. PMID- 14753376 TI - Challenges and opportunities in the analysis of raffinose oligosaccharides, pentosans, phytate, and glucosinolates. AB - In this paper, the status of the analytical technologies for assaying animal antinutritional compounds, such as raffinose oligosaccharides, pentosans, phytic acid, and glucosinolates, is reviewed in terms of selectivity, sensitivity, and sample throughput. The implementation of simplified sample preparation schemes, use of novel separation approaches, and alternate detector technologies are discussed. The challenges and opportunities posed by these assays are highlighted along with the recommendations for best analytical practices. PMID- 14753378 TI - Case of the month. Diverticultitis with portomesenteric vein gas. PMID- 14753379 TI - Pulmonary alveolar proteinosis. PMID- 14753380 TI - Primary MALT-lymphoma of the liver: multimodality imaging. AB - MALT lymphoma rarely affects the liver. We present a case of primary MALT lymphoma of the liver, which appeared as multifocal hyperattenuated lesions compared to the fatty liver on unenhanced CT and as moderately hyperintense on T2 weighted and hypointense on T1-weighted MRI. We describe the radiological imaging features and discuss the differential diagnosis. PMID- 14753381 TI - Vaccine allergy: diagnosis and management. AB - As a group, vaccines provide a safe and effective way of preventing infectious and allergic illness. Allergic reactions to vaccines and drug products have become important and common features of practice and demand heightened awareness. Serious adverse effects of vaccines are rare but have been reported to various components of different vaccines. Although there are few precise diagnostic tests available, patients usually can be diagnosed accurately after careful attention to the history and physical findings. Better understanding of these reactions can lead to proper vaccine selection and can improve immunization acceptance rates in the community. Prevention, avoidance, use of alternative agents, desensitization, and premedication remain the mainstays of therapy, even as more refined diagnostic and management tools are developed. VAERS data, in addition to the traditional uses (signal detection, large registry of rare vaccine adverse events), can serve as a source of cases for epidemiologic (eg, case-control) studies that evaluate biologic factors that may be related to vaccine-related adverse reactions. Additional studies that are aimed at identifying other causes of immediate hypersensitivity after immunization with live virus vaccines are warranted. PMID- 14753382 TI - Vaccine safety. AB - Rates of reported adverse events are remarkably low. VAERS identifies an adverse event rate approximating 11.4 reports per 100,000 vaccine doses. Approximately 15% of these reports represent SAEs, but less than 2% involve death; in most cases, reviews have shown no causal relation between the events and the vaccine. Across the spectrum of vaccines in use (including those directed against influenza and hepatitis B virus), many claims of adverse events regarding vaccines represent typical reactions to vaccinations. These reactions can be thought of as foreign-body reactions and predominate among the inactivated vaccines. In controlled studies, the adverse event rates that occur with vaccination resemble those that occur with placebo injections. Typical reactions associated with live viral and bacterial vaccines, such as MMR and varicella vaccines, may resemble attenuated forms of the disease for which the vaccine is directed. Other claims against vaccines represent chance-coincidence or misunderstood data; further studies of claims have vindicated the overall safety of the vaccines in most cases. Two documented safety concerns with vaccines, however, have demonstrated that vaccines (like other biologics and pharmacologic) can result in harm (eg, rotavirus and OPV vaccines). The denouement with these vaccines indicates the broad postmarketing data collection and evaluation that extends efforts made with prelicensure study to balance the benefits from vaccination with the risk for harm. Overall, measures including prelicensure study and postlicensure surveillance, such as VAERS, the Vaccine Safety Datalink Project, and the Clinical Immunization Safety Assessment Centers, have resulted in an exceptional safety profile for the vaccines in use. PMID- 14753384 TI - Immunization of egg-allergic individuals with egg- or chicken-derived vaccines. AB - Viruses used in several vaccines are propagated in embryonated eggs. These vaccines contain variable quantities of residual egg or chicken proteins and pose risks when administered to egg- or chicken-sensitive persons. This article highlights differences in how vaccines are prepared, with emphasis on the quantitation of residual egg-derived protein in each vaccine. Published reports on the frequency and severity of these vaccine-induced allergic reactions are reviewed, and an algorithm is provided for the preimmunization evaluation of egg sensitive persons. PMID- 14753383 TI - Immunization of immunocompromised persons. AB - Advances in medicine, science, and technology have led to increasing numbers of people in the general population with altered host defenses. The risk for clinical infection in an immunocompromised host, such as a person who has received a solid organ transplant, is determined largely by the interaction between two factors: the epidemiologic exposures the person encounters and the person's net state of immunosuppresson. Vaccination represents a crucial approach for preventing infection in the general public and immunocompromised persons. This article reviews the benefits of and risks for immunization in immunocompromised persons and provides recommendations for the use of specific vaccines. PMID- 14753385 TI - Does antigenic overload exist? The role of multiple immunizations in infants. AB - There is no evidence that currently recommended vaccines overload or weaken the infant immune system. Infants have an enormous capacity to respond safely and effectively to multiple vaccines. The schedule for the administration of childhood vaccines is tailored to the unique developmental pattern of the infant immune system. Childhood vaccines provide immediate protection from common childhood illness and establish the foundation for lifelong immunity that develops with subsequent vaccination or infection. Widespread vaccination of infants and children represents a public health triumph of the 20th century. This fact must be reinforced continually by health care workers and parent education to help maintain progress in the 21st century. PMID- 14753386 TI - Allergic reactions to Japanese encephalitis vaccine. AB - The JEV widely is used in Asian countries each year and is an important vaccine for travelers to the East from other parts of the world. JE virus is a zoonotic disease with natural reservoirs and cannot be eliminated. Although a declining incidence of JE has been observed in Asia because of reduced transmission by agricultural approaches and vaccination, the most important control measure now, and in the future, is vaccination of humans against JE. The inactivated vaccine, produced from infected mouse-brain-derived tissue, is the only commercially available vaccine. There are several concerns with the use of this vaccine. It is expensive, requires two or three doses to achieve protective efficacy, and, in practice, requires further booster doses to maintain immunity. The apparent increase in allergic reactions in the first part of the 1990s has set focus on the safety of the JEV. A cheap, live attenuated SA 14-14-2 vaccine is used almost exclusively in China and parts of Korea, but there have been no trials of SA 14 14-2 vaccine outside JE endemic countries. The vaccine seems to be highly efficient, and few adverse events have been observed; however, PHK cells are used for the production of this vaccine, and these cells are not approved by the WHO. A satisfactory cell substrate is needed. A committee under the WHO has proposed that for the live JEV, there should be validity of the assays for retrovirus when applied to PHK cell substrate and validity of the mouse assays for neurovirulence. Further information should be reviewed on the long-term follow-up of recipients of the vaccine. Several new types of vaccines have reached the phase of clinical trials; however, studies remain to be completed. Until a new vaccine is available, the priority of surveillance of adverse events and the continuous reporting of such events to the users of the vaccines must be of importance. This fact is highlighted by the possibility of the varying frequency of adverse events with different batches over the years. The WHO offers information and recommendations for vaccines in the EPI and issues a series of updated papers on other vaccines that are of international public health importance (eg, JEV). The development of alternative efficient, safe, and appropriately priced JEVs is recommended, as is intensified surveillance of adverse events. Prospective vaccine studies of safety may be limited because of sample size and because rare adverse events may not be detected. Several new initiatives have been taken to improve surveillance of adverse events to vaccines within the past 10 years. In Japan, there is an increasing awareness of the importance of efforts taken to improve vaccine safety, and surveillance of adverse events and possibilities of compensation for vaccine-related injuries are in place. In Vietnam, a database to detect adverse events after vaccination has been established; the project involves active visits to data collectors at the vaccination sites. Comparative studies of adverse events, such as one recent study from Japan and the United States, are important for the evaluation of the reporting systems. The reporting rate for JEV adverse events from Japan was approximately one order of magnitude lower than that in the United States. Japan had strict predefined reporting criteria and time limits for observations. If time limits for the observation are too strict (eg, defining a possible neurologic reaction to occur within 1 week after vaccination), later reactions will not be included (eg, if ADEM is elicited by a vaccine, the symptoms cannot be expected to occur until weeks after the vaccination). The passive surveillance systems have limitations with an underreporting of adverse events, depending on clinical seriousness, temporal proximity to vaccination, awareness of healthcare workers, and tradition of reporting particular events. In developed countries, surveillance of adverse events is formalized, although not necessarily optimal. An increase in reporting would be expected when the reporting of adverse events is mandatory. Reports have been sent to VAERS, the Vaccine Safety Datalink Project, and the European Union Pharmacovigilance System. A Brighton collaboration has been implemented to enhance comparability of vaccine safety data. Public health authorities in specific countries, such as the CDC in the United States and the National Advisory Committee in Canada, regularly have published information on the JE situation in Asia and the preventive measures to be taken, including information on the vaccines and adverse reactions. The conventional recommendation is that travelers should be vaccinated if they will spend more than 1 month in a JE endemic area or in areas with epidemic transmission with even shorter periods. Although the risk for JE for short-term travelers is considered small (1 case per 1 million travelers per year), sporadic cases, including deaths, have been reported among tourists traveling to endemic areas. Risk for travelers in rural districts in the season of risk is considerably higher (range, 1 case per 5000 travelers to 1 case per 20,000 travelers per week). Doctors who advise travelers should be updated on the latest JE occurrences in Asia. Updates on the JE situation can be found on bulletins at http://www.promedmail.org or are available from the WHO or CDC. The allergic reactions primarily described after vaccination with the inactivated mouse-brain derived JEV have been observed in several countries during the 1900s. Allergic reactions, including the mucocutaneous and neurologic reactions reported after JE vaccination, may vary in frequency, and these reactions should be evaluated meticulously yearly. This step enables recommendations, including information on possible side effects, to be given in an optimal way. PMID- 14753387 TI - Aluminum inclusion macrophagic myofasciitis: a recently identified condition. AB - The authors conclude that the persistence of aluminum hydroxide at the site of intramuscular injection is a novel finding which has an exact significance that remains to be established fully. It seems mandatory to evaluate possible long term adverse effects induced by this compound, because this issue has not been addressed (in the past, aluminum hydroxide was believed to be cleared quickly from the body). If safety concerns about the long-term effects of aluminum hydroxide are confirmed, novel and alternative vaccine adjuvants to rescue vaccine-based strategies should be proposed to ensure the enormous benefit for public health that these vaccines provide worldwide. PMID- 14753388 TI - Anthrax vaccine: a review. AB - Anthrax can be a deadly disease if treatment does not begin early in the course of infection. An effective vaccine has been available in the United States since 1970, although it was not used widely until 1998. A comprehensive, peer-reviewed evaluation by the National Academy of Sciences affirmed the findings of multiple previous independent panels that found that the US-licensed anthrax vaccine is safe and effective. PMID- 14753389 TI - Smallpox vaccine: problems and prospects. AB - Smallpox justifiably is feared because of its morbidity and mortality. Wide spread population-level susceptibility to smallpox exists, and the only effective tool against the virus is a live, attenuated vaccine that is highly reactogenic and controversial. A significant minority of the population has contraindications that prevent preexposure use of this vaccine. Newer, safer, and equally immunogenic vaccines must be developed and licensed. Several live, attenuated vaccines are in clinical trials. Although these vaccines may prove to be less reactogenic, they still may not be administered safely to a significant portion of the population because they contain live, attenuated viruses. Newer vaccines will be needed if routine preexposure vaccination is to be instituted universally. The idea of a subunit or peptide-based vaccine is appealing, because it obviates potential safety concerns. It may be possible to use a more attenuated, live vaccine strain for a large segment of the population on a preexposure basis and accept the morbidity and mortality that would result from its use on a postexposure basis, if necessary. The need for widespread population level protection against variola infection is apparent. The use of the new biology tools to predict or define who might experience serious reactions to the smallpox vaccine and why these reactions occur is an area ripe for additional research. The reason why an individual develops postvaccinal encephalitis remains unknown, and the development is unpredictable and untreatable. In the future, if the mechanism behind such adverse events is defined, it may be possible to screen persons who are likely to experience such events. Although the authors remain proponents for use of the vaccine in alignment with the CDC vaccination program and recommendations, the previous concerns indicate that new knowledge must be gained and shared. Further research on attenuated vaccines and nonliving or peptide vaccines with equal efficacy should remain the goal, as it is apparent that smallpox vaccine once again will become part of the vaccinologist's and public health official's armamentarium in the decades to come. PMID- 14753390 TI - Safety and efficacy of live attenuated, cold-adapted, influenza vaccine trivalent. AB - This article describes the efficacy, immunogenicity, and safety of CAIV-T. This vaccine has the potential to significantly contribute to the control of influenza infection and influenza-associated illnesses, including febrile otitis media and lower respiratory disease. When compared with inactivated vaccine, CAIV-T has significant advantages in convenience of administration. The high efficacy of CAIV-T and its efficacy in children against a significantly drifted strain of H3N2 (A/Sydney), a strain not contained in the vaccine, are compelling observations for use of the vaccine in children. Effectiveness in adults was demonstrated using the same vaccine strain against the drifted H3N2 strain. The proposed vaccine administration schedule for healthy individuals aged 9 to 49 years is a single dose administered annually before the winter. For children aged 5 to 8 years, two doses are recommended the first year they are immunized with CAIV-T to ensure protection against all strains contained in the vaccine. Thereafter, a single annual revaccination is sufficient. PMID- 14753391 TI - Meningococcal immunology. AB - There is a major need for an effective vaccine against serogroup B disease. The long-term efficacy of the serogroups A, C, Y and W135 conjugate vaccines and the need for booster vaccines has to be determined, as does the effect of changing epidemiology in the United States and worldwide. Control of serogroup A disease in sub-Saharan Africa is a major challenge. PMID- 14753392 TI - Transcutaneous immunization and immunostimulant strategies. AB - The skin provides an attractive immune environment for vaccine delivery and a safe and confined anatomic space for the use of potent adjuvants. It has been presumed that LCs as a class of dendritic cells should stimulate potent immune responses when activated by adjuvants, and this theory is beginning to be validated. Progress on simple pretreatment of the skin has led to well-developed, simple-to-use protocols that are not dissimilar from current protocols used to cleanse the skin before injection. Antigen and adjuvant formulation optimization has progressed, leading to phase 2 testing of the technology in formulated, manufacturable patches. Although delivery optimization and product testing is challenging, the major biologic observations underlying TCI and the IS patch have been established clearly in that large protein antigens have been delivered clinically, resulting in robust immune responses in a safe manner. During the next 5 years, the challenge will be to conduct a development program that leads to safe and effective vaccination in the context of specific applications. PMID- 14753393 TI - Concept and pathogenesis of "hypoxic-ischemic encephalopathy". AB - By experiments of acute carbon-monoxide intoxication, acute nitrogen hypoxia and histotoxic hypoxia using sodium cyanide in cats, and by hemodynamic studies using plastic branch models, the following was elucidated; (1) severe tissue hypoxia, regardless of the underlying cause, and subsequent slight ischemia of the brain due to mild hypotension induce selective involvement of the cerebral white matter and pallidum, these two conditions being necessary and sufficient and this encephalopathy should be separately categorized as "hypoxic-ischemic encephalopathy" in hypoxic brain injuries, (2) the background of the selective involvement of these structures is an enormous development of the cerebrum in the brain, which induces thick white matter resulting in proper and long medullary artery, and especially small diameter ratio of the pallidal perforators to the middle cerebral artery, (3) the long course of the medullary artery produces the blood pressure drop in the deep white matter according to Hagen-Poiseuille's low, and according to that the smaller the diameter ratio, the larger the branching loss coefficient (energy-loss co-efficient), smaller diameter ratio of the pallidal perforator, as compared with that of the putaminal perforator, induces more severe loss of the local blood flow selectively to the pallidum. This state seems to be a failure of compromise between the cardiovascular system and the brain parenchyma. PMID- 14753394 TI - Pathophysiology of traumatic brain edema: current concepts. AB - The generally held concept during the past several decades is that traumatic brain edema is predominately vasogenic emanating from the blood vessels subsequent to blood brain barrier compromise. Much of the experimental data has focused on cryogenic injury models where there clearly is a necrotic lesion surrounded by leaking vessels. However, in closed head injury where brain swelling remains a critical problem, the classification of the type of edema that develops is less clear. Most importantly, studies in the clinical setting have ruled out vascular engorgement as one potential mechanism and these studies have shown that edema and not blood volume is the culprit responsible for brain swelling. We have put forth the notion that traumatic brain edema is a combination of vasogenic and cellular with the cellular component predominating. This article provides an update of our current progress toward supporting this hypothesis and includes an update on the role of aquaporins in traumatic brain edema. PMID- 14753395 TI - Brain edema from intracerebral hemorrhage. AB - Sequential changes in brain parenchyma surrounding an intracerebral hemorrhage are described here. Re-bleeding occurs within the first several hours after the initial hemorrhage in about 30%, of cases. The coagulation cascade is activated as soon as blood encounters tissue. Perihematomal brain edema develops in response to clot retraction, thrombin formation, erythrocyte lysis, hemoglobin toxicity, complement activation, mass effect, and blood-brain barrier disruption. Early hematoma evacuation interrupts edema formation. The toxicity ofextravasated blood in brain parenchyma has not been studied well in traumatic injury or in hemorrhagic tumor models yet, but similar mechanisms of edema formation are likely to occur in these conditions. PMID- 14753396 TI - Hydrostatic brain edema: basic mechanisms and clinical aspect. AB - The term hydrostatic brain edema results from unfavorable hydrtostatic pressure gradiernts between blood vessels and brain tissue. Arterial hypertension combined with decompressive craniectomy produces extensive brain edema in the arterial boundary zone of the decompressive area. The increased hydrostatic pressure gradient enhances tissue damage and causes the biphasic opening of the blood brain barrier (BBB). The hydrostatic pressure alone is capable of causing the initial BBB opening and this induced edema results in derangement of cerebral microcirculation and metabolism. With morphological opening and metabolic damage of the vascular wall, the second BBB opening is elicited by the amplified hydrostatic pressure gradient, which is similar to that in vasogenic edema. In the clinical aspect, the beneficial effect of decompressive craniectomy in the treatment of uncontrollable ICP and brain edema remains controversial. External decompression may have adverse effects on severe brain edema and swelling. In this regards, control of driving force for the formation of brain edema could be the treatment of choice as an initial step. In addition, recent reports provided by MR imaging indicated new information on the pathophysiological features of the patients with acute hypertension. In the patients with hypertension due to reversible posterior leucoencephalopathy (RPL) syndrome, MR images show reversible signal abnormalities in the bilateral occipital lobes, suggesting hydrostatic brain edema. PMID- 14753397 TI - Non-invasive imaging methods for the characterization of the pathophysiology of brain ischemia. AB - Non-invasive imaging methods are increasingly used to study the evolution and therapy of brain diseases under both clinical and experimental conditions. In the animal experiment, these methods can be supplemented by invasive tissue assays to allow precise characterization of the underlying pathophysiology. Based on such an approach, this review evaluates the importance of in vivo nuclear magnetic resonance (NMR) and positron emission tomography (PET) for the understanding of the pathophysiology of brain ischemia. PMID- 14753398 TI - Cecile & Oskar Vogt: the significance of their contributions in modern neuroscience. AB - A) By introducing and propagating the use of multidisciplinary approaches, Cecile and Okar Vogt played a decisive role in establishing brain research as a distinct branch of science, and thus laid the foundations for modern Neuroscience. B) The Vogts' and Brodmann's monumental work on brain cytoarchitectonics, supplemented with electrical stimulation of the exposed cerebral cortex, demonstrated correlations between evoked functional responses and the topography of brain regions endowed with distinct cytoarchitectonic structure (topistic units). This opened the field of functional brain mapping, a subject hotly pursued at the present time. C) One of the most important aspects of Pathoclisis is that the Vogts based their theory on molecular genetics. They envisaged that in any topistic unit the neurons had a different specific, molecular ("physico chemical") composition, accounting for differences observed in the dynamics of neuronal development and involution. In sublethal injury, the induced genomic reactivity varies according to the selective vulnerability of various topistic units. Thus, in cerebral ischemia, the fluctuating genomic reactivity may play a significant role in designing therapeutic strategies, which would include enhancing regenerative expressions and suppressing harmful ones. D) The pathoclisis theory may be of assistance in analyzing the nature of various genetic disorders, which could be considered for the application of gene therapy. PMID- 14753399 TI - Estimating blood-brain barrier opening in a rat model of hemorrhagic transformation with Patlak plots of Gd-DTPA contrast-enhanced MRI. AB - Patlak plot processing of Gd-shifted T1 relaxation-time images from a rat model of hemorrhagic transformation yielded estimates and maps of the blood-to-brain influx rate constant of Gd-DTPA (K1). The Patlak plots also produced a heretofore unrecognized parameter, the distribution space of the intravascular-Gd-shifted protons (Vp), an index of blood-to-tissue transfer of water. The K1 values for Gd DTPA were very high for the regions of blood-brain barrier (BBB) opening and were similar to those of 14C-sucrose concurrently obtained by quantitative autoradiographic (QAR) analysis. In these same ROI's, Vp was five-fold greater than normal, which suggests that the permeability of the BBB to water was also increased. The 14C-sucrose space of distribution in the ischemic ROI's was around 8%, thus indicating a sizable interstitial space. The spatial resolving power of Gd-DTPA-deltaT1 imaging was rather good, although no match for 14C-sucrose-QAR. This study shows that quantitative deltaT1-MRI estimates of regional blood-brain transfer constants of Gd-DTPA and water distribution are possible when Patlak plots are employed to process the data. This approach may be useful for tracking the time-course of BBB barrier function in both animals and humans. PMID- 14753400 TI - Recent development of MR imaging technique for the investigation of brain function. AB - Recently, a variety of MR techniques have been developed to investigate brain functions, such as high resolution brain images and various functional brain images. The BOLD fMRI technique opened the new field of non-invasively investigating local brain activities. As analyses are becoming more and more detailed, we frequently meet the problem of limitations with regard to S/N and the physiologic motions of the subjects to be measured. One solution for higher S/N is the introduction of high field MR systems. Improvement can also be achieved via the instrumental side. The Magnetom MR systems from Siemens introduced a high performance gradient system and flexible pulse sequence structure. Also great advances were made in fMRI measurements with real-time motion correction by introduction of PACE (prospective acquisition correction) technique. To apply such sophisticated technique in the clinical setting, the concept of in-line evaluation technique is employed, which means real-time data processing. As soon as scanning is terminated, the processed images can be obtained to be used immediately for diagnoses. PMID- 14753401 TI - Energy metabolism and cerebral blood flow during cytotoxic brain edema induced by 6-aminonicotinamide. AB - We investigated the progression of cytotoxic brain edema induced by 6 aminonicotinamide (6-ANA), a potent antimetabolite of nicotinamide, by measuring the time courses of changes in brain tissue water state (with MRI), histology (with H&E staining), energy metabolism (with 31P-NMR), brain hemoglobin concentration (with near-infrared spectroscopy; NIRS), cerebral blood flow and volume (CBF. CBV), mean arterial blood pressure (MABP), and brain activity (with EEG) up to 10 hours (h). Change in cerebrovascular autoregulation was also investigated. 6-ANA (120 mg/kg) was administered intraperitoneally to 30 male Wistar rats (250-350 g). After 10 h, the T2-weighted signal intensity was increased (p < 0.05), and H&E staining showed severe vacuolation of glial cells. ATP production/consumption and intracellular pH were well maintained up to 10 h, while the intensity of the phosphomonoesters (PME) signal was significantly increased (p < 0.05). Oxygen consumption gradually decreased from 4 to 10 h. CBF and MABP were all significantly increased (by 2.5-fold for CBF) (p < 0.05). Theta and delta wave amplitudes were reduced at 10 h. In summary, 6-ANA (120 mg/kg) induced cytotoxic brain edema from 4 to 10 h. Energy balance and brain activity were well maintained up to 10 h, though cerebrovascular autoregulation was impaired. PMID- 14753402 TI - PET neuroreceptor imaging as predictor of severe cerebral ischemic insult. AB - Measurement of the adenosine A1 receptor (A1-R) with positron emission tomography (PET) using a newly developed positron ligand, [1-methyl-11C]8 dicyclopropylmethyl-1-methyl-3-propylxanthine (MPDX). were performed in a cat middle cerebral artery (MCA) occlusion and reperfusion. Eighteen adult cats underwent PET measurement of; 1) cerebral blood flow (CBF). 2) A1-R, 3) central benzodiazepine receptor (BDZ-R) and 4) glucose metabolism with 15O labeled water, MPDX, 11C-flumazenil (FMZ) and 18F-fluorodeoxyglucose (FDG), respectively. The CBF, A1-R, BDZ-R and FDG uptake were serially measured after 60 min occlusion of MCA in this order. MPDX binding and FMZ binding, but not CBF and FDG uptake, were significantly reduced in the groups with severer ischemic insult than in the groups with no or milder insults. Of the two receptor ligands, the reduction rate of the MPDX binding to A1-Rs was larger in a group that caused fatal ischemic insult. The newly developed PET in vivo imaging technique using MPDX was suitable in evaluating the function of adenosine and A1-Rs in relation to cerebral ischemia. PMID- 14753403 TI - PET investigation of post-traumatic cerebral blood volume and blood flow. AB - Hemodynamic changes following traumatic brain injury (TBI) may reflect cellular damage leading to secondary injury. The purpose of this study was to investigate the regional hemodynamic parameters acutely after TBI among regions in and around contusions. Sixteen patients (11 male, 5 female) showing evidence of contusion on CT and 18 normal volunteers (12 male, 6 female) underwent positron emission tomography (PET) with O-15 CO and O-15 H2O to estimate cerebral blood volume (CBV) and cerebral blood flow (CBF), respectively. A flow to volume ratio (FVR = CBF/CBV) was also calculated as an index of vasodilatation. The hemodynamic parameters were compared among contusion, pericontusion, and remote areas. Globally, hemodynamic parameters did not differ between patients and normal volunteers, and did not correlate with intracranial pressure (ICP). Regionally, contusional and pericontusional areas showed significantly lower CBF and FVR compared with normal volunteers, while CBV did not differ significantly. The correlation between CBF and CBV was significant (r = 0.37, p < 0.01). Remote areas did not show a significant difference in any of the PET parameters. In conclusion, regional brain edema is likely to occur in contusion and pericontusion areas, while some of the contusional tissue may show vascular engorgement. PMID- 14753404 TI - Quantitative evaluation of cerebral vascular permeability using multi-slice dynamic CT. AB - We developed a method to measure regional permeability of cerebral capillary vessels, extracellular fluid space volume, and vascular bed volume using multi slice dynamic CT with iodinated contrast medium. We implemented the method as a novel software "BBB study", and evaluated it in four cases of brain tumor. The examination time was about 20 minutes, and the resulting functional maps had quality sufficient to distinguish area of irregular permeability enhancement. PMID- 14753406 TI - Functional brain imaging of optical topography. AB - Hitachi medical corporation developed and produced a new modality called Optical Topography System "ETG-100". This system uses near infrared light as in spectroscopy. ETG-100 measures the oxy hemoglobin, deoxy hemoglobin and total hemoglobin volume changes in cerebral cortex. This optical topography system has various features, i.e., displaying the hemoglobin volume change in 2 dimensional images, non-invasiveness, compactness, low cost and less patient restraint. This system is now being used for the research of higher-order brain functions, for example language function, language recognition. Research of brain function disease of new born babies, cerebral blood flow infarction detection are expected. PMID- 14753405 TI - Acetazolamide vasoreactivity evaluated by transcranial harmonic perfusion imaging: relationship with transcranial Doppler sonography and dynamic CT. AB - To establish the reliability and clinical significance of transcranial ultrasonic harmonic perfusion imaging (HPI), we evaluated HPI's relationships with transcranial Doppler (TCD) and with dynamic CT (DCT), during acetazolamide (ACZ) vasoreactivity tests. METHODS: The subjects were 12 neurological patients. Time averaged maximum velocity (TAVMX) in the middle (MCA) and posterior cerebral arteries was measured by TCD. Time-intensity (-density) curves of HPI (DCT) after bolus intravenous contrast injections were created in 3 regions of interest (ROI) on the axial plane involving the temporal lobe, basal ganglia, and thalamus on both sides. Assessments of vasoreactivity were based on comparisons conducted before and after ACZ administration in terms of: a) relative changes (%delta) of the TCD TAVMX, b) HPI contrast area enlargement, c) %delta of calculated cerebral blood volume and flow of the HPI and DCT. RESULTS: 1) TCD vasoreactivity decrease in the left MCA tended to correlate with lower frequency of HPI contrast area enlargement on the left side. 2) HPI and DCT vasoreactivity tended to be disturbed in the same side ROIs. CONCLUSIONS: Transcranial HPI achieves repeatable non-invasive bedside evaluation of cerebrovascular reserve capacity through qualitative and quantitative measurements of brain tissue perfusion, and will have clinical value in pathophysiological follow-up and therapeutic effectiveness determination of neurointensive care patients. PMID- 14753407 TI - Significant shrinkage of extracellular space during global cerebral ischemia: differences in gray and white matter ischemia. AB - Brain extracellular space (ECS) provides an important microenvironment for neurons and glial cells. In the present study, we investigated differences in ischemic changes of ECS in gray and white matter during global ischemia in cats (n = 8). Diffusion capacity of tetramethylammonium ion (TMA+, 74.1Da) was evaluated by measuring extracellular TMA+ concentration applied via microdialysis. In both gray and white matter, TMA+ concentration significantly increased within 10 minutes after the induction of ischemia, and sustained during 120 min of ischemia. The increase, however, was slower and smaller in white matter than in gray matter. In other two animals, volume fraction and tortuosity of ECS were determined by iontophoresis during global ischemia. After 30 min of ischemia induction, volume fraction was decreased and tortuosity was increased in both gray and white matter. In white matter, decrease in volume fraction and increase in tortuosity were smaller than in gray matter. The present study demonstrates that cerebral ischemia induces not only significant shrinkage of ECS volume but also restricts molecular diffusion within. Smaller changes of diffusion capacity within white matter ECS during ischemia may be relevant for lower ischemic vulnerability of the region. PMID- 14753408 TI - NTP and PCr responses to hypoxia by hypothermic and normothermic respiring, superfused, neonatal rat cerebrocortical slices: an NMR spectroscopy study at 14.1 Tesla. AB - Although mechanisms of hypothermic neuroprotection during oxygen deprivation have long been investigated, further characterizations of various molecular mechanisms are appropriate. Anticipating future studies of hypothermia and hypoxia/ischemia, we investigated the extent to which our ex vivo, NMR-based, superfused brain slice model might be helpful. (Slices are approximately 350 microm thick, with 18 slices per 8 mm NMR tube.) 31P NMR spectroscopic measurements were made of hypothermia-induced changes in high energy phosphates, while simultaneously monitoring and controlling tissue temperature, using 1H NMR, the high spectroscopic resolution available at 14.1 Tesla (600 MHz for protons), and a recently published protocol. NTP and PCr concentrations in healthy, well oxygenated slices decreased to (55 +/- 15)% and (66 +/- 30)% of their respective values at 28.0 degrees C when warmed to 38.0 degrees C, in approximate agreement with earlier in vivo studies by others. During 30 min hypoxia NTP and PCr decreased to non-observable values, regardless of temperature. After reoxygenation, NTP and PCr recoveries as percentages of respective prehypoxia values were (63% +/- 16%; 70%) +/- 5%) for hypothermic slices (28.0 degrees C), and (46% +/- 13%; 41% +/- hypothermic neuroprotection during oxygen deprivation in this model, which appears suitable for use in further studies. PMID- 14753409 TI - Matrix metalloproteinases are not involved in early brain edema formation after cardiac arrest in rats. AB - INTRODUCTION: Resuscitation from cardiac arrest (CA) often results in a poor neurological outcome possibly due to an incomplete understanding of the pathophysiology of brain injury following CA-induced global cerebral ischemia. Brain edema is an important manifestation after CA and is associated with significant morbidity and mortality. The matrix metalloproteinases (MMPs) contribute to brain edema formation following focal cerebral ischemia. The objective of this study was to investigate the role of an MMP inhibitor, GM6001, in CA-elicited brain edema. METHODS: Eighteen rats were subjected to normothermic (37.5 +/- 0.5 degrees C) CA induced by eight minutes of asphyxiation and assigned to a CA-control group (CA), an alcohol-placebo group (CA + ETOH), or a GM6001 treated group (CA + GM6001). GM6001 in 100% alcohol or a vehicle was given i.v. before CA to achieve a whole blood concentration of 10 microM. Animals were resuscitated with CPR, ventilation and epinephrine. Brain edema was determined by brain wet-to-dry weight ratio at one hour after resuscitation. FINDINGS: Brain wet-to-dry weight ratio was 4.86 +/- 0.09 in CA, 4.76 +/- 0.12 in CA + ETOH (p = 0.30 vs. CA), and 4.72 +/- 0.03 in CA + GM6001 (p = 0.17 vs. CA and 0.42 vs. CA + ETOH). INTERPRETATION: MMPs are not involved in brain edema formation one hour following CA. PMID- 14753410 TI - Regional N-acetyl-aspartate level and immunohistochemical damage in the hippocampus after transient forebrain ischemia in gerbils. AB - We investigated changes in regional N-acetyl-aspartate (NAA) levels in the vulnerable CA1 and resistant CA3 areas of the hippocampus after transient forebrain ischemia in gerbils. Under light ether anesthesia, bilateral common carotid arteries of adult male Mongolian gerbils (60-80 g) were occluded for 5 min and reperfused for 7 days. Brains from experimental and control gerbils (n = 4 each) were frozen in situ, and frozen sections (20 microm) were prepared using cryostat (-20 degrees C). After overnight lyophilization, the CA1 and CA3 areas were dissected out separately, weighed (50-200 microg), and the supernatant of the perchloric acid extract was used for assay of NAA using HPLC. Adjacent 10 microm-thick sections were used for immunohistochemical analysis using antiserum against microtubule-associated protein I and II. The preischemic NAA levels were not significantly different between CA1 and CA3 areas. After transient ischemia, a significant (P < 0.01) decrease in the NAA level was observed in the CAI area, but not in the CA3 area of the hippocampus. Immunohistochemical ischemic damage evolved only in the CA1 area. Thus, the decrease of the regional NAA level was associated with development of immunohistochemical neuronal damage. PMID- 14753411 TI - Extracellular superoxide dismutase following cerebral ischemia in mice. AB - We describe the changes in extracellular superoxide dismutase (EC-SOD) following cerebral ischemia in mice. Mice were subjected to transient forebrain ischemia and reperfusion. The measurements of EC-SOD using ELISA showed increased brain EC SOD after 24 h of reperfusion. The immunohistochemical examination showed that EC SOD immunoreactivity in cortical and striatal capillary wall was conspicuous after 3 h. EC-SOD immunoreactivity was also noted in cortical neurons after 24 h. Northern blot analysis showed an increased EC-SOD mRNA expression in the brain after 24 h. In situ hybridization study demonstrated no mRNA expression of EC-SOD following ischemia and reperfusion in the capillary wall. These findings suggest that serum EC-SOD might accumulate on brain endothelial cells, while cortical neurons produce EC-SOD themselves after cerebral ischemia with reperfusion. PMID- 14753412 TI - Peroxynitrite and caspase-3 expression after ischemia/reperfusion in mouse cardiac arrest model. AB - NO is a putative neurotransmitter and neuromodulator in the brain. NO is not functioning as a direct neurotoxin. NO with the superoxide radical product peroxynitrite (ONOO-) is much more cytotoxic under tissue impairment conditions. Caspase-3, a potent effector of apoptosis that is triggered via several different signaling pathways, may play a very important role in neuronal cell death caused by various brain injuries. The relationship between mouse caspase-3 and peroxynitrite remains unclear. In the present study, we examined the in vivo expression of 3-nitrotyrosine (a metabolite of peroxinitrite) and caspase-3 after cerebral ischemia produced in a global ischemia model using mice (i.e., a cardiac arrest model). 3-nitrotyrosine immunoreactivity was detected in neuronal cells in the hippocampal dentate nucleus, and cortical regions starting at 12 hrs after ischemia. In particular, numerous neuronal cells were highly immunoreactive for 3 nitrotyrosine in the cortical regions. In hippocampal CA1 pyramidal neurons, 3 nitrotyrosine immunoreactivity was detected from 24 hrs. Caspase-3 immunopositive cells were observed in approximately the same area in which the positive reaction to the anti-nitrotyrosine antibody was observed. These results provide direct evidence for the induction of 3-nitrotyrosine and caspase-3 expression in vivo in an ischemia model using mice. The present findings suggest that peroxynitrite generated by cerebral ischemia/ reperfusion was strongly cytotoxic and induced neuronal cell death (apoptosis) mediated by caspase-3. PMID- 14753413 TI - Expression of tumor necrosis factor alpha (TNFalpha) following transient cerebral ischemia. AB - It has been considered that tumor necrosis factor alpha (TNFalpha) is participated in the Alzheimer's, and Parkinson's diseases, brain injury and brain ischemia. However, expression of TNFalpha after brain ischemia has not been demonstrated in detail. Therefore we examined the cellular expression of TNFalpha during and after transient middle cerebral artery occlusion (tMCAO) in mice by use of reverse transcriptase-polymerase chain reaction and immunohistochemical technique. TNFalpha mRNA expression was gradually increased in the neocortex of the ipsilateral hemisphere during ischemia and peaked at 1 hour after reperfusion. Then, the mRNA expression decreased and peaked again at 24 hours after reperfusion. TNFalpha-like immunoreactivities were observed in the process such as dendrite of neuron slightly before ischemia, and markedly increased in neurons in addition to the process of the ipsilateral hemisphere at 1 and 24 hours after ischemia. The results suggest that the expression of TNFalpha is up regulated in the neurons after tMCAO. TNFalpha may induce ischemic neuronal cell death during ischemic insult. PMID- 14753414 TI - Evaluation of neuronal cell death after a new global ischemia model in infant mice. AB - For the first time we set up a new model for global ischemia in the infant mice, and time-dependent changes of the blood-brain barrier (BBB) disruption and neuronal cell death were investigated in detail. Infant C57/B16 mice (postnatal 13 days) were anesthetized with inhalation of sevoflurane in N2O/O2 (70/30%) and were subjected to global ischemia by bilateral common carotid artery occlusion (CCAO) for 25 minutes. Disruption of BBB was noted at 4 hours and increased up to 24 hours after the injection of 2% Evan's Blue in the transient CCAO (tCCAO) model. Evaluation of neuronal cell death was determined with toluidine blue staining. Morphological changes of neurons after tCCAO were clearly observed in the hippocampal CA1 region but were slightly detected in the CA3 region. However, there were no morphological changes in the hippocampal dentate gyrus, the neocortex, the striatum and the hypothalamus. The number of survival neurons in the CA1 was significantly decreased at 2 days and sustained up to 4 days after tCCAO. These data indicate that this method is very useful to induce selective vulnerability in mouse hippocampus, and it provides a reliable ischemic model in infant mice. PMID- 14753415 TI - Hippocampal heme oxigenase-1 in a murine cardiac arrest model. AB - Heme oxygenase-1 (HO-1) is a member of the heat shock protein family (HSP-32). It responds to thermal stress in cultured glial cells. To our knowledge. nothing is known about the expression and response of the HO-1 in cerebral ischemia. Therefore, we show here the induction of HO-1 in the brain of mice after global cerebral ischemia. HO-1-like immunoreactivity was detected at 12, 24, and 48 hours after ischemia recirculation. The HO-1-like immunoreactive cells were observed in astrocytes in the hippocampal dentate gyrus and CA1. The peak level of HO-1-like immunoreactivity was found 48 hours after the recirculation. HO-1 like immunoreactivity was observed in GFAP-positive astrocytes by use of a double immunostaining method. These results provide direct evidence for the induction and localization of HO-1 immunoreactivity in vivo in a mouse cerebral ischemia. We suggest that HO-1, produced in astrocytes after ischemia-recirculation, may directly affect neurons to protect from cell death. PMID- 14753416 TI - Calcineurin and cyclophilin D are differential targets of neuroprotection by immunosuppressants CsA and FK506 in ischemic brain damage. AB - The search for an effective treatment for global ischemia following cardiac arrest has proved to be very difficult. However, studies by Uchino et al. show that the immunosuppressant cyclosporin A (CsA), when administered in such a way that the drug can bypass the blood brain barrier (BBB), dramatically reduces ischemic damage in rat forebrain preparations. An alternative immunosuppressant, FK506, is apparently less efficacious. Both CsA and FK506 are specific inhibitors of immunophilins, (CsA inhibits cyclophilins, FK506 inhibits FKBPs), and of calcineurin, a type 2B Ser/Thr phosphatase that is abundant in the central nervous system. The superiority of CsA may be partly attributable to its selective amelioration of mitochondrial damage, as assayed in vivo and in vitro. Our results suggest that pathways involving calcineurin and cyclophilins, particularly mitochondrial cyclophilin D, play pivotal roles in the development of ischemic brain damage. The present findings may inform the search for new drugs in the treatment of global ischemic damage to the brain, and in other organs. PMID- 14753417 TI - FK506 attenuates the post-ischemic perturbation of protein kinases and tyrosine phosphorylation in the gerbil hippocampal CA1 sectors. AB - To explore effects of Immunosuppressant FK506 on signal transduction pathway. we studied changes in subcellular distribution of protein kinase Cgamma (PKCgamma), CaM kinase II (CaMKII), as well as changes of tyrosine phosphorylation levels after ischemia. Male Mongolian gerbils were divided into 3 groups; FK506 (1 mg/kg, 3 mg/kg) and vehicle. FK506 was administered intravenously after 5 min ischemia. At the designated time points (0 time, 5 min ischemia, 1 hour, or 24 hour recovery), heads were frozen and samples were taken from CAI subfield of hippocampus. Western blot analysis was carried out with specific antibodies for PKCgamma, CaMKII, and phosphotyrosine. FK506 administration significantly decreased translocation of PKCgamma and CaMKII at 24 h of recovery (p < 0.05, ANOVA followed by Student-Newman Keuls' test) in P2 fraction. The levels of tyrosine phosphorylated p160, p140, p100, p90, and p80 in P2 fraction were also significantly decreased with FK506 treatment at 24 h of recovery. The persistently elevated PKCgamma and CaMKII level in P2 fraction which may be related to cell death, are attenuated with FK506 treatment. FK506 may contribute to recover calcium homeostasis in the post ischemic phase and promote cell survival. PMID- 14753418 TI - Blood-brain barrier dysfunction and amyloid precursor protein accumulation in microvascular compartment following ischemia-reperfusion brain injury with 1-year survival. AB - This study examined the late microvascular consequences of brain ischemia due to cardiac arrest in rats. In reacted vibratome sections scattered foci of extravasated horseradish peroxidase were noted throughout the brain and did not appear to be restricted to any specific area of brain. Ultrastructural investigation of leaky sites frequently presented platelets adhering to the endothelium of venules and capillaries. Endothelial cells demonstrated pathological changes with evidence of perivascular astrocytic swelling. At the same time, we noted C-terminal of amyloid precursor protein/beta-amyloid peptide (CAPP/betaA) deposits in cerebral blood vessels, with a halo of CAPP/betaA immunoreactivity in the surrounding parenchyma suggested diffusion of CAPP/betaA out of the vascular compartment. Changes predominated in the hippocampus, cerebral and entorhinal cortex, corpus callosum, thalamus, basal ganglia and around the lateral ventricles. These data implicate delayed abnormal endothelial function of vessels following ischemia-reperfusion brain injury as a primary event in the pathogenesis of the recurrent cerebral infarction. PMID- 14753419 TI - Neuroprotective effect from ischemia and direct free radical scavenging activity of Choto-san (kampo medicine). AB - Choto-san is a kampo medicine that is widely used in patients with cerebral infarction, but the details of its mechanism of action remain unclear. We examined the neuroprotective effects of Choto-san using an experimental cerebral ischemia model (i.e., a rat cardiac arrest model). We also investigated the ability of Choto-san to eliminate or inhibit the activity of free radicals. It was found that Choto-san significantly prevents delayed neuronal cell death after ischemic reperfusion. Electron spin resonance demonstrated that the formation of hydroxyl- and superoxide-DMPO spin adducts were inhibited by Choto-san. The results of this study indicated that Choto-san prevents delayed neuronal cell death in the hippocampal CA1 region after ischemia. Direct free radical scavenging activity is among the pharmacological effects of Choto-san. PMID- 14753420 TI - Temporal profile of experimental ischemic edema after threshold amount of insult to induce infarction--ultrastructure, gravimetry and Evans' blue extravasation. AB - When a threshold amount of temporary ischemic insult to induce focal infarction was given to the unilateral cerebral hemisphere of gerbils, a small focal infarct surrounded by a wide penumbra developed in the rostral portion of the cerebral cortex. During the first 5 hours following recirculation, whole astrocytic cell bodies and processes in the ischemic hemisphere were swollen, with an increase in the number of glycogen granules and in number and size of mitochondria. This swelling was an active reaction of astrocytes for neuronal protection, scavenging potassium, glutamate, and other neuronal metabolic products, and for generating fuels for neurons (cyto-reactive edema). This reactive astrocytic swelling continued in the penumbra, but some dead neurons were found disseminated among the surviving neurons. Whereas, at 12 approximately 48 hours, focal infarction developed in which all cell membranes lost their Gibbs-Donnan's equilibrium due to energetic failure of their membranous Na+/K+ ATPase. This is the cytotoxic edema (cyto-necrotic edema). In the infarct focus, when pericapillary astrocytic end-feet were damaged, the capillary BBB was broken; and thus vasogenic edema was superimposed on the cytotoxic edema. PMID- 14753421 TI - Prediction of tissue survival after stroke based on changes in the apparent diffusion of water (cytotoxic edema). AB - This study tested the hypothesis that the estimate of the apparent diffusion coefficient (ADC; an index of cytotoxic edema) of water is a reliable pathophysiological index of the viability of ischemic brain tissue. CBF, the partition volume of water (PVW; an index of vasogenic edema), cerebral metabolic rate of oxygen (CMRO2), and of glucose (CMR(glc)) were measured before and after permanent middle cerebral artery occlusion (MCAO) or reperfusion with positron emission tomography (PET) in pigs. Then, the ADC was measured by diffusion weighted magnetic resonance imaging (DW-MRI) and was compared with physiological variables obtained by PET and with histological findings. Both after permanent MCAO and reperfusion, the ADC was significantly correlated to the CMRO2 and CMR(glc). The sequential decrease of ADC was also correlated to the sequential reduction of CMRO2 in relation to the residual CBF after permanent MCAO. The infarction coincided with a threshold CMRO2 of 50% of the contralateral value and a threshold ADC of 75% of the contralateral value, respectively. The PVW was decreased in relation to the magnitude of residual CBF after MCAO, whereas it was transiently increased in the severely ischemic lesions below 75% of the contralateral ADC value after reperfusion. Thus, the ADC is a reliable pathophysiological index which allows therapy to be tailored to the condition of individual patients in clinical practice. PMID- 14753422 TI - Recovery of apparent diffusion coefficient after embolic stroke does not signify complete salvage of post-ischemic neuronal tissue. AB - BACKGROUND AND PURPOSE: It has been shown in previous studies that recovery of apparent diffusion coefficient (ADC) of water is not necessarily associated with a reversal of tissue damage. We examined the relationships between temporal ADC change and histological outcome after embolic stroke in rat using an experimental MRI scanner. SUBJECTS AND METHODS: Male Wistar rats underwent embolization of the right middle cerebral artery with autologous clot. ADC and relative cerebral blood flow (rCBF) were measured consecutively for 3 hours after embolization. The brains were prepared for histological examination. RESULTS: Transient decline of ADC was observed during the initial 2 hours after embolization. In these areas, serial rCBF measurement revealed incomplete recovery. In some other areas, ADC decrease without rCBF recovery was observed during 3 hours postischemia. Histological examination revealed infarction in areas with persistent ADC decrease. Scattered neuronal death was noticeable in areas with a transient--in some areas less than 0.5 hour--ADC decrease. CONCLUSION: Consecutive MRI analysis demonstrated postischemic transient ADC decrease in an embolic stroke model of rat. The normalization of ADC does not signify normalization of postischemic tissue, since the areas often evolve postischemic neuronal death at a later time. PMID- 14753423 TI - Temporal evolution of apparent diffusion coefficient and T2 value following transient focal cerebral ischemia in gerbils. AB - We examined the time course of apparent diffusion coefficient (ADC) and T2 values in gerbils subjected to transient focal cerebral ischemia and compared them with histopathologic changes. Ten gerbils were subjected to two times 10-min occlusions of the left common carotid artery and examined with diffusion-weighted and T2-weighted MR imaging, at the interocclusion period and 1 hour, 2 hours. 1 day, 2 days, 4 days, and 7 days after the second occlusion. Immediately after the last MR imaging, their brains were examined histopathologically. ADC values decreased 1 hour after the second occlusion and continued to decrease up to 1 or 2 days later. ADC values remained lowered up to 4 days and slightly recovered at 7 days. T2 values were normal at 1 and 2 hours and began to increase at 1 day. T2 values began to recover at 4 days. Histopathologically, infarction was confined in the fronto-parietal cortex, dorsolateral caudate nucleus and dorsolateral thalamus, and neuronal necrosis was found in the pyramidal cell layer of the hippocampus. This study indicates that sequential ADC and T2 studies of gerbils subjected to transient focal cerebral ischemia provide a useful tool for evaluating temporal evolution of ischemic brain injury and edema, including cytotoxic and vasogenic edema. PMID- 14753424 TI - Prediction of malignant infarction: perifocal neurochemical monitoring following prolonged MCA occlusion in cats. AB - Neurochemical monitoring in the ischemic core predicts malignancy in focal ischemia in cats. Since perifocal regions are more suitable for clinical microdialysis (MD) applications, we tested whether monitoring at such site predicts also malignancy.--Laser Doppler (LD) probes, pressure microsensors, and MD/HPLC measured cerebral blood flow (CBF), intracranial pressure (ICP), and extracellular glutamate (Glu), respectively. The middle cerebral artery was occluded (MCAO) for 3 hours followed by 6 hours reperfusion. Additionally, LD measured CBF in ischemic core.--MCAO reduced CBF in the core in all below 25% of control. In animals exhibiting malignancy (eye dilatation during reperfusion), MCAO decreased CBF in the perifocal site to around 35%. CBF primarily recovered following recirculation but decreased thereafter as ICP rose due to progressive edema formation. Glu increased concomitantly. In cats exhibiting a benign course, MCAO decreased CBF in the perifocal site to around 55%. Recirculation normalized CBF, and Glu did not increase. During MCAO, Glu differences between both groups were not significant.--Glu determinations in perifocal sites taken during MCAO do not predict fatal outcome. This contrasts with determinations in the core. After reperfusion, Glu elevation in perifocal sites may serve as a rather late predictor of malignancy. PMID- 14753425 TI - A comparison of long-term neurological symptoms after two different focal ischemic models in Mongolian gerbils. AB - The aim of the present study was to elucidate the relationship between specific tasks and the responsible ischemic lesions after experimentally induced cerebral hemispheric ischemia in Mongolian gerbils. We used the elevated body swing test (EBST) to evaluate asymmetry motor behavior, the bilateral asymmetry test (BAT) to evaluate sensory dysfunction, and the T-maze test to assess cognitive deficit during 4 weeks after either 10-min single unilateral carotid artery occlusion (sCCAO) or 10-min repeated unilateral occlusion (rCCAO). rCCAO produced persistent sensorimotor and severe cognitive deficits. Infarction was confined to the ipsilateral cerebral cortex, hippocampus, dorsolateral nucleus of thalamus and caudate nucleus. sCCAO did not induce sensorimotor abnormalities, but did produce mild cognitive deficits; these animals also showed increased locomotor activity during the early post-ischemic period. In sCCAO, neuronal death was confined to the ipsilateral CA1 sector of the hippocampus. Thus, unilateral CA1 neuronal death was sufficient for evolution of cognitive deficits in this model of experimental focal ischemia. Ischemic lesions extending to the ipsilateral cerebral cortex, basal ganglia, and hippocampus produced persistent and severe sensorimotor impairment accompanying severe cognitive deficits. These findings regarding region-specific behavioral tasks in cerebral ischemia will facilitate improved assessment of stroke therapy. PMID- 14753426 TI - Thrombin exacerbates brain edema in focal cerebral ischemia. AB - Thrombin contributes to edema formation after intracerebral hemorrhage. Recent studies suggest that thrombin may also play a role in ischemic brain damage. In the present study, adult male Sprague-Dawley rats were anesthetized with pentobarbital. Middle cerebral artery (MCA) was occluded using the suture method. We found that brain thrombin activity was elevated after permanent MCA occlusion as was prothrombin messenger RNA expression. Intracerebral injection of a thrombin inhibitor, hirudin, reduced neurological deficits following cerebral ischemia. In contrast, intracerebral administration of exogenous thrombin (at a dose that is non-toxic to normal brain), markedly exacerbated brain edema after transient focal cerebral ischemia. These results indicate that extravascular thrombin inhibition may be a new therapeutic target for cerebral ischemia. PMID- 14753427 TI - Infiltration of tissue plasminogen activator through cerebral vessels: evaluation using a rat thromboembolic stroke model. AB - In recent years, evidence has been accumulating that tissue plasminogen activator (tPA) possesses neurotoxic effects. However, such deleterious effects have been attributed to endogenously generated tPA. In the present study, focusing on exogenously administered tPA for the purpose of fibrinolysis, we evaluated the extent and the degree of extravasated tPA in a rat model of thromboembolic stroke. Even after early recanalization of occluded cerebral vessels, significant infiltration of tPA occurred through the cerebral vessels. It is assumed that exogenous tPA also exerts neurotoxic effects in the ischemic brain tissue. PMID- 14753428 TI - Mice deficient in cytosolic phospholipase A2 are less susceptible to cerebral ischemia/reperfusion injury. AB - To determine the role of cytosolic phospholipase A2 (cPLA2) in infarct development, wild-type and cPLA2 knock-out mice were subjected to focal cerebral ischemia for 75 min by occluding the middle cerebral artery using nylon filament and subsequent reperfusion by withdrawing the filament. The neurological deficit severity was evaluated by a modified 4-point scale. After the reperfusion period (72 h), mice were killed, and the brains were cut into four 2 mm coronal sections using a rodent brain matrix. Sections were stained with 2% 2,3,5 triphenyltetrazolium chloride (TTC). The infarct volume was 87.19 +/- 27.54 mm3 (mean +/- SD, n = 11) in the wild-type mice and 48.20 +/- 31.32 mm3 (n = 10; P < 0.01 vs. wild-type) in the knock-out mice. Less severe functional neurological deficits were observed in knock-out mice at 72 h after ischemia when compared with wild-type. Thus, disruption of cPLA2 resulted in significant reduction of infarct area and neurological deficit severity in the MCA occlusion model. These data indicate a critical role for cPLA2 in the pathogenesis of cerebral ischemia/ reperfusion injury. PMID- 14753429 TI - Regional distribution of potassium and phosphorus in ischemic brain tissue of rats with X-ray fluorescence analysis. AB - Focal cerebral ischemia was induced by occlusion of the left middle cerebral artery (MCAO) for 1 and 3 hours in 14 rats. Five sham-operated rats served as controls. Regional cerebral blood flow (rCBF) was assessed by 14C-iodoantipyrine radioluminography. The potassium and phosphorus contents in brain tissue were assessed using energy-dispersive X-ray fluorescence spectrometry. In the inner periphery within the ischemic tissue where rCBF was severely reduced to 6.61 +/- 0.95 ml/100 g/min 1 hour after MCAO, the potassium content decreased slightly to 12.82 +/- 0.15 mg/g. Potassium content decreased further to 10.61 +/- 0.24 mg/g over 3 hours after occlusion, similar to that in the ischemic core. In contrast, in the outer periphery where rCBF was moderately reduced, the potassium fell slightly to 12.78 +/- 0.46 mg/g 3 hours after MCAO. Tissue phosphorus content decreased to 8.28 +/- 0.24 mg/g only in the ischemic core. These findings demonstrate that potassium and phosphorus in brain tissue depend primarily on the extent, magnitude and duration of ischemia. Ion homeostasis of cell membranes may not be completely disturbed in the periphery around ischemic tissue for a short time after the onset of ischemia, even though blood flow is markedly reduced. PMID- 14753430 TI - Changes of free cholesterol and neutral lipids after transient focal brain ischemia in rats. AB - We investigated temporal and spatial changes of free cholesterol (FC) and neutral lipids (NLs) after brain ischemia with filipin complex staining to detect mainly FC and Nile Red staining for NLs such as cholesteryl ester (CE) and triacylglyceride (TAG). Filipin stainining decreased during 1 to 7 d and lost at 21 d after transient middle cerebral artery (MCA) occlusion in the ischemic core, but did not change in the penumbra. Nile Red positive droplets reached the maximum at 7 d after transient MCA occlusion (MCAO) and gradually decreased in the core, while the peak time delayed in the penumbra. Most Nile Red positive droplets were double positive for ED1 in the core, and were localized within GFAP positive cells in the penumbra. The present study suggests that changes of FC and NLs are different temporally and spatially between the ischemic core and the penumbra in relation to degenerative and regenerative neural processes following brain ischemia. Macrophages and astrocytes are presumed to play important roles in lipid metabolism for neural reorganization of the ischemic brain injury in the ischemic core and the penumbra, respectively. PMID- 14753431 TI - Osteopontin infusion into normal adult rat brain fails to increase cell proliferation in dentate gyrus and subventricular zone. AB - In the first week after focal ischemia in adult brain, the basal level of neurogenesis increases dramatically in two distinct areas: The dentate gyrus (DG) of the hippocampus and the subventricular zone (SVZ) of the lateral ventricles. It is possible that this remotely induced neurogenesis is the result of a proliferation inducing factor, or factors, diffusing from the infarction to the neurogenic regions. The secreted protein osteopontin (OPN) is a possible factor. In this study, OPN mRNA levels were measured in the cerebral infarction of adult rats that underwent I hour of middle cerebral artery occlusion (MCAO). OPN mRNA levels increased 36.0, 55.0 and 46.7 fold at 6, 24 and 72 hours reperfusion respectively. We also determined whether OPN alone could be responsible for this ischemia-induced neurogenesis. OPN (2.4 microg/day) was infused into the lateral ventricles of the brain in non-ischemic adult male rats, continuously over three days. Bromodeoxyuridine (BrdU) immunohistochemistry was performed and the total BrdU positive (BrdU+) cells were counted. OPN, compared to aCSF infusion, decreased BrdU+ cells in DG and had no significant effect on cell proliferation in the SVZ. This study indicates that osteopontin alone does not increase cell proliferation in the normal adult brain. PMID- 14753432 TI - Dissociative increase of oligodendrocyte progenitor cells between young and aged rats after transient cerebral ischemia. AB - Oligodendrocyte progenitor cells (OPCs) mature to oligodendrocyte and remyelinate axons. OPCs express neuron-glial antigen 2 (NG2) chondroitin sulfate proteoglycan. We stained NG2 to investigate the effect of aging on the OPCs after cerebral ischemia. NG2 positive cells were examined at 1, 3, 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. At 7 day, 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 more highly branched in the young than the aged brains. These results suggest that the young brain shows a higher potential of proliferation and process branching of OPCs than the aged brain. PMID- 14753433 TI - Suppression of oxidative stress after transient focal ischemia in interleukin-1 knock out mice. AB - Interleukin-1 (IL-1) contributes to ischemic neurodegeneration. However, the mechanisms regulating action of IL-1 are still poorly understood. In order to clarify this central issue, mice that were gene deficient both IL-1alpha and beta (IL-1 KO) and wild-type mice were subjected to 1 hour transient middle cerebral artery occlusion (tMCAO). The concentration of 8-hydroxy deoxyguanosine (8OHdG) which is considered to be a reliable oxidative DNA damage by superoxide anion, in brain and of total nitric oxide (NO) in plasma were determined by use of HPLC. Twenty-four hours after tMCAO, the ratio of 8OHdG to dG in the ipsilateral hemisphere of wild-type mice were 2.24 x 10(-3) and 4.41 x 10(-3) in the neocortex and striatum, respectively. The concentration of 8OHdG in the ipsilateral hemisphere of the wild-type mice was higher than that of the IL-1 KO mice. The concentration of total NO in the plasma of IL-1 KO mice was also lower than that of the wild-type 24 hours after tMCAO. These results strongly suggest that IL-1 is participated in generating reactive oxygen spices and it aggravates and induces the ischemic neuronal cell death.(183 words). PMID- 14753434 TI - Mild hypothermia enhances the neuroprotective effects of a selective thrombin inhibitor following transient focal ischemia in rats. AB - The aim of this study is to determine whether a selective thrombin inhibitor, Argatroban, would prevent neuronal cell death and whether extra-mild hypothermia (35 degrees C) would enhance the neuroprotective effect of a selective thrombin inhibitor following transient focal ischemia in rats. Sprague-Dawley rats were subjected to MCAo using an intraluminal suture technique for 2 hrs. The rats were reperfused for 24 h and decapitated for infarct and edema analysis. Argatroban treated animals received a continuous injection of argatroban (3.0 mg/kg) for 24 hrs after onset of ischemia, while vehicle-treated groups received same dose of vehicle. During ischemia, temporal muscle and rectal temperatures were monitored and maintained at 37 degrees C in the normothermic animals and at 35 degrees C in the hypothermic animals. Argatroban ameliorated the cortical ischemic damage significantly (p < 0.05). Moreover, argatroban with mild hypothermia decreased the cortical infarct or edema volume significantly compared with those of groups I and III (p < 0.05). Argatroban improved neurological symptoms significantly and also improved survival rate. These results demonstrate that extra-mild hypothermia (35 degrees C) enhances neuroprotective effects of a selective thrombin inhibitor, argatroban, suggesting that this combined therapy may be a new therapeutic strategy for the treatment of acute stroke. PMID- 14753435 TI - The neuroprotective effect of a free radical scavenger and mild hypothermia following transient focal ischemia in rats. AB - Edaravone, a novel free radical scavenger, has been reported to reduce ischemic damage in rats subjected to transient focal ischemia. The aim of this study is, therefore, to investigate the effect of a combined therapy with edaravone and mild hypothermia of 35 degrees C. Sprague-Dawley rats were subjected to MCA occluding an intraluminal suture technique for 2 hrs. The rats were reperfused for 24 h and decapitated for infarct and edema analysis. Animals were randomly devided into four groups: (I) vehicle + normothermia (control) (II) vehicle + mild hypothermia (III) Edaravone + normothermia (IV) Edaravone + mild hypothermia. Mild hypothermia alone had no reduction of the brain damage. The edaravone alone significantly reduced edema volume. The combined treatment with edaravone and mild hypothermia reduced both infarct and edema volume. In addition, this treatment provided for the best functional outcome. These results demonstrate that free radical scavenger, edaravone attenuates brain edema and that the combined therapy with edaravone and mild hypothermia significantly reduces not only edema but also infarct on transient focal cerebral ischemia in rats. The neuroprotective effects seen in this study may be due to the combined interaction of antiedema activity between edaravone and mild hypothermia, suppressing free radical production. PMID- 14753436 TI - Therapeutical efficacy of a novel non-peptide bradykinin B2 receptor antagonist on brain edema formation and ischemic tissue damage in focal cerebral ischemia. AB - OBJECTIVE: Bradykinin has been identified as a mediator of secondary brain damage in acute insults. We currently studied neuroprotective properties of a bradykinin B2 receptor antagonist (LF16-0687 Ms) in transitory focal cerebral ischemia to assess infarct formation and the development of brain edema. MATERIAL AND METHODS: 55 Rats were subjected to 90 min of MCA-occlusion. The receptor antagonist was administered at two dose levels, given from 30 min prior to ischemia over two days after ischemia. Ischemic tissue damage was quantified at day 7 after MCA-occlusion together with assessment of brain edema in separate experiments. Neurological recovery was studied daily. RESULTS: Animals receiving treatment (low dose) had a better functional recovery, particularly at days 3 and 4 (P < 0.05). Infarct formation was significantly attenuated in these animals in both total and cortical brain tissue by 50, or 80%, respectively. Postischemic brain swelling was significantly lowered, i.e. by 62%. CONCLUSIONS: Our findings provide further support for a mediator role of bradykinin in ischemic brain damage including edema formation, obviously by ligand binding to the bradykinin B2 receptor. The availability of a receptor antagonist may afford opportunity for translation of this experimental treatment into stroke patients. PMID- 14753437 TI - Iodoamphetamine (IMP) uptake in the brain is increased after experimental cerebral venous hypertension in the rat. AB - Cerebral venous hypertension (VH) is a pathological condition associated with arteriovenous malformation, and is an important determinant factor for clinical symptom and outcome. Using a rat whole brain VH model created by a carotid jugular fistula and contralateral jugular vein ligation we measured superior sagittal sinus pressure (SSSP) and cerebral blood flow (CBF) by the 14C iodoamphetamine (IMP) indicator fractionation technique under isoflurane anesthesia with controlled ventilation. SSSP was significantly elevated in the fistula group (15 +/- 1 mmHg) (mean +/- SEM) compared to the vessel occlusion (5 +/- 1) and sham operated groups (5 +/- 1) (P < 0.05). Mean absolute CBF in the fistula (103 +/- 3 ml/ min/100 g) and vessel occlusion (90 +/- 4) groups were elevated compared to the sham group (77 +/- 3) (P < 0.05). Physiological parameters (mean blood pressure, heart rate, and blood gas analysis) before measurement were not different. In this VH model we found increases in SSSP and IMP uptake in the brain. We assume the mechanism of IMP uptake increase in VH animal may be slow circulation with a possible change of first pass extraction of IMP. PMID- 14753438 TI - Vasogenic edema and VEGF expression in a rat two-vein occlusion model. AB - Vasogenic edema plays an important etiologic role in the pathogenesis of cerebral venous circulation disturbances (CVCDs). Since vascular endothelial growth factor (VEGF) is a major mediator in angiogenesis and vascular permeability, including induction of vasogenic edema, the present study was undertaken to investigate whether it has any relevance to CVCDs. Male Wistar rats (n = 15) were used. Two adjacent cortical veins were occluded photochemically using rose bengal dye and fiberoptic illumination, with evaluation 24 hours thereafter by magnetic resonance imaging (MRI). Each brain was removed from the skull immediately after MRI and processed for hematoxylin-eosin staining (H&E staining) of sections for histopathology and comparison with MRI data. VEGF expression as demonstrated immunohistochemically appeared to coincide with vasogenic edema, diagnosed as high intensity areas on apparent diffusion coefficient of water (ADCw) maps. On the basis of these data, we conclude that VEGF is related to formation of vasogenic edema in the acute stage of CVCD. PMID- 14753439 TI - Astroglial swelling in the neuronal depolarization ensemble. AB - The coupling between neuronal depolarization and astroglial swelling was examined. First, previous in vitro data for the swelling of cultured neurons (N18; rat neuroblastoma) and astroglia (C6; rat astroglyoma) upon exposure to a hypoosmotic solution were reappraised. Neurons swelled rapidly, forming blisters, and easily burst. whereas astroglia resisted swelling and slowly assumed a large full-moon shape. The time constant of swelling was 35.2 +/- 7.8 s for N18 and 594.8 +/- 554.0 s for C6. The glial plasmic membrane was found to be much stronger than the neuronal one, presumably due to a well-developed cytoskeleton. To overcome such neuronal membranous weakness, strong astroglial processes need to cover the neurons including the cell body and synapses, as demonstrated electron-microscopically. Next, in situ astroglial swelling was investigated in rats. During K(+)-induced cortical spreading depression, increases or decreases of a wave-ring spread of light (550 nm) transmission through a 1 mm-thick cerebral cortical layer was observed. The moving local optical density decrease in the cortex was attributable to local vascular bed compression induced by astroglial swelling, since concomitant occurrence of colocated dynamic capillary flow stall was confirmed by a hemodilution technique. Astroglial swelling may occur in an ensemble acting during neuronal depolarization, suggesting that neurons and astroglia behave like a unit complex. PMID- 14753440 TI - An Na+/H+ exchanger inhibitor suppresses cellular swelling and neuronal death induced by glutamate in cultured cortical neurons. AB - We examined the effects of a selective Na+/H+ exchanger inhibitor, SM-20220 (N (aminoiminomethyl)-1-methyl-1H-indole-2-carboxamide methanesulfonate), on neuronal death induced by glutamate in rat cortical neurons. Morphological changes in neurons were observed with a differential interference contrast microscope, and cellular swelling was analysed. Neuronal death was assessed by staining the cell with propidium iodide. The intracellular calcium concentration ([Ca2+]i) and the intracellular pH were measured by fluorescence imaging with fluo-3/AM as an indicator for [Ca2+]i and BCECF/AM for pH, respectively. SM-20220 (0.3 to 30 nM) dose-dependently attenuated glutamate (300 microM)-induced neuronal death in a dose-dependent fashion over 6 hours, and inhibited acute cellular swelling following glutamate (100 microM) exposure. SM-20220 suppressed the persistent [Ca2+]i increase following glutamate (500 microM) exposure, and inhibited intracellular acidification induced by glutamate (1 mM). The activation of the Na+/H+ exchanger system may enhance the progress of cerebral damage and oedema following glutamate exposure. SM-20220, a Na+/H+ exchanger inhibitor, suppressed neuronal death and cellular swelling induced by glutamate through inhibition of both Ca2+ influx and acidification in neurons. PMID- 14753441 TI - Akt phosphorylation and cell survival after hypoxia-induced cytochrome c release in superfused respiring neonatal rat cerebrocortical slices. AB - Phosphorylation of Akt before hypoxia (30 min) and during reoxygenation (4 h) was evaluated in superperfused neonatal rat cerebrocortical slices (350 microm, P7, Sprague-Dawley). Cytosolic cytochrome c intensities in Western blots, which were increased at the end of hypoxia. were decreased during reoxygenation. Western blot intensities of phosphorylated Akt (phospho-Akt), nearly undetectable at the end of hypoxia, recovered quickly during reoxygenation, in a trend opposite that for cytochrome c. At 1.5 h and 4 h after hypoxia they became larger or the same as before hypoxia. Total Akt was unchanged by hypoxia and reoxygenation. Phosphocreatine (PCr) and nucleotide triphosphates (NTP) were measured in parallel 14.1 Tesla ex vivo 31P NMR superfused brain slice studies. PCr and alpha NTP were nearly undetectable at the end of hypoxia. Although they recovered quickly after hypoxia, they were lower than before hypoxia. Reductions in phospho Akt during hypoxia were consistent with the general unavailability of basic high energy phosphates. Preferential phosphorylation of Akt after hypoxia suggested that substantial reductions in intracellular energy, as indicated by PCr and NTP, might be tolerated by processes important for generating phospho-Akt. Additionally, the post-hypoxia increase in phospho-Akt might have contributed to concomitant reductions in cytosolic cytochrome c. PMID- 14753442 TI - The spinal GDNF level is increased after transient spinal cord ischemia in the rat. AB - Glial cell line-derived neurotrophic factor (GDNF) is known as the most potent neurotrophic factor against injury. We have characterized spinal GDNF changes after ischemia to clarify its possible physiological role against ischemic damage. Spinal ischemia in the rat was produced by cross-clamping of the thoracic aorta together with systemic hypotension. The spinal tissue GDNF level was measured by enzyme-linked immunoabsorbent assay (ELISA) and the localization of GDNF in the tissue was examined by immunohistochemistry. GDNF was increased reaching two peaks after ischemia. The first peak was at 2 hrs after onset of recirculation derived from alpha motor neurons. The second GDNF peak was at 72 hrs provided by astrocytes. These data suggest a necessity of GDNF to increase to protect against ischemic damage, and that activated astrocytes may have an important role in maintaining the GDNF level. PMID- 14753443 TI - Identification of malignant brain edema after hemispheric stroke by PET-imaging and microdialysis. AB - Cerebral blood flow (CBF) and extent of irreversible tissue damage as well as the time course of extracellular concentration of amino acids, substrates of energy metabolism, and purine metabolites, intracranial pressure and tissue oxygen tension were assessed in 34 patients with large strokes covering more than 50% of the MCA territory. The results were compared to findings in the experimental model of transient (for 3 hours) MCA occlusion in cats. In the experimental model as well as in the clinical setting development of malignant brain infarcts (due to formation of space occupying brain edema) was predicted by the size of critically hypoperfused tissue and the volume of irreversibly damaged tissue. The course of malignant infarcts was characterized by progressive increase in concentrations of excitatory amino acids, lactate, pyruvate, glycerol, hypoxanthine and in intracranial pressure, while cerebral perfusion pressure and tissue oxygen tension decreased. These results clearly differentiate a malignant from a benign course of large hemispheric infarction. The methods can be used to identify patients at risk for formation of space occupying edema and to select patients who could benefit from invasive therapeutic strategies. PMID- 14753444 TI - Cerebral blood flow index image as a simple indicator for the fate of acute ischemic lesion. AB - BACKGROUND AND PURPOSE: To evaluate the feasibility of utilizing cerebral blood flow (CBF) index images, we attempted to investigate 1) whether CBF index images can reveal the resulting infracted area, 2) whether the CBF index can correlate other modality (SPECT). METHODS: DWI and DPI were obtained in 17 patients within 12 hours of stroke onset and follow up MRI. On three DPI delivered images, namely relative regional cerebral blood volume (rrCBV), uncorrected mean transit time (MTTu) and CBF index images, correlations between initial lesion volume of and follow up infarction volume of three images and rCBF images delivered with singular value decomposition (SVD) methods were assessed. Then 99mTc-ECD SPECT was taken immediately after MRI to correlate to MRI data. RESULTS: Among the three images, lesion volume of CBF index images against follow up infarct volume had the highest correlation (r = 0.995) to a linear fit and the slope was closest to 1.0 (0.91) and had identical accuracy to the regression coefficient of rCBF images. CBF index well correlated to SPECT delivered CBF. CONCLUSION: CBF index images can accurately predict final infarct volume. Evaluating CBF index images together with DWI can guide the initial assessment in the acute stage of cerebral ischemia. PMID- 14753445 TI - Transient elevation of serum bilirubin (a heme oxygenase-1 metabolite) level in hemorrhagic stroke: bilirubin is a marker of oxidant stress. AB - Bilirubin (Bil) is the end product of heme catabolism. The production of Bil reflects heme oxygenase-1 expression in response to oxidative stress in various diseases. To assess the role of Bil as a marker of oxidative stress in cases of brain damage, we measured serum Bil concentrations in patients with hemorrhagic stroke. Serum levels of total Bil were measured in 20 subarachnoid hemorrhage patients with symptomatic vasospasms and in 23 patients with intracerebral hemorrhage; concentrations were measured every day for 14 consecutive days. Serum Bil levels were significantly elevated in the early phases in both groups. Moreover, transient elevation was observed on the day prior to the observation of clinical manifestations of symptomatic vasospasm after SAH. Bil, known to be a powerful antioxidant, was induced after hemorrhagic stroke, reflecting the intensity of oxidative stress. Plasma Bil concentrations might serve as a useful marker of oxidative stress in hemorrhagic stroke patients. PMID- 14753446 TI - Relation between brain oxygen metabolism and temperature gradient between brain and bladder. AB - Brain temperature is slightly higher than systemic core temperature normally. Fluctuations of a temperature gradient between brain and core body have recently been reported after a severe brain insult. The pathophysiological significance of the gradient fluctuations is unclear. This study aims to identify the gradient fluctuations between brain and core temperatures after a brain insult. Temperature gradient (brain temperature minus bladder temperature: degrees C) was measured in 11 patients (125 points) with severe brain injury (4 patients with subarachnoid hemorrhage, 4 with cerebral hemorrhage. and 3 with traumatic brain injury). Cerebral perfusion pressure (CPP; mmHg) and jugular venous blood saturation (SjO2; %) was also measured. The average gradient was 0.29 +/- 0.285 degrees C when CPP was above 50 mmHg. SjO2 was inversely related to the temperature gradient in a significant manner (r = 0.472; P < 0.0001). Temperature gradients tended to increase and then decrease when CPP < 50 mmHg. This study demonstrates that increased temperature gradient has a significant inverse correlation with SjO2 at CPP > 50 mmHg. The current results suggest that the fluctuations in temperature gradient in critical conditions reflect brain ischemia. PMID- 14753447 TI - Neuropeptide release influences brain edema formation after diffuse traumatic brain injury. AB - The mechanisms associated with edema formation after traumatic brain injury (TBI) have not been fully elucidated. In peripheral tissue injury, the neurogenic component of inflammation plays a significant role in increased vascular permeability and edema formation. However, few studies have examined the role of neuropeptide induced neurogenic inflammation following TBI. Adult male Sprague Dawley rats were either left untreated, or pre-treated with capsaicin (125 mg/kg s.c.) or equal volume vehicle, and injured 14 days later using the 2-meter impact acceleration model. Subgroups of animals were assessed for blood brain barrier (BBB) permeability (Evans Blue), brain edema (wet weight/dry weight) and functional outcome (Barnes maze and Rotarod) for up to 2 weeks post-trauma. Increased BBB permeability was present in untreated animals between 3 and 6 h after injury but not at later time-points. Edema was maximal at 5 h after trauma, declined and then significantly increased over the 5 days post-trauma. In contrast, capsaicin pre-treated, neuropeptide-depleted animals exhibited no significant increase in BBB permeability or edema compared to vehicle treated animals after injury. Notably, motor and cognitive impairments were significantly reduced in the capsaicin-pretreated animals. We conclude that neurogenic inflammation contributes to the development of edema and posttraumatic deficits after diffuse TBI. PMID- 14753448 TI - Modulation of aquaporin-4 water transport in a model of TBI. AB - Our Laboratory has pursued the hypothesis that traumatic brain edema is predominantly cellular and recent supportive evidence has been obtained indicating a non-extracellular route for sodium and water entering brain. The aim of this study was to investigate if astrocytic endfeet are involved in this passage, using a potent activator of Protein Kinase C (phorbol ester) to modify and closing the Aquaporin 4 (AQP4), a water channel specific for astrocytic endfoot. Anaesthetized Sprague-Dawley rats were subjected to an intracerebroventricular bolus of phorbol ester (50 pmol/4 microl) or vehicle, in the right hemisphere and after 30 minutes they were exposed to the well established conical contusion model (3 mm depth at 6 m/sec) on the same side. After trauma, they were subjected to 5 hours of drug continuous infusion, then sacrificed. Water content measurements for both right (injured) and left (uninjured) hemispheres were calculated using the wet weight/dry weight technique. Results of these experiments showed a significant decrease in water content in injured phorbol treated animals, underlying that AQP4 regulation plays an important role in brain edema following stroke, and supporting the concept of cellular formation for edema via astrocytic foot processes. PMID- 14753449 TI - Correlation of lesion volume and brain swelling from a focal brain trauma. AB - Brain edema and secondary growth of a traumatic brain tissue necrosis are important manifestations of secondary brain damage and of prognostic significance in severe head injury. Aim of the current study was to analyze the interdependency of the resulting brain swelling from the size of the focal traumatic lesion. Male Sprague-Dawley rats were intubated and mechanically ventilated. A trephination was made over the left parietal cortex for induction of a cold lesion. Different injury severities were achieved by varying the contact time of the cooled copper-cylinder and the exposed cortex. Animals were randomized into 12 experimental groups. Hemispheric brain swelling was measured in groups A1-A6 (n = 4-8) by gravimetry 24 hrs after lesions of six increasing severity levels. Correspondingly, in animals of groups B1-B6 (n = 5-7) the volume of necrosis was planimetrically assessed in histological serial sections of the brain obtained 24 hrs after trauma of different severity. In groups A1-A6. hemispheric brain swelling (increase in weight) was growing with increasing contact duration of the cold probe with the exposed cerebral cortex, i.e. from 7.7 +/- 0.4% (5 s) to a maximum of 9.9 +/- 0.5% (25 s). Longer contact periodes (30 s) were not further effective to increase hemispheric brain swelling. The contact times and extent of swelling were linearly correlated between 5 s and 25 s (r = 0.47; p < 0.01). The volume of necrosis in groups B1-B6 increased from 35.7 +/- 3.7 mm3 (5 s) to 106.3 +/- 10.3 mm3 (30 s). There was again a linear correlation between the duration of contact of the cold probe (i.e. injury severity) with the brain cortex and the volume of necrosis (r = 0.77; p < 0.01). Accordingly. the lesion volume could be increased in a reproducible manner from 35.7 up to 106.3 mm3 by extending the contact times of the cooling device and cerebral cortex. Hemispheric swelling, predominantly due to vasogenic brain edema, was expanding in relationship with the volume of necrosis. PMID- 14753450 TI - A role of glial fibrillary acidic protein in hippocampal degeneration after cerebral trauma or kainate-induced seizure. AB - Astrocytes perform a variety of functions in the adult central nervous system (CNS). Recent evidence suggests the robust upregulation of glial fibrillary acidic protein (GFAP) after CNS insult. However, little is known about the role of GFAP in the hippocampal degeneration after brain injury. We herein compared the GFAP knockout (KO) and wild type (WT) mice on the histological and behavioral outcome in response to cerebral trauma or kainic acid (KA)-induced seizure. Although all KO mice showed hippocampal CA3 neuronal degeneration. WT mice did not show any neuronal degeneration in CA3 subfield at 72 hrs after trauma. Thereafter, KO mice showed a higher susceptibility to KA-induced seizures and an increased number of pyknotic CA3 neurons 72 hrs after KA administration. These results indicate that GFAP plays a crucial role in the hippocampal neurodegeneration after CNS insult. PMID- 14753451 TI - Antioxidant properties of the vasoactive endocannabinoid, 2-arachidonoyl glycerol (2-AG). AB - Reactive oxygen species (ROS) were shown to play a role in altering blood-brain barrier (BBB) permeability and formation of brain edema induced by trauma and/or ischemia. 2-arachidonoyl glycerol (2-AG), a novel, potent vasodilatory and cytoprotective endocannabinoid has been implicated to act as an antioxidative agent. This study examines: 1) the possible 2-AG modulation of BBB injury and edema formation induced by closed head injury (CHI); and 2) comparable effects between 2-AG and 4-hydroxy-2,2,6,6-tetramethylpiperidine-N-oxyl (TPL), a known antioxidant nitroxide on endothelial Ca2+ and cytoskeletal responses to H2O2 (ROS). 2-AG treatment reduced the CHI-induced increase in BBB permeability and brain edema. The endothelial H2O2-stimulated Ca2+ mobilization and cytoskeleton (vimentin) rearrangement was modified by either 2-AG or TPL. These findings provide evidence of 2-AG antioxidant activity and are consistent with the involvement of ROS in the pathomechanism of CHI-induced BBB injury and brain edema. PMID- 14753452 TI - Very early expression of vascular endothelial growth factor in brain oedema tissue associated with brain contusion. AB - BACKGROUND: Brain oedema associated with cerebral contusion can be life threatening. Mechanisms of the development of brain oedema are still unclear. METHOD: We investigated the expression of vascular endothelial growth factor (VEGF) and its receptor VEGFR-2 (KDR/Flk-1) in the contusional brain tissue obtained during neurosurgery from 5 patients. FINDINGS: VEGF is expressed in some but not all the astrocytes, and KDR/Flk-1 is expressed in vascular endothelial cells in the con-tusional tissue as early as 3 hours after onset. CONCLUSION: The results suggested that the VEGF is induced in the contusional tissue in the very early period after onset, and that it increases capillary permeability via KDR/Flk-1 resulting in vasogenic type brain oedema. PMID- 14753453 TI - Changes of perivascular macrophages in the process of brain edema induced by cold injury. AB - Perivascular macrophages are considered as cerebral scavenger cells under physical and pathological conditions. In this study, we tried to examine changes of perivascular macrophages, especially changes of the characteristic lysosomal inclusion bodies that are rich in hydrolytic enzymes, in the process of brain edema induced by cold injury. Wistar male rats aged 4 months were treated with dry ice for 20 minutes through a drilled hole at the parietal bone. At different time points after the cold injury, cerebral cortex was excised and the immunoreaction for cathepsin D, one kind of lysosomal protease, was examined by post-embedding immuno-electron microscopy. The reactions of cathepsin D were located in the inclusion bodies of perivascular macrophages. At 5 and 10 hours after cold injury, the reactions increased dramatically. Then the reactions inclined to decrease, and reached the minimum at 1 week after cold injury. The reactions seemed to recover at 2 weeks after cold injury. The changes of cathepsin D reactions suggest that the function of perivascular macrophages as scavenger cells were activated in the early phase of the process of brain edema, their later declines might be caused by severe pathological conditions. PMID- 14753454 TI - Mitogen-activated protein kinases phosphorylation in posttraumatic selective vulnerability in rats. AB - Mitogenic stimulation of the Mitogen-activated protein kinase (MAPK) pathway modulates the activity of many transcriptional factors leading to biological responses. Of these, three MAPK cascades are well characterized as extracellular signal-regulated protein kinase (ERK), c-Jun NH(2)-terminal kinase (JNK), and p38 pathways. The aim of this study was to investigate the topographic distribution and the role of activated MAPK pathways after fluid percussion injury (FPI) in rats. In the present results, FPI significantly induced ERK- and JNK phosphorylation, but not p38-phosphorylation in the cortex and hippocampus at the injury site. The immunoreactivity for phospho-ERK was localized in the superficial neuronal layers, dentate hilar neurons, and the damaged CA3 neurons after 30 mins of FPI. Double immunostaining showed that phospho-ERK was prominent in astrocytes 6 hrs after TBI. The current results suggest that MAPK pathways are involved in signal transduction after FPI. PMID- 14753455 TI - Treatment of cold injury-induced brain edema with a nonspecific matrix metalloproteinase inhibitor MMI270 in rats. AB - Blood-brain barrier (BBB) disruption is a critical event leading to vasogenic brain edema and secondary brain damage after cold injury-induced brain trauma. Matrix metalloproteinases (MMPs) are implicated in BBB disruption in this model. The purpose of this study was to examine the effects of MMI270, a synthetic nonspecific MMP inhibitor, on cold injury-induced brain edema in rats. Treatment with MMI270, a bolus injection at a dose of 30 mg/kg, was started immediately after the induction of cold injury and was continued at a dose of 40 mg/kg/day using an intraperitoneal osmotic minipump. At 24 hours after the cold injury, the brain water content and the BBB permeability to Evans Blue (EB) were determined. The secondary brain lesion was assessed using hematoxylin and eosin (H-E) staining at 7 days after the cold injury. Compared with the untreated control group, treatment with MMI270 significantly reduced the brain water content in the ipsilateral core and intermediate areas and protected the BBB integrity to EB in the ipsilateral core area. The secondary lesion was significantly smaller in the MMI270-treated animals compared with the untreated animals. Our results indicate that treatment with MMI270 in rats exhibits protection in acute brain edema formation and secondary brain lesion by attenuating the BBB permeability after cold injury. PMID- 14753456 TI - Prions prevent brain damage after experimental brain injury: a preliminary report. AB - The physiological function of the normal cellular form of prion protein (PrPC) is not yet fully understood. In the current study we used prion protein gene knock out mice (Prnp-/-) to assess the role of PrPC in traumatic brain injury. Prnp+/- and Prnp-/- mice were subjected to weight-drop contusional brain injury over the left parietal cortex. Prnp-/- mice manifested a significantly larger lesion volume and worse neuromotor scores than did their Prnp+/- littermates. IgG immunostaining revealed that in Prnp-/- mice the breakdown in the blood-brain barrier (BBB) was more extensive at 1 month after brain injury. Our results are in agreement with previous in vitro findings of the neuroprotective role of PrPC and further support the hypothesis that functional loss of PrPC plays a pathogenic role in prion diseases. We also suggest that PrPC modulates BBB function. PMID- 14753457 TI - The effect of cyclosporin A on brain edema formation following experimental cortical contusion. AB - Cyclosporin A (CsA) has been shown by our laboratory and others to be neuroprotective in the experimental animal model of traumatic brain injury. However, we found that the intrathecal administration resulted in a concomitant increase of brain edema. The aim of this study was to assess whether intravascular administration may also influence brain edema formation. This project includes two independent series in which different doses of CsA were intravenously given to Sprague-Dawley rats of each group. In the first series, the animals were exposed to focal brain injury by a controlled cortical impact (CCI, 6 m/sec, 3 mm depth) and randomized into the following two groups: 20 mg/kg CsA and control vehicle. In the second series, animals were also injured by CCI and randomized into 35 mg/kg CsA and control vehicle. The intravenous continuous (1 h) infusion was begun 30 minutes after the insult. All animals were sacrificed at 24 hours post injury to assess the brain water content using the gravimetric method. Intravenously-administrated CsA of either 20 or 35 mg/kg did not significantly change the brain water content. We therefore suggest that an intravascular route may be better for CsA administration because the intrathecal injection may exacerbate brain edema as found in our previous study. PMID- 14753458 TI - The glutamate AMPA receptor antagonist, YM872, attenuates regional cerebral edema and IgG immunoreactivity following experimental brain injury in rats. AB - We previously reported the neuroprotective effects of the glutamate AMPA receptor antagonist YM872 on neurobehavioral motor function and cortical tissue loss (lesion volume) in a brain-injured rat model. Here we examined its effect on brain edema and the breakdown of the blood-brain barrier (BBB). Rats subjected to severe right lateral (parasagittal) fluid-percussion brain injury or sham injury received a 4-hr intravenous infusion of YM872 (20 mg/kg/ hr, 20 mg/3 ml) or normal saline starting at 15 min post-injury. At 48 hr we removed their brains and evaluated the cerebral regional edema by the wet weight/dry weight method. Another group of rats was transcardially fixed with 10% formalin at 2 weeks after injury. Serial brain sections were immunostained for endogenous IgG and the extent and intensity of staining were evaluated. The administration of YM872 resulted in a significant reduction in regional cerebral edema in the injured parietal cortex and a markedly reduced area of IgG immunoreactivy in the injured cortex. Our results indicate that the post-traumatic administration of YM872 may be neuroprotective by reducing BBB breakdown and regional cerebral edema. PMID- 14753459 TI - Effects of edaravone on experimental brain injury in view of free radical reaction. AB - The purpose of this study is to clarify the influence of a novel free radical scavenger edaravone on experimental brain injury. Male Wistar rats were anesthetized with 1-2% halothane. Brain injury was produced using a controlled cortical impact injury device. Experimental rats were divided into 2 groups. In the edaravone group, edaravone (3 mg/kg) was twice administered intravenously for 30 minutes. In the saline group, saline solution was administered in the same way. This administration of edaravone or saline solution made it possible to evaluate the relative effects of edaravone by assessment of free radical reaction and water content. As a result, the level of oxygen free radicals at 50 minutes after brain injury was significantly lower in the edaravone group than in the saline group. The water content in the injured brain at 180 minutes was significantly lower in the edaravone group than in the saline group. Therefore, we propose that edaravone may be effective for treatment in head injury. PMID- 14753460 TI - Antioxidant compounds EGB-761 and BN-52021 attenuate brain edema formation and hemeoxygenase expression following hyperthermic brain injury in the rat. AB - Role of carbon monoxide (CO) in hyperthermic brain injury induced brain pathology was examined in a rat model using immunohistochemistry of the hemeoxygenase-2 (HO 2) enzyme. Exposure of rats to 4 h heat stress at 38 degrees C resulted in profound hyperthermia, breakdown of the blood-brain barrier (BBB), brain edema formation, cell damage and expression of HO-2 in several brain regions. Pretreatment with potent antioxidant compounds EGB-761 and BN-52021 markedly reduced the HO-2 expression, BBB breakdown, brain edema formation and cell damage without attenuating the hyperthermic response. This effect was most marked in animals treated with EGB-761. These observations suggest that upregulation of HO 2 representing generation of CO plays important roles in hyperthermic brain injury, and oxidative stress seems to be one of the most important signals in inducing HO-2 expression in hyperthermia, not reported earlier. PMID- 14753461 TI - Edema fluid accumulation within necrotic brain tissue as a cause of the mass effect of cerebral contusion in head trauma patients. AB - The early massive edema caused by severe cerebral contusion results in progressive intracranial pressure (ICP) elevation and clinical deterioration within 24-72 hours post-trauma. Surgical excision of the necrotic brain tissue represents the only therapy, which can provide satisfactory control of the elevated ICP and clinical deterioration. In order to elucidate the mechanisms underlying the early massive edema, we have carried out a series of detailed clinical studies. Diffusion magnetic resonance (MR) imaging and apparent diffusion co-efficient (ADC) mapping suggest that cells in the central area of contusion undergo shrinkage, disintegration and homogenization, whereas cellular swelling is predominant in the peripheral area during the period of 24-72 hours post-trauma. The ADC values in the central and peripheral areas are maximally dissociated during this period. A large amount of edema fluid accumulates within the necrotic brain tissue of the central area beginning at approximately 24 hours post-trauma. We have found that fluid-blood interface formation within the central area does not represent an uncommon finding in various neuroimaging examinations of cerebral contusions, indicating layering of red blood cells within the necrotic brain tissue accumulating voluminous edema fluid. Intravenous slow infusion of gadolinium-DTPA and delayed MR imaging revealed that the central area of contusion can be enhanced at 24-48 hours post-trauma. implying that water supply from the blood vessels is not completely interrupted. Necrotic brain tissue sampled from the central area of contusion during surgery demonstrates a very high osmolality. It appears that the capacitance for edema fluid accumulation increases in the central area, whereas cellular swelling in the peripheral area elevates the resistance for edema fluid propagation. Combination of these circumstances may facilitate edema fluid accumulation in the central area. We also suggest that the dissociation of ADC values and high osmolality within the necrotic brain tissue may generate an osmotic potential across the central and peripheral areas and contribute to the early massive edema caused by cerebral contusion. PMID- 14753462 TI - Ultra-early study of edema formation in cerebral contusion using diffusion MRI and ADC mapping. AB - OBJECTIVE: Our previous studies have reported that heterogeneous mechanisms exist in early edema formation in cerebral contusion, and cytotoxic edema plays an important role within 48 hours post-trauma. It is remains unclear, when edema begins to develop following injury. In order to determine the time course of edema development, diffusion imaging and ADC (apparent diffusion co-efficient) mapping was performed in 10 patients within 24 hours post-trauma with cerebral contusion. METHODS: Diffusion imaging and ADC mapping were performed employing 1.0 T echo planar MRI. ADC values were indicated as a ration relative to the values of intact brain areas. RESULTS: Within 3 hours post-trauma, diffusion MRI showed no remarkable changes, and the ADC values were within normal limit (ADC ratio (=contused/normal brain) = 1.00 +/- 0.21, (mean +/- SD)). At 6 hours post trauma, diffusion images demonstrated a low intensity core in the contusion proper and a high intensity rim in the peripheral area of contusion. The ADC value increased in the contusion proper (ADC ratio = 1.26 +/- 0.13) and decreased in the peripheral area (ADC ratio = 0.58 +/- 0.19). CONCLUSIONS: These findings indicated that early cellular swelling in the peripheral area of contusion begins within 6 hours following injury. This delayed occurrence of contusion-induced cellular swelling suggests that the CBF does not decrease to ischemic level immediately following injury. PMID- 14753463 TI - Mixed dishomogeneous hemorrhagic brain contusions. Mapping of cerebral blood flow. AB - The aim of the study was to verify whether regional cerebral blood flow (rCBF) was distributed centrifugally in traumatic hemorrhagic contusions with multiple cores within an oedematous area. Seventeen traumatic brain contusions, from 14 patients with severe head injury (GCS < 9), were analyzed during 39 Xenon enhanced computerized tomography (Xe-CT) studies. The CBF was measured in 3 concentric regions of interest (ROls): the hemorrhagic core, the intracontusional oedematous low density area and a 1 cm rim of pericontusional normal-appearing brain tissue surrounding the contusion. Differences between rCBFs in the three ROIs were found (p < 0.0001). rCBF in both the hemorrhagic core (21.4 +/- 19.4 ml/ 100gr/min) and the intracontusional low density area (28.4 +/- 19 ml/100gr/min) were lower than rCBF in pericontusional normal-appearing area (41.9 +/- 16 ml/100gr/min) (p < 0.0001). No significant differences were found between rCBF measured in the hemorrhagic core and intracontusional low density area (p = 0.184). Our study suggests that in the mixed density contusions with multiple hemorrhagic cores, the CBF is concentrically distributed, improving from the core to the periphery. PMID- 14753464 TI - Neuronal damage in pericontusional edema zone. AB - In this study, we investigated the molecular biological and histopathological aspects of the etiological mechanisms for pericontusional edema zone (PEZ). The subjects were 5 patients with traumatic brain injury who underwent surgery to evacuate the resulting hematoma. The average age of the subjects was 52 +/- 27.5 years. The GCS at the time of admission was 5-9. At operation apart from evacuating the hematoma, the PEZ was also excised and then examined histopathologically. Cerebrospinal fluid (CSF) levels of IL-6, IL-8, and IL-10 were measured at the time of admission and at 24 and 72 hours. Histological examination revealed large numbers of neutrophils accumulating within blood vessels in the PEZ, with some focal migration. IL-6: CSF levels at the time of admission and at 24, 72, and 72 hours were 550, 4350, and 878000 pg/ml, respectively (median values). IL-8: CSF levels were 715, 804, and 24900 pg/ml, respectively. IL-10: CSF levels were 15, 4, and 5 pg/ml, respectively. High levels of IL-6 and IL-8 were seen from an early stage, and became markedly higher with enlargement of the PEZ. The PEZ is thought to be due to microvascular disturbance by neutrophils stimulated by inflammatory cytokines, and neuronal damage from migrated neutrophils. PMID- 14753465 TI - Characteristics of parietal-parasagittal hemorrhage after mild or moderate traumatic brain injury. AB - The aim of this study is to clarify the clinical characteristics of parietal parasagittal traumatic intracranial hemorrhage (TICH) after mild or moderate traumatic brain injury (TBI). METHODS; Subjects were 105 patients with mild or moderate TBI. The patients with parietal-parasagittal TICH were clinically analyzed based on the initial brain CT findings, hematoma sites and the clinical course as compared to those with TICH at other sites. RESULTS: Hematoma was located in the frontal or temporal lobes in 89.5% of the subjects and the parietal-parasagittal in 10.5%. Ten of the 11 patients suffering parietal parasagittal TICH had skull fractures (7 depressed and 3 linear) but no depressed fracture observed in patients with frontal or temporal lobe hemorrhage. Neurological deterioration leading to a comatose state more frequently occurred in 63.6% of patients with parietal-parasagittal TICH than in those with frontal or temporal lobe hemorrhage (19.1%, p < 0.01). The incidence of hematoma growth was significantly higher in patients with parietal-parasagittal TICH (63.6%) than in those with frontal or temporal lobe hemorrhage (31.9%, p < 0.05). DISCUSSION: The incidence of parietal-parasagittal TICH is low, but the risk of neurological deterioration due to hematoma enlargement is significantly high. Parietal parasagittal TICH may differ clinically from frontal-temporal TICH. PMID- 14753466 TI - IL-8 is a key mediator of neuroinflammation in severe traumatic brain injuries. AB - The subjects were 22 patients with severe head injury. The average age was 45 +/- 18.3 years. There were 13 survivors and 9 fatalities. Samples of peripheral blood and cerebrospinal fluid (CSF) were taken four times, at the time of admission and at 24, 72, and 168 hours later. IL-6: For the survivor group, peripheral blood levels were 181, 105, 37, and 26 pg/ml, respectively (median values). CSF levels were 5376, 3565, 328, and 764 pg/ml, respectively. For the fatality group, peripheral blood levels were 102, 176, 873, and 3059 pg/ml, respectively, whereas CSF levels were 15241, 97384, 548225, and 366500 pg/ml, respectively. IL-8: For the survivor group, peripheral blood levels were 36, 15, 15, and 15 pg/ml, respectively, whereas CSF levels were 23736, 4074, 355, and 1509 pg/ml, respectively. For the fatality group, peripheral blood levels were 21, 28, 43, and 77 pg/mL, respectively, whereas CSF levels were 29003, 8906, 5852, and 8220 pg/ml, respectively. IL-6 and IL-8 levels were significantly higher after 72 hours in the fatality group. The fact that CSF IL-8 was 1000 times that in the peripheral blood at the time of admission, and decreased thereafter, indicates that IL-8 is a key mediator of neuroinflammation. PMID- 14753467 TI - The risk factors for the occurrence of acute brain swelling in acute subdural hematoma. AB - There are many cases of acute subdural hematoma (ASDH) in which acute brain swelling (ABS) occurs intra- or post-operative period and the control of intracranial pressure becomes impossible, resulting in death. Consequently, we investigated the risk factors for ABS in ASDH. Thirteen cases were used for this study. The average age was 53.8 years, and all the subjects were males whose Glasgow Coma Scale (GCS) was an average of 4.8 when hospitalized. We examined the relationship of blood gas values (pH, PaO2, PaCO2, HCO3-, and base excess) when hospitalized, mean arterial blood pressure (MAP) before surgery, and the presence of hypotension during the surgery with ABS. There were no cases, which showed evidence of hypoxia or hypotension when hospitalized or before the surgery. No statistical differences in blood gas values when hospitalized were observed between the 7 cases with ABS and the 6 cases without ABS. However, all of 7 cases with ABS showed MAP > or = 140 mmHg, and 6 cases demonstrated intraoperative hypotension (systolic pressure < 90 mmHg). Conspicuous hypertension before the surgery was shown to be a possible risk factor for ABS. Therefore; MAP > or = 140 mmHg may be a warning sign for the occurrence of ABS. PMID- 14753468 TI - Volume-targeted therapy of increased intracranial pressure. AB - Fluid exchange across the intact blood-brain barrier (BBB) is counteracted by the low permeability to crystalloids (mainly Na+ and Cl-) combined with the high osmotic pressure (5,700 mm Hg) on both sides of the BBB. If the BBB is disrupted transcapillary water transport will be determined by the differences in hydrostatic and colloid osmotic pressure between the intra- and extracapillary compartments. Under these pathological conditions pressure autoregulation of cerebral blood flow is likely to be impaired and intracapillary hydrostatic pressure will depend on variations in systemic blood pressure. The volume targeted "Lund concept" can be summarized under four headings: A. Reduction of stress response and cerebral energy metabolism: B. Reduction of capillary hydrostatic pressure; C. Maintenance of colloid osmotic pressure and control of fluid balance: D. Reduction of cerebral blood volume. The efficacy of the protocol has been evaluated in experimental and clinical studies regarding the physiological and biochemical (utilizing intracerebral microdialysis) effects and the clinical experiences have been favourable. PMID- 14753469 TI - Induced acute arterial hypertension and regional cerebral flow in intracontusional low density area. AB - Traumatic brain contusions have been associated with regional ischemia. We aimed to measure the effect of induced supra-normal values of cerebral perfusion pressure (CPP) on regional cerebral blood flow (rCBF) in the intracontusional low density area surrounding the contusional hemorrhagic core. In 7 severely head injured patients (GCS < or = 8) harbouring a contusion larger than 2 cm, the rCBF levels were measured, by means of Xenon-enhanced CT, in: 1) the intracontusional low density area: 2) contralaterally, in a normal brain symmetric area. CBF studies were performed at a baseline CPP of 65.3 mmHg +/- 7 and after 20 minutes of norepinephrine-induced CPP supernormal values (88.3 mmHg +/- 10.5) (p = 0.0013). A "paradoxical" reduction of rCBF levels was observed in both the intracontusional low density area (p = 0.07) and the contralateral "normal" area (p = 0.08). In particular, this decrease of rCBF in the intracontusional low density area (-25.7 + 10 ml/100gr/min) (p = 0.0009) was present in only 4 cases, having a mean rCBF at baseline of 25 +/- 16 ml/100gr/min. In the remaining 3 cases in which rCBF at baseline was abnormally low (12 +/- 7 ml/ 100gr/min), rCBF values improved slightly (3.6 +/- 2 ml/100gr/min) (p = 0.61). An acute increase of CPP seems to marginally affect rCBF in the intracontusional low density area having critically reduced initial values, but may greatly reduce rCBF in subjects starting from non-critical baseline values. PMID- 14753470 TI - The importance of decompressive craniectomy for the management of severe head injuries. AB - Neurosurgical therapy aims to minimize the secondary brain damage after a severe head injury. This includes the evacuation of an intracranial space occupying bleeding, the reduction of intracranial volumes, in hematocephalus an external ventricular drainage, and the conservative therapy in order to influence an increased intracranial pressure (ICP) and a decreased p(ti)02. When conservative treatment fails to act a decompressive craniectomy might be successful in lowering ICP. From September 1997 until August 2001 we operated on 439 patients with severe head injuries. 50 patients (11%) were treated by means of a decompressive craniectomy. The prognosis after decompression depends on the clinical signs and symptoms on admission, the patients age and the existence of major extracranial injuries. Our guidelines for an indication for decompressive craniectomy after failure of conservative interventions and evacuation of space occupying hematomas include a patients age below 50 years without multiple trauma, a patients age below 30 years in the presence of major extracranial injuries, a severe brain swelling on CT scan, the exclusion of a primary brainstem lesion or injury and the intervention before irreversible brainstem damage and secondarily while monitoring ICP and p(ti)02 in an interval up to 48 hours after the accident before irreversible brainstem damage or generalized brain damage has occurred. PMID- 14753471 TI - Importance of hemodynamics management in patients with severe head injury and during hypothermia. AB - OBJECTIVE: To evaluate the hemodynamics in patients with traumatic brain injury (TBI) during therapeutic hypothermia. METHODS: Subjects were 25 patients with TBI (GCS; 8 or less). Mean arterial blood pressure (MAP), cerebral perfusion pressure (CPP), cardiac index (CI), systemic oxygen delivery (DO2), systemic vascular resistance index (SVRI), and pulmonary capillary wedge pressure (PCWP) were measured. Patients were retrospectively divided into 3 groups: normothermia (n = 5; NT), and survivors (n = 14; HT-S) and non-survivors (n = 6; HT-Non-S) after hypothermia. and hemodynamics were investigated for difference among groups at 24 hours from induction of normothermia or hypothermia. RESULTS: CPP target was above 70 mmHg, however, HT-Non-S could not maintain CPP above 70 mmHg. The low CPP was the result of elevated ICP, low MAP (P < .05), or both during hypothermia. In HT-Non-S, significantly high SVRI and low CI (P < .05) causing dehydration were observed during cooling. DO2 could not be maintained in HT-Non-S during hypothermia. CONCLUSIONS: These results suggest that patients run the risk of impairing hemodynamics during therapeutic hypothermia. Hemodynamic management is essential during hypothermia. If dehydration occurs during hypothermia. MAP may be reduced due to inadequate sedation, analgesia, and excess use of diuretic agents. PMID- 14753472 TI - Changes in cerebrovascular response during brain hypothermia after traumatic brain injury. AB - OBJECTIVE: Transcranial Doppler sonography (TCD) provides a rapid and noninvasive assessment of cerebral hemodynamics after traumatic brain injury. This study evaluates the change of cerebrovascular response at the rewarming stage of brain hypothermia using TCD. MATERIAL AND METHODS: Seventeen patients who had suffered from closed brain injury were investigated with daily TCD recordings and the changes in flow velocities were evaluated to determine whether they reflected the temperature during brain hypothermia. All patients who had treated brain hypothermia underwent continuous monitoring of SjO2, mean arterial blood pressure, and intracranial pressure (ICP). RESULTS: No significant changes in ICP and cerebral perfusion pressure (CPP) were recorded in all cases. Of 17 patients, 5 had a significant increase in SjO2 > 75% or more, retrospectively, with a robust increase in flow velocities of the middle cerebral artery (P < .01) at the rewarming stage. These cases marked a decrease in pulsatility index (P < .05) concomitant with an increase in SjO2 values. The CT scan revealed acute brain swelling in these cases. CONCLUSION: These data suggest cerebral vasoactivity could be altered at the rewarming stage. Hypothermia runs a risk of hyperemia at the rewarming stage, which induced a decrease in cerebral vasoresistance. PMID- 14753473 TI - Neurotrophic factors attenuate microvascular permeability disturbances and axonal injury following trauma to the rat spinal cord. AB - Alterations of the blood-spinal cord barrier (BSCB) following spinal cord injury (SCI) and leakage of serum proteins induce vasogenic edema and cell damage. The possibility that two members of the neurotrophin family, BDNF or IGF-1 induce neuroprotection by attenuating the BSCB permeability following trauma was examined in a rat model. Repeated topical application of BDNF or IGF-1 (0.1 1microg, 0.5 microg or 1 microg in 10 microl) onto the spinal cord 30 min before SCI or 2, 5, 10 or 30 min thereafter significantly attenuated BSCB permeability to Evans blue and iodine. In the neurotrophin treated rats. edema formation, degradation of MBP, and myelin vesiculation were much less frequent compared to the untreated traumatised rats. The protective effect of BDNF and IGF-1 was most pronounced at the high dose (1 microg in 10 microl) given either 30 min before or within 10 min after SCI. The observations suggest that early intervention with neurotrophins in high doses following trauma (within 10 min) attenuates disturbances of the fluid microenvironment of the spinal cord. This indicates that BSCB opening plays an important role in SCI induced myelin vesiculation and cord pathology. PMID- 14753474 TI - Depletion of endogenous serotonin synthesis with p-CPA attenuates upregulation of constitutive isoform of heme oxygenase-2 expression, edema formation and cell injury following a focal trauma to the rat spinal cord. AB - The possibility that the upregulation of hemeoxygenase (HO) enzyme responsible for carbon monoxide (CO) formation in the spinal cord following trauma is involved in edema formation and cell damage was examined in a rat model. A focal trauma to the rat spinal cord by making an incision into the right dorsal horn of the T10-11 segment resulted in profound upregulation of HO-2 (the constitutive isoform of the enzyme) expression in the T9 and T12 segments 5 h after injury. In these segments a marked increase in edema formation, nerve cell damage, and expression of heat shock protein (HSP 72) were observed. Pretreatment with p chlorophenylalanine (p-CPA, a serotonin synthesis inhibitor) significantly attenuated the trauma induced edema formation, cell injury, and HSP expression. Upregulation of HO-2 in p-CPA treated traumatised rats was considerably reduced. These observations suggest that (i) spinal cord injury has the capacity to induce an upregulation of HO-2 and HSP expression, (ii) abnormal production of CO as reflected by HO-2 expression is injurious to the cord, and (iii) that endogenous serotonin is involved in HO-2 expression in the cord. PMID- 14753475 TI - High frequency electrical stimulation attenuates progressive necrosis and cavitation following spinal cord injury. AB - This study evaluates the influence of preconditioning by following electrical stimulation on primary and secondary lesion formation following spinal cord injury in rats. The dorsal surface of the spinal cord was stimulated (500 Hz. 10 pulses/train, inter train interval of 10 sec. for 2 hrs) at the T7 level 24 hrs before a right side hemisection, carried out immediately after injury and maintained every 24 hrs for 7 days. Preconditioning by electrical stimulation of the spinal cord activates reactive astrocytes and significantly attenuates edema and progressive necrosis and cavitation, concerning especially the primary (1, 3 weeks post injury) and secondary (24 hrs, 1, 3 weeks post injury) lesion volume. The results suggest that pre-conditioning by electrical stimulation prevents spinal cord secondary lesion formation after injury, and that the beneficial effect is provided by astroglial cells with regard to their ability to attenuate trauma induced cellular cascades. PMID- 14753476 TI - Low molecular weight compounds with affinity to melanocortin receptors exert neuroprotection in spinal cord injury--an experimental study in the rat. AB - The possibility that five new low molecular weight compounds with varying affinity and selectivity to the melanocortin receptors will exert neuroprotective effects in the spinal cord injury (SCI) induced edema formation and cell damage was examined in a rat model. A focal trauma of the rat spinal cord made by an incision into the right dorsal horn (T10-11) resulted in profound edema formation, leakage of Evans blue albumin and cell injury of the T9 segment at 5 h. Topical application of the Melacure compound ME10501 in high doses (10 microg in 10 microl) given 5 min after SCI resulted in most significant neuroprotection of the T9 segment of the cord compared to other compounds. Thus, marked reduction in water content, leakage of Evans blue albumin, and cell injury were observed in ME10501 treated traumatised rats. These observations suggest that the non-peptide compound ME10501 with affinity to the melanocortin receptor MC4 is capable to induce neuroprotection in the spinal cord following trauma not reported earlier. PMID- 14753477 TI - Topical application of TNF-alpha antiserum attenuates spinal cord trauma induced edema formation, microvascular permeability disturbances and cell injury in the rat. AB - The possibility that antiserum to tumour necrosis factor-alpha (TNF-alpha) is neuroprotective in spinal cord injury (SCI) was examined in a rat model. SCI was produced by making an incision into the right dorsal horn at the T10-11 segments. Top TNF-alpha antiserum at three concentrations (1:10; 1:50 and 1:100) given 30 min before or 2 min, 5 min or 10 min after trauma resulted in marked reduction in visible swelling, edema formation, and leakage of radiolabelled iodine tracer within the T9 and T12 segments at 5 h in a dose dependent manner. This neuroprotective effect was most pronounced when the antiserum at the highest dose level (1:10) was applied 10 min after SCI. The TNF-alpha antiserum also reduced the SCI induced upregulation of neuronal nitric oxide synthase (nNOS) immunoreactivity in a concentration dependent manner. Taken together, these results suggest that local application of TNF-alpha antiserum is neuroprotective in SCI and that this effect is mediated through NOS regulation. PMID- 14753478 TI - A new antioxidant compound H-290151 attenuates spinal cord injury induced expression of constitutive and inducible isoforms of nitric oxide synthase and edema formation in the rat. AB - The role of oxidative stress in spinal cord injury (SCI) induced upregulation of constitutive or inducible isoforms of nitric oxide synthase (cNOS or iNOS) is not well known. The present investigation was undertaken to examine the influence of an antioxidant compound H-290/51 (Astra-Zeneca, Molndal, Sweden) on SCI induced cNOS and iNOS upregulation in a rat model. SCI induced by incision into the right dorsal horn of the T10-11 segment resulted in marked NOS upregulation. Upregulation of cNOS was most prominent in the uninjured T9 and T12 segments. On the other hand, iNOS expression was most marked in the injured T10-11 segments. These NOS immunoreactivities were mainly confined to the injured cells located in the edematous regions of the cord exhibiting profound leakage of Evans blue and [131]Iodine-sodium tracers. Pre-treatment with H-290/51 markedly attenuated the trauma-induced cNOS and iNOS expression along with the microvascular permeability disturbances, edema formation and cell injury. These results suggest that (i) oxidative stress is involved in SCI induced induction of cNOS and iNOS, (ii) NO plays an important role in the cord pathology, and (iii) that the compound H 290/51 has a potential therapeutic value in SCI. PMID- 14753479 TI - Src family kinase inhibitor PP1 improves motor function by reducing edema after spinal cord contusion in rats. AB - Following spinal cord injury vascular permeability increases around the area of injury, which possibly leads to secondary tissue damage. Vascular endothelial growth factor (VEGF) and Src which exists downstream of VEGF may contribute to edema formation. We here report that the Src family kinase inhibitor PPI could reduce edema and the inflammatory response after spinal cord injury. In this study we have examined the effect of PPI on motor function after mild spinal cord compression injury. We utilized a mild spinal cord compression model in rats. PPI or vehicle only was administered intraperitoneally after cord compression. The motor function of the hind limbs after injury was categorized into 7 grades. At 1, 3, 7 and 14 days after injury, the spinal cord was removed and the extent of edema formation and inflammation were examined using immunohistochemistry with an anti-IgG and anti-ED-1 antibody. The immunohistochemical analysis revealed that the area of edema formation and inflammation was remarkably reduced in animals with PPI. The muscle function was flaccid in both groups immediately after injury. However, at 3 and 8 days after injury, a significant improvement was observed in the PPI group. These results suggest that PPI is a strong candidate for drug treatment of spinal cord injury. PMID- 14753481 TI - Multiple hyperbaric oxygenation (HBO) expands the therapeutic window in acute spinal cord injury in rats. AB - Hyperbaric oxygenation (HBO) therapy has been reported to improve neurological recovery following spinal cord injury (SCI). In the present study, we examined whether multiple HBO expands the therapeutic window for acute SCI. Single HBO (2.8 ATA, 1 hour) treatment was used at 30 minutes, 3 hours, and 6 hours following SCI, and serial HBO treatment (once daily for 1 week) at 6 hours and 24 hours post-injury. Mild SCI was induced by adjusting the height for a weight drop insult (10 g) to 6.25 mm above the exposed spinal cord. The group of animals receiving a single HBO intervention beginning at 30 minutes and 3 hours, or serial HBO treatment starting at 6 hours following the injury had a significantly better neurological recovery than animals with SCI only. The results of this study demonstrate that multiple HBO expands the therapeutic window for acute SCI to 6 hours after injury, further that serial HBO administration is superior to single HBO therapy. PMID- 14753480 TI - An L-type calcium channel blocker, nimodipine influences trauma induced spinal cord conduction and axonal injury in the rat. AB - The influence of the potent L-type Ca[2+] channel antagonist Nimodipine on spinal cord evoked potentials (SCEP) and axonal injury following trauma to the spinal cord was examined in a rat model. Spinal cord injury (SCI) was produced by an incision into the right dorsal horn of the T10-11 segments under urethane anaesthesia (1.5 g/kg, i.p.). SCEPs were recorded by epidural electrodes placed over the T9 (rostral) and T12 (caudal) segments after stimulation of the right tibial and sural nerves. SCI induced a pronounced decrease of the SCEP negative amplitude in the rostral (T9) recordings immediately after trauma. Axonal injury seen as degradation of myelin basic protein (MBP) immunostaining and myelin vesiculation at the ultrastructural level was most pronounced at 5 h. Continuous administration of Nimodipine (2 microg/kg/min, i.v.) from 30 min prior to injury until sacrifice markedly attenuated the changes in SCEP amplitude and latency. Axonal damage, loss of MBP, and myelin vesiculation were much less evident in the nimodipine treated traumatised rats. These observations suggest that Ca[2+] channels play an important role in the trauma induced alterations in SCEP and axonal injury, and indicate a therapeutic value of Ca[2+] blockers in SCI. PMID- 14753482 TI - The International Surgical Trial in Intracerebral Haemorrhage (ISTICH). AB - At the XIth International Brain Oedema Symposium we reported that 208 patients had been randomized in this trial from 65 centres. At the time of submission of this manuscript we have randomized 985 patients from 107 centres including Japan. The trial will continue randomising patients until the end of February 2003. The objective is to determine whether "Early Surgical Intervention" or "Initial Conservative Treatment" is the best option in patients with spontaneous supratentorial Intracerebral haemorrhage (ICH). Patients with trauma, tumours, aneurysms and angiographically proven arteriovenous malformations are excluded. Ascertainment logs from the various centres have revealed substantial variation in the rate of operation for ICH from 2% in Hungary to 90% in Lithuania. The results will remain blinded until all 6 month follow-up information has been obtained. It is anticipated that these results will therefore be known towards the end of 2003. The pooled results have shown that patients with larger Haematomas and depressed consciousness are more likely to end up with a worse outcome. PMID- 14753483 TI - Chronological changes of perihematomal edema of human intracerebral hematoma. AB - Recent investigations have indicated the importance of secondary brain damage in the pathophysiology of intracerebral hemorrhage (ICH), which includes ischemic brain damage and edema formation around a hematoma. The purpose of the current study is to investigate chronological changes of perihematomal edema in patients with human ICH and also the correlation between volume of perihematomal edema and neurological status. Fourteen patients with medium-sized putaminal hemorrhage (29.4 +/- 13.2 ml) without hematoma enlargement were included in this study. To investigate chronological changes of perihematomal edema, we performed CT scans prospectively on the day of hemorrhage and repeated them on days 1, 3, 7, 14, and 28. We evaluated the patients neurologically using the NIH stroke scale on the day a CT scan was performed. The volume of perihematomal edema in human ICH increased rapidly three days after hemorrhage and the score on the NIH stroke scale showed a deterioration. The volume of perihematomal edema then increased slowly until day 14 after hemorrhage, and decreased thereafter. Despite progression of perihematomal edema, the score on the NIH stroke scale improved gradually after day 3. PMID- 14753484 TI - Thrombin preconditioning upregulates transferrin and transferrin receptor and reduces brain edema induced by lysed red blood cells. AB - Pretreatment with a low dose of thrombin reduces brain edema after both hemorrhagic and ischemic stroke. We call this phenomenon thrombin preconditioning (TPC) or thrombin-induced brain tolerance. The present study examines whether TPC can attenuate the brain edema induced by lysed red blood cells (RBCs) to determine whether thrombin production early in an intracerebral hemorrhage (ICH) might alter potentially injurious events associated with clot resolution. It also examines whether TPC might be protective by altering iron handling within the brain, particularly through modulating transferrin (Tf) and transferrin receptor (TfR) levels. Brain edema was measured by wet/dry weight. Western blot analysis and immunohistochemistry were used for Tf and TfR measurements. We found that TPC reduces lysed RBC-induced brain edema and upregulates both Tf and TfR levels in the brain. Thrombin formation after an ICH may be part of a signaling cascade that acts to limit potentially injurious events associated with clot resolution through altering iron-handling proteins. PMID- 14753485 TI - Effect of hypothermia on brain edema formation following intracerebral hemorrhage in rats. AB - Brain edema is an important clinical complication of intracerebral hematoma (ICH). Hypothermia has been employed as an effective neuroprotective treatment on cerebral ischemia and contusional brain injuries. In this study, we attempted to evaluate the effects of mild hypothermia (35 degrees C) on brain edema formation at 48 hours after ICH in the rat. Brain hypothermia started at 6, 12 and 24 hours after the induction of hematoma (HT6; n = 6, HT12; n = 11, HT24; n = 6). Brain water contents in the basal ganglia was significantly reduced in the rats treated with mild hypothermia compared with the normothermic rats (NT; 82.0 +/- 0.04% vs. HT6; 78.6 +/- 0.09%: p < 0.01, HT12; 79.7 +/- 0.05%.: p < 0.01, HT24; 79.7 +/- 0.05%: p < 0.01). Differences in the brain water content were not significant among the hypothermic subgroups. The BBB disruption to Evan's blue was significantly reduced with hypothermia compared with the normothermic rats in the ipsilateral basal ganglia (42.3 +/- 4.0 vs. 23.0 +/- 5.2 ng/g wet tissue; p < 0.05). Hypothermic treatment tended to inhibit the accumulation of polymorphonuclear (PMN) leukocytes in the lesion compared with the normothermic treatment (0.78 +/- 0.13 vs. 0.51 +/- 0.16 deltaAbs/mg tissue). This study indicates that hypothermic treatment significantly reduces the brain edema formation after ICH in rats. PMID- 14753486 TI - Effect of delayed argatroban treatment on intracerebral hemorrhage-induced edema in the rat. AB - Studies indicate that thrombin plays an important role in intracerebral hemorrhage (ICH) induced edema formation. However, the time window for administration of a thrombin inhibitor to reduce ICH-induced edema is unknown. Nor is it known whether this time window extends beyond the period when a thrombin inhibitor might exacerbate rebleeding. This study examines whether a thrombin inhibitor, argatroban, can reduce edema formation following intracerebral infusion of 100 microl of blood in the rat, the therapeutic time window for argatroban, and whether argatroban promotes rebleeding. Intracerebral injection of argatroban 3 hours after ICH caused a significant reduction in edema measured at 48 hours. The systemic administration of argatroban (0.9 mg/h) starting 6 hours after ICH also significantly reduced edema formation. There was no protection when the onset of argatroban administration was delayed to 24 hours after ICH. Argatroban did not increase collagenase-induced hematoma volume when given into the clot after 3 hours or given systemically at 6 hours. Our data suggest argatroban may be an effective therapy for ICH-induced edema. PMID- 14753487 TI - Overexpression of interleukin-1 receptor antagonist reduces brain edema induced by intracerebral hemorrhage and thrombin. AB - Recent studies indicate that inflammatory reaction occurs around hematoma after intracerebral hemorrhage (ICH). In this study the authors examine the hypothesis that overexpression of IL-1ra in the brain attenuate brain edema formation after ICH. Adenoviruses expressing IL-1ra (Ad.RSVIL-1ra) or LacZ (Ad.RSVLacZ) or saline were injected into the lateral ventricle. On the fifth day after virus injection, 100 microl of autologous blood or 5 U thrombin was infused into the right basal ganglia. Rats with ICH were killed 24 or 72 hours later for measurement of brain water content. Thrombin-treated rats were killed 24 hours later for edema measurements and an assessment of polymorphonuclear leukocyte (PMNL) infiltration by myeloperoxidase (MPO) assay. Compared with control groups, Ad.RSVIL-1ra treated rats had less brain edema formation in the ipsilateral basal ganglia 3 days after ICH (81.5 +/- 0.3% compared with 83.4 +/- 0.4% and 83.3 +/- 0.5% in control animals). Ad.RSVIL-1ra treated rats had also less brain edema following thrombin injection. The reduction of brain edema induced by thrombin was involved in the reduction of PMNL infiltration in basal ganglia, as assessed by MPO assay. Adenovirus-mediated overexpression of IL-1ra attenuated brain edema formation following ICH, perhaps by reduction of thrombin-induced brain inflammation. PMID- 14753488 TI - The role of haematoma aspiration in the management of patients with thalamic haemorrhage: a pilot study with continuous compliance monitoring. AB - OBJECTIVE: Thalamic brain haemorrhage is a common disabling and potentially fatal condition. Management is mainly supportive and very rarely do neurosurgeons have to resort to evacuation of the haematoma. We hypothesised that haematoma aspiration may help to improve the outcome of the group of patients with thalamic haemorrhage. MATERIAL AND METHODS: Patients with thalamic haemorrhage were selected on clinical group (intraventricular haemorrhage, hydrocephalus and haematoma size > 15 ml) for intracranial pressure (ICP) monitoring and management in the intensive care unit (ICU). A Spiegelberg double lumen intraventricular balloon catheter was inserted as a fluid-filled catheter on the side of the haematoma. Data of ICP, arterial blood pressure (ABP), cerebral perfusion pressure (CPP) and compliance were collected at one-minute intervals. Hourly averages were used for analysis. RESULTS: 94,448 set of data were collected from eight patients with thalamic haemorrhage. Significant reduction in ICP and improvement in intracranial compliance were detected. In patients with haematoma aspiration, intracranial compliance was improved within 48 hours of the procedure whereas ICP did not. CONCLUSIONS: This pilot study showed that haematoma aspiration in patients with thalamic haemorrhage could improve intracranial compliance. Whether this could be translated into clinical outcome benefits requires further study in a larger number of patients. PMID- 14753489 TI - Acute decrease of cerebrocortical microflow and lack of carbon dioxide reactivity following subarachnoid haemorrhage in the rat. AB - Experiments were designed to study the extent, duration and severity of the impairment of cerebrocortical microciroflow during acute ischemia following experimental SAH in the rat. Twenty five male, adult anesthetized and mechanically ventilated Wistar rats were used. SAH was induced by perforation of the middle cerebral artery (MCA) using intravascular filament. Cerebrocortical microflow (LDF) was recorded bilaterally in the territory supplied by the MCA at normocapnia before SAH and up to 180 min thereafter. Reactivity of microcirculation to CO2 was also studied. In order to further explain mechanisms of post-SAH microcirculatory changes, L-arginine--a precursor of NO and 17 octadecynoic acid (17-ODYA)--an inhibitor of the enzymes of cytochrome P-450 family were administered. SAH resulted in acute decrease of microflow on both sides although during the first 20 min this effect was much better pronounced on the side ipsilateral to ruptured MCA (p < 0.05). Pretreatment with L-arginine or 17-ODYA didn't improve microflow after SAH. On the contrary, in rats pretreated with 17-ODYA LDF on the side ipsilateral to bleeding significantly deteriorated (p < 0.05 vs. control group at all times beginning 10 min after SAH). Following SAH the impairment of CO2 reactivity was also observed. PMID- 14753490 TI - Heat shock proteins expression in brain stem after subarachnoid hemorrhage in rats. AB - The pathogenesis of brain damage after subarachnoid hemorrhage (SAH) especially at molecular or gene level remains unclear. We used complimentary deoxyribonucleic acid (cDNA) macroarray technique and compared gene expression in brain stem after experimental SAH in rats. The upregulation of several heat shock proteins (HSPs) demonstrated by cDNA array was further confirmed by Western blotting. The expressions of 9 genes were upregulated 30 minutes or 2 days after SAH. They included four upregulated HSPs: HSP90alpha, HSP60, HSP27, and HSP10. Western blotting demonstrated increases in the HSP27 and HSP10 proteins on Day 2. SAH enhanced the induction of several HSP mRNAs in the brainstems, even though the functions of these HSPs after SAH remain unclear. PMID- 14753491 TI - Apoptosis, blood-brain barrier, and subarachnoid hemorrhage. AB - This study was undertaken to investigate the role of apoptosis in the integrity of blood-brain barrier (BBB) in subarachnoid hemorrhage (SAH). BBB permeability changes were examined and found increased on day 7 in a double hemorrhage rat model using Evans blue dye. The BBB permeability increase is coincidental to brain microvascular endothelial cell apoptosis (expression of caspase-8 and -9) occurring on Day 7. However, caspase-8 and caspase-9 inhibitors failed to protect the BBB. Considering that treatment did not completely inhibit apoptosis in brain microvascular endothelial cells, higher doses, earlier and/or multiple applications, and, possibly, more potent caspase inhibitors may be needed. PMID- 14753492 TI - Focal brain edema and natriuretic peptides in patients with subarachnoid hemorrhage. AB - Central salt wasting syndrome may be caused by pathological increases in serum natriuretic peptides after subarachnoid hemorrhage (SAH). However, it is unclear as to why the serum concentration of atrial natriuretic peptide (ANP) or brain natriuretic peptide (BNP) increases in the subacute phase of SAH. The present study was designed to assess the correlation between focal brain edema and serum concentration of ANP or BNP in patients with SAH. Focal brain edema was found in 8 SAH-patients and peaked between days 4 and 7 of SAH. The mean serum ANP and BNP levels in patients with focal brain edema were significantly higher than those in patients without focal brain edema between days 4 and 14 of SAH. These results suggest that focal brain edema might correlate with increased levels of ANP and BNP in the subacute phase of SAH. PMID- 14753493 TI - Aquaporin 1 and aquaporin 4 expression in human brain after subarachnoid hemorrhage and in peritumoral tissue. AB - Aquaporins (AQPs) are a protein family of water channels which facilitate the water flux through the plasmatic membranes. The expression of AQPs has been described in rat brain by several studies. Despite recent reports that have shown an over-expression of AQP1 and 4 in human tumoral cells, little is known about AQP expression in human brain. The purpose of this study was to investigate the expression of AQP1 and AQP4 in human brain after subarachnoid hemorrhage (SAH) and in peritumoral tissue by western blot and immunohistochemistry. The results showed a marked increase of the expression of AQP1 and AQP4. This over-expression occurred on the astrocytic processes and polarization on astrocytic end-feet was lost. No expression was observed on neuronal cells. This study is the first demonstration of the induction of AQP1 and AQP4 on reactive astrocytes in an acute brain injury, such as SAH. These results reinforce the hypothesis that AQPs may be involved in the dynamics of brain edema formation or resolution. Further studies are needed to understand their functional role. PMID- 14753494 TI - Aquaporin-1 expression in human glial tumors suggests a potential novel therapeutic target for tumor-associated edema. AB - Aquaporins are membrane proteins involved in water transport in many fluid transporting tissues. The objective of this study was to investigate the expression of aquaporins in malignant glial tumors associated with cerebral edema. Eighteen human glial tumors were obtained from the UCSF Neurosurgery Tissue Bank. Aquaporin-1 (AQP1) expression was evaluated via Western blot and immunohistochemistry. Intense upregulation of AQP1 expression was found in all glioblastomas. The robust expression of aquaporins in glioblastomas suggests a pathologic role for these membrane water channels, and raises the possibility that selective AQP inhibition might offer a new therapeutic option for tumor associated cerebral edema. PMID- 14753495 TI - A thrombin inhibitor reduces brain edema, glioma mass and neurological deficits in a rat glioma model. AB - Although thrombin is a critical enzyme in the coagulation cascade, it has become apparent that it has many other effects. Thus, it may induce brain edema formation, angiogenesis and cell proliferation. Because of the importance of these three factors in the extremely poor prognosis of glioma patients, the present study examined the role of thrombin in that disease state. We found that thrombin activity is increased in a rat glioma model and thrombin positive cells were present in the tumor. Anti-thrombin treatment with argatroban reduced brain edema, tumor growth, and tumor-related neurological deficits. Our results suggest that thrombin is a new target for glioma treatment. PMID- 14753496 TI - Tissue reconstruction process in the area of peri-tumoural oedema caused by glioblastoma--immunohistochemical and graphical analysis using brain obtained at autopsy. AB - BACKGROUND: In the area of peri-tumoural oedema, proteolytic agents derived from the tumour cause tissue degradation, which promotes tumour cell invasion. METHOD: We investigated the biological processes in the area of peri-tumoural oedema, using a brain obtained at autopsy from a patient who died from glioblastoma. Immunohistochemistry was performed to detect vascular endothelial growth factor (VEGF), c-myc, p53, paternally expressed gene-3 (PEG-3), transforming growth factor beta (TGFB), and tumour necrosis factor alpha (TNFA). The data were translated into colour graphics and the localization of these proteins was analyzed. FINDINGS: In the area of peri-tumoural oedema, Ki-67 and p53 positive cells were observed with TGFB expression. Moreover, c-myc, PEG-3, VEGF, and TNFA were also expressed strongly in the glial cells or extra-cellular spaces in the area of peri-tumoural oedema. INTERPRETATION: These data suggest that in the area of peri-tumoural oedema, tissue reconstruction processes take place with concomitant anti-tumour activities. The expression of c-myc, VEGF, and TNFA in the area of peri-tumoural oedema may indicate that these proteins are not utilized for tumour growth, but may be used to guard the brain against tumour invasion. Peri-tumoural oedema does not only indicate the tissue damage caused by tumour, but many tissue reconstruction processes take place in these areas against tumour cell invasion. PMID- 14753497 TI - Radiation necrosis and brain edema association with CyberKnife treatment. AB - The CyberKnife (CK) is a frameless and image guided robotic controlled instrument for stereotactic irradiation. The authors studied CK treatment of glioma and glioblastoma, and analyzed frequency and risk factors of radiation necrosis. Of 61 patients with glioma and glioblastoma treated with CyberKnife, four patients showed symptomatic radiation necrosis. All of these patients were treated with stereotactic radiotherapy, varying from 3 to 6 fractions without previous radiation therapy. Two patients required necrotomy through craniotomy. Two patients were treated conservatively. Our four patients with radiation necrosis were not specific in terms of tumor volume and dose delivery. Glioma cells invade the normal brain tissue and over-radiation to this intermingling area is one of the risk factors for injury to normal endothelial cells. The homogeneity of the maximum dose area is an important factor to reduce over radiation to the normal brain parenchyma. The dose volume effect has been discussed in terms of risk factor; however, the number of fractions and dose per fraction should be considered to avoid radiation necrosis. We consider that conformal treatment with inverse algorism, fractionated stereotactic radiotherapy and precise anatomic targeting reduce the risk of radiation necrosis. PMID- 14753498 TI - Vascular endothelial growth factor expression in pituitary adenomas. AB - Vascular endothelial growth factor (VEGF) is known to be a mediator of angiogenesis and vascular permeability. A cystic component and hemorrhage are often found in pituitary adenomas. In the present study we assess the VEGF expression based on immunohistochemical examinations in 48 pituitary adenomas. All the adenomas showed some VEGF immunoreactivity mainly in the cytoplasm of tumor cells. Of the 48 adenoma-cases, 16 cases had a strong VEGF immunoreactivity, 26 cases had a moderate one, and 6 cases had a weak one. On the MR images, a cystic component was found in 16 cases (33.3%), and a hemorrhage was found in 18 cases (37.5%). The VEGF immunoreactivity had a significant relationship with the cystic component but neither the hemorrhage, size, recurrence, or HE classification. These findings suggest that VEGF might play a potential role in the pathogenesis of cystic formation in pituitary adenomas. PMID- 14753499 TI - Aquaporin-1 deletion reduces osmotic water permeability and cerebrospinal fluid production. AB - Aquaporin-1 (AQP1) is a water channel that is strongly expressed at the ventricular-facing surface of choroid plexus epithelium. Using wildtype and AQP1 null mice, we developed novel methods to compare the water permeability in isolated choroid plexus, and cerebrospinal fluid (CSF) production in living mice. Osmotically-induced water transport was rapid in freshly isolated choroid plexus from wildtype mice as measured by a spatial-filtering optical method, and reduced by 5-fold by AQP1 deletion. CSF production, an isosmolar fluid secretion process, was measured by a dye dilution method involving fluid collections using a second microneedle introduced into the cisterna magna. CSF production in wildtype mice was (in microl/min) 0.37 +/- 0.04 microl/min (control), 0.16 +/- 0.03 microl/min (acetazolamide-treated) and 1.14 +/- 0.15 microl/min (forskolin-treated), and reduced by up to 25% in AQP1 null mice. The impaired CSF production in AQP1 null mice provides direct functional evidence for the involvement of AQP1 in CSF formation. PMID- 14753500 TI - Hysteresis of the cerebrospinal pressure-volume curve in hydrocephalus. AB - The objective was to study the displacement of the cerebrospinal fluid pressure volume curve during the descent relative to the ascent of intracranial pressure recorded during the cerebrospinal fluid constant rate infusion test. This phenomenon can be interpreted as the hysteresis of the pressure-volume curve. The cerebrospinal fluid dynamics were tested in fifty-eight patients with clinical symptoms of hydrocephalus. After finished infusion, ICP was recorded until it returned to steady state level. Pressure-volume curves were plotted separately for ascending and descending phases of the test. The parameters of CSF compensation were estimated on the basis of mathematical mono-exponential model of CSF circulation. The pressure-volume curve post-infusion was visibly shifted upward in 69% of tests. Those who demonstrated the upward shift of the pressure volume curve had greater an elastance coefficient of the cerebrospinal space (with shift: E1 = 0.26 +/- 0.14; without shift: E1 = 0.17 +/- 0.06; p < 0.05). Magnitude of the shift was positively correlated with pulse amplitude of ICP (r = -0.763; p < 0.0001). The accuracy of clinical examination of the pressure-volume compensatory reserve, which take into account both compression and decompression phase of the study, may be affected by this phenomenon. PMID- 14753501 TI - Is decreased ventricular volume a correlate of positive clinical outcome following shunt placement in cases of normal pressure hydrocephalus? AB - It is well known that in patients with communicating hydrocephalus or normal pressure hydrocephalus, ventricular volume decreases following implantation of differential pressure valved shunts. Hydrostatic valves (Miethke dual-switch valves) were implanted in 60 normal pressure hydrocephalus patients at Unfallkrankenhaus Berlin between September 1997 and September 2001. One year postoperatively, these patients underwent CT scan, and their ventricular size was ascertained using the Evans index. Although 77% of these patients showed no postoperative change in ventricular volume, 65% nonetheless showed good to excellent clinical improvement, 13% satisfactory improvement and 22% no improvement. A moderate reduction in ventricular size was observed in 17% of the patients in our cohort. 40% of these patients showed good to excellent clinical improvement, 20% satisfactory improvement, and 40% unsatisfactory improvement. A marked reduction in ventricular size was observed in 6% of our patients. Of these latter patients, 50% showed good to excellent outcomes, while 50% had unsatisfactory outcomes. The favorable outcomes following implantation of a hydrostatic shunt in patients with normal pressure hydrocephalus did not correlate with decreased ventricular volume one year after operation. Better clinical outcomes were observed in patients with little or no alteration in ventricular size than in patients with a marked decrease in ventricular size. Postoperative change in ventricular volume should be assessed differently for patients with normal pressure hydrocephalus than in patients with hypertensive hydrocephalus. PMID- 14753502 TI - The ICP-dependency of resistance to cerebrospinal fluid outflow: a new mathematical method for CSF-parameter calculation in a model with H-Tx rats. AB - The international accepted calculation methods concerning the cerebrospinal fluid dynamics proceed from a pressure independent resistance to cerebrospinal fluid outflow. In a new model we focus our attention on the pressure dependency of resistance. In our calculation model we are monitoring the complete pressure course p(t) over the time t during and after the infusion. The comparison of the pressure rise On(p) during the infusion and the descent Off(p) after the infusion in the same pressure level allows to construct all formulas for the compliance C(p) and resistance R(p). The simultaneous measurement of the resistance and complications during a single investigation allows minimizing the patient's exertion. In contrast to the classical methods it is not necessary that the ICP reach a plateau. Our mathematical method diverges with the description of a pressure dependent slope of the function for the resistance from the static examination models. We proved our mathematical method by the use of a dynamic infusion test in ten H-Tx rats without hydrocephalus and five hydrocephalic H-Tx rats. For that we are able to take the non-linearity of the CSF resorption into consideration. PMID- 14753503 TI - Metabolic derangements in interstitial brain edema with preserved blood flow: selective vulnerability of the hippocampal CA3 region in rat hydrocephalus. AB - Hydrocephalus induces interstitial brain edema, which causes neurological deficits, even if the intracranial pressure is maintained within the normal range, and the cerebral blood flow (CBF) does not decline to an ischemic level. The precise mechanisms underlying such edema-induced neuronal dysfunction remain unclear. In the present study, in an attempt to elucidate the metabolic derangements in brain tissue with interstitial edema, we evaluated the changes in CBF and oxidative/glucose metabolism using a rat model of kaolin-induced hydrocephalus. Hydrocephalus was produced in male Wistar rats by intrathecal injection of 0.1 ml aluminum silicate suspension (200 mg/ml) via the cisterna magna. CBF was determined by 14[C]-iodoantipyrine autoradiography. Oxidative metabolism was evaluated by cytochrome oxidase (CYO) histochemistry, and glucose metabolism by hexokinase (HK) histochemistry. CBF declined with the development of hydrocephalus, but did not reach an ischemic level. The CYO activity was diffusely depressed in both the cortex and hippocampus. The HK activity was preserved at the early stage of hydrocephalus. At the advanced stage, the HK activity was reduced in the hippocampal CA3 region first, and diffusely thereafter. In conclusion, interstitial brain edema impairs oxidative metabolism even at the early stage of hydrocephalus, and shifts the metabolism to anaerobic glycolysis despite a preserved CBF. Impairment of glucose metabolism was first observed in the CA3 region, suggesting that the CA3 is metabolically vulnerable, and CA3 dysfunction may contribute to the memory deficits seen in hydrocephalus. PMID- 14753504 TI - MRI analysis of hydrocephalus associated with acoustic neurinoma. AB - We investigated the hydrocephalus in 24 patients associated with acoustic neurinoma. We found the high incidence of homo-lateral ventricular dilatation to the side of the acoustic neurinoma. Utilizing magnetic resonance imaging, the diameter of the tumor parallel to the pyramidal bone, diameter of the tumor perpendicular to the pyramidal bone, grade of the 4th ventricle deviation, and the shape of the tumor (round or oval) were analyzed. Ten (42%) of the 24 patients with acoustic neurinoma were found to have hydrocephalus. Seven (70%) of the 10 patients with hydrocephalus exhibited asymmetrical lateral ventricle dilatation: in all cases the lateral ventricle in the hemisphere homolateral to the acoustic neurinoma was larger than that of the contralateral side. The hydrocephalus was not related to the grade of the 4th ventricle deviation but rather to the diameter of the tumor parallel to the pyramidal bone (p < 0.01). The diameter of the tumor parallel to the pyramidal bone was also related to the asymmetrical lateral ventricular dilatation (p < 0.05). PMID- 14753505 TI - Non-invasive opening of BBB by focused ultrasound. AB - Blood brain barrier (BBB) is a major barrier for delivering therapeutic agents in the brain. In this study we investigated the feasibility of open the BBB by using focused ultrasound. Rabbit brains were exposed to pulsed focused ultrasound while injecting ultrasound contrast agent containg microbubbles intravenously. The BBB opening was measured after the sonications by injecting MRI contrast agent i.v. and evaluating the local enhancement in the brain. Low ultrasound powers and pressure amplitudes were found to cause focal enhancement. Before sacrificing the animals trypan blue was also injected i.v.. After the sacrifice of the animals blue spots were found in the brain in the sonicated locations. This method may have potential for targeted delivery of macromolecules in the brain. PMID- 14753506 TI - Quantitative analysis of hyperosmotic and hypothermic blood-brain barrier opening. AB - Hyperosmotic opening of the blood-brain barrier (BBB) by mannitol is being used to enhance drug transport in human brains. Recently, cooling of the solution has been reported to have potential to open the BBB. However, the mechanism in barrier opening and closure remains elusive. We studied the rapid changes in cerebrovascular permeability after hyperosmotic and hypothermic BBB opening in rats, and then demonstrated that the Na+/Ca++ exchange blocker (KB-R7943) prolongs opening. BBB opening was attained by using intra-arterial infusion of hyperosmotic mannitol (1.6 M) and 1.1 M mannitol (which is less hyperosmotic than commonly used mannitol) at 4 degrees in Sprague-Dawley (SD) rats. To measure the changes in cerebrovascular permeability, perfusate-containing [14C]-sucrose was infused intra-arterially at different time points following hyperosmotic and hypothermic stress. Cerebrovascular permeability was then measured with the in situ brain perfusion technique. 1.6 M Mannitol produced opening of the BBB but the duration of the opening was less than 30 minutes. Use of 1.1 M Mannitol at 4 degrees indicated the same results. We then investigated the effect of a Na/Ca ion exchange blocker (KB-R7943) in both hyperosmotic and hypothermic BBB opening. KB-R7943 extended BBB opening up to 30 min without affecting the peak level of BBB permeability at 5 minutes. Our findings represent important experimental information regarding pharmacological manipulation of BBB opening. The possibility of prolonging the transient opening of the BBB has major clinical implications. PMID- 14753507 TI - Chronic adrenomedullin treatment improves blood-brain barrier function but has no effects on expression of tight junction proteins. AB - We previously found that the production of adrenomedullin (AM) is one magnitude higher in cerebral endothelial cells (CECs) than in the peripheral endothelium and the AM concentration in the cerebral circulation is significantly higher than in other tested parts of the circulation. We also showed that CECs express AM receptors, and AM as an autocrine hormone is important to regulate the intracellular cAMP level in CECs. Further we reported that acute AM treatment has cAMP-like effects on specific BBB functions: AM decreased endothelial fluid phase endocytosis, activated the P-glycoprotein, increased transendothelial electrical resistance (TEER) and reduced endothelial permeability for sodium fluorescein, which suggests a tightening of intercellular junctions. In the present study, we found chronic AM exposure also increased TEER. In contrast, we could not detect significant effect of AM on the expression of tight junction proteins (claudin-1, occludin and zonula occludens-1). While not affecting expression of tight junction proteins, chronic AM treatment may influence the localization of these proteins which has been reported to correlate with functional changes of the BBB without a change in protein expression. PMID- 14753508 TI - Characteristic phosphorylation of the extracellular signal-regulated kinase pathway after kainate-induced seizures in the rat hippocampus. AB - Extracellular signal-regulated kinase (ERK) pathways play a crucial role in cell growth and long-lasting neuronal plasticity. Several studies have shown that phosphorylated-ERK (p-ERK) significantly increases after kainic acid (KA) administration. However, little or no information is available about the spatial distribution of p-ERK after KA-induced seizures. We herein show that KA-induced seizures significantly increase p-ERK in both neurons and astrocytes in rat brain using Western blots and immunohistochemistry. A strong immunoreactivity for p-ERK was induced in the dentate hilar neurons and CA3 neurons 30 mins and 6 hrs after KA injection. In addition, immunoreactivity for p-ERK was seen in astrocytes 6 hrs after KA injection. 72 hrs after KA injection, all pyramidal neurons had died. These findings suggest that the ERK pathway participates in the KA-induced neurotoxicity in the rat hippocampus. PMID- 14753509 TI - Highly polysialylated neural cell adhesion molecule (PSA-NCAM) positive cells are increased and change localization in rat hippocampus by exposure to repeated kindled seizures. AB - The highly polysialylated neural cell adhesion molecule (PSA-NCAM) is involved in migration of neural stem cells as well as in neural plasticity. Immunoreactive PSA-NCAM expression was examined in rats with repeated exposure to amygdaloid kindled generalized seizures (GS). The number of PSA-NCAM positive cells in the bilateral dentate gyrus (DG) increased significantly from GS. Although the total number of positive cells was not significantly different between animals with 3 times GS (3 GS) and 30 times GS (30 GS), in the latter group a greater number of positive cells was observed in the outer granule cell layer (GCL) and a marked extension of immunopositive dendrites to the molecular layer. These observations indicate that increased migration of newly generated cells as well as plastic changes of preexisting neural cells occur in response to recurrent GS. This may contribute to an abnormal reconstruction of the synaptic network in the hippocampus and, thus, epileptogenicity from kindling. PMID- 14753510 TI - Continuous assessment of cerebral autoregulation: clinical and laboratory experience. AB - The method for the continuous assessment of cerebral autoregulation using slow waves of MCA blood flow velocity (FV) and cerebral perfusion pressure (CPP) or arterial pressure (ABP) has been introduced seven years ago. We intend to review its clinical applications in various scenarios. Moving correlation coefficient (3 6 min window), named Mx, is calculated between low-pass filtered (0.05 Hz) signals of FV and CPP or ABP (when ICP is not measured directly). Data from ventilated 243 head injuries and 15 patients after poor grade subarachnoid haemorrhage, 38 patients with Carotid Artery stenosis, 35 patients with hydrocephalus and fourteen healthy volunteers is presented. Good agreement between the leg-cuff test and Mx has been confirmed in healthy volunteers (r = 0.81). Mx also correlated significantly with the static rate of autoregulation and transient hyperaemic response test. Autoregulation was disturbed (p < 0.021) by vasospasm after SAH and worse in patients with hydrocephalus in whom CSF circulation was normal (p < 0.02). In head injury, Mx indicated disturbed autoregulation with low CPP (< 55 mmHg) and too high CPP (> 95 mmHg). Mx strongly discriminated between patients with favourable and unfavourable outcome (p < 0.00002). This method can be used in many clinical scenarios for continuous monitoring of cerebral autoregulation, predicting outcome and optimising treatment strategies. PMID- 14753511 TI - Spontaneous intracranial hypotension (SIH): the early appearance of urinary bladder activity in RI cisternography is a pathognomonic sign of SIH? PMID- 14753512 TI - Changing needs for appropriate excreta disposal and small wastewater treatment methodologies or The future technology of small wastewater treatment systems. AB - Recent developments will strongly influence the design and utilization of small wastewater treatment systems in the future, e.g. population increases in non sewered rural areas and developing countries; increasing impairment of surface water quality; the construction of occupied high-rise buildings in metropolitan areas; the development of planned but somewhat isolated communities, growing shortages of water that mandate reuse of wastewaters. It is well known that there is a very strong linkage between wastewater disposal methods in rural areas and developing countries and the general health of the population. These problems could be greatly reduced or prevented by the utilization of well known excreta disposal and small wastewater treatment system technologies, but the development of more innovative on-site systems is needed. It is expected that future environmental and public health pressures in developed countries will require increasingly stringent effluent limitations for small and on-site wastewater disposal systems, based primarily on nutrient discharges. Both on-site and small scale technologies are available for the more stringent requirements, but innovative and more economical designs are needed for wide-spread acceptance. Water reuse should be a consideration for the designs of these systems. Implementation and utilization of well known technologies are needed, but the obstacles are often more social and political than economical. PMID- 14753513 TI - Small water and wastewater systems: pathways to sustainable development? AB - Globally we are faced with billions of people without access to safe water and adequate sanitation. These are generally located in developing communities. Even in developed communities the current large scale systems for supplying water, collecting wastewater and treating it are not environmentally sustainable, because it is difficult to close the cycle of water and nutrients. This paper discusses the advantages of small scale water and wastewater systems in overcoming the difficulties in providing water and wastewater systems in developing communities and in achieving sustainability in both developed and developing communities. Particular attention is given to technology and technology choice, even though technology alone does not provide the complete answer. Disadvantages of small scale systems and how they may be overcome are discussed. PMID- 14753514 TI - Scientific basis for the design of small activated sludge systems. AB - The study presents an evaluation of the oxygen requirement and sludge production, modeled in terms of fundamental processes reflecting different biochemical transformations in activated sludge systems. Modern modeling concepts define substrate utilization and endogenous decay as major processes requiring final electron acceptors. Substrate and sludge components may be defined in terms of different parameters. Chemical oxygen demand (COD) and biochemical oxygen demand (BOD5) are the traditional substrate parameters. COD also serves to define biomass along with volatile suspended solids (VSS) parameter. Estimation of oxygen requirement and sludge production for aerobic activated sludge systems covering basic biochemical processes are defined for different substrate and sludge parameters, considering that each calculation is associated with a different stoichiometry on the basis of a selected parameter set. The German regulations are examined and the biochemical bases of the coefficients in the regulatory expressions are set in terms of fundamental model constants. PMID- 14753515 TI - Innovative technologies for decentralised water-, wastewater and biowaste management in urban and peri-urban areas. AB - Avoiding the comingling of water flows coming from different sources and thus obtaining flows with a very low dilution factor is the first and major step key to technical solutions for adequate treatment of household wastewaters. Through their decentral structure and effective recovery of water, energy and fertiliser these systems can be highly cost efficient. Fresh water consumption can be reduced by up to 80% while nutrients can be recovered to a large extent. Source control is also advantageous for hygienic reasons: low volumes are far easier to sanitise. Source separation technology in municipal waste water treatment does often lead decentralised or semicentral systems. The first essential step is the separate collection and treatment of toilet waste in households, which contains almost all pathogens and nutrients. New toilet systems with very low dilution factors, ranging from vacuum- through urine sorting to dry toilets, have been introduced in several projects and proven feasible. New ideas such as the black- and greywater cycle systems are presently under research at the Technical University Hamburg Harburg. Such modular, integrated and small scale systems are only possible through recent advances in membrane technology and, due to their small scale, do have the potential to be installed in densely populated regions. These technologies are options for following the principles of ecological sanitation, to contain, to sanitise and to reuse also in urban areas (EcoSanRes, 2003). PMID- 14753516 TI - Sludge management of small water and wastewater treatment plants. AB - "Disposal or reuse?" is the big question in respect to sewage and water sludges. This question is even more decisive for small plants, because the achievement of the advanced treatment necessary for disposal is much more difficult in small units than in big ones. The paper discusses this question with main focus on small WWTPs and gives a short overview of the specifics of sludge treatment in small units as well as of outlets for sludge from water treatment plants. In general, small WWTPs do not have a better sludge quality than larger ones, but the economic pressure to realise reuse is higher on them than on large plants. PMID- 14753517 TI - Developments in the application of chemical technologies to wastewater treatment. AB - Increasing demand for high degree of treatment as well as existence of highly resistant organics in wastewaters such as micropollutants has caused wider and ever increasing use of chemical treatment processes. In this paper developments in the chemical treatment as applied to mostly industrial wastewater in the last two decades are summarized and discussed. Two purely chemical means of treatment, chemical oxidation and chemical precipitation were considered and proven and widely used methods of treatment based on these principles were delineated. The review covers theoretical development, application areas and application bases. A brief evaluation of the state of development, advantages, drawbacks and need for further research was given for the processes reviewed. PMID- 14753518 TI - Centralised versus decentralised sewage systems: a comparison of pathogen and nutrient loads released into Sydney's drinking water catchments. AB - Data collected from centralised and decentralised sewage treatment plants throughout Sydney's drinking water catchments was used to calculate the relative catchment loads of Cryptosporidium, enteric viruses, nitrogen and phosphorus for an initial screening assessment. Loads were assessed at median and 90 percentile values for expected and worst-cases scenarios. The expected scenario in the Sydney drinking water catchments is that decentralised systems (servicing 32,800 people) provide similar total loads to centralised systems (serving 70% of the catchment population) for total phosphorus (37,090 kg x y(-1)), Cryptosporidium (10(11) oocysts x y(-1)) and enteric viruses (9.1 x 10(13) y(-1)), but higher loads of total nitrogen (237,610 vs. 136,740 kg x y(-1)). Decentralised systems, however, were predicted to have higher loads in the worst-case scenario with 620,620 kg x y(-1) TN, 82,040 kg x y(-1) TP, 7.3 x 10(13) Cryptosporidium oocysts x y(-1) and 9 x 10(15) enteric viruses per year. Greater load variability was experienced with decentralised systems, which presumably reflects less reliability in their current operation and maintenance. Overall, catchment water quality is therefore not only affected by sewage disposal methods, but also failure issues. Decentralised system disposal to land may afford a degree of mitigation that can be enhanced, if the degree of failure is reduced. PMID- 14753519 TI - Evaluation of small wastewater treatment systems. AB - Different technologies for the treatment of wastewater from single houses have been evaluated by performing measurements at 14 treatment plants installed at different one-family houses located 35 km south-west of Stockholm. The technologies tested were selected in order to deliver high removal efficiencies with respect to organic matter, nitrogen and phosphorus. They also had to fulfil requirements concerning robustness, nutrient recycling, use of natural resources, economy, user-friendliness and hygiene. The systems can be divided into package plants, urine separating systems, blackwater separating systems and chemical precipitation as a supplement to sand filter beds. All of the systems were able to remove more than 90% of phosphorus and more than 90% of organic matter. Concerning package plants, it is necessary to have reliable dosing equipment and frequent checks to achieve long-term phosphorus removal. The source separation systems require well-informed and motivated users to achieve desirable removal efficiency. Sorting systems seem to be superior with respect to nitrogen removal, even though some of the package plants gave nitrogen removal above 50% during long periods of operation. PMID- 14753520 TI - Joint operation of small wastewater treatment plants in southern Turkey. AB - In this study, joint operation of several small wastewater treatment plants (SWWTPs) by the same operation company is proposed to avoid operational problems such as unqualified personnel, inadequate maintenance and laboratory services. Some case studies from Antalya province of Turkey are presented. Several SWWTPs are operated by the companies formed with the proportional shares of the owners such as ALTAS and TURAS. The performance data of the five treatment plants operated by ALTAS and two treatment plants operated by TURAS are presented. The status of wastewater treatment with emphasis on the small sized plants is also presented and evaluated. The percentage of small settlements served with sewer and wastewater treatment facilities is very low (3%) indicating that high volumes of investment are needed to increase the level of service to those of the developed countries. At present, the total number of the treatment plants in Antalya region is 409 with capacities ranging from 73 to 175 m3 d(-1). Package activated sludge type SWWPs built especially in the coastal regions meet the local effluent standards. PMID- 14753521 TI - A collection and treatment system for organic waste and wastewater in a sensitive rural area. AB - In the municipality of Sund, located in a sensitive rural area in Aland, a demonstration project is now carried out with the overall objective to move the most concentrated fractions of wastewater from the coastal area to a treatment plant situated close to arable land. Blackwater and greywater septic sludge from about twenty households and two tourist camps are treated together with energy rich organic material from a nearby potato-chip factory. The collection concept is based on the use of extremely efficient water-saving toilets, with separate systems for the blackwater and greywater in the households. The collected materials are co-treated in a batchwise aerobic thermophilic treatment process (wet composting process), where the materials reach at least 55 degrees C during a minimum of 10 hours. The dry matter content of the collected material was about 2%. After stabilisation and sanitation (by the temperature rise caused by microbial activity during the treatment process), the compost slurry is utilized as a liquid organic fertilizer on arable land. PMID- 14753522 TI - Application of small community sewer system for improving the quality of the water resource in Korea. AB - An existing SBR plant in a rural area was retrofitted from a conventional fill and-draw system to an intermittent-aeration system for additional nitrogen removal. This study indicated that organic and nitrogen removal efficiency was improved over that before the retrofitting. But effluent phosphorus concentration was increased gradually with the operating time. In the latter period of investigation, phosphorus concentration in effluent was higher than influent. It was regarded that an excessively accumulated phosphorus was released again under the anaerobic conditions of the sludge storage tank. The application of the electro-coagulation process was investigated as an alternative method in order to prevent phosphorus from re-releasing. A laboratory test for electro-coagulation indicated that T-P removal was more stable than the biological method only. In addition, it was confirmed that T-N and organic materials as well as T-P were removed simultaneously by the electrochemical reaction in the bioreactor combined with electrolysis by more than the bioreactor only. PMID- 14753523 TI - Ten years experience of treating all flows from combined sewerage systems using package plant and constructed wetland combinations. AB - Severn Trent Water have been using constructed wetlands since 1991 as part of a flowsheet in which 6 DWF (Dry Weather Flow) is treated by rotating biological contactors (RBCs), and constructed reed beds used to provide both tertiary treatment and stormwater treatment. The company now has over 50 such sites using this flowsheet. Dimensioning and effluent quality achievable is reported and data presented showing the performance of reed beds treating stormwater alone or in admixture with RBC effluent. After 8-10 years of operation the condition of a number of storm reed beds was assessed to determine the remaining asset life. The majority seem likely to achieve at least another five years of operation before further review is needed. PMID- 14753524 TI - Greywater treatment by constructed wetlands in combination with TiO2-based photocatalytic oxidation for suburban and rural areas without sewer system. AB - In this study greywater treatment through constructed wetlands and subsequently through TiO2-based photocatalytic oxidation was investigated. Through constructed wetlands treatment the organic substances have been reduced greatly. For further removal of organic substances and pathogens, a TiO2-based photocatalytic oxidation process was used subsequently. The results showed that the treated greywater through constructed wetlands and subsequent through TiO2-based photocatalytic oxidation with short irradiation time (3 hours irradiation time) met the requirements of European bathing water quality easily. Therefore, the greywater treated with the processes combination can directly be reused for non potable purposes. Moreover, since residual organic substances through TiO2-based photocatalytic oxidation with long irradiation time can be eliminated almost totally, it is also possible that treated greywater is used for groundwater recharge as a drinking water resource. PMID- 14753525 TI - Cost-effective treatment solutions for rural areas: design and operation of a new package treatment plant for single households. AB - The design approach and operation of a newly developed package plant treating domestic sewage from single households were evaluated. Combining submerged aerated filter (SAF) technology with jet aeration and incorporating both into a compact and shallow tank resulted in a cost-effective treatment solution. A trial unit was permanently installed at a rural site, serving a single household. Jet aeration proved to be the best aeration method for the shallow bioreactor design. Further trials revealed a 50% reduction in suspended solids (SS) through the use of a static effluent filter and found that annual plant maintenance was vital to sustain stable operating conditions. Despite high variations in influent conditions, the trial unit produced good effluent quality during steady-state operation. Average effluent BOD5, COD and SS values were 19.6 mg l(-1), 98 mg l( 1) and 32 mg l(-1) achieving overall removal efficiencies of 94.2%, 85.9% and 87.6% respectively. However, effluent ammonia nitrogen (NH4-N) levels were found to be inconsistent varying from 9 mg l(-1) to over 60 mg l(-1). PMID- 14753526 TI - Design of water hyacinth ponds for removing algal particles from waste stabilization ponds. AB - In this study it was demonstrated that when water hyacinth ponds (WHPs) are used for polishing the effluent from waste stabilization ponds (WSPs), suspended solids (mostly algal particles) are efficiently separated, which also resulted in the reduction of insoluble forms of COD and nutrients. The high pH of the WSPs effluent was easily adjusted to 6-7 as it passed through the WHPs. However, the use of water hyacinth rapidly reduced dissolved oxygen at the first cell to less than three mg/L or very frequently to a level of anaerobic state. Reduction of suspended solids at the WHPs mainly depends on the detention time and pH. An empirical separation model incorporating the detention time and pH dependence was developed. PMID- 14753527 TI - Vermiculture as a tool for domestic wastewater management. AB - Organic waste management is a growing issue due to the unsustainable practices of its disposal. Sewage treatment plants are designed to treat wastewater to produce a safe effluent. However, one of the by-products, the sewage sludge which is disposed off in landfill or used as fertilizer in agricultural operation is high in pathogens. Sustainability can be achieved by Vermicomposting of organic matter which involves accelerated cycling of nutrients though a closed cycle whereby waste products are put to productive end use. Vermicomposting and vermifiltration are natural waste management processes relying on the use of worms to convert organic wastes to stable soil enriching compounds. Domestic wastewater management can be accommodated through these processes in a sustainable manner. A considerable reduction in pathogens has been noticed in the end product to a level that it can be safely applied to land. This paper provides an overview of the system characteristics of management systems utilising vermiculture, to manage wastewater. The process can be used in a small scale for household waste treatment to rural or urban waste management. PMID- 14753528 TI - Pre-treatment of domestic wastewater with pre-composting tanks: evaluation of existing systems. AB - A relatively new technology called pre-composting tank or Rottebehaelter, retaining solid material and draining water to a certain extent, has been found to be an interesting component of decentralised systems to replace the usual septic tank. Results of the investigation revealed that solid material which has been retained in the pre-composting tanks still contained a high percentage of water. However, there was no odour problem at and near the tanks. The pre composted materials have to be further composted together with household and garden wastes for a year prior to their use as soil conditioner. The filtrate is further treated in a constructed wetland. One of the major advantages of this system compared to other systems, such as septic tanks, is that it does not deprive agriculture of the valuable nutrients and soil conditioner from human excreta and does not require an expensive tanker truck. It can be the most appropriate system for application in regions where there is a demand for local reuse of the end product. It has to be stated that maintenance is a crucial factor. PMID- 14753529 TI - Oxidation processes and clogging in intermittent unsaturated infiltration. AB - Intermittent infiltration of wastewater through a non saturated sand bed is an extensive treatment process aimed at eliminating organic pollution, oxidizing ammonia and removing pathogens. A 1D numerical model, IPOX, has been worked out to simulate the transfer and oxidation of dissolved organic matter and nitrogen in unsaturated sand beds. IPOX was calibrated after real scale tests performed in Spain and Burkina Faso. Simulations allowed us to point out the influence of (i) kinetics on oxidation performances and (ii) biomass development on the process sustainability. These results brought a new light on the sizing and operation of infiltration percolation and soil aquifer treatment (SAT) plants. PMID- 14753530 TI - Intermittent sand filtration for wastewater treatment in rural areas of the Middle East--a pilot study. AB - This paper concentrates on Intermittent Sand Filtration (ISF) as a polishing stage for effluent from a facultative pond. During the three-year research program, the system operated with an influent flow-rate of 500-1,000 L/day and an average BOD concentration of 200-400 hydraulic and BOD loadings of 110-200 L/m2/day and 20-40 gBOD/m2/day, respectively. Flow to the ISF was applied intermittently with a different number of doses in each run. In addition, the effects of the frequency and the duration of rest periods (no feeding) were studied. Removal of 90-95% of BOD and 75-90% of COD and TSS was achieved consistently throughout the study period. Elevated levels of nitrification were observed with 95-100% removal of NH3. The ISF performed best when fed with 5-10 doses/day. Reducing the daily number of doses to 3/day at the same hydraulic loading rate resulted in a 20-30% reduction in removal efficiency. The 2-4 week rest period had no effect on the biological activity in the subsequent run. However, rest periods of more than 30 days were found to negatively affect removal efficiency. PMID- 14753531 TI - Sand filter clogging by septic tank effluent. AB - The aim of this study was to characterise conditions and factors affecting fine sand clogging by septic tank effluent on the basis of physical modelling. The physical model consisted of 12 sand columns dosed with sewage from one household (5 persons), preliminary treated in a septic tank. Hydraulic loadings of the sand filters were equal to 82 mm/d. The mean discharge from sand columns, measured as the effluent volume collected during 10 minutes, decreased significantly over the experiment period from 34 cm3/min in August 2000 to 20 cm3/min in August 2001 at the same temperature of about 20 degrees C. First the columns clogged almost completely after 480 days in December 2001, however six columns had remained unclogged till the end of the experiment (March 2002). The temperature had a significant impact on hydraulic conductivity. A vertical distribution of accumulated mass and biomass was investigated in partly clogged sand. Microscopic survey of the clogging layer showed a presence of live micro-organisms, residuals of dead micro-organisms, particularly pieces of small animal armour and many fibres. These particles accelerated the accumulation of solids in the upper clogging layer. The study indicated that temperature impact on the filter hydraulic conductivity was more significant for biological activity, than for sewage viscosity. PMID- 14753532 TI - A small scale hydroponics wastewater treatment system under Swedish conditions. AB - A treatment plant using conventional biological treatment combined with hydroponics and microalgae is constructed in a greenhouse in the area of Stockholm, Sweden. The treatment plant is built for research purposes and presently treats 0.559 m3 of domestic wastewater from the surrounding area per day. The system uses anoxic pre-denitrification followed by aerobic tanks for nitrification and plant growth. A microalgal step further reduces phosphorus, and a final sand filter polishes the water. During a three week period in July 2002 the treatment capacity of this system was evaluated with respect to removal of organic matter, phosphorus and nitrogen. 90% COD removal was obtained early in the system. Nitrification and denitrification was well established with total nitrogen reduction of 72%. Phosphorus was removed by 47% in the process. However, higher phosphorus removal values are expected as the microalgal step will be further developed. The results show that acceptable treatment can be achieved using this kind of system. Further optimisation of the system will lead to clean water as well as valuable plants to be harvested from the nutrient rich wastewater. PMID- 14753533 TI - Treatment of mountain refuge wastewater by fixed and moving bed biofilm systems. AB - Tourists visiting mountain refuges in the Alps have increased significantly in the last decade and the number of refuges and huts at high altitude too. In this research the results of an intensive monitoring of a wastewater treatment plant (WWTP) for a tourist mountain refuge located at 2,981 m a.s.l. are described. Two biofilm reactors were adopted: (a) a Moving Bed Biofilm Reactor (MBBR); (b) a submerged Fixed Bed Biofilm Reactor (FBBR). The aims of this research were: (i) the evaluation of the main parameters characterising the processes and involved in the design of the wastewater plants, in order to compare advantages and disadvantages of the two tested alternatives; (ii) the acquisition of an adequate knowledge of the problems connected with the wastewater treatment in alpine refuges. The main results have been: (i) a quick start-up of the biological reactors obtainable thanks to a pre-colonization before the transportation of the plastic carriers to the refuge at the beginning of the tourist season; (ii) low volume and area requirement; (iii) significantly higher removal efficiency compared to other fixed biomass systems, such as trickling filters, but the energy consumption is higher. PMID- 14753534 TI - Removal of ammonia from tannery effluents in a reed bed constructed wetland. AB - A horizontal subsurface flow reed bed, Pragmites australis, constructed wetland system was operated continuously for one year at 5 different hydraulic retention times between 5 days-11 days with real tannery industry effluent to investigate mainly NH4-N and COD removal performance of the system. The bed was also operated at different initial NH4-N concentrations between 10-30 mg/L by adding NH4Cl into the tannery effluent. The results indicated that ammonia-nitrogen removal is significantly affected by hydraulic retention time while COD is not. The optimum HRT was determined as 8 days with over 95% NH4-N and around 30% COD removal efficiency. The system shows higher removal performance at high initial NH4-N concentrations. Almost complete (99%) NH4-N removal and over 40% COD removal was obtained at (NH4-N)0 = 20 mg/L and HRT = 7 days. PO4-P and total chromium removal were not significantly affected by operating conditions PMID- 14753535 TI - Performance of a small wastewater stabilisation pond system in tropical climate in Dar es Salaam, Tanzania. AB - Waste stabilisation ponds (WSP) are widely used in Tanzania. Their traditional design focuses on BOD and pathogen removal, but nutrient removal is equally important. WSP performance was evaluated to come up with information that would be used to evaluate pond performance. Samples were analysed twice a month for six months. Results showed total and faecal coliform removal by 4 log units (99.96 and 99.98% respectively). There was partial reduction of COD, BOD, (46% and 27% respectively), conductivity and total dissolved solids (32.6 and 32.4% respectively). Variation for the nutrients like inorganic phosphorus, ammonia nitrogen, nitrite nitrogen, and nitrate nitrogen was not consistent and there was practically no reduction, although there is some nitrification taking place. From this study it may be concluded that BOD and nutrients may not be useful to evaluate pond performance. Instead parameters such as conductivity, total dissolved solids, coliform bacteria, dissolved oxygen, pH and chlorophyll would be suitable for that purpose, due to their consistent variation within cells of the pond system. But BOD and nutrient removal are important and have to be improved to enhance treatment in the WSP. PMID- 14753536 TI - Low cost anaerobic system for Indonesia: single baffled septic tank. AB - The insertion of a single baffle into a laboratory septic tank to mix incoming feed with sludge has been shown to improve anaerobic degradation of the feed. This is particularly true of soluble organic matter such as glucose. Oil or cellulose fed separately does not undergo degradation. It is expected however that a balanced feed such as sewage will be better degraded. PMID- 14753537 TI - Treatment of domestic sewage at low temperature in a two-anaerobic step system followed by a trickling filter. AB - The treatment of domestic sewage at low temperature was studied in a two anaerobic-step system followed by an aerobic step, consisting of an anaerobic filter (AF) + an anaerobic hybrid (AH) + polyurethane-foam trickling filter (PTF). The AF+AH system was operated at a hydraulic retention time (HRT) of 3+6 h at a controlled temperature of 13 degrees C, while the PTF was operated without wastewater recirculation at different hydraulic loading rates (HLR) of 41, 15.4 and 2.6 m3/m2/d at ambient temperature (ca. 15-18 degrees C). The AF reactor removed the major part of the total and suspended COD, viz. 46 and 58% respectively. The AH reactor with granular sludge was efficient in the removal and conversion of the anaerobically biodegradable COD. The AF+AH system removed 63% of total COD and converted 46% of the influent total COD to methane. At a HLR of 41 m3/m2/d, the COD removal was limited in the PTF, while at HLR of 15.4 and 2.6 m3/m2/d, a high total COD removal of 54-57% was achieved without a significant difference between the two HLRs. The PTF was mainly efficient in the removal of particles (suspended and colloidal COD removal were 75-90% and 75-83% respectively), which were not removed in the anaerobic two-step. The overall total COD removal in the AF+AH+PTF system was 85%. Decreasing the HLR from 15.4 to 2.6 m3/m2/d, only increased the nitrification rate efficiency in the PTF from 22% to 60%. Also, at HLR of 15.4 and 2.6 m3/m2/d, PTF showed a similar removal for E. coli by about 2 log. Therefore, the effluent of AF+AH+PTF system can be utilised for restricted irrigation in order to close water and nutrients cycles. Moreover, such a system represents a high-load and a low-cost technology, which is a suitable solution for developing countries. PMID- 14753538 TI - Effect of high salinity on anaerobic treatment of low strength effluents. AB - In anaerobic treatment, it is obligatory to know the effect of potentially inhibitory compounds due to the fact that methane formation may retard severely and may proceed slower than organic acid production. One of the most important inhibitory substances in anaerobic treatment is high salinity. In many cases, the main collectors of a municipal sewer system should have been built in the coastal zone and below the ground water level due to the available topography of wastewater catchments area, which is carrying the risk of seawater infiltration. Besides, one of the most convenient methods for leachate control is to treat landfill leachates with domestic wastewaters in the central municipal wastewater treatment plants such as in Istanbul. Thus, the nitrogen load of the treatment plants increase significantly. In this study, the effects of high salinity and ammonium nitrogen levels on mesophilic anaerobic tretament processes were investigated. In the first part of the study, high salinity effect on anaerobic treatment was investigated by feeding synthetic wastewater containing high salinity between 0.15%-1.5% ratios. In the second part of the study, the simultaneous effect of high salinity and ammonia (1.5% salinity+1,000 mg NH3/l) was examined by a lab-scale Upflow Anaerobic Sludge Bed Reactor (UASBR). Results indicated no significant inhibition in both cases and effective COD removals (89%) and total biogas productions having methane content of 84% could be achieved. PMID- 14753539 TI - Small sewage treatment system with an anaerobic-anoxic-aerobic combined biofilter. AB - The objective of this study was to investigate a small sewage treatment system that could improve nitrogen and BOD5 removal efficiency as well as generate less solid using an anaerobic-anoxic-aerobic biofiltration system. Wastewater temperature was in the range of 14-25 degrees C, and hydraulic residual times were 12 h for each reactor. The upflow anaerobic digester equipped with anoxic filter was fed with both raw sewage and recycled effluent from the aerobic filter to induce denitrification and solid reduction simultaneously. In the subsequent aerobic filter, residual organic carbon and ammonia might be oxidized and finally nitrate formed. In the anaerobic reactor, about 71% of influent TCOD was removed by sedimentation of the un-filterable COD at the recycle ratio of 300%. Another 20% of influent TCOD was removed in the anoxic filter by denitrification of the recycled nitrate. After 100 days operation, solid reduction and nitrification efficiency were about 30% and 95%, respectively. Overall removal efficiencies of COD and total nitrogen (T-N) were above 94% and 70% at the recycle ratio of 300%, respectively. Total wasted solid from the system after 100 days operation was about 316 g, which was only 44% of the solid generated from a controlled activated sludge system operated at sludge retention time of 8 days. PMID- 14753540 TI - The UASB reactor as an alternative for the septic tank for on-site sewage treatment. AB - Although septic tanks are amply used for on site sewage treatment, these units have serious drawbacks: the removal efficiency of organic material and suspended solids is low, the units are costly and occupy a large area and operational cost is high due to the need for periodic desludging. In this paper an innovative variant of the UASB reactor is proposed as an alternative for the septic tank. This alternative has several important advantages in comparison with the conventional septic tank: (1) Although the volume of the UASB reactor was about 4 times smaller than the septic tank, its effluent quality was superior, even though small sludge particles were present, (2) desludging of the UASB reactor is unnecessary and even counterproductive, as the sludge mass guarantees proper performance, (3) the UASB reactor is easily transportable (compact and light) and therefore can be produced in series, strongly reducing construction costs and (4) since the concentration of colloids in the UASB effluent is much smaller than in the ST effluent, it is expected that the infiltration of the effluent will be much less problematic. PMID- 14753541 TI - Degradation of xenobiotic substances using sulfate-reducing bacteria in a UASB reactor. AB - An upflow anaerobic sludge blanket (UASB) reactor was successfully applied to continuous degradation of ferric ethylene diamine tetraacetate (Fe-EDTA) as a typical xenobiotic substance contained in photo-processing wastewater. The sludge in the UASB reactor had an abundance of sulfate-reducing bacteria (SRB), which had been anaerobically cultivated in a sulfate-rich culture medium including Fe EDTA and yeast-extract as the carbon sources. Since the prominent reductions of Fe-EDTA and sulfate ion were observed, the contribution of SRB to Fe-EDTA degradation in the UASB reactor was confirmed. The aggregated sludge in the UASB reactor became gradually large reaching steady state with an equivalent diameter of 60-90 microm after 124 days operation. An increase of the amount of yeast extract addition to feed solution improved the Fe-EDTA removal efficiency up to 90%. Moreover, the combination of ozone treatment with SRB treatment further improved removal efficiency of total organic carbon (TOC) in an actual photo processing wastewater composed of fixing and developing wastes. PMID- 14753542 TI - Experimental study on carbon removal in biological aerated filters. AB - The aim of the present work was to evaluate the performance of a pilot-scale BAF in terms of removal of organic matter and suspended solids to obtain a highly polished effluent. The first part of the research was the evaluation of the optimal filter media for a full scale BAF. Mechanical and biological tests were performed over four materials: glass, plastic, pozzolan and expanded clay (Arlita) and the results obtained showed that the plastic spheres and the Arlita particles were the optimal materials for both the mechanical and biological requirements. Hence, a down-flow pilot scale BAF was set up in the laboratory to treat a synthetic medium. As filter media first plastic spheres and then Arlita spheres were used. Carbon removal studies were carried out at several influent COD concentrations, specific removal efficiency and COD profiles along the height of the filter were determined and used to analyze the process. Validation and calibration of a mathematical model formulated for carbon removal, were also performed by using the experimental data obtained. The results showed that this system allows us to achieve the more strict limits on final effluent. PMID- 14753543 TI - A new low-cost biofilm carrier for the treatment of municipal wastewater in a moving bed reactor. AB - The Moving Bed Biofilm Reactor has proven to be an efficient system in wastewater treatment and has become a viable solution for small treatment plants. The main objective of this research was to analyse the performance of a moving bed reactor using low-cost local material when fed with municipal wastewater. A pilot reactor with a total volume of 900 litres was built and it was fed continuously with municipal wastewater. The operation of the system was adjusted to six different organic loading rates. The biofilm carrier was polyethylene tubing with internal diameter of 1.1 cm, cut into pieces of 1.2 cm. The tested material offered a specific surface area of 590 m2/m3. Air was provided with a fine-bubble diffuser. The main results show that the reactor performance was stable and predictable. The COD removal confidently behaves according to a general hyperbolic kinetic equation. The maximal total COD removal attained was 81%. Nitrification was observed only for organic loads with values under 5.7 gCOD/m2 x d. Good adherence of the microorganisms was observed for the applied organic loading rates. After several months of operation, the material showed no signs of abrasion or deformation. The sludge production behaved linearly with the organic load reaching 979 gTSS/d with the highest organic load of 35.7 gCOD/m2 x d. The amount of microorganisms attached to the carrier increased with the organic load tending to an asymptotical maximal value of 17.3 g/m2 (as dry solids). Mean cellular retention times from 2.0 to 23.1 days were determined. PMID- 14753544 TI - Moving bed biofilm reactors: a small-scale treatment solution. AB - The design and operational suitability of the moving bed process for small-scale sewage treatment systems were evaluated. A prototype plant was installed at a new housing development near Winchester, UK, and operated under different conditions over an eight-month period. During normal operation, the MBBR plant produced a good quality effluent with average values for BOD5, COD, SS and NH4-N of 15.6 mg l(-1), 65.9 mg l(-1), 21 mg l(-1) and 4.7 mg l(-1), respectively. The plant was further subjected to organic and hydraulic shock loads. The treatment performance remained high over the organic shock period, however, hydraulic surges of up to 10 times the design flow caused a loss of solids from the final settlement stage. While organic carbon removal resumed to over 80% within a single week after a prolonged power failure, effluent NH4-N values remained high for a period of three weeks. Besides producing a good quality effluent, the MBBR plant proved to be simple and reliable to operate, thus providing a viable treatment solution for small-scale applications. PMID- 14753545 TI - A software nitrate sensor based on ammonium and redox signals. AB - We have computed the nitrate concentration in the activated sludge in real-time using a model, which is a subset of ASM1. The model is in operation at two WWTPs where oxygen, ammonium and redox are measured online in the aeration tanks. The model uses these measurements to continuously adjust its values for the influent ammonium concentration, the nitrification rate, the denitrification rate and the net hydrolysis. Then it computes the nitrate concentration. This value is updated every 10 s. The model results have been compared with the output from a Dr Lange in-situ nitrate sensor at one of these WWTPs. The systematic difference between these two measurements is less than 0.2 mgN L(-1) and 90% of the differences are between -1.1 and 1.1 mgN L(-1). PMID- 14753546 TI - An off-line respirometric procedure to determine inhibition and toxicity of biodegradable compounds in biomass from an industrial WWTP. AB - Determining whether a certain compound is toxic (or inhibitor) or not to a biomass of a WWTP is crucial in order to avoid WWTP malfunction. Respirometric techniques have been commonly used to assess the toxicity of a certain compound by evaluating the oxygen uptake rate (OUR) profile obtained when a pulse of substrate is added to endogenous biomass and comparing it with the OUR profile obtained when the pulse is a mixture of substrate and the possible toxic or inhibitor (PTI). However, when using this method with biodegradable compounds some difficulties arise because the PTI consumption implies oxygen consumption as well. In this study, a modified procedure for toxicity assessment using respirometry is developed which overcomes the difficulties caused by the toxic biodegradability. This procedure is based on the comparison of different respirograms obtained with pulses of control substrate before and after adding the PTI and with a pulse of PTI and control substrate together. Moreover, some examples are shown with p-phenylenediamine and phenol as an example of toxic and inhibitor for nitrifying biomass. PMID- 14753547 TI - Treatment of landfill leachate--high tech or low tech? A case study. AB - At the sanitary landfill of the city of Penzberg (Germany), two diverse approaches to leachate treatment were studied as parts of a three-stage treatment concept. The performance of a simple aerobic pond was compared to that of an advanced multistage treatment unit, the latter comprising a membrane biological reactor and a two-stage activated carbon filter. For 274 days of the year (75%) the pond was able to provide sufficient treatment even under cold weather conditions. For temperatures lower than 5 degrees C, a higher biomass content and temporal storage of the raw leachate (e.g. increasing hydraulic retention time) could close the gap of insufficient treatment. In contrast, the advanced treatment system could only accomplish limited treatment capabilities due to insufficient maintenance, low loading conditions and deficient coordination between the individual treatment steps. As a result, degradation rates were low and operational problems frequent. Limits for Ntot were exceeded regularly (Ntot,e = 60-70 mg/L), throughput broke down and excessive nitrite production occurred (NO2-Ne = 10 mg/L) as a result of microbial activity inside the activated carbon filters. This case study clearly suggests aerobic ponds as an appropriate solution for the treatment of landfill leachate in areas where operational independence is essential. PMID- 14753548 TI - Design procedure for carbon removal in contact stabilization activated sludge process. AB - The contact stabilization activated sludge process (CSASP) has been adopted and applied in full scale plants treating domestic sewage. Acquiring smaller volume thereby reducing costs and achieving the same treatment efficiency as the conventional activated sludge plants are the main advantages of the CSASP. This activated sludge modification is especially ideal for small to mid-scale plants where influent wastewater contains a high fraction of particulate COD. The simulation results reveal that the same amount of mass sludge can be retained in the system with almost 30% volume reduction compared to conventional activated sludge plants. It is also found that the CSASP treatment efficiency increases when the particulate COD fraction of an influent domestic wastewater increases. The most important process component in the design of CSASP is the sludge distribution factor (alpha) which directly affects the effluent characterization. Having most of the biomass in the stabilization reactor (meaning low alpha values) decreases the nominal hydraulic retention time (thetaHN) of the system. However the sludge distribution factor must be high enough to ensure an acceptable effluent quality. PMID- 14753549 TI - Investigation of sludge re-circulating clarifiers design and optimization through numerical simulation. AB - In steam thermal power plants (TPP) with open re-circulating wet cooling towers, elimination of water hardness and suspended solids (SS) is performed in clarifiers. Most of these clarifiers are of high efficiency sludge re-circulating type (SRC) with capacity between 500-1,500 m3/hr. Improper design and/or mal operation of clarifiers in TPPs results in working conditions below design capacity or production of soft water with improper quality (hardness and S.S.). This causes accumulation of deposits in heat exchangers, condenser tubes, cooling and service water pipes and boiler tubes as well as increasing the ionic load of water at the demineralizing system inlet. It also increases the amount of chemical consumptions and produces more liquid and solid waste. In this regard, a software program for optimal design and simulation of SRCs has been developed. Then design parameters of existing SRCs in four TPPs in Iran were used as inputs to developed software program and resulting technical specifications were compared with existing ones. In some cases improper design was the main cause of poor outlet water quality. In order to achieve proper efficiency, further investigations were made to obtain control parameters as well as design parameters for both mal-designed and/or mal-operated SRCs. PMID- 14753550 TI - Experimental study on sequencing batch biofilm reactor with biological filtration (SBBR-BF) for wastewater treatment. AB - A novel wastewater treatment technology combining a sequencing batch biofilm reactor and biological filtration in an SBBR-BF system was presented. Elastic plastic filaments were fixed as biofilms carrying media. Particle materials (sand or anthracite) and the settled sludge constituted the filtration layer. In the laboratory studies, operating results of SBR, SBBR and SBBR-BF were compared. Better quality and stable water quality of effluent could be achieved in SBBR-BF because the fixed film and filtration layer were added in the reactor. Other laboratory experiment results indicated that slow filtration, cycle water stirring and backwashing making use of the settled supernatant are successful methods for preventing clogging and saving energy. The velocity and headloss of filtration were significantly impacted by different MLSS concentration. The MLSS concentration in the reactor must be less than 1,400 mg/L for optimal results. The average velocity of filtration ranging from 0.6 to 1.0 m/h, the backwash velocity of 10-15 m/h and the backwash time of 20 seconds are recommended according to the laboratory experiment. On-site experiment and study showed that SBBR-BF is a stable and efficient system for domestic wastewater treatment, and is particularly suited for small wastewater treatment plants, because of the simple operation and compact installation. PMID- 14753551 TI - Performance of a hybrid SBR with fixed bed and suspended growth. AB - A hybrid SBR system combined with fixed bed (media) and suspended growth zones was developed to improve both nitrogen and phosphorus removal. About 27% of the tank volume was filled with clay media to make a fixed bed in this system, and additional air was applied under the bed to wash the microbes from the media during the oxic stage to prevent the bed from clogging. This hybrid SBR system could eliminate the backwashing requirement for SBBR (sequencing batch biofilm reactor). This system showed a stable nitrification even at low temperature and shock load conditions. The specific reaction rates indicated the fixed bed zone had higher microbial activities for nitrification and denitrification, while the suspended growth zone had higher microbial activities for phosphorus release and uptake. In addition, the use of external sludge storage also increased both phosphorus and nitrogen removal. The effluent COD, nitrogen and phosphorus concentrations were respectively less than 15, 10.5 and 0.6 mg/L with weak sewage of 230 mg/L COD, 35 mg/L TKN and 5.3 mg/L TP. PMID- 14753552 TI - Effect of temperature on the nitrogen removal performance of a sequencing batch reactor treating tannery wastewater. AB - The effect of temperature on the nitrogen removal performance of the sequencing batch reactor technology is evaluated for tannery wastewater. The study involved the operation of a pilot-scale sequencing batch reactor installed on site to treat the plain-settled effluent. The nitrogen balance of the system is observed for a wide temperature range between 9 to 30 degrees C. The results are evaluated by means of model calibration of COD, nitrate and ammonia nitrogen concentration profiles during cyclic operation. The fates of the major nitrogen parameters are also interpreted on the basis of fundamental stoichiometry for nitrification and denitrification. PMID- 14753553 TI - Feasibility study of sequencing batch reactor system for upgrading wastewater treatment in Malaysia. AB - The objective of this study was to assess the feasibility of the Sequencing Batch Reactor (SBR) system for implementation in Malaysia. Theoretical, field, laboratory investigations, and modelling simulations have been carried out. The results of the study indicated that the SBR system was robust, relatively cost effective, and efficient under Malaysian conditions. However, the SBR system requires highly skilled operators and continuous monitoring. This paper also attempted to identify operating conditions for the SBR system, which optimise both the removal efficiencies and the removal rates. The removal efficiencies could reach 90-96% for COD, up to 92% for TN, and 95% for SS. An approach to estimate a full operational cycle time, to estimate the de-sludging rate, and to control the biomass in the sludge has also been developed. About 4 hours react time was obtained, as 2.25 hours of nitrification with aerated slow fill and 1.75 hour of denitrification with HAc addition as an additional carbon source. Inefficient settling was one of the problems that affect the SBR effluent quality. The settling time was one hour for achieving Standard B (effluent quality) and 2 hours for Standard A. PMID- 14753554 TI - An alternate oxic-anoxic process automatically controlled. Theory and practice in a real treatment plant network. AB - A simple mathematical model of an alternate oxic-anoxic process has been elaborated. It enables us to optimise the cycle time on the basis of maximum nitrates concentration in the effluent and the desired nitrogen removal performance. At the same time the model can be employed to verify the impact of the variations of flow rate and influent characteristics as well as the operational parameters of the process. Actually, the model confirms the process efficiency but its feasibility in real plants needs a local or remote process control. To verify these theoretical conclusions a real wastewater plant (700 PE) has been upgraded in an alternate oxic-anoxic process. It was implemented with software able to elaborate the data of dissolved oxygen concentration and oxidation reduction potential. Moreover, the evaluation of the flexing points was performed to manage mixer and blowers. A one-year experience of plant management allowed us to obtain very high nitrogen removal. However, the performances were different during wet or dry weather periods. The statistical analysis of probe signals evaluation confirmed the capability of the control device to detect the flexing points during the anoxic phase (70-94%). On the other hand, the capability of detecting the DO signal was lower, in particular when the oxygen demand was similar to the amount of supplied oxygen. The hourly variations of flow rate and mass loading determines different conditions for starting the anoxic phase: over aeration, over loading and the equivalence of oxygen demand and supply, are the main factors determining the blowers stopping. PMID- 14753555 TI - Integrated control of nitrate recirculation and external carbon addition in a predenitrification system. AB - The integrated control of nitrate recirculation and external carbon addition in a predenitrification biological wastewater treatment system is studied. The proposed control structure consists of four feedback control loops, which manipulate the nitrate recirculation and the carbon dosage flows in a highly coordinated manner such that the consumption of external carbon is minimised while the nitrate discharge limits (based on both grab and composite samples) are met. The control system requires the measurement of the nitrate concentrations at the end of both the anoxic and the aerobic zones. Distinct from ordinary control systems, which typically minimise the variation in the controlled variables, the proposed control system essentially maximises the diurnal variation of the effluent nitrate concentration and through this maximises the use of influent COD for denitrification, thus minimising the requirement for external carbon source. Simulation studies using a commonly accepted simulation benchmark show that the controlled system consistently achieves the designated effluent quality with minimum costs. PMID- 14753556 TI - Nitrogen removal during secondary treatment by aquatic systems. AB - Within the context of this study, two lab-scale aquatic plant reactors consisting of duckweed (Lemna minor) ponds, were investigated for the removal of nitrogen forms during the secondary treatment of domestic wastewater. TKN, NH3-N and NO3-N parameters have been measured in both reactors for hydraulic retention times ranging from 3.3 days to 23 days and at various distances from the inlet of reactors. The results were evaluated for hydraulic retention times, hydraulic loading rates and mass loading rates. I was concluded that hydraulic and mass loading parameters were more meaningful than hydraulic retention time. Optimum nitrogen removal values of hydraulic loading rate and mass loading rate were found to be 1.2 cm/day and 90-160 mg TKN/m2-day, respectively. At the higher and lower loading rates, nitrogen removal efficiency was lower than those at optimum conditions. Effluent TKN concentration was around 2.5 to 3.0 mg/l while NH3-N concentration was almost zero at these loading conditions. On the other hand, effluent NO3-N concentrations changed between 7 mg/l to 11 mg/l. When investigating the longitudinal profile, values were reduced rapidly along the reactors. It was concluded that most of the nitrogen conversion occurred at the beginning of the reactor. PMID- 14753557 TI - Effects of SRT and DO on N2O reductase activity in an anoxic-oxic activated sludge system. AB - Nitrous oxide (N2O) is emitted from wastewater treatment processes, and is known to be a green house gas contributing to global warming. It is thus important to develop technology that can suppress N2O emission. The effects of sludge retention time (SRT) and dissolved oxygen (DO) on N2O emission in an anoxic-oxic activated sludge system were estimated. Moreover, the microbial community structure in the sludge, which plays an important role in N2O suppression, was clarified based on nitrous oxide reductase (nosZ) gene analysis by molecular biological techniques. The results showed that under low SRT conditions, nitrification efficiency was reduced and the N2O emission rate in the oxic reactors was increased. It was also observed that N2O emission was enhanced under low DO conditions, where the available oxygen is insufficient for nitrification. Moreover, molecular analysis revealed that the clones identified in this study were closely related to Ralstonia eutropha and Paracoccus denitrificans. The fact that the identified sequences are not closely related to known culturable denitrifier nosZ sequences indicates a substantial in situ diversity of denitrifiers contributing to N2O suppression, which are not reflected in the cultivatable fraction of the population. The further application of these new molecular techniques should serve to enhance our knowledge of the microbial community of denitrifying bacteria contributing to N2O suppression in wastewater treatment systems. PMID- 14753558 TI - Comparison of intermittently aerated continuous and batch biofilm reactor in nutrient removal. AB - Removal efficiency of TOC ranged between 86-89% in an intermittently aerated reactor. High efficiency in TKN removal and nitrification was found at lower applied load or longer retention time such as 2 days. TKN removal and nitrification efficiency was found to be 17-96% and 35-99% respectively. Through examination and comparison of the removal efficiencies, the stability of nitrification/denitrification and the biological phosphorus removal it was found that the sequencing batch feeding system gave a higher performance in total nitrogen and phosphorus removals. In the SBR reactor, nitrogen removal efficiency was mainly controlled by organic loading. Nitrification efficiency ranged between 31-56%. Nearly complete denitrification was observed in the sequencing batch reactor. PMID- 14753559 TI - Nutrient removal of ammonia rich effluents in a sequencing batch reactor. AB - The aim of this study was to develop an appropriate operating strategy for ammonia removal of young landfill leachate in a lab-scale sequencing batch reactor, SBR. SBR was operated at five different phases by changing the aerobic cycle time and external carbon source during the denitrification process. SBR provides the opportunity to arrange the operating periods according to variable conditions such as wastewater characterization in order to optimise the performance of the system. By monitoring the variations occurring in each period during a full cycle an appropriate operating strategy may be defined. The main problem faced during the experimentation period particularly was due to use of raw wastewater with high NH4-N content as an external carbon source, as it affected denitrification performance to a great extent. This trouble was overcome if calcium acetate was used as the external carbon source instead of the raw wastewater. In case of using a suitable aeration period and the convenient external carbon source, high ammonia removals were observed. PMID- 14753560 TI - Consumer managed co-operative--a solution for progressing wastewater management in rural areas. AB - In Finland the question of appropriate wastewater treatment in rural areas has become very prominent during the last years. The new Environmental Protection Act stipulates that wastewaters must be treated to the extent that they cannot have a negative impact on nature. The Ministry of the Environment is currently preparing a decree specifying the requirements. The draft of the decree proposes that on site treatment units should decrease the BOD load by 90, total phosphorus load by 85 and total nitrogen load by 40 per cent. To meet the new requirements, the old systems that include septic tanks only need more efficient wastewater treatment methods. Whatever technical solution is selected, the house owner must pay for it. At the moment, even the septic tanks are emptied and maintained irregularly. More sophisticated wastewater treatment methods definitely need more maintenance, which cannot be made the sole duty of the house owners. One potential organisational alternative for managing wastewater treatment in rural areas is the co-operative. Finland has one such pioneering co-operative formed for on-site sanitation. Varsinais-Suomi Water Services Co-operative provides the house owner with professional assistance in wastewater treatment at a reasonable cost. Suvisaari Water Services Co-operative is another new organisation selling sewerage services to its members. But its technology is different: this co operative operates an LPS-sewerage system instead of on-site treatment. PMID- 14753561 TI - Development of a knowledge-based decision support system for identifying adequate wastewater treatment for small communities. AB - The identification of adequate treatment for small communities is a complex problem since it makes it necessary to combine aspects of the community and landscape, the receiving environment, and the available wastewater treatment technologies. This paper presents the development and implementation of a Knowledge-Based Decision Support System (KB-DSS) to tackle this problem. Different knowledge sources have been consulted in order to make up a comprehensive and accurate knowledge base. The core of the KB-DSS embraces two objectives. The first one is to assist in the selection of the treatment level adequate to fulfil the target quality standards for the receiving environment. The second one is to select the specific type of treatment. The KB-DSS is being applied to each one of the 3,482 different small communities comprised in the Small Communities Wastewater Treatment Plan of Catalonia, grouped according to river catchments. This paper also summarizes the different steps involved in the operation of the knowledge-based DSS when solving a real case study. PMID- 14753562 TI - A system for on-site performance evaluation of small wastewater treatment plants according to the European procedure. AB - The new European standard project for performance evaluation of on-site systems (prEN 12566-3) provides a test procedure at user sites for individual on-site wastewater treatment. For each system, two plants of the same size have to be tested, one operating at 50% and the second one with a load greater than 75% of the nominal capacity. The test duration is one year and several stress tests are included in the program (holidays, bath water discharge and power breakdown). Flow based composite 24 h samples, on the influent and the effluent have to be used, with 30 day intervals for influent and 15 days for effluent. On each sample, the following measurements have to be done: BOD5 or COD, suspended solids, temperature, power consumption, daily flow. It appears that very few systems have been reported in the literature to follow up such facilities, especially describing how to sample at the inlet of on-site individual equipment. In order to obtain representative samples at the inlet and at the outlet of those on-site treatment systems, we have designed an original mobile sampler system. The whole system ensures the flow measurement and flow based sampling as well. In this paper, we present the different parts of the sampling system (pump, flowmeters, ...), its validation and the results obtained at 5 user sites during the first 9 month period. Preliminary results are very interesting because they clearly demonstrate the need for an efficient on-site control of those user sites and for better legislation in this domain. PMID- 14753563 TI - Carbonate addition--an effective remedy against poor activated sludge settling properties and alkalinity conditions in small wastewater treatment plants. AB - The impact of marble powder on the performance of small seasonal treatment plants dealing with high ammonia concentrated wastewater has been investigated. The carbonate has been added to one of two parallel lab-scale SBR-systems as an alkalinity depot and a bio-carrier. The focus of the monitoring program was put on the measurement of carbon fractions in the liquid and the solid phase and on the alkalinity conditions. Monitoring results show a significant correlation between carbonate dosage (and corresponding inorganic carbon concentration and pH value) and sludge volume index and nitrification rate. PMID- 14753564 TI - Improving nitrogen removal in predenitrification-nitrification biofilters. AB - The effect on NH4-N removal rates in nitrification biofilters of filtered biodegradable COD and particulate COD leaving predenitrification biofilters was studied in a lab scale plant configured with the separated system of biofilters for secondary nitrogen removal from urban wastewaters. Applying a typical COD load of 11 kg/m3 x day to the predenitrification biofilter and maximizing its COD removal by adding nitrates or by operating an improved control of the internal recycle, only 60% removal of filtered biodegradable COD was found. This value corresponds to the complete removal of the readily biodegradable substrate (30% of influent filtered COD) and 36% of filtered slowly biodegradable substrate (50% of influent COD). The remaining 64% of the latter entered the nitrification biofilter, causing competition between heterotrophs and nitrifiers for dissolved oxygen in the inner layers of the biofilm. Consequently the nitrification rate had relatively low values (0.5 kgN/m3 x d) at 14 degrees C despite using dissolved oxygen levels of 6 mg/l. This behaviour may explain the lower nitrification rates obtained in some cases of nitrification biofilters compared to those in tertiary nitrification after activated sludge processes. The particulate COD entering the nitrification biofilter is associated with the suspended solids leaving the denitrification biofilter which are adsorbed by the external layers of the biofilm, increasing its thickness. The activity of the nitrifiers was affected because of a lack of oxygen when the thickness was left to grow considerably. Therefore no significant particulate COD effect is expected to occur as long as backwashing is carried out with the appropriate frequency. PMID- 14753565 TI - Optimizing nitrogen removal in the BioDenitro process. AB - The potential and limits of different configurations of the BioDenitro alternating process to suit the various design cases that can arise depending on the wastewater characteristics, space necessary and effluent nitrogen requirements were analysed through simulations of the activated sludge model No. 2. The first analysis involved the combination in one cycle of the main phase in the alternating reactors with an aerated phase having the two reactors in aerobic conditions and/or a hydraulic phase using the flow only in the anoxic reactor. This option has been found to have a very high potential for cases with strict requirements concerning effluent total nitrogen, but limited when the requirements are low effluent NH4-N, relatively high effluent total nitrogen and minimum solids and hydraulic retention times. When the latter conditions have to be fulfilled the incorporation of a post-aeration reactor to the alternating reactor was found to be very effective. In addition the configuration is very flexible because multiple combinations of post-aeration reactor volumes and in the duration of the different phases in the alternating reactors can be selected to achieve effluent nitrogen requirements. This flexibility is limited to the use of moderate values in the post-aeration reactor volumes and in the duration of the aerated phase. An experimental trial of the latter configuration was carried out and demonstrated its operational simplicity by achieving the desired nitrogen requirements in the effluent simply by changing the duration of the aerated phase for a given post-aeration reactor volume. From the experimental results an enhanced simultaneous nitrification-denitrification at the start of aeration in the alternating reactors was found and the ASM2 model was shown to have a satisfactory predictive capacity. PMID- 14753566 TI - Upgrading of the biotreatment in a pharmaceutical industry with activated carbon addition. AB - The impact of powdered activated carbon (PAC) on the biotreatment of a chemical synthesis wastewater emerging from a pharmaceutical industry was studied. The aim of this addition was to decrease inhibitory and non-biodegradable organics. In the first step, PAC was directly added to activated sludge. In the next step, the wastewater was first contacted with PAC and then treated in an activated sludge reactor. COD, oxygen uptake rate (OUR) and ultraviolet-visible (UV-Vis) spectral measurements were carried out for the evaluation of performance. Direct PAC addition and PAC pretreatment led to similar results, but direct PAC addition was considered more practical than pretreatment with PAC. With the addition of PAC, the level of non-biodegradable matter and the colour of the wastewater were significantly reduced. In any case, the addition of PAC increased the OUR of activated sludge, indicating that inhibition could be decreased. The combined evaluation of OUR, spectral parameters and COD gave a better insight into the processes taking place than the COD parameter alone. PMID- 14753567 TI - Enhancing biological nitrogen removal in a small wastewater treatment plant by regulating the air supply. AB - Most of the small wastewater treatment plants in Catalonia were not designed to remove nutrients. However, the large safety margins built into their original engineering design has meant that, in many cases, it has been possible to remove part of the nitrogen without being controlled. This paper focuses on the practical experience carried out in one of these facilities, the Bisbal wastewater treatment plant, aimed at enhancing the biological nitrogen removal using simple, low-cost, easy-to-operate measurement and control. The control strategy, which is based on air supply regulation, was previously designed using simulation studies. A dissolved oxygen monitoring system was installed into the facility, while nitrogen concentrations were analysed every day. Optimal dissolved oxygen profiles in the oxidation ditch were identified, while the air supply strategy was modified according to the variations in the influent. PMID- 14753568 TI - Alternatives for upgrading the Wilderness Wastewater Treatment Plant for biological nutrient removal. AB - The Wilderness Wastewater Treatment Plant (WWTP) located in Orange County, Virginia is a four concentric ring oxidation ditch activated sludge system with a rated capacity of 1,935 m3/day. The three outer rings are used for wastewater treatment and the inner ring is used as an aerobic digester. The flow capacity has been increased from 1,935 to 3,760 m3/d, however, the desired design capacity has since been increased to 3,870 m3/d, and there are plans to eventually expand to approximately 4,840 m3/d with improved nitrogen removal. The design goal for the planned upgrade is to discharge an effluent that contains less than 10 mg/l total nitrogen (TN) at all times, with an annual average of 8 mg/l or less. In this study, the pre-upgrade performance of the Wilderness Wastewater Treatment Plant was evaluated and several modifications were recommended for the incorporation of biological nutrient removal (BNR). PMID- 14753569 TI - Biofilm bacteria inactivation by citric acid and resuspension evaluations for drinking water production systems. AB - The study investigates the inactivation of biofilm bacteria colonized on the surface of polyvinyl chloride (PVC) pipes delivering either groundwater or treated wastewater. It does so using a citric acid (C6H8O7) solution. The results of the study showed that the optimal conditions of the biofilm bacteria inactivation were over 10,000 mg/L citric acid concentration and 60 minutes of contact time at least. Under these conditions, the removal efficiency could reach above 99.999% for heterotrophic plate count (HPC) bacteria and 99.95% for coliform bacteria. The study also showed that the biofilm bacteria were the major sources of planktonic bacteria resuspended into water purified by drinking water production systems (DWPS). PMID- 14753570 TI - Direct filtration with preozonation for small water treatment systems. AB - The possibility of applying preozonation followed by direct filtration without the use of a coagulant is investigated. Filtration experiments have been carried out using four different water sources within Istanbul. A 1 m deep bed of 0.8-1.2 mm silica sand was used as the filter medium. The filter was operated at a rate of 11.5 m/hr. Raw water turbidities ranged from 2.3 NTU to 10.0 NTU. Effluent turbidity and particle count values were monitored using an on-line turbidimeter and an on-line particle counter. With all the waters studied in this work, preozonation improved particle removal in direct filtration. With three of the mentioned waters, it was possible to achieve turbidity values below 1.0 NTU without a coagulant by applying ozone at a rate between 1.7-1.8 mg/L. It was not possible to reduce the turbidity of one of the waters to below 2.7 NTU even with very high ozone dosages. The need for pilot testing before deciding if this treatment method is applicable to a given water is underlined. PMID- 14753571 TI - Use of reflectors to enhance the synergistic effects of solar heating and solar wavelengths to disinfect drinking water sources. AB - Aluminum reflectors were added to solar units designed to inactivate faecal microorganisms (faecal coliform, E. coli, enterococci, FRNA coliphage, C. perfringens) in stream water and diluted sewage by the two mechanisms (solar heat, solar UV) known to inactivate microorganisms. During sunny conditions, solar units with and without reflectors inactivated E. coli to <1 CFU/100 ml to meet drinking water standards. Solar units with reflectors disinfected the water sooner by increasing the water temperature by 8-10 degrees C to 64-75 degrees C. However, FRNA coliphages were still detected in these samples, indicating that this treatment may not inactivate pathogenic human enteric viruses. During cloudy conditions, reflectors only increased the water temperature by 3-4 degrees C to a maximum of 43-49 degrees C and E. coli was not completely inactivated. Under sunny and cloudy conditions, the UV wavelengths of sunlight worked synergistically with increasing water temperatures and were able to disinfect microorganisms at temperatures (45-56 degrees C), which were not effective in inactivating microorganisms. Relative resistance to the solar disinfecting effects were C. perfringens > FRNA coliphages > enterococci > E. coli > faecal coliform. PMID- 14753572 TI - Biological denitrification of drinking water using various natural organic solid substrates. AB - Denitrification of drinking water was studied using various natural organic solid substrates (NOSS) such as poplar, hornbeam, pine shavings and wheat straw as a carbon source in a batch unit. The highest nitrate removal efficiency was observed with the wheat straw, so it was chosen as the carbon source for biodenitrification in an upflow laboratory reactor. In order to remove solid particles from the effluent water, a sand filter unit was placed after the denitrification reactor. The soluble DOC contents in the reactor affected the efficiency of nitrate elimination and nitrate concentration of the effluent water remained below acceptable values (50 mg/l NO3-). In order to remove colour, DOC and nitrate from the water, powdered activated carbon adsorption studies were performed in the batch unit. PMID- 14753573 TI - On the use of crushed shells of apricot stones as the upper layer in dual media filters. AB - The use of crushed shells of apricot stones instead of anthracite coal in dual media filters is investigated. Turbidity removal efficiencies were measured for several filtration rates in dual media filters composed of shells of apricot stones above silica sand and anthracite coal above silica sand. Backwash (fluidization) experiments were carried out using sieved fractions of crushed shells of apricot stones to establish curves of expansion versus backwash velocity at 25 degrees C. Such curves can be used in the design of filters employing this material. It is believed that the use of such a locally available and cheaper alternative filter medium would be of interest in many small water and wastewater treatment systems around the world. PMID- 14753574 TI - Kinetic study of reactions between ozone and benzothiazole in water. AB - Benzothiazoles are frequently present in wastewater from rubber related applications, and may be found in surface and underground water bodies causing significant environmental impact. Cost effective treatment processes to deal with such contaminants are needed in both small and large-scale applications. These compounds are poorly biodegradable and could be removed by ozone oxidation before discharge to recipient water courses. Unfortunately, there is limited experimental data reported in the literature on such processes involving benzothiazoles. This article presents experimental data on ozone treatment of benzothiazole (BT), with a view to process design. The effects of pH and radical scavengers on process rate and removal efficiency were assessed at bench scale. Experimental results show that BT could be effectively removed using ozonisation, particularly at pH above 4. The presence of free radical scavengers drastically reduced the BT removal rate even at very low concentrations. Both direct and indirect reactions between ozone and BT were adequately described by second order kinetic schemes, with rate constants estimated at 20 degrees C: kD = 2.3 mol l( 1) s(-1) and kI = 6 x 10(9) mol l(-1) s(-1), respectively. The free radical mechanism accounted for 83-96% of BT removal rate within the pH range 2-9, at 20 degrees C. PMID- 14753575 TI - Beta-endosulfan removal from water by ozone oxidation. AB - Beta-endosulfan solutions were ozonated in a lab scale semi-batch reactor in various experimental conditions. Ozonation kinetics of beta-endosulfan and effects of some parameters such as pH, temperature, partial pressure and ozone dosage on oxidation were investigated. Increasing ozone dosage and decreasing of temperature and pH increased the oxidation rate of beta-endosulfan. Maximum 97% of beta-endosulfan could be removed at both 16-mg/min ozone dosages and pH = 4 for 60 minutes of ozonation. The order of the reaction is determined as pseudo first order. Although the rate of reaction was lower than other pesticide oxidation rates cited in the literature, beta-endosulfan reaction to ozonation was achieved with almost 90% removal rate. The temperature dependent rate constant for beta-endosulfan oxidation was determined as kd = 0.947 exp(-2.16 x 10(-3)/T). PMID- 14753576 TI - Paediatric intravenous fluid regimens need review. PMID- 14753577 TI - Treatment of obsessive compulsive disorder: a review paper. PMID- 14753578 TI - A prospective analysis of inpatient consultations to a gastroenterology service. AB - The provision of a formal consultation service for inpatients between subspecialists is little studied. We prospectively surveyed the pattern of inpatient consultations from hospital-based generalists and surgeons to the gastroenterology (GI) service for inpatients in a large urban teaching hospital over a 5 month period. There are two GI consultants/attendings and five GI registrars/fellows on the service. A formal consultation is made by the requesting team to the GI service using the hospital computer network. All referrals over a 5 month period were prospectively analysed. 242 consecutive inpatient referrals were sent to the GI service over 5 months. Average age was 56 years, 48.8% males. 32 consultants/attendings from other disciplines sent referrals. Most patients were seen within one working day. Urgent referrals were seen without delay. The commonest reasons for referral were abdominal pain (15.8%), percutaneous endoscopic gastrostomy (PEG) tube insertion (13.6%), diarrhoea (12.8%), abnormal liver blood tests (10%), nausea and vomiting (8.2%), anaemia (6.2%), and melaena (4.9%). Iatrogenic diseases accounted for 6.2% of consultations. Ongoing patient care was assumed by the GI team in 9.5% of referrals. 15.3% required a second consultation visit before discharge. 22.7% of referrals were followed in the GI outpatients' clinic after discharge. 51.2% underwent an endoscopic procedure. 13.6% of referrals were for PEG tube insertion. A quarter of these were considered unsuitable for immediate PEG tube insertion. Subspecialty consultation provides an expert opinion, encourages discussion and learning, and improves patient care. In our experience, the provision of specialist advice and reassurance often speeded up a patient's work up and expedited discharge. However, evaluating referral patients and subsequently providing ongoing inpatient and outpatient care and provision of endoscopy for these referrals contributes significantly to the workload of the GI service. PMID- 14753579 TI - Percutaneous endoscopic gastrostomy: 5 years of clinical experience on 238 patients. AB - Since Percutaneous Endoscopic Gastrostomy (PEG) feeding was introduced, 20 years ago it has been increasingly utilised in medical practice. The aim of this study was to assess the current indications and complications associated with PEG feeding. This study was a retrospective review of hospital charts dealing with PEG placement over a period of five years. The indications for insertion were, central nervous disease 76% (n = 156), other benign disease 14% (n = 28) and malignancy 10% (n = 21). Cerebrovascular accidents (CVA) alone accounted for 47% (n = 97). Ninety seven (50%) patients had minor complications, which included 43 (22%) wound infections. There were 6 (3%) major complications, including peritonitis, perforation and aspiration pneumonia. There were four deaths (2%) related to PEG placement, of whom three developed aspiration pneumonia and one peritonitis. The overall 30 day mortality rate was 16%. There was a 75% increase in the use of PEG placement over the five year period. PEG placements were associated with a 53% morbidity and a 2% procedure related mortality. There was a 16% 30 day mortality following PEG placement suggesting that the selection criteria for PEG placement may need to be refined further. PMID- 14753580 TI - Achieving 95% uptake in an immunisation programme--not an impossibility. AB - This study sought to document uptake rates achieved for vaccinations administered in a School Immunisation Programme in a school year. In a population of 6436 children uptake rates of more than 95% were achieved for all the vaccines administered, except MMR in the 6th class cohort for which 94.7% was the uptake figure. The success of the programme was attributed to the fact that the programme was the responsibility of a dedicated team, which facilitated the active follow-up of all defaulters as identified by the comprehensive database made available by the schools. Avoidance of deferral for minor illness was also a key factor. PMID- 14753581 TI - Screening of lower back pain, low back pain clinic. The clinical experience. AB - Acute low back pain is a common complaint with high prevalence in society. Orthopaedic and specialist spinal services may be overwhelmed by large numbers of patients with low back pain who do not require investigation or surgical intervention. This phenomenon has led to the establishment of back pain screening clinics as a system of triage for those with acute low back pain. In March 2001, a back pain screening clinic was established in the Adelaide and Meath Hospitals incorporating the National Children's hospital. Six hundred and sixty-five patients were seen in the first year of this clinic. Of these, only ninety-six required referral to the orthopaedic spinal clinic. Just twenty-nine of these patients required surgical intervention. The introduction of this service resulted in a reduction in the waiting periods for the specialised spine clinic. We conclude that back pain screening clinics result in a clinical and economical improvement in the care of those suffering from acute low back pain. This is achieved by the systematic and efficient assessment, treatment and referral of patients afflicted with acute low back pain. PMID- 14753582 TI - Research misconduct--a review of current considerations. PMID- 14753583 TI - Paediatric near-drowning: mortality and outcome in a temperate climate. AB - The decision whether to continue to resuscitate the paediatric victim of near drowning is influenced by potential poor neurological outcome. A low core body temperature at presentation is frequently cited as a reason to continue resuscitation. We report the case of an 11 month old infant admitted to the intensive care unit following near-drowning and a prolonged resuscitation. The infant's core body temperature was 29 degrees C. Cardiac output was restored, but the child remains in a persistent vegetative state. We present the results of a ten year review of near-drowning in a tertiary referral institution, to evaluate the mortality and outcome in a temperate climate. Thirteen patients were identified in the review. The mortality was 23%. The incidence of a persistent vegetative state was 15%. Asystole, immersion time greater than 15 minutes, resuscitation time longer than 30 minutes, the administration of epinephrine, and a low core body temperature were associated with a poor outcome. PMID- 14753584 TI - Changes in body mass indices of patients with schizophrenia 3 years following the introduction of a weight management programme. PMID- 14753585 TI - Mastoiditis presenting as an acute abdomen with features of Lemierre's syndrome. PMID- 14753586 TI - Procedure clinic--the way forward? PMID- 14753587 TI - Adolescent alcohol misuse--searching for a solution. PMID- 14753588 TI - MRI-guided abdominal intervention. PMID- 14753589 TI - Demonstration of extensive mesenteric venous thrombosis and intestinal infarction with multidetector row CT: value of curved planar reformations. AB - We describe a case of extensive mesenteric venous thrombosis with small bowel infarction. Multidetector row computed tomography with curved planar reformation clearly demonstrated the full extents of the mesenteric venous thrombus and the infarcted bowel loops. The length of infarcted segment could be estimated from the curved planar reformatted image and correlated well with operative findings. To our knowledge, this is the first presentation and measurement of infarcted bowel loops on curved planar reformation. PMID- 14753590 TI - Value of virtual computed tomographic colonography for Crohn's colitis: comparison with endoscopy and barium enema. AB - BACKGROUND: Crohn's colitis, frequently accompanied by stenosis or narrowing, can be difficult to assess through conventional methods. We evaluated the usefulness of virtual computed tomographic colonography (CTC) for the detection of colonic lesions due to Crohn's disease. METHODS: Forty-two lesions in 33 patients with Crohn's disease were examined by CTC and barium enema (BE). Twenty-two patients also were examined by colonoscopy (CS). The visualization ability of CTC was compared with those of the other two methods. RESULTS: In the visualization of elevated lesions, there was no significant difference between CTC and BE (18 of 20, p = 0.487) or between CTC and CS (15 of 16, p = 0.99); however, ulcerative lesions were less often visualized by CTC. However, CTC enabled identification of serious lesions in the colon proximal to the stenosis in nine patients and was superior to BE and CS in terms of its ability to visualize the proximal site of the stenosis (p = 0.003). CONCLUSION: CTC is clinically useful for the evaluation of Crohn's colitis, especially those with stenotic lesions. PMID- 14753592 TI - Right colonic diverticulitis: MR appearance. AB - BACKGROUND: We evaluated the magnetic resonance (MR) features of right colonic diverticulitis. METHODS: This prospective study was based on five patients selected from a group of 156 patients admitted to the radiology department for further evaluation because of clinically suspected appendicitis. All five patients had ultrasound (US) and MR studies, and four patients also had computed tomography (CT). RESULTS: In all five patients, right-side diverticulitis was seen as an outpouching of the right colon with associated circumferential wall thickening of the colon and surrounding inflammatory changes. CONCLUSIONS: Our results suggest that MR imaging can be useful in the diagnosis of right colonic diverticulitis. An inflamed diverticulum with adjacent colonic wall thickening and surrounding inflamed fat are characteristic MR signs. MR imaging can be a valuable alternative to CT in young or pregnant patients who have suspected appendicitis and an equivocal US result. PMID- 14753591 TI - MR staging of primary colorectal carcinoma: comparison with surgical and histopathologic findings. AB - BACKGROUND: We retrospectively evaluated the accuracy of magnetic resonance (MR) imaging in staging colorectal cancer and assessing local tumor extent, nodal involvement, and distant abdominal and pelvic metastases. METHODS: Forty-eight patients with primary colorectal carcinoma were referred for presurgical abdominal and pelvic MR imaging. MR imaging included T1-weighted, fat-suppressed T2-weighted, and fat-suppressed gadolinium-enhanced spin gradient-echo imaging. The prospective interpretations of the MR examinations were reviewed. MR depiction of local tumor extent, nodal involvement, and distant metastases at 18 anatomic locations was noted and compared with subsequent surgical and histopathologic findings. RESULTS: Overall TNM MR staging agreed with surgical and pathologic staging in 41 (85%) of 48 patients, including 21 (78%) of 27 colon cancers and 20 (95%) of 21 rectal cancers. For depth of tumor penetration, which was evaluable in 44 patients, MR imaging agreed with pathologic results in 38 (86%) of 44 patients, including 22 (88%) of 25 colon cancers and 16 (84%) of 19 rectal cancers. In 42 (95%) of 44 patients, MR images correctly distinguished tumor confined to the bowel wall (T0, T1, and T2) from tumor with transmural tumor extension (T3 and T4). Regional nodal metastases were depicted in 15 of 22 patients (sensitivity, 68%; accuracy, 83%). Nodal metastases were better depicted for rectal cancer in eight of nine patients, compared with colon cancer in seven of 13 patients. Distant metastases were correctly depicted on MR imaging in 13 of 14 patients (sensitivity, 93%; accuracy, 98%). In the site-by-site analysis, MR imaging prospectively depicted 66 of 77 sites of surgically confirmed metastatic tumor in the abdomen and pelvis (sensitivity, 86%; specificity, 99%; accuracy, 98%). CONCLUSION: MR imaging using currently available techniques can effectively image local tumor extent and distant metastases in patients with colorectal carcinoma. Especially for colon cancer, incomplete depiction of nodal metastases in normal-size lymph nodes remains a limitation of cross-sectional imaging studies. PMID- 14753593 TI - Comparison of different injection forms in CT examination of the upper abdomen. AB - BACKGROUND: The optimal technique for administration of intravenous contrast medium in computed tomography (CT) remains controversial. Therefore, we analyzed the influence of variable-rate injection protocols. METHODS: A double-blind, parallel-group study was conducted in 60 patients studied with the same helical CT contrast-enhanced protocol. Patients were randomly distributed into three groups: monophasic (123 mL at 2.5 mL/s), biphasic (123 mL, 60 mL at 1.5 mL/s and then 63 mL at 2.5 mL/s), and sigmoid (0.6 mL/s ending at 2.5 mL/s). Contrast enhancement efficacy was evaluated by attenuation coefficient measurements. RESULTS: The monophasic injection protocol produced a statistically higher liver, inferior vena cava, and portal enhancement than did the low-high biphasic and sigmoid protocols. The biphasic protocol produced a statistically higher enhancement in the superior aorta. The enhancement obtained with the monophasic protocol was always higher than or equal to those obtained with the biphasic protocol in all measurement protocols except in the superior aorta and the aortic bifurcation. CONCLUSIONS: A monophasic injection produces better parenchymal and venous enhancement. When arterial enhancement is important, a low-high biphasic protocol can be used. A sigmoid protocol, with the parameters used in our series, is significantly less effective. PMID- 14753594 TI - CT fluoroscopy-guided catheterization of the celiac and mesenteric arteries for concurrent CTHA and CTAP. AB - We describe a technique for computed tomographic (CT) fluoroscopy-guided celiac artery or superior mesenteric artery (SMA) catheterization for use with CT hepatic arteriography or CT arterial portography, respectively. Patients underwent conventional hepatic angiography to define the anatomy and to place a catheter within the celiac artery or the SMA. Subsequently, the catheter was repositioned in the target vessels under CT fluoroscopy. Our success rate was 94%. PMID- 14753595 TI - Cystic changes in hepatic and peritoneal metastases from gastrointestinal stromal tumors treated with Gleevec. AB - BACKGROUND: Tyrosine kinase inhibitor (Gleevec or STI-571) must be considered the treatment of choice for metastatic gastrointestinal stromal tumors (GISTs). The purpose of this article is to address and illustrate a long-term follow-up of computed tomographic (CT) radiologic findings in patients with metastases from GIST after Gleevec treatment. METHODS: We performed a retrospective review of seven patients (four male, three female) with unresectable metastases from GIST who were treated with STI-571 in a 1-year period. Patients were followed every 2 4 months by contrast-enhanced CT for up to 12 months. The size and attenuation of hepatic and peritoneal metastases on CT were measured and correlated. RESULTS: Hepatic metastases from GISTs showed significant decreased attenuation from a mean of 60 HU to a mean of 32 HU (p < 0.01) in the first 2 months and continued decreasing attenuation to 23 HU at the 12-month follow-up. These metastases superficially resembled simple cysts. Most metastases became smaller, with more defined borders, after treatment. Histologic examination in a resected specimen revealed hepatic cyst with no residual tumor cells, regression of omental lesions, and extensive necrosis. CONCLUSIONS: CT findings of unresectable hepatic and peritoneal metastases from GIST displayed decreasing, near cystic attenuation and size as an effective regression in response to STI-571 treatment. PMID- 14753596 TI - Peritoneal serous papillary carcinoma: a reappraisal of CT imaging features and literature review. AB - We evaluated the imaging features of primary serous peritoneal carcinoma (PSPC) on computed tomography (CT) and reviewed the literature. Preoperative CT images of 11 women with PSPC were retrospectively reviewed. The clinical presentations and serum levels of CA-125 were recorded. Special attention was paid to the operative and histopathologic findings of the ovaries. Imaging features were correlated with those in the literature. An elevation of serum CA-125 was found in 91% of cases. The CT findings included ascites (82%), peritoneal nodules or masses (73%), and omental nodules or omental caking (64%). Absence of an overt ovarian mass was observed in 64% of cases. The clinical manifestations and imaging features in our patients were consistent with those in the literature. Eighty-five percent of the ovaries in our study were superficially involved by PSPC in histopathologic examination. In none of our cases could the ovarian size be clearly assessed in the CT images. In conclusion, the presence of diffuse peritoneal disease and the absence of an ovarian mass on CT and an elevation of serum CA-125 level is suggestive of PSPC. However, in our experience, the evaluation of ovarian morphology and size by CT alone may not be as reliable. PMID- 14753597 TI - Hemoperitoneum complicating snake bite: rare CT features. AB - Russels's viper bite victims with systemic poisoning may present with hemorrhagic manifestations, including spontaneous bleeding and incoagulable blood associated with disseminated intravascular coagulation and primary fibrinolysis, two of the most prominent manifestations of the systemic envenoming by this species. Various vascular complications of viperine snake bite have been reported in the literature. We report the computed tomographic findings in a case of snake bite resulting in hemoperitoneum, which, to the best of our knowledge, has not been reported in the literature. PMID- 14753598 TI - Mesenteric involvement in neurofibromatosis type 1: CT and MRI findings in two cases. AB - We report computerized tomographic and magnetic resonance imaging findings of neurofibromatosis type 1 with mesenteric involvement in two patients. The first patient was a 13-year-old female with a panmesenteric plexiform neurofibroma with segmental involvement of the bowel wall; she had a 3-year history of abdominal pain. The second patient was a 60-year-old female who presented with malignant transformation of multiple mesenteric neurofibromas 4 months after primary operation. PMID- 14753599 TI - Rare diffuse peritoneal malignant neoplasms: CT findings in two cases. AB - In the peritoneal cavity, diffuse serosal replacement by tumor is demonstrated usually by extensive carcinomatous involvement from gastric, colonic, or pancreatic tumors or less frequently by mesothelioma. Primary tumors other than mesothelioma are extremely rare in the peritoneum. The computed tomographic appearances of two cases of rare peritoneal tumors, epithelioid hemangioendothelioma and desmoplastic small round cell tumor, are described. PMID- 14753600 TI - Primary biliary cirrhosis complicated with sigmoid colonic varices: the usefulness of computed tomographic angiography. AB - Gastroesophageal varices are the major complication of portal hypertension. Ectopic varices develop in various organs such as the duodenum, colon, and gallbladder. However, varices other than at gastroesophageal or rectal sites is a rare entity. We report a 53-year-old patient with primary biliary cirrhosis complicated by sigmoid colonic varices. Computed tomographic angiography was useful to understand the entire status of the varices. PMID- 14753601 TI - Hepatic artery aneurysm, an unusual cause of obstructive jaundice: MR cholangiography findings. AB - Hepatic artery aneurysms are rare vascular lesions, often with a nonspecific clinical presentation, and difficult to diagnose before rupture. We report the case of a patient with obstructive jaundice caused by a compression on the right biliary duct by a right hepatic artery aneurysm in which the diagnosis was suggested by magnetic resonance cholangiography and confirmed by Doppler sonography and preoperative digital subtraction angiography. PMID- 14753602 TI - Choledochal cyst with malignancy: magnetic resonance imaging and magnetic resonance cholangiopancreatographic features in two cases. PMID- 14753603 TI - Primary biliary tract malignant melanoma: US, CT, and MR findings. AB - We report a case of primary biliary tract malignant melanoma occurring in a 47 year-old male. Ultrasonography and computed tomography showed multiple masses in the gallbladder and distal common bile duct that caused biliary tract dilatation. Magnetic resonance imaging showed that the polypoid masses in the gallbladder and common bile duct were of low signal intensity on T2-weighted images and of high signal intensity on unenhanced T1-weighted images. PMID- 14753604 TI - Endoscopic retrograde cholangiopancreatography in the evaluation of anomalous junction of the pancreaticobiliary duct and related disorders. AB - BACKGROUND: Anomalous junction of the pancreaticobiliary duct (AJPB) is a rare finding in endoscopic retrograde cholangiopancreatography (ERCP). We present our data on the incidence of AJPB and associated diseases. METHODS: A retrospective analysis of 2885 ERCPs performed over 15 years was done to study the incidence of AJPB and the diseases associated with this anomaly. RESULTS: Of the 2885 patients who underwent ERCP, AJPB was seen in 46 (1.6%). AJPB was taken as a common channel exceeding 15 mm in length with or without dilatation of the common channel. Anomalous junction was of type I in 50%, type II in 39.1%, and type III in 2.2% cases. Y-type anomalous junction was seen in 8.7% of cases. Choledochal cyst was found in 87% of patients. Other disorders associated with AJPB were gallstones, gallbladder polyps, gallbladder carcinoma, protein plugs. and pancreatic ductal calculi. Four patients with AJPB did not show any associated abnormality. CONCLUSION: AJPB is a rare finding, and the diagnosis is based on a common channel longer than 15 mm on ERCP. Choledochal cyst is the most common association with AJPB. We believe that the clinical spectrum of AJPB may unfold further with the widespread use of cholangiographic techniques. PMID- 14753605 TI - Focal fatty sparing of the pancreatic head in cystic fibrosis: CT findings. AB - The most common imaging appearance of the pancreas in cystic fibrosis is diffuse, complete fatty replacement. We present a case of complete fatty replacement of the pancreatic body and tail with total sparing of the pancreatic head. To our knowledge, this pattern of fatty sparing and its associated computed tomographic appearance have not been previously reported in cystic fibrosis. PMID- 14753606 TI - Heterotaxy syndrome with pancreatic malrotation: CT features. AB - We report the case of an asymptomatic elderly man with heterotaxy syndrome who presented with incomplete pancreatic rotation and an aplastic pancreatic uncinate process, polysplenia, nonrotation of the intestine, a midline liver, a midline gallbladder, a right-sided stomach, and interruption of the inferior vena cava with azygos continuation. To our knowledge, pancreatic malrotation with resultant oblique placement has not been described in patients with heterotaxy syndrome, and we speculate that the pancreatic rotation was interfered with by the left lobe of the midline liver in the embryo. PMID- 14753607 TI - Endoscopic ultrasound localization of a solitary insulinoma of pancreatic tail misdiagnosed as epilepsy: case report. AB - A 17-year-old female patient with features of epilepsy was treated with valproic acid. Two years later, hypoglycemia and hyperinsulinemia appeared. Transabdominal ultrasonography, spiral computed tomography, and indium-111 Octreoscan were performed without positive results. Endoscopic ultrasonography identified an oval tumor in the pancreatic tail with a color Doppler hypervascular pattern. Surgical enucleation decreased levels of insulin and C-peptide within 20 min, and the patient became free of symptoms and medications. PMID- 14753608 TI - Intrapancreatic accessory spleen: evaluation by CT arteriography. AB - BACKGROUND: Intrapancreatic accessory spleens are frequently confused with primary pancreatic tumors, and differentiation from neoplastic lesions is important to avoid an unnecessary laparotomy. We present three cases of intrapancreatic accessory spleen evaluated by computed tomographic arteriography (CTA) and discuss the characteristic findings. METHODS: CTA was performed, followed by digital subtraction angiography, with an injection of contrast material through a 4-F catheter placed in the celiac artery. Single-level dynamic CTA was also performed in two patients with a 30-s continuous scan in one breath hold. RESULTS: CTA clearly demonstrated early inhomogeneous enhancement of the lesion, similar to the splenic parenchyma. On single-level dynamic CTA, inhomogeneous enhancement of the lesion in the early phase was diminished in the late phase. Multiplanar reformatted images obtained in two cases showed the deep cleft between the lesion and the pancreas, which suggested that the lesion was originally extrapancreatic. CONCLUSIONS: These two findings on CTA, inhomogeneous enhancement of the lesion and the deep cleft between the lesion and the pancreas, may help to confirm the diagnosis of an intrapancreatic accessory spleen. PMID- 14753609 TI - Extensive abdominal splenosis: imaging features. AB - We present the case of a 22-year-old patient who had splenectomy in childhood after trauma and had a known chronic active infection with hepatitis C. Imaging procedures in different radiologic modalities diagnosed diffuse intraabdominal splenosis. Splenosis is a rare, severe complication of splenic trauma or surgery, but the pathogenesis is not clear. Imaging features play a key role in the diagnosis of ectopic splenic tissue, which must be differentiated from malignancies, especially from lymphoma. Splenosis itself may induce relapse of hematologic diseases, mainly autoimmune thrombocytopenia. PMID- 14753610 TI - Tumors of the urinary bladder: technique, current use, and perspectives of MR and CT cystography. AB - In comparison with other applications of virtual endoscopy, reports about virtual cystocopy are limited. In this update article, the various techniques of magnetic resonance (MR) and computed tomographic (CT) cystography are explained with their inherent advantages and disadvantages. The principal methods of postprocessing are presented, and future technical developments of MR and CT cystography are described. The present clinical applications and the possible role of MR and CT cystography in the future are discussed. PMID- 14753611 TI - MRI of fetal abdominal abnormalities. AB - Although ultrasonography is the method of choice for evaluating the fetus, magnetic resonance imaging (MRI) complements ultrasonography in the accurate diagnosis of fetal abnormalities. The advantages of MRI include excellent tissue contrast, a large field of view, and relative operator independence. To date, most studies on fetal MRI have focused on the fetal central nervous system and thoracic disorders. However, our experience suggests that MRI can be helpful even in evaluating fetal abdominal disorders. This pictorial essay illustrates the various MRI appearances of fetal abdominal abnormalities and discusses the indications and advantages of fetal MRI. PMID- 14753612 TI - Residual contrast medium in the cortex of the kidney around an infected renal cyst on CT: case report. AB - We present a case of a patient with an infected renal cyst. Delayed computed tomography showed residual contrast medium in the cortex of the kidney around it. Delayed computed tomography might be useful to identify an infected renal cyst. PMID- 14753613 TI - Glomerulocystic kidney disease: MRI findings. AB - Glomerulocystic kidney disease (GCKD) is a rare form of renal cystic disease characterized by cystic dilation of Bowman's capsule. The imaging findings of small renal cysts with a predominant cortical and subcapsular distribution allows for distinction from other, more common, polycystic kidney diseases. The appearance and distribution of the renal cysts by magnetic resonance imaging allow for a definitive diagnosis of GCKD. PMID- 14753614 TI - Endometrial ossification and infertility: the diagnostic value of different imaging techniques. AB - We present a case of longstanding secondary subfertility caused by endometrial ossification. Of all diagnostic techniques performed, magnetic resonance imaging and hysterosalpingography did not detect the abnormality. Transvaginal ultrasound and computed tomography clearly showed the endometrial pathology. After successful operative hysteroscopy with removal of the osseous tissue, the patient became pregnant spontaneously within 2 months. PMID- 14753615 TI - Endometriosis of the rectovaginal septum: endovaginal US and MRI findings in two cases. AB - We report two patients with endometriosis of the rectovaginal septum in whom endovaginal ultrasound (US) and magnetic resonance imaging (MRI) played crucial roles in the diagnosis, staging, and follow-up of the disease. The specific features of endometriosis of the rectovaginal septum on MRI and transvaginal US enable a noninvasive diagnosis, thereby reducing diagnostic delay and avoiding unnecessary invasive procedures. PMID- 14753616 TI - A VSA-based strategy for placing conservation buffers in agricultural watersheds. AB - Conservation buffers have the potential to reduce agricultural nonpoint source pollution and improve terrestrial wildlife habitat, landscape biodiversity, flood control, recreation, and aesthetics. Conservation buffers, streamside areas and riparian wetlands are being used or have been proposed to control agricultural nonpoint source pollution. This paper proposes an innovative strategy for placing conservation buffers based on the able source area (VSA) hydrology. VSAs are small, variable but predictable portion of a watershed that regularly contributes to runoff generation. The VSA-based strategy involves the following three steps: first, identifying VSAs in landscapes based on natural characteristics such as hydrology, land use/cover, topography and soils; second, targeting areas within VSAs for conservation buffers; third, refining the size and location of conservation buffers based on other factors such as weather, environmental objectives, available funding and other best management practices. Building conservation buffers in VSAs allows agricultural runoff to more uniformly enter buffers and stay there longer, which increases the buffer's capacity to remove sediments and nutrients. A field-scale example is presented to demonstrate the effectiveness and cost-effectiveness of the within-VSA conservation buffer scenario relative to a typical edge-of-field buffer scenario. The results enhance the understanding of hydrological processes and interactions between agricultural lands and conservation buffers in agricultural landscapes, and provide practical guidance for land resource managers and conservationists who use conservation buffers to improve water quality and amenity values of agricultural landscape. PMID- 14753617 TI - Trail impacts in Sagarmatha (Mt. Everest) National Park, Nepal: a logistic regression analysis. AB - A trail study was conducted in the Sagarmatha (Mt. Everest) National Park, Nepal, during 1997-1998. Based on that study, this paper examines the spatial variability of trail conditions and analyzes factors that influence trail conditions. Logistic regression (multinomial logit model) is applied to examine the influence of use and environmental factors on trail conditions. The assessment of trail conditions is based on a four-class rating system: (class I, very little damaged; class II, moderately damaged, class III, heavily damaged; and class IV, severely damaged). Wald statistics and a model classification table have been used for data interpretation. Results indicate that altitude, trail gradient, hazard potential, and vegetation type are positively associated with trail condition. Trails are more degraded at higher altitude, on steep gradients, in areas with natural hazard potential, and within shrub/grassland zones. Strong correlations between high levels of trail degradation and higher frequencies of visitors and lodges were found. A detailed analysis of environmental and use factors could provide valuable information to park managers in their decisions about trail design, layout and maintenance, and efficient and effective visitor management strategies. Comparable studies on high alpine environments are needed to predict precisely the effects of topographic and climatic extremes. More refined approaches and experimental methods are necessary to control the effects of environmental factors. PMID- 14753619 TI - Spatial modeling of harvest constraints on wood supply versus wildlife habitat objectives. AB - We studied the effects of spatial and temporal timber harvesting constraints on competing objectives of sustaining wildlife habitat supply and meeting timber harvest objectives in a boreal mixedwood forest. A hierarchical modeling approach was taken, where strategic and tactical level models were used to project blocking and scheduling of harvest blocks. Harvest block size and proximity, together with short- and long-term temporal constraints, were adjusted in a factorial manner to allow creation of response-surface models. A new measure of the habitat mosaic was defined to describe the emergent pattern of habitat across the landscape. These models, together with multiple linear regression, were used to provide insight on convergence or divergence between spatial objectives. For example, green-up delay (defined as time required before a harvest block adjacent to a previously logged block can be scheduled for harvest) had an adverse effect on the amount of annual harvest area that could be allocated and blocked spatially, and habitat supply responded in an opposite direction to that of wood supply, where caribou, moose wintering, and marten habitat supply increased when harvest blocks were further apart, maximum block size smaller, and both a green up delay and mesoscale stratification were applied. Although there was no "solution space" free of conflicts, the analysis suggests that application of the mesoscale stratification, together with a diversity of harvest block sizes and a between-harvest block proximity of 250 m, will perform relatively well with respect to wood supply objectives, and at the same time create a less fragmented landscape that better reflects natural forest patterns. PMID- 14753618 TI - Benchmark criteria: a tool for selecting appropriate models in the field of water management. AB - A milestone in the field of European water protection policy is the European Union's Water Framework Directive (WFD), which came into force in December 2000 and which integrates the management of European waters in many ways. In this study, we start by focusing on management issues connected to the implementation of the WFD and pose a question: "what type of models would be the most suitable for use in the context of the WFD?" With this question in mind, we aim to establish a set of operational and functional selection criteria for (computer) models whose application is intended to support decision-making related to a particular water management issue. These so-called "benchmark criteria" should help water managers and other model users in choosing appropriate models, e.g., for the WFD implementation purposes. We first describe models and their use in general and then propose an approach for setting the benchmark criteria for models, basing it on the concept of uncertainty management, while keeping firmly in mind the important role of citizens and citizen organizations in water management. The suggested benchmark criteria are in the form of 14 questions through which each model can be evaluated. Finally, the process for testing and refining the benchmark criteria is highlighted. PMID- 14753620 TI - Importance of sediment-water interactions in Coeur d'Alene Lake, Idaho, USA: management implications. AB - A field study at Coeur d'Alene Lake, Idaho, USA, was conducted between October 1998 and August 2001 to examine the potential importance of sediment-water interactions on contaminant transport and to provide the first direct measurements of the benthic flux of dissolved solutes of environmental concern in this lake. Because of potential ecological effects, dissolved zinc and orthophosphate were the solutes of primary interest. Results from deployments of an in situ flux chamber indicated that benthic fluxes of dissolved Zn and orthophosphate were comparable in magnitude to riverine inputs. Tracer analyses and benthic-community metrics provided evidence that solute benthic flux were diffusion-controlled at the flux-chamber deployment sites. That is, effects of biomixing (or bioturbation) and ground-water interactions did not strongly influence benthic flux. Remediation efforts in the river might not produce desired water-quality effects in the lake because imposed shifts in concentration gradients near the sediment-water interface would generate a benthic feedback response. Therefore, development of water-quality models to justify remediation strategies requires consideration of contaminant flux between the water column and underlying sediment in basins that have been affected by long-term (decadal) anthropogenic activities. PMID- 14753621 TI - Development and adoption of a simple nonpoint source pollution model for Port Phillip Bay, Australia. AB - New computing tools and approaches allow tailored development of software to meet the needs of environmental managers. The processes required for such tailoring fit well with adaptive management concepts where, as knowledge and system understanding develop among managers, the software can be developed or replaced to match. This paper reports on development and adoption of a simple nonpoint source pollution modeling tool, including technical aspects of data support for modeling and social aspects of software design. The software, named FILTER, used a unit load model to generate expected pollutant loads from subcatchments of Port Phillip Bay, Australia. Monitoring data were used for calibration to modify the delivery of generated pollutants to receiving waters. Spatial, tabular, and charting software components were used to provide alternative forms of output visualization. FILTER was developed using a process that resulted in manager stakeholders taking responsibility for setting of model parameter values and operation of the user interface, thereby encouraging uptake. The inclusive development process, tailoring of the software to manager needs and styles of usage, and matching of model complexity to data and knowledge, resulted in a successful application that has become the current agreed system representation among disparate stakeholder organizations. PMID- 14753622 TI - Influence of geoengineered climate on the terrestrial biosphere. AB - Various geoengineering schemes have been proposed to counteract anthropogenically induced climate change. In a previous study, it was suggested that a 1.8% reduction in solar radiation incident on the Earth's surface could noticeably reduce regional and seasonal climate change from increased atmospheric carbon dioxide (CO2). However, the response of the terrestrial biosphere to reduced solar radiation in a CO2-rich climate was not investigated. In this study, we hypothesized that a reduction in incident solar radiation in a Doubled CO2 atmosphere will diminish the net primary productivity (NPP) of terrestrial ecosystems, potentially accelerating the accumulation of CO2 in the atmosphere. We used a dynamic global ecosystem model, the Integrated Biosphere Simulator (IBIS), to investigate this hypothesis in an unperturbed climatology. While this simplified modeling framework effectively separated the influence of CO2 and sunlight on the terrestrial biosphere, it did not consider the complex feedbacks within the Earth's climate system. Our analysis indicated that compared to a Doubled CO2 scenario, reduction in incident solar radiation by 1.8% in a double CO2 world will have negligible impact on the NPP of terrestrial ecosystems. There were, however, spatial variations in the response of NPP-engineered solar radiation. While productivity decreased by less than 2% in the tropical and boreal forests as hypothesized, it increased by a similar percentage in the temperate deciduous forests and grasslands. This increase in productivity was attributed to an approximately 1% reduction in evapotranspiration in the Geoengineered scenario relative to the Doubled CO2 scenario. Our initial hypothesis was rejected because of unanticipated effects of engineered solar radiation on the hydrologic cycle. However, any geoengineering approaches that reduce incident solar radiation need to be thoroughly analyzed in view of the implications on ecosystem productivity and the hydrologic cycle. PMID- 14753623 TI - Depleting fish resources, declining fishing communities, and the state revitalization project in Korea. AB - This paper reviews recent declines in fish resources and fishing communities in Korea, evaluates major factors of declines, and examines a subsequent community based revitalization project. Factors leading to depleting fish resources include global regulations, technological advances in fishing, national economic development policies, land reclamation projects, and pollution. The negative economic and societal impact of depleting fish resources is addressed using demographic and income levels of the communities. In response, the state initiated revitalization projects and invested in local fishing cooperatives promoting community-based management. The result has been less than effective as it led to mismanagement of investment and created an attitude of dependence on the government for financial assistance. PMID- 14753624 TI - Hedonic analysis of effects of a nonnative invader (Myriophyllum heterophyllum) on New Hampshire (USA) lakefront properties. AB - Introduced species are a major threat to the planet's ecosystems and one of the major causes of species extinction. This study deals with some of the economic impacts of one of these "invaders," variable milfoil. Variable milfoil can clog water-bodies, cause boating and swimming hazards, and crowd out native species. This study analyzed the effects of variable milfoil on shoreline property values at selected New Hampshire lakes. Results indicate that property values on lakes experiencing milfoil infestation may be considerably lower than similar properties on uninfested lakes. Results are highly sensitive to specification (variable selection) of the hedonic equation. PMID- 14753625 TI - Spectrally driven classification of high spatial resolution, hyperspectral imagery: a tool for mapping in-stream habitat. AB - Streams represent an essential component of functional ecosystems and serve as sensitive indicators of disturbance. Accurate mapping and monitoring of these features is therefore critical, and this study explored the potential to characterize aquatic habitat with remotely sensed data. High spatial resolution, hyperspectral imagery of the Lamar River, Wyoming, USA, was used to examine the relationship between spectrally defined classes and field-mapped habitats. Advantages of this approach included enhanced depiction of fine-scale heterogeneity and improved portrayal of gradational zones between adjacent features. Certain habitat types delineated in the field were strongly associated with specific image classes, but most included areas of diverse spectral character; spatially buffering the field map polygons strengthened this association. Canonical discriminant analysis (CDA) indicated that the ratio of the variability among groups to that within a group was an order of magnitude greater for spectrally defined image classes (20.84) than for field-mapped habitat types (1.82), suggesting that unsupervised image classification might more effectively categorize the fluvial environment. CDA results also suggested that shortwave-infrared wavelengths were valuable for distinguishing various in stream habitats. Although hyperspectral stream classification seemed capable of identifying more features than previously recognized, the technique also suggested that the intrinsic complexity of the Lamar River would preclude its subdivision into a discrete number of classes. Establishing physically based linkages between observed spectral patterns and ecologically relevant channel characteristics will require additional research, but hyperspectral stream classification could provide novel insight into fluvial systems while emerging as a potentially powerful tool for resource management. PMID- 14753626 TI - The implications of genetic mutations in the sodium channel gene (SCN5A). AB - Mutations in sodium channel alpha-subunit gene (SCN5A) result in multiple arrhythmic syndromes, including long QT3 (LQT3), Brugada syndrome (BS), an inherited cardiac conduction defect, sudden unexpected nocturnal death syndrome (SUNDS) and sudden infant death syndrome (SIDS), constituting a spectrum of disease entities termed Na+ channelopathies. These diseases are allelic disorders, if not the same disease with variable penetrance and variable modifiers worldwide. Interestingly, death occurs during sleep in all of these disorders, suggesting a common mechanism. To date, mutational analyses have revealed about 103 distinct mutations in SCN5A, of which at least more than 30 mutations are associated with LQT3, whereas the rest of the mutations are affiliated with the remaining sodium channel disorders. The majority of these mutations are missense. However, other types such as deletions, insertions, frameshifts, nonsense and splice-donor errors have also been reported. PMID- 14753627 TI - Correlation between electrogram morphology and standard criteria to validate bidirectional cavotricuspid block in common atrial flutter ablation. AB - AIM: Assessment of a bidirectional conduction block within the cavotricuspid isthmus (CTI) is critical during radiofrequency (RF) atrial flutter (AF) ablation. We investigated the use of bipolar atrial electrogram (BAE) morphology as an additional criterion identifying CTI block and tested it against two recognized criteria: differential pacing and reversal of the right atrial depolarization sequence during coronary sinus (CS) pacing. METHODS AND RESULTS: An RF ablation procedure was performed during 600 ms CS pacing in 100 consecutive patients with a common AF. BAE recorded along the CTI were continuously monitored. CTI conduction block was achieved by RF ablation in all patients and a clear change in BAE polarity in the Electrogram recorded by the dipoles located on the CTI and immediately lateral to the intended line of block (RS to QR pattern) associated with a confirmed CTI conduction block was observed in all cases. BAE morphology changes predicted bidirectional CTI conduction blocks with a 100% positive and a 100% negative predictive value. At a mean follow-up of 33 +/- 11 months, there was a 5% AF recurrence rate. CONCLUSIONS: Our study suggests that morphological changes in BAE recorded at sites lateral and adjacent to the target line of block may be used as a unique and robust criterion to validate CTI conduction block during AF ablation procedure. PMID- 14753628 TI - Noninvasive, direct visualization of macro-reentrant circuits by using magnetocardiograms: initiation and persistence of atrial flutter. AB - AIMS: We analysed the cardiac magnetic fields on the body surface to visualize electrical currents noninvasively during reentrant arrhythmias. METHODS AND RESULTS: Seven patients with counterclockwise atrial flutter (AFL) were studied during 17 episodes of AFL using 64-channel magnetocardiograms (MCGs) and electrophysiological study. Eight of the episodes were paroxysmal AFL, in which MCGs were recorded from the time of spontaneous onset to the time of termination. We constructed iso-magnetic field maps of the tangential components and produced MCG animations. With respect to AFL initiation, an atrial premature complex induced AFL. Prior to the initiation of AFL, atrial fibrillation (AF) transiently occurred. The cardiac magnetic fields revealed a single peak during sinus rhythm or with premature complexes but a disorganized pattern during AF. When AF transformed to AFL, the magnetic fields changed from a disorganized pattern to a single peak at first and then evolved to a circular pattern. During persistent AFL, the magnetic source moved in a counterclockwise circuit. CONCLUSION: MCG animation can be used to visualize the sequence in which a premature complex transforms sinus rhythm to AFL via AF. Our findings indicate that MCGs can be used to identify noninvasively the mechanisms responsible for atrial tachyarrhythmias. PMID- 14753629 TI - Comparison of generic health survey SF-36 and arrhythmia related symptom severity check list in relation to post-therapy AF recurrence. AB - AIM: The effect of applied therapy on quality of life (QoL) in patients with atrial fibrillation (AF) was investigated in recent studies. However, no information on clinical relevance of QoL assessing instruments in relation to post-ablation recurrence of AF is currently available. The aim of this study was to evaluate the clinical relevance of SF-36 and Arrhythmia Related Symptom Severity Check List (SSCL) to post-procedure AF recurrences in patients with paroxysmal AF undergoing pulmonary vein isolation (PVI). METHODS AND RESULTS: Sixty consecutive patients with AF were enroled in the study. The QoL was measured using SF-36 scale and SSCL. The questionnaires were administered at baseline then 3, 6, 9 and 12 months after the procedure. In order to define statistical power in relation to AF recurrence the scores were dichotomized. Positive and negative predictive accuracy (PPA, NPA) and test efficiency (sum of PPA and NPA) were calculated. Twenty-one out of 60 patients experienced a total of 66 recurrences of AF during follow-up. The parameters of SF-36 provided maximum test efficiency of 1.36, whereas the test efficiency of SSCL was 1.79. CONCLUSION: We conclude that SSCL is more specific instrument for a measurement of PVI success or failure. PMID- 14753630 TI - How important is the assessment of quality of life after pulmonary vein isolation for paroxysmal atrial fibrillation? PMID- 14753631 TI - Pneumopericardium and pneumothorax contralateral to venous access site after permanent pacemaker implantation. AB - A 77-year-old female underwent implantation of a left-sided dual chamber permanent pacemaker for symptomatic bradycardia with active fixation leads. Eight hours after the procedure, the patient complained of shortness of breath and was found to have a 30% right pneumothorax on chest X-ray. Immediately, a chest tube was inserted, promptly relieving the symptoms. A CT scan of the chest revealed extrusion of the helix of the screw-in atrial lead, through the wall of the right atrial appendage. The helix was abutting a bulla in the right lung, the likely cause for pneumothorax and pneumopericardium. The atrial lead was explanted without incident. PMID- 14753632 TI - Isolation of atrial fibrillation to a pulmonary vein. PMID- 14753633 TI - Long QT syndrome patients may faint due to neurocardiogenic syncope. AB - AIMS: Syncope in long QT syndrome (LQTS) is expected to be due to Torsades de Pointes ventricular tachycardia (TdP). Often these patients faint in situations with emotional stress. The aim of the present study was to evaluate whether neurocardiogenic syncope occurs in LQTS. METHODS AND RESULTS: Ten untreated consecutive LQTS patients (age 11-72 years, median 37.5 years, five males and five females from five different families (one KvLQT1 mutation, two HERG mutations in three families and one without established genetic background)) were examined by a head-up tilt-table test (HUT). If syncope did not occur within 25 min, the patient received 0.25 mg nitroglycerine sublingually and the HUT was continued for 20 min. Nine out of 10 patients had a positive HUT. The syncope resulted from a combined vasodepressor and bradycardiac response. There were no cases of TdP. No syncope occurred in a 42-year-old asymptomatic male LQTS patient with a borderline prolonged QTc of 0.45 s and a HERG mutation. In 11 of 21 patients referred for syncope without LQTS a positive HUT was found (P < 0.10). CONCLUSION: Syncope in LQTS can be of neurocardiogenic origin and is not necessarily due to TdP. The reason for neurocardiogenic syncope in LQTS is unknown, but involvement of the autonomic nervous system outside the heart is possible. PMID- 14753634 TI - Unusual response to the ajmaline test in a patient with Brugada syndrome. AB - We present a Brugada syndrome patient who suffered an aborted sudden death. The ajmaline test (1 mg/kg body weight) induced accentuated alternans ST-segment elevation in V1-V2 without ventricular arrhythmias. It could represent silent ischaemia not detected before, failure of myocardial regions to repolarize in alternate beats due to transmural dispersion of conduction and refractoriness in the right ventricular outflow tract or a rate dependent sodium channel block by ajmaline. We need more studies to know whether this electrocardiographic sign is a risk factor for life-threatening ventricular arrhythmias in Brugada syndrome patients. PMID- 14753635 TI - Ablation of postinfarction ventricular tachycardia guided by isolated diastolic potentials. AB - Frequent recurrences of ventricular tachycardia (VT) despite implantable cardioverter-defibrillator (ICD) and antiarrhythmic drug therapy are a typical indication for catheter ablation. We performed endocardial mapping of an haemodynamically tolerated VT in a 67-year-old male patient. Isolated diastolic potentials (IDPs) of similar morphology were recorded during atrial paced rhythm at baseline and during monomorphic VT. The isolated potentials were required for initiation and maintenance of ventricular arrhythmia. These diastolic electrograms were considered to be part of the reentry circuit, as they remained constantly associated with VT during oscillations of cycle length and resetting. Validation of the ablation target was not performed by exact entrainment pacing in order to test the predictive value of the observed diagnostic phenomena. Radiofrequency (RF) energy applications were successful at the site where IDPs were recorded during atrial paced rhythm and VT. Ablation decreased the need for ICD therapies effectively in a patient with scar-related, slow VT. PMID- 14753636 TI - Implantable cardioverter defibrillator recipients: quality of life in recipients with and without ICD shock delivery: a prospective study. AB - AIMS: The experiences of patients who received shocks from their implantable cardioverter defibrillator (ICD) and how these events affect their overall adjustment are poorly understood. Our goal was to evaluate quality of life and psychological well-being, and the prevalence of, and changes in, depression and anxiety of patients who did or did not experience defibrillatory shocks in the first 12 months after ICD implantation. In total 167 patients were monitored after discharge. Four self-administered questionnaires were used. The first assessment took place before ICD implantation, the remaining three at 1, 6 and 12 months after discharge. RESULTS: We classified patients into three shock groups and one no shock group. A small group of ICD recipients (26%) received ICD shock delivery, usually during the last 6 months of the study. Borderline significant differences were found within the groups over time regarding physical role limitations (P < 0.051). Those who experienced shocks throughout the year (group 1) felt more limited in their daily activities due to physical or mental problems. All groups health (P < 0.001). Overall quality of life did not change significantly after 6 months. Anxiety and depression did not change significantly over time. In total 22-66% of patients reported clinically significant depressive symptoms throughout the first year, and 31-83% clinically significant symptoms of anxiety. ICD recipients who had experienced a shock were significantly more anxious one-year postdischarge than those who had not received a shock. CONCLUSIONS: Overall quality of life and psychological well-being did not change in ICD recipients, irrespective of whether they experienced defibrillatory shocks. The high prevalence of depression and anxiety can be interpreted as a response to the perceived physical and mental problems regarding daily activities. Our data indicate that patients who have experienced an ICD shock do not adapt well to living with an ICD, they are more anxious than ICD recipients who received no shocks. However, the anticipation of having another shock after experiencing one is less stressful than that of the first shock. We conclude that the lasting psychological distress will not dissipate spontaneously or naturally and that psychosocial interventions may be warranted. PMID- 14753638 TI - Potential device interaction of a dual chamber implantable cardioverter defibrillator in a patient with continuous spinal cord stimulation. AB - Spinal cord or thalamic deep brain stimulation with a pacemaker is becoming more important in the treatment of drug refractory pain due to peripheral vascular disease, angina pectoris and intractable tremor in patients with neurologic disorders such as Parkinson's disease. An additional indication for a cardiac pacemaker or implantable cardioverter defibrillator raises concerns about possible interactions between the implanted electrical devices. We report on a patient with existing spinal cord stimulation who survived sudden cardiac death and received a dual chamber cardioverter defibrillator capable of delivering tiered therapies in both the atrium and ventricle. PMID- 14753637 TI - Diazepam or midazolam for external DC cardioversion (the DORM Study). AB - AIMS: To compare the clinical efficacy and tolerability of two intravenous sedatives for physician-led DC cardioversion of atrial arrhythmias. METHODS AND RESULTS: One hundred and forty-one patients attending for elective DC cardioversion of atrial arrhythmias were randomized to intravenous midazolam or diazepam. Sedation was administered using titration protocols. Procedure times, operator satisfaction scores and adverse events were documented. The patients immediate, 24- and 48-h recall and awareness of after-effects were obtained from questionnaires. Seventy-one patients received midazolam (mean 12.5 mg) and 70 patients received diazepam (mean 28.1 mg). There were 16 minor adverse events with midazolam (20% hypotension, 3% oxygen desaturation) and nine with diazepam (7% hypotension, 6% required additional analgesia), P = 0.14. There were no major adverse events. Sedation time was 5.0 +/- 3.4 min for midazolam and 6.5 +/- 3.4 min for diazepam (P = 0.0016). Patients awoke 77 +/- 46 min post-sedation with midazolam and 39 +/- 24 min with diazepam (P < 0.0001). There was no recall of the procedure at 48 h and no difference in awareness of after-effects between the two groups at 24 or 48 h, P = ns. CONCLUSION: Physician-led cardioversion of atrial arrhythmias using intravenous sedation is effective and well tolerated. Sedation with diazepam was associated with fewer minor adverse events and a quicker recovery time than midazolam. PMID- 14753639 TI - Incremental programming of atrial anti-tachycardia pacing therapies in bradycardia-indicated patients: effects on therapy efficacy and atrial tachyarrhythmia burden. AB - AIMS: Efficacy of pace-termination of atrial arrhythmias (ATP) may depend on atrial cycle length and regularity. Whether device programming of ATP therapies can improve ATP efficacy and alter atrial tachyarrhythmia burden is unknown. METHODS AND RESULTS: ATP efficacy was evaluated in 61 patients (39 males; 66 +/- 10 years) with a standard indication for pacing, 95% with a history of AT/AF. Each patient was implanted with a novel DDDRP pacemaker capable of delivering ATP therapy. ATP efficacy and AT/AF frequency and burden were compared within each patient during a period of nominal ATP programming (NP) followed by a period of aggressive incremental programming (IP). Adjusted ATP-termination efficacy was higher during IP than during NP (54.8% vs 37.9%, P < 0.05). No differences in AT/AF frequency (3.3 +/- 5.9 vs 3.2 +/- 6.9 day(-1)) or burden (18 +/- 28% vs 18 +/- 29%) were observed comparing NP with IP. The majority of episodes during both the NP (81%) and IP (77%) periods terminated within 10 min. Episodes lasting 24 h or more accounted for only 0.4% of the episodes in both groups. but accounted for 38% of the average burden during NP and 51% during IP. CONCLUSIONS: Device programming of ATP therapies can influence the number of treated episodes and the efficacy of ATP therapies although arrhythmic frequency and burden may not change. Total atrial arrhythmia burden is disproportionately influenced by long (>24 h) episodes. PMID- 14753640 TI - Quality of life and exercise capacity in patients with prolonged PQ interval and dual chamber pacemakers: a randomized comparison of permanent ventricular stimulation vs intrinsic AV conduction. AB - AIMS: The aim of this study was to assess quality of life (QoL) and exercise capacity during permanent ventricular stimulation (PVS) compared with intrinsic atrioventricular conduction (IAVC) in patients with dual chamber pacemakers (PMs) and an intrinsic PQ interval >210 ms. Dual chamber PMs in patients with atrioventricular (AV) block are usually programmed to PVS in VDD or DDD mode, although IAVC is preserved, but prolonged. This results in PVS, although long periods of IAVC may occur. METHODS AND RESULTS: Fourteen consecutive patients (age 76 +/- 6 years; intermittent high degree AV block in six patients, binodal disease in eight patients) were enroled in a prospective, randomized, single blind, crossover study of IAVC vs PVC. To permit IAVC, programmed AV delays were prolonged. At the end of each phase, QoL scores were assessed using a questionnaire and echocardiography and cardiopulmonary stress tests were performed. During the study period with IAVC, 95 +/- 10% of the beats were conducted intrinsically. QoL scores (28.3 +/- 11 vs 29.3 +/- 13; P = 0.68), peak exercise capacity (5.4 +/- 2.4 vs 5.2 +/- 2.9 METs; P = 0.35) and peak oxygen uptake (19.8 +/- 4.5 vs 18.8 +/- 5.2 ml/kg/min; P = 0.16) were comparable during IAVC and PVS, respectively. Similar echocardiographic values were found for left ventricular (LV) ejection fraction (50 +/- 9% vs 51 +/- 10%; P = 0.67) and velocity time integral at the left ventricular outflow tract (24 +/- 5 vs 22 +/- 6 cm; P = 0.20), respectively. CONCLUSIONS: We conclude that in patients with dual chamber PMs and intermittent high degree AV block neither PVS nor IAVC is superior with respect to QoL or exercise capacity. Therefore, pulse generators may be programmed to IAVC to extend their longevity. PMID- 14753641 TI - Pericardial effusion and right-sided pneumothorax resulting from an atrial active fixation lead. AB - We report on a patient in whom an active-fixation pre-shaped atrial lead caused perforation of the right atrial wall, pericardium and pleura, resulting in pericardial effusion and right-sided pneumothorax. Chest X-ray did not demonstrate protrusion of the atrial lead outside the cardiac silhouette but computed tomography visualized the tip of the helix of the atrial screw-in electrode outside the contours of the right atrial appendage touching the right upper lobe of the lung. The lead was repositioned with resolution of pericardial effusion and pneumothorax. Due to their proximity to the right lung, high anterolaterally positioned atrial screw-in leads carry a small but definite risk for right-sided pneumothorax. PMID- 14753642 TI - The diagnostic value of ATP testing in patients with unexplained syncope. AB - A minority of patients with unexplained syncope has an increased susceptibility to adenosine triphosphate (ATP) injection. In these 'hypersensitive' patients, owing to its powerful cardiac and hypotensive effects, endogenous adenosine released under physiological and pathological conditions could trigger bradycardia and/or hypotension and cause syncope. This hypothesis still needs to be proven. However, there is some evidence that the ATP test identifies a group of patients with otherwise unexplained syncope with definite clinical features, absence of structural heart disease and benign prognosis. The mechanism of syncope is heterogeneous; indeed, in cases of electrocardiographic documentation of spontaneous syncope, either a long ventricular pause (mainly due to paroxysmal atrioventricular (AV) block) or no rhythm variations or even tachycardia were documented. ATP-positive patients have clinical features and mechanisms of syncope which are different from tilt-positive patients. Owing to its low positive predictive value, the ATP test is of little value in selecting treatment. A favourable outcome suggests a strategy of postponing treatment, in particular pacemaker therapy, until a definite diagnosis can be made by documenting a spontaneous syncopal relapse. PMID- 14753643 TI - Significance of syncope in patients with Alzheimer's disease treated with cholinesterase inhibitors. AB - We describe three cases of patients with Alzheimer's disease who presented with cardiac syncope soon after initiation of a cholinesterase inhibitor therapy (donepezil). Bradyarrhythmia was documented in two patients, considered probable in one, and was presumed related to the cholinergic therapy. Pacemaker implantation seemed justified rather than donepezil cessation. More over, it permitted an increase in donepezil dosage. PMID- 14753644 TI - Minimizing impacts of maintenance dredged material disposal in the coastal environment: a habitat approach. AB - At present, coastal disposal of maintenance dredged material constitutes one of the most important problems in coastal zone management and in some coastal areas represents the major anthropogenic disturbance to the benthos. In this review we first propose, based on the classic literature, that macrofaunal communities typical of environmentally stressed habitats are more resilient than those of more environmentally stable habitats, and we outline the macrofaunal successional changes following a disturbance. Second, from a review and analysis of the published and unpublished literature on macrofaunal recovery following maintenance dredged material deposition in the coastal environment, we compare the successional sequences and recovery rates in euhaline and polyhaline systems. The review reveals that invertebrate recovery following dredged material disposal in relatively unstressed marine environments generally takes between 1 and 4 years, while in more naturally stressed areas, recovery is generally achieved within 9 months, although deeper polyhaline habitats can take up to 2 years to recover. Differences in recovery times are attributed to the number of successional stages required to regain the original community composition and that species typical of naturally unstressed assemblages do not possess life history traits to allow rapid recolonization of disturbances. In the last section of this review, the management implications of these findings are discussed in terms of minimizing dredged material disposal impacts on fisheries resources. Since the natural disturbance regime appears to be very important in determining the response of a benthic community following dredged material disposal, it is recommended that when predicting the potential environmental impact of an operation, the nature of the physical environment in combination with the status (and role) of associated marine benthic communities should be considered. PMID- 14753645 TI - Tourist perceptions of degradation caused by coastal nature-based recreation. AB - Tourist perceptions of environmental degradation caused by nature-based tourism activities in a coastal environment were determined in the Central Coast Region of Western Australia. Structured surveys were administered to 702 visitors over two peak seasons. Visitors were required to indicate their perceptions on a Likert-type scale. Activities assessed were swimming, boating, fishing, diving and snorkelling, (wind)surfing, sandboarding, four-wheel driving, (bush)walking, camping, horseriding and sightseeing. Tourists had significantly variable demographic characteristics over two seasons and participated in different activities. However, perception of environmental degradation of individual activities did not vary significantly between seasons, except for fishing, four wheel driving and sandboarding. The age, origin and level of education of visitors had more effect on perceptions than gender or income group. Participation in an activity affected perceptions only for those who went fishing, sandboarding, four-wheel driving and sightseeing. Visitor perceptions were comparable to 'real' impacts documented in the recreation ecology literature. The results of this research indicate a need for improved visitor education and interpretation facilities. PMID- 14753646 TI - Managing a subsidized predator population: reducing common raven predation on desert tortoises. AB - Human communities often are an inadvertent source of food, water, and other resources to native species of wildlife. Because these resources are more stable and predictable than those in a natural environment, animals that subsist on them are able to increase in numbers and expand their range, much to the detriment of their competitors and species they prey upon. In the Mojave Desert, common ravens (Corvus corax) have benefited from human-provided resources to increase in population size precipitously in recent years. This trend has caused concern because ravens prey on juvenile desert tortoises (Gopherus agassizi), a federally threatened species. In this paper, I discuss management strategies to reduce raven predation on desert tortoises. The recommendations fall into three categories: (1) managing raven populations by reducing access to anthropogenic resources; (2) removing offending ravens or other birds in specially targeted tortoise management zones; and (3) continuing research on raven ecology, raven behavior, and methods of reducing raven predation on tortoises. I also recommend approaching the problem within an adaptive management framework: management efforts should first be employed as scientific experiments--with replicates and controls--to yield an unbiased assessment of their effectiveness. Furthermore, these strategies should be implemented in concert with actions that reduce other causes of desert tortoise mortality to aid the long-term recovery of their populations. Overall, the approaches outlined in this paper are widely applicable to the management of subsidized predators, particularly where they present a threat to a declining species of prey. PMID- 14753647 TI - Decision support for road system analysis and modification on the Tahoe National Forest. AB - The United States Forest Service is required to analyze road systems on each of the national forests for potential environmental impacts. We have developed a novel and inexpensive way to do this using the Ecosystem Management Decision Support program (EMDS). We used EMDS to integrate a user-developed fuzzy logic knowledge base with a grid-based geographic information system to evaluate the degree of truth for assertions about a road's environmental impact. Using spatial data for natural and human processes in the Tahoe National Forest (TNF, California, USA), we evaluated the assertion "the road has a high potential for impacting the environment." We found a high level of agreement between the products of this evaluation and ground observations of a TNF transportation engineer, as well as occurrences of road failures. We used the modeled potential environmental impact to negatively weight roads for a least-cost path network analysis to 1573 points of interest in the forest. The network analysis showed that out of 8233 km of road analyzed in the forest, 3483 km (42%) must be kept in a modified road network to ensure access to these points. We found that the modified network had improved patch characteristics, such as significantly fewer "cherry stem" roads intruding into patches, an improved area-weighted mean shape index, and larger mean patch sizes, as compared to the original network. This analysis system could be used by any public agency to analyze infrastructure for environmental or other risk and included in other mandated analyses such as risks to watersheds. PMID- 14753648 TI - Measuring plant diversity in the tall threetip sagebrush steppe: influence of previous grazing management practices. AB - In July 2000, a 490-ha wildfire burned a portion of a long-term grazing study that had been established in 1924 at the US Sheep Experiment Station north of Dubois, Idaho, USA. Earlier vegetation measurements in this tall threetip sagebrush (Artemisia tripartita spp. tripartita) bunchgrass plant community documented significant changes in vegetation due to grazing and the timing of grazing by sheep. A study was initiated in May 2001 using 12 multiscale modified Whittaker plots to determine the consequences of previous grazing practices on postfire vegetation composition. Because there was only one wildfire and it did not burn all of the original plots, the treatments are not replicated in time or space. We reduce the potential effects of psuedoreplication by confining our discussion to the sample area only. There were a total of 84 species in the sampled areas with 69 in the spring-grazed area and 70 each in the fall- and ungrazed areas. Vegetation within plots was equally rich and even with similar numbers of abundant species. The spring-grazed plots, however, had half as much plant cover as the fall- and ungrazed plots and the spring-grazed plots had the largest proportion of plant cover composed of introduced (27%) and annual (34%) plants. The fall-grazed plots had the highest proportion of native perennial grasses (43%) and the lowest proportion of native annual forbs (1%). The ungrazed plots had the lowest proportion of introduced plants (4%) and the highest proportion of native perennial forbs (66%). The vegetation of spring-grazed plots is in a degraded condition for the environment and further degradation may continue, with or without continued grazing or some other disturbance. If ecosystem condition was based solely on plant diversity and only a count of species numbers was used to determine plant diversity, this research would have falsely concluded that grazing and timing of grazing did not impact the condition of the ecosystem. PMID- 14753649 TI - Comparison of four Sesbania species to remediate Pb/Zn and Cu mine tailings. AB - A 6-month greenhouse pot trial was performed, aimed at screening appropriate Sesbania species for remediation of Pb/Zn and Cu mine tailings. Performances of young seedlings of four Sesbania species (S. cannabina, S. grandiflora, S. rostrata, and S. sesban) were compared with and without inoculation of rhizobia. Seedlings were planted in two types of tailings amended with garden soil or garden soil mixed with river sediment. The results indicated that inoculated plants generally produced a higher biomass than samples without inoculation. Pb/Zn mine tailings containing rather high concentrations of total and water soluble Cu, Pb, and Zn were toxic to plant growth compared with Cu mine tailings, according to the growth performance of the four species. Sesbania sesban and S. rostrata showed superior growth performance, compared to the other two species. Thus, they can serve as pioneer species to modify the barren environment, by providing organic matter and essential nutrients such as nitrogen, upon decomposition, in a relatively short period of time. This is especially true for S. rostrata, which is an annual plant that forms both stem and root nodules. However, a longer-term field trial should be conducted to investigate if superior species can beneficially modify the habitat for the growth of subsequent plant communities. PMID- 14753650 TI - Field-based evaluation tool for riparian buffer zones in agricultural catchments. AB - Riparian buffer zones can improve water quality and enhance habitat, but a comprehensive yet rapid method that can assist the resource manager in assessing the effectiveness of buffers is not available. The aim of this paper is to describe and illustrate the use of a newly developed field-based evaluation tool for riparian buffer zones in agricultural catchments. The Buffer Zone Inventory and Evaluation Form (BZIEF) incorporates criteria-based scoring systems developed from literature review, subsequent peer-review, and then a pilot field study. Use of the BZIEF is demonstrated by comparing buffer zones in three catchments established for water quality and habitat improvement under the Water Fringe Option agrienvironment scheme in England in order to assess whether the buffers were likely to provide environmental enhancement. Results among the three catchments were generally similar; buffer zones scored highly for their abundant vegetation cover, lack of erosion, stream habitat quality, and sufficient width. Furthermore, previous grassland or arable land use did not substantially affect buffer zone ratings. However, the BZIEF indicated that inappropriate soil characteristics in one catchment were likely to constrain buffer zone effectiveness for improving water quality. In another catchment, poor riparian vegetation diversity and structure may yield ineffective habitat enhancement, according to the BZIEF. It was concluded that the BZIEF might be a useful tool for buffer zone comparison and monitoring, even though more work is needed to test and validate the method. For example, the BZIEF could be used to target appropriate locations for buffer zones and is flexible, so could be adapted for different policies, objectives and regions. PMID- 14753651 TI - Use of radar remote sensing (RADARSAT) to map winter wetland habitat for shorebirds in an agricultural landscape. AB - Many of today's agricultural landscapes once held vast amounts of wetland habitat for waterbirds and other wildlife. Successful restoration of these landscapes relies on access to accurate maps of the wetlands that remain. We used C-band (5.6-cm-wavelength), HH-polarized radar remote sensing (RADARSAT) at a 38 degree incidence angle (8-m resolution) to map the distribution of winter shorebird (Charadriiformes) habitat on agricultural lands in the Willamette Valley of western Oregon. We acquired imagery on three dates (10 December 1999, 27 January 2000, and 15 March 2000) and simultaneously collected ground reference data to classify radar signatures and evaluate map accuracy of four habitat classes: (1) wet with < or = 50% vegetation (considered optimal shorebird habitat), (2) wet with > 50% vegetation, (3) dry with < or = 50% vegetation, and (4) dry with > 50% vegetation. Overall accuracy varied from 45 to 60% among the three images, but the accuracy of focal class 1 was greater, ranging from 72 to 80%. Class 4 coverage was stable and dominated maps (40% of mapped study area) for all three dates, while class 3 coverage decreased slightly throughout the study period. Among wet classes, class 1 was most abundant (30% coverage) in December and January, decreasing in March by 15%. Conversely, class 2 increased dramatically from January to March, likely due to transition from class 1 as vegetation grew. This approach was successful in detecting optimal habitat for shorebirds on agricultural lands. For modest classification schemes, radar remote sensing is a valuable option for wetland mapping in areas where cloud cover is persistent. PMID- 14753652 TI - Utilization of photographs for determining impact indicators for trail management. AB - "Indicators" are a key component of resource management, but selecting them can be difficult and controversial. This study explores different methods for selecting impact indicators for management of trail conditions by comparing photo survey methods (PSM) with written survey methods (WSM). A total of 661 subjects participated in interviews conducted within Mudeung-Mountain Provincial Park in Kwang-ju, Korea. PSM explained variation in acceptability better than WSM. Six major photo components (in order of importance: area of large rocks on trail, area of wood steps, area of bare roots, area of bare soil, area of small stones, and area of stone steps) explained 84.7% of the variation in acceptability of trail management conditions. This study suggests that PSM can be a useful tool for selecting impact indicators. PMID- 14753653 TI - Decision support system for managing oil spill events. AB - The Mediterranean environment is exposed to various hazards, including oil spills, forest fires, and floods, making the development of a decision support system (DSS) for emergency management an objective of utmost importance. The present work presents a complete DSS for managing marine pollution events caused by oil spills. The system provides all the necessary tools for early detection of oil-spills from satellite images, monitoring of their evolution, estimation of the accident consequences and provision of support to responsible Public Authorities during clean-up operations. The heart of the system is an image processing-geographic information system and other assistant individual software tools that perform oil spill evolution simulation and all other necessary numerical calculations as well as cartographic and reporting tasks related to a specific management of the oil spill event. The cartographic information is derived from the extant general maps representing detailed information concerning several regional environmental and land-cover characteristics as well as financial activities of the application area. Early notification of the authorities with up-to-date accurate information on the position and evolution of the oil spill, combined with the detailed coastal maps, is of paramount importance for emergency assessment and effective clean-up operations that would prevent environmental hazard. An application was developed for the Region of Crete, an area particularly vulnerable to oil spills due to its location, ecological characteristics, and local economic activities. PMID- 14753654 TI - The International Alliance of ALS/MND Associations. AB - The Alliance is an active participant in the support of people living with ALS/MND throughout the world. In partnership with the many others who are part of the fight against ALS/MND, the aim of the Alliance is to share the expertise, knowledge and skills of ALS/MND Associations. For more complete information on The Alliance please visit our web site at alliance@alsmndalliance.org. If you would like assistance in establishing an ALS/MND patient care organization where one currently does not exist, contact the Alliance Secretariat at alliance@alsmndalliance.org and request a new organization pack. If you would like to contribute in some other way, contact the Secretariat and they will refer you to the most appropriate member of the Board of Directors. Zoe Tebbutt- Secretariat, PO Box 246 Northampton NN1 2PR, UK, Tel: +44 1604 611 821, Fax: +44 1604 611 852, E-mail: alliance@alsmndalliance.org, Internet: www.alsmndalliance.org. The Alliance Annual Meeting is the most important meeting of patient care organizations in the world. In 2003 it will be held in Milan, Italy, on November 14 and 15, followed by the International Symposium, November 17-19. Observers are welcome and encouraged. PMID- 14753655 TI - Psychosocial factors and cognition in amyotrophic lateral sclerosis. PMID- 14753656 TI - The genetics of motor neuron diseases. AB - Motor neuron diseases may be divided into three categories: those with lower motor neuron involvement--spinal muscular atrophy (SMA) and spinobulbar muscular atrophy (SBMA or Kennedy's disease); those with upper motor neuron involvement- primary lateral sclerosis (PLS) and the spastic paraplegias; and those with combined upper and lower motor neuron involvement--amyotrophic lateral sclerosis (ALS). Other familial motor neuron disorders include hereditary neuronopathies, GM2 gangliosidosis, and possibly the ALS/PD syndrome of Guam. The contribution of genetics to the etiopathogenesis of motor neuron considerably, accounting for a high percentage of spinal muscular atrophies, but only a small fraction of cases of ALS. The mode of inheritance also varies, with examples of autosomal dominant (AD), autosomal recessive (AR), or X-linked kindreds. (Tables 1 and 2). PMID- 14753657 TI - Relevance of oxidative injury in the pathogenesis of motor neuron diseases. AB - Oxidative stress, which results from a complex interplay of pro- and anti-oxidant forces, is generally considered to be the major effector of accumulation of oxidatively modified protein accumulation in cells, although reduced degradation due to impairment of proteolytic activity could also contribute. The discovery that a familial lateral sclerosis (ALS) results from mutations in the gene encoding Cu/Zn superoxide dismutase a anti-oxidant enzyme, stimulated considerable evaluation of reactive oxygen species (ROS) generation and oxidative protein damage in both familial and sporadic forms of the disease. Mutations in SOD1 do not cause disease by compromising dismutating activity, but through some toxic gain of function. Although exacerbation of other copper-catalyzed enzymatic activities has been demonstrated in vitro, there is little evidence substantiating that this property is responsible for toxicity in vivo. Studies of ROS generation and oxidative damage in vivo have produced mixed results, but collectively are consistent with oxidative stress playing a secondary role in pathogenesis of the disease. Studies of post-mortem tissue from sporadic ALS patients has yielded more consistent evidence of accumulation of oxidative damage to proteins, lipids, and DNA, but the time course of accumulation cannot be determined and the initiating causes of the disease have not been identified. The interplay between motor neurons and glial cells is important in the clinical progression of both familial and sporadic motor neuron diseases and release of reactive oxygen and nitrogen species or cytokines from microglia could contribute to the demise of motor neurons. This review describes the general mechanisms of radical-mediated cellular damage followed by the evidence for and against the contribution of oxidative injury to the pathogenesis of motor neuron diseases. PMID- 14753658 TI - Neuroimaging in amyotrophic lateral sclerosis. AB - Several neuroimaging modalities have been used with varying success to aid the clinical process of establishing the diagnosis of amyotrophic lateral sclerosis (ALS). By demonstrating evidence of occult upper motor neuron degeneration in vivo, a speedier and more definitive diagnosis in suspected cases could lead to earlier treatment and earlier enrollment in clinical trials. Findings compatible with ALS on routine MRI are not consistently found and are non-specific. Thus, routine anatomic imaging is useful in ruling out diseases that mimic ALS, but not in classification of new cases. Functional imaging techniques, such as PET and fMRI, have provided fascinating insights into the cortical functional reorganization that accompanies muscular weakness. PET and SPECT have revealed involvement of regions of the brain beyond the motor cortex, something not well appreciated by pathological examination. Of great need is a surrogate marker of therapeutic efficacy to make drug evaluation more efficient; neuroimaging, and magnetic resonance spectroscopy in particular, holds great promise in this regard in addition to helping us better understand the of neurodegeneration. PMID- 14753659 TI - Paradigms for the identification of new genes in motor neuron degeneration. AB - It is estimated that between 10-20% of amyotrophic lateral sclerosis (ALS) is familial and these cases encompass recessive and dominant modes of inheritance. So far, mutations in three genes, superoxide dismutase 1 (SOD1), the p150 subunit of dynactin (DCTN1), and alsin have been shown to be directly causal for motor neuron degeneration in humans. However, clearly the disorder is genetically heterogeneous and other causal genes remain to be found that explain the vast majority of familial ALS cases. Human genetics can be problematical in that it is difficult to detect linkage in disorders in which multiple loci give similar phenotypes and where families are often small. In addition, the vertical collection of generations is often not possible with late onset disorders. An excellent genetic model of humans is provided by the mouse. We can use mouse models of neurodegeneration to find new genes in the human population. These models are not exact replicas of the human condition, but are the mouse equivalent and are incredibly valuable resources for highlighting genes and biochemical pathways disrupted in ALS and other diseases. In addition mouse models give us access to both control and affected tissues, at all stages of development and disease, thus greatly facilitating our understanding of pathogenesis. They also provide us with model systems for testing new therapies. Here we describe the approach taken to the characterization of new models of motor neuron disease and illustrate this with examples, including a recently characterized mouse model, Legs at odd angles (Loa). PMID- 14753660 TI - A survey of clinicians' practice in the symptomatic treatment of ALS. AB - Symptomatic management is the mainstay of ALS patient care, but there are few controlled trials of drugs and interventions for common symptoms. METHODS: We queried ALS clinic neurologists to determine drugs and interventions of choice and neurologists' perceived efficacy for 14 symptoms. RESULTS: The results are tabulations of the physicians' four most frequent choices for each symptom, the physicians' perception of efficacy, the average doses and average daily costs. A wide range of drugs and interventions were nominated for management of ALS symptoms. Consensus on treatment was rare for individual symptoms, and efficacy for any symptom was judged moderate at best. A few drugs were recommended for multiple symptoms. Comparisons of perceived efficacy compared to drug costs are informative. DISCUSSION: The results of the survey emphasize the challenges of symptom management in ALS. These data aid clinical management and guide rational choices for randomized controlled trials. PMID- 14753661 TI - Hirayama's disease: a case report. PMID- 14753662 TI - Amyotrophic lateral sclerosis (ALS) and mirror movements in a patient with polymicrogyria. AB - We report on a case of polymicrogyria with benign childhood epilepsy and amyotrophic lateral sclerosis (ALS). While performing dexterity tasks with either hand, strong unsustained mirror movements of the unintended hand were observed. The patient was seen over a period of three years and, as often seen in ALS, there was a moderate progress of the motor neuron disease affecting the upper and lower motor neuron in an asymmetrical manner. In addition, more rapidly progressive bulbar symptoms could be observed by the clinical and neurophysiological examination. MRI revealed polymicrogyria of the right frontal lobe with irregular bumpy inner and outer surface and abnormally thick and smooth cortex, dysplastic insular cortex and asymmetrically widened Sylvial fissure. No signs of atrophy, especially of the motorcortex and corticospinal tract were observed. The corpus callosum was completely developed and of normal size. We hypothesize an enrolment of the dysplastic right frontal lobe pathophysiology of the observed mirror movements. PMID- 14753663 TI - What is noninvasive ventilation? PMID- 14753664 TI - System for the generation of standard gas mixtures of volatile and semi-volatile organic compounds for calibrations of solid-phase microextraction and other sampling devices. AB - Standard gases are used for quality control and quality assurance, development of analysis methods and novel air sampling devices. The use of solid-phase microextraction (SPME) and other novel technologies for research in the area of air sampling and analysis requires systems/devices for reliable standard gas generation and sampling. In this paper we describe a new gas standard generating system for volatile organic compounds (VOCs) and semi-VOCs that was designed, built, and tested to facilitate fundamental and applications research with SPME. The system provided for the generation of a wide range of VOC/semi-VOC concentrations and mixing various standard gases, estimation of detection limits, testing the effects of sampling time, air temperature and relative humidity, testing the effects of air velocity and ozone on sampling/extractions. The system can be also used for calibrations of analytical instrumentation, quality control and quality assurance checks, and cross-validations of SPME with/and other sampling techniques. PMID- 14753665 TI - Diffusion-based calibration for solid-phase microextraction of benzene, toluene, ethylbenzene, p-xylene and chlorobenzenes from aqueous samples. AB - Short-term solid-phase microextraction (SPME) was performed to test a recently proposed semi-empirical model for the prediction of concentrations of analyte in water samples from the fibre-extracted mass without further calibration. The mass uptake rates obtained for benzene, toluene, ethylbenzene and p-xylene (BTEX) differ considerably from the before published, showing that interfibre comparability is a serious issue. The relative prediction errors are between -55% for benzene and +82% for p-dichlorobenzene under optimal conditions, i.e. they are by an order of magnitude higher than originally published. A sensitivity analysis shows the dominant influence of the estimated thickness of the diffusional boundary layer around the fibre on the concentration predicted. Empirical modification of the model equation for this parameter yields satisfactory results under the conditions tested for both BTEX and the selected chlorobenzenes. PMID- 14753666 TI - Polydimethylsiloxane rod extraction, a novel technique for the determination of organic micropollutants in water samples by thermal desorption-capillary gas chromatography-mass spectrometry. AB - A novel, simple and inexpensive approach to absorptive extraction of organic compounds from environmental samples is presented. It consists of a polydimethylsiloxane rod used as an extraction media, enriched with analytes during shaking, then thermally desorbed and analyzed by GC-MS. Its performance was illustrated and evaluated for the enrichment of sub- to ng/l of selected chlorinated compounds (chlorobenzenes and polychlorinated biphenyls) in water samples. The new approach was compared to the stir bar sorptive extraction performance. A natural ground water sample from Bitterfeld, Germany, was also extracted using both methods, showing good agreement. The proposed approach presented good linearity, high sensitivity, good blank levels and recoveries comparable to stir bars, together with advantages such as simplicity, lower cost and higher feasibility. PMID- 14753667 TI - Development of miniaturized sample preparation with fibrous extraction media. AB - Introducing fine polymeric filaments as the extraction medium, a miniaturized sample preparation technique for micro-column liquid chromatography (micro-LC) has been developed along with the investigation of a reproducible preparation scheme of the extraction capillary. The polymeric filaments were packed longitudinally into either a fused-silica capillary or a polyether ether ketone (PEEK) capillary of appropriate dimensions, and the extraction capillary was installed to the injection valve in micro-LC system. The number of packed filaments should be precisely counted before the packing process to make sure the reproducible preparation of the extraction capillary. With conventional stationary phase materials for open-tubular gas chromatography, polymeric coating to the surface of the filaments was also studied in order to further enhance the extraction performance and selectivity. Coated with the polymeric material suitable for the extraction of particular analyte, a dramatic improvement on the extraction power was obtained. The results suggest that the future possibility of novel tailored fibrous extraction medium with an appropriate coating on it, especially for the analysis of complex sample matrices. PMID- 14753668 TI - Permeation liquid membrane for trace metal speciation in natural waters. Transport of liposoluble Cu(II) complexes. AB - In this paper, the transport of Cu(II) in the presence of lipophilic Cu(II) organic complexes through permeation liquid membranes (PLMs) have been investigated. In natural waters, small organic compounds, which form liposoluble neutral complexes with Cu(II), are potentially toxic and bioavailable. Hence, to understand the role of liposoluble Cu(II) complexes in natural waters, four organic ligands: phthalic acid, bipyridyl, pyrocatechol and hydroxyquinoline, which form uncharged or lipophilic Cu(II) complexes, were tested. The results showed that the transport of lipophilic Cu(II) complexes through PLM depends on the lipophilicity of the complex. Applications of PLMs in natural waters are presented. PMID- 14753669 TI - Comparison of different trapping methods for pressurised hot water extraction. AB - Four trapping methods for pressurised hot water extraction were compared in terms of recovery and selectivity. Also, robustness, repeatability and solvent consumption of the trapping systems were investigated. The trapping methods were collection into solvent following liquid-liquid extraction, solid-phase trapping into Tenax TA (SPE), flat sheet microporous membrane liquid-liquid extraction and hollow fibre microporous membrane liquid-liquid extraction. Polycyclic aromatic hydrocarbons were extracted with these systems from four soil and sediment matrices and the extracts were analysed by GC-MS and size-exclusion chromatography. Clear differences were observed in the selectivity and extraction efficiencies of the trapping systems. PMID- 14753670 TI - Steam distillation-solid-phase microextraction for the detection of Ephedra sinica in herbal preparations. AB - A new method involving concurrent solid-phase microextraction combined with continuous hydrodistillation of essential oil was developed. This new methodology allowed for the detection by GC-MS of very small amounts of a diagnostic peak for the authentication of Ephedra sinica, in a short period of time and using only small sample sizes. This diagnostic peak was identified as 4-vinylanisole, and elucidated from the chromatographic profile allowed for the identification of a sample as E. sinica among other species investigated in this study. To the best of our knowledge this is the first report on using continuous solid-phase microextraction coupled to hydrodistillation for the investigation of essential oil components, and the first report of 4-vinylanisole as a marker compound for E. sinica. A total of 46 collections representing 21 species of Ephedra were studied. PMID- 14753671 TI - Air sampling of aromatic hydrocarbons in the presence of ozone by solid-phase microextraction. AB - Effects of ozone on air sampling of standard gas mixtures of aromatic hydrocarbons were tested using solid-phase microextraction (SPME). Standard concentrations of ozone ranging from 10 ppb (v/v) to 6400 ppm (v/v) were generated using an in-house built ozone generator based on corona discharge. Effects of temperature, discharge voltage, and oxygen flow on the ozone generation were tested. The working dc voltage had the greatest effect on generated ozone concentration and was proportional to the ozone concentration. Generation temperature and oxygen flow rate were inversely proportional to ozone concentrations. Produced ozone was mixed with standard benzene, toluene, ethylbenzene, and xylenes (BTEX) gas at less than 100 ppb (v/v). Air samples were collected with poly(dimethylsiloxane) (PDMS) 100 microm SPME fibers and analyzed by gas chromatography (GC)-flame ionization detection (FID) and GC-MS. Significant reductions of BTEX concentrations were observed. In addition, some products of BTEX-ozone-oxygen reactions were identified. SPME worked well as a rapid sampler for BTEX and BTEX-ozone-oxygen reaction products. No significant deterioration of the PDMS coating and no significant reduction of absorption capacity were observed after repeated exposure to ozone. PMID- 14753672 TI - Sampling and analysis of volatile organic compounds in bovine breath by solid phase microextraction and gas chromatography-mass spectrometry. AB - A relatively noninvasive method consisting of a face mask sampling device, solid phase microextraction (SPME) fibers, and a gas chromatography-mass spectrometry (GC-MS) for the identification of volatile organic compounds (VOCs) in bovine breath was developed. Breath of three morbid steers with respiratory tract infections and three healthy steers were sampled seven times in 19 days for 15 min at each sampling. The breath VOCs adsorbed on the divinylbenzene (DVB) Carboxen-polydimethyl siloxane (PDMS) 50/30 microm SPME fibers were transported to a laboratory GC-MS system for separation and identification with an in-house spectral library of standard chemicals. A total of 21 VOCs were detected, many of them for the first time in cattle breath. Statistical analyses using Chi-square test on the frequency of detection of each VOC in each group was performed. The presence of acetaldehyde (P < or = 0.05) and decanal (P < or = 0.10) were associated more with clinically morbid steers while methyl acetate, heptane, octanal, 2,3-butadione, hexanoic acid, and phenol were associated with healthy steers at P < or = 0.10. The results suggest that noninvasive heath screening using breath analyses could become a useful diagnostic tool for animals and humans. PMID- 14753673 TI - Improvements in the methylmercury extraction from human hair by headspace solid phase microextraction followed by gas-chromatography cold-vapour atomic fluorescence spectrometry. AB - Improvements in the methylmercury extraction from human hair by solid-phase microextraction followed by gas chromatography coupled to cold-vapour atomic fluorescence spectrometry (GC-CVAFS) have been carried out. They consisted in the optimisation of the digestion step prior to the aqueous-phase ethylation and in the GC-CVAFS interface set-up. The main digestion parameters such as acid type, concentration, temperature and time have been optimised for hair sample analysis, thereby avoiding methylmercury degradation. Moreover, the stability of the digested samples was evaluated to improve the sample throughput. PMID- 14753674 TI - Full automation of derivatization--solid-phase microextraction-gas chromatography mass spectrometry with a dual-arm system for the determination of organometallic compounds in aqueous samples. AB - The determination of organometallic compounds in aqueous samples by in-vial derivatization and headspace solid-phase microextraction (SPME)-gas chromatography (GC)-mass spectrometry (MS) has been fully automated using a Twin PAL dual-arm robotic system. Linearity, accuracy, sensitivity for a series of n methyl, n-ethyl, and n-phenyl metal substituted chloride compounds of tin, lead, and mercury were investigated. The automated method was compared to similar manual methods and improved precision, speed and throughput was achieved. By originally programming the Twin PAL dual-arm system with the supplier's software (Cycle Composer, Version 1.5.0) the arms on the robot were only able to work in sequence. However, in order to have a flexible system and exploit time efficiently the robotic arms must work simultaneously. This was accomplished by programming the robot with the new software package called Cruise Control 4-2 for Twin PALs. Compared to Cycle Composer, Cruise Control 4-2 enhanced the speed and throughput of the automated system further. In addition, with a built-in crash prevention protocol and an improved user interface a more user-friendly system was obtained. PMID- 14753675 TI - Monitoring of drugs and metabolites in whole blood by restricted-access solid phase microextraction coupled to liquid chromatography-mass spectrometry. AB - Robust biocompatible solid-phase microextraction (SPME) devices were prepared using various alkyldiol-silica (ADS) restricted-access materials (RAM) as the SPME coating. The ADS-SPME approach was able to simultaneously fractionate the protein component from a biological sample, while directly extracting diazepam and the major metabolites N-desmethyldiazepam, oxazepam and temazepam, and overcame the present disadvantages of direct sampling in biological matrices by SPME. The devices were interfaced with an LC-MS system and an isocratic mobile phase was used to desorb, separate, and quantify the analytes. The calculated diazepam, nordiazepam, temazepam, and oxazepam detection limits were 20, 20, 30, and 35 ng/ml in heparinized blood, respectively. The method was confirmed to be linear over the range of 50-1000 ng/ml with an average linear coefficient (R2) value of 0.996. The injection repeatability and intra-assay precision of the method were evaluated over ten injections at concentrations of 50, 200, and 500 ng/ml, resulting in a R.S.D. of ca. 10%. The robustness of the ADS-SPME device was evaluated for future use in in vivo studies, providing many direct extractions and subsequent determination of benzodiazepines in blood. For the extraction of the peptides angiotensin I, II, and III from blood, a novel restricted access material with cation exchange properties was evaluated. The ion exchange diol silica improved the extraction efficiency of peptides relative to the conventional ADS material with reversed phase extraction centers. PMID- 14753676 TI - Comparison of different extraction methods for the analysis of volatile secondary metabolites of Lippia alba (Mill.) N.E. Brown, grown in Colombia, and evaluation of its in vitro antioxidant activity. AB - Hydrodistillation (HD), simultaneous distillation solvent extraction (SDE), microwave-assisted hydrodistillation (MWHD), and supercritical fluid (CO2) extraction (SFE) were employed to isolate volatile secondary metabolites from fresh leaves and stems of Colombian Lippia alba (Mill.) N.E. Brown. Kovats indices, mass spectra or standard compounds were used to identify around 40 components in the various volatile fractions. Carvone (40-57%) was the most abundant component, followed by limonene (24-37%), bicyclosesquiphellandrene (5 22%), piperitenone (1-2%), piperitone (ca. 1.0%), and beta-bourbonene (0.6-1.5%), in the HD, SDE, MWHD, and SFE volatile fractions. Static headspace (S-HS), simultaneous purge and trap in solvent (CH2Cl2) (P&T), and headspace solid-phase microextraction (HS-SPME) were used to sample volatiles from fresh L. alba stems and leaves. The main components isolated from the headspace of the fresh plant material were limonene (27-77%), carvone (14-30%), piperitone (0.3-0.5%), piperitenone (ca. 0.4%), and beta-bourbonene (0.5-6.5%). The in vitro antioxidant activity of L. alba essential oil, obtained by hydrodistillation was evaluated by determination of hexanal, the main carbonyl compound released by linoleic acid subjected to peroxidation (1 mm Fe2+, 37 degrees C, 12 h), and by quantification of this acid as its methyl ester. Under the same conditions, L. alba HD-essential oil and Vitamin E exhibited similar antioxidant effects. PMID- 14753677 TI - Analysis of volatile secondary metabolites from Colombian Xylopia aromatica (Lamarck) by different extraction and headspace methods and gas chromatography. AB - Hydrodistillation (HD), simultaneous distillation-solvent extraction (SDE), microwave-assisted hydrodistillation (MWHD), and supercritical fluid (CO2) extraction (SFE), were employed to isolate volatile secondary metabolites from Colombian Xylopia aromatica (Lamarck) fruits. Static headspace (S-HS), simultaneous purge and trap (P&T) in solvent (CH2Cl2), and headspace (HS) solid phase microextraction (SPME) were utilised to obtain volatile fractions from fruits of X. aromatica trees, which grow wild in Central and South America, and are abundant in Colombia. Kovats indices, mass spectra or standard compounds, were used to identify more than 50 individual components in the various volatile fractions. beta-Phellandrene was the main component found in the HD and MWHD essential oils, SDE and SFE extracts (61, 65, 57, and ca. 40%, respectively), followed by beta-myrcene (9.1, 9.3, 8.2 and 5.1%), and alpha-pinene (8.1, 7.3, 8.1 and 5.9%). The main components present in the volatile fractions of the X. aromatica fruits, isolated by S-HS, P&T and HS-SPME were beta-phellandrene (53.8, 35.7 and 39%), beta-myrcene (13.3, 12.3 and 10.1%), p-mentha-1(7),8-diene (7.1, 10.6 and 10.4%), alpha-phellandrene (2.2, 5.0 and 6.4%), and p-cymene (2.2,4.7 and 4.4%), respectively. PMID- 14753678 TI - Studies on the aroma of cupuassu liquor by headspace solid-phase microextraction and gas chromatography. AB - Headspace solid-phase microextraction (HS-SPME) coupled to gas chromatography with ion trap mass spectrometric detection and with atomic emission detection (GC AED) was employed to identify possible odor-impact volatile organic compounds in cupuassu (Theobroma grandiflorum Spreng) liquor, as well as to quantify alkylpyrazines present in these samples. SPME fibers coated with 100 microm polydimethylsiloxane (PDMS), 65 microm PDMS-divinylbenzene (DVB) and 75 microm Carboxen (CAR)-PDMS were tested, the later being chosen for the optimized extraction procedure. The principal compounds found in the sample headspace were 3-methylbutanal, dimethylsulfide, dimethyldisulfide, beta-linalool and several alkylpyrazines (notably tetramethylpyrazine). The procedure for quantitation of the alkylpyrazines, using GC-AED for their separation and detection, allowed the detection of microg g(-1) levels of the analytes in the samples, with acceptable precision (R.S.D. less than 10%). PMID- 14753679 TI - Physico-chemical measurements of CL-20 for environmental applications. Comparison with RDX and HMX. AB - CL-20 is a polycyclic energetic nitramine, which may soon replace the monocyclic nitramines RDX and HMX, because of its superior explosive performance. Therefore, to predict its environmental fate, analytical and physico-chemical data must be made available. An HPLC technique was thus developed to measure CL-20 in soil samples based on the US Environmental Protection Agency method 8330. We found that the soil water content and aging (21 days) had no effect on the recoveries (>92%) of CL-20, provided that the extracts were kept acidic (pH 3). The aqueous solubility of CL-20 was poor (3.6 mg l(-1) at 25 degrees C) and increased with temperature to reach 18.5 mg l(-1) at 60 degrees C. The octanol-water partition coefficient of CL-20 (log KOW = 1.92) was higher than that of RDX (log KOW = 0.90) and HMX (log KOW = 0.16), indicating its higher affinity to organic matter. Finally, CL-20 was found to decompose in non-acidified water upon contact with glass containers to give NO2- (2 equiv.), N2O (2 equiv.), and HCOO- (2 equiv.). The experimental findings suggest that CL-20 should be less persistent in the environment than RDX and HMX. PMID- 14753680 TI - Fast solid-phase extraction-gas chromatography-mass spectrometry procedure for oil fingerprinting. Application to the Prestige oil spill. AB - A rapid and simple fractionation procedure using solid-phase extraction (SPE) cartridges was developed for an accurate determination of aliphatic and polycyclic aromatic hydrocarbons in petroleum residues and further application in chemical fingerprinting of oil spills by gas chromatography-mass spectrometry (GC MS). Among the adsorbents evaluated, SiO2/C3-CN exhibited the best selectivity, providing, by elution with n-hexane (4 ml) and n-hexane-CH2Cl2 (1:1) (5 ml), two well-resolved aliphatic and aromatic hydrocarbon fractions, with recoveries of 97 +/- 7.2 and 99.7 +/- 13.9%, respectively. The SPE fractionation procedure was compared with the conventional silica-alumina adsorption chromatography showing similar results but practical advantages in terms of reproducibility, analysis time, solvent reduction and cost. Moreover, is particularly suitable for routine analysis with a high sample throughput. The developed methodology was tested in the characterization of fuel-oil samples collected along the Spanish north-west coast, after the Prestige oil spill accident. PMID- 14753681 TI - Mobility-based selective on-line preconcentration of proteins in capillary electrophoresis by controlling electroosmotic flow. AB - A simple method to perform selective on-line preconcentration of protein samples in capillary electrophoresis (CE) is described. The selectivity, based on protein electrophoretic mobility, was achieved by controlling electroosmotic flow (EOF). A short section of dialysis hollow fiber, serving as a porous joint, was connected between two lengths of fused silica capillary. High voltage was applied separately to each capillary, and the EOF in the system was controlled independently of the local electric field intensity by controlling the total voltage drop. An equation relating the EOF with the total voltage drop was derived and evaluated experimentally. On-line preconcentration of both positively charged and negatively charged model proteins was demonstrated without using discontinuous background electrolytes, and protein analytes were concentrated by approximately 60-200-fold under various conditions. For positively charged proteins, positive voltages of the same magnitude were applied at the free ends of the connected capillaries while the porous joint was grounded. This provided a zero EOF in the system and a non-zero local electric field in each capillary to drive the positively charged analytes to the porous joint. CE separation was then initiated by switching the polarity of the high voltage over the second capillary. For negatively charged proteins, the procedure was the same except negative voltages were applied at the free ends of the capillaries. Mobility based selective on-line preconcentration was also demonstrated with two negatively charged proteins, i.e. beta-lactoglobulin B and myoglobin. In this case, negative voltages of different values were applied at the free ends of the capillaries with different values, which provided a non-zero EOF in the system. The direction of EOF was the same as that of the electrophoretic migration velocities of the protein analytes in the first capillary and opposite in the second capillary. By controlling the EOF, beta-lactoglobulin B, which has a higher mobility, could be concentrated over 150-fold with a 15 min injection while myoglobin, which has a lower mobility, was eliminated from the system. PMID- 14753682 TI - AT-column, a novel concentrating technique for large-volume injections in gas chromatography. AB - Nowadays, large-volume injection is widely used for the GC determination of trace analytes, specifically to improve detectability. The most popular injectors for large-volume injections are the programmable temperature vaporisation (PTV) injector and the cold on-column (COC) injector, where each device has its own advantages and limitations. The novel AT-column concentrating technique combines features of two other injection techniques, loop-type large-volume and vapour overflow. AT-column injection is based on solvent evaporation in an empty liner with solvent vapour discharge via the split line. Little or no optimisation is required. The only relevant parameter is the injection temperature which can easily be calculated using the equation of Antoine. As an application, AT-column injection is combined with GC-MS for the trace-level determination of labile analytes and with GC-flame ionisation detection for the analysis of high molecular weight polymer additives. In summary, AT-column is an injection technique that combines the inertness of the COC, and the flexibility and robustness of the PTV large-volume technique. PMID- 14753683 TI - Speciation of butyltin compounds in marine sediments by preconcentration on C60 and gas chromatography-mass spectrometry. AB - A new method for the speciation of butyltin compounds by solid phase extraction and direct injection using gas chromatography-mass spectrometry (GC/MS) is described. The compounds were complexed with sodium diethyldithiocarbamate and retained on a C60 sorbent column. The neutral chelates of butyltin compounds were eluted with ethyl acetate containing NaBPr4 as derivatising reagent. The main analytical figures of merit of the proposed method for 10 ml of sample are: linear range 0.2-35 ng/g expressed as Sn; limits of detection, 0.07, 0.09 and 0.10 ng/g as Sn for monobutyltin, dibutyltin and tributyltin, respectively. No interferences from metal ions such as Zn2+, Fe3+, Sb3t, Pb2+, Ni2+ and Mn2+ were observed in the determination of organotin compounds. The validation of method was performed out by the analysis of a standard reference sediment (CRM 462). The method was also applied to the determination of butyltin compounds in marine sediment samples. PMID- 14753684 TI - Simultaneous solid-phase extraction of acidic, neutral and basic pharmaceuticals from aqueous samples at ambient (neutral) pH and their determination by gas chromatography-mass spectrometry. AB - Seven polymeric solid-phase extraction (SPE) sorbents were evaluated with regard to their ability to extract acidic, neutral and basic pharmaceuticals and estrogens simultaneously from water at neutral pH. Highest recoveries (70-100%) for the majority of the analytes were obtained with styrene-methacrylate and styrene-N-vinylpyrrolidone co-polymers. The latter one (Oasis HLB) was chosen for further refinement of an extraction method for the quantitative determination of acidic and neutral drugs in surface water samples at detection limits below 1 ng/l. A sequential elution protocol was applied for clean-up and separation of the extracted analytes into fractions suitable for further compound specific processing. The neutral analytes as well as the acidic compounds after derivatisation were quantified by GC-MS. Caffeine, ibuprofen, its metabolites and diclofenac were detected in river water samples in the 1-100 ng/l range. PMID- 14753685 TI - Drop formation in aqueous two-phase systems. AB - Extraction using aqueous two-phase systems (ATPSs) is a versatile technique for the downstream processing of various proteins/enzymes. The study of drop formation deals with the fundamental understanding of the behavior of liquid drops under the influence of various external body as well as surface forces. These studies provide a basis for designing of the extractions in column contactors in which liquid drops play a major role. Most of the drop formation studies reported so far is restricted to aqueous-organic systems. ATPSs, differ from aqueous-organic systems in their physical properties. In view of this, an attempt was made to develop a model for drop formation in ATPSs adopting the information available on aqueous-organic systems. In order to validate the model, experiments were performed by using polyethylene glycol (PEG)/salt systems of different phase compositions at various flow rates. At low flow rates the single stage model and at high flow rates the two stage model are able to predict the drop volume during its formation from tip of capillary. The experimental results were found to agree reasonably well with those predicted by the model. PMID- 14753686 TI - Twin column chromatography for industrial-scale decontamination processes. AB - A frontal chromatographic unit was devised consisting of a column-detector-column array. The unit is either equipped with identical columns (identical twins) or with columns of varying length (fraternal twins). Due to the finite nature of the columns, a prerun is formed at the column walls following the same regularities as the main stream. These regularities are used for the identification of the process termination below the detection limits of the monitor. For the implementation, a clear preference is given to the employment of fraternal twins, as the feed assay can be integrated into the separation process. PMID- 14753687 TI - Comparisons of the interactions between two analytes and two structurally similar chiral stationary phases using high-performance liquid chromatography, suspended state high-resolution magic angle spinning nuclear magnetic resonance and solid state nuclear magnetic resonance spectroscopy. AB - A study of the retention behaviour of the enantiomers (R)- and (S)-1,1' binaphthyl-2,2'-diol as well as (+) and t-)-O,O'-dibenzoyl-tartaric acid was performed using the two chiral stationary phases (CSPs) Kromasil-DMB and Kromasil TBB. Detailed information about the interactions between the analytes and the two CSPs was obtained from suspended-state HR/MAS transferred NOESY NMR experiments as well as suspended-state HR/MAS 1H NMR titration experiments. Good correlation between the suspended-state HR/MAS NMR experiments and the corresponding HPLC experiments was obtained. This shows that suspended-state HR/MAS NMR as well as solid-state CP/MAS NMR spectroscopy can be used to investigate interactions between stationary phases and analytes under conditions that are similar to those used in HPLC. PMID- 14753688 TI - Calculation of the molar absorptivity of polyphenols by using liquid chromatography with diode array detection: the case of carnosic acid. AB - Antioxidant activity of vegetable extracts is related to the nature and the amount of active components, mainly polyphenols; therefore, a correct quantification of these molecules should be required to define their concentration in such kind of vegetable extracts. A fast and accurate method to calculate molar absorption coefficients (epsilon), by using HPLC, has been tested on standard polyphenols and caffeine, and should be widely adapted for standardless quantitative analysis. Molar absorptivity (epsilon) of carnosic acid (CA) was determined from 200 to 300 nm, by the proposed method and those values were compared to tert-butyl-hydroxytoluene (BHT) ones for further comparative quantification. PMID- 14753689 TI - Comparative analysis of enzymatically digested kappa-carrageenans, using liquid chromatography on ion-exchange and porous graphitic carbon columns coupled to an evaporative light scattering detector. AB - Enzymatically digested kappa (A-G4S)-carrageenans, apart from their biological activities in plants, could be used as 'model' molecules to elucidate potential problems in nuclear magnetic resonance spectroscopy of carrageenans. Thus, oligosaccharides obtained from kappa-carrageenan by enzymatic digestion using kappa-carrageenase have been separated on silica and polymeric based ion-exchange and porous graphitic carbon (PGC) columns, coupled to an evaporative light scattering detector. Oligomers were separated on ion-exchange columns using a gradient of ammonium acetate as a developing ion, while analysis on PGC column presented an additional adjacent peak next to each main one, using a gradient of ammonium acetate in water/acetonitrile as a mobile phase. The phenomenon can be attributed to different retention mechanisms that govern the PGC surface. Furthermore, it has been demonstrated that acetonitrile can regulate the selectivity between the peaks raising hopes for preparative chromatography. PMID- 14753690 TI - Ion chromatographic separation of hydrogen ion and other common mono- and divalent cations. AB - We introduced an approach to the ion chromatographic determination of common mono and divalent cations including hydrogen ion and demonstrated the ability of a C30 column dynamically coated first with dodecylsulfate and then with 18-crown-6 ether to separate the cations by ion-exchange mechanism. Using an ethylenediamine solution containing a small concentration of 18-crown-6 ether and lithium dodecylsulfate at pH 6.2 as eluent, the cations were eluted in the order Li < Na+ < NH4+ < H+ < K+ < Mg2+ < Ca2+ with symmetrical peaks. The conductivity vs. concentration plots were linear about three orders of magnitude, from millimolar to micromolar; and the detection limits were all < 0.6 microM. Rainwater was analyzed directly using this ion chromatographic system with satisfactory results. PMID- 14753691 TI - Prediction of relative response factors of electron-capture detection for some polychlorinated biphenyls using chemometrics. AB - The relative response factor (RRF) of an electron-capture detection (ECD) system is predicted for a set of 118 polychlorinated biphenyls (PCBs). Due to the wide range of relative retention times of PCB congeners, the RRFs of these compounds were calculated based on two different internal standards. Therefore, the compounds were divided into two molecular subsets. As a first step, multiple linear regression (MLR) was employed to find informative descriptors that can predict the RRFs of these compounds. Two descriptors of molecular ion ionization potential (MIIP) and ionization potential of the molecule (IP) that are related to affinity of the compounds for the electrons show the highest mean effects in subsets 1 and 2. respectively. The descriptors appearing in the MLR models were considered as inputs for developing the back-propagation artificial neural networks (BP-ANN). Two networks with the architectures of 5-5-1 and 7-6-1 were generated for the prediction of RRFs of molecules of subsets 1 and 2, respectively. Comparison of the results indicates the superiority of neural networks over that of the MLR method indicating the nonlinear behaviors of the ECD system. Inspection of the models reveals that the surface of the molecules play different roles in response factors of two subsets due to rotation of one phenyl group with respect to the other for the subset consisting of larger number of chlorine atoms. PMID- 14753692 TI - Simultaneous determination of semivolatile organic compounds in indoor air by gas chromatography-mass spectrometry after solid-phase extraction. AB - A method is described for simultaneous determination of semivolatile organic compounds (SVOCs) in indoor air by gas chromatography-mass spectrometry (GC-MS). The selected 73 SVOCs were collected using combined adsorbents (quartz fiber filter disk and Empore disk) for 24 h at a 5.0 l/min flow rate. The SVOCs collected were extracted with acetone, concentrated, then analyzed by an internal standard method. Forty compounds (19 plasticizers and flame retardants; 19 insecticides; 1 synergist; and 1 fungicide) among the target SVOCs were determined accurately and precisely. The method of detection limits for these compounds were approximately 0.5 ng/m3 for most of the SVOCs. The collected SVOC samples could be stored for up to 1 month at 4 C in the refrigerator. PMID- 14753693 TI - Direct thermal extraction and gas chromatographic-mass spectrometric determination of volatile compounds of extra-virgin olive oils. AB - The instrumental performances of a Thermo Desorption-Cooled Injection System coupled with a gas chromatography-mass spectrometer (GC-MS) were improved by a Plackett-Burman experimental design for the direct thermal extraction of volatile compounds from extra-virgin olive oils. The obtained experimental conditions were applied to the analysis of samples from West Liguria (cv. Taggiasca > or = 90%) and Spain (cv. Arbequina), which shared such similar sensorial features that Taste Panel did not distinguish them. Principal component analysis (PCA) was then applied to the experimental data. Three linear combinations of the amounts of the lipoxygenase oxidation products proved to be decisive and sufficient in the differentiation of the two groups of samples. PMID- 14753694 TI - Characterization of archaeological frankincense by gas chromatography-mass spectrometry. AB - A simple gas chromatography-mass spectrometry (GC-MS) method has been developed for the characterization of frankincense in archaeological samples. After trimethylsilylation of the methanolic extract, 15 triterpenoids have been found among the chemical constituents of commercial olibanum (alpha-boswellic acid, 3-O acetyl-alpha-boswellic acid, beta-boswellic acid, 3-O-acetyl-beta-boswellic acid, alpha-amyrin, beta-amyrin, lupeol, 3-epi-beta-amyrin, 3-epi-beta-amyrin, 3-epi lupeol, alpha-amyrenone, beta-amyrenone, lupenone, 3alpha-hydroxy-lup-20(29)-en 24-oic acid and 3-O-acetyl-hydroxy-lup-20(29)-en-24-oic acid). These compounds have been unequivocally identified by retention time and mass spectral comparison with pure standards previously isolated, for the most part, in our laboratory. Within these triterpenes, acid ones, the corresponding O-acetates, and their products of degradation were found to be characteristic of frankincense (Boswellia resin). The presence of these unusual triterpenic compounds in an archaeological resinous sample, recovered during excavations from Dahshour site (Egypt, XIIth Dynasty), enabled us to identify unambiguously frankincense resin among several other materials. Additional chromatographic peaks of this sample were assigned to broad chemical classes using retention time and mass spectra features. PMID- 14753695 TI - Study of the influence of surfactants on the transfer of gases into liquids by inverse gas chromatography. AB - The experimental technique of the reversed-flow version of inverse gas chromatography was applied for the study of effects of surfactants in reducing air-water exchange rates. The vinyl chloride (VC)-water system was used as a model, which is of great importance in environmental chemistry. Using suitable mathematical analysis, various physicochemical quantities were calculated, among which the most significant are: Partition coefficients of the VC gas between the surfactant interface and the carrier gas nitrogen, as well as between the bulk of the water + surfactant solution and the carrier gas nitrogen, overall mass transfer coefficients of VC in the liquid (water + surfactant) and the gas (nitrogen) phases, water and surfactant film transfer coefficients, nitrogen, water and surfactant phase resistances for the transfer of VC into the water solution, relative resistance of surfactant in the transfer of VC into the bulk of solution, exchange velocity of VC between nitrogen and the liquid solution, and finally the thickness of the surfactant stagnant film in the liquid phase, according to the three phase resistance model. From the variation of the above parameters with the surfactant's concentration, important conclusions concerning the effects of surfactants on the transfer of a gas at the air-liquid interface, as well as to the bulk of the liquid were extracted. An interesting finding of this work was also that by successive addition of surfactant, the critical micelle concentration of surfactant was obtained, after which follows a steady state for the transfer of the gas into the water body, which could be attributed to the transition from mono- to multi-layer state. PMID- 14753696 TI - Properties of poly-aminophenylboronate coatings in capillary electrophoresis for the selective separation of diastereoisomers and glycoproteins. AB - The polymerisation of 3-aminophenylboronic acid (APBA) in aqueous environment has been used for the open tubular modification of capillary electrophoresis (CE) capillaries. Being poly-APBA endowed with boronic acid, aromatic rings and secondary amines groups, it posses a variety of functional groups affecting selectivity. Diastereoisomers (e.g. ascorbic and isoascorbic acid) and proteins (e.g. haemoglobins) were successfully separated onto poly-APBA column, by means of a combination of electrophoresis and open tubular electrochromatography. The mechanism of selection was investigated: results indicate an interplay between enhancing or silencing the contribution of the protonable functionalities (amino groups, boronic acid). The properties of APBA polymer coating make it attractive for CE separation and for further application in affinity separations and chip technologies. PMID- 14753697 TI - Dynamic extraction in rotating coiled columns, a new approach to direct recovery of polycyclic aromatic hydrocarbons from soils. AB - A new approach to the direct recovery of polycyclic aromatic hydrocarbons (PAHs) from environmental solid samples has been proposed. It has been shown that rotating coiled columns (RCCs) earlier used mainly in countercurrent chromatography can be successfully applied to the fast continuous-flow extraction of PAHs from soils. A particulate solid sample was retained in the rotating column as the stationary phase under the action of centrifugal forces while a mixture of organic solvents (acetone-cyclohexane, 1:1, v/v) was continuously pumped through. The separation procedure requires less than half an hour, complete automation being possible. No clean-up step is needed before the subsequent HPLC- analysis of extracts. Besides, the dynamic multistage extraction performed in the rotating column at room temperature and normal pressure may have nearly the same efficiency as accelerated batch solvent extraction repeated three times at 150 degrees C and 14 MPa. Contents of PAHs in extracts obtained by using both methods are in good agreement with the certified data on the PAHs concentrations in the soil samples. The use of appropriate "mild" solvents for the dynamic extraction in rotating columns may be very perspective for the simulation of naturally occurring processes and determination of environmentally relevant forms of PAHs and other pollutants in environmental solids. A particular emphasis could be placed on time-resolved (kinetic) studies of the mobilization of toxicants in soil systems. PMID- 14753698 TI - Preparative isolation and purification of isoflavan and pterocarpan glycosides from Astragalus membranaceus Bge. var. mongholicus (Bge.) Hsiao by high-speed counter-current chromatography. AB - (3R)-(-)-7,2'-Dihydroxy-3',4'-dimethyl isoflavan-7-O-beta-D-glucopyranoside and (6aR, 11aR) 9,10-di-methoxypterocarpan-3-O-beta-D-glucopyranoside were separated from the ethyl acetate extract of the root of Astragalus membranaceus Bge. var. mongholicus (Bge.) Hsiao by high-speed counter-current chromatography (HSCCC). A two-phase system composed of ethyl acetate-ethanol-acetic acid-water (4:1:0.25:5, v/v) was selected by analytical HSCCC. Preparative HSCCC yielded, from 100 mg of the partially purified extract, 50 mg of isoflavan glycoside and 10 mg of pterocarpan glycoside each at over 95% purity by high-performance liquid chromatography (HPLC) analysis. Their structures were identified by MS, 1H NMR and 13C NMR. PMID- 14753699 TI - Comparison of protein precipitation methods for sample preparation prior to proteomic analysis. AB - Protein samples should be free of salt and other disturbing agents and have an appropriate concentration to be suitable for two-dimensional (2D) electrophoresis, the principal step of proteomics. To find the most efficient method for sample preparation, we used human plasma and compared four widely applied precipitation methods, using trichloroacetic acid (TCA), acetone, chloroform/methanol and ammonium sulfate, as well as ultrafiltration. Precipitation with TCA and acetone and ultrafiltration resulted in an efficient sample concentration and desalting. We also found that ammonium sulfate fractionation can efficiently remove albumin, which represents more than 50% of plasma proteins. PMID- 14753701 TI - Improved model of multicomponent adsorption in reversed-phase liquid chromatography. AB - This work presents a modification to the real adsorbed solution model using a Flory-Huggins type of expression that was reported previously. This modification consists in replacing the Flory-Huggins activity coefficient by the spreading pressure dependent approach. This new model takes into account explicitly the adsorbate-adsorbate interactions taking place in the adsorbed phase. It provides an excellent prediction of the competitive, ternary adsorption equilibrium of benzyl alcohol, 2-phenylethanol and 2-methyl benzyl alcohol observed in a reversed-phase liquid chromatographic system using information merely derived from the single-component adsorption experimental data. PMID- 14753700 TI - Solid-phase microextraction with on-fiber derivatization for the analysis of anti inflammatory drugs in water samples. AB - A sensitive and solvent-free procedure for the determination of non-steroidal acidic anti-inflammatory drugs in water samples was optimized using solid-phase microextraction (SPME) followed by on-fiber silylation of the acidic compounds and gas chromatography-mass spectrometry (GC-MS) determination. Microextraction was carried out directly over the filtered water samples using a polyacrylate fiber. Derivatization was performed placing the SPME fiber, loaded with the extracted analytes, in the headspace of a vial containing 50 microl of N-methyl-N (tert-butyldimethylsilyl)-trifluoroacetamide (MTBSTFA). Derivatives were desorbed for 3 min in the GC injector. Influence of several parameters in the efficiency of microextraction (volume of sample, time, pH, type of fiber coating, etc.) and derivatization steps (time, temperature and volume of MTBSTFA) was systematically investigated. In the optimal conditions an excellent linearity over three orders of magnitude and quantification limits at the ng/l level (from 12 to 40 ng/l) were achieved. The proposed method was applied to the determination of acidic compounds in sewage water and results compared to those obtained using solid phase extraction (SPE) followed by the derivatization of the compounds in the organic extract of the solid-phase extraction cartridge. PMID- 14753702 TI - General model for the achiral chromatography of enantiomers forming dimers: application to binaphthol. AB - This works deals with the development of a model for the description of the chromatographic behavior of enantiomers forming dimers on achiral stationary phases. The model describes the formation of both homochiral and heterochiral dimers by introducing two equilibrium constants, K(homo) and K(hetero), which have to be independently estimated. The adsorption of both monomeric and dimeric species is described by a bi-Langmuir isotherm. Both adsorption and dimerization equilibrium models are implemented in a standard equilibrium-dispersive model of the chromatographic column. The isotherm parameters are determined by fitting of pulses constituted either of the pure enantiomers or of non racemic mixtures of both enantiomers. The validity of the model is then assessed by application to the binaphthol case. PMID- 14753703 TI - Repeatability and reproducibility of high-concentration data in reversed-phase liquid chromatography. III. Isotherm reproducibility on Kromasil C18. AB - Single component equilibrium isotherms of six compounds (aniline, caffeine, ethylbenzene, phenol, propranolol, and theophylline) were determined by the inverse method on 10 Kromasil-C18 columns, using water-methanol solutions as the mobile phase. This method offers an economic and fast isotherm determination by means of the overloaded band profiles of the compounds. Five out of the ten columns used in this test come from the same batch whilst the other five columns represent five additional batches. Statistical evaluation was used to assess the reproducibility of the isotherm parameters. We found that the column-to-column reproducibility of the isotherm parameters is of the same magnitude as the batch to-batch reproducibility (with the exception of one outlier column). In most of the cases, the reproducibilities of the saturation capacities and that of the retention factors are excellent, they are typically between 1.2 and 3%, and very often below 2%. Within the limits of the experimental precision, these results agree with those obtained earlier, using a conventional method of isotherm determination. PMID- 14753704 TI - 1H nuclear magnetic resonance study of the molecular imprinting of (-)-nicotine: template self-association, a molecular basis for cooperative ligand binding. AB - In the present study, the interactions of components in a (-)-nicotine molecular imprinting polymerization mixture have been studied by 1H NMR spectroscopy. The dissociation constants for complexation of template by a functional monomer analogue, acetic acid, have been determined. Nicotine was shown to self-associate at concentrations comparable to those used in previous molecular imprinting studies (app K(diss) = 0.082M in CDCl3 at 298 K). The extent of self-association was enhanced by the presence of acetic acid. Previous studies on (-)-nicotine imprinted methacrylic acid-ethylene dimethacrylate co-polymers suggested the involvement of recognition sites for template-template complexes. Collectively these results provide the first direct evidence for the presence of template template complexes, and support the previously hypothesized basis for cooperative ligand recognition events in this polymer system. PMID- 14753705 TI - New insights in the spatially resolved dynamic pH measurement in macroscopic large absorbent particles by confocal laser scanning microscopy. AB - Both, experimental investigation of protein adsorption processes and mathematical models describing such processes indicate, that the pH in the absorbent particle might be the key factor for an improved understanding of these chromatographic processes. Thus, a technique aiming at the spatially resolved pH measurement in macroscopic large absorbent particles is presented. The first application of this method, being based on confocal laser scanning microscopy (CLSM), revealed an apparent dependence of the pH calibration curve on the scanning depth. By a model based approach, factors distorting the measurement signal are identified: The wavelength-dependent light scattering and the re-absorption of emitted light. The resulting consequences for further development and application of CLSM based techniques to measure pH in macroscopic large absorbent particles are illustrated and discussed. PMID- 14753706 TI - Liquid chromatographic separation of the enantiomers of trans-chlordane, cis chlordane, heptachlor, heptachlor epoxide and alpha-hexachlorocyclohexane with application to small-scale preparative separation. AB - Analytical high-performance liquid chromatographic separations of the individual enantiomers of five polychlorinated compounds were obtained on polysaccharide stereoselective HPLC columns. The enantiomers of the pesticides trans-chlordane, cis-chlordane and heptachlor were separated on CHIRALCEL OD using a hexane mobile phase. The enantiomers of the heptachlor metabolite, heptachlor epoxide, were separated on CHIRALPAK AD using a methanol mobile phase. The enantiomers of alpha hexachlorocyclohexane (alpha-HCH), were separated on CHIRALCEL OJ using a hexane/2-propanol mobile phase. Similar chromatographic conditions using preparative columns were used to isolate approximately 250 mg of each of the individual enantiomers. The purified individual enantiomers have been submitted for testing of their endocrine disruptor (ED) activity. PMID- 14753707 TI - Liquid chromatography--multiple tandem mass spectrometry for the determination of ten azaspiracids, including hydroxyl analogues in shellfish. AB - Azaspiracids (AZAs) are a group of polyether toxins that cause food poisoning in humans. These toxins, produced by marine dinoflagellates, accumulate in filter feeding shellfish, especially mussels. Sensitive liquid chromatography electrospray ionisation mass spectrometry (LC-ESI-MS(n)) methods have been developed for the determination of the major AZAs and their hydroxyl analogues. These methods, utilising both chromatographic and mass resolution, were applied for the determination of 10 AZAs in mussels (Mytilus edulis). An optimised isocratic reversed phase method (3 microm Luna-2 C18 column) separated 10 azaspiracids using acetonitrile/water (46:54, v/v) containing 0.05% trifluoroacetic acid (TFA) and 0.004% ammonium acetate in 55 min. Analyte determination using MS3 involved trapping and fragmentation of the [M + H]+ and [M + H - H2O]+ ions with detection of the [M + H - 2H2O]+ ion for each AZA. Linear calibrations were obtained for AZA1, using spiked shellfish extracts, in the range 0.05-1.00 microg/ml (r2 = 0.997) with a detection limit of 5 pg (signal : noise = 3). The major fragmentation pathways in hydroxylated azaspiracids were elucidated using hydrogen/deuterium (H/D) exchange experiments. An LC-MS3 method was developed using unique parent ions and product ions, [M + H - H2O - CgH10O2R1R3]+, that involved fragmentation of the A-ring. This facilitated the discrimination between 10 azapiracids, AZA1-10. Thus, this rapid LC-MS3 method did not require complete chromatographic resolution and the run-time of 7 min had detection limits better than 20 pg for each toxin. PMID- 14753708 TI - Effect of the detergent Tween-20 on the DNA affinity chromatography of Gal4, C/EBPalpha, and lac repressor with observations on column regeneration. AB - C/EBPalpha, Gal4, and lac repressor, representing three different transcription factor homology families, were expressed as fusion proteins and used to characterize the effects of column aging, Mg2+, the nonionic detergent Tween-20, column loading, and bovine serum albumin on DNA-affinity chromatography. When lac repressor-beta-galactosidase fusion protein is loaded onto a new DNA-Sepharose column, less elutes from a new column than one that has been used two or more times. Higher amounts of lac repressor, the Green Fluorescent Protein fusions with CAAT enhancer binding protein (C/EBPalpha) and Gal4, elute from the columns when 0.1% Tween-20 is added to the mobile phase. The amount of improvement found depends upon the transcription factor studied and the amount of the protein loaded on the column; lac repressor and Gal4 are eluted in higher amounts over a large range of protein loads while C/EBP shows the greatest effect at low protein loads. This detergent effect is seen when either Sepharose or silica is used for the stationary phase. Including bovine serum albumin in the mobile phase gives a similar though lesser improvement to that observed with Tween-20. Mg2+ or EDTA in the mobile phase gave similar chromatography for C/EBP; since EDTA protects columns from DNases, its inclusion in the mobile phase is preferred. After extended use, the DNA affinity columns no longer bind transcription factors and this is not due to losses of DNA from the columns. Two simple methods (sodium dodecylsulfate and KSCN) were developed to regenerate such worn out columns. PMID- 14753709 TI - Factorial screening of antibody purification processes using three chromatography steps without protein A. AB - Protein A affinity chromatography is often employed as a capture step to meet the purity, yield, and throughput requirements for pharmaceutical antibody purification. However, a trade-off exists between step performance and price. Protein A resin removes 99.9% of feed stream impurities; however, its price is significantly greater than those of non-affinity media. With many therapeutic indications for antibodies requiring high doses and/or chronic administration, the consideration of process economics is critical. We have systematically evaluated the purification performance of cation-exchange, anion-exchange, hydroxyapatite, hydrophobic interaction, hydrophobic charge induction, and small molecule ligand resins in each step of a three-step chromatographic purification process for a CHO-derived monoclonal antibody. Host cell proteins were removed to less-than-detectable for three processes (cation-exchange-anion-exchange hydrophobic interaction chromatography, cation-exchange-anion-exchange-mixed cation-exchange chromatography, and cation-exchange-mixed cation-exchange-anion exchange chromatography). The order of the process steps affected purification performance significantly. PMID- 14753710 TI - Quantitation of SU1 1248, an oral multi-target tyrosine kinase inhibitor, and its metabolite in monkey tissues by liquid chromatograph with tandem mass spectrometry following semi-automated liquid-liquid extraction. AB - SU11248 is a potent inhibitor of PDGFR, VEGFR, KIT, and Flt3, and is currently under Phase I clinical evaluation as an anticancer drug. A sensitive and specific analytical method for the quantitation of SU11248 and its metabolite in several monkey tissues (liver, kidney, brain and white fat) using LC-MS-MS following semi automated liquid-liquid extraction (LLE) was developed and validated. Amounts of 50 mg of tissue were homogenized using an ultrasonic processor. After addition of the stable labelled internal standard (IS) and ammonium hydroxide (0.3%), samples were extracted with 2.5 ml of tert-butyl methyl ether. Following centrifugation, aliquots of 1.8 ml of the organic phase were transferred into a 96-well plate. The Packard Multiprobe II robotic liquid handler was used to perform all steps mentioned above. The organic phase was dried and the residue was reconstituted with 800 microl of 15 mM ammonium formate buffer solution (pH 3.25) using a Tomtec Quadra 96 workstation. Aliquots of 10 microl of the resulting solution were injected into the LC-MS-MS system. A Symmetry Shield C8 column (50 mm x 2.1 mm, 3.5 microm) was used to perform the chromatographic analysis. The mobile phase was 15 mM ammonium formate buffer solution (pH 3.25)-acetonitrile (74:26 (v/v)) with a flow-rate of 0.35 ml/min. Retention times of the metabolite and SU11248 were about 2.5 and 3.5 min, respectively. Total cycle time was 5 min. MS detection used the Applied Biosystems-MDS Sciex API 3000 with TurbolonSpray interface and multiple reaction monitoring (MRM) operated in positive ion mode. The method was validated for both compounds over the calibration range of about 2 and 2000 ng/g. The suitability and robustness of the method for in vivo samples were confirmed by analysis of monkey tissues from animals dosed with SU11248. PMID- 14753711 TI - Extraction of plasmid DNA from Escherichia coli cell lysate in a thermoseparating aqueous two-phase system. AB - The primary purification of a 6.1 kilo base pair (kbp) plasmid from a desalted alkaline lysate has been accomplished by a thermoseparating aqueous two-phase system [(50% ethylene oxide-50% propylene oxide)-Dextran T 500]. The partitioning of the different nucleic acids (plasmid DNA, RNA, genomic DNA) in the thermoseparating aqueous two-phase system was followed both qualitatively by agarose gel electrophoresis and quantitatively by analytical chromatography (size exclusion- and anion-exchange mode) and PicoGreen fluorescence analysis. The experimental results showed a complete recovery of the plasmid DNA to the top phase, while 80% of total RNA and 58% of total protein was discarded to the bottom phase. Moreover, a 3.8-fold volume reduction of the plasmid DNA solution was achieved. By using a final thermoseparating step, the EO50PO50 polymer could be efficiently recycled, resulting in plasmid solution containing less than 1% polymer. The developed thermoseparating aqueous two-phase system shows great potential for the large-scale processing of plasmid DNA. PMID- 14753712 TI - Simultaneous determination of twelve inorganic and organic arsenic compounds by liquid chromatography-ultraviolet irradiation-hydride generation atomic fluorescence spectrometry. AB - A coupling between column liquid chromatography (LC) and atomic fluorescence spectrometry was developed for arsenic speciation. After separation, the compounds are oxidised on-line by UV irradiation, volatilised by hydride generation and carried to the detector by a stream of argon. A combination of anion-exchange and hydrophobic interactions in a single column (Dionex AS7) was found suitable for the simultaneous separation of organic and inorganic species. Twelve compounds (arsenite, arsenate, monomethylarsonic acid, dimethylarsinic acid, arsenobetaine, arsenocholine, trimethylarsine oxide, tetramethylarsonium ion and four arsenosugars) were separated using an acetate buffer and a nitric acid solution as mobile phases. Limits of detection are 4-22 pg. The technique was applied to three marine samples. Arsenobetaine was detected as the main species in all samples, with concentrations varying from 59 to 1947 ng(As) g(-1) of fresh mass. PMID- 14753713 TI - Development and validation of a liquid chromatographic method for determination of lacidipine residues on surfaces in the manufacture of pharmaceuticals. AB - A high-performance liquid chromatographic (HPLC) method for the assay of lacidipine residues in swabs collected from various surfaces involved in drug manufacture is described. The swabbing procedure using two cotton swabs was validated applying a wipe test. An RP-HPLC method, developed to determine low quantities of the drug in the presence of its main impurities, was also validated. To remove drug residues from stainless steel and glass surfaces, the first cotton swab must be soaked preferably in acetonitrile whereas, on vinyl surfaces better results are obtained using methanol. The HPLC method selected involves a C12 column, at 40 degrees C, a mixture of acetonitrile-0.05 M ammonium acetate (88:12, v/v) as a mobile phase and UV detection at 282 nm. Recoveries obtained are strongly dependent on the type of surface tested, being higher on stainless steel. The surface material has also different influence on the drug stability. The method was validated over a range of 0.5-100 microg/400 cm2 and had a detection limit of 0.1 microg/400 cm2. PMID- 14753714 TI - Determination of chlorophyll in plant samples by liquid chromatography using zinc phthalocyanine as an internal standard. AB - Chlorophyll analysis at high precision and accuracy is limited by the lack of suitable, commercially available internal standards for HPLC analysis. Here, the commercially available dye zinc-phthalocyanine is presented as a new internal standard to quantify chlorophylls in vegetable foods and to detect chlorophyll degradation products. The technique was applied to chlorophyll analysis of a selection of vegetable foods. Pigments were extracted with N,N-dimethylformamide from the vegetables and purified by solid phase extraction. Chlorophyll a, a', b, b', corresponding pheophytins, and zinc-phthalocyanine were separated by HPLC using a C18 reverse-phase column and fluorescence detection. PMID- 14753715 TI - Retention of Cr(III) by high-performance chelation ion chromatography interfaced to inductively-coupled plasma mass spectrometric detection with collision cell. AB - High-performance chelation ion chromatography (HPCIC) was employed to retain cationic Cr(III) on an anion-exchange column and hence allow the separation of the two most prevalent forms of chromium, Cr(II) and Cr(VI). A mobile phase of nitric acid was utilized at pH = 1.5; additionally, 2,6-pyridinedicarboxylic acid was used at a concentration of 6 mM. Additives with different structural characteristics were used in an effort to elucidate retention mechanisms. Inductively-coupled plasma mass spectrometry was used for chromium detection. A collision cell was utilized to reduce chloride-based polyatomic ions that may interfere with the detection of Cr(III), and a detection limit study yielded levels in the low part-per-billion range. The newly developed method was applied to the chromatographic analysis of samples of an incubation medium containing Cr(VI) incubated with cell nuclei. PMID- 14753716 TI - Analysis of volatile components of Lavandula luisieri L. by direct thermal desorption-gas chromatography-mass spectrometry. AB - Volatile components of wild samples of Lavandula luisieri collected in Central and Southern Spain have been analyzed by direct thermal desorption coupled to gas chromatography-mass spectrometry (DTD-GC-MS). This method requires only 10-20 mg of dry sample, allowing to obtain qualitative and quantitative results from different plant parts such as flowers and leaves. Average volatile yield calculated from 51 individual plants was higher for leaves (9.7mg g(-1)) than for flowers (2.9mg g(-1)). Samples presented a high variation in their yield and composition. Major components were camphor and 1,8-cineole (up to 80.9 and 76.7% in leaves; 87.8 and 85.2% in flowers, respectively); however, these compounds were not detected in several samples. Other major component (up to 60% in flowers and leaves) was 2,3,5,5-tetramethyl-4-methylene-2-cyclopenten-1-one. Multivariate analysis was applied to quantitative data from nine selected compounds in order to show the presence of several patterns in plant composition which were only partially related to the site of collection. PMID- 14753717 TI - Quality assurance of commercial beeswax. Part I. Gas chromatography-electron impact ionization mass spectrometry of hydrocarbons and monoesters. AB - The use of low-temperature capillary gas chromatography coupled to electron impact mass spectrometry for the characterization of crude beeswaxes yielded by Apis mellifera is described. The system allows the identification of a great number of compounds, some of them not reported till now in beeswax, such as a family of ethyl esters, tetracosyl oleate, and several saturated and unsaturated hydrocarbons. The information acquired makes possible the differentiation between pure beeswax and some foundation beeswax samples where mixture of pure beeswax with another substances is suspected. PMID- 14753718 TI - Optimization of a microwave-assisted derivatization-extraction procedure for the determination of chlorophenols in ash samples. AB - A procedure for the determination of 17 chlorophenolic compounds in ash samples obtained from the incineration of waste materials is described. Analytes were simultaneously derivatized with acetic anhydride in presence of triethylamine (TEA), and extracted from the sample in a mixture of n-hexane acetone using a microwave system equipped with closed extraction vessels. Influence of five experimental parameters (volume of TEA and acetic anhydride, extraction time and temperature, as well as the volume of n-hexane acetone) on the yield of the derivatization-extraction procedure was systematically studied using a uniform experimental design at four levels, followed by a conventional factorial design at two levels. Under optimal extraction conditions, recoveries from 72 to 94% were obtained for a spiked ash sample with a carbon content of 8.7%. Quantification limits of the proposed procedure ranged from 2 to 5 ng/g using GC MS as detection technique. The proposed method was applied to the determination of chlorophenols in three ash samples obtained from different incineration plants. Total chlorophenol contents of 423 and 135 ng/g were found in two of these samples. PMID- 14753719 TI - Characterization and determination of fatty acids in fish oil using gas chromatography-mass spectrometry coupled with chemometric resolution techniques. AB - Characterization and determination of a complex mixture of fatty acid methyl esters was performed for commercial fish oil using two-dimensional GC-MS data coupled with resolution techniques. Various principle component analysis methods such as significant factor analysis and fixed size moving window evolving factor analysis were used for the number of factors, zero concentration and selective regions. Then, the convoluted chromatograms were resolved into pure chromatograms and mass spectra using heuristic evolving latent projections (HELP) method. Fatty acids of C16:1omega7, C18:4omega3, C18:1omega11, C18:1omega9, C18:0, C20:2omega6, C20:1omega9, C22:1omega11, C22:1omega9 and C24:1omega9 were resolved an fied by using similarity searches between deconvoluted mass spectra and MS database, in different parts of total ion current chromatogram. Window target testing factor analysis is also applied for confirming the presence or absence of target analytes. The results of the present work show that combination of hyphenated chromatographic methods and resolution techniques provide a complementary method for accurate analysis of real multi-component systems such as fish oil. PMID- 14753720 TI - Determination of natural and synthetic estrogens in water by gas chromatography with mass spectrometric detection. AB - A procedure for the determination of six natural and synthetic estrogens (diethylstilbestrol, estrone, 17beta-estradiol, mestranol, 17alpha ethinylestradiol and estriol) in water samples is described. Samples, up to 2000 ml, were concentrated using Oasis HLB solid-phase extraction cartridges. Analytes were derivatized with N-methyl-N-(trimethylsilyl)trifluoroacetamide and determined by GC-MS or GC-MS-MS. The reactivity of several silylation reagents versus aliphatic and aromatic hydroxyl groups contained in the structure of the selected analytes was evaluated. Influence of parameters such as sample pH, nature of the water samples and derivatization conditions on the performance of the whole analytical procedure was systematically studied. Under optimal conditions, quantification limits between 1 and 3 ng/l were achieved for the determination of the considered estrogens in sewage water. PMID- 14753721 TI - Identification of diterpenes in canvas painting varnishes by gas chromatography mass spectrometry with combined derivatisation. AB - A derivatisation method that combines the formation of ethyl esters from the carboxylic groups and trimethylsilyl ethers from hydroxyl groups of the components of diterpenic resins is presented in this paper. This methodology involves two experimental steps: (1) formation of ethyl esters using ethyl chloroformate; and (2) the esterified compounds are lead to react with trimethylsilylimidazole to form the corresponding trimethylsilyl ethers. The main advantage of the proposed method is the possibility of performing simultaneously the analysis of amino acids from proteins, fatty acids from drying oils, and diterpenic compounds from natural resins usually found in works of art. This methodology is of considerable interest due to the requirements of minimum sampling that usually involves the analysis of works of art. A chemometric study has been developed to adjust the optimal working conditions of the proposed derivatisation method in which chromatographic peak areas of the larixyl acetate derivative and the abietic acid derivative referred to n-hexadecane as internal standard have been compared. Samples of Venetian turpentine naturally aged have been used in this study. Finally, the efficiency of the proposed derivatisation method has been tested on other diterpenic resins and pigments commonly used in fine arts such as Strasbourg turpentine, Canada balsam, colophony, copper resinate and a sample from a Renaissance Altarpiece. PMID- 14753722 TI - Direct resistively heated column gas chromatography (Ultrafast module-GC) for high-speed analysis of essential oils of differing complexities. AB - This study applies Ultrafast module-GC (UFM-GC) with direct resistively heated columns to routine analysis of a group of essential oils of differing complexities (chamomile, peppermint, rosemary and sage). Essential oils were analysed by conventional GC with conventional inner diameter (i.d.) columns (0.25 mm) of different lengths (5 and 25 m long) and by Fast GC and Ultrafast module-GC with narrow bore columns (0.1 mm i.d., 5 m long). Column performance were evaluated and compared through their Grob test, separation number and peak capacity. Ultrafast module-GC was successful in the qualitative and quantitative analysis of essential oils of different compositions with analysis times between 40 s and 2 min versus 20-60 min required by conventional GC. Critical pairs or groups of components were separated by carefully tuning selectivity of the stationary phase to compensate for loss of efficiency due to the use of short columns and high temperature rates. The Ultrafast module-GC results of peppermint e.o. analyses were also validated and compared to those obtained by conventional GC; by measuring precision over time (i.e. repeatability and intermediate precision) and accuracy. Ultrafast module-GC showed a good separation reproducibility affording reliable component identification through the relative retention times and quantitative determination through normalised peak areas. Accuracy data also showed that Ultrafast module-GC and conventional GC normalised areas and areas percentage were perfectly comparable. PMID- 14753723 TI - Determination of methylmercury in fish using focused microwave digestion following by Cu2+ addition, sodium tetrapropylborate derivatization, n-heptane extraction, and gas chromatography-mass spectrometry. AB - The analytical procedure for analysis of methylmercury in fish was developed. It involves microwave-assisted digestion with alkaline solution (tetramethylammonium hydroxide), addition of Cu2+, aqueous-phase derivatization of methylmercury with sodium tetrapropylborate, and subsequent extraction with n-heptane. The methylmercury derivative was desorbed in the splitless injection port of a gas chromatograph and subsequently analyzed by electron impact mass spectrometry. Optimum conditions allowed sample throughout to be controlled by the instrumental analysis time (near 7 min per sample) but not by the sample preparation step. At the power of 15-30, 45, and 60-75 W, sample preparation time is only 3.5, 2.5, and 1.5 min, respectively. The proposed method was finally validated by the analysis of three biological certified reference materials, BCR CRM 464 tuna fish, NRC DORM-2 dogfish muscle, and NRC DOLT-2 dogfish liver. The detection limit of the overall procedure was found to be 40 ng/g of biological tissue for methylmercury. The recovery of methylmercury was 91.2-95.3% for tuna, 89.3-94.7% for marlin, and 91.7-94.8% for shark, respectively. The detected and certified values of methylmercury of three biological certified reference materials were as follows: 5.34 +/- 0.30 microg/g (mean +/- S.D.) and 5.50 +/- 0.17 microg/g for CRM 464 tuna fish, 4.34 +/- 0.24 and 4.47 +/- 0.32 microg/g for NRC DORM-2 dogfish muscle, and 0.652 +/- 0.053 and 0.693 +/- 0.055 microg/g for NRC DOLT-2 dogfish liver, respectively. It indicated that the method was well available to quantify the methylmercury in fish. PMID- 14753724 TI - Automated headspace solid-phase dynamic extraction to analyse the volatile fraction of food matrices. AB - High concentration capacity headspace techniques (headspace solid-phase microextraction (HS-SPME) and headspace sorptive extraction (HSSE)) are a bridge between static and dynamic headspace, since they give high concentration factors as does dynamic headspace (D-HS), and are as easy to apply and as reproducible as static headspace (S-HS). In 2000, Chromtech (Idstein, Germany) introduced an inside-needle technique for vapour and liquid sampling, solid-phase dynamic extraction (SPDE), also known as "the magic needle". In SPDE, analytes are concentrated on a 50 microm film of polydimethylsiloxane (PDMS) and activated carbon (10%) coated onto the inside wall of the stainless steel needle (5 cm) of a 2.5 ml gas tight syringe. When SPDE is used for headspace sampling (HS-SPDE), a fixed volume of the headspace of the sample under investigation is sucked up an appropriate number of times with the gas tight syringe and an analyte amount suitable for a reliable GC or GC-MS analysis accumulates in the polymer coating the needle wall. This article describes the preliminary results of both a study on the optimisation of sampling parameters conditioning HS-SPDE recovery, through the analysis of a standard mixture of highly volatile compounds (beta-pinene, isoamyl acetate and linalool) and of the HS-SPDE-GC-MS analyses of aromatic plants and food matrices. This study shows that HS-SPDE is a successful technique for HS-sampling with high concentration capability, good repeatability and intermediate precision, also when it is compared to HS-SPME. PMID- 14753725 TI - Thin-layer chromatography of pitch and a petroleum vacuum residue. Relation between mobility and molecular size shown by size-exclusion chromatography. AB - A coal tar pitch and a petroleum vacuum residue have been separated by TLC using pyridine, acetonitrile, toluene and pentane to develop the chromatograms. The bands of material detected were recovered in 1-methyl-2-pyrrolidinone (NMP) solvent and examined by size-exclusion chromatography (SEC) in NMP eluent. The relation between elution time in SEC and mobility on the TLC plate indicated that molecular size increased steadily with increasing immobility on the plate. This relation was reinforced by UV fluorescence spectroscopy in that the fluorescence moved to longer wavelengths with increasing immobility. The molecular size of the material excluded from the porosity of the SEC column remains undefined; some excluded material was found in all of the fractions from both samples. The valley of zero intensity separating the retained material from the excluded material may suggest a change of structure from near-planar in the retained region to three dimensional in the excluded region. PMID- 14753726 TI - Analysis of drying oils used as binding media for objects of art by capillary electrophoresis with indirect UV and conductivity detection. AB - Capillary electrophoresis (CE) was applied to analyse the long-chain fatty acid composition of vegetable oils, and their degradation products formed upon ageing when drying oils are used as binding media. The analytes were detected with contactless conductivity detection (CCD) and indirect UV absorption, both detectors positioned on-line at the separation capillary. The long-chain fatty acids were resolved in a background electrolyte (BGE) consisting of phosphate buffer (pH = 6.86, 15 mM) containing 4 mM sodium dodecylbenzensulfonate, 10 mM Brij 35, 2% (v/v) 1-octanol and 45% (v/v) acetonitrile. As in this system dicarboxylic analytes, the products of oxidative degradation of unsaturated fatty acids, cannot be determined, a suitable background electrolyte was developed by the aid of computer simulation program PeakMaster. It makes use of a 10 mM salicylic acid, 20 mM histidine buffer, pH 5.85, which combines buffering ability with the optical properties obligatory for indirect UV detection. This buffer avoids system eigenpeaks, which are often impairing the separation efficiency of the system. Separation of the dicarboxylic analytes was further improved by a counter-directed electroosmotic flow (EOF), obtained by dynamically coating the capillary wall with 0.2 mM cetyltrimethylammonium bromide. Long-chain fatty acids and their decomposition products could be determined in recent and aged samples of drying oils, respectively, and in samples taken from two paintings of the 19th century. PMID- 14753727 TI - Applicability of predictive models to the peptide mobility analysis by capillary electrophoresis-electrospray mass spectrometry. AB - The prediction of peptide mobility by capillary electrophoresis (CE) coupled to electrospray mass spectrometry (MS) is studied in order to verify the validity of the semi-empirical models developed in classical CE. This work relies on the experimental determination of the electrophoretic mobilities of 68 peptides, different in charge and in size. The results indicate that the prediction is possible in CE-MS experiments, in spite of the restraints inherent in the coupling conditions. The best fit of experimental data was obtained with the Offord's model. The efficiency of the model was confirmed by the analysis of a peptide mixture in CE-MS. PMID- 14753728 TI - Application of solid-phase extraction and micellar electrokinetic capillary chromatography to the study of hydrolytic and photolytic degradation of phenoxy acid and phenylurea herbicides. AB - A degradation study of two phenoxy acid [(2,4-dichlorophenoxy) propanoic acid and (2,4,5-trichlorophenoxy) acetic acid] and two phenylurea (diuron and monolinuron) herbicides, spiked at 50 ppb in water, was performed. Some samples were subjected to neutral and basic hydrolysis; other samples were subjected to photolysis using either sunlight or a xenon arc lamp. After degradation, the water samples were preconcentrated using solid-phase extraction (SPE) with Carbopack B columns and analysed by a micellar electrokinetic capillary chromatography (MECC) system with UV detection at 210 nm. Phenoxyacetic acids were not degraded neither by hydrolysis nor by sunlight photolysis, but they were photodegraded when they were exposed to a xenon arc lamp, with half-lives around 300 min. Phenylurea herbicides were hydrolysed at the two-tested pH, with half-lives varying from 25 to 290 days. The main hydrolysis products were the corresponding chloroanilines. Diuron and monolinuron were also degraded when they were exposed to sunlight and xenon arc lamp. The main photodegradation pathway for diuron corresponded to dehalogenation, while for monolinuron dealkylation and hydroxylation were also postulated. The toxicity of the studied herbicides and their degradation products was evaluated by means of Microtox tests. The obtained results indicated that the toxicity of the degraded samples was higher than the toxicity of the herbicides. PMID- 14753729 TI - Reversed-phase liquid chromatographic method for the determination of ochratoxin A in wine. AB - In this paper, we propose a new, rapid, highly sensitive and reproducible RP-HPLC FLD method for the detection of ochratoxin A (OTA) in wine, by directly injecting the liquid in the chromatographic system without any extraction or clean-up. An alkaline mobile phase (NH4Cl:CH-CN 85:15 (v/v), 20 mM, pH 9.8) was used to obtain a distinct fluorescence enhancement. This improvement allows to reach, without an immunoaffinity clean-up or concentration, a detection limit of 0.05 ng/ml, which is similar to those commonly obtained after immunoaffinity purification and acidic elution. The method was statistically validated and directly applied to a series of wine samples. PMID- 14753730 TI - Vignettes in osteoporosis: a road map to successful therapeutics. AB - The diagnosis and management of osteoporosis have become increasingly more complex as new drugs enter the marketplace and meta-analyses of randomized trials with "other" agents become more prolific. We describe five common clinical scenarios encountered in the practice of osteoporosis medicine and various road maps that could lead to successful therapy. INTRODUCTION: The diagnosis and treatment of osteoporosis have changed dramatically in the last decade. Advances in diagnostic technologies and a range of newer treatment options have provided the clinician with a wide array of choices for treating this chronic disease. Despite the issuance of several "guidelines" and practice recommendations, there still remains confusion among clinicians about basic approaches to the management of osteoporosis. This paper should be used as a case-based approach to define optimal therapeutic choices. MATERIALS AND METHODS: Five representative cases were selected from two very large clinical practices (Bangor, ME; Pittsburgh, PA). Diagnostic modalities and treatment options used in these cases were selected on an evidence-based analysis of respective clinical trials. Subsequent to narrative choices by two metabolic bone disease specialists (SG and CR), calculation of future fracture risk and selection of potential alternative therapeutic regimens were reviewed and critiqued by an epidemiologist (DB). RESULTS: A narrative about each case and possible management choices for each of the five cases are presented with references to justify selection of the various therapeutic options. Alternatives are considered and discussed based on literature and references through July 2003. The disposition of the individual patient is noted at the end of each case. CONCLUSIONS: A case-based approach to the management of osteoporosis provides a useful interface between guidelines, evidence-based meta-analyses, and clinical practice dilemmas. PMID- 14753731 TI - Oral daily ibandronate prevents bone loss in early postmenopausal women without osteoporosis. AB - Oral daily ibandronate was investigated for the prevention of bone loss in postmenopausal women without osteoporosis (n = 653). BMD at the lumbar spine and hip were significantly increased (3.1% and 1.8%, respectively; p < or = 0.0001 versus placebo) with 2.5 mg ibandronate after 24 months. Oral ibandronate is a promising option for the prevention of postmenopausal bone loss. INTRODUCTION: Further strategies to manage patients most at risk from developing postmenopausal osteoporosis are required. The objectives of this multicenter, double-blind, randomized, placebo-controlled study were to examine the efficacy, tolerability, and optimal dose of oral daily ibandronate in the prevention of bone loss in postmenopausal women. MATERIALS AND METHODS: In total, 653 women (mean bone mineral density [BMD] T-score > -2.5 at the lumbar spine), who had been postmenopausal for at least 1 year, were allocated to one of four strata based on time since menopause and baseline lumbar spine BMD. Women were randomized to receive calcium (500 mg daily) plus either placebo (n = 162) or ibandronate 0.5 mg (n = 162), 1 mg (n = 166), or 2.5 mg (n = 163) as once-daily oral treatment for 2 years. The primary endpoint was the mean percent change in lumbar spine BMD with ibandronate versus placebo. RESULTS AND CONCLUSIONS: After 2 years, oral daily ibandronate produced a dose-related and sustained maintenance or increase in BMD at the lumbar spine and hip (total hip, femoral neck, trochanter), together with a dose-related reduction in the rate of bone turnover. The greatest nominal increases in spinal and hip BMD were observed with the 2.5-mg dose, which produced statistically significant BMD gains compared with placebo at 6 months and all subsequent time-points at the spine and hip (3.1% and 1.8% increase in lumbar spine and total hip BMD, respectively, versus placebo; p < or = 0.0001 after 24 months). Oral daily ibandronate was well tolerated with an incidence of upper gastrointestinal adverse events similar to placebo. No safety concerns were identified. In summary, oral daily ibandronate 2.5 mg decreases bone turnover, preserves or increases BMD in the spine and proximal femur, and is well tolerated. Oral ibandronate provides a promising option for the prevention of bone loss in postmenopausal women. PMID- 14753732 TI - Beta-adrenergic blockers reduce the risk of fracture partly by increasing bone mineral density: Geelong Osteoporosis Study. AB - This population-based study documented beta-blocker use in 59/569 cases with incident fracture and 112/775 controls. OR for fracture associated with beta blocker use was 0.68 (95%CI, 0.49-0.96). Beta-blockers were associated with higher BMD at the total hip (2.5%) and UD forearm (3.6%) after adjusting for age, anthropometry, and thiazide use. Beta-blocker use is associated with reduced fracture risk and higher BMD. INTRODUCTION: Animal data suggests that bone formation is under beta-adrenergic control and that beta-blockers stimulate bone formation and/or inhibit bone resorption. MATERIALS AND METHODS: We evaluated the association between beta-blocker use, bone mineral density (BMD), and fracture risk in a population-based study in Geelong, a southeastern Australian city with a single teaching hospital and two radiological centers providing complete fracture ascertainment for the region. Beta-blocker use was documented for 569 women with radiologically confirmed incident fractures and 775 controls without incident fracture. Medication use and lifestyle factors were documented by questionnaire. RESULTS: Odds ratio for fracture associated with beta-blocker use was 0.68 (95% CI, 0.49-0.96) for any fracture. Adjusting for age, weight, medications, and lifestyle factors had little effect on the odds ratio. Beta blocker use was associated with a higher BMD at the total hip (2.5%, p = 0.03) and ultradistal forearm (3.6%, p = 0.04) after adjustment for age, anthropometry, and thiazide use. CONCLUSION: Beta-blockers are associated with a reduction in fracture risk and higher BMD. PMID- 14753733 TI - Fracture risk in monoclonal gammopathy of undetermined significance. AB - To assess fractures in monoclonal gammopathy of undetermined significance (MGUS), the precursor of multiple myeloma, we followed 488 Olmsted County, MN, residents with MGUS in a retrospective cohort study. There was a 2.7-fold increase in the risk of axial fractures but no increase in limb fractures. The pathophysiologic basis for the increased axial fractures should be determined. INTRODUCTION: Multiple myeloma is often preceded by monoclonal gammopathy of undetermined significance (MGUS). Fractures are common in myeloma as a result of lytic bone lesions, generalized bone loss, and elevated bone turnover from excessive cytokine production. Whether fractures are also increased in MGUS is unknown. MATERIALS AND METHODS: In a population-based retrospective cohort study, 488 Olmsted County, MN, residents with MGUS first diagnosed in 1960-1994 (52% men; mean age, 71.4 +/- 12.8 years) were followed for 3901 person-years; follow-up was censored at progression to myeloma. The relative risk of fractures was assessed by standardized incidence ratios (SIRs), and risk factors were evaluated in proportional hazards models. RESULTS AND CONCLUSIONS: Altogether, 200 patients experienced 385 fractures. Compared with expected rates in the community, statistically significant increases were seen for fractures at most axial sites, for example, vertebrae (SIR, 6.3; 95% CI, 5.2-7.5). There was a slight increase in hip (SIR, 1.6; 95% CI, 1.2-2.2) but not distal forearm fractures (SIR, 0.8; 95% CI, 0.4-1.5). The relative risk (SIR) of any axial fracture was 2.7 (95% CI, 2.3-3.1) compared with only 1.1 (95% CI, 0.9-1.4) for all limb fractures combined. In a multivariate analysis, the independent predictors of any subsequent fracture were age (hazard ratio [HR] per 10-year increase, 1.4; 95% CI, 1.2-1.6) and corticosteroid use (HR, 1.8; 95% CI, 1.2-2.6); greater weight at diagnosis (HR per 10 kg, 0.8; 95% CI, 0.8-0.9), and IgG monoclonal protein (HR, 0.7; 95% CI, 0.5-0.97) were protective. Baseline monoclonal protein level, a determinant of myeloma progression, did not predict fracture risk. Thus, the risk of axial, but not peripheral, fractures is increased among MGUS patients even before progression to myeloma. The pathophysiologic basis for this should be determined because elevated bone turnover, for example, might be treatable. PMID- 14753734 TI - Characterization of common genetic variants in cathepsin K and testing for association with bone mineral density in a large cohort of perimenopausal women from Scotland. AB - BMD values in approximately 3000 perimenopausal Scottish women were adjusted by regression to identify and account for nongenetic factors. Adjusted BMD values were not associated with simple tandem repeat (STR) markers or single nucleotide polymorphisms (SNPs) at the Cathepsin K (CTSK) locus. We present a thorough analysis of common CTSK polymorphisms and genetic relatedness among CTSK haplotypes. INTRODUCTION: CTSK is a cysteine protease of the papain family and is thought to play a critical role in osteoclast-mediated bone degradation. Rare, inactivating mutations in CTSK cause pychodysostosis, an autosomal recessive osteochondrodysplasia characterized by osteosclerosis and short stature. However, there have been no studies of common genetic variants in CTSK and their possible association with bone density in the general population. MATERIALS AND METHODS: To identify common single nucleotide polymorphisms (SNPs) and simple tandem repeat (STR) polymorphisms in and around CTSK, we screened all CTSK exons, intron A, all intron-exon boundaries, and the putative CTSK promoter region in 130 random whites using both high-performance liquid chromatography (HPLC) and DNA sequencing. CTSK markers were genotyped in approximately 3000 perimenopausal Scottish women whose hip and spine bone mineral density (BMD) had been measured by DXA. We performed linear regression analysis to identify and adjust for nongenetic predictors of BMD, and adjusted BMD values (regression residuals) were tested for association with individual CTSK markers and haplotypes by ANOVA and the composite haplotype method of Zaykin et al. RESULTS AND CONCLUSIONS: We discovered two intronic SNPs (8% and 9% frequency), but no common exonic SNPs (> 1% frequency), and found that three STRs at the immediate 5' end of the CTSK locus are highly polymorphic. The population frequencies of haplotypes defined by these five polymorphisms were estimated, and a cladogram was derived showing proximity of relationship and likely descent of the 30 most common CTSK haplotypes. Regression analyses revealed that approximately 39% of spine and 19% of hip rate of change in BMD was accounted for by nongenetic factors. For baseline BMD values in premenopausal women, nongenetic predictors explained 11% of the variance at the spine and 13% at the hip. Adjusted BMD values showed no statistically significant association with any of the individual CTSK polymorphisms or CTSK haplotypes. PMID- 14753735 TI - Genetic predisposition for adult lactose intolerance and relation to diet, bone density, and bone fractures. AB - Evidence that genetic disposition for adult lactose intolerance significantly affects calcium intake, bone density, and fractures in postmenopausal women is presented. PCR-based genotyping of lactase gene polymorphisms may complement diagnostic procedures to identify persons at risk for both lactose malabsorption and osteoporosis. INTRODUCTION: Lactase deficiency is a common autosomal recessive condition resulting in decreased intestinal lactose degradation. A 13910 T/C dimorphism (LCT) near the lactase phlorizin hydrolase gene, reported to be strongly associated with adult lactase nonpersistence, may have an impact on calcium supply, bone density, and osteoporotic fractures in the elderly. MATERIALS AND METHODS: We determined LCT genotypes TT, TC, and CC in 258 postmenopausal women using a polymerase chain reaction-based assay. Genotypes were related to milk intolerance, nutritional calcium intake, intestinal calcium absorption, bone mineral density (BMD), and nonvertebral fractures. RESULTS: Twenty-four percent of all women were found to have CC genotypes and genetic lactase deficiency. Age-adjusted BMD at the hip in CC genotypes and at the spine in CC and TC genotypes was reduced by -7% to -11% depending on the site measured (p = 0.04). LCT(T/C-13910) polymorphisms alone accounted for 2-4% of BMD in a multiple regression model. Bone fracture incidence was significantly associated with CC genotypes (p = 0.001). Milk calcium intake was significantly lower (-55%, p = 0.004) and aversion to milk consumption was significantly higher (+166%, p = 0.01) in women with the CC genotype, but there were no differences in overall dietary calcium intake or in intestinal calcium absorption test values. CONCLUSION: The LCT(T/C-13910) polymorphism is associated with subjective milk intolerance, reduced milk calcium intake, and reduced BMD at the hip and the lumbar spine and may predispose to bone fractures. Genetic testing for lactase deficiency may complement indirect methods in the detection of individuals at risk for both lactose malabsorption and osteoporosis. PMID- 14753736 TI - Trabecular bone microarchitecture is deteriorated in men with spinal cord injury. AB - Using magnetic resonance imaging, men with spinal cord injury (n = 10) were found to have fewer trabeculae that were spaced further apart in the knee than able bodied controls of similar age, height, and weight (n = 8). The deteriorated trabecular bone microarchitecture may contribute to the increased fracture incidence after injury. INTRODUCTION: Spinal cord injury results in a dramatic decline in areal bone mineral density (aBMD) and a marked increase in lower extremity fracture; however, its effect on trabecular bone microarchitecture is unknown. The purpose of this study was to determine if trabecular bone microarchitecture is deteriorated in the knee of men with long-term, complete spinal cord injury. MATERIALS AND METHODS: Apparent bone volume to total volume (appBV/TV), trabecular number, (appTb.N), trabecular thickness (appTb.Th), and trabecular separation (appTb.Sp), measures of trabecular bone microarchitecture, were assessed in the distal femur and proximal tibia of men with long-term (>2 years) complete spinal cord injury (SCI; n = 10) and able-bodied controls (CON; n = 8) using high-resolution magnetic resonance imaging. Proximal tibia and arm aBMD were determined using DXA. Independent t-tests were used to assess group differences in anthropometrics and bone parameters. Pearson correlation analysis was used to assess the relationships among trabecular bone microarchitecture, aBMD, and time since injury. RESULTS: There were no group differences in age, height, or weight; however, the distal femur and proximal tibia of SCI had 27% and 20% lower appBV/TV, 21% and 20% lower appTb.N, and 44% and 33% higher appTb.Sp, respectively (p < 0.05). The distal femur of SCI also had 8% lower appTb.Th (p < 0.05). Whereas arm aBMD was not different in the two groups, proximal tibia aBMD was 43% lower in SCI. In SCI and CON combined, aBMD was correlated with appBV/TV (r = 0.62), appTb.N (r = 0.78), and appTb.Sp (r = -0.82) in the proximal tibia (p < 0.05). Time since injury was more strongly correlated with appTb.N (r = -0.54) and appTb.Sp (r = 0.56) than aBMD (r = -0.36) in the distal tibia, although none of the relationships were statistically significant (p > 0.05). CONCLUSION: Men with complete spinal cord injury have markedly deteriorated trabecular bone microarchitecture in the knee, which may contribute to their increased fracture incidence. PMID- 14753737 TI - Umbilical venous IGF-1 concentration, neonatal bone mass, and body composition. AB - IGF-1 is a key growth factor during fetal life. Using DXA, we found that the concentration of IGF-1 in umbilical cord serum is strongly related to neonatal whole body bone mineral content, lean mass, and fat mass. However IGF-1 did not explain the relationships of maternal smoking, fat mass, and physical activity with neonatal bone mass. The study supports a direct role for circulating IGF-1 in growth of the fetal skeleton. INTRODUCTION: Evidence is accumulating that the risk of osteoporosis in later life may be determined in part by environmental influences during intrauterine and early postnatal life. We previously reported that maternal birthweight, smoking, fat stores, and physical activity during pregnancy predict neonatal bone mass. While the growth hormone/insulin-like growth factor 1 (GH/IGF-1) axis is an important determinant of postnatal skeletal growth, there are few data relating the concentration of growth factors in umbilical cord blood to bone mineral content (BMC) and other indices of body composition in the newborn infant. MATERIALS AND METHODS: We conducted a population-based study in a cohort of full-term, newborn infants whose mothers were characterized for lifestyle, body composition, and nutrition through their normal pregnancies. In a sample of 119 infants from the cohort, we related cord serum IGF-1 and insulin-like growth factor binding protein (IGFBP)-3 concentrations to neonatal body composition measured by DXA and evaluated the extent to which this cytokine mediates the previously reported effects of maternal diet and lifestyle on neonatal bone mass. RESULTS: There were strong positive associations between cord serum IGF-1 concentration and whole body BMC (r = 0.38, p < 0.001), whole body lean mass (r = 0.40, p < 0.001), and whole body fat mass (r = 0.50, p < 0.001) after adjusting for gestational age and sex. There was no association between cord serum IGF-1 and BMC adjusted for bone size. Neither cord serum IGF-1 nor IGFBP-3 explained the relationships that we previously reported between maternal influences and neonatal bone mass. CONCLUSIONS: Cord serum IGF-1 is more closely related to the size of the neonatal skeleton than to its degree of mineralization. Documented maternal determinants of neonatal bone mass seem to mediate their effects independently of variations in cord serum IGF-1 in healthy pregnancies. PMID- 14753738 TI - Earliest mineral and matrix changes in force-induced musculoskeletal disease as revealed by Raman microspectroscopic imaging. AB - Craniosynostosis, premature fusion of the skull bones at the sutures, is the second most common human birth defect in the skull. Raman microspectroscopy was used to examine the composition, relative amounts, and locations of the mineral and matrix produced in mouse skulls undergoing force-induced craniosynostosis. Raman imaging revealed decreased relative mineral content in skulls undergoing craniosynostosis compared with unloaded specimens. INTRODUCTION: Raman microspectroscopy, a nondestructive vibrational spectroscopic technique, was used to examine the composition, relative amounts, and locations of the mineral and matrix produced in mouse skulls undergoing force-induced craniosynostosis. Craniosynostosis, premature fusion of the skull bones at the sutures, is the second most common birth defect in the face and skull. The calvaria, or flat bones that comprise the top of the skull, are most often affected, and craniosynostosis is a feature of over 100 human syndromes and conditions. MATERIALS AND METHODS: Raman images of the suture, the tips immediately adjacent to the suture (osteogenic fronts), and mature parietal bones of loaded and unloaded calvaria were acquired. Images were acquired at 2.6 x 2.6 microm spatial resolution and ranged in a field of view from 180 x 210 microm to 180 x 325 microm. RESULTS AND CONCLUSIONS: This study found that osteogenic fronts subjected to uniaxial compression had decreased relative mineral content compared with unloaded osteogenic fronts, presumably because of new and incomplete mineral deposition. Increased matrix production in osteogenic fronts undergoing craniosynostosis was observed. Understanding how force affects the composition, relative amounts, and location of the mineral and matrix provides insight into musculoskeletal disease in general and craniosynostosis in particular. This is the first report in which Raman microspectroscopy was used to study musculoskeletal disease. These data show how Raman microspectroscopy can be used to study subtle changes that occur in disease. PMID- 14753739 TI - Estrogen receptor-beta inhibits skeletal growth and has the capacity to mediate growth plate fusion in female mice. AB - To determine the long-term role of ER beta in the regulation of longitudinal bone growth, appendicular and axial skeletal growth was followed and compared in female ER beta-/-, ER alpha-/-, and ER alpha-/- beta-/- mice. Our results show that ER beta inhibits appendicular and axial skeletal growth and has the capacity to induce fusion of the growth plates. INTRODUCTION: Estrogen affects skeletal growth and promotes growth plate fusion in humans. In rodents, the growth plates do not fuse after sexual maturation, but prolonged treatment with supraphysiological levels of estradiol has the capacity to fuse the growth plates. It should be emphasized that the estrogen receptor (ER) alpha-/- and the ER alpha-/- beta-/-, but not the ER beta-/-, mouse models have clearly increased serum levels of estradiol. MATERIALS AND METHODS: The skeletal growth was monitored by X-ray and dynamic histomorphometry, and the growth plates were analyzed by quantitative histology, calcein double labeling, bromodeoxyuridine (BrdU) incorporation, and TUNEL assay in 4- and 18-month-old female ER beta-/-, ER alpha-/-, and ER alpha-/- beta-/- mice. RESULTS: Young adult (4-month-old) ER beta-/- mice demonstrated an increased axial- and appendicular-skeletal growth, supporting the notion that ER beta inhibits skeletal growth in young adult female mice. Interestingly, the growth plates were consistently fused in the appendicular skeleton of 18-month-old female ER alpha-/- mice. This fusion of growth plates, caused by a prolonged exposure to supraphysiological levels of estradiol in female ER alpha-/- mice, must be mediated through ER beta because old ER alpah-/- beta-/- mice displayed unchanged, unfused growth plates. CONCLUSIONS: Our results confirm that ER beta is a physiological inhibitor of appendicular- and axial-skeletal growth in young adult female mice. Furthermore, we made the novel observation that ER beta, after prolonged supraphysiological estradiol exposure, has the capacity to mediate growth plate fusion in old female mice. PMID- 14753740 TI - Identification of promoter regions involved in cell- and developmental stage specific osteopontin expression in bone, kidney, placenta, and mammary gland: an analysis of transgenic mice. AB - Cell-specific expression of GFP under the control of different lengths of the osteopontin promoter in transgenic mice identified the positive and negative regulatory regions for respective cell types. The results provide new insights for physiological and pathological expression of the osteopontin gene. INTRODUCTION: Osteopontin (OPN) is a major non-collagenous bone matrix protein that is involved in normal and pathological calcification and is expressed in a tissue-specific manner. To investigate how such tissue-specific OPN gene expression is regulated in vivo, transgenic mice expressing the green fluorescent protein (GFP) reporter gene controlled by different lengths of the OPN promoter were generated. MATERIALS AND METHODS: Cell- and developmental stage-specific osteopontin expression in transgenic mice was examined by Northern blotting, immunoblotting, fluorescence examination, and in situ hybridization and compared with those of OPN. RESULTS AND CONCLUSIONS: The line bearing the -5505 to +14 region of the OPN promoter was shown by Northern blotting and immunoblotting to express GFP in the same cells that express endogenous OPN (osteoblasts, hypertrophic chondrocytes, renal and mammary gland epithelial cells, and granulated metrial gland [GMG] placental cells) at the same stage in development. Thus, the 5.5-kb -5505 to +14 promoter region is sufficient for proper tissue specific OPN expression. The lines carrying shorter segments of the OPN promoter showed different expression patterns. These patterns revealed a putative cis acting element in the -5269 to -5263 region that restricts OPN expression to hypertrophic chondrocytes and a mammary gland-specific expressing element and a GMG cell-specific enhancing element in the -5505 to -3156 region. Furthermore, the -3155 to -1576 region seems to contain positive renal epithelial cell- and GMG cell-specific expression motif(s) as well as a negative regulatory element that prevents OPN expression in fibroblasts. Moreover, the -1576 to -910 region seems to contain a positive osteoblast-specific-expressing element. Thus, the 5.5 kb OPN promoter contains multiple cis-acting elements encoding positive and negative cell-specific regulatory systems. PMID- 14753741 TI - Isolation of a human homolog of osteoclast inhibitory lectin that inhibits the formation and function of osteoclasts. AB - Osteoclast inhibitory lectin (OCIL) is a newly recognized inhibitor of osteoclast formation. We identified a human homolog of OCIL and its gene, determined its regulation in human osteoblast cell lines, and established that it can inhibit murine and human osteoclast formation and resorption. OCIL shows promise as a new antiresorptive. INTRODUCTION: Murine and rat osteoclast inhibitory lectins (mOCIL and rOCIL, respectively) are type II membrane C-type lectins expressed by osteoblasts and other extraskeletal tissues, with the extracellular domain of each, expressed as a recombinant protein, able to inhibit in vitro osteoclast formation. MATERIALS AND METHODS: We isolated the human homolog of OCIL (hOCIL) from a human fetal cDNA library that predicts a 191 amino acid type II membrane protein, with the 112 amino acid C-type lectin region in the extracellular domain having 53% identity with the C-type lectin sequences of rOCIL and mOCIL. The extracellular domain of hOCIL was expressed as a soluble recombinant protein in E. coli, and its biological effects were determined. RESULTS AND CONCLUSIONS: The hOCIL gene is 25 kb in length, comprised of five exons, and is a member of a superfamily of natural killer (NK) cell receptors encoded by the NK gene complex located on chromosome 12. Human OCIL mRNA expression is upregulated by interleukin (IL)-1alpha and prostaglandin E2 (PGE2) in a time-dependent manner in human osteogenic sarcoma MG63 cells, but not by dexamethasone or 1,25 dihydroxyvitamin D3. Soluble recombinant hOCIL had biological effects comparable with recombinant mOCIL on human and murine osteoclastogenesis. In addition to its capacity to limit osteoclast formation, OCIL was also able to inhibit bone resorption by mature, giant-cell tumor-derived osteoclasts. Thus, a human homolog of OCIL exists that is highly conserved with mOCIL in its primary amino acid sequence (C-lectin domain), genomic structure, and activity to inhibit osteoclastogenesis. PMID- 14753742 TI - Paracrine and autocrine signals promoting full chondrogenic differentiation of a mesoblastic cell line. AB - The pluripotent mesoblastic C1 cell line was used under serum-free culture conditions to investigate how paracrine and autocrine signals cooperate to drive chondrogenesis. Sequential addition of two systemic hormones, dexamethasone and triiodothyronine, permits full chondrogenic differentiation. The cell intrinsic activation of the BMP signaling pathway and Sox9 expression occurring on mesoblastic condensation is insufficient for recruitment of the progenitors. Dexamethasone-dependent Sox9 upregulation is essential for chondrogenesis. INTRODUCTION: Differentiation of lineage stem cells relies on cell autonomous regulations modulated by external signals. We used the pluripotent mesoblastic C1 cell line under serum-free culture conditions to investigate how paracrine and autocrine signals cooperate to induce differentiation of a precursor clone along the chondrogenic lineage. MATERIALS AND METHODS: C1 cells, cultured as aggregates, were induced toward chondrogenesis by addition of 10(-7) M dexamethasone in serum-free medium. After 30 days, dexamethasone was replaced by 10 nM triiodothyronine to promote final hypertrophic conversion. Mature and hypertrophic phenotypes were characterized by immunocytochemistry using specific antibodies against types II and X collagens, respectively. Type II collagen, bone morphogenetic proteins (BMPs), BMP receptors, Smads, and Sox9 expression were monitored by reverse transcriptase-polymerase chain reaction (RT-PCR), Northern blot, and/or Western blot analysis. RESULTS AND CONCLUSIONS: Once C1 cells have formed nodules, sequential addition of two systemic hormones is sufficient to promote full chondrogenic differentiation. In response to dexamethasone, nearly 100% of the C1 precursors engage in chondrogenesis and convert within 30 days into mature chondrocytes, which triggers a typical cartilage matrix. On day 25, a switch in type II procollagen mRNA splicing acted as a limiting step in the acquisition of the mature chondrocyte phenotype. On day 30, substitution of dexamethasone with triiodothyronine triggers the final differentiation into hypertrophic chondrocytes within a further 15 days. The chondrogenic process is supported by intrinsic expression of Sox9 and BMP family genes. Similarly to the in vivo situation, activation of Sox9 expression and the BMP signaling pathway occurred on mesoblastic condensation. After induction, BMP-activated Smad nuclear translocation persisted throughout the process until the onset of hypertrophy. After dexamethasone addition, Sox9 expression was upregulated. Dexamethasone withdrawal reversed the increase in Sox9 expression and stopped differentiation. Thus, Sox9 seems to be a downstream mediator of dexamethasone action. PMID- 14753743 TI - Adenovirus mediated BMP-13 gene transfer induces chondrogenic differentiation of murine mesenchymal progenitor cells. AB - Chondrogenic/osteogenic differentiation of a mesenchymal progenitor stimulated by BMP-13 (CDMP-2) was studied. C3H10T1/2 cells were transduced by an adenoviral construct containing BMP-13 or BMP-2. BMP-13 supported chondrogenesis but not terminal differentiation, whereas BMP-2 stimulated endochondral ossification. The studies show that BMP-13 may fail to support terminal chondrocyte differentiation. INTRODUCTION: Bone morphogenetic protein (BMP)-13 is a member of the transforming growth factor beta (TGF-beta) superfamily of growth factors. Although the biological functions of BMP-13 remain poorly understood, continued postnatal expression of BMP-13 in articular cartilage suggests that this protein may function in an autocrine/paracrine fashion to regulate growth and maintenance of articular cartilage. The purpose of this study was to elucidate the role of BMP-13 in chondrogenic differentiation. MATERIALS AND METHODS: Replication deficient adenoviruses carrying human BMP-13 (Adv-hBMP13), bacterial beta galactosidase (Adv-beta gal), and human BMP-2 (Adv-hBMP2) were constructed. Murine mesenchymal progenitor cells (C3H10T1/2) were transduced with these vectors, and differentiation to the chondrogenic lineage was assessed by reverse transcriptase-polymerase chain reaction (RT-PCR), biochemical, and histological analyses. RESULTS AND CONCLUSIONS: Our findings revealed that hBMP-13 transduced cells differentiated into round cells that stained with Alcian blue. Analysis of gene expression in hBMP-13-transduced cells demonstrated presence of cartilage specific markers, absence of hypertrophic chondrocyte specific markers, and upregulation of proteoglycan biosynthesis. In particular, hBMP-13-transduced cells had significantly less and delayed expression of alkaline phosphatase activity and calcium mineral accumulation than hBMP-2-transduced cells. Except for BMPR-IB/ALK-6, expression of BMP receptors was identified constitutively in C3H10T1/2 cells and was not affected by the presence of either of the BMPs. In summary, hBMP-13, while stimulating chondrogenesis, failed to support differentiation to hypertrophic chondrocytes and endochondral ossification similar to hBMP-2. Thus, this may prove to be a useful strategy for cell-based regeneration of articular cartilage. PMID- 14753744 TI - Expression and activity of the CDK inhibitor p57Kip2 in chondrocytes undergoing hypertrophic differentiation. AB - Growth plates of p57-null mice exhibit several abnormalities, including loss of collagen type X (CollX) expression. The phenotypic consequences of p57 expression were assessed in an in vitro model of hypertrophic differentiation. Adenoviral p57 expression was not sufficient for CollX expression but did augment induction of CollX by BMP-2. INTRODUCTION: During hypertrophic differentiation, chondrocytes pass from an actively proliferative state to a postmitotic, hypertrophic phenotype. The induction of growth arrest is a central feature of this phenotypic transition. Mice lacking the cyclin dependent-kinase inhibitor p57Kip2 exhibit several developmental abnormalities including chondrodysplasia. Although growth plate chondrocytes in p57-null mice undergo growth arrest, they do not express collagen type X, a specific marker of the hypertrophic phenotype. This study was carried out to investigate the link between p57 expression and the induction of collagen type X in chondrocytes and to determine whether p57 overexpression is sufficient for the induction of hypertrophic differentiation. MATERIALS AND METHODS: Neonatal rat epiphyseal or growth plate chondrocytes were maintained in an aggregate culture model, in defined, serum-free medium. Protein and mRNA levels were monitored by Western and Northern blot analyses, respectively. Proliferative activity was assessed by fluorescent measurement of total DNA and by 3H-thymidine incorporation rates. An adenoviral vector was used to assess the phenotypic consequences of p57 expression. RESULTS AND CONCLUSIONS: During in vitro hypertrophic differentiation, levels of p57 mRNA and protein were constant despite changes in chondrocyte proliferative activity and the induction of hypertrophic-specific genes in response to bone morphogenetic protein (BMP)-2. Adenoviral p57 overexpression induced growth arrest in prehypertrophic epiphyseal chondrocytes in a dose-dependent manner but was not sufficient for the induction of collagen type X, either alone or when coexpressed with the related CDKI p21Cip1. Similar results were obtained with more mature tibial growth plate chondrocytes. p57 overexpression did augment collagen type X induction by BMP-2. These data indicate that p57-mediated growth arrest is not sufficient for expression of the hypertrophic phenotype, but rather it occurs in parallel with other aspects of the differentiation pathway. Our findings also suggest a contributing role for p57 in the regulation of collagen type X expression in differentiating chondrocytes. PMID- 14753745 TI - The effects of 1alpha,25-dihydroxyvitamin D3 on the expression of DNA replication genes. AB - To identify key genes in the antiproliferative action of 1,25(OH)2D3, MC3T3-E1 mouse osteoblasts were subjected to cDNA microarray analyses. Eleven E2F-driven DNA replication genes were downregulated by 1,25(OH)2D3. These results were confirmed by quantitative RT-PCR in different cell types, showing the general nature of this action of 1,25(OH)2D3. INTRODUCTION: 1Alpha,25-dihydroxyvitamin D3 [1,25(OH)2D3] has a potent antiproliferative action characterized by a blocked transition from the G1- to the S-phase of the cell cycle. This study aims to identify genes whose expression is markedly altered after 1,25(OH)2D3 treatment in parallel with or preceding the observed G1-arrest. MATERIALS AND METHODS: The cDNA microarray technique was used, and the expression of approximately 4600 genes in MC3T3-E1 mouse osteoblasts was studied 6 and 12 h after treatment with 10(-8) M 1,25(OH)2D3. Quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) analyses were performed on MC3T3-E1 cells and on wildtype and vitamin D receptor (VDR) knockout primary murine epidermal keratinocytes (VDRwt MEKs, VDR-/- MEKs) and murine mammary tumor cells (GR) to confirm the microarray data. RESULTS AND CONCLUSIONS: After 12 h of treatment, in parallel with the 1,25(OH)2D3-induced G1 arrest, a particular set of DNA replication genes including a cell division cycle 6 homolog, a DNA polymerase alpha subunit, proliferating cell nuclear antigen, two DNA polymerase delta subunits, and flap structure specific endonuclease 1, was downregulated at least 2-fold. These genes are known targets of the E2F family of transcription factors, which are probably the central mediators of this action of 1,25(OH)2D3. Indeed, as shown by transfection assays with an E2F reporter construct, 12- and 24-h treatment of MC3T3-E1 cells with 1,25(OH)2D3 reduced E2F activity by 49% and 73%, respectively. Quantitative RT-PCR analyses confirmed the downregulation of these DNA replication genes by 1,25(OH)2D3 in MC3T3-E1, GR, and VDRwt MEKs cells, but not in VDR-/- MEKs cells, showing that this 1,25(OH)2D3-driven antiproliferative action is of a general nature and depends on a functional VDR. PMID- 14753746 TI - The nitrogen-containing bisphosphonate, zoledronic acid, influences RANKL expression in human osteoblast-like cells by activating TNF-alpha converting enzyme (TACE). AB - Bisphosphonates are used to prevent osteoclast-mediated bone loss. Zoledronic acid inhibits osteoclast maturation indirectly by increasing OPG protein secretion and decreasing transmembrane RANKL expression in human osteoblasts. The decreased transmembrane RANKL expression seems to be related to the upregulation of the RANKL sheddase, TACE. INTRODUCTION: Bisphosphonates (BPs) exhibit high affinity for hydroxyapatite mineral in bone and are used extensively to treat malignancy-associated bone disease and postmenopausal bone loss by inhibiting osteoclast (OC)-mediated bone resorption. MATERIALS AND METHODS: We examined the effect of the most potent nitrogen-containing BP available, zoledronic acid (ZOL), on the expression of RANKL and osteoprotegerin (OPG), critical factors in the regulation of OC formation and activation, in primary osteoblast (OB)-like cells derived from human bone, using flow cytometry, ELISA, semiquantitative reverse transcriptase-polymerase chain reaction (RT-PCR), in situ immunofluorescence staining, and Western blotting. RESULTS: Our studies show that ZOL, while not significantly affecting RANKL or OPG gene expression, markedly increased OPG protein secretion and reduced transmembrane RANKL protein expression in OB-like cells. The reduction in transmembrane RANKL expression was preceded by a marked increase in the expression of the metalloprotease disintegrin, TNF-alpha converting enzyme (TACE). In addition, the decreased transmembrane expression of RANKL could be partially reversed by a TACE inhibitor, TAPI-2. CONCLUSIONS: Our studies indicate that ZOL, in addition to its direct effects on mature OCs, may inhibit the recruitment and differentiation of OCs by cleavage of transmembrane RANKL in OB-like cells by upregulating the sheddase, TACE. PMID- 14753748 TI - Adult-onset idiopathic progressive acro-osteolysis with proximal symphalangism. AB - We experienced a 57-year-old female with adult-onset non-congenital idiopathic acro-osteolysis combined with proximal symphalangism. At the age of 36, she developed severe pain and swelling of the toe base of both feet and underwent Clayton surgery. However, the size of her toes diminished progressively over the 5-year period after surgery. At the age of 41, she suffered pain and swelling of the proximal interphalangeal (PIP) joints of fingers of both hands. These PIP joints became rigid and inflexible. Subsequently, she noticed shortening of the little finger of both hands, followed later by shortening of the index, middle, and ring fingers. At the age of 57, the thumbs began to shorten. Laboratory and endocrinological examinations were not abnormal. Finally, we diagnosed her with acro-osteolysis combined with proximal symphalangism by radiological examination. In this case, previously unreported mutations of the Noggin gene were identified. This is the first case report of adult-onset, non-congenital idiopathic acro osteolysis combined with proximal symphalangism. PMID- 14753747 TI - Selective blockade of voltage-gated potassium channels reduces inflammatory bone resorption in experimental periodontal disease. AB - The effects of the potassium channel (Kv1.3) blocker kaliotoxin on T-cell mediated periodontal bone resorption were examined in rats. Systemic administration of kaliotoxin abrogated the bone resorption in conjunction with decreased RANKL mRNA expression by T-cells in gingival tissue. This study suggests a plausible therapeutic approach for inflammatory bone resorption by targeting Kv1.3. INTRODUCTION: Kv1.3 is a critical potassium channel to counterbalance calcium influx at T-cell receptor activation. It is not known if Kv1.3 also regulates RANKL expression by antigen-activated T-cells, and consequently affects in vivo bone resorption mediated by activated T-cells. MATERIALS AND METHODS: Actinobacillus actinomycetemcomitans 29-kDa outer membrane protein-specific Th1-clone cells were used to evaluate the expression of Kv1.3 (using reverse transcriptase-polymerase chain reaction [RT-PCR] and Western blot analyses) and the effects of the potassium channel blocker kaliotoxin (0-100 nM) on T-cell activation parameters ([3H]thymidine incorporation assays and ELISA) and expression of RANKL and osteoprotegerin (OPG; flow cytometry, Western blot, and RT-PCR analyses). A rat periodontal disease model based on the adoptive transfer of activated 29-kDa outer membrane protein-specific Th1 clone cells was used to analyze the effects of kaliotoxin in T-cell-mediated alveolar bone resorption and RANKL and OPG mRNA expression by gingival T-cells. Stimulated 29 kDa outer membrane protein-specific Th1 clone cells were transferred intravenously on day 0 to all animals used in the study (n = 7 animals per group). Ten micrograms of kaliotoxin were injected subcutaneously twice per day on days 0, 1, 2, and 3, after adoptive transfer of the T-cells. The control group of rats was injected with saline as placebo on the same days as injections for the kaliotoxin-treated group. The MOCP-5 osteoclast precursor cell line was used in co-culture studies with fixed 29-kDa outer membrane protein-specific Th1-clone cells to measure T-cell-derived RANKL-mediated effects on osteoclastogenesis and resorption pit formation assays in vitro. Statistical significance was evaluated by Student's t-test. RESULTS: Kaliotoxin decreased T-cell activation parameters of 29-kDa outer membrane protein-specific Th1 clone cells in vitro and in vivo. Most importantly, kaliotoxin administration resulted in an 84% decrease of the bone resorption induced in the saline-treated control group. T-cells recovered from the gingival tissue of kaliotoxin-treated rats displayed lower ratios of RANKL and OPG mRNA expression than those recovered from the control group. The ratio of RANKL and osteoprotegerin protein expression and induction of RANKL dependent osteoclastogenesis by the activated T-cells were also markedly decreased after kaliotoxin treatments in vitro. CONCLUSION: The use of kaliotoxin or other means to block Kv1.3 may constitute a potential intervention therapy to prevent alveolar bone loss in periodontal disease. PMID- 14753749 TI - Seasonal variation of bone turnover? PMID- 14753750 TI - Biomarkers in breath condensate: a promising new non-invasive technique in free radical research. AB - Oxidative stress is associated with a range of inflammatory lung diseases including asthma, adult respiratory distress syndrome, idiopathic pulmonary fibrosis, pneumonia, lung transplantation, chronic obstructive pulmonary disease, cystic fibrosis, bronchiectasis and lung cancer. Increased concentrations of reactive oxygen species (ROS) in the airways of such patients are reflected by elevated concentrations of oxidative stress markers in the breath, airways, lung tissue and blood. Traditionally, the measurement of these biomarkers has involved invasive procedures to procure the samples, or examine the compartments. As a consequence, there is a need for less invasive approaches to measure oxidative stress. Analysis of breath hydrocarbons has partly fulfilled this need, however only gas phase volatile constituents can be assessed by this approach. The collection of exhaled breath condensate (EBC) is a simple, non-invasive approach, which comprehensively samples the lower respiratory tract. It is currently used as a research and diagnostic tool in the free radical field, yielding information on redox disturbance and the degree and type of inflammation in the lung. With further technical developments, such an approach may ultimately have a role in the clinic, in helping to diagnose specific lung diseases. EBC can be exploited to assess a spectrum of potential biomarkers, thus generating a "finger print" characteristic of the disease. By assessing the nature of oxidative stress in this manner, the most appropriate therapy can be selected and the response to treatment monitored. PMID- 14753751 TI - Aspirin prevents apoptosis and NF-kappaB activation induced by H2O2 in hela cells. AB - The classical pathway of nuclear factor-kappa B (NF-kappaB) activation by several inducers mainly involves the phosphorylation of IkappaBalpha by a signalsome complex composed of IkappaBalpha kinases (IKKalpha and IKKbeta). However, in some cell types hydrogen peroxide (H2O2) has been shown to activate an alternative pathway that does not involve the classical signalsome activation process. In this study, we demonstrate that H2O2 induced NF-kappaB activation in HeLa cells through phosphorylation and degradation of IkappaB proteins as shown by immunblot analysis. Our studies reveal that a commonly used non-steroid anti-inflammatory drug, acetylsalicylic acid (aspirin) prevents H2O2-induced NF-kappaB activation in a dose-dependent manner through inhibition of phosphorylation and degradation of IkappaBalpha and IkappaBbeta. Differential staining and DNA fragmentation analysis also show that aspirin preloading of HeLa cells also prevents H2O2 induced apoptosis in a dose-dependent manner with maximum efficiency at 10 mM concentration. Additionally, aspirin effectively prevents caspase-3 and caspase-9 (cysteinyl aspartate-specific proteases) activation by H2O2. These results suggest that NF-kappaB activation is involved in H2O2-induced apoptosis and aspirin may inhibit both processes simultaneously. PMID- 14753752 TI - Cholesterol, linoleic acid or/and tyrosine yield different spectra of products when oxidized alone or in a mixture: studies in various oxidative systems. AB - Identification of reliable biomarkers for oxidative stress for the prediction of the early development of pathological conditions is essential. The detection of biomarkers for oxidative stress such as degradation products of polyunsaturated fatty acid (PUFA), oxysterols, and oxidized proteins, as indicators of oxidative stress are in use, but suffers from insufficient specificity, accuracy and reliability. The overall aim of the present study was to develop new markers which will not only provide information about the presence and level of oxidative stress in biological systems but also on the type of reactive oxygen species (ROS) involved and their metabolic consequences. In the first stage of the study, we compared the level and type of oxidized products formed when different ROS were applied onto three major biomolecules, i.e. cholesterol, linoleic acid (LH) and tyrosine, representing sterols, PUFA and protein, when each compounds was exposed alone or in a mixture to the ROS [copper ions, 2,2-azobis(2 amidinopropane) dihydrochloride (AAPH) and hypochlorous acid (HOCl)]. It was found that different types of oxidants resulted in the formation of different types of oxidation products. Furthermore, oxidation pattern differs when the substrates (cholesterol, PUFA or amino acid) were present alone or in a mixture. As biological systems such as lipoproteins and cell membranes are composed of the above studied molecules, the need for simultaneous detection of the major oxidized products is requires for better characterization of the oxidative stress outcome. PMID- 14753753 TI - Acetaminophen toxicity in mice lacking NADPH oxidase activity: role of peroxynitrite formation and mitochondrial oxidant stress. AB - Previous data have indicated that activated macrophages may play a role in the mediation of acetaminophen toxicity. In the present study, we examined the significance of superoxide produced by macrophages by comparing the toxicity of acetaminophen in wild-type mice to mice deficient in gp91phox, a critical subunit of NADPH oxidase that is the primary source of phagocytic superoxide. Both groups of mice were dosed with 300 mg/kg of acetaminophen or saline and sacrificed at 1, 2, 4 or 24 h. Glutathione in total liver and in mitochondria was depleted by approximately 90% at 1 h in wild-type and knock out mice. No significant differences in toxicity (serum transaminase levels or histopathology) were observed between wild-type and mice deficient in gp91phox. Mitochondrial glutathione disulfide, as a percent of total glutathione, was determined as a measure of oxidant stress produced by increased superoxide, leading to hydrogen peroxide and/or peroxynitrite. The percent mitochondrial glutathione disulfide increased to approximately 60% at 1 h and 70% at 2 h in both groups of mice. Immunohistochemical staining for nitrotyrosine was present in vascular endothelial cells at 1 h in both groups of mice. Acetaminophen protein adducts were present in hepatocytes at 1 h in both wild-type and knock out animals. These data indicate that superoxide from activated macrophages is not critical to the development of acetaminophen toxicity and provide further support for the role of mitochondrial oxidant stress in acetaminophen toxicity. PMID- 14753754 TI - Benzo[a]pyrene enhances lipid peroxidation induced DNA damage in aorta of apolipoprotein E knockout mice. AB - The genotoxic compound benzo[a]pyrene (B[a]P) enhances atherosclerotic plaque progression, possibly by inducing oxidative stress and subsequent lipid peroxidation (LPO). Since LPO plays a key role in atherosclerosis, stable LPO derived DNA modifications such as 1,N6-ethenodeoxy-adenosine (epsilondA) and 3,N4 ethenodeoxy-cytidine (epsilondC) may be useful biomarkers for in vivo oxidative stress. In this study, benzo[a]pyrene-diol-epoxide (BPDE)-DNA, epsilondA and epsilondC were determined by 32P-postlabelling in apolipoprotein E knockout (ApoE KO) mice treated with 5mg/kg B[a]P by gavage. After 4 days, BPDE-DNA adduct levels were higher in aorta (10.8 +/- 1.4 adducts/10(8) nucleotides) than in lung (3.3 +/- 0.7, P < 0.05), which is a known target organ for B[a]P. Levels of epsilondA were higher in aorta of B[a]P-exposed animals than in unexposed controls (8.1 +/- 4.4 vs 3.4 +/- 2.1 adducts per 10(8) parent nucleotides, P < 0.05). On the other hand, epsilondC levels were not affected by B[a]P exposure. Serum low density lipoprotein (LDL) levels were lower in B[a]P-exposed mice than in controls (9.3 +/- 3.7 and 13.3 +/- 4.0mmol/l, respectively), whereas high density lipoprotein (HDL) levels were higher (1.4 +/- 1.6 and 0.4 +/- 0.3mmol/l, respectively). Consequently, a three-fold difference in the LDL/HDL ratio was observed (P = 0.001). epsilondA levels were positively related with plasma HDL concentrations (R = 0.68, P = 0.02), suggesting that the HDL mediated protection of the vessel wall against reactive lipid peroxides was reduced in B[a]P-exposed apoE-KO mice. Our observations show that direct as well as lipid peroxidation induced DNA damage is formed by B[a]P in aorta of apoE-KO mice, which may be involved in atherosclerotic plaque progression. This study further indicates that etheno-DNA adducts are useful biomarkers for in vivo oxidative stress in atherosclerosis. PMID- 14753755 TI - Oxidative stress-related alteration of the copy number of mitochondrial DNA in human leukocytes. AB - The role of oxidative stress in the regulation of the copy number of mitochondrial DNA (mtDNA) in human leukocytes is unclear. In this study, we investigated the redox factors in plasma that may contribute to the alteration of mtDNA copy number in human leukocytes. A total of 156 healthy subjects of 25-80 years of age who exhibited no significant difference in the distribution of subpopulations of leukocytes in blood were recruited. Small-molecular-weight antioxidants and thiobarbituric acid reactive substances (TBARS) in plasma and 8 hydroxy-2'-deoxyguanosine (8-OHdG) and 4,977bp deletion of mtDNA in leukocytes were determined. The mtDNA copy number in leukocytes was determined by real-time PCR. The results showed that the copy number of mtDNA in leukocytes was changed with age in a biphasic manner that fits in a positively quadratic regression model (P = 0.001). Retinol (P = 0.005), non-protein thiols (P = 0.001) and ferritin (P = 0.004) in plasma and total glutathione in erythrocytes (P = 0.046) were the significant redox factors that correlated with the mtDNA copy number in leukocytes in a positive manner. By contrast, alpha-tocopherol levels in plasma (P = 0.001) and erythrocytes (P = 0.033) were negatively correlated with the mtDNA copy number in leukocytes. Three oxidative indices including the incidence of 4,977 bp deletion of mtDNA (P = 0.016) and 8-OHdG content in leukocytes (P = 0.003) and TBARS in plasma (P = 0.001) were all positively correlated with the copy number of mtDNA in leukocytes. Taken these findings together, we suggest that the copy number of mtDNA in leukocytes is affected by oxidative stress in blood circulation elicited by the alteration of plasma antioxidants/prooxidants and oxidative damage to DNA. PMID- 14753756 TI - Oxidative inactivation of paraoxonase1, an antioxidant protein and its effect on antioxidant action. AB - Paraoxonase1 (PON1), one of antioxidant proteins to protect low density lipoprotein (LDL) from the oxidation, is known to lose its activity in the oxidative environment. Here, we attempted to elucidate the possible mechanisms for the oxidative inactivation of PON1, and to examine the capability of hydroxyl radicals-inactivated PON1 to prevent against LDL oxidation. Of various oxidative systems, the ascorbate/Cu2+ system was the most potent in inactivating the purified PON1 (PON1) as well as HDL-bound PON1 (HDL-PON1). In contrast to a limited inactivation by Fe2+ (2.0 microM), the inclusion of Cu2+ (0.1-1.0 microM) remarkably enhanced the inactivation of PON1 in the presence of ascorbate (0.5mM). A similar result was also obtained with the inactivation of HDL-PON1. The inactivation of PON1 by ascorbate/Cu2+ was pevented by catalase, but not general hydroxyl radical scavengers, supporting inactivation. In addition, Cu2+ alone inactivated PON1, either soluble or HDL-bound, by different mechanisms, concentration-dependent. Separately, there was a reverse relationship between the inactivation of PON1 and its preventive action against LDL oxidation during Cu2+ induced oxidation of LDL. Noteworthy, ascorbate/Cu2+-inactivated PON1, which was charaterized by the partial loss of histidine residues, expressed a lower protection against Cu2+-induced LDL oxidation, compared to native PON1. Based on these results, it is proposed that metal-catalyzed oxidation may be a primary factor to cause the decrease of HDL-associated PON1 activity under oxidative stress, and radicals-induced inactivation of PON1 may lead to the decrease in its antioxidant action against LDL oxidation. PMID- 14753757 TI - Human red blood cells as a natural flavonoid reservoir. AB - Quercetin is rapidly and avidly taken up by human red blood cells (RBC) via a passive diffusion mechanism, driven by flavonoid binding to haemoglobin and resulting in an almost quantitative accumulation of the flavonoid. Heamoglobin free resealed ghosts accumulated quercetin exclusively in the membrane fraction. Cell-associated quercetin was biological active and could be quantitatively utilised to support the reduction of extracellular oxidants mediated by a transplasma-membrane oxido-reductase. Additional experimental evidence revealed that quercetin uptake declined in the presence of albumin and that, under these conditions, the amount of cell-associated quercetin is enhanced by increasing the RBC number. Quercetin release from flavonoid-preloaded RBC was observed only in the presence of albumin (or in human plasma) and this response was progressively inhibited upon incubation in solutions containing albumin previously exposed to increasing concentrations of quercetin and cleared of the unbound fraction of the flavonoid. Furthermore, exposure to quercetin pre-saturated albumin promoted accumulation of the flavonoid in fresh RBC and this response was a direct function of the extent of albumin saturation. These results, indicating a flow of quercetin from albumin to haemoglobin, and vice versa, are therefore consistent with the possibility that human RBC play a pivotal role in the distribution and bioavailability of circulating flavonoids. PMID- 14753758 TI - Dental education and the Bologna Process. PMID- 14753759 TI - Effectiveness of interventions to promote continuing professional development for dentists. AB - BACKGROUND: Continuing education is incumbent upon dentists as health professionals, but its promotion may be required, particularly in order to ensure regular professional updating. Continuing professional development may be delivered in a variety of ways, and new strategies and techniques must be evaluated for effectiveness. AIM: To evaluate the effectiveness of two interventions utilizing the philosophies and techniques of the discipline of Quality Improvement. METHOD: A self-assessment instrument (a manual) for quality dental practice was developed using the Delphi technique. A randomized, controlled trial of the interventions was conducted under field conditions for dental practice in Victoria, Australia. Dentists in Test Groups 1 and 2 completed the self-assessment manual, and received relevant references and their own scores for the manual in comparison with empirical standards. Dentists in Test Group 1 also attended a continuing education course on Quality Improvement. Dentists in Control Group 1 completed the manual only and received feedback of their scores. Dentists in Test Groups 1 and 2, and in Control Group 1 completed the manual again after 1 year as a post-intervention follow-up. Dentists in Control Group 2 completed the manual only at 1 year. RESULTS: The intervention involving self assessment, receipt of scores and references for the manual resulted in modest improvements in total scores for dentists after 1 year, although a response bias was apparent. CONCLUSION: An effective method of facilitating change in quality dental practice was identified. Assessment of strategies and techniques for professional development of dentists should include observation of patterns of participation. PMID- 14753760 TI - Comprehensive validation of competencies for dental vocational training and general professional training. AB - This paper outlines a study designed to validate competencies for dental vocational training (DVT) and general professional training (GPT) in order to ensure their accuracy and acceptability. A highly inclusive approach is described whereby all trainers in Scotland were invited to participate in the exercise. The 168 individuals recruited were drawn from all branches of the dental services and all regions in Scotland. Using online or paper questionnaires, quantitative and qualitative data were collected for each competency statement over 9 months, after which focus groups discussed and decided which changes should be made. A high response rate was observed and from the 160 competencies originally identified, almost half (47.5%) were redrafted as a direct result of the validation process. Sections of the competency document that required most attention are discussed, as are the nature of changes made to the competencies. As a result of this study, a fully validated competency document for DVT and GPT has been produced and will allow a high degree of standardization of training through the provision of essential consistent information to trainers and VDPs. PMID- 14753761 TI - A virtual reality dental simulator predicts performance in an operative dentistry manikin course. AB - This study was designed to test the ability of a virtual reality dental simulator to predict the performance of students in a traditional operative dentistry manikin course. Twenty-six dental students were pre-tested on the simulator, prior to the course. They were briefly instructed and asked to prepare 12 class I cavities which were automatically graded by the simulator. The instructors in the manikin course that followed were unaware of the students' performances in the simulator pre-test. The scores achieved by each student in the last six simulator cavities were compared to their final comprehensive grades in the manikin course. Class standing of the students in the simulator pre-test positively correlated with their achievements in the manikin course with a correlation coefficient of 0.49 (P = 0.012). Eighty-nine percent of the students in the lower third of the class in the pre-test remained in the low performing half of the class in the manikin course. These results indicate that testing students in a dental simulator, prior to a manikin course, may be an efficient way to allow early identification of those who are likely to perform poorly. This in turn could enable early allocation of personal tutors to these students in order to improve their chances of success. PMID- 14753762 TI - A 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. Two groups of dental students, with no experience in operative dentistry, were trained solely by either VR or conventional training in the preparation of conventional class 1 cavities. The subjects all used the same operative armamentarium and phantom heads, and were allocated the same duration of practice periods. At the completion of these training periods, both groups produced two class 1 cavities on the lower left first molar, which were subsequently coded and blindly scored for the traditional assessment criteria of outline form, retention form, smoothness, cavity depth and cavity margin angulation. An ordinal score of 0-3 or 0-4 was assigned for each assessment criterion: the higher the score, the worse the evaluation. After initial independent scoring, the two examiners discussed any notable differences until an agreed score was reached. Once the codes were broken, non-parametric analyses were performed on the data. Wilcoxon Tests for the semiquantitative scores indicated significant differences between the VR and conventional training groups for outline form, depth and smoothness but not for retention or cavity margin angulation at P < 0.05 level, with the VR group receiving the higher, i.e. worse, scores. Cavity margin angulation approached significance with a P-value of 0.0536. The results indicated that VR-based skills acquisition is unsuitable for use as the sole method of feedback and evaluation for novice students. PMID- 14753763 TI - A comparison of sensation-seeking between dental and biological science students. AB - AIM: The aim of this study was to compare sensation-seeking behaviour amongst dental students and age-, sex-matched students studying for a standard 3-year degree in biological sciences at the Manchester University. METHODS: A total of 268 sensation-seeking questionnaires were distributed to second- and third-year male and female dental and biological science students, studying for a degree at the University of Manchester. Each questionnaire was scored against Zuckerman's sensation-seeking scale. RESULTS: A total of 268 questionnaires were distributed, 175 were returned. The return rate was 65%. The primary findings were: Biological Science students were more sensation-seeking in the dimensions of disinhibition and experience-seeking than dental students (P < 0.05). There was a significant gender bias in sensation-seeking. Males were significantly more sensation-seeking than females in both courses (P < 0.05). Males in both courses had similar sensation-seeking tendencies (NS). Females in the biological sciences course were less inhibited than their dental counterparts (P < 0.05). CONCLUSIONS: There was a clear difference in sensation-seeking within and between groups of dental students and students studying biological sciences. PMID- 14753764 TI - Time and cohort changes in preventive practice among Mongolian dental students. AB - To evaluate time and cohort changes in preventive practice of dental students in Mongolia, a questionnaire was distributed to all clinical-year students in 2000 (n = 79) and in 2002 (n = 73) of the Dental School of the Mongolian National Medical University (MNMU). The questionnaire eliciting information on students' preventive care-related practice and knowledge, oral self-care (OSC) behaviour, and self-perceived competency, was administered to the students. Further, cross sectional (between cohorts) comparisons to study changes over time and longitudinal (within-cohort) comparisons to reveal changes resulting from professional dental education were carried out. Statistical evaluations included chi-square test and linear regression model. Preventive practice of the students in 2002 tended to be better than that of their counterparts in the same study year in 2000. Preventive practice among the fifth-year students improved significantly compared to that in their third year, except for counselling on nutrition. Survey-year, study-year, and knowledge of and self-reported competency in preventive care explained 38% of the variation in reported preventive practice. Besides the positive changes revealed, preventive practice among the students could still be improved. Regarding these tasks, professional education should provide meaningful learning experience on modern caries-preventive methods and dental community should support practice of preventive dentistry. A special programme could help to motivate the students to improve their own OSC. PMID- 14753765 TI - Determination of endocrine disrupting chemicals in environmental solid matrices by extraction with a non-ionic surfactant (Tween 80). AB - A readily applicable method based on extraction by aqueous non-ionic surfactant solutions (Tween 80) and RP-HPLC coupled to fluorescence detection, has been developed for the simultaneous determination of the phenolic endocrine disrupting chemicals (EDCs) nonylphenol (NP), nonylphenol monoethoxylate (NP1EO) and nonylphenol diethoxylate (NP2EO) and bisphenol A (BPA) in environmental solid matrices. Clean up of sample extracts was performed on Si-C18 solid phase extraction (SPE) cartridges. The overall Tween 80 extraction-SPE-RP-HPLC procedure was validated for accuracy and precision by analyzing sediment samples spiked with known amounts of EDCs. Recoveries for NP, NP1EO, NP2EO and BPA and limits of detection are in agreement with conventional extraction methods. The developed methodology was successfully applied to the analysis of target compounds in Italian river sediments, river suspended matter and benthonic macroinvertebrate organisms (oligochaetes Lumbriculus variegatus). Results confirmed that this relatively simple procedure performed satisfactorily in the determination of phenolic EDCs in environmental solid matrices of different complexity and that it can be a suitable alternative method to conventional systems even for routine analyses. PMID- 14753767 TI - Prediction of peak shape as a function of retention in reversed-phase liquid chromatography. AB - Optimisation of the resolution of multicomponent samples in HPLC is usually carried out by changing the elution conditions and considering the variation in retention of the analytes, to which a standard peak shape is assigned. However, the change in peak shape with the composition of the mobile phase can ruin the optimisation process, yielding unexpected overlaps in the experimental chromatograms for the predicted optimum, especially for complex mixtures. The possibility of modelling peak shape, in addition to peak position, is therefore attractive. A simple modified-Gaussian model with a parabolic variance, which is a function of conventional experimental parameters: retention time (tR), peak height (H0), standard deviation at the peak maximum (sigma0), and left (A) and right (B) halfwidths, is proposed. The model is a simplification of a previous equation proposed in our laboratory. Linear and parabolic relationships were found between the peak shape parameters (sigma0), A and B) and tR, with a mean relative error of 1-5% in most cases. This error was partially due to variations in peak position and shape among injections, which in some cases were above 2%. Correlations between (sigma0, A and B) and the retention time, which is easily modelled as a function of mobile phase composition, allowed a simple and reliable prediction of chromatographic peaks. A parameter that depends on the slopes of the linear relationships for A and B versus tR is also proposed to evaluate column efficiency. The modified-Gaussian model was used to describe the peaks of six diuretics of diverse acid-base behaviour and polarity, which were eluted with 15 mobile phases where the composition was varied between 30 and 50% (v/v) acetonitrile and the pH between 3 and 7. PMID- 14753766 TI - Comparison of gas chromatography-mass spectrometry and capillary electrophoresis in analysis of phenolic compounds extracted from solid matrices with pressurized hot water. AB - Self-constructed pressurized hot water extraction (PHWE) equipment was used in dynamic mode to extract spiked phenolic compounds (phenol, 3-methylphenol, 4 chloro-3-methylphenol and 3,4-dichlorophenol) from sea sand and soil. Phenols were analyzed by both gas chromatography-mass spectrometry (GC-MS) and capillary zone electrophoresis (CZE) to compare the techniques and to find out if CZE is a suitable tool for analysis of phenols extracted from environmental matrix. Good recoveries of phenols spiked in sea sand were achieved at all PHWE temperatures (50, 100, 200, 300 C). GC-MS studies showed that phenols were selectively extracted from soil at 50 C but various other compounds (e.g. polyaromatic hydrocarbons) were extracted along with the phenols at 300 degrees C. In the case of CZE, phenols extracted from the soil, at 300 C were separated with good resolution at pH 9.7, and co-extracted compounds did not interfere with the analysis. The analytical values obtained by GC-MS and CZE were generally of similar magnitude. PMID- 14753768 TI - Determination of trace metallic impurities in high-purity quartz by ion chromatography. AB - A method has been developed for the determination of relevant trace impurities (alkali, alkaline and transition metals) in high purity quartz by ion chromatography. In situ reagent (HF) purification and simultaneous sample dissolution was achieved in a multichannel vapour phase digestion assembly. Twenty-one samples can be digested at a time in this vapour phase system. Significant decrease in the process blank levels for all the analytes was observed. Drastic reduction (250 times) of NH4+ blank was achieved in the described vapour phase digestion, which enables the determination of trace concentration of sodium in high purity quartz. After volatilisation of the matrix and unreacted HF, the clear water leached solutions were injected into an ion chromatograph equipped with conductivity detector for the determination of alkali and alkaline earth metals. In the case of transition metals, the trace residues were leached with 10 mM HCl and after separation on a mixed bed analytical column (IonPac CS5) were detected by spectrophotometry after post column derivatisation using 4-(2-pyridylazo)resorcinol (PAR). The accuracy of the result was checked by their comparison with those obtained by independent methods like inductively coupled plasma (ICP) MS and ICP atomic emission spectrometry. The achievable detection limits are between 0.4 ng/g (Li) and 22 ng/g (Mn). The application of the method to the determination of the above trace metals in two high-purity grade quartz samples is demonstrated. PMID- 14753769 TI - Preparation of hydrophobic interaction chromatographic packings based on monodisperse poly(glycidylmethacrylate-co-ethylenedimethacrylate) beads and their application. AB - The monodisperse, poly(glycidylmethacrylate-co-ethylenedimethacrylate) beads with macroporous in the range of 8.0-12.0 microm were prepared by a single-step swelling and polymerization method. The seed particles prepared by dispersion polymerization exhibited good absorption of the monomer phase. The pore size distribution of the beads was evaluated by gel permeation chromatography and mercury instrusion method. Based on this media, a hydrophobic interaction chromatographic (HIC) stationary phase for HPLC was synthesized by a new chemically modified method. The prepared resin has advantages for biopolymer separation, high column efficiency, low column backpressure, high protein mass recovery and good resolution for proteins. The dynamic protein loading capacity of the synthesized HIC packings was 40.0 mg/ml. Six proteins were fast separated in less than 8.0 min using the synthesized HIC stationary phase. The HIC resin was firstly used for the purification and simultaneous renaturation of recombinant human interferon-gamma (rhIFN-gamma) in the extract solution containing 7.0 mol/l guanidine hydrochloride with only one step. The purity and specific bioactivity of the purified of rhIFN-gamma was found more than 95% and 1.3 x 10(8) IU/mg, respectively. PMID- 14753770 TI - Comparative study of thiophilic functionalised matrices for polyclonal F(ab')2 purification. AB - Thiophilic adsorbents have been developed using divinyl sulfone or epoxy activated Streamline quartz base matrix. Their capacity and selectivity for binding polyclonal F(ab')2 fragments generated by whole serum proteolysis was tested. Except for epoxy activated guanidine, all the adsorbents displayed high selectivity for F(ab')2 with dynamic binding capacities ranging from 3 to 10 mg/ml of adsorbent. Thiol immobilised ligands adsorbed more F(ab')2 and the recovery was equal to or more than that from amino immobilised ligands. All adsorbents showed good selectivity for IgG and the dynamic binding capacities were better than for F(ab')2. PMID- 14753771 TI - Effects of pH and the presence of micelles on the resolution of diuretics by reversed-phase liquid chromatography. AB - A comparative study on the performance of two RPLC modes on the separation of 18 diuretics with diverse acid-base behaviour (acetazolamide, althiazide, amiloride, bendroflumethiazide, benzthiazide, bumetanide, canrenoic acid, chlorothiazide, chlorthalidone, ethacrynic acid, furosemide, hydrochlorothiazide, piretanide, probenecid, spironolactone, triamterene, trichloromethiazide and xipamide) was carried out. A conventional octadecylsilane column and acidic acetonitrile-water mobile phases, in the absence and presence of micelles of the anionic surfactant sodium dodecyl sulphate (SDS), were used. The effects of pH and the modifiers acetonitrile and SDS on peak asymmetry, efficiency, selectivity, resolution and analysis time, were examined. The comparison of both RPLC modes (aqueous- and micellar-organics) was done using the same processing tools, applying several polynomial and mechanistic equations to describe the retention. The best separations were obtained by maximising the product of peak purities, considering a wide range of experimental conditions. The study illustrates that, despite the theoretical and practical complexity of the problem, the predicted optimal chromatograms can be reproduced experimentally with great accuracy. None of the examined RPLC modes was able to yield baseline separation of the 18 diuretics. However, their selectivity was complementary, being appropriate for different combinations of a smaller number of the assayed diuretics. PMID- 14753772 TI - Analysis of polyphenolic antioxidants in star fruit using liquid chromatography and mass spectrometry. AB - Our previous study indicated that star fruit (Averrhoa carambola L.) is a very good source of natural antioxidants. However, it was still not clear which compounds were responsible for its antioxidant properties. The purpose of this study is to separate and identify compounds that contribute to total antioxidant activity in star fruit using HPLC and mass spectrometry (MS). HPLC coupled with a diode array detector (DAD) was used to characterise antioxidant peak in the juice or residue extract through spiking with free radicals. By analysing the antioxidant capacity and chromatograms of fractions from solid phase extraction, main antioxidants were attributed to phenolic compounds. The peaks were identified as L-ascorbic acid, (-)epicatechin and gallic acid in gallotannin forms. Other antioxidant peaks were further investigated using HPLC-ESI-MS-MS. Identification was confirmed with electronspray ionisation (ESI) MS-MS spectra of pure standards and singly-linked proanthocyanidins from pycnogenol. The major antioxidants were initially attributed to singly-linked proanthocyanidins that existed as dimers, trimers, tetramers and pentamers of catechin or epicatechin. PMID- 14753773 TI - Separation and purification of puerarin using beta-cyclodextrin-coupled agarose gel media. AB - The isoflavonoid puerarin, a well-known traditional Chinese drug, has been purified in one step from an extract of Radix puerariae (root of the plant Pueralria lobata) by adsorption chromatography on an epichlorohydrin polymerized beta-cyclodextrin ligand coupled to brominated allyl-group substituted Sepharose HP. Acetic acid (10%) was used as the mobile phase and the optimum loading capacity was around 1.2 mg crude extract/ml packed gel. The purity of the collected puerarin was about 98% with a recovery of about 62%. PMID- 14753774 TI - Use of reversed-phase high-performance liquid chromatography-diode array detection for complete separation of 2,4,6-trinitrotoluene metabolites and EPA Method 8330 explosives: influence of temperature and an ion-pair reagent. AB - Explosives such as 2,4,6-trinitrotoluene (TNT), octahydro-1,3,5,7-tetranitro 1,3,5,7-tetrazocine (HMX), and hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) are widely distributed environmental contaminants. Complete chromatographic separation is necessary in order to accurately determine and quantify explosives and their degradation products in environmental samples and in (bio)transformation studies. The present study describes a RP-HPLC method with diode array detection using a LC-8 guard column, a Supelcosil LC-8 chromatographic column, and a gradient elution system. This gradient method is capable of baseline separating the most commonly observed explosives and TNT transformation metabolites including 2,4,6-triaminotoluene (TAT) in a single run. The TNT metabolites separated were 2-hydroxylamino-4,6-dinitrotoluene, 4 hydroxylamino-2,6-dinitrotoluene, 2,4-dihydroxylamino-6-nitrotoluene, 4,4',6,6' tetranitro-2,2'-azoxytoluene, 2,2',6,6'-tetranitro-4,4'-azoxytoluene, 4,4',6,6' tetranitro-2,2'-azotoluene, 2,2',6,6'-tetranitro-4,4'-azotoluene, 2-amino-4,6 dinitrotoluene, 4-amino-2, 6-dinitrotoluene, 2,6-diamino-4-nitrotoluene, 2,4 diamino-6-nitrotoluene, and TAT. The same gradient method at a different column temperature can also be used to baseline separate the explosives targeted in the Environmental Protection Agency (EPA) Method 8330 with approximately 22% reduction in total run time and 48% decrease in solvent consumption compared to previously published methods. Good separation was also obtained when all TNT metabolites and EPA Method 8330 compounds (a total of 23 compounds) were analyzed together; only 2,6-DANT and HMX co-eluted in this case. The influence of temperature (35-55 degrees C) and the use of an ion-pair reagent on the chromatographic resolution and retention were investigated. Temperature was identified as the key parameter for optimal baseline separation. Increased temperature resulted in shorter retention times and better peak resolution especially for the aminoaromatics investigated. The use of an ion-pair reagent (octanesulfonic acid) generally resulted in longer retention times for compounds containing amine functional groups, more baseline noise, and decreased peak resolution. PMID- 14753775 TI - Analysis of aspartyl peptide degradation products by high-performance liquid chromatography and high-performance liquid chromatography-mass spectrometry. AB - A reversed-phase HPLC method for the analysis of degradation products of the model aspartyl tripeptides Phe-Asp-GlyNH2 and Gly-Asp-PheNH2 after incubation at pH 2 and 10 was developed. Most of the compounds could be separated with a gradient of acetonitrile in water containing 0.1% trifluoroacetic acid. Resolution of the isomeric pairs L-Phe-alpha-L-Asp-GlyNH2/L-Phe-beta-L-Asp-GlyNH2 and L-Phe-alpha-D-Asp-GlyOH/L-Phe-beta-D-Asp-GlyOH was achieved with a gradient of acetonitrile in phosphate buffer, pH 5.0. Under acidic conditions the major degradation pathway was cleavage of the peptide backbone amide bonds yielding dipeptides and amino acids, C-terminal deamidation as well as formation of succidinimyl peptides. At alkaline pH both deamidation of the C-terminal amide as well as isomerization and concomitant enantiomerization of Asp were observed. The peaks were identified both by reference substances and by online electrospray mass spectrometry. The results were compared to a previous developed capillary electrophoresis method. Diastereomeric pairs ofpeptides that could not be separated by capillary electrophoresis were resolved by HPLC while the separation of corresponding pairs of alpha- and beta-Asp peptides was not always achieved by HPLC in contrast to capillary electrophoresis illustrating that both techniques can be complimentary in peptide analysis. PMID- 14753776 TI - Continuous chromatographic protein refolding. AB - Column-based protein refolding requires a continuous processing capability if reasonable quantities of protein are to be produced. A popular column-based method, size-exclusion chromatography (SEC) refolding, employs size-exclusion matrices to separate unfolded protein from denaturant, thus refolding the protein. In this work, we conduct a comparison of SEC refolding with refolding by batch dilution, using lysozyme as a model protein. Lysozyme refolding yield was found to be extremely sensitive to the chemical composition of the refolding buffer and particularly the concentration of dithiothreitol (DTT) introduced from the denatured protein mixture. SEC refolding was not adversely affected by DTT carry-over as small contaminants in the denatured solution are separated from protein during the refolding operation. We also find that, contrary to previous reports, size-exclusion refolding on batch columns leads to refolding yields slightly better than batch dilution refolding yields at low protein concentrations but this advantage disappears at higher protein concentrations. As batch-mode chromatography would be the limiting step in a column based refolding downstream process, the batch column refolding method was translated to a continuously operating chromatography system (preparative continuous annular chromatography, P-CAC). It was shown that the P-CAC elution profile is similar to that of a stationary column, making scale-up and translation to P-CAC relatively simple. Moreover, it was shown that high refolding yields (72%) at high protein concentration (>1 mg ml(-1)) could be obtained. PMID- 14753777 TI - Simultaneous quantification of vitamins A, D3 and E in fortified infant formulae by liquid chromatography-mass spectrometry. AB - A novel method for the simultaneous quantification of Vitamins A, D3 and E in fortified infant formulae has been developed using isocratic normal-phase liquid chromatography with positive atmospheric pressure chemical ionization mass spectrometry (LC-APCI-MS). Food products were saponified and the vitamins were extracted by solid-phase extraction (SPE) on a Chromabond XTR cartridge. Quantification of Vitamins D3 and E were performed with Vitamin D2 and 5,7 dimethyltocol (DMT) as internal standards (IS), respectively while no IS was used for Vitamin A. Detection of the vitamins was made in the selected ion monitoring (SIM) mode. MS calibration curves were linear between 0.15 and 12 mg/l for Vitamin A, 5-400 microg/l for Vitamin D3 and 0.25-20 mg/l for Vitamin E with regression coefficient r2 > 0.996 and the limits of detection were below 1.4 ng. The repeatability (CV) obtained on a reference dietetic infant formula was 2.3% for Vitamin A, 2.6% for Vitamin E and 5.9% for Vitamin D3. The between-day variations (CV) over 6 days were in the ranges of 2.4-6.9% for the three vitamins. The mean recoveries from a reference infant formula spiked with all three vitamins ranged from 96 to 105% with a relative standard error less than 9%. The applicability of the method was demonstrated by analyzing a set of infant formula and infant cereals; similar results were obtained with the LC-MS method and reference HPLC methods. PMID- 14753778 TI - Liquid chromatography with ultraviolet absorbance detection for the analysis of tetracycline residues in honey. AB - The separation of tetracyclines (TCs) using reversed-phase liquid chromatography (LC) is proposed. The use of an amide-based stationary phase prevents the interaction of tetracyclines with the residual silanol groups and thus avoids the appearance of tailed peaks. Detection was based on using an UV spectrophotometer and gradient elution with acetonitrile-oxalic acid as mobile phase permitted good separation of all the peaks. Specificity was demonstrated by the retention characteristics, UV spectra and peak purity index. Linearity, precision, recovery and sensitivity were satisfactory. The procedure was applied to the analysis of tetracycline residues (tetracycline, oxytetracycline (OTC), chlortetracycline (CTC), doxycycline (DC), minocycline (MINO) and methacycline (MTC)) in honey of different types. Extraction involved using a mild acidic solvent containing EDTA to release protein-bound or sugar-bound tetracyclines. For the clean-up step, solid phase extraction using phenyl cartridges was applied. Detection limits in the honey using the proposed procedure are between 15 and 30 ng g(-1), depending on the tetracycline. PMID- 14753779 TI - High-performance liquid chromatographic-nuclear magnetic resonance investigation of the isomerization of alachlor-ethanesulfonic acid. AB - The metabolism of the acetanilide herbicide alachlor in soils leads to the formation of alachlor-ethanesulfonic acid (alachlor-ESA) as one of the major transformation products of this compound. The unique structure of alachlor and its metabolites allows the formation of two diastereomers (s-trans and s-cis) due to the hindered rotation of the amide bond connected to a rigid aromatic ring. Although these stereoisomers do interconvert by rotation about the amide bond, the rate of interconversion is slow allowing separation of the isomers on the chromatographic time scale. Once separated, the unique nuclear magnetic resonance signals of each isomer can be used to monitor the rate of isomerization. This paper reports the on-line separation and detection of the rotational diastereomers using high-performance liquid chromatography-nuclear magnetic resonance (HPLC-NMR) to efficiently measure the isomerization rate of alachlor ESA. PMID- 14753780 TI - Comparison of high-speed counter-current chromatography and high-performance liquid chromatography on fingerprinting of Chinese traditional medicine. AB - Active constituents of Chinese traditional medicine (CTM) are influenced by soils, climates, and growth stages. Therefore, fingerprint is important in quality control of CTM. Our studies showed for the first time that high-speed counter-current chromatography (HSCCC) could be applied in the development of fingerprint of CTM. Twelve components were separated from extracts of three crude samples of Salvia miltiorrhiza Bunge, which were collected from different growth locations. All 12 components of each sample were isolated within 13 h coming with good correspondence by using HSCCC. Relative standard deviation (R.S.D.) values of the retention time in HSCCC separation was less than 3% satisfied the precision requirements of national standards in the fingerprint development. As a new approach, HSCCC was compared to the conventional approach-HPLC in our experiments. The fingerprint developed by HPLC comprised 11 peaks that collected within 45 min. R.S.D. values of retention times of the corresponding peaks in HPLC analysis were very small (maximum 0.66% and average 0.10%). In conclusion, HSCCC was a feasible method to be used in the development of fingerprint of CTM. PMID- 14753781 TI - Rapid simultaneous determination of o-phenylphenol, diphenyl, thiabendazole, imazalil and its major metabolite in citrus fruits by liquid chromatography-mass spectrometry using atmospheric pressure photoionization. AB - A simple and rapid simultaneous analytical method of four post-harvest fungicides, o-phenylphenol (OPP), diphenyl (DP), thiabendazole (TBZ), imazalil (IMZ) and its major metabolite R14821 (IMZ-M) in citrus fruits was developed. These compounds were extracted under basic conditions with diethyl ether. The organic layer was washed with water and evaporated at moderate pressure adding methanol. These compounds were determined by liquid chromatography-mass spectrometry (LC-MS) using atmospheric pressure photoionization (APPI). The recoveries of the five compounds added to citrus fruits at 1 microg/g ranged from 67 to 100%, with relative standard deviations (R.S.D.) ranging from 2 to 8%. The detection limits (S/N = 3) were 0.01 microg/g and 0.05 microg/g (DP). PMID- 14753782 TI - Identification of organic compounds in atmospheric aerosol particles by on-line supercritical fluid extraction-liquid chromatography-gas chromatography-mass spectrometry. AB - Atmospheric particles were collected with a high-volume sampling system at an urban site in Helsinki (Finland). The samples were analysed by on-line coupled supercritical fluid extraction-liquid chromatography-gas chromatography-mass spectrometry (SFE-LC-GC-MS). The aerosol sample was first extracted by SFE. The extract was then transferred to a liquid chromatograph where it was fractionated into four fractions according to polarity. Each fraction from the liquid chromatograph was transferred to a gas chromatograph by large-volume injection, where final separation was carried out. The first LC fraction (280 microl) contained nonpolar compounds, such as n-alkanes, hopanes and steranes. The second fraction (840 microl) included polycyclic aromatic hydrocarbons (PAHs) and alkyl PAHs, while the third and fourth fractions (840 microl each) contained more polar compounds, such as n-alkan-2-ones, n-alkanals, oxy-PAHs and quinones. PMID- 14753783 TI - Application of liquid-phase microextraction and gas chromatography-mass spectrometry for the determination of polychlorinated biphenyls in blood plasma. AB - This study investigated the feasibility of applying liquid-phase microextraction combined with gas chromatography-mass spectrometry (GC-MS) to determine polychlorinated biphenyls (PCBs) in blood plasma. An efficient and simple extraction technique has been developed for the enrichment of PCBs from human blood plasma samples using single-step liquid-phase microextraction (LPME) in conjunction with a hollow fibre membrane (HFM). An eight PCB congener mixture was spiked into 2.5 ml of blood plasma, and the solution was then adjusted to pH 10.5 with a salinity of 20% (w/v) prior to making the total volume to 5 ml with ultrapure water. The porous HFM, filled with 3 microl of organic solvent, was then immersed into the solution, which was continuously agitated at 700 rpm for 30 min. Extract (1 microl) containing the pre-concentrated analytes was then injected into a GC-MS without further pre-treatment. Using an optimised extraction procedure, a large enrichment factor of the analytes, i.e. up to 241 fold was achieved in 30 min. The procedure resulted in a relative standard deviation of < 11% (n = 6), and a linear calibration range from 2.5 to 150 microg/l (r > 0.999), and detection limits between 0.07 and 0.94 microg/l, respectively. To demonstrate the feasibility of the procedure, PCB concentrations were determined in actual blood samples collected from the local population in Singapore using the optimised LPME technique. PMID- 14753784 TI - Development of a solid-phase extraction method for simultaneous extraction of adipic acid, succinic acid and 1,4-butanediol formed during hydrolysis of poly(butylene adipate) and poly(butylene succinate). AB - A solid-phase extraction (SPE) method was developed for the simultaneous extraction of dicarboxylic acids and diols formed during hydrolysis of poly(butylene succinate), PBS, and poly(butylene adipate), PBA. Four commercial non-polar SPE columns, three silica based: C8, C18, C18 (EC), and one resin based: ENV+, were tested for the extraction of succinic acid, adipic acid and 1,4 butanediol, the expected final hydrolysis products of PBS and PBA. ENV+ resin was chosen as a solid-phase, because it displayed the best extraction efficiency for 1,4-butanediol and succinic acid. Linear range for the extracted analytes was 1 500 ng/microl for adipic acid and 2-500 ng/microl for 1,4-butanediol and succinic acid. Detection and quantification limits for the analytes were between 1-2 and 2 7 ng/microl, respectively, and relative standard deviations were between 3 and 7%. Good repeatability and low detection limits made the developed SPE method and subsequent gas chromatography-mass spectrometry (GC-MS) analysis a sensitive tool for identification and quantification of hydrolysis products at early stages of degradation. PMID- 14753785 TI - Simultaneous determination of endocrine disrupting phenolic compounds and steroids in water by solid-phase extraction-gas chromatography-mass spectrometry. AB - A solid-phase extraction (SPE)-gas chromatography (GC)-mass spectrometry (MS) analytical method for the simultaneous separation and determination of endocrine disrupting chemicals (EDCs) from water samples is described in detail. Important and contrasting EDCs including estrone, 17beta-estradiol, 17beta ethynylestradiol, 16beta-hydroxyestrone, 4-nonylphenol, bisphenol A and 4-tert octylphenol were selected as the target compounds. The SPE technique, followed by the derivatisation with bis (trimethylsilyl) trifluoroacetamide was used for the extraction recoveries of target compounds from water samples. A number of parameters that may affect the recovery of EDCs, such as the type of SPE cartridges, eluents, as well as water properties including pH value, and concentration of salts and humic substances were investigated. It is shown that the Oasis cartridges produced the best recoveries of target EDCs while ethyl acetate was efficient in eluting EDCs from SPE cartridges. The recovery of some EDCs was enhanced by the addition of salt, but reduced by the increase in pH value and humic acid concentration. The optimised method was further verified by performing spiking experiments in natural river water and seawater matrices, with good recovery and reproducibility for all the selected compounds. The established method was successfully applied to environmental water samples from East and West Sussex, UK, for the determination of the target EDCs. PMID- 14753786 TI - Evaluation of weak ion association between tetraalkylammonium ions and inorganic anions in aqueous solutions by capillary zone electrophoresis. AB - The evaluation of weak ion association between eleven (11) inorganic anions (charge -1 to -3) and five n-tetraalkylammonium ions, R4N+ (R: methyl, Me; ethyl, Et; propyl, Pr; butyl, Bu; pentyl, Am) in aqueous media at 25 degrees C was studied. The analysis of ion association equilibria was carried out under acidic condition (formate buffer, pH 3.5) at low separating potential (-10 kV) using a coated capillary with suppressed electroosmotic flow (micro = 4 x 10(-5) cm2 V( 1) s(-1)). Direct UV detection was done at anode (lambda = 220 nm). The combination of the aforementioned conditions ensured that ion association constants, Kass, between n-tetraalkylammonium ion and the small inorganic anions were reliably determined after a non-linear least squares (NLLS) treatment of the measured anion's mobility. Like their larger counterparts, small anions showed increased interaction with an increase in size of pairing ions. Moreover, for a specific cation, the interaction of small anions increased with an increase in size of the hydrated anions as reflected by the relationship between the Kass and the Stokes' radius. A favourable comparison exists between the results presented in this work and those previously documented from other analytical techniques like conductometry. Qualitatively, the mobility of the anions appeared to obey the Huckel's model more closely than the more elaborate Zwanzig and Hubbard Onsager models. PMID- 14753787 TI - Post-column fluorescence derivatization of proteins and peptides in capillary electrophoresis with a sheath flow reactor and 488 nm argon ion laser excitation. AB - We report the use of a sheath flow reactor for post-column fluorescence derivatization of proteins. The derivatization reaction employed naphthalene-2,3 dicarboxaldehyde (NDA) and beta-mercaptoethanol, which were added in the sheath buffer. The labeled proteins were detected by laser-induced fluorescence with an argon-ion laser beam at 488 nm. The performance of this detection scheme was evaluated by separation of some protein standards. A column efficiency of 450,000 plates/m was obtained without stacking. The limits of detection for those standard proteins were determined to be from 8 to 32 nM. Excellent linear relationship was obtained with correlation coefficient of 0.9998 for alpha lactalbumin concentration ranging from 3.91 x 10(-7) to 1.25 x 10(-5) M. Separation of protein standards at low pH was also demonstrated by reversing the electroosmotic flow (EOF) with addition of cetyltrimethylammonium bromide (CTAB) to the running buffer. Different separation selectivity was achieved, but the sensitivity is poorer than that at high pH. This post-column derivatization detection system was applied successfully to analyze the protein extract from HT29 human colon cancer cells as well as tryptic peptides. PMID- 14753788 TI - Microchip capillary electrophoresis with a boron-doped diamond electrode for rapid separation and detection of purines. AB - Microchip capillary electrophoresis (CE) coupled with a boron-doped diamond (BDD) electrode has been employed for the separation and detection of several purines and purine-containing compounds. The BDD end-channel amperometric detector offers favorable signal-to-noise (S/N) characteristics at the high detection potential (+ 1.3 V) essential for detecting purine-related compounds. Factors influencing the separation and detection processes were examined and optimized. Five purines (guanine, hypoxanthine, guanosine, xanthine, and uric acid) have been separated within 6 min at a separation voltage of 1000V using a borate/phosphate run buffer (pH 8.2). Linear calibration plots are observed for micromolar concentrations of the purine compounds. Good stability and reproducibility (R.S.D. < 5%) are obtained reflecting the minimal adsorption of purines at the BDD surface. Applicability for the detection of nucleosides, nucleotides, and oligonucleotides is illustrated. The new microchip protocol offers great promise for a wide range of bioanalytical applications involving assays of purines and purine-containing compounds. PMID- 14753789 TI - Isolation of high purity 1-[2',4'-dihydroxy-3',5'-di-(3"-methylbut-2"-enyl)-6' methoxy] phenylethanone from Acronychia pedunculata (L.) Miq. by high-speed counter-current chromatography. AB - Following an initial clean-up step on silica, high-speed counter-current chromatography (HSCCC) was used to purify an aryl ketone, 1-[2',4'-dihydroxy 3',5'-di-(3"-methylbut-2"-enyl)-6'-methoxy] phenylethanone from an extract of the stem bark of the shrub Acronychia pedunculata. The two-phase solvent system used was composed of n-heptane-ethyl acetate-methanol-water at an optimized volume ratio of 4:1:4:1 (v/v/v/v). Target compound (58.1 mg) with a purity of 98.9% was obtained after HSCCC of 183.5 mg sample with a purity of 35.7% recovered after the silica clean-up step. Identification of the target compound was performed by 1H NMR, 13C NMR, two-dimensional NMR and LC-electrospray ionization MS. PMID- 14753790 TI - Simultaneous separation of nine metal ions and ammonium with nonaqueous capillary electrophoresis. AB - A simple and fast method for simultaneous separation of nine metal cations Ni2+, Cu2+, Co2+, Zn2+ Cd2+, K+, Na+, Mg2+ and Ca2+, and NH4+ in methanol is reported. The optimization for separation these 10 cations was achieved by using 0.5% acetic acid and 10 mM imidazole as electrolyte. The effects of water and ionic strength in the sample are discussed. The sensitive detection of transition metal ions was accomplished at 191 nm. The optimized method demonstrated high efficiency and good reproducibility, and was applied successfully to the qualitative and quantitative determination of transition metal ions in water samples, chemical reagents, oral zinc gluconate solution and human plasma. PMID- 14753791 TI - Multidetector-row computed tomography. PMID- 14753792 TI - Rotator cuff ultrasonography: diagnostic capabilities. AB - Dynamic shoulder ultrasonography is a noninvasive, accurate method used to assess the rotator cuff tendons. It can be incorporated into the in-office physical examination of patients at risk for rotator cuff problems. The low cost, convenience, and lack of risk make dynamic ultrasonography an excellent imaging tool for evaluating the biceps, subscapularis, infraspinatus, teres minor, and supraspinatus tendons. In-office ultrasound also provides an opportunity for patient education and explanation of management options. PMID- 14753793 TI - Periprosthetic femoral fractures above total knee replacements. AB - Periprosthetic femoral fractures above total knee replacements can be managed by a variety of methods, including casting, open reduction and internal fixation, external fixation, or revision arthroplasty. Because no single method has emerged as the optimal choice for all such fractures, it is important to understand which options are appropriate for each fracture pattern. Early classification systems focused on displacement as a major indication for either surgical or nonsurgical management. However, recent techniques and current implants have made surgical management preferable for most periprosthetic fractures. Classification based on fracture location can help guide such treatment. Generally, intramedullary nails are best for proximal fractures, fixed-angle devices for fractures originating at the component, and revision arthroplasty for very distal fractures or those with implant loosening. PMID- 14753794 TI - Exercise prescription. AB - Although many studies suggest that exercise provides various health benefits, the optimal dimensions of exercise (ie, frequency, intensity, duration, type) remain to be carefully defined. Research encompasses many areas, from activities of daily living to elite athletic performance. To properly prescribe a beneficial exercise program, a physician must have a basic understanding of the elements of exercise physiology and energy metabolism, as well as of the cardiovascular, respiratory, and musculoskeletal responses to exercise and training. Also, integration of specific programs for muscle strengthening with aerobic conditioning is important to achieve optimal results in patients with musculoskeletal conditions such as osteoarthritis and osteoporosis. PMID- 14753795 TI - Management of posttraumatic segmental bone defects. AB - Because of difficulty in managing posttraumatic segmental bone defects and the resultant poor outcomes, amputation historically was the preferred treatment. Massive cancellous bone autograft has been the principal alternative to amputation. Primary shortening or use of the adjacent fibula as a graft also has been used to attempt limb salvage. Of more recent methods of management, bone transport with distraction osteogenesis has been suggested as the leading option for defects of 2 to 10 cm, but problems include delayed union at the docking site and prolonged treatment time. Free vascularized bone transfer has been suggested as the leading option for defects of 5 to 12 cm, but hypertrophy of the graft is unreliable and late fracture, common. Bone graft substitutes continue to be developed, but they have not yet reached clinical efficacy for posttraumatic segmental bone defects. Although each of the new techniques has shown some limited success, complications remain common. PMID- 14753796 TI - Surgical treatment of congenital syndactyly of the hand. AB - Syndactyly is a congenital anomaly of the hand that is more common in males, is present bilaterally in 50% of affected patients, and often is associated with other musculoskeletal malformations or systemic syndromes. The goal of syndactyly release is to create a functional hand with the fewest surgical procedures while minimizing complications. For simple syndactyly, surgical reconstruction can begin at approximately 6 months, although many surgeons prefer to wait until the infant is 18 months old. Special situations, such as complex syndactyly and involvement of border digits, may warrant surgical intervention earlier than 6 months. Reconstruction of the web commissure is the most technically challenging part of the operation, followed by separation of the remaining digits. Full thickness skin grafting is almost always required for soft-tissue coverage. Complex syndactyly and syndactyly associated with other hand anomalies warrant special consideration. After reconstruction, patients should be examined periodically until they have achieved skeletal maturity because late complications such as web creep can occur. PMID- 14753797 TI - Coccygodynia: evaluation and management. AB - Coccygodynia is pain in the region of the coccyx. In most cases, abnormal mobility is seen on dynamic standing and seated radiographs, although the cause of pain is unknown in other patients. Bone scans and magnetic resonance imaging may show inflammation and edema, but neither technique is as accurate as dynamic radiography. Treatment for patients with severe pain should begin with injection of local anesthetic and corticosteroid into the painful segment. Coccygeal massage and stretching of the levator ani muscle can help. Coccygectomy is done only when nonsurgical treatment fails, which is infrequent. Coccygectomy usually is successful in carefully selected patients, with the best results in those with radiographically demonstrated abnormalities of coccygeal mobility. PMID- 14753798 TI - Advances in radionuclide therapeutics in orthopaedics. AB - Radiopharmaceuticals not only are used for diagnostic purposes but also increasingly in the treatment of many orthopaedic-related disorders. With the development of specific bone-seeking radiopharmaceuticals, the side effects of treatment are minimized, therapeutic effects are sustained, and concomitant use with other modalities may have synergistic effects. These new radiopharmaceuticals, such as strontium 89 and samarium 153-ethylene diamine tetramethylene phosphate, have been used as palliative treatment for patients with bone pain from osseous metastases. Excellent clinical responses with acceptable hematologic toxicity have been observed, and clinical results rival those of external beam radiation therapy. Radiosynovectomy has become a procedure of choice at many institutions to treat recurrent hemarthrosis and chronic synovitis in patients whose hemophilia is poorly controlled with medical management. Radiosynovectomy also remains a viable option to treat chronic synovitis secondary to inflammatory arthropathies, particularly rheumatoid arthritis. PMID- 14753799 TI - Unprecedented low Cu-N(azide)-Cu angles in end-on double azido bridged copper(II) complex. AB - A rare asymmetric end-on double azido-bridged copper(II) complex has been synthesized and characterized structurally and magnetically. The Cu-N(azide)-Cu angle in this complex is calculated to be 89.1 degrees. This is unusually low in comparison to the same angle in other end-on azido-bridged binuclear complexes reported so far. Though a strong ferromagnetic interaction between the metal centers is expected in the complex, the coupling has actually been found to be antiferromagnetic, instead. PMID- 14753800 TI - Formation of trinuclear palladium orthometalated complexes with unprecedented asymmetrical (mu 3-S)(mu 3-X) bridges (X = OH, SR, O2CR) from mu 2-hydroxo dimeric complexes and CS2. AB - Complexes [Pd(3)(mu(3)-S)(mu(3)-X)(L)(3)] (L = orthometalated imine), obtained by an unusual reaction of mu(2)-OH dimeric complexes and CS(2), are an unprecedented type of asymmetrical bridges between metallatriangles, which force an all-cis arrangement of the three orthometalated ligands relative to the metallatriangle. PMID- 14753801 TI - Encapsulation of cationic ruthenium complexes into a chiral self-assembled cage. AB - A chiral supramolecular assembly encapsulates the two cationic ruthenium sandwich complexes [CpRu(eta(6)-C(6)H(6))](+) and [CpRu(p-cymene)](+). The host-guest complexes K(11)[CpRu(eta(6)-C(6)H(6)) subset Ga(4)L(6)] (2) and K(11)[CpRu(p cymene) subset Ga(4)L(6)] (3) were characterized by one- and two-dimensional NMR techniques as well as by electrospray mass spectrometry. Encapsulation of the prochiral complex [CpRu(p-cymene)](+) by the chiral host renders enantiotopic protons diastereotopic as evidenced by (1)H NMR spectroscopy. PMID- 14753802 TI - Slow magnetic relaxation in a mixed-valence Mn(II/III) Complex: [MnII2(bispicen)2(mu 3-Cl)2Mn(III)(Cl4Cat)2Mn(III)(Cl4Cat)2(H2O)2] infinity. AB - A layered mixed-valence manganese complex, [Mn(II)(2)(bispicen)(2)(mu(3) Cl)(2)Mn(III)(Cl(4)Cat)(2)Mn(III)(Cl(4)Cat)(2)(H(2)O)(2)](infinity), is synthesized and characterized structurally. It displays a slow magnetic relaxation and hysteresis effect. PMID- 14753803 TI - [Cu(L)Mn(N3)2]n: the first complex containing both macrocyclic oxamido and alternate (mu-1,1 and mu-1,3) azido bridges. AB - A wattle-type heteropolynuclear complex [(CuL)Mn(N(3))(2)](n) (1) (H(2)L = 2,3 dioxo-5,6:15,16-dibenzo-1,4,8,13-tetraazacyclo-pentadeca-7,13-diene), which represents the first system with macrocyclic oxamido and alternate azido bridges, forming a one-dimensional chain structure, has been synthesized and characterized by a low temperature magnetic study. The chains are stacked with interchain hydrogen and lead to the structure of a three-dimensional network. PMID- 14753804 TI - Synthesis and characterization of N-heterocyclic carbene complexes of uranium(III). AB - Reaction of [(((Ad)ArO)(3)tacn)U(III)] (1) or [((Me(3)Si)(2)N)(3)U(III)] (3) with tetramethylimidazol-2-ylidene (Me(4)IMC:) yields novel N-heterocyclic carbene complexes [(((Ad)ArO)(3)tacn)U(III)(Me(4)IMC:)] (2) and [((Me(3)Si)(2)N)(3)U(III)(Me(4)IMC:)] (4). Uranium complexes 2 and 4 represent the first examples of compounds with an N-heterocyclic carbene ligand coordinated to a low-valent uranium center. The paramagnetic complexes 1, 2, and 4 were characterized by (1)H NMR, UV-vis-NIR, and EPR spectroscopy as well as SQUID magnetization measurements and X-ray diffraction analyses. DFT studies indicate a significant degree of pi-bonding in the U(III)-carbene entity. PMID- 14753805 TI - Calculated volume and energy profiles for water exchange on t2g6 rhodium(III) and iridium(III) hexaaquaions: conclusive evidence for an Ia mechanism. AB - An I(a) mechanism was assigned for water exchange on the hexaaquaions Rh(OH(2))(6)(3+) and Ir(OH(2))(6)(3+) on the basis of negative Delta V(++) experimental values (-4.2 and -5.7 cm(3) mol(-1), respectively). The use of Delta V(++) as a mechanistic criterion was open to debate primarily because Delta V(++) could be affected by extension or compression of the nonparticipating ligand bond lengths on going to the transition state of an exchange process. In this paper, volume and energy profiles for two distinct water exchange mechanisms (D and I(a)) have been computed using quantum chemical calculations which include hydration effects. The activation energy for Ir(OH(2))(6)(3+) is 32.2 kJ mol(-1) in favor of the I(a) mechanism (127.9 kJ mol(-1)), as opposed to a D pathway; the value for the I(a) mechanism being close to Delta H(++) and Delta G(++) experimental values (130.5 kJ mol(-1) and 129.9 kJ mol(-1) at 298 K, respectively). Volumes of activation, computed using Connolly surfaces and for the I(a) pathway (DeltaV(++)(calc) = -3.9 and -3.5 cm(3) mol(-1), respectively, for Rh(3+) and Ir(3+)), are in agreement with the experimental values. Further, it is demonstrated for both mechanisms that the contribution to the volume of activation due to the changes in bond lengths between Ir(III) and the spectator water molecules is negligible: -1.8 for the D, and -0.9 cm(3) mol(-1) for I(a) mechanism. This finding clarifies the debate about the interpretation of Delta V(++) and unequivocally confirms the occurrence of an I(a) mechanism with retention of configuration and a small a character for both Rh(III) and Ir(III) hexaaquaions. PMID- 14753806 TI - Mechanistic changeover for the water substitution on fac-[(CO)3Re(H2O)3]+ revealed by high-pressure NMR. AB - The complex formation in water between the stable tricarbonyltriaqua fac [(CO)(3)Re(H(2)O)(3)](+) (1) complex and N- and S-donor ligands has been studied by high-pressure (1)H NMR. Rate and equilibrium constants for the formation of [(CO)(3)Re(Pyz)(H(2)O)(2)](+), [(CO)(3)(H(2)O)(2)Re(mu Pyz)Re(H(2)O)(2)(CO)(3)](2+), [(CO)(3)Re(THT)(H(2)O)(2)](+), and [(CO)(3)Re(DMS)(n)()(H(2)O)(3-n)](+) (n = 1-3) (Pyz = pyrazine, THT = tetrahydrothiophene, DMS = dimethyl sulfide) have been determined and are in accord with previous results (Salignac, B.; Grundler, P. V.; Cayemittes, S.; Frey, U.; Scopelliti, R.; Merbach, A. E.; Hedinger, R.; Hegetschweiler, K.; Alberto, R.; Prinz, U.; Raabe, G.; Kolle, U.; Hall, S. Inorg. Chem. 2003, 42, 3516). The calculated interchange rate constant k(1)' (Eigen-Wilkins mechanism) increases from the hard O- and N-donors to the soft S-donors, as exemplified by the following series: TFA (trifluoroacetate) (k(1)' = 2.9 x 10(-3) s(-1)) < Br(-) < CH(3)CN < Pyz < THT < DMS < TU (thiourea) (k(1)' = 41.5 x 10(-3) s(-1)). On the other hand, values remain close to that of water exchange k(ex) on 1 (k(ex) = 6.3 x 10(-3) s(-1)). Thus, an I(d) mechanism was assigned, suggesting however the possibility of a slight deviation toward an associatively activated mechanism with the S-donor ligands. Activation volumes determined by high-pressure NMR, for Pyz as Delta V(++)(f,1) = +5.4 +/- 1.5, Delta V(++)(r,1) = +7.9 +/- 1.2 cm(3) mol(-)(1), for THT as Delta V(++)(f,1) = -6.6 +/- 1, Delta V(++)(r,1) = -6.2 +/- 1 cm(3) mol(-1), and for DMS as Delta V(++)(f,1) = -12 +/- 1, Delta V(++)(r,1) = 10 +/- 2 cm(3) mol(-1) revealed the ambivalent character of 1 toward water substitution. Hence, these findings are interpreted as a gradual changeover of the reaction mechanism from a dissociatively activated one (I(d)), with the hard O- and N-donor ligands, to an associatively activated one (I(a)), with the soft S donor ligands. PMID- 14753807 TI - New insights into the structural and dynamical features of lithium hexaoxometalates Li7MO6 (M = Nb, Ta, Sb, Bi). AB - We present a (re)investigation of the hexaoxometalates Li(8)MO(6) (M = Sn, Pb, Zr, Hf) and Li(7)MO(6) (M = Nb, Ta, Sb, Bi). Lithium motion and ionic conductivity in the hexaoxometalates were studied using impedance spectroscopy (for Li(7)MO(6), M = Sb, Bi, Ta) and (6)Li and (7)Li solid-state nuclear magnetic resonance (for Li(7)TaO(6)). The NMR data indicate a considerable exchange of Li among the tetrahedral and octahedral voids even at ambient temperature. In an investigation of the crystal structures using laboratory and synchrotron X-ray powder diffraction techniques, the structures of Li(7)TaO(6), Li(7)NbO(6), and Li(7)SbO(6) could be solved and refined. All three reveal a triclinic metric (Li(7)SbO(6), triclinic, P1, a = 5.38503(6) A, b = 5.89164(7) A, c = 5.43074(6) A, alpha = 117.2210(6) degrees, beta = 119.6311(6) degrees, gamma = 63.2520(7) degrees, V = 127.454(3) A(3), Z = 1; Li(7)NbO(6), triclinic, P1, a = 5.37932(9) A, b = 5.91942(11) A, c = 5.37922(9) A, alpha = 117.0033(9) degrees, beta = 119.6023(7) degrees, gamma = 63.2570(9) degrees, V = 126.938(4) A(3), Z = 1; Li(7)TaO(6), triclinic, P1, a = 5.38486(2) A, b = 5.92014(3) A, c = 5.38551(2) A, alpha = 117.0108(2) degrees, beta = 119.6132(2) degrees, gamma = 63.2492(2) degrees, V = 127.208(1) A(3), Z = 1. PMID- 14753808 TI - On the noble-gas-induced intersystem crossing for the CUO molecule: experimental and theoretical investigations of CUO(Ng)n (Ng = Ar, Kr, Xe; n = 1, 2, 3, 4) complexes in solid neon. AB - Uranium atoms excited by laser ablation react with CO in excess neon to produce the novel CUO molecule, which forms distinct Ng complexes (Ng = Ar, Kr, Xe) when the heavier noble gases are added. The CUO(Ng) complexes are identified through CO isotopic and Ng substitution on the neon matrix infrared spectra and by comparison to DFT frequency calculations. The U-C and U-O stretching frequencies of CUO(Ng) complexes are slightly red-shifted from frequencies for the (1)Sigma(+) CUO ground state, which identifies singlet ground state CUO(Ng) complexes. In solid neon the CUO molecule is also a complex CUO(Ne)(n), and the CUO(Ne)(n-1)(Ng) complexes are likewise specified. The next singlet CUO(Ne)(x)(Ng)(2) complexes in excess neon follow in like manner. However, the higher CUO(Ne)(x)(Ng)(n) complex (n = 3, 4) stretching modes approach pure argon matrix CUO(Ar)(n) values and isotopic behavior, which are characterized as triplet ground state complexes by DFT frequency calculations. This work suggests that the singlet-triplet crossing occurs with 3 Ar, 3 Kr, or 4 Xe and a balance of Ne atoms coordinated to CUO in the neon matrix host. PMID- 14753809 TI - Characterization of the thermally induced topochemical solid-state transformation of NH4[N(CN)2] into NCN[double bond]C(NH2)2 by means of X-ray and neutron diffraction as well as Raman and solid-state NMR spectroscopy. AB - The mechanism of the solid-solid transformation of NH(4)[N(CN)(2)] into NCN[double bond]C(NH(2))(2), which represents the isolobal analogue of Wohler's historic conversion of ammonium cyanate into urea, has been investigated by temperature-dependent single-crystal and powder X-ray diffraction, neutron powder diffraction, and Raman and solid-state NMR spectroscopy as well as thermoanalytical measurements. The transformation of the ionic dicyanamide into its molecular isomer upon controlled thermal treatment was found to proceed topochemically in the solid state with little molecular motion, giving rise to a single-crystal to single-crystal transformation which manifests itself by a defined metric relation between the unit cells of the two isomers. The exothermic phase transition is thermally activated and was observed to commence at temperatures > or =80 degrees C. The pronounced temperature dependence of the onset of the transformation may be assessed as an indication for the metastability of ammonium dicyanamide at elevated temperatures. Thermal analyses reveal a decrease in the reaction enthalpy (56-13 kJ mol(-1)) at higher heating rates and an average mass loss of 10% gaseous ammonia. Evidence was found for crucial mechanistic steps of the transformation, which is likely to proceed via proton transfer from the ammonium ion to one of the terminal nitrogen atoms of the anion. The protonation is followed by nucleophilic attack of the in situ generated ammonia at the electrophilic nitrile carbon. The proposed mechanistic pathway is based on the results of combined Raman and solid-state NMR spectroscopic as well as neutron powder diffraction measurements. PMID- 14753810 TI - Biologically active platinum complexes containing 8-thiotheophylline and 8 (methylthio)theophylline. AB - Complexes [Pt(mu-N,S-8-TT)(PPh(3))(2)](2) (1), [Pt(mu-S,N-8-TT)(PTA)(2)](2) (2), [Pt(8-TTH)(terpy)]BF(4) (3), cis-[PtCl(8-MTT)(PPh(3))(2)] (4), cis-[Pt(8 MTT)(2)(PPh(3))(2)] (5), cis-[Pt(8-MTT)(8-TTH)(PPh(3))(2)] (6), cis-[PtCl(8 MTT)(PTA)(2)] (7), cis-[Pt(8-MTT)(2)(PTA)(2)] (8), and trans-[Pt(8 MTT)(2)(py)(2)] (9) (8-TTH(2) = 8-thiotheophylline; 8-MTTH = 8 (methylthio)theophylline; PTA = 1,3,5-triaza-7-phosphaadamantane) are presented and studied by IR and multinuclear ((1)H, (31)P[(1)H]) NMR spectroscopy. The solid-state structure of 4 and 9 has been authenticated by X-ray crystallography. Growth inhibition of the cancer cells T2 and SKOV3 induced by the above new thiopurine platinum complexes has been investigated. The activity shown by complexes 4 and 9 was comparable with cisplatin on T2. Remarkably, 4 and 9 displayed also a valuable activity on cisplatin-resistant SKOV3 cancer cells. PMID- 14753811 TI - Structural study of the T#2-LixCoO2 (0.52 < x < or = 0.72) phase. AB - The metastable O2-LiCoO(2) phase undergoes several reversible phase transitions upon lithium deintercalation. The first transition leads to an unusual oxygen stacking in such layered compounds. This stacking is found to be stable for 0.52 < x < or = 0.72 in Li(x)()CoO(2) and is called T(#)2. We studied this phase from a structural viewpoint using X-ray and neutron diffraction (ab initio method). The new stacking derives from the O2 one by gliding every second CoO(2) slab by (1/3, 1/6, 0). The lithium ions are found to occupy very distorted tetrahedral sites in this structure. We also discuss the possibility of this T(#)2 phase to exhibit stacking faults, whose amount depends on the method used to prepare this deintercalated phase. PMID- 14753812 TI - Enzymatic mechanism of Fe-only hydrogenase: density functional study on H-H making/breaking at the diiron cluster with concerted proton and electron transfers. AB - The mechanism of the enzymatic hydrogen bond forming/breaking (2H(+) + 2e<==>H(2)) and the plausible charge and spin states of the catalytic diiron subcluster [FeFe](H) of the H cluster in Fe-only hydrogenases are probed computationally by the density functional theory. It is found that the active center [FeFe](H) can be rationally simulated as [[H](CH(3)S)(CO)(CN( ))Fe(p)(CO(b))(mu-SRS)Fe(d)(CO)(CN(-))L], where the monovalence [H] stands for the [4Fe4S](H)(2+) subcluster bridged to the [FeFe](H) moiety, (CH(3)S) represents a Cys-S, and (CO(b)) represents a bridging CO. L could be a CO, H(2)O, H(-), H(2), or a vacant coordination site on Fe(d). Model structures of possible redox states are optimized and compared with the X-ray crystallographic structures and FTIR experimental data. On the basis of the optimal structures, we study the most favorable path of concerted proton transfer and electron transfer in H(2)-forming/breaking reactions at [FeFe](H). Previous mechanisms derived from quantum chemical computations of Fe-only hydrogenases (Cao, Z.; Hall, M. B. J. Am. Chem. Soc. 2001, 123, 3734; Fan, H.; Hall, M. B. J. Am. Chem. Soc. 2001, 123, 3828) involved an unidentified bridging residue (mu-SRS), which is either a propanedithiolate or dithiomethylamine. Our proposed mechanism, however, does not require such a ligand but makes use of a shuttle of oxidation states of the iron atoms and a reaction site between the two iron atoms. Therefore, the hydride H(b)(-) (bridged to Fe(p) and Fe(d)) and eta(2)-H(2) at Fe(p) or Fe(d) most possibly play key roles in the dihydrogen reversible oxidation at the [FeFe](H) active center. This suggested way of H(2) formation/splitting is reminiscent of the mechanism of [NiFe] hydrogenases and therefore would unify the mechanisms of the two related enzymes. PMID- 14753813 TI - Coordination polymers assembled from angular dipyridyl ligands and CuII, CdII, CoII salts: crystal structures and properties. AB - Six new metal-organic coordination networks based on linking unit 2,5-bis(4 pyridyl)-1,3,4-thiadiazole (L(1)) or 2,5-bis(3-pyridyl)-1,3,4-oxadiazole (L(3)) and inorganic Cu(II), Cd(II), and Co(II) salts have been prepared and structurally characterized by single-crystal X-ray analysis. Using L(1) to react with three different Cu(II) salts, Cu(OAc)(2).H(2)O, Cu(NO(3))(2).3H(2)O, and CuSO(4).5H(2)O, respectively, two different one-dimensional (1-D) coordination polymers, [[Cu(2)L(1)(mu-OAc)(4)](CHCl(3))(2)](n) (1) [triclinic, space group P1, a = 7.416(3) A, b = 8.207(3) A, c = 14.137(5) A, alpha = 100.333(7) degrees, beta = 105.013(6) degrees, gamma = 94.547(6) degrees, Z = 1] and [[CuL(1)(NO(3))(2)](CHCl(3))(0.5)](n) (2) [monoclinic, space group C2/c, a = 28.070(8) A, b = 9.289(3) A, c = 15.235(4) A, beta = 113.537(5) degrees, Z = 8], and a chiral 3-D open framework, [[CuL(1)(H(2)O)(SO(4))](H(2)O)(2)](n) (3) [orthorhombic, space group P2(1)2(1)2(1), a = 5.509(2) A, b = 10.545(4) A, c = 29.399(11) A, Z = 4], were obtained. Reaction of L(1) and Cd(ClO(4))(2).6H(2)O or Co(ClO(4))(2).6H(2)O, in the presence of NH(4)SCN, yielded another 3-D open framework, [[CdL(1)(NCS)(2)](CH(3)OH)(1.5)](n) (4) [monoclinic, space group C2/c, a = 28.408(10) A, b = 9.997(5) A, c = 7.358(4) A, beta = 99.013(8) degrees, Z = 4], or a 2-D network, [[Co(L(1)())(2)(NCS)(2)](H(2)O)(2.5)](n) (5) [orthorhombic, space group Pnna, a = 22.210(5) A, b = 12.899(3) A, c = 20.232(4) A, Z = 4]. When L(1) was replaced by L(3) to react with Co(ClO(4))(2).6H(2)O and NH(4)SCN, another 2-D coordination polymer, [Co(L(3))(2)(NCS)(2)](n) (6) [monoclinic, space group P2(1)/c, a = 8.120(3) A, b = 9.829(4) A, c = 17.453(6) A, beta = 103.307(6) degrees, Z = 2], was constructed. These results indicate that the nature of the ligands, metal centers, or counteranions plays the critical role in construction of these novel coordination polymers. The interesting porous natures of two 3-D open frameworks 3 and 4 were investigated by TGA and XPRD techniques, and the magnetic properties of the Cu(II) and Co(II) complexes were studied by variable temperature magnetic susceptibility and magnetization measurements. PMID- 14753814 TI - Monomeric, tetrameric, and polymeric copper di-tert-butyl phosphate complexes containing pyridine ancillary ligands. AB - The reaction of di-tert-butyl phosphate (((t)BuO)(2)P(O)(OH), dtbp-H) with copper acetate in the presence of pyridine (py) and 2,4,6-trimethylpyridine (collidine) has been investigated. Copper acetate reacts with dtbp-H in a reaction medium containing pyridine, DMSO, THF, and CH(3)OH to yield a one-dimensional polymeric complex [Cu(dtbp)(2)(py)(2)(mu-OH(2))](n) (1) as blue hollow crystalline tubes. The copper atoms in 1 are octahedral and are surrounded by two terminal phosphate ligands, two pyridine molecules, and two bridging water molecules. The mu-OH(2) ligands that are present along the elongated Jahn-Teller axis are responsible for the formation of the one-dimensional polymeric structure. Recrystallization of 1 in a DMSO/THF/CH(3)OH mixture results in the reorganization of the polymer and its conversion to a more stable tetranuclear copper cluster [Cu(4)(mu(3) OH)(2)(dtbp)(6)(py)(2)] (2) in about 60% yield. The molecular structure of 2 is made up of a tetranuclear core [Cu(4)(mu(3)-OH)(2)] which is surrounded by six bidentate bridging dtbp ligands. While two of the copper atoms are pentacoordinate with a tbp geometry, the other two copper atoms exhibit a pseudooctahedral geometry with five normal Cu-O bonds and an elongated Cu-O linkage. The pentacoordinate copper centers bear an axial pyridine ligand. The short Cu.Cu nonbonded distances in the tetranuclear core of 2 lead to magnetic ordering at low temperature with an antiferromagnetic coupling at approximately 20 K (J(P) = -44 cm(-1), J(c) = -66 cm(-1), g = 2.25, and rho = 0.8%). When the reaction between di-tert-butyl phosphate (dtbp-H) and copper acetate was carried out in the presence of collidine, large dark-blue crystals of monomeric copper complex [Cu(dtbp)(2)(collidine)(2)] (3) formed as the only product. A single crystal X-ray diffraction study of 3 reveals a slightly distorted square-planar geometry around the copper atom. Thermogravimetric analysis of 1-3 revealed a facile decomposition of the coordinated ligands and dtbp to produce a copper phosphate material around 500 degrees C. An independent solid-state thermolysis of all the three complexes in bulk at 500-510 degrees C for 2 days produced copper pyrophosphate Cu(2)P(2)O(7) along with small quantities of Cu(PO(3))(2) as revealed by DR-UV spectroscopic and PXRD studies. PMID- 14753815 TI - Novel double layer salts of copper(I) bromide with N-substituted ethylenediammonium cations. AB - The structures of three hybrid organic/inorganic halometalate salts are reported, and the layer structures developed are contrasted. Crystal structures of the isostructural N-methylethylenediammonium (MEDA(2+)) and N-ethylethylenediammonium (EEDA(2+)) salts of copper(I) bromide are both triclinic, space group P1, with lattice constants a = 6.284(7), b = 7.842(6), and c = 12.03(1) A, alpha = 84.84(3), beta = 83.08(2), and gamma = 88.00(3) degrees, and V = 586(1) A(3) with Z = 2 for (MEDA)Cu(2)Br(4) while (EEDA)Cu(2)Br(4) has lattice constants a = 6.27(2), b = 7.78(2), and c = 13.12(3) A, alpha = 84.69(4), beta = 78.18(3), and gamma = 88.17(7) degrees, and V = 623(3) A(3) with Z = 2. The dominant inorganic feature in both salts is anionic (CuBr(2))(n)(n-) chains of edge-shared CuBr(4) tetrahedra. The diammonium cations hydrogen bond these chains together into a unique double layer structure. For comparison purposes, the crystal structure of (CHA)PbBr(3) (CHA(+) = cyclohexylammonium) is reported (monoclinic, space group P2(1)/n, a = 8.088(2) A, b = 7.912(2) A, and c = 19.572(4) A, beta = 96.98(4) degrees, and V = 1243.2(4) A(3) with Z = 4). This contains (PbBr(3))(n)(n-) halometalate chains, this time of face-shared PbBr(6) octahedra. However, here the organic cations tie the chains together into the more common single layer structure. PMID- 14753816 TI - Hydrothermal synthesis, structure, and magnetic properties of the mixed-valent Np(IV)/Np(V) Selenite Np(NpO2)2(SeO3)3. AB - The reaction of NpO(2) with SeO(2) in the presence of CsCl at 180 degrees C results in the formation of Np(NpO(2))(2)(SeO(3))(3) (1). The structure of 1 consists of three crystallographically unique Np centers with three different coordination environments in two different oxidation states. Np(1) is found in a neptunyl(V), O[double bond]Np[double bond]O(+), unit that is further ligated in the equatorial plane by three chelating SeO(3)(2-) anions to create a hexagonal bipyramidal NpO(8) unit. A second neptunyl(V) cation also occurs for Np(2); it is bound by four bridging selenite anions and by the oxo atom from the Np(1) neptunyl cation to form a pentagonal bipyramidal, NpO(7), unit. The third neptunium center, Np(3), which contains Np(IV), is found in a distorted NpO(8) dodecahedron. Np(3) is bound by five bridging selenite anions and by three neptunyl units via cation-cation interactions. The NpO(7) pentagonal bipyramids and NpO(8) hexagonal bipyramids share both corners and edges. Both of these polyhedra share corners via cation-cation interactions with the NpO(8) dodecahedra creating a three-dimensional structure with small channels that house the stereochemically active lone pair of electrons on the selenite anions. Magnetic susceptibility data follow Curie-Weiss behavior over the entire temperature range measured (5 < or = T < or = 320 K). The effective moment, mu(eff) = 2.28 mu(B), which represents an average over the three crystallographically inequivalent Np atoms, is within the expected range of values. There is no evidence of long-range ordering of the Np moments at temperatures down to 5 K, consistent with the negligible Weiss constant determined from fitting the susceptibility data. Crystallographic data: 1, orthorhombic, space group Pbca, a = 10.6216(5), b = 11.9695(6), and c = 17.8084(8) A and Z = 8 (T = 193 K). PMID- 14753817 TI - Synthesis, structure, and characterization of two new layered mixed-metal phosphates, BaTeMO4(PO4) (M = Nb5+ or Ta5+. AB - Two new isostructural mixed-metal phosphates, BaTeMO(4)(PO(4)) (M = Nb(5+) or Ta(5+)), have been synthesized as bulk phase powders and single crystals by standard solid-state techniques using BaCO(3), TeO(2), Nb(2)O(5) (or Ta(2)O(5)), and NH(4)H(2)PO(4) as reagents. The materials have novel layered crystal structures consisting of [M(5+)O(6/2)](-) corner-sharing octahedral chains that are connected to [Te(4+)O(4/2)](0) polyhedra and [P(5+)O(2/1)O(2/2)](-) tetrahedra. The Ba(2+) cations reside between the layers and maintain charge balance. The Te(4+) cations are in asymmetric coordination environments attributable to their lone pairs. The Nb(5+) distorts along the local C(4) direction of its octahedron resulting in a "short-long-short-long" Nb-O-Nb bond motif. The Nb(5+) cation displaces away from the oxide ligands that are bonded to Te(4+) or P(5+) cations, attributable to the structural rigidity of the TeO(4) and PO(4) polyhedra. Thus, the TeO(4) and PO(4) polyhedra support and reinforce the intraoctahedral distortion observed within the NbO(6) octahedra. Infrared and Raman spectroscopy, thermogravimetric analysis, and ion-exchange experiments are also presented. Crystal data: BaTeNbO(4)(PO(4)), orthorhombic, space group Pbca (No. 61), with a = 6.7351(9) A, b = 7.5540(10) A, c = 27.455(4) A, V = 1396.8(3) A(3), and Z = 8; BaTeTaO(4)(PO(4)), orthorhombic, space group Pbca (No. 61), with a = 6.734(2) A, b = 7.565(3) A, c = 27.435(9) A, V = 1372.6(8) A(3), and Z = 8. PMID- 14753818 TI - Problem of a wide variety of products in the Cu(hfac)2-nitroxide system. AB - The stereochemically flexible Cu(hfac)(2) metal-ligand system when combined with polyfunctional nitroxides leads to a variety of solids with varying structure and composition. While investigating the products of Cu(hfac)(2) interaction with spin-labeled pyrazole 4,4,5,5-tetramethyl-2-(1-methyl-1H-pyrazol-4-yl) imidazoline-3-oxide-1-oxyl, we have isolated a family of (12) heterospin compounds differing in structure and composition in the solid state. In synthetic systems, these compounds often cocrystallize and must be separated mechanically. It is also shown that minor variation of the structure of the solid heterospin complex can substantially change the magnetic properties of compounds. PMID- 14753819 TI - Palladium(II) complexes of bowls, pinwheels, cages, and N,C,N-pincers of starburst ligands 1,3,5-Tris(di-2-pyridylamino)benzene and 2,4,6-Tris(di-2 pyridylamino)-1,3,5-triazene. AB - The reactions of Pd(II) ions with starburst ligands 1,3,5-tris(di-2 pyridylamino)benzene (tdab) and 2,4,6-tris(di-2-pyridylamino)-1,3,5-triazene (tdat) have been investigated. Complexes with the Pd:tdab (or tdat) ratio being 1:1 and 3:1 have been isolated and characterized. The structures of five new Pd(II) complexes containing the starburst ligands have been determined by X-ray diffraction analyses, which include chelate compounds [PdCl(2)(tdab)], 1, [(PdCl(2))(3)(tdab)], 2, [(Pd(OAc)(2))(3)(tdab)], 4, and [(Pd(OAc)(2))(3)(tdat)], 5, and a cyclometalated compound [Pd(OAc)(NCN-tdab)], 3. The Pd(II) ion in the 1:1 compound 1 is chelated by two pyridyl groups. Similarly, each Pd(II) center in the 3:1 compounds 2, 4, and 5 is chelated by two pyridyl groups. However, these three compounds display distinct structural features: 2 adopts a "bowl shaped" structure, 4 has a "pinwheel"-like structure, and 5 has a "up-and-down" structure. Compounds 4 and 5 were examined in solution by variable-temperature (1)H NMR, which revealed that both compounds retain the "pinwheel" and the "up and-down" structure, respectively. The observed structural preference by 4 and 5 is attributed to both electronic and steric factors. PMID- 14753820 TI - Insertion reactions of [M(SR)3(PMe2Ph)2] with CS2 (M = Ru, Os; R = C6F4H-4, C6F5). X-ray structures of [Ru(S2CSC6F4H-4)2(PMe2Ph)2], trans-thiolates [M(SR)2(S2CSR)(PMe2Ph)2] (M = Ru; R = C6F5 and M = Os; R = C6F4H-4), and trans thiolate-phosphine [Os(SC6F5)2(S2CSC6F5)(PMe2Ph)2]. AB - Reactions of [M(SR)(3)(PMe(2)Ph)(2)] (M = Ru, Os; R = C(6)F(4)H-4, C(6)F(5)) with CS(2) in acetone afford [Ru(S(2)CSR)(2)(PMe(2)Ph)(2)] (R = C(6)F(4)H-4, 1; C(6)F(5), 3) and trans-thiolates [Ru(SR)(2)(S(2)CSR)(PMe(2)Ph)(2)] (R = C(6)F(4)H 4, 2; C(6)F(5), 4) or the isomers trans-thiolates [Os(SR)(2)(S(2)CSR)(PMe(2)Ph)(2)] (R = C(6)F(4)H-4, 5; C(6)F(5), 7) and trans thiolate-phosphine [Os(SR)(2)(S(2)CSR)(PMe(2)Ph)(2)] (R = C(6)F(4)H-4, 6; C(6)F(5), 8) through processes involving CS(2) insertion into M-SR bonds. The ruthenium(III) complexes [Ru(SR)(3)(PMe(2)Ph)(2)] react with CS(2) to give the diamagnetic thiolate-thioxanthato ruthenium(II) and the paramagnetic ruthenium(III) complexes while osmium(III) complexes [Os(SR)(3)(PMe(2)Ph)(2)] react to give the paramagnetic thiolate-thioxanthato osmium(III) isomers. The single-crystal X-ray diffraction studies of 1, 4, 5, and 8 show distorted octahedral structures. (31)P [(1)H] and (19)F NMR studies show that the solution structures of 1 and 3 are consistent with the solid-state structure of 1. PMID- 14753821 TI - Insertion reactions of trans-Mo(dmpe)2(H)(NO) with imines. AB - The insertion chemistry of the hydride complex trans-Mo(dmpe)(2)(H)(NO) (1) (dmpe = bis(dimethylphosphino)ethane) with imines has been investigated. It was found that disubstituted aromatic imines RCH[double bond]NR' (R, R' = Ar) insert into the Mo-H bond of 1, while a series of various mono- and other disubstituted imines do not react. The insertion products trans-Mo(dmpe)(2)(NO)[NR'(CH(2)R)] (R = R' = Ph (2); R = Cp(2)Fe, R' = Ph (3); R = Ph, R' = Cp(2)Fe (4); R = 1 naphthyl, R' = Ph (5)) have been isolated and fully characterized by elemental analysis, IR and NMR spectroscopy, and mass spectrometry. The imine PhCH[double bond]NC(10)H(7) (C(10)H(7) = 1-naphthyl) reacted with 1 establishing an equilibrium to produce the nonisolable complex trans Mo(dmpe)(2)(NO)[NC(10)H(7)(CH(2)Ph)] (6). The equilibrium constant for this reaction has been derived from VT-NMR measurements, and the Delta H and Delta S values of this reaction were calculated to be -48.8 +/- 0.4 kJ.mol(-1) and -33 +/ 1 J.K(-1).mol(-1) reflecting a mild exothermic process and its associative nature. Single-crystal X-ray diffraction analyses were carried out on 2-5. PMID- 14753822 TI - Evidence for the blue 10 pi S62+ dication in solutions of S8(AsF6)2: a computational study including solvation energies. AB - The energetics of dissociation reactions of S(8)(2+) into stoichiometric mixtures of S(n)(+), n = 2-7, and S(m)(2+), m = 3, 4, 6, 10, were investigated by the B3PW91 method [6-311+G(3df)//6-311+G] in the gas phase and in solution, with solvation energies calculated using the SCIPCM model and in some cases also the COSMO model [B3PW91/6-311+G*, dielectric constants 2-30, 83, 110]. UV-vis spectra of all species were calculated at the CIS/6-311G(2df) level and for S(4)(2+) and S(6)(2+) also at the TD-DFT level (BP86/SV(P)). Standard enthalpies of formation at 298 K were derived for S(3)(2+) (2538 kJ/mol), S(6)(2+) (2238 kJ/mol), and S(10)(2+) (2146 kJ/mol). A comparison of the observed and calculated UV-vis spectra based on our calculated thermochemical data in solution suggests that, in the absence of traces of facilitating agent (such as dibromine Br(2)), S(8)(2+) dissociates in dilute SO(2) solution giving an equilibrium mixture of ca. 0.5S(6)(2+) and S(5)(+) (K approximately 8.0) while in the more polar HSO(3)F some S(8)(2+) remains (K approximately 0.4). According to our calculations, the blue color of this solution is likely due to the pi-pi transition of the previously unknown 10 pi S(6)(2+) dication, and the previously assigned S(5)(+) is a less important contributor. Although not strictly planar, S(6)(2+) may be viewed as a 10 pi electron Huckel-aromatic ring containing a thermodynamically stable 3p(pi)-3p(pi) bond [d(S-S) = 2.028 A; tau(S-S-S-S) = 47.6 degrees ]. The computations imply that the new radical cation S(4)(+) may be present in sulfur dioxide solutions given on reaction of sulfur oxidized by AsF(5) in the presence of a facilitating agent. The standard enthalpy of formation of S(6)(AsF(6))(2)(s) was estimated as -3103 kJ/mol, and the disproportionation enthalpy of 2S(6)(AsF(6))(2)(s) to S(8)(AsF(6))(2)(s) and S(4)(AsF(6))(2)(s) as exothermic by 6-17 kJ/mol. The final preference of the observed disproportionation products is due to the inclusion of solvent molecules, e.g., AsF(3), that additionally favors the disproportionation of 2S(6)(AsF(6))(2)(s) into S(8)(AsF(6))(2)(s) and S(4)(AsF(6))(2)(AsF(3))(s) by 144 kJ/mol. PMID- 14753823 TI - Electrochemical and spectroelectrochemical characterization of Ru(2)(4+) and Ru(2)(3+) complexes under a CO atmosphere. AB - Eleven different Ru(2)(4+) and Ru(2)(3+) derivatives are characterized by thin layer FTIR and UV-visible spectroelectrochemistry under a CO atmosphere. These compounds, which were in-situ electrogenerated from substituted anilinopyridine complexes with a Ru(2)(5+) core, are represented as Ru(2)(L)(4)Cl where L = 2 CH(3)ap, ap, 2-Fap, 2,3-F(2)ap, 2,4-F(2)ap, 2,5-F(2)ap, 3,4-F(2)ap, 3,5-F(2)ap, 2,4,6-F(3)ap, or F(5)ap. The Ru(2)(5+) complexes do not axially bind CO while mono- and bis-CO axial adducts are formed for the Ru(2)(4+) and Ru(2)(3+) derivatives, respectively. Six of the eleven investigated compounds exist in a (4,0) isomeric form while five adopt a (3,1) geometric conformation. These two series of compounds thus provide a large enough number of derivatives to examine trends and differences in the spectroscopic data of the two types of isomers in their lower Ru(2)(4+) and Ru(2)(3+) oxidation states. UV-visible spectra of the Ru(2)(4+) derivatives and IR spectra of the Ru(2)(3+) complexes under CO are both isomer dependent, thus suggesting that these data can be used to reliably predict the isomeric form, i.e., (3,1) or (4,0), of diruthenium complexes containing four unsymmetrical substituted anilinopyridinate bridging ligands; this was confirmed by X-ray crystallographic data for seven compounds whose structures were available. PMID- 14753824 TI - Self-assembly of cuII and niII [2 x 2] grid complexes and a binuclear CuII complex with a new semiflexible substituted pyrazine ligand: multiple anion encapsulation and magnetic properties. AB - With the new substituted pyrazine ligand pyrazine-2,3-dicarboxylic acid bis[(pyridin-2-ylmethyl)amide], H(2)L, a binuclear complex [Cu(2)(LH)(Cl(3))(H(2)O)].H(2)O (1) and two [2 x 2]G grid complexes, [[Cu(4)(LH)(4)](ClO(4))(4)].5CH(3)OH.4H(2)O (2) and [[Ni(4)(LH)(4)]Cl(4)].5CH(3)CN.13H(2)O (3), have been synthesized and characterized spectroscopically and crystallographically. The ligand H(2)L crystallized in the triclinic space group P1, with a = 4.9882(7) A, b = 12.079(2) A, c = 14.454(2) A, alpha = 107.08(2) degrees, beta = 98.61(2) degrees, gamma = 97.54(2) degrees, V = 808.8(2) A(3), Z = 2, R1 = 0.0747, and R(w) = 0.1829 for 1319 observed reflections [I > 2 sigma(I)]. The molecule is L-shaped with a strong intramolecular bifurcated hydrogen bond in half of the molecule. In the crystal the molecules are linked by an intermolecular hydrogen bond to form a 1D polymer. The binuclear complex [Cu(2)(LH)(Cl(3))(H(2)O)].H(2)O (1) crystallized in the monoclinic space group P2(1)/a, with a = 8.6859(7) A, b = 28.060(2) A, c = 9.5334(9) A, beta = 107.89(1) degrees, V = 2211.2(3) A(3), Z = 4, R1 = 0.039, and R(w) = 0.097 for 1408 observed reflections [I > 2 sigma(I)]. There are two independent copper atoms both having square pyramidal geometry. Both coordinate to a pyrazine, a pyridine, and an amide N atom. Two chlorines complete the coordination sphere of one of the copper atoms, while one chlorine atom and a water molecule complete the coordination sphere of the other. The copper(II) [2 x 2] grid complex [[Cu(4)(LH)(4)](ClO(4))(4)].5CH(3)OH.4H(2)O (2) crystallized in the triclinic space group P1, with a = 17.1515(14) A, b = 17.7507(13) A, c = 19.3333(15) A, alpha = 67.34(1) degrees, beta = 69.79(1) degrees, gamma = 71.50(1) degrees, V = 4980.3(7) A(3), Z = 2, R1 = 0.083, and R(w) = 0.207 for 5532 observed reflections [I > 2 sigma(I)]. The four Cu(II) atoms are octahedrally coordinated by two pyrazine, two pyridine, and two amide N atoms and occupy the corners of a [2 x 2] grid with edge lengths, Cu...Cu, varying from 7.01 to 7.39 A. The nickel(II) [2 x 2] grid complex [[Ni(4)(LH)(4)]Cl(4)].5CH(3)CN.13H(2)O (3) crystallized in the monoclinic space group C2/c, with a = 16.3388(10) A, b = 29.754(2) A, c = 20.857(1) A, beta = 101.845(1) degrees, V = 9923.6(12) A(3), Z = 4, R1 = 0.050, and wR2 = 0.101 for 3391 observed reflections [I > 2 sigma(I)]. Here the complex possesses C(2) symmetry and again each metal atom is octahedrally coordinated to two pyrazine, two pyridine, and two amide N atoms. They occupy the corners of a [2 x 2] grid with an average edge length, Ni.Ni, of 6.97 A. Of the four anions (ClO(4)(-)'s in 2 and Cl(-)'s in 3) required to equilibrate the charges in the grid complexes, two are encapsulated, one above and one below the plane of the four metal atoms. The remaining two anions are located between the "wings" of the ligands. Magnetic susceptibility measurements indicate that the binuclear complex 1 is antiferromagnetic, with a J value of -15.07 cm(-1). This is larger than the J values found for the Cu(II) (2) and Ni(II) (3) grid complexes, which were -5.87 and -2.64 cm(-1), respectively. DFT calculations have been carried out to explain the difference in the J values found for complexes 1 and 2. PMID- 14753825 TI - Oxidation in nonclassical organolanthanide chemistry: synthesis, characterization, and X-ray crystal structures of cerium(III) and -(IV) amides. AB - [Ce(NR(2))(3)] (R = SiMe(3)) with TeCl(4) in tetrahydrofuran solution gave a mixture of two major products in a combined yield of ca. 50% based on available metal: (i) the Ce(IV) amide [CeCl(NR(2))(3)] (1), which was isolated as purple needles and identified on the basis of (1)H NMR and mass spectra, microanalysis, and a single-crystal X-ray analysis [C(18)H(54)CeClN(3)Si(6), rhombohedral, R3c (No. 161), a = b = 18.4508(7) A, c = 16.8934(7) A, Z = 6]; (ii) unstable [[Ce(NR(2))(2)(mu-Cl)(thf)](2)] (2), as colorless blocks [C(32)H(88)Ce(2)Cl(2)N(4)O(2)Si(8), monoclinic, P2(1)/n (No. 14), a = 14.506(3) A, b = 13.065(3) A, c = 16.779(3) A, beta = 113.789(12) degrees, Z = 2], which readily disproportionated in solution. In toluene solution, the product 1 was obtained exclusively. The same cerium(III) amide starting material was oxidized by PBr(2)Ph(3) in diethyl ether solution to give purple [CeBr(NR(2))(3)] (3) [C(18)H(54)BrCeN(3)Si(6), rhombohedral, R3c (No. 161), a = b = 18.4113(12) A, c = 16.9631(17) A, Z = 6], along with presumed [CeBr(3)(OEt(2))(n)()], which has not been characterized but with thf, by displacement of the ether ligands, gave [CeBr(3)(thf)(4)] (4) [C(16)H(32)Br(3)CeO(4), triclinic, P1 (No. 2), a = 8.2536(7) A, b = 9.4157(5) A, c = 15.5935(14) A, alpha = 79.009(5), beta = 87.290(3) degrees, gamma = 74.835(5) degrees, Z = 2). TeBr(4) reacted with [Ce(NR(2))(3)] in thf to give small amounts of 3; the major product (although only formed in 15% yield) was monomeric [CeBr(2)(NR(2))(thf)(3)] (5) [C(18)H(42)Br(2)CeNO(3)Si(2), monoclinic, P2(1)/c (No. 14), a = 14.9421(4) A, b = 11.8134(5) A, c = 15.8317(7) A, alpha = gamma = 120 degrees, beta = 92.185(3) degrees, Z = 4]. PMID- 14753826 TI - The mechanism of NO formation from the decomposition of dialkylamino diazeniumdiolates: density functional theory and CBS-QB3 predictions. AB - The mechanism of decomposition of dialkylamino diazeniumdiolate salts, R(2)N[NONO](-)M(+), to form nitric oxide, NO, was explored using theoretical methods. B3LYP/6-311+G(d) density functional theory calculations gave the optimized geometries, and energetics were further evaluated with the high accuracy CBS-QB3 method, when feasible. Relative pK(a) values were estimated using the PCM model for aqueous solvation. The terminal oxygen is the most basic site, followed closely by the internal oxygen. Protonation of these sites does not lead to decomposition. However, protonation of the weakly basic amino nitrogen leads to very rapid decomposition and NO generation. PMID- 14753827 TI - X-ray absorption spectroscopic studies of chromium(V/IV/III)- 2-ethyl-2 hydroxybutanoato(2-/1-) complexes. AB - Structures of the complexes [Cr(V)O(ehba)(2)](-), [Cr(IV)O(ehbaH)(2)](0), and [Cr(III)(ehbaH)(2)(OH(2))(2)](+) (ehbaH(2) = 2-ethyl-2-hydroxybutanoic acid) in frozen aqueous solutions (10 K, [Cr] = 10 mM, 1.0 M ehbaH(2)/ehbaH, pH 3.5) have been determined by single- and multiple-scattering fitting of X-ray absorption fine structure (XAFS) data. An optimal set of fitting parameters has been determined from the XAFS calculations for a compound with known crystal structure, Na[Cr(V)O(ehba)(2)] (solid, 10 K). The structure of the Cr(V) complex [Cr(V)O(ehba)(2)](-) does not change in solution in the presence of excess ligand. Contrary to the earlier suggestions made from the kinetic data (Ghosh, M. C.; Gould, E. S. J. Chem. Soc., Chem. Commun. 1992, 195-196), the structure of the Cr(IV) complex (generated by the Cr(VI) + As(III) + ehbaH(2) reaction) is close to that of the Cr(V) complex (five-coordinate, distorted trigonal bipyramidal) and different from that of the Cr(III) complex (six-coordinate, octahedral). For both Cr(V) and Cr(IV) complexes, some disorder in the position of the oxo group is observed, which is consistent with but not definitive for the presence of geometric isomers. The structure of the Cr(IV) complex differs from that of Cr(V) by protonation of alcoholato groups of the ligands, which leads to significant elongation of the corresponding Cr-O bonds (2.0 vs 1.8 A). This is reflected in the different chemical properties reported previously for the Cr(IV) and Cr(V) complexes, including their reactivities toward DNA and other biomolecules in relation to Cr-induced carcinogenicity. PMID- 14753828 TI - Stepwise synthesis of [Ru(trpy)(L)(X)](n+) (trpy = 2,2':6',2' '-terpyridine; L = 2,2'-dipyridylamine; X = Cl-, H2O, NO2-, NO+, O2-). Crystal structure, spectral, electron-transfer, and photophysical aspects. AB - Ruthenium-terpyridine complexes incorporating a 2,2'-dipyridylamine ancillary ligand [Ru(II)(trpy)(L)(X)](ClO(4))(n) [trpy = 2,2':6',2' '-terpyridine; L = 2,2' dipyridylamine; and X = Cl(-), n = 1 (1); X = H(2)O, n = 2 (2); X = NO(2)(-), n = 1 (3); X = NO(+), n = 3 (4)] were synthesized in a stepwise manner starting from Ru(III)(trpy)(Cl)(3). The single-crystal X-ray structures of all of the four members (1-4) were determined. The Ru(III)/Ru(II) couple of 1 and 3 appeared at 0.64 and 0.88 V versus the saturated calomel electrode in acetonitrile. The aqua complex 2 exhibited a metal-based couple at 0.48 V in water, and the potential increased linearly with the decrease in pH. The electron-proton content of the redox process over the pH range of 6.8-1.0 was calculated to be a 2e(-)/1H(+) process. However, the chemical oxidation of 2 by an aq Ce(IV) solution in 1 N H(2)SO(4) led to the direct formation of corresponding oxo species [Ru(IV)(trpy)(L)(O)](2+) via the concerted 2e(-)/2H(+) oxidation process. The two successive reductions of the coordinated nitrosyl function of 4 appeared at +0.34 and -0.34 V corresponding to Ru(II)-NO(+) --> Ru(II)-NO* and Ru(II)-NO* --> Ru(II)-NO(-), respectively. The one-electron-reduced Ru(II)-NO* species exhibited a free-radical electron paramagnetic resonance signal at g = 1.990 with nitrogen hyperfine structures at 77 K. The NO stretching frequency of 4 (1945 cm(-1)) was shifted to 1830 cm(-1) in the case of [Ru(II)(trpy)(L)(NO*)](2+). In aqueous solution, the nitrosyl complex 4 slowly transformed to the nitro derivative 3 with the pseudo-first-order rate constant of k(298)/s(-1) = 1.7 x 10(-4). The chloro complex 1 exhibited a dual luminescence at 650 and 715 nm with excited state lifetimes of 6 and 1 micros, respectively. PMID- 14753829 TI - Auto-ionization in lutetium iodide complexes: effect of the Iioic radius on lanthanide-iodide binding. AB - Reaction of lutetium metal with 1.5 equiv of elemental iodine in 2-propanol leads to the isolation of LuI(3)(HO(i)Pr)(4) (1). An X-ray crystal structure reveals an ionic structure with well-separated [LuI(2)(HO(i)Pr)(4)] cations and [I] anions. Dissolution of 1 in pyridine generates the unusual alkoxide species [LuI(O(i)Pr)(py)(5)][I] (2) with the elimination of HI. An X-ray crystal structure of 2 confirmed the ionic nature of the compound, with the cationic portion of the complex exhibiting a seven-coordinated lutetium center with trans disposed iodo and alkoxide ligands and five pyridine molecules equally displaced within the equatorial plane. Exposure of 2 to iodotrimethylsilane yields the expected triiodide species [LuI(2)(py)(5)][I] (3), which may also be prepared by refluxing commercially available LuI(3) in THF, followed by crystallization from a THF/pyridine mixture. The solid-state structure of 3 is similar to that of 2, with the alkoxide ligand having been replaced by an iodide. The formation of ionic structures 1-3 as opposed to the higher-coordinated neutral species may be traced to the small lutetium center and the presence of relatively strong Lewis bases within the coordination sphere of the metal. PMID- 14753830 TI - Structural, spectroscopic, and electrochemical studies of the complexes [Ni2(mu CNR)(CNR)2(mu-dppm)2](n+) (n = 0, 1, 2): unusual examples of nickel(0)-nickel(I) and nickel(0)-nickel(II) mixed valency. AB - Reaction of Ni(COD)(2) (COD = cyclooctadiene) with dppm (dppm = bis(diphenylphosphino) methane) followed by addition of alkyl or aryl isocyanides yields the class of nickel(0) dimers Ni(2)(mu-CNR)(CNR)(2)(mu-dppm)(2) (R = CH(3) (1), n-C(4)H(9) (2), CH(2)C(6)H(5) (3), i-C(3)H(7) (4), C(6)H(11) (5), t-C(4)H(9) (6), p-IC(6)H(4) (7), 2,6-(CH(3))(2)C(6)H(3) (8)). The cyclic voltammograms of the dimers exhibit two sequential single electron oxidations to the +1 and +2 forms. Specular reflectance infrared spectroelectrochemical (IRSEC) measurements demonstrate reversible interconversions between the neutral Ni(0) dimers and their +1 and +2 forms. Bulk samples of the +2 forms are prepared by chemical oxidation using [FeCp(2)][PF(6)], while the +1 forms are prepared by the comproportionation of neutral and +2 forms. The neutral complexes 6 and 8 were characterized by X-ray diffraction as symmetric, locally tetrahedral binuclear Ni(0) complexes. The +2 forms of these complexes, 6(2+) and 8(2+), have asymmetric structures with one locally square planar and one locally tetrahedral metal center, evidence for a Ni(II)-Ni(0) mixed valence state. The X-ray structural characterization of 6(+) is symmetrical and qualitatively similar to that of the neutral complex 6. The +1 forms all exhibit intense near IR electronic absorptions that are assigned as intervalence charge transfer (IVCT) bands. On the basis of structural, spectroscopic, and electrochemical data, the +1 forms of the complexes, 1(+)-8(+), are assigned as Robin-Day class III, fully delocalized Ni(+0.5)-Ni(+0.5) mixed valence complexes. PMID- 14753831 TI - Syntheses, structure, and selected physical properties of CsLnMnSe3 (Ln = Sm, Gd, Tb, Dy, Ho, Er, Tm, Yb, Y) and AYbZnQ3 (A = Rb, Cs; Q = S, Se, Te). AB - CsLnMnSe(3) (Ln = Sm, Gd, Tb, Dy, Ho, Er, Tm, Yb, Y) and AYbZnQ(3) (A = Rb, Cs; Q = S, Se, Te) have been synthesized from solid-state reactions at temperatures in excess 1173 K. These isostructural materials crystallize in the layered KZrCuS(3) structure type in the orthorhombic space group Cmcm. The structure is composed of LnQ(6) octahedra and MQ(4) tetrahedra that share edges to form [LnMQ(3)] layers. These layers stack perpendicular to [010] and are separated by layers of face- and edge-sharing AQ(8) bicapped trigonal prisms. There are no Q-Q bonds in the structure of the ALnMQ(3) compounds so the formal oxidation states of A/Ln/M/Q are 1+/3+/2+/2-. The CsLnMnSe(3) materials, with the exception of CsYbMnSe(3), are Curie-Weiss paramagnets between 5 and 300 K. The magnetic susceptibility data for CsYbZnS(3), RbYbZnSe(3), and CsYbMSe(3) (M = Mn, Zn) show a weak cusp at approximately 10 K and pronounced differences between field-cooled and zero-field cooled data. However, CsYbZnSe(3) is not an antiferromagnet because a neutron diffraction study indicates that CsYbZnSe(3) shows neither long-range magnetic ordering nor a phase change between 4 and 295 K. Nor is the compound a spin glass because the transition at 10 K does not depend on ac frequency. The optical band gaps of the (010) and (001) crystal faces for CsYbMnSe(3) are 1.60 and 1.59 eV, respectively; the optical band of the (010) crystal faces for CsYbZnS(3) and RbYbZnSe(3) are 2.61 and 2.07 eV, respectively. PMID- 14753832 TI - Linear hybrid aminoborane/phosphinoborane chains: synthesis, proton-hydride interactions, and thermolysis behavior. AB - The reaction of the lithiated phosphine-borane adducts Li[PPhR.BH(3)] or Li[CH(2) PR(2).BH(3)] with Me(2)NH.BH(2)Cl afforded the hybrid linear species Me(2)NH BH(2)-PPhR-BH(3) (1, R = Ph; 2, R = H) or Me(2)NH-BH(2)-CH(2)-PR(2)-BH(3) (3, R = Ph; 4, R = Me). Single-crystal X-ray diffraction studies on 1 and 3, the first for linear hybrid aminoborane/phosphinoborane adducts, confirmed the expected four-coordinate N-B-P-B and N-B-C-P-B frameworks. In addition, interactions between the protic N-H and hydridic B-H hydrogen atoms resulted in short intermolecular H...H contacts for 1, whereas 3 was found to possess an exceptionally short intramolecular H...H distance of 1.95 A. Solution and solid state infrared studies on 3 and 4 also suggest that these dihydrogen interactions were maintained even in dilute solution. Hydrogen bond strengths in the range of 7.9 to 10.9 kJ mol(-1) indicate the presence of a relatively weak interaction. The thermal and catalytic dehydrocoupling reactivities of 1-4 were also investigated. Chain cleavage reactions were observed for 1 and 2 upon thermolysis at 130 degrees C to afford species such as Me(2)NH.BH(3), [Me(2)N-BH(2)](2), PhPRH.BH(3) (R = Ph, H), PhPRH (R = Ph, H), Ph(2)PH-BH(2)-PPh(2)-BH(3), and also the low molecular weight polyphosphinoborane [PhPH-BH(2)](n) (M(w) approximately 5000). Similar products were observed for the attempted catalytic dehydrocoupling reactions but under milder reaction conditions (50 degrees C). Thermolysis of 3 at 130 degrees C yielded the six-membered ring [BH(2)-CH(2)-PPh(2)](2) (5), which presumably results from the dissociation of Me(2)NH.BH(3) from 3. Thermolysis of 4 at 90 degrees C afforded Me(2)NH.BH(3) and Me(3)P.BH(3), in addition to a product tentatively assigned as [BH(2)-CH(2)-PMe(2)](2) (6). PMID- 14753833 TI - Dinuclear asymmetric ruthenium complexes with 5-cyano-1,10-phenanthroline as a bridging ligand. AB - New dinuclear asymmetric ruthenium complexes of the type [(bpy)(2)Ru(5 CNphen)Ru(NH(3))(5)](4+/5+) (bpy = 2,2'-bipyridine; 5-CNphen = 5-cyano-1,10 phenanthroline) have been synthesized and characterized by spectroscopic, electrochemical, and photophysical techniques. The structure of the cation [(bpy)(2)Ru(5-CNphen)Ru(NH(3))(5)](4+) has been determined by X-ray diffraction. The mononuclear precursor [Ru(bpy)(2)(5-CNphen)](2+) has also been prepared and studied; while its properties as a photosensitizer are similar to those of [Ru(bpy)(3)](2+), its luminescence at room temperature is quenched by a factor of 5 in the mixed-valent species [(bpy)(2)Ru(II)(5-CNphen)Ru(III)(NH(3))(5)](5+), pointing to the occurrence of intramolecular electron-transfer processes that follow light excitation. From spectral data for the metal-to-metal charge transfer transition Ru(II) --> Ru(III) in this latter complex, a slight electronic interaction (H(AB) = 190 cm(-1)) is disclosed between both metallic centers through the bridging 5-CNphen. PMID- 14753834 TI - Divalent molybdenum complexes of the dipyrrolide ligand system. Isolation of a Mo2 unit with a 45 degree twist angle. AB - The preparation of divalent Mo complexes of dipyrrolide dianions was carried out by reacting Mo(2)(acetate)(4) with the dipotassium salts of Ph(2)C(2 C(4)H(3)NH)(2) and 2-[1,1-bis(1H-pyrrol-2-yl)ethyl]pyridine. The two reactions respectively afforded the diamagnetic [[Ph(2)C(C(4)H(3)N)(2)](2)Mo(2)(OAc)(2)[K(THF)(3)][K(THF)]].THF (1) and [[(2 C(5)H(4) N)(CH(3))C(2-C(4)H(3)N)(2)]Mo(OAc)[K(THF)]](2).THF (2). Both compounds retained two acetate units in the dimetallic structure. Conversely, the reaction of Me(8)Mo(2)Li(4)(THF)(4) with Et(2)C(2-C(4)H(3)NH)(2) afforded the paramagnetic dimer [[Et(2)C(C(4)H(3)N)(2)](4)Mo(2)Li(2)][Li(THF)(4)](2).0.5THF (3). The paramagnetism is most likely caused by the 45 degree rotation of the two Mo(dipyrrolide) units with respect to each other and which, in turn, is caused by the presence of two lithium cations in the molecular structure. PMID- 14753835 TI - Density functional theory study of the d(10) series (H3P)3M(eta 1-SO2) and (MenPh(3-n)P)3M(eta 1-SO2) (M = Ni, Pd, Pt; n = 0-3): SO2 pyramidality and M-S bond dissociation energies. AB - Quantum mechanical density functional theory (DFT) and coupled DFT/molecular mechanics (QMMM) studies of the compounds (H(3)P)(3)M(eta(1)-SO(2)) and (Me(n)Ph(3-n)P)(3)M(eta(1)-SO(2)) (M = Ni, Pd, Pt; n = 0-3) model the experimental data well, particularly the planar/pyramidal geometries at sulfur. Bond dissociation energy (BDE) calculations confirm that Pd/Pt systems with pyramidal SO(2) ligands exhibit M-S BDEs smaller by 30-50% than Ni systems with planar SO(2). However, scans of the potential energy surfaces show that flexing the planar/pyramidal torsion angle within ranges of 20-30 degrees requires little energy. Bond energy decomposition calculations indicate that the electrostatic Delta E(elstat) term determines the BDE for Pd/Pt molecules where the sulfur is pyramidal, whereas all three terms matter when the sulfur is planar, as for Ni compounds. However, this accounts only for a fraction of the BDE differences; orbital energy matching accounts for the balance. PMID- 14753836 TI - Syntheses, structures, and physical properties of nickel bis(dithiolene) complexes containing tetrathiafulvalene (TTF) units. AB - To contribute to the development of single-component molecular metals, several nickel complexes with cyclohexeno-condensed or ethylenedioxy-substituted TTF (tetrathiafulvalene) dithiolate ligands, (R(4)N)(n)[Ni(chdt)(2)] [R = Me, n = 2 (1); R = (n)Bu, n = 1 (2); n = 0 (3)] and (R(4)N)(n)[Ni(eodt)(2)] [R = Me, n = 2 (4); R = (n)Bu, n = 1 (5); n = 0 (6)], were prepared. X-ray structures were determined on the monoanionic species 2 and 5. The tetra-n-butylammonium complex of the monoanionic [Ni(chdt)(2)] (2) with a 1:1 composition revealed that its magnetic susceptibility gave a good agreement with the Bonner-Fisher model (J/k(B) = -28 K), which was derived from the one-dimensional chains of anions with a regular interval. On the other hand, the magnetic susceptibility of the tetra-n-butylammonium complex of the monoanionic [Ni(eodt)(2)] (5) showed the Curie-Weiss behavior (C = 0.376 K.emu.mol(-1) and Theta = -4.6 K). Both of the monoanionic species 2 and 5 indicate that they belong to the S = 1/2 magnetic system and have relatively large and anisotropic g-values, suggesting the contribution of the nickel 3d orbital. Electrical resistivity measurements were performed on the compressed pellets of the neutral species 3 and 6. Fairly large conductivities were obtained (sigma(rt) = 1-10 S.cm(-1)). In addition, despite the measurements on the compressed pellets of powder samples, the neutral species 6 showed metallic behavior down to ca. 120 K and retained high conductivity even at 0.6 K [sigma(0.6 K)/sigma(rt) approximately 1/30], suggesting the crystal to be essentially metallic down to very low temperature. The electrical behavior and Pauli paramagnetism of 6 indicate the system to be a new single-component metal. PMID- 14753837 TI - Two-, three-, and four-coordinate Ag(I) coordination polymers formed by the novel phosphinite PPh2(3-OCH2C5H4N). AB - The novel phosphinite PPh(2)(3-OCH(2)C(5)H(4)N) (1) has been synthesized, and its coordination properties to Ag(I) have been studied. When reacted in a 1:1 ratio with Ag(I), coordination polymers with different coordination numbers about the Ag are found depending on the anion. For PPh(2)(3-OCH(2)C(5)H(4)N)AgBF(4) (2), a two-coordinate Ag is observed with a P-Ag-N angle of 167 degrees. Mixed three and four coordination about Ag is observed for PPh(2)(3-OCH(2)C(5)H(4)N)AgOTf (3), and for the trifluoroacetate derivative, PPh(2)(3-OCH(2)C(5)H(4)N)Agtfa (4), only a four-coordinate Ag is produced. X-ray crystal-structure determinations for compounds 2-4 have been carried out. The X-ray structures show a wide range of Ag Ag distances in the polymers, which are dependent on the conformation of the bridging ligand. PMID- 14753838 TI - Bacterial iron transport: coordination properties of azotobactin, the highly fluorescent siderophore of Azotobacter vinelandii. AB - Azotobacter vinelandii, a nitrogen-fixing soil bacterium, secretes in iron deficiency azotobactin delta, a highly fluorescent pyoverdin-like chromopeptidic hexadentate siderophore. The chromophore, derived from 2,3-diamino-6,7 dihydroxyquinoline, is bound to a peptide chain of 10 amino acids: (L)-Asp-(D) Ser-(L)-Hse-Gly-(D)-beta-threo-HOAsp-(L)-Ser-(D)-Cit-(L)-Hse-(L)-Hse lactone-(D) N(delta)-Acetyl, N(delta)-HOOrn. Azotobactin delta has three different iron(III) binding sites which are one hydroxamate group at the C-terminal end of the peptidic chain (N(delta)-Acetyl, N(delta)-HOOrn), one alpha-hydroxycarboxylic function in the middle of the chain (beta-threo-hydroxyaspartic acid), and one catechol group on the chromophore. The coordination properties of its iron(III) and iron(II) complexes were measured by spectrophotometry, potentiometry, and voltammetry after the determination of the acid-base functions of the uncomplexed free siderophore. Strongly negatively charged ferric species were observed at neutral p[H]'s corresponding to a predominant absolute configuration Lambda of the ferric complex in solution as deduced from CD measurements. The presence of an alpha-hydroxycarboxylic chelating group does not decrease the stability of the iron(III) complex when compared to the main trishydroxamate siderophores or to pyoverdins. The value of the redox potential of ferric azotobactin is highly consistent with a reductive step by physiological reductants for the iron release. Formation and dissociation kinetics of the azotobactin delta ferric complex point out that both ends of this long siderophore chain get coordinated to Fe(III) before the middle. The most striking result provided by fluorescence measurements is the lasting quenching of the fluorophore in the course of the protonation of the ferric azotobactin delta complex. Despite the release of the hydroxyacid and of the catechol, the fluorescence remains indeed quenched, when iron(III) is bound only to the hydroxamic acid, suggesting a folded conformation at this stage, around the metal ion, in contrast to the unfolded species observed for other siderophores such as ferrioxamine or pyoverdin PaA. PMID- 14753839 TI - Cation and pressure effects on the electrochemistry of 12-tungstocobaltate and 12 tungstophosphate ions in acidic aqueous solution. AB - The effects of supporting electrolytes and of pressure on the electrode reactions of the aqueous CoW(12)O(40)(5-/6-) couple at 25 degrees C are reported, together with limited data on PW(12)O(40)(3-)/4-) and PW(12)O(40)(4-/5-). The half-wave potentials E(1/2) for the CoW(12) couple become moderately more positive with increasing electrolyte concentration and cationic charge, and also in the sequences Li(+) approximately Na(+) < NH(4)(+) < or = H(+) < K(+) < Rb(+) < Cs(+) and Na(+) < Mg(2+) < Ca(2+) < Eu(3+). The mean diffusion coefficients for CoW(12) with the 1:1 electrolytes are independent of electrolyte concentration and rise only slightly from Li(+) to Cs(+), averaging (2.4 +/- 0.3) x 10(-6) cm(2) s(-1). Neither the volumes of activation for diffusion Delta V(diff)(++) (average -0.9 +/- 1.1 cm(3) mol(-1)) nor the electrochemical cell reaction volumes Delta V(Ag/AgCl) (average -22 +/- 2 cm(3) mol(-1)) for the CoW(12) couple show significant dependence on electrolyte identity or concentration. For the PW(12)(3 /4-) and PW(12)(4-/5-) couples, Delta V(Ag/AgCl) = -14 and -26 cm(3) mol(-1), respectively, suggesting a dependence on Delta(z(2)) (z = ionic charge number) as predicted by the Born-Drude-Nernst theory of electrostriction of solvent, but comparison with Delta V(Ag/AgCl) for CoW(12) and other anion-anion couples shows that the Born-Drude-Nernst approach fails in this context. For aqueous electrode reactions of CoW(12), as for other anionic couples such as cyanometalates, the standard rate constants k(el) show specific cation catalysis (Na(+) < K(+) < Rb(+) < Cs(+)), and Delta V(el++) is invariably positive, in the presence of supporting electrolytes. For the heavier group 1 cations, Delta V(el++) is particularly large (10-15 cm(3) mol(-1)), consistent with a partial dehydration of the cation to facilitate catalysis of the electron-transfer process. The positive values of Delta V(el++) for the CoW(12) couple cannot be attributed to rate control by solvent dynamics, which would lead to Delta V(el++) < or = Delta V(diff++), i.e., to negative or zero Delta V(el++) values. These results stand in sharp contrast to those for aqueous cationic couples, for which k(el) shows relatively little influence of the nature of the counterion and Delta V(el++) is always negative. PMID- 14753840 TI - Synthesis and characterization of binucleating bis(amidinate) ligands and their dialuminum complexes. AB - Two general routes to binucleating bis(amidinate) ligands based on dibenzofuran and 9,9-dimethylxanthene backbones are reported. The free-base form of one of the ligands, (Ph,Mes)L(DBF)H(2), forms a 1:1 adduct with acetone. Single-crystal X ray diffraction of this adduct reveals bidentate H-bonding of the bis(amidine) to the ketone oxygen. Bond lengths suggest that the individual H-bonds are relatively weak, yet IR spectroscopy shows a significant -26 cm(-1) shift for the carbonyl stretch relative to free acetone. Additionally, the new dialuminum complexes (i)(Pr)L(DBF)Al(2)Me(4) (3), (i)(Pr)L(Xan)Al(2)Me(4) (4), (t)(Bu,Et)L(DBF)Al(2)Me(4) (5), and (t)(Bu,Et)L(Xan)Al(2)Me(4) (6) are prepared by reaction of Al(2)Me(6) with the bis(amidines) in toluene solution. (1)H NMR spectroscopic studies indicate that 3 and 4 interact weakly with certain Lewis bases (DMSO, DMF, pyridine) to effect the exchange of the Al-bound Me groups. Other bases, such as THF and TMEDA, fail to interact. Solid-state structures for 3 and 4 are reported. PMID- 14753841 TI - Synthesis, structure, and magnetochemical analysis of selected first-row transition-metal anilino- and anisolesquarate compounds. AB - The isomorphous polymeric complexes [M(mu-C(6)H(5)NHC(4)O(3))(2)(CH(3)OH)(2)](n) [M = Mn (1), Co (2), Cu (4), Zn (5)] are produced by reacting the anilinosquarate anion with the appropriate metal nitrates in a methanolic solution. Each of these complexes contains the central metal atom in a slightly distorted octahedral environment, with the coordination polyhedron consisting of four mu-1,2-bridging anilinosquarate ligands and two trans-oriented methanols. The polymer chains propagate to form a two-dimensional net of metal centers, with the conformation of the component sheets in the net being controlled by intramolecular N-H...O and O-H...O hydrogen bonds. Under reaction conditions similar to those used in the synthesis of the polymers 1, 2, 4, and 5, the nickel(II) monomer [Ni(C(6)H(5)NHC(4)O(3))(2)(H(2)O)(4)].2H(2)O (3) is produced in which each nickel center is attached to two cis-coordinated anilinosquarate and four aqua ligands in a distorted octahedral arrangement. The ligand conformation in 3 is stabilized by both intra- and intermolecular hydrogen bonding, which results in the formation of a sheet polymer having distinct hydrophobic and hydrophilic surfaces. Magnetochemical analysis of 1 and 4 reveals normal paramagnetic behavior for 1 and a very weak ferromagnetic interaction in 4; the absence of significant magnetic interactions is attributed to the distortion of the C(4) cycle of the anilinosquarate ligand (lower than C(2)(v) symmetry) in these complexes. Reaction of anisolesquarate with M(NO(3))(2).xH(2)O in acetonitrile produced the set of isomorphous salts [M(H(2)O)(6)][CH(3)OC(6)H(5)C(4)O(3)](2) [M = Mn (6), Co (7), Ni (8), Zn (9)]. The anisolesquarate anions in 6-9 are hydrogen bonded to the [M(H(2)O)(6)](2+) ions to form polymer chains, which are further linked by hydrogen bonds to form complex sheets. Complexation of the anisolesquarate ligand was not observed even when other solvents and reaction conditions were employed. PMID- 14753842 TI - Effect of equatorial ligands of dirhodium(II,II) complexes on the efficiency and mechanism of transcription inhibition in vitro. AB - The nature of the equatorial ligands spanning the dirhodium core was shown to affect the ability and mechanism of various lantern-type complexes to inhibit transcription in vitro. The inhibition of transcription by Rh(2)(mu O(2)CCF(3))(4), Rh(2)(mu-HNCOCF(3))(4), and [Rh(2)(mu O(2)CCH(3))(2)(CH(3)CN)(6)](2+) appears to proceed predominantly via binding of the complexes to T7-RNA polymerase (T7-RNAP) and is dependent on the concentration of enzyme and Mg(2+) ions in solution. The concentrations of the aforementioned complexes required to inhibit 50% of the transcription, C(inh)(50), are similar to that measured for activated cisplatin, whereas a significantly higher concentration of Rh(2)(mu-HNCOCH(3))(4) is required to effect similar inhibition; the inhibition induced by Rh(2)(mu-HNCOCH(3))(4) does not involve binding to T7-RNAP. The spectral changes observed for each complex upon addition of enzyme are consistent with Rh(2)(mu-O(2)CCF(3))(4), Rh(2)(mu HNCOCF(3))(4), and [Rh(2)(mu-O(2)CCH(3))(2)(CH(3)CN)(6)](2+) binding to the enzyme and may involve partial displacement of the equatorial (eq) groups by the Lewis basic sites of T7-RNAP. In contrast, addition of enzyme to solutions of Rh(2)(mu-HNCOCH(3))(4) does not result in significant spectral changes, a finding consistent with lack of enzyme dependence in the transcription inhibition. These differences in reactivity and transcription inhibition mechanism among complexes with different bridging ligands are explained by variations of the Lewis acidity of the axial (ax) sites in the series of complexes Rh(2)(mu-O(2)CCF(3))(4), Rh(2)(mu-HNCOCF(3))(4), and Rh(2)(mu-HNCOCH(3))(4). The Lewis acidity of the ax sites is expected to affect the initial interaction of the complexes with the biomolecules, followed by their rearrangement to eq positions if the bridging ligands are labile. PMID- 14753843 TI - Molecular structures of alumina nanoballs and nanotubes: a theoretical study. AB - Molecular structures of alumina nanoballs and nanotubes have been determined. Tetrahedral, octahedral, and icosahedral alumina nanostructures were derived from Platonic solids and Archimedean polyhedra and were optimized by quantum chemical methods. I(h)-symmetric balls, resembling their isovalence electronic analogues, fullerenes, are preferred. The nanoballs consist of adjacent Al(5)O(5) and Al(6)O(6) rings, similar to C(5)- and C(6)-rings of fullerenes. The structural characteristics of alumina nanoballs are dominated by pi-electron donation from oxygen to aluminum. Alumina nanotubes can be derived from icosahedral nanoballs. The tubes alternate between D(5d)- and D(5h)-symmetries and are capped by halves of the icosahedral balls. PMID- 14753844 TI - Effect of amine ligand bulk on the interaction of methionine with platinum(II) diamine complexes. AB - Molecular mechanics and dynamics calculations have been used in conjunction with experimental data to study the effects of amine ligand bulk on the formation of both guanine and methionine complexes with platinum diamine compounds. The AMBER force field has been supplemented with previous modifications [Yao; et al. Inorg. Chem. 1994, 33, 6061-6077. Cerasino; et al. Inorg. Chem. 1997, 36, 6070-6079] and has been further modified to include parameters for platinum bound to the sulfur atom of methionine. Molecular mechanics calculations with this modified AMBER force field have suggested that a platinum complex with two sulfur-bound methionine ligands and a bulky diamine ligand (N,N,N',N' tetramethylethylenediamine, Me(4)en) would have severe interligand clashes; such interligand clashes are less pronounced in bis(9-ethylguanine) complexes. Consistent with these observations, NMR studies with [Pt(Me(4)en)(D(2)O)(2)](2+) have indicated that guanine 5'-monophosphate reacts in a 2:1 guanine:platinum ratio while both methionine and N-acetylmethionine react with only a 1:1 stoichiometry. Methionine forms a chelate via the sulfur and nitrogen atoms whereas N-acetylmethionine forms a chelate via the sulfur and oxygen atoms. The oxygen of the latter chelate can be displaced by the addition of guanosine 5' monophosphate, although complete displacement of the N-acetylmethionine was not observed. PMID- 14753845 TI - Luminescent heterotrinuclear complexes with Pt(diimine)(dithiolate) and metal diphosphine as components. AB - Reactions of Pt(diimine)(tdt) (tdt =3,4-toluenedithiolate) with [M(2)(dppm)(2)(MeCN)(2)](2+) (M = Cu(I) or Ag(I), dppm = bis(diphenylphosphino)methane) gave heterotrinuclear complexes [PtCu(2)(tdt)(mu SH)(dppm)(3)](ClO(4)) (1) and [PtCu(2)(diimine)(2)(tdt)(dppm)(2)](ClO(4))(2) (diimine = 2,2'-bpyridine (bpy) 2; 4,4'-dimethyl-2,2'-bipyridine (dmbpy) 3; phenanthroline (phen) 4, 5-bromophenanthroline (Brphen) 5) for M = Cu(I), but [PtAg(2)(tdt)(mu-SH)(dppm)(3)](SbF(6)) (6) and [PtAg(2)(diimine)(tdt)(dppm)(2)](SbF(6))(2) (diimine = bpy 7; dmbpy 8; phen 9; Brphen 10) for M = Ag(I). While the complexes [PtAg(2)(diimine)(tdt)(dppm)(2)](SbF(6))(2) (7-10) result from linkage of Pt(diimine)(tdt) and [M(2)(dppm)(2)(MeCN)(2)](2+) by tdt sulfur donors, formation of [PtCu(2)(diimine)(2)(tdt)(dppm)(2)](ClO(4))(2) (2-5) is related to rupture of metal-ligand bonds in the metal components and recombination between the ligands and the metal atoms by self-assembly. The formation of 1 and 6 is involved not only in dissociation and recombination of the metal components, but also in disruption of C-S bonds in the dithiolate (tdt). The dithiolate tdt adopts a chelating and bridging coordination mode in anti conformation for [PtCu(2)(diimine)(2)(tdt)(dppm)(2)](ClO(4))(2) (2-5), whereas there is the syn conformation for other complexes. Compounds 1 and 6 represent sparse examples of mu-SH-bridged heterotrinuclear Pt(II)M(I)(2) complexes, in which Pt(II)-M(I) centers are bridged by dppm and sulfur donors of tdt, whereas M(I)-M(I) (M = Cu for 1; Ag for 6) centers are linked by dppm and the mu-SH donor. The (31)P NMR spectra show typical platinum satellites (J(Pt-P) = 1450-1570 Hz) for 1-6 and Ag P coupling for Pt(II)-Ag(I) (J(Ag-P) = 350-450 Hz) complexes 6-10. All of the complexes show intense emission in the solid state and in frozen glasses at 77 K. The complexes [PtAg(2)(diimine)(tdt)(dppm)(2)](SbF(6))(2) (7-10) also afford emission in fluid acetonitrile solutions at room temperature. Solid-state emission lifetimes at room temperature are in the microsecond range. It is revealed that emission energies of the trinuclear heterometallic complexes [PtAg(2)(diimine)(tdt)(dppm)(2)](SbF(6))(2) (7-10) exhibit a remarkable blue shift (0.10-0.35 eV) relative to those of the precursor compounds Pt(diimine)(tdt). The crystal structures of 1, 2, 4, 6, 8, and 9 were determined by X-ray crystallography. PMID- 14753847 TI - Identification and authentication of animal cell culture by polymerase chain reaction amplification and DNA sequencing. AB - Polymerase chain reaction (PCR) amplification and deoxyribonucleic acid (DNA) sequence analysis were used to identify the species origin of cell lines used in a cell culture facility where various cell lines of different species are routinely propagated. The aldolase gene family was selected for PCR amplification because the DNA sequences of this gene are highly conserved over a wide range of animals and humans. A total of 36 cell lines representing 13 different species were selected for this study. The DNA from each cell line was amplified, and PCR products were analyzed by agarose gel electrophoresis. The results showed unique profiles of amplified bands on agarose gels that allowed differentiation among non-closely related species. However, DNA amplification of closely related species, including rat and mouse or human and primate, resulted in similar and indistinguishable banding patterns that could be further differentiated by DNA sequence analysis. These results suggested that aldolase gene amplification coupled with DNA sequence analysis is a useful tool for identification of cell lines and has potential application for use in identification of interspecies cross-contamination. PMID- 14753848 TI - Action of major insecticide groups on insect cell lines of the beet armyworm, Spodoptera exigua, compared with larvicidal toxicity. AB - In a first series of experiments, the biological response of a continuous cell line of the beet armyworm, Spodoptera exigua, was tested with different groups of insecticides with different modes of actions: acetylcholinesterase inhibitors, acetycholine receptor agonists, inhibitors and uncouplers of oxidative phosphorylation, site I electron transport inhibitors, gamma-aminobutyric acid receptor inhibitors, chitin synthesis inhibitors, and juvenile hormone analogues. From the concentration response curves, 50% inhibition concentration (IC(50)) values were calculated. The most active compound in vitro was pyridaben with an IC(50) value of 0.0083 ppm. In a second series of experiments, the toxicity of these insecticide groups was determined on third-instar larvae of S. exigua, and lethal concentration with 50% kill (LC(50)) values were used in the evaluation of their in vivo biological activity. Toxicity bioassays showed that lufenuron was the most toxic (LC(50) = 0.098 ppm). To explain the discrepancies in biological responses in vitro with insect cells compared with in vivo conditions with whole third-instar larvae, the significance of different detoxifying enzyme systems was tested. P(450) monooxygenases, esterases, and glutathione S-transferases were measured in third-instar larvae and cells of S. exigua. Data are discussed in terms of the usefulness of insect cell cultures as tools in the screening for novel insecticide actions. PMID- 14753850 TI - Fortification of a protein-free cell culture medium with plant peptones improves cultivation and productivity of an interferon-gamma-producing CHO cell line. AB - A strong tendency is currently emerging to remove not only serum but also any product of animal origin from animal cell culture media during production of recombinant proteins. This should facilitate downstream processing and improve biosafety. One way consists in the fortification of protein-free nutritive media with plant protein hydrolysates. To investigate the effects of plant peptones on mammalian cell cultivation and productivity, CHO 320 cells, a clone of CHO K1 cells genetically modified to secrete human interferon-gamma (IFN-gamma), were first adapted to cultivation in suspension in a protein-free medium. Both cell growth and IFN-gamma secretion were found to be equivalent to those reached in serum-containing medium. Eight plant peptones, selected on the basis of their content in free amino acids and oligopeptides, as well as molecular weight distribution of oligopeptides, were tested for their ability to improve culture parameters. These were improved in the presence of three peptones, all having an important fraction of oligopeptides ranging from 1 to 10 kDa and a small proportion of peptides higher than 10 kDa. These peptones do not seem to add significantly to the nutritive potential to basal protein-free nutritive medium. Nevertheless, supplementation of an oligopeptide-enriched wheat peptone improved cell growth by up to 30% and IFN-gamma production by up to 60% in shake-flask experiments. These results suggest that the use of plant peptones with potential growth factor-like or antiapoptotic bioactivities could improve mammalian cell cultivation in protein-free media while increasing the product biosafety. PMID- 14753851 TI - Quantum Zeno effect for exponentially decaying systems. AB - The quantum Zeno effect--suppression of decay by frequent measurements--was believed to occur only when the response of the detector is so quick that the initial tiny deviation from the exponential decay law is detectable. However, we show that it can occur even for exactly exponentially decaying systems, for which this condition is never satisfied, by considering a realistic case where the detector has a finite energy band of detection. The conventional theories correspond to the limit of an infinite bandwidth. This implies that the Zeno effect occurs more widely than expected thus far. PMID- 14753849 TI - Expression of fibroblast growth factor receptor genes in human hepatoma-derived cell lines. AB - The fibroblast growth factor (FGF) function has been considered to contribute to various human tumors and malignant growth of neoplasm. Hepatocellular carcinoma (HCC) is a typical hypervascular tumor, and it is suggested that FGF may be involved in hepatocarcinogenesis. Therefore, the relationship between the progression of HCC and expression of FGFs and FGF receptors (FGFRs) was evaluated in this study. We investigated the expression of messenger ribonucleic acids (mRNAs) of FGFs and FGFRs by reverse transcriptase-polymerase chain reaction (RT PCR) analysis in eight human hepatoma-derived cell lines (Hep3B, HLE, HLF, HUH6, HUH7, KIM1, Li7, and PLC/PRF/5), one hepatoblastoma-derived cell line (HepG2), and human primary hepatocytes. In addition, effects of FGF-1, FGF-2, and FGF-7 on the growth of hepatoma-derived cell lines were studied in serum-free defined culture conditions. An RT-PCR analysis revealed that all cell lines except PLC/PRF/5 expressed all FGFR mRNAs: FGF-R1 (IIIc), -R2 (IIIb), -R2 (IIIc), -R3 (IIIb), -R3 (IIIc), and -R4 mRNAs. In contrast, human primary hepatocytes expressed FGF-R1 (IIIc), -R3 (IIIc), and -R4 mRNAs but not mRNAs of FGF-R2 (IIIb), -R2 (IIIc), and -R3 (IIIb). All cell lines except HUH6 and HUH7 expressed FGF-1 and FGF-2 mRNAs. Addition of exogenous FGF-1 or FGF-2 (or both) to culture stimulated cell proliferation in several cell lines, but FGF-7 exhibited no growth stimulation in all cells. Hepatoma cells may possess a proliferation mechanism regulated by an autocrine mechanism, a paracrine mechanism, or both, which are mediated by FGF-1/FGFR or FGF-2/FGFR (or both). In addition, a gain of FGF-R2 (IIIb), -R2 (IIIc), and -R3 (IIIb) may be associated with malignant transformation of liver tumor and may eventually serve as useful diagnostic and prognostic indicators. PMID- 14753852 TI - Quasi-one-dimensional Bose gases with a large scattering length. AB - Bose gases confined in highly elongated harmonic traps are investigated over a wide range of interaction strengths using quantum Monte Carlo techniques. We find that the properties of a Bose gas under tight transverse confinement are well reproduced by a 1D model Hamiltonian with contact interactions. We point out the existence of a unitary regime, where the properties of the quasi-1D Bose gas become independent of the actual value of the 3D scattering length a(3D). In this unitary regime, the energy of the system is well described by a hard-rod equation of state. We investigate the stability of quasi-1D Bose gases with positive and negative a(3D). PMID- 14753853 TI - Superfluid-superfluid phase transitions in a two-component Bose-Einstein condensate. AB - Depending on the Hamiltonian parameters, two-component bosons in an optical lattice can form at least three different superfluid phases in which both components participate in the superflow: a (strongly interacting) mixture of two miscible superfluids (2SF), a paired superfluid (PSF) vacuum, and (at a commensurate total filling factor) the super-counter-fluid (SCF) state. We study the universal properties of the 2SF-PSF and 2SF-SCF quantum phase transitions and show that (i) they can be mapped onto each other and (ii) their universality class is identical to the (d+1)-dimensional normal-superfluid transition in a single-component liquid. The finite-temperature 2SF-PSF(SCF) transitions and the topological properties of 2SF-PSF(SCF) interfaces are also discussed. PMID- 14753854 TI - Irregular dynamics in a one-dimensional Bose system. AB - We study many-body quantum dynamics of delta-interacting bosons confined in a one dimensional ring. Main attention is paid to the transition from the mean-field to the Tonks-Girardeau regime using an approach developed in the theory of interacting particles. We analyze, both analytically and numerically, how the Shannon entropy of the wave function and the momentum distribution depend on time for weak and strong interactions. We show that the transition from regular (quasiperiodic) to irregular ("chaotic") dynamics coincides with the onset of the Tonks-Girardeau regime. In the latter regime, the momentum distribution of the system reveals a statistical relaxation to a steady state distribution. The transition can be observed experimentally by studying the interference fringes obtained after releasing the trap and letting the boson system expand ballistically. PMID- 14753855 TI - Critical temperature of a trapped, weakly interacting Bose gas. AB - We report on measurements of the critical temperature of a harmonically trapped, weakly interacting Bose gas as a function of atom number. Our results exclude ideal-gas behavior by more than two standard deviations, and agree quantitatively with mean-field theory. At our level of sensitivity, we find no additional shift due to critical fluctuations. In the course of this measurement, the onset of hydrodynamic expansion in the thermal component has been observed. Our thermometry method takes this feature into account. PMID- 14753856 TI - Link invariants of electromagnetic fields. AB - The cross-helicity integral is known in fluid dynamics and plasma physics as a topological invariant which measures the mutual linkage of two divergence-free vector fields, e.g., magnetic fields, on a three-dimensional domain. Generalizing this concept, a new topological invariant is found which measures the mutual linkage of three closed two-forms, e.g., electromagnetic fields, on a four dimensional domain. The integral is shown to detect a separation of the cross helicity between two of the fields with the help of the third field. It can be related to the triple linking number known in knot theory. Furthermore, it is shown that the well-known three-dimensional cross helicity and the new four dimensional invariant are the first two examples of a series of topological invariants which are defined by n-1 field strengths F=dA on a simply connected n dimensional manifold M(n). PMID- 14753857 TI - Nonsymmetric entanglement of atomic ensembles. AB - The entanglement of multiatom quantum states is considered. In order to cancel noise due to inhomogeneous light-atom coupling, the concept of matched multiatom observables is proposed. As a means to eliminate an important form of decoherence this idea should be of broad relevance for quantum information processing with atomic ensembles. The general approach is illustrated on the example of rotation angle measurement, and it is shown that the multiatom states that were thought to be only weakly entangled can exhibit near-maximum entanglement. PMID- 14753858 TI - Quantum Creep and Quantum-Creep Transitions in 1D Sine-Gordon Chains. AB - Discrete sine-Gordon (SG) chains are studied with path-integral molecular dynamics. Chains commensurate with the substrate show the transition from pinning to quantum creep at bead masses slightly larger than in the continuous SG model. Within the creep regime, a field-driven transition from creep to complete depinning is identified. The effects of disorder on the chain's dynamics depend on the potential's roughness exponent H. For example, quantum fluctuations are generally too small to depin the chain if H=1/2, while an H=0 chain can be pinned or unpinned depending on the bead masses. Thermal fluctuations always depin the chain. PMID- 14753859 TI - Kac limit for finite-range spin glasses. AB - We consider a finite-range spin glass model in arbitrary dimension, where the strength of the two-body coupling decays to zero over some distance gamma(-1). We show that, under mild assumptions on the interaction potential, the infinite volume free energy of the system converges to that of the Sherrington-Kirkpatrick one, in the Kac limit gamma-->0. This could be a first step toward an expansion around mean-field theory, for spin glass systems. PMID- 14753860 TI - Reevaluation of the 22Na(p,gamma) reaction rate: implications for the detection of 22Na gamma rays from novae. AB - Understanding the processes which create and destroy 22Na is important for diagnosing classical nova outbursts. Conventional 22Na(p,gamma) studies are complicated by the need to employ radioactive targets. In contrast, we have formed the particle-unbound states of interest through the heavy-ion fusion reaction, 12C(12C,n)23Mg and used the Gammasphere array to investigate their radiative decay branches. Detailed spectroscopy was possible and the 22Na(p,gamma) reaction rate has been reevaluated. New hydrodynamical calculations incorporating the upper and lower limits on the new rate suggest a reduction in the yield of 22Na with respect to previous estimates, implying a reduction in the maximum detectability distance for 22Na gamma rays from novae. PMID- 14753861 TI - Decaying particles and the reionization history of the universe. AB - We investigate the possibility that the Universe is significantly reionized by the decay products of heavy particles. The ionization produced by decaying particles implies a high optical depth even if the maximum level of ionization ever produced is low (10(-2)). As a consequence, a high ionization fraction (x approximately equal to 0.5) at high redshifts (z approximately equal to 20) fails to fit the cosmic microwave background (CMB) spectra at l> or =30. Recent CMB data limit the primordial abundance of the decaying particles, favoring long decay times. Other significant sources of reionization are still needed at z approximately equal to 13. The decay process heats up the medium, bringing the expected y distortion to unobservable levels. PMID- 14753862 TI - Designing cyclic universe models. AB - The phenomenological constraints on the scalar field potential in cyclic models of the Universe are presented. We show that cyclic models require a comparable degree of tuning to that needed for inflationary models. The constraints are reduced to a set of simple design rules including "fast-roll" parameters analogous to the "slow-roll" parameters in inflation. PMID- 14753863 TI - Explosive dark matter annihilation. AB - In this Letter we study pair annihilation processes of dark matter (DM) in the Universe, in the case that the DM is an electroweak gauge nonsinglet. In the current Universe, in which the DM is highly nonrelativistic, the nonperturbative effect may enhance the DM annihilation cross sections, especially for that to two photons, by several orders of magnitude. We also discuss sensitivities in future searches for anomalous gamma rays from the galactic center, which originate from DM annihilation. PMID- 14753866 TI - Evidence for nonhadronic degrees of freedom in the transverse mass spectra of kaons from relativistic nucleus-nucleus collisions? AB - We investigate transverse hadron spectra from relativistic nucleus-nucleus collisions within two independent transport approaches that are based on quark, diquark, string, and hadronic degrees of freedom. Both transport models show their reliability for elementary pp as well as light-ion (C+C, Si+Si) reactions. However, for central Au+Au (Pb+Pb) collisions above approximately 5A GeV the measured K+/- transverse mass spectra have a larger inverse slope parameter than expected from the calculation. Thus, the pressure generated by hadronic interactions in the transport models above approximately 5A GeV is lower than observed in the experimental data. This finding shows that the additional pressure--as expected from lattice QCD calculations at finite quark chemical potential and temperature--is generated by strong partonic interactions in the early phase of central Au+Au (Pb+Pb) collisions. PMID- 14753865 TI - Semihard scattering unraveled from collective dynamics by two-pion azimuthal correlations in 158A GeV/c Pb+Au collisions. AB - Elliptic flow and two-particle azimuthal correlations of charged hadrons and high p(T) pions (p(T)>1 GeV/c) have been measured close to midrapidity in 158A GeV/c Pb+Au collisions by the CERES experiment. Elliptic flow (v(2)) rises linearly with p(T) to a value of about 10% at 2 GeV/c. Beyond p(T) approximately 1.5 GeV/c, the slope decreases considerably, possibly indicating a saturation of v(2) at high p(T). Two-pion azimuthal anisotropies for p(T)>1.2 GeV/c exceed the elliptic flow values by about 60% in midcentral collisions. These nonflow contributions are attributed to nearside and back-to-back jetlike correlations, the latter exhibiting centrality dependent broadening. PMID- 14753867 TI - Chiral degeneracy in triaxial 104Rh. AB - Chiral doublet bands based on the pi g(9/2) multiply sign in circle nu h(11/2) configuration that achieve degeneracy at spin I=17 in the odd-odd triaxial 104Rh nucleus have been observed. Experimental verification of the interpretation has been tested against specific fingerprints of chirality in the intrinsic system. PMID- 14753868 TI - High-order harmonic generation up to 250 eV from highly ionized argon. AB - We demonstrate the generation of very high-order harmonics, up to 250 eV, using argon gas. This extends by 100 eV the highest harmonics previously observed using Ar and exceeds the energies observed using any other medium besides helium. This advance is made possible by using a waveguide geometry to limit plasma-induced laser beam defocusing, making it possible to generate high harmonics from Ar ions. This work shows that high harmonic emission from ions can extend laser based coherent up-conversion into the soft x-ray region of the spectrum. PMID- 14753869 TI - Asymmetry in the strong-field ionization of Rydberg atoms by few-cycle pulses. AB - We present measurements of the electron ejection direction in the ionization of high (n=90) Rydberg states of rubidium subjected to few-cycle radio-frequency (RF) pulses. For weak pulses we find a strong asymmetry for even (cosine) pulses and no asymmetry for odd (sine) pulses. This asymmetry disappears for pulses longer than four RF cycles. For strong pulses, very large asymmetry is found for both sine and cosine pulses that persists up to eight RF cycles and is attributed to initial state depletion effects within a cycle. PMID- 14753864 TI - Observation of an exotic baryon with S=+1 in photoproduction from the proton. AB - The reaction gamma p-->pi(+)K(-)K(+)n was studied at Jefferson Laboratory using a tagged photon beam with an energy range of 3-5.47 GeV. A narrow baryon state with strangeness S=+1 and mass M=1555+/-10 MeV/c(2) was observed in the nK(+) invariant mass spectrum. The peak's width is consistent with the CLAS resolution (FWHM=26 MeV/c(2)), and its statistical significance is (7.8+/-1.0)sigma. A baryon with positive strangeness has exotic structure and cannot be described in the framework of the naive constituent quark model. The mass of the observed state is consistent with the mass predicted by the chiral soliton model for the Theta(+) baryon. In addition, the pK(+) invariant mass distribution was analyzed in the reaction gamma p-->K(-)K(+)p with high statistics in search of doubly charged exotic baryon states. No resonance structures were found in this spectrum. PMID- 14753870 TI - High-precision optical measurement of the 2S hyperfine interval in atomic hydrogen. AB - We have applied an optical method to the measurement of the 2S hyperfine interval in atomic hydrogen. The interval has been measured by means of two-photon spectroscopy of the 1S-2S transition on a hydrogen atomic beam shielded from external magnetic fields. The measured value of the 2S hyperfine interval is equal to 177 556 860(16) Hz and represents the most precise measurement of this interval to date. The theoretical evaluation of the specific combination of 1S and 2S hyperfine intervals D21 is in fair agreement (within 1.4 sigma) with the value for D21 deduced from our measurement. PMID- 14753871 TI - Production of ultracold, polar RbCs* molecules via photoassociation. AB - We have produced ultracold, polar RbCs* molecules via photoassociation in a laser cooled mixture of Rb and Cs atoms. Using a model of the RbCs* molecular interaction which reproduces the observed rovibrational structure, we infer decay rates in our experiments into deeply bound X(1)Sigma(+) ground-state RbCs vibrational levels as high as 5 x 10(5) s(-1) per level. Population in such deeply bound levels could be efficiently transferred to the vibrational ground state using a single stimulated Raman transition, opening the possibility to create large samples of stable, ultracold polar molecules. PMID- 14753872 TI - Nonhydrogenic effects in the deceleration of Rydberg atoms in inhomogeneous electric fields. AB - Argon atoms in Stark states at n approximately 25 have been decelerated and accelerated in inhomogeneous electric fields. The acceleration and deceleration behavior can be understood only by considering the adiabatic Landau-Zener dynamics that take place at the avoided crossings between the Stark states and the limited fluorescence lifetimes of approximately 10 micros. PMID- 14753873 TI - Control of atomic collisions by laser polarization. AB - Atomic collision pairs in a light field form a microscopic interferometer. The light acts as the beam splitter and controls at the same time the amplitudes and phases of the interfering waves. We demonstrate the complete tunability using linear and elliptic polarization. PMID- 14753874 TI - Quantum and classical coincidence imaging. AB - Coincidence, or ghost, imaging is a technique that uses two correlated optical fields to form an image of an object. In this work we identify aspects of coincidence imaging which can be performed with classically correlated light sources and aspects which require quantum entanglement. We find that entangled photons allow high-contrast, high-resolution imaging to be performed at any distance from the light source. We demonstrate this fact by forming ghost images in the near and far fields of an entangled photon source, noting that the product of the resolutions of these images is a factor of 3 better than that which is allowed by classical diffraction theory. PMID- 14753875 TI - Stable multicolor periodic-wave arrays. AB - We study the existence and stability of periodic-wave arrays propagating in uniform quadratic nonlinear media and discover that they become completely stable above a threshold light intensity. To the best of our knowledge, this is the first example in physics of completely stable periodic-wave patterns propagating in conservative uniform media supporting bright solitons. PMID- 14753876 TI - Time reversal and its application to tomography with diffracting sources. AB - An exact time-domain method is proposed to time reverse a transient scalar wave using only the field measured on an arbitrary closed surface enclosing the initial source. Under certain conditions, a time-reversed field can be approximated by retransmitting the measured signals in a reversed temporal order. Exact reconstruction for three-dimensional broadband diffraction tomography (a linearized inverse scattering problem) is proposed by time-reversing the measured field back to the time when each secondary source is excited. The algorithm is verified by a numerical simulation. Extension to the case using Green's function in a heterogeneous medium is discussed. PMID- 14753877 TI - Anisotropic weak localization of light. AB - We have observed angular anisotropy in weak localization of light from highly scattering, orientationally ordered, nematic liquid crystals. This demonstration of angular anisotropy in a multiple-scattering interference phenomenon was facilitated by a light scattering instrument with extraordinary angular resolution. The measured anisotropies were consistent with a simple model of coherent backscattering generalized for propagation-direction dependent mean free paths. PMID- 14753878 TI - Faster than Lyapunov decays of the classical Loschmidt echo. AB - We show that in the classical interaction picture the echo dynamics, namely, the composition of perturbed forward and unperturbed backward Hamiltonian evolution, can be treated as a time-dependent Hamiltonian system. For strongly chaotic (Anosov) systems we derive a cascade of exponential decays for the classical Loschmidt echo, starting with the leading Lyapunov exponent, followed by a sum of the two largest exponents, etc. In the loxodromic case a decay starts with the rate given as twice the largest Lyapunov exponent. For a class of perturbations of symplectic maps the echo dynamics exhibits a drift resulting in a superexponential decay of the Loschmidt echo. PMID- 14753879 TI - Bistability and "negative" viscosity for a suspension of insulating particles in an electric field. AB - It is shown that a suspension of insulating particles in a liquid with low conductivity possesses bistability and has a "negative" effective viscosity effect in the electric field due to internal rotations. By Brownian dynamics simulation it has been found that thermal fluctuations of the angular velocity of particles in this bistable system can have a large effect on the viscosity of the suspension. PMID- 14753880 TI - Formation of advanced tokamak plasmas without the use of an ohmic-heating solenoid. AB - A new operational scenario of advanced tokamak formation was demonstrated in the JT-60U tokamak. This was accomplished by electron cyclotron and lower hybrid waves, neutral beam injection, and the loop voltage supplied by the vertical field and shaping coils. The Ohmic heating (OH) solenoid was not used but a small inboard coil (part of the shaping coil), providing less than 20% of total poloidal flux, was used. The plasma thus obtained had both internal and edge transport barriers, with an energy confinement time of 1.6 times H-mode scaling, a poloidal beta of 3.6, and a normalized beta of 1.6, and a large bootstrap current fraction (>90%). This result opens up a possibility to reduce, and eventually eliminate, the OH solenoid from a tokamak reactor, which will greatly improve its economic competitiveness. PMID- 14753881 TI - Attractive potential around a thermionically emitting microparticle. AB - We present a simulation study of the charging of a dust grain immersed in a plasma, considering the effect of thermionic electron emission from the grain. It is shown that the orbit motion limited theory is no longer reliable when electron emission becomes large: screening can no longer be treated within the Debye Huckel approach and an attractive potential well can form, leading to the possibility of attractive forces on other grains with the same polarity. We suggest to perform laboratory experiments where emitting dust grains could be used to create nonconventional dust crystals or macromolecules. PMID- 14753882 TI - Kelvin-wave cascade and decay of superfluid turbulence. AB - Kelvin waves (kelvons), the distortion waves on vortex lines, play a key part in the relaxation of superfluid turbulence at low temperatures. We present a weak turbulence theory of kelvons. We show that nontrivial kinetics arises only beyond the local-induction approximation and is governed by three-kelvon collisions; a corresponding kinetic equation is derived. We prove the existence of Kolmogorov cascade and find its spectrum. The qualitative analysis is corroborated by numeric study of the kinetic equation. The application of the results to the theory of superfluid turbulence is discussed. PMID- 14753883 TI - Contact dynamics in a gently vibrated granular pile. AB - We use multispeckle diffusive wave spectroscopy to probe the micron-scale dynamics of a water-saturated granular pile submitted to discrete gentle taps. The typical time scale between plastic events is found to increase dramatically with the number of applied taps. Furthermore, this microscopic dynamics weakly depends on the solid fraction of the sample. This process is largely analogous to the aging phenomenon observed in thermal glassy systems. We propose a heuristic model where this slowing-down mechanism is associated with a slow evolution of the distribution of the contact forces between particles. This model accounts for the main features of the observed dynamics. PMID- 14753884 TI - Cooperative dynamics in two dimensions. AB - We report results from molecular dynamics simulations of cooperative motion in a quasi-two-dimensional system of colloid particles. We find that the onset of the deviation of the single-particle displacement distribution from Gaussian form starts in the liquid phase and extends, with increasing magnitude, through the hexatic phase into the crystalline phase. The time for which the deviation is maximum increases exponentially with the density. As the density increases toward the hexatic phase a third dynamical relaxation mode emerges. We argue that the collective motion is generated by superpositions of instantaneous normal mode vibrations, with lifetimes that increase with the density, along paths with strong bond-orientation correlation. PMID- 14753885 TI - Coherent interactions of terahertz strain solitons and electronic two-level systems in photoexcited ruby. AB - We observe coherent interactions between an ultrashort, longitudinal acoustic soliton train and the 29-cm(-1) electronic transition in photoexcited ruby. Propagation of the strain pulses over millimeter distance through an excited zone reveals striking behavior of the induced electronic population, which has been explained by impulsive excitation of the two-level systems, combined with the nonlinear properties of the solitons in the resonant medium. This opens up new possibilities for coherent manipulation of ultrashort acoustic pulses by local electronic centers. PMID- 14753886 TI - Rejection-free geometric cluster algorithm for complex fluids. AB - We present a novel, generally applicable Monte Carlo algorithm for the simulation of fluid systems. Geometric transformations are used to identify clusters of particles in such a manner that every cluster move is accepted, irrespective of the nature of the pair interactions. The rejection-free and nonlocal nature of the algorithm make it particularly suitable for the efficient simulation of complex fluids with components of widely varying size, such as colloidal mixtures. Compared to conventional simulation algorithms, typical efficiency improvements amount to several orders of magnitude. PMID- 14753887 TI - High-frequency vibrational properties of metallic nanocrystalline grain boundaries. AB - The high-frequency phonon properties of a computer generated nanocrystalline (nc) fcc Ni with a mean grain size of 5 nm are investigated by directly calculating the on-site phonon Green's function using a recursion technique based on a continued fraction representation. It is found that the high-frequency tail, observed in both experiment and previous simulation work, arises primarily from spatially confined vibrational modes forming within the nc grain boundary regions. PMID- 14753888 TI - Saddles in the energy landscape: extensivity and thermodynamic formalism. AB - We formally extend the energy landscape approach for the thermodynamics of liquids to account for saddle points. By considering the extensive nature of macroscopic potential energies, we derive the scaling behavior of saddles with system size, as well as several approximations for the properties of low-order saddles (i.e., those with only a few unstable directions). We then cast the canonical partition function in a saddle-explicit form and develop, for the first time, a rigorous energy landscape approach capable of reproducing trends observed in simulations, in particular, the temperature dependence of the energy and fractional order of sampled saddles. PMID- 14753889 TI - Interaction of silicon dangling bonds with insulating surfaces. AB - We use first principles density functional theory calculations to study the interaction of a model dangling bond silicon tip with the surfaces of CaF2, Al2O3, TiO2, and MgO. In each case the strongest interaction is with the highest anions in the surface. We show that this is due to the onset of chemical bonding with the surface anions, which can be controlled by an electric field across the system. Combining our results and previous studies on semiconductor surfaces suggests that using dangling bond Si tips can provide immediate identification of surface species in atomically resolved noncontact atomic force microscopy and facilitate selective measurements of short-range interactions with surface sites. PMID- 14753890 TI - Gate-voltage dependence of zero-bias anomalies in multiwall carbon nanotubes. AB - Temperature dependence of zero-bias conductance of the vanadium (V)/multiwall carbon nanotube (MWNT)/V structure is studied. As temperature is reduced, the conductance decreases with a functional form consistent with a power law. For the first time, we find that the exponent depends significantly on gate voltage. This exponent dependence cannot be explained by Luttinger-liquid theory for ballistic MWNTs. We interpret the obtained results within the framework of the nonconventional Coulomb blockade theory for strongly disordered MWNTs. PMID- 14753891 TI - Laterally modulated 2D electron system in the extreme quantum limit. AB - We report on magnetotransport of a two-dimensional electron system (2DES), located 32 nm below the surface, with a surface superlattice gate structure of periodicity 39 nm imposing a periodic modulation of its potential. For low Landau level fillings nu, the diagonal resistivity displays a rich pattern of fluctuations, even though the disorder dominates over the periodic modulation. Theoretical arguments based on the combined effects of the long-wavelength, strong disorder and the short-wavelength, weak periodic modulation present in the 2DES qualitatively explain the data. PMID- 14753892 TI - Snell's law for surface electrons: refraction of an electron gas imaged in real space. AB - On NaCl(100)/Cu(111) an interface state band is observed that descends from the surface-state band of the clean copper surface. This band exhibits a Moire pattern-induced one-dimensional band gap, which is accompanied by strong standing wave patterns, as revealed in low-temperature scanning tunneling microscopy images. At NaCl island step edges, one can directly see the refraction of these standing waves, which obey Snell's refraction law. PMID- 14753893 TI - Intrinsic electron accumulation at clean InN surfaces. AB - The electronic structure of clean InN(0001) surfaces has been investigated by high-resolution electron-energy-loss spectroscopy of the conduction band electron plasmon excitations. An intrinsic surface electron accumulation layer is found to exist and is explained in terms of a particularly low Gamma-point conduction band minimum in wurtzite InN. As a result, surface Fermi level pinning high in the conduction band in the vicinity of the Gamma point, but near the average midgap energy, produces charged donor-type surface states with associated downward band bending. Semiclassical dielectric theory simulations of the energy-loss spectra and charge-profile calculations indicate a surface state density of 2.5 (+/ 0.2)x10(13) cm(-2) and a surface Fermi level of 1.64+/-0.10 eV above the valence band maximum. PMID- 14753894 TI - Nonlinear response of a clean one-dimensional wire. AB - We study nonlinear transport in a clean one-dimensional wire fabricated by cleaved edge overgrowth in molecular beam epitaxy. At low electron densities, and with a large applied bias, we observe a feature in the differential conductance similar to the so-called "0.7 structure," found in quantum point contact devices. Using a simple model we suggest a link between this phenomenon, charge neutrality, and unidirectional dynamics in the wire. PMID- 14753895 TI - Signatures of pairing mechanisms and order parameters in ferromagnetic superconductors. AB - Two predictions are made for properties of the ferromagnetic superconductors discovered recently. The first one is that spin-triplet, p-wave pairing in such materials will give the magnons a mass inversely proportional to the square of the magnetization. The second one is based on a specific mechanism for p-wave pairing and predicts that the observed broad anomaly in the specific heat of URhGe will be resolved into a split transition with increasing sample quality. These predictions will help discriminate between different possible mechanisms for ferromagnetic superconductivity. PMID- 14753896 TI - Anisotropic origin of the bending instability of the flux-antiflux interface in type-II superconductors. AB - The physical nature of the macroturbulence in vortex matter in YBCO superconductors is investigated by means of a magneto-optic study of the instability in a single crystal prepared especially for this purpose. The instability develops near those sample edges where the oppositely directed flow of vortices and antivortices, guided by twin boundaries, is characterized by the discontinuity of the tangential component of the hydrodynamic velocity. This fact indicates that the macroturbulence is analogous to the instability of fluid flow at a surface of a tangential velocity discontinuity in classical hydrodynamics and is related to the anisotropic flux motion in the superconductor. PMID- 14753897 TI - Relaxation of hot quasiparticles in a d-wave superconductor. AB - Motivated by recent pump-probe experiments we consider the processes by which "hot" quasiparticles produced near the antinodes of a d-wave superconductor can relax. We show that in a large region of momentum space processes which break Cooper pairs are forbidden by energy and momentum conservation. Equilibration then occurs by scattering with thermal quasiparticles: Umklapp scattering is exponentially suppressed at low temperatures, but small-angle scattering leads to power-law behavior. By solving the Boltzmann equation analytically we make detailed predictions for the temperature and intensity dependence of these processes, which we compare with experiment. PMID- 14753898 TI - Nodal Cooper-pair stabilized phase dynamics in granular d-wave superconductors. AB - Scanning tunneling microscope measurements on single crystals of Bi2Sr2CaCu2O8+x materials have shown that the d-wave superconductivity in cuprates has nanoscale inhomogeneities and is still robust in spite of their presence. We study the dynamics of Josephson coupling between such granular d-wave superconductors, focusing on the effect of nodal Cooper pairs and disorder. We find that the nodal Cooper pairs give rise to a power-law Josephson coupling which leads to the stabilization of the superconducting phase. Our findings suggest that the d-wave superconductivity in an array of grains is unexpectedly robust against a disordering transition, as observed in the experiments. Furthermore, we predict the existence of a planar Josephson-plasmon mode with characteristic frequency that decreases with temperature. PMID- 14753899 TI - Direct transition from Bose glass to normal state in the (K,Ba)BiO3 superconductor. AB - The introduction of columnar defects in (K,Ba )Bi O3 single crystals shifts both the irreversibility and thermodynamic transition lines, respectively, deduced from ac susceptibility (and/or transport) and specific heat measurements, upwards. This shift can be attributed to the defect-induced decrease of the difference (Delta F) between the free energies in the superconducting and the normal states, assuming that the position of the superconducting transition is given by the condition absolute value Delta F approximately k(B )T/xi(3 ). This criterion also perfectly reproduces the influence of the angle between the tracks and the external field. This result suggests that no vortex liquid phase exists in this system. PMID- 14753900 TI - d-Mott phases in one and two dimensions. AB - We use exact diagonalization to determine the spectrum of reduced Hamiltonians based on renormalization group flows to strong coupling. For the half-filled two leg Hubbard ladder we reproduce the known insulating d-Mott ground state with spin and charge gaps. For the saddle point regions of the two-dimensional Hubbard model near half filling we find a crossover to a similar strong coupling state, which truncates the Fermi surface near the saddle points. At lower scales d-wave superconductivity appears on the remaining Fermi surface. PMID- 14753901 TI - Mn interstitial diffusion in (ga,mn)as. AB - We present a combined theoretical and experimental study of the ferromagnetic semiconductor (Ga,Mn)As which explains the remarkably large changes observed on low-temperature annealing. Careful control of the annealing conditions allows us to obtain samples with ferromagnetic transition temperatures up to 159 K. Ab initio calculations, in situ Auger spectroscopy, and resistivity measurements during annealing show that the observed changes are due to out diffusion of Mn interstitials towards the surface, governed by an energy barrier of 0.7-0.8 eV. Electric fields induced by Mn acceptors have a significant effect on the diffusion. PMID- 14753902 TI - Magnetism, charge order, and giant magnetoresistance in SrFeO(3-delta) single crystals. AB - The electronic and magnetic properties of SrFeO(3-delta) single crystals with controlled oxygen content (0< or =delta< or =0.19) have been studied systematically by susceptibility, transport, and spectroscopic techniques. An intimate correlation between the spin-charge ordering and the electronic transport behavior is found. Giant negative as well as positive magnetoresistance are observed. PMID- 14753903 TI - High magnetic field phase diagram of PrOs4Sb12. AB - The magnetic phase diagram of PrOs4Sb12 has been investigated by specific heat measurements between 8 and 32 T. A new Schottky anomaly, due to excitations between two lowest crystalline-electric-field (CEF) singlets, has been found for both H parallel (100) and H parallel (110) above the field where the field induced ordered phase (FIOP) is suppressed. The constructed H-T phase diagram shows weak magnetic anisotropy and implies a crossing of the two CEF levels at about 8-9 T for both field directions. These results provide an unambiguous evidence for the Gamma(1) singlet being the CEF ground state and suggest the level crossing (involving lowest CEF levels) as the driving mechanism of FIOP. PMID- 14753904 TI - First principles calculation of anomalous Hall conductivity in ferromagnetic bcc Fe. AB - We perform a first principles calculation of the anomalous Hall effect in ferromagnetic bcc Fe. Our theory identifies an intrinsic contribution to the anomalous Hall conductivity and relates it to the k-space Berry phase of occupied Bloch states. This dc conductivity has the same origin as the well-known magneto optical effect, and our result accounts for experimental measurement on Fe crystals with no adjustable parameters. PMID- 14753905 TI - Suppression of spin diffusion near a micron-size ferromagnet. AB - We have used the large gradients generated near the ferromagnetic tip of a magnetic resonance force microscope to locally suppress spin diffusion in a silica sample containing paramagnetic electron spins. By controlling the slice location with respect to the tip, the magnetic field gradient was varied from 0.01 to 36 mT/microm, resulting in a fourfold decrease in T-11 and a similar decrease in T(-1)(1 rho). The observed dependence of the relaxation rates on field gradient is consistent with the quenching of flip-flop interactions that mediate the transport of magnetization between slow and fast relaxing spins. PMID- 14753906 TI - Boundary susceptibility in the spin-1/2 chain: Curie-like behavior without magnetic impurities. AB - We investigate the low-temperature thermodynamics of the spin-1/2 Heisenberg chain with open ends. On the basis of boundary conformal field theory arguments and numerical density matrix renormalization group calculations, it is established that in the isotropic case the impurity susceptibility exhibits a Curie-like divergent behavior as the temperature decreases, even in the absence of magnetic impurities. A similar singular temperature dependence is also found in the boundary contributions of the specific heat coefficient. In the anisotropic case, for 1/22(3)P 0,1,2 atomic helium transitions around 1083 nm. AB - We measure the frequency of the 2(3)S1-->2(3)P(0,1,2) transitions of helium in a metastable beam using an optical frequency comb synthesizer. The relative uncertainty of these measurements ranging from 5x10(-11) to 7x10(-12) is, to our knowledge, the most precise result for any optical helium transition. Considering existing accurate values of the 2(3)P fine structure, we measure a centroid value of the 2(3)S-2(3)P frequency of 276 736 495 624.6(2.4) kHz, improving the previous interferometric measurement by 30 times. New accurate values of the 2(3)S-2(3)P and 2(3)P Lamb-shift energies are obtained. PMID- 14753931 TI - Q2 evolution of the neutron spin structure moments using a 3He target. AB - We have measured the spin structure functions g(1) and g(2) of 3He in a double spin experiment by inclusively scattering polarized electrons at energies ranging from 0.862 to 5.058 GeV off a polarized 3He target at a 15.5 degrees scattering angle. Excitation energies covered the resonance and the onset of the deep inelastic regions. We have determined for the first time the Q2 evolution of Gamma(1)(Q2)= integral (1)(0)g(1)(x,Q2)dx, Gamma(2)(Q2)= integral (1)(0)g(2)(x,Q2)dx, and d(2)(Q2)= integral (1)(0)x(2)[2g(1)(x,Q2)+3g(2)(x,Q2)]dx for the neutron in the range 0.1< or =Q2< or =0.9 GeV2 with good precision. Gamma(1)(Q2) displays a smooth variation from high to low Q2. The Burkhardt Cottingham sum rule holds within uncertainties and d(2) is nonzero over the measured range. PMID- 14753934 TI - Collision-induced absorption of hydrogen deuteride dissolved in liquid neon. AB - First observation of the collision-induced IR-fundamental band of hydrogen deuteride (HD) in liquid neon is described. A developed intracell rattling model yields highly accurate fits to the measured diffuse Q1-branch profiles enabling a detailed shape analysis of the adjacent spectral lines. Strong intracollisional anticorrelation found at the location of the permanent dipole-allowed R1(0) transition is interpreted in the frame of the Fano-Mori theory. A new striking narrowing effect (by 30%) is observed on the sharp Q(q)(1)(0) vibrational line with increasing HD concentration in solutions. The T1(0), U1(0) and a few pair transitions are identified in the spectrum. PMID- 14753935 TI - Strong field quantum path control using attosecond pulse trains. AB - We show that attosecond pulse trains have a natural application in the control of strong field processes. In combination with an intense infrared laser field, the pulse train can be used to microscopically select a single quantum path contribution to a process that would otherwise consist of several interfering components. We present calculations that demonstrate this by manipulating the time-frequency properties of high order harmonics at the single atom level. This quantum path selection can also be used to define a high resolution attosecond clock. PMID- 14753936 TI - Filtered optical feedback induced frequency dynamics in semiconductor lasers. AB - We demonstrate experimentally and numerically that, by spectrally filtering the delayed optical feedback into a semiconductor laser, one can elicit novel dynamics in the frequency of the laser output light on a time scale that is set by the delay time of the feedback. In particular, we show that through a judicious choice of the filter bandwidth, and its frequency relative to that of the laser, one can produce controlled oscillations in the frequency of light from the laser. PMID- 14753937 TI - Time-reversal analysis for scatterer characterization. AB - A new application of time-reversal processing of wave scattering data permits characterization of scatterers by analyzing the number and nature of the singular functions (or eigenfunctions) associated with individual scatterers when they have multiple contributions from monopole, dipole, and/or quadrupole scattering terms. We discuss acoustic, elastic, and electromagnetic scattering problems for low frequencies. Specific examples for electromagnetic scattering from one of a number of small conducting spheres show that each sphere can have up to six distinct time-reversal eigenfunctions associated with it. PMID- 14753938 TI - Large deformations during the coalescence of fluid interfaces. AB - Surface forces and shape changes were simultaneously measured during the approach and coalescence of two liquid-liquid and liquid-air interfaces. Large normal and lateral deformations were observed that are nevertheless consistent with a simple theoretical analysis of the long-range effects of short-range attractive van der Waals forces. The results imply that two fluidlike structures such as liquid droplets and soft biological cells can sense each other at much larger separations than previously assumed based on criteria taken from the interactions of hard particles. PMID- 14753939 TI - Electron thermal diffusivity due to the electron temperature gradient mode. AB - Charge neutrality breaks down in the short wavelength toroidal electron temperature gradient mode. In contrast to the ion temperature gradient mode, the wave number normalized by the Debye wave number, k/k(De), appears as a natural scale parameter, rather than the finite Larmor radius parameter k( perpendicular )rho(e). The growth rate and consequent mixing length estimate yields an electron thermal diffusivity large enough to be relevant to tokamaks. PMID- 14753940 TI - Effect of collisional zonal-flow damping on flux-driven turbulent transport. AB - The effect of collisional damping of zonal flows (ZFs) on ion-temperature gradient (ITG) driven turbulence in a toroidal plasma is investigated by means of a 3D global fluid model with flux boundary conditions. Results from simulations show an increase of the energy confinement time and a stabilization of turbulence with the inverse of the collisionality nu(*). The stabilization mechanism is identified as an effect of the increased shearing rate of ZFs, which shift upwards the ITG turbulence effective threshold. The shearing rate of ZFs is also seen to depend on the injected power. As a consequence, the effective heat conductivity depends parametrically on the input power. PMID- 14753941 TI - Gapless spin liquid behavior in two-dimensional solid 3He. AB - Direct demagnetization has been made for two-dimensional solid 3He in both the paramagnetic and the antiferromagnetic phases. The lowest temperature is about 10 microK, judging from the observed magnetization for the paramagnetic solid 3He. The magnetization of the antiferromagnetic solid 3He shows a gradual increase to about 10 microK for the 4/7 phase adsorbed on both one layer of 4He and two layers of HD preplated graphite. This strongly suggests that the triangular antiferromagnet with the higher order multiple exchange has a quantum spin liquid ground state with nearly zero or extremely small spin gap less than 10 microK. PMID- 14753942 TI - Entropy-stabilized smectic C phase in a system of zigzag-shaped molecules. AB - We report Monte Carlo simulations of a system of rigid zigzag-shaped molecules that demonstrate that simple excluded-volume interactions are sufficient to produce a fluid tilted lamellar [smectic C (SmC)] liquid crystal phase. The molecules are composed of three rigidly linked hard spherocylinders arranged in a zigzag fashion. By varying the zigzag angle we have mapped out the whole phase diagram as a function of pressure and zigzag angle Psi. For Psi between 35 degrees and 80 degrees our model simulation exhibits the SmC phase. This is the first conclusive evidence where steric interactions arising out of molecular shape alone induce the occurrence of the SmC phase for a wide range of zigzag angles. For smaller Psi, a transition from tilted crystal to crystal is observed. PMID- 14753943 TI - Local strain-mediated chemical potential control of quantum dot self-organization in heteroepitaxy. AB - From observations of self-assembly of Ge quantum dots directed by substrate morphology, we propose the concept of control of ordering in heteroepitaxy by a local strain-mediated surface chemical potential. Using quite simple lithography, we demonstrate directed quantum dot ordering. The strain part of the chemical potential is caused by the spatially nonuniform relaxation of the strained layer, which in our study is the Ge wetting layer, but, more generally, can be a deposited strained buffer layer. This model provides a consistent picture of prior literature. PMID- 14753944 TI - High frequency dynamics in a monatomic glass. AB - The high frequency dynamics of glassy selenium has been studied by inelastic x ray scattering at beam line BL35XU (SPring-8). The high quality of the data allows one to pinpoint the existence of a dispersing acoustic mode for wave vectors (Q) of 1.5>1). We find that the system's conductance and differential capacitance exhibits activated behavior, approximately exp([-T(*)/T]. The gap T(*) represents the energy needed to create a long single-electron charge soliton propagating through the array. This scale is parametrically larger than the energy at which conventional perturbation theory breaks down. PMID- 14753952 TI - Scanning tunneling microscopy chemical signature of point defects on the MoS2(0001) surface. AB - Using ab initio calculations, we have studied the modification of the electronic structure of the MoS2(0001) surface by several point defects: a surface S vacancy and different transition metal atoms substituting a S atom (Pd, Au, Fe, and V). With a S vacancy, a gap state appears with weight mostly on the Mo and S atoms surrounding the vacancy. The substitutional atoms of complete d band (Pd and Au) do not present magnetic polarization and slightly modify the DOS near the Fermi energy. On the other hand, the incomplete d band atoms (Fe and V) present spin polarization and modify significantly the states near the band edges. From calculated STM images and STS curves, we show that this chemical signature can be measured and used to characterize the surface defects of the substrate which are suitable nucleation centers for nanocluster growth. PMID- 14753953 TI - Hydrodynamic effects in interacting Fermi electron jets. AB - We theoretically study hydrodynamic phenomena originating from electron-electron collisions in a two-dimensional Fermi system. We demonstrate that an electron beam sweeping past an aperture creates a pumping effect, attracting carriers from this aperture. This pumping effect originates from the specific electric potential distribution induced by the injected electrons. In the regions near the main stream of injected electrons, a positive potential is induced by the injected electrons. Thus, the normally repulsive Coulomb interaction leads to an attractive force in the Fermi system. This quantum pumping mechanism in a Fermi system differs qualitatively from the Bernoulli pumping effect in classical liquids. We also discuss possible experimental realizations. PMID- 14753954 TI - Geometrical dependence of high-bias current in multiwalled carbon nanotubes. AB - We have studied the high-bias transport properties of the different shells that constitute a multiwalled carbon nanotube. The current is shown to be reduced as the shell diameter is decreased or the length is increased. We assign this geometrical dependence to the competition between the electron-phonon scattering process and Zener tunneling. PMID- 14753955 TI - Two-particle Aharonov-Bohm effect and entanglement in the electronic Hanbury Brown-Twiss setup. AB - We analyze a Hanbury Brown-Twiss geometry in which particles are injected from two independent sources into a mesoscopic conductor in the quantum Hall regime. All partial waves end in different reservoirs without generating any single particle interference; in particular, there is no single-particle Aharonov-Bohm effect. However, exchange effects lead to two-particle Aharonov-Bohm oscillations in the zero-frequency current cross correlations. We demonstrate that this is related to two-particle orbital entanglement, detected via violation of a Bell inequality. The transport is along edge states and only adiabatic quantum point contacts and normal reservoirs are employed. PMID- 14753956 TI - Metastable resistance-anisotropy orientation of two-dimensional electrons in high Landau levels. AB - In half-filled high Landau levels, two-dimensional electron systems possess collective phases which exhibit a strongly anisotropic resistivity tensor. A weak, but as yet unknown, rotational symmetry-breaking potential native to the host semiconductor structure is necessary to orient these phases in macroscopic samples. Making use of the known external symmetry-breaking effect of an in-plane magnetic field, we find that the native potential can have two orthogonal local minima. It is possible to initialize the system in the higher minimum and then observe its relaxation toward equilibrium. PMID- 14753957 TI - Bosonization for disordered and chaotic systems. AB - Using a supersymmetry formalism, we reduce exactly the problem of electron motion in an external potential to a new supermatrix model valid at all distances. All approximate nonlinear sigma models obtained previously for disordered systems can be derived from our exact model using a coarse-graining procedure. As an example, we consider a model for a smooth disorder and demonstrate that using our approach does not lead to a "mode-locking" problem. As a new application, we consider scattering on strong impurities for which the Born approximation cannot be used. Our method provides a new calculational scheme for disordered and chaotic systems. PMID- 14753958 TI - Effect of quantum confinement on the dielectric function of PbSe. AB - Monolayers of lead selenide nanocrystals of a few nanometers in height have been made by electrodeposition on a Au(111) substrate. These layers show a thickness dependent dielectric function, which was determined using spectroscopic ellipsometry. The experimental results are compared with electronic structure calculations of the imaginary part of the dielectric function of PbSe nanocrystals. We demonstrate that the size-dependent variation of the dielectric function is affected by quantum confinement at well-identifiable points in the Brillouin zone, different from the position of the band-gap transition. PMID- 14753959 TI - Transport in ultraclean YBa2Cu3O7: neither unitary nor born impurity scattering. AB - The thermal conductivity of ultraclean YBa2Cu3O7 was measured at very low temperature in magnetic fields up to 13 T. The temperature and field dependence of the electronic heat conductivity show that two widespread assumptions of transport theory applied to unconventional superconductors fail for clean cuprates: impurity scattering cannot be treated in the usual unitary limit (nor indeed in the Born limit), and scattering of quasiparticles off vortices cannot be neglected. Our study also sheds light on the long-standing puzzle of a sudden onset of a "plateau" in the thermal conductivity of Bi-2212 versus field. PMID- 14753960 TI - Scaling of the magnetic response in doped antiferromagnets. AB - A theory of the anomalous omega/T scaling of the dynamic magnetic response in cuprates at low doping is presented. It is based on the memory function representation of the dynamical spin susceptibility in a doped antiferromagnet where the damping of the collective mode is constant and large, whereas the equal time spin correlations saturate at low T. Exact diagonalization results within the t-J model are shown to support assumptions. Consequences, for both the scaling function and the normalization amplitude, are well in agreement with neutron scattering results. PMID- 14753961 TI - Heavy fermion superconductivity and magnetic order in noncentrosymmetric CePt3Si. AB - CePt3Si is a novel heavy fermion superconductor, crystallizing in the CePt3B structure as a tetragonally distorted low symmetry variant of the AuCu3 structure type. CePt3Si exhibits antiferromagnetic order at T(N) approximately 2.2 K and enters into a heavy fermion superconducting state at T(c) approximately 0.75 K. Large values of H(')(c2) approximately -8.5 T/K and H(c2)(0) approximately 5 T refer to heavy quasiparticles forming Cooper pairs. Hitherto, CePt3Si is the first heavy fermion superconductor without a center of symmetry. PMID- 14753962 TI - Vortex state in NaxCoO2.yH2O: px+/-ipy-wave versus d(x2-y2)+/-id xy-wave pairing. AB - Based on an effective Hamiltonian specified in the triangular lattice with possible p(x)+/-ip(y)- or dx(2)(-y(2))+/-id(xy)-wave pairing, which has close relevance to the newly discovered Na0.35CoO2.yH(2)O, the electronic structure of the vortex state is studied by solving the Bogoliubov-de Gennes equations. It is found that p(x)+/-ip(y) wave is favored for the electron doping as the hopping integral t<0. The lowest-lying vortex bound states are found to have, respectively, zero and positive energies for p(x)+/-ip(y)- and dx(2)(-y(2))+/ id(xy)-wave superconductors, whose vortex structures exhibit the intriguing sixfold symmetry. In the presence of strong on-site repulsion, the antiferromagnetic order and local ferromagnetic moment are induced around the vortex cores for the former and the latter, respectively, both of which cause the splitting of the local density of states peaks due to the lifting of spin degeneracy. PMID- 14753963 TI - Kinetic energy driven pairing in cuprate superconductors. AB - Pairing occurs in conventional superconductors through a reduction of the electronic potential energy accompanied by an increase in kinetic energy. In the underdoped cuprates, optical experiments show that pairing is driven by a reduction of the electronic kinetic energy. Using the dynamical cluster approximation we study superconductivity in the two-dimensional Hubbard model. We find that pairing is indeed driven by the kinetic energy and that superconductivity evolves from an unconventional state with partial spin-charge separation, to a superconducting state with quasiparticle excitations. PMID- 14753964 TI - Direct-current induced dynamics in Co90 Fe10/Ni80 Fe20 point contacts. AB - We have directly measured coherent high-frequency magnetization dynamics in ferromagnetic films induced by a spin-polarized dc current. The precession frequency can be tuned over a range of several gigahertz by varying the applied current. The frequencies of excitation also vary with applied field, resulting in a microwave oscillator that can be tuned from below 5 to above 40 GHz. This novel method of inducing high-frequency dynamics yields oscillations having quality factors from 200 to 800. We compare our results with those from single-domain simulations of current-induced dynamics. PMID- 14753965 TI - Optical-spectroscopic detection of spin-exchange interaction pyrochlore molybdates. AB - Optical spectroscopy was used to determine nearest-neighbor spin correlations in pyrochlore molybdates R2Mo2O7 (R=Y, Sm, and Nd), which exhibit ferromagnetic metal to spin-glass insulator transition as the R ion size decreases. Using an analysis based on the orbitally degenerate Hubbard model, we could estimate important physical parameters, such as the effective on-site Coulomb energy U(eff) and the Hund rule exchange coupling J(H). We demonstrated experimentally and theoretically that the effective superexchange interaction between the Mo ions depends on J(H)/U(eff), which determines the phase boundary of the magnetic ground states. PMID- 14753966 TI - Anomalous fluctuations in phases with a broken continuous symmetry. AB - It is shown that the Goldstone modes associated with a broken continuous symmetry lead to anomalously large fluctuations of the zero field order parameter at any temperature below T(c). In dimensions 2>1 spins or qubits and study the average entanglement that can be localized between two separated spins by performing local measurements on the other individual spins. We show that all classical correlation functions provide lower bounds to this localizable entanglement, which follows from the observation that classical correlations can always be increased by doing appropriate local measurements on the other qubits. We analyze the localizable entanglement in familiar spin systems and illustrate the results on the hand of the Ising spin model, in which we observe characteristic features for a quantum phase transition such as a diverging entanglement length. PMID- 14753969 TI - Classical capacity of the lossy bosonic channel: the exact solution. AB - The classical capacity of the lossy bosonic channel is calculated exactly. It is shown that its Holevo information is not superadditive, and that a coherent-state encoding achieves capacity. The capacity of far-field, free-space optical communications is given as an example. PMID- 14753970 TI - Transitions to synchrony in coupled bursting neurons. AB - Certain cells in the brain, for example, thalamic neurons during sleep, show spike-burst activity. We study such spike-burst neural activity and the transitions to a synchronized state using a model of coupled bursting neurons. In an electrically coupled network, we show that the increase of coupling strength increases incoherence first and then induces two different transitions to synchronized states, one associated with bursts and the other with spikes. These sequential transitions to synchronized states are determined by the zero crossings of the maximum transverse Lyapunov exponents. These results suggest that synchronization of spike-burst activity is a multi-time-scale phenomenon and burst synchrony is a precursor to spike synchrony. PMID- 14753971 TI - Role of synaptic filtering on the firing response of simple model neurons. AB - During active states of the brain neurons process their afferent currents with an effective membrane time constant much shorter than its value at rest. This fact, together with the existence of several synaptic time scales, determines to which aspects of the input the neuron responds best. Here we present a solution to the response of a leaky integrate-and-fire neuron with synaptic filters when long synaptic times are present, and predict the firing rate for all values of the synaptic time constant. We also discuss under which conditions this neuron becomes a coincidence detector. PMID- 14753972 TI - Fluctuations in network dynamics. AB - Most complex networks serve as conduits for various dynamical processes, ranging from mass transfer by chemical reactions in the cell to packet transfer on the Internet. We collected data on the time dependent activity of five natural and technological networks, finding that for each the coupling of the flux fluctuations with the total flux on individual nodes obeys a unique scaling law. We show that the observed scaling can explain the competition between the system's internal collective dynamics and changes in the external environment, allowing us to predict the relevant scaling exponents. PMID- 14753973 TI - Comment on "dynamic behavior of anisotropic nonequilibrium driving lattice gases". PMID- 14753975 TI - Bohr-Sommerfeld quantization of spin Hamiltonians. AB - The Bohr-Sommerfeld rule for a spin system is obtained, including the first quantum corrections. The rule applies to both integer and half-integer spin. It is tested for various models, in particular, the Lipkin-Meshkov-Glick model, and found to agree very well with exact results. PMID- 14753976 TI - Quantum reversibility: is there an echo? AB - We study the possibility to undo the quantum mechanical evolution in a time reversal experiment. The naive expectation, as reflected in the common terminology ("Loschmidt echo"), is that maximum compensation results if the reversed dynamics extends to the same time as the forward evolution. We challenge this belief and demonstrate that the time t(r) for maximum return probability is in general shorter. We find that t(r) depends on lambda=epsilon(evol)/epsilon(prep), being the ratio of the error in setting the parameters (fields) for the time-reversed evolution to the perturbation which is involved in the preparation process. Our results should be observable in spin echo experiments where the dynamical irreversibility of quantum phases is measured. PMID- 14753977 TI - Pseudo-dirac neutrinos: a challenge for neutrino telescopes. AB - Neutrinos may be pseudo-Dirac states, such that each generation is actually composed of two maximally mixed Majorana neutrinos separated by a tiny mass difference. The usual active neutrino oscillation phenomenology would be unaltered if the pseudo-Dirac splittings are deltam(2) less, similar 10(-12) eV(2); in addition, neutrinoless double beta decay would be highly suppressed. However, it may be possible to distinguish pseudo-Dirac from Dirac neutrinos using high-energy astrophysical neutrinos. By measuring flavor ratios as a function of L/E, mass-squared differences down to deltam(2) approximately 10(-18) eV(2) can be reached. We comment on the possibility of probing cosmological parameters with neutrinos. PMID- 14753978 TI - The first law for slowly evolving horizons. AB - We study the mechanics of Hayward's trapping horizons, taking isolated horizons as equilibrium states. Zeroth and second laws of dynamic horizon mechanics come from the isolated and trapping horizon formalisms, respectively. We derive a dynamical first law by introducing a new perturbative formulation for dynamic horizons in which "slowly evolving" trapping horizons may be viewed as perturbatively nonisolated. PMID- 14753979 TI - Pulsar recoil by large-scale anisotropies in supernova explosions. AB - Assuming that the neutrino luminosity from the neutron star core is sufficiently high to drive supernova explosions by the neutrino-heating mechanism, we show that low-mode (l=1,2) convection can develop from random seed perturbations behind the shock. A slow onset of the explosion is crucial, requiring the core luminosity to vary slowly with time, in contrast to the burstlike exponential decay assumed in previous work. Gravitational and hydrodynamic forces by the globally asymmetric supernova ejecta were found to accelerate the remnant neutron star on a time scale of more than a second to velocities above 500 km s(-1), in agreement with observed pulsar proper motions. PMID- 14753980 TI - Baryogenesis in a flat direction with neither baryon nor lepton charge. AB - We present a new mechanism of spontaneous baryogenesis. Usually such mechanisms require a derivative coupling between a scalar field and baryon current. In our model, the scalar field instead derivatively couples to a current associated with some global symmetry U(1)(Q). In this case, baryogenesis is still possible provided that an interaction exists, which violates the baryon and U(1)(Q) symmetries simultaneously. As a concrete example, we discuss baryogenesis in a flat direction with neither baryon nor lepton charge. PMID- 14753981 TI - J/psi and eta(c) in the deconfined plasma from lattice QCD. AB - Analyzing correlation functions of charmonia at finite temperature (T) on 32(3)x(32-96) anisotropic lattices by the maximum entropy method (MEM), we find that J/psi and eta(c) survive as distinct resonances in the plasma even up to T approximately 1.6T(c) and that they eventually dissociate between 1.6T(c) and 1.9T(c) (T(c) is the critical temperature of deconfinement). This suggests that the deconfined plasma is nonperturbative enough to hold heavy-quark bound states. The importance of having a sufficient number of temporal data points in MEM analyses is also emphasized. PMID- 14753983 TI - Probing quark-distribution amplitudes through generalized parton distributions at large momentum transfer. AB - In the large momentum transfer limit, generalized parton distributions can be calculated through a QCD factorization theorem which involves perturbatively calculable hard kernels and light-cone parton distribution amplitudes of hadrons. We illustrate this through the H(q)(x,xi,t) distribution for the pion and the proton, presenting the hard kernels at leading order. As a result, experimental data on the generalized parton distributions in this regime can be used to determine the functional form of the parton distribution amplitudes which has thus far been quite challenging to obtain. Our result can also be used as a constraint in phenomenological generalized parton distribution parametrizations. PMID- 14753984 TI - Precision measurement of the neutron spin asymmetryA(n)(1) and spin-flavor decomposition in the valence quark region. AB - We have measured the neutron spin asymmetry A(n)(1) with high precision at three kinematics in the deep inelastic region at x=0.33, 0.47, and 0.60, and Q(2)=2.7, 3.5, and 4.8 (GeV/c)(2), respectively. Our results unambiguously show, for the first time, that A(n)(1) crosses zero around x=0.47 and becomes significantly positive at x=0.60. Combined with the world proton data, polarized quark distributions were extracted. Our results, in general, agree with relativistic constituent quark models and with perturbative quantum chromodynamics (PQCD) analyses based on the earlier data. However they deviate from PQCD predictions based on hadron helicity conservation. PMID- 14753986 TI - Probing the molecular environment using spin-resolved photoelectron spectroscopy. AB - Angle- and spin-resolved photoelectron spectroscopy with linearly and circularly polarized synchrotron radiation were used to study the electronic structure of model triatomic molecules, hydrogen sulfide, and carbonyl sulfide. The spin polarization measurements of the molecular field split components of the S 2p photolines revealed a strong effect of the different molecular environments. The validity of simple atomic models to explain the results is discussed. PMID- 14753987 TI - How to use lasers for imaging attosecond dynamics of nuclear processes. AB - We identify a laser configuration in which attosecond electron wave packets are ionized, accelerated to multi-MeV energies, and refocused onto their parent ion. Magnetic focusing of the electron wave packet results in return currents comparable with large scale accelerator facilities. This technique opens an avenue towards imaging attosecond dynamics of nuclear processes. PMID- 14753988 TI - Corrugating and anticorrugating static interactions in helium-atom scattering from metal surfaces. AB - We perform a density-functional-theory calculation of the static repulsive potential of He scattering off a noble and a simple metal surface. The classical turning point of He on Cu(111) is found to be closer to the metal when the adatom is at top than at bridge site (anticorrugating effect). The potential of He on Al(111) is instead corrugated. By comparing the results of the two systems, we conclude that the He-metal anticorrugating effect occurs when the kinetic energy difference for He at top and bridge sites is larger than the electrostatic one, and an induced localized dipole on He is formed. PMID- 14753982 TI - Measurements of the D(sJ) resonance properties. AB - We report measurements of the properties of the D(+)(sJ)(2317) and D(+)(sJ)(2457) resonances produced in continuum e(+)e(-) annihilation near sqrt[s]=10.6 GeV. The analysis is based on an 86.9 fb(-1) data sample collected with the Belle detector at KEKB. We determine the masses to be M(D(+)(sJ)(2317))=2317.2+/-0.5(stat)+/ 0.9(syst) MeV/c(2) and M(D(+)(sJ)(2457))=2456.5+/-1.3(stat)+/-1.3(syst) MeV/c(2). We observe the radiative decay mode D(+)(sJ)(2457)-->D(+)(s)gamma and the dipion decay mode D(+)(sJ)(2457)-->D(+)(s)pi(+)pi(-) and determine their branching fractions. No corresponding decays are observed for the D(sJ)(2317) state. These results are consistent with the spin-parity assignments of 0(+) for the D(sJ)(2317) and 1(+) for the D(sJ)(2457). PMID- 14753985 TI - Flavor decomposition of the sea-quark helicity distributions in the nucleon from semiinclusive deep inelastic scattering. AB - Double-spin asymmetries of semiinclusive cross sections for the production of identified pions and kaons have been measured in deep inelastic scattering of polarized positrons on a polarized deuterium target. Five helicity distributions including those for three sea quark flavors were extracted from these data together with reanalyzed previous data for identified pions from a hydrogen target. These distributions are consistent with zero for all three sea flavors. A recently predicted flavor asymmetry in the polarization of the light quark sea appears to be disfavored by the data. PMID- 14753989 TI - Formation and stability of high-spin alkali clusters. AB - Helium nanodroplet isolation has been applied to agglomerate alkali clusters at temperatures of 380 mK. The very weak binding to the surface of the droplets allows a selection of only weakly bound, high-spin states. Here we show that larger clusters of alkali atoms in high-spin states can be formed. The lack of strong bonds from pairing electrons makes these systems nonmetallic, van der Waals-like complexes of metal atoms. We find that sodium and potassium readily form such clusters containing up to 25 atoms. In contrast, this process is suppressed for rubidium and cesium. Apparently, for these heavy alkalis, larger high-spin aggregates are not stable and depolarize spontaneously upon cluster formation. PMID- 14753990 TI - Interferometric methods to measure orbital and spin, or the total angular momentum of a single photon. AB - We propose interferometric methods capable of measuring either the total angular momentum, or simultaneously measuring the spin and orbital angular momentum of single photons. This development enables the measurement of any angular momentum eigenstate of a single photon. The work allows the investigation of single-photon two-qubit entangled states and has implications for high density information transfer. PMID- 14753991 TI - Discrete entanglement distribution with squeezed light. AB - We show how one can entangle distant atoms by using squeezed light. Entanglement is obtained in steady state, and can be increased by manipulating the atoms locally. We study the effects of imperfections, and show how to scale up the scheme to build a quantum network. PMID- 14753992 TI - Pointwise dimensions of the lorenz attractor. AB - We discuss a connection between two complementary views of the Lorenz attractor: the first is the accepted view where the attractor has a smooth measure on a fractal support. This complex system is alternatively manifest as a self-similar curve for the pointwise dimension alpha. We describe why the latter approach is accessible for the analysis of an experimental signal. PMID- 14753993 TI - Self-affine fronts in self-affine fractures: large and small-scale structure. AB - The evolution and spatial structure of displacement fronts in fractures with self affine rough walls are studied by numerical simulations. The fractures are open and the two faces are identical but shifted along their mean plane, either parallel or perpendicular to the flow. An initially flat front advected by the flow is progressively distorted into a self-affine front with a Hurst exponent equal to that of the fracture walls. The lower cutoff of the self-affine regime depends only on the aperture, while the upper cutoff grows with the lateral shift and linearly with the width of the front. PMID- 14753994 TI - Observation of odd toroidal Alfven eigenmodes. AB - Experimental evidence is presented for the existence of the theoretically predicted odd toroidicity induced Alfven eigenmode (TAE) from the simultaneous appearance of odd and even TAEs in a normal shear discharge of the joint European torus. The modes are observed in low central magnetic shear plasmas created by injecting lower hybrid current drive. A fast ion population was created by applying ion cyclotron heating at the high-field side to excite the TAEs. The odd TAEs were identified from their frequency, mode number, and timing relative to the even TAEs. PMID- 14753995 TI - Proton shock acceleration in laser-plasma interactions. AB - The formation of strong, high Mach number (2-3), electrostatic shocks by laser pulses incident on overdense plasma slabs is observed in one- and two-dimensional particle-in-cell simulations, for a wide range of intensities, pulse durations, target thicknesses, and densities. The shocks propagate undisturbed across the plasma, accelerating the ions (protons). For a dimensionless field strength parameter a(0)=16 (Ilambda(2) approximately 3 x 10(20) W cm(-2) microm(2), where I is the intensity and lambda the wavelength), and target thicknesses of a few microns, the shock is responsible for the highest energy protons. A plateau in the ion spectrum provides a direct signature for shock acceleration. PMID- 14753996 TI - Twist elasticity and anchoring in a lamellar nematic phase. AB - Electro-optic measurements were performed on a lamellar nematic phase in which the mesogenic moieties lie in lamellae that are separated by partially perfluorinated side groups. The twist elastic constant K22, viscosity gamma(1), and the quadratic and quartic anchoring strength coefficients are reported. K22 and gamma(1) are found to be considerably smaller than that of typical three dimensional nematics. The small K22 is due to the greatly weakened interactions between the spatially separated lamellae. PMID- 14753997 TI - Identification of the carbon dangling bond center at the 4H-SiC/SiO(2) interface by an EPR study in oxidized porous SiC. AB - We report the observation of a paramagnetic interface defect in thermally oxidized porous n-type doped 4H-SiC/SiO(2). Based on its axial symmetry and resolved hyperfine interactions it is attributed to an sp(3) carbon dangling bond center situated at the SiC side of the interface. This center is electrically active and pins the Fermi level in the oxidized samples. No silicon related paramagnetic dangling bond centers are observed. The formation of dangling bond centers seems to be related to interstitial oxygen diffusion at the interface during the oxidation process. PMID- 14753998 TI - Spatiotemporal behavior of void collapse in shocked solids. AB - Molecular dynamics simulations on a three-dimensional defective Lennard-Jones solid containing a void are performed in order to investigate detailed properties of hot spot generation. In addition to the temperature, I monitor the number of energetically colliding particles which characterizes the intensity of shock enhanced chemistry. This quantity normalized by void volume is found to saturate for nanoscale voids and to be maximized after voids have completely collapsed. It makes an apparent comparison to the temperature which requires much larger void for the enhancement and becomes maximum during the early stage of the collapse. It is also found that the average velocity and the temperature of ejected molecules inside a cubic void are enhanced during the collapse because of the focusing of momentum and energy towards the centerline of a void. PMID- 14753999 TI - Role of mobile interstitial oxygen atoms in defect processes in oxides: interconversion between oxygen-associated defects in SiO(2) glass. AB - The role of mobile interstitial oxygen atoms (O(0)) in defect processes in oxides is demonstrated by interconversion between the oxygen dangling bond and the peroxy radical (POR) in SiO2 glass. Superstoichiometric O(0) was created by F2 laser photolysis of the interstitial O2. On annealing above 300 degrees C, O(0) migrated and converted the oxygen dangling bond to POR. Exposure to 5.0 eV light converted POR back to a pair of the oxygen dangling bond and O(0) (quantum yield: approximately 0.1). These findings suggest that various defect processes typically occurring in SiO2 glass at approximately 300-500 degrees C are related to migration of O(0), which exists in the glass network in the peroxy linkage form. PMID- 14754000 TI - Low-temperature nucleation in a kinetic Ising model with soft stochastic dynamics. AB - We study low-temperature nucleation in kinetic Ising models by analytical and simulational methods, confirming the general result for the average metastable lifetime, =Aexp((betagamma) (beta=1/k(B)T) [Commun. Math. Phys. 137, 209 (1991)]]. Contrary to common belief, we find that both A and Gamma depend significantly on the stochastic dynamic. In particular, for a "soft" dynamic, in which the effects of the interactions and the applied field factorize in the transition rates, Gamma does not simply equal the energy barrier against nucleation, as it does for the standard Glauber dynamic, which does not have this factorization property. PMID- 14754001 TI - Rigorous analysis of singularities and absence of analytic continuation at first order phase-transition points in lattice-spin models. AB - We report about two new rigorous results on the nonanalytic properties of thermodynamic potentials at first-order phase transition. For lattice models (d>or=2) with arbitrary finite state space, finite-range interactions which have two ground states, we prove that the pressure has no analytic continuation at the first-order phase-transition point, under the only further assumptions that the Peierls condition is satisfied for the ground states and that the temperature is sufficiently low. For Ising models with Kac potentials J(gamma)(x)=gamma(d)phi(gammax), where 00) and analyticity in the mean field limit (gamma SE pointing arrow 0). PMID- 14754002 TI - Superfluid-insulator transition in commensurate disordered bosonic systems: large scale worm algorithm simulations. AB - We report results of large-scale Monte Carlo simulations of superfluid-insulator transitions in disordered commensurate 2D bosonic systems. In the off-diagonal disorder case, we find that the transition is to a gapless incompressible insulator, and its dynamical critical exponent is z=1.5(2). In the diagonal disorder case, we prove the conjecture that rare statistical fluctuations are inseparable from critical fluctuations on the largest scales and ultimately result in crossover to the generic universality class (apparently with z=2). However, even at strong disorder, the universal behavior sets in only at very large space-time distances. This explains why previous studies of smaller clusters mimicked a direct superfluid-Mott-insulator transition. PMID- 14754003 TI - Thermal contraction of carbon fullerenes and nanotubes. AB - We perform molecular dynamics simulations to study shape changes of carbon fullerenes and nanotubes with increasing temperature. At moderate temperatures, these systems gain structural and vibrational entropy by exploring the configurational space at little energy cost. We find that the soft phonon modes, which couple most strongly to the shape, maintain the surface area of these hollow nanostructures. In nanotubes, the gain in entropy translates into a longitudinal contraction, which reaches a maximum at T approximately 800 K. Only at much higher temperatures do the anharmonicities in the vibration modes cause an overall expansion. PMID- 14754004 TI - Creation of an atomic superlattice by immersing metallic adatoms in a two dimensional electron sea. AB - Cerium adatoms, deposited on a Ag(111) surface, are found by low-temperature scanning tunneling microscopy to self-assemble into large ordered hexagonal arrays covering macroscopically the entire surface. We show that the 32 A periodicity of the superlattice is caused by the interaction of surface-state electrons with Ce adatoms and that the large-scale formation of the superlattice is governed by a subtle balance between the sample temperature, the surface diffusion barrier, and the concentration-dependent adatom interaction potential. PMID- 14754005 TI - Two fluid description of the Kondo lattice. AB - We present a two-fluid description of the Kondo lattice that is based on an analysis of CeCoIn5 at various levels of dilution with La. We show that thermal and transport measurements provide evidence for the survival of the Kondo impurity component in the lattice as T-->0 K, and that the evolution of the low temperature properties of the Kondo lattice can be viewed as the partial condensation of the lattice of Kondo centers into a heavy fermion fluid. The resulting two-fluid model is shown to be applicable to the general problem of the ground state of the Kondo lattice. PMID- 14754006 TI - Predicted electric field near small superconducting ellipsoids. AB - We predict the existence of large electric fields near the surface of superconducting bodies of ellipsoidal shape of dimensions comparable to the penetration depth. The electric field is quadrupolar in nature with significant corrections from higher order multipoles. Prolate (oblate) superconducting ellipsoids are predicted to exhibit fields consistent with negative (positive) quadrupole moments, reflecting the fundamental charge asymmetry of matter. PMID- 14754007 TI - Fibrillar templates and soft phases in systems with short-range dipolar and long range interactions. AB - We analyze the thermal fluctuations of particles that have a short-range dipolar attraction and a long-range repulsion. In an inhomogeneous particle density region, or "soft phase," filamentary patterns appear which are destroyed only at very high temperatures. The filaments act as a fluctuating template for correlated percolation in which low-energy excitations can move through the stable pattern by local rearrangements. At intermediate temperatures, dynamically averaged checkerboard states appear. We discuss possible implications for doped cuprate oxides and related materials. PMID- 14754008 TI - Topological oscillations of the magnetoconductance in disordered GaAs layers. AB - Oscillatory variations of the diagonal (Gxx) and Hall (G(xy)) magnetoconductances are discussed in view of topological scaling effects giving rise to the quantum Hall effect. They occur in a field range without oscillations of the density of states due to Landau quantization, and are, therefore, totally different from the Shubnikov-de Haas oscillations. Such oscillations are experimentally observed in disordered GaAs layers in the extreme quantum limit of applied magnetic field with a good description by the unified scaling theory of the integer and fractional quantum Hall effect. PMID- 14754009 TI - Step-lattice-induced band-gap opening at the fermi level. AB - The interaction of the Shockley surface state with the step lattice of vicinal Cu(111) leads to the formation of an electronic superlattice state. On Cu(443), where the average terrace length forms a "shape resonance" with the Fermi wavelength, we find a step-lattice-induced band-gap opening at the Fermi level. A gap magnitude >200 meV is inferred from high resolution photoemission experiments and line shape analysis. The corresponding energy gain with respect to a gapless case is approximately 11 meV/unit cell, and is a substantial contribution to the stabilization of the step lattice. PMID- 14754010 TI - Quasiparticle decoherence in d-wave superconducting qubits. AB - It is usually argued that the presence of gapless quasiparticle excitations at the nodes of the d-wave superconducting gap should strongly decohere the quantum states of a d-wave qubit, making quantum effects practically unobservable. Using a self-consistent linear response nonequilibrium quasiclassical formalism, we show that this is not necessarily true. We find quasiparticle conductance of a d wave grain boundary junction to be strongly phase dependent. Midgap states as well as nodal quasiparticles contribute to the conductance and therefore decoherence. Quantum behavior is estimated to be detectable in a qubit containing a d-wave junction with appropriate parameters. PMID- 14754011 TI - Effects of a collective spin resonance mode on the scanning tunneling microscopy spectra of d-wave superconductors. AB - A high-energy spin resonance mode is known to exist in many high-temperature superconductors. Motivated by recent scanning tunneling microscopy experiments in superconducting Bi(2)Sr(2)CaCu(2)O(8+delta), we study the effects of this resonance mode on the local density of states (LDOS). The coupling between the electrons in a d-wave superconductor and the resonance mode produces high-energy peaks in the LDOS, which displays a two-unit-cell periodic modulation around a nonmagnetic impurity. This suggests a new means to not only detect the dynamical spin collective mode but also study its coupling to electronic excitations. PMID- 14754012 TI - Proximity effect in gold-coated YBa(2)Cu(3)O(7-delta) films studied by scanning tunneling spectroscopy. AB - Scanning tunneling spectroscopy on gold layers overcoating c-axis YBa(2)Cu(3)O(7 delta) (YBCO) films reveals proximity-induced gap structures. The gap size reduces exponentially with the distance from a-axis facets, indicating that the proximity effect is primarily due to the (100) YBCO facets. The penetration depth of superconductivity into the gold is approximately 30 nm, in good agreement with estimations for the dirty limit. The extrapolated gap at the interface is approximately 15 meV, similar to the value of an s-wave component of the order parameter measured at the YBCO surface in recent point-contact experiments. PMID- 14754013 TI - Superconductor-proximity effect in hybrid structures: fractality versus chaos. AB - We study the proximity effect of a superconductor to a normal system with a fractal spectrum. We find that there is no gap in the excitation spectrum, even in the case where the underlying classical dynamics of the normal system is chaotic. An analytical expression for the distribution of the smallest excitation eigenvalue E1 of the hybrid structure is obtained. On small scales it decays algebraically as P(E1) approximately E1(-D0), where D0 is the fractal dimension of the spectrum of the normal system. Our theoretical predictions are verified by numerical calculations performed for various models. PMID- 14754014 TI - Three-dimensional noncollinear antiferromagnetic order in single-crystalline FeMn ultrathin films. AB - We present experimental evidence for a three-dimensional noncollinear antiferromagnetic spin structure in ultrathin single-crystalline fcc Fe50Mn50 layers using magnetic circular dichroism photoelectron emission microscopy and x ray magnetic linear dichroism. Layer-resolved as-grown domain images of epitaxial trilayers grown on Cu(001) in which FeMn is sandwiched between ferromagnetic layers with different easy axes reveal the presence of antiferromagnetic spin components in the film plane and normal to the film plane. An FeMn spin structure with no collinear order in the film plane is consistent with the absence of x-ray magnetic linear dichroism in Fe L3 absorption in FeMn/Co bilayers. PMID- 14754015 TI - Quantum theory of spin dynamics of exciton-polaritons in microcavities. AB - We present the quantum theory of momentum and spin relaxation of exciton polaritons in microcavities. We show that giant longitudinal-transverse splitting of the polaritons mixes their spin states, which results in beats between right- and left-circularly polarized photoluminescence of microcavities, as was recently experimentally observed [Phys. Rev. Lett. 89, 077402 (2002)]]. This effect is strongly sensitive to the bosonic stimulation of polariton scattering. PMID- 14754016 TI - Shallow donors in diamond: chalcogens, pnictogens, and their hydrogen complexes. AB - The utility of diamond as an electronic material is compromised by the lack of a suitable shallow donor. Here, ab initio theory is used to investigate the donor levels of substitutional pnictogen (N, P, As, and Sb) and chalcogen (S, Se, and Te) impurities and chalcogen-hydrogen defects in diamond. Substitutional S is found to be a deep donor, while As and Sb possess donor levels significantly shallower than P, which so far is the most effective shallow donor found by experiment. PMID- 14754017 TI - Ultrafast spectroscopy of excitons in single-walled carbon nanotubes. AB - We studied the femtosecond dynamics of photoexcitations in films containing semiconducting and metallic single-walled carbon nanotubes (SWNTs), using various pump-probe wavelengths and intensities. We found that confined excitons and charge carriers with subpicosecond dynamics dominate the ultrafast response in semiconducting and metallic SWNTs, respectively. Surprisingly, we also found from the exciton excited state absorption bands and multiphoton absorption resonances in the semiconducting nanotubes that transitions between subbands are allowed; this unravels the important role of electron-electron interaction in SWNT optics. PMID- 14754018 TI - Surface miller index dependence of Auger neutralization of ions on surfaces. AB - Neutralization of He+ ions in grazing incidence scattering on Ag(111) and Ag(110) surfaces is studied. These measurements reveal the existence of an order of magnitude difference in the probability of ion survival on Ag(110) and Ag(111). The experimental results are discussed in terms of survival from Auger neutralization, whose rates are derived theoretically. Molecular dynamics simulation of scattered ion trajectories is performed and the surviving ion fractions are then calculated using the theoretical Auger neutralization rates, without adjustable parameters. The calculations agree quite well with the experimental data and show that the observed differences in the neutralization probabilities on these surfaces are related to different extensions of the electron density beyond the surface, resulting from different atomic packing. PMID- 14754020 TI - Robust polarization-based quantum key distribution over a collective-noise channel. AB - We present two polarization-based protocols for quantum key distribution. The protocols encode key bits in noiseless subspaces or subsystems and so can function over a quantum channel subjected to an arbitrary degree of collective noise, as occurs, for instance, due to rotation of polarizations in an optical fiber. These protocols can be implemented using only entangled photon-pair sources, single-photon rotations, and single-photon detectors. Thus, our proposals offer practical and realistic alternatives to existing schemes for quantum key distribution over optical fibers without resorting to interferometry or two-way quantum communication, thereby circumventing, respectively, the need for high precision timing and the threat of Trojan horse attacks. PMID- 14754019 TI - Dome-shaped magnetic phase diagram of thermoelectric layered cobaltites. AB - Using muon spin spectroscopy we have found that, for both NaxCoO(2) (0.6187Os are determined by dating iron meteorites may constrain the possible time dependence of the decay rate and hence of the fine-structure constant alpha, not directly but only in a model-dependent manner. From this point of view, some of the attempts to analyze the Oklo constraint and the results of the quasistellar object absorption lines are reexamined. PMID- 14754040 TI - High statistics measurement of the K+-->pi0e+nu (K+e3) branching ratio. AB - E865 at the Brookhaven National Laboratory AGS collected about 70 000 K(+)(e3) events to measure the K(+)(e3) branching ratio relative to the observed K+- >pi(+)pi(0), K+-->pi(0)micro(+)nu, and K+-->pi(+)pi(0)pi(0) decays. The pi(0) in all the decays was detected using the e(+)e(-) pair from pi(0)-->e(+)e(-)gamma decay and no photons were required. Using the 2002 Particle Data Group branching ratios for the normalization decays, we obtain BR(K(+)(e3(gamma)))=(5.13+/ 0.02(stat)+/-0.09(syst)+/-0.04(norm))%, where K(+)(e3(gamma)) includes the effect of virtual and real photons. This result is approximately 2.3sigma higher than the current Particle Data Group value. Implications for the V(us) element of the CKM matrix, and the matrix's unitarity are discussed. PMID- 14754037 TI - Observation of B-->K*l+l-. AB - We report the observation of the flavor-changing neutral current decay B- >K(*)l(+)l(-) and an im-proved measurement of the decay B-->Kl(+)l(-), where l represents an electron or a muon, with a data sample of 140 fb(-1) accumulated at the Upsilon(4S) resonance with the Belle detector at KEKB. The results for the branching fractions are B(B-->K(*)l(+)l(-))=(11.5(+2.6)(-2.4)+/-0.8+/-0.2)x10(-7) and B(B-->Kl(+)l(-))=(4.8(+1.0)(-0.9)+/-0.3+/-0.1)x10(-7), where the first error is statistical, the second is systematic and the third is from model dependence. PMID- 14754039 TI - Evidence for B0-->pi0pi0. AB - We report evidence for the decay B0-->pi(0)pi(0). The analysis is based on a data sample of 152x10(6) BBmacr; pairs collected at the Upsilon(4S) resonance with the Belle detector at the KEKB e(+)e(-) storage ring. We detect a signal for B0- >pi(0)pi(0) with a significance of 3.4 standard deviations, and measure the branching fraction to be [1.7+/-0.6(stat)+/-0.2(syst)]x10(-6). PMID- 14754038 TI - Measurement of time-dependent CP-violating asymmetries in B0-->phiK(0)S, K+K K0(S), and eta'K0(S) decays. AB - We present an improved measurement of CP-violation parameters in B0-->phiK(0)(S), K(+)K(-)K(0)(S), and eta(')K(0)(S) decays based on a 140 fb(-1) data sample collected at the Upsilon(4S) resonance with the Belle detector at the KEKB energy asymmetric e(+)e(-) collider. One neutral B meson is fully reconstructed in one of the specified decay channels, and the flavor of the accompanying B meson is identified from its decay products. CP-violation parameters for each of the three modes are obtained from the asymmetries in the distributions of the proper-time intervals between the two B decays. We find that the observed CP asymmetry in the B-->phiK(0)(S) decay differs from the standard model (SM) expectation by 3.5 standard deviations, while the other cases are consistent with the SM. PMID- 14754041 TI - Observation of a narrow charmoniumlike state in exclusive B+/--->K+/-pi+pi-J/psi decays. AB - We report the observation of a narrow charmoniumlike state produced in the exclusive decay process B+/--->K+/-pi(+)pi(-)J/psi. This state, which decays into pi(+)pi(-)J/psi, has a mass of 3872.0+/-0.6(stat)+/-0.5(syst) MeV, a value that is very near the M(D0)+M(D(*0)) mass threshold. The results are based on an analysis of 152M B-Bmacr; events collected at the Upsilon(4S) resonance in the Belle detector at the KEKB collider. The signal has a statistical significance that is in excess of 10sigma. PMID- 14754045 TI - Spin symmetry in the antinucleon spectrum. AB - We discuss spin and pseudospin symmetry in the spectrum of single nucleons and single antinucleons in a nucleus. As an example we use relativistic mean field theory to investigate single antinucleon spectra. We find a very well developed spin symmetry in single antineutron and single antiproton spectra. The dominant components of the wave functions of the spin doublet are almost identical. This spin symmetry in antiparticle spectra and the pseudospin symmetry in particle spectra have the same origin. However, it turns out that the spin symmetry in antinucleon spectra is much better developed than the pseudospin symmetry in normal nuclear single particle spectra. PMID- 14754046 TI - Spin-isospin resonances and the neutron skin of nuclei. AB - The Gamow-Teller resonances (GTR) and isobaric analog states (IAS) of a sequence of even-even Sn target nuclei are calculated by using the framework of the relativistic Hartree-Bogoliubov model plus proton-neutron quasiparticle random phase approximation. The calculation reproduces the experimental data on ground state properties, as well as the excitation energies of the isovector excitations. It is shown that the isotopic dependence of the energy spacings between the GTR and IAS provides direct information on the evolution of neutron skin thickness along the Sn isotopic chain. A new method is suggested for determining the difference between the radii of the neutron and proton density distributions along an isotopic chain, based on measurement of the excitation energies of the GTR relative to the IAS. PMID- 14754047 TI - Accurate Rydberg excitations from the local density approximation. AB - Despite the incorrect asymptotic behavior of its potential, the time-dependent local density approximation can yield accurate optical spectra. The oscillator strengths of Rydberg excitations appear in the calculated spectrum as continuum contributions with excellent optical intensity. We explain why, illustrate this for the neon and helium atoms, and also discuss when such calculations of the optical response will be inaccurate. PMID- 14754048 TI - Reevaluation of the use of photoelectron angular distributions as a probe of dynamical processes: strong dependence of such distributions from s1 paradifluorobenzene on photoelectron kinetic energy. AB - Photoelectron angular distributions (PADs) have been measured following the excitation of the S1 origin band in paradifluorobenzene using a range of ionizing wavelengths and for resolved ion vibrational states. The PADs show a dramatic sensitivity to the photoelectron kinetic energy over an energy range of at least 1 eV from threshold, and almost no sensitivity to any prepared intermediate state alignment. This has important consequences for those studies of intramolecular dynamics that use PADs. We suggest that the observed behavior is caused by a shape resonance in the continuum. PMID- 14754042 TI - Observation of the D(sJ)(2317) and D(sJ)(2457) in B decays. AB - We report the first observation of the B-->Dmacr;D(sJ)(2317) and B- >Dmacr;D(sJ)(2457) decays based on 123.8x10(6) BBmacr; events collected with the Belle detector at KEKB. We observe the D(sJ)(2317) decay to D(s)pi(0) and the D(sJ)(2457) decay to the D(*)(s)pi(0) and D(s)gamma final states. We also set 90% C.L. upper limits for the decays D(sJ)(2317)-->D(*)(s)gamma, D(sJ)(2457)- >D(*)(s)gamma, D(sJ)(2457)-->D(s)pi(0), and D(sJ)(2457)-->D(s)pi(+)pi(-). PMID- 14754049 TI - Search for CPT-odd decays of positronium. AB - We have limited a CPT-violating correlation in annihilations of polarized ortho positronium. We searched for an asymmetry in the triple correlation s-->.(k- >(1)xk-->(2)), where k-->(1) and k-->(2) are the two largest photon momenta, and s--> is the spin of the positronium. Using the Gammasphere array of Compton suppressed high-purity germanium detectors, we detected 2.65x10(7) events of ortho-Ps annihilation. The amplitude of a CPT-violating asymmetry in the data set is found to be 0.0026+/-0.0031, a factor of 6 smaller than previous experiments. PMID- 14754050 TI - Semiclassical description of chaos-assisted tunneling. AB - We study tunneling between regular and chaotic regions in the phase space of Hamiltonian systems. We analytically calculate the transition rate and show that its variation depends only on corresponding phase space area and in this sense is universal. We derive the distribution of level splittings associated with the pairs of quasidegenerate regular eigenstates which in the general case is different from a Cauchy distribution. We show that chaos-assisted tunneling leads to level repulsion between regular eigenstates, solving the longstanding problem of level-spacing distribution in mixed systems. PMID- 14754051 TI - Magnetoelectric Jones dichroism in atoms. AB - The authors suggest that atomic experiments measuring the interference between magnetic-dipole and electric-field-induced electric-dipole transition amplitudes provide a valuable system to study magnetoelectric Jones effects. PMID- 14754052 TI - Optical analogue of electronic Bloch oscillations. AB - We report on the observation of Bloch oscillations in light transport through periodic dielectric systems. By introducing a linear refractive index gradient along the propagation direction the optical equivalent of a Wannier-Stark ladder was obtained. Bloch oscillations were observed as time-resolved oscillations in transmission, in direct analogy to electronic Bloch oscillations in conducting crystals where the Wannier-Stark ladder is obtained via an external electric field. The observed oscillatory behavior is in excellent agreement with transfer matrix calculations. PMID- 14754053 TI - Molding and stretched evolution of optical solitons in cumulative nonlinearities. AB - The observation of initial time dynamics of self-trapping in photorefractive media indicates that optical spatial solitons supported by intense cumulative nonlinearities manifest temporally nonlocal signatures in the form of stretched exponential behavior. This general result, supported also by numerical predictions, is triggered by wave shaping in a time-constant buildup map, a consequence of the spatially resolved inertial response intrinsic to the geometrical transition from a diffracting to a self-focused beam, inherent to soliton appearance. PMID- 14754054 TI - Superstable granular heap in a thin channel. AB - We observed experimentally a new regime for granular flows in an inclined channel with a flow-rate-controlled system. For high flow rates, the flow occurs atop a static granular heap whose angle is considerably higher than those usually exhibited by granular heaps. The properties of such superstable heaps (SSH) are drastically affected by a change in the channel width W. This indicates that the unusual stability of these heaps can be accounted for by the flowing layer and its friction on the sidewalls. A simple depth-averaged model, assuming Coulomb friction, shows that the SSH angle scales as h/W (W being the channel width), and that grain size plays no part. PMID- 14754043 TI - Three-pion Hanbury Brown-Twiss correlations in relativistic heavy-ion collisions from the STAR experiment. AB - Data from the first physics run at the Relativistic Heavy-Ion Collider at Brookhaven National Laboratory, Au+Au collisions at sqrt[s(NN)]=130 GeV, have been analyzed by the STAR Collaboration using three-pion correlations with charged pions to study whether pions are emitted independently at freeze-out. We have made a high-statistics measurement of the three-pion correlation function and calculated the normalized three-particle correlator to obtain a quantitative measurement of the degree of chaoticity of the pion source. It is found that the degree of chaoticity seems to increase with increasing particle multiplicity. PMID- 14754055 TI - Speed of sound in periodic elastic composites. AB - We consider the low-frequency limit (homogenization) for propagation of sound waves in periodic elastic medium (phononic crystals). Exact analytical formulas for the speed of sound propagating in a three-dimensional periodic arrangement of liquid and gas or in a two-dimensional arrangement of solids are derived. We apply our formulas to the well-known phenomenon of the drop of the speed of sound in mixtures. For air bubbles in water we obtain a perfect agreement with the recent results of coherent potential approximation obtained by Phys. Rev. Lett. 84, 6050 (2000)] if the filling of air bubbles is far from close packing. When air spheres almost touch each other, the approximation gives 10 times lower speed of sound than the exact theory does. PMID- 14754056 TI - Distribution of injected power fluctuations in electroconvection. AB - We report on the distribution spectra of the fluctations in the amount of power injected into a liquid crystal undergoing electroconvective flow. The probability distribution functions (PDFs) of the fluc-tuations as well as the magnitude of the fluctuations have been determined in a wide range of imposed stress both for unconfined and confined flow geometries. These spectra are compared to those found in other systems held far from equilibrium, and find that in certain conditions we obtain the universal PDF form reported by Phys. Rev. Lett. 84, 3744 (2000)]. Moreover, the PDF approaches this universal form via an interesting mechanism whereby the distribution's negative tail evolves towards form in a different manner than the positive tail. PMID- 14754044 TI - Pion-Kaon correlations in central Au+Au collisions at square root [sNN] = 130 GeV. AB - Pion-kaon correlation functions are constructed from central Au+Au STAR data taken at sqrt[s(NN)]=130 GeV by the STAR detector at the Relativistic Heavy Ion Collider (RHIC). The results suggest that pions and kaons are not emitted at the same average space-time point. Space-momentum correlations, i.e., transverse flow, lead to a space-time emission asymmetry of pions and kaons that is consistent with the data. This result provides new independent evidence that the system created at RHIC undergoes a collective transverse expansion. PMID- 14754057 TI - Fluctuations do matter: large noise-enhanced halos in charged-particle beams. AB - The formation of beam halos has customarily been described in terms of a particle core model in which the space-charge field of the oscillating core drives particles to large amplitudes. This model involves parametric resonance and predicts a hard upper bound to the orbital amplitude of the halo particles. We show that the presence of colored noise due to space-charge fluctuations and/or machine imperfections can eject particles to much larger amplitudes than would be inferred from parametric resonance alone. PMID- 14754058 TI - New benchmarks from tokamak experiments for theoretical calculations of the dielectronic satellite spectra of heliumlike ions. AB - Dielectronic satellite spectra of heliumlike argon, recorded with a high resolution x-ray crystal spectrometer at the National Spherical Torus Experiment, were found to be inconsistent with existing predictions resulting in unacceptable values for the power balance and suggesting the unlikely existence of non Maxwellian electron energy distributions. These problems were resolved with calculations from a new atomic code. It is now possible to perform reliable electron-temperature measurements and to eliminate the uncertainties associated with determinations of non-Maxwellian distributions. PMID- 14754059 TI - Self-compression of laser pulses in plasma. AB - We study self-compression of weakly relativistically intense laser pulses in subcritical plasmas using one- (1D) and three-dimensional (3D) direct particle-in cell (PIC) simulations. The self-compression works in the density window from 1/4 critical to slightly below critical density, where the Raman instability is prohibited. An analytical model is developed to describe the self-compression. The model admits pulsing Gaussian solutions and a long-lived running soliton solution. The 1D PIC results agree well with the analytical model, and compressions by an order of magnitude are observed. In the 3D geometry, the longitudinal self-compression competes with the transverse self focusing/filamentation. To damp the filamentation we use a periodic plasma-vacuum structure. The 3D PIC simulations suggest that a 30 fs long laser pulse is efficiently compressed to 5 fs. PMID- 14754060 TI - Direct excitation of high-amplitude chirped bucket-BGK modes. AB - For the first time, high amplitude (Deltan/n approximately 40%), high Q (up to 100 000) Bernstein, Greene, and Kruskal modes have been controllably excited in a plasma. The modes are created by sweeping an excitation voltage downwards in frequency, thereby dragging a phase space "bucket" of low density into the bulk of the plasma velocity distribution. The modes have no linear limit and differ markedly from plasma waves and Trivelpiece-Gould modes. PMID- 14754061 TI - Density-transition scale at quasiperpendicular collisionless shocks. AB - Measurements of a spacecraft floating potential, on the four Cluster spacecraft, are used as a proxy for electron plasma density to study, for the first time, the macroscopic density transition scale at 98 crossings of the quasiperpendicular terrestrial bow shock. A timing analysis gives shock speeds and normals; the shock speed is used to convert the temporal measurement to a spatial one. A hyperbolic tangent function is fitted to each density transition, which captures the main shock transition, but not overshoot or undershoot nor foot features. We find that, at a low Mach number M, the density transition is consistent with both ion inertial scales c/omega(pi) and convected gyroradii v(sh,n)/Omega(ci,2), while at M>/=4-5 only the convected gyroradius is the preferred scale for the shock density transition and takes the value L approximately 0.4v(sh,n)/Omega(ci,2). PMID- 14754062 TI - In situ synchrotron study of phase transformation behaviors in bulk metallic glass by simultaneous diffraction and small angle scattering. AB - We have used a new approach involving simultaneous diffraction and small angle scattering to study the amorphous-to-crystalline phase transformation in Zr-based bulk metallic glass. In situ, time-resolved data provided the first direct demonstration of a phase separation prior to crystallization. There is evidence that nucleation and growth of the crystalline phase occur in separate stages, with different kinetics. Our data support the view that crystalline nucleation is achieved via short-range diffusion of small atoms (e.g., Ni), whereas the growth is dictated by long-range diffusion. PMID- 14754063 TI - Crossover of microscopic dynamics in metallic supercooled liquid observed by NMR. AB - Nuclear magnetic resonance is used to characterize local atomic motions in the glassy and supercooled liquid states of the bulk metallic glass system Pd43Ni10Cu27P20. The temperature dependence of the Knight shift reveals that certain local atomic motion decreases rapidly below a crossover temperature T(c) down to the glass transition temperature T(g). Above T(c) as well as below T(g) the mean-squared amplitude of local motions depends linearly on the temperature. The observed gradual transition below T(c) is inconsistent with heterogeneity effects. It reveals that qualitative changes of microscopic properties in the supercooled liquid take place at temperatures significantly above T(g). PMID- 14754064 TI - Structure and bonding of dense liquid oxygen from first principles simulations. AB - Using first principles simulations we have investigated the structural and bonding properties of dense fluid oxygen up to 180 GPa. We have found that band gap closure occurs in the molecular liquid, with a "slow" transition from a semiconducting to a poor metallic state occurring over a wide pressure range. At approximately 80 GPa, molecular dissociation is observed in the metallic fluid. Spin fluctuations play a key role in determining the electronic structure of the low pressure fluid, while they are suppressed at high pressure. PMID- 14754065 TI - In situ transmission-electron-microscopy investigation of melting in submicron Al Si alloy particles under electron-beam irradiation. AB - In situ heating and electron-beam irradiation in the transmission electron microscope were performed to study melting of submicron Al-11.6 at. % Si particles supported on a C thin film. It was found that electron irradiation could be used to melt the particles, even when the hot-stage sample holder was kept at a much lower temperature ( approximately 125 degrees C) than the initial melting point of the particles. Comparison between the experimentally observed melting behavior and analytical calculations indicate that melting of the submicron Al-Si particles under electron-beam irradiation is caused by a temperature rise due to electron thermal spikes in the particles and poor thermal conduction away from the particles. These results have important implications in transmission electron microscopy studies of nanoparticles supported on thin films or poorly conducting substrates. PMID- 14754066 TI - Semisharp phase field method for quantitative phase change simulations. AB - The standard phase field model for simulation of phase change requires an asymptotic analysis in a vanishing interface width, in order to connect the model parameters to the sharp interface parameters, which has hampered the quantitative usefulness of the method. In this Letter the method is simplified to the point that the relevant reduced problem can be solved analytically, allowing the sharp and phase field parameters to be identified, in principle, without restrictions on the model parameters. The scheme is tested for standard cases of two dimensional solidification, showing excellent agreement with sharp interface kinetics. PMID- 14754067 TI - Behavior of complex knots in single DNA molecules. AB - We used optical tweezers to tie individual DNA molecules in knots. Although these knots become highly localized under tension, they remain surprisingly mobile and undergo thermal diffusion with classical random walk statistics. The diffusion constants of knots with different complexities correlate with theoretical calculations of knot sizes. We show that this correlation can be explained by a simple hydrodynamical model of "self-reptation" of the knot along a polymer. PMID- 14754068 TI - Heat-induced transformation of nanodiamond into a tube-shaped fullerene: a molecular dynamics simulation. AB - Heat-induced structural transformation in nanodiamond of diameter approximately 1.4 nm is investigated by tight-binding molecular dynamics simulations using the environment-dependent tight-binding carbon potential. The nanodiamond is found to transform into a tube-shaped fullerene via annealing. Three interesting mechanisms for promoting inner carbon atoms of the nanodiamond into the surface carbon atoms of the tubular structure are observed. The "flow-out" mechanism prevails at temperatures lower than 2500 K and the "direct adsorption" and "push out" mechanisms are observed at higher temperatures. PMID- 14754069 TI - Correct height measurement in noncontact atomic force microscopy. AB - We demonstrate that topography measurements by noncontact atomic force microscopy are subject to residual electrostatic forces. On highly oriented pyrolitic graphite (HOPG) with a submonolayer coverage of C60, we monitor the step height from C60 to HOPG as a function of dc bias between tip and sample. Because of the different contact potential of C60 and HOPG ( approximately 50 mV), the step height is strongly dependent on the dc bias. The presented results and additional simulations demonstrate clearly that for correct height measurements it is mandatory to use a Kelvin probe force microscopy method with active compensation of electrostatic forces. PMID- 14754070 TI - Catalytic role of metal oxides in gold-based catalysts: a first principles study of CO oxidation on TiO2 supported Au. AB - CO oxidation on TiO2 supported Au has been studied using density functional theory calculations. Important catalytic roles of the oxide have been identified: (i) CO oxidation occurs at the interface between Au and the oxide with a very small barrier; and (ii) O2 adsorption at the interface is the key step in the reaction. The physical origin of the oxide promotion effect has been further investigated: The oxide enhances electron transfer from the Au to the antibonding states of O2, giving rise to (i) strong ionic bonding between the adsorbed O2, Au, and the Ti cation; and (ii) a significant activation of O2 towards CO oxidation. PMID- 14754071 TI - Competition between diffusion and fragmentation: an important evolutionary process of nature. AB - We investigate systems of nature where the common physical processes diffusion and fragmentation compete. We derive a rate equation for the size distribution of fragments. The equation leads to a third order differential equation which we solve exactly in terms of Bessel functions. The stationary state is a universal Bessel distribution described by one parameter, which fits perfectly experimental data from two very different systems of nature, namely, the distribution of ice crystal sizes from the Greenland ice sheet and the length distribution of alpha helices in proteins. PMID- 14754072 TI - Enhanced ionic conductivity of polymer electrolytes containing nanocomposite SiO2 particles. AB - We report a new kind of polyethlene oxide (PEO)-Li based composite polymer electrolyte containing active nanocomposite particles with ethylene carbonate (EC)/propylene carbonate (PC) plasticizers embedded in mesoporous SiO2 (EC/PC SiO2). A very large enhancement of the ionic conductivity was observed for the PEO-Li/(EC/PC-SiO2) electrolytes. This could be attributed mainly to conducting EC/PC nanochannels in the active nanocomposite particles. This unique mechanism is absent in currently known nanocomposite polymer electrolytes with inert oxide nanoparticles. Both high ionic conductivity and excellent stability of such new nanocomposite polymer electrolytes potentially makes it one of the most promising electrolyte materials. PMID- 14754073 TI - Anomalous grain boundary physics in polycrystalline CuInSe2: the existence of a hole barrier. AB - First-principles modeling of grain boundaries (GB) in CuInSe2 semiconductors reveals that an energetic barrier exists for holes arriving from the grain interior (GI) to the GB. Consequently, the absence of holes inside the GB prevents GB electrons from recombining. At the same time, the GI is purer in polymaterials than in single crystals, since impurities segregated to the GBs. This explains the puzzle of the superiority of polycrystalline CuInSe2 solar cells over their crystalline counterpart. We identify a simple and universal mechanism for the barrier, arising from reduced p-d repulsion due to Cu-vacancy surface reconstruction. This discovery opens up possibilities for the future design of superior polycrystalline devices. PMID- 14754074 TI - Oscillatory nonlinear conductance of an interacting quantum wire with an impurity. AB - The nonlinear conductance of a one-dimensional quantum wire adiabatically coupled to Fermi liquid electron reservoirs is determined in the presence of an impurity. We show that electron-electron interaction in connection with the finite length of the wire leads to characteristic oscillations in the current as a function of the applied voltage. PMID- 14754075 TI - New mixed alkali effect in the ac conductivity of ion-conducting glasses. AB - We have determined the first wide-range conductivity spectra of a mixed alkali glass system, extending from 10(-3) Hz to 1.3 THz. We report on a new mixed alkali effect in the ac conductivity that is evident even if the low-frequency contributions (linked to hopping motions of the mobile cations) are removed from the experimental spectra. From our results we conclude that the nonhopping contributions to the ac conductivity involve both the mobile ions and the glassy network. PMID- 14754076 TI - Fermi-edge singularity in a nonequilibrium system. AB - We report exact nonperturbative results for the Fermi-edge singularity in the absorption spectrum of an out-of-equilibrium tunnel junction. We consider two metals with chemical potential difference V separated by a tunneling barrier containing a defect, which exists in one of two states. When it is in its excited state, tunneling through the otherwise impermeable barrier is possible. Our nonperturbative solution of this nonequilibrium many-body problem shows that, as well as extending below the equilibrium threshold, the line shape depends on the difference in the phase of the reflection amplitudes on the two sides of the barrier. These results have a surprisingly simple interpretation in terms of known results for the equilibrium case but with (in general complex-valued) combinations of elements of the scattering matrix replacing the equilibrium phase shifts. PMID- 14754077 TI - Coulomb blockade and kondo effect in a quantum Hall antidot. AB - We propose a general capacitive model for an antidot, which has two localized edge states with different spins in the quantum Hall regime. The capacitive coupling of localized excess charges, which are generated around the antidot due to magnetic flux quantization, and their effective spin fluctuation can result in Coulomb blockade, h/(2e) Aharonov-Bohm oscillations, and the Kondo effect. The resultant conductance is in qualitative agreement with recent experimental data. PMID- 14754078 TI - Nonequilibrium dynamics of Andreev states in the Kondo regime. AB - The transport properties of a quantum dot coupled to superconducting leads are analyzed. It is shown that the quasiparticle current in the Kondo regime is determined by the nonequilibrium dynamics of subgap states (Andreev states) under an applied voltage. The current at low bias is suppressed exponentially for decreasing Kondo temperature in agreement with recent experiments. We also predict novel interference effects due to multiple Landau-Zener transitions between Andreev states. PMID- 14754079 TI - Josephson behavior of phase-slip lines in wide superconducting strips. AB - Phase-slip lines can be viewed as dynamically created Josephson junctions in a homogeneous superconducting film. In contrast to phase-slip centers, phase-slip lines occur in wide superconducting strips, where the order parameter may vary in two dimensions. We investigated phase-slip lines in two different materials using several methods. We observed Shapiro steps under microwave radiation, which shows that the frequency of the order parameter oscillation is equal to Josephson frequency. A periodic oscillation of a critical current versus the applied magnetic field was found in strips with a hole in the middle. The latter effect provides a clear evidence of macroscopic quantum interference across a phase-slip line. We have used low temperature scanning laser microscopy to visualize the phase-slip lines and to distinguish them from possible local inhomogeneities in the films. PMID- 14754080 TI - Pair-breaking and superconducting state recovery dynamics in MgB2. AB - We present studies of the photoexcited quasiparticle dynamics in MgB2 where, using femtosecond optical techniques, Cooper pair-breaking dynamics (PBD) have been temporally resolved for the first time. The PBD are strongly temperature and photoexcitation intensity dependent. Analysis of the PBD using the Rothwarf Taylor equations suggests that the anomalous PBD arises from the fact that in MgB2 photoexcitation is initially followed by energy relaxation to high frequency phonons instead of, as commonly assumed, e-e thermalization. Furthermore, the bare quasiparticle recombination rate and the probability for pair breaking by phonons have been determined. PMID- 14754081 TI - Dynamical Coulomb blockade and spin-entangled electrons. AB - We consider the creation of mobile and nonlocal spin-entangled electrons from tunneling of a BCS-superconductor (SC) to two normal leads of finite resistivity. The resulting dynamical Coulomb blockade effect, which we describe phenomenologically in terms of an electromagnetic environment, is shown to be enhanced for tunneling of two electrons from a Cooper pair into the same lead compared to the desired pair-split process where each electron enters a different lead. Conversely, this latter process is suppressed by a finite separation between the tunneling points on the SC. PMID- 14754082 TI - New magnetic order in buried native iron oxide layers. AB - The properties of a magnetically ordered buried Fe oxide layer are presented. This oxide has a room-temperature magnetization exceeding that of Fe3O4 by 42% and of gamma-Fe2O3 by 89%. The oxide consists of a component (70%) with a net moment of 2.0 micro(B)/Fe ion, while the remaining spins yield no net moment. The oxide magnetization is stabilized in part by the proximate Fe metal. PMID- 14754083 TI - Direct imaging and determination of the uncompensated spin density in exchange biased CoO/(CoPt) multilayers. AB - Magnetic force microscopy (MFM) measurements were performed on an exchange-biased CoO/(CoPt) multilayer sample at 7.5 K. Applying an external magnetic field of up to 7 T saturates the ferromagnetic layer and the remaining uncompensated antiferromagnetic spins at the antiferromagnet-ferromagnet interfaces are imaged with high lateral resolution. The coupling between the uncompensated spins and the spins in the ferromagnet are found to be antiferromagnetic. Quantitative analysis of the MFM images revealed that 7% of the spins at the interface are uncompensated and contribute to the exchange biasing. PMID- 14754084 TI - Spin dependence in the survival probability of metastable He (2(3)S) atoms during the scattering from ferromagnetic surfaces. AB - The use of a spin-polarized metastable helium (He(*)) atom beam has demonstrated that the survival probability (SP) of He(*) during the scattering from ferromagnetic surfaces depends on the electron spin of He(*). The spin asymmetry in the SP of He(*) scattered from a clean Fe/Cu(100) surface changed its polarity with H2O adsorption. We argue that the reversal is caused by the change in the spin polarization of the surface electronic states at around the Fermi level. PMID- 14754085 TI - Range of magnetic correlations in nanocrystalline soft magnets. AB - We have obtained the magnetic field dependence of static ferromagnetic correlations in nanocrystalline electrodeposited Co and Ni by means of the correlation function of the spin misalignment, determined from small-angle neutron scattering data. The approach yields a correlation length l(C), which is a measure for the spatial extent of inhomogeneities in the magnetization distribution. The correlation length depends strongly on the applied magnetic field with values ranging from 94 nm in nanocrystalline Co at low fields to about 15 nm at saturation. The results for l(C) indicate that in Co the main source of nonuniformity in the spin system is the anisotropy field of each individual crystallite, whereas in nanocrystalline Ni the main sources of spin disorder originate from twin faults or from the defect cores of grain boundaries. PMID- 14754086 TI - Nominal thermodynamic temperature in nonequilibrium kinetic Ising models. AB - We show that a nominal temperature can be consistently and uniquely defined everywhere in the phase diagram of large classes of nonequilibrium kinetic Ising spin models. In addition, we confirm that, at critical points, the large-time "fluctuation-dissipation ratio" X( infinity ) is a universal amplitude ratio, and find in particular X( infinity ) approximately 0.33(1) and X( infinity )=1 / 2 for the magnetization in, respectively, the two-dimensional Ising and voter universality classes. PMID- 14754087 TI - Exciton dephasing in quantum dot molecules. AB - We have measured the exciton dephasing time in InAs/GaAs quantum dot molecules having different interdot barrier thicknesses in the temperature range from 5 to 60 K, using a highly sensitive four-wave mixing heterodyne technique. At 5 K dephasing times of several hundred picoseconds are found. Moreover, a systematic dependence of the dephasing dynamics on the barrier thickness is observed. These results show how the quantum-mechanical coupling of the electronic wave functions in the molecules affects both the exciton radiative lifetime and the exciton acoustic phonon interaction. PMID- 14754088 TI - Infinite-order excitonic BLOCH equations for asymmetric nanostructures. AB - We present a new exciton-based formalism for calculating the coherent response of asymmetric semiconductor multiple quantum well structures to ultrashort optical pulses valid to infinite order in the optical field and including the self generated intraband fields. We use these equations to calculate and explain the oscillations with time delay of peaks in the spectrally resolved degenerate four wave mixing signals from biased semiconductor superlattices, obtaining good agreement with experiment. PMID- 14754089 TI - Linewidth-limited energy transfer in single conjugated polymer molecules. AB - Using low temperature single molecule spectroscopy on rigid-rod conjugated polymers we are able to identify homogeneously broadened, strongly polarized emission from individual chromophore units on a single chain. Gated fluorescence spectroscopy allows real time imaging of intramolecular energy transfer as the chain behaves as a series of weakly interacting chromophores. Energy transfer is controlled by the chromophoric spectral linewidth, which depends on temperature. Linewidths exceeding intramolecular disorder lead to incoherent chromophore coupling and collective fluorescence phenomena. PMID- 14754090 TI - Dielectric discontinuity at interfaces in the atomic-scale limit: permittivity of ultrathin oxide films on silicon. AB - Using a density-functional approach, we study the dielectric permittivity across interfaces at the atomic scale. Focusing on the static and high-frequency permittivities of SiO2 films on silicon, for oxide thicknesses from 12 A down to the atomic scale, we find a departure from bulk values in accord with experiment. A classical three-layer model accounts for the calculated permittivities and is supported by the microscopic polarization profile across the interface. The local screening varies on length scales corresponding to first-neighbor distances, indicating that the dielectric transition is governed by the chemical grading. Silicon-induced gap states are shown to play a minor role. PMID- 14754091 TI - Spontaneous oscillations of dressed neurons: a new mechanism for epilepsy? AB - Most modeling studies of neurons and neuronal networks are based on the assumption that the neurons are isolated from their normal environment. Based on recent experimental data we put forward a model for neurons that incorporates the influence of the surrounding glia (dressed neurons). We predict seizurelike spontaneous oscillations in the absence of stimuli for strong coupling between neurons and astrocytes. Consistent with our predictions, a signature of this enhanced crosstalk, over expression of glutamate receptors in astrocytes, has been observed specifically in epileptic tissue. PMID- 14754092 TI - Evidence of a higher-order singularity in dense short-ranged attractive colloids. AB - We study a model in which particles interact through a hard-core repulsion complemented by a short-ranged attractive potential of the kind found in colloidal suspensions. Combining theoretical and numerical work we locate the line of higher-order glass-transition singularities and its end point-named A4-on the fluid-glass line. Close to the A4 point, we detect logarithmic decay of density correlations and a sublinear power-law increase of the mean square displacement, for time intervals up to 4 orders of magnitude. We establish the presence of the A4 singularity by studying how the range of the potential affects the time window where anomalous dynamics is observed. PMID- 14754093 TI - Relaxation dynamics of langmuir polymer films: a power-law analysis. AB - We report an experimental study on the mechanical relaxation of Langmuir films of a flexible polymer: polyvinylacetate, a good example of a two-dimensional polymeric system at good-solvent conditions. This study allows us to explore the dependence of the relaxation times and the surface viscosity on the concentration. In the semidilute regime, both dynamical properties follow well defined power laws, which are in quantitative agreement with 2D reptation. PMID- 14754094 TI - Comment on "Nonclassical paths in the recurrence spectrum of diamagnetic atoms". PMID- 14754096 TI - Comment on "Algebraic Fermi liquid from phase fluctuations: 'topological' fermions, vortex 'berryons,' and QED3 theory of cuprate superconductor". PMID- 14754098 TI - Observation of Bose-Einstein condensation of molecules. AB - We have observed Bose-Einstein condensation of molecules. When a spin mixture of fermionic 6Li atoms was evaporatively cooled in an optical dipole trap near a Feshbach resonance, the atomic gas was converted into 6Li2 molecules. Below 600 nK, a Bose-Einstein condensate of up to 900 000 molecules was identified by the sudden onset of a bimodal density distribution. This condensate realizes the limit of tightly bound fermion pairs in the crossover between BCS superfluidity and Bose-Einstein condensation. PMID- 14754099 TI - Exciting collective oscillations in a trapped 1D gas. AB - We report on the realization of a trapped one-dimensional Bose gas and its characterization by means of measuring its lowest lying collective excitations. The quantum degenerate Bose gas is prepared in a 2D optical lattice, and we find the ratio of the frequencies of the lowest compressional (breathing) mode and the dipole mode to be (omega(B)/omega(D))(2) approximately 3.1, in accordance with the Lieb-Liniger and mean-field theory. For a thermal gas we measure (omega(B)/omega(D))(2) approximately 4. By heating the quantum degenerate gas, we have studied the transition between the two regimes. For the lowest number of particles attainable in the experiment the kinetic energy of the system is similar to the interaction energy, and we enter the strongly interacting regime. PMID- 14754100 TI - Quantitative test of thermal field theory for Bose-Einstein condensates. AB - We present numerical results from a second-order quantum field theory of Bose Einstein condensates applied to the 1997 JILA experiment [Phys. Rev. Lett. 78, 764 (1997)]]. Good agreement is found for the energies and decay rates for both the lowest-energy m=2 and m=0 modes. The anomalous behavior of the m=0 mode is due to experimental perturbation of the noncondensate. The theory is gapless and includes the coupled dynamics of the condensate and thermal cloud, the anomalous pair average, and all relevant finite size effects. PMID- 14754101 TI - Spectroscopic insensitivity to cold collisions in a two-state mixture of fermions. AB - We have experimentally demonstrated the absence of spectroscopic resonance shifts in a mixture of two interacting Fermi gases. This result is linked to observations in an ultracold gas of thermal bosons. There, the measured resonance shift due to interstate collisions is independent of the coherence in the system, and twice that expected from the equilibrium energy splitting between the two internal states in a fully decohered cloud. We give a simple theoretical explanation of these observations, which elucidates the effect of coherent radiation on an incoherent mixture of atoms. PMID- 14754102 TI - Quantum Kalman filtering and the Heisenberg limit in atomic magnetometry. AB - The shot-noise detection limit in current high-precision magnetometry [Nature (London) 422, 596 (2003)] is a manifestation of quantum fluctuations that scale as 1/sqrt[N] in an ensemble of N atoms. Here, we develop a procedure that combines continuous measurement and quantum Kalman filtering [Rep. Math. Phys. 43, 405 (1999)]] to surpass this conventional limit by exploiting conditional spin squeezing to achieve 1/N field sensitivity. Our analysis demonstrates the importance of optimal estimation for high bandwidth precision magnetometry at the Heisenberg limit and also identifies an approximate estimator based on linear regression. PMID- 14754103 TI - Are cluster magnetic fields primordial? AB - We present results of a detailed and fully nonlinear numerical and analytical investigation of magnetic field evolution from the very earliest cosmic epochs to the present. We find that, under reasonable assumptions concerning the efficiency of a putative magnetogenesis era during cosmic phase transitions, surprisingly strong magnetic fields 10(-13)-10(-11) G on comparatively small scales 100 pc-10 kpc may survive to the present. Building on prior numerical work on the evolution of magnetic fields during the course of gravitational collapse of a cluster, which indicates that precollapse fields of approximately 4 x 10(-12) G extant on small scales may suffice to produce clusters with acceptable Faraday rotation measures, we argue that it seems possible for cluster magnetic fields to be entirely of primordial origin. PMID- 14754104 TI - Radiative electroweak symmetry breaking revisited. AB - In the absence of a tree-level scalar-field mass, renormalization-group methods permit the explicit summation of leading-logarithm contributions to all orders of the perturbative series within the effective potential for SU(2)xU(1) electroweak symmetry. This improvement of the effective potential function is seen to reduce residual dependence on the renormalization mass scale. The all-orders summation of leading-logarithm terms involving the dominant three couplings contributing to radiative corrections is suggestive of a potential characterized by a plausible Higgs boson mass of 216 GeV. However, the tree potential's local minimum at phi=0 is restored if QCD is sufficiently strong. PMID- 14754105 TI - Planar amplitudes in maximally supersymmetric Yang-Mills theory. AB - The collinear factorization properties of two-loop scattering amplitudes in dimensionally regulated N=4 super-Yang-Mills theory suggest that, in the planar ('t Hooft) limit, higher-loop contributions can be expressed entirely in terms of one-loop amplitudes. We demonstrate this relation explicitly for the two-loop four-point amplitude and, based on the collinear limits, conjecture an analogous relation for n-point amplitudes. The simplicity of the relation is consistent with intuition based on the anti-de Sitter/conformal field theory correspondence that the form of the large-N(c) L-loop amplitudes should be simple enough to allow a resummation to all orders. PMID- 14754106 TI - Leptogenesis from supersymmetry breaking. AB - We show that soft supersymmetry breaking terms involving the heavy sneutrinos can lead to sneutrino-antisneutrino mixing and to new sources of CP violation, which are present even if a single generation is considered. These terms are naturally present in supersymmetric versions of leptogenesis scenarios, and they induce indirect CP violation in the decays of the heavy sneutrinos, eventually generating a baryon asymmetry. This new contribution can be comparable to or even dominate over the asymmetry produced in traditional leptogenesis scenarios. PMID- 14754108 TI - Determination of the 8B neutrino spectrum. AB - We have measured the total energy of the alpha particles following the beta decay of 8B by implanting 8B into a planar silicon surface barrier detector. Calibration was performed using alpha particles following the beta decay of 20Na, similarly implanted. The alpha spectrum is used to infer the 8B neutrino spectrum which is an important input in the interpretation of experiments that detect energetic neutrinos from the Sun. The alpha spectrum reported here is in disagreement with the previous best measurement which used two detectors in coincidence. PMID- 14754109 TI - Quantum coherence in the time-resolved Auger measurement. AB - We present a quantum mechanical model of the attosecond-XUV (extreme ultraviolet) pump and laser probe measurement of an Auger decay [Drescher et al., Nature (London) 419, 803 (2002)]] and investigate effects of quantum coherence. The time dependent Schrodinger equation is solved by numerical integration and in analytic form. We explain the transition from a quasiclassical energy shift of the spectrum to the formation of sidebands and the enhancement of high- and low energy tails of the Auger spectrum due to quantum coherence between photoionization and Auger decay. PMID- 14754110 TI - Interaction-induced decoherence of atomic BLOCH oscillations. AB - We show that the energy spectrum of the Bose-Hubbard model amended by a static field exhibits Wigner-Dyson level statistics. In itself a characteristic signature of quantum chaos, this induces the irreversible decay of Bloch oscillations of cold, interacting atoms loaded into an optical lattice, and provides a Hamiltonian model for interaction-induced decoherence. PMID- 14754111 TI - Chaotic filtering of moving atoms in pulsed optical lattices by control of dynamical localization. AB - We propose a mechanism for a velocity-selective device which would allow packets of cold atoms traveling in one direction through a pulsed optical lattice to pass undisturbed, while dispersing atoms traveling in the opposite direction. The mechanism is generic and straightforward: for a simple quantum kicked rotor pulsed with unequal periods, the quantum suppression of momentum diffusion (dynamical localization) yields momentum localization lengths L which are no longer isotropic, as in the standard case, but vary smoothly and controllably with initial momentum. PMID- 14754112 TI - Measurement of the phase of few-cycle laser pulses. AB - For the shortest pulses generated to date, the amplitude of the electromagnetic wave changes almost as rapidly as the field oscillates. The temporal variation of the field, which directly governs strong-field interactions, therefore depends on whether the maximum of the pulse amplitude coincides with that of the wave cycle or not, i.e., on the phase of the field with respect to the pulse envelope. It is demonstrated that the direction of electron emission from photoionized atoms can be controlled by varying the phase of the field, providing for the first time a tool for its accurate determination. Directing fast electron emission to the right or to the left with the light phase constitutes a new kind of coherent control. PMID- 14754107 TI - Observation of an exotic S = +1 baryon in exclusive photoproduction from the deuteron. AB - In an exclusive measurement of the reaction gammad-->K(+)K(-)pn, a narrow peak that can be attributed to an exotic baryon with strangeness S=+1 is seen in the K(+)n invariant mass spectrum. The peak is at 1.542+/-0.005 GeV/c(2) with a measured width of 0.021 GeV/c(2) FWHM, which is largely determined by experimental mass resolution. The statistical significance of the peak is (5.2+/ 0.6)sigma. The mass and width of the observed peak are consistent with recent reports of a narrow S=+1 baryon by other experimental groups. PMID- 14754113 TI - Probing scattering wave functions close to the nucleus. AB - Recently, three-dimensional imaging of the ejected electrons following 100 MeV/amu C6+ single ionization of helium led to the observation of a new structure not predicted by theory [Nature (London) 422, 48 (2003)]]. Instead of the usual "recoil lobe" centered on the momentum-transfer axis, a ring-shaped structure centered on the beam axis was observed. New measurements at 2 MeV/amu exhibit a similar structure, which is now predicted by theory. We argue that the same theory failed at 100 MeV/amu because the faster projectiles probe distances much closer to the nucleus, where our multiple-scattering model is expected to break down. PMID- 14754114 TI - Integral representation for the electron-atom ionization amplitude which is free of ambiguity and divergence problems. AB - It is shown that existing problems with the formal theory of ionization can be effectively resolved. An integral representation for the ionization amplitude free of ambiguity and divergence problems is given. Moreover, the ionization amplitude in the new formulation is shown directly to have an ideal form for practical calculations. PMID- 14754115 TI - Lamb shift of laser-dressed atomic states. AB - We discuss radiative corrections to an atomic two-level system subject to an intense driving laser field. It is shown that the Lamb shift of the laser-dressed states, which are the natural state basis of the combined atom-laser system, cannot be explained in terms of the Lamb shift received by the atomic bare states which is usually observed in spectroscopic experiments. In the final part, we propose an experimental scheme to measure these corrections based on the incoherent resonance fluorescence spectrum of the driven atom. PMID- 14754116 TI - Second harmonic generation due to quadrupole interaction in a photonic crystal slab: angle dependence and symmetry of the unit cell. AB - We investigate second harmonic generation (SHG) from a photonic crystal slab consisting of centrosymmetric materials. The SHG signal is observed in the transmission direction when the incident laser excites the quasiwaveguide mode. As the SHG frequency approaches the exciton level, the SHG intensity increases resonantly. When the incident angle is exactly 0, the SHG signal vanishes even if the transmission dip is excited. This fact is readily explained by a quadrupole theory based on the Lorentz oscillator model, where the source of the nonlinearity is the Lorentz force. When the unit cell in the photonic crystal lacks inversion symmetry, the SHG signal is expected even for the normal incidence. It is experimentally demonstrated for a square array of triangular semiconductor slabs. PMID- 14754117 TI - Resonant behavior of dielectric objects (electrostatic resonances). AB - Resonant behavior of dielectric objects occurs at certain frequencies for which the object permittivity is negative and the free-space wavelength is large in comparison with the object dimensions. Unique physical features of these resonances are studied and a novel technique for the calculation of resonance values of permittivity, and hence resonance frequencies, is proposed. Scale invariance of resonance frequencies, unusually strong orthogonality properties of resonance modes, and a two-dimensional phenomenon of "twin" spectra are reported. The paper concludes with brief discussions of optical controllability of these resonances in semiconductor nanoparticles and a plausible, electrostatic resonance based, mechanism for nucleation and formation of ball lightning. PMID- 14754118 TI - New method for computing finite-time Lyapunov exponents. AB - We present a novel method for computing finite-time Lyapunov exponents and vectors, via generalizing a correction given by Goldhirsch, Sulem, and Orszag [Physica (Amsterdam) 27D, 311 (1987)]] into higher-order corrections. This method is a generalized LR method, which is, in contrast to the existing methods, applicable to multidimensional systems with degenerate spectra. The efficiency and accuracy is demonstrated by applying it to multidimensional dynamical systems. Without these corrections, we could not accurately detect, as an example, the coexistence of qualitatively different Lyapunov instabilities along a trajectory for a multidimensional oscillator system. PMID- 14754119 TI - Quantum necking in stressed metallic nanowires. AB - When a macroscopic metallic wire is subject to tensile stress, it necks down smoothly as it elongates. We show that nanowires with radii comparable to the Fermi wavelength display remarkably different behavior. Using concepts from fluid dynamics, a partial differential equation for nanowire shape evolution is derived from a semiclassical energy functional that includes electron-shell effects. A rich dynamics involving movement and interaction of kinks connecting locally stable radii is found, and a new class of universal equilibrium shapes is predicted. PMID- 14754120 TI - Transport and reconnection in tokamak sawteeth. AB - The core of a tokamak discharge often undergoes periodic relaxation oscillations, sawteeth, as the steepening current and temperature profiles are flattened by fast reconnection events. Careful analysis of the electron temperature evolution over this cycle gives an estimate of the energy dissipated in the electrons during reconnection and a measure of the transport characteristic (energy flux versus temperature gradient) over the range of parameters occurring over the remainder of the cycle. The energy dissipated is consistent with estimates of the loss of poloidal magnetic energy. The transport characteristics exhibit a wide range of behaviors. PMID- 14754121 TI - Spatial uniformity of laser-accelerated ultrahigh-current MeV electron propagation in metals and insulators. AB - The evolution of laser-generated MeV, MA electron beams propagating through conductors and insulators has been studied by comparing measurement and modeling of the distribution of MeV protons that are sheath accelerated by the propagated electrons. We find that electron flow through metals is uniform and can be laser imprinted, whereas propagation through insulators induces spatial disruption of the fast electrons. Agreement is found with material dependent modeling. PMID- 14754122 TI - Transverse optical mode in a one-dimensional Yukawa chain. AB - A transverse optical mode was observed in a one-dimensional Yukawa chain. Charged particles, suspended in a strongly coupled dusty plasma, were arranged in a 1D periodic structure. Particle displacement in the direction perpendicular to the chain was restored by the confining potential. The dispersion relation of phonons was measured, verifying that the optical mode has negative dispersion, with phase and group velocities that are oppositely directed. A theoretical dispersion relation is presented and compared to the experiment and a molecular dynamics simulation. PMID- 14754123 TI - Bogoliubov shadow bands in the normal state of superconducting systems with strong pair fluctuations. AB - On the basis of a scenario where electron pairing is induced by resonant two particle scattering (the boson-fermion model), we show how precursors of the superconducting state-in the form of overdamped Bogoliubov modes-emerge in the normal state upon approaching the transition temperature from above. This result is obtained by a renormalization technique based on continuous unitary transformations (the flow equations), projecting out the coherent contributions in the electron spectral function from an incoherent background. PMID- 14754124 TI - Electronic structure of a polydiacetylene nanowire fabricated on highly ordered pyrolytic graphite. AB - A molecular wire candidate, the polydiacetylene chain, fabricated in a substantial support layer of monomers self-assembled on a highly ordered pyrolytic graphite surface, was studied using scanning tunneling microscopy and spectroscopy. The density of states of individual polymers and constituent monomers were observed on the same surface, and then compared with the calculated results. The spectrum delineating the density of states of the polydiacetylene wire clearly reveals the theoretically predicted pi-band and band edge singularities of the one-dimensional polymer. PMID- 14754125 TI - Direct observation of charge transfer in double-perovskite-like RbMn[Fe(CN)6]. AB - The charge density distribution has been determined for a transition metal cyanide, RbMn[Fe(CN)(6)], by means of the maximum entropy-Rietveld method combined with the highly angularly resolved synchrotron radiation x-ray powder diffraction at SPring-8 BL02B2. We directly observed a charge transfer from the Mn site to the Fe site in the low-temperature phase. On the basis of a local density approximation calculation, we discuss the origin for the anisotropic bonding electron distribution around the Mn3+ ion in the low-temperature phase. PMID- 14754126 TI - Transition from Kardar-Parisi-Zhang to tilted interface critical behavior in a solvable asymmetric avalanche model. AB - We use a discrete-time formulation of the asymmetric avalanche process (ASAP) [Phys. Rev. Lett. 87, 084301 (2001)]] of p particles on a finite ring of N sites to obtain an exact expression for the average avalanche size as a function of toppling probabilities and particle density rho=p/N. By mapping the model onto driven interface problems, we find that the ASAP incorporates the annealed Kardar Parizi-Zhang and quenched tilted interface dynamics for rhorho(c), respectively, with rho(c) being the critical density for given toppling probabilities and N--> infinity. We analyze the crossover between two regimes and show which parameters are relevant near the transition point. PMID- 14754127 TI - Atomic structure of antiphase domain boundaries of a thin Al2O3 film on NiAl(110). AB - Line defects of a thin alumina film on NiAl(110) have been studied on the atomic level with scanning tunneling microscopy at 4 K. While boundaries between two reflection domains do not expose a characteristic structure, antiphase domain boundaries are well ordered. The latter boundaries result from the insertion of a row of O atoms, as atomically resolved images of the topmost oxygen layer show. The insertion occurs only in two of the three characteristic directions of the quasihexagonal O lattice. Depending on the direction, either straight or zigzagged boundaries form. An atomic characterization of line defects on the oxide surface is a first step to correlate their topographic structure and chemical activity. PMID- 14754128 TI - Vibrational characterization of the oxidation products on Si(111)-(7 x 7). AB - The oxidation products on Si(111)-(7x7) are investigated at 82 K by means of high resolution electron energy loss spectroscopy. The isotope-labeled vibrational spectra with 16O2, 18O2, and 16O 18O show that, in the initial stage of the oxidation, an O2 molecule dissociates to form a metastable product with an O atom bonding on top of the Si adatom and the other inserted into the backbond. The metastable product is observed as a dark site in the topographic scanning tunneling microscopy (STM) image and can be transformed to a stable product by the STM manipulation. Our results are in good agreement with recent theoretical calculations. PMID- 14754130 TI - Magnetic-field-induced quantum critical point and competing order parameters in URu2Si2. AB - A comprehensive transport study, as a function of temperature and continuous magnetic fields of up to 45 T, reveals that URu2Si2 possesses all the essential hallmarks of quantum criticality at fields around 37+/-1 T. The formation of multiple phases at low temperatures at and around the quantum critical point suggests the existence of competing order parameters. PMID- 14754129 TI - Formation mechanism of H2Ti3O7 nanotubes. AB - Formation mechanism of H2Ti3O7 nanotubes by single-step reaction of crystalline TiO2 and NaOH has been investigated via transmission electron microscopy examinations of series specimens with different reaction times and extensive ab initio calculations. It was found that the growth mechanism includes several steps. Crystalline TiO2 reacts with NaOH, forming a highly disordered phase, which recrystallized into some H2Ti3O7 thin plates. H-deficiency on the top surface leads to an asymmetrical environment for the surface Ti3O2-7 layer. The calculations of the surface tension, elastic strain energy, interlayer coupling energy, and Coulomb force indicated that the asymmetrical environment is the principal driving force of the cleavage of the single sheets of H2Ti3O7 from the plates and the formation of the multiwall spiral nanotubes. PMID- 14754131 TI - Bound excitons in time-dependent density-functional theory: optical and energy loss spectra. AB - A robust and efficient frequency dependent and nonlocal exchange correlation f(xc)(r,r(');omega) is derived by imposing time-dependent density-functional theory (TDDFT) to reproduce the many-body diagrammatic expansion of the Bethe Salpeter polarization function. As an illustration, we compute the optical spectra of LiF, SiO2, and diamond and the finite momentum transfer energy-loss spectrum of LiF. The TDDFT results reproduce extremely well the excitonic effects embodied in the Bethe-Salpeter approach, both for strongly bound and resonant excitons. We provide a working expression for f(xc) that is fast to evaluate and easy to implement. PMID- 14754132 TI - Dynamical mean-field theory of transport of small polarons. AB - We present a unified view of the transport properties of small polarons in the Holstein model at low carrier densities, based on the dynamical mean-field theory. The nonperturbative nature of the approach allows us to study the crossover from classical activated motion at high temperatures to coherent motion at low temperatures. Large quantitative discrepancies from the standard polaronic formulas are found. The scaling properties of the resistivity are analyzed, and a simple interpolation formula is proposed in the nonadiabatic regime. PMID- 14754133 TI - Ta 5d band symmetry of 1T-TaS1.2Se0.8 in the commensurate charge-density-wave phase. AB - We present a detailed angle-resolved photoemission study on the layered transition-metal dichalcogenide 1T-TaS1.2Se0.8 in the commensurate charge-density wave (CDW) phase. A drastic reduction in the spectral weight along the high symmetry line GammaM, particularly around the point M, is observed when s polarized light was used. This implies that the initial state must be symmetric with respect to a mirror plane perpendicular to the line GammaK, which is consistent with conventional band calculations in the absence of the CDW. We conclude that there is only a limited amount of modification of the electronic structure of 1T-TaS1.2Se0.8 in the commensurate CDW phase due to the CDW-related potential. PMID- 14754134 TI - Exact duality relations in correlated electron systems. AB - Using gauge transformations on electron bond operators, we derive exact duality relations between various order parameters for correlated electron systems. Applying these transformations, we find two duality relations in the generalized two-leg Hubbard ladder at arbitrary filling. The relations show that unconventional density-wave orders such as staggered flux or circulating spin current are dual to conventional density-wave orders and there are direct mappings between dual phases. Several exact results on the phase diagram are also concluded. PMID- 14754135 TI - Modulation of thermoelectric power of individual carbon nanotubes. AB - Thermoelectric power (TEP) of individual single walled carbon nanotubes (SWNTs) has been measured at mesoscopic scales using a microfabricated heater and thermometers. Gate electric field dependent TEP modulation has been observed. The measured TEP of SWNTs is well correlated to the electrical conductance across the SWNT according to the Mott formula. Strong modulations of TEP were observed in the single-electron conduction limit. In addition, semiconducting SWNTs exhibit large values of TEP due to the Schottky barriers at SWNT-metal junctions. PMID- 14754136 TI - Image states and excitons at insulator surfaces with negative electron affinity. AB - We discuss electronic excitation processes at two ionic insulator surfaces, LiF(001)-(1x1) and MgO(001)-(1x1), within ab initio many-body perturbation theory. Because of the negative electron affinity of the surfaces, the lowest unoccupied electronic states are image states located in the vacuum outside the surface. Excitations of electrons from the surface layer into these image states are much lower in energy than the bulk excitons. They are responsible for characteristic surface features in the electron energy-loss spectra of LiF(001) and MgO(001). PMID- 14754137 TI - Interplay of Rayleigh and Peierls instabilities in metallic nanowires. AB - A quantum-mechanical stability analysis of metallic nanowires within the free electron model is presented. The stability is determined by an interplay of electron-shell effects, the Rayleigh instability due to surface tension, and the Peierls instability. Although the latter effect limits the maximum length also for wires with "magic radii," it is found that nanowires in the micrometer range can be stable at room temperature. PMID- 14754138 TI - Avoided antiferromagnetic order and quantum critical point in CeCoIn5. AB - We measured the specific heat and resistivity of heavy fermion CeCoIn5 between the superconducting critical field H(c2)=5 T and 9 T, with the field in the [001] direction, and at temperatures down to 50 mK. At 5 T the data show a non-Fermi liquid (NFL) behavior down to the lowest temperatures. At the field above 8 T the data exhibit a crossover from the Fermi liquid to a non-Fermi liquid behavior. We analyzed the scaling properties of the specific heat and compared both the resistivity and the specific heat with the predictions of a spin-fluctuation theory. Our analysis leads us to suggest that the NFL behavior is due to incipient antiferromagnetism (AFM) in CeCoIn5 with the quantum critical point in the vicinity of H(c2). Below H(c2) the AFM phase which competes with the paramagnetic ground state is superseded by the superconducting transition. PMID- 14754139 TI - Thermodynamics and phase diagram of high temperature superconductors. AB - Thermodynamic quantities are derived for superconducting and pseudogap regimes by taking into account both amplitude and phase fluctuations of the pairing field. In the normal (pseudogap) state of the underdoped cuprates, two domains have to be distinguished: near the superconducting region, phase correlations are important up to temperature T(phi). Above T(phi), the pseudogap region is determined only by amplitudes, and phases are uncorrelated. Our calculations show excellent quantitative agreement with specific heat and magnetic susceptibility experiments on cuprates. We find that the mean field temperature T0 has a similar doping dependence as the pseudogap temperature T(*), whereas the pseudogap energy scale is given by the average amplitude above T(c). PMID- 14754140 TI - Electronic state of a CoO2 layer with hexagonal structure: a Kagome lattice structure in a triangular lattice. AB - The electronic state in layered cobalt oxides with a hexagonal structure is examined. We find that the electronic structure reflects the nature of the Kagome lattice hidden in the CoO2 layer which consists of stacked triangular lattices of oxygen ions and of cobalt ions. A fundamental model for the electron system is proposed, and the mechanism of the unique transport and magnetic properties of the cobalt oxides are discussed in light of the model. PMID- 14754141 TI - Quantum dissociation of a vortex-antivortex pair in a long josephson junction. AB - The thermal and the quantum dissociation of a single vortex-antivortex (VAV) pair in an annular Josephson junction is experimentally observed and theoretically analyzed. In our experiments, the VAV pair is confined in a pinning potential controlled by external magnetic field and bias current. The dissociation of the pinned VAV pair manifests itself in a switching of the Josephson junction from the superconducting to the resistive state. The observed temperature and field dependence of the switching current distribution is in agreement with the analysis. The crossover from the thermal to the macroscopic quantum tunneling mechanism of dissociation occurs at a temperature of about 100 mK. We also predict the specific magnetic field dependence of the oscillatory energy levels of the pinned VAV state. PMID- 14754142 TI - Chiral spin liquid wave function and the Lieb-Schultz-Mattis theorem. AB - We study a chiral spin liquid wave function defined as a Gutzwiller projected BCS state with a complex pairing function. After projection, spontaneous dimerization is found for any odd but finite number of chains, thus satisfying the Lieb Schultz-Mattis theorem, whereas for an even number of chains there is no dimerization. The two-dimensional thermodynamic limit is consistently reached for a large number of chains since the dimer order parameter vanishes in this limit. This property clearly supports the possibility of a spin liquid ground state in two dimensions with a gap to all physical excitations and with no broken translation symmetry. PMID- 14754143 TI - Possible spin triplet superconductivity in NaxCoO2.yH2O. AB - Symmetry-based considerations are combined with inputs from available experimental results to make the case that a novel spin-triplet superconductivity triggered by antiferromagnetic fluctuations may be realized in the newly discovered layered cobaltide NaxCoO2.yH(2)O. In the proposed picture, inaccessible via resonating-valence-bond physics extrapolated from half-filling, the pairing process is similar to that advanced for Sr2RuO4, but enjoys a further advantage coming from the hexagonal structure of the Fermi surface which gives a stronger pairing tendency. PMID- 14754144 TI - Photoinduced emission of cooper pairs from superconductors. AB - Under certain conditions specified in this work, a Cooper pair can be emitted from a superconductor upon the absorption of one ultraviolet photon. The spectra of the excited electron pair carry direct information on the energy and angular pair correlation. These statements are concluded from a formal and numerical analysis based on the Bardeen-Cooper-Schrieffer theory for superconductivity. PMID- 14754145 TI - Spin correlations among the charge carriers in an ordered stripe phase. AB - We have observed a diffuse component to the low-energy magnetic excitation spectrum of stripe-ordered La(5/3)Sr(1/3)NiO4 probed by neutron inelastic scattering. The diffuse scattering forms a square pattern with sides parallel and perpendicular to the stripe directions. The signal is dispersive, with a maximum energy of approximately 10 meV. Probed at 2 meV, the scattering decreases in strength with increasing temperature, and is barely visible at 100 K. We argue that the signal originates from dynamic, quasi-one-dimensional, antiferromagnetic correlations among the stripe electrons. PMID- 14754146 TI - Magnetic neutron scattering study of YVO3: evidence for an orbital Peierls state. AB - Neutron spectroscopy has revealed a highly unusual magnetic structure and dynamics in YVO3, an insulating pseudocubic perovskite that undergoes a series of temperature-induced phase transitions between states with different spin and orbital ordering patterns. A good description of the neutron data is obtained by a theoretical analysis of the spin and orbital correlations of a quasi-one dimensional model. This leads to the tentative identification of one of the phases of YVO3 with the "orbital Peierls state," a theoretically proposed many body state comprised of orbital singlet bonds. PMID- 14754147 TI - Dimerization versus orbital-moment ordering in a Mott insulator YVO3. AB - We use exact diagonalization combined with mean-field theory to investigate the phase diagram of the spin-orbital model for cubic vanadates. The spin-orbit coupling competes with Hund's exchange and triggers a novel phase, with the ordering of t(2g) orbital magnetic moments stabilized by the tilting of VO6 octahedra. It explains qualitatively spin canting and reduction of magnetization observed in YVO3. At finite temperature, an orbital instability in the C-type antiferromagnetic phase induces modulation of magnetic exchange constants even in the absence of lattice distortions. The calculated spin structure factor shows a magnon splitting at q-->=(0,0,pi / 2) due to the orbital dimerization. PMID- 14754148 TI - Quantum dynamics of atomic magnets: cotunneling and dipolar-biased tunneling. AB - Multispin tunneling cross relaxations in an ensemble of weakly coupled Ho3+ ions, mediated by weak anisotropic dipolar interactions, can be evidenced by ac susceptibility measurements in a high temperature regime. Based on a four-body representation, including the rare-earth nuclear spin, two-ion tunneling mechanisms can be attributed to both dipolar-biased tunneling and cotunneling processes. The coreversal involving entangled pairs of magnetic moments is discussed with a particular emphasis, giving new evidence to elucidate the many body quantum dynamics in dipolar spin glasses. PMID- 14754149 TI - Interference of magnetic and anisotropic tensor susceptibility reflections in resonant X-ray scattering of GdB4. AB - Resonant x-ray scattering experiments at the Gd L3 edge show interference between magnetic and anisotropic tensor susceptibility (ATS) reflections in GdB4. Energy profiles obtained from the magnetic and ATS resonances exhibited approximately 10 eV separation between the maximum resonance energies. The findings show that the Gd 5d band experienced hybridization giving rise to a significant split into isotropic lower energy band and distorted upper band states that account for the magnetic and ATS scattering, respectively. PMID- 14754150 TI - Interplay between disorder and quantum and thermal fluctuations in ferromagnetic alloys: the case of UCu2Si2-xGex. AB - We consider, theoretically and experimentally, the effects of structural disorder, quantum fluctuations, and thermal fluctuations in the magnetic and transport properties of certain ferromagnetic alloys. We study the particular case of UCu2Si2-xGex. The low temperature resistivity, rho(T,x), exhibits Fermi liquid behavior as a function of temperature T for all values of x, which can be interpreted as a result of the magnetic scattering of the conduction electrons from the localized U spins. The residual resistivity, rho(0,x), follows the behavior of a disordered binary alloy. The observed nonmonotonic dependence of the Curie temperature, T(c)(x), with x can be explained within a model of localized spins interacting with an electronic bath. Our results clearly show that the Curie temperature of certain alloys can be enhanced due to the interplay between quantum and thermal fluctuations with disorder. PMID- 14754151 TI - Vortex flux channeling in magnetic nanoparticle chains. AB - A detailed understanding of the formation of magnetic vortices in closely spaced ferromagnetic nanoparticles is important for the design of ultra-high-density magnetic devices. Here, we use electron holography and micromagnetic simulations to characterize three-dimensional magnetic vortices in chains of FeNi nanoparticles. We show that the diameters of the vortex cores depend sensitively on their orientation with respect to the chain axis and that vortex formation can be controlled by the presence of smaller particles in the chains. PMID- 14754152 TI - Magnetodielectric effects from spin fluctuations in isostructural ferromagnetic and antiferromagnetic systems. AB - We report on the effects of spin fluctuations, magnetic ordering, and external magnetic field on the dielectric constant of the ferromagnet SeCuO3, and the antiferromagnet TeCuO3. A model based on the coupling between uniform polarization and the q-dependent spin-spin correlation function is presented to explain the different behaviors for these isostructural compounds. The large magnetocapacitance near the transition temperature in the ferromagnet SeCuO3 suggests routes to enhancing the magnetodielectric response for practical applications. PMID- 14754153 TI - Current-induced two-level fluctuations in pseudo-spin-valve (Co/Cu/Co) nanostructures. AB - Two-level fluctuations of the magnetization state of pseudo-spin-valve pillars Co(10 nm)/Cu(10 nm)/Co(30 nm) embedded in electrodeposited nanowires ( approximately 40 nm in diameter, 6000 nm in length) are triggered by spin polarized currents of 10(7) A/cm(2) at room temperature. The statistical properties of the residence times in the parallel and antiparallel magnetization states reveal two effects with qualitatively different dependences on current intensity. The current appears to have the effect of a field determined as the bias field required to equalize these times. The bias field changes sign when the current polarity is reversed. At this field, the effect of a current density of 10(7) A/cm(2) is to lower the mean time for switching down to the microsecond range. This effect is independent of the sign of the current and is interpreted in terms of an effective temperature for the magnetization. PMID- 14754154 TI - Spectroscopy of few-electron collective excitations in charge-tunable artificial atoms. AB - We report the investigation of electronic excitations in InGaAs self-assembled quantum dots using resonant inelastic light scattering. The dots can be charged via a gate by N=1, em leader,6 electrons. We observe excitations, which are identified as transitions of electrons, predominantly from the s to the p shell (s-p transitions) of the quasiatoms. We find that the s-p transition energy decreases and the observed band broadens, when the p shell is filled with 1 to 4 electrons. By a theoretical model, which takes into account the full Coulomb interaction in the few-electron artificial atom, we can confirm the experimental results to be an effect of the Coulomb interaction in the quantum dot. PMID- 14754155 TI - Local chiral-symmetry breaking in globally centrosymmetric crystals. AB - A thorough tensor analysis of the Bragg-forbidden reflection (00.3)(h) in corundum systems having a global center of inversion, such as V2O3 and alpha Fe2O3, shows that anomalous x-ray resonant diffraction can access chiral properties related to the dipole-quadrupole (E1-E2) channel via an interference with the pure quadrupole-quadrupole (E2-E2) process. This is also confirmed by independent ab initio numerical simulations. In such a way, it becomes possible to detect chiral quantities in systems where dichroic absorption techniques are ineffective. PMID- 14754156 TI - Ferroelectricity in barium titanate quantum dots and wires. AB - Properties of BaTiO3 colloidal quantum dots and wires are simulated using a first principles-based approach. Large atomic off-center displacements (that are robust against capping matrix materials) are found to exist in very small (<5 nm) dots. We further determine the size dependences of electrical and electromechanical responses in the studied nanostructures, as well as provide microscopic understanding of these responses. PMID- 14754157 TI - Accuracy of circular polarization as a measure of spin polarization in quantum dot qubits. AB - A quantum dot spin light emitting diode provides a test of carrier spin injection into a qubit and a means for analyzing carrier spin injection and local spin polarization. Even with 100% spin-polarized carriers the emitted light may be only partially circularly polarized due to the geometry of the dot. We have calculated carrier polarization-dependent optical matrix elements for InAs/GaAs self-assembled quantum dots (SAQDs) for electron and hole spin injection into a range of quantum dot sizes and shapes, and for arbitrary emission directions. Calculations for typical SAQD geometries with emission along [110] show light that is only 5% circularly polarized for spin states that are 100% polarized along [110]. Measuring along the growth direction gives near unity conversion of spin to photon polarization and is the least sensitive to uncertainties in SAQD geometry. PMID- 14754158 TI - Eigenvector approximation leading to exponential speedup of quantum eigenvalue calculation. AB - We present an efficient method for preparing the initial state required by the eigenvalue approximation quantum algorithm of Abrams and Lloyd. Our method can be applied when solving continuous Hermitian eigenproblems, e.g., the Schrodinger equation, on a discrete grid. We start with a classically obtained eigenvector for a problem discretized on a coarse grid, and we efficiently construct, quantum mechanically, an approximation of the same eigenvector on a fine grid. We use this approximation as the initial state for the eigenvalue estimation algorithm, and show the relationship between its success probability and the size of the coarse grid. PMID- 14754159 TI - Teleportation in a noisy environment: a quantum trajectories approach. AB - We study the fidelity of quantum teleportation for the situation in which quantum logic gates are used to provide the long distance entanglement required in the protocol, and where the effect of a noisy environment is modeled by means of a generalized amplitude damping channel. Our results demonstrate the effectiveness of the quantum trajectories approach, which allows the simulation of open systems with a large number of qubits (up to 24). This shows that the method is suitable for modeling quantum information protocols in realistic environments. PMID- 14754161 TI - Effective interactions cannot replace solvent effects in a lattice model of proteins. AB - Protein folding and protein design are among the most challenging problems of the past ten years in biophysics and molecular biology. For a given protein, it is possible to extract, from existing protein databases, a set of specific (i.e., belonging to the investigated protein) effective amino-acid (AA) interactions able to stabilize the native state. On the other hand, attempts to find global effective AA interactions, which would be able to stabilize all proteins at once, failed. Using a simple lattice model where the solvent degrees of freedom are (semi)explicitly taken into account, we show that the absence of global effective AA interactions is due to the solvent and that on this lattice model the solvent effects cannot be reproduced by amino-acid effective interactions. PMID- 14754160 TI - Sheet excitability and nonlinear wave propagation. AB - In the Xenopus laevis oocyte, calcium ion channels are clustered in a thin shell. Motivated by this morphology, we study a general class of reaction-diffusion systems that include most of the well-known models that support wave propagation but restricting excitability to a "sheet" of codimension 1. We find waves that undergo propagation failure with increasing diffusion coefficient and a scaling regime in which the wave speed is independent of it. PMID- 14754162 TI - A motor that makes its own track: helicase unwinding of DNA. AB - We study the unwinding of DNA by helicase proteins as a representative system in which a motor protein interacts with a mobile obstacle. In our discrete model, the interaction between the helicase and the DNA fork is characterized by an interaction potential. For the case of a hard-wall potential, the helicase opens the DNA by rectifying thermal fluctuations which spontaneously open base pairs. A potential with nonzero range describes the destabilization of the double strand by the enzymatic action of the helicase. We derive solutions for the opening speed as a function of the potential shape and relate our results to experiments on helicase motion. PMID- 14754163 TI - Highly structured duets in the song of the South American Hornero. AB - The South American Hornero (Furnarius rufus) is a suboscine bird widely known for its mud-made, oven-looking nest. Beyond their architectural skills, the male and female Horneros sing in highly structured duets. The analysis of field recordings reported in this work reveals that as the male increases the note production rate the female responds by switching to different locking states: the ones predicted by the theory of nonlinear forced oscillators. This gives the duet a most appealing rhythm, and unveils the nonlinear nature of the underlying brain activity needed to generate the song. PMID- 14754164 TI - Experimental realization of a quantum spin pump. AB - We demonstrate the operation of a quantum spin pump based on cyclic radio frequency excitation of a GaAs quantum dot, including the ability to pump pure spin without pumping charge. The device takes advantage of bidirectional mesoscopic fluctuations of pumped current, made spin dependent by the application of an in-plane Zeeman field. Spin currents are measured by placing the pump in a focusing geometry with a spin-selective collector. PMID- 14754165 TI - Vibrational sum frequency scattering from a submicron suspension. AB - A novel application of vibrational sum frequency generation (VSFG) is developed to study the molecular properties of the surface of submicron particles in suspension. The Rayleigh-Gans-Debye scattering theory is extended to extract the local molecular response from the macroscopic nonlinearly scattered spectral intensity. These results demonstrate the use of VSFG to investigate quantitatively the surface molecular properties of submicron particles, dispersed in solution. It provides information on the order and density of alkane chains and allows us to determine the elements of the local second-order surface susceptibility. PMID- 14754166 TI - Comment on "Twist defect in chiral photonic structures". PMID- 14754168 TI - Projectile-shape dependence of impact craters in loose granular media. AB - We report on the penetration of cylindrical projectiles dropped from rest into a dry, noncohesive granular medium. The cylinder length, diameter, density, and tip shape are all explicitly varied. For deep penetrations, as compared to the cylinder diameter, the data collapse onto a single scaling law that varies as the 1/3 power of the total drop distance, the 1/2 power of cylinder length, and the 1/6 power of cylinder diameter. For shallow penetrations, the projectile shape plays a crucial role with sharper objects penetrating deeper. PMID- 14754169 TI - Structure and melting of two-species charged clusters in a parabolic trap. AB - We consider a system of charged particles interacting with an unscreened Coulomb repulsion in a two-dimensional parabolic confining trap. The static charge on a portion of the particles is twice as large as the charge on the remaining particles. The particles separate into a shell structure with those of greater charge situated farther from the center of the trap. As we vary the ratio of the number of particles of the two species, we find that for certain configurations, the symmetry of the arrangement of the inner cluster of singly charged particles matches the symmetry of the outer ring of doubly charged particles. These matching configurations have a higher melting temperature and a higher thermal threshold for intershell rotation between the species than the nonmatching configurations. PMID- 14754170 TI - Computationally efficient phase-field models with interface kinetics. AB - We present a phase-field model of solidification which allows efficient computations in the regime when interface kinetic effects dominate over capillary effects. The asymptotic analysis required to relate the parameters in the phase field with those of the original sharp-interface model is straightforward, and the resultant phase-field model can be used for a wide range of material parameters. PMID- 14754171 TI - Optical studies on free-standing films of an achiral smectic liquid crystal. AB - Employing null transmission ellipsometry and depolarized reflected light microscopy, we have studied two smectic phases, Sm-C1 and Sm-C2, of one achiral mesogen. Our results show that Sm-C1 and Sm-C2 are the synclinic Sm-C and anticlinic Sm-C(A) phases, respectively. We find no evidence to support recent claims that the Sm-C1 is chiral and ferroelectric [R. Stannarius et al., Phys. Rev. Lett. 90, 025502 (2003)]. PMID- 14754172 TI - Unusual behavior of the surface-induced tilted layers in free-standing films of a non-layer-shrinkage liquid crystal compound. AB - Null-transmission ellipsometry has been conducted to study the molecular arrangements in free-standing films of one chiral compound above the bulk smectic A-smectic-C* transition temperature. Upon cooling under a proper electric field, a nonplanar-anticlinic-synclinic or a nonplanar-synclinic transition has been observed. The nonplanar structure continuously evolves into the anticlinic or synclinic structures. Increasing electric field can induce a rare transition from a synclinic to an anticlinic structure. PMID- 14754173 TI - Structure of twist-grain-boundary-C phases. AB - We study properties of the Renn-Lubensky twist-grain-boundary-C (TGB(C)) phase, with layer normal rotating in a plane perpendicular to the pitch axis, and the Bordeaux TGB(C) phase, with the layer normal rotating on a cone parallel to the pitch axis near the upper critical twist k(c2) marking the transition to the cholesteric phase. We introduce a generalized model free energy for the smectic-C phase that allows either TGB(C) phase to be stable, and we calculate k(c2) and the order-parameter profile, which shows only modest spatial variation, for both phases. PMID- 14754174 TI - Stochastic resonance with spatiotemporal signal controlled by time delays. AB - Stochastic resonance in two coupled threshold elements with input periodic signals shifted in phase is studied. For fixed phase shift and coupling strength the signal-to-noise ratio at the output of each element can be maximized by introducing proper time delays in the coupling terms which cancel the effect of the phase shift. This shows that in systems of coupled elements driven by spatiotemporal periodic signals stochastic resonance can be controlled by delayed coupling. PMID- 14754175 TI - Dynamics of the depletion zone at a finite-sized imperfect trap in two dimensions: photobleaching experiments and simulations. AB - The kinetics of the growth of depletion zones around a static trap in an effective two-dimensional geometry were studied experimentally with photobleaching of fluorescein dye by a focused laser beam. The phototrap served as an imperfect trap with a finite size. The growth of the depletion zone was monitored by the theta distance, defined as the distance from the trap to the point where the concentration of the reactants reaches a given arbitrary fraction theta (0>1. The treatment is based on the gas-kinetic theory analysis of drag force in the specular and diffuse scattering limits obtained in a preceding paper [Z. Li and H. Wang, Phys. Rev. E., 68, 061206 (2003)]. Our analysis considers the influence of van der Waals interactions on the momentum transfer upon collision of a gas molecule with the particle and expresses this influence in terms of an effective, reduced collision integral. This influence is shown to be significant for nanosized particles. In the present paper, the reduced collision integral values are obtained for specular and diffuse scattering, using a Lennard-Jones-type potential energy function suitable for the interactions of a gas molecule with a particle. An empirical formula for the momentum accommodation function, used to determine the effective, reduced collision integral, is obtained from available experimental data. The resulting treatment is shown to be accurate for interpreting the mobility experiments for particles as small as approximately 1 nm in radius. The treatment is subsequently extended to the entire range of the Knudsen number, following a semiempirical, gas-kinetic theory analysis. We demonstrate that the proposed formula predicts very well Millikan's oil-droplet experiments [R. A. Millikan, Philos. Mag. 34, 1 (1917); Phys. Rev. 22, 1 (1923)]. The rigorous theoretical foundation of the proposed formula in the Kn>>1 limit makes the current theory far more general than the semiempirical Stokes-Cunningham formula in terms of the particle size and condition of the fluid and, therefore, more attractive than the Stokes Cunningham formula. PMID- 14754193 TI - Cage diffusion in liquid mercury. AB - We present inelastic neutron scattering measurements on liquid mercury at room temperature for wave numbers q in the range 0.3 infinity, the local theory predicts that the highest valency counterions density diverges as psi(nu) with some nu>1 or faster, then the electrostatic potential will saturate. Otherwise, if the counterion density diverges as psi or slower, or does not diverge as psi--> infinity, the electric potential will not saturate. Using this condition, we investigate the possibility of the saturation phenomenon within the framework of recent theories proposed in the literature to describe electrical double layer beyond the Poisson-Boltzmann description. PMID- 14754199 TI - Collective stochastic resonance in shear-induced melting of sliding bilayers. AB - The far-from-equilibrium dynamics of two crystalline two-dimensional monolayers driven past each other is studied using Brownian dynamics simulations. While at very high and low driving rates the layers slide past one another retaining their crystalline order, for intermediate range of drives the system alternates irregularly between the crystalline and fluidlike phases. A dynamical phase diagram in the space of interlayer coupling and drive is obtained. A qualitative understanding of this stochastic alternation between the liquidlike and crystalline phases is proposed in terms of a reduced model within which it can be understood as a stochastic resonance for the dynamics of collective order parameter variables. This remarkable example of stochastic resonance in a spatially extended system should be seen in experiments which we propose in the paper. PMID- 14754200 TI - Clustering and viscosity in a shear flow of a particulate suspension. AB - A shear flow of particulate suspension is analyzed for the qualitative effect of particle clustering on viscosity using a simple kinetic clustering model and direct numerical simulations. The clusters formed in a Couette flow can be divided into rotating chainlike clusters and layers of particles at the channel walls. The size distribution of the rotating clusters is scale invariant in the small-cluster regime and decreases rapidly above a characteristic length scale that diverges at a jamming transition. The behavior of the suspension can qualitatively be divided into three regimes. For particle Reynolds number Re(p) less than or approximately equal 0.1, viscosity is controlled by the characteristic cluster size deduced from the kinetic clustering model. For Re(p) approximately 1, clustering is maximal, but the simple kinetic model becomes inapplicable presumably due to onset of instabilities. In this transition regime viscosity begins to increase. For Re(p) greater than or approximately equal 10, inertial effects become important, clusters begin to breakup, and suspension displays shear thickening. This phenomenon may be attributed to enhanced contribution of solid phase in the total shear stress. PMID- 14754201 TI - Capillary condensation and interface structure of a model colloid-polymer mixture in a porous medium. AB - We consider the Asakura-Oosawa model of hard sphere colloids and ideal polymers in contact with a porous matrix modeled by immobilized configurations of hard spheres. For this ternary mixture a fundamental measure density functional theory is employed, where the matrix particles are quenched and the colloids and polymers are annealed, i.e., allowed to equilibrate. We study capillary condensation of the mixture in a small sample of matrix as well as demixing and the fluid-fluid interface inside a bulk matrix. Density profiles normal to the interface and surface tensions are calculated and compared to the case without matrix. Two kinds of matrices are considered: (i) colloid-sized matrix particles at low packing fractions and (ii) large matrix particles at high packing fractions. These two cases show fundamentally different behavior and should both be experimentally realizable. Furthermore, we argue that capillary condensation of a colloidal suspension could be experimentally accessible. We find that in case (ii), even at high packing fractions, the main effect of the matrix is to exclude volume and, to high accuracy, the results can be mapped onto those of the same system without matrix via a simple rescaling. PMID- 14754202 TI - Phase diagram of a quasi-two-dimensional colloid assembly. AB - We report the results of simulations of the phase diagrams of a quasi-two dimensional (Q2D) colloid assembly and of a two-dimensional (2D) colloid assembly which have the same colloid-colloid interaction. That interaction is the same as used in the study reported by Zangi and Rice [Phys. Rev. E 58, 7529 (1998)]. Among the goals of the work reported are elucidation of the influence of small amplitude out-of-plane motion on the phase diagram of a system and determination of the effect of that motion on the role of a hexatic phase in the melting process. Both of the systems we have studied undergo a first-order solid I-solid II and solid II-solid III isostructural transition induced by the attractive and repulsive components of the interaction, respectively. Introduction of the out-of plane motion shifts the low density portion of the phase boundaries involving the solid II phase. The liquid-solid I coexistence line is nearly the same for the two systems. The solid II-solid III transition is shifted to lower temperature and shifted to higher density in the quasi-two-dimensional system. We further use the simulations to calculate the elastic constants, which can be used to predict the location of the Kosterlitz-Thouless-Halperin-Nelson-Young (KTHNY) melting transition. For the Q2D system we find that the first-order melting transition preempts the KTHNY transition for the reduced temperatures T(*)=1.00, 0.60, and 0.50. For the 2D system, when T(*)=0.60, the KTHNY transition barely preempts the first-order melting transition and when T(*)=1.00 and 0.50 the ordinary first order transition preempts the KTHNY transition. PMID- 14754203 TI - Crystal structures of two-dimensional magnetic colloids in tilted external magnetic fields. AB - The stability of different crystal lattices of two-dimensional superparamagnetic suspensions that are confined to a planar liquid-gas interface and exposed to a tilted external magnetic field is studied theoretically by lattice sum minimizations. The magnetic field induces magnetic dipoles onto the colloidal particles along its direction, whose strength can be controlled by the amplitude of the external field. The mutual interaction between the colloids is governed by dipole-dipole forces and a short-ranged repulsion having its physical origin at the presence of the colloidal cores. If the direction of the magnetic field is perpendicular to the liquid-gas interface, there is a purely repulsive interaction leading to stable triangular crystals. By tilting the external field, the interaction becomes anisotropic and a mutual attraction appears upon a threshold tilt angle. We have calculated the full phase diagram at zero temperature varying the tilt angle, the colloidal density, and the strength of the magnetic field. Apart from the triangular lattice we find a variety of stable crystal lattices including rectangular, oblique, chainlike oblique, and rhombic structures. We also present the accurate derivation of the Hamiltonian of two polarizable particles of finite arbitrary geometries in external magnetic and electric fields. PMID- 14754204 TI - Dynamic density functional study of a driven colloidal particle in polymer solutions. AB - The dynamic density functional (DDF) theory and standard Brownian dynamics simulations (BDS) are used to study the drifting effects of a colloidal particle in a polymer solution, both for ideal and interacting polymers. The structure of the stationary density distributions and the total induced current are analyzed for different drifting rates. We find good agreement with the BDS, which gives support to the assumptions of the DDF theory. The qualitative aspect of the density distribution are discussed and compared to recent results for driven colloids in one-dimensional channels and to analytical expansions for the ideal solution limit. PMID- 14754205 TI - Average hydrodynamic correction for the Brownian dynamics calculation of flocculation rates in concentrated dispersions. AB - In order to account for the hydrodynamic interaction (HI) between suspended particles in an average way, Honig et al. [J. Colloid Interface Sci. 36, 97 (1971)] and more recently Heyes [Mol. Phys. 87, 287 (1996)] proposed different analytical forms for the diffusion constant. While the formalism of Honig et al. strictly applies to a binary collision, the one from Heyes accounts for the dependence of the diffusion constant on the local concentration of particles. However, the analytical expression of the latter approach is more complex and depends on the particular characteristics of each system. Here we report a combined methodology, which incorporates the formula of Honig et al. at very short distances and a simple local volume-fraction correction at longer separations. As will be shown, the flocculation behavior calculated from Brownian dynamics simulations employing the present technique, is found to be similar to that of Batchelor's tensor [J. Fluid. Mech. 74, 1 (1976); 119, 379 (1982)]. However, it corrects the anomalous coalescence found in concentrated systems as a result of the overestimation of many-body HI. PMID- 14754206 TI - Effects of asymmetric salt and a cylindrical macroion on charge inversion: electrophoresis by molecular dynamics simulations. AB - The charge inversion phenomenon is studied by molecular dynamics simulations, focusing on size and valence asymmetric salts, and a threshold of surface charge density for charge inversion. The charge inversion criteria by the electrophoretic mobility and the radial distribution functions of ions coincide except around the charge inversion threshold. The reversed electrophoretic mobility increases with the ratio of coion to counterion radii, a(-)/a(+), while it decreases with the ratio of coion to counterion valences, Z(-)/Z(+). The monovalent salt enhances charge inversion of a strongly charged macroion at small salt ionic strength, while it reduces reversed mobility otherwise. A cylindrical macroion is more persistent to monovalent salt than a spherical macroion of the same radius and surface charge density. PMID- 14754207 TI - Plastic flow in two-dimensional solids. AB - A time-dependent Ginzburg-Landau model of plastic deformation in two-dimensional solids is presented. The fundamental dynamic variables are the displacement field u and the lattice velocity v=delta(u)/delta(t). Damping is assumed to arise from the shear viscosity in the momentum equation. The elastic energy density is a periodic function of the shear and tetragonal strains, which enables the formation of slips at large strains. In this work we neglect defects such as vacancies, interstitials, or grain boundaries. The simplest slip consists of two edge dislocations with opposite Burgers vectors. The formation energy of a slip is minimized if its orientation is parallel or perpendicular to the flow in simple shear deformation and if it makes angles of +/-pi/4 with respect to the stretched direction in uniaxial stretching. High-density dislocations produced in plastic flow do not disappear even if the flow is stopped. Thus large applied strains give rise to structurally disordered states, which are metastable due to the Peierls potential. We divide the elastic energy into an elastic part due to affine deformation and a defect part. The latter represents degree of disorder and is nearly constant in plastic flow under cyclic straining. PMID- 14754208 TI - Simple analytic equations of state for hard-core single and double Yukawa fluids and mixtures based on second-order Barker-Henderson perturbation theory. AB - A simple analytic expression with high precision for the radial distribution function of hard spheres is proposed. The form of the expression has been carefully selected to combine the well-known Camahan-Starling equation of state in it and satisfy the limit condition at low density, its simplicity and precision is superior to the well-known Percus-Yevick expression. The coefficients contained in the expression have been determined by fitting the Monte Carlo data for the first coordination shell, and by fitting both the Monte Carlo data and the numerical results of the Percus-Yevick expression for the second coordination shell. The expression has been applied to develop simple analytic equations of state for the hard-core single, double Yukawa fluids, and the hard-core Yukawa mixtures. The comparisons show that the agreement of our model with the computer simulation data is slightly better than the mean spherical approximation and other analytic models. PMID- 14754209 TI - Local mean-field study of capillary condensation in silica aerogels. AB - We apply local mean-field (i.e., density functional) theory to a lattice model of a fluid in contact with a dilute, disordered gel network. The gel structure is described by a diffusion-limited cluster aggregation model. We focus on the influence of porosity on both the hysteretic and the equilibrium behavior of the fluid as one varies the chemical potential at low temperature. We show that the shape of the hysteresis loop changes from smooth to rectangular as the porosity increases and that this change is associated with disorder-induced out-of equilibrium phase transitions that differ in adsorption and in desorption. Our results provide insight in the behavior of 4He in silica aerogels. PMID- 14754210 TI - Statistical analysis of two-dimensional cluster structures composed of ferromagnetic particles based on a flexible chain model. AB - We investigate two-dimensional cluster structures composed of ferromagnetic colloidal particles, based on a flexible chain model, by the configurational-bias Monte Carlo method. We clarify the dependence of the probabilities of the creation of different types of clusters on the dipole-dipole interactive energy and the cluster size. PMID- 14754211 TI - Structure and phase behavior of a two-dimensional system with core-softened and long-range repulsive interactions. AB - The structure and phase behavior of a two-dimensional system with purely repulsive core-softened and long-range interactions are studied using Monte Carlo computer simulations. The pair interactions are of the form, u(r)=4epsilon[(sigma/r)(12)-(sigma/r)(6)]+epsilon(')(sigma/r)(3), with the energy parameter, epsilon(')=(8sqrt[6]/9)epsilon chosen to give a stationary point of inflection in the pair potential at r=6(1/6)sigma. This potential approximates the effective interparticle interactions for a two-dimensional dipolar system in a strong field aligned perpendicular to the plane. The low-temperature portion of the phase diagram is sketched out, and the static properties of the various phases are analyzed in some detail. At low temperatures a variety of interesting states are in evidence, including: fluids with chainlike, striped, and 6-10 sided polygon structural motifs; low-density and high-density triangular crystalline phases; and defective Kagome lattices. It is shown that clustering is driven by the presence of the repulsive shoulder in the pair potential. Other features, such as the presence of a disordered phase with a network structure, are due to the long-range (1/r(3)) repulsive tail in the potential. The relevance of the simulation results to experimental work, including materials synthesis, is briefly discussed. PMID- 14754212 TI - Dynamics of shear-transformation zones in amorphous plasticity: energetic constraints in a minimal theory. AB - We use energetic considerations to deduce the form of a previously uncertain coupling term in the shear-transformation-zone (STZ) theory of plastic deformation in amorphous solids. As in the earlier versions of the STZ theory, the onset of steady deformation at a yield stress appears here as an exchange of dynamic stability between jammed and plastically deforming states. We show how an especially simple "quasilinear" version of this theory accounts qualitatively for many features of plasticity such as yielding, strain softening, and strain recovery. We also show that this minimal version of the theory fails to describe certain other phenomena, and argue that these limitations indicate needs for additional internal degrees of freedom beyond those included here. PMID- 14754213 TI - Freezing transition and correlated motion in a quasi-two-dimensional colloid suspension. AB - Recent experiments have demonstrated that the deviation of the single-particle displacement distribution from Gaussian form in a dense quasi-two-dimensional colloid suspension is a result of heterogenous dynamics that involves cooperative motions of neighboring colloid particles [J. Chem. Phys. 47, 9142 (2001)]. In this paper, we report the results of molecular dynamics (MD) simulations of a quasi-two-dimensional assembly of nearly hard-sphere colloid particles. The colloid-colloid interaction we use is short ranged and everywhere repulsive; it is related to the Marcus-Rice (MR) and modified MR interactions used in a previous study [Phys. Rev. E 58, 7529 (1998)]. As is the case for those systems, the one we study supports liquid, hexatic, and solid phases. Our calculations show that the deviation of the single-particle displacement distribution from Gaussian form is present in the liquid phase, and that a sharp increase in its magnitude occurs at the liquidus density and extends into the crystalline phase. For densities greater than the liquidus density we find three dynamical relaxation processes that include, at intermediate times, a slowing down in the rate of growth of the diffusive displacement of a particle due to the cage effect. As the density increases toward the solidus density, the dependence of the mean squared displacement on time, at intermediate times, changes from sublinear to zero. The onset of the long-time relaxation mode corresponds to the time at which the deviation of the particle displacement distribution from Gaussian form is a maximum. At this time, which increases exponentially with the density, the self-part of the van Hove function exhibits multiple maxima with respect to r while the distinct part of the van Hove function is a maximum at the origin, thereby signaling jump dynamics. At long times the particle mean square displacement has diffusive character at all densities including solid phase densities. A remarkable feature of our findings is the continuity of character of the particle displacement from the liquid phase through the hexatic phase and into the solid phase. Cooperative jumps that lead to diffusive process in crystals can be explained by a mechanism that involves many such correlated hops in random locations and random directions (but along the crystallographic axes) thereby generating effective random walk behavior. We argue that the collective motion we have found is generated by superpositions of instantaneous normal mode vibrations along diffusive paths. The diffusive paths are along the directions with strong bond orientation correlation, and start to grow in amplitude rapidly on entry into the hexatic phase. PMID- 14754214 TI - Fractal patterns, cluster dynamics, and elastic properties of magnetorheological suspensions. AB - We study pattern formation and the aggregation processes in magnetorheological suspensions in the presence of a static magnetic field, and some of their associated physical properties. In particular, we analyze the elastic modes as a function of the intensity of the applied field and for several particle concentrations. We observe that the clusters formed in these systems have multifractal characteristics, which are the result of three well defined stages of the aggregation process. In these stages three generations of clusters are produced sequentially. The structure of the suspension can be well characterized by its mass fractal dimensions and the mass radial distribution. The size distribution of the second-generation clusters written in terms of their mass fractal dimension allows us to calculate the sound speed of the longitudinal modes in the large wavelength regime. This multifractal analysis applied to several kinds of aggregates reveals that the occurrence of at least three stages of aggregation is a common feature to several physical aggregation processes. PMID- 14754215 TI - Phase diagrams of classical spin fluids: the influence of an external magnetic field on the liquid-gas transition. AB - The influence of an external magnetic field on the liquid-gas phase transition in Ising, XY, and Heisenberg spin fluid models is studied using a modified mean field theory and Gibbs ensemble Monte Carlo simulations. It is demonstrated that the theory is able to reproduce quantitatively all characteristic features of the field dependence of the critical temperature T(c)(H) for all the three models. These features include a monotonic decrease of T(c) with rising H in the case of the Ising fluid as well as a more complicated nonmonotonic behavior for the XY and Heisenberg models. The nonmonotonicity consists in a decrease of T(c) with increasing H at weak external fields, an increase of T(c) with rising H in the strong field regime, and the existence of a minimum in T(c)(H) at intermediate values of H. Analytical expressions for T(c)(H) in the large field limit are presented as well. The paramagnetic-ferromagnetic phase transition is also considered in simulations and described within the mean field theory. PMID- 14754216 TI - Lattice study of a Janus interface. AB - A lattice gas simulation of water between a hydrophobic plate and a hydrophilic plate (a Janus interface) shows large fluctuations in the number of liquid cells in contact with the hydrophobic plate, and a power spectrum similar to the experimental results that Zhang, Zhu, and Granick found [X. Y. Zhang, Y. X. Zhu, and S. Granick, Science 295, 663 (2002)] when measuring viscous response in a Janus system. Study of the spatial Fourier modes of the liquid-vapor interface suggests that interface fluctuations with length scales between approximately 1.5 and 20 nm cause the effects observed in the simulation. PMID- 14754217 TI - Anisotropy of alkyl chains of azobenzene molecules at the air/water interface observed by sum-frequency vibrational spectroscopy. AB - Surface-specific sum-frequency vibrational spectroscopy has been used to study the structure of alkyl chains of azobenzene molecules at the air/water interface. The results show that the alkyl chains are well aligned before UV irradiation and protruding out of the surface with a certain distribution. Although the alkyl chains become less ordered by UV irradiation following dynamical motion due to cis-trans isomerization of the azobenzene core, the alkyl chains show anisotropy in the direction perpendicular to that of the azobenzene core by linearly polarized UV irradiation. PMID- 14754218 TI - Spreading dynamics of polymer nanodroplets. AB - The spreading of polymer droplets is studied using molecular dynamics simulations. To study the dynamics of both the precursor foot and the bulk droplet, large hemispherical drops of 200 000 monomers are simulated using a bead spring model for polymers of chain length 10, 20, and 40 monomers per chain. We compare spreading on flat and atomistic surfaces, chain length effects, and different applications of the Langevin and dissipative particle dynamics thermostats. We find diffusive behavior for the precursor foot and good agreement with the molecular kinetic model of droplet spreading using both flat and atomistic surfaces. Despite the large system size and long simulation time relative to previous simulations, we find that even larger systems are required to observe hydrodynamic behavior in the hemispherical spreading droplet. PMID- 14754219 TI - Dislocation loop dynamics in freestanding smectic films: the role of the disjoining pressure and of the finite permeability of the meniscus. AB - When a dislocation loop nucleates in a freestanding film, it collapses or grows depending on whether its radius r is smaller or larger than a critical radius r(c). In this paper, we analyze the growth dynamics of a dislocation loop in the limit of r>>r(c). Experiments with pure octylcyanobiphenyl show that the dislocation velocity is constant in thick films (more than 100 layers) regardless of their thicknesses, and only depends on the pressure in the meniscus. At intermediate thickness (between 100 and 15 layers), the velocity is no longer constant and tends to decrease in time on account of the finite permeability of the meniscus. In very thin films (less than 15 layers), the dislocations move faster than in thick films, although their velocities continue to decrease in time. The thinner the film, the larger the global acceleration is. This effect is linked to a supplementary force acting on the dislocations caused by the attraction between the free surfaces (where the smectic order parameter is enhanced). The progressive deceleration is due to the finite permeability of the meniscus. PMID- 14754220 TI - Anchoring properties of photoaligned azo-dye materials. AB - The photoinduced alignment of a liquid crystal (LC) on films of azo dyes was studied for the liquid crystal display applications. In order to improve the alignment stability of the photoaligned dye (SD-1), the azo-dye derivative with polymerizable terminal groups was synthesized (SDA-1). The films of SDA-1 exhibit good photoalignment properties and ultraviolet stability after thermal polymerization. We investigated the anchoring coefficient for polar and azimuthal energy for both azo-dyes SD-1 and SDA-1 using a differential method developed by us. The polar and azimuthal anchoring strengths of azo-dye aligning materials are comparable with those of usual polyimide aligning materials. The electro-optic response of a 90 degrees twisted LC cell remains almost the same after heating up to 200 degrees C, i.e., the anchoring energy remains considerably high. PMID- 14754221 TI - Free planar isotropic-nematic interfaces in binary hard-rod fluids. AB - Within the Onsager theory we study free planar isotropic-nematic interfaces in binary mixtures of hard rods. For sufficiently different particle shapes the bulk phase diagrams of these mixtures exhibit a triple point, where an isotropic (I) phase coexists with two nematic phases (N1 and N2) of different composition. For all explored mixtures we find that upon approach of the triple point the I-N2 interface shows complete wetting by an intervening N1 film. We compute the surface tensions of isotropic-nematic interfaces, and find a remarkable increase with fractionation, similar to the effect in polydisperse hard-rod fluids. PMID- 14754222 TI - Shear-induced textural transitions in flow-aligning liquid crystal polymers. AB - The equations of nematodynamics are formulated, solved, and used to model textural transformations in sheared thermotropic flow-aligning nematic polymers. The solutions are classified and characterized using analytical, scaling, and numerical methods. It is found that as the shear rate increases, the pathway between an oriented nonplanar state and an oriented planar state is through texture formation and coarsening. The two shear-rate dependent dimensionless numbers that control the texture formation and coarsening process are Ericksen Er and Deborah De numbers. The emergence of texture is independent of the Deborah number, and occurs at Er=10(4). As the shear rate increases and Er>10(4) the first texture that arises is a defect lattice. Further increases of the shear rate bring De close to 1, ignite the coarsening processes, and replace the defect lattice with a defect gas. The smallest texture length scale l(t) occurs at the defect lattice-defect gas transition. In the defect lattice regime the texture length scale decreases with increasing shear rate as l(t) proportional to (gamma a)(-1/2), while in the defect gas regime it increases as l(t) proportional to (gamma-b sqrt[(gamma-a)]-c)(-1). Finally when De>2, an oriented monodomain state emerges, and the texture vanishes since coarsening overpowers defect nucleation. It is found that the texture transition cascade unoriented monodomain=>defect lattice=>defect gas=>oriented monodomain is remarkably consistent with the experimentally observed textural transitions of sheared lyotropic nematic polymers. PMID- 14754223 TI - Effect of spontaneous polarization and polar surface anchoring on the director and layer structure in surface-stabilized ferroelectric liquid crystal cells. AB - We use the Landau-de Gennes model to study theoretically the effect of the magnitude of the spontaneous polarization (P(S)), the ratio (r) between the equilibrium layer tilt and the smectic cone angle, the thickness of the insulating alignment layers and the strength of the polar and nonpolar surface anchoring on the director and layer structure in surface-stabilized ferroelectric liquid crystal cells with the chevron structure of smectic layers. The system shows a surprising number of stable structures, accompanied by one or two metastable ones. At P(S) greater than the critical value only quasimonostable structures, which can exhibit the thresholdless (V-shaped) switching, exist at all r and both at weak and strong polar surface anchoring. At lower P(S) bistable and monostable structures can coexist. Bistable structures can be expected at high r, low P(S) and if polar surface anchoring is weaker than the nonpolar one. Lowering the ratio r and/or increasing the strength of polar anchoring promotes the stability of monostable structures. Thicker insulating alignment layers also drive the system into the monostable state. Polar surface anchoring induces a large surface electroclinic effect. As a result the nematic deformations close to the surfaces are very strong and the stress is relieved by bending of the smectic layers. This leads to the formation of a double chevron structure which is stable at very large polar anchoring and due to the surface electroclinic effect it is metastable also at lower values of polar anchoring. PMID- 14754224 TI - Identification of nonmonotonic behaviors and stick-slip transition in liquid crystal polymers. AB - The recent identification of shear-induced phases in the isotropic melts of liquid crystal polymers shows that these materials are expected to display original nonlinear behaviors. We have investigated the flow behavior of a nematic sidechain polymer above its isotropic-nematic transition temperature. Nonlinear rheology and birefringence measurements indicate the appearance, above a critical shear rate, of the shear-induced isotropic-nematic phase transition. The rheological behavior of this induced phase is characterized by undamped time periodic shear stress oscillations. These sustained oscillations are interpreted in terms of a stick-slip mechanism alternating high-friction static state and low friction kinetic state. PMID- 14754225 TI - Waves at the nematic-isotropic interface: the role of surface tension anisotropy, curvature elasticity, and backflow effects. AB - Recently, a theoretical description of waves at the nematic-isotropic interface has been proposed using a generalized dynamical Landau-Ginzburg-de Gennes theory [V. Popa-Nita and T. J. Sluckin, Phys. Rev. E 66, 041703 (2002)]. This calculation assumed an isotropic surface tension, i.e., independent of the director orientation at the interface and neglected all coupling between the director and the hydrodynamic flow. As a consequence, the director was assumed to keep a fixed orientation and do not couple with the oscillations of the interface. These assumptions are rather crude in real nematics where surface tension anisotropy may be as large as 20% and where hydrodynamic coupling with the director is known to be important. In this paper we propose to take into account these two effects: as a result, interface oscillations couple with the director field via hydrodynamic flows and backflow effects. We analyze how these phenomena change the dispersion relation. Finally, we review experiments on the nematic-isotropic interface and discuss how to measure experimentally the dispersion relation. PMID- 14754226 TI - Pulling reptating polymers by one end: magnetophoresis in the Rubinstein-Duke model. AB - We consider the magnetophoresis problem within the Rubinstein-Duke model, i.e., a reptating polymer pulled by a constant field applied to a single end of a chain. Extensive density matrix renormalization calculations are presented of the drift velocity and the profile of the chain for various strengths of the driving field and chain lengths. We show that the velocities and the average densities of the stored length are well described by simple interpolating crossover formulas, derived under the assumption that the difference between the drift and curvilinear velocities vanishes for sufficiently long chains. The profiles, which describe the average shape of the reptating chain, also show such interesting features as some nonmonotonic behavior of the link densities for sufficiently strong pulling fields. We develop a description in which a distinction is made between links entering at the pulled head and at the unpulled tail. At weak fields the separation between the head zone and the tail zone meanders through the whole chain, while the probability of finding it close to the edges drops off. At strong fields the tail zone is confined to a small region close to the unpulled edge of the polymer. PMID- 14754227 TI - Theory of polyelectrolytes in solvents. AB - Using a continuum description, we account for fluctuations in the ionic solvent surrounding a Gaussian, charged chain and derive an effective short-ranged potential between the charges on the chain. This potential is repulsive at short separations and attractive at longer distances. The chemical potential can be derived from this potential. When the chemical potential is positive, it leads to a meltlike state. For a vanishingly low concentration of segments, this state exhibits scaling behavior for long chains. The Flory exponent characterizing the radius of gyration for long chains is calculated to be approximately 0.63, close to the classical value obtained for second order phase transitions. For short chains, the radius of gyration varies linearly with N, the chain length, and is sensitive to the parameters in the interaction potential. The linear dependence on the chain length N indicates a stiff behavior. The chemical potential associated with this interaction changes sign, when the screening length in the ionic solvent exceeds a critical value. This leads to condensation when the chemical potential is negative. In this state, it is shown using the mean-field approximation that spherical and toroidal condensed shapes can be obtained. The thickness of the toroidal polyelectrolyte is studied as a function of the parameters of the model, such as the ionic screening length. The predictions of this theory should be amenable to experimental verification. PMID- 14754228 TI - Modeling charge transfer in the photosynthetic reaction center. AB - In this work, we present a model to elucidate the unidirectionality of the primary charge-separation process in the bacterial reaction centers. We have used a model of three sites/molecules with electron transfer beginning at site 1 with an option to proceed to site 2 or site 3. We used a stochastic model with arbitrary correlation functions. We get the quantum yields of electron escape via the sites 2,3 in two limiting cases that correspond to a spectral density of underdamped and overdamped Brownian oscillator. In the fast modulation limit of an overdamped regime we get the effect, which was named "fear of death," in which for strong enough sink parameters the electron has a tendency to avoid the place with greater sink. The presented model was used to provide a plausible explanation of the temperature dependence of the quantum yields of the Rhodobacter sphaeroides photosynthetic reaction center in the high-temperature regime. PMID- 14754229 TI - Analytical solution of a neutral model of biodiversity. AB - The unified neutral model of biodiversity proposed by S. Hubbell is solved analytically: The distributions of species abundance in the metacommunity and in a local community are calculated exactly as a function of speciation and migration rates and of the size of the community. In the limit of large population sizes the densities of species of given relative abundance are found to be given by universal functions depending only on two parameters. PMID- 14754230 TI - Adsorption of monovalent and multivalent cations and anions on DNA molecules. AB - Adsorption of monovalent and multivalent cations and anions on a deoxyribose nucleic acid (DNA) molecule from a salt solution is investigated by computer simulation. The ions are modeled as charged hard spheres, the DNA molecule as a point charge pattern following the double-helical phosphate strands. The geometrical shape of the DNA molecules is modeled on different levels ranging from a simple cylindrical shape to structured models which include the major and minor grooves between the phosphate strands. The densities of the ions adsorbed on the phosphate strands in the major and in the minor grooves are calculated. First, we find that the adsorption pattern on the DNA surface depends strongly on its geometrical shape: counterions adsorb preferentially along the phosphate strands for a cylindrical model shape, but in the minor groove for a geometrically structured model. Second, we find that an addition of monovalent salt ions results in an increase of the charge density in the minor groove while the total charge density of ions adsorbed in the major groove stays unchanged. The adsorbed ion densities are highly structured along the minor groove while they are almost smeared along the major groove. Furthermore, for a fixed amount of added salt, the major-groove cationic charge is independent of the counterion valency. For increasing salt concentration the major groove is neutralized while the total charge adsorbed in the minor groove is constant. DNA overcharging is detected for multivalent salts. Simulations for larger ion radii, which mimic the effect of ion hydration, indicate an increased adsorbtion of cations in the major groove. PMID- 14754231 TI - Hierarchical population model with a carrying capacity distribution for bacterial biofilms. AB - In order to describe biological colonies with a conspicuous hierarchical structure, a time- and space-discrete model for the growth of a rapidly saturating local biological population N(x,t) is derived from a hierarchical random deposition process previously studied in statistical physics. Two biologically relevant parameters, the probabilities of birth, B, and of death, D, determine the carrying capacity K. Due to the randomness the population depends strongly on position x and there is a distribution of carrying capacities, Pi(K). This distribution has self-similar character owing to the exponential slowing down of the growth, assumed in this hierarchical model. The most probable carrying capacity and its probability are studied as a function of B and D. The effective growth rate decreases with time, roughly as in a Verhulst process. The model is possibly applicable, for example, to bacteria forming a "towering pillar" biofilm, a structure poorly described by standard Eden or diffusion limited-aggregation models. The bacteria divide on randomly distributed nutrient rich regions and are exposed to a random local bactericidal agent (antibiotic spray). A gradual overall temperature or chemical change away from optimal growth conditions reduces bacterial reproduction, while biofilm development degrades antimicrobial susceptibility, causing stagnation into a stationary state. PMID- 14754232 TI - Budding of crystalline domains in fluid membranes. AB - Crystalline domains embedded in fluid membrane vesicles are studied by Monte Carlo simulations of dynamically triangulated surfaces and by scaling arguments. A budding transition from a caplike state to a budded shape is observed for increasing spontaneous curvature C0 of the crystalline domain as well as increasing line tension lambda. The location of the budding transition is determined as a function of C0, lambda, and the radius R(A) of the crystalline domain. In contrast to previous theoretical predictions, it is found that budding occurs at a value of the spontaneous curvature C0, that is always a decreasing function of the domain size R(A). Several characteristic scaling regimes are predicted. The distribution of five- and sevenfold disclinations as the budding transition is approached is determined, and the dynamics of the generation of defects is studied. PMID- 14754233 TI - Motion of a rotatory molecular motor and the chemical reaction rate. AB - We examine the dependence of the physical quantities of the rotatory molecular motor, such as the rotation velocity and the proton translocation rate, on the chemical reaction rate using the model based only on diffusion process. A peculiar behavior of proton translocation is found and the energy transduction efficiency of the motor protein is enhanced by this behavior. We give a natural explanation that this behavior is universal when certain inequalities between chemical reaction rates hold. That may give a clue to examine whether the motion of the molecular motor is dominated by diffusion process or not. PMID- 14754234 TI - Distinct regimes of elastic response and deformation modes of cross-linked cytoskeletal and semiflexible polymer networks. AB - Semiflexible polymers such as filamentous actin (F-actin) play a vital role in the mechanical behavior of cells, yet the basic properties of cross-linked F actin networks remain poorly understood. To address this issue, we have performed numerical studies of the linear response of homogeneous and isotropic two dimensional networks subject to an applied strain at zero temperature. The elastic moduli are found to vanish for network densities at a rigidity percolation threshold. For higher densities, two regimes are observed: one in which the deformation is predominately affine and the filaments stretch and compress; and a second in which bending modes dominate. We identify a dimensionless scalar quantity, being a combination of the material length scales, that specifies to which regime a given network belongs. A scaling argument is presented that approximately agrees with this crossover variable. By a direct geometric measure, we also confirm that the degree of affinity under strain correlates with the distinct elastic regimes. We discuss the implications of our findings and suggest possible directions for future investigations. PMID- 14754235 TI - Modeling the concentration-dependent permeation modes of the KcsA potassium ion channel. AB - The potassium channel from Streptomyces lividans (KcsA) is an integral membrane protein with sequence similarity to all known potassium channels, particularly in the selectivity filter region. A recently proposed model for ion channels containing either n or (n-1) single-file ions in their selectivity filters [P. H. Nelson, J. Chem. Phys. 177, 11396 (2002)] is applied to published KcsA channel K+ permeation data that exhibit a high-affinity process at low concentrations and a low-affinity process at high concentrations [M. LeMasurier et al., J. Gen. Physiol. 118, 303 (2001)]. The kinetic model is shown to provide a reasonable first-order explanation for both the high- and low-concentration permeation modes observed experimentally. The low-concentration mode ([K+]<200 mM) has a 200-mV dissociation constant of 56 mM and a conductance of 88 pS. The high-concentration mode ([K+]>200 mM) has a 200-mV dissociation constant of 1100 mM and a conductance of 500 pS. Based on the permeation model, and x-ray analysis [J. H. Morais-Cabral et al., Nature (London) 414, 37 (2001)], it is suggested that the experimentally observed K+ permeation modes correspond to an n=3 mechanism at high concentrations and an n=2 mechanism at low concentrations. The ratio of the electrical dissociation distances for the high- and low-concentration modes is 3:2, also consistent with the proposed n=3 and n=2 modes. Model predictions for K+ channels that exhibit asymmetric current-voltage (I-V) curves are presented, and further validation of the kinetic model via molecular simulation and experiment is discussed. The qualitatively distinct I-V characteristics exhibited experimentally by Tl+, NH+4, and Rb+ ions at 100 mM concentration can also be explained using the model, but more extensive experimental tests are required for quantitative validation of the model predictions. PMID- 14754236 TI - Thermal denaturation of a helicoidal DNA model. AB - We study the static and dynamical properties of DNA in the vicinity of its melting transition, i.e., the separation of the two strands upon heating. The investigation is based on a simple mechanical model which includes the helicoidal geometry of the molecule and allows an exact numerical evaluation of its thermodynamical properties. Dynamical simulations of long-enough molecular segments allow the study of the structure factors and of the properties of the denaturated regions. Simulations of finite chains display the hallmarks of a first order transition for sufficiently long-ranged stacking forces although a study of the model's "universality class" strongly suggests the presence of an "underlying" continuous transition. PMID- 14754237 TI - Universality and Shannon entropy of codon usage. AB - The distribution functions of codon usage probabilities, computed over all the available GenBank data for 40 eukaryotic biological species and five chloroplasts, are best fitted by the sum of a constant, an exponential, and a linear function in the rank of usage. For mitochondria the analysis is not conclusive. These functions are characterized by parameters that strongly depend on the total guanine and cytosine (GC) content of the coding regions of biological species. It is predicted that the codon usage is the same in all exonic genes with the same GC content. The Shannon entropy for codons, also strongly dependent on the exonic GC content, is computed. PMID- 14754238 TI - Reparametrizing the loop entropy weights: effect on DNA melting curves. AB - Recent advances in the understanding of the melting behavior of double-stranded DNA with statistical mechanics methods lead to improved estimates of the weight factors for the dissociation events of the chains, in particular for interior loop melting. So far, in the modeling of DNA melting, the entropy of denaturated loops has been estimated from the number of configurations of a closed self avoiding walk. It is well understood now that a loop embedded in a chain is characterized by a loop closure exponent c which is higher than that of an isolated loop. Here we report an analysis of DNA melting curves for sequences of a broad range of lengths (from 10 to 10(6) base pairs) calculated with a program based on the algorithms underlying MELTSIM. Using the embedded loop exponent we find that the cooperativity parameter is one order of magnitude bigger than current estimates. We argue that in the melting region the double helix persistence length is greatly reduced compared to its room temperature value, so that the use of the embedded loop closure exponent for real DNA sequences is justified. PMID- 14754239 TI - Clustering in neutral ecology. AB - The "neutral ecology" model assumes that all organisms in the same trophic level have the same death, duplication, migration and mutation rates and are subjected to a zero sum rule. We show by exact analytical methods that under the assumption of this model, organisms tend to aggregate and form clusters. At dimensions less than or equal to 2, cluster size grows in average and one specie will dominate the whole ecosystem if enough time is allocated. At dimension d=3 (or higher), an equilibrium is reached and cluster sizes are distributed as a power law. PMID- 14754240 TI - Nonlinear analysis of correlations in Alu repeat sequences in DNA. AB - We report on a nonlinear analysis of deterministic structures in Alu repeats, one of the richest repetitive DNA sequences in the human genome. Alu repeats contain the recognition sites for the restriction endonuclease AluI, which is what gives them their name. Using the nonlinear prediction method developed in chaos theory, we find that all Alu repeats have novel deterministic structures and show strong nonlinear correlations that are absent from exon and intron sequences. Furthermore, the deterministic structures of Alus of younger subfamilies show panlike shapes. As young Alus can be seen as mutation free copies from the "master genes," it may be suggested that the deterministic structures of the older subfamilies are results of an evolution from a "panlike" structure to a more diffuse correlation pattern due to mutation. PMID- 14754241 TI - Robustness of retrieval properties against imbalance between long-term potentiation and depression of spike-timing-dependent plasticity. AB - Spike-timing-dependent plasticity (STDP) has recently been shown in some physiological studies. STDP depends on the precise temporal relationship of presynaptic and postsynaptic spikes. Many authors have indicated that a precise balance between long-term potentiation (LTP) and long-term depression (LTD) of STDP is significant for a stable learning. However, a situation in which the balance is maintained precisely is inconceivable in the brain. Using a method of the statistical neurodynamics, we show robust retrieval properties of spatiotemporal patterns in an associative memory model against the imbalance between LTP and LTD. When the fluctuation of LTD is assumed to obey a Gaussian distribution with mean 0 and variance delta(2), the storage capacity takes a finite value even at large delta. This means that the balance between LTP and LTD of STDP need not be maintained precisely, but must be maintained on average. Furthermore, we found that the basin of attraction becomes smaller as delta increases while an initial critical overlap remains unchanged. PMID- 14754242 TI - Lipid membranes with free edges. AB - Lipid membrane with freely exposed edge is regarded as smooth surface with curved boundary. Exterior differential forms are introduced to describe the surface and the boundary curve. The total free energy is defined as the sum of Helfrich's free energy and the surface and line tension energy. The equilibrium equation and boundary conditions of the membrane are derived by taking the variation of the total free energy. These equations can also be applied to the membrane with several freely exposed edges. Analytical and numerical solutions to these equations are obtained under the axisymmetric condition. The numerical results can be used to explain recent experimental results obtained by Saitoh et al. [Proc. Natl. Acad. Sci. 95, 1026 (1998)]. PMID- 14754243 TI - Two-electron transfer reactions in proteins: bridge-mediated and proton-assisted processes. AB - Nonadiabatic two-electron transfer (TET) reactions through donor-bridge-acceptor (DBA) systems is investigated within the approximation of fast vibrational relaxation. For TET reactions in which the population of bridging states remains small (less than 10(-2)) it is demonstrated that a multiexponential transition process reduces to three-state kinetics. The transfer starts at the state with two excess electrons at the D center (D(2-)BA), goes through the intermediate (transient) state with one electron at the D center and one at the A center (D-BA ), and ends up with the two electrons at the A center (DBA2-). Furthermore, if the population of the intermediate state becomes also small the two-exponential kinetics can be transformed with high accuracy to single-exponential D-A TET kinetics. The related overall transfer rate contains contributions from stepwise and from concerted TET. The latter process is determined by a specific two electron superexchange coupling incorporating the bridging states (D-B-A and DB-A ) as well as the intermediate state (D-BA-). As an example, the reduction of micothione reductase by nicotinamide adenine dinucleotide phosphate is analyzed. Existing experimental data can be explained if one assumes that the proton assisted reduction of the enzyme is realized by the concerted TET mechanism. PMID- 14754244 TI - Phase switching in a system of two noisy Hodgkin-Huxley neurons coupled by a diffusive interaction. AB - The focus of this paper is on the synchronous activity of a system of two intrinsically noisy Hodgkin-Huxley neurons coupled by a diffusive interaction. It is shown that conductance noise allows the relative phase of the neurons to display several different dynamical regimes ranging from phase and antiphase locking to random switching between two or more states. A synchronization diagram displaying the structure of the distribution function of the cyclic relative phase of the two neurons is presented. The addition of sinusoidal forcing terms to the equations governing the membrane voltage of both neurons gives rise to the statistical locking of those random switchings to the phase of the external signal. PMID- 14754245 TI - Density functional theory and demixing of binary hard-rod-polymer mixtures. AB - A density functional theory for a mixture of hard rods and polymers modeled as chains built of hard tangent spheres is proposed by combining the functional due to Yu and Wu for the polymer mixtures [J. Chem. Phys. 117, 2368 (2002)] with Schmidt's functional [Phys. Rev. E 63, 50 201 (2001)] for rod-sphere mixtures. As a simple application of the functional, the demixing transition into polymer-rich and rod-rich phases is examined. When the chain length increases, the phase boundary broadens and the critical packing fraction decreases. The shift of the critical point of a demixing transition is most noticeable for short chains. PMID- 14754246 TI - Time evolution of the extremely diluted Blume-Emery-Griffiths neural network. AB - A study of the time evolution and a stability analysis of the phases in the extremely diluted Blume-Emery-Griffiths neural network model are shown to yield new phase diagrams in which fluctuation retrieval may drive pattern retrieval. It is shown that saddle-point solutions associated with fluctuation overlaps slow down the flow of the network states towards the retrieval fixed points. A comparison of the performance with other three-state networks is also presented. PMID- 14754248 TI - Local scale invariance as dynamical space-time symmetry in phase-ordering kinetics. AB - The scaling of the spatiotemporal response of coarsening systems is studied through simulations of the two-dimensional (2D) and 3D Ising model with Glauber dynamics. The scaling functions are consistent with the prediction of local scale invariance, thereby suggesting the extension of dynamical scaling to a space-time dynamical symmetry. PMID- 14754247 TI - Influence of nonexcitable cells on spiral breakup in two-dimensional and three dimensional excitable media. AB - We study spiral wave dynamics in the presence of nonexcitable cells in two dimensional (2D) and three-dimensional (3D) excitable media, described by the Aliev-Panfilov model. We find that increasing the percentage of randomly distributed nonexcitable cells can prevent the breaking up of a spiral wave into a complex spatiotemporal pattern. We show that this effect is more pronounced in 2D than 3D excitable media. We explain the observed 2D vs 3D differences by a different dependence of the period and diastolic interval of the spiral wave on the percentage of nonexcitable cells in 2D and 3D excitable media. PMID- 14754249 TI - From multiplicative noise to directed percolation in wetting transitions. AB - A simple one-dimensional microscopic model of the depinning transition of an interface from an attractive hard wall is introduced and investigated. Upon varying a control parameter, the critical behavior observed along the transition line changes from a directed-percolation type to a multiplicative-noise type. Numerical simulations allow for a quantitative study of the multicritical point separating the two regions. Mean-field arguments and the mapping on yet a simpler model provide some further insight on the overall scenario. PMID- 14754250 TI - Self-similar community structure in a network of human interactions. AB - We propose a procedure for analyzing and characterizing complex networks. We apply this to the social network as constructed from email communications within a medium sized university with about 1700 employees. Email networks provide an accurate and nonintrusive description of the flow of information within human organizations. Our results reveal the self-organization of the network into a state where the distribution of community sizes is self-similar. This suggests that a universal mechanism, responsible for emergence of scaling in other self organized complex systems, as, for instance, river networks, could also be the underlying driving force in the formation and evolution of social networks. PMID- 14754251 TI - Measuring the fluctuation-dissipation ratio in glassy systems with no perturbing field. AB - A method is presented for measuring the integrated linear response in Ising spin system without applying any perturbing field. Large-scale simulations are performed in order to show how the method works. Very precise measurements of the fluctuation-dissipation ratio are presented for three different Ising models: the two-dimensional ferromagnetic model, the mean-field diluted three-spin model, and the three-dimensional Edwards-Anderson model. PMID- 14754252 TI - Synchronization and symmetry-breaking bifurcations in constructive networks of coupled chaotic oscillators. AB - The spatiotemporal dynamics of networks based on a ring of coupled oscillators with regular shortcuts beyond the nearest-neighbor couplings is studied by using master stability equations and numerical simulations. The generic criterion for dynamic synchronization has been extended to arbitrary network topologies with zero row-sum. The symmetry-breaking oscillation patterns that resulted from the Hopf bifurcation from synchronous states are analyzed by the symmetry group theory. PMID- 14754253 TI - Asymptotic scaling of the diffusion coefficient of fluctuating "pulled" fronts. AB - We present a (heuristic) theoretical derivation for the scaling of the diffusion coefficient D(f) for fluctuating "pulled" fronts. In agreement with earlier numerical simulations, we find that as N--> infinity, D(f) approaches zero as 1/ln(3) N, where N is the average number of particles per correlation volume in the stable phase of the front. This behavior of D(f) stems from the shape fluctuations at the very tip of the front, and is independent of the microscopic model. PMID- 14754254 TI - Resonant emergence of global and local spatiotemporal order in a nonlinear field model. AB - We investigate the nonequilibrium evolution of a scalar field in (2+1) dimensions. The field is set in a double-well potential in contact (open) or not (closed) with a heat bath. For closed systems, we observe the synchronized emergence of coherent spatiotemporal configurations, identified with oscillons. This initial global ordering degenerates into localized order until all oscillons disappear. We show that the synchronization is driven by resonant parametric oscillations of the field's zero mode and that local ordering is only possible outside equipartition. None of these orderings occur for open systems. PMID- 14754255 TI - Asymmetrical singularities in real-world signals. AB - We generalize the wavelet transform modulus maxima approach in order to analyze positive and negative changes separately and show different singularity spectra depending on the direction of changes in (i) human heartbeat interval data during sympathetic blockade, (ii) time series of daytime human physical activity of healthy individuals (but not of patients with debilitating fatigue), and (iii) daily stock price records of the Nikkei 225 in the period 1990-2002--but not of the S&P 500. We conclude that the analysis of asymmetrical singularities provides deeper insights into the underlying complexity of real-world signals that can greatly enhance our understanding of the mechanisms determining the systems' dynamics. PMID- 14754256 TI - Low-rank perturbations and the spectral statistics of pseudointegrable billiards. AB - We present an efficient method to solve Schrodinger's equation for perturbations of low rank. The method is ideally suited for systems with short range interactions or quantum billiards. It involves a secular equation of low dimension, which directly returns the level counting function. For illustration, we calculate the number variance for two pseudointegrable quantum billiards: the barrier billiard and a right triangle billiard. In this way, we obtain precise estimates for the level compressibility in the semiclassical (high energy) limit. In both cases, our results confirm recent theoretical predictions, based on periodic orbit summation, disregarding diffractive orbits. PMID- 14754257 TI - Scaling evolution in shock-induced transition to turbulence. AB - In this experimental study, a column of heavy gas (SF6) surrounded by light gas (air) is accelerated by a planar Mach 1.2 shock. Richtmyer-Meshkov instability on the initially diffuse air-SF6 interface determines the repeatable large-scale vortex dynamics of the system after the shock passage. Subsequently secondary instabilities form, with the system eventually transitioning to turbulence. We present highly resolved measurements of two components of the instantaneous velocity fields. With these measurements, we investigate the evolution of velocity statistics over a substantial range of scales in terms of structure functions. The latter evolve to exhibit late-time behavior consistent with the Kolmogorov scaling law for fully developed turbulence, despite the transitional character, anisotropy, and inhomogeneity of our flow. Ensemble averaging and comparison with instantaneous results reveal a trend towards the same scaling manifested much earlier by the structure functions of the fluctuating velocity components. PMID- 14754258 TI - Reciprocity theorem and perturbation theory for photonic crystal waveguides. AB - Starting from Maxwell's equations we derive a reciprocity theorem for photonic crystal waveguides. A set of strongly coupled discrete equations results, which can be applied to the simulation of perturbed photonic crystal waveguides. As an example we analytically study the influence of the dispersion of a two level system on the band structure of a photonic crystal waveguide. In particular, the formation of polariton gaps is discussed. PMID- 14754259 TI - Resonant and antiresonant frequency dependence of the effective parameters of metamaterials. AB - We present a numerical study of the electromagnetic response of the metamaterial elements that are used to construct materials with negative refractive index. For an array of split ring resonators (SRR) we find that the resonant behavior of the effective magnetic permeability is accompanied by an antiresonant behavior of the effective permittivity. In addition, the imaginary parts of the effective permittivity and permeability are opposite in sign. We also observe an identical resonant versus antiresonant frequency dependence of the effective materials parameters for a periodic array of thin metallic wires with cuts placed periodically along the length of the wire, with roles of the permittivity and permeability reversed from the SRR case. We show in a simple manner that the finite unit cell size is responsible for the antiresonant behavior. PMID- 14754260 TI - Spatial solitons in type II quasiphase-matched slab waveguides. AB - The existence and dynamics of one-dimensional spatial solitons formed upon propagation in quasiphase-matched gratings, through three-wave parametric interaction, is analyzed. We study the general case in which the grating exhibits a periodic modulation of both the refractive index and the second-order susceptibility. It is demonstrated that for negative effective wave vector mismatch the induced third-order nonlinearities increase the domain of soliton instability. Finally, the dependence of the efficiency of the second harmonic generation process in the soliton regime, on the parameters of the grating, is discussed. PMID- 14754261 TI - Vectorial Swift-Hohenberg equation for transverse laser patterns. AB - The consequences of introducing the polarization degree of freedom of the light are studied for the transverse patterns of a laser with detuning equal to zero. We deduce the vectorial Swift-Hohenberg amplitude equation from the corresponding Maxwell-Bloch equations. The vectorial character of the equation introduces modifications in the stability of traveling waves and new types of localized structures. PMID- 14754262 TI - Stabilization of one-dimensional periodic waves by saturation of the nonlinear response. AB - We address the properties of (1+1)-dimensional periodic waves in conservative saturable cubic nonlinear media and discover that cnoidal- and snoidal-type waves are completely stable within a broad range of parameters. The existence of stability bands is in sharp contrast with the previously known properties of periodic waves in self-focusing Kerr nonlinear media. We also found that in self defocusing media instability bands occur, again in contrast to the case of Kerr media. PMID- 14754263 TI - Nonequilibrium phase transition in a model for the propagation of innovations among economic agents. AB - We characterize the different morphological phases that occur in a simple one dimensional model of propagation of innovations among economic agents [X. Guardiola et al., Phys. Rev E 66, 026121 (2002)]. We show that the model can be regarded as a nonequilibrium surface growth model. This allows us to demonstrate the presence of a continuous roughening transition between a flat (system size independent fluctuations) and a rough phase (system size dependent fluctuations). Finite-size scaling studies at the transition strongly suggest that the dynamic critical transition does not belong to directed percolation and, in fact, critical exponents do not seem to fit in any of the known universality classes of nonequilibrium phase transitions. Finally, we present an explanation for the occurrence of the roughening transition and argue that avalanche driven dynamics is responsible for the novel critical behavior. PMID- 14754264 TI - Scaling laws for the movement of people between locations in a large city. AB - Large scale simulations of the movements of people in a "virtual" city and their analyses are used to generate insights into understanding the dynamic processes that depend on the interactions between people. Models, based on these interactions, can be used in optimizing traffic flow, slowing the spread of infectious diseases, or predicting the change in cell phone usage in a disaster. We analyzed cumulative and aggregated data generated from the simulated movements of 1.6 x 10(6) individuals in a computer (pseudo-agent-based) model during a typical day in Portland, Oregon. This city is mapped into a graph with 181,206 nodes representing physical locations such as buildings. Connecting edges model individual's flow between nodes. Edge weights are constructed from the daily traffic of individuals moving between locations. The number of edges leaving a node (out-degree), the edge weights (out-traffic), and the edge weights per location (total out-traffic) are fitted well by power-law distributions. The power-law distributions also fit subgraphs based on work, school, and social/recreational activities. The resulting weighted graph is a "small world" and has scaling laws consistent with an underlying hierarchical structure. We also explore the time evolution of the largest connected component and the distribution of the component sizes. We observe a strong linear correlation between the out-degree and total out-traffic distributions and significant levels of clustering. We discuss how these network features can be used to characterize social networks and their relationship to dynamic processes. PMID- 14754265 TI - Nanoscopic liquid bridges exposed to a torsional strain. AB - In this paper we investigate the response to a torsional strain of a molecularly thin film of spherically symmetric molecules confined to a chemically heterogeneous slit pore by means of Monte Carlo simulations in the grand canonical ensemble. The slit pore comprises two identical plane-parallel solid substrates, the fluid-substrate interaction is purely repulsive except for elliptic regions attracting fluid molecules. Under favorable thermodynamic conditions the confined film consists of fluid bridges where the molecules are preferentially adsorbed by the attractive elliptic regions, and span the gap between the opposite substrate surfaces. By rotating the upper substrate while holding the lower one in position, bridge phases can be exposed to a torsional strain 0< or =theta< or =pi/2 and the associated torsional stress T(theta) of the (fluidic) bridge phases can be calculated from molecular expressions. The obtained stress curve T(theta)(theta) is qualitatively similar to the one characteristic of sheared confined films: as the torsion strain increases, T(theta) rises to a maximum (yield point) and then decays monotonically to zero. By changing the ellipses' aspect ratio while keeping their area constant, we also investigate the influence of the attractive elliptic patterns' shape on T(theta)(theta). PMID- 14754266 TI - Properties of a random attachment growing network. AB - In this study we introduce and analyze the statistical structural properties of a model of growing networks which may be relevant to social networks. At each step a new node is added which selects k possible partners from the existing network and joins them with probability delta by undirected edges. The "activity" of the node ends here; it will get new partners only if it is selected by a newcomer. The model produces an infinite-order phase transition when a giant component appears at a specific value of delta, which depends on k. The average component size is discontinuous at the transition. In contrast, the network behaves significantly different for k=1. There is no giant component formed for any delta and thus in this sense there is no phase transition. However, the average component size diverges for delta> or =1/2. PMID- 14754267 TI - Phase transitions of the anisotropic Ashkin-Teller model on a family of diamond type hierarchical lattices. AB - The phase transitions of the anisotropic Ashkin-Teller model on a family of diamond-type hierarchical lattices is studied by means of the transfer-matrix method and the real-space renormalization-group transformation. We find that the phase diagram, for the ferromagnetic case, consists of five phases, i.e., the fully disordered paramagnetic phase P, the fully ordered ferromagnetic phase F, and three partially ordered ferromagnetic phases F(s), F(sigma), and F(s sigma), as well as ten nontrivial fixed points. The correlation length critical exponents and the crossover exponents are also calculated. In addition, we also investigate the variations of the critical exponents with the fractal dimension d(f), the number of branches m, and the number of bonds per branch b of the generator of the family of diamond-type hierarchical lattices. Finally we give a brief discussion about universality. PMID- 14754268 TI - Where the linearized Poisson-Boltzmann cell model fails: the planar case as a prototype study. AB - The linearized Poisson-Boltzmann (PB) approximation is investigated for the classical problem of two infinite, uniformly charged planes in electrochemical equilibrium with an infinite monovalent salt reservoir. At the nonlinear level, we obtain an explicit expression of the associated electrostatic contribution to the semi-grand-canonical potential. The linearized osmotic-pressure difference between the interplane region and the salt reservoir becomes negative in the low temperature, large-separation, or high-surface charge limits, in disagreement with the exact (at mean-field level) nonlinear PB solution. We show that these artifacts--although thermodynamically consistent with quadratic expansions of the nonlinear functional--can be traced back to the nonfulfillment of the underlying assumptions of the linearization. Explicit comparison between the analytical expressions of the exact nonlinear solution and the corresponding linearized equations allows us to show that the linearized results are asymptotically exact in the weak-coupling and counterionic ideal-gas limits, but always fail otherwise, predicting negative osmotic-pressure differences. By taking appropriate limits of the full nonlinear PB solution, we provide asymptotic expressions for the semi-grand-canonical potential and the osmotic-pressure difference that involve only elementary functions, which cover the complementary region where the linearized theory breaks down. PMID- 14754269 TI - Scaling laws for critical manifolds in polycrystalline materials. AB - We study the surfaces of lowest energy through model polycrystalline materials in two and three dimensions. When the grain boundaries are sufficiently weak, these critical manifolds (CM's) lie entirely on grain boundaries, while when the grain boundaries are strong, cleavage occurs. A scaling theory for the intergranular to transgranular transition of CM's is developed. The key parameters are the average grain size g, the ratio of grain boundary to the grain interior energy, epsilon, and the sample size L. The key result is that a critical length scale exists, L(c)(g,epsilon), so that on short length scales lL(c), the critical manifold is rough. We develop a scaling theory for L(c) and find that in two dimensions L(c) approximately gx(y(2)), while in three dimensions L(c) approximately g exp(bx(y(3))), where x=epsilon/(1-epsilon) and b is a constant. Data from realistic polycrystalline grain structures are used to test the scaling theory. The exact lowest energy surface through model grain structures is found using a mapping to the minimum cut/maximum-flow problem in computer science. As a function of grain-boundary energy, we observe the crossover from grain-boundary rupture to mixed mode failure (a mixture of transgramular and intergranular modes) and finally cleavage and that the two-dimensional data are consistent with y(2) approximately 3.0+/ 0.3, while the three-dimensional data are more difficult to analyze, but are consistent with y(3) approximately 3.5+/-1.0. PMID- 14754271 TI - Analysis of stochastic resonances. AB - We investigate the one-dimensional diffusion of a particle in a piecewise linear potential superimposed with a step of a harmonically modulated height. Employing the matching conditions, we solve the corresponding Fokker-Planck equation and we analyze nonlinear features of the particle's mean position as a function of time. We present detailed results in two physically relevant cases. First, we take the unperturbed potential as a symmetrical up-oriented tip, which is placed between two reflecting boundaries and we add the jump at the tip coordinate. The setting yields resonance-like behavior of the stationary-response amplitude. Second, if the discontinuity at origin is combined with the constant force in the symmetrical region between the boundaries, the stationary response displays a time-independent shift against the potential slope. The driving-induced force exhibits a resonance-like behavior both with respect to the diffusion constant and the slope of the unperturbed potential. PMID- 14754270 TI - Influence of external information in the minority game. AB - The influence of a fixed number of agents with the same fixed behavior on the dynamics of the minority game is studied. Alternatively, the system studied can be considered the minority game with a change in the comfort threshold away from half filling. Agents in the frustrated, nonergodic phase tend to overreact to the information provided by the fixed agents, leading not only to large fluctuations, but to deviations of the average occupancies from their optimal values. Agents that discount their impact on the market, or that use individual strategies reach equilibrium states, which, unlike in the absence of the external information provided by the fixed agents, do not give the highest payoff to the collective. PMID- 14754272 TI - Thermodynamics of electrolytes on anisotropic lattices. AB - The phase behavior of ionic fluids on simple cubic and tetragonal (anisotropic) lattices has been studied by grand canonical Monte Carlo simulations. Systems with both the true lattice Coulombic potential and continuous-space 1/r electrostatic interactions have been investigated. At all degrees of anisotropy, only coexistence between a disordered low-density phase and an ordered high density phase with the structure similar to ionic crystal was found, in contrast to recent theoretical predictions. Tricritical parameters were determined to be increasing functions of anisotropy parameters which is consistent with theoretical calculations based on the Debye-Huckel approach. At large anisotropies a two-dimensional-like behavior is observed, from which we estimated the dimensionless tricritical temperature and density for the two-dimensional square lattice electrolyte to be T*(tri)=0.14 and rho*(tri)=0.70. PMID- 14754273 TI - Monodisperse model suitable to study the glass transition. AB - We study the properties of a monodisperse lattice glass model with a simple geometrical interpretation, which reproduces many features of glass forming liquids, such as the cage effect, vanishing diffusivity, and the presence of two time scales in relaxation functions. The model has a crystalline ground state at high density, but has no tendency to crystallize when quenched, even at extremely low cooling rates, which makes it suitable for the study of the glass transition. We study the model in mean field on random regular graphs, finding a scenario analogous to p-spin models. PMID- 14754274 TI - Quantum master equation for a system influencing its environment. AB - A perturbative quantum master equation is derived for a system interacting with its environment, which is more general than the ones derived before. Our master equation takes into account the effect of the energy exchanges between the system and the environment and the conservation of energy in the finite total system. This master equation describes relaxation mechanisms in isolated nanoscopic quantum systems. In its most general form, this equation is non-Markovian and a Markovian version of it rules the long-time relaxation. We show that our equation reduces to the Redfield equation in the limit where the energy of the system does not affect the density of state of its environment. This master equation and the Redfield one are applied to a spin-environment model defined in terms of random matrices and compared with the solutions of the exact von Neumann equation. The comparison proves the necessity to allow energy exchange between the subsystem and the environment in order to correctly describe the relaxation in an isolated nanoscopic total system. PMID- 14754275 TI - Spin relaxation in a complex environment. AB - We report the study of a model of a two-level system interacting in a nondiagonal way with a complex environment described by Gaussian orthogonal random matrices (GORM). The effect of the interaction on the total spectrum and its consequences on the dynamics of the two-level system is analyzed. We show the existence of a critical value of the interaction, depending on the mean level spacing of the environment, above which the dynamics is self-averaging and closely obey a master equation for the time evolution of the observables of the two-level system. Analytic results are also obtained in the strong coupling regimes. We finally study the equilibrium values of the two-level system population and show under which condition it thermalizes to the environment temperature. PMID- 14754276 TI - Large-scale emergent properties of an autocatalytic reaction-diffusion model subject to noise. AB - The nonequilibrium dynamic fluctuations of a stochastic version of the Gray-Scott (GS) model are studied analytically in leading order in perturbation theory by means of the dynamic renormalization group. There is an attracting stable fixed point at one-loop order, and the asymptotic scaling of the correlation functions is predicted for both spatial and temporally correlated noise sources. New effective three-body reaction terms, not present in the original GS model, are induced by the combined interplay of the fluctuations and nonlinearities. PMID- 14754277 TI - Crossover between special and ordinary transitions in random semi-infinite Ising like systems. AB - We consider the crossover behavior between special and ordinary surface transitions in three-dimensional semi-infinite Ising-like systems with random quenched bulk disorder. We calculate the surface crossover critical exponent Phi, the critical exponents of the layer alpha(1), and local specific heats alpha(11) by applying the field theoretic approach directly in three spatial dimensions (d=3) up to the two-loop approximation. The numerical estimates of the resulting two-loop series expansions for the surface critical exponents are computed by means of Pade and Pade-Borel resummation techniques. We find that Phi, alpha(1), alpha(11) obtained in the present paper are different from their counterparts of pure Ising systems. The obtained results support the idea that in a system with random quenched bulk disorder the plane boundary is characterized by a new set of critical exponents. PMID- 14754278 TI - Alternative model of the Antonov problem. AB - Astrophysical systems will never be in a real thermodynamic equilibrium: they undergo an evaporation process due to the fact that the gravity is not able to confine the particles. Ordinarily, this difficulty is overcome by enclosing the system in a rigid container which avoids the evaporation. We propose an energetic prescription which is able to confine the particles, leading in this way to an alternative version of the Antonov isothermal model which unifies the well-known isothermal and polytropic profiles. Besides the main features of the isothermal sphere model (the existence of the gravitational collapse and the energetic region with a negative specific heat), this alternative model has the advantage that the system size naturally appears as a consequence of the particles' evaporation. PMID- 14754279 TI - Properties of chord length distributions across ordered and disordered packing of hard disks. AB - Chord length distributions across packings--random or not--of equal hard disks have a universal divergence which is proportional to n(c)/square root of l for small chords, where n(c) is the mean number of contacts. A similar behavior is derived for a population of polydisperse disks. Monte Carlo simulations across various kinds of regular and random packings are in full agreement with the theoretical predictions. PMID- 14754280 TI - Friction in the zero sliding velocity limit. AB - Using an adiabatic approximation method, which searches for Tomlinson model-like instabilities for a simple but still realistic model for two crystalline surfaces, with mobile molecules present at the interface, sliding relative to each other, we are able to account for the virtually universal occurrence of "dry friction" at zero temperature. A modified version of this method allows us to calculate the kinetic friction at nonzero temperature as well. We have also considered the static friction, and have demonstrated that the model is able to account for static friction being larger than kinetic friction. PMID- 14754282 TI - Density functional theory for nearest-neighbor exclusion lattice gases in two and three dimensions. AB - To speak about fundamental measure theory obliges us to mention dimensional crossover. This feature, inherent to the systems themselves, was incorporated in the theory almost from the beginning. Although at first it was thought to be a consistency check for the theory, it rapidly became its fundamental pillar, thus becoming the only density functional theory which possesses such a property. It is straightforward that dimensional crossover connects, for instance, the parallel hard cube system (three dimensional) with that of squares (two dimensional) and rods (one dimensional). We show here that there are many more connections which can be established in this way. Through them we deduce from the functional for parallel hard (hyper)cubes in the simple (hyper)cubic lattice the corresponding functionals for the nearest-neighbor exclusion lattice gases in the square, triangular, simple cubic, face-centered-cubic, and body-centered-cubic lattices. As an application, the bulk phase diagram for all these systems is obtained. PMID- 14754281 TI - Some asymptotic properties of duplication graphs. AB - Duplication graphs are graphs that grow by duplication of existing vertices, and are important models of biological networks, including protein-protein interaction networks and gene regulatory networks. Three models of graph growth are studied: pure duplication growth, and two two-parameter models in which duplication forms one element of the growth dynamics. A power-law degree distribution is found to emerge in all three models. However, the parameter space of the latter two models is characterized by a range of parameter values for which duplication is the predominant mechanism of graph growth. For parameter values that lie in this "duplication-dominated" regime, it is shown that the degree distribution either approaches zero asymptotically, or approaches a nonzero power-law degree distribution very slowly. In either case, the approach to the true asymptotic degree distribution is characterized by a dependence of the scaling exponent on properties of the initial degree distribution. It is therefore conjectured that duplication-dominated, scale-free networks may contain identifiable remnants of their early structure. This feature is inherited from the idealized model of pure duplication growth, for which the exact finite-size degree distribution is found and its asymptotic properties studied. PMID- 14754283 TI - Crossover behavior in a communication network. AB - We address the problem of message transfer in a communication network. The network consists of nodes and links, with the nodes lying on a two-dimensional lattice. Each node has connections with its nearest neighbors, whereas some special nodes, which are designated as hubs, have connections to all the sites within a certain area of influence. The degree distribution for this network is bimodal in nature and has finite variance. The distribution of travel times between two sites situated at a fixed distance on this lattice shows fat-fractal behavior as a function of hub density. If extra assortative connections are now introduced between the hubs so that each hub is connected to two or three other hubs, the distribution crosses over to power-law behavior. Crossover behavior is also seen if end-to-end short cuts are introduced between hubs whose areas of influence overlap, but this is much milder in nature. In yet another information transmission process, namely, the spread of infection on the network with assortative connections, we again observed crossover behavior of another type, viz., from one power law to another for the threshold values of disease transmission probability. Our results are relevant for the understanding of the role of network topology in information spread processes. PMID- 14754284 TI - General theory of nonlinear flow-distributed oscillations. AB - We outline a general theory for the analysis of flow-distributed standing and traveling wave patterns in one-dimensional, open flows of oscillatory chemical media, emphasizing features that are generic to a variety of kinetic models. We draw particular attention to the cases far from a Hopf bifurcation and far from the so-called kinematic or zero-diffusion limit. We introduce a nonlinear formalism for both traveling and stationary waves and show that the wave forms and their amplitudes depend on a single reduced transport parameter that quantifies the departure from the kinematic limit. The nonlinear formalism can be applied to systems with more complex types of bifurcations (canards, period doublings, etc.). PMID- 14754285 TI - Continuum traffic model with the consideration of two delay time scales. AB - This paper presents a continuum traffic model. The derivation of this model is based upon the assumption that the stream velocity u reaches the equilibrium velocity u(e) within the relaxation time T, while the equilibrium velocity u(e) is adjusted to be attained through the driver's reaction time t(r). It is also assumed that the former delay time scale is greater than the latter. A motion equation with nonconstant propagation velocity of a disturbance in traffic flow is derived that can reflect the anisotropy of disturbance propagation in real traffic, unlike some other higher-order continuum models. It indicates that in our model the undesirable "wrong-way travel" phenomenon and gas-like behavior have been eliminated. The formation and diffusion of traffic shock can be better simulated. PMID- 14754286 TI - Stochastically evolving networks. AB - We discuss a class of models for the evolution of networks in which new nodes are recruited into the network at random times, and links between existing nodes that are not yet directly connected may also form at random times. The class contains both models that produce "small-world" networks and less tightly linked models. We produce both trees, appropriate in certain biological applications, and networks in which closed loops can appear, which model communication networks and networks of human sexual interactions. One of our models is closely related to random recursive trees, and some exact results known in that context can be exploited. The other models are more subtle and difficult to analyze. Our analysis includes a number of exact results for moments, correlations, and distributions of coordination number and network size. We report simulations and also discuss some mean-field approximations. If the system has evolved for a long time and the state of a random node (which thus has a random age) is observed, power-law distributions for properties of the system arise in some of these models. PMID- 14754287 TI - Microcanonical Monte Carlo simulation of lattice gas models. AB - We introduce a true microcanonical ensemble appropriate for Monte Carlo simulations of lattice gas models in which both the energy and the number of particles are held fixed. We also deduce formulas that allow us to do the calculation of the intensive quantities: temperature and chemical potential. The numerical results coming from Monte Carlo simulations defined according to this microcanonical ensemble compare well with results coming from the ordinary canonical ensemble. PMID- 14754288 TI - Structure of equations of macrophysics. AB - The first and the second laws of thermodynamics form the constraints on the equations describing macrophysical phenomena. It is argued in this paper that there are some additional universal constraints. These constraints are caused by the Hamiltonian structure of microequations. Previously one feature of micromotion, its reversibility, was used by Onsager to explain the observed reciprocity relations. Hamiltonian structure is richer than reversibility and yields richer consequences. Some of these consequences are a nonlinear version of Onsager's relations, Hamiltonian structure of reversible equations of macrophysics, and quasi-Hamiltonian structure of irreversible equations. PMID- 14754289 TI - Nonequilibrium critical dynamics of the triangular antiferromagnetic Ising model. AB - We investigate the nonequilibrium critical dynamics of antiferromagnetic Ising model on a two-dimensional triangular lattice via dynamic Monte Carlo simulation employing spin-flip kinetics. Macroscopic degeneracy of the ground state originating from geometric frustration fundamentally affects the nonequilibrium dynamics of the system. In particular, the defects and the loose spins (whose flip costs no energy) play key roles in the dynamics. The long-time evolution is characterized by a critical dynamic scaling with a growing length scale xi(t). With random initial configurations, xi(t) exhibits a subdiffusive growth in time, xi(t) approximately t(1/z) with 1/z approximately 0.43, while xi(t) shows a diffusive growth with z=2 for the relaxation within the dominant sector of the ground-state manifold. The nonequilibrium critical dynamics therefore exhibits an interesting initial-state dependence. Persistence and the two-time temporal properties are also discussed. PMID- 14754290 TI - Quasiequilibrium during aging of the two-dimensional Edwards-Anderson model. AB - We test the quasiequilibrium picture of the aging dynamics--strictly valid in the asymptotic dynamical regime of aging systems--in the preasymptotic aging regime of the two-dimensional Edwards-Anderson spin glass model. We compare the fluctuation-dissipation characteristic for spin autocorrelation function and response with a corresponding one obtained for a suitably defined correlation function and its conjugated response. In agreement with the quasiequilibrium picture we find that after a short transient the two corresponding fluctuation dissipation ratios (FDR's) coincide at equal times. Moreover we show that, as it happens for the usual FDR, the dynamic FDR at finite time coincides with the static one at finite size. PMID- 14754291 TI - Ballistic localization in quasi-one-dimensional waveguides with rough surfaces. AB - Structure of eigenstates in a periodic quasi-one-dimensional waveguide with a rough surface is studied both analytically and numerically. We have found a large number of "regular" eigenstates for any high energy. They result in a very slow convergence to the classical limit in which the eigenstates are expected to be completely ergodic. As a consequence, localization properties of eigenstates originated from unperturbed transverse channels with low indexes are strongly localized (delocalized) in the momentum (coordinate) representation. These eigenstates were found to have a quite unexpected form that manifests a kind of "repulsion" from the rough surface. Our results indicate that standard statistical approaches for ballistic localization in such waveguides seem to be inappropriate. PMID- 14754292 TI - Bivariate surrogate techniques: necessity, strengths, and caveats. AB - The concept of surrogates allows testing results from time series analysis against specified null hypotheses. In application to bivariate model dynamics we here compare different types of surrogates, each designed to test against a different null hypothesis, e.g., an underlying bivariate linear stochastic process. Two measures that aim at a characterization of interdependence between nonlinear deterministic dynamics were used as discriminating statistics. We analyze eight different stochastic and deterministic models not only to demonstrate the power of the surrogates, but also to reveal some pitfalls and limitations. PMID- 14754293 TI - Dynamical origin for the occurrence of asynchronous hyperchaos and chaos via blowout bifurcations. AB - We investigate the dynamical origin for the occurrence of asynchronous hyperchaos and chaos via blowout bifurcations in coupled chaotic systems. An asynchronous hyperchaotic or chaotic attractor with a positive or negative second Lyapunov exponent appears through a blowout bifurcation. It is found that the sign of the second Lyapunov exponent of the newly born asynchronous attractor, exhibiting on off intermittency, is determined through competition between its laminar and bursting components. When the "strength" (i.e., a weighted second Lyapunov exponent) of the bursting component is larger (smaller) than that of the laminar component, an asynchronous hyperchaotic (chaotic) attractor appears. PMID- 14754294 TI - Pattern formation at the bicritical point of the Faraday instability. AB - We present measurements on parametrically driven surface waves (Faraday waves) performed in the vicinity of a bicritical point in parameter space, where modes with harmonic and subharmonic time dependence interact. The primary patterns are squares in the subharmonic and hexagons in the harmonic regime. If the primary instability is harmonic we observe a hysteretic secondary transition from hexagons to squares without a perceptible variation of the fundamental wavelength. The transition is understood in terms of a set of coupled Landau equations and related to other canonical examples of phase transitions in nonlinear dissipative systems. Moreover, the subharmonic-harmonic mode competition gives rise to a variety of new superlattice states. These structures are interpreted as mediator modes involved in the transition between patterns of fourfold and sixfold rotational symmetry. PMID- 14754295 TI - External cavity modes of semiconductor lasers with phase-conjugate feedback. AB - External cavity modes (ECMs) of a semiconductor laser with phase-conjugate feedback are defined as time-periodic pulsating intensity solutions exhibiting a frequency close to an integer multiple of the external cavity frequency. As the feedback rate progressively increases from zero, they sequentially appear as stable attractors in the bifurcation diagram. We construct a simple analytical approximation of these pulsating intensity solutions and determine their frequencies. We show that branches of ECMs are isolated. Finally, the validity of our approximation is tested by comparing numerical bifurcation diagrams obtained by simulation and continuation techniques with our analytical results. PMID- 14754296 TI - Noise-controlled oscillations and their bifurcations in coupled phase oscillators. AB - We derive in Gaussian approximation dynamical equations for the first two cumulants of the mean field fluctuations in a system of globally coupled stochastic phase oscillators. In these equations the intensity of noise serves as an explicit control parameter. Its variation generates transitions between three dynamical regimes: (i) stationary, (ii) rotatory and (iii) locally oscillatory (breathing). The latter regime has previously not been reported in studies of globally coupled noisy phase oscillators. Our detailed bifurcation analysis is supported by numerical simulations of an ensemble of coupled stochastic phase oscillators. Similar regimes are also found in simulations of globally coupled stochastic FitzHugh-Nagumo elements. PMID- 14754297 TI - Vortex formation of coherent waves in nonseparable mesoscopic billiards. AB - We analytically study the vortex formation associated with the classical periodic orbits in a nonseparable quantum billiard. The influence of vortex formation on the quantum flux in the mesoscopic region is clearly demonstrated. Remarkably, the high-order vortex structures in an equilateral triangle billiard display the triangular, swirl, and kagome vortex lattices. The relationship between various vortex lattices is found to rely on the relative phase factor in the coherent state. PMID- 14754298 TI - Self-stabilization of high-frequency oscillations in semiconductor superlattices by time-delay autosynchronization. AB - We present a scheme to stabilize high-frequency domain oscillations in semiconductor superlattices by a time-delayed feedback loop. Applying concepts from chaos control theory we propose to control the spatiotemporal dynamics of fronts of accumulation and depletion layers which are generated at the emitter and may collide and annihilate during their transit, and thereby suppress chaos. The proposed method only requires the feedback of internal global electrical variables, viz., current and voltage, which makes the practical implementation very easy. PMID- 14754299 TI - Competition of synchronization domains in arrays of chaotic homoclinic systems. AB - We investigate the response of an open chain of bidirectionally coupled chaotic homoclinic systems to external periodic stimuli. When one end of the chain is driven by a periodic signal, the system propagates a phase synchronization state in a certain range of coupling strengths and external frequencies. When two simultaneous forcings are applied at different points of the array, a rich phenomenology of stable competitive states is observed, including temporal alternation and spatial coexistence of synchronization domains. PMID- 14754300 TI - Relaxation and diffusion in a globally coupled Hamiltonian system. AB - The relation between relaxation and diffusion is investigated in a Hamiltonian system of globally coupled rotators. Diffusion is anomalous if and only if the system is going towards equilibrium. The anomaly in diffusion is not anomalous diffusion taking a power-type function, but is a transient anomaly due to nonstationarity. For a certain type of initial condition, in quasistationary states, diffusion can be explained by a stretched exponential correlation function, whose stretching exponent is almost constant and correlation time is linear as functions of degrees of freedom. The full time evolution is characterized by varying stretching exponent and correlation time. PMID- 14754301 TI - Experimental demonstration of attractor annihilation in a multistable fiber laser. AB - We report on the experimental open-loop control of generalized multistability in a system with coexisting attractors. The experimental system is an erbium-doped fiber laser with pump modulation of the diode laser. We demonstrate that additional weak harmonic modulation of the diode current annihilates one or two stable limit cycles in the laser. The ability of the method to select a desired state is illustrated through a codimension-two bifurcation diagram in the parameter space of the frequency and amplitude of the control modulation. We identify main resonances on the bifurcation lines (annihilation curves) and evaluate conditions for attractor annihilation. PMID- 14754302 TI - Controlling chaos in a Lorenz-like system using feedback. AB - We demonstrate that the dynamics of an autonomous chaotic laser can be controlled to a periodic or steady state under self-synchronization. In general, past the chaos threshold the dependence of the laser output on feedback applied to the pump is submerged in the Lorenz-like chaotic pulsation. However there exist specific feedback delays that stabilize the chaos to periodic behavior or even steady state. The range of control depends critically on the feedback delay time and amplitude. Our experimental results are compared with the complex Lorenz equations which show good agreement. PMID- 14754303 TI - Scaling properties of saddle-node bifurcations on fractal basin boundaries. AB - We analyze situations where a saddle-node bifurcation occurs on a fractal basin boundary. Specifically, we are interested in what happens when a system parameter is slowly swept in time through the bifurcation. Such situations are known to be indeterminate in the sense that it is difficult to predict the eventual fate of an orbit that tracks the prebifurcation node attractor as the system parameter is swept through the bifurcation. In this paper we investigate the scaling of (1) the fractal basin boundary of the static (i.e., unswept) system near the saddle node bifurcation, (2) the dependence of the orbit's final destination on the sweeping rate, (3) the dependence of the time it takes for an attractor to capture a swept orbit on the sweeping rate, and (4) the dependence of the final attractor capture probability on the noise level. With respect to noise, our main result is that the effect of noise scales with the 5/6 power of the parameter drift rate. Our approach is to first investigate all these issues using one dimensional map models. The simplification of treatment inherent in one dimension greatly facilitates analysis and numerical experiment, aiding us in obtaining the new results listed above. Following our one-dimensional investigations, we explain that these results can be applied to two-dimensional systems. We show, through numerical experiments on a periodically forced second-order differential equation example, that the scalings we have found also apply to systems that result in two-dimensional maps. PMID- 14754304 TI - Multiphase control of a nonlinear lattice. AB - Large amplitude, multiphase excitations of the periodic Toda lattice (n-gap solutions) are created and controlled by small forcing. The approach uses passage through an ensemble of resonances and subsequent multiphase self-locking of the system with adiabatic wave-like perturbations. The synchronization of each phase in the excited lattice proceeds from the weakly nonlinear stage, where the problem can be reduced to that for a number of independent, driven, one-degree-of freedom oscillatory systems. Due to this separability, the phase locking at this stage is robust, provided the amplitude of the corresponding forcing component exceeds a threshold, which scales as 3/4 power of the corresponding frequency chirp rate. The adiabatic synchronization continues into a fully nonlinear stage, as the driven lattice self-adjusts its state to remain in a persisting and stable multifrequency resonance with the driving perturbation. Thus, a complete control of the n-gap state becomes possible by slow variation of external parameters. PMID- 14754305 TI - Signatures of classical diffusion in quantum fluctuations of two-dimensional chaotic systems. AB - We consider a two-dimensional (2D) generalization of the standard kicked rotor and show that it is an excellent model for the study of universal features of 2D quantum systems with underlying diffusive classical dynamics. First we analyze the distribution of wave-function intensities and compare them with the predictions derived in the framework of diffusive disordered samples. Next, we turn the closed system into an open one by constructing a scattering matrix. The distribution of the resonance widths P(Gamma) and Wigner delay times P(tau(W)) are investigated. The forms of these distributions are obtained for different symmetry classes and the traces of classical diffusive dynamics are identified. Our theoretical arguments are supported by extensive numerical calculations. PMID- 14754306 TI - Traveling waves in rotating Rayleigh-Benard convection: analysis of modes and mean flow. AB - Numerical simulations of the Boussinesq equations with rotation for realistic no slip boundary conditions and a finite annular domain are presented. These simulations reproduce traveling waves observed experimentally. Traveling waves are studied near threshold by using the complex Ginzburg-Landau equation (CGLE): a mode analysis enables the CGLE coefficients to be determined. The CGLE coefficients are compared with previous experimental and theoretical results. Mean flows are also computed and found to be more significant as the Prandtl number decreases (from sigma=6.4 to sigma=1). In addition, the mean flow around the outer radius of the annulus appears to be correlated with the mean flow around the inner radius. PMID- 14754307 TI - Well-behaved dynamics in a dissipative nonideal periodically kicked rotator. AB - Well-behaved dynamical properties are found in a dissipative kicked rotator subjected to a periodic string of asymmetric pulses of finite amplitude and width. The stability boundaries of the equilibrium are determined to arbitrary approximation for trigonometric pulses by means of circular harmonic balance, and to first approximation for general elliptic pulses by means of an elliptic harmonic balance method. The bifurcation behavior at the stability boundaries is determined numerically. We show how the extension of the instability region of the equilibrium in pulse parameter space reaches a maximum as the pulse width is varied. We also characterize the dependence of the mean duration of the transients to the equilibrium on the pulse width. The evolution of the basins of attraction of chaotic attractors when solely the pulse width is varied is characterized numerically. Finally, we show that the order-chaos route when solely the width of the pulses is altered appears to be especially rich, including different types of crises. The mechanism underlying these reshaping induced crises is discussed with the aid of a two-dimensional map. PMID- 14754308 TI - Synchronization of hyperchaotic systems with delayed bidirectional coupling. AB - Synchronization of three bidirectionally coupled hyperchaotic systems is studied, using the Ikeda model as the hyperchaotic unit. The whole system is given by three delay-differential equations with two distinct time lags. Sufficient condition for the global stability of the manifold of exact synchronization is found analytically. Local stability of the synchronization manifold is studied by numerical computation of the transverse Lyapunov exponent and statistical properties of the orbits. PMID- 14754309 TI - Stationary core-annular flow through a horizontal pipe. AB - A theoretical investigation has been made of core-annular flow: the flow of a high-viscosity liquid core surrounded by a low-viscosity annular liquid layer through a horizontal pipe. Special attention is paid to the question how the buoyancy force on the core, caused by a possible density difference between the core and the annular layer, is counterbalanced. From earlier studies it is known that at the core surface ripples are present that have the shape of "bamboo" waves or "snake" waves. They generate pressure variations and secondary flows in the annular layer that can cause a net hydrodynamic force on the core. Using hydrodynamic-lubrication theory (assuming the core to be rigid) it has been shown that for snake waves the lubrication force can counterbalance the buoyancy force. For bamboo waves that is not the case. PMID- 14754310 TI - Response maxima in modulated turbulence. II. Numerical simulations. AB - Numerical simulations of fully developed turbulence driven by a modulated energy input rate or driving force are performed within two dynamical cascade models, the Gkedzer-Ohkitani-Yamada shell model and a reduced wave vector set approximation of the Navier-Stokes equation. The frequency behavior of the system response is studied and compared with predictions from a variable range mean field theory, which excludes turbulent fluctuations. In agreement with the mean field approach, we find a constant response amplitude for low driving frequencies and a 1/omega decay of the amplitude for high frequencies. In the mean-field theory, the finite cascade time scale had led to an oscillating behavior of the response amplitude as a function of the driving frequency. In the simulations of both models we observe the main maximum. The higher maxima and minima are completely washed out by fluctuations, though the statistical properties of the fluctuations are different in the two models. PMID- 14754311 TI - Particle image velocimetry measurement of the velocity field in turbulent thermal convection. AB - The spatial structure of the velocity field in turbulent Rayleigh-Benard convection in water has been measured using the particle image velocimetry technique, with the Rayleigh number Ra varying from 9 x 10(8) to 9 x 10(11) and the Prandtl number remaining approximately constant (Pr approximately 4). The study provides a direct confirmation that a rotatory mean wind indeed persists for the highest value of Ra reached in the experiment. The measurement reveals that the mean flow in the central region of the convection cell is of the shape of a coherent elliptical rotating core for Ra below 1 x 10(10). Above this Ra, the orientation of the elliptical core changes by a 90 degrees angle and an inner core rotating at a lower rate inside the original bulk core emerges. It is further found that the rotation frequencies of the inner core and the outer shell have distinct scalings with Ra; the scaling exponent for the outer-shell is 0.5 and it is 0.4 for the inner core. From the measured rms and skewness distributions of the velocity field, we find that velocity fluctuations at the cell center are neither homogeneous nor isotropic. The turbulent energy production fields further reveal that the mean wind is not driven by turbulent fluctuations associated with Reynolds stress. PMID- 14754312 TI - Importance of the Darrieus-Landau instability for strongly corrugated turbulent flames. AB - The renormalization ideas of self-similar dynamics of a strongly turbulent flame front are applied to the case of a flame with realistically large thermal expansion of the burning matter. In that case a flame front is corrugated both by the external turbulence and by the intrinsic flame instability (the Darrieus Landau instability). The analytical formulas for the velocity of flame propagation are obtained. It is demonstrated that the flame instability is of principal importance when the maximal hydrodynamic length scale is much larger than the cutoff wavelength of the instability, provided the turbulent intensity is not too high. PMID- 14754313 TI - Self-similarity of two flows induced by instabilities. AB - The implications of full self-similarity of the Rayleigh-Taylor mixing layer and the Kelvin-Helmholtz shear layer are examined using a simplified group-theoretic analysis. The constraints on the behavior and evolution of these layers imposed by rigorous self-similarity are identified, and equations are constructed for the growth rate of these layers based on a total energy balance. This analysis does not prove that such flows will become self-similar. Rather, the analysis demonstrates the behaviors that would arise if these flows were to become fully self-similar. PMID- 14754314 TI - Heat and mass transport in nonhomogeneous random velocity fields. AB - The effective equation describing the transport of passive tracers in nonsolenoidal velocity fields is determined, assuming that the velocity field U(r,t) is a function of both position r and time t, albeit remaining locally random. Assuming a strong separation of scales and applying the method of homogenization, we find a Fickian constitutive relation for the coarse-grained particle flux, as the sum of a convective part, V(E)c, and a diffusive term, D(s). Inverted Delta c, where V(E) is the Eulerian mean tracer velocity, c the average particle concentration, and D(s) the effective diffusivity. The latter can be written as D(s)(r,t)=D(0)I+D(r,r,t), where D0 is the molecular diffusivity, I the unit dyadic and D(r(1),r(2),t) the cross diffusion dyadic. Conversely, the Eulerian mean velocity V(E)(r,t) is the sum of the microscale mean tracer velocity V(r,t) and a particle drift velocity, V(d)(r,t)= [(delta/delta r(2)).D(T)(r,r(2),t)](r(2)=r), which depends on the nonhomogeneity of the velocity field at the macroscale. The microscale mean particle velocity, in turn, is the sum of the mean fluid velocity and the ballistic tracer velocity, which is due to the local nonuniformity of the concentration field and is therefore structurally different from the tracer drift velocity. In the limit of large Peclet numbers, D(s) coincides with the self-diffusion dyadic, as it measures the local temporal growth of the mean square displacement of a tracer particle from its average position. In this case, the motion of a tracer particle is a random process in the manner of Stratonovich, where the smoothly varying mean tracer velocity equals the microscale mean tracer velocity and the fluctuating term is described through the cross diffusion dyadic D(r(1),r(2),t). PMID- 14754315 TI - Influence of electromagnetic boundary conditions onto the onset of dynamo action in laboratory experiments. AB - We study the onset of dynamo action of the Riga and Karlsruhe experiments with the addition of an external wall, the electromagnetic properties of which are different from those of the fluid in motion. We consider a wall of different thickness, conductivity, and permeability. We also consider the case of a ferrofluid in motion. PMID- 14754316 TI - Complete band gaps for liquid surface waves propagating over a periodically drilled bottom. AB - A plane-wave expansion approach is developed to solve the mild-slope equation for liquid surface waves propagating over a bottom with periodic structures. Band structures are calculated for the bottom periodically drilled with the square or triangular lattice of holes. Complete band gaps are found for both lattices. Parameters that influence the formation of band gaps are discussed. PMID- 14754317 TI - Maximum entropy theory and the rapid relaxation of three-dimensional quasi geostrophic turbulence. AB - Turbulent flow in a rapidly rotating stably stratified fluid (quasi-geostrophic turbulence) commonly decays toward a stable pattern of large-scale jets or vortices. A formula for the most probable three-dimensional end state, the maximum entropy state (MES), is derived using a form of Lynden-Bell statistical mechanics. The MES is determined by a set of integral invariants, including energy, as opposed to a complete description of the initial condition. A computed MES qualitatively resembles the quasistationary end state of a numerical simulation that is initialized with red noise, and relaxes for a time on the order of 100 (initial) eddy turnovers. However, the potential enstrophy of the end state, obtained from a coarsened potential vorticity distribution, exceeds that of the MES by nearly a factor of 2. The estimated errors for both theory and simulation do not account for the discrepancy. This suggests that the MES, if ever realized, requires a much longer time scale to fully develop. PMID- 14754318 TI - Influence of a nonlinear reference temperature profile on oscillatory Benard Marangoni convection. AB - We analyze oscillatory instabilities in a fluid layer of infinite horizontal extent, heated from above or cooled from below, taking into account the nonlinearity of the reference temperature profile during the transient state of heat conduction. The linear stability analysis shows that a nonlinear reference temperature profile can have a strong effect on the system, either stabilizing or destabilizing, depending on the relative importance of buoyancy and surface tension forces. For the nonlinear analysis we use a Galerkin-Eckhaus method leading to a finite set of amplitude equations. In the two-dimensional (2D) case, we show the solution of these amplitude equations are standing waves. PMID- 14754319 TI - Multistability of oscillatory thermocapillary convection in a liquid bridge. AB - A parametric investigation of the onset of chaos in a liquid bridge was numerically carried out for a medium Prandtl number liquid, Pr = 4, and unit aspect ratio under zero-gravity conditions. Spatiotemporal patterns of thermocapillary flow were successively studied beginning from the onset of instability up to the appearance of the nonperiodic flow and further on. Well tested numerical code is used for solving the three-dimensional time-dependent Navier-Stokes equations in cylindrical coordinate system. Two-dimensional steady flow becomes oscillatory with azimuthal wave number m=2 as a result of Hopf bifurcation at Re(cr)(1)=630. A second independent solution with wave number m=3 was found to appear at Reynolds number Re(cr)(2) approximately 810. Two branches of three-dimensional periodic orbits, traveling waves with m=2 and m=3, coexist for Re>Re(cr)(2). Additional stable branches do not connect them. The different flow organizations reveal different behaviors in the supercritical area. The m=2 traveling wave always remains periodic, but the mode m=3 starts exhibiting chaotic features at Re approximately 4200. The onset of temporal nonperiodicity was shown to be associated with development of broadband noise in spectra and preceded by a quasiperiodicity. The flow stabilizes back to periodic with single frequency when Re exceeds a value Re approximately 5100. The window of periodicity exists up to at least Re=6000, the largest investigated value of the Reynolds number. PMID- 14754320 TI - Destruction of stable spiral waves in oscillatory media. AB - We studied spiral wave dynamics in an oscillatory reaction diffusion system. We find a new phenomenon: without the appearance of any global modulation mode, stable spiral waves break up directly. By investigating the one-dimensional version of the system and the isolated local dynamics, we find that the unstable focus in the local dynamics plays an important role. For different boundary conditions (BCs), we find a transition between spiral waves and traveling waves for periodic BCs and drifting spiral wave for no-flux BC. PMID- 14754321 TI - Radiative opacities and configuration interaction effects of hot iron plasma using a detailed term accounting model. AB - We have calculated the radiative opacities of iron plasma in local thermodynamic equilibrium using a detailed term accounting model. The extensive atomic data are obtained by multiconfiguration Hartree-Fock (MCHF) method, with Breit-Pauli relativistic corrections. Extensive configuration interaction (CI) has been included based on LS coupling to obtain energy levels and the bound-bound transition cross sections. A detailed configuration accounting model is applied to evaluate the bound-free absorption cross sections. We simulate two experimental transmission spectra [G. Winhart et al., Phys. Rev. E 53, R1332 (1996); P. T. Springer et al., J. Quant. Spectrosc. Radiat. Transf. 58, 927 (1997)] to verify our calculation model, one is at a temperature of 22 eV and a density of 10(-2) g/cm(3) and the other is at a temperature of 20 eV and a lower density of 10(-4) g/cm(3). It is shown that the strong CI can effectively change the oscillator strengths in contrast to the single configuration HF method. For both of the two simulated transmission spectra good agreement is obtained between the present MCHF results and the experimental data. Spectrally resolved opacities and Planck and Rosseland mean opacities are also calculated. For the isothermal sequence of T=20 eV, when the density decreases from 10(-2) to 10(-5) g/cm(3), the linewidth also decreases so that the iron transition arrays show more discrete line structures and the linewidth becomes very important to the Rosseland mean opacity. PMID- 14754322 TI - Coupled dust-lattice solitons in monolayer plasma crystals. AB - Nonlinearly coupled dust-lattice (DL) waves in monolayer plasma crystals are studied theoretically. It is shown that the high-frequency transverse (vertical) oscillations can form localized wave envelopes--solitons coupled with "slow" longitudinal DL perturbations. Using the molecular dynamics simulations, the derived soliton solution is shown to be stable. PMID- 14754323 TI - Paraxial theory of slow self-focusing. AB - We present a theory of slow self-focusing that is paraxial in nature, gives the field including the phase and eikonal explicitly, while it also agrees with the results of variational and moments theories. After presenting the features of the theory, particularly its similarity to the central force problem, we go on to reformulate the theory for an absorbing medium. We find that the laser beam focuses to a constant beamwidth with a small phase-front curvature depending on the extent of absorption. The theory is applicable to a whole range of saturating nonlinearities although it specializes to two plasma cases, the ponderomotive force based and the relativistic electron quiver based nonlinearities, for definitive results. PMID- 14754324 TI - Longitudinal oscillations and linear Landau damping in quark-gluon plasma. AB - On the basis of the semiclassical kinetic Vlasov equation for quark-gluon plasma and the Yang-Mills equation in covariant gauge, linear Landau damping for electrostatic perturbations such as Langmuir waves is investigated for the extreme-relativistic and strongly relativistic cases. It has been observed that for the extreme-relativistic case, wherein the thermal speed of the particles exceeds the phase velocity of the perturbations, the linear Landau damping is absent as has been reported in the literature. However, a departure from extreme relativistic case generates an imaginary component of the frequency giving rise to linear Landau damping effect. The relevant integral for the conductivity tensor has been evaluated and the dispersion relation for the longitudinal part of the oscillation was obtained. Further, it is also noted that both the real part of the oscillation frequency and the damping rate are sensitive to the choice of the wave number k and the Debye length lambda(D) associated with quark gluon plasma. PMID- 14754325 TI - Normal mode spectra of two-dimensional classical atoms confined by a Coulomb potential. AB - The normal mode spectra of two-dimensional finite clusters of charged particles ( e) confined by a Coulomb potential resulting from a displaced positive charge Ze are obtained. This is a classical two-dimensional model system for atoms. We obtain the frequencies of the normal modes as a function of the confinement charge Z and the number of particles N. The analysis of the lowest normal mode frequency reveals a good agreement with the experimental results obtained in a system with screened interaction between charged particles. The dependence of the normal mode spectra as a function of a perpendicular magnetic field is also discussed and we found that the shear-like character of the modes is enhanced in the presence of the magnetic field. For large values of the magnetic fields the normal modes fall into two bands, a low frequency band with frequency approximately 1/omega(c) and a high frequency band with frequency approximately omega(c). PMID- 14754326 TI - Propagation of a short proton beam through a thin plasma slab. AB - A one-dimensional open boundary Vlasov code is used in order to investigate the propagation of a short proton beam through a plasma slab. Collisionless regimes are assumed, where the interaction between the beam and the plasma occurs due to the self-consistent, collective, electric field. Both charge compensated (by an accompanying electron cloud) and noncompensated beams are considered. PMID- 14754327 TI - Plasma state transition originating from local production of massive negative ions. AB - A dynamic evolution of plasma structures and its associated state transition due to local production of massive negative ions are investigated by the particle-in cell simulation which is based on an experimental configuration of an open plasma system (Q machine). Two different states, a stationary state and a dynamic state, are achieved depending on the production rate of massive negative ions. The plasma is stationary for a low production rate, while solitary pulses and fluctuations are spontaneously excited and the plasma state is unstable for a high production rate. Phenomena of a two-stream instability and a double layer formation induced by the local production are closely concerned with the excitation of the solitary pulse. The solitary pulses play a role in the rapid ejection of massive negative ions from the system. A staying time distribution of charged particles is analogically introduced for the special convenience of explanation, which corresponds with a so-called population pyramid. The staying time distribution can clearly express the feature of the state since all the phenomena occurring in the system directly reflect the distribution. PMID- 14754328 TI - Microscopic and macroscopic simulations for femtosecond-laser-matter interaction by cubic interpolated propagation method. AB - We performed 1.5-dimensional simulation for the Fokker-Planck equation using the CIP (cubic-interpolated propagation/constrained interpolation profile) method to investigate femtosecond-laser heating and transport processes. We found that the heat flux in the solid part approaches the Spitzer-Harm theory on quite a short time scale omega(p)t<50, and thus the subsequent evaporation process can be analyzed by classical thermal conduction. On the basis of this result, we performed a hydrodynamic simulation using the CIP method with classical thermal conduction in order to investigate the long time behavior of the evaporation process. The experimental ablation depth was replicated very well, showing that even femtosecond pulse laser processing can be satisfactorily described by classical heat conduction. Since the damage size is shown to be much larger than laser-spot size even in such an ultrashort-pulse experiment, we must use the fluence estimated by the damage size, which is twice as large as the laser spot, to correctly replicate the experiments. PMID- 14754329 TI - Fine structure of the Vavilov-Cherenkov radiation. AB - The aim of this paper is to study the fine structure of the Cherenkov rings. We analyze the experiments performed by one of authors (Zrelov) in which no special focusing devices were used. The broad Cherenkov ring was observed in the plane perpendicular to the motion axis. Using the exact and approximate formulas we investigate how a charge moving uniformly in a medium radiates in a finite space interval (the Tamm problem). The formulas obtained describe the radiation intensity in the whole space, inside and outside the Cherenkov ring. In the plane perpendicular to the motion axis, the radiation fills mainly the finite ring. Its width (proportional to the motion interval) and the energy released in this ring do not depend on the position of the observation plane. Outside the Cherenkov ring, the radiation intensity suddenly drops. Inside it, the radiation intensity exhibits small oscillations which are due to the interference of the Vavilov Cherenkov radiation and bremsstrahlung. The bursts of the radiation intensity at the ends of the Cherenkov ring are associated with the shock waves arising at the instant when the charge velocity coincides with the light velocity in a medium. For the chosen motion interval, the well-known Tamm formula does not describe the radiation intensity inside the Cherenkov ring for any position of the observation plane. Outside the Cherenkov ring, the Tamm formula is valid only at very large distances. Theoretical calculations are in satisfactory agreement with experimental data. Thus, the combined experimental and theoretical study of the unfocused Cherenkov rings allows one to obtain information on the physical processes accompanying the Cherenkov radiation in the finite spatial interval (bremsstrahlung, transition of the light velocity barrier, etc.). PMID- 14754330 TI - Optical bistability and tristability in nonlinear metal/dielectric composite media of nonspherical particles. AB - Based on a spectral representation method and a self-consistent mean field theory, we present a general framework to investigate the optical bistability in a nonlinear two-phase composite, where spheroidal metallic inclusions are randomly oriented and embedded in a dielectric host. The relation between the spatial average of the local field squared <[absolute value of E](2)>(i) (i=1,2) and the external field squared E20 is obtainable through the spectral density function which is predicted from our recently derived Maxwell-Garnett approximation. In addition to single optical bistability (OB), the appearance of double OB and optical tristability (OT) is reported, and the corresponding phase diagram is given. We find that the regions of the single OB, the double OB, and the OT are dependent on the shape and volume fraction of the metallic particles. Our method allows us to take one step forward to study some field-dependent effective optical properties, such as the refractive index, extinction coefficient as well as reflectance. The general framework is also applied to investigate exactly the solvable composites consisting of nonlinear spheroidal inclusions and linear dielectric host in the dilute limit. To this end, the present method is shown to be in excellent agreement with the exact solution. In addition, the present method predicts a larger threshold intensity than the variational approach. PMID- 14754331 TI - Wave-function reconstruction of complex fields obeying nonlinear parabolic equations. AB - We present a generalized Gerchberg-Saxton (GS) algorithm for reconstructing a (2+1)-dimensional complex scalar wave field which obeys a known nonlinear nondissipative parabolic differential equation, given knowledge of the wave-field modulus at three or more values of an evolution parameter such as time. This algorithm is used to recover the complex wave function of a (2+1)-dimensional Bose-Einstein condensate (BEC) from simulated modulus data. The Gross-Pitaveskii equation is used to model the dynamics of the BEC, with the modulus information being provided by a temporal sequence of simulated absorption images of the condensate. The efficacy of the generalized GS algorithm is examined for a wide range of simulation conditions, including strong nonlinearities, vortex states and Poisson noise. The general form of this algorithm, which allows one to reconstruct a time-dependent wave function, will be useful for studying the phase dynamics of topological defects in coherent quantum systems. PMID- 14754332 TI - Femtosecond pulse compression in pressure-gas cells filled with argon. AB - The nonlinear propagation of femtosecond pulses in pressure-gas cells filled with argon is investigated. By increasing the pressure for reaching peak power levels close to the threshold for self-focusing, it is shown that either group-velocity dispersion or multiphoton ionizing (MPI) sources can become key players for arresting the beam collapse. For input powers noticeably above critical, MPI rapidly dominates and the formation of self-guided filaments of light occurs. We discuss the dynamical role of MPI in shortening the pulse duration up to the optical cycle limit. Two different wavelength domains are commented. The influence of space-time focusing and self-steepening effects is furthermore discussed. Their respective roles in promoting shock structures are studied and shown to still promote pulse shortening in suitable power regimes. Finally, spectral broadening is analyzed and proven to be more important for large laser wavelengths. Numerical integration of the propagation equations is explained in the light of analytical arguments. PMID- 14754333 TI - Pulse delay and propagation through subwavelength metallic slits. AB - The transmission properties of a 2D metallic grating are investigated at optical wavelengths for an incident Gaussian pulse having pulse widths from 100 fs to 10 ps. The slits in the grating are subwavelength which can nevertheless allow significant transmission in the narrow wavelength regions where the so-called surface plasmon polariton (SPP) and waveguide mode resonances occur. The solution is obtained for each spectral component of the pulse by using the rigorous coupled wave approach and then the temporally varying output pulse is reconstructed by the standard method of taking an inverse Fourier transform. The delay of the pulse and the output pulse widths are determined by taking the first and second order moments of the Poynting vector with respect to time. It is shown that the time delay may be significant, as much as 256 fs for a pulse width of 200 fs for the SPP resonance but quite small (32 fs) for the waveguide mode resonance. The focus of the work is on demonstrating how the pulse delay evolves as the pulse propagates in the half-space beyond the grating. It is shown that the distance over which the time delay develops is much larger than the actual longitudinal dimension of the grating structure and it is approximately the same distance over which the stored energy and the vortices of the Poynting vector extend. PMID- 14754334 TI - Stable two-dimensional dispersion-managed soliton. AB - The existence of a dispersion-managed soliton in two-dimensional nonlinear Schrodinger equation with periodically varying dispersion has been explored. The averaged equations for the soliton width and chirp are obtained which successfully describe the long time evolution of the soliton. The slow dynamics of the soliton around the fixed points for the width and chirp are investigated and the corresponding frequencies are calculated. Analytical predictions are confirmed by direct partial differential equation (PDE) and ordinary differential equation (ODE) simulations. Application to a Bose-Einstein condensate in optical lattice is discussed. The existence of a dispersion-managed matter-wave soliton in such system is shown. PMID- 14754335 TI - Superluminal propagation of light pulses: A result of interference. AB - The propagation of pulses through dispersive media was investigated by solving Maxwell's equations without any approximation. We show that the superluminal propagation of pulses through anomalous dispersive media is a result of the interference of different frequency components composed of the pulse. The coherence of the pulse plays an important role for the superluminal propagation. With the decrease of the coherence of the pulse, the propagation changes from superluminal to subluminal. We have shown that the anomalous dispersion (the real part of the susceptibility) not the amplification (the imaginary part of the susceptibility) plays the essential role in the superluminal propagation. Although the superluminality always exists as long as the spectrum of the coherent pulse is within the anomalously dispersive region, both the energy propagation velocity and the frontal velocity never exceed the light speed in the vacuum. The output pulse through the medium is not the original pulse; instead it carries the information of the original pulse and the information of the prepared medium. PMID- 14754336 TI - Alternative coupled integrable optical soliton system with higher-order effects. AB - The system of coupled Hirota equations, which explains the simultaneous propagation of two fields in a nonlinear optical fiber with the inclusion of higher-order linear and self-steepening effects, is considered. By making use of a Painleve singularity structure analysis, the system is found to be an exactly integrable soliton system for three choices of physical parameters. Two of the soliton conditions are already well studied. For the third system, the soliton solutions are obtained using bilinear forms. PMID- 14754337 TI - Energy exchange in collisions of intrinsic localized modes. AB - Energy transfer process is examined numerically for the binary collision of intrinsic localized modes (ILMs) in the Fermi-Pasta-Ulam beta lattice. Unlike "solitons" in the integrable systems, ILMs exchange their energy in collision due to the discreteness effect. The mechanism of this energy exchange is examined in detail, and it is shown that the phase difference is the most dominant factor in the energy exchange process and, generally speaking, the ILM with advanced phase absorbs energy from the other. Heuristic model equations which describe the energy transfer of ILMs are proposed by considering the ILMs as interacting "particles." The results due to these equations agree qualitatively very well with those of the numerical simulations. In some cases, the relation between the phase difference of the ILMs and the transferred energy becomes singular, which may be regarded as one of the major mechanisms responsible for the generation of "chaotic breathers." PMID- 14754338 TI - Simulation study of localization of electromagnetic waves in two-dimensional random dipolar systems. AB - We study the propagation and scattering of electromagnetic waves by random arrays of dipolar cylinders in a uniform medium. A set of self-consistent equations, incorporating all orders of multiple scattering of the electromagnetic waves, is derived from first principles and then solved numerically for electromagnetic fields. For certain ranges of frequencies, spatially localized electromagnetic waves appear in such a simple but realistic disordered system. Dependence of localization on the frequency, radiation damping, and filling factor is shown. The spatial behavior of the total, coherent, and diffusive waves is explored in detail, and found to comply with a physical intuitive picture. A phase diagram characterizing localization is presented, in agreement with previous investigations on other systems. PMID- 14754339 TI - Perturbation-induced radiation by the Ablowitz-Ladik soliton. AB - An efficient formalism is elaborated to analytically describe dynamics of the Ablowitz-Ladik soliton in the presence of perturbations. This formalism is based on using the Riemann-Hilbert problem and provides the means of calculating evolution of the discrete soliton parameters, as well as shape distortion and perturbation-induced radiation effects. As an example, soliton characteristics are calculated for linear damping and quintic perturbations. PMID- 14754340 TI - Dynamic counterpropagating vector solitons in saturable self-focusing media. AB - We display rich spatial and temporal dynamics of light fields counterpropagating in a saturable self-focusing medium numerically, and analyze instabilities that counterpropagating solitons experience. An expression for the maximum length that the medium must not exceed for the solitons to be stable is derived and connected to the coupling strength of beam interaction. The instability can lead to periodic or irregular temporal dynamics of the light beams. By considering mutually incoherent counterpropagating beams, we show that differences to the copropagating case are due to the different boundary conditions. PMID- 14754341 TI - Spatial-temporal Gauss-Laguerre waves in dispersive media. AB - A family of dispersionless and diffractionless spatial-temporal Gauss-Laguerre waves propagating in dispersive linear and transparent media is introduced. Contrary to pulsed Bessel beams and envelope-X waves recently studied in media with normal dispersion, these spatiotemporal Gauss-Laguerre beams may exist both in the normal and anomalous dispersion spectral regions of the medium. PMID- 14754342 TI - Experimental studies of the influence of distributed power losses on the transparency of two-dimensional surface photonic band-gap structures. AB - Two-dimensional (2D) surface photonic band-gap (SPBG) structures have been suggested to realize 2D distributed feedback. The 2D SPBG structures can be obtained by providing 2D periodic perturbations of the waveguide surface. Such a structure can be used in a wide variety of applications including microwave electronics and integrated optics. The theoretically predicted effect of the transparency of the 2D SPBG structure when distributed Ohmic losses inside the structure are relatively high in comparison with the wave coupling coefficient has been observed in a series of experiments. The results obtained are in good agreement with theoretical predictions. PMID- 14754343 TI - Multireflection boundary conditions for lattice Boltzmann models. AB - We present a general framework for several previously introduced boundary conditions for lattice Boltzmann models, such as the bounce-back rule and the linear and quadratic interpolations. The objectives are twofold: first to give theoretical tools to study the existing link-type boundary conditions and their corresponding accuracy; second to design boundary conditions for general flows which are third-order kinetic accurate. Using these new boundary conditions, Couette and Poiseuille flows are exact solutions of the lattice Boltzmann models for a Reynolds number Re=0 (Stokes limit) for arbitrary inclination with the lattice directions. Numerical comparisons are given for Stokes flows in periodic arrays of spheres and cylinders, linear periodic array of cylinders between moving plates, and for Navier-Stokes flows in periodic arrays of cylinders for Re<200. These results show a significant improvement of the overall accuracy when using the linear interpolations instead of the bounce-back reflection (up to an order of magnitude on the hydrodynamics fields). Further improvement is achieved with the new multireflection boundary conditions, reaching a level of accuracy close to the quasianalytical reference solutions, even for rather modest grid resolutions and few points in the narrowest channels. More important, the pressure and velocity fields in the vicinity of the obstacles are much smoother with multireflection than with the other boundary conditions. Finally the good stability of these schemes is highlighted by some simulations of moving obstacles: a cylinder between flat walls and a sphere in a cylinder. PMID- 14754345 TI - Compact all-pass filters in photonic crystals as the building block for high capacity optical delay lines. AB - Optical all-pass filters, which generate strong on-resonance optical delay while maintaining a unity transmission coefficient throughout the entire resonant bandwidth, are of great importance for constructing delay lines in optical buffer applications. We provide an analysis of optical delay lines based upon cascading multiple stages of all-pass filter structures. We show that the maximum capacity of such delay lines is determined primarily by the dimensions of each stage. Motivated by this analysis, we describe compact optical all-pass filters in two dimensional photonic crystals. The accidental degeneracy of the cavity modes introduces a strong group delay and dispersion while maintaining total transmission. PMID- 14754344 TI - Soliton propagation in an erbium-doped fiber with and without a continuous wave background. AB - Considering ultrashort pulse propagation in a nonlinear resonant fiber governed by Hirota-Maxwell Bloch equations, the soliton interaction in an erbium-doped fiber system associated with higher-order dispersion, self-steepening, and self induced transparency effects is studied for the case when the fiber is driven with and without a constant pumping source. Using auto-Backlund-transformation, one- and two-soliton solutions are generated. The significance of the results is discussed in detail. PMID- 14754346 TI - Second-harmonic generation in one-dimensional photonic edge waveguides. AB - Diffraction losses in one-dimensional photonic crystal (PC) waveguides are the primary limitation on second-harmonic (SH) conversion efficiency. By using a finite difference time domain (FDTD) code taking into account second-order nonlinear polarization, we investigated these losses numerically, particularly at the SH wavelength. We propose an efficient SH conversion scheme in Al(x)Ga(1 x)As/air-etched waveguides. An analytical model is used to extrapolate the conversion efficiency to a number of periods for which time consumption makes the FDTD codes unsuitable. PMID- 14754347 TI - Imaging in turbid media using modulation frequency scanning. AB - We propose an imaging scheme to detect an object inside a turbid medium using intensity modulated waves as a probe. We show how the new degree of freedom represented by the modulation frequency allows sufficient control to reconstruct the image from the scattered wave signal. PMID- 14754348 TI - Probability of anomalously large bit-error rate in long haul optical transmission. AB - We consider a linear model of optical transmission through a fiber with birefringent disorder in the presence of amplifier noise. Both disorder and noise are assumed to be weak, i.e., the average bit-error rate (BER) is small. The probability distribution function (PDF) of rare violent events leading to the values of BER much larger than its typical value is estimated. We show that the PDF has a long algebraic-like tail. PMID- 14754349 TI - Towards idempotent reduced density matrices via particle-hole duality: McWeeny's purification and beyond. AB - Generalizations of McWeeny's purification formula are developed within the formalism of the particle-hole duality from the theory of reduced density matrices. Each of the generalized purification formulas is expressed as a sum of the one-particle reduced density matrix (1-RDM) and a finite series in the product of the one-particle and the one-hole RDMs, a product which vanishes in the limit that the 1-RDM is idempotent. Two categories of purification formulas are explored: (i) formulas which treat the "occupied" and the "virtual" occupation numbers equivalently and (ii) formulas which treat these occupation numbers differently. The latter category includes and extends the purification formulas derived in the context of the 1,2-contracted Schrodinger equation [D. A. Mazziotti, J. Chem. Phys. 115, 8305 (2001)]. While the McWeeny purification minimizes the absolute error in the occupation numbers quadratically, the generalized purification formulas exhibit faster than quadratic convergence of the 1-RDM towards idempotency. Application of these purification formulas in existing algorithms for linear scaling will be explored and discussed including illustrative calculations on sodium wires of length 10, 20, 30, and 40 atoms. PMID- 14754350 TI - Vapor-liquid coexistence in many-body dissipative particle dynamics. AB - Many-body dissipative particle dynamics is constructed to exhibit vapor-liquid coexistence, with a sharp interface, and a vapor phase of vanishingly small density. The application to fluid mechanics problems involving free surfaces is illustrated by simulation of a pendant drop. The model is an unusual example of a soft-sphere liquid with a potential energy built out of local-density-dependent one-particle self-energies. PMID- 14754351 TI - Fast and accurate coarsening simulation with an unconditionally stable time step. AB - We present Cahn-Hilliard and Allen-Cahn numerical integration algorithms that are unconditionally stable and so provide significantly faster accuracy-controlled simulation. Our stability analysis is based on Eyre's theorem and unconditional von Neumann stability analysis, both of which we present. Numerical tests confirm the accuracy of the von Neumann approach, which is straightforward and should be widely applicable in phase-field modeling. For the Cahn-Hilliard case, we show that accuracy can be controlled with an unbounded time step Delta t that grows with time t as Delta t approximately t(alpha). We develop a classification scheme for the step exponent alpha and demonstrate that a class of simple linear algorithms gives alpha=1/3. For this class the speedup relative to a fixed time step grows with N, the linear size of the system, as N/ln N. With conservative choices for the parameters controlling accuracy and finite-size effects we find that an 8192(2) lattice can be integrated 300 times faster than with the Euler method. PMID- 14754352 TI - Neuberger's double-pass algorithm. AB - We analyze Neuberger's double-pass algorithm for the matrix-vector multiplication R(H) x Y [where R(H) is (n-1,n)th degree rational polynomial of positive definite operator H], and show that the number of floating-point operations is independent of the degree n, provided that the number of sites is much larger than the number of iterations in the conjugate gradient. This implies that the matrix-vector product (H)(-1/2)Y approximately R((n-1,n))(H).Y can be approximated to very high precision with sufficiently large n, without noticeably extra costs. Further, we show that there exists a threshold n(T) such that the double-pass is faster than the single pass for n>n(T), where n(T) approximately 12-25 for most platforms. PMID- 14754353 TI - Interpreting the wide scattering of synchronized traffic data by time gap statistics. AB - Based on the statistical evaluation of experimental single-vehicle data, we propose a quantitative interpretation of the erratic scattering of flow-density data in synchronized traffic flows. A correlation analysis suggests that the dynamical flow-density data are well compatible with the so-called jam line characterizing fully developed traffic jams, if one takes into account the variation of their propagation speed due to the large variation of the netto time gaps (the inhomogeneity of traffic flow). The form of the time gap distribution depends not only on the density, but also on the measurement cross section: The most probable netto time gap in congested traffic flow upstream of a bottleneck is significantly increased compared to uncongested freeway sections. Moreover, we identify different power-law scaling laws for the relative variance of netto time gaps as a function of the sampling size. While the exponent is -1 in free traffic corresponding to statistically independent time gaps, the exponent is about -2/3 in congested traffic flow because of correlations between queued vehicles. PMID- 14754354 TI - Universality of anomalous one-dimensional heat conductivity. AB - In one and two dimensions, transport coefficients may diverge in the thermodynamic limit due to long-time correlation of the corresponding currents. The effective asymptotic behavior is addressed with reference to the problem of heat transport in one-dimensional crystals, modeled by chains of classical nonlinear oscillators. Extensive accurate equilibrium and nonequilibrium numerical simulations confirm that the finite-size thermal conductivity diverges with system size L as kappa proportional to L alpha. However, the exponent alpha deviates systematically from the theoretical prediction alpha=1/3 proposed in a recent paper [O. Narayan and S. Ramaswamy, Phys. Rev. Lett. 89, 200601 (2002)]. PMID- 14754355 TI - Low-temperature phase transition in the three-state Potts glass. AB - The low-temperature instability of one-step replica symmetry breaking (1RSB) phase in three-state Potts spin glass is obtained explicitly. The temperature of the instability is higher than the temperature where the 1RSB entropy becomes negative. The conjecture of the possibility of the low-temperature full RSB is supported. PMID- 14754356 TI - Current reversal with type-I intermittency in deterministic inertia ratchets. AB - The intermittency is investigated when the current reversal occurs in a deterministic inertia ratchet system. To determine which type the intermittency belongs to, we obtain the return map of velocities of particle by using stroboscopic recordings, and by numerically calculating the distribution of the average laminar length . The distribution follows the scaling law of proportional to epsilon(-1/2), the characteristic relation of type-I intermittency. PMID- 14754357 TI - Amplitude death induced by dynamic coupling. AB - The present paper shows that dynamic coupling induces amplitude death in coupled identical oscillators. For a simple limit-cycle oscillator, our theoretical analysis provides the necessary and sufficient condition for amplitude death. Furthermore, we guarantee that amplitude death never occurs, if each oscillator satisfies the odd number property that is known in the field of delayed-feedback control of chaos. PMID- 14754358 TI - Synchronization of fractional order chaotic systems. AB - The chaotic dynamics of fractional order systems began to attract much attention in recent years. In this Brief Report, we study the master-slave synchronization of fractional order chaotic systems. It is shown that fractional order chaotic systems can also be synchronized. PMID- 14754359 TI - Validity of numerical trajectories in the synchronization transition of complex systems. AB - We investigate the relationship between the loss of synchronization and the onset of shadowing breakdown via unstable dimension variability in complex systems. In the neighborhood of the critical transition to strongly nonhyperbolic behavior, the system undergoes on-off intermittency with respect to the synchronization state. There are potentially severe consequences of these facts on the validity of the computer-generated trajectories obtained from dynamical systems whose synchronization manifolds share the same nonhyperbolic properties. PMID- 14754360 TI - Non-normal and stochastic amplification of magnetic energy in the turbulent dynamo: subcritical case. AB - Our attention focuses on the stochastic dynamo equation with non-normal operator that gives an insight into the role of stochastics and non-normality in magnetic field generation. The main point of this Brief Report is a discussion of the generation of a large-scale magnetic field that cannot be explained by traditional linear eigenvalue analysis. The main result is a discovery of nonlinear deterministic instability and growth of finite magnetic field fluctuations in alpha beta dynamo theory. We present a simple stochastic model for the thin-disk axisymmetric alpha Omega dynamo involving three factors: (a) non-normality generated by differential rotation, (b) nonlinearity reflecting how the magnetic field affects the turbulent dynamo coefficients, and (c) stochastic perturbations. We show that even for the subcritical case (all eigenvalues are negative), there are three possible mechanisms for the generation of magnetic field. The first mechanism is a deterministic one that describes an interplay between transient growth and nonlinear saturation of the turbulent alpha effect and diffusivity. It turns out that the trivial state is nonlinearly unstable to small but finite initial perturbations. The second and third are stochastic mechanisms that account for the interaction of non-normal effect generated by differential rotation with random additive and multiplicative fluctuations. PMID- 14754361 TI - Nonlinearly coupled upper-hybrid and magnetoacoustic waves in collisional magnetoplasmas. AB - The nonlinear interaction between large amplitude electrostatic upper-hybrid (UH) waves and magnetoacoustic perturbations in a collisional magnetoplasma is considered. For this purpose, we present a nonlinear Schrodinger equation for the UH wave envelope in the presence of the electron density, electron temperature, and compressional magnetic field perturbations of the dispersive magnetoacoustic waves, and derive an equation for the latter taking into account the combined influence of the UH ponderomotive force and the differential electron Joule heating in the UH wave electric field. The coupled equations are employed to investigate modulational instabilities of a constant amplitude UH wave, and the density profile modifications due to the UH ponderomotive and thermal forces. It is found that the thermal instability has its maximum at a wavelength half the one of the UH wave, and a numerical solution of the coupled time-dependent system of equations reveals that localized density cavities are also created with a spacing approximately half the wavelength of the pump wave. PMID- 14754362 TI - Dust size distribution for dust acoustic waves in a magnetized dusty plasma. AB - A reasonable normalization for a magnetized dusty plasma with many different dust grains is adopted, which varies self-consistently with the system parameters. A Zakharov-Kuznetsov equation for small but finite amplitude dust acoustic waves is obtained for magnetized dusty plasma which contains different dust grains by using the reductive perturbation technique. We study the dust size distribution. Some comparisons are made between dusty plasma in which the dust size distribution is considered, and the monosized dusty plasma in which there is only one kind of dust grain whose size is the average dust size. This suggests that both soliton velocity and width are larger than that for monosized dusty plasma, but its amplitude is smaller than that for monosized dusty plasma. If there are positively charged dust grains, compressive solitary waves may exist. The velocity, amplitude, and width of a soliton in multidimensional form for a magnetized dusty plasma which contains many different dust grains are studied as well. PMID- 14754363 TI - Ablation pressure scaling at short laser wavelength. AB - The ablation pressure at a 0.44-microm laser wavelength has been measured at irradiance up to 2 x 10(14) W/cm(2). The diagnostics consisted in the detection of shock breakout from stepped Al targets. By adopting large focal spots and smoothed laser beams, the lateral energy transport and "drilling effects" have been avoided. The measured scaling shows a fair agreement with analytical models. PMID- 14754364 TI - Subwavelength imaging by a left-handed material superlens. AB - In this work, a finite-difference time-domain (FDTD) method is employed to justify the superlensing effect of left-handed material (LHM) slabs. Our results demonstrate that subwavelength resolution can be achieved by LHM slabs with certain parameters. We present the dynamic feature of the imaging process and the dependence of physical parameters on the performance of the superlens. These results help to clarify the diverse FDTD results reported previously. We also show that the achievable resolution is limited by the absorption and thickness of the LHM slabs, which introduces difficulties in practical applications of the superlens. PMID- 14754365 TI - Comment on "Hysteresis phenomena in CO catalytic oxidation system in the presence of inhomogeneities of the catalyst surface". AB - Scrutinizing the Monte Carlo algorithm, used by D.-Y. Hua and Y.-Q. Ma [Phys. Rev. E 66, 066103 (2002)] in order to simulate the effect of defect sites on bistable kinetics of CO oxidation on single-crystal surfaces, we show that in their study (i) the rules for describing CO adsorption, desorption, and surface diffusion contradict the detailed balance principle and (ii) the ratio of the rates of CO diffusion and reaction between adsorbed CO and O species is opposite compared to that observed in reality. PMID- 14754373 TI - Opioid receptors and their ligands. AB - This review gives a historical perspective, summarizing approximately 25 years of research on opioids. The "typical" opioid peptides produced in the brain, "atypical" opioids encrypted in milk protein or hemoglobin sequences, and extremely potent and selective opioids of amphibian origin are described. The main focus is on the structure-activity relationship studies of peptide ligands for three main opioid receptor types (micro, delta, kappa), their selectivities and pharmacological activities in vitro. Chemical modifications that led to obtaining potent and selective agonists and antagonists for these receptors are discussed. PMID- 14754374 TI - New trends in the development of opioid peptide analogues as advanced remedies for pain relief. AB - The search for new peptides to be used as analgesics in place of morphine has been mainly directed to develop peptide analogues or peptidomimetics having higher biological stability and receptor selectivity. Indeed, most of the alkaloid opioid counterindications are due to the scarce stability and the contemporary activation of different receptor types. However, the development of several extremely stable and selective peptide ligands for the different opioid receptors, and the recent discovery of the micro-receptor selective endomorphins, rendered this search less fundamental. In recent years, other opioid peptide properties have been investigated in the search for new pharmacological tools. The utility of a drug depends on its ability to reach appropriate receptors at the target tissue and to remain metabolically stable in order to produce the desired effect. This review deals with the recent investigations on peptide bioavailability, in particular barrier penetration and resistance against enzymatic degradation; with the development of peptides having activity at different receptors; with chimeric peptides, with propeptides, and with non conventional peptides, lacking basic pharmacophoric features. PMID- 14754375 TI - Endogenous opioids and addiction to alcohol and other drugs of abuse. AB - There is significant experimental evidence implicating the endogenous opioid system (opioid peptides and opioid receptors) with the processes of reward and reinforcement. Indeed, many behaviors associated with reward and reinforcement, for example feeding behavior, are controlled by distinct components of the endogenous opioid system located in relevant brain regions. It has also been shown that regardless of their initial site of action many drugs of abuse, such as morphine, nicotine, cocaine, alcohol and amphetamines, induce an increase in the extracellular concentration of dopamine in the nucleus accumbens. This increased secretion of dopamine in the nucleus accumbens seems to be a common effect of many drugs of abuse, and it was proposed that may mediate their rewarding and reinforcing properties. Furthermore, activation of micro opioid receptors in the ventral tegmental area, or of micro and delta opioid receptors in the nucleus accumbens enhances the extracellular concentration of dopamine in the nucleus accumbens. Thus, stimulation of the activity of distinct components of the endogenous opioid system either by opioid or by other drugs of abuse, may mediate some of their reinforcing effects. In this review article, a brief description of the endogenous opioid system and its implication in the processes of reward and reinforcement of opioid and other drugs of abuse will be presented. Furthermore, the use of opioid antagonists in the treatment of drug addiction will be discussed. Special emphasis will be given to ethanol addiction, the drug mainly studied in my laboratory. PMID- 14754376 TI - Expression of opioid receptors during peripheral inflammation. AB - Opioid receptors (OR) and their mRNA are present in the central and peripheral nervous system of mammals. In this review we examine the behavioral effects of opioids and the expression of their receptors during peripheral inflammation in two experimental models: the rat paw and the mouse intestine. Inflammation increased the antinociceptive (paw) and the inhibitory effects of opioids in the gut (transit, permeability and plasma extravasation) by interaction with OR located at peripheral sites. Based on agonist efficacy, micro > delta >> kappa-OR mediate the antinociceptive and antitransit effects of opioids during inflammation. Intestinal permeability is modulated by delta = micro >> kappa-OR, while kappa > delta >> micro-OR are involved in the inhibition of plasma extravasation. Intestinal inflammation increased the transcription of micro and delta-OR (but not kappa) genes in the gut, thus explaining the enhanced antitransit and antisecretory effects of micro and delta-OR agonists; however, the increased inhibitory effects of kappa-OR agonists on plasma extravasation could result from post-transcriptional regulation of the receptor. Similarly, the increased expression of peripheral micro-OR observed in the rat paw during inflammation, occurs at post-transcriptional levels and is related to an increased axonal transport from the dorsal root ganglia to peripheral terminals. The sites and mechanisms implicated in the increased transcription of micro and delta-OR during intestinal inflammation are under investigation. PMID- 14754377 TI - Neuropeptides and gastric mucosal homeostasis. AB - The role of central nervous system (CNS) in regulation of gastric function has long been known. The dorsal vagal complex (DVC) has an important role in regulation of gastric mucosal integrity; it is involved both in mucosal protection and in ulcer formation. Neuropeptides have been identified in DVC, the origin of these peptides are both intrinsic and extrinsic. Neuropeptides are localized also in the periphery, in afferent neurons. The afferent neurons also have efferent-like function in the gastroinetestinal tract, and neuropeptides released from the peripheral nerve endings of primary afferent neurons can induce gastric mucosal protection. Centrally and /or peripherally injected neuropeptides, such as amylin, adrenomedullin, bombesin, cholecystokinin, neurotensin, opioid peptides, thyreotropin releasing hormone and vasoactive intestinal peptide, influence both the acid secretion and the gastric mucosal lesions induced by different ulcerogens. The centrally induced gastroprotective effect of neuropeptides may be partly due to a vagal dependent increase of gastric mucosal resistance to injury; activation of vagal cholinergic pathway is resulted in stimulation of the release of mucosal prostaglandin and nitric oxide. Furthermore, release of sensory neuropeptides (calcitonin gene-related peptide, tachykinins) from capsaicin sensitive afferent fibers are also involved in the centrally induced gastroprotective effect of neuropeptides. PMID- 14754378 TI - Substance P: structure, function, and therapeutics. AB - Extensive efforts since 1931, on the structural determination of the mammalian tachykinin SP by NMR, CD and IR have turned out to be inconclusive. Studies are now being concentrated on the structural properties and characteristics of various NK receptors (NK(1), NK(2) and NK(3)) with the help of genetics, cloning, receptor engineering, mutagenesis and modeling. This knowledge is now being fruitfully used in the development of non-peptide NK(1) receptor antagonists that essentially block the pharmacological effects of SP. It is now being realized that the simultaneous blockade of two or more receptors gives promising results in emesis, depression and pulmonary obstructive diseases. In addition to the synthetic compounds, the discovery of antagonists from natural origin has added a great value to this field. In this review we have made an attempt to present the structural characteristics of SP, its analogs and antagonists, the structural characteristics of the NK receptor, and structure activity relationships that have helped to improve the therapeutic utilities of SP antagonists. PMID- 14754379 TI - Recent advances in the investigation of the bioactive conformation of peptides active at the micro-opioid receptor. conformational analysis of endomorphins. AB - Despite of the recent advances in the structural investigation of complex molecules, the comprehension of the 3D features responsible for the interaction between opioid peptides and micro-opioid receptors still remains an elusive task. This has to be attributed to the intrinsic nature of opioid peptides, which can assume a number of different conformations of similar energy, and to the flexibility of the receptorial cavity, which can modify its inner shape to host different ligands. Due to this inherent mobility of the ligand-receptor system, massive efforts devoted to the definition of a rigid bioactive conformation to be used as a template for the design of new pharmacologically active compounds might be overstressed. The future goal might be the design of peptide or nonpeptide ligands capable of maximizing specific hydrophobic interactions. This review covers the recent opinions emerged on the nature of the ligand-receptor interaction, and the development of suitable models for the determination of the bioactive conformation of peptide ligands active towards micro-opioid receptors. PMID- 14754380 TI - Use of NMR and fluorescence spectroscopy as well as theoretical conformational analysis in conformation-activity studies of cyclic enkephalin analogues. AB - In this review the conformational studies of natural enkephalins (H-Tyr-Gly-Gly Phe-Met-OH; the [Met(5)]enkephalin and H-Tyr-Gly-Gly-Phe-Leu-OH; the [Leu(5)]enkephalin), their acyclic and cyclic analogues, including those carried out in our laboratory, performed by experimental and theoretical methods and their combination, are described. Emphasis is given on the role of conformational constraints introduced by cyclization on activity at the micro and delta opioid receptors. Comparison of the conformations of cyclic enkephalin analogues with high delta-receptor activity with those of potent rigid non-peptide delta receptor agonists indicates that the proximity of the aromatic side chains in positions 1 and 4 as well as the N-terminal amino group is desirable for the activity at the delta opioid receptors; early conformational studies also suggested that spatial separation of the aromatic side chains and rigidity of the cyclic backbone is desirable for micro-receptor activity. The results of our recent conformational studies performed with the use of fluorescence and NMR spectroscopy as well as theoretical calculations indicate, however, that these structural features are not necessary for activity at the micro opioid receptors. Methods applied to the determination of the conformation of flexible peptides, such as Nuclear Magnetic Resonance (NMR), fluorescence spectroscopy, and theoretical conformational analysis are also discussed briefly. PMID- 14754381 TI - Conformational analysis of opioid peptides in the solid states and the membrane environments by NMR spectroscopy. AB - Determination of conformations and structures of opioid peptides in the membrane environments is an essential step to understand the action of the peptide to the specialized receptors. This information not only gains insight into the structure function relationship of opioid peptide but also gives proper guidelines to design a new drug to have same neuroendocrine functions. This review provides the structural studies of three types of opioid peptide families such as enkephalin, beta-endorphin and dynorphin in the solid states and the membrane environments. The structures of enkephalins show that they take beta-bend, extended and double beta-bend structures in the crystals. Moreover, enkephalin molecules take a variety of structures in the crystals and are easily converted to the other structures with slightly different torsion angles. On the other hand, beta-bend structures are mostly seen in the membrane environments. Membrane bound structure of dynorphin shows that the N-terminus forms alpha-helical structure and is inserted into the membrane with the helical axis almost perpendicular to the membrane surface. It is discussed that the helical region of the extracellular loop II of the kappa-opioid receptor may interact with the helical region of dynorphin with a high affinity in the membrane environments. beta-endorphin takes alpha-helical structure at N-terminus and the central regions and the rest of regions take unordered structure when the bind to the membrane. Since the membrane bound structures of opioid peptides differ from those of the solution states, membrane association is an important process for exerting the affinity and the selectivity to the specific opioid receptors. PMID- 14754382 TI - Antagonism in opioid peptides: the role of conformation. AB - The availability of new, highly selective antagonists, in the field of opioid peptides and of other pain peptides, is important both for a better understanding of the interaction of the receptors with their ligands and for their practical relevance. The design of antagonists is not obvious even when the essential features of agonists are well known. In this review we have examined the main aspects of the problem using, as leading criteria two theoretical models of antagonism and the subdivision of opioid peptides into two functional domains. The main causes of antagonism have been integrated in two very general models: one, referred to as the participation model, attributes antagonism to the lack, with respect to the parent agonist, of an essential group, whereas another model, attributes antagonism to the misfit of the molecule inside the receptor. The second criterion is the division of the structure of peptide hormones, originally put forward by Robert Schwyzer, in two functional domains, the message domain, which is responsible of the larger part of the binding affinity of opioid agonists, and an address domain, which dictates most of the peptide specificity. The most significant achievements in the design of opioid antagonists are classified according to the relative importance of chemical constitution, conformation and chirality. PMID- 14754383 TI - Targets of antidementive therapy: drugs with a specific pharmacological mechanism of action. AB - Diagnosis and therapy of dementia have made considerable progress in recent years. Drugs have been developed which improve cognitive performance, delay the loss of abilities of daily living and prevent early nursing home placement in a considerable number of patients. With the various pharmacological and non pharmacological approaches, effective treatment options of AD are available at present and the therapeutic potential will even increase in future. Thus, the treatment of dementia should more focused and explicit in its goals for the doctor, the patient and the relatives. Therapeutic targets must be defined on the basis of the individual needs and deficits and with regard to different levels of the disease process. Ideally, treatment should always aim at an etiological and/or a pathophysiological level. At present, however, aiming at the neurotransmitter level, the core syndrome of cognitive deficits can be approached by treatment options. Further therapeutic targets can be defined on the level of activities of daily living, following a resource focused approach, as well as on the level of behavioral disturbances. Additional therapeutic targets should be seen under a humanitarian or palliative perspective. And finally, family members are also targets for therapy in dementia, even if such therapy is not directed towards the demented patient. All these treatment targets have to be evaluated and adapted under the perspective of time because prominent symptoms in AD change considerably with disease progression. Selection and adaptation of medication becomes easier if such targets are considered and if therapeutic effects are monitored target-specifically. PMID- 14754384 TI - Acetylcholinesterase inhibition in Alzheimer's Disease. AB - Alzheimer's Disease (AD) is the most common cause for dementia in our ageing population, which leads to a slowly progressive, irretrievable ruination of mental function. The destructive, primarily degenerative condition is neuropathologically characterized by the formation of amyloid plaques, neurofibrillary tangles and loss of neurons and synapses as well. Research during the past twenty years revealed early in the disease course a degeneration of cholinergic nuclei localised in the basal forebrain. Impairment of this cholinergic system, which projects into large areas of the limbic system and the neocortex is followed by disturbance of attentional processes and cognitive decline. The link between the cholinergic dysfunction and cognitive impairment has focused large scientific efforts to understand the neurobiology of cognition and to develop therapeutic tools for the fight against Alzheimer's Disease. Acetylcholinesterase inhibitors are currently the best established treatment for this devastating disease. This review describes historical aspects and the vast range of use of cholinesterase inhibitors in traditional societies and industrial nations. Second, the rational basis will be outlined for their development as medication, the so-called cholinergic hypotheses of AD. Third, acetylcholinesterase inhibitors currently available for the treatment of AD will be reviewed. This includes donepezil, galanthamine and rivastigmine. Tacrine, the first acetylcholinesterase inhibitor who became available in 1993 as a treatment for AD, does not play an essential role anymore besides his historical value, because of its hepatotoxicity. Although acetylcholinesterase inhibitors are no cure, these drugs can delay the progress of mental deterioration, reduce neuropsychiatric symptoms and therefore represent a rational therapeutic approach to the treatment of Alzheimer's Disease. PMID- 14754385 TI - NMDA-antagonism (memantine): an alternative pharmacological therapeutic principle in Alzheimer's and vascular dementia. AB - Memantine, a non-competitive NMDA antagonist, has been clinically used in the treatment of dementia in Germany for over ten years. The rationale for this indication is strongly related to the physiological and to the pathological role of glutamate in neurotransmission. Physiologically, NMDA receptors mediate synaptic plasticity by acting as a coincidence detector. Only those synapses that show temporally and spatially discrete activation of NMDA receptors undergo plastic changes secondary to Ca++ influx after rapid unblocking of Mg++, thus crucially contributing to memory and learning processes. The voltage-dependency of Mg++ is so pronounced that under pathological conditions it leaves the NMDA channel upon moderate depolarisation, thus interrupting memory and learning. Its pharmacological properties allow memantine to rapidly leave the NMDA channel upon transient physiological activation by synaptic glutamate (restoring significant signal transmission), but to block the sustained activation of low glutamate concentration under pathological conditions, i.e. to protect against excitotoxicity as a pathomechanism of neurodegenerative disorders. Memantine acts as a neuroprotective agent in various animal models based on both neurodegenerative and vascular processes as it ameliorates cognitive and memory deficits. Memantine has shown to be effective and safe in the treatment of dementia, particularly Alzheimer's disease, in controlled clinical trials. Provided that the dose is slowly increased it is generally well tolerated and safe up to 20 and 30 mg per day, with intake preferably in the morning. The compound is completely absorbed after oral intake with Cmax values after 6 hours, undergoes little metabolism and has a terminal elimination half life between 60 and 100 hours. Due to its low potential of interaction, memantine can be combined with acetylcholinesterase inhibitors, the mainstay of current symptomatic treatment of Alzheimer's disease and it is suited in elderly patients receiving multiple drug therapy. PMID- 14754386 TI - Review about Ginkgo biloba special extract EGb 761 (Ginkgo). AB - Ginkgo biloba extracts (EGb) are well-defined plant extracts. It has several indications as dementia, macula degeneration, tinnitus and winter depression. A review of the current and past literature about older people with Alzheimer's dementia or vascular dementia or age-associated memory impairment treated with Ginkgo biloba extract, reveals that EGb has reproducible effects on cognitive functions in Alzheimer's disease. The drug is well tolerated. PMID- 14754387 TI - Functional neurochemistry of Alzheimer's disease. AB - A review of neurochemical research on classical neurotransmitters, i.e. acetylcholine, serotonin, noradrenaline, dopamine, glutamate, and GABA in Alzheimer's disease is presented. Findings are linked to the information processing system of the human brain to establish a more functional neurochemistry. On this basis, different pharmacotherapeutic strategies are discussed. Our conclusion is that current symptomatic therapy of Alzheimer's disease is insufficient. Besides therapy with acetylcholinesterase inhibitors, comedication to act on imbalances between serotonin and noradrenaline on the one site, and dopamine, glutamate and GABA on the other site should should be considered. PMID- 14754388 TI - Economic aspects on drug therapy of dementia. AB - The great number of people suffering from dementia present a great challenge for the health care and social support systems. In a situation where resources are scarce, health economical aspects of dementia care are of great importance in order to identify cost-effective care. However, the literature in this field is limited. There are also aspects where there methodological development is necessary (e.g. informal care, quality of life, long term effects). In the absence of prospective long term data, it is necessary to use pharmacoeconomical models. It is also important to have population based data for the description of how resources and costs are allocated between the different care sectors. Current data show that there is a strong relationship between cognitive functioning and costs. The number of pharmacoeconomical evaluations of drugs influencing on the symptomatology of dementia is low. Available data show that there is support for a view that treatment is cost-neutral or perhaps cost saving which in combination with positive effects in terms of efficacy may indicate cost-effectiveness. PMID- 14754389 TI - Cholecystokinin antagonists a new way to improve the analgesia from old analgesics? AB - Cholecystokinin, originally thought to be confined only to the gastrointestinal tract, is now known to be co-localised in both the gastrointestinal tract and central nervous system. In animal models levels are increased after neural injury and with opioid administration. This peptide acts as an anti-opioid, and as levels increase, the extent of opioid derived antinociception decreases. Co administration of a CCK antagonist along with an opioid is associated with an improved level of antinociception. Furthermore CCK antagonists may prevent antinociceptive tolerance with opioids and even reverse established tolerance. Human studies have now confirmed the pro-analgesic effect of some CCK antagonists. Human investigation of the effect of CCK antagonists on analgesic tolerance has yet to be performed. This review examines the available evidence that suggests a role for CCK antagonists in human pain management. PMID- 14754390 TI - Clinical development of microbicides for the prevention of HIV infection. AB - The HIV/AIDS pandemic continues its spread at a rate of over 15,000 new infections every day. Sexual transmission of HIV-1 is the dominant mode of this pandemic spread. For the first time since the disease emerged in the early 1980s, about half the 42 million people now living with HIV/AIDS worldwide are women. Worldwide, more than 90 percent of all adolescent and adult HIV infections have resulted from heterosexual intercourse. The "feminization" of the pandemic largely driven by the social, economic, and biological factors warrants urgent attention particularly for the adolescent female population. In the absence of an effective prophylactic anti-HIV therapy or vaccine, current efforts are aimed at developing intravaginal/intrarectal topical formulations of anti-HIV agents or microbicides to curb the mucosal and perinatal HIV transmission. Microbicides would provide protection by directly inactivating HIV or preventing HIV from attaching, entering or replicating in susceptible target cells as well as dissemination from target cells present in semen or the host cells that line the vaginal/rectal wall. Thus, ideally, anti-HIV microbicides should be capable of attacking HIV from different angles. In addition, a contraceptive microbicide could help prevent unintended pregnancies worldwide. To be a microbicide, these agents must be safe, effective following vaginal or rectal administration, and should cause minimal or no genital symptoms following long-term repeated usage. A safe and efficacious anti-HIV microbicide is not yet available despite the fact that more than 60 candidate agents have been identified to have in vitro activity against HIV, 18 of which have advanced to clinical testing. Targeting HIV entry has been a favored approach because it is the first step in the process of infection and several readily available anionic polymeric products seem to variably interfere with these processes are the primary candidates for potential microbicides. Formulations of some anionic polymeric antiviral agents have been tested at various doses and various durations for safety, tolerability, and acceptability in Phase I/II clinical trials (vaginal, rectal, or penile studies) in HIV-uninfected and/or HIV-infected populations. Current multicenter Phase I/II safety and Phase II/III efficacy studies that are being conducted or planned in different geographical locations by various special interest groups are designed for rapid clinical development of candidate products. The currently marketed detergent-type spermicide, nonoxynol-9 (N-9), has failed in Phase III clinical trials, due to the drug-induced formation of localized genital lesions that might in fact actually promote virus transmission. Alternative "first-generation" microbicides that have undergone Phase I/II safety and tolerability studies in HIV-uninfected and/or HIV-infected volunteers include polymeric viral fusion inhibitors (dextrin sulfate/Emmelle, carrageenans [PC-213, PC-503, PC 515/Carraguard], cellulose sulfate/Ushercell, polystyrene sulfonate, naphthalene sulfonate [PIC 024-4/PRO 2000/5], acidifying gel [Carbomer 974P/BufferGel], Lactobacillus (L. crispatus) suppository/CTV-05, detergent-type dual-function barriers [ACIDFORM, GEDA Plus, SURETE, Glyminox/C31G/Savvy, Invisible Condom], herbal extracts [Praneem], and viral replication inhibitors [PMPA/Tenofovir]. For majority of these products, no information is available regarding their long-term mucosal safety, carcinogenicity potential, bioavailability, or efficacy following their extended vaginal or rectal exposure. The irritative genitourinary symptoms reported for a number of these first-generation products in Phase I clinical trials implies that the "soft" preclinical endpoints for mucosal safety established for the use and development of vaginal spermicides may not be rigorous enough for vaginal and rectal microbicides because of the efficient sexual tra virus diversity, and genetic environment. It is now apparent that sexually transmitted R5 HIV-1 viruses have less positive charge on their surface compared with the R4 HIV-1 viruses, which may limit the anionic polymers as topical microbicides despite extensive clinical trials. Nevertheless, their ongoing clinical trials, reviewed here, using optimized formulations, and special populations in various geographic locations are paving the way for future rigorous clinical testing of "mechanism-based" broad-spectrum anti-HIV microbicides that are currently under intense development. It is anticipated that future microbicide trials will focus on combination of products capable of attacking HIV life cycle at multiple steps intended to increase efficacy, limit cross-resistance as well as minimize microbicide-induced host toxicity. PMID- 14754392 TI - Control of autoimmune diseases by the B7-CD28 family molecules. AB - The roles of B7-CD28 family molecules in the regulation of immune responses have been intensively studied over the past decade. The findings resulting from these studies not only broaden our understanding in the control of immune responses at the molecular level, but also lead to identification of molecular targets for future manipulation and potential treatment of human diseases. There is convincing evidence that the B7-CD28 family molecules play critical roles in the control of initiation, progression and pathogenesis of autoimmune diseases, which is the focus of this review. Currently, five molecular pathways within this family have been identified and each of them appears to overlap but have distinct functions in the control of priming, activation, maturation and amplification of cellular and humoral immune responses. Rationale-based design of intervention, targeting on multiple pathways should lead to new methods and approaches for management of autoimmune diseases. PMID- 14754391 TI - Functional consequences of immune cell adhesion to endothelial cells. AB - Research regarding the interactions between the endothelium and immune cells has undergone a significant expansion during the past decade. Major shifts of emphasis have been the norm, from the production of a detail catalog of the cell surface receptors and counter-receptors acting at the interface between the vascular endothelium and circulating cells to a more mechanistic account of leukocyte/endothelium interactions. The past five years has seen new, groundbreaking developments in the field, with exiting studies aimed at understanding the functional consequences of the direct contact of endothelial cells and leukocytes. Based on early work to be discussed below, new data on local chemokine production and cell-to-cell contacts, attempt to clarify the physiopathological significance of these events. The exceptional anatomical arrangement of endothelial cells insures a permanent contact of the endothelium with leukocytes, an event likely to result in cellular signals originating from direct cell contact or through the action of soluble factors produced by endothelial cells or immune cells. As we will discuss, current evidence supports the idea that endothelial cells present at vascular endothelium as well as at specialized high endothelial venules, play not only a critical role in the homing and recruitment of immune cells but that it can also influence the outcome of the immune response. Additionally, new evidence clearly corroborates the idea that B and T lymphocytes as well as NK cells can modulate endothelial cell function. PMID- 14754393 TI - Endothelial expression of MHC class II molecules in autoimmune disease. AB - Major histocompatibility complex (MHC) class II molecules are up-regulated on endothelial cells in human allografts, and are thought to be involved in graft rejection. The MHC class II subtypes HLA-DR, DQ and DP regulate T cell dependent immune responses, and aberrant expression could be important in autoimmunity. Increased endothelial MHC class II expression has been demonstrated in several autoimmune diseases, including myocarditis with dilated cardiomyopathy, rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE). Recent data suggest that there is an association between endothelial expression of MHC class II molecules and diffuse endothelial dysfunction, which may be part of the explanation of the increased risk of cardiovascular disease in patients with RA, SLE and other chronic inflammatory conditions. MHC class II transcription is in part genetically determined. Cytokine induced up-regulation of MHC class II molecules can be inhibited in vitro by antioxidants and different drugs, such as cyclosporin and statins. Research on the development of new treatments for systemic autoimmune diseases and cardiovascular disease should include evaluation of effects on endothelial activation, including MHC class II expression. This review also discusses the genetic basis of MHC class II expression and its implications for understanding MHC genotype associations with autoimmune diseases. Recent studies of interactions between endothelial cells and T cells are reviewed. Such interactions could be of major importance in the pathogenesis of autoimmune and vascular diseases. PMID- 14754394 TI - Endothelial chemokines in autoimmune disease. AB - Compelling evidence now exists supporting the involvement of chemokines in the pathogenesis of autoimmune diseases. Examples of chemokines and chemokine receptors being involved in mediating autoimmune disease exist for rheumatoid arthritis, multiple sclerosis, allograft rejection, systemic lupus erythematosus, psoriasis, atopic dermatitis, lichen planus, and graft-versus-host-disease. Expression of chemokines by endothelial cells appears to be an important step in the development of these diseases. Since chemokines are small molecular weight molecules that act through G-protein coupled receptors, they make attractive drug targets. Several antagonists of chemokine - chemokine receptor interactions have been used to successfully alleviate some or all of the symptoms associated with many of these diseases in animal models. Further investigation of the involvement of chemokines in the pathogenesis or progression of autoimmune diseases may lead to practical clinical advances in diagnosis, prognosis, and therapy of such diseases. PMID- 14754395 TI - Regulation of B cell activation by PECAM-1: implications for the development of autoimmune disorders. AB - Regulation of B-cell development and activation is imperative to the myriad of activities that perpetuate humoral immunity. This T-cell dependent immune mechanism often relies upon the maintenance of T-cell tolerance, such that the maturity of the antigen-presentating cell, its function and molecular mimicry are contributing factors. Recent findings have implicated the involvement of the B cell and their corresponding surface co-receptors in regulating autoimmune disease. One candidate receptor, PECAM-1, has demonstrated the ability to downregulate both B and T-cell signalling pathways. The deletion of PECAM-1 in mice has led to a hyper-responsive B-cell phenotype with abnormal B-cell development. Additionally, in vivo functional studies have found that absence of PECAM-1 results in an increased susceptibility to autoimmune disorders of encephalomyelitis and Type I hypersensitivity reactions. Taken together, these findings indicate that PECAM-1 may have an important role in maintaining B-cell tolerance and regulatory function in preventing the onset of autoimmune disease. Elucidating the mechanisms of PECAM-1 function in autoimmune disorders could facilitate development of novel therapeutics. PMID- 14754396 TI - Dendritic cell endothelium interaction in autoimmunity. AB - Monocyte derived dendritic cells play a central role in controlling immunity by activating naive T lymphocytes. Monocytes can leave the blood stream by endothelial cell transmigration and differentiation into dendritic cells. A fundamental aspect of dendritic cell biology is their capacity to engulf tissue antigens and revers-migrate into lymph nodes. In lymph nodes dendritic cells can traffic to T-cell areas where they activate naive T-cells. Throughout this review we are developing a model of in vivo activation of auto-reactive T-cells by activated dendritic cells. PMID- 14754397 TI - Erythropoietin withdrawal leads to the destruction of young red cells at the endothelial-macrophage interface. AB - Erythropoietin is a growth factor for endothelial cells as well as for erythroid cells. In contrast to their proliferative response to physiological levels of erythropoietin, endothelial cells may respond to decreased levels by triggering a process called neocytolysis. Neocytolysis is the selective destruction of the youngest circulating red cells, which may be prompted by endothelial cells communicating with macrophages to stimulate phagocytosis of this unusual cell subset. We speculate that this is due to decreased production by endothelial cells of the macrophage-deactivating transforming growth factor-beta. The resulting proinflammatory phenotype may include macrophage production of thrombospondin, which forms bridges between adhesion molecules selectively expressed on young red cells (CD36) and the CD36/alphavbeta3 complex on macrophages that triggers phagocytosis. Alternatively, inflammatory mediators secreted by endothelial cells and macrophages during erythropoietin withdrawal may signal young red cells to expose phosphatidylserine, which would mark them for elimination via the normal pathway for aged red cell destruction. Neocytolysis has been demonstrated in returning astronauts and in polycythemic individuals at high altitude on descent to sea level. It contributes to the anemia of renal disease, is triggered by the rapidly falling levels of erythropoietin seen after intravenous administration, and may be the normal mechanism for reduction of red cell mass in newborns. It may play a role in chronic diseases including malaria and sickle cell anemia. New erythropoietin products and methods of administration avoid the intermittent rapid decreases associated with the stimulus for neocytolysis, but study of this phenomenon may yield further improvements in drug design. PMID- 14754398 TI - Red blood cells as modulators of T cell growth and survival. AB - T cell homeostasis is largely controlled by a balance between cell death and survival and anomalies in either process account for a number of diseases linked to excessive or faulty T cell growth. Yet, the influence that cells outside the immunological system have on these processes has only recently received attention. Accumulated evidence indicate that homeostasis of the CD4+ and CD8+ T cell pools is highly dynamic and regulated by signals delivered by cells and molecules present in the different internal microenvironments. The major function of red blood cells (RBC) is generally considered to be oxygen and carbon dioxide transport. In recent years, however, RBC have been implicated in the regulation of basic physiological processes, from vascular contraction and platelet aggregation to T cell growth and survival. Regulation of T cell survival by RBC may influence the response of selected subsets of T cells to internal or external stimuli and may help explaining the immunomodulatory activities of red blood cells. By interfering in the balance between death and survival RBC become potential tools that can be manipulated to improve or reverse pathological situations characterized by anomalies in the control of T cell growth. PMID- 14754399 TI - Complement and complement regulatory proteins as potential molecular targets for vascular diseases. AB - By-products of complement activation and complement regulatory proteins are increasingly recognized to play an important pathogenic role in a variety of vascular diseases including atherosclerosis, ischemia and reperfusion injury, hyperacute graft rejection, vasculitis, and the vascular complications of human diabetes. "Self" damage by autologous complement is mediated by activation products of the complement cascades or by direct insertion of the membrane attack complex (MAC) into cell membranes. Specifically, insertion of MAC complexes into endothelial cells results in the release of an array of growth factors and cytokines that induces proliferation, inflammation and thrombosis in the vascular wall. This paper reviews complement and complement regulatory proteins with specific focus on the vasculature and vascular diseases; it highlights complement and its regulators as potential targets for the rational design of mechanism specific drugs for the treatment of some of the most prevalent human diseases. PMID- 14754400 TI - Antibody-mediated endothelial cell damage via nitric oxide. AB - Vascular disorders, resulting from endothelial cell dysfunction, may be caused by various stimuli, including infectious pathogens, cytotoxic reagents, and pathophysiological mechanisms mediated by immune responses. Endothelial cell dysfunction characterized by apoptosis and abnormal immune activation is, at least in part, induced by anti-endothelial cell antibody (AECA) in some cases of autoimmune disease. However, the molecular mechanisms of AECA-mediated pathogenetic damage to host vascular system remain unclear. The dual role of nitric oxide (NO) both in endothelial cell apoptosis and survival has been described. In this paper, endothelial cell apoptosis caused by the presence of cross-reactive AECA via a NO-mediated mechanism is demonstrated in dengue virus infection. Endothelial cells undergo apoptosis via the mitochondria-dependent pathway that is regulated by NO production. NO-regulated endothelial cell injury thus may play a role in the disruption of vessel endothelium and contribute to the AECA-induced pathogenesis of vasculopathy. The modulation of NO may provide the therapeutic strategies for autoimmune diseases by preventing the AECA mediated endothelial cell damage. PMID- 14754401 TI - Angiogenesis inhibitors: current & future directions. AB - The field of angiogenesis modulation is at a major crossroad. A tremendous advancement in basic science in this field is providing an excellent support for the concept, which is in contrast to a lack of strong clinical support to date. With regard to the large gap between experimental data and clinical data, the best model of human malignancy is in human cancer patients and the best model of human ocular angiogenesis-mediated disorders such as diabetic retinopathy (DR) and age related macular degeneration (AMD) is in human RD and AMD patients. Additionally, clinical outcomes should include benefit/risk ratios, hard end points (mortality and quality of life as opposed to increased microvascular density with pro-angiogenic agents or tumor size reduction with anti-angiogenesis agents) as well as cost effectiveness. Experimental models should be used to provide guidance, placebo effect, comparative data, and mechanistic understanding as opposed to being used for expected clinical efficacy. We also have to understand existing strategies and how angiogenesis modulation can add further value (i.e. not to replace existing strategy but rather improve efficacy/safety). Recent investigation defined numerous strategies in the modulation of angiogenesis. Those strategies are driven from haemostatic, fibrinolytic, cell adhesion molecules, extracellular matrix, growth factors, and other endogenous systems involved in the modulation of angiogenesis. PMID- 14754402 TI - Angiogenesis: a target for cancer therapy. AB - The induction of neoangiogenesis is a critical step already present at the early stages of tumor development and dissemination. The progressive identification of molecules playing a relevant role in neoangiogenesis has fostered the development of a wide variety of new selective agents. Antiangiogenic drugs should be integrated with conventional therapies; however, the design of the best sequence and timing for such combined treatments are still under investigation. In this review will be discussed the signal transduction mechanisms of angiogenic molecules, the development of specific inhibitors and their translation into clinical studies and, finally, the new perspectives in antiangiogenic therapy. PMID- 14754403 TI - Antiangiogenic therapy. AB - Angiogenesis, the formation of blood vessels from preexisting ones, plays a crucial role in tumor progression. Activation of an "angiogenic switch" allows tumor cells to invade and metastasize. The growing interest in the use of antiangiogenic agents in the treatment and prevention of cancer lies in the theoretical advantages of this molecularly targeted modality of chemotherapy. Delivery of antiangiogenic agents are not complicated by having to penetrate large bulky masses but, instead, have easy access to tumoral endothelial cells. Antiangiogenic drugs may not cause cytopenias and thus will avoid many of the unwarranted toxicities of standard chemotherapeutic agents. Because they act directly on nascent endothelial cells, antiangiogenic agents may avoid tumor resistance mechanisms. If antiangiogenic agents are successful, they might be applicable to many tumor types and not be dependent on cell type or growth fraction of cells within a tumor. However, several important obstacles remain with regards to using antiangiogenic drugs in clinical trials with which we must contend in order to determine accurately the efficacy of these agents. In this article, we review the different classes of antiangiogenic agents available, ongoing clinical trials, as well as potential pitfalls and future directions in this exciting field. PMID- 14754404 TI - The urokinase plasminogen activator system: role in malignancy. AB - The urokinase plasminogen activator (uPA) system consists of the serine protease uPA, its glycolipid-anchored receptor, uPAR and its 2 serpin inhibitors, plasminogen activator inhibitor-1 (PAI-1) and plasminogen activator inhibitor-2 (PAI-2). Recent findings suggest that the uPA system is causally involved at multiple steps in cancer progression. In particular, uPA has been implicated in remodelling of the extracellular matrix, enhancing both cell proliferation and migration and modulating cell adhesion. Consistent with its role in cancer progression, multiple groups have shown that high levels of uPA in primary breast cancers are independently associated with adverse outcome. Paradoxically, high levels of PAI-1 also correlate with poor prognosis in patients with breast cancer. The prognostic value of uPA/PAI-1 in axillary node-negative breast cancer patients was recently validated using both a prospective randomised trial and a pooled analysis, i.e., in 2 different Level 1 Evidence studies. Assay of uPA and PAI-1 may thus help identify low risk node-negative patients for whom adjuvant chemotherapy is unnecessary. Finally, preclinical studies show that either inhibition of uPA catalytic activity or prevention of uPA binding to its receptor reduces tumor growth, angiogenesis and metastasis. PMID- 14754405 TI - Resistance to anti-VEGF agents. AB - The number of anti-angiogenic agents developed for clinical use has risen greatly over the past decade. Currently, more than 80 are in trials ranging from phase I through to phase III studies and many more are in preclinical evaluation. Much hope was envisaged for these new agents to become the panacea of anti-tumoural treatment. Unfortunately the single agent activity to date has proven to be disappointing although one trial has recently reported a survival advantage when chemotherapy was administered with anti-VEGF antibodies in the setting of advanced colorectal cancer. To an extent, this may be due to great expectations of cytostatic compounds, but recently many factors have been examined to explain the differences between clinical and experimental findings. In this review, some of the factors responsible for the discrepancy are examined, with a specific focus on inhibitors of VEGF. The key factors responsible for the lack of activity are tumour heterogeneity and redundancy in the VEGF signalling system. An increased understanding of these factors is critical to the development of effective anti-angiogenic agents and need to be taken into account as new generations of drugs emerge. PMID- 14754406 TI - Molecular targeting of lymphatics for therapy. AB - The dysfunction or proliferation of lymphatic vessels (lymphangiogenesis) is linked to a number of pathological conditions including lymphedema and cancer. The recent discovery and characterisation of the lymphangiogenic growth factors vascular endothelial growth factor-C (VEGF-C) and VEGF-D and of their receptor on lymphatic endothelial cells, VEGFR-3, has provided an understanding of the molecular mechanisms controlling the growth of lymphatic vessels. In addition, other genes and protein markers have been identified with specificity for lymphatic endothelium that have enhanced the characterization and isolation of lymphatic endothelial cells. Our growing understanding of the molecules that control lymphangiogenesis allows us to design more specific drugs with which to manipulate the relevant signalling pathways. Modulating these pathways and other molecules with specificity to the lymphatic system could offer alternative treatments for a number of important clinical conditions. PMID- 14754407 TI - Development and engineering of lymphatic endothelial cells: clinical implications. AB - Studies on the lymphatic endothelium have been hampered by the difficulty to identify lymphatic endothelial cells (LECs) and to distinguish them from blood vascular endothelial cells (BECs). The situation was greatly improved by the identification of molecules with high specificity for LECs. A great deal of progress in the field of lymphangiogenesis research has been due to the detection of lymphangiogenic growth factors and their receptors, and there is growing evidence that these molecules are also involved in tumor-induced lymphangiogenesis and lymphatic dissemination of tumor cells. There is a considerable spectrum of congenital and acquired lymphedema-lymphangiodysplasia syndromes ranging from primary aplasia, hypoplasia and hyperplasia to secondary (acquired) obstructive, obliterative and surgical hindrance of lymph drainage. Consequently, there are a number of clinical applications for therapeutics that either inhibit or induce lymphangiogenesis. Although natural lymphatic regeneration is mostly very efficient, engineering of LECs may be useful in cases of lymphatic aplasia or hypoplasia. To achieve these goals, studies on the embryonic development and differentiation of LECs will reveal the key regulatory factors that need to be targeted. PMID- 14754409 TI - Drug binding domains of MRP1 (ABCC1) as revealed by photoaffinity labeling. AB - Drug resistance is a major impediment in the treatment of cancer patients receiving single or multiple drug treatment. Efforts to reverse drug resistance of tumor cells have not been successful. In recent years, considerable emphasis has been placed on understanding the underlying mechanisms that confer drug resistance. The expression of the multidrug resistance protein 1 (MRP1 or ABCC1) in cancer cells has been shown to confer resistance to diverse classes of anti cancer drugs. MRP1 is a member of the ATP-binding cassette (ABC) family whose function, in tumor cells, is to reduce drug accumulation through energized drug efflux. To learn more about the functions of MRP1 in tumor drug resistance, knowledge of the protein binding characteristics and the location of its binding sites are essential. Photoaffinity labeling (PAL) has emerged as a leading technique that can rapidly shed light on a protein's drug binding characteristics and ultimately drug binding domains. Several MRP1-specific photoreactive probes have been developed. PAL of MRP1 was first demonstrated with the quinoline-based drug, IAAQ. Other studies showed that the high affinity endogenous substrate of MRP1, LTC(4), has intrinsic photoreactive properties and binds within both N- and C-terminal domains of MRP1. LTC(4) is conjugated to glutathione (GSH), a property common to several MRP1 substrates. In addition, several unconjugated drugs have been identified that interact with MRP1: [(3)H]VF-13,159, IAAQ, IACI and IAARh123. Mapping studies showed that IACI and IAARh123 bind two sites within transmembrane (TM) regions 10-11 and 16-17 of MRP1. Interestingly, the GSH dependent PAL of [(125)I]azidoAG-A and [(125)I]LY475776 occurs within, or proximal to TM 16-17. The PAL with several analogs of GSH, IAAGSH and azidophenacyl-[(35)S]GSH found to interact specifically with MRP1 within TM 10-11 and TM 16-17 in addition to binding two cytoplasmic regions in MRP1, L0 and L1. This review focuses on the use of PAL for studying MRP1 interactions with various drugs and cell metabolites. Furthermore, knowledge of MRP1 drug binding domains, as identified by PAL with various photoreactive drug analogs, provides an important first step towards more detailed analyses of MRP1 binding domains. PMID- 14754411 TI - Reversing agents for ATP-binding cassette (ABC) transporters: application in modulating multidrug resistance (MDR). AB - One of the main reasons for the failure in cancer chemotherapy is the existence of multidrug resistance (MDR) mechanisms. One form of MDR phenotype is contributed by a group of plasma membrane proteins that belong to a large superfamily of proteins called the ATP-binding cassette (ABC) transporters. There has been intense search for compounds, which can act at reversing MDR phenotype exhibited by ABC transporters such as P-glycoprotein (P-gp), multidrug-resistance protein (MRP) and breast cancer resistance protein (BCRP). Reversing agents can be designed to target MDR-associated ABC transporters at three levels - the protein, mRNA or DNA level. This review aims at describing, over-viewing and discussing currently known MDR reversing agents, which have been shown to act at either of the three levels against ABC transporters. Other potential agents and strategies, which can be used to reverse the MDR phenotype, are also discussed. PMID- 14754410 TI - Multidrug resistance in cancer chemotherapy and xenobiotic protection mediated by the half ATP-binding cassette transporter ABCG2. AB - ABCG2, also termed BCRP/MXR/ABCP, is a half ATP-binding cassette (ABC) transporter expressed on plasma membranes. ABCG2 was independently cloned from placenta as well as cell lines selected for resistance to mitoxantrone or anthracyclines. ABCG2 consists of a nucleotide-binding domain (NBD) at the amino terminus and a transmembrane domain (TMD) at the carboxyl terminus and it is postulated to form a homodimer to perform its biological functions. Over expression of ABCG2 in cell lines confers resistance on a wide variety of anticancer drugs including mitoxantrone, daunorubicin, doxorubicin, topotecan and epirubicin. The expression of ABCG2 has been implicated in multidrug resistance (MDR) of acute myeloid leukemia and some solid tumors. In addition, ABCG2 can transport several fluorescent dyes or toxins. ABCG2 is found to be expressed in epithelial cells of intestine and colon, liver canaliculi, and renal tubules, where it serves to eliminate the plasma level of orally administered anticancer drugs as well as ingested toxins. ABCG2 is found to be highly expressed in placenta and the luminal surface of microvessel endothelium blood-brain barrier where it may play a role in limiting the penetration of drugs, such as topotecan from the maternal plasma into the fetus and from blood to brain. A variety of inhibitors for ABCG2 including GF120918 may prove useful for sensitizing cancer cells to chemotherapy or altering the distribution of orally administered drug substrates of ABCG2. Interestingly, ABCG2 is also expressed highly in hematopoietic stem cells. However, the function of ABCG2 in stem cells is currently unknown, although it may provide protection to stem cells from a variety of xenobiotics. PMID- 14754412 TI - Structure of multidrug-resistance proteins of the ATP-binding cassette (ABC) superfamily. AB - Multidrug resistance of tumors, characterized by resistance against a variety of chemically unrelated anticancer agents, can be caused by overexpression of ATP binding cassette (ABC) proteins, such as P-glycoprotein and MRP1. These multidrug resistance proteins are plasma-membrane proteins that actively extrude chemotherapeutic agents from the cell interior, decreasing drug accumulation and thus, allowing the cells to survive in the presence of toxic levels of anticancer agents. ABC proteins contain multispanning transmembrane domains and nucleotide binding domains (NBDs). The NBDs are responsible for the ATP binding/hydrolysis that drives drug transport, and their structure is conserved independently of the degree of primary-sequence homology. The transmembrane domains contain the drug binding sites that are likely located in a flexible internal chamber that is sufficiently large to accommodate different drugs. It has been recently proposed that dimerization of the NBDs induced by ATP binding is a key step for the coupling of ATP hydrolysis to substrate transport. The power stroke for substrate transport can be the formation or the dissociation of the dimers. Since the NBDs and TMDs are tightly associated, association/dissociation of the NBDs may control the "gate" of the translocation pathway, formed by intracellular loops. In the case of P-glycoprotein it seems that the power stroke for transport is ATP binding (and therefore NBD dimerization), and not hydrolysis, because the major conformational and functional changes seem to occur at this step. PMID- 14754408 TI - Identification and characterization of the binding sites of P-glycoprotein for multidrug resistance-related drugs and modulators. AB - A major problem in cancer treatment is the development of resistance to multiple chemotherapeutic agents in tumor cells. A major mechanism of this multidrug resistance (MDR) is overexpression of the MDR1 product P-glycoprotein, known to bind to and transport a wide variety of agents. This review concentrates on the progress made toward understanding the role of this protein in MDR, identifying and characterizing the drug binding sites of P-glycoprotein, and modulating MDR by P-glycoprotein-specific inhibitors. Since our initial discovery that P glycoprotein binds to vinblastine photoaffinity analogs, many P-glycoprotein specific photoaffinity analogs have been developed and used to identify the particular domains of P-glycoprotein capable of interacting with these analogs and other P-glycoprotein substrates. Furthermore, significant advances have been made in delineating the drug binding sites of this protein by studying mutant P glycoprotein. Photoaffinity labeling experiments and the use of site-directed antibodies to several domains of this protein have allowed the localization of the general binding domains of some of the cytotoxic agents and MDR modulators on P-glycoprotein. Moreover, site-directed mutagenesis studies have identified the amino acids critical for the binding of some of these agents to P-glycoprotein. Furthermore, equilibrium binding assays using plasma membranes from MDR cells and radioactive drugs have aided our understanding of the modes of drug interactions with P-glycoprotein. Based on the available data, a topological model of P glycoprotein and the approximate locations of its drug binding sites, as well as a proposed classification of multiple drug binding sites of this protein, is presented in this review. PMID- 14754413 TI - Cationic lipid-DNA complexes for gene therapy: understanding the relationship between complex structure and gene delivery pathways at the molecular level. AB - Cationic liposomes (CLs) are used as gene vectors (carriers) in worldwide human clinical trials of non-viral gene therapy. These lipid-gene complexes have the potential of transferring large pieces of DNA of up to 1 million base-pairs into cells. As our understanding of the mechanisms of action of CL-DNA complexes remains poor, transfection efficiencies are still low when compared to gene delivery with viral vectors. We describe recent studies with a combination of techniques (synchrotron x-ray diffraction for structure determination, laser scanning confocal microscopy to probe the interactions of CL-DNA particles with cells, and luciferase reporter-gene expression assays to measure transfection efficiencies in mammalian cells), which collectively are beginning to unravel the relationship between the distinctly structured CL-DNA complexes and their transfection efficiency. The work described here is applicable to transfection optimization in ex vivo cell transfection, where cells are removed and returned to patients after transfection. CL-DNA complexes primarily form a multilayered sandwich structure with DNA layered between the cationic lipids (labeled L(alpha)(C)). On rare occasions, an inverted hexagonal structure with DNA encapsulated in lipid tubules (labeled H(II)(C)) is observed. A major recent insight is that for L(alpha)(C) complexes the membrane charge density sigma(M) of the CL-vector, rather than the charge of the cationic lipid alone, is a key universal parameter that governs the transfection efficiency of L(alpha)(C) complexes in cells. The parameter sigma(M) is a measure of the average charge per unit area of the membrane, thus taking into account the amount of neutral lipids. In contrast to L(alpha)(C) complexes, H(II)(C) complexes containing the lipid 1,2 dioleoyl-sn-glycerophosphatidylethanolamine (DOPE) exhibit no dependence on sigma(M). The current limiting factor to transfection by cationic lipid vectors appears to be the tight association of a fraction of the delivered exogenous DNA with cationic cellular molecules, which may prevent optimal transcriptional activity. Future directions are outlined, which make use of surface functionalized CL-DNA complexes suitable for transfection in vivo. PMID- 14754414 TI - Modeling of cationic lipid-DNA complexes. AB - Cationic lipid-DNA complexes, often referred to as lipoplexes, are formed spontaneously in aqueous solutions upon mixing DNA and liposomes composed of cationic and nonionic lipids. Understanding the mechanisms underlying lipoplex formation, structure and phase behavior is crucial for their further development and design as non-viral transfection vectors in gene therapy. From a physical point of view, lipoplexes are ordered, self-assembled, composite aggregates. Their preferred spatial geometry and phase behavior are governed by a delicate coupling between the electrostatic interactions which drive lipoplex formation and the elastic properties of the constituent lipid layers, both depending on the molecular nature and composition of the lipid mixture. In this review we outline some recent efforts to model the microscopic structure, energetic and phase behavior of cationic lipid-DNA mixtures, focusing on the two principal aggregation geometries: the lamellar (L(alpha)(C)), or "sandwich" complexes, and the hexagonal (H(II)(C)), or "honeycomb" complexes. We relate the structural and thermodynamic properties of these two "canonical" lipoplex morphologies to their appearance in phase diagrams of DNA-lipid mixtures, emphasizing the crucial role fulfilled by the molecular packing characteristics of the cationic and neutral lipids, as reflected in the curvature elastic properties of the mixed lipid layer. PMID- 14754415 TI - Conception and realization of a non-cationic non-viral DNA vector. AB - Cationic non-viral DNA vectors are very successful in in vitro transfections but less efficient in in vivo tests. This seems mainly due to the cationic nature of the molecules used to complex DNA. In this article, we describe the design and the route towards the realization of a non-viral non-cationic vector. The strategy follows three steps: first, the incorporation of DNA to a lamellar phase; second, the making of multilamellar vesicles containing a high loading of DNA by shearing the lamellar phase and, finally, the grafting of peptides onto the surface of the vesicles to target a specific receptor on the cells. Throughout this process, we had to overcome many obstacles; this review describes the present state of our work and summarizes the remaining steps. PMID- 14754416 TI - Self-assembling nucleic acid delivery vehicles via linear, water-soluble, cyclodextrin-containing polymers. AB - Non-viral (synthetic) nucleic acid delivery systems have the potential to provide for the practical application of nucleic acid-based therapeutics. We have designed and prepared a tunable, non-viral nucleic acid delivery system that self assembles with nucleic acids and centers around a new class of polymeric materials; namely, linear, water-soluble cyclodextrin-containing polymers. The relationships between polymer structure and gene delivery are illustrated, and the roles of the cyclodextrin moieties for minimizing toxicity and forming inclusion complexes in the self-assembly processes are highlighted. This vehicle is the first example of a polymer-based gene delivery system formed entirely by self-assembly. PMID- 14754417 TI - The vesosome-- a multicompartment drug delivery vehicle. AB - Assembling structures to divide space controllably and spontaneously into subunits at the nanometer scale is a significant challenge, although one that biology has solved in two distinct ways: prokaryotes and eukaryotes. Prokaryotes have a single compartment delimited by one or more lipid-protein membranes. Eukaryotes have nested-membrane structures that provide internal compartments- such as the cell nucleus and cell organelles in which specialized functions are carried out. We have developed a simple method of creating nested bilayer compartments in vitro via the "interdigitated" bilayer phase formed by adding ethanol to a variety of saturated phospholipids. At temperatures below the gel liquid crystalline transition, T(m), the interdigitated lipid-ethanol sheets are rigid and flat; when the temperature is raised above T(m), the sheets become flexible and close on themselves and the surrounding solution to form closed compartments. During this closure, the sheets can entrap other vesicles, biological macromolecules, or colloidal particles. The result is efficient and spontaneous encapsulation without disruption of even fragile materials to form biomimetic nano-environments for possible use in drug delivery, colloidal stabilization, or as microreactors. The vesosome structure can take full advantage of the 40 years of progress in liposome development including steric stabilization, pH loading of drugs, and intrinsic biocompatibility. However, the multiple compartments of the vesosome give better protection to the interior contents in serum, leading to extended release of model compounds in comparison to unilamellar liposomes. PMID- 14754418 TI - The Vif protein of human immunodeficiency virus type 1 (HIV-1): enigmas and solutions. AB - HIV-1 and other complex retroviruses express six auxiliary genes in addition to the canonical retroviral genes, gag, pol and env. Vif (virion infectivity factor) protein is absolutely essential for productive HIV-1 infection of peripheral blood lymphocytes and macrophages, the two major HIV-1 target cells in vivo. However, Vif is not required for production of infectious particles in several human cell lines. In spite of the prominent phenotype of Vif mutations, the mechanism of its action remains unknown. During the last decade several models were suggested to explain the mechanism of Vif activity. One view holds that Vif is active in virions after budding or after entry into target cells during the early stages of HIV-1 replications. The second view places the action of Vif at the late stage of HIV-1 replication in virus producing cells, which affects the production of infectious virus. According to this view, Vif either compensates the cell factor required for production of infectious virus, or alternatively, it neutralizes a cell factor, which prevents the production of infectious particles in these cells. This review is addressed to summarize the models envisioned to explain Vif activities. The findings described here, that Vif interacts with viral and cellular components, elaborates the importance of Vif as a novel target for developing anti HIV-1 drugs. PMID- 14754419 TI - Revisiting pitfalls, problems and tentative solutions for assaying mitochondrial respiratory chain complex III in human samples. AB - The assessment of mitochondrial respiratory chain enzyme activity in human samples is a difficult task due to both the small amount of tissue generally available and the frequent need to perform enzyme activity measurement in crude mitochondrial fraction. This is particularly true for the measurement of complex III activity which partial deficiency can be easily overlooked. In this review, we first consider the several interfering reactions occurring when measuring this activity. We subsequently describe the use of an alkyl glycoside detergent, lauryl maltoside, to keep these interfering reactions to a very low level. Next, we quantify the effect of the detergent on the actual measurement of complex III in various human tissue samples and cells. Finally, we also demonstrate that the use of the detergent allows (i) a better detection of an inherited partial defect affecting cytochrome b, a catalytic subunit of the mitochondrial complex III, (ii) to possibly discriminate decreased complex III activity resulting from an abnormal complex III assembly (BCS1 gene mutation) from an hampered catalytic activity originating from a cytochrome b mutation. This detailed review of the problems associated with complex III assessment and of their tentative solution highlights the difficulties still encountered in the measurements of mitochondrial respiratory chain in humans. PMID- 14754420 TI - Alpha-galactosylceramide: potential immunomodulatory activity and future application. AB - alpha-Galactosylceramide (alpha-GalCer), is a glycolipid which has been identified as a ligand recognized by a special group of immune T cells, known as invariant NKT cells. alpha-GalCer can powerfully activate invariant NKT cells to produce immunoregulatory cytokines, including interferon-gamma and IL-4, and thereby exert a variety of subsequent effects on other cells in the immune system. Recent studies have revealed the mechanism of alpha-GalCer-induced iNKT cell-activation in immune responses to tumors and microbes, and in the suppression of autoimmune diseases. In this review, we discuss the potential immunomodulatory activity of alpha-GalCer and its possible future application for clinical studies in humans. PMID- 14754421 TI - Effects of the capsular polysaccharides of Cryptococcus neoformans on phagocyte migration and inflammatory mediators. AB - An important virulence factor of the pathogenic fungus Cryptococcus neoformans is its polysaccharide capsule. The capsular polysaccharides glucuronoxylomannan (GXM), galactoxylomannan (GalXM) and the mannoproteins (MPs) display various immunomodulatory effects on the host response, such as the inhibition of phagocytosis, suppression of T-cell mediated immunity, and induction of immunogenic tolerance. Moreover, these capsular polysaccharides are able to interfere with the migration of phagocytes despite adequate stimulation of chemokine production and their concerted action accounts for the mild inflammatory response often observed in cryptococcosis. Different mechanisms contribute to this phenomenon. First, cryptococcal polysaccharides impair leukocyte migration towards chemoattractants. A combination of the intrinsic chemoattracting properties of circulating polysaccharides and the ability to induce cross-desensitization of chemokine receptors prevents leukocytes from leaving the bloodstream and migrating towards inflammatory site. Polysaccharide induced repressive effects on the C5a receptor expression on neutrophils may also add to this impaired chemokinesis. Second, polysaccharides interfere with leukocyte adhesion to and migration through the endothelium. Both GXM and MP-4 induce L-selectin shedding from the surface of leukocytes; hence, interference with leukocyte rolling on the endothelium can be expected. GXM also interferes with the subsequent process of firm leukocyte adhesion to the endothelium in vitro. Thirdly, capsular polysaccharides enhance the production of anti inflammatory interleukin-10 (IL-10) and induce tumor necrosis factor-alpha (TNFalpha) receptor loss from the surface of neutrophils. The capacity to reduce neutrophil influx makes cryptococcal polysaccharides interesting compounds to study in clinical models of inflammation (i.e.; sepsis, auto-immune disorders) in which leukocyte influx can be potentially damaging to host tissues. PMID- 14754423 TI - Pharmacological approaches in the treatment of atrial fibrillation. AB - Atrial fibrillation (AF) is the most common sustained cardiac arrhythmia and is associated with substantial cardiovascular morbidity and mortality. The arrhythmia can be initiated and/or maintained by rapidly firing foci, single- and multiple-circuit reentry. Once initiated, AF alters atrial electrical and structural properties (atrial remodeling) in a way that promotes its own maintenance and recurrence and may alter the response to antiarrhythmic drugs. Thus, initial episodes of paroxysmal (self-terminating) AF lengthens to the point where the arrhythmia becomes persistent (requires cardioversion to restore sinus rhythm) and permanent. AF usually requires a trigger for initiation and a favorable electrophysiological and/or anatomical substrate for maintenance. The substrate includes both cardiovascular (coronary artery disease, valvular heart disease, heart failure, hypertension, dilated cardiomyopathy) and non cardiovascular diseases (thyrotoxicosis, pulmonary diseases). Accordingly, the initial step in patients with AF requires a careful assessment of symptoms and identification of underlying reversible triggers and potentially modifiable underlying structural substrate and treat them aggressively. In contrast to other cardiac arrhythmias, antiarrhythmic drugs (ADs) are the mainstay of therapy. Long term treatment of AF is directed to restore and maintain the sinus rhythm with class I and III ADs (rhythm-control) or to allow AF to persist and ensure that the ventricular rate is controlled (rate-control) with atrioventricular nodal blocking drugs (digoxin, beta-blockers, verapamil, diltiazem) and prevent thromboembolic complications with anticoagulants. However, the long-term efficacy of ADs for preventing AF recurrence is far from ideal, because of limited efficacy (AF recurs in at least one-half of the patients) and potential side effects, particularly proarrhythmia. Thus, the choice of the appropriate AD will depend on the temporal pattern of the arrhythmia, the presence of associated diseases, easy of administration and adverse effects profile, particularly the risk of proarrhythmia. The recent finding that angiotensin converting enzyme inhibitors and beta-blockers reduce the incidence of AF in patients post myocardial infarction with left ventricular dysfunction confirmed the importance of targeting the underlying arrhythmogenic substrate. This review focuses on the mechanisms underlying AF and the mechanism of action and the efficacy and safety profile of the ADs used in the treatment of atrial fibrillation. The advantages and disadvantages of rhythm and rate control, the role pill in a pocket concept and the role of the new ADs are dicussed. PMID- 14754422 TI - Theoretical possibilities for the development of novel antiarrhythmic drugs. AB - One possible mechanism of action of the available K-channel blocking agents used to treat arrhythmias is to selectively inhibit the HERG plus MIRP channels, which carry the rapid delayed rectifier outward potassium current (I(Kr)). These antiarrhythmics, like sotalol, dofetilide and ibutilide, have been classified as Class III antiarrhythmics. However, in addition to their beneficial effect, they substantially lengthen ventricular repolarization in a reverse-rate dependent manner. This latter effect, in certain situations, can result in life-threatening polymorphic ventricular tachycardia (torsades de pointes). Selective blockers (chromanol 293B, HMR-1556, L-735,821) of the KvLQT1 plus minK channel, which carriy the slow delayed rectifier potassium current (I(Ks)), were also considered to treat arrhythmias, including atrial fibrillation (AF). However, I(Ks) activates slowly and at a more positive voltage than the plateau of the action potential, therefore it remains uncertain how inhibition of this current would result in a therapeutically meaningful repolarization lengthening. The transient outward potassium current (I(to)), which flows through the Kv 4.3 and Kv 4.2 channels, is relatively large in the atrial cells, which suggests that inhibition of this current may cause substantial prolongation of repolarization predominantly in the atria. Although it was reported that some antiarrhythmic drugs (quinidine, disopyramide, flecainide, propafenone, tedisamil) inhibit I(to), no specific blockers for I(to) are currently available. Similarly, no specific inhibitors for the Kir 2.1, 2.2, 2.3 channels, which carry the inward rectifier potassium current (I(kl)), have been developed making difficult to judge the possible beneficial effects of such drugs in both ventricular arrhythmias and AF. Recently, a specific potassium channel (Kv 1.5 channel) has been described in human atrium, which carries the ultrarapid, delayed rectifier potassium current (I(Kur)). The presence of this current has not been observed in the ventricular muscle, which raises the possibility that by specific inhibition of this channel, atrial repolarization can be lengthened without similar effect in the ventricle. Therefore, AF could be terminated and torsades de pointes arrhythmia avoided. Several compounds were reported to inhibit I(Kur)(flecainide, tedisamil, perhexiline, quinidine, ambasilide, AVE 0118), but none of them can be considered as specific for Kv 1.5 channels. Similarly to Kv 1.5 channels, acetylcholine activated potassium channels carry repolarizing current (I(KAch)) in the atria and not in the ventricle during normal vagal tone and after parasympathetic activation. Specific blockers of I(KAch) can, therefore, also be a possible candidate to treat AF without imposing proarrhythmic risk on the ventricle. At present several compounds (amiodarone, dronedarone, aprindine, pirmenol, SD 3212) were shown to inhibit I(KAch) but none of them proved to be selective. Further research is needed to develop specific K-channel blockers, such as I(Kur)and I(KAch) inhibitors, and to establish their possible therapeutic value. PMID- 14754424 TI - Pharmacological modulation of I(Ks): potential for antiarrhythmic therapy. AB - The slowly (I(Ks)) and rapidly (I(Kr)) activating delayed rectifier K(+) currents play important roles in cardiac ventricular repolarization. Compared with I(Kr), however, I(Ks) has important distinguishing characteristics, including beta adrenergic receptor stimulation and accumulation at rapid rates that may impart significant therapeutic relevance. Therefore, development of selective I(Ks) inhibitors has been pursued as a strategy for providing potentially safer and more effective Class III antiarrhythmic agents and pharmacological tools for elucidating the normal physiological and potential pathological role of I(Ks) in cardiac repolarization. We have identified a series of 3-Acylamino-1,4 benzodiazepines that are very potent and selective inhibitors of I(Ks). A representative compound, L-768,673 (1) (IC(50)~8nM), has been extensively characterized for its pharmacologic activity. L-768,673 concentration-dependently prolongs action potential duration in a frequency-independent manner in vitro, but decreases transmural dispersion of refractoriness, a risk factor for arrhythmia induction. In conscious dogs, L 768,673 administered IV (0.3-100 micro g/kg) and PO (0.03-1 mg/kg) elicits consistent but limited (5-15%) QT(c) prolongation, and increases ventricular refractory period more at fast than at slow pacing rates, indicating a "forward" rate-dependence in vivo. In an anesthetized canine model of anterior myocardial infarction, I(Ks) blockers suppress the development of ischemic ventricular fibrillation at intravenous doses that minimally prolong the QT interval. I(Ks) blockers display an interesting and intriguing profile of effects on cardiac electrophysiologic parameters that differ in remarkable ways from other selective Class III agents such as I(Kr) blockers. It remains to be determined if these properties can be exploited clinically to provide more effective and safer treatment of cardiac arrhythmias. PMID- 14754425 TI - Profile of I(Ks) during the action potential questions the therapeutic value of I(Ks) blockade. AB - The goal of this paper is two fold. First, we attempt to review the reports available on the role of I(Ks) in myocardial repolarization. Based on theoretical considerations and experimental results, it seems reasonable to assume that I(Ks)blockade will lengthen the action potential. However, results obtained with I(Ks) blockers, like chromanol 293B or L-735,821, are conflicting, since from slight lengthening to marked prolongation of action potentials were equally obtained. Although these contradictory results were explained by interspecies or regional differences, the role of I(Ks) in repolarization is a matter of growing dispute. In the second part of this study, we simulated the performance of I(Ks) during cardiac action potentials. We compared the profile of the predicted current in three mathematical models in order to determine the relative role of the current in repolarization. We studied the effect of the cycle length, action potential duration and height of the plateau on the profile of I(Ks) in epicardiac, endocardiac and midmyocardiac ventricular action potentials. The results indicate that the height of the plateau is the most important parameter to control activation of I(Ks)in cardiac tissues, and accordingly, the interspecies and regional differences observed in the efficacy of I(Ks) blockers are likely due to the known differences in action potential morphology. We conclude also that I(Ks)blockade may have unpredictable effects on the length of the action potential in a diseased heart, questioning the possible therapeutic value of drugs blocking I(Ks). PMID- 14754426 TI - Novel antiarrhythmic compounds with combined class IB and class III mode of action. AB - Cardiac arrhythmias represent a major area of cardiovascular research, and for drug therapy, a large choice of antiarrhythmic agents have been available. However, clinical trials with antiarrhythmic drugs have recently indicated that serious side effects may considerably limit the use of various antiarrhythmic agents, in particular, for preventing arrhythmia-related mortality. Amiodarone with its complex mode of action, while exerting a strong and favorable antiarrhythmic action, posseses extracardiac untoward side effects originating from its chemical structure. In this paper, we report on our attempt to develop conceptually new, therapeutically valuable antiarrhythmic compounds, in which Class I/B and Class III features were combined into single molecules bearing no structural resemblance to amiodarone. Synthesis and pharmacological screening of series of N-(phenylalkyl)-N-(phenoxyalkyl)amines led us to discover some new promising compounds with the required dual mode of action. GYKI-16638, selected for further investigation, was also found to possess a remarkable in vivo antiarrhythmic effect, and it is now considered as a safe new antiarrhythmic drug candidate. PMID- 14754427 TI - Predicting molecular interactions in silico: I. A guide to pharmacophore identification and its applications to drug design. AB - A major goal in contemporary drug design is to develop new ligands with high affinity of binding toward a given protein receptor. Pharmacophore, which is the three-dimensional arrangement of essential features that enable a molecule to exert a particular biological effect, is a very useful model for achieving this goal. If the three dimensional structure of the receptor is known, pharmacophore is a complementary tool to standard techniques, such as docking. However, frequently the structure of the receptor protein is unknown and only a set of ligands together with their measured binding affinities towards the receptor is available. In such a case, a pharmacophore based strategy is one of the few applicable tools. Here we present a broad, yet concise guide to pharmacophore identification and review a sample of applications for drug design. In particular, we present the framework of the algorithms, classify their modules and point out their advantages and challenges. PMID- 14754428 TI - Predicting molecular interactions in silico: II. Protein-protein and protein-drug docking. AB - This article reviews the docking field starting from basic docking algorithms and describing the latest advances. We present the algorithmic framework and classify the state of-the-art methods. We point out the bottlenecks of the methods, like flexibility, absence of absolute scoring functions and explain what types of information can potentially be added to improve the results. PMID- 14754429 TI - A review of HIV-1 resistance to the nucleoside and nucleotide inhibitors. AB - The nucleoside reverse transcriptase inhibitors (NRTIs) were the first class of agents used for the treatment of HIV and remain an important component of combination antiretroviral therapy. Resistance to the NRTIs occurs by the acquisition of mutations in the reverse transcriptase gene that result in a structural change that either decreases the NRTI incorporation into the extending nucleotide chain or enhances removal of the NRTI from the terminated chain, also known as primer unblocking or pyrophosphorylysis. There are several major genetic mutational patterns of resistance and cross-resistance that evolve with the NRTIs including the thymidine analog mutations M41L, D67N, K70R, L210W, T215Y, and K219Q/E/W, the non-thymidine mutations M184V, L74V, and K65R, and the multidrug resistant Q151M complex, as well as others. Increasing knowledge of resistance and cross-resistance patterns that evolve on the NRTIs as well as the other antiretroviral classes will help optimize antiretroviral treatment strategies. Advancing knowledge of the biochemical and structural basis of resistance will aid in the design of newer compounds that are active against HIV resistant to the currently available drugs, ultimately prolonging virologic suppression and life in the millions of people who are infected with HIV. PMID- 14754430 TI - Resistance to HIV-1 entry inhibitors. AB - Resistance-testing technology has been incorporated into the standard of care for human immunodeficiency virus type 1 (HIV-1) infection and therapy with protease and reverse transcriptase inhibitors. Inhibitors of HIV-1 entry represent an emerging mode of antiretroviral therapy, and HIV-1 entry inhibitors encompass three mechanistically distinct classes of agents known as attachment inhibitors, coreceptor inhibitors, and fusion inhibitors. Each class of agent has demonstrated promise in controlled clinical trials, and understanding the determinants and evolution of viral resistance will be critical for the optimal development and deployment of these new treatment classes. Advances in resistance testing technologies have paralleled the development of HIV-1 entry inhibitor therapies, and the available data support the notion that attachment, coreceptor and fusion inhibitors offer complementary modes of therapy and distinct resistance profiles. PMID- 14754431 TI - Is resistance futile? AB - A global effort has been undertaken to control human immunodeficiency virus (HIV) though the development of vaccines and pharmacologics. Current FDA approved pharmacological inhibitors target two of the three viral enzymes critical to replication and maturation of infectious viral particles: reverse transcriptase (RT) and protease (Pr). Although combination therapies targeting RT and Pr have significantly reduced AIDS related morbidity and mortality, resistance to individual inhibitors is a growing concern. Currently, there are six protease inhibitors in clinical use. These inhibitors target the active site of protease using peptidomimetic transition state analogs based on natural substrates. However, treatment failures arise as a lack of compliance due to HIV-inhibitor pharmacokinetics, toxicity, and tolerance. This allows reduced HIV-inhibitor pressure, increased viral replication, and the emergence of drug resistant mutations. Continued use of protease inhibitors in the face of incomplete viral suppression may result in HIV-1 escape mutants not only being resistant to the protease inhibitor used, but to all clinically available protease inhibitors. Thus, new broad-based protease inhibitors are needed to control the emerging multi-drug, cross-resistant HIV-1. Moreover, given the emergence of cross resistant HIV-1, there is a need to target novel protease structural sites to reduce the risk of multi-drug cross-resistance. In this review, we discuss the resistance to protease inhibitors and the rationale for new strategies towards drug design for suppressing protease activity. We focus on the structure and function relationship and the influence that drug resistance mutants exert on the evolution of HIV-1 protease. PMID- 14754432 TI - Structural and thermodynamic basis of resistance to HIV-1 protease inhibition: implications for inhibitor design. AB - One of the most serious side effects associated with the therapy of HIV-1 infection is the appearance of viral strains that exhibit resistance to protease inhibitors. At the molecular level, resistance to protease inhibition predominantly takes the form of mutations within the protease molecule that preferentially lower the affinity of protease inhibitors with respect to protease substrates, while still maintaining a viable catalytic activity. Mutations associated with drug resistance occur within the active site cavity as well as distal sites. Active site mutations affect directly inhibitor/protease interactions while non-active site mutations affect inhibitor binding through long range cooperative perturbations. The effects of mutations associated with drug resistance are compounded by the presence of naturally occurring polymorphisms, especially those observed in non-B subtypes of HIV-1. The binding thermodynamics of all clinical inhibitors against the wild type protease, drug resistant mutations and non-B subtype HIV-1 proteases has been determined by high sensitivity isothermal titration calorimetry. In conjunction with structural information, these data have provided a precise characterization of the binding mechanism of different inhibitors and their response to mutations. Inhibitors that exhibit extremely high affinity and low susceptibility to the effects of mutations share common features and binding determinants even if they belong to different chemical scaffolds. These binding determinants define a set of rules and constraints for the design of better HIV-1 protease inhibitors. PMID- 14754433 TI - Mathematical approaches in the study of viral kinetics and drug resistance in HIV 1 infection. AB - We review some crucial aspects of drug therapy and viral resistance that have been investigated within the framework developed for the modelling of virus kinetics. First, we give a general overview on the use of mathematical models in the field of HIV research. We seek to identify the factors that determine the steady state virus load and show that stable reductions during antiviral therapy are difficult to explain within the standard model of virus dynamics. We discuss possible extensions that enable the models to account for the moderately reduced virus loads during non-suppressive treatment and argue that the residual viremia under suppressive treatment can probably be attributed to the survival of long lived infected cells, rather than to new rounds of replication. Next, we address the emergence of resistance during suppressive therapy and demonstrate that the resistant virus is more likely to be present already at the start of treatment than to be generated during therapy. The appearance of resistance after a prolonged period of initial suppression indicates that drug efficacy is not continuously maintained over time. We investigate the potential risks and benefits of therapy interruptions. Considering the effect of recombination, we argue that it probably decelerates, rather than accelerates the evolution of multidrug-resistant virus. We also review state-of-the-art methods for the estimation of fitness, which is crucial to the understanding of the emergence of resistance during therapy or the re-emergence of wild type upon the cessation of therapy. PMID- 14754434 TI - Predicting the impact of antiretrovirals in resource-poor settings: preventing HIV infections whilst controlling drug resistance. AB - There is currently an opportunity to carefully plan the implementation of antiretroviral (ARV) therapy in the developing world. Here, we use mathematical models to predict the potential impact that low to moderate usage rates of ARVs might have in developing countries. We use our models to predict the relationship between the specific usage rate of ARVs (in terms of the percentage of those infected with HIV who receive such treatment) and: (i) the prevalence of drug resistant HIV that will arise, (ii) the future transmission rate of drug resistant strains of HIV, and (iii) the cumulative number of HIV infections that will be prevented through more widespread use of ARVs. We also review the current state of HIV/AIDS treatment programs in resource-poor settings and identify the essential elements of a successful treatment project, noting that one key element is integration with a strong prevention program. We apply both program experience from Haiti and Brazil and the insights gleaned from our modeling to address the emerging debate regarding the increased availability of ARVs in developing countries. Finally, we show how mathematical models can be used as tools for designing robust health policies for implementing ARVs in developing countries. Our results demonstrate that designing optimal ARV-based strategies to control HIV epidemics is extremely complex, as increasing ARV usage has both beneficial and detrimental epidemic-level effects. Control strategies should be based upon the overall impact on the epidemic and not simply upon the impact ARVs will have on the transmission and/or prevalence of ARV-resistant strains. PMID- 14754435 TI - Fitness variations and their impact on the evolution of antiretroviral drug resistance. AB - The human immunodeficiency virus (HIV) exhibits extensive heterogeneity due to its rapid turnover, high mutation rate, and high frequency of recombination. Its remarkable genetic diversity plays a key role in virus adaptation, including development of drug resistance. The increasing complexity of antiretroviral regimens has favored selection of HIV variants harboring multiple drug resistance mutations. Evolution of drug resistance is characterized by severe fitness losses, which can be partially overcome by compensatory mutations or other adaptive changes that restore virus replication capacity. Recent reports have addressed the impact of drug-resistance mutations on viral fitness. Methods include in vitro estimates based on the determination of viral replication kinetics, viral infectivity in single-cycle assays and growth competition experiments; as well as estimates of the relative fitness of viral populations in vivo calculated from standard population genetics theory. This review focuses on the effects in viral fitness of mutations arising during treatment with reverse transcriptase and protease inhibitors, and the molecular mechanisms (including compensatory mutations) that improve the viral fitness of drug-resistant variants. PMID- 14754436 TI - Variety of interpretation systems for human immunodeficiency virus type 1 genotyping: confirmatory information or additional confusion? AB - The emergence of drug resistance remains a major problem during antiretroviral treatment of patients infected with human immunodeficiency virus type 1 (HIV-1). As phenotypic drug resistance is laborious and expensive to determine, and because numerous specific mutations are known to be correlated with different resistance patterns, genotyping of the reverse transcriptase and protease genes of HIV is fast becoming an integral part of HIV management in industrialized countries. A number of software-based interpretation systems have been developed for the interpretation of the resulting complex nucleotide sequences. These programs either employ rule-based algorithms or are based on a genotype-phenotype database. This paper reviews recent publications that compare different such systems, trying to identify the degree of discordance between different systems and the reasons underlying such discrepant interpretations. The highest discordance rate was observed for nucleoside reverse transcriptase inhibitors (NRTIs) followed by protease inhibitors (PIs) and non-nucleoside reverse transcriptase inhibitors (NNRTIs). For the NRTIs, it is the role of nucleoside analogue associated mutations, for the PIs and for the NNRTIs, that of secondary mutations that causes most discrepancies. As the complexity of the mutation pattern is likely to increase further with new drugs becoming available, rule based genotype interpretation algorithms need to be updated frequently. Whilst not recommending one particular system, the authors believe that the correlation of genotypic with clinical data is probably the best way to develop an optimal algorithm. PMID- 14754437 TI - Anti-HIV inhibitors based on nucleic acids: emergence of aptamers as potent antivirals. AB - The development of resistance and the inability of currently approved antiretroviral drugs to completely eradicate HIV-1 have led to increased focus on therapies other than small molecules. Although nucleic acid-based intervention requires complex tasks involving intracellular delivery and/or stable expression in target cells, recent advances in gene therapy methods combined with continued progress in stem cell approaches have made nucleic acid-based compounds excellent candidates for effectively inhibiting intracellular targets. Consequently, multiple nucleic acid-based therapies are being developed. These include antisense nucleic acids, peptide nucleic acids and RNA decoys, which can interfere with HIV-1 replication. More recently, RNA interference, which exploits a novel cellular pathway, has been shown to effectively reduce viral titers in cell culture and promises to be a potential candidate for suppressing HIV replication in vivo. A promising candidate in the midst of these emerging approaches is the aptamer approach, which involves the use of a class of small nucleic acid molecules isolated from combinatorial libraries by an in vitro evolution protocol termed SELEX. Aptamers exhibit exquisite specificity, high affinity and the virtual lack of immunogenicity, features that make them exceptionally well-suited to combat HIV without affecting the host. The powerful nature of these specific antagonists of protein function could lead to the development of an effective anti-HIV therapy. Several highly specific, nucleic acid aptamers targeting select HIV proteins have been described. Investigations with anti-HIV RNA aptamers have shown an effective block to viral replication. This review summarizes the existing nucleic-acid based approaches to block HIV replication and attempts to chart the current progress in the development of aptamers against HIV, their use in inhibiting the virus replication, prospects for their use in the clinic and potential drawbacks. PMID- 14754438 TI - Trace elements supplementation: recent advances and perspectives. AB - Supplementation of essential trace elements has become an increasingly important field of research in modern pharmacology. The present review presents general aspects related to the essentiality of inorganic systems in living organisms, followed by a detailed discussion of the supplementation of iron, copper, zinc, chromium, selenium, magnesium and some other minor trace elements. Some recent advances in this field as well as future challenges and perspectives are also discussed. PMID- 14754439 TI - Chemical and biological considerations in the treatment of metal intoxications by chelating agents. AB - Effective chelation treatment of metal intoxications requires that the pharmacokinetics of the administered chelator in fact leads to chelation of the toxic metal, preferably forming a less toxic species which is effectively excreted. This depends on physical and chemical characteristics of metals and chelators as e.g. ionic diameter, ring size and deformability, hardness/softness of electron donors and acceptors, administration route, bioavailability, metabolism, organ and intra/extra cellular compartmentalization, and excretion. In vivo chelation is unlikely to reach equilibrium determined by the standard stability constant, as rate effects and ligand exchange reactions as well as the pharmacokinetics of the chelator considerably influence complex formation. Hydrophilic chelators enhance renal metal excretion, but mainly their extracellular distribution limit their effect to mainly extracellular metal pools. Lipophilic chelators can decrease intracellular stores, but may redistribute toxic metals to e.g. the brain. In chronic metal induced disease, necessitating life-long chelation, toxicity and side effects of the chelator may limit the treatment. The metal selectivity of chelators is important, due to the risk of essential metals depletion. Dimercaptosuccinic acid and dimercaptopropionic sulfonate are presently gaining increased acceptance among clinicians, undoubtedly improving the management of human metal intoxications including lead, arsenic and mercury compounds. Still, development of new safer chelators suited for long-term oral administration for chelation of metal deposits, mainly iron, is an important challenge to the future research. PMID- 14754440 TI - Metal complexes as chemotherapeutic agents against tropical diseases: trypanosomiasis, malaria and leishmaniasis. AB - Parasitic diseases represent a major world health problem with very limited therapeutic options, most of the available treatments being decades old and suffering from limited efficacy and/or undesirable collateral effects. The use of metal complexes as chemotherapeutic agents against these ailments appears as a very attractive alternative. Although the design of metal complexes with good therapeutic index is still rather empirical, a number of potential metal-based antiparasitic drugs have become available. In this review, advances in the use of metal complexes for the treatment of trypanosomiasis, malaria, and leishmaniasis as important representatives of the general area of tropical diseases is described. PMID- 14754441 TI - The wide pharmacological versatility of semicarbazones, thiosemicarba-zones and their metal complexes. AB - The more significant bioactivities of a variety of semicarbazones (anti-protozoa, anticonvulsant) and thiosemicarbazones (antibacterial, antifungal, antitumoral, antiviral) and their metal complexes are reviewed together with proposed mechanisms of action and structure-activity relationships. Clinical or potential pharmacological applications of these versatile compounds are discussed. PMID- 14754442 TI - Pharmacokinetic study and trial for preparation of enteric-coated capsule containing insulinomimetic vanadyl compounds: implications for clinical use. AB - To treat patients suffering from diabetes mellitus, we developed several types of orally active vanadyl complexes to replace painful insulin injections, and prepared them in the form of enteric-coated capsules containing vanadium compounds. Pharmacokinetic analysis demonstrated that these capsules enhance the bioavailability of pharmacologically active vanadyl species. PMID- 14754443 TI - Antitumor titanium compounds. AB - Most anticancer titanium compounds act against tumors in the gastrointestinal tract. Activity towards breast, lung and skin (melanoma) cancers is shown by some as well. Among their appealing properties is that they do not show common side effects of widely used cytostatic agents such as emesis, alopecia or bone marrow impairment. These features make titanium compounds interesting for combined therapy and further study. This review focuses on two drugs that reached clinical trials, namely, titanocene dichloride and budotitane. We try to integrate the biological fate of the related Ti-cyclopentadienyl and Ti-beta-diketonato families of drugs, delineating the structure-activity relationship. We also discuss novel related species with increased solubility for improved drug delivery and some potentially useful polynuclear compounds. PMID- 14754444 TI - Rhenium-188 and copper-67 radiopharmaceuticals for the treatment of bladder cancer. AB - The favourable nuclear properties of copper-67 and rhenium-188 for therapeutic application are described, together with methods for the chemical synthesis of a number of derivatives. Survival from invasive bladder cancer has changed little over the past 20 years. The intravesicular administration of Cu-67 or Re-188 radiopharmaceuticals in the treatment of bladder cancer offers some promise for improvement in this situation. PMID- 14754445 TI - Beta-lactams in the new millennium. Part-I: monobactams and carbapenems. AB - Beta-lactam ring-containing compounds such as penicillins, ampicillin, amoxicillin, cephalosporins and carbapenem are among the most famous antibiotics. This article reviews the recent developments in the study of such compounds. The introductory paragraph, which highlights the significance of the subject and cites most of the leading references of the previous century, is followed by an overview of beta-lactams and some novel methodologies for the synthesis of bi-, tri- and polycyclic derivatives. The rest of the sections deal with design, synthesis and biological activity of monobactams and carbapenems. Many of them have potential antibacterial activity, even against some resistant strains, and enzyme inhibitory activity. PMID- 14754446 TI - Beta-lactams in the new millennium. Part-II: cephems, oxacephems, penams and sulbactam. AB - Beta-lactam ring-containing compounds such as penicillins, ampicillin, amoxicillin, cephalosporins and carbapenems are among the most famous antibiotics. This article reviews the recent developments in study of cephems, oxacephems, penams and sulbactam. Many of the compounds reviewed have potential antibacterial activity, even against resistant strains such as MRSA, and enzyme inhibitory activity. PMID- 14754447 TI - Nicotine and nicotinic receptor involvement in neuropsychiatric disorders. AB - Advances in the understanding of the neurobiology of the nicotinic receptor have started to be matched by an appreciation of the potential role of these receptors in a variety of neuropsychiatric disorders. While alterations in nicotinic receptor number and/or function have been associated with such conditions as Alzheimer's disease for several years, there is increasing evidence that nicotinic receptor function may play a significant role in other disorders as well including schizophrenia, Parkinson's disease, anxiety disorders, and attention deficit-hyperactivity disorder (ADHD). Research in our laboratory and those of other investigators have utilized sophisticated psychopharmacological, cognitive, electrophysiological, neuroimaging and other techniques to assess the impact of nicotinic receptor modulation on the clinical expression of these disorders. This manuscript reviews data, both experimental and clinical, relating to the role of nicotine and/or nicotinic receptor function in a variety of neuropsychiatric disorders with the perspective of developing appropriate targets for therapeutic drug development. PMID- 14754448 TI - Physiological roles of neuronal nicotinic receptor subtypes: new insights on the nicotinic modulation of neurotransmitter release, synaptic transmission and plasticity. AB - Nicotinic acetylcholine receptors (nAChRs) are widely expressed in the mammalian central nervous system (CNS). Despite this, very little was known, until recently, about their physiological role. In the periphery, nicotinic receptors mediate vital excitatory fast synaptic cholinergic transmission at both the neuromuscular junction and ganglia. In the brain, this role has been mainly "delegated" to glutamate receptors. The very broad cholinergic innervations of most brain areas, including the cortex, have implicated this system, and brain nicotinic receptors in particular, in a unique "modulatory" role of other transmitters systems. Recent evidence confirms, on one hand, that brain nicotinic receptors have a dominant "presynaptic" modulatory function, controlling the release of both acetylcholine (auto-receptors) and other neurotransmitters (hetero-receptors). On the other hand, more experimental data support the idea that a variable component of fast synaptic transmission in the brain can also be mediated by "postynaptic" nicotinic receptors, which, in turn, can control cell excitability. A challenging goal is to identify which one of the plethora of nicotinic receptor subtypes is mediating each effect in different brain areas, and which of these receptors and functions are lost or affected in different human neuro-psychiatric disorders. Needless to say, a better understanding of the physiological role of brain nicotinic receptors will drive our quest for more selective and efficacious nicotinic receptor targeted therapeutic agents. PMID- 14754449 TI - Design of ligands for the nicotinic acetylcholine receptors: the quest for selectivity. AB - In the last decade, nicotinic acetylcholine receptors (nAChRs) have emerged as important targets for drug discovery. The therapeutic potential of nicotinic agonists depends substantially on the ability to selectively activate certain receptor subtypes that mediate beneficial effects. The design of such compounds has proceeded in spite of a general shortage of data pertaining to subtype selectivity. Medicinal chemistry efforts have been guided principally by binding affinities to the alpha4beta2 and/or alpha7 subtypes, even though these are not predictive of agonist activity at either subtype. Nevertheless, a diverse family of nAChR ligands has been developed, and several analogs with promising therapeutic potential have now advanced to human clinical trials. This paper provides an overview of the structure-affinity relationships that continue to drive development of new nAChR ligands. PMID- 14754450 TI - Neuronal nicotinic acetylcholine receptor agonists: pharmacophores, evolutionary QSAR and 3D-QSAR models. AB - Neuronal nicotinic acetylcholine ion channel receptors (nAChRs) exist as several subtypes and are involved in a variety of functions and disorders of the central nervous system (CNS), such as Alzheimer's and Parkinson's diseases. The lack of reliable information on the 3D structure of nAChRs prompted us to focus efforts on pharmacophore and structure-affinity relationships (SAFIRs). The use of DISCO (DIStance COmparison) and Catalyst/HipHop led to the formulation of a pharmacophore that is made of three geometrically unrelated features: (i) an ammonium head involved in coulombic and/or H-bond interactions, (ii) a lone pair of a pyridine nitrogen or a carbonyl oxygen, as H-bond acceptor site, and (iii) a hydrophobic molecular region generally constituted by aliphatic cycles. The quantitative SAFIR (QSAFIR) study was carried out on about three hundred nicotinoid agonists, and coherent results were obtained from classical Hansch type approach, 3D QSAFIRs, based on Comparative Molecular Field Analysis (CoMFA), and trade-off models generated by Multi-objective Genetic QSAR (MoQSAR), a novel evolutionary software that makes use of Genetic Programming (GP) and multi objective optimization (MO). Within each congeneric series, Hansch-type equations revealed detrimental steric effects as the major factors modulating the receptor affinity, whereas CoMFA allowed us to merge progressively single-class models in a more global one, whose robustness was supported by crossvalidation, high prediction statistics and satisfactory predictions of the affinity data of a true external ligand set (r(2)(pred) = 0.796). Next, MoQSAR was used to analyze a data set of 58 highly active nicotinoids characterized by 56 descriptors, that are log P, MR and 54 low inter-correlated WHIM (Weighted Holistic Invariant Molecular) indices. Equivalent QSAFIR models, that represent different compromises between structural model complexity, fitting and internal model complexity, were found. Our attention was mostly engaged by a number of nonlinear QSAFIRs, which relate nAChR affinity with the log P and directional WHIM descriptors. The results reviewed herein show as QSAFIRs may helpfully complement the pharmacophores, thus enhancing the applicability of computer-aided methodologies in the field of nAChR agonists. PMID- 14754451 TI - Receptor binding thermodynamics at the neuronal nicotinic receptor. AB - Simple determination of K(A) or K(D) values makes it possible to calculate the standard free energy DeltaG degrees = -RTlnK(A) = RT lnK(D)(T= 298.15 K) of the binding equilibrium but not that of its two components as defined by the Gibbs equation DeltaG degrees = DeltaH degrees - TDeltaS degrees where DeltaH degrees and DeltaS degrees are the equilibrium standard enthalpy and entropy, respectively. Recently, it has been shown that the relative DeltaH degrees and DeltaS degrees magnitudes can often give a simple "in vitro" way for discriminating "the effect", that is the manner in which the drug interferes with the signal transduction pathways. This particular effect, called "thermodynamic discrimination", results from the fact that binding of antagonists may be enthalpy-driven and that of agonists entropy-driven, or vice-versa. In the past, the thermodynamic discrimination was reported for the beta-adrenergic G-protein coupled receptor (GPCR) and confirmed later for adenosine A(1), A(2A) and A(3) receptors. Moreover, it has been found that the binding of all ligand-gated ion channel receptors (LGICR) investigated was thermodynamically discriminated. In particular, affinity constants for typical neuronal nicotinic receptor ligands were obtained by both saturation and inhibition experiments with the radioligand [(3)H]-cytisine, a ganglionic nicotinic agonist. Thermodynamic parameters indicated that agonistic binding was both enthalpy- and entropy-driven, while antagonistic binding was totally entropy-driven. These results have shown that neuronal nicotinic receptor agonists and antagonists were thermodynamically discriminated. On these grounds, the thermodynamic behaviour makes it possible to discriminate drug pharmacological profiles in vivo through binding experiments in vitro. PMID- 14754452 TI - Nicotinic acetylcholine receptor agonists: a potential new class of analgesics. AB - Current analgesics, such as opioids and nonsteroidal anti-inflammatory drugs (NSAIDs), are largely refinements of approaches available for more than 100 years and have critical liabilities and limitations. A number of new molecular targets for analgesia have been proposed in recent years, including the neuronal nicotinic acetylcholine receptor (nAChR). Agonists at neuronal nAChRs have antinociceptive effects in a variety of preclinical pain models. Moreover, nicotine can decrease experimentally-induced pain in humans without disrupting normal tactile sensation. These data from both experimental animals and humans suggest that compounds targeting neuronal nAChRs may represent a new class of analgesic agents. In this paper, we provide brief overviews of the physiology of pain, the animal models used to assess potential analgesics preclinically, and the biology of nAChRs. We then provide a review of preclinical data on the antinociceptive effects of a variety of neuronal nAChR agonists and a discussion of potential mechanisms, including evidence that antinociception is mediated by activation of brainstem nuclei with descending inhibitory inputs to the spinal cord. An evaluation of the clinical potential of this approach must also consider potential side effects. Undesirable side effects of nicotine are well known, but as we will discuss in detail, these effects are not produced by all neuronal nAChR agonists and the existence of neuronal nAChR subtypes may provide a basis for separating therapeutic effects from toxicities. PMID- 14754453 TI - Actual targets in cytodifferentiation cancer therapy. AB - Transformation of a normal cell into a tumor cell results from six essential alterations in cell physiology. There is a complex relationship that exists between growth, differentiation, neoplastic transformation, and the expression of genes and tumor suppressor genes. The knowledge of these mechanisms demonstrates that it is possible to pharmacologically modulate the growth and differentiation of tumor cells. The differentiation therapy focuses on demonstrating that cancer is a reversible state with altered maturation in which the transformed phenotype may be suppressed by cytostatic agents and by the pharmacological differentiation towards benign forms with no proliferative potential. One of the mechanisms determining the activity of target genes is the post-translational modification of the N-terminal tails of core histones. Inappropriate repression of genes required for cell differentiation has been linked to several forms of cancer. Histone deacetylase inhibitors modulate transcription, and are endowed with cytodifferentiating, antiproliferative and apoptogenic properties. Retinoids modulate cell differentiation, proliferation, apoptosis and morphogenesis in vertebrates, and have proved to be clinically useful. Their biological effects are mediated by the activation of retinoic acid receptors, which are ligand dependent gene transcription factors. Checkpoints during cell cycle allow the cell to respond to proliferation signals or decide between the alternate pathways leading to cytokinesis, differentiation, quiescence, and cell death. Abrogation of normal cell cycle controls in tumor cells contributes to their inability to differentiate and the restoration of such controls in G1 can lead to the resumption of differentiation and terminal cell division. Chemical inhibitors of cyclin-dependent kinases have been reported to stimulate differentiation of tumor cell lines. PMID- 14754454 TI - 3D QSAR models of interactions between beta-tubulin and microtubule stabilizing antimitotic agents (MSAA): a survey on taxanes and epothilones. AB - In the last two decades, paclitaxel (Taxol), 1) has dominated the anticancer chemotherapy as one of the most important antimitotic agents. Despite its clinical success, it presents some limitations due to its low aqueous solubility or multidrug-resistance (MDR) susceptibility. Among new compounds sharing paclitaxel's mechanism of action, epothilones have emerged as very promising candidates and are currently under clinical trials. While the electron crystallography (EC) structure of tubulin with embedded paclitaxel is available, only hypotheses about epothilone binding upon the protein may be advanced. This review illustrates our efforts in the minireceptor modeling approach as the most recent advances in the field of three-dimensional quantitative structure-activity relationship (3D QSAR) studies involving taxanes, epothilones and the corresponding protein environment. PMID- 14754455 TI - Antitumor potential of aza-bioisosterism in anthracenedione-based drugs. AB - Aza-bioisosteres of anthracene-9,10-diones and of anthrapyrazoles comprise an innovative class of anticancer compounds. They are formally derived by introduction of one or more nitrogens into the carbocyclic ring system of the parent drugs. Bioisosteres exhibit extensive changes in the physico-chemical properties and in the interactions with the pharmacological targets, DNA and DNA topoisomerase II, when compared to the carbocyclic analogues. A favourable spectrum of activity, reduced side effects and a unique tropism for solid tumors make the new derivatives a very interesting family of drugs. In particular, a 2 aza-anthracene-9,10-dione and a 9-aza-anthrapyrazole derivative are presently undergoing advanced clinical trials and appear to be promising in view of their approval as anticancer drugs. PMID- 14754456 TI - Distamycin A as stem of DNA minor groove alkylating agents. AB - Analogues of naturally occurring antitumor agents, such as distamycin A, which bind in the minor groove of DNA, represent a new class of anticancer compounds currently under investigation. Distamycin A has driven researcher's attention not only for their biological activity, but also for its non intercalative binding to the minor groove of double-stranded B-DNA, where it forms strong reversible complex preferentially at the nucleotide sequences consisting of 4-5 adjacent AT base pairs. The pyrrole-amide skeleton of distamycin A has been also used as DNA sequence selective vehicles for the delivery of alkylating functions to DNA targets, leading to a sharp increase of its cytotoxicity, in comparison to that, very weak, of distamycin itself. In the last few years, several hybrid compounds, in which known antitumor derivatives or simple active moieties of known antitumor agents have been tethered to distamycin frames, have been designed, synthesized and tested. Several efforts have been made to modify DNA sequence selectivity and stability of the distamycin and the structural modifications have been based on replacement of pyrrole by other heterocycles and/or benzoheterocycles obtaining a novel class of minor groove binding molecules called lexitropsins. The role of the amidino moiety, by means of the substitution with various groups, which includes ionizable, acid or basic, and non-ionizable groups, has been also studied. The synthesis of a hybrid deriving among the combination of the distamycin A and naturally occurring alkylating agent has been also reported. Several classes of distamycin derivatives that have been reported in the published literature have been described in this review article. PMID- 14754457 TI - Recent studies on natural products as anticancer agents. AB - Cancer will be the major cause of death in the 21st century and natural products should provide novel and more effective anticancer agents. This review deals with new natural molecules liable to become anticancer drugs, as well as recent specific strategies for a selective treatment of cancer. The introduction presents the current state of the art on anticancer research. Beside, in the following subheadings we summarize our research on cytotoxic natural quinone methide-triperpenes and their analogues. We also discuss our results on the anti tumour promoting activity of natural naphthoquinones and their derivatives. PMID- 14754458 TI - A novel technique for quantifying changes in vascular density, endothelial cell proliferation and protein expression in response to modulators of angiogenesis using the chick chorioallantoic membrane (CAM) assay. AB - Reliable quantitative evaluation of molecular pathways is critical for both drug discovery and treatment monitoring. We have modified the CAM assay to quantitatively measure vascular density, endothelial proliferation, and changes in protein expression in response to anti-angiogenic and pro-angiogenic agents. This improved CAM assay can correlate changes in vascular density with changes seen on a molecular level. We expect that these described modifications will result in a single in vivo assay system, which will improve the ability to investigate molecular mechanisms underlying the angiogenic response. PMID- 14754459 TI - Evidence for existence of thirty hypothetical proteins in rat brain. AB - BACKGROUND: The rapid completion of genome sequences has created an infrastructure of biological information and provided essential information to link genes to gene products, proteins, the building blocks for cellular functions. In addition, genome/cDNA sequences make it possible to predict proteins for which there is no experimental evidence. Clues for function of hypothetical proteins are provided by sequence similarity with proteins of known function in model organisms. RESULTS: We constructed a two-dimensional protein map and searched for expression of hypothetical proteins in rat brain. Two dimensional electrophoresis (2-DE) with subsequent in-gel digestion of spots and matrix-assisted laser desorption/ionization (MALDI) spectrometric identification were applied. In total about 3700 spots were analysed, which resulted in the identification of about 1700 polypeptides, that were the products of 190 different genes. A number of hypothetical gene products were detected (30 of 190, 15.8%) and are considered brain proteins. CONCLUSIONS: A major finding of this study is the demonstration of the existence of putative proteins that were so far only deduced from their nucleic acid structure by a protein chemical method independent of antibody availability and specificity and unambiguously identifying proteins. PMID- 14754460 TI - Biological effects of power frequency magnetic fields: Neurochemical and toxicological changes in developing chick embryos. AB - BACKGROUND: There are several reports that indicate a linkage between exposure to power frequency (50 - 60 Hz) magnetic fields with abnormalities in the early embryonic development of the chicken. The present study was designed to understand whether power frequency electromagnetic fields could act as an environmental insult and invoke any neurochemical or toxicological changes in developing chick embryo model. METHODS: Fertilized chicken eggs were subjected to continuous exposure to magnetic fields (50 Hz) of varying intensities (5, 50 or 100 microT) for a period of up to 15 days. The embryos were taken out of the eggs on day 5, day 10 and day 15. Neurochemical (norepinephrine and 5 hydroxytryptamine) and amino acid (tyrosine, glutamine and tryptophan) contents were measured, along with an assay of the enzyme glutamine synthetase in the brain. Preliminary toxicological investigations were carried out based on aminotransferases (AST and ALT) and lactate dehydrogenase activities in the whole embryo as well as in the liver. RESULTS: The study revealed that there was a significant increase (p < 0.01 and p < 0.001) in the level of norepinephrine accompanied by a significant decrease (p < 0.01 and p < 0.001) in the tyrosine content in the brain on day 15 following exposure to 5, 50 and 100 microT magnetic fields. There was a significant increase (p < 0.001) in glutamine synthetase activity resulting in the significantly enhanced (p < 0.001) level of glutamine in the brain on day 15 (for 100 microT only). The possible mechanisms for these alterations are discussed. Further, magnetic fields had no effect on the levels of tryptophan and 5-hydroxytryptamine in the brain. Similarly, there was no effect on the activity of either aminotransferases or lactate dehydrogenase in the whole embryo or liver due to magnetic field exposure. CONCLUSIONS: Based on these studies we conclude that magnetic field-induced changes in norepinephrine levels might help explain alterations in the circadian rhythm, observed during magnetic field stress. Also, the enhanced level of glutamine can act as a contributing factor for developmental abnormalities. PMID- 14754461 TI - Arachidonoyl-phospholipid remodeling in proliferating murine T cells. AB - BACKGROUND: Previous studies have shown that the functional capacity of T cells may be modulated by the composition of fatty acids within, and the release of fatty acids from membrane phospholipids, particularly containing arachidonic acid (AA). The remodeling of AA within membrane phospholipids of resting and proliferating CD4+ and CD8+ T cells is examined in this study. RESULTS: Splenic T cells were cultured in the presence or absence of anti-CD3 mAb for 48 h then labeled with [3H]AA for 20 min. In unstimulated cells, labeled AA was preferentially incorporated into the phosphoglycerides, phosphatidylcholine (PC) followed by phosphatidylinositol (PI) and phosphatidylethanolamine (PE). During a subsequent chase in unlabeled medium unstimulated CD4+ and CD8+ T cells demonstrated a significant and highly selective transfer of free, labeled AA into the PC pool. In contrast, proliferating CD4+ and CD8+ T cells distributed labeled [3H]AA predominantly into PI followed by PC and PE. Following a chase in AA-free medium, a decline in the content of [3H]AA-PC was observed in association with a comparable increase in [3H]AA-PE. Subsequent studies revealed that the cold AA content of all PE species was increased in proliferating T cells compared with that in non-cycling cells, but that enrichment in AA was observed only in the ether lipid fractions. Finally, proliferating T cells preincubated with [3H]AA exhibited a significant loss of labeled arachidonate in the PC fraction and an equivalent gain in labeled AA in 1-alk-1'-enyl-2-arachidonoyl-PE during a chase in unlabeled medium. CONCLUSION: This apparent unidirectional transfer of AA from PC to ether-containing PE suggests the existence of a CoA-independent transacylase system in T cells and supports the hypothesis that arachidonoyl phospholipid remodeling may play a role in the regulation of cellular proliferation. PMID- 14754462 TI - Do recent data from the Seychelles Islands alter the conclusions of the NRC Report on the toxicological effects of methylmercury? AB - In 2000, the National Research Council (NRC), an arm of the National Academy of Sciences, released a report entitled, "Toxicological Effects of Methylmercury." The overall conclusion of that report was that, at levels of exposure in some fish- and marine mammal-consuming communities (including those in the Faroe Islands and New Zealand), subtle but significant adverse effects on neuropsychological development were occurring as a result of in utero exposure. Since the release of that report, there has been continuing discussion of the public health relevance of current levels of exposure to Methylmercury. Much of this discussion has been linked to the release of the most recent longitudinal update of the Seychelles Island study. It has recently been posited that these findings supercede those of the NRC committee, and that based on the Seychelles findings, there is little or no risk of adverse neurodevelopmental effects at current levels of exposure. In this commentary, members of the NRC committee address the conclusions from the NRC report in light of the recent Seychelles data. We conclude that no evidence has emerged since the publication of the NRC report that alters the findings of that report. PMID- 14754463 TI - Willingness to pay for prevention and treatment of lymphatic filariasis in Leogane, Haiti. AB - BACKGROUND: Lymphatic filariasis (LF) affects more than 120 million people worldwide. Efforts to eliminate this disease require sustained community participation. This study explores community valuation of LF elimination efforts by estimating household and community willingness to pay (WTP) for the prevention of transmission and treatment of filarial lymphedema in the community of Leogane, Haiti. METHODS: A contingent valuation survey was used to assess individual WTP for specific prevention and treatment interventions. A 2-dimensional Monte Carlo simulation was developed to estimate confidence limits in mean WTP and to generate a distribution of WTP for the community, accounting for uncertainty in regression coefficients and variability within the population. RESULTS: Mean WTP was estimated to be $5.57/month/household (95% CL: $4.76, $6.72) to prevent disease transmission, and $491/yr (95% CL: $377, $662) for treatment of lymphedema for one person. Based on the estimated distributions, 7% and 39% of households were not willing to pay for prevention and treatment, respectively. CONCLUSIONS: These results suggest that the majority of the community places a positive value on both prevention and treatment of LF. Mean WTP provides a useful monetary estimate of overall societal benefit of LF prevention and treatment programs. However, for interventions which require broad and sustained community participation, the lower end of the distribution of WTP has additional implications. Cost recovery policies may result in inadequate participation and longer program duration. PMID- 14754464 TI - Materializing research promises: opportunities, priorities and conflicts in translational medicine. AB - There is considerable evidence that the translation rate of major basic science promises to clinical applications has been inefficient and disappointing. The deficiencies of translational science have often been proposed as an explanation for this failure. An alternative explanation is that until recently basic science advances have made oversimplified assumptions that have not matched the true etiological complexity of most common diseases; while clinical science has suffered from poor research practices, overt biases and conflicts of interest. The advent of molecular medicine and the recasting of clinical science along the principles of evidence-based medicine provide a better environment where translational research may now materialize its goals. At the same time, priority issues need to be addressed in order to exploit the new opportunities. Translational research should focus on diseases with global impact, if true progress is to be made against human suffering. The health outcomes of interest for translational efforts need to be carefully defined and a balance must be struck between the subjective needs of healthcare consumers and objective health outcomes. Development of more simple, practical and safer interventions may be as important a target for translational research as the development of cures for diseases where no effective interventions are available at all. Moreover, while the role of the industry is catalytic in translating research advances to licensed interventions, academic independence needs to be sustained and strengthened at a global level. Conflicts of interest may stifle translational research efforts internationally. The profit motive is unlikely to be sufficient alone to advance biomedical research towards genuine progress. PMID- 14754465 TI - New perspectives with antimetabolites in the management of breast cancer. PMID- 14754466 TI - Single-agent gemcitabine in the treatment of advanced breast cancer. AB - Despite the availability of a number of active cytotoxic agents and combination regimens derived from them, the median survival of patients with metastatic breast cancer (MBC) has not been dramatically prolonged and it remains largely an incurable disease. As such, common goals of treatment in this setting are to ameliorate cancer-related symptoms and maintain or improve patient quality of life for as long as possible. Achieving optimal palliation, however, requires balancing the toxicities of therapy with the cancer-controlling benefits, and, to this end, the development of effective and well-tolerated regimens is a priority. Gemcitabine, a novel nucleoside analogue with demonstrated antitumor activity and a favorable safety profile, has been evaluated in a number of recent clinical trials as single-agent therapy for MBC, including studies of first- and second line therapy, as well as the salvage setting for patients with taxane- and anthracycline-refractory advanced disease. PMID- 14754467 TI - Gemcitabine and taxanes as a new standard of care in breast cancer. AB - Advances in breast cancer continue to focus on the development of novel agents as well as the development of novel combinations, with the major therapeutic goal of treatment strategies revolving around improving response rates, delaying the time to disease progression, palliating symptoms, and improving quality of life. The taxanes are recognized as some of the most active single agents in breast cancer and demonstrate remarkable activity with manageable toxicity in combination with gemcitabine. Gemcitabine, a novel S-phase specific cytidine nucleoside analogue of deoxycytidine, has broad antitumor activity with significant monotherapy activity in breast cancer, with response rates ranging from 22% to 42% depending on the pretreatment characteristics of the patients. In general, gemcitabine s favorable single-agent activity and novel mechanism of action, in addition to its largely nonoverlapping toxicities, have facilitated its further development in combination with a variety of chemotherapy agents, including the taxanes. Several phase I and II trials have reported impressive activity for the gemcitabine/taxane doublet with the suggestion of clinical synergism between these 2 classes of agents. Given the remarkable and durable activity reported for this doublet, subsequent phase II trials have focused on optimizing doses and schedules. Unmistakably, these trial results are clear improvements over the single-agent activity of the taxanes in metastatic breast cancer, with the recently reported phase III trial comparing gemcitabine plus paclitaxel versus paclitaxel alone clearly reinforcing the superior outcomes demonstrated for the combination. Either as a single agent or in combination with the taxanes, gemcitabine remains a rational choice for the treatment of breast cancer. PMID- 14754468 TI - Gemcitabine and platinum combinations in patients with breast cancer previously treated with anthracyclines and/or taxanes. AB - As anthracycline-based regimens have become a standard treatment and are frequently used in the adjuvant therapy of breast cancer, the number of anthracycline-resistant cancers has begun to increase. Taxanes have also become more commonly used in the first-line metastatic and adjuvant setting, producing a need for new treatment options that are not cross-resistant with anthracyclines or taxanes and that have a relative non-overlapping toxicity profile with these agents. The combination of gemcitabine/cisplatin has been shown to have synergistic cytotoxic activity in vitro in breast cancer cell lines. In addition, several phase II trials have suggested that this combination is feasible and active in patients who have received prior anthracycline and/or taxane therapy. PMID- 14754469 TI - Gemcitabine and trastuzumab combinations for patients with metastatic breast cancer overexpressing HER2/neu. AB - In HER2/neu-positive metastatic breast cancer, the combination of chemotherapy and trastuzumab has become a standard of care. This review discusses HER2/neu overexpression in breast cancer and the use of trastuzumab-based therapies. Specifically, the rationale for a gemcitabine/trastuzumab combination in this disease entity and the available clinical data on the use of the combination are discussed. Response rates of 12%-42% have been seen with single-agent gemcitabine and 37%-62% with trastuzumab/gemcitabine combinations. Further work is currently ongoing to examine this promising combination. PMID- 14754470 TI - Rationale and clinical trial design for evaluating gemcitabine as neoadjuvant and adjuvant therapy for breast cancer. AB - Over the past 30 years, much progress has been made in adjuvant chemotherapy for patients with early-stage breast cancer. The introduction of anthracyclines and, recently, taxanes has significantly improved the efficacy of adjuvant chemotherapy and has provided useful insight regarding future development of even more efficacious regimens. However, despite significant progress, there remains considerable room for improvement. Although the manipulation of chemotherapy doses has not been found to be a successful strategy, manipulation of the interval of chemotherapy administration recently showed promise. It appears, nonetheless, that introducing new chemotherapy agents into the adjuvant setting is a more promising strategy than attempting to optimize the dose and schedule of existing agents. Gemcitabine has demonstrated significant antitumor activity in patients with advanced breast cancer when used either as a single agent or in combination with other active drugs (as a doublet or a triplet). Most of the available information to date has been obtained by combining gemcitabine with anthracyclines and/or taxanes. This approach has shown considerable antitumor activity and a reasonable toxicity profile, making it an important strategy for developing new adjuvant and neoadjuvant chemotherapy regimens. Several currently ongoing and planned adjuvant trials are incorporating gemcitabine (either sequentially or in combination) into anthracycline/taxane regimens. These trials will be important in establishing a role for this agent in the adjuvant setting and will hopefully lead to the development of more effective adjuvant chemotherapy regimens in the future. PMID- 14754472 TI - Two sessions with Catherine. PMID- 14754475 TI - Homosexuality: coming out of the confusion. AB - The author asserts that earlier theoretical assumptions contribute to the conceptual confusion about homosexuality in much contemporary psychoanalytic research. Bergeret's article, recently published in this Journal, is exemplary of this confusion. The author refutes his contention that homosexuality is not 'true' sexuality but merely a defensive, narcissistic fixation away from, or a nearly psychotic denial of, heterosexuality. He then clarifies specific areas of conceptual confusion regarding homosexuality still prevalent in psychoanalytic discourse that derive from earlier theoretical premises. These areas of confusion include manifest versus latent homosexuality, narcissism and bisexuality, oedipal dynamics and development, and transference and technique. PMID- 14754476 TI - Psychic bisexuality and creativity. AB - This contribution is complementary to a previous publication (Ferraro, 2001), which examined the role of bisexuality in psychopathology. This second article concentrates on the relationship between psychic bisexuality and creativity. After a brief clarification regarding the relationship between psychic bisexuality and option of gender, the author takes up two meanings of the bisexuality concept, both of which are of pre-eminent significance to him. The first is psychic bisexuality as a quality of the self related to the feminine and masculine as pure elements; the second is psychic bisexuality as an expression of identification with both parents, mother and father. The author presents the thoughts of various authors who have examined the link between psychic bisexuality and creativity, based on the same foundation, and then puts forward the hypothesis that in some blocks of creativity an alteration to psychic bisexuality can be traced. This hypothesis is illustrated through two clinical cases that focus on the dynamics impeding creative capacities and illustrate how these dynamics are gradually overcome. In the first more detailed case, he presents a lack of masculine elements, while in the second, using a brief part of an analysis, he presents a predominant lack of feminine elements. PMID- 14754477 TI - Words that touch. AB - In this paper the author examines her own use of language as a psychoanalyst and asks: what is the best way to help analyse and to find the words to express not only what they are thinking but also what they are feeling and experiencing? In common with other psychoanalysts, the author has observed that each of us simultaneously utilises both advanced psychic mechanisms that are accessible to symbolism and more archaic ones, which are less so. However, she draws a distinction between people who are able to tolerate the perception of their own heterogeneity, even if it is sometimes a source of suffering, and those whom she terms 'heterogeneous patients'. Patients in the latter category, whose lack of internal cohesion causes them anxiety, are afraid of losing their sense of identity. The author asks how we can understand their language and how we should speak to them. She uses several clinical examples to demonstrate that 'heterogeneous patients' need to be touched with a language that does not confine itself to imparting thoughts verbally but also conveys feelings and the sensations that accompany those feelings. It is also an 'incarnated' language because the words pronounced by the analyst can awaken, or reawaken, bodily fantasies in the patient. These words may enable him to find an emotional meaning in forgotten sensory or bodily experiences, which may then become a starting point for his work of thinking and of symbolisation. PMID- 14754478 TI - The patient's material as an aid to the disciplined working through of the countertransference and supervision. AB - The author argues that the patient's largely unconscious observations of the analyst's functioning are, at times, communicated in the patient's material and that this can impart a sense of clinical relevance to the countertransference. The concept of 'understanding work' is used to provide a psychoanalytical model of this phenomenon. This is illustrated in a clinical case and it is argued that a selective consideration of the patient's material can provide a proper discipline which steers the analyst between the twin dangers of megalomania, on the one hand, and involvement in a symmetrical, self-disclosing relationship, on the other. The author then applies these ideas to supervision and uses them to distinguish psychoanalytic supervision from a practice that also derives from an intersubjective paradigm but which, to the author's mind, is not distinct enough from personal analysis. PMID- 14754479 TI - Time and the apres-coup. AB - The author explores the different temporalities of developmental time, apres-coup and what she calls 'reverberation time'. She considers the paradoxical temporality of the 'here and now', showing that it is not pure present, and describes at the micro-level of sessional material how progressive and retrospective time go inherently together, one being a requisite for the other. PMID- 14754480 TI - Imre Hermann's Freudian theory of attachment. AB - In this paper the author addresses some problems concerning the relation between attachment theory and psychoanalysis and sketches the outline of a Freudian theory of attachment, based on the ideas of the Hungarian psychoanalyst Imre Hermann. First, he elucidates the fundamental divergence behind the insults and misunderstandings that have dominated the debate between attachment theory and Freudian psychoanalysis: they differ radically in their conceptions of temporality and of the relation between psychopathology and human nature. Second, the author presents Hermann's work on 'Clinging--Going-in-search' (1976) as a theoretical model in which the findings of attachment theory can be integrated in a theory of psychopathology that is radically dimensional and that preserves the methodology of Freudian psychoanalysis. In the third part of the paper, the author discusses the question of whether Hermann's 'clinging instinct' is a primary instinct in the Freudian sense, and whether it is an interesting alternative for Freud's 'death instinct', as Hermann claims. PMID- 14754481 TI - Knowledge and experience of time in primitive mental states. AB - The author proposes that time be considered an object of working through during the psychoanalysis of patients whose mental functioning reveals distinct primitive aspects. These so-called 'difficult patients' are particularly intolerant of the temporal limits of analytic sessions and often attempt to undermine the analytic setting. He presents some hypotheses about time and the mind's depth levels. A series of clinical vignettes taken from the analysis of adolescent, borderline and psychotic patients shows several ways in which the analysand's perception of and relation to time surfaces and is worked through in the course of analysis. Analyzing the relationship with time and the conflict between denial and acceptance of temporality proved a significant catalyst in the development of cases that had tenaciously impeded progress. It thus became possible to bring about important changes in the ways such patients experienced affect in the analytic relationship and in object relations, not by acting directly upon these relationships but by working instead on the formal parameters that organize the mind-and primarily on its spatio-temporal organization. PMID- 14754482 TI - A Buddhist contribution to the psychoanalytic psychology of self. AB - The author attempts to integrate the concepts of self used in psychoanalytic theory with the understanding of the nature of self as explained within the Buddhist meditative tradition. He divides different concepts of self in psychoanalytic theory into three major levels of consciousness and abstraction: self as experience, representational self and self as system. The representational level is defined as consisting of unconscious organizing structures of interaction: the system level is a hierarchically higher organization of representations, while the experiential level consists of the moment-to-moment flow of consciousness. He argues that for the sake of theoretical clarity these levels should be differentiated in discussions of self. He then describes the Buddhist psychology of self and tries to show how this perspective can enrich psychoanalytic understanding of the experiential self and of narcissism, which in Buddhist language would be described as clinging to (seeking or avoiding) images of self that arise in the mind. Last, he describes a model of therapeutic development using different levels of self and the interrelationship between them, showing how psychoanalytic psychotherapy and Buddhist insight meditation emphasize different levels of self using complementary rather than mutually exclusive methods. PMID- 14754483 TI - Transition to young adulthood: a prospective study. AB - In this paper, the authors report on the transition to young adulthood (18-25) in the highest functioning 18 individuals in our cohort (GAF > or = 90) from a prospective longitudinal study of 76 lives followed from birth. These 18 individuals provide as clear a view as possible into the inner lives of people least distorted by psychopathology. This gives us a more crystalline photograph of psychic structure: the precipitant of family, society and psyche itself. Using a standard DSM IV criterion, Global Assessment Functioning (GAF), provides phenomenological homogeneity, but leaves us with heterogeneity of inner life. The 18 individuals had relatively rare drug or alcohol use and less sexual partners. Vocational functioning exceeded intimacy. Four had tumultuous pathways into adulthood. Nine were crossover cases, coming from sub-optimal mothering, with significant variation in gender distribution and intrapsychic structure. The paper discusses factors facilitating and inhibiting development and the nature of memories. PMID- 14754484 TI - Erna and Melanie Klein. AB - Erna was one of the child patients treated by Melanie Klein in Berlin, employing her recently discovered play technique. Since Erna died in Chile, the authors considered the IPA Congress in Santiago an opportunity to present a paper as a homage both to Erna and, especially, to Klein. She learned much from that very disturbed child, which she later used to sustain the ongoing development of her theories. The paper explores biographic data relevant to understanding both the case and the theories. It analyses the case material to follow Klein in the discovery and the handling of the child's transference and the harsh expressions of hate, jealousy and envy, which are brought in, with sad consequences, by strong persecutory feelings. Klein's comparison of this case with that of Freud's Wolf-man is also considered, mostly to show that the similarities were less than originally claimed, and that Klein, perhaps, was introducing a theoretic shift which led her technique to gradually change from 'Nachtraglichkeit' to the 'signification-resignification' pair, akin to Strachey's concept of the mutative interpretation. Lastly, the comprehension of Erna's strong psychotic traits and the links with later developments of the theory on psychosis are studied. PMID- 14754485 TI - The capacity to be an analyst: A contribution from attachment research to the study of candidate selection. AB - In this paper the author discusses how the study of candidate selection, once a topic of vibrant research, has unfortunately languished. Certain qualities were thought to characterize the successful candidate. However, they were never successfully operationalized nor empirically tested. Possibly because of this lack of empirical data, selectors today have difficulty articulating their criteria and are relying on intuition. In order to provide a more rational basis for contemporary selection, the author looks to the attachment literature. This makes sense because attachment theory shares some basic assumptions of contemporary psychoanalysis. The Adult Attachment Interview (AAI) is a research tool that predicts the ability of a parent to convey attachment security. It is scored by attending to how a person speaks about his early attachment experiences. The AAI appears to tap into similar qualities to those selection researchers have sought in their candidates. Further, the scoring method of the AAI appears to be similar to the last attempt by selection researchers to operationalize them. Given these similarities, the author recommends an empirical study using the AAI to operationalize these qualities in analytic candidates. The study would test their importance for success in the training program, thus offering selectors some empirical grounding for their choices. PMID- 14754489 TI - Chloroquine efficacy in the treatment of uncomplicated malaria at three sentinel sites in northern Togo. AB - In Togo, chloroquine (CQ) remains the first-line drug for the treatment of uncomplicated, Plasmodium falciparum malaria. In the absence of recent data on the level of parasite resistance to antimalarial drugs, Togo's National Malaria Control Programme (NMCP) decided to assess the current efficacy of CQ in the treatment of uncomplicated, P. falciparum malaria at three sentinel sites in the north of the country. Between the September and November of 2001, the World Health Organization's standard 14-day protocol was used to investigate 153 malarious children aged 6-59 months old (46 from Sokode, 54 from Niamtougou and 53 from Dapaong). Of the subjects from Sokode, Niamtougou and Dapaong, early treatment failure was observed in 0%, 7% and 12%, late treatment failure in 0%, 11% and 17%, and overall parasitological failure in 0%, 45% [with a 95% confidence interval (CI) of 39%-51%] and 62% (CI=54%-70%), respectively. Even within northern Togo, there is clearly considerable geographical variation in the level of resistance to CQ. Before an efficient antimalarial-drug policy can be developed, there is an urgent need to develop and use the national surveillance system further, to collect relevant data on the efficacies of CQ and other antimalarial drugs, such as amodiaquine and sulfadoxine-pyrimethamine. PMID- 14754491 TI - The burden of malaria in Ahmedabad city, India: a retrospective analysis of reported cases and deaths. AB - Owing to the paucity of accurate information on the burden of malaria in urban India, a retrospective, epidemiological study was carried out in Ahmedabad city, which has a population of about 3 million. Surveillance data for the years 1965 1998 showed a gradual resurgence of malaria between 1967 to 1976, followed by waves of low and high incidences. Plasmodium vivax always predominated but the proportion of cases attributed to P. falciparum increased markedly from 1983. When the surveillance data and health records of the major public and private health facilities in the city were analysed, for the period between 1991 and 1998, P. vivax was found to account for 69% of all malaria cases and P. falciparum for the other 31%. The incidence of infection with each Plasmodium species showed seasonal variation, with that of P. vivax increasing from January to September but then declining as the incidence of P. falciparum increased. The age-specific differences seen in incidence were not statistically significant (P=0.7). The annual numbers of malaria-attributable deaths were strongly correlated with the incidence of P. falciparum (r=0.88). The malaria incidence detected (37431 cases, representing a mean annual incidence of 12.2 cases/1000) was nine times greater than that officially reported (4119 cases, or 1.3 cases/1000 each year). Similarly, the annual malaria-attributable mortality detected (22 deaths/million) was far higher than that officially notified (0.3 death/million). The results of the retrospective analysis not only provide a more accurate, baseline estimate of the burden of malaria in an urban area of India but also clearly indicate the need for a much more efficient health-information system, for recording and managing malaria in such a setting. PMID- 14754490 TI - Therapeutic efficacies of antimalarial drugs in the treatment of uncomplicated, Plasmodium falciparum malaria in Assam, north-eastern India. AB - In the Indian state of Assam, the current therapeutic efficacies of the drugs commonly used in the area for the treatment of uncomplicated, Plasmodium falciparum malaria were investigated. As is routine in this area, subjects found positive for P. falciparum malaria were initially treated with chloroquine (CQ). They were given sulfadoxine-pyrimethamine (SP) if this treatment failed, and subsequently quinine if the SP failed. The protocol of the World Health Organization's extended in-vivo test was used to follow parasite clearance and clinical cure. Therapeutic response was assessed by comparing the baseline (day 0) level of parasitaemia with that observed on day 3. Many (75.7%) of the 144 evaluable subjects were treatment successes after CQ, but six early (4.2%) and 29 (20.1%) late CQ-treatment failures were observed. Of the 34 CQ-treatment failures followed, 31 (91.2%) responded adequately to SP but the other three were early (one) or late (two) SP-treatment failures. Two (66.7%) of the SP-treatment failures responded adequately to parenteral quinine but the other (a late quinine treatment failure) had to be given an artemisinin derivative to achieve a clinical cure. The foci in which multidrug-resistant cases of malaria are developing in India need to be identified quickly, so that such cases can be cured before the mutant strains of P. falciparum that are resistant to several drugs have a chance to become more widespread. PMID- 14754492 TI - Mast cells, histamine and the pathogenesis of intestinal damage in experimental Trypanosoma brucei brucei infections. AB - Intestinal damage with increased permeability is a prominent feature of experimental African trypanosomiasis. The possible involvement of mast cells and histamine in the altered gut integrity was investigated, at the level of the jejunum, in BALB/c mice infected with Trypanosoma brucei brucei. Mast cells were studied by selective staining of granule content with Alcian Blue/Safranin and quantitative histology, and histamine concentrations were determined by a fluorimetric method. Mast-cell activation, shown by a marked reduction in the numbers of positive-staining cells seen per villous section, was prominent on days 7 and 14 post-infection (there was, for example, a reduction to 36% of the control value by day 14; P=0.0001). By day 21, however, there were 131% more staining cells per villous section in the infected mice than in the uninfected controls (P=0.003). Histamine levels in homogenates of the jejunal mucosae of the infected mice were found to be significantly elevated at each time-point. The maximum increase was observed on day 14, when the numbers of granulated mast cells were at their lowest, with mean (S.E.) concentrations of 6.744 (0.890) ng/mg tissue for the infected mice and 2.813 (0.321) ng/mg for the uninfected controls (P=0.0008). The jejunal mucosa suffered progressive morphological damage during the infection, with oedema of the lamina propria and villi and disruption of the endothelium. These results indicate that mast cells are involved with the intestinal pathology that develops during experimental African trypanosomiasis. PMID- 14754493 TI - The PCR-based detection of Leishmania major infections in Meriones libycus (Rodentia: Muridae) from southern Iran. AB - Cutaneous leishmaniasis (CL) is an increasing public-health problem in the countries of the Mediterranean region. In southern Iran, for example, the incidence of CL has more than doubled over the last decade. As part of an investigation of this worrying trend, the leishmanial infections of the wild rodents that may act as reservoir hosts were investigated in the Iranian province of Fars. Overall, 82 rodents, 56 of them Libyan jirds (Meriones libycus), were collected, in live traps, from a new, hypo-endemic focus of zoonotic CL. When Giemsa-stained smears of ear tissue or any lesions on the rodents were prepared and checked under a light microscope, amastigotes were found in the smears from six of the rodents, all M. libycus. None of the other rodents investigated was found to be infected. The infected rodents were encountered in two of the three areas studied. In PCR-based assays, the amastigotes seen in the ear smears and those from two recent human cases of CL from the focus were all found to be identical to a reference strain of L. major. It therefore seems that M. libycus is the main active reservoir host for ZCL in the focus. PMID- 14754494 TI - The antibody responses to adult-worm antigens of Schistosoma haematobium, among infected and resistant individuals from an endemic community in southern Ghana. AB - Antibody responses to antigens from adult Schistosoma haematobium were investigated in an endemic community in Ghana, using microplate-ELISA. The results of a survey of egg output in urine and of a questionnaire-based investigation of water-contact activities were used to select 'endemic normal' (EN) and patently infected (PI) individuals as subjects. The plasma levels of antibodies reacting with the adult-worm antigens were determined and compared and the correlations between these levels and the age, water-contact index and egg output of each subject were evaluated. Compared with the EN subjects, the PI generally had higher levels of anti-worm IgG and IgE but lower levels of anti worm IgA. When the data for the EN and PI groups were combined, the levels of anti-worm IgG and IgE were found to be positively correlated with egg output and with each other. Whichever the antibody class considered, levels of anti-worm antibodies were never negatively correlated with egg output. These results indicate that anti-worm IgE and IgG could be used as markers to reflect current infection intensity, and that anti-worm antibodies may not act as protective antibodies in the natural course of urinary schistosomiasis. PMID- 14754495 TI - Impact of long-term ivermectin (Mectizan) on Wuchereria bancrofti and Mansonella perstans infections in Burkina Faso: strategic and policy implications. AB - Parasitological and clinical surveys were used to determine the long-term impact of ivermectin on the prevalence of Wuchereria bancrofti and Mansonella perstans filarial infections, when the drug was given under community-directed-treatment strategies for onchocerciasis control. The study was undertaken in 11 communities in south-western Burkina Faso. Six of the villages investigated had been treated with ivermectin at least once a year for five of 6 years, with a mean coverage of approximately 65% in each round. The other five, adjacent villages, which were matched with the ivermectin-treated communities by size, ethnicity and social and economic activities, had never been treated because they were not endemic for onchocerciasis. Each subject was checked by the microscopical examination of a smear of 'night' blood, by measurement of the level of circulating antigens from adult W. bancrofti, and by clinical examination for hydrocele (if male) and lymphoedema. The prevalences of lymphoedema and hydrocele in the treated villages were similar to those in the untreated. The prevalences and intensities of W. bancrofti and M. perstans microfilaraemia were, however, significantly lower in the ivermectin-treated communities. The implications of this study are discussed in relation to the old Onchocerciasis Control Programme (OCP) and to the ongoing African Programme for Onchocerciasis (APOC), where extensive and sustained ivermectin distribution is planned through community-based treatment programmes. As with onchocerciasis in Africa, the success of annual treatments to control lymphatic filariasis will depend not only on the number of regular rounds of treatment given but on adequate coverages being achieved in each round. Wherever ivermectin is being distributed alone, for onchocerciasis control, its impact on other filarial infections, notably W. bancrofti, should be evaluated routinely. Any opportunity to add donated albendazole to such distributions should be taken, both to limit the transmission of W. bancrofti and for the wider public-health benefits. PMID- 14754496 TI - A qualitative study of the perceptions, practices and socio-psychological suffering related to chronic brugian filariasis in Kerala, southern India. AB - Lymphatic filariasis is a major health problem in many parts of the tropical world. Although the disease itself is rarely fatal, the disability caused by the swollen extremities, the acute attacks of adenolymphangitis and the consequent sufferings of those afflicted are considerable. The economic burden imposed by lymphatic filariasis is not fully quantified and information on the social and psychological problems caused by the disease is scanty. Semi-structured interviews were therefore used, in southern India, to assess the perceptions, practices and socio-psychological problems of 127 patients with brugian filariasis. The patients were aware of the causative factors and the precautions to be taken to prevent progression of the disease. However, depression and loss of job opportunities were common in the study population. Patients also complained that the disease eroded their standing in the community and diminished their prospects of marriage. Awareness of these factors will be of help in planning suitable disability-management packages, including the rehabilitation of those who find it difficult to carry on with their existing jobs because of the severity of their disease. PMID- 14754497 TI - The effectiveness of 3, 5 or 7 days of albendazole for the treatment of Trichuris trichiura infection. AB - A randomized clinical trial was carried out to study the relationship between the duration of albendazole therapy, at 400 mg/day, and its effectiveness in the treatment of Trichuris trichiura infection. The 168 patients were treated for three (N=56), five (N=56) or seven (N=56) consecutive days. Compared with both of the shorter regimens, treatment for 7 days resulted in a significantly higher cure 'rate' and significantly greater reductions in the level of egg excretion. The advantage of using the longer (5- or 7-day) regimens was most apparent among the patients who had heavy infections (at least 1000 Trichuris eggs/g faeces) when treated. It is therefore suggested that albendazole be given for at least 3 days to those with light infections and for 5-7 days to patients with heavy infections. PMID- 14754498 TI - Strongyloides hyper-infection: a case for awareness. AB - In patients receiving immunosuppressive therapies, Strongyloides stercoralis can cause a life-threatening septic shock, with multi-organ failure and infestation. Strongyloides hyper-infection should be considered in any immunosuppressed patient who has been exposed to the parasite, even if it is many years since that exposure occurred. Delayed eosinophilia may be a feature and treatment with high doses of anthelmintics may be required. An interesting case of S. stercoralis hyper-infection was recently observed at the Royal Darwin Hospital in tropical, northern Australia. The patient was an 18-year-old female with lupus glomerulonephritis, who was receiving immunosuppression in the form of corticosteroids and pulse cyclophosphamide. The characteristics and intensive care management of this case, including the use of granulocyte-colony stimulating factor and high-dose ivermectin, are described. The patient, who survived, appears to represent the first reported case of S. stercoralis hyper-infection with suspected myocarditis. PMID- 14754499 TI - A sero-epidemiological study of Rickettsia typhi infection in dogs from Soria province, central Spain. AB - Data relating to Rickettsia typhi infection in Spain are scarce. The seroprevalence of canine infection with this species has now been investigated, in dogs from the central province of Soria. The results of indirect immunofluorescence assays indicated that nine (12.3%) of the 73 dogs checked were carrying antibodies against R. typhi. The gender, age and breed of the dog, and whether it was used for hunting, shepherding, guarding or simply as a pet, apparently had no significant affect on the probability of it being seropositive. Being infested with fleas or having a history of such infestation was, however, significantly associated with seropositivity. The present results confirm that dogs may be infected with R. typhi and indicate their potential usefulness as sentinels in epidemiological studies of the pathogen. The results of wide-scale, serological studies of dogs may allow the geographical distribution of R. typhi to be mapped relatively simply. PMID- 14754500 TI - Acaroid mites (Acari: Astigmata) in Chinese traditional medicines. AB - Almost 2000 samples of the dried plants and animals used in Chinese traditional medicine were collected from a storehouse in Huainan city, Anhui province, China, where they had been kept for at least 6 months. When examined, every sample was found to contain acaroid mites. The mites included representatives of 44 species belonging to 22 genera and seven families. It seems that samples of Chinese traditional medicine frequently become severely infested with mites while in storage. More importance should be attached to protecting the medicines against mite infestation and to protecting against acariasis those who ingest or simply handle the medicines. PMID- 14754501 TI - Placental Plasmodium vivax infection and congenital malaria in central India. PMID- 14754502 TI - Fibrinopeptide-A and fibrinogen interactions in acute, Plasmodium falciparum malaria. PMID- 14754503 TI - Involvement of protein kinase C-delta in DNA damage-induced apoptosis. AB - Apoptosis is a highly orchestrated cell suicidal program required to maintain a balance between cell proliferation and cell death. A defect in apoptotic machinery can cause cancer. Many anticancer drugs are known to kill tumor cells by inducing apoptosis, and a defect in apoptosis can lead to anticancer drug resistance. Apoptosis is regulated by a complex cellular signaling network. Several members of the protein kinase C (PKC) family serve as substrates for caspases and PKCdelta isozyme has been intimately associated with DNA damage induced apoptosis. It can act both upstream and downstream of caspases. In response to apoptotic stimuli, the full-length and the catalytic fragment of PKCdelta may translocate to distinct cellular compartments, including mitochondria and the nucleus, to reach their targets. Both activation and intracellular distribution of PKCdelta may have significant impact on apoptosis. This review intends to assimilate recent views regarding the involvement of PKCdelta in DNA damage-induced apoptosis. PMID- 14754504 TI - Endoplasmic reticulum Ca(2+) homeostasis and neuronal death. AB - The endoplasmic reticulum (ER) is a universal signalling organelle, which regulates a wide range of neuronal functional responses. Calcium release from the ER underlies various forms of intracellular Ca(2+) signalling by either amplifying Ca(2+) entry through voltage-gated Ca(2+) channels by Ca(2+)-induced Ca(2+) release (CICR) or by producing local or global cytosolic calcium fluctuations following stimulation of metabotropic receptors through inositol 1,4,5-trisphosphate-induced Ca(2+) release (IICR). The ER Ca(2+) store emerges as a single interconnected pool, thus allowing for a long-range Ca(2+) signalling via intra-ER tunnels. The fluctuations of intra-ER free Ca(2+) concentration regulate the activity of numerous ER resident proteins responsible for post translational protein folding and modification. Disruption of ER Ca(2+) homeostasis results in the developing of ER stress response, which in turn controls neuronal survival. Altered ER Ca(2+) handling may be involved in pathogenesis of various neurodegenerative diseases including brain ischemia and Alzheimer dementia. PMID- 14754505 TI - Plasticity of purine release during cerebral ischemia: clinical implications? AB - Adenosine is a powerful modulator of neuronal function in the mammalian central nervous system. During a variety of insults to the brain, adenosine is released in large quantities and exerts a neuroprotective influence largely via the A(1) receptor, which inhibits glutamate release and neuronal activity. Using novel enzyme-based adenosine sensors, which allow high spatial and temporal resolution recordings of adenosine release in real time, we have investigated the release of adenosine during hypoxia/ischemia in the in vitro hippocampus. Our data reveal that during the early stages of hypoxia adenosine is likely released per se and not as a precursor such as cAMP or an adenine nucleotide. In addition, repeated hypoxia results in reduced production of extracellular adenosine and this may underlie the increased vulnerability of the mammalian brain to repetitive or secondary hypoxia/ischemia. PMID- 14754506 TI - Human natural killer cell receptors: insights into their molecular function and structure. AB - NK cells express receptors characterized by opposite functions that finely regulate their activities. Among inhibitory receptors, some are specific for different groups of MHC class I alleles, while others are still orphan receptors. On the contrary, various activating receptors are involved in the triggering of NK-mediated natural cytotoxicity. In general, their engagement induces human NK cells to kill target cells that are either HLA class I-negative or -deficient. Thus, the process of NK cell triggering mediated by Natural Cytotoxicity Receptors can be mainly considered as a non MHC-restricted mechanism. Here, a brief description of the molecular nature of these receptors, as well as, of their 3D-structures and of the implications for ligand recognition, is given. PMID- 14754507 TI - Peroxisome biogenesis and the role of protein import. AB - Peroxisomes are metabolic organelles with enzymatic content that are found in virtually all cells and are involved in beta-oxidation of fatty acids, hydrogen peroxide-based respiration and defence against oxidative stress. The steps of their biogenesis involves "peroxins", proteins encoded by PEX genes. Peroxins are involved in three key stages of peroxisome development: (1). import of peroxisomal membrane proteins; (2). import of peroxisomal matrix proteins and (3). peroxisome proliferation. Of these three areas, peroxisomal matrix-protein import is by far the best understood and accounts for most of the available published data on peroxisome biogenesis. Defects in peroxisome biogenesis result in peroxisome biogenesis disorders (PBDs), which although rare, have no known cure to-date. This review explores current understanding of each key area in peroxisome biogenesis, paying particular attention to the role of protein import. PMID- 14754508 TI - Ischemia induces short- and long-term remodeling of synaptic activity in the hippocampus. AB - One of the most vulnerable areas to ischemia or hypoglycemia is CA1 hippocampal region due to pyramidal neurons death. Glutamate receptors are involved together with protein-kinase C and nitric oxide synthase. Long-term potentiation (LTP) is generated in anoxic or hypoglycemic conditions via activation of NMDA while inhibition of these receptors attenuates this response. Protein-kinase C and nitric oxide synthase are involved in anoxic LTP mechanism. Postischemic neurons are hyperexcitable in CA3 area while CA1 pyramidal neurons degenerate and disappear. Changes of glutamate receptors triggered by ischemia and hypoglycemia are discussed in this review. PMID- 14754509 TI - Genetic clock of biologic rhythms. AB - Biologic rhythms are tightly regulated in all species, in order to adapt to circadian variations of light, temperature and other biological related living conditions. The intrinsic mechanisms of rhythm regulation from every cell to a whole organism is embedded into the genetic clockwork. The mammalian clock genes are reviewed with their interrelation. The per gene promoter is stimulated by clock/bmal1 heterodimer and trigger the per proteins synthesis. Initially considered as a simple negative feed-back, from to the phosphorylated per/cry heterodimer towards clock/bmal1 transcription stimulus, the clockweb of the cell became complex. General characteristics of cell clock genes are integrated in a model. In addition, the involvement of clock genes in peripheral tissues is discussed. PMID- 14754510 TI - Endotoxin translocation in two models of experimental acute pancreatitis. AB - To test the hypothesis that endotoxin is absorbed from the gut into the circulation in rats with experimental acute pancreatitis we studied two different animal models. In the first model necrotizing pancreatitis was induced by the ligation of the distal bilio-pancreatic duct while in the second, experimental oedematous acute pancreatitis was induced by subcutaneous injections of caerulein. In both experiments, in the colon of rats with acute pancreatitis endotoxin from Salmonella abortus equi was injected. Endotoxin was detected by immunohistochemistry in peripheral organs with specific antibodies. The endotoxin was found only in rats with both acute pancreatitis and endotoxin injected into the colon and not in the control groups. The distribution of endotoxin in liver at 3 and 5 days was predominantly at hepatocytes level around terminal hepatic venules, while in lung a scattered diffuse pattern at the level of alveolar macrophages was identified. A positive staining was observed after 12 hours in the liver, lung, colon and mesenteric lymph nodes of rats with both caerulein pancreatitis and endotoxin injected into the colon. We conclude that the experimental acute pancreatitis leads to early endotoxin translocation from the gut lumen in the intestinal wall and consequent access of gut-derived endotoxin to the mesenteric lymph nodes, liver and lung. PMID- 14754511 TI - Phenotypic characterization of mouse embryonic fibroblasts lacking heat shock factor 2. AB - In murine cells, the heat shock response is regulated by a transcription factor, HSF1, which triggers the transcription of heat shock genes. HSF2 has been shown to be involved in meiosis and mouse brain development. We characterized the effects of the absence of HSF2 in mouse embryonic fibroblasts (MEFs). The temperature threshold of the heat shock response appeared lowered in Hsf2(-/-) MEFS as monitored by the synthesis of heat shock protein HSP70. In contrast to unstressed wild type MEFS, HSP70 and HSF1 are localized in the nucleus of unstressed Hsf2(-/-) MEFS, a characteristic of stressed cells. HSF1 is not activated for DNA-binding at unstressed temperature in Hsf2(-/-) MEFS. Therefore, the absence of HSF2 induces some but not all of the characteristics of the stress response. In addition, Hsf2(-/-) MEFS exhibited proliferation defects, altered morphology, remodeling of the fibronectin network. PMID- 14754512 TI - Immunohistochemical assessment of proliferating cell nuclear antigen in primary hepatocellular carcinoma and dysplastic nodules. AB - A complementary way for the assessment of HCC prognosis is represented by the analysis of molecular markers. Thus, immunohistochemical assessment of proliferation can describe tumor aggressiveness, probability of local recurrence or metastasis potential, being very useful for the assessment of recurrence-free survival and survival until death. The aim of our study was to assess proliferating cell nuclear antigen activity in HCC and dysplastic nodules as compared with surrounding non-neoplasic areas. Immunohistochemical techniques were thus performed on the samples obtained by ultrasound-guided liver biopsies or intraoperative biopsies, in 32 patients with HCC, as well as in 3 patients with dysplastic nodules occurring in liver cirrhosis. Expression of PCNA within extranodular areas of the HCC patients in the absence or presence of cirrhosis, was increasing from 40% to 70%, respectively. PCNA expression further increased within intranodular areas of dysplastic nodules and HCC, to 100% and 96.88%, respectively. A progressive increase of the mean values of PCNA-LI was also observed from extranodular areas without or with cirrhosis, towards intranodular areas of dysplastic nodules and HCC (4.2%, 6.8%, 27.9%, 31.9%, respectively). Dysplastic nodules can thus be considered lesions with a high-proliferation rate, representing an early stage of hepatocarcinogenesis. This supported the current recommendations for borderline hepatocellular nodules identified by ultrasound, which indicate an aggressive treatment similar to malignant lesions. In summary, we demonstrated a progressively increasing rate of cellular proliferation, from extranodular non-neoplasic areas to intranodular areas (dysplastic nodules and HCC), as reflected by an increased expression of proliferating cell nuclear antigen labelling index. PMID- 14754513 TI - The influence of sodium metavanadate on the process of diabetogenesis in BB rats. AB - Vanadium has been shown to be beneficial in the oral treatment of animal models of type 1 and type 2 diabetes. The aim of the study was to evaluate the short term effects of sodium metavanadate in prediabetic BB-DP rats. To do this, 96 rats were divided into 4 equal groups. Groups V1, V2, V3 were treated with sodium metavanadate (0.1, 0.2 and 0.3 mg/ml respectively) and sodium chloride (0.5 mg/ml) in drinking water for 7 days. Group C received only sodium chloride (0.5 mg/ml). Blood glucose (BG), glycosuria, ketonuria, body weight and insulinemia were determined. The age of onset of diabetes was significantly higher for groups V2, V3 compared to group C, (p<0.05) and depends on the metavanadate concentration (V3 vs. V1, p=0.006). The incidence of diabetes was lower in the rats treated with metavanadate than in the control group, but this difference was not statistically significant. In diabetic rats, the BG at the onset was higher in group C than in groups V, p<0.05. Insulinemia, at the onset of the treatment as well as immediately after its cessation showed a drop in the treatment groups, proportionally to the dosage of vanadium, but later increased slowly and continuously until the end of the experiment. In conclusion, metavanadate delays the development of diabetes in BB-DP rats, but does not prevent its onset. A milder form of diabetes occurs in diabetic rats treated with metavanadate. The effects depend on the metavanadate concentration and 0.2 mg/ml is preferable. PMID- 14754514 TI - Endothelial cells from hematopoietic stem cells are functionally different from those of human umbilical vein. AB - Hematopoietic stem cells have a remarkable plastic capacity, which allows them to differentiate into various cells, such as immune cells, nervous cells, muscle cells, bone and cartilaginous cells. The aim of this study was to show the capacity of stem cells to differentiate into endothelial cells, in culture, after addition of endothelial cells growth supplement (ECGS). We also compared the behavior of these cells with that of endothelial cells obtained from human umbilical vein (HUVEC). CD34+ cells obtained by immunomagnetic separation from human umbilical cord and placental blood were used. After 12-15 days of culture in a medium containing ECGS, the cells showed morphological changes characteristic to endothelial cells and immunocytochemical analysis revealed the presence of CD31 surface antigen and von Willebrand factor. The flow-cytometric analysis of endothelial cells adhesion molecules (ECAM) showed that endothelial cells derived from CD34+ cells expressed CD54/ICAM-1 9.65+/-0.2% and CD106/VCAM 7.73+/-0.3%, values similar to those expressed by HUVECs. After TNF incubation, ECAM expression increased only in HUVECs. These data demonstrate that a fraction of circulating CD34+ cells may develop some endothelial cell characteristics when cultured with ECGS, but they are functionally different from HUVECs. PMID- 14754515 TI - Sertoli-Leydig cell tumor - a rare androgen secreting ovarian tumor in postmenopausal women. Case report and review of literature. AB - Sertoli-Leydig cell tumors (SLCT) constitute only 1-0.5% of all primary ovarian neoplasms. We report a SLCT in a postmenopausal woman aged 69 years. The physical examination revealed severe hirsutism. Basal hormonal evaluation showed high plasma testosterone and estradiol values, with suppressed plasma gonadotropins. Computer tomograph scan revealed a right ovarian tumor mass of 4,3/3 cm, confirming an androgen secreting ovarian tumor. The histopathological and immunocytochemical examination established the diagnosis of well differentiated Sertoli-Leydig cell tumor. The tumor was positive for cytokeratin KL 1 and S-100 protein and, in isolated tumor cells, positive for alpha-fetoprotein. Postsurgical evolution was favorable; controls after 6 months and 3,5 years showed marked reduction of hirsutism, normal plasma testosterone values and gonadotropins in normal postmenopausal range. We discuss the complex aspects of etiology and pathogenesis, the clinical and hormonal settings, the role of immunocytochemical markers in diagnosis, as well as the therapy and the prognostic features of this ovarian tumor. PMID- 14754516 TI - Interleukin-17 in acute myeloid leukemia. AB - There are several reports that angiogenesis plays important roles in hematological malignancies including acute myeloid leukemia (AML). Human interleukin-17 (IL-17) is a proinflammatory cytokine produced by activated CD4 T cells. IL-17 plays a potential role in T cell mediated angiogenesis. The role of IL-17 in pathologic angiogenesis has not been evaluated yet. The aim of the study was to determine plasma level of IL-17 in patients with AML. IL-17 levels were measured by ELISA in plasma samples taken from 68 adult patients with AML before chemotherapy was administered. In addition 20 out of 68 patients were reanalysed after achieving complete remission (CR). Ten samples from healthy volunteers were evaluated as the control. In this study we have demonstrated that serum level of IL-17 is not elevated in AML patients. These results suggest that angiogenesis in AML is not mediated by CD4 T cells. To our knowledge this is the first report about IL-17 serum level in acute leukemias. We are currently evaluating IL-17 levels in others haematological malignancies. PMID- 14754517 TI - New asthma drugs acting on gene expression. AB - New asthma drugs acting on transcription are transcription factor agonists (dissociated steroids, peroxisome proliferator-activated receptor gamma agonists), transcription factor inhibitors (NF-kappaB / AP-1 inhibitors, STAT6 inhibitors), inhibitors of protein kinases acting on transcription factors (p38 MAP kinase inhibitors), and chromatin modifying agents. Pharmacological approach of translation in asthma includes therapeutic ribozymes and antisense oligonucleotides targeting receptors (adenosine A1 receptor, alpha chain of IL-5 receptor, common beta chain of IL-3/IL-5/GM-CSF receptor), cytokines (IL-4, IL-5, SCF), signal transduction molecules (Syk, Lyn), transcription factors (STAT-6, GATA-3). Some of these drugs acting on gene expression have the potential to improve therapeutic benefits compared with traditional drugs. PMID- 14754519 TI - Glomerular developmental chronology in human fetuses. PMID- 14754518 TI - Focal steatosis of the liver with metastatic carcinoma. PMID- 14754520 TI - Molecular pathology in the diagnosis and treatment of non-Hodgkin's lymphomas. AB - The non-Hodgkin's lymphomas encompass a wide spectrum of hematologic neoplasms that exhibit different clinical and biological features. Lymphomas classically have been initially assessed based on their cytologic and histologic features. Morphology alone is often inadequate as similar appearing neoplasms may be immunophenotypically and molecularly heterogeneous. Molecular diagnostic methods can provide an additional level of testing that not only helps refine diagnoses but can provide prognostic information. New methods are being refined that may provide information to establish precise diagnostic profiles, provide targets for therapy and provide more sensitive methods for monitoring the success of treatment. Molecular methods will be increasingly utilized and eventually required as the accepted method of diagnosis and for monitoring the disease. Understanding of the molecular abnormality and the pathogenesis of the neoplasm hopefully will lead to therapeutic intervention aimed at the specific molecular defect or its product. The molecular pathology of the non-Hodgkin's lymphomas is discussed. PMID- 14754522 TI - [Putting emphasis on the prevention and management of diabetes in children and adolescents]. PMID- 14754521 TI - [Diagnosis and treatment of inherited metabolic diseases: past and future]. PMID- 14754523 TI - [Application of gas chromatography-mass spectrometry analysis on urine filter paper in the high-risk screening and diagnosis of inherited metabolic diseases]. AB - OBJECTIVE: To establish a specific procedure for the high-risk screening and diagnosis of organic acidurias and other inherited metabolic diseases in China. METHODS: A nation-wide network for the high-risk screening and diagnosis of genetic metabolic diseases was formed to facilitate the collaboration. Urine samples were collected using filter paper from patients with clinical symptoms suspicious of inherited metabolic diseases. The samples were eluted with distilled water and internal standards were added. Samples were treated with hydroxylamine hydrochloride to form oximes to improve the recoveries of 2 ketoacids. Urinary organic acids were extracted with ethyl acetate and diethyl ether under acidic condition. After dehydration, the combined organic phase was evaporated to dryness with nitrogen. The residues were added with BSTFA + 1%TMCS and heat incubated to form the trimethylsilyl derivatives, and then were analyzed on an Agilent 5890/5973N gas chromatography-mass spectrometer (GC-MS), with a 7683 series auto-sampler. The peaks were identified by reference to a mass spectral library. RESULTS: Totally 352 samples were collected from the network collaborating hospitals since 2001. Thirty-four (9.66%) cases of various inherited metabolic diseases were diagnosed with an age range of 2 days to 14 years. The disease profile was consisted of methylmalonic acidemias (6), alpha keto-glutaric aciduria (5), tyrosinemia type I (4), dicarboxylic aciduria (4), multiple carboxylase deficiency (3), phenylketonuria (3), lactic acidemia (3), propionic acidemia (2), ornithine transcarbamoylase deficiency (1), ethylmalonic adipic aciduria (1), glutaric aciduria type II (1) and 3-methylcrotyl CoA carboxylase deficiency (1). The most common clinical symptoms and signs included mental and developmental retardation, convulsion, musculotonic abnormality and jaundice. Routine laboratory tests often revealed metabolic acidosis, hypoglycemia and hyperammonemia, etc. CONCLUSION: Urine organic acids analysis by GC-MS remains to be the most important technique for the high-risk screening and diagnosis of inherited metabolic diseases. Use of urine filter paper for sample collection and analysis in advanced genetic metabolic centers is a practical approach to extend the diagnostic capacity and improve the management of such diseases in China. Collaborative network played a critical role in the success of the program. PMID- 14754524 TI - [Diagnosis and treatment of biotinidase deficiency-clinical study of six patients]. AB - OBJECTIVE: To investigate the clinical and neurodevelopmental profiles of patients with biotinidase deficiency and to determine the efficacy of current therapy with respect to outcome. METHODS: Six patients aged from 3 months to 14 years with biotinidase deficiency were confirmed by urinary organic acid analysis with gas chromatography/mass spectrometry (GC/MS) and biotinidase assay on dried blood spots. Biotin was supplemented individually (10-40 mg/d). Their clinical features, laboratory findings, and treatment regimen were reviewed. RESULTS: All the 6 patients presented with some extent of neurological abnormalities and dermatological lesions. Cases 1 - 3 had poor feeding, vomiting, seizures, mental retardation, and lethargy onset from their early infancy, with varied degree of anemia, ketosis, acidosis, and hypoglycemia. Case 2 exhibited eczema and dermatitis from his age of 7 months. Case 4 displayed motor deficit and ataxia after 6 months of age, and generalized pustular psoriasis when he was 8 months old. Cases 5 and 6 gradually showed muscle weakness and paraplegia at the age of 7 years and 5 years, respectively. Inflammatory demyelination changes of cervical cord were evident on magnetic resonance imaging in these two patients. Case 6 had progressive optic atrophy, eczema and alopecia. Remarkable elevations of urinary lactate, pyruvate, 3-OH-propionate, methylcitrate, propionylglycine, 3-OH isovalerate, 3-methylcrontonylglycine were confirmed in cases 1, 2, 3 and 5. Slight increase of urinary lactate, pyruvate, and 3-methylcrontonylglycine was observed in cases 4 and 6. Biotinidase activities assayed on dried blood spots from all the patients were below 0.1 pmol/(min.3 mm) Biotin supplementation for all the patients, except for case 3 who was not treated, resulted in pronounced and rapid clinical and biochemical improvement. Cases 4 and 6 had residual neurological damage comprising ataxia and motor handicap of legs, due to prolonged disease course. CONCLUSIONS: Biotinidase deficiency intensively impairs nervous system and skin in the affected patients. Urinary organic acid analysis and blood biotinidase assay are crucial to the diagnosis. Early diagnosis and biotin supplementation can contribute significantly to the improvement of prognosis. PMID- 14754525 TI - [Heterogeneous phenotypes in Chinese glycogen storage disease type Ia patients with homozygous G727T mutation]. AB - Glycogen storage disease (GSD) type Ia is an autosomal recessive disorder caused by a deficiency of glucose-6-phosphatase (G6Pase). The gene that encodes G6Pase was mapped to 17q21. The molecular genetic basis of GSD type Ia in the mainland Chinese population has not been explored. OBJECTIVE: To analyze the G6Pase gene mutations, and to compare the phenotypic features and the response to the corn starch treatment among patients who share the same mutation. METHODS: With the consent of the parents and their children, the authors studied 18 families with clinically diagnosed GSD type Ia from our long time follow-up groups. Direct DNA sequencing of all 5 exons and the exon-intron boundaries of G6Pase gene were done on the blood specimens. Seven of the 18 patients, male 2 and female 5, aged 1.5 to 16 years, were homozygous for same mutation. The clinical symptoms, signs and the serum biochemical values before and after treatment were compared in these 7 patients. RESULTS: The 7 patients were homozygous of G-->T transversion at the nucleotide 727 in exon 5 (G727T), which has previously been reported to cause abnormal splicing. The parents were heterozygous of the G727T mutation. All the patients exhibited typical features of GSD type Ia with variable severity, including hypoglycemia, hepatomegaly, kidney enlargement, growth retardation, bleeding diathesis, lactic acidemia, hyperlipidemia, and hyperuricemia. Two of the patients had repeated hypoglycemic seizures before the age of 2 years. One had moderate splenomegaly when he came to our clinic at the age of 16, the spleen size was reduced to 2 cm below the left costal margin after 5-year treatment. His sister, homozygous of G727T, did not show splenomegaly. One had multiple hepatic adenoma since the age of 5 years. Four had 5-year-delayed bone age when they started treatment at the age of 9 to 16 years, the bone age reached normal after 2 - 3 years treatment. No matter when they started corn starch treatment, the height increase in the first year was most obvious with an average of 10 cm. All the patients had abnormal liver function before treatment, 5 had constant slightly elevated liver enzymes with the treatment. All had normal urinalysis test, but the urine beta(2)- microglobulin was elevated. CONCLUSIONS: G727T mutation may be the major cause of GSD type Ia in China. Patients with the same mutation could have variable phenotypic characteristics, and the response to the corn starch treatment was different. The diagnosis of GSD type Ia can be based on clinical and biochemical abnormalities combined with mutation analysis instead of enzyme assays on liver biopsy. PMID- 14754526 TI - [Gene diagnosis of X-linked spondyloepiphyseal dysplasia tarda by linkage analysis and DNA sequencing]. AB - OBJECTIVE: X linked spondyloepiphyseal dysplasia tarda (SEDL) is heritable osteochondrondysplasia characterized in affected males by disproportional short stature with short neck and trunk resulting from a growth defect of the vertebral bodies, accompanied by barrel chest and degenerative osteoarthropathy of hip joints. This progressive skeletal dysplasia is caused by the SEDL gene located approximately 100 kb centromeric of DXS16 at Xp22. The disorder usually manifests in late childhood without systemic complications, and generally female carriers of SEDL are asymptomatic. So the diagnosis of potential carriers and presymptomatic patients is almost impossible. This study aimed to establish methods of gene diagnosis for finding out potential carriers and presymptomatic patients. METHODS: The blood samples were collected from 21 individuals in a large Chinese pedigree with SEDL. Microsatellite marker DXS16 was selected for linkage analysis. In order to confirm the allele of DXS16 linked to the pathogenic SEDL gene, polymerase chain reaction (PCR) and polyacrylamide gel electrophoresis (PAGE) were used to examine the variability of the lengths of DXS16, and linkage analysis was performed for the diagnosis of potential carriers and presymptomatic patients. Then the pathogenic mutation of the SEDL gene in the family was identified by bi-directionally direct sequencing of PCR products amplified for each of the four coding exons as well as their exon/intron boundaries. The potential carriers and presymptomatic patients were also diagnosed in this way. RESULTS: Six young individuals (IV(14), IV(19), IV(21), IV(23), V(4), V(7))who wanted to know whether they were carriers or presymptomatic patients were diagnosed by linkage analysis. Four females of them (IV(14), IV(19), IV(21), V(7)) were determined being carriers because they carry the allele of DXS16 which links the pathogenic SEDL gene, and the other two (IV(23), V(4)) being normal individuals for their alleles of DXS16 linked with wild SEDL gene. DNA sequencing identified that the pathogenic mutation of SEDL gene in the family, which was a nucleotide substitution of the splice-acceptor site in intron 2, IVS2 -2 A-->C. This is a novel mutation in the SEDL gene. Four female individuals (IV(14), IV(19), IV(21), V(7)) carried the mutation; individuals IV(23) and V(4) carried the wild SEDL gene. The results of diagnosis of linkage analysis coincide completely with that of DNA sequencing. CONCLUSION: Linkage analysis is a simple, rapid and inexpensive gene diagnosis method for SEDL and its accuracy was the same as DNA sequencing. Each of linkage analysis and DNA sequencing can be used to diagnose SEDL, which is very helpful for finding potential carriers and presymptomatic patients. PMID- 14754528 TI - [Management of inherited metabolic diseases]. PMID- 14754527 TI - [Association of human leukocyte antigen non-classical genes with type 1 diabetes]. AB - OBJECTIVE: HLA-DMA and DMB are non-classical genes whose product (DM molecules) plays an important role in antigen presentation. Our present study was designed to investigate the relationship between human leukocyte antigen-DMA, -DMB and clinical status heterogeneity of type 1 diabetes. METHODS: A total of 80 children (male 36, female 44) with type 1 diabetes were selected as research subjects. Diagnosis of type 1 diabetes was made according to WHO criteria. The range of age at onset of type 1 diabetes was 2.5 - 14 years. Ninety-one healthy adult blood donors were selected as normal controls. Polymerase chain reaction and dot blot hybridization techniques were used to classify DMA and DMB alleles. Patients with type 1 diabetes were classified into different groups according to different clinical status, including sex, age of onset, ketosis onset situation on diagnosis, remained function of islet beta cell, etc. Then distribution of DM susceptive alleles and heterodimer in different clinical groups were studied. RESULTS: The frequencies of DMA * 0103 and DMB * 0103 alleles in patients were significantly increased (50% vs. 8%, 43% vs. 22%, respectively), these two alleles confer susceptibility to type 1 diabetes in Chinese. The frequencies of DMA * 0103/DMB * 0102, DMA * 0103/DMB * 0103 and DMA * 0103/DMB * 0101 heterodimers were also increased in the patients. The above heterodimers confer predisposition to type 1 diabetes. Both DMB * 0103 allele and DM susceptive heterodimers are related to islet beta cell function on diagnosis. The patients with DMB * 0103 allele or DM susceptive heterodimers were significantly increased in the patients with lower C-peptide level on diagnosis (56% vs. 29%; 58% vs. 34% respectively). DM heterodimes were also related to onset age and ketosis-onset situations of the patients. The patients carrying DM susceptive heterodimers had higher probability to suffer type 1 diabetes before 10 years of age and had the predisposition to ketosis or ketoacidosis on diagnosis. CONCLUSION: HLA- class II non-classical alleles-DMA and DMB may play an important role in pathogenesis of type 1 diabetes, and clinical status heterogeneity of type 1 diabetes may be related to genetic mechanism. PMID- 14754530 TI - [Misdiagnosis of acute intermittent porphyria in a case]. PMID- 14754529 TI - [Type 2 diabetes in children and adolescents]. PMID- 14754531 TI - [Recommendations for the diagnosis and treatment of central precocious puberty]. PMID- 14754532 TI - [Report of a case with Rotor syndrome]. PMID- 14754533 TI - [Diagnostic study on children's diaphragmatic fatigue]. AB - OBJECTIVE: In the recent twenty years, the diaphragmatic contraction, relaxation functions and electric activity have been explored through electromyography (EMG) and transdiaphragmatic pressure (Pdi) determination. But these techniques required some complex and expensive instruments, so the diagnosis and treatment of children's diaphragmatic fatigue have not been well evaluated. The present study explored the diagnosis of children's diaphragmatic fatigue through measuring ribcage-abdomen motion and analyzed its asynchrony. METHODS: Fifty three children (male 37, female 16, and age rage from 1 months to 9 years) with respiratory rate > 30 breaths/min, heart rate > 110 beats/min, and respiratory dysfunction had asynchronized ribcage-abdomen motion showed by impedance respirograph (IRG). The authors observed whether ribcage-abdomen motion was synchronic and calculated M levels (staggered peak time/total duration of the breathing cycle). The ribcage and abdomen outputs were displayed on vertical (for rib cage) and horizontal (for abdomen) axes of X-Y instrument. In addition, the change of respiratory frequency and heart rate was observed and arterial blood gas analysis was also performed. RESULTS: (1) M levels in one-dimensional IRG were positively correlated with alpha angle in two-dimensional IRG (r = 0.956, P < 0.001). Asynchronized respiratory motions could be divided into three types. type I showed completely contra-directional movements of respiration, M levels for (48.1 +/- 4.4)%, an irregularly clockwise loop in the two dimensional IRG, and alpha angle for (138.3 +/- 15.0) degrees. In type II, one dimensional IRG showed displaced peak of the chest and abdomen motion curves, M levels were (16.5 +/- 4.7)%, two dimensional IRG was displaced in a counterclockwise direction, and alpha angle was (55.3 +/- 10.8) degrees. In type III, abdominal motion curve of one dimensional IRG had double peaks, M levels were 0, two dimensional IRG was presented as 8-shaped double circles, alpha angle was (41.3 +/- 3.8) degrees; (2) pH levels in the patients with type I and type II diaphragmatic fatigue were significantly lower, and PCO(2) levels were significantly higher than those with type III or in the normal subjects (P < 0.001 for all), but there was no statistically significant difference between type III and the normal subjects (P > 0.05); (3) Both of respiratory rate and heart rate in type I, type II and type III were higher than those in the normal subjects (all P < 0.001), and the differences among the three types were significant (P < 0.001 for all); (4) Both M levels and alpha angle were negatively correlated with pH levels (r = -0.514, P < 0.001 and r = -0.497, P < 0.001), while positively correlated with PCO(2) levels (r = 0.672, P < 0.001 and r = 0.625, P = 0.01). CONCLUSIONS: (1) IRG can be reliably used to diagnose children's diaphragmatic fatigue. This technique is simple and easy to perform and non-invasive. It is therefore worthy of recommending for further clinical investigations. (2) According to the characteristics of IRG, diaphragmatic fatigue can be divided into three types. (3) The development of children's diaphragmatic fatigue has a series of characteristic changes. (4) To avoid the patients suffering from respiratory failure, it is the key time to adopt the policies of prevention and treatment when IRG shows signs of type III diaphragmatic fatigue. PMID- 14754534 TI - [Eosinophils apoptosis in asthmatic children]. AB - Prominent eosinophil airway inflammation is important in the pathogenesis of asthma. There is increasing evidence that the disorder of eosinophil apoptosis contributes to the mechanism. But most of the studies have been done in vitro or on animal models, very few were done among the adult asthmatics in vivo. OBJECTIVE: The aim of this study was to elucidate the relationship between the apoptotic eosinophils and Bcl-2 in asthmatic children in vivo. METHODS: Eleven mild to moderate asthmatic patients were recruited and the range of age was 7 - 14 years (9 males, 2 females), meanwhile 7 patients with lower respiratory infection were recruited as control and the range of age was 9 - 14 years (5 males, 2 females). Before and after inhaled glucocorticoid (GC) induced sputum, bronchoalveolar lavage (BAL), bronchial mucosa specimens and peripheral blood were obtained for measuring and comparing the changes of apoptotic EG(2)(+) cell by combining the techniques of TUNEL and immunohistochemistry, meanwhile the expression of Bcl-2 in bronchial mucosa specimens was measured by using the immunohistochemical assay. RESULTS: Before the inhalation of GC, the apoptotic EG(2)(+) cells in asthmatics were significantly lower than that in control group (P < 0.01), and the numbers of EG(2)(+) cell in asthmatics group were significantly higher than that in control group (P < 0.001). After the treatment apoptotic EG(2)(+) cells in asthmatics were increased (P < 0.01), and the numbers of EG(2)(+)cell were decreased (P < 0.01, P < 0.05 and P < 0.05, respectively), FEV(1)% was increased (P < 0.05). Before the inhalation of GC, the numbers of Bcl 2(+) cell in asthmatic airway submucosa were higher than that in control group (P < 0.05) but after the treatment the number of Bcl-2(+) cell did not change significantly. (4) Before and after GC treatment the percentages of apoptotic eosinophils of peripheral blood in vivo had no significant changes compared with those of control subjects (P > 0.05). There was a positive correlation between apoptosis of EG(2)(+) cell in sputum, BAL, airway submucosa and FEV(1)% (P < 0.05). CONCLUSION: Apoptosis of EG(2)(+) cell decreased in the airway of asthmatic children and inducing EOS apoptosis is one of the important mechanism of inhaled GC therapy for asthma. PMID- 14754535 TI - [Effect of dexamethasone, aminoguanidin, amrinone on oxygen utilization in endotoxin shock rabbits]. AB - OBJECTIVE: To investigate the effect of three kinds of drug with different mechanism, dexamethasone (Dex), aminoguanidin (AG) and amrinone (Amr) on oxygen utilization in endotoxic shock rabbits. METHODS: Thirty-five rabbits were randomly allocated into five groups: operation, lipopolysaccharide (LPS), Dex, Amr and AG. The endotoxin shock was induced by intravenously injecting LPS (400 micro g/kg). The arterial blood gas, mixed venous blood gas and cardiac output were recorded at 30 min after the operation (T(0)), shock status (T), 1 - 6 h after the treatment (T(1)-T(6)). The oxygen delivery (DO(2)), oxygen consumption (VO(2)) and extraction ratio of oxygen (ERO(2)) were calculated. RESULTS: All the parameters in five groups showed no significant differences (P > 0.05) at T(0). Six hours after treatment, rabbits in Dex group presented with significantly improved DO(2) (12.4 +/- 3.1) ml/(kg.min), P < 0.01 and VO(2) (5.1 +/- 1.6) ml/(kg.min), P < 0.05 compared with DO(2) (8.1 +/- 2.4) ml/(kg.min) and VO(2) (2.7 +/- 1.0) ml/(kg.min) in LPS group. Rabbits in AG group showed significantly increased DO(2) (17.0 +/- 2.8) ml/(kg.min) (P < 0.01), (17.2 +/- 2.5) ml/(kg.min) (P < 0.05), compared with (12.2 +/- 2.6), (14.1 +/- 3.8) ml/(kg.min) in LPS group at T(1) and T(2), respectively, but there was no significant difference (11.2 +/- 1.7) ml/(kg.min) (P > 0.05) at T(6). The VO(2) increased significantly, (5.0 +/- 1.0) ml/(kg.min) (P < 0.01) compared with LPS group at T(6). The VO(2) of Amr group was significantly higher than LPS group at T(3) and T(4). At T(6), the DO(2) and VO(2) were (9.5 +/- 1.3) and (4.1 +/- 1.5) ml/(kg.min), respectively, but there was no significant difference compared with LPS group. There was no significant difference in ERO(2) among groups (P > 0.05). CONCLUSION: The dexamethasone, aminoguanidin, amrinone can improve oxygen utilization in endotoxic shock rabbits, especially for dexamethasone and aminoguanidin. PMID- 14754536 TI - [Non-myeloperoxidase-mediated system activity of neutrophil in newborn infants]. AB - OBJECTIVE: To evaluate the variety of non-myeloperoxidase-mediated system activity of neutrophils in newborns during bacterial infection and the effect of cord plasma on the activation of non-myeloperoxidase-mediated system. METHODS: An infection model with Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) and a non-infection model with phorbol-12-myristate-13-acetate (PMA) were established to investigate the activation of non-myeloperoxidase-mediated system in neutrophils. According to the intensity of fluorescence, the activation of non myeloperoxidase-mediated system of neutrophils was detected by flow cytometry (FCM). The blood cells and plasma were separated from cord blood and adult blood and cross-mixed in order to investigate the opsonic activity. RESULTS: In the non infection model, the activation of non-myeloperoxidase-mediated system with PMA stimulation in cord blood was lower compared with that in adult blood, the statistical difference was significant (t = 3.378, P < 0.01). In the infection model, the activations of non-myeloperoxidase-mediated system in cord blood were also lower compared with those in adult blood, while the statistical difference could only be found in the model with E. coli stimulation (t = 12.150, P < 0.001). Furthermore the experiments demonstrated that cord plasma could deeply depress the non-myeloperoxidase-mediated system activity with E. coli stimulation. On the contrary, adult plasma could successfully recruit the potential of non-myeloperoxidase-mediated system activity of neutrophils in newborns. CONCLUSION: The function of neonatal neutrophils might not developed very well. As a stimulant, E. coli failed to induce the non-myeloperoxidase mediated system activity in neonates, which might be related to the lower level of immunoglobulins in cord blood. This result indicated that immunoglobulins played a more important modulating role in bacterial killing during gram-negative bacterial infections. PMID- 14754537 TI - [Anhidrotic ectodermal dysplasia: two cases in a family]. PMID- 14754538 TI - [Relationship between programmed cell death mechanisms and neuronal necrosis induced by seizures]. AB - OBJECTIVE: To clarify if programmed cell death mechanisms induced by seizures take part in the necrotic process of neurons. METHODS: Seizure was induced by pilocarpine (P) in Sprague-Dawley adult rats which were allowed to recover for 24 or 72 hours before perfusion-fixation. Neuronal death was assessed by light microscopy with the hematoxylin-eosin (HE) staining and with in situ terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL). Bax and Bcl-2 protein expression were examined by histochemistry. RESULTS: Twenty-four and 72 hours after seizures, neuronal death in hippocampus CA1 region was morphologically necrotic. TUNEL-positive and morphologically necrotic cells increased in the hippocampal CA1 region at 72 hours after seizures, there was significant difference compared with controls (P < 0.001). Bax expression was also increased in the hippocampal CA1 region at 72 hours after seizures (P < 0.001), but Bcl-2 expression did not increase, while Bcl-2/Bax ratio decreased. CONCLUSION: Seizures induced late-onset neuronal necrosis was accompanied by programmed cell death mechanisms. PMID- 14754539 TI - [Chronic Schonlein-Henock syndrome complicated with Helicobacter pylori infection in a case]. PMID- 14754540 TI - [Relationship between the serum anti-Mullerian hormone and testicular development]. PMID- 14754541 TI - [Correlation of acute respiratory tract infections with asthma exacerbation in children]. PMID- 14754542 TI - [Protective effect and mechanism of superoxide dismutase in CVB3m virus-induced cardiac muscle damage in mice]. PMID- 14754543 TI - [Expression of Ref-1 and FAP-1 mRNA in hypoxic-ischemic injury in neonatal rat]. PMID- 14754544 TI - [Effect of retinoic acid treatment on lung injury in hyperoxia-exposed newborn rats]. PMID- 14754545 TI - [Measurement of urinary lactate: creatine ratio for early identification of newborn infants at risk for the hypoxic-ischemic encephalopathy]. PMID- 14754546 TI - [Changes of insulin-like growth factor-II and insulin growth factor binding protein-3 in cerebrospinal fluid in children with central nervous system infection]. PMID- 14754547 TI - [Report of 24 cases with low back pain occurred after lumbar puncture]. PMID- 14754548 TI - [Acute respiratory distress syndrome in full term neonates, report of 4 cases]. PMID- 14754549 TI - [Analysis on misdiagnosis of inhaled organic phosphorus poisoning in 15 newborn infants]. PMID- 14754550 TI - [One case of hereditary tyrosinemia type I]. PMID- 14754551 TI - [Metatropic dwarfism in a case]. PMID- 14754552 TI - [Pulmonary embolism in children]. PMID- 14754553 TI - [Transforming growth factor beta and chronic lung disease of prematurity]. PMID- 14754554 TI - [Case No. 107--Abdominal distention, hematochezia and jaundice]. PMID- 14754555 TI - [Public health nursing in Greece]. AB - Nursing is considered, as the main profession that covers the need for primary health care services, in cooperation with physicians and other health and social care professions. In Greece, public or community health nursing as a concept exists since the beginning of professional nursing in the seconf half og the 19 century. There are three professions in Greece that are mainly involved in the provision of community nursing services. These are general nurses, health visitors, and midwives. General nurses are educated at degree level in the university or polytechnics. Health visitors and midwives are separate professions, also educated at degree level in polytechnics. Although all general nurses are prepared to work at any level of health care provision, they are not considered adequately educated to perform their full role in the community, with just their existing knowledge and experience. It is therefore essencial that further education or specialization is needed, in order that they could practice independently or as members of the primary health care teams effectively, and be able to respond to public health needs of the population PMID- 14754556 TI - [Communication difficulties between health professionals and foreign women in obstetrical and gynecological departments]. AB - Understanding difficulties discourage foreign patients to ask for health services. An epidemiological study has been carried out to evaluate language difficulties between them and health professionals. The article reports the results and discuss them. Results are important to be used to enhance and define different strategies to improve approaches and relations with foreign patients. PMID- 14754557 TI - [A proposal for programming at a didactic unit for nursing sciences (MED 45) in nursing degree courses]. AB - Pre-registration nursing education in Italy has been recently transferred in University. Therefore, many changes, evaluations and innovations have been implemented in Insubria University (Universita degli studi dell'Insubria, Varese). Starting from the reasons for the basis choice of three ample goals to reach in three years, a curricula proposal of the entire nursing disciplines and modules of the three years degree is presented. All nursing courses of the same disciplinary sectors ("MED 45" in the national university regulation) have been channelled into a few educational educational modules or units. The educational units of the first, second and third year are afterwards presented. The choice of these educational units is supported with examples while some tables show the topics of formal lectures together with the professionalising activities (i.e., laboratory, nursing skills acquisitiion, testimonies). Considering the difficulties one meets in evaluating teaching strategies, this three years experience has been positive for a few reasons. More precisely, it has contributed in decreasing the theory-practice gap, in developing critical thinking, and in answering to students'expectations at the beginning of the pre registration course. The introductory module has favoured the development of skilla and abilities useful for self-education and the students' future professional development. The lack of human and structural resources together with the sometimes negative influence of the clinical placements climate (reality shock) is among the main difficulties found. PMID- 14754558 TI - [Motivation, expectations, realization: from education to work]. AB - This paper presents the outcomes of a research oriented to exploration and description of the motivations and expectations that concur to assumption af an active and intentional role from students engagaged for achievement of knowledges, skills and highly qualitative consistent attitudes with practice of nursing service. PMID- 14754559 TI - [Oncology health professionals' perception regarding the problem of fatigue in cancer in Italy]. AB - Cancer relate fatigue is one of the most frequent problems among cancer patients. It is a nosological highly debilitating entity which has very negative effects on patients quality of life. Foreign literature reports that only during last years oncological operators have started getting aware and their training in order to face it is lacking. Our research wants to propose and analysis of the Italian reality by sending to oncological doctors and nurses a quationnaire. This questionnaire allowed to demonstrate that oncological operators have started knowing the fatigue problem but they are still unprepared about knowledge, professional capability and organizational modalities which could allow a better and efficacius approach to the fatigue problem. We think that research and professional training are the two ways which we will allow to gain knowledge and proper instruments in order to provide for our patients'needs. PMID- 14754560 TI - [Triage and heart disease at the emergency department]. AB - Our retrospective study of Nurses Triage schedules, has shown that the cardiac pathology represents one of the most frequent pathologies among those that arrive to the Niguarda Ca Granda Emergency Department. Furthermore, after an analysis of the schedulaes, we demostrated that the Triage system applied to the patients admitted to the Emergency Rooms with cardiac pathology was useful, safe and effectiveness, determining an early treatment of these patients. PMID- 14754561 TI - The effectiveness and cost-effectiveness of microwave and thermal balloon endometrial ablation for heavy menstrual bleeding: a systematic review and economic modelling. AB - OBJECTIVES: To estimate the clinical effectiveness and cost-effectiveness of microwave endometrial ablation (MEA) and thermal balloon endometrial ablation (TBEA) for heavy menstrual bleeding (HMB), compared with the existing (first generation) endometrial ablation (EA) techniques of transcervical resection (TCRE) and rollerball (RB) ablation, and hysterectomy. DATA SOURCES: Electronic databases, bibliographies of articles, and also experts in the field and relevant industry bodies were asked to provide information. REVIEW METHODS: A detailed search strategy was carried out to identify systematic reviews and controlled trials of MEA and TBEA versus first-generation techniques for EA. In addition to electronic database searching, reference lists were hand-searched and information sought from manufacturers of EA devices and by experts in the field. A deterministic Markov model was developed to assess cost-effectiveness. Data for the model were taken from a range of sources. RESULTS: The systematic review of first-generation EA techniques versus hysterectomy found that EA offered an alternative to hysterectomy for HMB, with fewer complications and a shorter recovery period. Satisfaction and effectiveness were high for both MEA and TBEA. Costs were lower with EA although the difference narrows over time. Second generation EA techniques are an alternative treatment to first-generation techniques for HMB, and first-generation techniques are known to offer an alternative to hysterectomy. Although no trials of second-generation techniques and hysterectomy have been undertaken, it seems reasonable to assume that second generation techniques also offer an alternative surgical treatment. Using the model to assess cost-effectiveness, costs were very slightly higher for MEA when compared to TBEA, and differences in quality-adjusted life-years (QALYs) were negligible. For MEA compared with transcervical resection of the endometrium (TCRE) and RB ablation, costs were slightly lower with MEA and MEA accrued very slightly more QALYs. Compared with hysterectomy, MEA costs less and accrues slightly fewer QALYs. For TBEA compared with TCRE and RB ablation, costs were lower with TBEA and TBEA accrued slightly more QALYs. Compared with hysterectomy, TBEA costs moderately less and accrues moderately fewer QALYs. CONCLUSIONS: Overall, there were few significant differences between the outcomes of first- and second-generation techniques including bleeding, satisfaction and QoL measures and repeat surgery rates. Second-generation techniques had significantly shorter operating and theatre times and there appear to be fewer serious perioperative adverse effects with second-generation techniques and postoperative effects are similar. Compared with hysterectomy, TCRE and RB are quicker to perform and result in shorter hospitalisation and faster return to work. Hysterectomy results in more adverse effects and is more expensive, although the need for retreatment leads this difference to decrease over time. Satisfaction with hysterectomy is initially higher, but there is no significant difference after 2 years. The economic model suggests that second-generation techniques are more cost-effective than first-generation techniques of EA for HMB. Both TBEA and MEA appear to be less costly than hysterectomy, although the latter results in more QALYs. Further research is suggested to make direct comparisons of the cost effectiveness of second-generation EA techniques, to carry out longer term follow up for all methods of EA in RCTs, and to develop more sophisticated modelling studies. Further research is also recommended into HMB to establish health-state utility values, its surgical treatment, convalescence, complications of treatment, symptoms and patient satisfaction. PMID- 14754564 TI - Is public support for environmental protection decreasing? An analysis of U.S. and New Jersey data. AB - Telephone surveys made of 800-1,000 randomly selected residents of the United States and New Jersey in 2003 show a sharp decline in support for antipollution regulations, although pollution remains a major concern. This drop in support is associated with slowing of the economy, fear of terrorism, and other competing priorities. The leading proponents of maintaining strong environmental regulations are relatively affluent mainstream white Americans. Despite this recent drop in support, overt attempts to weaken the basic regulations are likely to face stiff opposition unless there is an obvious economic downturn or increasing terrorism that causes a larger proportion of the public to feel that weakening environmental regulations will increase jobs and security. Key words: age, environmental laws and regulations, perception, polls, public support, race/ethnicity, trends. PMID- 14754562 TI - Systematic review and modelling of the investigation of acute and chronic chest pain presenting in primary care. AB - OBJECTIVES: To ascertain the value of a range of methods - including clinical features, resting and exercise electrocardiography, and rapid access chest pain clinics (RACPCs) - used in the diagnosis and early management of acute coronary syndrome (ACS), suspected acute myocardial infarction (MI), and exertional angina. DATA SOURCES: MEDLINE, EMBASE, CINAHL, the Cochrane Library and electronic abstracts of recent cardiological conferences. REVIEW METHODS: Searches identified studies that considered patients with acute chest pain with data on the diagnostic value of clinical features or an electrocardiogram (ECG); patients with chronic chest pain with data on the diagnostic value of resting or exercise ECG or the effect of a RACPC. Likelihood ratios (LRs) were calculated for each study, and pooled LRs were generated with 95% confidence intervals. A Monte Carlo simulation was performed evaluating different assessment strategies for suspected ACS, and a discrete event simulation evaluated models for the assessment of suspected exertional angina. RESULTS: For acute chest pain, no clinical features in isolation were useful in ruling in or excluding an ACS, although the most helpful clinical features were pleuritic pain (LR+ 0.19) and pain on palpation (LR+ 0.23). ST elevation was the most effective ECG feature for determining MI (with LR+ 13.1) and a completely normal ECG was reasonably useful at ruling this out (LR+ 0.14). Results from 'black box' studies of clinical interpretation of ECGs found very high specificity, but low sensitivity. In the simulation exercise of management strategies for suspected ACS, the point of care testing with troponins was cost-effective. Pre-hospital thrombolysis on the basis of ambulance telemetry was more effective but more costly than if performed in hospital. In cases of chronic chest pain, resting ECG features were not found to be very useful (presence of Q-waves had LR+ 2.56). For an exercise ECG, ST depression performed only moderately well (LR+ 2.79 for a 1 mm cutoff), although this did improve for a 2 mm cutoff (LR+ 3.85). Other methods of interpreting the exercise ECG did not result in dramatic improvements in these results. Weak evidence was found to suggest that RACPCs may be associated with reduced admission to hospital of patients with non-cardiac pain, better recognition of ACS, earlier specialist assessment of exertional angina and earlier diagnosis of non-cardiac chest pain. In a simulation exercise of models of care for investigation of suspected exertional angina, RACPCs were predicted to result in earlier diagnosis of both confirmed coronary heart disease (CHD) and non-cardiac chest pain than models of care based around open access exercise tests or routine cardiology outpatients, but they were more expensive. The benefits of RACPCs disappeared if waiting times for further investigation (e.g. angiography) were long (6 months). CONCLUSIONS: Where an ACS is suspected, emergency referral is justified. ECG interpretation in acute chest pain can be highly specific for diagnosing MI. Point of care testing with troponins is cost-effective in the triaging of patients with suspected ACS. Resting ECG and exercise ECG are of only limited value in the diagnosis of CHD. The potential advantages of RACPCs are lost if there are long waiting times for further investigation. Recommendations for further research include the following: determining the most appropriate model of care to ensure accurate triaging of patients with suspected ACS; establishing the cost-effectiveness of pre-hospital thrombolysis in rural areas; determining the relative cost-effectiveness of rapid access chest pain clinics compared with other innovative models of care; investigating how rapid access chest pain clinics should be managed; and establishing the long-term outcome of patients discharged from RACPCs. PMID- 14754565 TI - A high-level disinfection standard for land applying sewage sludges (biosolids). AB - Complaints associated with land-applied sewage sludges primarily involve irritation of the skin, mucous membranes, and the respiratory tract accompanied by opportunistic infections. Volatile emissions and organic dusts appear to be the main source of irritation. Occasionally, chronic gastrointestinal problems are reported by affected residents who have private wells. To prevent acute health effects, we recommend that the current system of classifying sludges based on indicator pathogen levels (Class A and Class B) be replaced with a single high level disinfection standard and that methods used to treat sludges be improved to reduce levels of irritant chemicals, especially endotoxins. A national opinion survey of individuals impacted by or concerned about the safety of land application practices indicated that most did not consider the practice inherently unsafe but that they lacked confidence in research supported by federal and state agencies. PMID- 14754566 TI - An evaluation of the environmental and health effects of vehicle exhaust catalysts in the UK. AB - Since 1993, all new gasoline-engine automobiles in the United Kingdom have been supplied with three-way vehicle exhaust catalytic converters (VECs) containing platinum, palladium, and rhodium, to comply with European Commission Stage I limits on emissions of regulated pollutants: carbon monoxide, hydrocarbons, and oxides of nitrogen. We conducted a physical and economic evaluation of the environmental and health benefits from a reduction in emissions through this mandated environmental technology against the costs, with reference to urban areas in Great Britain. We made both an ex post assessment--based on available data to 1998--and an ex ante assessment--projected to 2005, the year when full penetration of VECs into the fleet is expected. Substantial health benefits in excess of the costs of VECs were indicated: By 1998 the estimated net societal health benefits were approximately 500 million British pounds, and by 2005 they were estimated to rise to as much as 2 billion British pounds. We also found through environmental surveys that although lead in road dust has fallen by 50% in urban areas, platinum accumulations near roads have risen significantly, up to 90-fold higher than natural background levels. This rapid accumulation of platinum suggests further monitoring is warranted, although as yet there is no evidence of adverse health effects. PMID- 14754567 TI - Agricultural task and exposure to organophosphate pesticides among farmworkers. AB - Little is known about pesticide exposure among farmworkers, and even less is known about the exposure associated with performing specific farm tasks. Using a random sample of 213 farmworkers in 24 communities and labor camps in eastern Washington State, we examined the association between occupational task and organophosphate (OP) pesticide residues in dust and OP metabolite concentrations in urine samples of adult farmworkers and their children. The data are from a larger study that sought to test a culturally appropriate intervention to break the take-home pathway of pesticide exposure. Commonly reported farm tasks were harvesting or picking (79.2%), thinning (64.2%), loading plants or produce (42.2%), planting or transplanting (37.6%), and pruning (37.2%). Mixing, loading, or applying pesticide formulations was reported by 20% of our sample. Workers who thinned were more likely than those who did not to have detectable levels of azinphos-methyl in their house dust (92.1% vs. 72.7%; p = 0.001) and vehicle dust (92.6% vs. 76.5%; p = 0.002). Thinning was associated with higher urinary pesticide metabolite concentrations in children (91.9% detectable vs. 81.3%; p = 0.02) but not in adults. Contrary to expectation, workers who reported mixing, loading, or applying pesticide formulations had lower detectable levels of pesticide residues in their house or vehicle dust, compared with those who did not perform these job tasks, though the differences were not significant. Future research should evaluate workplace protective practices of fieldworkers and the adequacy of reentry intervals for pesticides used during thinning. PMID- 14754568 TI - Developmental exposure to chlorpyrifos elicits sex-selective alterations of serotonergic synaptic function in adulthood: critical periods and regional selectivity for effects on the serotonin transporter, receptor subtypes, and cell signaling. AB - During brain development, serotonin (5HT) provides essential neurotrophic signals, and in earlier work, we found that developmental exposure to chlorpyrifos (CPF) elicits short-term changes in 5HT systems. In the present study, we evaluated the effects in adulthood after CPF exposures from the neural tube stage [gestational days (GD) 9-12] and the late gestational period (GD17-20) through postnatal neuronal differentiation and synaptogenesis [postnatal days (PN) 1-4 and 11-14], using treatments below the threshold for systemic toxicity. With exposure on GD9-12, CPF elicited global elevations in 5HT1A and 5HT2 receptors and in the 5HT presynaptic transporter. The GD17-20 treatment elicited larger effects that displayed selectivity for regions with 5HT nerve terminals and that were preferential for males. Although similar receptor up-regulation was seen after PN1-4 exposure, the effects were larger in regions with 5HT cell bodies; in addition, the presynaptic transporter was down-regulated in the nerve terminal zones of females. The PN11-14 exposure had much smaller effects on receptors but still elicited transporter suppression with the same regional and sex selectivity. Although CPF exposure on GD17-20, PN1-4, or PN11-14 altered the ability of 5HT to modulate adenylyl cyclase, this change did not correspond with the effects on 5HT receptors, suggesting an additional set of effects on proteins that transduce the 5HT signal. Our results indicate that CPF elicits long-lasting changes in 5HT receptors, the presynaptic 5HT transporter, and 5HT-mediated signal transduction after exposure in discrete developmental windows that range from the neural tube stage through synaptogenesis. These effects are likely to contribute to neurobehavioral teratology of CPF. PMID- 14754569 TI - Potential exposure to PCBs, DDT, and PBDEs from sport-caught fish consumption in relation to breast cancer risk in Wisconsin. AB - In Wisconsin, consumption of Great Lakes fish is an important source of exposure to polychlorinated biphenyls (PCBs), dichlorodiphenyltrichloroethane (DDT), polybrominated diphenyl ethers (PBDEs), and other halogenated hydrocarbons, all of which may act as potential risk factors for breast cancer. We examined the association between sport-caught fish consumption and breast cancer incidence as part of an ongoing population-based case-control study. We identified breast cancer cases 20-69 years of age who were diagnosed in 1998-2000 (n = 1,481) from the Wisconsin Cancer Reporting System. Female controls of similar age were randomly selected from population lists (n = 1,301). Information about all sport caught (Great Lakes and other lakes) fish consumption and breast cancer risk factors was obtained through telephone interviews. After adjustment for known and suspected risk factors, the relative risk of breast cancer for women who had recently consumed sport-caught fish was similar to women who had never eaten sport-caught fish [relative risk (RR) = 1.00; 95% confidence interval (CI), 0.86 1.17]. Frequency of consumption and location of sport-caught fish were not associated with an increased risk of breast cancer. Recent consumption of Great Lakes fish was not associated with postmenopausal breast cancer (RR = 0.78; 95% CI, 0.57-1.07), whereas risk associated with premenopausal breast cancer was elevated (RR = 1.70; 95% CI, 1.16-2.50). In this study we found no overall association between recent consumption of sport-caught fish and breast cancer, although there may be an increased breast cancer risk for subgroups of women who are young and/or premenopausal. PMID- 14754570 TI - Highly chlorinated PCBs inhibit the human xenobiotic response mediated by the steroid and xenobiotic receptor (SXR). AB - Polychlorinated biphenyls (PCBs) are a family of persistent organic contaminants suspected to cause adverse effects in wildlife and humans. In rodents, PCBs bind to the aryl hydrocarbon (AhR) and pregnane X receptors (PXR) inducing the expression of catabolic cytochrome p450 enzymes of the CYP1A and 3A families. We found that certain highly chlorinated PCBs are potent activators of rodent PXR but antagonize its human ortholog, the steroid and xenobiotic receptor (SXR), inhibiting target gene induction. Thus, exposure to PCBs may blunt the human xenobiotic response, inhibiting the detoxification of steroids, bioactive dietary compounds, and xenobiotics normally mediated by SXR. The antagonistic PCBs are among the most stable and abundant in human tissues. These findings have important implications for understanding the biologic effects of PCB exposure and the use of animal models to predict the attendant risk. PMID- 14754571 TI - Developmental effects of chlorpyrifos extend beyond neurotoxicity: critical periods for immediate and delayed-onset effects on cardiac and hepatic cell signaling. AB - The fetal and neonatal neurotoxicity of chlorpyrifos (CPF) and related insecticides is a major concern. Developmental effects of CPF involve mechanisms over and above cholinesterase inhibition, notably events in cell signaling that are shared by nonneural targets. In the present study, we evaluated the immediate and long-term effects of CPF exposure of rats during different developmental windows [gestational days (GD) 9-12 or 17-20, postnatal days (PN) 1-4 or 11-14] on the adenylyl cyclase (AC) signaling cascade in the heart and liver. In addition to basal AC activity, we assessed the responses to direct AC stimulants (forskolin, Mn2+); to isoproterenol and glucagon, which activate signaling through specific membrane receptors; and to sodium fluoride, which activates the G-proteins that couple the receptors to AC. Few immediate effects on AC were apparent when CPF doses remained below the threshold for systemic toxicity. Nevertheless, CPF exposures on GD9-12, GD17-20, or PN1-4 elicited sex-selective effects that emerged by adulthood (PN60), whereas later exposure (PN11-14) elicited smaller, nonsignificant effects, indicative of closure of the window of vulnerability. Most of the effects were heterologous, involving signaling elements downstream from the receptors, and thus were shared by multiple inputs; superimposed on this basic pattern, there were also selective alterations in receptor-mediated responses. These results suggest that the developmental toxicity of CPF extends beyond the nervous system, to include cell signaling cascades that are vital to cardiac and hepatic homeostasis. Future work needs to address the potential implications of these effects for cardiovascular and metabolic disorders that may emerge long after the end of CPF exposure. PMID- 14754572 TI - Cause-specific mortality and the extended effects of particulate pollution and temperature exposure. AB - Air pollution exposure studies in the past decade have focused on acute (days) or long-term (years) effects. We present an analysis of medium-term (weeks to months) exposure effects of particulate pollution and temperature. We assessed the associations of particulate pollution (black smoke) and temperature with age standardized daily mortality rates over 17 years in Dublin, Ireland, using a polynomial distributed lag model of both temperature and particulate air pollution simultaneously through 40 days after exposure. When only acute effects (3-day mean) were considered, we found total mortality increased by 0.4% for each 10-microg/m3 increase in black smoke concentration. When deaths in the 40 days after exposure were considered, we found a 1.1% increase. For respiratory mortality, the estimated effect was 0.9% for acute exposures, but 3.6% for the extended follow-up. We found each increase in current-day temperature by 1 degree C was associated with a 0.4% increase in total mortality, whereas each decrease of 1 degree C was associated with a 2.6% increase in mortality in the following 40 days. For both temperature and pollution, the largest effects on cardiovascular mortality were observed immediately, whereas respiratory mortality was delayed and distributed over several weeks. These effects were two to three times greater than the acute effects reported in other studies, and approach the effects reported in longer-term survival studies. This analysis suggests that studies on the acute effects of air pollution have underestimated the total effects of temperature and particulate air pollution on mortality. PMID- 14754573 TI - Concentrations of dialkyl phosphate metabolites of organophosphorus pesticides in the U.S. population. AB - We report population-based concentrations, stratified by age, sex, and racial/ethnic groups, of dialkyl phosphate (DAP) metabolites of multiple organophosphorus pesticides. We measured dimethylphosphate (DMP), dimethylthiophosphate (DMTP), dimethyldithiophosphate (DMDTP), diethylphosphate (DEP), diethylthiophosphate (DETP), and diethyldithiophosphate (DEDTP) concentrations in 1,949 urine samples collected in U.S. residents 6-59 years of age during 1999 and 2000 as a part of the ongoing National Health and Nutrition Examination Survey (NHANES). We detected each DAP metabolite in more than 50% of the samples, with DEP being detected most frequently (71%) at a limit of detection of 0.2 microg/L. The geometric means for the metabolites detected in more than 60% of the samples were 1.85 microg/L for DMTP and 1.04 microg/L for DEP. The 95th percentiles for each metabolite were DMP, 13 microg/L; DMTP, 46 microg/L; DMDTP, 19 micro g/L; DEP, 13 microg/L; DETP, 2.2 microg/L; and DEDTP, 0.87 microg/L. We determined the molar sums of the dimethyl-containing and diethyl-containing metabolites; their geometric mean concentrations were 49.4 and 10.5 nmol/L, respectively, and their 95th percentiles were 583 and 108 nmol/L, respectively. These data are also presented as creatinine-adjusted concentrations. Multivariate analyses showed concentrations of DAPs in children 6 11 years of age that were consistently significantly higher than in adults and often higher than in adolescents. Although the concentrations between sexes and among racial/ethnic groups varied, no significant differences were observed. These data will be important in evaluating the impact of organophosphorus pesticide exposure in the U.S. population and the effectiveness of regulatory actions. PMID- 14754574 TI - Vanadyl sulfate inhibits NO production via threonine phosphorylation of eNOS. AB - Exposure to excessive vanadium occurs in some occupations and with consumption of some dietary regimens for weight reduction and body building. Because vanadium is vasoactive, individuals exposed to excessive vanadium may develop adverse vascular effects. We have previously shown that vanadyl sulfate causes acute pulmonary vasoconstriction, which could be attributed in part to inhibition of nitric oxide production. In the present study we investigated whether NO inhibition was related to phosphorylation of endothelial nitric oxide synthase (eNOS). VOSO4 produced dose-dependent constriction of pulmonary arteries in isolated perfused lungs and pulmonary arterial rings and a right shift of the acetylcholine-dependent vasorelaxation curve. VOSO4 inhibited constitutive as well as A23187-stimulated NO production. Constitutive NO inhibition was accompanied by increased Thr495 (threonine at codon 495) phosphorylation of eNOS, which would inhibit eNOS activity. Thr495 phosphorylation of eNOS and inhibition of NO were partially reversed by pretreatment with calphostin C, a protein kinase C (PKC) inhibitor. There were no changes in Ser1177 (serine at codon 1177) or tyrosine phosphorylation of eNOS. These results indicate that VOSO4 induced acute pulmonary vasoconstriction that was mediated in part by the inhibition of endothelial NO production via PKC-dependent phosphorylation of Thr495 of eNOS. Exposure to excessive vanadium may contribute to pulmonary vascular diseases. PMID- 14754575 TI - Dichlorodiphenyldichloroethane burden and breast cancer risk: a meta-analysis of the epidemiologic evidence. AB - The relationship of dichlorodiphenyltrichloroethane (DDT) exposure and breast cancer risk has received increasing attention since the beginning of the 1990s. Contradicting published results regarding the relationship between body burden levels of p,p'-dichlorodiphenyldichloroethane (p,p'-DDE)--the main DDT metabolite -and breast cancer, we argue that such differences stem from methodologic differences among those studies. We performed a meta-analysis of 22 articles using DerSimonian and Laird's method for random effects models. The Q-statistic was used to identify heterogeneity in the outcome variable across studies. The gradient of p,p'-DDE exposure in epidemiologic studies was homogenized to serum lipid bases (nanograms per gram). The potential for publication bias was examined by means of the Begg's test. We discuss methodologic features of the studies in an attempt to reconcile the findings. The summary odds ratio (OR) for selected studies was 0.97 (95% confidence interval, 0.87-1.09) and the gradient of exposure ranged from 84.37 to 12,948 ng/g. No overall heterogeneity in the OR was observed (chi-squared = 27.93; df = 23; p = 0.218). Neither the study design nor the lack of breast-feeding control or the type of biologic specimen used to measure p,p'-DDE levels were the causes of heterogeneity throughout the studies. Evidence for publication bias was not found (p = 0.253). Overall, these results should be regarded as a strong evidence to discard the putative relationship between p,p'-DDE and breast cancer risk. Nevertheless, the exposure to DDT during critical periods of human development--from conception to adolescence--and individual variations in metabolizing enzymes of DDT or its derivatives are still important areas to be researched in regard to breast cancer development in adulthood. PMID- 14754576 TI - Assessing potential risk of heavy metal exposure from consumption of home produced vegetables by urban populations. AB - We performed a risk assessment of metal exposure to population subgroups living on, and growing food on, urban sites. We modeled uptake of cadmium, copper, nickel, lead, and zinc for a selection of commonly grown allotment and garden vegetables. Generalized linear cross-validation showed that final predictions of Cd, Cu, Ni, and Zn content of food crops were satisfactory, whereas the Pb uptake models were less robust. We used predicted concentrations of metals in the vegetables to assess the risk of exposure to human populations from homegrown food sources. Risks from other exposure pathways (consumption of commercially produced foodstuffs, dust inhalation, and soil ingestion) were also estimated. These models were applied to a geochemical database of an urban conurbation in the West Midlands, United Kingdom. Risk, defined as a "hazard index," was mapped for three population subgroups: average person, highly exposed person, and the highly exposed infant (assumed to be a 2-year-old child). The results showed that food grown on 92% of the urban area presented minimal risk to the average person subgroup. However, more vulnerable population subgroups (highly exposed person and the highly exposed infant) were subject to hazard index values greater than unity. This study highlights the importance of site-specific risk assessment and the "suitable for use" approach to urban redevelopment. PMID- 14754577 TI - An unexplained case of elevated blood lead in a Hispanic child. AB - A 6-month-old child presented to a local pediatrician with an elevated blood lead level (BLL) of 41 microg/dL. The child was treated as an outpatient for chelation therapy by a toxicologist. Subsequent BLLs obtained at 8 and 13 months of age were 40 microg/dL and 42 microg/dL, respectively. Siblings and family members had BLLs < 5 microg/dL except for the mother, who had a BLL of 14 microg/dL when the child was 6 months of age. Home inspections and phone calls to the family revealed no sources of lead from paint, dust, toys, mini-blinds, keys, food, water, or any take-home exposure. The family denied use of folk remedies such as Greta and Azarcon. The child was breast-fed, but the mother's BLL was not sufficiently high to explain the elevated BLL in the child. Housekeeping was excellent. The mother did admit to cooking beans in Mexican pottery (pieces found outside were positive for lead), but she discontinued use after the initial lead check at 6 months. The bean pot was not a likely source, as none of the family had elevated BLLs including a 5-year-old sister. Follow-up testing of blood lead when the child was 15 months of age revealed values of 28 microg/dL for the child and 9 microg/dL for the mother. Subsequent testing of the child shows a slow decline. The slow release of lead suggests depletion of bone stores acquired during pregnancy, possibly due to pica behavior of the mother during pregnancy. PMID- 14754578 TI - Assessing the health benefits of air pollution reduction for children. AB - Benefit-cost analyses of environmental regulations are increasingly mandated in the United States. Evaluations of criteria air pollutants have focused on benefits and costs associated with adverse health effects. Children are significantly affected by the health benefits of improved air quality, yet key environmental health policy analyses have not previously focused specifically on children's effects. In this article we present a "meta-analysis" approach to child-specific health impacts derived from the U.S. Clean Air Act (CAA). On the basis of data from existing studies, reductions in criteria air pollutants predicted to occur by 2010 because of CAA regulations are estimated to produce the following impacts: 200 fewer expected cases of postneonatal mortality; 10,000 fewer asthma hospitalizations in children 1-16 years old, with estimated benefits ranging from 20 million U.S. dollars to 46 million U.S. dollars (1990 U.S. dollars); 40,000 fewer emergency department visits in children 1-16 years old, with estimated benefits ranging from 1.3 million U.S. dollars to 5.8 million U.S. dollars; 20 million school absences avoided by children 6-11 years old, with estimated benefits of 0.7-1.8 billion U.S. dollars; and 10,000 fewer infants of low birth weight, with estimated benefits of 230 million U.S. dollars. Inclusion of limited child-specific data on hospitalizations, emergency department visits, school absences, and low birth weight could be expected to add 1-2 billion U.S. dollars (1990 U.S. dollars) to the 8 billion U.S. dollars in health benefits currently estimated to result from decreased morbidity, and 600 million U.S. dollars to the 100 billion U.S. dollars estimated to result from decreased mortality. These estimates highlight the need for increased consideration of children's health effects. Key needs for environmental health policy analyses include improved information for children's health effects, additional life-stage specific information, and improved health economics information specific for children. PMID- 14754579 TI - Effect of succimer on growth of preschool children with moderate blood lead levels. AB - Growth deficits associated with lead exposure might be ameliorated by chelation. We examined the effect of succimer on growth in 780 children 12-33 months old who had blood lead levels of 20-44 microg/dL and were randomized to receive up to three 26-day courses of succimer or placebo in a multicenter, double-blind trial. The difference in changes in weight and height between succimer and placebo groups at 1-34 months was calculated by fitting cubic splines. The difference in height change in children on succimer compared with placebo was -0.27 cm [95% confidence interval (95% CI), -0.42 to -0.11] from baseline to 9 months, when 99% of children had completed treatment, and -0.43 cm (95% CI, -0.77 to -0.09) during 34 months of follow-up. Similar differences in weight gain were not statistically significant. Although succimer lowers blood lead in moderately lead-poisoned children, it does not have a beneficial effect on growth and may have an adverse effect. PMID- 14754580 TI - A framework for assessing risks to children from exposure to environmental agents. AB - In recent years there has been an increasing focus in environmental risk assessment on children as a potentially susceptible population. There also has been growing recognition of the need for a systematic approach for organizing, evaluating, and incorporating the available data on children's susceptibilities in risk assessments. In this article we present a conceptual framework for assessing risks to children from environmental exposures. The proposed framework builds on the problem formulation-->analysis-->risk characterization paradigm, identifying at each phase the questions and issues of particular importance for characterizing risks to the developing organism (from conception through organ maturation). The framework is presented and discussed from the complementary perspectives of toxicokinetics and toxicodynamics. PMID- 14754582 TI - Hazard identification and predictability of children's health risk from animal data. AB - Children differ from adults both physiologically and behaviorally. These differences can affect how and when exposures to xenobiotics occur and the resulting responses. Testing using animal models may be used to predict whether children display novel toxicities not observed in adults or whether children are more or less sensitive to known toxicities. Historically, evaluation of developmental toxicity has focused on gestational exposures and morphological changes resulting from this exposure. Functional consequences of gestational exposure and postnatal exposure have not been as well studied. Difficulties with postnatal toxicity evaluations include divergent differentiation of structure, function and physiology across species, lack of understanding of species differences in functional ontogeny, and lack of common end points and milestones across species. PMID- 14754581 TI - Children's health and the environment: public health issues and challenges for risk assessment. AB - Infants and children are not little adults. They are uniquely vulnerable to environmental toxicants. To protect infants and children against toxicants, the National Research Council in 1993 called for development of an approach to risk assessment that considers children's unique patterns of exposure and their special vulnerabilities to pesticides. Many aspects of that call were codified into federal law in the Food Quality Protection Act (FQPA) of 1996. This report highlights the central elements needed for development of a child-protective approach to risk assessment: a) improved quantitative assessment of children's exposures at different life stages, from fetal life through adolescence, including acute and chronic exposures, exposures via multiple routes, and exposures to multiple agents; b) development of new approaches to toxicity testing of chemicals that can detect unanticipated and subtle outcomes and that evaluate experimental subjects over the entire life span from early exposure to natural death to replicate the human experience; c) development of new toxicodynamic and toxicokinetic models that account for the unique physiologic characteristics of infants and children; d) development of new approaches to assessment of outcomes, functional, organ, cellular and molecular, over the entire life span; these measures need to be incorporated into toxicity testing and into long-term prospective epidemiologic studies of children; and e) application of uncertainty and safety factors in risk assessment that specifically consider children's risks. Under FQPA, children are presumed more vulnerable to pesticides than adults unless evidence exists to the contrary. Uncertainty and safety factors that are protective of children must therefore be incorporated into risk assessment when data on developmental toxicity are lacking or when there is evidence of developmental toxicity. The adequate protection of children against toxic agents in the environment will require fundamental and far reaching revisions of current approaches to toxicity testing and risk assessment. PMID- 14754584 TI - The growth and control of international environmental crime. PMID- 14754583 TI - Incorporating children's toxicokinetics into a risk framework. AB - Children's responses to environmental toxicants will be affected by the way in which their systems absorb, distribute, metabolize, and excrete chemicals. These toxicokinetic factors vary during development, from in utero where maternal and placental processes play a large role, to the neonate in which emerging metabolism and clearance pathways are key determinants. Toxicokinetic differences between neonates and adults lead to the potential for internal dosimetry differences and increased or decreased risk, depending on the mechanisms for toxicity and clearance of a given chemical. This article raises a number of questions that need to be addressed when conducting a toxicokinetic analysis of in utero or childhood exposures. These questions are organized into a proposed framework for conducting the assessment that involves problem formulation (identification of early life stage toxicokinetic factors and chemical-specific factors that may raise questions/concerns for children); data analysis (development of analytic approach, construction of child/adult or child/animal dosimetry comparisons); and risk characterization (evaluation of how children's toxicokinetic analysis can be used to decrease uncertainties in the risk assessment). The proposed approach provides a range of analytical options, from qualitative to quantitative, for assessing children's dosimetry. Further, it provides background information on a variety of toxicokinetic factors that can vary as a function of developmental stage. For example, the ontology of metabolizing systems is described via reference to pediatric studies involving therapeutic drugs and evidence from in vitro enzyme studies. This type of resource information is intended to help the assessor begin to address the issues raised in this paper. PMID- 14754585 TI - Radiographic abnormalities and asbestos exposure: Libby, Montana. PMID- 14754588 TI - Overstating the consequences of radiographic abnormalities. PMID- 14754589 TI - Expert witnesses need to know about the new risks. PMID- 14754590 TI - Validation of (Q)SARs models. PMID- 14754594 TI - Environmental crimes: profiting at earth's expense. PMID- 14754595 TI - Does secrecy equal security? Limiting access to environmental information. PMID- 14754596 TI - A cleaner doctor's kit: tiny bubbles mean huge improvements. PMID- 14754598 TI - Clonal nature of hematopoietic stem cell disorders. PMID- 14754599 TI - Conventional and novel tools for defining the risk of the individual patient with chronic myeloid leukemia and for monitoring treatment. PMID- 14754600 TI - Chronic lymphocytic leukemia 2003. PMID- 14754601 TI - Fanconi's anemia cell lines show distinct mechanisms of cell death in response to mitomycin C or agonistic anti-Fas antibodies. AB - BACKGROUND AND OBJECTIVES: Fanconi anemia (FA) cells are characteristically hypersensitive to bifunctional alkylating agents, notably mitomycin C (MMC), causing increased programmed cell death (PCD). FA cells also have abnormalities in mitochondrial function. We hypothesized that the abnormalities in PCD are mitochondrially mediated. We examined mitochondrial function in FA cells, comparing the intrinsic death pathway induced by MMC with the extrinsic pathway via Fas antibody, which can bypass the mitochondria. DESIGN AND METHODS: Normal and FA lymphoblastoid cell lines were treated with MMC or agonistic anti-Fas antibody. PCD was assessed using flow cytometry, Western blot analysis, and DNA gel electrophoresis. RESULTS: FA cells showed hypersensitivity to MMC, but slight resistance to Fas-mediated PCD. MMC induced chromatin condensation, but not apoptotic body formation. Fas induced classical apoptosis. MMC failed to induce mitochondrial depolarization, while some depolarization occurred with anti-Fas. These results suggested that MMC failed to induce caspase activity in FA cells. No cleavage of caspase 3 was observable and PCD was not inhibited by the caspase inhibitor zVAD-fmk. Fas-induced caspase 3 cleavage, and cell death was inhibited by zVAD-fmk. There were common downstream abnormalities in the execution phase of PCD, as both agonists failed to cleave PARP, or to induce nucleosomal fragmentation. INTERPRETATION AND CONCLUSIONS: Our results suggest that mitochondrial function in FA cells is abnormal, resulting in necrotic or caspase independent PCD, but that further abnormalities may exist downstream of the mitochondria. This may have implications in explaining in vivo aspects of FA. PMID- 14754603 TI - Clinico-biological features and outcome of acute promyelocytic leukemia patients with persistent polymerase chain reaction-detectable disease after the AIDA front line induction and consolidation therapy. AB - BACKGROUND AND OBJECTIVES: Front line treatment of acute promyelocytic leukemia (APL) with all-trans retinoic acid (ATRA) and chemotherapy (CHT) results in molecular remission in approximately 95% of patients tested after consolidation. The small fraction of patients with persistence of molecular disease (i.e. those in whom polymerase chain reaction (PCR) is positive for PML/RARalpha) after such therapy are thought to have a dismal prognosis but has not yet been investigated in detail. DESIGN AND METHODS: We analyzed the clinico-biological features at presentation of APL patients who showed PCR-detectable residual disease and compared them to those of patients achieving molecular remission after AIDA induction and consolidation. Furthermore, we report the outcome of patients with molecularly persistent disease treated with salvage therapy. RESULTS: Patients attaining molecular remission (n=650) and patients who tested PCR+ve at the end of consolidation (n=23) were not statistically significantly different as regards median age, white cell and platelet counts, morphologic subtype (M3 or M3v), fibrinogen levels or PML/RARalpha transcript type. As to treatment outcome after salvage therapy, 7 patients were treated before morphologic relapse [3 with chemotherapy and autologous stem cell transplantation (SCT) and 4 with allogeneic SCT], and are alive after 64-118 months. Of 16 patients treated at the time of morphologic relapse, only 2 patients are alive, both of whom received an allogeneic SCT. INTERPRETATION AND CONCLUSIONS: Our findings indicate that APL patients who are molecularly resistant to the AIDA protocol have no distinguishing features at presentation. Their outcome suggests the need for early therapeutic intervention with aggressive treatment prior to the occurrence of hematologic relapse. PMID- 14754602 TI - Primitive hematopoietic stem cells shows a polyclonal pattern in myelodysplastic syndromes. AB - BACKGROUND AND OBJECTIVES: Clonal hematopoiesis is the hallmark of myelodysplastic syndromes, but the role played by pluripotent stem cells and progenitor cells in these disorders remains unclear. DESIGN AND METHODS: Eight female patients with myelodysplastic syndrome were studied. X-chromosome inactivation patterns were analyzed in peripheral blood granulocytes, T lymphocytes, single colonies originating from bone marrow progenitors and pluripotent stem cells, using the human androgen receptor locus polymorphism assay. RESULTS: Granulocytes and progenitor cells were monoclonal in 7/8 cases. Immature stem cells showed a non-clonal pattern of X-inactivation and were detectable at diagnosis in the presence of clonal hematopoiesis. T-lymphocyte clonality was heterogeneous. INTERPRETATION AND CONCLUSIONS: In myelodysplastic syndromes, hematopoiesis may be dominated by a neoplastic clone by virtue of its biological advantage over a residual polyclonal, probably still normal, population of immature stem cells still able to grow in vitro. PMID- 14754604 TI - P-glycoprotein and multidrug resistance associated protein-1 activity in 132 acute myeloid leukemias according to FAB subtypes and cytogenetics risk groups. AB - BACKGROUND AND OBJECTIVES: We studied the function of both Pgp and MRP1 to identify subgroups of patients who could benefit from Pgp reversion, and to clarify in different FAB subtypes and in cytogenetic risk groups their expression and function. DESIGN AND METHODS: We examined 132 adults with de novo acute myeloid leukemia (AML) for Pgp and MRP1 expression and function. We correlated our finding with the FAB subtypes and the cytogenetics, and clinical data of our patients. RESULTS: Among FAB subtypes and cytogenetic subgroups, patients with good risk cytogenetics have a low expression and activity of Pgp and MRP1 except patients with inv(16) who have a higher activity of MRP1 than t(8;21) and t(15;17) (p=0.05). All other AML patients, except M5, have a high expression and activity of Pgp. In contrast, M5 have a high expression, but a low activity of Pgp. In this subgroup, M5 with MLL gene rearrangement did not express an active Pgp. Others patients with M5 AML did not have a functional Pgp. Monosomy 7, 11q2.3 gene rearrangement and complex cytogenetic have a higher activity of MRP1 than other cytogenetic (p=0.03). INTERPRETATION AND CONCLUSIONS: Resistance mechanism in M5 was not mediated by Pgp. In contrast, MRP1 may play a role in patients who have a 11q2.3 gene rearrangement, or in M4E with inv(16). Thus trials that modulate Pgp are likely to achieve limited success in AML with low activity of Pgp, i.e., M5 and, AML with good risk cytogenetics. PMID- 14754605 TI - The suppressor of cytokine signaling-1 is constitutively expressed in chronic myeloid leukemia and correlates with poor cytogenetic response to interferon alpha. AB - BACKGROUND AND OBJECTIVES: Interferon-alpha (IFN-alpha) has proven useful for treating chronic myeloid leukemia (CML). However, only 7% of patients achieve a complete cytogenetic response. Although efforts to understand the molecular basis of this resistance to IFN-alpha have been made, the mechanism is still unknown. Because suppressor of cytokine signaling (SOCS) proteins are negative regulators of cytokine-induced signaling, it has been hypothesized that aberrant SOCS expression could confer resistance against cytokine therapy. DESIGN AND METHODS: In order to analyze the role of SOCS-1 in the acquisition of IFN-alpha resistance in this setting, we examined SOCS-1 mRNA expression using reverse transcription polymerase chain reaction (RT-PCR) in 75 newly diagnosed chronic phase-CML patients who received IFN-alpha therapy. RESULTS: SOCS-1 was constitutively expressed in 49 (65%) of 75 CML patients at diagnosis. Constitutive SOCS-1 expression was more frequently observed among Hasford high-risk patients (p = 0.05) and was also independently associated with a shorter median progression free survival time (p = 0.001) and poor cytogenetic response to IFN-alpha treatment (p 0.0001). INTERPRETATION AND CONCLUSIONS: Our data indicate that constitutive expression of SOCS-1 occurs at an early stage in CML pathogenesis and probably influences the clinical behavior of the disease. PMID- 14754606 TI - Quantitative reverse transcription polymerase chain reaction should not replace conventional cytogenetics for monitoring patients with chronic myeloid leukemia during early phase of imatinib therapy. AB - BACKGROUND AND OBJECTIVES: Imatinib is the new standard drug treatment for patients with chronic myelogenous leukemia (CML). Quantitative reverse transcription-polymerase chain reaction (qPCR) for detection of BCR-ABL transcripts is frequently used for monitoring patients in addition to or instead of conventional cytogenetics, although its place in the overall diagnostic framework is not yet clear. In this study, we compared qPCR and conventional cytogenetics for monitoring of patients during the early phases of imatinib therapy. DESIGN AND METHODS: One hundred and seventeen patients treated with imatinib for CML in chronic or accelerated phase were prospectively followed with qPCR and karyotyping. Comparisons were made between both methods and between qPCR results from bone marrow and peripheral blood. To determine the prognostic impact of qPCR and cytogenetics during the early phase of imatinib treatment on subsequent cytogenetic response and progression-free survival (PFS), a multivariate model was generated that included established prognostic baseline variables. RESULTS: We found a significant correlation between the proportion of Philadelphia (Ph) chromosome-positive metaphases and qPCR in the bone marrow and peripheral blood. Low qPCR values after 3 months of therapy were correlated with major cytogenetic response (MCyR) at 6 months and PFS at 2 years. However, in multivariate analysis, the cytogenetic response at 3 months emerged as the only independent parameter predictive of MCyR at 6 months and PFS at 2 years. INTERPRETATION AND CONCLUSIONS: Our data suggest that conventional karyotyping should remain the standard method for following patients on imatinib during the early phases of therapy. PMID- 14754607 TI - Incidence and characteristics of CD4(+)/HLA DRhi dendritic cell malignancies. AB - BACKGROUND AND OBJECTIVES: Recent reports suggest that CD4(+)/CD56(+)/lineage(-) hematopoietic neoplasias are aggressive types of malignancies involving lymphoplasmacytoid/DC2 dendritic cells (DC). Here, we report on the incidence of DC malignancies and their clinical, biological, phenotypic and cytogenetic characteristics. DESIGN AND METHODS: From a large series of 392 patients with acute myeloblastic leukemia (AML) and 739 with non-Hodgkin's lymphoma (NHL), five cases (three presenting as acute leukemia and two as NHL) showed clinical, morphologic and phenotypic features compatible with a DC malignancy. RESULTS: The overall incidence of DC malignancies among all AML and NHL cases was 0.76% and 0.27%, respectively. At presentation, these patients displayed cutaneous nodules, splenomegaly and lymph node involvement with variable levels of peripheral blood (PB) and/or bone marrow (BM) infiltration in association with anemia and thrombocytopenia. Cytologic studies showed immature appearing blast cells with negative cytochemistry reactions for both myeloperoxidase and esterases. A highly suggestive DC phenotype based on co-expression of CD123(hi)/HLADR(+)/lin( )/CD56(+)/CD45(dim) associated with a germline configuration of both the IgH and TCRgamma genes was found in all except one patient who was CD56(-). Expression of other markers compatible with a DC origin was found in all cases. INTERPRETATION AND CONCLUSIONS: We show that DC-derived malignancies can present as either cutaneous lymphoma or acute leukemia, although their incidence is extremely low (<< >1%). While most of these DC neoplasias probably correspond to the malignant counterpart of DC2/lymphoplasmacytoid DC, neoplasias of myeloid DC might also exist, chemotherapy followed by consolidation with ASCT is apparently the most effective strategy for achieving a durable remission in these individuals. PMID- 14754608 TI - Prevalence of hepatitis C virus infection in lymphoproliferative diseases other than B-cell non-Hodgkin's lymphoma, and in myeloproliferative diseases: an Italian Multi-Center case-control study. AB - BACKGROUND AND OBJECTIVES: Infection with hepatitis C virus (HCV) is associated with type II mixed cryoglobulinemia (MC), a lymphoproliferative disorder which, in some patients, evolves into overt B-cell non-Hodgkin's lymphoma (B-NHL). Recently, also the association between HCV infection and B-NHL, which had long been controversial, was confirmed in a large case-control study. Little knowledge is, however, available on possible associations between HCV infection and other lymphoid or myeloid malignancies. The present study was set up in order to investigate this aspect. DESIGN AND METHODS: The study was conducted in hematology departments of ten hospitals in different Italian cities. The cases consisted of consecutive patients with a new diagnosis of T-NHL, Hodgkin's disease (HD), chronic lymphocytic leukemia (CLL), acute lymphoblastic leukemia (ALL), multiple myeloma (MM), acute myeloid leukemia (AML), and chronic myeloid leukemia (CML). The controls were patients in other departments of the same hospitals. HCV infection was investigated by testing for HCV antibodies and HCV RNA in serum samples. RESULTS: The prevalence of HCV infection was not higher in patients with HD (3.2%, 5 out of 157 cases) or MM (4.7%, 5 out of 107) than in controls. On the other hand, it was consistently higher in T-NHL (13.8%, 4 out of 30), CLL (9.0%, 9 out of 100), ALL (7.6%, 5 out of 54), AML (7.9%, 11 out of 140), and CML (12.2%, 6 out of 49) patients. These patient groups were not, however, large enough to render statistically significant results. INTERPRETATION AND CONCLUSIONS: Our data suggest that HCV infection may be associated not only with B-NHL but also with some other lymphoid and myeloid malignancies. PMID- 14754609 TI - A sensitive ristocetin co-factor activity assay with recombinant glycoprotein Ibalpha for the diagnosis of patients with low von Willebrand factor levels. AB - BACKGROUND AND OBJECTIVES: The assay of ristocetin co-factor activity of von Willebrand factor (VWF:RCo) is used in the screening of patients with suspected von Willebrand's disease (VWD), the most frequent inherited bleeding disorder. A correct diagnosis of VWD relies on platelet agglutination tests that have a low accuracy within and between assays. A more accurate VWF:RCo assay would improve VWD diagnosis and classification. DESIGN AND METHODS: We describe here an ELISA method in which a recombinant fragment of the alpha-subunit of platelet glycoprotein Ib-IX-V complex (rGPIbalpha) is bound to an anti-GPIbalpha monoclonal antibody immobilized onto microtiter plate wells and which captures plasma VWF in the presence of ristocetin. The results obtained with this ELISA assay were compared blindly with values calculated from the agglutination test in normal subjects (n=60) and in type 1 (n=8), type 2A (n=16), type 2B (n=13), type 2M (n=17) or type 2M Vicenza (n=8) VWD patients which were characterized by low VWF levels. RESULTS: The two assays gave similar results in both normal subjects and VWD patients (r=0.93), but the ELISA test showed higher sensitivity (0.1 versus 6.25 U/dL). The repeatability and reproducibility gave coefficients of variation of 9% and 10%, respectively, for the ELISA, as compared to 14% and 15% for the agglutination test. INTERPRETATION AND CONCLUSIONS: This ELISA assay can be useful in the identification and classification of VWD patients in that it may provide a more accurate distinction between type 2 with abnormal VWF function and type 1 with a low plasma VWF concentration. PMID- 14754610 TI - Elongation factor 1 (EF1alpha) promoter in a lentiviral backbone improves expression of the CD20 suicide gene in primary T lymphocytes allowing efficient rituximab-mediated lysis. AB - BACKGROUND AND OBJECTIVES: CD20 has been proposed as a novel suicide gene system for the treatment of graft-versus-host disease (GVHD), a fatal complication of allogeneic bone marrow transplantation: indeed expression of the human non immunogenic exogenous CD20 protein allows positive immunoselection of transduced cells as well as their killing in vitro with rituximab. Lentiviral vectors are promising tools in the field of gene therapy. We therefore searched for a lentivector giving good efficiency of transduction of human T lymphocytes activated by the sole addition of interleukin (IL)-2 and high expression levels of the CD20 transgene. DESIGN AND METHODS: The T cell line CEM and peripheral T lymphocytes activated by phytohemagglutinin (PHA) and/or IL-2 were transduced with two different vectors carrying the CD20 transgene driven by either the phosphoglycerate kinase (PGK) or elongation factor 1alpha (EF1alpha) promoter, and using different multiplicities of infection (MOIs). RESULTS: Both the PGK- and EF1alpha-CD20 vectors allowed efficient transduction of the CEM cell line and PHA-activated T cells, reaching 99 and 90% in the different targets, respectively. However EF1alpha-CD20 led to much higher expression levels of the transgene (mean fluorescence intensity 588-618 compared to 53 for PGK-CD20). Furthermore lymphocytes activated with IL-2 alone could be efficiently transduced with EF1alpha-CD20, reaching 10-25% positivity for CD20 (mean fluorescence intensity 409-424), allowing adequate immunoselection and strong complement mediated lysis. INTERPRETATION AND CONCLUSIONS: EF1alpha-CD20 may represent a good candidate vector for gene therapy with the CD20 suicide system in the setting of allogeneic bone marrow transplants. PMID- 14754611 TI - Acquired hemophilia: a critical bleeding syndrome. PMID- 14754612 TI - Detection of intracellular phosphorylated ERK1/2 in natural killer cells by flow cytometry. PMID- 14754613 TI - Patterns of AML1-ETO transcript expression in patients with acute myeloid leukemia and t(8;21) in complete hematologic remission. PMID- 14754614 TI - Second hit mutations in the RTK/RAS signaling pathway in acute myeloid leukemia with inv(16). PMID- 14754615 TI - Comparison of BAVC to BuCy regimens in autologous stem cell transplantation for adult patients with acute myeloid leukemia. PMID- 14754616 TI - ATM mutations in B-cell chronic lymphocytic leukemia. PMID- 14754617 TI - Dendritic cell deficiency in early B cell chronic lymphocytic leukemia. PMID- 14754618 TI - Splenic marginal zone lymphomas with peripheral CD5 expression. PMID- 14754619 TI - Positron emission tomography with [18F] 2-fluoro-D-2- deoxyglucose in primary cutaneous T-cell lymphomas. PMID- 14754620 TI - Identification of six novel mutations in type I antithrombin deficient Italian families. PMID- 14754621 TI - High factor VIII plasma levels as a risk factor for venous thrombosis: no evidence of inheritance from a family cohort study. PMID- 14754622 TI - Immunological reconstitution after autologous peripheral blood stem cell transplantation in patients with chronic lymphocytic leukemia. Comparison with an historical non-Hodgkin's lymphoma group. PMID- 14754623 TI - Hematopoietic abnormalities persist for more than six years after autologous peripheral blood stem cell transplantation in patients with non-Hodgkin's lymphoma. PMID- 14754624 TI - Investigation of health perspectives of those with physical disabilities: the role of spirituality as a determinant of health. AB - PURPOSE: To identify key determinants of health and the process of health attainment for people with musculoskeletal disabilities. METHOD: Focus groups of people with musculoskeletal disorders, including 30 members and their five trained facilitators, provided data. Discussed were 'What is health for you?' and 'What has helped, or would help you achieve this health?' Delphi-structured analysis identified health themes and a health process model was developed with the facilitators comprising the expert panel. RESULTS: Health was perceived as centred on relationships that required a spiritual awareness for a strong and resilient identity. The Self Attributes Model developed portrays the processes perceived to be required for health. CONCLUSIONS: Although physical, social and psychological interventions are essential aspects of health intervention, by themselves they are not sufficient. Also required for health is a strong resilient self resulting from interaction and connection with other people and the natural world. Moreover, development of such an identity requires a spiritual world-view comprising an acknowledgement of the essence of self and focus upon the nature of the connection of this essence with all other aspects of life. Further research is required to advance understanding of the process by which this occurs for people with chronic disorders. PMID- 14754625 TI - Vocational rehabilitation acceptance in the USA: controlling for education, type of major disability, severity of disability and socioeconomic status. AB - PURPOSE: The aim of the study was to investigate whether there were differences in acceptance rates for VR services among African Americans, White Americans, Native American or Alaskan Natives, and Asian or Pacific Islanders with disabilities in the USA? METHOD: The study was based on a population 599 444 customers who sought VR or Bureau of Visual Service Agency services in the USA from 1 October, 1997, through 30 September, 1998. The subsample of customers with no missing values on the variables under investigation included African Americans (n = 13 287), White Americans (n = 38 048), Native American or Alaskan Natives (n = 599), and Asian or Pacific Islanders (n = 596). The chi-square test of homogeneity of proportions was the test statistic. The final random subsample included African Americans (n = 300), White Americans (n = 300) Native American or Alaskan Natives (n = 300), and Asian or Pacific Islanders (n = 300) was drawn from the population of VR customers in the USA. RESULTS: The study supports the hypothesis that African Americans were more likely to be found ineligible for VR services, while Asian or Pacific Islanders were more likely to be accepted for VR services. CONCLUSION: While discovering that African Americans are more likely to be rejected for VR services was not surprising, discovering that Asians or Pacific Islanders are more likely to be accepted for VR services than African Americans was unexpected, given that past VR acceptance research adduced that White Americans, not Asian or Pacific Islanders, are more likely to be accepted for VR services when compared to African Americans with disabilities. While a preponderance of VR research indicates that White Americans are more likely to be accepted for VR services than African Americans, it was also unexpected that White Americans were not statistically significant when education, type of major disability, disability severity, and SES were controlled. PMID- 14754626 TI - The effect of cane length on the haptic perception of height. AB - PURPOSE: The purpose of this investigation was to determine whether the ability to haptically perceive object height is a function of the length of cane being wielded by blindfolded individuals. METHOD: Fifteen sighted females between the ages of 18 and 25 years volunteered to participate in the study. Five different heights of wooden blocks were haptically explored with four different cane lengths (0.92, 1.22, 1.37 and 1.67 m), cane length order being randomized across 2 days of testing. Subjects were asked to report whether the explored block's height was taller, shorter, or equal to that of a previously presented standard block. The percentages of correct judgments for each cane and block height combination were subjected to statistical analysis. RESULTS: The analysis revealed significant main effects for cane length, F (3, 42) = 9.47, p < 0.0001, and block height, F (4, 56) = 17.69, p < 0.0001. The interaction of cane length and block height was not significant, F < 1. The accuracy of haptic perception improved with decreased cane length and increased difference in height from the standard block. CONCLUSIONS: Contrary to previous research on haptic perception in which probe length was found not to affect judgment accuracy, the present study found a marked influence of cane length on perceptual accuracy. This finding indicates that length is an important property that should be taken into account when prescribing canes for the visually impaired. PMID- 14754627 TI - Reliability and stability of the Roland Morris Disability Questionnaire: intra class correlation and limits of agreement. AB - PURPOSE: To analyse test-retest reliability and stability of the Dutch language version of the Roland Morris Disability Questionnaire (RMDQ) in a sample of patients (n = 30) suffering from Chronic Low Back Pain (CLBP). METHOD: Patients filled out the Dutch language version of the RMDQ questionnaire twice, before starting the rehabilitation programme, with a 2-week interval. Intra Class Correlations (ICC), (one way random) was used as a measure for reliability and the limits of agreement were calculated for quantifying the stability of the RMDQ. An ICC of 0.75 or more was considered as an acceptable reliability. No criteria for limits of agreement were available. However, smaller limits of agreement indicate more stability because it indicates that the natural variation is small. RESULTS: The Dutch RMDQ showed good reliability, with an ICC of 0.91. Calculating limits of agreement to quantify the stability, a large amount of natural variation ( +/- 5.4) was found relative to the total scoring range of 0 to 24. CONCLUSION: The Dutch RMDQ proves to be a reliable instrument to measure functional status in CLBP patients. However, the natural variation should be taken into account when using it clinically. PMID- 14754628 TI - The importance of cultural factors in the planning of rehabilitation services in a remote area of Papua New Guinea. AB - PURPOSE: The aim of this study in the Middle Ramu, Papua New Guinea, was to gain a better understanding of how cultural factors work to influence the lives of persons with disability in a remote area. The study also explores how this information can be used for the planning of rehabilitation services. METHOD: Two phase screening identified persons with disability in the study area and questionnaires were completed for all those identified. Information documented included the nature of the disability, a biomedical cause (where appropriate), the perceived cause of the disability, as well as some indication as to where help had been sought for the disability. In depth interviews were later done with disabled individuals and their families, to determine how they explained their disability. RESULTS: Thirty-two per cent of persons with disability and their families attributed disability to sorcery or other supernatural causes, a greater proportion than for any other category of perceived aetiology. There was widespread acceptance of Western medicine, although help was more likely to be sought from sources in the community for disabilities believed to have a supernatural origin. CONCLUSIONS: This study demonstrates that an understanding of cultural factors is fundamental to implementing rehabilitation services that are culturally appropriate and address the social dimension of disability. PMID- 14754629 TI - Functional independence of residents in urban and rural long-term care facilities in Taiwan. AB - PURPOSE: To compare the score of functional independence measure (FIM) between urban and rural residents living in long-term care facilities (LTCF) in Taiwan. METHODS: A total of 437 subjects in 112 licensed LTCF in Taiwan were randomly selected by stratification strategy. Physical therapists interviewed the subjects in nursing homes (NH) and intermediate care facilities (ICF) to obtain the basic data, and the FIM score. RESULTS: (1) There was no significant difference in basic demographic data between urban and rural LTC subjects. (2) Most of the subjects in urban and rural LTCF were males, less than 80 years old, single/widowed, having multiple diseases, using more than one assistive devices, and having social welfare financial support. (3) Motor abilities (eating, grooming, and transfer) and cognition (comprehension, social interaction and problem solving) in rural LTCF subjects were significantly (p < 0.05) higher than those in urban areas as revealed by the FIM assessment. (4) The median of FIM total score of rural LTCF subjects was 90.5, which was significantly (p < 0.05) higher than that of urban LTCF subjects (median = 76). CONCLUSIONS: Some of the functional performance of subjects in rural long-term care institutions is better than those in urban areas. Our results may provide guidelines for the manpower and equipment supply estimation. PMID- 14754630 TI - The World Health Organisation's terminology and classification: application to severe disability. AB - PURPOSE: The purpose of this article is to describe the international classification system proposed by the World Health Organisation for describing individuals with disability. Initially the 'International classification of impairment, disability and handicap' (ICIDH) was used. This has been replaced by the 'International classification of functioning, disability and health' (ICF). Both of these systems will be described and followed by a discussion of the advantages and disadvantages of using the WHOs classification framework. An application to the field of severe disability will be made throughout. METHOD: Providing a theoretical framework for classification of disability in accordance with the system proposed by the WHO. RESULTS: The ICF is a useful tool that contributes to uniformity of international terminology and standardization in the disability field. It is not a minority model, and focuses on strengths and skills. CONCLUSIONS: An international classification system such as the ICF offers a conceptual framework for information that is relevant to the long-term consequences of disability. Although any type of classification system has certain limitations, the advantages present within the ICF outweigh the limitations. PMID- 14754633 TI - Chemical interactions between additives in foodstuffs: a review. AB - This paper critically reviews the key literature on food additive-additive chemical interactions published over the last 30 years together with appropriate relevant information on food additive-food component interactions. Five main classes of food additive are included, reflecting the research effort to date: the sulfur (IV) species of preservatives, synthetic food colouring materials, nitrate and nitrite, ascorbic acid, and sorbic acid. Within each class, aspects of the chemistry (reactivity), functionality, stability, use and reactions with other specific food additives are reviewed. Where appropriate, the importance of interactions of food additives with other components of food (i.e. nutrients and non-nutrients) has been assessed and certain aspects of toxicology included. The practical outcome of this review is presented as a set of recommendations for future research in this area. The use of the data in this review is proposed as a training set to develop the framework into a diagnostic tool. This might be used ultimately for the development of a multilevel framework, operating systematically, to understand the important parameters that dictate the outcome of additive interactions. PMID- 14754634 TI - Novel method for the determination of added annatto colour in extruded corn snack products. AB - There is considerable interest in determining the added levels of the natural dye annatto in foods like snack products, particularly because they are mostly consumed by young people. The objective was to use response surface methodology to develop a new method to analyse annatto in extruded snacks. A pretreatment of the samples was necessary, digesting the ground sample with alpha-amylase at room temperature. The pigment was extracted by shaking with ethyl acetate at room temperature, eight extractions being necessary for completion extract the pigment. Lipids were removed by alkaline saponification. Under these conditions, 100% of the bixin was converted into norbixin, which was then quantified by high performance liquid chromatography. The method had a mean recovery of 97% and a coefficient of variation for duplicate analysis of 1%. Using this method, of the 13 commercial samples analysed, a parmesan cheese-flavoured snack product showed the highest level of dye expressed as norbixin (15.5 mg kg(-1)), whilst other brands of onion-flavoured snack products had the lowest levels (0.7 and 0.4 mg kg(-1), respectively). PMID- 14754635 TI - Xenoestrogens and the induction of proliferative effects in breast cancer cells via direct activation of oestrogen receptor alpha. AB - Environmental contamination with a variety of industrial products has been associated with developmental and reproductive abnormalities in wildlife species. Increasing evidence has suggested that bisphenol A (BPA) and 4-nonylphenol (NPH), two major endocrine-disrupting chemicals, might be responsible for adverse effects on humans as a consequence of ubiquitous use together with potential oestrogen-like activity. To provide insight into the oestrogen-like nature of BPA and NPH, their ability to activate a reporter gene construct via an oestrogen response element in the hormone-dependent breast cancer cell lines MCF7 and T47D was ascertained. Both compounds transactivated the endogenous oestrogen receptor (ER) alpha in a direct fashion since the anti-oestrogen 4-hydroxytamoxifen abolished the response. In addition, using steroid-receptor-negative HeLa cells engineered to express ERalpha and ERbeta and the hormone-binding domains of both ERalpha and ERbeta, BPA and NPH confirmed the direct transcriptional activity. Interestingly these properties were supported in MCF7 cells by the ability to autoregulate ERalpha expression as well as to induce its nuclear compartmentalization. We therefore evaluated by reverse transcriptase polymerase chain reaction the expression of oestrogen-controlled genes such as cathepsin D and TFF1 (formerly pS2), which were increased by both chemicals tested. The agonistic effects exhibited in all assays performed prompted the evaluation of a more complex biological response such as the proliferation of MCF7 and T47D cells. The same concentration of xenoestrogens eliciting substantial transcriptional activity significantly stimulated the proliferation of both breast cancer cell lines, although with a reduced effectiveness with respect to the natural hormone 17beta-oestradiol. The results indicate that the biological action of environmental oestrogen such as BPA and NPH should be taken into account for the potential impact on human disease-like hormone-dependent breast cancer. However, further studies are needed to clarify their bioavailability and metabolism as well as whether compound mixtures could produce noticeable effects by synergistic activity. PMID- 14754636 TI - Evaluation and establishing the performance of different screening tests for tetracycline residues in animal tissues. AB - Four methods intended for screening muscle tissue for residues belonging to the tetracycline group were compared using artificially contaminated as well as incurred samples. Two agar diffusion methods were studied: one with Bacillus subtilis as a test strain, the second with Bacillus cereus. Two variants of each method were compared: thin plates for analysis of intact or minced meat, and thick plates for analysis of meat fluid. The thin plate variants could not be evaluated with artificially contaminated samples because it was impossible to prepare homogeneously spiked, undiluted meat. The thick plates were suited for doxycycline and chlortetracycline, but they did not detect oxytetracycline or tetracycline in spiked meat fluid. The results of these tests done on incurred meat were very good for doxycycline and satisfying or just failing for oxytetracycline, while the best detection capability was obtained when intact frozen meat was examined on thin plates seeded with B. cereus. Two commercially available screening tests were also evaluated. The Premi(R) test, an inhibitor test with Bacillus stearothermophilus as a test strain and an indicator for growth, was not suited for detection of tetracyclines up to the maximum residue limit. Tetrasensor(R), a receptor test specific for tetracyclines, proved a quick and simple test able to detect meat samples artificially contaminated with tetracycline, oxytetracycline, doxycycline or chlortetracycline, as well as meat incurred with oxytetracycline or doxycycline. PMID- 14754637 TI - Determination of lead and cadmium in wines by graphite furnace atomic absorption spectrometry. AB - The lead (Pb) and cadmium (Cd) content of various wines on the Korean market were determined by graphite furnace atomic absorption spectrometry using Zeeman background correction and peak area mode. All wine samples were microwave digested in concentrated HNO(3). Ammonium dihydrogen phosphate and magnesium nitrate were used as matrix modifiers for both Pb and Cd analyses. The mean Pb content of the wines was about 29 microg l(-1) ranging from 5 to 87 microg l(-1). Also, the means of Cd were about 0.5 microg l(-1) ranging from < 0.1 to 3.0 microg l(-1). The mean recoveries of Pb and Cd were 92.8 and 101.3% and their analytical detection limits were 1.0 and 0.1 microg l(-1), respectively. Sixty brands of wine were classified into red and white, but no statistically significant difference in Pb and Cd content was observed. PMID- 14754638 TI - Binding of aflatoxin B1 to cell wall components of Lactobacillus rhamnosus strain GG. AB - The surface of Lactobacillus rhamnosus strain GG (LGG) has previously been shown to bind aflatoxin B(1) (AFB(1)) effectively, it being a food-borne carcinogen produced by certain species of Aspergillus fungi. To establish which components of the cell envelope are involved in the AFB(1) binding process, exopolysaccharides and a cell wall isolate containing peptidoglycan were extracted from LGG and its AFB(1) binding properties were tested. LGG was also subjected to various enzymatic and chemical treatments and their effects on the binding of AFB(1) by LGG were examined. No evidence was found for exopolysaccharides, cell wall proteins, Ca(2+) or Mg(2+) being involved in AFB(1) binding. The AFB(1) binding activity of the cell wall isolate indicates that AFB(1) binds to the cell wall peptidoglycan of LGG or compounds tightly associated with the peptidoglycan. PMID- 14754639 TI - Limited survey for the presence of aflatoxins in foods from local markets and supermarkets in Valencia, Spain. AB - Aflatoxin B1 (AFB1), B2 (AFB2), G1 (AFG1) and G2 (AFG2) were extracted by matrix solid-phase dispersion with C18 silica and acetonitrile as the eluting solvent, analysed by liquid chromatography with fluorescence detection and confirmed by liquid chromatography with mass spectrometry using an electrospray interface in 58 samples grouped as cereals, dried fruits, herbs and spices, pulses, snacks, and nuts and nut products collected from local markets and supermarkets in Valencia, Spain. All samples analysed by the proposed method were previously studied with an enzyme-linked immunosorbent assay as a screening protocol for the fast detection of mycotoxins. The samples containing residues (3/58) were hazelnut (0.42 and 0.52 microg kg(-1) for AFB1 and AFG1, respectively), nut cocktail (0.29 and 0.47 microg kg(-1) for AFB1 and AFG1, respectively) and pinhol (0.30 microg kg(-1) for AFG1). Such values were below the legislated maximum residue levels for the European Union. PMID- 14754640 TI - Survey of ochratoxin A and deoxynivalenol in stored grains from the 1999 harvest in the UK. AB - Three hundred and twenty samples from the 1999 UK harvest comprising wheat (201 samples), barley (106) and oats (13) were analysed for ochratoxin A and deoxynivalenol. A small number of organic samples was also obtained. Samples were collected from farms, central stores, mills, maltings and ports from across the UK from February to April 2000. Ochratoxin A and deoxynivalenol analysis was by affinity column clean up and high-performance liquid chromatography with fluorescence and ultraviolet light detection, respectively, with limits of detection of 0.2 and 20 microg kg(-1). The survey found ochratoxin A at below 5 microg kg(-1) in 97% of the samples indicating satisfactory storage conditions. The remaining 3% of the samples contained ochratoxin A at levels between 5.2 and 231 microg kg(-1), but none of these samples was intended for human consumption. Deoxynivalenol was detected in 88% of all samples, with 83% below 100 microg kg( 1); the maximum level was 600 microg kg(-1). Twenty samples containing deoxynivalenol at or above 150 microg kg(-1) by high-performance liquid chromatography were all confirmed by gas chromatography/mass spectrometry. Nivalenol was also detected by gas chromatography/mass spectrometry at levels of 50 microg kg(-1) or higher in 18 of 20 samples where deoxynivalenol was confirmed. PMID- 14754641 TI - Levels of polychlorinated dibenzo-p-dioxins and dibenzofurans in cattle raised at agricultural research facilities across the USA and the influence of pentachlorophenol-treated wood. AB - Adipose tissue samples from 158 cattle raised locally at experiment stations across the USA were analysed for polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/F). While 80% of the samples had PCDD/F concentrations that fell within the range of a previous US survey of beef animals (not detected -4.1 ppt toxic equivalency), several animals had exceptionally high concentrations (8 54 ppt toxic equivalency). The investigations of three facilities where highly contaminated animals were raised found pentachlorophenol-treated wood at each site. The congener pattern in the animals' tissues and the lack of elevated PCDD/F levels in other environmental samples, i.e. hay and soil, indicated that the treated wood was the source of contamination. A congener pattern similar to that of pentachlorophenol-exposed animals was seen for the means and medians of the entire data, i.e. OCDD, HpCDD and 1,2,3,6,7,8-HxCDD dominated, the PCDD concentrations equalled or exceeded the furan concentrations, and the concentration of 1,2,3,6,7,8-HxCDD was six times that of the other HxCDD isomers. This suggested that pentachlorophenol-treated wood contributed measurably to many of the animals in this survey. The largest contributors to the median toxic equivalencies were 1,2,3,7,8-PeCDD (40%) and 1,2,3,6,7,8-HxCDD (16%). No clear geographical trends emerged from the data. PMID- 14754642 TI - Friends old and new. PMID- 14754643 TI - Single-center retrospective study of long-term use of low-dose acitretin (Soriatane) for psoriasis. AB - BACKGROUND: Indicated as monotherapy for severe psoriasis in adults, acitretin (Soriatane) can be used in combination therapy or as a 'disease-stabilizing, maintenance' agent. While its efficacy for psoriasis and other disorders of keratinization is well established, its safety is often a concern for many dermatologists. The possible side effects associated with short-term treatment of oral retinoids include mucocutaneous effects, elevation in serum lipid chemistries and liver enzymes, and teratogenicity. However, the only possible long-term, cumulative side effect is skeletal and ligamentous calcification such as hyperostosis. There is a specific syndrome of hyperostosis associated with oral retinoids called DISH (diffuse idiopathic skeletal hyperostosis). OBJECTIVE: To examine the incidence of DISH syndrome in long-term acitretin use for psoriasis and to quantify the incidence of other side effects. METHODS: All patients seen at the UCSF Psoriasis and Skin Treatment Center who had been on acitretin for more than 1 year were identified. RESULTS: A chart review of these patients revealed no X-ray-confirmed cases of DISH syndrome, minimal changes in coronary heart disease risk indicators, and extremely rare significant elevation of liver enzymes. CONCLUSION: Acitretin does not appear to cause significant long term side effects at low doses; the implications for thousands of patients are that use of this medication can be continued for long periods of time with routine monitoring. PMID- 14754644 TI - Double-blind, randomised, multicentre, parallel group study comparing a 1% coal tar preparation (Exorex) with a 5% coal tar preparation (Alphosyl) in chronic plaque psoriasis. AB - BACKGROUND: Exorex lotion is a novel formulation of prepared coal tar indicated for the treatment of psoriasis. OBJECTIVES: To compare the efficacy and tolerability of 1% prepared coal tar lotion versus 5% coal tar extract in patients with mild to moderate plaque psoriasis. PATIENTS AND METHODS: This was a double-blind, randomised controlled study. Patients initially entered a 7-day washout period, during which they applied a yellow soft paraffin plus emulsifying wax ointment used as an emollient three times a day to their plaques. They were then randomised to receive treatment with 1% coal tar (Exorex) lotion or 5% conventional coal tar lotion (Alphosyl), three times a day for 12 weeks. Both treatment groups continued to apply the emollient throughout the duration of the study. Two target plaques were selected at entry for assessment. The clinical measures used were: 1) Total Sign Score (TSS), the sum of 5-point rating scores for erythema, induration and scaling averaged for the two target plaques (range 0 12), 2) the Psoriasis Area and Severity Index (PASI), and 3) patient and investigator 7-point global assessments of improvement at 12 weeks. Patients were assessed at 0, 4, 8 and 12 weeks during the treatment period or at the point of withdrawal. Spontaneously reported and observed adverse events were noted. RESULTS: Three hundred and twenty four of 338 randomised patients were evaluable (ITT analysis): 158 patients received 1% coal tar lotion and 166 patients received conventional coal tar. Both groups showed decreases from baseline to end of treatment in mean TSS (decrease of 2.4 points from 5.6 to 3.2 with 1% coal tar lotion and 1.8 points from 5.5 to 3.7 with conventional coal tar), and mean PASI (decrease of 2.4 points with 1% coal tar lotion and 1.5 points with conventional coal tar). Two hundred and twenty eight patients completed the full course of treatment. There was a statistically significant treatment difference in the percentage change in mean TSS at week 12, in favour of 1% coal tar lotion ( 10.6%, 95% CI -20.6% to -0.5%, p=0.04). There was also a difference between treatments in the change in mean PASI in favour of 1% coal tar that was of borderline statistical significance (-11.7%, 95% CI -23.8% to 0.4%, p=0.06). Investigator global assessments also favoured 1% coal tar lotion (38% vs. 27% of patients showed clearance or marked improvement). The 1% coal tar lotion had a similar safety profile to 5% conventional coal tar lotion with the majority of treatment-related events being mild to moderate in severity. CONCLUSIONS: 1% coal tar lotion is more effective than a conventional coal tar lotion in mild to moderate psoriasis and may be preferred for first-line topical treatment. PMID- 14754645 TI - Fumaric acid esters in the treatment of psoriasis: an Italian experience. AB - BACKGROUND: Psoriasis is a common, chronic, cell-mediated, inflammatory skin disease. Treatment limitations and a developing understanding of its pathogenesis on a molecular level have encouraged much interest in the field of immunomodulatory therapy. OBJECTIVE: To evaluate the efficacy and safety of fumaric acid esters, in particular dimethylfumarate (DMF), in the treatment of moderate to severe plaque psoriasis intolerant and/or resistant to other conventional systemic therapies. METHODS: A total of 40 patients were enrolled in this study. DMF was orally administered at the daily dose of 30 mg up to 360 mg for a minimum of 6 months treatment. Patients were followed-up with psoriasis area and severity index (PASI) score assessment, and clinical and photographic documentation. RESULTS: A total of 33 (82.5%) patients achieved complete clinical remission with DMF treatment: eight after 3 months and 25 after 6 months. Adverse events, such as intolerable abdominal cramps and incoercible diarrhoea, occurred in four patients who, for this reason, interrupted therapy. CONCLUSION: The findings suggest that DMF is a safe, effective and well-tolerated long-term oral treatment worthy of consideration for selective patients. PMID- 14754646 TI - A quantitative definition of severe psoriasis for use in clinical trials. AB - Clinical trials of new agents for severe psoriasis require clear, well characterized and objective measurement criteria to define the study population. The purpose of this paper is to offer parameters defining 'severe' psoriasis for use in clinical trials. If patients have at least 10% body surface area (BSA) involved and a psoriasis area and severity index (PASI) score > or =12, they have severe disease. While this definition assures inclusion of only patients with severe disease, a disadvantage is that some patients with truly severe disease (as defined by the impact of their disease on their quality of life) will be excluded. It is necessary to accept this trade off in order to have objective criteria, consistent across centers involved in clinical trials, for identifying patients affected by severe psoriasis. PMID- 14754647 TI - A multicenter trial of the efficacy and safety of 0.03% tacrolimus ointment for atopic dermatitis in Korea. AB - BACKGROUND: Atopic dermatitis is a chronically relapsing, common inflammatory skin disease, which significantly affects quality of life negatively in many respects. Topical steroids are the mainstay of atopic dermatitis treatment but they carry the risk of local side effects. A topical formulation of tacrolimus, a macrolide calcineurin inhibitor, has recently been developed. OBJECTIVE: To evaluate the efficacy and safety of 0.03% tacrolimus ointment for the treatment of moderate to severe atopic dermatitis in Korea. METHODS: An open, non comparative, multi-center study with 4 weeks' follow-up was performed. A total of 180 patients (aged 2-57 years old) were enrolled. Tacrolimus ointment (0.03%) was applied to all involved areas twice daily. Efficacy was evaluated by an investigator's global assessment, the eczema area and severity index score, and by the patient's assessment of pruritus and clinical response at baseline, and after weeks 1, 2 and 4. Dermatology life quality index (DLQI), children's DLQI (CDLQI) and toddler's DLQI were assessed at baseline and at week 4. The safety assessment included monitoring all adverse events and clinical laboratory values. RESULTS: All efficacy parameters were improved. The mean EASI (eczema area and severity index) score was 19.7 at baseline and reduced to 8.0 at the end of the study. Moderate improvement was observed by the investigator's global assessment after 4 weeks' treatment. A marked decrease of pruritus was observed, and mild or moderate improvement was observed by patients' global assessments after the treatment period. Significant benefits in terms of quality of life in adults and children with atopic dermatitis were obtained. The most common adverse events associated with tacrolimus treatment were transient skin burning sensation (45.3%) and pruritus (41.6%) at the site of application. CONCLUSION: 0.03% tacrolimus ointment should be considered effective and safe in both Korean children and adults with moderate to severe atopic dermatitis. PMID- 14754648 TI - Topical tacrolimus 0.1% ointment for refractory skin disease in dermatomyositis: a pilot study. AB - BACKGROUND: Dermatomyositis is an inflammatory disease primarily involving the striated muscles and skin. Muscle disease usually responds to aggressive therapy with systemic corticosteroids. However, cutaneous lesions can be very resistant to systemic and topical therapies, even in combination. More treatment options are needed. Tacrolimus is an immunomodulator now available in a topical ointment. OBJECTIVE: To study the treatment of patients with refractory cutaneous lesions of dermatomyositis using topical tacrolimus 0.1% ointment. METHODS: Six patients with recalcitrant cutaneous lesions of dermatomyositis were included in this brief observational study: five adults and one pediatric patient. Five patients had classic dermatomyositis and one had dermatomyositis sine myositis. RESULTS: All had some degree of improvement of their dermatologic disease following 6-8 weeks of treatment with topical tacrolimus 0.1% ointment. Two had dramatic responses (>90% improvement), one had moderate (40-90%) improvement and three had minimal (20-40%) improvement. CONCLUSIONS: The dermatologic manifestations of dermatomyositis can be very difficult to treat. Multiple systemic and topical therapies have been studied. Combinations of treatments are often used, sometimes still not successfully. The results of this brief observational study using topical tacrolimus ointment for the treatment of refractory cutaneous lesions of dermatomyositis are encouraging. Topical tacrolimus was a useful adjunct in the treatment of the dermatologic component of dermatomyositis in adults and children in this pilot study. PMID- 14754649 TI - Pityriasis versicolor: quality of studies. AB - OBJECTIVE: To evaluate the quality of the published literature on the treatment of pityriasis versicolor and to compare this evaluation with the number of times the work has been cited. METHODS: A search was performed using MEDLINE (1966 2002) to find publications evaluating the treatment of pityriasis versicolor. Two reviewers evaluated each clinical trial according to a list of pre-determined criteria including randomization and blinding, prior sample size calculation, treatment regimen clearly explained, and well-defined efficacy parameters. A maximum score of 20 could be attained by each publication. A citation count was performed using the ISI Web of Science Database (http://www.isinet.com/isi/products/citation/wos/). RESULTS: A total of 94 studies were included in this analysis of quality. Studies with a score of 8 or more were considered to be high quality, and 61 studies were rated as high quality studies (65%). There was no significant correlation between paper quality and citation count. CONCLUSION: It is important for investigators and clinicians to be aware of the design of a high-quality protocol, since such high-quality studies are more likely to reflect accurate efficacy rates. Also, it is important for the information on the design and conduct of the trial to be conveyed to the reader so that a meaningful comparison between the studies can be made. PMID- 14754650 TI - Intravenous immunoglobulin (IVIg) for therapy-resistant cutaneous lupus erythematosus (LE). AB - BACKGROUND: A group of patients exists with predominantly cutaneous lupus erythematosus (LE) who do not respond to standard oral therapies. There has been interest in the role of intravenous immunoglobulin (IVIg) in a number of connective tissue diseases, and its role in some circumstances has been proven. In the case of LE, there are suggestions that the use of IVIg for cutaneous and more systemic disease may be of value. OBJECTIVE: To investigate the use of low dose IVIg for therapy-resistant cutaneous disease. METHODS: Twelve patients with histologically confirmed cutaneous LE were given IVIg, with starting doses of 1 g/kgx2, followed by 400 mg/kg monthly until disease resolution or for 6 months. Disease assessment was by scoring erythema, induration, scaling and the extent before and at the end of therapy. Immunological parameters indicating systemic disease activity were measured before and after therapy. RESULTS: One patient became pregnant, five patients had complete or near complete clearing of their skin disease (>75%), two had partial but helpful improvement (>50%) and three had limited responses (<50%). One patient developed acute cutaneous vasculitis and received no further therapy. CONCLUSION: Overall, therapy was well tolerated and side effects were limited. A formal study of IVIg for cutaneous disease would be valuable, but evidence indicates that IVIg may be a useful therapy for cutaneous disease in lupus erythematosus. PMID- 14754651 TI - Inhibition of cold urticaria by desloratadine. AB - INTRODUCTION: The effect of 5 mg desloratadine for 4 days was tested with ice cubes before and after 4 days of treatment in 12 patients with cold urticaria. They had been asked not to take any antihistamine for at least 4 days. OBJECTIVE: To find the effect of desloratadine on wheals and itching induced by cold. METHOD: Ice-cubes in a thin plastic foil were placed on the inside of forearms for 0.5 to12 minutes. The time needed to produce a confluent urticaria, four or more small wheals and fewer than four small wheals were registered, together with the degree of itching. RESULTS: Before treatment a mean of 6.2 minutes was needed to cause a confluent urticaria, and itching occurred in seven of the patients. Four or more small wheals were observed after a mean of 3.7 minutes and fewer than four after a mean of 1.2 minutes. One 5 mg tablet of desloratadine was then given each morning for 4 days and the ice-cube test was repeated 4-5 hours after the last dose: no confluent urticaria and no itching occurred in any of the patients. Four or more small wheals were seen in two patients after 12 minutes, but there was no such reaction in the other 10 patients; in seven patients one to three small wheals developed. After 2-3 months of continued treatment four patients had noticed some wheals when out in the cold, but no itching. The symptoms of one patient with a delayed reaction after exposure to cold disappeared totally after a dose increase to 10 mg. CONCLUSION: Desloratadine markedly inhibited the reactions to cold induced urticaria. PMID- 14754652 TI - Comparative efficacy of cetirizine and fexofenadine in the treatment of chronic idiopathic urticaria. AB - OBJECTIVES: Antihistamines are the first line of therapy for chronic idiopathic urticaria (CIU). The present study was designed considering the lack of reports comparing the efficacy and safety of commonly prescribed cetirizine and fexofenadine in the treatment of CIU. METHODS: A total of 116 patients, aged 17 to 65 years, with CIU (urticarial wheals for at least two days per week for six consecutive weeks before entry) were enrolled in this randomised double-blind study. Study period was 28 days with patient visits on days 14 and 28 for investigator evaluation of the clinical response. Patient evaluation was on the basis of an analogue scale. Final response to treatment was judged as symptom free, partial improvement, and no improvement. RESULTS: Ninety-seven patients (52 cetirizine, 45 fexofenadine) completed the study. The response to treatment in both the groups at the end of treatment period was as follows; symptom free [cetirizine 27(51.9%), fexofenadine 2(4.4%)], partial improvement [cetirizine 19(36.5%), fexofenadine 19(42.2%)], no improvement [cetirizine 6(11.5%), fexofenadine 24(53.3%)]. Side effects noted were mild with no significant difference between the two. CONCLUSION: Cetirizine seems to have therapeutic advantage over fexofenadine in the treatment of CIU. PMID- 14754653 TI - An unusual late onset closed-comedone naevus: successful therapy with light cautery and a topical retinoid. PMID- 14754654 TI - Pemphigus foliaceus: an adverse reaction to lisinopril. AB - Lisinopril is an angiotensin converting enzyme inhibitor used in the treatment of cardiovascular disease. We report a case of pemphigus foliaceus in a 66-year-old male treated with lisinopril for hypertension and a previous myocardial infarction. The drug-induced variant of pemphigus is caused by a wide variety of drugs and is most frequently associated with captopril and penicillamine. It has not previously been reported in this commonly used drug. PMID- 14754655 TI - 0.1% Tacrolimus ointment in a patient with subacute cutaneous lupus erythematosus. PMID- 14754656 TI - A case of Parinaud's syndrome due to the remote effect of small cell carcinoma. AB - In a case with small cell carcinoma of lung, typical features of Parinaud's syndrome were seen. MRI and EEG revealed no involvement of central nervous system. Tests for specific infections and connective tissue diseases did not indicate abnormal findings. No electrophysiological findings suggesting the involvement of the peripheral and neuromuscular systems were seen using EMG-NCV measurements and repetitive electrical stimulations. Although no immunological studies were carried out, Parinaud's syndrome in this case can be attributed to a remote effect of the lung cancer. PMID- 14754657 TI - Serotonin modulates hypothalamic neuronal activity. AB - Effects of serotonin (5-HT) on electrophysiological activities of single hypothalamic arcuate neurons in rat brain slices were observed by extracellular recording. The results showed that (1) of 385 arcuate neurons observed, the patterns of spontaneous firing were divided into 3 categories: "slow irregular" (46.0%), "fast continuous" (22.6%), and "bursting" firing (31.4%); (2) of 149 neurons tested for 5-HT, most (55.0%) responded to the drug by decreasing firing rate, 22.2% by increasing firing rate, 11.4% exhibit biphasic pattern, and 11.4% did not respond to 5-HT application; (3) substitution of low Ca2-high Mg2+ artificial cerebrospinal fluid (ACSF) for normal ACSF did not abolish the 5-HT induced excitatory effect while it did abolish the 5-HT-induced inhibitory effect, suggesting the effect in latter cases was elicited by a Ca2+-dependent release of neuroactive substances; (4) cyproheptadine, a non-selective 5-HT receptor antagonist, antagonized the 5-HT-induced excitatory effect in all neurons tested, while pindolol, a 5-HT1A/1B receptor antagonist, antagonized the 5-HT-induced excitatory effect in 50% of neurons tested; (5) both cyproheptadine and pindolol antagonized the 5-HT-induced inhibitory effect; and (6) bicuculline, a GABAA receptor antagonist, antagonized the 5-HT-induced inhibitory effect. These results suggest that 5-HT may exert its excitatory effects directly through 5-HT1A/1B and other subtypes of 5-HT-receptors located on the surface of the neurons recorded, while its inhibitory effects are mediated indirectly through the activation of a local inhibitory GABAergic interneuron. PMID- 14754658 TI - Experience-dependent immediate early gene expression in the adult central nervous system: evidence from enriched-environment studies. AB - Here I discuss evidence from our group's work that implicates the immediate early genes NGFI-A and arc as possible regulators of neuronal plasticity. The enriched environment (EE) paradigm has been demonstrated to induce neural plasticity in both developing and mature mammals. Others and we have recently demonstrated that adult rats placed within an enriched environment underwent central nervous system wide increases in the expression levels for the IEGs NGFI-A and arc. The relationships between the altered expression profiles for both genes in response to an EE exposure, and their putative role in orchestrating network restructuring in response to enhanced environmental complexity are discussed. PMID- 14754659 TI - Dynamic lateralization: hostility, cardiovascular regulation, and tachistoscopic recognition. AB - This experiment tested a hypothesis linking the right cerebral regulation of hostility and cardiovascular arousal. It also replicates related research regarding hostility, cardiovascular regulation, and auditory recognition (Shenal Harrison, 2003) through the visual modality. Thirty low- and high-hostile participants (n = 30) were identified using the Cook Medley Hostility Scale (CMHS). Only right-handed male participants with no significant medical or psychological history completed the experiment. All participants completed the cold pressor paradigm. Cardiovascular measures (systolic blood pressure [SBP] and diastolic blood pressure [DBP]) were recorded and tachistoscopic lexical recognition procedures were administered before and after the physical stressor. The primary finding of this research was greater left cerebral activation (decreased cardiovascular reactivity) following the tachistoscopic lexical recognition tasks and greater right cerebral activation following the painful (cold pressor) stressor. PMID- 14754660 TI - Effects of L-arginine and NG-nitro L-arginine methyl ester on lipid peroxide, superoxide dismutase and nitrate levels after experimental sciatic nerve ischemia reperfusion in rats. AB - Nitric oxide (NO) has been reported to function in both cytoprotective and cytotoxic tissue ischemia-reperfusion (I/R). In this study, we evaluated the effects of L-arginine, the substrate for NO, and NG-nitro L-arginine methyl ester (L-NAME), NO synthase (NOS) inhibitor on super oxide dismutase (SOD) enzyme activity, malondialdehyde (MDA), a marker of lipid peroxidation, nitrate levels, and histopathological structure in rat sciatic nerve 2 h after ischemia, followed by 3 h of reperfusion. Reperfusion resulted in a significant increase in lipid peroxidation level and a decrease in nitrate level of the sciatic nerve. The increased level of lipid peroxidation was partly reduced by NOS inhibition. The decrease in sciatic nerve SOD level, observed in group subjected to I/R, was prevented by inhibition of NOS by L-NAME. These results were supported by histological findings that in the L-arginine-treated group, degenerations of both myelin sheath and axon were observed, while in the L- NAME-treated group, no pathological changes were detected. Our results suggested that excessive NO formation accelerates lipid peroxidation, as well as axonal degeneration on the early reperfusion period of the sciatic nerve. PMID- 14754661 TI - The effects of losartan and immobilization stress on heart rate variability and plasma corticosterone levels in rats. AB - In this study, the effects of losartan, an angiotensin II receptor antagonist, on heart rate variability and the changes of plasma corticosterone levels caused by immobilization stress were investigated. Losartan (3 mg/kg, p.o.) significantly prevented increases in plasma corticosterone levels in both losartan+acute stress and losartan+chronic stress groups. But, losartan did not prevent the diminution of the power of heart rate variability caused by stress. Our results supported the idea that the renin-angiotensin system is also involved in the stress- induced cardiovascular response, besides the autonomic nervous system. But, the effects of losartan on heart rate variability remained controversial. PMID- 14754662 TI - Effects of cigarette smoking on cognitive processing. AB - Several previous studies have reported that cigarette smoking enhances performance of cognitive processing. These enhancements are generally attributed to the pharmacological effects of nicotine, while there is some debate whether the effects of smoking/nicotine are a result of recovery from abstinence. Evoked potentials (EPs) and event related potentials (ERPs) of the brain have been applied as an index of information processing in a wide variety of normal and cognitive impaired subjects. This study was carried out on 20 healthy students (23 +/- 2.3 years old) from the medical faculty of City University. Study population comprised ten chronic cigarette smokers consuming an average of 14 +/- 4.2 cigarettes per day, with a history of smoking for more than one year. Ten non -smokers served as control. Standard oddball paradigm was presented, and EEG activity was recorded at the Fz, Cz, Pz electrode sites. Twenty responses to target stimuli were averaged at each location. N1, P2, N2, and P300 components were evaluated in these recordings. Amplitudes were measured relative to prestimulus baseline, and peak latencies were defined as the time point of maximum amplitude. It was found that there were no significant differences between either N1, P2, N2, P300 amplitudes or peak latency values of cigarette smokers and non smokers. As a result, chronic cigarette smoking generally does not improve cognitive processing. PMID- 14754663 TI - NMDA receptor subunits 2A and 2B decrease and lipid peroxidation increase in the hippocampus of streptozotocin-diabetic rats: effects of insulin and gliclazide treatments. AB - Recent studies indicate that diabetes mellitus changes N-methyl-D-aspartate (NMDA) receptor subunit composition and impairs cognitive functions. It also has been known that diabetes mellitus causes lipid peroxidation. This study examined the effects of streptozotocin-diabetes and insulin or gliclazide treatment on the hippocampal NMDA receptor subunit 2A and 2B (NR2A and NR2B) concentrations. In addition, malondial dehyde (MDA) levels were measured as a marker for lipid peroxidation. Eight weeks after the induction of diabetes MDA, levels were increased, and NR2A and NR2B concentrations were reduced. Insulin and gliclazide treatment partially prevented the reduction of NR2A and NR2B expression and prevented the elevation of MDA levels. There was no significant difference between the effects of insulin and gliclazide. The results suggest that the elevation of lipid peroxidation can be the primary biochemical disturbances in diabetes progression, and that changes in NMDA receptor subunit compositions can be involved in cognitive decline in diabetes. PMID- 14754664 TI - Emotion recognition deficits in the elderly. AB - In two studies, healthy elderly adults were poor at recognizing certain emotions. In study one, an emotion face morphed to express a new emotion. The elderly were impaired when recognizing anger and sadness, whereas no differences were found between the two age groups in recognizing fear or happiness, or in a task requiring reasoning about non=emotion stimuli. In study two, the elderly were impaired when judging which of two faces was more angry, sad, or fearful, but they were not impaired when judging other emotions or when judging which of two beakers was more full. The elderly were also impaired when matching emotion sounds to angry, sad, and disgusted faces, but not to other emotions and not when matching non-emotion (e.g., machine) sounds to machines. Elderly deficits were independent of performance on a task requiring basic face processing (gender recognition). Overall, the results provide support for an age-related decline in the recognition of some emotions that is independent of changes in perceptual abilities, processing speed, fluid IQ, basic face processing abilities, and reasoning about non face stimuli. Recognition of emotion stimuli might be mediated by regions of the brain that are independent from those associated with a more general cognitive decline. PMID- 14754666 TI - Fleeing the fire: transformation and gendered belonging in Thai HIV/AIDS support groups. AB - By the turn of the millennium, HIV had infected nearly one million people in Thailand. A large number of support groups now exist throughout the country. These groups have emerged as the primary forum through which having HIV is negotiated and normalized in Thai society. This is done by allowing members to publicly refashion their sense of self and its appropriate place in the world. However, the moral and social space created by support groups is not without its own structuring principles. The discursive strategies that shape support groups are embedded within local moral economies and frameworks of meaning. Gender and social identity are significant factors that influence the benefits to be gained from belonging. To date, women markedly outnumber men in most groups, and many members regard masculinity as a constraining factor on male participation. Within support groups, unwillingness to join is considered one reason for the perception that men with HIV seem to die sooner than do women with HIV. Clinically true or not, this belief has major ramifications. PMID- 14754667 TI - Lived food and judgments of taste at a time of disease. AB - Medical nutritional therapy (MNT) is a key feature of treatment for and management of type 2 diabetes. There are two elements to this therapeutic approach: collecting a diet history and prescribing therapeutic diets. For the clinical encounters observed in this paper, MNT often became a source of conflict between practitioners and patients. As clinicians endeavor to collect accurate information regarding food choices and eating so as to offer appropriate medical advice, and patients struggle to come to terms with a sickness in which food and eating have become toxic and risky, "judgments of taste" regarding food and patterns of eating become especially profound for both practitioners and those who seek treatment. Bourdieu's (1984) insights into the social situatedness of "taste" provides a useful framework for examining clinical practice and individual foodways. MNT is based upon and promotes a "taste for necessity," - "a form of adaptation to and consequently acceptance of the necessary" (ibid. 372), which judges food choices and eating patterns in terms of the bio-function of food and eating. In addition to this particular judgment of taste, study participants managing type 2 diabetes rely on other "judgments" that have been cultivated over the course of their own socially situated lives. At a time of disease these judgments of taste are conjoined as ongoing, multiply inflected lived histories of food and eating. Collecting life histories of food is one useful method for researching these patterned and idiosyncratic food and eating experiences. PMID- 14754668 TI - Delineating disease: self-management of leprosy identities in South India. AB - The national and international agencies working to eliminate leprosy are also dominant in setting the boundaries of official discourse on the issue. Within these boundaries the disease is commonly represented as a medical problem with negative social consequences, and it is believed that both problem and consequences will be resolved if leprosy is eliminated and its victims treated and (if necessary) reintegrated within their social groups. For those affected by leprosy the issues are frequently different, elimination in some respects representing a problem as much as a solution. Against this background, which I describe with reference to a group of leprosy-affected people in South India and their position vis- -vis leprosy organizations, I explore some of the contexts in which leprosy patients actively manage their own situations, often in defiance of prevailing development orthodoxies. I conclude that closer observation and analysis of the strategies patients use to manage their disease status have important policy implications. PMID- 14754669 TI - Living dangerously: driver distraction at high speed. AB - Recent research indicates that cell phone use can distract drivers from safe vehicle operation. However, estimates of the prevalence of cell phone use while driving have been limited to daytime hours and low-speed roadways. This paper describes the results of a study to estimate rates of cell phone use and other distractions by examining approximately 40,000 high-quality digital photographs of vehicles and drivers on the New Jersey Turnpike. The photographs, which originally were collected as part of a separate study, were taken both during the day and during the night and at different locations across the span of the Turnpike. A radar gun linked to the camera recorded the speeds of vehicles as they passed. This provided us with the speeds of every vehicle photographed, and allowed us to determine population counts of vehicles. A panel of three trained coders examined each photograph and recorded the presence of cell phone use by the drivers or any other distracting behavior. Demographic information on the driver was obtained during previous examinations of the photographs for an unrelated study. A rating was considered reliable when two out of the three coders agreed. Population estimates (and confidence intervals) of cell phone use and other distractions were estimated by weighting the cases by the inverse probability of vehicle selection. Logistic regression was used to predict cell phone use from demographic and situational factors. The results indicated that the most frequent distraction was cell phone use: 1.5% of the drivers on the Turnpike were using cell phones compared to the 3 to 4% use rates reported in the National Occupant Protection Use Survey (NOPUS) surveys conducted during the daytime on lower speed roadways. The Turnpike survey indicated that cell phones were used less on weekends and at night, and when the driver was exceeding the speed limit or had a passenger in the car. PMID- 14754671 TI - NCAP test improvements with pretensioners and load limiters. AB - New Car Assessment Program (NCAP) test scores, measured by the United States Department of Transportation's (USDOT) National Highway Traffic Safety Administration (NHTSA), were analyzed in order to assess the benefits of equipping safety belt systems with pretensioners and load limiters. Safety belt pretensioners retract the safety belt almost instantly in a crash to remove excess slack. They tie the occupant to the vehicle's deceleration early during the crash, reducing the peak load experienced by the occupant. Load limiters and other energy management systems allow safety belts to yield in a crash, preventing the shoulder belt from directing too much energy on the chest of the occupant. In NCAP tests, vehicles are crashed into a fixed barrier at 35 mph. During the test, instruments measure the accelerations of the head and chest, as well as the force on the legs of anthropomorphic dummies secured in the vehicle by safety belts. NCAP data from model year 1998 through 2001 cars and light trucks were examined. The combination of pretensioners and load limiters is estimated to reduce Head Injury Criterion (HIC) by 232, chest acceleration by an average of 6.6 g's, and chest deflection (displacement) by 10.6 mm, for drivers and right front passengers. The unit used to measure chest acceleration (g) is defined as a unit of force equal to the force exerted by gravity. All of these reductions are statistically significant. When looked at individually, pretensioners are more effective in reducing HIC scores for both drivers and right front passengers, as well as chest acceleration and chest deflection scores for drivers. Load limiters show greater reductions in chest acceleration and chest deflection scores for right front passengers. By contrast, in make-models for which neither load limiters nor pretensioners have been added, there is little change during 1998 to 2001 in HIC, chest acceleration, or chest deflection values in NCAP tests. PMID- 14754670 TI - The design of child restraint system (CRS) labels and warnings affects overall CRS usability. AB - A study was conducted that assessed the effectiveness of different child restraint system (CRS) label/warning designs on users' installation performance. Forty-eight paid participants installed a convertible CRS in a vehicle, and two child test dummies in a CRS, using one of four label conditions. The label conditions were: (1) no labels, (2) the manufacturer's labels that were already affixed to the CRS ("Current"), (3) labels that were designed according to a combination of the current U.S. regulations concerning CRS labels and recently proposed changes to these regulations ("Proposed"), and (4) labels that were designed according to human factors principles and guidelines, and that were based on a hierarchical behavioral task analysis ("Optimal"). Results demonstrated that, overall, the Optimal labels resulted in higher usability ratings and better task performance. This indicates that labels designed using human factors and task analyses that identify critical task information requirements for label features will result in increased user compliance with instructions, higher usability, and improved task performance. Surprisingly, having no labels on the CRS resulted in better installation performance than when either the Current or the Proposed label conditions were used. This indicates that label design can decrease task performance; the actual physical design of a CRS may be just as critical as label content in the installation choices provided to the user. Collectively, results suggest that implementation of the proposed changes to the U.S. regulations concerning CRS labeling would likely not result in increased performance or usability compared to existing manufacturer labels that follow the current guidelines. In order to achieve significantly better ease of-use and task performance, it would be necessary to implement features of the Optimal label condition. PMID- 14754672 TI - Older driver crash rates in relation to type and quantity of travel. AB - It is a well-established phenomenon that, notwithstanding their overall good crash record, older drivers have a higher than average rate of involvement in injury crashes when the rate is calculated by dividing crash numbers by distance driven. It has been hypothesised that at least some of this higher crash rate is an artefact of the different nature of driving undertaken by many older drivers. For example, driving in congested urban environments provides more opportunities for collisions than driving the same distance on a motorway. However, there have been few opportunities to investigate this theory, as relevant data are difficult to acquire. High-quality data from the New Zealand Travel Survey (1997/1998) were combined with crash data to enable a statistical model to estimate the risk of driver groups under various driving conditions characterised by the type of road used, time of day, day of week, and season of year. Despite elevated crash risks per distance driven compared with middle-aged drivers for most road types, older drivers were as safe as any other age group when driving on motorways. Accounting for the fragility of older drivers and their passengers in the risk estimates for other road types, older drivers appeared to have daytime risks comparable to 25 year-olds and night-time risks as low as any other age group. The driving patterns of older drivers (in terms of when and where they drive) were estimated to minimize their risks in comparison with the driving patterns of other age groups. These results are of interest to both policy makers and transportation planners working against the background of inevitable increases in the number of older drivers as the population ages. PMID- 14754673 TI - The effectiveness of ESP (Electronic Stability Program) in reducing real life accidents. AB - ESP (Electronic Stability Program) has recently been introduced onto the market in an effort to reduce the number and severity of loss-of-control automobile accidents. This reduction is expected to be particularly evident for accidents on roads with low friction (e.g., wet or icy conditions). This study aimed to evaluate the statistical effectiveness of ESP using data from accidents that occurred in Sweden during 2000 to 2002. To control for exposure, induced exposure methods were used, where ESP-sensitive to ESP-insensitive accidents and road conditions were matched in relation to cars equipped with and without ESP. Cars of similar, or in some cases identical, make and model were used to isolate the role of ESP. As predicted, the study showed a positive effect of ESP in circumstances where road surfaces have low friction. The overall effectiveness was 22.1 (+/-21) percent, while for accidents on wet roads, the effectiveness increased to 31.5 (+/-23.4) percent. On roads covered with ice and snow, the corresponding effectiveness was 38.2 (+/-26.1) percent. In addition, ESP was found to be effective for three different types of cars: small front-wheel drive; large front-wheel drive; and large rear-wheel drive. PMID- 14754674 TI - Potential risks of providing drinking drivers with BAC information. AB - The objective of this paper is to discuss the benefits and risks of providing drinkers with tools that allow them to estimate their blood alcohol concentration (BAC), and to examine the field usability of one commercially available tool. Drinking and driving laws are specified in terms of the driver's BAC, and there is concern that the absence of a method for drivers to accurately estimate their BAC level limits their ability to determine whether they can drive legally. A number of devices that provide a method for the individual to estimate or measure their BAC have been developed. Although some of these devices--such as the "know your limit" (KYL) cards--have been widely distributed, their effectiveness in encouraging good driving decisions have rarely been tested. This article describes a pilot study on the field usability of the Guardian Angel (GA) personal alcohol test in a field setting. The GA test analyzes saliva samples from drinking and indicates under which BAC category they fall (.00 -.04;.04 .08;.08+). The research examined whether drinkers could, in natural drinking environments, correctly administer and interpret the test results. The methodology involves sampling drinkers on a weekend night on and around the grounds of a large West Coast university as they traveled between off-campus parties, bars, and their dorms. They were asked to assess their own intoxication and impairment, then self-administer and interpret the Guardian Angel test. After interpreting the test, participants were asked to reassess their intoxication and impairment levels, and were given a breath BAC test using a calibrated unit. The results revealed that although the majority of drinkers' were able to administer the GA test, their interpretations did not correspond with actual BACs. The interpretations of the GA test produced false-negatives, underestimating actual BACs. Drinkers perceived themselves to be less intoxicated, on average, after interpreting the GA test results. In conclusion, this research addresses potential pros and cons of providing BAC information to drinkers. It underscores the importance of testing BAC estimation tools under field conditions and the potential risks associated with tests that do not produce accurate results. PMID- 14754675 TI - Rear impact responses of different sized adult Hybrid III dummies. AB - Rear impact sled tests were conducted using 5th, 50th, and 95th percentile Hybrid III dummies to evaluate proposed injury criteria. Different head restraint height (750, 800 mm) and backset (0, 50, 100 mm) positions were used to determine axial and shear forces, bending moments, and injury criteria (NIC, N(ij), and N(km)). The time sequence to attain each parameter was also determined. Three events were identified in the response. Event I was coincident with the maximum rearward motion of the torso, Event II occurred at the time of the peak upper neck flexion moment, and Event III occurred at the time of maximum rearward motion of the head. Parameters such as backset, head restraint height, seat-head restraint interaction, and anthropometry affected impact responses. Head rotations increased with increasing backset and increasing head restraint height. However, N(ij) and N(km) did not exhibit such clear trends. The 50th percentile dummy responded with consistent injury criteria values (e.g., the magnitude of the injury criteria increased with backset increase or head restraint height decrease). However, the 5th and 95th percentile dummies did not demonstrate such trends. These findings underscore the need to include subject anthropometry in addition to seat and head restraint characteristics for better assessment of rear impact responses. PMID- 14754676 TI - A comparison of injury criteria used in evaluating seats for whiplash protection. AB - A protocol has been proposed for testing seats for whiplash protection, however injury criteria have not yet been chosen. Assuming that whiplash symptoms arise from non-physiological motions of vertebral segments, we determined the ability of proposed criteria to predict peak individual vertebral displacements. Twenty eight volunteers were subjected to rear impacts while seated in a car seat with head restraint, mounted onto a sled. Accelerometers were used to record head and torso accelerations. The volunteer data was used as a basis for testing post mortem human specimens (PMHS). The seat was replaced by a platform onto which was mounted each of 11 cervico-thoracic spines. An instrumented headform was mounted to the upper end of the spine. The head restraint, head-to-restraint geometry, sled, and impact pulse remained the same. Head and T1 accelerations were measured and individual vertebral sagittal (XZ) plane rotations and translations were obtained from high speed video. Proposed injury criteria (NIC, Nkm, Nte, Nd) were tested for their ability to predict average, total, and peak intervertebral displacements. PMHS specimens had chest and head X (horizontal) and Z (vertical) linear accelerations similar to volunteers whose heads hit the head restraint. The best predictors were: Nd shear and peak intervertebral posterior translation (r(2) = 0.80), Nd extension and peak extension angle (r(2) = 0.70), and Nd distraction and peak distraction (r(2) = 0.51). Therefore consideration should be given to a displacement based injury criteria such as Nd in assessment of whiplash protection devices. PMID- 14754677 TI - Side impact crashes--factors affecting incidence and severity: review of the literature. AB - Many traffic crashes are side impact collisions resulting in significant death and injury. A review was conducted of the evidence of driver, road, and vehicle characteristics affecting either the risk of occurrence or the severity of injury in such crashes for papers published from 1996 to early 2003. For drivers, evidence was found of increased crash risk or injury severity only for age and age-related medical conditions (e.g., dementia). Traffic roundabouts and other traffic control devices--stop signs, traffic lights, and so on--had mixed results; traffic controls were better than no controls, but their effectiveness varied with circumstance. Most vehicle characteristics have had little or no effect on crash occurrence. Antilock braking systems (ABS) in the striking vehicle had been anticipated to reduce the risk of crashes, but so far have demonstrated little effect. The primary emphasis in vehicle design has been on protective devices to reduce the severity of injury. Disparity in the size of the two vehicles, especially when the struck vehicle is smaller and lighter, is almost a consistent risk factor for occupant injury. The occupants of light trucks, however, when struck by passengers cars on the opposite side, were at higher risk of injury. Wearing seat belts had a consistently protective effect; airbags did not, but there were few studies, and no field studies, of lateral airbags found. Of all the characteristics examined, vehicle design, including occupant restraints, is the most easily modified in the short term, although road design, traffic control, and the monitoring of older drivers may also prove effective in reducing side impact crashes in the longer term. PMID- 14754678 TI - Evaluation of pedestrian subsystem test method using legform and upper legform impactors for assessment of high-bumper vehicle aggressiveness. AB - In accidents involving sports utility vehicles (SUVs), injuries to pedestrian leg, knee ligaments, and femur are likely to occur. Therefore, the European Enhanced Vehicle Safety Committee proposed two subsystem test methods for evaluation of SUV bumper aggressiveness. Such evaluation can be conducted by means of either a legform impactor (evaluation of risk of knee and tibia injury), or an upper legform impactor (evaluation of risk of thigh and pelvis injury) test. Each of these two test methods has its own injury criteria and injury acceptance levels. Therefore, the first objective of this research is to clarify any differences between the test results obtained when evaluating SUV bumper aggressiveness by means of these two impactors. The second objective is to determine whether or not a legform impactor can be applied to estimate the risk of femur fracture, and if an upper legform impactor can be used to estimate the risk of knee ligament injury. The present results indicate the test method using an upper legform impactor yields higher ratios of injury criteria to the relevant EEVC/WG17 injury acceptance levels than by using a legform impactor. Thus, the upper legform impactor test rates an SUV bumper as more aggressive than the legform impactor test. The present study suggests the lower leg acceleration obtained by the legform impactor can be used to adequately assess the risk of femur fracture, when evaluating the aggressiveness of an SUV bumper using proposed injury acceptance levels reported in the literature. Similarly, the impact force obtained by the upper legform impactor can be used to assess the risk of cruciate ligament injury. PMID- 14754679 TI - Outcome of motorcycle riders at one year post-injury. AB - OBJECTIVE: There is little information about the long-term effects of injury on victims of motorcycle crashes. Following the repeal of the mandatory helmet law in Florida, we studied the impact of crashes on riders and their outcomes one year post-injury. SUBJECTS: All patients involved in a motorcycle crash over a six-month time period were assessed at one year post-injury. Demographic information, health status, motorcycle and helmet usage, and employability were surveyed. RESULTS: There were 94 patients evaluated at our center for the first six-month period following the repeal of the helmet law, where 50 (56%) were wearing a helmet at the time of injury. Of the 94, we were able to contact 45 (48%) at one-year follow up. Fourteen (31%) of those contacted were not wearing a helmet at the time of their crash. At one year post-injury, 23 (51%) reported physical deficits, while 39 (86%) were working. Only 12 (27%) were riding a motorcycle again, and 11 (92%) were wearing a helmet. CONCLUSIONS: Motorcycle riders at one year post-injury continue to experience physical deficits related to their accidents. There is a small percentage of riders that resume riding a motorcycle after injury. Being involved in a motorcycle accident has deterred the injured rider from riding a motorcycle again at one year post-injury. PMID- 14754680 TI - Letter to the editors and reply on: "Air bag effectiveness as a function of impact speed" Traffic Injury Prevention, 4:128-135, 2003. PMID- 14754686 TI - Obesity: a sign of the times. PMID- 14754687 TI - Maternal morbid obesity and the risk of adverse pregnancy outcome. AB - OBJECTIVE: To evaluate whether morbidly obese women have an increased risk of pregnancy complications and adverse perinatal outcomes. METHODS: In a prospective population-based cohort study, 3,480 women with morbid obesity, defined as a body mass index (BMI) more than 40, and 12,698 women with a BMI between 35.1 and 40 were compared with normal-weight women (BMI 19.8-26). The perinatal outcome of singletons born to women without insulin-dependent diabetes mellitus was evaluated after suitable adjustments. RESULTS: In the group of morbidly obese mothers (BMI greater than 40) as compared with the normal-weight mothers, there was an increased risk of the following outcomes (adjusted odds ratio; 95% confidence interval): preeclampsia (4.82; 4.04, 5.74), antepartum stillbirth (2.79; 1.94, 4.02), cesarean delivery (2.69; 2.49, 2.90), instrumental delivery (1.34; 1.16, 1.56), shoulder dystocia (3.14; 1.86, 5.31), meconium aspiration (2.85; 1.60, 5.07), fetal distress (2.52; 2.12, 2.99), early neonatal death (3.41; 2.07, 5.63), and large-for-gestational age (3.82; 3.50, 4.16). The associations were similar for women with BMIs between 35.1 and 40 but to a lesser degree. CONCLUSION: Maternal morbid obesity in early pregnancy is strongly associated with a number of pregnancy complications and perinatal conditions. LEVEL OF EVIDENCE: II-2 PMID- 14754688 TI - Intrauterine pressure during the second stage of labor in obese women. AB - OBJECTIVE: The perception that obese women have longer labors and a higher frequency of operative delivery because they are "poor pushers" persists despite the absence of objective study. We tested the hypothesis that obese women generate inadequate intrauterine pressure during the second stage of labor. METHODS: Intrauterine pressure was prospectively measured in 71 women during the second stage of labor. Obesity was defined as a body mass index (BMI) greater than 29 (n = 17). A BMI below 25 was normal (n = 40). Women with a BMI between 26 and 29 (n = 14) were considered overweight. All women labored with epidural analgesia and were alert and responsive throughout the study. After recording the baseline contractility, a standardized Valsalva maneuver was performed during contractions. The area under the intrauterine pressure curve (integral) was used as an estimate of uterine contractility. RESULTS: All women delivered vaginally. There were no significant differences in baseline uterine contractility among obese, overweight, and normal women either before (obese 1,787 mm Hg/s; 95% confidence interval [CI] 1,164, 2,742 versus normal 1,569 mm Hg/s; 95% CI 718, 2,371 versus overweight 1,770 mm Hg/s; 95% CI 1,305, 2,835; P =.223) or during Valsalva maneuver (obese 2,831 mm Hg/s; 95% CI 1,771, 4,599 versus normal 2,637 mm Hg/s; 95% CI 1,240, 4,390 versus overweight 2,813 mm Hg/s; 95% CI 1,209, 4,982; P =.742). A BMI greater than 25 was associated with a higher frequency of oxytocin augmentation (P =.037). Univariate analysis revealed a relationship between labor duration and BMI (r = 0.299, P =.018). Obese women labored longer during the active phase (one-way analysis of variance, P =.02), but second-stage duration was similar among groups (one-way analysis of variance P =.44). Obesity did not increase the incidence of perineal lacerations (P =.82) or frequency of operative delivery (relative risk obese versus nonobese = 0.212; 95% CI 0.04, 1.05). CONCLUSION: Obese women produce second-stage intrauterine pressures equivalent to women with a normal BMI, although they may require oxytocin augmentation more often. LEVEL OF EVIDENCE: II-2 PMID- 14754689 TI - Correlates of weight loss behaviors among low-income African-American, Caucasian, and Latina women. AB - OBJECTIVE: To examine the prevalence and correlates of weight reduction behaviors among low-income women. METHODS: A total of 1,709 Caucasian, African-American, and Latina women aged 12-58 years attending a federally funded family planning clinic reported their weight loss behaviors during the past 30 days, including using diet pills, exercising, purging (vomiting, laxatives, diuretics), and dieting. Approximately 60% (n = 999) had a body mass index of 25 or greater. RESULTS: Overall, 35.3% (n = 603) of women dieted, 43.7% (n = 746) exercised, 15.1% (n = 258) used diet pills, and 4.3% (n = 69) purged. Only 14.8% (n = 253) of the sample reported both dieting and exercising. The odds of exercising for weight loss decreased as parity increased. Those who smoked currently or in the past were more likely than nonsmokers to report purging (odds ratio [OR] 2.5; 95% confidence interval [CI] 1.2, 5.4). African Americans were least likely to diet or exercise compared with Caucasians and Latinas. Exposure to family members using diet pills (OR 4.6; 95% CI 3.2, 6.5), dieting (OR 2.1; 95% CI 1.6, 2.8), or purging (OR 5.6; 95% CI 2.7, 11.9) was associated with increased odds of performing these behaviors oneself. CONCLUSION: This research demonstrates that low-income women frequently use maladaptive strategies, such as diet pills and purging, to lose weight. Obesity and family exposure to these behaviors places women at increased risk of unhealthy behaviors. Interventions designed to reduce obesity must include precautions regarding the dangers of these practices. LEVEL OF EVIDENCE: III PMID- 14754690 TI - Diabetic pregnancy associated with increased epidermal growth factor in cord serum at term. AB - OBJECTIVE: Epidermal growth factor is a ubiquitous mitogen that also possesses insulin-like properties. Fetal mal-growth is associated with altered epidermal growth factor levels. Maternal diabetes is frequently complicated by macrosomia, but the effect of maternal diabetes on fetal epidermal growth factor levels is not known. We studied cord serum epidermal growth factor concentrations in pregnancies complicated by diabetes and in normal pregnancies. METHODS: Cord serum epidermal growth factor concentrations were measured at birth by a sandwich type time-resolved immunofluorometric assay in 63 pregnancies complicated by insulin-dependent diabetes mellitus, in 25 pregnancies complicated by insulin treated gestational diabetes, and in 56 normal pregnancies. RESULTS: Cord serum epidermal growth factor correlated positively with the duration of pregnancy in diabetic and normal pregnancies. In a subgroup of women at similar gestational ages (38-39 weeks), cord serum epidermal growth factor concentrations were higher in pregnancies complicated by insulin-dependent diabetes mellitus (962 +/- 211 ng/L, P =.047; n = 9) and in pregnancies complicated by gestational diabetes (1133 +/- 115 ng/L, P =.001; n = 9) than in controls (564 +/- 75 ng/L; n = 22). In multiple regression analysis, only umbilical artery hemoglobin in diabetic pregnancies and vaginal delivery in normal pregnancies were associated with cord serum epidermal growth factor. CONCLUSION: Epidermal growth factor concentrations are higher than normal in fetuses of diabetic mothers at term. Pregnancy complications, such as hypertensive disorders, fetal hypoxia and fetal malgrowth, may not explain the rise in epidermal growth factor levels. We hypothesize that the rise in epidermal growth factor levels is a metabolic response of the fetoplacental unit to diabetes-related hyperglycemia. LEVEL OF EVIDENCE: III PMID- 14754691 TI - Relief of hot flushes with new plant-derived 10-component synthetic conjugated estrogens. AB - OBJECTIVE: A study was conducted to evaluate the safety and efficacy of 3 different doses of synthetic conjugated estrogens B, a new plant-derived 10 component conjugated estrogens product, for the treatment of menopausal vasomotor symptoms. METHODS: This was a randomized, double-blind, placebo-controlled trial. Highly symptomatic menopausal women (N = 281) received 12 weeks of a once-daily oral treatment with 0.3 mg, 0.625 mg, or 1.25 mg of 10-component synthetic conjugated estrogen or placebo. Patients recorded the daily frequency and severity of hot flushes. Statistical analyses compared results at weeks 4, 8, and 12 with baseline values. RESULTS: Statistically significant reductions (P <.05) in the frequency and severity of vasomotor symptoms were observed for all 3 dosage strengths of 10-component synthetic conjugated estrogen compared with placebo. The most commonly reported adverse events in all treatment groups were headaches. No difference in the incidence of treatment-related adverse events was seen between placebo and 10-component synthetic conjugated estrogen groups. CONCLUSION: The 0.3-mg, 0.625-mg and 1.25-mg dose strengths of 10-component synthetic conjugated estrogen significantly reduced the frequency and severity of vasomotor symptoms compared with placebo, and were well tolerated during this 12 week study. LEVEL OF EVIDENCE: I PMID- 14754692 TI - Postmenopausal hormone therapy and risk of developing urinary incontinence. AB - OBJECTIVE: To better understand associations between post-menopausal hormone therapy and the development of urinary incontinence. METHODS: Postmenopausal hormone use was assessed via biennial mailed questionnaires beginning in 1976 among Nurses' Health Study participants. In 1996, 39,436 post-menopausal women aged 50-75 years reported no leaking of urine and were followed-up for 4 years to identify incident cases of incontinence. We used logistic regression to estimate multivariable-adjusted relative risks (RRs) and 95% confidence intervals (CIs) for the relation of postmenopausal hormone use from 1976 to 1996 to the development of incontinence from 1996 to 2000. RESULTS: We identified 5,060 incident cases of occasional (leaking urine 1-3 times/month) and 2,495 cases of frequent incontinence (leaking at least weekly) for average yearly incidence rates of 3.2% and 1.6%, respectively. The risk of incontinence was elevated among women taking postmenopausal hormones compared with women who had never taken hormones (oral estrogen: RR 1.54, 95% CI 1.44, 1.65; transdermal estrogen: RR 1.68, 95% CI 1.41, 2.00; oral estrogen with progestin: RR 1.34, 95% CI 1.24, 1.44; transdermal estrogen with progestin: RR 1.46, 95% CI 1.16, 1.84). There was little risk after the cessation of hormones (RR 1.14, 95% CI 1.06, 1.23) and a decreasing risk of incontinence with increasing time since last hormone use; 10 years after stopping hormones, the risk was identical in women who had and had never taken hormone therapy (RR 1.02, 95% CI 0.91, 1.14). CONCLUSION: Postmenopausal hormone therapy appears to increase risk of developing urinary incontinence. This risk does not vary by route of administration, type of hormones, or dose taken, but is diminished upon cessation of use. LEVEL OF EVIDENCE: II-2 PMID- 14754694 TI - Transition from estrogen-progestin to raloxifene in postmenopausal women: effect on vasomotor symptoms. AB - OBJECTIVE: To compare vasomotor symptoms after transition from estrogen-progestin therapy to raloxifene 60 mg/d with and without a placebo washout. METHODS: Postmenopausal women currently taking continuous combined estrogen-progestin therapy (conjugated equine estrogen, 0.625 mg/medroxyprogesterone acetate, 2.5 or 5 mg) daily for 5 or more months were enrolled. Women were randomized to 1 of 4 blinded regimens: 1) 12 weeks estrogen-progestin; 2) 12 weeks placebo; 3) 4 weeks placebo, then 8 weeks raloxifene; or 4) 12 weeks raloxifene. For the final 36 weeks, all subjects received raloxifene. Vasomotor symptoms were assessed by patient diaries. RESULTS: A total of 266 women (mean age 57.5) were enrolled. Mean hot flush frequency at baseline was approximately 1 per week in the entire population, with 16% of women reporting hot flushes. Mean frequency and severity of hot flushes during the first 12 weeks of the study were statistically greater in the 3 groups transitioned off estrogen-progestin (range of hot flushes per week: 4 weeks, 11-12; 8 weeks, 18-24; 12 weeks, 13-16), as compared with those continuing estrogen-progestin, with no difference between these 3 groups (P> or =.1). Approximately 50-70% of these women reported hot flushes, generally rated as mild to moderate by participants, after estrogen-progestin discontinuation. CONCLUSION: A large proportion of women discontinuing estrogen-progestin experience hot flushes. Raloxifene does not appear to increase the frequency or severity of vasomotor symptoms in women discontinuing estrogen-progestin more than that observed with placebo treatment after estrogen-progestin discontinuation. Transition from estrogen-progestin to raloxifene with no washout period therefore may be acceptable. LEVEL OF EVIDENCE: I PMID- 14754693 TI - Effect of raloxifene on urinary incontinence: a randomized controlled trial. AB - OBJECTIVE: To estimate the effect of 3 years of treatment with raloxifene on urinary incontinence in postmenopausal women. METHODS: We used measures of urinary incontinence severity, frequency, and type in the Multiple Outcomes of Raloxifene trial, a multicenter randomized, controlled trial of women who were at least 2 years postmenopausal with osteoporosis. At 10 U.S. sites of this trial, 963 women randomly assigned to raloxifene or placebo completed questionnaires about incontinence at baseline and 3 years later. We analyzed the odds of worsening severity and frequency of incontinence and type of incontinence after 3 years of treatment with raloxifene. RESULTS: The mean age of our subjects was 68.3 +/- 7 years. After 3 years of treatment, there was no significant difference between raloxifene and placebo groups in urinary incontinence severity (multivariable odds ratio [OR] 1.02; 95% [CI] 0.78, 1.34). The majority of the women (60%) had no change in urinary incontinence episodes from baseline to year 3. The odds of worsening urinary incontinence severity after 3 years of raloxifene treatment were 1.05 (95% CI 0.75, 1.48). Similarly, the odds of developing new onset incontinence were 0.95 (95% CI 0.59, 1.52). Finally, raloxifene did not effect the odds of having stress (OR 1.01; 95% CI 0.71, 1.43) or urge (OR 1.20; 95% CI 0.86, 1.68) incontinence after 3 years of use. CONCLUSION: In postmenopausal women with osteoporosis, 3 years of treatment with raloxifene had no effect on urinary incontinence. LEVEL OF EVIDENCE: I PMID- 14754695 TI - Single versus weekly courses of antenatal corticosteroids in preterm premature rupture of membranes. AB - OBJECTIVE: This study was performed to evaluate the efficacy of weekly courses of antenatal corticosteroids compared with a single course in women with preterm premature rupture of membranes (PROM). METHODS: A planned secondary analysis of women with preterm PROM who participated in a multicenter, randomized trial of weekly courses of antenatal corticosteroids versus single-course therapy was performed. After their first course of standard antenatal steroid therapy, administered between 24 to 32-6/7 weeks of gestation, consenting women were randomly assigned to receive betamethasone versus placebo injections weekly until 34-0/7 weeks of gestation. Maternal and neonatal morbidities were compared between the 2 groups. RESULTS: Of the 161 women with preterm PROM, 81 women were assigned to receive weekly courses of steroids and 80 to the single-course group. There were no significant differences in composite morbidity between the groups (27 [34.2%] of 81 patients versus 33 [41.8%] of 80 patients, P =.41). Chorioamnionitis was higher in patients who received weekly courses of antenatal steroids (39 [49.4%] of 81 patients versus 25 [31.7%] of 80 patients, P =.04). CONCLUSION: Weekly courses of antenatal steroids in women with preterm PROM did not improve neonatal outcomes beyond that achieved with single-course therapy and was associated with an increased risk of chorioamnionitis. Antenatal steroid therapy should not be routinely repeated in patients with preterm PROM. LEVEL OF EVIDENCE: I PMID- 14754696 TI - Predicting failed trial of labor after primary cesarean delivery. AB - OBJECTIVE: To apply published scoring systems retrospectively to patients who had undergone a trial of labor after cesarean delivery to estimate whether there was a score at which a trial of labor should be discouraged. METHODS: Patients with 1 previous cesarean delivery who then delivered between January 1, 1998, and December 31, 1998, were studied. An investigator blinded to outcome assigned scores using 3 different scoring systems. Student t test, chi(2), analysis of variance, and receiver operating curve analysis were performed. P <.05 was significant. RESULTS: Seventy-six percent (117/153) of trial of labor patients had a vaginal birth after cesarean delivery. Successful vaginal birth after cesarean delivery patients had significantly different mean scores using all 3 scoring systems, but none of the systems accurately predicted failed trial of labor resulting in cesarean delivery. Unfavorable scores were associated with high rates of major complications. CONCLUSIONS: An unfavorable score predicting a high rate of complications and more failed trials of labor may help in counseling patients considering trial of labor. A better system to predict the success or failure of trial of labor is needed. LEVEL OF EVIDENCE: III PMID- 14754697 TI - Maternal temperature monitoring during labor: concordance and variability among monitoring sites. AB - OBJECTIVE: Elevated maternal temperature in labor is associated with adverse immediate and long-term neonatal outcomes. Conventional methods of temperature measurement may not reflect the intrauterine temperature, which constitutes the fetal environment. The purpose of this study was to ascertain the most reliable noninvasive method of temperature monitoring in labor that would best reflect changes in intrauterine temperature. METHODS: Women in labor receiving epidural analgesia had temperature readings taken every 10 seconds from the uterine cavity, ear canal, and skin surface of the leg and abdomen and hourly from the mouth. RESULTS: Eighteen patients were studied for a mean of 228 minutes (range 56-464 minutes). The best indicator of intrauterine temperature was oral temperature, with a mean intraclass correlation coefficient of 0.6 (95% confidence interval 0.42, 0.77). On average, oral temperature underestimated intrauterine temperature by 0.8 degrees C (95% confidence interval 0.7 degrees C, 1 degrees C). Allowing for this, oral temperature greater than 37.2 degrees C detected an intrauterine temperature greater than 38 degrees C with a sensitivity of 81% and a specificity of 96%. The intraclass correlation coefficients of all other sites with intrauterine temperature were poor (0.1 or less). As expected, the temperature at all sites increased as labor progressed. CONCLUSION: Oral temperature, measured carefully, has an acceptable correlation with intrauterine temperature and is recommended for routine detection of maternal pyrexia in labor. Continuous skin and external auditory canal temperature measurements did not correlate well. LEVEL OF EVIDENCE: II-3 PMID- 14754698 TI - Cerebrovascular hemodynamics in pregnant women with mild chronic hypertension. AB - OBJECTIVE: To evaluate and compare the cerebrovascular autoregulation in pregnant normotensive and mild chronic hypertensive patients without preeclampsia. METHODS: Transcranial Doppler ultrasound was used to measure peak, end-diastolic, and mean velocities in the middle cerebral arteries of 34 normotensive and 17 mild chronic hypertensive women in the third trimester of pregnancy. Measurements were performed in the left lateral position at baseline, during 5% CO(2) inhalation, and during an isometric handgrip test. Mean pulsatility index, resistance index, and cerebral perfusion pressure at each time were compared using 2-way repeated measures analysis of variance. Using an alpha error of 5%, the statistical power to identify differences in middle cerebral artery indices in response to the two maneuvers was at least 90% and 50% in comparison between the two groups. Significance was P <.05. RESULTS: Pregnant women with mild chronic hypertension had higher baseline mean blood pressure but similar pulsatility index (0.73 versus 0.75), resistance index (0.50 versus 0.50), and cerebral perfusion pressure (59.9 versus 61.8 mm Hg) compared with normotensive pregnant women. Both maneuvers caused a significant reduction in pulsatility index and resistance index and higher cerebral perfusion pressure. No significant differences were noted in the response to either 5% CO(2) inhalation or isometric handgrip test between the two groups. CONCLUSION: Pregnant women with mild chronic hypertension show normal cerebral vasomotor reactivity to CO(2) breathing and isometric handgrip. This suggests that the abnormal cerebrovascular autoregulation in preeclampsia is not directly linked to the elevated blood pressure but rather is determined by a separate pathophysiologic pathway. LEVEL OF EVIDENCE: II-2 PMID- 14754699 TI - Screening for partner violence: direct questioning or self-report? AB - OBJECTIVE: To compare the effectiveness of a self-report checklist with a standard set of direct questions in identifying women who are experiencing domestic partner violence. METHODS: Medical records were reviewed for evidence of positive partner violence for women attending the Royal Brisbane and Women's Hospital prenatal clinic between August and September 2002. RESULTS: Records (n = 1,596) were audited, and 937 (58.7%) contained both forms. The self-report check list identified a greater number of "cases" of partner violence (151) than the direct questions (66), with the level of agreement between the two instruments being only "fair" (Kappa coefficient.34). Each of the methods identified 7 cases of major abuse, which would have been missed if only 1 instrument had been used. All cases where women stated that they were afraid of their partner using the direct questions were also identified using the self-report checklist. CONCLUSION: A self-report checklist is an effective alternative to direct questioning in detecting women who are experiencing partner violence and is acceptable to women. LEVEL OF EVIDENCE: II-3 PMID- 14754700 TI - Interim guidance for the use of human papillomavirus DNA testing as an adjunct to cervical cytology for screening. AB - Human papillomavirus (HPV) DNA testing was recently approved by the Food and Drug Administration for use as an adjunct to cytology for cervical cancer screening. To help provide guidance to clinicians and patients when using HPV DNA testing as an adjunct to cervical cytology for screening, a workshop was cosponsored by the National Institutes of Health-National Cancer Institute, American Society of Colposcopy and Cervical Pathology (ASCCP), and American Cancer Society. Consensus was reached based on a literature review, expert opinion, and unpublished results from large ongoing screening studies. The conclusions of the workshop were that HPV DNA testing may be added to cervical cytology for screening in women aged 30 years or more. Women whose results are negative by both HPV DNA testing and cytology should not be rescreened before 3 years. Women whose results are negative by cytology, but are high-risk HPV DNA positive, are at a relatively low risk of having high-grade cervical neoplasia, and colposcopy should not be performed routinely in this setting. Instead, HPV DNA testing along with cervical cytology should be repeated in these women at 6 to 12 months. If test results of either are abnormal, colposcopy should then be performed. This guidance should assist clinicians in utilizing HPV DNA testing in an effective manner, while minimizing unnecessary evaluations and treatments. PMID- 14754701 TI - Effectiveness of screening for cervical cancer in an inpatient hospital setting. AB - OBJECTIVE: To estimate the effectiveness of an inpatient, hospital-based cervical cancer screening program at a single institution. METHODS: Between January 1999 and December 2002, 1,117 women admitted to the Johns Hopkins Hospital underwent Papanicolaou (Pap) test screening during their hospital stay. In that time period, 111,933 women were screened at all of the combined Hopkins outpatient clinics. We compared rates of abnormal Pap tests in these cohorts (retrospective cohort study). Our main outcome measure was the prevalence of abnormal Pap tests among the screening population by age group, ethnicity, and insurance status compared between our outpatient and inpatient populations. RESULTS: The prevalence of abnormal Pap tests in the inpatient cohort was twice as high as that in the outpatient setting (15.5% versus 7%). The prevalence of high-grade squamous intraepithelial lesions (HSIL), the immediate precursor lesion to cervical cancer, was nearly 5-fold higher in the inpatient cohort compared with the outpatient cohort (3% versus 0.7%). In multivariable models, younger women had greater risk for all types of abnormal Pap tests, and black women had greater risk for HSIL. Previous abnormal Pap and human immunodeficiency virus-positive status were associated with all abnormal tests and with HSIL results. CONCLUSIONS: A hospital-based, inpatient Pap test program is an efficient strategy for targeting limited screening funds toward women at high risk of invasive cervical cancer. LEVEL OF EVIDENCE: II-2 PMID- 14754702 TI - ZYC101a for treatment of high-grade cervical intraepithelial neoplasia: a randomized controlled trial. AB - OBJECTIVE: The objective of this study was to assess the safety and efficacy of a novel therapeutic, ZYC101a, for the treatment of women with histologically confirmed cervical intraepithelial neoplasia (CIN) 2/3. ZYC101a contains plasmid DNA-encoding fragments derived from the E6 and E7 proteins of human papillomavirus (HPV) 16 and 18, and is formulated within small biodegradable microparticles. METHODS: A multicenter, double-blind, randomized, placebo controlled trial was conducted in a group of women with biopsy-confirmed CIN 2/3. Subjects were randomized to 3 intramuscular doses of either placebo or ZYC101a (100 or 200 microg). Six months after the first injection, subjects underwent cervical conization. The primary endpoint for this study was histologically confirmed resolution of CIN 2/3. A total of 161 subjects were randomized, dosed, and evaluated for safety. After central pathology review, 127 subjects were evaluable for efficacy. RESULTS: The most common adverse events were related to the injection site, were mild to moderate, and did not worsen at later treatments. The proportion of subjects who resolved was higher in the ZYC101a groups compared to placebo (43% versus 27%), but the difference was not statistically significant (P =.12). In a prospectively defined population of women younger than 25 years (n = 43), resolution was significantly higher in the combined ZYC101a groups compared to placebo (70% versus 23%; P =.007). ZYC101a activity was not restricted to HPV-16-or HPV-18-positive lesions. CONCLUSIONS: ZYC101a was shown to be well tolerated in all patients and to promote the resolution of CIN 2/3 in women younger than 25 years. LEVEL OF EVIDENCE: I PMID- 14754704 TI - Fetal ultrasound training for obstetrics and gynecology residents. AB - OBJECTIVE: To assess the present state of fetal ultrasound training in the United States from the perspective of obstetrics and gynecology ultrasound program directors and residents. METHODS: One hundred thirty-six ultrasound program directors from 254 accredited obstetrics and gynecology residency programs completed a web-based survey regarding obstetric ultrasound training for residents. Questions were presented in yes-or-no, ranking, short-answer, and open comment formats that examined general teaching environment and curriculum content. These results were compared with a mandatory fetal ultrasound training survey that was independently administered to 4,666 obstetrics and gynecology residents during the 2003 Council on Resident Education in Obstetrics and Gynecology (CREOG) In-Training Examination. Friedman one-way analysis of variance was used to compare ranked nonparametric data with the Dunn posttest. Statistical significance was taken at the P <.05 level. RESULTS: Fifty-four percent of accredited obstetrics and gynecology residencies responded to the survey of ultrasound directors from November 2000 to April 2003. Nearly all responding directors were obstetrician-gynecologists, many of whom had subspecialty training in maternal-fetal medicine. Full-time faculty and sonographers were the most important individuals contributing to ultrasound training for obstetrics and gynecology residents. Hands-on scanning and observation were the most significant educational activities for ultrasound training. Ultrasound program directors generally rated the overall preparedness of residents as ranging from adequate to excellent. The most important learning obstacles were limited curriculum and faculty time. Most programs evaluated competency by direct observation of scanning skills. According to the CREOG survey, only 16.3% of residents indicated that the performance and interpretation of fetal ultrasound examinations were mandatory program requirements. Nearly two thirds of residents believed that their training would be adequate by the time of graduation. Only 18.4% of residents, however, were planning to perform or interpret fetal ultrasound scans in clinical practice. CONCLUSION: Fetal ultrasound training for obstetrics and gynecology residents is perceived by most ultrasound program directors and residents to be adequate. Future development of standardized guidelines and competency assessment tools should consider that approximately one fifth of obstetrics and gynecology residents are currently planning to use this diagnostic modality in clinical practice. LEVEL OF EVIDENCE: II-2 PMID- 14754703 TI - The association between fetal sex and preterm birth in twin pregnancies. AB - OBJECTIVE: To assess the association between the fetal sex and preterm birth. METHODS: We performed a retrospective population-based cohort study using the 1995-1997 registration twin data in the United States (148,234 live-birth twin pairs). The twin pairs were divided into 3 groups: male-male (male-male), female female, and opposite sex. We used 3 different cutoff values of preterm birth: less than 28, 32, and 36 gestational weeks. The preterm birth rates among the 3 study groups were compared, and the adjusted risk ratios (relative risk) were estimated by multiple logistic regression. RESULTS: The male-male twin pairs had the highest pre-term birth rate (less than 28 weeks: 4.9%; less than 32 weeks: 12.4%; less than 36 weeks: 40.2%), the female-female twin pairs were intermediate (less than 28 weeks: 4.1%; less than 32 weeks: 10.6%; less than 36 weeks: 37.8%), and the opposite-sex twin pairs had the lowest rate (less than 28 weeks: 4.1%; less than 32 weeks: 10.1%; less than 36 weeks: 36.8%). Adjustment for important confounding factors or excluding twin pairs born to mothers who had an induction of labor or a cesarean delivery with medical complications did not change the results. The adjusted relative risks (95% confidence intervals) were 1.19 (1.11, 1.27), 1.21 (1.16, 1.26), and 1.09 (1.07, 1.11), respectively, for male-male twins compared with the opposite-sex twins under the 3 different cutoff values of preterm births. CONCLUSION: Male sex is associated with increased risk of preterm births in twin pregnancy. LEVEL OF EVIDENCE: II-2 PMID- 14754705 TI - Increasing recruitment of quality students to obstetrics and gynecology: impact of a structured clerkship. AB - OBJECTIVE: To evaluate the satisfaction of the third-year medical students with their basic third-year obstetrics and gynecology rotation before and after a concerted effort was made to improve the rotation with prompt feedback and problem-based case learning. METHODS: At the end of each rotation, third-year medical students were asked to anonymously fill out an "in-house" course evaluation. A 1-5 rating scale to evaluate quality of teaching, instructor's commitment and enthusiasm, and enhancement of professional development was used. Scores were compared over 4 years, and a Wilcoxon rank sum analysis was performed to determine significance. Evaluations were compared for 1999 and 2000, which were the 2 years before the initiation of the structured rotation, with 2001 and 2002, the first 2 years of the new approach's implementation. RESULTS: Sixty third-year students were evaluated. The students noted that the quality of teaching improved (P <.002), the instructors' commitment and enthusiasm increased (P <.001), instructors enhanced the student's professional development (P <.001), and students perceived faculty as positive role models (P <.001). It is noteworthy that between the years 1999 and 2000, the number of students interested in obstetrics and gynecology was 3 and 4, respectively, whereas in 2001 and 2002, the first 2 years after implementation of the new process, 6 and 7 students, respectively, were interested in this field. CONCLUSION: Students' satisfaction with their third-year clerkship improved with a structured program and increased faculty involvement. LEVEL OF EVIDENCE: II-2 PMID- 14754706 TI - Operative note dictation: should it be taught routinely in residency programs? AB - OBJECTIVE: To determine the extent of formal education regarding operative dictation in U.S. Obstetrics and Gynecology residency programs and to prospectively evaluate the effectiveness of formal teaching regarding operative dictation. METHODS: A 1-page questionnaire was mailed to all U.S. Obstetrics and Gynecology residency program directors (n = 270). The operative dictations of all Obstetrics and Gynecology residents at the University of Iowa before and after a 30-minute formal teaching session were evaluated using a scoring system developed by the authors of this study (scale 0-20). RESULTS: A 73% response rate (n = 198) was obtained from the surveys. The results from the survey demonstrated that only 23% of programs provide formal teaching regarding operative dictations; however, 83% of the residency program directors felt that it is an important skill to teach. All 16 obstetrics and gynecology residents at the University of Iowa attended a 30-minute teaching session on operative dictation. The mean scores for all residents improved from 9.06 to 18.56 after a formal teaching session (P <.001). The preteaching scores comparing the 4 classes of residents also varied significantly (ranging from a score of 5.5 for first-year residents to 11.25 for the fourth-year residents, P =.009). There were no differences between scores among the residents after they attended the formal teaching session (P =.11). CONCLUSION: Formal teaching of operative dictation is uncommon in U.S. residency programs but felt to be important by most residency program directors. A brief teaching session is effective and may be useful during residency training. LEVEL OF EVIDENCE: II-3 PMID- 14754707 TI - Anesthetic effect of intrauterine lidocaine plus naproxen sodium in endometrial biopsy. AB - OBJECTIVE: To compare the efficacy of intrauterine lidocaine with oral naproxen sodium on pain perception of the patients during endometrial biopsy using the Pipelle instrument and to investigate their effects when used in combination. METHODS: One-hundred twenty women were randomly assigned to receive either 5 mL of intrauterine 2% lidocaine or saline and either 550 mg of naproxen sodium or a similar-appearing placebo tablet. Subsequently, each woman completed a 10-cm visual analog scale for subjective pain experience and a physician scored visible signs of the women's distress during the procedure using a 3-point observer scale. RESULTS: There was no statistically significant difference between the 4 groups in age, vaginal parity, history of chronic pelvic pain, menopausal status, tenaculum use, previous endometrial biopsy, or difficulty in passing the cervical os. The mean pain scores of the women in the naproxen only (5.8 +/- 2.2) and lidocaine only (5.9 +/- 2.2) groups were not significantly different compared with placebo group (7.1 +/- 2.0). However, the mean pain score in the lidocaine plus naproxen group (4.6 +/- 1.8) compared with the placebo group showed significant reduction in pain (P <.05). Pain rated by the physician was significantly lower in the lidocaine plus naproxen group compared with other groups, and a significant correlation was noted between the visual analog pain score and the patients' distress recorded by the physician (r =.791, P <.001). One patient in the naproxen-only group had vasovagal syncope after the procedure. CONCLUSION: Intrauterine lidocaine instillation significantly decreases pain associated with Pipelle endometrial biopsy when used in combination with oral naproxen sodium. LEVEL OF EVIDENCE: I PMID- 14754708 TI - Prevalence and severity of urinary incontinence in kidney transplant recipients. AB - OBJECTIVE: To estimate the prevalence and severity of urinary incontinence and its impact on activities of daily living in kidney transplant recipients. METHODS: The continence status, severity, and impact of urinary incontinence on activities of daily living in female kidney transplant recipients was studied by using Sandvik's Incontinence Severity Index and the Incontinence Impact Questionnaire. Clinical and sociodemographic data from post-transplant incontinent and continent participants were compared by using chi(2), Student t, and Mann-Whitney U tests. RESULTS: One hundred of 123 female kidney transplant recipients completed the survey for a response rate of 81.3%. The prevalence of urinary incontinence in female kidney transplant recipients was 28%. The mean Sandvik's Incontinence Severity Index score was 5.1 (median 3.5; 95% confidence interval 3.6, 6.6). The mean Incontinence Impact Questionnaire score was 12.9 (median 4.8; 95% confidence interval 6.7, 18.9). Incontinent kidney transplant recipients recorded a greater daily fluid intake on their 3-day diaries than continent kidney transplant recipients (2,951.9 +/- 1,228.6 mL compared with 1,854.6 +/- 614.8 mL, P =.04). CONCLUSIONS: The prevalence rate of urinary incontinence from interviewed kidney transplant recipients was 28%, which had minimal impact on activities of daily living and may not be predicted by a history of incontinence before kidney failure. LEVEL OF EVIDENCE: II-3 PMID- 14754709 TI - Combination estrogen-progestin contraceptives and body weight: systematic review of randomized controlled trials. AB - OBJECTIVE: Many women and clinicians believe that combination estrogen-progestin contraceptive use can lead to weight gain. This concern can deter women from starting hormonal contraception or lead to premature quitting. This review evaluated the association between combination contraceptive use and change in body weight. DATA SOURCES: The computerized databases CENTRAL, MEDLINE, EMBASE, Popline, and LILACS (from their inception to 2002) were used to conduct this review. Known investigators and manufacturers were contacted for information about other trials not discovered in the database search. METHODS OF STUDY SELECTION: All English-language, randomized controlled trials measuring weight change that were at least 3 treatment cycles in duration and that compared a combination contraceptive to a placebo or to a combination contraceptive that differed in drug, dosage, regimen, or study length were eligible for inclusion. Of the 570 reports of randomized controlled trials of eligible interventions that were identified, 42 trials were included in the systematic review. TABULATION, INTEGRATION, AND RESULTS: Two reviewers independently abstracted data from the eligible trials using a standard form. Depending on the data available, the weighted mean difference using a fixed effect model with 95% confidence intervals was calculated for the mean change in weight between baseline and posttreatment measurements or the Peto odds ratio with 95% confidence intervals was calculated by using the proportion of women who gained or lost more than a specified amount of weight. The 3 placebo-controlled, randomized trials did not find evidence supporting a causal association between combination oral contraceptives or a combination skin patch and weight gain. Most comparisons from the 40 trials that compared 2 or more combination contraceptives showed no substantial difference in weight. In addition, discontinuation of combination contraceptives because of weight gain did not differ between groups when this factor was studied. CONCLUSION: Available evidence is insufficient to determine the effect of combination contraceptives on weight, but no large effect is evident. PMID- 14754710 TI - Minimizing the risk of neurologic injury in gynecologic surgery. AB - OBJECTIVE: The association of intraoperative neurologic injuries with gynecologic surgical procedures is well established. The sequelae of such injuries are usually transient and resolve with minimal intervention, although long-term disability can and does occasionally occur. The purpose of this study was to examine the mechanisms by which these injuries occur in order to reduce the risk of their occurrence. DATA SOURCES: A MEDLINE search was performed cross referencing the index terms "neurological injury" and "gynecological surgery," from January 1, 1960 to December 31, 2002. METHODS OF STUDY SELECTION: This article, based on the data and results (Level I-III) obtained from the MEDLINE search, examined the most common neurologic injuries that occur in association with abdominal and vaginal surgical procedures routinely performed by gynecologists. TABULATION, INTEGRATION, AND RESULTS: Neurologic injuries after pelvic surgery all generally share a common etiology, specifically injury to one or more components of the lumbosacral nerve plexus. Three major factors that predispose to neurologic injury at the time of gynecological surgery are 1) the improper placement or positioning of self-retaining or fixed retractors, particularly those with deep lateral retractor blades; 2) improper positioning of patients in lithotomy position preoperatively; and 3) radical surgical dissection resulting in autonomic nerve disruption. Level I data strongly implicate the improper placement of self-retaining or fixed retractors as the most common cause of femoral nerve injury arising in association with abdominal surgical procedures. CONCLUSION: A thorough understanding of the anatomy of the lumbosacral nerve plexus and the mechanisms by which operative injuries to this plexus occur will enable the gynecologic surgeon to reduce the subsequent risk of their occurrence in his or her own surgical practice. PMID- 14754711 TI - Laparoscopic exposure in obese high-risk patients with mechanical displacement of the abdominal wall. AB - BACKGROUND: Patients with morbid obesity or pulmonary disease are at a higher risk for complications during advanced laparoscopic procedures. Higher intraperitoneal carbon dioxide pressures required to elevate the pannus can negatively impact hemodynamic and respiratory parameters. CASES: We describe a technique that uses a combination of a mechanical retractor and a Foley catheter inserted midway between the umbilicus and the pubic symphysis that assists in elevating the anterior abdominal wall. In 3 cases this technique allowed for a low-pressure pneumoperitoneum during advanced laparoscopic pelvic surgery, which resulted in improved hemodynamic parameters and pulmonary function in these high risk patients. CONCLUSION: The Foley Lap-Lift facilitated laparoscopy through mechanical abdominal wall elevation and allowed for a lower-pressure pneumoperitoneum. This technique is an addition to traditional operative laparoscopy in select high-risk patients. PMID- 14754712 TI - Ethical dimensions of elective primary cesarean delivery. AB - Cesarean deliveries are among the most common surgical procedures performed in the United States. Recent publications demonstrate the reduced risks of these operations and describe their potential benefits to both mothers and children. Recent surveys show that a substantial minority of obstetricians would accede to patients' requests for elective primary cesarean delivery, and some of these professionals would prefer that mode of delivery for themselves or their partners. However, scant attention has been paid to the ethical underpinnings of surgery by choice in these circumstances or ethically justified criteria for determining the role of patient choice in elective surgery generally. We define and elaborate upon the role of beneficence-, autonomy-, and justice-based considerations in these deliberations. We conclude that beneficence-based clinical judgment still favors vaginal delivery. Additionally, we have no confidence that either offering or performing elective cesarean delivery is consistent with substantive-justice-based considerations and conclude that there is no autonomy-based obligation to offer cesarean delivery in an ethically and legally appropriate informed consent process. Physicians should respond to patient-initiated requests for such procedures with a thorough informed consent process and request that the woman reconsider to ensure that her autonomy is being meaningfully exercised. In such cases, implementing a woman's request is ethically permissible. PMID- 14754714 TI - Evaluating evaluation forms form. AB - OBJECTIVES: To provide a tool for evaluating evaluation forms. METHODS: A new form has been developed and tested on itself and a sample of evaluation forms obtained from the graduate medical education offices of several local universities. Additional forms from hospital administration were also subjected to analysis. RESULTS: The new form performed well when applied to itself. The form performed equally well when applied to the other (subject) forms, although their scores were embarrassingly poor. CONCLUSIONS: A new form for evaluating evaluation forms is needed, useful, and now available. PMID- 14754715 TI - Asymmetric distribution of sciatic nerve endometriosis. PMID- 14754716 TI - Is the intrapartum biophysical profile useful? PMID- 14754717 TI - Superiority of electrocautery over the suture method for achieving cervical cone bed hemostasis. PMID- 14754718 TI - ACOG Committee Opinion. Uterine artery embolization. AB - Uterine artery embolization for the treatment of symptomatic uterine leiomyomata has become increasingly popular. Based on current evidence, it appears that uterine artery embolization, when performed by experienced physicians, provides good short-term relief of bulk-related symptoms and a reduction in menstrual flow. Complication rates associated with the procedure are low, but in rare cases can include hysterectomy and death. There is insufficient evidence to ensure its safety in women desiring to retain their fertility, and pregnancy-related outcomes remain understudied. The American College of Obstetricians and Gynecologists' Committee on Gynecologic Practice considers the procedure investigational or relatively contra-indicated in women wishing to retain fertility. The use of uterine artery embolization in postmenopausal women is rarely, if ever, indicated. The Committee strongly recommends that women who wish to undergo uterine artery embolization have a thorough evaluation with an obstetrician-gynecologist to help facilitate optimal collaboration with interventional radiologists and to ensure the appropriateness of this therapy, taking into account the reproductive wishes of the patient. It is also recommended that all patients considering uterine artery embolization be adequately informed about potential complications. PMID- 14754719 TI - The match. PMID- 14754720 TI - Prospective evaluation of thermal capsulorrhaphy for shoulder instability: indications and results, two- to five-year follow-up. AB - BACKGROUND: Thermal shrinkage of capsular tissue has recently been proposed as a means to address the capsular redundancy associated with shoulder instability. Although this procedure has become very popular, minimal peer-reviewed literature is available to justify its widespread use. PURPOSE: To prospectively evaluate the efficacy of arthroscopic electrothermal capsulorrhaphy for the treatment of shoulder instability. STUDY DESIGN: This nonrandomized prospective study evaluated the indications and results of thermal capsulorrhaphy in 84 shoulders with an average follow-up of 38 months. METHODS: Patients were divided into three clinical subgroups: traumatic anterior dislocation (acute or recurrent), recurrent anterior anterior/inferior subluxation without prior dislocation, and multidirectional instability. Patients underwent arthroscopic thermal capsulorrhaphy after initial assessment, radiographs, and failure of a minimum of 3 months of nonoperative rehabilitation. RESULTS: Outcome measures included pain, recurrent instability, return to work/sports, and the American Shoulder and Elbow Surgeons (ASES) Shoulder Assessment score. Overall results were excellent in 33 participants (39%), satisfactory in 20 (24%), and unsatisfactory in 31 (37%). CONCLUSIONS: The high rate of unsatisfactory overall results (37%), documented with longer follow-up, is of great concern. The authors conclude that enthusiasm for thermal capsulorrhaphy should be tempered until further studies document its efficacy. PMID- 14754721 TI - Does the patellar tendon normalize after harvesting its central third?: a prospective long-term MRI study. AB - BACKGROUND: The central third of the patellar tendon is the most frequently used autograft for ACL reconstruction. HYPOTHESIS: The patellar tendon at the donor site would look normal as seen on MRI 6 years after harvesting its central third. METHODS: Nineteen consecutive patients were included in the study. MRI examinations of the donor site were performed at 6 (5 to 10) weeks, 6 (6 to 8) months, 27 (24 to 29) months, and 71 (68 to 73) months postoperatively. The contralateral normal side was examined only on the first occasion. RESULTS: The size of the donor-site gap decreased significantly (P = 0.0001) between 6 weeks and 6 years. In most patients, a thinning of the central part of the patellar tendon was still found at 6 years. The thickness of the peripheral patellar tendon was increased, compared with the contralateral healthy side, until 2 years (P = 0.003). On all occasions, the width was increased compared with the contralateral side (P < 0.015). CONCLUSION: Prospective MRI examinations revealed that the patellar tendon at the donor site had not normalized 6 years after harvesting its central third. The reharvesting of the patellar tendon can therefore not be recommended. PMID- 14754722 TI - Ultrasonographic and power Doppler evaluation of the patellar tendon ten years after harvesting its central third for reconstruction of the anterior cruciate ligament: comparison of patients without or with anterior knee pain. AB - BACKGROUND: The long-term morphologic changes in the patellar tendon after harvesting its central third for the reconstruction of the anterior cruciate ligament have not been reported. HYPOTHESIS: Sonographic morphologic changes in the harvested patellar tendon are common 10 years after the harvesting procedure. STUDY DESIGN: Retrospective cohort study. METHODS: Thirty-one patients who had undergone an anterior cruciate ligament reconstruction using central-third bone patellar tendon-bone autograft with a closure of the patellar tendon defect were included in this study. An ultrasonographic and Power Doppler examination was performed at a mean follow-up of 10 years. RESULTS: Ultrasonography of the harvested patellar tendon showed intratendinous calcification in 9 patients, hypoechoic lesion in 20 patients, hyperechoic lesion in 1 patient, and peritendinous changes in 1 patient. Only 3 (9.7%) of the 31 patients had no changes in the harvested patellar tendon. No abnormality was visible in the contralateral (normal) patellar tendons of the 31 patients. The harvested patellar tendon was significantly thicker than the contralateral patellar tendon (P < 0.05). CONCLUSIONS: Sonographic morphologic changes of the patellar tendon were common 10 years after the harvesting procedure, although no evidence of inflammatory process, as could be seen in Power Doppler flow, was found inside of any patellar tendon. PMID- 14754723 TI - Grade 1 or "ding" concussions in high school athletes. AB - BACKGROUND: Recent concussion management guidelines have suggested that athletes with mild (grade 1) concussions may be returned to play if asymptomatic for 15 minutes. The purpose of this study was to assess the utility of a current concussion management guideline in classifying and managing mild concussion. HYPOTHESIS: High school athletes diagnosed with a grade 1 concussion will demonstrate measurable decline in neuropsychological functioning that persists during the 1st week of recovery. STUDY DESIGN: Prospective study designed to evaluate neuropsychological functioning both prior to and following concussion. METHODS: Forty-three high school athletes completed neuropsychological test performance and symptom ratings prior to the season and at two times during the 1st week following mild concussion. RESULTS: Thirty-six hours after injury, mildly concussed high school athletes demonstrated a decline in memory (P < 0.003) and a dramatic increase in self-reported symptoms (P < 0.00001) compared to baseline performance. CONCLUSIONS: Athletes with grade 1 concussion demonstrated memory deficits and symptoms that persisted beyond the context in which they were injured. These data suggest that current grade 1 return-to-play recommendations that allow for immediate return to play may be too liberal. CLINICAL RELEVANCE: A reconsideration of current concussion grading systems appears to be warranted. PMID- 14754724 TI - MR imaging of the hip and knee before and after marathon running. AB - BACKGROUND: Despite its obvious benefits regarding aerobic fitness, the possible deleterious effects of long-distance running remain controversial. The repetitive loading associated with this activity could potentially predispose to the subsequent development of osteoarthritis. Lower extremity malalignment can also result in abnormal joint loading and is another possible contributing factor for premature articular cartilage degeneration. The purpose of this study is to investigate whether external impact loading in marathon runners creates internal stresses on bone and cartilage that are demonstrable on MR images. Participants were separately assessed for static lower extremity alignment, using standard radiographs. METHODS: Six recreational and two semi-professional runners underwent magnetic resonance (MR) imaging of the hip and knee before and after a marathon run using coronal T1-weighted and coronal STIR sequences. RESULTS: The pre-run and post-run scans failed to demonstrate marrow oedema, periosteal stress reactions, or joint effusions in seven runners. One patient who underwent a reconstruction of his anterior cruciate ligament 18 months ago demonstrated a small effusion in the reconstructed knee before and after the race. CONCLUSIONS: Our results suggest that the high impact forces in long-distance running are well tolerated and subsequently do not demonstrate changes on MR images. PMID- 14754725 TI - Opening wedge high tibial osteotomy for symptomatic hyperextension-varus thrust. AB - BACKGROUND: The purpose of this study was to assess the functional outcome of opening wedge high tibial osteotomy (HTO) in a young, active group of patients with instability rather than osteoarthritis. METHODS: The results of 17 opening wedge HTOs in 16 patients with a symptomatic hyperextension-varus thrust were evaluated. Functional results were evaluated according to the activity scoring system of Tegner and Lysholm and using a 5-point visual analogue scale to assess change in knee stability and satisfaction. Radiographs were analyzed to determine changes in femorotibial and mechanical axis alignment, tibial slope, and patellar height. RESULTS: Patients were followed for a mean of 56 months. All patients had an increase in their activity score postoperatively. Nine patients rated their symptoms as significantly better and seven as somewhat better. All but one were satisfied with the surgery. Femorotibial axis alignment was changed to a mean of 6 degrees valgus, mechanical axis alignment was corrected to a mean of 46% toward the lateral compartment, posterior tibial slope was increased a mean of 8 degrees, and the ratio of patellar height was decreased a mean of 0.17. CONCLUSION: Opening wedge HTO can produce good functional and radiographic results in selected patients with a symptomatic thrust. PMID- 14754726 TI - Biomechanical properties of patellar and hamstring graft tibial fixation techniques in anterior cruciate ligament reconstruction: experimental study with roentgen stereometric analysis. AB - BACKGROUND: Reliable fixation of the soft hamstring grafts in ACL reconstruction has been reported as problematic. HYPOTHESIS: The biomechanical properties of patellar tendon (PT) grafts fixed with biodegradable screws (PTBS) are superior compared to quadrupled hamstring grafts fixed with BioScrew (HBS) or Suture-Disc fixation (HSD). STUDY DESIGN: Controlled laboratory study with roentgen stereometric analysis (RSA). METHODS: Ten porcine specimens were prepared for each group. In the PT group, the bone plugs were fixed with a 7 x 25 mm BioScrew. In the hamstring group, four-stranded tendon grafts were anchored within a tibial tunnel of 8 mm diameter either with a 7 x 25 mm BioScrew or eight polyester sutures knotted over a Suture-Disc. The grafts were loaded stepwise, and micromotion of the graft inside the tibial tunnel was measured with RSA. RESULTS: Hamstring grafts failed at lower loads (HBS: 536 N, HSD 445 N) than the PTBS grafts (658 N). Stiffness in the PTBS group was much greater compared to the hamstring groups (3500 N/mm versus HBS = 517 N/mm and HSD = 111 N/mm). Irreversible graft motion after graft loading with 200 N was measured at 0.03 mm (PTBS), 0.38mm (HBS), and 1.85mm (HSD). Elasticity for the HSD fixation was measured at 0.67 mm at 100 N and 1.32 mm at 200 N load. CONCLUSION: Hamstring graft fixation with BioScrew and Suture-Disc displayed less stiffness and early graft motion compared to PTBS fixation. Screw fixation of tendon grafts is superior to Suture-Disc fixation with linkage material since it offers greater stiffness and less graft motion inside the tibial tunnel. CLINICAL RELEVANCE: Our results revealed graft motion for hamstring fixation with screw or linkage material at loads that occur during rehabilitation. This, in turn, may lead to graft laxity. PMID- 14754727 TI - A clinical and roentgenographic study of Little League elbow. AB - BACKGROUND: Comparisons of medial elbow injury rate and the incidence of clinical and radiographic findings among Little League baseball players have not been documented. HYPOTHESIS: Injury rate and clinical and radiographic findings in Little Leaguers of different positions may be similar. STUDY DESIGN: Survey and retrospective review. METHODS: Altogether, 343 Little Leaguers (120 pitchers, 40 catchers, and 183 fielders) participated in the study. The survey (including age, playing position, playing experience, and symptoms), clinical examination, and roentgenogram of both elbows were obtained for analysis using Pearson's chi square test. RESULTS: Clinically, 58% of the pitchers presented with soreness compared to 63% in catchers and 47% in fielders. Radiographic examination revealed hypertrophy of the medial humeral epicondyle in all pitchers and catchers and 90% of the fielders. Separation of the medial epicondyle was found in 63% of pitchers, 70% of catchers, and 50% of fielders, while fragmentation was found in 19% of pitchers, 40% of catchers, and 15% of fielders. In subjects with separation or fragmentation, 49% and 56% complained of elbow soreness, respectively. CONCLUSION: Similar rates of medial elbow changes in Little Leaguers may imply that the roentgenological changes of the medial epicondyle could be an adaptive yet physiological reaction to the excessive valgus stress of throwing. The higher incidence of medial epicondylar separation and fragmentation in catchers that has not been reported before may be secondary to the increased stress placed on the elbow when throwing from the squatting position. PMID- 14754728 TI - The medial collateral ligament of the elbow is not isometric: an in vitro biomechanical study. AB - BACKGROUND: The anterior bundle of the medial collateral ligament (AMCL) of the elbow has been shown to be the most important valgus stabilizer of the elbow. However, the isometry of this band has not been quantified. HYPOTHESIS: Isometric fibers exist within the AMCL, and these fibers are located within its central region. STUDY DESIGN: Controlled laboratory study. METHODS: Twelve cadaveric elbow specimens were mounted in a testing apparatus in a valgus gravity-loaded orientation. Passive supinated flexion was performed and the motion recorded using an electromagnetic tracking device. Hundreds of attachment points for the AMCL of the elbow were recorded on the medial epicondyle and ulna. The overall change in length between each point on the ulna to every humeral point, throughout the arc of motion, was quantified (DeltaL = Lmax - Lmin). The locations of the smallest DeltaL values were determined relative to the attachment site of the AMCL on the medial epicondyle. RESULTS: True isometry was not found throughout the arc of flexion. The smallest DeltaL values averaged 2.8 +/- 1.2 mm (range: 0.7 mm to 5.2 mm). Isometric fibers do not exist within the AMCL; however, "nearly" isometric areas are located on the lateral aspect of the attachment site of the AMCL on the medial epicondyle, near the anatomic axis of rotation. CONCLUSIONS: We postulate that these nearly isometric areas would be the most ideal location for graft attachment during reconstruction of the AMCL. PMID- 14754729 TI - Patellar tendon-to-bone healing using high-density collagen bone anchor at 4 years in a sheep model. AB - BACKGROUND: This study reports the long-term histologic and mechanical properties of the healing patellar tendon-bone interface reconstructed using a high-density type I collagen bone anchor (HDC) compared to a metal anchor in sheep. HYPOTHESIS: To determine the long-term histology and mechanical properties of extra-articular tendon-bone healing and in vivo response to a HDC anchor. STUDY DESIGN: Controlled laboratory study. METHODS: The structural properties, tendon bone histology, and device histology in the bone were examined out to 208 weeks in a sheep model. RESULTS: The patellar tendon-proximal tibia bone interface continued to remodel with time but, by 4 years, had yet to develop the well defined zones of tendon, fibrocartilage, calcified cartilage, and bone of the native patellar tendon to bone insertion. The insertion repair strength did not vary between the repaired tendons and the nonoperated controls at any time. CONCLUSION: The healing tendon-bone interface undergoes a gradual remodeling process, which had yet to reconstitute back to the control interface by 208 weeks. The HDC device remained essentially intact at 208 weeks showing little signs of degradation. CLINICAL RELEVANCE: Tendon-bone healing is mechanically equivalent to the contralateral side by 26 weeks whereas histologic structure requires much longer to remodel back to the native insertion. PMID- 14754730 TI - Increased injury risk among first-day skiers, snowboarders, and skiboarders. AB - BACKGROUND: Inexperience predisposes skiers, snowboarders, and skiboarders to a higher risk of injury. HYPOTHESIS: Individuals on their very 1st day's experience of skiing, snowboarding, or skiboarding are at greater risk of injury. STUDY DESIGN: Case control study. METHODS: All individuals injured at the three largest ski areas in Scotland during the three winter seasons 1999-2000 through 2001-2002 were included. First-day participants were identified. Individual demographic details and snow sports-related parameters were recorded. Control data were collected from uninjured individuals. Factors associated with injury were explored in univariate and multivariate analyses. RESULTS: First-day participants were overrepresented in the injured population by a factor of 2.2. Participant age younger than 17 years or older than 26 years, snowboarding, taking professional instruction, and the use of rented or borrowed gear were all independently associated with increased injury risk among 1st-day participants. CONCLUSIONS: First-day participants are at increased risk of injury. Several independent risk factors for injury have been identified. CLINICAL RELEVANCE: First-day participants should be targeted in any future educational programs with emphasis on gear selection, use of protective gear, and the risks of exceeding the limits of their ability. PMID- 14754731 TI - Hip adductor injury in competitive swimmers. AB - METHODS: The authors conducted a survey of 296 competitive swimmers to assess the incidence and importance of interfering groin pain in breaststroke swimmers. RESULTS: Breaststroke swimmers were more likely to have current groin pain (6.92%) than individual medley swimmers who did not compete in pure breaststroke events (0, P = 0.015). Breaststroke swimmers (42.7%) were also more likely than individual medley swimmers (21.5%, P = 0.000622) or nonbreaststroke, nonindividual medley swimmers (5.8%, P = 0.00000311) to have been unable to train breaststroke in practice during the past year due to groin injury. CONCLUSION: Results indicate that breaststroke swimmers are at significant risk of groin injury, groin injury is positively correlated with increased magnitude of breaststroke training, and groin injury may prevent participation in practices and competitions. PMID- 14754732 TI - The effect of posterior cruciate ligament reconstruction on patellofemoral contact pressures in the knee joint under simulated muscle loads. AB - BACKGROUND: The mechanism of cartilage degeneration in the patellofemoral joint (PFJ) and medial compartment of the knee following posterior cruciate ligament (PCL) injury remains unclear. PCL reconstruction has been recommended to restore kinematics and prevent long-term degeneration. The effect of current reconstruction techniques on PFJ contact pressures is unknown. PURPOSE: To measure PFJ contact pressures after PCL deficiency and reconstruction. METHOD: Eight cadaveric knees were tested with the PCL intact, deficient, and reconstructed. Contact pressures were measured at 30 degrees, 60 degrees, 90 degrees, and 120 degrees of flexion under simulated muscle loads. Knee kinematics were measured by a robotic testing system, and the PFJ contact pressures were measured using a thin film transducer. A single bundle achilles tendon allograft was used in the reconstruction. RESULTS: PCL deficiency significantly increased the peak contact pressures measured in the PFJ relative to the intact knee under both an isolated quadriceps load of 400 N and a combined quadriceps/hamstrings load of 400 N/200 N. Reconstruction did not significantly reduce the increased contact pressures observed in the PCL-deficient knee. CONCLUSION: The elevated contact pressures observed in the PCL-deficient knee and reconstructed knee might contribute to the long-term degeneration observed in both the non-operatively treated and PCL-reconstructed knees. PMID- 14754733 TI - Infraspinatus muscle atrophy in professional baseball players. AB - BACKGROUND: Infraspinatus muscle atrophy has been observed in athletes who stress their upper extremities in an overhead fashion. The majority of such case reports have been in volleyball players, with far fewer cases reported in baseball players. HYPOTHESIS: Infraspinatus muscle atrophy occurs to a notable degree in professional baseball players. STUDY DESIGN: Retrospective cohort study. METHODS: At the end of the 1999 baseball season, data were collected from all Major League Baseball teams in regards to players affected with infraspinatus muscle atrophy. RESULTS: Twelve of the 1491 major league professional baseball players were identified as having appreciable infraspinatus muscle atrophy. There was an increased prevalence of the muscle atrophy in professional pitchers (10 of 494, 4%) compared to position players (2 of 997, 0.2%) (P <0.001). Among affected pitchers, the atrophy was identified more frequently in starting pitchers (8 of 10) compared to relief pitchers (2 of 10) (P = 0.036), pitchers who had played for more years at the major league level (8.7 +/- 4.9 versus 5.2 +/- 4.0) (P = 0.017), and pitchers who had thrown for more innings at the major league level (971.4 +/- 784.4 versus 485.0 +/- 594.6) (P <0.001). CONCLUSION: Infraspinatus atrophy was identified in 4.4% of major league starting pitchers and occurred in those pitchers who pitched for more years and innings during their major league career. PMID- 14754734 TI - Prevalence of the Bennett lesion of the shoulder in major league pitchers. AB - BACKGROUND: The Bennett lesion is a mineralization of the posterior inferior glenoid noted in overhead throwing athletes. Although previous studies have debated appropriate treatment of the lesion, no studies have indicated the lesion prevalence in throwing athletes. HYPOTHESIS: The Bennett lesion is more common than previously believed and may represent an asymptomatic finding. STUDY DESIGN: Uncontrolled retrospective review. METHODS: Fifty-five asymptomatic major league pitchers underwent routine preseason radiographic screening. Radiographs were reviewed for the presence of a Bennett's lesion. Player demographics, pitching, and baseball records were reviewed to obtain the patient's dominant arm, age, years and innings pitched, and time on the disabled list or surgery. RESULTS: Twelve pitchers (22%) were noted to have a radiographic Bennett lesion. No statistically significant difference was noted in age, years pitched, or innings pitched between pitchers with and without a Bennett lesion. No player who demonstrated a Bennett lesion required surgical treatment for shoulder pain during his time with the club. Two players required time on the disabled list, but neither player had complaints of posterior shoulder pain. CONCLUSIONS: This lesion is a relatively common finding in major league pitchers. Concomitant pathology should be suspected when evaluating throwers with posterior shoulder pain and this lesion. PMID- 14754735 TI - Chondrocyte viability in refrigerated osteochondral allografts used for transplantation within the knee. AB - PURPOSE: To evaluate cell viability and matrix characteristics of refrigerated osteochondral allografts implanted up to 44 days after harvest. METHODS: Sixteen refrigerated allografts underwent histologic and ultrastructural examination and fluorescence excitation analysis prior to implantation. The average size of the graft implanted was 6.2 cm(2) (+/-3.4 cm(2)). Refrigerated allografts averaged 30 days (range, 17 to 44 days) from donor expiration to implantation. Nine specimens underwent cell viability testing. The percent viability of refrigerated allografts prior to implantation averaged 67%. RESULTS: No significant correlations were noted between histologic score, electron microscopy score, matrix staining percent (MSP) score, and viability. When time to implantation was assessed, an inverse correlation was noted with MSP score (r =.539) (P < 0.05), indicating less matrix staining in grafts refrigerated longer after harvest. CONCLUSION: The current data indicate that refrigerated osteochondral allografts can be maintained for up to 44 days with average chondrocyte viability of 67%. PMID- 14754736 TI - Chondrocyte survival and material properties of hypothermically stored cartilage: an evaluation of tissue used for osteochondral allograft transplantation. AB - BACKGROUND: There is little information available on the material properties of hypothermically stored allograft specimens used to repair osteochondral defects. PURPOSE: To analyze the effect of hypothermic storage on the material properties of fresh knee specimens over a 60-day interval. STUDY DESIGN: Controlled laboratory study. METHODS: Twelve sheep knee condyles were isolated. The femoral and tibial condyles and the patella were stored in a nutritive medium containing Dulbecco's Modified Eagle's Medium for 1, 8, 15, 29, 45, or 60 days. Total chondrocyte density, chondrocyte viability, matrix proteoglycan content, matrix water content, and matrix dynamic modulus of elasticity were determined. RESULTS: Mean chondrocyte viability decreased significantly over the storage interval: 100% (day 1), 98.2% (day 8), *80.2% (day 15), *80.6% (day 29), *64.6% (day 45), and *51.6% (day 60) (* P < 0.05). Qualitative analysis demonstrated a preponderance of nonviable chondrocytes in the superficial cartilage layer. Significant decreases in matrix proteoglycan were observed in day 15 through day 60 specimens (P < 0.05). The matrix dynamic modulus significantly decreased from day 1 through day 60 (P < 0.05). CONCLUSION: The material properties of hypothermically stored knee condyles progressively decline over 60 days. CLINICAL RELEVANCE: This observed decline may have significant ramifications on long-term graft survival following stored-allograft implantation. PMID- 14754737 TI - Characteristic plantar pressure distribution patterns during soccer-specific movements. AB - PURPOSE: To characterize in-shoe pressure measurements during different soccer specific maneuvers on two playing surfaces to identify the main loading areas of the foot. METHODS: Twenty-one experienced male soccer players participated in the study (25.5 +/- 1.8 years, 78.7 +/- 5.4 kg, and 182.9 +/- 5.7 cm). The Pedar Mobile system was used to collect plantar pressure information inside the soccer shoe. Four soccer-specific movements were performed (normal run, cutting maneuver, sprint, and goal shot) on both a grass and a red cinder surface. RESULTS: Results showed characteristic pressure distribution patterns with specific loading areas of the foot that correspond to the evaluated movements. In addition, loading patterns with higher pressure values than those observed during normal run were found. In cutting, the medial part of the foot; in sprinting, the first and second ray; and in kicking, the lateral part of the foot are predominantly loaded. No global effect of the two surfaces on pressure parameters was found. CONCLUSION: The results of the present investigation suggest that the high load in soccer in combination with a high repetition may have an important influence in the development of overuse injuries. PMID- 14754738 TI - Cell-based therapy for meniscal repair: a large animal study. AB - BACKGROUND: The avascular portion of the meniscus cartilage in the knee does not have the ability to repair spontaneously. HYPOTHESIS: Cell-based therapy is able to repair a lesion in the swine meniscus. STUDY DESIGN: Controlled laboratory study. METHODS: Sixteen Yorkshire pigs were divided into four groups. A longitudinal tear was produced in the avascular portion of the left medial meniscus of 4 pigs. Autologous chondrocytes were seeded onto devitalized allogenic meniscal slices and were secured inside the lesion with two sutures. Identical incisions were created in 12 other pigs, which were used as three separate control groups: 4 animals treated with an unseeded scaffold, 4 were simply sutured, and 4 were left untreated. Meniscal samples were collected after 9 weeks, and the samples were analyzed grossly, histologically, and histomorphometrically. RESULTS: Gross results showed bonding of the lesion margins in the specimens of the experimental group, whereas no repair was noted in any of the control group specimens. Histological and histomorphometrical analysis showed multiple areas of healing in the specimens of the experimental group. CONCLUSIONS: This study demonstrated the ability of seeded chondrocytes to heal a meniscal tear. CLINICAL RELEVANCE: Cell-based therapy could be a potential tool for avascular meniscus repair. PMID- 14754739 TI - Augmentation of femoral fixation in hamstring anterior cruciate ligament reconstruction with a bioabsorbable bead: a prospective single-blind randomized clinical trial. AB - BACKGROUND: The EndoPearl is an adjunct to bioabsorbable interference screw fixation in the femoral tunnel in anterior cruciate ligament (ACL) reconstruction. The purpose of the study was to assess the clinical effectiveness of the EndoPearl using the KT-1000 Knee Arthrometer and the Mohtadi ACL Quality of Life (ACL-QOL) Questionnaire. HYPOTHESIS: The application of the EndoPearl in hamstring ACL reconstruction has no significant benefit when compared to conventional treatment in KT-1000 and ACL-QOL Questionnaire outcomes. STUDY DESIGN: Prospective single-blind randomized clinical trial. METHODS: ACL reconstruction with autogenous semitendinosus and gracilis tendons was performed on 35 patients. Preoperatively, 3 months, 6 months, and 18 months postoperatively, patients' knees were evaluated using the KT-1000 Knee Arthrometer and the ACL-QOL Questionnaire. RESULTS: Statistically significant differences were achieved in terms of KT-1000 side-to-side differences between the study group and the control group at 6 months and 18 months postoperatively; the mean side-to-side differences are significantly less in the study group when compared to the control group. Statistically significant differences were not detected when comparing the ACL-QOL Questionnaire between the two groups at all time intervals. CONCLUSION: The application of the EndoPearl in conjunction with a bioscrew in the femoral tunnel in autogenous ACL reconstruction using semitendinosus and gracilis tendon grafts provides a significantly decreased laxity up to 18 months postoperatively in terms of KT-1000 side-to-side differences. PMID- 14754740 TI - Wakeboarding injuries. AB - BACKGROUND: Wakeboarding is a popular water sport that has the potential to produce serious injuries. To date, there has been only one article describing an injury caused by wakeboarding. HYPOTHESIS: Wakeboarding injuries are common. STUDY DESIGN: Analysis of data obtained from physician and patient questionnaires. METHODS: Analysis of questionnaire data obtained from 156 orthopaedic surgeons and 86 wakeboarders. RESULTS: Of 156 orthopaedic surgeons completing a wakeboarding injuries questionnaire, 49% reported no wakeboarding injuries, 36% (57) reported treating at least 1 wakeboarding injury, and 15% had never heard of wakeboarding. Fifty-seven orthopaedic surgeons reported 122 injuries, of which 47% were either anterior cruciate ligament (ACL) tears (31%) or shoulder dislocations (15%). In addition, 21% of all the injuries were some type of fracture. Of 86 wakeboarders completing the injuries survey, 77% reported sustaining an injury. There were 82 reported injuries in this group, of which 34% were either ACL tears (17%) or ankle sprains (17%). CONCLUSIONS: This survey of wakeboarding injuries demonstrates that serious injuries can occur from participation in this sport. The most common injuries reported were ACL tears, shoulder dislocations, and ankle sprains. In addition, 21% of all the injuries were some type of fracture. PMID- 14754741 TI - Calcific insertional Achilles tendinopathy: reattachment with bone anchors. AB - BACKGROUND: Recalcitrant calcific insertional Achilles tendinopathy is difficult to treat. HYPOTHESIS: Bursectomy, excision of the distal paratenon, disinsertion of the tendon, removal of the calcific deposit, and reinsertion of the Achilles tendon with bone anchors is safe and effective. STUDY DESIGN: Longitudinal study. METHODS: Twenty-one patients (six women) (21 feet) (average age 46.9 +/- 6.4 years) with recalcitrant calcific insertional Achilles tendinopathy were treated surgically with removal of the calcific deposit; the Achilles tendon was reinserted with bone anchors. RESULTS: At an average follow-up of 48.4 months, one patient necessitated a further operation. Eleven patients reported an excellent result, and five a good result. The remaining five patients could not return to their normal levels of sporting activity and kept fit by alternative means. The results of the VISA-A questionnaire were markedly improved in all patients, from an average of 62.4% to 88.1%. CONCLUSIONS: We recommend disinsertion of the Achilles tendon to excise the calcific deposit fully and reinsertion of the Achilles tendon in the calcaneus with suture anchors. No patient experienced a traumatic disinsertion of the reattached tendon. However, five patients were not able to return to their original level of physical activity. PMID- 14754742 TI - Medial ankle instability: an exploratory, prospective study of fifty-two cases. AB - BACKGROUND: Very little objective data are available regarding medial ankle instability. HYPOTHESIS: Several structures contribute to the stabilization of the medial ankle, and, in the case of injury, they are not involved in a uniform way. STUDY DESIGN: Explorative, prospective case series. METHODS: Fifty-one patients (52 ankles; males 27, females 25; age 36.4 [16 to 60] years) were surgically treated because of medial ankle instability. All clinical findings and structural changes, as found by arthroscopy and surgical exploration, were compared with the clinical diagnosis and then addressed for surgical reconstruction. RESULTS: Pain in the medial gutter was noted in all ankles (100%). Arthroscopy verified a clinically expected additional lateral instability in 40 ankles (77%). At 4.43 years (2 to 6.5 years) after surgical reconstruction, the clinical result was considered to be good/excellent in 46 cases (90%), fair in 4 cases (8%), and poor in 1 case (2%). CONCLUSION: The clinical characteristics of medial ankle instability are a feeling of giving way, pain on the medial gutter of the ankle, and a valgus and pronation deformity of the foot that can typically be actively corrected by the posterior tibial muscle. Arthroscopy was shown to be a very helpful diagnostic tool in verifying medial instability. PMID- 14754743 TI - Recovery of articular cartilage metabolism following thermal stress is facilitated by IGF-1 and JNK inhibitor. AB - BACKGROUND: The safety of intra-articular use of thermal probes is related to whether chondrocytes can tolerate exposure to high temperatures and whether cytoprotective agents may improve chondrocyte survival after thermal injury. PURPOSE: This study was conducted to characterize the metabolic responses of articular cartilage after short-term exposure to temperatures between 50 degrees C and 60 degrees C with and without addition of insulin-like growth factor 1 (IGF 1) and c-Jun N-terminal kinase (JNK) inhibitor. METHODS: Human articular cartilage from osteoarthritic knees was subjected to defined thermal stress. RESULTS: Although significant reduction of proteoglycan synthesis was observed after 5 seconds of exposure to 55 degrees C and 60 degrees C and after 10- to 30 second exposures to 53 degrees C, recovery of metabolic activity levels was observed after 7 days. CONCLUSION: Addition of IGF-1 and JNK inhibitor Sp600125 enabled the cartilage to maintain significantly higher levels of proteoglycan synthesis immediately after thermal stress. IGF-1 also enhanced recovery of metabolic activity after 7 days. CLINICAL SIGNIFICANCE: Results from this study indicate that there may be time and temperature parameters within which thermal chondroplasty can be safely performed. The data additionally suggest that inadvertent chondrocyte injury may be minimized through potential addition of substances like IGF-1 or JNK inhibitor. PMID- 14754744 TI - Bilateral anterior cruciate ligament reconstruction as a single procedure: evaluation of cost and early functional results. AB - BACKGROUND: The ideal treatment for patients presenting with bilateral anterior cruciate ligament (ACL) deficiency remains controversial. PURPOSE: To evaluate cost and early functional results after bilateral ACL reconstruction at a single setting. STUDY DESIGN: Retrospective review. METHODS: Eleven patients (22 knees) who underwent bilateral ACL reconstruction at a single setting were compared with 33 patients (35 knees) who underwent unilateral ACL reconstruction during the same time period. RESULTS: The mean time to full unrestricted activity between groups was 6.5 months for both groups (P = 0.92). There were no significant differences between groups at latest follow-up for complication rates or laxity as judged by Lachman test, pivot shift test, and KT 1000 arthrometry. The mean International Knee Documentation Committee subjective score at a mean 3.1-year follow-up was 91.9 for the bilateral ACL group compared to 92.0 for the unilateral ACL group (P = 0.95). There was a total cost savings per knee (based on 2001 dollars) of $3751.59 when performing bilateral ACL reconstruction at a single setting (P = 0.0001). CONCLUSIONS: For patients presenting with bilateral ACL deficient knees, reconstruction of both knees at a single setting is safe, cost effective, and does not appear to compromise early functional results. PMID- 14754745 TI - Delayed anterior cruciate ligament reconstruction in adolescents with open physes. AB - BACKGROUND: Treatment of anterior cruciate ligament rupture in adolescents with open physes is controversial. HYPOTHESIS: Delaying reconstruction until the physes bridge will affect the rate of additional knee injuries. STUDY DESIGN: Case control. METHODS: The physes group was 13 adolescents with open physes whose anterior cruciate ligament reconstructions were delayed until their physes bridged. Specific types of activity were absolutely restricted during the delay. The physes groups' rates of additional injuries, identified arthroscopically at reconstruction, were compared to rates among 116 skeletally mature adolescents. The comparison adolescents were stratified into four groups by interval from injury to reconstruction (<1 week, 1 to 6 weeks, 6 to 26 weeks, >26 weeks). RESULTS: The additional injury rates in the physes and four comparison groups were 46%, 50%, 47%, 43%, and 69%, respectively. Severity of additional injury, mechanism of injury, and rate of additional surgical procedures were similar among the groups. CONCLUSIONS: There was no evidence that intentionally delayed anterior cruciate ligament reconstruction increased the rate of additional knee injuries. Delayed reconstruction is a valid treatment option for adolescents with open physes at injury. Absolute activity restriction is key to decreasing the risk of additional knee injuries. PMID- 14754746 TI - Articular cartilage lesions in 993 consecutive knee arthroscopies. AB - BACKGROUND: Traumatic articular cartilage injuries heal poorly and may lead to development of osteoarthritis at a young age. This study estimates the number of patients who may benefit from one of the surgical methods of cartilage repair. METHODS: All patients undergoing knee arthroscopy during a 6-month period at three collaborating hospitals were consecutively evaluated according to the International Cartilage Repair Society (ICRS) knee form. The material consists of 993 consecutive knee arthroscopies in patients with median age of 35 years. RESULTS: Preoperative radiographs demonstrated degenerative changes in 13% of the knees. Articular cartilage pathology was found in 66% and a localized cartilage defect was found in 20% of the knees. A localized full-thickness cartilage lesion (ICRS grade 3 and 4) was observed in 11% of the knees. Of the localized full thickness lesions, 55% (6% of all knees) had a size above 2 cm(2). CONCLUSION: Eleven percent of all knee arthroscopies show cartilage defects that may be suitable for cartilage repair procedures. However, the natural history of these lesions and the number of patients that will benefit from a cartilage repair procedure are so far unknown. PMID- 14754747 TI - Sensorimotor control is impaired in dancers with functional ankle instability. AB - BACKGROUND: Factors potentially causing chronic instability after ankle inversion sprains have rarely been examined during the injuring movement. PURPOSE: To compare control of ankle movement during quiet stance and after inversion perturbation in chronically unstable ankles (n = 16) with healthy controls (n = 26). METHODS: Movement control was measured as magnitude of lateral ankle oscillation, using 3SPACE Fastrak during single leg stance (baseline oscillation) in two foot positions, flat and demi-pointe. In both positions, time to resume baseline oscillation after inversion perturbation (perturbation time) of 15 degrees for the flat foot and 7.5 degrees on demi-pointe was also determined. RESULTS: Baseline oscillation on demi-pointe was significantly smaller (P < 0.005) for the sprained group (2.5 +/- 0.5 mm) than for controls (4.0 +/- 2.3 mm). Perturbation time for the flat foot was significantly longer (P < 0.05) for the sprained group (2.2 +/- 0.4 seconds) than for controls (1.8 +/- 0.5 seconds). However, failure rate was higher (P < 0.05) among the sprained group than controls for perturbation with the foot flat and baseline oscillation on demi pointe. CONCLUSIONS: Findings demonstrated altered sensorimotor control in chronically unstable ankles. Those sprainers who successfully completed the tasks minimized oscillation. The impairments in the sprained group may reflect deficits in either movement detection, peroneal muscle response, or both. PMID- 14754748 TI - The effect of bracing on patellofemoral joint stress during free and fast walking. AB - BACKGROUND: Although several studies have demonstrated decreases in patellofemoral pain (PFP) with the application of bracing, the mechanism by which bracing reduces symptoms has not been elucidated. HYPOTHESIS: Individuals who responded favorably to bracing will exhibit decreased patellofemoral stress during level walking. STUDY DESIGN: Repeated measures, cross-sectional. METHODS: Fifteen subjects with a diagnosis of PFP completed two phases of data collection: 1) MRI assessment of patellofemoral contact area and 2) gait analysis. Data were obtained under braced and nonbraced conditions. Variables obtained from both data collection sessions were used as input variables into a mathematical model to quantify patellofemoral stress. RESULTS: Subjects reported a 56% reduction in pain following bracing. Bracing significantly reduced peak stress during free and fast walking (17% and 27%, respectively). The decrease in stress was the result of increased contact area as patellofemoral joint reaction forces were increased following bracing. CONCLUSION: Bracing resulted in a larger increase in patellofemoral contact area than the increase in joint reaction force, resulting in a decrease in joint stress. CLINICAL RELEVANCE: The results of this study suggest a possible mechanism by which bracing may be effective in reducing PFP and provides experimental support for the use of this treatment method. PMID- 14754750 TI - Bilateral fractures of the scapula in a professional football player: a case report. PMID- 14754749 TI - Late foreign-body reaction to Ticron sutures following inferior capsular shift: a case report. PMID- 14754751 TI - Adductor longus rupture in professional football players: acute repair with suture anchors: a report of two cases. PMID- 14754752 TI - An avulsion of the subscapularis in a skeletally immature patient. PMID- 14754753 TI - The use of ice in the treatment of acute soft-tissue injury: a systematic review of randomized controlled trials. AB - BACKGROUND: There are wide variations in the clinical use of cryotherapy, and guidelines continue to be made on an empirical basis. STUDY DESIGN: Systematic review assessing the evidence base for cryotherapy in the treatment of acute soft tissue injuries. METHODS: A computerized literature search, citation tracking, and hand searching were carried out up to April 2002. Eligible studies were randomized-controlled trials describing human subjects recovering from acute soft tissue injuries and employing a cryotherapy treatment in isolation or in combination with other therapies. Two reviewers independently assessed the validity of included trials using the Physiotherapy Evidence Database (PEDro) scale. RESULTS: Twenty-two trials met the inclusion criteria. There was a mean PEDro score of 3.4 out of of 10. There was marginal evidence that ice plus exercise is most effective, after ankle sprain and postsurgery. There was little evidence to suggest that the addition of ice to compression had any significant effect, but this was restricted to treatment of hospital inpatients. Few studies assessed the effectiveness of ice on closed soft-tissue injury, and there was no evidence of an optimal mode or duration of treatment. CONCLUSION: Many more high quality trials are needed to provide evidence-based guidelines in the treatment of acute soft-tissue injuries. PMID- 14754754 TI - Athletic injuries of the wrist and hand: part II: overuse injuries of the wrist and traumatic injuries to the hand. AB - Hand and wrist injuries in sports are some of the most common injuries reported. This review discusses common overuse injuries of the wrist including tendon injuries such as de Quervain's syndrome, subluxation of the extensor carpi ulnaris, and the common dorsal carpal impingement syndrome. The main focus of this section is the discussion of traumatic injuries to the hand in the athlete. Included is a discussion and review of fractures of the phalanges and metacarpals, common proximal interphalangeal joint injuries, and thumb carpal metacarpal and metacarpophalangeal joint injuries. Emphasis is placed on more common injuries seen regarding diagnosis, indications for non-operative versus operative treatment, and time to return to athletic competition. PMID- 14754755 TI - Protease-activated receptor-2-mediated inhibition for Ca2+ response to lipopolysaccharide in Guinea pig tracheal epithelial cells. AB - The protease-activated receptor-2 (PAR-2) has been implicated in airway inflammation. Here, we examined the interaction between PAR-2 and lipopolysaccharide (LPS), a major proinflammatory factor, using cultured guinea pig tracheal epithelial cells. In fura2-loaded cells, LPS (1 microg/ml) transiently increased intracellular Ca(2+) concentrations ([Ca(2+)]i), this effect being abolished by a Ca(2+) channel blocker, verapamil, and Ca(2+) removal. Prestimulation of PAR-2 with trypsin (0.1-1 U/ml) or an agonist peptide (SLIGRL-NH(2), 1 microM) for 60 min inhibited the LPS-induced [Ca(2+)]i increase. Such an inhibitory effect of trypsin was abolished by inhibitors of protein kinase C (PKC), chelerythrine and staurosporine. A PKC activator, phorbol 12,13 dibutylate, also reduced the LPS response. Trypsin also inhibited a transient increase in [Ca(2+)]i caused by a Ca(2+) channel opener, Bay K 8644. When the trypsin-pretreated cells were incubated in normal buffer for 10-60 min before LPS exposure, the effect of trypsin on the Ca(2+) response to LPS diminished in a time-dependent manner. Such a recovery was slowed by incubation with a protein phosphatase inhibitor, okadaic acid. Further, trypsin induced sustained activations of PKCalpha and -epsilon. Thus, PAR-2 stimulation reduced the epithelial cell response to LPS, probably through the inactivation of Ca(2+) channels via PKC-mediated phosphorylation. PMID- 14754756 TI - Ontogeny of poly(ADP-ribose) polymerase-1 in lung and developmental implications. AB - Poly(ADP-ribose) polymerase 1 (PARP-1) is the predominant NAD-dependent modifying enzyme in DNA repair, transcription, and apoptosis; its involvement in development has not been defined. Here, we report expression and cellular localization of PARP-1 in developing rat and human fetal lung, in vivo and in explant culture, and effects of inhibiting PARP-1 activity on lung surfactant protein (SP) expression. PARP-1 was expressed as 113-kD (p113) and 85-kD (p85) fragment in both rat and human lung. In rat lung, p113 content by Western was maximal at Embryonic Days 16-18, decreased sharply by Embryonic Day 20, and continued to decrease postnatally. p85 level was constant in the fetus and decreased postnatally. In human fetal lung, both PARP-1 mRNA expression and protein content changed little between 15 and 24 wk. Immunohistochemistry for PARP-1 in Embryonic Day 18 rat lung showed predominantly nuclear staining in most cells. In later gestation and postnatally, PARP-1 staining was primarily cytoplasmic and progressively restricted to a subset of cells, mainly bronchial epithelial and smooth muscle cells. Cell subfractionation showed that p113 localized to nucleus and p85 to cytoplasm. Inhibition of PARP-1 activity by 5 iodo-6-amino-1,2-benzopyrone in fetal rat lung explant culture did not affect SP A and -B mRNA, but significantly increased SP-C mRNA. These findings indicate that in lung (i) PARP-1 is abundantly expressed during fetal development; (ii) p113 and p85 levels are differentially regulated; (iii) PARP-1 undergoes complex developmental changes in cellular and subcellular expression, including extensive cytoplasmic localization; and (iv) inhibition of PARP-1 activity differentially affects expression of SPs. PMID- 14754757 TI - Hypoxia decreases expression of soluble guanylate cyclase in cultured rat pulmonary artery smooth muscle cells. AB - Nitric oxide (NO) has an important role in modulating the pulmonary vascular tone. NO acts, in part, by stimulating soluble guanylate cyclase (sGC) to synthesize the intracellular second messenger cyclic GMP. In vascular smooth muscle cells, sGC is a heterodimer composed of alpha1 and beta1 subunits. The objective of this study was to test whether oxygen concentration regulates sGC expression in cultured rat pulmonary artery smooth muscle cells (rPaSMC). rPaSMC were exposed to 0, 3, and 20% oxygen for 1-48 h, and sGC subunit mRNA levels were measured. Compared with rPaSMC exposed to 20% oxygen, sGC alpha1 and beta1 subunit mRNA levels were markedly decreased in rPaSMC exposed to 0% and 3% oxygen. The decrease in sGC subunit mRNA levels in hypoxic rPaSMC was detected as early as 6 h of exposure. Compared with rPaSMC exposed to 20% oxygen, exposure of rPaSMC to 3% oxygen progressively decreased sGC subunit protein levels at 24 and 48 h. There was also a 30% and 50% decrease in sGC enzyme activity in cells exposed to hypoxia for 24 and 48 h (P < 0.05 and P < 0.001, respectively, as compared with cells maintained in normoxia). These results demonstrate that hypoxia decreases sGC expression in cultured pulmonary artery smooth muscle cells and suggest that, in hypoxic vascular smooth muscle, decreased cyclic GMP synthesis may limit the vasodilator response to NO. PMID- 14754758 TI - Interleukin-1 receptor antagonist attenuates airway hyperresponsiveness following exposure to ozone. AB - The role of an interleukin (IL)-1 receptor antagonist (IL-1Ra) on the development of airway hyperresponsiveness (AHR) and airway inflammation following acute O(3) exposure in mice was investigated. Exposure of C57/BL6 mice to O(3) at a concentration of 2.0 ppm or filtered air for 3 h resulted in increases in airway responsiveness to inhaled methacholine (MCh) 8 and 16 h after the exposure, and an increase in neutrophils in the bronchoalveolar lavage (BAL) fluid. IL-1beta expression, assessed by gene microarray, was increased 2-fold 4 h after O(3) exposure, and returned to baseline levels by 24 h. Levels of IL-1beta in lung homogenates were also increased 8 h after O(3) exposure. Administration of (human) IL-1Ra before and after O(3) exposure prevented development of AHR and decreased BAL fluid neutrophilia. Increases in chemokine levels in lung homogenates, tumor necrosis factor-alpha, MIP-2, and keratinocyte chemoattractant following O(3) exposure were prevented by IL-1Ra. Inhalation of dexamethasone, an inhibitor of IL-1 production, blocked the development of AHR, BAL fluid neutrophilia, and decreased levels of IL-1 following O(3) exposure. In summary, acute exposure to O(3) induces AHR, neutrophilic inflammation, epithelial damage, and IL-1. An IL-1Ra effectively prevents the development of altered airway function, inflammation, and structural damage. PMID- 14754759 TI - Detection of airborne rhinovirus and its relation to outdoor air supply in office environments. AB - Rhinoviruses are major causes of morbidity in patients with respiratory diseases; however, their modes of transmission are controversial. We investigated detection of airborne rhinovirus in office environments by polymerase chain reaction technology and related detection to outdoor air supply rates. We sampled air from 9 A.M. to 5 P.M. each workday, with each sample run for 1 work week. We directly extracted RNA from the filters for nested reverse transcriptase-polymerase chain reaction analysis of rhinovirus. Nasal lavage samples from building occupants with upper respiratory infections were also collected. Indoor carbon dioxide (CO2 concentrations were recorded every 10 minutes as a surrogate for outdoor air supply. To increase the range of CO2 concentrations, we adjusted the outdoor air supply rates every 3 months. Generalized additive models demonstrated an association between the probability of detecting airborne rhinovirus and a weekly average CO2 concentration greater than approximately 100 ppm, after controlling for covariates. In addition, one rhinovirus from a nasal lavage contained an identical nucleic acid sequence similar to that in the building air collected during the same week. These results suggest that occupants in buildings with low outdoor air supply may have an increased risk of exposure to infectious droplet nuclei emanating from a fellow building occupant. PMID- 14754760 TI - Heterogeneous increase in CD34-positive alveolar capillaries in idiopathic pulmonary fibrosis. AB - To elucidate the apparent contradictions in vascular remodeling in the lungs of patients with idiopathic pulmonary fibrosis, we evaluated alveolar vascularity in relation to the various degrees of fibrosis in surgically biopsied lungs of usual interstitial pneumonia. Alveolar capillary endothelial cells were intensely immunoreactive with CD34 but not with von Willebrand factor. Vascular density, that is, the relative ratio of capillary area to total area of alveolar walls, was significantly higher at low grades of fibrosis than in control lungs, whereas vascular density gradually decreased as the degree of fibrosis increased and was lower than that of control lungs in the most extensively fibrotic lesions. No vessels were observed inside fibroblastic foci. The potent angiogenic factors vascular endothelial growth factor and interleukin-8 were abundantly produced by capillary endothelial cells and alveolar epithelial cells in highly vascularized alveolar walls. In contrast, venules with CD34-negative but von Willebrand factor positive endothelial cells localized in the center of the fibrotic lesions were slightly increased and identified as postcapillary venules by three-dimensional reconstructed images. These results indicate the presence of heterogeneous vascular remodeling in usual interstitial pneumonia. PMID- 14754761 TI - Is forced expiratory volume in one second the best measure of severity in childhood asthma? PMID- 14754762 TI - An investigation of inhaled ozone dose and the magnitude of airway inflammation in healthy adults. PMID- 14754763 TI - The evolution of airway function in early childhood following clinical diagnosis of cystic fibrosis. AB - This study aimed to investigate the evolution of airway function in infants newly diagnosed with cystic fibrosis (CF). FEV(0.5) was measured soon after diagnosis (median age of 28 weeks) and 6 months later in subjects with CF and on two occasions 6 months apart (median ages of 7.4 and 33.7 weeks) in healthy infants, using the raised-volume technique. Repeated measurements were successful in 34 CF and 32 healthy subjects. After adjustment for age, length, sex, and exposure to maternal smoking, mean FEV(0.5) was significantly lower in infants with CF both shortly after diagnosis and at the second test, with no significant difference in rate of increase in FEV(0.5) with growth between the two groups. When compared with published reference data, FEV(0.5) was reduced by an average of two z scores on both test occasions in those with CF, with 72% of individuals having an FEV(0.5) of less than 1.64 z-scores (i.e., less than the fifth percentile) on one or both test occasions. On longitudinal analysis, subjects with CF experienced a mean (95% confidence interval) reduction in FEV(0.5) of 20% (11, 28). Airway function is diminished soon after diagnosis in infants with CF and does not catch up during infancy and early childhood. These findings have important implications for early interventions in CF. PMID- 14754764 TI - Choose to study choice in addiction. PMID- 14754765 TI - Psychobiological mechanisms of resilience and vulnerability: implications for successful adaptation to extreme stress. AB - OBJECTIVE: Most research on the effects of severe psychological stress has focused on stress-related psychopathology. Here, the author develops psychobiological models of resilience to extreme stress. METHOD: An integrative model of resilience and vulnerability that encompasses the neurochemical response patterns to acute stress and the neural mechanisms mediating reward, fear conditioning and extinction, and social behavior is proposed. RESULTS: Eleven possible neurochemical, neuropeptide, and hormonal mediators of the psychobiological response to extreme stress were identified and related to resilience or vulnerability. The neural mechanisms of reward and motivation (hedonia, optimism, and learned helpfulness), fear responsiveness (effective behaviors despite fear), and adaptive social behavior (altruism, bonding, and teamwork) were found to be relevant to the character traits associated with resilience. CONCLUSIONS: The opportunity now exists to bring to bear the full power of advances in our understanding of the neurobiological basis of behavior to facilitate the discoveries needed to predict, prevent, and treat stress related psychopathology. PMID- 14754766 TI - Long-term lithium therapy for bipolar disorder: systematic review and meta analysis of randomized controlled trials. AB - OBJECTIVE: The authors sought to determine the efficacy and acceptability of lithium for relapse prevention in bipolar disorder. METHOD: A systematic review and meta-analysis of randomized controlled trials comparing lithium with placebo in the long-term treatment of bipolar disorders was conducted. Data were obtained from searching the registers of the Cochrane Collaboration; reviewing reference lists, journals, and conference abstracts; and contacting authors, experts, and pharmaceutical companies. Outcomes investigated included risk of relapse (manic, depressive, and total) as well as risk of specific adverse effects and total withdrawal rates. RESULTS: Five randomized controlled trials (770 participants) were included. Lithium was more effective than placebo in preventing all relapses (random effects relative risk=0.65, 95% CI=0.50 to 0.84) and manic relapses (relative risk=0.62, 95% CI=0.40 to 0.95). The protective effect of lithium on depressive relapses was smaller and was less robust (relative risk=0.72, 95% CI=0.49 to 1.07). CONCLUSIONS: Lithium treatment reduces the risk of relapse in bipolar disorder. The preventive effect is clear for manic episodes, although it is equivocal for depressive episodes. PMID- 14754767 TI - Drug dependence and addiction: neural substrates. PMID- 14754769 TI - Pure neuropsychiatric presentation of multiple sclerosis. PMID- 14754770 TI - Sigmund Freud, 1856-1939. PMID- 14754771 TI - The neural correlates of cue-induced craving in cocaine-dependent women. AB - OBJECTIVE: Drug use reminders are associated with localized changes in brain activity related to intense drug wanting or craving in cocaine-dependent men. While cocaine dependence is prevalent and disabling in women, and certain clinically relevant sex differences exist, there is an absence of knowledge related to the neural correlates of cocaine craving in cocaine-dependent women. METHOD: The differential neural response to imagery depicting cocaine use and neutral imagery was defined by using [15O]H2O positron emission tomography (PET) imaging in eight cocaine-dependent women. Results were compared with a matched group of eight cocaine-dependent men. RESULTS: Cocaine-related imagery was associated with relative increases in cocaine craving and increases in regional cerebral blood flow in the superior temporal gyrus, dorsal anterior and posterior cingulate cortex, nucleus accumbens area, and the central sulcus. Compared with the results of an identical PET study in matched cocaine-dependent men, conditioned cocaine craving in women was associated with less activation of the amygdala, insula, orbitofrontal cortex, and ventral cingulate cortex and greater activation of the central sulcus and widely distributed frontal cortical areas. CONCLUSIONS: These findings suggest the presence of sex differences in the functional anatomy of cue-induced cocaine craving associated with drug dependence. Such differences may reflect sex differences in conditioned associations to cocaine use, in affective and other corollaries of cocaine craving, or in their volitional regulation and may underlie apparent sex differences in the effects of cocaine abstinence and the expectations of treatment outcome. Some support for the need for sex-specific strategies for treatment of cocaine dependence is also furnished by the findings of this study. PMID- 14754772 TI - Partial recovery of brain metabolism in methamphetamine abusers after protracted abstinence. AB - OBJECTIVE: Methamphetamine is a highly addictive drug of abuse that is neurotoxic to dopamine terminals. The authors recently reported that decreases in dopamine transporters (used as markers of dopamine terminals) in the striatum of methamphetamine abusers recover with protracted abstinence and that relative to comparison subjects, recently detoxified methamphetamine abusers have lower metabolism in the striatum and thalamus. In this study, the authors assessed whether metabolism recovers with protracted abstinence. METHOD: Brain glucose metabolism was measured with positron emission tomography and [18F]fluorodeoxyglucose in five methamphetamine abusers who were evaluated after both a short (<6 months) and protracted (12-17 months) abstinence interval, eight methamphetamine abusers tested only after protracted abstinence, and 11 comparison subjects who were not drug users. RESULTS: Significantly greater thalamic, but not striatal, metabolism was seen following protracted abstinence relative to metabolism assessed after a short abstinence interval, and this increase was associated with improved performance in motor and verbal memory tests. Relative to the comparison subjects, the methamphetamine abusers tested after protracted abstinence had lower metabolism in the striatum (most accentuated in the caudate and nucleus accumbens) but not in the thalamus. CONCLUSIONS: The persistent decreases in striatal metabolism in methamphetamine abusers could reflect long-lasting changes in dopamine cell activity, and decreases in the nucleus accumbens could account for the persistence of amotivation and anhedonia in detoxified methamphetamine abusers. The recovery of thalamic metabolism could reflect adaptation responses to compensate for the dopamine deficits, and the associated improvement in neuropsychological performance further indicates its functional significance. These results suggest that while protracted abstinence may reverse some of the methamphetamine-induced alterations in brain function, other deficits persist. PMID- 14754773 TI - The effect of previous alcohol abuse on cognitive function in HIV infection. AB - OBJECTIVE: The authors' goal was to study the potential effect on cognitive function of an interaction of HIV infection and a history of alcohol abuse. METHOD: The subjects were 30 HIV-negative and 50 HIV-positive men with and without a past history of alcohol abuse. Thirty-three of the men (12 HIV negative and 21 HIV positive) had a past history of alcohol abuse, and 47 (18 HIV negative and 29 HIV positive) had never abused alcohol. Each subject's history of alcohol use was obtained by using a syndromal approach based on the Structured Clinical Interview for DSM-III-R and a quantitative approach. Each subject was given a battery of neuropsychological tests assessing verbal reasoning, reaction time, intelligence, memory, and dexterity. The subjects were then compared on a summary neuropsychological impairment rating. RESULTS: There were no significant differences in CD4 level, age, education, depression, anxiety, or other drug abuse history between the HIV-positive and HIV-negative groups with and without a history of alcohol abuse. Significant effects on cognitive function were found for past alcohol abuse and HIV infection, with significant interactions in verbal reasoning, auditory processing, and reaction time. This demonstrates that HIV infection and a history of alcohol abuse have independent effects on some aspects of higher cognitive function but may have synergistic effects on other cognitive domains. In the HIV-negative subjects there were no differences in cognitive function between subjects with and without a history of alcohol abuse. Among the HIV-positive subjects, those with a history of alcohol abuse performed more poorly on tests of verbal IQ, verbal reasoning, and reaction time. CONCLUSIONS: There are both additive and interactive effects of previous alcohol abuse and HIV infection on cognition. Individuals with a history of past alcohol abuse may be at greater risk for cognitive dysfunction in the context of HIV infection. PMID- 14754774 TI - Instability of symptoms in recurrent major depression: a prospective study. AB - OBJECTIVE: Recurrent episodes of major depressive disorder are reported to have similar or stable characteristics across episodes. However, symptoms appear to be moderately stable only in consecutive depressive episodes or if episode severity is considered. The authors prospectively studied major depressive episodes occurring within 2 years to determine whether symptoms in the second episode could be predicted on the basis of symptoms in the first. METHOD: Inpatients (N=78) with major depressive disorder were rated with the Hamilton Depression Rating Scale during two separate episodes. Patients had a baseline assessment at index hospitalization and follow-up visits at 3, 12, and 24 months after discharge. Information regarding the presence of a major depressive episode, its duration, and its severity was documented. Baseline and follow-up data were analyzed by using Pearson correlations with and without adjustments for severity of depression. RESULTS: Subtype of depression appeared not to be stable. The most robust, although still weak, correlations across episodes were for anxiety and suicidal behavior. Only modest correlations were identified for a few depressive symptoms. CONCLUSIONS: The lack of robust consistency of symptoms or depressive subtype across episodes is striking given the requirement of five of nine predetermined symptoms for depression, increasing the chances of finding an association. These findings suggest that there is a superfamily of mood disorders with pleomorphic manifestations across major depressive episodes within individual patients with unipolar depression. PMID- 14754775 TI - Cognitive function across manic or hypomanic, depressed, and euthymic states in bipolar disorder. AB - OBJECTIVE: The study aims were to address neuropsychological functioning across different states of bipolar illness and to determine relationships among clinical features, neuropsychological performance, and psychosocial functioning. METHOD: Several domains of cognitive function were examined in 30 depressed bipolar patients (DSM-IV criteria for major depression, Hamilton Depression Rating Scale score > or = 17), 34 manic or hypomanic bipolar patients (DSM-IV criteria for manic or hypomanic episode, Young Mania Rating Scale score > or = 12), and 44 euthymic bipolar patients (6 months of remission, Hamilton depression scale score < or = 8, and Young Mania Rating Scale score < or = 6). The comparison group consisted of 30 healthy subjects without history of neurological or psychiatric disorders. A neuropsychological battery assessed executive function, attention, and verbal and visual memory. RESULTS: The three groups showed cognitive dysfunction in verbal memory and frontal executive tasks in relation to the comparison group. Low neuropsychological performance was associated with poor functional outcome. Impairment of verbal memory was related to the duration of illness and the numbers of previous manic episodes, hospitalizations, and suicide attempts. CONCLUSIONS: A poorer performance was observed in all bipolar groups regarding executive function and verbal memory in relation to the healthy comparison subjects. These cognitive difficulties, especially related to verbal memory, may help explain the impairment regarding daily functioning, even during remission. Further studies should focus on testing, whether optimizing prophylactic pharmacological treatment and psychoeducation might reduce cognitive impairment, and whether bipolar patients would benefit from neuropsychological rehabilitation in order to reduce the impact of cognitive impairment in their overall functioning. PMID- 14754776 TI - The association between major depression and levels of soluble intercellular adhesion molecule 1, interleukin-6, and C-reactive protein in patients with recent acute coronary syndromes. AB - OBJECTIVE: This study was conducted to determine whether or not depression is associated with higher levels of inflammatory markers in patients recovering from acute coronary syndromes. METHOD: Plasma levels of soluble intercellular adhesion molecule 1 (sICAM-1) and interleukin-6 (IL-6) and the serum level of C-reactive protein were measured in 481 patients 2 months after hospitalization for acute coronary syndromes. Diagnosis of major depression was based on the Structured Clinical Interview for DSM-IV. RESULTS: Depressed patients showed significantly higher sICAM-1 levels, a difference that remained significant after adjustment for potential confounders (gender, smoking, presence of metabolic syndrome). Although there was no significant association between depression and IL-6, there was an interaction between depression and statin therapy for levels of C-reactive protein. Depressed patients not taking statins had markedly higher C-reactive protein levels than did nondepressed patients. There was no relationship with depression in those receiving statins. CONCLUSIONS: These results suggest chronic endothelial activation among depressed patients after acute coronary syndromes. Further research is needed to determine whether or not higher levels of sICAM-1 may identify a subgroup of depressed patients at particularly high risk for cardiac events among patients with established coronary artery disease or among those without previous coronary artery disease. PMID- 14754777 TI - Distress and depression in men who have sex with men: the Urban Men's Health Study. AB - OBJECTIVE: This study estimates the prevalence of depression and describes the correlates and independent associations of distress and depression among U.S. men who have sex with men. METHOD: A household-based probability sample of men who have sex with men (N=2,881) was interviewed between 1996 and 1998 in four large American cities. With cutoff points of 15 and 22 for the Center for Epidemiological Studies Depression Scale, individual correlates and predictors of distress and depression were examined, and multinomial logistic regression was performed. RESULTS: The 7-day prevalence of depression in men who have sex with men was 17.2%, higher than in adult U.S. men in general. Both distress and depression were associated with lack of a domestic partner; not identifying as gay, queer, or homosexual; experiencing multiple episodes of antigay violence in the previous 5 years; and very high levels of community alienation. Distress was also associated with being of other than Asian/Pacific Islander ethnicity and experiencing early antigay harassment. Depression was also associated with histories of attempted suicide, child abuse, and recent sexual dysfunction. Being HIV positive was correlated with distress and depression but not significantly when demographic characteristics, developmental history, substance use, sexual behavior, and current social context were controlled by logistic regression. CONCLUSIONS: Rates of distress and depression are high in men who have sex with men. These high rates have important public health ramifications. The predictors of distress and depression suggest prevention efforts that might be effective when aimed at men who have sex with men. PMID- 14754778 TI - Brain activation measured with fMRI during a mental arithmetic task in schizophrenia and major depression. AB - OBJECTIVE: The authors used functional magnetic resonance imaging (fMRI) to investigate brain activation in patients with schizophrenia and major depression while they performed two tasks-a vigilance task and a mental arithmetic task-that differed in cognitive complexity. METHOD: In the vigilance task, the participants had to press a response button whenever a specific number was seen on a screen inside the MR scanner. In the mental arithmetic task, the participants had to add two consecutive numbers and press the response button whenever the sum was 10. fMRI was performed with a 1.5-T MR scanner. Twelve patients with recurrent nonpsychotic unipolar major depression, 12 patients with schizophrenia, and 12 healthy comparison subjects were included in the study. RESULTS: Performance data showed that the patients were impaired relative to the comparison subjects and showed no difference in performance between the patient groups. The patients with schizophrenia, but not those with major depression, had less activation in prefrontal brain regions, relative to the comparison participants. However, subtracting brain activation during the vigilance task from activation during the mental arithmetic task showed that the schizophrenia patients had activation in parietal areas. CONCLUSIONS: A double dissociation of parietal and frontal lobe activation was found for the schizophrenia patients and the depression patients. The greater parietal lobe activation in the patients with schizophrenia may reflect a compensatory strategy for the failure to recruit cognitive processes that involve frontal lobe areas when solving a mental arithmetic task. PMID- 14754779 TI - Mismatch negativity responses in schizophrenia: a combined fMRI and whole-head MEG study. AB - OBJECTIVE: Mismatch negativity is an event-related brain response sensitive to deviations within a sequence of repetitive auditory stimuli. It is thought to reflect short-term sensory memory and is independent of higher-level cognitive processes. Mismatch negativity response is diminished in patients with schizophrenia. Little is known about the mechanisms of this decreased response, the contribution of the different hemispheres, and its locus of generation. METHOD: Patients with schizophrenia (N=12) and matched comparison subjects (N=12) were studied. A novel design to measure mismatch negativity responses to deviant auditory stimuli was generated by using the switching noises from the functional magnetic resonance imaging (fMRI) scanner, thus avoiding any interfering background sound. Stimuli included deviants of amplitude (9 dB lower) and duration (76 msec shorter) presented in a random sequence. The scanner noise was recorded and applied to the same subjects in a whole-head magnetoencephalography (MEG) device. Neuromagnetic and hemodynamic responses to the identical stimuli were compared between the patients and comparison subjects. RESULTS: As expected, neuromagnetic mismatch fields were smaller in the patient group. More specifically, a lateralization to the right for duration deviance was only found in comparison subjects. For the relative amplitude of the blood-oxygen-level dependent signal (measured with fMRI), differences emerged in the secondary (planum temporale), but not primary (Heschl's gyrus), auditory cortex. Duration deviants achieved a right hemispheric advantage only in the comparison group. A significantly stronger lateralization to the left was found for the deviant amplitude stimuli in the patients. CONCLUSIONS: The data support the view of altered hemispheric interactions in the formation of the short-term memory traces necessary for the integration of auditory stimuli. This process is predominantly mediated by the planum temporale (secondary auditory cortex). Altered interaction of regions within the superior temporal plane and across hemispheres could be in part responsible for language-mediated cognitive (e.g., verbal memory) and psychopathological (hallucinations, formal thought disorder) symptoms in schizophrenia. PMID- 14754780 TI - Abnormal glucose metabolism in the mediodorsal nucleus of the thalamus in schizophrenia. AB - OBJECTIVE: Three thalamic nuclei--the mediodorsal nucleus, pulvinar, and centromedian nucleus--each have unique reciprocal circuitry with cortical and subcortical areas known to be affected in schizophrenia. To determine if the disorder is also associated with dysfunction in the mediodorsal nucleus, pulvinar, and centromedian nucleus, relative glucose metabolism in these regions was measured in a large group of unmedicated patients with schizophrenia. METHOD: [18F]-deoxyglucose positron emission tomography (PET) and matching T1-weighted magnetic resonance imaging (MRI) scans were obtained for 41 unmedicated patients with schizophrenia and 60 age- and sex-matched healthy subjects. The PET and MRI images for each subject were coregistered, and the whole thalamus, mediodorsal nucleus, pulvinar, and centromedian nucleus were traced on the MRI image. Relative glucose metabolism in these regions was assessed. RESULTS: Patients with schizophrenia showed significantly lower relative glucose metabolism in the mediodorsal nucleus and the centromedian nucleus and significantly higher relative glucose metabolism in the pulvinar, compared with the healthy subjects. Lower relative glucose metabolism in the total thalamus, mediodorsal nucleus, and pulvinar was associated with greater overall clinical symptoms as measured by the Brief Psychiatric Rating Scale. Lower relative glucose metabolism in the pulvinar was associated with more hallucinations and more positive symptoms, while lower relative glucose metabolism in the mediodorsal nucleus was associated with more negative symptoms. CONCLUSIONS: The findings suggest that patients with schizophrenia exhibit dysfunction in thalamic subdivisions with distinct cortical connections and that these thalamic subdivisions have specific associations with clinical symptoms. PMID- 14754781 TI - Neurobiology of smooth pursuit eye movement deficits in schizophrenia: an fMRI study. AB - OBJECTIVE: Smooth pursuit eye movements are abnormal in patients with schizophrenia. The investigators used functional magnetic resonance imaging (fMRI) to compare brain hemodynamic response during a smooth pursuit eye movement task in patients with schizophrenia and healthy comparison subjects. METHOD: Fourteen patients with schizophrenia and 14 healthy comparison subjects performed a smooth pursuit eye movement task while undergoing 1.5-T fMRI. Echo-planar images from the blocked design were evaluated with both a whole-brain random effects analysis and a region-of-interest analysis. Smooth pursuit deficits were assessed outside the fMRI apparatus by using infrared oculography and were assessed during scanning by evaluating echo-planar time-series data from the eyes. RESULTS: Compared to the healthy subjects, the patients with schizophrenia exhibited greater activity in both posterior hippocampi and the right fusiform gyrus during smooth pursuit eye movements. The region-of-interest analysis revealed subtle activity deficits in frontal and occipital regions in the patients with schizophrenia. Smooth pursuit deficits in the subjects with schizophrenia included lower gain (eye velocity relative to target velocity) and a higher percentage of total eye movements due to anticipatory saccades, compared with the healthy subjects. CONCLUSIONS: The observed group differences are consistent with evidence for diminished inhibitory function in the hippocampus as well as for a disturbance in a frontotemporal network subserving smooth pursuit eye movements in schizophrenia. PMID- 14754782 TI - Morphometric assessment of the heteromodal association cortex in schizophrenia. AB - OBJECTIVE: The heteromodal association cortex has been hypothesized to be selectively involved in the pathophysiology of schizophrenia. To test this hypothesis, the authors measured prefrontal, inferior parietal, and superior temporal gyrus volumes and examined the pattern of connections among these regions. METHOD: Forty-four patients with schizophrenia or schizoaffective disorder and 34 healthy comparison subjects were included in the study. A spoiled gradient recall acquisition in the steady-state three-dimensional magnetic resonance imaging sequence was used for morphometric assessment of the heteromodal association cortex. RESULTS: Patients with schizophrenia had significantly smaller inferior prefrontal region volumes and significant reversal of the normal asymmetry of the inferior parietal cortex. No significant group differences were found in superior temporal gyrus volume. The groups differed significantly in the correlation between inferior prefrontal region volumes and angular gyrus volumes. CONCLUSIONS: The results suggest that patients with schizophrenia may be characterized by selective abnormalities of the heteromodal regions involved in the neuroanatomy of language. PMID- 14754783 TI - Randomized clinical trial of supervised tapering and cognitive behavior therapy to facilitate benzodiazepine discontinuation in older adults with chronic insomnia. AB - OBJECTIVE: This study evaluated the effectiveness of a supervised benzodiazepine taper, singly and combined with cognitive behavior therapy, for benzodiazepine discontinuation in older adults with chronic insomnia. METHOD: Seventy-six older adult outpatients (38 women, 38 men; mean age of 62.5 years) with chronic insomnia and prolonged use (mean duration of 19.3 years) of benzodiazepine medication for sleep were randomly assigned for a 10-week intervention consisting of a supervised benzodiazepine withdrawal program (N=25), cognitive behavior therapy for insomnia (N=24), or supervised withdrawal plus cognitive behavior therapy (N=27). Follow-up assessments were conducted at 3 and 12 months. The main outcome measures were benzodiazepine use, sleep parameters, and anxiety and depressive symptoms. RESULTS: All three interventions produced significant reductions in both the quantity (90% reduction) and frequency (80% reduction) of benzodiazepine use, and 63% of the patients were drug-free within an average of 7 weeks. More patients who received medication taper plus cognitive behavior therapy (85%) were benzodiazepine-free after the initial intervention, compared to those who received medication taper alone (48%) and cognitive behavior therapy alone (54%). The patients in the two groups that received cognitive behavior therapy perceived greater subjective sleep improvements than those who received medication taper alone. Polysomnographic data showed an increase in the amount of time spent in stages 3 and 4 sleep and REM sleep and a decrease in total sleep time across all three conditions from baseline to posttreatment. Initial benzodiazepine reductions were well maintained up to the 12-month follow-up, and sleep improvements became more noticeable over this period. No significant withdrawal symptoms or adverse events were associated with benzodiazepine tapering. CONCLUSIONS: A structured, time-limited intervention is effective in assisting chronic users of benzodiazepine medication to discontinue or reduce their use of medication. The addition of cognitive behavior therapy alleviates insomnia, but sleep improvements may become noticeable only after several months of benzodiazepine abstinence. PMID- 14754784 TI - Continuous or intermittent dosing with sertraline for patients with severe premenstrual syndrome or premenstrual dysphoric disorder. AB - OBJECTIVE: The authors compared the efficacy and acceptability of continuous versus intermittent treatment with a selective serotonin reuptake inhibitor in women with severe premenstrual syndrome and determined the effects of postmenstrual symptom severity and depression history as covariates of the treatment response. METHOD: Patients who met symptom criteria and reported impaired functioning after three screening cycles were randomly assigned to three cycles of double-blind, placebo-controlled treatment with continuous (full-cycle dosing) or intermittent (luteal-phase dosing) sertraline. The design was stratified for severity of postmenstrual symptoms and history of major depression. Flexible sertraline dose was 50-100 mg/day. Outcome measures were the Daily Symptom Rating Form score and patient global ratings of functioning. RESULTS: Both sertraline groups improved significantly more than the placebo group as assessed by total premenstrual Daily Symptom Rating Form scores for 3 treatment months. Daily Symptom Rating Form factors that were significantly more improved in the sertraline groups were mood and physical symptoms. Sertraline improvement occurred swiftly in the first month of treatment. Gradual placebo improvement was similar to sertraline in the third month. Subjects with higher postmenstrual symptoms before treatment remained more symptomatic regardless of the dosing regimen. A history of major depression was not associated with treatment response. More sertraline-treated subjects reported improved functioning in the domains of family relationships, social activities, and sexual activity. CONCLUSIONS: Premenstrual dosing does not differ from continuous dosing with sertraline in premenstrual syndrome treatment. Higher levels of postmenstrual symptoms limit treatment response and are important to define in treatment of premenstrual syndrome. PMID- 14754785 TI - Geriatric psychiatry versus general psychiatry inpatient treatment of the elderly. AB - OBJECTIVE: The authors compared the clinical treatment given older psychiatric inpatients on a geriatric psychiatry unit and a general psychiatry unit. METHOD: The charts of 50 randomly selected general psychiatry inpatients over the age of 65 years and 50 inpatients from the geriatric psychiatry unit who were matched for age, gender, and primary diagnosis were reviewed. RESULTS: Significantly greater percentages of older inpatients treated on the geriatric psychiatry unit received complete organic medical workups, structured cognitive assessment, aging sensitive aftercare referral, and monitoring of psychopharmacological side effects and blood levels than comparable patients on a general psychiatry unit. CONCLUSIONS: Geriatric psychiatry subspecialty inpatient care appears to be associated with distinct clinically relevant assessment and treatment advantages. Continuing geropsychiatric education of general psychiatrists is indicated. PMID- 14754786 TI - Are oppositional defiant and conduct disorder symptoms normative behaviors in preschoolers? A comparison of referred and nonreferred children. AB - OBJECTIVE: The authors' goal was to test the hypothesis that DSM-IV symptoms of oppositional defiant disorder and conduct disorder can be validly applied to preschoolers. METHOD: Using a semistructured diagnostic interview, they assessed rates of symptoms of DSM-IV oppositional defiant and conduct disorders in 2.5-5.5 year-old children who were referred to a psychiatry clinic and a comparison group of nonreferred children. RESULTS: Clinically referred preschool children had significantly higher rates of oppositional defiant and conduct disorder symptoms than nonreferred children. Among nonreferred preschoolers, rates of all oppositional defiant and conduct disorder symptoms were at or below 8%. CONCLUSIONS: DSM-IV symptoms of oppositional defiant and conduct disorders distinguish referred from nonreferred preschool children in a pattern consistent with that seen in older children. Preschool children who are not seeking mental health services do not have high rates of disruptive behavior problems. The DSM IV nosology appears to be a valid diagnostic system for discriminating between typical and atypical disruptive behaviors in preschool children. PMID- 14754787 TI - Catechol O-methyltransferase Val158Met polymorphism in schizophrenia: differential effects of Val and Met alleles on cognitive stability and flexibility. AB - OBJECTIVE: The catechol O-methyltransferase (COMT) Val158Met polymorphism has been associated with cognitive and behavioral phenotypes in schizophrenia. Whether COMT genotype is beneficial may depend on phenotype definition. The authors examined the effects of COMT genotype on a task that distinguishes imitation from reversal learning. They hypothesized that the Val and Met alleles would be associated with deficits in imitation learning and reversal learning, respectively. METHOD: Twenty-six patients with schizophrenia and schizoaffective disorder completed a task requiring alternation between imitation and reversal rules. RESULTS: Met homozygotes showed better acquisition of the imitation rule but greater deficit shifting from imitation to reversal. Val homozygotes had poorer imitation performance and slower reaction times. CONCLUSIONS: The Met allele, by increasing tonic dopamine, may promote cognitive stability but limit cognitive flexibility. PMID- 14754788 TI - A prospective study of impairment in glucose control caused by clozapine without changes in insulin resistance. AB - OBJECTIVE: This prospective study examines the effect of clozapine on glucose control and insulin sensitivity. METHOD: Glucose homeostasis was measured in nine female and 11 male patients with schizophrenia (mean age=30.5 years, SD=7.4) before clozapine treatment and after a mean of 2.5 months (SD=0.95) of clozapine treatment. Oral glucose tolerance and insulin levels were measured. Insulin resistance level was measured by the homeostasis model assessment. RESULTS: Eleven (55%) of the patients developed abnormal glucose control; the mean age of these patients was 30.2 (SD=7.1), and five were women. Patients' insulin resistance at baseline (mean insulin resistance level=3.88, SD=2.93) was unaffected by clozapine. Mean fasting and 2-hour glucose levels significantly increased by 0.55 mmol/liter and 1.4 mmol/liter, respectively. There was no correlation between change in body mass index and change in fasting glucose levels. CONCLUSIONS: Clozapine impairs glucose control within 4 months of treatment, independent of changes in insulin sensitivity and body mass index. PMID- 14754790 TI - Increased brain GABA concentrations following acute administration of a selective serotonin reuptake inhibitor. AB - OBJECTIVE: The authors used magnetic resonance spectroscopy (MRS) to assess the effect of acute administration of the selective serotonin reuptake inhibitor (SSRI) citalopram on cortical levels of gamma-aminobutyric acid (GABA). METHOD: Ten healthy volunteers received either intravenous citalopram (10 mg) or saline in a randomized, double-blind, crossover design. The occipital GABA/creatine ratio was measured with a proton MR spectral editing technique. RESULTS: In comparison with saline, citalopram produced a mean increase of 35% in relative brain GABA concentration in the occipital cortex. CONCLUSIONS: These findings extend previous work showing that SSRI treatment increases cortical GABA in depressed patients and suggest that this results from an action of SSRIs on GABA neurons rather than as a secondary consequence of mood improvement. PMID- 14754789 TI - Do clozapine and risperidone affect social competence and problem solving? AB - OBJECTIVE: The purpose of this investigation was to evaluate the effects of clozapine and risperidone on social skill and problem solving in patients with schizophrenia. METHOD: The Wisconsin Card Sorting Test and the Maryland Assessment of Social Competence were administered at baseline, week 17, and week 29 of a multisite clinical trial. RESULTS: Despite evidence of clinical improvement with both medications, there was virtually no medication effect on either social competence or problem solving. CONCLUSIONS: These findings underscore the circumscribed nature of symptomatic improvement in the broader spectrum of clinical outcomes and suggest that new-generation medications may not be expected to produce substantial changes in social role functioning or social problem-solving capacity in the community. The generalizability of the findings should be viewed cautiously because of the low power of this trial, and replication is warranted. PMID- 14754791 TI - Response to tricyclic antidepressants: independent of gender? AB - OBJECTIVE: The authors examined gender differences in response to tricyclic antidepressants. METHOD: A total of 30 randomized, placebo-controlled trials that included 3,886 patients (1,555 men and 2,331 women), submitted between 1979 and 1991 in order to obtain marketing authorization, were reviewed. Gender differences in response to treatment were tested in various multiple regression models using a variety of response definitions. RESULTS: Different response definitions all pointed to no gender difference in the efficacy of tricyclic antidepressants. The estimated effect size was similar for women younger and older than age 50 and for men. CONCLUSIONS: Tricyclic antidepressant response is independent of gender. PMID- 14754792 TI - Aripiprazole and Parkinson's disease psychosis. PMID- 14754793 TI - Pisa syndrome and atypical antipsychotics. PMID- 14754794 TI - Worsening of hyponatremia with electrolyte-containing beverage. PMID- 14754795 TI - Suicide attempt due to a misunderstood HIPAA notice. PMID- 14754796 TI - Gamma-hydroxybutyrate dependence with social phobia. PMID- 14754797 TI - On biology, phenomenology, and pharmacology in schizophrenia. PMID- 14754798 TI - On biology, phenomenology, and pharmacology in schizophrenia. PMID- 14754799 TI - On biology, phenomenology, and pharmacology in schizophrenia. PMID- 14754801 TI - On biology, phenomenology, and pharmacology in schizophrenia. PMID- 14754811 TI - Introduction: Professor Channi Kumar (1938-2000). PMID- 14754812 TI - Aims, measures, study sites and participant samples of the Transcultural Study of Postnatal Depression. AB - BACKGROUND: Although postnatal depression is a common condition, with adverse effects on the child, mother and partner, there has been no research to date linking analyses of its origins and consequences with studies of service provision in different national health and social systems. AIMS: The Transcultural Study of Postnatal Depression (TCS-PND) was set up to develop a set of instruments to facilitate such research. METHOD: Seven research instruments were piloted in studies carried out in ten centres in eight countries. A qualitative study looked at the cross-cultural equivalence of the concept of postnatal depression. RESULTS: The results are reported in the individual papers in this supplement. CONCLUSIONS: Reliable and culturally valid measures are now available for future comparative research into postnatal depression within different health systems and countries. PMID- 14754813 TI - Postnatal depression across countries and cultures: a qualitative study. AB - BACKGROUND: Postnatal depression seems to be a universal condition with similar rates in different countries. However, anthropologists question the cross cultural equivalence of depression, particularly at a life stage so influenced by cultural factors. AIMS: To develop a qualitative method to explore whether postnatal depression is universally recognised, attributed and described and to enquire into people's perceptions of remedies and services for morbid states of unhappiness within the context of local services. METHOD: The study took place in 15 centres in 11 countries and drew on three groups of informants: focus groups with new mothers, interviews with fathers and grandmothers, and interviews with health professionals. Textual analysis of these three groups was conducted separately in each centre and emergent themes compared across centres. RESULTS: All centres described morbid unhappiness after childbirth comparable to postnatal depression but not all saw this as an illness remediable by health interventions. CONCLUSIONS: Although the findings of this study support the universality of a morbid state of unhappiness following childbirth, they also support concerns about the cross-cultural equivalence of postnatal depression as an illness requiring the intervention of health professionals; this has implications for future research. PMID- 14754814 TI - Adaptation of the structured clinical interview for DSM-IV disorders for assessing depression in women during pregnancy and post-partum across countries and cultures. AB - BACKGROUND: To date, no study has used standardised diagnostic assessment procedures to determine whether rates of perinatal depression vary across cultures. AIMS: To adapt the Structured Clinical Interview for DSM-IV Disorders (SCID) for assessing depression and other non-psychotic psychiatric illness perinatally and to pilot the instrument in different centres and cultures. METHOD: Assessments using the adapted SCID and the Edinburgh Postnatal Depression Scale were conducted during the third trimester of pregnancy and at 6 months postpartum with 296 women from ten sites in eight countries. Point prevalence rates during pregnancy and the postnatal period and adjusted 6-month period prevalence rates were computed for caseness, depression and major depression. RESULTS: The third trimester and 6-month point prevalence rates for perinatal depression were 6.9% and 8.0%, respectively. Postnatal 6-month period prevalence rates for perinatal depression ranged from 2.1% to 31.6% across centres and there were significant differences in these rates between centres. CONCLUSIONS: Study findings suggest that the SCID was successfully adapted for this context. Further research on determinants of differences in prevalence of depression across cultures is needed. PMID- 14754815 TI - Contextual Assessment of the Maternity Experience: development of an instrument for cross-cultural research. AB - BACKGROUND: There is evidence that stressors may trigger the onset of a depressive episode in vulnerable women. A new UK interview measure, the Contextual Assessment of the Maternity Experience (CAME), was designed to assess major risk factors for emotional disturbances, especially depression, during pregnancy and post-partum. AIMS: With in the context of a cross-cultural study, to establish the usefulness of the CAME, and to test expected associations of the measure with characteristics of the social context and with major or minor depression. METHOD: The CAME was administered antenatally and postnatally in ten study sites, respectively to 296 and 249 women. Affective disorder throughout pregnancy and up to 6 months postnatally was assessed by means of the Structured Clinical Interview for DSM-IVAxis I Disorders. RESULTS: Adversity, poor relationship with either a partner or a confidant, and negative feelings about the pregnancy all predicted onset of depression during the perinatal period. CONCLUSIONS: The CAME was able to assess major domains relevant to the psychosocial context of the maternity experience in different cultures. Overall, the instrument showed acceptable psychometric properties in its first use in different cultural settings. PMID- 14754816 TI - Maternal attachment style and depression associated with childbirth: preliminary results from a European and US cross-cultural study. AB - BACKGROUND: Insecure attachment style relates to major depression in women, but its relationship to depression associated with childbirth is largely unknown. A new UK-designed measure, the Attachment Style Interview (ASI), has potential for cross-cultural use as a risk marker for maternal disorder. AIMS: To establish there liability of the ASI across centres, its stability over a 9-month period, and its associations with social context and major or minor depression. METHOD: The ASI was used by nine centres antenatally on 204 women, with 174 followed up 6 months postnatally. Interrater reliability was tested and the ASI was repeated on a subset of 96 women. Affective disorder was assessed by means of the Structured Clinical Interview for DSM-IV. RESULTS: Satisfactory interrater reliability was achieved with relatively high stability rates at follow-up. Insecure attachment related to lower social class position and more negative social context. Specific associations of avoidant attachment style (angry-dismissive or withdrawn) with antenatal disorder, and anxious style (enmeshed or fearful) with postnatal disorder were found. CONCLUSIONS: The ASI can be used reliably in European and US centres as a measure for risk associated with childbirth. Its use will contribute to theoretically under pinned preventive action for disorders associated with childbirth. PMID- 14754817 TI - Measurement of mother-infant interactions and the home environment in a European setting: preliminary results from a cross-cultural study. AB - BACKGROUND: Infant development is adversely affected in the context of postnatal depression. This relationship may be mediated by both the nature of early mother infant interactions and the quality of the home environment. AIM: To establish the usefulness of the Global Ratings Scales of Mother-Infant Interaction and the Infant-Toddler version of the Home Observation for the Measurement of the Environment (IT-HOME), and to test expected associations of the measures with characteristics of the social context and with major or minor depression. METHOD: Both assessments were administered postnatally in four European centres; 144 mothers were assessed with the Global Ratings Scales and 114 with the IT-HOME. Affective disorder was assessed by means of the Structured Clinical Interview for DSM-IV Disorders. RESULTS: Analyses of mother-infant interaction indicated no main effect for depression but maternal sensitivity to infant behaviour was associated with better infant communication, especially for women who were not depressed. Poor overall emotional support also reduced sensitivity scores. Poor support was also related to poorer IT-HOME scores, but there was no effect of depression. CONCLUSIONS: The Global Ratings Scales were effectively applied but there was less evidence of the usefulness of the IT-HOME. PMID- 14754818 TI - Health services research into postnatal depression: results from a preliminary cross-cultural study. AB - BACKGROUND: Little is known about the availability and uptake of health and welfare services by women with postnatal depression in different countries. AIMS: Within the context of a cross-cultural research study, to develop and test methods for undertaking quantitative health services research in postnatal depression. METHOD: Interviews with service planners and the collation of key health indicators were used to obtain a profile of service availability and provision. A service use questionnaire was developed and administered to a pilot sample in a number of European study centres. RESULTS: Marked differences in service access and use were observed between the centres, including postnatal nursing care and contacts with primary care services. Rates of use of specialist services were generally low. Common barriers to access to care included perceived service quality and responsiveness. On the basis of the pilot work, a postnatal depression version of the Service Receipt Inventory was revised and finalised. CONCLUSIONS: This preliminary study demonstrated the methodological feasibility of describing and quantifying service use, highlighted the varied and often limited use of care in this population, and indicated the need for an improved understanding of the resource needs and implications of postnatal depression. PMID- 14754819 TI - Social psychiatry and the human genome: contextualising heritability. PMID- 14754820 TI - Liquid ecstasy: a new kid on the dance floor. PMID- 14754821 TI - Culture and insight revisited. PMID- 14754822 TI - Causal association between cannabis and psychosis: examination of the evidence. AB - BACKGROUND: Controversy remains as to whether cannabis acts as a causal risk factor for schizophrenia or other functional psychotic illnesses. AIMS: To examine critically the evidence that cannabis causes psychosis using established criteria of causality. METHOD: We identified five studies that included a well defined sample drawn from population-based registers or cohorts and used prospective measures of cannabis use and adult psychosis. RESULTS: On an individual level, cannabis use confers an overall twofold increase in the relative risk for later schizophrenia. At the population level, elimination of cannabis use would reduce the incidence of schizophrenia by approximately 8%, assuming a causal relationship. Cannabis use appears to be neither a sufficient nor a necessary cause for psychosis. It is a component cause, part of a complex constellation of factors leading to psychosis. CONCLUSIONS: Cases of psychotic disorder could be prevented by discouraging cannabis use among vulnerable youths. Research is needed to understand the mechanisms by which cannabis causes psychosis. PMID- 14754823 TI - Predictors of antisocial personality. Continuities from childhood to adult life. AB - BACKGROUND: Antisocial behaviour in adult life has its roots in childhood. AIMS: To explore the independent and joint effects of childhood characteristics on the persistence of antisocial behaviour into adult life. METHOD: A clinical sample of twins who were systematically ascertained in childhood was followed up 10-25 years later. A total of 225 twins were interviewed regarding childhood and adult psychiatric disorder, psychosocial functioning, and psychosocial and cognitive risk factors. RESULTS: In univariate analyses, childhood hyperactivity and conduct disorder showed equally strong prediction of antisocial personality disorder (ASPD) and criminality in early and mid-adult life. Lower IQ and reading problems were most prominent in their relationships with childhood and adolescent antisocial behaviour. In multivariate modelling childhood conduct disorder and hyperactivity predicted adult ASPD even when intervening risk factors were accounted for. The number of hyperactive and conduct symptoms also predicted adult outcome. CONCLUSIONS: Childhood disruptive behaviour has powerful long-term effects on adult antisocial outcomes, which continue into middle adulthood. The importance of number of symptoms, the presence of disruptive disorder, and intermediate experiences highlight three areas where interventions might be targeted. PMID- 14754824 TI - Language activation in monozygotic twins discordant for schizophrenia. AB - BACKGROUND: In previous functional magnetic resonance imaging (fMRI) studies, participants with schizophrenia showed decreased language lateralisation, resulting from increased activation of the right hemisphere compared with controls. AIM: To determine whether decreased lateralisation and increased right cerebral language activation constitute genetic predispositions for schizophrenia. METHOD: Language activation was measured using fMRI in 12 right handed monozygotic twin pairs discordant for schizophrenia and 12 healthy right handed monozygotic twin pairs who were twin pairs who were matched for gender, age and education. RESULTS: Language lateralisation was decreased in discordant twin pairs compared with the healthy twin pairs. The groups did not differ in activation of the language-related areas of the left hemisphere, but language related activation in the right hemisphere was activation in the significantly higher in the discordant twin pairs than in the healthy pairs. Within the discordant twin pairs, language lateralisation was not significantly different between patients with schizophrenia and their co-twins. CONCLUSIONS: Decreased language lateralisation may constitute a genetic predisposition for schizophrenia. PMID- 14754825 TI - Salivary cortisol response to awakening in chronic fatigue syndrome. AB - BACKGROUND: There is accumulating evidence of hypothalamic-pituitary-adrenal (HPA) axis disturbances in chronic fatigue syndrome (CFS). The salivary cortisol response to awakening has been described recently as a non-invasive test of the capacity of the HPA axis to respond to stress. The results of this test correlate closely with those of more invasive dynamic tests reported in the literature; furthermore, it can be undertaken in a naturalistic setting. AIMS: To assess the HPA axis using the salivary cortisol response to awakening in CFS. METHOD: We measured salivary cortisol upon awakening and 10, 20, 30 and 60 min afterwards in 56 patients with CFS and 35 healthy volunteers. RESULTS: Patients had a lower cortisol response to awakening, measured by the area under the curve. CONCLUSIONS: This naturalistic test of the HPA axis response to stress showed impaired HPA axis function in CFS. PMID- 14754826 TI - Patient education to encourage graded exercise in chronic fatigue syndrome. 2 year follow-up of randomised controlled trial. AB - BACKGROUND: An earlier trial demonstrated good outcomes after 1 year for patients with chronic fatigue syndrome (CFS) who received an educational intervention designed to encourage graded activity. AIMS: To determine 2-year outcomes for the same treated patients and the response to treatment of patients formerly in the control condition. METHOD: Patients in the treatment groups (n=114) were followed up at 2 years; 32 patients from the control group were offered the intervention after 1 year and were assessed 1 year later. Assessments were the self-rated measures used in the original trial. RESULTS: At 2 years 63 of the treated patients (55%) no longer fulfilled trial criteria for CFS compared with 64 patients (56%) at 1 year. Fourteen of 30 crossover patients (47%) achieved a good outcome at 1 year and seven (23%) no longer fulfilled criteria for CFS. CONCLUSIONS: Benefits of the intervention were maintained at 2 years. Delaying treatment is associated with reduced efficacy and required more intensive therapy. PMID- 14754827 TI - Prevalence of DSM-IV psychiatric disorder in the French elderly population. AB - BACKGROUND: France has high rates of psychotropic drug consumption and suicide in the elderly population, but it has not yet been possible to determine whether this is due to exceptionally high morbidity rates. AIMS: To describe the first longitudinal population study of psychiatric disorder undertaken in France, and to estimate current and lifetime prevalences and age of onset of psychiatric disorder. METHOD: A study group of 1873 non-institutionalised persons aged 65 years and over was randomly recruited from the Montpellier district electoral rolls. The Mini International Neuropsychiatric Interview was used to assess current and lifetime symptoms. Cases identified by the application of DSM-IV criteria were re-examined by a clinical panel. RESULTS: Forty-six per cent of the study population had experienced a mental disorder in their lifetime, and 3.7% had made a suicide attempt. Lifetime prevalence of major depression was 26.5% and 30% for anxiety disorders. Current prevalence rates were 14.2% for anxiety disorders, 10.7% for phobia, 3% for major depression and 1.7% for psychosis. CONCLUSIONS: Results show very high rates of lifetime but not current major depression. Rates of current phobia and suicidal ideation in the very elderly are also high compared with other studies. The rates reported are likely to be underestimates. PMID- 14754828 TI - Severity of depressive episodes according to ICD-10: prediction of risk of relapse and suicide. AB - BACKGROUND: The ICD-10 categorisation of severity of depression into mild, moderate and severe depressive episodes has not been validated. AIMS: To validate the ICD-10 categorisation of severity of depression by estimating its predictive ability on the course of illness and suicidal outcome. METHOD: All psychiatric in patients in Denmark who had received a diagnosis of a single depressive episode at their first discharge between 1994 and 1999 were identified. The risk of relapse and the risk of suicide were compared for patients discharged with an ICD 10 diagnosis of a single mild, moderate or severe depressive episode. RESULTS: At their first discharge, 1103 patients had an ICD-10 diagnosis of mild depressive episode, 3182 had a diagnosis of moderate depressive episode and 2914 had a diagnosis of severe depressive episode. The risk of relapse and the risk of suicide were significantly different for the three types of depression- increasing from mild to moderate to severe depressive episode. CONCLUSIONS: The ICD-10 way of grading severity is clinically useful and should be preserved in future versions. PMID- 14754829 TI - Antidepressants and public health in Iceland. Time series analysis of national data. AB - BACKGROUND: Major depressive disorder is the second leading cause of disability adjusted life-years in developed regions of the world and antidepressants are the third-ranking therapy class worldwide. AIMS: To test the public health impact of the escalating sales of antidepressants. METHOD: Nationwide data from Iceland are used as an example to study the effect of sales of antidepressants on suicide, disability, hospital admissions and out-patient visits. RESULTS: Sales of antidepressants increased from 8.4 daily defined doses per 1000 inhabitants per day in 1975 to 72.7 in 2000, which is a user prevalence of 8.7% for the adult population. Suicide rates fluctuated during 1950-2000 but did not show any definite trend. Rates for out-patient visits increased slightly over the period 1989-2000 and admission rates increased even more. The prevalence of disability due to depressive and anxiety disorders has not decreased over the past 25 years. CONCLUSIONS: The dramatic increase in the sales of antidepressants has not had any marked impact on the selected public health measures. Obviously, better treatment for depressive disorders is still needed in order to reduce the burden caused by them. PMID- 14754830 TI - Epidemiology of involuntary placement of mentally ill people across the European Union. AB - BACKGROUND: Despite the tendency for harmonisation of strategies for mental health care delivery, rules and regulations for involuntary placement or treatment of mentally ill persons still differ remarkably internationally. Rapid European integration and other political developments require valid and reliable international overviews, sound studies and profound analyses of this controversial issue. AIMS: To give an overview of compulsory admission data from official sources across the European Union (EU). METHOD: Data on the legal frameworks for involuntary placement or treatment of people with mental illness and their outcomes were provided and assessed by experts from all EU member states. RESULTS: Total frequencies of admission and compulsory admission rates vary remarkably across the EU. Variation hints at the influence of differences in legal frameworks or procedures. Time series suggest an overall tendency towards more or less stable quotas in most member states. CONCLUSIONS: Further research is greatly needed in this field. Common international health reporting standards are essential to the compilation of basic data. PMID- 14754831 TI - Trauma exposure and post-traumatic stress symptoms in urban African schools. Survey in CapeTown and Nairobi. AB - BACKGROUND: There is a lack of comparative data on the prevalence and effects of exposure to violence in African youth. AIMS: We assessed trauma exposure, post traumatic stress symptoms and gender differences in adolescents from two African countries. METHOD: A sample of 2041 boys and girls from 18 schools in CapeTown and Nairobi completed anonymous self-report questionnaires. RESULTS: More than 80% reported exposure to severe trauma, either as victims or witnesses. Kenyan adolescents, compared with South African, had significantly higher rates of exposure to witnessing violence (69% v. 58%), physical assault by a family member (27% v. 14%) and sexual assault (18% v. 14%). But rates of current full-symptom post-traumatic stress disorder (PTSD) (22.2% v. 5%) and current partial-symptom PTSD (12% v. 8%) were significantly higher in the South African sample. Boys were as likely as girls to meet PTSD symptom criteria. CONCLUSIONS: Although the lifetime exposure to trauma was comparable across both settings, Kenyan adolescents had much lower rates of PTSD. This difference may be attributable to cultural and other trauma-related variables. High rates of sexual assault and PTSD, traditionally documented in girls, may also occur in boys and warrant further study. PMID- 14754832 TI - Stigma: the feelings and experiences of 46 people with mental illness. Qualitative study. AB - BACKGROUND: Stigma defines people in terms of some distinguishing characteristic and devalues them as a consequence. AIMS: To describe the relationship of stigma with mental illness, psychiatric diagnosis, treatment and its consequences of stigma for the individual. METHOD: Narrative interviews were conducted by trained users of the local mental health services; 46 patients were recruited from community and day mental health services in North London. RESULTS: Stigma was a pervasive concern to almost all participants. People with psychosis or drug dependence were most likely to report feelings and experiences of stigma and were most affected by them. Those with depression, anxiety and personality disorders were more affected by patronising attitudes and feelings of stigma even if they had not experienced any overt discrimination. However, experiences were not universally negative. CONCLUSIONS: Stigma may influence how a psychiatric diagnosis is accepted, whether treatment will be adhered to and how people with mental illness function in the world. However, perceptions of mental illness and diagnoses can be helpful and non-stigmatising for some patients. PMID- 14754833 TI - Drug misuse in pregnancy. PMID- 14754834 TI - Evidence needed for treatment strategies in refractory depression. PMID- 14754835 TI - Personality in psychiatry: what thin partitions? PMID- 14754836 TI - Creativity and mental health. PMID- 14754837 TI - Flashbacks in war veterans. PMID- 14754838 TI - Mental health and social capitals. PMID- 14754839 TI - Creativity, mental disorder and jazz. PMID- 14754842 TI - Evidence, methods, and psychiatric education. PMID- 14754843 TI - Where is psychiatry going and who is going there? PMID- 14754844 TI - Bullish on psychiatry. PMID- 14754845 TI - Psychiatric workforce: past legacies, current dilemmas, and future prospects. PMID- 14754846 TI - Implications of a needs-based approach to estimating psychiatric workforce requirements. AB - The author reviews a needs-based approach to estimating psychiatric workforce requirements that entails five determinations: (1) number of people with mental health problems, (2) number of people needing mental health treatment, (3) number of people needing psychiatric treatment, (4) amount of psychiatric time required to meet patient needs, and (5) amount of time psychiatrists have available to provide direct patient care. Questions, issues, and strategies raised by the needs-based approach are outlined. The author suggests that only a coordinated, carefully orchestrated effort among national psychiatric organizations will ensure that the future psychiatric workforce is adequate to meet the needs of the mentally ill. PMID- 14754847 TI - Selected characteristics and data of psychiatrists in the United States, 2001 2002. AB - OBJECTIVE: To provide basic data about the physician workforce as a whole and the relative place of psychiatrists in the total workforce. To provide data on characteristics of psychiatrists' work activities in routine psychiatric practice. METHOD: Data were obtained from the American Medical Association's (AMA) Physician Characteristics and Distribution in the United States, 2002-2003 and the 2002 National Survey of Psychiatric Practice, a nationally representative survey of 2,000 randomly selected psychiatrists in the United States. RESULTS: Psychiatry is the fourth largest specialty in the United States. Since 1970, psychiatry has grown 86.7%, while child psychiatry has grown 194.6%. However, psychiatrists are distributed unequally across the country, are working fewer hours than in the past, and less of their time is spent in direct patient care activities. CONCLUSIONS: Psychiatry is a growing and significant part of the U.S. physician workforce. However, if trends that show the psychiatric workforce is aging and working fewer hours continue, it is unclear if its current rate of growth will be able to keep pace with the demand for psychiatric services. PMID- 14754848 TI - Recruitment of U.S. medical graduates into psychiatry: reasons for optimism, sources of concern. AB - The authors explain the importance of recruiting U.S. medical graduates into psychiatry, give reasons for optimism about future recruitment, express concerns about problems that could hinder it, and recommend ways to address these concerns. Reasons for optimism include: features of the specialty, such as its focus on the doctor/patient relationship; its increasing job availability and incomes; its scientific achievements; the peaking and possible fading of the National Generalist Initiative; and a 5-year upward recruitment trend. Concerns are: low "overt" interest in psychiatry among entering medical students; clerkship directors' perceptions of a negative educational impact of managed care; graduating seniors' suboptimal satisfaction with their psychiatry clerkships; and what is likely to be a small impact of New Mexico legislation on prescribing privileges for psychologists. The authors make recommendations for addressing these concerns. PMID- 14754849 TI - Factors affecting medical student career choice of psychiatry from 1999 to 2001. AB - OBJECTIVE: The proportion of students matching into psychiatry (PMP) at each medical school results from a complex interplay between extrinsic (e.g., national trends, geographic region) and intrinsic factors (e.g., the quality of psychiatric education). The goal of the study was to learn the extent to which regional and local extrinsic factors (and one intrinsic factor) influenced PMP at medical schools in the U.S. from 1999 to 2001. METHODS: The authors obtained data about these factors from deans of student affairs, the National Resident Matching Program (NRMP), American Medical Association (AMA), Association of American Medical Colleges (AAMC), American Psychiatric Association (APA), and Harvard University's HealthSystem Consortium. RESULTS: The best predictor of a school's PMP is the PMP of the prior year for that particular school. Local and regional extrinsic factors were not significantly associated with PMP. There was a modest inverse correlation between PMP and the proportion of international medical graduates (IMGs) in psychiatry residency. CONCLUSIONS: The authors infer that intrinsic factors are most important for recruitment, and they make recommendations for addressing these factors. PMID- 14754850 TI - Recent trends in psychiatry residency workforce with special reference to international medical graduates. AB - OBJECTIVE: This study examines trends in the supply, distribution, and demographics of psychiatry residents during the 1990s. It evaluates the extent to which the predicted downsizing of psychiatry residency training programs actually occurred and how it affected training programs of different sizes and locations. METHOD: Data for this study were obtained from the American Medical Association's (AMA) Annual Survey of Graduate Medical Education (GME) Programs, the AMA GME directory, and the APA Graduate Medical Census. The study compares the roles played by international medical graduates (IMGs) in contrast to U.S. medical graduates (USMGs) in these trends. RESULTS: There was a significant decline in the number of residents during the years studied. The median training program size also decreased. International medical graduates found broad acceptance in training programs of all locations and sizes, including medical school based programs. Implications of the findings are discussed regarding the impact of current graduate medical education (GME) and immigration policies on future workforce patterns. CONCLUSION: The field will have to decide whether it can afford anymore residency downsizing in light of emerging evidence of a shortage of psychiatrists. PMID- 14754851 TI - Child and adolescent psychiatry workforce: a critical shortage and national challenge. AB - Despite the decades-long projection of an increasing utilization of child and adolescent psychiatry services and an undersupply of child and adolescent psychiatrists, the actual growth and supply of child and adolescent psychiatrists have been very slow. Inadequate support in academic institutions, decreasing graduate medical education (GME) funding, decreasing clinical revenues in the managed care environment, and a devalued image of the profession have made academic child and adolescent psychiatry programs struggle for recruitment of both residents and faculty, although child and adolescent psychiatry has made impressive progress in its scientific knowledge base through research, especially in neuroscience and developmental science. While millions of young people suffer from severe mental illnesses, there are only about 6,300 child and adolescent psychiatrists practicing in the United States. There is also a severe maldistribution of child and adolescent psychiatrists, especially in rural and poor, urban areas where access is significantly reduced. By any method of workforce analysis, it is evident that there will continue to be a shortage of child and adolescent psychiatrists well into the future. Medical/psychiatric educators have a mission to encourage medical students and general psychiatry residents to enter child and adolescent psychiatry and provide crucial mental health care and health care advocacy for our country's youngest and most vulnerable citizens. This article stems from the work of the American Academy of Child and Adolescent Psychiatry Task Force on Work Force Needs, which led to its 10-year recruitment initiative. PMID- 14754852 TI - Teaching normal development using stimulus videotapes in psychiatric education. AB - This article examines the educational principles supporting the use of stimulus videotapes. General principles of using videotape in teaching are discussed, followed by a specific review of the literature regarding the uses of videotape in psychiatric education. The literature on the use of stimulus videotape in psychiatric education is reviewed, with particular attention to its use in teaching normal development. PMID- 14754853 TI - Football is the most popular sport worldwide. PMID- 14754854 TI - Risk factors for injuries in football. AB - BACKGROUND: The injury risk in football is high, but little is known about causes of injury. PURPOSE: To identify risk factors for football injuries using a multivariate model. STUDY DESIGN: Prospective cohort study. METHODS: Participants were 306 male football players from the two highest divisions in Iceland. Before the 1999 football season started, the following factors were examined: height, weight, body composition, flexibility, leg extension power, jump height, peak O(2) uptake, joint stability, and history of previous injury. Injuries and player exposure were recorded throughout the competitive season. RESULTS: Older players were at higher risk of injury in general (odds ratio [OR] = 1.1 per year, P = 0.05). For hamstring strains, the significant risk factors were age (OR = 1.4 [1 year], P < 0.001) and previous hamstring strains (OR = 11.6, P <0.001). For groin strains, the predictor risk factors were previous groin strains (OR = 7.3, P = 0.001) and decreased range of motion in hip abduction (OR = 0.9 [1 degrees ], P = 0.05). Previous injury was also identified as a risk factor for knee (OR = 4.6) and ankle sprains (OR = 5.3). CONCLUSIONS: Age and previous injury were identified as the main risk factors for injury among elite football players from Iceland. PMID- 14754855 TI - An assessment of football referees' decisions in incidents leading to player injuries. AB - PURPOSE: To assess whether match injuries to footballers occurred as a result of players' noncompliance with the rules of the game, match referees could reliably identify the legality of incidents leading to injury, and the rules of football were adequate to protect players from injury. METHODS: Video recordings of incidents leading to injury in 12 FIFA tournaments were used to identify parameters. Team physicians reported the details of match injuries. Two panels of FIFA referees reassessed the legality of incidents from these tournaments that resulted in injuries. RESULTS: In total, 148 general injuries and 84 head/neck injuries were assessed. For the general injuries, the match referees identified 47% and the referees' panel identified 69% as fouls. For head injuries, the match referees identified 40% and the referees' panel identified 49% as fouls. CONCLUSIONS: Decisions made on the legality of tackles leading to injury indicated that the current rules of football were adequate for the majority of tackle situations, although the reliability with which referees could identify fouls during some match conditions was low. For incidents leading to head/neck injuries, the match referees and the referees' panel both identified a smaller proportion of injury situations as fouls. PMID- 14754856 TI - Football injuries during the World Cup 2002. AB - BACKGROUND: The Federation Internationale de Football Association (FIFA) World Cup is one of the largest, most popular sporting events but is associated with a certain risk of injury for the players. PURPOSE: Analysis of the incidence, circumstances, and characteristics of soccer injury during the World Cup 2002. STUDY DESIGN: Prospective survey. METHODS: The physicians of all participating teams reported all injuries after each match on a standardized injury report form. The response rate was 100%. RESULTS: A total of 171 injuries were reported from the 64 matches, which is equivalent to an incidence of 2.7 injuries per match; approximately 1 to 2 injuries per match resulted in absence from training or match. More than a quarter of all injuries were incurred without contact with another player, and 73% were contact injuries. Half of the contact injuries, or 37% of all injuries, were caused by foul play as rated by the team physician and the injured player. CONCLUSION: The incidence of injuries during the World Cup 2002 was similar to those reported for the World Cup in 1994 and in 1998. Increased awareness of the importance of fair play may assist in the prevention of injury. PMID- 14754857 TI - An assessment of player error as an injury causation factor in international football. AB - OBJECTIVE: To quantify the frequency of injury in football as a function of tackle parameters. METHOD: Video recordings of 123 international matches in three Federation Internationale de Football Association tournaments were used to analyze tackle parameters, and team physicians provided reports of postmatch medical attention to players. RESULTS: A total of 8572 tackles were assessed, of which 3464 (40.4%) were fouls. There were 299 incidents of on-pitch medical attention, of which 131 (44%) resulted from foul tackles, and 200 postmatch team physicians' reports, of which 96 (48%) resulted from foul tackles. The cases of on-pitch medical attention resulted in 76 (25%) postmatch physicians' reports, but 124 (62%) of the postmatch physicians' reports were not associated with on pitch medical attention. Tackled players received 74% of the postmatch medical reports. Tackle types with the greatest probability of requiring medical attention were from the side in terms of tackle direction, jumping vertically in terms of tackle mode, and a clash of heads in terms of tackle action. CONCLUSION: Human error on the part of players during the process of tackling and inadequacies in the laws of football and/or their application by match referees were equally responsible for the high levels of injury observed. PMID- 14754858 TI - Football injuries at Asian tournaments. AB - PURPOSE: To examine the incidences and patterns of injuries that required medical attention among Asian football players. METHODS: A total of 411 Asian football players at both senior and youth (U-20) elite levels were observed during 50 international matches. Independent injury observers and team doctors determined the occurrence of injuries and recorded the location, type, time, and circumstances of the injuries using a protocol sheet. RESULTS: The overall injury frequency rate was 45.8 out of 1000 hours. As the tournaments progressed into the knockout stages, the incidence and severity of the injuries increased. The most common sites of injuries were the knees (18.5%), lower legs (17.3%), and ankles (14.2%). Although most injuries were diagnosed as contusions, the more serious injuries were those diagnosed as sprains (especially concerning the knee and ankle) or strains (thigh and back). CONCLUSIONS: The incidences of injuries to Asian football players were higher than those to European players, but the patterns of the injuries showed no major differences. CLINICAL RELEVANCE: To develop an injury-prevention program, more solid and comprehensive data need to be collected to identify the risk factors and variables associated with higher incidences of injuries to Asian football players. PMID- 14754859 TI - The influence of tackle parameters on the propensity for injury in international football. AB - PURPOSE: To understand how tackling leads to injury in football, to develop a framework for classifying tackles, and to identify tackles with the greatest propensity to cause injury. METHOD: Video recordings of 123 matches in three FIFA tournaments were used to identify tackling parameters. Team physicians prepared reports of postmatch medical attention to players. RESULTS: Tackles from the side were twice as likely to require postmatch medical attention as tackles from behind. Injuries to the head/neck of tackled and tackling players and the torso of tackling players were more likely to receive on-pitch medical attention than other injuries. Injuries to the foot for tackled and tackling players and the lower leg and thigh for tackling players were less likely to receive on-pitch medical attention than other injuries. Tackles with the greatest propensity for causing injury involved clash of heads and two-footed tackles for tackled players and clash of heads, two-footed tackles, jumping vertically, and tackles from the side for tackling players. CONCLUSION: The laws of football relating to tackling should be reviewed to provide greater protection from injury by reducing the overall level of risk and, in particular, by protecting players from tackles with the highest propensity for causing injury. PMID- 14754860 TI - The impact of specific high-intensity training sessions on football referees' fitness levels. AB - BACKGROUND: In comparison to the amount of literature that has examined the match demands of football refereeing, there has been little attempt to assess the impact of high-intensity training. PURPOSE: The main goals were to get a better understanding of the long-term effect of specific intermittent training. STUDY DESIGN: The authors examined the cardiovascular strain of specific high-intensity training sessions and also their impact on referees' fitness levels. METHODS: To examine the physical workload during intensive intermittent training sessions, heart rates were recorded and analyzed relative to the referees' maximum heart rate (HR(max)). To assess the referees' fitness levels, the Yo-Yo intermittent recovery test was used. RESULTS: Both the pitch- and track-training sessions were successful in imposing an appropriate high intensity load on the referees, at 86.4 +/- 2.9% and 88.2 +/- 2.4% HR(max), respectively. Following 16 months of intermittent high-intensity training, referees improved their performance on the Yo-Yo intermittent recovery test by 46.5%, to a level that is comparable with professional players. CONCLUSIONS: As match officials are subjected to a high physical load during matches, they should follow structured weekly training plans that have an emphasis on intensive, intermittent training sessions. PMID- 14754861 TI - Rule violations as a cause of injuries in male norwegian professional football: are the referees doing their job? AB - BACKGROUND: Foul play is an important cause of injury in football. Reduction of foul play and adherence to the laws of the game may be possible interventions to reduce the rate of injuries. PURPOSE: To evaluate how violations of the laws of the game contribute to injury and to investigate whether the decisions made by the referees are correct in high-risk situations. STUDY DESIGN: Prospective cohort study. METHODS: Videotapes and injury information were collected for 174 of 182 matches from the male Norwegian professional league during the 2000 season. Three Norwegian FIFA referees performed retrospective blinded evaluation of the 406 incidents. RESULTS: Less than one-third of the injuries identified on video and about 40% of the incidents with a high risk of injury resulted in a free kick being awarded. About 1 in 10 of these situations led to either a yellow or red card. The agreement between decisions made by the match referee and the expert referee panel was good, that is, their decisions agreed in 85% of the situations in which injury occurred. CONCLUSIONS: There may be a need for an improvement of the laws of the game of football to protect players from dangerous play. PMID- 14754862 TI - Video analysis of the mechanisms for ankle injuries in football. AB - BACKGROUND: Although ankle sprains are frequent in football, little is known about their mechanisms. PURPOSE: To describe the injury mechanisms for ankle injuries in male elite football. STUDY DESIGN: Prospective cohort study. METHODS: Videotapes and injury information were collected for 313 of 409 matches from Norwegian and Icelandic elite football during the 1999 to 2000 seasons. Video recordings of incidents that resulted in ankle injuries were analyzed and cross referenced with injury reports from the team medical staff. RESULTS: A total 46 acute ankle injuries were reported to have occurred, that is, 4.5 injuries per 1000 match hours. Of these, 26 (57%) were identified on the videotapes. Two mechanisms thought to be specific to football were found: 1) player-to-player contact with impact by an opponent on the medial aspect of the leg just before or at foot strike, resulting in a laterally directed force causing the player to land with the ankle in a vulnerable, inverted position; and 2) forced plantar flexion where the injured player hit the opponent's foot when attempting to shoot or clear the ball. CONCLUSIONS: Systematic video analysis provides detailed information on the mechanisms for ankle injuries in football-for lateral ligament sprains and for the condition dubbed "footballer's ankle." PMID- 14754863 TI - Football injuries during FIFA tournaments and the Olympic Games, 1998-2001: development and implementation of an injury-reporting system. AB - BACKGROUND: Standardized assessment of sports injuries provides not only important epidemiological information, but also directions for injury prevention, and the opportunity for monitoring long-term changes in the frequency and characteristics of injury. PURPOSE: Development and implementation of an easy to use injury-reporting system to analyze the incidence, circumstances and characteristics of injury during major international football tournaments. STUDY DESIGN: Prospective survey. METHODS: A comprehensive injury report form was developed, and implemented during 12 international football tournaments. The physicians of all participating teams were asked to report all injuries after each match. The response rate was 84% on average. RESULTS: A total of 901 injuries were reported from 334 matches, which is equivalent to an incidence of 2.7 injuries per match. Approximately one injury per match resulted in a player's absence from training or matches. On average 86% of the injuries arose as a result of contact with another player, and approximately half of all injuries were caused by foul play. The number of injuries per match differed substantially between the tournaments for players of different age, sex and skill-level. CONCLUSION: An injury-reporting system has been implemented as matter of routine in FIFA tournaments. The consistent findings in the present study demonstrate the high quality of the data obtained. PMID- 14754864 TI - The parahippocampal gyrus in the baboon: anatomical, cytoarchitectonic and magnetic resonance imaging (MRI) studies. AB - The parahippocampal gyrus, located at the medial temporal lobe, is a key structure in declarative memory processing. We have analyzed the general organization of the parahippocampal gyrus in the baboon, a nonhuman primate species relatively close to human. This region is rostrocaudally made up of the temporopolar, perirhinal, entorhinal (divided into seven subfields) and posterior parahippocampal (areas TH and TF) cortices. The basic analysis has been performed in three brains, serially sectioned and stained with thionin, myelin stain, acetylcholinesterase and parvalbumin, to determine cytoarchitectonic boundaries. Borders of all subfields were charted onto camera lucida drawings, and two dimensional maps of the surface and topography of the parahippocampal gyrus were made. Finally, the limits of each parahippocampal area were then transposed on corresponding MR images (commonly used for in vivo PET or functional MRI activation studies) of two animals for precise identification. The general cytoarchitectonic features of the baboon parahippocampal gyrus are similar to macaques, but the size of temporopolar cortex and the laminar organization of perirhinal and posterior parahippocampal cortices resemble humans more than macaque species. In conclusion, the size and structure of the baboon parahippocampal cortex makes this species very appropriate for experimental studies on memory function. PMID- 14754865 TI - Contributions of sensory input, auditory search and verbal comprehension to cortical activity during speech processing. AB - We studied eight normal subjects in an fMRI experiment where they listened to natural speech sentences and to matched simple or complex speech envelope noises. Neither of the noises (simple or complex) were understood initially, but after the corresponding natural speech sentences had been heard, comprehension was close to perfect for the complex but still absent for the simple speech envelope noises. This setting thus involved identical stimuli that were understood or not and permitted to identify (i) a neural substrate of speech comprehension unconfounded by stimulus acoustic properties (common to natural speech and complex noises), (ii) putative correlates of auditory search for phonetic cues in noisy stimuli (common to simple and complex noises once the matching natural speech had been heard) and (iii) the cortical regions where speech comprehension and auditory search interact. We found correlates of speech comprehension in bilateral medial (BA21) and inferior (BA38 and BA38/21) temporal regions, whereas acoustic feature processing occurred in more dorsal temporal regions. The left posterior superior temporal cortex (Wernicke's area) responded to the acoustic complexity of the stimuli but was additionally sensitive to auditory search and speech comprehension. Attention was associated with recruitment of the dorsal part of Broca's area (BA44) and interaction of auditory attention and comprehension occurred in bilateral insulae, the anterior cingulate and the right medial frontal cortex. In combination, these results delineate a neuroanatomical framework for the functional components at work during natural speech processing, i.e. when comprehension results from concurrent acoustic processing and effortful auditory search. PMID- 14754866 TI - Temporal and cerebellar brain regions that support both declarative memory formation and retrieval. AB - Using event-related fMRI, we scanned young healthy subjects while they memorized real-world photographs and subsequently tried to recognize them within a series of new photographs. We confirmed that activity in the medial temporal lobe (MTL) and inferior prefrontal cortex correlates with declarative memory formation as defined by the subsequent memory effect, stronger responses to subsequently remembered than forgotten items. Additionally, we confirmed that activity in specific regions within the parietal lobe, anterior prefrontal cortex, anterior cingulate and cerebellum correlate with recognition memory as measured by the conventional old/new effect, stronger responses for recognized old items (hits) than correctly identified new items (correct rejections). To obtain a purer measure of recognition success, we introduced two recognition effects by comparing brain responses to hits and old items misclassified as new (misses). The positive recognition effect (hits > misses) revealed prefrontal, parietal and cerebellar contributions to recognition, and in line with electrophysiological findings, the negative recognition effect (hits < misses) revealed an anterior medial temporal contribution. Finally, by inclusive masking, we identified temporal and cerebellar brain areas that support both declarative memory formation and retrieval. For matching operations during recognition, these areas may re-use representations formed and stored locally during encoding. PMID- 14754867 TI - The distribution of immunoreactivity for intracellular androgen receptors in the cerebral cortex of hormonally intact adult male and female rats: localization in pyramidal neurons making corticocortical connections. AB - Gonadal hormones are known to broadly influence cortical information processing. Findings from this study in rats suggest that for androgens, this influence may include stimulation of underlying corticocortical connections. First, immunoreactivity for intracellular androgen receptors, while present in all regions and layers examined, was found to be particularly abundant in sensory and motor regions, and within these, within their major pyramidal cell layers, i.e. layers II/III and V/VI. Double labeling immunocytochemical studies for androgen receptors and for neuron-specific markers then confirmed that the majority of receptor-bearing cortical cells were pyramidal neurons. Finally, combined analyses of cortical receptor immunoreactivity and retrograde labeling produced by tracer injections made in specific subcortical (caudate, nucleus accumbens, superior colliculus, thalamus) areas yielded only isolated examples of receptor/tracer overlap. However, injections made within the cortex itself (sensory, motor, associational areas) retrogradely labeled cortical cells some 50% or more - especially within injected hemispheres, were receptor immunoreactive. Thus, the regional, laminar, and cellular distributions of immunoreactivity in the rat cerebrum largely identify pyramidal neurons with connectional signatures aligning intracellular androgen receptors with the local, associational, and to a lesser degree, callosal circuits that interlink territories of the cortical mantle and play key roles in cortical information processing. PMID- 14754868 TI - Co-expression and in vivo interaction of serotonin1A and serotonin2A receptors in pyramidal neurons of prefrontal cortex. AB - The prefrontal cortex plays a key role in the control of higher brain functions and is involved in the pathophysiology and treatment of schizophrenia. Here we report that approximately 60% of the neurons in rat and mouse prefrontal cortex express 5-HT(1A) and/or 5-HT2A receptor mRNAs, which are highly co-localized (approximately 80%). The electrical stimulation of the dorsal and median raphe nuclei elicited 5-HT1A-mediated inhibitions and 5-HT2A-mediated excitations in identified pyramidal neurons recorded extracellularly in rat medial prefrontal cortex (mPFC). Opposite responses in the same pyramidal neuron could be evoked by stimulating the raphe nuclei at different coordinates, suggesting a precise connectivity between 5-HT neuronal subgroups and 5-HT1A and 5-HT2A receptors in pyramidal neurons. Microdialysis experiments showed that the increase in local 5 HT release evoked by the activation of 5-HT2A receptors in mPFC by DOI (5-HT2A/2C receptor agonist) was reversed by co-perfusion of 5-HT1A agonists. This inhibitory effect was antagonized by WAY-100635 and the prior inactivation of 5 HT1A receptors in rats and was absent in mice lacking 5-HT1A receptors. These observations help to clarify the interactions between the mPFC and the raphe nuclei, two key areas in psychiatric illnesses and improve our understanding of the action of atypical antipsychotics, acting through these 5-HT receptors. PMID- 14754869 TI - Perception-related modulations of local field potential power and coherence in primary visual cortex of awake monkey during binocular rivalry. AB - Cortical synchronization at gamma-frequencies (35-90 Hz) has been proposed to define the connectedness among the local parts of a perceived visual object. This hypothesis is still under debate. We tested it under conditions of binocular rivalry (BR), where a monkey perceived alternations among conflicting gratings presented singly to each eye at orthogonal orientations. We made multi-channel microelectrode recordings of multi-unit activity (MUA) and local field potentials (LFP) from striate cortex (V1) during BR while the monkey indicated his perception by pushing a lever. We analyzed spectral power and coherence of MUA and LFP over 4-90 Hz. As in previous work, coherence of gamma-signals in most pairs of recording locations strongly depended on grating orientation when stimuli were presented congruently in both eyes. With incongruent (rivalrous) stimulation LFP power was often consistently modulated in consonance with the perceptual state. This was not visible in MUA. These perception-related modulations of LFP occurred at low and medium frequencies (< 30 Hz), but not at gamma-frequencies. Perception-related modulations of LFP coherence were also restricted to the low-medium range. In conclusion, our results do not support the expectation that gamma-synchronization in V1 is related to the perceptual state during BR, but instead suggest a perception-related role of synchrony at low and medium frequencies. PMID- 14754870 TI - Neural responses during interception of real and apparent circularly moving stimuli in motor cortex and area 7a. AB - We recorded the neuronal activity in the arm area of the motor cortex and parietal area 7a of two monkeys during interception of stimuli moving in real and apparent motion. The stimulus moved along a circular path with one of five speeds (180-540 degrees/s), and was intercepted at 6 o'clock by exerting a force pulse on a semi-isometric joystick which controlled a cursor on the screen. The real stimuli were shown in adjacent positions every 16 ms, whereas in the apparent motion situation five stimuli were flashed successively at the vertices of a regular pentagon. The results showed, first, that a group of neurons in both areas above responded not only during the interception but also during a NOGO task in which the same stimuli were presented in the absence of a motor response. This finding suggests these areas are involved in both the processing of the stimulus as well as in the preparation and production of the interception movement. In addition, a group of motor cortical cells responded during the interception task but not during a center --> out task, in which the monkeys produced similar force pulses towards eight stationary targets. This group of cells may be engaged in sensorimotor transformations more specific to the interception of real and apparent moving stimuli. Finally, a multiple regression analysis revealed that the time-varying neuronal activity in area 7a and motor cortex was related to various aspects of stimulus motion and hand force in both the real and apparent motion conditions, with stimulus-related activity prevailing in area 7a and hand-related activity prevailing in motor cortex. In addition, the neural activity was selectively associated with the stimulus angle during real motion, whereas it was tightly correlated to the time-to-contact in the apparent motion condition, particularly in the motor cortex. Overall, these observations indicate that neurons in motor cortex and area 7a are processing different parameters of the stimulus depending on the kind of stimulus motion, and that this information is used in a predictive fashion in motor cortex to trigger the interception movement. PMID- 14754871 TI - Recovery of evoked potentials, metabolic activity and behavior in a mouse model of somatosensory cortex lesion: role of the neural cell adhesion molecule (NCAM). AB - Understanding the processes that underlie functional recovery after cortical injury is a major challenge for neurobiology and clinical neurology. The aim of the present study was to establish a mouse model of functional recovery that would facilitate the investigation of the molecular and cellular events involved in cortical dynamics. We show that a focal injury of approximately 0.5 mm of diameter and 1 mm depth made in the barrel cortex of adult mice induced a transitory deficit that could be characterized using somatosensory evoked potential (SEP), metabolic mapping and a behavioral test. SEP recordings of short latency responses using an epicranial multi-array system showed a decreased cortical activity in the peri-lesion regions 2 weeks after the injury and a partial recovery to normal pattern 6 weeks after the lesion. Delayed SEP signals over the motor cortex were not altered by the injury. Metabolic mapping with [14C]deoxyglucose uptake in the surround of the injury reproduced the time course of deficit and recovery. Finally, a deficit in vibrissae related performance in a gap-crossing test 1 week after injury was followed by a functional recovery in the following 2 weeks. We show in addition that the recovery process is deficient and significantly delayed in NCAM knockout mice lacking all isoforms of NCAM (neural cell adhesion molecule)and PSA-NCAM. These results support the hypothesis that impairment and recovery of functions after focal cortical lesion involves remodeling of intact circuits surrounding the lesion and that the NCAM molecule participate in this process. The model opens new possibilities for investigating the role of candidate molecules in functional recovery using genetically modified mice. PMID- 14754872 TI - Parvalbumin expression in visual cortical interneurons depends on neuronal activity and TrkB ligands during an Early period of postnatal development. AB - The differentiation of cortical interneurons is controlled by environmental factors. Here, we describe the role of activity and neurotrophins in regulating parvalbumin (PARV) expression using organotypic cultures (OTC) of rat visual cortex as model system. In OTC, PARV expression was dramatically delayed. The organotypic proportion of approximately 6% PARV neurons was not established before 50-70 DIV, whereas in vivo all neurons are present until P20. Thalamic afferents increased cortical PARV mRNA in OTC, but not to the age-matched in vivo level. During the first 10 DIV, BDNF and NT-4 accelerated PARV mRNA expression in a Trk receptor and MEK2 dependent manner. The BDNF action required PI3 kinase signalling. PARV expression required activity. The proportion of neurons which managed to up-regulate PARV was inversely related to the duration of early transient periods of activity deprivation. Long-term activity-deprived OTC completely failed to up-regulate PARV mRNA. Both TrkB ligands failed to promote PARV expression in activity-deprived OTC. However, a few basket and chandelier neurons were observed, suggesting that the development of class-specific morphological features is activity-independent. Once established, PARV expression became resistant to late-onset activity deprivation. In conclusion, PARV expression depended on activity and TrkB ligands which appear to prime the PARV expression already before its developmental onset. PMID- 14754873 TI - Thioproline inhibits development of esophageal adenocarcinoma induced by gastroduodenal reflux in rats. AB - Several epidemiological cohort studies have suggested that duodeno gastroesophageal reflux per se induces Barrett's esophagus leading to increased risk of the development of esophageal adenocarcinoma (EAC). However, the exact causative factors behind EAC remain unclear. Recently, we designed a new duodenal contents reflux model which retained normal stomach function. In this model, duodenal contents flowed back into the esophagus and stomach resulting in repeated re-entry into the esophagus through the site of esophagojejunostomy. To elucidate the factors underlying the development of EAC, thiazolidine-4 carboxylic acid (thioproline, TPRO) was applied to the new reflux models as a nitrite scavenger and as a probe to detect reactive nitrogen species (RNS). Post operatively, 31 animals were divided into two groups according to diet. Animals belonging to the control group were given normal diet (n = 18), while the TPRO group was given food containing 0.5% TPRO (n = 13). All esophageal sections in both groups were examined using hematoxylin and eosin staining and immunohistochemical analysis of inducible nitric oxide synthase (iNOS). EACs developed in 7 of 18 rats (38.9%) of the control group, whereas no EACs were detected in the TPRO group (Fisher's exact test, P < 0.05). Conversely, esophageal squamous cell carcinoma (ESCC) was detected in 1 of 18 rats (5.6%) of the control group and in 1 of 13 rats (7.7%) of the TPRO group. The incidence of ESCC was not significantly different between the two groups (P = 0.671). iNOS protein was overexpressed in Barrett's esophagus of both groups. The present results suggest that RNS such as nitric oxide and peroxynitrite and nitroso compounds derived from reflux of duodenal contents play an important role in the development of EAC, and that the primary causes of ESCC and EAC may differ. PMID- 14754874 TI - Use of genetically manipulated Salmonella typhimurium strains to evaluate the role of sulfotransferases and acetyltransferases in nitrofen mutagenicity. AB - Nitrofen had been used as a herbicide, until its carcinogenic and teratogenic activity in rodents was detected. A food contamination occurring in 2002 in Germany led to the initiation of new studies in order to better understand the potential risk for humans. Nitrofen is a nitroarene and as such might be activated to a mutagen via reduction to the corresponding hydroxylamine and subsequent formation of a reactive acetic or sulfuric acid ester. Therefore, we have investigated the mutagenicity of nitrofen in Salmonella typhimurium strains engineered for the expression of all human xenobiotic-metabolizing sulfotransferases (SULTs) and acetyltransferases (NATs) identified. Nitrofen was inactive in the parental strains TA1538, TA98 and TA100, but was mutagenic even at low doses when human sulfotransferase SULT1A1 (the major broad-spectrum phenol SULT) was expressed in these strains, but not when it was expressed in a TA1538 derived strain deficient in an endogenous nitroreductase. Several other human SULTs (in particular 1A3 and 1C1) as well as human NAT2 (unlike NAT1) also activated nitrofen, but were markedly less efficient than SULT1A1. Likewise, expression of rat and mouse SULT1A1 led to weaker mutagenic activity of nitrofen than expression of the corresponding human enzyme. An endogenous acetyltransferase only activated nitrofen to a mutagen when it was strongly over expressed in the TA98-derived strain YG1024. Thus, humans might be more susceptible to the carcinogenic effects of nitrofen than mice and rats, which have been used in long-term studies. The fact that several SULTs show particular high expression in fetal tissues suggests that this activation pathway may also play a role in the teratogenic effects observed. PMID- 14754875 TI - Ascorbic acid suppresses drug-induced apoptosis in human colon cancer cells by scavenging mitochondrial superoxide anions. AB - Although a high alimentary intake of antioxidant vitamins such as ascorbic acid may play an important role in cancer prevention, a high level of antioxidants may have quite different effects at different stages of the transformation process. In cancer development, the resistance of cells to apoptosis is one of the most crucial steps. We have tested the effects of ascorbic acid on apoptosis in HT-29 human colon carcinoma cells when induced by two potent apoptosis inducers, the classical antitumor drug camptothecin or the flavonoid flavone. Apoptosis was assessed based on caspase-3-like activity, plasma membrane disintegration and finally nuclear fragmentation and chromatin condensation. Ascorbic acid dose dependently inhibited the apoptotic response of cells to camptothecin and flavone. RT-PCR analysis and western blot analysis revealed that ascorbic acid specifically blocked the decrease of bcl-X(L) by camptothecin or flavone. An increased generation of mitochondrial O(2)(-.) precedes the down-regulation of bcl-X(L) by camptothecin and flavone and ascorbic acid at a concentration of 1 mM prevented the generation of this reactive oxygen species. In conclusion, ascorbic acid functions as a potent antioxidant in mitochondria of human colon cancer cells and thereby blocks drug-mediated apoptosis induction allowing cancer cells to become insensitive to chemotherapeutics. PMID- 14754876 TI - Melatonin uptake and growth prevention in rat hepatoma 7288CTC in response to dietary melatonin: melatonin receptor-mediated inhibition of tumor linoleic acid metabolism to the growth signaling molecule 13-hydroxyoctadecadienoic acid and the potential role of phytomelatonin. AB - Both physiological and pharmacological levels of the pineal hormone melatonin exhibit substantial anticancer activity in tissue-isolated rat hepatoma 7288CTC via melatonin receptor-mediated blockade of tumor uptake of linoleic acid (LA) and its metabolism to the mitogenic signaling molecule 13-hydroxyoctadecadienoic acid (13-HODE). Melatonin is also present in significant amounts in edible plants and is supplied in nutritional supplements. We confirmed the presence of significant quantities of melatonin in 20 varieties of edible plants. In pinealectomized tumor-free rats, 3 weeks of ingestion of either 5 or 50 microg/day of melatonin contained in a semi-purified diet resulted in a dose dependent elevation in steady-state plasma melatonin levels within the nocturnal physiological range. In pineal-intact tumor-bearing rats, the daily intake of 5 microg/day of melatonin for 3 weeks resulted in an enhanced amplitude and duration of the nocturnal melatonin levels within physiological circulating limits. The nocturnal melatonin amplitude in rats ingesting 500 ng of melatonin/day remained within the physiological range. A dose-related increase in tumor concentrations of melatonin occurred in animals ingesting melatonin from the diet. Perfusion of tumors in situ with physiological, nocturnal blood levels of melatonin resulted in a mean 31% uptake and retention of the melatonin. Chronic ingestion of 50 ng, 500 ng or 5 microg of melatonin/day supplied in a semi-purified 5% corn oil diet led to a significant dose-dependent reduction in the rates of tumor total fatty acid uptake, LA uptake, 13-HODE production and tumor growth. The co-ingestion of melatonin receptor antagonist S20928 completely blocked the effects and prevented the intra-tumoral accumulation of melatonin. Melatonin receptor-mediated suppression of tumor growth, LA uptake and metabolism, and stimulation of tumor melatonin uptake and retention in response to the dietary intake of phytomelatonin from edible plants or melatonin from nutritional supplements, could play an important role in cancer growth prevention. PMID- 14754877 TI - Involvement of V(D)J recombinase in the generation of intragenic deletions in the Rit1/Bcl11b tumor suppressor gene in gamma-ray-induced thymic lymphomas and in normal thymus of the mouse. AB - Mouse thymic lymphomas induced by gamma-irradiation exhibited homozygous deletions of the Rit1/Bcl11b tumor suppressor gene on chromosome 12 at high frequencies. Internal deletions of one allele were frequently accompanied by loss of the other allele. In order to elucidate the mechanism of these internal deletions, the sites of breakage and rejoining were examined by PCR mapping and sequencing. The 5' site of the deletions clustered within an approximately 5 kb region of intron 1 and the 3' site was confined to a site in intron 3. These sites contained P and/or N nucleotides and cryptic sequences recognizable by the RAG1/2 recombinase in the vicinity. This suggests that the Rit1 intragenic deletions were generated by endogenous illegitimate V(D)J recombinase activity and such aberrant recombination was also detected by nested PCR of DNA from the thymus of unirradiated mice but not of RAG2-deficient mice. A rough estimate indicated that there reside as many as 10(3)-10(4) thymocytes having Rit1 deletions, assuming the presence of 10(8) thymocytes in the thymus of unirradiated mice. Moreover, the recombination frequency was not affected by gamma-irradiation. These results show no effect of radiation on Rit1 mutations and suggest an indirect mechanism for its role in lymphomagenesis. PMID- 14754878 TI - COX-2 gene promoter haplotypes and prostate cancer risk. AB - Cyclooxygenase-2 (COX-2) is a key rate-limiting enzyme that converts arachidonic acid into pro-inflammatory prostaglandins. COX-2 expression is strongly correlated with increased tumor microvasculature density and plays an important role in inhibiting apoptosis, stimulating angiogenesis and promoting tumor cell metastasis and invasion. However, little is known about the role that sequence variation of the COX-2 gene contributes to prostate cancer. Thus, we searched for polymorphisms in the promoter region of the COX-2 gene using denaturing high performance liquid chromatography. Four single nucleotide polymorphisms (SNPs), 1285A/G, -1265G/A, -899G/C and -297C/G, were detected and confirmed by direct sequencing. Three of the SNPs in the promoter region of COX-2 gene create at least three putative transcription factor binding sites and eliminate CCAAT/enhancer binding protein alpha (C/EBP alpha) and NF-kappa B binding sites. A case-control study of the four SNPs in African American (n = 288), Bini Nigerian (n = 264) and European American (n = 184) prostate cancer cases and age matched controls revealed that SNP -297G was associated with a decreased risk for prostate cancer [odds ratio (OR) = 0.49; CI = 0.2-0.9; P = 0.01]. The effect on risk was observed in both African Americans (OR = 0.51; CI = 0.2-0.9; P = 0.01) and European Americans (OR = 0.33; CI = 0.1-0.9; P = 0.02). In addition, SNPs 1265A and -899C were associated with increased prostate cancer risk in African Americans (OR = 2.72; CI = 1.3-5.8; P = 0.007 and OR = 3.67; CI = 1.4-9.9; P = 0.007, respectively). Haplotype analyses revealed modest effects on susceptibility to prostate cancer across populations. Haplotype GGCC conferred increased risk in the African American and Nigerian populations. Conversely, haplotype AGGG exhibited a negative association with prostate cancer risk in African Americans (OR = 0.4; CI = 0.1-0.9; P = 0.02) and European Americans (OR = 0.2; CI = 0.1-0.9; P = 0.03). These data suggest that variation of the COX-2 promoter may influence the risk and development of prostate cancer. PMID- 14754879 TI - Transforming growth factor-beta1 inhibition of vascular smooth muscle cell activation is mediated via Smad3. AB - Activation of vascular smooth muscle cells (VSMCs) by proinflammatory cytokines is a key feature of atherosclerotic lesion formation. Transforming growth factor (TGF)-beta1 is a pleiotropic growth factor that can modulate the inflammatory response in diverse cell types including VSMCs. However, the mechanisms by which TGF-beta1 is able to mediate these effects remains incompletely understood. We demonstrate here that the ability of TGF-beta1 to inhibit markers of VSMC activation, inducible nitric-oxide synthase (iNOS) and interleukin (IL)-6, is mediated through its downstream effector Smad3. In reporter gene transfection studies, we found that among a panel of Smads, Smad3 could inhibit iNOS induction in an analogous manner as exogenous TGF-beta1. Adenoviral overexpression of Smad3 potently repressed inducible expression of endogenous iNOS and IL-6. Conversely, TGF-beta1 inhibition of cytokine-mediated induction of iNOS and IL-6 expression was completely blocked in Smad3-deficient VSMCs. Previous studies demonstrate that CCAAT/enhancer-binding protein (C/EBP) and NF-kappaB sites are critical for cytokine induction of both the iNOS and IL-6 promoters. We demonstrate that the inhibitory effect of Smad3 occurs via a novel antagonistic effect of Smad3 on C/EBP DNA-protein binding and activity. Smad3 mediates this effect in part by inhibiting C/EBP-beta and C/EBP-delta through distinct mechanisms. Furthermore, we find that Smad3 prevents the cooperative induction of the iNOS promoter by C/EBP and NF-kappaB. These data demonstrate that Smad3 plays an essential role in mediating TGF-beta1 anti-inflammatory response in VSMCs. PMID- 14754880 TI - Hypoxia inhibits myogenic differentiation through accelerated MyoD degradation. AB - Cells undergo a variety of biological responses when placed in hypoxic conditions, including alterations in metabolic state and growth rate. Here we investigated the effect of hypoxia on the ability of myogenic cells to differentiate in culture. Exposure of myoblasts to hypoxia strongly inhibited multinucleated myotube formation and the expression of differentiation markers. We showed that hypoxia reversibly inhibited MyoD, Myf5, and myogenin expression. One key step in skeletal muscle differentiation involves the up-regulation of the cell cycle-dependent kinase inhibitors p21 and p27 as well as the product of the retinoblastoma gene (pRb). Myoblasts cultured under hypoxic conditions in differentiation medium failed to up-regulate both p21 and pRb despite the G1 cell cycle arrest, as evidenced by p27 accumulation and pRb hypophosphorylation. Hypoxia-dependent inhibition of differentiation was associated with MyoD degradation by the ubiquitin-proteasome pathway. MyoD overexpression in C2C12 myoblasts overrode the differentiation block imposed by hypoxic conditions. Thus, hypoxia by inducing MyoD degradation blocked accumulation of early myogenic differentiation markers such as myogenin and p21 and pRb, preventing both permanent cell cycle withdraw and terminal differentiation. Our study revealed a novel anti-differentiation effect exerted by hypoxia in myogenic cells and identified MyoD degradation as a relevant target of hypoxia. PMID- 14754881 TI - Cardiac expression of the cystic fibrosis transmembrane conductance regulator involves novel exon 1 usage to produce a unique amino-terminal protein. AB - Cystic fibrosis is caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene, which encodes a chloride channel present in many cells. In cardiomyocytes, we report that multiple exon 1 usage and alternative splicing produces four CFTR transcripts, with different 5' untranslated regions, CFTR(TRAD-139), CFTR(-1C/-1A), CFTR(-1C), and CFTR(-1B). CFTR transcripts containing the novel upstream exons (exons -1C, -1B, and -1A) represent more than 90% of cardiac expressed CFTR mRNA. Regulation of cardiac CFTR expression, in response to developmental and pathological stimuli, is exclusively due to the modulation of CFTR(-1C) and CFTR(-1C/-1A) expression. Upstream open reading frames have been identified in the 5'-untranslated regions of all CFTR transcripts that, in conjunction with adjacent stem-loop structures, modulate the efficiency of translation initiation at the AUG codon of the main CFTR coding region in CFTR(TRAD-139) and CFTR(-1C/-1A) transcripts. Exon -1A, only present in CFTR(-1C/-1A) transcripts, encodes an AUG codon that is in-frame with the main CFTR open reading frame, the efficient translation of which produces a novel CFTR protein isoform with a curtailed amino terminus. As the expression of this CFTR transcript parallels the spatial and temporal distribution of the cAMP-activated whole-cell current density in normal and diseased hearts, we suggest that CFTR(-1C/-1A) provides the molecular basis for the cardiac cAMP-activated chloride channel. Our findings provide further insight into the complex nature of in vivo CFTR expression, to which multiple mRNA transcripts, protein isoforms, and post-transcriptional regulatory mechanisms are now added. PMID- 14754882 TI - Physiological and biochemical defects in functional interactions of mitochondrial DNA polymerase and DNA-binding mutants of single-stranded DNA-binding protein. AB - Functional interactions between mitochondrial DNA polymerase (pol gamma) and mitochondrial single-stranded DNA-binding protein (mtSSB) from Drosophila embryos greatly enhance the overall activity of pol gamma by increasing primer recognition and binding and stimulating the rate of initiation of DNA strands (Farr, C. L., Wang, Y., and Kaguni, L. S. (1999) J. Biol. Chem. 274, 14779 14785). We show here that DNA-binding mutants of mtSSB are defective in stimulation of DNA synthesis by pol gamma. RNAi knock-down of mtSSB reduces expression to <5% of its normal level in Schneider cells, resulting in growth defects and in the depletion of mitochondrial DNA (mtDNA). Overexpression of mtSSB restores cell growth rate and the copy number of mtDNA, whereas overexpression of a DNA-binding and functionally impaired form of mtSSB neither rescues the cell growth defect nor the mtDNA depletion phenotype. Further development of Drosophila animal models, in which induced mtDNA depletion is manipulated by controlling exogenous expression of wild-type or mutant forms, will offer new insight into the mechanism and progression of human mtDNA depletion syndromes and possible intervention schemes. PMID- 14754883 TI - Importance of transmembrane segment M1 of the sarcoplasmic reticulum Ca2+-ATPase in Ca2+ occlusion and phosphoenzyme processing. AB - The functional consequences of a series of point mutations in transmembrane segment M1 of sarcoplasmic reticulum Ca2+-ATPase were analyzed in steady-state and transient kinetic experiments examining the partial reaction steps involved in Ca2+ interaction and phosphoenzyme turnover. Arginine or leucine substitution of Glu51, Glu55, or Glu58, located in the N-terminal third of M1, did not affect these functions. Arginine or leucine substitution of Asp59, located right at the bend of M1 seen in the crystal structure of the thapsigargin-bound form, caused a 10-fold increase of the rate of Ca2+ dissociation toward the cytoplasmic side. Mutation of Leu60 to alanine or proline and of Val62 to alanine also enhanced Ca2+ dissociation, whereas an 11-fold reduction of the rate of Ca2+ dissociation was observed upon alanine substitution of Leu65, thus providing evidence for a relation of the middle part of M1 to a gating mechanism controlling the dissociation of occluded Ca2+ from its membranous binding sites. Moreover, phosphoenzyme processing was affected by some of the latter mutations, in particular leucine substitution of Asp59, and alanine substitution of Leu65 accelerated the transition to ADP-insensitive phosphoenzyme and blocked its dephosphorylation, thus demonstrating that this part of M1, besides being important in Ca2+ interaction, furthermore, is a critical element in the long range signaling between the transmembrane domain and the cytoplasmic catalytic site. PMID- 14754884 TI - Role of the NH2 terminus in the assembly and trafficking of the intermediate conductance Ca2+-activated K+ channel hIK1. AB - The role of the NH(2)-terminal leucine zipper and dileucine motifs of hIK1 in the assembly, trafficking, and function of the channel was investigated using cell surface immunoprecipitation, co-immunoprecipitation (Co-IP), immunoblot, and whole-cell patch clamp techniques. Mutation of the NH(2)-terminal leucine zipper at amino acid positions 18 and 25 (L18A/L25A) resulted in a complete loss of steady-state protein expression, cell surface expression, and whole-cell current density. Inhibition of proteasomal degradation with lactacystin restored L18A/L25A protein expression, although this channel was not expressed at the cell surface as assessed by cell surface immunoprecipitation and whole-cell patch clamp. In contrast, inhibitors of lysosomal degradation (leupeptin/pepstatin) and endocytosis (chloroquine) had little effect on L18A/L25A protein expression or localization. Further studies confirmed the rapid degradation of this channel, having a time constant of 19.0 +/- 1.3 min compared with 3.2 +/- 0.8 h for wild type hIK1. Co-expression studies demonstrated that the L18A/L25A channel associates with wild type channel, thereby attenuating its expression at the cell surface. Co-IP studies confirmed this association. However, L18A/L25A channels failed to form homotetrameric channels, as assessed by Co-IP, suggesting the NH(2) terminus plays a role in tetrameric channel assembly. As with the leucine zipper, mutation of the dileucine motif to alanines, L18A/L19A, resulted in a near complete loss in steady-state protein expression with the protein being similarly targeted to the proteasome for degradation. In contrast to our results on the leucine zipper, however, both chloroquine and growing the cells at the permissive temperature of 27 degrees C restored expression of L18A/L19A at the cell surface, suggesting that the defect in the channel trafficking is the result of a subtle folding error. In conclusion, we demonstrate that the NH(2) terminus of hIK1 contains overlapping leucine zipper and dileucine motifs essential for channel assembly and trafficking to the plasma membrane. PMID- 14754885 TI - The N-terminal metal-binding site 2 of the Wilson's Disease Protein plays a key role in the transfer of copper from Atox1. AB - The Wilson's disease protein (WNDP) is a copper-transporting ATPase regulating distribution of copper in the liver. Mutations in WNDP lead to a severe metabolic disorder, Wilson's disease. The function of WNDP depends on Atox1, a cytosolic metallochaperone that delivers copper to WNDP. We demonstrate that the metal binding site 2 (MBS2) in the N-terminal domain of WNDP (N-WNDP) plays an important role in this process. The transfer of one copper from Atox1 to N-WNDP results in selective protection of the metal-coordinating cysteines in MBS2 against labeling with a cysteine-directed probe. Such selectivity is not observed when free copper is added to N-WNDP. Similarly, site-directed mutagenesis of MBS2 eliminates stimulation of the catalytic activity of WNDP by the copper-Atox1 complex but not by free copper. The Atox1 preference toward MBS2 is likely due to specific protein-protein interactions and is not due to unique surface exposure of the metal-coordinating residues or higher copper binding affinity of MBS2 compared with other sites. Competition experiments using a copper chelator revealed that MBS2 retained copper much better than Atox1, and this may facilitate the metal transfer process. X-ray absorption spectroscopy of the isolated recombinant MBS2 demonstrated that this sub-domain coordinates copper with a linear biscysteinate geometry, very similar to that of Atox1. Therefore, non-coordinating residues in the vicinity of the metal-binding sites are responsible for the difference in the copper binding properties of MBS2 and Atox1. The intramolecular changes that accompany transfer of a single copper to N WNDP are discussed. PMID- 14754886 TI - The Gtx homeodomain transcription factor exerts neuroprotection using its homeodomain. AB - Certain cases of familial Alzheimer's disease are caused by mutants of amyloid beta precursor protein (AbetaPP), including V642I-AbetaPP, K595N/M596L-AbetaPP (NL-AbetaPP), A617G-AbetaPP, and L648P-AbetaPP. By using an unbiased functional screening with transfection and expression of a human brain cDNA library, we searched for genes that protect neuronal cells from toxicity by V642I-AbetaPP. One protective clone was identical to the human GTX, a neuronal homeobox gene. Human Gtx (hGtx) inhibited caspase inhibitor-sensitive neuronal cell death not only by V642I-AbetaPP but also by L648P-, NL-, A617G-AbetaPP, apolipoprotein E4, and Abeta. The region of hGtx responsible for this rescue function was specified to be its homeodomain (Lys148-His207). The rescue function was shared by DLX4, a distal-less family gene with a homeodomain only 38.3% homologous to that of hGtx, suggesting that this function would be generally shared by homeodomains. The neuroprotective function of hGtx was attributable to hGtx-stimulated production and secretion of insulin-like growth factor-I. This study provides molecular clues to understand how neuronal cells developmentally regulate themselves against cell death as well as to develop reagents effective in curative therapeutics of Alzheimer's disease. PMID- 14754887 TI - Distinct natures of beryllium fluoride-bound, aluminum fluoride-bound, and magnesium fluoride-bound stable analogues of an ADP-insensitive phosphoenzyme intermediate of sarcoplasmic reticulum Ca2+-ATPase: changes in catalytic and transport sites during phosphoenzyme hydrolysis. AB - The structural natures of stable analogues for the ADP-insensitive phosphoenzyme (E2P) of Ca(2+)-ATPase formed in sarcoplasmic reticulum vesicles, i.e. the enzymes with bound beryllium fluoride (BeF.E2), bound aluminum fluoride (AlF.E2), and bound magnesium fluoride (MgF.E2), were explored and compared with those of actual E2P formed from P(i) without Ca(2+). Changes in trinitrophenyl-AMP fluorescence revealed that the catalytic site is strongly hydrophobic in BeF.E2 as in E2P but hydrophilic in MgF.E2 and AlF.E2; yet, the three cytoplasmic domains are compactly organized in these states. Thapsigargin, which was shown in the crystal structure to fix the transmembrane helices and, thus, the postulated Ca(2+) release pathway to lumen in a closed state, largely reduced the tryptophan fluorescence in BeF.E2 as in E2P, but only very slightly (hence, the release pathway is likely closed without thapsigargin) in MgF.E2 and AlF.E2 as in dephosphorylated enzyme. Consistently, the completely suppressed Ca(2+)-ATPase activity in BeF-treated vesicles was rapidly restored in the presence of ionophore A23187 but not in its absence by incubation with Ca(2+) (over several millimolar concentrations) at pH 6, and, therefore, lumenal Ca(2+) is accessible to reactivate the enzyme. In contrast, no or only very slow restoration was observed with vesicles treated with MgF and AlF even with A23187. BeF.E2 thus has the features very similar to those characteristic of the E2P ground state, although AlF.E2 and MgF.E2 most likely mimic the transition or product state for the E2P hydrolysis, during which the hydrophobic nature around the phosphorylation site is lost and the Ca(2+) release pathway is closed. The change in hydrophobic nature is probably associated with the change in phosphate geometry from the covalently bound tetrahedral ground state (BeF(3)(-)) to trigonal bipyramidal transition state (AlF(3) or AlF(4)(-)) and further to tetrahedral product state (MgF(4)(2-)), and such change likely rearranges transmembrane helices to prevent access and leakage of lumenal Ca(2+). PMID- 14754888 TI - Effects of different experimental conditions on the PrPSc core generated by protease digestion: implications for strain typing and molecular classification of CJD. AB - The discovery of molecular subtypes of the pathological prion protein PrPSc has provided the basis for a novel classification of human transmissible spongiform encephalopathies (TSEs) and a potentially powerful method for strain typing. However, there is still a significant disparity regarding the understanding and nomenclature of PrPSc types. In addition, it is still unknown whether a specific PrPSc type is associated with each TSE phenotypic variant. In sporadic Creutzfeldt-Jakob disease (sCJD), five disease phenotypes are known, but only two major types of PrPSc, types 1 and 2, have been consistently reproduced. We further analyzed PrPSc properties in sCJD and variant CJD using a high resolution gel electrophoresis system and varying experimental conditions. We found that pH varies among CJD brain homogenates in standard buffers, thereby influencing the characteristics of protease-treated PrPSc. We also show that PrPSc type 1 and type 2 are heterogeneous species which can be further distinguished into five molecular subtypes that fit the current histopathological classification of sCJD variants. Our results shed light on previous disparities in PrPSc typing, provide a refined classification of human PrPSc types, and support the notion that the pathological TSE phenotype is related to PrPSc structure. PMID- 14754889 TI - Squalestatin cures prion-infected neurons and protects against prion neurotoxicity. AB - A key feature of prion diseases is the conversion of the normal, cellular prion protein (PrP(C)) into beta-sheet-rich disease-related isoforms (PrP(Sc)), the deposition of which is thought to lead to neurodegeneration. In the present study, the squalene synthase inhibitor squalestatin reduced the cholesterol content of cells and prevented the accumulation of PrP(Sc) in three prion infected cell lines (ScN2a, SMB, and ScGT1 cells). ScN2a cells treated with squalestatin were also protected against microglia-mediated killing. Treatment of neurons with squalestatin resulted in a redistribution of PrP(C) away from Triton X-100 insoluble lipid rafts. These effects of squalestatin were dose-dependent, were evident at nanomolar concentrations, and were partially reversed by cholesterol. In addition, uninfected neurons treated with squalestatin became resistant to the otherwise toxic effect of PrP peptides, a synthetic miniprion (sPrP106) or partially purified prion preparations. The protective effect of squalestatin, which was reversed by the addition of water-soluble cholesterol, correlated with a reduction in prostaglandin E(2) production that is associated with neuronal injury in prion disease. These studies indicate a pivotal role for cholesterol-sensitive processes in controlling PrP(Sc) formation, and in the activation of signaling pathways associated with PrP-induced neuronal death. PMID- 14754890 TI - Identification of the N-terminal peptide binding site of glucose-regulated protein 94. AB - Because the stress protein GRP94 can augment presentation of peptides to T cells, it is important to define how it, as well as all other HSP90 family members, binds peptides. Having previously shown that the N-terminal half of GRP94 can account for the peptide binding activity of the full-length protein, we now locate this binding site by testing predictions of a molecular docking model. The best predicted site was on the opposite face of the beta sheet from the pan-HSP90 radicicol-binding pocket, in close proximity to a deep hydrophobic pocket. The peptide and radicicol-binding sites are distinct, as shown by the ability of a radicicol-refractive mutant to bind peptide. When the fluorophore acrylodan is attached to Cys117 within the hydrophobic pocket, its fluorescence is reduced upon peptide binding, consistent with proximity of the two ligands. Substitution of His125, which contacts the bound peptide, compromises peptide-binding activity. We conclude that peptide binds to the concave face of the beta sheet of the N-terminal domain, where binding is regulated during the action cycle of the chaperone. PMID- 14754891 TI - "Network leaning" as a mechanism of insurmountable antagonism of the angiotensin II type 1 receptor by non-peptide antagonists. AB - A mechanistic understanding of the insurmountable antagonism of the angiotensin II type 1 (AT(1)) receptor could be fundamental in the quest for discovery and improvement of drugs. Candesartan and EXP3174 are competitive, reversible insurmountable antagonists of the AT(1) receptor. They contain di-acidic substitutions, whereas the surmountable antagonist, losartan, contains only one acidic group. We tested the hypothesis that these two classes of ligands interact with the AT(1) receptor through similar but not identical bonds and that the differences in the acid-base group contacts are critical for insurmountable antagonism. By pharmacological characterization of site-directed AT(1) receptor mutants expressed in COS1 cells we show that specific interactions with Gln(257) in transmembrane 6 distinguishes insurmountable antagonists and that abolishing these interactions transforms insurmountable to surmountable antagonism. In the Q257A mutant, the dissociation rate of [(3)H]candesartan is 2.8-fold more than the rate observed with wild type, and the association rate was reduced 4-fold lower than the wild type. The pattern of antagonism of angiotensin II concentration-response in the Q257A mutant pretreated with EXP3174 and candesartan is surmountable. We propose that leaning ability of insurmountable antagonists on Gln(257) in the wild-type receptor is the basis of an antagonist mediated conformational transition, which is responsible for both slow dissociation and inhibition of maximal IP response. PMID- 14754892 TI - Regulation of cyclin D1/Cdk4 complexes by calcium/calmodulin-dependent protein kinase I. AB - The selective inhibitor of the multifunctional calcium/calmodulin-dependent kinases (CaMK), KN-93, arrests a variety of cell types in G(1). However, the biochemical nature of this G(1) arrest point and the physiological target of KN 93 in G(1) remain controversial. Here we show that in WI-38 human diploid fibroblasts KN-93 reversibly arrested cells in late G(1) prior to detectable cyclin-dependent kinase 4 (cdk4) activation. At the KN-93 arrest point, we found that cyclin D1/cdk4 complexes had assembled with p21/p27, accumulated in the nucleus, and become phosphorylated on Thr-172, yet were relatively inactive. Additional examination of cdk4 complexes by gel filtration analysis demonstrated that, in late G(1), cyclin D1-containing complexes migrated toward lower molecular weight (M(r)) fractions and this altered migration was accompanied by the appearance of two peaks of cdk4 activity, at 150-200 and 70 kDa, respectively. KN-93 prevented both the activation of cdk4, and this shift in cyclin D1 migration and overexpression of cyclin D1/cdk4 overcame the KN-93 arrest. To determine which multifunctional CaMK acts in G(1), we expressed kinase deficient forms of CaMKI and CaMKII. Overexpression of kinase-deficient CaMKI, but not CaMKII, prevented cdk4 activation, mimicking the KN-93 arrest point. Therefore, we hypothesize that KN-93 prevents a very late, uncharacterized step in cyclin D/cdk4 activation that involves CaMKI and follows complex assembly, nuclear entry, and phosphorylation. PMID- 14754893 TI - PE-1/METS, an antiproliferative Ets repressor factor, is induced by CREB-1/CREM-1 during macrophage differentiation. AB - The molecular mechanisms involved in regulating the balance between cellular proliferation and differentiation remain poorly understood. Members of the Ets domain family of transcription factors are candidates for proteins that might differentially regulate cell cycle control and cell type-specific genes during the differentiation of myeloid progenitor cells. The Ets repressor PE-1/METS has been suggested to contribute to growth arrest during terminal macrophage differentiation by repressing Ets target genes involved in Ras-dependent proliferation. An important feature of this regulatory model is that PE-1/METS is itself induced by the program of macrophage differentiation elicited by M-CSF. Here, we present evidence that the PE-1/METS gene is a transcriptional target of the cyclic AMP response element-binding protein-1 (CREB-1). CREB-1 expression is dramatically up-regulated during macrophage differentiation and phosphorylation of CREB-1 and the related factor CREM-1 are stimulated by M-CSF in a SAPK2/p38 dependent manner. Chromatin immunoprecipitation experiments demonstrate that CREB 1/CREM-1 are recruited to the PE-1/METS promoter as well as to the promoters of other genes that are up-regulated during terminal macrophage differentiation. Overexpression of CREB-1 stimulates the activities of the PE-1/METS, and macrosialin promoters, while expression of a dominant negative form of CREB-1 during macrophage differentiation inhibits expression of the PE-1/METS and macrosialin genes. Inhibition of CREB function also results in reduced expression of CD54 and impaired cell adhesion. Taken together, these findings reveal new roles of CREB-1/CREM-1 as regulators of macrophage differentiation. PMID- 14754894 TI - cAMP-induced astrocytic differentiation of C6 glioma cells is mediated by autocrine interleukin-6. AB - Elevation in the level of intracellular cAMP is known to induce the astrocytic differentiation of C6 glioma cells by unknown mechanisms. In this report, we show that cAMP-induced autocrine interleukin 6 (IL-6) promoted astrocytic differentiation of C6 cells. Treatment of cells with N(6),2'-O-dibutyryl cAMP (Bt(2)AMP) and theophylline caused the delayed phosphorylation of signal transducer and activator of transcription 3 (STAT3), as well as the expression of an astrocyte marker, glial fibrillary acidic protein (GFAP). Overexpression of the dominant-negative form of STAT3 leads to the suppression of GFAP promoter activity, suggesting that STAT3 activity was essential for cAMP-induced GFAP promoter activation. On the other hand, the IL-6 gene was quickly induced by Bt(2)AMP/theophylline, and subsequent IL-6 protein secretion was stimulated. In addition, recombinant IL-6 induced GFAP expression and STAT3 phosphorylation. Most importantly, treatment with IL-6-neutralizing antibody dramatically reduced the cAMP-induced GFAP expression and STAT3 phosphorylation and reversed the cellular morphological changes that had been caused by Bt(2)AMP/theophylline. Taken together, these results indicated that Bt(2)AMP/theophylline lead to delayed STAT3 activation via autocrine IL-6. These processes subsequently led to the induction of GFAP. IL-6 secretion is thus thought to be a key event in controlling the astrocytic differentiation of C6 cells. PMID- 14754895 TI - ACE2 X-ray structures reveal a large hinge-bending motion important for inhibitor binding and catalysis. AB - The angiotensin-converting enzyme (ACE)-related carboxypeptidase, ACE2, is a type I integral membrane protein of 805 amino acids that contains one HEXXH + E zinc binding consensus sequence. ACE2 has been implicated in the regulation of heart function and also as a functional receptor for the coronavirus that causes the severe acute respiratory syndrome (SARS). To gain further insights into this enzyme, the first crystal structures of the native and inhibitor-bound forms of the ACE2 extracellular domains were solved to 2.2- and 3.0-A resolution, respectively. Comparison of these structures revealed a large inhibitor-dependent hinge-bending movement of one catalytic subdomain relative to the other ( approximately 16 degrees ) that brings important residues into position for catalysis. The potent inhibitor MLN-4760 ((S,S)-2-[1-carboxy-2-[3-(3,5 dichlorobenzyl)-3H-imidazol4-yl]-ethylamino]-4-methylpentanoic acid) makes key binding interactions within the active site and offers insights regarding the action of residues involved in catalysis and substrate specificity. A few active site residue substitutions in ACE2 relative to ACE appear to eliminate the S(2)' substrate-binding subsite and account for the observed reactivity change from the peptidyl dipeptidase activity of ACE to the carboxypeptidase activity of ACE2. PMID- 14754896 TI - MALT1/paracaspase is a signaling component downstream of CARMA1 and mediates T cell receptor-induced NF-kappaB activation. AB - T cell receptor (TCR) induces a series of signaling cascades and leads to activation of multiple transcription factors, including NF-kappaB. Although the mechanism of TCR-induced NF-kappaB activation is not fully understood, recent studies indicate that Bcl10 and CARMA1, two adaptor/scaffold proteins, play essential roles in mediating TCR-induced NF-kappaB activation. MALT1/paracaspase is a caspase-like protein that contains an N-terminal death domain, two Ig-like domains, and a C-terminal caspase-like domain. It binds to Bcl10 through its Ig like domains and cooperates with Bcl10 to activate NF-kappaB. Recently, it has been shown that MALT1 is involved in mediating TCR signal transduction, leading to activation of NF-kappaB. In this study, we show that MALT1 is recruited into the lipid rafts of the immunological synapse following activation of the TCR and the CD28 coreceptor (CD3/CD28 costimulation). This recruitment of MALT1 is dependent on CARMA1 because CD3/CD28 costimulation failed to recruit MALT1 into lipid rafts in CARMA1-deficient T cells. In addition, we also found that MALT1 not only binds to Bcl10 directly, but also associates with CARMA1 in a Bcl10 independent manner. Therefore, MALT1, Bcl10, and CARMA1 form a trimolecular complex. Expression of a MALT1 deletion mutant containing only the N-terminal death domain and the two Ig-like domains completely blocked CD3/CD28 costimulation-induced, but not tumor necrosis factor-alpha-induced, NF-kappaB activation. Together, these results indicate that MALT1 is a crucial signaling component in the TCR signaling pathway. PMID- 14754897 TI - ZNF216 Is an A20-like and IkappaB kinase gamma-interacting inhibitor of NFkappaB activation. AB - The transcription factor NFkappaB plays important roles in immune regulation, inflammatory responses, and anti-apoptosis. Activation of NFkappaB requires the activity of IkappaB kinase, a kinase complex that contains two catalytic subunits, IKKalpha and IKKbeta, and a non-enzymatic regulatory subunit, IKKgamma. To understand how NFkappaB activation is regulated at the IKKgamma level, we searched for IKKgamma-interacting proteins by the yeast two-hybrid system. This search identified ZNF216, a zinc finger protein with unknown biological functions. ZNF216 contains an A20-like zinc finger domain (ZnF-A20) at its N terminus and an AN1-like domain (ZnF-AN1) at its C terminus. Similar to A20, ZNF216 interacted with IKKgamma, RIP, and TRAF6 in co-immunoprecipitation experiments. Domain mapping experiments indicated that the ZnF-A20 domain was responsible for interacting with IKKgamma and RIP, whereas the ZnF-AN1 domain interacted with TRAF6. ZNF216 inhibited NFkappaB activation triggered by overexpression of RIP and TRAF6 but not of p65. ZNF216 also inhibited tumor necrosis factor (TNF)-, interleukin-1-, and Toll-like receptor 4-induced NFkappaB activation in a dose-dependent manner. The ZnF-A20 domain was essential for ZNF216-mediated inhibition of NFkappaB activation. The ZnF-A20 and ZnF-AN1 domains of ZNF216 could interact with each other, whereas ZNF216 could form homo oligomers or hetero-oligomers with A20. Unlike A20, which inhibits TNF-induced apoptosis, overexpression of ZNF216 sensitized cells to TNF-induced apoptosis. Our findings suggest that ZNF216 and A20 have redundant and distinct roles in regulating NFkappaB activation and apoptosis. PMID- 14754898 TI - Directed in vitro evolution and crystallographic analysis of a peptide-binding single chain antibody fragment (scFv) with low picomolar affinity. AB - We generated a single chain Fv fragment of an antibody (scFv) with a binding affinity of about 5 pm to a short peptide by applying rigorous directed evolution. Starting from a high affinity peptide binder, originally obtained by ribosome display from a murine library, we generated libraries of mutants with error-prone PCR and DNA shuffling and applied off-rate selection by using ribosome display. Crystallographic analysis of the scFv in its antigen-bound and free state showed that only few mutations, which do not make direct contact to the antigen, lead to a 500-fold affinity improvement over its potential germ line precursor. These results suggest that the affinity optimization of very high affinity binders is achieved by modulating existing interactions via subtle changes in the framework rather than by introducing new contacts. Off-rate selection in combination with ribosome display can evolve binders to the low picomolar affinity range even for peptide targets. PMID- 14754899 TI - Cathepsin L1, the major protease involved in liver fluke (Fasciola hepatica) virulence: propetide cleavage sites and autoactivation of the zymogen secreted from gastrodermal cells. AB - The secretion and activation of the major cathepsin L1 cysteine protease involved in the virulence of the helminth pathogen Fasciola hepatica was investigated. Only the fully processed and active mature enzyme can be detected in medium in which adult F. hepatica are cultured. However, immunocytochemical studies revealed that the inactive procathepsin L1 is packaged in secretory vesicles of epithelial cells that line the parasite gut. These observations suggest that processing and activation of procathepsin L1 occurs following secretion from these cells into the acidic gut lumen. Expression of the 37-kDa procathepsin L1 in Pichia pastoris showed that an intermolecular processing event within a conserved GXNXFXD motif in the propeptide generates an active 30-kDa intermediate form. Further activation of the enzyme was initiated by decreasing the pH to 5.0 and involved the progressive processing of the 37 and 30-kDa forms to other intermediates and finally to a fully mature 24.5 kDa cathepsin L with an additional 1 or 2 amino acids. An active site mutant procathepsin L, constructed by replacing the Cys(26) with Gly(26), failed to autoprocess. However, [Gly(26)]procathepsin L was processed by exogenous wild-type cathepsin L to a mature enzyme plus 10 amino acids attached to the N terminus. This exogenous processing occurred without the formation of a 30-kDa intermediate form. The results indicate that activation of procathepsin L1 by removal of the propeptide can occur by different pathways, and that this takes place within the parasite gut where the protease functions in food digestion and from where it is liberated as an active enzyme for additional extracorporeal roles. PMID- 14754902 TI - Integrin activation. AB - The ability of cells to regulate dynamically their adhesion to one another and to the extracellular matrix (ECM) that surrounds them is essential in multicellular organisms. The integrin family of transmembrane adhesion receptors mediates both cell-cell and cell-ECM adhesion. One important, rapid and reversible mechanism for regulating adhesion is by increasing the affinity of integrin receptors for their extracellular ligands (integrin activation). This is controlled by intracellular signals that, through their action on integrin cytoplasmic domains, induce conformational changes in integrin extracellular domains that result in increased affinity for ligand. Recent studies have shed light on the final intracellular steps in this process and have revealed a vital role for the cytoskeletal protein talin. PMID- 14754903 TI - Fibroblast heterogeneity: more than skin deep. AB - Dermal fibroblasts are a dynamic and diverse population of cells whose functions in skin in many respects remain unknown. Normal adult human skin contains at least three distinct subpopulations of fibroblasts, which occupy unique niches in the dermis. Fibroblasts from each of these niches exhibit distinctive differences when cultured separately. Specific differences in fibroblast physiology are evident in papillary dermal fibroblasts, which reside in the superficial dermis, and reticular fibroblasts, which reside in the deep dermis. Both of these subpopulations of fibroblasts differ from the fibroblasts that are associated with hair follicles. Fibroblasts engage in fibroblast-epidermal interactions during hair development and in interfollicular regions of skin. They also play an important role in cutaneous wound repair and an ever-increasing role in bioengineering of skin. Bioengineered skin currently performs important roles in providing (1) a basic understanding of skin biology, (2) a vehicle for testing topically applied products and (3) a resource for skin replacement. PMID- 14754904 TI - Nuclear translocation of the Hsp70/Hsp90 organizing protein mSTI1 is regulated by cell cycle kinases. AB - The co-chaperone murine stress-inducible protein 1 (mSTI1), an Hsp70/Hsp90 organizing protein (Hop) homologue, mediates the assembly of the Hsp70/Hsp90 chaperone heterocomplex. The mSTI1 protein can be phosphorylated in vitro by cell cycle kinases proximal to a putative nuclear localization signal (NLS), which substantiated a predicted casein kinase II (CKII)-cdc2 kinase-NLS (CcN) motif at position 180-239 and suggested that mSTI1 might move between the cytoplasm and the nucleus under certain cell cycle conditions. The mechanism responsible for the cellular localization of mSTI1 was probed using NIH3T3 fibroblasts to investigate the localization of endogenous mSTI1 and enhanced green fluorescent protein (EGFP)-tagged mSTI1 mutants. Localization studies on cell lines stably expressing NLS(mSTI1)-EGFP and EGFP demonstrated that the NLS(mSTI1) was able to promote a nuclear localization of EGFP. The mSTI1 protein was exclusively cytoplasmic in most cells under normal conditions but was present in the nucleus of a subpopulation of cells and accumulated in the nucleus following inhibition of nuclear export (leptomycin B treatment). G1/S-phase arrest (using hydroxyurea) and inhibition of cdc2 kinase (using olomoucine) but not inhibition of casein kinase II (using 5,6-dichlorobenzimidazole riboside), increased the proportion of cells with endogenous mSTI1 nuclear staining. mSTI1-EGFP behaved identically to endogenous mSTI1. The functional importance of key residues was tested using modified mSTI1-EGFP proteins. Inactivation and phosphorylation mimicking of potential phosphorylation sites in mSTI1 altered the nuclear translocation. Mimicking of phosphorylation at the mSTI1 CKII phosphorylation site (S189E) promoted nuclear localization of mSTI1-EGFP. Mimicking phosphorylation at the cdc2 kinase phosphorylation site (T198E) promoted cytoplasmic localization of mSTI1-EGFP at the G1/S-phase transition,whereas removal of this site (T198A) promoted the nuclear localization of mSTI1-EGFP under the same conditions. These data provide the first evidence of nuclear import and export of a major Hsp70/Hsp90 co-chaperone and the regulation of this nuclear-cytoplasmic shuttling by cell cycle status and cell cycle kinases. PMID- 14754905 TI - A simple, rapid, and sensitive fluorescence assay for microsomal triglyceride transfer protein. AB - Microsomal triglyceride transfer protein (MTP) is critical for the assembly and secretion of apolipoprotein B (apoB) lipoproteins. Its activity is classically measured by incubating purified MTP or cellular homogenates with donor vesicles containing radiolabeled lipids, precipitating the donor vesicles, and measuring the radioactivity transferred to acceptor vesicles. Here, we describe a simple, rapid, and sensitive fluorescence assay for MTP. In this assay, purified MTP or cellular homogenates are incubated with small unilamellar donor vesicles containing quenched fluorescent lipids (triacylglycerols, cholesteryl esters, and phospholipids) and different types of acceptor vesicles made up of phosphatidylcholine or phosphatidylcholine and triacylglycerols. Increases in fluorescence attributable to MTP-mediated lipid transfer are measured after 30 min. MTP's lipid transfer activity could be assayed using apoB lipoproteins but not with high density lipoproteins as acceptors. The assay was used to measure MTP activity in cell and tissue homogenates. Furthermore, the assay was useful in studying the inhibition of the cellular as well as purified MTP by its antagonists. This new method is amenable to automation and can be easily adopted for large-scale, high-throughput screening. PMID- 14754906 TI - Modification of LDL with human secretory phospholipase A(2) or sphingomyelinase promotes its arachidonic acid-releasing propensity. AB - Oxidation and lipolytic remodeling of LDL are believed to stimulate LDL entrapment in the arterial wall, expanding the inflammatory response and promoting atherosclerosis. However, the cellular responses and molecular mechanisms underlying the atherogenic effects of lipolytically modified LDL are incompletely understood. Human THP-1 monocytes were prelabeled with [(3)H]arachidonic acid (AA) before incubation with LDL or LDL lipolytically modified by secretory PLA(2) (sPLA(2)) or bacterial sphingomyelinase (SMase). LDL elicited rapid and dose-dependent extracellular release of AA in monocytes. Interestingly, LDL modified by sPLA(2) or SMase displayed a marked increase in AA mobilization relative to native LDL, and this increase correlated with enhanced activity of cytosolic PLA(2) (cPLA(2)) assayed in vitro as well as increased monocyte tumor necrosis factor-alpha secretion. The AA liberation was attenuated by inhibitors toward cPLA(2) and sPLA(2), indicating that both PLA(2) enzymes participate in LDL-induced AA release. In conclusion, these results demonstrate that LDL lipolytically modified by sPLA(2) or SMase potentiates cellular AA release and cPLA(2) activation in human monocytes. From our results, we suggest novel atherogenic properties for LDL modified by sPLA(2) and SMase in AA release and signaling, which could contribute to the inflammatory gene expression observed in atherosclerosis. PMID- 14754907 TI - Induction of lysosomal phospholipase A2 through the retinoid X receptor in THP-1 cells. AB - An acidic phospholipase A(2) (LPLA(2)) was recently purified and cloned. THP-1 cells were used to characterize the gene induction of LPLA(2). THP-1 cells were stimulated with several differentiation agents. The LPLA(2) mRNA and activity increased in cells treated with phorbol ester but not with vitamin D3, interferon gamma, or granulocyte macrophage colony-stimulating factor. All-trans-retinoic acid enhanced mRNA expression and enzyme activity in a dose- and time-dependent manner. The natural 9-cis and 13-cis isomers of retinoic acid enhanced transcription and activity. Two classes of nuclear receptors, the retinoic acid receptor (RAR) and the retinoid X receptor (RXR), mediate retinoic acid signaling. Specific RAR and RXR agonists were used to identify the nuclear receptor responsible for LPLA(2) induction by retinoic acid. Treatment with the RAR agonist 4-[E-2-tetrahydro-5,5,8,8-tetra-methyl-2-naphthalenyl]1-propenyl benzoic acid (TTNPB) resulted in a small and statistically significant increase of the mRNA expression and activity of LPLA(2). The RXR agonist methoprene acid worked as well as all-trans-retinoic acid at increasing both mRNA and enzyme activity. The methoprene acid and TTNPB effects were not synergistic. The peroxisome proliferator-activated receptor gamma agonists 15-deoxy-Delta(12,14) prostaglandin J(2) and troglitazone failed to induce LPLA(2) activity and mRNA. Thus, an RXR-dependent pathway controls LPLA(2) gene activation by retinoic acid in THP-1 cells. PMID- 14754908 TI - Molecular interactions between apoE and ABCA1: impact on apoE lipidation. AB - Apolipoprotein E (apoE)/ABCA1 interactions were investigated in human intact fibroblasts induced with 22(R)-hydroxycholesterol and 9-cis-retinoic acid (stimulated cells). Here, we show that purified human plasma apoE3 forms a complex with ABCA1 in normal fibroblasts. Lipid-free apoE3 inhibited the binding of (125)I-apoA-I to ABCA1 more efficiently than reconstituted HDL particles (IC(50) = 2.5 +/- 0.4 microg/ml vs. 12.3 +/- 1.3 microg/ml). ApoE isoforms showed similar binding for ABCA1 and exhibited identical kinetics in their abilities to induce ABCA1-dependent cholesterol efflux. Mutation of ABCA1 associated with Tangier disease (C1477R) abolished both apoE3 binding and apoE3-mediated cholesterol efflux. Analysis of apoE3-containing particles generated during the incubation of lipid-free apoE3 with stimulated normal cells showed nascent apoE3/cholesterol/phospholipid complexes that exhibited prebeta-electrophoretic mobility with a particle size ranging from 9 to 15 nm, whereas lipid-free apoE3 incubated with ABCA1 mutant (C1477R) cells was unable to form such particles. These results demonstrate that 1). apoE association with lipids reduced its ability to interact with ABCA1; 2). apoE isoforms did not affect apoE binding to ABCA1; 3). apoE-mediated ABCA1-dependent cholesterol efflux was not affected by apoE isoforms in fibroblasts; and 4). the lipid translocase activity of ABCA1 generates apoE-containing high density-sized lipoprotein particles. Thus, ABCA1 is essential for the biogenesis of high density-sized lipoprotein containing only apoE particles in vivo. PMID- 14754909 TI - Incorporation of cis-9,trans-11 or trans-10,cis-12 conjugated linoleic acid into plasma and cellular lipids in healthy men. AB - This study investigated the incorporation of cis-9,trans-11 conjugated linoleic acid (c9,t11 CLA) and trans-10,cis-12-CLA (t10,c12 CLA) into plasma and peripheral blood mononuclear cell (PBMC) lipids when consumed as supplements highly enriched in these isomers. Healthy men (n = 49, age 31 +/- 8 years) consumed one, two, and four capsules containing approximately 600 mg of either c9,t11 CLA or t10,c12 CLA per capsule for sequential 8 week periods followed by a 6 week washout before consuming the alternative isomer. Both isomers were incorporated in a dose-dependent manner into plasma phosphatidylcholine (PC) (c9,t11 CLA r = 0.779, t10,c12 CLA r = 0.738; P < 0.0001) and cholesteryl ester (CE) (c9,t11 CLA r = 0.706, t10,c12 CLA r = 0.788; P < 0.0001). Only t10,c12 CLA was enriched in plasma nonesterified fatty acids. Both c9,t11 CLA and t10,c12 CLA were incorporated linearly into PBMC total lipids (r = 0.285 and r = 0.273, respectively; P < 0.0005). The highest concentrations of c9,t11 CLA and t10,c12 CLA in PBMC lipids were 3- to 4-fold lower than those in plasma PC and CE. These data suggest that the level of intake is a major determinant of plasma and PBMC CLA content, although PBMCs appear to incorporate both CLA isomers less readily. PMID- 14754910 TI - Human lipoproteins have divergent neutralizing effects on E. coli LPS, N. meningitidis LPS, and complete Gram-negative bacteria. AB - The use of lipoproteins has been suggested as a treatment for Gram-negative sepsis because they inhibit lipopolysaccharide (LPS)-mediated cytokine production. However, little is known about the neutralizing effects of lipoproteins on cytokine production by meningococcal LPS or whole Gram-negative bacteria. We assessed the neutralizing effect of LDLs, HDLs, and VLDLs on LPS- or whole bacteria-induced cytokines in human mononuclear cells. A strong inhibition of Escherichia coli LPS-induced interleukin-1beta (IL-1beta), tumor necrosis factor-alpha, and IL-10 by LDL and HDL was seen, whereas VLDL had a less pronounced effect. In contrast, Neisseria meningitidis LPS, in similar concentrations, was neutralized much less effectively than E. coli LPS. Effective neutralization of meningococcal LPS required a longer interaction time, a lower concentration of LPS, or higher concentrations of lipoproteins. The difference in neutralization was independent of the saccharide tail, suggesting that the lipid A moiety accounted for the difference. Minimal neutralizing effects of the lipoproteins were observed on whole E. coli or N. meningitidis bacteria under all conditions tested. These results indicate that efficient neutralization of LPS depends on the type of LPS, but a sufficiently long interaction time, a low LPS concentration, or high lipoprotein concentration also inhibited cytokines by the less efficiently neutralized N. meningitidis LPS. Irrespective of these differences, whole bacteria showed no neutralization by lipoproteins. PMID- 14754911 TI - Synthesis and metabolism of leukotrienes in gamma-glutamyl transpeptidase deficiency. AB - Leukotrienes (LTs) are active lipid mediators derived in the 5-lipoxygenase pathway. LTC(4), the primary cysteinyl LT, is cleaved by gamma-glutamyl transpeptidase (GGT), resulting in LTD(4). We studied the synthesis and metabolism of LTs in three patients with GGT deficiency. LTs were analyzed in urine, plasma, and monocytes after HPLC separation by enzyme immunoassays, radioactivity detection, and electrospray tandem mass spectrometry. Analysis of LTs in urine revealed increased concentrations of LTC(4) (12.8-17.9 nmol/mol creatinine; controls, <0.005 nmol/mol creatinine), whereas LTE(4) was below the detection limit (<0.005 nmol/mol creatinine; controls, 32.2 +/- 8.6 nmol/mol creatinine). In plasma of one patient, LTC(4) was found to be increased (17.3 ng/ml; controls, 9.6 +/- 0.4 ng/ml), whereas LTD(4) and LTE(4) were below the detection limit (<0.005 ng/ml). LTB(4) was found within normal ranges. In contrast to controls, the synthesis of LTD(4) and LTE(4) in stimulated monocytes was below the detection limit (<0.1 ng/10(6) cells; controls, 37.1 +/- 4.8 cells and 39.4 +/- 5.6 ng/10(6) cells, respectively). The formation of [(3)H]LTD(4) from [(3)H]LTC(4) in monocytes was completely deficient (<0.1%; controls, 85 +/- 7%). Our data demonstrate a complete deficiency of LTD(4) biosynthesis in patients with a genetic deficiency of GGT. GGT deficiency represents a new inborn error of cysteinyl LT synthesis and provides a unique model in which to study the pathobiological coherence of LT and glutathione metabolism. PMID- 14754912 TI - Genetic determinants of obesity-related lipid traits. AB - In our ongoing effort to identify genes influencing the biological pathways that underlie the metabolic disturbances associated with obesity, we performed genome wide scanning in 2,209 individuals distributed over 507 Caucasian families to localize quantitative trait loci (QTLs), which affect variation of plasma lipids. Pedigree-based analysis using a quantitative trait variance component linkage method that localized a QTL on chromosome 7q35-q36, which linked to variation in levels of plasma triglyceride [TG, logarithm of odds (LOD) score = 3.7] and was suggestive of linkage to LDL-cholesterol (LDL-C, LOD = 2.2). Covariates of the TG linkage included waist circumference, fasting insulin, and insulin:glucose, but not body mass index or hip circumference. Plasma HDL-cholesterol (HDL-C) levels were suggestively linked to a second QTL on chromosome 12p12.3 (LOD = 2.6). Five other QTLs with lower LOD scores were identified for plasma levels of LDL-C, HDL C, and total cholesterol. These newly identified loci likely harbor genetic elements that influence traits underlying lipid adversities associated with obesity. PMID- 14754913 TI - Plasma phosphatidylcholine hydroperoxide as a new marker of oxidative stress in alcoholic patients. AB - Quantitative analysis of plasma phosphatidylcholine hydroperoxide (PCOOH) is an important step in evaluating the biochemical processes leading to oxidative injury. However, secondary products of lipid peroxidation are now used as indices. One hundred nine alcoholic patients, aged 22-81 years (mean +/- SEM, 52.0 +/- 1.3 years), and 21 healthy volunteers, aged 41-79 years (51.2 +/- 2.2 years), participated in this study. Plasma PCOOH was measured by HPLC with chemiluminescence detection. Plasma PCOOH concentration was significantly higher in alcoholic patients (46.1 +/- 4.1 pmol/ml) than in controls (15.6 +/- 1.8 pmol/ml). It was significantly higher in patients with blood alcohol (88.0 +/- 10.5 pmol/ml) than in those without alcohol (32.6 +/- 3.1 pmol/ml). The patients with high levels of aspartate aminotransferase, alanine aminotransferase, gamma glutamyl transpeptidase (gamma-GTP), and triglyceride (TG) showed significantly higher PCOOH concentrations than did patients with normal levels. The PCOOH level was positively correlated with levels of gamma-GTP, HDL, blood alcohol concentration, and TG. Plasma PCOOH levels in 29 alcoholic patients after a 6 week abstinence were decreased significantly (22.8 +/- 11.1 pmol/ml), which was associated with improvement on liver function tests. This is the first measurement of plasma PCOOH in alcoholic patients. These results suggest the involvement of lipid peroxidation in alcohol-induced liver damage and confirm that the PCOOH plasma concentration is a new marker of alcohol consumption as well as oxidative stress in alcoholic patients. PMID- 14754914 TI - The unusual vascular structure of the corm of Eriophorum vaginatum: implications for efficient retranslocation of nutrients. AB - Eriophorum spp. are abundant perennial graminoids in the Arctic tundra and boreal peatlands. Because ecological studies indicated that some plants are unusually productive on infertile and cold sites, the anatomy of the overwintering corms of Eriophorum vaginatum (L.) and Eriophorum scheuchzeri (Hoppe) were examined to determine their involvement in nutrient uptake and storage. Components of the long-distance transport pathways were identified within the plants by using histochemical techniques and transport of apoplastic and symplastic dyes. E. scheuchzeri produced a rhizome that consisted mainly of storage parenchyma cells within which collateral vascular bundles were centrally located and arranged in a circle. By contrast, E. vaginatum developed a ring of horizontally arranged xylem and phloem, in addition to axial amphivasal vascular bundles leading to the leaves, all of which were bordered by transfer cells. As shown by the transport of fluorescein in the phloem and Safranine O in the xylem, each axial bundle and adventitious root contacted the horizontal ring of vascular tissues so that solutes from one vascular bundle were translocated into the vascular ring and circulated to another vascular bundle and/or to the roots. In addition, special groups of sclereids that functioned in both phloem and xylem transport were found at the base of the leaf traces and within junctions of senescing roots. These sclereids were named 'vascular sclerenchyma' and it was hypothesized that they provide a moving end for the vascular system because the corm dies progressively from the distal end as it grows upward from the apical meristem. It was concluded that this unusual vascular system of E. vaginatum is efficient in recycling nutrients internally, which may account for its competitive advantage in infertile and cold sites. PMID- 14754915 TI - Differential expression of three genes encoding an ethylene receptor in rice during development, and in response to indole-3-acetic acid and silver ions. AB - Five ethylene receptor genes, OS-ERS1, OS-ERS2, OS-ETR2, OS-ETR3, and OS-ETR4 were isolated and characterized from rice. The genomic structure of OS-ERS1 and OS-ERS2 revealed that the introns within the coding sequences occurred in conserved positions to those of At-ETR1 and At-ERS1, whereas each of the OS-ETR2, OS-ETR3, and OS-ETR4 genes contained 1 intron within its coding region located at a position equivalent to those of At-ERS2, At-ETR2, and At-EIN4. Deduced amino acid sequences of OS-ERS1, OS-ERS2, OS-ETR2, OS-ETR3, and OS-ETR4 showed that they exhibited significant homology to the prokaryotic two-component signal transducer and a wide range of ethylene receptors in a variety of plant species. Northern analysis revealed that the level of OS-ETR2 mRNA was markedly elevated either by the exogenous application of IAA or by ethylene treatment in young etiolated rice seedlings, whereas the OS-ERS1 transcript level was only slightly induced under the same experimental conditions. Pretreatment with silver prevented IAA-induced and ethylene-induced accumulation of both mRNAs (OS-ERS1 and OS-ETR2). However, the abundance of OS-ERS2 mRNA was shown to be down regulated by both IAA and ethylene treatments, indicating that it was not positively regulated by ethylene. Analysis of the expression of the three ethylene receptor genes in different tissues of rice has unravelled their corresponding tissue-specificity in which OS-ERS1 was constitutively expressed in considerable amounts in all tissues studied, while OS-ERS2 and OS-ETR2 exhibited differential expression patterns in different tissues of rice. Moreover, higher levels of these three mRNAs were commonly observed in anthers when compared with their corresponding levels in other tissues, suggesting the important role played by ethylene involved in the regulation of pollen development in rice. Among the five ethylene receptor genes, the expression levels of both OS-ETR3 and OS-ETR4 were too low to be detected by the northern blot analysis. Results from RT-PCR illustrated that both mRNAs were present in young green rice seedlings and anthers. PMID- 14754916 TI - Interspecific control of non-symbiotic carbon partitioning in the rhizosphere of a grass-clover association: Bromus madritensis-Trifolium angustifolium. AB - Grass-legume interaction in the rhizosphere was investigated in a greenhouse experiment with two annual species, bromegrass Bromus madritensis (L.) and clover Trifolium angustifolium (L.) grown in mono and mixed cultures. Partitioning of below-ground carbon between roots, respiration, and soil was measured after separate 2 h-labelling of each species with 14CO2 followed by a 9 d chase period. At the time of labelling, clover nodules were not yet fixing N2. Bromegrass grew much faster than clover. Shoot biomass of bromegrass was greater in the presence of clover than in monoculture. By contrast, both shoot and root biomass of clover was less in the presence of bromegrass than in monoculture. Carbon assimilation during the period of labelling was proportional to shoot biomass and partitioning above and below-ground did not differ among treatments. Absolute amounts of labelled C allocated to rhizosphere respiration was more in bromegrass than in clover (respectively 1.38 mg C against 0.75 mg C in monoculture and 1.79 mg C and 0.63 mg C in mixed culture). However, when expressed as a percentage of below ground C allocation, rhizosphere respiration was lower in bromegrass than in clover, respectively, 38% and 45% in monoculture. In mixed culture, this percentage increased by 7.3% for clover, and 3.5% for bromegrass, thus indicating that the interspecific effect of grass was higher than that of clover. The percentage of below-ground C in a soil solution of clover in mixed culture was more than 2-fold that measured in monoculture. It was also significantly correlated with the percentage of below-ground C in respiration. These results provided evidence that the grass-legume mixture has the potential to influence the rhizosphere processes of each species in more than an additive way and that the effect of the interaction was stronger on clover than on bromegrass. The possible implications of this in grass-legume competition are discussed. PMID- 14754917 TI - Effect of K-252a and abscisic acid on the efflux of citrate from soybean roots. AB - The Al-induced release of organic acid has been suggested as an important mechanism for Al resistance in plants. In this study, the effect of K-252a and abscisic acid (ABA) on the efflux of citrate was investigated in soybean (Glycine max L.) roots. Al initiated citrate efflux from the root apices 30 min after the addition of Al. The Al-triggered efflux of citrate was sensitive to metabolic inhibitors and anion channel inhibitors. Pretreatment or treatment with K-252a, an inhibitor of protein kinase, severely inhibited the Al-induced efflux of citrate accompanying an increase in Al accumulation and intensified Al-induced root growth inhibition. Al-treatment increased the endogenous level of abscisic acid (ABA) in soybean roots in a dose- and time-dependent manner, while K-252a failed to inhibit the Al-induced increase in endogenous ABA. Exogenous application of ABA increased the activity of citrate synthase (EC 4.1.3.7) by 26.2%, and decreased Al accumulation by 32.3%, respectively. ABA-induced increases in citrate efflux and root elongation were suppressed by K-252a, while ABA could not reverse the K-252a effects. Taken together, these results suggest that ABA is probably involved in the early response, after which K-252a-sensitive protein kinases play a key step in regulating the activity of an anion channel, through which citrate is released from the apical cells of soybean roots. PMID- 14754918 TI - GFP-labelled Rubisco and aspartate aminotransferase are present in plastid stromules and traffic between plastids. AB - Plastid stromules are membrane-bound protrusions of the plastid envelope that contain soluble stroma. Stromules are often found connecting plastids within a cell and fluorescence recovery after photobleaching (FRAP) experiments have demonstrated that green fluorescent protein (GFP) can move between plastids via these connections. In this report, the ability of endogenous plastid proteins to travel through stromules was investigated. The motility of GFP-labelled plastid aspartate aminotransferase and the Rubisco small subunit was studied in stromules by FRAP. Both fusion proteins assemble into protein complexes that appear to behave similarly to their endogenous counterparts. In addition, both enzymes are capable of trafficking between plastids via stromules. PMID- 14754919 TI - Early responses of resistant and susceptible potato roots during invasion by the potato cyst nematode Globodera rostochiensis. AB - Signals from roots of resistant (cv. Maris Piper) and susceptible (cv. Desiree) potato cultivars during invasion by second stage juveniles (J2s) of the potato cyst nematode, Globodera rostochiensis, were investigated. Novel experimental chambers enabled the recording of electrophysiological responses from roots during nematode invasion. The root cell membrane potentials were maintained throughout the 3 d required to assess invasion and feeding site development. The steady-state resting membrane potentials of Desiree were more negative than those of Maris Piper on day 1, but the reverse on day 3. After 5 d there was no difference between the two cultivars. Intracellular microelectrodes detected marked spike activity in roots after the application of J2s and there were distinct and reproducible differences between the two cultivars, with the response from Desiree being much greater than that from Maris Piper. The responses to mechanical stimulation of roots by blunt micropipettes and sharp electrodes were consistent and similar in both cultivars to the responses in Maris Piper obtained after nematode invasion, but could not account for the marked response found in Desiree. Exogenous application of exoenzymes, used to mimic nematode chemical secretions, resulted in a distinct depolarization pattern that, although similar in both cultivars, was different from patterns obtained during nematode invasion or mechanical stimulation. The pH of homogenates prepared from roots of both cultivars was measured and a Ca2+ channel blocker was used to assess the role of Ca2+ in nematode invasion. The results indicated a role for Ca2+ in the signalling events that occur during nematode invasion. PMID- 14754920 TI - Inhibition of nitrate influx by glutamine in Lolium perenne depends upon the contribution of the HATS to the total influx. AB - Plants of Lolium perenne L. were grown in sterile solution culture supplied with 2 mol m(-3) nitrogen as either nitrate or ammonium. Glutamine at 5 mol m(-3) was added to the nutrient solution of half the plants for 24 h. Root nitrate influx (at external nitrate concentrations 0-2000 mmol m(-3)) and amino acid concentrations were determined. In a second experiment the concentration of the added glutamine was varied from 0-5 mol m(-3) and nitrate influx determined at 250 and 2000 mmol m(-3). The maximum rate of influx attributed to the high affinity transport system (HATS) was reduced by 66% by the presence of glutamine achieved through an 84% reduction in its constitutive component and a 59% reduction in its inducible component. Influx attributed to LATS was unaffected by the addition of glutamine. The inhibition of total nitrate influx by glutamine was positively related to the contribution of HATS to the total influx. In both nitrate- and ammonium-grown plants, the concentration of glutamine required to inhibit nitrate influx significantly was lower when influx was determined at 250 mmol m(-3) compared with 2000 mmol m(-3) nitrate. The addition of glutamine increased its concentrations in root tissue. However, the results cannot be attributed to changes in glutamine alone as its addition also resulted in increased concentrations of other amino acids. Implications for plants growing under field conditions are discussed. PMID- 14754921 TI - Developmental anatomy and auxin response of lateral root formation in Ceratopteris richardii. AB - The homosporous fern Ceratopteris richardii exhibits a homorhizic root system where roots originate from the shoot system. These shoot-borne roots form lateral roots (LRs) that arise from the endodermis adjacent to the xylem poles, which is in contrast to flowering plants where LR formation arises from cell division in the pericycle. A detailed study of the fifth shoot-borne root showed that one lateral root mother cell (LRMC) develops in each two out of three successive merophytes. As a result, LRs emerge alternately in two ranks from opposite positions on a parent root. From LRMC initiation to LR emergence, three developmental stages were identified based on anatomical criteria. The addition of auxins (either indole-3-acetic acid or indole-3-butyric acid) to the growth media did not induce additional LR formation, but exogenous applications of both auxins inhibited parent root growth rate. Application of the polar auxin transport inhibitor N-(1-naphthyl)phthalamic acid (NPA) also inhibited parent root growth without changing the LR initiation pattern. The results suggest that LR formation does not depend on root growth rate per se. The result that exogenous auxins do not promote LR formation in C. richardii is similar to reports for certain species of flowering plants, in which there is an acropetal LR population and the formation of the LRs is insensitive to the application of auxins. It also may indicate that different mechanisms control LR development in non-seed vascular plants compared with angiosperms, taking into consideration the long and independent evolutionary history of the two groups. PMID- 14754922 TI - Effect of salt and osmotic stresses on the expression of genes for the vacuolar H+-pyrophosphatase, H+-ATPase subunit A, and Na+/H+ antiporter from barley. AB - Two cDNA clones encoding vacuolar H+-inorganic pyrophosphatase (HVP1 and HVP10), one clone encoding the catalytic subunit (68 kDa) of vacuolar H+-ATPase (HvVHA A), and one clone encoding vacuolar Na+/H+ antiporter (HvNHX1) were isolated from barley (Hordeum vulgare), a salt-tolerant crop. Salt stress increased the transcript levels of HVP1, HVP10, HvVHA-A, and HvNHX1, and osmotic stress also increased the transcript levels of HVP1 and HvNHX1 in barley roots. The transcription of HVP1 in response to salt stress was regulated differently from that of HVP10. In addition, the HVP1 expression changed in a pattern similar to that of HvNHX1 expression. These results indicate that the expression of HVP1 is co-ordinated with that of HvNHX1 in barley roots in response to salt and osmotic stresses. PMID- 14754923 TI - Isolation and expression pattern of two putative acyl-ACP desaturase cDNAs from Bassia scoparia. AB - The seed lipids of some higher plants contain unusual fatty acids with potentially valuable non-food uses. Seeds of Bassia scoparia contain one such monounsaturated fatty acid, 16:1Delta5. This fatty acid can be used for the production of an insect oviposition pheromone, which is potentially valuable in the control of the mosquito Culex quinquefasciatus, a vector of West Nile virus. Previous work has established that a number of unusual monounsaturated fatty acids are produced by variant forms of the ubiquitous acyl-ACP desaturases. The isolation and initial characterization of two putative acyl-ACP desaturases from B. scoparia, one of which is seed-specific, suggests that such a variant enzyme occurs in this species. PMID- 14754924 TI - ADL2a, like ADL2b, is involved in the control of higher plant mitochondrial morphology. AB - A mitochondrial-GFP construct was used to tag mitochondria fluorescently in a T DNA knockout line for the Arabidopsis dynamin ADL2a. Visualization of mitochondria in vivo demonstrated that disruption of ADL2a affected mitochondrial morphology. Mitochondria in the mutant had a complex morphology; occasionally large spherical organelles could be seen, but, more frequently, the mitochondria adopted a tubular morphology with many constrictions along their length. Mitochondria in the mutant also frequently possessed long protuberances that were named matrixules, extending to many micrometres in length. PMID- 14754926 TI - Chlamydia trachomatis and Chlamydia pneumoniae infections in children and adolescents. PMID- 14754927 TI - Common lower extremity problems in children. PMID- 14754928 TI - Consultation with the specialist: hypospadias. PMID- 14754929 TI - Index of suspicion. PMID- 14754931 TI - Maternal depressive symptoms and adherence to therapy in inner-city children with asthma. AB - CONTEXT: Little is known about how depressive symptoms in mothers affects illness management in inner-city children with asthma. OBJECTIVE: Our goal was to determine how maternal depressive symptoms influence child medication adherence, impact of the child's asthma on the mother, and maternal attitudes and beliefs. METHODS: Baseline and 6-month surveys were administered to 177 mothers of young minority children with asthma in inner-city Baltimore, MD and Washington, DC. Medication adherence, disruptiveness of asthma, and select attitudes toward illness and asthma therapy were measured. Six-month data (N = 158) were used to prospectively evaluate long-term symptom control and emergency department use. Independent variables included asthma morbidity, age, depressive symptoms, and other psychosocial data. RESULTS: No difference in child asthma morbidity was observed between mothers high and low in depressive symptoms. However, mothers with high depressive symptoms reported significantly more problems with their child using inhalers properly (odds ratio [OR]: 5.0; 95% confidence interval [CI]: 1.3-18.9) and forgetting doses (OR: 4.2; 95% CI: 1.4-12.4). Depressive symptoms were also associated with greater emotional distress and interference with daily activities caused by the child's asthma, along with less confidence in asthma medications, ability to control asthma symptoms, and self-efficacy to cope with acute asthma episodes. In addition, depressed mothers reported less understanding about their child's medications and use (OR: 7.7; 95% CI: 1.7 35.9). Baseline asthma morbidity, maternal depression scores, and family income were independently associated with asthma symptoms 6 months later, whereas medication adherence was not predictive of subsequent asthma morbidity or emergency department use. CONCLUSIONS: Maternal depressive symptoms were not associated with child asthma morbidity but were associated with a constellation of beliefs and attitudes that may significantly influence adherence to asthma medications and illness management. Identifying and addressing poor psychological adjustment in mothers is important when developing a child's asthma treatment and may facilitate parent-provider communication, medication adherence, and asthma management among inner-city children. PMID- 14754932 TI - Pediatricians' use of and attitudes about personal digital assistants. AB - BACKGROUND: Personal digital assistants (PDAs) are being increasingly used in medical practice. Although they have been touted as having the ability to improve efficiency and safety, little is known about pediatricians' use of and attitudes about PDAs. OBJECTIVE: Our goals were to 1) determine the percentage of pediatricians using PDAs and computers, 2) determine perceived strengths and weaknesses of PDAs, and 3) explore characteristics associated with beliefs and use. DESIGN/METHODS: Pediatricians (2130) were selected randomly from the American Medical Association Physician Masterfile of US-licensed physicians. All participants were mailed a survey along with a prepaid return envelope and a 1 dollar incentive. Up to 3 mailings were sent per participant. RESULTS: Of eligible participants, 63.2% returned a survey. There were no significant differences between respondents and nonrespondents with respect to sex, type of practice, and present employment. Thirty-five percent of respondents currently use PDAs at work, and 40% currently use PDAs for personal use. Of those using PDAs, the most commonly used applications were for drug reference (80%), personal scheduling (67%), and medical calculations (61%). Few pediatricians are currently using PDAs for prescription writing (8%) or billing (4%). Users of PDAs were more likely to be male (adjusted odds ratio [AOR]: 2.29; 95% confidence interval [CI]: 1.64-3.19), in an urban community (AOR: 1.81; 95% CI: 1.30-2.55), in training (AOR: 2.64; 95% CI: 1.58-4.42), not in private practice (AOR: 1.47; 95% CI: 1.03 2.11), and a more recent graduate of medical school (AOR: 1.04 per year; 95% CI: 1.02-1.06). When controlling for covariates, those using PDAs were more likely to believe that PDAs can decrease medical errors (AOR: 2.22; 95% CI: 1.46-3.38) and increase efficiency (AOR: 2.40; 95% CI: 1.56-3.71). When compared with nonusers, users were less likely to view the small screen size (AOR: 0.53; 95% CI: 0.37 0.77) or system speed (AOR: 0.47; 95% CI: 0.26-0.84) as a problem but were significantly more likely to view memory as an issue (AOR: 3.48; 95% CI: 2.30 5.25). CONCLUSIONS: More than one third of pediatricians are using PDAs in clinical practice. There seems to be a general consensus among users that they have the potential to improve patient safety and streamline care. Future studies should explore means to utilize their potential. PMID- 14754933 TI - A bitter pill: attempting change in a pediatric morning report. AB - OBJECTIVE: To assess and address participants' dissatisfaction with departmental morning report (MR). METHODS: Three consecutive MR sessions were observed, and those data, in combination with findings from the literature, were used to guide creation of a quantitative survey. The survey was administered to all faculty and housestaff. Survey items addressed the educational focus, leadership, and format of MR and the value of specific educational conferences. Subsequently, 2 interventions were developed to increase participants' level of satisfaction with MR. The interventions' effect was measured 1 year later using a second survey. RESULTS: Eighty-two percent of housestaff and 43% of faculty responded to the first survey. Our findings are contrary to those in the literature. For example, respondents rated the presence of all faculty at MR as desirable, and both subspecialty and general pediatric contributions were considered important. Housestaff assigned greater educational value to all teaching conferences than did faculty. Similarly, housestaff believed more strongly than faculty that MR leadership should be changed. A follow-up survey demonstrated a decrease in mean perceived value of MR after our interventions. CONCLUSIONS: Despite aggressive research, we failed to identify the source of participants' dissatisfaction. Although our interventions were based on research data and reflected participants' perceptions and concerns related to MR, they were not supported by the faculty. This demonstrates the inherent difficulties in changing an "institution" such as MR. Thus, examination of MR goals and satisfaction by individual training programs should be conducted within the confines of the conference's preexisting structure, without attempt to apply literature-driven expectations. PMID- 14754934 TI - Transtelephonic electrocardiographic monitors for evaluation of children and adolescents with suspected arrhythmias. AB - OBJECTIVE: Patient-activated transtelephonic electrocardiographic event monitors (TTMs) are often used for the evaluation of children and adolescents with suspected arrhythmias. Since their introduction 25 years ago, there has been little inquiry quantifying the usefulness of TTMs for pediatric patients. The objective of this study was to measure the utility of TTMs for children and adolescents with symptoms of a possible cardiac rhythm disturbance. METHODS: Medical records of all patients who received TTMs from C.S. Mott Children's Hospital Electrocardiography Laboratory between February 1, 1993, and October 31, 2000, were reviewed. Patients with pacemakers, known arrhythmias, or age older than 18 years were excluded. Indications for monitoring included palpitations with or without other symptoms (N = 420), chest discomfort (N = 43), and presyncope or syncope (N = 32). RESULTS: A total of 495 studies (patient mean age: 10.2 +/- 4.3 years; range: 0.1-17.9 years; 48% male) met inclusion criteria. Monitoring was performed for 1 to 1021 consecutive days (mean: 103 +/- 97). Fifty two percent (N = 257) of patients failed to transmit an electrocardiogram while experiencing symptoms. Fewer boys transmitted electrocardiograms (N = 100/238). Of 238 symptomatic patients, 15% (N = 35; mean age: 11.4 +/- 4.7 years; range: 0.1-17.4 years; 51% male) had supraventricular tachycardia (SVT). No other significant arrhythmia that may warrant treatment was identified. All patients with SVT had palpitations. No patients with isolated chest discomfort, presyncope, or syncope had SVT (N = 75). SVT was documented more frequently in patients with postevent (N = 35/464) than loop recorders (N = 0/31). Of those with SVT, 71% (N = 25) and 91% (N = 33) transmitted events within 4 and 16 weeks, respectively. Follow-up for 1 to 108 months (mean: 32 +/- 25; median: 26) in 53% (243 of 460) of patients without SVT uncovered a 3% (N = 7) rate of subsequent SVT detection. The overall sensitivity of the TTM test was 83% (35 of 42) for detection of SVT. The sensitivity of studies theoretically limited to 4 and 16 weeks would be 60% (25 of 42) and 79% (33 of 42), respectively. The negative predictive value of the TTM study was 99% in our patient population. The negative predictive value of tests theoretically limited to 4 and 16 weeks would be 96% and 98%, respectively. TTM studies of 2 weeks' duration were most cost-effective in terms of total diagnostic yield. In contrast, studies of 4 weeks' duration were most cost-efficient for SVT detection. CONCLUSIONS: TTMs are useful for the evaluation of children and adolescents with palpitations but not with isolated chest pain, syncope, or presyncope. In this study, girls were more likely to transmit events. The sensitivity of TTMs for detection of SVT was 83%. The negative predictive value of the TTM test was 99%. Monitoring for longer than 16 weeks did not increase test sensitivity. Studies of 4 weeks' duration proved most cost-effective for SVT detection. PMID- 14754935 TI - Educating the pediatrician of the 21st century: defining and implementing a competency-based system. AB - PURPOSE: The final product of the Accreditation Council for Graduate Medical Education (ACGME) Outcomes Project is to improve health care through ensuring the competence of physicians during their training and beyond. Its success depends on a paradigm shift to a competency-based model of medical education. This article presents an overview of the model and describes the first stages of this multistep transition in a university setting. Our hope is to raise awareness of the impact on the pediatric community at large and provide a foundation on which other educators can continue to build. METHODS: The ACGME established 6 domains of competence as the first step in the paradigm shift. To begin work on the second step of establishing benchmarks for evaluation of competence and thresholds at which they should be achieved, the educational leadership from the primary care departments formed a work group with the support of a Health Resources and Services Administration grant. RESULTS: Review of the literature led us to embark on a stepwise process for initiating a competency-based system of medical education. We established the benchmarks by a process of individual effort followed by group consensus. These benchmarks, in the aggregate, permit an evaluator to determine whether a given competency has been achieved. The next phases of step 2 required setting thresholds for the demonstration of competence and finally establishing consensus regarding the threshold for competence at each level of training for each specific benchmark. To accomplish the latter, we surveyed the program directors of the 202 accredited pediatric training programs that are members of the Association of Pediatric Program Directors. Eighty-one members (40%) completed our survey. Although some controversy existed for thresholds during the postgraduate-year-1 level, majority consensus was reached for nearly all benchmarks at the upper levels of training. Impact of the shift to competencies for medical students and practicing physicians is also reviewed. CONCLUSION: Having defined the benchmarks and thresholds for the 6 ACGME domains of competence, we are well positioned to move to step 3: developing the necessary tools to evaluate competence. The move to competency-based education impacts the entire pediatric community. Lessons learned from our experience may be generalized and thus be of value to the community at large. PMID- 14754936 TI - Age at first measles-mumps-rubella vaccination in children with autism and school matched control subjects: a population-based study in metropolitan atlanta. AB - OBJECTIVE: To compare ages at first measles-mumps-rubella (MMR) vaccination between children with autism and children who did not have autism in the total population and in selected subgroups, including children with regression in development. METHODS: A case-control study was conducted in metropolitan Atlanta. Case children (N = 624) were identified from multiple sources and matched to control children (N = 1824) on age, gender, and school. Vaccination data were abstracted from immunization forms required for school entry. Records of children who were born in Georgia were linked to Georgia birth certificates for information on maternal and birth factors. Conditional logistic regression was used to estimate odds ratios (ORs). RESULTS: The overall distribution of ages at MMR vaccination among children with autism was similar to that of matched control children; most case (70.5%) and control children (67.5%) were vaccinated between 12 and 17 months of age. Similar proportions of case and control children had been vaccinated before 18 or before 24 months. No significant associations for either of these age cutoffs were found for specific case subgroups, including those with evidence of developmental regression. More case (93.4%) than control children (90.6%) were vaccinated before 36 months (OR: 1.49; 95% confidence interval: 1.04-2.14 in the total sample; OR: 1.23; 95% confidence interval: 0.64 2.36 in the birth certificate sample). This association was strongest in the 3- to 5-year age group. CONCLUSIONS: Similar proportions of case and control children were vaccinated by the recommended age or shortly after (ie, before 18 months) and before the age by which atypical development is usually recognized in children with autism (ie, 24 months). Vaccination before 36 months was more common among case children than control children, especially among children 3 to 5 years of age, likely reflecting immunization requirements for enrollment in early intervention programs. PMID- 14754937 TI - Perception of child vulnerability among mothers of former premature infants. AB - OBJECTIVES: Parents of premature infants often perceive their infants as medically vulnerable. High parental perception of child vulnerability (PPCV) is associated with disproportionately high health care utilization. The objectives of this study were to determine whether higher PPCV is correlated with worse developmental outcome in premature infants at 1-year adjusted age and to identify factors, present at neonatal discharge, that predict high PPCV. METHODS: This prospective cohort study assessed mothers of 116 premature infants who were or = 18 years (mean 56 years) who attended a primary care clinic in San Diego, California in 1995-1997. MAIN OUTCOME MEASURE: Adolescent pregnancy, psychosocial consequences, and fetal death, compared by ACE score (emotional, physical, or sexual abuse; exposure to domestic violence, substance abusing, mentally ill, or criminal household member; or separated/divorced parent). RESULTS: Sixty-six percent (n = 6015) of women reported > or = 1 ACE. Teen pregnancy occurred in 16%, 21%, 26%, 29%, 32%, 40%, 43%, and 53% of those with 0, 1, 2, 3, 4, 5, 6, and 7 to 8 ACEs. As the ACE score rose from zero to 1 to 2, 3 to 4, and > or = 5, odds ratios for each adult consequence increased (family problems: 1.0, 1.5, 2.2, 3.3; financial problems: 1.0, 1.6, 2.3, 2.4; job problems: 1.0, 1.4, 2.3, 2.9; high stress: 1.0, 1.4, 1.9, 2.2; and uncontrollable anger: 1.0, 1.6, 2.8, 4.5, respectively). Adolescent pregnancy was not associated with any of these adult outcomes in the absence of childhood adversity (ACEs: 0). The ACE score was associated with increased fetal death after first pregnancy (odds ratios for 0, 1-2, 3-4, and 5-8 ACEs: 1.0, 1.2, 1.4, and 1.8, respectively); teen pregnancy was not related to fetal death. CONCLUSIONS: The relationship between ACEs and adolescent pregnancy is strong and graded. Moreover, the negative psychosocial sequelae and fetal deaths commonly attributed to adolescent pregnancy seem to result from underlying ACEs rather than adolescent pregnancy per se. PMID- 14754945 TI - The effects of obesity, gender, and ethnic group on left ventricular hypertrophy and geometry in hypertensive children: a collaborative study of the International Pediatric Hypertension Association. AB - OBJECTIVE: To determine the prevalence of left ventricular hypertrophy (LVH) in a multiethnic group of children and adolescents with hypertension. DESIGN/METHODS: Pooled data from 1998 to 2001 from 3 sites belonging to the International Pediatric Hypertension Association were reviewed. Patients undergoing echocardiography to detect LVH as part of the evaluation for hypertension were included for analysis. Left ventricular mass was calculated from 2-dimensional guided M-mode echocardiographic measurements of the left ventricle. Left ventricular mass index (LVMI) was calculated as left ventricular mass/height(2.7). LVH by adult criteria was defined as LVMI > 51 g/m(2.7) and by pediatric criteria as LVMI > 38.6 g/m(2.7). Left ventricle geometry was classified as concentric, concentric remodeling, eccentric, or normal. RESULTS: Data on 129 patients with a mean age of 13.6 +/- 3.6 years were analyzed. The population was 67% male, 46.5% white, 38.0% African American, and 15.5% Hispanic. The prevalence of LVH was 15.5% using adult criteria and 41.1% using pediatric criteria. Increasing body mass index (BMI) was associated with a higher LVMI. Using either pediatric or adult criteria LVH was associated with BMI > or =95th percentile for age and gender. LVH and concentric hypertrophy were identified most frequently in Hispanic children. CONCLUSIONS: LVH occurs commonly in children with hypertension and is associated with an increased BMI. LVH may be more prevalent in Hispanic children than in other ethnic groups. Prevention and treatment of obesity is important in reducing the cardiovascular risk for children with hypertension. Further evaluation of the frequency of LVH in multiethnic populations is needed. PMID- 14754946 TI - Coping, commitment, and attitude: quantifying the everyday burden of enuresis on children and their families. AB - OBJECTIVE: To develop and evaluate a parent-completed questionnaire for use by clinicians as part of routine care to assess the burden of diurnal and nocturnal enuresis on children and their families. METHODS: The questionnaire consisted of items that measure the impact on the child and his/her parent, the child's coping ability and commitment to treatment, previous treatment success, family frustration and overall cohesion, and parental attitudes about enuresis and its treatment. Questionnaires (n = 208) were completed by parents during the child's scheduled office visit for enuresis at 5 specialty clinics across the United States. Traditional criteria were used to assess reliability and validity of the questionnaire, including analysis of variance. RESULTS: Success rates provide evidence that many of the items in the child scale (79%) and all items in the parent scale (100%) met stringent criteria. alpha values were.62 and.77, respectively. Statistically significant differences were observed for the scales across responses on all but 1 global item, the majority of parental attitude items, whether the child urinated at bedtime, and the number of pads used. These findings suggest that the child's coping ability and commitment and the family's overall cohesion and frustration with the problem influence parental perceptions about the impact of enuresis on the child and the family. CONCLUSIONS: Findings about the performance of the new measure were satisfactory and suggest that, after further refinement, it should prove as a useful tool for clinicians treating enuresis in children. PMID- 14754948 TI - Adenotonsillectomy in children with obstructive sleep apnea syndrome reduces health care utilization. AB - OBJECTIVE: To investigate health care utilization of children with obstructive sleep apnea syndrome (OSAS) 1 year after adenotonsillectomy (T&A). METHODS: A longitudinal, case-controlled, prospective study was conducted at Clalit Health Care Services (CHS), a health maintenance organization in the southern region of Israel. We defined 3 groups of children: 1) children who had OSAS and were treated with T&A (n = 130); 2) children who had OSAS and did not undergo surgery (n = 90); and 3) control subjects who were matched by age, sex, and area of residency (n = 520) and randomly selected from the CHS database. OSAS was verified with polysomnography studies in all patients. Indices of health care utilization were analyzed 1 year before and 1 year after T&A. Medical records in the emergency department and during hospitalization were reviewed for diagnosis before the polysomnography diagnosis. RESULTS: Mean age of all children with OSAS was 5.6 +/- 3.6 years. Total annual health care costs were reduced by one third in children who underwent T&A, whereas there was no change in the control and untreated OSAS groups. T&A was associated with a 60% reduction in the number of new admissions, 39% reduction in emergency department visits, 47% reduction in the number of consultations, and 22% reduction in costs for prescribed drugs. In group 2, the total costs were similar in years 1 and 2. CONCLUSIONS: T&A significantly reduces health care utilization in children with OSAS. Untreated children with moderate and severe OSAS will continue to consume high levels of health care resources. Increased morbidity among children with OSAS is mainly related to upper respiratory tract infections. PMID- 14754947 TI - Does environment mediate earlier onset of the persistent childhood asthma phenotype? AB - OBJECTIVE: We investigated the role of environmental and hereditary factors in determining whether persistent childhood wheezing phenotypes had an early or late onset. METHODS: In a whole population birth cohort (n = 1456), children were seen at birth and at 1, 2, 4, and 10 years. At each visit, information was collected prospectively regarding wheeze prevalence and used to classify subjects into wheezing phenotypes. Information on genetic and environmental risk factors in early life was also obtained prospectively, and skin-prick testing to common allergens was performed at 4 years. RESULTS: Early-onset persistent wheezers (n = 125) had wheeze onset in the first 4 years, still present at age 10, whereas late onset persistent wheezers (n = 81) had wheeze onset after age 4 years that was still present at 10 years. Multivariate logistic regression analysis identified independent significance only for inherited factors (parental asthma, family history of rhinitis, eczema at 4 years, and atopic status at 4 years) in the development of late-onset persistent wheeze. However, low social class at birth, recurrent chest infections at 2 years, and parental smoking at 2 years plus inherited factors (eczema at 2 years; food allergy at 4 years; maternal asthma, sibling asthma, maternal urticaria, and atopic status at 4 years) demonstrated independent significance for early-onset persistent wheeze. CONCLUSION: Inheritance seems to be of prime significance in the cause of persistent childhood wheeze. Environmental exposure in early life may combine with this tendency to produce an early onset of persistent wheeze. Absence of these environmental factors might delay but not prevent the onset of wheeze in children with atopic heredity. PMID- 14754949 TI - Abnormal auditory brainstem response among infants with prenatal cocaine exposure. AB - OBJECTIVE: To determine the auditory brainstem response (ABR) patterns in infants with prenatal cocaine exposure as detected by meconium analysis. METHODS: Meconium drug analysis was done on term infants who were at risk for antenatal illicit drug exposure. Infants who tested positive for cocaine served as the exposed group; those who tested negative served as control subjects. An ABR was done on the infants within 6 days of life. Absolute latencies at 40 and 70 decibels (dB) of waves I, III, and V and interpeak latencies between I and III, III and V, and I and V were measured. RESULTS: Among 58 infants in the study, 21 (35.6%) were positive, by meconium analysis, for cocaine and 5 (8.5%) were positive for cannabinoids. Four infants tested positive for cocaine, although their mothers denied illicit drug use. There was a significant difference in mean maternal age, gravidity, parity, birth weight, and head circumference among cocaine-exposed versus nonexposed infants. The ABR from cocaine-positive infants showed significantly prolonged mean absolute latencies in both ears as compared with nonexposed infants: right ear at 40 dB III (4.823 vs 4.447 milliseconds), 40 dB V (7.400 vs 6.968 milliseconds), 70 dB I (2.111 vs 1.631 milliseconds), and 70 dB III (4.122 vs 3.821 milliseconds); left ear at 40 dB III (4.820 vs 4.444 milliseconds), 40 dB V (7.460 vs 6.860 milliseconds), 70 dB I (2.063 vs 1.741 milliseconds), 70 dB III (4.026 vs 3.656 milliseconds), and 70 dB V (6.568 vs 6.258 milliseconds). The interpeak latency was not significantly different except in 1 cocaine-positive infant: left ear at 40 dB III to V (2.667 vs 2.417 milliseconds). CONCLUSION: The ABR in neonates who are exposed prenatally to cocaine shows prolonged absolute peak latencies compared with nonexposed neonates and may indicate compromise of the auditory system from gestational exposure to cocaine that will need additional audiologic follow-up. Meconium analysis can more accurately detect the infants who are at risk. PMID- 14754950 TI - Ultrasound imaging of milk ejection in the breast of lactating women. AB - OBJECTIVE: Currently, the methods for assessing milk ejection in women include serial sampling of plasma oxytocin and measurement of intraductal pressure, both of which are invasive and may induce stress. We hypothesized that milk ejection would cause an increase in milk-duct diameter that could be observed noninvasively with ultrasound, and this could be used to investigate the physiology of milk ejection in women. METHODS: One milk duct was scanned in the unsuckled breast in 2 groups of mothers: group BB (n = 21) for the beginning of a breastfeed and group EB (n = 24) for the entire breastfeed. A duct also was monitored for a 5-minute period on 2 separate days in the absence of factors that may induce milk ejection in group EB to provide a baseline duct diameter. Milk intake at a breastfeed was measured by test weighing. RESULTS: A significant increase in milk-duct diameter was observed when milk ejection was sensed and/or the infant changed its swallowing pattern in both groups. Multiple increases and decreases (mean: 2.5 per breastfeed; standard deviation: 1.5; n = 62) in duct diameter occurred in group EB. Duct diameter remained relatively stable between breastfeeds (coefficient of variation: 1.4%-8.3%). Infant milk intake was positively related to the number of milk ejections (r2 =.365; n = 57). CONCLUSIONS: Ultrasound is an objective, noninvasive technique for detecting milk ejection by observing an increase in milk-duct diameter. However, this technique requires an experienced ultrasonographer, adequate imaging time, and surroundings conducive to breastfeeding. Multiple milk ejections were common during breastfeeding, although they were not sensed by mothers. The number of milk ejections influenced the amount of milk the infant consumed. PMID- 14754951 TI - Maternal selective serotonin reuptake inhibitor use during pregnancy and newborn neurobehavior. AB - OBJECTIVE: This is a prospective study of the effects of maternal use of selective serotonin reuptake inhibitors (SSRIs) during pregnancy on newborn neurobehavioral integrity, including systematic measures of behavioral state, sleep organization, motor activity, heart rate variability (HRV), tremulousness, and startles. METHODS: The sample included 17 SSRI-exposed and 17 nonexposed, full-birth-weight newborn infants who had no obvious medical problems and were matched on maternal cigarette use, social class, and maternal age. SSRI exposure was determined by medical records and maternal self-report during a standard interview. Behavioral state, startles, and tremulousness were evaluated for 1 hour between feedings. Automated recordings of motor activity and HRV were also assessed during a 15-minute subset sleep period. HRV was subjected to spectral analysis to detect rhythms in autonomic regulation. Exposed and nonexposed infant groups were compared on measures of neurobehavioral development both before and after adjustment for gestational age as a covariate. RESULTS: SSRI-exposed infants had a shorter mean gestational age; were more motorically active and tremulous; and showed fewer rhythms in HRV, fewer changes in behavioral state, fewer different behavioral states, and a lower peak behavioral state. SSRI exposed infants also had significantly more rapid eye movement sleep, which was characterized by longer continuous bouts in that state and higher numbers of spontaneous startles or sudden arousals. After effects of gestational age were covaried, significant differences continued to be found in tremulousness and all measures of state and sleep organization, but effects on startles, motor activity, and rhythms in HRV were no longer significant. CONCLUSIONS: Results provide the first systematic evidence that women who use SSRIs during pregnancy have healthy, full-birth-weight newborn infants who show disruptions in a wide range of neurobehavioral outcomes. Effects on motor activity, startles, and HRV may be mediated through the effects of SSRI exposure on gestational age. Future research can lead to a better understanding of the effects of SSRI use during pregnancy and an improved public health outcome. PMID- 14754952 TI - Radical reduction in the rate of extensive corrective surgery for clubfoot using the Ponseti method. AB - OBJECTIVES: The purpose of this study was to evaluate the efficacy of the Ponseti method in reducing extensive corrective surgery rates for congenital idiopathic clubfoot. METHODS: Consecutive case series were conducted from January 1991 through December 2001. A total of 157 patients (256 clubfeet) were evaluated. All patients were treated by serial manipulation and casting as described by Ponseti. Main outcome measures included initial correction of the deformity, extensive corrective surgery rate, and relapses. RESULTS: Clubfoot correction was obtained in all but 3 patients (98%). Ninety percent of patients required or =70). CP was identified in 35 of 366 3-year-olds (9.5%). An FSIQ <70 was identified in 47 of 366 children at 8 years old (12.8%). FSIQ <70 occurred in 14 of 17 children with tri- or quadriplegia (82%), 8 of 18 children with di- or hemiplegia (44%), and 25 of 331 children without CP (7.5%). Useful LRs were calculated for tri- or quadriplegia (30), di- or hemiplegia (5.7), and children without CP (0.55). These LRs have greater impact on posttest odds for FSIQ <70 than those for birth weight <1000 g, history of bronchopulmonary dysplasia, and Stanford-Binet Intelligence Score <70 at age 3. We conclude that the neurologic examination at 3 years old predicts FSIQ <70 at age 8 with LRs that allow evidence-based parental counseling and intervention planning. PMID- 14754963 TI - Statement of endorsement. Pediatric care in the emergency department. PMID- 14754964 TI - Risperidone-associated diabetes mellitus in children. PMID- 14754965 TI - Management of positional skull deformities: who needs a helmet? PMID- 14754966 TI - Maternal depression and the pediatrician. PMID- 14754967 TI - Body mass index graphs for children. PMID- 14754968 TI - Uninsured children: how we count matters. PMID- 14754969 TI - Family pediatrics. PMID- 14754970 TI - 2001 American Academy of Pediatrics practice parameter on attention deficit/hyperactivity disorder. PMID- 14754971 TI - Infants with bilirubin levels of 30 mg/dL or more. PMID- 14754972 TI - Interpreting epidemiologic research and childhood leukemia. PMID- 14754973 TI - Management of hypoplastic left heart syndrome. PMID- 14754974 TI - Fillers. PMID- 14754975 TI - Abnormal retinal vascular morphology in young adults following intrauterine growth restriction. AB - OBJECTIVE: Intrauterine growth restriction (IUGR) resulting in low birth weight for gestational age may predispose one to development of cardiovascular disease later in life. Abnormal fetal blood flow in the presence of fetal growth restriction helps to distinguish infants with true fetal growth impairment from small but normal infants. Our goal was to investigate associations between IUGR with abnormal fetal blood flow and abnormal retinal vascular morphology at 18 years of age. METHODS: A prospective study was performed with 21 subjects with IUGR (abnormal fetal aortic blood flow velocity; birth weight small for gestational age; median birth weight deviation from the population mean of -31% [range: -22% to -42%] and in 23 subjects with birth weight appropriate for gestational age [normal fetal aortic blood flow velocity; median birth weight deviation of -2% (range: -10% to 22%)]). The retinal vessel morphology was evaluated by digital image analysis. RESULT: Subjects with IUGR (n = 21) had significantly less retinal vascularization as evidenced by a lower number of vascular branching points (median: 26; range: 20-31) as compared with the subjects who were born appropriate for gestational age (median: 28; range: 26 32). Within the entire group (N = 44), increasing negative birth weight deviation was associated with a reduced number of vascular branching points (r = 0.36. CONCLUSION: Our findings show that IUGR with abnormal fetal blood flow is associated with abnormal retinal vascular morphology in young adult life. PMID- 14754976 TI - Does breastfeeding protect against pediatric overweight? Analysis of longitudinal data from the Centers for Disease Control and Prevention Pediatric Nutrition Surveillance System. AB - OBJECTIVE: To examine whether increasing duration of breastfeeding is associated with a lower risk of overweight in a low-income population of 4-year-olds in the United States. METHODS: Visit data were linked to determine prospectively the duration of breastfeeding (up to 2 years of age) and weight status at 4 years of age. Overweight among 4-year-old children was defined as a body mass index (BMI) for-age at or above the 95th percentile based on the 2000 Centers for Disease Control and Prevention growth charts. Logistic regression was performed, controlling for gender, race/ethnicity, and birth weight. In a subset of states, links to maternal pregnancy records also permitted regression analysis controlling for mother's age, education, prepregnancy BMI, weight gain during pregnancy, and postpartum smoking. Data from the Pediatric Nutrition Surveillance System, which extracts breastfeeding, height, and weight data from child visits to public health programs, were analyzed. In 7 states, data were linked to Pregnancy Nutrition Surveillance System data. A total of 177 304 children up to 60 months of age were included in our final pediatric-only analysis, and 12 587 were included in the pregnancy-pediatric linked analysis. RESULTS: The duration of breastfeeding showed a dose-response, protective relationship with the risk of overweight only among non-Hispanic whites; no significant association was found among non-Hispanic blacks or Hispanics. Among non-Hispanic whites, the adjusted odds ratio of overweight by breastfeeding for 6 to 12 months versus never breastfeeding was 0.70 (95% confidence interval: 0.50-0.99) and for >12 months versus never was 0.49 (95% confidence interval: 0.25-0.95). Breastfeeding for any duration was also protective against underweight (BMI-for-age below the 5th percentile). CONCLUSION: Prolonged breastfeeding is associated with a reduced risk of overweight among non-Hispanic white children. Breastfeeding longer than 6 months provides health benefits to children well beyond the period of breastfeeding. PMID- 14754977 TI - Twelve-month safety and efficacy of inhaled fluticasone propionate in children aged 1 to 3 years with recurrent wheezing. AB - OBJECTIVE: Our aim was to compare the 12-month safety and efficacy of fluticasone propionate (FP) and sodium cromoglycate (SCG) in children aged 1 to 3 years with mild to moderate recurrent wheeze. METHODS: The study was a randomized, parallel group, open-label multicenter study of 625 children, aged 1 to 3 years, with recurrent wheeze randomized in a 3:1 ratio to treatment for 52 weeks with FP (100 microg twice daily) via metered-dose inhaler and Babyhaler spacer device or SCG (5 mg 4 times daily) via metered-dose inhaler and Nebuhaler spacer device, respectively. RESULTS: There was no significant difference in mean adjusted growth rates between the 2 groups: 84.0 mm/year in the FP group versus 86.4 mm/year in the SCG group (difference FP-SCG: -2.4 mm/year; 95% confidence interval: -6.6 to 1.8). Growth comparisons were independent of age, gender, previous use of steroid, or whether measured as length and/or height. Serum and urinary cortisol concentrations showed a statistically significant suppression of 10% and 14%, respectively, but the number of patients with serum cortisol levels below the lower normal limit was reduced during the trial. Both treatments were well tolerated. The most common drug-related adverse events were cough (2% FP vs 1% SCG) and hoarseness (1% FP vs 0% SCG). One incident of cataract was observed at baseline and 1 after FP treatment; the latter had resolved after 12 months. The efficacy of FP was superior to SCG with fewer cases of symptom worsening, exacerbations, and requirements for oral steroid treatment and more symptom-free days and days without use of rescue treatment. CONCLUSIONS: Twelve months of treatment with inhaled FP (100 microg twice daily) in preschool children aged 1 to 3 years with recurrent wheeze has no effect on growth and no other clinically important side effects but is more efficacious than SCG. PMID- 14754978 TI - Cost-utility analysis of orthoptic screening in kindergarten: a Markov model based on data from Germany. AB - OBJECTIVE: To estimate the long-term cost-effectiveness of a hypothetical screening program for untreated amblyopia in 3-year-old children conducted by orthoptists in all German kindergartens in the year 2000. METHODS: A cost-utility analysis was performed for which a decision tree was combined with a Markov model. Incremental costs and effects during the children's remaining lifetime were estimated. The model took into account the probability of treatment without screening, age-specific treatment success rates, costs of screening and treatment, as well as effects of unilateral and bilateral visual impairment caused by amblyopia and other eye diseases coming along later in life on quality of life (utility). Model parameter values were obtained from a field study of orthoptic screening in kindergarten, from the literature, and from expert interviews. Costs were estimated from a third-party payer perspective. Uncertainty was assessed by univariate and probabilistic sensitivity analysis (Monte Carlo simulation). RESULTS: The incremental cost-effectiveness ratio (ICER) of orthoptic screening was 7397 Euro (euro) per quality-adjusted life year (QALY) when costs and effects were discounted at 5%. In univariate sensitivity analysis, the ICER was sensitive to the uncertainty regarding the utility of unilateral visual impairment and to the discount rate for effects; besides uncertainty regarding the prevalence of untreated amblyopia, the odds ratio of success of treatment when started late, and the probability of treatment without screening had a noticeable but much smaller effect. Monte Carlo simulation yielded a 90% uncertainty interval for the ICER of 3452 euro/QALY to 72 637 euro/QALY; the probability of an ICER <25 000 euro/QALY was 84%. CONCLUSIONS: The ICER of orthoptic screening seems to fall within a range that warrants careful consideration by decision-makers. Much of the uncertainty in results comes from the uncertainty regarding the effect of amblyopia on quality of life. To reduce this uncertainty, the impact of amblyopia on utility should be investigated. PMID- 14754979 TI - Unmet need for routine and specialty care: data from the National Survey of Children With Special Health Care Needs. AB - OBJECTIVE: To assess the prevalence of unmet needs for routine and specialty care among children with special health care needs (CSHCN) and to identify factors associated with the likelihood of having unmet need for medical care. METHODS: Data come from the respondents for 38 866 children interviewed for the National Survey of Children With Special Health Care Needs. Bivariate analyses were used to assess differences in unmet need for medical care by various environmental, predisposing, enabling, and need factors. Logit analyses were used to determine independent effects of these variables on the likelihood of having an unmet need for medical care. RESULTS: Nationally, 74.4% and 51.0% of CSHCN needed routine and subspecialty physician care, respectively. Of those reporting that they needed routine care, 3.2% were unable to obtain these services. Of those reporting a need for specialty care, 7.2% reported not obtaining all needed specialty care. The prevalence of unmet need for specialty care significantly exceeded the prevalence of unmet need for routine care. In logit analyses, African American children and children whose mothers had less than a high-school education faced twice the odds of having an unmet need for routine care. Compared with nonpoor children, children living below the federal poverty level were significantly more likely to have an unmet need for routine (adjusted odds ratio [aOR]: 1.97; 95% confidence interval [CI]: 1.23-3.14) and specialty (aOR: 2.50; 95% CI: 1.49-4.18) care. Near-poor children were also significantly more likely than nonpoor children to have unmet needs for routine and specialty care. Uninsured children were significantly more likely than continuously insured children to report an unmet need for routine (aOR: 7.51; 95% CI: 4.99-11.30) and specialty (aOR: 4.29; 95% CI: 2.99-6.15) care. Our findings also show that higher levels of general pediatrician supply, relative to the pediatric population, are associated with a significantly lower likelihood of having an unmet need for routine care. Likewise, a greater supply of pediatric subspecialists is associated with a decreased likelihood of having an unmet need for specialty care. CONCLUSIONS: Compared with previous reports of the general pediatric population, CSHCN have higher levels of unmet need for medical services. Our regression results emphasize that children vulnerable because of their social circumstances (eg, poverty, etc) have significantly greater odds of having unmet need for routine and specialty physician care. Furthermore, our findings highlight the importance of insurance coverage in ensuring access to needed routine and specialty medical services. PMID- 14754980 TI - Central nervous system side effects of first- and second-generation antihistamines in school children with perennial allergic rhinitis: a randomized, double-blind, placebo-controlled comparative study. AB - OBJECTIVE: Allergic rhinitis is common and on the rise. Antihistamines are the mainstay of treatment and are the most commonly prescribed drugs in Singapore. Treatment-related sedation and its effect on cognition are a major concern. First and second-generation antihistamines show varying degrees of sedation, but to date, objective studies in children are lacking. The objective of this study was to assess the sedating effect of cetirizine (second-generation antihistamine) and chlorpheniramine (first-generation antihistamine) compared with placebo using an objective neurophysiological test. METHODS: This was a prospective, double-blind, placebo-controlled, randomized, single-dose, 3-way crossover study. Twenty-four children aged 7 to 14 years with allergic rhinitis completed the study. All children were randomly allocated to medication sequences and received 3 different drugs on 3 different days, at least 1 week apart. The P300 event-related potential was used as an objective test of sedation. Subjective assessment was by a visual analog scale. RESULTS: Chlorpheniramine and cetirizine increased P300 latency when compared with baseline. No significant increase was obtained with placebo. The significant increase in P300 latency was not accompanied by significant change in subjective somnolence as measured by the visual analog scale. CONCLUSION: We have shown that cetirizine has sedative properties in children. The lack of correlation between P300 latency and the visual analog scale indicates that sedation induced by these drugs may not be subjectively noted. PMID- 14754981 TI - Nonmyeloablative hematopoietic stem cell transplant for X-linked hyper immunoglobulin m syndrome with cholangiopathy. AB - OBJECTIVE: X-linked hyper-immunoglobulin M (X-HIM) syndrome is a rare genetic immunodeficiency syndrome caused by mutations in the gene encoding CD40 ligand (CD40L, CD154). Allogeneic hematopoietic stem cell transplantation (HSCT) offers the prospect of immune reconstitution in X-HIM syndrome. Standard HSCT using high dose chemoradiotherapy can be followed by serious hepatic problems, including veno-occlusive disease, graft-versus-host disease, and/or drug-induced hepatotoxicity. In patients whose liver function is compromised before HSCT, such as in X-HIM syndrome caused by cholangiopathy and hepatitis related to opportunistic infections, there is a higher likelihood of hepatotoxicity. We explored nonmyeloablative HSCT in 2 patients with X-HIM syndrome. Nonmyeloablative HSCT without liver transplant for X-HIM syndrome, to our knowledge, has not been described previously. METHODS: Two children with X-HIM syndrome and persistent infections had documented cholangiopathy on liver biopsy. Both children underwent nonmyeloablative HSCT from HLA-matched siblings with fludarabine, busulfan, and anti-thymocyte globulin as their preparative regimen. Graft-versus-host disease prophylaxis consisted of cyclosporine. RESULTS: Both children are >2 years after their HSCT. One remains a mixed chimera, and the other shows 100% donor chimerism. Both children are now free of infections and are no longer dependent on intravenous gammaglobulin. Both show response to immunizations. Both have had resolution of their cholangiopathy. CONCLUSIONS: Nonmyeloablative HSCT from HLA-matched siblings can offer immune reconstitution without hepatotoxicity in patients with X-HIM syndrome and preexisting cholangiopathy. Even with stable mixed chimerism after allogeneic HSCT, patients may be able to enjoy a normal phenotype. Nonmyeloablative HSCT warrants additional study in X-HIM syndrome. PMID- 14754982 TI - Gastroesophageal reflux: a critical review of its role in preterm infants. AB - There is widespread concern about gastroesophageal reflux (GER) in preterm infants. This article reviews the evidence for this concern. GER is common in infants, which is related to their large fluid intake (corresponding to 14 L/day in an adult) and supine body position, resulting in the gastroesophageal junction's being constantly "under water." pH monitoring, the standard for reflux detection, is of limited use in preterm infants whose gastric pH is >4 for 90% of the time. New methods such as the multiple intraluminal impedance technique and micromanometric catheters may be promising alternatives but require careful evaluation before applying them to clinical practice. A critical review of the evidence for potential sequelae of GER in preterm infants shows that 1) apnea is unrelated to GER in most infants, 2) failure to thrive practically does not occur with GER, and 3) a relationship between GER and chronic airway problems has not yet been confirmed in preterm infants. Thus, there is currently insufficient evidence to justify the apparently widespread practice of treating GER in infants with symptoms such as recurrent apnea or regurgitation or of prolonging their hospital stay, unless there is unequivocal evidence of complications, eg, recurrent aspiration or cyanosis during vomiting. Objective criteria that help to identify those presumably few infants who do require treatment for GER disease are urgently needed. PMID- 14754983 TI - European Society for Paediatric Endocrinology/Lawson Wilkins Pediatric Endocrine Society consensus statement on diabetic ketoacidosis in children and adolescents. PMID- 14754984 TI - Termination of life support after severe child abuse: the role of a guardian ad litem. AB - Discontinuation of life-sustaining interventions often raises ethical concerns. In cases of severe child abuse with poor prognosis for recovery, accused parents may have a conflict of interest regarding medical decision-making for their child, because the outcome of such decisions may impact legal charges filed against them. The recently issued American Academy of Pediatrics guidelines for addressing such cases recommended the appointment of a guardian ad litem for medical decision-making. We present the case of an 8-month-old infant who was abused severely by her father, resulting in a persistent vegetative state. We describe our experience with appointing a guardian ad litem and the ethical issues involved. PMID- 14754985 TI - Adolescent twin sisters with severe acute respiratory syndrome (SARS). AB - A novel coronavirus-associated communicable respiratory disease, severe acute respiratory syndrome (SARS), spread worldwide after an outbreak in Guangdong Province of the People's Republic of China in November 2002. Since late February 2003, there has been an epidemic in Hong Kong involving both adult and pediatric patients. The clinical course, intensive care, and outcome of adolescent twin sisters with SARS are described. Adolescents infected with SARS may develop severe illness as adults, and close monitoring for disease progression in terms of both clinical and radiologic deterioration is warranted. PMID- 14754986 TI - Ultrafast magnetic resonance imaging of the neonate in a magnetic resonance compatible incubator with a built-in coil. AB - BACKGROUND: Magnetic resonance (MR) imaging of the neonate is important clinically, because this group of patients often has complex and multiple problems due to prematurity and developmental abnormalities. MR imaging usually involves moving neonates away from their controlled environment to the scanner. OBJECTIVE: In this study we present the results of our initial experience with an MR-compatible incubator used on a 1.5-T system. METHODS: Seven neonates were imaged at 1.5 T without sedation or anesthesia. Images were obtained by using single-shot fast spin echo, 3-dimensional Fourier transfer gradient echo, and diffusion-weighted sequences. In 4 cases, time-of-flight angiography was performed. RESULTS: All 7 neonates were stable throughout the scan time (10-21 minutes). Experienced observers graded the images for quality, and all were graded excellent or good. In no case was the image quality poor. CONCLUSION: Neonates can be imaged safely by using an MR-compatible incubator and fast image sequences. This method should allow neonates to be imaged by MR in sites at which a dedicated neonatal MR scanner is not available. PMID- 14754987 TI - Recognition of DNA substrates by T4 bacteriophage polynucleotide kinase. AB - T4 phage polynucleotide kinase (PNK) displays 5'-hydroxyl kinase, 3'-phosphatase and 2',3'-cyclic phosphodiesterase activities. The enzyme phosphorylates the 5' hydroxyl termini of a wide variety of nucleic acid substrates, a behavior studied here through the determination of a series of crystal structures with single stranded (ss)DNA oligonucleotide substrates of various lengths and sequences. In these structures, the 5' ribose hydroxyl is buried in the kinase active site in proper alignment for phosphoryl transfer. Depending on the ssDNA length, the first two or three nucleotide bases are well ordered. Numerous contacts are made both to the phosphoribosyl backbone and to the ordered bases. The position, side chain contacts and internucleotide stacking interactions of the ordered bases are strikingly different for a 5'-GT DNA end than for a 5'-TG end. The base preferences displayed at those positions by PNK are attributable to differences in the enzyme binding interactions and in the DNA conformation for each unique substrate molecule. PMID- 14754988 TI - Targeting Alzheimer's disease genes with RNA interference: an efficient strategy for silencing mutant alleles. AB - Tau and amyloid precursor protein (APP) are key proteins in the pathogenesis of sporadic and inherited Alzheimer's disease. Thus, developing ways to inhibit production of these proteins is of great research and therapeutic interest. The selective silencing of mutant alleles, moreover, represents an attractive strategy for treating inherited dementias and other dominantly inherited disorders. Here, using tau and APP as model targets, we describe an efficient method for producing small interfering RNA (siRNA) against essentially any targeted region of a gene. We then use this approach to develop siRNAs that display optimal allele-specific silencing against a well-characterized tau mutation (V337M) and the most widely studied APP mutation (APPsw). The allele specific RNA duplexes identified by this method then served as templates for constructing short hairpin RNA (shRNA) plasmids that successfully silenced mutant tau or APP alleles. These plasmids should prove useful in experimental and therapeutic studies of Alzheimer's disease. Our results suggest guiding principles for the production of allele-specific siRNA, and the general method described here should facilitate the production of gene-specific siRNAs. PMID- 14754989 TI - Proteomic characterization of the interstitial fluid perfusing the breast tumor microenvironment: a novel resource for biomarker and therapeutic target discovery. AB - Clinical cancer proteomics aims at the identification of markers for early detection and predictive purposes, as well as to provide novel targets for drug discovery and therapeutic intervention. Proteomics-based analysis of traditional sources of biomarkers, such as serum, plasma, or tissue lyzates, has resulted in a wealth of information and the finding of several potential tumor biomarkers. However, many of these markers have shown limited usefulness in a clinical setting, underscoring the need for new clinically relevant sources. Here we present a novel and highly promising source of biomarkers, the tumor interstitial fluid (TIF) that perfuses the breast tumor microenvironment. We collected TIFs from small pieces of freshly dissected invasive breast carcinomas and analyzed them by two-dimensional polyacrylamide gel electrophoresis in combination with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, Western immunoblotting, as well as by cytokine-specific antibody arrays. This approach provided for the first time a snapshot of the protein components of the TIF, which we show consists of more than one thousand proteins--either secreted, shed by membrane vesicles, or externalized due to cell death--produced by the complex network of cell types that make up the tumor microenvironment. So far, we have identified 267 primary translation products including, but not limited to, proteins involved in cell proliferation, invasion, angiogenesis, metastasis, inflammation, protein synthesis, energy metabolism, oxidative stress, the actin cytoskeleton assembly, protein folding, and transport. As expected, the TIF contained several classical serum proteins. Considering that the protein composition of the TIF reflects the physiological and pathological state of the tissue, it should provide a new and potentially rich resource for diagnostic biomarker discovery and for identifying more selective targets for therapeutic intervention. PMID- 14754991 TI - Intense exercise up-regulates Na+,K+-ATPase isoform mRNA, but not protein expression in human skeletal muscle. AB - Characterization of expression of, and consequently also the acute exercise effects on, Na(+),K(+)-ATPase isoforms in human skeletal muscle remains incomplete and was therefore investigated. Fifteen healthy subjects (eight males, seven females) performed fatiguing, knee extensor exercise at approximately 40% of their maximal work output per contraction. A vastus lateralis muscle biopsy was taken at rest, fatigue and 3 and 24 h postexercise, and analysed for Na(+),K(+)-ATPase alpha(1), alpha(2), alpha(3), beta(1), beta(2) and beta(3) mRNA and crude homogenate protein expression, using Real-Time RT-PCR and immunoblotting, respectively. Each individual expressed gene transcripts and protein bands for each Na(+),K(+)-ATPase isoform. Each isoform was also expressed in a primary human skeletal muscle cell culture. Intense exercise (352 +/- 69 s; mean +/-s.e.m.) immediately increased alpha(3) and beta(2) mRNA by 2.4- and 1.7 fold, respectively (P < 0.05), whilst alpha(1) and alpha(2) mRNA were increased by 2.5- and 3.5-fold at 24 h and 3 h postexercise, respectively (P < 0.05). No significant change occurred for beta(1) and beta(3) mRNA, reflecting variable time-dependent responses. When the average postexercise value was contrasted to rest, mRNA increased for alpha(1), alpha(2), alpha(3), beta(1), beta(2) and beta(3) isoforms, by 1.4-, 2.2-, 1.4-, 1.1-, 1.0- and 1.0-fold, respectively (P < 0.05). However, exercise did not alter the protein abundance of the alpha(1) alpha(3) and beta(1)-beta(3) isoforms. Thus, human skeletal muscle expresses each of the Na(+),K(+)-ATPase alpha(1), alpha(2), alpha(3), beta(1), beta(2) and beta(3) isoforms, evidenced at both transcription and protein levels. Whilst brief exercise increased Na(+),K(+)-ATPase isoform mRNA expression, there was no effect on isoform protein expression, suggesting that the exercise challenge was insufficient for muscle Na(+),K(+)-ATPase up-regulation. PMID- 14754993 TI - Segmentation induced by intraluminal fatty acid in isolated guinea-pig duodenum and jejunum. AB - Small intestinal movements depend on the composition of the chyme with mixing predominating at high nutrient levels and propulsion being prevalent at low nutrient levels. The mechanisms coupling nutrients to motility are unknown. We used computer analysis of video recordings of isolated guinea-pig duodenum, jejunum and ileum to examine movements induced by a fatty acid, decanoic acid. Increasing intraluminal pressure past a threshold using control saline consistently evoked propulsive reflexes: lumen-occluding constrictions appeared at the oral end propagating at 20.4 +/- 2.4 mm s(-1) (mean +/-s.d., jejunum) to the anal end before being repeated until the intraluminal pressure was returned to control. Subthreshold pressure increases sometimes evoked a transient series of constrictions appearing at the oral end and propagating anally at 18.4 +/- 4.7 mm s(-1) (jejunum). At basal pressures, decanoic acid dose-dependently induced motor activity consisting of 40-60 s episodes of constrictions separated by 40 200 s periods of quiescence and lasting up to 2 h. Five contraction patterns were identified within episodes including localized stationary constrictions; constrictions that propagated slowly (5-8 mm s(-1)) for short distances orally or anally; and constrictions that propagated orally or anally for the length of the preparation at 14-20 mm s(-1). Decanoic acid induced motor activity was reversibly abolished by tetrodotoxin (3 microm), hyoscine (1 microm) and hexamethonium (100 microm), but was insensitive to blockade of P2 purinoceptors by PPADS (60 microm). Thus, decanoic acid induces motor activity equivalent to segmentation in guinea-pig small intestine in vitro and this depends on intrinsic neural pathways. PMID- 14754992 TI - Effect of acute and chronic ascorbic acid on flow-mediated dilatation with sedentary and physically active human ageing. AB - Peripheral conduit artery flow-mediated dilatation decreases with ageing in humans. The underlying mechanisms and efficacy of preventive strategies are unknown. Brachial artery flow-mediated dilatation was determined at baseline and after ascorbic acid (vitamin C) intravenous infusion and chronic supplementation (500 mg day(-1) for 30 days) in three groups of healthy men: young sedentary (n= 11; 25 +/- 1 years, mean +/-s.e.m.), older sedentary (n= 9; 64 +/- 2), and older endurance-exercise trained (n= 9; 64 +/- 2). At baseline, flow-mediated dilatation (normalized for the hyperaemic stimulus) was approximately 45% lower in the older (0.015 +/- 0.001) versus young (0.028 +/- 0.004) sedentary men (P < 0.01), but was preserved in older exercising men (0.028 +/- 0.004). Ascorbic acid infusion increased plasma concentrations > 15-fold in all groups and restored flow-mediated dilatation in the sedentary older men (to 0.023 +/- 0.002; P > 0.1 versus other groups), with no effects in the other two groups. Oral ascorbic acid supplementation did not affect flow-mediated dilatation in any group. Brachial artery endothelium-independent dilatation (sublingual nitroglycerin) did not differ among the groups at baseline nor change with ascorbic acid administration. These results provide the first evidence for an important role of oxidative stress in both the impairment in peripheral conduit artery flow-mediated dilatation with sedentary human ageing and the preservation of flow-mediated dilatation with physically active ageing. PMID- 14754994 TI - Identification of an N-terminal amino acid of the CLC-3 chloride channel critical in phosphorylation-dependent activation of a CaMKII-activated chloride current. AB - CLC-3, a member of the CLC family of chloride channels, mediates function in many cell types in the body. The multifunctional calcium-calmodulin-dependent protein kinase II (CaMKII) has been shown to activate recombinant CLC-3 stably expressed in tsA cells, a human embryonic kidney cell line derivative, and natively expressed channel protein in a human colonic tumour cell line T84. We examined the CaMKII-dependent regulation of CLC-3 in a smooth muscle cell model as well as in the human colonic tumour cell line, HT29, using whole-cell voltage clamp. In CLC-3-expressing cells, we observed the activation of a Cl(-) conductance following intracellular introduction of the isolated autonomous CaMKII into the voltage-clamped cell via the patch pipette. The CaMKII-dependent Cl(-) conductance was not observed following exposure of the cells to 1 microm autocamtide inhibitory peptide (AIP), a selective inhibitor of CaMKII. Arterial smooth muscle cells express a robust CaMKII-activated Cl(-) conductance; however, CLC-3(-/-) cells did not. The N-terminus of CLC-3, which contains a CaMKII consensus sequence, was phosphorylated by CaMKII in vitro, and mutation of the serine at position 109 (S109A) abolished the CaMKII-dependent Cl(-) conductance, indicating that this residue is important in the gating of CLC-3 at the plasma membrane. PMID- 14754996 TI - Inositol-1,4,5-trisphosphate-dependent Ca(2+) signalling in cat atrial excitation contraction coupling and arrhythmias. AB - Inositol-1,4,5-trisphosphate (IP(3))-dependent Ca(2+) release represents the major Ca(2+) mobilizing pathway responsible for diverse functions in non excitable cells. In the heart, however, its role is largely unknown or controversial. In intact cat atrial myocytes, endothelin (ET-1) increased basal [Ca(2+)](i) levels, enhanced action potential-evoked [Ca(2+)](i) transients, caused [Ca(2+)](i) transients with alternating amplitudes (Ca(2+) alternans), and facilitated spontaneous Ca(2+) release from the sarcoplasmic reticulum (SR) in the form of Ca(2+) sparks and arrhythmogenic Ca(2+) waves. These effects were prevented by the IP(3) receptor (IP(3)R) blocker aminoethoxydiphenyl borate (2 APB), suggesting the involvement of IP(3)-dependent SR Ca(2+) release. In saponin permeabilized myocytes IP(3) and the more potent IP(3)R agonist adenophostin increased basal [Ca(2+)](i) and the frequency of spontaneous Ca(2+) sparks. In the presence of tetracaine to eliminate Ca(2+) release from ryanodine receptor (RyR) SR Ca(2+) release channels, IP(3) and adenophostin triggered unique elementary, non-propagating IP(3)R-dependent Ca(2+) release events with amplitudes and kinetics that were distinctly different from classical RyR dependent Ca(2+) sparks. The effects of IP(3) and adenophostin were prevented by heparin and 2-APB. The data suggest that IP(3)-dependent Ca(2+) release increases [Ca(2+)](i) in the vicinity of RyRs and thus facilitates Ca(2+)-induced Ca(2+) release during excitation-contraction coupling. It is concluded that in the adult mammalian atrium IP(3)-dependent Ca(2+) release enhances atrial Ca(2+) signalling and exerts a positive inotropic effect. In addition, by facilitating Ca(2+) release, IP(3) may also be an important component in the development of Ca(2+) mediated atrial arrhythmias. PMID- 14754995 TI - Induction, modification and accumulation of HSP70s in the rat liver after acute exercise: early and late responses. AB - Liver cells synthesize HSP72, the cytosolic highly stress-inducible member of the 70 kDa family of heat-shock proteins (HSP70s), in response to acute exercise. This study was aimed at obtaining further insight into the physiological relevance of the hepatic stress response to exercise by investigating the induction and long-term maintenance of increased levels of HSP70s of the HSP and glucose-regulated protein (GRP) families, their post-translational modifications during or after exercise and the possible relation of HSP induction to oxidative stress. In a running rat model, acute exercise activated the synthesis and accumulation of HSP72, GRP75 and GRP78 in liver cells, pointing towards a multifactorial origin of this response. A peak HSP72 accumulation was observed shortly after exercise as a result of transcriptional activation. HSP72 was reduced shortly after exercise preceding the disappearance of its mRNA. Two further waves of HSP72 accumulation peaked 8 and 48 h after exercise without transcriptional activation. A transient increase in the proportion of acidic variants of HSP72 and HSP73 was also observed shortly after exercise as a result, at least in part, of protein phosphorylation. Free and protein-bound lipid peroxidation derivatives (TBARS) showed a tendency to increase in the early post exercise and the free-to-protein-bound TBARS ratio decreased significantly after 2 h. During the early post-exercise period, protein-bound TBARS correlated positively with HSP72 and 73, but not with GRP75 or GRP78. Altogether, the reported results indicate that the early induction and post-translational modification of HSP70s in liver cells following exercise is a preliminary step of a series of long-lasting HSP70-related events, possibly designed to preserve liver cell homeostasis and to help provide a concerted response of the whole organism to physical stress. PMID- 14754997 TI - Interstitial cells of Cajal are functionally innervated by excitatory motor neurones in the murine intestine. AB - Recent studies have demonstrated that intramuscular interstitial cells of Cajal (ICC) are preferential targets for neurotransmission in the stomach. Terminals of enteric motor neurones also form tight, synaptic-like contacts with ICC in the small intestine and colon, but little is known about the role of these cells in neurotransmission. ICC at the deep muscular plexus (ICC-DMP) of the small intestine express neurokinin 1 receptors (NK1R) and internalize these receptors in response to exogenous substance P. We used NK1R internalization as an assay of functional innervation of ICC-DMP in the murine small intestine. Under basal conditions NK1R-like immunoreactivity (NK1R-LI) was mainly observed in ICC-DMP (519 cells counted, 100% were positive) and myenteric neurones. ICC-DMP were closely apposed to substance P-containing nerve fibres. Of 338 ICC-DMP examined, 65% were closely associated with at least one substance P-positive nerve fibre, 32% were associated with at least two, 2% were associated with more than two nerve fibres and 1% with none. After electrical field stimulation (EFS, 10 Hz; 1 min) NK1R-LI was internalized in more than 80% of ICC-DMP, as compared to 10% of cells before EFS. Internalization of NK1R was not observed in myenteric ICC or smooth muscle cells in response to nerve stimulation. Internalization of NK1R-LI was blocked by the specific NK1 receptor antagonist WIN 62577 (1 microm) and by tetrodotoxin (0.3 microm), suggesting that internalization resulted from stimulation of receptors with neurally released neurokinins. These data suggest that ICC-DMP are primary targets for neurokinins released from enteric motor neurones in the intestine. PMID- 14754998 TI - Kinetics of Mg2+ unblock of NMDA receptors: implications for spike-timing dependent synaptic plasticity. AB - The time course of Mg(2+) block and unblock of NMDA receptors (NMDARs) determines the extent they are activated by depolarization. Here, we directly measure the rate of NMDAR channel opening in response to depolarizations at different times after brief (1 ms) and sustained (4.6 s) applications of glutamate to nucleated patches from neocortical pyramidal neurons. The kinetics of Mg(2+) unblock were found to be non-instantaneous and complex, consisting of a prominent fast component (time constant approximately 100 micros) and slower components (time constants 4 and approximately 300 ms), the relative amplitudes of which depended on the timing of the depolarizing pulse. Fitting a kinetic model to these data indicated that Mg(2+) not only blocks the NMDAR channel, but reduces both the open probability and affinity for glutamate, while enhancing desensitization. These effects slow the rate of NMDAR channel opening in response to depolarization in a time-dependent manner such that the slower components of Mg(2+) unblock are enhanced during depolarizations at later times after glutamate application. One physiological consequence of this is that brief depolarizations occurring earlier in time after glutamate application are better able to open NMDAR channels. This finding has important implications for spike-timing dependent synaptic plasticity (STDP), where the precise (millisecond) timing of action potentials relative to synaptic inputs determines the magnitude and sign of changes in synaptic strength. Indeed, we find that STDP timing curves of NMDAR channel activation elicited by realistic dendritic action potential waveforms are narrower than expected assuming instantaneous Mg(2+) unblock, indicating that slow Mg(2+) unblock of NMDAR channels makes the STDP timing window more precise. PMID- 14754999 TI - Propagation of pacemaker activity in the guinea-pig antrum. AB - Cyclical periods of depolarization (slow waves) underlie peristaltic contractions involved in mixing and emptying of contents in the gastric antrum. Slow waves originate from a myenteric network of interstitial cells of Cajal (ICC-MY). In this study we have visualized the sequence and propagation of Ca(2+) transients associated with pacemaker potentials in the ICC network and longitudinal (LM) and circular muscle (CM) layers of the isolated guinea-pig gastric antrum. Gastric antrum was dissected to reveal the ICC-MY network, loaded with Fluo-4 AM and activity was monitored at 37 degrees C. Ca(2+) waves propagated throughout the ICC-MY network at an average velocity of 3.24 +/- 0.12 mm s(-1) at a frequency of 4.87 +/- 0.16 cycles min(-1) (n= 4). The propagation of the Ca(2+) wave often appeared 'step-like', with separate regions of the network being activated after variable delays. The direction of propagation was highly variable (Delta angle of propagation 44.3 +/- 10.9 deg per cycle) and was not confined to the axes of the longitudinal or circular muscle. Ca(2+) waves appeared to spread out radially from the site of initiation. The initiating Ca(2+) wave in ICC-MY was correlated to secondary Ca(2+) waves in intramuscular interstitial cells of Cajal, ICC-IM, and smooth muscle cells, and the local distortion (contraction) in a field of view. TTX (1 microm) had little effect on slow wave or pacemaker potential activity, but 2-APB (50 microm) blocked all Ca(2+) waves, indicating a pivotal role for intracellular Ca(2+) stores. Nicardipine (2 microm) eliminated the Ca(2+) transient generated by smooth muscle, but did not affect the fast upstroke associated with ICC-MY. These results indicate that slow waves follow a sequence of activation, beginning with the ICC-MY and ICC-IM network, followed later by a sustained Ca(2+) transient in the muscle layers that is responsible for contraction. PMID- 14755001 TI - Aerobic exercise before diving reduces venous gas bubble formation in humans. AB - We have previously shown in a rat model that a single bout of high-intensity aerobic exercise 20 h before a simulated dive reduces bubble formation and after the dive protects from lethal decompression sickness. The present study investigated the importance of these findings in man. Twelve healthy male divers were compressed in a hyperbaric chamber to 280 kPa at a rate of 100 kPa min(-1) breathing air and remaining at pressure for 80 min. The ascent rate was 9 m min( 1) with a 7 min stop at 130 kPa. Each diver underwent two randomly assigned simulated dives, with or without preceding exercise. A single interval exercise performed 24h before the dive consisted of treadmill running at 90% of maximum heart rate for 3 min, followed by exercise at 50% of maximum heart rate for 2 min; this was repeated eight times for a total exercise period of 40 min. Venous gas bubbles were monitored with an ultrasonic scanner every 20 min for 80 min after reaching surface pressure. The study demonstrated that a single bout of strenuous exercise 24h before a dive to 18 m of seawater significantly reduced the average number of bubbles in the pulmonary artery from 0.98 to 0.22 bubbles cm(-2)(P= 0.006) compared to dives without preceding exercise. The maximum bubble grade was decreased from 3 to 1.5 (P= 0.002) by pre-dive exercise, thereby increasing safety. This is the first report to indicate that pre-dive exercise may form the basis for a new way of preventing serious decompression sickness. PMID- 14755000 TI - Spatiotemporal patterning of IP3-mediated Ca2+ signals in Xenopus oocytes by Ca2+ binding proteins. AB - Ca(2+)-binding proteins (CaBPs) are expressed in a highly specific manner across many different cell types, yet the physiological basis underlying their selective distribution patterns remains unclear. We used confocal line-scan microscopy together with photo-release of IP(3) in Xenopus oocytes to investigate the actions of mobile cytosolic CaBPs on the spatiotemporal properties of IP(3) evoked Ca(2+) signals. Parvalbumin (PV), a CaBP with slow Ca(2+)-binding kinetics, shortened the duration of IP(3)-evoked Ca(2+) signals and 'balkanized' global responses into discrete localized events (puffs). In contrast, calretinin (CR), a presumed fast buffer, prolonged Ca(2+) responses and promoted 'globalization' of spatially uniform Ca(2+) signals at high [IP(3)]. Oocytes loaded with CR or PV showed Ca(2+) puffs following photolysis flashes that were subthreshold in controls, and the spatiotemporal properties of these localized events were differentially modulated by PV and CR. In comparison to results we previously obtained with exogenous Ca(2+) buffers, PV closely mimicked the actions of the slow buffer EGTA, whereas CR showed important differences from the fast buffer BAPTA. Most notably, puffs were never observed after loading BAPTA, and this exogenous buffer did not show the marked sensitization of IP(3) action evident with CR. The ability of Ca(2+) buffers and CaBPs with differing kinetics to fine-tune both global and local intracellular Ca(2+) signals is likely to have significant physiological implications. PMID- 14755002 TI - Central CRF, urocortins and stress increase colonic transit via CRF1 receptors while activation of CRF2 receptors delays gastric transit in mice. AB - Recently characterized selective agonists and developed antagonists for the corticotropin releasing factor (CRF) receptors are new tools to investigate stress-related functional changes. The influence of mammalian CRF and related peptides injected intracerebroventricularly (i.c.v.) on gastric and colonic motility, and the CRF receptor subtypes involved and their role in colonic response to stress were studied in conscious mice. The CRF(1)/CRF(2) agonists rat urocortin 1 (rUcn 1) and rat/human CRF (r/h CRF), the preferential CRF(1) agonist ovine CRF (oCRF), and the CRF(2) agonist mouse (m) Ucn 2, injected i.c.v. inhibited gastric emptying and stimulated distal colonic motor function (bead transit and defecation) while oCRF(9-33)OH (devoid of CRF receptor affinity) showed neither effects. mUcn 2 injected peripherally had no colonic effect. The selective CRF(2) antagonist astressin(2)-B (i.c.v.), at a 20 : 1 antagonist: agonist ratio, blocked i.c.v. r/hCRF and rUcn 1 induced inhibition of gastric transit and reduced that of mUcn 2, while the CRF(1) antagonist NBI-35965 had no effect. By contrast, the colonic motor stimulation induced by i.c.v. r/hCRF and rUcn 1 and 1h restraint stress were antagonized only by NBI-35965 while stimulation induced by mUcn 2 was equally blocked by both antagonists. None of the CRF antagonists injected i.c.v. alone influenced gut transit. These data establish in mice that brain CRF(1) receptors mediate the stimulation of colonic transit induced by central CRF, urocortins (1 and 2) and restraint stress, while CRF(2) receptors mediate the inhibitory actions of these peptides on gastric transit. PMID- 14755003 TI - The sense of movement elicited by transcranial magnetic stimulation in humans is due to sensory feedback. AB - It has been claimed that transcranial magnetic stimulation (TMS) of the human motor cortex can produce a sense of movement of the contralateral hand, even when the hand is paralysed. The sense of movement was equated with a 'corollary discharge', a nulling mechanism originally posited for maintaining constancy of the visual field during eye movements. Our experiments were designed to test whether the sensation that accompanies TMS-evoked finger movements is generated centrally or whether it arises as a result of sensory feedback. Matched twitches of the left and right fingers were elicited either by bilateral electrical stimulation of forearm extensor muscles, or by a combination of TMS of left motor cortex (eliciting twitches of the right forefinger), and electrical stimulation of the left forearm muscles (eliciting twitches of the left forefinger). The time interval between stimuli activating left and right twitches was varied randomly (range +/- 90 ms) from trial to trial. Subjects reported whether they sensed that the left or the right movement occurred first, or if they could detect no difference. The left and right movements evoked by bilateral electrical stimulation of muscles were sensed as near simultaneous when there was zero delay between them. When TMS was applied in conjunction with unilateral muscle stimulation, the TMS-evoked movement was felt, on average, 20 ms after the movement evoked by muscle stimulation. Similar results were obtained when the skin under the cathodal electrodes was anaesthetized. Since the TMS-evoked movements were felt later rather than earlier than the electrically evoked movements, the results do not support the idea that the sensation of movement was elicited centrally by TMS. Rather, they favour sensory feedback as the source of the sense of movement. The earlier perception of electrically evoked versus TMS evoked movements was probably due to earlier sensory responses in the periphery rather than a suppression of the excitability of somatosensory cortex. PMID- 14755004 TI - Differential conditioned place preference responses to endomorphin-1 and endomorphin-2 microinjected into the posterior nucleus accumbens shell and ventral tegmental area in the rat. AB - An unbiased conditioned place preference (CPP) paradigm was used to evaluate the reward effects of endogenous mu-opioid receptor ligands endomorphin-1 (EM-1) and endomorphin-2 (EM-2) from the mesolimbic posterior nucleus accumbens (Acb) shell and the ventral tegmental area (VTA) in CD rats. EM-1 (1.6-8.1 nmol) microinjected into posterior Acb shell produced CPP, whereas EM-2 (8.7-17.5 nmol) given into the same Acb shell produced conditioned place aversion (CPA). EM-1 (1.6-16.3 nmol) microinjected into the VTA produced CPP, whereas EM-2 (8.7 and 17.5 nmol) given into the same VTA site did not produce any effect, but at a high dose (35 nmol) produced CPP. EM-1 (3.3 nmol) or EM-2 (17.5 nmol) microinjected into the nigrostriatal substantia nigra was not significantly different from vehicle-injected groups. D-Phe-Cys-Tyr-D-Trp-Orn-Thr-Pen-Thr-NH(2) (CTOP) at 94.13 pmol or 3-methoxynaltrexone at 0.64 pmol microinjected into the posterior Acb shell blocked EM-1-induced CPP and EM-2-induced CPA. At a higher dose, CTOP (941.3 pmol) and 3-methoxynaltrexone (6.4 pmol) produced CPA and CPP, respectively. Coadministration with antiserum against dynorphin A(1-17) (Dyn) (10 microg) microinjected into the posterior Acb shell blocked EM-2-induced CPA. However, it did not affect EM-1-induced CPP. It is concluded that EM-1 and EM-2 produce site-dependent CPP and CPA, respectively, by stimulation of different subtypes of mu-opioid-receptors; stimulation of one subtype of mu-opioid-receptor at the posterior Acb shell and VTA by EM-1 induces CPP, whereas stimulation of another subtype of mu-opioid receptor at the posterior Acb shell, but not the VTA, by EM-2 induces the release of Dyn to produce CPA. PMID- 14755005 TI - Dopaminergic agonists and muscarinic antagonists improve lateralization in hemiparkinsonian rats in a novel exploratory Y-maze. AB - Parkinson's disease (PD) is characterized by the degeneration of nigrostriatal dopaminergic neurons. Its primary clinical symptoms are akinesia, tremor, and rigidity, which usually start from one side, resembling the lateralization in hemiparkinsonian rats having 6-hydroxydopamine-induced unilateral lesion of the medial forebrain bundle. A novel exploratory Y-maze was designed to detect the lateralization of hemiparkinsonian rats in terms of biased turns in the maze. Dopamine agonists levodopa (L-3,4-dihydroxyphenylalanine, 10-30 mg/kg) and apomorphine (0.1-0.3 mg/kg), but not methamphetamine (0.5-2 mg/kg), improved the lateralization in the rat model. However, high doses of the dopamine agonists, 30 and 0.3 mg/kg, respectively, caused small movements in the arms that seemed to parallel the increase in counts per turn in the Y-maze. Interestingly, the muscarinic antagonists trihexyphenidyl and scopolamine improved lateralization moderately, while increasing total turns, an index of locomotive activity. (-)-5 Methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imine maleate (MK-801) (0.3 mg/kg), an N-methyl-D-aspartate (NMDA) glutamate receptor antagonist, increased total counts, but did not alleviate the lateralization. The alpha2-adrenoceptor antagonist idazoxan (1 and 10 mg/kg) and 2,3-dihydroxy-6-nitro-7 sulfamoylbenzo(f)quinoxaline (1 and 3 mg/kg), a non-NMDA glutamate receptor antagonist, did not affect any of the indices. These findings suggest that the clinical action of drugs on unbalanced movement in PD could be predicted by measuring their effects on lateralization of the 6-hydroxydopamine-lesioned rat model in this exploratory Y-maze. PMID- 14755006 TI - Effects of N-substituted analogs of benztropine: diminished cocaine-like effects in dopamine transporter ligands. AB - Previous studies demonstrated that analogs of benztropine (BZT) possess high affinity for the dopamine transporter, inhibit dopamine uptake, but generally have behavioral effects different from those of cocaine. One hypothesis is that muscarinic-M(1) receptor actions interfere with cocaine-like effects. Several tropane-nitrogen substitutions of 4',4"-diF-BZT have reduced M(1) affinity compared with the CH(3)-analog (AHN 1-055; 3alpha-[bis-(4 fluorophenyl)methoxy]tropane). All of the compounds displaced [(3)H]WIN 35,428 (2beta-carbomethoxy-3beta-(4-fluorophenyl)tropane) binding with affinities ranging from 11 to 108 nM. Affinities at norepinephrine ([(3)H]nisoxetine) and serotonin ([(3)H]citalopram) transporters ranged from 457 to 4810 and 376 to 3260 nM, respectively, and at muscarinic M(1) receptors ([(3)H]pirenzepine) from 11.6 (AHN 1-055) to higher values, reaching 1030 nM for the other BZT-analogs. Cocaine and AHN 1-055 produced dose-related increases in locomotor activity in mice, with AHN 1-055 less effective than cocaine. The other compounds were ineffective in stimulating activity. In rats discriminating cocaine (29 micromol/kg i.p.) from saline, WIN 35,428 fully substituted for cocaine, whereas AHN 1-055 produced a maximal substitution of 79%. None of the other analogs fully substituted for cocaine. WIN 35,428 produced dose-related leftward shifts in the cocaine dose effect curve, whereas selected BZT analogs produced minimal changes in the effects of cocaine. The results suggest that reducing M(1) affinity of 4',4"-diF BZT with N-substitutions reduces effectiveness in potentiating the effects of cocaine. Furthermore, although the BZT-analogs bind with high affinity at the dopamine transporter, their behavioral effects differ from those of cocaine. These compounds have reduced efficacy compared with cocaine, a long duration of action, and may serve as leads for the development of medications to treat cocaine abuse. PMID- 14755007 TI - Gastric tolerability and prolonged prostaglandin inhibition in the brain with a nitric oxide-releasing flurbiprofen derivative, NCX-2216 [3-[4-(2-fluoro-alpha methyl-[1,1'-biphenyl]-4-acetyloxy)-3-methoxyphenyl]-2-propenoic acid 4-nitrooxy butyl ester]. AB - NCX-2216 [3-[4-(2-fluoro-alpha-methyl-[1,1'-biphenyl]-4-acetyloxy)-3 methoxyphenyl]-2-propenoic acid 4-nitrooxy butyl ester] is an NO-releasing flurbiprofen derivative that also contains a ferulic acid (antioxidant) moiety. NCX-2216 has been shown to be effective in reducing beta-amyloid deposition in a transgenic mouse model of Alzheimer's disease. The tolerability of this compound in the stomach and its ability to suppress prostaglandin synthesis in the brain are not known. The purpose of this study was to assess the contribution of nitric oxide (NO) and ferulic acid to the pharmacological properties of NCX-2216 versus flurbiprofen; thus, we compared their gastric tolerability and suppression of prostaglandin synthesis, peripherally and centrally. Oral flurbiprofen produced extensive gastric damage and suppressed gastric prostaglandin synthesis. In contrast, while suppressing prostaglandin production, equimolar doses of NCX-2216 did not cause detectable gastric injury. The NO-releasing moiety of NCX-2216 (but not the ferulic acid moiety) was crucial for the gastric safety of this compound. NCX-2216 substantially inhibited prostanoid synthesis despite not being detectable in plasma and despite producing only low amounts of flurbiprofen in plasma and in the brain. Inhibition of brain prostaglandin synthesis by NCX-2216 (22 mg/kg) persisted for a much longer period of time (up to 48 h) than was seen with flurbiprofen ( or =65 years of age, but older patients may have been more susceptible to dehydration, diarrhea, hypokalemia, and fatigue. Oxaliplatin in combination with infusional 5-FU/LV was approved for the treatment of patients with metastatic carcinoma of the colon or rectum whose disease has recurred or progressed during or within 6 months of completion of first-line therapy with the combination of bolus 5-FU/LV and irinotecan. Approval was based on response rate and on an interim analysis of TTP. No results are available, at this time, that demonstrate a clinical benefit, such as improvement in disease-related symptoms or survival. PMID- 14755011 TI - Recognition and management of hereditary breast cancer syndromes. AB - Clinicians should recognize the genetic syndromes that predispose to the development of breast cancer so that patients may be afforded the opportunity to have genetic testing to assist them and their family members in making medical management decisions. Approximately 80%-90% of hereditary breast cancer cases are caused by mutations in the BRCA1 and BRCA2 genes. Other important clinical genetic predispositions include Cowden syndrome, Li-Fraumeni syndrome, Peutz Jeghers syndrome, and ataxia-telangiectasia. The key to identifying women who are at risk for a hereditary breast cancer lies in obtaining an adequate, three generation family history, including ethnic background. For unaffected women, breast cancer risks can be estimated using the quantitative models of Gail and Claus, but there are limitations to these models. Other quantitative models predict the likelihood that a patient is carrying a mutated gene. Genetic testing is available at selected laboratories for each of the hereditary syndromes described, and there are three possible outcomes to testing. These outcomes and their management implications are described in detail. Clinical management options for women at high risk for breast cancer include surveillance, chemoprevention, and prophylactic surgery. Application of these principles can reduce morbidity in women with genetic predispositions to breast cancer. PMID- 14755012 TI - Anemia in stage II and III breast cancer patients treated with adjuvant doxorubicin and cyclophosphamide chemotherapy. AB - PURPOSE: The incidence and severity of prechemotherapy anemia and chemotherapy induced anemia experienced by women treated with adjuvant doxorubicin and cyclophosphamide (AC) therapy for stage II and III breast cancer are described. PATIENTS AND METHODS: Medical charts of 310 breast cancer patients who received chemotherapy at eight oncology practices during 1997 through 2001 were reviewed in this historical case series study. Prechemotherapy anemia was defined as a baseline hemoglobin value <12 g/dl. An anemic event during chemotherapy (used to define chemotherapy-induced anemia) was defined as either a drop in hemoglobin level below the threshold (< or = 10 g/dl), the receipt of a blood transfusion(s), or treatment with epoetin alfa. RESULTS: Overall, 40.0% of patients experienced moderate to severe anemia (i.e., their hemoglobin levels dropped to <10 g/dl) and 31.3% (97/310) were anemic prechemotherapy. Of the patients with mild anemia prechemotherapy, 61.9% developed moderate to severe anemia during chemotherapy. Only 47.4% (46/97) of those patients received epoetin alfa therapy during chemotherapy. Of the patients with normal prechemotherapy hemoglobin levels (> or = 12 g/dl), 88.3% developed some degree of anemia (<12 g/dl) during chemotherapy and 27.7% (59/213) developed moderate to severe anemia (<10 g/dl). Anemic events were experienced by 41.8% (89/213) of the patients with normal prechemotherapy hemoglobin levels. CONCLUSIONS: We conclude that a significant proportion (31.3%) of stage II and III breast cancer patients are anemic prechemotherapy and that the incidence of anemia increases substantially from prechemotherapy through the postchemotherapy period. This evidence appears to warrant earlier evaluation of anemia and an intervention in the prechemotherapy stage. PMID- 14755013 TI - Topotecan dosing guidelines in ovarian cancer: reduction and management of hematologic toxicity. AB - Topotecan dosing considerations and alternative dosing schedules to reduce and manage myelosuppression during the treatment of relapsed ovarian cancer were reviewed. The myelosuppression patterns from phase I, II, and III clinical trials were analyzed to evaluate the degree of hematologic toxicity and to determine risk factors predictive of myelosuppression. Additionally, recent publications of alternative topotecan doses and schedules were examined. Extent of prior therapy, prior platinum therapy (particularly carboplatin), advanced age, impaired renal function, and prior radiation therapy were identified as potential risk factors for greater hematologic toxicity after topotecan therapy. Reducing the starting topotecan dose to 1.0 or 1.25 mg/m2/day is recommended to reduce the incidence of severe myelosuppression in high-risk individuals receiving topotecan for 5 consecutive days. Hematopoietic growth factors, transfusion therapy, and schedule adjustments may also help manage myelosuppression. Alternative schedules of 3-day or weekly dosing appear to have less myelotoxicity and are currently under evaluation. The clinical aspects of topotecan-related myelosuppression and results from clinical trials indicate that the dose, and possibly the dosing schedule, of topotecan can be modified to reduce hematologic toxicity and improve tolerance without compromising efficacy. Prospective individualization of topotecan dosing may prevent or minimize dose-limiting myelosuppression and allow patients to achieve the maximum topotecan benefit by improving their ability to complete therapy with fewer treatment delays. Ongoing clinical trials evaluating alternative dosing schedules with superior hematologic tolerability may facilitate the inclusion of topotecan in combination regimens for patients with ovarian cancer. Proposed topotecan dosing guidelines to reduce and manage myelosuppression are outlined. PMID- 14755014 TI - Diagnosis and treatment of cholangiocarcinoma. AB - Cholangiocarcinoma presents a formidable diagnostic and treatment challenge. The majority of patients present with unresectable disease and have a survival of less than 12 months following diagnosis. Progress has been made by the appropriate selection of patients for treatment options including resection, with the routine use of more aggressive resections in order to achieve margin-negative resections. This has resulted in longer survival times for these patients. Neoadjuvant and adjuvant therapies have, for the most part, not improved survival in patients with this tumor, and new strategies are needed to improve this line of therapy. The prognosis for unresectable patients is poor, and palliative measures should be aimed at increasing quality of life first and increasing survival second. PMID- 14755015 TI - HER1/EGFR targeting: refining the strategy. AB - Human epidermal growth factor receptor (EGFR), HER, targeting has formed the basis of extensive and growing drug development programs in various companies. However, receptor biology is often poorly explained and confusing. The HER family of four naturally occurring receptors and one tumor-specific mutant can activate signaling via a complex and sophisticated range of mechanisms, which we are only beginning to understand. HER1/EGFR downstream signaling can lead to tumor growth and development via a host of processes, including enhanced cellular proliferation, survival, and metastasis. A range of potential therapeutic targets exists within the HER signaling system, both inside and outside the cell. Monoclonal antibodies and tyrosine kinase inhibitors, acting extracellularly and intracellularly, respectively, comprise two classes of agents most advanced in clinical development or already available for use. Despite promising single-agent activity in chemotherapy-resistant patients with non-small cell lung cancer (NSCLC), disappointing results from two phase III trials of the tyrosine kinase inhibitor gefitinib in NSCLC have been of concern to some. However, many factors may have contributed to this outcome, and it is not necessarily predictive of the future usefulness of these agents. Patient characteristics, lack of patient selection, dosing schedule, and trial design may all have played roles. It is important to remember that intracellular targeting of HER is a relatively novel approach, and our knowledge of how best to optimize such treatment is still unfolding. More clinical experience is needed. PMID- 14755016 TI - The role of topotecan in the treatment of brain metastases. AB - Despite advances in the treatment of systemic malignancies, the prognosis for patients with brain metastases continues to be dismal. Because the majority of cytotoxic agents seem to be unable to penetrate the blood-brain barrier, the role of chemotherapy in the treatment of brain metastases remains controversial. However, growing amounts of both laboratory and clinical data suggest that a few of the newly developed cytotoxic agents can cross the blood-brain barrier and may have a role in the treatment of patients with brain metastases. Topotecan, a novel topoisomerase I inhibitor, freely crosses the blood-brain barrier and may be clinically effective in both the therapeutic and prophylactic settings in patients with brain metastases. Recent studies have demonstrated the antitumor activity of topotecan against brain metastases, with objective response rates ranging from 33%-63% in patients with various solid tumors. The antitumor response in the central nervous system was often greater and occurred more quickly than the systemic antitumor response to topotecan treatment. This result may be explained by the lack of exposure of brain metastases to previous cytotoxic agents, suggesting a role for topotecan in patients with brain metastases. Early studies have also suggested that topotecan, an apparent radiosensitizer, may be particularly effective in combination with radiotherapy, the current standard of care for patients with brain metastases. In addition, preliminary data suggest that topotecan in combination with temozolomide (another cytotoxic agent that can cross the blood-brain barrier) may have synergistic antitumor activity against brain metastases. This review summarizes the available preclinical and clinical evidence for the role of topotecan in the treatment of brain metastases and concludes with three case studies. PMID- 14755017 TI - Complementary and alternative therapies for cancer. AB - Many cancer patients use therapies promoted as literal alternatives to conventional medical care. Such "alternative" modalities are unproven or were studied and found worthless. These can be harmful. An even greater proportion of cancer patients uses "complementary" therapies along with mainstream cancer treatment. Most are helpful adjunctive approaches that control symptoms and enhance quality of life. This review describes alternative as well as complementary therapies commonly used today by cancer patients. Herbal remedies also are discussed. Evidence regarding the efficacy and safety of complementary/alternative medicine (CAM) is reviewed, and implications for oncologists are discussed. To encourage open communication of CAM use by patients, oncologists should be knowledgeable about the most popular remedies and know where to find reliable information for themselves and for their patients. PMID- 14755018 TI - Epoetin alfa 60,000 U once weekly followed by 120,000 U every 3 weeks increases and maintains hemoglobin levels in anemic cancer patients undergoing chemotherapy. AB - PURPOSE: Epoetin alfa administered s.c. three times weekly or once weekly increases hemoglobin (Hb) levels, decreases transfusion requirements, and improves quality of life in anemic cancer patients receiving chemotherapy. This study assessed the feasibility of using higher initial doses of once-weekly epoetin alfa followed by less frequent maintenance doses to increase and then maintain adequate Hb levels in this population. MATERIALS AND METHODS: In this open-label, nonrandomized, pilot study, anemic (baseline Hb < or = 11 g/dl) cancer patients undergoing chemotherapy received initial doses of epoetin alfa of 60,000 U s.c. once weekly to increase Hb levels by at least 2 g/dl, followed by 120,000 U s.c. every 3 weeks to maintain Hb levels. The maximum treatment duration was 24 weeks. RESULTS: The mean baseline Hb level was 10.1 +/- 0.8 g/dl (n = 20). Once-weekly dosing resulted in mean Hb level increases of 1.0 +/- 1.1 g/dl by week 4 and 2.9 +/- 1.9 g/dl by week 8; 86% and 79% of patients evaluable at week 8 and week 12, respectively, demonstrated increases of at least 2 g/dl (target Hb level of > or = 12 g/dl). Thirteen patients (65%) received at least one maintenance dose; the mean Hb level increased from 12.8 +/- 1.1 g/dl before starting maintenance therapy to 13.3 +/- 1.4 g/dl at the last maintenance week. Both dosage regimens were well tolerated. CONCLUSIONS: Once-weekly epoetin alfa at a dose of 60,000 U effectively increased Hb levels by week 8; 86% of patients achieved rises of at least 2 g/dl or Hb levels > or = 12 g/dl. Moreover, epoetin alfa at doses of 120,000 U every 3 weeks maintained or increased Hb levels. Results from this pilot study suggest that higher initial once-weekly dosing of epoetin alfa followed by less frequent maintenance dosing appears to be feasible for treating anemia in cancer patients undergoing chemotherapy. Further evaluation of these and other epoetin alfa dosage regimens is warranted. PMID- 14755019 TI - The evolving role of epoetin alfa in cancer therapy. AB - Since its initial indication as hormone-replacement therapy in the anemia of chronic kidney disease, epoetin alfa has become a mainstay of therapy for chemotherapy-related anemia. Clinical studies have shown that epoetin alfa administered once weekly or three times weekly improves hemoglobin levels, decreases transfusion requirements, and improves quality of life independent of tumor response to chemotherapy. Ongoing research is now evaluating ways to improve the response rate to epoetin alfa, the potential benefits of alternative dosing regimens and early treatment intervention, and nonanemia-related indications (e.g., cognitive impairment, asthenia). In addition, scientists are exploring the role of epoetin alfa in preventing apoptosis and ischemic brain injury, as well as its activity in other nonerythroid tissues. Thus, the role of epoetin alfa is likely to expand in the cancer setting in the coming years. PMID- 14755020 TI - The molecular perspective: DNA polymerase. PMID- 14755021 TI - Profile of Prof. Paul Calabresi. PMID- 14755022 TI - Art illuminates patients' experience at the Massachusetts General Hospital Cancer Center. PMID- 14755025 TI - HIV infection in black Caribbeans in the United Kingdom. PMID- 14755027 TI - Genital allergy. AB - Genital allergy should be considered as a possible diagnosis in all patients with genital soreness or irritation for which no infection or dermatosis can be identified and in whom symptoms remain unchanged or worsen with treatment. Type I and IV hypersensitivity reactions are most commonly encountered and can be assessed by performing skin prick testing/radioallergosorbent test (RAST) or patch testing, respectively. Type IV reactions (contact dermatitis) may sometimes prove difficult to distinguish clinically from an irritant dermatitis. This clinical review attempts to summarise key features of genital allergy for the practicing clinician. PMID- 14755028 TI - Managing recurrent bacterial vaginosis. AB - Bacterial vaginosis (BV) is the most frequently found condition of the female genital tract. It increases a woman's risk of acquiring HIV, is associated with increased complications in pregnancy, and may be involved in the pathogenesis of pelvic inflammatory disease. Yet there are many unanswered questions about its aetiology, making management of recurrent infection difficult and often idiosyncratic. This paper discusses the current knowledge and possible management of recurrent BV. PMID- 14755030 TI - Black Caribbean adults with HIV in England, Wales, and Northern Ireland: an emerging epidemic? AB - BACKGROUND: HIV is now well established in the Caribbean, with prevalence in several countries being surpassed only by those of sub-Saharan Africa. Continuing inward migration from the Caribbean and a high incidence of some bacterial STIs among Britain's black Caribbean communities, suggests a considerable potential for HIV spread. METHODS: Data from three national HIV/AIDS surveillance systems were reviewed, providing information on new HIV diagnoses, numbers accessing treatment and care services, and HIV prevalence. RESULTS: Between 1997 and 2001, 528 black Caribbean adults were newly diagnosed with HIV; 62 new diagnoses in 1997, rising to 176 in 2001. Probable heterosexual acquisition accounted for 335 (63%) infections (161 (48%) males, 174 females), and sex between men 171 (32%). Infection was acquired both in the Caribbean and in the United Kingdom. Numbers of black Caribbeans accessing treatment and care services more than doubled between 1997 (294) and 2001 (691). In 2001, 528 (76%) black Caribbeans accessing services were London residents. Among the Caribbean born previously undiagnosed heterosexuals, HIV prevalence was 0.7%; among men who have sex with men (MSM) it was 10.4%. Of those born in the Caribbean, 73% of male heterosexuals, 50% of female heterosexuals, and 65% of MSM who were previously undiagnosed left the clinic unaware of their HIV infection. CONCLUSIONS: Numbers of black Caribbean adults newly diagnosed and accessing treatment and care services in England, Wales, and Northern Ireland increased between 1997 and 2001. Despite a high prevalence of diagnosed bacterial STIs, prevalence among Caribbean born heterosexuals remains low, but it is high among MSM. Surveillance data highlight the need for targeted HIV prevention among black Caribbeans. PMID- 14755029 TI - Genital manifestations of tropical diseases. AB - Genital symptoms in tropical countries and among returned travellers can arise from a variety of bacterial, protozoal, and helminthic infections which are not usually sexually transmitted. The symptoms may mimic classic sexually transmitted infections (STIs) by producing ulceration (for example, amoebiasis, leishmaniasis), wart-like lesions (schistosomiasis), or lesions of the upper genital tract (epididymo-orchitis caused by tuberculosis, leprosy, and brucellosis; salpingitis as a result of tuberculosis, amoebiasis, and schistosomiasis). A variety of other genital symptoms less suggestive of STI are also seen in tropical countries. These include hydrocele (seen with filariasis), which can be no less stigmatising than STI, haemospermia (seen with schistosomiasis), and hypogonadism (which may occur in lepromatous leprosy). This article deals in turn with genital manifestations of filariasis, schistosomiasis, amoebiasis, leishmaniasis, tuberculosis and leprosy and gives clinical presentation, diagnosis, and treatment. PMID- 14755031 TI - The potential epidemiological impact of a genital herpes vaccine for women. AB - BACKGROUND: In two phase III vaccine trials immunisation of women previously uninfected by herpes simplex virus provided protection against genital herpes disease. In deciding policy, an evaluation of the epidemiological impact of the partial protection provided by the vaccine should be considered. METHODS: A sex and sexual activity stratified deterministic differential and partial differential equation model of the natural history of herpes simplex virus type 2 (HSV-2) and the impact of vaccination is developed and analysed. To explore the role of vaccination, the pattern of viral shedding and the transmission of infection during sexual acts within sexual partnerships are described. RESULTS: Using literature derived estimates of parameter values and assuming efficacy in only 40% of women the impact of the vaccine depends on assumptions made about its action. The vaccine has a limited impact if it only prevents disease but a more substantial impact if it reduces asymptomatic viral shedding, which it could do indirectly by preventing infection or directly by modifying the biology of the infection. Concern over the implications of a vaccine that prevents disease but has no impact on viral shedding was addressed in a worst case scenario. Here there is a modest increase in the incidence of infection in both men and women but an increase in disease prevalence in men alone, since the virus directly protects some women from disease. CONCLUSIONS: Results suggest that a herpes vaccine should be used universally and that a vaccine that only protects HSV-1-/2 women can paradoxically have a significant epidemiological impact, the scale of which depends upon changes in patterns of viral shedding. PMID- 14755032 TI - Physicians' opinions about partner notification methods: case reporting, patient referral, and provider referral. AB - BACKGROUND: The United States has relied upon partner notification strategies to help break the chain of infection and re-infection for sexually transmitted diseases (STD). Physicians are a vital link in the system of STD control, but little is known of physician opinions about partner notification strategies. METHODS: We collected opinions about partner notification from a national probability sample of physicians in specialties diagnosing STDs. Physicians responded to 17 questions about three relevant forms of STD partner notification: patient based referral, provider based referral, and case reporting. RESULTS: Exploratory factor analyses showed that responses for each form of partner notification could be grouped into four categories: perceived practice norms, infection control, patient relationships, and time/money. Multivariate analyses of the factors showed that physicians endorsed patient based referral most favourably and provider based referral least favourably. CONCLUSION: Physicians' opinions about partner notification strategies appear to reflect objective reality in some areas, but not in others. Strategies that improve the fit between physicians' opinions and effective notification are needed: some are discussed here. PMID- 14755033 TI - The sexual health of pupils in years 4 to 6 of primary schools in rural Tanzania. AB - BACKGROUND/OBJECTIVES: There is an urgent need for effective interventions to improve the sexual and reproductive health of adolescents. Reliable data on the sexual health of adolescents are needed to guide the development of such interventions. The aim was to describe the sexual health of pupils in years 4 to 6 of 121 rural primary schools in north western Tanzania, before the implementation of an innovative sexual health intervention in 58 of the schools. METHODS: A cross sectional survey of primary school pupils in rural Tanzania was carried out. The study population comprised pupils registered in years 4 to 6 of 121 primary schools in 20 rural communities in 1998. Basic demographic information was collected from all pupils seen. Those born before 1 January 1985 (aged approximately 14 years and over) were invited to participate in the survey, and asked about their knowledge and attitudes towards sexual health issues, and their sexual experience. A urine specimen was requested and tested for HIV, Chlamydia trachomatis (CT), Neisseria gonorrhoeae (NG) and, for females, pregnancy. RESULTS: 9283 pupils born before 1 January 1985 were enrolled and provided demographic information and a urine sample. Male pupils were significantly older than females (mean age 15.5 years v 14.8 years, p<0.001), but all other demographic characteristics were similar between the sexes. 14 (0.2%) of the enrolled pupils (four male and 10 female) were HIV positive, 83 (0.9%) were positive for CT, and 12 (0.1%) for NG. 32 female pupils (0.8%) were positive by pregnancy test. Sexual experience was reported by one fifth of primary school girls, and by almost half of boys. Only 45/114 (39%) girls with biological markers of sexual activity reported having had sex. CONCLUSIONS: HIV, CT, NG, and pregnancy were present though at relatively low levels among pupils in years 4 to 6 of primary school. A high proportion of pupils with a biological marker of sexual activity denied ever having had sex. Alternative ways of collecting sensitive data about the sexual behaviour of school pupils should be explored. PMID- 14755035 TI - Sexual behaviour of young people in international tourist resorts. AB - BACKGROUND/OBJECTIVES: Increasingly, young people travel abroad to experience nightlife in international resorts. Although media coverage of such resorts suggests high levels of sexual activity, little empirical data are currently available. We have measured: 3 year trends in sexual behaviour of young people visiting Ibiza, levels of sexual risk taking, and their relation to substance use. Additionally, in 2002 we identified levels of homosexual sex and sexual interactions between UK residents and individuals from other countries. METHODS: Data were collected from visitors to Ibiza between 2000 and 2002 just before they left the island. Information on sexual health was surveyed using a short anonymous questionnaire. RESULTS: Over half of individuals (56.0%) visiting Ibiza had sex with at least one person, with 26.2% of males and 14.5% of females having sex with more than one individual. However, of those arriving without sexual partners (75.5%) just under half (47.5%) have sex in Ibiza and most of these (62.4%) always used condoms. Having any sex abroad was associated with using illicit drugs and having more sexual partners in the 6 months before visiting Ibiza. However, having unprotected sex or sex with more than one person was associated with smoking as well as having higher numbers of sexual partners before their visit. Overall, 8.6% of individuals had sex with a non-UK resident in Ibiza although such individuals were no more likely to have sex without condoms. CONCLUSIONS: Substantial numbers of individuals visiting international nightlife resorts have unprotected sex with people they meet while abroad. This poses an increasing threat to the sexual health of UK residents but as yet little attention has been paid to developing interventions that might reduce sexual risk taking among young people holidaying abroad. PMID- 14755037 TI - Vulvovaginal candida in a young sexually active population: prevalence and association with oro-genital sex and frequent pain at intercourse. AB - OBJECTIVE: To study the prevalence of vulvovaginal candida among sexually active adolescents. To determine past and present symptoms, including pain at intercourse and potential behavioural risk factors associated with vulvovaginal candidiasis. METHODS: At an adolescent centre, 219 sexually active women who underwent genital examination, also completed a questionnaire on a history of genital symptoms and infections, sexual and hygiene habits, and the use of contraceptives. Symptoms and clinical signs were registered. Vaginal samples were analysed for candida species and urine for Chlamydia trachomatis. RESULTS: Candida culture was positive in 42% of the women and only 15% were asymptomatic. A history of recurrent candidiasis was given by 22%. Frequent pain at intercourse was reported by 24% and frequent oro-genital sex by 42% of the women. Frequent pain at intercourse was significantly associated with both the growth of candida and a history of recurrent candidiasis. Oro-genital sex was an independent risk factor for the growth of candida. CONCLUSION: In sexually active adolescents, who underwent genital examination, candida cultures were positive in 42%. The habit of frequent oro-genital sex was associated with the growth of candida. Pain at intercourse was associated with the growth of candida and recurrent candidiasis. PMID- 14755036 TI - Vaginal yeast colonisation, prevalence of vaginitis, and associated local immunity in adolescents. AB - OBJECTIVES: To evaluate point prevalence vaginal yeast colonisation and symptomatic vaginitis in middle adolescents and to identify relation of these yeast conditions with reproductive hormones, sexual activity, sexual behaviours, and associated local immunity. METHODS: Middle adolescent females (n = 153) were evaluated for sexually transmitted infections (STIs), asymptomatic yeast colonisation, and symptomatic vulvovaginal candidiasis (VVC) by standard criteria. Also evaluated were local parameters, including vaginal associated cytokines, chemokines, and antibodies, vaginal epithelial cell antifungal activity, and Candida specific peripheral blood lymphocyte responses. Correlations between yeast colonisation/vaginitis and local immunomodulators, reproductive hormones, douching, sexual activity, condom use, and STIs were identified. RESULTS: Rates of point prevalence asymptomatic yeast colonisation (22%) were similar to adults and similarly dominated by Candida albicans, but with uncharacteristically high vaginal yeast burden. In contrast with the high rate of STIs (18%), incidence of symptomatic VVC was low (<2%). Immunological properties included high rates of Candida specific systemic immune sensitisation, a Th2 type vaginal cytokine profile, total and Candida specific vaginal antibodies dominated by IgA, and moderate vaginal epithelial cell anti-Candida activity. Endogenous reproductive hormones were in low concentration. Sexual activity positively correlated with vaginal yeast colonisation, whereas vaginal cytokines (Th1, Th2, proinflammatory), chemokines, antibodies, contraception, douching, or condom use did not. CONCLUSION: Asymptomatic vaginal yeast colonisation in adolescents is distinct in some ways with adults, and positively correlates with sexual activity, but not with local immunomodulators or sexual behaviours. Despite several factors predictive for VVC, symptomatic VVC was low compared to STIs. PMID- 14755039 TI - Social and sexual risk factors for bacterial vaginosis. AB - BACKGROUND: A number of sexual and social risk factors for bacterial vaginosis (BV) have been identified. However, many previous studies have used small numbers of patients, or highly selected or convenience samples, or poorly defined populations. This study aims to clarify potential sexual and non-sexual risk factors for BV. METHODS: Women attending the Sydney Sexual Health Centre with BV, between March 1991 and July 1999, were included. Controls were randomly selected women without BV. Information on the demographics, clinical findings, and sexual and non-sexual risk behaviours were extracted from the clinic database and analysed using SPSS and SAS. A logistic regression model was used to establish which associations with BV persisted. RESULTS: 890 women with BV and 890 controls were studied. Factors that were independently associated with BV were > or =3 male sexual partners in the past 12 months (OR = 1.60, 95% CI: 1.19 to 2.04), at least one female sexual partner in the past 12 months (OR = 2.1, p = 0.003), a past pregnancy (OR = 1.5, p<0.0006), and smoking. In contrast, women with BV were significantly less likely to have used hormonal contraception (OR = 0.60, 95% CI: 0.51 to 0.81) or to have used condoms consistently (OR = 0.5, 95% CI: 0.31 to 0.71) than controls. CONCLUSION: Our findings may be important for planning a preventive strategy for BV by discouraging smoking and increasing condom use and hormonal contraception among women. PMID- 14755040 TI - Diagnosis of bacterial vaginosis: need for validation of microscopic image area used for scoring bacterial morphotypes. AB - BACKGROUND: The diagnosis of bacterial vaginosis (BV) is often made according to Nugent's classification, a scoring system based on bacterial counting of Gram stained slides of vaginal secretion. However as the image area of the microscope field will influence the number of morphotypes seen there is a need to standardise the area. METHODS: A graph intended for recalculation of number of bacterial morphotypes seen by the observer using 1000 x magnification from various microscope set-ups was constructed and applied to data sets typical for scoring BV. The graph was used in recalculation of Nugent scores, which were also compared with the Ison/Hay scores to evaluate the consequences for the diagnosis of BV. RESULTS: The observed image area differed by 300% among the investigated microscope set-ups. In two different data sets, one treatment study and one screening study, a considerable change in the number of women classified as intermediate was seen when the graph was used to standardise the image area. The recalculated numbers were also compared to the Ison/Hay classification. Weighted kappa indexes between the different methods were 0.84, 0.88, and 0.90, indicating that the methods are comparable. CONCLUSION: Because of the considerable differences among image areas covered by different microscope set-ups used in Nugent and Ison/Hay scoring, there is a need to standardise the area in order to reach comparable scores reflecting the diagnosis of BV in different laboratories. The differences in the intermediate group will have a considerable effect on the results from both treatment and prevalence studies, even though the kappa indexes indicate very good agreement between the methods used. PMID- 14755041 TI - Evaluation of real time polymerase chain reaction assays for confirmation of Neisseria gonorrhoeae in clinical samples tested positive in the Roche Cobas Amplicor assay. AB - OBJECTIVE: Development of a rapid, sensitive, and accurate assay for confirmation of Neisseria gonorrhoeae in clinical samples. METHOD: Two real time polymerase chain reaction (PCR) assays, developed on the LightCycler for amplification of the N gonorrhoeae cppB gene, were utilised for confirmation of this bacterial pathogen in samples positive by the Roche Cobas Amplicor assay. Performance characteristics of the two assays were compared with other commercial nucleic acid amplification assays, including the Abbott LCx and Roche 16S rRNA tests. RESULTS: All related Neisseria as well as other bacterial species tested negative by both cppB gene based assays, whereas 120 N gonorrhoeae clinical isolates from various geographical regions gave in positive results. Both assays had a sensitivity of one copy per reaction. 122 clinical samples positive and another 50 samples negative for N gonorrhoeae by Roche Cobas Amplicor were selected from a specimen pool of more than 3000 women tested previously. Overall, 73 of 122 (59.8%) samples were confirmed as positive. The two real time assays had sensitivities of 99% and 100% and specificities of 98% and 100%, respectively. The 16S and LCx assays produced similar results to the real time assays, indicating a similar sensitivity to and specificity of both real time assays. CONCLUSION: The data from this study highlight the need to confirm N gonorrhoeae positive Cobas Amplicor PCR results as an important part of the testing algorithm of all diagnostic laboratories utilising this assay. PMID- 14755042 TI - Localised genital Norwegian scabies in an AIDS patient. AB - OBJECTIVES: We present a case of an AIDS patient with Norwegian scabies manifest by a single, crusted plaque localised to the glans penis. METHODS: A 45 year old man with AIDS presented to our clinic complaining of a red papular pruritic rash on his abdomen and anterior thighs and a single, thick, crusted, non-pruritic lesion on the penis. He had been treated with lindane topically prior to the development of the penile lesion without resolution of the pruritus or red papular lesions. A mineral oil preparation was obtained from the hyperkeratotic penile lesion and revealed numerous mite eggs and faeces. RESULTS: The diagnosis of localised, genital Norwegian scabies was made. The patient was treated with ivermectin 200 micro g/kg per dose taken as two doses, 14 days apart, with complete resolution of both pruritus and skin lesions. CONCLUSIONS: This patient is the first known report of Norwegian scabies localised as a single lesion on the penis. He was successfully treated with oral ivermectin monotherapy. PMID- 14755043 TI - A new method for extended trichomonad storage. PMID- 14755044 TI - Perceived transmissibility of STIs: lack of differentiation between HIV and chlamydia. PMID- 14755045 TI - South Asians with HIV in London: is it time to rethink sexual health service delivery to meet the needs of heterosexual ethnic minorities? PMID- 14755046 TI - "Water can" penis caused by tuberculosis. PMID- 14755047 TI - Failure to maintain patient access to GUM clinics. PMID- 14755048 TI - Prevalence of HSV-1/HSV-2 antibodies in HIV seropositive patients in Coventry, United Kingdom. PMID- 14755051 TI - Imaging reversal of multidrug resistance in living mice with bioluminescence: MDR1 P-glycoprotein transports coelenterazine. AB - Coelenterazine is widely distributed among marine organisms, producing bioluminescence by calcium-insensitive oxidation mediated by Renilla luciferase (Rluc) and calcium-dependent oxidation mediated by the photoprotein aequorin. Despite its abundance in nature and wide use of both proteins as reporters of gene expression and signal transduction, little is known about mechanisms of coelenterazine transport and cell permeation. Interestingly, coelenterazine analogues share structural and physiochemical properties of compounds transported by the multidrug resistance MDR1 P-glycoprotein (Pgp). Herein, we report that living cells stably transfected with a codon-humanized Rluc show coelenterazine mediated bioluminescence in a highly MDR1 Pgp-modulated manner. In Pgp-expressing Rluc cells, low baseline bioluminescence could be fully enhanced (reversed) to non-Pgp matched control levels with potent and selective Pgp inhibitors. Therefore, using coelenterazine and noninvasive bioluminescence imaging in vivo, we could directly monitor tumor-specific Pgp transport inhibition in living mice. While enabling molecular imaging and high-throughput screening of drug resistance pathways, these data also raise concern for the indiscriminate use of Rluc and aequorin as reporters in intact cells or transgenic animals, wherein Pgp-mediated alterations in coelenterazine permeability may impact results. PMID- 14755052 TI - Histone H3 lysine 9 methylation is required for DNA elimination in developing macronuclei in Tetrahymena. AB - Genome-wide DNA elimination accompanies development of the somatic macronucleus from the germ-line micronucleus during the sexual process of conjugation in the ciliated protozoan Tetrahymena thermophila. Small RNAs, referred to as "scan RNAs" (scnRNAs), that accumulate only during conjugation are highly enriched in the eliminated sequences, and mutations that prevent DNA elimination also affect the accumulation of scnRNAs, suggesting that an RNA interference (RNAi)-like mechanism is involved in DNA elimination. Histone H3 that is methylated at lysine 9 (K9) is a hallmark of heterochromatin and, in Tetrahymena, is found only in developing macronuclei (anlagen) in association with chromatin containing the sequences undergoing elimination. In this article, we demonstrate that a mutation in the TWI1 gene that eliminates the accumulation of scnRNAs also abolishes H3 methylation at K9. We created mutant strains of Tetrahymena in which the only major H3 contained a K9Q mutation. These mutants accumulated scnRNAs normally during conjugation but showed dramatically reduced efficiency of DNA elimination. These results provide strong genetic evidence linking an RNAi-like pathway, H3 K9 methylation, and DNA elimination in Tetrahymena. PMID- 14755053 TI - Exceptionally fast self-cleavage by a Neurospora Varkud satellite ribozyme. AB - Most of the small ribozymes, including those that have been investigated as potential therapeutic agents, appear to be rather poor catalysts. These RNAs use an internal phosphoester transfer mechanism to catalyze site-specific RNA cleavage with apparent cleavage rate constants typically <2 min(-1). We have identified variants of one of these, the Neurospora Varkud satellite ribozyme, that self-cleaves with experimentally measured apparent rate constants of up to 10 s(-1) (600 min(-1)), approximately 2 orders of magnitude faster than any previously characterized self-cleaving RNA. We describe structural features of the cleavage site loop and an adjacent helix that affect the apparent rate constants for cleavage and ligation and the equilibrium between them. These data show that the phosphoester transfer ribozymes can catalyze reactions with rate constants much larger than previously appreciated and in the range of those of protein enzymes that perform similar reactions. PMID- 14755054 TI - Glucose sensing and signaling in Saccharomyces cerevisiae through the Rgt2 glucose sensor and casein kinase I. AB - The yeast Saccharomyces cerevisiae senses glucose through two transmembrane glucose sensors, Snf3 and Rgt2. Extracellular glucose causes these sensors to generate an intracellular signal that induces expression of HXT genes encoding glucose transporters by inhibiting the function of Rgt1, a transcriptional repressor of HXT genes. We present the following evidence that suggests that the glucose sensors are coupled to the membrane-associated protein kinase casein kinase I (Yck1). (i) Overexpression of Yck1 leads to constitutive HXT1 expression; (ii) Yck1 (or its paralogue Yck2) is required for glucose induction of HXT1 expression; (iii) Yck1 interacts with the Rgt2 glucose sensor; and (iv) attaching the C-terminal cytoplasmic tail of Rgt2 to Yck1 results in a constitutive glucose signal. The likely targets of Yck1 in this signal transduction pathway are Mth1 and Std1, which bind to and regulate function of the Rgt1 transcription factor and bind to the C-terminal cytoplasmic domain of glucose sensors. Potential casein kinase I phosphorylation sites in Mth1 and Std1 are required for normal glucose regulation of HXT1 expression, and Yck1 catalyzes phosphorylation of Mth1 and Std1 in vitro. These results support a model of glucose signaling in which glucose binding to the glucose sensors causes them to activate Yck1 in the cell membrane, which then phosphorylates Mth1 and Std1 bound to the cytoplasmic face of the glucose sensors, triggering their degradation and leading to the derepression of HXT gene expression. Our results add nutrient sensing to the growing list of processes in which casein kinase I is involved. PMID- 14755055 TI - In vitro synthesis of fully functional EmrE, a multidrug transporter, and study of its oligomeric state. AB - EmrE is a small multidrug transporter from Escherichia coli that provides a unique model for the study of polytopic membrane proteins. Here, we show its synthesis in a cell-free system in a fully functional form. The detergent solubilized protein binds substrates with high affinity and, when reconstituted into proteoliposomes, transports substrate in a Deltamicro(H)(+)-dependent fashion. Here, we used the cell-free system to study the oligomeric properties of EmrE. EmrE functions as an oligomer, but the size of the functional oligomer has not been established unequivocally. Coexpression of two plasmids in the cell-free system allowed demonstration of functional complementation and pull-down experiments confirmed that the basic functional unit is the dimer. An additional interaction between dimers has been detected by using crosslinking between unique Cys residues. This finding implies the existence of a dimer of dimers. PMID- 14755056 TI - Increased activity of antagonists of growth hormone-releasing hormone substituted at positions 8, 9, and 10. AB - Antagonists of human growth hormone-releasing hormone (hGHRH) with increased potency and improved enzymatic and chemical stability are needed for potential clinical applications. We synthesized 21 antagonistic analogs of hGHRH(1 29)NH(2), substituted at positions 8, 9, and 10 of the common core sequence [phenylacetyl-Tyr(1), d-Arg(2,28), para-chloro-phenylalanine 6, Arg(9)/homoarginine 9, Tyr(10)/O-methyltyrosine 10, alpha-aminobutyric acid 15, norleucine 27, Har(29)] hGHRH(1-29)NH(2). Inhibitory effects on hGHRH-induced GH release were evaluated in vitro in a superfused rat pituitary system, as well as in vivo after i.v. injection into rats. The binding affinities of the peptides to pituitary GHRH receptors were also determined. Introduction of para amidinophenylalanine 10 yielded antagonists JV-1-62 and -63 with the highest activities in vitro and lowest receptor dissociation constants (K(i) = 0.057 0.062 nM). Antagonists JV-1-62 and -63 also exhibited the strongest effect in vivo, significantly (P < 0.05-0.001) inhibiting hGHRH-induced GH release for at least 1 h. Para-aminophenylalanine 10 and O-ethyltyrosine 10 substitutions yielded antagonists potent in vitro, but His(10), 3,3'-diphenylalanine 10, 2 naphthylalanine 10, and cyclohexylalanine 10 modifications were detrimental. Antagonists containing citrulline 9 (in MZ-J-7-72), amidinophenylalanine 9 (in JV 1-65), His(9), d-Arg(9), citrulline 8, Ala(8), d-Ala(8), or alpha-aminobutyric acid 8 substituents also had high activity and receptor affinity in vitro. However, in vitro potencies of analogs with substitution in position 9 correlated poorly with acute endocrine effects in vivo, as exemplified by the weak and/or short inhibitory actions of antagonists JV-1-65 and MZ-J-7-72 on GH release in vivo. Nevertheless, antagonist JV-1-65 was more potent than JV-1-63 in tests on inhibition of the growth of human prostatic and lung cancer lines xenografted into nude mice. This indicates that oncological activity may be based on several mechanisms. hGHRH antagonists with improved efficacy could be useful for treatment of cancers that depend on insulin-like growth factors or GHRH. PMID- 14755057 TI - STAT1 is overexpressed in tumors selected for radioresistance and confers protection from radiation in transduced sensitive cells. AB - Nu61, a radiation-resistant human tumor xenograft, was selected from a parental radiosensitive tumor SCC-61 by eight serial cycles of passage in athymic nude mice and in vivo irradiation. Replicate DNA array experiments identified 52 genes differentially expressed in nu61 tumors compared with SCC-61 tumors. Of these, 19 genes were in the IFN-signaling pathway and moreover, 25 of the 52 genes were inducible by IFN in the nu61 cell line. Among the genes involved in IFN signaling, STAT1alpha and STAT1beta were the most highly overexpressed in nu61 compared to SCC-61. STAT1alpha and STAT1beta cDNAs were cloned and stably transfected into SCC-61 tumor cells. Clones of SCC-61 tumor cells transfected with vectors expressing STAT1alpha and STAT1beta demonstrated radioprotection after exposure to 3 Gy (P < 0.038). The results indicate that radioresistance acquired during radiotherapy treatment may account for some treatment failures and demonstrate an association of acquired tumor radioresistance with up regulation of components of the IFN-related signaling pathway. PMID- 14755058 TI - The gene for soluble N-ethylmaleimide sensitive factor attachment protein alpha is mutated in hydrocephaly with hop gait (hyh) mice. AB - The spontaneous autosomal recessive mouse mutant for hydrocephaly with hop gait (hyh) exhibits dramatic cystic dilation of the ventricles at birth and invariably develops hopping gait. We show that the gene for soluble N-ethylmaleimide sensitive factor attachment protein alpha, also known as alpha-SNAP, is mutated in hyh mice. alpha-SNAP plays a key role in a wide variety of membrane fusion events in eukaryotic cells, including the regulated exocytosis of neurotransmitters. Homozygous mutant mice harbor a missense mutation M105I in a conserved residue in one of the alpha-helical domains. We demonstrate that the hyh mutant is not a null allele and is expressed; however, the mutant protein is 40% less abundant in hyh mice. The hyh mutant provides a valuable in vivo model to study vesicle/membrane trafficking and provides insight into the potential roles of alpha-SNAP in embryogenesis and brain development. PMID- 14755059 TI - Cell-cell signaling controls Xylella fastidiosa interactions with both insects and plants. AB - Xylella fastidiosa, which causes Pierce's disease of grapevine and other important plant diseases, is a xylem-limited bacterium that depends on insect vectors for transmission. Although many studies have addressed disease symptom development and transmission of the pathogen by vectors, little is known about the bacterial mechanisms driving these processes. Recently available X. fastidiosa genomic sequences and molecular tools have provided new routes for investigation. Here, we show that a diffusible signal molecule is required for biofilm formation in the vector and for vector transmission to plants. We constructed strains of X. fastidiosa mutated in the rpfF gene and determined that they are unable to produce the signal activity. In addition, rpfF mutants are more virulent than the wild type when mechanically inoculated into plants. This signal therefore directs interaction of X. fastidiosa with both its insect vector and plant host. Interestingly, rpfF mutants can still form in planta biofilms, which differ architecturally from biofilms in insects, suggesting that biofilm architecture, rather than a passive response to the environment, is actively determined by X. fastidiosa gene expression. This article reports a cell-cell signaling requirement for vector transmission. Identification of the genes regulated by rpfF should elucidate bacterial factors involved in transmission and biofilm formation in the insect. PMID- 14755060 TI - Molecular dissection of the roles of nucleotide binding and hydrolysis in dynein's AAA domains in Saccharomyces cerevisiae. AB - The motor protein cytoplasmic dynein is responsible for most of the minus-end directed microtubule traffic within cells. Dynein contains four evolutionarily conserved AAA (ATPase associated with various cellular activities) domains that are thought to bind nucleotide; the role of nucleotide binding and hydrolysis in each of these four AAA domains has constituted an important and unresolved question in understanding dynein's mechanism. Using Saccharomyces cerevisiae cytoplasmic dynein as a model system, we mutagenized residues involved in nucleotide binding or hydrolysis in the four AAA domains and examined the ability of the mutant dyneins to mediate nuclear segregation in vivo and to bind microtubules in vitro. Our analysis shows that an AAA1 hydrolysis mutant blocks dynein function, whereas a triple AAA2/3/4 hydrolysis mutant does not, suggesting that nucleotide binding is required at only one site. We also show that nucleotide binding at AAA3, but not hydrolysis, is essential for motor activity in vivo and ATP-induced dissociation of dynein from microtubules, suggesting that this domain acts as a critical allosteric site. In contrast, mutations in AAA2 cause subtle defects in dynein function, whereas mutation in AAA4 produce no obvious defects. These results show that the four conserved dynein AAA domains have distinct functions in dynein's mechanochemical cycle. PMID- 14755061 TI - Comment: Factors affecting patient adherence to highly active antiretroviral therapy. PMID- 14755063 TI - Compounded versus proprietary sincalide for evaluation of gallbladder ejection fraction. AB - OBJECTIVE: To report a comparison of compounded and proprietary sincalide in the evaluation of gallbladder ejection fraction during hepatobiliary scintigraphy. CASE SUMMARIES: Two patients were referred to nuclear medicine with symptoms consistent with hepatobiliary dysfunction. Both underwent hepatobiliary scintigraphy to evaluate anatomic and physiologic tract patency of the hepatobiliary system. Compounded sincalide, an adjuvant pharmaceutical used to evaluate gallbladder ejection fraction, was infused during hepatobiliary scintigraphy, and gallbladder ejection fractions were 11% and 24%, respectively. Hepatobiliary scintigraphy was repeated on both patients 72 hours later with proprietary sincalide used as the adjuvant pharmaceutical. The gallbladder ejection fractions were 32% and 72%, respectively. DISCUSSION: The use of sincalide to evaluate gallbladder ejection fraction in hepatobiliary scintigraphy is widely accepted in the surgical and nuclear medicine community. In late 2001, the sole manufacturer of sincalide announced indefinite unavailability of the product. Following the announcement, several compounding pharmacies began selling extemporaneously compounded sincalide as a replacement. Use of the compounded product has assumed therapeutic equivalence. CONCLUSIONS: Significant differences in gallbladder ejection fraction between compounded sincalide and sincalide in our patients are likely due to the intrinsic variability in response to sincalide. Clinicians should be aware of this variability, as well as the potential effect of concomitant medications. PMID- 14755064 TI - Pharmacologic therapy for HIV-associated lipodystrophy. AB - OBJECTIVE: To evaluate the efficacy and safety of pharmacologic therapy in the treatment of HIV-associated lipodystrophy, with a focus on the treatment of fat redistribution. Drug therapies that have been shown to be beneficial in other forms of lipodystrophy and are currently being evaluated in HIV-associated lipodystrophy are also discussed. DATA SOURCES: A MEDLINE search was conducted from 1996 to February 2003. Bibliographies of all articles were reviewed and pertinent articles were included. Abstracts from major meetings in 2002 and 2003 were also reviewed. STUDY SELECTION AND DATA EXTRACTION: All published studies were included in the review. DATA SYNTHESIS: Lipodystrophy has become more prevalent in patients with HIV. Lipodystrophy consists of adipose redistribution and metabolic abnormalities including dyslipidemia and insulin resistance. Treatment of lipodystrophy has been directed at either decreasing the amount of visceral adipose tissue (VAT), dorsocervical adipose tissue (commonly known as buffalo hump) and/or increase subcutaneous adipose tissue (SAT). Recombinant human growth hormone (rhGH) decreases VAT and buffalo hump, although it has been associated with a high frequency of adverse effects. Metformin and the thiazolidinediones have favorable metabolic effects, but were not found to be effective in correcting body compositional changes associated with lipodystrophy. Anabolic steroids and l-carnitine are not effective in the treatment of lipodystrophy. CONCLUSIONS: No drug therapy exists to fully ameliorate or correct the cosmetic changes of HIV-associated lipodystrophy. Clinicians must weigh the benefits and risks of each agent and individualize treatment for each patient. PMID- 14755065 TI - Voriconazole in fungal keratitis caused by Scedosporium apiospermum. AB - OBJECTIVE: To describe the first case, to the best of our knowledge, of posttraumatic Scedosporium apiospermum (ScA) keratitis successfully treated with systemic and topical voriconazole. CASE SUMMARY: A 19-year-old man was admitted to the hospital with an incisive wound of his left eye and the cornea totally sectioned after trauma with a cutter used in gardening. Initial empirical treatment was followed by systemic and topical voriconazole, and the eye did not have to be enucleated. Five months after the trauma, a penetrating keratoplasty and chamber intraocular lens implantation was performed with a favorable visual outcome. DISCUSSION: ScA keratitis is rare, but it must be suspected if a history of ocular injury with contaminated objects exists. Among the antifungals available to treat ScA keratitis, voriconazole has shown advantages such as the lowest minimum inhibitory concentration and the availability of an oral formulation. CONCLUSIONS: Voriconazole shows promise as an effective alternative to conventional antifungals in the treatment of ScA keratitis. It is available both as oral and intravenous preparations, which is a great advantage in these lengthy infections. PMID- 14755066 TI - Smallpox: clinical features, prevention, and management. AB - OBJECTIVE: To describe a general overview of smallpox, clinical presentation, diagnosis, adverse events, and management of both pre- and postexposure vaccination. DATA SOURCES: Literature was identified by search of MEDLINE (1966 June 2003) and International Pharmaceutical Abstracts (1966-May 2003) databases using the key terms smallpox, bioterrorism, biological warfare, and smallpox vaccine. STUDY SELECTION AND DATA EXTRACTION: Articles identified from data sources were evaluated, and relevant information was included in this review. DATA SYNTHESIS: Smallpox is spread by human-to-human contact with an infected host and therefore can be contagious. The mortality rate for smallpox is approximately 30%. While the disease was completely eradicated by 1980 with successful use of smallpox vaccine, concern has been raised that smallpox may emerge as a tool of bioterrorism. This concern, combined with the reality of current smallpox vaccination programs in the military and selected civilian populations, mandates a clear understanding of vaccination-related adverse events and contraindications by all healthcare professionals. The vaccine may cause moderate to severe adverse events such as eczema vaccinatum, progressive vaccinia, and generalized vaccinia. CONCLUSIONS: The balance between the risks and benefits of mass vaccination in prevention of an epidemic is not clear. The Centers for Disease Control and Prevention has established a guideline for appropriate use of smallpox vaccine in the civilian population. PMID- 14755067 TI - Acute pancreatitis and modified-release clarithromycin. PMID- 14755068 TI - Complementary medicine prescription patterns in Germany. AB - BACKGROUND: There is growing interest in complementary medicine worldwide and a corresponding need to know how patients and practitioners interact. OBJECTIVE: To chart the use of complementary medications compared with conventional treatments among practitioners and patients in Germany. METHODS: Comparative analysis of patients' data from a prospective cohort study of 4178 patients presenting with chronic as well as acute symptoms by 218 practitioners at 218 centers in all German federal states between 2001 and 2002. Practices focused on either conventional medicine, complementary medicine, or both. Data were gathered on prescription patterns, treatment satisfaction and adherence, and the degree of patient involvement in treatment decisions. RESULTS: Complementary medicine was preferentially prescribed over conventional medicine in patients <18 and >65 years old, in women, and in patients with chronic symptoms. Patients receiving complementary medications had, on average, higher rates of accompanying illnesses, received more preparations, and were more closely involved in the decision process than patients prescribed conventional therapies. The decision to use complementary medicine was based on both the patient's and the practitioner's wishes in 40.8% of the cases compared with 25.8% of the cases of conventional therapies. Tolerability and satisfaction with treatment appeared greater with complementary than with conventional therapies. CONCLUSIONS: Complementary medicine is generally well established in Germany, apparently in a dialogue with conventional medicine. Patients receiving complementary medicine appear to be more closely involved in the decision process and more satisfied with treatments than conventionally treated patients. PMID- 14755070 TI - Parasites and allergy: from IgE to Th1/Th2 and beyond. PMID- 14755071 TI - The potential impact of early exposures to geohelminth infections on the development of atopy. AB - Microbial exposures in early life may provide important signals for immune maturation and the development of an antiinflammatory network thereby preventing the development of dysregulated immune responses such as that associated with allergic disease. The human immune system has evolved in the presence of intense helminth infections and has developed regulatory mechanisms to limit the harmful inflammation that can be caused by the potent allergens secreted by these chronic pathogens. Geohelminth infections are highly prevalent childhood infections, and there is strong evidence that chronic geohelminth infections provide protection against atopy in the rural tropics. Because the early environmental exposures that may lead to the development of atopy are likely to occur in the first few years of life, geohelminth infections may exert their protective effects at this time. Early exposures to geohelminth antigens could occur transplacentally, through breast milk, or through early infant exposures, and could induce tolerance to parasite antigens resulting in suppressed allergic responses to the parasite. Tolerization to parasite antigens could suppress allergic responses to inhalant allergens through bystander effects or through tolerization of crossreactive epitopes that are shared between geohelminth parasites and inhalant allergens. Tolerization to crossreactive allergens could occur by thymic deletion or through peripheral mechanisms such as regulatory T cells. Immunologic studies of the mechanisms by which early exposures to geohelminth infections affect immune polarization to inhalant allergens are likely to provide important insights into the early regulation of the immune response and may lead to the design of novel interventions for the prevention of allergic disease. PMID- 14755072 TI - Parasites and the hygiene hypothesis: regulating the immune system? AB - The hygiene hypothesis proposes that the stimulation of the immune system by microbes or microbial products protects from the development of inflammatory diseases; therefore a reduced exposure to infectious agents may explain the rise in allergic and autoimmune diseases in industrialized countries. The contribution of studies on parasites and allergy to our understanding of the hygiene hypothesis has been two-fold. First, several studies have shown an inverse association between exposure to (Toxplasma gondii) or harbouring of parasites (Schistosoma or Intestinal helminths) and allergy. Second, the mechanisms behind such protective effects have provided new insights and theories on the ability of parasite derived molecules to down-regulate immune responses and thereby to control inflammatory diseases such as allergies. In this review, recent findings and new concepts relating to the associations between parasites and allergies at the epidemiological, cellular and molecular level are discussed. PMID- 14755073 TI - Helminth infections and allergic diseases: from the Th2 paradigm to regulatory networks. AB - Both helminth infections and allergic diseases are characterized by a Th2 response. The former mostly affects developing countries while the later is steadily increasing in westernized countries, since most parasitic diseases have been eradicated. However, besides the cytokine pattern, both pathologies display striking similarities regarding the cell types involved in the process, the mucosal component of the inflammatory reaction and the immunoglobulin isotypes produced, IgE and IgA. Recent advances also suggest that allergic pathologies and some other inflammatory diseases arise from a similar dysfunction of the regulatory mechanisms, existing within the immune system. Such a dysregulation leads to a rupture of tolerance towards allergen. Chronic and moderate helminth infections might be beneficial to the induction of various regulatory mechanisms, involving specific regulatory cell populations (dendritic cells and lymphocytes), inhibitory receptors, blocking antibodies and two prominent cytokines: IL-10 and TGF-b, which would decrease the incidence of allergic diseases. This mechanism could provide a new vision of the so-called "hygiene hypothesis". PMID- 14755077 TI - The ERK cascade as a prototype of MAPK signaling pathways. PMID- 14755078 TI - Determination of ERK activity: antiphospho-ERK antibodies, in vitro phosphorylation, and in-gel kinase assay. PMID- 14755076 TI - The innate allergenicity of helminth parasites. AB - Helminth parasites are well known to induce an immune response in their hosts characterised by elevated IgE, peripheral blood or local tissue eosinophilia, and in some cases, intestinal mastocytosis. This immunological response has a strong T-helper 2 (Th2) cytokine bias and is reminiscent of the immunological constellation found in allergic diseases. However, the molecular forces driving the Th2 response to helminth parasites are still not understood. By using the human hookworm parasite Necator americanus as an example, the authors of the current article propose that in the course of its life cycle, this parasite becomes innately allergenic through the secretion of a molecular array designed to promote tissue migration and homing, feeding and survival against immunological attack. This complex array comprises proteases, lectins and other classes of molecules. Subsequent immunological and physiological events seemingly protect the host from both the allergic sequelae of exposure to environmental allergens and, moreover, from the parasite itself. PMID- 14755074 TI - Regulation of allergy and autoimmunity in helminth infection. AB - Parasitic infections are a major theme in the "hygiene hypothesis", as allergies and autoimmune diseases are less prevalent in countries with higher burdens of helminths and other parasitic organisms. Helminths"-the grouping of multicellular worm parasites including nematodes, cestodes and trematodes-tend to establish long-lived, chronic infections indicating successful down-modulation of the host immune system. In this review, we describe the intricate immunology of host helminth interactions and how parasites manipulate immune responses to enhance their survival. In so doing, they often minimise immunopathology and, it is suggested, reduce host susceptibility to, and severity of allergic and autoimmune diseases. Studies on helminth-infected communities and individuals support the hypothesis that an immuno-regulatory network promoted by parasites extends its influence to limiting allergies. Experimental models are now probing more deeply into the area of immune modulation by helminths, and we discuss the likely mechanisms by which helminths could be establishing a strongly regulatory environment. Understanding and harnessing the modulatory capacity of helminths may uncover novel therapeutic interventions, mimicking and exploiting their evolution for our benefit. Parasitic infections are a major theme in the "hygiene hypothesis", as allergies and autoimmune diseases are less prevalent in countries with higher burdens of helminths and other parasitic organisms. Helminths"-the grouping of multicellular worm parasites including nematodes, cestodes and trematodes-tend to establish long-lived, chronic infections indicating successful down-modulation of the host immune system. In this review, we describe the intricate immunology of host-helminth interactions and how parasites manipulate immune responses to enhance their survival. In so doing, they often minimise immunopathology and, it is suggested, reduce host susceptibility to, and severity of allergic and autoimmune diseases. Studies on helminth-infected communities and individuals support the hypothesis that an immuno-regulatory network promoted by parasites extends its influence to limiting allergies. Experimental models are now probing more deeply into the area of immune modulation by helminths, and we discuss the likely mechanisms by which helminths could be establishing a strongly regulatory environment. Understanding and harnessing the modulatory capacity of helminths may uncover novel therapeutic interventions, mimicking and exploiting their evolution for our benefit. PMID- 14755075 TI - Interleukin-13: a key mediator in resistance to gastrointestinal-dwelling nematode parasites. AB - Gastrointestinal nematode parasites are one of the most prevalent types of infection worldwide. Evidence from both laboratory and human systems indicates that when resistance is evident immunity is mediated by effector mechanisms controlled by T helper 2 type responses. Moreover, more recent evidence implicates a central role for interleukin 13. We raise the possibility that gut dwelling nematodes may have been an important driving force in the development of Th 2 responses involving IL-13. Moreover, that these parasites have evolved a variety of strategies to avoid destruction and to regulate any potential pathology associated with chronic infection. PMID- 14755079 TI - Detection of ERK1/2 activities using affinity reagents. PMID- 14755081 TI - Investigating the cellular BMK1/ERK5 signaling pathway. PMID- 14755080 TI - Activation of SAPKs/JNKs and p38s in vitro. PMID- 14755082 TI - Pull-down assays for guanoside 5'-triphosphate-bound Ras-like guanosine 5' triphosphatases. PMID- 14755083 TI - Regulation of MAPK cascades by protein tyrosine phosphatases. PMID- 14755084 TI - Use of inhibitors in the study of MAPK signaling. PMID- 14755085 TI - Structure of MAPKs. PMID- 14755086 TI - Subcellular localization of MAPKs. PMID- 14755087 TI - Study of MAPK signaling using knockout mice. PMID- 14755088 TI - Computer simulation of MAPK signal transduction. PMID- 14755089 TI - Signaling by growth factor receptors. PMID- 14755090 TI - Activation of MAPKs by G protein-coupled receptors. PMID- 14755091 TI - Identification of MAPK substrates by expression screening with solid-phase phosphorylation. PMID- 14755092 TI - Study of substrate specificity of MAPKs using oriented peptide libraries. PMID- 14755093 TI - Ras signaling pathway for analysis of protein-protein interactions in yeast and mammalian cells. PMID- 14755094 TI - Methods in functional proteomics: two-dimensional polyacrylamide gel electrophoresis with immobilized pH gradients, in-gel digestion and identification of proteins by mass spectrometry. PMID- 14755095 TI - Practical methods for deuterium exchange/mass spectrometry. PMID- 14755096 TI - Use of Xenopus oocytes and early embryos to study MAPK signaling. PMID- 14755097 TI - MAPK cascades in the brain: lessons from learning. PMID- 14755098 TI - Adenosine receptors and wound healing. AB - Recent studies have demonstrated that application of topical adenosine A2A receptor agonists promotes more rapid wound closure and clinical studies are currently underway to determine the utility of topical A2A adenosine receptor agonists in the therapy of diabetic foot ulcers. The effects of adenosine A2A receptors on the cells and tissues of healing wounds have only recently been explored. We review here the known effects of adenosine A2A receptor occupancy on the cells involved in wound healing. PMID- 14755099 TI - The use of plants for remediation of metal-contaminated soils. AB - The use of green plants to remove, contain, inactivate, or degrade harmful environmental contaminants (generally termed phytoremediation) is an emerging technology. In this paper, an overview is given of existing information concerning the use of plants for the remediation of metal-contaminated soils. Both site decontamination (phytoextraction) and stabilization techniques (phytostabilization) are described. In addition to the plant itself, the use of soil amendments for mobilization (in case of phytoextraction) and immobilization (in case of phytostabilization) is discussed. Also, the economical impacts of changed land-use, eventual valorization of biomass, and cost-benefit aspects of phytoremediation are treated. In spite of the growing public and commercial interest and success, more fundamental research is needed still to better exploit the metabolic diversity of the plants themselves, but also to better understand the complex interactions between metals, soil, plant roots, and micro-organisms (bacteria and mycorrhiza) in the rhizosphere. Further, more demonstration experiments are needed to measure the underlying economics, for public acceptance and last but not least, to convince policy makers. PMID- 14755100 TI - Recurrence prevention in patients with urinary tract stone disease. AB - Formation of urinary tract concrements is a common disease and steps should be taken in order to elucidate the underlying mechanisms and to give the patients appropriate advice and medical treatment. This present article summarizes the principles for recurrence preventive measures in patients with uric acid, infection, cystine and calcium stone disease. Categories of stone formers are identified with the aim of providing a basis for an individualised treatment with a reasonable patient's compliance. The recommendations are in line with those given by the EAU guideline group for urolithiasis. PMID- 14755107 TI - Five theories of the human existence. PMID- 14755108 TI - The life mission theory II. The structure of the life purpose and the ego. AB - Pursuing your life mission is often very difficult, and many frustrations are experienced along the way. Major failures to bring out our potential can cause us considerable emotional pain. When this pain is unbearable, we are induced to shift from one intention and talent to another that better allows us to adapt and survive. Thus, we become set on a course that brings out a secondary or tertiary talent instead of the primary talent. This talent displacement may be expressed as a loss of our true nature or true self. The new purpose in life now functions as the core of a new personality: the ego. The ego has a structure similar to that of the true self. It is anchored in a talent and it draws on subtalents. But the person who is centered in his or her ego is not as powerful or talented as the person he or she originally was, living the primary purpose of life. This is because the original personality (the true self or "higher self") is still there, active and alive, behind the ego. Symptoms, disorders, and diseases may be explained by the loss of energy, joy in life, and intuitive competence because of inner conflicts, which may be alleviated or cured in the salutogenetic process of Antonovsky that helps patients find their sense of coherence or their primary purpose in life. Many cases of reduced ability to function, physically as well as psychologically, socially or sexually, can also be explained and alleviated in this way. When a person discovers his true talent and begins to use it with dedication, privately as well as professionally, his life will flourish and he may overcome even serious disease and great adversity in life. The salutogenetic process can also be called personal development or "quality of life as medicine". It is important to note that the plan for personal development laid out by this theory is a plan not for the elimination of the ego, but for its cultivation. An existentially sound person still has a mental ego of course, but it is centered on the optimal verbal expression of the life mission. Such an ego is not in conflict with one's true self, but supports the life and wholeness of the person, although in an invisible and seamless way. The more developed the person, the more talents are taken into use. So although the core of existence remains the same throughout life, the healthy person continues to grow. As the number of talents we can call on is unlimited, the journey ends only at death. Understanding the concept of the ego, it is very easy for the physician to motivate the patient to go through a lot of difficulties in order to grow and develop, and when the patient fully understands the concept of the ego and the true self (higher self), the patient gets a strong feeling of direction in personal development, and a motivation to fight the internal obstacles for quality of life, health, and the ability to function. PMID- 14755109 TI - The life mission theory III. Theory of talent. AB - When we acknowledge our purpose as the essence of our self, when we take all our power into use in an effortless way, and when we fully accept our own nature- including sex and sexuality, our purpose of life takes the form of a unique talent. Using this talent gives the experience of happiness. A person in his natural state of being uses his core talent in a conscious, joyful, and effortless way, contributing to the world the best he or she has to offer. Full expression of self happens when a person, in full acceptance of body and life, with whole-hearted intension, uses all his personal powers to realize his core talent and all associated talents, to contribute to his beloved and to the world. Thus, self-actualisation is a result of a person fully expressing and realizing his core talent. The theory of talent states that a core talent can be expressed optimally when a human being takes possession of a three-dimensional space with the axis of purpose, power and gender, as we have a threefold need: Acknowledging our core talent (our purpose of life) and intending it; Understanding our potential powers and manifesting them; Accepting our human form including our sex and expressing it. The first dimension is spiritual, the next dimension is mental, emotional and physical, and the third dimension is bodily and sexual. We manifest our talents in a giving movement from the bottom of our soul trough our biological nature onto the subject and object of the outer world. These three dimensions can be drawn as three axes, one saggital axis called purpose or love or me-you, one vertical axis called power or consciousness (light) or heaven earth, and one horizontal axis called gender or joy or male-female. The three core dimensions of human existence are considered of equal importance for expression of our life purpose, life mission, or core talent. Each of the dimensions is connected to special needs. When these needs are not fulfilled, we suffer and if this suffering becomes unbearable we deny the dimension or a part of is. This is why the dimensions of purpose, power and gender become suppressed from our consciousness. PMID- 14755110 TI - The life mission theory IV. Theory on child development. AB - We can identify five important needs that children have: the need for acknowledgment, acceptance, awareness or attention, respect, and care. If these needs are not met, children will modify themselves by denying central parts of their nature in order to adjust to their parents and the situation at large. When a child denies his or her talents, powers, and gender or aspects thereof, he or she loses quality of life, the ability to function, and physical or mental health. The loss of ability takes the form of diminished social ability, psychosexual potency, joy, energy, and fantasy while playing, as well as diminished ability to concentrate, focus, and learn. Many modifications result in a child with severely damaged self-confidence, self-worth, and poor performance. A child more or less deprived of self-worth cannot enjoy, give, or receive. A child deprived of emotions turns cold, rational, asocial, socially stiff, uncomfortable, and in the extreme case...intentionally "evil". When a child denies his or her own sex, it becomes invisible, uninteresting, and vague or becomes like the opposite sex in behavior and appearance. The general holistic solution to the vast diversity of symptoms in children with low quality of life is to improve the situation for the child and give the child the holding and support he or she needs. It is very important to realize that a negative belief often has survival value to the child as it helps the child to avoid taking responsibility for problems, which really belong to the parents or other adults. Children have a fine capability for spontaneous healing, and seem to enter this process more easily than adults, given sufficient holding. The symptoms of children with poor thriving ability are often difficult to understand, as they are caused by a complex combination of self-modification in five existential dimensions. This often leads to complex medical diagnosis, giving the idea that the child is sick and without therapeutic reach, while sufficient holding could solve the problem. If holding and support of the child is not enough, the situation must be carefully analyzed to find other possible causes of poor quality of life, health, and functional ability. Education of the parent in holding is often mandatory. Most children with bad thriving ability can thus be helped by simple means. PMID- 14755111 TI - The life mission theory V. Theory of the anti-self (the shadow) or the evil side of man. AB - According to the life mission theory, the essence of man is his purpose of life, which comes into existence at conception. This first purpose is always positive and in support of life. This is not in accordance with the everyday experience that man also engages in evil enterprises born out of destructive intentions. This paper presents a theory about the evil side of man, called the "anti-self" (the shadow), because it mirrors the self and its purpose of life. The core of the anti-self is an evil and destructive intention opposite to the intention behind the life mission. The evil side of man arises when, as the life mission theory proclaims, man is denying his good, basic intention to avoid existential pain. The present theory of the anti-self claims that all the negative decisions accumulated throughout the personal history, sum up to a negative or dark anti self, as complex, multifaceted, and complete as the self. All the negative decisions taken through personal history build this solid, negative, existential structure. The anti-self, or shadow as Carl Gustav Jung used to call it, is a precise reflection of man's basically good and constructive nature. When mapped, it seems that for most or even for all the many fine talents of man, there is a corresponding evil intention and talent in the person's anti-self. As man is as evil as he is good, he can only realize his good nature and constructive talents by making ethical choices. Ethics therefore seem to be of major importance to every patient or person engaged in the noble project of personal growth. Understanding the nature and structure of the evil side of man seems mandatory to every physician or therapist offering existential therapy to his patient. The theory of anti-self makes it possible to treat patients with destructive behavioral patterns, who want to be good deep in their heart, by helping them let go of their evil intentions. The anti-self also seems to explain the enigma of why human beings often commit suicide. Integrating the shadow often leads to dramatic, subjective experiences of ubiquitous light in an "unpersonal" form, of enlightenment, or of meeting light and consciousness in a personal, universal form, known as G-d. PMID- 14755112 TI - Pesticide residues in canned foods, fruits, and vegetables: the application of Supercritical Fluid Extraction and chromatographic techniques in the analysis. AB - Multiple pesticide residues have been observed in some samples of canned foods, frozen vegetables, and fruit jam, which put the health of the consumers at risk of adverse effects. It is quite apparent that such a state of affairs calls for the need of more accurate, cost-effective, and rapid analytical techniques capable of detecting the minimum concentrations of the multiple pesticide residues. The aims of this paper were first, to determine the effectiveness of the use of Supercritical Fluid Extraction (SFE) and Supercritical Fluid Chromatography (SFC) techniques in the analysis of the levels of pesticide residues in canned foods, vegetables, and fruits; and second, to contribute to the promotion of consumer safety by excluding pesticide residue contamination from markets. Fifteen different types of imported canned and frozen fruits and vegetables samples obtained from the Houston local food markets were investigated. The major types of pesticides tested were pyrethroids, herbicides, fungicides, and carbamates. By using these techniques, the overall data showed 60.82% of the food samples had no detection of any pesticide residues under this investigation. On the other hand, 39.15% different food samples were contaminated by four different pyrethroid residues +/- RSD% ranging from 0.03 +/- 0.005 to 0.05 +/- 0.03 ppm, of which most of the pyrethroid residues were detected in frozen vegetables and strawberry jam. Herbicide residues in test samples ranged from 0.03 +/- 0.005 to 0.8 +/- 0.01 ppm. Five different fungicides, ranging from 0.05 +/- 0.02 to 0.8 +/- 0.1 ppm, were found in five different frozen vegetable samples. Carbamate residues were not detected in 60% of investigated food samples. It was concluded that SFE and SFC techniques were accurate, reliable, less time consuming, and cost effective in the analysis of imported canned foods, fruits, and vegetables and are recommended for the monitoring of pesticide contaminations. PMID- 14755113 TI - Syndecan-1 as a regulator of chemokine function. PMID- 14755114 TI - Differences between lovastatin and simvastatin hydrolysis in healthy male and female volunteers:gut hydrolysis of lovastatin is twice that of simvastatin. AB - The aim of this pharmacokinetic evaluation was to show the effect of the extra methyl group in simvastatin on esterase hydrolysis between lovastatin and simvastatin in male and female volunteers. This study was based on the plasma concentration-time curves and the pharmacokinetics of lovastatin and simvastatin with its respective active metabolite statin-beta-hydroxy acid obtained from two different bioequivalence studies, each with 18 females and 18 males. Results were: The group of female volunteers showed a higher yield of the active metabolite beta-hydroxy acid than the group of males (p < 0.002) for both lovastatin and simvastatin. This difference was not related to the body weight of both groups. In the male/female groups, subject-dependent yield of active metabolite beta-hydroxy acid was demonstrated, which was independent of the formulation. The variation in plasma/liver hydrolysis resulted in a fan-shaped distribution of data points when the AUCt lovastatin was plotted vs. that of the beta-hydroxy acid metabolite. In the fan of data points, subgroups could be distinguished, each showing a different regression line and with a different Y intercept (AUCtbeta-hydroxy acid). Lovastatin hydrolysis was higher than simvastatin hydrolysis. It was possible to discriminate between hydrolysis of both lovastatin and simvastatin by plasma/liver or tissue esterase activity. The three subgroups of subjects (males/females) showing different but high yield of statin beta-hydroxy acid can be explained by variable hydrolysis of plasma and hepatic microsomal and cytosolic carboxyesterase activity. This study showed clearly that despite the subject-dependent hydrolysis of lovastatin/simvastatin to the active metabolite, males tend to hydrolyse less than females. The extra methyl group in simvastatin results in less hydrolysis due to steric hindrance. PMID- 14755115 TI - Lack of male-female differences in disposition and esterase hydrolysis of ramipril to ramiprilat in healthy volunteers after a single oral dose. AB - The objective of this study was to identify differences in disposition and esterase hydrolysis of ramipril between male and female volunteers. Plasma concentration and area under the concentration-time curve until the last measured concentration (AUCt) data of ramipril and its active metabolite ramiprilat ( diacid) were obtained from a randomised, cross-over bioequivalence study in 36 subjects (18 females and 18 males). Participants received a single 5-mg oral dose of two different formulations of ramipril (Formulation I and II). Plasma ramipril and ramiprilat concentrations were determined according to validated methods involving liquid chromatography-mass spectrometry. A total number of 2 x 34 available plasma concentration-time curves of both the parent drug and the metabolite could be analysed, and variations (50-100% coefficient of variation [CV]) in plasma concentrations of both parent drug and metabolite were found. With both the formulations, the mean plasma concentrations-time curves of males and females were identical. The groups of female and male volunteers showed similar yields (AUCt = microg x h/L) of the metabolite ramiprilat (p = 0.37); however, females showed a higher AUCt/kg than males (p = 0.046). This difference was solely attributed to the difference in body weight between males and females (p = 0.00049). In both male and female groups, a subject-dependent yield of active metabolite ramiprilat was demonstrated, which was independent of the formulation. There is a large variation in the ramiprilat t1/2beta (50-60% CV). There is a group of subjects who showed a t1/2beta of approximately 80 h (15% CV), and two apparent groups with a longer t1/2beta for each formulation (124 h, 22.5% CV; 166 h, 21.6% CV, respectively, p = 0.0013). This variation in the terminal half-life of ramiprilat is not sex related. In all three groups of half lives, the corresponding Cmax values (mean +/- SD) of ramiprilat in males and females were identical. Thus, with identical Cmax and half-lives, the difference found in the AUCt/kg of ramiprilat must be due to the difference in dose, as the consequence of the difference in body weight, following a standard dose of 5 mg in both males and females. This study showed clearly that despite subject dependent hydrolysis of ramipril to the active metabolite ramiprilat, the variability in the rate of hydrolysis between males and females is similar. With a fixed dose (5 mg), females received a higher dose/kg than males and consequently showed a higher AUCt/kg of the active metabolite ramiprilat. PMID- 14755117 TI - A role for exocytosis in the spatial regulation of actin organization. AB - The efficient organization of the actin cytoskeleton is important for many cellular functions. However, how the local actin organization is regulated in a cell is not well understood. By using yeast mutants defective in actin organization and secretion, we demonstrated that exocytosis plays a role in the spatial regulation of actin organization. Our findings suggest that the actin cytoskeleton, exocytosis, and perhaps endocytosis, may depend on each other for efficiency and reinforce each other. PMID- 14755118 TI - Early discharge after delivery. A study of safety and risk factors. AB - The increased frequency of early discharge of newborns has led to questions of its safety. Most studies have looked at mortality and rehospitalization, not all missed diagnoses. The purpose of this study was to determine diagnoses in newborn infants that would have been missed if the infant had been discharged in <24 h. The design was a cohort study at Rabin Medical Center-Beilinson Campus (average monthly deliveries 1996 [250], 1997 [500]), a university-affiliated community hospital with all in-born term (> or = 37 weeks) infants born September through November 1996 and June 1997. The main outcome measures were medical diagnoses (except trivial physical descriptions) noted at discharge (generally at > or =48 h) exam, not noted on admission exam (<24 h). The results showed that 54 infants (5.1%) had diagnoses that were not detected before the infant was 24 h of age. The leading diagnosis was hyperbilirubinemia. Other potentially missed diagnoses included congenital heart disease (n = 10), morbidity of birth trauma (n = 9), metabolic disturbances (n = 2), hip dislocation (n = 1), suspected sepsis (n = 2), excessive weight loss (n = 2), polycythemia (n = 2), inguinal hernia (n = 1), and abducens paresis (n = 1). It is concluded that diagnoses can be missed by discharging infants in 24 h or less. These diagnoses have the potential for adverse sequela. Even if early discharge is felt to be cost effective, parents should be counseled that it is not risk free. Better mechanisms should be put in place for assuring the safety of such infants. PMID- 14755119 TI - Squeezing an egg into a worm: C. elegans embryonic morphogenesis. AB - We review key morphogenetic events that occur during Caenorhabditis elegans (www.wormbase.org/) embryogenesis. Morphogenesis transforms tissues from one shape into another through cell migrations and shape changes, often utilizing highly conserved actin-based contractile systems. Three major morphogenetic events occur during C. elegans embryogenesis: (1) dorsal intercalation, during which two rows of dorsal epidermal cells intercalate to form a single row; (2) ventral enclosure, where the dorsally located sheet of epidermal cells stretches to the ventral midline, encasing the embryo within a single epithelial sheet; and (3) elongation, during which actin-mediated contractions within the epithelial sheet lengthens the embryo. Here, we describe the known molecular players involved in each of these processes. PMID- 14755120 TI - Child health and human development domain after one year. PMID- 14755121 TI - Holistic medicine IV: principles of existential holistic group therapy and the holistic process of healing in a group setting. AB - In existential holistic group therapy, the whole person heals in accordance with the holistic process theory and the life mission theory. Existential group psychotherapy addresses the emotional aspect of the human mind related to death, freedom, isolation, and meaninglessness, while existential holistic group therapy addresses the state of the person"s wholeness. This includes the body, the person's philosophy of life, and often also love, purpose of life, and the spiritual dimension, to the same extent as it addresses the emotional psyche and sexuality, and it is thus much broader than traditional psychotherapy. Where existential psychotherapy is rather depressing concerning the fundamental human condition, existential holistic therapy conceives life to be basically good. The fundamentals in existential holistic therapy are that everybody has the potential for healing themselves to become loving, joyful, sexually attractive, strong, and gifted, which is a message that most patients welcome. While the patient is suffering and fighting to get through life, the most important job for the holistic therapist is to keep a positive perspective of life. In accordance with these fundamentals, many participants in holistic group therapy will have positive emotional experiences, often of an unknown intensity, and these experiences appear to transform their lives within only a few days or weeks of therapy. An important idea of the course is Bohm's concept of "holo-movement" in the group, resulting from intense coherence between the group members. When the group comes together, the individual will be linked to the totality and the great movement forward towards love, consciousness, and happiness will happen collectively--if it happens at all. This gives the individual the feeling that everything that happens is right, important, and valuable for all the participants at the same time. Native Americans and other premodern people refer to this experience as "the spiritual design". This design is actually an underlying regulation that appears when people, through their feelings and engagement for each other, tie the group together and engage their complex emotional intelligence. Practically, this means that all participants are sunk in the same information matrix, so that everybody learns from each other. Everything that happens in the perception of each trainee has immediate and developing relevance for him. Spontaneous healing happens far more effectively in a group setting, where all the participants stand together and support each other, than it does in the clinic, where the therapist is alone with the patient. A 5-day course in personal development can be compatible to a half year of holistic individual therapy. PMID- 14755122 TI - Factors affecting the formation of a skin reservoir for topically applied solutes. AB - The reservoir function of the skin is an important determinant of the duration of action of a topical solute. The reservoir can exist in the stratum corneum, in the viable avascular tissue (viable epidermis and supracapillary dermis) and in the dermis. A steroid reservoir in the stratum corneum has been demonstrated by the reactivation of a vasoconstrictor effect by occlusion or application of a placebo cream to the skin some time after the original topical application of steroid. Other solutes have also been reported to show a reservoir effect in the skin after topical application. A simple compartmental model is used to understand why reactivation of vasoconstriction some time after a topical steroid application shows dependency on time, topical solute concentration and the product used to cause reactivation. The model is also used to show which solutes are likely to show a reservoir effect and could be potentially affected by desquamation, especially when the turnover of the skin is abnormally rapid. A similar form of the model can be used to understand the promotion of reservoir function in the viable tissue and in the dermis in terms of effective removal by blood perfusing the tissues. PMID- 14755123 TI - Lateral spreading of topically applied UV filter substances investigated by tape stripping. AB - The lateral spreading of topically applied substances is a competitive process to the penetration into the stratum corneum (SC). The penetration of topically applied UV filter substances into the human SC and the lateral spreading were investigated in vivo. Tape stripping in combination with spectroscopic measurements was used to study both processes of two UV filter substances. The concentration of both UV filters was determined inside and outside the application area by varying the application and tape stripping protocol. A spreading of the topically applied substances from the treated to the untreated areas was observed, which caused a concentration gradient. This lateral spreading depends on the time between application and tape stripping and the size of the treated skin area. Significant amounts of topically applied substances were found adjoining the application area, due to the lateral spreading which takes place on the skin surface. In general, the lateral spreading must be considered to be a competitive process when studying penetration processes of topically applied substances. It has to be considered during drug treatment of small limited skin areas and for the interpretation of recovery rates obtained in penetration studies. PMID- 14755124 TI - Analysis of epidermal lipids of the healthy human skin: factors affecting the design of a control population. AB - The intervariability of studies on the lipids of human epidermis and stratum corneum is high because of the different origin of the skin samples and the variety of extraction methods used. In the present work, a high-performance thin layer chromatographic technique has been used to study the parameters age, sex, and anatomical site for their effects on the lipid profiles recovered from healthy epidermal skin biopsy specimens. It was found that sex-related differences were seen at the level of the total ceramide concentration. Observed decreases in lipid concentration, due to ageing, depended on the anatomical site. Therefore, these variables should be controlled in a reproducible and standardized way in order to be able to study the direct relationship between skin condition and barrier lipid composition. Only when this relation is established, results of topical treatment can be scientifically evaluated. PMID- 14755125 TI - Toxicity and antimicrobial activity of a hydrocolloid dressing containing silver particles in an ex vivo model of cutaneous infection. AB - In the present study we examined the effects of two hydrocolloid wound dressings (conventional silver-free Comfeel, silver-incorporating Contreet-H) on uninfected and Candida albicans- or methicillin-resistant Staphylococcus aureus-infected reconstituted human epithelium (RHE). The morphological alterations of the keratinocytes caused by infection and by treatment were analysed with light and electron microscopy. As a measure of epithelial cell damage the release of lactate dehydrogenase from epithelial cells into the surrounding medium was monitored. Application of Contreet-H or Comfeel to uninfected RHE induced no major morphological effects on epithelial cells. Both wound dressings reduced the growth of micro-organisms. Specific alterations of the infected epithelium (vacuoles, spongiosis, oedema, detachment of keratinocytes) and invasion of the epithelium were significantly reduced only by treatment with Contreet-H. At the ultrastructural level release of silver by Contreet-H and superior antimicrobial efficacy could be verified. In summary, treatment with both wound dressings reduced the number of pathogens, with the silver-based wound dressing providing a more effective antimicrobial activity. This resulted in a strong decrease of pathogen-specific alterations of the infected epithelium. We present evidence that delivering silver to infected keratinocytes in a moist healing environment improves the benefit/risk ratio as compared to silver-free wound dressings. PMID- 14755126 TI - Detection of monohydroxyeicosatetraenoic acids and F2-isoprostanes in microdialysis samples of human UV-irradiated skin by gas chromatography-mass spectrometry. AB - UV irradiation of the human skin leads to induction of oxidative stress and inflammation mediated by reactive oxygen radicals, lipid peroxidation, liberation of arachidonic acid from membrane phospholipids and formation of prostaglandins and leucotrienes. We investigated "lipid mediators", such as F(2)-isoprostanes (8 iso-PGF(2alpha), 9alpha,11alpha-PGF(2alpha)) and monohydroxyeicosatetraenoic acids (HETEs) in the dermal interstitial fluid obtained by a cutaneous microdialysis technique. Defined areas on the volar forearm of 10 healthy volunteers were exposed to UVB irradiation (20-60 mJ/cm(2)). Microdialysis membranes were cutaneously inserted beneath the irradiated area. The probes were perfused with isotonic saline solution, and microdialysate samples were collected at 20-min intervals up to 4-5 h. Oxidized arachidonic acid derivatives (2-, 3-, 5 , 8-12- and 15-HETEs, 8-iso-PGF(2alpha) and 9alpha,11alpha-PGF(2alpha)) could be detected and quantified in microdialysates of normal skin in the picomole (HETEs) and femtomole (isoprostanes) range and after UVB irradiation using sensitive gas chromatography-mass spectrometry/negative ion chemical ionization. UVB irradiation enhanced the levels of 8-iso-PGF(2alpha) after 24 h significantly, whereas the HETE levels were slightly increased within shorter time intervals (3 h after UVB irradiation). Further investigations have to show whether these new findings are relevant to validate therapeutic strategies for topical and systemic UV prevention agents or for monitoring of specific therapeutic strategies in inflammatory skin disorders. PMID- 14755127 TI - Photoprotection by Cichorum endivia extracts: prevention of UVB-induced erythema, pyrimidine dimer formation and IL-6 expression. AB - In the gradual process of evolution, plants have developed natural sun protecting substances that enable continuous survival under direct and intense ultraviolet (UV) radiation. As part of our studies of plant-derived pigments that might constitute an alternative to conventional sunscreens, we have tested the ethanolic extracts of roots, stalks, and inflorescences of populations of wild Cichorum endivia subsp. Divaricatum (Asteraceae) in terms of protection against sunburn, and in prevention of UVB-induced pyrimidine dimer formation and IL-6 mRNA expression in the human keratinocyte cell line, HaCaT. Using ELISA technique for detection of pyrimidine dimers and RT-PCR for detection of IL-6, we found that the ethanolic extract of C. endivia roots absorbs radiation in the UVB spectrum and partially prevents induction of pyrimidine dimers and IL-6 expression. Application of the root extract on the skin prior to UVB irradiation totally prevented erythema. Our findings suggest that C. endivia extracts might possess sun-protective qualities that make them useful as sunscreens. PMID- 14755128 TI - Enhancement of wound healing by shikonin analogue 93/637 in normal and impaired healing. AB - Wound healing is a complicated biological process, which involves interactions of multiple cell types, various growth factors, their mediators and the extracellular matrix proteins. In this study, we evaluated the effects of shikonin analogue 93/637 (SA), derived from the plant Arnebia nobilis, on normal and hydrocortisone-induced impaired healing in full thickness cutaneous punch wounds in rats. SA (0.1%) was applied topically daily as an ointment in polyethylene glycol base on wounds. SA treatment significantly accelerated healing of wounds, as measured by wound contraction compared to controls in hydrocortisone-impaired animals. SA treatment promoted formation of granulation tissue including cell migration and neovascularization, collagenization and reepithelialization. The expression of basic fibroblast growth factor (bFGF) was higher as revealed by immunohistochemistry in treated wounds compared to controls. However, the expression of transforming growth factor-beta(1) was not affected by SA treatment. Since bFGF is known to accelerate wound healing, the increased expression of bFGF by SA may be partly responsible for the enhancement of wound healing. These studies suggest that SA could be further studied for clinical use to enhance wound healing. PMID- 14755129 TI - Hypothalamic-pituitary-adrenocortical system and mood disorders: highlights from mutant mice. AB - In recent years, refined molecular technologies and the generation of genetically engineered mice have allowed to specifically target individual genes involved in the regulation of the hypothalamic-pituitary-adrenocortical (HPA) system. Given the fundamental role of the corticotropin-releasing hormone (CRH) system in anxiety, stress-associated pathologies, and mood disorders, we describe genetic modifications of the genes that encode proteins integral to the CRH/CRH receptor system with particular emphasis on conditional gene-targeting strategies. The profile of results, consistent with current knowledge of CRH function from more traditional assays, indicates that enhancement of the CRH function is associated with an activation of the HPA system, an anxious phenotype, alterations in cognitive performance, reductions in food intake, and disturbances of autonomic functions. In general, blockade of CRH activity produces the opposite effects, namely an anxiety-reduced phenotype. Molecular genetic strategies for conditional inactivation or overexpression of the glucocorticoid receptor contribute to our understanding of the genetics of endocrine activity and behavior, the most complex form of biological organization. In addition, we introduce mice with a genetic manipulation in the function of the blood-brain barrier as an animal model for the study of neuroendocrine regulation and, in particular, of HPA system activity. By use of mice deficient for abcb1- (also called multidrug resistance gene 1, mdr1-) type P glycoproteins, it was shown most recently that abcb1-type P glycoproteins control the access of endogenous glucocorticoids into the central nervous system. Thus, the ABCB1-type P glycoprotein function exerts a profound influence on activity and regulation of the HPA system under both basal conditions and during stress. Taken together, these genetically engineered mice are valuable tools for increasing our understanding of HPA system dysregulation in anxiety and stress-related pathologies, including human affective disorders. The identification and detailed characterization of these molecular pathways will ultimately lead to the development of novel neuropharmacological intervention strategies. PMID- 14755130 TI - Agonist-specific coupling of growth hormone secretagogue receptor type 1a to different intracellular signaling systems. Role of adenosine. AB - The growth hormone secretagogue receptor subtype 1a (GHSR-1a) is involved in biological actions of ghrelin by triggering intracellular second messengers coupled to heterotrimeric G-protein complex involving Galpha(q/11). Adenosine is a partial agonist of the GHSR-1a, binding to a binding pocket distinct from the one described for ghrelin. This suggests a variety of functions for the poorly understood GHSR1a receptor. In this work, a sequential analysis of the pathways involved in the regulation of GHSR-1a signaling was undertaken to characterize the intracellular calcium mobilization that is observed following adenosine binding. The results showed that adenosine induced, in a dose-dependent manner, a calcium mobilization from IP(3)-sensitive intracellular stores since the IP(3) receptor blocker 2-APB was able to suppress the calcium response. However, adenosine did not show any effect in the formation of inositol phosphates. The calcium-mobilizing activity was blocked after preincubation of cells with CTX, the inhibitor of adenylate cyclase MDL-12,330A and the protein kinase A blocker H 89. Furthermore, the administration of adenosine stimulated cAMP production. Based on the experimental data, a signaling pathway is proposed involving adenylate cyclase and protein kinase A, which causes phosphorylation of the IP(3) receptor, with a cross-talk between the signaling pathways activated by ghrelin and adenosine. The data described in this report suggest that GHSR-1a is able to activate different intracellular second-messenger systems depending on the agonist that activates it. The regulation of the ghrelin-activated earliest signaling pathways by adenosine may have unexpected implications in the GHSR-1a actions. PMID- 14755131 TI - Arginine increases growth hormone gene expression in rat pituitary and GH3 cells. AB - The effect of arginine (Arg) and Ornitargin (OT) [a compound containing the aminoacids Arg, citrulline (Cit) and ornithine (Orn)] administration upon growth hormone (GH) gene expression was studied both in vivo and in vitro (hemipituitaries and GH3 cells) by Northern blot analysis. For in vivo studies, adult male Wistar rats were anesthetized, subjected to i.v. infusion of 200 microl of 150 mM NaCl (control group), Arg (15 or 150 mg) or OT (15 mg of Arg, 1 mg of Cit and 4 mg of Orn) at a rate of 20 microl/min, and killed 50 min thereafter. For the in vitro studies, hemipituitaries or GH3 cells were incubated in 1 ml of appropriate medium containing Arg (15 or 150 mg) or OT (15 mg of Arg, 1 mg of Cit and 4 mg of Orn) for 60 min. The pituitaries of the in vivo and in vitro studies and GH3 cells were subsequently processed for RNA extraction. Total RNA was subjected to electrophoresis in agarose (1%)/formaldehyde gel, transferred to a nylon membrane and subjected to hybridization with a rat GH (32)P-cDNA, and (32)P-18S rRNA probe to correct for the variability in RNA loading. After autoradiography of the membrane, the abundance of GH mRNA and 18S rRNA bands was quantified by densitometry. The in vivo study demonstrated that Arg and OT infusion induced a 2.3-fold increase in GH mRNA expression, which could result from the Arg-mediated inhibition of somatostatin release. In addition, in vitro Arg, but not OT, induced GH gene expression in hemipituitaries and GH3 cells, indicating that the aminoacid can act per se at the pituitary somatotrope level. In conclusion, our data show for the first time that arginine stimulates GH gene expression in parallel to its recognized GH-releasing activity. PMID- 14755132 TI - Modulation of neuroendocrine response and non-verbal behavior during psychosocial stress in healthy volunteers by the glutamate release-inhibiting drug lamotrigine. AB - The present work was aimed at verifying the following hypotheses: (a) lamotrigine, a drug used to treat mood disorders, affects regulation of stress hormone release in humans, and (b) non-verbal behavior during mental stress situations (public speech) is related to hormonal responses. To achieve these aims, we performed a controlled, double-blind study investigating hormonal responses and non-verbal behavior during public speech in healthy subjects with placebo or lamotrigine (300 mg per os) pretreatment. The stress procedure was performed in 19 young healthy males 5 h following drug or placebo administration. Data were obtained from cardiovascular monitoring, blood and saliva samples, as well as the video-recorded speech. Pre-stress hormone levels were not affected by lamotrigine treatment. Lamotrigine significantly inhibited diastolic blood pressure, growth hormone and cortisol increases during psychosocial stress. In contrast, it potentiated plasma renin activity and aldosterone responses. Non verbal behavior analysis revealed a correlation between catecholamines and submissive or flight behavior in controls, while between catecholamines and displacement behavior following lamotrigine administration. In conclusion, effects of lamotrigine on hormone release might be of value for its mood stabilizing action used in the treatment of bipolar disorder. The data are in support of a stimulatory role of glutamate in the control of cortisol and growth hormone release during psychosocial stress in humans; however, further studies using more selective drugs are needed to prove this suggestion. The effects on plasma renin activity and aldosterone release observed seem to be related to other actions of lamotrigine. PMID- 14755133 TI - Involvement of endogeneous glutamate in the stimulatory effect of norepinephrine and serotonin on the hypothalamo-pituitary-adrenocortical axis. AB - The effects of ionotropic glutamate receptor antagonists on the pituitary adrenal responses following injections of norepinephrine (NE) and serotonin (5-HT) receptor agonists into the hypothalamic paraventricular nucleus (PVN) or electrical stimulation of central NE and 5-HT pathways were studied in anesthetized male rats. PVN injections of an alpha(1)-adrenergic receptor agonist or a serotonergic 5-HT(1A) receptor agonist markedly increased both adrenocorticotropin (ACTH) and corticosterone (CS) serum levels. These responses were significantly inhibited by separate pre-injection of the selective non-NMDA and NMDA glutamate receptor subtype antagonists into the PVN in a dose-dependent manner. Electrical stimulation of either the ventral noradrenergic bundle or the dorsal raphe nucleus markedly increased serum ACTH and CS. These responses were also significantly attenuated by pre-injection of the above glutamate ionotropic receptor antagonists in a dose-dependent manner. These results suggest that glutamatergic interneurons in the PVN, acting via non-NMDA and NMDA receptors, may act as an excitatory mechanism in the NE and 5-HT control of hypothalamic ACTH secretagogues. PMID- 14755134 TI - Mirtazapine decreases stimulatory effects of reboxetine on cortisol, adrenocorticotropin and prolactin secretion in healthy male subjects. AB - Reboxetine is a selective noradrenaline reuptake inhibitor, whereas mirtazapine acts as an antagonist at noradrenergic alpha(2), serotonin (5-HT(2)), 5-HT(3) and histamine H(1) receptors. In a former study we could demonstrate an inhibitory impact of mirtazapine on cortisol secretion. In the present investigation, the influence of combined administration of 15 mg mirtazapine and 4 mg reboxetine on the cortisol (COR), adrenocorticotropin (ACTH), growth hormone (GH), and prolactin (PRL) secretion was examined in 12 healthy male subjects, compared to reboxetine alone (4 mg). In a randomized order, the subjects received reboxetine (4 mg) alone or the combination of reboxetine (4 mg) and mirtazapine (15 mg) at 8:00 a.m. on two different days. After insertion of an intravenous catheter, blood samples were drawn 1 h prior to the administration of single reboxetine or the combination (reboxetine and mirtazapine), at time of administration, and during the time of 5 h thereafter in periods of 30 min. Serum concentrations of COR, GH, and PRL as well as plasma levels of ACTH were determined in each blood sample by means of double antibody RIA, fluoroimmunoassay and chemiluminescence immunometric assay methods. The area under the curve (AUC) was used as parameter for the COR, ACTH, GH, and PRL response. For statistical evaluation, the Wilcoxon signed-ranks test was performed. There was a pronounced stimulation of COR, ACTH, GH, and PRL concentrations after single administration of reboxetine. When reboxetine was given in combination with mirtazapine, a significant reduction of the COR, ACTH, and PRL stimulation was observed whereas GH secretion patterns remained unchanged, compared to single administration of reboxetine. Apparently, the stimulatory effects of reboxetine on pituitary hormone secretion via noradrenergic mechanisms are counteracted in part by the alpha(2)-blocking properties of mirtazapine and its inhibitory influence on cortisol secretion. PMID- 14755135 TI - Inhibition of Ras-GTPase improves diabetes-induced abnormal vascular reactivity in the rat perfused mesenteric vascular bed. AB - OBJECTIVE: The signalling mechanisms involved in regulating altered vascular reactivity in diabetes are not fully understood. The aim of this study was to investigate the role of Ras-GTPase in the development of abnormal vascular reactivity in diabetes. MATERIALS AND METHODS: We investigated the ability of chronic administration of FPTIII (1.5 mg/kg), an inhibitor of Ras-GTPase, to modulate the altered vasoreactivity of the rat perfused mesenteric bed to common vasoconstrictors and vasodilators in streptozotocin (STZ)-induced diabetes. RESULTS: The vasoconstrictor responses induced by norepinephrine (NE) and endothelin-1 (ET-1) were significantly increased whereas vasodilator responses to carbachol, histamine and isoprenaline were significantly reduced in the perfused mesenteric bed of the STZ-diabetic rats. Inhibition of Ras-GTPase by chronic administration of FPTIII produced a significant normalization of the altered agonist-induced vasoconstrictor and vasodilator responses without affecting blood glucose levels. Inhibition of Ras-GTPase did not affect the agonist-induced vasoconstrictor and vasodilator responses in the control animals. CONCLUSION: These data suggest that signal transduction pathways activated by Ras-GTPase are involved in the development of diabetic vascular dysfunction. Potential strategies aimed at modifying actions of signal transduction pathways involving Ras-GTPase may therefore prove to be beneficial in treatment of vascular complications in diabetes. PMID- 14755136 TI - Differential distribution of risk factors and outcome of acute coronary syndrome in Kuwait: three years' experience. AB - OBJECTIVE: To compare the distribution of risk factors and clinical outcome of acute coronary syndrome (ACS) between Kuwaiti and other Arab men living in Kuwait. SUBJECTS AND METHODS: The data for this study was collected from the computerized database at the Mubarak Al-Kabeer Hospital, Kuwait and the 1997-2000 census data for the State of Kuwait. 1,329 Arab men (666 Kuwaitis and 663 other Arabs) older than 25 years who were admitted between September 1997 and August 2000 with a diagnosis of ACS were included in the study. RESULTS: The rate of admission for the entire patient population was twofold higher for Kuwaiti (1.68/1,000) than other Arab men (0.72/1,000), (p < 0.001); the mean age of the Kuwaiti men was 56.7 +/- 11.9 years and other Arab men 53.0 +/- 10.5 years (p < 0.001). The prevalence of hypertension, diabetes, smoking and hypercholesterolemia for Kuwaiti men was 35.9, 56.9, 51.7 and 36.2%, respectively; the corresponding prevalence for other Arab men was 28.8, 42.7, 68.2 and 32.0%, the difference in the prevalence except for hypercholesterolemia was significant (p < 0.001). In Kuwaiti men younger than 55 years of age, the prevalence of hypertension, diabetes mellitus, smoking and hypercholesterolemia was 26.6, 49.5, 68.6 and 43.3%, respectively; the corresponding values for other Arab men was 22.3, 36.2, 77.7 and 43.3%; the difference in prevalence except for hypertension was significant (p < 0.001). The in-hospital mortality for the whole study was 6.2% (Kuwaiti) and 2.3% (other Arab men; p < 0.001); while that for men younger than 55 years was 2.7% (Kuwaiti) and 0.8% (other Arab men; p < 0.05). CONCLUSION: The rate of admission for the entire patient population with a diagnosis of ACS was twofold higher for Kuwaiti than other Arab men. Among all patients and also those less than 55 years, the prevalence of diabetes mellitus was consistently higher among Kuwaiti than other Arab men thereby probably leading to the higher admission rate and in-hospital mortality. PMID- 14755137 TI - Primary malignant tumors of the trachea - the Tata Memorial Hospital experience. AB - OBJECTIVE: Primary tumors of the trachea are extremely rare. Treatment methods vary considerably and few studies have sought to provide adequate guidelines. This study reviews the records of patients treated for tracheal cancer at the Tata Memorial Hospital (TMH), Mumbai, India. SUBJECTS AND METHODS: Fifteen patients with primary tracheal malignancies were identified in the TMH database during the period from 1983 to 2000. They were predominantly males (87%) belonging to an older age-group (67% above 40 years). Common presenting symptoms were cough, hoarseness, hemoptysis and indications of airway obstruction. Squamous cell carcinoma was the commonest histologic subtype (40%) followed by adenoid cystic carcinoma (27%). Ten patients received radical treatment. One patient underwent surgery (resection and anastomosis) and received postoperative radiotherapy. Another was explored but was found to be unresectable and was 1 of 2 patients treated with chemotherapy and radiotherapy. Laser resections and radiotherapy were used in 2 patients while 4 patients were managed with radiotherapy alone. One patient was treated elsewhere. The majority of patients (8/9) were treated with locoregional fields and doses ranging from 40 to 60 Gy (median 50 Gy). Two patients also received intraluminal brachytherapy, 1 as part of initial treatment and another for recurrence. RESULTS: Only 5 patients treated at TMH (5/9) achieved local control of their disease. Follow-up times ranged from 1 month to 134 months, median of 38 months. Distant metastases were identified in 4 patients (bone n = 1 and lung n = 3). Median survival was 38 months. Overall survival at 5 years was 37% by Kaplan-Meier method, but this figure should be treated with caution since only 6 patients had a follow-up of more than 2 years. CONCLUSION: Tracheal cancer is a rare malignancy. Radiation therapy is a reasonably effective modality for unresectable disease. PMID- 14755138 TI - Pretherapy Gallium-67 scanning in paediatric patients with Hodgkin's disease. AB - OBJECTIVE: To investigate the correlation between gallium-67 (67Ga) uptake and histological subtypes of Hodgkin's disease (HD) in paediatric patients. SUBJECTS AND METHODS: Fifty-eight patients (45 males and 13 females aged 9.2 +/- 4 years, range 1.5-17 years) with histologically diagnosed HD underwent pretherapy 67Ga scanning on days 2, 5, 12 or 14 days after intravenous administration of 25-50 MBq (0.7-1.4 mCi) of 67Ga citrate. The scans were evaluated both visually and quantitatively using the activity of 67Ga in the liver as a reference. Clinical outcome of 11 patients with high diffuse 67Ga skeletal uptake was compared with that of 17 patients showing normal distribution of 67Ga in the skeleton. RESULTS: Of the 58 patients, the 67Ga scans were positive in 47 patients with 117 lesions. Visual analysis did not differentiate between the histological variants of HD. However, quantitative analysis of lesion-to-liver ratios showed significantly higher values of 67Ga uptake in the mixed cellularity type than in the nodular sclerosis type (t = -3.7, p < 0.001). Patients with high skeletal uptake had a higher relapse rate (6/11) than those with normal skeletal uptake (3/17). CONCLUSION: The findings show that quantitative analysis of 67Ga uptake can differentiate between the two main subtypes of HD (mixed cellularity and nodular sclerosis). Further diffuse skeletal uptake of 67Ga indicates a higher relapse rate. PMID- 14755139 TI - Clinical patterns of diffuse parenchymal lung disease in Kuwait: a prospective study. AB - OBJECTIVE: To report our experience of the clinicopathological patterns of diffuse parenchymal lung disease (DPLD). SUBJECTS AND METHODS: Over a 4-year period, 75 patients (41 males, 34 females), aged 13-76 years, who were referred to Mubarak Al-Kabeer and the Chest Diseases Hospitals, Kuwait with a diagnosis of diffuse lung disease, were included in the study. After a comprehensive history and physical examination, further investigations were done, including hematological and immunological profiles, sputum and bronchoalveolar lavage fluid examination, chest radiograph, high resolution computed tomography (HRCT), pulmonary function test and lung biopsy. RESULTS: Of the 75 patients 60 (80%) were over 40 years of age. The duration of symptoms in 34 patients (45%) was less than 6 months and longer than 1 year in 28 (37.7%) patients. Twenty-five of the patients were cigarette smokers. The mean forced lung capacity (FVC), total lung capacity and diffusing capacity for carbon monoxide were less than 60% of the predicted values in most patients. There was a significant difference in mean FVC value between smokers and nonsmokers (p < 0.05). The HRCT findings were at an advanced stage in 65 patients, with additional honeycombing in 21 of the 65 patients. Idiopathic pulmonary fibrosis was the most common cause of DPLD, occurring in 52 patients, followed by sarcoidosis and collagen vascular diseases. CONCLUSION: DPLD was observed predominantly in middle aged and elderly patients, due probably to increasing industrialization in the country. The role of cigarette smoking as a contributory factor remains unclear. PMID- 14755140 TI - Oxidative stress in patients with chronic renal failure: effects of hemodialysis. AB - OBJECTIVE: To investigate blood oxidative status of patients with chronic renal failure (CRF) and possible effects of hemodialysis on the development of oxidative stress in blood. MATERIALS AND METHODS: The levels of malondialdehyde (MDA) and oxidation resistance (OR) values were measured in blood plasma, erythrocyte hemolysate and erythrocyte membrane fractions of 33 patients with CRF and of 12 healthy controls. Of the 33 patients, 17 subjects were under hemodialysis treatment. RESULTS: MDA levels were found to be increased in all blood fractions of the patients. OR values were unchanged in erythrocyte hemolysates but decreased in plasma and erythrocyte membrane fractions of the CRF patients. Moreover, erythrocyte MDA levels were determined to be higher in hemodialyzed patients compared with both controls and non-hemodialyzed patients. OR values were lower in all blood fractions of the hemodialyzed patients relative to controls and non-hemodialyzed patients. CONCLUSION: Results suggest that there is a significant oxidative stress (expressed as peroxidation) in blood samples from patients with CRF, which is further exacerbated by hemodialysis. PMID- 14755141 TI - Monoclonal gammopathy among patients with chronic hepatitis C infection. AB - OBJECTIVE: To determine the prevalence of monoclonal gammopathies in Kuwait and its association with chronic hepatitis C virus (HCV) infection. METHODOLOGY: Serum protein electrophoresis and measurement of immunoglobulin levels were carried out prospectively in 100 consecutive patients with chronic HCV infection. RESULTS: Among the 100 patients tested (82 males, 18 females; median age 45 years), 59 had polyclonal band in serum protein electrophoresis while the other 41 had a normal pattern. None of the patients had monoclonal gammopathy. The mean serum immunoglobulin levels were: IgG 18.4 g/l (range 4-39), IgA 2.9 g/l (range 0.09-8) and IgM 1.6 g/l (range 0.14-6.08). CONCLUSION: There was no evidence of monoclonal gammopathies in patients with chronic HCV infection in Kuwait. PMID- 14755142 TI - Assessment of Chlamydia trachomatis prevalence by cell culture and transcription mediated amplification in symptomatic women. AB - OBJECTIVE: The frequency of Chlamydia trachomatis in women with mucopurulent discharge was determined by a cell culture technique and a transcription-mediated amplification (TMA) assay in endocervical swab specimens. SUBJECTS AND METHODS: Endocervical swab specimens were obtained from 116 symptomatic patients with genitourinary complaints or abdominal pain. All of the women were married, with an age range of between 19 and 44 (median 29) years. The cell culture assay was used in all specimens. For 75 specimens the TMA assay was also performed. RESULTS: Positive cell culture test results were obtained in 6 (5.2%) patients. Among 75 specimens, 2 were positive by both TMA and culture assays, while 1 specimen was positive only by the culture assay. Of those positive for C. trachomatis, 5 were in the 19- to 25-year age group, and 1 was in the >25-year age group. All of the patients with positive results were of low socioeconomic status. CONCLUSIONS: This study revealed a relatively low rate of C. trachomatis infections in symptomatic married women in Turkey. A commercial TMA assay failed to identify all positive patients, in contrast to a 'gold standard' culture assay used in patients having such infections. PMID- 14755143 TI - Role of fine needle aspiration cytology in the diagnosis of swellings in the salivary gland regions: a study of 712 cases. AB - INTRODUCTION: A mass in the salivary gland region often presents a diagnostic challenge with regard to its site of origin (salivary versus nonsalivary), benign or malignant nature, and tissue-specific diagnosis. The present study describes the utility of fine-needle aspiration (FNA) cytology in the diagnosis of these lesions. SUBJECTS AND METHODS: Over a 6-year period (January 1994 to December 1999), 712 patients aged between 6 months and 91 years (median, 37 years) were subjected to FNA of swellings in their salivary gland regions. Male:female ratio was 1.28:1. The swellings were mostly located in the parotid (323 cases), submandibular (343 cases), and upper cervical region (27 cases). Swellings of oral (5 cases) and sublingual (2 cases) sites were rare. The lesions diagnosed by FNA cytology were compared among the major salivary glands. Cytologic diagnoses were correlated with histology in 45 cases. RESULTS: Benign nonneoplastic lesions were the most common (73%), followed by neoplasms (20%), and those with atypical cytology (1%). Cytologic material was inadequate in 6% cases. Parotid gland region was involved more frequently by neoplasms (27.1%) than the submandibular gland region (13.7%, p < 0.0001). Inflammatory processes affected the submandibular gland region more commonly (42.0%) than the parotid (32.6%, p = 0.0164). Pleomorphic adenoma was the most common neoplasm (61.5%), followed by Warthin's tumor (12.6%). Malignancies accounted for 10.5% of neoplasms. Frequency of involvement of parotid by Warthin's tumor (16.7%) was significantly higher than that of submandibular gland (2.3%, p = 0.0191). However, the submandibular gland was more commonly affected by malignancy than the parotid gland (p = 0.0003). Sensitivity, specificity, and diagnostic accuracy of FNA cytology for all neoplastic lesions of the salivary gland were 94.6, 75.0, and 91.1%, respectively. The corresponding figures for malignancies were 60.0, 95.0, and 91.1%, respectively. CONCLUSION: FNA cytology is very useful for the diagnosis of salivary gland lesions. However, sampling and interpretation errors may occur. The low specificity for the diagnosis of neoplasms as a whole and the poor sensitivity for malignancies found in our study can be attributed to the relatively small number of benign nonneoplastic and malignancy cases with available histopathologic diagnoses. PMID- 14755144 TI - Perinatally acquired tuberculosis in a neonate. a case report. AB - OBJECTIVES: To report a case of perinatal tuberculosis that appeared on the 21st day of life of an infant born to a mother with latent tuberculosis. CLINICAL PRESENTATION AND INTERVENTION: A preterm male infant was born by spontaneous vertex delivery at 33 weeks gestational age to a 33-year-old primiparous Philippine woman. The infant was well until the 21st day of life when he developed recurrent episodes of cyanosis and bradycardia. A chest radiograph showed infiltrates which were thought to be bacterial in origin. Blood, urine, and cerebrospinal fluid cultures were normal. Tracheal aspirate revealed acid fast bacilli by Ziehl-Neelsen stain, later confirmed to be MYCOBACTERIUM TUBERCULOSIS by culture in Lowenstein-Jensen medium. The mother was later diagnosed as a case of tuberculosis with symptoms, signs and radiologic manifestation of hilar lymphadenopathy with mild pleural effusion and positive tuberculin skin test. Both infant and mother were treated with intravenous isoniazid, intravenous rifampicin, oral pyrazinamide, and intravenous pyridoxine. Both recovered. CONCLUSION: A preterm male infant perinatally acquired tuberculosis, most likely by inhalation of the bacteria during delivery. Both infant and mother responded well to antituberculous treatment. PMID- 14755145 TI - Giant testicular mixed germ cell tumor. a case report. AB - OBJECTIVE: We report a case that we believe to be the largest example of a testicular mixed germ cell tumor with a clearly defined histology pattern. CLINICAL PRESENTATION AND INTERVENTIONS: A 21-year-old patient consulted a urologist concerning a giant testicular mass. At the time of presentation the tumor measured 29 x 20 x 16 cm, with a weight of 4,850 g. Serum alpha-fetoprotein was over 15,000 ng/ml, while beta-human chorionic gonadotropin was normal. Chest X-ray and CT revealed multiple bilateral metastases. Histopathology revealed a mixed germ cell tumor containing 80% of yolk sack tumor, 10% of teratoma and 10% of embryonal carcinoma. Orchiectomy and chemotherapy were successful in the treatment of primary tumor and bilateral lung metastases. CONCLUSION: This tumor grew to extraordinary dimensions because of the patient's failure, due to fear, lack of knowledge and embarrassment, to seek medical help and to health professionals' negligence when performing regular check-ups. PMID- 14755146 TI - No more intravenous procaine for pancreatitis pain? PMID- 14755147 TI - Procaine hydrochloride fails to relieve pain in patients with acute pancreatitis. AB - BACKGROUND: Several analgesics are in use for pain control in patients with acute pancreatitis. Procaine hydrochloride (procaine) has a long tradition and is recommended by the German Society of Gastroenterology and Metabolic Diseases for pain treatment in patients with acute pancreatitis. There is no controlled trial showing that procaine could be effective for pain treatment. METHODS: In an open, randomized, controlled trial, 107 patients (76 male, 31 female; mean age 45 +/- 12 years) were included and randomized either to receive procaine (n = 55) or pentazocine (n = 52) for pain relief. Procaine 2 g/ 24 h was administered by continuous intravenous infusion, pentazocine 30 mg was administered every 6 h as a bolus intravenous injection. Pentazocine was additionally administered on demand whenever required in patients of both treatment groups and its total consumption was recorded. Pain scores were assessed twice daily on a visual analogue scale. RESULTS: Patients receiving procaine were significantly more likely to request additional analgesics compared to patients treated with pentazocine alone, 98 vs. 44%, respectively (p < 0.001). Procaine did not reduce the amount of pentazocine required for pain control. The amount of pentazocine given in both groups was not statistically significantly different. Recorded pain scores were significantly lower (p < 0.001) in patients in the pentazocine group during the first 3 days of analgesic treatment. From day 4 on there was no significant difference in pain scores among the two groups. CONCLUSION: Thus, intravenous procaine treatment is not effective for pain control in patients with acute pancreatitis. PMID- 14755148 TI - Health care resource utilization and costs of NSAID-induced gastrointestinal toxicity. A population-based study in Switzerland. AB - BACKGROUND AND AIMS: The well-recognized gastrointestinal toxicity of NSAIDs ranges from mild symptoms to severe complications requiring medical attention and leading to work loss. The present study evaluates the economic burden derived from health care resource utilization and work loss caused by NSAID-induced gastrointestinal toxicity in the Swiss population. The costs induced by conventional and COX2-selective NSAIDs were calculated separately, and the results were then extrapolated for the United States. METHODS: A computer generated random sampling of the Swiss population (n = 6,118) was administered a questionnaire by phone. Questions investigated NSAID intake, occurrence of adverse effects, consumption of gastroprotective drugs, health care resource utilization and work loss. Country-specific direct and indirect costs associated with NSAID-induced gastrointestinal toxicity were calculated according to current reimbursement costs in Switzerland and the United States. Medication expenses were used to compute separate iatrogenic cost factors for conventional and COX2 selective NSAIDs. The ability of demographic variables and symptoms to predict resource utilization was analyzed by univariate and multivariate analyses. RESULTS: From among the 6,085 responders (99.5% response rate), 294 subjects had gastrointestinal adverse effects associated with NSAID intake. The annual amount spent on NSAID-related adverse effects was CHF 432.2 million for Switzerland and USD 7,425.7 million for the United States. This amount corresponded to CHF 581 and USD 272 per NSAID user, and was primarily due to costs derived from work loss. For Switzerland, 91.1% of total costs were associated with the use of conventional NSAIDs and could be saved by using COX2-NSAIDs instead. While additional 13% of the medication purchase costs are spent for gastrointestinal adverse effects among COX2-selective NSAID users, additional 315% are spent for the same reason among conventional NSAID users. Thus, given similar efficacy and purchase costs in the two drug groups, COX2-selective NSAIDs are 25.1 times more cost-effective than conventional NSAIDs. Language region, place of residence, nationality, marital status, epigastric pain, diarrhea, hematemesis and melena predicted resource utilization. CONCLUSIONS: Costs arising from NSAID-induced gastrointestinal toxicity are considerable and primarily associated with conventional NSAIDs. COX2-selective NSAIDs could decrease this economic burden. PMID- 14755149 TI - The dawning of a new sedative: propofol in gastrointestinal endoscopy. AB - BACKGROUND: Using gentler endoscopes and improved sedation, great strides have been made in enhancing patients' comfort and acceptance of endoscopic procedures. Because morbidity and mortality have been associated with benzodiazepines in endoscopic sedation, safer alternatives were sought. Propofol (2,6 diisopropylphenol), a rapid and short-acting anesthetic, initially used in the 1980's for general anesthesia induction and maintenance, is a promising candidate. METHODS: This review article examines experiences and literature references of propofol's use in endoscopic procedures. Three critical questions are posed: What are the major advantages and potential risks of propofol? When should propofol be used? Who should administer propofol, how should it be administered, and what type of monitoring is required? RESULTS: With considerable inter-patient variability, the propofol dose must be carefully titrated according to the individual patient's response. Factors influencing dosage include age, ASA class, patient's height and procedure duration. Propofol's primary risk is its narrow therapeutic range which necessitates careful patient monitoring. CONCLUSIONS: Propofol's advantages over benzodiazepines and narcotics include a more rapid onset of action, full relief of discomfort and rapid recovery to alertness without residual sedative effects or anterograde amnesia, thereby making this drug a cost-effective and, with proper monitoring, safe choice. PMID- 14755150 TI - Quantitative assessment of gastric antrum atrophy shows restitution to normal histology after Helicobacter pylori eradication. AB - BACKGROUND/AIMS: Grading gastric mucosal atrophy in antrum biopsy specimens remains a controversial subject because of limitations in interobserver agreement. We previously described a reliable, quantitative method for grading atrophy of the corpus mucosa with excellent reproducibility and good correlation with the Sydney scores. The aims of the present study were to evaluate the applicability of this method for the grading of antral atrophy and to study the effect of Helicobacter pylori eradication on the antral mucosa. METHODS: Antrum biopsy specimens were collected from 71 gastroesophageal reflux disease patients at baseline and after 3 and 12 months. After the first endoscopy, all subjects were treated with omeprazole 40 mg daily for 12 months. After randomization, 27 of the 48 H. pylori-positive patients additionally received eradication therapy. In 182 hematoxylin-eosin-stained specimens, which were of sufficient quality, the proportions (volume percentages) of glands (VPGL), stroma (VPS), infiltrate (VPI), and intestinal metaplasia in the glandular zone of the antrum mucosa were measured using a point-counting method. In these specimens, mucosal atrophy was assessed by two experienced gastrointestinal tract pathologists (E.B. and J.L.) according to the updated Sydney classification as either nonatrophic mucosa (n = 47) or as mild (n = 29), moderate (n = 50), or marked (n = 56) atrophy. In addition, a group of 23 cases with difficult-to-classify grades of atrophy were included. RESULTS: The mean VPGL decreased with increasing Sydney grades of atrophy (p < 0.001), while the mean VPS and VPI increased (both p < 0.001). After H. pylori eradication, even the cases with the lowest VPGL regressed to normal levels. CONCLUSIONS: Overall, a low VPGL correlates with increasing grades of antrum mucosal atrophy. The present data indicate that gastric mucosal atrophy is reversible, since almost all cases showed regression of VPGL after H. pylori eradication. The cases with difficult-to-classify grades of atrophy showed significantly lower VPGLs and higher VPIs than the reference cases. PMID- 14755151 TI - Role of fibroblast growth factor-2 in the expression of matrix metalloproteinases and tissue inhibitors of metalloproteinases in human intestinal myofibroblasts. AB - BACKGROUND AND AIMS: The coordinated expression of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) plays a crucial role in tissue remodeling. We investigated the effects of fibroblast growth factor (FGF)-2 on the secretion of MMPs and TIMPs in human intestinal subepithelial myofibroblasts (SEMFs). METHODS: The secretion of MMP-s and TIMPs was determined by ELISA or Western blotting. The mRNA expression of MMPs and TIMPs was assessed by Northern blotting. The activating protein (AP)-1-DNA binding activity was evaluated by electrophoretic gel mobility shift assays (EMSA). RESULTS: Unstimulated intestinal SEMFs constitutively secreted MMP-2 and TIMP-2. FGF-2 stimulated MMP-1, MMP-3 and TIMP-1 secretion, but did not affect MMP-2 or TIMP-2 secretion. FGF-2 induced AP-1-DNA binding activity, and the c-Jun/AP-1 inhibitor curcumin attenuated the FGF-2-induced MMP-1, -3 and TIMP-1 mRNA expression. Mitogen-activated protein (MAP) kinase inhibitors (U0126 and PD098059) also blocked the MMP-1, -3 and TIMP-1 secretion. Furthermore, FGF-2 dose-dependently induced FGF-2 mRNA expression in these cells. CONCLUSIONS: FGF-2 may be one of important regulatory factors for extracellular matrix turnover via a modulation of MMP and TIMP secretion from SEMFs. PMID- 14755152 TI - Treatment of functional dyspepsia with a herbal preparation. A double-blind, randomized, placebo-controlled, multicenter trial. AB - BACKGROUND: We aimed to assess the efficacy and safety of a herbal preparation STW 5-II containing extracts from bitter candy tuft, matricaria flower, peppermint leaves, caraway, licorice root and lemon balm for the treatment of patients with functional dyspepsia. METHODS: 120 patients with functional dyspepsia were randomly assigned to 1 of 4 treatment groups. Each patient received the treatment for three consecutive 4-week treatment blocks. The first two treatment blocks were fixed. For the third treatment period, medication was based upon the investigator's judgement of symptom improvement during the preceding treatment period. In patients without adequate control of symptoms, the treatment was switched, or if symptoms were controlled, the treatment was continued. The primary outcome measure was the improvement of a standardized gastrointestinal symptom score (GIS). FINDINGS: During the first 4 weeks, the GIS significantly decreased in subjects on active treatment compared to the placebo (p < 0.001). During the second 4-week period, symptoms further improved in subjects who continued on active treatment or who switched to the active treatment (p < 0.001), while symptoms deteriorated in subjects who switched to placebo. After 8 weeks 43.3% on active treatment and 3.3% on placebo reported complete relief of symptoms. (p < 0.001 vs. placebo). CONCLUSION: In patients with functional dyspepsia, the herbal preparation tested improved dyspeptic symptoms significantly better than placebo. PMID- 14755153 TI - Probiotic treatment increases salivary counts of lactobacilli: a double-blind, randomized, controlled study. AB - BACKGROUND/AIMS: Lactobacilli are used in the prevention and treatment of several diseases, but they are also known to play a role in the pathogenesis of dental caries. The aim of our study was to evaluate whether the oral administration of lactobacilli could change the salivary counts of these bacteria compared with placebo. Moreover, lactobacilli were administered in liquid and in capsule form to determine the role of direct contact with the oral cavity. METHODS: Thirty five healthy volunteers were randomized into three groups to receive lactobacilli and/or placebo for 45 days: group A (n = 14) received probiotics in capsules and placebo in liquid form; group B (n = 16) took liquid probiotics and placebo in capsules, and group C (n = 5) used placebo in both liquid and capsule form. Streptococcus mutans populations served as control. The salivary counts of lactobacilli and S. mutans were measured semi-quantitatively using the CRT bacteria kit. RESULTS: Compared with placebo, the oral administration of probiotics, both in capsules and in liquid form, significantly increases salivary counts of lactobacilli (p = 0.005 and p = 0.02, respectively). S. mutans populations were not significantly modified. CONCLUSIONS: The increased salivary counts of lactobacilli may indicate the need to closely monitor the dental health of patients undergoing long-term probiotics treatment, even when this treatment is administrated in a form that avoids direct contact with the oral cavity. PMID- 14755154 TI - Cronkhite-Canada syndrome containing colon cancer and serrated adenoma lesions. AB - We describe a case of Cronkhite-Canada syndrome associated with sigmoid colon cancer, and provide a literature review. A 77-year-old man was diagnosed with sigmoid colon cancer after presenting with hypoproteinemia, nail atrophy, loss of scalp hair, hyperpigmentation, and gastrointestinal polyposis. The findings were consistent with Cronkhite-Canada syndrome. The colon polyps were histologically serrated adenomas, whose crypts showed a saw-toothed growth pattern with dysplasia, or tubular adenoma. Cronkhite-Canada syndrome associated with colon cancer has been reported in 31 cases. The availability of histologic material permitted reexamination of 25 of these cases. Serrated adenoma of the polypoid lesions was retrospectively found in 10 (40%) of the 25 cases. By comparison, the incidence of serrated adenomas has been estimated to occur in about 1% of all general polyps. Taken together, it is suggested that Cronkhite-Canada syndrome associated with colorectal cancer frequently has polyps containing serrated adenoma lesions. In the case described here, microsatellite instability and overexpression of the p53 protein were found in the cancer lesion and serrated adenoma lesions, and none of the lesions showed a loss of heterozygosity of various genes or K-RAS mutations. Thus, genetic alterations between the serrated adenoma and the colorectal cancer was correlated in this case. These findings suggested the possibility of a serrated adenoma-carcinoma sequence in this case of Cronkhite-Canada syndrome. PMID- 14755155 TI - Measurement of dipolar cross-correlation in methylene groups in uniformly 13C-, 15N-labeled proteins. AB - A CC(CO)NH TOCSY-based 3D pulse scheme is presented for measuring (1)H-(13)C dipole-dipole cross-correlated relaxation at CH(2) positions in uniformly (13)C-, (15)N-labeled proteins. Simulations based on magnetization evolution under relaxation and scalar coupling interactions show that cross-correlation rates between (1)H-(13)C dipoles in CH(2) groups can be simply obtained from the intensities of (13)C triplets. The normalized cross-correlation relaxation rates are related to cross-correlation order parameters for macromolecules undergoing isotropic motion, which reflect the degrees of spatial restriction of CH(2) groups. The study on human intestinal fatty acid binding protein (131 residues) in the presence of oleic acid demonstrates that side chain dynamics at most CH(2) positions can be characterized for proteins less than 15 kDa in size, with the proposed TOCSY-based approach. PMID- 14755156 TI - GFT NMR experiments for polypeptide backbone and 13Cbeta chemical shift assignment. AB - (4,3)D, (5,3)D and (5,2)D GFT triple resonance NMR experiments are presented for polypeptide backbone and (13)C(beta) resonance assignment of (15)N/(13)C labeled proteins. The joint sampling of m = 2, 3 or 4 indirect chemical shift evolution periods of 4D and 5D NMR experiments yields the measurement of 2(m) - 1 linear combinations of shifts. To obtain sequential assignments, these are matched in corresponding experiments delineating either intra or interresidue correlations. Hence, an increased set of matches is registered when compared to conventional approaches, and the 4D or 5D information allows one to efficiently break chemical shift degeneracy. Moreover, comparison of single-quantum chemical shifts obtained after a least squares fit using either the intra or the interresidue data demonstrates that GFT NMR warrants highly accurate shift measurements. The new features of GFT NMR based resonance assignment strategies promise to be of particular value for establishing automated protocols. PMID- 14755157 TI - A selective intra-HN(CA)CO experiment for the backbone assignment of deuterated proteins. AB - A modified version of the backbone TROSY based 3D HN(CA)CO experiment is presented that provides uniquely intraresidue correlation shift information, by efficiently suppressing the sequential connectivity data. The resulting reduction in the number of peaks decreases overlap and removes ambiguities in situations where in the conventional HN(CA)CO i and i - 1 correlations are overlapped or are comparable in intensity. The sequential residue connection is obtained in combination with a 3D HNCO. MQ- and SQ-implementations of the experiment were tested on a deuterated protein with a correlation time of 28 ns ( approximately 60 kDa). PMID- 14755158 TI - Solution structure and backbone dynamics of the pleckstrin homology domain of the human protein kinase B (PKB/Akt). Interaction with inositol phosphates. AB - The programmed cell death occurs as part of normal mammalian development. The induction of developmental cell death is a highly regulated process and can be suppressed by a variety of extracellular stimuli. Recently, the ability of trophic factors to promote survival have been attributed, at least in part, to the phosphatidylinositide 3'-OH kinase (PI3K)/Protein Kinase B (PKB, also named Akt) cascade. Several targets of the PI3K/PKB signaling pathway have been identified that may underlie the ability of this regulatory cascade to promote cell survival. PKB possesses a N-terminal Pleckstrin Homology (PH) domain that binds specifically and with high affinity to PtIns(3,4,5)P(3) and PtIns(3,4)P(2), the PI3K second messengers. PKB is then recruited to the plasma membrane by virtue of its interaction with 3'-OH phosphatidylinositides and activated. Recent evidence indicates that PKB is active in various types of human cancer; constitutive PKB signaling activation is believed to promote proliferation and increased cell survival, thereby contributing to cancer progression. Thus, it has been shown that induction of PKB activity is augmented by the TCL1/MTCP1 oncoproteins through a physical association requiring the PKB PH domain. Here we present the three-dimensional solution structure of the PH domain of the human protein PKB (isoform beta). PKBbeta-PH is an electrostatically polarized molecule that adopts the same fold and topology as other PH-domains, consisting of a beta sandwich of seven strands capped on one top by an alpha-helix. The opposite face presents three variable loops that appear poorly defined in the NMR structure. Measurements of (15)N spin relaxation times and heteronuclear (15)N[(1)H]NOEs showed that this poor definition is due to intrinsic flexibility, involving complex motions on different time scales. Chemical shift mapping studies correctly defined the binding site of Ins(1,3,4,5)P(4) (the head group of PtIns(3,4,5)P(3)), as was previously proposed from a crystallographic study. More interestingly, these studies allowed us to define a putative alternative low affinity binding site for Ins(1,4,5)P(3). The binding of this sugar to PKBbeta-PH might also involve non-specific association that could explain the stabilization of the protein in solution in the presence of Ins(1,4,5)P(3). PMID- 14755159 TI - Heteronuclear 2D (1H-13C) MAS NMR resolves the electronic structure of coordinated histidines in light-harvesting complex II: assessment of charge transfer and electronic delocalization effect. AB - In a recent MAS NMR study, two types of histidine residues in the light harvesting complex II (LH2) of Rhodopseudomonas acidophila were resolved: Type 1 (neutral) and Type 2 (positively charged) (Alia et al. J. Am. Chem. Soc. ). The isotropic (13)C shifts of histidines coordinating to B850 BChl a are similar to fully positively charged histidine, while the (15)N shift anisotropy shows a predominantly neutral character. In addition the possibility that the ring currents are quenched by overlap in the superstructure of the complete ring of 18 B850 molecules in the LH2 complex could not be excluded. In the present work, by using two-dimensional heteronuclear ((1)H-(13)C) dipolar correlation spectroscopy with phase-modulated Lee-Goldburg homonuclear (1)H decoupling applied during the t(1) period, a clear and unambiguous assignment of the protons of histidine interacting with the magnesium of a BChl a molecule is obtained and a significant ring current effect from B850 on the coordinating histidine is resolved. Using the ring current shift on (1)H, we refine the (13)C chemical shift assignment of the coordinating histidine and clearly distinguish the electronic structure of coordinating histidines from that of fully positively charged histidine. The DFT calculations corroborate that the coordinating histidines carry approximately 0.2 electronic equivalent of positive charge in LH2. In addition, the data indicate that the ground state electronic structures of individual BChl a /His complexes is largely independent of supermolecular pi interactions in the assembly of 18 B850 ring in LH2. PMID- 14755160 TI - An isotope labeling strategy for methyl TROSY spectroscopy. AB - Recently we have shown that HMQC spectra of protonated methyl groups in high molecular weight, highly deuterated proteins have large enhancements in sensitivity and resolution relative to HSQC-generated data sets. These enhancements derive from a TROSY effect in which complete cancellation of intra methyl (1)H-(1)H and (1)H-(13)C dipolar interactions occurs for 50% of the signal in the case of HMQC, so long as the methyl is attached to a molecule tumbling in the macromolecular limit (Tugarinov, V., Hwang, P.M., Ollerenshaw, J.E., Kay, L.E. J. Am. Chem. Soc. (2003) 125, 10420-10428; Ollerenshaw, J.E., Tugarinov, V. and Kay, L.E. Magn. Reson. Chem. (2003) 41, 843-852. The first demonstration of this effect was made for isoleucine delta1 methyl groups in a highly deuterated 82 kDa protein, malate synthase G. As with (1)H-(15)N TROSY spectroscopy high levels of deuteration are critical for maximizing the TROSY effect. Here we show that excellent quality methyl TROSY spectra can be recorded on U-[(2)H] Iledelta1 [(13)CH(3)] Leu,Val-[(13)CH(3)/(12)CD(3)] protein samples, significantly extending the number of probes available for structural and dynamic studies of high molecular weight systems. PMID- 14755161 TI - Evidence of slow motions by cross-correlated chemical shift modulation in deuterated and protonated proteins. AB - Cross-correlated fluctuations of isotropic chemical shifts can provide evidence for slow motions in biomolecules. Slow side-chain dynamics have been investigated in (15)N and (13)C enriched ubiquitin by monitoring the relaxation of C(alpha) C(beta) two-spin coherences (Frueh et al., 2001). This method, which had hitherto been demonstrated only for protonated ubiquitin, has now been applied to both protonated and deuterated proteins. Deuteration reduces the dipole-dipole contributions to the DD/DD cross-correlation, thus facilitating the observation of subtle effects due to cross-correlation of the fluctuations of the isotropic (13)C chemical shifts. The decays of double- and zero-quantum coherences are significantly slower in the deuterated protein than in the protonated sample. Slow motions are found both in loops and in secondary structure elements. PMID- 14755162 TI - Phosphorus-31 transverse relaxation rate measurements by NMR spectroscopy: insight into conformational exchange along the nucleic acid backbone. AB - Phosphorus ((31)P) NMR spectroscopy can provide important information about the dynamics of nucleic acids. In this communication, we propose an inversely detected (31)P transverse relaxation rate ( R (2)) measurement experiment. This experiment enables fast measurement of accurate (31)P transverse relaxation rates and provides the possibility to detect slow motions mapped by the phosphorus nuclei along the nucleic acid backbone. Dispersion curves show some (31)P nuclei experiencing chemical exchange in the millisecond time scale. Under the assumption of a two-state exchange process, the reduced lifetimes of the exchanging sites (tau(ex)) obtained are in accordance with base pair lifetime estimates deduced from imino proton exchange measurements. PMID- 14755163 TI - Triple resonance MAS NMR with (13C, 15N) labelled molecules: reduced dimensionality data acquisition via 13C-15N heteronuclear two-spin coherence transfer pathways. AB - A reduced dimensionality magic angle spinning solid-state NMR experimental protocol for obtaining chemical shift correlation spectra of dipolar coupled nuclei in uniformly ((13)C, (15)N) labelled biological systems is described and demonstrated. The method involves a mapping of the evolution frequencies of heteronuclear (13)C-(15)N zero- and double-quantum coherences. In comparison to a reduced dimensionality procedure involving the simultaneous incrementation of two single-quantum chemical shift evolution periods, the approach described here could be potentially advantageous for minimising the heat dissipated in the probe by high power (1)H decoupling in experiments requiring long t (1) acquisition times. PMID- 14755164 TI - Backbone resonance assignment of the L-arabinose binding protein in complex with D-galactose. PMID- 14755165 TI - Sequence-specific 1H, 13C, 15N resonance assignments and secondary structure of the carboxyterminal domain of the E. coli transcription factor NusA. PMID- 14755166 TI - 1H, 13C and 15N assignments of single-stranded DNA binding domains from the 70 kDa subunit of human replication protein A. PMID- 14755167 TI - The C-terminal domain of viral IAP associated factor (cVIAF) is a structural homologue of phosducin: resonance assignments and secondary structure of the C terminal domain of VIAF. PMID- 14755168 TI - Backbone 1H, 13C, and 15N assignments of a 42 kDa RecR homodimer. PMID- 14755169 TI - Assignment of the 1H, 13C and 15N resonances of the LpxC deacetylase from Aquifex aeolicus in complex with the substrate-analog inhibitor TU-514. PMID- 14755170 TI - Backbone resonance assignments of human adult hemoglobin in the carbonmonoxy form. PMID- 14755171 TI - A taxonomic study of species of Bothriocephalus Rudolphi, 1808 (Cestoda: Pseudophyllidea) from eels in Japan: morphological and molecular evidence for the occurrence of B. claviceps (Goeze, 1782) and confirmation of the validity of B. Japonicus Yamaguti, 1934. AB - A taxonomic study of specimens of Bothriocephalus from eels ( Anguilla spp.) in Japan has demonstrated the occurrence of two species, B. claviceps (Goeze, 1782) and B. japonicus Yamaguti, 1934. The former species is a parasite of eels ( A. anguilla and A. rostrata ) in the Holarctic Region and was recently reported from A. marmorata in Japan. The conspecificity of tapeworms newly found in an eel ( A. ? japonica ) from Lake Biwa, central Japan, with B. claviceps has been confirmed by the great similarity of their ITS-2 gene sequences (similarity 95.3% and 95.2%). However, the sequences of worms identified as B. claviceps from A. marmorata differed considerably from those of B. claviceps from two populations of A. anguilla from Europe and the above-mentioned one from Japan (similarity 66.3%, 67.1% and 65.1 %, respectively), thus indicating that the former cestodes may have been misidentified. This assumption was confirmed by morphological evaluation of a voucher specimen from A. marmorata. The morphology of this cestode, as well as those from A. japonica from two localities in Japan (Lakes Biwa and Suwa), indicates their conspecificity with B. japonicus. The validity of this taxon has been confirmed on the basis of a re-examination of the type specimens. The two taxa, B. japonicus and B. claviceps, differ from each other in the shape and length of the scolex (619-730 microm in B. japonicus versus 1,180 2,100 microm in B. claviceps ), the relative position of the cirro-vaginal and uterine pores (opposite each other in relation to the median line of the body in B. japonicus versus tandem or slightly offset along the median line in the latter species), and the size of the eggs (41-52 x 28-35 microm in B. japonicus versus 50-70 x 31-43 microm in B. claviceps ). PMID- 14755173 TI - [Microphallidae (Digenea) from southern Africa, parasites of Charadrii (Aves). Second note]. AB - The authors present new geographical and diagnostic information for microphallids (Trematoda: Digenea) from the coast of Namibia (southern Africa): Maritrema eroliae Yamaguti, 1939 from Charadrius marginatus Vieillot; Odhneria odhneri Travassos, 1921 from Arenaria interpres L.; Microphallus bilobatus Cable, Connor & Balling, 1960 from C. marginatus; and Levinseniella propinqua Jagerskiold, 1907 from C. marginatus and A. interpres. These are new geographical and host records. The position and variability of the phallus (male copulatory organ) in M. bilobatus from Namibia and in the type-species from the Caribbean Sea are compared and illustrated. The genital atria of L. propinqua from Namibia and from Marcus Island (southwest Cape Province, Southern Africa) are illustrated and compared. This species appears to be cosmopolitan. PMID- 14755172 TI - Redescription of Electrotaenia malopteruri (Fritsch, 1886) (Cestoda: Proteocephalidae), a parasite of Malapterurus electricus (Siluriformes: Malapteruridae) from Egypt. AB - The proteocephalidean cestode Electrotaenia malopteruri (Fritsch, 1886) (Proteocephalidae: Gangesiinae), the type- and only species of Electrotaenia Nybelin, 1942 and specific to the electric catfish Malapterurus electricus Gmelin (Siluriformes: Malapteruridae), is redescribed on the basis of freshly collected material from the River Nile in Egypt. The validity of Electrotaenia is confirmed and some unique characters of this genus, observed in extensive material from different host specimens from Egypt, Sudan, Sierra Leone and Nigeria are first reported or described in detail. Such details include the internal morphology of a rostellum-like apical organ which is disc-shaped with a flat or slightly concave apex, the structure of the ovary which is follicular to reticulate, the structure of the cirrus-sac, the presence of a medio-dorsal band of muscle fibres, and the morphology of the vagina and eggs. PMID- 14755174 TI - On Branchiopodataenia n. g., parasitic in gulls, and its type-species, B. Anaticapicirra n. sp. (Cestoda: Hymenolepididae). AB - Branchiopodataenia n. g. is established for hymenolepidid cestodes characterised by the presence of a specific morphological feature, a latch-like structure in the copulatory part of the vagina. Their life-cycles involve branchiopod crustaceans (Branchiopoda) as intermediate hosts and gulls as definitive hosts. The type-species of the genus, B. anaticapicirra n. sp. from gulls of Chukotka (Chaun lowlands) and Alaska (River Yukon, Cape Barrow), is described. It is established that B. arctowskii (Jarecka & Ostas, 1984) n. comb. (originally Hymenolepis ) has a bipolar distribution and is not endemic to the Antarctic. A description of B. arctowskii from the northern hemisphere and the principal characters of other known species of this genus, B. gvozdevi (Maksimova, 1988) n. comb. (originally Wardium ), B. haldemani (Schiller, 1951) n. comb. (originally Hymenolepis ) and B. pacifica (Spassky & Jurpalova, 1968) n. comb. (originally Wardium ), are included. A key to the species of Branchiopodataenia is also presented. PMID- 14755175 TI - New quill mites of the family Syringophilidae (Acari: Cheyletoidea). AB - Five new species and two new genera belonging to the family Syringophilidae (Acari: Cheyletoidea) are described from birds that died in the Antwerp Zoo during their quarantine: Charadriaulobia vanelli n. g., n. sp. from Vanellus chilensis (Charadriiformes: Charadriidae) in Brazil; Fritschisyringophilus lonchurae n. g., n. sp. from Lonchura punctulata (Passeriformes: Estrildidae) in India; Mironovia coturnae n. sp. from Coturnix coturnix (Galliformes: Phasianidae) in Europe; Syringophiloidus daberti n. sp. from Passerina ciris (Passeriformes: Emberizidae) in Mexico; and S. serini n. sp. from Serinus mozambicus (Passeriformes: Fringillidae) in Central Africa. Charadriaulobia n. g. differs from the closely related Aulobia Kethley, 1970, in both sexes, by the divergent epimeres I; in females, by the absence of protuberances on the hypostomal apex and by the situation of the bases of setae l4 distinctly anterior the bases of setae d4. Fritschisyringophilus n. g. differs from the closely related Syringophiloidus Kethley, 1972, in both sexes, by the presence of setae vs ' on legs II, the absence of setae dT on legs III and IV; in females, by the presence of median hypostomal protuberances and by short setae l1, l2 and l3. The relationships between the Syringophilidae and their hosts are briefly discussed. A list of all known syringophilid genera and their distribution on bird families is provided. PMID- 14755176 TI - Ankistromeces mariae n. g., n. sp. (Digenea: Sanguinicolidae) from Meuschenia freycineti (Monacanthidae) off Tasmania. AB - Ankistromeces mariae n. g., n. sp. is described from Meuschenia freycineti (Monacanthidae), the six-spined leatherjacket, from off northern Tasmania. The new genus differs from the 21 other sanguinicolid genera in the combination of the anteriorly intercaecal and posteriorly post-caecal single testis, the presence of a cirrus-sac, the absence of an auxiliary external seminal vesicle, separate genital pores, the typically post-ovarian uterus and the H-shaped intestine. A. mariae is the first sanguinicolid to be reported from a monacanthid fish. PMID- 14755177 TI - Adjusting for covariates in variance components QTL linkage analysis. AB - Variance components modeling has emerged as a powerful method for quantitative trait loci (QTL) linkage analysis. However, the power to detect a gene of minor effect is low. Many complex traits are affected by environmental as well as genetic factors, and one strategy to increase power is to reduce nongenetic phenotypic variance by adjusting for environmental covariates. In this paper, we investigate the power of three approaches to covariate adjustment in variance components linkage analysis: (i) incorporate covariates in the means model, (ii) incorporate covariates in the covariance matrix, and (iii) perform analysis on residual statistics. These approaches are compared to an analysis without adjustment. The results show that in the absence of correlation between the covariate and the QTL effect, adjusting for covariates indeed increases power to detect an underlying QTL. As this correlation increases, however, the power decreases. In the presence of a causal association between QTL and covariates, not adjusting for covariates appeared to be more powerful. The three approaches for adjusting for covariate: residual statistics, the means model, and the covariance model, had equal power to detect a QTL. PMID- 14755178 TI - A simulation study concerning the effect of varying the residual phenotypic correlation on the power of bivariate quantitative trait loci linkage analysis. AB - The power of bivariate variance components (VC) linkage analysis is affected by the size and source of the phenotypic correlation between variables. In particular, several authors have suggested that the power to detect linkage is greatest when the quantitative trait locus (QTL) and residual sources of variation induce phenotypic covariation in opposite directions, and that this increase in power is greatest when unique environmental sources of variation induce covariation in the direction opposite to the QTL. The purpose of the present study was to investigate further the effect of varying the residual correlation between variables on the power to detect linkage in a bivariate variance components linkage analysis. Data were simulated for a biallelic QTL that pleiotropically influenced two variables. The power to detect linkage was calculated under a variety of situations in which the proportion of phenotypic covariance resulting from shared sources of variation and from unique sources of variation was varied. These simulations were performed for the case in which the QTL affected the two variables equally and also for the case in which the QTL made unequal contributions to each variable. Our results confirm that the power to detect QTLs in a bivariate test for linkage depends upon the size and source of the residual correlation between variables, being greatest when the QTL and unique environmental sources of variation induce phenotypic covariation in opposite directions. We also found that when the QTL affected the two variables unequally, the power to detect linkage increased markedly as the correlation between unique environmental sources of variation increased from 0.6 to 0.9. Similar results were obtained under a variety of genetic models, including when there were unequal allele frequencies and dominance at the QTL. We suggest that a promising strategy to increase the power to detect QTLs might be to collect data from variables where there is either good observational evidence (e.g., from multivariate structural equation modeling of twin data) or a sound theoretical argument that the QTL and environmental factors induce covariation in opposite directions. PMID- 14755179 TI - Epistasis in quantitative trait locus linkage analysis: interaction or main effect? AB - This paper explores a two-locus variance components model of quantitative trait locus (QTL) linkage for sib-pairs that incorporates epistasis. For a range of epistatic models the expected variance components and noncentrality parameter per sib-pair can be calculated, to indicate the power to detect epistasis. In QTL linkage analysis, additive and epistatic effects are in fact partially confounded; as a result, variance components under incorrect submodels can be distorted, with two main implications. First, the analysis of a single locus can in fact detect a QTL that has no main effect but interacts epistatically with another (unmeasured) locus. That is, single-locus approaches do not necessarily preclude the detection of purely epistatically interacting loci. Second, because the nonepistatic variance component estimates in submodels can partially absorb epistatic variance when it is not explicitly modeled, power to formally detect epistasis is low. PMID- 14755180 TI - Effect of winsorization on power and type 1 error of variance components and related methods of QTL detection. AB - Variance components analysis provides an efficient method for performing linkage analysis for quantitative traits. However, power and type 1 error of variance components-based likelihood ratio testing may be affected when phenotypic data are nonnormally distributed (especially with high values of kurtosis) and there is moderate to high correlation among the siblings. Winsorization can reduce the effect of outliers on statistical analyses. Here, we considered the effect of winsorization on variance components-based tests. We considered the likelihood ratio test (LRT), the Wald test, and some robust variance components tests. We compared these tests with Haseman-Elston least squares-based tests. We found that power to detect linkage is significantly increased after winsorization of the nonnormal phenotypes. Winsorization does not greatly diminish the type 1 error for the variance components-based tests for markedly nonnormal data. A robust version of the LRT that adjusts for sample kurtosis showed the best power for nonnormal data. Finally, phenotype winsorization of nonnormal data reduces the bias in estimation of the major gene variance component. PMID- 14755181 TI - Copulas in QTL mapping. AB - The standard variance components method for mapping quantitative trait loci is derived on the assumption of normality. Unsurprisingly, statistical tests based on this method do not perform so well if this assumption is not satisfied. We use the statistical concept of copulas to relax the assumption of normality and derive a test that can perform well under any distribution of the continuous trait. In particular, we discuss bivariate normal copulas in the context of sib pair studies. Our approach is illustrated by a linkage analysis of lipoprotein(a) levels, whose distribution is highly skewed. We demonstrate that the asymptotic critical levels of the test can still be calculated using the interval mapping approach. The new method can be extended to more general pedigrees and multivariate phenotypes in a similar way as the original variance components method. PMID- 14755182 TI - Exploring positional candidate genes: linkage conditional on measured genotype. AB - Variance component methods for linkage analysis of quantitative traits are now well established and have been applied with great success. Numerous QTLs influencing both normal variation between individuals and biomedically relevant quantitative risk factors have been localized in recent years. These findings have motivated recent methodological developments focused on the next step in the analytical process, moving beyond QTL localization by linkage to identifying specific genes and the functional polymorphisms in them that influence the phenotype of interest. In this paper, we describe one approach to exploring positional candidate genes in a region of linkage, linkage analysis conditional on measured genotype. We illustrate the properties of this method using simulated data from Genetic Analysis Workshops 10 and 12 and discuss example applications of this method to human quantitative trait data. PMID- 14755183 TI - Combined linkage and association tests in mx. AB - Statistical methods aimed at the detection of genes for quantitative traits suffer from two problems: (i) when a linkage approach is employed, relatively large sample sizes are usually required; and (ii) when an association approach is employed, effects of population stratification may blur genuine locus-trait associations. The variance components method proposed by Fulker et al. (1999) addressed both these problems; it is statistically powerful because it involves a combined analysis of linkage and association and can include information from multiplex families, which reduces the overall amount of necessary individual genotypes. In addition, it includes an explicit test for the presence of spurious association. After a brief illustration of the various ways in which population stratification may affect locus-trait associations, the implementation in Mx (Neale, 1997) of the method as proposed by Fulker et al. (1999) is discussed and illustrated. In addition, an extension to this method is proposed that allows the use of (variable) sibship sizes greater than two, the estimation of additive and dominance association effects, and the use of multiple alleles. These extensions can be implemented when parental genotypes are available or unavailable. PMID- 14755185 TI - Haplotype association analysis of discrete and continuous traits using mixture of regression models. AB - We present a regression-based method of haplotype association analysis for quantitative and dichotomous traits in samples consisting of unrelated individuals. The method takes account of uncertain phase by initially estimating haplotype frequencies and obtaining the posterior probabilities of all possible haplotype combinations in each individual, then using these as weights in a finite mixture of regression models. Using this method, different combinations of marker loci can be modeled, to find a parsimonious set of marker loci that are most predictive and therefore most likely to be closely associated with the a quantitative trait locus. The method has the additional advantage of being able to use individuals with some missing genotype data, by considering all possible genotypes at the missing markers. We have implemented this method using the SNPHAP and Mx programs and illustrated its use on published data on idiopathic generalized epilepsy. PMID- 14755184 TI - Modeling with measured genotypes: effects of the vitamin D receptor gene, age, and latent genetic and environmental factors on bone mineral density. AB - We use empirical data to demonstrate a recently proposed framework for including measured genotypes in structural equation models (Van den Oord and Snieder, 2002). The sample consisted of 227 MZ pairs, 197 DZ pairs, and 41 DZ twins without co-twin. The mean age of the female twins was 57 years. The analysis aimed at studying the effect of the vitamin D receptor (VDR) gene polymorphism (VDRGP) on bone mineral density (BMD). Whereas age affected the BMD measures via independent pathways, the effects of the VDRGP plus latent genetic and environmental factors were consistent with a common pathway model. VDRGP explained 0.3% to 0.5% of the variance. These analyses demonstrated that (i) regardless of how much is known about the gene and its products at a molecular level, meaningful questions can be addressed concerning how genes co-act with other variables at a "higher" phenotypic level; (ii) measured genotypes can be treated like any other variable so that the whole structural equation models framework can in principle directly be applied; and (iii) measured genotypes do not need to have large effects to study mechanisms underlying complex traits, and it may even be argued that particularly genes with small effects are better studied within the context of models with multiple variables. PMID- 14755186 TI - Advocate for patient care. PMID- 14755187 TI - Use of the 70-degree diamond burr in the management of complicated frontal sinus disease. AB - OBJECTIVES/HYPOTHESIS: Management of frontal sinus disease may require drill-out of bone in the frontal recess for access, ventilation, and drainage of the sinus cavity; removal of osteitic foci; or resection of neoplastic tissue. Technological advances, particularly burrs with angles of 70 degrees and stereotactic navigational imaging, offer new opportunities to provide access and minimize trauma. The preliminary study evaluates the safety and efficacy of such minimally invasive approaches. STUDY DESIGN: Retrospective review. METHODS: The authors describe the use of a 70-degree diamond burr in a series of 10 patients with complicated frontal sinus disease who underwent endoscopic frontal sinusotomy under stereotactic imaging guidance. RESULTS: The diagnoses consisted of frontal sinus mucocele (n = 4), chronic frontal sinusitis (n = 1), Pott's puffy tumor after frontoethmoid fracture (n = 1), and recurrent inverting papilloma (n = 4). Partial septectomy was required in 6 of 10 patients. No complications were attributable to the drill-out procedure, despite a pre existing frontoethmoid bony dehiscence in 6 of 10 patients. One patient had a CSF leak during removal of tumor from the skull base. One patient required revision frontal sinusotomy 10 months after the initial procedure, and another required further surgery for residual inverting papilloma on the medial orbital wall. All frontal sinusotomies were patent at last follow-up (mean period, 9.3 mo). CONCLUSION: Extended endoscopic frontal sinusotomy may be necessary in the management of complicated frontal sinus inflammatory disease and inverting papilloma. The 70-degree diamond burr is a safe and effective tool for access to the frontal recess. Complication rates appear to be similar to those for other extended frontal sinusotomy approaches. PMID- 14755188 TI - Different endoscopic surgical strategies in the management of inverted papilloma of the sinonasal tract: experience with 47 patients. AB - OBJECTIVE: To demonstrate the potentials and limitations of three different endoscopic procedures employed for treatment of inverted papilloma (IP) of the sinonasal tract. STUDY DESIGN: Retrospective analysis of a cohort of patients treated at two University hospitals. METHODS: From January 1992 to June 2000, 47 patients with IP underwent endoscopic resection. Preoperative workup included multiple biopsies of the lesion and imaging evaluation by computed tomography or magnetic resonance imaging. Massive skull base erosion, intradural or intraorbital extension, extensive involvement of the frontal sinus, abundant scar tissue caused by previous surgery, or the concomitant presence of squamous cell carcinoma were considered absolute contraindications for a purely endoscopic approach. Three types of resection were used: ethmoidectomy with wide antrostomy and sphenoidotomy (type 1) for IPs confined to the middle meatus, medial maxillectomy with ethmoidectomy and sphenoidotomy (type 2) for IPs partially invading the maxillary sinus, and a Sturmann-Canfield operation (type 3) for IPs involving the mucosa of the alveolar recess or of the anterolateral corner of the maxillary sinus. All patients were followed by periodic endoscopic evaluations. RESULTS: Type 1, 2, and 3 resections were performed in 26, 15, and 6 patients, respectively. No recurrences were observed after a mean follow-up of 55 (range 30 132) months. One patient, who underwent a type 2 resection, developed a stenosis of the lacrimal pathways requiring endoscopic dacryocystorhinostomy. CONCLUSIONS: Our experience confirms that endoscopic surgery is an effective and safe method of treatment for most IPs. The availability of different endoscopic techniques allows the entity of the dissection to be modulated in relation to the extent of disease. Strict application of selection criteria, meticulous use of subperiosteal dissection in the involved areas, and regular follow-up evaluation are key elements for success. PMID- 14755189 TI - Comparison of topical medication delivery systems after sinus surgery. AB - OBJECTIVES: To compare the distribution patterns of topical medication delivery systems in the sinonasal region and upper respiratory tract after functional endoscopic sinus surgery. STUDY DESIGN: Prospective descriptive evaluation. METHODS: Four topical delivery systems (spray bottle, atomizer, nebulizer, and bulb syringe) were studied. Using a dye solution as a marker, we independently applied the four topical delivery systems to a population of patients with chronic rhinosinusitis who had undergone functional endoscopic sinus surgery. The anatomic distributions were videotaped using flexible fiberoptic endoscopy. Three blinded observers independently rated the anatomic distribution of dye using a 4 point scale. Statistical analysis was performed using analysis of variance (ANOVA) and Dunn posttesting. RESULTS: Seven participants completed the study. All participants had undergone bilateral maxillary antrostomies, bilateral total ethmoidectomies, and bilateral sphenoidotomies. Five sinonasal sites and the larynx were evaluated for dye deposition. Interobserver agreement reached 95.6%. There was no statistical difference between the atomizer and spray bottle. The bulb syringe was statistically superior to the nebulizer in all sinonasal sites and statistically superior to the atomizer and spray bottle in the ethmoidal region. Dye was rarely seen within the larynx. CONCLUSIONS: The delivery systems tested were shown to have significant differences in their capability to place dye in specific sinonasal areas. Because topical medications are commonly administered to postoperative patients, these differences may have important clinical implications. PMID- 14755190 TI - The effects of upper airway surgery for obstructive sleep apnea on nasal continuous positive airway pressure settings. AB - OBJECTIVES/HYPOTHESIS: The objectives were to determine whether upper airway surgery lowers nasal continuous positive airway pressure (CPAP) settings for patients who require CPAP postoperatively for unresolved obstructive sleep apnea and to assess CPAP tolerability after upper airway surgery. STUDY DESIGN: Retrospective chart review. METHODS: Patients who underwent upper airway surgery with preoperative and postoperative polysomnography at the University of Alabama at Birmingham (Birmingham, AL) between 1995 and 2000 were the focus of the study. Upper airway surgery was defined as uvulopalatopharyngoplasty alone or in addition to septoplasty and turbinoplasty. Recommended CPAP settings were recorded from preoperative and postoperative polysomnography studies to determine whether CPAP settings were decreased following surgery. A response to surgery was defined as a decrease of the recommended CPAP setting by at least 1 cm of water. A telephone interview was conducted to determine whether upper airway surgery improved CPAP comfort. RESULTS: In 51.4% of the patients, CPAP settings were decreased following surgery. Continuous positive airway pressure settings were increased in 28.6% of patients and unchanged in 20%. Of the six patients who consistently used CPAP before and after surgery, four reported increased comfort postoperatively. CONCLUSION: Upper airway surgery does not predictably reduce CPAP settings in the patient who requires postoperative CPAP for unresolved obstructive sleep apnea. Upper airway surgery may improve CPAP tolerability. PMID- 14755191 TI - Outpatient endoscopic Zenker diverticulotomy. AB - OBJECTIVES/HYPOTHESIS: Staple-assisted, endoscopic Zenker diverticulotomy has been shown to decrease both cost of treatment and length of convalescence when compared with the standard open approach. Although the endoscopic technique is generally considered to be safe, the feasibility of outpatient endoscopic Zenker diverticulotomy has never been reported. STUDY DESIGN: Retrospective cohort study. METHODS: All endoscopic Zenker diverticulotomy procedures performed at Oregon Health and Science University (Portland, OR) between 1998 and 2002 were reviewed. The study group was limited to patients whose surgeries were planned on an outpatient basis. Medical charts were reviewed for indications, demographics, operative findings, complications, and resolution of symptoms. RESULTS: Of the 51 patients who underwent endoscopic, staple-assisted Zenker diverticulotomy, 40 were treated with the intent of outpatient management. Thirty-two cases were primary surgeries, and eight cases were revision treatments. The average patient age was 68 years (age range, 35-91 y), and the mean follow-up period was 5.9 months (range, 1-37 mo). Eight (20%) complications were noted in the study group. Intraoperative complications included 3 patients (7.5%) with mucosal disruptions. These patients underwent uneventful suture repair of small hypopharyngeal mucosal tears encountered intraoperatively and were temporarily observed in the hospital. One patient was admitted for postoperative urinary retention and another for fever and tachycardia 48 hours after surgery. One patient sustained a myocardial infarction. The remainder of patients were fed immediately and discharged home on the day of surgery. Two patients (5%) had iatrogenic tooth fracture. CONCLUSION: Appropriately selected patients undergoing endoscopic, staple-assisted Zenker diverticulotomy can be managed safely on an outpatient basis. Intraoperative complications, such as a mucosal disruption, warrant conversion to inpatient monitoring. PMID- 14755192 TI - Propofol decreases waste anesthetic gas exposure during pediatric bronchoscopy. AB - OBJECTIVE: This study compared the anesthetic gas exposure and operating conditions during insufflation anesthesia with halothane-alone versus halothane propofol in children undergoing direct laryngobronchoscopy. STUDY DESIGN: Forty six children were enrolled in this randomized prospective study, with institutional review board approval and informed consent. METHODS: All children were anesthetized by halothane mask induction and anesthesia was maintained using spontaneous ventilation with insufflation. No muscle relaxants or opioids were used. In the halothane group, halothane was titrated as needed. In the propofol group, halothane was decreased to 1% inspired concentration and the propofol was titrated as needed to maintain spontaneous ventilation and a still patient. Trace anesthetic gases, hemodynamic stability, and operating conditions were measured. RESULTS: The groups were similar in age, weight, and bronchoscopy time. There was significantly less gas exposure in the propofol group (25 +/- 33 parts per million) versus the halothane group (66 +/- 97 ppm; P <.02). There was a trend toward earlier emergence in the halothane group (33 +/- 13 minutes) versus the propofol group (41 +/- 17 minutes). Postoperative stridor was common, occurring in 30% of children. CONCLUSIONS: Insufflation anesthesia with spontaneous respiration provides excellent surgical conditions for laryngobronchoscopy. The addition of propofol resulted in fewer airway complications (P =.047). Although the addition of propofol significantly decreased anesthetic gas exposure in the operating room, both techniques resulted in operating room pollution that exceeded the maximum levels of 2 ppm per hour recommended by the US National Institute for Occupational Safety and Health (NIOSH). PMID- 14755193 TI - Vestibular nerve section versus intratympanic gentamicin for Meniere's disease. AB - OBJECTIVES/HYPOTHESIS: Vestibular nerve section and transtympanic gentamicin administration are procedures with proven efficacy in the treatment of vertigo associated with Meniere's disease refractory to medical management. Hearing loss is a known complication of each of these procedures; however, there has not been a report of hearing results of both treatments from a single institution. STUDY DESIGN: Retrospective review. METHODS: Review was made of 25 patients undergoing gentamicin injection and 39 patients undergoing vestibular nerve section for Meniere's disease. Rate of vertigo control and pretreatment and post-treatment pure-tone average values and speech discrimination scores were reported. RESULTS: The mean preoperative pure-tone average for patients having vestibular nerve section was 47.2 dB, with a speech discrimination score of 75.4%. In these patients, the postoperative pure-tone average was 49.1 dB and the speech discrimination score was 75%. Patients undergoing gentamicin injection had a mean pretreatment pure-tone average of 55.9 dB and a speech discrimination score of 62%. The post-treatment pure-tone average and speech discrimination score for the gentamicin group were 68.8 dB and 49.3%, respectively. Five of 25 patients (20%) in the gentamicin treatment group and 1 of 39 (3%) in the vestibular nerve section treatment group had an increase in bone-conduction threshold greater than 30 dB. The amount of postprocedure hearing loss was significantly greater in the gentamicin treatment group (P =.006). Control of vertigo was good to excellent in 95% of the patients treated with vestibular nerve section and in 80% of the patients treated with gentamicin. CONCLUSION: Although vestibular nerve section and transtympanic gentamicin are both acceptable treatment options for vertigo associated with Meniere's disease, gentamicin causes a higher level of hearing loss related to treatment and vestibular nerve section has higher vertigo control rates. PMID- 14755194 TI - Functional outcome after total parotidectomy reconstruction. AB - OBJECTIVES/HYPOTHESIS: The objective was to compare and contrast the functional and cosmetic outcomes of patients who underwent total parotidectomy with and without reconstruction using an inferiorly based sternocleidomastoid muscle flap. STUDY DESIGN: Retrospective review in the setting of a university medical center. METHODS: Twenty-four patients underwent a total parotidectomy. Fifteen patients had reconstruction with an inferiorly based sternocleidomastoid muscle flap, and nine patients had no reconstruction. Clinical examination was performed independently by two head and neck surgeons and one aesthetician to evaluate cosmetic outcome, presence of gustatory flushing or sweating, and return of facial nerve and greater auricular nerve function. RESULTS: With the mean follow up of 22 months, the group having reconstruction showed a better cosmetic outcome compared with the group without reconstruction. Objective testing for Frey syndrome demonstrated gustatory sweating in 20% of the group having reconstruction group versus 22% in the group without reconstruction. There was no difference in length of operation, hospital stay, or facial nerve function. Objective testing of facial sensation revealed that only 40% in the group having reconstruction had normal sensation to light touch compared with 78% in the group without reconstruction. CONCLUSION: The inferiorly based sternocleidomastoid muscle flap offers improved cosmetic results in patients undergoing total parotidectomy. However, there is a decreased return of greater auricular nerve function, probably attributable to relocation of the nerve stump anteriorly. Benefit was not seen in prevention of Frey syndrome measured objectively; however, the group having reconstruction had fewer clinical symptoms of gustatory sweating or flushing. PMID- 14755195 TI - The diagnostic utility of computed tomography for preoperative localization in surgery for hyperparathyroidism. AB - OBJECTIVES/HYPOTHESIS: Successful unilateral or minimal-access parathyroid exploration and reoperative surgery of the parathyroid glands requires accurate preoperative localization of parathyroid disease. Although ultrasound and nuclear imaging techniques have an established role in this regard, the use of computed tomography (CT) for parathyroid exploration is not well understood. The purpose of the present study was to better define the diagnostic utility of CT in preoperative localization of the abnormal gland in surgery for hyperparathyroidism. STUDY DESIGN: Retrospective cohort study. METHODS: All parathyroid explorations performed at Oregon Health and Science University (Portland, OR) between 2000 and 2002 were reviewed. The study group was limited to patients with hyperparathyroidism in whom localization failed preoperatively using ultrasound and/or sestamibi scanning and subsequent investigation using CT imaging was performed. Operative, pathological, and imaging reports were then analyzed to assess the accuracy of CT imaging for localizing parathyroid disease. RESULTS: Twenty-two patients with hyperparathyroidism were investigated preoperatively using CT imaging. Parathyroid exploration was successful in all but one patient, leaving 21 patients in all in the cohort. The majority of cases (67%) were reoperative, and all patients had previously undergone inconclusive ultrasound and/or sestamibi scanning. Computed tomography correctly localized parathyroid disease in 18 (86%) of the 21 patients who underwent successful extirpation of parathyroid disease and was able to identify abnormal glands with equal utility in the neck and the chest. CONCLUSION: When ultrasound or sestamibi are unsuccessful, CT imaging can provide valuable preoperative localizing information before surgery for hyperparathyroidism, particularly in patients with recurrent or persistent disease. PMID- 14755196 TI - Role of the physician versus the patient in the detection of recurrent disease following treatment for head and neck cancer. AB - OBJECTIVES/HYPOTHESIS: Objective evidence supporting current National Comprehensive Cancer Network guidelines regarding surveillance of patients treated for head and neck cancer is presently lacking. The study examines the relative role of current surveillance methods on disease detection in this patient population. STUDY DESIGN: Prospective nonrandomized study. METHODS: Clinical information was prospectively collected in a standardized format during 3645 encounters with patients with head and neck cancer over an 18-month period. Data pertaining to visit history, symptom history, patient findings, physician findings, and disease status for each encounter were reviewed. RESULTS: Of 3645 visits, disease recurrence or new primary tumor was documented in 180 encounters (5%). Salvage therapy was thought to be feasible in at least 65% of cases. Of these 180 recurrences or new primaries, there were 142 patients (79%) who had identified new symptoms or physical findings, or both, before the physician's examination. Most commonly reported was the presence of a neck mass (38%), progressive pain (27%), or other visible lesion or ulcer (14%). Patients with recurrence represented nearly 40% of all patients reporting new symptoms or findings (142 of 367). Conversely, recurrence was rare in the absence of reported symptoms or findings (1.2%). Surprisingly, despite patients reporting new symptoms or findings, physician evaluation most commonly occurred at the patient's routine surveillance visit rather than an earlier time point (104 of 142 [73%]). CONCLUSION: Self-diagnosis of recurrent or new primary disease is extremely common by virtue of symptoms or findings noted by patients before interaction with the clinician. However, presence of symptoms or findings did not motivate the patients to seek earlier medical attention. In the absence of such symptoms, physician diagnosis of recurrence is uncommon. Given the significant social and economic impact involved in surveillance of patients with head and neck cancer, further prospective study to optimize the method and frequency of this type of clinical activity is warranted and planned. PMID- 14755197 TI - Endoscopic vertical partial laryngectomy. AB - OBJECTIVE: To explain the significant difference between microlaryngoscopy with cordectomy and endoscopic vertical partial laryngectomy (EVPL), to describe the efficacy of EVPL on T1b and T2 glottic squamous cell carcinoma, and to evaluate EVPL with postoperative irradiation in T2 glottic cancer with impaired true vocal cord mobility. STUDY DESIGN: Retrospective review. METHODS: Twenty-six patients seen at the University of Utah Health Science Center between 1987 and 2000 with bilateral T1 (T1b) or T2 squamous cell carcinoma of the glottic larynx underwent EVPL. T2 cancers were classified as follows: a = unilateral disease, b = bilateral disease; i = impaired mobility. T1b and T2a glottic cancer patients received surgery alone, whereas impaired mobility patients (T2ai + T2bi) patients received surgery followed by planned postoperative irradiation. Patients were assessed for primary site control, perioperative and long-term complications, and ultimate cancer control. RESULTS Survival in the total group was 88.5%, with local control at 92.3%. The two recurrent patients were salvaged by total laryngectomy. For the whole group, anterior commissure involvement was present in 57.7% (15 of 26). Thirteen T2 (5 T2ai + 8 T2bi) carcinoma patients underwent combined therapy, with 8 (61.5%) of these patients having anterior commissure involvement. Two of these patients were upstaged at surgery, one to T3 and one to T4. Local control was 84.5%. Thirteen patients were treated by surgery only, with five of these patients having failed previous irradiation. Survival was 92.3% and local control 100%. This group included two T2bi patients, two patients upstaged to T4 on the basis of extension beyond the subglottis to the anterior wall of the trachea, 3 T2b, and 6 T2a patients. Anterior commissure involvement was seen in 7 (53.8%) of these patients. CONCLUSIONS: EVPL alone controlled all T1b and T2a glottic cancer patients, even in the presence of greater than 50% anterior commissure involvement. The significant difference between EVPL and classical microlaryngoscopy with cordectomy was carefully described. EVPL with planned postoperative irradiation resulted in an 85% local control rate in clinically staged T2ai and T2bi cancer patients, including the three upstaged patients. PMID- 14755198 TI - Current applications of microarrays in head and neck cancer research. AB - OBJECTIVES/HYPOTHESIS: The objective was to introduce microarray technology and its applications in cancer research to the head and neck clinician. STUDY DESIGN: Literature review combined with methodology and examples from the authors' experiences with microarray analysis of tumors of the head and neck. METHODS: Search of literature and the authors' experience was made for technical details, alternative methods of data analysis, available bioinformatics tools, and applications of microarrays in cancer research. RESULTS: Microarrays allow the simultaneous analysis of the expression of thousands of genes. The use of a well developed microarray study design leads to informative results. There are various bioinformatics resources widely available to aid in the analysis of microarray data. However, there is not yet a gold standard for analysis because this methodology is still evolving. CONCLUSION: Microarray studies may allow researchers to identify genetic changes relevant to diagnosis and prognosis in patients with head and neck cancer. Although still relatively new, this powerful methodology has immense potential to aid in understanding of the genetic changes that are important in head and neck cancer. PMID- 14755199 TI - Myxoma of the lateral skull base: clinical features and management. AB - OBJECTIVE: To evaluate the clinical features and incidence of true myxomas affecting the lateral skull base. MATERIAL AND METHODS: Illustrative case report and review of the relevant literature. Included are all reports of myxomas according to the histopathologic definition of Stout. RESULTS: Despite nine case reports, only four patients matched the histopathologic criteria of a true myxoma growing in the lateral skull base. Facial palsy occurred in three adults with additional otalgia in one and progressive hearing loss (HL) in another patient. Our patient initially presented with signs of recurrent otitis media. The mastoid cavity was involved in all patients. CONCLUSION: Differential diagnosis of lesions occurring in the lateral skull base should include true myxoma. Correct diagnosis should be based on immunohistopathology to rule out malignant tumors. Radical surgery is the treatment of choice, but surgeons should respect vital structures and esthetic consequences. PMID- 14755200 TI - Nasal cerebrospinal fluid leaks: critical review and surgical considerations. AB - Otolaryngologists have assumed a major role in the evaluation and management of anterior skull base defects that result in cerebrospinal fluid (CSF) leaks and meningoencephaloceles. To achieve the best possible results for patients with CSF leaks, a thorough understanding of the underlying pathophysiology and a critical analysis of management principles and treatment options is essential. Surgical and medical management of these patients is highly individualized and depends on a number of factors, including etiology, anatomic site, patient age, and underlying intracranial pressure. This review article will highlight the history, physiology, pathophysiology, diagnosis, surgical techniques, and postoperative care relevant to nasal CSF leaks and encephaloceles. PMID- 14755201 TI - Use of sodium fluorescein solution for detection of cerebrospinal fluid fistulas: an analysis of 420 administrations and reported complications in Europe and the United States. AB - BACKGROUND: Localization of dural fistulas in the region of the anterior or lateral skull base may be difficult. For many years, a sodium fluorescein solution of 0.5 to 5% (2.5-50 mg) has been administered intrathecally by way of the lumbar space. However, fluorescein is not commercially available for this stated purpose in either Germany or the United States. METHODS: Retrospectively, 420 fluorescein applications by the authors were retrospectively analyzed. Under the Freedom of Information Act, the United States Federal Drug Administration and the manufactures of fluorescein were queried for adverse reaction reports. RESULTS: Four hundred twenty fluorescein applications in 305 patients could be evaluated. Mean age of recipients was 46.9 years, ranging from 1 to 82 years. At a concentration of 5% fluorescein, 26 patients on the day of surgery and 69, 37, 34, and 14 patients on days 2 through 4 suffered from minor side effects that may or may not have been related to this drug. Two of these patients had grand mal seizures, which were attributable to simultaneous intrathecal application of contrast medium. All other side effects were thought to be the result of a postspinal headache and related lumbar puncture. At a concentration of 0.5%, the intraoperative intrathecal administration of 0.5 to 2 mL of fluorescein followed by 4 to 5 days of lumbar drainage resulted in some degree of spinal headache without other complications. In both groups, no patient had sequelae longer than 4 weeks. An additional seven complications were reported to the Federal Drug Administration and the fluorescein manufacturers at doses of 100 to 700 mg. CONCLUSIONS: Complications from intrathecal application of fluorescein appears to be dose dependent. At concentrations of 5%, or preferably lower, side effects are transient. A grand mal seizure can be minimized when following the general cautions of lumbar puncture and dose. The patient should be supervised for 24 hours. A written informed consent from patients for use of fluorescein is recommended. PMID- 14755202 TI - Experimental sinusitis in a rhinogenic model. AB - OBJECTIVES/HYPOTHESIS: The objectives were to determine the optimal sinusitis induction period and to examine microbiological and histopathological changes of sinusitis recovery stage in a rhinogenic sinusitis model. METHODS: A synthetic sponge was inserted into the right-side nasal cavities of rabbits. The sponge was impregnated with a Streptococcus pneumoniae strain in group 1 and with sterile saline solution in groups 2 and 3. After the fourth day of sponge insertion, sinuses were examined by coronal computed tomography scans at two-day intervals until any radiological evidence of sinusitis was observed. When sinusitis was detected radiologically, five rabbits each from groups 1 and 2 were killed for histological examination. To determine the recovery period of sinusitis, sponges were removed from the rest of the rabbits in groups 1 and 2. Rabbits were selected randomly and killed on the 15th and the 30th days of the recovery period, immediately after radiological examinations. Group 3 was considered a sham group. RESULTS: Sinusitis induction was performed in all rabbits in groups 1 and 2 until the 8th day. After the sponges were removed, inflammation persisted until the 30th day of the study. CONCLUSION: In a rhinogenic sinusitis model, although histological features of sinusitis were demonstrated, further studies are required to standardize this model and to examine whether or not the studied bacterial strain spreads from nasal cavity into sinus. PMID- 14755203 TI - Pathology of the olfactory mucosa: implications for the treatment of olfactory dysfunction. AB - OBJECTIVE: The pathology of the olfactory mucosa is poorly understood; however, most cases of hyposmia and anosmia appear to be associated with a decline in the number of functioning mature olfactory sensory neurons (OSNs). Under normal conditions, OSNs undergo apoptotic cell death at a baseline rate likely secondary to their exposed location in the nose. Regeneration of mature OSNs from precursors in the epithelium allows the animal to maintain an adequate number of neurons necessary for olfactory sensation. In many cases of olfactory dysfunction, this balance is apparently disturbed, with a net loss of OSNs. The current study will examine normal and diseased olfactory tissue for the presence of data demonstrating that the preferred mechanism of OSN cell death is apoptotic in both health and disease. The potential therapeutic implications will be discussed. STUDY DESIGN: Histologic analysis of human and animal olfactory tissue. METHODS: Normal and diseased human and animal olfactory mucosa were assessed for immunohistochemical evidence of apoptosis. RESULTS: Increased activity of the apoptotic effector enzyme caspase-3 was demonstrated in diseased olfactory mucosa in comparison with normal controls. CONCLUSION: These results indicate that a common pathway may mediate OSN cell death from a diverse set of pathologic insults including aging, trauma, and sinusitis. Interference with this pathway of cell death is currently the subject of intense pharmacotherapeutic research for the management of stroke and meningitis. These drugs may ultimately prove useful in the treatment of clinical olfactory dysfunction. PMID- 14755204 TI - Effect of clarithromycin on nuclear factor-kappa B and transforming growth factor beta in chronic rhinosinusitis. AB - OBJECTIVES: Long-term, low-dose macrolide therapy is effective in the treatment of chronic rhinosinusitis. It is believed that macrolide antibiotics produce this benefit through an anti-inflammatory effect. In this study, the effect of clarithromycin treatment on the expression of transforming growth factor (TGF) beta and the key pro-inflammatory nuclear transcription factor, NF-kappa B, was examined in vitro and in vivo. STUDY DESIGN AND METHODS: In vitro: nasal mucosa was obtained from 10 patients with chronic sinusitis and was cultured for 24 hours in the presence of clarithromycin or control. Cellular expression of TGF beta and NF-kappa B was determined by immunohistochemistry. In vivo: 10 patients with chronic rhinosinusitis were treated for 3 months with clarithromycin. Nasal mucosal biopsies were taken pre- and posttreatment. Cellular expression of TGF beta and NF-kappa B was again determined by immunohistochemistry. RESULTS: Clarithromycin, when applied to nasal biopsies in vitro, reduced cellular expression of TGF-beta and NF-kappa B. Nasal biopsies taken before and after clarithromycin treatment showed no differences in cellular expression of NF-kappa B or TGF-beta. CONCLUSION: Clarithromycin can reduce cellular expression of TGF beta and NF-kappa B when applied in vitro, but its action during clinical therapy is less clear. Clarithromycin is capable of inhibiting pro-inflammatory cytokines in vitro, and reductions of TGF-beta and NF-kappa B may represent additional mechanisms by which macrolides reduce inflammation in chronic airway disease. Discrepancies between the actions of clarithromycin on nasal biopsies in vitro and after clinical therapy warrant further investigation. PMID- 14755205 TI - Juvenile angiofibromas: changing surgical concept over the last 20 years. AB - OBJECTIVES: Nasopharyngeal angiofibroma is a rare tumor. Several surgical approaches have been established, including transpalatal, lateral rhinotomy, craniofacial, and midface degloving procedures. More recently, less invasive endonasal approaches have been used. In a retrospective study, we analyzed the outcome of surgical treatment for angiofibroma to determine whether an endonasal approach was as effective as other, more invasive techniques for removing the tumors. METHODS: Thirty patients with histologically confirmed angiofibromas were treated by surgical removal at two institutions during a 20-year period. Tumors were staged according to size and extension on the basis of imaging and surgical findings using the Fisch classification. Specific analysis of the operative approaches and recurrence rate by stage were included. Recurrent or residual disease was handled by surgical excision. RESULTS: The number of patients treated increased during the last 10 years in both institutions, but no major changes in distribution of tumor stages were observed. In the most recent years, the endonasal approach has been favored. All 30 patients remain alive and free of disease. CONCLUSIONS: During the last 12 years, we have undergone a marked shift toward endonasal procedures while the tumor stages of the patients treated remained the same. Our results demonstrate that angiofibromas may be managed successfully using less invasive techniques, thereby reducing morbidity, without increasing the chance of recurrence. PMID- 14755206 TI - The effect of electrocautery versus cold scalpel technique on the incidence of early postoperative tonsillar pillar dehiscence after uvulopalatopharyngoplasty with tonsillectomy. AB - OBJECTIVES/HYPOTHESIS: The objective was to determine the effect of electrocautery versus cold scalpel technique on the incidence of early postoperative tonsillar pillar dehiscence after uvulopalatopharyngoplasty with tonsillectomy. STUDY DESIGN: Prospective, randomized, matched-pair trial of patients undergoing uvulopalatopharyngoplasty with tonsillectomy. METHODS: Patients undergoing uvulopalatopharyngoplasty with tonsillectomy were randomly assigned to have the procedure performed with cold scalpel technique or with electrocautery. Thirty-three patients were enrolled. Twenty-seven patients returned for postoperative evaluation. RESULTS: The overall incidence of tonsillar pillar dehiscence was 38.8%. The incidence of dehiscence was 44.4% in the electrocautery group and 33.3% in the cold dissection group. CONCLUSION: Preliminary data suggest that the use of electrocautery is not associated with an increase incidence of wound dehiscence compared with cold dissection. Tonsillar pillar dehiscence rate is high, regardless of dissection method. PMID- 14755207 TI - Complications of microdebrider-assisted powered intracapsular tonsillectomy and adenoidectomy. AB - OBJECTIVES: To study complications of powered intracapsular tonsillectomy and adenoidectomy (PITA) in pediatric patients with obstructive sleep apnea (OSA). STUDY DESIGN: Retrospective chart review and long-term follow-up in office or by telephone interview. METHODS: We studied 278 patients who underwent PITA between September 2000 and October 2002. Outcome measures were postoperative bleeding, velopharyngeal insufficiency, need for hospital readmission, tonsil regrowth, and return of snoring or sleep apnea symptoms. RESULTS: All 278 children treated by PITA had immediate resolution of symptoms of OSA. Complications were noted in 11 patients (3.9%). Nine patients (3.2%) experienced tonsil regrowth with snoring, two of whom evolved to a return of OSA that was definitively managed by means of a complete tonsillectomy. Two patients (0.7%) had self-limited bleeding. None of the patients developed persistent velopharyngeal insufficiency or required hospital readmission. CONCLUSIONS: Microdebrider-assisted PITA is a safe and effective alternative for children otherwise treated with traditional tonsillectomy for symptoms of OSA due to adenotonsillar hypertrophy. This series suggests a 3.9% overall rate of complications, with the most common noted as tonsillar regrowth without recurrence of OSA. Prospective trials with longer follow-up may define higher complication rates. PMID- 14755208 TI - Effects of Tisseel and FloSeal on primary ischemic time in a rat fasciocutaneous free flap model. AB - OBJECTIVES: Free flaps are the technique of choice for reconstruction of defects resulting from extirpation of tumors of the head and neck. Advances in microsurgical technique have resulted in success rates of greater than 95%. Numerous intraoperative factors, ranging from technical issues to topically applied agents, can complicate the outcome of microsurgical free tissue transfer. Synthetic tissue adhesives and hemostatic agents are playing an ever-increasing role in reconstructive surgery. The safety of these factors in free flap surgery has not been ascertained. STUDY DESIGN: Animal Care Committee live rat model. METHODS: Male Sprague-Dawley rats were divided into three groups: group I, Control; group 2, FloSeal; group 3, Tisseel. In each group, a 3 x 6 cm ventral fasciocutaneous groin flap based on the left superficial epigastric artery was elevated and the experimental material applied beneath the flap and around the flap pedicle prior to suturing of the flap back to the wound bed. The experimental materials consisted of 0.2 mL saline in the control group, 0.5 mL FloSeal, and 0.2 mL Tisseel. In phase I of this study, the effect of each treatment on flap survival was assessed by survival at postoperative day 4. In phase II of the study, the effects of these agents on ischemic tolerance was investigated. Five rats in each treatment group were exposed to ischemic times of 6, 8, 10, and 12 hours. Survival of the flap was assessed 7 days after reversal of the ischemia. Probit curves and the critical ischemic time (CIT50) were calculated. RESULTS: All flaps survived the 2-hour period of ischemia and were viable at postoperative day 4. Flap survival from group 1 (Control), group 2 (FloSeal), and group 3 (Tisseel) at the various ischemic times was as follows: at 6 hours, 80%, 80%, and 80%, respectively; at 8 hours, 80%, 80%, 60%; at 10 hours, 60%, 33%, 40%; at 12 hours, 20%, 20%, 0%. The CIT50 for the Control, FloSeal, and Tisseel groups was 9.4, 9.0, and 7.0 hours, respectively.CONCLUSIONS FloSeal, a thrombin-based hemostatic agent, and Tisseel, a fibrin glue, displayed no adverse effect on flap survival in this model. PMID- 14755209 TI - Optimum tension for partial ossicular replacement prosthesis reconstruction in the human middle ear. AB - OBJECTIVE: Hearing results from ossiculoplasty are unpredictable. There are many potentially modifiable parameters. One parameter that has not been adequately investigated in the past is the effect of tension on the mechanical functioning of the prosthesis. Our goal was to investigate this parameter further, with the hypothesis that the mechanical functioning of partial ossicular replacement prostheses (PORP) from the stapes head to the eardrum will be affected by the tension that they are placed under. METHODS: Fresh temporal bones were used to reconstruct a missing incus defect with a PORP-type prosthesis. Three different lengths of PORP were used, and the stapes vibrations were measured with a laser Doppler vibrometer using a calibrated standard sound in the ear canal. Eight temporal bones were used. RESULTS: Tension had a very significant effect on stapes vibration. In general, loose prostheses resulted in the best overall vibration transmission. The effects were most marked at the lower frequencies. There was a slight advantage to tight prostheses in the higher frequencies, but much less than the decrement in lower frequencies with tight prostheses. CONCLUSION: In ossicular reconstruction, best stapes vibration results in our model are achieved by shorter prostheses, which result in lower tension. PMID- 14755210 TI - Cochlear implantation in children with congenital inner ear malformations. AB - OBJECTIVE/HYPOTHESIS: To assess the audiologic and surgical outcomes for pediatric cochlear implant patients with inner ear malformations. STUDY DESIGN: Retrospective review of 315 pediatric cochlear implant cases from 1994 to 2002. METHODS: Twenty-eight pediatric cochlear implant patients with known inner ear malformations determined on high-resolution computed tomography (HRCT) of the temporal bone were the subjects of review. Results of HRCT findings, intraoperative findings, postoperative complications, and objective measures of both closed- and open-set testing of speech perception were analyzed. RESULTS: Patients with the constellation of an incompletely partitioned (IP) cochlea, enlarged vestibular aqueduct (EVA), and a dilated vestibule (i.e., Mondini's malformation) as well as those with an isolated EVA or partial semicircular canal aplasia have relatively good levels of speech perception. Patients with total semicircular canal aplasia, isolated IP, cochlear hypoplasia, or common cavity demonstrated lower levels of performance. Poor performance may be related to associated developmental delays rather than labyrinthine anatomy alone. Complications of surgery were relatively limited. CONCLUSIONS: Cochlear implantation can be successfully performed in children with inner ear malformations. These children and their parents can expect significant auditory benefits from this intervention. The various types of inner ear malformations may have quite different prognoses for good auditory performance. PMID- 14755211 TI - Surgical management of complications after hearing aid fitting. AB - OBJECTIVES: Standard procedures for hearing aid fitting performed in accordance with established guidelines are well tolerated, safe, and effective. In this article, we present unusual complications after hearing aid fitting that required surgical management. METHODS: Four otologists at a major university center with a combined 65 years of experience performed a retrospective analysis of their surgical practice. Six patients were identified that had encountered severe complications from improper earmold fitting that required surgical intervention. RESULTS: One patient had a perforation of the tympanic membrane with earmold material found to have migrated into the middle ear cleft. The tympanic membrane healed spontaneously, resulting in persistent otalgia and a maximum air-bone gap. The earmold cast was successfully removed by means of a tympanomastoidectomy. Two patients with presbycusis and normal ear canals developed eardrum perforations and conductive hearing deficits. In both patients, earmold material was found partially occupying the middle ear cleft and removed by way of a transcanal approach. Three patients with prior canal wall down mastoidectomy defects and narrow external ear canals required microtoscopy under general anesthesia or canaloplasty for removal of impacted material. CONCLUSION: Proper fitting of hearing aids performed by well-trained medical professionals results in a very low incidence of significant complications. Perforation of the tympanic membrane with impaction of earmold material in the middle ear or mastoid bowl may occur and can be successfully managed by standard otologic surgical techniques. PMID- 14755212 TI - Long-term effects of cerebellar retraction in the microsurgical resection of vestibular schwannomas. AB - OBJECTIVE: To determine the long-term effects, if any, of the greater cerebellar retraction that is required for retrosigmoid vestibular schwannoma versus resection as compared with the minimal, if any, cerebellar retraction required for translabyrinthine versus resection. STUDY DESIGN: Retrospective case control. METHODS: All patients who underwent retrosigmoid versus resection between 1988 to 2000 by one surgeon were identified. These patients were contacted and asked to complete the Dizziness Handicap Inventory (DHI). These patients were then matched to patients who underwent translabyrinthine versus resection according to age at surgery, age at time of study, sex, and size of tumor. Total DHI and subcategory DHI scores were compared using the Student t test. RESULTS: Thirty-three patients underwent the retrosigmoid approach, and 27 were contacted for completion of the survey, with a 81.3% response rate. Forty-six patients who underwent translabyrinthine versus resection were matched to the retrosigmoid group. Thirty six were contacted and completed the survey for a 75% response rate. Twenty-seven patients that most closely matched the retrosigmoid group comprised the translabyrinthine group used for comparison. Minimal differences were seen between the two groups. Mean total DHI score for the retrosigmoid group was 17.6, which was not significantly different from the mean score of 16.8 seen in the translabyrinthine group (P =.888). When comparing small tumors (<1.5 cm), total DHI scores were not found to be significantly different (P =.859). We observed similar findings in those with intermediate-sized tumors (1.5-3 cm). CONCLUSION: The significantly greater cerebellar retraction that is necessary for retrosigmoid versus resection does not result in greater long-term balance and disability as compared with those patients who have undergone the translabyrinthine approach. PMID- 14755213 TI - Vertebrobasilar occlusive disorders presenting as sudden sensorineural hearing loss. AB - BACKGROUND: Isolated sudden sensorineural hearing loss (SSHL) has been rarely related to vertebrobasilar occlusive disorders (VBOD). This is an important issue for both neurologists and otolaryngologists, since the management and prognosis of this type of hearing loss widely differs from that of hearing loss from other causes. OBJECTIVES: To describe the clinical characteristics and report the incidence of SSHL related to VBOD. METHODS: Retrospective analysis of clinical charts from 333 patients admitted for SSHL in a large ear, nose, and throat emergency tertiary care center from 1999 to 2002. RESULTS: Four cases (1.2%) of VBOD as the unique cause of SSHL were diagnosed among 333 patients. The most typical features of these cases were the presence of one of the following characteristics: (1) bilateral SSHL, (2) associated occipital or posterior nuchal pain, and (3) the occurrence of delayed neurologic deficits. The underlying vascular disease affected the vertebral arteries: dissection in two cases and atherosclerosis in two other cases. The audiometric features of hearing loss were endocochlear in one case, of both types in one case, and unknown in two cases. Hearing recovered partially or completely. CONCLUSIONS: Our results confirm the low incidence of SSHL related to VBOD and show that the observation of endocochlear audiometric features cannot preclude a central cause in SSHL. The clinical presentation of our cases related to VBOD emphasize that a careful follow-up of any patient with SSHL is warranted and that the presence of nuchal pain suggestive of arterial dissection in younger subjects, a past history of atherosclerosis or embolism in older patients, or the occurrence of delayed associated neurologic symptoms should be considered with particular caution in this situation. PMID- 14755214 TI - Ebselen-mediated protection from single and repeated noise exposure in rat. AB - OBJECTIVES/HYPOTHESIS: Ebselen, a glutathione peroxidase mimic, is a candidate compound for the prevention of noise-induced hearing loss. STUDY DESIGN: Single blinded, placebo-controlled study. METHODS: Methods included single and repeated noise exposures on F-344 female rats given oral or injected ebselen or vehicle before and after noise, evoked auditory brainstem responses using click and pure tone stimuli, light and fluorescence microscopy of cochleae stained with 4',6 Diamidino-2-phenylindole (DAPI) and fluorescein isothiocyanate-phalloidin, and statistical power determined by ANOVA. RESULTS: Auditory brainstem response indicated that ebselen provided significant protection from both temporary and permanent threshold shifts following single and repeated noise exposure. On average, three times more outer hair cells were lost in control versus ebselen treated animals. CONCLUSION: Ebselen reduces noise-induced hearing loss in rats. PMID- 14755215 TI - Vestibular evoked myogenic potentials and glycerol testing. AB - OBJECTIVES/HYPOTHESIS: The objective was to evaluate dizziness as the first symptom of endolymphatic hydrops, which could provide valuable information on the initial stages of endolymphatic hydrops development. STUDY DESIGN: The present investigation was specifically designed to establish whether a combination of the traditional glycerol test and the vestibular evoked myogenic potential (VEMP) glycerol test is capable of diagnosing endolymphatic hydrops early and to identify potential Meniere's disease. The study was limited to patients who complained of dizziness as their only vestibular symptom. METHODS: Thirty-eight consecutive patients with dizziness who had received no treatment were investigated. Each patient underwent glycerol testing measured with both conventional pure-tone audiometry and vestibular evoked myogenic potential testing. For the vestibular evoked myogenic potential glycerol test, an increase of more than 20% in both latency and amplitude after glycerol intake was considered an improvement. RESULTS In one patient, the reappearance of the vestibular evoked myogenic potentials was observed. In another six patients, there was an increase in the vestibular evoked myogenic potential amplitudes, unilaterally in four cases and bilaterally in two. One patient showed a bilateral improvement in amplitude and an unilateral amelioration in latency. CONCLUSION: The assumption of saccular dilation as a sign of early Meniere's disease was not supported by the literature, which considered saccular hydrops as the final progression of the dilation of the endolymphatic compartments into the cochlea. These data seem to indicate that in our patients a saccular dysfunction rather than an early saccular hydrops has been documented, confirming vestibular evoked myogenic potential testing as an intriguing diagnostic option for a lesion of this structure. The vestibular evoked myogenic potential glycerol test proved capable of identifying dysfunctions of the saccule that were not otherwise detectable by the routine methods. Therefore, the vestibular evoked myogenic potential glycerol test should be considered as an additional method of diagnosis in patients with vertigo or dizziness. PMID- 14755216 TI - Audiovestibular findings in patients with mitochondrial A1555G mutation. AB - OBJECTIVE: The aims of this study were to explore the prevalence of the A1555G mutation among a group of Japanese patients and to assess the pathophysiology of the hearing impairment associated with the mutation. STUDY DESIGN: Genetic study and retrospective chart review. METHODS: We screened for the mitochondrial DNA A1555G mutation in 138 unrelated Japanese deaf patients, including 63 sporadic cases and 75 familial cases with different patterns of inheritance. When available, patients carrying the mutation received audiovestibular examinations, including speech audiometry, distortion-product otoacoustic emission (DPOAE) testing, electrocochleography (ECochG), and electronystagmography. RESULTS: One of 63 sporadic cases (1.6%) and 6 of 75 familial cases (8.0%) carried the A1555G mutation. Patients with the mutation and a familial history included two with autosomal recessive inheritance and four with maternal inheritance. In addition, two of six patients (33.3%) presenting with aminoglycoside-induced sensorineural hearing loss (SNHL) were associated with the A1555G mutation. All but one of the patients carrying the mutation showed high-frequency SNHL. Distortion-product levels of DPOAE were reduced to the noise levels, suggesting the A1555G mutation caused cochlear deafness. Cochlear microphonics in ECochG showed elevation of the detection thresholds and corresponding audiometric thresholds. The ECochG data implied that patients with high-frequency SNHL had impairment of the cochlear hair cells that was most severe toward the basal turn. The electronystagmographic findings indicated no apparent vestibular dysfunction. CONCLUSIONS: Screening for the A1555G mutation, even in patients with idiopathic bilateral SNHL, likely would be useful for preventing further development and/or acceleration of the deafness. PMID- 14755218 TI - Clinical predictors of difficult laryngeal exposure. AB - PURPOSE: Although difficult laryngeal exposure (DLE) is a common problem encountered after procedures using a rigid laryngoscope, to date, no anticipatory physical indicators have been formally noted as reliable predictors of DLE. The purpose of this paper is to identify useful and reliable indicators by which to predict, with acceptable accuracy, the occurrence of DLE following rigid laryngoscopy. STUDY DESIGN: Fifty-six patients, each of whom had undergone a rigid laryngoscope procedure, were classified as belonging to either the DLE group (n = 19) or the control group (n = 37) for purposes of this prospective study. METHODS: All patients were given a physical examination, which encompassed the following nine measures: age, sex, modified Mallampati index (MMI), body mass index (BMI), hyoid-mental distance (HMD), thyroid-mental distance (TMD), thyroid mandible angle (TMA), horizontal thyroid distance (HTD), and vertical thyroid distance (VTD). Stepwise regression was employed on patient data to identify those with DLE. RESULTS The ages of patients in the DLE group ranged from 35 to 79 years, with a mean of 51.3 years. Among the nine variables, we found sex (P =.045, odds ratio = 69.159) and TMA (P =.004, odds ratio = 1.510) to be "reliable" DLE predictors. Using these two variables, 94.6% of study case patients could have been correctly classified preoperatively. Based on our comparison of case results, we found that a TMA value greater than 120 degrees in men and 130 degrees in women indicates a strong likelihood of DLE. CONCLUSIONS TMA is a sensitive, reliable, and useful predictor of DLE in both men and women. The combination of sex and TMA provide important initial clinical indicators that can alert a clinician regarding DLE probability. PMID- 14755219 TI - Microbiology of stents in laryngotracheal reconstruction. AB - OBJECTIVES: Granulation tissue often forms around a laryngotracheal stent, tracheostomy tube, or other airway prosthesis, especially if infection occurs. We studied the types and frequency of organisms colonizing stents used in pediatric laryngotracheal reconstruction. STUDY DESIGN: This prospective study included 21 patients undergoing 23 consecutive laryngotracheal reconstructions with stents between 1991 and 1999. METHODS: After endoscopic removal, each laryngotracheal stent was placed immediately in a sterile container and transported to the laboratory. Specimens for culture were obtained from biofilms on the stents and plated on agars for growth of aerobic, anaerobic, and fungal organisms. Culture results were analyzed with regard to patient age, duration of stenting, and graft type. RESULTS: All stents were colonized with more than one pathogen (range 2-7). The most frequent aerobic isolates were Streptococcus viridians, Pseudomonas aeruginosa, Staphylococcus aureus,Haemophilus influenza, and Neisseria species. Anaerobic organisms were isolated in 26% of cases. Candida species were isolated in 57% of the cases; patients whose stents were colonized with Candida were significantly (P =.007) older (mean 77.5 months) than those not colonized with this organism (mean 26.1 months) CONCLUSIONS: The antibiotic agents currently used for children undergoing laryngotracheal reconstruction target mainly aerobic organisms. Despite prophylactic measures, the incidence of granulation tissue formation is clinically significant, and the prevalence of anaerobic, including fungal, pathogens is high. Antibiotic therapy directed toward controlling anaerobic and fungal organisms could help in controlling local inflammation and thus granulation tissue formation. PMID- 14755220 TI - Effects of insulin-like growth factor-1 gene transfer on myosin heavy chains in denervated rat laryngeal muscle. AB - OBJECTIVES/HYPOTHESIS: To determine whether the myotrophic activity of human insulin-like growth factor (hIGF)-1 promotes restoration of normal myosin heavy chain (MHC) composition after nerve injury, MHC composition was analyzed after hIGF-1 gene transfer in denervated rat laryngeal muscle. STUDY DESIGN: Animal model to study effects of gene transfer on laryngeal paralysis. METHODS: In anesthetized rats, the left recurrent and superior laryngeal nerves are cut and suture ligated. A midline thyrotomy is performed, and the thyroarytenoid muscle is injected with a polyvinyl-based formulation containing a muscle specific expression system and hIGF-1 DNA (treatment group) or saline (control group). After 30 days, animals were killed, and the thyroarytenoid muscle was removed and processed for sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS PAGE). Densitometric measurements were obtained to determine composition of MHCs. RESULTS: As previously described, MHC composition in denervated laryngeal muscle was characterized by a decrease in type IIB and IIL and up-regulation of IIA/IIX. Compared with controls, hIGF-1 treated animals demonstrated a significant increase in expression of type IIB and IIL and a significant decrease in expression of type IIA/X. CONCLUSIONS: These findings suggest that the myotrophic effect of hIGF-1 gene transfer results in normalization of MHC composition in denervated muscle, with suppression of type IIA/X MHC and promotion of type IIL expression. PMID- 14755217 TI - Multicenter initiative seeking critical genes in respiratory papillomatosis. AB - OBJECTIVES: To determine the host genes that govern susceptibility to recurrent respiratory papillomatosis (RRP). RRP is caused by human papillomavirus (HPV) 6 and 11. Millions of babies are exposed during the birthing process, but relatively few develop the disease and the aggressiveness of the course is highly variable. Genetically encoded host susceptibility is postulated. Determining the host genes that govern susceptibility will enhance our understanding not only of RRP but also of host-viral interaction in general. STUDY DESIGN: A genome-wide association study on familial triads consisting of an RRP-affected child and his or her parents. Using the HapMap data from the human genome project, we will identify those alleles that are over-transmitted by the parents to their affected offspring as compared to those alleles that are under-transmitted. METHODS: Approximately 400 patients and their parents will be recruited through a collaboration between the Center for Genomic Sciences and the RRP Task Force. DNA will be extracted from blood specimens and viral typing will be performed on biopsy specimens. Patients will be genotyped using single nucleotide polymorphism (SNP) markers and compared to their respective parents' genotype using the transmission disequilibrium test. Both a genome scan and a candidate gene approach will be utilized. RESULTS Institutional Review Board authorization has been obtained at three hospitals and the process is underway at 18 more. Patient and parent recruitment has begun. Specimens have been forwarded to Pittsburgh, Pennsylvania, where the DNA has been extracted and is being stored. CONCLUSIONS: A novel approach combining a nationwide patient resource and the mapping power of the sub-centimorgan human haplotype map has been developed to elucidate the biological mechanisms of RRP by determining the genetically encoded susceptibilities of host-virus interaction. PMID- 14755221 TI - Inspiratory activation of the vocal cord adductor, part I: human study in patients with restricted abduction of the vocal cords. AB - OBJECTIVES/HYPOTHESIS: In patients with restricted abduction of the vocal cords, it has generally been accepted that glottis narrowing with laryngeal stridor during inspiration is attributed to static and passive obstruction of the glottis. However, active glottis narrowing can also be contributory. We tested the hypothesis that the vocal cord adductor is activated during inspiration in patients with restricted abduction of the vocal cords. STUDY DESIGN: Electromyographic evaluation of vocal cord adductor activity in patients with restricted abduction of the vocal cords. METHODS: Five patients with restricted abduction of the vocal cords who had stridor with mild to severe dyspnea during wakefulness were anesthetized with propofol. We recorded the adductor muscle electromyogram during breathing through a laryngeal mask airway while observing the vocal cord movement endoscopically. In three patients who had undergone tracheostomy, we also recorded adductor firing patterns not only while closing but also while opening the tracheostoma. RESULTS: The adductor was activated during inspiration, and the glottis was narrowed in accordance with inspiratory stridor. This adductor inspiratory activity was abolished by opening the tracheostoma in the tracheostomized patients. CONCLUSION: Not only static or passive glottis narrowing but also active narrowing may contribute to inspiratory flow limitation in patients with restricted abduction of the vocal cords. This active glottis narrowing is probably induced by an airway reflex. PMID- 14755223 TI - Parotidectomy using the Harmonic scalpel. PMID- 14755222 TI - Inspiratory activation of the vocal cord adductor, part II: Animal study in the cat. AB - OBJECTIVES/HYPOTHESIS: The authors have shown previously that the vocal cord adductor is activated during inspiration in patients with vocal cord abduction impairment and that this adductor inspiratory activity is abolished by relief from inspiratory tracheal negative pressure by opening the tracheostoma. (Shiba K. Isono S, Sekita Y, Tanaka A. Inspiratory activation of the vocal cord adductor, Part I: human study in patients with restricted abduction of the vocal cords. Laryngoscope 2004;114:372-375). The authors hypothesized that insufficient opening of the glottis during inspiration generates strong negative pressure in the trachea and that this negative pressure triggers an airway reflex that activates the adductor. STUDY DESIGN: Experimental study of the mechanism of laryngeal obstruction using an animal model of restricted abduction of the vocal cords. METHODS: To identify such an airway reflex, the authors recorded the adductor electromyogram in anesthetized cats whose vocal cords were mechanically adducted by stitching both cords together. To determine whether this reflex modulation of adductor activity is induced through afferents from the larynx or from the lower airway, the authors applied negative pressure to the subglottic space and lower airway separately. RESULTS: The adductor was activated during inspiration with powerful negative pressure in the trachea. Negative pressure in the subglottic space had a more marked effect on the adductor activity than did pressure in the lower airway. The adductor inspiratory activity was virtually abolished by laryngeal deafferentation. CONCLUSION: Glottal narrowing during inspiration reflexly activates the vocal cord adductor. This paradoxical inspiratory-related adductor activation is induced by an airway reflex triggered mainly through afferents from the larynx and probably contributes to stridor and dyspnea in patients with laryngeal obstruction. PMID- 14755224 TI - Direct tissue expansion of the contracted nose. PMID- 14755225 TI - Permeatal temporalis fascia graft harvesting for minimally invasive myringoplasty. PMID- 14755226 TI - Canadian report: ICP needs across the health care continuum. PMID- 14755227 TI - Development of a resource model for infection prevention and control programs in acute, long term, and home care settings: conference proceedings of the Infection Prevention and Control Alliance. AB - There is mounting concern about the impact of health care restructuring on the provision of infection prevention services across the health care continuum. In response to this, Health Canada hosted two meetings of Canadian infection control experts to develop a model upon which the resources required to support an effective, integrated infection prevention and control program across the health care continuum could be based. The final models project the IPCP needs as three full time equivalent infection control professionals/500 beds in acute care hospitals and one full time equivalent infection control professional/150-250 beds in long term care facilities. Non human resource requirements are also described for acute, long term, community, and home care settings. PMID- 14755228 TI - Evaluation of a public health Salmonella surveillance system in King County, Washington. AB - OBJECTIVES: Salmonella infection is the most common cause of foodborne bacterial outbreaks and deaths in the United States. The effectiveness of Salmonella surveillance for detecting outbreaks depends on timeliness of reporting. We evaluated the public health Salmonella surveillance system in King County, Washington, during an outbreak and at baseline. METHODS: We assessed the timeliness of the Salmonella surveillance system in King County using the Public Health-Seattle & King County Laboratory (County PHL) database from 1998 to 1999. We determined median days for key steps involved in the Salmonella identification and reporting process and the percentage of suspected Salmonella isolates confirmed. Time intervals for key steps during a Salmonella outbreak were compared to baseline surveillance. RESULTS: Of the 652 suspected Salmonella isolates sent to the County PHL from 22 clinical laboratories, 617 (94.6%) were confirmed as Salmonella. Salmonella confirmation rates improved from 1998 to 1999, and 41% of the submitting laboratories, contributing 32.4% of the isolates, had 100% confirmation rates for both years. The median total identification time during the outbreak did not differ significantly from baseline (13 days vs 17 days). The time interval for serotyping contributed most to the total identification time. CONCLUSIONS: King County's Salmonella surveillance system requires more than 2 weeks to confirm and report serotype results for Salmonella isolates. Variation in total identification time depends on serotyping time. A more detailed study of other surveillance systems may identify approaches to decrease total identification time. PMID- 14755229 TI - Bacterial contamination of multiple-dose vials: a prevalence study. AB - BACKGROUND: Two patients died of a meningitis caused by Pseudomonas aeruginosa in a hospital in Germany in July 2001, their infections having been caused by a contaminated contrast media (iomeprol [Imeron]) used as a multiple-dose vial (MDV) over 8 days. Therefore, a prevalence study was performed to investigate the use and contamination of multiple-use vials in a tertiary hospital. METHODS: In a 1300-bed hospital on a specific day in November 2001, all used MDVs were collected by the infection control nurses. Information was recorded about the medication, labeling of vials, storing temperature, wards, and dates of opening. Each vial was also tested for sterility. RESULTS: Opened vials were to be found in all wards. Of the 227 vials available, 1 vial and 1 spike were contaminated with Staphylococcus epidermidis (contamination rate 0.9%; 95% CI, 0.3-2.1). The opening dates were marked on only 114 (50%) MDVs, 15 (13%) of which had already expired. Only 44 (19%) MDVs had been stored in the refrigerator, whereas 109 MDVs contained medications without any preserving agent. CONCLUSION: Results revealed somewhat risky handling of MDVs. In light of a possible high risk in this hospital of about 1 contaminated MDV per day, and in view of many reported outbreaks induced by contaminated MDVs, the following infection control measures were encouraged: alcohol hand hygiene, the disinfection of gums, observance of the manufacturer's recommendations, appropriate storing temperatures, marking the opening time, and avoiding the multiple use of medications not containing preserving agents. PMID- 14755230 TI - Polymerase chain reaction used for the detection of airborne Mycobacterium tuberculosis in health care settings. AB - BACKGROUND: Tuberculosis (TB) has re-emerged as a major infectious disease in several areas in the world. Airborne tubercle bacilli produced by individuals with pulmonary TB and droplet nuclei remain suspended in the air for a long time. This study attempts to use a sensitive polymerase chain reaction (PCR) analytic method coupled with a filter sampling method to detect the presence of airborne Mycobacterium tuberculosis (MTb) in health care settings. METHODS: Patients with TB history were recruited from a medical intensive care department and negative pressure isolation rooms at Chang Gung Memorial Hospital in Taichung City, Taiwan, Republic of China. The exhaled gas from different patients with TB who were mechanically ventilated in the intensive care department was collected using a polycarbonate (PC) membrane or polytetrafluoroethylene (PTFE) filter. Airborne MTb concentrations in air exhausted through a bacterial filter attached to a mechanic ventilator were studied. Indoor air samples were taken through a 3-piece cassette with a filter (PC/PTFE) in patient rooms. MTb concentrations in these filters were analyzed using the PCR method. RESULTS: Overall, 75% (12/16) of the exhaled-gas samples in PTFE filters and 25% (4/16) of samples in PC filters were detected as having positive MTb-specific DNA products. Exhausted air from the bacterial filters in mechanic ventilators was found to have positive PCR results (57% in PC filters and 14% in PTFE filters) for MTb. Both negative-pressure isolation rooms and outpatient department areas in the TB center had positive samples (40%-60% in PC/PTFE filters) containing MTb by PCR amplification. CONCLUSIONS: Health care settings were sufficient for various risk factors, including physical, chemical, and biologic hazards. Infectious agents are produced by aerosolization that results in human respiratory-related diseases. The indoor air quality of the entire hospital environment should, therefore, be monitored by health care personnel and the public. PMID- 14755231 TI - Incidence and outcome of nosocomial infections in female burn patients in Shiraz, Iran. AB - BACKGROUND: Burn patients are at risk for acquiring infection because of their destroyed skin barrier and suppressed immune system, compounded by prolonged hospitalization and invasive therapeutic and diagnostic procedures. Most studies on infection in burn patients focus on burn wound infection, whereas other nosocomial infections in this patient group are not described well. OBJECTIVE: This study attempts to assess the incidence of nosocomial infection in the Ghotbeddin burn center of Shiraz. METHODS: The study was conducted prospectively during a period of 11 months from December 2000 to November 2001. All patients presenting with no signs and symptoms of infection within the first 48 hours of admission were included and examined for detecting 4 types of nosocomial infection: burn infection, urinary tract infection, pneumonia, and bloodstream infection. Centers for Disease Control and Prevention National Nosocomial Infection Surveillance system criteria were applied. RESULTS: One hundred six female patients met the inclusion criteria. Ninety-one (85.8%) acquired at least 1 infection (44.7 per 1000 patient-days), including 91 with burn infection, 28 with urinary tract infection, 56 with pneumonia, and 30 with bloodstream infection, which gives 446.7, 137.5, and 275, and 147.3 infections per 1000 patient-days, respectively. CONCLUSIONS: Considering the high incidence of nosocomial infections in our center, implementation of improved infection control practices and policies is required, and a comprehensive education campaign for all health care workers is urgently needed. PMID- 14755232 TI - Distinctive bacteria-binding property of cloth materials. AB - BACKGROUND: Nosocomial infections may be caused by pathogens that are transmitted from the hands or clothes of hospital personnel. Handwashing has been evaluated as effective against the spread of pathogens, but transmission through clothes has been little investigated. Evaluation of bacterial adherence to clothes is difficult because of the nonuniform amount of water absorbance by cloth. Therefore, we measured binding of bacteria to cloth fibers made of cotton, nylon, polyester, acrylic, or sheep's wool and tried to characterize bacterial binding to cloth. METHODS: We chose to study the opportunistic pathogens Staphylococcus aureus and Pseudomonas aeruginosa. Cloth fibers were incubated with bacterial suspensions in silicone-coated tubes. We evaluated the reduction of numbers of bacteria in solutions incubated with the fibers and calculated binding ratios of bacteria to the fibers. RESULTS: Polyester or acrylic fibers bound S aureus and P aeruginosa at high ratios (>80%), but cotton fibers bound them at low ratios (<10%). Nylon fibers bound S aureus at low ratios, but P aeruginosa at intermediate ratios. CONCLUSION: The results suggested that polyester, acrylic, or wool clothes could be good carriers of S aureus and P aeruginosa and thus should be covered with cotton clothes to minimize the spread of the pathogens. PMID- 14755233 TI - Rapid assessment of injection practices in Mongolia. AB - BACKGROUND: Anecdotal reports suggest that unsafe injections may transmit blood borne pathogens in Mongolia. METHODS: The Ministry of Health of Mongolia collected information on injection practices, their determinants, and their consequences through interviews and observations of a small convenience sample of prescribers, injection providers, and members of the general population. RESULTS: The 65 members of the general population reported receiving an average of 13 injections per year. New, single-use injection devices were used in the 20 health care facilities visited. There were breaks in infection control practices while administering injections, including observations of 500-mL intravenous infusion bottles used as multiple-dose diluent vials and 8 of the 28 providers (28%) reporting reuse of device on the same patient. Injection providers reported 2.6 needle-stick injuries per year. Contaminated sharps were burned in a drum. Among persons interviewed, 19 of the 21 prescribers (90%) and 49% of the population were aware of the potential risk of HIV transmission through unsafe injections. CONCLUSIONS: A multidisciplinary initiative is necessary to achieve safe and appropriate use of injections in Mongolia through (1) behavior change, (2) increasing availability of injection devices and sharps boxes, and (3) appropriate sharps waste management. PMID- 14755234 TI - Contact investigation of a case of active tuberculosis. AB - BACKGROUND: The Veterans Administration (VA), Marion, Illinois, following Public Health and Veterans Affairs guidance has an excellent tuberculosis (TB) control program with all employees included. METHODS: This VA is a low-risk facility for TB, according to the Centers for Disease Control and Prevention guidelines. A patient presented with active TB in January 2002. Follow-up was complicated and enlarged to the community, adjacent state, and notification of Centers for Disease Control and Prevention. The concentric circle approach in identifying highest-risk contacts had to be modified and expanded because of the immunocompromised condition of some of the contacts. RESULTS: After identification of active TB, contacts were identified. Initially 108 employees and 42 patients were reviewed for follow-up at the VA, with 98 employees and 27 patients actually tested. At 3-month follow-up there were 2 conversions. CONCLUSION: Because of excellent participation in the employee tuberculin skin testing program, conversion was readily identified among staff. Contact investigation of a case of TB requires both diligence in the program and prompt, effective communication and action between state and local health departments and hospitals. PMID- 14755235 TI - Rapid control of an outbreak of Staphylococcus aureus on a neonatal intensive care department using standard infection control practices and nasal mupirocin. AB - BACKGROUND: Staphylococcus aureus is a common pathogen in neonatal intensive care departments, causing significant morbidity, mortality, and cost. Frequently, S aureus outbreaks may last for months or years. After a cluster of 4 clinically significant S aureus infections in a 7-day period in our 35-bed neonatal intensive care department, we immediately introduced standard outbreak control measures. Unique to our approach was the addition of immediate nasal mupirocin treatment of all staff members and selected patients. METHODS: Patients were screened for S aureus colonization and were cohorted with separate caregivers. S aureus isolates were submitted to a reference laboratory for pulse-field gel electrophoretic typing. Infection control practices were emphasized and education was provided for staff, physicians, and parents of patients. All caregivers and selected patients were treated immediately with nasal mupirocin. Cohorting was maintained until all patients who were colonized or infected were discharged. RESULTS: A total of 5 patients were found to be infected and 4 of 19 patients tested were found to be colonized during the study period. Patients who were infected were successfully treated. Secondary colonization and infection did not occur after implementation of controls. CONCLUSIONS: Rapid and comprehensive implementation of standard outbreak controls along with immediate treatment of direct care staff and patients with nasal mupirocin successfully controlled this outbreak within 4 weeks and no further cases have been noted. PMID- 14755236 TI - A tool to assess barriers to adherence to hand hygiene guideline. AB - This Brief Report presents results of preliminary testing of a tool, "Attitudes Regarding Practice Guidelines," adapted from previous work by Cabana et al to examine barriers to adherence to practice guidelines. In a small sample of 21 clinicians (10 infectious disease physicians and 11 clinical nurses), the test retest reliability was.86 and Chronbach alpha was.80, indicating acceptable levels of stability. It is recommended that the instrument undergo further testing and adaptation as a measure of potential barriers to adherence to practice guidelines in general, and to the new Hand Hygiene Guideline for Healthcare Settings specifically. PMID- 14755237 TI - Stress ulcer prophylaxis and risk of developing ventilator-associated pneumonia. PMID- 14755238 TI - Identification of genes whose expression is upregulated in lung adenocarcinoma cells in comparison with type II alveolar cells and bronchiolar epithelial cells in vivo. AB - To identify genes whose expression is upregulated in lung adenocarcinoma (AdC) cells in comparison with noncancerous peripheral lung epithelial cells, type II alveolar cells and bronchiolar epithelial cells, as well as AdC cells, were isolated by laser capture microdissection, and subjected to cDNA microarray analysis of 637 human cancer-related genes. Each of the component cells was obtained from several different individuals and analysed independently. As a comparison, two lung AdC cell lines and two primarily cultured normal lung epithelial cell lines were also subjected to cDNA microarray analysis. Four genes, TOP2A, MMP15, MX2 and KOC1, were commonly upregulated in microdissected AdC cells in comparison with microdissected epithelial cells. Hierarchical clustering analysis revealed that differences in gene-expression profiles were more evident between cultured and uncultured cells than between cancerous and noncancerous cells. To further identify the common molecular targets of AdC cells in vivo, quantitative real-time RT-PCR was performed against the four genes upregulated by cDNA microarray analysis. The TOP2A, MMP15, MX2 and KOC1 genes were overexpressed in 10/10 (100%), 8/10 (80%), 5/10 (50%) and 3/10 (30%) microdissected AdC cell samples, respectively, in comparison with any of nine independently microdissected noncancerous epithelial cell samples. The TOP2A gene was commonly overexpressed in lung AdC cells, as previously reported. In addition, the MMP15 and MX2 genes were identified, for the first time, as being commonly overexpressed in lung AdC cells. These results strongly indicate that the MMP15 and MX2 genes could be novel markers for molecular diagnosis and therapy of lung AdC. PMID- 14755239 TI - Carcinogen-specific targeting of chromosome 12 for loss of heterozygosity in mouse lung adenocarcinomas: implications for chromosome instability and tumor progression. AB - Genotoxic carcinogens exert their tumorigenic effects in part by inducing genomic instability. We recently showed that loss of heterozygosity (LOH) on chromosome 12 associates significantly with the induction of chromosome instability (CIN) by the likely human lung carcinogen 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) and vinyl carbamate (VC) during mouse lung carcinogenesis. Here, we demonstrate the carcinogen specificity of this event and its effect on lung tumor evolution. LOH on chromosome 12 was observed in 45% of NNK-induced, 59% of VC induced, 58% of aflatoxin B1 (AFB1)-induced, 14% of N-ethyl-N-nitrosourea (ENU) induced and 12% of spontaneous lung adenocarcinomas. The frequency of LOH in each of the carcinogen-induced groups, except ENU, was significantly higher than in the spontaneous group (P<0.001). Deletion mapping revealed four potential candidate regions of 1-4 centiMorgans suspected to contain targeted tumor suppressor genes, with at least one expected to have a role in CIN. The relationship between LOH on chromosome 12 and additional chromosomal alterations occurring during lung tumor progression was also examined. LOH on chromosomes 1 and 14 were moderately frequent during malignant progression in tumors from all treatment groups, occurring in 21-35 and 18-33% of tumors. However, these alterations showed significant concurrence with LOH on chromosome 12 in VC-, NNK- and AFB1-induced tumors (P<0.05). The results suggest that a carcinogen-selective mechanism of lung cancer induction involves the frequent inactivation of genes on chromosome 12, including a stability gene that evidently promotes the evolutionary selection of additional chromosomal alterations during malignant progression. PMID- 14755240 TI - Expression of alpha-catenin in alpha-catenin-deficient cells results in a reduced proliferation in three-dimensional multicellular spheroids but not in two dimensional monolayer cultures. AB - alpha-Catenin is an intracellular protein that associates with the carboxy terminal region of cadherin, a cell adhesion molecule, via beta-catenin or gamma catenin (plakoglobin). Linkage of cadherin to the cytoskeleton by catenins is required for full cadherin activity. Following transfection of an alpha-catenin deficient colon carcinoma cell line with a series of alpha-catenin constructs, we discovered that the restoration of alpha-catenin expression results in reduced proliferation in three-dimensional multicellular spheroids, but not in two dimensional monolayer cultures. The cellular function of alpha-catenin has not been compared between cells in three- and two-dimensional culture; this is the first evidence that growth regulation in three-dimensional cultures requires signaling mediated by alpha-catenin. Two classes of constructs, containing deletions in either the central segment or the COOH terminus of the molecule, both induced morphological changes, including cell compaction, and suppressed cell growth in three-dimensional cultures. In alpha-catenin-expressing cells, inhibition of cadherin cell adhesion by treatment with anti-E-cadherin antibodies resulted in a similar phenotype as that observed following the loss of alpha catenin. Therefore, both the homophilic interaction of the cadherin extracellular domain and the linkage of the cadherin cytoplasmic domain to the actin cytoskeleton by alpha-catenin are necessary for growth control in three dimensional culture. PMID- 14755241 TI - Gene-expression profiling reveals distinct expression patterns for Classic versus Variant Merkel cell phenotypes and new classifier genes to distinguish Merkel cell from small-cell lung carcinoma. AB - Merkel cell carcinoma (MCC) is a rare aggressive skin tumor which shares histopathological and genetic features with small-cell lung carcinoma (SCLC), both are of neuroendocrine origin. Comparable to SCLC, MCC cell lines are classified into two different biochemical subgroups designated as 'Classic' and 'Variant'. With the aim to identify typical gene-expression signatures associated with these phenotypically different MCC cell lines subgroups and to search for differentially expressed genes between MCC and SCLC, we used cDNA arrays to profile 10 MCC cell lines and four SCLC cell lines. Using significance analysis of microarrays, we defined a set of 76 differentially expressed genes that allowed unequivocal identification of Classic and Variant MCC subgroups. We assume that the differential expression levels of some of these genes reflect, analogous to SCLC, the different biological and clinical properties of Classic and Variant MCC phenotypes. Therefore, they may serve as useful prognostic markers and potential targets for the development of new therapeutic interventions specific for each subgroup. Moreover, our analysis identified 17 powerful classifier genes capable of discriminating MCC from SCLC. Real-time quantitative RT-PCR analysis of these genes on 26 additional MCC and SCLC samples confirmed their diagnostic classification potential, opening opportunities for new investigations into these aggressive cancers. PMID- 14755242 TI - HIV-1 Tat increases cell survival in response to cisplatin by stimulating Rad51 gene expression. AB - Tat is an early regulatory protein of human immunodeficiency virus type 1, which plays a central role in the pathogenesis of AIDS by stimulating transcription of the viral genome and impairing several important cellular pathways during the progression of the disease. Here, we investigated the effect of Tat on cell response to DNA damage. Our results indicate that Tat production causes a noticeable increase in the survival rate of PC12 cells upon their treatment with genotoxic agents. Single-cell gel electrophoresis studies revealed reduced DNA breakage in PC12-Tat cells upon cisplatin treatment relative to the control cells. Furthermore, cytogenetic data exhibited less chromosomal damage in Tat producing cells after recovery from cisplatin treatment, corroborating electrophoretic data. Examination of several proteins involved in the control of DNA repair showed elevated levels of Rad51, a key regulator of homologous recombination in cells expressing Tat. On the other hand, the level of Ku70, one of the components of the nonhomologous end-joining repair pathway, was slightly decreased in cells expressing Tat. Using a fluorescence-based assay, we demonstrated that repair of DNA double-strand breaks via homologous recombination is increased in Tat-producing cells. The results from in vitro nonhomologous end joining assay revealed a reduced ability of protein extract from PC12-Tat cells compared to PC12 cells in rejoining linearized DNA. These observations ascribe a new role for Tat in host genomic integrity, perhaps by affecting the expression of genes involved in DNA repair. PMID- 14755243 TI - beta-Catenin and TGFbeta signalling cooperate to maintain a mesenchymal phenotype after FosER-induced epithelial to mesenchymal transition. AB - Several signalling pathways contribute to the regulation of epithelial to mesenchymal transition (EMT), either during developmentally regulated processes or in cancer progression and metastasis. Induction of EMT in fully polarized mouse mammary epithelial cells (EpH4) by an inducible c-fos estrogen receptor (FosER) oncoprotein involves loss of E-cadherin expression, nuclear translocation of beta-catenin, and autocrine production of TGFbeta. Reporter assays demonstrate that both beta-catenin/LEF-TCF- and TGFbeta-Smad-dependent signalling activities are upregulated, probably coregulating mesenchymal-specific gene expression during EMT. Stable expression of E-cadherin in mesenchymal FosER cells decreased beta-catenin activity and reduced cell proliferation. However, these cells still exhibited a defect in epithelial polarization and expressed E-cadherin/beta catenin complexes in the entire plasma membrane. On the other hand, inhibition of TGFbeta-Smad signalling in mesenchymal FosER cells induced flat, cobblestone-like clusters of cells, which relocalized beta-catenin to the plasma membrane but still lacked detectable E-cadherin. Interestingly, inhibition of TGFbeta signalling in the E-cadherin-expressing mesenchymal FosER cells caused their reversion to a polarized epithelial phenotype, in which E-cadherin, beta-catenin, and ZO-1 were localized at their correct lateral plasma membrane domains. These results demonstrate that loss of E-cadherin can contribute to increased LEF/TCF beta-catenin signalling, which in turn cooperates with autocrine TGFbeta signalling to maintain an undifferentiated mesenchymal phenotype. PMID- 14755244 TI - The c-Rel transcription factor and B-cell proliferation: a deal with the devil. AB - Activation of the Rel/NF-kappaB signal transduction pathway has been associated with a variety of animal and human malignancies. However, among the Rel/NF-kappaB family members, only c-Rel has been consistently shown to be able to malignantly transform cells in culture. In addition, c-rel has been activated by a retroviral promoter insertion in an avian B-cell lymphoma, and amplifications of REL (human c-rel) are frequently seen in Hodgkin's lymphomas and diffuse large B-cell lymphomas, and in some follicular and mediastinal B-cell lymphomas. Phenotypic analysis of c-rel knockout mice demonstrates that c-Rel has a normal role in B cell proliferation and survival; moreover, c-Rel nuclear activity is required for B-cell development. Few mammalian model systems are available to study the role of c-Rel in oncogenesis, and it is still not clear what features of c-Rel endow it with its unique oncogenic activity among the Rel/NF-kappaB family. In any event, REL may provide an appropriate therapeutic target for certain human lymphoid cell malignancies. PMID- 14755245 TI - Modulation of different clusterin isoforms in human colon tumorigenesis. AB - Clusterin is a ubiquitous secretory heterodimeric disulfide-linked glycoprotein, which is implicated in several physiological processes, including immune regulation, cell adhesion and morphological transformation, lipid transportation, tissue remodelling, membrane recycling and cell-cell interactions. A large number of studies have focused their interest on clusterin gene products as mediators of cell cycle progression and cell death induction, although data on the different isoforms and their role in the different cell processes are still obscure. Recently, an increased clusterin expression in breast cancer has been reported. In order to elucidate the role of clusterin in tumor progression and whether one of its isoforms is preferentially expressed in tumorigenesis, we examined its presence throughout the different steps of human colon carcinoma, one of the best characterized models of human tumor progression. The immunohistochemical observation of 30 bioptic and surgical colon specimens demonstrated a cell compartment clusterin translocation from the nucleus to the cytoplasm directly related to tumor progression. In fact, a nuclear localization found in healthy colonic mucosa is consistent with the involvement of the proapoptotic nuclear form in the regulation of cell cycle progression and in cell death induction. The progression towards high-grade and metastatic carcinoma leads to cytoplasmic clusterin distribution. Protein extracts from freshly isolated cells of the same patients confirm in high-grade carcinomas with metastatic nodes the complete loss of the proapoptotic nuclear form and a cytoplasmic overexpression of the highly glycosylated form. Data obtained from in vitro experiments confirm that this form is released in the extracellular space and corresponded to the fully glycosylated one. These data suggest that the controversial data on clusterin function in tumors may be related to the pattern shift of its isoform production. As the secreted form of clusterin is correlated to cell matrix formation, cell membrane remodeling and cell-cell adhesion, the overexpression of this form in highly aggressive tumors and metastatic nodes could be a potential new prognostic and predictive marker for colon carcinoma aggressiveness. PMID- 14755246 TI - Generation and analysis of melanoma SAGE libraries: SAGE advice on the melanoma transcriptome. AB - In this study, we generated three SAGE libraries from melanoma tissues. Using bioinformatics tools usually applied to microarray data, we identified several genes, including novel transcripts, which are preferentially expressed in melanoma. SAGE results converged with previous microarray analysis on the importance of intracellular calcium and G-protein signaling, and the Wnt/Frizzled family. We also examined the expression of CD74, which was specifically, albeit not abundantly, expressed in the melanoma libraries using a melanoma progression tissue microarray, and demonstrate that this protein is expressed by melanoma cells but not by benign melanocytes. Many genes involved in intracellular calcium and G-protein signaling were highly expressed in melanoma, results we had observed earlier from microarray studies (Bittner et al., 2000). One of the genes most highly expressed in our melanoma SAGE libraries was a calcium-regulated gene, calpain 3 (p94). Immunohistochemical analysis demonstrated that calpain 3 moves from the nuclei of non-neoplastic cells to the cytoplasm of malignant cells, suggesting activation of this intracellular proteinase. Our SAGE results and the clinical validation data demonstrate how SAGE profiles can highlight specific links between signaling pathways as well as associations with tumor progression. This may provide insights into new genes that may be useful for the diagnosis and therapy of melanoma. PMID- 14755248 TI - Enhancement of p53 sequence-specific binding by DNA supercoiling. AB - Using a new competition assay, we investigated the effect of DNA negative supercoiling on the DNA sequence-specific binding (SSDB) of human wild-type (wt) p53 protein. We found that supercoiled (sc) pBluescript DNAs with different inserted p53 target sequences were stronger competitors than a mixture of scDNA pBluescript with the given 20-mer target oligodeoxynucleotide. ScDNAs were always better competitors than their linearized or relaxed forms. Two DNAs with extruded cruciforms within the target sequence were the best competitors; removal of the cruciforms resulted in a decrease of competitor strength. In contrast to the full length wt p53, the deletion mutant p53CDelta30 and the p53 core domain (93-312 aa) showed no enhancement of p53 SSDB to scDNA, suggesting that, in addition to the p53 core domain, the C-terminal was involved in this binding. We conclude that cruciforms and DNA bends contribute to the enhancement of p53 SSDB to scDNA and that the DNA supercoiling is an important determinant in the p53 sequence specific binding. Supercoiling may thus play a significant role in the complex p53-regulatory network. PMID- 14755247 TI - Attenuation of the p53 response to DNA damage by high cell density. AB - The p53 tumor suppressor is critical for preventing cancer progression. Numerous observations suggest that p53 function can be modulated by the cells' microenvironment. We addressed specifically the impact of cell crowding on the induction of p53 by DNA damage. We report that cell crowding attenuates markedly p53 upregulation, transcriptional activation and subsequent p53-dependent apoptosis following exposure to genotoxic stress. The p53 protein remains short lived in confluent cultures regardless of the extent of DNA damage, even though it undergoes efficient phosphorylation on the mouse equivalent of human p53 serine 15. This inhibitory effect of cell crowding is not a secondary consequence of density-dependent cell cycle arrest (contact inhibition). Microscopic examination indicates that dense cultures display prominent cadherin-mediated cell-cell junctions, and only poor cell-matrix focal adhesions, whereas sparse cells possess conspicuous matrix adhesions and essentially no cell-cell contacts. High-density cell culture might recapitulate the microenvironment of cells in a living organism, where the response of p53 to DNA damage is reported to be low in some organs and ages. The impact of cell density on p53 activation may have important bearings on the involvement of p53 in tumor suppression and the cellular response to anticancer therapy. PMID- 14755249 TI - p53-dependent and p53-independent pathways for radiation-induced immature thymocyte differentiation. AB - The pre-T-cell receptor (TCR) delivers essential survival/differentiation signals to the developing thymocytes. Severe combined immunodeficient (SCID) and recombination-activating gene (RAG)-deficient mice are unable to assemble antigen receptor genes, and therefore cannot express a pre-TCR. Consequently, T lymphocyte differentiation is arrested at an early stage in the thymus of these animals, and immature thymocytes are eliminated through apoptotic processes. This maturation arrest can be relieved and thymocyte differentiation rescued after the exposure of these mice to whole-body gamma-irradiation. Whereas the promotion of immature thymocyte survival/differentiation was shown to require p53 activity in irradiated SCID mice, it was suggested, on the other hand, that p53 activation prevents immature thymocytes survival/differentiation in irradiated RAG-deficient mice. However, SCID mice have impaired responses to ionizing radiation. In this paper, we analysed p53 requirement in radiation-induced thymocyte differentiation in CD3epsilon(Delta5/Delta5) mice, where pre-TCR deficiency also results in an early block of lymphocyte development. Our results show at the cellular and molecular levels that, in this DNA repair-proficient model, irradiation-induced thymocyte differentiation proceeds either by a p53-dependent or by a p53 independent pathway, which differ in their sensitivity to the radiation dose delivered. PMID- 14755250 TI - An RNF11: Smurf2 complex mediates ubiquitination of the AMSH protein. AB - RING-finger proteins play crucial roles in ubiquitination events involved in diverse cellular processes including signal transduction, differentiation and apoptosis. Most of the RING-finger proteins have E3-ubiquitin ligase activity. RNF11 is a small RING-finger protein and harbors a RING-H2 domain and a PY motif that could facilitate protein:protein interaction(s) involved in oncogenesis. To isolate RNF11 protein partners and determine its role in normal and cancer cells, we performed yeast two-hybrid screening. Among 18 in-frame positive clones, three were found to be ZBRK1, Eps15 and AMSH (associated molecule with the SH3 domain of STAM). ZBRK1 is a KRAB domain containing Zinc-finger protein and is known to repress target gene transcription in a BRCA1-dependent manner. Eps15 is monoubiquitinated and is part of an essential complex involved in the endocytosis of plasma membrane receptors via the clathrin-mediated internalization pathway. Recent studies have shown that AMSH protein is involved in BMP/TGF-beta signaling pathway by binding to Smad6 and Smad7. The association of RNF11 with these binding partners suggests that it would be involved in biological processes such as gene transcription, BMP/TGF-beta signaling and ubiquitination-associated events. Previously, we have shown that RNF11 interacts with the HECT-type E3 ligases AIP4 and Smurf2. Here, we show that RNF11 binds to AMSH in mammalian cells and that this interaction is independent of the RNF11 RING-finger domain and the PY motif. Our results also demonstrate that AMSH is ubiquitinated by Smurf2 E3 ligase in the presence of RNF11 and that a consequent reduction in its steady-state level requires both RNF11 and Smurf2. RNF11 therefore recruits AMSH to Smurf2 for ubiquitination, leading to its degradation by the 26S proteasome. The potential functions of RNF11-mediated degradation of AMSH in breast cancer are discussed. PMID- 14755251 TI - The ATR-p53 pathway is suppressed in noncycling normal and malignant lymphocytes. AB - Chronic lymphocytic leukaemia (CLL) results from the accumulation of apoptosis resistant clonal B cells that are arrested in G0/G1, and is heterogeneous with respect to clinical outcome. An aggressive form of the disease is identified by an impaired p53 response to ionizing radiation (IR). This is associated with inactivating mutations of either p53 or ATM, a regulator of p53 activated by IR induced DNA damage. Since other forms of DNA damage activate p53 via ATR, a kinase closely related to ATM, abnormalities of the ATR-p53 pathway also have the potential to result in p53 dysfunction. We therefore tested cases of CLL for abnormal p53 responses to ultraviolet irradiation (UVC), a known activator of ATR, to screen for additional forms of p53 dysfunction. CLL cells and normal peripheral blood mononuclear cell (PBMC) preparations (predominantly noncycling lymphocytes) were treated with UVC and assessed for p53 responses. In all of the CLL cases and PBMC preparations tested, we were unable to detect p53 accumulation, phosphorylation or transcriptional consequences in response to UVC induced DNA damage. The most likely explanation for the absence of UVC-induced p53 activation in CLL and normal lymphocytes was that, in contrast to other cell types, the UVC-induced ATR pathway was inactive. This notion was confirmed by showing that ATR protein was absent or undetectable in all of the cases of CLL and normal PBMCs screened. This was an unexpected finding because ATR was thought to be essential for the viability of somatic cells and for normal human and murine embryonic development. An obvious difference between the cell lines used as positive controls for ATR antibodies and the CLL cells/PBMCs was that the former were actively cycling while the latter were quiescent. We therefore hypothesized that the ATR-p53 pathway is selectively downregulated in noncycling lymphocytes. To test this, we induced cycling in the T-cell fraction of PBMC preparations and demonstrated that ATR protein expression was restored. Furthermore, p53 was upregulated and phosphorylated in response to UVC in these cells. Our data support the conclusion that the ATR-p53 pathway is suppressed in noncycling lymphocytes via ATR downregulation. We tentatively suggest that this repressed DNA damage response may have evolved to protect quiescent lymphocytes from the potential for p53-dependent apoptosis in the face of some forms of endurable genotoxic stress. If this is the case, DNA repair and genome stability might be compromised in quiescent lymphocytes with potentially negative consequences. PMID- 14755252 TI - The helix-loop-helix protein ID1 localizes to centrosomes and rapidly induces abnormal centrosome numbers. AB - ID1 is a member of the inhibitor of DNA binding/differentiation (ID) family of dominant negative helix-loop-helix transcription factors. ID-proteins have been implicated in the control of differentiation and transcriptional modulation of various cell cycle regulators and high levels of ID1 expression are frequently detected in various cancer types. However, it is unclear whether ID1 is a marker of highly proliferative cancer cells or whether it directly contributes to the tumor phenotype. A detailed analysis of ID1-expressing human cells revealed that a fraction of ID1 localizes to centrosomes. Ectopic expression of ID1 in primary cells and tumor cell lines resulted in accumulation of cells with abnormal centrosome numbers. There was no evidence for centrosomal localization or induction of centrosome abnormalities by the other ID family members. Hence, ID1 may contribute to oncogenesis not only by inhibiting transcriptional activity of basic helix-loop-helix transcription factors and abrogate differentiation but also by subverting centrosome duplication. PMID- 14755253 TI - Inositol pentakisphosphate promotes apoptosis through the PI 3-K/Akt pathway. AB - Phosphoinositide 3-kinase (PI 3-K) is implicated in a wide array of biological and pathophysiological responses, including tumorigenesis, invasion and metastasis, therefore specific inhibitors of the kinase may prove useful in cancer therapy. We propose that specific inositol polyphosphates have the potential to antagonize the activation of PI 3-K pathways by competing with the binding of PtdIns(3,4,5)P3 to pleckstrin homology (PH) domains. Here we show that Ins(1,3,4,5,6)P5 inhibits the serine phosphorylation and the kinase activity of Akt/PKB. As a consequence of this inhibition, Ins(1,3,4,5,6)P5 induces apoptosis in ovarian, lung and breast cancer cells. Overexpression of constitutively active Akt protects SKBR-3 cells from Ins(1,3,4,5,6)P5-induced apoptosis. Furthermore, Ins(1,3,4,5,6)P5 enhances the proapoptotic effect of cisplatin and etoposide in ovarian and lung cancer cells, respectively. These results support a role for Ins(1,3,4,5,6)P5 as a specific inhibitor of the PI 3-K/Akt signalling pathway, that may sensitize cancer cells to the action of commonly used anticancer drugs. PMID- 14755255 TI - US visas: Postdocs in peril. PMID- 14755254 TI - IL-3 affects endothelial cell-mediated smooth muscle cell recruitment by increasing TGF beta activity: potential role in tumor vessel stabilization. AB - Interleukin-3 (IL-3) expression by tumor-infiltrating lymphocytes (TILs) and its effects on vessel assembly were evaluated. TILs from 'in situ' human breast cancers expressed CD4/CD25 antigens and IL-3. An injection of Matrigel containing SMC and IL-3 or basic-fibroblast growth factor (bFGF) into SCID mice confirmed the neoangiogenetic effect of both factors. However, in response to IL-3, but not to bFGF, only few SMC became incorporated into the nascent vessels. To evaluate the possibility that signals emanated by the nascent vasculature in the presence of IL-3 may negatively regulate SMC recruitment, conditioned media (CM) from IL-3 treated endothelial cells (EC) or SMC were tested for their biological effects on SMC and EC. CM from IL-3-treated SMC stimulated the migration of EC. In contrast, the migration of SMC was not affected by CM from IL-3-stimulated EC; however, it was greatly enhanced by blocking transforming growth factor beta (TGF beta) activity. TGF beta immunoenzymatic assay demonstrated the following: (i) the absence of TGF beta activity in CM from IL-3-stimulated EC; (ii) a barely detectable TGF beta activity in CM from IL-3-stimulated SMC; and (iii) the presence of TGF beta activity in the supernatants of SMC stimulated with CM from IL-3-, but not from bFGF-stimulated EC. Increased TGF beta mRNA expression was only detected in SMC stimulated with CM from IL-3-treated EC. Finally, the inhibitory signals induced by IL-3 in vivo were abrogated by the addition of the neutralizing TGF beta antibody. Thus, the positive immunostaining for IL-3 by TILs in 'in situ' breast cancers sustains the possibility that early in tumor development, IL-3 can contribute to the chronic immaturity of these vessels. PMID- 14755259 TI - Model droplets. PMID- 14755257 TI - Nanocluster magic. PMID- 14755260 TI - Pushing towards the digital storage limit. PMID- 14755261 TI - Reaching beyond discovery. PMID- 14755262 TI - Shrinking optical fibres. PMID- 14755264 TI - Scaling cell biology: all systems go! PMID- 14755266 TI - Nuclear transport erupts on the slopes of Mount Etna. AB - Nuclear pore complexes (NPCs) mediate the active transport of large substrates and allow the passive diffusion of small molecules into the nucleus of eukaryotic cells. The EMBO Workshop on the Mechanisms of Nuclear Transport focused on NPCs and on the soluble nucleocytoplasmic transport machinery. This meeting, organized by Valerie Doye (Institut Curie, Paris) and Ed Hurt (University of Heidelberg), was held within view of Mount Etna at Taormina, Sicily (November 1-5, 2003). Presentations emphasized the dynamic properties of the nuclear trafficking machinery, and demonstrated the continuity of nuclear transport with processes in the nucleus and cytoplasm. PMID- 14755267 TI - Keeping cartographers busy. PMID- 14755268 TI - Modifying NEMO. PMID- 14755270 TI - The heat is on with TAC1. PMID- 14755271 TI - Kinetochore sub-structure comes to MIND. PMID- 14755272 TI - An outer hair cell of the inner ear. PMID- 14755273 TI - Senescing human cells and ageing mice accumulate DNA lesions with unrepairable double-strand breaks. AB - Humans and animals undergo ageing, and although their primary cells undergo cellular senescence in culture, the relationship between these two processes is unclear. Here we show that gamma-H2AX foci (gamma-foci), which reveal DNA double strand breaks (DSBs), accumulate in senescing human cell cultures and in ageing mice. They colocalize with DSB repair factors, but not significantly with telomeres. These cryptogenic gamma-foci remain after repair of radiation-induced gamma-foci, suggesting that they may represent DNA lesions with unrepairable DSBs. Thus, we conclude that accumulation of unrepairable DSBs may have a causal role in mammalian ageing. PMID- 14755274 TI - Drugs in crops--the unpalatable truth. PMID- 14755275 TI - Biotech holds steady in Q4. PMID- 14755276 TI - China pushes liver proteomics. PMID- 14755277 TI - Genomics companies shop around for chemical expertise. PMID- 14755278 TI - Rogan steps down as USPTO director. PMID- 14755280 TI - Make or break for costimulatory blockers. PMID- 14755282 TI - Beyond the Petri dish. PMID- 14755281 TI - Questions linger over European GM food regulations. PMID- 14755283 TI - The prevalence of patent interferences in gene technology. PMID- 14755285 TI - A regulatory patchwork--human ES cell research oversight. PMID- 14755286 TI - Breaching the skin's barrier to drugs. PMID- 14755287 TI - Phage-inspired antibiotics? PMID- 14755288 TI - Enlightened receptor dynamics. PMID- 14755289 TI - The fate of transgenes in the human gut. PMID- 14755290 TI - Label-free detection becomes crystal clear. PMID- 14755293 TI - Genetic sequences: how are they patented? PMID- 14755292 TI - The HUPO PSI's molecular interaction format--a community standard for the representation of protein interaction data. AB - A major goal of proteomics is the complete description of the protein interaction network underlying cell physiology. A large number of small scale and, more recently, large-scale experiments have contributed to expanding our understanding of the nature of the interaction network. However, the necessary data integration across experiments is currently hampered by the fragmentation of publicly available protein interaction data, which exists in different formats in databases, on authors' websites or sometimes only in print publications. Here, we propose a community standard data model for the representation and exchange of protein interaction data. This data model has been jointly developed by members of the Proteomics Standards Initiative (PSI), a work group of the Human Proteome Organization (HUPO), and is supported by major protein interaction data providers, in particular the Biomolecular Interaction Network Database (BIND), Cellzome (Heidelberg, Germany), the Database of Interacting Proteins (DIP), Dana Farber Cancer Institute (Boston, MA, USA), the Human Protein Reference Database (HPRD), Hybrigenics (Paris, France), the European Bioinformatics Institute's (EMBL-EBI, Hinxton, UK) IntAct, the Molecular Interactions (MINT, Rome, Italy) database, the Protein-Protein Interaction Database (PPID, Edinburgh, UK) and the Search Tool for the Retrieval of Interacting Genes/Proteins (STRING, EMBL, Heidelberg, Germany). PMID- 14755297 TI - Graduate education and employment opportunities in bioethics. PMID- 14755298 TI - Global approaches for a healthy world. PMID- 14755299 TI - The code, the text and the language of God. When explaining science and its implications to the lay public, metaphors come in handy. But their indiscriminant use could also easily backfire. PMID- 14755300 TI - The importance of being inventive. Critics claim that patents on genes may lead to very broad monopolies and inhibit further innovation. The European patent system provides efficient legal means to address these issues. PMID- 14755301 TI - Your personal passport. With security concerns on the increase, governments are measuring the effectiveness of biometrics as a means of secure identification. But can biometrics really deliver what governments are promising? PMID- 14755302 TI - One problem at a time. Building research capacities in developing countries is necessary for economic success in the long term. But the numerous problems in doing so have to be solved for each country individually. PMID- 14755303 TI - Genomics and health care. How genomics medicine is translated into better health care largely depends on how physicians handle this information. PMID- 14755306 TI - Finding needles in a haystack. In silico identification of eukaryotic selenoprotein genes. PMID- 14755307 TI - Peroxisome proliferator-activated receptor-gamma: too much of a good thing causes harm. AB - The nuclear receptor peroxisome proliferator-activated receptor-gamma (PPARgamma) helps to translate 'what you eat' into 'what you are' because it allows dietary fatty acids (PPARgamma ligands) to modulate gene transcription. Treatments for diabetes include PPARgamma activators, as they sensitize the body to insulin. Our understanding of PPARgamma function has recently been enhanced by a flurry of human and mouse genetic studies, and the characterization of new PPARgamma ligands. This insight has led us to propose that modulating PPARgamma activity, rather than activating it, might be the most effective strategy for treating metabolic disorders, as this will improve glucose homeostasis while preventing adipogenesis. PMID- 14755312 TI - Low-dose alemtuzumab (Campath) in myeloablative allogeneic stem cell transplantation for CD52-positive malignancies: decreased incidence of acute graft-versus-host-disease with unique pharmacokinetics. AB - Alemtuzumab is effective in reducing the risk of acute graft-versus-host disease (GVHD) after allogeneic stem cell transplantation (ASCT). Alemtuzumab may also delay immune reconstitution and reduce graft-versus-leukemia effects. The optimal dose has not been established. We investigated engraftment, acute GVHD incidence and severity, and pharmacokinetics of alemtuzumab associated with the use of low dose alemtuzumab/cyclophosphamide/total body irradiation and ASCT for patients with aggressive CD52-positive hematologic malignancies. In all, 12 patients were treated. Alemtuzumab 10 mg daily on days -7 to -3 was given intravenously. Tacrolimus and methotrexate were used for GVHD prophylaxis. Alemtuzemab was not detected in any of the 36 sequential serum samples tested between days -1 and +21 of transplant. All patients engrafted rapidly; the median time to an absolute neutrophil count >0.5 x 10(9)/l was 14 days (range 11-17 days), and the median time to a platelet count >20 x 10(9)/l was 16 days (range 6-30 days). By 1 month after transplant, nine patients had 100% donor chimerism, while three had mixed donor chimerism. At 3 months, 11 had achieved 100% donor chimerism. No cases of grade III/IV acute GVHD occurred. At a median follow-up interval of 14.7 months (range 4-24), seven patients remained alive, and five remained free of disease. PMID- 14755308 TI - Biological activities of HAP46/BAG-1. The HAP46/BAG-1 protein: regulator of HSP70 chaperones, DNA-binding protein and stimulator of transcription. AB - HAP46/BAG-1M and its isoforms affect the protein-folding activities of mammalian HSP70 chaperones. They interact with the ATP-binding domain of HSP70 or HSC70, leaving the substrate-binding site available for further interactions. Trimeric complexes can therefore form with, for example, transcription factors. Moreover, HAP46/BAG-1M and the larger isoform HAP50/BAG-1L bind to DNA non-specifically and enhance transcription in vitro and upon overexpression in intact cells. These factors are linked to positive effects on cell proliferation and survival. This review focuses on DNA-binding activity and transcriptional stimulation by HAP46/BAG-1M, and presents a molecular model for the underlying mechanism. It is proposed that transcription factors are recruited into complexes with HAP46/BAG 1M or HAP50/BAG-1L through HSP70/HSC70 and that response-element-bound complexes that contain HAP46/BAG-1M and/or HAP50/BAG-1L along with HSP70s target and affect the basal transcription machinery. PMID- 14755311 TI - Allogeneic bone marrow transplantation in children with myelodysplastic syndrome or juvenile myelomonocytic leukemia: the Seattle experience. AB - The purpose of this study was to evaluate the role of allogeneic bone marrow transplantation (BMT) in children with myelodysplastic syndrome (MDS). In total, 94 consecutive pediatric patients with MDS received an allogeneic BMT from 1976 to 2001 for refractory anemia (RA) (n=25), RA with ringed sideroblasts (RARS) (n=2), RA with excess blasts (RAEB) (n=20), RAEB in transformation (RAEB-T) (n=14), juvenile myelomonocytic leukemia (JMML) (n=32) or chronic myelomonocytic leukemia (CMML) (n=1). The estimated 3-year probabilities of survival, event-free survival (EFS), nonrelapse mortality and relapse were 50, 41, 28 and 29%, respectively. Patients with RA/RARS had an estimated 3-year survival of 74% compared to 68% in those with RAEB and 33% in patients with JMML/CMML. In multivariable analysis, patients with RAEB-T or JMML were 3.9 and 3.7 times more likely to die compared to those with RA/RARS and RAEB (P=0.005 and 0.004, respectively). Patients with RAEB-T were 5.5 times more likely to relapse (P=0.01). The median follow-up among the 43 surviving patients is 10 years (range 1-25). We conclude that allogeneic BMT for children with MDS is well tolerated and can be curative. PMID- 14755313 TI - Autologous hematopoietic stem cell transplantation for refractory antiphospholipid syndrome causing myocardial necrosis. AB - Autologous hematopoietic stem cell transplantation (HSCT) is currently being evaluated as a treatment for autoimmune diseases, including systemic lupus erythematosus (SLE), that are associated with a very severe prognosis. We describe a 27-year-old woman with SLE with a 10-year history of refractory antiphospholipid syndrome (APS). She developed progressive myocardial necrosis despite treatment with corticosteroids, cyclophosphamide (CYC), cyclosporine, and immunopheresis. After conditioning with CYC, fludarabine, and antithymocyte globulin, autologous HSCT using CD34(+) selection was performed. After transplantation, the clinical symptoms caused by APS remitted, and the serum anticardiolipin antibody level decreased. Remission has persisted for 21 months after transplantation. Although a longer follow-up is required for the assessment of efficacy, autologous HSCT may cure patients with refractory APS. PMID- 14755315 TI - Failure of trilineage blood cell reconstitution after initial neutrophil engraftment in patients undergoing allogeneic hematopoietic cell transplantation frequency and outcomes. AB - The outcomes of patients who experience the failure to reconstitute a trilineage of blood cells after initial neutrophil engraftment were evaluated in 178 patients with hematologic disorders, who underwent allogeneic HCT. Of 165 qualified patients (five with primary engraftment failure; eight deaths before day 60 of HCT), 43 (26%) satisfied the criteria for the initial (n=22; failure of platelet >20 000/microl or red blood cell transfusion independence/reticulocyte count >/=1.0% by day 60) or subsequent (n=21, ANC <500/microl for >/=3 days, platelet <20 000/microl for >/=7 days, or red blood cells transfusion/reticulocyte <1.0% after initial trilineage reconstitution) failure. GVHD was the most common clinical condition associated with cytopenia (n=24). In all, 20 patients (47%) recovered at least partially with a median of 52 days (range 8-323) later, with 12 of those 20 patients recovering completely. The eventual reconstitution failure rate was 14% (23/163 patients). The number of cell lineages involved in the cytopenia was the only independent variable that predicted partial recovery (1 lineage vs 2-3 lineages with odds ratio of 8.69 (95% CI 1.96-38.60), P=0.004). Five/20 patients with vs 20/23 patients without partial recovery died. Trilineage reconstitution failures after allogeneic HCT need systematic analysis in the future studies. PMID- 14755316 TI - Incidence, outcome, and risk factors of late-onset noninfectious pulmonary complications after unrelated donor stem cell transplantation. AB - We evaluated the incidence, the risk factors, and the outcome of late-onset noninfectious pulmonary complications (LONIPCs) among 50 patients who underwent allogeneic stem cell transplantation from unrelated donors. Of the 39 patients surviving at least 3 months, 10 (26%) fulfilled the diagnostic criteria of LONIPCs and were further subclassified as having bronchiolitis obliterans (four patients), bronchiolitis obliterans with organizing pneumonia (four patients), and interstitial pneumonia (two patients). Two patients had a durable partial remission after treatment with prednisone and cyclosporine; the remaining eight patients did not respond to treatment and five of them died of respiratory failure. Advanced stage of disease at transplant and chronic extensive graft versus-host disease (GVHD) were significantly associated with the development of LONIPCs. Pulmonary function test (PFT) results before transplantation were similar in all patients, but patients with LONIPCs had a significant decrease in PFT indexes at the third month after BMT compared with controls. Moreover, the rate of cyclosporine taper during the fourth and fifth months after BMT was significantly more rapid in patients with LONIPCs than in controls, suggesting that the risk of LONIPCs may be influenced by a faster reduction of GVHD prophylaxis. PMID- 14755314 TI - Pharmacokinetics and effects on bowel and throat microflora of oral levofloxacin as antibacterial prophylaxis in neutropenic patients with haematological malignancies. AB - Gram-positive breakthrough infections pose a major drawback to the use of quinolones for antibacterial prophylaxis in neutropenic patients. Levofloxacin offers the advantage of an augmented Gram-positive spectrum and may potentially overcome this problem. In an open-label, clinical pilot study, we investigated the effects on throat and bowel microflora and pharmacokinetics of a once-daily oral dose of 500 mg levofloxacin, during neutropenia in 20 patients with haematological malignancies. Gram-negative bowel flora and Staphylococcus aureus were successfully eradicated. No Gram-negative infections occurred. Minimal inhibitory concentration values for viridans group (VG) streptococci tended to increase, in four patients over 8 mg/l, indicating resistance to levofloxacin. Four patients developed blood-stream infections with levofloxacin-resistant Gram positive cocci. No significant changes in numbers of anaerobic microorganisms were observed. Pharmacokinetic parameters of levofloxacin, including the maximum serum concentration (C(max)), time to C(max) (T(max)), area under the concentration-time curve (AUC), volume of distribution at steady state (V(ss)/F) and clearance (CL/F) were not statistically different at first dose and during neutropenia. In conclusion, levofloxacin eradicates Gram-negative microorganisms and S. aureus and spares the anaerobic flora. Its pharmacokinetic profile is unaltered during neutropenia. However, prolonged administration of levofloxacin as antibacterial prophylaxis may be hampered by the emergence of levofloxacin resistant VG streptococci. PMID- 14755317 TI - Feasibility of reduced intensity hematopoietic stem cell transplantation from an HLA-matched unrelated donor. AB - To evaluate the feasibility of reduced intensity stem cell transplantation (RIST) with bone marrow from a matched unrelated donor (MUD), we retrospectively investigated 20 patients with hematological disorders who received RIST in the Tokyo SCT consortium from January 2000 to October 2002. The preparative regimens were fludarabine-based (150-180 mg/m(2), n=18) or cladribine-based (0.77 mg/kg, n=2). To enhance engraftment, antithymocyte globulin (ATG) and 4 or 8 Gy total body irradiation (TBI) were added to these regimens in nine and 11 patients, respectively. GVHD prophylaxis was cyclosporine with or without methotrexate. In all, 19 achieved primary engraftment. Three developed graft failure (one primary, two secondary), and five died of treatment-related mortality within 100 days of transplant. Seven of the 19 patients who achieved initial engraftment developed grade II-IV acute GVHD, and seven of 13 patients who survived >100 days developed chronic GVHD. At a median follow-up of 5.5 months, estimated 1-year overall survival was 35%. Compared with a TBI-containing regimen, an ATG-containing regimen was associated with a high risk of graft failure (30 vs 0%, P=0.0737). This study supports the feasibility of RIST from MUD; however, procedure-related toxicities remain significant in its application to patients. PMID- 14755318 TI - Antimicrobial prophylaxis in hematopoietic stem cell transplant recipients: heterogeneity of current clinical practice. AB - Antimicrobial agents are commonly used after hematopoietic stem cell transplant (HSCT) to prevent bacterial, viral and fungal infections. A pharmacy practice survey was undertaken to evaluate prevailing practices. The 31 centers evaluated transplanted over 3400 patients in 2001. Over half used bacterial prophylaxis; all with fluoroquinolones. A significantly higher proportion (90-100%) used fungal and viral prophylaxis. Most centers used fluconazole for fungal prophylaxis, but the dose used varied from 400 mg (the recommended dose) to 100 mg. Itraconazole and amphotericin preparations were used by some centers for allograft recipients because of their activity against aspergillosis. Most centers used brief viral prophylaxis for autograft recipients aimed at preventing HSV reactivation. Viral prophylaxis for allograft recipients was usually much more prolonged, reflecting concern over cytomegalovirus infections. Overall, there was significant deviation from recommended guidelines in many of the practices. Our survey suggests that substantial variation exists among transplant centers in their approach to antimicrobial prophylaxis after HSCT. This probably stems from the lack of definitive studies and strong recommendations in several areas, availability of newer agents that have not been adequately studied in the HSCT setting, and a desire to improve outcome before definitive studies are available for newer agents, a process that could take several years. PMID- 14755319 TI - 'Agglutination and flocculation' of stem cells collected by apheresis due to cryofibrinogen. AB - Collection of peripheral stem cells by apheresis is a well-described process. Here, investigations concerning 'agglutination and flocculation' of stem cells collected from two patients are described. In both cases, cryoproteins were observed and cryofibrinogen was identified using high-resolution two-dimensional electrophoresis. In one case, peripheral stem cells were collected after a second course of mobilization, and the cells were immediately washed at 37 degrees C before being frozen, allowing their use, despite the presence of cryofibrinogen. In the other case, 'agglutination' was reversed by warming the bag, and plasma was removed before freezing. PMID- 14755320 TI - Donor lymphocyte infusions for refractory pure red cell aplasia relapsing after both autologous and nonmyeloablative allogeneic peripheral stem cell transplantation. AB - Pure red cell aplasia (PRCA) is characterized by a selective marrow aplasia of the erythroid compartment. Immunosuppressive therapy achieves good results in about 25% of cases, but relapses are frequent. Autologous or allogeneic haematopoietic stem cell transplantation (HSCT) may be valuable in selected patients. Here, we report details of a 29-year-old woman treated successfully by donor lymphocyte infusions (DLIs) following allogeneic HSCT for acquired refractory relapsed PRCA. The nonmyeloablative conditioning regimen consisted of cyclophosphamide 60 mg/kg/day for 2 days and fludarabine 30 mg/m(2) daily for 4 days. Haematopoiesis was still completely 'recipient' 1 month after allo-HSCT, but progressed to full donor engraftment after three doses of 'escalating' DLI. The possible role of a graft-versus-autoimmunity effect induced by allogeneic HSCT followed by DLI infusions in the treatment of the disease is discussed. PMID- 14755321 TI - Mobilized blood cells vs bone marrow harvest: experience compared in 171 donors with particular reference to pain and fatigue. AB - This prospective study compared the donor experience of blood cell (BC) mobilization and leukapheresis (n=116) with that of bone marrow (BM) harvest (n=55). Internal jugular catheters were inserted electively in 89% of BC donors. Most (80%) BM donors had a harvest with general anesthesia; 20% had epidural or spinal anesthesia. Pain and fatigue were frequent with both procedures and were compared in responses to questionnaires. A total of 85% of BM donors reported moderate or severe pain compared with 68% of BC donors (P=0.02). The median duration of pain was 14 days for BM donors compared with 3 days after BC mobilization (P<0.0001). More BM donors had pain for more than 7 days (75% vs 0%, P<0.0001). Severe fatigue was experienced by more BM donors (49 vs 16%, P<0.0001). Fatigue lasted significantly longer in BM donors (median 11 vs 4 days, P<0.0001) and more BM donors were fatigued for more than 1 week (69 vs 0%, P<0.0001). A total of 11 donors had both BM and BC collection; seven preferred the latter. Simply considered with respect to pain and fatigue, BC donation appears better tolerated by donors. However, there are other sequelae of both influencing the acceptability for individual donors. PMID- 14755322 TI - Third EBMT/AMGEN Workshop on reduced-intensity conditioning allogeneic haemopoietic stem cell transplants (RIC-HSCT), and panel consensus. AB - The Third Workshop on reduced intensity conditioning allogeneic stem cell transplants (RIC-HSCT) was convened by EBMT/AMGEN in Zurich in February 2003. Three general issues were addressed: the age effect, graft-versus-host disease (GvHD) and donor lymphocyte infusions (DLI). An age effect is seen and has a negative impact on survival; GvHD remains an issue that needs to be addressed, and DLI were shown to be a powerful tool against minimal residual disease, but not with rapidly progressing leukemia. The role of host antigen-presenting cells in the development of acute GvHD was outlined and discussed in the animal model. The emerging importance of Epstein Barr Virus infections was also discussed. Retrospective and prospective studies from the EBMT, national groups and single institutions included over 1800 patients: the incidence of acute GvHD grade III IV was 11% extensive chronic GvHD 22% and TRM 20%. Most conditioning regimens include fludarabine in association with a reduced dose of a myeloablative agent (busulfan, TBI, thiotepa or melphalan). Most transplants were from HLA-identical siblings (77%), and used peripheral blood (90%) as a stem cell source. A consensus discussion on current evidence in RIC transplants is reported in this paper. PMID- 14755323 TI - High-dose therapy and autologous stem cell transplantation in relapsing cutaneous lymphoma. AB - Treatment of cutaneous T-cell and B-cell lymphomas is difficult and relapses are frequent. To evaluate the efficiency of high-dose therapy (HDT) and autologous stem cell transplantation (ASCT) on relapsing cutaneous lymphomas, we conducted a retrospective study of 14 patients. We investigated the clinical and histological parameters of the lymphoma, previous treatments to ASCT, short-term complications of ASCT, and occurrence of a relapse. There were 11 males and three females, with a median age of 42 years. Most often, the skin disease was disseminated without extracutaneous involvement. Four patients had a B-cell lymphoma and 10 a T-cell lymphoma. CD30 was negative in 8/10 T-cell lymphomas. Before ASCT, 13 patients had chemosensitive disease; one had refractory disease. The conditioning regimen included TBI in nine cases. No toxic death occurred. Relapse of the lymphoma occurred in eight cases (T-cell lymphoma in seven cases), within 4 months after ASCT in six cases. Relapses were treated with local treatment, interferon or classical chemotherapy. At the end of the study, 11 patients were alive and three patients had died. HDT and ASCT do not benefit patients with T-cell lymphomas. For patients with disseminated relapsing cutaneous B-cell lymphomas, this procedure should be considered. PMID- 14755324 TI - A myeloablative allograft after rejection of two consecutive nonmyeloablative transplants from two different HLA identical siblings. AB - A 55-year-old female with standard risk AML in second CR received an allogenic transplant from an HLA-matched sibling, using a nonmyeloablative conditioning regimen (NMST). On day +139, she rejected her graft with autologous reconstitution. She received a second NMST from a different HLA-matched sibling with an identical conditioning regimen and immunosuppression. On day +110, she rejected the second graft, with autologous reconstitution with blasts. She received a third allograft from the first sibling with a myeloablative busulfan based conditioning regimen. She is now day +270, in CR, with full donor chimerism. PMID- 14755325 TI - An inauspicious start for the US National Biospecimen Network. PMID- 14755327 TI - The human, societal, and scientific legacy of cholera. AB - The recent history of research on cholera illustrates the importance of establishing research and care facilities equipped with advanced technologies at locations where specific health problems exist. It is in such settings, where scientific research is often considered difficult due to poverty and the lack of essential infrastructure, that investigators from many countries are able to make important advances. On this, the 25th anniversary of the founding of the International Centre for Diarrhoeal Disease Research, Bangladesh (ICDDR,B), this article seeks to recount the Centre's demonstration of how high-quality research on important global health issues, including cholera, can be accomplished in conditions that may be considered by many as unsuitable for scientific research. PMID- 14755328 TI - Central role of the P2Y12 receptor in platelet activation. AB - Platelet activation occurs in response to vessel injury and is important for the arrest of bleeding. Platelet activation during disease states leads to vascular occlusion and ischemic damage. The P2Y(12) receptor, activated by ADP, plays a central role in platelet activation and is the target of P2Y(12) receptor antagonists that have proven therapeutic value. PMID- 14755326 TI - Helicobacter pylori persistence: biology and disease. AB - Helicobacter pylori are bacteria that have coevolved with humans to be transmitted from person to person and to persistently colonize the stomach. Their population structure is a model for the ecology of the indigenous microbiota. A well-choreographed equilibrium between bacterial effectors and host responses permits microbial persistence and health of the host but confers risk of serious diseases, including peptic ulceration and gastric neoplasia. PMID- 14755329 TI - Achieving antigen-specific immune regulation. AB - A study in this issue of the JCI shows that in response to autoantigens consisting of peptides from normal proteins, patients with diabetes mount a T cell response characterized by production of IFN-gamma (see the related article beginning on page 451). However, in response to these same antigens, T cells from normal control subjects produce IL-10. The antigen-specific response characterized by release of a regulatory cytokine suggests a mechanism for the control of autoimmunity that is initiated at the time of antigen presentation. PMID- 14755330 TI - How do mutations in lamins A and C cause disease? AB - Mutations in lamins A and C, nuclear intermediate-filament proteins in nearly all somatic cells, cause a variety of diseases that primarily affect striated muscle, adipocytes, or peripheral nerves or cause features of premature aging. Two new studies (see the related articles beginning on pages 357 and 370) use lamin A/C deficient mice, which develop striated muscle disease, as a model to investigate pathogenic mechanisms. These reports provide evidence for a stepwise process in which mechanically stressed cells first develop chromatin and nuclear envelope damage and then develop secondary alterations in the transcriptional activation of genes in adaptive and protective pathways. PMID- 14755331 TI - Nitrate tolerance, oxidative stress, and mitochondrial function: another worrisome chapter on the effects of organic nitrates. AB - A shortcoming in the clinical use of organic nitrates is the development of tolerance. Recent data have suggested that the denitrification of organic nitrates is mediated by mitochondrial aldehyde dehydrogenase and that dysfunction of this enzyme is an important cause of tolerance. In this issue of the JCI, evidence in support of this hypothesis is presented in an in vivo model of nitrate tolerance (see the related article beginning on page 482). PMID- 14755332 TI - Tandem mass spectrometry in discovery of disorders of the metabolome. AB - Genetic disorders of amino acid and fatty acid metabolism can be detected with tandem mass spectrometry (MS/MS). MS/MS screening of mice subjected to chemical mutagenesis (see the related article beginning on page 434) defined a new disorder of branched-chain amino acid metabolism resembling human maple syrup urine disease. This approach has general application to the discovery of gene function in developmental and metabolic disorders. PMID- 14755333 TI - Defects in nuclear structure and function promote dilated cardiomyopathy in lamin A/C-deficient mice. AB - Laminopathies are a group of disorders caused by mutations in the LMNA gene that encodes the nuclear lamina proteins, lamin A and lamin C; their pathophysiological basis is unknown. We report that lamin A/C-deficient (Lmna(-/ )) mice develop rapidly progressive dilated cardiomyopathy (DCM) characterized by left ventricular (LV) dilation and reduced systolic contraction. Isolated Lmna(-/ ) myocytes show reduced shortening with normal baseline and peak amplitude of Ca(2+) transients. Lmna(-/-) LV myocyte nuclei have marked alterations of shape and size with central displacement and fragmentation of heterochromatin; these changes are present but less severe in left atrial nuclei. Electron microscopy of Lmna(-/-) cardiomyocytes shows disorganization and detachment of desmin filaments from the nuclear surface with progressive disruption of the cytoskeletal desmin network. Alterations in nuclear architecture are associated with defective nuclear function evidenced by decreased SREBP1 import, reduced PPARgamma expression, and a lack of hypertrophic gene activation. These findings suggest a model in which the primary pathophysiological mechanism in Lmna(-/-) mice is defective force transmission resulting from disruption of lamin interactions with the muscle-specific desmin network and loss of cytoskeletal tension. Despite severe DCM, defects in nuclear function prevent Lmna(-/-) cardiomyocytes from developing compensatory hypertrophy and accelerate disease progression. PMID- 14755334 TI - Lamin A/C deficiency causes defective nuclear mechanics and mechanotransduction. AB - Mutations in the lamin A/C gene (LMNA) cause a variety of human diseases including Emery-Dreifuss muscular dystrophy, dilated cardiomyopathy, and Hutchinson-Gilford progeria syndrome. The tissue-specific effects of lamin mutations are unclear, in part because the function of lamin A/C is incompletely defined, but the many muscle-specific phenotypes suggest that defective lamin A/C could increase cellular mechanical sensitivity. To investigate the role of lamin A/C in mechanotransduction, we subjected lamin A/C-deficient mouse embryo fibroblasts to mechanical strain and measured nuclear mechanical properties and strain-induced signaling. We found that Lmna-/- cells have increased nuclear deformation, defective mechanotransduction, and impaired viability under mechanical strain. NF-kappaB-regulated transcription in response to mechanical or cytokine stimulation was attenuated in Lmna-/- cells despite increased transcription factor binding. Lamin A/C deficiency is thus associated with both defective nuclear mechanics and impaired mechanically activated gene transcription. These findings suggest that the tissue-specific effects of lamin A/C mutations observed in the laminopathies may arise from varying degrees of impaired nuclear mechanics and transcriptional activation. PMID- 14755335 TI - Glycoprotein 130 regulates bone turnover and bone size by distinct downstream signaling pathways. AB - The gp130-dependent cytokines, which signal through at least two intracellular pathways, regulate osteoclast and osteoblast formation. To define their roles in regulating bone mass, we analyzed mice in which gp130 signaling via either the signal transducer and activator of transcription (STAT) 1/3 (gp130(DeltaSTAT/DeltaSTAT)) or SHP2/ras/MAPK (gp130(Y757F/Y757F)) pathway was attenuated. In gp130(DeltaSTAT/DeltaSTAT) mice, trabecular bone volume (BV/TV) and turnover were normal, but bone length was reduced by premature growth plate closure, indicating an essential role for gp130-STAT1/3 signaling in chondrocyte differentiation. In contrast, while bone size was normal in gp130(Y757F/Y757F) mice, BV/TV was reduced due to high bone turnover, indicated by high osteoclast surface/bone surface (OcS/BS) and osteoblast surface/bone surface (ObS/BS). Furthermore, generation of functional osteoclasts from bone marrow of gp130(Y757F/Y757F) mice was elevated, revealing that while gp130 family cytokines stimulate osteoclastogenesis through the osteoblast lineage, gp130, via SHP2/Ras/MAPK, inhibits osteoclastogenesis in a cell lineage-autonomous manner. Genetic ablation of IL-6 in gp130(Y757F/Y757F) mice exacerbated this osteopenia by reducing ObS/BS without affecting OcS/BS. Thus, while IL-6 is critical for high bone formation in gp130(Y757F/Y757F) mice, it is not involved in the increased osteoclastogenesis. In conclusion, gp130 is essential for normal bone growth and trabecular bone mass, with balanced regulation depending on selective activation of STAT1/3 and SHP2/ras/MAPK, respectively. Furthermore, the latter pathway can directly inhibit osteoclastogenesis in vivo. PMID- 14755336 TI - Extensive tissue-regenerative capacity of neonatal human keratinocyte stem cells and their progeny. AB - Given our recent discovery that it is possible to separate human epidermal stem cells of the skin from their more committed progeny (i.e., transit-amplifying cells and early differentiating cells) using FACS techniques, we sought to determine the comparative tissue regeneration ability of these keratinocyte progenitors. We demonstrate that the ability to regenerate a fully stratified epidermis with appropriate spatial and temporal expression of differentiation markers in a short-term in vitro organotypic culture system is an intrinsic characteristic of both epidermal stem and transit-amplifying cells, although the stem cell fraction is most capable of achieving homeostasis. Early differentiating keratinocytes exhibited limited short-term tissue regeneration under specific experimental conditions in this assay, although significant improvement was obtained by manipulating microenvironmental factors, that is, coculture with minimally passaged dermal cells or exogenous supply of the ECM protein laminin-10/11. Importantly, transplantation of all classes of keratinocyte progenitors into an in vivo setting demonstrated that tissue regeneration can be elicited from stem, transit-amplifying, and early differentiating keratinocytes for up to 10 weeks. These data illustrate that significant proliferative and tissue-regenerative capacity resides not only in keratinocyte stem cells as expected, but also in their more committed progeny, including early differentiating cells. PMID- 14755337 TI - TNF-alpha is a critical negative regulator of type 1 immune activation during intracellular bacterial infection. AB - TNF-alpha has long been regarded as a proimmune cytokine involved in antimicrobial type 1 immunity. However, the precise role of TNF-alpha in antimicrobial type 1 immunity remains poorly understood. We found that TNF-alpha deficient (TNF(-/-)) mice quickly succumbed to respiratory failure following lung infection with replication-competent mycobacteria, because of apoptosis and necrosis of granuloma and lung structure. Tissue destruction was a result of an uncontrolled type 1 immune syndrome characterized by expansion of activated CD4 and CD8 T cells, increased frequency of antigen-specific T cells, and overproduction of IFN-gamma and IL-12. Depletion of CD4 and CD8 T cells decreased IFN-gamma levels, prevented granuloma and tissue necrosis, and prolonged the survival of TNF(-/-) hosts. Early reconstitution of TNF-alpha by gene transfer reduced the frequency of antigen-specific T cells and improved survival. TNF alpha controlled type 1 immune activation at least in part by suppressing T cell proliferation, and this suppression involved both TNF receptor p55 and TNF receptor p75. Heightened type 1 immune activation also occurred in TNF(-/-) mice treated with dead mycobacteria, live replication-deficient mycobacteria, or mycobacterial cell wall components. Our study thus identifies TNF-alpha as a type 1 immunoregulatory cytokine whose primary role, different from those of other type 1 cytokines, is to keep an otherwise detrimental type 1 immune response in check. PMID- 14755338 TI - Site and mechanism of leptin action in a rodent form of congenital lipodystrophy. AB - Lipodystrophy is characterized by the complete or partial absence of adipose tissue, insulin resistance, hepatic steatosis, and leptin deficiency. Here, we show that low-dose central leptin corrects the insulin resistance and fatty liver of lipodystrophic aP2-nSREBP-1c mice, while the same dose given peripherally does not. Central leptin also repressed stearoyl-CoA desaturase-1 (SCD-1) RNA and enzymatic activity, which were increased in livers of lipodystrophic mice. aP2 nSREBP-1c mice homozygous for an SCD-1 deletion had markedly reduced hepatic steatosis, increased saturated fatty acids, decreased acetyl-CoA carboxylase activity, and decreased malonyl-CoA levels in the liver. Despite the reduction in hepatic steatosis, these mice remained diabetic. A leptin dose-response curve showed that subcutaneous leptin improved hyperglycemia and hyperinsulinemia in aP2-nSREBP-1c mice at doses that did not substantially alter hepatic steatosis or hepatic SCD enzymatic activity. Leptin treatment at this dose improved insulin stimulated insulin receptor and insulin receptor substrate 2 (IRS-2) phosphorylation, IRS-2-associated PI3K activity, and Akt activity in liver. Together, these data suggest that CNS-mediated repression of SCD-1 contributes to leptin's antisteatotic actions. Intracerebroventricular leptin improves glucose homeostasis by improving insulin signal transduction in liver, but in this case the effect appears to be independent of SCD-1. PMID- 14755339 TI - High vaccination efficiency of low-affinity epitopes in antitumor immunotherapy. AB - Most of the human tumor-associated antigens (TAAs) characterized thus far are derived from nonmutated "self"-proteins. Numerous strategies have been developed to break tolerance to TAAs, combining various forms of antigens with different vectors and adjuvants. However, no study has yet determined how to select epitopes within a given TAA to induce the highest antitumor effector response. We addressed this question by evaluating in HLA-A*0201-transgenic HHD mice the antitumor vaccination efficacy of high- and low-affinity epitopes from the naturally expressed murine telomerase reverse transcriptase (mTERT). Immunity against low-affinity epitopes was induced with heteroclitical variants. We show here that the CTL repertoire against high-affinity epitopes is partially tolerized, while that against low-affinity epitopes is composed of frequent CTLs with high avidity. The high-affinity p797 and p545 mTERT epitopes are not able to protect mice from a lethal challenge with the mTERT-expressing EL4-HHD tumor. In contrast, mice developing CTL responses against the p572 and p988 low-affinity epitopes exhibit potent antitumor immunity and no sign of autoimmune reactivity against TERT-expressing normal tissues. Our results strongly argue for new TAA epitope selection and modification strategies in antitumor immunotherapy applications in humans. PMID- 14755340 TI - ENU mutagenesis identifies mice with mitochondrial branched-chain aminotransferase deficiency resembling human maple syrup urine disease. AB - Tandem mass spectrometry was applied to detect derangements in the pathways of amino acid and fatty acid metabolism in N-ethyl-N-nitrosourea-treated (ENU treated) mice. We identified mice with marked elevation of blood branched-chain amino acids (BCAAs), ketoaciduria, and clinical features resembling human maple syrup urine disease (MSUD), a severe genetic metabolic disorder caused by the deficiency of branched-chain alpha-keto acid dehydrogenase (BCKD) complex. However, the BCKD genes and enzyme activity were normal. Sequencing of branched chain aminotransferase genes (Bcat) showed no mutation in the cytoplasmic isoform (Bcat-1) but revealed a homozygous splice site mutation in the mitochondrial isoform (Bcat-2). The mutation caused a deletion of exon 2, a marked decrease in Bcat-2 mRNA, and a deficiency in both BCAT-2 protein and its enzyme activity. Affected mice responded to a BCAA-restricted diet with amelioration of the clinical symptoms and normalization of the amino acid pattern. We conclude that BCAT-2 deficiency in the mouse can cause a disease that mimics human MSUD. These mice provide an important animal model for study of BCAA metabolism and its toxicity. Metabolomics-guided screening, coupled with ENU mutagenesis, is a powerful approach in uncovering novel enzyme deficiencies and recognizing important pathways of genetic metabolic disorders. PMID- 14755341 TI - Defects in secretion, aggregation, and thrombus formation in platelets from mice lacking Akt2. AB - Prior studies have shown that PI3Ks play a necessary but incompletely defined role in platelet activation. One potential effector for PI3K is the serine/threonine kinase, Akt, whose contribution to platelet activation was explored here. Two isoforms of Akt were detected in mouse platelets, with expression of Akt2 being greater than Akt1. Deletion of the gene encoding Akt2 impaired platelet aggregation, fibrinogen binding, and granule secretion, especially in response to low concentrations of agonists that activate the G(q) coupled receptors for thrombin and thromboxane A(2). Loss of Akt2 also impaired arterial thrombus formation and stability in vivo, despite having little effect on platelet responses to collagen and ADP. In contrast, reducing Akt1 expression had no effect except when Akt2 was also deleted. Activation of Akt by thrombin was abolished by deletion of Galpha(q) but was relatively unaffected by deletion of Galpha(i2), which abolished Akt activation by ADP. From these results we conclude that Akt2 is a necessary component of PI3K-dependent signaling downstream of G(q)-coupled receptors, promoting thrombus growth and stability in part by supporting secretion. The contribution of Akt1 is less evident except in the setting in which Akt2 is absent. PMID- 14755342 TI - Autoreactive T cell responses show proinflammatory polarization in diabetes but a regulatory phenotype in health. AB - According to the quality of response they mediate, autoreactive T cells recognizing islet beta cell peptides could represent both disease effectors in the development of type 1 diabetes (T1DM) and directors of tolerance in nondiabetic individuals or those undergoing preventative immunotherapy. A combination of the rarity of these cells, inadequate technology, and poorly defined epitopes, however, has hampered examination of this paradigm. We have identified a panel of naturally processed islet epitopes by direct elution from APCs bearing HLA-DR4. Employing these epitopes in a sensitive, novel cytokine enzyme-linked immunosorbent spot assay, we show that the quality of autoreactive T cells in patients with T1DM exhibits extreme polarization toward a proinflammatory Th1 phenotype. Furthermore, we demonstrate that rather than being unresponsive, the majority of nondiabetic, HLA-matched control subjects also manifest a response against islet peptides, but one that shows extreme T regulatory cell (Treg, IL-10-secreting) bias. We conclude that development of T1DM depends on the balance of autoreactive Th1 and Treg cells, which may be open to favorable manipulation by immune intervention. PMID- 14755343 TI - Salicylate induces an antibiotic efflux pump in Burkholderia cepacia complex genomovar III (B. cenocepacia). AB - An antibiotic efflux gene cluster that confers resistance to chloramphenicol, trimethoprim, and ciprofloxacin has been identified in Burkholderia cenocepacia (genomovar III), an important cystic fibrosis pathogen. Five open reading frames have been identified in the cluster. There is apparently a single transcriptional unit, with llpE encoding a lipase-like protein, ceoA encoding a putative periplasmic linker protein, ceoB encoding a putative cytoplasmic membrane protein, and opcM encoding a previously described outer membrane protein. A putative LysR-type transcriptional regulatory gene, ceoR, is divergently transcribed upstream of the structural gene cluster. Experiments using radiolabeled chloramphenicol and salicylate demonstrated active efflux of both compounds in the presence of the gene cluster. Salicylate is an important siderophore produced by B. cepacia complex isolates, and both extrinsic salicylate and iron starvation appear to upregulate ceoR promoter activity, as does chloramphenicol. These results suggest that salicylate is a natural substrate for the efflux pump in B. cenocepacia and imply that the environment of low iron concentration in the cystic fibrosis lung can induce efflux-mediated resistance, even in the absence of antibiotic selective pressure. PMID- 14755344 TI - Conditional disruption of IkappaB kinase 2 fails to prevent obesity-induced insulin resistance. AB - The inhibitor of NF-kappaB (IkappaB) kinases (IKK1[alpha] and IKK2[beta]), the catalytic subunits of the IKK complex, phosphorylate IkappaB proteins on serine residues, targeting them for degradation and thus activating the transcription factor NF-kappaB. More recently, IKK2 has been implicated in mediation of insulin resistance caused by obesity, lipid infusion, and TNF-alpha stimulation, since salicylate and aspirin, known inhibitors of IKK activity, can reverse insulin resistance in obese mouse models. To further genetically elucidate the role of IKK2 in obesity-mediated insulin resistance, we have conditionally inactivated the mouse IKK2 gene in adult myocytes by Cre-loxP-mediated recombination in vivo. We have investigated the development of obesity-induced insulin resistance in muscle-specific IKK2 knockout mice and mice exhibiting a 50% reduction of IKK2 expression in every tissue and have found that, after gold thioglucose treatment, wild-type and mutant mice developed obesity to a similar extent. Surprisingly, no difference in obesity-induced insulin resistance was detectable, either at a physiological or at a molecular level. Moreover, impaired glucose tolerance resulting from a high-fat diet occurred to the same degree in control and IKK2 mutant mice. These data argue against a substantial role for muscular IKK2 in mediating obesity-induced insulin resistance in these models in vivo. PMID- 14755345 TI - Central role of mitochondrial aldehyde dehydrogenase and reactive oxygen species in nitroglycerin tolerance and cross-tolerance. AB - Recent studies suggest that mitochondrial aldehyde dehydrogenase (ALDH-2) plays a central role in the process of nitroglycerin (glyceryl trinitrate, GTN) biotransformation in vivo and that its inhibition accounts for mechanism-based tolerance in vitro. The extent to which ALDH-2 contributes to GTN tolerance (impaired relaxation to GTN) and cross-tolerance (impaired endothelium-dependent relaxation) in vivo remain to be elucidated. Rats were treated for three days with GTN. Infusions were accompanied by decreases in vascular ALDH-2 activity, GTN biotransformation, and cGMP-dependent kinase (cGK-I) activity. Further, whereas in control vessels, multiple inhibitors and substrates of ALDH-2 reduced both GTN-stimulation of cGKI and GTN-induced vasodilation, these agents had little effect on tolerant vessels. A state of functional tolerance (in the GTN/cGMP pathway) was recapitulated in cultured endothelial cells by knocking down mitochondrial DNA (rho(0) cells). In addition, GTN increased the production of reactive oxygen species (ROS) by mitochondria, and these increases were associated with impaired relaxation to acetylcholine. Finally, antioxidants/reductants decreased mitochondrial ROS production and restored ALDH 2 activity. These observations suggest that nitrate tolerance is mediated, at least in significant part, by inhibition of vascular ALDH-2 and that mitochondrial ROS contribute to this inhibition. Thus, GTN tolerance may be viewed as a metabolic syndrome characterized by mitochondrial dysfunction. PMID- 14755356 TI - Infrasternal mediastinoscopic surgery for anterior mediastinal masses. AB - BACKGROUND: Infrasternal mediastinoscopic surgery is a new alternative to the thoracoscopic approach for patients with anterior mediastinal masses. METHODS: We applied this technique to 18 thymectomies, one thymomectomy, and one cystectomy in a total of 20 patients with anterior mediastinal masses and then assessed the surgical results. RESULTS: Infrasternal mediastinoscopic surgery was accomplished in 18 of the 20 patients (90%). The pathological diagnoses included 13 Masaoka stage I thymomas, one stage II thymoma, two stage III thymomas, one thymic cyst, one pericardial cyst, one thymic granuloma, and one mature teratoma. Two patients with stage III thymoma required conversion to sternotomy, one for invasion into the innominate vein and the other for invasion into the pericardium. There was no surgically related mortality or complications in any patients. CONCLUSION: Infrasternal mediastinoscopic surgery is safe and feasible for stage I thymoma and other benign tumors in the anterior mediastinum. PMID- 14755357 TI - [Probiotics]. AB - The colonic flora is thought to play a key role in human health. Gut bacteria produce desirable as well as undesirable metabolites from fermentation and sometimes they may act as pathogens. Composition as well as activities of the gut bacteria can be influenced by nutrition. Prebiotics are food ingredients that selectively stimulate the growth of colonic bacteria regarded as beneficial, i. e. bifidobacteria and lactobacilli. Fructose polymers (fructo-oligosaccharides, inulin), galacto-oligosaccharides and soya oligosaccharides were shown to exert prebiotic activities. In adults, the growth of bifidobacteria was stimulated especially by inulin and fructo-oligosaccharides. Infant formulas containing galacto-oligosaccharides and inulin promoted the growth of bifidobacteria in faeces of bottle-fed infants and resulted in stool characteristics similar to those found in breast-fed infants. However, at present there is only limited knowledge on the long-term health consequences of increased counts of bifidobacteria. In adults, prebiotics can enhance the absorption of calcium from the colon. They can increase stool frequency and are mildly laxative. Prebiotic consumption is usually accompanied by mild flatulence and in sensitive persons by more severe gastrointestinal symptoms. In contrast to the results of animal experiments, a significant lipid-lowering action of fructo-oligosaccharides has not been found in humans. In animals, fructo-oligosaccharides have shown a favourable effect on experimental carcinogenesis. Whether or not these findings may apply to humans will be subject of future studies. PMID- 14755358 TI - [Contraception in adolescents--address on the occasion of the FBA 2002]. AB - In Germany we observe an increasing number of unwanted teenage pregnancies associated with a consecutively escalating rate of abortions. The education at school, through parents and public media does not show sufficient prevention efficacy. One of the key issues in this context seems to be the discrepancy between time of education and premature sexual activity. Suitable contraceptives are required to meet individual needs and parameters of the adolescent. There are various suitable options as low dose oral contraception of the new generation, long-term depot contraceptives, the Nuva-Ring and future transdermal and subcutaneous contraceptives currently under evaluation. The interests and demands of the teenage patient are the focus of attention but the legal ground has to be taken into consideration, too: Contraception for adolescents provides potential conflict of interest for the physician and thus makes knowledge of recent laws essential. PMID- 14755359 TI - [A guideline for guidelines--methodological report and use of the guideline women's information]. AB - Information and education is needed to empower autonomy and self-determination of patients (informed consent). Furthermore reliable and accurate medical information is necessary for patients who want to take an active part in medical decision-making. The aim of this work is to define the requirements helping to assure the development of good qualified information material relevant for women and female patients as "a guideline on women information". An example of its use is given by embeding this guideline in the guideline for early detection of breast cancer in Germany by defining the specific elements required for developing qualified information on this issue for women. METHODS: A systematic, stepwise methodological process according to a level two guideline of the German Association of the Scientific Medical Societies (AWMF) and the Agency for Quality in Medicine (AZQ) was performed with the following elements: 1. Establishing an expert panel, 2. Generating the guideline statements by a formal, consensus based nominal group process, 3. External review process and finding supportive partners for the guideline on women information, 4. Using the guideline for guidelines: implementing the concept in the guideline of early detection of breast cancer in Germany. RESULTS: The "guideline women information" comprises nine elements of quality assuring requirements for the development of gender-specific information material and eleven specific elements which directly relate to the guideline statements on early detection of breast cancer. After external review 30 organisations gave their written support for future implementation of the guideline. The "guideline women information" was integrated as a tool for quality assurance of lay information into the "guideline for early detection of breast cancer in Germany". CONCLUSION: The "guideline women information" is a systematically developed, consensus-based recommendation to improve the development of qualified lay information at the point of its process by defining gender-specific aspects required for good lay information and its evaluation. As a guideline for guidelines its use is demonstrated by integrating this guideline into the "guideline for early detection of breast cancer in Germany" to ensure the development of qualified guideline compliant information. PMID- 14755360 TI - [Counselling, genetic testing and prevention in women with hereditary breast- and ovarian cancer. Interdisciplinary recommendations of the consortium "Hereditary Breast- and Ovarian Cancer" of the German Cancer AiD]. AB - Women with familial predisposition for breast and ovarian cancer represent a small group of patients with very high risk for developing breast and/ or ovarian cancer before the age of 50 years. The individual breast cancer risk can be assessed by genetic counselling and can be specified by genetic diagnostics. As part of the gynaecological consultation, adequate preventive measures are offered. Psycho-oncological counselling may help in decision making. For hereditary carcinomas, counselling is still not considered as a routine medical care, even though basic and routine preventive measures are insufficient for this group of high risk patients. Within the last years, 12 specialized centres in Germany have developed a patient care concept for women with a familial risk for breast and ovarian cancer. Establishment of these centres for familial breast and ovarian cancer and use of evidence-based medical care was initiated in a nationwide interdisciplinary joint research project and supported by the German Cancer AiD. These measures were integrated in a quality assurance concept for structure, process and result optimization. Thus, all requirements for introducing these services into routine patient management have been fulfilled. PMID- 14755361 TI - [The use of oral antidiabetic drugs in the treatment of polycystic ovary syndrome]. AB - The polycystic ovary syndrome (PCOS) is the most common endocrinopathy in women of reproductive age. It is a complex metabolic-endocrine disorder with severe long-term health consequences like type 2 diabetes. The increased risk for cardiovascular diseases in women with PCOS is due to diabetes, adipositas and dyslipidemia. Insulin resistance plays a key role in the pathophysiology of this syndrome. This makes the use of oral antidiabetic drugs most compelling. The majority of studies have shown amelioration of the typical symptoms like hyperandrogenism and cycle irregularities. Ovulation and pregnancy rates increased. Furthermore these drugs might be cardioprotective by improving insulin sensitivity and reduce the risk for type 2 diabetes. This article reviews the use of different oral antidiabetic drugs in the treatment of PCOS and their influence on fertility, the risk for type 2 diabetes and cardiovascular diseases. PMID- 14755362 TI - [Plastic surgery of defects of the thoracic wall]. AB - Defects of the thoracic wall are mostly due to a malignant disease of the mammary gland. Radiotherapy is one of the essential methods of treating cancer of the breast in addition to primary surgery. If, after surgery, healing occurs by secondary intention or if there is a local recurrence or tumour formation in the thoracic wall due to irradiation, the body's reserves of soft parts are often exhausted and the adjacent tissue has been additionally damaged by the irradiation. General principles of treatment in the management of such problems are presented by means of several case reports. PMID- 14755363 TI - Mepivacaine for spinal anesthesia in parturients undergoing elective cesarean delivery: maternal and neonatal plasma concentrations and neonatal outcome. AB - OBJECTIVE: Worldwide, long-acting bupivacaine is most commonly used for spinal anesthesia in parturients undergoing elective Cesarean delivery. However, advances in surgical technique and shorter duration of surgery make short-acting local anesthetic like mepivacaine appropriate, particularly if combined with opioids to enhance postoperative maternal pain relief. MATERIAL AND METHODS: We assessed the effect of 4% hyperbaric mepivacaine (60 mg) plus 10 microg fentanyl for spinal anesthesia in 11 parturients undergoing elective Cesarean delivery. Sensory, motor and analgesic block characteristics, neonatal outcome (Apgar scores, umbilical cord blood analysis, neurologic and adaptive capacity score) as well as fetal and maternal mepivacaine plasma concentrations at delivery were determined (HPLC/UV). RESULTS: Motor block (Induction-Bromage 0) duration lasted 113 +/- 20 min. Effective analgesia (VAS < or = 40) was 128 +/- 35 min. Maternal and fetal mepivacaine free plasma concentration were 0.18 +/- 0.05 microg/ml and 0.10 +/- 0.03 microg/ml, respectively. The fetal to maternal (UV/MV)-ratio for mepivacaine free plasma concentration was 0.56. Apgar scores, NACScores and the umbilical blood analysis showed no evidence of neonatal depression. CONCLUSIONS: Particularly with short duration of surgery (21 +/- 5 min) intrathecal mepivacaine combined with fentanyl offers a favorable clinical alternative in parturients undergoing elective Cesarean delivery. PMID- 14755364 TI - [Hemoperitoneum after early abortion of an abdominal pregnancy--reduced therapeutic options in case of delayed diagnosis]. AB - BACKGROUND: Early abortion of a abdominal pregnancy is a rare gynecological emergency occurring in 1/10(4) pregnancies in the US. In unconscious patients in the reproductive age this differential diagnosis has to be taken in mind for the choice of the therapeutic management. CASE: A 30- year-old woman (II-gravida, I para) suffering for about 18 hours from "upper abdominal pain" was found in somnolent and in reduced conditions by her relatives. Unconscious and with all signs of a hemorrhagic shock, she was transferred to the emergency department of the UKBF. After stabilization of circulation, a diagnostic laparoscopy has been performed with the finding of about 2000 ml of coagulated and fresh blood, but without the confirmation of a extrauterine pregnancy (EUP): the small pelvis and his organs were without any signs of EUP. The laparoscopic approach has been changed into a secondary median laparotomy. After exclusion of other reasons for a intraabdominal bleeding (such as spleen rupture or gastrointestinal bleeding), and the complete removal of all intraabdominal blood clots, a embryo, his placenta and the place of placental insertion has been found in the mesenterium jejuni. The surgical procedure was finished in the usual way. The postoperative healing process was uncomplicated. CONCLUSIONS: Our case did meet Veits criteria of a abdominal pregnancy: intact embryo, no contact between placenta and fallopian tubes or ovaries, resp., but definitive insertion of the placenta in the distant peritoneum. In woman in their reproductive age suffering from intraabdominal bleeding, the exclusion of a ectopic pregnancy is essential. In the case of a vital emergency, and because of suboptimal management conditions (without a sufficient patients history, lack of vaginal sonography, instable circulation because of delayed diagnosis), excluding the opportunity of a laparoscopic exploration, the correct diagnosis of the early abortion of a abdominal pregnancy has to be made by open surgery. PMID- 14755366 TI - [Cochlea implantation: results and costs, a review]. AB - The cochlear implantation, especially after bilateral operation provides a reliable and outstanding compensation in bilateral complete deafness or severe hearing impairment. Directional hearing can be restored with bilateral implantation so that warning signals are localized correctly and speech in noise is easier understood. Thus, rehabilitation is restored to a high degree. The involved expenses are severe, but are lower for the society when compared to the amount of money necessary to substitute a completely deaf patient lifelong. PMID- 14755367 TI - [Critical analysis of complications and disorders in wound healing after tracheostomy in children]. AB - BACKGROUND: The analysis of early and late postoperative results after tracheostomy in children. METHODS: A 6 year, prospective, observational cohort study was undertaken in 14 children of mean age 4 years 7 months (range 0.03 month-15 years) at the Department of Otorhinolaryngology, University of Ulm. Children were included if they had at least two follow-up examinations postoperatively because of local disorders in wound healing of the tracheostoma. Etiologic factors, the type of tracheotomy, the size and type of the tracheostomy tube, primary and secondary local disorders in wound healing of the tracheostoma, and the clinical follow-up were analyzed. RESULTS: Formation of granulation tissue and tracheal stenosis were the most observed local disorders. In five patients, the tracheal stenosis was located below the stoma, in one patient also above the stoma. A stenosis of the stoma was found in two patients. One patient died because of a tracheoinnominate artery fistula. In 10 patients problems in swallowing and recurrent aspiration of saliva was observed early postoperatively. CONCLUSIONS: The tracheotomy in childhood is often related with late wound healing disorders. An association of wound healing disorders with a feasible lethal outcome is rarely found. Local formation of granulation tissue renders decannulation more difficult. The choice of a correctly fitting tracheostomy tube and special care of the tracheostoma in long-term cannulated children has to be emphasized. PMID- 14755368 TI - [Complications of lumbar administration of 5% sodium fluorescein solution for detection of cerebrospinal fluid fistula]. AB - BACKGROUND: The detection of cerebrospinal fluid fistulas in the region of the anterior or lateral skull base can be difficult. The fluorescein test with lumbar administration of 5% sodium fluorescein solution can be used to detect cerebrospinal fluid leakage, identify weak points in the dura, achieve precise localisation of cerebrospinal fluid fistulas and to check intraoperatively that watertight dural closure has been achieved. However, use of the test is problematic as the fluorescein solution used is not licensed for this indication in Germany and severe neurological complications are described in the literature. In order to clarify the legal situation regarding use of the test, we therefore analysed the complications occurring in a sizeable patient sample. METHOD: The records of all patients in whom a fluorescein test had been performed between 1979 and June 2000 were analysed retrospectively for the occurrence of complications. RESULTS: The most frequent complication in the 368 fluorescein tests performed was headache, followed by nausea and vomiting, temperature elevation, dizziness and nuchal pain. These side-effects were no more frequent than described for lumbar puncture alone. Twenty-six patients experienced side effects on the day of the operation, 65 on the first postoperative day, 36 on the second day, 34 on the third day and 13 patients after the third day. There were two cases of grand mal seizures following concomitant intrathecal contrast medium administration. None of the patients had side effects persisting longer than 4 weeks. CONCLUSIONS: Intrathecal administration of a 5 % fluorescein solution is a safe procedure provided that the maximum dosages are not exceeded and the solution is prepared and administered correctly and in accordance with the specified indications and contraindications. In view of its great diagnostic benefit and low risk when properly used, the dictates of therapeutic freedom allow use of this drug despite the fact that it is not licensed for this purpose in Germany. It is necessary to obtain written informed consent from the patient. PMID- 14755369 TI - [Bioimplants--characteristics and use]. AB - Bioimplants are materials of human or animal origin that are offered after conservation or preparation to the tissue replacement. They are used in the different fields of ENT with very good successes. On the basis of its origin, the infection security plays a large role next to the general demands on an implant. The materials related today and its fields of application are discussed. PMID- 14755370 TI - B-CAM/LU expression and the role of B-CAM/LU activation in binding of low- and high-density red cells to laminin in sickle cell disease. AB - Red blood cells from patients with sickle cell disease (SS RBC) adhere to laminin and over-express the high-affinity laminin receptor basal cell adhesion molecule/Lutheran protein (B-CAM/LU). This receptor has recently been shown to undergo activation in vitro through a protein kinase A-dependent mechanism. Low density SS RBC express two-thirds more B-CAM/LU than high-density SS RBC. However, high-density SS RBC have been identified as most adherent to laminin under flow conditions. We investigated the ability of low- and high-density SS RBC to interact with laminin under various conditions and explored factors that might be responsible for the differences in B-CAM/LU-laminin interaction between high- and low-density SS RBC. We confirmed that high-density SS RBC adhere to laminin more strongly than low-density SS RBC under flow conditions. However, low density SS RBC bind soluble laminin most strongly and are the most adherent to laminin under static conditions. Soluble recombinant Lutheran extracellular domain protein completely blocked SS RBC adhesion to laminin under both static and flow conditions. The protein kinase A inhibitor 14-22 amide inhibited adhesion to laminin during flow by high-density SS RBC from patients with strongly adherent cells but had no effect on adhesion observed after a static phase. Deletion of the cytoplasmic domain of B-CAM as well as mutation of the juxtamembranous tyrosine residue failed to reduce B-CAM-mediated adhesion to laminin by transfected MEL cells. These studies confirm that B-CAM/LU is the most critical receptor mediating adhesion to laminin under both static and flow conditions. Dense SS RBC are most adherent to laminin despite bearing fewer laminin receptors, apparently due to a reversible protein kinase A-dependent process that is unlikely to involve direct phosphorylation of B-CAM/LU. Our results also suggest that the nature of the interaction of B-CAM/LU with laminin may be different under static and flow conditions. PMID- 14755371 TI - C1272S: a new candidate mutation in type 2A von Willebrand disease that disrupts the disulfide loop responsible for the interaction of VWF with platelet GP Ib-IX. AB - Most of type 2A von Willebrand disease (VWD) mutations are clustered within the A2 domain of VWF, encoded by the 3' region of exon 28 of the von Willebrand factor (VWF) gene. A patient with lifelong and severe bleeding diathesis and laboratory data of type 2A VWD is described. The analysis of the complete exon 28 of the VWF gene showed a 3815 G-->C change within the A1 domain, resulting in the C1272S missense mutation in a heterozygous state. The substitution was not found in 100 normal alleles also examined and has not been described previously. This candidate mutation would interrupt the formation of the disulfide loop 1272-1458, which is important in maintaining the adequate conformation of the VWF functional domain that interacts with platelet glycoprotein Ib-IX. Gene expression of this candidate mutation is necessary to confirm its role. PMID- 14755372 TI - Th1/Th2 cytokine profiles and their relationship to clinical features in patients following nonmyeloablative allogeneic stem cell transplantation. AB - An imbalance in helper T-cell type 1 (Th1) and type 2 (Th2) cytokines is suggested to play an important role in the pathogenesis of acute graft-versus host disease (aGVHD). The aim of this study was to investigate the cytokine bias acquired by T cells after transplantation and its possible influence on relapse of original malignancy. Cytokine levels by peripheral CD4 and CD8 T cells were tested at various pre- and post-transplant time points with fluorescein isothiocyanate-based intracellular cytokine assay after short-term in vitro mitogenic stimulation (phorbol myristate acetate + ionomycin). In both CD4+ and CD8+ cells, interferon (IFN)-gamma-producing cell populations increased, indicating a shift to a Th1 cytokine profile with aGVHD. IFN-gamma-producing T cells was significantly lower in patients who experienced relapse of original disease compared to those who showed no signs of relapse and compared to normal controls. Our studies demonstrate that aGVHD correlates with a Th1 bias and that Th1 response may potentiate an effective immune surveillance. PMID- 14755373 TI - Erythropoietin-deficient anemia associated with autoimmune polyglandular syndrome type I. AB - Autoimmune polyglandular syndrome type I (APS1), a relatively common disorder in some populations, is frequently associated with adrenal insufficiency, hypoparathyroidism, and other endocrine and skin abnormalities. We describe an 18 year-old male with APS1, as documented by genotyping, who presented with hypoparathyroidism and a normocytic, hypoproliferative, isolated anemia. An extensive hematological work-up revealed a low serum erythropoietin, without any other hematological abnormalities. His renal function was normal, and he did not have many of the laboratory or clinical findings associated with an anemia of chronic disease. His anemia was responsive to superphysiologic doses of erythropoietin. We thus suggest that erythropoietin deficiency may be one of the endocrine abnormalities associated with APS1, and clinicians should be cognizant of the association of treatable anemia in patients with APS1. PMID- 14755374 TI - Intralobar pulmonary sequestration in a patient with non-Hodgkin's lymphoma: a cause of confusion. AB - Pulmonary sequestration, a rare congenital pulmonary disorder, is characterized by nonfunctioning lung tissue that is separated from normal tracheobronchial tree. We present a 60-year-old woman with diffuse large cell non-Hodgkin's lymphoma. After 6 cycles of chemotherapy, paratracheal and aorticopulmonary lymphadenopathies had disappeared. However, the size of the pulmonary mass in the left lower lobe had persisted. Percutaneous fine-needle aspiration biopsy of the pulmonary mass was not diagnostic, so thoracotomy was applied. The lesion was defined as pulmonary sequestration, and basal segmentectomy was performed. After proper and sufficient chemotherapy, histopathological diagnosis of any persisting masses should be confirmed prior to overtreatment decision. PMID- 14755375 TI - Chronic myeloid leukemia with an e13a3 BCR-ABL fusion: benign course responsive to imatinib with an RT-PCR advisory. AB - A case of a patient with chronic myeloid leukemia whose cells expressed an e13a3 (b2a3) variant BCR-ABL p210 mRNA is presented. The variant splice was detected by a qualitative reverse transcriptase polymerase chain reaction using primers complementary to BCR exon 13 (b2) and ABL exon 3 (a3). The patient responded well to imatinib and achieved a complete cytogenetic response. PMID- 14755376 TI - Clinical usefulness of a functional assay for the von Willebrand factor cleaving protease (ADAMTS 13) and its inhibitor in a patient with thrombotic thrombocytopenic purpura. AB - Decreased von Willebrand factor cleaving protease activity (VWFCP, ADAMTS 13) leads to persistence of unusually large multimers of von Willebrand factor that bind to platelets, causing platelet aggregates, microangiopathic hemolysis, and thrombocytopenia in patients with thrombotic thrombocytopenic purpura (TTP). The clinical value of measuring ADAMTS 13 and its inhibitor is not fully defined; the case reported here illustrates the usefulness of the assay to help confirm the clinical diagnosis in a patient with other potential causes for thrombotic microangiopathy; the assay also helped in making treatment decisions. A patient with systemic lupus erythematosis (SLE) presented with fever and abdominal pain, thrombocytopenia, and anemia. Thrombotic microangiopathy was diagnosed by the appearance of schistocytes, decreasing platelet count, and evidence of hemolysis. ADAMTS 13 was decreased and an inhibitor was demonstrated in the patient's initial blood sample within 24 hr of admission. Plasma exchange was initiated, and serial assays showed increased ADAMTS 13 activity and decreased inhibitor after each plasma exchange; there was a rebound in inhibitor and a decrease in ADAMTS 13 activity prior to the next exchange that lessened over time. Increasing levels of protease activity correlated with clinical and laboratory improvement. Measurement of ADAMTS 13 activity and its inhibitor aided in the diagnosis of this complicated case of a patient with other potential causes for microangiopathic hemolysis. Subsequent levels correlated with the clinical course, and disappearance of the inhibitor indicated that long-term plasma exchange or other immunosuppressive treatment was not needed. PMID- 14755377 TI - Serum level of interleukin-16 in multiple myeloma patients and its relationship to disease activity. AB - Interleukin-16 (IL-16) is a chemoattractant of CD4+ lymphocytes, and it has been implicated in the pathogenesis of various inflammatory diseases. There is evidence that it may have a role in multiple myeloma (MM). In the present study, we determined the serum level of IL-16 both before and after treatment of MM and related it to inflammatory markers and survival. Forty-eight newly diagnosed MM patients were included in the study. Disease stage was defined using the Durie Salmon classification system (10 patients were in stage I, 19 in stage II, and 19 in stage III). After standard treatment, 22 patients reached the plateau phase and were re-evaluated. The following serum parameters were measured: IL-16, IL-6, alpha-1 antitrypsin (alpha1AT), and C-reactive protein (CRP). Survival was determined as the number of months elapsed since original diagnosis. The mean +/- SD of serum IL-16 was 343 +/- 195 pg/ml in the pre-treatment MM group and 101 +/- 30 pg/ml in the control group. All measured parameters were higher in the patient group compared to healthy controls. Furthermore, IL-16, IL-6, alpha1AT, and CRP were significantly increased with increasing stage of disease, from stage I to stage III (P<0.01). All parameters decreased significantly following effective chemotherapy (P<0.002). Patients with a high level of IL-16 (>430 pg/ml) displayed an inferior survival time in comparison to those with lower levels of IL-16. In the pre-treatment group, IL-16 correlated with alpha1AT and IL-6 (r=0.374, P<0.01 and r=0.454, P<0.002, respectively). IL-16 may play a role in multiple myeloma; however, further functional studies are required. PMID- 14755378 TI - Simultaneous manifestation of chronic lymphocytic leukemia (CLL) and hairy cell leukemia (HCL). AB - We report a unique case of 83-year-old Caucasian male with the initial simultaneous manifestation of chronic lymphocytic leukemia (CLL) and hairy cell leukemia (HCL). The patient presented with absolute lymphocytosis in the blood, asymptomatic generalized lymphadenopathy, and mild splenomegaly. The diagnosis of CLL was suggested from the blood film, but subsequent flow cytometric (FC) analysis on peripheral blood mononuclear cells (PBMNC) revealed two distinct abnormal clones of mature B cells. A small subpopulation (7%) of lymphoid cells expressed CD20, CD11c, FMC-7, CD103, CD25, and kappa surface light chain, consistent with HCL. The larger subpopulation (75%) of lymphoid cells expressed CD19, CD20, CD23, CD5, and lambda light chain, consistent with CLL. The expression of different immunoglobulin light chains on the circulating CLL (lambda) and HCL (kappa) cells suggested two, independent, malignant B-cell clones. Interestingly, FC analysis of bone marrow (BM) cells done 6 months later revealed bright lambda light chain expression on the HCL cells. Despite administration of several different courses of chemotherapy, the HCL subpopulation was not eliminated from the BM but remained stable between 7% and 10% of total BM lymphoid cells. The CLL, responsible for most of clinical symptoms in our patient, responded to combination chemotherapy with fludarabine and cytoxan, and later to monotherapy with rituximab. PMID- 14755379 TI - Unexplained bone marrow granulomas: is amiodarone the culprit? A report of 2 cases. AB - Granulomas in the bone marrow are usually caused by infectious or hematological diseases, and drugs are only rarely implicated as causative agents. Recent reports have drawn attention to the role of amiodarone in the etiology of bone marrow granulomas. We report two cases of amiodarone-induced bone marrow granulomas in patients being investigated for refractory anemia and pancytopenia, respectively. Since both patients had life-threatening arrhythmias, discontinuation of the drug followed by rechallenge was not possible. Both patients did well in spite of continued amiodarone therapy, indicating that the underlying hematological illnesses were unrelated to the granulomas. Amiodarone should be considered as a possible cause of bone marrow granulomas after the exclusion of other causes. Continued use of amiodarone after granuloma formation must be dictated by the underlying cardiac condition. PMID- 14755380 TI - Hepatosplenic gammadelta T-cell lymphoma in a 10-year-old boy successfully treated with hematopoietic stem cell transplantation. AB - The authors report a 10-year-old boy with hepatosplenic gammadelta T-cell lymphoma, a rare form of lymphoma that is highly aggressive, exceedingly rare in children, and primarily seen in young men. Conventional multi-agent chemotherapy appears to be inadequate for cure. This is the first report with this type of lymphoma in a boy less than 15 years old treated with hematopoietic stem cell transplantation (HSCT). PMID- 14755381 TI - Long-standing Bell's palsy and ipsilateral conjunctival maltoma. PMID- 14755382 TI - Coagulopathy in a patient with X-linked hyper-IgM syndrome who developed Kaposi's sarcoma. PMID- 14755383 TI - Long-term outcome of splenectomy for immune thrombocytopenic purpura. PMID- 14755384 TI - Thalassemia beta0 due to an identical frameshift mutation, codon 15 (-T) in both parents. PMID- 14755385 TI - Pancytopenia due to extensive hemophagocytosis following anti-tubercular treatment. PMID- 14755386 TI - Immune thrombocytopenia in a newborn from a mother with primary antiphospholipid syndrome. PMID- 14755387 TI - Electronic monitoring of variation in drug intakes can reduce bias and improve precision in pharmacokinetic/pharmacodynamic population studies. AB - Population pharmacokinetic (PK) and pharmacodynamic (PD) studies evaluate drug concentration profiles and pharmacological effects over time when standard drug dosage regimens are assigned. They constitute a scientific basis for the determination of the optimal dosage of a new drug. Population PK/PD analyses can be performed on relatively few measures per patient enabling the study of a sizable sample of patients who take the drug over a possibly long period of time. We expose the problem of bias in PK/PD estimators in the presence of partial compliance with assigned treatment as it occurs in practice. We propose to solve this by recording accurate data on a number of previous dose timings and using timing-explicit hierarchical non-linear models for analysis. In practice, we rely on electronic measures of an ambulatory patient's drug dosing histories. Especially for non-linear PD estimation, we found that not only bias can be reduced, but higher precision can also be retrieved from the same number of data points when irregular drug intake times occur in well-controlled studies. We apply methods proposed by Mentre et al. to investigate the information matrix for hierarchical non-linear models. This confirms that a substantial gain in precision can be expected due to irregular drug intakes. Intuitively, this is explained by the fact that regular takers experience a relatively small range of concentrations, which makes it hard to estimate any deviation from linearity in the effect model. We conclude that estimators of PK/PD parameters can benefit greatly from information that enters through greater variation in the drug exposure process. PMID- 14755388 TI - Testing non-inferiority (and equivalence) between two diagnostic procedures in paired-sample ordinal data. AB - Before adopting a new diagnostic procedure, which is more convenient and less expensive than the standard existing procedure, it is essentially important to assess whether the diagnostic accuracy of the new procedure is non-inferior (or equivalent) to that of the standard procedure. In this paper, we consider the situation where test responses are on an ordinal scale with more than two categories. We give two definitions of non-inferiority, one in terms of the probability of correctly identifying the case for a randomly selected pair of a case and a non-case over all possible cut-off points, and the other in terms of both the sensitivity and specificity directly. On the basis of large sample theory, we develop two simple test procedures for detecting non-inferiority. We further conduct Monte Carlo simulation to evaluate the finite sample performance of these test procedures. We note that the two asymptotic test procedures proposed here can actually perform reasonably well in a variety of situations even when the numbers of studied subjects from the diseased and non-diseased populations are not large. To illustrate the use of the proposed test procedures, we include an example of determining whether the diagnostic accuracy of using a digitized film is non-inferior to that of using a plain film for screening breast cancer. Finally, we note that the extension of these results to accommodate the case of detecting (two-sided) equivalence is simply straightforward. PMID- 14755389 TI - Admissible two-stage designs for phase II cancer clinical trials. AB - In a typical two-stage design for a phase II cancer clinical trial for efficacy screening of cytotoxic agents, a fixed number of patients are initially enrolled and treated. The trial may be terminated for lack of efficacy if the observed number of tumour responses after the first stage is too small, thus avoiding treatment of patient with inefficacious regimen. Otherwise, an additional fixed number of patients are enrolled and treated to accumulate additional information on efficacy as well as safety. The minimax and the so-called 'optimal' designs by Simon have been widely used, and other designs have largely been ignored in the past for such two-stage cancer clinical trials. Recently Jung et al. proposed a graphical method to search for compromise designs with features more favourable than either the minimax or the optimal design. In this paper, we develop a family of two-stage designs that are admissible according to a Bayesian decision theoretic criterion based on an ethically justifiable loss function. We show that the admissible designs include as special cases the Simon's minimax and the optimal designs as well as the compromise designs introduced by Jung et al. We also present a Java program to search for admissible designs that are compromises between the minimax and the optimal designs. PMID- 14755390 TI - Parametric randomization-based methods for correcting for treatment changes in the assessment of the causal effect of treatment. AB - We develop parametric maximum likelihood methods to adjust for treatment changes during follow-up in order to assess the causal effect of treatment in clinical trials with time-to-event outcomes. The accelerated failure time model of Robins and Tsiatis relates each observed event time to the underlying event time that would have been observed if the control treatment had been given throughout the trial. We introduce a bivariate parametric frailty model for time to treatment change and time to trial endpoint. Estimating equations which respect the randomization are constructed and compared to maximum likelihood methods in a simulation study. The Concorde trial of immediate versus deferred zidovudine in HIV infection is used as a motivating example and illustration of the methods. PMID- 14755391 TI - Two-sample scale tests for comparison of metabolic rates for styrene in previously exposed and unexposed groups. AB - Statistical analysis procedure in two-sample treatment difference, as well as alternative hypothesis, may play a central role in statistical inference, especially in small sample size case. In this paper, simple non-parametric two sample permutation tests for scale difference based on the Hodges-Lehmann estimator are reformulated, and are applied to a study examining whether previous styrene exposure increases (thus a one-sided alternative hypothesis rather than a two-sided one) the human liver's metabolic ability to convert styrene into styrene oxide, in which a three-compartment physiologically based pharmacokinetic (PBPK) model was used to compare the estimated metabolic constant kappaamong the previously exposed and unexposed groups. Contrary to the previous conclusion, the proposed tests for scale difference identified from a mixed-effects model showed a significant result. PMID- 14755392 TI - A Bayesian analysis of the 4-year follow-up data of the Wisconsin epidemiologic study of diabetic retinopathy. AB - The Wisconsin epidemiologic study of diabetic retinopathy (WESDR) is a population based epidemiologic study carried out in Southern Wisconsin during the 1980s of the last century. The resulting data were analysed by different statisticians and ophthalmologists during the last two decades. Most of the analyses were carried out on the baseline data, although there were two follow-up studies on the same population. In this present paper, we provide a Bayesian analysis of the first follow-up data which were taken 4 years after the baseline study. Our Bayesian analysis provides estimates of the associated covariate effects. Choice of the best model in terms of the covariate inclusion is also performed. The baseline data were used to set the prior for the parameters. Extensive numerical computations illustrate our present methodology. PMID- 14755393 TI - Analysis of testicular cancer data using a frailty model with familial dependence. AB - Previously published papers have indicated a fairly strong familial dependence intesticular cancer patients. This is particularly evident in brothers. We have applied a frailty model with familial dependence to family data on brothers of testicular cancer patients from the Norwegian Radium Hospital. The model is a two level frailty, with variation in susceptibility at both the family and the individual level. Specifically, the frailty variable is assumed to be compound Poisson distributed to allow individuals to be non-susceptible. The underlying Poisson parameter is gamma distributed to model how testicular cancer is distributed among families. This is an extension of a previous compound Poisson frailty model developed for individual testicular cancer data, and an alternative to traditional modelling of survival time family data. The likelihood construction and ascertainment problems are looked at in detail. To avoid ascertainment bias, the likelihood is based on the probability of observing the disease status for each brother in a family, given that at least one brother is ascertained. The estimated relative risk for brothers is 7.4. This paper expands on a previous analysis of the data by using a frailty model, which makes it possible to examine how the cancer is distributed among families. The estimated gamma-shaped parameter is 0.151 (95 per cent confidence interval 0.078-0.294), and this indicates that in order to obtain the high relative risks observed for brothers of testicular cancer patients, the distribution of susceptibility has to be strongly skewed among the families. The vast majority of families have a very low risk and a small proportion have a high risk. In addition, a quantity similar to the relative risk is derived to show that the susceptibility is skewly distributed also if the Poisson parameter is Bernoulli or stable distributed. This indicates that the results are valid also if other distributions are used to model familial dependence in the compound Poisson frailty model. PMID- 14755394 TI - Stochastic model for non-standard case-cohort design. AB - Application of case-cohort design to multi-state disease progression in epidemiological studies has been barely addressed. To estimate multi-state disease natural history, we proposed non-homogeneous exponential regression stochastic model to accommodate the data requiring a non-standard case-cohort design. We allowed transition rates to vary with time by modelling the time of transitions between two states with Weibull distribution. The exponential regression model was used to assess the effect of patient-specific covariates on multi-state disease progressions. This method was successfully applied to two epidemiological applications. The first application was to elucidate the effect of betel quids, smoking and alcohol on three-state disease progression, from normal, through leukoplakia and finally to oral cancer. The second application was to extend the three-state to a five-state model to estimate transition rates from normal to diminutive adenoma to small adenoma to large adenoma and finally to invasive carcinoma of the colon and rectum. Finally, an index for assessing the treatment efficacy for pre-cancerous lesion was developed by comparing transition probabilities derived from the proposed model with the probabilities of malignant transformation after a medical regime. PMID- 14755395 TI - Adaptation to life after surgical removal of the bladder-an application of graphical Markov models for analysing longitudinal data. AB - Graphical Markov models have been developed particularly for the analysis of observational data. They allow the control of various background variables when analysing theoretically relevant associations. This paper demonstrates the application and some advantages of graphical Markov models in comparison to conventional statistical analyses. The aim of the study was to identify patients at risk for developing decreased health-related quality of life (QoL) after cystectomy and to explore the influence of coping on QoL in this situation. Therefore, the method was applied to analyse the data of a prospective study, in which 81 patients with bladder cancer were interviewed pre-operatively and in a 1 year follow-up. QoL was assessed both times, and two basic coping strategies (active and depressive) were measured preoperatively. The explanatory variables of theoretical interest were active and depressive coping strategies. As a result of the analysis, relevant proportions of variance in the development of QoL could be explained by the suggested model (60 per cent in mental component, 40 per cent in physical component of QoL). Active coping was positively related to QoL, depressive coping negatively. These effects were linear in the physical component of QoL, moderated by working status and the type of urinary diversion in the mental component of QoL. PMID- 14755396 TI - Binary latent variable modelling and its applicationin the study of air pollution in Hong Kong. AB - A binary latent variable is constructed to account for the correlation between multiple binary outcomes and is treated as a dependent variable in modelling for covariate effects. This modelling method is similar to the structural equation modelling. Three models are considered: (1) all covariates affecting the binary latent variable directly; (2) some covariates affecting the binary latent variable while other affecting the manifestation of the binary outcomes; and (3) no covariates are included. Gibbs sampling, a special case of the Markov Chain Monte Carlo method, is used to estimate the parameters in the models. Simulation studies show that this method is valid and reliable in estimating covariate effects. But Model (1) fitted the data best with lowest value in the deviance information criteria. The method is illustrated by applying it to the data analysis of an environmental air pollution study. The results show that air pollution (i.e. the most versus the least polluted district) (odds ratio 1.20; 95% confidence interval 0.97-1.49; p=0.102), smoking (relative to not smoking) (2.75; 2.21-3.41; p < 0.001) and mosquito coil use (relative to non-use) (1.27; 0.99-1.62; p=0.058) had an impact on the respiratory health of male adults in Hong Kong. PMID- 14755397 TI - Comment on: confidence limits for the ratio of two rates based on likelihood scores: non-iterative method. PMID- 14755399 TI - "Confidence limits for the ratio of two rates based on likelihood scores: non iterative method," by Graham PL et al. PMID- 14755401 TI - Multi-channel magnetocardiography for detecting beat morphology variations in fetal arrhythmias. AB - OBJECTIVE: Over the last few years, a number of studies have shown that fetal magnetocardiography (fMCG) is useful in describing fetal cardiac activity. A 55 channel MCG system in Ulm was used to record fetal cardiac activity in 12 pregnant women (with normal fetal heart activity in echocardiography) and in 5 pregnant women in whom the echocardiography showed fetal arrhythmias. METHOD: The recorded MCG data were treated in order to eliminate the maternal signal and three MCG channels with the best signals were used to emulate a standard electrocardiogram (ECG) recording so that standard MCG analysis could be performed. RESULTS: The results in assessing fetal electrophysiology, demonstrating its potential, are presented here for two fetuses with recorded supraventricular extrasystoles (SVES) and for one with ventricular extrasystoles (VES). Concerning the SVES, the analysis software OMEGA was able to separate three different beat morphologies. The VES could be detected exactly and have been confirmed by postnatal ECG. CONCLUSION: The beat morphology and the beat-to beat variations allow new insights into the electrophysiology of the fetal heart. PMID- 14755402 TI - The prenatal diagnosis of cloacal dysgenesis sequence in six cases: can the termination of pregnancy always be the first choice? AB - OBJECTIVE: Cloacal dysgenesis sequence is a lethal malformation, which usually requires termination. In this study, our aim was to evaluate the prenatal and postnatal diagnostic features of cloacal dysgenesis sequence and review the management of the patients. MATERIAL AND METHODS: The data of six cases of cloacal dysgenesis sequences were collected from the ultrasonography and neonatal records. The findings were evaluated in prenatal and postnatal periods. Chromosomal analysis was performed in all the cases. The evaluation of primary and secondary malformations was done. Coexisting anomalies were searched for by radiology and histopathology. RESULTS: Malformations in six cases (two females and four males) were described. The absence of anal, genital, and urinary openings with intact perineum covered by smooth skin were common findings. These features were considered as primary malformations for cloacal dysgenesis sequence. Secondary anomalies (urinary and gastrointestinal system malformations, pulmonary hypoplasia, and other coexisting anomalies) were evaluated. CONCLUSION: The prenatal differential diagnosis of cloacal dysgenesis sequence from other urinary obstructive diseases was essential regarding fetal prognosis, prenatal, and neonatal management. The bladder outlet obstruction and pulmonary hypoplasia due to reduced amniotic fluid and/or kidney disease were considered prognostic factors for neonatal death. Termination of pregnancy is almost always recommended instead of intrauterine shunt procedures; but if we take into consideration one of our cases and a few reported cases who survived in the neonatal period, the prenatal management of these pregnancies needs to be reevaluated. PMID- 14755403 TI - Middle cerebral artery Doppler velocimetric assessment in two cases of hydrops fetalis without fetal anaemia. AB - OBJECTIVES: Raised middle cerebral artery (MCA) Doppler velocity has been shown to be highly predictive of moderate to severe fetal anaemia. We present two false positive cases of raised MCA Doppler velocity in non-immune hydropic fetuses. METHODS: In both cases, routine investigations for fetal hydrops, as well as detailed ultrasound scanning and MCA peak-systolic velocity Doppler assessment (maximum velocity taken at zero degrees to the vessel), were performed. Fetal blood sampling was carried out at the same visit. RESULTS: MCA peak-systolic velocity values greater than 1.50 MoM for gestation were found in both cases. However, both fetuses had normal haemoglobin values and haematocrits. Both fetuses died in utero soon after diagnosis. CONCLUSIONS: A raised MCA velocity in a hydropic fetus may not always be due to fetal anaemia, rather it may indicate a fetus in poor condition perhaps due to cardiovascular decompensation and redistribution of blood to the fetal brain. PMID- 14755404 TI - Prenatal diagnosis of premature centromere division-related mosaic variegated aneuploidy. AB - OBJECTIVES: To present the prenatal diagnosis of premature centromere division (PCD)-related mosaic variegated aneuploidy (MVA) and a review of the literature. CASE AND METHODS: A 33-year-old primigravida woman underwent amniocentesis at 22 weeks' gestation because of intrauterine growth restriction (IUGR), microcephaly, and oligohydramnios. Amniocentesis revealed PCD-related MVA. Repeat amniocentesis two weeks later consistently showed PCD-related MVA. The pregnancy was terminated. The proband postnatally manifested dysmorphic facial features of microcephaly, hypertelorism, low-set ears, a broad nasal bridge, a thin upper lip, and overriding toes. At autopsy, the internal organs were unremarkable. Cytogenetic analyses of the cord blood, liver, lungs, skin, and placenta displayed PCD-related MVA in all tissues studied. The PCD frequencies for the cells in the amniotic fluid (first culture), amniotic fluid (second culture), cord blood, liver, lungs, skin, and placenta were 53.3%, 56.5%, 47.4%, 38.7%, 33.9%, 33.3%, and 40.8% respectively. CONCLUSION: The present case provides evidence that, in cases of pregnancy with PCD-related MVA, the cytogenetic result of the amniocytes correlates well with those of the fetal cells and chorionic villi cells. We suggest that prenatal sonographic detection of a complex of IUGR, microcephaly and oligohydramnios with or without central nervous system abnormalities should include a differential diagnosis of PCD-related MVA. PMID- 14755405 TI - Prenatal sonographic diagnosis of congenital hiatal hernia. AB - OBJECTIVES: To report a rare case of congenital hiatal hernia illustrating the importance of its prenatal diagnosis as well as to discuss the prenatal sonographic criteria. CASE REPORT: A case of congenital hiatal hernia was diagnosed by ultrasound at 33 weeks of gestation. After a normal second-trimester morphologic ultrasound examination, a hypoechogenic mass was detected in the posterior mediastinum juxtaposed to the vertebral body and seemed to be in continuity with the intra-abdominal stomach bubble. Congenital hiatal hernia was suspected mainly because of the dynamic position of the stomach during the examination, without mediastinal shift, and normal appearance of the diaphragm on parasagittal sections of the thorax. Postnatal management was planned with no urgency and surgery was successfully performed, confirming the diagnosis. CONCLUSION: This rare case illustrates the importance of prenatal diagnosis of congenital hiatal hernia for prenatal counseling and postnatal management. The ultrasound criterion for prenatal diagnosis is the presence of a herniated stomach in the posterior mediastinum, sometimes having a dynamic position during examination, with no mediastinal shift associated with normal diaphragm appearance on parasagittal sections of the thorax. PMID- 14755406 TI - Second-trimester diagnosis of intracranial vascular anomalies in a fetus with subdural hemorrhage. AB - OBJECTIVES: Risk factors for intracranial hemorrhage occurring in prenatal life are imperfectly known. A case of prenatal diagnosis of subdural hemorrhage associated with multiple intracranial vascular aneurysms is described. METHODS: Sonography and magnetic resonance imaging of the fetal head were obtained at 21 weeks' gestation and compared with pathologic findings. RESULTS: Sonography showed a large transonic mass displacing the normal intracranial structures. Magnetic resonance imaging demonstrated the hemorrhagic origin of the mass and showed multiple vascular anomalies. Postmortem examination confirmed the compression of the cerebral hemisphere by a blood collection, probably because of bleeding from one of the multiple vascular aneurysms into the subdural space. CONCLUSION: Magnetic resonance imaging with the use of single-shot ultrafast sequences may be useful not only in the differential diagnosis of fetal intracranial hemorrhage but also in identifying vascular risk factors. PMID- 14755407 TI - Follow-up of children with isolated fetal echogenic bowel with particular reference to bowel-related symptoms. AB - OBJECTIVES: To determine whether there was any evidence of long-term bowel pathology in children, apparently healthy at birth, who had a prenatal second trimester diagnosis of isolated grade 2 fetal echogenic bowel. METHODS: This was a retrospective study using data from the Oxford Congenital Anomaly Register. Fetuses with isolated grade 2 fetal echogenic bowel and date of delivery from 1994 to 2000 inclusive were identified. Information about the health of the children, particularly relating to bowel symptoms, was obtained from hospital records and from a questionnaire sent to the general practitioner. RESULTS: A total of 109 cases were identified, with delivery details available for 108. There was one unexplained intrauterine death, and additional problems were subsequently diagnosed in four cases (cystic fibrosis (2), Down syndrome (1), and VACTERL (1)). Questionnaires were sent to the GPs of the 103 who had no problems identified at the time of discharge from the maternity hospital. Age at follow-up ranged from one to four years. Responses to the questionnaires were received from 83 (81%). Of these, 74 (89%) had not reported bowel symptoms to the GP, 9 (11%) reported symptoms relating to constipation (6), chronic abdominal pain (1), infantile colic with milk intolerance (1) and gastro-oesophageal reflux (1). CONCLUSION: This small study provides some reassurance that there was no evidence of any serious long-term bowel pathology associated with isolated fetal echogenic bowel. PMID- 14755408 TI - Clinical, cytogenetic, and molecular findings of prenatally diagnosed mosaic trisomy 4. AB - OBJECTIVES: To present the clinical, cytogenetic, and molecular findings of prenatally diagnosed mosaic trisomy 4. CASE: An amniocentesis was performed at 21 weeks' gestation because of maternal anxiety. Cytogenetic analysis revealed mosaicism for trisomy 4, 47,XX,+4[4]/46,XX[16]. Level II ultrasound demonstrated tetralogy of Fallot. Repeated amniocentesis at 23 weeks' gestation revealed 47,XX,+4[4]/46,XX[19]. The pregnancy was terminated. Phenotypic findings included tetralogy of Fallot, hypertelorism, micrognathia, abnormal ears, duplicated phalanges of the left thumb, clinodactyly, and overlapping of the toes. The karyotype of the cord blood was 46,XX. Cytogenetic analyses of the multiple tissue samplings showed a karyotype of 47,XX,+4 in 40/40 cells of the amniotic membrane (amnion), and 47,XX,+4/46,XX with various levels of trisomy 4 in the cells of the liver, lungs, placenta, skin, and umbilical cord. The levels of trisomy 4 were 11/40 in the liver, 8/40 in the lungs, 31/40 in the placenta, 9/40 in the skin, and 8/40 in the umbilical cord. METHOD: The parental origin and meiotic origin of trisomy 4 were determined by examining the amniotic membrane using quantitative fluorescent polymerase chain reaction assays with polymorphic markers specific for chromosome 4. The result was consistent with a paternal meiosis I nondisjunction error. The cord blood showed a biparental inheritance. An extra paternal heterozygous allele with partial dosage increase was noted in other fetal and extraembryonic tissues studied. CONCLUSION: A diagnosis of trisomy 4 mosaicism in amniocytes indicates an increased risk for fetal abnormalities. Associated abnormal findings include congenital heart defects and anomalies of the digits and thumb. A confirmatory placental sampling may be helpful, whereas a fetal blood sampling is of a very limited value. A postnatal amnion sampling may provide additional clues to the fetal involvement of trisomy 4. PMID- 14755409 TI - Radiographic and genetic diagnosis of sporadic hypochondroplasia early in the neonatal period. AB - Hypochondroplasia is an autosomal dominant skeletal dysplasia expressing postnatal onset of short stature with mild rhizomelic shortening of the limbs. This manifestation leads to restricted prenatal diagnosis of the disorder. We report here on a sporadic case of a hypochondroplastic baby, whose prenatal sonographic measurements were serially recorded from 19 weeks of gestation. Mild shortening of the limbs became manifest after 26 weeks of gestation. Biparietal diameter was within the normal range throughout gestation. Both parents were of average stature. A tentative diagnosis of a nonlethal short-limb skeletal dysplasia was made. At birth, the clinical manifestations of the neonate were not characteristic, but the radiographic features raised the possibility of hypochondroplasia. Molecular analyses revealed a C to G mutation at nucleotide 1659 of the fibroblast growth factor receptor 3 (FGFR3) gene, a common mutation in hypochondroplasia. PMID- 14755410 TI - Prenatal diagnosis of mosaic distal 5p deletion and review of the literature. AB - OBJECTIVES: To present the prenatal diagnosis of mosaic distal 5p deletion and a review of the literature. CLINICAL SUBJECT AND METHODS: A 37-year-old woman, gravida 2, para 1, underwent genetic amniocentesis at 17 weeks' gestation because of advanced maternal age. Cytogenetic analysis of the cultured amniocytes revealed mosaicism for a distal 5p deletion, mos 46,XX,del(5)(p15.1)/46,XX (23 colonies/23 colonies). Repeat amniocentesis showed a consistent karyotype of mos 46,XX,del(5)(p15.1)/46,XX (12 colonies/15 colonies). The parental karyotypes were normal. Prenatal ultrasound demonstrated microcephaly and cerebellar hypoplasia. The pregnancy was terminated at 21 weeks' gestation. Postnatally, the fetus displayed microcephaly, a triangular face, hypertelorism, epicanthic folds, down slanting palpebral fissures, low-set ears, and micrognathia. A karyotype of mos 46,XX,del(5)(p15.1)/46,XX was found in the cord blood, liver, lungs, and skin, whereas the placenta had a different karyotype of mos 46,XX,dup(5)(qter- >p15.3::p15.3-->p10)/46,XX, and the karyotype of the amnion was mos 46,XX,del(5)(p15.1)/46,XX,dup(5)(qter-->p15.3::p15.3-->p10)/46,XX,trp(5)(qter- >p15.3::p15.3-->p10::p10-->p15.3)/46,XX. The deletion, duplication, and triplication of the terminal region of the short arm of chromosome 5 were confirmed by the studies of fluorescence in situ hybridization. CONCLUSION: The cri-du-chat syndrome can be identified prenatally because of advanced maternal age, familial cri-du-chat syndrome, parental balanced translocations involving chromosome 5, sonographically detected fetal structural abnormalities, and/or an abnormal maternal serum test. Fetuses with the mosaic distal 5p deletion may be associated with the sonographic findings of microcephaly and cerebellar hypoplasia, and fetoplacental and fetoamnionic chromosomal discrepancies. PMID- 14755411 TI - Interleukin-8 mRNA in vaginal secretions: a prenatal marker of congenital infection in case of preterm labor with intact membranes. AB - OBJECTIVE: To study whether interleukin-8 (IL-8) mRNA in vaginal secretions is associated with congenital infection and preterm delivery in the case of preterm labor with intact membranes. METHODS: This prospective clinical study in a tertiary referral center included 280 patients who gave birth to 360 infants from 1997 through 1999. IL-8 mRNA in vaginal secretions was determined with reverse transcriptase polymerase chain reaction. Logistic regression was used to examine the association between vaginal IL-8 mRNA and congenital infection independently of the time of birth. Main outcome measures were congenital infection and delivery before 37 and 33 weeks' gestation. RESULTS: A total of 100 women (100/280 (35.7%)) gave birth before 37 weeks. A total of 54 children (54/360 (15%)) had congenital infection. IL-8 mRNA in vaginal secretions was associated with delivery within 14 days of the sampling (24 (15.6%) vs. 7 (5.6%) p < 0.01), but not with delivery within 48 h, 7 days (p = 0.07) or before 37 or 33 weeks. There were more congenital infections in the group with detectable IL-8 mRNA (37 (19.3%) than in the negative group (17 (10.1%); p < 0.05). IL-8 mRNA was associated with congenital infection independently of the time of birth (OR: 2.6 (1.3-5.1)). This test had a sensitivity for predicting neonatal infection of 69%. Its specificity was 49%, its positive predictive value 19%, and its negative predictive value 90%. CONCLUSION: IL-8 mRNA could be a prenatal noninvasive vaginal marker of congenital infection. PMID- 14755412 TI - Prenatal diagnosis of mosaic trisomy 16 associated with congenital diaphragmatic hernia and elevated maternal serum alpha-fetoprotein and human chorionic gonadotrophin. AB - OBJECTIVES: To present the clinical, cytogenetic, and molecular findings of prenatally diagnosed mosaic trisomy 16. CASE: A 30-year-old gravida 2, para 1 woman was referred for amniocentesis because of a positive maternal serum screen result with elevated maternal serum alpha-fetoprotein (MSAFP) and maternal serum free beta-human chorionic gonadotrophin (MSfreebeta-hCG). Cytogenetic analysis of amniotic fluid at 21 weeks' gestation revealed mosaicism for trisomy 16, 47,XX,+16[3]/46,XX[15]. Ultrasonography demonstrated right diaphragmatic hernia and agenesis of left umbilical artery. The pregnancy was terminated subsequently. The karyotype of the cord blood was 46,XX. Cytogenetic analyses of the multiple sampled tissue specimens showed a karyotype of 47,XX,+16 in the placenta and 47,XX,+16/46,XX with various levels of trisomy 16 in the umbilical cord and skin. Molecular studies showed that the trisomy 16 in the placenta was likely to have resulted from a maternal meiosis II nondisjunction error. Partial dosage increase of an extra maternal allele was noted in the skin and umbilical cord. CONCLUSION: Fetuses with mosaic trisomy 16 may be associated with congenital diaphragmatic hernia and elevated MSAFP and MShCG. Fetal blood sampling is of a limited value in confirming mosaic trisomy 16 ascertained through amniocentesis. PMID- 14755413 TI - Screening approach for Fragile X syndrome. PMID- 14755415 TI - Prospective prenatal diagnosis of Fraser syndrome variant in a family with negative history. PMID- 14755416 TI - Coping with termination of pregnancy for fetal abnormality. PMID- 14755419 TI - Hybrid germanium/silica optical fibers for endoscopic delivery of erbium:YAG laser radiation. AB - BACKGROUND AND OBJECTIVES: Endoscopic applications of the erbium (Er):YAG laser have been limited due to the lack of an optical fiber delivery system that is robust, flexible, and biocompatible. This study reports the testing of a hybrid germanium/silica fiber capable of delivering Er:YAG laser radiation through a flexible endoscope. STUDY DESIGN/MATERIALS AND METHODS: Hybrid optical fibers were assembled from 1-cm length, 550-microm core, silica fiber tips attached to either 350- or 425-microm germanium oxide "trunk" fibers. Er:YAG laser radiation (lambda = 2.94 microm) with laser pulse lengths of 70 and 220 microseconds, pulse repetition rates of 3-10 Hz, and laser output energies of up to 300 mJ was delivered through the fibers for testing. RESULTS: Maximum fiber output energies measured 180+/-30 and 82+/-20 mJ (n = 10) under straight and tight bending configurations, respectively, before fiber interface damage occurred. By comparison, the damage threshold for the germanium fibers without silica tips during contact soft tissue ablation was only 9 mJ (n = 3). Studies using the hybrid fibers for lithotripsy also resulted in fiber damage thresholds (55-114 mJ) above the stone ablation threshold (15-23 mJ). CONCLUSIONS: Hybrid germanium/silica fibers represent a robust, flexible, and biocompatible method of delivering Er:YAG laser radiation during contact soft tissue ablation. However, significant improvement in the hybrid fibers will be necessary before they can be used for efficient Er:YAG laser lithotripsy. PMID- 14755418 TI - Photodynamic endometrial ablation for the treatment of dysfunctional uterine bleeding: a preliminary report. AB - BACKGROUND AND OBJECTIVES: To evaluate feasibility and functional effects of photodynamic endometrial ablation (PEA) in patients. STUDY DESIGN/PATIENTS AND METHODS: A total of 15 PEAs has been performed in 11 patients using topically applied 5-aminolevulinic acid (ALA) solutions and a balloon-light diffuser (160 J/cm(2), 635 nm). Uterine bleeding intensity has been determined on a daily basis 3 months prior to and up to 6 months after endometrial ablation using an analogous scale scoring from 1 (spotting) to 6 U (severe bleeding). Statistical analysis by unpaired Student's t-test. RESULTS: The mean number of bleeding units per cycle (n = 44) was 35.7 prior to PEA. The decrease in bleeding units was significant for the months 1-3 (24.4 U per cycle; P = 0.03), but not for the months 4-6 (25.9 U; P = 0.11) following PEA. CONCLUSIONS: PEA is feasible and provides a significant short-term reduction of uterine bleeding. PMID- 14755420 TI - Effect of the combination of laser excision and interstitial hyperthermia in palliative therapy of head and neck tumours in the advanced stage of the disease. AB - BACKGROUND AND OBJECTIVES: We evaluated the potential of combining laser excision with laser interstitial hyperthermia in order to improve the quality of life of patients with 3rd or 4th stage malignant head and neck tumours. STUDY DESIGN/MATERIALS AND METHODS: To evaluate the quality of life, we used the 4th version of the University of Washington Quality of Life assessment score modified for head and neck tumours. The study group of 20 patients assessed their condition in a questionnaire, examining 12 aspects of their quality of life before the laser treatment, the second day after, and 6 weeks after the treatment. RESULTS: We observed a gain of 16.41 points, which subjectively represents a 70.32% improvement in the quality of life. CONCLUSIONS: Following this therapy the quality of life significantly improved in patients with primarily low quality of life, whereas the group of patients with less affected quality of life reported a temporary decline in the quality of life after the treatment. To obtain a proper objective indication, we suggest a mathematical modelling of possible gains for the individual patient using the University of Washington Quality of Life classification. PMID- 14755421 TI - Photodynamic Therapy of oral dysplasia with topical 5-aminolevulinic acid and light-emitting diode array. AB - BACKGROUND AND OBJECTIVES: In Taiwan, more than two million people have the betel quid (BQ) chewing habit which is a risk factor related to premalignant lesion and squamous cell carcinoma of oral cavity. We developed a light-emitting diode (LED) array combined with topical 5-aminolevulinic acid (ALA) for photodynamic therapy (PDT) and evaluated its effectiveness for the treatment of oral lesions. STUDY DESIGN/MATERIALS AND METHODS: We compared the ALA-PDT effect of the homemade LED array to that of a commercial light source on cultured Ca9-22 human gingival carcinoma cells and the DMBA-induced hamster buccal pouch carcinoma model. Furthermore, we treated several patients having an oral lesion using a topical ALA delivery system and the LED array. RESULTS: The LED array light source was as effective as the commercial light source for ALA-PDT in cultured Ca9-22 cells with LD(50) of 4.5 and 4.3 J/cm(2), respectively, using an MTT assay. This light source was also effective in the DMBA-induced hamster buccal pouch carcinoma model, and in the patients of oral leukoplakia. CONCLUSIONS: ALA-PDT is effective for premalignant lesions such as mucosal dysplasia and carcinoma in situ of oral cavity. Good results could be obtained by using the homemade LED array as light source. The LED array has the advantages of low cost, high reliability, and portability. It is safe, convenient and easy to use for the treatment of oral dysplasia. PMID- 14755422 TI - Investigation of fiber-optic probe designs for optical spectroscopic diagnosis of epithelial pre-cancers. AB - BACKGROUND AND OBJECTIVES: The first objective of this study was to evaluate the performance of fluorescence spectroscopy for diagnosing pre-cancers in stratified squamous epithelial tissues in vivo using two different probe geometries with (1) overlapping versus (2) non-overlapping illumination and collection areas on the tissue surface. Probe (1) and probe (2) are preferentially sensitive to the fluorescence originating from the tissue surface and sub-surface tissue depths, respectively. The second objective was to design a novel, angled illumination fiber-optic probe to maximally exploit the depth-dependent fluorescence properties of epithelial tissues. STUDY DESIGN/MATERIALS AND METHODS: In the first study, spectra were measured from epithelial pre-cancers and normal tissues in the hamster cheek pouch and analyzed with a non-parametric classification algorithm. In the second study, Monte Carlo modeling was used to simulate fluorescence measurements from an epithelial tissue model with the angled illumination probe. RESULTS: An unbiased classification algorithm based on spectra measured with probes (1) and (2), classified pre-cancerous and normal tissues with 78 and 94% accuracy, respectively. The angled illumination probe design provides the capability to detect fluorescence from a wide range of tissue depths in an epithelial tissue model. CONCLUSIONS: The first study demonstrates that fluorescence originating from sub-surface tissue depths (probe (2)) is more diagnostic than fluorescence originating from the tissue surface (probe (1)) in the hamster cheek pouch model. However in general, it is difficult to know a priori the optimal probe geometry for pre-cancer detection in a particular epithelial tissue model. The angled illumination probe provides the capability to measure tissue fluorescence selectively from different depths within epithelial tissues, thus obviating the need to select a single optimal probe design for the fluorescence-based diagnosis of epithelial pre-cancers. PMID- 14755423 TI - Autofluorescence spectrofluorometry of central nervous system (CNS) neuromediators. AB - BACKGROUND AND OBJECTIVES: Changes in the neurotransmitter 5-hydroxytriptamine (5 HT) are related to psychiatric diseases such as depression and anxiety. In this study, 5-HT autofluorescence properties were investigated in solution and in biological tissues. STUDY DESIGN/MATERIALS AND METHODS: Spectrofluorometric characterization was performed on ex vivo samples (tissue sections, homogenates) of the 5-HT-rich brain region hippocampus from rats untreated or treated to affect endogenous 5-HT levels; in vivo, with a 50 solidus in circle optic fiber probe positioned via stereotaxis. RESULTS: 5-HT exhibited minor excitation and emission bands at wavelengths longer than the well known excitation and emission bands in the UV region, 250-320 nm. Spectrofluorometric measurements under 366 nm excitation on homogenates supplied with 5-HT or belonging to treated rats revealed spectral alterations attributable to changes in the amount of 5-HT. Ex vivo and in vivo autofluorescence data were consistent with those obtained by conventional voltammetry. CONCLUSIONS: Autofluorescence spectroscopy potential is confirmed as a suitable technique for the direct measurement of neurotransmitters. PMID- 14755424 TI - MR thermometry-based feedback control of laser interstitial thermal therapy at 980 nm. AB - BACKGROUND AND OBJECTIVES: The goal of this study was to explore the feasibility of magnetic resonance thermal imaging (MRTI)-based feedback control of intracerebral laser interstitial thermal therapy (LITT), using a computer workstation and 980-nm diode laser interfaced to an MR scanner. STUDY DESIGN/MATERIALS AND METHODS: A computer-controlled laser thermal therapy system was used to produce 12 ex vivo lesions in 3 canine and porcine brains and 16 in vivo lesions in 6 canines with diffusing tip fiberoptic applicators and energies from 54 to 900 J. MRTI predictions of thermal damage were correlated with histopathologic analysis. RESULTS: Under feedback control, no carbonization, vaporization, or applicator damage was observed. MRTI-based prediction of thermal dose was not significantly different from histological evaluation of achieved thermal necrosis. CONCLUSIONS: The computer-controlled thermal therapy system was effective at regulating heating, eliminating carbonization and vaporization, and protecting fiberoptic applicators. MRTI estimation of thermal dose accurately predicted achieved thermal necrosis. PMID- 14755425 TI - Infrared imaging of subcutaneous veins. AB - BACKGROUND AND OBJECTIVES: Imaging of subcutaneous veins is important in many applications, such as gaining venous access and vascular surgery. Despite a long history of medical infrared (IR) photography and imaging, this technique is not widely used for this purpose. Here we revisited and explored the capability of near-IR imaging to visualize subcutaneous structures, with a focus on diagnostics of superficial veins. STUDY DESIGN/MATERIALS AND METHODS: An IR device comprising a head-mounted IR LED array (880 nm), a small conventional CCD camera (Toshiba Ik mui, Tokyo, Japan), virtual-reality optics, polarizers, filters, and diffusers was used in vivo to obtain images of different subcutaneous structures. The same device was used to estimate the IR image quality as a function of wavelength produced by a tunable xenon lamp-based monochrometer in the range of 500-1,000 nm and continuous-wave Nd:YAG (1.06 microm) and diode (805 nm) lasers. RESULTS: The various modes of optical illumination were compared in vivo. Contrast of the IR images in the reflectance mode was measured in the near-IR spectral range of 650 1,060 nm. Using the LED array, various IR images were obtained in vivo, including images of vein structure in a pigmented, fatty forearm, varicose leg veins, and vascular lesions of the tongue. CONCLUSION: Imaging in the near-IR range (880-930 nm) provides relatively good contrast of subcutaneous veins, underscoring its value for diagnosis. This technique has the potential for the diagnosis of varicose veins with a diameter of 0.5-2 mm at a depth of 1-3 mm, guidance of venous access, podiatry, phlebotomy, injection sclerotherapy, and control of laser interstitial therapy. PMID- 14755426 TI - Effect of 5-aminolevulinic acid-mediated photodynamic therapy on MCF-7 and MCF 7/ADR cells. AB - BACKGROUND AND OBJECTIVES: Photodynamic therapy (PDT) has been proposed as an alternative approach in overcoming multidrug resistance (MDR) phenotype. To verify whether 5-aminolevulinic acid (ALA)-mediated PDT is effective in MDR cells, we studied the protoporphyrin IX (PpIX) content, intracellular localization, and phototoxicity in human breast cancer cells MCF-7 and derived MDR subline, MCF-7/ADR. STUDY DESIGN/MATERIALS AND METHODS: The fluorescence kinetics of ALA-induced PpIX was evaluated by spectrofluorometer. The phototoxicity of MCF-7 and MCF-7/ADR cells was determined by tetrazolium (MTT) assays and clonogenic assay. Furthermore, Annexin V and propidium iodide (PI) binding assays were performed to analyze the characteristics of cell death after ALA-PDT. RESULTS: MCF-7/ADR accumulated a lower level of PpIX as compared to parental MCF-7 cells. Significant phototoxicity was observed in MCF-7 and increased in a fluence-dependent manner with LD(50) around 8 J/cm(2). Compared to its parental counterpart, MCF-7/ADR cells were less sensitive to ALA photodynamic treatment and PDT-induced cytotoxicity did not increase in a dose responsive manner as the concentration of ALA increased or the fluence of light increased. ALA-PDT was less effective for MCF-7/ADR cells than MCF-7 cells even under the condition when these two cell lines contained the similar amounts of PpIX. CONCLUSIONS: These results indicate that, except for the MDR related characteristics, MCF-7/ADR cells might possess intrinsic mechanisms that render them less sensitive to ALA-PDT induced phototoxicity. PMID- 14755427 TI - Public health approach to birth defects, developmental disabilities, and genetic conditions. PMID- 14755428 TI - Epidemiologic approaches to identifying environmental causes of birth defects. AB - Epidemiology can be used to elucidate environmental causes of birth defects. This paper discusses 1) different types of environmental causes; 2) the difficulties in comparing the prevalence of birth defects between populations, including the need for a population base and the implications of prenatal diagnosis; 3) the main study designs for observational epidemiological studies and the various sources of bias; 4) how statistical power can be increased by meta-analysis or multicentric studies, and improved grouping of birth defects into etiologically more homogeneous subgroups; 5) the distinction between association and causation; 6) the interpretation of clusters in time and space in relation to local environmental causes; and 7) the potential of genetic epidemiology to help elucidate environmental causes. While further research continues into the environmental causes of birth defects, the epidemiologic evidence base for policy making and clinical practice is poor in many areas. The epidemiologic approach is important not only to elucidate environmental causes but also to assess the implementation of existing research into policy and practice for the prevention of birth defects. PMID- 14755429 TI - Vitamin supplements and the risk for congenital anomalies other than neural tube defects. AB - Randomized trials, supported by many observational studies, have shown that periconceptional use of folic acid, alone or in multivitamin supplements, is effective for the primary prevention of neural tube defects (NTDs). Whether this is true also for other congenital anomalies is a complex issue and the focus of this review. It is useful to consider the evidence not only for specific birth defects separately but, importantly, also for all birth defects combined. For the latter, the Hungarian randomized clinical trial indicated, for periconceptional multivitamin use, a reduction in the risk for all birth defects (odds ratio (OR) = 0.53, 95% confidence interval (CI) = 0.35-0.70), even after excluding NTDs (OR = 0.53, 95% CI = 0.38-0.75). The Atlanta population-based case-control study, the only large observational study to date on all major birth defects, also found a significant risk reduction for all birth defects (OR = 0.80, 95% CI = 0.69-0.93) even after excluding NTDs (OR = 0.84, 95% CI = 0.72-0.97). These and other studies also evaluated specific anomalies, including those of the heart, limb, and urinary tract, as well as orofacial clefts, omphalocele, and imperforate anus. For cardiovascular anomalies, two studies were negative, whereas three, including the randomized clinical trial, suggest a possible 25-50% overall risk reduction, more marked for some conotruncal and septal defects. For orofacial clefts, six of seven case-control studies suggest an apparent reduced risk, which could vary by cleft type and perhaps, according to some investigators, by pill dosage. For limb deficiencies, three case-control studies and the randomized trial estimated approximately a 50% reduced risk. For urinary tract defects, three case-control studies and the randomized trial reported reduced risks, as did one study of nonsyndromic omphalocele. All these studies examined multivitamin supplement use. With respect to folic acid alone, a reduced rate of imperforate anus was observed among folic acid users in China. We discuss key gaps in knowledge, possible avenues for future research, and counseling issues for families concerned about occurrence or recurrence of these birth defects. PMID- 14755430 TI - Public health monitoring of developmental disabilities with a focus on the autism spectrum disorders. AB - Developmental disabilities (DDs) are conditions characterized by physical, cognitive, psychological, sensory, adaptive, and/or communication impairments manifested during development. Approximately 17% of individuals in the United States 18 years and younger have a DD, and for most children the cause of their condition is unknown. Of particular interest are the autism spectrum disorders (ASDs), characterized by unusual social, communication, and behavioral development. Previously autism was thought to be a rare condition, but the number of children receiving services for an ASD has increased dramatically in the last decade. Concerns about increases in DDs, particularly ASDs, their causes, and the high costs of intervention have highlighted the need for systematic public health monitoring. Service provider data, such as annual reporting of special education services or of state DD programs, do not provide a complete estimate of the rates for DDs, including ASDs. Unlike genetic metabolic disorders or congenital hearing loss (HL) for which newborn screening programs can provide accurate prevalence rates, there are currently no genetic or biologic markers for the ASDs to enable consistent and early identification of affected children. Centers for Disease Control and Prevention's (CDC) Metropolitan Atlanta Developmental Disabilities Surveillance Program (MADDSP) is a model for population monitoring of ASDs/DDs that has been implemented in other states. This article discusses the role of ASD/DD tracking in public health, as well as the challenges of ASD/DD tracking, including case definition and identification, associated conditions, linkages, and data access. PMID- 14755432 TI - Integration of new genetic diseases into statewide newborn screening: New England experience. AB - Using a data set of newborn screening specimens tested by the New England Newborn Screening Program (NENSP) between January 1999 and February 2003, we analyzed the number of infants with positive newborn screening results and determined how many positive screening results were due to a recent multiplex expansion of services in some of the states. We found that for the subset of the 4-year cohort for which there was a 233% increase in the number of disorders screened (from 9 to 30 disorders), there was a 31% increase in the number of affected infants identified by the screen. We project that if all states in the program expanded their services and if the incidence of disorders is similar across states, there would be an observed 45% increase in the number of infants detected by the screen and a 43% increase in the number of infants for whom the screening algorithm would require some contact with the infants' health care provider. Furthermore, of those requiring contact, we project a 300% increase in the number of screened positive infants who would be referred to tertiary care centers for a diagnostic evaluation. Increased contact with the medical community from additions to newborn screening as demonstrated in this report emphasizes the need for an approach in which the newborn screening program assures coordinated communications between birth units, laboratory, primary health care providers, and specialists. PMID- 14755431 TI - Methodology of a multistate study of congenital hearing loss: preliminary data from Utah newborn screening. AB - A multistate Centers for Disease Control and Prevention (CDC) study was designed to investigate the etiology of congenital hearing loss in infants ascertained through state-mandated hearing screening or early hearing loss detection and intervention (EHDI) programs. At least 50% of permanent childhood-onset hearing loss is due to genetic causes, and approximately 20% of all infants with congenital hearing loss have mutations in the GJB2 gene. Another 1% of childhood hearing loss is due to mitochondrial DNA (mtDNA) mutations. The specific aims of this study are to 1) classify the etiology of congenital hearing loss in infants by doing prospective genetic evaluations of all newborns with permanent hearing loss from defined geographic areas, 2) determine the frequency of mutations in GJB2 and two common mitochondrial mutations in these populations, and 3) establish a model infrastructure linking genetic services to statewide EHDI programs. As of April 2003, Utah is the only center evaluating patients. Study subjects identified through the Utah Department of Health EHDI program are contacted by letter and offered a comprehensive medical genetics evaluation with DNA testing for GJB2 and mitochondrial mutations A1555G and A7445G. To date, 25 probands and their immediate family members have been evaluated. We have identified 20 cases with nonsyndromic hearing loss (7 multiplex and 13 simplex), 4 with syndromic hearing loss, and 1 with presumed cytomegalovirus (CMV)-induced hearing loss. Six of 19 (32%) nonsyndromic cases with sensorineural hearing loss have mutations of one or both alleles of the GJB2 gene, and 21% are homozygous or compound heterozygotes for the 35delG mutation. No A1555G or A7445G mtDNA mutations have been found. Data reported to date include only children born in Utah, but EHDI programs in Hawaii, Rhode Island, and designated areas of Georgia have begun enrolling children in what is now a multistate collaborative study. This is the first comprehensive investigation to determine the etiology of hearing loss from populations ascertained through EHDI programs. The results of this study will facilitate the incorporation of genetic services into EHDI programs. PMID- 14755433 TI - Utility and limitations of genetic disease databases in clinical genetics research: a neurofibromatosis 1 database example. AB - Databases that collect clinical information on patients with particular genetic diseases can be used to investigate the clinical history of a disorder, its genetics, and genotype-phenotype correlations. A database can also serve as a valuable source of patients for studies of disease pathogenesis, variability, or treatment. We review the strengths and limitations of genetic disease databases in the context of our experience with the National Neurofibromatosis Foundation International Database (NNFFID). Genetic disease databases have been developed by individual investigators, scientific consortia, patient support organizations, and commercial enterprises. Databases vary from simple lists of affected individuals to comprehensive collections of detailed clinical and genetic information. Data may be obtained from people who volunteer to be included, systematic assessments of patients seen at participating medical centers, or population-based registries. Access to information may be highly restricted or widely available. These variables all affect the possible uses and usefulness of the data for research. Technical aspects of data entry, organization, storage, and retrieval, as well as issues related to data quality, confidentiality, and security, help determine how well a system actually functions. We discuss examples of research that have been accomplished with genetic disease databases and make recommendations regarding the organization and operation of these resources. PMID- 14755434 TI - Contribution of Mendelian disorders to common chronic disease: opportunities for recognition, intervention, and prevention. AB - Recognizing Mendelian disorders should improve health care for persons with strong familial risks for common chronic diseases. The Online Mendelian Inheritance in Man (OMIM) database was reviewed to identify Mendelian disorders featuring 17 common chronic diseases, including 9 cardiovascular conditions, diabetes, and 7 common cancers. Mendelian disorders were selected if any one of the 17 diseases was reported in more than two families manifesting in adulthood. Patterns of chronic diseases and modes of inheritance associated with these Mendelian disorders are described. The GeneTests/Reviews database and other websites were reviewed to determine availability of genetic testing and management and prevention recommendations for the selected disorders. Of 2,592 (OMIM) entries reviewed, 188 Mendelian disorders were selected. Most (67.7%) are autosomal dominant disorders. Almost half (45.8%) feature combinations of the chronic diseases under study. At least one gene is known for 68.8% of the selected disorders, and clinical genetic testing is available for 55% of disorders. Guidelines for management and prevention are available for 33.9% of these, ranging from recommendations for supportive care to guidelines for managing affected persons and screening relatives. Significant clinical heterogeneity exists for Mendelian disorders that might present as strong family histories of common chronic diseases. Recognition of the different combinations of diseases within a pedigree, including mode of inheritance and heritable disease risk factors, facilitates diagnosis of these Mendelian disorders. Genetic testing is available for most disorders, which can further clarify the genetic risk, and for some, recommendations for management and prevention are available. However, evidence-based guidelines are needed. PMID- 14755435 TI - Ethical issues concerning genetic testing and screening in public health. AB - Genetic testing (predictive analysis that determines genetic alterations in individuals for clinical purposes) and screening (programs that identify persons within a subpopulation who may be at a higher risk for a genetic disease or condition) are increasingly utilized to promote and improve the public's health. The proliferate use of genetic testing and screening may improve public health outcomes, but it also implicates significant ethical, legal, and social concerns. Within the context of conflicting ethical values from the individual and public health perspectives, individual values such as informed consent and privacy and discrimination protections must be respected. Legal and ethical attempts to exceptionalize genetic tests and information (as compared to other health information) to protect privacy and prevent discrimination are well intended, but can also be unjust and impractical. Respect for individual ethical rights has limits. Principles of public health ethics justify voluntary genetic testing and screening and sharing of data for population-based health purposes. Thus, individual rights should not always trump the use of genetic tests or screening programs (or information derived therefrom) for legitimate public health purposes. PMID- 14755436 TI - Nongenetic pathologic developments of brain-wave patterns in monozygotic twins discordant and concordant for schizophrenia. AB - Evidence from previous studies has suggested that the inter-individual differences in human brain-wave patterns (EEG) are predominantly determined by genetic factors. In particular, the within-pair EEG concordance of monozygotic (mz) twins was found to be typically as high as r = 0.81 across channels and frequency bands, thus being comparable to that between repeated assessments on the same individual with typically r = 0.83. Yet our investigations into mz twins discordant and concordant for schizophrenia yielded a significantly reduced within-pair EEG concordance for both, the pairs discordant for schizophrenia and the pairs concordant for schizophrenia (with concordance for schizophrenia assessed through a syndrome-oriented approach). A multivariate discriminant function of EEG parameters distinguished in a reproducible way between affected and unaffected subjects at an overall performance of >75% correctly classified subjects, while the severity of illness, as derived from EEG-differences between affected and unaffected subjects, was closely related to the severity of illness as provided by psychopathology syndrome scores. Consequently, EEG anomalies associated with schizophrenia and manifested differently in the mz co-twins concordant for schizophrenia are likely the effect of nongenetic, pathologic processes that evolved independently in the co-twins' genetically identical brains once the illness began to progress. The existence of such nongenetic processes would suggest a modification of the standard phenotype-to-genotype search strategies of molecular-genetic studies that aim to link the schizophrenia phenotype to genetic vulnerability factors. PMID- 14755437 TI - Multiple missense mutations in the diazepam binding inhibitor (DBI) gene identified in schizophrenia but lack of disease association. AB - The diazepam binding inhibitor (DBI), alternatively known as the acyl-CoA binding protein (ACBP), is involved in multiple biological actions. The polypeptide binds to the peripheral, or mitochondrial, benzodiazepine receptor and facilitates transport of cholesterol to the inner membrane to stimulate steroid synthesis. Through this action, DBI indirectly modulates gamma-aminobutyric acid (GABA) mediated inhibitory neurotransmission. DBI can be postulated as a candidate gene for psychiatric phenotypes including anxiety, mood, and psychotic disorders. In an examination of the DBI gene among 112 individuals with schizophrenia, our laboratory has identified 18 novel single nucleotide polymorphisms (SNPs), including three missense changes in conserved amino acids, a coding region microdeletion, and multiple SNPs in the putative promoter region. Case-control association analyses were performed for the missense changes, but none was found to be significantly associated with disease. PMID- 14755438 TI - No evidence of association or linkage disequilibrium between polymorphisms in the 5' upstream and coding regions of the dopamine D4 receptor gene and schizophrenia in a Portuguese population. AB - Alterations in dopaminergic system have been implicated in the pathophysiology of this disease for many years, and this study was performed to assess the possible involvement of the dopamine D4 receptor (DRD4) gene polymorphisms either in the 5' upstream or in the coding regions, in the etiology of schizophrenia. The approach included an association study with 90 Portuguese trios by doing the analysis of the individual alleles and the haplotypes. For the polymorphisms in the 5' upstream region (-C616G and -C521T) and in the coding region (48 bp repeat) of the DRD4 gene, negative results were obtained with both haplotype relative risk (HRR) and transmission disequilibrium test (TDT), as well as transmit. These data suggest that polymorphisms (-C616G, -C521T, and 48 bp repeat) at the DRD4 gene do not have a minor effect in the susceptibility to schizophrenia in our sample. PMID- 14755439 TI - CALCYON gene variation, schizophrenia, and cocaine dependence. AB - Calcyon is a brain-specific D1 dopamine receptor-interacting protein, with a potential role in D1-mediated physiological processes, including motor control, reward mechanisms, and cognitive processes. Our objective was to investigate the relationship between polymorphism of the CALCYON gene and (1) schizophrenia and (2) cocaine dependence in African-American (AA) and European-American (EA) subjects. Two single nucleotide polymorphisms (SNPs) at the CALCYON locus were genotyped in 70 AA and 206 EA individuals with schizophrenia and 90 AA and 118 EA individuals with cocaine dependence. The control group was comprised of 46 AA and 207 EA subjects screened to exclude those with psychiatric or substance use disorders. The specific polymorphisms studied were markers +295214G/A and +297151T/G. Comparisons of allele and haplotype frequencies between cases and controls were performed with the Fisher's Exact Test. Linkage disequilibrium (LD) between these two SNPs was calculated with the 3LOCUS program. No alleles or haplotypes were found to be associated with schizophrenia or cocaine dependence either in AA or EA subjects. The markers +295214G/A and +297151T/G are in the same haplotype block in all subgroups. Allele and haplotype frequencies differed significantly between EA and AA subjects. These results suggest that these two genetic variants in the CALCYON gene do not play a major role in predisposition to either schizophrenia or cocaine dependence in AA or EA subjects. Furthermore, these findings begin to establish a haplotype map for this gene in the AA and EA populations. PMID- 14755440 TI - Is the WKL1 gene associated with schizophrenia? AB - A missense mutation Leu309Met in the WKL1 (MLC1, KIAA0027) gene, mapped to 22q13.3, was reported to co-segregate with periodic catatonic schizophrenia (SCZ) in a single large German pedigree with seven affected individuals (Meyer et al. [2001: Mol Psychiatry 6:302-306]). This report raised the following questions that were dealt with in the present study: does the Leu309Met mutation have a role in SCZ, or only in catatonic SCZ? Does the mutation Leu309Met in the WKL1 gene, encoding a putative membrane protein, non-selective cation channel, have any effect on the channel activity? Is the WKL1 gene, which is expressed exclusively in brain, expressed differently in SCZ brains compared to controls? These questions were answered by screening the Leu309Met mutation in 117 Israeli Jewish patients with SCZ (55 Ashkenazi and 62 non-Ashkenazi Jews) and 176 matched controls. In search of differences in the level of WKL1 gene expression, postmortem dorsalateral prefrontal cortex of 16 schizophrenic patients and 15 controls was checked. We also measured the putative channel activity of normal WKL1 subcloned in pcDNA3 to determine the effect of the reported Leu309Met mutation. Our results argue against the involvement of WKL1 in SCZ susceptibility. PMID- 14755441 TI - Polymorphisms in the dopamine D5 receptor (DRD5) gene and ADHD. AB - There is considerable evidence to support a role of dopamine-related genes in the molecular aetiology of attention-deficit hyperactivity disorder (ADHD). A microsatellite located near the dopamine D5 receptor (DRD5) gene has been associated with ADHD in a number of studies, but other polymorphisms within the vicinity of this gene have not been examined. In this study we genotyped three microsatellites spanning the DRD5 region in a large clinical sample. Overall, we found little evidence to support a role for DRD5 in ADHD. We found no evidence of association with either the previously associated DRD5 marker, or a repeat in the promoter region of the gene. We did, however, find significant association for an allele of D4S615, a dinucleotide repeat located 131 kb 3' of DRD5 that has been previously associated with schizophrenia. A global test incorporating all alleles of this marker, however, was not significant and thus this finding needs replication before any conclusions can be made. PMID- 14755442 TI - TNXB locus may be a candidate gene predisposing to schizophrenia. AB - We report here on the detection of nine single nucleotide polymorphisms (SNPs) near to the NOTCH4 locus in the search for schizophrenia susceptibility genes in the class III region of the human major histocompatibility complex (MHC). We totally analyzed 122 family trios recruited in the UK. The TDT analysis demonstrated that of the nine SNPs, three were associated with schizophrenia, including rs1009382 (P = 0.00047), rs204887 (P = 0.007), and rs8283 (P = 0.015). Both rs1009382 and rs204887 are present in the TNXB locus. The rs1009382 is a non synonymous SNP located in exon 23 of the gene and its A to G base change causes a Glu2578Gly substitution. The goodness-of-fit test showed that genotypic distribution of rs1009382 was deviated from Hardy-Weinberg equilibrium due to homozygote excess in the patient group (P = 0.01), suggesting that a double dose of a genetic risk may be involved. Possibly, rs1009382 is a candidate SNP predisposing to a schizophrenic illness. Moreover, the test for linkage disequilibrium (LD) between paired SNPs showed that the nine SNPs studied may be in the same LD block with an unexpected pattern as the strength of LD was not correlated with the distance between paired SNPs. The haplotype analysis suggested that there might be more than one disease-related allele located in the class III region of the MHC, and that these alleles possibly confer either susceptibility or resistance to schizophrenia. PMID- 14755443 TI - Confirming RGS4 as a susceptibility gene for schizophrenia. AB - A recent study identified a putative association between variants in the regulator of G-protein signalling 4 (RGS4) and schizophrenia, Chowdari et al. [2002: Hum Mol Genet 11: 1373-1380]. RGS4 is both a positional and functional candidate gene for schizophrenia. Chowdari and colleagues identified association at this locus in a number of distinct and ethnically diverse samples, although the pattern of association was not the same in all the samples. Our study attempted to replicate this association in an independent Irish sample of schizophrenia cases and controls. We succeeded in detecting evidence of association at the RGS4 locus. The signal comes from a four-marker haplotype that is in significant excess in our case sample. The same haplotype is in excess in the Caucasian schizophrenia sample used by Chowdari et al. [2002: Hum Mol Genet 11: 1373-1380]. This study provides further support for the contribution of RGS4 to schizophrenia susceptibility. PMID- 14755444 TI - Association and transmission analysis of the FMR1 IVS10 + 14C-T variant in autism. AB - Evidence from the high male to female ratio of individuals with autism as well as suggestive linkage data have implicated the possible involvement of X chromosomal loci in the aetiology of autism. Studies of the FMR1 gene on Xq27 have shown that occasionally individuals, and particularly females, with the [CGG] repeat expansion and methylation mutation may present with autistic symptoms. However, molecular studies suggest that such mutations are not a major cause of autism. Previously, we have screened autism probands for mutations in regions of the FMR1 gene downstream of the [CGG] repeat and identified an intronic variant in the FMR1 gene, IVS10 + 14C-T, which was present at a significantly higher frequency in autistic individuals compared to controls individuals. We have now investigated this variant in a further 136 autism families and 186 control individuals. We have established that the variant is significantly more frequent among East Asian individuals within our affected and control groups (P < 0.0001) and although we observed a trend of higher transmission frequency of the rare allele to affected individuals, there was no significant evidence in either family-based or case/control association studies for this variant in autism (P > 0.05). PMID- 14755445 TI - No association between the APOE gene and autism. AB - Autism is a neurodevelopmental disorder characterized by stereotypic and repetitive behavior and interests, together with social and communicative deficiencies. The results of several genomic screens suggest the presence of an autism susceptibility locus on chromosome 19p13.2-q13.4. The apolipoprotein E (APOE) gene on chromosome 19 encodes for a protein, apoE, whose different isoforms (E2, E3, E4) influence neuronal growth. APOE participates in lipid transport and metabolism, repair, growth, and maintenance of axons and myelin during neuronal development. The APOE protein competes with the Reelin protein for VLDL/APOER2 receptor binding. Several studies have reported evidence for an association between autism and the Reelin gene. Based on these data we tested for association between APOE and autism using family-based association methods in a data set of 322 autism families. Three promoter, one intronic, and one 3' UTR single nucleotide polymorphisms (SNPs) in the APOE gene (-491a/t, -427c/t, 219g/t, 113c/g, and 5361c/t) as well as the APOE functional polymorphism (E2, E3, E4) were examined and failed to reveal significant evidence that autism is associated with APOE. PMID- 14755446 TI - Notch4 gene polymorphisms are not associated with autism in Japanese population. PMID- 14755447 TI - Association of tryptophan 2,3 dioxygenase gene polymorphism with autism. AB - Although elevation of blood and platelet serotonin has been documented in autism, genetic analyses of serotonin transporter gene have given conflicting results. Tryptophan 2,3 dioxygenase (TDO2) is the rate-limiting enzyme in the catabolism of tryptophan, the precursor of serotonin. A mutation that results in decreased activity of the TDO2 can decrease catabolism of tryptophan and increase the level of whole body serotonin. As such it is a potential candidate gene for autism. We have investigated five single nucleotide polymorphisms in the TDO2 gene for association with autistic disorder. One hundred and ninety six multiplex autistic disorder families were tested using transmission disequilibrium test. There was a significant difference in the transmission of a promoter variant to autistic subjects (P = 0.0006). Haplotype analysis also demonstrated significant difference in the transmission of TDO2 haplotypes to autistic subjects (P = 0.0027). Our results suggest the presence of a susceptibility mutation in the TDO2 or a nearby gene, but may also represent a chance finding. PMID- 14755448 TI - Clinical features of psychotic disorders and polymorphisms in HT2A, DRD2, DRD4, SLC6A3 (DAT1), and BDNF: a family based association study. AB - Schizophrenia is clinically heterogeneous and multidimensional, but it is not known whether this is due to etiological heterogeneity. Previous studies have not consistently reported association between any specific polymorphisms and clinical features of schizophrenia, and have primarily used case-control designs. We tested for the presence of association between clinical features and polymorphisms in the genes for the serotonin 2A receptor (HT2A), dopamine receptor types 2 and 4, dopamine transporter (SLC6A3), and brain-derived neurotrophic factor (BDNF). Two hundred seventy pedigrees were ascertained on the basis of having two or more members with schizophrenia or poor outcome schizoaffective disorder. Diagnoses were made using a structured interview based on the SCID. All patients were rated on the major symptoms of schizophrenia scale (MSSS), integrating clinical and course features throughout the course of illness. Factor analysis revealed positive, negative, and affective symptom factors. The program QTDT was used to implement a family-based test of association for quantitative traits, controlling for age and sex. We found suggestive evidence of association between the His452Tyr polymorphism in HT2A and affective symptoms (P = 0.02), the 172-bp allele of BDNF and negative symptoms (P = 0.04), and the 480-bp allele in SLC6A3 (= DAT1) and negative symptoms (P = 0.04). As total of 19 alleles were tested, we cannot rule out false positives. However, given prior evidence of involvement of the proteins encoded by these genes in psychopathology, our results suggest that more attention should be focused on the impact of these alleles on clinical features of schizophrenia. PMID- 14755449 TI - Evidence of a founder effect in families with frontotemporal dementia that harbor the tau +16 splice mutation. AB - The +16 exon 10 splice mutation of the tau gene (microtubule-associated protein tau, MAPT) has been reported in numerous families with frontotemporal dementia (FTD). To date, the majority of these families are from England and Wales in the UK, although families with this mutation have been reported from Australia and the USA. Our own analysis has identified eight families with the +16 MAPT splice mutation from around the Manchester and North Wales areas of the UK. Given the proximity of the UK families to one another it is likely that they are related and represent a single extended pedigree. In order to investigate this possibility, and the possibility that the families with this mutation from London, the USA, and Australia are related, we genotyped 11 microsatellite markers around the tau locus. In most cases (20/25, 80%), a common haplotype, approximately 3 cM in size, was identified. In the remaining cases, this haplotype appears to have been varyingly reduced in size by recombination. We demonstrate that the +16 mutation is on the H1 tau haplotype and that H1 specific polymorphisms are also shared by these families. These data provide evidence that the MAPT +16 splice mutation cases from around the world analyzed in this study are indeed related and represent a single pedigree that probably originated in the North Wales area of the UK. Furthermore, this single large pedigree may be of use in the identification of disease modifying loci in FTD. PMID- 14755450 TI - Genetic mapping using haplotype and model-free linkage analysis supports previous evidence for a locus predisposing to severe bipolar disorder at 5q31-33. AB - We report further evidence for our previous suggestion [Garner et al., 2001: Am J Hum Genet 68:1061-1064] of a locus on 5q predisposing to bipolar I disorder (BP I) in an extended Costa Rican pedigree. We genotyped additional microsatellite markers in this region and applied a multi-point non-parametric linkage analysis (SimWalk2). Significant identity-by-descent allele sharing among affected relatives was observed for all of the 20 markers tested in a segment of approximately 15 cM. Most affected individuals shared a single haplotype over this region; breaks within this haplotype may suggest a more restricted candidate location for a BP-I gene. These results support the suggestion that a locus at 5q31-33, together with a previously reported locus at 18q22-23, may provide the major genetic risk for BP-I in this family. PMID- 14755451 TI - No association between the insulin degrading enzyme gene and Alzheimer's disease in a Japanese population. AB - Susceptibility to Alzheimer's disease (AD) is thought to be regulated by multiple genetic factors. Recently, three independent studies have reported that loci on chromosome 10q are linked with AD, and the insulin degrading enzyme (IDE; MIM 146680) gene located on chromosome 10q23-q25; IDE is located close to the maker D10S583, which exhibits a maximum LOD score for late-onset AD. We examined seven polymorphisms in the IDE gene, the marker D10S583 in the 5' flanking region, and SNPs in introns 1, 3, 11, 20, 21, and 22 (rs#1999764, 1855915, 1970244, 538469, 551266, and 489517, respectively). Four SNPs in introns 3, 11, 20, and 22 did not exhibit any polymorphisms in the Japanese population that was studied. D10S583 and two SNPs in introns 1 and 21 did not exhibit a significant association with early- or late-onset AD. In addition, no associations were observed for subgroups of AD grouped according to APOE status. The present study indicates that the IDE gene polymorphisms do not confer susceptibility to early- or late-onset AD at least in a Japanese population. PMID- 14755452 TI - The gender effect in juvenile Huntington disease patients of Italian origin. AB - We analyzed a population of juvenile Huntington disease (HD) subjects of Italian origin (n = 57). The main aim of this study was to analyze the gender effect of the affected parent on age at onset and clinical presentation of offspring with juvenile HD. We also analyzed molecular features of the disease, including CAG mutation length and GluR6 gene polymorphism, according to the affected parent's gender. The mutation length was longer in paternally than in maternally transmitted HD juvenile patients (P = 0.025), nevertheless a similar mean early onset in the two groups (P > 0.05). This data was even enforced by that obtained from the whole cohort of patients included in the databank (n = 600) where, in the presence of increased mean parent-child CAG repeat change in paternal vs. maternal meiotic transmissions (+7.3 vs. +0.7 CAG, P = 0.0002), the mean parent child year-of-onset change was similar in the two groups (-10.4 and -7.0 years, P > 0.05). A lower TAA-triplet in GluR6 was associated with an earlier age at onset in juvenile patients (P = 0.031, R2 = 0.10). When we added the GluR6 effect on age at onset to the CAG expanded number effect (P = 0.0001, R2 = 0.68) by multiple regression approach, the coefficient of determination R2 increased to 0.81. This effect in addition to the expanded CAG repeat number, found in juvenile and not in adult patients, was slightly enforced by paternal compared to maternal transmissions (R2=0.82). Our findings suggest the occurrence of a weaker effect of the paternal mutation on juvenile age at onset in our population, possibly amplified by other genetic factors, such as the TAA-triplet length in the GluR6 gene. PMID- 14755453 TI - An investigation into sub-telomeric deletions of chromosome 22 and pervasive developmental disorders. AB - Deletions of the sub-telomeric region of chromosome 22 have been associated with mental retardation, developmental delay, and autistic behaviors. This study investigated sub-telomeric anomalies of chromosome 22 using fluorescent in situ hybridization (FISH) probes in 82 subjects diagnosed with autism and atypical autism. No microdeletions were detected in this group. Similar FISH analyses were undertaken on two children with developmental delay, who were ascertained to be ring 22 during routine cytogenetic investigations. One subject was shown to have a microdeletion in the sub-telomeric region tested. Both children met the social and communication cut off for autism on the ADI and but did not meet the cut off for restrictive and repetitive behaviors. Only one of the two children met the criteria for PDD on the ADOS. PMID- 14755454 TI - Association of adverse childhood experiences, age of menarche, and adult reproductive behavior: does the androgen receptor gene play a role? AB - Previous research has reported associations between adverse childhood experiences, early menarche, and early sexual activity. One hypothesis to account for these findings is that an X-linked androgen receptor GGC-repeat polymorphism predisposes fathers to behaviors which include family abandonment and their daughters to earlier menarche and sexual activity and less stable relationships. Retrospective data relevant to this theory were examined from a community survey involving 3,702 women in the age groups 20-24, 40-44, and 60-64 years, and another involving 908 women aged 18-79 years. Earlier age of menarche was found to be associated with adverse childhood experiences and earlier sexual activity. However, the androgen receptor gene polymorphism was unrelated to adverse fathering behavior or to marital breakdown. PMID- 14755455 TI - A dyslexia susceptibility locus (DYX7) linked to dopamine D4 receptor (DRD4) region on chromosome 11p15.5. AB - Dyslexia is a disability in acquiring reading and spelling skills that is independent of general intelligence and educational opportunity, and is highly heritable. It is known that dyslexia often co-occurs with attention deficit hyperactivity disorder (ADHD), and the 7-repeat allele of the 48-bp tandem repeat in exon 3 of the dopamine D4 receptor (DRD4) has been implicated in ADHD. We, therefore, investigated DRD4 as a candidate gene for dyslexia by testing for linkage and association with 14 markers at and around the DRD4 locus on chromosome 11p15.5. Using 100 families having at least two siblings affected with dyslexia, model-free linkage analysis revealed evidence for linkage to the DRD4 exon 3 repeat (two-point MFLOD = 2.27, P = 0.001) and to HRAS located just proximal to DRD4 (two-point MFLOD = 2.68, P = 0.0004). Evidence for linkage was maximal between DRD4 and HRAS (three-point MFLOD = 3.57, P = 0.00005). However, linkage disequilibrium analysis showed no significant evidence for association between dyslexia and DRD4 or HRAS. In particular, dyslexic subjects showed no significant increase of the DRD4 7-repeat allele associated with ADHD. It is possible that other DRD4 variants, not in strong linkage disequilibrium with the exon 3 repeat polymorphism, or alternatively, another gene very closely linked to DRD4, may influence susceptibility to dyslexia. PMID- 14755456 TI - Haplotypes of the monoamine oxidase genes and the risk for substance use disorders. AB - Monoamine oxidase A (MAOA) locus is an attractive candidate for exploring genetic contribution to the variation in the risk for substance use disorders (SUD) because of its important role in the metabolism of neurotransmitters, including dopamine and serotonin. Prior findings have suggested an association of the MAOA gene with the risk for early onset SUD. To extend this research, we genotyped four MAOA markers (two VNTR polymorphisms and two SNPs) and built a cladogram reflecting the evolutionary history of MAOA haplotypes [Nguyen et al., under review]. The cladogram served as the framework for nested ANOVA and logit analyses of association between MAOA and indices of liability to SUD (diagnosis, age of onset, and a dimensional index of substance use related problems) in a sample of adult males of European ancestry. Whereas no association was found for the categorical diagnosis, a significant relationship was detected between the dimensional liability indices and MAOA haplotypes. Overall, our results, albeit not definitive, are consistent with the hypothesis that variants in MAOA account for a small portion of the variance of SUD risk, possibly mediated by liability to early onset behavioral problems. PMID- 14755457 TI - Association study of cannabinoid receptor gene (CNR1) alleles and anorexia nervosa: differences between restricting and binging/purging subtypes. AB - Anorexia nervosa (AN) is a severe and disabling psychiatric disorder, characterized by profound weight loss and body image disturbance. Family and twin studies indicate a significant genetic contribution to this disorder although no genetic mutation has yet been identified. The endocannabinoid system has recently been implicated in many physiological functions including appetite regulation. We, therefore, undertook a family based study to test the hypothesis whether a polymorphism of the CNR1 gene, which encodes human CB1 receptor, a subclass of the central cannabinoid receptor, contributes to the susceptibility to AN. Fifty two families (parents with one or two affected siblings) were genotyped for the (AAT) trinucleotide repeat of CNR1 gene. Using the haplotype relative risk (HRR) method, the distribution of alleles transmitted to the patients was not found to be significantly different from the non-transmitted parental alleles. However, upon dividing the samples to restricting and binging/purging subtypes of AN, the extended transmission disequilibrium test (ETDT) revealed that there is preferential transmission of different alleles in each of the subtypes. The 14 repeat allele was preferentially transmitted in the binging/purging AN group (P = 0.05) but not in the restricting AN group, whereas the 13 repeat allele was preferentially transmitted in the restricting AN group (almost significant, P = 0.07) but not in the binging/purging AN group. Our study suggests that restricting AN and binging/purging AN may be associated with different alleles of the CNR1 gene. PMID- 14755458 TI - Mutations in the optineurin gene in Japanese patients with primary open-angle glaucoma and normal tension glaucoma. AB - The optineurin gene (OPTN) was identified as a gene that causes primary open angle glaucoma (POAG) and normal tension glaucoma (NTG). To investigate the frequency of sequence changes in OPTN in Japanese glaucoma patients, single strand conformation polymorphism analysis and subsequent sequence analysis were performed for genotyping OPTN in 165 unrelated Japanese patients with POAG and 148 patients with NTG, with 196 control subjects without glaucoma as reference subjects. Out of four mutations reported to be associated with risk and to cause disease in Caucasian patients, sequence alterations in 458G > A and 691_692insAG were not detected in any investigated Japanese patients with glaucoma, and alterations in 1944G > A and 603T > A, were present in similar frequencies in glaucoma patients and control subjects. The current results suggest that there may be certain racial differences between Japanese and Caucasians with respect to OPTN genotypes. PMID- 14755459 TI - Screening adherence in BRCA1/2 families is associated with primary physicians' behavior. AB - This study provides an assessment of long-term breast and ovarian cancer screening behaviors and the variables associated with adherence with screening among women with or at 50% for having a BRCA1/2 mutation. Participants in the study included 112 women (33 mutation carriers and 79 at 50% risk). Data was collected through a mailed questionnaire, which included items to assess screening behaviors in the last 2 years, risk perception, cancer specific distress, adherence determinants, specific barriers, and cancer history. Statistical analysis included descriptive statistics and non-parametric tests to describe bivariate associations and regression analysis. Adherence rates were 72% for annual mammography, 21% for semi-annual clinical breast exam (CBE), 29% for monthly breast self-exam (BSE), and 19% for annual transvaginal ultrasound (US). Only one participant was adherent with semi-annual CA125. Variables that had a significant association (P < 0.05) with at least one screening modality included: a lack of time, marital status, education, cancer history, provider concern, perceived screening utility, confidence in ability to overcome barriers, cancer specific distress, and risk perception. Primary physician behavior, either in terms of screening recommendations or screening performance in the case of CBE, had significant independent association with adherence to mammography, CBE, and US screening recommendations. The results of this study highlight the essential role that primary physicians play in supporting their very high-risk patients' adherence. PMID- 14755460 TI - Septo-optic dysplasia and amniotic bands: further evidence for a vascular pathogenesis. AB - Septo-optic dysplasia (SOD) is a heterogeneous disorder. While most cases represent an isolated defect, SOD has also been seen in association with mutations in single genes, as a part of multiple congenital anomaly syndromes and with exposure to various teratogens. We report a boy with features of SOD who also has limb defects suggestive of amniotic bands. This case, in addition to others in the literature, provides evidence for a vascular pathogenesis of SOD in some individuals. This hypothesis is also supported by the sporadic occurrence of SOD and its association with decreased maternal age, vascular teratogens, and neuropathologic findings suggestive of vascular insults. PMID- 14755461 TI - Targeted scan of fifteen regions for nonsyndromic cleft lip and palate in Filipino families. AB - Cleft lip with or without cleft palate (CL/P) is a congenital anomaly with variable birth prevalence based on geographic origins, with the highest rates commonly found in Asian populations. About 70% of cases are nonsyndromic (NS), in which the affected individual has no other abnormalities. NS CL/P is a complex disorder with genetic and environmental effects and no specific genetic loci yet confirmed. Fifteen candidate regions were examined for linkage to NS CL/P. Regions were chosen based on previous suggestive linkage and/or association in human families, or suggestive animal model data. Polymorphic markers in these regions were genotyped for analysis on 36 Filipino families comprised of 126 affected and 218 unaffected individuals. An additional 70 families with 149 affecteds were used for replication of suggestive results. Parametric (LOD score) and nonparametric (SIMIBD) linkage analyses were performed as well as transmission disequilibrium test (TDT) analysis. Five markers yielded suggestive results from the 36 families. The parametric LOD scores for the MSX1-CA and D4S1629 were >1.0 and the SIMIBD P values for D6S1029 and RFC1 are suggestive (<0.06), while the SIMIBD P value of 0.01 for TGFA was significant. Since the Msx1 mouse knockout has cleft palate and MSX1 mutations have been found in rare cases of syndromic CL/P, this locus is especially plausible for linkage. Previous studies have also found linkage of NS CL/P to 4q31 and 6p23. These regions contain several candidate genes, including AP2 at 6p23 and FGF2, BMPR1B, and MADH1 at 4q31. TGFA has both linkage and linkage disequilibrium data supporting it as a candidate gene for NS CL/P. While no region was definitively confirmed for linkage to NS CL/P, the data do support further investigation using larger sample sizes and candidate gene studies at 2p13.2, 4p16.2, 4q31, 6p23, and 16q22 24. PMID- 14755462 TI - Association of chromosomal regions 3p21.2, 10p13, and 16p13.3 with nonsyndromic cleft lip and palate. AB - Approximately 4,000 babies with nonsyndromic cleft lip with or without cleft palate (NSCLP) are born each year in the United States. Because NSCLP exhibits both etiologic and genetic heterogeneity, attempts to identify the underlying genetic causes have met with limited success and the pursuit of early promising findings have yielded mixed results. Two recent genomic scans identified a number of suggestive regions; some of these results have been supported by our lab and others in subsequent studies. Using our NSCLP multiplex family population, we were able to provide additional supportive evidence for association to the regions 2q37, 11p12-14, 12q13, and 16p13.11-p12 that were identified in the genomic scans. However, there remains a number of additional viable candidate genes and regions that have not been sufficiently investigated. These include chromosomal translocations in patients with NSCLP, growth factor genes, metalloproteinase (MMP) and transcription factor (patterning) genes, including those in the WNT family. Here, we present results from screening the 10p13 chromosomal translocation region associated with NSCLP, MMP genes clustered on chromosomes 1p36, 11q22.3, 16p13.3, and 16q12-13, and the region containing the WNT5A gene on chromosome 3p21. Markers from three of the regions, 10p13, 16p13.3 (MMP25), and 3p21.2, yielded findings that are sufficiently significant to warrant closer investigation. PMID- 14755463 TI - Nonsyndromic cleft lip and palate: four chromosomal regions of interest. AB - Nonsyndromic cleft lip with or without cleft palate (NSCLP), a common birth defect affecting 1/700 live births and 4,000 newborns/year in the United States, is associated with short and long-term morbidity. As such, it has significant impact on the health care system. NSCLP is a complex disorder that results from the interaction of genetic and environmental factors that are slowly being defined. Genomic scans have suggested a number of regions that may contain NSCLP loci. In this study, we have evaluated regions identified by a previously published genomic scan of affected sib-pairs and have found evidence of association in sixty-five multiplex families for eight chromosomal regions. Four of these regions, 2q37, 11p12-14, 12q13, and 16p13, have also been identified in second genomic scan of multiplex families from China. One region, defined by D11S1392, gave evidence suggesting the presence of an NSCLP locus in all three studies. Altogether, these results suggest chromosomal regions that should be targeted in order to identify NSCLP loci. PMID- 14755464 TI - Physical activity and angiotensin-converting enzyme gene polymorphism in mild hypertensives. AB - It has been suggested that the insertion(I) allele of the I/deletion(D) polymorphism of the angiotensin converting enzyme (ACE) gene is associated with endurance exercise and increased physical conditioning in response to this type of exercise. To investigate the association between the ACE I/D polymorphism and physical activity status in 355 never treated, stage I hypertensives (265 men, 90 women, mean age: 33 +/- 9 years), in whom power exercise is contraindicated, participants of the HARVEST study. Physical activity was assessed using a standardized questionnaire. BMI and age did not vary among genotypes. None of active subjects performed power oriented exercises. ACE I/D frequencies (II-18%, ID-55%, DD-27%) were in Hardy-Weinberg equilibrium. Sedentary lifestyle was more common among DD than II hypertensives (76% in DD, and 48% in II, Chi(2) = 13.9, P = 0.001). In stepwise MANOVA using age, marital status, profession, sex, and ACE genotype as predictors of physical activity, marital status (F = 24.4, P < 0.0001) and ACE genotype (F = 16.03, P < 0.0001) contributed to more than 50% of the variance in physical activity status of the population. Our results suggest that the ACE I/D polymorphism may be a specific genetic factor associated with physical activity levels in free-living borderline and mild hypertensive subjects. PMID- 14755465 TI - X-linked spondyloepiphyseal dysplasia tarda: a novel SEDL mutation in a Jewish Ashkenazi family and clinical intervention considerations. AB - X-linked spondyloepiphyseal dysplasia tarda (SEDT; MIM 313400) is a late onset progressive skeletal disorder, which manifests in childhood and is characterized by disproportionate short stature with a short trunk, barrel chest and absence of systemic complications. We found a single-nucleotide deletion in position 613 of the SEDL gene in two brothers of Jewish-Ashkenazi ancestry afflicted with the disease. This is the first description of SEDL mutations in a Jewish family. Following this finding, an eight-month old second cousin of the brothers, who had yet no clinical or radiological signs of the disease, was found to carry the deletion. Another relative, 24-years old, carrying the same mutation was 1.61 m tall and had only minimal signs of the disease. These findings raise the dilemma of pre-natal counseling in SEDL and the need for exploring means of early intervention in pre-symptomatic cases. PMID- 14755466 TI - A new type of autosomal recessive spondyloepiphyseal dysplasia tarda. AB - Repeated occurrence of a hitherto unrecognized form of spondyloepiphyseal dysplasia tarda (SED tarda) has been studied in two independent families. Because parental consanguinity was also present in one family, autosomal recessive inheritance is proposed. The onset was in late childhood. The slowly evolving disorder shared several features of the already known types of SED tarda. The radiographic abnormalities were limited to the spine and proximal femora. The patients' hands were normal. The entity described is set apart not only from the X-linked and autosomal-dominant forms of SED tarda but also from the already delineated autosomal recessive types by significant clinical and radiographic differences. Final genotypic characterization must await the results of genetic linkage studies and of appropriate molecular genetics investigations. PMID- 14755467 TI - Post-natal short stature, short limbs, brachydactyly, facial abnormalities, and delayed bone age: a new syndrome? AB - A boy is described with clinical features including post-natal short stature, short limbs, speech delay, relatively large skull, prominent metopic ridge, wide palpebral fissures, proptosis, epicanthic folds, capillary hemangioma between the eyes and nose, flat nasal bridge, everted and small nares, small ears with a narrow external auditory canal and thick lobes, short neck, brachydactyly, bilateral Simian creases, a single flexion crease of the first and fifth fingers, protruding abdomen, and bilateral cryptorchidism. Radiographs did not show any features of bone dysplasia. However, a delayed bone age was noted. The boy's parents are first cousins. To the best of our knowledge, this constellation of anomalies has not been reported before, and may be considered a new syndrome. PMID- 14755468 TI - Spondyloepimetaphyseal dysplasia of Maroteaux (pseudo-Morquio type II syndrome): report of a new patient and review of the literature. AB - An 11-year-old girl was seen with short stature, a head positioned in hyperextension, mild arched palate, prominent joints, limited elbow movements, hyperextensible wrists and fingers, brachydactyly, broad thorax, pectus carinatum, short trunk, a genu valgum, and flat feet. A radiographic skeletal survey revealed a generalized osteoporosis, platyspondyly, thoracic kyphoscoliosis, small and square iliac wings, short femoral necks, dysplastic epiphyses, flared metaphyses and brachydactyly with various carpal, metacarpal, and finger malformations. These features are very close to a very rare entity: the spondyloepimetaphyseal dysplasia (SEMD) of Maroteaux or "pseudo-Morquio" type II syndrome, whose specific radiological characteristics are found in this case. PMID- 14755469 TI - Submicroscopic deletion 9(q34.3) and duplication 19(p13.3): identified by subtelomere specific FISH probes. AB - Submicroscopic rearrangements involving chromosome ends are responsible for the unexplained mental retardation and multiple congenital anomalies observed in a number of patients. We have studied a patient with mental retardation, significant microcephaly, alopecia universalis, and other anomalies who carries an unbalanced segregant from a cryptic reciprocal translocation involving chromosomes 9 and 19. FISH studies using subtelomere specific probes revealed a derivative chromosome 9 in which the 9q subtelomeric sequence has been replaced by 19p subtelomeric sequence. As a result, the patient has partial monosomy 9q and partial trisomy 19p. The patient inherited the derivative 9 from his father, who carries a cryptic apparently balanced reciprocal translocation involving the terminal regions of 9q and 19p. This case is exceptional in that reports of rearrangements involving distal chromosome 9q and 19p are rare. This study demonstrates the utility of subtelomere specific FISH probes for detecting cryptic subtelomeric rearrangements in patients with idiopathic mental retardation and normal appearing karyotypes. PMID- 14755470 TI - Boy with 47,XXY,del(15)(q11.2q13) karyotype and Prader-Willi syndrome: a new case and review of the literature. AB - We report on a 10-year-old boy with a 47,XXY,del(15)(q11.2q13) karyotype and a Prader-Willi syndrome phenotype. His medical history and physical examination conformed to all of the major clinical criteria for Prader-Willi syndrome, but his height was taller than expected based on his hand and foot sizes. The deleted chromosome 15 was paternal in origin and molecular analysis showed maternal origin for the additional X chromosome. These findings suggest that the presence of these two disorders was coincidental in our patient. This supports the findings in the two other 47,XXY and Prader-Willi cases for which parent of origin studies have been published. Given the information from the literature and presented herein, we suggest that genetic counseling for cases of PWS and 47,XXY should address these two conditions separately. PMID- 14755471 TI - Two cousins with partial trisomy 12q and monosomy 12p recombinants of a familial pericentric inversion of the chromosome 12. AB - Partial trisomy 12q and monosomy 12p lead to multiple malformation syndromes. Instead of trisomy 12q that has been reported as a clinically identifiable syndrome, monosomy 12p is characterized by a wide phenotypic spectrum. We report two cousins suffering from severe mental retardation, seizures, and dysmorphic features related to a trisomy 12q24.3-->qter and a monosomy 12p13-->pter resulting from a familial pericentric inversion of chromosome 12. In an attempt to improve the clinical delineation of these two syndromes, we compared our two patients with previous reports of these aneusomies. This review emphasizes the high frequency of familial translocations, including a breakpoint at 12q24 involved in trisomy 12q whereas monosomy 12p occurs most frequently de novo. Despite the poor specificity of the signs, this comparison allowed us to determine the clinical features present in more than 20% of patients with trisomy 12q or monosomy 12p. We particularly emphasize some consistent leading features of monosomy 12p, including microcephaly, dental, cardio-vascular, extremity, and sensorial abnormalities, initially not reported as recurrent in this syndrome. PMID- 14755472 TI - Unbalanced cryptic 5p deletion/17p duplication identified by subtelomeric FISH in a family with a boy with chimerism and a balanced t(4;5). AB - The use of subtelomeric FISH probes has greatly supplemented conventional chromosome analysis in detecting cryptic anomalies in patients with mental retardation (MR), dysmorphic features, and congenital malformations. We report a 3-month-old boy who was diagnosed with ambiguous genitalia, dysmorphic features, and developmental delay. Standard chromosome studies on blood revealed a chimeric karyotype of 46,XY,t(4;5)(q31.1;q14)[46]/46,XX[4]. The boy had intra-abdominal gonads that were testicular in origin by biopsy. Multiple dysmorphic features, marked hypotonia, developmental delay, poor growth, and relative macrocephaly were noted on physical exam. His 2.5-year-old sister also presented with hypotonia, developmental delay, relative macrocephaly, and similar dysmorphic stigmata. In addition, she was diagnosed with several internal malformations. Her karyotype was 46,XX. Due to the striking phenotypic similarity, subtelomeric FISH studies were initiated in the siblings. In addition to the known balanced karyotypic abnormalities, the boy was found to have a derivative chromosome 5 with a 5pter deletion and a 17pter duplication. This cryptic abnormality was also detected in his sister. Chromosome analysis of the father revealed a subtle balanced t(5;17)(p15.31;p13.1) which was confirmed by subtelomeric FISH, whereas the mother's chromosome complement was normal. This familial constellation illustrates the usefulness of subtelomeric FISH in the diagnosis of cryptic chromosome abnormalities in patients for whom conventional karyotype does not disclose findings sufficient to explain the observed phenotypic anomalies. PMID- 14755473 TI - A case of Yunis-Varon syndrome complicated with complete cleft lip and palate. PMID- 14755474 TI - A 23-year-old woman with Down syndrome, type 1 neurofibromatosis, and breast carcinoma. PMID- 14755475 TI - Diaphragmatic hernia in 18p- syndrome. PMID- 14755476 TI - Distal arthrogryposis in two sisters born to different fathers. PMID- 14755477 TI - Atypical phenotype and intrafamilial variability associated with a novel SALL1 mutation. PMID- 14755478 TI - Turner syndrome phenotype with 47,XXX karyotype: further investigation warranted? PMID- 14755479 TI - 47,XXX associated with malformations. PMID- 14755481 TI - Electrodiagnostic studies in the management and prognosis of neuromuscular disorders. AB - Prognosis remains a neglected aspect of modern medical care and research, behind diagnosis and treatment. The very term "electrodiagnosis" implies as much. Despite this, much has been published regarding the use and benefit of electrodiagnostic techniques in assessing prognosis and assisting in management of patients after the diagnosis has been established. This information is often hidden or otherwise not emphasized. This review summarizes the literature regarding the use of such techniques for prognosis and management of disorders of lower motor neurons, peripheral nerves, neuromuscular transmission, and muscle. PMID- 14755482 TI - Matrix metalloproteinases and skeletal muscle: a brief review. AB - Matrix metalloproteinases (MMPs) are a family of zinc- dependent proteolytic enzymes that function mainly in the extracellular matrix, where they contribute to the development, functioning, and pathology of a wide range of tissues. This mini-review describes the MMPs and tissue inhibitors of MMPs (TIMPs) in skeletal muscle, and considers their involvement in muscle development, ischemia, myonecrosis, angiogenesis, denervation, exercise-induced injuries, disuse atrophy, muscle repair and regeneration, and inflammatory myopathies and dystrophies. Despite the very limited information currently available on MMPs and their inhibitors in skeletal muscle, it is becoming increasingly clear that they have important physiological functions in maintenance of the integrity and homeostasis of muscle fibers and of the extracellular matrix. Understanding the roles of MMPs and TIMPs may lead to the development of new drug-related treatments for various muscle disorders based on suppression or upregulation of their expression. PMID- 14755483 TI - Electrodiagnostic services in the United States. AB - To identify patterns of service provision in the United States, electrodiagnostic encounters were characterized by analyzing data from the 1998 MarketScan Commercial Claims & Encounters Database (The MEDSTAT Group). Electrodiagnostic related claims were identified using CPT codes for nerve conduction studies, electromyography, and somatosensory evoked potentials. Over 130,000 electrodiagnostic claims representing 48,207 separate electrodiagnostic encounters were identified. Neurologists accounted for the highest percentage of physician providers (23.6%), followed closely by physiatrists (22%). Physiatrists performed the services at a higher rate than any other provider (886.3 studies per 100 practicing physiatrists). Electrodiagnostic encounters by nonphysician providers accounted for 16.9% of all studies. Physical therapists were the dominant nonphysician providers of electrodiagnostic services (9.3%), followed by podiatrists (5.5%). Physician providers had a median of three claims per encounter compared with one claim per encounter for nonphysician providers. The relatively large proportion of studies conducted by nonphysicians, as well as their limited extent of testing, raises concerns about the quality of care for these patients. These quality issues should be addressed in future studies. PMID- 14755484 TI - Loss-of-function phenotype of hereditary neuropathy with liability to pressure palsies. AB - Hereditary neuropathy with liability to pressure palsies (HNPP) provides a human model to investigate the role of PMP22 in myelinated peripheral nerve, since the disease is caused by a deletion of one of the two PMP22 alleles. To systematically characterize the phenotype of HNPP, we prospectively evaluated the clinical features and electrophysiological findings in 17 genetically confirmed patients, 7 men and 10 women, ranging in age from 9 to 66 years (mean, 41 +/- 13). Fifteen symptomatic patients presented with episodes of transient focal weakness or sensory loss that were usually related to particular activities causing nerve compression, including stretching or minor repetitive focal trauma. No patient sought medical attention for symptoms of a symmetric polyneuropathy. Neurological examinations were either normal or mildly abnormal. Neither focal slowing of nerve conduction studies, nor reduction in compound muscle action potential (CMAP) or sensory nerve action potential (SNAP) amplitudes consistently predicted the site of symptoms. We conclude that the majority of patients with HNPP present with transient, recurrent, focal symptoms of weakness or sensory loss in the distribution of individual nerves or plexus, and that a diffuse symmetric sensorimotor polyneuropathy is an unusual presentation of HNPP. These studies suggest that the function of PMP22, at least in part, is to stabilize myelin so that it will be protected from injuries resulting from repetitive, minor trauma. PMID- 14755485 TI - Reproducibility of statistical motor unit number estimates in amyotrophic lateral sclerosis: comparisons between size- and number-weighted modifications. AB - Motor unit number estimations (MUNEs) can directly assess motor unit populations in muscle and quantify the degree of physiological or pathological motor unit degeneration. A high degree of reproducibility and reliability is required of any effective quantitative tool. MUNE is being increasingly applied clinically, and statistical MUNE has several advantages over alternative techniques. Nevertheless, the optimal method of applying statistical MUNE with respect to its reproducibility has not been established. We performed statistical MUNE by selecting the most compensated compound muscle action potential (CMAP) area as a test area and modified the results obtained by using the weighted mean surface recorded motor unit potential (SMUP). MUNE measurements made in patients with amyotrophic lateral sclerosis (ALS) showed better reproducibility after incorporating the size-weighted modification. Therefore, we suggest that the size weighted MUNE in combination with the selection of testing "neurogenically compensated" CMAP areas is a more reliable method of statistical MUNE analysis in ALS patients. PMID- 14755486 TI - Transplantation of primary satellite cells improves properties of reinnervated skeletal muscles. AB - Skeletal muscle demonstrates a force deficit after repair of injured peripheral nerves. We tested the hypothesis that transplantation of satellite cells into reinnervated rabbit tibialis anterior (TA) muscles improves their properties. Adult rabbits underwent transection and immediate suture of the common peroneal nerve. In order to provide an environment favorable for cell transplantation, TA were then made to degenerate by cardiotoxin injection, either immediately or after a 2-month delay, which is sufficient for muscle reinnervation. In both cases, the injured TA were transplanted with cultured satellite cells 5 days after induction of muscle degeneration. When cells were transferred immediately after nerve repair, drastic morphological and functional muscle alterations were observed. However, when the muscles were allowed to become reinnervated before cell transplantation, muscles were heavier and developed a significantly higher maximal force compared to denervated-reinnervated muscles. Thus, application of the cell therapy protocol improved properties of denervated muscles only when they were allowed to become innervated. These results, which represent the application of cell therapy to improve force recovery of reinnervated muscles, will be of significant interest in certain clinical contexts, particularly after immediate or delayed muscle reinnervation. PMID- 14755487 TI - Excitability properties of human median axons measured at the motor point. AB - Threshold tracking was used to measure excitability indices (strength-duration properties, threshold electrotonus, and the current-threshold relationship) at the motor point of the abductor pollicis brevis, and the results were compared with those of the median nerve at the wrist. Using an accelerometer placed at the thumb tip, movement-related potentials were recorded as target responses. When stimulating at the same site, excitability measurements were no different, and their variability no greater, when the target responses were movements rather than muscle action potentials. Motor point stimulation resulted in significantly shorter strength-duration time-constant and higher rheobase than wrist stimulation. In addition, the technique of latent addition showed that a slow component was much smaller at the motor point than at the wrist. In threshold electrotonus, threshold changes in response to depolarizing and hyperpolarizing conditioning currents were significantly smaller at the motor point than at the wrist. The differences in strength-duration time-constant and latent addition suggest that persistent Na(+) current at the resting potential is smaller at the motor point. The differences in threshold electrotonus may depend in part on altered fiber geometry but suggest that inward and possibly outward rectification are increased distally. Motor point excitability testing may provide new insights into the pathophysiology of the nerve terminals in a variety of peripheral neuropathies and motor neuron disorders. PMID- 14755488 TI - Passive exercise and fetal spinal cord transplant both help to restore motoneuronal properties after spinal cord transection in rats. AB - Spinal cord transection influences the properties of motoneurons and muscles below the lesion, but the effects of interventions that conserve muscle mass of the paralyzed limbs on these motoneuronal changes are unknown. We examined the electrophysiological properties of rat lumbar motoneurons following spinal cord transection, and the effects of two interventions shown previously to significantly attenuate the associated hindlimb muscle atrophy. Adult rats receiving a complete thoracic spinal cord transection (T-10) were divided into three groups receiving: (1) no further treatment; (2) passive cycling exercise for 5 days/week; or (3) acute transplantation of fetal spinal cord tissue. Intracellular recording of motoneurons was carried out 4-5 weeks following transection. Transection led to a significant change in the rhythmic firing patterns of motoneurons in response to injected currents, as well as a decrease in the resting membrane potential and spike trigger level. Transplants of fetal tissue and cycling exercise each attenuated these changes, the latter having a stronger effect on maintenance of motoneuron properties, coinciding with the reported maintenance of structural and biochemical features of hindlimb muscles. The mechanisms by which these distinct treatments affect motoneuron properties remain to be uncovered, but these changes in motoneuron excitability are consistent with influences on ion conductances at or near the initial segment. The results may support a therapeutic role for passive limb manipulation and transplant of stem cells in slowing the deleterious responses of motoneurons to spinal cord injury, such that they remain more viable for subsequent alternative strategies. PMID- 14755489 TI - Temporal discrimination threshold on various parts of the body. AB - The temporal discrimination threshold (TDT) of various parts of the body was investigated in 35 healthy volunteers, and the effect of aging on the TDT was studied in 80 subjects (aged 18-82 years). Ascending (ATDT) and descending (DTDT) TDT values were measured in 13 areas using a pair of electrical stimuli. Both ATDT and DTDT differed significantly among the body parts (P < 0.01, one-way repeated ANOVA), and the TDT was shortest on the index finger and longest on the lower leg, where it was approximately 156% of that on the finger. There was no difference of the TDT value with gender or between sides. There was no effect of aging on the TDT in subjects aged 18-64 years, but the value was prolonged in subjects over 65 years. We suggest that the TDT difference among body parts is mainly due to the difference in sensory processes in the central nervous system, and that it may provide information about changes in the system related to aging. PMID- 14755490 TI - Effect of static stretch training on neural and mechanical properties of the human plantar-flexor muscles. AB - To determine the contributions of neural and mechanical mechanisms to the limits in the range of motion (ROM) about a joint, we studied the effects of 30 sessions of static stretch training on the characteristics of the plantar-flexor muscles in 12 subjects. Changes in the maximal ankle dorsiflexion and the torque produced during passive stretching at various ankle angles, as well as maximal voluntary contraction (MVC) and electrically induced contractions, were recorded after 10, 20, and 30 sessions, and 1 month after the end of the training program. Reflex activities were tested by recording the Hoffmann reflex (H reflex) and tendon reflex (T reflex) in the soleus muscle. Training caused a 30.8% (P < 0.01) increase in the maximal ankle dorsiflexion. This improved flexibility was associated (r(2) = 0.88; P < 0.001) with a decrease in muscle passive stiffness and, after the first 10 sessions only, with a small increase in passive torque at maximal dorsiflexion. Furthermore, both the H- and T-reflex amplitudes were reduced after training, especially the latter (-36% vs. -14%; P < 0.05). The MVC torque and the maximal rate of torque development were not affected by training. Although the changes in flexibility and passive stiffness were partially maintained 1 month after the end of the training program, reflex activities had already returned to control levels. It is concluded that the increased flexibility results mainly from reduced passive stiffness of the muscle-tendon unit and tonic reflex activity. The underlying neural and mechanical adaptation mechanisms, however, showed different time courses. PMID- 14755491 TI - Prediction of aspiration in myasthenia gravis. AB - Prediction of the risk of dysphagia and aspiration is important in the management of myasthenia gravis (MG). We assessed the ability of four bedside clinical tools to predict aspiration in 20 MG patients. Patients completed a self-directed questionnaire, underwent clinical neurological assessment and a bedside speech pathology assessment, and were assessed with the quantitative myasthenia gravis (QMG) score. The ability of these tools to predict aspiration was compared with the results of a modified barium swallow. Seven patients aspirated, 4 silently. The total self-directed questionnaire score, two specific questions on the self directed questionnaire, the prediction based on clinical neurological assessment, and the QMG bulbar subset score all correlated with aspiration. The speech pathology prediction was highly sensitive but less specific. This pilot study shows that simple clinical tools can predict which MG patients are at risk of aspiration. PMID- 14755492 TI - Overexpression of semicarbazide-sensitive amine oxidase in human myopathies. AB - Oxidative stress has been implicated in the pathogenesis of several muscle diseases. Semicarbazide-sensitive amine oxidase (SSAO) metabolizes oxidative deamination of primary aromatic and aliphatic amines. In the oxidative reactions, amine substrates are converted into the aldehyde, followed by the production of ammonia and H(2)O(2). Although normal levels in muscle are very low, SSAO is expressed in almost all mammalian tissues. In this study, we examined the possible implication of SSAO as an additional source of oxidative stress in the pathogenesis of muscle disorders. The expression of SSAO was examined immunohistochemically in muscle biopsy specimens from patients with inclusion body myositis (IBM; n = 5), desmin-related myopathy (DRM; n = 3), dermatomyositis (n = 3), granulomatous (sarcoid) myopathy (n = 2), muscle denervation reinnervation (n = 3), and rhabdomyolysis (n = 2), as well as from control subjects (n = 3). Strong SSAO immunoreactivity was present in vacuolated and nonvacuolated fibers in IBM, in abnormal fibers in DRM, and in degenerating and regenerating fibers in dermatomyositis and rhabdomyolysis. In addition, SSAO overexpression was observed in muscle fibers adjacent to granulomas in sarcoid myopathy. These results suggest that SSAO is a source of oxidative stress in diseased human skeletal muscle and that it contributes to oxidative stress induced damage in various inflammatory and other myopathies. Alternatively, the expression of SSAO in muscle fibers may be a consequence of muscle fiber injury. PMID- 14755494 TI - Muscle pathology in 57 patients with myotonic dystrophy type 2. AB - We evaluated muscle biopsies from 57 patients with genetically confirmed myotonic dystrophy type 2/proximal myotonic myopathy (DM2/PROMM). Light microscopy showed myopathic together with "denervation-like" changes in almost all biopsies obtained from four different muscles: increased fiber size variation, internal nuclei, small angulated fibers, pyknotic nuclear clumps, and predominant type 2 fiber atrophy. Quantitative morphometry in 18 biopsies that were immunostained for myosin heavy chain confirmed a predominance of nonselective type 2 fiber atrophy. These histological changes were similar in all patients regardless of the site of biopsy, the predominant clinical symptoms and signs, and the clinical course. It is likely that, in a number of undiagnosed patients, DM2 is the underlying disorder. With a better understanding of the histopathological pattern in DM2, biopsies from patients with undiagnosed neuromuscular disorders can now be reevaluated. PMID- 14755493 TI - Quadriceps muscle endurance in patients with chronic obstructive pulmonary disease. AB - The purpose of this study was to compare quadriceps muscle strength (maximal voluntary contraction, MVC), endurance (T(LIM)), and dynamic work capacity (W(LIM)) in 89 patients (57 men and 32 women) with chronic obstructive pulmonary disease (COPD) to 31 healthy controls (20 men and 11 women). Compared with the healthy subjects, COPD patients demonstrated statistically significant (P < 0.05) lower values for MVC, T(LIM) and W(LIM). The MVC was 20-30% lower, whereas T(LIM) and W(LIM) were 70-80% lower in the patients. Stepwise multiple regression analysis revealed that pulmonary function variables contributed only partially to the explanation of variance in MVC, T(LIM), and W(LIM). Our results indicate that in patients with COPD, quadriceps muscle endurance and dynamic work capacity are impaired to a greater extent than is quadriceps muscle strength. For rehabilitation purposes, the findings may lead to more specific exercise programs, further enhancing rehabilitation outcomes in terms of functional abilities and social participation. PMID- 14755495 TI - Reproducibility of muscle-fiber conduction velocity estimates using multichannel surface EMG techniques. AB - The aim of this study was to assess the reproducibility of muscle-fiber conduction velocity (MFCV) estimates obtained from multichannel surface electromyographic (EMG) recordings. Surface EMG signals were collected with a matrix of 61 electrodes during isometric, submaximal (50% of the maximal voluntary contraction torque) contractions of the biceps brachii muscle. Conduction velocity was estimated using multichannel maximum likelihood techniques. Reproducibility of MFCV estimates was assessed varying the number of signals (two to seven) used for the estimate and the distance between detection points (5-30 mm). Intraclass correlation coefficient (ICC) of both initial MFCV values and their rates of change with fatigue increased when increasing number of signals and distance between detection points. ICC of initial MFCV was negative using two signals for MFCV estimate, and it increased to approximately 75% with six to seven signals. Thus, reproducibility of MFCV estimates may be improved significantly using advanced multichannel estimation methods with respect to classic two-channel techniques. PMID- 14755496 TI - EMG and nerve conduction studies in children with congenital muscular dystrophy. AB - Motor and sensory nerve conduction velocities (NCVs) and needle electromyography (EMG) results were reviewed in 26 children with different types of congenital muscular dystrophy (CMD), including patients with mutations in the genes LAMA2, FKRP, and COL6A2. In every patient, at least one EMG examination detected myopathic changes that were predominant in proximal muscles, although EMG performed at birth was normal in two patients. Brief bursts of high-frequency repetitive discharges were electrically elicited in four patients. Uniformly slowed motor NCVs without signs of denervation were observed in seven patients: five merosin-deficient, one merosin-positive, and one with unavailable merosin status. The merosin-deficient neuropathy also involved sensory nerves in three patients and worsened with age in two. In conclusion, myopathic EMG changes were typical and early findings in all types of CMD. An associated neuropathy was detected in most patients with merosin-deficient CMD, and also in a child with normal merosin expression. PMID- 14755497 TI - Static load magnitude is a risk factor in the development of cumulative low back disorder. AB - Occupations requiring frequent periods of static lumbar flexion are known epidemiologically to be risk factors for the development of cumulative low back disorder. The impact of the load magnitude sustained during a series of short static lumbar flexions followed by an equally long rest period on the development of a cumulative low back disorder was addressed in an in vivo feline model. Static loads of 20, 40, and 60 N were applied over 10 min of flexion followed by 10-min rest sessions that were repeated six times (for a total of 2 h) while monitoring lumbar viscoelastic creep (laxity) and reflex electromyographic (EMG) activity from the multifidus muscles. Creep and EMG were also monitored over 7 h of rest following the six flexion-rest sessions. It was found that the creep developed in the 10-min flexion periods did not recover completely during the following 10 min of rest, giving rise to a large cumulative creep at the end of the work-rest session. Muscle activity demonstrated spasms during the static flexion periods as well as initial and delayed hyperexcitability during the 7-h rest period. Loads of 20 and 40 N did not result in delayed hyperexcitability, whereas loads of 60 N resulted in delayed hyperexcitability. Statistical analysis demonstrated that increased load significantly intensified the magnitude of the hyperexcitabilities (P < 0.05). Thus, repeated periods of static lumbar flexion were found to result in a transient neuromuscular disorder with an intensity directly related to the load magnitude, which should be considered a compounding risk factor. PMID- 14755498 TI - Somatosensory evoked potentials for the electrodiagnosis of meralgia paresthetica. AB - The aim of this study was to evaluate the diagnostic utility of somatosensory evoked potential (SSEP) studies of the lateral femoral cutaneous nerve (LFCN) in assessing patients with meralgia paresthetica. Twenty-one consecutive patients with unilateral meralgia paresthetica, as defined clinically (sensory impairment of lateral aspect of the thigh) and electrodiagnostically (abnormal sensory nerve conduction), and 21 control subjects were studied with two SSEP methods. SSEPs were elicited by stimulation of the LFCN below the anterior superior iliac spine (ASIS stimulation) and by cutaneous stimulation of the lateral aspect of the distal third of the thigh (thigh stimulation). Abnormalities were defined by the SSEP interside latency difference, interside amplitude ratio, or an absent response. The SSEP with ASIS stimulation had a sensitivity of 5% and a specificity of 95%, whereas with thigh stimulation it had a sensitivity of 52% and a specificity of 76%. Overall, SSEP after ASIS stimulation had no diagnostic value. Recording of the SSEP after thigh stimulation is recommended in obese patients only when sensory nerve conduction cannot be determined. PMID- 14755499 TI - Regulatory light chain phosphorylation increases eccentric contraction-induced injury in skinned fast-twitch fibers. AB - During contraction, activation of Ca(2+)/calmodulin-dependent myosin light chain kinase (MLCK) results in phosphorylation of myosin's regulatory light chain (RLC), which potentiates force and increases speed of force development over a wide range of [Ca(2+)]. We tested the hypothesis that RLC phosphorylation by MLCK mediates the extent of eccentric contraction-induced injury as measured by force deficit in skinned fast-twitch skeletal muscle fibers. Results indicated that RLC phosphorylation in single skinned rat psoas fibers significantly increased Ca(2+) sensitivity of isometric force; isometric force from 50 +/- 16 to 59 +/- 18 kN/m(2) during maximal Ca(2+) activation; peak absolute power output from 38 +/- 15 to 48 +/- 14 nW during maximal Ca(2+) activation; and the magnitude of contraction-induced force deficit during maximal (pCa 4.5) activation from 26 +/- 9.8 to 35 +/- 9.6%. We conclude that RLC phosphorylation increases force deficits following eccentric contractions, perhaps by increasing the number of force generating cross-bridges. PMID- 14755500 TI - Transthyretin amyloidosis presenting with multifocal demyelinating mononeuropathies. AB - We describe a patient with transthyretin amyloidosis who presented with multifocal mononeuropathies with features of demyelination on nerve conduction studies, a constellation of findings not previously described in amyloid polyneuropathy. Genetic testing revealed a valine122isoleucine mutation in the coding region of the transthyretin gene, a mutation that generally presents with late-onset cardiac amyloidosis. Our patient was also unusual in that she was 34 years old at the time of presentation and had no cardiac involvement. PMID- 14755501 TI - Adult polyglucosan body disease: case description of an expanding genetic and clinical syndrome. AB - A non-Jewish patient is described who had adult polyglucosan body disease (APBD) and glycogen branching enzyme (GBE) deficiency without GBE mutation. A heterozygous polymorphism (Val160Ile) was found, and also discovered in 1 of 50 normal individuals. Magnetic resonance imaging demonstrated increased T2 signal in the midbrain, medullary olives, dentate nuclei, cerebellar peduncles, and internal and external capsules, with vermian atrophy. Both muscle and nerve biopsy revealed perivascular inflammatory infiltrates. These findings expand the clinical and genetic spectrum of APBD. Factors other than mutation of the expressed GBE gene may cause enzyme deficiency and varied expression and development of APBD. PMID- 14755502 TI - Axial myopathy in myasthenia: a misleading cause of dropped head. PMID- 14755503 TI - Monophasic negative sympathetic skin responses and autonomic dysfunction in carpal tunnel syndrome. PMID- 14755505 TI - Toxicity of cytoreductive surgery and hyperthermic intra-peritoneal chemotherapy. AB - BACKGROUND AND OBJECTIVES: Cytoreduction with hyperthermic intra-peritoneal chemotherapy (HIPEC) is a treatment with a high morbidity. Optimal patients selection can possible reduce toxicity and complications. PATIENTS AND METHODS: Complications and toxicity of 102 patients were studied. Toxicity was graded according National Cancer Institute Common Toxicity Criteria (NCI CTC) classification. A complication was defined as any post-operative event that needed re-intervention. Potential patients, tumor, and treatment factors were studied on their relation to complications. RESULTS: Grade 3, 4, or 5 toxicity was observed in 66 patients (65%). Eight patients died of treatment-related causes. Surgical complications occurred in 36 patients (35%). Fistulae were frequently encountered (18 patients). The risk of a complicated recovery was higher in carcinomatosis with recurrent colorectal cancer (P = 0.009) and in the case of more than five regions affected (P = 0.044), who had a Simplified Peritoneal Cancer (SPC) score of 13 (P = 0.012) and with an incomplete initial cytoreduction (P = 0.035). Patients with blood loss exceeding 6 L (P = 0.028) and those with three or more anastomoses also had an increased post-operative complication rate (P = 0.018). CONCLUSIONS: Toxicity of cytoreduction followed by HIPEC was 65% (Grade 3-5 NCI CTC), with a surgical complication rate of 35%. Patients with six or seven regions involved and those in whom complete cytoreduction cannot be reached are probably better off without this treatment. PMID- 14755506 TI - The prognostic significance of margin width for extremity and trunk sarcoma. AB - BACKGROUND AND OBJECTIVES: To evaluate the significance of resection margin for soft tissue sarcoma (STS), we determined local recurrence-free interval (LRFI), distant metastases-free interval (DMFI), and overall survival (OS) for primary extremity and truncal STS with clear margins (> or =10 mm), close margins (1-9 mm), and positive margins (0 mm). METHODS: Patients were evaluated via review of charts and tumor specimens. RESULTS: Among 111 patients, tumors were predominantly high grade (86%), > or =5 cm (76%), and deep (81%). A minority of patients received adjuvant radiation (38%) and/or adjuvant chemotherapy (34%). Margin width was > or =10 mm (48%), 1-9 mm (40%), or 0 mm (12%). Margins > or =10 mm were less common for large (P = 0.009) or deep (P = 0.02) tumors. By multivariate analysis, independent factors for LRFI were tumor size (P = 0.04) and margin width (P = 0.03). Independent factors related to DMFI were tumor grade (P = 0.002), size (P = 0.007), and patient age (P = 0.02). Independent factors relating to OS were tumor grade (P = 0.001), size (P = 0.004), and depth (P = 0.03). CONCLUSIONS: Margins > or =10 mm independently predicted longer LRFI and are optimal for extremity STS resection. Adjuvant radiotherapy should be considered for all STS resected with margins <10 mm, and margin width should be considered when reporting and interpreting LR outcomes for these patients. PMID- 14755508 TI - Low sensitivity of the touch imprint cytology of the sentinel lymph node in breast cancer patients--results of a large series. AB - BACKGROUND AND OBJECTIVES: Touch imprint cytology (TIC) was reported to be a sensitive method of intraoperative sentinel lymph node (SLN) assessment. The objective of our study was to assess the value of the TIC as an intraoperative SLN evaluation method and to determine a subgroup of patients in whom TIC should not be indicated. METHODS: In 250 breast cancer patients with SLN biopsy, TIC of SLNs was performed intraoperatively. The results of TIC were compared to the final histopathological analysis of SLNs. A subgroup analysis of the TIC value was performed with regard to the tumor size. RESULTS: SLN metastases were found in 102/250 patients (41%). Two cases were false positive. The sensitivity of TIC was 34%, specificity 98.6%, accuracy 72%, negative predictive value 69%, and positive predictive value 95%. TIC was significantly more sensitive to detect macrometastases (32/43) compared to micrometastases or ITC (3/59) (P < 0.001). TIC was positive in only 5% (4/77) in T < 10 mm tumors compared to 18% (31/168) in T > 10 mm (P = 0.01). CONCLUSIONS: TIC is a simple, quick, and sensitive method of intraoperative SLNs evaluation for the presence of the macrometastases. TIC has a very limited value in detecting micrometastases and no value in detecting ITC. TIC may not be indicated in T1a + b tumors. PMID- 14755507 TI - Feasibility of sentinel node biopsy for breast cancer after neoadjuvant endocrine therapy: a pilot study. AB - BACKGROUND AND OBJECTIVES: Sentinel node biopsy (SNB) is used for evaluation of axillary lymph node status of patients with breast cancer. The usefulness of SNB after neoadjuvant chemotherapy is not established. In addition, SNB after endocrine therapy has rarely been evaluated. We assessed the feasibility of dye guided SNB after neoadjuvant endocrine therapy in comparison with those of SNB after chemotherapy. METHODS: A total of 36 patients subjected to SNB after endocrine therapy alone (n = 16) (tamoxifen, anastrozole or combination of goserelin and tamoxifen) or after chemotherapy (n = 20) (anthracycline and/or taxane) were included. SNB was performed with indigocarmine dye prior to the wide resection of the tumor or mastectomy. RESULTS: Sentinel nodes were successfully identified in 100% (16/16) of patients after endocrine therapy and in 85% (17/20) of patients after chemotherapy. The mean number of harvested sentinel nodes was 2.2 after endocrine therapy and 1.8 after chemotherapy. There was no false negative case after endocrine therapy and there was one false negative case after chemotherapy (8% of overall false negative rate). CONCLUSIONS: SNB seemed feasible and accurate after neoadjuvant endocrine therapy in patients with breast cancer. PMID- 14755510 TI - End-to-end anastomosis using a surgical instrument after a segmental resection of the distal duodenum. PMID- 14755511 TI - Intraoperative cholangioscopy in the management of biliary adenomas. PMID- 14755512 TI - Inferiorly based rectus abdominis myocutaneous flaps in surgical oncology: Indications, technique, and experience in 37 patients. PMID- 14755513 TI - Bromodeoxyuridine administered during neurogenesis of the projection neurons causes cerebellar defects in rat. AB - Bromodeoxyuridine (BrdU) is broadly used in neuroscience to study embryonic development and adult neurogenesis. The potential toxicity of this halogenated pyrimidine analogue is frequently neglected. In this study, we administered BrdU in small doses by the progressively delayed cumulative labeling method to immunocytochemically tag different cerebellar cell types with antibodies to specific markers and BrdU in the same section. The well-known structure of the cerebellum made it possible to ascertain several toxic effects of the treatment. Time-pregnant rats were given five or six injections of 5 or 6 mg of BrdU ( approximately 12-20 mg/kg) at 8-hour intervals over 2 successive days between day 11 and 21 of pregnancy (E11-E12 to E20-E21), and the adult progeny was processed by immunocytochemistry. We demonstrate that this treatment effectively labeled distinct cerebellar cell populations but produced striking defects in the proliferation, migration, and settling of the Purkinje cells; reduced the size of the cerebellar cortex and nuclei; produced defects in the patterning of foliation; and also affected litter size, body weight, and mortality of the offspring. The observed toxic effects were consistent within individual treatment groups but varied between different treatment groups. Treatment with BrdU at the peak of neurogenesis of cerebellar projection neurons (E14) produced the most severe malformations. We observed no overt effects on the timing of neurogenesis for cerebellar neurons and glia across experimental groups. In conclusion, BrdU is a useful tool to study neural development, but its cytotoxicity represents a serious pitfall particularly when multiple doses are used to label cells. PMID- 14755514 TI - OL-protocadherin expression in the visual system of the chicken embryo. AB - The expression of OL-protocadherin, a homotypically binding cell adhesion molecule, was mapped in the visual system of the chicken embryo at intermediate to late stages of development (11-19 days of incubation). The expression was compared with that of four classic cadherins, described previously. OL protocadherin is expressed by the isthmooptic nucleus, its retinopetal projection, and possibly its retinal target neurons, the amacrine cells. Ganglion cells begin to express OL-protocadherin at relatively late stages of development. The layers of the optic tectum, the projection neurons in the stratum griseum centrale, and the tectofugal pathways show differential OL-protocadherin immunoreactivity. Several of the diencephalic target nuclei of the tectothalamic projection, such as the principal pretectal nucleus, subpretectal nucleus, and nucleus rotundus, contain distinct subregions or populations of neurons expressing OL-protocadherin. In these centers, the expression pattern of OL protocadherin differs from that of the four classic cadherins, though it shows partial overlap with them. Other retinorecipient and/or tectorecipient nuclei (ventral geniculate nucleus, lateral dorsolateral nucleus, superficial synencephalic nucleus, pretectal area, and griseum tectale) also show a differential immunoreactivity for OL-protocadherin and other cadherins. Some of these nuclei and the optic tectum display a similar sequence of cadherin expression from superficial to deep layers, in a pattern that may reflect mutual interconnections. This result indicates a partial conservation of cadherin expression across interconnected embryonic divisions, from the mesencephalon to the ventral thalamus. In conclusion, OL-protocadherin is a marker for specific functional gray matter structures and neural circuits in the chicken visual system. J. Comp. Neurol. 470:240-255, 2004. PMID- 14755515 TI - Characterization of neuropeptide Y Y2 and Y5 receptor expression in the mouse hypothalamus. AB - Neuropeptide Y (NPY) neurons abundantly innervate the hypothalamus, where NPY is involved in the regulation of a broad range of homeostatic functions. In the present work we studied NPY Y2 and Y5 receptor (R) gene expression in the mouse hypothalamus by using immunohistochemical detection of beta-galactosidase (beta gal), a gene reporter molecule for Y2R and Y5R in Y2R-knockout (KO) and Y5R-KO mice, respectively. With this approach, cells normally expressing Y2R or Y5R are immunopositive for beta-gal. In the hypothalamus of the Y2R-KO mouse, beta-gal immunoreactivity (-ir) was found in numerous neurons of the medial preoptic nucleus as well as in the lateral anterior, periventricular, dorsomedial, tuberal, perifornical, and arcuate nuclei. Most of the dopaminergic neurons in the A13 dorsal hypothalamic group were beta-gal positive, whereas other hypothalamic dopaminergic neurons rarely displayed beta-gal-ir. In the arcuate nucleus, most of the beta-gal-positive neurons expressed NPY, but colocalizations with beta-endorphin were also found; in the tuberal and perifornical nuclei, many beta-gal-positive neurons contained nitric oxide synthase. beta-Gal-ir was also found in other forebrain regions of the Y2R-KO mouse, including the amygdala, thalamic nuclei, hippocampal CA3 area, and cortex. In the hypothalamus of the Y5R KO mouse, beta-gal-positive neurons were found mainly in the arcuate nucleus and contained beta-endorphin. The present data show that Y2R and Y5R are expressed in distinct groups of hypothalamic neurons. High levels of Y2R expression in the preoptic nuclei suggest an involvement of Y2R in the regulation of reproductive behavior, whereas Y2R expression in the arcuate, dorsomedial, and perifornical nuclei may be relevant to feeding and body weight control. The finding that A13 dopaminergic neurons express Y2R suggests a new mechanism putatively involved in the central control of feeding, in which NPY can modulate dopamine secretion. The distribution of Y5R expression supports earlier evidence for involvement of this receptor in control of feeding and body weight via NPY's action on proopiomelanocortin-expressing neurons. J. Comp. Neurol. 470:256-265, 2004. PMID- 14755516 TI - Characterization of synaptogyrin 3 as a new synaptic vesicle protein. AB - Synaptogyrins comprise a family of tyrosine-phosphorylated proteins with two neuronal (synaptogyrins 1 and 3) and one ubiquitous (cellugyrin) isoform. Previous studies have indicated that synaptogyrins are involved in the regulation of neurotransmitter release. Synaptogyrin 1 is a synaptic vesicle protein; cellugyrin, by contrast, is absent from synaptic vesicles. In an effort to further characterize the synaptogyrin family, we studied the distribution of the synaptogyrin 3 protein in the nervous system. Subcellular fractionation and immunoprecipitation of synaptic vesicles from mouse brain showed that synaptogyrin 3 is associated with synaptic vesicles and that synaptogyrins 1 and 3 can reside on the same synaptic vesicle. Immunofluorescent staining of cultured hippocampal neurons confirmed the synaptic localization of synaptogyrin 3. Analysis of the relative distributions of synaptogyrins 1 and 3 in mouse brain revealed a more restricted expression pattern for synaptogyrin 3 compared to the ubiquitous distribution of synaptogyrin 1. Strong synaptogyrin 3 labeling was observed in the mossy fiber region of the hippocampus, substantia nigra pars reticulata, pallidum, and deep cerebellar nuclei. By comparison, the striatum and reticular and ventral posterolateral thalamic nuclei, which all showed synaptogyrin 1 labeling, contained significantly less synaptogyrin 3. Finally, we used in situ hybridization experiments to correlate synaptogyrin 3 mRNA in cell bodies with synaptogyrin 3 protein at synapses. Altogether, our data indicate that neuronal synaptogyrins are differentially expressed protein isoforms that may represent functionally distinct populations of synapses and/or synaptic vesicles. PMID- 14755517 TI - Fos expression in otolith-related brainstem neurons of postnatal rats following off-vertical axis rotation. AB - To determine the critical time of responsiveness of developing otolith organ related brainstem neurons and their distribution, Fos protein expression in response to off-vertical axis rotations (OVAR) was mapped in conscious Sprague Dawley rats from P5 to adulthood. OVAR was used to activate sequentially all utricular hair cells per 360 degrees revolution. We detected the coding of horizontal head positions in otolith organ-related neurons within the vestibular nucleus as early as P7. In the vestibular nuclear complex and its subgroups, the density of Fos-immunoreactive (Fos-ir) neurons increased steadily with age and reached the adult level by P21. In both labyrinthectomized rats subjected to OVAR and normal rats kept stationary, labeled neurons were found sporadically in the aforementioned brain regions in each age group, confirming that Fos labeling observed in neurons of normal experimental rats subjected to OVAR was due to otolith organ stimulation. Whereas OVAR-induced Fos-ir neurons were also first observed in vestibular-related brain areas, such as the prepositus hypoglossal nucleus, gigantocellular reticular nucleus, and locus coeruleus, of normal experimental rats at P7, those in the inferior olive were observed only from P14 onward. This indicates the unique maturation time of inferior olivary neurons in gravity-related spatial coding. In general, age-dependent increase in OVAR induced Fos-ir neurons was observed in brain areas that received otolith inputs. The locus coeruleus was exceptional in that prominent OVAR-induced Fos-ir neuronal number did not change with maturation, and this was well above the low but significant number of Fos-ir neurons in control preparations. Taken together, our results suggest that neuronal subpopulations within the developing network of the horizontal otolith system provide an anatomical basis for the postnatal development of otolith organ-related sensorimotor functions. J. Comp. Neurol. 470:282-296, 2004. PMID- 14755518 TI - Retinoic acid signaling in the brain marks formation of optic projections, maturation of the dorsal telencephalon, and function of limbic sites. AB - As retinoic acid (RA) is known to regulate the expression of many neuronal proteins, it is likely to influence overall development and function of the brain; few particulars, however, are available about its role in neurobiological contexts due mainly to problems in RA detection. To ask whether the function of RA in the rostral brain is concentrated in particular neurobiological systems, we compared sites of RA synthesis and actions, as detected by RA signaling in reporter mice, for embryonic and adult ages. We found that most sites of RA actions in the forebrain do not colocalize with RA synthesis, consistent with a dominant RA supply by diffusion and the circulation. The changing RA patterns distinguish preferentially two complex functional schemes. (1) Within the visual system when the first optic axons grow toward their targets, RA signaling delineates the topographical adjustment of the retinal map, which is encoded in the coordinates of the visual world, to central visual maps, which are formed in the segmental brain coordinates. (2) The second scheme begins early in forebrain morphogenesis as a distinction of the dorsal telencephalon. With progressing development, and in the adult, the RA patterns then focus on widely distributed structures, most of which belong to the limbic system. These are sites in which emotional perception is combined with higher cognitive processes and in which normal function requires ongoing remodeling of synaptic connections, indicating that the developmental role of RA in promotion of neuronal differentiation programs continues in the adult brain for highly flexible neural circuits. J. Comp. Neurol. 470:297-316, 2004. PMID- 14755519 TI - Dendritic morphology, local circuitry, and intrinsic electrophysiology of principal neurons in the entorhinal cortex of macaque monkeys. AB - Little is known about the neuroanatomical or electrophysiological properties of individual neurons in the primate entorhinal cortex. We have used intracellular recording and biocytin-labeling techniques in the entorhinal slice preparation from macaque monkeys to investigate the morphology and intrinsic electrophysiology of principal neurons. These neurons have previously been studied most extensively in rats. In monkeys, layer II neurons are usually stellate cells, as in rats, but they occasionally have a pyramidal shape. They tend to discharge trains, not bursts, of action potentials, and some display subthreshold membrane potential oscillations. Layer III neurons are pyramidal, and they do not appear to display membrane potential oscillations. The distribution of dendrites and of axon collaterals suggests that neurons in layers II and III are interconnected by a network of associational fibers. Layer V and VI neurons are pyramidal and tend to discharge trains of action potentials. The distribution of dendrites and axon collaterals suggests that there is an associative network of principal neurons in layers V and VI, and they also project axon collaterals toward superficial layers. Importantly, entorhinal cortical neurons in monkeys appear to exhibit significant differences from those in rats. Morphologically, neurons in monkey entorhinal layers II and III have more primary dendrites, more dendritic branches, and greater total dendritic length than in rats. Electrophysiologically, layer II neurons in monkeys exhibit less sag, and subthreshold oscillations are less robust and slower. Some monkey layer III neurons discharge bursts of action potentials that are not found in rats. The interspecies differences revealed by this study may influence information processing and pathophysiological processes in the primate entorhinal cortex. J. Comp. Neurol. 470:317-329, 2004. PMID- 14755520 TI - Distinct muscle targets do not vary in the developmental regulation of brain derived neurotrophic factor. J Comp Neurol 470:317-329,2004. AB - Developing neurons depend on many target-derived signals. One of these signals is the neurotrophin brain-derived neurotrophic factor (BDNF). Exogenous application of BDNF in vitro and in vivo rescues a population of lumbar motoneurons from programmed cell death. Given that BDNF does not rescue all motoneurons and that motoneurons differ in trophic factor receptor expression, subpopulations of motoneurons may have different sensitivities to the factor. These differences may be reflected in distinct target muscles specialized to produce different protein concentrations, or muscles may contain equal amounts of the factor and receptor expression determines motoneuron responsiveness. By using a sensitive electrochemiluminescent immunoassay (ECLIA), we measured normal developmental changes in BDNF protein concentration in anatomically and functionally distinct chick embryonic thigh muscles from E6 to E18. We found that there were no significant differences in BDNF protein concentration between muscles classified according to function (fast vs. slow) or anatomical position (flexor vs. extensor) and that the quantity of BDNF in the target did not appear to be activity dependent. These results suggest that, during development, the differences in the response of motoneurons to BDNF are not due to the anatomical or functional diversity of muscle targets. J. Comp. Neurol. 470:330-337, 2004. PMID- 14755521 TI - Connexin35/36 gap junction proteins are expressed in photoreceptors of the tiger salamander retina. AB - Photoreceptors in the vertebrate retina are electrically coupled with one another. Such coupling plays important roles in visual information processing. Physiological properties of rod-rod and rod-cone coupling have been best studied in the salamander retina, yet the cellular and molecular basis of these electrical synapses has not been established. Recently, connexin35/36 (Cx35/36) gap junction proteins were found to be highly expressed in brain and retina, suggesting that it may mediate photoreceptor coupling. To test this idea, we examined the cellular distribution of Cx35/36 in the salamander retina. Western blot analysis showed the expression of Cx35/36 proteins, and confocal microscopy revealed characteristic punctate Cx35/36 immunoreactivity in both synaptic layers. In addition, Cx35/36-positive plaques were detected in the outer nuclear layer (ONL) between neighboring rods, and these plaques outlined the mosaic of the rod network at a level distal to the external limiting membrane. Moreover, although Cx35/36 plaques were detected between some cones and their adjacent rods, the number and size of these plaques was smaller, and their staining intensity was diminished compared with the plaques between adjacent rods. Furthermore, Lucifer yellow injection together with confocal microscopy revealed that Cx35/36-puncta were colocalized with finlike structures of rod cell membrane, with the ultrastructure of gap junctions between paired rod fins having been found by electron microscopy. Therefore, our findings demonstrate that Cx35/36 expression in photoreceptors is primarily located between rods and to a lesser extent between rods and cones, suggesting that Cx35/36 may participate in electrical coupling between rods and between rods and cones in the salamander retina. PMID- 14755523 TI - Evolutionary changes in the olfactory projection neuron pathways of eumalacostracan crustaceans. AB - Output from the olfactory lobe (primary olfactory center) of eumalacostracan crustaceans is transmitted to the medulla terminalis (MT) and hemiellipsoid body (HB) in the lateral protocerebrum (higher order center) by a large population of projection neurons. In eureptantian crustaceans (lobsters, crayfish, and crabs), these projection neurons also form the output pathway from an additional neuropil, the accessory lobe (higher order center), which appears to have arisen de novo in these animals. In a previous study of lobsters and crayfish we showed that whereas projection neurons innervating the olfactory lobe project primarily to the MT, those innervating the accessory lobe project exclusively to the HB (Sullivan and Beltz [ 2001a] J. Comp. Neurol. 441:9-22). In the present study, we used focal dye injections to examine the olfactory projection neuron pathways of representatives of four eumalacostracan taxa (Stomatopoda, Dendrobranchiata, Caridea, and Stenopodidea) that diverged from the eureptantian line prior to the appearance of the accessory lobe. These experiments were undertaken both to examine the evolution of the olfactory pathway in the Eumalacostraca and to provide insights into the changes in this pathway that accompanied the appearance of the accessory lobe. The innervation patterns of the olfactory projection neurons of the species examined were found to differ markedly, varying from that observed in the most basal taxon examined (Stomatopoda), in which the neurons primarily project to the MT, to those observed in the two highest taxa examined (Caridea and Stenopodidea), in which they primarily target the HB. These results suggest that substantial changes in the relative importance of the MT and HB within the olfactory pathway have occurred during the evolution of the Eumalacostraca. PMID- 14755522 TI - Sema3A regulates the timing of target contact by cranial sensory axons. AB - The trigeminal ganglion provides the somatosensory innervation for the anterior rat tongue. At early embryonic stages (embryonic day [E] 12-13) pre-tongue explants repel trigeminal axon outgrowth, and this is mediated by Sema3A (Rochlin and Farbman [1998] J. Neurosci. 18:6840-6852; Rochlin et al. [2000] J. Comp. Neurol. 422:579-593). Despite a decrease in repulsion by E14 and older tongue explants, Sema3A mRNA persists throughout the dorsal epithelium through E18, after axons have begun to penetrate papilla epithelium. We investigated the hypothesis that Sema3A continues to act as a repellent and that subpopulations of trigeminal axons that penetrate the epithelium become unresponsive to Sema3A. Sema3A repelled trigeminal axons in vitro regardless of the neurotrophic factor used to stimulate axon outgrowth, but the minimum level of Sema3A required to repel depended on the neurotrophic factor. Thus, in vitro, trigeminal axons are repelled by Sema3A when they would be penetrating the Sema3A-mRNA rich epithelium in vivo. Whereas dorsal epithelium on tongue explants dissected at stages preceding target contact (E15) repelled trigeminal axons in vitro, explants dissected at later stages (E18), after axons would have penetrated the epithelium in vivo, were not repellent. To determine whether Sema3A prevents premature target penetration in vivo, we assessed the timing of target contact by sensory axons in Sema3A-/minus; and +/+ mice. Contact of the epithelium occurs prematurely in Sema3A-/minus; mice, but not penetration. Taken together, our data imply that Sema3A acts as a short-range repellent that regulates the timing of target contact by trigeminal axons. PMID- 14755524 TI - Re-evaluation of the afferent connections of the pituitary in the weakly electric fish Apteronotus leptorhynchus: an in vitro tract-tracing study. AB - The pituitary plays a key role in the interaction between the brain and the endocrine system. We re-examined the afferent connections of the pituitary in the weakly electric fish Apteronotus leptorhynchus using the in vitro application of dextran-tetramethylrhodamine to the pituitary. The resultant retrograde labeling was analyzed. Application of the tracer to the rostral part, but not the caudal part, of the pituitary labels hypothalamic cells in the anterior division of the periventricular nucleus, the suprachiasmatic nucleus, and the nucleus tuberis lateralis pars anterior. Application of the tracer to either the rostral or caudal parts of the pituitary labels hypothalamic cells in the posterior division of the periventricular nucleus (RPPp), the nucleus hypothalamus caudalis (Hc), the nucleus hypothalamus anterioris, the ventral hypothalamic nucleus, and the central nucleus of the inferior lobe. Furthermore, cells in the rostral two thirds of the brainstem reticular formation (RF) project to the entire rostrocaudal extent of the pituitary. The largest projections to the pituitary are from Hc, PPp, and RF. Of the cells in Hc that project to the pituitary, almost all (96%) are small and the remainder are medium-sized. Of the cells in PPp that project to the pituitary, about half are small or medium-sized (44% and 56%, respectively). In Hc and PPp, about one-third to one-half of the cells that project to the pituitary are markedly elongated. The cells in RF that project to the pituitary are small (4%), medium-sized (89%), or large (7%) and about four fifths of these cells are markedly elongated. With regard to the RF projections, the pituitary may receive copies of motor instructions and sensory information supplied by collaterals of the descending and ascending projection systems of RF cells. Thus, the ongoing motor activity of the animal and the ensuing sensory feedback from this activity could directly influence the pituitary. PMID- 14755525 TI - Response of olfactory ensheathing cells to the degeneration and regeneration of the peripheral olfactory system and the involvement of the neuregulins. AB - In this study we examined the proliferative response of olfactory ensheathing cells (OECs) to olfactory receptor neuron injury induced by zinc sulfate (ZnSO4) irrigation and related the response of OECs within the peripheral system to the inflammatory response induced by injury and the expression profile of neuregulins. After ZnSO4 treatment, degeneration in the epithelium is reproducible and rapid, with regeneration following after 4 days, and is morphologically complete by 5 weeks. Changes in the olfactory bulb are less dramatic, although degeneration of both the outer and the glomerular layers occurred. Treatment also induced a marked inflammatory response in both the epithelium and the bulb. Unlike Schwann cell changes associated with Wallerian degeneration, OECs did not proliferate or obviously migrate within the olfactory system in response to axonal loss, suggesting that the new nerves generated from the epithelium regrow back through conduits already formed by the glia. Expression of neuregulin 1alpha was maintained in the nerve by OECs, and changes in neuregulin 1 mRNA and erbB2 mRNA expression were detected, indicating that these growth factors may play a role in the regeneration of the peripheral olfactory system but not in OEC proliferation. PMID- 14755526 TI - Alterations in calretinin immunostaining in the ferret superior olivary complex after cochlear ablation. AB - In this study, we used image analysis to assess changes in calretinin immunoreactivity in the lateral (LSO) and medial (MSO) superior olivary nuclei in ferrets 2 months after unilateral cochlear ablations at 30-40 days of age, soon after hearing onset. These two nuclei are the first significant sites of binaural convergence in the ascending auditory system, and both receive direct projections from the deafferented cochlear nucleus. Cochlear ablation results in a decrease in the overall level of calretinin immunostaining within the LSO ipsilaterally compared with the contralateral side and with control animals and within the MSO bilaterally compared with control ferrets. In addition, the level of calretinin immunostaining ipsilaterally within neurons in the LSO was significantly less in cochlear ablated than control animals. In contrast, there was no effect of cochlear ablation on the level of calretinin immunostaining within neurons either in the contralateral LSO or in the MSO. These results are consistent with a downregulation in calretinin within the neuropil of MSO bilaterally and LSO ipsilaterally, as well as a downregulation in calretinin within somata in the ipsilateral LSO as a result of unilateral cochlear ablation soon after hearing onset. Thus, cochlear-driven activity appears to affect calcium binding protein levels in both neuropil and neurons within the superior olivary complex. PMID- 14755527 TI - In vitro analysis of mechanisms underlying age-dependent failure of axon regeneration. AB - Severed axons of the inferior colliculus (IC) commissure can regenerate across a lesion in organotypic cultures from postnatal day (P) 6 gerbils, but this regenerative capacity is lost by P12 (Hafidi et al. [ 1995] J Neurosci 15:1298 1307, [1999] J Neurobiol 41:267-280). In the present study, we examined the mechanisms underlying this age-dependent failure of axons to regenerate. In P6 P12 heterochronic cultures, the P12 axons failed to cross the lesion site and project to the contralateral P6 IC lobe. In contrast, axons originating from the P6 lobe could regenerate through the lesion and invade the contralateral P12 IC lobe. To determine whether this age-dependent change in regenerative capacity can develop in organotypic cultures, IC slices with an intact commissure were obtained from P6 animals, grown in vitro for 6 days, and then lesioned at the commissure. In these slices, axon regeneration failure was similar to that observed in normal P12 tissue. Several in vitro treatments enhanced axon regeneration: removal of the entire midline region, inhibition of protein synthesis at the lesion site, and exposure to ABC chondroitinase. Furthermore, when the injured commissural axons were provided with a carpet of C6-R cells (a radial glia-like cell line), significantly more axons projected to the contralateral lobe of the IC. Taken together, these results suggest that the maturation of nonneuronal cells within the lesion site lead to failed axon regeneration in mature animals, and show that ameliorative strategies can be evaluated in vitro. PMID- 14755528 TI - Activity-dependent regulation of the potassium channel subunits Kv1.1 and Kv3.1. AB - Afferent activity, especially in young animals, can have profound influences on postsynaptic neuronal structure, function and metabolic processes. Most studies evaluating activity regulation of cellular components have examined the expression of ubiquitous cellular proteins as opposed to molecules that are specialized in the neurons of interest. Here we consider the regulation of two proteins (voltage-gated potassium channel subunits Kv1.1 and Kv3.1) that auditory brainstem neurons in birds and mammals express at uniquely high levels. Unilateral removal of the avian cochlea leads to rapid and dramatic reduction in the expression of both proteins in the nucleus magnocellularis (NM; a division of the avian cochlear nucleus) neurons as detected by immunocytochemistry. Uniform downregulation of Kv1.1 was reliable by 3 hours after cochlea removal, was sustained through 96 hours, and returned to control levels in the surviving neurons by 2 weeks. The activity-dependent changes in Kv3.1 appear to be bimodal and are more transient, being observed at 3 hours after cochlea removal and recovering to control levels within 24 hours. We also explored the functional properties of Kv1.1 in NM neurons deprived of auditory input for 24 hours by whole-cell recordings. Low-threshold potassium currents in deprived NM neurons were not significantly different from control neurons in their amplitude or sensitivity to dendrotoxin-I, a selective K+ channel antagonist. We conclude that the highly specialized abundant expression of Kv1.1 and 3.1 channel subunits is not permanently regulated by synaptic activity and that changes in overall protein levels do not predict membrane pools. PMID- 14755529 TI - Multipotency of purified, transplanted globose basal cells in olfactory epithelium. AB - By comparison with the rest of the nervous system, the olfactory epithelium has an unparalleled ability to renew and repair itself throughout life. However, the identity and capacity of the various types of progenitor cells that underlie that ability are not well established. We used selective isolation, transplantation, and engraftment of various types of marker-labeled cells into the epithelium of methyl bromide-lesioned, unmarked host mice to dissect progenitor cell capacity. Globose basal cells were purified from other potential progenitors using the monoclonal antibody GBC-2 (GBC, globose basal cell) and fluorescence activated cell sorting. Transplanted globose basal cells engraft and, in aggregate, give rise to globose basal cells, neurons, sustentacular cells, and several other kinds of non-neuronal cells. Individual clones, derived from single engrafted globose basal cells, can consist of a mixture of neurons and non-neuronal cells, only neurons, or only non-neuronal cells. Neurons that arise after transplantation mature to the point of expressing odorant receptors and olfactory marker protein and of projecting axons to the olfactory bulb. In contrast, other kinds of epithelial cells are neither neurogenic nor multipotent. For example, sustentacular and duct cells give rise only to themselves after transplantation. Furthermore, horizontal basal cells do not engraft in mice, in which the endogenous population is spared after lesion. Thus, some subtype(s) of GBC is a multipotent progenitor cell, whose multipotency is activated after destruction of both neurons and non-neuronal cells. The results suggest that progenitor cell transplantation may prove useful as a therapeutic modality as well as an analytical tool. PMID- 14755530 TI - Altered epithelial density and expansion of bulbar projections of a discrete HSP70 immunoreactive subpopulation of rat olfactory receptor neurons in reconstituting olfactory epithelium following exposure to methyl bromide. AB - A previously described subpopulation of rat olfactory receptor neurons, the 2A4(+)ORNs, is 1) distinguished by intense constitutive cytoplasmic immunoreactivity to antibodies to the 70-kD heat shock protein (HSP70); 2) occurs sparsely but consistently through ventral and lateral olfactory epithelium (OE); and 3) projects to just two to three consistently located glomeruli in each olfactory bulb (OB) (Carr et al. [1994] J Comp Neurol 348:150-160). Immunoreactivity appears not to be stress-related. To examine the persistence of these features following destruction and reconstitution of the OE, rats were subjected to methyl bromide-induced OE lesion (Schwob et al. [1995] J Comp Neurol 59:15-37; Schwob et al. [1999] J Comp Neurol 412:439-457] and their OE and OBs examined with antibodies to HSP70 6-10.5 weeks postlesion. Lesioned OE showed significantly increased 2A4(+)ORN densities but no alteration of 2A4(+)ORN zonal distribution. The OBs of lesioned animals showed marked expansions of 2A4(+)ORN bulbar projections, with 2-15-fold increases in numbers of glomeruli showing 2A4(+)axons, and projection expansions were greater in animals maintained on chronic food restriction prior to lesioning. Examination of archival 5-month post MeBr lesion material indicates that altered projection patterns are maintained. PMID- 14755531 TI - Synaptic connectivity of the diffuse bipolar cell type DB6 in the inner plexiform layer of primate retina. AB - Diffuse bipolar cells in primate retina receive synaptic input from multiple cones and provide output to ganglion cells. Diffuse bipolar cells can be subdivided into six types (DB1-DB6) according to the stratification of their axon terminals in the inner plexiform layer, but their synaptic connectivity in the inner plexiform layer is not well understood. Here the stratification and synaptic connectivity of DB6 axon terminals were studied in the retinae of New World (marmoset) and Old World (macaque) monkeys. Immunohistochemical markers were applied to retinal sections. The sections were analyzed by confocal and deconvolution light microscopy as well as electron microscopy. The DB6 cells were identified with antibodies against CD15; rod bipolar cells were identified with antibodies against protein kinase Calpha (PKCalpha); and AII amacrine cells were identified with antibodies against calretinin. The axons of DB6 and rod bipolar cells occupy distinct regions in stratum 5 of the inner plexiform layer. The distal processes of calretinin-labeled AII cells are usually closely associated with rod bipolar axons but sometimes also with DB6 axons. Pre-embedding immunoelectron microscopy showed that the vast majority (over 86%) of the synaptic output of DB6 cells is onto amacrine cell processes, whereas less than 14% goes to ganglion cell processes. In double-labeled preparations DB6 axons occasionally made output onto calretinin-labeled amacrine processes. Thus it is possible that AII cells receive some input from DB6 cells. PMID- 14755532 TI - Effects of two years of estrogen loss or replacement on nucleus basalis cholinergic neurons and cholinergic fibers to the dorsolateral prefrontal and inferior parietal cortex of monkeys. AB - The present study examined the long-term (2 years) effects of estrogen loss or estrogen replacement therapy (ERT) on cholinergic neurons in the nucleus basalis of Meynert and on cholinergic fibers in the prefrontal and parietal cortex of adult female cynomolgus monkeys. Cholinergic fiber density in layer II of the prefrontal cortex was decreased in monkeys who were ovariectomized and treated with placebo for 2 years. In contrast, ovariectomized monkeys receiving ERT for 2 years had fiber densities that were comparable to those of intact controls. No differences in parietal cholinergic fiber density or nucleus basalis cholinergic neuron number or volume were found among intact, ovariectomized, or ERT monkeys. Our results suggest that ERT is effective in preventing region-specific changes in cortical cholinergic fibers that result from the loss of circulating ovarian hormones. These modest but appreciable effects on cholinergic neurobiology following long-term estrogen loss and ERT may contribute to changes in visuospatial attention function that is mediated by the prefrontal cortex. PMID- 14755534 TI - Expression of androgen receptor mRNA in zebra finch song system: developmental regulation by estrogen. AB - By using improved methods for in situ hybridization to detect expression of androgen receptor (AR) mRNA, the distribution of expression was mapped in the adult male zebra finch brain. In the neural song circuit, robust expression was found in area X of the lobus parolfactorius (LPO) as well as in other song regions previously reported. Expression was also found in many areas of the hypothalamus and dorsal thalamic nuclei, nucleus intercollicularis and ventricular areas of the midbrain, cerebellar Purkinje and granule cells, the hyperstriatum, medial neostriatum, medial LPO, and archistriatum. In juvenile males, AR mRNA expression was first detected in nucleus high vocal center (HVC) at posthatch day 9 (P9), in area X at P9-P11, and in the region of the robust nucleus (RA) in the medial archistriatum by P7. Estrogen treatment of hatchling females caused an increase in the expression of AR mRNA in HVC and area X by P11, whereas treatment of hatchling males with the aromatase inhibitor fadrozole decreased the expression of AR mRNA at P11. The present results indicate that masculine development of AR expression begins in area X and HVC before they are thought to be synaptically connected, suggesting that different song nuclei initiate sexual differentiation independently of transsynaptic masculinizing influences. The present results suggest that estrogen is necessary for full masculine AR expression in the song system and that the estrogenic regulation of AR contributes to subsequent differential actions of androgen in male and female song nuclei. PMID- 14755533 TI - Presynaptic N-methyl-D-aspartate receptor expression is increased by estrogen in an aromatase-rich area of the songbird hippocampus. AB - The vertebrate hippocampus (HP) is sensitive to estrogens, in part via effects on N-methyl-D-aspartate (NMDA)-type glutamate receptors (NR). Although the precise mechanism of this interaction is unclear, it constitutes a key interface in the plasticity of the adult vertebrate HP. The songbird HP expresses high levels of aromatase (estrogen synthase), suggesting that locally generated steroid may affect excitatory pathways. By using light, confocal, and electron microscopy with antibodies that specifically recognize aromatase and NR, we have 1) mapped their distribution in the zebra finch brain, 2) documented their coexpression in HP neurons, 3) studied the ultrastructure of NR-expressing cells in the HP, and 4) tested the influence of estrogen on the cellular and subcellular characteristics of NR-positive HP neurons. Aromatase and NR are coexpressed in HP neurons. NRs are detectable in presynaptic boutons of the songbird HP in addition to postsynaptic loci. Treatment with estrogen increased the somal size and innervation of NR-positive neurons and the frequency of presynaptic NR. Autoreception of excitatory neurotransmission via presynaptic NR may promote the strengthening of activity-dependent, excitatory synapses, thereby enhancing learning. NR-mediated autoreception may underlie estrogenic enhancement of HP structural and functional plasticity. PMID- 14755535 TI - Morphology and connections of the abdominal accessory neurons of the crayfish Cherax destructor. AB - Associated with the abdominal muscle receptor organs of crayfish are accessory neurons that inhibit the activity of the stretch receptors. Cobalt infusion into their cut axons reveals four accessory somata associated with each hemiganglion in the abdomen of the crayfish Cherax destructor. These conform to the pattern described previously for these neurons: The cell bodies are in the ganglion posterior to the one from which they exit. We recorded intracellularly from the largest accessory neurons, Acc-1 and Acc-2, and stained them with intracellular dye to establish unambiguously the characteristics defining their identity and structure. We describe their branching patterns in the ganglion of origin and the ganglion of exit. This morphological information permitted us to distinguish all four accessory neurons in preparations with dye infused through their cut axons, and we propose a revised, unambiguous nomenclature for the two smaller ones. Our intracelluar recordings allowed us to reexamine the physiological relationships of Acc-1 and Acc-2, the only accessory neurons for which there are data in the literature. In general, the connections and inputs described in previous studies were substantiated, although there has clearly been confusion between the two, and they differ in a number of significant ways. We found that they are seldom active together, have different firing patterns, and may operate with different clusters of extensor and flexor motorneurons. The results illustrate the level at which the accessory neurons operate within the abdominal control system but do not distinguish between competing hypotheses concerning their role in behavior. The data are consistent with the view that accessory neurons assist in timing between adjacent segments. PMID- 14755536 TI - Distribution of corticotropin-releasing factor-immunoreactive neurons in the central nervous system of the domestic chicken and Japanese quail. AB - In birds, as in mammals, corticotropin-releasing factor (CRF) is present in a number of extrahypothalamic brain regions, indicating that CRF may play a role in physiological and behavioral responses other than the control of adrenocorticotropin hormone release by the pituitary. To provide a foundation for investigation of the roles of CRF in the control of avian behavior, the distribution of CRF immunoreactivity was determined throughout the central nervous system of the domestic chicken (Gallus domesticus) and Japanese quail (Coturnix japonica). The distribution of CRF-immunoreactive (-ir) perikarya and fibers in the chicken and quail brain was found to be more extensive than previously reported, notably in the telencephalon. Numerous CRF-ir perikarya and fibers were present in the hyperstriatum, hippocampus, neostriatum, lobus parolfactorius, and archistriatum, as well as in the nucleus taeniae, nucleus accumbens, and bed nucleus of the stria terminalis, which exhibited the strongest immunolabeling in the telencephalon. The presence of dense populations of CRF-ir perikarya in the medial lobus parolfactorius, nucleus of the stria terminalis, and paleostriatum ventrale, apparently giving rise to CRF-ir projections to the mesencephalic reticular formation, the parabrachial/pericerulear region, and the dorsal vagal complex, suggests that these telencephalic areas may constitute part of the avian "central extended amygdala." These results have important implications for understanding the role of extrahypothalamic CRF systems in emotional responses in birds. PMID- 14755537 TI - Axonal projections from the parasubthalamic nucleus. AB - The parasubthalamic nucleus (PSTN) is a differentiation of the lateral hypothalamic area, characterized by a distinct population of neurons expressing beta-preprotachykinin (beta-PPT) mRNA. The axonal projections from the PSTN have been analyzed with the PHAL anterograde tract tracing method in rats. The results indicate that the cell group is distinguished by massive projections to parasympathetic preganglionic neurons in the brainstem (especially in the salivatory nuclei and dorsal motor nucleus of the vagus nerve) and to parts of the parabrachial nucleus and nucleus of the solitary tract that relay viscerosensory and gustatory information. In addition, the PSTN projects to cortical parts of the cerebral hemisphere (infralimbic, agranular insular, postpiriform transition and lateral entorhinal areas, and posterior basolateral amygdalar nucleus)-directly and also indirectly via thalamic feedback loops involving the paraventricular and mediodorsal nuclei-and to nuclear parts of the cerebral hemisphere (central amygdalar nucleus, striatal fundus, rhomboid nucleus of the bed nuclei of the stria terminalis, and substantia innominata). The PSTN is thus positioned to influence directly many cerebral hemisphere and hindbrain components of the central parasympathetic control network that is active, for example, during feeding behavior and cardiovascular regulation. PMID- 14755538 TI - Kinase suppressor of RAS (KSR) amplifies the differentiation signal provided by low concentrations 1,25-dihydroxyvitamin D3. AB - The activity of kinase suppressor of ras (KSR), a kinase or a molecular scaffold upstream from Raf-1, is involved in the MEK/ERK MAP kinase cascade which can signal cell growth, survival, or differentiation, depending on the cellular context. We provide evidence here that KSR is upregulated in HL60 cells undergoing differentiation induced by low (0.3-3 nM) concentrations of 1,25 dihydroxyvitamin D(3) (1,25D(3)), and an antisense oligo (AS), but not a sense oligo, to KSR inhibits this differentiation. The inhibition of differentiation by AS-KSR oligo was less apparent when the concentration of 1,25D(3) was increased, suggesting that at the higher concentrations of 1,25D(3) KSR is not essential for the signaling of the differentiated phenotype. The reduced differentiation of HL60 cells exposed to AS-KSR was paralleled by reduced phosphorylation of Raf-1 Ser 259, and of p90RSK, used here as read-out for MAPK cascade activity. Conversely, ectopic expression of Flag-tagged wild type KSR potentiated the differentiation-inducing effects of low concentrations of 1,25D(3). Additional data suggest that the kinase activity of KSR is required for these effects, as transfection of a kinase inactive KSR construct did not significantly increase the 1,25D(3)-induced differentiation. Enzyme assays performed with KSR immunoprecipitated from 1,25D(3)-treated cells showed kinase activity when recombinant Raf-1 was used as the substrate, but not when the 1,25D(3)-treated cells were pretreated with AS-KSR oligos. Taken together, these data suggest that KSR participates in signaling of monocytic differentiation by augmenting the strength of the signal transmitted through Raf-1 to downstream targets. PMID- 14755539 TI - New basic discoveries and frontiers in diagnosis, prognosis and prediction of response to therapy in human thyroid, urinary bladder, and prostate tumors. AB - The fourth edition of this workshop mainly focused on three different human oncotypes, which included thyroid, urinary bladder, and prostate tumors as clinical models to gain new basic knowledge on tumor diagnosis, prognosis, and treatment. At the previous editions (Giordano et al., 2000, J Cell Physiol 183:284-287; Giordano et al., 2001, J Cell Physiol 188:274-280; Giordano et al., 2002, J Cell Physiol 191:362-365), leaders in the fields of pathology, clinical oncology, and basic research presented and discussed the most recent and prevalent findings in such neoplasms from a basic and clinical perspective. A concept that has been widely proposed is that the analysis of intrinsic biological factors displayed by primary tumors may be a valid method for diagnosing different neoplasias and for measuring both their aggressiveness and response to therapy. To date, however, no single prognostic factor, such as oncogenes, suppressor genes, or genes involved in the control of the cell cycle and/or apoptosis has yet proven to be potent enough to be used in clinical practice as a prognostic and predictive factor. The new possibility to simultaneously analyze the expression of the complete repertoire of human genes and a large number of proteins could offer a new scenario in tumor classification, allowing for the formulation of a list of genes able to define a "signature" of tumor outcome. Moreover, starting from data obtained from biomolecular tumor analyses, it has been demonstrated that with this approach, it is also possible to design future therapeutic strategies. PMID- 14755540 TI - Normal skin wound and hypertrophic scar myofibroblasts have differential responses to apoptotic inductors. AB - During wound healing, myofibroblasts play a central role in matrix formation and wound contraction. At the end of healing, there is evidence that myofibroblasts disappear via apoptotic pathways. Hypertrophic scars are a fibroproliferative disorder that leads to considerable morbidity. It has been postulated that a defect in myofibroblast apoptosis could be responsible for the pathological scar formation, but no evidence exists. We have isolated and cultured human normal wound (Wmyo) and hypertrophic scar (Hmyo) myofibroblasts and compared their basal apoptotic rates and their sensitivity to serum starvation and Fas antibody induced apoptosis to that obtained for dermal fibroblasts (Fb). A higher rate of apoptosis as evidenced by morphological criteria and a propidium iodide assay was observed for Wmyo in comparison to Fb and Hmyo. These results came along with a low level of the anti-apoptotic proteins Bcl-2 and Bclx(L) in Wmyo, whereas there was an increase in the level of the pro-apoptotic molecule Bax when compared to the results obtained for Fb and Hmyo. Hmyo showed a higher level of Bcl-2 compared to Fb but no difference in the Bax or Bclx(L) level. After serum starvation, Wmyo revealed an increased apoptotic rate, whereas Hmyo and Fb did not show any difference. Anti-Fas treatment did not modify the levels of apoptosis but strongly increased the cell growth of Hmyo as compared to Wmyo. This is the first study presenting a broad vision of the apoptotic sensitivity of normal and pathological myofibroblasts. These results confirmed the hypothesis of defects in apoptosis and growth during pathological scar formation impeding myofibroblast disappearance at the end of healing. PMID- 14755541 TI - Simultaneous activation of several second messengers in hypoxia-induced hyperpermeability of brain derived endothelial cells. AB - In vivo, ischemia is known to damage the blood-brain barrier (BBB) leading to the development of vasogenic brain edema. Hypoxia-induced vascular endothelial growth factor (VEGF) has been shown to be a key regulator of these permeability changes. However, the signaling pathways that underlie VEGF-induced hyperpermeability are incompletely understood. In this study, we demonstrate that hypoxia- and VEGF induced permeability changes depend on activation of phospholipase Cgamma (PLCgamma), phosphatidylinositol 3-kinase/Akt (PI3-K/Akt), and protein kinase G (PKG). Inhibition of mitogen-activated protein kinases (MAPK) and of the protein kinase C (PKC) did not affect permeability at all. Paralleling hypoxia- and VEGF induced permeability changes, localization of the tight junction proteins occludin, zonula occludens-1 (ZO-1), and ZO-2 along the cell membrane changed from a continuous to a more discontinuous expression pattern during hypoxia. In particular, localization of ZO-1 and ZO-2 expression moved from the cell membrane to the cytoplasm and nucleus whereas occludin expression remained at the cell membrane. Inhibition of PLCgamma, PI3-kinase, and PKG abolished these hypoxia induced changes. These findings demonstrate that hypoxia and VEGF induce permeability through rearrangement of endothelial junctional proteins which involves activation of the PLCgamma and PI3-K/AKT pathway leading to the activation of PKG. PMID- 14755542 TI - Different roles of p38 MAPK and ERK in STI571-induced multi-lineage differentiation of K562 cells. AB - STI571 is a specific tyrosine kinase inhibitor of Abl kinase. It was previously reported that STI571 induced hemoglobin synthesis in the chronic myelogenous leukemia (CML) cell line K562. However, its mechanisms remain unknown. In this study, we demonstrated that STI571 induced the phosphorylation of p38 mitogen activated protein kinase (MAPK) and dephosphorylation of extracellular signal regulated kinase (ERK) in K562 cells. In contrast, the phosphorylation of c-Jun N terminal kinases (JNK) in K562 cells was not altered by STI571. We also found that STI571 induced all the myeloid (CD11b, CD13), megakaryocytic (CD41a, CD42), and erythroid (glycophorin-A) markers on K562 cells. A p38 MAPK-specific inhibitor, SB203580, inhibited the STI571-induced multi-lineage differentiation of K562 cells, indicating that p38 MAPK is crucial for this differentiation. In contrast, SB203580 did not overcome the inhibitory effect for proliferation of K562 cells, indicating that p38 MAPK activation by STI571 does not affect cell numbers. Among the hematopoietic transcription factors, the expression level of c myb mRNA was clearly downregulated after incubation with STI571 in K562 cells. STI571-induced downregulation of c-myb mRNA was prevented by the pretreatment of K562 cells by SB203580. Our data provides insights into how p38 MAPK and ERK pathways are involved in STI571-induced differentiation of K562 cells. PMID- 14755543 TI - Characterization of Gicerin/MUC18/CD146 in the rat nervous system. AB - Gicerin is a cell adhesion molecule of an immunoglobulin (Ig) superfamily isolated from a chicken. It shows homophilic and heterophilic binding activities and has two isoforms. s-Gicerin which has small cytoplasmic domain and the same extracellular domain as l-gicerin shows stronger cell adhesion activity. In the chick nervous system, gicerin expression is only observed in the developmental stage when neurons extend neurites and migrate. In other tissues, gicerin participates in the tissue regeneration or oncogenesis. In this report, we identified two isoforms of rat gicerin corresponding to chicken and we concluded that gicerin is a homologue of human CD146/MUC18/MCAM. Next we generated antibody to characterize a rat gicerin in the nervous system. Gicerin is expressed in the hippocampal cells, Purkinje cells, and sensory neurons of a spinal chord of an adult rat, while expressed most abundantly in the lung. In addition to this, its expression in the hippocampus was increased by electroconvulsive shock, suggesting some role in the mature nervous system. And we also showed neurite promotion activity of gicerin from hippocampal neurons. PMID- 14755544 TI - Recruitment and proliferation of T lymphocytes is supported by IFNgamma- and TNFalpha-activated human osteoblasts: Involvement of CD54 (ICAM-1) and CD106 (VCAM-1) adhesion molecules and CXCR3 chemokine receptor. AB - The mechanism by which osteoblasts (OB) interact and modulate the phenotype and proliferation of T lymphocytes during inflammation is not well known. The effects of two regulatory cytokines, TNFalpha and IFNgamma, on the expression of CD54 (ICAM-1) and CD106 (VCAM-1) adhesion molecules and the CXCR3 ligands (CXCL9, CXCL10, CXCL11), were assessed in a primary culture of human OB by real-time PCR, flow cytometry, and immunohistochemistry. In addition, we functionally evaluated the recruitment and proliferation of T lymphocytes grown with resting or stimulated OB. According to the present data IFNgamma, either alone or in combination with TNFalpha, significantly up-regulates the expression of CD54 and CD106 and induces the expression and release of CXCL9, CXCL10, CXCL11 in OB. The supernatant of TNFalpha- and IFNgamma-activated OB induces the recruitment of T lymphocytes more significantly than stimulation by CXCR3 ligands. T lymphocyte proliferation is significantly enhanced by direct contact with TNFalpha- and IFNgamma-activated OB or by incubation with the supernatant of TNFalpha- and IFNgamma-activated OB. Blocking experiments with anti-CD11a, anti-CD49d, anti CXCR3, and Bordetella pertussis toxin demonstrate that adhesion molecules and the CXCR3 chemokine receptor play a key role in the proliferation of T lymphocytes. The present study demonstrates the involvement of adhesion molecules (CD11a and CD49d) and chemokine receptor (CXCR3) in the mechanism by which OB recruit, interact, and modulate T lymphocyte proliferation under inflammatory conditions. PMID- 14755545 TI - ERK1/2 and JNKs, but not p38 kinase, are involved in reactive oxygen species mediated induction of osteopontin gene expression by angiotensin II and interleukin-1beta in adult rat cardiac fibroblasts. AB - Osteopontin (OPN), also called cytokine Eta-1, expressed in the myocardium co incident with heart failure plays an important role in post myocardial infarction (MI) remodeling by promoting collagen synthesis and accumulation. Angiotensin II (Ang II) and inflammatory cytokines are increased in the heart following MI. We studied the involvement of mitogen-activated protein kinases (ERK1/2, JNKs, p38 kinase) and reactive oxygen species (ROS) in Ang II- and cytokine-induced OPN gene expression in adult rat cardiac fibroblasts. Ang II alone increased OPN mRNA (3.3 +/- 0.3-folds; P < 0.05; n = 7), while interleukin-1beta (IL-1beta), tumor necrosis factor (TNF-alpha), and interferon-gamma (IFN-gamma) had no effect. A combination of Ang II with IL-1beta or TNF-alpha, not IFN-gamma, increased OPN mRNA more than Ang II alone. Nitric oxide donor, S-nitrosoacetylpenicillamine (SNAP), alone or in combination with Ang II had no effect. Diphenylene iodonium (DPI), inhibitor of NAD(P)H oxidase, and tiron, superoxide scavenger, inhibited Ang II- and Ang II+ IL-1beta-stimulated increases in OPN mRNA. Ang II activated ERK1/2 within 5 min of treatment, not JNKs. IL-1beta activated ERK1/2 and JNKs within 15 min of treatment. A combination of Ang II and IL-1beta activated ERK1/2 within 5 min of treatment. None of these stimuli activated p38 kinase. DPI almost completely inhibited Ang II + IL-1beta-stimulated activation of ERK1/2, while partially inhibiting JNKs. PD98059, ERK1/2 pathway inhibitor, and SP600125, JNKs inhibitor, partially inhibited Ang II + IL-1beta-stimulated increases in OPN mRNA. A combination of PD98059 and SP600125 almost completely inhibited Ang II + IL-1beta-stimulated increases in OPN mRNA. Thus, Ang II alone increases OPN expression, while IL-1beta and TNF-alpha act synergistically with Ang II to increase OPN mRNA possibly via NO independent mechanisms. The synergistic increase in OPN mRNA involves ROS-mediated activation of ERK1/2 and JNKs, not P38 kinase, pathways in cardiac fibroblasts. PMID- 14755546 TI - Differentiation of H9c2 cardiomyoblasts: The role of adenylate cyclase system. AB - The adenylate cyclase (AC)/cAMP/cAMP-dependent protein kinase pathway controls many biological phenomena. The molecular mechanisms by which cAMP induces alternative commitment towards differentiation or proliferation are not still completely known. The differentiation of myoblast cell lines into myocytes/myotubes represents a well-established model of skeletal muscle differentiation. We analyzed the AC/cAMP pathway during terminal differentiation of H9c2 myoblasts. When cultured in low-serum containing medium, H9c2 myoblasts exit the cell cycle and differentiate into myocytes/myotubes. A key step of this process is the expression of myogenin, an essential transcription factor for the terminal differentiation into myocytes. During this phenomenon we observed a decrease in both cAMP levels and AC activity, which suggests a functional negative role of cAMP on the differentiation process of H9c2 cells. 8-Br-cAMP and other cAMP-elevating agents, such as forskolin, IBMX, and isoproterenol, negatively affected skeletal muscle differentiation of H9c2 myoblasts. Both AC activity down-regulation and intracellular cAMP reduction were accompanied by significant variations in the levels of membrane proteins belonging to the AC system (AC catalytic subunit, G(alphai-1), G(alphas)). The functional relationship between intracellular cAMP content and protein levels of AC system is discussed. PMID- 14755547 TI - ERK1/2 mediates TNF-alpha-induced matrix metalloproteinase-9 expression in human vascular smooth muscle cells via the regulation of NF-kappaB and AP-1: Involvement of the ras dependent pathway. AB - The expression of matrix metalloproteinase-9 (MMP-9) has been implicated in progression of atherosclerotic lesions. The role and importance of the signaling pathway in the transcriptional regulation of MMP-9 in human aortic smooth muscle cells (HASMC) was examined. Tumor necrosis factor-alpha (TNF-alpha) stimulated the secretion of MMP-9 in HASMC, as shown by zymography and immunoblot analysis. At the transcriptional levels, TNF-alpha also stimulated the 5'-flanking 710-bp promoter activity of MMP-9. Transcription factors NF-kappaB binding site (-601) and AP-1 binding site (-82) were identified as the cis-elements for TNF-alpha activation, as determined by gel shift assay and mutation analysis. Treatment with U0126, an inhibitor of the extracellular signal-regulated kinase (ERK), significantly downregulated TNF-alpha-induced MMP-9 expression and promoter activity, whereas the inactive analog U0124 had no effect. Furthermore, the transactivation of TNF-alpha-stimulated NF-kappaB and AP-1 was inhibited by U0126 treatment. Finally, the transient transfection of HASMC with dominant negative Ras (RasN17) suppressed TNF-alpha-induced ERK activity, MMP-9 production, and promoter activity. Overexpression of RasN17 also abolished the TNF-alpha stimulated NF-kappaB and AP-1 activity. In conclusion, the findings herein indicate the activation of the Ras/ERK pathway contributes to the induction of MMP-9 expression in HASMC. In addition, the transcription factors NF-kappaB and AP-1 that are involved in the Ras/ERK-mediated control of MMP-9 regulation on HASMC in response to TNF-alpha have now been identified. PMID- 14755548 TI - CREB Cooperates with BMP-stimulated Smad signaling to enhance transcription of the Smad6 promoter. AB - Growth plate chondrocytes integrate a multitude of growth factor signals during maturation. PTHrP inhibits maturation through stimulation of PKA/CREB signaling while the bone morphogenetic proteins (BMPs) stimulate maturation through Smad mediated signaling. In this manuscript, we show that interactions between CREB and the BMP associated Smads are promoter specific, and demonstrate for the first time the requirement of CREB signaling for Smad mediated activation of a BMP responsive region of the Smad6 promoter. The 28 base pairs (bp) BMP responsive element of the Smad6 promoter contains an 11 bp Smad binding region and an adjacent 17 bp region in which we characterize a putative CRE site. PKA/CREB gain of function enhanced BMP stimulation of this reporter, while loss of CREB function diminished transcriptional activity. In contrast, ATF-2 and AP-1 transcription factors had minimal effects. Electrophoretic mobility shift assay (EMSA) confirmed CREB binding to the Smad6 promoter element. Mutations eliminating binding resulted in loss of transcriptional activity, while mutations that maintained CREB binding had continued reporter activation by CREB and BMP-2. The Smad6 gene was similarly regulated by CREB. Dominant negative CREB reduced BMP-2 stimulated Smad6 gene transcription by 50%, but markedly increased BMP-2 mediated stimulation of colX and Ihh expression. In contrast, PTHrP which activates CREB signaling, blocked the stimulatory effect of BMP-2 on colX and Ihh, but minimally inhibited the stimulatory effect of BMP on Smad6. These findings are the first to demonstrate a cooperative association between CREB and BMP regulated Smads in cells from vertebrates and demonstrate that promoter specific rather than generalized interactions between PKA/CREB and BMP signaling regulate gene expression in chondrocytes. PMID- 14755549 TI - Enterocytin: A new specific enterocyte marker bearing a B30.2-like domain. AB - Enterocyte differentiation is correlated to the expression of specific proteins which only a few of them are identified. In this study, we characterize a new marker of enterocyte differentiation using monoclonal antibodies. We showed that small intestinal enterocytes specifically express a new 47 kDa protein named Enterocytin. Expression of this protein increase along the crypt-villus axis and it is concentrated in the terminal web, lateral plasma membrane domain, and nucleus membrane of mature enterocytes. A 1.8-kb cDNA of Enterocytin was isolated by expression cloning from a cDNA library of rabbit small intestine. The amino acid sequence obtained shows an N-terminal region with a coiled-coil structure and a B30.2-like domain in the C-terminus region. By co-transfection and immunoprecipitation procedures on Cos cells, it was observed that the coiled-coil domain is involved in the homodimerization of Enterocytin. In the human intestine, a similar 47 kDa protein was detected, exclusively in the small intestinal enterocytes. In addition, expression of this protein in Caco2 cells is correlated with the state of differentiation of these cells. The restricted expression of Enterocytin in the intestine and its localization in mature cells suggest that it may contribute to the differentiation processes and maintenance of the enterocytic polarity. PMID- 14755550 TI - Induction of apoptosis by a hepatocyte conditioned medium. AB - Incubation of primary cultures of parenchymal hepatocytes in a conditioned medium (CM), collected over the first 3 h of serum-free rat hepatocyte culture (CM(0 3)), induces a time dependent increase of the frequency of apoptotic cells which is accompanied by prominent changes of cell morphology. Short-term treatment with CM(0-3) for the first 3 h of culture is sufficient to significantly (P < 0.05) increase the frequency of apoptotic cells, however, the effect is more pronounced upon long-term treatment. Although apoptosis induction by CM(0-3) is independent of the timepoint when cultivation in CM(0-3) starts, our results suggest that the sensitivity for apoptosis induction by CM(0-3) is increased during the phase of attachment. Purification of CM(0-3) resulted in a fraction which significantly (P < 0.05) induced apoptosis at concentrations >/=10 ng/ml. Exposure of cultures to concentrations >/=1 microg/ml of purified CM(0-3) gave rise to a prominent cytotoxic effect as indicated by the massive occurrence of necrotic cells. Biochemical analysis showed that the purified fraction of CM(0-3) contains acidic ferritins with molecular weight of 23 and 43 kDa. Strikingly, both share homologies with placental isoferritins (PLF), for which growth inhibitory and immunosuppressive effects have been demonstrated by several investigations. Therefore, our results provide evidence that rat hepatocytes produce PLF or PLF related acidic isoferritins which are able to induce apoptosis. PMID- 14755551 TI - Caldesmon phosphorylation is catalyzed by two kinases in permeabilized and intact vascular smooth muscle. AB - Smooth muscle contraction is initiated by myosin light chain (MLC) phosphorylation catalyzed by the Ca(2+) dependent MLC kinase. However, many aspects of smooth muscle contraction cannot be accounted for by MLC phosphorylation. One hypothesis that has received experimental support involves the thin filament protein caldesmon. Caldesmon inhibits myosin ATPase activity; phosphorylation of caldesmon relieves this inhibitory effect. The primary candidates for catalysis of caldesmon phosphorylation are the p42/p44 ERK MAP kinases. However, we and others have shown that inhibition of the ERK MAP kinases has no effect on many smooth muscles. The goal of this study was to determine if evidence for a second endogenous caldesmon kinase may be obtained. We used Triton X-100 skinned and intact tissues of the swine carotid artery to address this goal. Caldesmon phosphorylation was evident in resting and Ca(2+) stimulated Triton X-100 skinned fibers. Ca(2+)-dependent caldesmon phosphorylation was partially sensitive to the ERK MAP kinase inhibitor PD98059, whereas all caldesmon phosphorylation was sensitive to the general kinase inhibitor, staurosporine. Histamine increased caldesmon phosphorylation levels in intact swine carotid artery, which was sensitive to both PD98059 and staurosporine. Histamine increased ERK MAP kinase activity, which was reversed by PD98059, staurosporine, and EGTA. Histamine-induced contractions were inhibited by staurosporine but not by PD98059. We interpret these results to suggest that although ERK MAP kinases catalyze caldesmon phosphorylation, a second staurosporine sensitive kinase is also important in caldesmon phosphorylation and it is this pathway that may be more important in contractile regulation. PMID- 14755552 TI - PDGF-BB enhances alpha1beta1 integrin-mediated activation of the ERK/AP-1 pathway involved in collagen matrix remodeling by rat mesangial cells. AB - Platelet-derived growth factor-BB (PDGF-BB) has been implicated in the pathogenesis of progressive glomerulonephritis (GN). Previous studies have reported that PDGF-BB stimulates mesangial cells (MCs)-induced collagen matrix remodeling through enhancement of alpha1beta1 integrin-dependent migratory activity. To determine the cell signaling pathway responsible for abnormal MC related mesangial matrix remodeling in progressive GN, we studied the involvement of the extracellular signal-regulated kinase (ERK)/activator protein-1 (AP-1) pathway in PDGF-BB-enhanced collagen gel contraction. Western blotting and gel shift assay revealed that MC-induced gel contraction resulted in ERK activation in parallel with that of AP-1 binding, peaking at 4 h and lasting at least for 24 h. Application of the MEK inhibitor, U0126, and the c-jun/AP-1 inhibitor, curcumin, inhibited gel contraction and AP-1 activity, respectively, dose dependently. PDGF-BB enhanced not only gel contraction but ERK phosphorylation and AP-1 activity by MCs. Marked inhibitory effects on PDGF-BB-induced gel contraction and ERK/AP-1 activity were observed in the presence of either function blocking anti-alpha1- or anti-beta1-integrin antibody or U0126. Consistently, AP-1-inactive MCs expressing a dominant-negative mutant of c-jun showed a significant decrease of PDGF-BB-induced gel contraction as compared with mock-transfected MCs. Finally, migration assay showed that ERK/AP-1 activity is required for PDGF-BB-stimulated alpha1beta1 integrin-dependent MC migration to collagen I. These results indicated that PDGF-BB enhances alpha1beta1 integrin mediated collagen matrix reorganization through the activation of the ERK/AP-1 pathway that is crucial for MC migration. We conclude that the ERK/AP-1 pathway plays an important role in PDGF-BB-induced alpha1beta1 integrin-dependent collagen matrix remodeling; therefore, the inhibition of its pathway may provide a novel approach to regulate abnormal collagen matrix remodeling in progressive GN. PMID- 14755554 TI - Effects of gene dosage, promoters, and substrates on unfolded protein stress of recombinant Pichia pastoris. AB - The expression of heterologous proteins may exert severe stress on the host cells at different levels. Depending on the specific features of the product, different steps may be rate-limiting. For the secretion of recombinant proteins from yeast cells, folding and disulfide bond formation were identified as rate-limiting in several cases and the induction of the chaperone BiP (binding protein) is described. During the development of Pichia pastoris strains secreting human trypsinogen, a severe limitation of the amount of secreted product was identified. Strains using either the AOX1 or the GAP promoter were compared at different gene copy numbers. With the constitutive GAP promoter, no effect on the expression level was observed, whereas with the inducible AOX1 promoter an increase of the copy number above two resulted in a decrease of expression. To identify whether part of the product remained in the cells, lysates were fractionated and significant amounts of the product were identified in the insoluble fraction containing the endoplasmic reticulum, while the soluble cytosolic fraction contained product only in clones using the GAP promoter. An increase of BiP was observed upon induction of expression, indicating that the intracellular product fraction exerts an unfolded protein response in the host cells. A strain using the GAP promoter was grown both on glucose and methanol and trypsinogen was identified in the insoluble fractions of both cultures, but only in the soluble fraction of the glucose grown cultures, indicating that the amounts and distribution of intracellularly retained product depends on the culture conditions, especially the carbon source. PMID- 14755555 TI - Microbioreactor arrays with parametric control for high-throughput experimentation. AB - A scalable array technology for parametric control of high-throughput cell cultivations is demonstrated. The technology makes use of commercial printed circuit board (PCB) technology, integrated circuit sensors, and an electrochemical gas generation system. We present results for an array of eight 250 microl microbioreactors. Each bioreactor contains an independently addressable suite that provides closed-loop temperature control, generates feed gas electrochemically, and continuously monitors optical density. The PCB technology allows for the assembly of additional off-the-shelf components into the microbioreactor array; we demonstrate the use of a commercial ISFET chip to continuously monitor culture pH. The electrochemical dosing system provides a powerful paradigm for reproducible gas delivery to high-density arrays of microreactors. Growth data are presented for Escherichia coli cultured in the array with varying microaerobic conditions using electrochemically generated oxygen. Additionally, we present data on carbon dioxide generation for pH dosing. PMID- 14755556 TI - Analyzing and modeling of photobioreactors by combining first principles of physiology and hydrodynamics. AB - Mixing in photobioreactors is known to enhance biomass productivity considerably, and flow dynamics play a significant role in the reactor's performance, as they determine the mixing and the cells' movement. In this work we focus on analyzing the effects of mixing and flow dynamics on the photobioreactor performance. Based on hydrodynamic findings from the CARPT(Computer Automated Radioactive Particle Tracking) technique, a possible mechanism for the interaction between the mixing and the physiology of photosynthesis is presented, and the effects of flow dynamics on light availability and light intensity fluctuation are discussed and quantitatively characterized. Furthermore, a dynamic modeling approach is developed for photobioreactor performance evaluation, which integrates first principles of photosynthesis, hydrodynamics, and irradiance distribution within the reactor. The results demonstrate the reliability and the possible applicability of this approach to commercially interesting microalgae/cyanobacteria culture systems. PMID- 14755558 TI - Resonance energy transfer for assessing the molecular integrity of proteins for local delivery. AB - It remains unclear whether the limitations to the therapeutic potential of angiogenic growth factors stem from pharmacokinetic concerns related to inadequate delivery or from a reduced sensitivity of target tissues. Here, we report a novel method using resonance energy transfer to assess the molecular integrity of proteins after local delivery. As an example, we labeled fibroblast growth factor-2 with a fluorescent donor and nonfluorescent acceptor pair, tetramethylrhodamine and QSY-7, and demonstrate in an ex vivo bovine carotid artery model that this growth factor is not limited by proteolytic constraints imposed by the tissue. Our data indicate that FGF-2 is unlikely to be degraded within the arterial wall and suggest that pharmacokinetic limitations alone cannot fully explain the muted response seen thus far in therapeutic angiogenesis. In general, resonance energy transfer may serve as a novel approach to assess the molecular integrity of protein-based therapies in local delivery. PMID- 14755557 TI - Characterization of the cellular uptake and metabolic conversion of acetylated N acetylmannosamine (ManNAc) analogues to sialic acids. AB - "Sialic acid engineering" refers to the strategy where cell surface carbohydrates are modified by the biosynthetic incorporation of metabolic intermediates, such as non-natural N-acetylmannosamine (ManNAc) analogues, into cellular glycoconjugates. While this technology has promising research, biomedical, and biotechnological applications due to its ability to endow the cell surface with novel physical and chemical properties, its adoption on a large scale is hindered by the inefficient metabolic utilization of ManNAc analogues. We address this limitation by proposing the use of acetylated ManNAc analogues for sialic acid engineering applications. In this paper, the metabolic flux of these "second generation" compounds into a cell, and, subsequently, into the target sialic acid biosynthetic pathway is characterized in detail. We show that acetylated ManNAc analogues are metabolized up to 900-fold more efficiently than their natural counterparts. The acetylated compounds, however, decrease cell viability under certain culture conditions. To determine if these toxic side effects can be avoided, we developed an assay to measure the cellular uptake of acetylated ManNAc from the culture medium and its subsequent flux into sialic acid biosynthetic pathway. This assay shows that the majority ( > 80%) of acetylated ManNAc is stored in a cellular "reservoir" capable of safely sequestering this analogue. These results provide conditions that, from a practical perspective, enable the acetylated analogues to be used safely and efficaciously and therefore offer a general strategy to facilitate metabolic substrate-based carbohydrate engineering efforts. In addition, these results provide fundamental new insights into the metabolic processing of non-natural monosaccharides. PMID- 14755559 TI - Effect of initial moisture content and chip size on the bioconversion efficiency of softwood lignocellulosics. AB - Previous optimization strategies for the bioconversion of lignocellulosics by steam explosion technologies have focused on the effects of temperature, pH, and treatment time, but have not accounted for changes in severity brought about by properties inherent in the starting feedstock. Consequently, this study evaluated the effects of chip properties, feedstock size (40-mesh, 1.5 x 1.5 cm, 5 x 5 cm), and moisture content (12% and 30%) on the overall bioconversion process, and more specifically on the efficacy of removal of recalcitrant lignin from the lignocellulosic substrates following steam explosion. Increasing chip size resulted in an improvement in the solids recovery, with concurrent increases in the water soluble, hemicellulose-derived sugar recovery (7.5%). This increased recovery is a result of a decrease in the "relative severity" of the pretreatment as chip size increases. Additionally, the decreased relative severity minimized the condensation of the recalcitrant residual lignin and therefore increased the efficacy of peroxide fractionation, where a 60% improvement in lignin removal was possible with chips of larger initial size. Similarly, increased initial moisture content reduced the relative severity of the pretreatment, generating improved solids and hemicellulose-derived carbohydrate recovery. Both increased chip size and higher initial moisture content results in a substrate that performs better during peroxide delignification, and consequently enzymatic hydrolysis. Furthermore, a post steam-explosion refining step increased hemicellulose-derived sugar recovery and was most effectively delignified (to as low as 6.5%). The refined substrate could be enzymatically hydrolyzed to very high levels (98%) and relatively fast rates (1.23 g/L/h). PMID- 14755560 TI - Online detection of feed demand in high cell density cultures of Escherichia coli by measurement of changes in dissolved oxygen transients in complex media. AB - A starvation-based dissolved oxygen (DO) transient controller was developed to supply growth-limiting substrate to high cell density fed-batch cultures of recombinant Escherichia coli. The algorithm adjusted a preexisting feed rate in proportion to the culture's oxygen demand, which was estimated from transients in the DO concentration after short periods of feed interruption. In this manner, the addition of glucose feed was precisely controlled at a rate that did not exceed the acetate production threshold, thus preventing acetate accumulation. In comparison to exponential feed algorithms commonly used in industry, the implementation of the new feeding strategy increased the final cell density from 32 to 44 g (dry cell weight).L(-1), with less than 16 mM acetate accumulated, producing an ideal culture for subsequent induction. Despite a constant starvation level and relatively low levels of acetate, experimental cultivations still tended to produce acetate towards the end of the process. The use of a simple Monod model provided an explanation as to why this may occur in high cell density cultivations and suggests how it may be overcome. PMID- 14755561 TI - Effect of cobalt on the anaerobic thermophilic conversion of methanol. AB - The importance of cobalt on the anaerobic conversion of methanol under thermophilic conditions was studied in three parallel lab-scale UASB-reactors and in cobalt-limited enriched cultures. Reactors R1, R2, and R3 were fed with methanol in a bicarbonate-buffered medium, supplied with iron and macronutrients: in R1 all metals were supplied (control), R2 was cobalt deprived, and in R3 all metals were deprived. In the 136 days of continuous experiment, a drop in performance was observed over the last 30 days. Particularly in R3, both methanol removal and methane formation dropped by 7.1% and 13.7%, respectively, compared to the control reactor, R1. When the medium was cobalt-deprived, acetate was not produced and, as a consequence, the enriched consortium lost its capacity to degrade acetate, indicating that the acetotrophic microorganisms were washed out. The addition of 0.5 microM of cobalt to a cobalt-deprived enrichment culture led to acetate accumulation. The results obtained in this study indicate that the mixed consortium requires a proper amount of cobalt, and its addition to a concentration of 0.1 microM leads to the highest methanol conversion rate, with methane as the sole end product from methanol. PMID- 14755562 TI - Effect of immobilization support, water activity, and enzyme ionization state on cutinase activity and enantioselectivity in organic media. AB - We studied the reaction between vinyl butyrate and 2-phenyl-1-propanol in acetonitrile catalyzed by Fusarium solani pisi cutinase immobilized on zeolites NaA and NaY and on Accurel PA-6. The choice of 2-phenyl-1-propanol was based on modeling studies that suggested moderate cutinase enantioselectivity towards this substrate. With all the supports, initial rates of transesterification were higher at a water activity (a(w)) of 0.2 than at a(w) = 0.7, and the reverse was true for initial rates of hydrolysis. By providing acid-base control in the medium through the use of solid-state buffers that control the parameter pH-pNa, which we monitored using an organo-soluble chromoionophoric indicator, we were able, in some cases, to completely eliminate dissolved butyric acid. However, none of the buffers used were able to improve the rates of transesterification relative to the blanks (no added buffer) when the enzyme was immobilized at an optimum pH of 8.5. When the enzyme was immobilized at pH 5 and exhibited only marginal activity, however, even a relatively acidic buffer with a pK(a) of 4.3 was able to restore catalytic activity to about 20% of that displayed for a pH of immobilization of 8.5, at otherwise identical conditions. As a(w) was increased from 0.2 to 0.7, rates of transesterification first increased slightly and then decreased. Rates of hydrolysis showed a steady increase in that a(w) range, and so did total initial reaction rates. The presence or absence of the buffers did not impact on the competition between transesterification and hydrolysis, regardless of whether the butyric acid formed remained as such in the reaction medium or was eliminated from the microenvironment of the enzyme through conversion into an insoluble salt. Cutinase enantioselectivity towards 2-phenyl-1 propanol was indeed low and was not affected by differences in immobilization support, enzyme protonation state, or a(w). PMID- 14755563 TI - Biocatalytic scrubbing of gaseous acrylonitrile using Rhodococcus ruber immobilized in synthetic silicone polymer (ImmobaSil) rings. AB - Microbial detoxification of acrylonitrile vapor was performed using Rhodococcus ruber immobilized into rings of a synthetic silicone polymer (ImmobaSil ) sponge. The immobilization matrix was used as both the cell support material and also as the trickle-bed column packing. The elimination capacity of the biocatalytic scrubber was determined for different influent concentrations of gaseous acrylonitrile. The working life of the nongrowing biocatalyst was also examined under test conditions and was found to be at least 70 days before biocatalyst replacement was required. An elimination capacity of 4.0 kg m(-3) h(-1) was achieved with a 95% removal efficiency, and an elimination capacity of over 7.2 kg m(-3) h(-1) acrylonitrile was achieved with a removal efficiency of 90%. This elimination capacity is 10 times that previously published in the literature. PMID- 14755564 TI - Salt-activation of nonhydrolase enzymes for use in organic solvents. AB - Enzymatic reactions are important for the synthesis of chiral molecules. One factor limiting synthetic applications of enzymes is the poor aqueous solubility of numerous substrates. To overcome this limitation, enzymes can be used directly in organic solvents; however, in nonaqueous media enzymes usually exhibit only a fraction of their aqueous-level activity. Salt-activation, a technique previously demonstrated to substantially increase the transesterification activity of hydrolytic enzymes in organic solvents, was applied to horse liver alcohol dehydrogenase, soybean peroxidase, galactose oxidase, and xanthine oxidase, which are oxidoreductase and oxygenase enzymes. Assays of the lyophilized enzyme preparations demonstrated that the presence of salt protected enzymes from irreversible inactivation. In organic solvents, there were significant increases in activity for the salt-activated enzymes compared to nonsalt-activated controls for every enzyme tested. The increased enzymatic activity in organic solvents was shown to result from a combination of protection against inactivation during the freeze-drying process and other as-yet undetermined factors. PMID- 14755565 TI - Excluded volume effect in unzipping DNA with a force. AB - A double stranded DNA molecule when pulled with a force acting on one end of the molecule can become either partially or completely unzipped depending on the magnitude of the force F. For a random DNA sequence, the number M of unzipped base pairs goes as M approximately (F - Fc)(-2) and diverges at the critical force Fc with an exponent chi = 2. We find that when excluded volume effect is taken into account for the unzipped part of the DNA, the exponent chi = 2 is not changed but the critical force Fc is changed. The force versus temperature phase diagram depends on only two parameters in the model, the persistence length and the denaturation temperature. Furthermore a scaling form of the phase diagram can be found. This scaling form is parameter independent and depends only on the spatial dimension. It applies to all DNA molecules and should provide a useful framework for comparison with experiments. PMID- 14755566 TI - Molecular dynamics simulations of a guaiacyl beta-O-4 lignin model compound: examination of intramolecular hydrogen bonding and conformational flexibility. AB - The dynamical conformational behavior of a guaiacyl beta-O-4 lignin model compound has been investigated by molecular simulations. The potential energy surface of the molecule in vacuum has been examined by means of an adiabatic map, showing a large accessible conformational space with multiple energy minima separated by low barriers. Molecular dynamics simulations have been performed in vacuum and with explicit solvent molecules for 10 and 2.1 ns, respectively. Molecular dynamics trajectories recorded in vacuum have shown the molecule to be flexible and to visit a large number of conformations. Many intramolecular H bonds have been observed, existing for more than 90% of the total simulation time. The presence of explicit solvent molecules induces a significant broadening of some regions of the accessible conformational space and also largely reduces the statistical significance of intramolecular H-bonding. Intramolecular H-bonds observed in vacuum do not persist significantly and are preferentially exchanged with intermolecular H-bonds to the surrounding solvent molecules. The theoretical results are in good agreement with experimental NMR data that do not support the existence of strong and persistent intramolecular H-bonds in solution but instead indicate that H-bonds to solvent predominate. Finally, both molecular modeling and NMR approaches predict the guaiacyl beta-O-4 structure to be flexible and indicate that intramolecular H-bonds are not strong and persistent enough to confer rigidity to the molecule in solution. PMID- 14755567 TI - Structural similarity of E. coli 5S rRNA in solution and within the ribosome. AB - The article presents translational and rotational diffusion coefficients of 5S rRNA determined experimentally by the method of dynamic light scattering (DLS) and its comparison with the values predicted for different models of this molecule. The tertiary structure of free 5S rRNA was proposed on the basis of the atomic structures of the 5S rRNA from E. coli and H. marismortui extracted from the ribosome. A comparison of the values of DT, tauR, and Rg predicted for different models with experimental results for the free molecule in solution suggests that free 5S rRNA is less compact than that in the complex with ribosomal proteins. In general, the molecules of 5S rRNA consist of three domains: a short one and two longer ones. As follows from a comparison of the results of our simulations with experimental values, in the molecule in solution the two closest helical fragments of the longer domains remain collinear, whereas the short domain takes a position significantly deviated from them. PMID- 14755569 TI - DNA sequence-dependent deformability--insights from computer simulations. AB - The article reviews some recent developments in studying DNA sequence-dependent deformability, with emphasis on computer modeling. After a brief outline of available experimental techniques, we proceed to computational methods and focus on atomic-resolution molecular dynamics (MD) simulations. A sequence-dependent local (base-pair step) force field inferred from MD is compared with force fields obtained by other techniques. Various methods for establishing global (flexible rod) DNA elastic constants are reviewed, including an approach based on atomic resolution MD. The problem of defining the global deformation variables, as well as the question of anisotropy and nonlocal effects, are discussed. As an example, both local and global deformability calculations from atomic-resolution MD of EcoRI dodecamer are presented. PMID- 14755570 TI - Structural motifs in ribosomal RNAs: implications for RNA design and genomics. AB - The various motifs of RNA molecules are closely related to their structural and functional properties. To better understand the nature and distributions of such structural motifs (i.e., paired and unpaired bases in stems, junctions, hairpin loops, bulges, and internal loops) and uncover characteristic features, we analyze the large 16S and 23S ribosomal RNAs of Escherichia coli. We find that the paired and unpaired bases in structural motifs have characteristic distribution shapes and ranges; for example, the frequency distribution of paired bases in stems declines linearly with the number of bases, whereas that for unpaired bases in junctions has a pronounced peak. Significantly, our survey reveals that the ratio of total (over the entire molecule) unpaired to paired bases (0.75) and the fraction of bases in stems (0.6), junctions (0.16), hairpin loops (0.12), and bulges/internal loops (0.12) are shared by 16S and 23S ribosomal RNAs, suggesting that natural RNAs may maintain certain proportions of bases in various motifs to ensure structural integrity. These findings may help in the design of novel RNAs and in the search (via constraints) for RNA-coding motifs in genomes, problems of intense current focus. PMID- 14755571 TI - Packaging double-helical DNA into viral capsids. AB - DNA packaging in bacteriophage P4 has been examined using a molecular mechanics model with a reduced representation containing one pseudoatom per turn of the double helix. The model is a discretized version of an elastic continuum model. The DNA is inserted piecewise into the model capsid, with the structure being reoptimized after each piece is inserted. Various optimization protocols were investigated, and it was found that molecular dynamics at a very low temperature (0.3 K) produces the optimal packaged structure. This structure is a concentric spool, rather than the coaxial spool that has been commonly accepted for so many years. This geometry, which was originally suggested by Hall and Schellman in 1982 (Biopolymers Vol. 21, pp. 2011-2031), produces a lower overall elastic energy than coaxial spooling. PMID- 14755572 TI - DNA fine structure and dynamics in crystals and in solution: the impact of BI/BII backbone conformations. AB - Sugar-phosphate backbone conformations are an important structural element for a complete understanding of specific recognition in nucleic acid-protein interactions. They can be involved both in early stages of target discrimination and in structural adaptation upon binding. In the first part of this study, we have analyzed high-resolution structures of double-stranded B-DNA either isolated or bound to proteins, and explored the impact of both the standard BI and the unusual BII phosphate backbone conformations on neighboring sugar puckers and on selected helical parameters. Correlations are found to be similar for free and bound DNA, and in both categories, the possible facing backbone conformations (BI.BI, BI.BII, and BII.BII) define well-characterized substates in the B-DNA conformational space. Notably, BII.BII steps are characterized by specific, and sequence-independent, structural effects involving reduced standard deviations for almost all conformational parameters. In the second part of this work, we analyze four 10 ns molecular dynamics simulations in explicit solvent on the DNA targets of NF-kappaB and bovine papillomavirus E2 proteins, highlighting the multiplicity of backbone dynamical behavior. These results show sequence effects on the percentages of BI and BII conformers, the preferential state of facing backbones, the occurrence of coupled transitions. The backbone states can consequently be seen as a mechanism for transmitting information from the bases to the phosphate groups and thus for modulating the overall structural properties of the target DNA. PMID- 14755573 TI - Molecular dynamics simulations of papilloma virus E2 DNA sequences: dynamical models for oligonucleotide structures in solution. AB - The specificity of papilloma virus E2 protein-DNA binding depends critically upon the sequence of a region of the DNA not in direct contact with the protein, and represents one of the simplest known examples of indirect readout. A detailed characterization of this system in solution is important to the further investigation hypothesis of a structural code for DNA recognition by regulatory proteins. In the crystalline state, the E2 DNA oligonucleotide sequence, d(ACCGAATTCGGT), exhibits three different structural forms. We report herein studies of the structure of E2 DNA in solution based on a series of molecular dynamics (MD) simulations including counterions and water, utilizing both the canonical and various crystallographic structures as initial points of departure. All MDs converged on a single dynamical structure of d(ACCGAATTCGGT) in solution. The predicted structure is in close accord with two of the three crystal structures, and indicates that a significant kink in the double helix at the central ApT step in the other crystal molecule may be a packing effect. The dynamical fine structure was analyzed on the basis of helicoidal parameters. The calculated curvature in the sequence was found to originate primarily from YPR steps in the regions flanking the central AATT tract. In order to study the role of structural adaptation of the DNA in the binding process, a subsequent simulation on the 16-mer cognate sequence d(CAACCGAATTCGGTTG) was initiated from the crystallographic coordinates of the bound DNA in the crystal structure of the protein DNA complex. MD simulations starting with the protein-bound form relaxed rapidly back to the dynamical structure predicted from the previous simulations on the uncomplexed DNA. The MD results show that the bound form E2 DNA is a dynamically unstable structure in the absence of protein, and arises as a consequence of both structural changes intrinsic to the sequence and induced by the interaction with protein. PMID- 14755574 TI - Molecular dynamics simulations of DNA curvature and flexibility: helix phasing and premelting. AB - Recent studies of DNA axis curvature and flexibility based on molecular dynamics (MD) simulations on DNA are reviewed. The MD simulations are on DNA sequences up to 25 base pairs in length, including explicit consideration of counterions and waters in the computational model. MD studies are described for ApA steps, A tracts, for sequences of A-tracts with helix phasing. In MD modeling, ApA steps and A-tracts in aqueous solution are essentially straight, relatively rigid, and exhibit the characteristic features associated with the B'-form of DNA. The results of MD modeling of A-tract oligonucleotides are validated by close accord with corresponding crystal structure results and nuclear magnetic resonance (NMR) nuclear Overhauser effect (NOE) and residual dipolar coupling (RDC) structures of d(CGCGAATTCGCG) and d(GGCAAAAAACGG). MD simulation successfully accounts for enhanced axis curvature in a set of three sequences with phased A-tracts studied to date. The primary origin of the axis curvature in the MD model is found at those pyrimidine/purine YpR "flexible hinge points" in a high roll, open hinge conformational substate. In the MD model of axis curvature in a DNA sequence with both phased A-tracts and YpR steps, the A-tracts appear to act as positioning elements that make the helix phasing more precise, and key YpR steps in the open hinge state serve as curvature elements. Our simulations on a phased A-tract sequence as a function of temperature show that the MD simulations exhibit a premelting transition in close accord with experiment, and predict that the mechanism involves a B'-to-B transition within A-tracts coupled with the prediction of a transition in key YpR steps from the high roll, open hinge, to a low roll, closed hinge substate. Diverse experimental observations on DNA curvature phenomena are examined in light of the MD model with no serious discrepancies. The collected MD results provide independent support for the "non A-tract model" of DNA curvature. The "junction model" is indicated to be a special case of the non-A-tract model when there is a Y base at the 5' end of an A-tract. In accord with crystallography, the "ApA wedge model" is not supported by MD. PMID- 14755575 TI - Solution conformations of cyclosporins and magnesium-cyclosporin complexes determined by vibrational circular dichroism. AB - Vibrational circular dichroism (VCD) spectroscopy was used to investigate the solution conformations of cyclosporins A, C, D, G, and H in CDCl(3), in the amide I and NH/OH-stretching regions, and their corresponding magnesium complexes in CD(3)CN, in the amide I region. VCD spectra are sensitive to the chiral arrangement of Cdbond;O and NH bonds in this cyclic undecapeptide. Calculations of molecular geometries, as well as IR and VCD intensities of model cyclosporin fragments that include the intramolecular hydrogen bonds of the crystal conformations of cyclosporins A and H (CsA and CsH), were carried out at the density functional theory (DFT; BPW91 functional/6-31G* basis set) level. The good agreement between IR and VCD spectra from experiment and DFT calculations provides evidence that the crystal conformation of CsA is dominant in CDCl(3) solution; CsH, however, assumes both an intramolecularly hydrogen-bonded crystal conformation and more open forms in solution. Comparisons of the experimental and calculated VCD spectra in the NH/OH-stretching region of the noncomplexed cyclosporins indicate that conformers with both free and hydrogen-bonded NH and OH groups are present in solution. Differences between the IR and VCD spectra for the metal-free and magnesium-complexed cyclosporins are indicative of strong interactions between cyclosporins and magnesium ions. PMID- 14755576 TI - The ribbon of hydrogen bonds in globular proteins. IV. The example of the papain family. AB - A study of the role of the hydrogen-bonding side chains in the ribbon of hydrogen bonds in globular proteins, using the papain family as an example, suggests that these side chains may be divided into three categories depending on their position in the molecule. In the first category, they form part of the local ribbon, in the second they form part of the ribbon at a site remote along the main chain, and in the third they play no role in the formation of the ribbon. The second case is particularly interesting because it provides a natural mechanism for the formation of the tertiary structure of the globular proteins. The results suggest that the robustness of the globular proteins towards mutations arises from the fact that many mutations that involve hydrogen-bonding side chains either leave the hydrogen bonding of the ribbon essentially unchanged or their hydrogen bonding plays no part in the formation of the ribbon in the first place. The results show that it is possible to obtain the ribbon of hydrogen bonds for a family of proteins whose data set's are of intermediate quality by studying the ribbons of several members of such a family and then taking an average over the different partial ribbons to create a standard ribbon of hydrogen bonds for the family as a whole. This method is used here to derive the standard ribbon for the papain family with papain itself, actinidin, and human liver cathepsin B as the representatives of the family. All three members of the family fit the standard ribbon with an accuracy of 85-91%. This result opens up the use of this technique for the study of a large number of globular proteins whose recorded data sets are of intermediate quality. PMID- 14755577 TI - Calculation of ligand-nucleic acid binding free energies with the generalized born model in DOCK. AB - The calculation of ligand-nucleic acid binding free energies is investigated by including solvation effects computed with the generalized-Born model. Modifications of the solvation module in DOCK, including introduction of all-atom parameters and revision of coefficients in front of different terms, are shown to improve calculations involving nucleic acids. This computing scheme is capable of calculating binding energies, with reasonable accuracy, for a wide variety of DNA ligand complexes, RNA-ligand complexes, and even for the formation of double stranded DNA. This implementation of GB/SA is also shown to be capable of discriminating strong ligands from poor ligands for a series of RNA aptamers without sacrificing the high efficiency of the previous implementation. These results validate this approach to screening large databases against nucleic acid targets. PMID- 14755578 TI - Identification of possible kinetically significant anion-binding sites in human serum transferrin using molecular modeling strategies. AB - Certain anions have been shown experimentally to influence the rate of iron release from human serum transferrin (HST), implying the existence of one or more allosteric kinetically significant anion-binding (KISAB) sites on or near the surface of the protein. A rank-ordered selection of potential HST KISAB sites has been obtained using a novel three-stage molecular modeling strategy. The crystal structure of HST (1A8E.pdb) was first subjected to a heuristic analysis, in which positively charged and hydrogen-bonding residues on or near the surface of the protein were identified. In this stage, a preliminary electrostatic potential map was also calculated, yielding six preliminary sites. Next, energy-grid calculations were conducted in order to identify anion-protein interaction energy minima, which resulted in the inclusion of three additional sites. Finally, three anions already shown experimentally to demonstrate varied effects on HST iron release kinetics were placed at each potential site; molecular dynamics and molecular mechanics calculations were performed in order to elucidate the hydrogen-bonding environment around each anion of the protein as well as to calculate anion-protein-binding energies. PMID- 14755579 TI - DNA melting profiles from a matrix method. AB - In this article we give a new method for the calculation of DNA melting profiles. Based on the matrix formulation of the DNA partition function, the method relies for its efficiency on the fact that the required matrices are very sparse, essentially reducing matrix multiplication to vector multiplication and thus making the computer time required to treat a DNA molecule containing N base pairs proportional to N(2). A key ingredient in the method is the result that multiplication by the inverse matrix can also be reduced to vector multiplication. The task of calculating the melting profile for the entire genome is further reduced by treating regions of the molecule between helix-plateaus, thus breaking the molecule up into independent parts that can each be treated individually. The method is easily modified to incorporate changes in the assignment of statistical weights to the different structural features of DNA. We illustrate the method using the genome of Haemophilus influenzae. PMID- 14755580 TI - Conformational and structural analysis of the equilibrium between single- and double-strand beta-helix of a D,L-alternating oligonorleucine. AB - Alternating sequences of D and L residues in peptides are directly related to the formation of several kinds of regular helical conformations usually called beta helices. The major feature of these structures is that they can be associated with the transmembrane ion-conducting channel activity in some natural antibacterial peptides. The study of alternating D,L synthetic peptides is critical to understand how factors such as surrounding media, main chain length, type of side chain and terminal groups, among others, can determine the adoption of a specific kind of beta-helix. Early studies pointed out that the peptides Boc (D-NLeu-L-NLeu)(6)-D-MeNLe-L-Nl-D-Nl-L-Nl-OMe (Boc: tert-butyloxycarbonyl) and Boc-L-Nle-(D-Nle-L-Nle)(5)-D-MeNle-L-Nle-D-Nle-L-Nle-OMe adopt in chloroform a unique detectable conformation single beta(4.4)- and double beta(5.6) upward arrow downward arrow -helix, respectively. The influence of terminal groups on the final stable conformation of N-formylated peptides has been studied in this work. The initial basic NMR data analysis of a synthetic alternating D,L oligopeptide with ten norleucines, N-methylated on the residue 7 and having HCO- and -OMe as terminal groups clearly indicates the coexistence of two different conformations in equilibrium. NMR data and molecular dynamics calculations point to a dimeric antiparallel beta-helix structure beta(5.6) upward arrow downward arrow for the main conformation. On the other hand, NMR data suggest a single beta-helix structure beta(4.4) for the second conformation. Finally, a thermodynamic analysis of the equilibrium between both conformations has been carried out by one-dimensional NMR measurements at ten different temperatures. The temperature at which 50% of dimer conformation is dissociated is 319 K. In addition, the dimer-monomer equilibrium curve obtained shows a DeltaG>0 for the whole range of studied temperatures, and its behavior can be considered similar to the thermodynamic denaturation protein processes. PMID- 14755581 TI - Volumetric properties of the formation of double stranded DNA: a nearest-neighbor analysis. AB - The kinetics of the helix-coil transition have been studied by performing UV monitored melting and reannealing curves of DNA and analyzing the resultant hysteresis between these curves. The analysis assumes a single-step bimolecular transition with duplex formation defined as the forward reaction. Volume parameters of the helix-coil transition were obtained by measuring the pressure dependence of the rate constants from 5-200 MPa. The data were interpreted in terms of several possible nearest-neighbor models, ranging from one to eleven parameters. Twenty-four oligonucleotide duplexes 22 base pairs in length were used to solve for individual nearest-neighbor activation volumes and transition volumes. Statistically, the most valid fit of the volumetric data was obtained with a six-parameter model in which the directionality of the dinucleotide steps is not considered, for example, 5'AG/CT is the same as 5'GA/TC. The resultant transition volumes at 48 degrees C ranged from -7.1 +/- 0.8 mL/mol (GC/CG) to +2.9 +/- 0.3 mL/mol (AA/TT). The success of the six-parameter model suggests that the relative size of the nearest-neighbor dinucleotides is the most important factor determining the magnitude of the volumetric parameters. The finding that the magnitude of the volumetric parameters correlates with the change in the solvent accessible surface area of the bases during the helix-coil transition corroborates this hypothesis. PMID- 14755582 TI - Exploring the interaction of the surfactant N-terminal domain of gamma-Zein with soybean phosphatidylcholine liposomes. AB - Zeins are maize storage proteins that accumulate inside large vesicles called protein bodies. gamma-Zein lines the inner surface of the protein body membrane, and its N-terminal, proline-rich, repetitive domain with the sequence (VHLPPP)(8) appears to be necessary for the accumulation of the protein within the organelle. Synthetic (VHLPPP)(8) adopts an amphipathic polyproline II conformation and forms cylindrical micelles in aqueous solution. Here we explore the interaction of (VHLPPP)(8) with soybean phosphatidylcholine unilamellar lipid vesicles and examine its effect on the stability and permeability of the liposome membrane. The amphipathic N-terminal domain of gamma-zein interacts with the membrane and assembles to form extended domains over the phospholipid membrane. The interaction between the peptide and the membrane increases the stability and permeability of the liposome membrane. The spontaneous amphipathic aggregation of (VHLPPP)(8) on the membrane suggests a mechanism of gamma-zein deposition inside maize protein bodies. PMID- 14755583 TI - Effect of phosphorothioate chirality on the grooves of DNA double helices: a molecular dynamics study. AB - Phosphorothioate oligonucleotides (PS-ODNs) have gained considerable attention in drug therapy, primarily as potent antisense or antigene oligomers, which bind to specific DNA or mRNA sequences and lead to transcriptional or translational arrest. These are obtained by substituting one of the anionic oxygen of the phosphate group by a sulfur atom, which introduces chirality to the phosphorus atom of the DNA backbone. In this molecular dynamics simulation study, structural parameters like groove widths, environmental parameters like hydration or cation binding, and electrostatic energy surfaces of both the chiral forms of DNA/PS-DNA duplexes were assessed and compared with that of a normal DNA. Results indicate that, PS-S form with its sulfur atoms facing the minor groove has a widened minor groove, while the scenario is reverse for the PS-R form. Further analysis reveals the existence of several factors like large van der Waals radius of sulfur and the effect it has on its neighboring hydration pattern along with the net electrostatic environment, influencing such structural alterations. This also indicates, for the first time, the effect of absolute phosphorothioate chirality on the global structure of a DNA/PS-DNA hybrid that otherwise resembles a regular B-DNA structure. PMID- 14755584 TI - Temperature and pressure effects on conformational equilibria of alanine dipeptide in aqueous solution. AB - We investigated the temperature and pressure effects on conformational equilibria of N-acetyl-L-alanine-N'-methylamide (AAlaMA) in aqueous solution by Raman spectroscopy. Scattering intensities in the skeletal stretching mode of AAlaMA in aqueous solution were decomposed into some component bands by the spectra analysis. Our results indicate that each component band for AAlaMA adopts not only the P(II) and alpha(R) conformations but also the C(7eq) conformation. From temperature and pressure dependencies of the band intensities, we determined the enthalpy differences and the volume differences between the conformers. The C(7eq) conformer is enthalpically most stable due to the intramolecular hydrogen bond. The partial molar volume of the C(7eq) conformer is the smallest through the solvent-exclusion effect rather than the solute-solvent electrostatic interaction effect. PMID- 14755586 TI - Abstracts of the 73rd Annual Meeting of the American Association of Physical Anthropologists. Tampa, Florida, USA, April 14-17, 2004. PMID- 14755593 TI - Changes in effective connectivity models in the presence of task-correlated motion: an fMRI study. AB - We investigated the effects of motion-correction strategy and time course selection method when structural equation modeling is applied to fMRI data in the presence of task-correlated motion. Three motion-correction methods were employed for a group of 12 subjects performing an orthographic lexical retrieval task: (1) a rigid body realignment as implemented in SPM99, (2) a rigid body realignment combined with the inclusion of motion parameters in the statistical model, and (3) the FLIRT motion correction followed by an ICA analysis aiming to identify and remove the motion-related components and the ghosting artifacts. For each motion correction, the time courses of the activated regions were selected in three ways: (1) using the voxels with the highest Z scores, (2) using the average across all the statistically significant voxels in the region of interest, and (3) using a within-region, across-subjects, singular value decomposition. The resulting models of effective connectivity were markedly different, although the activation pattern was not substantially altered by the motion-correction method. Higher values for the path coefficients were obtained for the models fitted to the covariance matrices based on the average time courses than for the covariance matrices based on a single voxel time course. Our results suggest caution with the interpretation of task-induced changes in effective connectivity since, for higher-order cognitive brain functions, multiple models can be fitted to a given data set and these models cannot be rejected on an anatomical or cognitive basis. Hum. Brain Mapping 21:49-63, 2004. PMID- 14755594 TI - Mapping the time course of nonconscious and conscious perception of fear: an integration of central and peripheral measures. AB - Neuroimaging studies using backward masking suggest that conscious and nonconscious responses to complex signals of fear (facial expressions) occur via parallel cortical and subcortical circuits. Little is known, however, about the temporal differentiation of these responses. Psychophysics procedures were first used to determine objective thresholds for both nonconscious detection (face vs. blank screen) and discrimination (fear vs. neutral face) in a backward masking paradigm. Event-related potentials (ERPs) were then recorded (n = 20) using these thresholds. Ten blocks of masked fear and neutral faces were presented under each threshold condition. Simultaneously recorded skin conductance responses (SCRs) provided an independent index of stimulus perception. It was found that Fear stimuli evoked faster SCR rise times than did neutral stimuli across all conditions, indicating that emotional content influenced responses, regardless of awareness. In the first 400 msec of processing, ERPs dissociated the time course of conscious (enhanced N4 component) from nonconscious (enhanced N2 component) perception of fear, relative to neutral. Nonconscious detection of fear also elicited relatively faster P1 responses within 100 msec post-stimulus. The N2 may provide a temporal correlate of the initial sensory processing of salient facial configurations, which is enhanced when top-down cortical feedback is precluded. By contrast, the N4 may index the conscious integration of emotion stimuli in working memory, subserved by greater cortical engagement. Hum. Brain Mapping 21:64-74, 2004. PMID- 14755595 TI - Functional brain mapping during free viewing of natural scenes. AB - Previous imaging studies have used mostly perceptually abstracted, idealized, or static stimuli to show segregation of function in the cerebral cortex. We wanted to learn whether functional segregation is maintained during more natural, complex, and dynamic conditions when many features have to be processed simultaneously, and identify regions whose activity correlates with the perception of specific features. To achieve this, we used functional magnetic resonance imaging (fMRI) to measure brain activity when human observers viewed freely dynamic natural scenes (a James Bond movie). The intensity with which they perceived different features (color, faces, language, and human bodies) was assessed psychometrically in separate sessions. In all subjects different features were perceived with a high degree of independence over time. We found that the perception of each feature correlated with activity in separate, specialized areas whose activity also varied independently. We conclude that even in natural conditions, when many features have to be processed simultaneously, functional specialization is preserved. Our method thus opens a new way of brain mapping, which allows the localization of a multitude of brain areas based on a single experiment using uncontrolled, natural stimuli. Furthermore, our results show that the intensity of activity in a specialized area is linearly correlated with the intensity of its perceptual experience. This leads us to suggest that each specialized area is directly responsible for the creation of a feature specific conscious percept (a microconsciousness). Hum. Brain Mapp. 21:75-83, 2004. PMID- 14755596 TI - Effects of skull thickness, anisotropy, and inhomogeneity on forward EEG/ERP computations using a spherical three-dimensional resistor mesh model. AB - Bone thickness, anisotropy, and inhomogeneity have been reported to induce important variations in electroencephalogram (EEG) scalp potentials. To study this effect, we used an original three-dimensional (3-D) resistor mesh model described in spherical coordinates, consisting of 67,464 elements and 22,105 nodes arranged in 36 different concentric layers. After validation of the model by comparison with the analytic solution, potential variations induced by geometric and electrical skull modifications were investigated at the surface in the dipole plane and along the dipole axis, for several eccentricities and bone thicknesses. The resistor mesh permits one to obtain various configurations, as local modifications are introduced very easily. This has allowed several head models to be designed to study the effects of skull properties (thickness, anisotropy, and heterogeneity) on scalp surface potentials. Results show a decrease of potentials in bone, depending on bone thickness, and a very small decrease through the scalp layer. Nevertheless, similar scalp potentials can be obtained using either a thick scalp layer and a thin skull layer, and vice versa. It is thus important to take into account skull and scalp thicknesses, because the drop of potential in bone depends on both. The use of three different layers for skull instead of one leads to small differences in potential values and patterns. In contrast, the introduction of a hole in the skull highly increases the maximum potential value (by a factor of 11.5 in our case), because of the absence of potential drop in the corresponding volume. The inverse solution without any a priori knowledge indicates that the model with the hole gives the largest errors in both position and dipolar moment. Our results indicate that the resistor mesh model can be used as a robust and user-friendly simulation tool in EEG or event-related potentials. It makes it possible to build up real head models directly from anatomic magnetic resonance imaging without tessellation, and is able to take into account head heterogeneities very simply by changing volume elements conductivity. Hum. Brain Mapping 21:84-95, 2004. PMID- 14755597 TI - Behavioral conflict, anterior cingulate cortex, and experiment duration: implications of diverging data. AB - We investigated the relationship between behavioral measures of conflict and the degree of activity in the anterior cingulate cortex (ACC). We reanalyzed an existing data set that employed the Stroop task using functional magnetic resonance imaging [Milham et al., Brain Cogn 2002;49:277-296]. Although we found no changes in the behavioral measures of conflict from the first to the second half of task performance, we found a reliable reduction in the activity of the anterior cingulate cortex. This result suggests the lack of a strong relationship between behavioral measurements of conflict and anterior cingulate activity. A concomitant increase in dorsolateral prefrontal cortex activity was also found, which may reflect a tradeoff in the neural substrates involved in supporting conflict resolution, detection, or monitoring processes. A second analysis of the data revealed that the duration of an experiment can dramatically affect interpretations of the results, including the roles in which particular regions are thought to play in cognition. These results are discussed in relation to current conceptions of ACC's role in attentional control. In addition, we discuss the implication of our results with current conceptions of conflict and of its instantiation in the brain. Hum. Brain Mapping 21:96-105, 2004. PMID- 14755598 TI - Simple and complex movement-associated functional MRI changes in patients at presentation with clinically isolated syndromes suggestive of multiple sclerosis. AB - Using functional magnetic resonance imaging (fMRI), we investigated whether movement-associated functional changes of the brain are present in patients who are, most likely, at the earliest stage of multiple sclerosis (MS). Functional MRI exams were obtained from 16 patients at presentation with clinically isolated syndromes (CIS) suggestive of MS and 15 sex- and age-matched healthy volunteers during the performance of three simple and one more complex motor tasks with fully normal functioning extremities. fMRI analysis was performed using statistical parametric mapping (SPM99). Compared to healthy volunteers, CIS patients had increased activations of the contralateral primary sensorimotor cortex (SMC), secondary somatosensory cortex (SII), and inferior frontal gyrus (IFG), when performing a simple motor task with the dominant hand. The increased recruitment of the contralateral primary SMC was also found during the performance of the same motor task with the non-dominant hand and with the dominant foot. In this latter case, an anterior shift of the center of activation of this region was detected. During the performance of a complex motor task with the dominant upper and lower limbs, CIS patients had an increased recruitment of a widespread network (including the frontal lobe, the insula, the thalamus), usually considered to function in motor, sensory, and multimodal integration processing. The comparison of brain activations during the performance of simple vs. complex motor tasks showed that the movement-associated somatotopic organization of the cerebral and cerebellar cortices was retained in patients with CIS. Cortical reorganization occurs in patients at presentation with CIS highly suggestive of MS. Local synaptic reorganization, recruitment of parallel existing pathways, and reorganization of distant sites are all likely to contribute to the observed functional changes. Hum. Brain Mapping 21:106-115, 2004. PMID- 14755600 TI - Nanoanalysis by a high-resolution energy filtering transmission electron microscope. AB - An energy-filtering transmission electron microscope with 300 kV acceleration voltage was developed and the spatial resolution of elemental distribution images was improved. Observing oxygen monolayers in Al(11)O(3)N(9), it was shown that the actual resolution attained is up to 0.5 nm. Surface plasmon loss images of silver particles were taken with a resolution of better than 0.4 nm. Furthermore, the sensitivity is sufficiently high to distinguish indium content differences of 2.5 atomic percent in In(x)Al(1-x)As. This performance is good enough to analyze elemental distribution with atomic-level resolution. Furthermore, since analysis with the energy-filtering microscope is easy and practical, nanoanalysis may come into wide use not only in academic fields but also in industry. PMID- 14755599 TI - Automated detection and delineation of mitochondria in electron micrographs of human skeletal muscles. AB - Morphometric measurements of mitochondria in human skeletal muscles provide useful information relating to tissue oxidative energy production, nutrition, exercise, and aging. Morphometric data such as area, perimeter, long axis, and short axis can be obtained by delineating individual mitochondria in electron micrographs. However, manual counting and delineating of individual mitochondria is a formidable task. The purpose of this study was to develop a fully automated computer algorithm for quantifying mitochondrial morphometry in electron micrographs. The algorithm locates mitochondria with a two-dimensional matched filter and then traces the borders of individual mitochondria. The delineation is accomplished by edge detection along radial lines launched outwards from the center of each mitochondrion. Shape descriptors applied to delineated mitochondria are used to reject likely false-positive selections. The results show that the fully automated algorithm detects mitochondria with a false positive rate of 2% and a false-negative rate of 36%. The errors are easily and rapidly corrected by user intervention using a second semiautomated delineation algorithm. Morphometric measurements collected with the automated algorithm are equivalent to those obtained manually by human experts. The algorithm significantly improves the speed of image analysis and it also provides copious quantities of high-quality mitochondrial morphometric data. PMID- 14755601 TI - Perisynaptic Schwann cells of the vertebrate motor endplate bear modified cilia. AB - Perisynaptic Schwann cells (PSCs), descendants of the myelinating Schwann cells, cover the axon terminal of the vertebrate motor endplate of the skeletal muscle fiber. PSCs are assumed to support the function of the axon terminal. This function suggests a net material transport in the direction of the axon terminal. Morphologically it is to be expected that these cells have a cytoskeleton aligned to the axon terminal. Investigations clarifying this statement have not yet been undertaken. From previous investigations we know, however, that the PSCs have a microtubule-organizing center, which is a part of this cytoskeleton. The centrioles of the organizing center may also participate in the formation of a modified cilium structure whose function is unknown. In the present investigation, characteristic ultrastructural features of the modified cilium structure and its relationship to the Golgi apparatus and the axon terminal are presented. A function for the modified cilium structure is discussed. PMID- 14755602 TI - Ultrastructural features of bone marrow cells from patients with acquired sideroblastic anemia. AB - The ultrastructural findings of the bone marrow cells from 15 patients with acquired sideroblastic anemia are presented. The red cell precursors from all patients showed the presence of electron-dense material in the mitochondria, representing most probably iron deposits. A great number of these mitochondria were completely destroyed. The erythropoietic precursors from one of the patients showed markedly elongated mitochondria that measured up to 3 microm. In addition numerous cytoplasmic vacuoles were observed. The red cell precursors from 60% of the patients showed signs of dyserythropoiesis, such as incomplete nuclear division and nuclear distortion. The polymorphonuclears from 47% of the patients presented nuclear abnormalities expressed as nuclear bridges, appendices, and blebs. In addition, phagocytosis of red blood cells was observed. The results of the study underline the advantages of the transmission electron microscope examination in visualization of intricate alterations in hematopoietic cells that cannot be detected with a light microscope. PMID- 14755603 TI - Visualization of vesicle transport along and between distinct pathways in neurites of living cells. AB - Trafficking of secretory vesicles along neurites of PC12 cells was visualized by 2D and 3D real-time imaging using fluorescence microscopy. Vesicle motion along distinct pathways was directly seen. From an overlay of individual pathways, the underlying cytoskeletal filament could be imaged at a subwavelength resolution. Continuous vesicle transport was interrupted by periods of diffusive motion with concomitant pathway changes. Statistical analysis shows that such interruptions were distributed stochastically along the filament, indicating a limited processivity of motor proteins also in a cellular context. Periods of diffusive motion facilitated the interaction with actively transported vesicles. Frequent associations and dissociations of vesicles have been observed consistently, pointing to a functional relevance of vesicle cotransport. PMID- 14755604 TI - Comparison of structural and hemostatic properties of the poly-N-acetyl glucosamine Syvek Patch with products containing chitosan. AB - Polysaccharides are becoming increasingly developed as therapeutics and medical products, as the new field of Glycomics expands. Glycosaminoglycans that contain N-acetyl glucosamine constituents have been the focus of research leading to medical devices. A new hemostatic bandage, the Syvek Patch, has been introduced in the recent past for the control of bleeding at vascular access sites in interventional cardiology and radiology procedures. This product consists of poly N-acetyl glucosamine (pGlcNAc) isolated in a unique fiber crystalline structural form from the large-scale culture and processing of a marine diatom. The Syvek pGlcNAc fiber material has chemical, physical, and biological properties that result in its favorable performance as a hemostat. Two new products, the Clo-Sur PAD and ChitoSeal, have recently become available also as patch hemostats. These two products both use chitosan, another N-acetyl glucosamine containing glycosaminoglycan, as their active ingredient. Structural, chemical, and biological comparisons of Syvek pGlcNAc and chitosan reveal a number of important differences. Syvek pGlcNAc fibers contain approximately 50 fully acetylated, high molecular weight pGlcNAc molecules in a crystalline, three-dimensional beta structure array, and are insoluble. Chitosan is a low molecular weight mixed amorphous cationic polymer with no regular structure as a solid, and is water soluble taking on a random coil configuration when in solution. These structural dissimilarities result in differences in the hemostatic properties of the two materials. Syvek pGlcNAc is able to significantly reduce the in vitro fibrin clot formation time of platelet-rich plasma samples and has the ability to cause aggregation of red blood cells in vitro. Chitosan is no better than gauze or other controls in these in vitro assays. The Syvek Patch is able to control the bleeding and cause hemostasis in a coagulopathic swine spleen-bleeding animal model 100% of the time, whereas Clo-Sur PAD was completely unsuccessful (0%) and ChitoSeal (25%) was worse than a gauze pad control (50%) in the same model. Syvek pGlcNAc fibers have structural and chemical properties that provide a unique basis for their ability to interact with blood components to cause hemostasis. Chitosan does not have the same properties and capabilities. PMID- 14755605 TI - The use of optical parametric oscillator for harmonic generation and two-photon UV fluorescence microscopy. AB - Ultrafast lasers have found increasing use in scanning optical microscopy due to their very high peak power in generating multiphoton excitations. A mode-locked Ti:sapphire laser is often employed for such purposes. Together with a synchronously pumped optical parametric oscillator (OPO), the spectral range available can be extended to 1,050-1,300 nm. This broader range available greatly facilitates the excitation of second harmonic generation (SHG) and third harmonic generation (THG) due to better satisfaction of phase matching condition that is achieved with a longer excitation wavelength. Dental sections are then investigated with the contrasts from harmonic generation. In addition, through intra-cavity doubling wavelengths from 525-650 nm are made available for effective two-photon (2-p) excitation with the equivalent photon energy in the UVB range (290-320 nm) and beyond. This new capacity allows UV (auto-) fluorescence excitation and imaging, for example, from some amino acids, such as tyrosine, tryptophan, and glycine. PMID- 14755606 TI - Matrix-assisted laser desorption/ionization mass spectrometry for the evaluation of the C-terminal lysine distribution of a recombinant monoclonal antibody. AB - Recombinant monoclonal antibodies produced using mammalian cell lines contain multiple chemical modifications. One specific modification resides on the C terminus of the heavy chain. Enzymes inside the cell can cleave the C-terminal lysine from the heavy-chain molecules, and variants with and without C-terminal lysine can be produced. In order to fully characterize the protein, there is a need for analytical methods that are able to account for the different product variants. Conventional analytical methods used for the measurement of the distribution of the two different variants are based on chemical or enzymatic degradation of the protein followed by chromatographic separation of the degradation products. Chromatographic separations with gradient elution have long run times, and analyses of multiple samples are time-consuming. This paper reports development of a novel method for the determination of the relative amounts of the two C-terminal heavy-chain variants based on matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) measurements of the cyanogen bromide degraded recombinant monoclonal antibody products. The distribution of the variants is determined from the MALDI-TOF mass spectra by measuring the peak areas of the two C-terminal peptides. The assay was used for the assessment of the C-terminal lysine distribution in different development lots. The method was able to differentiate between the products obtained using the same cell line as well as between products obtained from different cell lines. PMID- 14755607 TI - Using dual-bacterial denitrification to improve delta15N determinations of nitrates containing mass-independent 17O. AB - The bacterial denitrification method for isotopic analysis of nitrate using N(2)O generated from Pseudomonas aureofaciens may overestimate delta(15)N values by as much as 1-2 per thousand for samples containing atmospheric nitrate because of mass-independent (17)O variations in such samples. By analyzing such samples for delta(15)N and delta(18)O using the denitrifier Pseudomonas chlororaphis, one obtains nearly correct delta(15)N values because oxygen in N(2)O generated by P. chlororaphis is primarily derived from H(2)O. The difference between the apparent delta(15)N value determined with P. aureofaciens and that determined with P. chlororaphis, assuming mass-dependent oxygen isotopic fractionation, reflects the amount of mass-independent (17)O in a nitrate sample. By interspersing nitrate isotopic reference materials having substantially different delta(18)O values with samples, one can normalize oxygen isotope ratios and determine the fractions of oxygen in N(2)O derived from the nitrate and from water with each denitrifier. This information can be used to improve delta(15)N values of nitrates having excess (17)O. The same analyses also yield estimates of the magnitude of (17)O excess in the nitrate (expressed as Delta(17)O) that may be useful in some environmental studies. The 1-sigma uncertainties of delta(15)N, delta(18)O and Delta(17)O measurements are +/-0.2, +/-0.3 and +/-5 per thousand, respectively. PMID- 14755608 TI - Analysis of derivatized and underivatized theanine enantiomers by high performance liquid chromatography/atmospheric pressure ionization-mass spectrometry. AB - Theanine, a naturally occurring non-proteinic amino acid found in tea leaves, has demonstrated wide-ranging physiological activity, from lowering blood pressure to enhancing the anti-tumor activity of chemotherapeutic drugs. The chiral nature of theanine suggests that enantiospecificity plays a significant role in its various pharmacological functions. Using the Chirobiotic T (teicoplanin) chiral stationary phase, native and derivatized theanine enantiomers were separated and detected via high-performance liquid chromatography (HPLC) coupled to atmospheric pressure ionization mass spectrometry (API-MS). With the use of flow rates compatible with each ionization source, native theanine standards achieved excellent sensitivity and detection limits (10 ng/mL) for both atmospheric pressure chemical ionization (APCI) and electrospray ionization (ESI). Optimum sensitivity and detection limits for derivatized theanine standards were achieved using ESI-MS. The enantiomeric composition of six commercially available L theanine samples was evaluated using the high-flow APCI-MS method and confirmed with photodiode array detection. Five of the six products contained significant amounts of D-theanine. Only one product, SunTheanine, appeared to contain only the L-theanine enantiomer. PMID- 14755609 TI - Comparison of mono- and polyatomic primary ions for the characterization of organic dye overlayers with static secondary ion mass spectrometry. AB - Organic carbocyanine dye coatings have been analyzed by time-of-flight static secondary ion mass spectrometry (TOF-S-SIMS) using three types of primary ions: Ga(+) operating at 25 keV, and Xe(+) and SF(5) (+) both operating at 9 keV. Secondary ion yields obtained with these three primary ions have been compared for coatings with different layer thickness, varying from (sub)-monolayer to multilayers, on different substrates (Si, Ag and AgBr cubic microcrystals). For (sub)-monolayers deposited on Ag, Xe(+) and SF(5) (+) primary ions generate similar precursor ion intensities, but with Ga(+) slightly lower precursor ion intensities were obtained. Thick coatings on Ag as well as mono- and multilayers on Si produce the highest precursor and fragment ion intensities with the polyatomic primary ion. The yield difference between SF(5) (+) and Xe(+) can reach a factor of 6. In comparison with Ga(+), yield enhancements by up to a factor of 180 are observed with SF(5) (+). For the mass spectrometric analysis of dye layers on AgBr microcrystals, SF(5) (+) again proves to be the primary ion of choice. PMID- 14755610 TI - Simultaneous determination of the urinary metabolites of benzene, toluene, xylene and styrene using high-performance liquid chromatography/hybrid quadrupole time of-flight mass spectrometry. AB - A simple and rapid method using reversed-phase liquid chromatography/tandem mass spectrometry (LC/MS/MS) for the simultaneous determination of the urinary metabolites of benzene, toluene, xylene and styrene in human urine specimens and standard solutions is described. A hybrid quadrupole/time-of-flight (QqTOF) mass spectrometer was compared for the determination of metabolite of aromatic solvents in urine samples. The metabolites selected were: trans,trans-muconic acid, hippuric acid, o-, m- and p-methylhippuric acid and phenylglyoxylic acid. The compounds were well separated from each other on narrow-bore 1-mm i.d. reversed-phase LC C-18 columns. Average recoveries for loading 100 microL of urine samples varied from 88-110% and the quantification limits were less than 30 ng/mL for each analyte (3 ng/mL for trans,trans-muconic acid). The qualitative information obtained (mass accuracy, resolution and full-scan spectra) with the QqTOF mass spectrometer allows a secure identification of analytes in biological matrices. PMID- 14755611 TI - Synthesis and electrospray ionization mass spectra of AZT/d4T boranophosphates. AB - New anti-HIV prodrugs, conjugates of AZT and d4T with boranophosphates, were prepared by the H-phosphonate method. Their structures were determined by negative ion electrospray ionization mass spectrometry (ESI-MS) in conjunction with tandem mass spectrometry (MS/MS). The fragmentation pathways were investigated, and most of the fragment ions contained the boranophosphate or phosphinate group. PMID- 14755612 TI - Surface chemistry study of RuO2/IrO2/TiO2 mixed-oxide electrodes. AB - DSA metal oxide electrodes such as the RuO(2)/IrO(2)/TiO(2) mixed system are widely studied for their excellent electrocatalytic activity. In order to understand their catalytic properties, the comprehension of the surface chemistry involved during electrochemical treatments is crucial. With this aim, RuO(2)/IrO(2)/TiO(2) mixed-oxide electrodes having various noble metal contents were studied by means of secondary ion mass spectrometry (SIMS). In particular, cathodic and anodic polarization and O(2) evolution reactions were carried out to test the electrode behaviour and SIMS analyses were performed after all these treatments. In this way, surface changes induced by electrochemical treatments and depending on electrode composition were widely investigated by SIMS, revealing, for example, the presence of hydration or preferential dissolution phenomena induced by electrochemical processing. PMID- 14755613 TI - Increasing throughput of parallel on-line extraction liquid chromatography/electrospray ionization tandem mass spectrometry system for GLP quantitative bioanalysis in drug development. AB - An approach is described with turbulent flow on-line extraction liquid chromatography/electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) for GLP quantitative bioanalysis of a drug candidate. Two systems were built in house with standard laboratory parts and equipments. One system consisted of one gradient HPLC pump, one isocratic pump, one ten-port valve, two turbulent flow columns, one analytical column, one autosampler and one mass spectrometer. Using this system, an injection-to-injection cycle time of 0.8 min was achieved. By adding an additional valve, another analytical column and an isocratic pump, the injection-to-injection cycle time decreased to 0.4 min. Validation results from the two systems showed that precision and accuracy were acceptable for GLP quantitative analyses. The system was utilized to support sample bioanalysis of a drug candidate in a first-time in-human clinical trial. PMID- 14755614 TI - Quantitative determination of the polypeptide motilin in rat plasma by externally calibrated liquid chromatography/electrospray ionization mass spectrometry. AB - We present a method for the quantitation of motilin from rat plasma by protein precipitation and liquid chromatography/mass spectrometry (LC/MS). Using external calibration, the method was linear over the concentration range 10-1000 ng/mL with an initial sample volume of 150 microL. The LC system included a C(18) column with a 300 A pore size. A linear gradient was used with a mobile phase consisting of water and acetonitrile, each with 0.2% acetic acid and 0.02% trifluoroacetic acid. Motilin was detected with the mass spectrometer in positive ion mode monitoring the 4+ charge state at m/z 675.5. The approximated limit of detection was less than 1 ng/mL and the lower limit of quantitation (LLOQ) was 10 ng/mL. The method showed a high degree of precision and accuracy both within and between runs at five validation points, including the LLOQ. PMID- 14755615 TI - Continuous-flow isotope analysis of the deuterium/hydrogen ratio in atmospheric hydrogen. AB - A convenient method is described for analyzing the deuterium/hydrogen (D/H) ratio of atmospheric molecular hydrogen (H(2)) based on mass spectrometric isotope ratio monitoring. The method requires small amounts of air ( approximately 300 mL STP), is operated on-line, and comprises four steps: (1). the condensation of the air matrix at approximately 40 K; (2). the collection of the non-condensed components of the air sample (H(2), Ne, He, and traces of N(2)) in a 5 A molecular sieves pre-concentration trap at approximately 63 K; (3). gas chromatographic purification of H(2) in a flow of He; and (4) quantification of the D/H ratio in an isotope-ratio mass spectrometer. The precision of the determination of the D/H ratio is better than 2 per thousand, which is comparable to, or better than, that obtained by conventional duel-inlet off-line analysis. There are, however, discrepancies relative to the D/H ratios determined by conventional duel-inlet analysis. This is due to differences in peak shape between reference and sample air, depending on the amount of H(2) injected. Consequently, calibration runs are required. After the calibration of the system, we obtained an accuracy of 1.5 per thousand, so that the accumulated uncertainty is estimated to be less than 4 per thousand. The method also allows determination of the H(2) concentration, with an uncertainty estimated to be 2%. PMID- 14755616 TI - Ab initio calculations of proton affinities of glycine, proline, cysteine and phenylalanine: comparison with the experimental values obtained using an electrospray ionisation ion trap mass spectrometer. AB - The proton affinities of four characteristic amino acids, glycine, proline, cysteine and phenylalanine, have been calculated using three different types of wave function, HF, DFT and MP2. These wave functions were combined with several basis sets using the valence double- or triple-xi with polarisation functions, and included or not a diffuse d function on heavy atoms. Calculations were carried out using Gaussian 98 on a LINUX system (2 GHz, 2 GB of RAM). The calculated results have been compared with the experimental values obtained using Cooks' kinetic method, in particular on an electrospray ionisation ion trap mass spectrometer. An excellent agreement was found between the experimental values and the theoretical results obtained using the B3P86/6-31+G*//B3LYP/6-31G* level. PMID- 14755617 TI - Glass-chip-based sample preparation and on-chip trypic digestion for matrix assisted laser desorption/ionization mass spectrometric analysis using a sol gel/2,5-dihydroxybenzoic acid hybrid matrix. AB - A glass-chip-based sample preparation method for matrix-assisted laser desorption/ionization mass spectrometric (MALDI-MS) analysis of tryptic digests of proteins and intact cells is described. A MALDI matrix, 2,5-dihydroxybenzoic acid (2,5-DHB), was hybridized with sol-gels to generate a sol-gel-derived material. Taking advantage of the characteristics of sol-gels, the sol-gel derived material readily adhered to the surface of a glass chip through covalent bonding. Only one step of sample preparation, deposition of the sample solution on the glass chip, was required before MALDI-MS analysis. Because 2,5-DHB was homogeneously dispersed on the sol-gel network structure, good spot-to-spot reproducibility was obtained in MALDI analysis using this approach and the analyte signals were uniform throughout the chip. The modified glass chips were robust and effective for at least 1 week. This glass-chip-based matrix preparation method provides a straightforward approach to developing techniques for analyzing the on-chip enzymatic digestion of proteins and intact cells of microorganisms. Cytochrome C and Escherichia coli were used as analytes to demonstrate the feasibility of this approach. The products of the on-chip enzymatic digests were identified through protein database searches. PMID- 14755618 TI - Rapid screening for S-adenosylmethionine-dependent methylation products by enzyme transferred isotope patterns analysis. AB - We report here an isotopic labeling and mass spectrometric method to rapidly identify S-adenosylmethionine (AdoMet)-dependent methylation products. In the presence of CH(3)- and CD(3)-labeled AdoMet, a methyl transfer product appears as a doublet separated by 3 Da in a mass spectrum, while other compounds show their normal isotopic distribution. Based on this unique isotopic pattern, methylation product(s) can be easily detected even from a mixture of cellular components. To validate our method, the product of human thiopurine methyltransferase (TPMT, EC 2.1.1.67) has been successfully identified from both an in vitro assay and a whole-cell assay. This method is generally applicable to AdoMet-dependent transmethylation and other group-transfer reactions, and constitutes the first example of a general strategy of enzyme-transferred isotope patterns (ETIPs) analysis. PMID- 14755619 TI - [2H/H] Isotope ratio analyses of [2H5]cholesterol using high-temperature conversion elemental analyser isotope-ratio mass spectrometry: determination of cholesterol absorption in normocholesterolemic volunteers. AB - This paper validates the use of high-temperature conversion elemental analyser isotope-ratio mass spectrometry (TC-EA/IRMS) for measuring the [(2)H/H] enrichment of plasma [(2)H(5)]cholesterol. From a molecular point of view, the free cholesterol is initially separated from plasma by thin-layer chromatography (TLC) and then injected onto the TC-EA reactor which converts cholesterol molecules into CO and H(2) gases. The slope of the curve of the experimental mole percent excess (MPE((exp.))) versus MPE((theor.)) was very close to 1, demonstrating that no significant isotopic fractionation was observed during all processing of the samples (i.e., isolation of plasma free cholesterol by TLC and pyrolysis in the TC-EA reactor). Excellent linearity (r(2) = 0.9994, n = 4) of delta ( per thousand ) of [(2)H/H] isotopic measurements versus mole percent (MP) was assessed over the range 0 to 0.1 MP. The precision of the [(2)H/H] measurement, evaluated with two calibration points processed with TLC, was delta(2)H(V-SMOW) = -192.5 +/- 3.4 per thousand and delta(2)H(V-SMOW) = -136.9 +/ 2.9 per thousand. The standard deviations of the within-assay and between-assay repeatabilities of the analytical process, evaluated using the quality control (QC) of plasma samples, were 4.6 and 6.1 per thousand, respectively. Plant sterols are known to reduce cholesterol absorption and therefore were used as a positive control in a clinical study performed with normocholesterolemic volunteers. This present method produces biological results consistent with those already reported in the literature. PMID- 14755620 TI - Oligomeric carbon and siloxane series observed by matrix-assisted laser desorption/ionisation and laser desorption/ionisation mass spectrometry during the analysis of soot formed in fuel-rich flames. AB - Oligomeric carbon and siloxane series have been observed by matrix-assisted laser desorption/ionisation mass spectrometry (MALDI-MS), during the analysis of the dichloromethane (DCM)-soluble fractions of condensable material recovered from fuel-rich flames. Laser desorption (LD) spectra showed a pattern of oligomeric dimethyl-siloxane structures with a spacing of 74 u. The siloxane series appears to have originated as contamination of samples by silicone oil used to lubricate connections of polymer tubing. This was confirmed by extracting silicone tubing and silicone grease with DCM followed by MALDI-MS analysis. A series of peaks with a mass spacing of 24 u was also observed, superimposed on the continuum of unresolved organic ions. This oligomeric series appears to correspond to polycyclic aromatics separated by (mainly) ethylene bridges. Thus LD-MS appears to have revealed a series of soot precursors, intermediate between polycyclic aromatics and particulate soot, which was not detected by MALDI-MS. More detailed work is necessary to define these species with precision. PMID- 14755621 TI - Advantages of derivatization by osmium tetroxide and 2,2'-bipyridine for matrix assisted laser desorption/ionization post-source decay fragment ion analysis of peptides. AB - Matrix-assisted laser desorption/ionization--post-source decay (MALDI-PSD) fragment ion analysis is frequently used for peptide sequence determination. PSD fragmentation is often changed or improved in terms of, e.g., sequence coverage, after derivatization. In this work, the influence of modification by an osmium tetroxide-bipyridine reagent (Os,bipy) on the MALDI-PSD behaviour of peptides is studied. The reagent modifies peptides specifically at tryptophan residues and oxidizes methionine to methionine sulfone and cysteine to cysteic acid. As a result the masses of some of the fragments are specifically shifted in case of peptides containing a methionine by +32 Da and, in cases of peptides containing a cysteine residue, by +48 Da. In addition, due to the change in protonation properties of a peptide after oxidation, fragments containing cysteic acid are in most cases totally suppressed. This effect significantly facilitates peptide sequence determination. Improvement of MALDI-TOFMS and PSD analysis after the reaction with Os,bipy is demonstrated for examples involving derivatives of humanin, a novel neuroprotective peptide. PMID- 14755622 TI - Experimental validation of theoretical potassium and sodium cation affinities of amides by mass spectrometric kinetic method measurements. AB - In this study the theoretical Gaussian-2 K(+)/Na(+) binding affinities (enthalpies) at 0 K (in kJ mol(-1)) for six amides in the order: formamide (109.2/138.5) < N-methylformamide (117.7/148.6) < acetamide (118.7/149.5) < N,N dimethylformamide (123.9/156.4) < N-methylacetamide (125.6/157.7) < N,N dimethylacetamide (129.2/162.6), reported previously (Siu et al., J. Chem. Phys. 2001; 114: 7045-7051), were validated experimentally by mass spectrometric kinetic method measurements. By monitoring the collision-induced dissociation (CID) of K(+)/Na(+)-bound heterodimers of the amides, the relative affinities were shown to be accurate to within +/-2 kJ mol(-1). With these six theoretical K(+)/Na(+) binding affinities as reference values, the absolute K(+)/Na(+) affinities of imidazole, 1-methylimidazole, pyridazine and 1,2-dimethoxyethane were determined by the extended kinetic method, and found to be consistent (to within +/-9 kJ mol(-1)) with literature experimental values obtained by threshold CID, equilibrium high-pressure mass spectrometry, and Fourier transform ion cyclotron resonance/ligand-exchange equilibrium methods. A self-consistent resolution is proposed for the inconsistencies in the relative order of K(+)/Na(+) affinities of amides reported in the literature. These two sets of validated K(+) and Na(+) affinity values are useful as reference values in kinetic method measurements of K(+)/Na(+) affinity of model biological ligands, such as the K(+) affinities of aliphatic amino acids. PMID- 14755623 TI - Formation of [M-H]- and [M-2H]2- ions in the electrospray ionization mass spectra of dicarboxylated polyethylene glycols. AB - The negative-ion electrospray mass spectrometric behavior of dicarboxylated polyethylene glycols (CPEGCs) is discussed. Both [M-H](-) and [M-2H](2-) ions were observed. It was found that the ratio [M-2H](2-)/[M-H](-) is affected by oxyethylene chain length, solvent polarity, analyte concentration and applied cone voltage. PMID- 14755624 TI - Application of 9-nitroanthracene as a matrix for laser desorption/ionization analysis of fluorinated fullerenes. PMID- 14755625 TI - Electron ionization mass spectrometry in the characterization of azidonitriles. PMID- 14755626 TI - Efficient matrix-assisted laser desorption/ionization method for fully pi conjugated dendrimers. PMID- 14755627 TI - Effects of acute acetylcholinesterase inhibition on the cerebral cholinergic neuronal system and cognitive function: Functional imaging of the conscious monkey brain using animal PET in combination with microdialysis. AB - This study demonstrated the effects of acute acetylcholinesterase (AChE) inhibition by donepezil (Aricept) on the cerebral cholinergic neuronal system in the brains of young (5.2 +/- 1.1 years old) and aged (20.3 +/- 2.6 years old) monkeys (Macaca mulatta) in the conscious state. Donepezil at doses of 50 and 250 microg/kg suppressed AChE activity, analyzed by metabolic rate (k(3)) of N [(11)C]methyl-4-piperidyl acetate ([(11)C]MP4A), in all cortical regions in a dose-dependent manner in both age groups. However, the suppression degree was more marked in young than in aged monkeys. AChE inhibition by donepezil resulted in a dose-dependent increase in acetylcholine levels in the prefrontal cortex of young animals as measured by microdialysis. Binding of (+)N-[(11)C]propyl-3 piperidyl benzilate ([(11)C](+)3-PPB) to cortical muscarinic receptors was reduced by donepezil, probably in a competitive inhibition manner. Aged monkeys showed less reduction of [(11)C](+)3-PPB binding than young animals. As evaluated by an oculomotor delayed response task, aged monkeys showed impaired working memory performance compared to young monkeys, and the impaired performance was partly improved by the administration of donepezil, due to the facilitation of the cholinergic neuronal system by AChE inhibition. These results demonstrate that the PET imaging technique with specific labeled compounds in combination with microdialysis and a behavioral cognition task could be a useful method to clarify the mechanism of drugs in the living brains of experimental animals. PMID- 14755628 TI - Methamphetamine-induced loss of striatal dopamine innervation in BDNF heterozygote mice does not further reduce D3 receptor concentrations. AB - Depletion of dopamine (DA) reduces D(3) receptor number, but D(3) receptor expression is also regulated by brain-derived neurotrophic factor (BDNF). We took advantage of transgenic heterozygous BDNF mutant mice (+/-) to determine if reduced BDNF and loss of DA fibers produced by methamphetamine were additive in their impact on D(3) receptor number. We assessed selective markers of the dopaminergic system including caudate-putamen DA concentrations and quantitative autoradiographic measurement of tyrosine hydroxylase (TH) levels, DA transporter (DAT), and DA D(3) receptor binding between vehicle and methamphetamine-treated BDNF +/- and their wildtype (WT) littermate control mice. Caudate-putamen DA concentrations, TH and DAT levels were significantly reduced following methamphetamine treatment in both WT and BDNF +/- mice. The extent of methamphetamine-induced reduction in TH and DAT was greater for the WT than BDNF +/- mice and DAT levels were also decreased to a greater extent in nucleus accumbens of WT as compared to BDNF +/- mice. Lower D(3) receptor existed in caudate-putamen and nucleus accumbens in BDNF +/- mice and these differences were not affected by methamphetamine treatment. Taken together, these results not only substantiate the importance of BDNF in controlling D(3) receptor expression, but also indicate that a methamphetamine-induced depletion of DA fibers fails to produce an additive effect with lowered BDNF for control of D(3) receptor expression. In addition, the reduction of D(3) receptor expression is associated with a decreased neurotoxic response to methamphetamine in BDNF +/- mice. PMID- 14755629 TI - Effect of the acute and chronic administration of the selective 5-HT6 receptor antagonist SB-271046 on the activity of midbrain dopamine neurons in rats: an in vivo electrophysiological study. AB - This study examined the effect of the acute and repeated per os (p.o.) administration of the selective 5-HT(6) receptor antagonist SB-271046, on the number, as well as the firing pattern of spontaneously active dopamine (DA) neurons in the rat substantia nigra pars compacta (SNC) and ventral tegmental area (VTA) in anesthetized male Sprague-Dawley rats. This was accomplished using the technique of extracellular in vivo electrophysiology. A single p.o. administration of either 1, 3, or 10 mg/kg of SB-271046 did not significantly alter the number of spontaneously active SNC DA neurons per stereotaxic electrode tract compared to vehicle-treated animals. The acute administration of either 1 or 3 mg/kg of SB-271046 did not significantly alter the number of spontaneously active VTA DA neurons. In contrast, a significant decrease in the number of spontaneously active VTA DA neurons was observed after a single administration of 10 mg/kg of SB-271046 compared to vehicle-treated animals. The acute p.o. administration of SB-271046 significantly altered the firing pattern parameters of all (bursting + nonbursting DA neurons) DA neurons, particularly those in the VTA, compared to vehicle-treated animals. The repeated p.o. administration (once per day for 21 days) of 1, 3, or 10 mg/kg of SB-271046 did not significantly alter the number of spontaneously active VTA DA neurons compared to vehicle treated animals. The repeated administration of 3 or 10 mg/kg of SB-271046 significantly increased the number of spontaneously active SNC DA neurons compared to vehicle controls. Overall, the repeated administration of SB-271046 had relatively little effect on the firing pattern of midbrain DA neurons. The results obtained following the chronic administration of SB-271046 show that this compound has a profile different from that of typical or atypical antipsychotic drugs in this model. Clinical studies are required to understand what role 5 HT(6) receptor blockade might eventually play in the treatment of schizophrenia. PMID- 14755630 TI - Evaluation of [18F]fluorinated sigma receptor ligands in the conscious monkey brain. AB - PET-imaging of the sigma receptors is very helpful to understand processes, e.g., several central nervous system (CNS)-diseases in which the sigma receptors are involved. The [(18)F]fluoroethylated analogs of SA4503 and SA5845 ([(18)F]FE SA4503 and [(18)F]FE-SA5845) were evaluated in conscious monkeys to estimate its suitability for human application for PET. Conscious monkeys (Macaca Mulatta) were either scanned with [(18)F]FE-SA4503 or [(18)F]FE-SA5845 (n = 3 for both groups, 220-802 MBq). After a dynamic study of 120 min, radioactivity was displaced by intravenous (i.v.) injection of haloperidol (1 mg/kg). One month later the same set of three monkeys were scanned with [(18)F]FE-SA4503 for 120 min and "cold" SA4503 (1 mg/kg) was infused to displace the radioactivity, and the other three monkeys were pretreated with haloperidol (1 mg/kg) before the 120 min PET-scan with [(18)F]FE-SA5845. Cortical areas (cingulate, frontal, occipital, parietal, temporal), striatum, and thalamus showed high radioactivity uptake. Infusion of haloperidol displaced the radioactivity levels of the two radioligands. The same effect was found for [(18)F]FE-SA4503 after SA4503 displacement. Pretreatment with haloperidol blocked the [(18)F]FE-SA5845 binding to give PET-images with low and uniform uptake in the brain. The findings demonstrated the reversible binding of the two radioligands. Metabolite analysis showed that 14% and 23% parent compound of [(18)F]FE-SA5845 and [(18)F]FE-SA4503, respectively, at 120 min postinjection was present in plasma. Kinetic analysis showed that the binding potential of [(18)F]FE-SA5845 was higher in all brain regions than that of [(18)F]FE-SA4503 (4.75-8.79 vs. 1.65-4.04). The highest binding potential was found in the hippocampus, followed by the cortical regions, thalamus, cerebellar hemisphere, striatum and vermis. Both [(18)F]FE-SA compounds bound specifically to cerebral sigma receptors of the monkey and have potential for mapping sigma receptors in the human brain. PMID- 14755631 TI - Ultrastructural localization of Leu5-enkephalin immunoreactivity in mesocortical neurons and their input terminals in rat ventral tegmental area. AB - Enkephalin (ENK) immunoreactivity is widely distributed in the ventral tegmental area (VTA), where endogenous ENK and dynorphin opioid peptides are known to have opposing actions in reward, stress, cognition, and fear-related behaviors. Many neurons in the VTA give rise to mesocortical projections terminating in the medial prefrontal cortex (mPFC), and these projections have been implicated to varying extents in all these functions. To determine whether there is a synaptic basis for ENK and/or dynorphin modulation of cortically projecting neurons within the VTA, we combined retrograde tract-tracing from the mPFC with dual immunocytochemical-labeling electron microscopy in the rat VTA. The retrograde tracer Fluorogold (FG) was microinjected into mPFC. At optimal survival periods, sections through the VTA were processed for immunolabeling of anti-FG and a Leu(5)-ENK antibody recognizing both ENK and dynorphin peptides. Over 26% of the retrogradely labeled neuronal somatodendritic profiles (n = 177) were contacted by ENK-immunoreactive axonal profiles including small axons and axon terminals. The axon terminals varied in their subcellular distribution of ENK immunoreactivity and also differed in forming either inhibitory-type (symmetric) or excitatory-type (asymmetric) synapses. Many of the axonal profiles also were apposed to FG-labeled somata or dendrites without forming recognizable synapses. Approximately one-third of the mesocortical neuronal perikarya also showed sparsely distributed somatodendritic ENK-immunoreactivity. Our results provide ultrastructural evidence that ENK and possibly dynorphin in the rat VTA have distributions consistent with involvement in diverse physiological actions affecting the output of mesocortical neurons, some of which also contain one or both peptides. PMID- 14755632 TI - Aging impairs the late phase of long-term potentiation at the medial perforant path-CA3 synapse in awake rats. AB - The effects of aging on long-term potentiation (LTP) in the dentate gyrus (DG) and CA1 are well documented, but LTP at the medial perforant path (MPP)-CA3 synapse of aged animals has remained unexplored. Because the MPP-DG and Schaffer collateral-CA1 synapses account for only about 20% of total hippocampal synapses, global understanding of how aging affects hippocampal plasticity has remained limited. Much is known about LTP induction in the hippocampal formation, whereas the mechanisms that regulate LTP maintenance are less understood, especially during aging. We investigated the effects of aging on MPP-CA3 LTP induction and maintenance in awake rats. As is the case in the DG and CA1, high-frequency stimulation-induced LTP at the MPP-CA3 synapse is normal in aged rats. These data indicate that N-methyl-D-aspartate (NMDA) receptor-mediated processes are intact at the MPP-CA3 synapse in aged rats. In contrast, aging impaired the magnitude and duration of MPP-CA3 LTP over a period of days. Also, these data are consistent with reports that area CA3 is especially susceptible to age-related changes. Our data suggest that aging impairs mechanisms that regulate the late phase of MPP-CA3 LTP and contribute to a more global understanding of how aging affects hippocampal plasticity. PMID- 14755633 TI - Effects of 6-cyano-7-nitroquinoxaline-2,3-dione on nicotinic receptor subunit transcript expression in the rat brain. AB - The nicotinic cholinergic system exerts potent modulatory effects on glutamatergic neurotransmission, an effect mediated in part by increased glutamate release following activation of presynaptic nicotinic cholinergic receptors. Ionotropic glutamate receptor agonists also stimulate release of acetylcholine, suggesting that these neurotransmitter systems reciprocally regulate one another. We investigated an interface between the nicotinic cholinergic and glutamatergic systems by measuring nicotinic receptor subunit transcript expression following administration of 6-cyano-7-nitroquinoxaline-2,3 dione (CNQX), an antagonist of the AMPA and kainate subtypes of glutamate receptors. Using [(35)S] in situ hybridization, we measured expression of alpha 2, alpha 3, alpha 4, alpha 5, alpha 7, beta 2, beta 3, and beta 4 nicotinic receptor subunit transcripts in the rat forebrain. Following 7 days of treatment with vehicle or CNQX (1 mg/kg/day or 10 mg/kg/day), changes in nicotinic receptor subunit transcript expression were restricted to subunits that form heteromeric receptors. We found increased levels of transcripts for alpha 2 and beta 2 nicotinic receptor subunits in the hippocampus, decreased alpha 4 subunit transcripts in the medial habenula and amygdala, and increased beta 2 subunit transcripts in the septum and piriform cortex. We did not detect changes in expression of transcripts for the alpha 7 subunit, which forms homomeric nicotinic receptors. Our findings indicate that expression of nicotinic cholinergic receptor subunit transcripts are regulated in a subunit- and region specific fashion by CNQX, an antagonist of non-NMDA ionotropic glutamate receptors. PMID- 14755634 TI - The highly selective 5-hydroxytryptamine (5-HT)2A receptor antagonist, EMD 281014, significantly increases swimming and decreases immobility in male congenital learned helpless rats in the forced swim test. AB - We examined the effect of the highly selective 5-hydroxytryptamine (5-HT)(2A) receptor antagonist 7-[4-[2-(4-fluoro-phenyl)-ethyl]-piperazine-1-carbonyl]-1H indole-3-carbonitrile HCl (EMD 281014) in congenital learned helpless male rats in the forced swim test. The administration of EMD-281014 (0.3-30 mg/kg i.p.) to congenital learned helpless rats dose-dependently and significantly (at 10 and 30 mg/kg) decreased immobility and increased swimming compared to vehicle-treated animals. Thus, EMD 281014 produces effects in the forced swim test resembling those of antidepressants. PMID- 14755635 TI - Development of an integrative transformation system for the opportunistic pathogenic yeast Candida lusitaniae using URA3 as a selection marker. AB - The nucleotide sequence of the URA3 gene encoding orotidine-5'-phosphate decarboxylase (OMP DCase) of the human opportunistic pathogen yeast Candida lusitaniae was determined by degenerate PCR and chromosome walking. Deduced amino acid sequence showed strong homologies (59-85% identity) with OMP DCases of different Saccharomycetales and allowed identification of the known conserved domains. Very close upstream from the URA3 gene, the 3'-end of a gene encoding a Gea2-like protein was identified. A non-revertible C. lusitaniae ura3 mutant was selected on the basis of 5-fluoroorotic acid resistance. The mutation was a single point mutation resulting in the amino acid substitution D95V in a highly conserved domain, and in a concomitant EcoRV restriction site polymorphism. The mutant strain was successfully transformed to prototrophy following electroporation with the URA3 gene cloned in an integrative vector, with frequencies of 100-200 transformants per micro g of DNA. Southern blot analysis revealed that almost all transformants were derived from homologous recombination events at the resident locus. The GeneBank Accession No. for C. lusitaniae URA3 gene is AF450297. PMID- 14755636 TI - [URE3] prion propagation is abolished by a mutation of the primary cytosolic Hsp70 of budding yeast. AB - [URE3] and [PSI(+)] are infectious protein forms of the Saccharomyces cerevisiae Ure2p and Sup35p, respectively. We isolated an allele of SSA2, the primary cytosolic Hsp70, in a screen for mutants unable to maintain [URE3]. Designated ssa2-10, the mutation results in a leucine substitution for proline 395, a conserved residue of the peptide-binding domain. This allele also unexpectedly destabilizes [URE3] in newly formed heterozygotes: [URE3] is either absent in heterozygotes formed by crossing wild-type [URE3] cells with ssa2-10 mutants, or present and fully stable. SSA2 deletion mutants are weakly capable of maintaining [URE3]. The ssa2-10 allele is compatible with propagation of [PSI(+)]. However, in combination with a deletion of SSA1, ssa2-10 eliminates the nonsense suppression phenotype of [PSI(+)] cells. PMID- 14755637 TI - Isolation of a GPD gene from Debaryomyces hansenii encoding a glycerol 3 phosphate dehydrogenase (NAD+). AB - A gene homologous to GPD1, coding for glycerol-3-phosphate dehydrogenase (sn glycerol 3-phosphate: NAD(+) oxidoreductase, EC 1.1.1.8), has been isolated from the halophilic yeast Debaryomyces hansenii by complementation of a Saccharomyces cerevisiae gpd1 Delta mutant. DNA sequencing of the complementing genomic clone indicated the existence of an open reading frame encoding a protein with 369 amino acids. Comparative analysis of the deduced amino acid sequence showed high similarity to homologous genes described for other eukaryotic GPD enzymes. The sequence has been submitted to the GenBank database under Accession No. AY333427. PMID- 14755638 TI - Binding of Cdc48p to a ubiquitin-related UBX domain from novel yeast proteins involved in intracellular proteolysis and sporulation. AB - The Cdc48/p97 AAA-ATPase functions in membrane fusion and ubiquitin-dependent protein degradation. Here, we show that, in yeast, Cdc48p interacts with three novel proteins, Cuil-3p, which contain a conserved ubiquitin-related (UBX) domain. Cui2p and Cui3p are closely related, interact with each other, and are localized at the perinuclear membrane. Cdc48p binds directly the UBX domain of Cui3p in vitro. Multiple deletions of the CUI1, CUI2 and CUI3 genes confer deficiency in sporulation and degradation of model ubiquitin-protein fusions. The Cuil-3 proteins were also found to interact with Ufd3p, a WD repeat protein known to associate with Cdc48p. Together, these results indicate that the Cuil-3 proteins form complexes that are components of the ubiquitin-proteasome system. PMID- 14755639 TI - Endogenous NADPH-dependent aldose reductase activity influences product formation during xylose consumption in recombinant Saccharomyces cerevisiae. AB - Introduction of the xylose pathway from Pichia stipitis into Saccharomyces cerevisiae enables xylose utilization in recombinant S. cerevisiae. However, xylitol is a major by-product. An endogenous aldo-keto reductase, encoded by the GRE3 gene, was expressed at different levels in recombinant S. cerevisiae strains to investigate its effect on xylose utilization. In a recombinant S. cerevisiae strain producing only xylitol dehydrogenase (XDH) from P. stipitis and an extra copy of the endogenous xylulokinase (XK), ethanol formation from xylose was mediated by Gre3p, capable of reducing xylose to xylitol. When the GRE3 gene was overexpressed in this strain, the xylose consumption and ethanol formation increased by 29% and 116%, respectively. When the GRE3 gene was deleted in the recombinant xylose-fermenting S. cerevisiae strain TMB3001 (which possesses xylose reductase and XDH from P. stipitis, and an extra copy of endogenous XK), the xylitol yield decreased by 49% and the ethanol yield increased by 19% in anaerobic continuous culture with a glucose/xylose mixture. Biomass was reduced by 31% in strains where GRE3 was deleted, suggesting that fine-tuning of GRE3 expression is the preferred choice rather than deletion. PMID- 14755640 TI - Ion channel activity by Pichia membranifaciens killer toxin. AB - The cytocidal effect of Pichia membranifaciens killer toxin on Candida boidinii cells was studied. The halotolerant yeast P. membranifaciens CYC 1106 produces a unique 18 kDa killer toxin that exerts its killer activity against C. boidinii IGC 3430 only in the presence of NaCl. Metabolic events associated with the loss of C. boidinii IGC 3430 viability were quantitatively identical to those known to occur with K1 killer toxin-treated sensitive strains of Saccharomyces cerevisiae. The death of sensitive cells was characterized by a leakage of potassium, an influx of sodium and a decrease in intracellular pH. These effects occurred prior to and concomitantly with cell death, indicating that they were primary effects of the action of the toxin. Here we report that this protein forms ion-permeable channels in liposome membranes. These channels are freely permeable to common physiological ions. We suggest that channel formation is the cytotoxic mechanism of action of P. membranifaciens killer toxin. The channels described here are sufficiently non-selective to mediate cell death through a discharge of cellular membrane potential and changes in ionic homeostasis. No specific effects against killer toxin-treated sensitive cells were observed when the cell cycle was analysed. PMID- 14755641 TI - DsdA (D-serine deaminase): a new heterologous MX cassette for gene disruption and selection in Saccharomyces cerevisiae. AB - Dominant drug resistance markers offer experimental flexibility in the study of Saccharomyces cerevisiae by eliminating the dependence on auxotrophic mutations and, because they are phenotypically neutral, avoid the deleterious effects of auxotrophic mutations. We have developed a new dominant resistance marker, dsdAMX4, for use in the genetic manipulation of S. cerevisiae. The dsdA gene, which is derived from Escherichia coli and encodes a D-serine deaminase, confers to S. cerevisiae resistance to D-serine and the ability to use D-serine as a nitrogen source. Here we describe the construction of a dsdAMX4 cassette, capable of expression in S. cerevisiae, and the characterization of this new marker for use in chromosomal gene disruption. The unique selection properties of the dsdAMX4 cassette make it an important addition to the existing array of S. cerevisiae genetic tools. PMID- 14755642 TI - Monofunctional catalase P of Paracoccidioides brasiliensis: identification, characterization, molecular cloning and expression analysis. AB - Within the context of studies on genes from Paracoccidioides brasiliensis (Pb) potentially associated with fungus-host interaction, we isolated a 61 kDa protein, pI 6.2, that was reactive with sera of patients with paracoccidioidomycosis. This protein was identified as a peroxisomal catalase. A complete cDNA encoding this catalase was isolated from a Pb cDNA library and was designated PbcatP. The cDNA contained a 1509 bp ORF containing 502 amino acids, whose molecular mass was 57 kDa, with a pI of 6.5. The translated protein PbCATP revealed canonical motifs of monofunctional typical small subunit catalases and the peroxisome-PTS-1-targeting signal. The deduced and the native PbCATP demonstrated amino acid sequence homology to known monofunctional catalases and was most closely related to catalases from other fungi. The protein and mRNA were diminished in the mycelial saprobic phase compared to the yeast phase of infection. Protein synthesis and mRNA levels increased during the transition from mycelium to yeast. In addition, the catalase protein was induced when cells were exposed to hydrogen peroxide. The identification and characterization of the PbCATP and cloning and characterization of the cDNA are essential steps for investigating the role of catalase as a defence of P. brasiliensis against oxygen dependent killing mechanisms. These results suggest that this protein exerts an influence in the virulence of P. brasiliensis. PMID- 14755644 TI - 1H NMR detection of vitamin C in human brain in vivo. AB - Vitamin C (ascorbate) is well established as an essential nutrient that functions as an antioxidant. Since it is present in the human brain at detectable concentrations, this study was designed to detect and quantify ascorbate in the human brain in vivo using 1H NMR spectroscopy (MRS). Ascorbate was consistently detected in all five study subjects, and was measured using MEGA-PRESS difference editing. The in vivo resonance pattern was consistent with that of ascorbate based on position, line width, peak pattern, and relative intensity. Metabolites with a potential for coediting were assessed using phantom solutions. The putative resonances of myo-inositol, lactate, glycerophosphocholine, phosphocholine, and phosphoethanolamine were detected at positions distinct from those of ascorbate. This study represents the first in vivo detection of vitamin C in the human brain using 1H MRS. A concentration of 1.3 +/- 0.3 micromol/g (mean +/- SD, N = 4) was estimated. PMID- 14755645 TI - SENSE-DTI at 3 T. AB - While holding vast potential, diffusion tensor imaging (DTI) with single excitation protocols still faces serious challenges. Limited spatial resolution, susceptibility to magnetic field inhomogeneity, and low signal-to-noise ratio (SNR) may be considered the most prominent limitations. It is demonstrated that all of these shortcomings can be effectively mitigated by the transition to parallel imaging technology and high magnetic field strength. Using the sensitivity encoding (SENSE) technique at 3 T, brain DTI was performed in nine healthy volunteers. Despite enhanced field inhomogeneity, parallel acquisition permitted both controlling geometric distortions and enhancing spatial resolution up to 0.8 mm in-plane. Heightened SNR requirements were met in part by high base sensitivity at 3 T. A further significant increase in SNR efficiency was accomplished by SENSE acquisition, exploiting enhanced encoding speed for echo time reduction. Based on the resulting image data, high-resolution tensor mapping is demonstrated. PMID- 14755646 TI - Spiral MR myocardial tagging. AB - In the present study, complementary spatial modulation of magnetization (CSPAMM) myocardial tagging was extended with an interleaved spiral imaging sequence. The use of a spiral sequence enables the acquisition of grid-tagged images with a tagline distance as low as 4 mm in a single breath-hold. Alternatively, a high temporal resolution of 77 frames per second was obtained with 8-mm grid spacing. Ten healthy adult subjects were studied. With this new approach, high-quality images can be obtained and the tags persist throughout the entire cardiac cycle. PMID- 14755647 TI - Fat and water separation in balanced steady-state free precession using the Dixon method. AB - In this work the feasibility of separating fat and water signals using the balanced steady-state free precession (SSFP) technique is demonstrated. The technique is based on the observation (Scheffler and Hennig, Magnetic Resonance in Medicine 2003;49:395-397) that at the nominal values of TE = TR/2 in SSFP imaging, phase coherence can be achieved at essentially only two orientations (0 degrees and 180 degrees ) relative to the RF pulses in the rotating frame, under the assumption of TR << T2, and independently of the SSFP angle. This property allows in-phase and out-of-phase SSFP images to be obtained by proper choices of the center frequency offset, and thus allows the Dixon subtraction method to be utilized for effective fat-water separation. The TR and frequency offset for optimal fat-water separation are derived from theories. Experimental results from healthy subjects, using a 3.0 Tesla system, show that nearly complete fat suppression can be accomplished. PMID- 14755648 TI - MRI detection of early endothelial activation in brain inflammation. AB - MRI is an increasingly important clinical tool, but it is clear that conventional imaging fails to identify the full extent of lesion load in certain conditions, such as multiple sclerosis. The aim of this study was to determine whether a novel contrast agent (Gd-DTPA-B(sLeX)A, which contains an sLeX mimetic moiety that enables it to bind to the adhesion molecule E-selectin) can be used to identify endothelial activation in the brain. Microinjection of the proinflammatory cytokines IL-1beta or TNF-alpha into the striatum of Wistar rats rapidly induces focal adhesion molecule expression on the endothelium in the absence of MRI-visible changes. This phenomenon was used to investigate the potential of Gd-DTPA-B(sLeX)A to reveal MRI-invisible brain pathology. T1 weighted serial images were acquired in anesthetized animals before and after administration of Gd-DTPA-B(sLeX)A, 3-4 hr after cytokine was injected intracerebrally. Both TNF-alpha and IL-1beta up-regulated E-selectin on the brain endothelium, which correlated with increased signal intensity observed after administration of the novel contrast agent. No enhancement was visible with the nonselective contrast agent Gd-DTPA-BMA, indicating that there was no leakage of the agent across the blood-brain barrier (BBB) or nonselective binding to the endothelium. These data demonstrate the potential of such contrast agents for the early detection of brain injury and inflammation. PMID- 14755649 TI - Distribution of intramyocellular lipids in human calf muscles as determined by MR spectroscopic imaging. AB - In this study the distribution of intramyocellular lipids (IMCL) in human calf muscles was determined by 1H-MR spectroscopic imaging (MRSI) measurements. An obstacle for MRSI measurements in the calf, including different muscles, is the inevitable inclusion of regions with high concentrations of extramyocellular lipids (EMCL). This can lead to signal bleeding and consequently to unpredictable overlaps of IMCL resonances with EMCL in voxels of interest. The results of this study show that signal bleeding from EMCL can be substantially reduced in voxels from calf muscles by the application of a lipid extrapolation (LE) procedure (Haupt et al., Magn Reson Med 1996;35:678). The spectra of all voxels located within muscle tissue were fitted, and the metabolite values were assigned to one of 10 different muscles based on image segmentation. Significant IMCL differences between some muscles were obtained, with high values in m. soleus and two to three times lower values in the tibialis anterior, tibialis posterior, and gastrocnemius muscles. In addition to gross differences between muscles, significant intersubject differences were observed in both IMCL content and distribution over different muscles. A significant correlation between fiber orientation (obtained from orientation-dependent dipolar coupling of creatine and taurine resonances) and IMCL content was found, indicating that IMCL content is directly correlated to biomechanical properties. PMID- 14755650 TI - Strategy for the spectral filtering of myo-inositol and other strongly coupled spins. AB - A multiple quantum filter strategy is presented for spectrally discriminating metabolites with strongly coupled spins from those whose spins are either uncoupled or weakly coupled. The strategy also includes a means for selectively suppressing the background multiplets of metabolites that also have strongly coupled spins. As a demonstration of its efficacy at 3.0 T, the strategy is shown to enhance by a factor of approximately 5 the signal-to-background ratio of the myo-inositol band at 3.6 ppm relative to that in response to a PRESS sequence with the same sequence timings. This is done by eliminating the uncoupled resonance of glycine and the weakly coupled multiplets of glutamate and glutamine, and by selectively suppressing the strongly coupled taurine multiplet 3-fold. The macromolecular background was effectively removed through its transverse decay over 105 ms. The associated cost of gaining the signal to background enhancement is a drop in signal yield by a factor of 0.75 relative to PRESS at the same timings. The myo-inositol signal to noise ratio was nevertheless maintained by the filter at approximately 12. PMID- 14755651 TI - Noninvasive monitoring of stem cell transfer for muscle disorders. AB - In this study the ability of magnetodendrimers to efficiently label cultured muscle stem cells and allow for subsequent in vivo cell detection was determined. Magnetodendrimer-labeled cells exhibited normal growth rates in culture, and retained their capacity to undergo proliferation and form normal myotubes. Labeled stem cells possessed high in vivo proton relaxivities that enhanced MRI contrast properties and enabled us to noninvasively monitor the stem cells' incorporation into dystrophic muscle. Well defined regions of decreased signal intensity were observed in both T2- and T1-weighted image sequences. MRI was used to longitudinally follow stem cell dynamics in dystrophic muscle with in-plane resolutions on the order of a single muscle fiber (22 x 43 microm2). Regions of decreased signal intensity were well correlated with iron accumulation and other histochemical markers of stem cell incorporation. We concluded that this technique may be useful for continuous noninvasive readouts of stem cell transfer, replacing sequential muscle biopsies and tissue harvesting. PMID- 14755653 TI - dGEMRIC (delayed gadolinium-enhanced MRI of cartilage) indicates adaptive capacity of human knee cartilage. AB - Delayed gadolinium-enhanced MRI of cartilage (dGEMRIC) is a new imaging technique to estimate joint cartilage glycosaminoglycan content by T1-relaxation time measurements after penetration of the hydrophilic contrast agent Gd-DTPA(2-). This study compares dGEMRIC in age-matched healthy volunteers with different levels of physical activity: Group 1 (n = 12): nonexercising individuals; Group 2 (n = 16): individuals with physical exercise averaging twice weekly; Group 3 (n = 9): male elite runners. dGEMRIC was performed 2 hr after an intravenous injection of Gd-DTPA(2-) at 0.3 mmol/kg body weight. T1 differed significantly between the three different levels of physical exercise. T1 values (mean of medial and lateral femoral cartilage) for Groups 1, 2, and 3 were: 382 +/- 33, 424 +/- 22 and 476 +/- 36, respectively (ms, mean +/- SD) (P = 0.0004, 1 vs. 2 and 0.0002, 2 vs. 3). Irrespective of the exercise level, T1 was longer in lateral compared to medial femoral cartilage (P = 0.00005; n = 37). In conclusion, this cross sectional study indicates that human knee cartilage adapts to exercise by increasing the glycosaminoglycan content. Furthermore, results suggest a compartmental difference within the knee with a higher glycosaminoglycan content in lateral compared to medial femoral cartilage. A higher proportion of extracellular water, i.e., larger distribution volume, may to some extent explain the high T1 in the elite runners. PMID- 14755652 TI - The existence of biexponential signal decay in magnetic resonance diffusion weighted imaging appears to be independent of compartmentalization. AB - It is generally believed that the apparent diffusion coefficient (ADC) changes measured by diffusion-weighted imaging (DWI) in brain pathologies are related to alterations in the water compartments. The aim of this study was to elucidate the role of compartmentalization in DWI via biexponential analysis of the signal decay due to diffusion. DWI experiments were performed on mouse brain over an extended range of b-values (up to 10,000 mm(-2) s) under intact, global ischemic, and cold-injury conditions. DWI was additionally applied to centrifuged human erythrocyte samples with a negligible extracellular space. Biexponential signal decay was found to occur in the cortex of the intact mouse brain. During global ischemia, in addition to a drop in the ADC in both components, a shift from the volume fraction of the rapidly diffusing component to the slowly diffusing one was observed. In cold injury, the biexponential signal decay was still present despite the electron-microscopically validated disintegration of the membranes. The biexponential function was also applicable for fitting of the data obtained on erythrocyte samples. The results suggest that compartmentalization is not an essential feature of biexponential decay in diffusion experiments. PMID- 14755654 TI - Detection of simulated pulmonary embolism in a porcine model using hyperpolarized 3He MRI. AB - Several radiological imaging modalities are available to assist with the clinical diagnosis of pulmonary embolism (PE). The most frequently used techniques-nuclear medicine ventilation-perfusion (VP) scan, computed tomography (CT), magnetic resonance angiography (MRA), and pulmonary angiography (PA)-all have literature supported, substantial limitations with respect to timeliness and patient safety. Hyperpolarized 3He magnetic resonance gas distribution imaging (HP 3He MRI) recently has shown potential as a safer and faster alternative. In this study, we performed HP 3He MRI on a porcine model (N = 6) of simulated PE using selective occlusion balloon catheterization (N = 4) and nonselective aged autologous clot injection (N = 1). The technique was also performed on a normal pig and again after the animal was killed. Temporal depletion of regional HP 3He MRI signal intensity provided for a qualitative assessment of simulated PE (N = 4), and regional PAO2 (alveolar partial pressure of oxygen) was calculated in affected airspaces for a quantitative assessment of simulated PE (N = 1). The preliminary results suggest that HP (3)He MRI shows promise as a means of assessing regional pulmonary perfusion abnormalities in the porcine models of simulated PE that were used in this study. PMID- 14755655 TI - Regional variations in normal brain shown by quantitative magnetization transfer imaging. AB - A quantitative magnetization transfer imaging (qMTI) study, based on a two-pool model of magnetization transfer, was performed on seven normal subjects to determine, on a regional basis, normal values for the pool sizes, exchange, and relaxation parameters that characterize the MT phenomenon. Regions were identified on high-resolution anatomical scans using a combination of manual and automatic methods. Only voxels identified as pure tissue at the resolution of the quantitative scans were considered for analysis. While no left/right differences were observed, significant differences were found among white-matter regions and gray-matter regions. These regional differences were compared with existing cytoarchitectural data. In addition, the pattern and magnitude of the regional differences observed in white matter was found to be different from that reported previously for an alternative putative MRI measure of myelination, the 10-50-ms T2 component described as myelin water. PMID- 14755656 TI - Longitudinal MRI tracking of the angiogenic response to hind limb ischemic injury in the mouse. AB - Ischemic injury and revascularization are frequently associated with hyperpermeability. Although extravasation of plasma proteins may promote tissue recovery through the generation of the provisional matrix that supports angiogenesis, edema may also result in progressive damage to the muscle. The aim of this research was to determine the time course of hyperpermeability associated with the angiogenic response induced by ligation of the femoral artery at the right posterior limb in mice. Hyperpermeability was followed noninvasively by MRI using an in-house-built permanent polyethylene catheter that enabled daily intravenous administration of biotin-BSA-Gd-DTPA. The mice were scanned once prior to ligation and five times during the week post-ligation. The MRI data, along with histopathology, indicated that the early hemodynamic compensation over loss of arterial blood supply occurred by angiogenesis and dilation of vessels in the skin and subcutaneous fat, and was accompanied by vascular hyperpermeability around the site of ligation. Functional recovery of the ischemic limb (i.e., regaining the ability to step on the limb), and the color and shape of the toes correlated with regeneration as shown by histopathology and MRI analysis. Thus, MRI provided valuable information on the transient hyperpermeability induced during the early stages of angiogenesis, and its subsequent resolution along with functional recovery from acute hind limb ischemia in mice. PMID- 14755657 TI - Comparison of FAIR perfusion kinetics with DSC-MRI and functional histology in a model of transient ischemia. AB - Flow-sensitive alternating inversion recovery (FAIR) is a noninvasive method for perfusion imaging. It has been shown that the FAIR signal may depend on hemodynamic parameters other than perfusion, the most important one being transit delays of labeled spins to the observed tissue. These parameters are expected to change with ischemia. The goal of this study was to assess the effect of these changes on the interpretation of FAIR results in the case of altered perfusion. This was investigated in a rat model of transient cerebral ischemia. It was shown that the ratio of FAIR signal in the infarct compared to the contralateral side was lower at short inflow times, which suggests that transit times affected the effective FAIR signal. The FAIR results were compared with those from functional histology and dynamic susceptibility contrast MRI, and the findings indicated that the altered kinetics of the FAIR signal were related to reduced and delayed inflow in the infarct region--not to a decrease in the number of functional vessels. PMID- 14755658 TI - Biexponential diffusion tensor analysis of human brain diffusion data. AB - Several studies have shown that in tissues over an extended range of b-factors, the signal decay deviates significantly from the basic monoexponential model. The true nature of this departure has to date not been identified. For the current study, line scan diffusion images of brain suitable for biexponential diffusion tensor analysis were acquired in normal subjects on a clinical MR system. For each of six noncollinear directions, 32 images with b-factors ranging from 5 to 5000 s/mm2 were collected. Biexponential fits yielded parameter maps for a fast and a slow diffusion component. A subset of the diffusion data, consisting of the images obtained at the conventional range of b-factors between 5 and 972 s/mm2, was used for monoexponential diffusion tensor analysis. Fractional anisotropy (FA) of the fast-diffusion component and the monoexponential fit exhibited no significant difference. FA of the slow-diffusion biexponential component was significantly higher, particularly in areas of lower fiber density. The principal diffusion directions for the two biexponential components and the monoexponential solution were largely the same and in agreement with known fiber tracts. The second and third diffusion eigenvector directions also appeared to be aligned, but they exhibited significant deviations in localized areas. PMID- 14755659 TI - Noquist: reduced field-of-view imaging by direct Fourier inversion. AB - A novel technique called "Noquist" is introduced for the acceleration of dynamic cardiac magnetic resonance imaging (CMRI). With the use of this technique, a more sparsely sampled dynamic image sequence is reconstructed correctly, without Nyquist foldover artifact. Unlike most other reduced field-of-view (rFOV) methods, Noquist does not rely on data substitution or temporal interpolation to reconstruct the dynamic image sequence. The proposed method reduces acquisition time in dynamic MRI scans by eliminating the data redundancy associated with static regions in the dynamic scene. A reduction of imaging time is achieved by a fraction asymptotically equal to the static fraction of the FOV, by omitting acquisition of an appropriate subset of phase-encoding views from a conventional equidistant Cartesian acquisition grid. The theory behind this method is presented along with sample reconstructions from real and simulated data. Noquist is compared with conventional cine imaging by retrospective selection of a reduced data set from a full-grid conventional image sequence. In addition, a comparison is presented, using real and simulated data, of our technique with an existing rFOV technique that uses temporal interpolation. The experimental results confirm the theory, and demonstrate that Noquist reduces scan time for cine MRI while fully preserving both spatial and temporal resolution, but at the cost of a reduced signal-to-noise ratio (SNR). PMID- 14755660 TI - Iterative Next-Neighbor Regridding (INNG): improved reconstruction from nonuniformly sampled k-space data using rescaled matrices. AB - The reconstruction of MR images from nonrectilinearly sampled data is complicated by the fact that the inverse 2D Fourier transform (FT) cannot be performed directly on the acquired k-space data set. k-Space gridding is commonly used because it is an efficient reconstruction method. However, conventional gridding requires optimized density compensation functions (DCFs) to avoid profile distortions. Oftentimes, the calculation of optimized DCFs presents an additional challenge in obtaining an accurately gridded reconstruction. Another type of gridding algorithm, the block uniform resampling (BURS) algorithm, often requires singular value decomposition (SVD) regularization to avoid amplification of data imperfections, and under some conditions it is difficult to adjust the regularization parameters. In this work, new reconstruction algorithms for nonuniformly sampled k-space data are presented. In the newly proposed algorithms, high-quality reconstructed images are obtained from an iterative reconstruction that is performed using matrices scaled to sizes greater than that of the target image matrix. A second version partitions the sampled k-space region into several blocks to avoid limitations that could result from performing multiple 2D-FFTs on large data matrices. The newly proposed algorithms are a simple alternative approach to previously proposed optimized gridding algorithms. PMID- 14755661 TI - FAIR true-FISP perfusion imaging of the kidneys. AB - Most arterial spin labeling (ASL) techniques apply echoplanar imaging (EPI) because this strategy provides relatively high SNR in short measuring times. Unfortunately, those techniques are very susceptible to static magnetic field inhomogeneities and perfusion signals from organs with fast transverse relaxation might decrease due to the exchange of water molecules in capillaries and organ tissue combined with relatively long echo times of EPI sequences. To overcome these problems a novel imaging technique, FAIR True-FISP, was developed. It combines a FAIR (flow-sensitive alternating inversion recovery) perfusion preparation and a true fast imaging with steady precession (True-FISP) data acquisition strategy. True-FISP was chosen since this sequence type does not show the mentioned disadvantages of EPI, but provides a similar SNR per measuring time. An important problem of this approach is that True-FISP sequences usually work in a steady state which is independent of a previous preparation of magnetization. For this reason a sequence structure had to be developed which keeps the advantages of True-FISP and makes the signal intensity sensitive to the FAIR preparation. Breathhold and nonbreathhold examinations of kidneys are presented and possible strategies to quantitative flow measurements are reported. It is shown that correction of spatially inhomogeneous receiver coil characteristics is easily feasible and leads to clinically valuable perfusion examinations of kidneys without application of potentially nephrotoxic contrast media. PMID- 14755662 TI - Pulse sequence for multislice T1rho-weighted MRI. AB - A 2D multislice spin-lock (MS-SL) MR pulse sequence is presented for rapid volumetric T1rho-weighted imaging. Image quality is compared with T1rho-weighted data collected using a single-slice (SS) SL sequence and T2-weighted data from a standard MS spin-echo (SE) sequence. Saturation of longitudinal magnetization by the application of nonselective SL pulses is experimentally measured and theoretically modeled as T2rho decay. The saturation data is used to correct the image data as a function of the SL pulse duration to make quantitative measurements of T1rho. Measurements of T1rho using the saturation-corrected MS-SL data are nearly identical to those measured using an SS-SL sequence. The MS-SL sequence produces quantitative T1rho maps of an entire sample volume with the high-SNR advantages conferred by SE-based sequences. PMID- 14755663 TI - Quantification of multicontrast vascular MR images with NLSnake, an active contour model: in vitro validation and in vivo evaluation. AB - Vessel-wall measurements from multicontrast MRI provide information on plaque structure and evolution. This requires the extraction of numerous contours. In this work a contour-extraction method is proposed that uses an active contour model (NLSnake) adapted for a wide range of MR vascular images. This new method employs length normalization for the purpose of deformation computation and offers the advantages of simplified parameter tuning, fast convergence, and minimal user interaction. The model can be initialized far from the boundaries of the region to be segmented, even by only one pixel. The accuracy and reproducibility of NLSnake endoluminal contours were assessed on vascular phantom MR angiography (MRA) and high-resolution in vitro MR images of rabbit aorta. An in vivo evaluation was performed on rabbit and clinical data for both internal and external vessel-wall contours. In phantoms with 95% stenoses, NLSnake measured 94.3% +/- 3.8%, and the accuracy was even better for milder stenoses. In the images of rabbit aorta, variability between NLSnake and experts was less than interobserver variability, while the maximum intravariability of NLSnake was equal to 1.25%. In conclusion, the NLSnake technique successfully quantified the vessel lumen in multicontrast MR images using constant parameters. PMID- 14755664 TI - Active deep brain stimulation during MRI: a feasibility study. AB - The goal of this study was to evaluate the feasibility of active deep brain stimulation (DBS) during the application of standard clinical sequences for functional MRI (fMRI) in phantom measurements. During active DBS, we investigated induced voltage, temperature at the electrode tips and lead, forces on the electrode and lead, consequences of defective leads and loose connections, proper operation of the neurostimulator, and image quality. Sequences for diffusion- and perfusion-weighted imaging, fMRI, and morphologic MRI were used. The DBS electrode and lead were placed in a NaCl solution-filled phantom. The results indicate that there are severe potential hazards for patients. Strong heating, high induced voltage, and even sparking at defects in the connecting cable could be observed. However, it was demonstrated that under certain conditions, safe MR examinations during active DBS are feasible. Certain safety precautions are recommended in this report. PMID- 14755665 TI - Influence of body temperature on the BOLD effect in murine SCC tumors. AB - Changes in the blood oxygen level dependent (BOLD) enhancements in tumors (squamous cell carcinoma, (SCCVII)) implanted in mice maintained at core temperatures of 30 degrees C or 37 degrees C were measured using MRI and compared to tumor oxygen levels obtained using an oxygen-sensitive Eppendorf electrode. Tumors were implanted in a hindleg of the mice intramuscularly. Tumor-bearing mice were imaged by BOLD MRI, while first breathing air and then carbogen (95% O2, 5% CO2) for 15-min intervals at a core temperature of 30 degrees C. After an equilibration period, the identical regimen was conducted with the same animal maintained at 37 degrees C. This procedure was repeated with additional mice starting at 37 degrees C followed by imaging at 30 degrees C. Likewise, oxygen electrode measurements of the tumor were determined at core temperatures of 30 degrees C and 37 degrees C. The Eppendorf measurements showed that tumors in animals maintained at 30 degrees C were significantly more hypoxic than at 37 degrees C. MRI studies demonstrated stronger BOLD enhancement at 30 degrees C than at 37 degrees C, suggesting significant changes in hypoxia and/or blood flow in tumors at these temperatures. The findings of the study stress the importance of maintaining normal core temperature when assessing tumor oxygen status using functional imaging modalities or oxygen-sensitive electrodes. PMID- 14755666 TI - Steady-state diffusion-weighted imaging of in vivo knee cartilage. AB - Diffusion-weighted imaging (DWI) has strong potential as a diagnostic for early cartilage damage, with clinical impact for diseases such as osteoarthritis. However, in vivo DWI of cartilage has proven difficult with conventional methods due to the short T2. This work presents a 3D steady-state DWI sequence that is able to image short-T2 species with high SNR. When combined with 2D navigator correction of motion-induced phase artifacts, this method enables high resolution in vivo DWI of cartilage. In vivo knee images in healthy subjects are presented with high SNR (SNR = 110) and submillimeter in-plane resolution (0.5 x 0.7 x 3.0 mm(3)). A method for fitting the diffusion coefficient is presented which produces fits within 10% of literature values. This method should be applicable to other short-T2 tissues, such as muscle, which are difficult to image using traditional DWI methods. PMID- 14755667 TI - Event-related fMRI technique for auditory processing with hemodynamics unrelated to acoustic gradient noise. AB - Acoustic noise from the imaging gradients presents a major difficulty in functional MRI (fMRI) studies of auditory cortical function. For studies involving hearing-impaired pediatric subjects, the auditory stimuli should be presented during completely silent gradient intervals. In addition, the scan time is limited by constraints involving subject motion and subject compliance. A novel event-related method for conducting fMRI studies of auditory function is proposed. Auditory stimuli are presented during completely silent gradient intervals, but using a variable TR. A general nonlinear model (GNLM) is proposed as a postprocessing methodology for the data. The technique increases the flexibility of the experimental design, with minimal loss of sensitivity compared to standard fMRI acquisition techniques, and may therefore be useful for fMRI studies of auditory function in hearing-impaired pediatric subjects. PMID- 14755668 TI - Floating navigator echo (FNAV) for in-plane 2D translational motion estimation. AB - A modification of the classical navigator echo (NAV) technique is presented whereby both 2D translational motion components are computed from a single navigator line. Instead of acquiring the NAV at the center of the k-space, a kx line is acquired off-center in the phase-encoding (ky) direction as a floating NAV (FNAV). It is shown that the translational motion in both the readout and phase-encoding directions can be computed from this line. The algorithm used is described in detail and verified experimentally. The new technique can be readily implemented to replace classic NAV in MRI sequences, with little to no additional cost or complexity. The new method can help suppress 2D translational motion and provide more accurate motion estimates for other motion-suppression techniques, such as the diminishing variance algorithm. PMID- 14755669 TI - Artifact suppression in imaging of myocardial infarction using B1-weighted phased array combined phase-sensitive inversion recovery. AB - Regions of the body with long T1, such as cerebrospinal fluid (CSF), may create ghost artifacts on gadolinium-hyperenhanced images of myocardial infarction when inversion recovery (IR) sequences are used with a segmented acquisition. Oscillations in the transient approach to steady state for regions with long T1 may cause ghosts, with the number of ghosts being equal to the number of segments. B1-weighted phased-array combining provides an inherent degree of ghost artifact suppression because the ghost artifact is weighted less than the desired signal intensity by the coil sensitivity profiles. Example images are shown that illustrate the suppression of CSF ghost artifacts by the use of B1-weighted phased-array combining of multiple receiver coils. PMID- 14755670 TI - Does fractional anisotropy have better noise immunity characteristics than relative anisotropy in diffusion tensor MRI? An analytical approach. AB - Fractional anisotropy (FA) and relative anisotropy (RA) are the two most commonly used scalar measures of anisotropy in diffusion tensor (DT) MRI. While a few published studies have shown that FA has superior noise immunity relative to RA, no theoretical basis has been proposed to explain this behavior. In the current study, the diffusion tensor invariants were used to derive a simple analytical expression that directly relates RA and FA. An analysis based on that analytical expression demonstrated that the FA images have a higher signal-to-noise ratio (SNR) than RA for any value of tensor anisotropy RA or FA > 0. This theoretical behavior was verified using both Monte Carlo simulations and bootstrap analysis of DT-MRI data acquired in a spherical water phantom and normal human subjects. PMID- 14755671 TI - Comparison of Fourier and wavelet resampling methods. AB - Resampling can be used to compute the null distribution of any test statistic for the purpose of measuring the significance of the measured value. This study investigated how well the spatial and temporal correlations of simulated and experimentally observed fMRI time series were preserved under Fourier and wavelet resampling methods. The null distributions of a test statistic estimated by each resampling method were compared. In addition, both resampling methods were applied to locate activated voxels in an fMRI dataset and ROC analysis showed that wavelet resampling performed more accurately than Fourier resampling. PMID- 14755672 TI - Enhancing measured diffusion anisotropy in gray matter by eliminating CSF contamination with FLAIR. AB - In this work, the effect of fluid-attenuated inversion recovery (FLAIR) on measured diffusion anisotropy was investigated in gray matter. DTI data were obtained with and without FLAIR in six normal volunteers. The application of FLAIR was experimentally demonstrated to lead to a consistent increase in fractional anisotropy (FA) in gray-matter regions, which was attributed to suppressed partial volume effects from CSF. In addition to these experimental results, Monte Carlo simulations were performed to ascertain the effect of noise on the measured FA under the experimental conditions of this study. The experimentally observed effect of noise was corroborated by the simulation, indicating that the increase in the measured FA was not due to a noise-related bias but to an actual increase in diffusion anisotropy. This enhanced measurement of diffusion anisotropy can be potentially used to differentiate directionally dependent structure and tracking fibers in gray matter. PMID- 14755673 TI - Quantitative diffusion imaging with steady-state free precession. AB - The addition of a single, unbalanced diffusion gradient to the steady-state free precession (SSFP) imaging sequence sensitizes the resulting signal to free diffusion. Unfortunately, the confounding influence of both longitudinal (T1) and transverse (T2) relaxation on the diffusion-weighted SSFP (dwSSFP) signal has made it difficult to quantitatively determine the apparent diffusion coefficient (ADC). Here, a multistep method in which the T1, T2, and spin density (Mo) constants are first determined using a rapid mapping technique described previously is presented. Quantitative ADC can then be determined through a novel inversion of the appropriate signal model. The accuracy and precision of our proposed method (which we term DESPOD) was determined by comparing resulting ADC values from phantoms to those calculated from traditional diffusion-weighted echo planar imaging (dwEPI) images. Error within the DESPOD-derived ADC maps was found to be less than 3%, with good precision over a biologically relevant range of ADC values. PMID- 14755675 TI - Prostate carcinoma: opportunities for translational research. AB - Adenocarcinoma of the prostate continues to be a major health concern. Although modern screening techniques have increased the number of men presenting with early stage disease, a significant population of men will present with intermediate or advanced pathological risk factors for recurrence. There are defined limitations in outcome with traditional therapies including surgery, radiation therapy, and hormone manipulation. Patients with intermediate and high risk factors for treatment failure are candidates for protocols using translational research strategies incorporated into studies currently in development. These strategies may be able to selectively treat expression products of tumor and thus be more selective in the target for treatment. Carefully designed studies using these translational strategies have great potential in improving clinical outcome, tumor kill, and normal tissue tolerance in the care of these patients. PMID- 14755676 TI - Chemoprevention of prostate cancer with selenium: an update on current clinical trials and preclinical findings. AB - Prostate cancer is the most common cancer diagnosed and the second leading cause of cancer-related deaths in men in the United States. The etiological factors that give rise to prostate cancer are not known. Therefore, it is not possible to develop primary intervention strategies to remove the causative agents from the environment. However, secondary intervention strategies with selenium (Se) compounds and other agents represent a viable option to reduce the morbidity and mortality of prostate cancer. In this review, we discuss ongoing clinical trials. In addition, we discuss preclinical mechanistic studies that provide insights into the biochemical and molecular basis for the anti-carcinogenic activity of both inorganic and organic forms of Se. PMID- 14755677 TI - Pathological and molecular mechanisms of prostate carcinogenesis: implications for diagnosis, detection, prevention, and treatment. AB - Prostate cancer is an increasing threat throughout the world. As a result of a demographic shift in population, the number of men at risk for developing prostate cancer is growing rapidly. For 2002, an estimated 189,000 prostate cancer cases were diagnosed in the U.S., accompanied by an estimated 30,200 prostate cancer deaths [Jemal et al., 2002]. Most prostate cancer is now diagnosed in men who were biopsied as a result of an elevated serum PSA (>4 ng/ml) level detected following routine screening. Autopsy studies [Breslow et al., 1977; Yatani et al., 1982; Sakr et al., 1993], and the recent results of the Prostate Cancer Prevention Trial (PCPT) [Thompson et al., 2003], a large scale clinical trial where all men entered the trial without an elevated PSA (<3 ng/ml) were subsequently biopsied, indicate the prevalence of histologic prostate cancer is much higher than anticipated by PSA screening. Environmental factors, such as diet and lifestyle, have long been recognized contributors to the development of prostate cancer. Recent studies of the molecular alterations in prostate cancer cells have begun to provide clues as to how prostate cancer may arise and progress. For example, while inflammation in the prostate has been suggested previously as a contributor to prostate cancer development [Gardner and Bennett, 1992; Platz, 1998; De Marzo et al., 1999; Nelson et al., 2003], research regarding the genetic and pathological aspects of prostate inflammation has only recently begun to receive attention. Here, we review the subject of inflammation and prostate cancer as part of a "chronic epithelial injury" hypothesis of prostate carcinogenesis, and the somatic genome and phenotypic changes characteristic of prostate cancer cells. We also present the implications of these changes for prostate cancer diagnosis, detection, prevention, and treatment. PMID- 14755678 TI - Effect of finasteride on risk of prostate cancer: how little we really know. AB - The Prostate Cancer Prevention Trial (PCPT) reported conclusively that finasteride prevents or delays the detection of prostate cancer. One perplexing finding was that more high-grade tumors were detected in the finasteride treated group. It is hard to put this into perspective because of the limited published data on the effects of finasteride on prostate cancer. The strong possibility exists that the increase in high-grade tumors may be due to a treatment effect, which causes intermediate grade cancers to appear to be high-grade or aggressive tumors. Confirmation of a spurious tumor grade "inflation" will make the conclusions of this study clearer and define the benefits of finasteride chemoprevention in a more favorable light. PMID- 14755679 TI - Androgen receptor: a key molecule in the progression of prostate cancer to hormone independence. AB - Despite earlier detection and recent advances in surgery and radiation, prostate cancer is second only to lung cancer in male cancer deaths in the United States. Hormone therapy in the form of medical or surgical castration remains the mainstay of systemic treatment in prostate cancer. Over the last 15 years with the clinical use of prostate specific antigen (PSA), there has been a shift to using hormone therapy earlier in the disease course and for longer duration. Despite initial favorable response to hormone therapy, over a period of time these tumors will develop androgen-independence that results in death. The androgen receptor (AR) is central to the initiation and growth of prostate cancer and to its response to hormone therapy. Analyses have shown that AR continues to be expressed in androgen-independent tumors and AR signaling remains intact as demonstrated by the expression of the AR regulated gene, PSA. Androgen independent prostate cancers have demonstrated a variety of AR alterations that are either not found in hormone naive tumors or found at lower frequency. These changes include AR amplification, AR point mutation, and changes in expression of AR co-regulatory proteins. These AR changes result in a "super AR" that can respond to lower concentrations of androgens or to a wider variety of agonistic ligands. There is also mounting evidence that AR can be activated in a ligand independent fashion by compounds such as growth factors or cytokines working independently or in combination. These growth factors working through receptor tyrosine kinase pathways may promote AR activation and growth in low androgen environments. The clinical significance of these AR alterations in the development and progression of androgen-independent prostate cancer remains to be determined. Understanding the changes in AR signaling in the evolution of androgen-independent prostate cancer will be key to the development of more effective hormone therapy. PMID- 14755680 TI - Estrogens and anti-estrogens: key mediators of prostate carcinogenesis and new therapeutic candidates. AB - Despite the historical use of estrogens in the treatment of prostate cancer (PCa) little is known about their direct biological effects on the prostate, their role in carcinogenesis, and what mechanisms mediate their therapeutic effects on PCa. It is now known that estrogens alone, or in synergism with an androgen, are potent inducers of aberrant growth and neoplastic transformation in the prostate. The mechanisms of estrogen carcinogenicity could be mediated via induction of unscheduled cell proliferation or through metabolic activation of estrogens to genotoxic metabolites. Age-related changes and race-/ethnic-based differences in circulating or locally formed estrogens may explain differential PCa risk among different populations. Loss of expression of estrogen receptor (ER)-beta expression during prostate carcinogenesis and prevention of estrogen-mediated oxidative damage could be exploited in future PCa prevention strategies. Re expression of ER-beta in metastatic PCa cells raises the possibility of using ER beta-specific ligands in triggering cell death in these malignant cells. A variety of new estrogenic/anti-estrogenic/selective estrogen receptor modulator (SERM)-like compounds, including 2-methoxyestradiol, genistein, resveratrol, licochalcone, Raloxifene, ICI 182,780, and estramustine are being evaluated for their potential in the next generation of PCa therapies. Increasing numbers of patients self-medicate with herbal formulations such as PC-SPES. Some of these compounds are selective ER-beta ligands, while most of them have minimal interaction with ER-alpha. Although many may inhibit testosterone production by blockade of the hypothalamal-pituitary-testis axis, the most effective agents also exhibit direct cytostatic, cytotoxic, or apoptotic action on PCa cells. Some of them are potent in interfering with tubulin polymerization, blocking angiogenesis and cell motility, suppressing DNA synthesis, and inhibiting specific kinase activities. Further discovery of other compounds with potent apoptotic activities but minimal estrogen action should promote development of a new generation of effective PCa preventive or treatment regimens with few or no side-effects due to estrogenicity. Further advancement of our knowledge of the role of estrogens in prostate carcinogenesis through metabolic activation of estrogens and/or ER-mediated pathways will certainly result in better preventive or therapeutic modalities for PCa. PMID- 14755681 TI - Par-4 inducible apoptosis in prostate cancer cells. AB - Prostate cancer is associated with the inability of prostatic epithelial cells to undergo apoptosis rather than with increased cell proliferation. Prostate apoptosis response-4 (Par-4) is a unique pro-apoptotic molecule that is capable of selectively inducing apoptosis in cancer cells when over-expressed, sensitizing the cells to diverse apoptotic stimuli and causing regression of tumors in animal models. This review discusses the salient functions of Par-4 that can be harnessed to prostate cancer therapy. PMID- 14755682 TI - Approaches to understanding the importance and clinical implications of peroxisome proliferator-activated receptor gamma (PPARgamma) signaling in prostate cancer. AB - The development and maintenance of the prostate are dependent upon a complex series of interactions occurring between the epithelial and stromal tissues (Hayward and Cunha [2000]: Radiol. Clin. N. Am. 38:1-14). During the process of prostatic carcinogenesis, there are progressive changes in the interactions of the nascent tumor with its surrounding stroma and extracellular matrix. These include the development of a reactive stromal phenotype and the possible promotion, by stromal cells, of epithelial proliferation and loss of differentiated function (Hayward et al. [1996]: Ann. N. Y. Acad. Sci. 784:50-62; Grossfeld et al. [1998]: Endocr. Related Cancer 5:253-270; Rowley [1998]: Cancer Metastasis Rev. 17:411-419; Tuxhorn et al. [2002]: Clin. Cancer Res. 8:2912 2923). Many molecules play an as yet poorly defined role in establishing and maintaining a growth quiescent glandular structure in the adult. Peroxisome proliferator-activated receptor gamma (PPARgamma) is a candidate regulator of prostatic epithelial differentiation and may play a role in restricting epithelial proliferation. PPARgamma agonists are relatively non-toxic and have been used with limited success to treat some prostate cancer patients. We would propose that a more complete understanding of PPARgamma biology, particularly in the context of appropriate stromal-epithelial and host-tumor interactions would allow for the selection of patients most likely to benefit from this line of therapy. In particular, it seems reasonable to suggest that the patients most likely to benefit may be those with relatively indolent low stage disease for whom this line of therapy could be a useful additive to watchful waiting. PMID- 14755683 TI - Tumor target prostate specific membrane antigen (PSMA) and its regulation in prostate cancer. AB - Prostate specific membrane antigen (PSMA), is a unique membrane bound glycoprotein, which is overexpressed manifold on prostate cancer as well as neovasculature of most of the solid tumors, but not in the vasculature of the normal tissues. This unique expression of PSMA makes it an important marker as well as a large extracellular target of imaging agents. PSMA can serve as target for delivery of therapeutic agents such as cytotoxins or radionuclides. PSMA has two unique enzymatic functions, folate hydrolase and NAALADase and found to be recycled like other membrane bound receptors through clathrin coated pits. The internalization property of PSMA leads one to consider the potential existence of a natural ligand for PSMA. In this review we have discussed the regulation of PSMA expression within the cells, and significance of its expression in prostate cancer and metastasis. PMID- 14755684 TI - GSTP1 CpG island hypermethylation as a molecular biomarker for prostate cancer. AB - Somatic hypermethylation of CpG island sequences at GSTP1, the gene encoding the pi-class glutathione S-transferase, appears to be characteristic of human prostatic carcinogenesis. To consider the potential utility of this epigenetic alteration as a biomarker for prostate cancer, we present here a comprehensive review of the literature describing somatic GSTP1 changes in DNA from prostate cells and tissues. GSTP1 CpG island hypermethylation has been detected in prostate cancer DNA using a variety of assay techniques, including (i) Southern blot analysis (SB), after treatment with (5-m)C-sensitive restriction endonucleases, (ii) the polymerase chain reaction, following treatment with (5 m)C-sensitive restriction endonucleases (RE-PCR), (iii) bisulfite genomic sequencing (BGS), and (iv) bisulfite modification followed by the polymerase chain reaction, using primers selective for target sequences containing (5-m)C (MSP). In the majority of the case series so far reported, GSTP1 CpG island hypermethylation was present in DNA from at least 90% of prostate cancer cases. When analyses have been carefully conducted, GSTP1 CpG island hypermethylation has not been found in DNA from normal prostate tissues, or from benign prostatic hyperplasia (BPH) tissues, though GSTP1 CpG island hypermethylation changes have been detected in DNA from candidate prostate cancer precursor lesions proliferative inflammatory atrophy (PIA) and prostatic intraepithelial neoplasia (PIN). Using PCR methods, GSTP1 CpG island hypermethylation has also been detected in urine, ejaculate, and plasma from men with prostate cancer. GSTP1 CpG island hypermethylation, a somatic epigenetic alteration, appears poised to serve as a molecular biomarker useful for prostate cancer screening, detection, and diagnosis. PMID- 14755685 TI - Prostate cancer association studies: pitfalls and solutions to cancer misclassification in the PSA era. AB - Widespread screening of American men for elevated PSA has changed the characteristics of prostate cancer cases in the U.S. The influence of the changed nature of prostate cancer cases in the PSA era and the need for careful consideration of who is a "case" and who is a "control" on the ability to detect associations of risk factors with prostate cancer in etiologic epidemiologic studies merits discussion. Issue 1: prostate cancer cases diagnosed in the PSA era are enriched with a pool of early lesions, which may differ in etiology, and are deficient in advanced lesions, which are the most likely to be the product of promotion and progression events. By admixing the two types of cases (i.e., imperfect specificity), the associations previously detected using epidemiologic designs when the majority of cases were clinically detected may no longer be apparent in the PSA era when the majority of cases are now detected in the pre clinical phase. Researchers must now tailor hypotheses such that they are testable using early stage cases or specifically augment the number of advanced cases when testing hypotheses related to extraprostatic growth and progression. Issue 2: even when controls are screened for elevated PSA to rule out the presence of prostate cancer, some proportion of those controls currently will have one or more foci of prostate cancer. The imperfect sensitivity of the PSA test coupled with diagnostic work-up may in part result from (a) lack of PSA elevation in some men with prostate cancer or (b) failure of biopsy to sample the tumor focus in men with elevated PSA. Misclassification of men with undetected prostate cancer as controls usually produces a bias that tends to deflate associations. Given this type of disease misclassification, whether an association still can be statistically detected depends on the extent of misclassification, the magnitude of the true association, the prevalence of the exposure in the true controls, and the sample size, although in general moderate nondifferential misclassification does not lead to profound attenuation. However, under the same scenario attenuation does not occur in cohort or case-cohort studies in which the rate or risk ratio (RR) is calculated. That prostate cancer cases diagnosed in the PSA era are enriched with early stage, minimally invasive disease in our opinion is likely to pose a far more serious obstacle to epidemiologic research on the etiology of clinically important prostate cancer than the issue of inclusion as controls some men who have undiagnosed prostate cancer because of imperfect sensitivity of PSA screening and biopsy sampling error. PMID- 14755686 TI - Characterization of DNA demethylation in normal and cancerous cell lines and the regulatory role of cell cycle proteins in human DNA demethylase activity. AB - DNA methylation/demethylation constitutes a major consequence in all biological processes involving transcription, differentiation, development, DNA repair, recombination, and chromosome organization. Our earlier studies established that demethylation of CpG rich sequence by human DNA demethylase activity (5 methylcytosine-DNA glycosylase (5MeC-DNA glycosylase)) resembles "base excision DNA repair activity" and creates single-strand breaks on DNA that is associated with proliferating cell nuclear antigen (PCNA). Here in this report, we have identified differential DNA demethylation targets (hemi-methylated vs. fully methylated) in normal cell lines and cancerous cell lines, and a shortened G(0)/G(1) resting time in cancerous cell lines than the normal cell lines. We have identified that in normal HFL1 fibroblast cell line, DNA demethylase activity targets hemi-methylated CpG specific sites on DNA. This normal cell line DNA demethylase activity associates with PCNA immune complex that is inhibited by CDKI proteins p21(waf1)/Gadd45alpha and Gadd45beta. While in cancerous LnCap and BT20 cell lines DNA demethylase activity targets fully-methylated CpG specific sites on DNA. This cancer cell line DNA demethylase activity is not associated with PCNA immune complex and is not inhibited by CDKI proteins p21(waf1)/Gadd45alpha and Gadd45beta. We have also identified that the fully methylated CpG specific DNA demethylase activity from cancerous cell lines to associate with p300/CBP protein. These significant observations of variable targets of DNA demethylation and alternate partner proteins for DNA demethylase activity in cancerous cell lines are discussed in terms of double-strand DNA breaks versus single-strand DNA breaks and their role in the exit of G(1)/G(2) cell cycle stages. Also, the inability of cell cycle regulatory proteins like PCNA, p21(waf1), and Gadd45 to control DNA demethylase activity in cancerous cell lines is discussed in terms of accelerated G(1)/G(2) cell cycle stage exit to facilitate unregulated cellular proliferation, loss of control of chromosomal organization, and the development of oncogenesis in cancerous cell lines. PMID- 14755687 TI - Angiopoietin 1, PDGF-B, and TGF-beta gene regulation in endothelial cell and smooth muscle cell interaction. AB - The vascular wall is mainly composed of endothelial cells (ECs) and smooth muscle cells (SMCs). The crosstalking between these two cell types is critical in the vascular maturation process. Genetic studies suggest that the Tie2/angiopoietin 1 (Ang1) pathway regulates vascular remodeling. However, the molecular mechanism is unclear. PDGF is a potent chemoattractant for SMCs, and TGF-beta regulates SMC differentiation. Here, we examined gene regulation. PDGF-B stimulation upregulated Ang1 expression in SMCs through the PI3K and PKC pathways. PDGF-B stimulation also produced an acute induction of TGF-beta expression in SMCs through the MAPK/ERK pathway. Interestingly, TGF-beta negatively regulated Ang1 expression induced by the PDGF-B stimulation in SMCs. Reciprocally, we observed that stimulation of ECs with either Ang1 or TGF-beta slightly downregulated PDGF expression. A combination of both TGF-beta with Ang1 produced much stronger downregulation of PDGF. Our data showed complex gene regulations that include both positive and negative regulations between ECs and SMCs to maintain vascular homeostasis. PMID- 14755688 TI - Hydrocortisone increases the rate of differentiation of cultured human osteoblasts. AB - Reduced bone formation is the main finding in glucocorticoid-induced osteoporosis. The aim of this study was to determine whether differentiation of cultured human osteoblasts is inhibited by high concentrations of hydrocortisone. We measured the levels of mRNAs for three markers of cellular differentiation, type 1 collagen (COL1), alkaline phosphatase (ALP), and osteocalcin (OC), in four lines of human osteoblasts from female donors cultured with doses of hydrocortisone from 0 microM to 4 microM. The change in ALP/COL1 mRNA ratio over a given time was used to determine the average rate of differentiation of the cells in a culture. Although basal expression profiles and their changes with time were different for the different cell lines, all cell lines showed a dose dependent rise in the rate of increase of ALP mRNA relative to COL1 mRNA. However, increase in OC mRNA with time, seen here only in young donor hOBs, was significantly inhibited by 4 microM hydrocortisone, indicating that hydrocortisone can inhibit OC expression while promoting cellular differentiation. The data suggest that increasing concentrations of glucocorticoid, including concentrations similar to plasma levels in patients receiving oral glucocorticoid therapy, increase the rate of cellular differentiation. PMID- 14755689 TI - Analysis of the role of the leucine zipper motif in regulating the ability of AFAP-110 to alter actin filament integrity. AB - AFAP-110 has an intrinsic ability to alter actin filament integrity as an actin filament crosslinking protein. This capability is regulated by a carboxy terminal leucine zipper (Lzip) motif. The Lzip motif facilitates self-association stabilizing the AFAP-110 multimers. Deletion of the Lzip motif (AFAP 110(Deltalzip)) reduces the stability of the AFAP-110 multimer and concomitantly increases its ability to crosslink actin filaments, in vitro, and to activate cSrc and alter actin filament integrity, in vivo. We sought to determine how the Lzip motif regulates AFAP-110 function. Substitution of the c-Fos Lzip motif in place of the AFAP-110 Lzip motif (AFAP-110(fos)) was predicted to preserve the alpha-helical structure while changing the sequence. To alter the structure of the alpha-helix, a leucine to proline mutation was generated in the AFAP-110 alpha-helical Lzip motif (AFAP-110(581P)), which largely preserved the sequence. The helix mutants, AFAP-110(Deltalzip), AFAP-110(fos), and AFAP-110(581P), demonstrated reduced multimer stability with an increased capacity to crosslink actin filaments, in vitro, relative to AFAP-110. An analysis of opposing binding sites indicated that the carboxy terminus/Lzip motif can contact sequences within the amino terminal pleckstrin homology (PH1) domain indicating an auto-inhibitory mechanism for regulating multimer stability and actin filament crosslinking. In vivo, only AFAP-110(Deltalzip) and AFAP-110(581P) were to activate cSrc and to alter cellular actin filament integrity. These data indicate that the intrinsic ability of AFAP-110 to crosslink actin filaments is dependent upon both the sequence and structure of the Lzip motif, while the ability of the Lzip motif to regulate AFAP-110-directed activation of cSrc and changes in actin filament integrity in vivo is dependent upon the structure or presence of the Lzip motif. We hypothesize that the intrinsic ability of AFAP-110 to crosslink actin filaments or activate cSrc are distinct functions. PMID- 14755690 TI - Distinctive regulation and function of PI 3K/Akt and MAPKs in doxorubicin-induced apoptosis of human lung adenocarcinoma cells. AB - Regulation and function of PI 3K/Akt and mitogen-activated protein kinases (MAPKs) in doxorubicin-induced cell death were investigated in human lung adenocarcinoma cells. Doxorubicin induced dose-dependent apoptosis of human lung adenocarcinoma NCI-H522 cells. Prior to cell death, both Akt and the MAPK family members (MAPKs: ERK1/2, JNK, and p38) were activated in response to the drug treatment. The kinetics of the inductions for Akt and MAPKs are, however, distinct. The activation of Akt was rapid and transient, activated within 30 min of drug addition, then declined after 3 h, whereas the activations of three MAPKs occurred later, 4 h after addition of the drug and sustained until cell death occurred. Inhibition of PI 3K/Akt activation had no effect on MAPKs' activation, suggesting that the two pathways are independently activated in response to the drug treatment. Inhibition of PI 3K/Akt and p38 accelerated and enhanced doxorubicin-induced cell death. On the contrary, inhibition of ERK1/2 or JNK had no apparent effect on the cell death. Taken together, these results suggest that PI 3K/Akt and MAPKs signaling pathways are all activated, but with distinct mechanisms, in response to doxorubicin treatment. Activation of PI 3K/Akt and p38 modulates apoptotic signal pathways and inhibits doxorubicin-induced cell death. These responses of tumor cells to cancer drug treatment may contribute to their drug resistance. Understanding of the mechanism and function of the responses will be beneficial for the development of novel therapeutic approaches for improvement of drug efficacy and circumvention of drug resistance. PMID- 14755691 TI - A LIM protein, Hic-5, functions as a potential coactivator for Sp1. AB - Hic-5 is a LIM protein with striking similarity to paxillin, and shuttles between focal adhesions and the nucleus. Our previous study suggested that Hic-5 participates in the transcriptional control of several genes such as the c-fos and p21 genes. In the present study, we examined the function of Hic-5 in the nucleus using the transcriptional promoter region of the p21 gene. When localized to the nucleus, Hic-5 was found to transactivate the p21 promoter through two of five Sp1 sites in the region proximal to the TATA box. The Hic-5 effect was mediated by a transactivation domain of Sp1 and functional interaction with p300 through the LIM4 domain. Hic-5 was also shown to interact functionally and physically with Smad3 through the LIM domains and to potentiate p21 promoter activity together with Smad3 and Sp1. These properties were confirmed in an artificial system using GAL4-fusion protein. Thus, Hic-5 was suggested to have a potential function as a cofactor in the transcriptional complex that contains Sp1, playing a role in gene transcription in the nucleus as well as in integrin signaling at focal adhesion sites. PMID- 14755694 TI - Novel compounds of elements of group 14: ligand-stabilized clusters with "naked" atoms. PMID- 14755695 TI - G-quartets 40 years later: from 5'-GMP to molecular biology and supramolecular chemistry. AB - Molecular self-assembly is central to many processes in both biology and supramolecular chemistry. The G-quartet, a hydrogen-bonded macrocycle formed by cation-templated assembly of guanosine, was first identified in 1962 as the basis for the aggregation of 5'-guanosine monophosphate. We now know that many nucleosides, oligonucleotides, and synthetic derivatives form a rich array of functional G-quartets. The G-quartet surfaces in areas ranging from structural biology and medicinal chemistry to supramolecular chemistry and nanotechnology. This Review integrates and summarizes knowledge gained from these different areas, with emphasis on G-quartet structure, function, and molecular recognition. PMID- 14755696 TI - A crystalline microporous narrow-bandgap semiconductor. PMID- 14755697 TI - Supercritical-fluid processing technique for nanoscale polymer particles. PMID- 14755698 TI - A three-dimensional ferrimagnet composed of mixed-valence Mn4 clusters linked by an [Mn[N(CN)2]6]4- unit. PMID- 14755699 TI - Catalytic generation of indium hydride in a highly diastereoselective reductive aldol reaction. PMID- 14755700 TI - The circumambulation of a phosphirane: taking 9-phenyl-9 phosphabicyclo[6.1.0]nona-2,4,6-triene for a "walk". PMID- 14755701 TI - Synthesis of thermoresponsive polyurethane from 2-methylaziridine and supercritical carbon dioxide. PMID- 14755702 TI - Tandem 1,4-addition/enantioselective protonation catalyzed by rhodium complexes: efficient access to alpha-amino acids. PMID- 14755703 TI - A peptide flavoprotein mimic: flavin recognition and redox potential modulation in water by a designed beta hairpin. PMID- 14755704 TI - Synthesis and incorporation of the first polyketide synthase free intermediate in monocerin biosynthesis. PMID- 14755705 TI - Oxidative polymerization of 2,6-dimethylphenol to form poly(2,6-dimethyl-1,4 phenyleneoxide) in water. PMID- 14755706 TI - Electrochemical preparation of silicon and its alloys from solid oxides in molten calcium chloride. PMID- 14755707 TI - Synthesis of a delocalized azepinium ion and investigation of its electrophilic character. PMID- 14755708 TI - Stereoselective total synthesis of (+)-norrisolide. PMID- 14755709 TI - 5-Dehydro-1,3-quinodimethane: a hydrocarbon with an open-shell doublet ground state. PMID- 14755710 TI - The direct synthesis of a silene-organometallic complex. PMID- 14755711 TI - High-throughput synthesis of phosphonate-based inorganic-organic hybrid compounds under hydrothermal conditions. PMID- 14755712 TI - High-throughput method for the impedance spectroscopic characterization of resistive gas sensors. PMID- 14755713 TI - Superphenalene-based columnar liquid crystals. PMID- 14755715 TI - Alpha-synuclein in Parkinson's disease: light from two new angles. PMID- 14755716 TI - Dissecting the relative influences of genes and the environment in Alzheimer's disease. PMID- 14755717 TI - Biomarker epidemiology of cerebral palsy. PMID- 14755718 TI - Why muscle atrophy in acute quadriplegic myopathy is rapid and severe. PMID- 14755719 TI - The new mutation, E46K, of alpha-synuclein causes Parkinson and Lewy body dementia. AB - Familial parkinsonism and dementia with cortical and subcortical Lewy bodies is uncommon, and no genetic defect has been reported in the previously described sibships. We present a Spanish family with autosomal dominant parkinsonism, dementia, and visual hallucinations of variable severity. The postmortem examination showed atrophy of the substantia nigra, lack of Alzheimer pathology, and numerous Lewy bodies which were immunoreactive to alpha-synuclein and ubiquitin in cortical and subcortical areas. Sequencing of the alpha-synuclein gene showed a novel, nonconservative E46K mutation in heterozygosis. The E46K mutation was present in all affected family members and in three young asymptomatic subjects, but it was absent in healthy and pathological controls. The novel mutation, that substitutes a dicarboxylic amino acid, glutamic acid, with a basic amino acid such as lysine in a much conserved area of the protein, is likely to produce severe disturbance of protein function. Our data show that, in addition to the previously described hereditary alpha-synucleinopathies, dementia with Lewy bodies is related to mutation of alpha-synuclein. PMID- 14755720 TI - Comparison of kindreds with parkinsonism and alpha-synuclein genomic multiplications. AB - Genomic triplication of the alpha-synuclein gene recently has been associated with familial Parkinson's disease in the Spellman-Muenter kindred. Here, we present an independent family, of Swedish-American descent, with hereditary early onset parkinsonism with dementia due to alpha-synuclein triplication. Brain tissue available from affected individuals in both kindreds provided the opportunity to compare their clinical, pathological, and biochemical phenotypes. Of note, studies of brain mRNA and soluble protein levels demonstrate a doubling of alpha-synuclein expression, consistent with molecular genetic data. Pathologically, cornu ammonis 2/3 hippocampal neuronal loss appears to be a defining feature of this form of inherited parkinsonism. The profound implications of alpha-synuclein overexpression for idiopathic synucleinopathies are discussed. PMID- 14755721 TI - How heritable is Alzheimer's disease late in life? Findings from Swedish twins. AB - Although genetic effects are known to be important for early onset Alzheimer's disease, little is known about the importance of genetic effects for late-onset disease. Furthermore, previous studies are based on prevalent cases. Our purpose was to characterize the relative importance of genetic and environmental factors for incident Alzheimer's disease late in life, and to test for differences in the importance of genetic effects at different ages. A cohort of 662 pairs of Swedish twins 52 to 98 years of age who were without symptoms of dementia was followed up for an average of 5 years. Incident dementia cases were detected through follow up at 2 to 3-year intervals using either cognitive testing or telephone screening followed by dementia workups. A physician, psychologist, and nurse gave consensus diagnoses. During the follow-up period, 5.8% of the sample was diagnosed with Alzheimer's disease. Average age of onset was 83.9 years (standard deviation, 6.3). Of the 26 monozygotic pairs in which at least one twin developed Alzheimer's disease, 5 were concordant (probandwise concordance, 32.2%). The concordance rate for dizygotic pairs was 8.7% (2 of 44 pairs). Structural model fitting indicated that 48% of the variation in liability to Alzheimer's disease could be attributed to genetic variation. Estimates did not differ significantly between twins younger than age 80 years and those older than age 80 years at baseline. Although these genetic estimates for incident disease are lower than those for prevalent disease, the importance of genetic factors for liability to Alzheimer's disease is considerable even late in life. PMID- 14755722 TI - Cerebral palsy is characterized by protein mediators in cord serum. AB - Cerebral palsy (CP) is a major neurodevelopmental disability in childhood. An association between intrauterine infection and CP has been reported. We examined the relationship between inflammatory mediators in cord serum and CP in term and preterm children. Regional multicenter study was conducted on 19 CP children and 19 gestation-matched paired controls. CP children (n = 27) were further compared with controls of similar gestation at birth (n = 25). Serum levels of 78 protein mediators were analyzed. Eleven analytes correlated with the length of gestation both in cases and controls. In paired analysis, B-lymphocyte chemoattractant, ciliary neurotrophic factor, epidermal growth factor, interleukin (IL)-5, IL-12, IL-13, IL-15, macrophage migration inhibitory factor, monocyte chemoattractant protein-3, monokine induced by interferon gamma, and tumor necrosis factor related apoptosis-inducing ligand were higher in children with CP (p < or = 0.05). Preterm infants with CP showed higher epidermal growth factor and lower levels of granulocyte-macrophage colony-stimulating factor, IL-2, macrophage derived chemokine, and pulmonary and activation-regulated chemokine than their paired controls. Inflammatory mediators and growth factors serve as a footprint of the fetal response to an insult manifesting after birth as a permanent brain damage. The cytokine patterns at birth differ between premature and term infants who develop CP. PMID- 14755723 TI - Constitutive activation of MAPK cascade in acute quadriplegic myopathy. AB - Acute quadriplegic myopathy (AQM; also called "critical illness myopathy") shows acute muscle wasting and weakness and is experienced by some patients with severe systemic illness, often associated with administration of corticosteroids and/or neuroblocking agents. Key aspects of AQM include muscle atrophy and myofilament loss. Although these features are shared with neurogenic atrophy, myogenic atrophy in AQM appears mechanistically distinct from neurogenic atrophy. Using muscle biopsies from AQM, neurogenic atrophy, and normal controls, we show that both myogenic and neurogenic atrophy share induction of myofiber-specific ubiquitin/proteosome pathways (eg, atrogin-1). However, AQM patient muscle showed a specific strong induction of transforming growth factor (TGF)-beta/MAPK pathways. Atrophic AQM myofibers showed coexpression of TGF-beta receptors, p38 MAPK, c-jun, and c-myc, including phosphorylated active forms, and these same fibers showed apoptotic features. Our data suggest a model of AQM pathogenesis in which stress stimuli (sepsis, corticosteroids, pH imbalance, osmotic imbalance) converge on the TGF-beta pathway in myofibers. The acute stimulation of the TGF beta/MAPK pathway, coupled with the inactivity-induced atrogin-1/proteosome pathway, leads to the acute muscle loss seen in AQM patients. PMID- 14755724 TI - Characterizing the diffusion/perfusion mismatch in experimental focal cerebral ischemia. AB - Diffusion-weighted imaging (DWI) and perfusion-weighted imaging (PWI) can rapidly detect lesions in acute ischemic stroke patients. The PWI volume is typically substantially larger than the DWI volume shortly after onset, that is, a diffusion/ perfusion mismatch. The aims of this study were to follow the evolution of the diffusion/ perfusion mismatch in permanent and 60- minute temporary focal experimental ischemia models in Sprague-Dawley rats using the intraluminal middle cerebral artery occlusion (MCAO) method. DWI and arterial spin-labeled PWI were performed at 30, 60, 90, 120, and 180 minutes after occlusion and lesion volumes (mm(3)) calculated At 24 hours after MCAO, and infarct volume was determined using triphenyltetrazolium chloride staining. In the permanent MCAO group, the lesion volume on the ADC maps was significantly smaller than that on the cerebral blood flow maps through the first 60 minutes after MCAO; but not after 90 minutes of occlusion. With 60 minutes of transient ischemia, the diffusion/perfusion mismatch was similar, but after reperfusion, the lesion volumes on ADC and cerebral blood flow maps became much smaller. There was a significant difference in 24- hour infarct volumes between the permanent and temporary occlusion groups. PMID- 14755725 TI - Clinical features of cerebral cavernous malformations patients with KRIT1 mutations. AB - Cerebral Cavernous Malformations (CCM/OMIM 604214) are vascular malformations causing seizures and cerebral hemorrhages. They occur as a sporadic and autosomal dominant condition, the latter being characterized by the presence of multiple CCM lesions. Stereotyped truncating mutations of KRIT1, the sole CCM gene identified so far, have been identified in CCM1 linked families but the clinical features associated with KRIT1 mutations have not yet been assessed in a large series of patients. We conducted a detailed clinical, neuroradiological and molecular analysis of 64 consecutively recruited CCM families segregating a KRIT1 mutation. Those families included 202 KRIT1 mutation carriers. Among the 202 KRIT1 mutation carriers, 126 individuals were symptomatic and 76 symptom-free. Mean age at clinical onset was 29.7 years (range, 2-72); initial clinical manifestations were seizures in 55% of the cases and cerebral hemorrhages in 32%. Average number of lesions on T2 weighted MRI was 4.9 (+/-7.2) and on gradient echo sequences 19.8 (+/-33.2). Twenty-six mutation carriers harbored only one lesion on T2-weighted MRI, including 4 mutation carriers, aged from 18 to 55 yr old, who presented only one CCM lesion both on T2-weighted and on highly sensitive gradient echo MRI sequences. Five symptom free mutation carriers, aged from 27 to 48 yr-old, did not have any detectable lesion both on T2WI and gradient echo MRI sequences. Within KRIT1/CCM1 families, both clinical and radiological penetrance are incomplete and age dependent. Importantly for genetic counseling, nearly half of the KRIT1 mutation carriers aged 50 or more are symptom-free. The presence of only one lesion, even when using gradient echo MRI sequences, can be observed in some patients with an hereditary form of the disease. Incomplete neuroradiological penetrance precludes the use of cerebral MRI to firmly establish a non carrier status, even at an adult age and even when using highly sensitive gradient echo MRI. Altogether these data suggest that the hereditary nature of the disorder may be overlooked in some mutation carriers presenting as sporadic cases with a unique lesion. PMID- 14755726 TI - Presence of dendritic cells, MCP-1, and activated microglia/macrophages in amyotrophic lateral sclerosis spinal cord tissue. AB - Dendritic cells are potent antigen-presenting cells that initiate and amplify immune responses. To determine whether dendritic cells participate in inflammatory reactions in amyotrophic lateral sclerosis (ALS), we examined mRNA expression of dendritic cell surface markers in individual sporadic ALS (sALS), familial ALS (fALS), and nonneurological disease control (NNDC) spinal cord tissues using semiquantitative and real-time reverse transcription polymerase chain reaction (RT-PCR). Immature (DEC205, CD1a) and activated/mature (CD83, CD40) dendritic cell transcripts were significantly elevated in ALS tissues. The presence of immature and activated/mature dendritic cells (CD1a(+) and CD83(+)) was confirmed immunohistochemically in ALS ventral horn and corticospinal tracts. Monocytic/macrophage/microglial transcripts (CD14, CD18, SR-A, CD68) were increased in ALS spinal cord, and activated CD68(+) cells were demonstrated in close proximity to motor neurons. mRNA expressions of the chemokine MCP-1, which attracts monocytes and myeloid dendritic cells, and of the cytokine macrophage colony stimulating factor (M-CSF) were increased in ALS tissues. The MCP-1 protein was expressed in glia in ALS but not in control tissues and was increased in the CSF of ALS patients. Those patients who progressed most rapidly expressed significantly more dendritic transcripts than patients who progressed more slowly. These results support the involvement of immune/inflammatory responses in amplifying motor neuron degeneration in ALS. PMID- 14755727 TI - Somatic mutations in VHL germline deletion kindred correlate with mild phenotype. AB - Generally, von Hippel-Lindau (VHL) disease is caused by a germline mutation of the VHL gene (chromosome 3p), and tumorigenesis is initiated from a "second-hit" deletion. A subset of VHL patients have a germline deletion of the VHL gene, and the molecular events leading to tumorigenesis are not fully understood. To determine the molecular pathogenesis of tumor formation in this setting, we analyzed five central nervous system hemangioblastomas from three patients of a single VHL germline deletion kindred, all displaying mild clinical phenotype. Rather than loss of heterozygosity (the "second hit" in VHL germline mutation patients), all tumors from this kindred showed "second-hit" point mutations on the wild-type allele. Moreover, in two patients who each had two hemangioblastomas resected each tumor contained a unique mutation. The specific germline deletion and the overall genetic makeup of the patient did not predict these random "second-hit" point mutations. These results suggest that in patients with germline deletion of a tumor suppressor gene there is a unique genetic mechanism underlying tumorigenesis. This unique genetic mechanism correlates with and may help to understand the mild clinical phenotype seen in these patients. PMID- 14755728 TI - Aprataxin, the causative protein for EAOH is a nuclear protein with a potential role as a DNA repair protein. AB - Early-onset ataxia with ocular motor apraxia and hypoalbuminemia (EAOH) is an autosomal recessive neurodegenerative disorder characterized by early-onset ataxia, ocular motor apraxia, and hypoalbuminemia. Recently, the causative gene for EAOH, APTX, has been identified. Of the two splicing variants of APTX mRNA, the short and the long forms, long-form APTX mRNA was found to be the major isoform. Aprataxin is mainly located in the nucleus, and, furthermore, the first nuclear localization signal located near the amino terminus of the long-form aprataxin is essential for its nuclear localization. We found, based on the yeast two-hybrid and coimmunoprecipitation experiments, that the long-form but not the short-form aprataxin interacts with XRCC1 (x-ray repair cross-complementing group 1). Interestingly the amino terminus of the long-form aprataxin is homologous with polynucleotidekinase-3'-phosphatase, which has been demonstrated to be involved in base excision repair, a subtype of single-strand DNA break repair, through interaction with XRCC1, DNA polymerase beta, and DNA ligase III. These results strongly support the possibility that aprataxin and XRCC1 constitute a multiprotein complex and are involved in single-strand DNA break repair, and furthermore, that accumulation of unrepaired damaged DNA underlies the pathophysiological mechanisms of EAOH. PMID- 14755729 TI - Clinical manifestations of cerebral amyloid angiopathy-related inflammation. AB - To explore the clinical effects of inflammation associated with vascular deposits of the amyloid beta peptide (A beta), we analyzed 42 consecutive patients with pathologically diagnosed cerebral amyloid angiopathy (CAA) for evidence of an inflammatory response. Inflammation with giant-cell reaction surrounding amyloid laden vessels was identified in 7 of the 42 cases. The clinical symptoms in each of the seven were subacute cognitive decline or seizure rather than hemorrhagic stroke, the primary clinical presentation in 33 of 35 patients with noninflammatory CAA (p < 0.001). Inflammatory CAA also was associated with radiographic white matter abnormalities, significantly younger age at presentation, and a marked overrepresentation of the apolipoprotein E epsilon 4/epsilon 4 genotype (71% vs 4%, p < 0.001). Of the six inflammatory CAA patients with available follow-up information, five demonstrated clinical and radiographic improvement after immunosuppressive treatment. The syndrome of CAA-related perivascular inflammation appears to represent a subset of CAA with clinically distinct symptoms that may respond to immunosuppressive treatment. These data add to evidence that inflammation against A beta can cause vascular dysfunction, a potential mechanism for the toxic response recently observed in clinical trials of A beta immunization. PMID- 14755730 TI - Perturbation of sphingolipid metabolism and ceramide production in HIV-dementia. AB - Infection by the human immunodeficiency virus type 1 (HIV-1) often results in neurological dysfunction including HIV dementia (HIVD). Alterations in cytokine and redox balance are thought to play important roles in the pathogenesis of HIVD, but the specific mechanisms underlying neuronal dysfunction and death are unknown. Activation of cytokine receptors and oxidative stress can induce the production of ceramide from membrane sphingomyelin, and recent findings suggest that ceramide is an important mediator of a form of programmed cell death called apoptosis. We now report that levels of ceramide, sphingomyelin, and hydroxynonenal (HNE) are significantly increased in brain tissues and cerebrospinal fluid of HIVD patients. Exposure of cultured neurons to the neurotoxic HIV proteins gp120 and Tat resulted in increased cellular levels of sphingomyelin, ceramide, and HNE. The ceramide precursor palmitoyl-CoA sensitized neurons to Tat and gp120 toxicity, whereas an inhibitor of ceramide production reduced Tat and gp120-induced increases of ceramide and HNE and protected the neurons from Tat and gp120-induced death. These results suggest that HIV-1 infection may promote a lipid imbalance in neural cells, resulting in an overproduction of ceramide and consequent cellular dysfunction and death. PMID- 14755731 TI - Deterioration of naming nouns versus verbs in primary progressive aphasia. AB - Disproportionate impairment of naming nouns versus verbs and the opposite pattern have been reported in cases of focal brain damage or degenerative disease, indicating that processing of nouns and verbs may rely on different brain regions. However, it has not been clear whether it is the spoken word forms or the meanings (or both) of nouns and verbs that depend on separate neural regions. We tested oral and written naming of nouns and verbs, matched in difficulty, in patients with nonfluent primary progressive aphasia (nonfluent PPA; n = 15), fluent primary progressive aphasia (fluent PPA; n = 7), and amyotrophic lateral sclerosis with frontotemporal dementia (ALS-FTD; n = 6). Patients with nonfluent PPA and ALS-FTD, both individually and as groups, were significantly more impaired on verb naming than on noun naming and significantly more impaired on oral naming than written naming. Patients with fluent PPA showed the opposite pattern for both word class and modality, significantly more impaired naming of nouns versus verbs and significantly more impaired written versus oral naming. Results indicate that separate regions of the brain are essential for access to oral and written word forms of verbs and nouns, and that these neural regions can be differentially damaged in separate forms of PPA. PMID- 14755732 TI - Deciphering migraine mechanisms: clues from familial hemiplegic migraine genotypes. PMID- 14755733 TI - SCA17 homozygote showing Huntington's disease-like phenotype. AB - We report a homozygous case of spinocerebellar ataxia type 17 with 48 glutamines. The age of the patient at disease onset was not lower than those of heterozygotes with the same CAG-repeat sizes, but the clinical manifestations were rapidly progressive dementia and chorea. Neuronal loss was relatively restricted and most prominent in the Purkinje cell layer and striatum; however, intranuclear neuronal polyglutamine accumulation was widespread, with a high frequency in the cerebral cortex and striatum. PMID- 14755734 TI - Rhabdomyolysis and paraneoplastic stiff-man syndrome with amphiphysin autoimmunity. AB - Stiff-Man syndrome (SMS) is a rare disease of the central nervous system characterized by chronic muscle rigidity and autoimmunity directed against synaptic antigens. In a subset of patients, generally positive for antiamphiphysin autoantibodies, SMS has an autoimmune paraneoplastic origin. Amphiphysin isoforms are expressed at high levels in brain and skeletal muscle and often are overexpressed in breast cancer. We report here the occurrence of rhabdomyolysis in a patient with SMS, breast cancer, and antibodies that recognize both brain and muscle amphiphysin isoforms. Immunotherapy induced a remission of both rhabdomyolysis and SMS symptoms. Autoimmune rhabdomyolysis may represent a paraneoplastic complication of cancer patients with amphiphysin autoimmunity. PMID- 14755735 TI - Autosomal dominant acute necrotizing encephalopathy maps to 2q12.1-2q13. AB - In autosomal dominant acute necrotizing encephalopathy (ADANE), apparently healthy children develop necrotizing lesions in their thalami and brainstems in the course of febrile illnesses. We used DNA from affected subjects and obligate carriers to map ADANE to a 6.5Mb region on chromosome 2. Sequencing of four candidate genes in the interval (BCL2L11, ST6GalII, CHT1, and FLJ20019), involved in apoptosis, viral recognition, choline transport, and electron transport, showed no disease causing mutations. PMID- 14755736 TI - Prion deposition in olfactory biopsy of sporadic Creutzfeldt-Jakob disease. AB - Currently, definite peripheral markers for the in vivo diagnosis of sporadic Creutzfeldt-Jakob disease (CJD) are not available. Here, we report the presence of pathological prion protein in the olfactory mucosa of a case with sporadic Creutzfeldt-Jakob disease. Prion protein immunoreactivity was detected in an olfactory biopsy performed 45 days after the disease onset, suggesting that the involvement of olfactory epithelium is an early event in sporadic Creutzfeldt Jakob disease. PMID- 14755737 TI - More transgenic mouse models of dopamine deficiency. PMID- 14755738 TI - Multiple sclerosis: should MR criteria for dissemination in time be less stringent. PMID- 14755739 TI - Disease penetrance in amyotrophic lateral sclerosis associated with mutations in the SOD1 gene. PMID- 14755740 TI - Herpes simplex virus type 1 and Alzheimer's disease. PMID- 14755747 TI - Anatomy and function of the ptychoid defensive mechanism in the mite Euphthiracarus cooki (Acari: Oribatida). AB - Ptychoidy is a defensive adaptation of several groups of oribatid mites in which legs and coxisternum can be fully retracted into the opisthosoma and protected by a ventrally deflected prodorsum, resulting in a seed-like appearance. Using Euphthiracarus cooki as a model, we examined details of exoskeletal and muscular anatomy in combination with studies of live individuals to provide the first functional analysis of ptychoidy. There are two main functional components: the first is a set of exoskeletal and muscular adaptations, mostly of the podosoma and prodorsum, that combine to effect leg withdrawal and prodorsal deflection; the second comprises adaptations of the opisthosoma that allow control of hydrostatic pressure during the large hemocoel volume adjustments associated with ptychoidy. Adaptations important in the closing process (enptychosis) are found in four body regions. Much of the podosomal exoskeleton (especially pleural) is unsclerotized, which facilitates leg retraction and prodorsal deflection during enptychosis. The coxisternum has several flexible furrows along which it folds in order to bring legs into a tightly parallel arrangement. The prodorsum has specialized attachment surfaces (manubrium and inferior retractor process) for retractor muscles and a paired bothridial scale that participates in prodorsal alignment during enptychosis. The subcapitulum has a prominent capitular apodeme on which important retractor muscles insert. The mineralized notogaster has an anterior "collar" that accommodates the retracted prodorsum; it includes paired notches and receptacles that accommodate the bothridial scales, thereby creating a temporary fixed axis for rotation of the prodorsum in a "lazy hinge" mechanism. Specialized muscles form the retractor system; most conspicuous are the large coxisternal retractors and prodorsal retractors, both of which originate on the notogaster. Other components have adjustor roles; among them are muscles of the endosternal system that control retraction of the subcapitulum and assist leg retraction, and the dorsoventral muscles which adjust the folded coxisternum. Hemolymph pressure control is a function of the opisthosoma, where the principle exoskeletal elements form a pleated venter, having a cross-sectional shape like an inverted "W." Paired holoventral plates (each representing fused genital, aggenital, anal, and anal plates) form the inner angle and are flanked by paired plicature plates. The holoventral plates are connected medially in two ways : 1) by three permanent bridges of sclerotized cuticle that include an anterior phragmatal bridge and two widely spaced, hollow apodemes (preanal, postanal); 2) by temporary zipper-like closures of two different types. Lateral compression of the pleats is effected by a series of transversely arranged, lateral compressor muscles that run from plicature to holoventral plate edges, and from holoventral plate edges to the medial apodemes. Compression increases hydrostatic pressure in the opisthosoma and stores energy in both the slightly deformed, mineralized notogaster and in the three holoventral bridges. During normal activity the compressor system is active and the inflated podosomal region provides support for the extended legs. When the mite is irritated, the prodorsum is hydraulically ejected from its active position in the notogastral collar, then relaxation of the compressors causes a fall in hemolymph pressure and return of the notogaster to an undeformed condition. Muscles of the retractor system then act in specific sequence to retract and adjust the coxisternum and prodorsum until they are precisely positioned at the completion of enptychosis. The process takes between 0.5 and 1 sec. When irritation ceases, partial opening allows sensory leg hairs to "test" the environment. Resumption of normal, active posture (ecptychosis) involves activation of the lateral compressor system and hydraulic inflation of the podosoma, through which legs are extended and the prodorsum is reflected. PMID- 14755748 TI - From head to tail: the myoseptal system in basal actinopterygians. AB - Experimental studies indicated that myomeres play several functional roles during swimming. Some of the functions in question are thought to change rostrocaudally, e.g., anterior myomeres are thought to generate forces, whereas posterior myomeres are thought to transmit forces. In order to determine whether these putative functions are reflected in myoseptal morphology we carried out an analysis of the myoseptal system that includes epaxial and hypaxial myosepta of all body regions for the first time. We combined clearing and staining, microdissections, polarized light microscopy, SEM technique, and length measurements of myoseptal parts to reveal the spatial arrangement, collagen fiber architecture, and rostrocaudal gradients of myosepta. We included representatives of the four basal actinopterygian clades to evaluate our findings in an evolutionary and in a functional context. Our comparison revealed a set of actinopterygian groundplan features. This includes a set of specifically arranged myoseptal tendons (epineural, epipleural, lateral, and myorhabdoid tendons) in all epaxial and postanal hypaxial myosepta. Only preanal hypaxial myosepta lack tendons and exclusively consist of mediolateral fibers. Laterally, myosepta generally align with the helically wound fibers of the dermis in order not to limit the body's maximum curvature. Medially, the relationship of myosepta to vertebrae clearly differs from a 1:1 relationship: a myoseptum attaches to the anterior margin of a vertebra, turns caudally, and traverses at least three vertebrae in an almost horizontal orientation in all body regions. By this arrangement, horizontal multiple layers of myosepta are formed along the trunk dorsal and ventral to the horizontal septum. Due to their reinforcement by epineural or epipleural tendons, these multiple layers are hypothesized to resist the radial expansion of underlying muscle fibers and thus contribute to modulation of body stiffness. Rostrocaudally, a dorsoventral symmetry of epaxial and hypaxial myosepta in terms of spatial arrangement and collagen fiber architecture is gradually developed towards the postanal region. Furthermore, the rostrocaudal extension of myosepta measured between anterior and posterior cones gradually increases. This myoseptal region is reinforced by longitudinal fibers of lateral tendons. Furthermore, the percentage of connective tissue in a cross section increases. These morphological data indicate that posterior myosepta are equipped for multisegmental force transmission towards the caudal fin. Anteriormost myosepta have reinforced and elongated dorsal posterior cones. They are adequately designed to transmit epaxial muscular forces to the neurocranium in order to cause its elevation during suction feeding. PMID- 14755749 TI - Sexual dimorphism in the craniofacial growth of the guinea pig (Cavia porcellus). AB - Variation between the sexes during ontogeny is frequently overlooked in discussions of the phylogenetic patterns of adult sexual dimorphism. Different growth trajectories can produce identical degrees and direction of adult dimorphism and the possibility exists that similarities in adults may be the result of differing growth patterns, suggesting independent evolutionary pathways among species to the seemingly identical adult morphology. We quantified the sexual dimorphism in craniofacial skeletal growth of Cavia porcellus, the guinea pig, using longitudinally collected radiographs. Guinea pigs have male-biased sexual dimorphism in size and in growth parameters, despite literature reports to the contrary. These results, analyzed with equivalent data for five species of rodents, and two outgroups representing similarly sized mammals, a rabbit and a marsupial, indicate that some aspects of sexual differences in growth follow phylogenetic lines, while others are a function of whether the species has male- or female-biased dimorphism. PMID- 14755750 TI - Synthesis of interkeratin matrix in differentiating lizard epidermis: an ultrastructural autoradiographic study after injection of tritiated proline and histidine. AB - During epidermal differentiation in mammals, keratins and keratin-associated matrix proteins rich in histidine are synthesized to produce a corneous layer. Little is known about interkeratin proteins in nonmammalian vertebrates, especially in reptiles. Using ultrastructural autoradiography after injection of tritiated proline or histidine, the cytological process of synthesis of beta keratin and interkeratin material was studied during differentiation of the epidermis of lizards. Proline is mainly incorporated in newly synthesized beta keratin in beta-cells, and less in oberhautchen cells. Labeling is mainly seen among ribosomes within 30 min postinjection and appears in beta-keratin packets or long filaments 1-3 h later. Beta-keratin appears as an electron-pale matrix material that completely replaces alpha-keratin filaments in cells of the beta layer. Tritiated histidine is mainly incorporated into keratohyalin-like granules of the clear layer, in dense keratin bundles of the oberhautchen layer, and also in dense keratin filaments of the alpha and lacunar layer. The detailed ultrastructural study shows that histidine-labeling is localized over a dense amorphous material associated with keratin filaments or in keratohyalin-like granules. Large keratohyalin-like granules take up labeled material at 5-22 h postinjection of tritiated histidine. This suggests that histidine is utilized for the synthesis of keratins and keratin-associated matrix material in alpha keratinizing cells and in oberhautchen cells. As oberhautchen cells fuse with subjacent beta-cells to form a syncytium, two changes occur : incorporation of tritiated histidine, but uptake of proline increases. The incorporation of tritiated histidine in oberhautchen cells lowers after merging with cells of the beta-layer, whereas instead proline uptake increases. In beta-cells histidine labeling is lower and randomly distributed over the cytoplasm and beta-keratin filaments. Thus, change in histidine uptake somehow indicates the transition from alpha- to beta-keratogenesis. This study indicates that a functional stratum corneum in the epidermis of amniotes originates only after the association of matrix and corneous cell envelope proteins with the original keratin scaffold of keratinocytes. PMID- 14755751 TI - Phylogenetic comparison of serotonin-immunoreactive neurons in representatives of the Chilopoda, Diplopoda, and Chelicerata: implications for arthropod relationships. AB - The phylogenetic relationships within the Arthropoda have been discussed controversially for more than a century. Comparative studies on structure and development of the nervous system have contributed important arguments to this discussion. Arthropods have individually identifiable neurons that can be used as characters in phylogenetic studies. In the present report, the arrangement of serotonin-immunoreactive neurons in the ventral nerve cord was examined in seven representatives of the Chelicerata, Chilopoda, and Diplopoda. The goal of this analysis was to determine whether number, arrangement, and axonal morphology of the serotonergic neurons in these groups are similar to the pattern found in representatives of the Hexapoda and Crustacea, as explored in a previous study. The results indicate that the pattern in the seven species examined here does not correspond to that present in the Hexapoda and Crustacea. In particular, the pattern in Chilopoda and Diplopoda is clearly different from that of the Hexapoda. The hexapodan pattern most closely resembles that of the Crustacea. These findings are discussed with regard to recent reports on the mechanisms of neurogenesis in these taxa. Furthermore, the proposed ground patterns of the various groups are reconstructed and the characters are plotted on two competing hypotheses of arthropod phylogeny, the traditional Tracheata hypothesis and an alternative hypothesis derived from molecular and recent morphological data, the Tetraconata concept. The data discussed in this article moderately support the Tetraconata hypothesis. PMID- 14755752 TI - Placental ontogeny of the Tasmanian scincid lizard, Niveoscincus ocellatus (Reptilia: Squamata). AB - A prominent scenario for the evolution of reptilian placentation infers that placentotrophy arose by gradual modification of a simple vascular chorioallantoic placenta to a complex structure with a specialized region for nutrient transfer. The structure of the chorioallantoic placenta of Niveoscincus ocellatus, apparently described originally from a single embryonic stage, was interpreted as a transitional evolutionary type that provided support for the model. Recently, N. ocellatus has been found to be as placentotrophic as species with complex chorioallantoic placentae containing a specialized region called a placentome. We studied placental development in N. ocellatus and confirmed that the chorioallantoic placenta lacks specializations found in species with a placentome. We also found that this species has a specialized omphaloplacenta. The chorioallantoic placenta is confined to the region adjacent to the embryo by a membrane, similar to that found in some other viviparous skinks, that divides the egg into embryonic and abembryonic hemispheres. We term this structure the "inter-omphalopleuric" membrane. The position of this membrane in N. ocellatus is closer to the embryonic pole of the egg than to the abembryonic pole and thus the surface area of the omphaloplacenta is greater than that of the chorioallantoic placenta. In addition, the omphaloplacenta is regionally diversified and more complex histologically than the chorioallantoic placenta. An impressive and unusual feature of the omphaloplacenta of N. ocellatus is the development of extensive overlapping folds in the embryonic component of mid-gestation embryos. The histological complexity and extensive folding of the omphaloplacenta make this a likely site of placental transfer of nutrients in this species. PMID- 14755753 TI - Immunocytochemical and autoradiographic studies on the process of keratinization in avian epidermis suggests absence of keratohyalin. AB - The process of keratinization in apteric avian epidermis and in scutate scales of some avian species has been studied by autoradiography for histidine and immunohistochemistry for keratins and other epidermal proteins. Acidic or basic alpha-keratins are present in basal, spinosus, and transitional layers, but are not seen in the corneous layer. Keratinization-specific alpha-keratins (AE2 positive) are observed in the corneous layer of apteric epidermis but not in that of scutate scales, which contain mainly beta-keratin. Alpha-keratin bundles accumulate along the plasma membrane of transitional cells of apteric epidermis. In contrast to the situation in scutate scales, in the transitional layer and in the lowermost part of the corneous layer of apteric epidermis, filaggrin-like, loricrin-like, and transglutaminase immunoreactivities are present. The lack of isopeptide bond immunoreactivity suggests that undetectable isopeptide bonds are present in avian keratinocytes. Using immunogold ultrastructural immunocytochemistry a low but localized loricrin-like and, less, filaggrin-like labeling is seen over round-oval granules or vesicles among keratin bundles of upper spinosus and transitional keratinocytes of apteric epidermis. Filaggrin-and loricrin-labeling are absent in alpha-keratin bundles localized along the plasma membrane and in the corneous layer, formerly considered keratohyalin. Using ultrastructural autoradiography for tritiated histidine, occasional trace grains are seen among these alpha-keratin bundles. A different mechanism of redistribution of matrix and corneous cell envelope proteins probably operates in avian keratinocytes as compared to that of mammals. Keratin bundles are compacted around the lipid-core of apteric epidermis keratinocytes, which do not form complex chemico/mechanical-resistant corneous cell envelopes as in mammalian keratinocytes. These observations suggest that low amounts of matrix proteins are present among keratin bundles of avian keratinocytes and that keratohyalin granules are absent. PMID- 14755754 TI - Comparison of ThinPrep and conventional preparations in pancreatic fine-needle aspiration biopsy. AB - Use of ThinPrep preparation for fine-needle aspiration biopsy (FNA) is gaining popularity. However, there may be a difference in the morphology and the operating characteristics between ThinPrep and conventional methods. The objective of this study was to compare the accuracy of the two methods and to address the pitfalls of ThinPrep preparation in pancreatic FNA. A computer search identified 67 pancreatic FNAs with both conventional smears and ThinPrep preparation during a 19-mo period. These cases, obtained under endoscopic ultrasound-guidance, consisted of 47 malignant neoplasms (44 ductal carcinomas, two mucinous neoplasms, and one islet cell tumor) and 20 benign lesions. Direct smears were prepared first and the remaining material was then put into PreservCyt Solution for ThinPrep slides. All slides were reviewed and the cytologic diagnoses were correlated with histologic and clinical follow-up. Five conventional and 16 ThinPrep specimens were unsatisfactory due to insufficient cellularity. These cases were excluded from the analysis. Among the 62 cases evaluated by conventional preparation, 77% (34) were diagnosed as positive and 14% (seven) atypical/suspicious by conventional smears. For the 51 ThinPrep specimens, 58% (22) were interpreted as positive and 31% (12) atypical/suspicious. The sensitivity, specificity, and accuracy of diagnosing a malignancy were 77%, 100%, and 84% for conventional smears and 58%, 100%, and 67% for ThinPrep preparation, respectively. There were no false positives with either method. However, three benign lesions were interpreted as atypical/suspicious with ThinPrep preparation because of the presence of single atypical cells with distinct nucleoli. One of the two mucinous neoplasms was incorrectly diagnosed with ThinPrep preparation because of lack of mucin. The diagnostic accuracy of pancreatic FNA using ThinPrep is inferior to that of conventional smears. This may be partly due to the use of split sample technique resulting in scant cellularity in ThinPrep preparation and partly due to the differences in morphology between the two preparations. Therefore, the current morphologic criteria may need modification for ThinPrep preparation in pancreatic FNA. PMID- 14755755 TI - Gliosarcoma: cytopathologic characteristics on fine-needle aspiration (FNA) and intraoperative touch imprint. AB - Gliosarcoma (GS) is a rare subtype of glioblastoma multiforme (GBM), which shows a dimorphic population of glial and mesenchymal elements. The cytopathology of GS to our knowledge has not been previously described. Although prognostically insignificant within the group of GBM, an accurate recognition of this subtype may help to rule out other morphologically similar primary and metastatic central nervous system (CNS) neoplasms. Thirteen cases of histologically confirmed GS with concomitant touch imprints (TI) or prior fine-needle aspiration (FNA) were retrieved from the files of The Johns Hopkins Hospital (1985-2002). A comprehensive review of the clinico-radiologic, cytologic, and histologic material was undertaken to define the morphologic characteristics of GS. Material was obtained via computerized tomography (CT)-guided needle biopsy. Slides were stained with DiffQuik and/or hematoxylin and eosin (H and E) stains. Smears were highly cellular and showed a high-grade neoplasm with glial and mesenchymal elements. The latter component, however, predominated and showed a variety of phenotypic patterns, which included fibrosarcoma-like, rhabdoid type, osteoclastic giant cell type, undifferentiated type, and tumor with heterologous components (such as chondroid or osteoid tissue). A rich arborizing capillary network was evident, as were a high mitosis/karyorrhexis index and foci of necrosis. The glial component consisted of pleomorphic round to oval nuclei and numerous gemistocytes embedded in a fibrillary stroma. PMID- 14755756 TI - Extrapulmonary tuberculosis as a mimicker of neoplasia. AB - Despite the efforts for control and eradication of tuberculosis, new cases of the disease are diagnosed daily. The diagnosis of tuberculosis is easily made when the classical features of pulmonary necrotizing granulomatous inflammation are seen. However, extrapulmonary lesions may clinically and radiographically mimic a neoplastic process, and this may lead to misdiagnosis and delay in treatment. We studied 6 patients by aspiration biopsy, all recent immigrants and immunocompetent, who presented with weight loss and fatigue. Of these, 5 patients had a mass. One patient presented with a lytic lesion of bone. In all cases the clinical diagnosis was neoplasia. In all aspirates, the smears showed necrotic debris with neutrophils. No neoplastic cells or granulomas were seen. All cases were signed out descriptively with no specific diagnosis. A search for acid-fast organisms leading to the correct diagnosis of tuberculosis was prompted by clinical investigations that revealed pulmonary lesions, or by repeat aspiration biopsy, which showed granulomatous inflammation. Tuberculosis when present in atypical forms is still a challenging diagnosis. The finding of necrotic debris in a needle biopsy without the clinical signs of an abscess should prompt a search for acid-fast bacilli, since the correct diagnosis will eliminate a needless surgical procedure and will lead to timely and appropriate therapy. PMID- 14755757 TI - Superficial endometriosis of the cervix: A source of abnormal glandular cells on cervicovaginal smears. AB - Superficial endometriosis of the cervix, a benign process which may be associated with atypical glandular cells of undetermined significance (AGUS) on cervicovaginal (CV) smears, is becoming increasingly recognized on colposcopic examination. This study details the clinical, cytologic, and histology features of six cases of superficial cervical endometriosis. All CV smears featured atypical endocervical-like columnar cells in sheets and strips as well as cells with endometrial characteristics including solid cohesive, crowded, overlapping glandular groups, loss of cellular polarity, and a frequent ragged "feathered" edge appearance with protruding nuclei, occasional rosette formations, and endometrial stroma. Recognition of endometrial stroma in continuity with groups of cells with these features on CV smears may suggest the diagnosis of this benign condition. However, the cytologic features of endometriosis show sufficient overlap with those of precancerous and cancerous glandular lesions that many of these cases will continue to be diagnosed as "atypical glandular cells." PMID- 14755758 TI - Diagnostic utility of CD10 in differentiating hepatocellular carcinoma from metastatic carcinoma in fine-needle aspiration biopsy (FNAB) of the liver. AB - The differential diagnosis between hepatocellular carcinoma (HCC) and metastatic carcinoma, especially in moderate-poorly differentiated (MPD) HCC and poorly differentiated carcinoma, can be challenging in fine-needle aspiration biopsy (FNAB) of the liver. Recent studies demonstrate that canalicular staining for CD10 appears to be a highly specific marker for hepatocytic differentiation. The objective of this study was to test the utility of CD10 in differentiating HCC from metastatic carcinoma in FNAB of the liver. Formalin-fixed, paraffin-embedded cell blocks of 55 cases (22 HCC, 23 metastases, and 10 benign hepatic lesions) of FNAB of the liver were immunostained using monoclonal antibody against CD10, with microwave oven antigen retrieval, followed by a standard ABC method. Nineteen (86%) of 22 HCC cases were positive for CD10 with a canalicular staining pattern. Among them, 9 (82%) of 11 well-differentiated (WD) HCC and 10 (91%) of 11 MPD HCC were positive for CD10. Three (13%) of 23 metastatic carcinomas were positive for CD10, demonstrating a contrasting cytoplasmic and membranous staining pattern. The three positive cases were metastatic renal cell carcinoma (RCC), choriocarcinoma, and adenocarcinoma of the lung. All 10 cases of benign hepatic lesions showed positivity for CD10 with a canalicular and focal membranous staining pattern. In conclusion, CD10 appears to be a useful marker in discriminating between HCC and metastatic carcinoma when applied to FNAB of the liver. CD10 does not provide discrimination between WD HCC and benign hepatocytes. PMID- 14755759 TI - Differentiating nonhigh-grade duct carcinoma in situ from benign breast lesions. AB - This study was undertaken to determine the discriminating cytological features between nonhigh-grade duct carcinoma in situ (NHGDCIS) and benign breast lesions and to determine any histological characteristics which would influence the cytological categorization. Smears of 12 each of histologically confirmed NHGDCIS and benign breast lesions were reviewed with regard to cellularity, cell discohesion, nuclear atypia, crowding of cells, tubule formation, necrosis, and presence of bare atypical nuclei and regular bare bipolar nuclei, and statistically analyzed. Architectural pattern, presence of necrosis, and the size of the lesion assessed at histological examination were compared with the initial cytological categorization. NHGDCIS lesions showed more cell discohesion (P = 0.04), bare atypical nuclei (P = 0.05), necrosis (P = 0.03), and sparse bare bipolar nuclei (P = 0.02) than benign lesions. These differences were statistically significant. Cellularity (P = 0.8), nuclear atypia (P = 0.06), crowding of cells (P = 0.1), and tubule formation did not show a significant difference. Six (out of six lesions) with a solid architectural pattern and six (of seven) with necrosis could be cytologically categorized as suspicious or malignant. Size of the lesion did not influence this. We conclude that cell discohesion, bare atypical and bare bipolar nuclei, and necrosis are discriminating features between NHGDCIS and benign breast lesions and NHGDCIS lesions with a solid architectural pattern and necrosis are more likely to be satisfactorily categorized cytologically. PMID- 14755760 TI - Cutaneous alternariosis: cytohistological findings in a case diagnosed by fine needle aspiration biopsy. PMID- 14755761 TI - Factors affecting the adoption of new cytology technologies. PMID- 14755762 TI - FocalPoint slide classification algorithms show robust performance in classification of high-grade lesions on SurePath liquid-based cervical cytology slides. AB - The FocalPoint Primary Screening System (FPPS) operates by assigning scores to each slide relative to the probability that an abnormality is present. This information ranks each slide within five "quintiles" (1 = highest risk, 5 = lowest risk) of the "review" population, allowing examining cytologists to understand the risk inherent in each slide. Such information helps to make the primary and quality control rescreening processes most efficient. This study examines the efficiency of AutoPap scoring and, thus, stratification of high grade cases within a clinical trial setting. A total of 1,275 SurePath (TriPath, Burlington, NC) slides were screened on the FPPS. There were 124 high-grade cases in the set (32 HSIL, 5 AIS, and 87 cancers) as determined by cytologic truth adjudication. The efficiency of FPPS ranking was determined by analysis of the numbers of high-grade cases ranked into quintiles 1 (top 20%) and 1+2 (top 40%). FPPS places cases scored as "unsatisfactory" (HSIL, 3; Cancer, 18) into quintile 5 to ensure a manual review. These cases were excluded from the analysis. Overall, 58% of high-grade slides were classified as Q1 and 83% were classified as Q1+Q2. For HSIL, 66% were classified as Q1 and 90% as Q1+Q2. For Cancer, 59% were classified as Q1 and 84% as Q1+Q2. No high-grade slides were ranked in the lower "no review" population (slides that would receive no manual examination). The results confirm the robust performance of the FPPS classification algorithm. Far more high-grade slides are classified into the top two quintiles than would be expected by random chance alone (20% and 40%, respectively). These results validate the safety and effectiveness of this device on SurePath liquid-based slides. PMID- 14755763 TI - Aspiration cytology of focal xanthogranulomatous pyelonephritis: a diagnostic challenge. AB - A case of focal xanthogranulomatous pyelonephritis presenting as a circumscribed mass in a functioning kidney of a young male is described. Clinical, radiological, and cytological features closely mimicked renal cell carcinoma leading to radical nephrectomy. Peroperative findings and gross pathological features also corroborated preoperative diagnosis. The final diagnosis was a histologic surprise. It is an uncommon entity with cortical location, no pelvic communication, and progressively destroying the kidney. It should be considered in the differential diagnosis of any mass lesion at any age during preoperative clinicoradiological and cytological evaluation. A correct preoperative diagnosis can save unnecessary nephrectomy. PMID- 14755764 TI - Fine-needle aspiration of an adenoid cystic carcinoma of the larynx mimicking a thyroid mass. AB - Adenoid cystic carcinoma (ACC) is most often primary in the major and minor salivary glands but can also arise from the submucosal seromucinous glands of the larynx and trachea. We report a case of adenoid cystic carcinoma of the larynx that presented as a diffuse swelling in the thyroid area. Fine-needle aspiration (FNA) was consistent with a neoplastic process, which was difficult to classify further but was felt to be of thyroid origin. Subsequent gross and histopathologic examination showed the lesion to represent an ACC arising from the larynx. This case highlights the need to be aware of unusual lesions that may arise in the region of the thyroid. Knowledge of these non-thyroidal lesions that can clinically mimic a thyroid mass will help in making the correct cytologic diagnosis when these lesions are sampled by FNA. PMID- 14755765 TI - Fine-needle aspiration biopsy diagnosis of intrathoracic extramedullary hematopoiesis presenting as a posterior mediastinal tumor in a patient with sickle-cell disease: Case report. AB - Extramedullary hematopoiesis (EMH) is a rare cause of an intrathoracic mass. Fine needle aspiration biopsy (FNAB) has been only occasionally documented as a useful tool in diagnosing EMH tumor. We report a case of posterior mediastinal extramedullary hematopoietic mass in an 80-yr-old man with sickle-cell anemia. The mass was revealed incidentally on chest X-ray. The definitive diagnosis of this mass lesion was achieved by FNAB. The cytologic smears showed hematopoietic elements with erythroid hyperplasia. A correct cytologic diagnosis can thus help to avoid unnecessary surgical intervention, particularly in an asymptomatic patient. PMID- 14755766 TI - Mixed follicular-medullary thyroid carcinoma: a case report. AB - We report on a 35-yr-old woman presenting with a single thyroid nodule. Fine needle aspiration (FNA) of the nodule was reported to contain both follicular and parafollicular cells. The biphasic nature of the tumor was highlighted on immunohistochemical investigation of the cellblock. Positive staining for thyroglobulin was limited to the follicular structures and the dense areas stained positive for calcitonin. The serum calcitonin level was highly elevated. Surgery was recommended because of suspected malignancy. The patient underwent total thyroidectomy. The diagnosis of mixed medullary and follicular carcinoma of the thyroid was established by histological investigation using immunohistochemical staining for thyroglobulin, chromogranin, and calcitonin. The patient was well 1 yr after the operation. Repeated measurements of serum calcitonin levels were normal. Total body scan revealed no radioactive iodine uptake in the thyroid bed, bones, or lungs. PMID- 14755767 TI - Analysis of routine cytopathologic reports in 1,598 histologically verified benign breast lesions. AB - This retrospective study was designed to evaluate the accuracy of cytopathologic diagnosis and of correct classification of benign breast diseases. A total of 1,598 FNABs were identified to have met the study criteria; of these, 1,258 (78.7%) cases were cytologically benign, 88 (5.5%) suspicious, 3 (0.18%) false positive, and in 249 (15.6%) cases an inadequate sample was obtained. A specific diagnosis was made in 847/1,258 (67.3%) cases; the other 411 were diagnosed as benign NOS. Out of 847 specific FNABs diagnoses, 451 were fibroadenomas, 27 phyllodes tumors, 289 fibrocystic diseases, 4 proliferative fibrocystic diseases, 38 papillomas, 22 fat necrosis, 9 mastitis, 1 pseudolymphoma, 2 lipomas, 2 duct ecstasies, and 2 atheromas. In our study group the cytopathologic diagnosis of benign breast diseases excluding unsatisfactory aspirates was correct in 93%. Specific diagnosis was correct on average in 50% of cases, only in FA was its accuracy over 60%; in adequately sampled tumor, the predictive value of FA was 86.2%. PMID- 14755768 TI - Endometrial glands in ascites secondary to endometriosis. PMID- 14755769 TI - Yet more analogies in cytopathology. PMID- 14755770 TI - Cytological findings of carbon breast granuloma following stereotactic track localization. PMID- 14755771 TI - De novo breast cancer in patients with liver transplantation: University of Pittsburgh's experience and review of the literature. AB - De novo malignancies are one of the current problems in patients with organ transplantation. The incidence has been considered to be higher as a result of increases of oncogenic viruses in immunosuppressed organ recipients. Published reports have shown increased incidence of de novo tumors such as malignant lymphomas and cutaneous neoplasms but decreased incidence of breast cancer. A variety of factors affect de novo breast cancer development in organ recipients, including immunosuppression, viruses, and underlying disease. The aims of this review are to evaluate the incidence and management of patients with de novo breast cancer by giving the University of Pittsburgh's data, and to evaluate the incidence of de novo breast cancer in published reports in light of an age adjusted rate. According to age-adjusted rates presented by the National Cancer Institute's Surveillance, Epidemiology and End Results data, we found increased incidence rate of de novo breast cancer in the previously published series. The University of Pittsburgh's incidence rate of de novo breast cancer was determined in a fashion similar to that for the Surveillance, Epidemiology and End Results data. Eighty-three percent of all patients were diagnosed at early stages, and it appeared to take longer for de novo breast cancer to develop in patients treated with tacrolimus than in patients treated with cyclosporine. In conclusion, surgical treatment of breast cancer in liver recipients is the same as treatment of breast cancer in patients without transplantation. However, the effects of chemotherapy, radiotherapy, and/or tamoxifen remain unclear in transplanted patients and need to be evaluated in larger studies. PMID- 14755772 TI - Results of the first year of the new liver allocation plan. AB - Liver allocation policy in the U.S. was recently changed to a continuous disease severity scale with minimal weight given to time waiting in an effort to better prioritize deceased donor liver transplant candidates. We compared rates of waiting list registrations, removals, transplants, and deaths during the year prior to implementation of the new liver allocation policy (2/27/01-2/26/02, Era 1) with the first year's experience (2/27/02-2/26/03, Era 2) under this new policy. Rates were adjusted for 1,000 patient years on the waiting list and compared using z-tests. A 1-sided test was used to compare death rates; 2-sided tests were used to compare transplant rates. Overall and subgroup analyses were performed for demographic, geographic, and medical strata. In Era 2, we observed a 12% reduction in new liver transplant waiting list registrations, with the largest reductions seen in new registrants with low MELD/PELD scores. In Era 2, there was a 3.5% reduction in waiting list death rate (P =.076) and a 10.2% increase in cadaveric transplants (P <.001). The reduction in waiting list mortality and increase in transplantation rates were evenly distributed across all demographic and medical strata, with some variation across geographic variables. Early patient and graft survival after deceased donor liver transplantation remains unchanged. In conclusion, by eliminating the categorical waiting list prioritization system that emphasized time waiting, the new system has been associated with reduced registrations and improved transplantation rates without increased mortality rates for individual groups of waiting candidates or changes in early transplant survival rates. PMID- 14755773 TI - Periportal edema and necrosis as diagnostic histological features of early humoral rejection in ABO-incompatible liver transplantation. AB - Humoral rejection caused by antidonor blood group A/B antibodies is one of the most important obstacles for successful ABO-incompatible liver transplantation. However, no specific morphologic features of liver biopsies to distinguish humoral rejection from other conditions such as ischemia or sepsis have been satisfactorily documented. To histologically clarify the early changes in humoral rejection, we studied 41 cases of living donor ABO-incompatible liver transplantation whose allograft biopsies during the first episode of suspected acute rejection were available within the first postoperative month. Postoperative isohemagglutinin IgM titers were x64 or more in 21 patients (51%; high-titer group) and less than x64 in 20 cases (49%; low-titer group). In the high-titer group, elevation of postoperative titers x64 or more occurred within postoperative days 5.7 +/- 4.1 (range: 1-17). An increase in the incidence of cholangitis was observed in the high-titer group (90% vs. 30%, P <.0001), as well as poorer overall graft survival than in the low-titer group (38% vs. 70%, P <.05). Seven biopsies obtained from the high-titer group within 3 days after the onset of elevation of the antibody titers and one biopsy obtained at the peak of the antibody titers demonstrated periportal edema and necrosis, neither of which was found in the low-titer group. All grafts of these patients caused massive hepatocyte necrosis or severe biliary complications. In conclusion, a high morbidity rate of ABO-incompatible liver transplantation is associated with high postoperative levels of antibody titers. Periportal edema and necrosis observed during elevation of antibody titers can be regarded as histological indications of early changes in severe humoral rejection. PMID- 14755774 TI - Impaired regeneration of biliary cells in human chronic liver allograft rejection. Special emphasis on the role of the finest branches of the biliary tree. AB - Severe loss of bile ducts is a hallmark of chronic liver rejection. We hypothesize that loss of the finest branches of the biliary tree, including the intralobular segments, contributes to an impaired regenerative response of bile ducts in chronic rejection. The number and proliferative response of bile ducts, intraportal ductules, and extraportal biliary cells were studied in graft biopsies of 12 chronic-rejection patients. Twenty-two long-term-surviving patients who experienced acute rejection without chronic rejection served as controls. Reperfusion, 1-week, 1-month, and 1-year biopsies were studied. Monochlonal antibody Ki67 was applied to assess proliferation, and cytokeratin 7 antibody to identify bile ducts, intraportal ductules, and extraportal biliary cells. Progressive loss of bile ducts, intraportal ductules, and extraportal biliary cells was observed in the chronic-rejection group. In controls, all of these structures remained well preserved. Additionally, a significant increase of intraportal ductules was present at 1 week in controls, which was not the case in the chronic-rejection group. Proliferative activity of intraportal ductules and extraportal biliary cells was significantly increased at 1 week in controls. This proliferative activity was higher in the intraportal ductules of controls, compared with the chronic-rejection group. After 1 week, proliferative activity was virtually absent in both groups. In conclusion, our results showed that a deficient proliferative response of the finer branches of the biliary tree, including its intralobular segments, might contribute to bile-duct loss in chronic rejection. This finding supports the postulation that these structures represent a regenerative compartment of the biliary unit in the liver. PMID- 14755775 TI - Liver transplantation for hepatocellular carcinoma: the MELD impact. AB - The new allocation policy of the United Network of Organ Sharing (UNOS) based on the model for end-stage liver disease (MELD) gives candidates with stage T1 or stage T2 hepatocellular carcinoma (HCC) a priority MELD score beyond their degree of hepatic decompensation. The aim of this study was to determine the impact of the new allocation policy on HCC candidates before and after the institution of MELD. The UNOS database was reviewed for all HCC candidates listed between July 1999 and July 2002. The candidates were grouped by two time periods, based on the date of implementation of new allocation policy of February 27, 2002. Pre-MELD candidates were listed for deceased donor liver transplantation (DDLT) before February 27,2002, and post-MELD candidates were listed after February 27, 2002. Candidates were compared by incidence of DDLT, time to DDLT, and dropout rate from the waiting list because of clinical deterioration or death, and survival while waiting and after DDLT. Incidence rates calculated for pre-MELD and post MELD periods were expressed in person years. During the study, 2,074 HCC candidates were listed for DDLT in the UNOS database. The DDLT incidence rate was 0.439 transplant/person years pre-MELD and 1.454 transplant/person years post MELD (P < 0.001). The time to DDLT was 2.28 years pre-MELD and 0.69 years post MELD (P < 0.001). The 5-month dropout rate was 16.5% pre-MELD and 8.5% post-MELD (P < 0.001). The 5-month waiting-list survival was 90.3% pre-MELD and 95.7% post MELD (P < 0.001). The 5-month survival after DDLT was similar for both time periods. The new allocation policy has led to an increased incidence rate of DDLT in HCC candidates. Furthermore, the 5-month dropout rate has decreased significantly. In addition, 5-month survival while waiting has increased in the post-MELD period. Thus, the new MELD-based allocation policy has benefited HCC candidates. PMID- 14755777 TI - Liver allocation for HCC: a moving target. PMID- 14755776 TI - Impact of pretransplant diagnosis of hepatocellular carcinoma on cadveric liver allocation in the era of MELD. AB - The allocation system based on the Model for End-stage Liver Disease (MELD) has led to more patients diagnosed with hepatocellular carcinoma (HCC) being transplanted. We hypothesized that more patients misdiagnosed with HCC are also being transplanted, leading to inappropriate organ allocation. Therefore, we retrospectively analyzed all liver transplants at our center from July 14, 2000, to October 22, 2002 (N = 172; 129 pre-MELD, 43 post-MELD), comparing pretransplant HCC diagnosis to explant histology. Thirty patients met the United Network for Organ Sharing (UNOS) diagnostic criteria for pretransplant HCC diagnosis. There were 25 men (median age, 52.5 yr), and 80% had hepatitis C. The proportion of patients transplanted who had an HCC diagnosis increased from 12% (15/129) pre-MELD to 35% (15/43) post-MELD implementation (P < 0.01). Three of 15 (20%) transplanted pre-MELD and 5 of 15 (33%) transplanted post-MELD lacked HCC in the explant (P = 0.10). Of the three false-positives pre-MELD, one was Status 2B already, and two received living-donor livers. Of the 5 false-positives post MELD, three had score upgrades that led to early transplantation (13 to 29, 20 to 29, and 9 to 24) while two had MELD scores of 35 and 36 already. The percentage of organs that could have gone to patients with more advanced liver disease without HCC increased from 0% (0/129) pre-MELD to 7% (3/43) post-MELD (P < 0.01). Since the implementation of MELD, the proportion of patients transplanted who had an HCC diagnosis nearly tripled, and a small but significant proportion of organs are now going to patients misdiagnosed with HCC. More stringent HCC diagnostic criteria will be required to decrease the effect that misdiagnosis has on organ allocation. PMID- 14755779 TI - Hepatitis B virus genotype A and D and clinical outcomes of liver transplantation for HBV-related disease. AB - Hepatitis B virus (HBV) genotypes have been associated with specific patterns of disease and response to antiviral therapy. We investigated the effect of HBV genotype on HBV recurrence and mortality after liver transplantation (LT). Pretransplant sera of 45 hepatitis B surface antigen (HBsAg) positive adults were submitted for HBV genotyping by a reverse-phase hybridization line probe assay with genotype-specific probes. Data were correlated with clinical outcomes after transplantation. Genotype A (n =15), D (n = 13) and A/D (n = 12) accounted for 89% of all genotypes. Coinfection with two HBV genotypes was encountered in 14 (31.1%) patients. Eighteen patients (40 %) developed HBV recurrence at a median of 10 months posttransplant (range, 1-53) and 10 patients (22 %) died at a median of 24 months (range, 3-63). Genotype D patients were more likely to develop HBV recurrence or die compared with genotype A patients, although this did not reach statistical significance. Dual infection with genotype A/D resulted in mortality similar to that of genotype A but recurrence similar to that of genotype D. Active viral replication at time of transplantation was the only independent factor (P = 0.03) that predicted HBV recurrence. In conclusion, HBV genotype A and D did not have a significant impact on clinical outcomes of LT for HBV related liver disease in patients of European origin. These data do not support routine HBV genotyping in liver transplantation. PMID- 14755778 TI - A prospective cross-over study comparing the effect of mycophenolate versus azathioprine on allograft function and viral load in liver transplant recipients with recurrent chronic HCV infection. AB - It has been suggested that Mycophenolate Mofetil (MMF) may have an antiviral effect in patients with recurrent HCV infection post-liver transplantation. We conducted a prospective cross-over study in liver transplant recipients with recurrent chronic HCV infection to examine whether substitution of azathioprine (AZA) with MMF would reduce HCV load and improve allograft function. Thirteen long standing HCV liver transplant recipients receiving AZA were enrolled in a 9 month prospective cross-over study. In the initial 3 months lead-in period, baseline viral loads and biochemistry were recorded. Following this, MMF was substituted for AZA at a dose of 1 gm twice/day for a period of 3 months after which patients were switched back to AZA and observed for a further 3 months. Viral loads, biochemical allograft function, and adverse effects were closely monitored during the study period. Thirteen patients (12 males and 1 female) were enrolled. The mean age was 54 (+/-8) years and the mean time from transplantation was 68 (+/-35) months. Baseline mean viral load was 0.74 x 10(6)(+/-0.47 x 10(6)) messenger RNA (mRNA) copies/ml. By the end of the MMF treatment period, the mean viral load increased to a level of 1.64 x 10(6) (+/-1.3 x 10(6)) mRNA copies/ml (P = 0.026) compared to baseline. The increase in viral load however was not associated with an increase in ALT level. In a cohort of 13 HCV liver transplant recipients with recurrent chronic HCV infection, substitution of azathioprine with MMF did not lead to a decrease in viral load. PMID- 14755780 TI - Volume regeneration after right liver donation. AB - After right hepatectomy with the middle hepatic vein trunk for a graft, the venous outflow in segment IV is disturbed. There are limited data, however, regarding the effect of middle hepatic vein deprivation on liver regeneration or functional recovery. Living donors who underwent right hepatectomy with preservation of the middle hepatic vein (Group A, n = 58) and those deprived of the middle hepatic vein (Group B, n = 13) were reviewed. When the donor was under 50 years old and the remnant left liver was estimated to be more than 35% of the whole liver, right liver graft harvesting with the middle hepatic vein trunk was considered. Volume regeneration of segments I-III, segment IV, and overall liver volume was assessed at the third postoperative month using computed tomography. The regeneration rate of segment IV was significantly impaired in Group B donors compared with that in Group A donors (125% vs. 45%, P = 0.008). In contrast, the regeneration rate of segments I -III was significantly higher than that in Group A (208% vs. 263%, P = 0.004). There was no significant difference in the regeneration rate of the whole left liver or functional recovery between groups. Multivariate analysis revealed that the resection type (group) was a significant predictive factor for the regeneration rate of segments I-III and segment IV. When deprived of the middle hepatic vein, liver regeneration of segment IV was impaired but was compensated for by the regeneration of segments I-III. In conclusion, extended right hepatectomy can be safely performed with careful preoperative consideration using these criteria. PMID- 14755781 TI - Harvesting the middle hepatic vein with a right hepatectomy does not increase the risk for the donor. AB - The harvesting of the middle hepatic vein (MHV) with a right hepatectomy for living-donor liver transplantation allows an optimal venous drainage for the recipient but can also have adverse effects for the donor. This study compares morbidity, early liver function, and volume regeneration in 2 groups of donors who underwent right hepatectomy with (MHV+, n = 21) or without (MHV-, n = 20) MHV harvesting during 2 successive periods. The operative time was 401 +/- 60 minutes in the MHV+ group compared with 392 +/- 63 minutes in the MHV- group, and the transection time was 152 +/- 53 minutes in the MHV+ group compared with 131 +/- 30 minutes in the MHV- group (not significant). Blood loss in the MHV+ group was 773 +/- 343 mL compared with 613 +/- 361 mL in the MHV- group (not significant). The graft weight and remnant liver volume ratio were similar in the MHV+ and MHV- groups (763 +/- 200 gm vs. 832 +/- 156 gm and 42% +/- 9.5% vs. 43% +/- 8.3%, respectively). Postoperative biologic liver function tests showed that prothrombin time (PT) ratio on postoperative days 1 and 3 were significantly lower in the MHV+ group compared with the MHV- group (53% vs. 65% and 63% vs. 72%, respectively, P <.05). There were no differences in postoperative alanine aminotransferase and aspartate aminotransferase peak levels between the MHV+ and MHV- groups (319 +/- 198 IU /L vs. 310 +/- 110 IU /L and 317 +/- 226 IU /L vs. 296 +/- 125 IU /L, respectively). Bilirubin maximal blood level was similar in the 2 groups (32 +/- 17 micromol/L in the MHV+ group vs. 43 +/- 16 micromol/L in the MHV- group, P <.05). No donor died. The overall morbidity was lower in the MHV+ group compared with the MHV- group (36% vs. 55%; P >.05, not significant). The donor's remnant liver volume regeneration, evaluated by computed tomography (CT) volumetric study on day 7, was similar in the 2 groups (97% +/- 29% in the MHV+ group and 103% +/- 39% in the MHV- group, P >.05). The results of this comparative study show that right hepatectomy with the MHV neither affects morbidity nor impairs early liver function and regeneration in donors. PMID- 14755782 TI - Incomplete improvement of visuo-motor deficits in patients with minimal hepatic encephalopathy after liver transplantation. AB - Previous studies have suggested reversibility of minimal hepatic encephalopathy in patients with liver cirrhosis after liver transplantation (LT), however, this topic is controversially discussed. We investigated this issue in a prospective study on liver cirrhotic patients listed for LT. Patients were investigated before and after liver transplantation (on average 21 months later) using a neuropsychological test battery which measured visuo-constructive and visuo-motor ability, verbal fluency, and memory function. To assess visuo-motor and visuo constructive functions, we performed 4 tests: Rey Complex Figure Test copy, trail making tests A and B, and digital symbol test. The average percentile score of the tests, arbitrarily named the visuo-motor and visuo-constructive performance score (VMCP), was calculated. After LT, the patients did not demonstrate a significant increase of VMCP (P =.29) and additionally showed significantly lower VMCP score (P =.041) compared to control group. Analysis of individual responses showed that only 7 of 14 patients improved their VMCP values after LT. These data indicate that the cirrhosis-associated visuo-motor deficits subside or disappear only in some of the patients after LT, whereas a significant number of patients show no improvement of the visuo-motor and visuo-constructive function. We concluded that monitoring of cognitive and visuo-motor functioning is important for the post-transplant rehabilitation of patients with liver cirrhosis. PMID- 14755783 TI - Is minimal hepatic encephalopathy completely reversible following liver transplantation? PMID- 14755784 TI - Liver transplantation and health-related quality of life: scoring differences between men and women. AB - Orthotopic liver transplantation (OLT) is the treatment of choice for end-stage liver disease of various etiologies. Its use, however, remains limited due to the scarcity of donor organs. Measures to assess health-related quality of life (HRQOL) are increasingly being implemented to examine the efficacy of medical therapies involving scarce resources. HRQOL was assessed and compared between 88 male and 61 female patients before and after liver transplantation. Data were gathered from subjects having completed a questionnaire pre-OLT, and again at 1 year and 2 years post-OLT. This questionnaire, developed specifically for OLT patients, contains at its core questions derived from several well-established instruments measuring health status and HRQOL. Male OLT recipients reported a higher degree of overall HRQOL than that reported by female OLT recipients, both before and after OLT. When controlling for disparity in education between the sexes, findings revealed that among the lesser educated (< or =12 years), men and women scored similarly, while among the more educated (>12 years), men scored higher than women. Employment findings revealed a higher percentage of men working before transplant and at 1-year post-OLT when compared with women. At 2 years post-OLT, men and women exhibited similar employment rates. Male OLT recipients report a higher level of overall HRQOL than that reported by female OLT recipients, both before and after liver transplantation. Education appears to significantly affect HRQOL and may account for, at least in part, differences in reported HRQOL between male and female OLT recipients. PMID- 14755785 TI - A matched comparison study of medical and psychiatric complications and anesthesia and analgesia requirements in methadone-maintained liver transplant recipients. AB - Approximately 85% of patients receiving methadone maintenance therapy (MMT) for opiate dependence in the United States are infected with hepatitis C virus (HCV). MMT is significantly underrepresented in most liver transplant programs, but the number of patients receiving MMT is increasing and few data are available to guide treatment. We evaluated MMT in our program (27 pretransplant and 10 posttransplant cases) for medical and psychiatric complications and anesthesia and analgesia requirements. After transplant, 10 patients receiving MMT were compared with a matched control group of 19 patients who were not receiving MMT and not dependent on opiates. Fewer patients receiving MMT retained a spot on the transplant waiting list (65%) than patients not receiving MMT (80%); 30% of patients receiving MMT pretransplant used heroin, cocaine, or marijuana, and more than 25% were lost to follow-up. Liver disease according to mean Child-Turcotte Pugh (CTP) score and transplant waiting times was similar between the 2 groups. Patients receiving MMT required significantly more intraoperative anesthesia and postoperative analgesia (mean fentanyl 3,175 microg/d, SD = 2,832; intravenous morphine 67.86 mg/d, SD = 38.84, respectively) compared with patients not receiving MMT (mean fentanyl 1,324 microg/d, SD = 1,122; intravenous morphine 12.17 mg/d, SD = 10.24, respectively). More patients receiving MMT had severe recurrent HCV infection (60%) and worse survival (60%) versus patients not receiving MMT (21% and 78.9%, respectively). Follow-up times did not differ between groups (MMT: mean 4.19 years, median 1.15 years, SD = 7.6; non-MMT: mean 2.68 years, median 2.19 years, SD = 1.73). Finally, patients receiving MMT required an average methadone dose increase of 60% from pretransplant to posttransplant. Postoperative analgesia guidelines are described. Posttransplant, 20% of patients receiving MMT used alcohol or illicit drugs. Data do not support withholding the provision of liver transplantation to patients receiving MMT, but larger, well-controlled studies are warranted. PMID- 14755786 TI - Treatment of refractory cholestatic pruritus after liver transplantation with albumin dialysis. AB - Albumin dialysis has been shown to improve the outcome in patients with cholestatic liver failure caused by chronic liver disease. This study reports 7 liver transplant recipients who were treated with albumin dialysis for intractable pruritus of different origin (ductopenic graft rejection, non anastomotic strictures, and recurrence of hepatitis C). Treatment with histamine (H1) blockers, opioid antagonists, and cholestyramine had not been effective. The Molecular Adsorbent Recirculating System (MARS; Teraklin, Rostock, Germany) was used for albumin dialysis. All patients presented with numerous scratch marks, 6 of whom had a pronounced icterus. Six patients (86%) responded to 3 consecutive treatments with significant reduction of pruritus. The mean pruritus score, which was quantified by a visual analog scale (VAS), decreased from 9.7 +/- 0.5 to 3.7 +/- 0.8 (SD). The mean duration of 1 treatment was 15.6 hours. The procedure was well tolerated by all patients. The mean total serum bilirubin in patients who responded to therapy declined from 19.11 +/- 16.96 mg/dL (SD) before MARS therapy to 9.24 +/- 3.52 mg/dL after treatment. The mean serum concentration of 3 alpha hydroxy bile acids decreased from 192.67 +/- 58.12 micromol/L (SD) to 42.33 +/- 31.58 micromol/L (SD). Follow-up in 3 cases showed sustained improvement of pruritus lasting for more than 3 months. In 3 patients, however, pruritus relapsed. One patient, who showed severe pruritus, without relevant elevation of serum bile acids before treatment, did not respond to albumin dialysis. Our data indicate that MARS is an effective therapeutic option for patients with intractable cholestatic pruritus. PMID- 14755787 TI - Native liver xanthogranulomatous cholangiopathy in primary sclerosing cholangitis: impact on posttransplant outcome. AB - A retrospective analysis of 51 primary sclerosing cholangitis (PSC) patients who underwent liver transplant (LT) identified 16 with xanthogranulomatous cholangiopathy (XGC) at the native liver hilum. Pre-LT clinical and laboratory data and post-LT course and outcome of patients with XGC were compared with the 35 PSC patients without XGC. The XGC and non-XGC groups were similar with respect to age and laboratory data at the time of LT. Pre-LT cholecystectomy was performed in 44% versus 26% and biliary bypass procedure in 38% versus 26% of patients with and without XGC, respectively (P = NS). Peri-operative complications resulted in six (38%) deaths or retransplantation within 60 days of LT in the XGC group compared with 4 (11%) in the non-XGC group (P =.05). Patient survival at 60 and 100 days post-LT was better in the non-XGC group (P =.01). The causes of death or retransplantation within 60 days post-LT in the patients with XGC included primary nongraft function (1), uncontrolled bleeding (3), and sepsis (2), while in the non-XGC group these were uncontrolled bleeding (2), sepsis (1), and primary nongraft function (1). Mean graft survival +/- SD was 1,081 +/- 1,584 days in patients with XGC versus 2,149 +/- 1,679 days in patients without XGC. The presence of XGC in the native liver hilum of PSC patients undergoing LT was associated with a higher rate of early post-LT mortality or retransplantation. In conclusion, no pre-LT clinical features or laboratory tests were identified that predicted the presence of XGC in PSC patients. PMID- 14755788 TI - Quantitative assessment of fibrinogen cross-linking by epsilon aminocaproic acid in patients with end-stage liver disease. AB - Analysis of the effectiveness of antifibrinolytic therapy for liver transplant recipients is hampered by lack of quantitative assays for assessing drug effects. We adapted chemical engineering tools used in polymerization studies to quantify fibrinogen cross-linking by plasma from liver transplant patients obtained before and after epsilon aminocaproic acid (EACA) therapy. A target fluorescein isothiocyanate-fibrinogen (FITC-fibrinogen) molecule was constructed; it fluoresces in a quantifiable pattern when in solution, and undergoes cross linking in the presence of plasmin inhibitors. Cross-linking quenches the fluorescent signal, and the quenching is a quantifiable endpoint. Thus fluorescence from this reporter molecule can be used to assess functional improvement in fibrinogen cross-linking as a result of antifibrinolytic therapies, and it is sensitive to picomolar amounts of plasmin inhibitors and activators. Cross-linking of FITC-fibrinogen by patient plasma, before and after EACA therapy, was assessed using fluorescence spectrometry. Fluorescence patterns from FITC-fibrinogen indicated no significant cross-linking of the target fibrinogen as a consequence of EACA in posttreatment plasma. When the fibrinogen FITC target was assayed without plasma in the presence of EACA at concentrations that bracket therapeutic levels (100 and 400 microg/ml), significant fluorescence quenching (target FITC-fibrinogen cross-linking) was achieved. These results suggest that fibrinogen-FITC fluorescence is sensitive enough to detect EACA activity in clinically relevant ranges, but that EACA given in usual doses is insufficient to promote fibrinogen cross-linking in patients with end-stage liver disease. PMID- 14755789 TI - Hepatic arterial anatomy for right liver procurement from living donors. AB - Living donor liver transplantation (LDLT) using right liver grafts is now widely performed. Anatomic classifications of the hepatic artery for right liver procurement, however, are limited. In this study, celiac and mesenteric angiograms of 223 consecutive living donors in a single institution were evaluated. Details of the arterial anastomosis and results were reviewed in 72 patients who underwent primary LDLT using right liver grafts. There was a 6% incidence of hepatic arterial bifurcations that might provide multiple orifices in a right liver graft. Only one right liver graft (1%) had multiple arterial orifices. Single arterial anastomosis without interposition was possible in all patients with right liver grafts and none of them were complicated with hepatic arterial thrombosis. Single arterial anastomosis, therefore, has a high probability of success in right liver graft implantation. PMID- 14755790 TI - Hepatic arterial anatomy for right liver procurement from living donors. PMID- 14755791 TI - Pseudohypocalcemia after magnetic resonance imaging with gadolinium in patients with cirrhosis. AB - Hypocalcemia in patients with cirrhosis may be due to a number of causes. We noted a relationship between injection with gadodiamide, a particular gadolinium chelate, during magnetic resonance imaging of the liver and the development of a falsely low serum total calcium level in a patient with cirrhosis. A cross reference and retrospective chart review identified 10 additional patients in whom this phenomenon was noted. We describe the temporal relationship and clinical characteristics of these patients. Pseudohypocalcemia following magnetic resonance imaging with gadodiamide contrast should be considered in the differential diagnosis of hypocalcemia in patients with cirrhosis. PMID- 14755792 TI - Hepatectomy of living donors with a left-sided gallbladder and multiple combined anomalies for adult-to-adult living donor liver transplantation. AB - The left-sided gallbladder is very rare, but it is often accompanied by multiple anomalies of the liver, by which living donor hepatectomy cannot be feasible or becomes difficult. We have experienced 3 donors with a left-sided gallbladder out of 642 living donors. The first case was a male donor showing bifurcating portal anomaly with intrahepatic right portal vein confluence and extremely low bifurcation of the bile ducts. The right lobe was retrieved and implanted to his father. The second case was a male donor revealing trifurcating portal anomaly with separate right posterior portal branch and replacing right posterior hepatic artery. The right posterior segment graft was retrieved and implanted to his uncle. The third case was a male volunteer in whom the anterior portion of the medial segment was fed by an aberrant branch of the right anterior segment glisson. The small left lobe was retrieved and implanted simultaneously with another living donor's left lobe graft in the form of a dual living donor liver transplantation. There was no donor morbidity or recipient complication. Although there is a high possibility of diverse liver anomalies in living donors with a left-sided gallbladder, complete preoperative evaluation and mapping of the multiple anatomical variations may make certain types of living donor hepatectomy feasible. PMID- 14755793 TI - Laparoscopic iatrogeny of the hepatic hilum as an indication for liver transplantation. AB - The introduction of biliary laparoscopic surgery led to an increase in the incidence of liver hilum injuries. These types of lesions are very serious, because they can lead to secondary biliary cirrhosis or fulminant hepatic failure and the need for liver transplantation (LT). We present three cases of liver hilum injuries, which were treated with LT; one case was due to severe and persistent cholangitis, and two cases were due to fulminant hepatic failure. The world literature is also reviewed, and published cases of iatrogenic lesions of the liver hilum caused by laparoscopic surgery and requiring LT are presented. These iatrogenic lesions of the hepatic hilum are complex and technically demanding, due to their high morbidity and mortality and even the need for LT. In conclusion, these lesions must be always managed in centers with experience in hepatobiliary surgery. PMID- 14755794 TI - Split-liver transplants for two adult recipients: technique of preservation of the vena cava with the right lobe graft. PMID- 14755795 TI - Syndromic incidence of ovarian cancer after liver transplantation: is breast cancer an antecedent risk? PMID- 14755796 TI - Serum phosphate is not a reliable early predictor of outcome in paracetamol induced hepatotoxicity. PMID- 14755797 TI - TIPSS as therapeutic modality for umbilical hernia in patients with advanced liver disease. PMID- 14755799 TI - Analysis of RING finger genes required for embryogenesis in C. elegans. AB - The RING finger motif exists in E3 ligases of the ubiquitination pathway. These ubiquitin ligases bind to target proteins, leading to their modification by covalent addition of ubiquitin peptides. Current databases contain hundreds of proteins with RING finger motifs. This study investigates the role of RING finger genes in embryogenesis of the nematode, Caenorhabditis elegans. We expand the previous list of RING finger-containing genes and show that there are 103 RING finger-containing genes in the C. elegans genome. DNA microarrays of these 103 genes were probed with various RNA samples to identify 16 RING finger genes whose expression is enriched in the germline. RNA interference (RNAi) analysis was then used to determine the developmental role of these genes. One RING finger gene, C32D5.10, showed a dramatic larval arrest upon RNAi. Three RING finger genes exhibited embryonic lethality after RNAi. These three genes include par-2, and two small RING finger proteins: F35G12.9 (an ortholog of APC11) and ZK287.5 (an ortholog of rbx1). Embryos from RNAi of the APC11 and rbx1 orthologs were arrested in the cell cycle, confirming the role of these particular RING finger proteins in regulation of the cell cycle. genesis 38:1-12, 2004. PMID- 14755800 TI - Gastrulation events in the prestreak pig embryo: ultrastructure and cell markers. AB - The epithelial versus mesenchymal phenotypes of embryonic ectoderm and mesoderm cells of the prestreak stage pig embryos were examined by electron microscopy and molecular marker analysis. During this period the embryonic disc remained flat or slightly convex while becoming oval or pyriform in shape. Mesenchyme cells expressing vimentin were present between the embryonic disc and the underlying visceral endoderm before a primitive streak (or groove) was apparent. The migration of mesenchyme appeared to occur in lateral and posterior directions from a mass of quiescent cells located in the pointed end of the pyriform embryonic disc that expressed Brachyury; these cells are proposed to be the precursors of the primitive streak and/or form the equivalent of the mouse early gastrula organizer (EGO). Cells with the TEC-1 (or SSEA-1) epitope, the marker most frequently used to characterize pluripotent cells, were initially distributed randomly in the embryonic ectoderm and then were found to localize in an anterior crescent which may contain the precursor cells of ectoderm and neurectoderm. As mitotic figures were found only in the anterior crescent, it is proposed that at least some of these proliferating cells migrate toward the EGO. While cytokeratins were barely detectable in the embryonic ectoderm cells, vimentin expression was supposed to be associated with the migratory capacity of these cells. These findings indicate that the early step of gastrulation, migration of extraembryonic mesoderm, occurs at a prestreak stage during which the embryonic disc becomes polarized. genesis 38:13-25, 2004. PMID- 14755801 TI - Zebrafish intestinal fatty acid binding protein (I-FABP) gene promoter drives gut specific expression in stable transgenic fish. AB - Mammalian intestinal fatty acid-binding protein (I-FABP) is a small cytosolic protein and is thought to play a crucial role of intracellular fatty acid trafficking and metabolism in gut. To establish an in vivo system for investigating its tissue-specific regulation during zebrafish intestinal development, we isolated 5'-flanking sequences of the zebrafish L-FABP gene and used a transgenic strategy to generate gut-specific transgenic zebrafish with green/red fluorescent intestine. The 4.5-kb 5'-flanking sequence of zebrafish I FABP gene was sufficient to direct fluorescent expression in intestinal tube, first observed in 3 dpf embryos and then continuously to the adult stage. This pattern of transgenic expression is consistent with the expression pattern of the endogenous gene. In all five transgenic lines 45-52% of the F2 inheritance rates were consistent with the ratio of Mendelian segregation. These fish can also provide a valuable resource of labeled adult intestinal cells for in vivo or in vitro studies. Finally, it is possible to establish an in vivo system using these fish for screening genes required for gut development. genesis 38:26-31, 2004. PMID- 14755802 TI - Histone deacetylase activity is required for embryonic stem cell differentiation. AB - Mammalian development requires commitment of cells to restricted lineages, which requires epigenetic regulation of chromatin structure. Epigenetic modifications were examined during in vitro differentiation of murine embryonic stem (ES) cells. Global histone acetylation, a euchromatin marker, declines dramatically within 1 day of differentiation induction and partially rebounds by day 2. Histone H3-Lys9 methylation, a heterochromatin marker, increases during in vitro differentiation. Conversely, the euchromatin marker H3-Lys4 methylation transiently decreases, then increases to undifferentiated levels by day 4, and decreases by day 6. Global cytosine methylation, another heterochromatin marker, increases slightly during ES cell differentiation. Chromatin structure of the Oct4 and Brachyury gene promoters is modulated in concert with their pattern of expression during ES cell differentiation. Importantly, prevention of global histone deacetylation by treatment with trichostatin A prevents ES cell differentiation. Hence, ES cells undergo functionally important global and gene specific remodeling of chromatin structure during in vitro differentiation. genesis 38:32-38, 2004. PMID- 14755803 TI - General method for the modification of different BAC types and the rapid generation of BAC transgenic mice. AB - Most genome projects have relied on the sequencing of bacterial artificial chromosomes (BACs), which encompass 100-300 kb of genomic DNA. As a consequence, several thousand BAC clones are now mapped to the human and mouse genome. It is therefore possible to identify in silico a BAC clone that carries a particular gene and obtain it commercially. Given the large size of BACs, most if not all regulatory sequences of a gene are present and can be used to direct faithful and tissue-specific expression of heterologous genes in vitro in cell cultures and in vivo in BAC-transgenic mice. We describe here an optimized and comprehensive protocol to select, modify, and purify BACs in order to generate BAC-transgenic mice. Importantly, this protocol includes a method to generate, within 2 days, complex plasmid cassettes required to modify BACs, and to efficiently modify different types of BACs selected from the two major BAC libraries available. Altogether, using a combination of genomic database analysis, overlap PCR cloning, and BAC recombination in bacteria, our approach allows for the rapid and reliable generation of "pseudo knockin" mice. genesis 38:39-50, 2004. PMID- 14755804 TI - Ethical issues for invasive cardiologists: Society for cardiovascular angiography and interventions. AB - In view of the major impact of medical economic forces, rapidly changing technology, and other pressures on invasive cardiologists, the Society for Cardiovascular Angiography and Interventions determined that a statement of the ethical issues confronting the modern invasive cardiologist was needed. The various conflicts presented to the cardiologist in his or her roles as practicing clinician, administrator of the catheterization laboratory, educator, or clinical researcher were reviewed. In all instances, the major concern was determined to be the welfare of the patient no matter how forceful the pressures from various outside force or concerns for personal advancement might be. PMID- 14755805 TI - Reliable anticoagulation with enoxaparin in patients undergoing percutaneous coronary intervention: The pharmacokinetics of enoxaparin in PCI (PEPCI) study. AB - The objective of this study was to evaluate the pharmacokinetic response to intravenous (IV) enoxaparin given 8-12 hr after subcutaneous (SC) dosing in patients undergoing percutaneous coronary intervention (PCI). Fifty-five patients received SC enoxaparin (1 mg/kg every 12 hr) followed by an IV bolus (0.3 mg/kg) 8-12 hr after the last SC dose, at the start of PCI or during catheterization. Anti-Xa levels were within the target range in 98% of patients 2-8 hr after the last SC dose, in 96% of patients following the IV bolus, and in 91% of patients for a further 2 hr. Subcutaneous enoxaparin (1 mg/kg every 12 hr) provides sufficient anti-Xa levels for PCI 2-8 hr after the last dose. An additional 0.3 mg/kg enoxaparin dose given IV 8-12 hr after the last SC dose reliably maintains anti-Xa levels within the target for at least 2 additional hr. PMID- 14755806 TI - Seamless anticoagulation therapy utilizing enoxaparin for acute coronary syndromes: Measure or not, here it comes! PMID- 14755807 TI - Intracoronary brachytherapy, a promising treatment option for diabetic patients: Results from a European multicenter registry (RENO). AB - Despite advances in the interventional treatment of coronary disease, diabetics still have double the case fatality rate as nondiabetics. The purpose of this analysis from the Radiation in Europe With Novoste (RENO) registry was to assess the clinical and angiographic 6-month outcome of diabetic patients in comparison to nondiabetic patients after localized beta-radiation. A total of 1,098 patients (83.8% with in-stent restenosis) treated with the Novoste Beta-Cath system in Europe were enrolled in the RENO registry. Diabetes was, irrespective of the type of lesion treated, no significant risk factor for major adverse cardiac events or target vessel revascularization. Individuals with diabetes (n = 256) and without diabetes (n = 833) displayed no significant differences concerning clinical or angiographic endpoints. Vascular brachytherapy appears to be the first technique to even out the increased risk of diabetic patients undergoing percutaneous coronary interventions in the routine clinical setting. Thus, intracoronary brachytherapy represents a promising treatment option for diabetic patients. PMID- 14755808 TI - Safety of a high bolus dose of tirofiban in patients undergoing coronary stent placement. AB - To overcome the suboptimal platelet inhibition induced by tirofiban in the first hour after a percutaneous coronary intervention, a new regimen of 25 microg/kg bolus followed by an 18-hr infusion of 0.15 microg/kg/min has been proposed. The aim of this study was to compare the effects of this high bolus dose of tirofiban with those of abciximab on bleeding risk and 30-day clinical outcome in patients undergoing coronary stenting. We compared two cohorts of patients who underwent coronary stent placement between January 2000 and December 2002. In the first cohort, the only available IIb/IIIa receptor inhibitor was abciximab, which was given to 280 (34.9%) out of 802 stented patients; in the second cohort, tirofiban was administered to 274 (38.3%) out of 716 treated patients. The primary endpoints were the proportion of patients with major bleeding and the rate of site access complications; the 30-day incidence of major adverse cardiac events (MACE) was also assessed. After the procedure, the patients were given ticlopidine for 4 weeks and aspirin indefinitely. Major bleeding episodes were observed in four patients receiving abciximab and in none receiving tirofiban (1.4% vs. 0%; P = 0.12); the rates of site access complications were similar (3.6% vs. 3.3%; P = 0.96). The 30-day incidence of MACE was 7.1% in the abciximab group and 5.8% in the tirofiban group (P = 0.65). In patients undergoing coronary stenting, the high bolus dose of tirofiban is safe and not associated with an increased risk of major bleeding or site access complications in comparison with abciximab. PMID- 14755809 TI - GP IIb/IIIa inhibition with eptifibatide lowers levels of soluble CD40L and RANTES after percutaneous coronary intervention. AB - Platelets are the most abundant blood source of CD40L, a proinflammatory and prothrombotic costimulatory molecule implicated in atherosclerosis. Agonist stimulation results in the secretion of a soluble form of CD40L (sCD40L) and GP IIb/IIIa receptor inhibition blocks secretion of sCD40L in vitro. However, the effect of GP IIb/IIIa inhibition on sCD40L levels in humans is unknown. Plasma sCD40L and inflammatory markers were measured (t = 0, 0.5, 2, and 24 hr post-PCI) in a cohort of patients receiving abciximab (n = 15), eptifibatide (n = 15), or no GP IIb/IIIa inhibitor (n = 15). PCI in the absence of GP IIb/IIIa inhibitor was associated with a small but measurable rise in sCD40L and the platelet derived chemokine RANTES. In contrast, eptifibatide significantly lowered baseline sCD40L (P = 0.018) and RANTES (P = 0.006) levels. This effect was not observed with abciximab. GP IIb/IIIa inhibition with eptifibatide lowers levels of sCD40L and RANTES post-stenting, possibly conferring anti-inflammatory as well as antithrombotic effects. PMID- 14755810 TI - Intracoronary stenting and angiographic results: Restenosis after direct stenting versus stenting with predilation in patients with symptomatic coronary artery disease (ISAR-DIRECT trial). AB - The objective of this randomized study was to assess whether direct stenting leads to less restenosis than does conventional stenting (CS) with predilation in clinical practice. We included 910 patients who were randomly assigned to undergo either direct stenting (DS; n = 456) or CS (n = 454). No significant difference was observed in the incidence of angiographic restenosis (primary endpoint): 23.6% for DS and 21.0% for CS (P = 0.41; relative risk = 1.1; 95% CI = 0.8-1.5). The incidence of target vessel revascularization was 17.3% among DS and 14.8% among CS patients (P = 0.29; relative risk = 1.2; 95% CI = 0.8-1.6). The combined incidence of death or myocardial infarction at one year was 9.0% in the DS group and 7.0% in the CS group (P = 0.28). In conclusion, direct stenting is not associated with any reduction of thrombotic and restenotic complications as compared to the conventional stenting. PMID- 14755811 TI - Transcatheter closure of postinfarction ventricular septal defects using the new Amplatzer muscular VSD occluder: Results of a U.S. Registry. AB - The objective of this study was to assess the immediate and mid-term results of transcatheter closure of postinfarct muscular ventricular septal defects (VSDs) using the new Amplatzer postinfarct muscular VSD device (PIMVSD). Ventricular septal rupture occurs in 0.2% of myocardial infarcts and remains associated with very high morbidity and mortality. Data were prospectively collected for 18 patients who underwent attempted device closure of postinfarction VSDs between 2000 and 2003. Five patients underwent the closure in the acute phase (within 6 days from the infarct); the remaining patients underwent closure on day 14-95 after the diagnosis of the infarct. Outcome parameters included procedural success, evidence of residual shunts on echocardiography, and occurrence of procedure-related complications. The procedure was successful in deploying a device across the VSD in 16 of 18 patients. The 30-day mortality was 28%. Eleven patients are still alive and have been followed up for a median of 332 days. Two patients required a second procedure to close a residual VSD. At the most recent outpatient follow-up, the VSD was completely closed in two patients, six patients had a trivial or small residual shunt, and two patients had a moderate residual shunt. We conclude that percutaneous device closure of postinfarction VSDs using the Amplatzer PIMVSD occluder appears to be safe and effective. Further trials are required to assess long-term efficacy and compare the results with those of surgical closure. PMID- 14755812 TI - Rheolytic thrombectomy with distal filter embolic protection as adjunctive therapies to high-risk saphenous vein graft intervention. AB - Percutaneous intervention of saphenous vein graft (SVG), especially those with heavy atherothrombotic load, presents high risk for distal embolization and no reflow. Using the distal filters alone may occasionally be disadvantageous because of the large debris burden and the inability to assess the underlying culprit lesions and vessel size accurately. We present a case of intervention of an occluded SVG using a combination of rheolytic thrombectomy and distal filter embolic protection as a pretreatment before stenting. This strategy has the potential to reduce further the risk of no-reflow and to provide visualization for proper assessment of the underlying anatomy especially in clot-laden vessels. PMID- 14755813 TI - Kissing sirolimus-eluting stents for the treatment of left main coronary artery stenosis. AB - We present a case of kissing drug-eluting stent deployment in the left main coronary artery in a 43-year-old male with failed previous bypass grafting. We discuss the technique used for stent deployment and the rationale for using this technique in the drug-eluting stent era. PMID- 14755814 TI - Ulnar artery catheterization with occlusion of corresponding radial artery. AB - The transradial approach to coronary angiography has become a popular technique. Because of potential advantages, the transulnar approach has also recently been described. We report a successful case of transulnar catheterization with documented occlusion of the radial artery and normal inverse Allen test in a patient with limited vascular access. PMID- 14755815 TI - Coronary artery pseudoaneurysm after balloon angioplasty and intracoronary beta radiation for in-stent restenosis. AB - Intracoronary brachytherapy is an effective method for treating in-stent restenosis. We report a case of coronary artery pseudoaneurysm after balloon angioplasty and intracoronary beta-radiation. The pseudoaneurysm was treated successfully with implantation of two coronary stent grafts. PMID- 14755816 TI - Long-term follow-up of brachytherapy for treatment of allograft in-stent restenosis. AB - The experience of brachytherapy in the treatment of in-stent restenosis of allograft arteries is limited. We present two cases of in-stent restenosis treated with brachytherapy with favorable angiographic follow-up at 10 months in one patient and at 17 months in the other. PMID- 14755817 TI - Relationship between pattern of slow-reflow and tissue level of perfusion after coronary stenting: A single-patient report. AB - Ths study reports different mechanisms (i.e., microvascular spasm and discrete microinfarction) that may underlie different pattern of slow-reflow after elective coronary stenting on the basis of the different tissue level of perfusion achieved, assessed by blush grade and MRI. PMID- 14755818 TI - New strategies in the percutaneous management of coronary artery fistulae: A case report. AB - Coronary artery fistulae are rare anomalies often treated percutaneously. New techniques aiding in the percutaneous closure of a giant coronary artery, not previously described, are presented in a case report. These techniques include the use of intravascular ultrasound imaging and the use of electrolytically detachable platinum coils. PMID- 14755819 TI - TOR of the cell cycle: Are there important implications for diabetics in the era of the drug-eluting stent? PMID- 14755820 TI - Transcatheter closure of double atrial septal defects with a single Amplatzer device. AB - Transcatheter closure of single secundum atrial defects has become the standard of treatment. The purpose of our study was to analyze the results of using a single Amplatzer device for closure of double atrial septal defects. Such defects were diagnosed in 41 out of 363 patients with atrial septal defects (ASDs) closed by transcatheter method. In 39, a single Amplatzer device was used. The size of the larger defect ranged from 5 to 18 mm, the smaller defect from 2 to 7 mm, with the distance between the borders of the communications ranging from 2 to 12 mm. We performed sizing and closure of only the larger defect. The mean size of implanted devices was 16.5 +/- 5.5 mm, equal to the stretched diameter of the main defect or 1-4 mm larger. The closure rate assessed by color Doppler flow examination was 61% after 24 hr, 78% after 1 month, 83% after 3 months, 86% after 1 year, and 95% after 2 years. If the distance between the two defects exceeded 7 mm, residual leaks were observed but tended to decrease and disappear with time. Selected patients with double atrial septal defects can be effectively treated with implantation of a single Amplatzer device. PMID- 14755821 TI - Stent implantation in the ductus arteriosus for pulmonary blood supply in congenital heart disease. AB - Indications for catheter-based interventions in patent ductus arteriosus (PDA) dependent pulmonary blood flow have yet to be defined. The aim of this study was to assess the acute and midterm outcome after stent implantation in the PDA. Between 1996 and 2002, ductal stenting was performed in 21 neonates and infants (14 females/7 males) to maintain pulmonary blood supply in cyanotic congenital heart disease (CHD). Balloon-expandable stents were implanted in the PDA with a final diameter of 4-5 mm without procedural deaths. Stent patency was achieved for 8-1,130 days (median, 142). Reintervention was necessary in nine patients. Overall survival rate after 6 years was 86%. Two neonates died a few days after the procedure due to right heart failure not related to PDA stenting. Corrective surgery was possible in six patients. An additional aortopulmonary shunt was needed in three patients; Fontan type operations were performed in six. One patient died after bidirectional Glenn shunt, another five reached palliation by additional perforation of the atretic valve/balloon valvuloplasty, and two are awaiting surgery. We conclude that in many patients with cyanotic CHD, especially in those with ductal pulmonary perfusion and additional forward flow from the right ventricle, ductal stenting is an effective transcatheter approach. Morphology of the PDA predicts the risk of restenosis and necessity of reintervention. Growth of the pulmonary vascular bed allows corrective or palliative surgery, and some patients can be cured by the intervention alone. PMID- 14755822 TI - Mechanical thrombectomy using the AngioJect in a child with congenital heart disease. AB - Acute thrombosis of a vascular channel can be a serious and perhaps fatal complication. We describe a patient who developed complete thrombosis of the left pulmonary artery following catheter placement of a stent in this vessel shortly after open heart surgery. The thrombosis was successfully removed using the Possis AngioJet mechanical thrombectomy catheter. PMID- 14755823 TI - Native right ventricular outflow stent implantation in patients with unsalvageable extracardiac conduits. AB - We describe two patients with unsalvageable ventricular-to-pulmonary artery conduits in whom endovascular stents were implanted in the native right ventricular (RV) outflow providing double outlet ventricle and RV hypertension relief. Both patients are free from surgical reintervention at 7 years and 5 months, respectively. PMID- 14755824 TI - Transcatheter occlusion of baffle leaks following atrial switch procedures for transposition of the great vessels (d-TGV). AB - Baffle-related complications following atrial switch procedures for transposition are relatively common. Transcatheter treatment of baffle stenosis has an established role as a therapeutic modality. However, transcatheter device closure of atrial baffles leaks has rarely been reported. We report four patients who underwent device closure of baffle leaks using the Amplatzer septal occluder following atrial switch procedures in order to demonstrate the safety and utility of this method of treatment and to establish its role as a suitable alternative to surgical closure. PMID- 14755825 TI - Retrograde wire-assisted percutaneous transcatheter closure of persistent ductus arteriosus with Amplatzer duct occluder in the elderly: A new application. AB - Percutaneous transcatheter closure of persistent ductus arteriosus (PDA) has been well established in the pediatric field. For moderate- to large-sized PDA, the newly developed Amplatzer duct occluder had offered a good solution, but it depends on stiff wire and delivery sheath. We reported two elderly patients of PDA with vascular anatomy too difficult to be antegradely approached and were closed by a retrograde technique by an assisting wire from the descending aorta. The wire served as a guide and tracked the delivery system to cross the ductus from the venous side smoothly. This retrograde wire-assisted technique could be utilized to overcome the PDA of difficult vascular anatomy, which could not be easily fulfilled by conventional antegrade venous approach. PMID- 14755826 TI - Reactivity of the ductus arteriosus: Implications for transcatheter therapy. AB - We report our experience in a 13-month-old boy undergoing transcatheter coil occlusion of a patent ductus arteriosus. Constriction of the ductus arteriosus with subsequent relaxation resulted in inadvertent coil embolization. This case report and review of the literature have implications for transcatheter treatment of persistent ductus arteriosus. PMID- 14755827 TI - Stent implantation in right-sided patent ductus arteriosus to relieve severe cyanosis in adult patient with pulmonary atresia and ventricular septal defect. AB - Patients with unrepaired pulmonary atresia and ventricular septal defect may develop stenosis of collaterals or shunts to the pulmonary arteries leading to hypoperfusion of lungs and systemic hypoxemia. A 25-year-old female with pulmonary atresia and ventricular septal defect presented with progressively increasing cyanosis and exercise intolerance. A restrictive right-sided patent ductus arteriosus was identified as the main source of pulmonary blood flow. We report transcatheter implantation of a balloon-expandable stent across the stenosed duct to augment the pulmonary blood flow as a palliative management option. Patient had immediate improvement in arterial oxygen saturation from 66% to 85% with excellent clinical improvement and stable oxygen saturation on 8 months of follow-up. PMID- 14755828 TI - A "mouse in a trap" method for cerebral protection during carotid stenting: Technical note. AB - Carotid artery angioplasty and stenting (CAS) is now used as an alternative to surgical endarterectomy. The introduction of cerebral protection systems during stenting has improved carotid artery stenosis treatment, with less periprocedural complications. A "mouse in a trap" method was conceived and used in three patients. This involved an emboli entrapment-aspiration system using one proximal occluder in the common carotid artery and two distal occluders in the internal carotid artery or external carotid artery, followed by serial inflation-deflation cycles during each carotid stenting procedure. Debris was retrieved before dilation in one patient, after deployment in one, and after dilation in two. Although only used in a few cases to date, the method may improve the practice of CAS in treating patients with carotid stenosis, resulting in less thromboembolic events. PMID- 14755829 TI - Endovascular stent implantation for severe pulmonary artery stenosis in aortoarteritis (Takayasu's arteritis). AB - Symptomatic pulmonary artery stenosis is a relatively uncommon manifestation of aortoarteritis. We describe a patient of aortoarteritis with severe proximal right pulmonary artery stenosis who presented with dyspnea on exertion and central cyanosis. The pulmonary artery stenosis was successfully relieved by percutaneous transluminal balloon angioplasty and implantation of a balloon expandable stent. This resulted in immediate improvement in oxygen saturation, disappearance of cyanosis, relief of dyspnea, and marked improvement in right lung perfusion as demonstrated by pre- and postangioplasty technetium lung ventilation-perfusion scans. The changes in the pulmonary arterial wall morphology were detected precisely by intravascular ultrasound imaging. PMID- 14755830 TI - Stenting of anomalous left carotid artery using a radial artery approach. AB - A 90-year-old male presented with symptomatic severe stenosis of an anomalous left carotid artery originating from the brachicephalic trunk. A previous attempt at selective cannulation of the left carotid artery was unsuccessful using a transfemoral approach. We performed successful carotid artery angioplasty and stenting using a right radial artery approach. PMID- 14755831 TI - Absent dorsalis pedis pulse. PMID- 14755833 TI - Imaging a cognitive model of apraxia: the neural substrate of gesture-specific cognitive processes. AB - The present study aimed to ascertain the neuroanatomical basis of an influential neuropsychological model for upper limb apraxia [Rothi LJ, et al. The Neuropsychology of Action. 1997. Hove, UK: Psychology Press]. Regional cerebral blood flow was measured in healthy volunteers using H2 15O PET during performance of four tasks commonly used for testing upper limb apraxia, i.e., pantomime of familiar gestures on verbal command, imitation of familiar gestures, imitation of novel gestures, and an action-semantic task that consisted in matching objects for functional use. We also re-analysed data from a previous PET study in which we investigated the neural basis of the visual analysis of gestures. First, we found that two sets of discrete brain areas are predominantly engaged in the imitation of familiar and novel gestures, respectively. Segregated brain activation for novel gesture imitation concur with neuropsychological reports to support the hypothesis that knowledge about the organization of the human body mediates the transition from visual perception to motor execution when imitating novel gestures [Goldenberg Neuropsychologia 1995;33:63-72]. Second, conjunction analyses revealed distinctive neural bases for most of the gesture-specific cognitive processes proposed in this cognitive model of upper limb apraxia. However, a functional analysis of brain imaging data suggested that one single memory store may be used for "to-be-perceived" and "to-be-produced" gestural representations, departing from Rothi et al.'s proposal. Based on the above considerations, we suggest and discuss a revised model for upper limb apraxia that might best account for both brain imaging findings and neuropsychological dissociations reported in the apraxia literature. PMID- 14755834 TI - Rollvection versus linearvection: comparison of brain activations in PET. AB - We conducted a PET study to directly compare the differential effects of visual motion stimulation that induced either rollvection about the line of sight or forward linearvection along this axis in the same subjects. The main question was, whether the areas that respond to vection are identical or separate and distinct for rollvection and linearvection. Eleven healthy volunteers were exposed to large-field (100 degrees x 60 degrees ) visual motion stimulation consisting of (1) dots accelerating from a focus of expansion to the edge of the screen (forward linearvection) and (2) dots rotating counterclockwise in the frontal plane (clockwise rollvection). These two stimuli, which induced apparent self-motion in all subjects, were compared to each other and to a stationary visual pattern. Linearvection and rollvection led to bilateral activations of visual areas including medial parieto-occipital (PO), occipito-temporal (MT/V5), and ventral occipital (fusiform gyri) cortical areas, as well as superior parietal sites. Activations in the polar visual cortex around the calcarine sulcus (BA 17, BA 18) were larger and more significant during linearvection. Temporo-parietal sites displayed higher activity levels during rollvection. Differential activation of PO or MT/V5 was not found. Both stimuli led to simultaneous deactivations of retroinsular regions (more pronounced during linearvection); this is compatible with an inhibitory interaction between the visual and the vestibular systems for motion perception. PMID- 14755835 TI - The effect of stimulus intensity on brain responses evoked by transcranial magnetic stimulation. AB - To better understand the neuronal effects of transcranial magnetic stimulation (TMS), we studied how the TMS-evoked brain responses depend on stimulation intensity. We measured electroencephalographic (EEG) responses to motor-cortex TMS, estimated the intensity dependence of the overall brain response, and compared it to a theoretical model for the intensity dependence of the TMS-evoked neuronal activity. Left and right motor cortices of seven volunteers were stimulated at intensities of 60, 80, 100, and 120% of the motor threshold (MT). A figure-of-eight coil (diameter of each loop 4 cm) was used for focal stimulation. EEG was recorded with 60 scalp electrodes. The intensity of 60% of MT was sufficient to produce a distinct global mean field amplitude (GMFA) waveform in all subjects. The GMFA, reflecting the overall brain response, was composed of four peaks, appearing at 15 +/- 5 msec (Peak I), 44 +/- 10 msec (II), 102 +/- 18 msec (III), and 185 +/- 13 msec (IV). The peak amplitudes depended nonlinearly on intensity. This nonlinearity was most pronounced for Peaks I and II, whose amplitudes appeared to sample the initial part of the sigmoid-shaped curve modeling the strength of TMS-evoked neuronal activity. Although the response amplitude increased with stimulus intensity, scalp distributions of the potential were relatively similar for the four intensities. The results imply that TMS is able to evoke measurable brain activity at low stimulus intensities, probably significantly below 60% of MT. The shape of the response-stimulus intensity curve may be an indicator of the activation state of the brain. PMID- 14755836 TI - Neural mechanisms underlying reaching for remembered targets cued kinesthetically or visually in left or right hemispace. AB - Reaching for a target involves integrative coordinate transformation processes between the representation of the target location, the sensorimotor information of limb of reach, and body space. Although right hemisphere dominance for visuospatial information processing is well established, corresponding right hemisphere dominance for kinesthetic spatial information processing remains to be demonstrated. We explored neural mechanisms of encoding target locations using 15O-butanol positron emission tomography (PET) in normal volunteers in a factorial experiment, where modality (visual/kinesthetic) and hemispace of target presentation (left/right of midsagittal plane) were varied systematically. After target presentation, subjects reached to the encoded target location. PET data analysis using SPM99 showed increased neural activity (P < 0.05, corrected) associated with left hemispace target presentation in right hemisphere areas (sensorimotor, anterior cingulate, insular, and temporo-occipital cortex) only. By contrast, right hemispace target presentation activated bilateral temporo occipital cortex, which extended into the right temporo-parietal cortex and left sensorimotor cortex. A significant interaction of hemispace and modality of target presentation observed in right temporo-parietal cortex resulted from an increase in neural activity with kinesthetic target presentation in right hemispace. The data support an important role for the right temporo-parietal area in visuospatial processing and suggest a specific role of the right hemisphere in kinesthetic spatial processing. PMID- 14755837 TI - Different areas of human non-primary auditory cortex are activated by sounds with spatial and nonspatial properties. AB - In humans, neuroimaging studies have identified the planum temporale to be particularly responsive to both spatial and nonspatial attributes of sound. However, a functional segregation of the planum temporale along these acoustic dimensions has not been firmly established. We evaluated this scheme in a factorial design using modulated sounds that generated a percept of motion (spatial) or frequency modulation (nonspatial). In addition, these sounds were presented in the context of a motion detection and a frequency-modulation detection task to investigate the cortical effects of directing attention to different perceptual attributes of the sound. Motion produced stronger activation in the medial part of the planum temporale and frequency-modulation produced stronger activation in the lateral part of the planum temporale, as well as an additional non-primary area lateral to Heschl's gyrus. These separate subregions are consistent with the notion of divergent processing streams for spatial and nonspatial auditory information. Activation in the superior parietal cortex, putatively involved in the spatial pathway, was dependent on the task of motion detection and not simply on the presence of acoustic cues for motion. This finding suggests that the listening task is an important determinant of how the processing stream is engaged. PMID- 14755838 TI - Neurophysiological distinction of action words in the fronto-central cortex. AB - It has been suggested that the processing of action words referring to leg, arm, and face movements (e.g., to kick, to pick, to lick) leads to distinct patterns of neurophysiological activity. We addressed this issue using multi-channel EEG and beam-former estimates of distributed current sources within the head. The categories of leg-, arm-, and face-related words were carefully matched for important psycholinguistic factors, including word frequency, imageability, valence, and arousal, and evaluated in a behavioral study for their semantic associations. EEG was recorded from 64 scalp electrodes while stimuli were presented visually in a reading task. We applied a linear beam-former technique to obtain optimal estimates of the sources underlying the word-evoked potentials. These suggested differential activation in frontal areas of the cortex, including primary motor, pre-motor, and pre-frontal sites. Leg words activated dorsal fronto-parietal areas more strongly than face- or arm-related words, whereas face words produced more activity at left inferior-frontal sites. In the right hemisphere, arm-words activated lateral-frontal areas. We interpret the findings in the framework of a neurobiological model of language and discuss the possible role of mirror neurons in the premotor cortex in language processing. PMID- 14755840 TI - Back to nature. PMID- 14755839 TI - Reciprocal modulation and attenuation in the prefrontal cortex: an fMRI study on emotional-cognitive interaction. AB - Everyday and clinical experience demonstrate strong interactions between emotions and cognitions. Nevertheless the neural correlates underlying emotional-cognitive interaction remain unclear. Using event-related fMRI, we investigated BOLD-signal increases and decreases in medial and lateral prefrontal cortical regions during emotional and non-emotional judgment of photographs taken from the International Affective Picture System (IAPS). Emotional and non-emotional judgment conditions were compared to each other as well as with baseline allowing for distinction between relative signal changes (comparison between conditions) and true signal changes (referring to baseline). We have found that: (1) both emotional and non emotional judgment of IAPS pictures were characterized by signal increases in ventrally and dorsally located lateral prefrontal cortical areas and concurrent signal decreases in ventro- and dorsomedial prefrontal cortex; (2) direct comparison between emotional and non-emotional judgment showed relative signal increases in ventro- and dorsomedial prefrontal cortex, and in contrast, relative signal increases were detected in ventrally and dorsally located lateral prefrontal cortical areas when comparing non-emotional to emotional judgment; and (3) as shown in separate comparisons with baseline, these relative signal changes were due to smaller signal decreases in ventro- and dorsomedial prefrontal cortex and smaller signal increases in ventrally and dorsally located lateral prefrontal cortical areas during emotional judgment. Therefore, the emotional load of a cognitive task lead to both less deactivation of medial prefrontal regions and, at the same time, less activation of lateral prefrontal regions. Analogous patterns of reciprocal modulation and attenuation have previously been described for other cortical regions such as visual and auditory areas. Reciprocal modulation and attenuation in medial and lateral prefrontal cortex might constitute the neurophysiologic basis for emotional-cognitive interaction as observed in both healthy and psychiatric subjects. PMID- 14755841 TI - Welcome to the ward? PMID- 14755842 TI - Art for art's sake. PMID- 14755843 TI - Talking cure. PMID- 14755845 TI - A window of opportunity. PMID- 14755846 TI - Loose nuts. PMID- 14755847 TI - Research. PMID- 14755849 TI - Physician's misinterpretation of HIV test report leads to liability. Doe v. Arts. PMID- 14755848 TI - The C3435T MDR1 gene polymorphism is not associated with susceptibility for ulcerative colitis in Greek population. PMID- 14755850 TI - Florida court refuses to enforce noncompete clause. PMID- 14755851 TI - [Selenium in selected species of mushrooms from Poland]. AB - The selenium was quantified in the caps, stalks or a whole fruiting bodies of king bolete (Boletus edulis), brown birch scaber stalk (Leccinum scabrum), parasol mushroom (Macrolepiota procera), fly agaric (Amanita muscaria) and poison pax (Paxillus involutus) collected at the various regions of Poland in 1998-2001. King bolete, parasol mushroom and fly agaric were a much more abundant in selenium than brown birch scaber stalk or poison pax. Some differences were observed between the selenium content of the particular species collected at different sites as well as depending on anatomical part of the fruiting body. PMID- 14755852 TI - [Radon-222 in drinking water of Karkonoskie Plateau]. AB - Radon-222 concentration in surface water, wells water and tap water in the main towns and villages which are located in area of Karkonoskie Plateau has been quantitative determined. The measurements were performed using the alpha liquid scintillation counting method. Majority of waterworks in Karkonoskie Plateau is supplied with the ground water in which the radon concentration is high from 87.5 Bq/l to 818.1 Bq/l. The waterworks in Karpacz are supplied with the surface water, which main characteristic is low radon concentration (below 10 Bq/l) and with the ground water have a high radon concentration (to 541 Bq/l). Radon-222 concentration in water of individual wells was similar to concentration in the ground water. PMID- 14755853 TI - [Fatty acids content in selected edible oils]. AB - The four edible oils purchased on Warsaw marked in the years 1996-2002 were investigated. We confirmed that investigated oils contained relatively small amount of saturated fatty acids (7.0-13.4%) and only traces of trans isomers. In the same time they were characterized with high content unsaturated fatty acids, both mono- and polyunsaturated. The highest content of monounsaturated acids (65%) characterized rapeseed oil, whereas polyunsaturated ones--sunflower oil (68%). Polyunsaturated acids were represented mainly by linoleic acid (C18:2). The results of investigation show that oils available on the market despite of their different manufacturers ha got the good health value. PMID- 14755854 TI - [Microbiological quality of muesli samples purchased in retail network]. AB - The estimation of microbiological quality of muesli samples was the aim of this investigation. The study included 40 samples which represented various consignments of muesli, produced in 2002 year. Total number of aerobic mesophilic bacteria, yeasts and moulds, and also occurrence of pathogenic bacteria, coliforms and enterococci were determined. The systematic units of muesli fungal flora have been identified. It was stated that microbiological quality of all tested muesli was good. No pathogenic bacteria (Salmonella, Staphylococcus aureus and Bacillus cereus) was detected and also coliforms and enterococci titre was correct (> 0.1 g). A majority of samples characterized low contamination levels of mesophilic aerobic bacteria and fungi, and their total number did not exceed, suitably 10(3) and 10(2) cfu/g. The mesophilic aerobic bacteria were mainly represented by vegetative forms. The average share of these bacteria spores in total number of bacteria received 30%. The Aspergillus sp. and Penicillium sp. were regular elements of muesli fungal flora. PMID- 14755855 TI - [Effect of technological process in bottled water production on its redox properties]. AB - The redox potential was measured in selected natural mineral and spring waters of different origin and chemical composition. To analyse the effect of the technological process the samples to be studied were collected at different stages of the bottling process: directly at the intake, at the beginning of the pipeline, after the deironing and/or demanganisation, after filtration, at the end of the pipeline and at the end of the process from the freshly filled bottles. Along with the redox potential measurements, some physico-chemical parameters of the samples studied were determined: colour, turbidity, conductivity, pH value, susceptibility to oxidation, contents of iron, manganese, antimony and arsenic. The potentiometric method applied to measure the redox potential was found suitable for the purpose of the study. PMID- 14755856 TI - [Comparative assessment of vitamin C content and evaluation of energy intake in children daily food rations]. AB - The aim of this study was comparative assessment of content of vitamin C in daily food rations orphans children aged 1-6 years by using two methods--analytical and calculation and evaluation of energy intake by calculation. The levels of energy and vitamin C intake in daily food rations were calculated using menu data and data of food composition tables and compared with the recommended dietary intake (RDI). In addition vitamin C was determined in daily food rations by high performance liquid chromatography method (HPLC). Differences between two methods for evaluation of content of vitamin C were not statistically significant. Contribution of energy to individual meals and share of main nutritive components in supply of energy to daily food rations did not correspond with recommendations. Beyond this daily food rations orphans children were plentiful, monotonous and uniform without using seasonal products. PMID- 14755857 TI - [Influence of carcinogenesis in the oral cavity on the level of some bioelements in the saliva]. AB - The aim of this study was the examination of influence the oncogenesis process of oral cavity on the level of some bioelements (Ca, Mg, K, Na, Zn, Cu, Mn, Fe) in the saliva. The saliva was sourced from patients with oral cancer (carcinoma planoepitheliale spinocellulare keratodes and akeratodes: mucosae buccae, fundi oris, linguae, palati molli). The determination of the analyzed elements were made by atomic absorption spectrophotometry method. The levels of most of the analyzed elements (Ca, Mg, Na, Zn, Cu, Mn and Fe) were significantly higher in the case grop comparing to controls. PMID- 14755858 TI - [Relative validity of self-assessment of silhouette and BMI (body mass index)]. AB - Overweight and obesity in childhood and adolescence is unquestionable risk factor for pathogenic obesity in adulthood, high mortality and morbidity for cardiovascular diseases and other health disorders, and also may cause the worse social and economical adaptation. Nevertheless, little is yet known about subjective perception of own body, the pathway leading to dissatisfaction of the body, development of chronic stress and behavioural disorders (anorexia, binge eating, bulimia) as a consequence. In Health Promotion Department of the National Institute of Hygiene the multidimensional investigations of adolescents' health and life style were undertaken, and analysis of association between subjective image of body and real body mass was a part of these investigations. Data were obtained from 672 randomly selected schoolchildren aged 14-15 years attending seventeen public and private schools in Warsaw. Respondents informed about their weight and high for calculation BMI. Simultaneously, they were asked, whether they assess themselves as leaner than their peers, the same or thicker. The study showed that girls in comparison with boys more accurately assessed their silhouette. The boys were more likely than girls to perceive themselves as the same as their peers, despite they had real underweight or overweight. Our findings suggest that real mass of body itself account for variance of subjective perceived body in moderate degree, and there are other factors influencing body image at least as real mass of body. PMID- 14755859 TI - [Acoustic characteristics of classrooms]. AB - Quality and usefulness of school rooms for transmission of verbal information depends on the two basic parameters: form and quantity of the reverberation time, and profitable line measurements of school rooms from the acoustic point of view. An analysis of the above-mentioned parameters in 48 class rooms and two gymnasiums in schools, which were built in different periods, shows that the most important problem is connected with too long reverberation time and inappropriate acoustic proportions. In schools built in the 1970s, the length of reverberation time is mostly within a low frequency band, while in schools built contemporarily, the maximum length of disappearance time takes place in a quite wide band of 250-2000 Hz. This exceeds optimal values for that kind of rooms at least twice, and five times in the newly built school. A long reverberation time is connected with a low acoustic absorption of school rooms. Moreover, school rooms are characterised by inappropriate acoustic proportions. The classrooms, in their relation to the height, are too long and too wide. It is connected with deterioration of the transmission of verbal information. The data show that this transmission is unequal. Automatically, it leads to a speech disturbance and difficulties with understanding. There is the need for adaptation of school rooms through increase of an acoustic absorption. PMID- 14755860 TI - [Air microflora of university cafeteria]. AB - The numbers of aerobic mesophilic bacteria, yeasts and moulds were obtained by sedimentation method. The investigation included six areas, which have been separated on the ground of their function: 1. washing and peeling of potatoes and vegetables, 2. initial treatment of raw materials, 3. washing up of kitchen utensils, 4. cooking of meals, 5. serving of meals, 6. dining room. The samples of air were collected in 32 investigation points in the morning (8-8(30)) and in the afternoon (15-15(30)). Twelve series of measurements were carried out and in general 768 of air samples were tested. The results show that numbers of bacteria, moulds and yeasts were variable and received respectively 75-4550, 0 4565 and 0-290 cfu/m3. Analysis of variance proved that differences between mean number of microorganisms in the air were significant in dependence on the kind of place, time of a day and series of measurements. In the morning the highest microbiological contamination characterized the air of "washing and peeling" area. In the afternoon the number of all groups of microorganisms in the air of 1 and 2 areas was reduced. In the other places the amount of bacteria and yeasts increased, but mean number of moulds was reduced. Respectively 3% and almost 20% of air tested samples not answered for bacteria and fungi numbers recommended to kitchen areas. Filamentous fungi were represented mainly by Cladosporium sp. and Penicillium sp. PMID- 14755862 TI - Managed medications and the new Medicare. PMID- 14755863 TI - Bluish discoloration of periorbital area. Eyelid purpura and patient's medical history lead to diagnosis. PMID- 14755861 TI - [Release of volatile organic compounds from textile floor coverings in higher temperatures]. AB - The effect of temperatures 23 degrees C, 29 degrees C, and 50 degrees C on emission of 4-phenylcyclohexene (4-PC), styrene, total volatile organic compounds (VOCs) and formaldehyde from textile floor coverings with textile backing and styrene/butadiene latex precoat was examined. Tested coverings didn't show emission of styrene and formaldehyde. At 23 degrees C and 29 degrees C two tested coverings showed emission of 4-PC on low level. The higher emission was observed from all coverings at 50 degrees C. VOCs emission from coverings increased with increasing of temperature, especially at 50 degrees C. In conclusion some of new textile floor coverings can cause contamination of indoor air after application of sub-floor heating. Contamination, however, will decrease with time. PMID- 14755864 TI - Sleep, health, and aging.... PMID- 14755865 TI - Sleep, health, and aging. Bridging the gap between science and clinical practice. AB - Problems with sleep are common with advancing years and occur in over half of adults age 65 and older. It has been estimated that insomnia affects about a third of the older population in this country. This inability to have restful sleep at night results in excessive daytime sleepiness, attention and memory problems, depressed mood, falls, and lowered quality of life. Other factors associated with aging, such as disease, changes in environment, or concurrent age related processes also may contribute to problems of sleep. Data indicate that age by itself does not predict incident complaints of insomnia, even in the presence of lowered sleep efficiency and decreased proportion of slow-wave sleep. Rather, the prevalence of insomnia and other sleep disorders is high in the geriatric population due to the associated comorbidities common in late life. It is now evident that disturbance in sleep can also lead to adverse changes in functioning of a number of body systems. PMID- 14755866 TI - Patient handout. Sleep talk with your doctor. PMID- 14755867 TI - Sleep disorders in older adults. A primary care guide to assessing 4 common sleep problems in geriatric patients. AB - Complaints about sleep are common among older adults, yet are not a normal consequence of old age. Disordered sleep can be due to medical conditions, chronic diseases, psychiatric disorders, and medications. Age-associated, oropharyngeal anatomical changes can result in sleep-disordered breathing (sleep apnea). In addition, the circadian rhythm advances that accompany aging can cause early evening lethargy and early morning awakenings. Discovering and treating the underlying cause(s) of the sleep disorder is the first step in approaching the problem. PMID- 14755868 TI - By way of an introduction. Paving the way for a psychiatric referral. PMID- 14755869 TI - Aggressive lipid-lowering regimen halts CHD progression. PMID- 14755870 TI - Valsartan an alternative to ACE inhibitor in high-risk MI. PMID- 14755871 TI - Acute sinusitis. When--and when not--to prescribe antibiotics. AB - Clinical diagnosis of acute sinusitis is troublesome because it involves use of a cluster of diagnostic criteria that have only moderate sensitivity. Ancillary testing with radiography or antral puncture is impractical, expensive, and usually unnecessary in the primary care setting. Antibiotic therapy is not beneficial for most patients in whom acute sinusitis is suspected, even when radiographic abnormalities are found. Simple management algorithms and patient information are now available to aid primary care physicians in offering appropriate therapeutic measures and reassuring patients who are expecting "'a pill for every ill' when that pill is an antibacterial." PMID- 14755872 TI - Getting tough with metabolic syndrome. PMID- 14755873 TI - Primary prevention of cirrhosis. Public health strategies that can make a difference. AB - Cirrhosis is associated with high morbidity and mortality and often affects persons during the most productive years of life. In the United States, alcoholic liver disease is the leading contributor to the overall prevalence of cirrhosis, followed by infection with hepatitis B virus (HBV) and hepatitis C virus (HCV). In this article, Drs Karsan, Rojter, and Saab examine lifestyle behaviors that can lead to cirrhosis and enumerate public health strategies aimed at primary prevention. PMID- 14755874 TI - Can vaccination prevent sinusitis? PMID- 14755875 TI - Improved therapy for atopic eczema. New immunomodulators clear the rash with few side effects. PMID- 14755876 TI - What role for weight-loss medication? Weighing the pros and cons for obese patients. AB - Obesity is a chronic medical disorder that is not going away anytime soon. Physicians need all the education, tools, and resources possible to successfully help their overweight and obese patients. Weight-loss medications alone are clearly not the answer. However, they are one tool physicians can use in combination with lifestyle changes to increase the success of long-term weight loss in selected patients. PMID- 14755877 TI - Bariatric surgery. For the right patient, procedure can be effective. AB - Obesity is a chronic health problem with associated medical complications involving multiple physiologic systems. Some obese patients succeed in losing weight with traditional, nonsurgical therapies and interventions. In this article, Drs Mattison and Jensen discuss the option of bariatric surgery for those patients who have not responded to medically sound weight-loss programs and for whom the complications of obesity pose a high and ongoing risk. PMID- 14755878 TI - Tackling obesity in a 15-minute office visit. Physicians can start patients on an effective weight-loss program, despite time constraints. AB - Primary care physicians can provide their patients with a variety of practical strategies to combat obesity regardless of the time constraints experienced in today's busy clinical practice setting. Keys to success include creating an individualized approach based on each patient's specific health risks and habits, using various resources (e.g., support groups, registered dietitians), and encouraging and empowering patients to become active participants in their weight loss program. PMID- 14755880 TI - Patient notes: organ donation. PMID- 14755879 TI - Is it andropause? Recognizing androgen deficiency in aging men. AB - In primary care practice, it is not unusual to encounter male patients in their 50s or older who report having loss of libido, erectile dysfunction, fatigue, and depression. Such signs and symptoms may signal an age-related decline in androgen levels, which commonly begins after age 40. However, psychologic problems and medical illness often confound the diagnosis. Drs Tan and Pu, who are currently conducting research on androgen deficiency, discuss the diagnostic difficulties of the physiologic phenomenon of andropause and offer a comprehensive approach to clinical assessment and laboratory evaluation. PMID- 14755882 TI - Molecular therapeutics: are we making progress? PMID- 14755883 TI - DNA microarrays in vaccine research. AB - DNA microarrays represent the technology platform for a wide variety of analytical methods built around the detection of sequence-specific nucleic acid hybridization. They allow the analysis of complex biological systems on the basis of the genome and transcriptome with high simplicity, efficiency, sensitivity and specificity. This positions DNA microarrays as ideal tools for novel approaches in biomedical research and explains the tremendous pace at which they are being applied to an enormous variety of systems and projects. Recently, vaccine researchers began to use DNA microarrays for the identification of vaccine candidates against bacterial, parasitic and viral pathogens, as well as tumors. Here, DNA microarrays as a technology platform in these areas of vaccine research are reviewed and their limitations, as well as their potential for future applications in vaccinology, critically evaluated. PMID- 14755884 TI - Serum protein fingerprinting. AB - The core technologies in the rapidly expanding field of proteomics have matured to the point where quantitative measurements of thousands of proteins can be conducted, enabling truly global measurements of protein expression. This advent has brought with it the hope of discovering novel biomarkers that promise a renaissance in clinical medicine. To meet this need, many proteomic studies have focused on the identification and subsequent comparative analysis of the thousands of proteins that populate complex biological systems such as serum and tissues. A novel application of mass spectrometry has been in proteomic pattern analysis, which has emerged as an effective method for the early diagnosis of diseases. In stark contrast to 'classical' proteomics, proteomic pattern analysis relies on the pattern of proteins observed, rather than on the discrete identification of a protein. Proteomic pattern technology allows hundreds of clinical samples to be analyzed per day and promises to be a novel, highly sensitive predictive clinical tool to improve diagnostic and prognostic medicine. PMID- 14755885 TI - In vivo imaging in the development of gene therapy vectors. AB - The availability of imaging techniques and reporter probes that provide information regarding the biodistribution of gene therapy vectors and expression of encoded genes in living animals, has the potential to accelerate the development and testing of new gene therapies. Techniques using novel imaging markers and labeled reporter molecules enable longitudinal studies that provide quantitative or semi-quantitative data reflecting gene expression regulation in living animals, which may significantly assist in the development of therapeutics targeted to individual organs or cell populations. Non-invasive imaging studies performed in animal models of human disease are crucial for translational research for future clinical development. PMID- 14755886 TI - Integrins and angiogenesis: unlocking the route to gene therapy. AB - Angiogenesis is defined as the process of vascularization of a tissue, involving the development of new capillary blood vessels. Both the induction and inhibition of this process can have therapeutic benefits in various pathological conditions. Integrins are a structurally elaborate family of adhesion molecules; they participate in a wide range of biological processes, including angiogenesis. Endothelial cells are intimately involved in angiogenesis and are thought to mediate this function partially through the integrins on their cell membrane, which regulate cell-cell and cell-matrix contacts. Extensive research into elucidating the mechanisms involved in the angiogenesis process have led to the discovery of a growing number of genes encoding pro- and anti-angiogenic proteins. A variety of gene therapy approaches have been used to deliver many of these genes to induce or inhibit the angiogenesis process with varying levels of success. This review investigates whether targeting gene therapy vectors to integrin receptors found on endothelial cells is a viable means to improve the efficiency of the gene transfer process. PMID- 14755887 TI - Adenoviral p53 gene therapy in squamous cell cancer of the head and neck region. AB - Treatment options for recurrent head and neck squamous cell carcinoma are limited. Morbidity and mortality are largely related to local recurrence. Experimental investigation of adenoviral p53 gene therapy is being explored as a new therapeutic modality for patients with recurrent squamous cell carcinoma in the head and neck region. The function of the p53 gene is to maintain the genetic integrity of the cell and induce apoptosis when DNA damage is irreparable. Phase I trial results indicate that multiple injections of up to 2.5 x 10(12) viral particles per injection administered during a 3-week cycle are well tolerated and demonstrate evidence of local regional control. Dose-related activity correlating with transient survival advantage is demonstrated in a meta-analysis of phase II trials. This review will summarize phase I and II trial results. Conclusions drawn from these studies justify the currently ongoing phase III investigation. PMID- 14755888 TI - Overview of phase I studies of intravenous administration of PV701, an oncolytic virus. AB - PV701 is an attenuated, non-recombinant, oncolytic strain of Newcastle disease virus that displays preclinical intravenous (i.v.) efficacy. PV701 selectively lyses tumor cells versus normal cells based on tumor-specific defects in the interferon-mediated antiviral response. In three phase I trials in 113 patients, the effects of dose, schedule and i.v. infusion rate were evaluated. Three types of adverse events were seen: flu-like, tumor-site-specific and those occurring during infusion. The first PV701 dose desensitized the patient to the side effects of further doses, allowing a 5- to 10-fold increase in the maximum tolerated dose for subsequent doses compared with the first dose. Tumor responses were first noted at the higher doses achieved using desensitization. In 95 evaluable patients, there were ten responses (six major and four minor), with five of these responses occurring in the most recent trial of 18 patients that employed desensitization, high repeat doses and a slower infusion rate. Phase II studies are planned. PMID- 14755890 TI - Defining the human targets of phorbol ester and diacylglycerol. AB - Phorbol esters (PEs) and their derivatives are potent tumor-promoting agents. The best known receptors for these substances are the novel and classical isotypes of protein kinase C (PKC), which bind PE and the physiological second messenger diacylglycerol (DAG) by cysteine-rich domains, the C1 domains. However, PKC is not the sole receptor of PE, a concept that has been largely ignored in the past. PE (in addition to DAG) also targets C1-containing receptors unrelated to PKC. In order to get a better insight into DAG/PE-mediated signaling and the pathways involved, it is necessary to first determine all ligand-interacting proteins. Employing various sources of data, 66 different C1-containing human proteins are presented and predictions of their DAG/PE-binding potential are attempted. Defining the entire set of key mediators for the physiological DAG responses and for PE-induced tumorigenesis may aid our understanding of signal integration and can also help to design new strategies for therapeutic cancer intervention. PMID- 14755889 TI - Discovery and validation of new molecular targets for ovarian cancer. AB - The occult progression of tumors within the peritoneal cavity results in the late diagnosis and high mortality rate of ovarian cancer. An improved understanding of the molecular biology of ovarian cancer has led to the discovery of novel molecules as targets for the treatment of ovarian cancer. This review highlights the latest advance of mucins, which are upregulated in cancer cells and interact with the epidermal growth factor receptor family to regulate cell behavior via signaling pathways associated with malignant transformation, invasion and metastasis. Disruption of these molecules might represent a novel therapeutic approach in treating ovarian cancer. We also describe the recent development and validation of the most studied mucins (MUC1 and MUC16), ErbB-2 (Her2/neu) and folate binding protein as target-based immunotherapy of ovarian cancer. PMID- 14755891 TI - Pharmacogenomics of psychiatric drug treatment. AB - Clinical outcome to psychiatric drug therapy varies widely among patients. A significant proportion fail to respond satisfactorily, while others react adversely to these compounds. Multiple factors account for these inter-individual differences, however, the most significant contribution is the influence conferred by genetically inherited traits. Pharmacogenetics has been at the forefront of work to delineate this influence, by identifying polymorphisms within candidate genes involved in drug metabolism, neurotransmission and signal transduction that are important to the actions of these drugs. Progress in biomolecular technology has paved the way for pharmacogenomic strategies that examine this influence on a genomic level, and look set to perform the task of candidate gene identification more expeditiously. Microarray expression profiling is one particularly important pharmacogenomic development that has immense potential, and its use has already facilitated the identification of synaptic genes and other diverse candidates that appear relevant to the actions of these drugs. Knowledge from these studies will ultimately lead to the individualization of psychiatric drug treatment, while providing new insights into the etiology of these disorders and their future treatment. PMID- 14755892 TI - Exploiting the hypoxia response. AB - Hypoxia (low oxygen) is a defining physiological feature of a number of diseases, including cancer, cardiovascular disease and retinopathy. Hypoxia plays an active role in the pathology of these diseases through its impact on gene expression, thereby making the hypoxia-signaling pathway a key target for the development of novel molecular therapies. This review focuses on how the elucidation of this pathway has led to the development of novel therapeutic strategies, including physiologically targeted gene therapy and the identification of novel therapeutic targets within the hypoxia-signaling pathway. PMID- 14755893 TI - Technology evaluation: bevacizumab, Genentech/Roche. AB - Bevacizumab, an antivascular endothelial growth factor monoclonal antibody, is being developed by Genentech and Roche as an anti-angiogenesis therapy for the potential treatment of solid tumors. In June 2003, bevacizumab was granted Fast Track status by the FDA for the potential treatment of first-line colorectal cancer. The antibody is currently in phase III trials for non-small-cell lung, colorectal and breast cancers, and in phase II trials for various other solid tumor types. PMID- 14755894 TI - Technology evaluation: Onyvax-105, Onyvax. AB - Onyvax, under license from the Cancer Research Campaign, is developing Onyvax-105 (105AD7), an anti-idiotype monoclonal antibody, for the potential treatment of colorectal cancer. Onyvax initiated phase II clinical trials in May 2000. PMID- 14755895 TI - Technology evaluation: pegaptanib, Eyetech/Pfizer. AB - Eyetech Pharmaceuticals and Pfizer are co-developing the anti-vascular endothelial growth factor aptamer, pegaptanib, as an angiogenesis inhibitor for the potential treatment of age-related macular degeneration and diabetic macular edema, in addition to other ocular diseases. Gilead was previously investigating the aptamer for the potential treatment of cancer, however, no data have been published for this indication since 1999. PMID- 14755896 TI - Technology evaluation: HSPPC-96, antigenics. AB - Antigenics is developing HSPPC-96, a vaccine based on heat shock protein for the potential treatment of various cancer indications. The compound is currently undergoing phase III clinical trials for renal cell carcinoma and melanoma. PMID- 14755897 TI - [Thromboembolic venous disease]. PMID- 14755898 TI - [Pulmonary thromboembolism: long term course and clinical epidemiology]. AB - BACKGROUND: Long-term clinical course of pulmonary thromboembolism is not well known. Our aim was to know the events which occur to in-patients diagnosed of pulmonary embolism. METHODS AND PATIENTS: This is a prospective observational study from May-92 to December-2002 with all in-patients diagnosed of pulmonary thromboembolism at a clinical area of Internal Medicine. Main targets were to know survival, relapses, major hemorrhage rate (Defined as those episodes of bleeding which needed blood transfusion and readmission) and cancer associated rate (Previous and newly diagnosed cancer). Follow up were carried out with telephone contacts with patients and relatives in case of death, and with the computerized system of patients and clinical events of Health Service of Navarra. RESULTS: One hundred and sixteen patients were included in the study (Mean age 72 SD 11 years male 54%). During index episode 4 (3.7%) patients dead. Ten patients were lost in follow up. The rest 102 patients were traced for 31.81 SD 31.23 months (Range 1-127). Relapse rate was 19.6% that occurred 22.64 SD 24.57 (Range 1-73) months after index episode (Twelve pulmonary embolisms, 5 deep venous thromboses and 3 sudden death with dyspnea). Major hemorrhage rate was 10.4%. During follow up 14 (13.7%) new cancers were diagnosed (Lung 4, prostate 2, bladder 2, and colorectal, ovary, breast, liver and kidney one each one). At all prevalence of cancer associated with pulmonary thromboembolism was 31%. Mortality rate was 37% (Men 25%, women 49%, p < 0.01). Main causes of death were cancer (32%) and relapse of pulmonary thromboembolism when joined with treatment complications 24%. Half of deaths occurred in the first year of follow up, showing a shortened survival those patients with cancer (p = 0.02) and patients with relapses of pulmonary embolism (p = 0.06). Beyond the first year, mortality declines to a rate of 10% per year mainly because of cardiovascular causes. Mortality associated factors were age > 75 years (p < 0.001) gender female (p < 0.01), a delayed admission and treatment from the beginning of symptoms (p < 0.05), higher LDH level (p < 0.01) and coexistence of cancer (p < 0.05). In logistic-regression analysis age, delayed admission and treatment and higher LDH levels were predictors of long-term death. CONCLUSIONS: Patients with pulmonary embolism show a high mortality rate, with a critical period during the first year after index episode, being deaths associated to cancer and to a composite of relapse of venous thromboembolic disease and bleeding complications. Mortality rate beyond the first year declines, being deaths explained because of cardiovascular causes. An advanced age, a delayed diagnosis and treatment and serum LDH may predict long-term mortality. PMID- 14755899 TI - [Seasonal changes in morbimortality caused by pulmonary thromboembolism in Galicia]. AB - OBJECTIVES: The goal of this report was to analyze the seasonal variation in the pulmonary embolism (EP) hospitalizations and mortality at the hospitals of the Galician Service of Health (SERGAS) in the Northwest of Spain, during the period 1996-2001. METHODS: The Related Group of Diagnosis (DRG) 78 (pulmonary embolism) from the SERGAS hospital discharge dataset during the 6-year period was examined. Age, sex, month of discharge and seasonal period was annotated. RESULTS: A total of 2831 hospitalizations for TEP were recorded (44% males, mean inhospital stay: 12.8 days, 70% > 65 years). Monthly median was 38 discharges and the variations above and below this were +26% (December) and -25%) (July), respectively. There were more frequents hospitalizations in autumn (OR:1.35, 95%CI:1.19-1.53) and winter (OR: 1.29, CI: 1.14-1.46). Mortality peaked in spring (OR:2.18, CI: 1.18 4.05). Nearly 83% of these deaths occurred in persons > 75 years old. CONCLUSIONS: We observed a marked increase in EP hospitalizations during colder months and in mortality during spring. These observations may have implications in the adoption of preventive measures. PMID- 14755900 TI - [Clinical analysis of 91 cases of median and small vessel vasculitis in adults]. AB - OBJECTIVE: To evaluate the clinical and biological differences between medium sized vessel vasculitis and small vessel vasculitis. PATIENTS AND METHODS: descriptive and retrospective study of 91 patients with vasculitis attended in our hospital from January 1991 to mars 2001. We describe the characteristics of clinical and analytic features. RESULTS: 57% were males. The mean age was 61.9 +/ 18.6 years (17 to 90 years). The symptoms and affected organs were: palpable purpura (89%), fever (36%), asthenia (20%), arthromyalgias (19%), nephropathy (18%), arthritis (16%), abdominal pain (16%), neuropathy (8.7%), pulmonary involvement (6.5%). 25% had several episodes, lasting clinical, chronic disease, 42% had evidence of two or more involve organs. The patients with pauci-inmune vasculitis presented more asthenia, nephritis, pulmonary involvement, multi organic involvement and mortality related to the process. We did not found significant differences respect to the others clinical manifestations analysed. CONCLUSIONS: There is a substantial overlap among different vasculitis, the presence or absence of some clinical and biological features can help in the differentiation and characterization of the different entitles. PMID- 14755901 TI - [Spinal epidural involvement as presentation form of non-Hodgkin's lymphoma: report of 6 cases]. AB - Spinal cord compression and radicular involvement are infrequent events in the natural history of non-Hodgkin's lymphoma, which usually are secondary to invasion of spinal extradural space, and rarely are the presenting manifestations of this condition. Thoracic segment is predominantly affected, but any spinal region can be affected. Histologically, the spinal tumour are frequently a high grade B lymphoma with aggressive behaviour. The aim of this work was to describe six new cases of non-Hodgkin's lymphoma in adults (five woman and one men), presenting with spinal neurological complications. The medical literature about this entity are revised. PMID- 14755902 TI - [Brunner gland hamartoma and anisakiasis: etiologic association?]. AB - Brunner's gland hamartoma is a rare duodenal tumor generally localized in the duodenal bulb. Normally assymptomatic, it might cause upper gastrointestinal bleeding or intestinal obstruction. The diagnosis is based on upper gastrointestinal endoscopic or barium examination findings, and its treatment includes surgical or endoscopic resection, with an optimum prognosis. We present the case of a 59-year-old woman who was admitted to the hospital with an upper gastrointestinal bleeding picture. Endoscopic examination showed an ulcerated polyp in duodenal bulb which was cut with polipectomy wire. Histological slides showed a parasitic granuloma within a Brunner's gland hamartoma. Skin prick test and specific IgE determination were positive for Anisakis. Up to our knowledge, this association has never been described before. PMID- 14755903 TI - [Extrahepatic portal hypertension: spleno-portal thrombosis secondary to protein C deficiency]. AB - Portal vein thrombosis (PVT) is the most frequent cause of hypertension portal extrahepatic. It is a rare disorder an the main risk factors are cirrhosis, hepatobiliary malignancies and prothrombotic disorders, which have been identified as major risk. Therapy with anticoagulants must to be considered in acute portal thrombosis or chronic one and proven hypercoagulability. We present the case of a twenty-nine years old patient, with extrahepatic portal hypertension secondary to portal and splenic vein thrombosis, who was diagnosed because of splenomegaly and a coagulation disorder. A protein C deficiency were discovered and anticoagulation and beta-blocker therapy were initiated. One year later the patient had not presented complications concerning to the disease or to the treatment. PMID- 14755904 TI - [Strongyloides stercoralis in the south of Galicia]. AB - Strongyloides stercoralis is the only parasite which can produce a chronic illness in humans, being through autoinfection. This nematode can also provoke death when patient's immunologic state deteriorates producing a massive hyper infection. The first patient with strongyloidiasis who has always lived in the Galician South area is described. The clinical picture consisted of unspecific cutaneous lesions and abdominal pain with severe peripheral eosinophilia (> 20,000/ml. The diagnosis was carried out observing the larvae in the fecal examination and was confirmed with a culture. Treatment with albendazole failed and the healing was reached with ivermectin. We must consider the possibility of strongyloidiasis because misdiagnosing these patients as eosinophilic gastroenteritis there would be a higher risk of hyperinfection if they are treated with corticosteroids. PMID- 14755905 TI - [Chin numbness as initial manifestation of systemic cancer]. AB - This issue describe the clinical picture of a man who consulted by hypoanaesthesia in the chin area that was secondary to a lung cancer. The most common primary cancers associated with mental neuropathy are haematological malignancies, breast and lung cancers. The presence o mental neuropathy must not be considered an "banal" symptom. It should be quickly recognised in order to research the presence of cancer. PMID- 14755906 TI - [New advances in the knowledge on post-thrombotic syndrome]. AB - The real incidence of the post-thrombotic syndrome (PTS) is not known precisely, though of the most part of the variable studies, seems be deduced that it can be established a year after the deep venous thrombosis (DVT) acute of the inferior members in 17% to the 50% of the patients. Inseparably united to the venous hypertension that continues to the development of the incompetence valvular, is accompanied of a series of inflammatory reactions that include the increase in the permeability endothelial, the union of the circulating leukocytes at endothelium, the infiltration by monocytes, lymphocytes and mastocytes of the connective tissue, and the development of infiltrated tissular fibrotics and different molecular markers. To the contrary that in the DVT, we know very little about the factors that increase the risk of suffering a PTS, since the only one identified up until now it is the recurrent DVT. Currently we have different scales standardized for their your clinical diagnosis, though the Echo-Doppler is, currently, the technique not invasive of election to detect, locate and evaluate the venous disability valvular and the venous obstruction chronicle. The modern technical of image: computed tomography (CT), magnetic resonance (MR) and isotopics have a promising future, even though are found in validation phase. The phlebothropics drugs are the therapeutic election strategy for the patients with PTS in those which is not indicated the surgery or in those which this is a assisting of the medical treatment. Finally, the deep venous surgery must be reserved for all those patients that suffer from venous insufficiency serious chronicle, with meaningful venous reflux and ambulatory venous hypertension. PMID- 14755907 TI - [Auditive and visual hallucinations secondary to tramadol administration]. PMID- 14755908 TI - [Post-trauma intercostal arteriovenous fistula in a patients with AIDS]. PMID- 14755909 TI - [Antidepressive treatment and subcapsular splenic hematoma]. PMID- 14755910 TI - [Psoas abscess following methicillin-resistant Staphylococcus aureus bacteremia associated with catheter]. PMID- 14755911 TI - [Pathologic fracture of the sacrum in an HIV patient treated with highly active antiretroviral therapy]. PMID- 14755912 TI - [Lower gastrointestinal hemorrhage associated with celecoxib]. PMID- 14755913 TI - [Nodular lesions of the lower limbs with long clinical course. Bazin's indurated erythema]. PMID- 14755914 TI - [Autoimmune hemolytic anemia as presentation form of systemic lupus erythematosus]. PMID- 14755915 TI - [Comparison of Phadiatop and skin tests in 130 patients with suspected allergic rhinitis]. AB - AIM: To establish the degree of coincidence in the results obtained from allergic skin-tests (prick-test) and the screening determination of specific IgE (Phadiatop) on a sample of patients with suspected allergic rhinitis (AR). MATERIAL AND METHODS: Prospective study of 130 patients with suspected AR, on whom we carried out prick-test and Phadiatop, comparing the results of both tests. RESULTS: The results of both tests were similar in 118 patients (89%). There were false negatives and false positives with both methods. CONCLUSIONS: The diagnosis of AR is based on a precise clinical history, complemented with prick-testing. The prick-test is considered the main test for the diagnosis of AR. In selected cases it is indicated to complete the study with the determination of serum specific IgE against the suspected allergens. The Phadiatop test is a specific IgE test for multiple allergens being its main use as a screening test in AR, mainly for outpatient cases. PMID- 14755916 TI - [Complicated sinusitis and nasal endoscopic surgery]. AB - BACKGROUND: Complicated sinusitis can result in a significant morbidity if not appropriately managed. Traditionally, surgical cases were treated by external approaches. Now a days, the introduction of endoscopic sinus surgery allows such complications to be endoscopically approached. AIM: This study was designed to evaluate our results in complicated sinusitis treated by endoscopic sinus surgery. METHODS: The study reviewed 18 patients with evidence of sinusitis complications treated with endoscopic sinus surgery between 1993 and March 2000. There were 12 orbital and 6 intracraneal complications. Six patients did need an additional surgical approach. RESULTS: Seventeen patients (94%) had a relief of their symptoms. One patient with a superior orbital fisura syndrome died a month later due to vascular and respiratory complications. Another patient with fronto lateral sinusitis required frontal osteoplasty for persistent disease. Two patients had long-term neurological sequelae (seizure). None had ocular sequelae. There were no complications due to the surgery. CONCLUSION: The endoscopic sinus surgery is a safe procedure with a high success rate in the treatment of complicated sinusitis. At present, open approaches to the sinuses are rarely indicated. PMID- 14755917 TI - [Trans-septal endoscopic approach of pituitary tumors]. AB - The advances in endoscopic instruments have eased the approach to the sellar region through the nasal cavity. We carry out an analysis of the surgical results on 20 patients that underwent surgery for sellar tumours through a transeptal transphenoidal approach in the last 2 years in our hospital. The average was 45.6 years old, and 75% were females. 30% of cases were pituitary adenomas and another 30% acromegaly, 25% Cushing's disease and 10% prolactinomas. No complications were encountered during surgery being the most common postoperative complications, diabetes insipida in two cases (10%) and CSF leak in one case. At present 2 patients are having hormonal treatment for panhypopituitarism. No patients developed a septal perforation, nasal deformity, epistaxis, meningitis, lip numbness or oronasal fistula. The rest did have good results noith no recurrence and hormonal values back to normal. PMID- 14755918 TI - [Results of supraglottic laryngeal cancer treatment with endoscopic surgery using CO2 laser]. AB - A group of 253 patients with laryngeal carcinoma was treated by endoscopic laser CO2 surgery, 50 of them presented a supraglottic localization. The follow up period ranged from 1 to 10 years. From the analysis of different parameters (age, site, stage, histopathology, etc) it could be concluded that survival and free symptoms-time rates in patients treated with laser endoscopy were similar to those obtained in patients treated with conventional external surgery. However, the endoscopic laser-CO2 surgery reported several advantages such as: no pharyngostoma, less complications, shorter stay in hospital and, consequently, lower health costs. In addition, tracheotomy was almost never necessary. Recurrencies can be also treated endoscopically, but, when total laryngectomy is needed following recurrency, this is considered to be a bad prognostic factor. PMID- 14755919 TI - [Treatment of the NO neck in supraglottic cancer]. AB - INTRODUCTION: Occult regional metastases in supraglottic cancer NO are the object of classical controversies. The aim of our study is to provide our experience in order to determine how neck treatment affects on regional recurrence of NO supraglottic cancer. METHODS: A retrospective study of 246 patients with NO supraglottic cancer treated in our service between 1977 to 1999 is presented. RESULTS: 11.4% of patients did not have any cervical treatment, with a 23% of neck recurrence amongst the evolution. 66.7% of patients were treated with a modified radical neck dissection, 24% of these presented occult metastases in the histopathological study. Global regional recurrence was 2.9% in early stages (T1/T2) and 13.1% in advanced stages (T3/T4). In pN+ patients, 85% underwent postoperative radiotherapy, with a regional recurrence of 8.8%. Postoperative clinical control without any other treatment in pN0 patients showed a neck recurrence of 6.1%. We performed a unilateral neck dissection in those patients with clear-lateral tumours. In these cases the clinical control of the contralateral neck gave a 5.5% recurrencies on that side. Patients treated with elective primary radiotherapy suffered a 5.5% of regional recurrence. DISCUSSION/CONCLUSIONS: Neck treatment of the NO supraglottic cancer is recommended. We treat neck in the same way of primary tumour (surgery or radiotherapy) with good control of regional recurrencies, less than 10%. In case of a negative pathological study of the neck careful, watching is the elective attitude. In the positive pathological study of the neck (pN+), radiotherapy is the elective treatment in those with three or more affected nodes or capsular breakdown in any of them. PMID- 14755920 TI - [Clinical course in patients treated for primary hyperparathyroidism. Our results and review of the literature]. AB - Our aim was to verify the clinical improvement following surgery in patients diagnosed of primary hyperparathyroidism. Between april 1990 and november 2002 we have operated 83 patients with this diagnosis (12 men and 71 women). Before surgery, calcium and paratohormone (PTH) levels were measured, Together with clinical symptoms. We analysed the kind of surgery, AP results, complications and clinical evolution. In over 97% of cases we obtained a biochemical normalization after surgery, 96% of the patients with nephrolithiasis improved notoriously, being this the only statistically significant symptom (p < 0.01). The rest of symptoms also decreased following parathyroidectomy (p > 0.05). PMID- 14755921 TI - [Non-benign paroxysmal positional vertigo]. AB - Positional vertigo is a frequent clinical manifestation of vertigo of very different etiologies, being the benign paroxystic positional vertigo (BPPV) the most frequent one of them, representing in some series even the most found etiology of peripheral vertigo. Usually of severe entities, positional vertigo may appear in the context of severe entities and of difficult diagnosis. In these cases, the bearing of the symptoms in spite of the repositioning manoeuvers, the association with otological or neurological symptoms, and the atypical nystagmus evoked by Dix-Hallpike manoeuver, must take the otolaryngologist to suspect of a feasible non benign pathology. We report a case of positional paroxysmal vertigo caused by an intracranial tumour and we review the clinical signs that shoved help us to suspect of non benign pathologies that can mimic a positional vertigo. PMID- 14755923 TI - [Clinic epidemiology: what for or for what?]. PMID- 14755922 TI - [Report of a case: non-Hodgkin's lymphoma of the hypopharynx in a patient with HIV infection. Remission without lymphoma-specific treatment]. AB - Non-Hodgkin's lymphomas (NHL) are common in HIV patients, although it is rare for primary lymphomas to develop in the larynx or hypopharynx. We present the case of a patient that was diagnosed of a high degree NHL, following a biopsy of the piriform sinus' lesion, taker by direct laryngoscopy. Simultaneously he was diagnosed as HIV. The interest of this paper is the total remission of the lymphoma seen after specific HIV treatment only. PMID- 14755924 TI - [SARS (Severe Acute Respiratory Syndrome). Emergent transmissible disease]. AB - Of the reemergent transmissible diseases of the past decades, SARS is probably not the last to express the alterations occurring in the relationships of the human being with its global ecosystem. The life of contemporary man is characterized, among others, by a huge thirst for traveling, for varied reasons, consequence of the globalization process. SARS virus, mutant belonging to Coronaviridae, occurred in one of the most densely populated areas of the world. There are two main moments marking the reemergence and evolution of SARS: firstly, the onset of the epidemic in China in November 2002 followed by the worldwide spread of the epidemiological process, and secondly the discovery of SARS virus as a mutant of coronaviruses in March-April 2003 in USA, Canada, and Hong Kong. The possibilities of general and special prevention, and particularly vaccine prevention are likely to bring this disease under control. PMID- 14755925 TI - [Current aspects of hemolytic uremic syndrome in children]. AB - The authors are approaching the problem of hemolytic uremic syndrome, a common cause of acute renal failure in children. This review present an update about the pathophysiology of typical hemolytic uremic syndrome, useful for understanding the clinical picture and the base for some modern therapeutical models. Concerning the evolution, the authors underline the importance of identifying the risk factors for acute phase, the extrarenal manifestations being considered of vital risk. The atypical hemolytic uremic syndrome still has some uncertainties; the article try to make a synthesis of ethiopathogeneity, clinical manifestations, evolution and therapeutical modern approaches. PMID- 14755926 TI - [Importance of genetic factors in the pathogenesis of gonadal and postgonadal human infertility]. AB - The infertility is a important health problem, affecting about 10-15% of couples. The important role of genetic factors in pathogenesis of infertility is now increasingly recognized and our knowledge in this field are improved each day. For these reasons we review the most important genetic causes of infertility. In this paper we analyse the genetic implications in gonadal and postgonadal infertility. Gonadal infertility affects both sexes and are characterised by hypergonadotrophic hypogonadism. Gonadal infertility is produced by chromosomal or monogenic mutations. Chromosomal causes are represented by gonosomal aneuploidy and structural chromosomal abnormalities. The monogenic disorders are consequences of a recessive mutations of hormone, hormonal receptor or enzymes genes. Postgonadal infertility is present in men and is the result of some obstructive disorders. PMID- 14755927 TI - [Absence seizure--recent physiopathologic data]. AB - The progresses in clarifying the normal and pathologic cellular and molecular mechanisms are reflected in the elucidation of the way some of the most common forms of generalized seizure--absence seizures--occur and are produced. Intrinsic properties of the thalamic neurons that give them the ability to release or preserve oscillatory, low-frequency neuronal discharges, and the thalamo-cortical feedback mechanism seem to explain the pathogenesis of absence seizures. The involvement of GABA receptors in the regulation of membrane calcium channels, as well as their genetically-induced changes are new pieces in the pathogenic puzzle. PMID- 14755928 TI - [Cytosolic calcium dynamics and smooth muscle contraction.III Plasmalemmal calcium fluxes]. AB - Smooth muscle contractile activity depends upon cytosolic Ca2+, the Ca(2+) sensitivity of actin-myosin interaction and several auxiliary mechanisms. This section presents the plasmalemmal Ca2+ fluxes in relation with the functional structure of their supportive proteins and the contractile impact. Summaries of classical data are accompanied by examples of recent advances. Ca2+ influx occurs mainly via the L and T types of voltage-operated Ca2+ channels, the store operated Ca2+ channels and the non-selective cation channels (operated by membrane receptors or mechanical stimuli). The plasmalemmal Ca2+ ATPase and Na/Ca antiport function to limit increases in cytosolic Ca2+; Na/Ca effect is opposite when driven to operate in the reverse mode. PMID- 14755929 TI - [HDL-cholesterol-- active or passive participant in the pathogenesis of atherosclerosis]. AB - It is known that high sanguin levels of cholesterol and LDL-cholesterol (LDLc) have an important role in the pathogenesis of atherosclerosis. The treatment of hypercholesterolemia with statins and/or with fibrates have had beneficial effects on coronary heart disease and on other localization of atherosclerosis. The decreased of cholesterol and LDL-cholesterol is the most important effect of this treatment. The epidemiological studies have revealed that the treatment with statins and/or with fibrates produce an increase of HDL-cholesterol (HDLc), which is also very important in the regression of atherosclerosis. We tried in this review to explain the mechanisms of the increase of HDL-cholesterol, in concordance with the data from literature. PMID- 14755930 TI - [Digestive implications of human immunodeficiency virus infection]. AB - AIDS, still present with severe prognosis at the beginning of the 3rd Millennium, may present gastrointestinal and hepatic involvement. Even they have not a fatal prognosis, in order to improve the quality of life of AIDS patients we need a broad knowledge of and to treat the gastrointestinal and hepatic manifestation of the disease. PMID- 14755931 TI - [Liver metastases with unknown primary site]. AB - Patients with unknown primary site cancer represents 5% to 10% of all neoplasia patients. Liver is a favourite site for gastrointestinal tumors, but not only. The adenocarcinomas represents 60% of patients from this group. A detailed physical examination, extensive laboratory and imaging procedures are necessary to locate the primary tumor. However, liver biopsy could be essential for histological diagnosis and important to identify the tumors who may benefit from specific and effective therapy (breast cancer, prostate cancer, ovarian cancer and small-cell carcinoma of the lung). Systemic chemotherapy represents the most frequently treatment, but only patients with good performance status and without co-morbidities may benefit. "Best supporting care" may be the optimal treatment for most patients, majority with poor performance status at the time of diagnosis. PMID- 14755932 TI - Image processing in myocardial SPECT acquisition. AB - Image processing for the imagistic medicine domain nuclear medicine have long time being realised through specifical protocols which rarely allowed the intervention of other informatical modalities. Processing depends first of all on the acquisition type. This paper presents the steps of one of the most complex image processing protocols, in the case of myocardial SPECT acquisition (Single Photon Emission Computed Tomography). PMID- 14755933 TI - [Chronic cough--etiological diagnosis problems]. AB - Chronic cough is a common problem in patients who visit physicians. The three most common causes of persistence cough in nonsmokers who were not taking an ACE inhibitor and who had a normal or stable chest radiograph are: postnasal drip, asthma and gastroesophageal reflux. After a viral upper respiratory infection, it takes sometimes seven weeks for bronchial airway hyperreactivity to return to normal. By using a standard protocol, 95 percent of patients with chronic cough can be managed successfully but in some cases it may take even five months or more to determine a diagnosis and effective treatment. PMID- 14755934 TI - [Running through the liver transplantation]. AB - An overview of the organization, timing and developing of the liver transplantation is difficult to be made in terms of multiple sequences and a great variety of activities during the developing of such activity. A well trained transplant team must carry out the potential donor, the liver grafts manipulation and the graft receptor, in the condition of a competitive medical system. A summary presentation, showing the essentials of the proceedings in liver transplantation could be assimilated as a guide of multidisciplinary sequences that leads to the completion of the liver grafting. The common feature of all that means the liver transplantation and generally in organ transplantation is the performance and exactingness. PMID- 14755935 TI - [Mucoviscidosis: Background, risk factors, diagnosis strategy, treatment principles]. AB - Known as a systemic disease, genetically conditioned, mucoviscidosis or cystic fibrosis brings together chronic obstructive bronchopathy, manifested since early childhood, chronic bronchial infection especially with pseudomonas or staphylococcus aureus, pancreatic impairment and a high clorum ratio in sweats. Cystic fibrosis transmembrane conductance regulator is inducing the disease, 1 of 20 caucasians being carriers of this marker. Positive diagnosis is suggested when respiratory and digestive symptoms meet fertility disfunction and "salty kiss" due to sodium--clorure disorders and confirmed by chest X-ray, spirogram, bacteriologic exam of sputum, genetic analysis and positive tests of the pancreatic enzymes and sweat's test. As regards therapeutical solutions, besides antibiotics required in infectious exacerbation, corticotherapy, kinesitherapy and alpha dornasis given in aerosols to improve bronchial mucosa's clearance, also substitutes of pancreatic enzymes and hypercaloric diet are to be considered. In the last two decades, lung transplant appeared to be as an alternative to improve poor prognosis of this disease. Recent studies reported that life expectancy in cystic fibrosis will increase from 28 years in the '90s to 40 years in the next 20 years. PMID- 14755936 TI - [The neurotransmitters and alcohol dependence]. AB - Exist many interactions of the endogenous system with dopaminergic, serotinergic, GABA--ergig, and glutamat--ergic, as well as the role of these interactions in mediating stress response, analgesia and in appearance the alcoholism. Thus pharmacological blockade of the endogenous opioid system by mu or delta opioid receptor antagonists prevents ethanol's activation of the dopamine system and reduces ethanol consumption. The role of genetic factors in alcohol dependency is recognized. The applicability of PET, SPECT, MRS, functional neuroimaging in the study have a very importance, for demonstrates the changes in regional cerebral blood flow at the alcoholics. PMID- 14755938 TI - [Cyclodextrins implications in drugs control and analysis]. AB - The discovery and development of substances require new forms at a level which was considered satisfactory a few years ago. The research on secondary effects and drugs optimizations to new pharmaceutical forms. So, we can speak about the inclusion complexes. The purpose of the paper was to describe the cyclodextrins properties, the complexation process and the most important analysis methods for complexes between cyclodextrins and drugs. PMID- 14755937 TI - [Oral disintegrating tablets. A new, modern, solid dosage form]. AB - The pharmaceutical market shows lately an increasing interest in orally disintegrating tablets, due to their good acceptability among certain age categories (ex. elderly, children), and other patients with difficulties in swallowing classic solid dosage forms. Some of the methods of preparing such tablets have gained industrial applicability: molding, lyophilization, direct compression with highly soluble excipients, super disintegrants and/or effervescent systems. Some of the patients have had a good impact on the pharmaceutical market and more improvements are expected in the next few years, with new drugs to be formulated as fast dissolving dosage formulations. PMID- 14755939 TI - [Overexpression of c-erbB-2 gene product is associated with poor prognosis factors in breast carcinoma]. AB - Oncogenes, the abnormal forms of proto-oncogenes, were shown to be involved in malignant transformation and in tumor progression. c-erbB2/HER2/neu is member of EGFR family and encodes the p185 protein, which functions as a tyrosine-kinase. Gene amplification and/or p185 overexpression were reported to be associated with poor prognostic in cancer. Our purpose was to investigate p185 immunohistochemical expression in breast carcinomas and in the corresponding axillary lymph nodes metastases and to identify possible correlation between p185 and other factors of poor prognostic, such as loss of hormonal receptors expression. In our study, 40.91% of cases were erbB-2 positive, p185 expression being maintained from the primary tumors to axillary metastases and associated with positive nodal status and with the absence of hormonal receptors expression (p < 0.05). These findings support the hypothesis the c-erbB2 is an advantageous acquisition for the aggressive behavior of the tumor cell and for its ability to invade and metastasize. PMID- 14755940 TI - [Statistical and regression analysis in interpreting computerized morphometric results]. AB - The aim of the study is to construct the statistical framework requested by the interpretation of the results obtained through computerized morphometry procedures. The study also includes the regression analysis of the investigated quantitative features. MATERIAL AND METHODS: The automatic measurements performed under Zeiss KS400 environment were achieved on a microscopic specimen of dental pulp. The areas and the perimeters were determined for 139 fibroblasts, so as to define two statistical series able to characterize the size of the cells. RESULTS AND DISCUSSIONS: The results are formulated in both analytic and numerical terms, the latter relying on the graphical support and computational resources of Zeiss KS400 environment. Also, the mathematical support is implemented. The research focuses on three key aspects: (i) the role of the class organization (clustering) in the correct evaluation of the measured cellular areas, (ii) the global characterization of these areas by the standard statistical indices, (iii) the linear and quadratic regression perimeter versus area. CONCLUSIONS: The study is based on a concrete morphometric investigation of a dental pulp microscopic specimen, but the statistical and regression analysis can be applied to any other kind of specimen that undergoes a morphometric examination. The implementation under Zeiss KS400 is easy adaptable to other software environments providing similar facilities. For an exhaustive approach, in accordance with the sampling and selection laws, this type of study should be successively practiced on several fields of the microscopic specimens. PMID- 14755941 TI - [Polyamines antagonizing angiotensin II contractile effects in isolated rat aorta]. AB - Our study showed that the administration in pre-treatment of some polyamines (especially spermine and spermidine and almost null agmatine, putrescine and cadaverine) reduced the contractile effects of angiotensin II (Ang II) in isolated rat aorta. These effects might not be associated to the interference of clathrin coated vesicles (coated pits) formation or caveolae interaction (and thus to Ang II internalization through AT1 receptors). In contrast, these effects seem to be due to the interaction with voltage-gated membrane Ca2+ channels. Therefore, the alteration of transmembrane Ca2+ fluxes does not exclude the involvement of internalization process through coated pits or caveolae, since the endocytosis mediated by these phenomena essentially needs Ca2+. In addition, the inhibitory effects are dependent on the number of positive charges of the polyamine molecules. PMID- 14755942 TI - [Endometrial hyperplasia predicts a better prognosis in endometrial carcinoma]. AB - AIM: To evaluate the significance of the associated Endometrial Hyperplasia in Endometrial Carcinoma. STUDY DESIGN: 93 hysterectomy specimens for endometrial carcinoma were histologically reviewed on paraffin sections stained H&E and tricromic VG. RESULTS: 45 cases were associated with endometrial hyperplasia; 48 cases without endometrial hyperplasia were: 35 cases with the entire endometrial cavity occupied by carcinoma, 11 associated with endometrial atrophy and 2 with normal proliferative endometrium. In this second group, 37 cases (77%) were in advanced stages. A correlation was found between the presence of Endometrial Hyperplasia and the degree of differentiation, the FIGO stage of the carcinoma, and the age of the patient. CONCLUSIONS: Endometrial Carcinomas associated with Endometrial Hyperplasia involve younger women and have lower aggressivity than those without hyperplasia. Endometrial Hyperplasia may demonstrate a favorable prognosis in Endometrial Carcinomas. PMID- 14755943 TI - [Practical aspects in diagnosis and treatment of benign breast lesions]. AB - We analyzed 444 patients operated on in our clinic. Triple test diagnosis is the modern trend. Surgical treatment is not justified in all cases. When needed, partial mastectomy is commonly used. Surgical principles must be respected for good cosmetic results. Histopathology only certifies the diagnostic. PMID- 14755944 TI - [Malignancy risk in some benign mammary lesions]. AB - We analyzed retrospectively 821 patients operated on in our clinic for benign mammary lesions and for breast cancer. Histopathology identified in some cases associated benign and malign lesions. Atypical hyperplasia is considered significant risk factor for malignancy. Women in this category need close surveillance. PMID- 14755945 TI - [Morphological correlations between endometrial hyperplasias, uterine leiomyoma and ovarian associated lesions]. AB - Morphological evaluation and correlation of endometrial hyperplasias, leiomyoma and ovarian estrogen producing lesions. 390 specimens of hysterectomy associated or not with ovarectomy were processed by routine technique and sections stained by H&E staining. 316 cases presented different degrees of endometrial hyperplasias associated with leiomyoma. In 117 from 170 cases the ovary presented estrogen secreting lesions (follicular cyst, polycystic ovary, stromal hyperplasia, granulosa cell tumours). The highest frequency of the uterine and ovarian lesions was in the decades 41-50 years, and 31-40 years. Leiomyoma and endometrial hyperplasia, develop in a hormonal context, most frequently characterized by micropolicystic ovaries. The risk is higher in the perimenopausal period. The most frequent type is simple hyperplasia suggesting a rare progression to highest grades and a possible protective role of leiomyoma as target tissue which capture estrogens. PMID- 14755946 TI - [Evaluation of the systolic and diastolic function of the left ventricle in patients with diabetes mellitus and microalbuminuria]. AB - The objective of this study is to reveal the deterioration of the systolic and diastolic function of the left ventricle (LV) in patients with diabetes mellitus (DM), in correlation with type of DM, the edge and the complications of DM and the associated diagnosis. METHODS: The study included 43 patients with DM (29 patients with DM type 1 or 2 insulinodependent--DMID and 14 patients with DM type 2 nonisulinodependent--DMNID). The age of patients was between 21 years and 57 years, with a duration of DM between 1 year and 22 years. The ecocardiographic evaluation have determined 38 parameters (B mod, M mod, CWD, PWD, colour Doppler). From these parameters, 12 was used for the measurement of the systolic function of the LV and 18 was used for the determination of the diastolic function of the left ventricle. The clinical and biological exam have performed: glycaemia, proteinuria, dyslipidemic syndrome, complications of DM and ischaemic heart disease (ECG). RESULTS: The ejection fraction (EF) of the LV was ?60% in 23 patients, between 50% and 60% in 17 patients and between 40% and 50% in 3 patients. The values of MSER (mean systolic ejection rate) was between 221 ml/s si 515 ml/s and the values of MVCF (mean velocity of circumferential shortening) was between 0.9 circ/s si 1, 8 circ/s. The ratio of the E and A waves at the mitral valve was > 1 in 22 patients and < 1 in 21 patients. The LV isovolumic relaxation time (IVRT) was < 70 ms in 13 patients, > 100 ms in 14 patients and had normal values in 16 patients. The E wave deceleration time (EDT) was < 150 ms in 24 patients and had normal values in 19 patients. There is no connection between the systolic/diastolic function and the other determined parameters. CONCLUSIONS: The alteration of the LV systolic function was present in 20 patients from 43 patients. The LV diastolic dysfunction through relaxation troubles was present in 14 patients and the LV diastolic dysfunction through compliance trouble was present in 13 patients. A number of 16 patients had preserved (for the moment) a normal LV diastolic function. The number of patients with these two types of LV diastolic dysfunction was equal in our study. Other studies had found a predominance of LV diastolic dysfunction through compliance troubles in DM. PMID- 14755947 TI - The role of some prostaglandin analogues in experimental intoxication produced by carbon tetrachloride in rats. AB - Prostaglandins are synthesized ubiquitously in the body from unsaturated fatty acids and they act as paracrine messengers. We have studied the influence of a prostaglandin analogue on experimental induced hepatopathy. The tested compound is a synthetic isopropyl ester of PGF2 alpha (IPEF) and as hepato-toxic agent we used CCl4. We worked on four groups of 4 adult male rats each. Group I received no substance; Group II received CCl4 0.1 ml/bw/per os, single dose, for three days; Group III received CCl4 as series I and IPEF 15 micrograms/bw i.p., single dose daily, one hour before CCl4 administration; Group IV received CCl4 as series I and IPEF 50 micrograms/bw i.p., single dose daily. Twenty-four hours after the last administration, samples of blood were taken and ALT, AST, LDH as well as conjugate and unconjugate bilirubin were determined. We also determined MDH, GSH and glutathion peroxidase, in liver homogenate. Our data show that MDH levels are increased in Group I (20.81 +/- 3.15 microM/microgram protein) as compared with both Group III (8.44 +/- 1.32 microM/microgram protein) and IV (7.31 +/- 1.92 microM/microgram protein) which might suggest that prostaglandin analogue IPEF decreases the polyunsaturated fatty acids degradation, at both low and high level. ALAT levels for group that received CCl4 (782 +/- 20.8 U/L) are significantly higher than those for group III (264 +/- 15.4 U/L) and IV (227 +/- 8.4 U/L) which received IPEF at low, respectively high dose. Our data suggest that the synthetic prostaglandin analogue presents stabilizing membrane effects (plasmatic and membrane of some cellular organelles) and reduces peroxide radicals production. PMID- 14755948 TI - [Clinical aspects of hypertension in children with aortic coarctation]. AB - Our study is focused on blood pressure before and after repair of aortic coarctation in childhood. METHODS: A group of 26 children (13 boys, 13 girls, range 1-18 years, of which 12 operated: 9 boys and 3 girls) was studied, blood pressure being followed before and after operation. The recorded blood pressure was compared to normal values for age and height. RESULTS: Before the surgical treatment blood pressure being above normal in all cases, the figures were: "high normal"--6 cases, significant--8 cases and severe hypertension--12 cases. After surgery: 75% normal blood pressure, 25% hypertension (variable degrees). CONCLUSION: Hypertension in aortic coarctation varies from "high normal" to severe. Hypertension got worse during pregnancy in an unoperated girl. In most of children, blood pressure decreased after surgical treatment, being normal in 75% of all cases, in one year after surgery. Persistence of a severe hypertension after surgery signifies presence of an underlying lesion unrecognised yet. PMID- 14755949 TI - [Some aspects regarding degenerative mitral valvular lesions encountered in medical practice]. AB - In the last years, the degenerative valvular heart diseases have the tendency to equalize in frequency the rheumatismal valvular diseases. The maximum attention has been paid on the degenerative aortic stenosis as being a lesion with maximum frequency and a severe evolution. This study, given on the 18391 admissions in the period 1997-2001, is a retrospective analyse and it is concerned with the degenerative mitral valvular lesions. Of the 223 patients with degenerative valvular heart lesions, 139 patients (62.3%) had degenerative aortic stenosis and 96 patients (38.5%) were diagnosed with degenerative mitral valvular lesions from which 30 patients have had no association with aortic valvular lesions while 66 patients have had such an association. The pointed out types of mitral lesions were: the mitral insufficiency in 59 patients, the mitral annular calcification without hemodynamic disease in 19 patients, the mitral stenosis in 9 patients and the mitral disease in 9 patients, too. The women was affected nearly 1.7 times more frequent than the men, with a maximum average age greater with four years for women but with a low minimal average age at 60 years for women and 52 years for men. The detailed analyse of this 96 cases had shown the presence of a cholesterol value over 200 mg/dl in 50 patients (52%), the diabetic mellitus of type II in 12 patients (12.5%), an association with HTA in 42 patients (43.7%), the cardiac insufficiency in 68 patients (70.8%), a permanent atrial fibrillation in 24 patients (25%), the chronical myocardiac infarct in 19 patients (19.7%) and disorders in the transmission of stimuli in 8 patients (8.3%). The degenerative mitral valvular lesions had occurred more and more frequently realizing more complex features under the mitral insufficiency predominance. Its frequent association with the degenerative valvular lesions determines the evolutive and therapeutic particularities that are dominated by the high gravity prognostic. PMID- 14755950 TI - [Ureterolithotomy versus retrograde ureteroscopy for lumbar ureteral stone. A cost analysis]. AB - Ureterolithotomy is considered to be the last option for treating a lumbar ureteric stone. The authors reviewed two cases with lumbar ureteric stones treated in january 2003, one by ureterolithotomy and one by retrograde ureteroscopy regarding the costs for hospitalisation, investigations and medication. Among the certain clinical advantages for the patients, including a smaller rate of complications, ureteroscopy is considerably cheaper than open surgery. We consider that these costs criteria should be taken into consideration when the necessary equipment for-urological departments are set. PMID- 14755951 TI - Complications following gastric tube esophagoplasty in children. AB - Surgery in children with caustic esophageal burns, esophageal atresia or other esophageal disorders can be performed using gastric tube esophagoplasty. Between 1991 and 1999, a number of 41 such procedures have been performed in our department, using the original technique developed by Gavriliu. The results were assessed as good and very good in 83% of the cases, and fair or bad in the rest of 17%. We recorded 13 cases with significant complications, 4 of which ended with exitus. The paper present our experience in using gastric tube esophagoplasty and the management of the complications related to this procedure. PMID- 14755952 TI - [Dynamic condylar screw in reverse oblique trochanteric fractures]. AB - The aim of the study was to evaluate the results of internal fixation with Dynamic Condylar Screw (DCS) in reverse oblique trochanteric fractures. In a two years period 238 patients with trochanteric fractures were treated in our department. In 31 cases (13%) the fracture has reverse obliquity (type II Evans). Internal fixation was performed with Ender nails (4 cases), AO condylar blade plate (11 cases), Dynamic Hip Screw (3 cases), Gamma nail (3 cases) and DCS (10 cases). Results 4 month after the surgery reveal that Gamma nail and DCS used in reverse oblique fractures were superior to other types of implants. Placement of DCS may be difficult in fractures with large displacement and technical solution in such fractures are presented. In conclusion we consider that, in type II Evans trochanteric fractures, internal fixation with DCS is a good option. PMID- 14755953 TI - [Prevalence of type 2 diabetes mellitus in an internal medicine ambulatory care department]. AB - The aim of the study was to evaluate Glico regulation disturbances prevalence (glycemia over 126 mg/dl). We analyzed, in a retrospective study, 3051 patients hospitalized in Internal Medicine Ambulatory Care Department, "St. Spiridon" Hospital, during a period of two years, 2000-2001. Our department was design for one-week spitalization for diagnostic, so the patients are not on emergency condition. The medium age was 53 +/- 3 years, 24% males, and 76% females. We were looking for clinical and biological criteria of diabetes and co-morbidities (hypertension, dyslipidemias, ischaemic heart diseases). We also studied symptoms of admission and complications as infections. We found a jeune glycemia l126 mg/dl on 151 cases, 4.8% of total cases, predominantly women, 88 cases (58.3%), 63 males (41.7%). 44 cases were less than 50 years old and the rest over, 107 cases (70%). We excluded type 1 diabetes mellitus already known. The evaluation of BMI shows us the predominance of overweight and obesity (83.44%), 126 cases. 125 cases had cholesterol level over 200 mg/dl, 82.78%, and only 26 cases had lower. From these, 32% had triglycerides > 150 mg/dl. Study of co-morbidities revealed 42 cases with normal arterial tension, the rest had arterial hypertension, ischaemic heart diseases, 109 cases, 72.18%. More than a half had infections, especially urinary and pulmonary. Admission had relevant accuses: 34.27% came for diabetes suspicions, 28.31% had cardiovascular diseases, 12.36% had symptoms which were relevant for diabetes. We concluded that from the group of patients that are searching for diagnostic, on an internal medicine department, 4.8% had type 2 diabetes. We underline strong association with dyslipidemia and obesity. PMID- 14755954 TI - Multi-valued logic in breast cancer detection. AB - The aim of this paper is to determine the type of the breast cancer disease. The two classes of separation are malignant respectively benign. A multi-valued logic system (fuzzy system) was develop and applied in this classification. The system uses nine attributes as inputs that were scaled with an integer value in the range between 1 and 10. The attributes represent: 1. Clump Thickness, 2. Uniformity of Cell Size, 3. Uniformity of Cell Shape, 4. Marginal Adhesion, 5. Single Epithelial Cell Size, 6. Bare Nuclei, 7. Bland Chromatin, 8. Normal Nucleoli, 9. Mitoses. After training the system managed to get a good detection with an error less than 5%. PMID- 14755955 TI - [Alcohol intake assessment in a group of hypertensive adults]. AB - The assessment was realised in order to obtain data for a local intervention program for preventing primary hypertension in young adults through an education program. 269 persons, of both sexes, were investigated. The frequency of elevated blood pressure values is 11.2% and the prevalence of arterial hypertension is 8.73%. The frequency of alcohol consumers is different according to gender (18.6% for male and 3.4% for female). PMID- 14755956 TI - [VIS spectrophotometry used for quantitative determination of iodide in food products of animal origin]. AB - The spectrophotometric method used in dozing the iodides in the water, has been studied in order to use it in determining the iodides in a series of food products. This method is based on the catalyzed reduction of the iodide of Ce4+ by the arsenious acid. In order to determine the iodide in several food products of animal origin (eggs, milk and meat), the sample is first treated with a concentrated KOH solution and is maintained on a water bath until is obtained a residue; this is then subjected to calcination at temperatures of 500-600 degrees C until a perfectly white ash is obtained. After cooling, the ash is dissolved in water, concentrated H2SO4 is added and is completed with water up to a certain volume. The thus obtained solution is used for determination of the iodide in food, according to the method described above. The content of iodide (microgram%) have been calculated with the relation deduced from the equation of the regression line delineated in VIS at 459 nm. The method is selective and the procedure is widely applicable to the determination of iodide in different food products of animal origin. PMID- 14755957 TI - [Researches on methyl-xanthine series.XI. Synthesis and physicochemical characterization of some 7-R-1,3-dimethyl-xanthine derivatives]. AB - We have synthesized new xanthine compounds, derivatives of theophylline with various radicals in 7 position. The compounds have been characterized by molecular formula, weight, yield and melting point. The synthesized xanthine derivatives have been purified by repeated crystallizations from various organic solvents. Their chemical structure was confirmed by IR spectroscopy. PMID- 14755958 TI - [Distance learning using internet in the field of bioengineering]. AB - The Leonardo da Vinci training programme supports innovative transnational initiatives for promoting the knowledge, aptitudes and skills necessary for successful integration into working life. Biomedical engineering is an emerging interdisciplinary field that contributes to understand, define and solve problems in biomedical technology within industrial and health service contexts. Paper presents a Leonardo da Vinci pilot-project called Web-based learning and training in the field of biomedical and design engineering (WEBD). This project has started on 2001. The WEBD project proposes to use advanced learning technologies to provide education in the www. Project uses interactive 3D graphics and virtual reality tools. The WEBD distance training permits users to experience and interact with a life-like model or environment, in safety and at convenient times, while providing a degree of control over the simulation that is usually not possible in the real-life situation. PMID- 14755959 TI - [When is it a diagnosis of overlap syndrome?]. AB - The connective tissue diseases comprise a group of syndromes of unknown etiology affecting as many as 1 person in 40, often with a predilection for the female sex. Included are: systemic lupus erytematosus (SLE), polymyositis and dermatomyositis, Sjogren syndrome, scleroderma and the vasculitis (polyarteritis nodosa, Wegener's, giant cell arteritis). There are patients who are not easily defined; having features overlapping with those of other connective tissue diseases. A variety of terms such as mixed connective tissue disease, undifferentiated connective tissue syndrome and overlap syndrome have emerged to describe such patients. Although many of these overlap syndromes are unlikely to have life-threatening consequences, they may be extremely debilitating and distressing, significantly reducing quality of life for the patient and his or her family. We present the case of a patient initially diagnosed with dermatomyositis and who eventually evolved to overlap syndrome by developing SLE. PMID- 14755960 TI - Cardiac leiomyosarcoma. Case report. AB - A case of primary cardiac leiomyosarcoma diagnosed at Iasi Cardiology Center is presented. The study was made on the surgical biopsy specimen, and the diagnosis by routine morphological techniques. The gross examination revealed a large intracavitary left atrial tumor attached by a small sessile base to the left posterior atrial wall, having a smooth, white-gray surface, and a dense consistency. On the cut surface, the tumor had a whirled white appearance, with focal brown areas. The microscopic examination revealed the presence of a spindle cell tumor, forming fascicles orientated at right angles. The study revealed the morphological aspect characteristic to leiomyosarcoma. PMID- 14755961 TI - [Therapeutical options in partial infected aorto- bifemoral prosthetic grafts]. AB - PURPOSE: To asses the use of synthetic or autologous conduits for the reconstruction of iliofemural sector in the cases of partial, limited infections of aortobifemural prosthetic grafts. METHOD: We present two cases, both of them with infection recognized at the groin; the first one by a vascular fistula bleeding intermittently, the second by a draining groin sinus with fever and leukocytosis. RESULTS: The PTFE conduit was the material utilized at the first case and the superficial femoral vein in the second case. The patients presented completed resolution of the infectious signs and symptoms and the revascularisation of the legs was successfully done. Evan with an extra anatomical bridge or an autogenous material, sometimes is possible to resolve a regional infection of the aortobifemural graft only with a partial resection of one leg of the prosthesis and replacing with another sterile material. PMID- 14755962 TI - [Professor dr.doc Lazar Wasserman. A pioneer of modern morphopathology]. PMID- 14755963 TI - [A masterpiece (spiritual reform) of professor Grigore T.Popa]. PMID- 14755964 TI - Microscopic morphometry--a modern approach. AB - OBJECTIVES: The aim of this paper is to give a brief presentation of the recent trends in histologic and histopathologic computerized morphometry, based on the personal experience in exploiting modern software environments and illustrated by a relevant case study. Through its goal, overall conception and direct involvement in diagnosis, the work belongs to the currently emerging researches on analytical and quantitative histology. MATERIAL AND METHODS: The proposed methodology for the automatic extraction of quantitative features is applied to an endometrial adenocarcinoma and is implemented as a procedure, called ADK, running on ZeissKS400 platform. The description of the ADK procedure, as a detailed flowchart, is preceded by an overview of the KS400 resources for biomedical domain, compared to the main characteristics of other three well-known software packages for digital image processing. RESULTS AND DISCUSSIONS: The results are formulated in terms of the numerical information provided by the exploitation of ADK procedure, referring to the 2-D and 3-D (stereological) evaluation of tumorous versus stromal areas and volumes, respectively. Procedure ADK also performed measurements for the nuclei corresponding to the stromal and tumorous zones, respectively, in order to calculate the percentage of nuclei inside these zones. The discussions focus on the interpretation of the measurement results, as factors for the prognostic assessment. CONCLUSIONS: The computer-aided morphometric investigations open perspectives for operating with more expressive criteria in pathology diagnosis, resulting from the possibility to quantify, at the pixel level, features of interest which in classical approaches are subject to rough approximations. PMID- 14755965 TI - [Chronic hepatitis B virus infection]. AB - Chronic hepatitis B virus infection affects approximately 350 million people worldwide and 1.1 million in Romania. Despite the tremendous progress in the management of virus B infection, the treatment of chronic hepatitis B remain difficult and often disappointing. The aim of treatment is sustained suppression of HBV replication and remission of liver disease. Treatment is not indicated in immune tolerance phase or in the inactive carrier state, but it is necessary in immune clearance phase, after 3-6 months of surveillance. Two antiviral drugs are accepted: alpha-interferon and lamivudine. The results of both treatments are far of being perfect; new, more potent antiviral drugs are required. PMID- 14755966 TI - [ The importance of genetic factors in pathogenesis of central infertility (hypothalamic and pituitary]. AB - The infertility is a important health problem, affecting about 10-15% of couples. The important role of genetic factors in pathogenesis of infertility is now increasingly recognized and our knowledges in this field are improved each day. For these reasons we review the most important genetic causes of infertility. In this paper we analyse the genetic implications in central infertility (hypothalamic and pituitary). These conditions affect both sexes and are characterised by hypogonadotrophic hypogonadism. In majority of cases central infertility is produced by recessive mutations of hormone or hormonal receptor genes. In some cases the infertility is a component of a specific syndrome. PMID- 14755967 TI - [Myocardial remodeling]. AB - Chamber remodeling and myocardial systolic dysfunction play an important role in the development of cardiac failure after myocardial infarction. The process is accompanied by qualitative and quantitative changes in the expression and reexpression of genes regulating cell growth and contractile function. The article tries to explain some of the mechanisms of the myocardial remodeling that permit the modifications and adaptation of the heart to normal and pathological conditions from a clinical point of view and, in the same time, from a molecular point of view. PMID- 14755968 TI - [Antisense oligonucleotides with pharmacologic action]. AB - The potential therapeutic applications of the antisense strategy are illustrated by numerous examples of the oligonucleotides investigated in preclinical and clinical trials especially for antiviral, antiinflammatory, anticancer and antiatherosclerotic activity. The main advantages of the antisense oligonucleotides therapeutic candidates are comparatively discussed with the classical drugs. PMID- 14755969 TI - [Endocrine-metabolic disturbances in chronic stress]. AB - This review focuses on the neuroendocrine infrastructure of the adaptive response to chronic stress and on its concerted effects on behavior, the major endocrine axes and immune systems. Cytokines such as TNF-alpha, IL-1, and IL-6 and other humoral mediators of inflammation are potent activators of central stress responsive neurotransmitter systems, constituting the afferent limb of a feedback loop from the immune/inflammatory system to the central nervous system. Steroid hormones influence a variety of neuroendocrine events, including brain development, sexual differentiation and reproduction. Hormones elicit many of these effects by binding to neuronal steroid receptors, which are members of a nuclear receptor superfamily of transcriptional activators. Nuclear receptor coactivators enhance the transcriptional activity of steroid receptors. PMID- 14755970 TI - [Nonalcoholic and alcoholic steatohepatitis]. AB - NASH (Nonalcoholic Steatohepatitis) and ASH (Alcoholic Steatohepatitis) are both diseases with increase frequency which may complicate up to 50% of liver cirrhosis. They are considered as risk factors for liver carcinoma. In order to diminish the incidence of liver cancer, NASH and ASH need a close follow-up. These two entities need to be known not only by gastroenterologist, but also by general practitioners, endocrinologists, nutritionists and cardiologists. PMID- 14755971 TI - Helicobacter pylori infection in patients with diabetes mellitus. AB - Data concerning the prevalence of Helicobacter pylori (H. pylori) in diabetic patients are scanty and controversial. The aim of this study was to evaluate the prevalence of H. pylori infection in patients with diabetes mellitus (DM), and to assess whether the presence of bacterium was associated with severity of dyspeptic symptoms and endoscopic findings in such patients. The study involved 42 patients (19 men, 23 women; mean age 55 years, range 34-75 years) with DM and dyspeptic symptoms. Sixteen patients (38%) were classified as having type 1 diabetes and 26 (62%) patients as having type 2 diabetes. All patients had chronic dyspepsia, and each patient has completed a self-report questionnaire to obtain information concerning the presence and severity of upper gastrointestinal tract symptoms. H. pylori status was confirmed by 13C-urea breath test (13C-UBT), and diabetic patients underwent upper gastrointestinal endoscopy. Twenty-six (61.9%) of patients with DM were positive for 13C-UBT. There were no statistically significant differences in the infection rate between patients with type 1 and type 2 diabetes, and the prevalence of H. pylori infection was not associated with the known duration of diabetes. There was no significant difference in the symptoms score between H. pylori-positive and H. pylori negative diabetic patients, and endoscopic findings in patients with DM were in the same range with those found in dyspeptic subjects from the same region. In conclusion, H. pylori infection is not associated with DM, duration of diabetes, or severity of dyspeptic symptoms in patients with DM. PMID- 14755972 TI - Evaluation of simvastatin antioxidant effects. AB - 3-Hydroxy-3-methyl-coenzyme A reductase (HMG-CoA reductase) which transforms 3 hydroxy-3-methylglutaril-coenzyme A (HMG-CoA) in mevalonate, is the rate limiting enzyme in cholesterol biosynthesis. In our study, from HMG-CoA reductase's known inhibitors, we used simvastatin (ZOCOR), which is a semi synthetic derivative of the second generation. The study was performed on 25 subjects (12 men and 12 women) aged 33-67 yo, with hypercholesterolemia, which have received simvastatin, 10 mg daily for 8 months. Under treatment with simvastatin we obtained a significant decrease of total cholesterol (p < 0.0001) and an improvement of enzymatic antioxidant parameters: superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase (CAT). In conclusion, simvastatin therapy determines a significant decrease in SOD, GPx, CAT, and an increase in CAT/SOD and GPx/SOD ratios. PMID- 14755973 TI - [Clinico-functional particularities of cardiac dysfunction in the elderly]. AB - Ischaemic left ventricular dysfunction is more frequent at the elderly and has a series of clinical and functional peculiarities. The aim of the paper is to define the clinical, biochemical, and echocardiograph features of the older patients with ischaemic heart disease. METHODS: 189 patients with ischaemic heart disease (old myocardial infarction, angina, ischaemic cardiomiopathy) have been parted, according to age, in Group A (n = 101) over 60 years old and Group B (n = 88) < 60 years old. Symptoms, cardiovascular risk factors, lipid profile, echocardiograph findings, and the exercise testing (at the bicycle) were analyzed. RESULTS: Ischaemic heart dysfunction appears earlier at the male gender, who is dominant at Group B (83%) in comparison with Group A (48%, p = 0.009). Myocardial infarction is more frequent at Group B (61 vs 42%, p = 0.006), and ischaemic cardiomiopathy at Group A (34% vs. 2%, p = 0.004). At Group A, symptomatic heart failure (dyspnoea of III or IV NYHA class) is prevalent (20% vs. 1%, p = 0.002). Arterial hypertension was dominant at Group A (73% vs. 58%, p = 0.02). At the echocardiograph examination, regional and diffuse contractility abnormalities were dominant at Group A, and the ejection fraction was lower (49 +/- 10%) in comparison to Group B (53 +/- 9%, p = 0.005). Diastolic dysfunction was found in 63% at Group A and 36% at Group B (p = 0.001). At the exercise testing there were not reported significant differences concerning the mechanical load according to age (87% of maximum heart rate at Group A and 83% at Group B). CONCLUSIONS: At the old patients with ischaemic left ventricular dysfunction the presence of a myocardial infarction is not so frequent. Systolic and diastolic dysfunction of the left ventricle is more severe, the ejection fraction is lower, but the exercise capacity did not differ significant. PMID- 14755974 TI - [Abnormal positions of the heart. An analysis on 1039 cases]. AB - The aim was to study the impact of the cardiac malpositions into the group of the 1039 congenital heart diseases registered in the Pediatric Outpatient Department of "Sf. Spiridon" hospital. All patients were investigated noninvasively using clinical examination, electrocardiogram, routine Roentgenogram, two-dimensional and Doppler echocardiography, abdominal echography and, only in two cases, cardiac catheterisation. 23 (2.21%) from 1039 congenital heart diseases registered were cardiac malpositions: dextroposition--3 cases (13.04%), dextrocardia--7 cases (30.43%) and situs inversus--13 cases (56.52%). Most of the children were boys (65.21%), 70% from all cases coming from urban area. Only 3 children had structural cardiac anomalies: two cases with dextrocardia (one with atrial septal defect and one with atrioventricular canal) and one with situs inversus and tetralogy of Fallot, two of them suffering surgical correction. Psychological impact was the main problem of these children, especially during the adolescence, except the two cases with structural cardiac abnormalities who needed following and surgical treatment. PMID- 14755975 TI - [Aspects of left ventricular function in children with chronic renal failure]. AB - OBJECTIVE: To present the main morphological and functional cardiac modifications in children with chronic renal failure (CRF) and the effect of hemodialysis on systolic function of the left ventricle (LV). METHODS: 22 patients (5-19 years) with CRF of different degrees of severity. 16 of them were included in a hemodialysis (HD) program and they were followed-up for at least 1 year. The systolic and diastolic function of the LV were investigated by echocardiography (echo) 2D and Doppler. RESULTS: Although the clinical signs of cardiac suffering were rare, echo detected concentric hypertrophy (9 cases), dilated cardiomyopathy (4 cases), mitral and tricuspidian regurgitation and pulmonary hypertension (3 cases) and pericardial effusion (10 cases). Systolic function was normal in most of the cases, but the diastolic function (evaluated by E/A ratio, isovolumic relaxation and deceleration time) was modified in 10 cases. The HD session induced the improvement of the LV systolic function parameters and the decrease of right ventricle dimensions. CONCLUSIONS: Echocardiography has a higher sensitivity compared to ECG and chest X-ray in evaluation of the LV function, mainly in the cases without cardiac symptoms. An efficient chronic HD program and also a complex antihypertensive treatment contribute to the alleviation of the LV function. For the HD session, echo is useful in monitoring the systolic LV function. PMID- 14755976 TI - What added value does ambulatory blood pressure monitoring brings to the management of post renal transplantation hypertension? AB - OBJECTIVE: To evaluate BP control, white coat hypertension (WCH) and abnormal circadian variability in a significant outpatient sample of renal transplant (RTx) subjects, normotensive at the last regular visit. METHODS: ABPM (Spacelab 90217) was performed every 15 min between 07:00-23:00 h and every 30 min between 23:01-06.59 h. in all patients (N = 68, 39M, S-Cr. = 153 +/- 49 mumol/l) normotensive at their last regular office BP (O-BP) measurement and with available BP records for the 12 months preceding RTx and 6 months preceding ABPM. RESULTS: BP values were frequently abnormal in this RTx cohort considered to have a satisfactory BP control. O-BP (measured with a Hawksley random-0 sphygmomanometer on the day of ABPM) was 135.5/80.6 mmHg, 47.1% of the patients with abnormal BP values. By comparison, ABPM showed a lower prevalence of uncontrolled BP: 44.1% for 24 h.-BP and only 35.3% for the daytime awake period, with values of 134.5/80.4 and 135.3/81 mmHg respectively (P = NS from O-BP). The prevalence of WCH was 12%. 24-h SBP is related to O-SBP (r = 0.71, P < 0.01) and Bland-Altman analysis demonstrates that > 95.6% of all differences between systolic ABPM and O-BP values are within +/- 2SD of the identity line. However, although 24-h DBP is equally related to O-DBP (r = 0.64, P < 0.01), on Bland Altman analysis, 8.8% of the differences between diastolic ABPM and O-BP values are outside +/- 2SD of the identity line. Thus, systolic but not diastolic O-BP correlates with, and can be substituted to ABPM derived values. Non-dipping was frequent, regardless of the definition of normal nocturnal BP fall (10 mmHg or 10% of the daytime SBP): 82.4%, 89.7%. Even if normality was strictly defined as a night/day ratio < 0.90 for SBP and < 0.92 for DBP, non-dipping prevalence was high 73.5%, with more than one-third of the RTx patients having nocturnal hypertension (ratio > 1). CONCLUSIONS: BP control is not optimal in one-third of a typical RTx population. Furthermore, nocturnal hypertension is a frequent and underestimated phenomenon in this population. There is a good agreement between ABPM derived and casual systolic values. Office measurements, due to WCH, are under-evaluating the quality and efficacy of the antihypertensive regimens. PMID- 14755977 TI - [Analysis of some clinical aspects of degenerative valvular heart diseases in medical practice]. AB - In a period of 5 years there were 18,391 admissions; out of them 1129 cases were diagnosed with valvular lesions: 223 (19.7%) were degenerative valvular heart disease, 608 (53.8%) had rheumatismal valvular lesions, 7 (0.6%) had congenital valvular lesions and 291 cases (25.7%) had valvular lesions of other etiologies. Out of the 223 cases with degenerative valvular lesions, 99 cases (44.4%) were men with an average age of 70.1 years old and 129 were women (55%) with an average age of 74.9 years old. The calcific aortic valve stenosis was encountered in 139 patients (62.3%), the aortic insufficiency was diagnosed in 19 patients (8.5%), the mitral insufficiency 49 patients (21.9%) and the mitral stenosis in 10 patients (4.4%) the other patients having either aortic or mitral valvular disease. The combination of an aortic stenosis with a mitral insufficiency was diagnosed in 46 cases (20.6%) from the 223. Only 14 patients were asymptomatic, most of them having heart failure (namely, 178 patients i.e. 78%) with or without angine pectoris or effort vertigo, or they had only effort angina, vertigo or effort sincope. Rhythm disorders happened in 59 patients (26.4%) while disorders in the transmission of the stimuli were diagnosed in 14 patients (5.2%). Two patients died due to cardiac causes. CONCLUSION: Rheumatismal valve disease are nearly 2.5 times more frequent than degenerative valve disease and they became a practical reality, which is claimed by its continuously increasing frequency, by a variety of lesional aspects and by implications on the heart, and by it, presence in an age group were arteriosclerosis cumulates its risk factors. PMID- 14755978 TI - [Interactions between angiotensin-(1-7)and angiotensin II in isolated rat portal vein]. AB - Our preliminary data show for the first time the interaction between angiotensin (1-7) (Ang-(1-7)) and angiotensin II (Ang II, 10 nM) in isolated rat portal vein. Very low concentrations (10 nM) of Ang-(1-7) have marked functional antagonizing effects on Ang II-induced contractions. High concentrations of Ang-(1-7) (1-10 mM) do not affect the effects of Ang II. The effects of low concentration Ang-(1 7) might be associated to the interaction with Ang-(1-7) specific receptors and the own contractile effects (approximatively 28%) at high concentrations might be assign to the interaction with angiotensin specific receptors AT1. But, the lack of effects of Ang-(1-7) high concentrations on Ang II-induced contractions hardly might be associated to the interaction with AT1 receptors. Although losartan was entirely blocking the Ang-(1-7) effects, there is in the literature a series of data showing that Ang-(1-7) specific receptors (or a subtype of Ang-(1-7) receptors) might be sensible (with possible high affinity) to losartan. Additional experiments are thus necessary to further clarify these interactions. PMID- 14755979 TI - [The effects of the physical training on the hypertensive patients with glycoregulation disorder]. AB - The purpose of our clinical study was the evaluation of moderate physical training on hypertensive patients with the impaired glycoregulation, for a determinate period of time. METHODS: The study included 80 patients with moderate hypertension, divided in two groups: group B formed of 40 patients that followed an associated treatment (physical exercises 3-5 times/week, 45-60 min. for one training and antihypertensive drug therapy) and group C formed of 40 patients that followed only drug therapy for controlling blood pressure. The glycoregulation disorder was appreciated by determining a jeune glycemia to all the patients, the oral glucose tolerance test at the patients who were until 60 years old and for which there wasn't any evidence of glucose intolerance a jeune. The uniform pattern of OGTT values was considered due to insulin resistance and hyperinsulinism. We obtained the values of cholesterol, triglycerides, HDL-C, LDL C and atherogenesis index. RESULTS: The patients with impaired glycoregulation had significant increased values of cholesterol (p = 0.03) and triglycerides (p < 0.001), decreased HDL-C (p = 0.004), compared to the patients without glycoregulation disorder. LDL-C didn't vary significantly (p = 0.2). The patients with impaired glycoregulation had the value of ventricular mass 254.46 +/- 38 g, compared to 239.52 +/- 41 g for the patients without glycoregulation disorder. In group B, we found 14 patients with impaired glycoregulation (35%) and 11 patients in group C (27.5%) with the same metabolic disorder. After 6 month, only 9 patients with glycoregulation disorder (22.5) were left in group B and also 10 patients (25%) with the same disorder in group C. CONCLUSION: Moderate physical exercise associated with antihypertensive therapy induced, besides the decrease of blood pressure values, a good control of lipids and glucose markers of metabolism. PMID- 14755980 TI - [Obesity prevalence in Iasi county]. AB - Our study is a part of a national study initiated by ARSO (Romanian Association for Study of Obesity). We selected a statistically representative sample (1500 cases). Study was built upon a questionnaire, made by primary care physicians. Anthropometrical data evaluated the obesity (weight, waist circumference and body mass index, BMI) and we considered associated risk factors (cholesterol levels, diabetes, and hypertension). BMI, more than 25 and 30, defines overweight and obesity. Waist more than 80 cm, for women and 94 cm for men, was considered abdominal obesity. Prevalence of obesity was globally 28.69%, by BMI and abdominal obesity, 57.87%. Non-obese cases are mostly on 20-29 years (2:1), both sex and 30-39 years, mostly on women. After these decades, percents for overweight and obesity are, constantly, more on women (48%) than men (31.03%). Abdominal obesity rises constantly with age, between 42.8% and 73.04%. A lot of patient didn't know theirs cholesterol values, over 25%. The highest percent of high cholesterol is located on 50-59 years (42%) and 35.63% on previous decade. High number of menopausal women can explain this. CONCLUSIONS: 1. We consider a high prevalence of obesity, 28.69%, predominant on women and aged persons. 2. Abdominal obesity has higher prevalence, 57.87%. 3. There are multiple risk factors, especially on 50-59 years decades but are not searched sufficiently. PMID- 14755981 TI - [Importance of mild dyslipidemias determining the cardio-vascular risk factors]. AB - The aim of our study was to correlate "medium" dyslipidemias with cardio-vascular diseases, especially essential hypertension (EHT). We selected 488 cases using randomization, from a population of hypertensive people. About 210 cases benefited by lipidograms and risk assessment, using special diagrams. When cholesterol was 200-250 mg/dl, 72% of cases had EHT and over 300 mg/dl, 57% had EHT. Comparing LDL chol with EHT, we obtained that arterial tension over 160 mmHg is correlated with LDL over 190 mg/dl on 12%, when LDL was less than 190 mg/dl but more than 150 mg/dl, we found EHT on 16% cases. EHT was very strong correlated with HDL < 40 mg/dl, 88%. On 5% of cases the only one dyslipidemia was marked by low HDL chol and all this cases had EHT. A particular risk was represented by the dyslipidemia with low HDL chol and high triglycerides (28% of cases). We concluded that the highest prevalence was for medium hypercholesterolemia with/or low HDL chol and, the same time, extreme values had comparable prevalences with medium values. PMID- 14755982 TI - [Electron microscopy study regarding the etiology and the pathogenesis of acne vulgaris]. AB - The etiology and the pathogenesis of acne vulgaris are not complete cleared up yet. This work presents the results of the electronomicroscopic examinations effected on lesions of acne vulgaris biopsied from 5 patients with different clinical forms of disease (including acne fulminans). In this study we had in view the investigation and description of the ultrastructural modifications that followed the intervention of the 4 factors incriminated in the etiology and pathogenesis of acne: sebaceous hypersecretion, hyperkeratosis pilosebaceous infundibulum, bacterial colonisation, perifollicular inflammation. The ultrastructural aspects that we had in view underline the role of the 4 etiology and pathogenesis factors of acne vulgaris, confirming the data related by the specialised literature. PMID- 14755983 TI - [Secondary effects of chronic treatment with levodopa in Parkinson disease]. AB - We have analysed the side effects of levodopa preparations in patients suffering from Parkinson's disease. The study has begun in 1993. All the patients were initially admitted in the Neurological Clinic of the Recovery Hospital in Iasi, and then followed in ambulatory condition. We have monitorized the apparition of adverse reactions of levodopa therapy, and tried to treat them by modifying doses and patterns of administration, by associating other drugs or by replacing levodopa with other drugs. We have initially included 116 patients, of whom 30 have developed secondary effects until June 2002. The most frequent secondary effects have been coreea, dystonia, mental adverse reactions, yo-yo-ing etc. Each one of the 30 patients has developed one or more types of secondary reaction. PMID- 14755984 TI - [Epilepsy in different types of cerebral palsy]. AB - Epilepsy (E) is common in all types of cerebral palsy (CP) and in some varieties it is the rule. The aim of this study is to evaluate the nature, course and prognosis of seizures in patients with E and CP. Clinical observations were performed on a group of 109 patients, from 2.5 to 17 years of age, 2/3 associated mental retardation. The results proved that all types of seizures can be observed at the patients with CP and the epileptic attacks can be controlled in most cases with polytherapy. PMID- 14755985 TI - [Hand reconstruction by microsurgical free toe transfer]. AB - Reconstruction of complex hand mutilations with multi-digital or thumb amputations are best treated with microsurgical toe transfers. We present the results of the first 15 cases operated by the first author, of which 12 are thumb reconstructions (6 great toe and 6 second toe transfers) and 3 long fingers reconstructions with combined second and third toe transfers. There were no microsurgical complications. Cortical integration and functional integration was achieved for all transferred toes, with discriminatory sensibility (m2PD between 5 and 13 mm) and active mobility range between 30 and 60 degrees. PMID- 14755986 TI - [Minimal invasive treatment of the adnexal surgical pathology]. AB - Laparoscopic approach has become the "golden standard" in managing a wide range of adnexial sufferings. Most laparoscopic cases allow patients to recover quickly. Conventional ultrasonography appears to be useful in the preoperative selection for this surgery. Conventional pulsed Doppler and colour Doppler sonography proved their importance in the diagnosis of malignancy. The aim of the study is to evaluate the advantages and the limits of the minimal invasive approach in managing surgical adnexial sufferings. A retrospective study was carried on 75 operative laparoscopy cases performed between 1998-2002. Surgical procedures were performed mainly for ovarian cysts (69 cases, 92%), but there were also treated other types of adnexial pathology such as: hydrosalpinx (2), piosalpinx (1), tubal cysts (2), ectopic pregnancy (7), pelvic adherences (7) and one case of hydatid tubal cyst. Types of surgery performed were cystectomy (36), ooforectomy (11), salpingo-ooforectomy (10), fenestration (10), salpingectomy (6), lysis of adhesions (7). The mean operation time ranged between 50 and 80 minutes and the mean postoperative stay was of 3.65 days. PMID- 14755988 TI - [Caliceal stones:What did we learned after 10 years of ESWL (extracorporeal shock wave lithotripsy)]. AB - Since its first clinical application in 1980, the indication of ESWL has rapidly extended from renal pelvis stones to almost all urinary stones. The authors discuss about their experience accumulated in the past ten years, when 476 patients with caliceal stones were treated by ESWL. The global stone free rate was 60.05%, but almost a third of the patients (where fragmentation was radiological documented after ESWL) did not returned back for control. ESWL is a good solution for superior and middle calix stones less than 1.5 cm, not very opaque, with a sufficient large caliceal width. For the stones in the lower calix, the most important criteria for success are: the stone size, the opacity and, for sure, the angle between infundibulopelvic and ureteropelvic axis. PMID- 14755987 TI - [Ureteral stenting after retrograde ureteroscopy. Experience of Iasi urological department]. AB - Ureteroscopy is a well-established procedure, which has proven the efficiency for diagnostic purposes, but mostly for ureteric stone removal. The authors describe their strategy after ureteroscopic lithotripsy with Wolf 8 Ch semirigid ureteroscope. Insertion of a double J stent for 2-4 weeks was the rule when there was an important bleeding during the procedure, the fragments were big and could not be extracted, some fragments migrated in upper third of the ureter or renal pelvis, a perforation occurred or it was found a ureteral stenosis. If the stone can be extracted without any problems (in one piece) and the ureter is normal there is no need for ureteral stenting. Having in mind that many patients describe problems with the double J stent (pains, polakiuria, etc) we recommend to insert a stent at the end of ureteroscopy only for selected cases. PMID- 14755989 TI - [Risk of increase in nosocomial infections incidence in a surgical clinic. A case control study]. AB - The complex activities of epidemiological surveillance of nosocomial infections (NI) have, among the main objectives, the identification of the causes and the assessment of risk factors. In 2002, the Clinic Hospital of Pneumology Iasi had 220 beds and was divided into the following units: Pneumophtisiology Unit (PPU), Thoracic Surgery Unit (TSU) and Intensive Care Unit (ICU). The global incidence of NI was of 0.85%. The incidence of cases with NI in the TSU and ICU increased to 5.39% in the interval April-June 2002 so that urgent epidemiological measures were applied to limit the phenomenon. After that, a case-control study was proposed in order to identify and assess the risk levels. The retrospective study met the methodological conditions such as case definitions for NI and post surgical NI, for extrinsic and intrinsic risk factors, the selection of cases according to inclusion and exclusion criteria, the formation of the representative sample. The study included 16 patients with pleural NI and 55 controls with the pleurotomy performed between April-June in the TCU, as a common feature. The factors that increased the probability of a post-surgical NI development were: the external pleural drainage maintained more than 30 days (OR = 185.5; p < 0.0001 for CI = 95%), tuberculosis as the most important associated disease among the patients' pathological history records (OR = 28.0; p < 0.0001; CI = 95%); the antibiotherapy with multiple associations (OR = 3.30; p < 0.04; CI = 95%). The conclusions underlined that the patients suffered from tuberculosis since the very admission, which need pleurotomy have an increased risk to develop a NI. This fact should require an appropriate epidemiological, clinical and microbiological surveillance and the empirical antibiotic strategy or that conducted by the antibiogram results has to be performed adequately, according to the operative recommendations. PMID- 14755990 TI - [Female fertility in transition period]. AB - In the last period, women fertility declined significantly. A series of factors were taken into account in a survey that included a number 440 women from Iasi county. More than half of the women have one (27.14%) or two children (30%) and almost 55% of women are satisfied with the number of children they already have. The main factors that seems to influence the fertility are: women age, duration of marriage and women profession. PMID- 14755991 TI - Hypolipidemic effect of a prodrug containing nicotinic acid in rats. Correlation with plasmatic levels. AB - We have studied a macromolecular prodrug that contains nicotinic acid bound on a polymeric support of dextran. We have determined plasmatic levels of nicotinic acid, by a HPLC method, after macromolecular conjugate administration for a 24 hours period. In order to study the hypolipidemic effect, triglyceride levels were determined and correlated with plasmatic levels of nicotinic acid. The obtained data show that, starting from 6 hours after polymeric conjugate administration, plasmatic levels of nicotinic acid are high enough to determine a significantly decrease in triglycerides level. As a conclusion we assume that the active substance was gradually released from the polymeric support leading to a prolonged presence of the active substance in the body, which caused a lowered triglycerides level. PMID- 14755992 TI - [Malondialdehyde and free thiol groups from smokers' plasma]. AB - The nicotine and oxidants from cigarette smoke, could generate oxidative stress in human body. In our study, we determined malondialdehyde and free thiol groups in smokers plasma, the experiment was effectuated to blood donors, smokers (20 persons) and non-smokers (12 persons). The determinations was effectuated with 2 thiobarbituric acid for malondialdehyde and with 5,5'-DTNB for thiol groups. The results was expressed in mmol/L for malondialdehyde and mmol/ml for free thiol groups. Smokers presented higher values for malondialdehyde (1.519 +/- 0.683), than non-smokers (1.362 +/- 0.664). The free thiol groups values was decreased at smokers (0.502 +/- 0.136), than non-smokers (0.517 +/- 0.120). Statistically, insignificant differences (p > 0.05) between smokers and non-smokers, for both parameters, could be determined by the small number of cigarettes smoked (12-15 cigaretts/day), also by the alcohol, because all persons drunk occasionally alcohol. PMID- 14755993 TI - [Pharmacognostic research of some species of Achillea. Note 1. Volatile oils analysis]. AB - We established the hydrodistilled essential oil yields from the dried flower heads of A. collina J. Becker ex Reichenb. (collected in Austria and Romanina), A. millefolium L. (s.l.) (Romania) and of three mixtures of some species belonging to the Achillea millefolium L. group. Constitutents of the essential oils were studied by GC method. The main components of the monoterpene fraction of the oils were 1.8 cineol, camphor, and borneol. The main active principle of the volatile oil, chamazulene, was in highest concentration (53%) in the essential oil hydrodistilled from A. collina collected in Austria. The results obtained for a mixture of some species (Plafar Botosani) differ from literature data in the presence of bisabolol, which have not been found so far in Achillea species. PMID- 14755994 TI - [Research concerning rutin semisynthetic derivatives, synthesis and pharmaco toxicological properties of some new morpholine and piperidine derivatives]. AB - In order to continue the research of the rutin semisynthetic derivatives we obtained some new morpholine and piperidine derivatives. Melting points, solubility, yield, C, H, N elemental analysis and UV spectra characterised all of these compounds. The toxicity was established by intraperitoneal administration at mice; the values of LD50 are between 743.75 and 856.25 mg/kg (medium toxicity). Pharmacological assays included the following parameters: leukocytes formula, NBT test, and serum complement activity. The new derivatives of rutin have a very good solubility in water, a pH closed to physiological value and immunosuppressive activity: peripheric blood PMN and lymphocytes are decreased; fagocytic capacity of peripheric blood PMN is also decreased; serum complement activity is not changed. PMID- 14755995 TI - [Upper gastrointestinal bleeding from a perforated gastric ulcer into left atrium, after esophageal resection and gastric replacement of the esophagus]. AB - Using the stomach as a substitution after oesophagectomy is the most common method. The stomach brought intra thoracic it seems that maintains or regains its capacity to secrete hydrochloric acid and therefore can develop specific conditions, despite total denervation following bilateral troncular vagotomy. We are presenting the case of a young patient who was operated on for a corrosive esophagitis. She had an oesophagectomy and a transposition of the stomach to the posterior mediastinum and anastomosed to the cervical esophagus. She presents with upper gastro-intestinal bleeding from gastric ulcer penetrating into the left atrium. PMID- 14755996 TI - [Small size cephalo-pancreatic cancer. Demonstration of MRI/MRCP usefulness]. AB - 48 years old woman with obstructive jaundice has been examined by abdominal ultrasound followed by MRI. The suspicion of cephalic pancreatic cancer raised by ultrasound was confirmed by MRI and MRCP. Association of MRCP with axial and coronal T1 and T2 sequences successfully demonstrated a resectable tumor, confirmed at surgery. MRI/MRCP is demonstrating great potential in predicting resectability of cephalic pancreatic tumors. PMID- 14755997 TI - [HELLP syndrome--a severe form of pregnancy-induced hypertension. A case report]. AB - HELLP syndrome consists in serious liver alterations during pregnancy induced hypertension, that causes hemolysis, elevated liver enzymes and low platelet count. It is frequently accompanied by evidence of severe kidney and brain involvement. Our paper aims to stress the importance of an early accurate diagnosis and an adequate management by presenting an illustrative case. PMID- 14755998 TI - [Hypertrophic cardiomyopathy--cause of sudden death]. AB - Hypertrophic cardiomyopathy (CMH) is defined by the absence of left ventricular dilatation and the presence of myocardial hypertrophy that is not due to another recognised cause of hypertrophy such as systemic hypertension or aortic stenosis. The bizarre histological appearance of the myocardium is the hallmark of CMH. Myocytes and bundles of myocytes are malaligned and fibrosis may be extensive. Disarray is maximal in areas of macroscopic wall thickening. The condition is most easily recognized in a series of transverse, short axis slices across both ventricles. Histological examination of sections taken in this transverse plane at all three levels (high, mid, apical) including septum, anterior, posterior and lateral walls are needed. The authors report a case of cardiac sudden death to a 45 years old white man. The diagnosis was revealed intra vitam and it was confirmed post mortem. PMID- 14755999 TI - [A report of generalized secondary peritonitis]. AB - Acute peritonitis not treated within the first 24-48 hours leads to complications that have to be solved by the surgeon. These complications represent MOF and the prognosis of the case is reserved. Our case has the following features: young female of 29 years of age admitted into the hospital in the gynaecologic department as she was in pain in the hypogastric area with suspicion of diagnosis by bilateral metroanexite. She had her coil devise removed three weeks before. After wards she followed the prescribed treatment with powerful antibiotics. After six days of medical treatment, she suffers of abdominal meteorisma and vomits, she laks fever and has leucocytes 6200/mmc. The surgeon performs the physical examination and confirms the diagnosis of occlusion; X-ray show high hydroaeric level. The patient is operated after a short preparation. We found inside the abdomen a gangrenous appendix, a lot of pus in Douglas space and right tubar salpinx very inflamed. We performed the apendicectomy, the right salpingectomy and lavage and multiple drains. The patient left the surgical department two weeks after and in good shape. Our conclusion is that the case had a wrong diagnosis, it was misdiagnosed as the patient had dynamic occlusion. The clinical/physical examination was the decisive factor in taking the decision to operate. PMID- 14756000 TI - [Visible spectrophotometric assay of ranitidine]. AB - Ranitidine, belonging to H2-antagonist group, is a compound containing a furanic moiety and is used in peptic ulcer therapy. This paper debates the possibility of developing a new visible spectrophotometric assessment by using the reaction between ranitidine and eosine. We carried out our determinations at 505 nm, where the absorbency of ranitidine-eosine complex is maximal, and we have established the optimal reaction conditions. This method was successfully applied for ranitidine assay from pharmaceutical dosage forms. PMID- 14756001 TI - [Membrane-selective electrode for atropine assay]. AB - One membrane-selective electrode with PVC matrix for atropine is described, with atropine tetraphenylborate as electroactive material. This electrode has a linear response in the concentration range of 10(-2)-10(-5) M atropine sulphate, with a detection limit of 3.29 x 10(-6) M. This electrode was used with good results for quantitative assay by direct potentiometry and potentiometric titration of atropine sulphate from pharmaceutical formulations. PMID- 14756002 TI - [Schiff base type onno spectrophotometric determination of magnesium]. AB - By condensing hydrazide salicylic acid with o-hydroxy carbonilic compounds it was obtained 2--hydroxy acetophenon--salicyl hidrazina, Schiff base--type ONNO. These Schiff bis bases present a good capacity of complexing the Mg (II) ions. The above mentioned Schiff base forms complex with Mg (II) cation, with maximum absorbance at 386 nm, and molar absorbtivity (epsilon) = 6.27 x 10(4) mol-1.L.cm 1. The complex with Mg (II) presents a maximum stability at pH = 9.6 - 10.0. The calculated apparent stability constant is K = 3.18 x 10(3). The absorbance is proportional to Mg (II) concentration in the range of 10-60 mg/mL. In this range, the Lambert--Beer law is respected, the linearity coefficient being 0.9818, SD = 0.83, RSD = 0.84 (n = 6). The results obtained for spectrophotometric determination of Mg (II) using this Schiff base as reagent were successfully applied to pharmaceutical products containing Mg (II) cation. PMID- 14756004 TI - [Modern epidemiology facing infectious and non-infectious diseases]. PMID- 14756003 TI - [The history of experimental diabetes in Iasi ]. AB - Department of Pathophysiology, University of Medicine and Pharmacy, Iasi, Romania has a long tradition in studying the diabetes mellitus. The series of experimental studies in diabetes was open by Iulius Nitulescu. His work was focused on the carbohydrate metabolism, and the paper entitled "Sur le metabolisme du sucre chez les vieilliards. L'epreuve de l'hyperglycemie alimentaire" was published in 1933. Later, in 1965, Aurel Sneer introduced the experimental model of diabetes mellitus induced by the alloxan. In 1970, it was for the first time in the medical literature when the importance of lysosomal enzyme discharge was revealed in alloxan-induced diabetes. In the early 70s, the factors conditioning the carbohydrate homeostasis were studied. Starting with 1990, the research was directed to investigate the role of microelements, zinc and magnesium, in the pathogenesis of beta cells injury and chronic complications of diabetes mellitus. Since 1993, our studies have been focused on the role of oxidative stress in the pathogenesis of chronic complications of diabetes mellitus. The effects of exogenous antioxidants, probucol and vitamin E, on the carbohydrate and lipid metabolism, on the markers of oxidative stress, and on the platelet activity were investigated in alloxan-induced diabetes. More than 50 scientifical papers on experimental diabetes and four monographs were published in the last 40 years by the researchers of our department. PMID- 14756005 TI - Therapy of chronic hepatitis B: today and tomorrow. AB - Hepatitis B virus infection represents an important health problem all around the world, with almost 350 million people chronically infected and 1 million death per year. Despite an efficient vaccination program and despite therapeutic progresses, chronic hepatitis B is still a disease with severe evolution and an increased prevalence in many countries, including Romania. Today, we have two groups of drug that are used for chronic hepatitis B treatment: interferon and lamivudine. Unfortunately, long term results for interferon or lamivudine treatment are far from satisfactory, fact that explains the interest for new antiviral agents, which are under evaluation procedures in many clinical trials. A combination of two or more new antiviral action agents it is believed to be the optimum therapy of chronic hepatitis B in the future. PMID- 14756006 TI - [New data about neuroimmunomodulation and its functional implications]. AB - After a brief presentation of the immune system as sensorial and effector organ, which recognizes and defends against cellular aggressions, the main psychoneuroendocrine components of immune reaction regulation and modulation are shown. Both central nervous structures that control the hormonal emissions, the vegetative innervation of the lymphoid organs as wells as the afferent neurohumoral pathways involved in the making of the self-regulating and neuromodulating circuits of the humoral and cellular immune responses are mentioned. An important position is held by the interrelations between the hypothalamus-pituitary-corticoadrenal gland, the sympathetic-parasympathetic efferent pathways and the chemical messengers (hormones, neurotransmitters, interleukins, neurotrophins) which make possible the bi-directional neuroimmune communication for maintaining the homeostatic balances on this third effector pathway, too. Also experimental proof concerning the ability of central neurons to secrete neuromodulator cytokines and the presence of specific receptors for the various neuroactive molecules within lymphoid organs and circulating lymphocytes are presented. To close, the psychoemotional components of the neuroimmunomodulator circuits are mentioned, using as examples the changes induced by stress generally and oxidative stress in particular. PMID- 14756007 TI - [Alternative pathways for the activation of the renin-angiotensin system]. AB - The knowledge of the structure and function of the components of the renin angiotensin system (RAS) is rapidly increasing. The classic pathway of angiotensin (Ang) II generation includes a reaction catalyzed by angiotensin converting enzyme (ACE). We actually discuss the alternative pathways for the generation of Ang-II and other angiotensin peptides (Ang III, Ang IV, Ang (1-9), Ang (1-7), des-Asp-Ang I) and the responsible enzymes (tonins, cathepsins, chymases, aminopeptidases, dipeptidyl aminopeptidases, neutral endopeptidases, carboxypeptidases, ACE2 etc.). The development of novel enzyme inhibitors (e.g., nafamostat, sampatrilat) for more efficacious suppression of RAS activity is also considered. PMID- 14756008 TI - [Severe asthma: clinical and pathogenic aspects]. AB - Difficult asthma is an uncommon disease (likely < 5% of total asthma). It is poorly understood and often difficult to treat. The first step is to make an accurate differential diagnosis and to avoid the most important factor, which influence the condition of the disease. Between these factors, the exposure to allergens and the gastro-esophageal reflux seems to be the most important. The term is usually applied to conditions that are hard to solve, sometimes dangerous or even life-threatening forms of asthma: corticoid-resistant asthma, corticoid dependent asthma, brittle asthma, premenstrual asthma and near-fatal asthma. PMID- 14756009 TI - [Implication of methylglyoxal in diabetes mellitus]. AB - Methylglyoxal (MG) is an early glycation product which is implicated in genesis of diabetic complications either as a direct toxin or as a precursor for advanced glycation end products. It is metabolized via S-D-lactoylglutathione to D-lactate by means of the enzymes glyoxalase I and II, which depend on glutattione as cofactor. MG is highly reactive and can bind to and modify proteins by chemical interaction with cellular proteins, action on energy production, induce free radical generation and cell killing. That way MG play a role in the events of the development of diabetic complications. PMID- 14756010 TI - [Treatment of cholestatic liver disease]. AB - Primary biliary cirrhosis (PBC) and primary sclerosing cholangitis (PSC) are chronic cholestatic liver diseases, biochemically characterized by an elevated serum bilirubin, alkaline phosphatase and gamma-glutamyl transferase. Although PBC and PSC have their own distinctive clinical, immunological, radiological and histological features, both diseases show a necro-inflammatory process, predominantly confined to the bile ducts within the portal tracts. In most patients the disease gradually progress and may lead to cirrhosis and liver failure. Transplantation may than be the only therapeutic option left. PMID- 14756011 TI - [Drug residues in food of animal origin]. AB - This paper reviews literature data on the stability of drug residues in food of animal origin. It was investigated the most important antibacterial, antihelminthic and sedative drugs and growth promoters used in food/producing animal treatments. The effects of storage (at -20 degrees C, 4 degrees C and temperature room) and a range of cooking processes (boiling, frying, roasting, grilling, microwaving) on residues was studied. The results are summarized in tables and showed that degradation and biological activity were varied between 0 and 100%. PMID- 14756012 TI - [Granular cells of the hepatic capillary sinuses--receptors]. AB - Pit cells or hepatic natural killer (NK) cells represent an organ-associated NK cell population. They are situated in liver sinusoids and exert high spontaneous cytotoxic activity against tumor cell lines and may act as a primary defense barrier to metastasing tumor cells and to virus infections. Pit cells express two types of receptors on their cell membrane. One type activates NK cell killing (NCR or natural cytotoxicity receptors, as NKp46, NKp44, NKp30, NKG2D) by recognizing ill-defined molecules on target cells. The second type of receptors inhibits the lytic pathway by recognition of self class I MHC molecules and are represented by KIR or killer cell Ig-like receptors, as KIR2D, CD94/NKG2. Pit cells express on their cell membrane and other type of molecules as CD2, CD54, CD11a/CD18 and are CD3 negative. PMID- 14756013 TI - [Super-oxide dismutase and catalase in smokers]. AB - In this work, the results of a study concerning the variation of superoxid dismutase (SOD) and catalase activities to smokers are presented. The study has been performed on volunteers, blood donors, men, between 20-60 years. In the smokers group, 20 donors who smoke over 10 cigarettes/day over 5 years have been included. In the control group, 12 donors who never smoked or stopped to smoke over 4 years have been included. SOD and catalase activities have been determined spectrophotometrically from erythrocytes lysis; the enzymatic activity has been reported to g Hb. The results show a decreasing of SOD activity to smokers reporting to the control group, totally with 17.7% (from 69.6 +/- 18.6 to 57.3 +/ 26.6 U/gHb). Comparing with control group, the catalase has been induced to smokers with 0.04%, from 8.82 +/- 3.32 to 9.18 +/- 8.66 x 10(3) U/gHb. Statistically, the variations are insignificant and we suspect the interferation of alcohol as an important oxidative stress inductor. PMID- 14756014 TI - [Systemic treatment of colorectal cancers--factual standards and perspectives]. AB - Adjuvant therapy has been shown to reduce recurrence and improve survival in patients with stage III colo-rectal cancer (CRC). However, the use of adjuvant therapy is still much debated in stage II disease. Fluorouracil (5-FU) and folinic acid (FA) are currently the standard adjuvant drug combination. The treatment of patients with metastatic colorectal cance has changed dramatically over the years. The more optimal use of 5-FU in association with FA, the new drugs such as irinotecan and oxaliplatin, and the oral fluoropyrimidines capecitabine and uracil/tegafur(UFT) have contributed to the increased therapeutic option and to improved outcome of patients with metastatic CRC. It has been shown that combination therapy with 5-FU/FA and irinotecan or oxaliplatin is more active than 5-FU/FA in the first line of advanced CRC. The oral fluoropyrimidines capecitabine and UFT/FA seem to have a comparable activity to intravenous bolus 5-FU/FA in the first line treatment of metastatic CRC. New agents acting on novel targets are under development. Epidermal growth factors inhibitors, vascular endothelial growth inhibitors, and cyclo-oxygenase 2 inhibitors might play a role in the future in the treatment of CRC. PMID- 14756015 TI - [Epidemiologic and clinical aspects of acute hepatitis B in the past decade]. AB - To study the clinical and epidemiological aspects of acute B hepatitis during the last 11 years. We retrospectively studied 1712 patient files, admitted in the Department of Infectious Diseases Iasi, with acute B hepatitis between 1992-2002. The majority of the patients (69%) had an urban origin. Teenagers and young adults were predominantly affected (59% had between 15 and 34 years). A point of entry of the pathogen was identified only in 20% of the patients. The mean incubation period was 4 month. 24% of the patients had a prolonged form of the disease (over 30 days of jaundice). A fulminant evolution was noted in 1.2% of cases. Extrahepatic involvement was described in 25% of the patients. The global mortality was 1.15%. The number of patients with acute B hepatitis decreased by half during the last 11 years. PMID- 14756016 TI - [Acute viral encephalitis. Experience of the Iasi Infectious Disease Clinic between January 2000 and March 2003]. AB - We analyzed the patients files suffered from acute viral encephalitis in 39 months period: January 2000-march 2003. According to our data there was a total of 33 patients; one of them died. The most involved patients were male and of the 8-14 year group of age; for the first three month in 2003 were admitted 11 patients. Coma and seizures were the most frequent clinical signs. LCR had the usual abnormalities for the above mentioned disease; the treatment was mainly performed with corticosteroids; 28 cases were admitted more than 8 days period. The evolution of the clinical status was satisfactory for all 32 patients. PMID- 14756017 TI - [Is physical training useful for elderly patients with ischemic heart disease?]. AB - The aim of the paper is the study of the effects of a supervised physical training at old patients with ischaemic heart disease. METHODS: 74 patients with ischaemic left ventricular dysfunction have been followed during a supervised physical training consisting of a daily programme of exercise over a period of 21 +/- 10 days. They were parted in Group A (n = 26) over 60 years old and Group B (n = 48) under this age. The heart rate (HR), systolic and diastolic heart pressure (SHP respectively DHP), the time-tension index (TTI) were registered during the rest, the peak intensity of exercise and the recovery. These parameters were compared in evolution at each group and also between the groups. RESULTS: At Group A, at the end of the period, SHP at the peak intensity decreased (from 146 +/- 19 to 137 +/- 14 mmHg, p = 0.01), also SHP in the recovery period (from 119 +/- 10 to 116 +/- 10 mmHg, p = 0.01) and HR during the recovery time (from 77 +/- 8/min to 73 +/- 6/min, p = 0.03). The total length of the session time increased (from 12 +/- 4 min to 21 +/- 6 min, p = 0.01) and also the duration of the peak exercise time (from 3.6 +/- 1.0 min to 7.7 +/- 2.9 min, p = 0.03), in the condition of maintaining a HR at the peak exercise of about 75 76% of the maximum HR obtained at a previous exercise testing. The improvement of the training parameters was similar with Group B, except the duration of the session, longer at the last group (21 +/- 6 min at Group A vs. 25 +/- 7 min at Group B, p = 0.02) and the duration of the peak intensity (7.7 +/- 1.0 min at Group A vs. 9.1 +/- 2.9 min at Group B, p = 0.02). CONCLUSIONS: Old patients with ischaemic left ventricular dysfunction improve their exercise capacity as well as the younger ones; the benefits of a supervised physical exercise are evident regardless of age. PMID- 14756018 TI - [Fever of unknown origin in Romania. II. Diagnostic Procedures. Prospective multicenter study of 164 patients]. AB - BACKGROUND: The Diagnostic workup of patients with fever of unknown origin is a challenge, due to the great number of possible etiologies. After we studied the etiologic spectrum of fever of unknown origin in Romania, we tried to evaluate the diagnostic procedures used and their efficiency. METHODS: A multicenter cohort study of two years, with another two years of follow-up was carried out on 164 consecutive patients who met the classic, modified criteria of fever of unknown origin. We used a standardised diagnostic protocol. MAIN OUTCOME MEASURED: The role of every diagnostic procedure in establishing the final diagnosis. RESULTS: The diagnosis was made by microbiology and serology in 41 cases (25%), by histopathology in 22 cases (18%), with the help of imaging techniques in 30 cases (1.3%), based on the clinical evolution and response to treatment in 54 cases (33%) and by other methods in 12 cases (7.3%). The abdomino pelvic ultrasonography had a sensitivity of 60%, a specificity of 70%, a positive likelihood ratio of 2.02 and a negative likelihood ratio of 0.57, while the scanner had a sensitivity of 81%, a specificity of 64%, a positive likelihood ratio of 2.23 and a negative likelihood ratio of 0.29. CONCLUSIONS: Of all the diagnostic procedures used, none had a good sensitivity/specificity. The clinical evolution and the treatment response had an important role in the diagnostic workup. PMID- 14756019 TI - [Direct detection by ligase chain reaction of Mycobacterium tuberculosis complex in pleural fluid]. AB - Our purpose was to investigate the utility of the DNA amplification by ligase chain reaction (LCR) for the direct detection of Mycobacterium tuberculosis complex (MTC) in pleural fluid specimens from patients suspected for tuberculous pleural effusion. We have used the LCx M. tuberculosis kit (Abbott) which uses the amplification of the gene that encodes for antigen b. We have examined 81 pleural fluid specimens by isolation (on Lowenstein-Jensen medium and MB/BacT system) and by LCR. Out of 10 positive specimens in culture, 4 were also positive by LCR; out of 71 negative specimens in culture, 8 were positive by LCR. We have re-evaluated the LCR results according to the clinical diagnosis, sustained by the successful therapy, and to the pathological diagnosis on the pleural biopsy. The sensitivity and specificity of LCR in the diagnosis of tuberculous pleural effusion were 31.5% and 100%. This commercial LCR kit is a rapid, specific, but less sensitive test for the routine diagnosis of the tuberculous pleural effusion. PMID- 14756020 TI - [Global survival rate calculated by Kaplan-Mayer method in children with malignant lymphomas]. AB - The Kaplan Maier method is being used in oncology in order to calculate the survival rate during/at the end of the study. The purpose of this study is the assessment of the survival period referred to the clinical and histopathological state, laboratory findings, the diagnostic and treatment time. The material of the study is a LIMCO group (n = 308 cases) hospitalized in the Oncology Department of the Children's Clinical and Emergency Hospital "Sf. Maria", Iasi between 1980-1995. The LIMCO group was divided in 2 smaller groups: LMH group (110 cases) and a LMNH group (198 cases). The results statistical analysis was made by chi 2, Long-Rank and Breslow test. The results established a series of favorable LIMCO prognostic factors referred to the clinical state I-II, the histopathological type, normal or pathological laboratory findings at the first admission and the quality of the remission after the multimodal treatment. PMID- 14756021 TI - Statistical epidemiological screening methods assessing certain biochemical markers. AB - The epidemiological studies have shown that it is difficult to define a "normal" value for the serum concentration of some biochemical markers with predictive significance for cardiovascular diseases (CVDs) as follows: total cholesterol (TC), total lipids (TL), triglycerides (TG), glycemia (G), and urea (U). Our study establishes the mean and limit values of the normality for the above mentioned markers, in an adult population from the Iasi City, using the epidemiological screening method. The employed methodology includes the records of the results of 2788 biochemical determinations during a period of 5 years, in order to assess the state of health of the population, as well the data working up and interpretation by means of the EpiInfo computer program. By testing a series of "normal" limits, the values found with apparent healthy persons (n = 2691) and persons previously diagnosed with diabetes mellitus or CVDs (n = 97) were analyzed in a differentiated way. In the sample of apparent healthy individuals, with the age between 20 and up to 60, the mean values were: 210 mg% for TC; 6.74 g@1000--TL; 125.74 mg%--TG; 0.91 g%--G and 0.34 g%--U. Satisfactory correlations (correlation coefficient-"r") of age with the values of TC (r = 0.38); TG (r = 0.26) and G (r = 0.23) comparatively with U (r = 0.15) or TL (r = 0.14) were noticed. The comparison of the mean values between males and females did not reveal a significant difference (p > 0.05) for any of the assessed constants. The clinical and community studies of the CVD risk assessed in the Iasi City area will have to be referred to the registered and analyzed values of this epidemiological screening in the interpretation of some biochemical markers data. PMID- 14756022 TI - [Hormonal influence on gastrointestinal reflux during pregnancy]. AB - Gastroesophageal reflux (GER) occurs in 30-50% of all pregnancies. The progressive rise in plasma progesterone has been suggested as a possible mediator of GER during pregnancy. It is not known whether progesterone, at physiological concentrations, has an effect on acid contact time. The study includes 70 women- 50 pregnant and 20 non pregnant healthy menstruating female volunteers. We used immunoenzymatic tests for determination of estradiol, progesterone and beta HCG hormones during pregnancy. The 50 pregnant women with GER symptoms heartburn, acid regurgitations had increased levels of steroid hormones. 24-h ambulatory esophageal pH monitoring and serum progesterone levels were determined in 20 healthy women known to have normal menstrual cycles. All tests were performed during the follicular phase days 2-7 and the luteal phase days 22-28 of one or two consecutive menstrual cycles. The fluctuations in progesterone levels across the normal menstrual cycle have no significant impact on 24-h ambulatory pH parameters. CONCLUSIONS: Progesterone, at physiological concentrations, does not predispose to GER in healthy menstruating women. Thus, it is likely that if progesterone, with or without estrogen, contributes to GER in pregnancy, this effect takes place only at the high concentrations observed during this condition. After delivery, GER disappear with decreasing hormonal levels and uterine volume. PMID- 14756023 TI - [Cardio-thoracic index: marker of the clinical course of alcoholic dilated cardiomyopathy]. AB - Alcoholic cardiomyopathy (ACM), a principal form of secondary dilated cardiomyopathy, can ensue from heavy consumption of alcohol over a long period of time. In harmful consumption, alcohol and its metabolites has a toxic effect on heart muscle cells. The clinical features include dilatation of the left ventricle, poor myocardial contractility and symptoms of heart failure. The heart and lung X-ray examination is required in all disease stages. The information gathered from this cheap and noninvasive investigation method, are very important in the diagnosis algorithm. In the ACM stages beginning, before the installation of the heart failure symptoms, it is possible to found normal dimensions of the heart, which is compatible with the alcoholic cardiomyopathy diagnosis. Specific for dilated alcoholic cardiomyopathy is the reversible character of cardiomegaly, objectified through the reduction of the cardio-thoracic index in conditions of alcohol abstinence and adequate treatment of the heart failure. PMID- 14756024 TI - [Role of calcium and magnesium ions in cerebrospinal fluid in alcoholic-traumatic coma]. AB - The goal of investigation was to determine the role of calcium and magnesium ions in the cerebrospinal fluid in ethylic-traumatic coma. We measured the level of calcium in the cerebrospinal fluid within simple photometric test and the magnesium level within xylidyl blue photometric test. We found a high mortality in patients with high level of calcium in cerebrospinal fluid and low level of magnesium in cerebrospinal fluid. At patients with ethylic-traumatic coma high levels of calcium in cerebrospinal fluid are caused by the excitatory amino acids cascade and increased of hematoencephalic barrier permeability. Decreased levels of magnesium in cerebrospinal fluid are associated with convulsions and a poor prognosis of the patients. These analyses are very important for establishment of prognosis in patients with ethylic-traumatic coma. PMID- 14756025 TI - [Value of steroid receptors assessment in atypical endometrial hyperplasia]. AB - To evaluate if the immunohistochemical assessment of steroid receptors is helpful in distinguishing Atypical Hyperplasia from Well Differentiated Endometrial Carcinoma, the morphological and immunohistochemical features for steroid receptors were studied in 46 cases of atypical hyperplasia alone or associated with other degrees of hyperplasia or adenocarcinoma. The estrogen and progesterone receptors were determined by the avidin-biotin complex technique using monoclonal antibodies on paraffin embedded tissue. The diagnosis of atypical hyperplasia was based on the observation of epithelial atypias and the absence of the stromal invasion. The positive reaction for steroid receptors was moderate, very variable from case to case, heterogeneous and quite similar to that observed in carcinoma. The morphological identification of the stromal invasion is the most reliable criterion in differentiating endometrial carcinoma from atypical hyperplasia. The steroid receptors assessment is not valuable in this purpose but may be useful in cases in which a hormonal therapy is recommended. PMID- 14756026 TI - [Congenital cystic disease of the biliary system in adults]. AB - Congenital cystic disease of the biliary system is a complex syndrome of ectasies of the intra-, extra- or both situation of biliary tree. This disease has an unsure etiopathogeny. It is uncommon through the third age, with a greater incidence in child, teen-ager and young adult. The goal of our study is to evaluate the symptoms, diagnosis, treatment and histological aspects of the congenital biliary cysts. We performed a retrospective study from March 1988 to July 2003 about 11 patients with this disease treated in our surgical clinic. Clinical features, methods of diagnosis and surgical treatment were assessed. All patients were females with mean age 51 years (extreme 26-77 years). The symptoms were: right upper quadrant pain--11 cases (100%), jaundice--6 cases (54.5%), fever--3 cases (27%), palpable abdominal mass--2 cases (18%), weight loss--1 case (9%). The imaging diagnosis was helpful (ultrasonography, CT, ERCP, percutaneous cholangiography and preoperative cholangiography). In concordance with Todani classification the patients were included in the following types: Ia--3 cases (27%), I b--1 case (9%), I c--3 cases (27%), IV a--2 cases (18%), IV b--1 case (9%), V--1 case (9%). All patients were operated on: after cholecystectomy and transcystic cholangiography (11 cases--100%) we performed the total excision of the cyst--9 cases (82%) with choledochal jejunostomy (Roux-en-Y)--6 cases (54.5%), choledochal jejunostomy (omega)--1 case (9%) and choledochal duodenostomy--2 cases (18%). In one case of neoplasic cyst with portal invasion we performed a cyst-jejunostomy (omega) and in one case of Caroli disease with total obstruction of the distal choledoc, the solution of choice was choledocal duodenostomy. The microscopic pathology of the cyst wall showed: chronic intramural inflammation--9 cases (82%) and the absence of the nervous intramural terminations--1 case (9%). The additional lesions was: hepatic cirrhosis--1 case (9%) and hepatic fibrosis--3 cases (27%). We found three cases with neoplasia: malign cyst with advanced local invasion--1 case (9%), pancreatic carcinoma--1 case (9%) and gallbladder carcinoma (microscopic finding)--1 case (9%). The postoperative morbidity includes biliary fistula--2 cases (18%) and wound infection--2 cases (18%). Long-term follow-up revealed cholangitis in one case- 9%. The cystic dilatations of the common bile duct is an exclusive indication for surgery as soon as it was discovered. PMID- 14756027 TI - Endovascular treatment of the traumatic rupture of the aortic isthmus. AB - The traumatic rupture of the aortic isthmus is a dehiscence of all or part of the aortic wall, occurring as a result of a closed thoracic trauma. Standard surgical technique requires left thoracotomy, aortic cross-clamping and use of the cardiopulmonary by-pass to prevent ischemic complications, in special the neurologic and visceral ones. The endovascular treatment of these lesions is a more simple solution and there are recent reports on the stent-grafting of the isthmic rupture of the thoracic aorta. We report three cases of successful endovascular repair of the aortic isthmic rupture and we discuss the emergency indications, the advantages and the limits of this technique. PMID- 14756028 TI - [Characteristics of anastomoses between the celiac trunk and the superior mesenteric artery]. AB - Duodenum and pancreas are two deep abdominal viscerae with multiple arterial sources and complex vascular relations, that impose peculiar surgical techniques. The dissection of 120 corpses and the examination of selective angiographies revealed the variants of the anastomoses between the superior pancreaticoduodenal arteries (that receive blood from the celiac trunk) and the inferior pancreaticoduodenal arteries (belonging to the collateral subsystem of the superior mesenteric artery). The pancreaticoduodenal arches are the main anastomosis around the pancreatic head, and supply the duodeno-pancreatic complex. During this study I found the absence of this anastomosis of the antero inferior duodenopancreatic arch 5 cases (4.16%), and the lack of anastomosis of the postero-superior duodenopancreatic arch in 11 cases (9.16%). In these situations, the superior and the inferior duodenopancreatic arteries give off the vasa recta and the pancreatic branches, as normally. I found different variants of accessory duodenopancreatic arches, and I analyzed their impact on the surgical techniques on duodenum and pancreas. This study pinpoints the importance of the anastomotic subsystem in the arterial supply of the duodeno-pancreatic complex, reveals the necessity of the preoperative angiographic exam in order to choose the most opportune surgical technique. PMID- 14756029 TI - [The importance of Dunn's procedure in the treatment of slipped femoral epiphysis]. AB - The slipped upper femoral epiphysis (SUFE) is a condition also known as 'coxa vara' of the adolescent, in which lesions of the growth plate are responsible for the development of sudden or gradual displacement of the femoral head. The disease is most frequent in the 10-15 years' age group, usually in boys who also have endocrine disorders. Both acute and chronic presentations are described. The clinical symptoms and especially the X-Ray features are diagnostic. Treatment options include orthopedic (conservative) measures or surgery, depending on several conditions, such as: the clinical picture, the degree of the femoral displacement, and the age of the patient. The chronic, long-lasting cases are generally associated with poor results. The paper presents our experience in using the Dunn's procedure in 7 chronic cases of slipped upper femoral epiphysis in children. It appears that surgery by using Dunn's procedure is an effective treatment in patients with SUFE. PMID- 14756030 TI - [Limitations in the histopathologic diagnosis and prognosis of neuroblastoma]. AB - Neuroblastoma is the most common nonhematopoietic solid tumor of childhood and has been intensively studied for at least 4 decades. Despite this, few predictive histopathologic clues to its behavior exist. Ages, anatomic sites of occurrence, histopathology of the tumor and clinical stage have traditionally been the only reliable prognostic factors in this disease. In the present study we analyzed these prognostic factors in 72 neuroblastic tumors and our histopathological possibilities of diagnosis. PMID- 14756031 TI - [Sono-hysterosalpingography in the assessment of tubal infertility]. AB - The assessment of the fallopian tubes patency is an important moment in the investigation for infertility. Vaginal sonographic hydrotubation was assessed in the evaluation of uterine configuration and tubal patency. In addition, technique using "agitated" saline during transvaginal sonography was evaluated. Ultrasound was more effective in detecting abnormalities of both interstitial and distal parts of the tube compared with hysterosalpingography. Sonohysterosalpingography is cheaper, may be performed in the office and also the pelvic irradiation is avoided. PMID- 14756032 TI - [Internal urethrotomy, a modern method in the treatment of urethral strictures]. AB - The aim of this study was to evaluate the internal urethrotomy in the treatment of urethral strictures, by the retrospective analysis of 54 cases. The patients' age ranged between 18-92. 35 patients (64.8%) had iatrogenic strictures and 10 (18.5%) had traumatic ones. 35 patients presented with urinary infections before the procedure. Following internal urethrotomy, the medium hospital stay was 3.15 days. There were only 7 patients with postoperative fever and no death was recorded. The follow-up period was between 1-12 months with a medium of 9 months and only 2 patients required recurrent internal urethrotomy. Out of the 54 patients, 8 had recurrent urethral stricture disease and 6 of them had posttraumatic membranous urethral strictures. Optical internal urethrotomy appears to be the ideal therapeutic method of urethral strictures, because it has a low morbidity, it is safe to perform and the postoperative results are good. The hospitalization and the recovery periods are short, allowing a quick socio professional reintegration. Most of the recurrencies occur with posttraumatic strictures. PMID- 14756033 TI - [Upper digestive hemorrhage, physiopathologic and therapeutical considerations]. AB - The study is based on the analysis of 158 patients admitted in the "Fourth Clinic of Surgery" with diagnosis of "Upper Digestive Haemorrhage" (U.D.H.) between 1998 2002, emphasising the importance of the physiopathological chains induced by the ulcer bleeding; 119 cases (76%) were diagnosed with gastro-duodenal ulcer, 20 with portal hypertension (20%), 14 subjects were with gastric carcinoma (8.8%) and 5 with miscellaneous etiologies so called "rare circumstances" of U.D.H. (two patients with Idiopathic Thrombocytopenia, one case with Dieulafoy gastric ulceration and two subjects with Haemorrhagic Gastritis). All the patients were admitted in the first moment in the Intensive Care Unit where haemostasis were successfully obtained with drug therapy adapted to the physiopathological changes induced by bleeding, in the majority of cases with light bleeding (8 cases, 5.4%) or medium bleeding (139 patients, 87.4%); only in 9 subjects (6%) with severe haemorrhage surgery was indicated (posterior bulbar ulcer in which the surgical haemostasis was necessary). In 88 patients (55%) surgery was performed as an elective procedure--74 from them had haemorrhagic gastric or duodenal ulcers with a medium amount of bleeding and 14 observations for gastric carcinoma with a chronic bleeding. In most all the cases the postoperative evolution was in good terms. Two observations with severe U.D.H. due to rupture of esophageal varices occurred on hepatic cirrhosis were out of therapeutical proof (1.2% deaths from all the admitted patients with U.D.H.). PMID- 14756034 TI - [Frequency of food additives in mass-consumed food products]. AB - In the effected study we wanted to reveal the richest foods in additives and the most frequent used additives. As material we used a sample formed from 100 food products investigated in a supermarket, and the method consisted in data registration of food additives written on the labels of the original packages with statistical analysis. We could not establish the food additive concentrations in foods because we have no adequate technology. The food groups with the most additives were: meat products (8 types), soft drinks (7), hydrogenated fats and sugar products (6). The most frequent used food additives were: for meat products--support substances (63%); soft drinks--acid, flavor (100%) and preserve (88%) substances; hydrogenated fats--emulsion, flavor, color substances (100%) and for sugar products--emulsion (85%) and flavor substances (78%). In conclusion, food additives with possible noxious effects on health are frequently incorporated in food products. PMID- 14756035 TI - [Relevance of histopathologic diagnosis in amelogenesis imperfecta]. AB - The aim of the study was the determination of the correlation between the morphostructural aspects of the enamel defects and the clinical diagnostic and the nosologic kinds of these diseases. The research was made on 600 children from Iassy between 7-11 years old, who were examined in natural light. The index used for the systematization of the data was EDS. We used 21 temporary teeth and we examined these teeth on an optic microscope. The study show that there is no absolute correlation between the morphostructural aspects of the enamel defects and the clinical aspects of the lesions, and the clinical defect may not be always associate to the dysplasia from the imperfect amelogenesis. PMID- 14756036 TI - [Investigation of antimycotic action of pyridazine derivatives]. AB - This paper describes the synthesis of two tetrazolo-pyridazines and of ditetrazolophtalazine. Also, it is given the assessment of their antimycotic activity, being noticeable a selective fungicidal action. PMID- 14756038 TI - [Acute diarrhea associated with Cyclospora cayetanensis]. AB - Acute diarrhea is a major problem with high morbidity and mortality rates in developing countries, especially in children. Complex laboratory investigations are required to define the etiology because of the broad spectrum of etiological agents and the non-specific clinical signs. In the last decade, Cyclospora cayetanensis--a new acid-fast coccidian species--was pointed to be the cause of watery self-limited or prolonged diarrhea in immunocompetent and immunocompromised patients, with very good evolution after treatment with co trimoxazole. Unlike Cryptosporidium parvum, nonsporulated Cyclospora oocysts are eliminated in feces, with no risk of human to human transmission. Cyclospora cayetanensis is widely spread, producing endemic infections, in Asia and South America and was reported to produce infections in foreign travellers in these areas and epidemic outbreaks of foodborne diarrhea. We describe the first case of acute diarrhea associated with Cyclospora cayetanensis in an immunocompetent child admitted to Iasi "Sf. Maria" Hospital. PMID- 14756037 TI - [Study of the cardiovascular properties of some new methyl-xanthine derivatives]. AB - The effects of some new methylxanthine compounds, derivatives of theophylline, theobromine and caffeine with various radicals in 8 position on blood pressure have been studied. Derivatives of theophylline (1,3-dimethyl-xanthine) with various radicals in 8 position have reduced the blood pressure. In the same experimental conditions the theobromine (3,7-dimethyl-xanthine) derivatives haven't modified the blood pressure while the caffeine (1,3,7-trimethyl-xanthine) derivatives have increased the blood pressure values. PMID- 14756039 TI - [Morphologic and clinical correlations in medullar thymoma]. AB - The medullar thymoma is a rare and distinctive epithelial thymoma, a thymic tumor characterized histologically by a mixture of spindle epithelial cells and lymphoid cells. We are presenting this tumor to a 68 years old man, admitted at CCI, for a mediastinal tumor, treated by tumorectomy, for revealing the cytological, histological and immunohistochemical characteristic features. The surgical biopsy was prepared by using usual histological techniques and haematoxilin eosin and Van Gieson stainings. We are discussing the relation between the thymoma clinicopathological and prognostic features, resulting a clear correlation between histological type and clinical study. We also pointed the Muller-Hermelink thymoma histological subtypes and their correspondence with OMS histological types, reflecting realistically the thymoma clinical behavior. PMID- 14756040 TI - [Hemangioendothelioma in children]. AB - The hemangioendothelium is a vascular tumor rarely seen in the pediatric practice. This case underlines the difficulty that lies in establishing a differential diagnosis with a localized adenopathy when this vascular tumor is developing inside a ganglionic region. The surgical treatment followed by interferon therapy determined a favorable evolution in this case because the hemangioendothelium is a vascular borderline tumor. Still, there are cases that recur, metastasize and have an evolution towards death. PMID- 14756041 TI - [Pelvic-perineal prolapse of 3rd grade after subtotal hysterectomy]. AB - The pelvi-perineal prolapse (PPP), which is a relatively well defined anatomo clinical and surgical entity, involves a complex etiology and pathogeny. The relatively rapid appearance of PPP after subtotal hysterectomy without the suspension of the vaginal stump and perineal reconstruction has already been substantiated. This study refers to a similar case, where it was necessary to apply some non-typical surgical technics (cervical stump extirpation, colpoligamentopexy with 4 non-resorbable threads at the Cooper ligaments and perineal reconstruction), with the aim of vaginal suspension, at a menopausal patient with PPP of 3rd degree, stress urinary incontinence, ulcerating remaining cervix and cystorectocele, installed at 2 years after the inter-annexal subtotal hysterectomy. PMID- 14756042 TI - [Minimally invasive treatment of pulmonary and pleural hydatid cyst]. AB - The incidence of hydatid disease is increased in Balkan region and Eastern Europe. The global incidence in Romania is 5-6 cases of 100,000 populations. The etiological agent is Taenia echinococcus. The pulmonary localization is about 30% from all cases with hydatic disease. The only efficient treatment is represented by surgical intervention with minimal visceral and parietal destruction. We will present 4 cases with pulmonary and pleural hydatic cysts that were treated with minimally invasive techniques. New minimally invasive approaches are developed to reduce physical discomfort, to offer a better intraoperative visibility, for the esthetic aspect of incisions, shorten hospitalization and quick socio professional reintegration. The mean duration of hospitalization in these 4 cases was 8.5 days (limits between 7-12 days). The rate of complication and the necessary of analgesics were reduced. The medical treatment with Albendazole 10 mg/kg/day, 14 days in preoperative and 3 months postoperative period was followed in all patients. After discharge, imaging evaluation in the first 24 months is very important. The obtained results were excellent and we conclude that minimally invasive surgery in pulmonary hydatid disease represent a better method of surgical treatment. PMID- 14756043 TI - [Primary pleural-pulmonary non-Hodgkin lymphoma]. AB - Primary pleuro-pulmonary non-Hodgkin lymphomas are very rare tumors, possibly under diagnosed, with a very long, frequently asymptomatic evolution. The authors present the diagnostic difficulties of such pathology, in a particular case, which was diagnosed only by open biopsy and immunohistochemical analysis of tissue fragments. Chemotherapy and radiotherapy determined a favorable course of the disease. PMID- 14756044 TI - [Comparing population means using the ANOVA method]. AB - ANOVA is the abbreviation of the ANalysis Of Variance. It was developed for inferences concerning more than two populations. The tested hypothesis is the equality of the means of several normal populations. Some mathematical aspects are presented and also advantages and drawbacks of the method are marked out. Finally a practical example (implementing the ANOVA test) using the SPSS statistical software and also Microsoft Excel are shown. PMID- 14756045 TI - [Modern surgical treatment of aneurysmal bone cyst using a synthetic bone substitute (Ceraform, a calcium phosphate ceramic)]. AB - Aneurysmal bone cyst is a benign pseudotumoral dystrophy of bone, presenting expansive, destructive, nonresolutive features and often recurrence. A "stand-by" therapeutically attitude, in a survey manner, based on some spontaneous regression of certain areas of the aneurysmal cyst, due to thrombosis and fibrosis, is rarely advocated. When tumoral lesions are located in long bones and the bone length must be preserved, the bioptic curettage, followed or not by auto grafting or cortical/cancellous allografting to bridge defect, represents the treatment of choice. The inconveniences encountered in using auto- or allografts have raised a growing interest towards synthetic bone substitutes. The most used of them are phosphocalcic ceramics due to their basic properties regarding interaction between bone and the substitution material, especially macroporosity Lately, we started to use (as clinical application) a bone substitute based on synthetic biphasic macroporous ceramic (CERAFORM), covering a wide range of procedures: benign tumors and dystrophies, spinal or joint arthrodesis, periprosthetic fractures, revisions following failures of primary total hip replacements. We obtained promising results, suggesting that in a situation with limited bone defects, as in clinical case presentation, if there exist a good contact and strong mechanical fixation, CERAFORM represents a reliable option comparing to allograft. PMID- 14756046 TI - [Direct mycologic examination in superficial cutaneous mycoses]. AB - The diagnosis of superficial cutaneous mycoses is firstly performed by direct microscopic exam of the pathologic specimen. The specimen needs to be correctly harvested, in a sufficient amount, from the active zones, before the application of treatment and be well performed in order to obtain a correct diagnosis. The presence of micelian filaments or of the levuric cells in the investigated material confirms the diagnosis of mycosis, without precising the involved specia. Only by correctly progressing through successive stages of diagnosis a conclusive result may be obtained. PMID- 14756048 TI - [Pleading for primary health care]. PMID- 14756049 TI - [Basic principles of the antisense strategy]. AB - The antisens strategy is based on specific inhibition of the mutant gene expression by oligodeoxynucleotides (OGN), capable of selectively hybridizing with target DNA or RNA. Molecular mechanisms of the antisense oligonucleotide comprise: inhibition of splicing, inhibition of 5'-capping and 3' polyadenylation, activation of RN-ase H, small interfering RNA, ribozymes. The antisense strategy is applied to: rational design of potent, selective therapeutic agents, target validation and detection of pathologic gene expression in vivo. The main aim of the antisense oligonucleotide design is to improve the affinity for target RNA and the resistance to nucleolytic degradation. PMID- 14756050 TI - [Physical exercise and vascular endothelium]. AB - Physical exercise has positive effects at patients with atherosclerotic cardiovascular diseases. These benefits are explained not only by the action upon the cardiovascular risk factors, but also by the improvement of the endothelial dysfunction. Physical exercise has vasodilator, antiplatelet, antioxidative, antiadhesive, antiproliferative and antiapoptotic effects upon the vascular endothelium. It improves the endothelium-mediated vasodilation in coronarian and peripheral beds in coronary heart disease, arterial hypertension, heart failure, diabetes mellitus, and also at sedentary and smokers, thus exerting a vasculoprotective action against atherosclerotic disease. PMID- 14756051 TI - [The prognostic molecular factors in precancerous lesions and nonmelanotic skin carcinoma]. AB - The most common cutaneous carcinoma, which includes basal cell carcinoma (BCC) and squamous cell carcinomas (SCC), accounts for approximately 90% or more of all skin malignancies. Actinic keratosis, bowenoid type of actinic keratosis, Bowen's disease or SCC in situ have been considered precursors of skin SCC. Authors present the histopathological criteria of precursors of skin SCC, variants invasive SCC and BCC. Also, one discusses about the roles of immunohistochemical staining of gene products in evidence of molecular changes in cell that might occur according to the transformation from precancerous lesions to SCC. PMID- 14756052 TI - [Oxidative stress in atherosclerosis]. AB - Oxidative stress is the result of the imbalance between pro-oxidant and antioxidant factors. The participation of oxidative stress in atherosclerosis (ATS) is reflected by lipid peroxidation which is a process initiated and maintained by oxygen reactive species generated by the aterogenic cells themselves. The endothelial aggression in ATS is accompanied by endothelial dysfunction, which is due to the neutralization of nitric oxide by superoxid anion. The key role in the onset and the development of the ATS lesions belongs to oxidated LDL-cholesterol that influence at different levels and by several mechanisms the ATS process. The neutralization of the toxic effects of free radicals is due to the endo and exogen antioxidant systems. It appears that the individual antioxidant status is influenced by environmental factors as well as by a genetic determinant. The antioxidant therapy, which is controversial at the moment, represents an associated therapy in endothelial dysfunction from ATS. PMID- 14756054 TI - [Pulmonary alveolar proteinosis]. AB - It is a disease of obscure cause that is characterized by the accumulation of a granular material that contains abundant lipid within the alveoli of lung. Pulmonary alveolar proteinosis (PAP) has been divided into a congenital and an adult form. The acquired form has been subdivided into a idiopathic form and a secondary form associated with a know disorder or exposure as silica, aluminium, titanium. Dyspnea and cough are the most common presenting symptoms. Chest pain, hemoptysis, fever and weight loss are variably reported. Pathogenesis remains unknown, but evidence points to a dysfunction of alveolar macrophages. Mice genetically deficient in granulocyte macrophagecolony stimulating factor (GM-CSF) show an alveolar proteinosis. A neutralizing antibody against GM-CSF was found in bronchoalveolar lavage fluid and serum of patients with idiopathic PAP. Currently, no specific therapy exists for pulmonary alveolar proteinosis, and sequential whole lung lavage is standard treatment. PMID- 14756053 TI - [Peripartum cardiomyopathy]. AB - Peripartum cardiomyopathy (CMP) represents an intriguing and incompletely characterised cause of heart failure arising in women without previously known heart disease during last trimester of pregnancy or first 20 weeks after birth. Fundamental clinical and basic research is lacking regarding this rare but potentially devastating disorder. The article reviews present accepted definition of CMP, epidemiological data, possible etiologic factors and pathogenic mechanisms proposed in CMP. It describes identified risk factors for CMP, clinical symptoms and signs, diagnostic assessment and treatment. Prognosis, follow up criteria and education for patients with previous CMP concerning subsequent pregnancies are also described. PMID- 14756055 TI - [Medical and social implication of cervical cancer screening]. AB - Invasive cervical cancer represents a major health problem in Romania. When there is a functioning screening network, the incidence of invasive lesions has a marked reduction, this program being the most successful cancer prevention program of all times. 80% of the actual incidence and mortality from this disease occurs in countries without an organized screening. A good infrastructure of the cytopathologist--practitioner relation must be achieved in order to reach the standard of early detection of the preinvasive cervical lesions. PMID- 14756056 TI - [Motivation for work based on needs--considerations from the health manager's viewpoint]. AB - Motivation represents the degree in which a persistent effort is focused to achieve a certain goal. Managers, including health managers, are especially interested about motivation, due to the direct relation with productivity. The paper presents two categories of motivations, extrinsic and intrinsic, from the point of view of individual's needs. This theory is known as Maslow's pyramid and shows that basic needs are on the first place in order of importance, followed by needs of safety, association and fulfillment. Satisfied needs cease to represent an effective motivation. Health managers must combine extrinsic and intrinsic motivations, in order to improve productivity. PMID- 14756057 TI - [Transcription regulation and coordination of some cell signaling genes in brain and heart of connexin 43 null mouse]. AB - We performed a cDNA microarray study of the transcription regulation and coordination of four gene families whose products are involved in cell-cell and cell-matrix interaction (ADAM, integrin, MMP, TNF) within brain and hearts of wildtype (WT) and connexin43 null (KO) neonatal C57Bl/6j mice. The study revealed that both WT brain and heart exhibit significant correlations among the transcriptions of cell-signaling genes and that depletion of Cx43 regulates both their expression and coordination. Adam8 was identified as the command gene of the group in WT and Mmp2 in KO, while Tnsf6 plays the dominant role in both WT and KO heart. Our results suggest that the functional coordination of cell signaling proteins may be related to the expression coordination of the corresponding genes presumably to ensure the efficiency of the functional pathways and that intercellular communication modulates cell-cell and cell-matrix interaction. PMID- 14756058 TI - [Loss of hormonal receptors expression from primary breast carcinoma to axillary lymph node metastases]. AB - Breast tumors growth is regulated by female sex steroid hormones. The level of the estrogen and progesterone receptors (ER and PR) expression by the malignant cells is important for the evaluation of the tumor prognostic and the benefit of a hormonal therapy. The aim of our study was to identify the expression of estrogen and progesterone receptors in primary breast tumors and in the corresponding axillary lymph nodes metastases, in 24 cases. The results showed that more than 30% of poorly differentiated breast carcinomas lost their expression of hormone receptors from the primary tumors to axillary metastases, an event which can be associated with an aggressive tumoral behaviour and resistance to hormonal therapy. PMID- 14756059 TI - [Fever of unknown origin: 1. Etiology. A prospective multicenter study of 164 patients]. AB - BACKGROUND: The spectrum of fever of unknown origin seems to be determined by geographic and economic factors, and it appears to change in time. Excepting a small retrospective study, no other study on fever of unknown origin has been performed in Central or Eastern Europe. METHODS: A multicenter cohort study was carried out on 164 consecutive patients who met the classic, modified criteria of fever of unknown origin. The study lasted 2 years (1997-1998) and included a follow-up period of another 2 years. MAIN OUTCOME MEASURED: The final diagnosis at the end of follow-up. RESULTS: 74 (45%) patients had infections (tuberculosis: 27 patients, 16%), 41 patients (25%) had neoplasms, 30 (18%) had non-infectious inflammatory diseases, three (2%) drug fever, and four (2%) other causes. The etiology remained obscure for 12 patients (7%). CONCLUSIONS: Infections represent the most important etiology, among them predominating tuberculosis. PMID- 14756060 TI - [Effect of antilipemic therapy on patients with diabetes mellitus and obesity]. AB - Fibrates are hypolipemic agents used in noninsulin dependent diabetes mellitus (NIDDM) because these drugs have no influence on glycemia control. The efficiency of fenofibrate (Lipanthyl) was studied in a group of 22 subjects, 30-70 years, obese (body mass index-BMI over 30 kg/m2) or overweight (BMI = 28-30 kg/m2) with hypertension or/and diabetes. All the patients received 200 mg fenofibrate daily, for 6 month, and they had a hypocaloric diet. We measured seric lipids. The results were compared with a control group (20 normal subjects). Under treatment, atherogenic lipids (total cholesterol, LDL-cholesterol and triglycerides) had significantly decreased (p < 0.05) and antiatherogenic fraction (HDL-cholesterol) significantly increased (p < 0.05) compared to initial values. In the mean time, the BMI has significantly decreased under fenofibrate treatment, this body mass loss having a positive influence on lipid metabolism. We observed a better therapeutically response in obese patients vs. the overweight ones, probably due to a higher prevalence of the hypertension in the second subgroup. In conclusion fenofibrate is the drug which can be choose in diabetic dyslipidemia without cardiac disease. PMID- 14756061 TI - [Early detection of diabetic nephropathy in children and teenagers with type I diabetes]. AB - Microalbuminuria (MA) is a noninvasive marker which has to be effected for all the patients in order to watch the appearance of the kidney disease. The aim of this study was the identification of the MA and of its associated factors within a lot of 110 children and teenagers suffering from type 1 diabetes mellitus. Persistent MA (the incipient diabetic nephropathy) was detected in the case of 10 patients (9%) and the intermittent one at 13 of them (11.8%). In all the cases, a poor glycemic control was associated, the average of the Hb A1c being higher than 10%. The progression of MA was associated with high blood-pressure values in the case of 2 patients and other chronic complications were present at 15 of them. In conclusion, we detected MA in 20.8% of the patients and its prevalence was significantly associated with poor glycemic control. PMID- 14756062 TI - [Electron microscopy in oral squamous cell carcinoma of elderly patients]. AB - OBJECTIVES: Squamous cell carcinoma is the most frequent tumor occurring in the oral cavity. The aim of the study is to point out the fine-structural characteristics of the tumor cells and of the adjacent stromal cells, trying to establish a correlation between these characteristics and the biological behavior of the tumor. MATERIAL AND METHODS: We performed an electron microscopic investigation for tree elderly patients diagnosed with oral squamous cell carcinoma. RESULTS: The tumoral cells presented nuclear abnormalities and the reduction of the intercellular cohesion, through an increase in the intercellular spaces, and desmosomal modifications. Through the abundance and the variety of the cellular types, the stromal component represented a positive response from the host. CONCLUSIONS: In the diagnosis of the oral carcinoma, the electron microscopic exam supplements the light microscopic investigation, revealing important morphologic elements at a subcellular level (regarding the nucleus, cellular organelles, and junctions). PMID- 14756063 TI - [Cutaneous and oral manifestations in HIV-infected children and adults--169 cases]. AB - The aim of the study was to evaluate cutaneous and oral manifestations in infected HIV patients. MATERIALS AND METHODS: We retrospectively analyzed 169 cases admitted in Infectiouse Disease Department of Iasi in 2001-2002 period. RESULTS: Cutaneous and oral manifestations were: candidiasis (99 cases), herpes virus infectious (36 cases), scabies and straphylococcal/streptococcal skin disease (26 cases), prurigo nodularis, psoriasis and verruca vulgaris (9 cases). Children of 0-13 year old group was 75.73 percent. Classification of HIV infection was related with CD4 count for 166 cases. Twelve cases with oral pharyngitis candidiasis, scabies and streptococcal skin diseases was 2-3 recurrent episodes of manifestations. Etiotrop treatment was associated with HAART therapy. CONCLUSIONS: Cutaneous and oral manifestations are occurred frequently in HIV infected patients, with a various etiology, but the severity, persistence and its evolution did not evaluate. PMID- 14756064 TI - [Atrio-ventricular canal: possibilities of recovery]. AB - The evolution and prognosis of the atrioventricular canal (AVC) depend very much on the clinical type of the disease and its recovery is sometimes an extremely difficult problem. METHODS: The evolution of the 24 cases with atrioventricular canal (10 boys and 14 girls), registered in the files of pediatric cardiology consulting room from the Outpatient Department of the "Sf. Spiridon" Hospital from Iasi, has been studied. According to the state of the patients and, especially, with the parents' permission, these children were sent to Cardiovascular Surgery, for corrective surgical treatment. Before that, they had received a medical treatment for the cardiac failure (digitals, diuretics) and for the pulmonary hypertension (beta-adrenergic blockers, renin angiotensin inhibitors) and they continued this treatment for 1-2 years after the surgical correction. RESULTS AND DISCUSSIONS: This sample included: 9 cases with AVC complete type; 1 case with AVC-intermediate type; 14 cases with AVC-partial type (13 cases with atrial septal defect-ostium primum + mitral valve cleft and 1 case with left ventricle to right atrium type IIa communication). Only 7 of the 10 children with AVC partial type were surgically corrected, and in all cases the evolution was positive. Three of the nine cases with AVC complete type were investigated in Cardiovascular Surgery in our country, all of them considered without surgical solution. For one of the cases with AVC--complete type and pulmonary stenosis, considered also without surgical solution, a palliative systemic-to-pulmonary artery shunt was made, for the improvement of the pulmonary circulation. CONCLUSIONS: 1. AVC partial type could be recovered in the best conditions in the clinics of cardiovascular surgery in our country. 2. AVC- complete type didn't have, in our cases, surgical solution, either in our country or abroad. 3. For the special situation of the AVC complete type with pulmonary stenosis, the only treatment possible was palliative, performed to increase the pulmonary artery blood flow. PMID- 14756065 TI - [Determination of urinary iodine as a marker of iodine deficiency]. AB - PURPOSE: Determination of ioduria in order to establish the actual degree of iodine deficiency in school-age children from 4 counties in Moldavia. MATERIALS AND METHODS: Urinary iodine was determined in 927 school-children aged 7-14 years from 4 counties: Suceava, Iasi, Neamt and Botosani. The determination was made by cerium-arsenate method and by a semi-quantitative method. RESULTS: The median value of ioduria was 3 micrograms/dL in Suceava, 8.8 micrograms/dL in Iasi, 7.1 micrograms/dL in Botosani. In Roman-Neamt 93.75% of all samples had a value of ioduria below 10 micrograms/dL. From this point of view, Suceava and Roman are with moderate iodine deficiency; Iasi and Botosani are with mild iodine deficiency. CONCLUSIONS: Despite the existence of a program of iodine prophylaxis, there are still many regions in Moldavia with iodine deficiency, so it is important to have more measures of sanitary education and prophylaxis in these regions. PMID- 14756066 TI - [Survival in a cohort of patients with rectal cancer]. AB - BACKGROUND: The aim of our study is to analyze the five-year survival for patients with operated rectal cancer in our clinic during a five-year period (1997-2001). METHODS: Our epidemiological clinical study is based on prospective analysis of 196 cases of rectal cancer operated between 1997 and 2001. Survival evaluation was prospective cohort follow-up study. We have used Epiinfo 2000 software package for data analysis. RESULTS: The five-year survival was 54.08% if we take into account all deceases including other causes and 64.28% for exclusive deaths caused by rectal cancer and metastasis. The survival at 5 years (S5) was: stage A S5 = 85.3%; stage B1 S5 = 76.6%; stage B2 S5 = 71%; stage C S5 = 68.7% and stage D S5 = 8.6%. The follow-up in month (e5) was: stage A e5 = 58, stage B1 e5 = 46; stage B2 e5 = 36, stage C e5 = 20 and stage D e5 = 6. The values for S5 and e5 are in accordance with the literature. CONCLUSIONS: Despite progresses of rectal cancer management, the five-year survival doesn't transcend much over 50%. Because the five-year survival for A stage is 85.3% in our study, we may conclude that at present introducing of screening program represents the only chance for improvement of results. PMID- 14756067 TI - [In vitro antibacterial activity and beta-lactamase stability of meropenem]. AB - The comparative activity of meropenem with that of imipenem, cefotaxime, ceftriaxone and ceftazidime against 856 gram-negative bacilli was studied studied by an agar dilution method. Meropenem and imipenem were high active against tested strains. Resistance to third generation cephalosporins was high for most microorganisms tested. For rapid detection of metallo-beta-lactamase (MBL) producing gram-negative bacilli a simple disk diffusion test was used. EDTA, FeCl2 and CuCl2 were evaluated as IMP-1 inhibitors. The method is helpful for screening of IMP-1 producers in daily clinical activity. PMID- 14756068 TI - [Activity of fourth generation cephalosporins against clinical isolates of Enterobacteriaceae]. AB - Cefpirome and cefepime are a novel group of cephalosporins which contain a positively charged quaternary ammonium at carbon 3 of the dihidrothiazone ring. The antimicrobial agents cefpirome, cefepime, cefotaxime, ceftriaxone, ceftazidime, cefoperazone and imipenem were tested against clinical isolates of Escherichia coli (n = 302) and Klebsiella spp. (n = 62) obtained during september december 2002 from patients of Galati Emergency Hospital. The fourth generation cephalosporins cefpirome and cefepime have similar in vitro activities to the third generation cephalosporins. E. coli showed the comparable resistance rates for all cephalosporins. Against Klebsiella spp. strains cefpirome was less active (35.5% resistance) than cefepime (25.8% resistance). As expected, imipenem had excellent activity (100% susceptibility). PMID- 14756069 TI - [Reasons for conservative treatment in breast cancer]. AB - We analyze 112 patients with breast conservative treatment operated in our clinic in the last 5 years. Breast preservation surgery represented 30.35% of all cases treated for breast cancer in this interval of time. In other 63 situations conservative treatment was attempted but finally given up due to justified motivations and decision for mastectomy was adopted as optimal. Rigorous selection of patients for conservative surgery was our care. Tumor size, excisional margins, nodal status were the main criteria. Correlation with breast volume, age, patient's opinion were also important. Preservation of breast in women interested and cosmetic results were the purposes of our attitude. Breast conserving therapy may be indicated only respecting the oncological principles and close surveillance of patients is obligatory. PMID- 14756070 TI - [The contribution of cystography in the diagnosis of bladder trauma]. AB - With the aim of the evaluation of cystography as a diagnostic method, this study has retrospectively analysed 22 cases of bladder rupture (19) or perforation (3). When performed, the retrograde cystography has made the accurate diagnosis immediately in all the 15 cases (100%). The adequate filling and the x-rays made after the bladder evacuation are the main conditions, that allow the visualisation of the contrast material extravasation. Out of the remaining 7 cases, the urographic cystography has made the correct diagnosis in 6 of them (86%) and the method should be indicated in cases with concomitant rupture of the membranous urethra. PMID- 14756071 TI - [Antibiotic prophylaxis in surgery for colorectal cancer]. AB - The prophylactic use of antibiotics in the purpose of decreasing the frequency and severity of surgical infections is still controversial. The practical need of defining the concept of antibiotic prophylaxis gave rise to numerous polemics in the literature. This paper presents a retrospective study on 103 cases whom diagnostics were colon and rectum cancer, that were operated in the IV-th Surgical Clinic in 1993-2002 period; at these patients was done antibiotic prophylaxis local and systemic before, during and after operating. This method made possible the registration of a minimum number of cases with surgical infection (7 cases, representing only 7% of the operated patients): a case (1%) with peritoneal collection (needing the surgical evacuation of this collection) and in 6 cases, parietal superficial infections, which imposed the prolong of drainage for almost 7 days. Starting from the basic principles of antibiotic therapy, this paper aims at outlining practical guidelines for a judicious antibiotic prophylaxis. PMID- 14756072 TI - [Role of computerized tomography in the diagnosis of transitional cell carcinoma of the upper urinary tract]. AB - PURPOSE: To evaluate the computed tomography (CT) semiological criteria used in the diagnosis of transitional cell carcinoma of the upper urinary tract. MATERIALS AND METHODS: 26 patients with transitional cell carcinoma of the upper urinary tract underwent preoperative CT. In all cases nephroureterectomy was performed and CT and histopathologic findings were compared. RESULTS: 16 tumors were infiltrative and 10 tumors were sessile. The smallest tumour had 7 mm diameter and the largest one 13 cm. Associated focal hydronephrosis appeared in 11 cases and diffuse hydronephrosis in 7 cases. The vast majority of these tumors (69.23%) had a 21 to 50 Hounsfield Units enhancement. CONCLUSIONS: CT is a powerful tool to diagnose transitional cell carcinoma of the upper urinary tract when data acquired in intravenous urography and echography are equivocal. PMID- 14756073 TI - [Assessment of food intake in rural area school children]. AB - Children food intake is influenced by family nutritional habits and economic status. According to the results of our survey, only 54.8% of the children have taken their breakfast, although all of them recognize the importance of this meal. 9.6% of the children are not using fresh fruits and vegetables in their diets, even if these are the main sources of vitamin C. Regarding the milk consumption, most of the children use this food item once (35.5%) or twice (19.4%) a day and only 16.1% occasionally. Although most of the children (93.5%) know that excessive salt intake represents a risk factor for their health, 87.1% have preferences for salted and spiced foods. Under the influence of advertising, certain changes in the nutritional habits have been noticed; instead of traditional snacks, children are more attracted to fast-food type snacks. PMID- 14756074 TI - [Effect of environmental factors on high school children's health ]. AB - We studied 78 teenagers from Iasi Pedagogic High School. Body mass index values are mainly (83.3%) normal. 60.3% of the teenagers are coming from rural areas, but there are no significant differences between children coming from urban and rural areas. The frequency of families with more than 5 children in the urban areas is very small (1.3%), but in the rural areas this frequency is more than 10 times higher (15.4%). Due to this factor, the per capita income is reduced in the rural areas. Nutrition is a major problem in both urban and rural areas, due to unbalances food intake. PMID- 14756075 TI - The use of the supra-pubic fibrous complex in isthmic hysteropexies. AB - This study forwards a new surgical approach we have performed in the treatment of genital prolapse, associated or not with stress urinary incontinence. The study used 150 patients with genital prolapse and stress urinary incontinence in various stages, and lasted from 1985 until 2001. Four-five threads of silk were passed through the uterine muscle in the isthmus area, just below the bladder uterus recess, and fastened to the supra-pubic ligament complex. Although the mechanical basis for this surgical approach is not entirely clear, no relapse, incident or post-surgery complications (bladder voiding problems, enteroceles, detrusor instability, urethra or bladder bottom lesions, incomplete urinary retention or clearance, hematoma or abscess of the Retzius space) were noted in any of the studied cases. The suspension of the uterine isthmus on to the supra symphyseal fibrous complex leads to an increase in urethral closure pressure during the occasional increase of intra-abdominal pressure (cough, laughter, spontaneous movements, walking), the results being superior to the other methods used for treating genital prolapse. PMID- 14756076 TI - [Celiac disease associated with Helicobacter pylori infection]. AB - Celiac disease, also known as gluten-sensitive enteropathy, is an autoimmune enteropathy caused by the ingestion of gluten-containing grains in susceptible subjects. The authors present a 3 years and 5 months old girl diagnosed with celiac disease at 1 year and 5 months old. Initially, the evolution after gluten free diet was favorable. After 2 years the child presented abdominal pain and anorexia. The IgA antigliadin antibodies had normal values. The gastric biopsy found Helicobacter pylori gastritis. After treatment for Helicobacter pylori eradication the symptoms disappeared. PMID- 14756077 TI - Atypical post-renal transplantation hyperparathyroidism--further support for "removing all enlarged glands". AB - Hyperparathyroidism is an important sequela of chronic renal failure, remains a considerable challenge to nephrologists, and can be seen as inevitable in patients undergoing long-term renal replacement therapy. As time with renal disease increases then so does the cumulative risk of hyperparathyroidism, and the eventual need for surgical parathyroidectomy when hyperparathyroidism becomes refractory to medical intervention. Parathyroidectomy before dialysis treatment has started, or after successful renal transplantation, is much less commonly performed than when the patient is receiving dialysis. Increasingly the propensity for residual parathyroid tissue left behind (by design or accident) at an initial parathyroidectomy to undergo progressive hyperplasia under the constant stimulus of uremia, and by so doing result in the need for a second, more complex, neck exploration, has increased support for initial total parathyroidectomy for patients on dialysis. The optimal operative procedure for autonomous hyperparathyroidism after successful renal engraftment is however less clearly established. We discuss two very unusual but instructive cases of post renal transplantation autonomous hyperparathyroidism requiring surgical parathyroidectomy. Using these cases as examples we discuss the various surgical options, and discuss the contentious issue of the place for autografting parathyroid tissue. PMID- 14756078 TI - [Jejunoileal bypass]. AB - Obesity is a condition which can be found very frequently today, both in developed and 3rd world countries. The incidence of obesity in adult population of Romania is about 35%, and most of these patients are females. We'll present the case of a 54 years old woman with BMI = 57 kg/m2, who was hospitalized for the treatment of a postoperative eventration after an umbilical hernia. Her nocturnal breathing troubles, knee pains and walking difficulties made us consider the idea of a digestive by-pass. The surgical intervention consisted of jejunoileal by-pass, abdominoplasty and dermolipectomy with bipolar drainage. Many complications occurred in the postoperative period (renal failure due to severe diarrhea). The weight loss after 18 months was 37%, which means 66% of the weight surplus (similar results can be found in professional statistics--around 70%). After 18 mounts her weight is 95 kg and she allowed to consume any food. 18 mounts after the operation, the number of stools decreased to normal (1-2 per day). In conclusion the morbid obesity can and must be treated surgically. Jejunoileal by-pass is a highly effective procedure, but surgeons must be aware of the pact that severe complications which may occur anytime and must be treated immediately. After this kind of operation, weight stabilization can be achieved within 2 years, no diet being necessary as an additional treatment. PMID- 14756079 TI - [Morphea associated with toxoplasmosis]. AB - Circumscribed scleroderma or morphea is a rare disease that involves limited areas of skin and usually is not associated with visceral lesions. Its etiology is still debated, but its appearance signifies a particular immunological status. We present a case of a male patient, which developed morphea lesions during the infection with Toxoplasma gondii. Clinical manifestation was characteristic for the classic plaque-type of morphea. Diagnosis was confirmed by the histopathologic examine of the cutaneous lesions. The evolution of lesions was correlated with antibodies titre for Toxoplasma gondii, and local administration of corticosteroids accelerated the evolution to cutaneous lesions stabilization. PMID- 14756080 TI - [Malignant mixed mesodermal tumor: histo- and cytopathologic correlations]. AB - To correlate the cytopathological and the histopathological findings in uterine mixed mesodermal malignant tumor (MMMT) we have examined the cervical smear, endometrial curettage and hysterectomy specimen of a patient diagnosed with uterine tumor. The smear was stained by Papanicolaou staining and the tissue processed by routine technique and stained H&E. The original cytological diagnosis was adenosquamous carcinoma. The histopathological diagnosis was MMMT of heterologous type. A review of the smear revealed features which may orientate the diagnosis: multinucleate cells, isolated cells with cyanophilic cytoplasm, hyperchromatic nuclei and prominent nucleoli, elongated cyanophilic cells of sarcomatous origin. We conclude that the cytopathological diagnosis of the MMMT in cervical smears is very difficult. This may be sustained by the evidence of more cell types and cellular features orientating to a sarcomatous origin. The most important differential diagnosis is adenosquamous carcinoma. PMID- 14756081 TI - [Endometrial carcinoma in situ. Morphological and immunohistochemical aspects]. AB - A possible precursor for invasive endometrial carcinoma is an in situ stage, but this notion is very much debated. We present a case of endometrial carcinoma diagnosed in a patient with post-menopausal bleeding. The morphological examination of the hysterectomy specimen, processed by routine technique, put in evidence focal atypias of the glandular epithelium in a group of few endometrial glands, on a background of atrophy without a desmoplastic stromal reaction. The immunohistochemical investigation of the steroid receptors and p53 protein, using monoclonal antibodies in the ABC technique, revealed the p53 positivity and the absence of the receptors in the focal malignant areas. The surface epithelium had the same immunohistochemical aspect. These aspects were decisive in precising that the malignant change was primarily in the surface epithelium with secondary extension in some glands, the final diagnosis being in situ serous papillary carcinoma. PMID- 14756082 TI - [Video-stroboscopy, a method for exploration of laryngeal pathology]. AB - The authors present the importance of stroboscopy in the larynx examination. They insist to prove the functional modifies of the voice specially at the singers also all patients who use the voice more frequently. This method permits to make a differential diagnosis between the inflammatory and tumoral diseases of the larynx. PMID- 14756083 TI - [The polarimetric determination of pentavalent vanadium by using quinine chlorohydrate as reagent]. AB - The researches of the authors on the quinine chlorohydrate QH+Cl- and sodium metavanadate (Vv) reaction have led to the elaboration of two methods--a conductometric one and a gravimetric one--of dozing the QH+Cl- and, in general, of any quinine soluble salt with NaVO3. The methods, that have applications in the domain of drugs and is based on the formation of a white precipitate, in which the molar ratio QH+:Vv = 1:1. Since some authors have used quinine as a reagent in order to determine polarimetrically some ions (Ag+ and Cu2+), by taking into consideration the optic activity of quinine, we have aimed at using the formation reaction of QHVO3 for quantitatively determine the Vv with quinine chlorohydrate, by measuring the rotation angle of polarized light alpha both for the excess of QHCl reagent and for the solution obtained by dissolving the QHVO3 precipitate formed within the reaction. For this purpose there has been necessary the construction of two standard scales alpha = f(c) and there has been suggested a working methodology for the dozing of Vv with quinine chlorohydrate or any other of its soluble salts, by using any laboratory polarimeter. The results are satisfactory and depend on the sensitivity of the apparatus. PMID- 14756084 TI - [UV spectrophotometry used for quantitative determination of oleanolic acid]. AB - In order to adapt the UV spectrophotometry method to oleanolic acid dosing, absorption spectra within the range from 230 to 350 nm have been determined for both OA systems in glacial acetic acid (lambda max = 256 nm) and OA systems in ethyl alcohol (lambda max = 277 nm). The time stability of oleanolic acid in the presence of the two solvents has also been investigated in this study. Besides, the composition of etalon series and graphical representation of experimental results (n = 6) have been performed. Also, the calculation algorithm was given, the regression lines have been drawn and statistical parameters were calculated to establish the mathematical expression of OA concentration (mg%). PMID- 14756085 TI - [Assistant professor Dr. Gabriel Marderos (1923-2003)]. PMID- 14756086 TI - [Old-fashioned childbirth alternatives challenge modern obstetrics. Home childbirth and doula support controversial]. PMID- 14756087 TI - [Compensatory gene mechanisms can take over in cancer]. PMID- 14756088 TI - [Doula--a new concept in obstetrics]. AB - We wanted to study the effect of extra emotional support in the form of a non professional woman (doula) before and during delivery. About 200 primiparae were invited to participate in a prospective study which intended to assess differences in delivery outcome between women with and without a doula. Fifty four declined to participate, 55 had a delivery with doula and 46 were controls. Lower rate of emergency caesarean sections in the doula-group was noted. The parents as well as the staff, became to regard the doula as a valuable support during delivery. PMID- 14756089 TI - [The county of Stockholm pays for home childbirth in certain cases. Strict medical criteria and requirements when it comes to midwives must be fulfilled]. PMID- 14756090 TI - [Functional MRI of the elderly can complete the picture of cerebral functions]. AB - Functional Magnetic Resonance Imaging, fMRI, is a method for the study of neuronal circuits underlying cognitive, sensory, emotional and motor activities in the living human brain. An important question is whether elderly persons and patients with cognitive deficits differ from others as regards neuronal activation. Not many studies have been published but some of them have shown surprising results. This new research has shown some very interesting new facts about the vascular response and the regulation of the microvasculature in the brain. Most fMRI studies have been performed on healthy young males, but in our opinion this method could reveal new findings to complete the picture of human cognition if studies were to be performed involving elderly people and patients with dementia. PMID- 14756091 TI - [Remote ophthalmologic examination as a new cooperation between a specialist and a community health center]. PMID- 14756092 TI - [Experiences with telemedicine in psychiatric care in the county of Vasterbotten. A practical working tool which is used more and more]. PMID- 14756093 TI - [Rehabilitation after stroke. Imaging techniques show how the cortical reorganization is affected by training]. AB - It is widely accepted that reorganisation of the brain occurs after a focal brain lesion such as stroke. Neuroimaging methods are used to study such reorganisation in vivo. Improvements in arm and hand motor function during recovery post-stroke have been related to reorganisation in primary and secondary sensorimotor areas by indirect measure of synaptic activity with functional MRI. Reorganisation occurs in both the affected and the unaffected hemispheres. Preliminary training studies post-stroke have shown correlations between improvements in motor function and brain activity changes. Recent research findings are reviewed herein. Further understanding of the neurobiological mechanisms of post-stroke recovery will lead to development of optimal treatment strategies during rehabilitation of stroke survivors. PMID- 14756094 TI - [Geographical inequalities in the distribution of general practitioners in Sweden]. AB - AIM: To estimate trends in the inequalities in the distribution of general practitioners in Sweden during the past fifteen years. METHODS: Relative inequality indicators. We plotted the Lorenz curves and calculated the Gini coefficients for the entire country during 1986, 1991, 1996 and 2001. RESULTS: During the past 15 years in all a declining trend was noted in the inequalities in the geographical distribution of general practitioners in Sweden, with the lowest level in 1996. CONCLUSIONS: The degree of inequality in the geographical distribution of general practitioners appeared rather stable during the period, although with some increase in the final year studied. Since equality is part of quality, policy makers in all levels--national, regional and local--should pay attention as to how human resources are distributed. PMID- 14756095 TI - [Physicians' travels abroad supported by drug industry scrutinized. Three cases surveyed by the IGM and the NBL]. PMID- 14756096 TI - [Time to proceed towards better quality in health care. Lessons learned from USA and England give good support]. PMID- 14756097 TI - [Medical documentation through the ages]. PMID- 14756098 TI - [Medicine and poetry. Scientific language is not enough]. PMID- 14756099 TI - ["Beware of physicians! Remember they all are sales agents and must live! August Strindberg's relations to physicians and his paranoid psychosis]. PMID- 14756100 TI - [A scandal in the society, wrong diagnosis. Lady Flora (not pregnant) died because of peritonitis]. PMID- 14756101 TI - ["Compulsory referral" benefits the patients. An interesting initiative in the county of Stockholm]. PMID- 14756102 TI - [Doxazosin and heart failure: a clinical conclusion is wanted]. PMID- 14756103 TI - [Alfadil--positive documentation when it comes to safety is missing]. PMID- 14756104 TI - [Unwanted "psychiatrism" in psychiatry]. PMID- 14756105 TI - [Establish a national unit and change legislation concerning persons with brain injury and substance abuse]. PMID- 14756106 TI - [An excellent idea to solve health care economics]. PMID- 14756107 TI - [Undergraduate medical education should be pursued close to patients with "common" complaints]. PMID- 14756108 TI - [6000 crown-limit of the "free pass" can solve the crisis of health care in the county of Stockholm]. PMID- 14756109 TI - [Shadowy postgraduate students haunt the National Agency for Higher Education]. PMID- 14756111 TI - Health tips. Helping others hear. PMID- 14756110 TI - Corticosteroids. An evolving story. PMID- 14756112 TI - Treatment for chronic sinus infection studies at Mayo Clinic. PMID- 14756113 TI - Flu shots for heart disease and stroke protection. PMID- 14756114 TI - Root canals. Tooth-saving repairs. PMID- 14756115 TI - Yoga and breathing. Continuous, rhythmic. PMID- 14756116 TI - Blood pressure. Defining a new normal. PMID- 14756118 TI - We try to keep small objects away from our toddler grandson. However, we still worry about him accidentally swallowing something. What should we do if this happens? PMID- 14756117 TI - I've heart that coral calcium is one of the best-absorbed calcium sources because it's natural. I'm thinking about switching from my usual calcium citrate supplement. PMID- 14756119 TI - Sympathetic nervous system as transmitter of mechanical loading in bone. AB - Sympathetic innervation has been demonstrated in bone. Adrenergic stimulation is one of the transmitters of bone loss by uncoupling between decreased bone formation and increased bone resorption. OBJECTIVE: By using a non-specific antagonist of -adrenergic pathway (propranolol per os), we hypothesized that we could rescue the uncoupling induced mechanical unloading bone loss in the rat model of tail-suspension. MATERIALS AND METHODS: Twenty-two female Wistar rats, 12 week-old, have been divided into three groups: eight tail-suspended rats (SR), six tail-suspended rats treated by propranolol (SRP) and eight non-suspended rats (NSR) during 30 days. Bone mineral density (BMD, g/cm2) has been measured by DXA (Hologic QDR-4500A) at D0 and D30 of the study, in the distal femoral metaphysis (DFM), the femoral diaphysis (FD), the whole body (WB, g) and body composition. RESULTS: Between D0 and D30, in DFM a significant variation in BMD is observed between NSR and SR (% BMD change: NSR +15.6 +/- 3.1% vs SR -1.0 +/- 1.4%, P < 0.0001) and BMD rescue in SRP group (% BMD change SRP +5.3 +/- 1.5% vs SR -1.0 +/ 1.4%, P = 0.03). In FD, gain of BMD is significant in NSR compared to SR (+17.5 +/- 1.5% vs +8.2 +/- 2.8%, P = 0.007) and to SRP (+17.5 +/- 1.5% vs +10.1 +/- 2.4%, P = 0.046). Gain in SRP group is not significant compared to SR group (P = 0.6). In WB, SRP gain more BMD than NSR (+14.0 +/- 1.8% vs +5.4 +/- 0.7%, P = 0.0002) and than SR (+14.0 +/- 1.8% vs +7.8 +/- 1.4%, P = 0.0043). There is no difference between NSR and SR groups (P = 0.19). CONCLUSION: We demonstrate that adrenergic pathway of sympathetic nervous system is a major transmitter pathway of mechanical loading in rat bone. A specific study is necessary to analyse a possible systemic effect of propranolol in rat bone. Propranolol could be used to prevent the induced mechanical unloading bone loss as weightlessness PMID- 14756120 TI - Longitudinal femoral shaft due to bone insufficiency. A review of three cases. AB - We report three new cases of longitudinal femoral shaft fracture due to bone insufficiency and review the eight cases reported in the literature. The typical patient is a woman older than 65 years of age who present with mechanical pain in the thigh and/or groin. Palpation of the thigh may reproduce the pain. The diagnosis is often made late because the radiographs are normal initially. However, an early and consistent finding is increased radionuclide uptake along the femoral shaft. The fracture line is readily evidenced by computed tomography but may be difficult to see on magnetic resonance imaging. Use of crutches for 6 weeks to protect the bone from weight bearing ensures healing of the fracture. PMID- 14756121 TI - Half a century since the clinical introduction of chlorpromazine and the birth of modern psychopharmacology. PMID- 14756122 TI - Subcutaneous exophiala jeanselmei infection in a heart transplant patient. PMID- 14756123 TI - Health and environmental impact of nanotechnology: toxicological assessment of manufactured nanoparticles. PMID- 14756124 TI - Aldosterone antagonism and arterial stiffness. PMID- 14756125 TI - Dolichol: an essential part in the antioxidant machinery of cell membranes? PMID- 14756126 TI - Progress in development of a live-attenuated, tetravalent dengue virus vaccine by the United States Army Medical Research and Materiel Command. PMID- 14756127 TI - Coming changes in veterinary anatomy: what is or should be expected? PMID- 14756128 TI - Vitamin C in heart failure: hype or hope? PMID- 14756129 TI - [Ultrasound of the kidney and renal vessels. I: Normal findings, congenital diseases, diseases of the kidney parenchyma]. AB - Renal ultrasonography has become the standard imaging modality in the investigation of kidneys because it displays excellent anatomic detail, requires no special preparation of the patient and does not expose the patient to radiation or contrast agents. Ultrasonography is used to determine the site and size of the kidney and to detect focal lesions like tumors, cysts and renal stones. Furthermore the presence and urodynamic relevance of hydronephrosis can reliably be revealed. Also renoparenchymatous diseases are discernible to the experienced investigator, however most glomerular diseases cannot be further subclassified. Exceptions are primarily renovascular disorders like hypertensive nephrosclerosis, diabetic nephropathy or renal vasculitis. Color Doppler sonography allows the detection and quantification of renal artery stenosis, increased resistance index values may indicate irreversible disease. Ultrasonography has also been found of value in the evaluation of renal transplant kidneys. Especially in the early transplant course potentially fatal but reversible diseases like renal vein thrombosis or urinomas are detected with high sensitivity. In the long term, an increased resistance index value may also predict allograft failure. PMID- 14756130 TI - Antioxidant activities and oxidative stress byproducts in human hypertension. PMID- 14756131 TI - The traffic in Victorian bodies: Medicine, literature, and history. PMID- 14756132 TI - Current awareness in human psychopharmacology. PMID- 14756133 TI - Andrew Duncan and the health of nations. PMID- 14756134 TI - [Diagnosis and treatment of anterior and posterior shoulder dislocation. II. Treatment]. AB - In Part I the anatomy, biomechanics, different pathologic changes, and diagnostic tools for anterior and posterior instability were discussed. In this second part, treatment of an acute dislocation is introduced. Furthermore different surgical procedures and their indications are discussed. Algorithms for treatment of first time dislocation, chronic anterior instability, and posterior instability are presented. PMID- 14756135 TI - Employment trends in 2003. PMID- 14756136 TI - Bringing a dose of science to the art of medicine. PMID- 14756137 TI - FDA guidance on pharmacogenomics data submission. PMID- 14756138 TI - Antidepressant use in children questioned. PMID- 14756139 TI - 'New' FDA delivers in 2003. PMID- 14756140 TI - Daptomycin. PMID- 14756141 TI - Pursuing a career in medical sales. PMID- 14756142 TI - The conductorless orchestra. PMID- 14756143 TI - Lysosomal storage diseases market. PMID- 14756144 TI - Oxaliplatin. PMID- 14756145 TI - Basaloid cell carcinoma of the prostate. AB - We report on a case of malignant basal cell carcinoma of the prostate combined with a poorly differentiated conventional adenocarcinoma, infiltrating bilaterally the seminal vesicles and with lymph-node and distant metastases. Basal cell carcinoma represented the prevalent component of the tumour (80%), showed high mitotic activity, extensive perineural infiltration and vascular invasion and was immunoreactive for basal cell related antigens (high molecular weight cytokeratins 34betaE12 and p63). Our data confirm previous observations that basal cell carcinoma represents aggressive tumour with an adverse clinical outcome. PMID- 14756146 TI - Neuroendocrine differentiation in hyalinizing trabecular tumor of the thyroid. AB - The presence of neuroendocrine cells is putative in thyroid hyalinizing trabecular tumor (HTT), although this entity requires a differential diagnosis from paraganglioma or medullary carcinoma of the thyroid, due to similarity of the growth pattern of the cells. Here, we present a case of HTT with a mixture of endocrine cells stained positive for somatostatin, chromogranin A and Grimelius' silver impregnation. The histology and results of other immunostainings were consistent with the features typical for HTT. In the world literature to date, only two cases of HTT with endocrine cells, including the current case, have been reported. Nevertheless, the cases may indicate the diversified differentiation of cells in HTT and may, in part, account for the resemblance of HTT to paraganglioma and medullary carcinoma. PMID- 14756147 TI - Sinus histiocytosis with massive lymphadenopathy Rosai-Dorfman: three unusual manifestations. AB - We report on three exceptional courses of sinus histiocytosis Rosai-Dorfman. Patient one developed regional lymph-node disease subsequent to two independent malignancies in the right head and neck region. Patient two suffered from extensive extranodal disease with more than 100 mucocutaneous lesions over 17 years, which spontaneously resolved. Patient three showed exclusively extranodal disease, including bilateral conjunctival/scleral lesions, before he developed lung cancer. Our cases are unique for three reasons: the association of the disease with solid malignancies in two cases, the extent and persistence of exclusively extranodal disease in one patient and the appearance of thus far undescribed conjunctival/scleral lesions. PMID- 14756148 TI - Saccadic oscillations facilitate ocular perfusion from the avian pecten. AB - THE evolution of the eye is constrained by two conflicting requirements--good vascular perfusion of the retina, and an optical path through the retina that is unobstructed by blood vessels. Birds are interesting in that they have higher metabolic rates and thicker retinas than mammals, but have no retinal blood vessels. Nutrients and oxygen must thus reach the neurons of the inner retina either from the choroid through 300 micron of metabolically very active retina, or from the pecten, a pleated vascular structure protruding from the head of the optic nerve into the vitreous chamber, and more than a centimetre away from some retinal neurons. Despite the diffusional distance involved, several lines of evidence indicate that the pecten is the primary source of nutrients for the inner retina: the presence of an oxygen gradient from pecten to retina, the large surface area produced by macroscopic folds and by microscopic infoldings of the luminal and external surfaces of the capillary endothelium, extrusion of circulating fluorescein, high content of carbonic anhydrase and alkaline phosphatase, and retinal impairments after pecten ablation. Another peculiarity of birds, their saccadic oscillations, occur with a large cyclotor-sional component during every saccadic eye movement. In different species, saccades, which occur at intervals of 0.5-40 s, have up to 13 oscillations with frequencies of 15-30 Hz and amplia-tudes of about 10 degrees. Therefore, as much as 12% of some birds' total viewing time may be subject to the image instability caused by the oscillations. Using fluorescein angiography, we show here that during every saccade, the pecten acts as an agitator which propels perfusate towards the central retina much more effectively than is observed during intersaccadic intervals. PMID- 14756150 TI - Bibliography. Current world literature. Immunotherapy. PMID- 14756149 TI - Bibliography. Current world literature. Primary immune deficiency disease. PMID- 14756151 TI - AHA report says hospital profits are up for the first time since 1996. With admissions, ER visits and number of uninsured on the rise, acute-care facilities have to work harder. PMID- 14756152 TI - Is there something fishy about GM pets? PMID- 14756153 TI - Pig news in transgenics. PMID- 14756154 TI - Dealing with IACUC dysfunction. PMID- 14756155 TI - The green anole (Anolis carolinensis): a reptilian model for laboratory studies of reproductive morphology and behavior. AB - The green anol (Anolis carolinensis) is an excellent reptilian model for studying reproductive behavior and the neural and muscular morphology that supports it. This lizard has been the subject of behavioral and ecological study for more than 100 yr, and a rich literature exists on its natural history. Both courtship and copulatory behaviors reveal sex and seasonal differences, which allow for the study of mechanisms regulation naturally occurring variation in performance at multiple levels within a single animal model. Green anoles are readily obtained due to their abundance in the wild; once in the laboratory, they are easily maintained, bred, and reared. Background on the natural history and husbandry of this lizard is provided, and the authors' research program on the regulation of reproductive anatomy and behavior is reviewed, Discussion includes the similarities and differences in the mechanisms mediating both structure and function compared with more traditional animal models. This type of comparative research will make it possible to identify the fundamental principles governing reproductive biology, thus advancing both basic and applied knowledge. PMID- 14756156 TI - The male red-sided garter snake (Thamnophis sirtalis parietalis): reproductive pattern and behavior. AB - Among the small group of species (e.g., some temperate zone turtles, snakes, and bats) that exhibit a dissociated reproductive pattern, the red-sided garter snake (Thamnophis sirtalis parietalis) is probably the most well studied. For these species, courtship and mating occur immediately upon emergence from winter dormancy; the gonads remain essentially inactive. Male red-sided garter snakes are a particularly informative animal model for examining the role of neuroendocrine factors associated with reproductive physiology and behavior because unlike species that exhibit an associated reproductive pattern, in which sex steroids initiate and control sexual behavior, reproductive behavior in the male garter snake appears to be independent of circulating sex hormone control. In fact, the only factor associated with the initiation of courtship behavior and mating in the male garter snake is an extended period of low temperature dormancy followed by exposure to warm temperatures. Yet the presence of sex steroid concentrating neurons within the pathways regulating courtship and mating suggests that sex hormones may be involved in the activation of sexual behavior. Although circulating androgens are elevated upon emergence from hibernation, the initiation of courtship behavior and mating appears to be independent of direct androgen control. Thus steroid hormones may have indirect effects on mating behavior in animals that display "dissociated" reproductive behaviors. PMID- 14756157 TI - The assessment of quality of life in dementia. PMID- 14756158 TI - The meaning of words. PMID- 14756159 TI - Toward an early diagnosis and treatment of Alzheimer's disease. AB - Alzheimer's disease (AD) is the most common neurodegenerative disease. There has been a rapid increase in the knowledge of epidemiology, genetics, risk factors, and underlying neuropathological mechanisms, but still there is no cure for AD. Recent promising studies with functional imaging using positron emission tomography (PET) and magnetic resonance imaging reveal that disease processes can be detected when very early subjective symptoms of AD are manifest. Recently the PET ligand PIB was reported to bind in vivo to beta-amyloid in the brains of AD patients. Also cerebrospinal fluid markers including tau, phosphotau, and A beta 1-42 are probably important early biological markers that will provide an early diagnosis of AD. An obvious impairment in central cholinergic transmitter function and its close relation to cognitive function led to the development of the acetylcholinesterase inhibitors that now are used as symptomatic therapy. A drug interfering with the glutaminergic brain transmitter system, the NMDA antagonist memantine, has recently been approved for the treatment of patients with severe AD. In order to stop or reverse disease progression, different AD treatment strategies are of great interest. Epidemiological studies support the hypothesis that long-term treatment with estrogen, antioxidants, anti inflammatory drugs, and cholesterol-lowering agents could protect against the development of AD. Treatment with these drugs in manifest AD has been less promising. The use of nerve growth factors was limited by severe side effects. Much evidence supports the key role of beta-amyloid in the pathogenesis of AD. Compounds such as amyloid beta-sheet breakers, cholesterol-lowering drugs, estrogen, nicotine, zinc and copper chelators, inhibitors of beta- and gamma secretases, and immunization to reduce the amyloid burden in transgenic mice overexpressing beta-amyloid all have their advocates. The latter exciting strategy turned out to cause meningoencephalitis in 6% of AD patients so treated. One patient from the trial has died showing less beta-amyloid burden in brain than expected and patients with serum beta-amyloid plaque reactive antibodies had less cognitive decline after 1 year than AD patients without antibodies. There is a great optimism for early diagnosis and effective treatment of AD in the future. PMID- 14756160 TI - Depression in later-life Puerto Rican primary care patients: the role of illness, stress, social integration, and religiosity. AB - BACKGROUND: Older Puerto Ricans belong to two rapidly growing demographic groups known to have high rates of depression: the aging and Hispanic populations. Studies of depression in Puerto Ricans have primarily focused on the impact of demographic factors and health. This study expands previous research, examining the relationships between depression and social stressors, social support, and religiosity, for Puerto Rican primary care patients aged 50 and older. PATIENTS: Participants included 303 Puerto Ricans from six primary care clinics in a northeastern city. METHODS: Patients completed in-person interview in Spanish. The Composite International Diagnostic Interview indicated depressive disorders meeting DSM-IV criteria. Bivariate and multivariate relationships between depression and demographics, health, social stress and support, and religiosity were explored. RESULTS: One fifth of participants met DSM-IV criteria for major depression or dysthymia. Participants with the lowest income, more recent migration, and poor subjective health were significantly more likely to be depressed. In addition, rates of depression increased steeply for patients caring for grandchildren and those with personal or family legal problems. Seeing few relatives each month and needing more instrumental, emotional, or financial support were also related to higher rates of depression. Unexpectedly, low objective illness severity correlated with increased depression, whereas religiosity and religious participation had no relationship to depression. CONCLUSIONS: The findings presented here indicate the potential for social stressors and inadequate supports to substantially increase the risk of depression in older Puerto Ricans in primary care settings. Further studies should explore incorporating these social risk factors into improved prevention, clinical detection, and culturally sensitive treatment of older depressed Puerto Ricans. PMID- 14756161 TI - Reliability of the Geriatric Depression Scale for use among elderly Asian immigrants in the USA. AB - The increasing numbers of Asian and other immigrants in the United States have resulted in greater demands for research methodology sensitive to cross-cultural issues. A regional probability sample (n = 407) of Asian elderly immigrants of different nationalities (Chinese, Korean, Indian, Filipino, Vietnamese, or Japanese) residing in New York City was used to examine the reliability of the Geriatric Depression Scale (GDS). Using the 30-item GDS, about 40% of this representative sample of Asian elderly immigrants was considered to be depressed, indicating higher depression rates than in the previous studies of other Asian elderly samples in the US and in Asia. Results also showed that the 30-item GDS and 15-item GDS Short Forms were reliable measures to assess depression in community-dwelling Asian immigrant elders. Data strongly suggest that Asian elderly immigrants in the US are at risk of depression, indicating a need for the design of culturally sensitive mental health programs. PMID- 14756162 TI - Quantitative electroencephalography in late-onset schizophrenia. AB - BACKGROUND AND OBJECTIVES: A variety of electroencephalogram (EEG) abnormalities may occur in schizophrenics who had a typical onset of the disorder in early adulthood. The purpose of this study was to investigate EEG findings in patients with late-onset schizophrenia (onset of illness between ages 40 and 60). METHODS: Ten patients (one male, nine female, average age 52.8 years old) with late-onset schizophrenia underwent magnetic resonance imaging (MRI) of the brain and recording of quantitative EEG. RESULTS: MRI results for seven of the subjects were within normal limits and the other three showed nonspecific changes. Quantitative analysis of EEG variables revealed that for eight of the subjects, absolute power and relative power for the four EEG frequency bands, overall mean frequency, mean frequency for each of the EEG frequency bands, and interhemispheric coherence values did not vary significantly from a normative database of age-matched normal subjects. CONCLUSIONS: Results obtained to some degree replicate the single previous EEG study of late-onset schizophrenia. These findings suggest that the neurophysiological etiology and mechanism of late-onset schizophrenia may differ from that of schizophrenia with a typical age of onset, and that brain abnormalities are not an inevitable accompaniment to late onset of schizophrenia. PMID- 14756163 TI - Attitudes of patients with mild dementia and their carers towards disclosure of the diagnosis. AB - OBJECTIVE: To determine the attitudes of patients with mild dementia and their carers towards the disclosure of diagnoses of cancer and dementia and whether there are differences between these groups. To determine whether any major adverse events occurred following disclosure of diagnosis of dementia. DESIGN: A prospective study followed by a retrospective case-note study after 1 year. PARTICIPANTS AND SETTING: Fifty patients with mild dementia and their carers were recruited from a memory clinic in Nottingham, UK, and an old-age psychiatrist, using a semistructured questionnaire, separately interviewed them. MEASURES: Questions related to whether patients and their carers wished to be told diagnoses of cancer and dementia; the reasons for this; whether they would want treatment and make use of predictive testing if available were included. At 1 year follow-up whether antidepressants had been prescribed and whether any catastrophic reaction had occurred following disclosure of diagnosis. RESULTS: An overwhelming majority of patients with mild dementia wished to be informed of their diagnosis (92%); even more (98%) of the same patient sample reported wanting disclosure of a hypothetical diagnosis of cancer. A higher proportion (98%) of carers would wish to be told if they were to develop either dementia or cancer. All patients and carers would like access to treatment for dementia. A total of 88% of patients and 86% of carers would make use of a predictive test of Alzheimer's disease. Before receiving a diagnosis of mild dementia, only 28% of patients had insight that they may have dementia. Only a quarter (26%) of carers did not want the doctor to disclose the diagnosis of dementia to the patient. After 1 year, 6% of the original sample had developed a depressive illness requiring antidepressant treatment and no major incidents had occurred following disclosure of diagnosis. CONCLUSIONS: There has been a growing debate on whether patients with dementia should be informed of their diagnosis. Until recently, little was known about the views of patients themselves. This study shows that the vast majority of patients with mild dementia wish to be fully informed. Despite increasing awareness, a quarter of carers still do not wish their relative to be informed, though wish to be informed if they themselves were to develop the illness. This is significantly lower, however, than previously reported, suggesting a shift in attitudes. This study adds support to the weight of evidence that disclosure of a diagnosis of dementia does not cause depression or any irreversible harm to the patient. PMID- 14756164 TI - Hoarding behavior in the elderly: a comparison between community-dwelling persons and nursing home residents. AB - OBJECTIVE: To determine correlates of hoarding behavior in frail elderly persons. METHODS: Information about nursing home residents (n = 408) and community dwelling senior day-care participants (n = 177) was gathered through interviews with family and professional caregivers, medical chart review, and physician examinations, and included the following areas of assessment: hoarding behavior, demographic and health information, level of cognitive functioning, activities of daily living (ADL) performance, depressed affect, social functioning, manifestations of agitated behaviors, and previous stressful life experiences. RESULTS: We found that 15% of the nursing home residents and 25% of the community dwelling senior day-care participants manifested hoarding behavior at a rate of several times a week or higher. For nursing home residents, hoarding behavior was significantly related to a larger appetite, taking fewer medications, higher social functioning, comparatively less ADL impairment, and manifestations of physically nonaggressive agitated behaviors. For senior day-care participants, hoarding behavior was significantly associated with being female, a larger appetite, comparatively less gait impairment, fewer medical diagnoses, more involvement in activities, a positive diagnosis of dementia, hallucinations, the delusion of infidelity, and manifestations of three syndromes of agitated behaviors. CONCLUSION: While hoarding behavior in our samples presents differently from compulsive hoarding described in the literature, we obtained robust findings that show that despite differences in living conditions, the elderly persons who manifested hoarding behavior were those with relatively fewer health and functional disabilities. In addition, those who exhibited hoarding behavior also manifested agitated behaviors. We suggest that future researchers develop alternative measures of hoarding behavior so as to further clarify the phenomenon of hoarding behavior in the elderly. PMID- 14756165 TI - Treatment of aggressive behavior in dementia with the anticonvulsant topiramate: a retrospective pilot study. PMID- 14756166 TI - Introduction. Neutron and photon spectrometry techniques for radiation protection. PMID- 14756167 TI - Concepts and quantities in spectrometry and radiation protection. PMID- 14756168 TI - Neutron spectrometry in mixed fields: multisphere spectrometers. PMID- 14756169 TI - Neutron spectrometry in mixed fields: proportional counter spectrometers. AB - Proton recoil proportional counters have been successfully used in many laboratories worldwide for more than 30 years in order to measure high-resolution neutron energy spectra. The method is well elaborated and understood, nevertheless high expertise is required for its proper application. Table 4.1 summarises typical basic data for proton recoil proportional counters and the requirements for their application. It is noteworthy that any limiting parameter can vary to a certain degree depending on the quality of the detectors used (design, gas purity, response functions, etc) and their response matrices, the data analysis and the unfolding procedures applied with a correct evaluation of the uncertainties involved. It is also important to lend a critical eye to details during measurements (e.g. environmental and electronics problems) as well as in subsequent analysis and unfolding (e.g. oscillations due to unfolding artefacts or inadequacies in detector response matrices). It is recommended that any spectrometry system (procedures for measurement and data evaluation) should be tested and validated in well-known neutron fields e.g. 252Cf standard fission or 241Am-Be. One should, however, expect that, due to different room scatter conditions, deviations from the ISO spectra may occur, especially for low neutron energies. In order to demonstrate the capability of the recoil proton counter technique, two examples of typical neutron spectra are shown in Figures 4.20 and 4.21, both measured in mixed neutron-gamma fields at nuclear research reactors. PMID- 14756170 TI - Neutron spectrometry in mixed fields: NE213/BC501A liquid scintillation spectrometers. PMID- 14756172 TI - Photon spectrometry in mixed fields. PMID- 14756171 TI - Neutron spectrometry in mixed fields: superheated drop (bubble) detectors. AB - The BINS neutron threshold spectrometer permits the analysis of the main features of a neutron field for radiation protection purposes. The system offers a virtually complete photon discrimination and nested threshold responses to neutrons, which allow the use of very effective 'few-channel' unfolding procedures. To date, the practical operating energy range of a BINS is 0.1-10 MeV, over which a resolving power of 20-30% can be expected when the deconvolution is performed without explicit pre-information. Spectrum unfolding results in relatively high uncertainties on the differential fluence distributions, but due to negative correlations in adjacent energy groups the uncertainties on integral quantities such as dose equivalent are small and of the order of 5% to 10%, similar to the results of other active spectrometers. In comparison with most radiation detectors, the BINS is an extremely slow system due to the intrinsic duration of a bubble pulse and to the time associated with pulse analysis. For example, the maximum sustainable fluence rate of 1 MeV neutrons is about 10(4) cm(-2) s(-1), which is low for many neutron physics experiments. However, this rate corresponds to an ambient dose equivalent rate of about 1 mSv h(-1), making the active device adequate for radiation protection applications in the workplaces described in Section 1. There are ample margins for improvement of the spectrometer. In particular, in the low-energy region a thermal-epithermal neutron group may be added by using chlorine-bearing emulsions stabilised at suitable temperatures. In fact, the latest version of the system achieves this goal by using a single superheated emulsion of dichlorotetrafluoroethane (R-114) operated at temperatures up to 55 degrees C. This extends the range of the spectrometer and at the same time removes the undue enhancement of the UNFANA output in the low energy region. Above 10 MeV, the resolution can be improved by adding more thresholds, e.g. by starting from a lower initial temperature and using finer temperature increments. Based on neutron kinematics, the theoretical upper energy threshold which can be generated with superheated emulsions is greater than 100 MeV. However, this would most likely require refrigerating the detectors, while the current simpler approach is to operate the detectors at incremental temperature steps starting from the ambient temperature. A range that should be easily achieved in practice is from thermal energies to 20 MeV. PMID- 14756173 TI - Determination of direction and energy distributions. PMID- 14756174 TI - Unfolding procedures. PMID- 14756175 TI - Quality assurance. PMID- 14756176 TI - Applications. PMID- 14756177 TI - High-energy neutron depth-dose distribution experiment. AB - A unique set of high-energy neutron depth-dose benchmark experiments were performed at the Los Alamos Neutron Science Center/Weapons Neutron Research (LANSCE/WNR) complex. The experiments consisted of filtered neutron beams with energies up to 800 MeV impinging on a 30 x 30 x 30 cm3 liquid, tissue-equivalent phantom. The absorbed dose was measured in the phantom at various depths with tissue-equivalent ion chambers. This experiment is intended to serve as a benchmark experiment for the testing of high-energy radiation transport codes for the international radiation protection community. PMID- 14756178 TI - Experimental and computational determination of neutron dose equivalent around radiotherapy accelerators. AB - An experimental and computational investigation of the neutron dose equivalent in the treatment room and maze of various radiotherapy accelerators in Canada was completed. A bubble detector was selected for the bulk of the measurements due to its relative insensitivity to gamma radiation. The rooms and accelerator heads were modelled using the MCNP4B Monte Carlo radiation transport code, modified with a photoneutron patch. The results of the investigation showed generally good agreement between the experiments. Monte Carlo and analytical approximations to the neutron dose equivalent, and suggest that if the analytical approximations are used carefully they may be substituted for more costly experimental or Monte Carlo determinations. PMID- 14756179 TI - Effectiveness of customised neutron shielding in the maze of radiotherapy accelerators. AB - An investigation was performed to examine the neutron dose equivalent in a radiotherapy maze lined with a customised neutron shielding material. The accelerator investigated was a Varian Clinac 2100C/D using 18 MV photons, and the neutron shielding utilised at this centre was Premadex commercially available neutron shielding. Based on Monte Carlo simulations, properly installed customised neutron shielding may reduce the neutron dose equivalent by up to a factor of 8 outside the maze, depending upon the installation. In addition, it was determined that the neutron dose near the entrance to the maze may be reduced by approximately 40% by using customised neutron shielding in the maze, as compared with a facility not using this shielding. This would have a positive dose-saving effect in doorless maze designs. PMID- 14756180 TI - Development of a new passive integral dosemeter for gamma ray monitoring using an imaging plate. AB - A new passive integral dosemeter for gamma ray monitoring is being developed using an imaging plate (IP). In the application of IPs to dosimetry, a fading effect causes serious problems. The fading is considered to be caused by the recombination or holes and charges trapped shallowly in the F centres, which have several activation energies. Appropriate annealing procedures allow elimination of trapped charges with low activation energies, and quantitative estimation of the radiation dose is possible. The optimum condition for minimising the effect of fading on dose estimation was obtained by post-irradiation annealing of BAS-MS (BAS-MS2025, fabricated by Fuji Film Co. Ltd) IPs at 80 degrees C for 24 h. This was confirmed by calculation. Under this condition, the detection limit was calculated to be 3.15 microSv when 10% error is allowed. A 1-month integral dose was measured by the IP. The results were compared with those obtained using a commercially available fluoro-glass dosemeter. PMID- 14756181 TI - Dose measurement for patients and physicians using a glass dosemeter during endovascular treatment for brain disease. AB - It has been reported that exposure of patients and physicians to radiation from interventional radiological procedures cannot be disregarded. Direct measurement of patient exposure used to be difficult due to possible interference by the detector with the observation of X ray images. Recently, a dosemeter system consisting of small-sized glass chips and a reader which adopts pulsed UV laser stimulation has been developed. Owing to its small size, radiolucency and physical characteristics, direct monitoring of surface dose has become feasible. Dose measurement for patients and physicians during neurointervention was done using the photoluminescence glass dosemeter system. The dose-response of the dosemeter was almost linear over a broad dose range, but its energy dependency was rather high without a filter, the use of which is recommended by the manufacturer to compensate for energy dependency. Variation of sensitivity of about 20% was observed for effective energies of 45-60 keV which are used in neurointervention. In spite of this shortcoming, the photoluminescence glass dosemeter system was judged to be a convenient means for monitoring dose during neurointervention. PMID- 14756182 TI - Limitation of individual internal exposure by consideration of the confidence interval in routine personal dosimetry at the Chernobyl Sarcophagus. AB - In view of the probabilistic nature and very wide uncertainty of internal exposure assessment, its deterministic ('precise') assessment does not protect against not exceeding established reference levels or even the dose limits for a particular individual. Minimising such potential risks can be achieved by setting up a sufficiently wide confidence interval for an expected dose distribution instead of its average ('best' estimate) value, and by setting the limit at the 99% fractile level. The ratio of the 99% level and the mean ('best' estimate) is referred to as the safety coefficient. It is shown for the typical radiological conditions inside the Chernobyl Sarcophagus that the safety coefficient corresponding to the 99% fractile of the expected internal dose distribution varies within the range from 5 to 10. The maintenance of minimum uncertainty and sufficient sensitivity of the indirect dosimetry method requires measurement of individual daily urinary excretion of 239Pu at a level of at least 4 x 10(-5) Bq. For the purpose of reducing the uncertainty of individual internal dose assessment and making dosimetric methods workable. it is suggested that the results of workplace monitoring are combined with the results of periodic urinary and faecal bioassay measurements. PMID- 14756183 TI - Calibration and evaluation of a transportable in vivo monitoring system for accident monitoring of internal contamination. AB - A transportable in vivo monitoring system has been constructed and calibrated. The system uses two hyper pure germanium detectors--one for measuring whole body activities, by measuring activity in the torso, and the second for determining activities of radioiodine in the thyroid. The optimum counting geometries have been determined and the system has been calibrated for subjects of different ages and builds. The complete system is transported in two trailers which are pulled by ordinary motor vehicles. The minimum detectable activity (MDA) for 137Cs in whole body for a 10 min counting interval at the 95% confidence level is 200 Bq. The MDA for a count of 131I in thyroid is 20 Bq. The system is capable of detecting activities that are equivalent to a dose of 1 mSv for a wide range of radionuclides. PMID- 14756184 TI - Indoor radon levels and lung cancer risk estimates in seven cities of the Bahawalpur Division, Pakistan. AB - Indoor radon concentration levels were measured in seven major cities of the Bahawalpur Division, Pakistan. These included Fort Abbas, Minchin Abad, Hasilpur, Bahawalpur, Liaqatpur, Rahimyar Khan and Sadiq Abad. In order to select houses for this survey, the inhabitants were approached through their school-registered children. Due to several constraints, only those 100 houses were chosen in each city that were relatively the best representatives of the built-up area. The selected houses were then divided into live categories according to the house locations and building characteristics. CR-39 detectors, placed in polyethylene bags. were installed at head height in bedrooms and sitting rooms of all the selected houses and were exposed to radon and its daughter products for 90 days. Four such measurements were performed over a year in order to average out the seasonal variation in radon levels. After exposure, all the detectors were etched and counted under an optical microscope. The track densities of four measurements were averaged out and related to radon concentration levels. The radon levels were found to be 20, 20, 26, 28, 34, 42, 47 Bq m(-3) in the bedrooms and 24, 26, 27, 26, 37, 40, 43 Bq m(-3) in sitting rooms of Hasilpur, Rahimyar Khan, Minchin Abad, Fort Abbas, Sadiq Abad, Bahawalpur and Liaqatpur respectively. The observed variation in the radon level may be attributed to the geological variation in the area. Based on the observed data, excess lung cancer risk was assessed using the risk factors recommended by the USEPA, UNSCEAR and the ICRP. According to the EPA model, the lifetime excess lung cancer risk due to the lifetime exposure is found to vary from 12-102 per million per year in the houses surveyed. This variation is from 16-114 and 26-62 per million per year if UNSCEAR and ICRP limits are applied respectively. PMID- 14756185 TI - On the interpretation and use of neutron calibration coefficients. AB - Calibration laboratories provide measurement services that include determining the calibration coefficients for neutron survey meters and personal dosemeters. While there are numerous documents dealing with the procedures for calibration of neutron measuring devices, the purpose of this paper is to clarify the use of the particular dose equivalent conversion coefficients used at the National Institute of Standards and Technology and the Pacific Northwest National Laboratory for these calibrations. PMID- 14756186 TI - Batch-to-batch variation in the TL glow peaks and sensitivity in the production of CaSO4:Dy TLD phosphor. AB - The thermoluminescence (TL) glow peak height ratio of the main dosimetric peak (at about 240 degrees C) and the lower temperature satellite peak (at about 140 degrees C) of CaSO4:Dy was found to vary between 1.4 and 11.5 in more than 100 batches of CaSO4:Dy TLD tested in the last 5 years. Efforts were made to minimise the batch-to-batch variation. In most of the batches (90% of cases), the peak height ratio was more than 5 and the variation in the TL sensitivity (with respect to the reference batch) rarely exceeded 10%. The study of the grain size dependence of the peak height ratio of ground and unground and with and without acid wash of phosphor grains indicated that the surface effects contribute significantly to the batch-to-batch variation. Crystallisation of phosphor grains was found to be affected considerably by the interruptions in the process of evaporation of acid during the preparation. Phosphor grain size was found to be an important parameter for maintaining the quality of production of CaSO4:Dy TLD phosphor. PMID- 14756187 TI - Characterisation of an electronic radon gas personal dosemeter. AB - The monitoring of radon exposure at workplaces is of great importance. Up to now passive measurement systems have been used for the registration of radon gas. Recently an electronic radon gas personal dosemeter came onto the market as an active measurement system for the registration of radon exposure (DOSEman; Sarad GmbH, Dresden, Germany). In this personal monitor, the radon gas diffuses through a membrane into a measurement chamber. A silicon detector system records spectroscopically the alpha decays of the radon gas and of the short-lived progeny 218Po and 214Po gathered onto the detector by an electrical field. In this work the calibration was tested and a proficiency test of this equipment was made. The diffusion behaviour of the radon gas into the measurement chamber, susceptibility to thoron, efficiency, influence of humidity, accuracy and the detection limit were checked. PMID- 14756188 TI - Angle dependence of signal currents from cylindrical ionisation chambers. AB - A signal current from a cylindrical ionisation chamber with an ionisation volume of 62.7 cm3, 40 mm in diameter and 50 mm long, peaked when the chamber was lixed at 0 degrees and at 90 degrees in 137Cs and 60Co gamma ray fields for source chamber distances of 1 m and 2 m. A smaller ionisation chamber showed a small peak at 0 degrees in both fields but not at 90 degrees. However, calculations indicated that the signal current from the smaller chamber would also show a peak at 90 degrees in a 137Cs point-source gamma ray field. Peaks occur because gamma rays attenuate along the cylindrical side wall or along the end walls when a chamber is tilted slightly from 0 degrees or 90 degrees and the direction of the gamma ray beam agrees with the plane of one of these walls. These facts suggest the need for care in the common practice of measuring and calculating responses for cylindrical ionisation chambers fixed perpendicular to gamma ray beams. PMID- 14756189 TI - Effects of D-003, a mixture of long-chain aliphatic primary acids, fluvastatin and the combined therapy of D-003 plus fluvastatin on the lipid profile of normocholesterolemic rabbits. AB - D-003 is a mixture of long-chain aliphatic primary acids isolated from sugar cane wax with cholesterol-lowering effects proven in animals and healthy human volunteers. D-003 reduced serum total cholesterol (TC) and low-density lipoprotein cholesterol (LDL-C) in rabbits, while it increased high-density lipoprotein cholesterol (HDL-C) and did not affect triglycerides. D-003 inhibits cholesterol synthesis by regulating, instead of directly inhibiting, hydroxamethylglutaryl-CoA (HMGCoA) reductase activity. Although the ways in which D-003 and statins inhibit cholesterol biosynthesis are not identical, the strong competitive inhibition of cholesterol biosynthesis induced by statins suggests that an enhanced decrease of LDL-C and TC caused by the combined therapy D-003 plus statins is not expected. Nevertheless, taking into account the differential effects of D-003 and statins in HDL-C and triglycerides in rabbits, potential benefits of such combined therapy on other lipid variables cannot been discarded. Fluvastatin is a statin that inhibits competitively HMGCoA reductase, like other members of this class. This study was undertaken to compare the cholesterol lowering effects of D-003, fluvastatin and the combined therapy of D-003 plus fluvastatin in normocholesterolemic rabbits. Animals were randomly distributed into four groups of eight. One control group received the vehicle, two groups were treated with D-003 or fluvastatin at 5 mg/kg/day each, and the fourth group received the combined therapy of both drugs at 5 mg/kg/day each. Treatments were orally administered for 30 days. Body weight, food consumption and overall animal behavior were recorded to detect any warning sign resulting from combined therapy. After treatment, it was found that both D-003 and fluvastatin had significantly lowered LDL-C - D-003 by 81.5% (p < 0.01) and fluvastatin by 61.4% (p < 0.05). Combined therapy reduced LDL-C values (75.9%). Final values and percent changes reached in all groups were different from the control (p < 0.01). The reductions of TC were consistent with LDL-C decreases, so that D-003, fluvastatin and combined therapy significantly lowered TC by 48.4% (p < 0.01), 39.7% (p < 0.05) and 45.3%, respectively, values being different from those of the control (p < 0.01). The responses of LDL-C and TC to combined therapy were statistically similar, but less pronounced than those reached by D-003 alone. D 003 and combined therapy, but not fluvastatin alone, increased HDL-C (+21.5% and + 19.0%, respectively), these changes being significant versus the control (p < 0.05). In turn, fluvastatin and combined therapy, but not D-003 alone, lowered triglycerides (13.6% and 13.0%, respectively, p < 0.05 versus control). The effects of combined therapy on HDL-C were similar to those of D-003 alone, and the effects of combined therapy on triglycerides were similar to those of fluvastatin alone. The only advantage of combined therapy appears to be that it shows better effects on HDL-C than those of fluvastatin alone and better effects on triglycerides than D-003 alone. No significant changes in lipid profile were observed in the control group. All groups showed similar food consumption and body weight gain, health status being unaffected by the treatments. It is concluded that D-003 and fluvastatin at 5 mg/kg/day administered orally for 30 days to normocholesterolemic rabbits lowered LDL-C and TC, D-003 being more effective in increasing HDL-C and fluvastatin in lowering triglycerides. Combined therapy did not improve the response of LDL-C and TC with respect to monotherapies, but induced better responses of HDL-C and triglycerides than fluvastatin alone had on HDL-C or D-003 alone had on triglycerides. PMID- 14756190 TI - Effect of D-003, a mixture of high molecular weight primary acids from sugar cane wax, on paracetamol-induced liver damage in rats. AB - D-003 is a mixture of very high molecular weight aliphatic primary acids purified from sugar cane (Saccharum officinarum, L) wax, in which the most abundant component is octacosanoic acid. Experimental studies have shown that D-003 not only shows cholesterol-lowering and antiplatelet effects, but also offers strong protection against plasma lipoprotein oxidation. Previous studies demonstrated that D-003 protected against the histological changes characteristic of Cl4C induced hepatic injury in rats. The aim of the present study was to investigate the effects of D-003 in acute hepatotoxicity induced by paracetamol in rats. Male Sprague Dawley rats were randomly distributed in two experimental series of three experimental groups as follows: group 1--positive control rats (paracetamol treated); groups 2 and 3--rats with liver damage induced by paracetamol and treated with D-003 at 5 and 25 mg/kg, respectively, and which also received paracetamol to induce liver injury. In experimental series 1, animals received paracetamol orally (600 mg/kg). In series 2, paracetamol was administered through the intraperitoneal route (200 mg/kg). Eighteen hours after paracetamol dosing, rats were anesthetized with ether and livers were removed for histopathological studies. In the two experimental series, D-003 at 5 and 25 mg/kg significantly (p < 0.01) decreased the percentage of turgent cells and hepatocytes with necrosis and increased the percentage of normal hepatocytes with respect to positive controls in a dose-dependent manner. Necrotic areas and inflammatory infiltrates were observed in the liver of nine out of ten (90%) positive controls. In turn, D 003 dramatically reduced both necrotic areas and inflammatory infiltrate and was present in only one out of ten (10%) animals treated in the two experimental series. No histological alterations in liver sections of negative controls were found. D-003 protected against the histological changes characteristic of paracetamol-induced hepatic injury in rats, in which the process of lipid peroxidation plays a major role. The relationship between this protective action of D-003 in this experimental model and its antioxidant effects needs to be further investigated before definitive conclusions are drawn. PMID- 14756191 TI - Differences in hormonal and inflammatory parameters in male Lewis and Long Evans rats with adjuvant arthritis. AB - The purpose of the present study was (i) to compare secretory responses of prolactin and corticosterone to the acute stress of immobilization in male rats of the Lewis and Long Evans strains and (ii) to compare secretion of the two hormones in rats with fully developed adjuvant arthritis (AA) and their relationship with the intensity of the inflammatory reaction. A short immobilization of 5 min induced equal elevations of both hormones in both strains, but 20-min immobilization produced significantly stronger responses in Long Evans rats than in Lewis rats. AA inhibited prolactin secretion equally in both strains (from 11.6 +/- 1.3 ng/ml to 4.2 +/- 0.6 ng/ml in Lewis rats, p < 0.01, and from 3.7 +/- 0.6 to 2.12 +/- 0.1 ng/ml in Long Evans rats, p < 0.05), but caused a conspiciously larger elevation of corticosterone in the Long Evans than in the Lewis animals (11.5 +/- 1.2 microg/dl in Long Evans rats versus 5.1 +/- 0.5 microg/dl in Lewis rats, p < 0.01) while basal levels were similar. The larger corticosterone response in the Long Evans rats was associated with a stronger inflammatory reaction assessed by hind paw swelling (2.3 +/- 0.1 ml for Long Evans rats versus 1.8 +/- 0.08 ml for Lewis rats, p < 0.01) and plasma levels of nitric oxide (47.5 +/- 5.7 microM for Long Evans rats versus 28.7 +/- 2.5 microM for Lewis rats, p < 0.01) than in the Lewis males with lower corticosterone levels. In conclusion, there are significant, obviously genetically based, differences in the corticosterone responses to both immobilization and AA between the two strains, with the Long Evans rats reacting more strongly than the Lewis rats. The lack of the expected inverse relationship between corticosterone levels and the intensity of the inflammation indicates that the activity of corticosterone is not its primary determinant and that other important factors are involved. PMID- 14756192 TI - Microvascular assessment in Behcet disease: videocapillaroscopic study. AB - The aim of this study was to evaluate microvascular assessment in patients with Behcet disease (BD) by means of an intravital videocapillaroscopic study. Sixteen BD patients were compared with an equivalent group of healthy subjects matched for age and sex. Videocapillaroscopy (VCP) was performed in peripheral areas and in conjunctiva, and morphological and quantitative parameters were assessed. In both areas VCP showed several morphological alterations (microaneurysms, megacapillaries, desertification areas) detectable in a high percentage of patients; quantitatively we found significant changes of incisuring and sludging score, of capillary loop intermediate branch length (in peripheral areas) and of arteriole/venule diameter (in conjunctiva). Therefore, vessel involvement included both the number and the whole vessel structure and was seen both in peripheral and conjunctival areas when the two different vascular beds of micro- and paramicrocirculation were examined. We conclude that an important rearrangement of microcirculation is detectable in BD and that VCP may have diagnostic and prognostic value, providing qualitative and quantitative information able to define the systemic extension of vascular damage and the degree of vessel wall alteration. PMID- 14756193 TI - Proapoptotic activated T-cells in the blood of children with Down's syndrome: relationship with dietary antigens and intestinal alterations. AB - Immune defects, thyroid abnormalities, infections and coeliac disease are often associated with Down's syndrome (DS). However, the basis of the immune defects is still unclear in DS. In the present study, we show that peripheral CD4 T-cells were decreased in children with DS, while mean values of cytotoxic CD8 T-cells were comparable with those from healthy children. Circulating activated (CD3/HLA DR positive) T-cells were increased and a large proportion of purified T-cells from DS were also positive for APO-I/FAS (CD95) antigen. To further explore the functional status of circulating activated T-cells, enriched CD3 lymphocytes were cultured for 3 h and were tested for positivity to annexin-V (ANX-V) and propidium iodide. T-cells with the early apoptotic phenotype were increased in cell cultures from DS children. Plasma levels of inteleukin-6 (IL-6) were higher in DS children than in healthy children. The incidence of coeliac disease was also increased in this group of children. Most DS children showed increased levels of circulating IgG or IgA specific for gliadin, and their plasma IL-6 levels correlated with those of antigliadin IgG. The number of CD4 circulating cells was very low in DS children with coeliac disease, was low in those with serum antigliadin antibodies and was normal in DS without antigliadin antibodies. An overload of dietary antigens and impaired nutrient absorption secondary to altered functioning of the gastrointestinal mucosa might interfere with normal immune responses by inducing programmed cell death in CD4 T-cells. PMID- 14756194 TI - Financing results and value in behavioral health services. AB - Current changes require that behavioral health care leaders understand how public and private financing mechanisms interact and how, now more than ever, behavioral health care leadership must span multiple systems and financing streams. Understanding how financing mechanisms work, what they create, and what they cause is essential if we are to make the most of increasingly limited and increasingly complex resource streams in today's health care market. This article explores a different paradigm of what adds value to publicly funded behavioral health care systems, and provides the framework for the American College of Mental Health Administration's call to behavioral health care administrators to take a new approach to the considerations behind funding decisions and payment mechanisms. PMID- 14756195 TI - Mental and substance use disorders among Medicaid recipients: prevalence estimates from two national surveys. AB - This paper presents national estimates of behavioral disorders among Medicaid recipients. The 12-month prevalence of 14 disorders in the National Comorbidity Survey was 48% (43% mental, 14% substance use), and of 6 disorders in the National Household Survey on Drug Abuse was 27% (21% mental, 9% substance use). Total and specific disorder rates are from 50% to more than 100% higher in Medicaid than in the total population, and exceed most other insurance status groups. By promoting detection and treatment of these disorders in Medicaid, mental health administrators and policymakers may reduce barriers to education, employment, family stability, and departure from welfare. PMID- 14756196 TI - Do patients who mismanage their funds use more health services? AB - One rationale for establishing programs that help patients manage their funds is that such patients make extensive use of expensive inpatient services. We surveyed the money management habits of 406 inpatients and determined their use of Veterans Administration (VA) services and related costs over the subsequent year. In multivariate analyses, there was no statistically significant relationship between need for money management and service use or cost. The misspending of funds for drugs may precipitate hospitalization for some outpatients. However, in a sample of hospitalized patients, mismanagement of funds was not associated with longer length of stay or increased service use following discharge. PMID- 14756197 TI - Performance measures and their benchmarks for assessing organizational cultural competency in behavioral health care service delivery. AB - A project is described in which performance measures of cultural competency in behavioral health care were selected and benchmarked. Input from an Expert Panel representing the four major ethnic and racial groups in the U.S. and persons with extensive experience in implementing cultural competency in health care, along with survey data from 21 sites were used in the process. Measures and benchmarks are made specific to organizations that administrate care networks, and to service entities that deliver care. Measures were selected to parallel an implementation process, and benchmarks were set at "gold standard" levels. PMID- 14756198 TI - Mental health services for patients with multiple sclerosis residing in long-term skilled nursing facilities: problems and recommendations. PMID- 14756199 TI - Public institutions in many developing countries gain on-line access to the Administration and Policy in Mental Health journal. PMID- 14756200 TI - The qualitative and quantitative traditions within mental health administration. PMID- 14756201 TI - What is your diagnosis? Osteodystrophy secondary to hyperparathyroidism. PMID- 14756202 TI - Spontaneous antenatal resolution of canine hydrops fetalis diagnosed by ultrasound. AB - A previously unreported syndrome of transient mid-gestational hydrops fetalis identified by ultrasound was diagnosed in 16 litters of 16 different dogs between November 1999 and May 2002. During this study period, a total of 161 canine pregnancies were diagnosed by ultrasound. A 17th litter of eight fetuses developed similar ultrasonographic changes concurrently with maternal systemic mastocytosis and subsequently spontaneously aborted. No pups were born with clinical signs of hydrops fetalis. Fetal resorption in the affected litters was 7/95 (7.4 per cent) and 8/95 (8.4 per cent) aborted. Of the fetuses that survived to term, there were 7/88 (8 per cent) stillbirths. Neonatal mortality rate in the affected litters was 15 per cent (11/73) and the incidence of congenital abnormalities was 7/73 (9.6 per cent). Pugs were significantly (22.8 times) more likely to be affected than other breeds. PMID- 14756203 TI - Spinal arachnoid pseudocysts in 10 rottweilers. AB - Ten rottweilers presenting with spinal arachnoid pseudocysts were investigated. In six dogs, the lesions were localised dorsally at C2-C3; in three dogs, dorsally and ventrally at C5-C6; and, in one dog, dorsally and ventrally at C6 C7. Clinical signs were consistent with focal compression of the affected spinal cord segments. The animals showed ataxia of all four limbs, with truncal ataxia and marked hypermetria in cases of C2-C3 involvement, or ambulatory tetraparesis in cases of C5-C6 or C6-C7 involvement. Other than signs indicative of spina bifida in one dog, no abnormalities could be detected on plain radiographs. Myelography was used to define the localisation and extent of the pseudocysts. Additional information was obtained using magnetic resonance imaging in five dogs. Five dogs underwent a dorsal laminectomy; in three cases, the pseudocyst was treated by marsupialisation and, in two, by durectomy. PMID- 14756204 TI - Management of cor triatriatum dexter by balloon dilatation in three dogs. AB - Two dogs, one immature and one adult, were presented with a history of progressive ascites. In a third, immature dog, increasing exercise intolerance had been noted. Echocardiography demonstrated a partition in the right atrium (cor triatriatum dexter) and echocontrast studies documented normal flow from the cranial vena cava into the right atrium and ventricle. A saphenous vein contrast study demonstrated flow from the caudal vena cava into an accessory right atrial chamber (sinus venarum). The sinus venarum communicated with the true right atrium via a small defect in the atrial membrane in one dog, and additionally with the left atrium via a right-to-left shunting foramen ovale in the other dogs. All defects were visualised on angiographic studies by selective catheterisation of the caudal vena cava via the femoral vein. Balloon dilatation of the defect was then performed using a small followed by a larger balloon angioplasty catheter to enlarge the defect in the atrial membrane. Clinical signs improved within days and were sustained in the long-term in all cases. PMID- 14756205 TI - Immune-mediated haemolytic anaemia associated with a sarcoma in a flat-coated retriever. AB - A seven-year-old flat-coated retriever presented with a history of lethargy, dyspnoea and inappetence of several days' duration. Clinical examination revealed pale mucous membranes and tachypnoea, and haematology demonstrated marked autoagglutination. Thoracic radiographs revealed an increased opacity in the perihilar region. The owners declined further evaluation and the dog was treated symptomatically with immunosuppressive doses of prednisolone and azathioprine. The dog's demeanour improved, although it was eventually euthanased seven weeks later because of dysphagia and worsening dyspnoea. Postmortem examination revealed a widespread, poorly differentiated sarcoma involving the lungs, pericardium, thoracic lymph nodes and spleen. Immune-mediated haemolytic anaemia is a well recognised condition in dogs and is occasionally associated with neoplastic conditions. This is the first case report to describe immune-mediated haemolytic anaemia associated with a diffuse, poorly differentiated sarcoma. PMID- 14756206 TI - Rupture of the biceps brachii tendon sheath in two dogs. AB - Rupture of the biceps brachii tendon sheath was diagnosed in two dogs which were presented with chronic thoracic limb lameness. In each case, diagnosis was achieved by positive contrast arthrography, which revealed obvious leakage of contrast agent from the distal portion of the tendon sheath. Arthroscopy was performed in each affected shoulder joint and no other significant lesions were found. In one dog, concomitant bicipital tendinopathy was confirmed by histopathology. Both dogs were treated by bicipital tendon transposition, and tenodesis and both showed improvement in the degree of lameness following surgery. Tearing of the biceps brachii tendon sheath has not been reported previously but should be included in the differential diagnosis for shoulder lameness in the adult dog. PMID- 14756207 TI - Tracheal narrowing secondary to airway obstruction in two cats. AB - Tracheal narrowing is described in two cats. In both cases, inspiratory radiographs demonstrated tracheal narrowing just cranial to the thoracic inlet; no narrowing was seen on expiratory radiographs. Magnetic resonance imaging revealed suspected nasal tumours in both cases, but no abnormalities were identified in the region of the narrowed trachea. Nasal biopsy confirmed intranasal lymphoma in one cat and nasal adenocarcinoma in the other. The former cat was treated with chemotherapy. The owner of the latter cat declined further treatment. The tracheal narrowing disappeared after the initiation of chemotherapy in the cat with intranasal lymphoma and it is suggested that the narrowing might have been associated with the nasal tumour. A careful evaluation of the airway, especially cranial to the narrowing, is recommended in cases of tracheal narrowing in cats. PMID- 14756209 TI - Adverse reactions to microchips. PMID- 14756208 TI - Shedding light on vision and visual impairment. PMID- 14756210 TI - Open your eyes to ophthalmologists. PMID- 14756211 TI - Pasteurella multocida peritonitis: another risk of animal-assisted therapy. PMID- 14756212 TI - Volume 25: An important milestone despite continuing infection control challenges. PMID- 14756213 TI - Knowledge, attitudes, and practices of food service staff regarding food hygiene in Shiraz, Iran. AB - BACKGROUND: The practice of safety measures by the food service staff in hospitals is necessary for the prevention of food-borne outbreaks. Hospitalized patients are more vulnerable to potential hazards, and neglecting these principles can lead to increased morbidity and mortality. METHODS: We assessed the knowledge, attitudes, and practices of food service staff regarding food hygiene in government and private hospitals in Shiraz, Iran. Two questionnaires were designed, one for food service staff and the other for supervisors. Thirty one hospitals were approached, and the response rate was 99.5%. Four models were developed regarding knowledge, attitudes, and practices, and a multiple logistic regression analysis was performed. Comparison among the government and private hospitals was done. RESULTS: This study showed that personnel had little knowledge regarding the pathogens that cause food-borne diseases and the correct temperature for the storage of hot or cold ready-to-eat foods. Older personnel had better attitudes and practices. Females practiced safety measures less often than did males. Personnel working in hospitals with fewer than 300 beds also had better practices. Most of the personnel had positive attitudes, but disparity between attitude and practice was noted. CONCLUSION: There is a dire need for education and increased awareness among food service staff regarding safe food handling practices. PMID- 14756214 TI - Does the architecture of hospital facilities influence nosocomial infection rates? A systematic review. AB - OBJECTIVE: To review the evidence regarding the effects of interventions to improve hospital design and construction on the occurrence of nosocomial infections. METHODS: Systematic review of experimental and non-experimental, architectural intervention studies in intensive care units (ICUs), surgical departments, isolation units, and hospitals in general. The studies dated from 1975, and were in English, French, German, Italian, and Spanish. Regardless of format, the studies were identified through seven medical databases, reference lists, and expert consultation. RESULTS: One hundred seventy-eight scientific articles were identified; however, none of these described a meta-analysis, systematic review, or randomized, controlled trial. Most of the articles were categorized at the lowest level of evidence (expert judgment or consensus statements). Only 17 described completed concurrent or historical cohort studies matching the inclusion criteria (ICUs, 9; surgical departments, 4; isolation units, 2; hospitals in general, 2). The interventions generally included a move to other premises or renovation. However, in many studies, the staff-to-patient ratio was also improved. Some studies showed lower infection rates after intervention, but this finding cannot be generalized because of confounding and frequently small study populations. CONCLUSIONS: The lack of stringent evidence linking hospital design and construction with the prevention of nosocomial infection is partly attributable to the multifactorial nature of these infections, and some improvement will be seen if basic conditions such as the availability of sufficient space, isolation capacity, and facilities for handwashing are met. However, to our knowledge, other factors, especially the improper hand hygiene of medical staff, have greater impact. PMID- 14756215 TI - Preliminary results for a new final package test to assess the quality of sterile package systems. AB - OBJECTIVES: To develop a microbial test method to ascertain the passage of airborne bacteria through the medical device packaging system after sterilization, and to apply this test method to flexible packages under mechanical pressure changes. METHODS: Petri dishes filled with nutrient agar were integrated into the packaging unit prior to sterilization. We examined paper packaging consisting of 1 (single-paper packaging [P]), 2 (double-paper packaging [PP] and textile and paper double packaging [TP]), and 3 (double packaging with transport packaging [TPP]) layers. After sterilization, the test packages were pressed five times per minute for 1 or 3 hours by a mechanical device weighing 1 kg. This exposure took place in rooms with an average airborne bacterial count of 35 (room 1) or 440 (room 2) CFU/m3. The packaging was opened following culture at 37 degrees C for 48 hours to determine the number of colonies formed. RESULTS: The proportion of contaminated packages rose with the duration of mechanical stress and increased airborne bacteria concentration. Thus, mechanical pressure change for 3 hours resulted in the contamination of 60% (P), 15% (PP), 0% (TP), and 0% (TPP) of the packages in room 1, whereas 100% (P), 65% (PP), 73% (TP), and 0% (TPP) of the packages in room 2 were contaminated. CONCLUSIONS: This test method allows sterile packaging systems to be tested for contamination under practical conditions without extensive laboratory preparation. Contamination as a result of laboratory errors can be ruled out almost certainly. PMID- 14756216 TI - Injection practices in Romania: progress and challenges. AB - OBJECTIVE: To identify breaks in infection control practices that might put Romanians at risk for transmission of hepatitis B virus (HBV) from injections. METHODS: A standardized questionnaire was administered to a systematic sample of the 1,906 nurses in Valcea District, Romania, to collect information on their knowledge, attitudes, and practices regarding injection administration and universal precautions. RESULTS: Of the 180 nurses interviewed, 91% (95% confidence interval [CI95], 86% to 95%) reported having attended training for universal precautions; 58% (CI95, 49% to 67%) accurately reported that HBV remains infectious for at least 1 week in the environment; and 4% (CI95, 2% to 8%) knew that HBV is transmitted more efficiently than HIV through percutaneous exposures. No nurses reported reusing syringes or needles on different patients, but 4 (2%; CI, 1% to 6%) would reuse a syringe and 3 (2%; CI95, 0% to 5%) would reuse a needle on the same patient in an emergency. Fifty-three percent (CI95, 44% to 61%) of nurses reported having a dedicated area for the preparation of injectable medications separate from where blood-contaminated items were handled. Shortages of infection control supplies were common. CONCLUSIONS: Although nurses in Valcea do not report reusing injection equipment without sterilization, other unsafe practices occur that may facilitate HBV transmission through injections, including preparing injectable medications in areas potentially contaminated with blood. Inadequate knowledge of blood-borne pathogen transmission and shortages of infection control supplies may contribute to these unsafe practices. Addressing these deficits could improve injection safety in Romania. PMID- 14756217 TI - Infection control programs in Italian hospitals. AB - OBJECTIVE: To quantify the frequency and features of infection control programs implemented in Italian public hospitals. METHODS: In 2000, a questionnaire was mailed targeting all teaching and research hospitals and those with more than 300 beds, and a random sample of 50% of the district hospitals with fewer than 300 beds. RESULTS: The overall response rate was 80%. Fifty percent of the 428 respondent hospitals claimed to have an infection control committee, 43% an infection control physician (average, 1 infection control physician per 2,963 beds), and 33% an infection control nurse (average, 1 infection control nurse per 572 beds). Having an infection control committee, nurse, and physician occurred significantly more frequently in Northern and Central Italy, where the Regional Authority had implemented a regional infection control policy, and in larger hospitals. Thirty-nine percent of the hospitals claimed to have ongoing surveillance in place, mostly based on laboratory results. Eighty percent of the hospitals had defined at least one written protocol related to infection control policies, mostly for housekeeping, cleaning, disinfecting and sterilizing patient equipment, or standard precautions; on the contrary, policies aimed at preventing specific infections were less frequent. CONCLUSION: This national representative survey showed that the infrastructure for infection control is suboptimal when compared with the guidelines and surveys published in other countries. PMID- 14756218 TI - An approach to the study of potentially preventable nosocomial infections. AB - OBJECTIVE: To analyze a method that identifies potentially preventable nosocomial infections, as a tool to evaluate the performance of infection control programs through quantification of their potential for reducing nosocomial infections. METHODS: The database of the Study of the Prevalence of Nosocomial Infections in Spain (EPINE) was reanalyzed. The method was based on the use of false negatives of the classification table obtained from application of a fixed multiple logistic regression model, as an estimator of the number of potentially preventable nosocomial infections. RESULTS: The calculated number of patients with preventable infections was 7,493, which constituted 21.6% of the infected patients. Among hospital areas, intensive care had the lowest preventability rate (4.6%), whereas gynecology and obstetrics had the highest (40.6%). There was a significant inverse exposure-effect relationship between the proportion of preventable infections and the National Nosocomial Infections Surveillance (NNIS) System risk index. No correlation was observed between the prevalence of patients with nosocomial infection and the percentage of preventable infections. CONCLUSION: This analysis suggests that fewer nosocomial infections may be preventable in Spanish hospitals than previously assumed. PMID- 14756219 TI - Effect of education and performance feedback on rates of catheter-associated urinary tract infection in intensive care units in Argentina. AB - OBJECTIVE: To evaluate the effect of education and performance feedback regarding compliance with catheter care and handwashing on rates of catheter-associated urinary tract infection (UTI) in intensive care units (ICUs). SETTING: Two level III adult ICUs in a private healthcare facility in Argentina. PATIENTS: All adult patients admitted to the study units who had a urinary catheter in place for at least 24 hours. METHODS: A prospective, open trial in which rates of catheter associated UTI determined during a baseline period of active surveillance without education and performance feedback were compared with rates of catheter associated UTI after implementing education and performance feedback. RESULTS: There were 1,779 catheter-days during the baseline period and 5,568 catheter-days during the intervention period. Compliance regarding prevention of compression of the tubing by a leg improved (from 83% to 96%; relative risk [RR], 1.15; 95% confidence interval [CI95], 1.03 to 1.28; P = .01) and so did compliance with handwashing (from 23.1% to 65.2%; RR, 2.82; CI95, 2.49 to 3.20; P < .0001). Catheter-associated UTI rates decreased significantly from 21.3 to 12.39 per 1,000 catheter-days (RR, 0.58; CI%, 0.39 to 0.86; P = .006). CONCLUSION: Implementing education and performance feedback regarding catheter care measures and handwashing compliance was associated with a significant reduction in catheter-associated UTI rates. Similar programs may help reduce catheter associated UTI rates in other Latin American hospitals. PMID- 14756220 TI - Footwear exchange has no influence on the incidence of febrile neutropenia in patients undergoing chemotherapy for hematologic malignancies. AB - OBJECTIVE: To determine whether footwear exchange affects the incidence of febrile neutropenia among patients undergoing chemotherapy for hematologic malignancies. DESIGN: Open trial with historical comparison. SETTING: The 12-bed high-efficiency particulate air-filtered hematology unit at Osaka University Hospital, Suita, Japan. PATIENTS: Those with hematologic malignancies who underwent chemotherapy from January 1997 through January 2003. Footwear exchange was discontinued in January 2000. METHODS: The surveillance system was based on the National Nosocomial Infections Surveillance System of the Centers for Disease Control and Prevention. Rates of febrile neutropenia were calculated for neutropenic patient-days (ie, days with neutropenia < 500/microL). RESULTS: From January 1997 through December 1999 and from February 2000 through January 2003, 58 and 54 patients endured 237 and 184 neutropenic periods following chemotherapy, and their total neutropenic days were 3,123 and 2,503, respectively. They showed episodes of febrile neutropenia 89 and 68 times, respectively. Infection rates were 28.5 and 27.2 per 1,000 neutropenic patient days (P = .83), respectively. CONCLUSION: The incidence of febrile neutropenia was not affected by footwear exchange. In hematology units, changing shoes does not appear to affect the rate of infections during neutropenic periods. PMID- 14756221 TI - Serial nosocomial transmission of Plasmodium falciparum malaria from patient to nurse to patient. AB - BACKGROUND: Nosocomial transmission of malaria is a rare phenomenon in the United States. OBJECTIVE: To describe the probable transmission of Plasmodium falciparum malaria from a patient to a healthcare worker and then from the healthcare worker to another patient. DESIGN: Case series. SETTING: Two community hospitals in Massachusetts. INTERVENTION: Routine medical and supportive care. MEASUREMENTS: Clinical and laboratory evaluation. RESULTS: A nurse developed falciparum malaria after a needlestick injury from a patient with documented falciparum malaria. Three days prior to her diagnosis, she cared for another patient, who subsequently developed falciparum malaria. That patient's parasite isolate genetically matched the nurse's isolate by two independent DNA fingerprinting techniques. CONCLUSION: After extensive evaluation, we believe that a nurse who had acquired falciparum malaria via needlestick subsequently transmitted malaria to another patient via a break in standard precautions. The implications of this mechanism of transmission are discussed. PMID- 14756222 TI - Susceptibilities of Candida species to amphotericin B and fluconazole: the emergence of fluconazole resistance in Candida tropicalis. AB - OBJECTIVE: To determine the susceptibilities of Candida species isolated from Taiwan to amphotericin B and fluconazole. DESIGN: Prospective surveillance study. METHODS: Each hospital was asked to submit up to 10 C. albicans and 40 non albicans Candida species during the collection period, from April 15 to June 15, 1999. One isolate was accepted from each episode of infection. The broth microdilution method was used to determine susceptibilities to amphotericin B and fluconazole. RESULTS: Only 3 of 632 isolates, one each of C. famata, C. krusei, and C. tropicalis, were resistant to amphotericin B. A total of 53 (8.4%) of 632 clinical yeast isolates, consisting of 4% C. albicans, 8% C. glabrata, 15% C. tropicalis, and 70% C. krusei, were resistant to fluconazole. In contrast, no C. parapsilosis isolate was resistant to fluconazole. Isolates from tertiary-care medical centers had higher rates of resistance to fluconazole than did those from regional and local hospitals (11.4% vs 6.6%). Isolates from different sources showed different levels of susceptibility to fluconazole. All of the isolates with the exception of C. tropicalis and C. krusei isolated from blood were susceptible to fluconazole. A pattern of co-resistance to both amphotericin B and fluconazole was observed. CONCLUSIONS: Non-albicans Candida species had higher rates of resistance to fluconazole than did C. albicans (44 of 395 [11.2%] vs 9 of 237 [3.8%]; P = .002). The increasing rate of fluconazole resistance in C. tropicalis (15%) is important because C. tropicalis is one of the most commonly isolated non-albicans Candida species. PMID- 14756224 TI - Performance improvement in the long-term-care setting: building on the foundation of infection control. AB - Infection control programs were among the first organized efforts to improve the quality of healthcare delivered to patients and are an excellent model for the development of other healthcare performance improvement activities. Whether labeled as infection control, quality improvement, or patient safety, performance improvement initiatives share similar methods and principles. The quality of care in long-term-care facilities (LTCFs) has been scrutinized for years and has received renewed attention with the recent initiation of public reporting of quality measures by Medicare. This article reviews the principles of performance improvement, discusses the importance of employing evidence-based interventions, and emphasizes the value of local performance improvement in LTCFs. Residents of LTCFs remain at high risk for the development of nosocomial infections, and among performance improvement initiatives, infection control is recommended as a high priority for all LTCFs. Fortunately, infection control contains the essential elements for performance improvement, and a successful infection control program can provide the foundation for expanding performance improvement throughout the LTCF. There is still much that needs to be done to determine the best clinical practices for LTCFs, and this should remain a priority for future research. Furthermore, efforts should continue to apply these principles at the local level to ensure that all residents of LTCFs receive the best care possible. PMID- 14756223 TI - Genetic analysis of Pseudomonas aeruginosa by enterobacterial repetitive intergenic consensus polymerase chain reaction (PCR) and arbitrarily primed PCR: gel analysis compared with microchip gel electrophoresis. AB - OBJECTIVES: To assess the applicability of a newly emerging microchip gel electrophoresis for rapid strain differentiation among clinical isolates of Pseudomonas aeruginosa, and to compare this technique with the traditional gel method for DNA separation. METHODS: One hundred clinical strains of P. aeruginosa obtained from a hospital in northwestern Ohio were tested for reactivity to 3 serotype-specific monoclonal antibodies by enzyme-linked immunosorbent assay. Twelve strains (4 from each serogroup) were selected for DNA analysis by polymerase chain reaction (PCR)-based, single primer DNA fingerprinting methods with 3 different primers: 1 enterobacterial repetitive intergenic consensus PCR and 2 arbitrarily primed PCRs. The PCR products were analyzed by agarose slab gel and microchip gel electrophoresis. RESULTS: Of the 100 clinical isolates tested, 39% (4%, 14%, and 21%) were found to be serotypes 0:3, 0:6, and 0:11, respectively. Twelve strains were chosen for DNA analysis by PCR. The PCR products were analyzed by agarose slab gel electrophoresis and on microchips to determine interspecies diversity. Both methods demonstrated that different serotypes exhibited different electrophoretic patterns. Two strains (clinical strains 6 and 7, serotype 0:6) showed identical patterns, indicating a high degree of relatedness. CONCLUSION: In all cases, there was concordance between the electrophoretic patterns detected by the two methods. The capability of conducting both PCR and microchip gel electrophoresis offers an opportunity for an automated and rapid method for genetic analysis and differentiation among strains of P. aeruginosa and other microorganisms. PMID- 14756225 TI - Independent prognostic factors for fatality in patients with urinary tract infection caused by Serratia marcescens. AB - In a retrospective study of 329 cases of nosocomial urinary tract infection caused by Serratia marcescens, 16 (4.9%) were fatal. Female gender (OR, 3.9; CI95, 1.3-11.7; P = .014) and secondary S. marcescens bacteremia (OR, 6.5; CI95, 1.5-28.6; P = .013) were independent prognostic factors for fatality. PMID- 14756226 TI - Clinical and microbiological profile of urinary tract infection at a tertiary care center in Lebanon. AB - We compared community-acquired urinary tract infection (UTI) with hospital acquired UTI at the American University of Beirut Medical Center. Escherichia coli was the most frequently isolated organism. Hospital-acquired E. coli isolates were often multidrug resistant. These results can be used to improve empiric treatment of UTI. PMID- 14756227 TI - Multicenter study of the prevalence of nosocomial infections in Italian hospitals. AB - A point-prevalence study of nosocomial infections was conducted in 10 general hospitals in northwestern Italy in June and July 2000. Infection rates were compared by type and site among the different hospitals. Urinary tract infections were most frequent, accounting for 57.8% of 128 nosocomial infections. PMID- 14756228 TI - Topical oxygen is not hyperbaric oxygen (HBO2). PMID- 14756229 TI - Productivity and acuity tool for wound care and hyperbaric medicine centers. AB - The unique workload and patient acuity in wound care and hyperbaric centers leaves staffing to subjective interpretation; lack of objective data makes it difficult to determine department budgets and appropriate staffing patterns. To address these problems, an objective tool was developed to assess acuity and productivity in hyperbaric and wound care departments. Patient classifications were created to define acuity based on disease and associated procedures. A total of 38 distinct indicators were identified for wound care and 22 for hyperbaric oxygen therapy. Time studies were performed on identified tasks. All patients were scored and placed in acuity classes. The number of hours per patient day was calculated and multiplied by direct care hours to find number of full-time equivalents (FTEs). The tool was tested for content validity and clinical applicability. The tool was found to accurately reflect patient-care work and suitable as a benchmarking tool for use in monoplace hyperbaric and wound care centers. PMID- 14756230 TI - An outbreak of Methicillin-resistant Staphylococcus aureus cutaneous infection in a saturation diving facility. AB - We present a molecular epidemiological investigation of an outbreak of cutaneous tissue infection, which involved six divers during a 45 day saturation exposure dive. The cutaneous infection manifested as boils, foliculitis and small abscesses involving different body sites, including nose, external ear canal, necks, back, extremities, and buttocks. Staphylococcus aureus was consistently isolated from the skin lesions of affected divers. A study of the antibiogram revealed that all Staphylococcus aureus isolates were uniformly resistant to penicillin, oxacillin and erythromycin, but sensitive to clindamycin, tetracycline, trimethoprim-sulfamethoxazole, rifampin and vancomycin. Molecular typing by pulse field gel electrophoresis (PFGE) demonstrated that all the Methicillin-resistant Staphylococcus aureus (MRSA) isolates had an indistinguishable pulsed field gel electrophoresis pattern. The source of outbreak was identified as a colonized diver (diver D). Personal contact was most likely the mode of transmission among the six divers. Infection with MRSA should be suspected in outbreaks of boils that are not responding to standard antibiotic therapy among healthy divers and their close contacts. To our knowledge, this is the first report of Methicillin-resistant Staphylococcus aureus (MRSA) outbreak in a saturation diving facility. PMID- 14756231 TI - Measurement of fatigue following 18 msw dry chamber dives breathing air or enriched air nitrox. AB - Many divers report less fatigue following diving breathing oxygen rich N2-O2 mixtures compared with breathing air. In this double blinded, randomized controlled study 11 divers breathed either air or Enriched Air Nitrox 36% (oxygen 36%, nitrogen 64%) during an 18 msw (281 kPa(a)) dry chamber dive for a bottom time of 40 minutes. Two periods of exercise were performed during the dive. Divers were assessed before and after each dive using the Multidimensional Fatigue Inventory-20, a visual analogue scale, Digit Span Tests, Stroop Tests, and Divers Health Survey (DHS). Diving to 18m produced no measurable difference in fatigue, attention levels, ability to concentrate or DHS scores, following dives using either breathing gas. PMID- 14756233 TI - Lack of toxic side effects in neutrophils following hyperbaric oxygen. AB - Conflicting data have been reported about the impact of repeated HBO2 exposure on the production of superoxide radicals during the neutrophil respiratory burst (RB) and on phagocytosis. In this study we wanted to see if exposure to hyperoxia would affect human neutrophil RB and phagocytosis. Short- and long-term effects after single or repetitive HBO2 exposure of 2.5 atmospheres absolute over a period of 90 min were studied in 40 healthy volunteers. The RB was measured by the intracellular oxidation of dihydrorhodamine after induction by Escherichia coli (E. coli), or priming with recombinant tumour necrosis factor alpha (TNF alpha), followed by N-formyl-methionyl-leucyl-phenylalanine (fMLP) stimulation. The phagocytic activity was determined by the intake of FITC-labelled opsonized E. coli. No differences could be found between RB and phagocytic activity before and after HBO2 therapy, regardless of short- or long-term exposure. These findings indicate that exposure to hyperoxia does not impair these two important functions of the human innate host defense. PMID- 14756232 TI - Change in strategy of solving psychological tests: evidence of nitrogen narcosis in shallow air-diving. AB - The depths from 10 to 30 m are usually not considered narcotic in scuba air diving, and evidence of psychomotor disturbances attributable to nitrogen narcosis at these depths is weak and contradictory. 15 experienced male divers were tested in a chamber at 1, 2, 3, and 4 bars over five consecutive days using a battery of computer generated psychological tests-Computerized Reactionmeter Drenovac (CRD-series). Total test solving time, minimal single task solving time, total "ballast" time, and total number of errors were recorded. Nitrogen narcosis effects were evident at all hyperbaric pressures with marked performance differences among subjects. MANOVA revealed significant effects of nitrogen partial pressure for groups of the same variables as follows: total test solving time (p < 0.001), total "ballast" time (p < 0.001), and total number of errors (p = 0.038), but not for minimal single task solving time. ANOVA showed significant effects of pressure only on tests of visual discrimination of signal location (total test solving time: p = 0.012, total "ballast" time: p < 0.001), simple convergent visual orientation (total test solving time: p = 0.012), and convergent thinking (total test solving time: p = 0.002, total number of errors: p = 0.049). The order of the pressure exposures had no influence on subject performance. Impaired psychomotor processing found during air exposures from 2 to 4 bars suggests that nitrogen narcosis at depths usually considered safe from its effects might be a problem in underwater operations that require accuracy, speed, limited time of performance, and complex psychomotor skills. PMID- 14756234 TI - Use of speech production repair strategies to improve diver communication. AB - The purpose of this investigation was to determine if speech intelligibility improved when divers made specific modifications to their speaking patterns while in a hyperbaric helium-oxygen (heliox) environment. Divers were trained to produce a variety of sentences using speech with three types of alterations: (1) slowed rate, (2) increased loudness, and (3) a combination of slightly slowed rate, a minimal increase in loudness, increased pause time, and greater mouth opening (composite strategy). Both diver and non-diver listeners judged these sentences for intelligibility. In addition, acoustic analysis of the cues for the identification of voicing, place, and manner of articulation was conducted to determine if such cues might become more audible in the speech signal when repair strategies were used. Both perceptual and acoustic results showed the composite method to be the best for natural-sounding, intelligible speech. It had the effect of slowing rate and increasing loudness just enough to increase intelligibility without causing distortion. It was concluded that teaching divers to produce speech using this method would be a worthwhile investment for improving speech intelligibility. PMID- 14756235 TI - Effect of hyperbaric oxygen on anastomoses created under the influence of 5-FU. AB - AIM: This study investigates the effects of hyperbaric oxygen (HBO2) therapy on the healing capacity of colonic anastomoses under the influence of preoperative chemotherapy. MATERIAL AND METHOD: Forty male Wistar-Albino rats were divided into four groups of 10. Colonic resection and anastomosis were performed in each group. Group I (control) received no further treatment. In group II, 5 fluorouracil was administered intraperitoneally for five consecutive days preoperatively. Group III received HBO2 therapy for seven days after the anastomosis. Group IV received HBO2 therapy following the administration of preoperative 5-fluorouracil. On the seventh postoperative day, all the rats were examined to determine the bursting pressures of the anastomosis and to take tissue sample from the anastomotic line for hydroxyproline measurement. RESULTS: Bursting pressures of the anastomosis in group IV were increased significantly compared to group II. Hydroxyproline levels were significantly increased with the use of HBO2 in rats, independent of chemotherapy administration. CONCLUSION: HBO2 therapy strengthens anastomoses created under the influence of neoadjuvant chemotherapy. This technique might have a future role in the care of colon cancer patients undergoing new multimodality cancer treatments. PMID- 14756236 TI - Evidence-based approach to HBO2 therapy. PMID- 14756237 TI - Osteoclast differentiation at growth plate cartilage-trabecular bone junction in newborn rat femur. AB - Using 3-day-old newborn rats, we examined the differentiation processes of osteoclasts associated with the destruction of the femoral growth plate cartilage and primary trabecular bone. In the growth plate cartilage, thin mineralized areas were detected solely in the longitudinal septal cartilage matrix in the hypertrophic zone, but the transverse septal cartilage matrix between adjacent chondrocytic lacunae within a row of chondrocytes remained unmineralized. The longitudinal septal cartilage between adjacent rows of chondrocytes appeared to persist, forming the walls of opened lacunar canals. Consistent with the removal of the transverse septal cartilage matrix, the longitudinal canals of opened chondrocytic lacunae were deeply invaded by capillary vessels, mononuclear cells and multinucleated pre-osteoclasts lacking a ruffled border. CD34-positive endothelial cells of capillary vessels deeply penetrated into the transverse septal cartilage matrix facing the medullary cavity and the opened chondrocytic lacunae. ED1-positive monocytes/macrophages were distributed at the chondro osseous junction, but they were distant from the erosive front of the transverse septa. Tartrate-resistant acid phosphatase-positive multinucleated pre osteoclasts lacking a ruffled border and differentiated osteoclasts with a ruffled border were localized mainly at two locations: the chondro-osseous junction and the growth front of primary bone trabeculae. Osteoclasts were located on the type-I collagen-positive bone trabeculae close to the growth plate, but they appeared to be distant from the type-II collagen-positive cartilage matrix. Even within opened chondrocytic lacunae, when osteoclasts were distant from the cartilage and bone matrix, they lacked polarized cytoplasmic organization and a ruffled border. The osteoclasts located in the remaining septal cartilage also exhibited neither a ruffled border nor a clear zone. Osteoclasts with a prominent ruffled border and clear zone were located in bone matrix covering the remaining septal cartilage. These results suggest that osteoclasts require hydroxyapatite crystals and bone matrix constituents for ruffled border formation and are not involved in resorption of the unmineralized transverse and mineralized longitudinal septal cartilage without covering bone matrix at the chondro-osseous junction. PMID- 14756238 TI - Defective bone remodelling in osteoprotegerin-deficient mice. AB - Previous studies have reported enhanced osteoclastogenesis, increased bone resorption and osteoporosis in osteoprotegerin (OPG)-deficient mice. In the present study, we show that the tibial epiphyses contain abundant, thin trabeculae lined with numerous osteoclasts and cuboidal osteoblasts. The increase in osteoblasts and osteoclasts was associated with a dramatic increase in calcein labelling of the mineralization fronts and replacement of much of the intertrabecular marrow with numerous alkaline phosphatase-positive preosteoblasts. Furthermore, the discrete, linear cement lines seen in wild-type mice were replaced by a randomly oriented meshwork of cement lines that were stained intensely for tartrate-resistant acid phosphatase and osteopontin in the OPG-/- mice. These indices of accelerated bone remodelling in mutant bone were associated with irregular trabecular surfaces, a disorganized collagen matrix interspersed with amorphous ground substance and numerous fissures between old and new bone. In total, these observations indicate that enhanced osteoclastic activity in OPG-/- epiphyses led to a coupled increase in osteoblast differentiation and activity and an increase in bone remodelling. The high bone turnover, disorganized matrix and impaired attachment of new to old bone in the cement lines in OPG-/- mice appear to cause bone fragility. PMID- 14756239 TI - Osteoclast differentiation and characteristic trabecular bone formation during growth plate destruction in osteoprotegerin-deficient mice. AB - Osteoprotegerin (OPG) is an osteoblast-derived secreted member of the tumour necrosis factor receptor superfamily that inhibits osteoclastogenesis. Mice that are OPG-deficient have severe bone loss, including growth plate cartilage destruction. Using OPG-deficient mice as a useful animal model, we attempted to clarify differentiation and ultrastructural features of osteoclasts located on destructed growth plate cartilage and trabecular bone matrix. In the humerus and femur of OPG homozygous (-/-) mice, adjacent to the growth plate cartilage, bone trabeculae without a calcified cartilage core were characteristically formed at the metaphyseal side of the medullary cavity, which resulted in an irregular chondrocyte distribution and arrangement in growth plate cartilage. During growth plate cartilage destruction, osteoclasts positive for tartrate-resistant acid phosphatase showed unusual localization on both type-II collagen-positive cartilage and type-I collagen-positive trabecular bone matrix at the ossification centre of the epiphyseal/metaphyseal border. Although multinucleated osteoclasts were distributed within open lacunar canals in the growth plate, those on uncalcified cartilage matrix lacked a ruffled border. Facing the calcified cartilage matrix within lacunar canals, osteoclasts showed irregularly formed ruffled borders. After growth plate destruction, a thin bone layer was deposited on the remaining cartilage surfaces by invading osteoblasts. Osteoclasts formed prominent ruffled border structures on bone matrix, deposited on the remaining growth plate cartilage. These results suggest that, in OPG (-/-) mice, terminal osteoclast differentiation requires the presence of newly produced bone matrix, as the coupled phenomenon of bone formation and resorption, as well as osteoblast derived cytokines. PMID- 14756240 TI - Regulatory mechanism of osteoclast activation. AB - Osteoclasts are multinucleated, terminally differentiated cells which play an essential role in bone resorption. Osteoclasts exhibit high expression of the alpha(v)beta3 integrin, which binds to a variety of extracellular matrix proteins, including vitronectin, osteopontin and bone sialoprotein. RGD (Aug-Gly Asp)-containing peptides, RGD-mimetics and blocking antibodies to alpha(v)beta3 integrin were shown to inhibit bone resorption in vitro and in vivo, suggesting that this integrin plays an important role in regulating osteoclast function. A number of signalling molecules were found to be involved in the alpha(v)beta3 integrin-dependent signalling pathway, including c-Src, Pyk2 and p130Cas. Both Pyk2 and p130Cas localize to the sealing zone of actively resorbing osteoclasts, suggesting their role in linking the adhesion of osteoclasts to the bone matrix, to cytoskeletal organization, and to the polarization and activation of these cells for bone resorption. In this article, we review the regulatory mechanism of osteoclast activation. PMID- 14756241 TI - Clathrin sheets on the protoplasmic surface of ventral membranes of osteoclasts in culture. AB - Physical cell-shearing resulted in various degrees of disruption of the basolateral (upper) membranes, cytoskeletons or cell organelles and exposed the protoplasmic surface of ventral (adhesion) membranes of osteoclasts that were attached to the underlying substratum, such as coverslips, mica or synthetic apatite plates. Freeze-dried replicas of the ventral membranes left behind on the substratum after cell-shearing provided three-dimensional information on the ultrastructure of the protoplasmic membrane surface of cultured osteoclasts. An extensive area of the protoplasmic surface and various amounts of cytoskeletal structures attached to the adherent ventral surface of the plasma membrane were visible. In particular, the most characteristic finding of the present study is that numerous clathrin sheets displaying various sizes, shapes and curvature were revealed on the ventral membrane. The polygon substructures of the clathrin lattices appeared to be composed of hexagons with a few pentagons interspersed. They were seen at the peripheral membranes where they were situated at the sites of close contact with the underlying substratum. In addition, clathrin lattices were never observed on the basolateral (upper) membranes. In favourable stereo views, most cytoskeletons were not in direct contact with the clathrin sheets. However, a few observations indicated possible remnants of cytoskeletons attached to clathrin lattices. Podosomes did not have a direct structural relationship to clathrin lattices. Although it is generally accepted that cytoskeletal podosomes in motile cells, such as osteoclasts, play a major role in cell adhesion, the present study indicates that membrane-associated clathrin might also function during attachment to the substrate. In this regard, clathrin is thought to be required for receptor-mediated endocytosis, but whether it might also function in cell attachment is still a matter for debate. This type of clathrin-related adhesion appears to be a previously unrecognized site of cell/substrate adhesion in osteoclasts. To assess this possible function, we focused on clathrin and related cytoskeletal elements on the ventral membranes of cultured osteoclasts. PMID- 14756242 TI - Extremely high expression of beta-actin mRNA in osteoclasts resorbing alveolar bone located at the distal area of the developing molar tooth germ in newborn rats. AB - Expression of beta-actin is widely utilized as an internal control of mRNA expression in various cells. Here we show evidence that the expression level of beta-actin mRNA in osteoclasts significantly differs in its intensity according to the position in bone tissues. By use of in situ hybridization, we obtained clear data showing that osteoclasts facing the distal part of a developing molar tooth germ expressed extremely high levels of beta-actin mRNA in comparison with other osteoclasts observed in the mandibular bone surface. No signal was detected when beta-nerve growth factor transcripts were observed. Electron microscopic analysis revealed that osteoclasts localized in these areas were functionally active, estimated from the expression of high levels of the osteoclast membrane antigen, Kat1 antigen, which associated with active osteoclasts. These data suggest that osteoclasts expressing extremely high levels of beta-actin are highly related to active osteoclasts induced by the mechanical stress caused by physiological movement of molar tooth germ towards distal directions in the developing mandible. PMID- 14756243 TI - Cathepsins in the osteoclast. AB - The mechanism by which bone collagen and other organic components are degraded by the osteoclast during osteoclastic bone resorption was unclear until the 1980s. Studies conducted since the early 1990s have identified lysosomal proteases, mainly cathepsins that are active at low pH, involved in osteoclastic bone resorption. Several cathepsins, such as cathepsins C, D, B, E, G and L, were initially demonstrated to take part in the degradation of organic bone matrix in osteoclasts. Cathepsin K, which has high proteolytic activity and localizes primarily in osteoclasts, was discovered in 1995. This first tissue-specific cathepsin was associated with pycnodysostosis, a genetic disorder observable as an osteopetrotic phenotype in cathepsin K-deficient mice. Cystatin C, an endogenous inhibitor of cysteine proteases, regulates the activity of cathepsin K. However, detailed morphological observations suggest that the organic bone matrix is degraded by not only cathepsin K, but also by matrix metalloproteinases or other cathepsins. The osteoclast possesses a unique endocytotic/exocytotic structure and each cathepsin is specifically localized in the osteoclast, which implies that each cathepsin contributes cooperatively to the process of osteoclastic bone resorption. Further studies may clarify the regulation of cathepsin activities and the roles of cathepsins during bone remodelling. PMID- 14756244 TI - Analysis of tooth formation by reaggregated dental mesenchyme from mouse embryo. AB - Tooth morphogenesis is a well-known developmental system related to epithelial mesenchymal interactions. In mice, the dental epithelium has the potential to induce tooth formation prior to the bud stage, whereas this potential shifts to the dental mesenchyme from the dental epithelium. The reaggregation of mesenchymal tissue leads to previous memories of individual cells being reset, which is useful for studying the predetermination of mesenchyme. Here, the mesenchyme was triturated into single cells after separation of the epithelium and the mesenchyme. These single cells were repelleted and combined with the epithelium. The reaggregated tooth was transplanted into a mice kidney capsule. In order to investigate the essential functions of both the dental epithelium and the dental mesenchyme regarding their mutual interaction, a reaggregation system was introduced using the late bud stage of the mouse first molar. Amelogenin expression was examined to confirm the cytodifferentiation in the reaggregated tooth. The results showed that a new tooth formed after reaggregating the dental mesenchyme. This tooth contained enamel, dentin, dentinal tubules and dental pulp. The inner enamel epithelium of the reaggregated tooth differentiated into ameloblasts. Immunohistochemistry for amelogenin was observed both in the ameloblasts and the enamel. However, the structure of the enamel was different from that of the normal tooth, with the thickness of the predentin becoming wider. These findings suggest that reaggregated dental mesenchyme cells can produce a tooth. The fate of dental epithelium was not affected by reaggregated dental mesenchyme, although the dental mesenchyme appears to lose the information from the dental epithelium. PMID- 14756245 TI - Lineage of non-cranial neural crest cell in the dental mesenchyme: using a lacZ reporter gene during early tooth development. AB - The tooth is one of the ectodermal organs controlled by reciprocal interactions between the epithelium and the mesenchyme. Mesenchymal cells in the developing tooth, so-called dental mesenchymal cells, are derived from two different origins: the cranial neural crest (CNC) and the non-CNC. These CNC-derived cells migrate, proliferate and differentiate into odontoblasts, cementoblasts, fibroblasts, osteoblasts and chondroblasts. Tooth germs of wild-type mice were transplanted into the kidney of adult lacZ-transgenic mice. After 1 week of transplantation, a few lacZ-expressing cells and many red blood cells were found near or inside the blood vessels in the pulp of wild-type tooth germs. This result shows that circulating cells of the adult host could invade the dental pulp during tooth development, through the blood vessels, and be a part of dental pulp tissue. Therefore, it can be suggested that these circulating progenitor cells could be the origin of non-CNC-derived cells in tooth germ and their migration pathways would be the blood vessels invading the dental pulp during tooth development. If variations of this experiment were suitably adjusted, such as the embryonic stage of the tooth germ, duration of transplantation, etc., this transplantation experiment using adult lacZ-transgenic mice could be a good system to reveal the origin and migration pathway of cells in developing organs as well as in dental mesenchymal cells. PMID- 14756246 TI - Possible role of dentin matrix in region-specific deposition of cellular and acellular extrinsic fibre cementum. AB - The mechanism whereby a region-specific deposition of the two types of cementum (cellular cementum and acellular extrinsic fibre cementum) is regulated on the growing root surface was tested using bisphosphonate-affected teeth of young rats and guinea pigs. The animals were injected subcutaneously with 8 or 10 mg P x kg body weight(-1) x day(-1) of 1-hydroxyethylidene-1,1-bisphosphonate (HEBP) for 1 or 2 weeks. In rat molars, HEBP prevented mineralization of newly formed root dentin matrix and totally inhibited de novo deposition of acellular extrinsic fibre cementum. Instead, thick cellular cementum was induced on the non mineralized root dentin surface, irrespective of the position of the root. In both animals, cellular cementum was also induced on the non-mineralized surface of root analogue dentin in HEBP-affected incisors, where only acellular extrinsic fibre cementum is deposited under normal conditions. In normal rat molars, dentin sialoprotein (DSP) was concentrated along the dentin-cellular cementum border, but not that of dentin and acellular extrinsic fibre cementum. In HEBP-affected rat incisors, DSP was shown to penetrate through the non-mineralized dentin into the surrounding tissues, but not through the mineralized portions. These data suggest that, at the site of cellular cementum formation, putative inducing factors for cellular cementum might diffuse into the periodontal space through the newly deposited mantle dentin matrix before it is mineralized. At earlier stages of root formation, mantle dentin might mineralize more promptly not to allow such diffusion. The timing of mineralization of mantle dentin matrix might be the key determinant of the types of the cementum deposited on the growing root surface. PMID- 14756247 TI - Possible role of immunocompetent cells and the expression of heat shock protein 25 in the process of pulpal regeneration after tooth injury in rat molars. AB - Recent studies have established that heat shock proteins (HSPs) potentially play a role in immunosurveillance. The purpose of the present study was to clarify the relationship between the chronological changes of immunocompetent cells and the expression of HSP-25 in the process of pulpal regeneration after tooth injury in rat molars by immunocytochemistry for HSP-25 and class II major histocompatibility complex (MHC) antigen. In untreated control teeth, intense HSP 25 immunoreactivity was found in the cell bodies of odontoblasts. Both cavity preparation and tooth replantation caused the degeneration of the odontoblast layer to result in the loss of HSP-25 immunoreactions in the suffered dental pulp at the early stages after tooth injury. Numerous class II MHC-positive cells appeared along the pulp-dentin border and extended their cell processes into the dentinal tubules at 12-24 h after cavity preparation and 3 days after tooth replantation. Newly differentiated odontoblast-like cells with HSP-25 immunoreactivity were arranged at the pulp-dentin border and the class II MHC positive cells retreated towards the subodontoblastic layer by post-operative days 3-5 after tooth injury. Thus, the common cellular events occur during pulpal regeneration following two different experimental injuries. These findings indicate that the time course of changes in the expression of HSP-25 immunoreactivity reflects the degeneration/regeneration process of odontoblasts and that the temporal appearance of the class II MHC-positive cells at the pulp dentin border suggests their participation in odontoblast differentiation as well as in initial defence reactions during the pulpal regeneration process. PMID- 14756248 TI - Expression of extracellular matrix molecules, MMPs and TIMPs in alveolar bone, cementum and periodontal ligaments during rat tooth eruption. AB - Tooth eruption involves extensive degradation and reorganization of extracellular matrix (ECM) components. It is not known how ECM-degrading enzymes are coordinated with each other or how they are regulated in the event. The present study was designed to investigate mRNA expression of inhibitors of metalloproteinases (TIMPs) in comparison with matrix metalloproteinases (MMPs) as well as ECM molecules during rat first molar eruption using in situ hybridization. We also examined how TIMPs are involved in the process of tooth eruption, root formation, cementogenesis and alveolar bone remodelling. Expressions of type-I collagen, osteocalcin, MMPs 2 and 8, and TIMPs 1, 2 and 3 were shown in osteoblasts, osteocytes, cementoblasts, cementocytes and periodontal ligament fibroblasts, and the concomitant high expressions of the ECM molecules, MMPs and TIMPs in alveolar bone, cementum and periodontal ligaments were identified in the middle of first molar eruption. The remodelling of ECM in these periodontal tissues might be regulated through balance among the production of ECM molecules, the degradation of ECM by MMPs and the inhibition of MMPs by TIMPs during tooth eruption. PMID- 14756249 TI - High-resolution electron microscopy of enamel-crystal demineralization and remineralization in carious lesions. AB - Although dental caries destroys dental hard tissues, both demineralization as a destructive process and remineralization as a restorative process take place in carious lesions. These conflicting processes occur constantly either simultaneously or alternately. In crystallographic terms, demineralization is crystal dissolution, and remineralization is restoration of partially dissolved crystals, new crystal formation and growth of surviving crystals. In this review, we employ high-resolution transmission electron microscopy to demonstrate demineralization and remineralization of enamel crystals obtained from cross-cut sections of the c-axes of crystals from carious lesions. PMID- 14756250 TI - Fluoride and apatite formation in vivo and in vitro. AB - In recent years, the biomineralization process has attracted much interest from academics and industries for potential technological application. The rule in biomineralization is to have a variety of interfaces and surfaces which can act as nucleators. The ultimate step in any biomineralization process, i.e. the deposition of mineral, must conform to the driving forces operating on the system. A new paradigm in the assessment of the driving force for biomineralization is that a variety of ions existing in the mineralizing milieu are not a bystander, but are instead an active player that directly regulates the precipitation process and nature of biogenic apatites. Thus, the most putative stoichiometric model of a biomineral is (Ca)(5-x)(Mg)q(Na)u(HPO4)v(CO3)w(PO4)(3 y)(OH,F)(1-z). Fluoride participates in many aspects of calcium phosphate formation in vivo and has enormous effects on its process and on the nature and properties of the final products. In the development of biogenic apatites, fluoride ion in the mineralizing media is supposed to accelerate the hydrolysis of acidic precursor(s) and increase the growth rates by augmenting the driving force for precipitation. Inhibitory activities of ions and molecules are related to their adsorption onto the apatite surfaces. From theoretical and practical points of view, it is of paramount importance to elucidate and predict the effect and outcome of fluoride (accelerator) and inhibitors of biological relevance, because of their use in combination for healthcare in dentistry and medicine, e.g. prevention of dental caries and calculus deposition and in the formulation of antiosteoporosis treatments. PMID- 14756251 TI - Biological characteristics of the junctional epithelium. AB - This review summarizes the biological properties of the junctional epithelium, focusing on its developmental aspects, wide intercellular spaces and desmosomes, dense granules, permeability barrier, phagocytotic activity, adhesive structures and nerve terminals. It also discusses the morphology and functions of long junctional epithelium and peri-implant epithelium. Junctional epithelium is derived from the reduced enamel epithelium during tooth development. Apoptosis occurs in the border between oral and reduced enamel epithelia during tooth eruption. Junctional epithelium expresses a cytokeratin-19 immunoreaction, suggesting that this protein is a consistent differentiation marker. Wide intercellular spaces, which contain neutrophils and nerve endings, are formed as there are fewer desmosomes than in the oral epithelium. Dense, membrane-bound granules in the epithelium might correspond with membrane-coating granules, as revealed by their shape, components and freeze-fracture images. Junctional epithelium with high permeability contains exogenously expressed alpha-defensins, while stratified epithelia contain endogenously expressed beta-defensins. The phagocytotic activity in this epithelium remains unclear. Integrin-alpha6beta4 and laminin-5 form a complex in the tooth surface internal basal lamina. Long junctional epithelium created experimentally attaches to the cementum surface by hemidesmosomes and basal lamina. The peri-implant epithelium differs in proliferation and in adhesive structure from the normal junctional epithelium. In conclusion, wide intercellular spaces and poorly developed desmosomes are closely correlated with a permeable nature. There is still uncertainty over the phagocytotic activity of the epithelium. Integrin-alpha6beta4 and laminin-5 form a significant complex in the internal basal lamina. Junctional epithelium receives a rich sensory nerve and has a high rate of cell turnover. Long junctional epithelium can be produced rapidly during wound healing, due to high proliferative activity. Peri-implant epithelium might be a poorly adhered and permeable epithelium. PMID- 14756252 TI - Electrochemical oxidation of phenolic and other organic compounds at boron doped diamond electrodes for wastewater treatment: effect of mass transfer. AB - The paper presents the results of an experimental study on oxidation at boron doped diamond electrodes (BDD) of some phenolic compounds: phenol (PH), para hydroxibenzoic acid (PHB), cathecole (CT), hydroquinone (HQ) are considered, singularly contained in aqueous solutions or in the presence of glucose (G), which was selected to represent the class of biodegradable compounds. Oxidation of benzoquinone (BQ) and maleic acid (MA), generally detected as intermediates products from phenol degradation, is also investigated. Great attention is paid to verify the feasibility of a selective process in which the oxidation is specifically addressed to the phenolic fraction up to non toxic intermediate products which are more biodegradable than the original phenols. PMID- 14756253 TI - A comparative study on direct and indirect electrochemical oxidation of polyaromatic compounds. AB - This study has been performed to investigate the treatment of an industrial wastewater containing naphthalene- and anthraquinone-sulfonic acids, by direct and indirect electrolyses. The direct electrochemical oxidation has been carried out using boron-doped diamond and lead dioxide anodes, while the indirect electrolyses has been mediated by active chlorine electrogenerated on a platinum anode, or by hydrogen peroxide electrogenerated on a graphite felt cathode. For each type of electrolyses the effects of operating parameters, such as anode material, current density, chloride concentration, ferrous ions concentrations have been also investigated. Measurements of Chemical Oxygen Demand (COD) and colour fading have been used to compare the results of different electrolyses. Experimental data showed that the complete COD and colour removals have been obtained only by direct oxidation or by active chlorine mediated electrolyses. On the contrary using electrogenerated hydrogen peroxide the solution has been presented a residual COD and colour. In particular, it found that faster oxidation rate has been obtained by direct oxidation using a boron-doped diamond anodes at low current density. PMID- 14756254 TI - New sol-gel synthetic route to phospho-olivines as environmentally friendly cathodes for Li-ion cells. AB - A new sol-gel synthetic route was developed to prepare single-phase phospho olivines LiMPO4 (M = Fe or Mn), potential cathode materials for the next generation of Li-ion secondary batteries. Triethyl phosphite was used as phosphate organic precursor, with absolute ethanol as solvent. The sol-gel synthesis ensures homogeneity of the precursors at the nanometric scale and improved reactivity, allowing to obtain very small agglomerates and crystal grain size. Several carbon sources were used in order to improve the electrochemical performances of the samples. Galvanostatic cycling tests of the cathodic materials were made on coin cells with Li metal as anode in order to study the influence of the synthesis and the structural-morphological characteristics on the electrochemical performance of the phosphate/C composite. PMID- 14756255 TI - Use of a standard system to evaluate the matrix effect on the treatment of a solution from atrazine contaminated soils. AB - The influence of humic substances on the electrochemical treatment of solutions resulting from the remediation of atrazine contaminated soil has been investigated. In particular the effect on the hydrogen peroxide production stage and the effect on the hydroxyl radical oxidation treatment have been separately studied. In order to quickly assess the possible inhibition of the removal, an inorganic system has been adopted as a standard system. The results confirm the validity of electrochemical technologies also for the treatment of a real effluent where a matrix effect is expected since the organic matter content affects only the hydrogen peroxide production. PMID- 14756256 TI - Treatment of the solution extracted from metal contaminated soils by reverse osmosis and chemical precipitation. AB - In this study a process for the remediation of soils contaminated by lead or copper is proposed, consisting of the operations in sequence: soil flushing, membrane treatment, acidification, and metal precipitation. Pb(II) and Cu(II) extraction from a synthetically contaminated soil using a 0.05 M EDTA aqueous solution were investigated in column. The metal removal efficiencies and the final soil metal concentration were 98.2% and 37.96 mg/g respectively for lead and 95.4% and 59.20 mg/kg for copper. The extracted solutions were concentrated through a membrane treatment to reduce the water content up to the 75% and to obtain a permeate metal concentration in compliance with the Italian Environmental Regulation. The recovery of the used EDTA from the retentate solution, with recovery yield of at least 85.4%, was also obtained through acidification. Metal precipitation from the filtered solution was then performed according two different methods, achieving metal removal yield of more than 99.4%. PMID- 14756257 TI - Which kind of data (normally distributed, standardized, transformed) for a realistic factor analysis? AB - Taking into account that the problem of the best pretreatment of data for factor analysis has not yet arrived at generally accepted solutions, it has been tried to estimate, by an empiric procedure, the goodness of the results of repeated factor analyses with different pretreatments, conditions and statistical distribution of variables. Sets of multivariate data for river waters have been constructed firstly, after fixing the number and nature of the latent factors corresponding to the sources of pollution. A series of R-mode factor analyses has then been performed on these data, using various pretreatments (autoscaling, logarithmic transformation and their combinations), various factor rotations (rigid and oblique) and methods of computation of significant factors (40 data processing on the whole). Factor analyses have also been performed on real data of the Po river in the Piedmont region. Comparisons between the results obtained by factor analysis and the actual situation of the systems investigated have made it possible to draw some conclusions on how to proceed in order to obtain realistic results with this chemometric technique. PMID- 14756258 TI - Dioxouranium-carboxylate complexes. Formation and stability of acetate species at different ionic strengths in NaCl(aq). AB - The formation and stability of UO2(2+)-acetate complexes was studied potentiometrically, at T = 25 degrees C, at different ionic strength, 0 < I < or = 1 mol dm(-3), in aqueous NaCl solutions. Computer analysis allowed us to find the species UO2(ac)+, UO2(ac)2(0), UO2(ac)3- and UO2(ac)3(OH)2-. The dependence on ionic strength of formation constants was taken into account by using both a simple Debye-Huckel type equation and the SIT (Specific ion Interaction Theory) approach. A critical examination of the present results is given together with a comprehensive analysis of literature data. PMID- 14756259 TI - Equilibria occurring between beryllium (II) and salicylate ions. AB - The complexation equilibria between Be2+ and the hydrogen salicylate (HL-) ions have been studied, at 25 degrees C, by potentiometric measurements with a glass electrode in 3 M NaClO4. The concentrations of metal (CM) and ligand (CL) were varied between 10(-3) and 0.03 M and 2 x 10(-3) and 0.03 M, respectively, while 1 < or = CL/CM < or = 3. The hydrogen ion concentration ranged from 10(-3) to 10( 5.3) M when basic salts start to precipitate. The equilibria can be written in the general form as: pBe2+ + rHL- <==> Be(p)H(-q) (HL)r(2p-r-q) + qH+, log beta(pqr). The experimental data have been explained with the formation of BeHL+ (log beta101 = 1.46 +/- 0.05), BeL (log beta111 = -0.897 +/- 0.018), BeL2(2-) (log beta122 = -3.746 +/- 0.021), Be2(OH)L2- (log beta232 = -5.23 +/- 0.09), Be3(OH)3L3(3-) (log beta363 = -14.39 +/- 0.12). The uncertainties represent 3sigma. The predominant complex in the whole concentration range studied is the uncharged mononuclear species BeL. PMID- 14756260 TI - A new approach to the integrated calibration in flow injection analysis. AB - A new proposal how to perform the analytical procedure according to the integrated calibration method is presented. An original flow injection system has been designed for this purpose. When using only a single standard solution, the measurement information gathered during a single analytical course permits construction of four calibration graphs and calculation of as many as four independent estimations of the analyte concentration in the sample examined. As the calibration method applied integrates the set of standards method and the standard addition method, the analytical estimations may be obtained in both the interpolative and the extrapolative manner and the final result can be effectively verified in terms of accuracy. The system developed was experimentally tested on the example of spectrophotometric determination of chromium. It has been proved to be capable of saving time and reagents as well as providing reliable analytical results. Owing to the instrumental simplicity and analytical efficiency the system is expected to be useful for routine analysis. PMID- 14756261 TI - The acid-base equilibrium constants of some azine compounds in various aqueous organic solvent mixtures. AB - The acid dissociation constants of the protonated form of some azine compounds (acridine, acridine orange and neutral red, BH+) were determined pH-metrically at 25 degrees C and at the constant ionic strength I = 0.1 mol l(-1) (KNO3) in pure water as well as in various aqueous mixtures having different proportions (w/w%) of organic solvents. The organic solvents used are methanol, ethanol (as amphiprotic solvents), N,N-dimethylformamide, dimethylsulfoxide (as dipolar aprotic solvents) and acetonitrile (as a low basic solvent). The results obtained indicated that the pKa values decrease as the content of the organic solvent in the medium is increased. It is deduced that, the major effect responsible for this behaviour is the differences in stabilization of the free base (B) by dispersion forces and of the proton by its interaction with solvent and water molecules in aqueous-organic solvent mixtures (ion-solvent interaction). Moreover, it is concluded that the ability of the solvent to accept hydrogen bond from the protonated form (BH+) contributes significantly to the deprotonation process of the compounds. PMID- 14756262 TI - Mercury in the epiphytic lichen Parmelia caperata from Liguria (north-west Italy). PMID- 14756263 TI - Prediction of 3-yr cadaveric graft survival based on pre-transplant variables in a large national dataset. AB - Pre- and post-transplant predictive factors of graft survival for optimal and expanded criteria grafts have been studied in the past. The goal of our study was to evaluate the recent large set of United Network of Organ Sharing records (1990 1998) to generate a prediction algorithm of 3-yr graft survival based on pre transplant variables alone. The dataset of patients with end-stage renal disease and cadaveric kidney or kidney-pancreas transplantation (1990-1998) used in the study consisted of 37,407 records. Logistic regression (LM) and a tree-based model (TBM) were used to identify predictors of 3-yr allograft survival and to generate prediction algorithm. Donor and recipient demographic characteristics (age, race, and gender) and body mass index showed non-linear, while human leukocyte antigen match showed strong linear relationships with 3-yr graft survival. Prediction of the probability of graft survival from the model, achieved a good match with the observed survival of the separate dataset, with a correlation of r = 0.998 for LM and r = 0.984 for TBM. The positive predictive value (PV) of allograft survival with LM and TBM was 76.0% and the negative PV was 63 and 53.8% for LM and TBM, respectively. Both LM and the TBM can potentially be used in clinical practice for long-term prediction of kidney allograft survival based on pre-transplant variables. PMID- 14756264 TI - Mini incision live donor nephrectomy: an optimal approach for the developing countries. AB - OBJECTIVE: Laparoscopic donor nephrectomy (LD) is rapidly gaining popularity, however, this may not be affordable by donors in many developing countries because of its high cost. We describe our mini flank incision (MD) donor nephrectomy technique and its outcome. METHODS: A 7-10-cm subcostal rib sparing transverse incision was given 2 cm lateral to the tip of the 12th rib, towards the lateral border of rectus muscle. All dissections were performed with help of long retractors and instruments, vessels were transfixed and cut. In last 45 cases, vessels were clipped with Liga or Weck clips. Donors and recipients outcome was analysed. RESULT: From January 2000 to December 2002 a total of 148 patients underwent donor nephrectomy by mini incision technique. Mean patient age was 44.8 +/- 7.3 yr (range 20-70 yr). Nephrectomies were performed in 115 patients on the left side and in 33 cases on the right side. The mean incision length was 9.1 +/- 1.8 cm (range 7-10 cm). Mean operative time was 105 +/- 10.5 min (70-130 min). Mean analgesic (Tramadol) requirement was 205 +/- 52 mg; postoperative hospital stay was 2.2 +/- 0.5 d. Twelve per cent patients developed fever and 4% had superficial wound infection in postoperative period. Three patients required blood transfusion. Mean convalescence period was 22 +/- 2.8 d. CONCLUSION: Extrapleural, extraperitoneal, subcostal mini incisions live donor nephrectomy is a relatively safe procedure with low morbidity. This technique has a shorter hospital stay, early convalescence and better cosmesis. It is cost effective and is an ideal substitute for the developing country. PMID- 14756265 TI - Remaining experiences of living kidney donors more than 3 yr after early recipient graft loss. AB - Living kidney donor programs, based on willingness among family members and close relatives to donate, have made it possible to perform a satisfactory number of kidney transplantations. Early graft loss in the recipient may occur and it is not known if such an event will result mainly in acute, rather transient, emotional reactions or if long-lasting reactions may be evoked in the living kidney donor. The aim of the present study was to assess and describe the remaining experiences of donors (n = 10) more than 3 yr after early recipient graft loss or death of the recipient. A phenomenographic, interview-based research approach was used. Five different fields or domains were identified: (i) the decision to donate; (ii) the information provided; (iii) care received at the time of donation; (iv) responses at graft failure; and (v) concerns remaining at the time of the interview. All donors expressed that they had volunteered to donate and that no stress had been put on them. The information given prior to and in connection with the donation procedure was deemed insufficient but all donors were satisfied with the medical care provided in connection with the nephrectomy and in the immediate post-operative period. Graft failure was immediately accepted on the intellectual level by nine of 10 donors but still evoked emotional reactions and responses included a wish that continuing contact with the transplant staff had been provided. The present interview-based study shows that it is of importance that the donor is thoroughly informed about all donor as well as recipient-related factors including the potential risk of recipient graft failure. In case of graft failure, or the death of the recipient, the transplant unit staff members should offer contact for discussions of medical matters as well as for psychosocial support. In individual cases it may be necessary to maintain such a supportive contact channel for a prolonged period of time. PMID- 14756267 TI - Long-term outcome of kidney transplant using non-heart-beating donor: multicenter analysis of factors affecting graft survival. AB - This multicenter study was retrospectively evaluated for the predictive factors affecting the long-term graft survival of a kidney transplant from a non-heart beating donor (NHBD). PATIENTS AND METHOD: A total of 706 patients received transplants from NHBD in 11 centers between 1986 and 2000 and the results were entered into the analysis. The patients were treated with cyclosporine- or tacrolimus-based immunosuppressive therapy. Graft survival was calculated by the Kaplan-Meier method. Factors selected for univariate analysis were donor age, and acute early and acute late rejection. Hypertension (HT), hyperlipidemia (HL), and diabetes mellitus were also analyzed in 638 recipients whose graft survived for more than 1 yr. RESULTS: In the cases using NHBD, graft survival for 1, 5, and 10 yr was 87, 69, and 53%, respectively. Donor age of over 55 yr, acute early and late rejection, post-transplant HT and diabetes at the first post-operative year were shown to be significantly harmful on long-term graft survival. For longer graft survival in NHBD kidney transplantations, reducing acute rejection, and controlling blood pressure and sugar are crucial. PMID- 14756266 TI - Pharmacokinetics of daclizumab and mycophenolate mofetil with cyclosporine and steroids in renal transplantation. AB - Daclizumab and mycophenolate mofetil (MMF) decrease the incidence of acute allograft rejection. This double-blind, randomized, placebo-controlled trial was performed primarily to assess the pharmacokinetics of MMF in an immunosuppressive regimen incorporating daclizumab. At five centers, 75 renal transplant recipients were randomized 2:1 to receive either daclizumab 1 mg/kg or placebo pre transplantation and every other week, for a total of five doses. All patients received cyclosporine, steroids, and MMF. Levels of mycophenolic acid (MPA), its glucuronide metabolite, and daclizumab were measured after dosing on days 28 and 56. Safety parameters evaluated included: adverse events, laboratory abnormalities, infections, patient/graft survival, incidence of lymphoproliferative disorders, and incidence of acute rejection at 12 months. The concomitant administration of daclizumab and MMF had no effect on the pharmacokinetics of MPA: AUC(0-8) values (microg h/mL +/- SD) on day 28 were 30.1 +/- 13.3 for daclizumab-treat patients vs. 31.1 +/- 12.4 for placebo and on day 56, 37.7 +/- 18.2 for daclizumab-treated patients vs. 35.7 +/- 14.0 for placebo. Adverse events were similar between the two groups. Acute rejection at 12 months occurred in 14% of patients receiving daclizumab and 20% of patients receiving placebo. The coadministration of daclizumab did not result in a pharmacokinetic interaction with MPA, the active metabolite of MMF. PMID- 14756268 TI - Skin infections in renal transplant recipients and the relation with solar ultraviolet radiation. AB - BACKGROUND: Ultraviolet radiation (UVR) is an important risk factor for skin cancer in transplant recipients. In view of the potential suppressive effect of UVR on host resistance it was examined whether exposure to UVR was also associated with the occurrence of various skin infections. METHODS: In a cohort of renal transplant recipients (n = 137), lifetime exposure was assessed by means of a retrospective questionnaire on cumulative sunlight exposure. Diagnosed skin infections since renal transplantation were extracted from the patient's medical charts. Season of diagnosis was regarded as indicative of short-term exposure. RESULTS: In comparison with winter a high rate of herpes simplex infections was found in spring [rate ratio (RR) = 4.09, 95% confidence interval (CI) 1.2-14.5], and high rates of herpes zoster infections (RR = 1.6, 95% CI: 0.8-3.5) and fungal/yeast infections in summer (RR = 2.1, 95% CI: 1.3-3.4). A higher lifetime exposure (RR = 2.31, 95% CI: 1.04-5.1) and a greater cumulative number of reported sunburns (RR = 2.3, 95% CI: 1.1-5.1) were independently associated with a higher risk of bacterial infections. CONCLUSIONS: The seasonal association with the occurrence of clinical herpes infections indicates an effect of short-term UVR. Our data suggest that the number of sunburn episodes in the past is also relevant for the susceptibility to certain skin infections. PMID- 14756269 TI - Endothelial function is more impaired in hemodialysis patients than renal transplant recipients. AB - BACKGROUND: Endothelial dysfunction (ED) is a common precursor and denominator of cardiovascular events including development of atherosclerosis. In this cross sectional, controlled study, we aimed to investigate ED measured by ischemia induced forearm vasodilatation in chronic hemodialysis (HD) patients and renal transplant recipients (rTX). PATIENTS AND METHODS: Thirty-nine HD patients, 39 rTX and 38 normotensive healthy controls were included. There was no difference in age and gender distribution among the study groups. The mean time spent on dialysis and transplantation were 74 +/- 46 and 68 +/- 39 months. Serum high sensitive C-reactive protein (hs-CRP) and plasma fibrinogen levels were measured. Endothelium dependent post-ischemic vasodilatation of brachial artery was used to evaluate ED. RESULTS: The hs-CRP and plasma fibrinogen levels were significantly increased in HD patients when compared with rTX. On high resolution ultrasonographic examination, post-ischemic vasodilatation values in HD patients (9.55 +/- 6.47%) were significantly lower than rTX (14.39 +/- 8.06%, p = 0.007) and controls (20.42 +/- 6.10%, p < 0.001). Renal transplant recipients also had significantly lower post-ischemic vasodilatation values than controls (p = 0.001). The hs-CRP levels were negatively correlated with endothelium-dependent dilatations in TX (r = -0.59, p = 0.001), however, this correlation was not detected in HDp. CONCLUSION: Patients with end-stage renal disease have ED. Endothelial function is more impaired in HD patients than rTX. Different mechanisms might be responsible for ED in HD patients and rTX. PMID- 14756270 TI - Hypoglycaemia after pancreas transplantation: usefulness of a continuous glucose monitoring system. AB - BACKGROUND: After pancreas transplantation (PTx) some patients report occasional symptoms of hypoglycaemia and at times, serious hypoglycaemia. Continuous blood glucose monitoring (CBGM) allows determination of the daily glucose profile and detection of unrecognized hypoglycaemia. The aims of our study were to determine the incidence of hypoglycaemia in PTx and evaluate whether the use of CBGM helps to detect unrecognized nocturnal hypoglycaemia. PATIENTS AND METHODS: We studied 12 patients (six males) with normal functioning PTx and kidney transplantation for more than 3 yr, with systemic drainage of endocrine secretion and stable immunosuppression. A 24-h CBGM using a microdialysis technique (GlucoDay, A. Menarini Diagnostics, Florence, Italy) was performed in all the patients. RESULTS: Three patients had asymptomatic recorded glucose levels below 3.3 nmol/L during the nocturnal period (01:00-07:00 hours) with the glucose levels during these episodes being 2.6, 2.5 and 2.5 nmol/L, and the duration of nocturnal hypoglycaemia being 27, 62 and 93 min, respectively, rising spontaneously without intervention. Patients with hypoglycaemia presented lower glycosylated haemoglobin levels when compared with those not presenting hypoglycaemic episodes, although basal glucose and insulin levels and insulin antibody titres were similar. In one of the three patients presenting hypoglycaemia CBGM was re evaluated after including an extra snack at bedtime, with subsequent normalization of the blood glucose profile being observed. CONCLUSION: Unrecognized nocturnal hypoglycaemia is relatively frequent in patients with PTx and 24-h CBMG may be useful to detect these episodes. PMID- 14756271 TI - Induction therapy by anti-thymocyte globulin (rabbit) in renal transplantation: a 1-yr follow-up of safety and efficacy. AB - BACKGROUND: Two hundred and forty cadaveric renal transplant recipients given anti-thymocyte globulin (Thymoglobulin) as induction immunotherapy were followed up prospectively to review safety and efficacy. METHODS: The median number of infusions was 10 [2-21] with a cumulative dose of 8.8 mg/kg [2.0-23.2 mg/kg]. During the fortnight following transplantation, 231 patients (96%) received a calcineurin inhibitor; all patients were given steroids and azathioprine or mycophenolate mofetil. At 1 yr, 60% of patients were on tripletherapy, 38% on bitherapy, and 2% on monotherapy; 20% had discontinued steroids. RESULTS: Tolerance was excellent with no cases of anaphylaxis. The commonest adverse event was fever (55%). Eighteen patients developed serum sickness on median day 11 [10 14]. Seven patients had thrombocytopenia; six patients had severe neutropenia. All of these adverse events recovered spontaneously. The overall incidence of delayed graft function was 24%. At 1 yr patient and graft survival were 98 and 95%, respectively, and creatinine was 135 +/- 43 micromol/L. Clinically suspected and biopsy-proven acute rejection were observed in 65 patients (27%) and 34 patients (14%), respectively. There were 62 non-cytomegalovirus (CMV) infections (two fatal) and 81 episodes of CMV infections. Eight malignancies were reported; two possibly related to immunosuppression. CONCLUSIONS: These results demonstrate that anti-thymocyte globulin has a safety profile with good tolerability and excellent efficacy. PMID- 14756272 TI - Cyclosporin A monitoring by 2-h levels: preliminary target levels in stable pediatric kidney transplant recipients. AB - Clinical trials in adults have shown that management of transplanted patients with cyclosporin A (CsA) 2-h levels (C2) lead to superior outcome compared with monitoring of 12-h trough levels (C0). In both adults and children, C2 levels enabled a better estimation of the area under the curve concentration than C0 levels. Therefore, it can be suspected that C2 monitoring might also lead to a better outcome in children. Until now C2 target levels for children have not been defined. We measured C2 levels in 101 stable pediatric kidney recipients with a minimum time of 1 yr after transplantation. C2 levels were compared with changes in glomerular filtration rate (GFR) 6 months later. Median C2 levels in children after renal transplantation were 714 ng/mL (95% confidence interval 654-774). Patients with C2 levels below 750 ng/mL had a significantly higher percentage of decline in GFR than patients with C2 levels above 750 ng/mL (p < 0.05). In children with C2 levels below 500 ng/mL three acute rejections occurred in comparison with no rejection in the remaining patients (p < 0.05). We conclude that the lower C2 target level should be above 750 ng/mL in stable pediatric transplant recipients. An upper target level above 1000 ng/mL should be avoided. The question, whether C2 monitoring in pediatric kidney recipients is superior to C0 monitoring, is yet to be answered. PMID- 14756273 TI - Chemoembolization followed by orthotopic liver transplant for epithelioid hemangioendothelioma. AB - Hepatic epithelioid hemangioendothelioma (HEHE) is a rare liver tumor with an indolent course relative to other hepatic malignancies. Over the past two decades, primary treatment for these lesions has been defined as resection for localized disease, or transplantation for diffuse and multifocal tumors. No published report to date has described effective pre- or post-operative adjuvant treatment for this disease. In this report, we present the first case of HEHE effectively managed with chemoembolization followed by transplantation, documenting objective tumor response to embolization. Furthermore, diagnosis for this lesion can easily be mistaken, directing management in erroneous directions. This case illustrates diagnostic pitfalls affiliated with the work-up of this tumor. PMID- 14756275 TI - Percutaneous penetration of inorganic mercury from soil: an in vitro study. PMID- 14756274 TI - Efficacy of combined lamivudine and adefovir dipivoxil treatment for severe HBV graft reinfection after living donor liver transplantation. AB - Resistance to lamivudine and hyperimmune globulin (HBIG) may cause severe graft reinfection with progression to fulminant hepatic failure in liver transplant recipients. In this report, we describe the clinical course of a patient with perinatally acquired chronic hepatitis B virus (HBV) infection and hepatocellular carcinoma who developed severe fibrosing cholestatic hepatitis after living donor liver transplantation because of the emergence of lamivudine and HBIG-resistant chronic hepatitis B. Immunohistochemistry demonstrated that more than 30% of hepatocytes stained positively for hepatitis B core antigen. Hepatitis B virus sequence analysis revealed several mutations in the polymerase gene (L528M, M552I, M552V) as well as in the surface gene region encoding the immunogenic major hydrophilic loop of the small surface protein (G130N, M133T, D144G). The amino acid exchange at codon 144 has already been described to escape neutralization by HBIG. Combined treatment with lamivudine and adefovir dipivoxil (ADV) was associated with a dramatic biochemical, virological and clinical response with resolution of jaundice, ascites, peripheral edema and pleural effusions. Serum bilirubin normalized, HBV DNA levels significantly decreased and liver biopsy was remarkable for the absence of viral protein. These results indicate that ADV may provide a sustained rescue treatment for aggressive courses of HBV graft reinfection in liver transplant recipients. PMID- 14756276 TI - Toxicity issues associated with geogenic arsenic in the groundwater-soil-plant human continuum. PMID- 14756277 TI - Cadmium exposure of the Greek population. PMID- 14756278 TI - QSARs for predicting the toxicity of mixtures containing polar narcotic chemicals. PMID- 14756279 TI - QSAR for prediction of joint toxicity of substituted phenols to tadpoles (Rana japonica). PMID- 14756280 TI - Vineyard pesticides and their effects on invertebrate biomarkers and bioindicator species in New Zealand. PMID- 14756281 TI - Tolerance: a useful biological parameter for identifying contaminated sites. PMID- 14756282 TI - Plasmid-encoded heavy metal resistance in Pseudomonas sp. PMID- 14756283 TI - Histopathological changes in the testis of the Sprague Dawley rat following orally administered manganese. PMID- 14756284 TI - Single and binary-combined toxicity of methamidophos, acetochlor and copper acting on earthworms Esisenia foelide. PMID- 14756285 TI - Toxicity of copper on four Chilean marine mussels. PMID- 14756286 TI - Nickel induced toxic effects and bioaccumulation in the submerged plant, Hydrilla verticillata (L.F.) Royle under repeated metal exposure. PMID- 14756287 TI - Removal of copper toxicity by zeolite in Java tilapia Oreochromis mossambicus (Peters). PMID- 14756288 TI - Hematological responses in a freshwater fish Channa punctatus due to fenvalerate. PMID- 14756289 TI - Sublethal effects of tannery effluent on some hematological indices and growth of Clarias gariepinus (Teugels). PMID- 14756290 TI - Toxicity and residue studies of fenvalerate to the freshwater fish Channa punctatus (Bloch). PMID- 14756291 TI - Comparison of matrix modifiers in the determination of cadmium and lead in industrial waste water plants around Cairo by graphite furnace atomic absorption spectrophotometry. PMID- 14756292 TI - Hydro-geochemical behavior of bicarbonate and sulfate ions leaching from a sulfide-poor silver mine in central Mexico: potential indicator of acid mine drainage. PMID- 14756293 TI - Pesticides and heavy metal distribution in southern Dead Sea basin. PMID- 14756294 TI - Lead levels in ambient air and blood of pregnant mothers from the general population of Lucknow (U.P.), India. PMID- 14756295 TI - Persistent organochlorine pesticides in coastal sediments from Petacalco Bay, Guerrero, Mexico. PMID- 14756296 TI - Polycyclic aromatic hydrocarbons and aliphatics in the coral reef skeleton of the Egyptian Red Sea coast. PMID- 14756297 TI - Market basket survey for lead, cadmium, copper, chromium, nickel, and zinc in fruits and vegetables. PMID- 14756298 TI - Accumulation of copper and cadmium in small and large Nile tilapia Oreochromis niloticus. PMID- 14756299 TI - Genotypic differences in effect of Cd on photosynthesis and chlorophyll fluorescence of barley (Hordeum vulgare L). PMID- 14756300 TI - Nitrous oxide emission and urease activity in wheat. PMID- 14756301 TI - Effect of zinc-cadmium interactions on the uptake of zinc and cadmium by winter wheat (Triticum aestivum) grown in pot culture. PMID- 14756302 TI - Isolation and identification of a super strong plant promoter from cotton leaf curl Multan virus. AB - The activity of the C1 and the V1 gene promoter of cotton leaf curl Multan virus (CLCuMV) was investigated in transgenic plants with the gus gene as a reporter gene. Quantitative GUS activity analysis of the transgenic plant leaves showed the average activity of the CLCuMV C1 gene promoter was 3- to 5-fold higher than that of the CaMV 35S promoter, with maximal expression being 10-fold higher. CLCuMV V1 gene promoter activity was only about 1/10th that of the CaMV 35S promoter in the absence of trans-activator C2. Histochemical GUS staining of the transgenic plants indicated that the CLCuMV C1 gene promoter was active in leaves, stems, roots and almost all reproductive organs. Functional analysis of promoter 5'-deletion series indicated that promoter activity of a 257 nucleotide fragment (-257 to the transcription initiation site) and a 241 nucleotide fragment (-241 to the transcription initiation site) were 5-fold and 2-fold stronger than that of the full-length CLCuMV C1 promoter respectively. These results demonstrate that the CLCuMV C1 promoter is a super-strong near constitutive promoter in plants and has great application potential for plant genetic engineering studies. PMID- 14756303 TI - Characterization of all the subunits of replication factor C from a higher plant, rice (Oryza sativa L.), and their relation to development. AB - Replication factor C (RFC), which is composed of five subunits, is an important factor involved in DNA replication and repair mechanisms. Following previous studies on the RFC3 homologue from rice (Oryza sativa L. cv. Nipponbare) (OsRFC3), we succeeded in isolating and characterizing one large and three small subunits of RFC homologues from the same rice species and termed them OsRFC1, OsRFC2, OsRFC4 and OsRFC5. The plant was found to have all RFC subunits known in yeasts, human and other eukaryotes. The open reading frames of OsRFCs encoded a predicted product of 1021 amino acid residues with a molecular mass of 110.8 kDa for OsRFC1, 339 amino acid residues with a molecular mass of 37.4 kDa for OsRFC2, 335 amino acid residues with a molecular mass of 36.8 kDa for OsRFC4, and 354 amino acid residues with a molecular mass of 40.5 kDa for OsRFC5. All the OsRFC subunits have highly conserved amino acid motifs among RFC proteins, RFC box, and an unrooted phylogenetic tree shows each OsRFC subunit belongs to each RFC subunit group. These subunits showed differences in their expression patterns among tissues. The transcripts of OsRFCs were expressed strongly in the proliferating tissue, the shoot apical meristem (SAM), and very weakly in the mature leaves which have no proliferating tissues. However, in young leaves and flag leaves, tissue-specific expression of OsRFC3 and OsRFC4 was shown. On the other hand, cell cycle arrest by cell cycle inhibitors resulted in significant differences in OsRFC expression patterns. These results suggest the functional differences of each OsRFC subunit in tissues and the plant cell cycle. The roles of these molecules in plant DNA replication and DNA repair are discussed. PMID- 14756304 TI - Characterization of two phosphate transporters from barley; evidence for diverse function and kinetic properties among members of the Pht1 family. AB - Putative phosphate transporters have been identified in a barley (Hordeum vulgare L.) genomic library by their homology to known phosphate transporters from dicot species. The genes designated HORvu;Pht1;1 and HORvu;Pht1;6 encode proteins of 521 and 535 amino acids respectively with 12 predicted membrane-spanning domains and other motifs common to the Phtl family of phosphate transporters. HORvu;Pht1;1 is expressed exclusively in roots and is strongly induced by phosphate deprivation. HORvu;Pht1;6 is expressed in the aerial parts of the plant with strongest expression in old leaves and flag leaves. In situ hybridization showed that HORvu;Pht1;6 is expressed in the phloem of vascular bundles in leaves and ears. In order to study the biochemical properties of HORvu;Pht1;1 and HORvu;Pht1;6, the genes were expressed in transgenic rice (Oryza sativa L.) plants under the control of the rice actin promoter and suspension cell cultures were generated. Cells derived from transgenic plants were able to take up phosphate at a much higher rate than control cells, demonstrating that both genes encode functional phosphate transporters. The estimated Km for phosphate for cells expressing HORvu;Pht1;1 was 9.06 +/- 0.82 microM, which is characteristic of a high-affinity transporter. The rate of phosphate uptake decreased with increasing pH, suggesting that HORvu;Pht1;1 operates as a H+/H2PO4(-) symporter. In contrast, the estimated Km for phosphate for cells expressing HORvu;Pht1;6 was 385 +/- 61 microM, which is characteristic of a low-affinity transporter. Taken together, the results suggest that HORvu;Pht1;1 functions in uptake of phosphate at the root surface, while HORvu;Pht1;6 probably functions in remobilization of stored phosphate from leaves. PMID- 14756305 TI - A RING-H2 zinc-finger protein gene RIE1 is essential for seed development in Arabidopsis. AB - RING zinc-finger proteins play important roles in the regulation of development in a variety of organisms. In the plant kingdom, few genes encoding RING zinc finger proteins have been documented with visible effects on plant growth and development. A novel gene, RIE1, encoding a RING-H2 zinc-finger protein was identified in Arabidopsis thaliana and is characterized in this paper. RIE1 encodes a predicted protein product of 359 amino acids residues with a molecular mass of 40 kDa, with a RING-H2 zinc-finger motif located at the extreme end of the C-terminus. Characterization of a Dissociation (Ds) insertion line (SGT4559) and a T-DNA insertion line (SRIE1) demonstrated that disruption of RIE1 is embryo lethal. SGT4559 heterozygous plants produced seeds with embryo development arrested from globular to torpedo stages. Some mutant seeds were rescued by embryo culture, and the mutant (rie1) plants seemed to grow normally compared to wild-type plants, except that the mutants produced only abnormal seeds. However, RIE1 was expressed in different tissues throughout the whole plant as revealed by northern blot analysis and gene fusion assay of RIE1 promoter with the beta glucuronidase (GUS) gene. Our results indicated that RIE1 plays an essential role in seed development. PMID- 14756306 TI - Fructose-2,6-bisphosphate contents were increased in response to salt, water and osmotic stress in leaves of Bruguiera gymnorrhiza by differential changes in the activity of the bifunctional enzyme 6-phosphofructo-2-kinase/fructose-2,6 bisphosphate 2-phosphatase. AB - The steady-state level of 6-phosphofructo-2-kinase/fructose-2,6-bisphosphate 2 phosphatase (F6P2K/F26BPase) transcript has been found to be raised in the mangrove Bruguiera gymnorrhiza treated with 500 mM NaCl for 6 h. In the present study, we assayed both F6P2K/F26BPase activity and fructose-2,6-bisphosphate (F26BP) contents in leaves of salt- and water-stressed B. gymnorrhiza. In the plants treated with 500 mM NaCl, no increase in transcript level was observed after 1 day of treatment, while both the ratio between F6P2K and F26BPase activity (K/P ratio) and leaf F26BP level were about two-fold higher than in control plants. Several water stress-associated treatments, including 500 mM NaCl treatment for 6 h, 1 M mannitol treatment for 6 h and dehydration treatment, resulted in increases in leaf F26BP level as compared with water-grown plants. The raised levels of F26BP in osmotically stressed plants treated with NaCl and mannitol were accompanied with increased transcript levels and subsequent increases in both F6P2K and F26BPase activities, while the increase in F26BP levels in dehydrated plants was attributed to an increase in K/P ratio without an increase in transcript levels. These results suggest that, although both treatments resulted in increases in F26BP levels, B. gymnorrhiza differentially responds to osmotic stress and water stress. PMID- 14756308 TI - Post-transcriptional regulation of expression of the Bronze2 gene of Zea mays L. AB - The glutathione S-transferase encoded by Bronze2 performs the last genetically defined step in maize anthocyanin biosynthesis, being required for pigment sequestration into vacuoles. The Bz2 primary transcript contains a single intron; in maize leaves both spliced and unspliced Bz2 transcripts are usually present and are predicted to encode 26 and 14 kDa proteins, respectively. To increase understanding of the role and regulation of Bz2 transcript splicing, we examined Bz2 expression during development in transgenic maize plants expressing a 35S:Bz2 (35S:mycBz2i) gene and, by transient expression analysis, in Black Mexican Sweet maize protoplasts. We show here that the gene is expressed in diverse tissues that lack functional copies of one or both transcription factors regulating anthocyanin synthesis, that transcript levels are much higher when the R/B plus C1/Pl transcription factors are present, and that the splicing decision depends on local sequence context. The predicted 14 kDa protein was never detected indicating that unspliced transcripts are likely to be non-coding. The native 26 kDa BZ2 protein is loosely membrane-bound, but it was detectable only in tissues accumulating anthocyanin. Consequently, BZ2 accumulation appears to be limited by stringent post-transcriptional regulation. PMID- 14756307 TI - Sensitization of defense responses and activation of programmed cell death by a pathogen-induced receptor-like protein kinase in Arabidopsis. AB - During the search for potential target genes of WRKY DNA-binding transcription factors, we have previously identified four pathogen-induced Arabidopsis genes (CRK5, CRK6, CRK10 and CRK11) encoding receptor-like protein kinases (RLKs) containing novel cysteine-rich repeats in their extracellular domains. In the present study, we transformed Arabidopsis plants with the RLK genes under control of the constitutive CaMV 35S promoter or a steroid-inducible Ga14 promoter. Expression of CRK5, but not the three other RLK genes, resulted in significant alterations in defense responses and leaf growth in transgenic plants. In transgenic plants harboring the 35S::CRK5 construct, significantly elevated and constitutive expression of CRK5 correlated with enhanced leaf growth and increased resistance to the bacterial pathogen Pseudomonas syringae. The enhanced disease resistance in the transgenic plants was associated with more rapidly induced expression of the PR1 gene after pathogen infection. In transgenic plants transformed with CRK5 under control of the steroid-inducible promoter, expression of the transgene was induced at relatively high levels after the steroid application and this induced expression of CRK5 triggered hypersensitive response like cell death. Induced CRK5 expression also activated cell death in the npr1, ndr1 and eds1 mutants and in the transgenic nahG plants that fail to accumulate salicylic acid. Thus, the novel RLK is capable of activating multiple distinct defense responses depending on the manner and/or the levels of its over expression in transgenic plants. PMID- 14756309 TI - Antisense-mediated down-regulation of putrescine N-methyltransferase activity in transgenic Nicotiana tabacum L. can lead to elevated levels of anatabine at the expense of nicotine. AB - Nicotiana tabacum L. produces a number of pyridine alkaloids, with nicotine representing the major component and anatabine comprising most of the remainder of the alkaloid fraction. An antisense approach was used here to down-regulate activity of the important enzyme putrescine N-methyltransferase (PMT) in transformed roots of this species to determine effects upon alkaloid metabolism. Transformed root lines were produced that contained markedly reduced PMT activity, with a concomitant reduction in nicotine content compared to controls. No negative effects upon growth were observed. Several antisense-PMT transformed root lines, and also leaf tissues of regenerated transformed plants, showed a substantial increase in anatabine content relative to controls. Northern hybridization experiments indicated that the antisense-PMT manipulation had little or no effect upon the transcript levels of other genes encoding enzymes involved in alkaloid metabolism, including quinolinate acid phosphoribosyltransferase (QPT). The latter enzyme plays a key role in regulating the synthesis of nicotinic acid which supplies the pyridine ring necessary for both nicotine and anatabine synthesis. We suggest that elevated anatabine levels in antisense-PMT lines are a direct consequence of a relative oversupply of nicotinic acid which, in the absence of adequate levels of 1-methyl-delta(1) pyrrolinium cation (the ultimate product of PMT activity), is used to synthesise anatabine directly. As is discussed, no naturally occurring species or varieties of Nicotiana are known that typically produce high levels of anatabine in root or leaf tissues, meaning that the antisense PMT transgenics produced in this study have no natural counterpart. These experiments thus represent an example of metabolic engineering of plant pyridine metabolism, via antisense down-regulation of gene expression in a contributing pathway leading to secondary metabolite biosynthesis. PMID- 14756310 TI - A novel male-sterile mutant of Arabidopsis thaliana, faceless pollen-1, produces pollen with a smooth surface and an acetolysis-sensitive exine. AB - A mutant exhibiting conditional male sterility, in which fertility was restored under conditions of high humidity, was identified in T-DNA tagged lines of Arabidopsis thaliana. Scanning electron microscopy (SEM) demonstrated that the pollen surface was almost smooth and the reticulate pattern not prominent. Thus, the mutant was named faceless pollen-1 (flp1). Transmission electron microscopy (TEM) revealed that the smooth appearance was due to tryphine filling in the exine cavities and covering the pollen surface. The lipid droplets in the tryphine of mutant pollen were smaller and more numerous than those of the wild type. SEM analysis also demonstrated that pollen exine was easily damaged by acetolysis, suggesting that a component of exine, sporopollenin, was defective in the mutant. In addition, the stems and siliques had reduced amounts of wax crystals. A predicted amino acid sequence of the cDNA that corresponded to the tagged gene, fip1, showed sequence similarity to proteins involved in wax biosynthesis. The FLP1 protein is likely to play a role in the synthesis of the components of tryphine, sporopollenin of exine and the wax of stems and siliques. PMID- 14756311 TI - Profiling ethylene-regulated gene expression in Arabidopsis thaliana by microarray analysis. AB - Ethylene-regulated gene expression in leaves of Arabidopsis thaliana was investigated with an expressed sequence tag-based microarray containing about 6000 unique genes. Comparing expression profiles of the ethylene-insensitive mutant etr1-1, the ethylene-constitutive mutant ctr1-1, ethylene-treated wild type and untreated wild-type plants identified ca. 7% of the investigated genes as ethylene-regulated. Exogenous ethylene treatment and ctr1-1 had similar changes in gene expression, but differences were noted. Ethylene-regulated genes involved in its own biosynthesis and signal transduction pathway were identified. A large number of transcription factors and some putative signaling components were highly regulated by ethylene. Chloroplast structural protein and photosynthetic genes were generally down-regulated. Ethylene appeared to regulate other primary metabolic genes. Plant defense and PR protein genes were differentially regulated, with some genes within this class highly up-regulated. Other ethylene-regulated genes identified were known sugar-, auxin-, wounding- and jasmonic acid-related genes, suggesting the existence of coordinated interactions between ethylene and other hormonal and defense signaling pathways. Although hundreds of potentially important transcriptome changes were identified, the functions of many ethylene-regulated genes remain unknown. PMID- 14756312 TI - Resources for targeted insertional and deletional mutagenesis in Arabidopsis. AB - The maize transposons Activator (Ac) and Dissociation (Ds) are active in many monocots and dicots, including Arabidopsis. We describe a new Ac-derived transposon construct, designated the Ds-loxP T-DNA, which can be used for both insertional and deletional mutagenesis. There are loxP sites in both orientations on both the transposon and the donor site T-DNA and an arrangement of marker genes that permits selection of transposition events, as well as deletions and inversions extending from the donor site to a transposon reinserted on either side of it. We show that Cre-mediated deletions and inversions occur at a high frequency. The tendency of Ac-Ds transposons to reinsert near the donor site can be used to target both insertional and deletional mutagenesis, but efficient exploitation of this property requires a library of mapped marked donor sites distributed in the genome. We have created a population of independent Ds T-DNA transformants and we have mapped an initial set of 75 Ds T-DNA integration sites. We assessed the potential efficiency of targeted mutagenesis by detecting Ds reinsertion events at several loci over a 400 kb interval from each of two donor sites with different Ds T-DNA constructs. The distribution of reinsertion sites is similar around the two tested loci, with roughly 10, 4, and ca. 1% of reinsertions detected within 1-2 kb of sites 10, 100, and 200-400 kb from the donor site, respectively. To facilitate the use of this targeted mutagenesis system. we have constructed a searchable database of the mapped Ds T-DNA integration sites. PMID- 14756313 TI - A 2.5-kb insert eliminates acid soluble invertase isozyme II transcript in carrot (Daucus carota L.) roots, causing high sucrose accumulation. AB - The predominant storage carbohydrates of mature carrot (Daucus carota L.) storage roots typically are the free sugars glucose and fructose. This trait is conditioned by the Rs allele. A naturally occurring recessive mutation, rs/rs, conditions a shift from these reducing sugars to sucrose. RT-PCR and sequencing revealed a unique 2.5 kb insert in the first and largest intron near the 5' end of the acid soluble invertase isozyme II gene of rs/rs carrots. This insert was not totally spliced out during mRNA processing. While the wild-type acid-soluble invertase isozyme II transcript (ca. 2 kb) was detected in Rs/Rs roots and leaves, none was observed in rs/rs roots throughout development. RT-PCR of rs/rs leaves revealed two novel transcripts (2.7 kb and 3.2 kb). A comparison of enzyme activity between the near-isogenic Rs/Rs and rs/rs carrot lines revealed very low acid-soluble invertase activity in rs/rs roots whereas neutral invertase, sucrose synthase and sucrose phosphate synthase levels were comparable. Those results and linkage analysis indicate that Rs is a candidate locus for carrot vacuolar acid soluble invertase isozyme II. Although the 2.5 kb insert does not occur in the Rs wild-type acid-soluble invertase isozyme II allele, it does occur elsewhere in the genome of Rs/Rs plants. PMID- 14756314 TI - Conservation and divergence of ASK1 and ASK2 gene functions during male meiosis in Arabidopsis thaliana. AB - Selective proteolysis of regulatory proteins mediated by the ubiquitin pathway is an important mechanism for controlling many biological events. The SCF (Skpl Cullin-F-box protein) class of E3 ubiquitin ligases controls the ubiquitination of a wide variety of substrates, thereby mediating their degradation by the 26S proteasome. The Arabidopsis genome contains 21 genes encoding Skp1-like proteins that are named as ASKs (Arabidopsis Skp1-like). So far, only the ASK1 gene has been characterized genetically, and is known to be required for male meiosis, flower development, and auxin response. The ASK2 gene is most similar to ASK1 in terms of both the amino acid sequence and expression pattern. To compare ASK2 with ASK1 functionally in male meiosis, different transgenic lines over expressing ASK1 and ASK2 were tested for their ability to complement the male meiosis defect of the ask1-1 mutant. The genomic ASK1 rescued the ask1-1 mutant defects. The 35S::ASK1 transgene restored male fertility to the ask1-1 mutant, although the percentages of normal pollen grains and tetrads were reduced. 35S::ASK2 lines in the ask1-1 background exhibited partial fertility with even fewer normal pollen grains and tetrads than those of the 35S::ASK1 lines. Detailed analysis of chromosome behavior during male meiosis demonstrated that 35S::ASK1 and 35S::ASK2 lines had different fractions of pollen mother cells undergoing normal meiosis. Our results suggest that ASK2 partially substitutes for ASK1 if expressed at higher than normal levels. PMID- 14756315 TI - Bifunctional aldehyde/alcohol dehydrogenase (ADHE) in chlorophyte algal mitochondria. AB - Protein profiles of mitochondria isolated from the heterotrophic chlorophyte Polytomella sp. grown on ethanol at pH 6.0 and pH 3.7 were analyzed by Blue Native and denaturing polyacrylamide gel electrophoresis. Steady-state levels of oxidative phosphorylation complexes were influenced by external pH. Levels of an abundant, soluble, mitochondrial protein of 85 kDa and its corresponding mRNA increased at pH 6.0 relative to pH 3.7. N-terminal and internal sequencing of the 85 kDa mitochondrial protein together with the corresponding cDNA identified it as a bifunctional aldehyde/alcohol dehydrogenase (ADHE) with strong similarity to homologues from eubacteria and amitochondriate protists. A mitochondrial targeting sequence of 27 amino acids precedes the N-terminus of the mature mitochondrial protein. A gene encoding an ADHE homologue was also identified in the genome of Chlamydomonas reinhardtii, a photosynthetic relative of Polytomella. ADHE reveals a complex picture of sequence similarity among homologues. The lack of ADHE from archaebacteria indicates a eubacterial origin for the eukaryotic enzyme. Among eukaryotes, ADHE has hitherto been characteristic of anaerobes since it is essential to cytosolic energy metabolism of amitochondriate protists such as Giardia intestinalis and Entamoeba histolytica. Its abundance and expression pattern suggest an important role for ADHE in mitochondrial metabolism of Polytomella under the conditions studied. The current data are compatible with the view that Polytomella ADHE could be involved either in ethanol production or assimilation, or both, depending upon environmental conditions. Presence of ADHE in an oxygen-respiring algal mitochondrion and co-expression at ambient oxygen levels with respiratory chain components is unexpected with respect to the view that eukaryotes acquired ADHE genes specifically as an adaptation to an anaerobic lifestyle. PMID- 14756316 TI - Epoxide hydrolase: a mRNA induced by the fungal pathogen Alternaria alternata on rough lemon (Citrus jambhiri Lush). AB - An expression profile of genes induced by non-pathogenic Alternaria alternata on rough lemon leaves was obtained by sequencing 500 subtractive PCR clones generated from mRNA of leaves inoculated with the fungus after subtraction with that of non-inoculated leaves. About 6% of the cDNA sequences had homology to known putative defense-related genes including epoxide hydrolase. A full-length cDNA (951 bp) from rough lemon that encoded epoxy hydrolase was isolated by random amplification of cDNA ends (RACEs), based on sequence information from subtractive PCR, and designated as RlemEH. The product of this gene expressed with an in vitro translation system with Escherichia coli also had activity of a soluble type of epoxide hydrolase. The transcript of rough lemon RlemEH was not detected in flowers, fruits, stems or leaves, but was induced after inoculation of leaves with conidia of Alternaria alternata, wounding, or treatment with C6 volatiles, including trans-2-hexenol and cis-3-hexenol, and methyl jasmonate. The response of the epoxide hydrolase gene correlated well with the activation of defense mechanisms induced in plant-fungus interactions. PMID- 14756317 TI - Arabidopsis transportin1 is the nuclear import receptor for the circadian clock regulated RNA-binding protein AtGRP7. AB - We characterized the Arabidopsis orthologue of the human nuclear import receptor transportin1 (TRN1). Like the human receptor, Arabidopsis TRN1 recognizes nuclear import signals on proteins that are different from the classical basic nuclear localization signals. The M9 domain of human heterogeneous nuclear ribonucleoprotein A1 (hnRNP A1) is the prototype of such signals. We show that AtTRN1 binds to similar domains in hnRNP-like proteins from plants. AtTRN1 also interacts with human hnRNP A1 and with yeast Nab2p, two classical import cargo proteins of transportin in these organisms. Like all nuclear transport receptors of the importin-beta family, AtTRN1 binds to the regulatory GTPase Ran from Arabidopsis. We demonstrated that the amino terminus of AtTRN1 is necessary for this interaction. Recombinant AtTRN1 conferred nuclear import of fluorescently labelled BSA-M9 peptide conjugates in permeabilized HeLa cells, functionally replacing human TRN1 in these in vitro nuclear import assays. We identified three plant substrate proteins that interact with AtTRN1 and contain M9-like domains: a novel Arabidopsis hnRNP that shows high similarity to human hnRNP A1 and two small RNA-binding proteins from Arabidopsis, AtGRP7 and AtGRP8. Nuclear import activity of the M9-like domains of these plant proteins was demonstrated in vivo by their ability to confer partial nuclear re-localisation of a GFP fusion protein containing a nuclear export signal. In addition, fluorescently labelled AtGRP7 was specifically imported into nuclei of permeabilized HeLa cells by Arabidopsis AtTRN1 and human TRN1. These results suggest that the transportin mediated nuclear import pathway is highly conserved between man, yeast and plants. PMID- 14756319 TI - Cyclin D-knockout uncouples developmental progression from sugar availability. AB - Multicellular organisms need to modulate proliferation and differentiation in response to external conditions. An important role in these processes plays the mitogen-stimulated induction of cyclin D (cycD) gene expression. D-type cyclins have been identified as the crucial intracellular sensors for cell-cycle regulation in all eukaryotes. However, cycD deletions have been found to cause specific phenotypic alterations in animals but not yet in plants. An insertional mutation of a so far uncharacterized Arabidopsis cycD gene did not alter the plant phenotype. To gain new insights into CycD function of land plants, we generated targeted cycD gene knockouts in the moss Physcomitrella patens and observed a surprisingly limited disruption phenotype. While wild-type plants reacted to exogenous glucose sources with prolonged growth of juvenile stages and retarded differentiation, cycD knockouts exhibited developmental progression independent of sugar supply. On the other hand, growth rate, cell sizes or plant size were not affected. Thus, we conclude that Physcomitrella CycD might not be essential for cell-cycle regulation but is important for coupling the developmental progression to nutrient availability. PMID- 14756318 TI - Biochemical evidence linking a putative callose synthase gene with (1 --> 3)-beta D-glucan biosynthesis in barley. AB - A putative barley (1 --> 3)-beta-D-glucan synthase cDNA of 6.1 kb, which is homologous to the yeast FKS gene, was assembled from DNA fragments obtained through screening of barley cDNA and BAC libraries, and by PCR amplification. The corresponding gene, designated HvGSL1, is a member of a family of at least six genes in barley. Gene transcripts are detected at relatively high levels in early developing grain, florets, coleoptiles and roots, but not in leaves infected with a fungal pathogen. A (1 --> 3)-beta-D-glucan synthase has been purified more than 60-fold from barley suspension-cultured cells by detergent extraction, CaCl2 treatment, sucrose density gradient centrifugation and non-denaturing gel electrophoresis. The enzyme synthesizes (1 --> 3)-beta-D-glucan in vitro and is recognized by antibodies raised against a 17 kDa protein generated by heterologous expression of a fragment of the HvGSL1 cDNA. Furthermore, mass spectrometric analyses show that tryptic peptides produced by in-gel digestion of the active enzyme match peptides predicted from the gene sequence. Thus, the amino acid sequence predicted from the HvGSL1 gene has been linked with the actual amino acid sequence of an active (1 --> 3)-beta-D-glucan synthase fraction from barley. PMID- 14756320 TI - Isolation of a subfamily of genes for R2R3-MYB transcription factors showing up regulated expression under nitrogen nutrient-limited conditions. AB - Plant R2R3-MYB transcription factors are encoded by more than 100 copies of genes. In this study, we attempted to isolate some members of the R2R3-MYB superfamily involved in regulation of nitrogen fixation in legumes. A library of 300 recombinant plasmid clones containing the R2R3-MYB fragments of the superfamily was screened by differential hybridization to isolate R2R3-MYB genes whose expression was up-regulated under nitrogen nutrient-limited conditions. Two groups of clones were identified, each of which seemed to represent a gene responsive to nitrogen starvation. The entire coding regions for the genes were further isolated by PCR and were designated LjMYB101 and LjMYB102. By screening a genomic library of Lotus japonicus with a probe derived from LjMYB101, the third gene, LjMYB103, was isolated. In addition, a candidate for the soybean orthologue of LjMYB101 was isolated and designated GmMYB101. Sequence alignment of the genes with members of the plant R2R3-MYB superfamily showed that they all belonged to the subgroup 10 of the superfamily. The expression analysis of the genes showed that the organ-specific and nitrate-regulated expression profile of MYB101 was very similar to that of CHS in Lotus as well as in soybean, suggesting a possible role for MYB101 in regulation of flavonoid biosynthesis in response to nitrate starvation. On the other hand, an interesting relationship, in structure and function, was found between LjMYB101 and LjGln1, suggesting an alternative role for MYB101 in regulation of nitrogen metabolism. PMID- 14756322 TI - The morphogenesis of evolutionary developmental biology. AB - The early studies of evolutionary developmental biology (Evo-Devo) come from several sources. Tributaries flowing into Evo-Devo came from such disciplines as embryology, developmental genetics, evolutionary biology, ecology, paleontology, systematics, medical embryology and mathematical modeling. This essay will trace one of the major pathways, that from evolutionary embryology to Evo-Devo and it will show the interactions of this pathway with two other sources of Evo-Devo: ecological developmental biology and medical developmental biology. Together, these three fields are forming a more inclusive evolutionary developmental biology that is revitalizing and providing answers to old and important questions involving the formation of biodiversity on Earth. The phenotype of Evo-Devo is limited by internal constraints on what could be known given the methods and equipment of the time and it has been framed by external factors that include both academic and global politics. PMID- 14756321 TI - An Arabidopsis thaliana T-DNA mutagenized population (GABI-Kat) for flanking sequence tag-based reverse genetics. AB - The GABI-Kat population of T-DNA mutagenized Arabidopsis thaliana lines with sequence-characterized insertion sites is used extensively for efficient progress in plant functional genomics. Here we provide details about the establishment of the material, demonstrate the population's functionality and discuss results from quality control studies. T-DNA insertion mutants of the accession Columbia (Col 0) were created by Agrobacterium tumefaciens-mediated transformation. To allow selection of transformed plants under greenhouse conditions, a sulfadiazine resistance marker was employed. DNA from leaves of T1 plants was extracted and used as a template for PCR-based amplification of DNA fragments spanning insertion site borders. After sequencing, the data were placed in a flanking sequence tag (FST) database describing which mutant allele was present in which line. Analysis of the distribution of T-DNA insertions revealed a clear bias towards intergenic regions. Insertion sites appeared more frequent in regions in front of the ATG and after STOP codons of predicted genes. Segregation analysis for sulfadiazine resistance showed that 62% of the transformants contain an insertion at only one genetic locus. In quality control studies with gene specific primers in combination with T-DNA primers, 76% of insertions could be confirmed. Finally, the functionality of the GABI-Kat population was demonstrated by exemplary confirmation of several new transparent testa alleles, as well as a number of other mutants, which were identified on the basis of the FST data. PMID- 14756323 TI - The place of phylogeny and cladistics in Evo-Devo research. AB - Here we review the various uses to which phylogenetic trees may be put when analysing the evolution of organisms and of the genotypic and phenotypic characteristics of these organisms. We briefly discuss the cladistic method and its application in the inference of phylogenetictrees. Next we consider the uses to which phylogenetic trees can be put: in particular for determining the homology or otherwise of characters distributed on those trees and for estimating the likely characteristics of ancestral taxa. Finally we show the application of this information for deepening our understanding of the processes of evolution. All of these forms of inference are fundamental for comparative biology and of immediate importance to the practice of evolutionary developmental biology. PMID- 14756324 TI - Evo-Devo: evolutionary developmental mechanisms. AB - Evolutionary developmental biology (Evo-Devo) as a discipline is concerned, among other things, with discovering and understanding the role of changes in developmental mechanisms in the evolutionary origin of aspects of the phenotype. In a very real sense, Evo-Devo opens the black box between genotype and phenotype, or more properly, phenotypes as multiple life history stages arise in many organisms from a single genotype. Changes in the timing or positioning of an aspect of development in a descendant relative to an ancestor (heterochrony and heterotopy) were two evolutionary developmental mechanisms identified by Ernst Haeckel in the 1870s. Many more have since been identified, in large part because of our enhanced understanding of development and because new mechanisms emerge as development proceeds: the transfer from maternal to zygotic genomic control; cell to-cell interactions; cell differentiation and cell migration; embryonic inductions; functional interactions at the tissue and organ levels; growth. Within these emergent processes, gene networks and gene cascades (genetic modules) link the genotype with morphogenetic units (cellular modules, namely germ layers, embryonic fields or cellular condensations), while epigenetic processes such as embryonic inductions, tissue interactions and functional integration, link morphogenetic units to the phenotype. Evolutionary developmental mechanisms also include interactions between individuals of the same species, individuals of different species, and species and their biotic and/or abiotic environment. Such interactions link ecological communities. Importantly, there is little to distinguish the causality that underlies these interactions from that which underlies inductive interactions within embryos. PMID- 14756325 TI - 'De-evolution' of Drosophila toward a more generic mode of axis patterning. AB - The genetics of the establishment of the primary axes of the early embryo have been worked out in great detail Drosophila. However, evidence has accumulated that Drosophila employs a mode of patterning that is not shared with most insects. In particular, the use of the morphogenic gradient of the Bicoid homeoprotein appears to be a novel addition to the fly developmental toolkit. To better understand the ancestral mode of patterning that is probably more widely used by insects, several groups have used Evo-Devo approaches as well as sophisticated genetic manipulations of Drosophila to achieve some form of 'de evolution' of this derived insect. Genetic manipulations of the beetle Tribolium and the wasp Nasonia have validated most of these results. PMID- 14756326 TI - The Cambrian "explosion" of metazoans and molecular biology: would Darwin be satisfied? AB - The origins of metazoan bodyplans and the extent to which they are coincident with the Cambrian "explosion" are both areas of continuing debate. The fossil record has a unique advantage in terms of historical perspective, but remains highly contentious on account of the often controversial interpretations of particular groups (e.g. halkieriids, vetulicolians) and the heavy reliance on "windows" of exceptional preservation (e.g. Chengjiang, Burgess Shale). Molecular and developmental biology offer other unique insights, but may be problematic in terms of conflicting phylogenetic signals and questions revolving around gene co option, evolution of developmental systems and even convergence. Such topics, far from frustrating the enterprise, actually widen our understanding of the nature of the evolutionary process with the exciting promise of the discovery of more general principles. PMID- 14756327 TI - Morphological and developmental macroevolution: a paleontological perspective. AB - Evidence of the morphological evolution of metazoans has been preserved, in varying degrees of completeness, in the fossil record of the last 600 million years. Although extinction has been incessant at lower taxonomic levels, genomic comparisons among surviving members of higher taxa suggest that much of the developmental systems that pattern their bodyplans has been conserved from early in their history. Comparisons between the origin of morphological disparity in the record and patterns of genomic disparity among living taxa promise to be interesting. For example, Hox cluster composition varies among major taxa, and the fossil record suggests that many of the changes in Hox clusters may have been associated with late Neoproterozoic evolution among minute benthic vermiform clades, from which crown bilaterian phyla arose just before or during the Cambrian explosion. Study of genomic differences among crown classes and orders whosetiming and mode of origin can be inferred from morphological data inthefossil record should throw further light on the timing and mode of origin of genomic disparities. PMID- 14756328 TI - The origins of axial patterning in the metazoa: how old is bilateral symmetry? AB - Bilateral symmetry is a hallmark of the Bilateria. It is achieved by the intersection of two orthogonal axes of polarity: the anterior-posterior (A-P) axis and the dorsal-ventral (D-V) axis. It is widely thought that bilateral symmetry evolved in the common ancestor of the Bilateria. However, it has long been known that members of the phylum Cnidaria, an outgroup to the Bilateria, also exhibit bilateral symmetry. Recent studies have examined the developmental expression of axial patterning genes in members of the phylum Cnidaria. Hox genes play a conserved role in patterning the A-P axis of bilaterians. Hox genes are expressed in staggered axial domains along the oral-aboral axis of cnidarians, suggesting that Hox patterning of the primary body axis was already present in the cnidarian-bilaterian ancestor. Dpp plays a conserved role patterning the D-V axis of bilaterians. Asymmetric expression of dpp about the directive axis of cnidarians implies that this patterning system is similarly ancient. Taken together, these result imply that bilateral symmetry had already evolved before the Cnidaria diverged from the Bilateria. PMID- 14756329 TI - Origin and evolution of endoderm and mesoderm. AB - Germ layers are defined as cell layers that arise during early animal development, mostly during gastrulation, and that give rise to all tissues and organs in adults. The evolutionary origin of the inner germ layers, endoderm and mesoderm, and their relationship have been a matter of debate for decades. In this review we summarize the major modes of endoderm and mesoderm formation found in Metazoa and possible evolutionary scenarios to reconstruct the ancestral state. In the second part, we address the question whether endoderm as well as mesoderm are homologous among Bilateria. In this regard, we propose that the comparative analysis of some crucial transcription factors involved in the early specification and differentiation of these germ layers might provide cues for the level of homology. We focus on four classes of genes: the Zn-finger gene GATA 4 6, the bHLH gene twist, the Kruppel-like Zn-finger gene snail and the T-box gene brachyury. The role of each of these genes in mesendoderm formation is summarized and we propose that the specific function of each of these genes in endoderm and mesoderm formation evolved from the regulation of basic cellular features, such as cell adhesion, cell motility, cytoskeleton and cell cycle. PMID- 14756330 TI - Origins and plasticity of neural crest cells and their roles in jaw and craniofacial evolution. AB - The vertebrate head is a complex assemblage of cranial specializations, including the central and peripheral nervous systems, viscero- and neurocranium, musculature and connective tissue. The primary differences that exist between vertebrates and other chordates relate to their craniofacial organization. Therefore, evolution of the head is considered fundamental to the origins of vertebrates (Gans and Northcutt, 1983). The transition from invertebrate to vertebrate chordates was a multistep process, involving the formation and patterning of many new cell types and tissues. The evolution of early vertebrates, such as jawless fish, was accompanied by the emergence of a specialized set of cells, called neural crest cells which have long held a fascination for developmental and evolutionary biologists due to their considerable influence on the complex development of the vertebrate head. Although it has been classically thought that protochordates lacked neural crest counterparts, the recent identification and characterization of amphioxus and ascidian genes homologous to those involved in vertebrate neural crest development challenges this idea. Instead it suggests thatthe neural crest may not be a novel vertebrate cell population, but could have in fact originated from the protochordate dorsal midline epidermis. Consequently, the evolution of the neural crest cells could be reconsidered in terms of the acquisition of new cell properties such as delamination-migration and also multipotency which were key innovations that contributed to craniofacial development. In this review we discuss recent findings concerning the inductive origins of neural crest cells, as well as new insights into the mechanisms patterning this cell population and the subsequent influence this has had on craniofacial evolution. PMID- 14756331 TI - The origin and evolution of the nervous system. AB - The nervous systems of animals as diverse as flies and mice share many conserved features, suggesting that such features were already present in their last common ancestor. As our knowledge of neural development increases, so does the list of conserved features, pointing to the existence of a highly sophisticated, single species as the origin of most extant nervous systems. Possible reasons for this unexpected monophyly are discussed, leading to the conclusion that the appearance of very different life forms, lifestyles and habitats requires the previous attainment of a neural circuitry that is sufficiently robust to cope with large changes without losing its overall coherence. PMID- 14756332 TI - Evolution of eyes and photoreceptor cell types. AB - The evolution of the eye is a matter of debate ever since Darwin's Origin of Species. While morphological comparisons of eye anatomy and photoreceptor cell types led to the view that animal eyes evolved multiple times independently, the molecular conservation of the pax6 eye-specifying cascade has indicated the contrary - that animal eyes evolved from a common, simple precursor, the proto eye. Morphological and molecular comparative approaches are combined here in a novel Evo-Devo approach, the molecular comparison of cell types ("comparative molecular cell biology"). In the eye, the various types of photoreceptor cells, as well as pigment and lens cells, each require distinct combinations of specifying transcription factors that control their particular differentiation programmes, such as opsin expression in photoreceptors, specific neurotransmitter metabolism, or axonal outgrowth. Comparing the molecular combinatorial codes of cell types of animal extant eyes, their evolutionary histories can be reconstructed. This is exemplified here on the evolution of ciliary and rhabdomeric photoreceptor cells in bilaterian eyes and on the evolution of cell type diversity in the vertebrate retina. I propose that the retinal ganglion, amacrine and horizontal cells are evolutionary sister cell types that evolved from a common rhabdomeric photoreceptor cell precursor. PMID- 14756333 TI - The origin and evolution of appendages. AB - Current awareness of gene expression patterns and developmental mechanisms involved in the outgrowth and patterning of animal appendages contributes to our understanding of the origin and evolution of these body parts. Nevertheless, this vision needs to be complemented by a new adequate comparative framework, in the context of a factorial notion of homology. It may even be profitable to categorize as appendages also gut diverticula, body ingrowths and 'virtual appendages' such as the eye spots on butterfly wings. Another unwarranted framework is the Cartesian co-ordinate system onto which the appendages are currently described and where it is supposed that one patterning system exists for each separate Cartesian axis. It may be justified, instead, to look for correspondences between the appendages and the main body axis of the same animal, as the latter might be the source of the growth and patterning mechanisms which gave rise to the former. This hypothesis of axis paramorphisms is contrasted with the current hypothesis of gene co-option. Recapitulationism is a common fault in current Evo-Devo perspectives concerning the origin of the appendages, in that the evolutionary origin of appendages is often expected to be the same as one of the key mechanisms involved in the ontogenetic inception of appendage formation. This unwarranted perspective is also evident in the current debate on the nature of the default arthropod appendage. Most likely, a default arthropod appendage never did exist, as the first appendages probably developed along the trunk of an animal already patterned extensively along the antero-posterior body axis. PMID- 14756334 TI - Segmentation: mono- or polyphyletic? AB - Understanding the evolutionary origins of segmented body plans in the metazoa has been a long-standing fascination for scientists. Competing hypotheses explaining the presence of distinct segmented taxa range from the suggestion that all segmentation in the metazoa is homologous to the proposal that segmentation arose independently many times, even within an individual clade or species. A major new source of information regarding the extent of homology vs. homoplasy of segmentation in recent years has been an examination of the extent to which molecular mechanisms underlying the segmentation process are conserved, the rationale being that a shared history will be apparent by the presence of common molecular components of a developmental program that give rise to a segmented body plan. There has been substantial progress recently in understanding the molecular mechanisms underlying the segmentation process in many groups, specifically within the three overtly segmented phyla: Annelida, Arthropoda and Chordata. This review will discuss what we currently know about the segmentation process in each group and how our understanding of the development of segmented structures in distinct taxa have influenced the hypotheses explaining the presence of a segmented body plan in the metazoa. PMID- 14756336 TI - Evolution of the Hox/ParaHox gene clusters. AB - The Hox gene cluster is a guiding force within the field of Evolutionary Developmental Biology. In large part our understanding of this gene cluster comes from only a few model organisms in developmental biology. The situation is gradually changing. A comparative review of the organisation of the Hox and ParaHox gene clusters and, in particular, instances of cluster disintegration, leads us to the view that the phenomenon of Temporal Colinearity is the major constraining force in maintaining these gene clusters over such long evolutionary timespans. PMID- 14756335 TI - Vertebrate somitogenesis: a novel paradigm for animal segmentation? AB - In vertebrates, the primary segmented tissue of the body axis is the paraxial mesoderm, which lies bilaterally to the axial organs, neural tube and notochord. The segmental pattern of the paraxial mesoderm is established during embryogenesis through the production of the somites which are transient embryonic segments giving rise to the vertebrae, the skeletal muscles and the dorsal dermis. Somitogenesis can be subdivided into three major phases (see Fig. 1). First a growth phase during which new paraxial mesoderm cells are produced by a growth zone (epiblast and blastopore margin or primitive streak and later on tail bud) and become organized as two rods of mesenchymal tissue,forming the presomitic mesoderm. Second a patterning phase occuring in the PSM, during which the segmental pattern is established at the molecular level. Third, the somitic boundaries are formed during the morphological segmentation phase. In all vertebrates, all cells of the paraxial mesoderm, during their maturation in the PSM, go successively through these three phases, which are tightly regulated at the spatio-temporal level. The first phase of paraxial mesoderm production falls out of the scope of this review, as it essentially pertains to the gastrulation process. Here, I essentially discuss the segmental patterning phase in vertebrates. Recent data suggest that establishment of the segmental pattern relies on a clock and wavefront mechanism which has been conserved in vertebrates. Furthermore, conservation of this system could extend to invertebrates, suggesting that the clock and wavefront is an ancestral mechanism. PMID- 14756337 TI - Time's arrow: heterochrony and the evolution of development. AB - The concept of heterochrony, which denotes a change in the relative timing of developmental events and processes in evolution, has accompanied attempts to link evolution and development for well over a century. During this time the definition of heterochrony and the application of the concept have varied and by the late 1990's, many questioned the usefulness of the concept. However, in the past decade studies of heterochrony have been revitalized by a new focus on developmental sequence, an examination of heterochrony in explicit phylogenetic contexts and increasing tendencies to examine the heterochrony of many kinds of events, including cellular, molecular and genetic events. Examples of such studies are reviewed in this paper and it is argued that this new application of heterochrony provides an extraordinarily rich opportunity for understanding the developmental basis of evolutionary change. PMID- 14756338 TI - Who came first--larvae or adults? origins of bilaterian metazoan larvae. AB - There is a classic controversy in zoology over whether the common ancestor of living bilaterian phyla was a benthic animal with a bilaterian body plan, or was a pelagic larva-like animal similar to what we see today in the primary larvae of indirect-developing bilaterians. We examine the current larva-like adult hypothesis, and present an alternate model for the evolution of complex life histories by intercalation of larval features into the ontogeny of an ancestral direct-developing bilaterian. This gradual accumulation of larval features results in a developmental regulatory program that produces a larva distinct in body plan from the adult. The evolution of a rapid and complete metamorphosis is made possible by the convergent evolution of set aside cells in the final stages of the emergence of indirect developing larval forms. Although convergences abound either hypothesis for the evolution of developmental pathways and life histories, the bilaterian first hypothesis is consistent with all stages of evolution of a complex life history being selectively advantageous, with the rapid evolution of larval forms, and with the frequent co-option of genes from the adult phase of the life cycle prevalent in the evolution of embryos and larvae. PMID- 14756339 TI - Polyembryony in parasitic wasps: evolution of a novel mode of development. AB - Major developmental innovations have been associated with adaptive radiations that have allowed particular groups of organisms to occupy empty ecospace. Well known developmental novelties associated with the conquest of new habitats include the evolution of the tetrapode limb, allowing the radiation of vertebrates into a terrestrial habitat, and formation of insect wings that permitted their dispersal into the air. However, an understanding of the evolutionary forces and molecular mechanisms behind developmental novelties still remains tenuous. A little-studied adaptive radiation in insects from the developmental perspective is the evolution of parasitism. The parasitic lifestyle has allowed parasitic insects to occupy a novel ecological niche where they have evolved a plethora of life history strategies and modes of embryogenesis, developing on or within the body of the host. One of the most striking adaptations to development within the body of the host includes polyembryonic development, where certain wasps form clonally up to 2000 embryos from a single egg. Taking advantage of well-established insect phylogeny, techniques developed in a model insect, the fruit fly, and a wealth of knowledge in comparative insect embryology, we are starting to tease apart the evolutionary events that have led to this novel mode of development in insects. PMID- 14756340 TI - Evolution of cis-regulation of the proneural genes. AB - The current state of knowledge concerning cis-regulatory sequences of the proneural genes of vertebrates and Drosophila is discussed. Many proneural genes have a complex modular arrangement of discrete enhancer elements. One unusual feature of these genes is that many distant enhancer elements, regulating expression in specific spatial locations, require input from previously synthesized protein from the proneural gene itself, in addition to other transcriptional activators. This is distinct from the auto-regulation, via E boxes in the promoter, that takes place in neural precursors. The selection of neural precursors from a field of cells expressing a proneural gene, is mediated by Notch signalling and requires up-regulation of proneural gene expression in the precursor concurrently with down-regulation in the surrounding cells. Although the way in which a single cell is selected remains unclear, a number of feedback loops have been uncovered that reinforce the choice. These are briefly surveyed. A specific regulatory element, the Sensory Organ Precursor element, that mediates selection of the precursors of the large sensory bristles, has been described in Drosophila. We report the conservation of this sequence in Calliphora vicina, a higher fly. In contrast, no such sequence is seen in the achaete-scute complex of Anopheles gambiae, a basal Dipteran species. We suggest that this enhancer may have arisen during the evolution of the cyclorraphous flies and present a hypothesis for its possible function. PMID- 14756341 TI - Developmental basis for vein pattern variations in insect wings. AB - The venation patterns characteristics of different insect orders and of families belonging to the same order possess enormous variation in vein number, position and differentiation. Although the developmental basis of changes in vein patterns during evolution is entirely unknown, the identification of the genes and developmental processes involved in Drosophila vein pattern formation facilitates the elaboration of construction rules. It is thus possible to identify the likely changes which may constitute a source of pattern variation during evolution. In this review, we discuss how actual patterns of venation could be accounted for by modifications in different Pterygota of a common set of developmental operations. We argue that the individual specification of each vein and the modular structure of the regulatory regions of the key genes identified in Drosophila offer candidate entry points for pattern modifications affecting individual veins or interveins independently. Assuming a general conservation of the processes involved in different species, the transitions between different patterns may require few changes in the regulatory gene networks involved. PMID- 14756343 TI - Transcriptional regulation and the evolution of development. AB - A growing body of evidence suggests that changes in transcriptional regulation form an important part of the genetic basis for the evolution of development. At a microevolutionary level, all the necessary conditions are present: populations harbor abundant genetic variation for differences in transcription profiles, a substantial fraction of these variants can influence organismal phenotype, and some variants have fitness consequences and are subject to natural selection. At a macroevolutionary level, the evidence is less direct but strongly suggestive: specific differences in anatomy and gene expression are often correlated, while comparisons of transcription profiles among distantly related taxa point to extensive evolutionary changes in regulatory gene networks. Understanding how transcriptional regulatory systems evolve, and what contributions these changes have made to the evolution of phenotype, represents a major challenge for Evo Devo. PMID- 14756342 TI - Evolution of cis-regulatory regions versus codifying regions. AB - Efforts to understand the genetic basis of evolutionary change have concentrated on proteins and their encoding DNA sequences. These studies have brought to light patterns and processes at the nucleotide level, yet the complex functional relationships between genetic variants and phenotypes remain poorly known. The realization that even a complete description of proteins and the effects of their activity will not suffice to understand the conditions under which they are time- and tissue-specifically expressed or repressed during development has refocused attention on cis-regulatory regions. In particular, promoter sequences are thought to hold the key for understanding the evolution of phenotypic differences between species. This is because of their complex organization into independent modules such that, unlike coding sequences in which mutations affect protein function every time the protein is expressed, mutations in cis-regulatory sequences may have minor or no pleiotropic effects. Complex information-encoding makes cis-regulatory regions poorly amenable to comparative methods designed for coding sequences. Some general conclusions are emerging as to how genetic variation is distributed across regulatory networks and the processes modulating the structure of this variation. We bring into this emerging scenario several recent findings pointing to different ways in which spliceosomal introns, pseudogenes and patterns of point mutation can be active players for the evolution of novel transcriptional profiles. PMID- 14756344 TI - Adaptive walks in a gene network model of morphogenesis: insights into the Cambrian explosion. AB - The emergence of complex patterns of organization close to the Cambrian boundary is known to have happened over a (geologically) short period of time. It involved the rapid diversification of body plans and stands as one of the major transitions in evolution. How it took place is a controversial issue. Here we explore this problem by considering a simple model of pattern formation in multicellular organisms. By modeling gene network-based morphogenesis and its evolution through adaptive walks, we explore the question of how combinatorial explosions might have been actually involved in the Cambrian event. Here we show that a small amount of genetic complexity including both gene regulation and cell cell signaling allows one to generate an extraordinary repertoire of stable spatial patterns of gene expression compatible with observed anteroposterior patterns in early development of metazoans. The consequences for the understanding of the tempo and mode of the Cambrian event are outlined. PMID- 14756345 TI - Developmental gene network analysis. AB - The developmental process is controlled by the information processing functions executed by the cis-elements that regulate the expression of the participating genes. A model of the network of cis-regulatory interactions that underlies the specification of the endomesoderm of the sea urchin embryo is analyzed here. Although not all the relevant interactions have yet been uncovered, the model shows how the information processing functions executed by the cis-regulatory elements involved can control essential functions of the specification process, such as transforming the localization of maternal factors into a domain-specific program of gene expression; refining the specification pattern; and stabilizing states of specification. The analysis suggests that the progressivity of the developmental process is also controlled by the cis-regulatory interactions unraveled by the network model. Given that evolution occurs by changing the program for development of the body plan, we illustrate the potential of developmental gene network analysis in understanding the process by which morphological features are maintained and diversify. Comparison of the network of cis-regulatory interactions with a portion of that underlying the specification of the endomesoderm of the starfish illustrates how the similarities and differences provide insights into how the programs for development work and how they evolve. PMID- 14756346 TI - Evo-Devo: the long and winding road. AB - Evolutionary developmental biology (Evo-Devo) aims to unveil how developmental processes and mechanisms become modified during evolution and how from these changes the past and present biodiversity arose. The first wave of Evo-Devo identified a conserved set of toolkits common to most metazoans. The present second wave has changed gear and aims to identify how genes and modules were used differently through evolution to build the past and present morphological diversity. The burgeoning third wave is introducing experimental testing of predictions drawn from the first and second waves. Here we review some of the hottest topics, contributions and insights of present Evo-Devo related to basic concepts and paradigms of evolutionary research. Future directions of Evo-Devo are also highlighted; in other words, Quo Vadis, Evo-Devo? PMID- 14756347 TI - History of ultrasound in cardiology. PMID- 14756348 TI - Effects of hypertension and antihypertensive treatment on retrobulbar circulation detected on Doppler sonography. AB - OBJECTIVE: To study the effect of antihypertensive drugs in essential hypertension and in the central retinal, posterior ciliary, and ophthalmic arteries by using Doppler sonography. METHODS: Thirty patients with essential hypertension and 32 control subjects were enrolled in the study. The pulsatility and resistive indices were evaluated in the central retinal, posterior ciliary, and ophthalmic arteries before and 2 months after treatment with antihypertensive drugs as well as in the control group. The pulsatility and resistive index values for each artery in each group were compared statistically. RESULTS: There were significant differences in the retrobulbar pulsatility and resistive index values in each artery among the patients with initially diagnosed hypertension and the control group (P < .05). There were significant reductions in the resistive and pulsatility index values of the posterior ciliary and ophthalmic arteries after treatment (P < .05). For the ophthalmic artery, posttreatment pulsatility and resistive index values did not reach the level of flow measured in the control subjects. CONCLUSIONS: The alteration of Doppler parameters of medication-free patients with hypertension may result from a peripheral vasospasm in the retrobulbar circulation, and the improvement in the Doppler parameters with oral antihypertensive drugs may indicate the importance of early diagnosis in ameliorating hypertension-induced retrobulbar circulation changes. PMID- 14756349 TI - Detection and characterization of perianal inflammatory disease: accuracy of transperineal combined gray scale and color Doppler sonography. AB - OBJECTIVE: To evaluate the accuracy of transperineal gray scale and color Doppler sonography for the detection and characterization of perianal inflammatory disease with surgical correlation. METHODS: Eighty-seven patients with suspected perianal inflammatory disorders underwent transperineal gray scale and color Doppler sonography with a linear 4- to 7-MHz transducer that was used to scan the entire perianal region for the detection of suspected inflammatory disorders. Each detected inflammatory disorder was evaluated to determine its morphologic characteristics and extent. Color Doppler sonography was applied to assess the presence of increased vascularity in the perianal region. In comparison with surgical findings, the diagnostic performance of transperineal sonography was assessed by means of receiver operating characteristic analysis for lesion detection and the Spearman rho test for lesion characterization. Logistic regression analysis was used to assess whether increased perineal vascularity was a predictive factor of perianal inflammatory disease. RESULTS: Seventy-seven perianal inflammatory disorders were confirmed in 62 patients. Gray scale sonography achieved a significantly good performance in the detection (area under the curve = 0.86; P < .001) and characterization (r = 0.65; P < .001) of perianal inflammatory disease. For the detection of perianal fistulas and abscesses, sensitivity was 100% for both, and specificity was 100% and 94%, respectively. With the use of color Doppler sonography, the diagnostic confidence increased slightly (area under the curve = 0.89) but significantly (P = .002). Logistic regression analysis identified hypervascularity at the periphery of a perianal lesion as a significant independent predictor of an inflammatory disease. CONCLUSIONS: Combined gray scale and color Doppler sonography enables a high detectability rate and comprehensive characterization of perianal abscesses and fistulas. PMID- 14756350 TI - Endothelial cell injury and platelet aggregation induced by contrast ultrasonography in the rat hepatic sinusoid. AB - OBJECTIVE: To determine whether contrast ultrasonography can affect the sinusoidal cells and platelets of the liver by using ultrastructural analysis in vivo. METHODS: Fifteen Wistar rats were placed into the following 5 groups of 3 rats each: 3 control groups comprising a sham operation group, a contrast agent injection-alone group, and an ultrasound exposure-alone group; and 2 contrast agent injection with ultrasound exposure groups, split according to excision time. After a dose of an echo contrast agent (100 mg/kg of body weight) was administered through the femoral vein, the rats that received injections were subjected to ultrasound for the first minute, no ultrasound for the next 4 minutes, and then ultrasound sweep scanning for 10 seconds. The rats were perfused via the heart with cold physiologic saline containing 2% paraformaldehyde and 2.5% glutaraldehyde solution buffered with 0.1-mol/L phosphate. The livers of the rats in 4 of the groups were excised immediately. The livers of the rats in 1 of the 2 contrast agent with ultrasound exposure groups were excised by the same procedure 5 hours after they received the injections. All specimens were studied with light and electron microscopy. RESULTS: Platelet aggregation and injury to endothelial cells were more severe in the contrast agent injection and ultrasound exposure groups than in the other groups. CONCLUSIONS: Contrast ultrasonography can cause platelet aggregation and endothelial cell damage in the rat hepatic sinusoid. PMID- 14756351 TI - Quantification of tumor vascularity with contrast-enhanced sonography: correlation with magnetic resonance imaging and fluorodeoxyglucose autoradiography in an implanted tumor. AB - OBJECTIVE: To correlate the quantitated tumor vascularity of implanted murine tumors as depicted by contrast-enhanced sonography with estimates made with magnetic resonance imaging and with estimates of the percentage of viable (metabolically active) tumor as depicted by fluorodeoxyglucose autoradiography. METHODS: Implanted tumors in 10 mice were imaged with contrast-enhanced sonography, magnetic resonance imaging, and fluorodeoxyglucose autoradiography. Tumor vascularity was estimated with each modality and compared with the percentage of viable tumor. RESULTS: Quantitated estimates of tumor vascularity with contrast-enhanced sonography closely correlated (r = 0.95) with estimates made by magnetic resonance imaging and with the percentage of viable tumor (r = 0.93) as depicted by fluorodeoxyglucose autoradiography. CONCLUSIONS: Contrast enhanced sonography accurately depicts tumor vascularity in these implanted tumors. Tumor vascularity correlated with the amount of metabolically active tumor. PMID- 14756352 TI - Outcomes of pregnancies with sonographically detected nuchal cords remote from delivery. AB - OBJECTIVE: To evaluate the outcomes of pregnancies with sonographically detected nuchal cords remote from delivery during the second and third trimesters of gestation. METHODS: A retrospective study of patients with and without nuchal cords detected by sonography during the second and third trimesters of gestation (17-36 weeks) was conducted. Variables included maternal age, parity, gestational age at sonography and at delivery, method of delivery, intrapartum fetal heart abnormalities, meconium-stained amniotic fluid, birth weight, 5-minute Apgar score, and neonatal intensive care unit admissions. RESULTS: A total of 233 patients were included in this study, of which 118 had sonographically detected nuchal cords identified in our department during the study period. There were no statistically significant differences between patients with sonographically detected nuchal cords and control patients with respect to maternal age, estimated gestational age at sonography, method of delivery, meconium-stained fluid, birth weight, intrapartum fetal heart abnormalities, 5-minute Apgar scores of less than 7, and neonatal intensive care unit admissions. Patients with nuchal cords detected by sonography were more likely to give birth before 37 weeks' gestation than control patients (21% versus 11%; P = .040). CONCLUSIONS: A sonographically detected nuchal cord is not associated with important perinatal complications. PMID- 14756353 TI - Sonographic markers of hemoglobin Bart disease at midpregnancy. AB - OBJECTIVE: To evaluate the efficacy of various sonographic markers at midpregnancy in predicting fetal hemoglobin Bart disease. METHODS: Four hundred eighty-eight pregnancies at risk of having fetuses with hemoglobin Bart disease were recruited for prenatal diagnosis with cordocentesis at 18 to 21 gestational weeks. Before cordocentesis, the sonographic markers, including cardiothoracic ratio, placental thickness, pericardial effusion, pleural effusion, ascites, subcutaneous edema, cord edema, dilated umbilical vein, and amniotic fluid index, were assessed and recorded. The definite fetal diagnosis was based on blood analysis. The efficacy of each sonographic marker in predicting hemoglobin Bart disease was evaluated by sensitivity and specificity. RESULTS: Among 488 pregnancies undergoing prenatal diagnosis, 100 fetuses were proved to be affected by hemoglobin Bart disease. The cardiothoracic ratio gave the highest sensitivity, 95.0%, with specificity of 96.1%, followed by placental thickness. Signs of hydrops fetalis were observed in 33.0% of cases; they did not increase the sensitivity of the cardiothoracic ratio but strongly reinforced the diagnosis when they appeared. CONCLUSIONS: At midpregnancy, sonographic markers can effectively differentiate normal pregnancies from those with fetal hemoglobin Bart disease. Among couples at risk with no sonographic markers, the risk of having an affected child is nearly eliminated. The most sensitive marker was the cardiothoracic ratio, followed by placental thickness. PMID- 14756354 TI - Sonographic comparison of the tubal ring of ectopic pregnancy with the corpus luteum. AB - OBJECTIVE: Pregnant patients without a sonographically visible intrauterine pregnancy and with a thick-walled cystic adnexal structure present a dilemma. This study compared the utility of various sonographic features in differentiating between the tubal ring of ectopic pregnancy and the corpus luteum. METHODS: Retrospective review of first-trimester transvaginal sonograms revealed a cystic adnexal structure in 79 women. Each structure was evaluated for 6 specific sonographic characteristics: echogenicity of its wall compared with that of the ovary and endometrium, wall thickness in 2 planes, color Doppler flow distribution and percentage of wall circumference, and internal texture. RESULTS: Forty-one (52%) of the 79 women had ectopic pregnancies, and 38 (48%) had corpora lutea. Eleven (32%) of 35 ectopic walls were more echogenic than the endometrium, compared with none of the corpora lutea. A cyst wall less echogenic than the endometrium was more likely in corpora lutea (84% versus 31%; P < .0001). More than twice as many ectopic rinds were more echogenic than ovarian tissue compared with corpora lutea (76% versus 34%; P < .0001). The only predictive internal texture feature was a clear pattern, which was more common in the corpora lutea (P < .01, Fisher exact test). There was no significant difference in mural flow distribution or extent between the 2 groups. CONCLUSIONS: Ancillary sonographic signs to distinguish between an ectopic pregnancy and a corpus luteum include decreased wall echogenicity compared with the endometrium and an anechoic texture, which suggests a corpus luteum. PMID- 14756355 TI - Sonographic evaluation of isolated abnormal axillary lymph nodes identified on mammograms. AB - OBJECTIVE: To evaluate the role of sonography in evaluation of abnormal axillary lymph nodes identified in patients with otherwise negative or benign findings on mammography. METHODS: For 3 years 2 months, we retrospectively reviewed 30 consecutive cases that had undergone sonographic evaluation for abnormal axillary lymph nodes identified in patients whose mammograms had an American College of Radiology Breast Imaging Reporting and Data System final assessment of 1 or 2. Mammographic and sonographic features of the lymph nodes were analyzed and correlated with the histologic diagnosis in patients undergoing biopsy. Patients who did not undergo biopsy had clinical or imaging follow-up. RESULTS: Twenty of the 30 patients studied had an abnormal sonographic appearance. Biopsy was recommended in 17 of the 20 patients on the basis of an abnormal sonographic appearance. In the remaining 3 patients, there was an underlying cause for lymphadenopathy, and these patients underwent clinical and sonographic follow-up. Eighteen patients underwent biopsy, including 1 patient with normal findings on sonography. Ten of these patients had malignant histologic findings: 6 were metastatic adenocarcinoma; 1, poorly differentiated sarcoma, and 3, lymphoma. The remaining 8 patients had benign histologic findings. The nonbiopsy group had clinical and or imaging follow-up (mean, 17.6 months; range, 6-25 months). The sensitivity (true-positive/true-positive + false-negative) of sonography for assessment of suspected abnormal lymph nodes in the patients studied was 100% (10 of 10); specificity (true-negative/true-negative + false-positive), 50% (10 of 20); positive predictive value (true-positive/true-positive + false-positive) for malignancy based on the presence of 2 or more abnormal sonographic features, 50% (10 of 20); and negative predictive value, 100%. CONCLUSIONS: Sonography is useful in further characterization of isolated abnormal axillary lymph nodes identified on mammography. Sonographic evaluation helps improve the specificity of imaging evaluation in assessment of these lymph nodes. PMID- 14756356 TI - Posterior acoustic shadowing in benign breast lesions: sonographic-pathologic correlation. AB - OBJECTIVE: To show a variety of benign breast lesions that exhibit posterior acoustic shadowing on sonography. METHODS: The cases illustrate a variety of pathologic breast conditions that were collected at a referral breast center at a tertiary medical center. RESULTS: A variety of pathologic conditions are discussed, with pathologic-imaging correlation. CONCLUSIONS: Although posterior acoustic shadowing is a sonographic feature that is most commonly associated with mammary malignancies, this sonographic finding may also be seen with benign breast lesions. PMID- 14756357 TI - Variable breast conditions: comparison of conventional and real-time compound ultrasonography. AB - OBJECTIVES: To illustrate and compare the appearances of variable breast conditions by conventional and real-time spatial compound images. METHODS: Cases illustrative of a broad range of breast conditions were collected. Each image pair consisted of conventional and real-time compound images with a stationary probe to maintain an identical projection. RESULTS: The various breast conditions, including normal anatomic structures and abnormal lesions, were evaluated and compared by conventional and real-time compound images. The real time compound images revealed more realistic and clear images with reduced artifacts. CONCLUSIONS: Real-time compound images are superior to conventional images of normal and abnormal breast conditions. Real-time compound imaging is a good technique for evaluating the breast state. PMID- 14756358 TI - Saline infusion sonohysterography: technique, indications, and imaging findings. AB - OBJECTIVE: To review the technique, indications, and common imaging findings regarding saline infusion sonohysterography. METHODS: The literature on saline infusion sonohysterography was reviewed. Pertinent images from our institution are presented to illustrate common imaging findings. RESULTS: From the literature review, we summarize the various clinical scenarios in which saline infusion sonohysterography is useful and give examples from our clinical practice. CONCLUSIONS: Saline infusion sonohysterography is a useful procedure for evaluation of endometrial and subendometrial abnormalities. PMID- 14756359 TI - The onion skin sign: a specific sonographic marker of appendiceal mucocele. AB - OBJECTIVE: To evaluate the association of the onion skin sign as a sonographic marker for appendiceal mucocele. METHODS: The sonographic onion skin sign was considered specific for the preoperative diagnosis of appendiceal mucocele. Therefore, detection of this sign in a mass located in the right lower abdomen, unrelated to the female reproductive organs, indicated surgical intervention with a presumptive diagnosis of appendiceal mucocele. From 1998 through 2001, female patients who were found to have atypical cysts containing this sign underwent surgery. The cases were closely followed, and intraoperative findings and final histologic diagnoses were recorded. RESULTS: Appendiceal mucocele was the final diagnosis in all 7 patients in whom the onion skin sign was observed. One additional patient had an appendiceal mucocele with a sonographic picture of a clear tubular cystic structure. CONCLUSIONS: A sonographically layered cystic mass in the right lower quadrant of the abdomen in the presence of a normal ovary strongly suggests the diagnosis of appendiceal mucocele. Recognition of the sonographic onion skin sign in a cystic mass in the right lower quadrant may facilitate the accurate preoperative diagnosis of appendiceal mucocele. PMID- 14756360 TI - Congenital left ventricular aneurysm: prenatal sonographic diagnosis. PMID- 14756361 TI - Prenatal sonographic diagnosis of abdominal mesenteric lymphangioma. PMID- 14756362 TI - Contrast-enhanced sonography as guidance for transthoracic biopsy of a peripheral lung lesion with large necrotic areas. PMID- 14756363 TI - Gallbladder cryptosporidiosis in a patient with acquired immunodeficiency syndrome: sonographic evaluation of the disease's course. PMID- 14756364 TI - Gallbladder cancer appearing as a polyp within a polyp. PMID- 14756365 TI - High detail radiography and histology of the centrodistal tarsal joint of Icelandic horses age 6 months to 6 years. AB - REASONS FOR PERFORMING STUDY: Osteoarthrosis (OA) in the distal tarsal joints, bone spavin, is a well known condition which is common in Icelandic horses age 6 12 years. OBJECTIVES: To determine the nature, location and age of appearance of early radiographic and histological changes in the centrodistal tarsal joint (CD) of young Icelandic horses. METHODS: Slab sections from the CD of young Icelandic horses were examined by high detail radiography (age 6 months to 6 years, n = 111) and histology (age 6 months to 4 years, n = 82) to detect and describe the early changes indicative of OA. Horses younger than 5 years were unridden. RESULTS: Chondronecrotic lesions histologically similar to those described in the early pathogenesis of OA were seen in 33% of the joints, located both medially and laterally. Radiographic sclerosis of the subchondral bone was recorded in 60% of the specimens, most often medially. Medial location was not associated with chondronecrosis, but was strongly related to age. Sclerosis was an infrequent finding on the lateral side, and was probably secondary to chondronecrosis in the corresponding part of the joint. Small defects in the subchondral bone were considered to be the most specific radiographic sign of OA as they were strongly associated with chondronecrosis. CONCLUSIONS: The high prevalence of chondronecrosis in the young horses indicates an early onset and slow progression of the disease. The early appearance also shows that the initiation of the disease is unrelated to the use of horses for riding. As clinical manifestation of OA in the distal tarsal joints is most often described in mature or old horses, the first stages of the disease are not likely to result in clinical signs. Subchondral bone sclerosis did not appear to be a primary factor in the development of OA in the CD but was considered to reflect an uneven distribution of biomechanical forces within the joint. POTENTIAL RELEVANCE: The development of OA in the CD of young Icelandic horses seems to be due to poor conformation or joint architecture rather than trauma or overloading. These aetiological factors are likely to be of importance for OA in the distal tarsal joints in other breeds as well. The influence of hindlimb conformation and the architecture of the distal tarsal joints on the biomechanics of joints need to be investigated, preferably by locomotion analysis in young horses. PMID- 14756366 TI - Evaluation of decision criteria for detection of spinal cord compression based on cervical myelography in horses: 38 cases (1981-2001). AB - REASONS FOR PERFORMING STUDY: Different criteria have been described based on height reduction of the total myelographic contrast column and components of it as tests for compression of the spinal cord due to cervical stenotic myelopathy (CSM). Fifty percent height reduction of the dorsal myelographic column (DMC), <2 mm empiric height of the DMC and a 40% reduction of the ratio of stenosis calculated based on the height reduction of the entire dural diameter (DD) have been described as decision criteria for considering the test result positive. The reasons for selecting these decision criteria or their accuracies have rarely been reported. OBJECTIVES: To evaluate the accuracy of diagnostic criteria based on reduced height of the total myelographic column and components of it for diagnosing extradural spinal cord compression using different decision criteria, and make recommendations for consistent myelographic interpretation in horses suspected of having CSM. METHODS: Four measurements were obtained by 2 readers in a retrospective sample population of 38 horses in which both cervical myelography and histopathological examination of the cervical spinal cord were performed. The prevalence of CSM in the sample was 50%. At intervertebral sites, the minimum heights of the DD and DMC were measured. At intravertebral sites, the maximum heights of the entire DD and DMC were obtained. Percent height reductions of the DMC and DD were determined as the ratio of minimum intervertebral height to maximum intravertebral height within the next cranial vertebra. Histological examination was used as the gold standard for determining the actual site of spinal cord compression. Sensitivity and specificity for the diagnostic criteria were estimated at each site in neutral and flexed neck positions using several different decision criteria. CONCLUSIONS: At C6-C7, in neutral or flexed neck position and using 20% reduction of DD, the test was highly sensitive and specific for CSM. At other sites, reduced height of the myelographic column generally was not accurate for diagnosing extradural spinal cord compression. Using 20% reduction of DD in neutral position at the mid-cervical sites, the test had only low sensitivity and high specificity. Flexion of the neck appeared to increase detection of spinal cord compression in the mid-cervical region, but also substantially increased the frequency of false-positive diagnoses. POTENTIAL RELEVANCE: By using the reported sensitivity and specificity estimates, readers may decide on a decision criterion for diagnosis of extradural spinal cord compression due to CSM. However, in planning a surgical correction, it is difficult to define a decision criterion that combines acceptable sensitivity and specificity, especially at the mid-cervical sites. PMID- 14756367 TI - Measurement of respiratory function by impulse oscillometry in horses. AB - REASONS FOR PERFORMING STUDY: Due to technical implementations and lack of sensitivity, pulmonary function tests are seldom used in clinical practice. Impulse oscillometry (IOS) could represent an alternative method. OBJECTIVES: To define feasibility, methodology and repeatability of IOS, a forced oscillation technique that measures respiratory resistance (Rrs) and reactance (Xrs) from 5 to 35 Hz during spontaneous breathing, in horses. METHODS: Using 38 healthy horses, Rrs and Xrs reference values were defined and influence of individual biometrical parameters was investigated. In addition, IOS measurements of 6 horses showing clinical signs of heaves were compared to those of 6 healthy horses. RESULTS: Airtightness and minimal dead space in the facemask were prerequisites to IOS testing and standardisation of head position was necessary to avoid variations in Rrs due to modified upper airway geometry. In both healthy and diseased animals, measurements were repeatable. In standard-type breeds, the influence of the horse's size on IOS parameters was negligible. An increase in R5Hz greater than 0.10 kPa/l/sec and R5Hz>R10Hz, combined with negative values of Xrs between 5 and 20 Hz, was indicative of heaves crisis. CONCLUSIONS: IOS is a quick, minimally invasive and informative method for pulmonary function testing in healthy and diseased horses. POTENTIAL RELEVANCE: IOS is a promising method for routine and/or field respiratory clinical testing in the equine species. PMID- 14756368 TI - Total and partial ovariohysterectomy in seven mares. AB - REASONS FOR PERFORMING STUDY: Ovariohysterectomy appears to have a low mortality rate in mares, but the procedure needs to be reviewed because of the high risk of life-threatening complications. HYPOTHESIS: That ovariohysterectomy can be effective treatment for a variety of uterine diseases in mares and carries a good prognosis. METHODS: Diagnosis, clinical data, surgical technique, post operative care, complications and outcome were recorded from medical records of 7 mares that underwent total (6) and partial (1) ovariohysterectomy at the University of Illinois from 1994 to 2001. RESULTS: The indications for ovariohysterectomy were chronic pyometra (4 mares), chronic uterine torsion (n = 2) and chronic intramural haematoma (n = 1). Surgical exposure was difficult but was improved by traction on stay sutures and right-angled clamps. In some cases, application of the TA-90 autosuture instrument as a right-angled clamp to the caudal part of the uterus improved access to the uterine stump. The most common post operative complications were decreased faecal output, decreased intestinal sounds (4 mares) and mild abdominal pain (2). Two mares had mild to moderate incisional infections. Other previously reported complications, such as haemorrhage, septic peritonitis, uterine stump infection or necrosis, and diarrhoea, did not occur. All mares survived over follow-up periods of 6 months to 5 years and were used for riding (6 mares) and embryo transfer (1 mare, after partial ovariohysterectomy). CONCLUSIONS AND POTENTIAL RELEVANCE: According to this study, the prognosis for mares after ovariohysterectomy appears to be good, despite the technical difficulties of the procedure. The prevalence of life threatening complications can be lower than reported. PMID- 14756369 TI - Collagenase-1 (MMP-1) activity in equine synovial fluid: influence of age, joint pathology, exercise and repeated arthrocentesis. AB - REASONS FOR PERFORMING STUDY: Matrix metalloproteinases (MMPs) are considered candidate biomarkers for both physiological and pathological tissue remodelling because of their key role in articular cartilage homeostasis. As disruption of the collagenous architecture is thought to be pivotal in chronic degenerative diseases such as osteoarthritis (OA), the collagenases form an interesting subset of the MMPs. The significance of any biomarker in synovial fluid (SF) can be assessed properly only when fluctuations in patterns induced by physiological processes such as development and growth, and by external influences and interventions such as exercise and repeated arthrocentesis, are known and taken into account. OBJECTIVES: To investigate the activity of MMP-1 in equine SF at different stages of development and in joints affected by OA, and the influence of exercise and repeated arthrocentesis thereon. METHODS: MMP-1 activity was determined in SF of normal joints of fetal, juvenile and mature horses, and in SF of horses suffering from OA, using an internally quenched fluorogenic peptide substrate. MMP-1 activity was also measured in SF from horses subjected to an exercise regimen and those subjected to repeated arthrocentesis. RESULTS: An age related decline in the SF levels of active MMP-1 was observed. MMP-1 activity was 15-fold higher in fetal than in juvenile animals, which showed significantly higher MMP-1 activity levels than mature horses. In SF of OA joints, MMP-1 activity was increased. Exercise did not affect MMP-1 activity in SF, but repeated arthrocentesis (within 60 h) increased MMP-1 activity significantly. CONCLUSIONS: The high MMP-1 activity in SF of young individuals parallels the high metabolic activity occurring during rapid growth and differentiation at early age. The elevated MMP-1 activity in SF of OA joints probably reflects pathological matrix degradation, confirming the potential of MMP-1 to serve as a biochemical marker for early joint disease. Moderate exercise is not likely to influence the outcome of MMP-1 activity measurements in equine SF, but arthrocentesis should be taken into account as a possible confounding factor. POTENTIAL RELEVANCE: Given the crucial role of the collagen matrix for tissue integrity, MMP-1 activity may be a useful tool in diagnostic, therapeutic or prognostic studies in horses suspected of OA. However, care should be taken to exclude fluctuations in MMP-1 activity induced by physiological processes such as development and growth, and by interventions such as repeated arthrocentesis. PMID- 14756370 TI - Tissue-specific dysregulation of cortisol metabolism in equine laminitis. AB - REASONS FOR PERFORMING STUDY: The role of glucocorticoids (GCs) in the pathogenesis of laminitis is incompletely understood. Local tissue activity of GC is regulated by the steroid converting enzyme, 11beta-hydroxysteroid dehydrogenase-1 (11beta-HSD-1). Changes in integumentary (skin and hoof lamellar) 11beta-HSD activity occurring during laminitis could affect the extent to which GCs are involved in its development. HYPOTHESIS: That changes in integumentary 11beta-HSD-1 activity associated with the laminitic condition would lead to elevated local tissue levels of GCs, which could subsequently contribute, through paracrine and autocrine mechanisms, to the further development of laminitis; and that similar changes in 11beta-HSD-1 activity would be evident in both skin and hoof lamellar tissue. METHODS: Activity of 11beta-HSD-1 was determined in skin and hoof lamellar tissue specimens obtained from normal and laminitic horses using a radiometric assay. Skin samples were obtained from 10 normal horses and from 10 horses before and after induction of acute laminitis following administration of starch via nasogastric tube. Hoof lamellar samples were obtained from 10 normal horses, 10 horses following induction of acute laminitis and 4 chronically-foundered horses. Bidirectional 11beta-HSD-1 activity was measured in both skin and lamellar tissues. RESULTS: 11-ketoreductase activity exceeded 11beta-dehydrogenase activity in both skin and lamellar tissues. Cutaneous activity was higher than lamellar 11beta-HSD-1 activity in all groups. Both ketoreductase and dehydrogenase activity increased in skin and lamellae following experimental induction of acute laminitis, but the increase in ketoreductase activity was substantially greater than that for dehydrogenase in the lamellae. Induction of acute laminitis was attended by increases of 227 and 220% in cutaneous dehydrogenase and ketoreductase activity, respectively, and 173 and 398% in lamellar dehydrogenase and ketoreductase activity, respectively (P<0.05). CONCLUSIONS: The 11-ketoreductase moiety of 11beta-HSD-1 plays a role in equine skin and hoof lamellae regarding the regulation of local glucocorticoid activity. Increased 11-ketoreductase activity will lead to increased local tissue GC activity by virtue of conversion of cortisone to cortisol. POTENTIAL RELEVANCE: The laminitic condition is attended by integumentary biochemical changes that enhance the local concentration of cortisol, especially in the hoof lamellar interface. Through multiple and diverse actions, increased local GC activity contributes to the pathogenesis and morbidity associated with laminitis. Pharmacological manipulation of 11beta-HSD-1 deserves further investigation regarding the prevention and treatment of laminitis. PMID- 14756371 TI - Idiopathic muscular hypertrophy of the oesophagus in the horse: a retrospective study of 31 cases. AB - REASONS FOR PERFORMING STUDY: To present the first report of a case series concerning equine idiopathic muscular hypertrophy of the oesophagus (IMHO). OBJECTIVES: To investigate the clinical and pathological features of the disorder. METHODS: The medical records of 31 horses suffering from the disorder were reviewed retrospectively. In all these animals the diagnosis was confirmed at post mortem examination. RESULTS: The median age of the affected horses was 12.5 +/- 5.6 years (range 1-26) without sex or breed predilection. Only 2 out of 31 horses showed clinical signs associated with oesophageal dysfunction, indicating that the muscular hypertrophy was rather a coincidental post mortem finding. Histology revealed thickening of the distal portion of the oesophagus mainly involving the circular layer of the tunica muscularis without fibrosis or inflammation. In 8 cases, the disorder was seen in concurrence with idiopathic hypertrophy of the tunica muscularis of various other parts of the gastrointestinal tract. CONCLUSIONS: In the majority of patients, IMHO was a coincidental finding at post mortem examination usually confined to the smooth, circular muscle layer of the tunica muscularis externa. POTENTIAL RELEVANCE: Further research is necessary to study the precise effect of IMHO on oesophageal function. PMID- 14756372 TI - Transcranial magnetic stimulation: normal values of magnetic motor evoked potentials in 84 normal horses and influence of height, weight, age and sex. AB - REASONS FOR PERFORMING STUDY: Cervical spinal cord dysfunction is a common problem in equine medicine and the currently available tests give no objective information about the functionality of the nervous tracts. Therefore, transcranial magnetic stimulation (TMS) was performed in 84 healthy horses of different height in order to have an objective measure for the integrity of the descending motor tracts in normal horses. OBJECTIVES: To obtain reference values for onset latency and peak-to-peak amplitude of magnetic motor evoked potentials (MMEPs) and to evaluate the possible effect of height, age and gender on the neurophysiological measures. METHODS: All horses were sedated and stimulated transcranially by using a magnetic coil placed on the forehead. The stimulator triggered the sweep of an electromyogram machine that recorded MMEPs bilaterally from needle electrodes in the extensor carpi radialis and cranial tibial muscles. In that way, it was possible to measure latency between stimulus and onset of response. RESULTS: A significant difference was found between recordings made in the fore- and hindlimbs; MMEPs recorded in the front legs had a shorter onset latency and higher peak-to-peak amplitude. Mean +/- s.d. normal values for onset latency of 19.32 +/- 2.50 and 30.54 +/- 5.28 msecs and peak-to-peak amplitude values of 9.54 +/- 3.73 and 6.62 +/- 3.62 mV were obtained for extensor carpi radialis and cranial tibial muscles, respectively. The left-to-right difference in onset latency and peak-to-peak amplitude was not significant. In the same horse, differences up to 0.82 and 1.53 msecs for the extensor carpi radialis and cranial tibial muscles, respectively, lie within the 95% confidence limit and are considered normal. In contrast to onset latency, peak-to-peak amplitude showed a very large intra- and interindividual variability, even in the same muscle. To reduce the variability and predict normal values of new individual cases, influence of height, weight, age and sex on the MMEPs were determined. No significant effects of sex were observed on onset latency and peak-to-peak amplitude. The age of the horse had only a small but significant effect on peak to-peak amplitude, with larger responses in older horses. Height at the withers and weight of the horse, parameters that strongly correlate with the size of the horse, had an important significant influence on onset latency but not on peak-to peak amplitude. The age of the horse and height at the withers were used to predict peak-to-peak amplitude and onset latency, respectively, in normal horses. CONCLUSIONS AND POTENTIAL RELEVANCE: TMS is an excellent addition to the few tools we have for noninvasive imaging of the equine nervous system. Magnetically evoked potentials are highly reproducible and recent advances suggest that the applications of TMS in horses will continue to grow rapidly. PMID- 14756373 TI - A dose titration of triamcinolone acetonide on insulin-like growth factor-1 and interleukin-1-conditioned equine cartilage explants. AB - REASONS FOR PERFORMING STUDY: Previous in vitro pilot studies have defined a potentially beneficial effect of insulin-like growth factor-1 (IGF-1) and triamcinolone acetonide (TA) on interleukin-1 (IL-1)-conditioned equine cartilage. Furthermore, an optimal dose for IGF-1 treatment alone has been documented previously using the same test system as in the current project. OBJECTIVES: To perform a dose titration of TA on IL-1-conditioned equine articular cartilage explants in the presence of an optimised IGF-1 dose, in order to optimise a triamcinolone concentration for use in combination with IGF-1 for future investigations. METHODS: Cartilage explants were harvested from the distal femur of a normal horse. The effect of a clinically relevant TA dose range was evaluated in the presence of IL-1 and IGF-1 through measurement of proteoglycan (PG) matrix metabolism (synthesis and degradation). RESULTS: TA and IGF-1 in combination inhibited the IL-1-induced release of PG matrix components (glycosaminoglycan or GAG) from the articular cartilage, as well as producing a significant increase in GAG synthesis. CONCLUSIONS: This experiment provided proof of principle that a combination treatment appears to be able to combat the IL-1-induced matrix depletion, while enhancing anabolic metabolism within the articular cartilage. POTENTIAL RELEVANCE: The use of IGF-1 in conjunction with TA in vivo has the potential to provide beneficial anabolic effects not seen with TA alone. PMID- 14756374 TI - Is isoflurane safer than halothane in equine anaesthesia? Results from a prospective multicentre randomised controlled trial. AB - REASONS FOR PERFORMING STUDY: Approximately 1 in 100 horses suffer unexpectedly from anaesthetic-related death. Identification and use of the safest anaesthetic drugs should support this aim. Experimental evidence has suggested that isoflurane should be a safer maintenance agent in equine anaesthesia than halothane. HYPOTHESIS: The death rate would be reduced in horses being maintained with isoflurane compared to halothane. METHODS: A multicentre randomised controlled trial was undertaken to compare the effects of isoflurane and halothane for maintenance of equine anaesthesia for all types of operation. Data were analysed from 8242 horses in which anaesthesia was maintained with either halothane or isoflurane using mixed effects logistic regression models. RESULTS: No overall benefit of either drug was detected. However, although not part of the primary hypothesis, data showed that the overall death rate was significantly reduced in horses age 2-5 years with isoflurane and that death from cardiac arrest was also reduced with isoflurane, particularly in high risk cases. CONCLUSIONS AND POTENTIAL RELEVANCE: Halothane remains an acceptable anaesthetic for maintenance of anaesthesia in horses, but isoflurane may be safer in the young horse and in high risk cases. PMID- 14756375 TI - Equine recurrent uveitis is strongly associated with the MHC class I haplotype ELA-A9. PMID- 14756376 TI - Preliminary study of use of a polypectomy snare to obtain large samples of the equine gastric antrum by endoscopy. PMID- 14756377 TI - Equine herpesvirus-1 abortion: atypical cases with lesions largely or wholly restricted to the placenta. PMID- 14756378 TI - Equine degenerative myeloencephalopathy in five Quarter Horses: clinical and neuropathological findings. PMID- 14756379 TI - Acquired cervical scoliosis in six horses associated with dorsal grey column chronic myelitis. PMID- 14756380 TI - Fibrous dysplasia in the accessory carpal bone of a horse. PMID- 14756381 TI - Review of NMDA antagonist-induced neurotoxicity and implications for clinical development. AB - NMDA receptor antagonists have been investigated for many years as therapeutic agents for the treatment of neurological disorders such as stroke, epilepsy, pain and Parkinson's disease. It has been discovered, however, that many of these compounds cause adverse behavioral (psychotomimetic) effects and can produce neurotoxicity characterized by neuronal vacuolization, induction of heat-shock protein, neuronal/axonal degeneration and regional brain cell death in several animal species. It is unknown whether NMDA antagonists induce neurotoxicity in humans. The mechanism of NMDA antagonist-induced neurotoxicity is not completely known, but some evidence suggests disinhibition of GABAergic inputs to the affected neurons. Several classes of compounds have been shown to prevent NMDA antagonist-induced neurotoxicity. The extent of neurotoxicity produced by NMDA antagonists is affected by many factors, including type of antagonist, dose, length of exposure, age, sex and species. While there are no published regulatory guidelines regarding how NMDA antagonist compounds should be evaluated, sponsors and investigators of these compounds should make every effort to assess the potential for neurotoxicity. NMDA receptor antagonists, as well as other CNS active compounds need to be analyzed for neurotoxicity through careful experimental design, adequate tissue sampling and through the use of a sensitive method of detection. PMID- 14756382 TI - Programming of a flexible computer simulation to visualize pharmacokinetic pharmacodynamic models. AB - Teaching pharmacokinetic-pharmacodynamic (PK/PD) models can be made more effective using computer simulations. We propose the programming of educational PK or PK/PD computer simulations as an alternative to the use of pre-built simulation software. This approach has the advantage of adaptability to non standard or complicated PK or PK/PD models. Simplicity of the programming procedure was achieved by selecting the LabVIEW programming environment. An intuitive user interface to visualize the time courses of drug concentrations or effects can be obtained with pre-built elements. The environment uses a wiring analogy that resembles electrical circuit diagrams rather than abstract programming code. The goal of high interactivity of the simulation was attained by allowing the program to run in continuously repeating loops. This makes the program behave flexibly to the user input. The programming is described with the aid of a 2-compartment PK simulation. Examples of more sophisticated simulation programs are also given where the PK/PD simulation shows drug input, concentrations in plasma, and at effect site and the effects themselves as a function of time. A multi-compartmental model of morphine, including metabolite kinetics and effects is also included. The programs are available for download from the World Wide Web at http:// www. klinik.uni-frankfurt.de/zpharm/klin/ PKPDsimulation/content.html. For pharmacokineticists who only program occasionally, there is the possibility of building the computer simulation, together with the flexible interactive simulation algorithm for clinical pharmacological teaching in the field of PK/PD models. PMID- 14756383 TI - Pharmacokinetics of moxifloxacin are not influenced by a 7-day pretreatment with 200 mg oral itraconazole given once a day in healthy subjects. AB - AIM: The primary objective of this interaction study was to confirm preclinical data suggesting that moxifloxacin is not metabolized by CYP 450 isozymes. Itraconazole, a strong CYP 3A4 inhibitor, was used as comedication. METHODS: Twelve healthy male subjects were enrolled in this randomized study using 400 mg of oral moxifloxacin (MXF) administered alone and on the 7th day of a 9-day treatment regimen with itraconazole (ITR) 200 mg p.o., o.d. In addition to the assessment of safety and tolerability, non-compartmental pharmacokinetics of moxifloxacin, itraconazole and their respective metabolites were analyzed using plasma concentrations obtained using HPLC. RESULTS: All treatment regimens were safe and well-tolerated. No interaction with itraconazole was observed for moxifloxacin (relative bioavailability: 111.6% (90% CI 106.5 to 117.0%), C(max) ratio: 103.7% (84.8-126.9%) and its sulfometabolite (Ml) (AUC ratio: 107.7% (95.6, 121.4%), C(max) ratio: 105.8% (89.9-124.5%)). There was a 30% decrease in AUC with M2 moxifloxacin metabolite (glucuronide) accompanied by an approximately 54% increase in renal excretion, which may be due to changes in phase 2 metabolism and/or transport mechanisms altered by itraconazole. Exposure (AUC) to itraconazole and its hydroxymetabolite were marginally altered by moxifloxacin (AUC +5% for itraconazole and -5% for hydroxy-itraconazole (OH-ITR)) indicating the absence of a clinically relevant influence of moxifloxacin on itraconazole. Mean peak concentrations in plasma (C(max)) were reduced for ITR and OH-ITR by approximately 14% and 18%, respectively, when administered concomitantly with moxifloxacin. This was attributed to the sensitivity of itraconazole absorption to changes in gastric physiology (pH, gastric transit, administration after fasting) and was deemed as clinically irrelevant. CONCLUSION: Results of this study indicate that moxifloxacin is not a substrate for CYP 450 3A4 isozymes confirming previous preclinical in vitro data. Moxifloxacin can therefore be safely coadministered with CYP 3A4 inhibitors without the need for dose adjustment. No clinically relevant changes in the pharmacokinetics of itraconazole were observed during the study. PMID- 14756384 TI - Effects of recombinant growth hormone therapy on thyroid hormone concentrations. AB - BACKGROUND AND OBJECTIVE: There are numerous, often contradictory reports on the effects of growth hormone (GH) therapy on thyroid function. The aim of this study was to assess the effect of such therapy on serum concentrations of thyroid hormones in GH-deficient children euthyroid prior to the treatment, and to determine the necessity of thyroid hormone administration in these patients. MATERIAL AND METHODS: The study included 32 GH-deficient patients in the first stage of sexual development, in whom disorders of thyroid function could be excluded. The inclusion criteria were based on clinical examination and levels of thyroxine (T4), triiodothyronine (T3), free thyroxine (fT4), free triiodothyronine (fT3), reverse triiodothyronine (rT3), thyrotropin (TSH) before and after stimulation with thyrotropin-releasing hormone (TRH). Recombinant growth hormone (rGH) (Genotropin 16U, Pharmacia) was administered at a dose of 0.7 U/kg/week. Fasting blood samples were drawn before treatment and after 3, 6, 9 and 12 months of therapy. Thyroid hormones were measured using RIA and IRMA methods. RESULTS: There were no physical signs of hypothyroidism in the patients examined during 12 months of rGH administration, and the satisfactory growth rate was achieved. T4 levels decreased in the first 3 months but remained within the normal range, and then returned to the values prior to the treatment. A similar trend was observed for fF4, with 28.5% of patients exhibiting fF4 levels below the normal in the 3rd month. An increase during the first 3 months of therapy was observed in the cases of T3 (statistically non-significant) and fT3, and these values then fell to levels within the normal range of patients' age. During treatment, TSH levels decreased but remained within the normal range. CONCLUSIONS: A transient decrease in T4 concentrations in the 3rd month with unchanged T3 and an increase in fT3 concentrations probably result from the effect of rGH on the peripheral metabolism of thyroid hormones. The results obtained do not support the use of thyroid hormone therapy with levothyroxine during the first year of rGH therapy in patients who are initially euthyroid. PMID- 14756385 TI - Presurgical antimicrobial prophylaxis: effect on ocular flora in healthy patients. AB - OBJECTIVE: Evaluation of presurgical antimicrobial prophylaxis for reduction of ocular flora. SETTING: Ophthalmology Section, Department of Medicine and Surgery, University of Catania, Italy. METHODS: Three days before photorefractive keratectomy (PRK), conjunctiva of 70 healthy patients (100 eyes) were swabbed. After 3 days of instillation of ofloxacin 0.3% (3 times daily), conjunctival swabs were taken again. Bacteria were isolated and identified from each swab. RESULTS: A total of 191 independent isolates were obtained prior to antimicrobial treatment. Bacteria were isolated from all 100 eyes sampled. Gram-positive species predominated, with Staphylococcus epidermidis and Staphylococcus aureus cultured from 69% and 25% of eyes sampled, respectively. Species of the anaerobic genera Peptococcus and Peptostreptococcus were found in 22% and 14% of eyes, respectively. After prophylaxis with ofloxacin, bacteria could be cultured from only 7% of eyes (10 independent isolates). All isolates were sensitive to ofloxacin except 5 strains of S. epidermidis, which displayed intermediate sensitivity. No infections occurred after PRK. CONCLUSION: Prophylaxis by instillation of ofloxacin 0.3% 3 times daily for 3 days substantially reduced the ocular flora of 100 healthy eyes prior to refractive surgery. PMID- 14756386 TI - Bioequivalence study of two fluconazole capsule formulations in healthy volunteers. AB - OBJECTIVE: To compare the bioavailability of a fluconazole 150 mg capsule formulation from Laboratorio Teuto Brasileiro Ltd., Brazil (test formulation), and Zoltec 150 mg capsule from Laboratorios Pfizer Ltd., Brazil (reference formulation), in 24 volunteers of both sexes. MATERIAL AND METHODS: The study was conducted open with randomized 2-period crossover design and a 2-week washout period. Plasma samples were obtained over a 168-hour interval. Fluconazole concentrations were analyzed by combined reversed-phase liquid chromatography and tandem mass spectrometry (LC/MS/MS) with positive ion electrospray ionization using selected ion monitoring method. From the fluconazole plasma concentration vs. time curves the following pharmacokinetic parameters were obtained: AUC(last), AUC(0-inf) and C(max). RESULTS: Geometric mean of fluconazole/Zoltec 150 mg individual percent ratio was 102.6% for AUC(last), 102.2% for AUC(0-inf) and 109.4% for C(max). The 90% confidence intervals were 97.3-108.2%, 97.0 107.8%, and 103.1-116.0%, respectively. CONCLUSION: Since the 90% CI for both Cmax, AUC(last) and AUC(0-inf) were within the 80-125% interval proposed by the Food and Drug Administration, it was concluded that fluconazole 150 mg capsule was bioequivalent to Zoltec 150 mg, according to both the rate and extent of absorption. PMID- 14756387 TI - Phentolamine bioequivalence study. AB - OBJECTIVE: To assess the bioequivalence of 2 tablet formulations of phentolamine (Regitine phentolamine 40 mg tablet formulation by Novartis, Brazil, as test formulation, and Vasomax, phentolamine 40 mg tablet formulation by Schering Plough S.A., Brazil, as reference formulation). METHODS: A single 40 mg oral dose of each formulation was administered to 36 male healthy volunteers. The study was conducted after screening, using an open, randomized, 2-period crossover design, a 7-day interval between doses, and wash-out period of at least 4 weeks. Plasma samples for determination of phentolamine were obtained predose and at intervals over 720 min postdose. Plasma concentrations were quantified by reversed-phase liquid chromatography coupled to tandem mass spectrometry (LC-MS-MS) with positive ion electrospray ionization using multiple reactions monitoring (MRM) method. Precision of the method was evaluated using calibration curves and plasma quality control samples. The subjects were monitored throughout the study. Systolic and diastolic blood pressure and pulse rate measurement were taken predose and at intervals up to 720 min. Tolerance of both products was good. No serious adverse reactions were reported. The pharmacokinetic parameters calculated for both compounds included: AUC(0-720 min), AUC(0-infinity), C(max), Ca and k(e). RESULTS: The maximum concentrations reached (C(max)) were compared. Regitine 40 mg formulation C(max) geometric mean ratio was 108.29% (90% CI = 98.58-118.96) of Vasomax 40 mg formulation. The areas under the curve (AUC(0-720 min)) were compared. Regitine 40 formulation (AUC(0-720 min)) geometric mean ratio was 102.33% (90% CI = 97.21-107.72) of Vasomax 40 mg formulation. CONCLUSION: Since the 90% CI for both C(max) and AUC ratio where inside the 80 to 125% interval proposed by the Food and Drug Administration, it is concluded that Regitine 40 mg tablet is bioequivalent to Vasomax for the rate and extent of absorption. PMID- 14756388 TI - Bioequivalence study comparing a new paracetamol solution for injection and propacetamol after single intravenous infusion in healthy subjects. AB - OBJECTIVES: A new, ready-to-use solution for injection of paracetamol (Perfalgan 10 mg/ml) without previous reconstitution has been developed. The aim of the study was to determine the serum concentration profiles of paracetamol after 15 min infusion of Perfalgan 0.5 g and 1 g doses and to demonstrate the bioequivalence between Perfalgan 1 g dose and a marketed reference formulation for injection, propacetamol 2 g (Pro-Dafalgan 2 g) equivalent to 1 g of paracetamol. The secondary objective was to evaluate local tolerance, and clinical and biological safety. METHODS: The study was performed in 24 healthy, male volunteers, according to an open-label, randomized, single-dose, 3-period crossover design, with a 1-week washout period between the doses. Blood samples were taken prior to each administration and at 18 time points within the 24-hour period following the beginning of each infusion. Serum concentrations of paracetamol were determined by validated high-performance liquid chromatography with UV detection. From serum concentration-time data, a non-compartmental pharmacokinetic analysis was performed to calculate Cmax, tmax, AUC(inf), t(1/2), MRT, Cl(T) and Vd. Log-transformed AUC(inf) and Cmax were tested for bioequivalence. The local pain intensity at infusion site was assessed using a 4 point categorical scale from 0 (none) to 3 (severe). The clinical and biological safety was evaluated by physical examination with measurements of vital signs and ECG and laboratory tests including hematology and biochemistry. RESULTS: After infusion of 0.5 g and I g of the new paracetamol solution, C(max) and AUC(inf) increased proportionally with dosage. After dose correction to 1 g of paracetamol, the mean (+/- SD) Cmax ratio was 0.98 +/- 0.24 and 0.94 +/- 0.08 for AUC ratio. Identical t(max) was observed for the 2 paracetamol dosages and 90% confidence intervals for t(1/2), MRT, Cl(T) and V(d) were within the acceptable interval 0.8-1.25. The calculated 90% confidence intervals of the new solution (Perfalgan 1 g) to marketed solution (propacetamol 2 g) ratios were 1.11-1.31 (point estimate 1.20) for C(max) and 1.10-1.16 (point estimate 1.13) for AUC(inf). These values were within the acceptable bioequivalence intervals of 0.75 to 1.33 for Cmax and 0.80-1.25 for AUC(inf). Application site disorders were the most frequently observed adverse events but local pain at infusion site was less reported by subjects after Perfalgan (2%) compared to propacetamol (20%). The clinical and biological safety was good and equivalent for the 3 treatments. CONCLUSION: After administration of paracetamol solution for injection 0.5 g and 1 g, the pharmacokinetics of paracetamol is linear. All results indicate that 1 g of paracetamol administered as Perfalgan 10 mg/ml is bioequivalent to propacetamol 2 g with a better local safety. PMID- 14756389 TI - Reproducibility of nifedipine absorption from GITS tablets: comparison of single dose pharmacokinetics using 10, 20, 40 and 60 mg nifedipine. AB - OBJECTIVE: To examine the reproducibility of nifedipine absorption from gastrointestinal therapeutic system (GITS) tablets by comparing the single-dose pharmacokinetic profiles of 4 different dosages administered orally. METHODS: Twelve healthy male volunteers, aged between 22 and 29 years were enrolled in the open, 4-way, dose escalation study with single oral doses of 10, 20, 40 and 60 mg (two 30 mg) nifedipine GITS tablets. Each administration was separated by a 1 week washout period. Coefficients of variation (CV) of dose-corrected area under the concentration-time curve (AUC) and peak plasma drug concentrations (Cmax) were calculated from the pharmacokinetic profiles. RESULTS: Mean AUC and mean Cmax were dose-proportional from 10 to 60 mg. Although the CV of 4 mean dose corrected AUC and Cmax were 5.5% and 17.5%, respectively, CV of dose-corrected AUC and Cmax in each subject varied from 5.1 to 37.4% (mean 11.0%) and from 14.1% to 46.4% (mean 25.8%), respectively. CONCLUSIONS: Whereas mean plasma nifedipine concentration remained markedly stable over a 16- to 24-hour interval and mean dose-corrected AUC showed good reproducibility with nifedipine GITS, the CV of dose-corrected AUC of the nifedipine GITS tablets in each subject showed large variability. PMID- 14756390 TI - Adaptive learning in arthropods: spider mites learn to distinguish food quality. AB - Many herbivorous arthropods have been shown to possess learning capabilities, yet fitness effects of learning are largely unknown. In this paper, we test whether two-spotted spider mites (Tetranychus urticae) learn to distinguish food quality in choice tests, and whether this results in fitness benefits. Food consisted of cucumber plants with one of three degrees of feeding damage: undamaged (no mites), mildly damaged (infested by a mite strain adapted to tomato) and heavily damaged (infested by a mite strain adapted to cucumber). Mites were subjected to one choice test in a greenhouse and three sequential choice tests on leaf disks. Thereafter, individual mite performance was measured as oviposition rate over four days. In the course of the three small-scale choice tests, preference shifted towards less damaged food. The performance tests showed that learning was adaptive: mites learned to prefer the food type that yielded the higher oviposition rate. Interestingly, innate preferences in the greenhouse tests were close to those shown after learning in the small-scale tests. Given that both strains of mites had not experienced cucumber for several years, we hypothesize that the preference in the greenhouse was due to avoidance of mite odours rather than odours of damaged plants. Through its effect on foraging behaviour, adaptive learning may promote the evolution of host plant specialization in herbivorous arthropods. PMID- 14756391 TI - The effects of clofentezine on life-table parameters in two-spotted spider mite Tetranychus urticae. AB - Sublethal effects of the growth inhibitor, clofentezine, on life-table parameters of Tetranychus urticae Koch females treated at different developmental stages with a concentration causing >90% mortality were investigated. Females which survived treatment as 'early' (0-24 h old) eggs produced 12% more offspring than the untreated females during the first five days of oviposition. This resulted in a significant rise in the intrinsic rate of increase (rj): 0.324, compared to 0.299 in the untreated females. This effect may be interpreted as hormoligosis. Clofentezine treatment at any other developmental stage of T. urticae significantly decreased both longevity and fertility of female survivors. Females which survived treatment either as 'late' (72-96 h old) eggs or larvae had 2.6 times lower net reproductive rate (R0) than the untreated females, and the rj values were significantly lower: 0.242 and 0.215, respectively (0.285 in the untreated females). Females which survived treatment either as protonymphs or deutonymphs had 3.9 times and 6 times lower R0, respectively. Corresponding rj values were 0.178 and 0.146, respectively (0.247 in the untreated females). The clofentezine treatment at all stages influenced the age distribution of survivors. The sublethal effects of clofentezine and their impact on T. urticae management are discussed. PMID- 14756392 TI - Effects of prey mite species on life history of the phytoseiid predators Typhlodromalus manihoti and Typhlodromalus aripo. AB - The effects of prey mite suitability on several demographic characteristics of phytoseiid predators and the relationship of these effects to the potential of phytoseiid predators to control herbivorous mite populations are well documented. Evidence has also accumulated in the last 20 years demonstrating that phytoseiid predators utilize herbivorous prey mite-induced plant volatiles as olfactory cues in locating their herbivorous mite prey. but less well established is the predictability of reproductive success from the ability of the predators to utilize olfactory cues to locate their prey, and how these processes are related to the success of the predators as biological control agents of the herbivorous mite. In this study, we determined in laboratory no choice experiments, the development, survivorship and fecundity of the two neotropical phytoseiid predators Typhlodromalus manihoti Moraes and T. aripo DeLeon when feeding on three herbivorous mites, including the key prey species Mononychellus tanajoa (Bondar), and the two alternative prey species Oligonychus gossypii (Zacher) and Tetranychus urticae (Koch). Intrinsic rate of increase (rm) of T. aripo was 2.1 fold higher on M. tanajoa as prey compared with T. urticae as prey, while it was almost nil on O. gossypii. For T. manihoti, rm was 2.3 fold higher on M. tanajoa as prey compared with O. gossypii as prey, while reproduction was nil on T. urticae. An independent experiment on odor-related prey preference of the two predator species (Gnanvossou et al. 2002) showed that T. manihoti and T. aripo preferred odors from M. tanajoa-infested leaves to odors from O. gossypii infested leaves. Moreover, both predator species preferred odors from M. tanajoa infested leaves over those from T. urticae-infested leaves. As reported here, life history of the two predatory mites matches odor-related prey preference if the key prey species is compared to the two inferior prey species. The implications of our findings for the persistence of T. manihoti and T. aripo and biological control of M. tanajoa in the cassava agroecosystem in Africa are discussed. PMID- 14756393 TI - Seasonal prevalence and susceptibility to agrochemicals of Tyrophagus similis (Acari: Acaridae) in spinach buds and agricultural soil under greenhouse conditions. AB - Seasonal prevalence of Tyrophagus similis was investigated from 1997 to 1998 in two spinach greenhouses in central Japan. Susceptibility of T. similis to agrochemicals was also tested in the laboratory. Tyrophagus similis density in the soil was low during the high temperature period from May to August. The density rapidly increased in late autumn and remained at a high level during the cool season from December to February. The number further increased in April and then rapidly decreased in May. The high temperatures in the greenhouses from spring to early autumn are considered the main causes of population decrease. Mites on spinach buds increased the number after those on and in the soil increased. Mites attacked spinach buds mostly in late autumn and early spring. Dichlorvos did not reduce the number of mites in either greenhouse even though it was highly toxic under laboratory condition. This discrepancy suggests that the mites in both the soil and spinach buds had little direct contact with the chemicals. These data suggest that once crop damage by mites is detected, it is usually too late to use chemicals, and that mites that live in the buds are protected from agrochemicals. PMID- 14756394 TI - Seasonal and diel activity of Ixodes ricinus (Acari: Ixodidae) subpopulations in Denmark. Aspects of size, physiological age, and malate dehydrogenase genotype in a forest site without any undergrowth. AB - The underlying population dynamics and the behavioural patterns of the vectors are key issues in understanding the transmission of vector borne pathogens. For the tick Ixodes ricinus both seasonal and diel activity have been described as bimodal patterns, which in seasonal aspect has been interpreted as representing two cohorts. However, recent studies have shown that this interpretation may be incorrect. The aim of this study was to obtain more detailed information on nymph host seeking by studying subpopulations of ticks during the day and season. The study was designed to allow for comparisons of the diel variation and seasonal variation in their dependency in a number of tick characteristics. The study took place in a forest with planted beech trees without any undergrowth. Ticks were collected by flagging the dead leaves on the forest floor. For each nymph, a number of visual observations were made. The size and physiological age was observed and the nymphs were genotyped in the malate dehydrogenase locus (MDH, E.C. 1.1.1.37). Briefly the main results can be given as: (i) There were significant differences in the composition of size classes during the season, but only limited trends in time. (ii) The proportion of the small nymphs was highly variable, with a variation from 3% to 24% in October and September, respectively. (iii) The diel variation in MDH genotypes was significant in May and August. (iv) Nymph size classes and physiological age appeared to interact. The non-random interaction was caused by a relatively even distribution of small nymphs in all four age classes, while large nymphs tended to fall into age class 2 and 3. The length by age interaction for the individual months was noted to be significant in May, July, August and September, but not in June. Similarly the interaction was significant in the morning and afternoon, but not at midday. The overall results describe the seasonal and diel activity patterns as changing systematically for several characteristics under the influence of weather condition and population dynamics. IN CONCLUSION: The observations are best interpreted as being produced of a single cohort of ticks, but the revealed complexity of the host seeking activity suggest that measures of activity x abundance should be interpreted very cautiously in relation to population dynamics. PMID- 14756395 TI - Life cycle and host specificity of Amblyomma triste (Acari: Ixodidae) under laboratory conditions. AB - We report biological data of two generations of Amblyomma triste in laboratory and compared the suitability of different host species. Infestations by larval and nymphal stages were performed on guinea pigs (Cavia porcellus), chickens (Gallus gallus), rats (Rattus norvegicus), rabbits (Oryctolagus cuniculus), wild mice (Calomys callosus), dogs (Canis familiaris) and capybaras (Hydrochaeris hydrochaeris). Infestations by adult ticks were performed on dogs, capybaras and rabbits. Tick developmental periods were observed in an incubator at 27 degrees C and RH 90%. Guinea pigs were the most suitable hosts for larvae and nymphs, followed by chickens. The remaining host species were less suitable for immature ticks as fewer engorged ticks were recovered from them. Mean larval feeding periods varied from 3.8 to 4.7 d between different host species. Mean larval premolt periods ranged from 8.9 to 10.4 d. Nymphal mean feeding periods varied from 4.2 to 6.2 d for ticks fed on different host species. Premolt period of male nymphs (mean: 15.4 d) was significantly longer than that of female nymphs (14.7 d). Female nymphs were significantly heavier than male nymphs. The overall sex ratio of the adult ticks emerged from nymphs was 0.9:1 (M:F). Capybaras were the most suitable host for the tick adult stage as significantly more engorged females were recovered from them and these females were significantly heavier than those recovered from dogs or rabbits. The life cycle of A. triste in laboratory could be completed in an average period of 155 d. The potential role of guinea pigs, birds and capybaras, as hosts for A. triste in nature, is discussed. PMID- 14756396 TI - Propensity towards cannibalism among Hypoaspis aculeifer and H. miles, two soil dwelling predatory mite species. AB - In biological control programmes, the two predatory soil mites Hypoaspis aculeifer and H. miles are often applied against soil-borne pests like mushroom flies, springtails and mites. Although the mites show high consumption rates on varying prey types in Petri dish experiments and in greenhouses, their overall efficiency is sometimes limited. We hypothesized that intraspecific interactions, like cannibalism, could contribute to this decreased competence. Therefore, experiments were conducted to show the propensity of H. aculeifer and H. miles to cannibalise. Adult mites and nymphs were introduced as predators with conspecific eggs, larvae, nymphs, adult females or males as prey and the number of killed individuals was recorded. Additionally, the oviposition rate on conspecific prey was quantified and the correlation with the number of prey consumed was calculated to assess the influence of cannibalism on egg production. The results illustrate that cannibalism occurs infrequently in both Hypoaspis spp., the only exception being H. aculeifer nymphs, which cannibalised one conspecific egg per day. Moreover, cannibalism never occurred in the presence of alternative prey. Oviposition rate decreased during the experiment in both species but it was positively correlated with the cannibalism rate only for H. aculeifer. The benefit of cannibalism for populations of H. aculeifer and H. miles is discussed. PMID- 14756397 TI - Phytoseiid predators of whiteflies feed and reproduce on non-prey food sources. AB - Two phytoseiid species, Euseius scutalis (Athias-Henriot) and Typhlodromips swirskii (Athias-Henriot), are able to suppress whitefly populations on single plants and are candidate biological control agents for whiteflies such as Bemisia tabaci (Gennadius). These species can feed on pollen and insect-produced honeydew and these food sources are likely to be available in crops. If the utilization of these food types results in increased reproduction or survival, populations of predators can persist when whitefly prey is scarce or absent. We studied the impact of pollen and whitefly-produced honeydew on the life history of the two phytoseiids. Cattail pollen allowed for survival, development and reproduction of both predators. Whitefly-produced honeydew greatly increased survival of E. scutalis, allowed for development into adulthood and for a sustained low rate of oviposition. The survival of adult T. swirskii was high on cucumber leaf tissue, either with or without pollen or honeydew. Oviposition by adults and juvenile survival of T. swirskii was very low in presence of honeydew. Biological control of whiteflies may benefit from both pollen and honeydew because these non-prey food sources have a positive effect on the life history of the two predator species, especially E. scutalis. PMID- 14756399 TI - The effect of temperature on the functional response of Phytoseiulus persimilis (Acari: Phytoseiidae). AB - Environmental variables, such as temperature, are important in determining the efficiency of biological control in ornamental crops. This paper examines the effect of temperature on the functional response of adult female Phytoseiulus persimilis to eggs of the spider mite, Tetranychus urticae. The functional response was determined using a new functional response assay technique with plant stems as an arena, rather than leaf discs. The use of plant stems allows the influence that plant structure has on predation to be incorporated into the assay. Control assays were also used (without predators) to estimate natural losses of prey. The data were analysed using a binomial model, with the use of Abbot's formula to correct for the losses in the controls. A combined equation to describe the effect of temperature and prey density on the predation rate of Phytoseiulus persimilis was derived. The results showed that more prey are eaten as the temperature increases from 15 degrees C to 25 degrees C, but the number of prey eaten then declines at 30 degrees C, although not to the levels seen at 20 degrees C. The implication of these results for biological control in ornamental crops, where the temperature can often exceed 30 degrees C, is discussed. PMID- 14756398 TI - Phytoseiid predator of whitefly feeds on plant tissue. AB - Predatory mites of the family Phytoseiidae feed on herbivorous mites and insects but they also use a variety of non-prey food items, such as pollen and nectar. Plant tissue is another potential food source. We investigated whether plant feeding occurs in the two phytoseiids Euseius scutalis (Athias-Henriot) and Typhlodromips swirskii (Athias-Henriot), which are natural enemies of whiteflies. These predatory mites can suppress populations of Bemisia tabaci (Gennadius) on isolated plants and are candidates for biological control of this pest. Both species can be reared on a diet of pollen, but E. scutalis requires a leaf tissue substrate, suggesting that this species might feed on plant tissue. To test this hypothesis, we applied a systemic insecticide (aldicarb) to cucumber plants and assessed the survival of predatory mites on leaves from insecticide-treated plants and untreated plants, both in presence and absence of pollen. The survival of T. swirskii was not affected by the presence of systemic insecticide in the plant. However, the survival of E. scutalis on leaves from insecticide-treated plants was 10 times lower than on leaves from untreated plants. Since the two species showed similar sensitivity to the insecticide when applied in a slide-dip test, this suggests that E. scutalis ingested insecticide through feeding on the leaf tissue. Mortality on treated leaves was observed both in absence and presence of pollen, suggesting that plant feeding is indispensable for E. scutalis. The extent to which plants are used as food by E. scutalis requires further analysis. PMID- 14756400 TI - Side-effects of three pesticides on the predatory mite, Phytoseiulus persimilis (Acari: Phytoseiidae). AB - Side-effects of three commonly used pesticides in Iran were evaluated on an introduced strain of the predatory mite. Phytoseiulus persimilis Athias-Henriot, reared for about 10 years without exposure to any pesticides. Application of pesticides was carried out either to detached bean leaves using a Potter tower at 1 mg wet deposit per cm2 or by a hand sprayer on bean plants until run off. According to an EPPO decision making scheme, pirimiphos-methyl was found to be harmful (E=90.8%) and heptenophos harmless (E=-3.7%) to the predatory mite in the residual initial toxicity tests. For determination of the hazard class of malathion a field test was found to be necessary (E=59.8%). Categories of 1, 2, 3 were determined for heptenophos, malathion and primiphos-methyl, respectively, using IOBC classification. Despite being harmful, it is possible to use pirimiphos-methyl 10 days before release of P. persimilis. Investigation of the contribution of both lethal and sub-lethal effects to total impact indicated the dominance of lethality in the case of pirimiphos-methyl, while malathion acted by both mechanisms. Heptenophos did not have negative effects on fecundity of P. persimilis but rather caused a higher rate of fecundity in comparison with the control. The mortality found in the heptenophos test was not significantly different from the control. PMID- 14756401 TI - Ambulatory migration in mites (Acari: Tetranychidae, Phytoseiidae) to new leaves of moso bamboo shoots. AB - The migratory behaviour of two tetranychid pest species, Aponychus corpuzae and Schizotetranychus nanjingensis, and one phytoseiid, Typhlodromus bambusae, was studied in several monocultural bamboo forests in Fujian Province, China. The aim of the study was to assess how the ambulatory immigration of tetranychid and phytoseiid mites from the ground to new leaves is affected by a sticky barrier around the stem, by the age of bamboo shoots or by shoot density. The results show that while the sticky barrier is particularly effective at disrupting the ambulatory immigration from the ground to new leaves of S. nanjinigensis to 1 year-old shoots and of A. corpuzae to 3-year-old shoots, it has no significant effect on the immigration of the phytoseiid mite. PMID- 14756402 TI - Distribution patterns of coconut mite, Aceria guerreronis, and its predator Neoseiulus aff. paspalivorus in coconut palms. AB - Distribution patterns and numerical variability of the coconut mite Aceria guerreronis Keifer (Acari: Eriophyidae) and its predator Neoseiulus aff. paspalivorus DeLeon (Phytoseiidae) on the nuts of 3- to 7-month-old bunches of coconut palms were studied at two sites in Sri Lanka. At the two sites, coconut mites were present on 88 and 75% of the nuts but no more than three-quarters of those nuts showed damage symptoms. N. aff. paspalivorus was found more on mature nuts than on immature nuts. Spatial and temporal distribution of coconut mites and predatory mites differed significantly. The mean number of coconut mites per nut increased until 5-month-old bunches and declined thereafter. The densities of predatory mites followed a similar trend but peaked 1 month later. Variability in the numbers of mites among palms and bunches of the same age was great, but was relatively low on 6-month-old bunches. The results indicate that assessment of infestation levels by damage symptoms alone is not reliable. Sampling of coconut and/or predatory mite numbers could be improved by using several nuts of 6-month old bunches. The effect of predatory mites on coconut mites over time suggests that N. aff. paspalivorus could be a prospective biological control agent of A. guerreronis. PMID- 14756403 TI - Life history of hawthorn spider mite Amphitetranychus viennensis (Acarina: Tetranychidae) on various apple cultivars and at different temperatures. AB - Development duration and reproduction rate of hawthorn spider mite Amphitetranychus viennensis (Zacher) were carried out on five different apple cultivars (Amasya (local cultivar), Golden Delicious, Granny Smith, Starking Delicious and Starkrimson Delicious) at 25 degrees C, 65 +/- 10% RH and 16:8 L:D. In addition, the same parameters were determined on Golden Delicious leaves at three constant temperatures (20, 30 and 35 degrees C, 65 +/- 10% RH and 16:8 L:D) in the laboratory. A. viennensis showed a better performance on Golden Delicious than on the other apple cultivars. This was mainly due to a short development time (10.7 days), high daily egg production (5.2 eggs/female/day) and early reproduction peak. The highest intrinsic rate of natural increase (rm) was determined on the variety Golden Delicious (rm = 0.247/day), while the lowest one was observed on the variety Starking Delicious (rm = 0.215/day). The developmental periods of A. viennensis varied from 7.4 to 18.8 days at 35 and 20 degrees C for females, while it varied from 7.9 to 17.2 days at 30 and 20 degrees C for males. The development threshold of the eggs and pre-adult stages were 9.72 and 9.07 degrees C, total effective temperature was 72.99 and 185.18 degree-days, respectively. The mean generation time (To) of the population ranged from 16.13 days at 30 degrees C to 29.15 days at 20 degrees C. The net reproductive rate (R0) increased from 54.33 female/female at 20 degrees C to 78.34 female/female at 25 degrees C, and decreased to 75.71 female/female at 30 degrees C. The highest intrinsic rate of increase (rm) was reached at 30 C (rm = 0.268/day), the lowest one at 20 degrees C (rm = 0.136/day). PMID- 14756404 TI - Isolation, characterization, inheritance and linkage of microsatellite markers in Tetranychus kanzawai (Acari: Tetranychidae). AB - We isolated and characterized seven microsatellite markers in Tetranychus kanzawai (Acari: Tetranychidae). We also examined the conformity of the isolated markers to Mendelian laws and analyzed linkage among the microsatellite loci. All microsatellite markers fit expected 1:1 disomic segregation ratio and hence were inherited in a Mendelian manner. Significant pairwise linkage was detected in three pairs of microsatellite loci. These isolated microsatellite markers may become a powerful tool for the study of behavioral ecology, population genetics, and genome mapping of T. kanzawai. PMID- 14756405 TI - Microanatomical and biological aspects of bacterial associations in Tyrophagus putrescentiae (Acari: Acaridida). AB - The occurrence of bacterial colonies in the mesenchymal tissue of Tyrophagus putrescentiae Schrank was studied. One algal and two fungal species were offered as food. The experiment was analysed histologically and via transmission electron microscopy and plating of the mite homogenate. The dominant bacterium species, the bacterium population density in the mesenchyme, the location of bacterium groups between the internal organs and the ultrastructure were investigated. There were conspicuous differences between food types tested. On Penicillium, the highest population of bacteria in the mesenchymal tissue was correlated with the chitinase activity in mite homogenate. In this homogenate, Serratia marcescens was highly dominant among the other plated bacteria and it exhibited strong chitinolytic and trehalolytic activity. PMID- 14756406 TI - The effect of modified atmospheres on the survival of the eggs of four storage mite species. AB - The results of a laboratory investigation into the effects of modified atmospheres (MA) on the eggs of mite pests of grain and cheese are presented. Four species of astigmatid mite were tested; Acarus farris (Oudemans). A. siro L., Lepidoglyphus destructor (Schrank) and Tyrophagus longior (Gervais). All are found in many habitats including grain and cheese stores. Three low oxygen (O2) MA mixtures were used, based on carbon dioxide (CO2), nitrogen (N2) or simulated burner gas (0.5 or 2% O2, 10% CO2, balance N2) plus 60% CO2 in air (8% O2). The mites were exposed at 15 degrees C and 80% r.h., a combination of conditions that occurs at the surface of stored grain during the autumn which promotes mite population growth. The exposure periods required to prevent egg hatch for each species in every mixture are given. Tyrophagus longior was the most tolerant species, followed by A. siro and A. farris, with L. destructor the most susceptible. Burner gas was the most effective mixture overall with 0.5% O2 but with an increase in the O2 level to 2% for all the mixtures, CO2 became the more effective control agent. With 60% CO2 in air some loss of efficacy was observed against the three most tolerant species and even more so for L. destructor. Sublethal exposures to MAs for at least 4 days in L. destructor, 6 days in A. farris and A. siro and 8 days for T. longior caused a delay in egg hatch. PMID- 14756407 TI - First detection of chlorfenapyr (Secure) resistance in two-spotted spider mite (Acari: Tetranychidae) from nectarines in an Australian orchard. AB - Chlorfenapyr resistance (2.9- and 19.9-fold respectively at LC50 and LC99 level) was detected in Tetranychus urticae Koch causing control failure following a single application of product to nectarines. PMID- 14756408 TI - Inheritance of resistance to flumethrin in the Mexican Aldama strain of the cattle tick Boophilus microplus (Acari: Ixodidae). AB - The objective of this study was to evaluate the inheritance mode of resistance to flumethrin in the Mexican Aldama Boophilus microplus strain. Two Mexican strains were used, the Chiapas susceptible (SS), and the Aldama flumethrin-resistant from Tamaulipas. Six steers weighing ca. 250 kg were randomly assigned for each of six crosses: the susceptible (SS), resistant (RR), and the F1 (RS, SR) reciprocal crosses and F2 (RS x RS and SR x SR). The reciprocal crosses were made to evaluate maternal and sex linkage effects. Bioassays tested resistant and susceptible larvae along with their hybrid F1 and F2 progeny against a series of concentrations of flumethrin (0, 0.0075, 0.00150, 0.00300, 0.00600 and 0.01200 microg/g). To test the single-gene hypothesis of resistance, a nonparametric line cross test proposed by Collins was used. The bioassay data were subjected to probit analysis and the resistance factor and effective dominance obtained. Results of this study indicated that inheritance for flumethrin resistance in the Aldama strain was autosomal and controlled for more than one gene. The F1 and F2 larvae had similar lower resistant factor (RF 2.8-4.5) while the resistant Aldama strain was 21-fold higher (RF 81.8) than the mean of the F1 and F2. The extent of flumethrin resistance in the Aldama B. microplus strain depended upon the concentration of the pesticide used. Resistance was almost dominant at the lowest dose while almost completely recessive at the highest dose. Maternal effects were shown for egg-mass. These results shown here indicate more than one gene basis of flumethrin resistance in B. microplus ticks are present. Therefore it is necessary to locate and understand the major loci for elucidate the mechanism of resistance and improve the ability to track and delay the evolution of resistance. PMID- 14756409 TI - Ability of Ixodes persulcatus, Haemaphysalis concinna and Dermacentor silvarum ticks to acquire and transstadially transmit Borrelia garinii. AB - In China, the ability of Ixodes persulcatus, Haemaphysalis concinna and Dermacentor silvarum to transmit Borrelia spirochetes was determined under laboratory conditions. Results showed that all three tick species can acquire spirochetes by feeding on infected mice. However, the capability of the three species to maintain spirochetes was different. Only I. persulcatus is able to maintain spirochetes alive during molting, and subsequent tick stages transmitted the infection to naive mice. In H. concinna and D. silvarum ticks, spirochetes could not survive after the digestion period of blood and after the molting process was finished, spirochetes were no longer detected. Contrary to H. concinna and D. silvarum, I. persulcatus should be considered the principal vector of Lyme disease in north China. PMID- 14756410 TI - Brevipalpus mites as vectors of unassigned rhabdoviruses in various crops. PMID- 14756411 TI - Brevipalpus californicus, B. obovatus, B. phoenicis, and B. lewisi (Acari: Tenuipalpidae): a review of their biology, feeding injury and economic importance. AB - The genus Brevipalpus includes most of the economically important species of Tenuipalpidae. Many Brevipalpus species reproduce by theletokous parthenogenesis while other species reproduce by male fertilization of female eggs. Previous researchers have determined that Brevipalpus californicus (Banks), B. obovatus Donnadieu, and B. phoenicis (Geijskes) females were haploid with two chromosomes. The life cycle and developmental times for these three species are reviewed. Longevity of each Brevipalpus species is two to three times greater than corresponding longevities of various tetranychid mites. Brevipalpus mites inject toxic saliva into fruits, leaves, stems, twigs, and bud tissues of numerous plants including citrus. Feeding injury symptoms on selected plants include: chlorosis, blistering, bronzing, or necrotic areas on leaves by one or more Brevipalpus mites. Premature leaf drop occurred on 'Robinson' tangerine leaves in Florida (USA). Leaf drop was observed in several sweet orange and grapefruit orchards in Texas (USA) that were heavily infested with Brevipalpus mites feeding on the twigs, leaves, and fruit. Initial circular chlorotic areas appear on both sweet orange and grapefruit varieties in association with developing populations of Brevipalpus mites in Texas. These feeding sites become progressively necrotic, darker in color, and eventually develop into irregular scab-like lesions on affected fruit. Russeting and cracking of the fruits of other plant hosts are reported. Stunting of leaves and the development of Brevipalpus galls on terminal buds were recorded on sour orange, Citrus aurantium L., seedlings heavily infested with B. californicus in an insectary. The most significant threat posed by these mites is as vectors of a potentially invasive viral disease called citrus leprosis. PMID- 14756412 TI - Host plants of Brevipalpus californicus, B. obovatus, and B. phoenicis (Acari: Tenuipalpidae) and their potential involvement in the spread of viral diseases vectored by these mites. AB - The family Tenuipalpidae has over 622 species in 30 genera described worldwide. A total of 928 plant species in 513 genera within 139 families are recorded hosts of one or more of the following species: Brevipalpus californicus (Banks), B. obovatus Donnadieu, and B. phoenicis (Geijskes). B. californicus has 316 plant species reported as hosts compared with 451 and 486 host plants for B. obovatus and B. phoenicis, respectively. There are 67 genera of plants within 33 families that are reported hosts of only B. californicus, 119 genera within 55 plant families that are hosts of only B. obovatus, and 118 genera of plants within 64 families that are hosts of only B. phoenicis. There are 14 genera of plants within 12 families that are hosts to both B. californicus and B. obovatus, while there are 40 genera of host plants within 26 families that are hosts for both B. californicus and B. phoenicis. A total of 70 genera of host plants within 39 families have been reported as hosts of both B. obovatus and B. phoenicis, while 77 genera of plants within 44 families have been reported as hosts of all three Brevipalpus species. Geographical differences in the three species of Brevipalpus identified on different plant species within the same genus are common. PMID- 14756413 TI - Morphological observations on Brevipalpus phoenicis (Acari: Tenuipalpidae) including comparisons with B. californicus and B. obovatus. AB - The genus Brevipalpus has over 300 species worldwide. The three most important agricultural pest species in the genus, Brevipalpus californicus (Banks), B. obovatus Donnadieu, and B. phoenicis (Geijskes), have been consistently confused and misidentified for more than 50 years. The present study provides a discussion of the characters and character states used to separate these mites. Low temperature scanning electron microscopy and traditional light microscopy techniques were used to illustrate the subtle morphological differences between these three species. Morphology of the dorsal propodosoma, opisthosoma, and leg chaetotaxy of all three species was examined and compared. The number of dorsal setae, the number of solenidia (omega) on tarsus II, and dorsal cuticular patterns were the most important characters in the identification of Brevipalpus species. B. phoenicis is similar to B. californicus in having two omega on tarsus leg II and different from B. obovatus which has only one omega on tarsus leg II and similar to B. obovatus in having only one pair of F setae (f3), but differing from B. californicus which has two pairs of F setae (f2-3). The dorsal opisthosomal and propodisomal cuticular patterns frequently used to distinguish between these three species are useful but one must be aware that age, feeding, and mounting techniques can affect the appearance of these characters. PMID- 14756414 TI - Brevipalpus-transmitted plant virus and virus-like diseases: cytopathology and some recent cases. AB - An increasing number of diseases transmitted by Brevipalpus mite species (Acari: Tenuipalpidae) is being identified that affect economically important plants such as citrus, coffee, passion fruit, orchids, and several ornamentals. All of these diseases are characterized by localized lesions (chlorotic, green spots, or ringspots) on leaves, stems, and fruits. Virus or virus-like agents are considered to be the causal agents, possibly transmitted in a circulative propagative manner by Brevipalpus mites. The virus or virus-like particles are short, rod-like, or bacilliform, that induce two characteristic types of cell alteration: (1) 'Nuclear type'--nuclei of parenchyma and epidermal cells in the lesions often contain a large electron lucent inclusion. Short, naked, rod-like (40-50 nm x 100-110 nm) particles may be seen in the viroplasm or nucleoplasm and in the cytoplasm. These particles are commonly arranged perpendicularly on the membranes of the nuclear envelope or endoplasmic reticulum (ER). In a very few instances, they were found to be membrane-bound, within the ER cavities. (2) 'Cytoplasmic type'--short bacilliform particles (60-70 nm x 110-120 nm) are present within the cisternae of the ER and often have electron dense viroplasm of varied shapes present in the cytoplasm. Bacilliform particles may be seen budding into the ER lumen near the viroplasm. These particles resemble those of members of the Rhabdoviridae, but are shorter. The only sequenced virus of this group, orchid fleck virus (OFV), has a negative sense (bipartite) type ssRNA genome, but its organization is similar to known rhabdoviruses, which are monopartite. Both types of cytopathological effects have been found associated with citrus leprosis. In orchids, OFV has a 'nuclear type' of cytopathology, but in some species the 'cytoplasmic type' has been found associated with ringspot symptoms. In Hibiscus and Clerodendron, green spot symptoms have been associated with the cytoplasmic type of cell alteration, while chlorotic spots, in the same species, are associated with the nuclear type. In a few cases, both types of cytopathological effects have been found in the same tissue and cell. PMID- 14756415 TI - Citrus leprosis virus vectored by Brevipalpus phoenicis (Acari: Tenuipalpidae) on citrus in Brazil. AB - Citrus leprosis is caused by Citrus leprosis virus (CiLV) that is transmitted by mites in the genus Brevipalpus (Acari: Tenuipalpidae). This disease directly reduces production and the life span of the citrus plant. The main symptoms of the disease include lesions on fruits, leaves, and twigs or small branches, causing premature fruit drop, defoliation, and death of the twigs or branches leading to serious tree decline. Leprosis is a highly destructive disease of citrus, wherever it occurs. The Brazilian citrus industry spends over 100 million US dollars annually on acaricides to control the vector, Brevipalpus phoenicis (Geijskes). This review contains information about the history of the etiology of citrus leprosis, its geographical distribution, host range, the role of the mite vectors, viral morphology and relationships with the infected cell, and transmissibility of the virus by the mite. In addition, data on the mite-virus plant relationship, disease damage, and strategies for controlling disease spread are presented. PMID- 14756416 TI - Citrus leprosis and its status in Florida and Texas: past and present. AB - According to published reports from 1906 to 1968, leprosis nearly destroyed the Florida citrus industry prior to 1925. This was supported with photographs showing typical leprosis symptoms on citrus leaves, fruit, and twigs. Support for the past occurrence of citrus leprosis in Florida includes: (1) presence of twig lesions in affected orange blocks in addition to lesions on fruits and leaves and corresponding absence of similar lesions on grapefruit; (2) yield reduction and die-back on infected trees; and (3) spread of the disease between 1906 and 1925. Transmission electron microscopy (TEM) examination of tissue samples from leprosis-like injuries to orange and grapefruit leaves from Florida in 1997, and fruits from grapefruit and sweet orange varieties from Texas in 1999 and 2000 did not contain leprosis-like viral particles or viroplasm inclusions. In contrast, leprosis viroplasm inclusions were readily identified by TEM within green non senescent tissues surrounding leprosis lesions in two of every three orange leaf samples and half of the fruit samples obtained from Piracicaba, Brazil. Symptoms of leprosis were not seen in any of the 24,555 orange trees examined across Florida during 2001 and 2002. The authors conclude that citrus leprosis no longer exists in Florida nor occurs in Texas citrus based on: (1) lack of leprosis symptoms on leaves, fruit, and twigs of sweet orange citrus varieties surveyed in Florida: (2) failure to find virus particles or viroplasm inclusion bodies in suspect samples from both Florida and Texas examined by TEM; (3) absence of documented reports by others on the presence of characteristic leprosis symptoms in Florida; (4) lack of its documented occurrence in dooryard trees or abandoned or minimal pesticide citrus orchard sites in Florida. In view of the serious threat to citrus in the U.S., every effort must be taken to quarantine the importation of both citrus and woody ornamental plants that serve as hosts for Brevipalpus phoenicis (Geijskes), B. californicus (Banks), and B. obovatus Donnadieu (Acari: Tenuipalpidae) from countries where citrus leprosis occurs. PMID- 14756417 TI - Coffee ringspot virus vectored by Brevipalpus phoenicis (Acari: Tenuipalpidae) in coffee. AB - Coffee ringspot is characterized by conspicuous ringspot symptoms on leaves, berries, and less frequently on twigs. It is caused by coffee ringspot virus (CoRSV), a short, bacilliform virus (40 nm x 100-110 nm). The virus is not seed borne and is transmitted by Brevipalpus phoenicis (Geijskes). Transovarial transmission within the mite does not occur. CoRSV has been mechanically transmitted to Chenopodium amaranticolor Coste and Reynaud, C. quinoa Wildenow, Beta vulgaris L., and Alternanthera tenella Colla resulting in local lesions. Systemic infection within both C. amaranticolor and C. quinoa occurs. Virions are found in the nucleus or cytoplasm of infected cells, commonly associated with membranes. Occasionally, membrane bounded particles are found within the cisternae of the endoplasmic reticulum. A characteristic electron lucent, nuclear inclusion is commonly found in many infected cells. These cytopathic effects place CoRSV among the nuclear type of Brevipalpus-borne viruses. The disease has been reported in several Brazilian states (Sao Paulo, Parana, Minas Gerais, and Federal District) and recently found in Costa Rica. A similar disease is known in the Philippines, but no information exists about its relationship to CoRSV. Coffee ringspot had no economical significance until recently when a large scale infection was reported in Minas Gerais that resulted in yield loss. PMID- 14756418 TI - Orchid fleck virus: Brevipalpus californicus mite transmission, biological properties and genome structure. AB - Orchid fleck virus (OFV) causes necrotic or chlorotic ring spots and fleck symptoms in many orchid species world-wide. The virus has non-enveloped, bacilliform particles of about 40 nm x 100-150 nm and is sap-transmissible to several plant species. OFV is transmitted by the mite Brevipalpus californicus (Banks) in a persistent manner and efficiently transmitted by both adults and nymphs, but not by larvae. Viruliferous mites retain their infectivity for 3 weeks on a virus-immune host. The genome of OFV consists of two molecules of 6431 (RNA1) and 6001 nucleotides (RNA2). The RNAs have conserved and complementary terminal sequences. RNA1 contains five open reading frames (ORF), and RNA2 encodes a single ORF. Although some of the encoded proteins of OFV have sequences similar to those of proteins of plant rhabdoviruses, OFV differs from viruses in the family Rhabdoviridae in having a bipartite genome. PMID- 14756419 TI - Passion fruit green spot virus vectored by Brevipalpus phoenicis (Acari: Tenuipalpidae) on passion fruit in Brazil. AB - Passion fruit green spot disease was first identified in 1997 after a severe outbreak at Vera Cruz County, state of Sao Paulo, Brazil. Mature yellow fruits of Passiflora edulis Simms f. flavicarpa Degener showed characteristic green spots, 2-5 mm in diameter and patches of green tissues were present on senescent leaves. The devastating effect to passion flower is caused by necrotic lesions that encircle the stems and kill the plant. In severe cases, entire orchards of a few hectares in size have been completely destroyed. The disease was always preceded by heavy infestations of Brevipalpus phoenicis (Geijskes) (Acari: Tenuipalpidae). Transmission electron microscopy of affected tissues (fruits, leaves, and stems) consistently revealed the presence of short, bacilliform particles (50-70 nm x 100-120 nm) in the cisternae of the endoplasmic reticulum, as well as the presence of a dense viroplasm in the cytoplasm. This cytopathic effect has been found in several other Brevipalpus-transmitted or associated viruses and is classified as a cytoplasmic type of disease. Experimental reproduction of the leaf and stem symptoms was achieved by transferring B. phoenicis collected from affected field passion flower plants onto healthy plants. The evidence supports a viral etiology for the disease and the agent was named passion fruit green spot virus. Its relationship with other B. phoenicis related viruses continues to be studied. The disease was also found in the Brazilian states of Bahia, Sergipe, Rondonia, Minas Gerais, Rio de Janeiro, and in the Federal District. Use of one or more of the following acaricides (hexythiazox, fenbutatin-oxide, propargite, quinomethionate, or dicofol) has significantly reduced the incidence of the disease. PMID- 14756420 TI - Human fast skeletal myosin light chain 2 cDNA: isolation, tissue specific expression of the single copy gene, comparative sequence analysis of isoforms and evolutionary relationships. AB - A cDNA clone encoding human fast skeletal myosin regulatory light chain (HSRLC) has been isolated and characterized from a fetal muscle cDNA library. The cDNA contains the coding sequence of 170 amino acids (aa) and 58 and 91 nucleotides in the 5' and 3' untranslated regions (UTRs), respectively. HSRLC is encoded by a single copy gene in the human genome and shows a tissue-specific pattern of expression in skeletal muscle. Comparison of derived amino acid sequence of HSRLC with database sequences reveals highly conserved 12 amino acid residues in a putative calcium-binding region. HSRLC is unique among all RLC sequences in having three consecutive potential phosphorylatable serine residues. The Cys-129 of HSRLC corresponds to the critical Gly-117 of scallop RLC that is essential for its regulatory function. The clusters of hydrophobic residues that are believed to stabilize the binding of NH2-terminal of RLC with myosin heavy chain show high sequence conservation in RLCs. Besides identifying specific targets for functional studies of HSRLC by mutagenesis, the results support the concept of an ancestral gene from which the RLC genes have evolved. PMID- 14756421 TI - Molecular cloning and characterization of a new Na+/H+ antiporter gene from Brassica napus. AB - A new Na+/H+ vacuolar antiporter gene from Brassica napus was cloned. The full length cDNA of B. napus antiporter gene (BnNHX1, GenBank Acc. No. AY189676) was 1819 bp and contained a 1545 bp open reading frame encoding a protein of 515 amino acids. Homology analysis and molecular modeling revealed that BnNHX1 strongly resembled other Na+/H+ antiporter genes. Its encoded protein belonged to a typical sodium/hydrogen exchanger superfamily and shared consensus sequences within predicted transmemberane segments. Southern blot analysis indicated there were multi-copies of BnNHX1 gene present in B. napus genome, which was different from that in Oryza sativa previously reported. Northern blot analysis revealed that BnNHX1 was salt-inducible and its transcript level was most abundant after 24 h treatment with 200 mM sodium chloride. Our studies suggested that BnNHX1 was a new member of the family of recently cloned plant NHX-genes. PMID- 14756422 TI - Conservation of the PRM1 --> PRM2 --> TNP2 domain. AB - In mouse and human, the genes encoding protamines PRM1, PRM2 and transition protein TNP2 are found clustered together on chromosome 16. In addition, these three genes lie in the same orientation to one another and are coordinately expressed in a haploid-specific manner during spermatogenesis. Previously, we have shown that the human PRM1 --> PRM2 --> TNP2 locus exists as a single chromatin domain bounded by two male germ cell-specific MARs, i.e. Matrix Attachment Regions. A third, somatic-specific MAR element lies immediately 3' of the PRM1 --> PRM2 --> TNP2 domain. This MAR maps to a conserved CpG island 5' of the human SOCS-1 gene. Similarly, two candidate MARs flank the mouse Prm1 --> Prm2 --> Tnp2 domain. Comparative analysis of the mouse and human promoter regions identified several conserved regulatory motifs for each of the genes of this cluster. This further establishes the synteny of this region. Global structural similarities and the functional relevance of the associated candidate regulatory elements are discussed. PMID- 14756424 TI - Molecular cloning of a potential Verticillium dahliae resistance gene SlVe1 with multi-site polyadenylation from Solanum licopersicoides. AB - Caused by Verticillium spp. pathogens, verticillium wilt is a common detrimental disease damaging yield production of many important crops. Isolation of verticillium wilt resistance genes and their transgenic application is a fundamental way to control this disease. Here we report the cloning and sequence characterization of a potential Verticillium dahliae Kleb. resistance gene (Ve) from Solanum lycopersicoides Dun. (designated as SlVe1). The nucleotide sequence of SlVe1 is 3400 bp with an ORF of 3156 bp encoding a protein precursor of 1051 amino acids (aa). Unlike tomato Ve1, SlVe1 had a short leader sequence of 22 bp. Multiple polyadenylation sites were detected, which may result from alternative cleavages directed by the common polyadenylation signal AATAAA, and nucleotide sequences of the cleavage sites for polyadenylation conform to PyPyA. Sharing high homologies to tomato verticillium wilt disease resistance genes Ve1 and Ve2, SlVe1 encoded a cell-surface glycoprotein with receptor-mediated endocytosis-like signal. The leucine rich (16.51%) putative SlVe1 protein had a calculated molecular weight of 116.97 kDa with an isoelectric point of 5.22. It possessed a hydrophobic N-terminal signal peptide of 23 aa and 28 predicted significant leucine-rich repeats (LRRs) containing 29 potential N-glycosylation sites (18 being significant). A membrane-associated hydrophobic domain resided within the C terminal, flanked by a neutral/acidic aa rich domain and a neutral/basic aa rich domain. Forty-four predicted phosphorylation sites (28 for S, 5 for T and 11 for Y) distributed in SlVe1, and an endocytosis signal EKWLLW resided in the neutral/basic aa rich C-terminal domain. As compared with Ve1, several clues of variations have been detected in SlVe1 and their possible implications are discussed. PMID- 14756423 TI - Molecular cloning and characterization of a novel human putative transmembrane protein homologous to mouse sideroflexin associated with sideroblastic anemia. AB - Sideroflexin1 (Sfxn1), the prototype of a novel family of evolutionarily conserved proteins present in eukaryotes, has been found mutated in mice with siderocytic anemia. It is speculated that this protein facilitates the transport of a component required for iron utilization into mitochondrial. During the large scale sequencing analysis of a human fetal brain cDNA library, we isolated a cDNA encoding a novel sideroflexin protein (SFXN4), which showed 59% identity and 71% similarity to mouse sideroflexin4. According to the search of the human genome database, SFXN4 gene is mapped to chromosome 10q25-26 and spans more than 24.7kb of the genomic DNA. It is 1428 base pair in length and the putative protein contains 305 amino acids with a conserved predicted five-transmembrane-domains structure. RT-PCR result shows that the SFXN4 gene is expressed in many tissues. PMID- 14756425 TI - Cloning of bovine preproendothelin-2 cDNA and organ distribution of transcripts. AB - Endothelin-2 (ET2), which was originally identified in human, is a bioactive peptide of 21 amino acids with strong vasoconstrictive and pressor effects. Here we report the cDNA cloning and characterization of bovine preproendothelin-2 (PPET2), the precursor form of ET2. The bovine cDNA encodes 177 amino acids of the PPET2 polypeptide, in which a 21-amino acid mature ET2 peptide and a 16-amino acid ET2-like peptide as well as a 23-amino acid putative signal peptide were found. The bovine ET2-like peptide sequence was missing a dibasic amino acid pair at the C-terminal, in contrast to human, mouse and rat, for which the ET2-like sequence is flanked by dibasic pairs at both the N- and C-terminals. Gene expression analysis by RT-PCR showed that the transcript is expressed in various organs including heart, lung, liver, kidney, gastrointestinal tract, uterus and ovary, but not in spleen. Within the gastrointestinal tract, gene expression was detected in rumen, a ruminant-specific digestive organ, as well as stomach, duodenum and colon. PMID- 14756426 TI - Cloning and characterization of human synaptotagmin 10 gene. AB - Synaptotagmin (Syt) is a membrane protein family of secretory vesicles, abundant in neural and some endocrine cells. All the members of this family contain one transmembrane region and two conserved C2 domains. Here we reported a new human Syt 10 gene isolated when screening a human fetal brain cDNA library. This cDNA clone is 3287 bp and contains an open reading frame from 299 to 1870 encoding a putative protein of 523 amino acids. It shares 94.6 and 94.8% homology to rat Syt 10 and mouse Syt 10 at protein level, respectively. RT-PCR result showed that it is expressed only in pancreas, lung and kidney. PMID- 14756427 TI - Clinical research in surgery: the role of the American College of Surgeons Oncology Group. PMID- 14756428 TI - Accelerated partial breast irradiation: a change in treatment paradigm for early stage breast cancer. PMID- 14756429 TI - Randomized trial comparing neo-adjuvant versus adjuvant chemotherapy in operable locally advanced breast cancer (T4b N0-2 M0). AB - BACKGROUND AND OBJECTIVES: Locally advanced breast cancer (LABC) remains a major problem in developing countries. While trials utilizing neo-adjuvant chemotherapy demonstrate superior survival rates compared to historic controls, randomized studies evaluating the precise role of neo-adjuvant chemotherapy in LABC are lacking. In the present trial, neo-adjuvant chemotherapy was compared against adjuvant chemotherapy to assess survival advantage in operable T4b N0-2 M0 breast cancer. METHODS: A total of 101 women with operable LABC (T4b N0-2 M0) were randomized. In arm A, 50 patients received 3 cycles of CEF chemotherapy before and 3 cycles following surgery. In arm B, 51 patients had primary surgery followed by 6 cycles of CEF chemotherapy. In both arms, loco-regional radiotherapy was given after completion of CEF. RESULTS: The response of primary tumor to neo-adjuvant chemotherapy was 66%, complete response (CR) 14% and partial response (PR) 52%. Clinical nodal response occurred in 95% of node positive patients. Only two (4%) patients had pathologic CR both in tumor and axilla. There was a significant (P = 0.02) increase in incidence of pathologically negative nodes in arm A. At a median follow up of 25 months, there was no significant difference in overall and disease free survival (DFS) in both arms (P = 0.42 and 0.18). Patients showing a response to neo-adjuvant chemotherapy had better DFS (P = 0.04) compared to those who had no response. CONCLUSIONS: Early results of the study indicate no survival benefit with the inclusion of neo-adjuvant chemotherapy in LABC (T4b N0-2 M0). Neo-adjuvant chemotherapy resulted in significant down staging; good responders had a better DFS compared to those who did not respond. PMID- 14756430 TI - Large segment allograft survival is improved with intramedullary cement. AB - BACKGROUND AND OBJECTIVES: The most common cause of failure in large segment bone allografts used for oncologic reconstruction is allograft fracture. Reinforcement with pressurized intramedullary cement may improve the mechanical properties of allografts. METHODS: We reviewed the prospectively collected records of 45 patients who underwent en bloc resection of an extremity bone tumor and reconstruction using an allograft with intramedullary cement. RESULTS: Seven allografts were used for arthrodesis, 20 were intercalary, and 18 were osteochondral. Twenty eight of 45 patients were alive at a mean 5.8 years (SD 1.9; range 3-11.2) with 24 allografts in situ. In these 45 patients, there were four allograft fractures and four infections. Six of these complications resulted in allograft removal. The estimated 5-year allograft survival rate was 86% (95% confidence interval 74-98%). Seven patients required secondary autogenous bone grafting for non-unions. Function measured by the Toronto Extremity Salvage Score and the 1987 and 1993 Musculoskeletal Tumor Society Rating Scales demonstrated a consistent pattern with worse function in patients with osteochondral allografts and best function with intercalary allografts. CONCLUSIONS: These results suggest the addition of intramedullary cement to large segment bone allografts improves their survival by decreasing the fracture risk, particularly for allografts used for arthrodesis and intercalary reconstructions. PMID- 14756431 TI - High incidence of urinary bladder involvement in carcinomas of the sigmoid and rectum: a retrospective review of 580 patients with colorectal carcinoma. AB - BACKGROUND AND OBJECTIVES: It is not uncommon for colorectal carcinomas to invade the urinary bladder. However, the actual incidence of urinary bladder invasion of colorectal carcinomas and the common sites of the original tumors are still unclear. METHODS: The clinical records of 580 patients diagnosed with colorectal carcinoma were retrospectively reviewed. A further review of patients with urinary bladder invasion was performed. RESULTS: Of the 580 patients, 17 (2.9%) had a diagnosis of urinary bladder invasion intraoperatively. The incidence of bladder involvement was significantly higher in carcinomas of the sigmoid and rectum than in carcinomas of other colon segments (4.1 vs. 0.5%, P = 0.017). Although a combination of computed tomography (CT) and cystoscopy predicted 80% of the bladder involvement, preoperative diagnostic modalities could not provide information for the differentiation between macroscopic and pathological invasion or the necessity for total cystectomy. CONCLUSIONS: The sigmoid colon and rectum were common sites of the original tumors invading the urinary bladder compared with other colon segments. CT and cystoscopy are recommended for preoperative screening of the bladder involvement of sigmoid colon and rectal cancer. However, it appeared to be difficult to predict the pathological invasion of malignant cells and the necessity for total cystectomy preoperatively. PMID- 14756432 TI - [F-18] fluorodeoxyglucose positron emission tomography as a tool for early recognition of incomplete tumor destruction after radiofrequency ablation for liver metastases. AB - BACKGROUND AND OBJECTIVES: To assess the value of FDG positron emission tomography (PET) for early detection of incomplete tumor destruction after radiofrequency ablation (RFA) for liver metastasis. METHODS: Twenty-eight unresectable liver metastases in 17 patients were treated by RFA. Patients underwent computed tomography (CT) and FDG-PET preoperatively, at 1 week, 1 month, and 3 months postoperatively. Postoperative CT and FDG-PET at 1 week and 1 month were analyzed to identify hypervascular and hypermetabolic residual tumors at the RFA site. These results were correlated with follow-up CT and, in case of reintervention, with pathologic results. RESULTS: In 24/28 of RFA-treated metastases, CT and FDG-PET at 1 week and 1 month showed no tumor residues. During follow-up, none of these 13 patients developed local recurrence at RFA site. In four patients, FDG-PET at 1 week and 1 month showed peripheral hypermetabolic residue after RFA, whereas CT did not revealed residual tumor. In three patients, local persistence of viable tumor cells was biopsy-proven at reintervention. In the fourth, follow-up CT showed subsequent development of a local recurrence. CONCLUSIONS: FDG-PET accurately monitors the local efficacy of RFA for treatment of liver metastases, as it early recognizes incomplete tumor ablation, not detectable on CT. PMID- 14756433 TI - Prognostic value of intratumoral CD8+ T lymphocyte in extrahepatic bile duct carcinoma as essential immune response. AB - BACKGROUND AND OBJECTIVES: Tumor-infiltrating lymphocytes form an important aspect of the host defense against an expanding neoplasm. CD8+ T cells have been identified as a prognostic factor in several cancers. Here, we investigate that the influence of CD8+ T cells on extrahepatic bile duct carcinoma (EBDC) patient survival. METHODS: CD8+ T cell immunoreactivity in 58 surgically resected EBDC specimens was investigated. The relationship between CD8+ T cell immunoreactivity and clinical and histopathologic features was analyzed. RESULTS: Thirty-two tumors (55%) possessed intratumoral CD8+ T cells. The degree of intratumoral CD8+ T cell immunoreactivity demonstrated a significant relationship to lower numbers of lymph node metastasis, reduced venous invasion, decreased perineural invasion, and better pTNM staging. Intratumoral CD8- T cells were also associated with increased patient survival. Multivariate analysis indicated that the presence of intratumoral CD8+ T cells was an independent prognostic factors. CONCLUSIONS: The infiltration of a cancer cell nest by CD8+ T cells is a reliable marker predicting increased survival of patients with EBDC. PMID- 14756434 TI - Malignant melanoma developing in an area of hereditary palmoplantar keratoderma (Mal de Meleda). AB - Palmoplantar keratoderma (PPK) refers to a genetically heterogeneous group of skin diseases, which may be inherited in autosomal dominant or recessive fashion. We observed a case of a 74-year-old man with Mal de Meleda, who developed malignant melanoma inside the hyperkeratotic palmar skin of the right hand. Many authors have reported a higher incidence of cancer in cases affected by palmoplantar hyperkeratosis both for hereditary association and particularly for mechanical damage of the affected areas. The association with melanoma has already been described, but not in a true Mal de Meleda type syndrome as in the case reported in this paper. PMID- 14756435 TI - The evolution of lung cancer screening. AB - In the 1970s, four trials failed to demonstrate any mortality reduction using a combination of chest X-ray (CXR) and/or sputum cytology. The recent early lung cancer action project (ELCAP) demonstrated that modern screening is capable of detecting Stage I lung cancers. Bronchial epithelial changes leading up to cancers are now being understood to include histologic changes and genetic alterations. Emerging molecular markers detected in sputum and serum show promise in the future of lung cancer screening. PMID- 14756436 TI - Tumor markers and colorectal cancer: utility in management. AB - Colorectal cancer (CRC) is a leading cause of morbidity and mortality. Although genetic testing can screen for rare hereditary CRC syndromes, there is no ideal means of screening for sporadic forms of CRC. This review will focus on markers that are currently used in the management of sporadic CRC and their limitations, as well as possible future clinical applications. PMID- 14756437 TI - A new proposal of skin-closure system for median sternotomy: usefulness and cosmetic results analysis of MEDIZIP Surgical Zipper in neoplastic immuno compromised patients. AB - BACKGROUND AND OBJECTIVES: The authors evaluate the effectiveness and the cosmetic results obtained using the new skin closing system MEDIZIP Surgical Zipper in oncological immuno-compromised patients submitted to median sternotomy. METHODS: In our Institute, from 1999 to 2002, MEDIZIP was used to close the sternal wounds in 45 patients undergoing median sternotomy for bilateral metastasectomy, It took about half a minute (mean time: 32.00 +/- 11.48 sec) to perform the application. To evaluate the cosmetic results, a three-level scale was conceived: level 1: very good, level 2: satisfactory, level 3: inadequate. RESULTS: Overall forty-two 20 cm-long zippers were used, two 25 cm-long and one 30 cm-long. MEDIZIP remained in situ for an average of 9.98 +/- 2.23 days (median: 9 days; range: 8-13 days). The average time taken for inspection was 70.00 +/- 2.48 sec (median: 70, range: 45-130) and the zipper was removed in a few seconds. No wound infections were observed. We classified 39 patients at level 1 (very good, 87%), and 6 at level 2 (satisfactory, 13%). CONCLUSIONS: MEDIZIP can be considered an effective skin-closure system which is easily and quickly handled and assures good cosmetic results with non-invasive removal; it proves particularly useful in pediatric patients and in adults affected by neoplastic diseases and undergoing multiple combined anti-cancer treatments. PMID- 14756438 TI - Sub-mammary approach for pectoralis major myocutaneous flap in females. PMID- 14756439 TI - Matrix metalloproteinase-2 and -9 expression in astrocytic tumors. AB - In this study, we investigated whether the expression of matrix metalloproteinase (MMP)-2 and MMP-9 correlated with invasiveness, proliferative potential, or prognosis in astrocytic tumors. Thirty-seven astrocytic tumors (8 diffuse astrocytomas, 15 anaplastic astrocytomas, and 14 glioblastomas) and three gliomatosis cerebri were investigated immunohistochemically. The invasive glioma group included three cases of gliomatosis cerebri and two of glioblastoma associated with cerebrospinal fluid dissemination. The expression of MMP-2 and MMP-9 was evaluated by assigning an immunohistochemical (IHC) score defined as the sum of expression frequency and intensity. mRNA expression patterns for the MMPs were also evaluated in a reverse transcription-polymerase chain reaction assay. Neither the MMP-2 nor MMP-9 IHC score was related to histological malignancy. The MMP-2 IHC score of the invasive glioma group was significantly higher than those of other kinds of astrocytic tumors. However, the MMP-9 IHC score did not correlate with dissemination among astrocytic tumors. An inverse correlation was observed between the MIB-1 labeling index and the IHC scores of MMP-2, but it was not significant. A Kaplan-Meyer survival analysis revealed no significant relationship between the survival rate and MMP-2 or MMP-9 expression. Our study showed that MMP-2 expression, but not MMP-9 expression, may be associated with invasion in astrocytic tumors. PMID- 14756440 TI - A fourth ventricle atypical teratoid/rhabdoid tumor in an infant. AB - Atypical teratoid/rhabdoid tumor (AT/RT), a recently established central nervous system tumor entity, occurs in children and is more malignant than medulloblastoma/primitive neuroectodermal tumors (PNET). We report here a case of AT/RT in a male infant who was 9 months old at the time of diagnosis. Magnetic resonance imaging revealed that the tumor occupied the fourth ventricle, and at surgery it was found to adhere to the floor of the fourth ventricle. After subtotal removal of the tumor mass, chemotherapy and radiotherapy were performed, but the patient died about 8 months after the diagnosis following rapid regrowth of the residual tumor. Light-microscopically, the tumor was composed mainly of nests of rhabdoid cells with fields of PNET. Occasional mesenchymal and epithelial fields were also evident. Immunohistochemically, these rhabdoid cells were positive for vimentin, epithelial membrane antigen, smooth-muscle actin, cytokeratin, and S-100 protein, and less frequently for glial fibrillary acidic protein. Electron-microscopically, the typical rhabdoid cells contained whorled bundles of intermediate filaments in their cytoplasm. Occasionally, such rhabdoid cells were covered partially by basal lamina at their stromal interface. These findings are typical of AT/RT. Although it is well known that AT/RT often arises in the posterior fossa, detailed reports of cases affecting the fourth ventricle are rare. In this case, the ultrastructural relationship between rhabdoid cells and the basal lamina, which has not so far been described in AT/RT, was of great interest when the nature of the rhabdoid cells was considered. PMID- 14756441 TI - Expression of fascin, an actin-bundling protein, in astrocytomas of varying grades. AB - Malignant astrocytomas are highly infiltrative neoplasms that invade readily into regions of normal brain. On a cellular basis, the motility and invasiveness of human cancers can be ascribed in part to complex rearrangements of the actin cytoskeleton that are governed by several actinbinding proteins. One such actin binding protein that has been linked to the invasive behavior of carcinomas is fascin, which serves to aggregate F actin into bundles. In this study, we examined the expression of fascin in a series of human malignant astrocytomas (WHO grades I-IV). Five grade I, 5 grade II, 10 grade III, and 26 grade IV human astrocytomas were examined for fascin and glial fibrillary acidic protein (GFAP) expression by double immunofluorescence confocal microscopy. Expression of fascin and GFAP was also determined by Western blot analysis. Fascin expression increased with increasing WHO grade of astrocytoma. This is in marked contrast to GFAP expression, which decreased with increasing WHO grade. In grades I and II neoplasms, and within non-neoplastic brain, fascin and GFAP were expressed diffusely within regions examined. However, in the higher-grade astrocytomas (grades III and IV), fascin and GFAP were expressed regionally in distinctly separate tumor cell populations. This is the first study to demonstrate the expression of fascin in human astrocytic neoplasms. The role that fascin plays in contributing to the invasive phenotype of anaplastic astrocytomas awaits further study and investigation. PMID- 14756442 TI - Detection of gene amplification and deletion in high-grade gliomas using a genome DNA microarray (GenoSensor Array 300). AB - Glioblastoma is a rapidly growing tumor that accounts for more than 50% of all primary gliomas. Amplification of oncogenes and deletion of tumor suppressor genes frequently affects tumor progression. Thus, the goal of this study was to conduct a comprehensive analysis of gene aberrations of individual glioblastomas. A genome DNA microarray (GenoSensor Array 300), spotted with 287 target genes, was used to analyze resected tissue from 11 different high-grade gliomas. The average number of gene aberrations was 9.0 per case (WHO grade III) and 13.3 per case (WHO grade IV). EGFR was the most frequent amplified gene in this series (4 of 11 cases), and high-level amplification was also detected for EGFR, SAS/CDK4, and AKT1. A high frequency of deleted genes was observed in 6 of 11 cases (54.5%), including FGFR2, MTAP, and DMBT1. The detected gene aberrations were matched to the classical primary glioblastoma pathway in five of nine cases. We conclude that the GenoSensor Array 300 genomic DNA microarray is a useful method for the comprehensive identification of amplified and deleted genes in glioblastoma. PMID- 14756443 TI - The expression of matrix metalloproteinase-2 and -9 in human gliomas of different pathological grades. AB - Matrix metalloproteinases (MMPs) have been implicated to play a critical role in glioma invasiveness. In this study, we aimed to investigate the expression of MMP 2 and MMP-9 in human gliomas of different degrees of malignancy, and evaluated the correlation between MMP-2 and MMP-9 expression in gliomas. The samples from 65 cases of glioma were divided into four groups according to the WHO classification: there were 16 cases of grade I, 17 cases of grade II, 20 cases of grade III, and 12 cases of grade IV. Normal brain samples served as the control group, and biopsy specimens were obtained from 8 glioma patients with a needle placed into the adjacent brain 1 cm from the margin after tumor resection. All the samples were stained with hematoxylin and eosin and immunohistochemistry. A computer-aided image-analysis system was employed to measure the integral optical density (IOD) of positive slides. No positive staining was found in the control group. The positive staining was localized in the cytoplasm of glioma cells, the extracellular matrix (ECM), the basement membrane (BM), and the endothelial cells of blood vessels. Positive staining rates increased significantly when the degree of malignancy of gliomas was elevated. The IOD value of MMP-2 and MMP-9 also indicated that the intensity of MMP-2 and MMP-9 expression was elevated significantly with the degree of malignancy of the gliomas. There was a positive correlation between MMP-2 and MMP-9 expression in gliomas. Glioma invasion and angiogenesis were particularly seen in the biopsied tissues, and MMP-9 immunostaining seemed to be much more intense and extensive than MMP-2 immunostaining in these samples. These results suggest that MMP-2 and MMP-9 staining in gliomas is localized in the cytoplasm of tumor cells, BM, and endothelial cells, and that MMP-2 and MMP-9 together play an important role in the invasiveness of gliomas, mediating the degradation of the ECM and angiogenesis. MMP-2 and MMP-9 could be molecular targets in the treatment of malignant glioma. PMID- 14756444 TI - Immunohistochemical analysis of the p53 family members in human craniopharyngiomas. AB - Craniopharyngiomas are intracranial tumors that usually arise in the site around the sella turcica. They are composed of distinctive sheets of epithelial cells showing adamantinomatous or squamous-papillary histologic types. Because little is known about the tumorigenesis of craniopharyngiomas, we retrieved samples from 15 tumor cases to investigate the functional significance of the p53 family of transcription factors, which are known to be expressed in various human epithelia. Immunohistochemical analysis of these cases demonstrated similar expression profiles of p53 family members in the two histologic types of the tumor; i.e., strong nuclear expression of p63 was observed in all cell layers, and moderate to intense nuclear expression of p73 was observed in the basal cell layers. In contrast to p63 and p73, the reactivity of an archetypal tumor suppressor, p53, was occasional and weak in the two histologic types. Because p63 was widely expressed in the tumors, reverse transcription-polymerase chain reaction (RT-PCR) analysis was conducted to elucidate which spliced variant of p63 was expressed. The results showed that deltaNp63, lacking a terminal transactivation domain of p63, was the dominant isoform. Together with the reported evidence that the deltaNp63 isoform is highly expressed in human squamous-cell carcinomas, these data suggest that the cellular architecture characteristic of the expression of p53 family members may be required for the histogenesis of craniopharyngiomas, where deltaNp63 has a possible role in maintaining proliferative activity of the tumor cells, like squamous-cell carcinomas in other tissues. PMID- 14756445 TI - A rare case of cellular schwannoma involving the trigeminal ganglion. AB - Cellular schwannomas rarely involve the cranial nerves, being more common in the spinal and peripheral nerves. A rare case of cellular schwannoma involving the gasserian ganglion, a hitherto unreported site, that extended infratentorially to present as a cerebellopontine angle tumor is reported. It is important to recognize that cellular schwannomas can histologically mimic malignant peripheral nerve sheath tumors because of their high cellularity and mitotic activity, but they are relatively benign tumors with a tendency to recur but not metastasize. PMID- 14756446 TI - Parenchymal anaplastic ependymoma with intratumoral hemorrhage: a case report. AB - We report an unusual case of a 56-year-old woman with a supratentorial anaplastic ependymoma localized in the parenchyma without continuity with the ventricular system and brain surface. The patient presented with vertigo, and a calcified mass was detected in the left temporal parenchyma. Five years later, she had seizure of the right extremities. Computed tomographic scanning and magnetic resonance imaging revealed an enhanced mass with an intratumoral hemorrhage adjacent to the calcified mass. Subtotal removal of the tumor was performed. The histological analysis revealed that the tumor was an anaplastic ependymoma. After focal radiation therapy (50 Gy), the outcome was favorable, although the residual lesion was still seen on the images. Ependymomas usually arise from the cells lining the ventricular system and the central canal of the spinal cord. We discuss the summary of published cases of supratentorial ectopic ependymoma since the first case in 1995. PMID- 14756447 TI - Salmon roe-like amyloid deposition in a prolactinoma: a case report. AB - An unusual case of prolactin-producing adenoma with extensive amyloid deposition is reported to clarify its radiological, intraoperative, and light- and electron microscopic findings. A 41-year-old female patient complained of amenorrhea persisting for 20 years. Magnetic resonance imaging (MRI) revealed a pituitary adenoma, which included low-intensity spots on T1- and T2-weighted images. Intraoperative examination found multiple small, yellowish, spherical masses resembling salmon roe within the adenoma. Light and electron microscopy revealed the presence of immunoreactive cells for prolactin intermingled with concentric lamellar bodies of radially arranged amyloid fibrils originating from the endoplasmic reticulum in prolactinoma cells. The extracellular lamellar amyloid deposits were apparently due to degradation of prolactin-producing cells, but the reason for the production and radially arranged accumulation of amyloid remains to be identified. PMID- 14756448 TI - Gliomatosis cerebri in a young patient showing various cranial nerve manifestations: a case report. AB - A case of gliomatosis cerebri in a 27-year-old man showing various cranial nerve manifestations is described. He was diagnosed as having cranial mononeuritis multiplex (bilateral oculomotor nerve paralysis, left facial nerve paralysis, bulbar palsy manifestations, and hypoglossal nerve paralysis) and was hospitalized in the neurology department on August 1, 2000. Although he continued to visit the neurology department after discharge, his manifestations showed no improvement. He was sent to our department for brain biopsy in August 2001. A biopsy performed at the Sylvian fissure from the frontal lobe/temporal lobe cortex showed high intensity on T2-weighted and Flair magnetic resonance imaging (MRI). The pathological findings were diffuse low-grade astrocytoma infiltrating between the pia mater and the cerebral cortex. We believed that the astrocytoma spreading on the subpia mater was responsible for the various cranial nerve manifestations, and we started whole-brain irradiation (46 Gy) + interferon (IFN) beta D.I.V. from September 2001. The pathological findings of the brain biopsy showed diffuse astrocytoma. The clinical presentation was dramatically improved after radiotherapy. It seemed that this tumor had spread along the subpia mater and subependyma. When he was discharged in early December, he walked by himself. The characteristic features of this case are that no lesion in the cerebellum or brain stem was found on MRI, even though the main manifestations were cerebello brain stem manifestations, and biopsy of the cerebral cortex revealed astrocytoma. It should be noted that the clinical manifestations of astrocytoma in some cases are dissociated from the imaging observations. PMID- 14756449 TI - Arsenic trioxide induces selective tumour vascular damage via oxidative stress and increases thermosensitivity of tumours. AB - It has previously been found that the anti-leukaemia agent Arsenic Trioxide (ATO) causes vascular shutdown in solid tumours and markedly sensitizes tumours to hyperthermia. The present study was designed to evaluate the mechanism of action and dose-dependence of ATO-induced thermosensitization in FSaII and SCK murine tumours. The role of oxidative stress was studied by observing ATO-induced vascular shutdown in vivo and ATO-induced endothelial cell adhesion molecule expression in vitro in the presence or absence of an anti-oxidant. It was found that a dose as low as 2 mg/kg ATO impaired vascular function, as estimated by 86Rb uptake, in the tumour. The degree of tumour growth delay induced by 1 h of hyperthermia at 42.5 degrees C, applied 2 h after ATO injection, was proportional to the dose of ATO administered. In addition, it was found that ATO can directly thermosensitize tumour cells in vitro. The development of massive tissue necrosis in the tumour was observed in the days after treatment, especially with the combination of ATO and heating. ATO-induced adhesion molecule expression in vitro was abolished when the anti-oxidant n-acetyl-cysteine (NAC) was introduced prior to exposure, while the addition of NAC in vivo partially blocked ATO-induced vascular shutdown. These results suggest that the expression of adhesion molecules by the vasculature due to oxidative stress contribute to the ATO induced selective tumour vascular effects observed and that the clinical use of ATO to increase tumour thermosensitivity via direct cellular and vascular effects appears feasible. PMID- 14756450 TI - Heat-induced growth inhibition and apoptosis in transplanted human head and neck squamous cell carcinomas with different status of p53. AB - To examine p53-dependency in hyperthermic cancer therapy, heat-induced growth inhibition and apoptosis in transplanted human head and neck squamous cell carcinoma (HNSCC) tumours were analysed with different status of p53 into nude mice. The tumour tissue from HNSCC cell line (SAS) transfected with mutant p53 gene (SAS/mp53) or control vector containing neo gene (SAS/neo) was transplanted into the subcutaneous tissue of the thigh of nude mice using a trocar. Hyperthermia was performed at 42 degrees C when the mean diameter of tumour was 5 6mm. The diameter of tumours was measured using vernier calipers and tumour weight (TW) and the relative tumour weight (RW) was calculated. Tumour regrowth delay was evaluated when the RW reached 5-fold against the control group. The accumulation of p53 and Bax proteins was examined by an immunohistochemical technique. Apoptotic cells in the sections were detected by staining of DNA ends using an immunohistochemical technique. SAS/mp53 tumours showed more heat resistance than SAS/neo tumours. The p53-positively staining cells were observed in untreated SAS/mp53 tumours, but not in untreated SAS/neo tumours. After heat treatment, the accumulation of p53 and Bax proteins was observed in SAS/neo tumours, but little in SAS/mp53 tumours. The incidence of apoptotic cells induced by heat treatment was very low in SAS/mp53 tumours compared with SAS/neo tumours. In conclusion, the heat-induced growth inhibition of a transplanted HNSCC may be correlated with the induction of p53-dependent Bax-mediated apoptosis. Thus, p53 status appears to be one of the useful parameters for the predictive assays in hyperthermic cancer therapy. PMID- 14756451 TI - Improvement of absorbing structures used in regional hyperthermia. AB - Local pain is a major limiting factor in regional hyperthermia treatment with radiative applicators. Absorbing structures, consisting of agar bound saline water, have been used successfully to reduce peripheral hot spots. However, both clinical experience and simulation results indicate a SAR elevation in the tissue under the edges of the absorber block. This paper investigates the effect of modification of shape, position and spatial composition of the absorber blocks on the central attenuating effect and the SAR elevating effect at the edges. A selection from a set of five options is made based on simulations with a phantom and a single ring dipole applicator. The simulations have been performed with the FDTD core of the regional hyperthermia treatment planning system. It is shown that tapering of the absorber edge and introduction of a water layer between the absorber and the skin can reduce the edge effect in the superficial fat layer by approximately 50% with respect to a rectangular absorber. A further reduction of 15% can be obtained by an absorber with an appropriate gradient of its conductivity in the direction of the dominant E-field. The modified absorbers produce a central attenuating effect comparable to the rectangular type. The use of a water layer type and a sigma gradient type absorber is also analysed in a patient anatomy, both in the dipole ring applicator, operating at 70 MHz, as well as in a three ring Cavity Slot (CS) applicator, operating at 150 MHz. The mutual influence of phase-amplitude steering and the application of absorbers is investigated in the CS applicator. It appears that absorbers have a significant influence on the interference pattern in the patient model, possibly causing substantial reduction of the SAR value in the tumour and limiting the possibility of ad hoc application of absorbers. Re-optimization can only partly cancel this effect. Local SAR reduction by phase-amplitude control alone can match or improve the effect obtained with modified absorbers. PMID- 14756452 TI - Microwave thermal imaging: initial in vivo experience with a single heating zone. AB - The deployment of hyperthermia as a routine adjuvant to radiation or chemotherapy is limited largely by the inability to devise treatment plans which can be monitored through temperature distribution feedback during therapy. A non invasive microwave tomographic thermal imaging system is currently being developed which has previously exhibited excellent correlation between the recovered electrical conductivity of a heated zone and its actual temperature change during phantom studies. To extend the validation of this approach in vivo, the imaging system has been re-configured for small animal experiments to operate within the bore of a CT scanner for anatomical and thermometry registration. A series of 5-7 day old pigs have been imaged during hyperthermia with a monopole antenna array submerged in a saline tank where a small plastic tube surgically inserted the length of the abdomen has been used to create a zone of heated saline at pre-selected temperatures. Tomographic microwave data over the frequency range of 300-1000 MHz of the pig abdomen in the plane perpendicular to the torso is collected at regular intervals after the tube saline temperatures have settled to the desired settings. Images are reconstructed over a range of operating frequencies. The tube location is clearly visible and the recovered saline conductivity varies linearly with the controlled temperature values. Difference images utilizing the baseline state prior to heating reinforces the linear relationship between temperature and imaged saline conductivity. Demonstration of in vivo temperature recovery and correlation with an independent monitoring device is an important milestone prior to clinical integration of this non-invasive imaging system with a thermal therapy device. PMID- 14756453 TI - Characterization of the SAR-distribution of the Sigma-60 applicator for regional hyperthermia using a Schottky diode sheet. AB - INTRODUCTION: Characterization of the performance of an hyperthermia applicator by phantom experiments is an essential aspect of quality assurance in hyperthermia. The objective of this study was to quantitatively characterize the energy distribution of the Sigma-60 applicator of the BSD2000 phased array system operated within the normal frequency range of 70-120 MHz. Additionally, the accuracy of the flexible Schottky diode sheet to measure E-field distributions was assessed. MATERIAL AND METHODS: The flexible Schottky diode sheet (SDS) consists of 64 diodes mounted on a flexible 125 microm thick polyester foil. The diodes are connected through high resistive wires to the electronic readout system. With the SDS E-field distributions were measured with a resolution of 2.5 x 2.5 cm in a cylindrical phantom, diameter of 26 cm and filled with saline water (2 g/l). The phantom was positioned symmetrically in the Sigma-60 applicator. RF power was applied to the 4-channel applicator with increasing steps from 25W to a total output of 400 W. RESULTS: The complete system to measure the E-field distribution worked fine and reliably within the Sigma-60 applicator. The E-field distributions measured showed that the longitudinal length of the E-field distribution is more or less constant, e.g. 21-19 cm, over the frequency range of 70-120 MHz, respectively. As expected, the radial E-field distributions show a better focusing towards the centre of the phantom for higher frequencies, e.g. from 15.3-8.7 cm diameter for 70-120 MHz, respectively. The focusing target could be moved accurately from the left to the right side of the phantom. Further it was found that the sensitivity variation of nine diodes located at the centre of the phantom was very small, e.g. < 3% over the whole frequency range. CONCLUSION: The SAR distributions of the Sigma-60 applicator are in good agreement with theoretically expected values. The flexible Schottky diode sheet proves to be an excellent tool to make accurate, quantitative measurements of E-field distributions at low (25 W) and medium (400 W) power levels. An important feature of the SDS is that it enables one to significantly improve quantitative quality assurance procedures and to start quantitative comparisons of the performance of the different deep hyperthermia systems used by the various hyperthermia groups. PMID- 14756454 TI - Increasing the systemic temperature during regional hyperthermia: effect of a cooling strategy on tumour temperatures and side-effects. AB - In the application of regional hyperthermia, optimization of the temperature distribution remains necessary. One of the tools that might be used is a modest increase in the systemic temperature to diminish cooling by blood perfusion. This study investigates (1) if it is feasible to increase the systemic temperature by applying other cooling strategies, without inducing unacceptable systemic stress, and (2) whether a rise in systemic temperature results in improvement of tumour temperatures. Eleven patients with locally advanced cervical carcinoma and 12 patients with locally advanced prostate carcinoma were treated with our Coaxial TEM regional hyperthermia system. In this system, the temperature of the open water bolus can be easily adjusted. Two cooling methods were applied alternately, one with a relatively low water temperature (method A), the other with a higher water bolus temperature in combination with extensive head/chest cooling by a hand shower (method B). Method B resulted in significantly higher systemic temperatures, for both patient groups separately (0.8, respectively, 0.5 degrees C) and for the total patient group (0.7 degrees C). Additionally, all tumour index temperatures were higher. For the combined group (for T50: 0.4 degrees C) and for the cervix group (for T50: 0.7 degrees C), it reached statistical significance. The raise in core temperature led to a significantly higher increase in heart rate. For the group of cervix patients, higher systemic temperatures resulted in more treatment-limiting systemic stress. For the prostate patients, systemic stress was not an important issue. Since the raise in systemic temperature did not influence the overall tolerance of treatment, method B could be applied to this group. However, the increases in tumour temperatures were small, and potential hazards of systemic temperature increase should be considered. PMID- 14756455 TI - Russian olive. PMID- 14756456 TI - Is oral immunotherapy too good to be true? PMID- 14756457 TI - The epidemiology and natural history of asthma: Outcomes and Treatment Regimens (TENOR) study. PMID- 14756458 TI - Observational studies of drug effectiveness. PMID- 14756459 TI - The yellow zone in asthma treatment: is it a gray zone? AB - OBJECTIVE: To review the available literature on methods of preventing and minimizing exacerbations and to target problems for improvement. DATA SOURCES: PubMed and Cochrane Review searches of the English-language literature using the following key yellow zone terms: asthma exacerbation, self-management plans, inhaled corticosteroids, and acute management of asthma exacerbations. STUDY SELECTION: Articles relevant to our yellow zone intervention inquiry. RESULTS: The National Institutes of Health (NIH) guidelines advocate that physicians give patients written action plans with instructions on managing home (yellow zone) exacerbations. However, the criteria used to identify the yellow zone are ambiguous and often confusing to patients and physicians. In addition, apart from a passing mention that doubling doses of inhaled corticosteroids may be an option in asthma step-up care, the guidelines contain no recommendations for yellow zone treatment strategies. This deficiency is directly related to the paucity of organized evidence on the efficacy of the various pharmacological interventions that can be used during an exacerbation. CONCLUSIONS: Translating the NIH guidelines into realistic clinical practice requires a clearer and more patient friendly definition of the yellow zone, and this improved definition will facilitate the prescription of effective interventions in the management of yellow zone exacerbations. PMID- 14756460 TI - Progressive muscle weakness in a 4-year-old girl. PMID- 14756461 TI - Clinical efficacy of microencapsulated timothy grass pollen extract in grass allergic individuals. AB - BACKGROUND: Conventional allergen immunotherapy is clinically effective in reducing the symptoms of allergic rhinitis and asthma. It differs from other pharmacotherapies in that it can induce long-term clinical remission of these diseases. However, it requires years of treatment and is associated with serious allergic reactions. OBJECTIVE: To evaluate the safety, clinical efficacy, and immunologic mechanisms of immunotherapy with an oral, microencapsulated form of timothy grass allergen. METHODS: In this double-blind, placebo-controlled study, 24 patients aged 19 to 55 years with grass pollen allergy were randomized to receive either microencapsulated timothy grass pollen extract or placebo once a day for 10 weeks. The dose of study drug was doubled weekly. Safety was evaluated through weekly visits, daily symptom diaries, and routine laboratory tests. Efficacy was evaluated by comparing medication use and symptoms scores during peak grass pollen season before and after treatment. Allergen-specific T-cell responses, cytokine production, and IgG, IgE, and skin reactivity were measured to evaluate immunologic mechanisms. RESULTS: Eleven of 12 patients in the active treatment group had a decrease in the combined medication and symptom score, but only 4 of 10 patients in the placebo group had a decrease in scores. The proliferative response to timothy grass was reduced by at least 30% in 9 of the 12 grass-treated patients, but only 3 of 11 placebo patients had a proliferative response reduction. Timothy grass-induced interleukin-5 messenger RNA was reduced in the active group, but not in the placebo group. There were no significant changes in either group in IgG, IgE, and skin reactivity. CONCLUSIONS: Oral immunotherapy with microencapsulated allergen induces a form of immunologic tolerance to the allergen and is a safe, efficient, and effective method of allergen immunotherapy. PMID- 14756463 TI - Inhaled anti-inflammatory pharmacotherapy and subsequent hospitalizations and emergency department visits among patients with asthma in the Texas Medicaid program. AB - BACKGROUND: Rates of asthma-related hospitalizations and emergency department (ED) visits continue to rise in the United States. The National Asthma Education and Prevention Program recommends the use of controller pharmacotherapy for patients with persistent asthma. OBJECTIVE: To investigate the influence of initiating inhaled anti-inflammatory (IAI) pharmacotherapy following an asthma related hospitalization or ED visit on risk of subsequent morbid events. METHODS: Texas Medicaid asthma-related medication and medical services claims for September 1997 to July 2001 were extracted. An asthma-related morbid event served as the index event (ED visit or hospitalization for cohort 1; hospitalization for cohort 2). Members of both cohorts were then followed up until a subsequent morbid event occurred or until 1 year after index. Logistic regression was used to compare patients who used IAI medication within 100 days following their index event with nonusers. RESULTS: Controlling for demographic and resource use variables, there was a 52% reduction in the risk of a subsequent ED visit or hospitalization in the year following the index event among users of IAI medication within cohort 1 (risk ratio [RR], 0.485; 95% confidence interval [CI], 0.416-0.565; P < .001). There was a 61% reduction in the risk of a subsequent hospitalization among users of IAI medication within cohort 2 (RR, 0.393; 95% CI, 0.284-0.545; P < .001). CONCLUSIONS: Less than half of the patients had a prescription claim for an IAI medication within 100 days following their index event. Patients who received these medications had a lower risk of a subsequent asthma-related morbid event for the next year. PMID- 14756462 TI - Design and baseline characteristics of the epidemiology and natural history of asthma: Outcomes and Treatment Regimens (TENOR) study: a large cohort of patients with severe or difficult-to-treat asthma. AB - BACKGROUND: Patients with severe and difficult-to-treat asthma represent a small percentage of asthma patients, yet they account for much of the morbidity, mortality, and cost of disease. The goal of The Epidemiology and Natural History of Asthma: Outcomes and Treatment Regimens (TENOR) study is to better understand the natural history of asthma in these patients. OBJECTIVE: To describe the methods and baseline characteristics of the TENOR study cohort. METHODS: The TENOR study is a 3-year, multicenter, observational study of patients with severe or difficult-to-treat asthma. From January through October 2001, more than 400 US pulmonologists and allergists enrolled patients. Patients 6 years or older who were considered to have severe or difficult-to-treat asthma by their physicians were eligible. Patients have been receiving care for 1 year or more, have a smoking history of 30 pack-years or less, and have current high medication or health care utilization in the past year. Data are collected semiannually. RESULTS: A total of 4,756 patients enrolled and completed a baseline visit. Overall, 73% of the TENOR study patients are adults, 10% are adolescents, and 16% are children. According to physician evaluation, 48% of patients have severe asthma, 48% have moderate asthma, 3% have mild asthma, and 96% have difficult-to treat asthma. Severe asthmatic patients have the highest health care utilization in the past 3 months (P < .001). CONCLUSIONS: The TENOR study is the largest cohort of patients with severe or difficult-to-treat asthma. Although patients are equally divided into moderate or severe asthma categories, most are considered difficult-to-treat. The TENOR study highlights the lack of control in moderate-to-severe asthma and provides a unique opportunity to examine factors related to health outcomes in this understudied population. PMID- 14756464 TI - Validity and responsiveness of a brief, asthma-specific quality-of-life instrument in children with acute asthma. AB - OBJECTIVE: To test the validity and short-term responsiveness to change of a pediatric, asthma-specific, health-related quality-of-life (HRQL) instrument. METHODS: Children 2 years and older treated in the emergency department (ED) for acute asthma were eligible for this prospective cohort study. A 10-item instrument, the Integrated Therapeutics Group Child Asthma Short Form (ITG-CASF), was administered at the time of the ED visit and again 14 days later (via telephone). At the follow-up call, parents were also asked about the child's current overall asthma status, missed school or limited activities, and persistence of asthma symptoms. RESULTS: A total of 121 children were enrolled (mean age, 7.9 years), and follow-up was complete for 96 (79%). Mean +/- SD ITG CASF scores at follow-up were significantly higher among children reported to have improved overall (61.8 +/- 19.6) than those not improved (41.9 +/- 21.2), and there was a significant correlation between ITG-CASF score at follow-up and the number of days of school missed or limited activities (r = -0.45; 95% confidence interval [CI], -0.24 to -0.66). There was also a significant difference in improvement in ITG-CASF score from ED visit to follow-up among those improved (13.7-point improvement) compared with those not improved (3.3 point improvement; difference = 10.4; 95% CI, 1.2 to 19.5). The effect size was 0.68, indicating a large responsiveness to change. CONCLUSIONS: The ITG-CASF is a valid and responsive measure of HRQL in children with acute asthma and may be a useful outcome measure in evaluating ED treatment. PMID- 14756465 TI - Differences in respiratory symptoms and pulmonary function in children in 2 Saskatchewan communities. AB - BACKGROUND: Asthma prevalence is known to vary among different geographical regions both nationally and internationally. However, there is limited understanding of the nature of differences within geographical regions. OBJECTIVE: To evaluate the prevalence of asthma in 2 prairie communities and differences in the patterns of respiratory symptoms between the communities. METHODS: A cross-sectional questionnaire survey was sent through schools in Estevan and Swift Current, Saskatchewan, to parents of 2,231 children in grades 1 to 6. Asthma prevalence was determined by questionnaire report of physician diagnosed asthma. Pulmonary function tests (PFTs) using spirometry were conducted in children in grades 1 to 4. To evaluate respiratory morbidity without the use of a diagnostic label, similar comparisons were made between communities for respiratory symptoms. RESULTS: The overall response rate to the survey questionnaire was 91.3%. The prevalence of ever asthma in Estevan was 21.4% (95% confidence interval [CI], 20.1%-22.7%) compared with 16.2% (95% CI, 15.1%-17.3%) in Swift Current. A higher proportion of girls in Estevan (19.7%; 95% CI, 17.9% 21.5%) compared with girls in Swift Current (12.5%; 95% CI, 11.1%-13.9%) reported a history of asthma. There was no difference found between towns for boys. These findings were supported by findings for respiratory symptoms, including wheeze and cough. For both boys and girls, the forced expiratory flow at 25% to 75% of forced vital capacity and the ratio of forced expiratory volume in 1 second to forced vital capacity were lower in Estevan compared with Swift Current. CONCLUSIONS: Differences in the distribution of childhood asthma can be found within regions. These results are strengthened by PFTs and cannot be fully explained by diagnostic biases. PMID- 14756466 TI - Adverse reactions of prophylactic intravenous immunoglobulin infusions in Iranian patients with primary immunodeficiency. AB - BACKGROUND: Although long-term intravenous immunoglobulin infusion is an effective treatment for children with antibody deficiencies, it can be complicated by systemic adverse reactions. OBJECTIVE: To evaluate the adverse reactions of intravenous immunoglobulin therapy in patients with primary immunodeficiency. METHODS: Seventy-one immunodeficient patients receiving intravenous immunoglobulin were evaluated during a 7-year period (1995-2002) at Children's Medical Center in Tehran, Iran. Immunological diagnoses were as follows: common variable immunodeficiency (31 patients), X-linked agammaglobulinemia (25 patients), IgG subclass deficiency (5 patients), hyper-IgM syndrome (2 patients), and ataxia-telangiectasia (8 patients). RESULTS: One hundred fifty-two cases (12.35%) of adverse reactions occurred following 1,231 infusions in 35 patients. The most frequent immediate adverse reactions were mild reactions (131 infusions), including chills, fever, flushing, muscle pains, nausea, headache, and anxiety. Moderate reactions, such as vomiting, chest pain, and wheezing, occurred in 19 infusions. Two patients experienced severe adverse reactions. The highest proportion (23.06%) of reaction to injection was in patients with common variable immunodeficiency. CONCLUSIONS: Intravenous immunoglobulin is a well tolerated medical agent for patients with antibody deficiency. However, to prevent occurrence of immediate adverse reactions during infusion in these patients, physicians should perform a detailed history and proper physical examination and check the titer of anti-IgA. PMID- 14756467 TI - The promotion of eosinophil degranulation and adhesion to conjunctival epithelial cells by IgE-activated conjunctival mast cells. AB - BACKGROUND: Allergen-mediated mast cell activation is a key feature of ocular allergic diseases. Evidence of eosinophil-derived mediators in tears and conjunctival biopsy specimens has been associated with chronic ocular allergic inflammation. OBJECTIVE: To examine the role of conjunctival mast cell mediators in eosinophil adhesion to conjunctival epithelial cells and eosinophil degranulation. METHODS: Conjunctival cells were obtained by enzymatic digestion of cadaveric conjunctival tissues. Eosinophils were obtained from peripheral blood samples using negative magnetic bead selection. The effect of IgE-activated mast cell supernates on eosinophil degranulation and adherence to epithelial cells was compared with supernates obtained from mast cells pretreated with a degranulation inhibitor (olopatadine). Eosinophil adhesion was measured by eosinophil peroxidase assay, and eosinophil degranulation was measured by eosinophil-derived neurotoxin radioimmunoassay. RESULTS: IgE-activated conjunctival mast cell supernates stimulated adhesion of eosinophils to epithelial cells (20.4% +/- 6.3% vs 3.1% +/- 1.0%; P = .048). Degranulation was not required for this process (no effect of olopatadine). IgE-activated mast cell supernates stimulated eosinophil-derived neurotoxin release (108.89 +/- 8.27 ng/10(6) cells vs 79.45 +/- 5.21 ng/10(6) cells for controls, P = .02), which was significantly inhibited by pretreatment of mast cells with a degranulation inhibitor (79.22 +/- 4.33 ng/10(6) cells vs 61.09 +/- 5.39 ng/10(6) cells for olopatadine pretreated and untreated, respectively, P = .02). CONCLUSIONS: Mediators released from conjunctival mast cells promote eosinophil adhesion to conjunctival epithelial cells and eosinophil degranulation. Degranulation inhibition studies suggest that different mast cell mediators are involved in regulation of these events. PMID- 14756468 TI - Comparison of the combinations of fexofenadine-pseudoephedrine and loratadine montelukast in the treatment of seasonal allergic rhinitis. AB - BACKGROUND: Antihistamine-decongestant combinations are used routinely for the treatment of seasonal allergic rhinitis. Recently, the combination of an antihistamine and a leukotriene receptor antagonist has been shown to be efficacious. OBJECTIVE: To compare the 2 combinations in the treatment of seasonal allergic rhinitis. METHODS: This was a randomized, double-blind, double dummy, parallel study in which patients with seasonal allergic rhinitis received either fexofenadine, 60 mg, and pseudoephedrine, 120 mg, twice daily, or loratadine, 10 mg, and montelukast, 10 mg, once daily, for 2 weeks. The Rhinoconjunctivitis Quality of Life Questionnaire (RQLQ) was completed at the beginning and end of the study. Patients recorded nasal symptoms and measured nasal peak inspiratory flow (NPIF) twice daily. Baseline measurements were obtained before initiation of treatment. RESULTS: Compared with baseline, both treatments resulted in statistically and clinically meaningful reductions of overall and individual RQLQ domain scores (P < .01) except for the sleep domain, for which only loratadine-montelukast led to significant improvement. There was a significant reduction in total symptoms (P < or = .05) compared with baseline on most treatment days in patients receiving both combinations. When the change from baseline was analyzed, there were no statistically significant differences in total symptoms between fexofenadine-pseudoephedrine and loratadine-montelukast (median, -28.5 vs -22.5; P = .33). There was a significant improvement in NPIF from baseline on all treatment days in both groups (P < .05), with no significant difference between treatments. CONCLUSIONS: Fexofenadine-pseudoephedrine and loratadine-montelukast have comparable efficacy in improving symptoms, RQLQ scores, and nasal obstruction in seasonal allergic rhinitis. The lack of improvement in sleep in the fexofenadine-pseudoephedrine group is probably related to insomnia, a known adverse effect of pseudoephedrine. PMID- 14756469 TI - Treatment of Churg-Strauss syndrome with high-dose intravenous immunoglobulin. AB - BACKGROUND: In some patients with Churg-Strauss syndrome (CSS), especially those with myocardial or neural involvement, conventional treatment with corticosteroids with or without cyclophosphamide is not effective. OBJECTIVE: To examine the effects of intravenous high-dose immunoglobulin (IVIG) in patients with CSS who showed poor responsiveness to conventional treatment. METHODS: We consecutively selected patients with CSS who showed any organ involvement despite corticosteroid treatment with or without cyclophosphamide. The diagnosis was based on the classification criteria of the American College of Rheumatology. IVIG therapy was performed with a dose of 400 mg/kg of immunoglobulin daily for 5 days. Neuropathy was evaluated with the manual muscle strength test and by the skin temperature of affected sites. Cardiac function was examined with ejection fraction by echocardiography and 2 imaging tests of myocardium (iodine 123 metaiodobenzylguanidine and thallium 201). RESULTS: The manual muscle strength test results were improved, and the skin temperature of both hands and legs was increased by IVIG therapy. In 5 patients with heart failure, the mean +/- SD ejection fraction of the left ventricle increased from 35.2% +/- 13.9% to 61.0% +/- 10.1% (P < .02). The uptake of iodine 123 metaiodobenzylguanidine of the myocardium increased, indicating that the myocardial viability was improved. The thallium 201 images revealed the presence of perfusion defects, which were improved by IVIG therapy. CONCLUSIONS: Patients with CSS who are resistant to corticosteroid treatment with or without cyclophosphamide may be treated effectively with IVIG therapy. PMID- 14756470 TI - Cellular immunodeficiency and autoimmunity in long-term mineral oil administration. AB - BACKGROUND: Subcutaneous mineral oil injection is an old-fashioned practice used mostly for cosmetic purposes. Infection, ulceration, subcutaneous nodules, and autoimmune activation are among the known adverse effects. Immunodeficiency has not been previously reported in association with mineral oil injection. OBJECTIVE: To report the case of a 43-year-old woman who performed long-term self administration of mineral oil and was found to have both cellular immunodeficiency and autoimmunity. METHODS: We performed an immunological evaluation. Throat, induced sputum, urine, and blood cultures were examined for microorganisms. Pelvic computed tomography, inguinal lymph node biopsy, bone marrow biopsy, and liver biopsy were also performed. RESULTS: Laboratory results revealed peripheral lymphopenia, very low absolute numbers of lymphocyte subpopulations, and a markedly impaired lymphocyte proliferative response to mitogens (phytohemagglutinin and concanavalin A) and recall antigens (mumps, Candida albicans, purified protein derivative, and tetanus toxoid). The cultures were negative for microorganisms. The pelvic computed tomogram demonstrated areas of diffuse oil-density signals throughout the subcutaneous tissue in the gluteal area and proximal lower extremities, as well as bilateral inguinal lymphadenopathy. A lymph node biopsy specimen showed lipid granulomas and necrotizing lymphadenitis. A bone marrow biopsy specimen demonstrated hypercellular marrow with normal trilineage hematopoesis. Increased serum transaminase levels, hypoalbuminemia, positive anti-extractable nuclear antigen and anti-Ro antibodies, and plasma cells in the liver suggested an autoimmune process. CONCLUSIONS: Mineral oil administration may be associated with both cellular immunodeficiency and autoimmunity. Patients who have received long-term administration of a foreign substance should undergo a comprehensive immunological evaluation. PMID- 14756471 TI - Allergy to fungal allergens in northern Italy. PMID- 14756472 TI - Prolonged survival in Churg-Strauss syndrome. PMID- 14756473 TI - Acute anaphylaxis after pine nut skin testing. PMID- 14756474 TI - Dry vs aqueous inhalers. PMID- 14756475 TI - The management of local reactions due to allergen immunotherapy. PMID- 14756476 TI - 'Plunging' during burr hole craniostomy: a persistent problem amongst neurosurgeons in Britain and Ireland. AB - The inadvertent 'plunging' of an instrument into the cranial cavity is a feared complication of drilling a burr hole and while anecdotes abound, little is known about the extent or the consequences of this problem. A survey by anonymous postal questionnaire of 304 neurosurgeons in Britain and Ireland was conducted to analyse the extent of this complication. Of respondents, 65.6% had experienced 'plunging', 22.3% having 'plunged' at least twice, indicating a high prevalence of this complication. The Cushing perforator was implicated by most. 'Plunging' carried a 12% risk of death or permanent neurological morbidity. The authors analyse the prevalence and significance of this preventable complication, and discuss various options available to minimize its occurrence. PMID- 14756477 TI - Spinal epidural abscesses: conservative treatment for selected subgroups of patients. AB - The aim of this study was to emphasize the importance of non-surgical treatment for subgroups of patients with spinal epidural abscesses (SEA). From 1988 to 2000, thirty cases of epidural spinal abscesses were retrospectively included in the study. The records and radiological studies were evaluated. Staphylococcus aureus was the most frequent microorganism causing SEA in 18 patients. In 20 patients SEA was secondary to interventional procedures. Predisposing factors were present in 15 cases. Fifty per cent was located in the lumbar region. A total of 22 patients received MRI, which always gave the diagnosis of SEA. Surgical treatment was performed in 20 patients. Conservative treatment with antibiotics was used in 10 patients. Eight patients did not have neurological deficits. One patient was critically ill and another patient was paralysed with an epidural lesion extending over six spinal segments. In all cases, a microorganism was known at the time of diagnosis of ESA. The eight patients without deficits recovered completely following treatment with antibiotics. C reactive protein was the most reliable inflammatory marker to monitor the effect of the treatment. MRI enables diagnosis of ESA before deficits occur. These can safely be treated with antibiotics if the causative microorganism is known, and the neurological status and laboratory values are monitored. Decompressive surgery is restricted to cases with progressive deficits, when the deficits have lasted for less than 36 h and when the microorganism is not known. PMID- 14756478 TI - Spinal dural arteriovenous fistulas: the use of intraoperative microvascular Doppler monitoring. AB - Spinal dural arteriovenous fistulas are characterized by an abnormal communication between the dural branch of the radiculomedullary artery and an intradural medullary vein. Although the optimal treatment strategy is still debated, a complete interruption of the flow in the fistulas should be obtained. The authors report four cases operated on with intraoperative microvascular Doppler monitoring assistance. In all cases, microDoppler confirmed the location of the fistula and revealed an arterial spectrum on the redundant dorsal medullary veins. After the clipping of the feeder of the arteriovenous shunt, the intraoperative monitoring documented a complete disappearance of the arterial spectrum and the reappearance of the venous pattern. The ultrasonographic changes suggested the complete interruption of the fistulas. Postoperative angiography showed no residual abnormality in all patients. Doppler monitoring during surgery confirms satisfactory interruption of the arterial feeder and may prove useful where initial identification of the feeding vessel is difficult. PMID- 14756479 TI - Prognostic importance of transferrin receptor expression and correlation with K1 67 labelling indices in intracranial meningiomas. AB - Meningiomas are variously benign, atypical or anaplastic neoplasms and can be treated by surgical removal. However, recurrence can be seen even after complete surgical resection in benign meningiomas and some are histologically aggressive. As predictors of recurrence or malignant proliferation some immunohistochemical markers have been used. In this study, we postoperatively identified TfR (transferrin receptor) staining and Ki-67 proliferative index in patients with intracranial meningiomas and evaluated the correlation between these parameters and the recurrence or malignant proliferation. Immunohistochemical techniques (streptavidin-biotin complex) were used to assess the TfR expression and Ki-67 labelling index in 50 surgically removed intracranial meningiomas. Significantly high TfR expression was observed in all types of meningiomas, eight of which recurred. Four cases died because of primary intracranial pathology and one died from uncontrollable epileptic seizures. Ki-67 levels were high in the cases which showed recurrence and showed atypical features. Based on our observations and the results presented above, meningioma patients with TfR score of 3 or higher and high Ki-67 labelling index must be carefully followed up for recurrence, as well as for malignant transformation. Thus, we suggest that TfR and Ki-67 immunostains should be applied routinely in patients with meningiomas. PMID- 14756480 TI - MRI study of the natural history and risk factors for pseudomeningocoele formation following postfossa surgery in children. AB - Surgical approaches to the posterior fossa may be complicated by pseudomeningocoele formation. We report on its natural history and risk factors for its formation, as seen on serial MRI postoperatively in children with posterior fossa tumours. In a retrospective study of 84 children undergoing surgery for posterior fossa tumours, 13 (16%) developed clinically apparent pseudomeningocoeles. On postoperative MRI, pseudomeningocoeles were apparent in 34 (41%) patients at 1-5 days, but in only four patients at 10-15 months postsurgery. There was a progressive decrease in the mean depth of pseudomeningocoele measured from the MRI scans postoperatively. Patients with pseudomeningocoeles were more likely to have a postoperative CSF leak from the wound (39 v. 13%), lumbar punctures or lumbar drains (54 v. 25%), wound re closures (23 v. 1%) and prolonged hospital stay (19.9 v. 14.5 days). On multivariate analysis, patients with pseudomeningocoeles were also more likely to have undergone a suboccipital craniectomy than those without pseudomeningocoeles (69 v. 38%). Postoperative pseudomeningocoele formation following posterior fossa surgery is more apparent radiologically than clinically, but there is clinical and radiological evidence that pseudomeningocoeles gradually resolve over the postoperative period. The risk of pseudomeningocoele formation is increased by performing a suboccipital craniectomy and there is an association with increased CSF leaks, needing re-closure of the wounds. PMID- 14756481 TI - Rapid clinical deterioration in a patient with multi-focal glioma despite corticosteroid therapy: a quantitative MRI study. AB - A patient with high-grade multi-focal glioma deteriorated rapidly despite high dose corticosteroid therapy (dexamethasone: 16 mg/day). MRI was used to measure diffusion tensor parameters and longitudinal relaxation time (T1) values of peritumoural oedematous brain before and after commencing steroid treatment. Forty-eight hours after steroid treatment there was no evidence of brain oedema reduction. Specifically, regions of oedematous brain showed a significant increase in mean diffusivity () with a significant decrease in diffusion anisotropy (p < 0.05), but without any change in T1 values. These quantitative MRI data were mirrored by the rapid deterioration seen when assessing the patient clinically. This case shows that quantitative MRI can not only measure steroid treatment response but also failure in patients with malignant gliomas. PMID- 14756482 TI - Treatment of orbital schwannomas and neurofibromas. AB - We present an overview of the treatment and clinical outcome of five orbital peripheral nerve tumours, carried out in our centre from 1999 to 2003. The surgical approach was determined by the location and extension of the lesion. Supraorbital orbitotomy was performed in two superiorly located lesions, a transconjunctival approach in one medial, basal, extraconal lesion. A pterional extradural approach was used in two cases with involvement of the apex, superior orbital fissure and cavernous sinus. Three patients were diagnosed as having schwannoma, one as neurofibroma, and one as cystic mixed neurofibroma and schwannoma. One patient suffered from multiple schwannomas [bilateral acoustic schwannomas, cervical schwannomas (NF2)]. One patient showed bilateral orbital neurofibromas, plexiform cutaneous neurofibroma (NF1) and glaucoma due to a coexisting Marfan's syndrome. Local recurrences were not seen after complete resection in all patients. Surgery is the therapeutic goal. PMID- 14756483 TI - Syringe tip accuracy for application of Victor Horsley's bone wax. AB - An inexpensive way of improving the handling properties of white bone wax is described. PMID- 14756484 TI - Carbon dioxide insufflation for chronic subdural haematoma: a simple addition to burr-hole irrigation and closed-system drainage. AB - Burr-hole irrigation with closed-system drainage is a common surgical method used for chronic subdural haematoma. However, the subdural space with air that entered during surgery sometimes remains for a prolonged period after surgery and may hamper uncomplicated healing of the subdural space. We combined a simple procedure, insufflation of carbon dioxide (CO2) into the subdural space through a drainage catheter, with conventional burr-hole irrigation and closed-system drainage. By this additional procedure, both the subdural space and the gas within the space decreased rapidly, and the subdural drain could be removed within 24 h. By promoting obliteration of the subdural space, this simple combined technique may contribute to early recovery and discharge of patients, and to a reduction in the recurrence rate of the disease. PMID- 14756485 TI - A new tumour forceps for use during removal of pineal region tumours. AB - A new forceps for grasping and cutting tumour in a narrow surgical field is described. The working length is 12 cm and the grasping element at the tip is 1 mm in diameter. To avoid damage to surrounding structures caused by pulling out the tumour, the grasping portion consists of two hollow cylinders with sharp edges that divides, rather than tears tissue. PMID- 14756486 TI - Reconstruction of the sella floor using vascularized pedicled mucosal flap. AB - The efficacy and uses of a vascularized pedicled sphenoid sinus mucosal flap in reconstruction of the sella floor after trans-sphenoidal surgery for sellar lesions is studied. The method was successfully performed in 31 cases over a period of 7 years. The ease of harvesting, high vascularity and local availability of the flap are the principal advantages. PMID- 14756487 TI - Vertebral cryptococcosis simulating tuberculosis. AB - Infection with the fungus Cryptococcus neoformans is seen predominantly in two forms: (a) pulmonary and (b) cerebromeningeal. Skeletal cryptococcosis is uncommon. There have been only occasional case reports of thoracic vertebral cryptococcosis presenting as cord compression. A young female had cervical lymphadenopathy diagnosed as tuberculosis by fine needle aspiration cytology (FNAC) and was on antitubercular therapy (ATT) for 5 months. She developed rapidly progressive paraparesis and imaging demonstrated a destructive vertebral body lesion involving T2-3 with a paraspinal abscess producing cord compression. A costotransversectomy with excision of the diseased bone and bone grafting was done. Histopathological examination revealed cryptococcosis. The patient was put on antifungal medication, but expired 2 weeks after surgery. Radiological, magnetic resonance imaging and surgical finding of vertebral cryptococcosis can mimic tuberculosis. The definite diagnosis of cryptococcosis depends upon microscopic identification of the organism. A high index of suspicion leading to early surgical confirmation and institution of anti fungal therapy is necessary to reduce the mortality and morbidity. PMID- 14756488 TI - Haemangiopericytoma presenting as acute subdural haematoma. AB - Haemangiopericytomas are vascular neoplasms that arise from capillary pericytes. They rarely present with haemorrhage. We report a patient whose meningeal tumour was revealed by acute subdural and intratumoural haemorrhage. PMID- 14756489 TI - Haemangioma calcificans. AB - Haemangioma calcificans is a relatively rare intracranial tumour, which is characterized by the presence of a calcified nodule in or near the brain. The patient is most often an adult of either sex, who develops seizures. The lesion can be large enough to produce raised intracranial pressure. The location of tumour is subcortical and often in the temporal lobe. It is usually solitary and benign in nature. Microscopically, there are thin-walled blood vessels separated by dense fibrous bands containing deposits of haemosiderin and calcium. On electron microscopy, tubular structures limited by a wall of electrodense material and irregular calcium deposits within them are prominent features. Four cases of haemangioma calcificans are described here with review of literature. PMID- 14756490 TI - Endovascular stenting of the transverse sinus in a patient presenting with benign intracranial hypertension. AB - The authors present a 37-year-old lady with symptoms and signs suggestive of benign intracranial hypertension (BIH). Routine CT and MRI scans were normal. Further investigations were performed with magnetic resonance venography (MRV) and cerebral venography. These revealed obstruction of the right transverse sinus with high pressure (40 mmHg) proximal to the obstruction and low pressure (15 mmHg) distally. She was treated by transvenous stent deployment with resolution of her symptoms and the bilateral papilloedema. Evaluation of the cerebral venous system with MRV and or with formal cerebral venography should be included in routine investigations of patients with suspected BIH. PMID- 14756491 TI - Melioidosis presenting as spinal epidural abscess. AB - Central nervous system melioidosis is an unusual infection in humans. This article reports a case of melioidosis presenting as an acute spinal epidural abscess. A discussion of this case and its management together with a brief review of melioidosis of the central nervous system is presented. PMID- 14756492 TI - Overwhelming cranial and spinal subdural empyema secondary infected sacral decubitus ulcers. PMID- 14756493 TI - The importance of angiography before exploring a Sylvian fissure haematoma: a case report showing the quality of CT angiography. PMID- 14756495 TI - Massive cerebral cyst complicating hypertensive stroke. PMID- 14756494 TI - Unusually widened perivascular spaces in the cerebral hemisphere: the importance of MRI for diagnosis of a benign clinical entity. PMID- 14756496 TI - The investigation of acute severe headache suggestive of probable subarachnoid haemorrhage: a hospital-based study. AB - We reviewed the diagnostic approach to patients presenting with headache suggestive of subarachnoid haemorrhage (SAH) and normal cranial imaging, both locally and using a questionnaire, in other units throughout the United Kingdom. We emphasize the correct timing of cranial imaging and lumbar puncture (LP), the importance of proceeding to LP and utilizing spectrophotometry in patients with normal cranial imaging, and the appropriate use of angiography. PMID- 14756497 TI - Attitudes of people with disabilities toward physician-assisted suicide legislation: broadening the dialogue. AB - This article presents the methods, findings, and implications of a participatory action research project that attempted to shed additional light on the debate over death with dignity (DWD) or physician-assisted suicide (PAS) legislation. In depth, qualitative interviews with forty-five physically disabled residents of the San Francisco Bay Area, conducted by others with disabilities, revealed a wide breadth of opinions about and attitudes toward such legislation. For close to half of the participants, the desire for autonomy in making end-of-life decisions was a primary concern, yet fear that PAS legislation could violate this autonomy in various ways was a deep concern as well. Also reported were widespread accounts of disability-based discrimination and frequent expressions of fear about openly discussing positions that diverge from the official, publicly held opinions of disability leaders who oppose such legislation. The findings support those of a recent Harris poll demonstrating considerable diversity of opinion about PAS legislation among people with disabilities. The findings further suggest the need for additional research on the apparent disjunction between the diversity of attitudes held by those interviewed and the more unified position taken by many disability activists. Use of the study findings to promote greater dialogue within the community and to better position people with disabilities to take their place at the policy table also is discussed. In addition, the findings are seen as reinforcing the need for the public health community to become more engaged in this central ethical debate. PMID- 14756498 TI - Commodifying life? A pilot study of opinions regarding financial incentives for organ donation. AB - In recent years many policy proposals have been put forward to create financial incentives to encourage families to allow the harvesting of organs from their deceased relatives. While research has focused on whether these policies would actually increase the supply of organs, no research has focused on testing the ethical concerns about such policies. This article presents the findings of a pilot study conducted to determine whether people think that families should end life support of a family member in order to harvest organs if various incentive policies are in place. While the findings do not suggest a direct effect of these inducements, they do indicate that the amount of money received from organ donation is a consideration in making the decision whether to end life support. The implications of this finding for ethical debates and health policy are reviewed. PMID- 14756499 TI - Racial meanings and scientific methods: changing policies for NIH-sponsored publications reporting human variation. AB - Conventional wisdom holds that race is socially constructed and not based on genetic differences. Cutting-edge genetic research threatens this view and hence also endangers the pursuit of racial equality and useful public health research. The most recent incarnation of racial genetics is not due to scientific discoveries about population differences per se, but follows from how the United States and other governments have organized racial categories. This article explains tensions in U.S. government guidelines and publications on the study of human genetic diversity, points out the absence of any compelling public health benefits that might justify this research, introduces conceptual tools for addressing the complicated heuristic and policy problems posed by medical population genetics, and offers two policy proposals to remedy the current problems. PMID- 14756500 TI - The many faces of access: reasons for medically nonurgent emergency department visits. AB - Investigating why people use the hospital emergency department (ED) for visits considered medically nonurgent can enhance our understanding of people's expectations of health care services, of their conceptions of prudent lay judgment, and of difficulties in negotiating the logistics of primary care services. This study identified reasons for such ED use from users' perspectives in both pediatric and adult visits. Respondents were asked to explain what brought them to the ED and to define an emergency. The study was conducted in two northeastern U.S. hospital EDs. The analysis drew on a convenience sample of 408 (331 pediatric, 77 adult users) face-to-face interviews that employed both open- and closed-ended questions. Findings indicate most patients had medical insurance and a regular place of care and most arrived by car or taxi. Twelve main themes emerged under three main categories: conceptions of needs, appropriateness, and preference for the ED. The findings indicate that various reasons for ED use may be construed as access issues. These include beliefs regarding limited availability of after-hour consultation services and of timely appointments at one's primary care site. Drawing on the findings, a typology that distinguishes between groups of users according to their preference for the ED, a level of congruence between their own reason and their definition of an emergency was developed. The typology suggests that people's concerns that influence their decision to come to the ED cannot be solved simply by expanding primary care services or by educational interventions. Its application yields recommendations for services and interventions. PMID- 14756501 TI - RCVS gathers new ideas on VN training. PMID- 14756502 TI - Endoscopic renal evaluation and biopsy of Chelonia. AB - Sixty-nine tortoises, turtles and terrapins representing 28 species of the order Chelonia, class Reptilia were evaluated by endoscopy for renal disease. Under general anaesthesia, coelomic and/or extracoelomic endoscopic evaluations and biopsies of the kidney(s) were undertaken. Endoscopic approaches required a 2 to 4 mm skin incision in the prefemoral fossa, and minimal blunt dissection through the subcutaneous tissues. For the coelomic approach the coelomic aponeurosis of the transverse and oblique abdominal muscles was penetrated so that the cranioventral kidney(s) could be examined and biopsied. The extracoelomic approach required the endoscope to be advanced in a caudodorsal direction, between the coelomic aponeurosis and the broad iliacus muscle, so that the dorsolateral kidney(s) could be examined and biopsied. Both techniques were safe and effective for obtaining renal biopsies for histological examination and microbiological culture. Several renal pathologies were identified including glomerulonephrosis, tubulonephrosis, interstitial nephritis, uric acid accumulation, tubulonephritis, glomerulonephritis, renal oedema, glomerulosclerosis, nephrosclerosis, soft tissue mineralisation, and pyelonephritis. Several infectious conditions were identified, including a predominance of Gram-negative bacterial infections, two cases of hexamitiosis, and one case of mycobacteriosis. PMID- 14756503 TI - Studies on the carriage and transmission of porcine reproductive and respiratory syndrome virus by individual houseflies (Musca domestica). AB - The objectives of the study were to determine the site of porcine reproductive and respiratory syndrome virus (PRRSV) in individual houseflies, to assess whether an individual housefly could transmit PRRSV to a susceptible pig, and to compare the ability of PCR, virus isolation and a pig bioassay to detect PRRSV in houseflies. In the first experiment 26 houseflies were fed on a pig infected experimentally with PRRSV; 13 were processed as a whole fly homogenate, while an exterior surface wash and a gut homogenate were collected from the other 13. Infectious PRRSV was recovered from nine of the whole fly homogenates, 12 of the gut homogenates and one of the exterior surface washes. In the second experiment, two of 10 individual houseflies, which had fed on an infected pig, transmitted PRRSV to a susceptible pig in a controlled manual transmission protocol. In the third experiment, single flies or pools of 30 flies were immersed in different concentrations of a PRRSV inoculum, then tested by PCR, virus isolation and bioassay. The virus was detected at a concentration of 10(1) TCID50/ml by PCR, 10(2) TCID50/ml by the bioassay and 10(3) TCID50/ml by virus isolation. PMID- 14756504 TI - Estimation of the liveweight of working mules in Morocco from their body measurements. AB - During March 2001, 117 working mules, randomly chosen from four markets in Morocco, were weighed and a series of body measurements recorded; their age and body condition score were also recorded. Linear regression techniques were used to derive a 'best fit' equation for predicting liveweight from the other variables. For the working mules weighing between 131 and 391 kg the best prediction equation using two variables was: liveweight (kg) = -33 + 2.8 x heart girth (cm)+1.36 x length (cm). Other prediction equations available for estimating the liveweight of equidae were tested on the data, but in all cases they significantly overestimated the weights of the Moroccan mules. PMID- 14756505 TI - Emergence of new subclones of multiresistant Salmonella typhimurium DT104 possibly associated with poultry meat. PMID- 14756506 TI - Pilot study of intraregional deionised water adjunct therapy for mast cell tumours in dogs. PMID- 14756507 TI - Emergence of an apparently neurotropic maedi-visna virus infection in Britain. PMID- 14756508 TI - Guidance for Named Veterinary Surgeons. PMID- 14756509 TI - Veterinary pathology. PMID- 14756510 TI - Future of farm animal practice. PMID- 14756511 TI - Dairy cow diets and milk production. PMID- 14756512 TI - Identification and microchips. PMID- 14756513 TI - Anthelmintic resistance and use of anthelmintics in horses. PMID- 14756514 TI - Training for veterinarians in organic systems. PMID- 14756515 TI - Control of bovine TB. PMID- 14756516 TI - Care of military working dogs. PMID- 14756517 TI - Melanosome transfer to and translocation in the keratinocyte. AB - Complexion coloration in humans is primarily regulated by the amount and type of melanin synthesized by the epidermal melanocyte. However, additional and equally contributing factors consist of (1) efficient transfer of melanin from the melanocytes to the neighboring keratinocytes and (2) distribution and degradation of the transferred melanosomes by the recipient keratinocytes. Once synthesized in the cell body of the epidermal melanocyte, pigmented melanosomes are translocated down the dendrites and captured at the dendritic tips via various cytoskeletal elements. Molecules recently identified that participate in this process consist of Rab27a, myosin-Va and melanophilin. Eventually, these peripherally localized melanosomes are transferred to keratinocytes by a presently undefined mechanism. The protease-activated receptor-2 (PAR-2) and unidentified surface lectins and glycoproteins facilitate this transfer process. Once incorporated into the keratinocytes, melanosomes are distributed individually or as clusters, aggregated towards the apical pole of the nucleus, and degraded as the keratinocytes undergo terminal differentiation and desquamation. Ultraviolet irradiation (UVR) can modulate the process of melanosome transfer from the melanocytes to the keratinocytes. UVR can upregulate expression of PAR-2 and lectin-binding receptors and increase phagocytic activity of cultured keratinocytes. Therefore, many cellular and molecular events that occur after melanogenesis contribute to skin color. PMID- 14756518 TI - Effect of sunlight exposure and aging on skin surface lipids and urate. AB - Free fatty acids (FFA), squalene, squalene hydroperoxide, and uric acid in the methanol extracts from human skin surface were measured. Levels of FFA and squalene were significantly lower in the older (83.7 +/- 9.4 years) than in the younger (22.2 +/- 3.9 years) group. FFA are mostly saturated, and linoleic acid is an exclusive polyunsaturated fatty acid. The composition of linoleic acid decreased in the older group by 40%, suggesting age-dependent loss of oxidatively vulnerable polyunsaturated fatty acid. Even monounsaturated acids such as palmitoleic and oleic acids decreased significantly in the older group. This could be interesting because 2-nonenal is the oxidation product of palmitoleic acid and has been identified as the major aged body odor component. Sunlight exposure for 1.5 h did not change levels of FFA and squalene, or FFA composition. However, squalene hydroperoxide increased by 60-fold, as reported previously, suggesting that hydroperoxide is produced by singlet oxygen. Uric acid increased by two-fold, which may be the adaptive response against photo-oxidative stress because uric acid is a good scavenger of singlet oxygen and oxygen radicals. PMID- 14756519 TI - Photoaging and oxidative stress. AB - Photoaging is significantly different from chronological aging in both clinical and histological appearance. It has been suggested that oxidative stress, generated by ultraviolet radiation (UVR), leads to photoaging over a long period. The presence of 8-OHdG, and oxidatively modified proteins such as 4-hydroxy-2 nonenal-modified protein, 3-L-nitro-tyrosine and N(-epsilon) (carboxymethyl)lysine in UV-exposed skin specimens, supports this theory. The pathophysiology of photoaging of the skin caused by chronic inflammation after UVR is reviewed and discussed, with a focus on oxidative stress. PMID- 14756520 TI - Involvement of changes in stratum corneum keratin in wrinkle formation by chronic ultraviolet irradiation in hairless mice. AB - Chronic exposure to ultraviolet (UV) radiation alters components of the skin. We previously reported that chronic low-dose UV irradiation induced wrinkle formation but did not significantly alter dermal components of hairless mice. In the present study, we examined whether the epidermal changes by UV irradiation could contribute to wrinkle formation. Hairless mice were irradiated with one third the minimum erythema dose (MED) of UVB (20 mJ/cm2) and UVA (14 J/cm2) for 10 weeks, and followed up for another 24weeks without irradiation. Fine wrinkles were detected following irradiation, which existed even 24 weeks after cessation of irradiation. An increase in the viscoelastic proportion of the total distension (Uv/Ue) was recognized in irradiated mice, which may be related to wrinkle formation. In the epidermis, an increase in the amount of transepidermal water loss and a decrease in the water content of the stratum corneum were seen after 10 weeks of UV irradiation, neither of which recovered during the 24-week observation period. In addition, there was a significant increase in the amount of stratum corneum keratin after 10 weeks of irradiation, which persisted during the 24-week follow-up. The results of our study suggest that chronic low-dose UV irradiation primarily alters the epidermal rather than the dermal components of the skin. In addition, our results indicate that the increased keratin content of the stratum corneum may be involved in the alteration of the physical properties of the skin. This process could be one of the early events of wrinkle formation. PMID- 14756521 TI - Differential expression of heparin-binding EGF-like growth factor (HB-EGF) mRNA in normal human keratinocytes induced by a variety of natural and synthetic retinoids. AB - It was recently revealed that epidermal growth following topical treatment with all-trans retinoic acid (atRA) was at least partly induced by heparin-binding epidermal growth factor-like growth factor (HB-EGF) released from suprabasal keratinocytes. Since proliferation of keratinocytes appears to be one of the critical roles of atRA in depigmentation treatment and promotion of wound healing, HB-EGF is considered suitable for assessing the therapeutic value of topical retinoids. In this study, HB-EGF mRNA expression in normal human keratinocytes after atRA treatment was examined, and the effects of a variety of natural and synthetic retinoids were compared. The results of reverse transcription polymerase chain reaction (RT-PCR) suggested that induction of differentiation increased HB-EGF mRNA expression in cultured keratinocytes. Real time PCR analyses revealed that HB-EGF mRNA expression was elevated dose dependently with atRA, peaking at 12 h. This elevation was more prominent in confluent keratinocytes than in subconfluent cells, suggesting that differentiated keratinocytes are more subject to stimulation of HB-EGF expression by atRA than proliferating keratinocytes. HB-EGF mRNA was upregulated in differentiation-induced keratinocytes by all retinoids used in this study at 1 micromol/l, and marked upregulation was seen when treated with three isotypes of retinoic acid (atRA, and 9-cis and 13-cis retinoic acid). RARalpha-selective agonists (Am80, Am580, ER-38925, and TAC-101) and a panagonist of RARs (Re80) caused relatively low elevation of HB-EGF transcripts, as did all-trans retinol (Rol) and all-trans retinal (Ral). Although another panagonist (Ch55) showed the highest elevation of HB-EGF mRNA, it was relatively cytotoxic at the concentration employed. Ral and Rol were found to upregulate HB-EGF when used at 100 micromol/l to 1 mmol/l, to a similar extent of atRA at 1-10 micromol/l. The capacity of retinoids to upregulate HB-EGF may be an important index for investigation and development of an ideal synthetic retinoid, which has maximum benefits and minimum side-effects PMID- 14756522 TI - Tretinoin reverses upregulation of matrix metalloproteinase-13 in human keloid derived fibroblasts. AB - Keloids are skin abnormalities that are characterized by excessive deposition of collagen bundles in the dermis. Patients with keloids complain not only about their cosmetic appearance, but also about continuous itching and/or tenderness associated with chronic inflammation. Degradation of extracellular matrix (ECM) may be upregulated, associated with the expansion of keloids into circumferential skin, and high metabolic activity of keloid tissues may be due to increased matrix metalloproteinase (MMP) activity. Based on these hypotheses, we examined differences in expression of MMP-1, MMP-8, and MMP-13 between keloid-derived and normal dermal fibroblasts. Since retinoids are potent inhibitors of MMPs in the treatment of photoaged skin and cancers, we also examined whether or not tretinoin affects MMP expression of keloid-derived fibroblasts. The results of real-time polymerase chain reaction and ELISA demonstrated significant upregulation of MMP-13 and significant downregulation of MMP-1 and MMP-8 in keloid-derived fibroblasts, at both mRNA and protein levels. MMP-1 mRNA expression in the control group was significantly upregulated after the addition of tretinoin, whereas no significant change was observed in the keloid group. MMP 8 mRNA expression in the control group was significantly upregulated by tretinoin, with the peak at 12 h, while no significant change was observed in the keloid-derived fibroblasts. In contrast, the remarkably elevated MMP-13 mRNA expression in the keloid group was significantly suppressed, with the peak suppression at 12 h after addition of tretinoin, while MMP-13 mRNA expression in the control group was not significantly changed. The decrease in MMP-1 and MMP-8 may contribute to accumulation of type I and type III collagen in keloid tissues, and this mechanism may be modulated by molecular interaction with MMP-13. Tretinoin appeared to reverse the abnormal expression profile of MMPs in keloid derived fibroblasts, such as markedly elevated expression of MMP-13, partly through inactivation of AP-1 pathway. The present results suggest that tretinoin may be clinically useful to improve the chronic inflammation seen in keloids and prevent expansion of keloid tissues into circumferential normal skin. PMID- 14756523 TI - The inhibitory effect of glycolic acid and lactic acid on melanin synthesis in melanoma cells. AB - Alpha-hydroxy acids (AHAs) such as glycolic acid (GA) and lactic acid (LA) have been reported to be effective in treating pigmentary lesions such as melasma, solar lentigines, and postinflammatory hyperpigmentation. The mechanism of this effect might be due to epidermal remodeling and accelerated desquamation, which would result in quick pigment dispersion. However, the direct effect of AHAs on melanin synthesis has not yet been well studied. To elucidate such a direct effect of AHAs on melanogenesis, we performed melanin assays, growth curve determinations, Northern and Western blotting for melanogenic proteins [tyrosinase, tyrosinase related protein (TRP)-1 and TRP-2], and tyrosinase and, 4 dihydroxyphenylalaninechrome tautomerase enzyme activity assays using mouse B16 and human melanoma cells. GA or LA (at doses of 300 or 500 microg/ml) inhibited melanin formation in similar dose-dependent manner, without affecting cell growth. Although the mRNA and protein expression or molecular size of tyrosinase, TRP-1 and TRP-2 were not affected, tyrosinase activity was inhibited. To see whether GA and/or LA directly inhibit tyrosinase catalytic function, the effect of GA and LA on human tyrosinase purified from the melanosome-rich large granule fraction of human melanoma cells was performed. GA or LA were shown to inhibit tyrosinase enzyme activity directly, but this effect was not due to the acidity of GA or LA, because adjusting the pH to 5.6 (the pH of GA and LA at concentrations of 2500 microg/ml), did not affect tyrosinase activity. Taken together, these results show that GA and LA suppress melanin formation by directly inhibiting tyrosinase activity, an effect independent of their acidic nature. GA and LA might work on pigmentary lesions not only by accelerating the turnover of the epidermis but also by directly inhibiting melanin formation in melanocytes. PMID- 14756524 TI - Tobacco smoke extract induces age-related changes due to modulation of TGF-beta. AB - We have recently shown that tobacco smoking, like ultraviolet A radiation, is an important factor contributing to premature skin aging. We found that tobacco smoke extract decreased type I and III procollagen, increased tropoelastin mRNA, and induced abnormal accumulation of proteoglycans and matrix metalloproteinase (MMP)-1 and MMP-3 in cultured skin fibroblasts. This indicated that common molecular features might underlie the premature aging of the skin induced by tobacco smoke extract, including abnormal regulation of extracellular matrix deposition through elevated MMPs, reduced collagen production, abnormal tropoelastin accumulation, and altered proteoglycans. With the exception that reactive oxygen species were mediated in the aging process, transforming growth factor (TGF)-beta1 was found to play a crucial role in the age-related alterations induced by tobacco smoke extract. Here we report that tobacco smoke extract blocks cellular responsiveness to TGF-beta1 through the induction of a non-functional latent form of TGF-beta1, and downregulation of the TGF-beta1 receptor. This paper shows the evidence for the role of tobacco smoking in skin aging and describes how modulation of TGF-beta1 levels might retard premature skin aging. PMID- 14756525 TI - Biological effects of glycolic acid on dermal matrix metabolism mediated by dermal fibroblasts and epidermal keratinocytes. AB - Glycolic acid (GA), one of the alpha-hydroxy acids, is widely used as an agent for chemical peeling. Although there are several reports about the clinical effects of GA in the literature, its biological mechanism remains mostly unclear, and there are only a few reports about its effects on skin rejuvenation mediated by keratinocytes. The aim of this study was to investigate the effect of GA on the dermal matrix metabolism of keratinocytes and fibroblasts using in vitro and ex vivo systems. Our study shows that GA not only directly accelerates collagen synthesis by fibroblasts, but it also modulates matrix degradation and collagen synthesis through keratinocyte-released cytokines. We confirm that IL-1alpha is one of the primary mediators for matrix degradation released from keratinocytes after GA treatment. These results suggest that GA contributes to the recovery of photodamaged skin through various actions, depending on the skin cell type. PMID- 14756526 TI - The use of reconstructed human skin to evaluate UV-induced modifications and sunscreen efficacy. AB - Biological and clinical effects of sun exposures are characterized by short-term reactions, i.e. sunburn reaction and suntan, as well as long-term consequences corresponding to photoaging and photocancers. We have developed several human in vitro three-dimensional models in order to assess both the photodamage and the photoprotection afforded by sunscreens. Using a full thickness reconstructed skin comprising a differentiated epidermis and a living dermal equivalent, UVB- and UVA-induced biological markers could be found at both the keratinocyte and the fibroblast level. Typical markers of the sunburn reaction could be reproduced in that model as well as dermal damages related to the photoaging process. Another model of reconstructed epidermis, comprising keratinocytes but also melanocytes and Langerhans cells, has been developed. The study of the UV-induced pigmentation as possible using the pigmented reconstructed epidermis and allowing to reproduce the epidermal melanin unit. The assessment of cellular parameters related to UV-induced immunosuppression could be performed using the reconstructed epidermis containing Langerhans cells. Exposure to solar-simulated radiation provokes morphological alterations and the reduction in numbers of Langerhans cells within the exposed epidermis. Using all these models, the efficiency of sunscreens could be envisaged after topical application. The results showed that appropriate sunscreens could efficiently prevent the damage described above. PMID- 14756527 TI - Use of group-specific and RAPD-PCR analyses for rapid differentiation of Lactobacillus strains from probiotic yogurts. AB - The increasing interest in probiotic lactobacilli implicates the requirement of techniques that allow a rapid and reliable identification of these organisms. In this study, group-specific PCR and RAPD-PCR analyses were used to identify strains of the Lactobacillus casei and Lactobacillus acidophilus groups most commonly used in probiotic yogurts. Group-specific PCR with primers for the L. casei and L. acidophilus groups, as well as L. gasseri/johnsonii, could differentiate between 20 Lactobacillus strains isolated from probiotic yogurts and assign these into the corresponding groups. For identification of these strains to species or strain level, RAPD profiles of the 20 Lactobacillus strains were compared with 11 reference strains of the L. acidophilus and L. casei group. All except one strain could be attributed unambigously to the species L. acidophilus, L. johnsonii, L. crispatus, L. casei, and L. paracasei. DNA reassociation analysis confirmed the classification resulting from the RAPD-PCR. PMID- 14756528 TI - Cloning and expression of pyrroloquinoline quinone (PQQ) genes from a phosphate solubilizing bacterium Enterobacter intermedium. AB - A grass rhizosphere bacterium, Enterobacter intermedium (60-2G), has a strong ability to solubilize insoluble phosphate. Certain phosphate-solubilizing bacteria secrete gluconic acid for this process. The gluconic acid is produced by direct extracellular oxidation of glucose by a glucose dehydrogenase equipped with pyrroloquinoline quinone (PQQ) as a cofactor. A pqq gene cluster producing PQQ was detected in E. intermedium and this sequence conferred phosphate solubilizing activity to Escherichia coli DH5alpha. The 6,783-bp pqq sequence had six open reading frames (pqqA, B, C, D, E, and F) and showed 50-95% homology to pqq genes of other bacteria. E. coli DH5alpha expressing the E. intermedium pqq genes solubilized phosphate from hydroxyapatite after a pH drop to pH 4.0, which paralleled in time the secretion of gluconic acid. We speculate that production of PQQ in E. coli DH5alpha expressing the pqq cluster activates an endogenous glucose dehydrogenase to permit gluconic acid secretion that solubilizes the insoluble phosphate. PMID- 14756530 TI - Differentiation of Nostoc flagelliforme and its neighboring species using fatty acid profiling as a chemotaxonomic tool. AB - In this study, fatty-acid content and patterns were analyzed in order to distinguish Nostoc flagelliforme, an edible terrestrial cyanobacterium, from other Nostoc species and representatives typical of its close neighbors (genera Anabaena, Microcystis, and Synechococcus). According to the Kenyon-Murata classification system, all the Nostoc species were assigned to Group II due to the presence of C18:2n3 and C18:3n3, and the absence of C18:3n6. Hierarchical cluster analysis was also employed to separate N. flagelliforme and other Nostoc species or strains. A dendrogram calculation of all fatty-acid components manifested phenetic characteristics, showing that the degree of relatedness of two strains of N. flagelliforme aggregated them within a small subgroup. Another dendrogram, calculated from seven comprehensive parameters (including ratios of different fatty-acid categories, degree of fatty-acid unsaturation, etc.), also clearly delimited the minute difference in fatty-acid profiles between the tested organisms. Our results suggest that profiling fatty acids could be a useful approach in the taxonomic or phylogenetic study of the genus Nostoc and might serve as a valuable supplement to the current morphology-based classification system. PMID- 14756529 TI - Isolation and characterization of bacteria capable of degrading phenol and reducing nitrate under low-oxygen conditions. AB - Nitrate-reducing bacteria capable of degrading phenol were isolated from natural and contaminated environments under low-oxygen conditions with nitrate-containing media, using phenol as a sole carbon and energy source. A total of 27 bacteria able to degrade phenol and reduce nitrate under low-oxygen conditions were isolated from all of the inoculum samples, regardless of previous phenol contamination. For all of these bacteria, oxygen was an essential requirement for phenol degradation. Nitrate reduction by 19 of the strains was insensitive to 10 mM sodium azide, and these strains were placed into the alpha- and beta subclasses of Proteobacteria and two were Gram-positive bacteria. To date, the order of Rhizobiales has hardly been reported to have an ability to degrade aromatic compounds. Interestingly, our study showed that all isolates that were placed into the alpha-subclass of Proteobacteria are in the order of Rhizobiales. Furthermore, the genus Agrobacterium was isolated from most inoculum samples and one genus of Gram-positive bacteria, Staphylococcus, was also isolated. In the case of the remaining eight strains, nitrate reduction was inhibited by 10 mM sodium azide. Of these strains, seven were placed into the gamma-subclass of Proteobacteria. PMID- 14756531 TI - Cloning and characterization of the NAD-dependent 7alpha-Hydroxysteroid dehydrogenase from Bacteroides fragilis. AB - The NAD-linked 7alpha-hydroxysteroid dehydrogenase (7-HSDH) from Bacteroides fragilis ATCC 25285 was characterized and its gene cloned. The enzyme displayed optimal activities at pH 8.5 (NAD reduction) and 6.5 (NADH oxidation). The lowest K(m) and highest V(max) values were observed with chenodeoxycholic acid and its conjugates. The protein had subunits of 27.4 kDa and a native size of 110 kDa, suggesting a homotetrameric composition. The enzyme was relatively thermostable, retaining 95% of initial activity after 1 h at 65 degrees C. A DNA probe based on the N-terminal amino acid sequence hybridized to a 2373-bp HindIII fragment of B. fragilis DNA. This fragment was cloned into E. coli and sequenced, revealing a 780-bp open reading frame. The predicted amino acid sequence of the ORF showed strong sequence similarity to three other bacterial 7-HSDHs, all in the short chain dehydrogenase family. The regulation of expression of this gene is currently under investigation. PMID- 14756532 TI - Effect of Porphyromonas gingivalis vesicles on coaggregation of Staphylococcus aureus to oral microorganisms. AB - Vesicles from the outer membrane of Porphyromonas gingivalis have the ability to aggregate a wide range of Streptococcus spp., Fusobacterium nucleatum, Actinomyces naeslundii, and Actinomyces viscosus. We found that in the presence of P. gingivalis vesicles, Staphylococcus aureus coaggregated with Streptococcus spp., and the mycelium-type Candida albicans, but not the yeast type. Autoaggregation of S. aureus in the presence of P. gingivalis vesicles is inhibited by L-arginine, L-lysine, and L-cysteine. Both the methicillin-sensitive (MSSA) and -resistant (MRSA) strains of S. aureus were able to coaggregate with Streptococcus spp., A. naeslundii, and A. viscosus when they were treated with P. gingivalis vesicles. P. gingivalis vesicle-treated mycelium-type C. albicans coaggregated with S. aureus, but the yeast-type did not. These results indicate that strains of S. aureus, including MRSA, could adhere to oral biofilms in dental plaque on the tooth surface or in the gingival crevice when P. gingivalis is present. PMID- 14756533 TI - Protein profile of Acidithiobacillus ferrooxidans strains exhibiting different levels of tolerance to metal sulfates. AB - Strains of Acidithiobacillus ferrooxidans exhibited differences in the inhibition of Fe2+ oxidation in the presence of 250 mM of cadmium, zinc, and manganese sulfates in respirometric assays. Strains LR and 135 were practically not inhibited, whereas strains SSP and V3 showed significant inhibition (30-70%). Analysis by SDS-PAGE of total proteins from cells grown in the absence of metal sulfates showed different profiles between the more tolerant strains (LR and 135) and the more susceptible ones (SSP and V3). Total proteins of strains LR and V3 were also resolved by two-dimensional polyacrylamide gel electrophoresis (2-DE). A set of major proteins (40, 32, 22, and 20 kDa) could be identified only in the more tolerant strain LR. Our results show that protein profiles analysis could differentiate A. ferrooxidans strains that considerably differ in the tolerance to metal sulfates and present low genomic similarity as revealed by Random Amplified Polymorphic DNA (RAPD) data obtained previously in our laboratory. PMID- 14756534 TI - Unique distribution of GAL genes on chromosome XI in the yeast Saccharomyces naganishii. AB - A region of DNA extending from GAL7 to GAL1 was cloned in the yeast Saccharomyces naganishii. Sequence analysis revealed that GAL7 and GAL1 are separated by approximately 2 kbp and share a common promoter region. Although GAL7, GAL10, and GAL1 are clustered in this order in previously studied hemiascomycetous yeasts, GAL10 was not found between GAL7 and GAL1 in S. naganishii. Southern blotting of S. naganishii chromosomal DNA showed that both the GAL7-GAL1 region and GAL10 are located on chromosome XI, but that GAL10 is located more than 10 kbp away from GAL7-GAL1. Thus, S. naganishii and S. cerevisiae, while related phylogenetically, do not share the same orientation with respect to GAL genes. These data are highly relevant to studies of chromosomal evolution in yeast. PMID- 14756535 TI - Occurrence and linkage between secreted insecticidal toxins in natural isolates of Bacillus thuringiensis. AB - Little is known about the occurrence and linkage between secreted insecticidal virulence factors in natural populations of Bacillus thuringiensis (Bt). We carried out a survey of 392 Bt strains isolated from various samples originating from 31 countries. The toxicity profile of the culture supernatants of these strains was determined individually against Anthonomus grandis (Coleoptera) and Spodoptera littoralis (Lepidoptera). We analyzed beta-exotoxin I production and searched for the genes encoding Vip1-2, Vip3, and Cry1I toxins in 125 of these strains. Our results showed that these insecticidal toxins were widespread in Bt but that their distribution was nonrandom, with significant linkage observed between vip3 and cry1I and between vip1-2 and beta-exotoxin I. Strains producing significant amounts of beta-exotoxin I were more frequently isolated from invertebrate samples than from dust, water, soil, or plant samples. PMID- 14756536 TI - Survival of lactic acid bacteria in seawater: a factorial study. AB - A feasibility study of lactic bacteria as potential probiotics in larval cultures of marine fish was performed by investigating the survival of five strains of lactic bacteria in seawater by readily standardized procedures at different temperatures and salinities. These conditions were chosen in such a way that their combinations define a complete first-order factorial design. Depending on the strain and the ambient conditions, the survival adhered to first-order kinetics in some cases, and to the Gompertz equation in others. The half lives (t0.5) calculated from these models were subsequently introduced as responses to the factorial designs, estimating the coefficients of empirical equations that describe the group effect of temperature and salinity on t0.5. Simply additive effects were found in two cases, a negative first-order interaction in another case, while another two required second-order models. PMID- 14756537 TI - Effect of androgens and glucocorticoids on microbial growth and antimicrobial susceptibility. AB - The effects of androgens, testosterone and dihydrotestosterone (DHT), of an environmental anti-androgen, 2,2-bis(4-chlorophenyl)-1,1-dichloroethylene (DDE), and of glucocorticoids, hydrocortisone and dexamethasone, on growth kinetics and antibiotic susceptibility of E. faecalis, E. coli, P. aeurginosa, and S. aureus were measured. For P. aeurginosa, the presence of either DHT or DDE caused at least a fourfold shift in the minimum inhibitory concentration (MIC) of cefepime and tobramycin. DHT and DDE also affected the response of E. faecalis to meropenem and norfloxacin, resulting in a shift from sensitive to intermediate resistance (four-fold increase in MIC). Hydrocortisone (2 microM) induced an increase in the sensitivity of S. aureus to erythromycin, as compared to hormone free control (from 0.5 to 0.06 microg/mL). The susceptibility pattern of E. coli was unaffected by the hormones tested. These changes in susceptibility to antibiotics were unrelated to alterations in growth kinetics. For all organisms tested, the alterations in MICs occurred only in the presence of hormone, indicative of changes in the phenotype of these stable quality control strains. PMID- 14756539 TI - Prostate cancer screening policy in Italy. A statement endorsed by twenty scientific societies: towards unanimous consent. PMID- 14756538 TI - Adaptation of Pseudomonas fluorescens to Al-citrate: involvement of tricarboxylic acid and glyoxylate cycle enzymes and the influence of phosphate. AB - The degradation of Aluminum-citrate by Pseudomonas fluorescens necessitated a major restructuring of the various enzymatic activities involved in the TCA and glyoxylate cycles. While a six-fold increase in fumarase (FUM EC 4.2.1.2) activity was observed in cells subjected to Al-citrate compared to control cells, citrate synthase (CS EC 4.1.3.7) activity experienced a two-fold increase. On the other hand, in the Al-stressed cells malate synthase (MS EC 4.1.3.2) activity underwent a five-fold decrease in activity. This modulation of enzymatic activities appeared to be evoked by Al stress, as the incubation of Al-stressed cells in control media led to the complete reversal of these enzymatic profiles. These observations were further confirmed by 1H NMR and 13C NMR spectroscopy. No significant variations were observed in the activities of other glyoxylate and TCA cycle enzymes, like isocitrate lyase (ICL EC 4.1.3.1), malate dehydrogenase (MDH EC 1.1.1.37), and succinate dehydrogenase (SDH EC 1.3.99.1). This reconfiguration of the metabolic pathway appears to favour the production of a citrate-rich aluminophore that is involved in the sequestration of Al. PMID- 14756540 TI - Italian national consensus conference on prostate cancer screening (Florence, May 17, 2003)--final consensus document. PMID- 14756541 TI - Functional relationship and gene ontology classification of breast cancer biomarkers. AB - Breast cancer is a complex disease that still imposes a significant healthcare burden on women worldwide. The etiology of breast cancer is not known but significant advances have been made in the area of early detection and treatment. The advent of advanced molecular biology techniques, mapping of the human genome and availability of high throughput genomic and proteomic strategies opens up new opportunities and will potentially lead to the discovery of novel biomarkers for early detection and prognostication of breast cancer. Currently, many biomarkers, particularly the hormonal and epidermal growth factor receptors, are being utilized for breast cancer prognosis. Unfortunately, none of the biomarkers in use have sufficient diagnostic, prognostic and/or predictive power across all categories and stages of breast cancer. It is recognized that more useful information can be generated if tumors are interrogated with multiple markers. But choosing the right combination of biomarkers is challenging, because 1) multiple pathways are involved, 2) up to 62 genes and their protein products are potentially involved in breast cancer-related mechanisms and 3) the more markers evaluated, the more the time and cost involved. This review summarizes the current literature on selected biomarkers for breast cancer, discusses the functional relationships, and groups the selected genes based on a Gene Ontology classification. PMID- 14756542 TI - Prognostic value of S-phase fraction in 920 breast cancer patients: focus on T1N0 status. AB - The aim of this study was to reexamine the prognostic role of tumor cell kinetics measured by S-phase fraction (SPF) and to establish its clinically relevant threshold values. SPF was determined by flow cytometry in a group of 920 consecutive breast cancer patients, all followed at our institute for 10 years (1988 to 1998). Mean age was 60.5 years (27-89 years). Median follow-up was 63 months (3-150 months). All patients had initial surgical treatment. SPF quartiles were: Q1=3.08%, median value = 5.98%, Q3=10.22%. A significant difference in overall specific survival was obtained between two populations divided by a cutoff at Q1 (p < 0.0001). A multifactorial analysis including SPF and known prognostic factors such as tumor size, node status, histological grade, ER and PR status was performed using the Cox model in a population of 719 patients: univariate analysis showed that each of these factors had significant influence on overall survival. Multivariate analysis selected three of them, ranked by decreasing order of hazard ratio (HR) value: SPF (HR: 3.88, p < 0.001), tumor size (HR: 2.49, p < 0.001) and nodal status (HR: 2.28, p < 0.001). In addition, when tumors were stratified according to SPF quartile values, there were statistically different overall survival curves in patients with small tumors (< 2 cm) and in axillary node-negative patients. PMID- 14756543 TI - TP53 codon 72 polymorphism does not affect risk of cervical cancer in patients from The Gambia. AB - AIMS: A case-control study was performed to investigate the relationship between cervical cancer and TP53 polymorphism at codon 72 in young black African women from The Gambia. MATERIALS AND METHODS: The TP53 polymorphism at codon 72 was examined by PCR amplification and SSCP analysis in 40 patients with primary cervical cancer and in 20 healthy women of the same age and from the same geographical area. The occurrence of TP53 polymorphism in combination with the HPV-16 E6 genotype (assayed by PCR) was evaluated. RESULTS: The distribution of TP53 genotypes in cervical cancer patients and in the control group was not statistically different (p = 0.45) and homozygosity for argine at residue 72 was not associated with cervical cancer (odds ratio: 1.24; 95% confidence interval 0.21-9.16). Similarly, a different genotype distribution, cervical cancer and presence of HPV-16 E6 were not observed. CONCLUSIONS: These results cannot rule out an association between TP53 polymorphism at codon 72, HPV infection and the etiology of cervical cancer in this population sample. PMID- 14756545 TI - Analysis of bone alkaline phosphatase as a marker for the diagnosis of osteoporosis in men under androgen ablation. AB - The objective of this study was to evaluate the usefulness of serum determination of bone alkaline phosphatase (BAP) in the diagnosis of osteoporosis in men with prostate cancer under androgen ablation. Serum levels of BAP and bone mineral density (BMD) were assessed in 110 patients with non-metastatic, treated prostate cancer. Fifty-eight patients were under androgen deprivation during a period between two and 96 months and 52 had been submitted only to radical prostatectomy. Mean serum BAP was 11.8 ng/mL in patients with normal BMD, 16.7 ng/mL in patients with osteopenia (p. 0.058), and 19.3 ng/mL in patients with osteoporosis (p = 0.044). The correlation between serum BAP and BMD was significant (p. 0.006) but with an index of only 0.26. Receiver operating characteristic analysis for the diagnosis of osteoporosis showed an area under the curve of 0.608. None of the cutoff points that provided specificities of 75%, 90% and 95% gave significant distributions. The positive and negative predictive values as well as the odds ratios were not of any clinical usefulness. We conclude that serum BAP should not be considered a good marker for the diagnosis of osteoporosis in men with prostate cancer. Therefore, BAP serum determination cannot replace bone densitometry as a diagnostic tool. PMID- 14756544 TI - Prognostic value of p53 and MDM2 expression in bilharziasis-associated squamous cell carcinoma of the urinary bladder. AB - The aim of this study was to elucidate the associations between immunostaining for MDM2 and p53, their respective expression in squamous cell carcinoma of the urinary bladder, and the value of these variables for predicting treatment outcome after cystectomy. Inactivation of TP53 might play a role in the development and progression of bladder cancer. Complex formation with the MDM2 product is one mechanism that inactivates the p53 protein. Therefore, the MDM2 and the p53 protein were investigated to study potential interactions in bladder cancer. Fifty archival bladder tissue specimens were immunohistochemically stained using monoclonal antibodies against p53 and MDM2. Staining for p53 was observed in 48% of the specimens and staining for MDM2 in 20%. Univariate analysis demonstrated a significant correlation between p53 accumulation and survival (p = 0.0101), while the correlation between MDM2 and survival was not significant (p = 0.7183). The combined expression of MDM2 and p53 doest not add to the prognostic information provided by p53 alone. PMID- 14756546 TI - Serum tumor markers may precede instrumental response to chemotherapy in patients with metastatic cancer. AB - PURPOSE: Although serum tumor markers (STMs) are widely used in clinical practice, their predictive role for the response to anticancer treatment is still controversial. The correlation of CEA, CA 15.3, CA 19.9, CA 125 (only with peritoneal involvement) and NSE levels with imaging response and clinical benefit was investigated in 60 non-selected patients with metastatic epithelial cancers treated by single-agent docetaxel chemotherapy. METHODS: STM measurement was performed at baseline and subsequently every three to four weeks. We applied the WHO criteria to evaluate both STM and instrumental responses. Concordance analysis was performed by the Cohen Kw index, and the significance of the results was established using the Fleiss, Cohen & Everitt test. Qualitative interpretation of data was obtained with the Landis & Koch scale. Correlations of STM response with clinical benefit (PS or pain improvement) were evaluated by the chi-square test. RESULTS: The primary tumors included breast cancers (38 patients), gastrointestinal non-colorectal cancers (12 patients), and lung cancers (10 patients). An overall significant good degree of agreement was observed between STM and instrumental response (p < 0.0005). The degree of agreement for each marker was as follows: excellent for CEA (p < 0.0005) and CA 125 (p = 0.006), good for CA 15.3 (p < 0.0005) and CA 19.9 (p = 0.011). Restricted analysis for the correlation of each marker with primary tumor origin showed good prediction of radiological response for CA 15.3 and CEA in breast cancer patients (p<0.0005 for both), for CEA and CA 19.9 in gastrointestinal cancer patients (p = 0.01 and 0.04, respectively), and for CEA+NSE in lung cancer patients (p = 0.01). Conversely, STM response did not correlate significantly with the clinical benefit for the patients, both in terms of PS and pain improvement (p = 0.24 and p=0.42, respectively). CONCLUSION: This study showed STMs to be good predictors of tumor response. Although STMs cannot replace diagnostic imaging, in metastatic cancer they might be useful to optimize the timing of radiological re-evaluation in the palliative setting. PMID- 14756547 TI - Cytosolic levels of an estrogen-induced breast cancer-associated peptide (TFF1/pS2) in colorectal cancer: clinical significance and relationship with steroid receptors. AB - BACKGROUND: The Trefoil Factor 1 (TFF1/pS2), a peptide consisting of 60 amino acids, is the most abundant estrogen-induced messenger RNA in MCF-7 breast cancer cells and is also expressed by colorectal carcinomas. The objective of this work was to evaluate the cytosolic TFF1 content in colorectal carcinomas, its possible relationship with estrogen and progesterone receptors as well as with clinicopathological tumor parameters, and its potential prognostic significance. METHODS: Cytosolic TFF1 levels were examined by immunoradiometric assay in 178 patients with resectable colorectal cancer. The mean follow-up period was 32 months. RESULTS: There was a wide variability of cytosolic TFF1 levels in tumor surrounding mucosa samples (0.09-42.5 ng/mg protein) as well as in tumors (0.01 270 ng/mg protein). Comparison of paired mucosa and carcinoma samples showed significantly higher TFF1 levels in tumors (mean: 17.1 ng/mg protein) than in mucosa samples (10 ng/mg protein) (p = 0.027). TFF1 levels were significantly higher in mucosa samples surrounding distal colon and rectal tumors (p = 0.0001) and in tumor samples obtained from older patients (p = 0.007). However, there were no significant differences in tumor TFF1 levels with respect to clinicopathological parameters such as the patient's sex, tumor location, stage, histological grade, ploidy, S-phase, or tumor estrogen and progesterone receptors. In addition, there was no significant relationship between tumor TFF1 levels and disease outcome. CONCLUSIONS: TFF1 may play an as yet undetermined role in the tumorigenesis of colorectal carcinomas. However, cytosolic levels of TFF1 do not seem to have any prognostic significance in colorectal carcinomas. PMID- 14756548 TI - Serum IL-13 concentration response to surgical treatment in patients with laryngeal squamous cell carcinoma. PMID- 14756549 TI - Structural insights into chloride and proton-mediated gating of CLC chloride channels. AB - CLC Cl(-) channels fulfill numerous physiological functions as demonstrated by their involvement in several human genetic diseases. They have an unusual homodimeric architecture in which each subunit forms an individual pore whose open probability is regulated by various physicochemical factors, including voltage, Cl(-) concentration, and pH. The voltage dependence of Torpedo channel CLC-0 is derived probably indirectly from the translocation of a Cl(-) ion through the pore during the opening step. Recent structure determinations of bacterial CLC homologues marked a breakthrough for the structure-function analysis of CLC channels. The structures revealed a complex fold with 18 alpha helices and two Cl(-) ions per subunit bound in the center of the protein. The side chain of a highly conserved glutamate residue that resides in the putative permeation pathway appears to be a major component of the channel gate. First studies have begun to exploit the bacterial structures as guides for a rational structure-function analysis. These studies confirm that the overall structure seems to be conserved from bacteria to humans. A full understanding of the mechanisms of gating of eukaryotic CLC channels is, however, still lacking. PMID- 14756550 TI - Hydrophobic core fluidity of homologous protein domains: relation of side-chain dynamics to core composition and packing. AB - The side-chain dynamics of methyl groups in two structurally related proteins from the fibronectin type III (fnIII) superfamily, the third fnIII domain from human tenascin (TNfn3) and the tenth fnIII domain from human fibronectin (FNfn10), have been studied by NMR spectroscopy. Side-chain order parameters reveal that the hydrophobic cores of the two proteins have substantially different mobilities. The core of TNfn3 is very dynamic, with exceptionally low order parameters for the most deeply buried residues, while that of FNfn10 is more like those of other proteins which have been studied with this technique, having a relatively rigid core with uniformly distributed dynamics. The unusually dynamic core of TNfn3 appears to be related to its amino acid composition, which makes it more fluid-like. A further explanation for the mobility of the TNfn3 core may be found in the negative correlation between the order parameter and excess packing volume, which shows that the core of TNfn3 is less densely packed and consequently has lower methyl order parameters for its buried residues. Rotameric transitions, presumably facilitated by the lower packing density, appear to make an important contribution to lowering the order parameters, and have been probed by measuring three-bond scalar couplings. Overall, although backbone dynamics is generally similar for proteins with the same topology on a fast time scale (picoseconds to nanoseconds), this study shows that a single fold can accommodate a wide variation in the dynamic properties of its core. PMID- 14756551 TI - Crystal structure of sucrose phosphorylase from Bifidobacterium adolescentis. AB - Around 80 enzymes are implicated in the generic starch and sucrose pathways. One of these enzymes is sucrose phosphorylase, which reversibly catalyzes the conversion of sucrose and orthophosphate to d-Fructose and alpha-d-glucose 1 phosphate. Here, we present the crystal structure of sucrose phosphorylase from Bifidobacterium adolescentis (BiSP) refined at 1.77 A resolution. It represents the first 3D structure of a sucrose phosphorylase and is the first structure of a phosphate-dependent enzyme from the glycoside hydrolase family 13. The structure of BiSP is composed of the four domains A, B, B', and C. Domain A comprises the (beta/alpha)(8)-barrel common to family 13. The catalytic active-site residues (Asp192 and Glu232) are located at the tips of beta-sheets 4 and 5 in the (beta/alpha)(8)-barrel, as required for family 13 members. The topology of the B' domain disfavors oligosaccharide binding and reduces the size of the substrate access channel compared to other family 13 members, underlining the role of this domain in modulating the function of these enzymes. It is remarkable that the fold of the C domain is not observed in any other known hydrolases of family 13. BiSP was found as a homodimer in the crystal, and a dimer contact surface area of 960 A(2) per monomer was calculated. The majority of the interactions are confined to the two B domains, but interactions between the loop 8 regions of the two barrels are also observed. This results in a large cavity in the dimer, including the entrance to the two active sites. PMID- 14756552 TI - Structural basis for the exocellulase activity of the cellobiohydrolase CbhA from Clostridium thermocellum. AB - Numerous bacterial and fungal organisms have evolved elaborate sets of modular glycoside hydrolases and similar enzymes aimed at the degradation of polymeric carbohydrates. Presently, on the basis of sequence similarity catalytic modules of these enzymes have been classified into 90 families. Representatives of a particular family display similar fold and catalytic mechanisms. However, within families distinctions occur with regard to enzymatic properties and type of activity against carbohydrate chains. Cellobiohydrolase CbhA from Clostridium thermocellum is a large seven-modular enzyme with a catalytic module belonging to family 9. In contrast to other representatives of that family possessing only endo- and, in few cases, endo/exo-cellulase activities, CbhA is exclusively an exocellulase. The crystal structures of the combination of the immunoglobulin like module and the catalytic module of CbhA (Ig-GH9_CbhA) and that of an inactive mutant Ig-GH9_CbhA(E795Q) in complex with cellotetraose (CTT) are reported here. The detailed analysis of these structures reveals that, while key catalytic residues and overall fold are conserved in this enzyme and those of other family 9 glycoside hydrolases, the active site of GH9_CbhA is blocked off after the -2 subsite. This feature which is created by an extension and altered conformation of a single loop region explains the inability of the active site of CbhA to accommodate a long cellulose chain and to cut it internally. This altered loop region is responsible for the exocellulolytic activity of the enzyme. PMID- 14756553 TI - Structural insights into the human and avian IMP cyclohydrolase mechanism via crystal structures with the bound XMP inhibitor. AB - Within de novo purine biosynthesis, the AICAR transformylase and IMP cyclohydrolase activities of the bifunctional enzyme ATIC convert the intermediate AICAR to the final product of the pathway, IMP. Identification of the AICAR transformylase active site and a proposed formyl transfer mechanism have already resulted from analysis of crystal structures of avian ATIC in complex with substrate and/or inhibitors. Herein, we focus on the IMPCH active site and the cyclohydrolase mechanism through comparison of crystal structures of XMP inhibitor complexes of human ATIC at 1.9 A resolution with the previously determined avian enzyme. This first human ATIC structure was also determined to ascertain whether any subtle structural differences, compared to the homologous avian enzyme, should be taken into account for structure-based inhibitor design. These structural comparisons, as well as comparative analyses with other IMP and XMP binding proteins, have enabled a catalytic mechanism to be formulated. The primary role of the IMPCH active site appears to be to induce a reconfiguration of the substrate FAICAR to a less energetically favorable, but more reactive, conformer. Backbone (Arg64 and Lys66) and side chain interactions (Thr67) in the IMPCH active site reorient the 4-carboxamide from the preferred conformer that binds to the AICAR Tfase active site to one that promotes intramolecular cyclization. Other backbone amides (Ile126 and Gly127) create an oxyanion hole that helps orient the formyl group for nucleophilic attack by the 4-carboxamide amine and then stabilize the anionic intermediate. Several other residues, including Lys66, Tyr104, Asp125, and Lys137', provide substrate specificity and likely enhance the catalytic rate through contributions to acid-base catalysis. PMID- 14756554 TI - Catalytic mechanism of the cyclohydrolase activity of human aminoimidazole carboxamide ribonucleotide formyltransferase/inosine monophosphate cyclohydrolase. AB - The bifunctional enzyme aminoimidazole carboxamide ribonucleotide transformylase/inosine monophosphate cyclohydrolase (ATIC) is responsible for catalysis of the last two steps in the de novo purine pathway. Using recently determined crystal structures of ATIC as a guide, four candidate residues, Lys66, Tyr104, Asp125, and Lys137, were identified for site-directed mutagenesis to study the cyclohydrolase activity of this bifunctional enzyme. Steady-state kinetic experiments on these mutants have shown that none of these residues are absolutely required for catalytic activity; however, they strongly influence the efficiency of the reaction. Since the FAICAR binding site is made up mostly of backbone interactions with highly conserved residues, we postulate that these conserved interactions orient FAICAR in the active site to favor the intramolecular ring closure reaction and that this reaction may be catalyzed by an orbital steering mechanism. Furthermore, it was shown that Lys137 is responsible for the increase in cyclohydrolase activity for dimeric ATIC, which was reported previously by our laboratory. From the experiments presented here, a catalytic mechanism for the cyclohydrolase activity is postulated. PMID- 14756555 TI - Three-dimensional structure of kynureninase from Pseudomonas fluorescens. AB - Kynureninase [E.C. 3.7.1.3] is a pyridoxal-5'-phosphate (PLP)-dependent enzyme that catalyzes the hydrolytic cleavage of l-kynurenine to anthranilic acid and l alanine. Sequence alignment with other PLP-dependent enzymes indicated that kynureninase is in subgroup IVa of the aminotransferases, along with nifS, CsdB, and serine-pyruvate aminotransferase, which suggests that kynureninase has an aminotransferase fold. Crystals of Pseudomonas fluorescens kynureninase were obtained, and the structure was solved by molecular replacement using the CsdB coordinates combined with multiple isomorphous heavy atom replacement. The coordinates were deposited in the PDB (ID code 1QZ9). The structure, refined to an R factor of 15.5% to 1.85 A resolution, is dimeric and has the aminotransferase fold. The structure also confirms the prediction from sequence alignment that Lys-227 is the PLP-binding residue in P. fluorescens kynureninase. The conserved Asp-201, expected for an aminotransferase fold, is located near the PLP nitrogen, but Asp-132 is also strictly conserved and at a similar distance from the pyridinium nitrogen. Mutagenesis of both conserved aspartic acids shows that both contribute equally to PLP binding, but Asp-201 has a greater role in catalysis. The structure shows that Tyr-226 donates a hydrogen bond to the phosphate of PLP. Unusual among PLP-dependent enzymes, Trp-256, which is also strictly conserved in kynureninases from bacteria to humans, donates a hydrogen bond to the phosphate through the indole N1-hydrogen. PMID- 14756556 TI - Transition state structure for ADP-ribosylation of eukaryotic elongation factor 2 catalyzed by diphtheria toxin. AB - Bacterial protein toxins are the most powerful human poisons known, exhibiting an LD(50) of 0.1-1 ng kg(-)(1). A major subset of such toxins is the NAD(+) dependent ADP-ribosylating exotoxins, which include pertussis, cholera, and diphtheria toxin. Diphtheria toxin catalyzes the ADP ribosylation of the diphthamide residue of eukaryotic elongation factor 2 (eEF-2). The transition state of ADP ribosylation catalyzed by diphtheria toxin has been characterized by measuring a family of kinetic isotope effects using (3)H-, (14)C-, and (15)N labeled NAD(+) with purified yeast eEF-2. Isotope trapping experiments yield a commitment to catalysis of 0.24 at saturating eEF-2 concentrations, resulting in suppression of the intrinsic isotope effects. Following correction for the commitment factor, intrinsic primary kinetic isotope effects of 1.055 +/- 0.003 and 1.022 +/- 0.004 were observed for [1(N)'-(14)C]- and [1(N)-(15)N]NAD(+), respectively; the double primary isotope effect was 1.066 +/- 0.004 for [1(N)' (14)C, 1(N)-(15)N]NAD(+). Secondary kinetic isotope effects of 1.194 +/- 0.002, 1.101 +/- 0.003, 1.013 +/- 0.005, and 0.988 +/- 0.002 were determined for [1(N)' (3)H]-, [2(N)'-(3)H]-, [4(N)'-(3)H]-, and [5(N)'-(3)H]NAD(+), respectively. The transition state structure was modeled using density functional theory (B1LYP/6 31+G) as implemented in Gaussian 98, and theoretical kinetic isotope effects were subsequently calculated using Isoeff 98. Constraints were varied in a systematic manner until the calculated kinetic isotope effects matched the intrinsic isotope effects. The transition state model most consistent with the intrinsic isotope effects is characterized by the substantial loss in bond order of the nicotinamide leaving group (bond order = 0.18, 1.99 A) and weak participation of the attacking imidazole nucleophile (bond order = 0.03, 2.58 A). The transition state structure imparts strong oxacarbenium ion character to the ribose ring even though significant bond order remains to the nicotinamide leaving group. The transition state model presented here is asymmetric and consistent with a dissociative S(N)1 type mechanism in which attack of the diphthamide nucleophile lags behind departure of the nicotinamide. PMID- 14756557 TI - Conserved and nonconserved residues in the substrate binding site of 7,8 diaminopelargonic acid synthase from Escherichia coli are essential for catalysis. AB - The vitamin B(6)-dependent enzyme 7,8-diaminopelargonic acid (DAPA) synthase catalyzes the antepenultimate step in the synthesis of biotin, the transfer of the alpha-amino group of S-adenosyl-l-methionine (SAM) to 7-keto-8 aminopelargonic acid (KAPA) to form DAPA. The Y17F, Y144F, and D147N mutations in the active site were constructed independently. The k(max)/K(m)(app) values for the half-reaction with DAPA of the Y17F and Y144F mutants are reduced by 1300- and 2900-fold, respectively, compared to the WT enzyme. Crystallographic analyses of these mutants do not show significant changes in the structure of the active site. The kinetic deficiencies, together with a structural model of the enzyme PLP/DAPA Michaelis complex, point to a role of these two residues in recognition of the DAPA/KAPA substrates and in catalysis. The k(max)/K(m)(app) values for the half-reaction with SAM are similar to that of the WT enzyme, showing that the two tyrosine residues are not involved in this half-reaction. Mutations of the conserved Arg253 uniquely affect the SAM kinetics, thus establishing this position as part of the SAM binding site. The D147N mutant is catalytically inactive in both half-reactions. The structure of this mutant exhibits significant changes in the active site, indicating that this residue plays an important structural role. Of the four residues examined, only Tyr144 and Arg253 are strictly conserved in the available amino acid sequences of DAPA synthases. This enzyme thus provides an illustrative example that active site residues essential for catalysis are not necessarily conserved, i.e., that during evolution alternative solutions for efficient catalysis by the same enzyme arose. Decarboxylated SAM [S-adenosyl-(5')-3-methylthiopropylamine] reacts nearly as well as SAM and cannot be eliminated as a putative in vivo amino donor. PMID- 14756558 TI - Similar molecular interactions of factor VII and factor VIIa with the tissue factor region that allosterically regulates enzyme activity. AB - Tissue factor (TF) binds the zymogen (VII) and activated (VIIa) forms of coagulation factor VII with high affinity. The structure determined for the sTF VIIa complex [Banner, D. W., et al. (1996) Nature 380, 41-46] shows that all four domains of VIIa (Gla, EGF-1, EGF-2, and protease) are in contact with TF. Although a structure is not available for the TF-VII complex, the structure determined for free VII [Eigenbrot, C., et al. (2001) Structure 9, 675-682] suggests a significant conformational change for the zymogen to enzyme transition. In particular, the region of the protease domain that must contact TF has a conformation that is altered from that of VIIa, suggesting that the VII protease domain interacts with TF in a manner different from that of VIIa. To test this hypothesis, a panel of 12 single-site sTF mutants, having substitutions of residues observed to contact the proteolytic domain of VIIa, have been evaluated for binding to both zymogen VII and VIIa. Affinities were determined by surface plasmon resonance measurements using a noninterfering anti-TF monoclonal antibody to capture TF on the sensor chip surface. Dissociation constants (K(D)) measured for binding to wild-type sTF are 7.5 +/- 2.4 nM for VII and 5.1 +/- 2.3 nM for VIIa. All of the sTF mutants except S39A and E95A exhibited a significant decrease (>2-fold) in affinity for VIIa. The changes in affinity measured for VII or VIIa binding with substitution in sTF were comparable in magnitude. We conclude that the proteolytic domain of both VII and VIIa interacts with this region of sTF in a nearly identical fashion. Therefore, zymogen VII can readily adopt a VIIa-like conformation required for binding to TF. PMID- 14756559 TI - Crystallographic studies on structural features that determine the enzymatic specificity and potency of human angiogenin: Thr44, Thr80, and residues 38-41. AB - Human angiogenin (Ang) is a potent inducer of blood vessel formation and is a member of the pancreatic ribonuclease superfamily. Its enzymatic activity is unusually weak and biased toward cleavage after cytidine nucleotides. As part of an ongoing investigation into the structural basis of Ang's characteristic activity, we have determined the crystal structures of three Ang variants having novel activity. (i) The structure of T44D-Ang indicates that Asp44 can participate directly in pyrimidine binding and that the intrinsic hydrogen bonding capability of this residue largely governs the pyrimidine specificity of this variant. Unexpectedly, the mutation also causes the most extensive disruption of the C-terminus seen in any Ang variant thus far. This allows the side chain of Arg101 to penetrate the B(1) site, raising the possibility that it participates in substrate binding as occurs in ribonuclease 4. (ii) The structure of T80A-Ang supports the view that Thr80 plays little role in maintaining the obstructive conformation of the C-terminus and that its participation in a hydrogen bond with Thr44 selectively weakens the interaction between Thr44 and N3 of cytosine. (iii) ARH-II is an angiogenin/RNase A chimera in which residues 38 41 of Ang are replaced with the corresponding residues (38-42) of RNase A. Its structure suggests that the guest segment influences catalysis by subtle means, possibly by reducing the pK(a) of the catalytic lysine. The loss of angiogenic activity is not attributable to disruption of known cell-binding or nuclear translocation sites but may be a consequence of the chimera's enhanced ribonucleolytic activity. PMID- 14756560 TI - Trypsin disrupts the trafficking of the human dopamine transporter by alpha synuclein and its A30P mutant. AB - Alpha-synuclein modulates dopamine homeostasis in dopamine-producing neurons of substantia nigra, partly through regulation of human dopamine transporter (hDAT) activity. To identify the underlying mechanisms, we disrupted the modulation of hDAT activity by wild-type (wt) alpha-synuclein, and its familial Parkinson's disease linked mutants A30P and A53T, by mild trypsinization (0.1%, 30 s) of Ltk( ) cotransfected cells. Trypsin completely reversed the attenuation of hDAT function mediated by wt and the A30P mutant. In A53T coexpressing cells, where DAT activity is not downregulated, trypsinization did not induce any changes. These effects of trypsin were mimicked by collagenase I and Dispase (0.1%, 1 min each) but not by chymotrypsin, Pronase, or papain (0.1%, up to 2 min each). Trypsin increased dopamine uptake in rat primary mesencephalic neurons, suggesting that DAT activity is also subjected to modulation by alpha-synuclein in these neurons that endogenously coexpress both proteins. In trypsinized cells, dopamine accelerated both production of reactive oxygen species and cell death in hDAT and wt or A30P, but not A53T, coexpressing cells, compared to nontrypsinized cells. Paradoxically, trypsin increased the protein-protein interactions between the synuclein variants and hDAT, without any noticeable proteolysis of these proteins. hDAT-alpha-synuclein protein-protein interactions occurred through residues 58-107 (NAC domain) of the alpha-synuclein variants and residues 598-620 of the carboxy-terminal tail of hDAT, in both trypsinized and nontrypsinized cells. Confocal microscopy and biotinylation studies show that, in cells expressing the wt or A30P variants, but not the A53T mutant, hDAT is sequestered away from the plasma membrane into the cytoplasm, an effect that is reversed by trypsin. These results show that alpha-synuclein modulates hDAT function through trafficking of the transporter in a process that can be disrupted by trypsin. PMID- 14756561 TI - Pseudosubstrate peptides inhibit Akt and induce cell growth inhibition. AB - We have designed peptide inhibitors that potently inhibit Akt both in vitro and inside cells. These peptide inhibitors are selective for Akt versus other closely related kinases. The peptides inhibit the in vitro phosphorylation of a biotinylated Bad peptide by Akt with potency up to 100 nM. We have shown that the binding between Akt1 and these peptide inhibitors requires MgATP. Mutating the two putative Akt phosphorylation sites to Ala (nonsubstrate) in these peptides increases the inhibitory potency while mutating the sites to aspartic acid (phosphorylation mimetic) reduces the potency. When delivered into cells, these peptide inhibitors can inhibit cellular Akt activity and cell growth. Thus, these Akt-specific peptide inhibitors provide prototypes for peptide mimetic drugs as well as very useful tools to dissect cellular functions of Akt. PMID- 14756562 TI - Removal of PsaF alters forward electron transfer in photosystem I: evidence for fast reoxidation of QK-A in subunit deletion mutants of Synechococcus sp. PCC 7002. AB - Recent studies of point mutations in photosystem I have suggested that the two kinetic phases of phylloquinone reoxidation represent electron transfer in the two branches of cofactors. This interpretation implies that changes in the relative amplitudes of the two kinetic phases represent a change in the extent of electron transfer in the two branches. Using time-resolved electron paramagnetic resonance (EPR), this issue is investigated in subunit deletion mutants of Synechococcus sp. PCC 7002. The spin-polarized EPR signals of P(700)(+)A(1)(-) and P(700)(+)FeS(-), both at room temperature and in frozen solution, are altered by deletion of PsaF and/or PsaE, and the differences from the wild type are much more pronounced in PS I complexes isolated from the mutants using Triton X-100 rather than n-dodecyl beta-d-maltopyranoside. The changes in the transient EPR data for the mutant complexes are consistent with a significant fraction of reaction centers showing (i) faster electron transfer from A(1)(-) to F(X), (ii) slower forward electron transfer from A(0)(-) to A(1), and (iii) slightly altered quinone hyperfine couplings, possibly as a result of a change in the hydrogen bonding. The fraction of fast electron transfer and its dependence on the isolation procedure are estimated approximately from simulations of the room temperature EPR data. The results are discussed in terms of possible models for the electron transfer. It is suggested that the detergent-induced fraction of fast electron transfer is most likely due to alteration of the environment of the quinone in the PsaA branch of cofactors and is not the result of a change in the directionality of electron transfer. PMID- 14756563 TI - Hydrophobic pockets at the membrane interface: an original mechanism for membrane protein interactions. AB - The effect of partial digestion by trypsin and GluC protease on the association of the membrane polypeptides of LH1 from Rhodospirillum (Rsp.) rubrum was studied. Trypsin and GluC protease treatments of LH1 result in the cleavage of the first three amino acids from the alpha polypeptide and of the first 18 amino acids from the beta polypeptide, respectively, without any noticeable reorganization of their secondary structure, as measured by attenuated total reflectance Fourier transform IR spectroscopy. However, the enthalpy variation accompanying dimer formation was dramatically reduced by the protease attacks by as much as 80%. Our results show that the alphabeta heterodimer is mainly stabilized by hydrophobic interactions which involve the amino-terminal extensions of the participating polypeptides. Using the close homology between the polypeptides of Rsp. rubrum LH1 and that of Rsp. molischianum LH2, whose structure is known, a structural model for these "hydrophobic pockets" lying close to the membrane interface is proposed. PMID- 14756564 TI - Conformational changes in the rod domain of human keratin 8 following heterotypic association with keratin 18 and its implication for filament stability. AB - Keratin intermediate filaments are heteropolymers of type I and type II polypeptides that constitute the bulk of the epithelial cytoskeleton. We microinjected seven keratin monoclonal antibodies into human epithelial cells, and two of them, only A45-B/B3 and LP3K, caused the formation of keratin aggregates. The keratin filaments in human epithelial cells were also disrupted by a monovalent A45-B/B3 Fab fragment, suggesting that the binding of the antibody, rather than cross-linking, collapses the filaments. Immunoblotting and ELISA experiments suggested that the antibody reacted weakly with recombinant K8 but did not react with recombinant K18 at all. However, the antibody reactivity increased substantially when a mixture of the two keratin polypeptides, either recombinant or derived from MCF-7, was used. The epitopes of 15 monoclonal antibodies recognizing human K8 were characterized by their reactivity with recombinant fragments of K8. Reactivity of antibody A45-B/B3 with fragments of K8 in the presence of K18 revealed that the antibody recognizes an epitope in the rod domain of K8, between residues 313 and 332, on the amino-terminal side of the stutter in helix 2B, which is involved in heterotypic association. The data suggest that this region of K8 undergoes a conformational change following interaction with the complementary K18 either to expose the epitope or to increase its affinity for the antibody. Taken together, the data highlight the role of this epitope in heterotypic association and in filament stabilization. PMID- 14756565 TI - Relocation or duplication of the helix A sequence of T4 lysozyme causes only modest changes in structure but can increase or decrease the rate of folding. AB - In T4 lysozyme, helix A is located at the amino terminus of the sequence but is associated with the C-terminal domain in the folded structure. To investigate the implications of this arrangement for the folding of the protein, we first created a circularly permuted variant with a new amino terminus at residue 12. In effect, this moves the sequence corresponding to helix A from the N- to the C-terminus of the molecule. The protein crystallized nonisomorphously with the wild type but has a very similar structure, showing that the unit consisting of helix A and the C-terminal domain can be reconstituted from a contiguous polypeptide chain. The protein is less stable than the wild type but folds slightly faster. We then produced a second variant in which the helix A sequence was appended at the C terminus (as in the first variant), but was also restored at the N-terminus (as in the wild type). This variant has two helix A sequences, one at the N-terminus and the other at the C-terminus, each of which can compete for the same site in the folded protein. The crystal structure shows that it is the N-terminal sequence that folds in a manner similar to that of the wild type, whereas the copy at the C-terminus is forced to loop out. The stability of this protein is much closer to that of the wild type, but its rate of folding is significantly slower. The reduction in rate is attributed to the presence of the two identical sequence segments which compete for a single, mutually exclusive, site. PMID- 14756567 TI - HMGB6 from Arabidopsis thaliana specifies a novel type of plant chromosomal HMGB protein. AB - The high-mobility group (HMG) proteins of the HMGB family are chromatin associated proteins that act as architectural factors in various nucleoprotein structures, which regulate DNA-dependent processes such as transcription and recombination. Database analyses revealed that in addition to the previously identified HMGB1-HMGB5 proteins, the Arabidopsis genome encodes at least three other family members having the typical overall structure of a central HMG-box DNA binding domain, which is flanked by basic and acidic regions. These novel HMGB proteins display some structural differences, when compared to HMGB1-HMGB5. Therefore, a representative of the identified proteins, now termed HMGB6, was further analyzed. The HMGB6 protein of approximately 27 kDa is the largest plant HMGB protein identified so far. This is essentially due to its unusually extended N-terminal domain of 109 amino acid residues. Subcellular localization experiments demonstrate that it is a nuclear protein. According to CD measurements, HMGB6 has an alpha-helical HMG-box domain. HMGB6 can bind DNA structure-specifically, and it is a substrate for the protein kinase CK2alpha. Because of these features, HMGB6, and presumably its relatives, can be considered members of the plant HMGB protein family. Hence, eight different chromosomal HMGB proteins are expressed in Arabidopsis, and they may serve specialized architectural functions assisting various DNA-dependent processes. PMID- 14756566 TI - Lysine directed cross-linking of viral DNA-RNA:DNA hybrid substrate to the isolated RNase H domain of HIV-1 reverse transcriptase. AB - An isolated ribonuclease H domain of HIV-1 reverse transcriptase is capable of specifically removing the tRNA primer within an oligonucleotide mimic. The determinants for substrate specificity are located in a region within the terminal octanucleotide of the acceptor stem of the tRNA. Recognition of the substrate by HIV-1 RNase H was analyzed by the introduction of a cross-linking reagent directed toward lysines on the thymine residue complementary to the scissile bond, facing the major groove of the DNA-RNA:DNA substrate. Cross linking of the modified substrate to RNase H required the presence of Mn(2+). The Mn(2+) titration of cross-linking paralleled the Mn(2+) requirement for activity. Modified substrate quenched with glycine prior to binding of substrate was efficiently cleaved, whereas the RNA within the cross-linked product was intact. Tryptic digestion of the isolated RNase H-nucleic acid covalent complex revealed a main cross-linked peptide whose N-terminal peptide sequence is VVTLTDTTNQ, indicating that the cross-linked lysine corresponds to Lys476. Cross-linking to K476 was confirmed by analysis of K476C RNase H. Mutation of K476C disrupted the chemical cross-linking while maintaining activity. On the basis of the size of the cross-linker arm, the results indicate that K476 is in closer proximity to the tRNA mimic substrate within the isolated RNase H domain than observed for the RNase H-resistant polypurine tract (PPT) substrate within the HIV-1 RT. PMID- 14756568 TI - Binding of bovine pancreatic trypsin inhibitor to trypsinogen: spectroscopic and volumetric studies. AB - We have investigated the binding of bovine pancreatic trypsin inhibitor (BPTI) to bovine trypsinogen by combining ultrasonic velocimetry, high precision densimetry, and fluorescence spectroscopy. We report the changes in volume, adiabatic compressibility, van't Hoff enthalpy, entropy, and free energy that accompany the association of the two proteins at 25 degrees C and pH 8.0. We have used the measured changes in volume and compressibility in conjunction with available structural data to characterize the binding-induced changes in the hydration properties and intrinsic packing of the two proteins. Our estimate reveals that 110 +/- 40 water molecules become released to the bulk from the hydration shells of BPTI and trypsinogen. Furthermore, we find that the intrinsic coefficient of adiabatic compressibility of the two proteins decreases by 14 +/- 2%, which is suggestive of the binding-induced rigidification of the proteins' interior. BPTI-trypsinogen association is an entropy-driven event which proceeds with an unfavorable change in enthalpy. The favorable change in entropy results from partial compensation between two predominant terms. Namely, a large favorable change in hydrational entropy slightly prevails over a close in magnitude but opposite in sign change in configurational entropy. The reduction in configurational entropy and, consequently, protein dynamics is consistent with the observed decrease in intrinsic compressibility. In general, results of this work emphasize the vital role that water plays in modulating protein recognition events. PMID- 14756569 TI - Enzymatic mechanism of low-activity mouse alcohol dehydrogenase 2. AB - ADH2 is a member of one of the six classes of mammalian alcohol dehydrogenases, which catalyze the reversible oxidation of alcohols using NAD(+) as a cofactor. Within the ADH2 class, the rodent enzymes form a subgroup that exhibits low catalytic activity with all substrates that were examined, as compared to other groups, such as human ADH2. The low activity can be ascribed to the rigid nature of the proline residue at position 47 as the activity can be increased by approximately 100-fold by substituting Pro47 with either His (as found in human ADH2), Ala, or Gln. Mouse ADH2 follows an ordered bi-bi mechanism, and hydride transfer is rate-limiting for oxidation of benzyl alcohols catalyzed by the mutated and wild-type enzymes. Structural studies suggest that the mouse enzyme with His47 has a more closed active site, as compared to the enzyme with Pro47, and hydride transfer can be more efficient. Oxidation of benzyl alcohol catalyzed by all forms of the enzyme is strongly pH dependent, with pK values in the range of 8.1-9.3 for turnover numbers and catalytic efficiency. These pK values probably correspond to the ionization of the zinc-bound water or alcohol. The pK values are not lowered by the Pro47 to His substitution, suggesting that His47 does not act as a catalytic base in the deprotonation of the zinc ligand. PMID- 14756570 TI - Additive transfer free energies of the peptide backbone unit that are independent of the model compound and the choice of concentration scale. AB - With knowledge of individual transfer free energies of chemical groups that become newly exposed on protein denaturation and assuming the group transfer free energy contributions are additive, it should be possible to predict the stability of a protein in the presence of denaturant. Unfortunately, several unresolved issues have seriously hampered quantitative development of this transfer model for protein folding/unfolding. These issues include the lack of adequate demonstration that group transfer free energies (DeltaG(tr)) are additive and independent of the choice of model compound, the problem arising from dependence of DeltaG(tr) on concentration scales, the lack of knowledge of activity coefficients, and the validity of the mathematical constructs used in obtaining DeltaG(tr) values. Regarding transfer from water to 1 M concentrations of the naturally occurring osmolytes, trimethylamine-N-oxide (TMAO), sarcosine, betaine, proline, glycerol, sorbitol, sucrose, trehalose, and urea, using cyclic glycylglycine, zwitterionic glycine peptides, and N-acetylglycine amide peptides as models for the peptide backbone of proteins, we set out to address these issues and obtain DeltaG(tr) values for the peptide backbone unit. We demonstrate experimental approaches that obviate the choice of concentration scale and demonstrate additivity in DeltaG(tr) of the peptide backbone unit for all solvent systems studied. Evidence is presented to show that the DeltaG(tr) values are independent of the chemical model studied, and experimental conditions are given to illustrate when the mathematical constructs are valid and when activity coefficients can be ignored. Resolution of the long-standing issues that have stymied development of the transfer model now make it possible to design transfer experiments that yield reliable and quantitative values for the interactions between osmolyte-containing solvents and native and unfolded protein. PMID- 14756571 TI - Importance of clustered 2'-O-(2-aminoethyl) residues for the gene targeting activity of triple helix-forming oligonucleotides. AB - We are developing triple helix-forming oligonucleotides (TFOs) as gene targeting reagents in living mammalian cells. We have described psoralen-linked TFOs with 2'-O-methyl and 2'-O-(2-aminoethyl) (2'-AE) substitutions that are active in a gene knockout assay in cultured cells. The assay is based on mutagenesis by psoralen, a photoactive DNA cross-linker. Previous work showed that TFOs with three or four 2'-AE residues were disproportionately more active than those with one or two substitutions. Here we demonstrate that for optimal bioactivity the 2' AE residues must be clustered rather than dispersed. We have further characterized bioactive and inactive TFOs in an effort to identify biochemical and biophysical correlates of biological activity. While thermal stability is a standard monitor of TFO biophysical activity, we find that T(m) values do not distinguish bioactive and inactive TFOs. In contrast, measurements of TFO association rates appear to correlate well with bioactivity, in that triplex formation occurs disproportionately faster with the TFOs containing three or four 2'-AE residues. We asked if extending the incubation time prior to photoactivation would enhance the bioactivity of a TFO with a slow on rate relative to the TFO with a faster association rate. However, there was no change in bioactivity differential. These results are compatible with a model in which TFO binding in vivo is followed by relatively rapid elution by cellular functions, similar to that described for transcription factors. Under these circumstances, TFOs with faster on rates would be favored because they would be more likely to be in triplexes at the time of photoactivation. PMID- 14756572 TI - Structural characterization of macroH2A containing chromatin. AB - MacroH2A (mH2A) is one of the most recently identified members of the heteromorphous histone variant family. It is unique among the members of this group because it contains an unusually large non-histone C-terminal end, from where its name derives, and appears to be restricted to subphylum vertebrata. Although a concerted effort has been carried out in order to characterize the physiological relevance of mH2A, little is known in comparison about the structural importance of the molecule. Elucidating the biophysical and conformational proprieties of mH2A in chromatin may provide clues into the links between this histone variant and its unique function(s). In this paper, we look first at the heterogeneous tissue-specific distribution of this protein in different vertebrate classes. This is followed by a structural comparison between mH2A and H2A protein and by the characterization of the nucleosome core particles with which these histone subtypes are associated. We find that the highly alpha helical C-terminus of mH2A confers an asymmetric conformation to nucleosomes and that this variant is tightly bound to chromatin fragments in a way that does not depend on the overall extent of acetylation of the other core histones. PMID- 14756573 TI - The side chain of aspartic acid 69 dictates the folding mechanism of Bacillus subtilis HPr. AB - Many small, single-domain proteins show equilibrium and kinetic folding mechanisms that appear to be adequately described as two state. The two-state model makes several predictions that can be tested experimentally. First, the conformational stability determined at or extrapolated to a set of reference conditions should be independent of the measurement method (thermal or solvent denaturation or hydrogen exchange). Second, model-independent measures of the cardinal thermodynamic parameters (T(m), DeltaH) as determined from direct calorimetric means should be identical to those determined from the two-state analysis of thermal unfolding data. Third, the ratio of the kinetic folding and unfolding rate constants should be equal to K(eq) determined from an equilibrium measurement under the same conditions. Here, we show that the wild-type HPr protein from Bacillus subtilis does not meet all of these criteria under our standard conditions. However, if we replace the side chain of Asp69, or add moderate concentrations of salt, we find excellent two-state behavior in both equilibrium and kinetic folding. Thus, for this protein and possibly others, very subtle changes in the primary structure or in the solution conditions can dramatically alter the relative stabilities of the native intermediate, and unfolded ensembles can cause an observable change in the nature of the folding mechanism. PMID- 14756574 TI - Bidirectional electron transfer in photosystem I: accumulation of A0- in A-side or B-side mutants of the axial ligand to chlorophyll A0. AB - Photosystem I contains two potential electron transfer pathways between P(700) and F(X). These branches are made up of the electron transfer chain components A, A(0), and A(1). The primary electron acceptor A(0) is a chlorophyll a monomer that could be one or both of the two chlorophyll molecules, eC-A(3)/eC-B(3), identified in the 2.5 A resolution structure. The eC-A(3)/eC-B(3) chlorophylls are both coordinated by the sulfur atom of a methionine. This coordination is highly unusual, as interactions between the acid Mg(2+) and the soft base sulfur are weak. The eC-A(3)/eC-B(3) chlorophylls also are located close to one of the connecting chlorophylls that may link the antenna and the electron transfer chain chlorophylls. Due to their location in the structure, the eC-A(3)/eC-B(3) chlorophylls may play a role in both excitation energy transfer and electron transfer. To test the role of the eC-A(3)/eC-B(3) chlorophylls in electron transfer, Met-684 of PsaA and Met-664 of PsaB have been changed to His, Ser, and Leu. Replacement of either M(A684) or M(B664) results in a significant alteration in growth phenotype. The His and Leu mutants are very light sensitive in the presence of oxygen. Growth is impaired to a greater extent in the B-side mutants. However, all of the mutants are able to grow anaerobically at comparable rates. The His and Ser mutants all accumulate PSI at a level similar to that of wild type, whereas the Leu mutants have reduced amounts of PSI. Ultrafast transient absorbance measurements show that the (A(0)(-) - A(0)) difference signal accumulates in the MH(A684) and MH(B664) mutants under neutral conditions, demonstrating that electron transfer between A(0)(-) and A(1) is blocked or significantly slowed. The results show that both the A-branch and the B-branch of the ETC are active in PSI from Chlamydomonas reinhardtii. PMID- 14756575 TI - Substrate binding and reduction of benzoyl-CoA reductase: evidence for nucleotide dependent conformational changes. AB - Benzoyl-CoA reductase (BCR) from the denitrifying bacterium Thauera aromatica catalyzes the ATP driven two-electron reduction of the aromatic moiety of benzoyl CoA (BCoA) to a nonaromatic cyclic diene (2 ATP/2 e(-)). The enzyme contains two similar but nonidentical ATP-binding sites of the acetate kinase/sugar kinase/Hsp70/actin family. To obtain further insights into the overall catalytic cycle of BCR, the binding affinities and stoichiometries of all substrates as well as their effects on reduction kinetics were studied by stopped-flow UV/vis spectroscopy, freeze-quench EPR spectroscopy, and equilibrium dialysis. BCR bound maximally two nucleotides and a single BCoA. The binding of a single nucleotide induced a molecular switch (BCR --> BCR) as evidenced as follows: (i) the reduction rate of BCR by sulfoxide radical anion was significantly decreased in the nucleotide-bound state, (ii) the binding of BCoA to the reduced enzyme strictly depended on bound nucleotides, and (iii) the nucleotide binding affinities increased up to 60-fold compared to the steady-state values. The "ATP binding switch" is distinguished from the previously described "low-spin/high spin switch" of a [4Fe-4S] cluster which strictly depends on ATP hydrolysis. The two nucleotide binding sites were occupied sequentially; binding constants of the two sites differed by a factor of 10-40. The kinetic data obtained suggest that the ATP-binding switch is a rather fast process (>100 s(-)(1)) with a switch equilibrium constant of 54 +/- 10. In contrast, the reverse switch back of the MgADP-bound enzyme (BCR --> BCR) is considered rate-limiting in the overall catalytic cycle of BCR (4 +/- 1 s(-)(1)). The binding of nucleotides did not affect the redox potential of the [4Fe-4S](+1/+2) clusters; the switch is rather considered to alter the kinetics of internal electron transfer. Implications for the overall catalytic cycle of benzoyl-CoA reductase are discussed and compared with other ATP-hydrolyzing enzymes. PMID- 14756576 TI - FhuF, part of a siderophore-reductase system. AB - FhuF is a cytoplasmic 2Fe-2S protein of Escherichia coli loosely associated with the cytoplasmic membrane. E. coli fhuF mutants showed reduced growth on plates with ferrioxamine B as the sole iron source, although siderophore uptake was not defective in transport experiments. Removal of iron from coprogen, ferrichrome, and ferrioxamine B was significantly lower in fhuF mutants compared to the corresponding parental strains, which suggested that FhuF is involved in iron removal from these hydroxamate-type siderophores. A redox potential E(1/2) of 310 +/- 25 mV relative to the normal hydrogen electrode was determined for FhuF by EPR redox titration; this redox potential is sufficient to reduce the siderophores coprogen and ferrichrome. Mossbauer spectra revealed that FhuF in its [Fe(2+)-Fe(3+)] state is also capable of direct reduction of ferrioxamine B bound ferric iron, thus proving its reductase function. This is the first report on a bacterial siderophore-iron reductase which in vivo seems to be specific for a certain group of hydroxamates. PMID- 14756577 TI - Selective antagonism to the cadherin BT-R1 interferes with calcium-induced adhesion of epithelial membrane vesicles. AB - BT-R(1) is a member of the cadherin superfamily of proteins and is expressed in the midgut epithelium of Manduca sexta during larval development. Previously, we showed that calcium ions influence the structure and stability of BT-R(1) on brush border membrane vesicles (BBMVs) prepared from M. sexta midgut epithelium. In the present study, the effects of calcium and Cry1Ab toxin, produced by Bacillus thuringiensis, on the adhesive properties of BBMVs were investigated. Addition of calcium to a suspension of BBMVs promoted adhesion and aggregation of the vesicles. Treatment of BBMVs with trypsin or lowering the pH (pH 4.0) of the BBMV suspension abolished calcium-induced vesicle aggregation, whereas treatment with deglycosylating enzymes did not affect the aggregation of vesicles, indicating that adhesion and clustering of BBMVs involves protein-protein interactions. Preincubation of BBMVs with Cry1Ab toxin, which specifically binds to BT-R(1) with high affinity and disrupts the midgut epithelium of M. sexta, caused a 50% decrease in calcium-induced vesicle aggregation. The inhibitory effects of the Cry1Ab toxin on BBMV aggregation was blocked completely when the toxin was preincubated with a peptide containing the toxin-binding site of BT R(1). Cry3A toxin, which is similar in molecular structure to Cry1Ab but does not bind to BT-R(1) and is not toxic to M. sexta larvae, did not affect BBMV aggregation. The results of this study demonstrate that the adhesive function of BT-R(1) is compromised by the Cry1Ab toxin, which acts as a selective antagonist, and supports the notion that BT-R(1) is critical in preserving the integrity of larval midgut epithelium in M. sexta. PMID- 14756579 TI - Safety and tolerability of lamotrigine for bipolar disorder. AB - Tolerability and safety are important considerations in optimising pharmacotherapy for bipolar disorder. This paper reviews the tolerability and safety of lamotrigine, an anticonvulsant recommended in the 2002 American Psychiatric Association guidelines as a first-line treatment for acute depression in bipolar disorder and one of several options for maintenance therapy. This paper reviews the tolerability and safety of lamotrigine using data available from a large programme of eight placebo-controlled clinical trials of lamotrigine enrolling a total of nearly 1800 patients with bipolar disorder. This review is the first to collate all the safety information from these clinical trials, including data from four unpublished studies. The results these trials in which 827 patients with bipolar disorder were given lamotrigine as monotherapy or adjunctive therapy for up to 18 months for a total of 280 patient-years of exposure demonstrated that lamotrigine is well-tolerated with an adverse-event profile generally comparable with that of placebo. The most common adverse event with lamotrigine was headache. Lamotrigine did not appear to destabilise mood and was not associated with sexual adverse effects, weight gain, or withdrawal symptoms. Few patients experienced serious adverse events with lamotrigine, and the incidence of withdrawals because of adverse events was low. Serious rash occurred rarely (0.1% incidence) in the clinical development programme including both controlled and uncontrolled clinical trials. These findings - considered in the context of data showing lamotrigine to be effective for bipolar depression - establish lamotrigine as a well-tolerated addition to the psychotropic armamentarium. PMID- 14756578 TI - Pharmacogenetic aspects of drug-induced torsade de pointes: potential tool for improving clinical drug development and prescribing. AB - Drug-induced torsade de pointes (TdP) has proved to be a significant iatro-genic cause of morbidity and mortality and a major reason for the withdrawal of a number of drugs from the market in recent times. Enzymes that metabolise many of these drugs and the potassium channels that are responsible for cardiac repolarisation display genetic polymorphisms. Anecdotal reports have suggested that in many cases of drug-induced TdP, there may be a concealed genetic defect of either these enzymes or the potassium channels, giving rise to either high plasma drug concentrations or diminished cardiac repolarisation reserve, respectively. The presence of either of these genetic defects may predispose a patient to TdP, a potentially fatal adverse reaction, even at therapeutic dosages of QT-prolonging drugs and in the absence of other risk factors. Advances in pharmacogenetics of drug metabolising enzymes and pharmacological targets, together with the prospects of rapid and inexpensive genotyping procedures, promise to individualise and improve the benefit/risk ratio of therapy with drugs that have the potential to cause TdP. The qualitative and the quantitative contributions of these genetic defects in clinical cases of TdP are unclear because not all of the patients with TdP are routinely genotyped and some relevant genetic mutations still remain to be discovered. There are regulatory guidelines that recommend strategies aimed at uncovering the risk of TdP associated with new chemical entities during their development. There are also a number of guidelines that recommend integrating pharmacogenetics in this process. This paper proposes a strategy for integrating pharmacogenetics into drug development programmes to optimise association studies correlating genetic traits and endpoints of clinical interest, namely failure of efficacy or development of repolarisation abnormalities. Until pharmacogenetics is carefully integrated into all phases of development of QT-prolonging drugs and large-scale studies are undertaken during their post-marketing use to determine the genetic components involved in induction of TdP, routine genotyping of patients remains unrealistic. Even without this pharmacogenetic data, the clinical risk of TdP can already be greatly minimised. Clinically, a substantial proportion of cases of TdP are due to the use of either high or usual dosages of drugs with potential to cause TdP in the presence of factors that inhibit drug metabolism. Therefore, choosing the lowest effective dose and identifying patients with these non-genetic risk factors are important means of minimising the risk of TdP. In view of the common secondary pharmacology shared by these drugs, a standard set of contraindications and warnings have evolved over the last decade. These include factors responsible for pharmacokinetic or pharmacodynamic drug interactions. Among the latter, the more important ones are bradycardia, electrolyte imbalance, cardiac disease and co-administration of two or more QT-prolonging drugs. In principle, if large scale prospective studies can demonstrate a substantial genetic component, pharmacogenetically driven prescribing ought to reduce the risk further. However, any potential benefits of pharmacogenetics will be squandered without any reduction in the clinical risk of TdP if physicians do not follow prescribing and monitoring recommendations. PMID- 14756580 TI - Benefit-risk assessment of rofecoxib in the treatment of osteoarthritis. AB - NSAIDs are widely used to treat pain and inflammation in osteoarthritis. Their use in this indication is generally intermittent and fluctuates with the intensity of the disease. Nonetheless, success of the therapy is frequently limited by injury to the gastrointestinal mucosa and complications such as bleeding, ulceration and perforation. A careful and detailed evaluation of these aspects in regard to the newly introduced NSAIDs is of considerable clinical importance. This review focuses on the NSAID rofecoxib, one of the selective cyclo-oxygenase (COX)-2 inhibitors, which are claimed to be as effective as nonselective NSAIDs with better gastrointestinal tolerability. Indeed, phase II, phase III and epidemiological studies have revealed that the efficacy of rofecoxib is comparable to that of conventional NSAIDs but with lower gastrointestinal toxicity, although this advantage may not be demonstrable in every patient. In patients treated with low-dose aspirin (acetylsalicylic acid) for cardiovascular prophylaxis, celecoxib (another selective COX-2 inhibitor) seems to have no obvious advantages over conventional NSAIDs, and similar conclusions may be applied to rofecoxib. A comparison of NSAID therapy +/- concomitant low-dose aspirin was not a primary outcome in this trial with celecoxib and there is thus a need for further studies which compare the gastrointestinal risk of a selective COX-2 inhibitor plus aspirin versus a conventional NSAID. Recent debate has emerged regarding the cardiovascular safety of rofecoxib. Although there is evidence both for and against higher cardiovascular risk with rofecoxib, a retrospective cohort study recently published suggested that there is no increased risk of acute myocardial infarction in the short-term when compared with non-selective NSAIDs. The renal toxicity of rofecoxib has been thoroughly investigated. Clinical studies revealed renal effects of rofecoxib similar to those of conventional NSAIDs. Since adverse effects increase with the degree of renal impairment, monitoring of renal function should be carried out in patients at risk. Although there are still insufficient data concerning certain important adverse effects of rofecoxib, this drug is becoming an important alternative in the therapy of osteoarthritis, especially in high-risk patients. Clinicians need to weigh up the benefits and risks of rofecoxib on a case-by-base basis. PMID- 14756581 TI - Malformation rates in children of women with untreated epilepsy: a meta-analysis. AB - BACKGROUND: It is widely quoted that women with epilepsy have a higher than baseline risk for giving birth to a child with malformations, independent of the effects of antiepileptic drugs. OBJECTIVE: To determine, based on available evidence, if epilepsy per se represents a teratogenic risk. To systematically review all studies investigating the occurrence of major malformation rates among children of treated or untreated women with epilepsy and non-exposed controls who do not have epilepsy. METHODS: A meta-analysis, using a random effects model, was conducted of all cohort and case-control studies reporting malformation rates in children of women with epilepsy exposed or unexposed to antiepileptic drugs compared with that of children of nonepileptic women. Medline (1966-2001), EMBASE, the Cochrane database as well as REPROTOX (an information system on environmental hazards to human reproduction and development) databases were accessed. RESULTS: We found ten studies reporting results of untreated epilepsy (n = 400) and their non-epileptic healthy controls (n = 2492). Nine out of ten studies also reported results on 1443 patients exposed to antiepileptic drugs and their 2526 unexposed healthy controls. The risk for congenital malformations in the offspring of women with untreated epilepsy was not higher than among nonepileptic controls (odds ratio [OR] = 1.92; 95% CI 0.92-4.00). There was evidence of publication bias, thus with bias removed the OR was 0.99 (95% CI 0.49 2.01). In contrast, the offspring of epileptic women who received antiepileptic drugs had higher incidences of malformation than controls (OR 3.26; 95% CI 2.15 4.93). CONCLUSION: Our study does not support the commonly held view that epilepsy per se represents a teratogenic risk. Our study suggests that this view is the result of a publication bias, with several small (< 100 participants) positive studies leading to a premature conclusion. PMID- 14756582 TI - The risk of severe depression, psychosis or panic attacks with prophylactic antimalarials. AB - INTRODUCTION/OBJECTIVE: Experimental and observational studies have linked mefloquine use to an increased risk of developing neuropsychiatric adverse effects such as depression or psychoses. Most of these reports relied on interview-based information from travellers. We conducted a population-based observational study using a database of medical records to quantify and compare the risk of psychiatric disorders during or after use of mefloquine with the risk during use of proguanil and/or chloroquine, or doxycycline. STUDY DESIGN/METHODS: The study population was drawn from the large UK-based General Practice Research Database (GPRD). Subjects were aged from 17-79 years and were exposed to mefloquine, proguanil, chloroquine or doxycycline (or a combination of these drugs) at some time between 1990 and 1999. We performed a person-time and a nested case-control analysis to assess the risk of developing a first-time diagnosis of depression, psychosis or panic attack during or after use of these antimalarial drugs. RESULTS: Within the study population of 35 370 subjects (45.2% males), we identified 580 subjects with a first-time diagnosis of depression (n = 505), psychosis (n = 16) or panic attack (n = 57) and two subjects committed suicide. The incidence rates of first-time diagnoses of depression during current use of mefloquine, proguanil and/or chloroquine, or doxycycline, adjusted for age, gender and calendar year, were 6.9 (95% CI 4.5 10.6), 7.6 (95% CI 5.5-10.5) and 9.5 (95% CI 3.7-24.1)/1000 person-years, respectively. The incidence rates of psychosis or panic attacks during current mefloquine exposure were 1.0/1000 person-years (95% CI 0.3-2.9) and 3.0/1000 person-years (95% CI 1.6-5.7), respectively, approximately 2-fold higher (statistically nonsignificant) than during current use of proguanil and/or chloroquine, or doxycycline. The nested case-control analysis encompassed 505 cases with depression and 3026 controls, 16 cases with psychosis and 96 controls, and 57 cases with a panic attack and 342 controls. Current use of mefloquine was not associated with an elevated risk of developing depression. In a comparison between patients currently using mefloquine with all past users of antimalarials combined, the risk estimate was elevated for current users of mefloquine for both psychosis (odds ratio [OR] 8.0, 95% CI 1.0-62.7; p < 0.05) and panic attacks (OR 2.7, 95% CI 1.1-6.5; p < 0.05). CONCLUSION: The absolute risk of developing psychosis or panic attack appears low with all the antimalarials tested. No evidence was found in this large observational study that mefloquine use increased the risk of first-time diagnosis of depression when compared with the use of other antimalarials investigated in this study. PMID- 14756583 TI - A theory of causal learning in children: causal maps and Bayes nets. AB - The authors outline a cognitive and computational account of causal learning in children. They propose that children use specialized cognitive systems that allow them to recover an accurate "causal map" of the world: an abstract, coherent, learned representation of the causal relations among events. This kind of knowledge can be perspicuously understood in terms of the formalism of directed graphical causal models, or Bayes nets. Children's causal learning and inference may involve computations similar to those for learning causal Bayes nets and for predicting with them. Experimental results suggest that 2- to 4-year-old children construct new causal maps and that their learning is consistent with the Bayes net formalism. PMID- 14756584 TI - Addiction motivation reformulated: an affective processing model of negative reinforcement. AB - This article offers a reformulation of the negative reinforcement model of drug addiction and proposes that the escape and avoidance of negative affect is the prepotent motive for addictive drug use. The authors posit that negative affect is the motivational core of the withdrawal syndrome and argue that, through repeated cycles of drug use and withdrawal, addicted organisms learn to detect interoceptive cues of negative affect preconsciously. Thus, the motivational basis of much drug use is opaque and tends not to reflect cognitive control. When either stressors or abstinence causes negative affect to grow and enter consciousness, increasing negative affect biases information processing in ways that promote renewed drug administration. After explicating their model, the authors address previous critiques of negative reinforcement models in light of their reformulation and review predictions generated by their model. PMID- 14756585 TI - A detection-theoretic model of echo inhibition. AB - A detection-theoretic analysis of the auditory localization of dual-impulse stimuli is described, and a model for the processing of spatial cues in the echo pulse is developed. Although for over 50 years "echo suppression" has been the topic of intense theoretical and empirical study within the hearing sciences, only a rudimentary understanding of its mechanisms has emerged. In this article, psychometric functions and results from matching studies are used in developing a model that specifies the perceived position of the echo pulse as a normal deviate, with an expectation that is a logistic function of the echo delay and a variance that depends on interaural time difference. Loss of information in the echo event is quantified as a decline in the efficiency with which the binaural system receives information from the lag impulse. PMID- 14756586 TI - Spiking Phineas Gage: a neurocomputational theory of cognitive-affective integration in decision making. AB - The authors present a neurological theory of how cognitive information and emotional information are integrated in the nucleus accumbens during effective decision making. They describe how the nucleus accumbens acts as a gateway to integrate cognitive information from the ventromedial prefrontal cortex and the hippocampus with emotional information from the amygdala. The authors have modeled this integration by a network of spiking artificial neurons organized into separate areas and used this computational model to simulate 2 kinds of cognitive-affective integration. The model simulates successful performance by people with normal cognitive-affective integration. The model also simulates the historical case of Phineas Gage as well as subsequent patients whose ability to make decisions became impeded by damage to the ventromedial prefrontal cortex. PMID- 14756587 TI - Modeling the effects of choice-set size on the processing of letters and words. AB - Letters and words are better identified when there are fewer available choices. How do readers use choice-set restrictions? By analyzing new experimental data and previously reported data, the author shows that Bayes theorem-based models overestimate readers' use of choice-set restrictions. This result is discordant with choice-similarity models such as R. D. Luce's (1963a) similarity choice model, G. Keren and S. Baggen's (1981) letter recognition model, and D. W. Massaro and G. C. Oden's (1979) fuzzy logical model of perception. Other models posit that choice restrictions affect accuracy only by improving guessing (e.g., J. L. McClelland & D. E. Rumelhart's, 1981, interactive activation model). It is shown that these models underestimate readers' use of choice-set restrictions. Restriction of choice set does improve perception of letters and words, but not optimally. Decision models that may be able to explain this phenomenon are discussed. PMID- 14756588 TI - Reconceptualizing individual differences in self-enhancement bias: an interpersonal approach. AB - Self-enhancement bias has been studied from 2 perspectives: L. Festinger's (1954) social comparison theory (self-enhancers perceive themselves more positively than they perceive others) and G. W. Allport's (1937) self-insight theory (self enhancers perceive themselves more positively than they are perceived by others). These 2 perspectives are theoretically and empirically distinct, and the failure to recognize their differences has led to a protracted debate. A new interpersonal approach to self-enhancement decomposes self-perception into 3 components: perceiver effect, target effect, and unique self-perception. Both theoretical derivations and an illustrative study suggest that this resulting measure of self-enhancement is less confounded by unwanted components of interpersonal perception than previous social comparison and self-insight measures. Findings help reconcile conflicting views about whether self enhancement is adaptive or maladaptive. PMID- 14756589 TI - Obsessive-compulsive disorder as a disturbance of security motivation. AB - The authors hypothesize that the symptoms of obsessive-compulsive disorder (OCD), despite their apparent nonrationality, have what might be termed an epistemic origin--that is, they stem from an inability to generate the normal "feeling of knowing" that would otherwise signal task completion and terminate the expression of a security motivational system. The authors compare their satiety-signal construct, which they term yedasentience, to various other senses of the feeling of knowing and indicate why OCD-like symptoms would stem from the abnormal absence of such a terminator emotion. In addition, they advance a tentative neuropsychological model to explain its underpinnings. The proposed model integrates many previous disparate observations and concepts about OCD and embeds it within the broader understanding of normal motivation. PMID- 14756590 TI - The timing of meals. AB - In most individuals, food intake occurs as discrete bouts or meals, and little attention has been paid to the factors that normally determine when meals will occur when food is freely available. On the basis of experiments using rats, the authors suggest that when there are no constraints on obtaining food and few competing activities, 3 levels of interacting controls normally dictate when meals will start. The first is the genetically determined circadian activity pattern on which nocturnal animals tend to initiate most meals in the dark. The second is the regularly occurring changing of the light cycle: These changes provide temporal anchors. The third relates to the size of the preceding meal, such that larger meals cause a longer delay until the onset of the next meal. Superimposed on these 3 are factors related to learning, convenience, and opportunity. PMID- 14756591 TI - Perceiving the intensity of light. AB - The relationship between luminance (i.e., the photometric intensity of light) and its perception (i.e., sensations of lightness or brightness) has long been a puzzle. In addition to the mystery of why these perceptual qualities do not scale with luminance in any simple way, "illusions" such as simultaneous brightness contrast, Mach bands, Craik-O'Brien-Cornsweet edge effects, and the Chubb Sperling-Solomon illusion have all generated much interest but no generally accepted explanation. The authors review evidence that the full range of this perceptual phenomenology can be rationalized in terms of an empirical theory of vision. The implication of these observations is that perceptions of lightness and brightness are generated according to the probability distributions of the possible sources of luminance values in stimuli that are inevitably ambiguous. PMID- 14756592 TI - A diffusion model account of the lexical decision task. AB - The diffusion model for 2-choice decisions (R. Ratcliff, 1978) was applied to data from lexical decision experiments in which word frequency, proportion of high- versus low-frequency words, and type of nonword were manipulated. The model gave a good account of all of the dependent variables--accuracy, correct and error response times, and their distributions--and provided a description of how the component processes involved in the lexical decision task were affected by experimental variables. All of the variables investigated affected the rate at which information was accumulated from the stimuli--called drift rate in the model. The different drift rates observed for the various classes of stimuli can all be explained by a 2-dimensional signal-detection representation of stimulus information. The authors discuss how this representation and the diffusion model's decision process might be integrated with current models of lexical access. PMID- 14756593 TI - The architecture of personality. AB - This article presents a theoretical framework for analyzing psychological systems that contribute to the variability, consistency, and cross-situational coherence of personality functioning. In the proposed knowledge-and-appraisal personality architecture (KAPA), personality structures and processes are delineated by combining 2 principles: distinctions (a) between knowledge structures and appraisal processes and (b) among intentional cognitions with varying directions of fit, with the latter distinction differentiating among beliefs, evaluative standards, and aims. Basic principles of knowledge activation and use illuminate relations between knowledge and appraisal, yielding a synthetic account of personality structures and processes. Novel empirical data illustrate the heuristic value of the knowledge/appraisal distinction by showing how self referent and situational knowledge combine to foster cross-situational coherence in appraisals of self-efficacy. PMID- 14756594 TI - Structure and deterioration of semantic memory: a neuropsychological and computational investigation. AB - Wernicke (1900, as cited in G. H. Eggert, 1977) suggested that semantic knowledge arises from the interaction of perceptual representations of objects and words. The authors present a parallel distributed processing implementation of this theory, in which semantic representations emerge from mechanisms that acquire the mappings between visual representations of objects and their verbal descriptions. To test the theory, they trained the model to associate names, verbal descriptions, and visual representations of objects. When its inputs and outputs are constructed to capture aspects of structure apparent in attribute-norming experiments, the model provides an intuitive account of semantic task performance. The authors then used the model to understand the structure of impaired performance in patients with selective and progressive impairments of conceptual knowledge. Data from 4 well-known semantic tasks revealed consistent patterns that find a ready explanation in the model. The relationship between the model and related theories of semantic representation is discussed. PMID- 14756595 TI - On the composition of risk preference and belief. AB - Prospect theory assumes nonadditive decision weights for preferences over risky gambles. Such decision weights generalize additive probabilities. This article proposes a decomposition of decision weights into a component reflecting risk attitude and a new component depending on belief. The decomposition is based on an observable preference condition and does not use other empirical primitives such as statements of judged probabilities. The preference condition is confirmed by most of the experimental findings in the literature. The implied properties of the belief component suggest that, besides the often-studied ambiguity aversion (a motivational factor reflecting a general aversion to unknown probabilities), perceptual and cognitive limitations play a role: It is harder to distinguish among various levels of likelihood, and to process them differently, when probabilities are unknown than when they are known. PMID- 14756596 TI - Holography does not account for goodness: a critique of van der Helm and Leeuwenberg (1996). AB - P. A. van der Helm and E. L. J. Leeuwenberg (1996) outlined a holographic account of figural goodness of a perceptual stimulus. The theory is mathematically precise and can be applied to a broad spectrum of empirical data. The authors argue, however, that the account is inadequate on both theoretical and empirical grounds. The theoretical difficulties concern the internal consistency of the account and its reliance on unspecified auxiliary assumptions. The account also makes counterintuitive empirical predictions, which do not fit past data or the results of a series of new experimental studies. PMID- 14756598 TI - Psychological and physical adjustment to breast cancer over 4 years: identifying distinct trajectories of change. AB - The goal of this study was to identify distinct trajectories of adjustment to breast cancer over 4 years as well as to distinguish among the different trajectories. The mental and physical functioning of 287 women with breast cancer who remained alive and disease free through 4 years of follow-up were examined. The majority of women showed slight and steady improvement in functioning with time, but subgroups of women were identified who showed marked improvement and marked deteriorations over time. Age successfully distinguished different trajectories of physical functioning. Indices of personal resources (i.e., self image, optimism, perceived control) and social resources (i.e., social support) successfully distinguished different courses of mental and physical functioning. PMID- 14756599 TI - Is finding something good in the bad always good? Benefit finding among women with breast cancer. AB - The correlates and consequences of benefit finding on quality of life were examined for 364 women (93% Caucasian, 6% African American, and 1% Hispanic) diagnosed with Stage I, II, and III breast cancer. Benefit finding and quality of life were measured 4 months postdiagnosis (Tl), 3 months after Tl (T2), and 6 months after T2 (T3). Women with lower socioeconomic status, minorities, and those with more severe disease perceived more benefits at baseline. Benefit finding was associated with more negative affect at baseline and also interacted with stage of disease, such that negative relations to quality of life across time were limited to those with more severe disease. Findings suggest there are qualifiers as to whether "finding something good in the bad" is good or bad. PMID- 14756600 TI - Experiences of demand and control in daily life as correlates of subclinical carotid atherosclerosis in a healthy older sample. AB - Daily experiences of demand and control were examined as correlates of carotid artery atherosclerosis among healthy adults (ages 50-70). Mediating effects of ambulatory blood pressure (ABP) were also explored. Participants (n=337) collected ABP and recorded daily experiences, using electronic diaries, over two 3-day periods. Carotid artery intima-medial thickness (IMT) was assessed using ultrasonography. Participants reporting higher task demands during daily life showed larger IMT, after adjustment for demographic covariates. This association was not limited to workplace ratings or to employed individuals. The association was mediated, in part, by daytime systolic blood pressure. Previous findings linking job stress with cardiovascular disease may reflect the broader impact of daily psychological demands, not necessarily associated with the workplace. PMID- 14756601 TI - Understanding and predicting parental decisions about early childhood immunizations. AB - This research investigated the factors that influence decisions about immunizations. Women in the third trimester of pregnancy (N=195) rated their likelihood of immunizing their child; stated their reasons for and against immunizing; and rated their perceptions of the benefits and risks of immunization, feelings of responsibility, and anticipated regret if harm occurred. Immunization status was determined at follow-up. Stepwise regression analyses demonstrated that immunization decisions are strongly influenced by omission bias factors such as anticipated responsibility and regret variance (which explained more than 50% of variance). It is suggested that parents may benefit from antenatal decision aids that address omission bias and encourage them to assess benefits and risks of immunizations on the basis of scientific evidence. PMID- 14756602 TI - Coping and anxiety in women recalled for additional diagnostic procedures following an abnormal screening mammogram. AB - This study characterized women's concurrent and subsequent levels of emotional distress associated with a questionable mammogram screening and relationships between women's coping and psychosocial adjustment. State anxiety was assessed in 98 women 1 day after receiving a mammogram screening (Time 1), after notification of a questionable screening result that necessitated additional testing (Time 2), and after being informed of their breast-cancer-free status (Time 3). Key findings include (a) women reported a significant increase in anxiety following notification of the need to return for follow-up testing; (b) significant and positive associations were found between anxiety and behavioral approach, behavioral avoidance, cognitive approach, and cognitive avoidance coping in cross sectional analyses; and (c) cognitive avoidance coping was a strong predictor of final levels of state anxiety in women. Findings suggest that cognitive avoidance coping plays an important role in reducing anxiety in women recalled to clarify an initially ambiguous screening procedure. PMID- 14756603 TI - Pain catastrophizing is associated with health indices in musculoskeletal pain: a cross-sectional study in the Dutch community. AB - Cross-sectional associations were examined between pain catastrophizing and several health indices in 1,164 people with musculoskeletal pain from a Dutch community sample. Health indices included in the present study were specialist consultation, use of medication, and absenteeism or work disability. The results demonstrate that for people with a current episode of musculoskeletal pain, pain catastrophizing, pain intensity, and the presence of multiple pain locations were significantly associated with specialist consultation, use of pain medication, and absenteeism or work disability. The authors conclude that the role of pain itself has perhaps been underestimated in recent models of chronic pain-related disability. Some clinical implications and suggestions for further research are given. PMID- 14756604 TI - Testing a self-determination theory process model for promoting glycemic control through diabetes self-management. AB - A longitudinal study tested the self-determination theory (SDT) process model of health behavior change for glycemic control within a randomized trial of patient activation versus passive education. Glycosylated hemoglobin for patients with Type 2 diabetes (n=159) was assessed at baseline, 6 months, and 12 months. Autonomous motivation and perceived competence were assessed at baseline and 6 months, and the autonomy supportiveness of clinical practitioners was assessed at 3 months. Perceptions of autonomy and competence were promoted by perceived autonomy support, and changes in perceptions of autonomy and competence, in turn, predicted change in glycemic control. Self-management behaviors mediated the relation between change in perceived competence and change in glycemic control. The self-determination process model fit the data well. PMID- 14756605 TI - The impact of rape on women's sexual health risk behaviors. AB - This study used cluster analysis to identify three patterns of sexual health risk behaviors in a sample of adult rape survivors (N=102). Women in the 1st cluster (high risk) reported substantial increases from pre- to postrape in their frequency of sexual activity, number of sexual partners, infrequency of condom use, and frequency of using alcohol and/or drugs during sex. The 2nd cluster (moderate risk) reported increases in frequency of sexual activity and number of partners but mitigated that risk with increased condom use. Survivors in the 3rd cluster (low risk) indicated that their sexual health behaviors had become much less risky postrape. An ecological model predicting cluster membership revealed that individual-level and contextual factors predict patterns of risk behaviors. PMID- 14756606 TI - Agonistic interpersonal striving: social-cognitive mechanism of cardiovascular risk in youth? AB - The social competence model (SCM) of cardiovascular disease (CVD) risk identifies combinations of goal-oriented strivings, expressive behaviors, and social skill deficits that contribute to persisting interpersonal difficulties and chronic health-damaging stress in youth. SCM hypotheses were tested on 187 Black and White adolescents who completed the Social Competence Interview (SCI) and later underwent ambulatory blood pressure (ABP) monitoring. Cluster analyses of stress narratives assessed via SCI identified 3 predicted stress profiles: agonistic (interpersonally focused), transcendent (self-development focused), and avoidant. Group comparisons using social, hemodynamic, and ABP data supported the SCM hypothesis that youths who exhibit the agonistic striving profile display diminished social competence, negative social impact, and heightened cardiovascular responding during a stress interview, and elevated ABP during normal social interactions, thus suggesting higher risk of CVD. PMID- 14756607 TI - What is the right thing at the right time? Interactions between stages and processes of change among smokers who make a quit attempt. AB - C. A. Perz, C. C. DiClemente, and J. P. Carbonari (1996) claim support for the transtheoretical model notion that success in smoking cessation involves doing the right thing at the right time: emphasising experiential change processes during the contemplation and preparation stages and shifting to behavioral process activities during action. A key methodological limitation of Perz et al. was their failure to control for stage of change, a measure that has been shown to be predictive of cessation. This study replicates the prospective findings of Perz et al. in a different data set, then controls for stage of change when it is predictive of cessation, and finds that the measures of "appropriate" change process use developed by Perz et al. no longer predict cessation. The authors conclude that stage of change, in particular the distinction between smoking and not smoking, is more important than change process use in predicting cessation outcomes. PMID- 14756608 TI - Emotional distress in nonmetropolitan persons living with HIV disease enrolled in a telephone-delivered, coping improvement group intervention. AB - The study delineated depressive symptoms and modeled emotional distress in persons living with HIV disease in nonmetropolitan areas of 13 U.S. states. Participants (N=329) were enrolled in a randomized clinical trial of a telephone delivered, coping improvement group intervention, and 60% reported moderate or severe levels of depressive symptomatology on the Beck Depression Inventory. Structural equation modeling indicated that participants who experienced more severe HIV symptomatology, received less social support, and engaged in more avoidant coping also experienced more emotional distress (a latent construct comprising depressive symptoms and emotional well-being). Greater HIV-related stigma and rejection by family led to more emotional distress, with social support and avoidant coping mediating almost entirely the effects of the former 2 variables. The model accounted for 72% of the variance in emotional distress in nonmetropolitan persons living with HIV disease. PMID- 14756609 TI - The unconscious cost of good fortune: implicit and explicit self-esteem, positive life events, and health. AB - J. D. Brown and K. L. McGill (1989) found that positive life events were associated with better health only for people high in self-esteem. Among people low in self-esteem, positive life events were associated with poorer health. The authors of this study replicated this finding in a self-report survey of 61 male and 110 female college students. In addition, they showed that implicit self esteem moderated the relation between positive life events and self-reported health in the same fashion as explicit self-esteem did. Whereas people high in implicit self-esteem reported being healthier when they experienced more positive life events, people low in implicit self-esteem reported being healthier when they experienced fewer positive life events. Moreover, the effects of implicit self-esteem were statistically independent of the effects of explicit self esteem. PMID- 14756610 TI - Cognitive trauma therapy for battered women with PTSD (CTT-BW). AB - This article describes a second treatment-outcome study of cognitive trauma therapy for battered women with posttraumatic stress disorder (PTSD; CTT-BW). CTT BW includes trauma history exploration: PTSD education; stress management; exposure to abuse and abuser reminders; self-monitoring of negative self-talk; cognitive therapy for guilt; and modules on self-advocacy, assertiveness, and how to identify perpetrators. One hundred twenty-five ethnically diverse women were randomly assigned to immediate or delayed CTT-BW. PTSD remitted in 87% of women who completed CTT-BW, with large reductions in depression and guilt and substantial increases in self-esteem. White and ethnic minority women benefited equally from CTT-BW. Similar treatment outcomes were obtained by male and female therapists and by therapists with different levels of education and training. Gains were maintained at 3- and 6-month follow-ups. PMID- 14756611 TI - Correlates of levels and patterns of positive life changes following sexual assault. AB - This study builds on previous work suggesting that many survivors report positive life changes soon after a sexual assault and that those who retain those changes over time report the least distress 1 year post-assault (P. Frazier, A. Conlon, & T. Glaser, 2001). The purposes of this study were to assess correlates of early reports of positive life changes and individual trajectories of self-reported positive changes over time among female sexual assault survivors (n = 171) using hierarchical linear modeling. The factors most related to reporting positive life change soon after the assault were social support, approach and religious coping, and perceived control over the recovery process. Increases in these factors also were associated with increases in self-reported positive life changes over time. The relations between social support and positive change also were mediated by coping strategies and control appraisals, particularly perceived control over the recovery process. PMID- 14756612 TI - Mindfulness-based cognitive therapy for depression: replication and exploration of differential relapse prevention effects. AB - Recovered recurrently depressed patients were randomized to treatment as usual (TAU) or TAU plus mindfulness-based cognitive therapy (MBCT). Replicating previous findings, MBCT reduced relapse from 78% to 36% in 55 patients with 3 or more previous episodes; but in 18 patients with only 2 (recent) episodes corresponding figures were 20% and 50%. MBCT was most effective in preventing relapses not preceded by life events. Relapses were more often associated with significant life events in the 2-episode group. This group also reported less childhood adversity and later first depression onset than the 3-or-more-episode group, suggesting that these groups represented distinct populations. MBCT is an effective and efficient way to prevent relapse/recurrence in recovered depressed patients with 3 or more previous episodes. PMID- 14756613 TI - Racial and ethnic differences in depression: the roles of social support and meeting basic needs. AB - The current work examined the depressive symptoms and prevalence of major depression among members of ethnic and racial minorities and White people from a large random sample. Minority group members experienced more depressive symptoms and a marginally higher prevalence of major depression than did White participants. These effects were mediated by participants' problems meeting their basic needs. Specifically, minority group members reported more problems meeting their basic needs, and these problems were associated with an increased risk for depression and depressive symptoms. Minority group members also reported a higher quality of interpersonal functioning than White participants did, which appeared to suppress the relationship between ethnicity and depression. The implications of the findings for treatment are discussed. PMID- 14756614 TI - Eating-disordered behaviors, body fat, and psychopathology in overweight and normal-weight children. AB - This study examined eating-disordered pathology in relation to psychopathology and adiposity in 162 non-treatment-seeking overweight (OW) and normal weight (NW) children, ages 6-13 years. Participants experienced objective or subjective binge eating (S/OBE; loss-of-control eating), objective overeating (OO), or no episodes (NE). OW children experienced significantly higher eating-disordered cognitions and behaviors than NW children and more behavior problems than NW children: 9.3% endorsed S/OBEs, 20.4% reported OOs, and 70.4% reported NEs. OW children reported S/OBEs more frequently than did NW children (p =.01), but similar percentages endorsed OOs. S/OBE children experienced greater eating-disordered cognitions (ps from <.05 to <.01) and had higher body fat (p <.05) than OOs or NEs. OOs are common in childhood, but S/OBEs are more prevalent in OW children and associated with increased adiposity and eating-disordered cognitions. PMID- 14756616 TI - Retrospective and prospective reports of precipitants to relapse in pathological gambling. AB - A prospective design was used to explore the precipitants of relapse in a naturalistic sample of pathological gamblers (N = 101) who had recently quit gambling. Relapse rates were high; only 8% were entirely free of gambling during the 12-month follow-up. Relapses were highly variable but occurred most frequently in the evening, when the person was alone and thinking about finances. Moods prior to the gambling were as likely to be positive as negative. The most frequently reported attributions, particularly for major relapses, were cognitions about winning and feeling the need to make money, unlike substance abuse relapses that tend to be attributed to negative affect. Some gender differences were found, but the precipitants of shorter and longer relapses did not differ. PMID- 14756615 TI - Prospective relations between bulimic pathology, depression, and substance abuse: unpacking comorbidity in adolescent girls. AB - To elucidate the processes that contribute to the comorbidity between bulimic pathology, depression, and substance abuse, the authors tested the temporal relations between these disturbances with prospective data from adolescent girls (N = 496). Multivariate analyses indicated that depressive symptoms predicted onset of bulimic pathology but not of substance abuse, bulimic symptoms predicted onset of depression but not of substance abuse, and substance abuse symptoms predicted onset of depression but not of bulimic pathology. Results suggest that the comorbidity arises because certain disorders are risk factors for the other disorders. Findings also provide support for select etiologic theories and further establish the clinical significance of these conditions by showing that they increase risk for onset of other psychiatric disturbances. PMID- 14756617 TI - Long-term influence of duration and frequency of participation in alcoholics anonymous on individuals with alcohol use disorders. AB - This study examined the influence of the duration and frequency of a baseline episode of participation in Alcoholics Anonymous (AA) among 473 individuals with alcohol use disorders on 1-year and 8-year outcomes and the effect of additional participation and delayed participation on outcomes. Compared with individuals who did not participate, individuals who affiliated with AA relatively quickly, and who participated longer, had better 1-year and 8-year alcohol-related outcomes. Individuals who continued to participate, and those who continued longer, had better alcohol-related outcomes than did individuals who discontinued participation, but individuals who delayed participation in AA had no better outcomes than those who never participated. In general, the frequency of participation was independently associated only with a higher likelihood of abstinence. PMID- 14756618 TI - En las manos de Dios [in God's hands]: Religious and other forms of coping among Latinos with arthritis. AB - This study tested a theoretical model concerning religious, passive, and active coping; pain; and psychological adjustment among a sample of 200 Latinos with arthritis. Respondents reported using high levels of religious coping. A path analysis indicated that religious coping was correlated with active but not with passive coping. Religious coping was directly related to psychological well being. Passive coping was associated with greater pain and worse adjustment. The effects of active coping on pain, depression, and psychological well-being were entirely indirect, mediated by acceptance of illness and self-efficacy. These findings warrant more research on the mechanisms that mediate the relationship between coping and health. This study contributes to a growing literature on religious coping among people with chronic illness, as well as contributing to a historically under-studied ethnic group. PMID- 14756619 TI - Childhood peer rejection and aggression as predictors of adolescent girls' externalizing and health risk behaviors: a 6-year longitudinal study. AB - This 6-year longitudinal study examined girls' peer-nominated social preference and aggression in childhood as predictors of self- and parent-reported externalizing symptoms, substance use (i.e. cigarette, alcohol, and marijuana use), and sexual risk behavior in adolescence. Participants were 148 girls from diverse ethnic backgrounds, who were initially assessed in Grades 4-6 and again in Grades 10-12. Results supported a moderator model, indicating that social preference changed the nature of the association between childhood aggression and adolescent outcomes. When accompanied by peer rejection, aggressive behavior was moderately stable over time and significantly associated with adolescent girls' substance use and sexual risk behavior. However, under conditions of peer acceptance, no significant association between childhood aggression and adolescent outcomes emerged. PMID- 14756620 TI - Internet-based treatment for insomnia: a controlled evaluation. AB - This study investigated the effects of an Internet-based intervention for insomnia. Participants who met criteria for insomnia (N = 109) were randomly assigned to either a cognitive-behavioral self-help treatment or a waiting list control condition. The 5-week intervention mainly consisted of sleep restriction, stimulus control, and cognitive restructuring. Sleep diary data were collected for 2 weeks at baseline and at posttreatment. The dropout rate was 24% (n = 28). Results showed statistically significant improvements in the treatment group on many outcome measures, including total sleep time, total wake time in bed, and sleep efficiency. However, improvements were also found in the control group. Overall, between-groups effect sizes were low, with the exception of the Beliefs and Attitudes About Sleep Scale (Cohen's d =.81). PMID- 14756621 TI - Preliminary reliability and validity of the Clinician-Administered PTSD Scale for schizophrenia. AB - This study provides preliminary psychometric support for a version of the Clinician-Administered Posttraumatic Stress Disorder (PTSD) Scale (CAPS; D. D. Blake et al., 1990) adapted for use with patients with schizophrenia (CAPS-S; J. S. Gearon. S. Thomas-Lohrman, & A. S. Bellack, 2001). Nineteen women with schizophrenia and co-occurring illicit drug use disorders were administered the CAPS-S, the Structured Clinical Interview for DSM-IV diagnoses (SCID). and scales measuring trauma-related psychopathology. The results indicate that the CAPS-S can distinguish between those with and without PTSD and that the symptom clusters measure unified constructs. Interrater and test-retest reliability were high for PTSD diagnosis and symptom clusters. Solid convergent validity was demonstrated between the CAPS-S and SCID-based PTSD diagnoses and the Impact of Event Scale. There is also preliminary evidence of discriminant validity. These results support the use of the CAPS-S in women with schizophrenia. PMID- 14756622 TI - Diagnostic efficiency of DSM-IV criteria for borderline personality disorder: an evaluation in Hispanic men and women with substance use disorders. AB - This study examined diagnostic efficiency of Diagnostic and Statistical Manual of Mental Disorders, fourth edition (DSM-IV), criteria for borderline personality disorder (BPD). One hundred thirty monolingual Hispanic adults (90 men, 40 women) at an outpatient psychiatric and substance abuse clinic were assessed with the Spanish-Language Version of the Diagnostic Interview for DSM-IV Personality Disorders (C. M. Grilo, L. M. Anez, & T. H. McGlashan, 2003). The BPD diagnosis was determined by the best-estimate method. Diagnostic efficiency indices were calculated for all BPD criteria, for the entire study group, and separately by gender. Overall, the best exclusion criterion was affective instability, whereas suicidality or self-injury was the best inclusion criterion and the best predictor overall. These findings did not differ by gender, are similar to those reported elsewhere in the literature, and have implications for the refinement of diagnostic systems. PMID- 14756623 TI - The tumor cell and telomerase. AB - Imbalanced activity of the mechanism that controls cell division is a prerequisite for malignant transformation of a normal cell. The present review considers this multi-step mechanism, which is usually called the G1-S checkpoint. Besides, tumor cells are characterized by the presence of telomerase, an enzyme responsible for restoration of chromosome ends after replication and thus providing for unlimited cell division. The main point of the present article is to find out whether the activation of telomerase is controlled by the G1-S checkpoint or does not depend on it. The principal components of the G1-S checkpoint, such as cyclin-dependent kinases, retinoblastoma and E2F proteins, control the activity of telomerase. In their turn, they accumulate and transmit signals from various sources inside and outside the cell. Thus, various changes in tumor cells can activate telomerase through the G1-S checkpoint. Such are the suggested effects on telomerase of Myc, p53, Waf1, protein kinases B and C, Wnt5A, TGFbeta, WT1, and estrogens. However, Myc, p53, WT1, estrogens, protein kinases B and C, and TGFbeta can also directly influence telomerase independently of the G1-S checkpoint mechanism. Moreover, in 30% of human tumors the gene of the key subunit of telomerase (hTERT) is amplified, possibly due to chromosomal rearrangements unassociated with the activity of the G1-S checkpoint. Thus, telomerase seems to be activated not by a single agent but due to combined action of various factors, both with involvement of the G1-S checkpoint mechanism and independently of it. PMID- 14756624 TI - Integrins: structure and signaling. AB - Integrins are cell surface transmembrane glycoproteins that function as adhesion receptors in cell-extracellular matrix interactions and link the matrix proteins to the cytoskeleton. The family of human integrins comprises 24 members, each of which is a heterodimer consisting of 1 of 18 alpha- and 1 of 8 beta-subunits. Integrins play an important role in the cytoskeleton organization and in transduction of intracellular signals, regulating various processes such as proliferation, differentiation, apoptosis, and cell migration. This review summarizes current views on the structure of integrins, integrin associated proteins, and biochemical mechanisms underlying their signaling functions. PMID- 14756625 TI - Special features of the DNA-hydrolyzing activity of the antibodies in systemic lupus erythematosus. AB - Two types of IgG anti-DNA antibodies exhibiting DNA-hydrolyzing activity have been isolated from blood serum of patients with systemic lupus erythematosus. This DNase activity of antibodies differs from serum DNases by the non-processive mode, temperature resistance, pH optimum, and the rate of DNA hydrolysis. It is suggested that the anti-DNA antibody molecule possessing DNase activity contains two sites: one site determines specificity of antibody-DNA interaction, whereas the other is responsible for manifestation of the catalytic activity. PMID- 14756626 TI - Thermostable DNA polymerase from Thermus thermophilus B35: preparation and study of a modified form of the enzyme with high affinity to ddNTP. AB - The hybrid protein consisting of Tte DNA polymerase fragment and mutant Taq DNA polymerase (F667Y) fragment in the ratio 20 : 1 was constructed. Affinity of the modified enzyme (substitutions F669Y, V667I, and S692Q) to ddNTP was two orders higher than that of the wild type enzyme. The modified enzyme was used for sequencing DNA fragment with total deoxyguanosine and deoxycytidine content of 68%. In the polymerase chain reaction, the modified enzyme exhibits properties typical of the wild type Tte DNA polymerase. PMID- 14756627 TI - Recombinant endo-beta-1,4-xylanase from Penicillium canescens. AB - Recombinant endo-beta-1,4-xylanase (Xyl-31rec, 31 kD, pI 8.2-9.3, the tenth family of glycosyl hydrolases) was isolated from the culture liquid of Penicillium canescens (strain with the amplified homologous xylanase gene) by chromatofocusing on Mono P and hydrophobic chromatography on phenyl-Superose. It is shown that the biochemical and kinetic parameters, substrate specificity, stability, and other properties of the recombinant and native enzymes are almost the same. It was found that Xyl-31rec can be used for biobleaching of cellulose, the recombinant P. canescens strains providing a high yield of extracellular Xyl 31rec (up to 800-900 U/ml of culture liquid) and not secreting cellulases. PMID- 14756628 TI - Proton pumping in growing part of maize root: its correlation with 14-3-3 protein content and changes in response to osmotic stress. AB - The spatial pattern of mitotic activity, cell elongation, rate of H+ fluxes, and 14-3-3 protein content were determined in Zea mays roots. We found that the regions along the apical part of the growing root conversely differ in their proton pumping activity. Higher rate of H+ efflux coincides with higher growth rate and correlates with increased 14-3-3 protein content in membrane preparations. The segment consisting of the root cap and the apical part of the meristem exerts net inward proton pumping, which can be inverted under fusicoccin treatment or osmotic stress. In the latter case, this inversion is accompanied by accumulation of 14-3-3 protein in plasma membranes. The results obtained highlight 14-3-3 protein as an obvious candidate for the fine regulation of plasma membrane H+-ATPase in root apex. PMID- 14756629 TI - Acid phosphatase complex from the freshwater snail Viviparus viviparus L. under standard conditions and intoxication by cadmium ions. AB - Acid phosphatases differing in both subcellular localization and substrate specificity were isolated for the first time from the liver of the freshwater snail Viviparus viviparus L. by preparative isoelectrofocusing. One of five characterized phosphatases is highly specific to ADP and the others can hydrolyze (at variable rate) a series of natural substrates. A scheme is proposed for the involvement of the studied phosphatases in carbohydrate metabolism. We have also studied some peculiarities of the effect of Cd2+ in vitro and in vivo on the activities of individual components of the acid phosphatase complex and corresponding changes in metabolism of the freshwater snail as a new test-object allowing the estimation of toxicity in water. PMID- 14756630 TI - Oligonucleosomal DNA fragmentation in MCF-7 cells undergoing palmitate-induced apoptosis. AB - Oligonucleosomal fragmentation of nuclear DNA is the late-stage apoptosis hallmark. In apoptotic mammalian cells the fragmentation is catalyzed by DFF40/CAD DNase primarily activated by caspase 3 through the site-specific proteolytic cleavage of DFF45/ICAD. A deletion in the casp3 gene of human breast adenocarcinoma MCF-7 results in lack of procaspase 3 in these cells. The absence of caspase 3 in MCF-7 leads to disability to activate oligonucleosomal DNA fragmentation in TNF-alpha induced cell death. In this study, sodium palmitate was used as an apoptotic stimulus for MCF-7. It has been shown that palmitate but not TNF-alpha induces both apoptotic changes in nuclei and oligonucleosomal DNA fragmentation in casp3-mutated MCF-7. Activation and accumulation of 40-50 kD DFF40-like DNases in nuclei of palmitate-treated apoptotic MCF-7 were detected by SDS-DNA-PAGE assay. Microsomal fraction of apoptotic MCF-7 does not contain any detectable DNases, but activates 40-50 kD nucleases when incubated with human placental chromatin. Furthermore, microsomes of apoptotic MCF-7 induce oligonucleosomal fragmentation of chromatin in a cell-free system. Both the activation of DNases and chromatin fragmentation are suppressed in the presence of the caspase 3/7 inhibitor Ac-DEVD-CHO. Microsome-associated caspase 7 is suggested to play an essential role in the induction of oligonucleosomal DNA fragmentation in casp3-deficient MCF-7 cells. PMID- 14756631 TI - Nucleoside diphosphate kinase is a possible component of the ethylene signal transduction pathway. AB - In etiolated seedlings of Pisum sativum and leaves of Arabidopsis thaliana, in vivo ethylene treatment resulted in an increase in in vitro phosphorylation of 17 kD (P. sativum) or 16 and 17 kD (A. thaliana) polypeptides. These polypeptides were identified as nucleoside diphosphate kinase (NDPK) based on both biochemical properties and interaction with antibodies against NDPK from P. sativum. Using the receptor-directed antagonist of ethylene action 2,5-norbornadiene and the ethylene-insensitive mutants of A. thaliana etr1-1 and eti5, ethylene specificity and receptor dependence of NDPK phosphorylation have been demonstrated. In pea epicotyls, ethylene treatment also led to increase in nucleoside transferase activity unlike in A. thaliana leaves. The increases in nucleoside transferase activity and NDPK phosphorylation were very rapid and transient. The results suggest a role for NDPK as a possible component of the ethylene signal transduction chain. PMID- 14756633 TI - Structure of silenan, a pectic polysaccharide from Campion Silene vulgaris (Moench) Garcke. AB - A pectic polysaccharide named silenan, [alpha]D20 +148.6 degrees (c 0.1; H2O), was isolated earlier from the aerial part of campion, Silene vulgaris (Moench) Garcke. Silenan has been shown to contain homogalacturonan segments as "smooth regions" and rhamnogalacturonan fragments as "hairy regions". The present study reveals a generalization of structural features of silenan. Silenan was subjected to enzymic digestion with pectinase, to Smith degradation, and to lithium degradation to determine the conforming poly- and oligosaccharide fragments of "hairy regions" of silenan. The NMR-spectral data and mass-spectrometry confirmed that the core of the ramified region of silenan consisted of residues of alpha rhamnopyranose 2-O-glycosylated with the residues of alpha-1,4-D-galactopyranosyl uronic acid. The part of the alpha-rhamnopyranose residues of the backbone are branched at O-4. On the basis of the data, the hairy regions of silenan proved to contain mainly linear chains of beta-1,3-, beta-1,4-, and beta-1,6-galactopyranan and alpha-1,5-arabinofuranan. The side chains of the ramified region were shown to have branching points represented 2,3-, 3,6-, 4,6-di-O-substituted beta galactopyranose residues. PMID- 14756632 TI - Mechanism of superoxide anion generation in intact mitochondria in the presence of lucigenin and cyanide. AB - In the presence of cyanide and various respiratory substrates (succinate or pyruvate + malate) addition of high concentrations of lucigenin (400 microM; Luc2+) to rat liver mitochondria can induce a short-term flash of high amplitude lucigenin-dependent chemiluminescence (LDCL). Under conditions of cytochrome oxidase inhibition by cyanide the lucigenin-induced cyanide-resistant respiration (with succinate as substrate) was not inhibited by uncouplers (FCCP) and oligomycin. Increase in transmembrane potential (Deltaphi) value by stimulating F0F1-ATPase functioning (induced by addition of MgATP to the incubation medium) caused potent stimulation of the rate of cyanide-resistant respiration. At high Deltaphi values (in the presence of MgATP) cyanide resistant respiration of mitochondria in the presence of succinate or malate with pyruvate was insensitive to tenoyltrifluoroacetone (TTFA) or rotenone, respectively. However, in both cases respiration was effectively inhibited by myxothiazol or antimycin A. Mechanisms responsible for induction of LDCL and cyanide resistant mitochondrial respiration differ. In contrast to cyanide-resistant respiration, generation of LDCL signal, that was suppressed only by combined addition of Complex III inhibitors, antimycin A and myxothiazol, is a strictly potential-dependent process. It is observed only under conditions of high Deltaphi value generated by F0F1-ATPase functioning. The data suggest lucigenin-induced intensive generation of superoxide anion in mitochondria. Based on results of inhibitor analysis of cyanide-resistant respiration and LDCL, a two-stage mechanism of autooxidizable lucigenin cation-radical (Luc*+) formation in the respiratory chain is proposed. The first stage involves two-electron Luc2+ reduction by Complexes I and II. The second stage includes one-electron oxidation of reduced lucigenin (Luc(2e)). Reactions of Luc(2e) oxidation involve coenzyme Q-binding sites of Complex III. This results in formation of autooxidizable Luc*+ and superoxide anion generation. A new scheme for lucigenin-dependent electron pathways is proposed. It includes formation of fully reduced form of lucigenin and two-electron transferring shunts of the respiratory chain. Lucigenin-induced activation of superoxide anion formation in mitochondria is accompanied by increase in ion permeability of the inner mitochondrial membrane. PMID- 14756634 TI - Mechanism of perfluoroalkyl halide toxicity: catalysis of perfluoroalkylation by reduced forms of cobalamin (vitamin B12). AB - Perfluoroalkyl halides (PFHs) are synthetic products widely used in various fields. Perfluorooctyl bromide (PFB) is used in medicine as a component of blood substitutes and for artificial lung ventilation. In both cases, it is considered a completely inert compound acting as a solvent for oxygen. However, there are many reports of PFH-induced intoxication, including lethal cases. Mechanisms underlying toxic effects of this compound remain unknown. In this study, we demonstrate that the reduced form of cobalamin (vitamin B12) typical for B12 dependent enzymes can catalyze the reactions of perfluoroalkylation, aromatic substitution, or addition by double bonds. Synthesis of perfluoro derivatives from PFHs during catalysis by cob(I)alamin-like super nucleophiles is a new possible mechanism responsible for in vivo formation of highly toxic compounds from "chemically inert" substances widely used in medicine. Catalytic perfluoroalkylation might possibly contribute to nitric oxide depletion and modulation of activity of guanylate cyclase, cytochromes, NO-synthases, and other heme-containing proteins. PMID- 14756635 TI - Continuous positive airway pressure decreases myocardial oxygen consumption in heart failure. AB - The aim of the present study was to investigate the effects of CPAP (continuous positive airway pressure) support on myocardial energetics in patients with CHF (congestive heart failure). CPAP has been shown to decrease left ventricular afterload and to produce favourable short- and long-term haemodynamic and neurohormonal benefits in CHF patients. The mechanisms responsible for these actions are not completely understood. We measured the haemodynamic and myocardial metabolic response to the acute (10 min) application of CPAP in CHF patients. Myocardial VO(2) (O(2) consumption) and VCO(2) (CO(2) production) were measured by simultaneous arterial and coronary sinus blood sampling. The application of CPAP resulted in a significant decrease in left ventricular stroke work (97+/-12 to 83+/-9 g.m; P <0.05) and myocardial VO(2) (0.32+/-0.03 to 0.25+/ 0.01 ml of O(2)/beat; P <0.05). Myocardial mechanical efficiency, however, was unchanged. CPAP application decreases myocardial work and VO(2). This effect on myocardial energetics could account for some of the favourable effects of CPAP in CHF patients. PMID- 14756637 TI - Noninvasive lower eyelid blepharoplasty: a new technique using nonablative radiofrequency on periorbital skin. AB - BACKGROUND: Laxity and rhytids of the lower eyelids are common cosmetic concerns. Historically, correction has either been surgical through either transcutaneous or transconjunctival blepharoplasty or ablative through laser resurfacing or chemical peeling. Therapeutic options usually require significant postoperative healing and have the potential risk of scarring ectropion or pigmentary loss. OBJECTIVE: To report the use of a new technique that uses nonablative radiofrequency (NARF) to tighten noninvasively and nonsurgically the flaccid skin of the lower eyelids by treating the periorbital area to produce cosmetic improvement. METHODS: Nine patients with skin flaccidity of the lower eyelids had a single treatment session with NARF in a small area of skin in the periorbital region, specifically the zygomatic and/or temporal areas. All patients were treated with topical anesthesia only. The treatment lasted approximately 10 minutes. No postoperative care was required. RESULTS: All of the nine patients in the study achieved cosmetic improvement of the eyelids ostensibly through skin contraction. All patients were able to return to their normal routines immediately. Although the results were gradual, patient satisfaction was remarkable. No complications were seen in this study. CONCLUSION: This new procedure using NARF was successful in providing a safe, noninvasive, cosmetic improvement in these patients with excessive skin laxity of the lower eyelids. Postoperative morbidity, including down time and complications, was not seen. PMID- 14756638 TI - Combined excimer laser and topical tacrolimus for the treatment of vitiligo: a pilot study. AB - BACKGROUND: Vitiligo is an acquired skin disorder that is characterized by well defined, often symmetric white patches. Although current therapeutic modalities are directed toward increasing melanocyte melanin production, few treatment modalities address the immunologic nature of the disease. OBJECTIVE: To determine whether excimer laser, a known therapeutic modality, in combination with tacrolimus, a topical immunomodulator, accelerate response time and/or improve the degree of response in patients with this disorder. METHODS: Eight subjects diagnosed with vitiligo were recruited to participate in this institutional review board-approved double-blind, placebo-controlled study. Twenty-four symmetric vitiliginous patches (elbows, knees) from eight subjects received excimer laser treatment three times per week for 24 treatments or 10 weeks. Additionally, topical tacrolimus 0.1% ointment (Protopic) and placebo (Aquaphor) were applied to randomized patches (left or right) twice daily throughout the length of the trial. Vitiliginous patches were monitored with photographs at baseline, every 2 weeks, and 6 months after treatment. Biopsies were performed on subjects with significant results. RESULTS: Twenty vitiliginous patches from six subjects qualified for evaluation. Fifty percent of patches treated with combination excimer laser and tacrolimus achieved a successful response (75% repigmentation) compared with 20% for the placebo group. Subjects who responded successfully repigmented faster (19%) with combination therapy compared with excimer laser alone. Additionally, three subjects experienced transient hyperpigmentation in lesions treated with combination therapy. CONCLUSION: Combining topical immunomodulators with known phototherapeutic modalities may represent a key advancement in the treatment of disease. PMID- 14756639 TI - Workforce characteristics of mohs surgery fellows. AB - BACKGROUND: Anecdotal evidence from program directors and Mohs surgeons suggests that Mohs fellowships are becoming increasingly popular and competitive among dermatology trainees. OBJECTIVE: To assess the characteristics and investigate the motivating factors of those pursuing Mohs fellowships. METHODS: Anonymous surveys were distributed to recent dermatology residency graduates taking a board exam review course in years 1999-2002. RESULTS: In 2002, 2001, and 1999, the percentages of recently trained dermatologists pursuing Mohs fellowships were 9.4%, 8.5%, and 8.8%, respectively. There were no significant differences between Mohs fellows and the rest of the recently graduated dermatologists in terms of debt levels, marital status, parenting status, and spousal employment status. The Mohs fellows were slightly more likely to be male than their non-Mohs counterparts. The factor considered the most important by both groups when choosing a job was location. CONCLUSIONS: Further research is needed to discover potential factors that may be playing a role in the increased popularity of Mohs surgery. The number of Mohs surgeons is increasing and is likely to expand over time. It remains to be seen what effect the growth will have on the supply of Mohs surgery and whether it will outpace the increased demand for services. PMID- 14756640 TI - Light therapy in the treatment of acne vulgaris. AB - BACKGROUND: Over the past decade, lasers and light-based systems have become a common modality to treat a wide variety of skin-related conditions, including acne vulgaris. In spite of the various oral and topical treatments available for the treatment of acne, many patients fail to respond adequately or may develop side effects. Therefore, there is a growing demand by patients for a fast, safe, and side-effect-free novel therapy. OBJECTIVES: To address the role of light therapy in the armamentarium of treatments for acne vulgaris, to discuss photobiology aspects and biomedical optics, to review current technologies of laser/light-based devices, to review the clinical experience and results, and to outline clinical guidelines and treatment considerations. RESULTS: Clinical trials show that 85% of the patients demonstrate a significant quantitative reduction in at least 50% of the lesions after four biweekly treatments. In approximately 20% of the cases, acne eradication may reach 90%. At 3 months after the last treatment, clearance is approximately 70% to 80%. The nonrespondent rate is 15% to 20%. CONCLUSIONS: Laser and light-based therapy is a safe and effective modality for the treatment of mild to moderate inflammatory acne vulgaris. Amelioration of acne by light therapy, although comparable to the effects of oral antibiotics, offers faster resolution and fewer side effects and leads to patient satisfaction. PMID- 14756641 TI - Treatment of inflammatory facial acne vulgaris with the 1450-nm diode laser: a pilot study. AB - BACKGROUND: The 1450-nm diode laser has been found to damage sebaceous glands selectively and to be effective for the treatment of inflammatory acne on the back. OBJECTIVE: To evaluate the efficacy and safety of the 1450-nm diode laser in the treatment of inflammatory facial acne vulgaris. METHODS: Nineteen patients with inflammatory facial acne were treated with the 1450-nm diode laser at 4- to 6-week intervals. There was no control group. Clinical photographs and lesion counts were obtained at baseline and after each treatment. Subjective evaluation of response to treatment and pain was assessed using a questionnaire. RESULTS: All patients had a reduction in acne lesions. Lesion counts decreased 37% after one treatment (p<0.01), 58% after two treatments (p<0.01), and 83% after three treatments (p<0.01). Treatment-related pain was well tolerated, and adverse effects were limited to transient erythema and edema at treatment sites. CONCLUSION: This is the first published report documenting the safety and efficacy of laser treatment for inflammatory facial acne. In our study, clinical improvement was seen in all patients and was generally dramatic, even in those refractory to previous treatment with oral isotretinoin. Topical anesthetics should be used to minimize pain associated with treatment. PMID- 14756642 TI - Comparison of a 1450-nm diode laser and a 1320-nm Nd:YAG laser in the treatment of atrophic facial scars: a prospective clinical and histologic study. AB - BACKGROUND: Atrophic scar revision techniques, although numerous, have been hampered by inadequate clinical responses and prolonged postoperative recovery periods. Nonablative laser treatment has been shown to effect significant dermal collagen remodeling with minimal posttreatment sequelae. Although many studies have been published regarding the effectiveness of these nonablative lasers on rhytides, there are limited data demonstrating their specific effects on atrophic scars. OBJECTIVE: To evaluate and compare the efficacy and safety of long-pulsed 1320-nm Nd:YAG and 1450-nm diode lasers in the treatment of atrophic facial scarring. METHODS: A series of 20 patients (skin phototypes I-V) with mild to moderate atrophic facial acne scars randomly received three successive monthly treatments with a long-pulsed 1320-nm Nd:YAG laser on one facial half and a long pulsed 1450-nm diode laser on the contralateral facial half. Patients were evaluated using digital photography and three-dimensional in vivo microtopography measurements at each treatment visit and at 1, 3, 6, and 12 months postoperatively. Histologic evaluations of cutaneous biopsies obtained before treatment, immediately after the first treatment, and at 1, 3, 6, and 12 months after the third treatment were performed. Clinical assessment scores were determined at each treatment session and follow-up visit. Patient satisfaction surveys were obtained at the end of the study. RESULTS: Mild to moderate clinical improvement was observed after the series of three treatments in the majority of patients studied. Patient satisfaction scores and in vivo microtopography measurements paralleled the photographic and histopathologic changes seen. Side effects of treatment were limited to mild transient erythema, edema, and hyperpigmentation. No scarring or adverse textural changes resulted from the use of either laser system. CONCLUSIONS: Nonablative long-pulsed 1320-nm Nd:YAG and 1450-nm diode lasers each offer clinical improvement for patients with atrophic scarring without significant side effects or complications. The 1450-nm diode laser showed greater clinical scar response at the parameters studied. The use of nonablative laser systems is a good treatment alternative for patients with atrophic scarring who are unable or unwilling to endure the prolonged postoperative recovery process associated with ablative laser skin resurfacing procedures. PMID- 14756643 TI - Evaluating the efficacy of treatment of resistant port-wine stains with variable pulse 595-nm pulsed dye and 532-nm Nd:YAG lasers. AB - BACKGROUND: Some port wine stains (PWSs), despite multiple treatments with the 585-nm 0.45-ms pulsed dye laser (PDL), fail to improve substantially. OBJECTIVE: To determine the efficacy and tolerability of variable pulse width 595-nm PDL and 532-nm Nd:YAG laser in the treatment of resistant PWS. METHODS: Twenty-two patients whose PWS failed to achieve more than 75% lightening after more than 15 treatments with the 585-nm 0.45-ms PDL were recruited. A homogenous patch of PWS was divided into five areas. Area 1 (control area) was treated with 585-nm, 0.45 ms PDL (fluence 7.5 J/cm2). Areas 2 and 3 were treated with 595-nm PDL at fluence 15 J/cm2 (with cryogen spray cooling) and pulse durations of 1.5 and 10 ms, respectively. Areas 4 and 5 were treated with a 532-nm Nd:YAG laser at 2 ms, 7 J/cm2 and 10 ms, 16 J/cm2, respectively (with a contact cooling tip). The response was assessed by photographic evaluation. RESULTS: Three patients had further lightening in area 2, and two patients had further lightening in area 3. Each of three patients had further lightening in areas 4 and 5, respectively. One patient had further lightening in the control area. CONCLUSION: In individual patients, it may be effective to treat resistant PWS with the variable-pulse width 595-nm PDL and the 532-nm Nd:YAG laser. PMID- 14756644 TI - Does pulse stacking improve the results of treatment with variable-pulse pulsed dye lasers? AB - BACKGROUND: It has been suggested that multiple stacked pulses of lower fluence may have a similar effect on targets as a single pulse of higher fluence. When treating vascular lesions, increasing the fluence beyond a certain point will increase the risk of purpura given a constant pulse duration. Stacking pulses of lower fluence may have the advantage of heating vessels to a critical temperature without creating purpura. OBJECTIVE: To determine whether stacking low-fluence pulses of a variable-pulse pulsed-dye laser would improve clinical results without significantly increasing side and adverse effects. METHODS: Twenty-five patients between the ages of 18 and 65 years with facial telangiectasia and skin types I-IV were enrolled in the study. For each subject, the cheek or nasal ala areas on either side of the facial midline with similar telangiectasia density ratings were randomized to single pulse and multiple stacked pulse groups. One side of the cheek or nasal ala was treated with single nonoverlapping pulses with the Candela Vbeam 595-nm pulsed-dye laser. The opposite side of the cheek or nose was treated with the same parameters but with three or four pulses stacked on top of each other at a 1.5-Hz repetition rate. Patients were asked to rate the pain of the procedure on each side on a 0 to 3 scale. Investigators rated the erythema and edema after the procedure as well as vessel clearing and overall telangiectasia density scale at 1 and 6 weeks after the procedure. RESULTS: Twenty-three patients completed the study. The mean pain rating was 1.58 for the pulse stacked side and 1.38 for the single-pass side. The mean erythema score after the procedure was 1.17 for the pulsed stacked side and 1.09 for the single pulsed side. The mean vessel clearing 1 week after the treatment was 74.3% for the pulse stacked side and 58.5% for the single pulsed side. The mean vessel clearing 6 weeks after the treatment was 87.6% for the pulse stacked side and 67.4% for the single pulsed side. The mean telangiectasia density scale score before treatment was 2.67 for the pulse stacked side and 2.59 for the single pulsed side. At 1 week after treatment, the mean telangiectasia density scale score was 1.06 for the pulsed stacked side and 1.5 for the single pulsed side. At 6 weeks after treatment, the mean telangiectasia density scale score was 0.72 for the pulsed stacked side and 1.30 for the single pulsed side. No patients experienced purpura in either group, and there were no cases of hyperpigmentation, hypopigmentation, or scar formation. One patient experienced significant edema on the side of the cheeks treated with pulse stacking. CONCLUSIONS: Treating superficial facial telangiectasia with a pulse stacking technique may improve clinical results without significantly increasing adverse effects. PMID- 14756645 TI - Hair reduction using intense pulsed light source. AB - BACKGROUND: Long-term hair removal in hirsute women remains a challenging issue. Various laser and laser-like devices are currently in use for hair removal, but little is known about the permanence of their results. This study deals with the permanence of hair removal using the intense pulsed light source (IPLS). OBJECTIVE: To test the effectiveness in long-term hair reduction. METHODS: Seventy female hirsute patients were selected in the Department of Laser Therapy at the Medical Center, Maastricht, the Netherlands. The average age of the mostly dark-haired patients of various skin types (Fitzpatrick I to V) was 41 years. They were subjected to a mean of 8 treatments (range of 2 to 23) followed for a mean period of 27.3 months. RESULTS: Using the IPLS, 87% hair removal was achieved, whereby the number of treatments correlated with the amount of hair lost. No correlation was found between hair removal and patient-related or technical data. Minimal side effects occurred in 10% of the patients. CONCLUSION: The IPLS system with its broad range of technical variables is effective in achieving long-term hair removal. PMID- 14756646 TI - Nonocclusion and early reopening of the great saphenous vein after endovenous laser treatment is fluence dependent. AB - BACKGROUND: Parameters influencing failure and recanalization rates of endovenous laser treatment (ELT) of the great saphenous vein (GSV) are still to be determined. OBJECTIVE: To evaluate treatment-related parameters of ELT with respect to early failure of occlusion or recanalization of GSVs. METHODS: A series of 77 consecutive patients received ELT of 106 GSVs with continuous pullback of the laser fiber. Duplex examination was performed at 1 day, 4 weeks, and 3 months after the procedure. Clinical patient and vessel characteristics as well as technical parameters of the ELT procedure were evaluated via multiple logistic regression analysis. RESULTS: A median vein length of 60 cm (range of 18 to 90) was treated with a median pullback velocity of 0.6 cm/sec (range of 0.4 to 1.3), resulting in a median energy delivery of 23.4 J/cm (range of 11.8 to 35.5) and a median laser fluence of 11.8 J/cm2 (range of 2.8 to 37.3). At day 1 after ELT, 6 GSVs (6%) were not occluded. At 1 and 3 months after ELT, 9 GSVs (9%) and 11 GSVs (10%), respectively, were found open by color duplex examination. Risk factors for nonocclusion 3 months after ELT, by univariate analysis, were laser fluence, laser energy per centimeter of vein length, diameter of the vein before treatment, and distance of the thrombus to the sapheno-femoral junction at day 1 after treatment. Finally, multiple regression analysis selected laser fluence (p=0.004, odds ratio=0.40 J/cm2) as the relevant risk factor for ELT failure or recanalization. CONCLUSION: ELT failure seems to be related to the administration of low laser fluences. PMID- 14756647 TI - Trichloroacetic acid peeling versus dermabrasion: a histometric, immunohistochemical, and ultrastructural comparison. AB - BACKGROUND: Trichloroacetic acid (TCA) chemical peel and dermabrasion are beneficial methods for treatment of photoaged skin. OBJECTIVE: In this study, we evaluated the changes induced by these therapies on various structures of facial skin of nine dark-skinned patients (Fitzpatrick types IV-V; TCA, five patients; dermabrasion, four patients) demonstrating different degrees of photodamage. METHODS: Routine histopathology coupled with histometric computer-assisted image analysis was used to assess epidermal changes. Alcian blue stain was used to evaluate changes in glycosaminoglycans. Immunoperoxidase techniques with antibodies against types I and III collagen and elastin were used to evaluate quantitatively changes in collagen and elastic fibers, and their ultrastructure was examined by transmission electron microscopy. RESULTS: Similar histologic, immunohistochemical, as well as ultrastructural changes were observed in the two groups, including epidermal and dermal rejuvenation with new collagen deposition and normalization of the elastic tissue. However, these changes were more prominent in patients treated with dermabrasion than those treated with TCA. CONCLUSION: The results of this study suggest that beneficial effects of such modalities on facial skin were accomplished primarily by increasing the amounts of collagen I and collagen III and improving the morphologic appearance of collagen and elastic fibers. PMID- 14756649 TI - Deep plane fixation in integumental surgery. AB - BACKGROUND: Standard wound closure techniques are prone to complication in the presence of tension. OBJECTIVE: To show that deep plane fixation (DPF), a surgical modality based on limited undermining and strategic placement of DPF sutures, affects tension-reduced closure in wounds that would otherwise require skin grafts/flaps or tissue expansion. METHODS: The study is based on an analysis of over 2000 scalp operations. RESULTS: Two groupings of identical sagittal scalp reductions were done. DPF was used in one and not the other. In the series with DPF, reduced tension closure was consistently possible, as was significantly increased excision, relative to the series without DPF. CONCLUSIONS: DPF narrows the wound base and channels tension forces from superficial to deep and/or nonundermined tissues (where they harmlessly dissipate). This allows relatively increased tissue excision and tension-reduced closure. PMID- 14756648 TI - The polyhydroxy acid gluconolactone protects against ultraviolet radiation in an in vitro model of cutaneous photoaging. AB - BACKGROUND: Ultraviolet (UV) radiation damages skin through a variety of mechanisms, including the generation of free radicals. Gluconolactone is a polyhydroxy acid (PHA) that is capable of chelating metals and may also function by scavenging free radicals, thereby protecting skin from some of the damaging effects of UV radiation. OBJECTIVE: This study measured the ability of gluconolactone to protect against UV radiation-induced damage. METHODS: The ability of gluconolactone to prevent UV radiation-induced elastin promoter activation was determined in vitro using a transgenic model of cutaneous photoaging. Gluconolactone was also evaluated to determine its ability to promote the formation of sunburn cells in human skin after exposure to UV radiation. RESULTS: Gluconolactone provided up to 50% protection against UV radiation, as measured in our in vitro system, and did not significantly increase sunburn cells in human skin. CONCLUSIONS: These results demonstrate the ability of the PHA gluconolactone to protect against UV radiation-induced elastin promoter activation. In addition, in vivo studies demonstrated that gluconolactone treatment does not result in a significant increase in sunburn cells. Further investigation of this and other PHAs is necessary to identify their potential role in preventing and repairing cutaneous photodamage. PMID- 14756650 TI - Watchmaker's pin-vise for manual tattooing of vitiligo. AB - BACKGROUND: Tattooing is a commonly used procedure for camouflage of depigmented lesions, especially in vitiligo. OBJECTIVE: To find a cheap and effective instrument for manual tattooing. METHODS: A watchmaker's pin-vise has been described for use as an instrument along with sewing needles for tattooing in vitiligo. RESULTS: A watchmaker's pin-vise is a cheap and effective for manual tattooing of vitiligo. CONCLUSION: A watchmaker's pin-vise loaded with sewing needles is an effective, cheap, and sterilizable instrument for tattooing. Sewing needles can be used as disposable needles. PMID- 14756651 TI - The use of the melolabial Burow's graft in the reconstruction of combination nasal sidewall-cheek defects. AB - BACKGROUND: Combination defects involving both the nasal sidewall or ala and cheek can be challenging to reconstruct. METHODS: Patients with skin cancers involving the nasal sidewall or ala and medial cheek treated with Mohs micrographic surgery were reconstructed using a cheek advancement flap in combination with a melolabial Burow's graft. RESULTS: This technique provides a simple, reproducible, and cosmetically excellent method of repairing these defects. CONCLUSION: The use of this repair provides superb tissue matches and maintains natural aesthetic subunits, yielding an excellent result with minimal morbidity. PMID- 14756652 TI - Modified surgical excision for the treatment of chondrodermatitis nodularis. AB - BACKGROUND: Chondrodermatitis nodularis (CN) is a common inflammatory condition of the ear that produces a painful papule or nodule on the helix or antihelix. Excision of the cartilage alone has been demonstrated to be therapeutically and cosmetically effective. OBJECTIVE: To simplify the surgical procedure. METHOD: We used two steps to refine the classic technique. We excised a narrow ellipse of skin over the nodule and replaced cold steel dissection by injecting normal saline into the plane of cleavage between the skin and cartilage. RESULTS: These two refinements resulted in maintaining the clinical and cosmetic efficacy and simplified the surgical technique. CONCLUSION: The addition of an elliptical excision over the nodule and the use of hydrodissection in the surgical treatment of chondrodermatitis nodularis were of benefit to both the patient and the surgeon. PMID- 14756653 TI - Lower lip hypertrophy secondary to port-wine stain: combined surgical and carbon dioxide laser treatment. AB - BACKGROUND: Port-wine stains (PWSs) are capillary malformations that usually show progressive stasis of the vascular channels and cause slow hyperplasia of the soft and hard tissues. When these lesions involve the lower lip, macrocheilia may be developed along the time. Vascular-specific lasers are not adequate to correct these three-dimensional tissue deformities, and surgical management becomes necessary, resulting in considerable morbidity and aesthetic disturbances. OBJECTIVE: To report a case of macrocheilia secondary to PWS treated by combination of surgery and carbon dioxide laser. METHODS: A 51-year-old man with macrocheilia of the lower lip and severe functional impairment, secondary to long evolution PWS, received treatment with carbon dioxide laser vaporization and minimal surgical correction, resulting in significant improvement of the lower lip hypertrophy, good aesthetic and functional status, and preservation of the muscular function. CONCLUSIONS: Combined carbon dioxide laser and surgery treatment may constitute a valuable alternative in treatment of macrocheilia secondary to PWS because bleeding risk is minimized and improves the preservation of muscular function and aesthetic results in relationship to conventional surgical approaches. PMID- 14756654 TI - Verrucous hyperplasia of the great toe: a case and a review of the literature. AB - BACKGROUND: Verrucous hyperplasia is a hyperplastic disease that manifests as coalescent warty papules at stump amputation sites. Often overlooked, it can be confused with verrucous carcinoma. As such, it is of concern for dermatologic surgeons. OBJECTIVE: To present a patient with verrucous hyperplasia and review its literature. METHODS: We report a 62-year-old diabetic patient with a verrucous nodule that arose at the amputation site of her first three right toes. Despite repeated surgical removals and skin graftings, the verrucous nodule redeveloped. A biopsy revealed digitate epidermal hyperplasia with dilated tortuous capillaries in thin dermal papilla without invasive features, mitotic figures, or acanthotic downgrowth. Polymerase chain reaction did not detect human papilloma virus. RESULTS: Verrucous hyperplasia secondary to amputation was diagnosed. CONCLUSION: Verrucous hyperplasia occurs should not be confused with verrucous carcinoma or warts. Moreover, because it recurs after removal, surgery is not indicated; rather, compression therapy is indicated. PMID- 14756655 TI - Squamous cell carcinoma of the nail unit with evidence of bony involvement: a multidisciplinary approach to resection and reconstruction. AB - BACKGROUND: Squamous cell carcinoma (SCC) of the nail bed is a rare disorder that is often misdiagnosed for years before definitive diagnosis with biopsy. Proper evaluation of this carcinoma includes radiographic evaluation for bony involvement of the phalanges of the affected digit. If bony involvement is evident by x-ray, amputation of the distal phalanx or the affected digit is warranted. Mohs micrographic surgery of the affected nail unit is advantageous in preserving vital tissue for reconstruction after phalangeal amputation by a hand surgeon, thus maximizing preservation of the densely innervated tissue from the volar finger pulp. This approach may maximize functional capacity of the reconstructed digit. OBJECTIVE: To describe a multidisciplinary approach to resection, amputation, and reconstruction of digits with SCC of the nail bed. METHODS: This is a description of three cases and a review of the pertinent medical literature. RESULTS: Three SCCs of the nail bed were excised with Mohs micrographic surgery, preserving the volar pulp and skin of the distal finger. The patients were then referred for distal phalanx amputation and reconstruction by a hand surgeon. All patients remained disease free with acceptable function of the reconstructed digits at 15, 17, and 38 months of follow-up. CONCLUSION: Although uncommon, SCC of the nail bed must be considered in all nails with chronic disease. Preoperative evaluation should include hand radiographs in search of bony involvement. Tissue-sparing excision combined with distal amputation of the affected phalanx and reconstruction of the digit using spared tissue may maximize hand and digit function. PMID- 14756656 TI - Eyelid sebaceous carcinoma masquerading as in situ squamous cell carcinoma. AB - BACKGROUND: Sebaceous carcinoma (SC) accounts for 1% to 5.5% of all eyelid malignancies. Diagnosis is often delayed because of its ability to masquerade as other periocular lesions both clinically and histologically. OBJECTIVE: To promote a high index of suspicion among Mohs surgeons for SC when managing biopsy proven in situ squamous cell carcinoma (SCC) of the eyelid, particularly the upper eyelid. METHODS: This is a retrospective case review. RESULTS: A 77-year old woman and a 71-year-old man were referred for Mohs micrographic surgery with a diagnosis of upper eyelid in situ SCC on biopsy. Intraoperatively, the presence of clear cell differentiation, in addition to extensive conjunctival involvement, leads to the correct diagnosis of SC. One case was subsequently found to have Muir Torre syndrome. CONCLUSION: Early diagnosis of eyelid SC leads to a better outcome and a higher survival rate. Hence, when managing in situ SCC of the eyelids, particularly the upper eyelid, Mohs surgeons should be aware of the possibility of SC and actively look for sebaceous cell differentiation or extensive conjunctival spread. PMID- 14756657 TI - Carcinoma en cuirasse presenting as keloids of the chest. AB - BACKGROUND: Carcinoma en cuirasse is a form of metastatic cutaneous breast malignancy occurring most commonly on the chest as a recurrence of breast cancer, but it can be the primary presentation. OBJECTIVE: To discuss the clinical features of carcinoma en cuirasse that distinguish it from hypertrophic scars and keloids of the chest. METHOD: We report a 63-year-old woman with primary cutaneous breast carcinoma presenting as keloid nodules on the chest that failed treatments for keloids. Biopsy revealed a pattern of breast carcinoma in the skin. RESULTS: After further workup, no tumor was found in the deep breast tissue, but metastases were found in her axillary lymph nodes. CONCLUSIONS: Unusual keloid-like nodules or scars on the chest that fail to respond to therapy may be primary or metastatic malignancies, and adequate histologic verification should be obtained to avoid delay in the proper treatment. PMID- 14756658 TI - Aggressive squamous cell carcinoma originating as a Marjolin's ulcer. AB - BACKGROUND: Marjolin's ulcer is an epidermoid carcinoma arising in a scar or chronic wound and can have an aggressive course. OBJECTIVE: To present a case of squamous cell carcinoma arising in a burn scar with resulting metastases and to discuss Marjolin's ulcer. RESULTS: The patient continued to have further metastatic disease despite aggressive surgical treatment. CONCLUSION: In following patients with chronic ulcers and wounds, it is important to evaluate any changes immediately with biopsies and further imaging studies if indicated in order to treat effectively. Even aggressive surgical intervention will sometimes be inadequate in treating these tumors. PMID- 14756659 TI - Mohs micrographic surgery for the eradication of phaeohyphomycosis of the hand. AB - BACKGROUND: Phaeohyphomycosis is a rare mycotic infection that is caused by dematiaceous fungi requiring surgical excision or long-term use of oral antifungal agents for treatment. OBJECTIVE: To report a case of phaeohyphomycosis of the dorsal hand successfully cleared with Mohs micrographic surgery. METHODS: We performed Mohs micrographic surgery on phaeohyphomycosis of the dorsal hand. The fungus was cleared in three stages of surgery. Permanent processing and special stains of the final stage confirmed eradication of the infection. RESULTS: The patient remained free of the phaeohyphomycosis, without complications, at the 6-month follow-up. CONCLUSION: Mohs micrographic surgery is an effective, tissue-sparing technique for the eradication of phaeohyphomycosis, potentially eliminating the need for costly long-term antifungal therapy. PMID- 14756660 TI - Secondary mucinous carcinoma of the skin: metastatic breast cancer. AB - BACKGROUND: Breast cancer is the most common cancer in women. Its involvement of skin is the most frequent of visceral cancers in women. In cutaneous metastatic disease, including breast cancer, the clinical and histologic pattern may be specific or nonspecific. Specific clinical patterns of cutaneous metastatic disease are linked with breast cancer but occur less often with other cancers metastatic to skin. Likewise, specific histologic patterns of cutaneous metastatic disease are linked with breast cancer but occur less often with other cancers metastatic to skin. OBJECTIVE: To present a case of a mucinous breast cancer metastatic to skin where the histologic pattern is similar to the primary tumor. METHODS: This is a case report and a literature review. RESULTS: Metastatic breast cancer may rarely resemble primary skin cancer, in this case primary mucinous carcinoma of the skin. We describe a 60-year-old woman with breast cancer with the incidental finding of a nonspecific, soft, solitary nodule on her back. It was found to contain mucinous material and on close examination was found to be a metastatic mucinous carcinoma of the skin from a primary adenocarcinoma of the breast. CONCLUSION: One usually considers that hard, firm nodules are more suggestive of cutaneous metastatic disease than soft, nondescript ones, but one should be careful to consider secondary mucinous carcinoma of the skin and a histologically similar solitary cutaneous metastasis. PMID- 14756661 TI - Re.: Cosmetic surgery is not plastic surgery. PMID- 14756662 TI - Modified purse string suture closure. PMID- 14756663 TI - Preauricular tubed pedicle flap repair of a superior antihelical defect. PMID- 14756664 TI - Taking it to the max: the genetic and developmental mechanisms coordinating midfacial morphogenesis and dysmorphology. AB - The rapid proliferative expansion and complex morphogenetic events that coordinate the development of the face underpin the sensitivity of this structure to genetic and environmental insult and provide an explanation for the high incidence of midfacial malformation. Most notable of these malformations is cleft lip with or without cleft palate (CLP) that, with an incidence of between one in 600 and one in 1000 live births, is the fourth most common congenital disorder in humans. Despite the obvious global impact of the disorder and some recent progress in identifying causative genes for some prominent syndromal forms, our knowledge of the key genetic factors contributing to the more common isolated cases of CLP is still remarkably patchy. The current understanding of the molecular and cellular processes that orchestrate morphogenesis of the midface, with emphasis on events leading to fusion of the lip and primary palate, is detailed in this review. The roles of crucial factors identified from relevant animal model systems, including BMP4 and SHH, and the likely events perturbed by key genes pinpointed in human studies [such as PVRL1, IRF6p63, MID1, MSX1, and PTCH1] are discussed in this light. New candidates for human CLP genes are also proposed. PMID- 14756668 TI - Concordance of phenotypic expression and gender identity in a large kindred with a mutation in the androgen receptor. AB - A 14-year-old female presented to the Pediatric Endocrine Clinic, Universidade Federal o Parana Curitiba, Brazil, for obesity. A few years later, despite normal breast development, the patient had failed to menstruate and lacked pubic and axillary hair. Laboratory analyses revealed high levels of testosterone. Karyotype analysis was XY. Direct sequencing of her genomic DNA showed a G to T transition at nucleotide 2089 at exon 2 in the androgen receptor gene, resulting in a substitution of Phe for Cys at position 576. This mutation disrupts the first Zn finger critical to DNA binding and transcriptional activity and results in complete androgen-insensitivity syndrome (CAIS). This individual was part of 700-member multigenerational kindred of German origin living in small villages in Southern Brazil. Family members who gave informed consent were screened using a polymerase chain reaction-based method. Nineteen CAIS-affected individuals and carriers were identified. All presented with infertility and lack of or sparse pubic hair. The prevalence of common AIS within the kindred greatly exceeds that of the general population and is due in part to their isolated familial and community structures. All individuals are genuinely feminine in their appearance, sex behavior, gender identity, and integration within their communities. We conclude that CAIS leads to complete feminization of XY individuals and results in individuals who are psychologically and socially established and integrated as women within the familial and cultural contexts of their communities. PMID- 14756669 TI - Polymorphisms in cytokines and growth factor genes and their association with acute rejection and recurrence of hepatitis C virus disease in liver transplantation. AB - Acute rejection (AR) and recurrence of hepatitis C virus (HCV) infection are complications after liver transplantation (LTx). Genetic factors play a role in cytokine production as a consequence of polymorphisms within cytokine genes. Our goal was to identify genetic factors that might be associated with AR and recurrence of HCV in liver transplant recipients (LTxRs). We studied 77 Caucasian LTxRs and 100 Caucasian healthy individuals. We studied single-nucleotide polymorphisms (SNPs) in tumor necrosis factor-alpha[TNF-alpha, interleukin-6 (IL 6), IL-10, transforming growth factor-beta1, and angiotensin-converting enzyme genes by SNaPSHOT trade mark Multiplex assay. SNPs were classified as high producers (HP), intermediate producers (IP), or low producers (LP), and their association with AR and recurrence of HCV were studied. The frequency of TNF alpha IP and HP genotypes was significantly higher in LTxRs with AR in comparison to patients without AR (TNF-alpha HP -238: 63 vs 20%, p < 0.001; TNF-alpha HP 308: 47.4 vs 20%, p = 0.02). The frequency of IL-6 IP and HP genotypes was higher in patients with AR episodes, but the difference was not statistically significant (p = 0.14). However, when we analyzed the simultaneous presence of pro-inflammatory genotypes in the same patient, we found a significant difference between patients with and without AR, respectively (42.1 vs 14.6%, p = 0.012). Moreover, the frequency of the IL-10 LP genotype was higher in LTx patients with AR (p = 0.001) compared to patients without AR. There was an association between pro-inflammatory genotypes and HCV recurrence. Our data suggest that cytokine gene polymorphisms might play a role in AR and HCV recurrence in LTxRs. PMID- 14756670 TI - The Delta>15 Kb deletion French Canadian founder mutation in familial hypercholesterolemia: rapid polymerase chain reaction-based diagnostic assay and prevalence in Quebec. AB - Approximately one in 500 individuals in Western population has autosomal dominant familial hypercholesterolemia due to mutations in the low-density lipoprotein receptor (LDLR) gene. Screening for these mutations is hampered by their large number, except in founder populations. We identified the breakpoint of the >15 kb deletion involving the LDLR gene promoter and exon 1, responsible for more than 60% of French Canadian hypercholesterolemia cases, as well as the breakpoint of the 5 kb deletion of exons 2 and 3 that accounts for an additional 5% of cases. Both deletions appear to be because of homologous recombination by unequal crossing-over between the left arms of Alu repeats. Using RepeatMasker, we determined that 55% of the LDLR gene is composed of Alu elements; thus, it is not surprising that most LDLR rearrangements involve at least one Alu. Furthermore, we developed a rapid polymerase chain reaction-based assay for the French Canadian-1 (>15 kb) and French Canadian-5 (5 kb) hypercholesterolemia alleles. Screening a representative population sample of 943 French Canadian youths whose LDL cholesterol levels were above the 50th percentile allowed us to estimate the prevalence of the >15 kb allele as 0.11% (95% confidence interval, 0.03-0.38). PMID- 14756671 TI - Genetic testing in spinocerebellar ataxia in Taiwan: expansions of trinucleotide repeats in SCA8 and SCA17 are associated with typical Parkinson's disease. AB - DNA tests in normal subjects and patients with ataxia and Parkinson's disease (PD) were carried out to assess the frequency of spinocerebellar ataxia (SCA) and to document the distribution of SCA mutations underlying ethnic Chinese in Taiwan. MJD/SCA3 (46%) was the most common autosomal dominant SCA in the Taiwanese cohort, followed by SCA6 (18%) and SCA1 (3%). No expansions of SCA types 2, 10, 12, or dentatorubropallidoluysian atrophy (DRPLA) were detected. The clinical phenotypes of these affected SCA patients were very heterogeneous. All of them showed clinical symptoms of cerebellar ataxia, with or without other associated features. The frequencies of large normal alleles are closely associated with the prevalence of SCA1, SCA2, MJD/SCA3, SCA6, and DRPLA among Taiwanese, Japanese, and Caucasians. Interestingly, abnormal expansions of SCA8 and SCA17 genes were detected in patients with PD. The clinical presentation for these patients is typical of idiopathic PD with the following characteristics: late onset of disease, resting tremor in the limbs, rigidity, bradykinesia, and a good response to levodopa. This study appears to be the first report describing the PD phenotype in association with an expanded allele in the TATA-binding protein gene and suggests that SCA8 may also be a cause of typical PD. PMID- 14756672 TI - Hereditary non-polyposis colorectal cancer and the role of hPMS2 and hEXO1 mutations. AB - Hereditary non-polyposis colorectal cancer (HNPCC) is an autosomal dominant, inherited condition that is characterized primarily by the development of early onset colorectal cancer and a number of other epithelial malignancies. The underlying genetic basis of the disease is associated with a breakdown of DNA mismatch repair. There are many genes involved in DNA-mismatch repair, and five of them have been implicated in HNPCC. Two of the genes (hMSH2 and hMLH1) account for the majority of HNPCC families (approximately 60%), and it is not known what the exact contributions of the remaining three genes (hPMS1, hPMS2, and hMSH6) are in relation to this condition. In addition, a sixth gene (hEXO1) has been associated with a disease phenotype that is consistent with HNPCC. Current estimates suggest that all four of these genes, combined, may account for up to 5% of families. In this report, we examine the contribution of hPMS2 and hEXO1 to a well-defined set of families that fulfill the diagnostic criteria for HNPCC. The genes, hPMS2 and hEXO1, were studied by denaturing high performance liquid chromatography (DHPLC) analysis in 21 families that have previously been determined not to have mutations in hMSH2 or hMLH1. hPMS2 accounts for a small proportion of HNPCC families, and none were deemed to be associated with hEXO1. Mutations in hPMS2 appear to account for a small proportion of families adhering to the Amsterdam II criteria, whereas hEXO1 does not appear to be associated with HNPCC. PMID- 14756673 TI - The clinical picture of the Borjeson-Forssman-Lehmann syndrome in males and heterozygous females with PHF6 mutations. AB - The usual description of the Borjeson-Forssman-Lehmann syndrome (BFLS) is that of a rare, X-linked, partially dominant condition with severe intellectual disability, epilepsy, microcephaly, coarse facial features, long ears, short stature, obesity, gynecomastia, tapering fingers, and shortened toes. Recently, mutations have been identified in the PHF6 gene in nine families with this syndrome. The clinical history and physical findings in the affected males reveal that the phenotype is milder and more variable than previously described and evolves with age. Generally, in the first year, the babies are floppy, with failure to thrive, big ears, and small external genitalia. As schoolboys, the picture is one of learning problems, moderate short stature, with emerging truncal obesity and gynecomastia. Head circumferences are usually normal, and macrocephaly may be seen. Big ears and small genitalia remain. The toes are short and fingers tapered and malleable. In late adolescence and adult life, the classically described heavy facial appearance emerges. Some heterozygous females show milder clinical features such as tapering fingers and shortened toes. Twenty percent have significant learning problems, and 95% have skewed X inactivation. We conclude that this syndrome may be underdiagnosed in males in their early years and missed altogether in isolated heterozygous females. PMID- 14756674 TI - Effective long-term control of cardiac events with beta-blockers in a family with a common LQT1 mutation. AB - The congenital long QT syndrome (LQTS) is characterized by a prolonged QT interval on the surface electrocardiogram and an increased risk of recurrent syncope and sudden cardiac death. Mutations in seven genes have been identified as the molecular basis of LQTS. beta-blockers are the treatment of choice to reduce cardiac symptoms. However, long-term follow-up of genotyped families with LQTS has been rarely reported. We have clinically followed a four-generation family with LQTS being treated with beta-blocker therapy over a period of 23 years. Seven family members were carriers of two amino acid alterations in cis (V254M-V417M) in the cardiac potassium channel gene KCNQ1. Voltage-clamp recordings of mutant KCNQ1 protein in Xenopus oocytes showed that only the V254M mutation reduced the IKs current and that the effect of the V417M variant was negligible. The family exhibited the complete clinical spectrum of the disease, from asymptomatic patients to victims of sudden death before beta-blocker therapy. There was no significant reduction in QTc (556 +/- 40 ms(1/2) before therapy, 494 +/- 20 ms(1/2) during 17 years of treatment; n = 5 individuals). Of nine family members, one female died suddenly before treatment, three females of the second generation were asymptomatic, and four individuals of the third and fourth generation were symptomatic. All mutation carriers were treated with beta blockers and remained asymptomatic for a follow-up up to 23 years. Long-term follow-up of a LQT1 family with a common mutation (V254M) being on beta-blocker therapy was effective and safe. This study underscores the importance of long term follow-up in families with specific LQT mutations to provide valuable information for clinicians for an appropriate antiarrhythmic treatment. PMID- 14756675 TI - Supernumerary marker 15 chromosome in a patient with Prader-Willi syndrome. PMID- 14756676 TI - Heterozygous manifestations in female carriers of Mal de Meleda. PMID- 14756677 TI - Importance of calcitonin gene-related peptide, adenosine and reactive oxygen species in cerebral autoregulation under normal and diseased conditions. AB - 1. Mechanisms regulating cerebral circulation, including autoregulation of cerebral blood flow (CBF), have been widely investigated. Vasodilators such as nitric oxide, prostacyclin, calcitonin gene-related peptide (CGRP) and K+ channel openers are well known to have important roles in the physiological and pathophysiological control of CBF autoregulation. In the present review, the focus is on the mechanism(s) of altered CBF autoregulation after traumatic brain injury and subarachnoid haemorrhage (SAH) and on the effect of adenovirus mediated transfer of Cu/Zn superoxide dismutase (SOD)-1 in amelioration of impaired CBF autoregulation. 2. The roles of CGRP and adenosine are particularly emphasized, both being implicated in the autoregulatory vasodilation of the pial artery in response to hypotension. 3. After fluid percussion injury, production of NADPH oxidase-derived superoxide anion and activation of tyrosine kinase links the inhibition of K+ channels to impaired autoregulatory vasodilation in response to acute hypotension and alterations in CBF autoregulation in rat pial artery. 4. Subarachnoid haemorrhage during the acute stage causes an increase in NADPH oxidase-dependent superoxide formation in cerebral vessels in association with activated tyrosine phosphorylation-coupled increased expression of gp91phox mRNA and membrane translocation of Rac protein, thereby resulting in a significant reduction of autoregulatory vasodilation. 5. Fluid percussion injury and SAH induced overproduction of superoxide anion in cerebral vessels contributes to the impairment of CBF autoregulation and administration of recombinant adenovirus mediated transfer of the Cu/Zn SOD-1 gene effectively ameliorates the impairment of CBF autoregulation of the pial artery. PMID- 14756679 TI - Respiratory and behavioural compensation during chronic severe loading in a hypoxic rat model. AB - 1. Respiratory load compensation plays an important role in the maintenance of an appropriate level of ventilation. We investigated the ventilatory and behavioural responses to a severe load causing both hypercapnia and hypoxaemia in a rat model. 2. A thin silicone tube (15 mm in length, 1 mm ID) was implanted surgically in the trachea. Arterial blood gases and ventilatory and motor activities were assessed serially and non-invasively over a period of 8 weeks. 3. Ventilatory loading produced severe respiratory acidosis with concomitant hypoxaemia immediately after imposition of the load, but there was a considerable improvement of arterial blood gases at 1 day after the start of respiratory loading. Ventilatory loading also caused a gradual increase in ventilatory activity, requiring 7 days to reach the maximum level. There was a reduction in daily motor activity immediately after the ventilatory loading, but this reduction recovered gradually, together with recovery of reduced food and water intake. Administration of a hyperoxic gas mixture during the ventilatory loading decreased ventilatory activity while improving reduced motor activity. 4. These observations suggest that severe chronic ventilatory loading may induce adaptive responses that compensate rapidly for the disturbed acid-base balance with slow and gradual increases in ventilatory activity while matching the increase in motor activity/metabolic rate. PMID- 14756678 TI - An antidiabetic thiazolidinedione induces eccentric cardiac hypertrophy by cardiac volume overload in rats. AB - 1. To assess the involvement of volume overload in the development of cardiac hypertrophy during treatment with an antidiabetic thiazolidinedione, changes in cardiac anatomy and parameters of cardiac volume overload were evaluated in female Sprague-Dawley rats treated with the thiazolidinedione derivative T-174. 2. Two week administration of T-174 (13 and 114 mg/kg per day) increased absolute and relative heart weights by 11-24%, demonstrating the development of cardiac hypertrophy. There was no evidence of oedema in hearts from treated rats. 3. Both plasma and blood volumes were increased in T-174-treated rats without any changes in systolic blood pressure and heart rate, whereas haematocrit was decreased. In accordance with the existence of volume overload, both left ventricular end diastolic pressure and right atrial pressure were increased. Morphometric analysis of hearts revealed that T-174 induced eccentric heart hypertrophy, as characterized by a small increase in wall thickness and a large increase in the chamber volume, which is characteristic of volume overload. Volume overload is suggested as the possible trigger mechanism because blood volume expansion preceded cardiac hypertrophy and there was a high correlation between heart weight and blood volume. 4. T-174-treated streptozotocin-induced diabetic rats also exhibited blood volume expansion and cardiac hypertrophy. 5. These findings suggest that cardiac volume overload is induced by plasma volume expansion and contributes to the development of eccentric cardiac hypertrophy during treatment with antidiabetic thiazolidinediones. Although thiazolidinediones are insulin sensitizing agents, these cardiac effects are likely to be mediated independently of insulin. PMID- 14756680 TI - Dose-dependent effect of alcohol on insulin-like growth factor systems in male rats. AB - 1. Chronic alcohol treatment has been reported to be associated with liver and kidney damage. The insulin-like growth factor (IGF) is the major growth factor related to alcohol consumption. However, the effect of alcohol on the IGF system in the liver and kidney has not been fully elucidated. Thus, the present study was conducted to investigate this issue. 2. Alcohol reduced the level of IGF-I in a dose-dependent manner in the serum, liver and kidney. Alcohol also decreased the level of IGF-II in the liver. In contrast, alcohol increased the level of IGF II in the serum and kidney. These observations were correlated with IGF-I and IGF II mRNA expression in the liver and kidney. 3. To examine the effect of alcohol on IGF receptors in the liver and kidney, IGF-I receptor mRNA was measured. Alcohol decreased IGF-I receptor mRNA in the liver and kidney. 4. In experiments performed to examine the regulation of IGF-binding proteins (IGFBP), alcohol increased serum levels of IGFBP-1. However, alcohol had no effect on serum levels of IGFBP-2, -3 and -4. These effects were also observed in the liver and kidney. 5. In conclusion, alcohol alters the IGF system in rat liver and kidney in a tissue-specific manner, which may contribute to the metabolic dysfunction following chronic alcohol consumption. PMID- 14756681 TI - Renal excretion mechanisms of the antiviral nucleoside analogue AM 188 in the rat isolated perfused kidney. AB - 1. AM 188 is an antiviral guanosine analogue that undergoes extensive renal excretion in humans. The present study was designed to investigate the disposition of AM 188 over a range of concentrations in the rat isolated perfused kidney (IPK) to explore the mechanisms involved in its renal handling. 2. Right kidneys of male Sprague-Dawley rats (n = 23) were isolated and perfused in recirculating mode with Krebs'-Henseleit (pH 7.4) buffer containing 0.65% bovine serum albumin, 3.6% dextran, amino acids and glucose. [14C]-Inulin was added to the perfusate reservoir to permit estimation of glomerular filtration rate (GFR). [3H]-AM 188 and unlabelled AM 188 were added to the perfusate as a bolus initially, followed by a constant rate of infusion at 5, 25, 125, 500 or 1000 microg/min to achieve initial target perfusate concentrations of 1, 5, 25, 100 or 200 microg/mL, respectively. During the 130 min over which AM 188 was infused, urine was collected in 10 min intervals (commencing 10 min after the bolus dose) and perfusate was collected at the mid-point of these intervals to permit calculation of the renal clearance (CLR) of AM 188. Binding of AM 188 in perfusate, measured using ultrafiltration, was negligible. 3. The bolus dose and infusion regimen produced relatively stable AM 188 concentrations in perfusate in the 5, 25 and 125 micro g/min groups and progressively increasing concentrations in the 500 and 1000 microg/min groups. High-pressure liquid chromatography analysis of IPK perfusate and urine suggested that there was no or negligible metabolism of AM 188 in the kidney. The CLR/GFR ratio for AM 188 (mean+/-SD) was 5.76 +/- 1.57, 5.99 +/- 0.52, 6.02 +/- 1.47, 3.38 +/- 0.26 and 1.08 +/- 0.42 in the 5, 25, 125, 500 and 1000 microg/min groups, respectively, showing significant reductions at the two highest infusion rates (P < 0.05). Although there was no difference between the five groups in the distribution of AM 188 between kidney tissue and perfusate (KT/P), at the end of perfusion the corresponding urine-to tissue concentration ratio declined significantly in the 1000 microg/min group. 4. AM 188 undergoes substantial net renal secretion over a wide range of perfusate concentrations. A reduction in renal clearance at perfusate concentrations above 25 microg/mL could be due to saturation of carrier-mediated transport at the brush border membrane and/or a solubility limitation leading to precipitation of AM 188 in tubular cells and/or tubular urine. PMID- 14756682 TI - Effect of renal perfusion pressure on responses of intrarenal blood flow to renal nerve stimulation in rabbits. AB - 1. We investigated how sympathetic nerve activity and renal perfusion pressure (RPP) interact in controlling renal haemodynamics in pentobarbitone-anaesthetized rabbits. 2. Renal blood flow (RBF) was reduced by electrical renal nerve stimulation (0.5-8 Hz), with RPP set using an extracorporeal circuit to 65, 100 and 135 mmHg. 3. Responses of RBF and cortical laser Doppler flux to renal nerve stimulation were blunted by increased RPP. For example, 4 Hz stimulation reduced RBF by 68 +/- 7% with baseline perfusion pressure approximately 65 mmHg, but only by 22 +/- 3% at approximately 135 mmHg. Medullary laser Doppler flux was less responsive than cortical laser Doppler flux to renal nerve stimulation and its response was not dependent on perfusion pressure. 4. When perfusion pressure was clamped at its baseline level during renal nerve stimulation, responses of RBF and cortical laser Doppler flux, but not medullary laser Doppler flux, were still blunted with increased baseline perfusion pressure. 5. A frequency rich stimulus was applied to assess the effects of perfusion pressure on dynamic neural control of RBF. Renal blood flow responded similarly at each level of perfusion pressure, as a low-pass filter with a pure time delay. 6. Our results suggest that, in the rabbit extracorporeal circuit model, increased RPP blunts the ability of steady state renal nerve stimulation to reduce cortical, but not medullary perfusion. However, in this model the level of RPP appears to have little impact on dynamic neural control of RBF. PMID- 14756683 TI - Nitric oxide promotes mitogen-induced dna synthesis in human dermal fibroblasts through cGMP. AB - 1. Nitric oxide (NO) is a free radical with multiple functions in cellular pathophysiology. Nitric oxide has been proven to play an important role in wound healing; however, the mechanisms by which NO may promote wound healing are not clearly understood. We have investigated the effect of NO on growth factor induced DNA synthesis in human dermal fibroblasts to suggest interactions between growth factors and NO as a possible mechanism for the role of NO in wound healing. 2. The NO donor sodium nitroprusside (SNP) significantly (P < 0.001) increased fetal bovine serum-induced thymidine incorporation into the DNA of human dermal fibroblasts. The maximal comitogenic concentration of SNP (100 micro mol/L) was also found to significantly (twofold; P < 0.01) enhance fibroblast growth factor- or platelet-derived growth factor-induced DNA synthesis. 3. Nitric oxide treatment significantly increased the production of cGMP. 8-Bromo-cGMP, a stable structural analogue of cGMP, was found to markedly potentiate (P < 0.001) the growth factor-induced DNA synthesis. 4. This study concludes that NO and cGMP promote growth factor-induced DNA synthesis in dermal fibroblasts, suggesting another possible mechanism by which NO may promote skin wound healing. PMID- 14756684 TI - Cardiovascular responses to orthostatic and other stressors in men and women are independent of sex. AB - 1. Cardiovascular responses to the stress of orthostasis, forearm (FA) ischaemia (reactive hyperaemia) and FA exercise (postexercise hyperaemia) are well described. Although sex differences in responses to orthostatic stress have been reported, few studies have examined the impact of sex on reactive hyperaemia and none has commented with regard to postexercise hyperaemia. 2. We investigated 11 men (mean (+/-SEM) age 18.5 +/- 0.3 years) and 10 women (18.8 +/- 0.8 years), all of whom were sedentary, with women being studied in the mid-follicular phase of their menstrual cycle. We measured blood pressure (BP), heart rate (HR) and forearm blood flow (FBF) in response to a fixed sequence of orthostatic, ischaemic and exercise stressors. 3. Orthostatic stress (10 min at -50 mmHg lower body negative pressure; LBNP) induced presyncopal signs in one man and three women. In all other subjects, BP was well maintained, with FBF decreasing and HR increasing similarly in both sexes. The tachycardia was earlier in onset in men and reached significantly higher absolute levels in women during the final 5 min of LBNP, but the percentage changes and integrated responses of both HR and FBF were not different between sexes. 4. The increases in FBF following either 10 min FA ischaemia or 10 min FA exercise were similar in men and women in terms of peak blood flow, percentage change, rate of recovery and total blood flow response. 5. In conclusion, although women were less tolerant of orthostatic stress than men, the cardiovascular responses to this and the other stressors appeared essentially independent of sex. PMID- 14756685 TI - Oxcarbazepine antinociception in animals with inflammatory pain or painful diabetic neuropathy. AB - 1. Diabetic neuropathy is one of the most frequent complications of diabetes mellitus. However, the mechanisms underlying these disorders are not yet well defined and it has been reported that currently available analgesics have hardly any ameliorating effect on painful diabetic neuropathy. 2. The purpose of the present study was to evaluate the antinociceptive effect of oxcarbazepine (OCBZ), a keto derivative of carbamazepine (CBZ), in animal models generally used in pain research and in rats and mice with streptozotocin (STZ)-induced diabetes. In addition, we compared the effect of OCBZ with those of CBZ, mexiletine and morphine. 3. Diabetes was induced by injection of STZ at a dose of 300 mg/kg (i.p.) in mice and 50 mg/kg (i.v.) in rats. Experiments were conducted 2 weeks after STZ injection and those animals with a serum glucose level above 400 mg/dL were used for data analysis. Antinociceptive effects of the drugs were evaluated by the paw withdrawal test (normal, STZ-induced diabetic and carrageenin-injected rats), tail-flick test (normal and STZ-induced diabetic mice) and nociceptive behaviour (formalin-injected mice). 4. In the present study, diabetic mice showed thermal hyperalgesia and diabetic rats exhibited mechanical hyperalgesia. From these results, the STZ-induced diabetic animals used in the present study were found to be suitable for research on painful diabetic neuropathy. In STZ-induced diabetic animals, the antinociceptive effects of OCBZ, CBZ and mexiletine were facilitated, whereas the effect of morphine was attenuated, compared with effects in normal animals. 5. Oxcarbazepine inhibited the formalin-induced biphasic pain responses and increased the nociceptive threshold in the case of carrageenin induced hyperalgesia. In view of these results, inhibition of substance P mediated pain transmission may be involved in the antinociceptive action of OCBZ. 6. These results indicate that OCBZ has an analgesic action and is a possible therapeutic agent for the treatment of neuropathic pain, such as occurs in painful diabetic neuropathy. PMID- 14756686 TI - Cytotoxic effects of Coptis chinensis and Epimedium sagittatum extracts and their major constituents (berberine, coptisine and icariin) on hepatoma and leukaemia cell growth. AB - 1. The present study was conducted to evaluate the cytotoxic effects of Coptis chinensis and Epimedium sagittatum extracts and their major constituents on hepatoma and leukaemia cells in vitro. 2. Four human liver cancer cell lines, namely HepG2, Hep3B, SK-Hep1 and PLC/PRF/5, and four leukaemia cell lines, namely K562, U937, P3H1 and Raji, were used in the present study. 3. Of the two crude drugs, C. chinensis exhibited the strongest activity against SK-Hep1 (IC50 = 7 microg/mL) and Raji (IC50 = 4 microg/mL) cell lines. The IC50 values for C. chinensis on HepG2, Hep3B and PLC/PRF/5 cell lines were 20, 55 and 35 microg/mL, respectively. The IC50 values for C. chinensis on K562, U937 and P3H1 cell lines were 29, 29 and 31 microg/mL, respectively. 4. With the exception of HepG2 and Hep3B, the E. sagittatum extract inhibited the proliferation of all cell lines (SK-Hep1, PLC/PRF/5, K562, U937, P3H1 and Raji), with IC50 values of 15, 57, 74, 221, 40 and 80 microg/mL, respectively. 5. Interestingly, the two major compounds of C. chinensis, berberine and coptisine, showed a strong inhibition on the proliferation of both hepatoma and leukaemia cell lines, with IC50 values varying from 1.4 to 15.2 microg/mL and from 0.6 to 14.1 microg/mL, respectively. However, icariin (the major compound of E. sagittatum) showed no inhibition of either the hepatoma or leukaemia cell lines. 6. The results of the present study suggest that the C. chinensis extract and its major constituents berberine and coptisine possess active antihepatoma and antileukaemia activities. PMID- 14756687 TI - Age-dependent cardiovascular, renal and endocrine responses to furosemide in conscious lambs. AB - 1. The present study was performed to investigate some of the physiological responses to furosemide during postnatal maturation. 2. In 1- (n = 8) and 6-week old (n = 10) conscious, chronically instrumented lambs at least 3 days after surgery, three experiments were performed at intervals of 24-48 h and in random order. Various parameters of cardiovascular and endocrine function, as well as cumulative urinary flow rates, were measured before and after intravenous injection of 0 mg/kg (experiment one), 0.25 mg/kg (experiment two; low dose) and 5 mg/kg (experiment three; high dose) furosemide. 3. After high-dose furosemide, mean venous pressure decreased and there was a transient increase in mean arterial pressure in lambs aged 6 weeks. At 1 week of age, heart rate increased after high-dose furosemide and renal blood flow decreased. After high-dose furosemide, plasma renin activity increased in both groups of lambs, although the effects were greater in 1-week-old lambs. Plasma levels of arginine vasopressin increased after high-dose furosemide in lambs aged 1 but not 6 weeks. Cumulative urinary flow rate responses to furosemide were similar in 1- and 6-week-old lambs. 4. These data provide new information that cardiovascular and endocrine responses to furosemide are developmentally regulated. PMID- 14756688 TI - Effects of Hilsa ilisa fish oil on the atherogenic lipid profile and glycaemic status of streptozotocin-treated type 1 diabetic rats. AB - 1. The effects of oral administration of Hilsa (Hilsa ilisa) fish oil (1 g oil/kg bodyweight per day) on the lipid profile, platelet aggregation, anti-oxidative status and glycaemic control of streptozotocin (STZ; 90 mg/kg bodyweight)-treated type 1 diabetic rats were compared with those in fish oil-treated or untreated non-diabetic rats. 2. After 3 weeks of fish oil feeding, plasma total cholesterol decreased in both the non-diabetic and diabetic rats by 35 and approximately 10%, respectively, and triglyceride fell by 69 and 20%, respectively, compared with control rats. 3. Fish oil feeding decreased non-esterified fatty acids (NEFA) by 29% in diabetic rats but the NEFA level in non-diabetic rats was unaffected. 4. In non-diabetic and diabetic rats, platelet aggregation decreased by 49 and 37%, respectively, and total anti-oxidant status increased by 18 and 17%, respectively, after fish oil feeding. 5. Insulin levels increased by 27% in the fish oil-fed non-diabetic rats, whereas insulin levels were markedly decreased in diabetic rats. Glucose levels were not altered at all and fructosamine levels decreased by 29% only in fish oil-fed diabetic rats. 6. The results of the present study suggest that Hilsa ilisa fish oil may ameliorate the atherogenic lipid profile, platelet hyperaggregation and the anti-oxidative defence of STZ diabetic rats and the amelioration is thought to be independent of the effects of Hilsa on glycaemic control. PMID- 14756689 TI - Effect of caffeine on response of rabbit isolated corpus cavernosum to high K+ solution, noradrenaline and transmural electrical stimulation. AB - 1. Caffeine has wide-ranging activities on smooth muscles, including contractile and relaxant effects. The aim of the present study was to examine the activity of caffeine on rabbit corpus cavernosum (RCC). 2. The effects of caffeine (0.5-4.0 mmol/L) on the response of RCC to high K+ solution, noradrenaline (NA) and transmural electrical stimulation (EFS) were studied in a tissue bath system. 3. Caffeine did not contract the RCC. However, 0.5-4.0 mmol/L caffeine caused concentration-dependent relaxation of tension development in high-K+ (120 mmol/L) solution in contrast with the solvent control. At 4.0 mmol/L caffeine, high-K+ solution-induced tone of the RCC was reduced by 73.4 +/- 7.3%. Caffeine (0.5-4.0 mmol/L) also concentration-dependently relaxed NA (12.5 micro mol/L)-induced tonic contraction of the RCC. At 4.0 mmol/L caffeine, NA-induced tone of the RCC was reduced by 41.1 +/- 7.0%. Incubation of RCC in 2.0 mmol/L caffeine for 30 min prior to EFS (1-40 Hz) caused a marked rightward shift in the frequency-response curve. 4. The results of the present study suggest that caffeine exhibits relaxant activity on rabbit cavernosal smooth muscle and the mechanism of this activity possibly involves inhibition of Ca2+ signalling. PMID- 14756690 TI - Actions of the anti-oestrogen agent clomiphene on outward K+ currents in rat ventricular myocytes. AB - 1. The effects of clomiphene (CLM) on cardiac outward K+ current components from rat isolated ventricular myocytes were investigated using the whole-cell patch clamp technique. Clomiphene (10 micromol/L) significantly inhibited both peak (Ipeak) and end-pulse (Ilate) outward currents (elicited by a 500 msec voltage step from -40 to +50 mV in the presence of K+-containing intracellular and extracellular solutions) by approximately 37% (n = 6; P < 0.01) and 49% (n = 6; P < 0.01), respectively. In contrast, CLM had no effect on outward currents when K+ free solutions were used. 2. A double-pulse protocol and Boltzmann fitting were used to separate individual K+ current components on the basis of their voltage dependent inactivation properties. At potentials positive to -80 mV, two inactivating transient outward components (Ito) and (IKx) and a non-inactivating steady state component (Iss) could be distinguished. 3. Clomiphene inhibited both Ito and Iss. The maximal block of Ito and Iss induced by CLM (100 micromol/L) was approximately 61% (n = 5) and 43% (n = 5) with IC50 values of 1.54 +/- 0.39 and 2.2 +/- 0.4 micromol/L, respectively. In contrast, the peak magnitude of IKx was unaltered by CLM, although its time-course of inactivation was accelerated. 4. Further experiments whereby myocytes were superfused with the vasoactive peptide endothelin (ET)-1 (20 nmol/L) revealed that CLM (10 micro mol/L) completely abolished the ET-1-sensitive component of Iss. 5. Our findings demonstrate, for the first time, the effects of CLM on distinct cardiac K+ current components and show that CLM modulates the voltage-gated K+ current components Ito and IKx and inhibits the steady state outward current Iss in rat ventricular myocytes. PMID- 14756691 TI - Basal transcriptional regulation of rat AT1 angiotensin II receptor gene expression. AB - 1. Angiotensin II AT1A receptors are thought to play an important role in the development of hypertension. The transcriptional factor Sp1 is a ubiquitous transcriptional factor associated with GC-rich promoters and involved in basal promoter activity. 2. To examine basal transcriptional levels regulation of the rat AT1A receptor gene, we determined whether two GC-box-related regions within 100 bp of the rat AT1A receptor gene promoter are involved in the basal expression of the gene in A10 cells, a vascular smooth muscle cell line. 3. The electrophoretic mobility shift assay demonstrated that incubation of the -98/-79 region and -58/-34 region sequence oligonucleotides with nuclear extracts of rat hypothalamus, liver and adrenal formed DNA-protein complexes and that the addition of unlabelled oligonucleotides containing the Sp1 consensus sequence blocked the formation of the DNA-protein complex. The addition of antibody against Sp1 also blocked the formation of the DNA-protein complex. 4. The promoter/luciferase reporter assay demonstrated that the reporter activity of AT1A receptor promoters mutated either within the -98/-79 or the -58/-34 region was lower than that of intact AT1A receptor promoters. 5. The promoter activity of AT1A receptor promoters mutated within those two regions was lower than that of promoters mutated within either the -98/-79 or the -58/-34 region. 6. These findings suggest that GC-box-regulated sequences within the -98/-79 region and the -58/-34 region are additively involved in basal expression level of the AT1A receptor gene in A10 cells. PMID- 14756692 TI - Cardiorespiratory effects of intravenous N-methyl-D-aspartate challenge in anaesthetized rats. AB - 1. Experiments were performed to determine the effects of systemic application of N-methyl-d-aspartate (NMDA) on respiratory variables and blood pressure in 22 urethane/chloralose-anaesthetized, spontaneously breathing rats. 2. Bolus injection of NMDA at a dose of 27 micro mol/kg, i.v., in neurally intact rats evoked a depression of breathing, most apparent at 30 s, comprising a decrease in tidal volume (P < 0.001) and respiratory rate (P < 0.001). The expiratory apnoea appeared in three intact rats only. 3. The respiratory effects of NMDA were independent of the vagal integrity between lungs and the nodose ganglia. Elimination of supranodose connection to the medulla reduced the prolongation of the expiratory time (P < 0.01). 4. N-Methyl-d-aspartate induced an initial rise in blood pressure followed by hypotension in rats treated by infra- and supranodose vagotomy. 5. It is concluded that the respiratory response to systemic NMDA challenge occurs beyond lung vagi and indicates that neurons of the nodose ganglia contribute to NMDA inhibition of the expiratory time. PMID- 14756693 TI - Establishment of a new animal model of metabolic syndrome: SHRSP fatty (fa/fa) rats. AB - 1. We established a new animal model of metabolic syndrome, SHRSP fatty (fa/fa) rats, by crossing stroke-prone spontaneously hypertensive rats of the Izumo strain (SHRSP/Izm) to Zucker fatty (ZF) (fa/fa) rats. 2. The SHRSP fatty (fa/fa) rats have a missense mutation of the leptin receptor gene and plasma leptin concentrations are augmented. The SHRSP fatty (fa/fa) rats develop obesity and hypertension simultaneously. 3. Plasma metabolic parameters, including glucose, insulin and total cholesterol and triglyceride levels, were markedly elevated in SHRSP fatty (fa/fa) rats compared with SHRSP/Izm rats. Plasma triglyceride concentrations in SHRSP fatty (fa/fa) rats were significantly elevated compared with those in ZF (fa/fa) rats. The weight of adipose tissues in SHRSP fatty (fa/fa) rats was greater than that of SHRSP/Izm rats. The phenotype of SHRSP fatty (fa/fa) rats is similar to that of human metabolic syndrome. PMID- 14756694 TI - A < 1.7 cM interval is responsible for Dmo1 obesity phenotypes in OLETF rats. AB - 1. Dmo1 (Diabetes Mellitus OLETF type I) is a major quantitative trait locus for dyslipidaemia, obesity and diabetes phenotypes of male Otsuka Long Evans Tokushima Fatty (OLETF) rats. 2. Our congenic lines, produced by transferring Dmo1 chromosomal segments from the non-diabetic Brown Norway (BN) rat into the OLETF strain, have confirmed the strong, wide-range therapeutic effects of Dmo1 on dyslipidaemia, obesity and diabetes in the fourth (BC4) and fifth (BC5) generations of congenic animals. Analysis of a relatively small number of BC5 rats (n = 71) suggested that the critical Dmo1 interval lies within a < 4.9 cM region between D1Rat461 and D1Rat459. 3. To confirm the assignment of the Dmo1 critical interval, we intercrossed BC5 animals to produce a larger study population (BC5:F1 males; n = 406). For the present study, we used bodyweight at 18 weeks of age as an index of obesity; this phenotype is representative of the closely associated dyslipidaemia and hyperglycaemia phenotypes. 4. Interval mapping assigned logarithm of odds (LOD) peaks at the D1Rat90 marker (LOD = 9.11). One LOD support interval lies within the < 1.7 cM region between D1Rat461 and D1Rat459. 5. This large intercross study confirms that Dmo1 is likely localized within the interval. PMID- 14756695 TI - Detecting systematic bias between two raters. AB - 1. I recently reviewed, inter alia, methods for comparing two raters who make judgements on an ordered categorical scale, directed principally at the kappa statistic and its weaknesses. 2. The main weakness of the kappa statistic is that it fails to detect the all-important feature of systematic bias between raters. 3. I described various methods for detecting bias between two raters. These included a modified McNemar test and the single binomial test. Others that have been suggested are the symmetry of disagreement index (SD) and the marginal homogeneity test. 4. I now realize that none of the above four tests for bias is satisfactory, because all ignore the extent of agreement. 5. The bias index (BI) does take into account the extent of agreement, but its inventors did not propose how BI could be evaluated. I now describe a method for doing this. PMID- 14756697 TI - Methodological articles in addiction journals: do we need more, less, or about the same amount? PMID- 14756696 TI - Lipophilicity affects the pharmacokinetics and toxicity of local anaesthetic agents administered by caudal block. AB - 1. Drugs administered into the epidural space by caudal block are cleared by means of a process potentially affected by the lipophilic character of the compounds. 2. In the present study, we examined the relationship between the octanol-water partition coefficient (log Poct) and the time to reach the maximum plasma drug concentration (tmax) of lignocaine, bupivacaine and ropivacaine administered by caudal block in paediatric patients. We also examined the relationship between log Poct and the toxicity of these local anaesthetic agents in experimental models. The tmax and toxicity data were obtained from the literature. 3. Ropivacaine, with a log Poct of 2.9, exhibited a tmax of 61.6 min. The tmax of lignocaine, with a log Poct of 2.4, and bupivacaine, with a log Poct of with 3.4, were approximately 50% shorter than ropivacaine. At log Poct of approximately 3.0, the toxicity of these local anaesthetic agents was substantially increased. The relationship between log Poct and the convulsive effect in dogs was similar to the relationship between log Poct and the lethal dose in sheep. 4. With local anaesthetic agents, it appears that the relationship between log Poct and drug transfer from the epidural space to the blood stream is parabolic, being the slowest rate of transference at log Poct 3.0. Toxicity, due to plasma availability of these local anaesthetic agents, seems to be increased at log Poct equal or higher than 3.0 secondary to the highest transfer from plasma into the central nervous system. PMID- 14756699 TI - Can you have a long but rewarding addictions research career? PMID- 14756700 TI - Alcohol research as a career. PMID- 14756701 TI - Reflections on a career as an addictions researcher. PMID- 14756702 TI - Retrospections of a recovering drug researcher. PMID- 14756703 TI - Help the helper-addiction research and the helper syndrome. PMID- 14756704 TI - A few apologies, but no regrets. PMID- 14756705 TI - Big issues in unexpected places. PMID- 14756706 TI - The value of independence? PMID- 14756708 TI - Kettil Bruun Society for Social and Epidemiological Research on Alcohol. AB - The Kettil Bruun Society for Social and Epidemiological Research on Alcohol (KBS) was established in 1987 and is an independent organization open to all scientists working on problems related to social and epidemiological research on alcohol. The aim of the Society is to promote social and epidemiological research which fosters a comparative understanding of the social aspects of alcohol use and alcohol problems. In line with this the Society also aims at promoting a spirit of international collaboration. The Kettil Bruun Society is based on individual membership and, by 2003, has 197 fully paid-up members, representing 34 different countries over five continents. The main activities include an annual meeting as well as thematic meetings. In these meetings, discussions are emphasized by having precirculated papers and assigned discussants. The KBS also serves as a basis for organizing international collaborative projects. Project meetings or work-shops are often organized around the annual meetings, and the projects tend to run over several years. The Society's primary influence is through the mutual influence of its members on each others' thinking, the work of the projects that KBS sponsors and the influence its members have collectively on the development of the field. PMID- 14756709 TI - A postmarketing study of relative abuse liability of hypnotic sedative drugs. AB - AIMS: To demonstrate the utility of postmarketing studies using in-treatment drug and alcohol abusers as informants for assessing the relative abuse liability of sedative-hypnotic drugs. DESIGN: A survey was conducted that ascertained exposure to a variety of drugs with hypnotic/sedative properties and elicited subjective evaluations indicative of abuse liability. METHODS: Complete data were obtained from 297 admissions (78% male) to three addiction treatment sites in the United Kingdom. Subjects were asked 15 questions about 12 different drugs, including five benzodiazepines, three antidepressants, two non-benzodiazepine hypnotics and two antihistamines (plus one fictitious drug). Three of the benzodiazepines (diazepam, nitrazipam, temazepam) emerged as having substantially more abuse liability than any of the other drugs tested, as revealed by higher scores on abuse liability items (purchased on street, taken to get high, like drug, potential for addiction to drug). The antihistamines (chlorpheniramine, diphenhydramine) had lowest abuse liability profiles, while the antidepressants (amitriptyline, fluoxetine, trazadone) and non-benzodiazepine hypnotics (zolpidem, zopiclone) had similar profiles. CONCLUSION: This pilot study suggests that postmarketing information on hypnotic drug use obtained from drug addicts entering treatment produces data consistent with other measures of abuse liability. The data suggest that the risk of misuse of newer non-benzodiazepine hypnotics may be less than that of benzodiazepine drugs, and similar to that of sedating antidepressants. The new methodology may serve to clarify or validate premarketing abuse liability data, and may help to inform the regulatory process and physician practice. PMID- 14756710 TI - Drug driving among injecting drug users in Sydney, Australia: prevalence, risk factors and risk perceptions. AB - AIMS: To examine the prevalence of drug driving, the prevalence of drug-related motor vehicle accidents, risk perceptions of drug driving and factors associated with drug driving among injecting drug users (IDU). DESIGN: Cross-sectional survey. SETTING: Sydney, Australia. PARTICIPANTS: Three hundred current IDU. FINDINGS: Ninety-five per cent had driven a vehicle, 74% in the previous 12 months ('current drivers'). Eighty-seven per cent of life-time drivers reported having drug driven, and 88% of current drivers had drug driven in the previous 12 months. There were no significant sex differences in life-time or recent drug driving. The most common drugs used before driving in the preceding year were: cannabis, heroin, amphetamines and cocaine. A third of life-time drivers reported having had a drug driving accident, with males more likely to have done so, and 9% of current drivers reported a drug driving accident in the previous year. The most common drugs that had been used before the most recent drug driving accident were heroin, cannabis and alcohol. Alcohol was perceived to be the most dangerous substance for driving performance and cannabis the least dangerous. Recent drug drivers perceived drug driving to be less dangerous than non-drug drivers. Recent drug drivers had driven more frequently over the preceding 12 months, had significantly higher levels of dependence, higher frequency of drug use, more extensive polydrug use and were more likely to have used and/or injected a drug in a car in the previous 12 months. CONCLUSIONS: Drug driving and drug-related accidents are large-scale public health problems among IDU. These behaviours pose serious risks to IDU themselves and to the broader community. PMID- 14756711 TI - Patterns of ecstasy use in Australia: findings from a national household survey. AB - AIM: To examine the patterns, correlates and context of 'recent' (preceding 12 months) ecstasy use using data from a nationally representative sample of Australians interviewed in 2001. DESIGN: Data were analysed from the 2001 National Drug Strategy Household Survey, a multi-stage probability sample of Australians aged 14 years or older. The focus was on ecstasy use among 14-19-year olds and 20-29-year-olds, as the prevalence of recent use is highest among these groups. Recent ecstasy users were compared to those who had not used in the preceding 12 months and those who had never tried ecstasy ('others') on a range of demographic and drug use variables. Comparisons were also drawn between the patterns and context of ecstasy use of the two groups of recent ecstasy users (users aged between 14-19 and 20-29 years). FINDINGS: In 2001, 6.1% of Australians aged 14 years or older reported life-time ecstasy use, and 2.9% reported recent use. One in 10 (10.4%) of 20-29-year-olds and 5.0% of 14-19-year olds had used ecstasy recently. Although there were few demographic differences between recent users and others, compared to those who had not recently used ecstasy, recent ecstasy users were more likely to have used a range of other drugs. Although recent ecstasy users of both age groups could be characterized as polydrug users, 20-29-year-old users were more likely to use other drugs concurrently with ecstasy. CONCLUSIONS: Following cannabis and amphetamines, ecstasy is the third most widely used illicit drug in Australia. Other than a greater likelihood of having used other drugs, few demographic variables appear to distinguish recent ecstasy users from others. Australian users in their 20s use ecstasy within a context of greater polydrug use than those in their teens. Although most ecstasy users described a pattern of occasional use, minorities reported weekly use, and difficulties in reducing their use despite wishing to do so. There is a need to develop interventions to assist problematic ecstasy users to reduce their use should they wish to do so and to increase education about the potential risks of combining ecstasy with other drugs. PMID- 14756712 TI - The utility of drug testing in epidemiological research: results from a general population survey. AB - AIMS: To assess the utility of biological testing in a general population survey for estimating prevalence and evaluating self-report data quality. DESIGN: An audio computer-assisted interview was administered to subjects from June 2001 to January 2002. Immediately following the interview, subjects were requested to participate in hair, oral fluid and urine testing. SETTING: Subjects were from randomly selected households in the City of Chicago using multi-stage sampling methods. Interviews were conducted in subjects' homes. PARTICIPANTS: The data represent 627 randomly selected adult participants, ages 18-40 years. MEASUREMENTS: Prevalance, kappa, conditioned kappa, sensitivity, specificity, under-reporting, 'mixed model' and logistic regression. FINDINGS: Higher rates of marijuana use were generated from survey reports than from drug testing. Drug testing generated higher prevalence rates than survey reports for recent use of cocaine and heroin. Under-reporting of recent drug use was apparent for all three substances. Sensitivity was particularly low for cocaine and heroin. Race was related to under-reporting, with African Americans less likely to report marijuana use despite a positive test result. CONCLUSIONS: The utility of drug testing for surveys depends on the type of substance examined as well as on the type of test employed. Multiple tests have more utility than a single test. Drug testing is useful for identifying the levels and sources of under-reporting in a survey and provides a basis for adjusting prevalence estimates based on self reports. PMID- 14756713 TI - Buprenorphine in pregnant opioid-dependent women: first results of a prospective study. AB - AIM: To report results on the prospective follow-up of 34 pregnant women exposed to buprenorphine maintenance for opiate dependence. DESIGN AND SETTING: Prospective multicentre study: all pregnant women receiving buprenorphine as maintenance therapy were included as early as possible during their pregnancy. PARTICIPANTS: The pregnant women were recruited from opiate maintenance therapy centres, general practitioner-networks involved in addiction, maternity hospitals and centres for drug information during pregnancy. MEASUREMENTS: Women: drugs and medications consumed, medical and obstetrical events; offspring: withdrawal syndrome, malformation, neonatal disease. FINDINGS: The buprenorphine-exposed pregnancies resulted in 31 live births, one stillbirth, one spontaneous abortion and one voluntary termination. A neonatal withdrawal syndrome was observed in 13 cases (41.9%) and eight of these babies required opiate treatment. Two neonates had a malformation: a premature ductus arteriosus stricture and a tragus appendix. CONCLUSION: Taken together with other prospective studies, no alarming results were observed concerning pregnancy outcomes. However, further data from the comparative prospective study are required to determine whether buprenorphine can be considered as a good alternative to methadone treatment in pregnant women. PMID- 14756714 TI - Alcohol expectancies and drinking in different age groups. AB - AIMS: Because expectancies about the effects of alcohol change as drinking experience is accumulated, it is likely that the relationship of expectancy to drinking will differ with age. In this study, we examine the prediction of drinking behavior from positive and negative outcome expectancy at different ages. DESIGN: Data were collected as part of the National Alcohol Survey, using a multi-stage area probability sample of the household population of the 48 contiguous United States. PARTICIPANTS: US residents aged 12 and older (n = 2875). MEASUREMENTS: Survey questions included drinking habits (frequency, quantity, frequency of drunkenness, maximum quantity) and beliefs about the effects of alcohol (alcohol expectancies). FINDINGS: Structural equation models tested the relationship of positive and negative expectancy to drinking behavior in six age groups. Outcome expectancy accounted for a larger portion of the variance in drinking among younger respondents than among older respondents. However, suppression effects were common. When suppression effects were considered, positive expectancy predicted drinking better than negative expectancy only among respondents under 35, while negative expectancy was a better predictor of drinking status in most respondents over 35 years. Among drinkers, positive expectancy predominated over negative expectancy when suppression effects were considered. CONCLUSIONS: These results suggest that negative expectancy predicts abstention, while positive expectancy predicts level of drinking among drinkers. In expectancy research, differences between drinkers and abstainers, age of participants and the presence of suppression effects should be taken into account. PMID- 14756715 TI - Alcohol-related disorders in first- and second-generation immigrants in Sweden: a national cohort study. AB - AIMS: The risk of alcohol-related disorders in first- and second-generation immigrants in Sweden were investigated and compared with the Swedish majority population to assess how alcohol habits are modified over generations in a new society. DESIGN: Register study based on multivariate analyses of demographic data, including information on country of birth, from the Swedish Population and Housing Census of 1985 linked to data on hospital admissions for alcohol-related disorders during 1990-99 in the National Hospital Discharge Register. PARTICIPANTS: The study population consisted of a national cohort of 1.25 million youth born 1968-79 and 1.47 million adults born 1929-65. RESULTS: First- and second-generation immigrants from Finland had higher relative risks (RRs) for hospital admission because of an alcohol-related disorder compared to the Swedish majority population (socio-economic adjusted RRs 2.1 and 1.9, respectively), while first-generation immigrants born in southern Europe, the Middle East and other non-European countries had lower risks. Second-generation immigrants with heritage in southern Europe, the Middle East and other non-European countries had socio-economic adjusted RRs that were higher relative to the first generation immigrants but lower relative to the Swedish majority population. Intercountry adoptees had the highest adjusted RR (2.5). CONCLUSIONS: Patterns of alcohol abuse in the country of origin are strong determinants of alcohol-related disorders in first-generation immigrants. The patterns in second-generation immigrants are influenced by parental countries of origin as well as patterns in the majority population. The Finnish minority and intercountry adoptees are of particular concern in prevention. PMID- 14756716 TI - Heterogeneity in 12-month outcome among female and male smokers. AB - AIMS: To examine heterogeneity in outcome following treatment for smoking cessation with combined bupropion SR and behavioral counseling in women and men. DESIGN, SETTING, PARTICIPANTS: This study included 875 women and 649 men recruited from a large health-care system and randomized to one of four combinations of treatment [two dosage levels of bupropion SR (Zyban, 150 mg and 300 mg) were crossed with two counseling programs of lower and higher intensity to create a four-cell design]. MEASUREMENTS AND FINDINGS: A comprehensive set of relevant individual characteristics prior to treatment and treatment characteristics was included in the analysis. Smoking outcome at 12 months was defined as point-prevalence of any regular smoking within the 7 days prior to follow-up contact. Classification and regression tree analysis identified six subgroups in women that ranged in proportion of non-smokers from 9.8% to 42.9% and six subgroups in men that ranged in proportion of non-smokers from 17.3% to 50.0%. CONCLUSIONS: These results indicate the presence of a substantial amount of variation in treatment outcome among women and men receiving combined bupropion SR and counseling. Variation in outcome could be reduced by providing treatments tailored to subgroups of individuals who are at exceptionally high risk for smoking following cessation. PMID- 14756717 TI - Reduced dopamine D4 receptor mRNA expression in lymphocytes of long-term abstinent alcohol and heroin addicts. AB - AIM: It has been repeatedly suggested that dopamine receptor expression in peripheral blood lymphocytes reflects, to some extent, brain status. The aim of the present study was to investigate dopamine receptor expression in peripheral blood lymphocytes of long-term abstinent alcohol and heroin addicts against the background of the hypothesis, that a persisting dysfunction of the dopaminergic system contributes a biological cause to the chronic character of addiction. DESIGN: Dopamine D3 and D4 receptor mRNA expression in peripheral blood lymphocytes was measured by real-time polymerase chain reaction (PCR) in 19 alcohol addicts, abstinent for 6.2 +/- 4.7 months (mean +/- SD), and 20 heroin addicts, abstinent for 6.7 +/- 3.7 months (mean +/- SD), and compared to a control group of 29 age- and sex-matched individuals with no life-time history of substance abuse. FINDINGS: One-way anova showed significant differences in D4 mRNA expression between the groups (P = 0.005): both groups of addicts showed an approximately 50% reduction in D4 receptor mRNA expression in peripheral blood lymphocytes (PBL) compared to controls. No differences were found for D3 mRNA expression between the groups. CONCLUSION: The results of the present study indicate a withdrawal-persisting dopaminergic imbalance in abstinent addicts as measured by a suggested peripheral marker. PMID- 14756718 TI - Methadone and excessive sweating. PMID- 14756719 TI - Correcting the record: a comment on Hall et al. 2003. PMID- 14756720 TI - Errors in analyzing associations between use of smokeless tobacco and cigarettes. PMID- 14756724 TI - Female sexuality does not need a uterine cervix: no need for subtotal hysterectomy. PMID- 14756726 TI - Female genital cutting. PMID- 14756725 TI - Breech births at term revisited: new contributions from Finland and Norway. PMID- 14756727 TI - Breech birth at term: vaginal delivery or elective cesarean section? A systematic review of the literature by a Norwegian review team. PMID- 14756728 TI - Male contraception--quo vadis? AB - Recent developments in the regulation of fertility have taken place mainly within female contraception. New hormones and forms of their application are being regularly introduced. The concept of male hormonal contraception is decades old. However, until very recently, little effort has been spent trying to develop this new form of contraception for the market. Condoms and vasectomy are widely used despite their shortcomings. This reflects the willingness of male partners to share the responsibility of contraception. Condoms offer good protection from sexually transmitted disease and will therefore not be fully replaced by other forms. However, development of reliable male hormonal contraception is important in offering more choice for men and couples when choosing long-term contraception, particularly in cases where female contraception presents significant side-effects or other problems. PMID- 14756729 TI - Changes in vascular endothelial growth factor production associated with decidualization by human endometrial stromal cells in vitro. AB - BACKGROUND: The aim of this study was to clarify changes in the expression of vascular endothelial growth factor (VEGF) during decidualization by endometrial stromal cells (ESCs) in vitro. METHODS: ESCs were separated by enzymic digestion and filtration, and were cultured with RPMI 1640 in 10% fetal calf serum (FCS) and treated with medroxyprogesterone acetate (MPA) and dibutyryl-cyclic adenosine monophosphate (db-cAMP) to induce decidualization in vitro. The levels of prolactin (PRL) in the culture media were measured by enzyme immunoassay (EIA) and the levels of VEGF were measured by enzyme-linked immunosorbent assay (ELISA). The expression of VEGF mRNA by ESCs and decidualized cells was analyzed by Northern blot analysis. RESULTS: In treated cells, PRL production was significantly increased due to treatment with both db-cAMP (0.5 mmol/L) and MPA (100 nmol/L). VEGF mRNA expression was detected without any stimulation by ESCs. VEGF production was also significantly greater in cells treated with db-cAMP (0.5 mmol/L) and MPA (1 nmol/L or 100 nmol/L) than in control cells. VEGF mRNA was also increased in association with decidualization in vitro. CONCLUSIONS: VEGF production increased in association with decidualization in this in vitro study. Decidualization of ESCs may play a role in the induction of growth in the human fetus and/or placentation. The mechanism involved may include influencing the production of angiogenic growth factors such as VEGF. PMID- 14756730 TI - Reduced amount of cytochrome c oxidase subunit I messenger RNA in placentas from pregnancies complicated by preeclampsia. AB - BACKGROUND: Cytochrome c oxidase is a marker enzyme of the mitochondrial inner membrane. A change in the structure and activity of cytochrome c oxidase may alter the electron transport in the inner membrane, leading to insufficient adenosine triphosphate (ATP) production. ATP is essential for maintaining the function of cells. The aim of this study was to compare the expression of cytochrome c oxidase subunit I mRNA in placentas from normal and preeclamptic pregnancies. METHODS: By means of in situ hybridization, frozen sections of placentas from 23 women with preeclampsia and 29 women with uneventful pregnancies were examined. Digoxigenin (DIG)-labeled probes were used to detect the expression of cytochrome c oxidase subunit I mRNA in the placentas. The expression density was assessed by using an image disposal and analysis system. RESULTS: Positive expression of cytochrome c oxidase subunit I mRNA was found in the cytoplasm of villous syncytiotrophoblasts. The mean light density of cytochrome c oxidase subunit I mRNA in placental villi of normal pregnant women was 0.2638, and 0.1763 in women with preeclampsia, a statistically significant difference (P < 0.05). The number density of cytochrome c oxidase subunit I mRNA in placental villi was also significantly reduced in preeclamptic women compared with the control group (P < 0.05). CONCLUSIONS: Our study demonstrates a reduced amount of cytochrome c oxidase subunit I mRNA in preeclamptic placentas compared to control placentas. We hypothesize that a reduced expression may play a role in the pathophysiology of preeclampsia. PMID- 14756731 TI - Ultrasound estimates of gestational age among perinatally demised: a population based study. AB - BACKGROUND: Correct estimation of gestational age may improve the quality of obstetric care. We hypothesize that significant differences between traditional and alternative estimates by ultrasound are evident among perinatal deaths. METHODS: Population-based case series with data linkage between autopsy records and The Medical Birth Registry of Norway, including perinatally demised singletons who were examined by autopsy and post-mortem radiography, having antenatal estimates of gestational age both by the calendar method, as calculated from the first day of the last menstrual period preceding the pregnancy (GAlmp), and by mid-second-trimester ultrasound (GAus), N = 380. The main outcome measure was the distribution of GAlmp and GAus within weight strata. RESULTS: Mean GAus was 1 week less than mean GAlmp (t-test, p < 0.001). The degree of apparent growth restriction manifest after death, as expressed by both birthweight and by post-mortem radiographic measurements, was fairly well correlated with the degree of downward adjustment of age by ultrasound in the early second trimester (Pearson's correlation, r = - 0.599, p < 0.001). The degree of discrepancy between the ultrasound and the calendar methods was associated both with placenta findings (Kruskal-Wallis test, chi2 = 20.95, p = 0.007) and with the main cause of death (Kruskal-Wallis test, chi2 = 27.65, p = 0.004). CONCLUSION: Among infants who died perinatally, gestational age seemed to be systematically downward adjusted by mid-second-trimester screening ultrasound, particularly among those who were the most growth retarded at the time of death. PMID- 14756732 TI - Plasma homocysteine in early and late pregnancies complicated with preeclampsia and isolated intrauterine growth restriction. AB - BACKGROUND: Elevated circulating homocysteine is an independent risk factor for cardiovascular disease. Increased homocysteine plasma levels have been reported to occur in approximately 20-30% of women with preeclampsia and it has been suggested that they may predict the subsequent development of preeclampsia. METHODS: In a cohort of 1874 pregnant women followed longitudinally, who participated in the Down screening program, 27 developed preeclampsia and 36 intrauterine growth restriction (IUGR). A control group of 63 uneventful pregnancies was selected. Plasma homocysteine was assayed in the early second trimester and at delivery in all groups. Data were compared with Wilcoxon's matched-pair test. RESULTS: No statistically significant difference of plasma homocysteine between controls and preeclamptic or IUGR pregnancies in the early second trimester were found. There was a significant difference, only at delivery, between the preeclamptic subjects and the controls. CONCLUSIONS: We failed to demonstrate a plasma homocysteine predictive value in pregnancies subsequently complicated by preeclampsia and IUGR. As previously stated, we found that an elevated homocysteine plasma level is associated with overt preeclampsia. PMID- 14756733 TI - Arterial oxygen tension during sleep in the third trimester of pregnancy. AB - BACKGROUND: Changes during pregnancy include reduced functional residual capacity (FRC) and residual volume (RV), increased alveolar difference for oxygen and, in the supine position, reduced cardiac output. In conjunction with sleep-related disturbances, these changes could lead to maternal oxygen desaturation during sleep. OBJECTIVES: Because of conflicting data from respiratory sleep studies in pregnancy, we performed complete polysomnography on 21 pregnant women at the 36th week of gestation and again postpartum. We also measured the partial pressure of oxygen in the arterial blood (PaO2) in the supine and sitting positions. METHODS: We tested 21 healthy pregnant women at the 36th week of gestation. Arterial samples were taken in the sitting position. Complete polysomnography was performed in all of the pregnant women. Before the polysomnography arterial samples were taken in the supine and sitting positions and then every 2 h until termination of the study. RESULTS: We did not find any correlation between SaO2/PaO2 levels and apnea, hypopnea or percent of rapid eye movement (REM) sleep. The frequency of apnea and hypopnea was significantly lower during pregnancy (5.81 +/- 2.1 apneas or hypopneas per hour of sleep) than postpartum (12.1 +/- 2.7 apneas or hypopneas per hour of sleep) (p < 0.001), which may be due to the raised level of progesterone. The PaO2 levels in the supine position were significantly lower than in the sitting position at 36 weeks of gestation (p < 0.001). No differences were found between PaO2 levels in the sitting and supine positions postpartum (p < 0.5). CONCLUSIONS: According to our results we conclude that 1) the frequency of apnea and hypopnea in pregnancy was significantly lower than postpartum, and 2) a significant difference in PaO2 levels in the sitting and supine positions was observed at 36 weeks of gestation. PMID- 14756735 TI - Human chorionic gonadotropin in maternal serum in relation to fetal gender and utero-placental blood flow. AB - BACKGROUND: To investigate whether fetal gender differences in human chorionic gonadotropin (hCG) in maternal serum and the presence of hCG receptors in the wall of the uterine arteries influence the utero-placental blood flow. METHOD AND MATERIAL: Sixty-six healthy women with singleton uncomplicated pregnancies were examined at 8-10, 16-19 and 31-37 weeks of gestation. The pulsatility index (PI) was measured in the uterine arteries, simultaneously with sampling of peripheral maternal blood for hCG determination. Volume flow in the uterine arteries was determined in the second and third trimesters only. RESULTS: In the first and second trimesters no gender differences in the hCG levels were observed. From the second to the third trimester the hCG levels increased significantly in pregnancies with female fetuses (P < 0.05), while in pregnancies with male fetuses the hCG levels tended to decline. The PI declined significantly from the first to the third trimester in both genders (P < 0.001). In the first and third trimesters no gender differences were seen. In the second trimester the PI values were significantly higher in pregnancies with male fetuses than in those with female fetuses (P < 0.02). The flow volume increased significantly in both genders from the second to the third trimester (P < 0.001). In the third trimester the flow volume was higher in pregnancies with female fetuses than in those with male fetuses (P = 0.05). CONCLUSION: The gender differences in uterine artery PI and flow volume were not correlated to maternal serum hCG levels. PMID- 14756734 TI - Correlations between umbilical and maternal serum adiponectin levels and neonatal birthweights. AB - OBJECTIVE: To measure adiponectin levels in maternal serum and umbilical cord serum at delivery, and examine whether or not there are correlations between adiponectin levels and neonatal birthweights, maternal body weights and body mass indexes. STUDY DESIGN: The study included 84 healthy mothers who had given birth to healthy neonates. Adiponectin levels in maternal serum and umbilical cord serum were determined by radioimmunoassay and compared. RESULTS: The ranges of adiponectin levels for umbilical cord serum and maternal serum were 22.7-78.4 microg/ml and 4.0-43.3 microg/ml, respectively. Umbilical serum adiponectin levels (46.9 +/- 1.2 microg/ml) were significantly higher than maternal serum adiponectin levels (16.1 +/- 0.8 micro g/ml) (p < 0.001). No correlation was found between the adiponectin levels in maternal serum and those in umbilical cord serum (r = 0.158, p = 0.151). Umbilical serum adiponectin levels were significantly correlated with both neonatal birthweights (r = 0.454, p < 0.001) and gestational ages at birth (r = 0.295, p = 0.006), but not with maternal serum adiponectin levels. Maternal serum adiponectin levels were only negatively correlated to maternal weights and body mass index at delivery (r = 0.288, p = 0.008; r = 0.372, p < 0.001). CONCLUSION: The levels of adiponectin were higher in umbilical cord serum than in maternal serum. Moreover, the adiponectin levels in umbilical cord serum were found to correlate positively with neonatal birthweights. Therefore, fetal adiponectin, not maternal serum adiponectin, may be involved in fetal development during late pregnancy. PMID- 14756736 TI - Treatment of postpartum thrombotic microangiopathy with plasma exchange using cryosupernatant as replacement. AB - Hemolytic uremic syndrome (HUS) and thrombotic thrombocytopenic purpura (TTP) are rare but acknowledged problems of pregnancy and the postpartum period. These diseases together with thrombotic angiopathy are associated with high maternal and fetal mortality and severe long-term morbidity. We describe four women with postpartum HUS/TTP treated with plasma exchange cryosupernatant fraction of plasma (CFP) as replacement fluid. Anuria or oliguria at the beginning of treatment and thrombocytopenia [thromb- (29-68) x 109/L] were common features. Two of the patients developed a prolonged and more difficult clinical condition affecting the central nervous system and the liver and their platelet counts remained low despite the plasma exchange. The renal and hepatic function of all of the patients recovered fully. This small analysis lends weight to early plasma exchange with cryosupernatant as part of the treatment of postpartum HUS/TTP and suggests that persistent thrombocytopenia is a signal of more serious disease. PMID- 14756737 TI - Are health expectations of term breech infants unrealistically high? AB - BACKGROUND: The aim of this study was to compare the effect of fetal presentation and mode of delivery on infant outcome in a nation-wide study. METHODS: In a retrospective observational cohort study, we compared, with the help of Finnish Medical Birth Register and other nation-wide registers, the short-term and long term outcome of infants born by breech vaginal (n = 1270) or by vertex vaginal delivery (n = 128,683) or through planned cesarean section (CS) in breech (n = 1640) or vertex (n = 4997); the pregnancies were otherwise entirely normal. RESULTS: One perinatal death occurred in the breech vaginal group and 23 deaths in the vertex vaginal group (p = 0.112), but none in either CS group. Breech vaginal delivery was associated with increased risk of Apgar scores 6 or less at age 1 min (OR 7.65, CI 6.41-9.12) and at age 5 min (OR 6.42, CI 4.36-9.45) as compared with vertex vaginal delivery. These odd ratios were also elevated (OR 4.59, CI 3.48-7.08 and OR 7.58, CI 3.09-18.66, respectively) when compared with breech planned CS. Yet the risk for birth trauma of infants in the breech vaginal group was smaller (OR 0.70, CI 0.51-0.96) than that in the vertex vaginal group but this risk was smallest in the planned CS groups. A number of other neonatal complications occurred equally commonly in each group. Breech infants born vaginally needed fewer admissions (OR 0.58, 0.47-0.72) to out-patient departments and the cumulative incidence of long-term morbidity in the breech vaginal group was smaller (OR 0.47, CI 0.28-0.80) to the age of 7 years than that in the breech planned CS. The maturity for starting school and school performance during the first two school years showed no dependence on mode of delivery. CONCLUSION: Apart from Apgar suppression, elective vaginal delivery of a full-term breech fetus in highly selected pregnancies does not cause additional neonatal hazards as compared with full-term vertex deliveries. The immediate outcome was best for breech or vertex infants born through elective CSs. PMID- 14756738 TI - Prevalence of female genital mutilation among African women resident in the Swedish county of Ostergotland. AB - OBJECTIVES: To establish the prevalence of female genital mutilation (FGM) among African women resident in the Swedish County of Ostergotland and assess the types of FGM. MATERIAL AND METHODS: Three hundred and four African women aged > or = 18 years were domiciled in Ostergotland by the end of 1998. The women were invited by letter. A socio-cultural questionnaire designed to give an overall picture of FGM within a socio-economic context, and also to invite the women to an interview and examination, was sent to all African women in the county of Ostergotland. Women who gave their consent (n = 63) underwent a gynecologic examination. RESULTS: The response rate was 84%. According to the questionnaire, 68% of all the African women were genitally mutilated. The clinical examination revealed that 39 women (62%) were mutilated, 17 of them (44%) had undergone removal of part or all external genitalia and stitching ('infibulation'), 26% had undergone removal of the prepuce of the clitoris ('prepucectomy'), 23% had undergone various cultural practices on the external genitalia, and 7.7% excision of the clitoris with partial or total removal of labia minora ('clitoridectomy'). CONCLUSION: The influx of immigrants to Sweden and the other Scandinavian countries from cultures where FGM is practiced, requires that physicians and other health professionals familiarize themselves with the practice and the cultural beliefs underlying it. Sensitivity to the needs of these women as well as attention to the potential physical hazards posed by the practice are important factors in care. PMID- 14756739 TI - Sexuality after total vs. subtotal hysterectomy. AB - BACKGROUND: The effect of hysterectomy on sexuality is not fully elucidated and until recently total and subtotal hysterectomies have only been compared in observational studies. AIMS: To compare total abdominal hysterectomy (TAH) to subtotal abdominal hysterectomy (SAH) regarding effects on sexuality. METHODS: In a Danish multicenter trial 319 women were randomized to TAH (n = 158) or SAH (n = 161); 185 women had self-selected TAH (n = 80) or SAH (n = 105) in a simultaneously conducted observational study. Women were followed for 1 year by strict data collection procedures, including postal questionnaires. Results were analyzed by intention to treat (ITT) analyses. RESULTS: No significant differences were observed between TAH and SAH at 1-year follow-up in both the randomized trial and the observational study regarding women's desire for sex, frequency of intercourse, frequency of orgasm, quality of orgasm, localization of orgasm, satisfaction with sexual life, and dyspareunia. None of these sexual variables changed significantly from entry to the 1-year follow-up, apart from dyspareunia, which was significantly (p = 0.009) reduced in both intervention groups. Significant (p < 0.05) predictors for satisfaction with sexual life after hysterectomy were the preoperative satisfaction with sexual life [odds ratio (OR) 32, 95% confidence interval (CI) 10-125], good relationship with partner (OR 50, 95% CI 9-354), physical well-being (OR 0.30, 95% CI 0.09-0.88) and hormone replacement therapy (OR 0.23, 95% CI 0.06-0.78). CONCLUSIONS: Both TAH and SAH significantly reduce dyspareunia without having a negative effect on sexual function. The shift toward SAH seems unwarranted. PMID- 14756740 TI - Menstrual bleeding patterns in pre- and perimenopausal women: a population-based prospective diary study. AB - BACKGROUND: To investigate natural menstrual bleeding patterns in pre- and perimenopausal women, a prospective observational population study was carried out. METHODS: A total of 1616 (80.8%) of a population-based sample of 2000 Danish women aged 45-54 years answered an initial questionnaire. Of 1059 women with natural gynecological functions, 951 (89.8%) completed 1 year of daily recording of bleeding. Of these women, 592 were pre- or perimenopausal and were included in this study. MAIN OUTCOME MEASURES: Length and heaviness of bleeding episodes, including frequency of spotting, flooding and prolonged bleeding, according to regularity of cycles. RESULTS: The proportion of women with irregular cycles increased from 58.3% at age 45-46 years to 100% at age 53-54 years (P < 0.001). Irregularity of cycles was accompanied by increased variation in both length and subjective assessment of heaviness of bleeding episodes in the individual woman and between women (P < 0.001). Episodes of spotting (P < 0.001) and prolonged bleeding (10 days or more) (P < 0.001) were more common in women with irregular cycles. Subjective assessment of heaviness of bleeding episodes, however, was higher in women with regular cycles (P < 0.001). One or more episodes of flooding were recorded by 24.0% of all women irrespective of regularity of cycles (P = 0.40) but more frequently in younger women (P < 0.001). CONCLUSIONS: Irregularity of cycle lengths at the end of the fertile period is common, and is accompanied by an increased inter- and intraindividual variation in the length and heaviness of bleeding episodes, including increased frequency of prolonged bleeding and spotting. PMID- 14756741 TI - Frequency of spontaneously occurring postmenopausal bleeding in the general population. AB - BACKGROUND: To determine the frequency of spontaneously occurring postmenopausal bleeding in the population, a prospective observational population study was undertaken. METHODS: Out of a population-based sample of 2000 Danish women aged 45-54 years, 1616 women responded to an initial questionnaire. Of these, 1059 women with natural gynecologic functions were eligible for the diary study, and of these, 951 (89.8%) women completed 1 year of daily recording of bleeding, medication relevant to bleeding, and medical consultations for gynecologic matters. Of the 951 women, 271 were postmenopausal. MAIN OUTCOME MEASURES: Occurrence of spontaneous postmenopausal bleeding including strength and length of each bleeding. RESULTS: Of 271 postmenopausal women, 29 (10.7%, 95% confidence interval 7.3-15.0%) recorded spontaneous postmenopausal bleeding. The incidence of women with bleeding was strongly correlated with time since menopause (P = 0.0001), with an estimated incidence of 409/1000 (218/1000-771/1000) person-years immediately after the first 12 months of amenorrhea following the menopause falling to 42/1000 (17/1000-102/1000) more than 3 years after menopause. CONCLUSIONS: This study indicates that recurrence of bleeding is very common in the time immediately after the 12 months of amenorrhea after what is currently defined as the final menstrual period, declining to low frequencies more than 3 years after the final menstrual period. PMID- 14756742 TI - Ovarian metastasis in women with clinical stage I endometrial carcinoma. AB - BACKGROUND: The incidence of ovarian metastasis in women with clinical stage I endometrial carcinoma is generally reported to be 5%, leading to the practice of removing the ovaries at surgery even in young patients. METHODS: A retrospective study of 84 patients with clinical stage I endometrial cancer was carried out. Patients were excluded if the pathologic study revealed any evidence of extrauterine, apart from adnexal, spread or if the peritoneal cytology was positive. Patients with serous papillary or clear cell tumor histology were also excluded. RESULTS: Sixty-seven patients fulfilled the inclusion criteria. Only three (4%) patients were found to be in surgical stage IIIA, all three had grade 3 tumors. Of these patients, two had uterine serosal involvement and one had a microscopic tumor implant in a fallopian tube; none had ovarian metastasis. CONCLUSIONS: The risk of ovarian metastasis in women with well to moderately differentiated endometrial cancer, myometrial invasion limited to less than one half of the myometrium, negative peritoneal cytology and no evidence of metastatic lymph node spread is negligible. Young patients with a preoperative histological diagnosis of well to moderately differentiated endometrial carcinoma may be surgically staged, leaving the final decision regarding removal of the ovaries pending a thorough pathological review of the surgical specimens. PMID- 14756743 TI - Advanced cervical pregnancy: uterus-sparing therapy initiated with a combination of methotrexate and mifepristone followed by evacuation and local hemostatic measures. PMID- 14756744 TI - Familial recurrent molar pregnancy: a case report. PMID- 14756745 TI - Malacoplakia of the cervix uteri and vulva. PMID- 14756746 TI - Possible causes of low sensitivity of HPV-DNA testing for CIN2/3. PMID- 14756748 TI - Effect of caudal epidural xylazine on intraoperative distress and post-operative pain in Holstein heifers. AB - OBJECTIVE: To compare the effects of caudal epidural xylazine versus saline on tolerance of paravertebral nerve block and flank surgery and on post-operative pain in heifers used for a veterinary student training laboratory. STUDY DESIGN: Randomized controlled prospective study. ANIMALS: Fourteen one-year-old, nongravid, healthy Holstein heifers, weighing 360 +/- 5 kg. METHODS: Xylazine (0.05 mg kg(-1)) or 0.9% saline (5 mL) was injected using a caudal epidural technique to seven heifers undergoing a flank surgery. Nerve block of the right paravertebral fossa was performed using equal parts of lidocaine 2% and bupivacaine 0.5%. Heart and respiratory rates, rectal temperature, rumination frequency, and appetite were recorded before and at 4, 8, and 24 hours after surgery. Scores were recorded for: tolerance of local anesthesia injections (pre operatively), sedation, ataxia and distress (intraoperatively, every 30 minutes), and pain (4, 8, and 24 hours post-operatively). RESULTS: The animals reaction to local anesthetic injection was judged to be less in the xylazine group by both an experienced observer (p<0.001) and student surgeons (p<0.01). The xylazine group required less local anesthetic (82.9 +/- 13.8 mL) versus the saline group (108.4 +/- 19.6 mL, p=0.035). Intraoperatively, xylazine heifers were more sedated at all times (p-values from <0.001 to 0.017), were more ataxic for the first 1.5 hours (p-values from <0.001 to 0.026), and lower in distress at all times (p values from <0.001 to 0.007). No difference in post-operative pain or physiologic variables was found, except immediately post-operatively, rectal temperature was higher in the xylazine group (39.5 +/- 0.3 degrees C) than in the saline group (38.6 +/- 0.2 degrees C, p<0.001). CONCLUSION AND CLINICAL RELEVANCE: Compared with epidural saline, caudal epidural xylazine reduced distress of anesthetic injection and surgical manipulation in heifers and an improvement in animal well being was apparent. This effect may have been as a result of sedation. Pre operative epidural xylazine did not appear to improve post-surgical analgesia in our study. PMID- 14756749 TI - Cardiopulmonary effects of combined xylazine-guaiphenesin-ketamine infusion and extradural (inter-coccygeal lidocaine) anaesthesia in calves. AB - OBJECTIVE: To investigate the cardiopulmonary effects of a xylazine-guaiphenesin ketamine infusion combined with inter-coccygeal extradural (lidocaine) anaesthesia in calves. STUDY DESIGN: Prospective study. ANIMALS: Five Holstein Friesian calves (one steer, four heifers) aged 6 weeks weighing 65.2 +/- 2.7 kg. MATERIALS AND METHODS: Calves were anaesthetized with isoflurane in oxygen for instrumentation. At least 12 hours later, xylazine (0.2 mg kg(-1) i.m.) was given. After 15 minutes, an infusion of xylazine hydrochloride (0.1 mg mL(-1)), guaiphenesin (50 mg mL(-1)) and ketamine (1 mg mL(-1)) (X-G-K) was infused at a rate of 1.1 mL kg(-1) hour-1 i.v. Oxygen (4 L minute(-1)) was delivered by nasotracheal tube 30 minutes later. Inter-coccygeal (Co1-Co2) extradural anaesthesia (lidocaine 2%, 0.18 mL kg(-1)) was administered 30 minutes later. Cardiopulmonary variables were obtained in the unsedated standing calves 10 minutes after xylazine, 15 and 30 minutes after X-G-K without O2, 15 and 30 minutes after X-G-K with O2 and 5, 15, 30, 45 and 60 minutes after extradural anaesthesia. Data were analysed using a repeated measurement analysis of variance including an autoregressive covariance structure of order 1 (correlations at different time intervals). RESULTS: Xylazine caused significant (p<0.05) decreases in heart rate (HR), cardiac output (Qt) and index (CI), stroke volume and stroke index, mean, systolic and diastolic arterial blood pressure (MAP, SAP, DAP), left (LVWSI) and right ventricular stroke work index (RVWSI), mean, systolic and diastolic pulmonary arterial pressure (MPAP, SPAP, DPAP), arterial pH, arterial oxygen tension (PaO2), arterial base excess, arterial HCO3- concentration, arterial saturation, packed cell volume, arterial and venous oxygen content (CaO2, CvO2), O2 consumption and O2 delivery (VO2, DO2). Increases in systemic vascular resistance (SVR) and pulmonary vascular resistance (PVR) were observed. During X-G-K infusion without O2, HR, Qt and CI increased gradually while SVR, PVR and MAP decreased. Left ventricular stroke work index and PaO2 remained constant, while O2 supplementation improved PaO2. Coccygeal extradural anaesthesia had little effect on cardiopulmonary variables. Respiratory rate (f) and PaCO2 significantly increased over the experiment. CONCLUSIONS AND CLINICAL RELEVANCE: Xylazine caused adverse cardiopulmonary effects in calves. Improvement occurred during xylazine-guaiphenesin-ketamine infusion. Cardiac index and arterial blood pressure remained below baseline values while sustained increases in respiration rate and PaCO2 were observed. Inter-coccygeal extradural anaesthesia had only minor effects. Oxygen supplementation proved advantageous during guaiphenesin, ketamine and xylazine infusion in healthy calves in combination with coccygeal extradural anaesthesia induced persistent cardiopulmonary depression. PMID- 14756750 TI - Intra-articular lidocaine plus bupivacaine in sheep undergoing stifle arthrotomy. AB - OBJECTIVE: To evaluate the effect of intra-articular (i.a.) lidocaine plus bupivacaine on post-operative pain in sheep undergoing stifle arthrotomy. STUDY DESIGN: Randomized controlled experimental trial. ANIMALS: Sixteen adult Rambouillet-cross ewes. METHODS: Sheep were randomly assigned to one of two treatment groups. The lidocaine/bupivacaine group (L/B, n=8) received i.a. lidocaine (40 mg (2 mL)) prior to incision and i.a. bupivacaine (10 mg (2 mL)) post-closure, while the control group (n=8) received no i.a. injections. i.a. local anesthetics were an addition to the standard analgesic protocol of phenylbutazone (1 g orally, every 24 hours for 5 days) and transdermal fentanyl (equivalent to 15 mg), initiated 24 hours prior to surgery. A stifle arthrotomy was performed with the purpose of creating a full-thickness articular cartilage defect. Two observers blinded to treatment assessed sheep for total pain score using a numeric ranking scale that included: comfort, movement, and flock behavior. The first observation (T=0) was obtained the evening of surgery (3-7 hours post-operatively); subsequent observations occurred every 12 hours for 72 hours. Nonparametric statistical tests were used to evaluate differences between groups for total pain score. RESULTS: L/B sheep had significantly lower total pain scores at T=0 than control sheep (p<0.05). No significant differences between treatments were noted at any subsequent time periods. There were no differences attributable to the use of different observers. CONCLUSIONS AND CLINICAL RELEVANCE: i.a. lidocaine plus bupivacaine provided analgesia at 3-7 hours post-operatively. Use of i.a. lidocaine and bupivacaine is a simple, effective, yet inexpensive perioperative analgesic protocol for joint surgery in sheep. PMID- 14756751 TI - Evaluation of a new oscillometric blood pressure monitor in isoflurane anesthetized dogs. AB - OBJECTIVE: This study was conducted to evaluate the performance of a new veterinary oscillometric noninvasive blood pressure (NIBP) monitor in anesthetized dogs. STUDY DESIGN: Assessment was made to determine how closely indirect measurements were associated with direct measurements, and if there were statistically significant differences between the measurements by site. ANIMALS: Six mongrel dogs weighing 27.8 +/- 2.9 kg. METHODS: Dogs were anesthetized with thiopental and maintained with isoflurane, which was delivered with controlled ventilation. Direct pressure measurements were obtained via a percutaneously placed arterial catheter. A range of systolic arterial pressures (SAP) were achieved by changing the isoflurane concentrations. Sites of cuff placement for indirect measurements were identified as metacarpus, metatarsus, and anterior tibial. RESULTS: At pressures below 80 mm Hg, indirect systolic measurements averaged 4 +/- 3 mm Hg, higher than the direct values. At normal and high levels, indirect systolic measurements underestimated direct values by 18 +/- 6 and 23 +/ 6 mm Hg, respectively. Diastolic and mean pressure measurements followed the same trend, with indirect values being lower than the direct arterial pressures. Systolic, diastolic and mean arterial pressure measurements differed by cuff placement site. CONCLUSIONS: When analyzed by site and level, indirect systolic and mean arterial blood pressures during hypotension were essentially the same as direct pressures. However, at pressures within the normal or high range, indirect measurements underestimated the direct pressures. CLINICAL RELEVANCE: Noninvasive blood pressure measurements with a new oscillometric monitor provided an excellent means of detecting arterial hypotension in anesthetized dogs. The metatarsal site for cuff placement was slightly better than the metacarpal or anterior tibial site, considering that the regression line was closest to complete equality between the indirect and direct measurements for SAP. PMID- 14756752 TI - Effects of acepromazine on the incidence of vomiting associated with opioid administration in dogs. AB - OBJECTIVE: To evaluate the anti-emetic properties of acepromazine in dogs receiving opioids as pre-anesthetic medication. STUDY DESIGN: Randomized prospective clinical study. ANIMALS: One hundred and sixteen dogs (ASA I or II), admitted for elective surgical procedures. The dogs were a mixed population of males and females, purebreds and mixed breeds, 0.25-13.4 years of age, weighing 1.8-57.7 kg. METHODS: A prospective clinical trial in which the dogs were randomly assigned to one of three groups. All groups received acepromazine (0.05 mg kg(-1) intramuscularly (i.m.)). Group I received acepromazine 15 minutes prior to opioid administration. Group II received acepromazine in combination with the opioid. Group III received acepromazine 15 minutes after opioid administration. One of three different opioids was administered i.m. to each dog: morphine sulfate at 0.5 mg kg(-1); hydromorphone hydrochloride at 0.1 mg kg(-1); or oxymorphone hydrochloride at 0.075 mg kg(-1). RESULTS: Dogs receiving acepromazine before the opioid (group I) had a significantly lower incidence of vomiting (18%) than dogs in groups II (45%) and III (55%). The degree of sedation was significantly lower in the dogs receiving the combination of acepromazine and the opioid (group II) than in dogs receiving the opioid as the first drug (group III). CONCLUSIONS AND CLINICAL RELEVANCE: Acepromazine administered 15 minutes before the opioid lowers the incidence of vomiting induced by opioids. PMID- 14756753 TI - Evaluation of intraperitoneal and incisional lidocaine or bupivacaine for analgesia following ovariohysterectomy in the dog. AB - OBJECTIVE: To determine if intraperitoneal (i.p.) and incisional (s.c.) lidocaine or bupivacaine provide analgesia following ovariohysterectomy (OHE). STUDY DESIGN: Prospective, randomized, controlled, blinded clinical trial. ANIMALS: Thirty dogs presenting to the Veterinary Teaching Hospital for elective OHE. METHODS: Dogs were pre-medicated with acepromazine and butorphanol, induced with thiopental and maintained with isoflurane. They were randomly assigned to three groups: 10 received 8.8 mg kg(-1) 2% lidocaine with epinephrine i.p. (LID); 10 received 4.4 mg kg(-1) 0.75% bupivacaine i.p. (BUP); and 10 received 0.9% saline i.p. (SAL) upon completion of OHE. All i.p. doses were standardized to 0.88 mL kg(-1) with saline. An additional 2 mL of undiluted solution was placed s.c. prior to incisional closure. Dogs were scored at 0.5, 1, 2, 3, 6, 8 and 18 hours post-extubation by one observer. Dogs were evaluated using a visual analogue scale (VAS) for pain and sedation, and a composite pain scale (CPS) that included physiologic and behavioral variables. Dogs were treated with 0.22 mg kg(-1) butorphanol + acepromazine if their VAS (pain) score was >50. Parametric variables were analyzed using Student's t-test or repeated measures ANOVA as appropriate. Non-parametric variables were analyzed by chi2-test. RESULTS: There were no significant differences in age, weight, incision length, surgery time, anesthesia time, or total thiopental dose among groups. Peak post-surgical pain scores for all groups occurred at 0.5 hours and returned to baseline by 18 hours. Dogs in the BUP group had significantly lower VAS-pain scores overall than dogs in the SAL group. Seven out of 10 dogs in the SAL group, 4/10 in the LID group and 2/10 in the BUP group were treated with supplemental acepromazine and butorphanol. No differences between groups were detected with the CPS. No adverse side-effects were observed. CONCLUSIONS AND CLINICAL RELEVANCE: Our findings support the use of i.p. and s.c. bupivacaine for post-operative analgesia following OHE in the dog. PMID- 14756754 TI - Systemic lidocaine infusion as an analgesic for intraocular surgery in dogs: a pilot study. AB - OBJECTIVE: To determine if systemic administration of lidocaine during intraocular surgery reduces post-operative ocular pain. STUDY DESIGN: Randomized, masked, controlled experimental trial. ANIMALS: Twelve dogs weighing 15.5 +/- 1.7 kg (mean +/- SD) and aged 2.5 +/- 0.6 years. METHODS: All dogs underwent a baseline ophthalmic examination and subjective pain score. Anesthesia consisted of acepromazine (0.1 mg kg(-1), i.m.), propofol (4-6 mg kg(-1), i.v.), and isoflurane in oxygen. There were three groups each receiving a bolus followed by an infusion (n=4): saline (0.3 mL kg(-1) i.v. + 0.2 mL kg(-1) hour(-1) i.v.); morphine (0.15 mg kg(-1) i.v. + 0.1 mg kg(-1) hour(-1) i.v.); and lidocaine (1.0 mg kg(-1) i.v. + 0.025 mg kg(-1) minute(-1) i.v.). All treatments began 15 minutes prior to starting of phacoemulsification and lens removal from the right eye. Pain scores were recorded at 0.5, 1, 2, 3, 4, 6, 8, 16, and 24 hours after t=0 (extubation). Rescue morphine was administered (1.0 mg kg(-1) i.m.) if the subjective pain score > or =9 (maximum=24), and the dog was excluded from further data analysis. Differences in pain scores and time-to-treatment failure (TTF) were analyzed using the Wilcoxon's rank sum test. Differences in incidence of treatment failure were analyzed using Fisher's exact test. Physiologic data were analyzed using repeated measures ANOVA. Significance was defined as P<0.05. RESULTS: Incidence of treatment failure was 100% in saline-treated dogs and 50% in morphine- or lidocaine-treated dogs. There was no difference in intraocular pressure, aqueous flare, cell count (or protein) between groups in the operated eye at any time following extubation. CONCLUSION AND CLINICAL RELEVANCE: This pilot study suggests that intraoperative lidocaine may provide analgesic benefits similar to morphine for intraocular surgery in dogs, but more definitive research is needed. This model appears to be appropriate for pain assessment studies as the negative control group demonstrated 100% failure rate. PMID- 14756755 TI - The cardiovascular dose-response effects of isoflurane alone and combined with butorphanol in the green iguana (Iguana iguana). AB - OBJECTIVE: To assess the cardiovascular effects (arterial blood pressure, heart rate, and metabolic acid-base status) of three doses (MAC multiples) of isoflurane alone and combined with butorphanol in the green iguana (Iguana iguana). STUDY DESIGN: Prospective randomized double-blind, two-period cross-over trial. ANIMALS: Six mature healthy green iguanas (Iguana iguana). METHODS: The iguanas received each of two treatments, saline 0.1 mL kg(-1) (SAL) and butorphanol 1.0 mg kg(-1) (BUT) during isoflurane anesthesia. Treatments were separated by at least 1 week. The iguanas were exposed to each of the three minimum alveolar concentration (MAC) multiples (1.0, 1.5, and 2.0) in random order. Anesthesia was induced with isoflurane and maintained using controlled ventilation. Instrumentation included use of an ECG, airway gas monitor, cloacal thermometer, esophageal pulse oximeter, and the placement of a femoral arterial catheter. Body temperature was stabilized and maintained at 32 degrees C. The treatment was administered, and the animals were equilibrated for 20 minutes at each MAC multiple. At each concentration, the heart rate, blood pressure (systolic, mean, diastolic), end-tidal CO2, and SpO2 were measured. At 1.0 and 2.0 MAC, simultaneous blood samples were drawn from the tail vein/artery complex and femoral catheter for blood gas analysis. Data were analyzed using a two-way analysis of variance for repeated measures looking for differences between treatments and among MAC multiples. RESULTS: There were no significant differences in any of the cardiovascular variables between the treatments. Significant differences among isoflurane MAC multiples were observed for HR, mean, diastolic, and systolic blood pressures. Blood pressure and heart rate decreased with an increasing dose of anesthetic. There were no significant differences between treatments or MAC multiples for any of the blood gas variables. The blood pH, PCO2, HCO3-, and hemoglobin saturation differed significantly between sites. Pulse oximetry values measured from the carotid complex did not correlate with and were significantly different from the calculated hemoglobin saturation values determined using the gas analyzer. CONCLUSION AND CLINICAL RELEVANCE: Cardiovascular depression associated with isoflurane anesthesia in the green iguana is dose dependent. The degree of cardiovascular depression was not significantly different when isoflurane was combined with butorphanol. This finding suggests that the pre-emptive or intraoperative use of butorphanol is unlikely to be detrimental to cardiovascular function. Butorphanol may be a useful anesthetic adjunct to isoflurane anesthesia in the green iguana. PMID- 14756756 TI - Effects of levomepromazine and different desflurane concentrations upon electrocardiographic variables in dogs. AB - OBJECTIVES: To investigate the effects of levomepromazine and different desflurane concentrations upon electrocardiographic variables. ANIMALS: Twenty adult mongrel dogs of both sexes weighing 6-28 kg. METHODS: Dogs were divided into two groups of 10 animals. Group 1 received 1 mg kg(-1) i.v. of levomepromazine and 15 minutes later anesthesia was induced with propofol (3 mg kg(-1) i.v.). Desflurane end-tidal concentration was set at 1.6 MAC. After 30 minutes at this concentration, measurements were taken and the end-tidal concentration was reduced to 1.4 MAC. Thereafter, it was reduced to 1.2 and then 1.0 MAC at 15-minute intervals. The same procedure was followed for group 2, except that levomepromazine was replaced with 0.2 mL kg(-1) of 0.9% saline solution and more propofol was needed for induction (7 mg kg(-1)). The animals' body temperature was maintained between 38.3 and 39 degrees C using a heating pad. The electrocardiographic tracing was obtained from lead II throughout the experimental period. The measurements were taken immediately before the administration of levomepromazine or placebo (T1), 15 minutes after pre medication (T2) and 30 minutes after the establishment of 1.6 MAC (T3). The other measurements were made at the concentrations of 1.4, 1.2, and 1.0 MAC, respectively (T(4-6)). The numerical data were submitted to analysis of variance plus F-test (p<0.05). RESULTS: The dogs that received levomepromazine had a decrease in heart rate. However, in both groups it increased with desflurane administration. Levomepromazine, in association with desflurane, did not induce significant electrocardiographic changes, and all mean values (except P-wave duration) were within the reference range for this species. CONCLUSIONS AND CLINICAL RELEVANCE: This study documented that levomepromazine, in association with desflurane, does not induce significant changes in electrocardiographic variables, suggesting that this drug combination has minimal effect on myocardial conduction. PMID- 14756757 TI - ydk1-D, an auxin-responsive GH3 mutant that is involved in hypocotyl and root elongation. AB - To study the GH3 gene family of Arabidopsis, we investigated a flanking sequence database of Arabidopsis activation-tagged lines. We found a dwarf mutant, named yadokari 1-D (ydk1-D), that had a T-DNA insertion proximal to a GH3 gene. ydk1-D is dominant and has a short hypocotyl not only in light but also in darkness. Moreover, ydk1-D has a short primary root, a reduced lateral root number, and reduced apical dominance. A GH3 gene, named YDK1, was upregulated in ydk1-D, and YDK1 transgenic plants showed the ydk1-D phenotype. YDK1 gene expression was induced by exogenously applied auxin and regulated by auxin-response factor (ARF)7. In addition, YDK1 gene expression was downregulated by blue and far-red (FR) lights. Strong promoter activity of YDK1 was observed in roots and flowers. These results suggest that YDK1 may function as a negative component in auxin signaling by regulating auxin activity. PMID- 14756758 TI - Chromosome termini of the monocot plant Othocallis siberica are maintained by telomerase, which specifically synthesises vertebrate-type telomere sequences. AB - Lack of Arabidopsis-type T3AG3 telomere sequences has recently been reported for the majority of investigated taxa of the monocot order Asparagales. In order to investigate this phenomenon in more detail, we conducted extensive cytogenetic and molecular analyses of the telomeres in Othocallis siberica, a member of this order. Terminal restriction fragment analysis together with Bal31 exonuclease assay showed that chromosome termini in O. siberica are formed by long stretches (more than 10 kbp) of vertebrate-type T2AG3 repeats. In addition, telomerase activity specifically synthesising (T2AG3)n sequence was detected in O. siberica protein extracts by telomerase repeat amplification protocol (TRAP). Fluorescence in situ hybridisation (FISH) revealed the presence of the vertebrate-type T2AG3 telomere sequences at all chromosome termini and at a few additional regions of O. siberica chromosomes, whereas Arabidopsis-type T3AG3 DNA and peptide nucleic acid (PNA) probes did not hybridise to chromosomes of Othocallis, except for polymorphic blocks in chromosomes 2 (interstitial) and 4 (terminal). These interstitial/terminal regions are apparently composed of large blocks of (T2AG3)n and (T3AG3)n DNA and represent a unique example of interspersion of two types of telomeric repeats within one genome. This may be a reflection of the recent evolutionary switch from Arabidopsis- to vertebrate-type telomeric repeats in this plant group. PMID- 14756759 TI - Ethylene-mediated enhancement of apical hook formation in etiolated Arabidopsis thaliana seedlings is gibberellin dependent. AB - Dark-grown Arabidopsis seedlings develop an apical hook by differential elongation and division of hypocotyl cells. This allows the curved hypocotyl to gently drag the apex, which is protected by the cotyledons, upwards through the soil. Several plant hormones are known to be involved in hook development, including ethylene, which causes exaggeration of the hook. We show that gibberellins (GAs) are also involved in this process. Inhibition of GA biosynthesis with paclobutrazol (PAC) prevented hook formation in wild-type (WT) seedlings and in constitutive ethylene response (ctr)1-1, a mutant that exhibits a constitutive ethylene response. In addition, a GA-deficient mutant (ga1-3) did not form an apical hook in the presence of the ethylene precursor 1 aminocyclopropane-1-carboxylate (ACC). Analysis of transgenic Arabidopsis seedlings expressing a green fluorescent protein (GFP)-repressor of ga1-3 (RGA) fusion protein suggested that ACC inhibits cell elongation in the apical hook by inhibition of GA signaling. A decreased feedback of GA possibly causes an induction of GA biosynthesis based upon the expression of genes encoding copalyl diphosphate synthase (CPS; GA1) and GA 2-oxidase (AtGA2ox1). Furthermore, expression of GASA1, a GA-response gene, suggests that differential cell elongation in the apical hook might be a result of differential GA-sensitivity. PMID- 14756760 TI - Xa26, a gene conferring resistance to Xanthomonas oryzae pv. oryzae in rice, encodes an LRR receptor kinase-like protein. AB - Rice bacterial blight, caused by Xanthomonas oryzae pv. oryzae (Xoo), is one of the most serious rice diseases worldwide. A rice gene, Xa26, conferring resistance against Xoo at both seedling and adult stages was isolated by map based cloning strategies from the rice cultivar Minghui 63. Xa26 belongs to a multigene family consisting of four members. It encodes a leucine-rich repeat (LRR) receptor kinase-like protein and is constitutively expressed. Sequence analysis revealed that IRBB3 and Zhachanglong lines that are resistant to a broad range of Xoo strains, also carry Xa26. However, significant difference in lesion length was observed among these lines after inoculation with a set of Xoo strains. Moreover, transgenic plants carrying Xa26 showed enhanced resistance compared with the donor line of the gene in both seedling and adult stages. These results suggest that the resistance conferred by Xa26 is influenced by the genetic background. PMID- 14756761 TI - Genome analysis at different ploidy levels allows cloning of the powdery mildew resistance gene Pm3b from hexaploid wheat. AB - In wheat, race-specific resistance to the fungal pathogen powdery mildew (Blumeria graminis f. sp. tritici) is controlled by the Pm genes. There are 10 alleles conferring resistance at the Pm3 locus (Pm3a to Pm3j) on chromosome 1AS of hexaploid bread wheat (Triticum aestivum L.). The genome of hexaploid wheat has a size of 1.6 x 1010 bp and contains more than 80% of repetitive sequences, making positional cloning difficult. Here, we demonstrate that the combined analysis of genomes from wheat species with different ploidy levels can be exploited for positional cloning in bread wheat. We have mapped the Pm3b gene in hexaploid wheat to a genetic interval of 0.97 centimorgan (cM). The diploid T. monococcum and the tetraploid T. turgidum ssp. durum provided models for the A genome of hexaploid wheat and allowed to establish a physical contig spanning the Pm3 locus. Although the haplotypes at the Pm3 locus differed markedly between the three species, a large resistance gene-like family specific to wheat group 1 chromosomes was consistently found at the Pm3 locus. A candidate gene for Pm3b was identified using partial sequence conservation between resistant line Chul and T. monococcum cv. DV92. A susceptible Pm3b mutant, carrying a single-base pair deletion in the coding region of the candidate gene was isolated. When tested in a single cell transformation assay, the Pm3b candidate gene conferred race-specific resistance to powdery mildew. These results demonstrate that the candidate gene, a member of the coiled-coil nucleotide binding site leucine-rich repeat (NBS-LRR) type of disease resistance genes, is the Pm3b gene. PMID- 14756762 TI - Transcript profiles and deduced changes of metabolic pathways in maternal and filial tissues of developing barley grains. AB - Different aspects of barley grain development have been studied in detail, but a more global analysis of gene expression patterns is still missing. We have employed macro arrays, containing 1184 unique sequences from 1421 barley cDNA fragments, to study gene expression profiles in maternal and filial tissues of developing barley caryopses from fertilization to early storage phase. Principle component analysis (PCA) defined distinct expression networks in the pre-storage (0, 2, and 4 days after flowering (DAF)) and early storage phase (10 and 12 DAF). During an intermediate phase (6 and 8 DAF), PCA visualizes a dramatic re programming of the transcriptional machinery. In maternal tissues, a large set of protein-mobilizing enzyme mRNAs, together with upregulated lipid-mobilizing enzyme and downregulated reactive oxygen species (ROS)-scavenging enzyme genes, suggests mobilization of stored compounds and programmed cell death (PCD). In the filial tissue fraction, a set of genes highly expressed during the pre-storage phase is involved in growth processes, including cell wall biosynthesis. The data suggest that the necessary UDP-glucose is provided both by sucrose synthase (isoform 3) and an invertase-driven pathway. Further, major developmental changes in pathways producing energy are predicted. A bell-shaped expression profile with a peak during the intermediate phase is characteristic for genes associated with photosynthesis and ATP production. The photosynthesis-determined increase of ATP concentration could be a prerequisite for the initiation of grain filling, dominated by starch and storage protein synthesis. Storage product accumulation is accompanied by high transcriptional activity of genes involved in glycolysis and fermentation, as well as in the citric acid cycle. PMID- 14756763 TI - Conservation of class C function of floral organ development during 300 million years of evolution from gymnosperms to angiosperms. AB - Flower development in angiosperms is regulated by the family of MADS-box transcription factors. MADS-box genes have also been reported from gymnosperms, another major group of seed plants. AGAMOUS (AG) is the class C MADS-box floral organ identity gene controlling the stamen and carpel development in Arabidopsis. We report the characterization of an ortholog of the AG gene, named Cycas AGAMOUS (CyAG), from the primitive gymnosperm Cycas edentata. The expression pattern of CyAG in Cycas parallels that of AG in Arabidopsis. Additionally, the gene structure, including the number and location of the introns, is conserved in CyAG and other AG orthologs known. Most importantly, functional analysis shows that CyAG driven by the AG promoter can rescue the loss-of-function ag mutant of Arabidopsis. However, the ectopic expression of CyAG in ag mutant Arabidopsis cannot produce the carpeloid and stamenoid organs in the first and second whorls, although the stamen and carpel are rescued in the third and fourth whorls of the transformants. These observations show that the molecular mechanism of class C function controlling reproductive organ identity (stamen and carpel of angiosperms or microsporophyll and megasporophyll of gymnosperms) arose before the divergence of angiosperms and gymnosperms, and has been conserved during 300 million years of evolution thereafter. PMID- 14756764 TI - Cryopreservation of transformed and wild-type Arabidopsis and tobacco cell suspension cultures. AB - We have recently described Arabidopsis cell suspension cultures that can be effectively synchronised. Here, we describe procedures that allow clonal transformed cell suspension lines to be produced using Agrobacterium-mediated transformation, and an optimised and straightforward procedure for the cryopreservation and recovery of both parental and transformed lines. Frozen cultures show 90% viability and rapid re-growth after recovery. We show that the cryopreservation procedure is equally applicable to the frequently used tobacco bright yellow (BY)2 cell suspension culture, and that cell cycle synchronisation capacity of parental lines is maintained after both transformation and recovery from cryopreservation. The techniques require no specialised equipment, and are suitable for routine laboratory use, greatly facilitating the handling and maintenance of cell cultures and providing security against both contamination and cumulative somaclonal variation. Finally, the ability to store easily large numbers of transformed lines opens the possibility of using Arabidopsis cell suspension cultures for high-throughput analysis. PMID- 14756765 TI - Membrane topology and sequence requirements for oil body targeting of oleosin. AB - Oleosin protein is targeted to oil bodies via the endoplasmic reticulum (ER) and consists of a lipid-submerged hydrophobic (H) domain that is flanked by cytosolic hydrophilic domains. We investigated the relationship between oleosin ER topology and its subsequent ability to target to oil bodies. Oleosin variants were created to yield differing ER membrane topologies and tagged with a reporter enzyme. Localisation was assessed by fractionation after transient expression in embryonic cells. Membrane-straddled topologies with N-terminal sequence in the ER lumen and C-terminal sequence in the cytosol were unable to target to oil bodies efficiently. Similarly, a translocated topology with only ER membrane and lumenal sequence was unable to target to oil bodies efficiently. Both topology variants accumulated proportionately higher in ER microsomal fractions, demonstrating a block in transferring from ER to oil bodies. The residual oil body accumulation for the inverted topology was shown to be because of partial adoption of native ER membrane topology, using a reporter variant, which becomes inactivated by ER mediated glycosylation. In addition, the importance of H domain sequence for oil body targeting was assessed using variants that maintain native ER topology. The central proline knot motif (PKM) has previously been shown to be critical for oil body targeting, but here the arms of the H domain flanking this motif were shown to be interchangeable with only a moderate reduction in oil body targeting. We conclude that oil body targeting of oleosin depends on a specific ER membrane topology but does not require a specific sequence in the H domain flanking arms. PMID- 14756766 TI - The Pseudomonas syringae type III effector AvrRpt2 functions downstream or independently of SA to promote virulence on Arabidopsis thaliana. AB - AvrRpt2, a Pseudomonas syringae type III effector protein, functions from inside plant cells to promote the virulence of P. syringae pv. tomato strain DC3000 (PstDC3000) on Arabidopsis thaliana plants lacking a functional copy of the corresponding RPS2 resistance gene. In this study, we extended our understanding of AvrRpt2 virulence activity by exploring the hypothesis that AvrRpt2 promotes PstDC3000 virulence by suppressing plant defenses. When delivered by PstDC3000, AvrRpt2 suppresses pathogen-related (PR) gene expression during infection, suggesting that AvrRpt2 suppresses defenses mediated by salicylic acid (SA). However, AvrRpt2 promotes PstDC3000 growth on transgenic plants expressing the SA degrading enzyme NahG, indicating that AvrRpt2 does not promote bacterial virulence by modulating SA levels during infection. AvrRpt2 general virulence activity does not depend on the RPM1 resistance gene, as mutations in RPM1 had no effect on AvrRpt2-induced phenotypes. Transgenic plants expressing AvrRpt2 displayed enhanced susceptibility to PstDC3000 strains defective in type III secretion, indicating that enhanced susceptibility of these plants is not because of suppression of defense responses elicited by other type III effectors. Additionally, avrRpt2 transgenic plants did not exhibit increased susceptibility to Peronospora parasitica and Erysiphe cichoracearum, suggesting that AvrRpt2 virulence activity is specific to P. syringae. PMID- 14756767 TI - Identification of Pseudomonas syringae type III effectors that can suppress programmed cell death in plants and yeast. AB - The Pseudomonas syringae pv. tomato DC3000 type III secretion system (TTSS) is required for bacterial pathogenicity on plants and elicitation of the hypersensitive response (HR), a programmed cell death (PCD) that occurs on resistant plants. Cosmid pHIR11 enables non-pathogens to elicit an HR dependent upon the TTSS and the effector HopPsyA. We used pHIR11 to determine that effectors HopPtoE, avirulence AvrPphEPto, AvrPpiB1Pto, AvrPtoB, and HopPtoF could suppress a HopPsyA-dependent HR on tobacco and Arabidopsis. Mixed inoculum and Agrobacterium-mediated transient expression experiments confirmed that suppressor action occurred within plant cells. These suppressors, with the exception of AvrPpiB1Pto, inhibited the expression of the tobacco pathogenesis-related (PR) gene PR1a. DC3000 suppressor mutants elicited an enhanced HR consistent with these mutants lacking an HR suppressor. Additionally, HopPtoG was identified as a suppressor on the basis of an enhanced HR produced by a hopPtoG mutant. Remarkably, these proteins functioned to inhibit the ability of the pro-apoptotic protein, Bax to induce PCD in plants and yeast, indicating that these effectors function as anti-PCD proteins in a trans-kingdom manner. The high proportion of effectors that suppress PCD suggests that suppressing plant immunity is one of the primary roles for DC3000 effectors and a central requirement for P. syringae pathogenesis. PMID- 14756768 TI - ABA depolarizes guard cells in intact plants, through a transient activation of R and S-type anion channels. AB - During drought, the plant hormone abscisic acid (ABA) induces rapid stomatal closure and in turn reduces transpiration. Stomatal closure is accompanied by large ion fluxes across the plasma membrane, carried by K+ and anion channels. We recorded changes in the activity of these channels induced by ABA, for guard cells of intact Vicia faba plants. Guard cells in their natural environment were impaled with double-barrelled electrodes, and ABA was applied via the leaf surface. In 45 out of 85 cells tested, ABA triggered a transient depolarization of the plasma membrane. In these cells, the membrane potential partially recovered in the presence of ABA; however, a full recovery of the membrane potentials was only observed after removal of ABA. Repetitive ABA responses could be evoked in single cells, but the magnitude of the response varied from one hormone application to the other. The transient depolarization correlated with the activation of anion channels, which peaked 5 min after introduction of the stimulus. In guard cells with a moderate increase in plasma membrane conductance (DeltaG < 5 nS), ABA predominantly activated voltage-independent (slow (S)-type) anion channels. During strong responses (DeltaG > 5 nS), however, ABA activated voltage-dependent (rapid (R)-type) in addition to S-type anion channels. We conclude that the combined activation of these two channel types leads to the transient depolarization of guard cells. The nature of this ABA response correlates with the transient extrusion of Cl- from guard cells and a rapid but confined reduction in stomatal aperture. PMID- 14756769 TI - Activation of a COI1-dependent pathway in Arabidopsis by Pseudomonas syringae type III effectors and coronatine. AB - Gram-negative bacteria use a variety of virulence factors including phytotoxins, exopolysaccharides, effectors secreted by the type III secretion system, and cell wall-degrading enzymes to promote parasitism in plants. However, little is known about how these virulence factors alter plant cellular responses to promote disease. In this study, we show that virulent Pseudomonas syringae strains activate the transcription of an Arabidopsis ethylene response factor (ERF) gene, RAP2.6, in a coronatine insensitive 1 (COI1)-dependent manner. A highly sensitive RAP2.6 promoter-firefly luciferase (RAP2.6-LUC) reporter line was developed to monitor activities of various bacterial virulence genes. Analyses of P. syringae pv. tomato DC3000 mutants indicated that both type III secretion system and the phytotoxin coronatine are required for RAP2.6 induction. We show that at least five individual type III effectors, avirulence B (AvrB), AvrRpt2, AvrPphB, HopPtoK, and AvrPphEPto, contributed to RAP2.6 induction. Gene-for-gene recognition was not involved in RAP2.6 induction because plants lacking RPM1 and RPS2 responded normally to AvrB and AvrRpt2 in RAP2.6 expression. Interestingly, the role of coronatine in RAP2.6 induction can be partially substituted by the addition of avrB in DC3000, suggesting that AvrB may mimic coronatine. These results suggest that P. syringae type III effectors and coronatine act by augmenting a COI1-dependent pathway to promote parasitism. PMID- 14756770 TI - Forest tent caterpillars (Malacosoma disstria) induce local and systemic diurnal emissions of terpenoid volatiles in hybrid poplar (Populus trichocarpa x deltoides): cDNA cloning, functional characterization, and patterns of gene expression of (-)-germacrene D synthase, PtdTPS1. AB - Feeding forest tent caterpillars (FTCs) induced local and systemic diurnal emissions of (-)-germacrene D, along with (E)-beta-ocimene, linalool, (E)-4,8 dimethyl-1,3,7-nonatriene (DMNT), benzene cyanide, and (E,E)-alpha-farnesene, from leaves of hybrid poplar. FTC feeding induced substantially higher levels of volatiles in local and systemic leaves than did mechanical wounding. A full length poplar sesquiterpene synthase cDNA (PtdTPS1) was isolated and functionally identified as (-)-germacrene D synthase. Expression of PtdTPS1, expression of genes of early, intermediate and late steps in terpenoid biosynthesis, and expression of a lipoxygenase gene (PtdLOX1) were analyzed in local FTC-infested and systemic leaves. Transcript levels of PtdTPS1 and PtdLOX1 were strongly increased in response to herbivory. PtdTPS1 was also induced by mechanical wounding or by methyl jasmonate (MeJA) treatment. FTC feeding did not affect transcript levels of 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR), 1-deoxy-d xylulose 5-phosphate reductoisomerase (DXR), and isoprene synthase (IPS). Two other TPS genes, PtdTPS2 and PtTPS3, and farnesyl diphosphate synthase were only very transiently induced. These results illustrate differential expression of terpenoid pathway genes in response to insect feeding and a key function of (-) germacrene D synthase PtdTPS1 for herbivore-induced local and systemic volatile emissions in hybrid poplar. FTC-induced transcripts of PtdTPS1 followed diurnal rhythm. Spatial patterns of FTC-induced PtdTPS1 transcript accumulation revealed acropetal but not basipetal direction of the systemic response. Implications for tritrophic poplar-FTC-predator/parasitoid interactions are discussed. PMID- 14756771 TI - Glycerol-insensitive Arabidopsis mutants: gli1 seedlings lack glycerol kinase, accumulate glycerol and are more resistant to abiotic stress. AB - The aim of this study was to investigate the process of glycerol catabolism in germinating Arabidopsis seed. A genetic screen was performed to isolate glycerol insensitive (gli) mutant seedlings. Three separate mutant loci were identified (gli1, gli2 and gli3). Of these, only gli1 is unable to utilise glycerol. Following germination, gli1 seedlings transiently accumulate glycerol derived from the breakdown of storage oil and are more resistant to hyperosmotic stress, salt stress, oxidative stress, freezing and desiccation. Enzyme assays revealed that gli1 lacks glycerol kinase activity. GLI1 mapped to chromosome 1 near the putative glycerol kinase gene NHO1. Mutations in this gene were identified in three independent gli1 alleles. A cDNA encoding GLI1 was cloned and its function was proven by complementation of an Escherichia coli glycerol kinase (glpK) deletion strain. Quantitative RT-PCR analysis showed that GLI1 is expressed in all tissues, but is transiently upregulated during early post-germinative growth and leaf senescence. These data show that glycerol kinase is required for glycerol catabolism in Arabidopsis and that the accumulation of glycerol can enhance resistance to a variety of abiotic stresses associated with dehydration. PMID- 14756772 TI - GID2, an F-box subunit of the SCF E3 complex, specifically interacts with phosphorylated SLR1 protein and regulates the gibberellin-dependent degradation of SLR1 in rice. AB - The phytohormone gibberellin (GA) controls growth and development in plants. Previously, we identified a rice F-box protein, gibberellin-insensitive dwarf2 (GID2), which is essential for GA-mediated DELLA protein degradation. In this study, we analyzed the biological and molecular biological properties of GID2. Expression of GID2 preferentially occurred in rice organs actively synthesizing GA. Domain analysis of GID2 revealed that the C-terminal regions were essential for the GID2 function, but not the N-terminal region. Yeast two-hybrid assay and immunoprecipitation experiments demonstrated that GID2 is a component of the SCF complex through an interaction with a rice ASK1 homolog, OsSkp15. Furthermore, an in vitro pull-down assay revealed that GID2 specifically interacted with the phosphorylated Slender Rice 1 (SLR1). Taken these results together, we conclude that the phosphorylated SLR1 is caught by the SCFGID2 complex through an interacting affinity between GID2 and phosphorylated SLR1, triggering the ubiquitin-mediated degradation of SLR1. PMID- 14756773 TI - Evolution of fungal sex chromosomes. AB - Sexual reproduction enables organisms to shuffle two parental genomes to produce recombinant progeny, and to purge the genome of deleterious mutations. Sex is conserved in virtually all organisms, from bacteria and fungi to plants and animals, and yet the mechanisms by which sexual identity are established share both conserved general features and are remarkably diverse. In animals, sexual identity is established by dimorphic sex chromosomes, whereas in fungi a specialized region of the genome, known as the mating-type locus, governs the establishment of cell type identity and differs in DNA sequence between cells of different mating-types. Recent studies on the mating-type loci of fungi and algae reveal features shared with the mammalian X and Y chromosomes, suggesting that these represent early steps in the evolution of sex chromosomes. PMID- 14756774 TI - Redundancy in the function of mitochondrial phosphate transport in Saccharomyces cerevisiae and Arabidopsis thaliana. AB - Most cellular ATP is produced within the mitochondria from ADP and Pi which are delivered across the inner-membrane by specific nuclearly encoded polytopic carriers. In Saccharomyces cerevisiae, some of these carriers and in particular the ADP/ATP carrier, are represented by several related isoforms that are distinct in their pattern of expression. Until now, only one mitochondrial Pi carrier (mPic) form, encoded by the MIR1 gene in S. cerevisiae, has been described. Here we show that the gene product encoded by the YER053C ORF also participates in the delivery of phosphate to the mitochondria. We have called this gene PIC2 for Pi carrier isoform 2. Overexpression of PIC2 compensates for the mitochondrial defect of the double mutant Deltamir1 Deltapic2 and restores phosphate transport activity in mitochondria swelling experiments. The existence of two isoforms of mPic does not seem to be restricted to S. cerevisiae as two Arabidopsis thaliana cDNAs encoding two different mPic-like proteins are also able to complement the double mutant Deltamir1 Deltapic2. Finally, we demonstrate that Pic2p is a mitochondrial protein and that its steady state level increases at high temperature. We propose that Pic2p is a minor form of mPic which plays a role under specific stress conditions. PMID- 14756775 TI - From the inside out--processing of the Chlamydial autotransporter PmpD and its role in bacterial adhesion and activation of human host cells. AB - Polymorphic membrane protein (Pmp)21 otherwise known as PmpD is the longest of 21 Pmps expressed by Chlamydophila pneumoniae. Recent bioinformatical analyses annotated PmpD as belonging to a family of exported Gram-negative bacterial proteins designated autotransporters. This prediction, however, was never experimentally supported, nor was the function of PmpD known. Here, using 1D and 2D PAGE we demonstrate that PmpD is processed into two parts, N-terminal (N pmpD), middle (M-pmpD) and presumably third, C-terminal part (C-pmpD). Based on localization of the external part on the outer membrane as shown by immunofluorescence, immuno-electron microscopy and immunoblotting combined with trypsinization, we demonstrate that N-pmpD translocates to the surface of bacteria where it non-covalently binds other components of the outer membrane. We propose that N-pmpD functions as an adhesin, as antibodies raised against N-pmpD blocked chlamydial infectivity in the epithelial cells. In addition, recombinant N-pmpD activated human monocytes in vitro by upregulating their metabolic activity and by stimulating IL-8 release in a dose-dependent manner. These results demonstrate that N-PmpD is an autotransporter component of chlamydial outer membrane, important for bacterial invasion and host inflammation. PMID- 14756776 TI - Flavonoids induce temporal shifts in gene-expression of nod-box controlled loci in Rhizobium sp. NGR234. AB - Rhizobia, soil bacteria of the Rhizobiales, enter the roots of homologous legumes, where they induce the formation of nitrogen-fixing nodules. Signals emanating from both symbiotic partners control nodule development. Efficient nodulation requires precise, temporal regulation of symbiotic genes. Roots continuously release flavonoids that interact with transcriptional activators of the LysR family. NodD proteins, which are members of this family, act both as sensors of the environment and modulate the expression of genes preceded by conserved promoter sequences called nod-boxes. The symbiotic plasmid of the broad host-range Rhizobium sp. NGR234 caries 19 nod-boxes (NB1 to NB19), all of which were cloned upstream of a lacZ-reporter gene. A flavonoid, daidzein was able to induce 18 of the 19 nod-boxes in a NodD1-dependent manner. Interestingly, induction of four nod-boxes (NB6, NB15, NB16 and NB17) is highly dependent on NodD2 and was delayed in comparison with the others. In turn, NodD2 is involved in the repression of the NB8 nodABCIJnolOnoeI operon. Activation of transcription of nodD2 is also dependent on flavonoids despite the absence of a nod-box like sequence in the upstream promoter region. Mutational analysis showed that syrM 2 (another member of the LysR family), which is controlled by NB19, is also necessary for expression of nodD 2. Thus, NodD1, NodD2 and SyrM2 co-modulate a flavonoid-inducible regulatory cascade that coordinates the expression of symbiotic genes with nodule development. PMID- 14756777 TI - Hyperinitiation of DNA replication in Escherichia coli leads to replication fork collapse and inviability. AB - Elevated dnaA expression from a multicopy plasmid induces more frequent initiation from the Escherichia coli replication origin, oriC, but viability is maintained. In comparison, chromosomally encoded dnaAcos also stimulates initiation, but this is lethal. By quantitative methods, we show that the level of initiation induced by elevated dnaA expression leads to collapsed replication forks that are mostly within 10 map units of oriC. Because forks collapse randomly, nucleoprotein complexes at specific sites such as datA are not the cause. When replication restart is blocked by a mutation in recB or priA, the increased initiations via elevated dnaA expression causes inviability. The amount of collapsed forks is substantially higher under elevated expression of dnaAcos compared to that of dnaA. We propose that the lethal phenotype of chromosomally encoded dnaAcos is a result of hyperinitiation that overwhelms the repair capacity of the cell. PMID- 14756778 TI - Individual RD1-region genes are required for export of ESAT-6/CFP-10 and for virulence of Mycobacterium tuberculosis. AB - The RD1 genomic region is present in virulent strains of Mycobacterium tuberculosis (MTB), missing from the vaccine strain M. bovis BCG, and its importance to virulence has been established experimentally. Based on in silico analysis, it has been suggested that RD1 may encode a novel secretion system, but the mechanism by which this region affects virulence is unknown. Here we examined mutants disrupted in five individual RD1 genes. Both in vitro and in vivo, each mutant displayed an attenuated phenotype very similar to a mutant missing the entire RD1 region. Genetic complementation of individual genes restored virulence. Attenuated mutants could multiply within THP-1 cells, but they were unable to spread to uninfected macrophages. We also examined export of two immunodominant RD1 proteins, CFP-10 and ESAT-6. Export of these proteins was greatly reduced or abolished in each attenuated mutant. Again, genetic complementation restored a wild-type phenotype. Our results indicate that RD1 genes work together to form a single virulence determinant, and argue that RD1 encodes a novel specialized secretion system that is required for pathogenesis of MTB. PMID- 14756779 TI - An ER packaging chaperone determines the amino acid uptake capacity and virulence of Candida albicans. AB - The Candida albicans CSH3 gene encodes a functional and structural homologue of Shr3p, a yeast protein that is specifically required for proper uptake and sensing of extracellular amino acids in Saccharomyces cerevisiae. A Candida csh3delta/csh3delta null mutant has a reduced capacity to take up amino acids, and is unable to switch morphologies on solid and in liquid media in response to inducing amino acids. CSH3/csh3delta heterozygous strains display normal amino acid induced morphological switching. However, although heterozygous cells apparently sense and properly react to amino acid induced signals they cannot take up amino acids at wild-type rates. Strikingly, both CSH3/csh3delta heterozygous and csh3delta/csh3delta homozygous strains are unable to efficiently mount virulent infections in a mouse model. The haploinsufficiency phenotypes indicate that both CSH3 alleles contribute to maintain high-capacity amino acid uptake in wild-type strains. These results strongly suggest that C. albicans cells use amino acids, presumably as nitrogen sources, during growth in mammalian hosts. PMID- 14756780 TI - Host processing of branched DNA intermediates is involved in targeted transposition of IS911. AB - A simplified system using bacterial insertion sequence IS911 has been developed to investigate targeted insertion next to DNA sequences resembling IS ends. We show here that these IR-targeted events occur by an unusual mechanism. In the circular IS911 transposition intermediate the two IRs are abutted to form an IR/IR junction. IR-targeted insertion involves transfer of a single end of the junction to the target IR to generate a branched DNA structure. The single-end transfer (SET) intermediate, but not the final insertion product, can be detected in an in vitro reaction. SET intermediates must be processed by the bacterial host to obtain the final insertion products. Sequence analysis of these IR targeted insertion products and of those obtained in vivo revealed high levels of DNA sequence conversion in which mutations from one IR were transferred to another. These sequence changes cannot be explained by the classic transposition pathway. A model is presented in which the four-way Holliday-like junction created by SET is processed by host-mediated branch migration, resolution, repair and replication. This pathway resembles those described for processing other branched DNA structures such as stalled replication forks. PMID- 14756781 TI - Nucleoid restructuring in stationary-state bacteria. AB - The textbook view of the bacterial cytoplasm as an unstructured environment has been overturned recently by studies that highlighted the extent to which non random organization and coherent motion of intracellular components are central for bacterial life-sustaining activities. Because such a dynamic order critically depends on continuous consumption of energy, it cannot be perpetuated in starved, and hence energy-depleted, stationary-state bacteria. Here, we show that, at the onset of the stationary state, bacterial chromatin undergoes a massive reorganization into ordered toroidal structures through a process that is dictated by the intrinsic properties of DNA and by the ubiquitous starvation induced DNA-binding protein Dps. As starvation proceeds, the toroidal morphology acts as a structural template that promotes the formation of DNA-Dps crystalline assemblies through epitaxial growth. Within the resulting condensed assemblies, DNA is effectively protected by means of structural sequestration. We thus conclude that the transition from bacterial active growth to stationary phase entails a co-ordinated process, in which the energy-dependent dynamic order of the chromatin is sequentially substituted with an equilibrium crystalline order. PMID- 14756782 TI - Bacillus anthracis requires siderophore biosynthesis for growth in macrophages and mouse virulence. AB - Systemic anthrax infections can be characterized as proceeding in stages, beginning with an early intracellular establishment stage within phagocytes that is followed by extracelluar stages involving massive bacteraemia, sepsis and death. Because most bacteria require iron, and the host limits iron availability through homeostatic mechanisms, we hypothesized that B. anthracis requires a high affinity mechanism of iron acquisition during its growth stages. Two putative types of siderophore synthesis operons, named Bacillus anthracis catechol, bac (anthrabactin), and anthrax siderophore biosynthesis, asb (anthrachelin), were identified. Directed gene deletions in both anthrabactin and anthrachelin pathways were generated in a B. anthracis (Sterne) 34F2 background resulting in mutations in asbA and bacCEBF. A decrease in siderophore production was observed during iron-depleted growth in both the DeltaasbA and DeltabacCEBF strains, but only the DeltaasbA strain was attenuated for growth under these conditions. In addition, the DeltaasbA strain was severely attenuated both for growth in macrophages (MPhi) and for virulence in mice. In contrast, the DeltabacCEBF strain did not differ phenotypically from the parental strain. These findings support a requirement for anthrachelin but not anthrabactin in iron assimilation during the intracellular stage of anthrax. PMID- 14756783 TI - Thermoregulation of the Escherichia coli O157:H7 pO157 ecf operon and lipid A myristoyl transferase activity involves intrinsically curved DNA. AB - Escherichia coli O157:H7 survives in diverse environments from the ruminant gastrointestinal tract to cool nutrient-dilute water. We hypothesized that the gene regulation required for this flexibility includes intrinsically curved DNA that responds to environmental changes. Three intrinsically curved DNAs were cloned from the E. coli O157:H7 virulence plasmid (pO157), sequenced and designated Bent 1 through Bent 3 (BNT1, BNT2 and BNT3). Compared to BNT1 and BNT3, BNT2 had characteristics typical of intrinsically curved DNA including electrophoretic gel retardation at 4 degrees C, six partially phased adenine:thymine tracts and transcriptional activation. BNT2::lacZ operon fusions showed that BNT2 activated transcription at 24 degrees C compared to 37 degrees C and was partially repressed by a bacterial nucleoid-associated protein H-NS. BNT2 regulated the E. coli attaching and effacing gene-positive conserved fragments 1 4 (ecf1-4) that are conserved in Shiga toxin-producing E. coli associated with human disease. Experimental analyses showed that ecf1-4 formed an operon. ecf1, 2 and 3 encoded putative proteins associated with bacterial surface polysaccharide biosynthesis and invasion and ecf4 complemented a chromosomal deletion of lpxM encoding lipid A myristoyl transferase. Mass spectrometric analysis of lipid A from ecf and lpxM single and double mutants showed that myristoylation was altered at lower temperature. PMID- 14756784 TI - Evidence that putrescine acts as an extracellular signal required for swarming in Proteus mirabilis. AB - In a search for Proteus mirabilis genes that were regulated by cell-to-cell signalling, a lacZ fusion (cmr437::mini-Tn5lacZ) was identified that was repressed 10-fold by a self-produced extracellular signal from wild-type cells. However, the cmr437::mini-Tn5lacZ insertion itself led to a marked reduction in this extracellular repressing signal. The cmr437::mini-Tn5lacZ insertion was mapped to a speA homologue in P. mirabilis. Sequence analysis indicated that a speB homologue was encoded downstream of speA. Products of the SpeA and SpeB enzymes (agmatine and putrescine) were tested for repression of cmr437::lacZ. Agmatine did not have repressing activity. However, putrescine was an effective repressing molecule at concentrations down to 30 microM. A second prominent phenotype of the cmr437 (speA)::mini-Tn5lacZ insertion was a severe defect in swarming motility. This swarming defect was also observed in a strain containing a disruption of the downstream speB gene. Differentiation of the speB mutant to swarmer cells was delayed by two hours relative to wild-type cells. Furthermore, the speB mutant was unable to migrate effectively across agar surfaces and formed very closely spaced swarming rings. Exogenous putrescine restored both the normal timing of swarmer cell differentiation and the ability to migrate to speB mutants. PMID- 14756785 TI - Mutations of the CK2 phosphorylation site of Sic1 affect cell size and S-Cdk kinase activity in Saccharomyces cerevisiae. AB - By sequence analysis we found an amino acid stretch centred on Serine201 matching a stringent CK2 consensus site within the C-terminal, inhibitory domain of Sic1. Here we show by direct mass spectrometry analysis that Sic1, but not a mutant protein whose CK2 phospho-acceptor site has been mutated to alanine, Sic1S201A, is actually phosphorylated in vitro by CK2 on Serine 201. Mutation of Serine 201 alters the coordination between growth and cell cycle progression. A significant increase of average protein content and of the average protein content at the onset of DNA synthesis is observed for exponentially growing cells harbouring the Sic1S201A protein. A strong reduction of the same parameters is observed in cells harbouring Sic1S201E. The deregulated coordination between cell size and cell cycle is also apparent at the level of S-Cdk activity. PMID- 14756786 TI - Kinetics of plasmid segregation in Escherichia coli. AB - Low copy-number bacterial replicons occupy specific locations in their host cells. Production of a GFP-Lac repressor hybrid protein in cells carrying F or P1 plasmids tagged with a lac operator array reveals that in smaller (younger) cells these plasmids are seen mainly as a single fluorescent focus at mid-cell, whereas larger cells tend to have two foci, one at each quarter-cell position. Duplication of the central focus is presumed to represent active partition of plasmid copies. We report here our investigation by time-lapse microscopy of the subsequent movement of these copies to the quarter positions. Following duplication of the central focus, the new foci migrated rapidly and directly to their quarter-cell destinations, where they remained until the next cell cycle. The speed of movement was about five times faster than poleward migration of oriC and 50 times faster than cell elongation. Aberrant positioning of mini-F lacking its sopC centromere demonstrated the requirement for the partition system in this localization process. From the measured number of F plasmid copies per cell it appears that each migrating focus contains two or more plasmid molecules. The molecular basis of this clustering, and evidence for phasing of the partition event in the cell cycle, are discussed. PMID- 14756787 TI - A CDP-choline pathway for phosphatidylcholine biosynthesis in Treponema denticola. AB - The genomes of Treponema denticola and Treponema pallidum contain a gene, licCA, which is predicted to encode a fusion protein containing choline kinase and CTP:phosphocholine cytidylyltransferase activities. Because both organisms have been reported to contain phosphatidylcholine, this raises the possibility that they use a CDP-choline pathway for the biosynthesis of phosphatidylcholine. This report shows that phosphatidylcholine is a major phospholipid in T. denticola, accounting for 35-40% of total phospholipid. This organism readily incorporated [14C]choline into phosphatidylcholine, indicating the presence of a choline dependent biosynthetic pathway. The licCA gene was cloned, and recombinant LicCA had choline kinase and CTP:phosphocholine cytidylyltransferase activity. The licCA gene was disrupted in T. denticola by erythromycin cassette mutagenesis, resulting in a viable mutant. This disruption completely blocked incorporation of either [14C]choline or 32Pi into phosphatidylcholine. The rate of production of another phospholipid in T. denticola, phosphatidylethanolamine, was elevated considerably in the licCA mutant, suggesting that the elevated level of this lipid compensated for the loss of phosphatidylcholine in the membranes. Thus it appears that T. denticola does contain a licCA-dependent CDP-choline pathway for phosphatidylcholine biosynthesis. PMID- 14756788 TI - Salmonella type III secretion-associated chaperones confer secretion-pathway specificity. AB - Type III protein secretion systems (TTSSs) are ancestrally related to the flagellar export system and are essential for the virulence of many bacteria pathogenic for humans, animals and plants. Most proteins destined to travel the TTSS pathway possess at least two domains that specifically target them to the secretion apparatus. One of the domains is located within the amino terminal first approximately 20 amino acids and the second domain, located within the first approximately 140 amino acids, serves as a binding site for specific chaperones. It has been previously proposed that these two secretion signals are capable of operating independently of one another to facilitate secretion into the extracellular environment. We have found that in the absence of their chaperone-binding domains, the Salmonella typhimurium TTSS-secreted proteins SptP and SopE are no longer targeted for secretion through their cognate TTSS and, instead, are secreted through the flagellar export pathway. These results indicate the existence of an 'ancestral' flagellar secretion signal within TTSS exported proteins that is revealed in the absence of the chaperone-binding domain. Furthermore, we found that secretion into culture supernatants as well as translocation into host cells by the cognate TTSS require both, the amino terminal and chaperone-binding domains. We conclude from these studies that a critical function for the TTSS-associated chaperones is to confer secretion pathway specificity to their cognate secreted proteins. PMID- 14756789 TI - Staphylococcus aureus multiresistance plasmid pSK41: analysis of the replication region, initiator protein binding and antisense RNA regulation. AB - The vast majority of large staphylococcal plasmids characterized to date appear to possess an evolutionarily common replication system, which has clearly had a major impact on the evolution of antimicrobial resistant staphylococci worldwide. Related systems have also been found in plasmids from other Gram-positive genera, including enterococci, streptococci and bacilli. The 46.4 kb plasmid pSK41 is the prototype of a family of conjugative staphylococcal multiresistance plasmids. The replication region of pSK41 encodes a protein product, Rep, which was shown to be essential for replication; mutations that truncated Rep could be complemented in trans. Rep was found to bind in vitro to four tandem repeat sequences located centrally within the rep coding region. An A + T-rich inverted repeat sequence upstream of rep was required for efficient replication, whereas no sequences downstream of rep were necessary. An antisense countertranscript, RNAI, encoded upstream of rep was identified and transcriptional start points for both RNAI and the rep-mRNA were defined. PMID- 14756790 TI - Chromosomal constraints in Gram-positive bacteria revealed by artificial inversions. AB - We used artificial chromosome inversions to investigate the chromosomal constraints that preserve genome organization in the Gram-positive bacterium Lactococcus lactis. Large inversions, 80-1260 kb in length, disturbing the symmetry of the origin and terminus of the replication axis to various extents, were constructed using the site-specific Cre-loxP recombination system. These inversions were all mechanistically feasible and fell into various classes according to stability and effect on cell fitness. The L. lactis chromosome supports only to some extent unbalance in length of its replication arms. The location of detrimental inversions allowed identification of two constrained chromosomal regions: a large domain covering one fifth of the genome that encompasses the origin of replication (Ori domain), and a smaller domain located at the opposite of the chromosome (Ter domain). PMID- 14756791 TI - The virF promoter in Shigella: more than just a curved DNA stretch. AB - In the human enteropathogen Shigella transcription of virF, the primary regulator of the invasion functions, is strictly temperature-dependent and is antagonistically mediated by H-NS and FIS, which bind to specific sites on the virF promoter. Here we report on the relevance of DNA geometry to the thermoregulation of virF and demonstrate that the virF promoter hosts a major DNA bend halfway between two H-NS sites. The bent region has been mutagenized in vitro to mimic temperature-induced changes of DNA curvature. Functional analysis of curvature mutants and of promoter constructs in which the two H-NS sites are phased-out by a half-helix turn reveals that modifying the spatial relationships between these sites severely affects the interaction of H-NS with the virF promoter, as well as its in vivo and in vitro temperature-dependent activity. The role of promoter curvature as thermosensor is also compatible with the present observation that, with increasing temperature, the virF bending centre moves downstream at a rate having its maximum around the transition temperature, abruptly unmasking a binding site for the transcriptional activator FIS. PMID- 14756793 TI - Chromosome 1 trisomy compromises the virulence of Candida albicans. AB - Although increases in chromosome copy number typically have devastating developmental consequences in mammals, fungal cells such as Saccharomyces cerevisiae seem to tolerate trisomies without obvious impairment of growth. Here, we demonstrate that two commonly used laboratory strains of the yeast Candida albicans, CAI-4 and SGY-243, can carry three copies of chromosome 1. Although the trisomic strains grow well in the laboratory, Ura+ derivatives of CAI-4, carrying three copies of chromosome 1, are avirulent in the intravenously inoculated mouse model, unlike closely related strains carrying two copies of chromosome 1. Furthermore, changes in chromosome copy number occur during growth in an animal host and during growth in the presence of growth-inhibiting drugs. These results suggest that chromosome copy number variation provides a mechanism for genetic variation in this asexual organism. PMID- 14756792 TI - Molybdate transport and its effect on nitrogen utilization in the cyanobacterium Anabaena variabilis ATCC 29413. AB - Molybdenum is an essential component of the cofactors of many metalloenzymes including nitrate reductase and Mo-nitrogenase. The cyanobacterium Anabaena variabilis ATCC 29413 uses nitrate and atmospheric N2 as sources of nitrogen for growth. Two of the three nitrogenases in this strain are Mo-dependent enzymes, as is nitrate reductase; thus, transport of molybdate is important for growth of this strain. High-affinity transport of molybdate in A. variabilis was mediated by an ABC-type transport system encoded by the products of modA and modBC. The modBC gene comprised a fused orf including components corresponding to modB and modC of Escherichia coli. The deduced ModC part of the fused gene lacked a recognizable molybdate-binding domain. Expression of modA and modBC was induced by starvation for molybdate. Mutants in modA or modBC were unable to grow using nitrate or Mo-nitrogenase. Growth using the alternative V-nitrogenase was not impaired in the mutants. A high concentration of molybdate (10 microM) supported normal growth of the modBC mutant using the Nif1 Mo-nitrogenase, indicating that there was a low-affinity molybdate transport system in this strain. The modBC mutant did not detectably transport low concentrations of 99Mo (molybdate), but did transport high concentrations. However, such transport was observed only after cells were starved for sulphate, suggesting that an inducible sulphate transport system might also serve as a low-affinity molybdate transport system in this strain. PMID- 14756794 TI - RcaE is a complementary chromatic adaptation photoreceptor required for green and red light responsiveness. AB - The recent discovery of large numbers of phytochrome photoreceptor genes in both photosynthetic and non-photosynthetic prokaryotes has led to efforts to understand their physiological roles in environmental acclimation. One receptor in this class, RcaE, is involved in controlling complementary chromatic adaptation, a process that regulates the transcription of operons encoding light harvesting proteins in cyanobacteria. Although all previously identified phytochrome responses are maximally sensitive to red and far red light, complementary chromatic adaptation is unique in that it is responsive to green and red light. Here, we present biochemical and genetic evidence demonstrating that RcaE is a photoreceptor and that it requires the cysteine at position 198 to ligate an open chain tetrapyrrole covalently in a manner analogous to chromophore attachment in plant phytochromes. Furthermore, although the wild-type rcaE gene can rescue red and green light photoresponses of an rcaE null mutant, a gene in which the codon for cysteine 198 is converted to an alanine codon rescues the red light but not the green light response. Thus, RcaE is a photoreceptor that is required for both green and red light responsiveness during complementary chromatic adaptation and is the first identified phytochrome class sensor that is involved in sensing and responding to green and red light rather than red and far red light. PMID- 14756795 TI - In vivo studies on putative Shine-Dalgarno sequences of the halophilic archaeon Halobacterium salinarum. AB - The involvement of Shine-Dalgarno sequences in the translation of mRNA in halophilic archaea was investigated for two gvp genes involved in gas vesicle formation in Halobacterium salinarum PHH1. With the exception of gvpA and gvpO, all reading frames of the p-gvpDEFGHIJKLM and p-gvpACNO mRNAs contained upstream of the AUG start codon a putative Shine-Dalgarno (SD) sequence that is complementary to the 3'-end of the small ribosomal subunit RNA. The importance of the SD sequences of gvpG and gvpH was investigated in Haloferax volcanii transformants, and an alteration of the SD sequence resulted in a reduction of the amount of the GvpG or GvpH protein. For a more quantitative analysis the region upstream of gvpH was fused to the bgaH reading frame encoding an enzyme with beta-galactosidase activity as reporter. Scanning mutagenesis within the mRNA leader demonstrated that mutations adjacent to the putative SD sequence GGAGGUCA did not influence the efficiency of translation, whereas constructs harbouring an altered SD sequence yielded only 5-50% of the beta-galactosidase activities obtained with the wild-type SD element. A complete mutation of the SD sequence still yielded 20% of the wild-type activity. Alterations in the spacing of the SD sequence and the translation initiation codon of gvpH indicated that a distance of 4 or 10 nucleotides yielded a similar beta-galactosidase activity as found with the 7 nt spacing of the SD element in wild type, whereas a distance of 1 nt resulted in the loss of translation. A complete deletion of the 5'-UTR resulting in a leaderless mRNA yielded an enhanced beta-galactosidase activity in transformants implying that the initiation of translation involved a mechanism other than a specific mRNA-rRNA interaction. PMID- 14756796 TI - Cellulolysis is severely affected in Clostridium cellulolyticum strain cipCMut1. AB - Progress towards understanding the molecular basis of cellulolysis by Clostridium cellulolyticm was obtained through the study of the first cellulolysis defective mutant strain, namely cipCMut1. In this mutant, a 2 659 bp insertion element, disrupts the cipC gene at the sequence encoding the seventh cohesin of the scaffoldin CipC. cipC is the first gene in a large 'cel' gene cluster, encoding several enzymatic subunits of the cellulosomes, including the processive cellulase Cel48F, which is the major component. Physiological and biochemical studies showed that the mutant strain was affected in cellulosome synthesis and severely impaired in its ability to degrade crystalline cellulose. It produced small amounts of a truncated CipC protein (P120), which had functional cohesin domains and assembled complexes which did not contain any of the enzymes encoded by genes of the 'cel' cluster. The mutant cellulolytic system was mainly composed of three proteins designated P98, P105 and P125. Their N-termini did not match any of the known cellulase sequences from C. cellulolyticum. A large amount of entire CipC produced in the cipCMut1 strain by trans-complementation with plasmid pSOScipC did not restore the cellulolytic phenotype, in spite of the assembly of a larger amount of complexes. The complexes produced in the mutant and complemented strains contained at least 12 different dockerin-containing proteins encoded by genes located outside of the 'cel' cluster. The disturbances observed in the mutant and trans-complemented strains were the result of a strong polar effect resulting from the cipC gene disruption. In conclusion, this study provided genetic evidence that the cellulases encoded by the genes located in the 'cel' cluster are essential for the building of cellulosomes efficient in crystalline cellulose degradation. PMID- 14756797 TI - Use of antisense RNA to modify the composition of cellulosomes produced by Clostridium cellulolyticum. AB - The enzymatic composition of the cellulosomes produced by Clostridium cellulolyticum was modified by inhibiting the synthesis of Cel48F that is the major cellulase of the cellulosomes. The strain ATCC 35319 (pSOSasrF) was developed to over-produce a 469 nucleotide-long antisense-RNA (asRNA) directed against the ribosome-binding site region and the beginning of the coding region of the cel48F mRNAs. The cellulolytic system secreted by the asRNA-producing strain showed a markedly lower amount of Cel48F, compared to the control strain transformed with the empty plasmid (pSOSzero). This was correlated with a 30% decrease of the specific activity of the cellulolytic system on Avicel cellulose, indicating that Cel48F plays an important role in the recalcitrant cellulose degradation. However, only minor effects were observed on the growth parameters on cellulose. In both transformant strains, cellulosome production was found to be reduced and two unknown proteins (P105 and P98) appeared as major components of their cellulolytic systems. These proteins did not contain any dockerin domain and were shown to be not included into the cellulosomes; they are expected to participate to the non-cellulosomal cellulolytic system of C. cellulolyticum. PMID- 14756798 TI - PrPc on the road: trafficking of the cellular prion protein. AB - The glycosylphosphatidylinositol (GPI)-anchored cellular prion protein (PrPc) has a fundamental role in prion diseases. Intracellular trafficking of PrPc is important in the generation of protease resistant PrP species but little is known of how endocytosis affects PrPc function. Here, we discuss recent experiments that have illuminated how PrPc is internalized and what are the possible destinations taken by the protein. Contrary to what would be expected for a GPI anchored protein there is increasing evidence that clathrin-mediated endocytosis and classical endocytic organelles participate in PrPc trafficking. Moreover, the N-terminal domain of PrPc may be involved in sorting events that can direct the protein during its intracellular journey. Indeed, the concept that the GPI-anchor determines PrPc trafficking has been challenged. Cellular signaling can be triggered or be regulated by PrPc and we suggest that endocytosis of PrPc may influence signaling in several ways. Definition of the processes that participate in PrPc endocytosis and intracellular trafficking can have a major impact on our understanding of the mechanisms involved in PrPc function and conversion to protease resistant conformations. PMID- 14756799 TI - Potassium, sodium, calcium and glutamate-gated channels: pore architecture and ligand action. AB - In the last decade, the idea of common organization of certain ion channel families exhibiting diverse physiological and pharmacological properties has received strong experimental support. Transmembrane topologies and patterns of the pore-facing residues are conserved in P-loop channels that include high selective cation channels and certain ligand-gated channels. X-ray structures of bacterial K+ channels, KcsA, MthK and KvAP, help to understand structure-function relationships of other P-loop channels. Data on binding sites and mechanisms of action of ligands of K+, Na+, Ca2+ and glutamate gated ion channels are considered in view of their possible structural similarity to the bacterial K+ channels. Emphasized are structural determinants of ligand-receptor interactions within the channels and mechanisms of state-dependent action of the ligands. PMID- 14756800 TI - Parkinson's disease transgenic mitochondrial cybrids generate Lewy inclusion bodies. AB - Many models of Parkinson's disease (PD) have succeeded in replicating dopaminergic neuron loss or alpha-synuclein aggregation but not the formation of classical Lewy bodies, the pathological hallmark of PD. Our cybrid model of sporadic PD was created by introducing the mitochondrial genes from PD patients into neuroblastoma cells that lack mitochondrial DNA. Previous studies using cybrids have shown that information encoded by mitochondrial DNA in patients contributes to many pathogenic features of sporadic PD. In this paper, we report the generation of fibrillar and vesicular inclusions in a long-term cybrid cell culture model that replicates the essential antigenic and structural features of Lewy bodies in PD brain without the need for exogenous protein expression or inhibition of mitochondrial or proteasomal function. The inclusions generated by PD cybrid cells stained with eosin, thioflavin S, and antibodies to alpha synuclein, ubiquitin, parkin, synphilin-1, neurofilament, beta-tubulin, the proteasome, nitrotyrosine, and cytochrome c. Future studies of these cybrids will enable us to better understand how Lewy bodies form and what role they play in the pathogenesis of PD. PMID- 14756802 TI - An ALS mouse model with a permeable blood-brain barrier benefits from systemic cyclosporine A treatment. AB - To test potentially beneficial drugs to amyotrophic lateral sclerosis (ALS), we created an ALS mouse model with a permeable blood-brain barrier, by crossing the G93A-SOD1 transgenic mouse with a multiple drug resistance type 1a/b (mdr1a/b) gene knockout mouse. To validate the model, we administered cyclosporine A intraperitoneally to the mice. Cyclosporine A accumulated in the brain and spinal cord of this mouse model, whereas it was unable to penetrate the CNS of mdr1a/b wild-type animals. Systemic administration of cyclosporine A extended the life of the double-mutant male mice by approximately 12%. Surprisingly, the effect was more robust in male mice and only marginal in female mice. These results demonstrate the usefulness of this combined mouse model for the testing of potentially therapeutic drugs and support the role of mitochondrial-mediated apoptosis in the pathway to motor neuron death in SOD1-associated ALS. PMID- 14756801 TI - Brain capillary endothelium and choroid plexus epithelium regulate transport of transferrin-bound and free iron into the rat brain. AB - Iron transport into the CNS is still not completely understood. Using a brain perfusion technique in rats, we have shown a significant brain capillary uptake of circulating transferrin (Tf)-bound and free 59Fe (1 nm) at rates of 136 +/- 26 and 182 +/- 23 microL/g/min, respectively, while their respective transport rates into brain parenchyma were 1.68 +/- 0.56 and 1.52 +/- 0.48 microL/g/min. Regional Tf receptor density (Bmax) in brain endothelium determined with 125I-holo-Tf correlated well with 59Fe-Tf regional brain uptake rates reflecting significant vascular association of iron. Tf-bound and free circulating 59Fe were sequestered by the choroid plexus and transported into the CSF at low rates of 0.17 +/- 0.01 and 0.09 +/- 0.02 microL/min/g, respectively, consistent with a 10-fold brain-CSF concentration gradient for 59Fe, Tf-bound or free. We conclude that transport of circulating Tf-bound and free iron could be equally important for its delivery to the CNS. Moreover, data suggest that entry of Tf-bound and free iron into the CNS is determined by (i) its initial sequestration by brain capillaries and choroid plexus, and (ii) subsequent controlled and slow release from vascular structures into brain interstitial fluid and CSF. PMID- 14756803 TI - Enhanced morphine withdrawal and micro -opioid receptor G-protein coupling in A2A adenosine receptor knockout mice. AB - Much evidence supports the hypothesis that A2A adenosine receptors play an important role in the expression of morphine withdrawal and that the dopaminergic system might also be involved. We have evaluated morphine withdrawal signs in wild-type and A2A receptor knockout mice and shown a significant enhancement in some withdrawal signs in the knockout mice. In addition, micro -opioid and dopamine D2 receptor autoradiography, as well as micro -opioid receptor stimulated guanylyl 5'-[gamma-[35S]thio]-triphosphate ([35S]GTPgammaS) autoradiography was carried out in brain sections of withdrawn wild-type and knockout mice. No significant changes in D2 and micro -opioid receptor binding were observed in any of the brain regions analysed. However, a significant increase in the level of micro receptor-stimulated [35S]GTPgammaS binding was observed in the nucleus accumbens of withdrawn knockout mice. These data indicate that the A2A receptor plays a role in opioid withdrawal related to functional receptor activation. PMID- 14756804 TI - Glutamate regulates Oct-2 DNA-binding activity through alpha-amino-3-hydroxy-5 methylisoxazole-4-propionate receptors in cultured chick Bergmann glia cells. AB - Ionotropic glutamate receptors in cerebellar Bergmann glial cells are linked to transcriptional regulation and, by these means, are thought to play an important role in plasticity, learning and memory and in several neuropathologies. Within the CNS, the transcription factors of the POU family bind their target DNA sequences after a growth factor-dependent phosphorylation-dephosphorylation cascade. Exposure of cultured Bergmann glial cells to glutamate leads to a time- and dose-dependent increase in Oct-2 DNA-binding activity. The use of specific pharmacological tools established the involvement of Ca2+-permeable alpha-amino-3 hydroxy-5-methylisoxazole-4-propionate receptors. Furthermore, the signaling cascade includes phosphatidyl inositol 3-kinase as well as protein kinase C activation. Interestingly, transcriptional as well as translational inhibitors abolish the glutamate effect, suggesting a transcriptional up-regulation of the oct-2 gene. These data demonstrate that Oct-2 expression is not restricted to neurons and further strengthen the notion that the glial glutamate receptors participate in the modulation of glutamatergic cerebellar neurotransmission. PMID- 14756805 TI - Microglia/macrophage-specific protein Iba1 binds to fimbrin and enhances its actin-bundling activity. AB - Ionized calcium binding adaptor molecule 1 (Iba1) is a microglia/macrophage specific calcium-binding protein. Iba1 has the actin-bundling activity and participates in membrane ruffling and phagocytosis in activated microglia. In order to understand the Iba1-related intracellular signalling pathway in greater detail, we employed a yeast two-hybrid screen to isolate an Iba1-interacting molecule and identified another actin-bundling protein, L-fimbrin. In response to stimulation, L-fimbrin accumulated and co-localized with Iba1 in membrane ruffles induced by M-CSF-stimulation and phagocytic cups formed by IgG-opsonized beads in microglial cell line MG5. L-fimbrin was shown to associate with Iba1 in cell lysate of COS-7 expressing L-fimbrin and Iba1. By using purified proteins, direct binding of Iba1 to L-fimbrin was demonstrated by immunoprecipitation, glutathione S-transferase pull-down assays and ligand overlay assays. The binding of Iba1 was also found to increase the actin-bundling activity of L-fimbrin. These results indicate that Iba1 forms complexes with L-fimbrin in membrane ruffles and phagocytic cups, and suggest that Iba1 co-operates with L-fimbrin in modulating actin reorganization to facilitate cell migration and phagocytosis by microglia. PMID- 14756806 TI - Cell type-dependent recruitment of trichostatin A-sensitive repression of the human 5-HT1A receptor gene. AB - Regulation of serotonin (5-HT)1A receptor expression in brain is implicated in mood disorders such as depression and anxiety. Transcriptional activity of the human 5-HT1A receptor gene was strongly repressed by a negative regulatory region containing a consensus repressor element-1 (RE-1) and two copies of the dual repressor element (DRE) identified in the rat 5-HT1A receptor gene. REST/NRSF, a silencer of neuronal genes, bound the 5-HT1A RE-1 and repressed the 5-HT1A promoter. Inactivation of RE-1 completely abolished REST-mediated repression, but resulted in only partial (15-50%) de-repression of basal 5-HT1A promoter activity. The human 5-HT1A DRE sequences bound specifically to the novel repressor Freud-1 (5'repressor element under dual repression binding protein-1) and conferred repressor activity at 5-HT1A or SV40 promoters. In 5-HT1A-negative cells [L6, human embryonic kidney (HEK) 293], the histone deacetylase (HDAC) inhibitor trichostatin A (TSA) abolished repression mediated by both RE-1/REST and DRE/Freud-1, and induced almost complete de-repression of the 5-HT1A gene. By contrast, in 5-HT1A-expressing neuronal cells (RN46A, SN-48) TSA blocked RE 1/REST repression, but did not affect DRE/Freud-1-mediated repression. Thus in contrast to REST, Freud-1 mediates HDAC-independent repression of the 5-HT1A receptor promoter in neuronal 5-HT1A-positive cells, suggesting that HDAC recruitment might influence neuron-specific gene expression by further silencing expression in non-neuronal tissue. PMID- 14756807 TI - Hypoxic remodelling of Ca2+ mobilization in type I cortical astrocytes: involvement of ROS and pro-amyloidogenic APP processing. AB - Chronic hypoxia (CH) alters Ca2+ homeostasis in various cells and may contribute to disturbed Ca2+ homeostasis of Alzheimer's disease. Here, we have employed microfluorimetric measurements of [Ca2+]i to investigate the mechanism underlying augmentation of Ca2+ signalling by chronic hypoxia in type I cortical astrocytes. Application of bradykinin evoked significantly larger rises of [Ca2+]i in hypoxic cells as compared with control cells. This augmentation was prevented fully by either melatonin (150 micro m) or ascorbic acid (200 micro m), indicating the involvement of reactive oxygen species. Given the association between hypoxia and increased production of amyloid beta peptides (AbetaPs) of Alzheimer's disease, we performed immunofluorescence studies to show that hypoxia caused a marked and consistent increased staining for AbetaPs and presenilin-1 (PS-1). Western blot experiments also confirmed that hypoxia increased PS-1 protein levels. Hypoxic increases of AbetaP production was prevented with inhibitors of either gamma- or beta-secretase. These inhibitors also partially prevented the augmentation of Ca2+ signalling in astrocytes. Our results indicate that chronic hypoxia enhances agonist-evoked rises of [Ca2+]i in cortical astrocytes, and that this can be prevented by antioxidants and appears to be associated with increased AbetaP formation. PMID- 14756808 TI - Hypoxia increases calcium flux through cortical neuron glutamate receptors via protein kinase C. AB - The effects of 30 s to 10 min hypoxia (PO2-10 mmHg) on glutamate receptor activity were studied in murine cortical neurons. Receptor activity was assessed as a rise in intracellular calcium concentration ([Ca2+]i) following a 10 s application of 1 mm glutamate or 100 micro mN-methy-d-aspartate (NMDA) in the presence of 0.1 mm Mg2+ and 10 micro m glycine. Change in [Ca2+]i elicited by glutamate increased 26% (n = 192, p < 0.001) and that to NMDA by 74% (n = 9, p < 0.01) during a 100-s period of hypoxia. After 10 min hypoxia, responses to glutamate were 62% smaller than those in normoxia, with increased basal intracellular [Ca2+]i predicting reduced receptor activity. When neurons were exposed to NMDA after 10 min of hypoxia, [Ca2+]i increases were 12% smaller than after 100 s hypoxia, but still 53% larger than in oxygenated neurons (n = 9, p = 0.01). Neurons expressed relatively similar amounts of NR2A, -B, -C, and -D subunits. The phosphorylation of NMDA NR1 subunits increased during hypoxia. Pre treatment of neurons with a protein kinase C (PKC) inhibitor (chelerythrine, 10 micro m) prevented increases in N-methy-d-aspartate receptor (NMDAR) activity during hypoxia and reduced the phosphorylation of NR1 subunits. These results suggest that enhancement of glutamate receptor activity during the first minutes of hypoxia is mediated by phosphorylation of NMDARs by PKC and that other mechanisms, possibly involving intracellular calcium, limit glutamate receptor mediated calcium influx during longer periods of hypoxia. PMID- 14756809 TI - Glycogen metabolism in the rat retina. AB - It has been reported that glycogen levels in retina vary with retinal vascularization. However, the electrical activity of isolated retina depends on glucose supply, suggesting that it does not contain energetic reserves. We determined glycogen levels and pyruvate and lactate production under various conditions in isolated retina. Ex vivo retinas from light- and dark-adapted rats showed values of 44 +/- 0.3 and 19.5 +/- 0.4 nmol glucosyl residues/mg protein, respectively. The glycogen content of retinas from light-adapted animals was reduced by 50% when they were transferred to darkness. Glycogen levels were low in retinas incubated in glucose-free media and increased in the presence of glucose. The highest glycogen values were found in media containing 20 mm of glucose. A rapid increase in lactate production was observed in the presence of glucose. Surprisingly, glycogen levels were the lowest and lactate production was also very low in the presence of 30 mm glucose. Our results suggest that glycogen can be used as an immediate accessible energy reserve in retina. We speculate on the possibility that gluconeogenesis may play a protective role by removal of lactic acid. PMID- 14756810 TI - Expression and involvement of gicerin, a cell adhesion molecule, in the development of chick optic tectum. AB - Gicerin is a cell adhesion molecule belonging to the immunoglobulin superfamily. It has both a homophilic binding activity and a heterophilic binding activity to neurite outgrowth factor (NOF) a molecule belonging to the laminin family. We have reported many studies on the heterophilic activity of gicerin and NOF, but the function of its homophilic binding activity in vivo had been unclear. In the retina, gicerin is expressed in retinal ganglion cells only when they extend neurites to the optic tectum. In this report we have found that gicerin is also transiently expressed in the optic tectum during this time. First, cell aggregation assays were used to show that gicerin expressed in the optic tectum displays homophilic binding activity. Then, explant cultures of embryonic day 6 chick optic tectum on gicerin-Fc chimeric protein-coated dishes and NOF-coated dishes were carried out. It was found that gicerin-gicerin homophilic interactions promoted cell migration, whereas heterophilic interactions with NOF induced neurite formation. Furthermore, when anti-gicerin antibodies were injected in order to examine the effect of gicerin protein in the formation of the tectal layer in ovo, cell migration was strongly inhibited. These data suggest that homophilic interaction of gicerin participates in the migration of neural cells during the layer formation and plays a crucial role in the organization of the optic tectum. PMID- 14756811 TI - Glucocorticoids modulate neurotransmitter-induced glycogen metabolism in cultured cortical astrocytes. AB - Glucocorticoids (GC) are considered as key modulators of glycogen homeostasis in peripheral tissues, but their role in the central nervous system has only partially been characterized. Exposure of primary cultures of cortical astrocytes to dexamethasone (DEX), a synthetic glucocorticoid, results in the reduction of noradrenaline (NA)-induced glycogen synthesis in a concentration-dependent manner with a IC50 of 4.88 nm and a maximum inhibition of 51%. Such an effect is mediated via glucocorticoid receptors (GRs), since it is mimicked by the glucocorticoid analogue RU28362 (100 nm) and prevented by the GR antagonist RU38486 (1 micro m). DEX does not act through alteration of signal transduction mechanisms, as cAMP formation induced by noradrenergic stimulation was unchanged. Moreover, glycogen synthesis was inhibited to the same extent when DEX was applied either together or only after a brief NA application. Neither [3H]2 deoxyglucose uptake nor lactate release was altered by DEX in the presence of NA, demonstrating that inhibition of glycogen synthesis is not a consequence of reduced glucose utilization or availability. Interestingly, enhancement of glycogen synthase activity induced by NA was reduced in the presence of DEX ( 27%). These results suggest that GC could have a significant influence on neuroenergetics as they could modulate activity-related changes in brain glycogen metabolism. PMID- 14756812 TI - Adaptive responses in hypothalamic neuropeptide Y in the face of prolonged high fat feeding in the rat. AB - While a dysregulation in neuropeptide Y (NPY) signaling has been described in rodent models of obesity, few studies have investigated the time-course of changes in NPY content and responsiveness during development of diet-induced obesity. Therefore we investigated the effect of differing lengths (2-17 weeks) of high-fat diet on hypothalamic NPY peptide content, release and NPY-induced hyperphagia. Male Sprague-Dawley rats (211 +/- 3 g) were fed either a high-fat diet (30% fat) or laboratory chow (5% fat). Animals were implanted with intracerebroventricular cannulae to investigate feeding responses to NPY (0.5 nmol, 1 nmol) after 4 or 12 weeks of diet. At the earlier stage of obesity, NPY induced hyperphagia was not altered; however, animals maintained on the high-fat diet for the longer duration were hyper-responsive to NPY, compared to chow-fed control rats (p < 0.05). Overall, hypothalamic NPY peptide content tended to be decreased from 9 to 17 weeks of diet (p < 0.05). Total hypothalamic NPY content was negatively correlated with plasma leptin concentration (p < 0.05), suggesting the hypothalamic NPY system remains responsive to leptin's inhibitory signal. In addition, hypothalamic NPY overflow was significantly reduced in high-fat fed animals (p < 0.05). Together these results suggest a reduction in hypothalamic NPY activity in high-fat fed animals, perhaps in an attempt to restore energy balance. PMID- 14756813 TI - Noradrenaline transporter blockers raise extracellular dopamine in medial prefrontal but not parietal and occipital cortex: differences with mianserin and clozapine. AB - This study compared the interaction between noradrenaline (NA) and dopamine (DA) mechanisms in the prefrontal (PFCX) and in the parietal (ParCX) and occipital (OccCX) cortex. The effect of reboxetine and desipramine, two NA transporter blockers, of mianserin, an antagonist of alpha2 and 5-HT2 receptors, and of clozapine, an atypical antipsychotic, on dialysate DA in the medial PFCX, ParCX and OccCX was studied. We also assessed the influence of a prior 6 hydroxydopamine (6-OHDA) lesion of the dorsal noradrenergic bundle (DNAB) on the effect of reboxetine and clozapine on dialysate DA in the PFCX and ParCX. Systemic administration of reboxetine and desipramine dose-dependently increased dialysate DA in the PFCX but not in the ParCX and OccCX. In contrast, mianserin and clozapine raised dialysate DA in the ParCX and OccCX to an even larger extent than in the PFCX. 6-OHDA lesions of DNAB abolished the increase of dialysate DA elicited by reboxetine in the PFCX and by clozapine both in the PFCX and in the ParCX. It is concluded that, although PFCX and ParCX/OccCX share the presence of a strong control of DA transmission by NA through alpha2 receptors, they differ in the extent to which DA is cleared from the extracellular compartment by uptake through the NA transporter. This process, although extensive in the PFCX, appears insignificant in the ParCX and OccCX, probably as a result of the higher ratio of NA to DA resulting in exclusion of DA from NA transporter. PMID- 14756814 TI - Up-regulation of glutamate concentration in the putamen and in the prefrontal cortex of asymptomatic SIVmac251-infected macaques without major brain involvement. AB - We quantified putamen and prefrontal cortex metabolites in macaques with simian immunodeficiency virus infection and searched for virological and histological correlates. Fourteen asymptomatic macaques infected since 8-78 months (median: 38) were compared with eight uninfected ones. Absolute concentrations of acetate, alanine, aspartate, choline, creatine, GABA, glutamate, glutamine, lactate, myo inositol, N-acetylaspartate, taurine and valine were determined by ex vivo proton magnetic resonance spectroscopy. Glutamate concentration in the CSF was determined by HPLC. Gliosis was assessed by glial fibrillary acidic protein and CD68 immunohistochemistry. Glutamate concentration was slightly increased in the prefrontal cortex (19%, p = 0.0152, t-test) and putamen (13%, p = 0.0354, t-test) of the infected macaques, and was unaffected in the CSF. Myo-inositol concentration was increased in the prefrontal cortex only (27%, p = 0.0136). The concentrations of glutamate and myo-inositol in the prefrontal cortex were higher in the animals with marked or intense microgliosis (p = 0.0114). The other studied metabolites, including N-acetylaspartate, were not altered. Glutamate concentration may thus increase in the cerebral parenchyma in asymptomatic animals, but is not accompanied by a detectable decrease in N-acetylaspartate concentration (neuronal dysfunction). Thus, there are probably compensatory mechanisms that may limit glutamate increase and/or counterbalance its effects. PMID- 14756815 TI - Role of reactive oxygen species in LPS-induced production of prostaglandin E2 in microglia. AB - We determined the roles of reactive oxygen species (ROS) in the expression of cyclooxygenase-2 (COX-2) and the production of prostaglandin E2 (PGE2) in lipopolysaccharide (LPS)-activated microglia. LPS treatment increased intracellular ROS in rat microglia dose-dependently. Pre-treatment with superoxide dismutase (SOD)/catalase, or SOD/catalase mimetics that can scavenge intracellular ROS, significantly attenuated LPS-induced release in PGE2. Diphenylene iodonium (DPI), a non-specific NADPH oxidase inhibitor, decreased LPS induced PGE2 production. In addition, microglia from NADPH oxidase-deficient mice produced less PGE2 than those from wild-type mice following LPS treatment. Furthermore, LPS-stimulated expression of COX-2 (determined by RT-PCR analysis of COX-2 mRNA and western blot for its protein) was significantly reduced by pre treatment with SOD/catalase or SOD/catalase mimetics. SOD/catalase mimetics were more potent than SOD/catalase in reducing COX-2 expression and PGE2 production. As a comparison, scavenging ROS had no effect on LPS-induced nitric oxide production in microglia. These results suggest that ROS play a regulatory role in the expression of COX-2 and the subsequent production of PGE2 during the activation process of microglia. Thus, inhibiting NADPH oxidase activity and subsequent ROS generation in microglia can reduce COX-2 expression and PGE2 production. These findings suggest a potential therapeutic intervention strategy for the treatment of inflammation-mediated neurodegenerative diseases. PMID- 14756816 TI - Protein associated with Myc (PAM) is involved in spinal nociceptive processing. AB - PAM (protein associated with Myc) is a potent inhibitor of adenylyl cyclases (ACs) which is primarily expressed in neurones. Here we describe that PAM is highly expressed in dorsal horn neurones and motoneuron of the spinal cord, as well as in neurones of dorsal root ganglia in adult rats. PAM mRNA expression is differentially regulated during development in both spinal cord and dorsal root ganglia of rats, being strongest during the major respective synaptogenic periods. In adult rats, PAM expression was up-regulated in the spinal cord after peripheral nociceptive stimulation using zymosan and formalin injection, suggesting a role for PAM in spinal nociceptive processing. Since PAM inhibited Galphas-stimulated AC activity in dorsal root ganglia as well as spinal cord lysates, we hypothesized that PAM may reduce spinal nociceptive processing by inhibition of cAMP-dependent signalling. Accordingly, intrathecal treatment with antisense but not sense oligonucleotides against PAM increased basal and Galphas stimulated AC activity in the spinal cord and enhanced formalin-induced nociceptive behaviour in adult rats. Taken together our findings demonstrate that PAM is involved in spinal nociceptive processing. PMID- 14756817 TI - Normal levels of tryptophan hydroxylase immunoreactivity in the dorsal raphe of depressed suicide victims. AB - A variety of evidence suggests that serotonin neurotransmission is altered in the brain of suicide victims and depressed patients. While numerous post-mortem studies have investigated serotonin transporters and receptors, few studies have examined the biosynthetic integrity of the rate-limiting enzyme, tryptophan hydroxylase (TPH), in post-mortem specimens of depressed suicide subjects. Therefore, the aim of the present study was to test the hypothesis that the levels of TPH immunoreactivity (IR) are altered in specific subnuclei of the dorsal raphe (DR) in depressed suicide victims. Suicide victims with a confirmed diagnosis of major depression were matched with non-psychiatric controls based on age, gender and post-mortem interval. Frozen tissue sections containing the DR were selected from two anatomical levels and processed for TPH radioimmunocytochemistry. The optical density corresponding to the regional levels of TPH-IR was quantified in specific subnuclei of the DR from the film autoradiographic images. No significant differences in the levels of TPH-IR were found in any DR subnuclei between depressed suicide victims and control subjects. The lack of change in TPH-IR levels does not necessarily imply that serotonin synthesis or neurotransmission is not altered in the brain of depressed subjects. Many factors influence and regulate serotonin synthesis, and it is conceivable that alterations exist at other levels of regulation of serotonin biosynthesis in depression. Our findings indicate that TPH biosynthesis, at least at the protein level, is not significantly altered in the DR of depressed suicide victims. PMID- 14756818 TI - Neuregulin-1-beta1 enters brain and spinal cord by receptor-mediated transport. AB - Proteins of the neuregulin (NRG) family play important regulatory roles in neuronal survival and synaptic activity. NRG-1-beta1 has particular potential as a therapeutic agent because it enhances myelination of neurites in spinal cord explants. In this study, we determined the permeation of NRG-1-beta1 across the blood-brain and blood-spinal cord barriers (BBB and BSCB respectively). Intact radioactively labeled NRG-1-beta1 had a saturable and relatively rapid influx rate from blood to the CNS in mice. Capillary depletion studies showed that NRG-1 beta1 entered the parenchyma of the brain and spinal cord rather than being trapped in the capillaries that compose the BBB. The possible mechanism of receptor-mediated transport was shown by the ability of antibodies to erbB3 and erbB4 receptors to inhibit the influx. Lipophilicity, less important for such saturable transport mechanisms, was measured by the octanol : buffer partition coefficient and found to be low. The results indicate that NRG-1-beta1 enters spinal cord and brain by a saturable receptor-mediated mechanism, which provides the opportunity for possible therapeutic manipulation at the BBB level. PMID- 14756819 TI - X11alpha impairs gamma- but not beta-cleavage of amyloid precursor protein. AB - The phosphotyrosine binding domain of the neuronal protein X11alpha/mint-1 binds to the C-terminus of amyloid precursor protein (APP) and inhibits catabolism to beta-amyloid (Abeta), but the mechanism of this effect is unclear. Coexpression of X11alpha or its PTB domain with APPswe inhibited secretion of Abeta40 but not APPsbetaswe, suggesting inhibition of gamma- but not beta-secretase. To further probe cleavage(s) inhibited by X11alpha, we coexpressed beta-secretase (BACE-1) or a component of the gamma-secretase complex (PS-1Delta9) with APP, APPswe, or C99, with and without X11alpha, in HEK293 cells. X11alpha suppressed the PS 1Delta9-induced increase in Abeta42 secretion generated from APPswe or C99. However, X11alpha did not impair BACE-1-mediated proteolysis of APP or APPswe to C99. In contrast to impaired gamma-cleavage of APPswe, X11alpha or its PTB domain did not inhibit gamma-cleavage of NotchDeltaE to NICD (the Notch intracellular domain). The X11alpha PDZ-PS.1Delta9 interaction did not affect gamma-cleavage activity. In a cell-free system, X11alpha did not inhibit the catabolism of APP C terminal fragments. These data suggest that X11alpha may inhibit Abeta secretion from APP by impairing its trafficking to sites of active gamma-secretase complexes. By specifically targeting substrate instead of enzyme X11alpha may function as a relatively specific gamma-secretase inhibitor. PMID- 14756820 TI - Brain trauma induces X-box protein 1 processing indicative of activation of the endoplasmic reticulum unfolded protein response. AB - Brain trauma was induced in mice using a closed head injury (CHI) model. At 1, 6 or 24 h after trauma, brains were dissected into the cortex, striatum and hippocampus. Changes in levels of processed X-box protein 1 (xbp1), glucose regulated protein 78 (grp78), growth arrest and DNA damage-inducible gene 153 (gadd153) and heat-shock protein 70 (hsp70) mRNA, indicating impaired endoplasmic reticulum (ER) and cytoplasmic functioning, were evaluated by quantitative PCR. In the cortex, processed xbp1 mRNA levels rose to 2000% of control 1 h after CHI, and stayed high throughout the experiments. In the hippocampus and striatum, processed xbp1 mRNA levels rose in a delayed fashion, peaking at 6 h (1000% of control) and 24 h after CHI (1500% of control) respectively. Levels of grp78 mRNA were only slightly increased in the cortex 24 h after CHI (150% of control), and were unchanged or transiently decreased in the hippocampus and striatum. Levels of gadd153 mRNA did not change significantly after trauma. A transient rise in hsp70 mRNA levels was observed only in the cortex, peaking at 1 h after CHI (600% of control). Processing of xbp1 mRNA is a sign of activation of the unfolded protein response indicative of ER dysfunction. The results suggest that brain trauma induces ER dysfunction, which spreads from the ipsilateral cortex to the hippocampus and striatum. These observations may have clinical implications and should therefore be considered for future investigations on therapeutic intervention of brain injury caused by contusion-induced neurotrauma. PMID- 14756821 TI - Down-regulation of rat brain adenosine A1 receptors at the end of pregnancy. AB - The status of the adenosine A1 receptor/adenylyl cyclase (A1R/AC) transduction pathway in rat brain was analysed at the end of pregnancy using different approaches. Pregnancy at term caused a significant decrease in the Bmax value obtained by saturation binding assays using [3H]DPCPX as radioligand, suggesting a down-regulation of adenosine A1 receptor. Moreover, A1 receptor immunodetection in pregnant rat membranes and the level of mRNA coding A1 receptor were significantly decreased. This loss of A1 receptor was associated with a significant increase in receptor affinity, since the KD value from the [3H]DPCPX saturation curve and Ki for N6-cyclohexyladenosine (CHA) were decreased in pregnant rats. Surprisingly, CHA-mediated inhibition of adenylyl cyclase was increased, reflecting enhanced receptor responsiveness. On the other hand, immunoblotting of different alphaGi-protein isoforms revealed a significant increase in alphaGi3 level in membranes from pregnant rats. Pre-incubation of membranes with anti-alphaGi3 antibody blocked the guanosine triphosphate (GTP) or CHA inhibitory effect on adenylyl cyclase in both pregnant and non-pregnant rats, pointing to alphaGi3 as the main isoform involved in the A1 receptor response. These results suggest that, at the end of pregnancy, there is a down-regulation of adenosine A1 receptors counterbalanced with a strengthened functionality, probably due to an increase in both alphaGi3 protein and receptor affinity. PMID- 14756822 TI - Alpha2-adrenoceptor mediated co-release of dopamine and noradrenaline from noradrenergic neurons in the cerebral cortex. AB - Previous results suggest that extracellular dopamine (DA) in the rat cerebral cortex originates from dopaminergic and noradrenergic terminals. To further clarify this issue, dialysate DA, dihydroxyphenylacetic acid (DOPAC) and noradrenaline (NA) were measured both in the medial prefrontal cortex (mPFC) and in the occipital cortex (OCC), with dense and scarce dopaminergic projections, respectively. Moreover, the effect of the alpha2-adrenoceptor antagonist RS 79948 and the D2-receptor antagonist haloperidol on extracellular DA, DOPAC and NA was investigated. Extracellular DA and DOPAC concentrations in the OCC were 43% and 9%, respectively, those in the mPFC. Haloperidol (0.1 mg/kg i.p.) increased DA and DOPAC (by 35% and 150%, respectively) in the mPFC, but was ineffective in the OCC. In contrast, RS 79948 (1.5 mg/kg i.p.) increased NA, DA and DOPAC, both in the mPFC (by approximately 50%, 60% and 130%, respectively) and the OCC (by approximately 50%, 80% and 200%, respectively). Locally perfused, the DA transporter blocker GBR 12909 (10 micro m) was ineffective in either cortex, whereas desipramine (DMI, 100 micro m) markedly increased extracellular NA and DA in both cortices. The weak haloperidol effect on DA efflux was not enhanced after DA- and NA-transporter blockade, whereas after DMI, RS 79948 markedly increased extracellular NA, and especially DA and DOPAC in both cortices. The results support the hypothesis that most extracellular DA in the cortex is co-released with NA from noradrenergic terminals, such co-release being primarily controlled by alpha2-adrenoceptors. PMID- 14756823 TI - Regulation of beta-amyloid precursor protein expression by brain-derived neurotrophic factor involves activation of both the Ras and phosphatidylinositide 3-kinase signalling pathways. AB - Brain-derived neurotrophic factor (BDNF) stimulates beta-amyloid precursor protein (APP) promoter activity by a Ras-dependent mechanism in TrkB-expressing SH-SY5Y cells. To determine the signalling pathways involved in the BDNF-induced response, we have analysed the ability of TrkB mutated forms to mediate promoter stimulation. Brain-derived neurotrophic factor causes a significant induction of promoter activity and mutation K540R in the active site of TrkB completely abolishes the neurotrophin-induced response. A substitution of the Y484 residue by phenylalanine, which blocks binding of Shc, reduces the activation of APP promoter by BDNF by approximately 50% whereas mutation Y785P, which blocks binding of phospholipase C gamma, does not affect the response. In addition, the phosphatidylinositide 3-kinase (PI3K)-specific inhibitors wortmannin and LY294002 reduced BDNF-induced activation. In agreement with a participation of both Ras/MAPK- and PI3K/Akt-mediated mechanisms, transient expression of constitutive active forms of Ras, PI3K and other components of both signalling pathways led to a significant increase of APP promoter activity. Furthermore, the stimulation of the APP promoter by BDNF was completely precluded by expression of dominant negative forms of Ras and PI3K effectors. Taken together, our results suggest that simultaneous activation of at least two signalling pathways, Ras/MAPK and PI3K/Akt, is necessary to mediate a full activation of the APP promoter by BDNF. PMID- 14756824 TI - Overexpression of torsinA in PC12 cells protects against toxicity. AB - Childhood-onset dystonia is an autosomal dominant movement disorder associated with a three base pair (GAG) deletion mutation in the DYT1 gene. This gene encodes a novel ATP-binding protein called torsinA, which in the central nervous system is expressed exclusively in neurons. Neither the function of torsinA nor its role in the pathophysiology of DYT1 dystonia is known. In order to better understand the cellular functions of torsinA, we established PC12 cell lines overexpressing wild-type or mutant torsinA and subjected them to various conditions deleterious to cell survival. Treatment of control PC12 cells with an inhibitor of proteasomal activity, an oxidizing agent, or trophic withdrawal, resulted in cell death, whereas PC12 cells that overexpressed torsinA were significantly protected against each of these treatments. Overexpression of mutant torsinA failed to protect cells against trophic withdrawal. These results suggest that torsinA may play a protective role in neurons against a variety of cellular insults. PMID- 14756827 TI - We need more nursing ethics research. PMID- 14756825 TI - Sphingosine-1-phosphate induces proliferation and morphological changes of neural progenitor cells. AB - Sphingosine-1-phosphate (S1P) is a lipid mediator that exerts multiple cellular functions through activation of G-protein-coupled receptors. Although the role of S1P on angiogenesis is well established, its role in neurogenesis is unknown. We examined the effects of S1P on G-protein activation in brain sections of rat embryo and on neural progenitor cells in culture. Intense S1P-stimulated [35S]GTPgammaS labeling was observed as early as E15 in the neuroepithelium and differentiating fields throughout the brain, suggesting that functional S1P receptors are expressed in brain areas with active neurogenesis. mRNA transcripts for several S1P receptor subtypes (S1P1, S1P2, S1P3 and S1P5) were expressed in neural progenitor cells prepared from embryonic rat hippocampus. S1P induced phosphorylation of extracellular signal-regulated kinase (ERK) and proliferation of neural progenitor cells as determined by BrdU incorporation in a pertussis toxin-sensitive manner. These effects were prevented by the ERK signaling inhibitor U0126. S1P augmented telomerase activity in neural progenitor cells with similar potency as that of FGF-2. Furthermore, S1P induced cell-cell aggregation. This morphological change was transient and prevented by Y-27632, an inhibitor of Rho-associated kinase. These results suggest that S1P plays a pleiotropic role in neurogenesis via pathways involving S1P receptors, MAP kinases and Rho kinase. PMID- 14756829 TI - Determining optimal nursing intensity: the RAFAELA method. AB - BACKGROUND: RAFAELA is a modern system of patient classification. In the last few years the system has become widely used in Finland and has aroused international interest. It comprises three parts: (1) The Oulu Patient Classification (OPC) instrument and (2) a file on nurse resources. Using these, the daily nursing care intensity, expressed as OPC points per nurse, can be calculated. The existing nursing care intensity can then be compared with the optimal by using the third instrument, (3) the Professional Assessment of Optimal Nursing Care Intensity Level (PAONCIL). This is a daily questionnaire that nurses complete in a 2-month period at intervals every few years. The daily workload is scored from -3 to +3, where zero is the optimal level. The optimal nursing care intensity per nurse is then defined by using linear regression analysis. No expensive time studies are needed. AIMS: This paper reports on a study which aimed to identify the minimum requirements for determining optimal nursing care intensity that allow the results to be accepted as correct, in terms of: length of the PAONCIL examination period, PAONCIL questionnaire response rate, explanatory power of the regression analysis and mean values of the OPC and PAONCIL instruments. DESIGN: The results of analyses of optimal nursing care intensity from 61 wards in eight Finnish hospitals for the period 1997-2001 are presented. The data do not contain any information about the identity of the patients. METHODS: Linear regression analysis, one-way analysis of variance, t-tests and correlation analysis were used, as well as parameters of distribution of the data. RESULTS: The results of the analysis of optimal nursing care intensity can be regarded as reliable if the PAONCIL response rate is above 70%, the period of examination is at least 3-4 weeks, the mean PAONCIL value is below 0.65 and the explanatory power is above 25%. CONCLUSION: On the basis of the RAFAELA system, the optimal nursing care intensity of a ward can be reliably determined. The prerequisites for achieving reliable results were clear and mostly fulfilled. Because a study period shorter than that which has previously been the practice is enough, the use of this system will be easier than before. The credibility and usefulness of the RAFAELA system have thus received considerable additional confirmation. PMID- 14756830 TI - Occupational changes in nursing: the situation of enrolled nurses. AB - BACKGROUND: The move to one level of qualified nurse in the United Kingdom (UK) is, in part, a consequence of professionalizing strategies. Registered nurses now undertake technical work previously performed by doctors. The role of enrolled nurses, and their career intentions, have not been considered in light of these changes, despite the fact that many still work in the National Health Service. AIM: This paper considers the pursuit of professionalization by nurses, illustrating the argument with findings from an empirical study of conversion to registered nurse by enrolled nurses. METHODS: The paper is based on a secondary analysis of a large data set, originally used to explore ethnic inequalities in nursing. Data from 2968 respondents were analysed to answer a number of research questions relating to the characteristics of different groups of enrolled nurses and predictors of conversion to registered nurse. These included demographic characteristics, markers of career orientation, career progression and job satisfaction. STUDY LIMITATIONS: This study used secondary analysis of data and, therefore, exploration of issues was limited, not least because the data were 10 years old. Also, the design was cross-sectional and respondents' experiences related to different stages of the phenomenon under study and the same group was not studied over time. FINDINGS: Enrolled nurses who had converted to registered nurse were more likely than those who had no intention of converting to: be male, be younger, have been nursing longer, not be working on elderly care wards, have a high career orientation, not have taken a career break, and work full-time. Most of these factors predicted likelihood to convert. Although nurses who converted to registered nurse were more likely to anticipate career progression, they were less likely to be satisfied with their work. CONCLUSION: In attempts to define rewarding nursing work, the importance of ensuring that qualified nurses continue to retain basic nursing care skills should be acknowledged. This may offer an uncomfortable view to those seeking to further the professional status of nursing. PMID- 14756831 TI - Enhancing critical thinking in the preceptorship experience in nursing education. AB - BACKGROUND: To date no research has been conducted to examine the process used in the preceptorship experience to enhance critical thinking at the graduate level. This study provides data that could revitalize the preceptor/learner relationship and in turn influence future preceptorship programmes in graduate nursing education. AIM: The purpose of this study was to examine the preceptorship experience and its role in the enhancement of critical thinking in graduate nursing education. DESIGN: A grounded theory approach was employed, through semi structured, tape-recorded interviews. Overall, 45 interviews were completed with graduate nursing students ranging in age from 26 to 53 years, and master's and doctorally prepared preceptors who ranged in age from 47 to 58 years. In addition, a journal of personal reflections was kept by each of the researchers. Analysis of data included the process of open coding, theoretical coding, selective coding, reduction and comparison. FINDINGS: Data revealed that a process, designated The Relational Process, occurred in the preceptorship experience to enhance the critical thinking ability of graduate nursing students. This process was found to be a complex, ongoing interpersonal dynamic between the graduate student and assigned preceptor. CONCLUSIONS: The relational process that emerged from this study indicates that specific preceptor behaviours are pivotal to the enhancement of critical thinking of graduate nursing students and ultimately impact on the success or failure of the preceptorship experience. PMID- 14756832 TI - Promoting cognitive and metacognitive reflective reasoning skills in nursing practice: self-regulated learning theory. AB - BACKGROUND: Effective clinical reasoning in nursing practice depends on the development of both cognitive and metacognitive skills. While a number of strategies have been implemented and tested to promote these skills, educators have not been able consistently to predict their development. Self-regulated learning theory suggests that this development requires concurrent attention to both the cognitive and metacognitive dimensions of reasoning in nursing care contexts. AIMS: This paper reports on a study to explore the impact of self regulated learning theory on reflective practice in nursing, and to advance the idea that both cognitive and metacognitive skills support the development of clinical reasoning skills. METHODS: Integrative review of published literature in social science, educational psychology, nursing education, and professional education using the Cumulative Index to Nursing and Allied Health (CINAHL), Educational Resource Information Center (ERIC), and American Psychological Association (PsychInfo) Databases. The search included all English language articles with the key words clinical reasoning, cognition, critical thinking, metacognition, reflection, reflective practice, self-regulation and thinking. FINDINGS: Reflective clinical reasoning in nursing practice depends on the development of both cognitive and metacognitive skill acquisition. This skill acquisition is best accomplished through teaching-learning attention to self regulation learning theory. A critical analysis of the literature in the areas of critical thinking and reflective practice are described as a background for contemporary work with self-regulated learning theory. It is apparent that single minded attention to critical thinking, without attention to the influence of metacognition or reflection, is but one perspective on clinical reasoning development. Likewise, single-minded attention to metacognition or reflection, without attention to the influence of critical thinking, is another perspective on clinical reasoning development. While strategies to facilitate critical thinking and reflective practice have been used in isolation from each other, there is evidence to suggest that they are inextricably linked and come together with the use of self-regulated learning prompts. CONCLUSIONS: Students and practising nurses are able to improve their cognitive and metacognitive skills in clinical contexts by using self-regulated learning strategies. The self-regulated learning model in nursing is offered to support teaching and learning of reflective clinical reasoning in nursing practice contexts. PMID- 14756833 TI - Nurses' perceptions of their pharmacology educational needs. AB - BACKGROUND: Pharmacology education in nursing has become increasingly important as nurses' roles in administering, prescribing and educating patients about their medications have grown. Some authors have expressed concern at the lack of science teaching in nurse education, and others have suggested that there is a theory-practice gap in this area of the curriculum. AIM: This paper reports a study to explore nurses' pharmacology education needs by identifying nursing roles that require pharmacology knowledge, and nurses' preparation for practice from preregistration pharmacology education. METHOD: A qualitative approach was used to collect data from a purposive sample of 10 qualified nurses from an emergency admissions unit in a city in the north of England. Semi-structured interviews were transcribed verbatim and categorized using Burnard's 14 stages. FINDINGS: This study revealed a limited understanding of the subject, and dissatisfaction with the teaching of pharmacology, with resulting anxiety on qualifying. Nursing roles identified as requiring pharmacology knowledge included drug administration, patient assessment, nurse prescribing, and patient medication education. CONCLUSION: The findings suggest that, although nurses have a limited understanding of pharmacology, they recognize the need for pharmacology knowledge in practice. Improved pharmacology teaching might increase nurses' confidence in performing drug administration, patient education, and nurse prescribing, and decrease anxieties related to these roles. PMID- 14756834 TI - A typology of knowledge for district nursing assessment practice. AB - BACKGROUND: This paper reports on part of a larger study, the aim of which was to explore the nature of knowledge required by district nurses (DNs) to carry out first assessment visits, and the relationship of this to the decisions they make. Assessment of need is a key and complex component of the DN role. To date, there has been limited exploration of the knowledge underpinning needs assessment in district nursing practice. AIM: The aim of this paper is to identify and categorize the knowledge in use by DNs undertaking first assessment visits, by presenting it in the form of the typology of knowledge which emerged from the study. METHODS: The study used a qualitative, ethnographic design. Eleven DNs were observed undertaking first assessment visits and interviewed twice: following the observed visit and 1 year later, after preliminary data analysis had been undertaken. Approaches to analysis were data driven, and constructing the typology involved uncovering and linking what the DN was seeing, asking and doing. Data were collected during 1997 and 1998. FINDINGS: The findings revealed a breadth and depth of community nursing knowledge that seemed to incorporate an amalgam of theoretical (knowing that) and practice-based (knowing how) knowledge. The findings depict the range and scope of the knowledge in use by the DNs, and challenge the utility of theoretical models that remove knowledge from the context in which it is used and applied. CONCLUSION: The findings presented here provide fresh insight into the 'know-how' of district nursing assessment practice. Whilst the typology requires further testing and refinement in order to enhance understanding of practice, it conceptualizes aspects of district nursing assessment knowledge and addresses the current lack of underpinning principles in district nursing practice. PMID- 14756835 TI - Cancer-related pain in palliative care: patients' perceptions of pain management. AB - BACKGROUND: Pain is still a significant problem for many patients with cancer, despite numerous, clear and concise guidelines for the treatment of cancer related pain. The impact of pain cognition on patients' experiences of cancer related pain remains relatively unexplored. AIM: The aim of this study was to describe how patients with cancer-related pain in palliative care perceive the management of their pain. METHOD: Thirty patients were strategically selected for interviews with open-ended questions, designed to explore the pain and pain management related to their cancer. The interviews were analysed using a phenomenographic approach. FINDINGS: Patients described 10 different perceptions of pain and pain management summarized in the three categories: communication, planning and trust. In terms of communication, patients expressed a need for an open and honest dialogue with health care professionals about all problems concerning pain. Patients expressed an urgent need for planning of their pain treatment including all caring activities around them. When they felt trust in the health care organization as a whole, and in nurses and physicians in particular, they described improved ability and willingness to participate in pain management. While the findings are limited to patients in palliative care, questions are raised about others with cancer-related pain without access to a palliative care team. CONCLUSION: The opportunity for patients to discuss pain and its treatment seems to have occurred late in the course of disease, mostly not until coming in contact with a palliative care team. They expressed a wish to be pain-free, or attain as much pain relief as possible, with as few side effects as possible. PMID- 14756836 TI - Complexity in caring for patients with advanced cancer. AB - BACKGROUND: The gap between nursing research and practice is readily acknowledged in literature, with a variety of strategies suggested for reducing this gap. It is necessary not only to address problems of research implementation in practice, but also to find strategies that strengthen the influence of practitioners on research agendas through more collaborative relationships in order to have an impact on care of patients. A multi-centre research project was therefore initiated by two universities and three health care facilities, aiming to improve quality of care for patients with advanced cancer through a knowledge-exchange programme between nurse researchers, practitioners and students. AIM: The aim of this article is to explore how clinical staff reason about care provision for patients with advanced cancer, through analysis of 20 focus group discussions conducted with staff in three different health care facilities in two Swedish cities. An initial analysis based on grounded theory was complemented with consideration of the interactive process in the focus group discussions, and carried out by a team consisting of senior nurse researchers, clinical experts and nursing instructors. FINDINGS: The findings of the focus group discussions emphasize the complexity of caregiving for patients with advanced cancer. The tension between caregiving ideals and limits imposed by the realities of caregiving in today's health system were striking. Practitioners discussed the organization of care, different constellations of relationships between patients, family members and professionals, and theoretical and experiential knowledge as equally important aspects in dealing with all concrete situations in daily practice. The importance of reflective practice, use of self and ethical reasoning also permeated the focus group discussions. CONCLUSIONS: These findings highlight an integrated need both to influence organizational structures and working relationships, along with increasing knowledge, if sustainable change is to be effected. PMID- 14756837 TI - Assessment of malnutrition in mental health clients: nurses' judgement vs. a nutrition risk tool. AB - BACKGROUND: The existence of malnutrition in general hospitals is well documented. Psychiatric patients are known to have increased risk of malnutrition, yet physical examinations and nutritional assessments rarely take place in psychiatric hospitals. AIM: The purpose of this study was to adapt an established nutrition risk score for use with psychiatric patients, using criteria previously agreed by the care team, and to assess whether the clinical judgement of ward staff alone identified a similar group of patients to be at risk. METHOD: The risk score assessment was compared with a subjective risk assessment made by nursing staff as patients were admitted to the unit. Data were collected for 112 patients. RESULTS: The comparison revealed that nurses did not identify malnutrition in the same patients as the risk score, overlooking 27 (29%) at risk patients. Nurses associated malnutrition with psychotic illness, suggesting that depressed patients are more likely to be overlooked. STUDY LIMITATIONS: Although the risk score was based on a validated tool and its content and face validity were established, it has not itself been validated against criteria of nutritional status (malnutrition). CONCLUSIONS: Implementing routine nutritional screening on such units would assist in identifying at risk patients, enabling referral for dietetic intervention to be made. Providing nutrition education for staff might help to improve knowledge and awareness of malnutrition for this patient group. PMID- 14756840 TI - Formaldehyde as a basis for residential ventilation rates. AB - Traditionally, houses in the US have been ventilated by passive infiltration in combination with active window opening. However in recent years, the construction quality of residential building envelopes has been improved to reduce infiltration, and the use of windows for ventilation also may have decreased due to a number of factors. Thus, there has been increased interest in engineered ventilation systems for residences. The amount of ventilation provided by an engineered system should be set to protect occupants from unhealthy or objectionable exposures to indoor pollutants, while minimizing energy costs for conditioning incoming air. Determining the correct ventilation rate is a complex task, as there are numerous pollutants of potential concern, each having poorly characterized emission rates, and poorly defined acceptable levels of exposure. One ubiquitous pollutant in residences is formaldehyde. The sources of formaldehyde in new houses are reasonably understood, and there is a large body of literature on human health effects. This report examines the use of formaldehyde as a means of determining ventilation rates and uses existing data on emission rates of formaldehyde in new houses to derive recommended levels. Based on current, widely accepted concentration guidelines for formaldehyde, the minimum and guideline ventilation rates for most new houses are 0.28 and 0.5 air changes per hour, respectively. PMID- 14756838 TI - Successful adherence after multiple HIV treatment failures. AB - BACKGROUND: Non-adherence to human immunodeficiency virus (HIV) medications often results in irreparable drug resistance and poor outcomes. Hence, care providers generally think that treatment of HIV disease should be delayed until a person is 'ready' to adhere. However, little research has focused on understanding the process that results in readiness for successful adherence. AIM: The aim of this phenomenological study was to describe and understand the experience and decision making processes of people who became adherent to their HIV medication regimens after previously failing treatment because of non-adherence. METHOD: A Husserlian phenomenological approach was taken, and in-depth interviews were analysed using Giorgi's method of phenomenological description and analysis. FINDINGS: Thirteen HIV-positive men and women who had previously failed two or more treatment regimens because of non-adherence were purposefully selected from two infectious diseases clinics in the Midwest region of the United States. They had achieved and sustained adherence to their HIV medications for 1 year or longer without formal intervention. All participants experienced a 'trigger' event preceding the process that led to the ability to incorporate lifestyle and health behaviour changes necessary for successful adherence. Factors associated with the process leading to adherence were: changing attitudes towards HIV medication, finding the right health care provider, creating the right support system, getting control of life and having goals. CONCLUSIONS: This study demonstrated that HIV-positive individuals who had been non-adherent and had been viewed as 'difficult to treat' nonetheless successfully adhered to treatment once they became 'ready'. Findings from this study implicate that readiness may be a necessary component for successful adherence, particularly in HIV-positive individuals who have previously failed treatment. Understanding the relationship between the phenomenon of readiness and subsequent HIV treatment adherence has implications for clinical decision-making and for development of interventions that enhance adherence and prevent HIV drug resistance. PMID- 14756841 TI - Objective recognition of cough sound as biomarker for aerial pollutants. AB - A relationship among air quality, respiratory health, and comfort in man and animal is widely shown. In general, a state of respiratory discomfort is prevailed by an increase in acoustic audible symptoms. The general concept of sound analysis as an objective contactless non-invasive biomarker for aerial pollution is studied on free-field cough sound of 12 Belgian Landrace piglets. A citric-acid-induced cough sound recognition algorithm with recognition rate of 95% is applied to cough sounds registered in the presence of distinct types of aerial pollutants: irritating gas (ammonia), respirable particles (dust), and temperature. The recognition performance for all aerial pollutants was >90% and maintained 94% on average. It is concluded that sound analysis allows an effective biomarker for all three types of aerial pollution. The generality of the biomarker is hypothesized to be due to the common mechanism involved in protective cough. As a consequence, it is suggested to use sound analysis as a biomarker for respiratory state in studies of exposure to air pollutants. PMID- 14756842 TI - Class separation of buildings with high and low prevalence of SBS by principal component analysis. AB - In this study, we were able to separate buildings with high and low prevalence of sick building syndrome (SBS) using principal component analysis. The prevalence of SBS was defined by the presence of at least one typical skin, mucosal and general (headache and fatigue) symptom. Data from the Swedish Office Illness Study describing the presence and level of chemical compounds in outdoor, supply, and room air, respectively, were evaluated together with information about the buildings in six models. When all data were included the most complex model was able to separate 71% of the high prevalence buildings from the low prevalence buildings. The most important variable that separates the high prevalence buildings from the low prevalence buildings was a more frequent occurrence or a higher concentration of compounds with shorter retention time in the high prevalence buildings. Elevated relative humidity in supply and room air and higher levels of total volatile organic compounds in outdoor and supply air were more common in high prevalence buildings. Ten building variables also contributed to the separation of the two classes of low and high prevalence buildings. PMID- 14756843 TI - The Stockholm Indoor Environment Questionnaire: a sociologically based tool for the assessment of indoor environment and health in dwellings. AB - The aim was to develop and validate a standardized questionnaire - the Stockholm Indoor Environment Questionnaire (SIEQ). The validation procedure was based on sociological principles and test procedures for validation. The indicators of indoor environment are air quality, thermal climate, noise, and illumination. The indicators of health are symptoms comprised in the sick building syndrome (SBS). The questionnaire also contains questions about the apartment, individual behavior, and personal factors. The everyday language describing the building and its function was first obtained by qualitative personal interviews, then by standardized questions. The interview questionnaire was transformed into a postal self-administered questionnaire. The reduction of the questionnaire was based on correlation analysis. It was found that to obtain a good validity, general questions are not sufficient, but specific question on perceptions and observations are needed. Good test-retest agreement was found both on an area level, building level, and individually. For each indicator, a set of questions are constructed and validated. SIEQ has been used in several studies, and the results are presented in graphic problem profiles. Reference data has been calculated for the Stockholm area. PMID- 14756844 TI - Building characteristics, indoor air quality and recurrent wheezing in very young children (BAMSE). AB - This study was conducted to examine the impact of building characteristics and indoor air quality on recurrent wheezing in infants. We followed a birth cohort (BAMSE) comprising 4089 children, born in predefined areas of Stockholm, during their first 2 years of life. Information on exposures was obtained from parental questionnaires when the children were 2 months and on symptoms and diseases when the children were 1 and 2 years old. Children with recurrent wheezing, and two age-matched controls per case, were identified and enrolled in a nested case control study. The homes were investigated and ventilation rate, humidity, temperature and NO2 measured. We found that living in an apartment erected after 1939, or in a private home with crawl space/concrete slab foundation were associated with an increased risk of recurrent wheezing, odds ratio (OR) 2.5 (1.3 4.8) and 2.5 (1.1-5.4), respectively. The same was true for living in homes with absolute indoor humidity >5.8 g/kg, OR 1.7 (1.0-2.9) and in homes where windowpane condensation was consistently reported over several years, OR 2.2 (1.1 4.5). However, air change rate and type of ventilation system did not seem to affect the risk. In conclusion, relatively new apartment buildings, single-family homes with crawl space/concrete slab foundation, elevated indoor humidity, and reported wintertime windowpane condensation were associated with recurrent wheezing in infants. Thus, improvements of the building quality may have potential to prevent infant wheezing. PMID- 14756845 TI - Experimental and CFD evidence of multiple solutions in a naturally ventilated building. AB - This paper considers the existence of multiple solutions to natural ventilation of a simple one-zone building, driven by combined thermal and opposing wind forces. The present analysis is an extension of an earlier analytical study of natural ventilation in a fully mixed building, and includes the effect of thermal stratification. Both computational and experimental investigations were carried out in parallel with an analytical investigation. When flow is dominated by thermal buoyancy, it was found experimentally that there is thermal stratification. When the flow is wind-dominated, the room is fully mixed. Results from all three methods have shown that the hysteresis phenomena exist. Under certain conditions, two different stable steady-state solutions are found to exist by all three methods for the same set of parameters. As shown by both the computational fluid dynamics (CFD) and experimental results, one of the solutions can shift to another when there is a sufficient perturbation. These results have probably provided the strongest evidence so far for the conclusion that multiple states exist in natural ventilation of simple buildings. Different initial conditions in the CFD simulations led to different solutions, suggesting that caution must be taken when adopting the commonly used 'zero initialization'. PMID- 14756846 TI - Microbial contamination of indoor air due to leakages from crawl space: a field study. AB - Mechanical exhaust ventilation system is typical in apartment buildings in Finland. In most buildings the base floor between the first floor apartments and crawl space is not air tight. As the apartments have lower pressure than the crawl space due to ventilation, contaminated air may flow from the crawl space to the apartments. The object of this study was to find out whether a potential air flow from crawl space has an influence on the indoor air quality. The results show that in most cases the concentration of fungal spores was clearly higher in the crawl space than inside the building. The size distribution of fungal spores depended on the fungal species. Correlation between the fungal spores in the crawl space and indoors varied with microbial species. Some species have sources inside the building, which confounds the possible relation between crawl pace and indoor concentrations. Some species, such as Acremonium, do not normally have a source indoors, but its concentration in the crawl space was elevated; our measurements showed also elevated concentrations of Acremonium in the air of the apartments. This consistent finding shows a clear linkage between fungal spores in the indoor air and crawl space. We conclude that a building with a crawl space and pressure difference over the base floor could be a potential risk for indoor air quality in the first floor apartments. PMID- 14756847 TI - Molds in floor dust and building-related symptoms in adolescent school children. AB - This stratified cross-sectional epidemiological study included 1053 school children aged 13-17 years. All pupils filled in a questionnaire on building related symptoms and other relevant health aspects. The following exposure measurements were carried out: room temperature, CO2 level, and relative humidity; building characteristics including mold infestation were assessed, and dust was collected from floors, air, and ventilation ducts during a working day. Dust was examined for endotoxin level, and cultivated for viable molds. We did not find a positive association between building-related symptoms and extent of moisture and mold growth in the school buildings. Five of eight building-related symptoms were significantly and positively associated with the concentration of colony forming units of molds in floor dust: eye irritation, throat irritation, headache, concentration problems, and dizziness. After adjusting for different potentially confounding factors in separate analyses of each symptom, the above mentioned associations between molds in dust and symptoms were still present, except for concentration problems. However, in none of the analyses was mold exposure the strongest covariate, being secondary to either asthma, hay fever, recent airway infection, or psychosocial factors. PMID- 14756849 TI - Efficacy and tolerability of oral zolmitriptan in menstrually associated migraine: a randomized, prospective, parallel-group, double-blind, placebo controlled study. AB - BACKGROUND: Approximately 60% of female migraineurs report experiencing migraine in association with menstruation, while 7% to 25% experience attacks almost exclusively with menstruation. OBJECTIVE: To examine the efficacy and tolerability of oral zolmitriptan in the acute treatment of menstrually associated migraine. In this study, menstrually associated migraine was defined as migraine that consistently occurred from 72 hours before to 5 days after onset of menses. Methods.-Participants were women with regular menstrual cycles, aged 18 to 55 years, who had experienced migraine with at least two thirds of prior menstrual cycles. Subjects were randomized to treat one attack per menstrual cycle for 3 months with either zolmitriptan or placebo. Treatment was intensity based: mild migraines were treated with half of a 2.5-mg zolmitriptan tablet, moderate migraines were treated with zolmitriptan 2.5 mg, and severe migraines were treated with 5 mg (two 2.5-mg tablets) of zolmitriptan, or placebo. RESULTS: Of the 579 women enrolled in the study, 260 were treated with zolmitriptan and 251 were assigned placebo. Twelve hundred thirty-two attacks were treated, and a 2-hour headache response was achieved in 48% of zolmitriptan-treated attacks as compared with 27% of placebo-assigned attacks (P <.0001). Zolmitriptan was superior to placebo in achieving a headache response as early as 30 minutes (18% versus 14%, P=.03) and at 1 hour (33% versus 23%, P <.001). Drug-related adverse events were reported in 16% of subjects receiving zolmitriptan and 9% of subjects receiving placebo. CONCLUSION: Oral zolmitriptan exhibits efficacy and good tolerability in the treatment of menstrually associated migraine. Improvement over placebo was observed as early as 30 minutes following treatment. PMID- 14756850 TI - Consecutive transcranial magnetic stimulation: phosphene thresholds in migraineurs and controls. AB - OBJECTIVE: To characterize the temporal course of transcranial magnetic stimulation-induced phosphene thresholds in subjects with migraine and in controls. METHODS: Eleven subjects with migraine with aura, 10 subjects with migraine without aura, 9 subjects with menstrual migraine, and 15 controls (no history of migraine and without migraine during the study) were studied. Subjects were not on preventive medication. Transcranial magnetic stimulation was performed, and a phosphene threshold was measured 3 times a week over 3 weeks in a manner timed to incorporate the menstrual period in females. A headache calendar was kept during the study. RESULTS: Mean transcranial magnetic stimulation thresholds were lower for each migraine group compared with controls (P <.001) for each comparison. There was a trend for lower thresholds among subjects with migraine with aura compared with subjects with migraine without aura (P <.10), but not subjects with menstrual migraine. There was consistent lowering of thresholds from the first to the last stimulation in all migraine groups and in the controls. Maximum and minimum thresholds did not predict headache occurrence, nor did the occurrence of headache predict an ensuing maximum or minimum phosphene threshold. CONCLUSIONS: Transcranial magnetic stimulation thresholds are lower in subjects with migraine compared with controls. The reported phosphene threshold is lowered with repeated measurement. Neither high nor low phosphene thresholds predict a subsequent headache, nor do migraines predict a subsequent high or low threshold. PMID- 14756851 TI - Efficacy of diclofenac sodium softgel 100 mg with or without caffeine 100 mg in migraine without aura: a randomized, double-blind, crossover study. AB - OBJECTIVE: A phase II, randomized, double-blind, crossover study was designed to evaluate the efficacy of 100-mg diclofenac sodium softgel (formulated using ProSorb technology) with or without 100-mg caffeine versus placebo in migraineurs during migraine attacks. BACKGROUND: Diclofenac has been demonstrated to be an effective migraine treatment in several placebo-controlled studies. A rapidly absorbed softgel of diclofenac has been shown to be effective in the rapid relief of acute pain, and may have advantages in migraine treatment. In addition, caffeine has consistently been shown to increase both the efficacy and speed of onset of concurrently administered analgesics. The ability of caffeine to both enhance and accelerate analgesic effects has been documented with a variety of different medications (ie, aspirin, acetaminophen, ibuprofen, and ergotamine). METHODS: The 3-period crossover study was designed to compare diclofenac softgel 100 mg, diclofenac softgel 100 mg plus caffeine 100 mg, and placebo in the acute treatment of migraine. Subjects treated one moderate or severe attack with each study medication. The primary efficacy parameter was the percentage of subjects with headache relief at 60 minutes as defined by a reduction of headache severity from moderate or severe at baseline to absent or mild compared with placebo. Though the sample size estimate required that 72 subjects treat 3 separate attacks, 51 subjects treated 1 migraine attack, 44 treated 2 attacks, and 39 treated 3 attacks. Results.-In the placebo group, 6 (14%) of 43 subjects reported headache relief at 60 minutes versus 12 (27%) of 45 subjects in the diclofenac softgel group, and 19 (41%) of 46 subjects in the diclofenac softgel plus caffeine group. Differences were statistically significant for the diclofenac softgel plus caffeine group versus placebo (odds ratio, 4.2; 95% confidence interval, 1.3 to 13.7). Rescue medication was used by 27 (63%) of 43 subjects treated with placebo, 15 (33%) of 45 subjects treated with diclofenac softgel, and 14 (30%) of 46 subjects treated with diclofenac softgel plus caffeine. This result is highly statistically significant (chi22= 11.56, P=.003). Both the diclofenac plus caffeine (P <.03) and diclofenac only (P <.03) groups were significantly different from the placebo group in terms of the visual analog scale score at 60 minutes. CONCLUSIONS: The major finding of the present study is that diclofenac softgel plus caffeine produces statistically significant benefits relative to placebo at 60 minutes. Diclofenac softgel alone did not differ significantly from placebo, perhaps due to limits in sample size. Nonsignificant trends support the analgesic adjuvant benefit of caffeine when added to diclofenac softgels. PMID- 14756852 TI - Onset of effect of 5-HT1B/1D agonists: a model with pharmacokinetic validation. AB - OBJECTIVE: To quantitate onset of effect of all formulations of sumatriptan, and to investigate whether this is related to rate, not extent, of drug absorption. METHODS: From published literature, for 4 formulations of sumatriptan and matching placebos, response rates were modeled using a simple logarithmic equation, with a being a parameter of curve convexity and B, a location parameter (equal to response rate at 2 hours [the standard regulatory parameter]). The average rate of drug absorption (A) was estimated by dividing the maximal drug concentration by the time needed to achieve it (Cmax/Tmax). Least mean square correlation was then performed between the therapeutic gains and therapeutic ratios of curve convexity and rate of drug absorption. RESULTS: -Models closely fitted observed response rates (2 hours or less). Curve convexity correlated with rate of drug absorption. Sumatriptan response rates (0 to 2 hours) for formulations correlated with rate, not extent, of drug absorption. The range of rates of onset of effect among different routes of administration was greater than that for tablets with 4-fold differences in dose size. CONCLUSION: Onset of effect is related to rate of absorption of sumatriptan. There is greater scope for improving onset of effect using an alternative route of administration than by increasing the oral dose. PMID- 14756853 TI - Headache induced by chronic substance use: analysis of medication overused and minimum dose required to induce headache. AB - BACKGROUND: The International Headache Society has defined the diagnostic criteria for headache induced by substance use. Recently, a revision to these criteria has been proposed. OBJECTIVE: To consider whether the International Headache Society criteria for headache induced by substance use and the proposed revisions for the classification of daily and near-daily headache with medication abuse permit classification of patients commonly seen in a headache center. METHODS: One hundred fourteen consecutive patients (96 women [84.2%] and 18 men [15.8%]; mean age, 54.2 years [SD, 14]) with headache and chronic overuse of medications, admitted for detoxification to the inpatient unit of a headache center, participated in the study. The initial headache, medications and doses used, duration of daily medication use, and means of medication administration were studied. RESULTS: Eighty-one patients (71%) had an initial headache of migraine without aura, 13 patients (11.4%) had migraine without aura and coexistent tension-type headache, 11 (9.7%) patients had migraine with and without aura, and 9 patients (7.9%) had episodic tension-type headache. Medications overused by patients included analgesics combined with barbiturates or other nonnarcotic substances in 39.5%, simple analgesics in 38.6%, triptans in 11.4%, and ergotamine in 10.5%. Using the International Headache Society diagnostic criteria, we were able to classify only 28.1% of our patients; the proposed revised criteria for daily and near-daily headaches with medication abuse permitted the classification of 46.4% of patients. CONCLUSION: The minimum dose of medication required to induce chronic headache should be revised because a high proportion of patients are not classifiable using either the International Headache Society diagnostic criteria or the revised criteria recently proposed. A more comprehensive definition for the required minimum dose might be used. Triptan abuse can cause chronic headache and should be included in the International Headache Society classification. PMID- 14756854 TI - Relationship between chronic tension-type headache, cranial hemodynamics, and cerebrospinal pressure: study involving provocation with sumatriptan. AB - OBJECTIVE: To study the relationship between chronic tension-type headache, cranial hemodynamics, and cerebrospinal pressure. BACKGROUND: Cerebrospinal pressure has been found to be above 200 mm in about 50% of patients with chronic tension-type headache. METHODS: Heart rate, blood pressure, common carotid artery diameter and blood flow, and craniovascular resistance and pain at regular intervals before, during, and after head-down tilt-a procedure which increases cerebrospinal pressure, were recorded. After head-down tilt, subcutaneous injections of either placebo or 6 mg of sumatriptan were administered. Chronic tension-type headache intensity before and after withdrawal of 20 mL of cerebrospinal fluid was documented. Cerebrospinal pressure and chronic tension type headache intensity were measured after subcutaneous injection of 6 mg of sumatriptan. RESULTS: Head-down tilt provoked an increase of headache compared with baseline. Common carotid artery blood flow decreased and craniovascular resistance increased after sumatriptan injection, but not after placebo injection. The pain decreased after head-down tilt and placebo injection, but not after sumatriptan injection. Chronic tension-type headache intensity decreased in all 4 patients studied after withdrawal of 20 mL of cerebrospinal fluid. Cerebrospinal pressure increased in 5 patients with chronic tension-type headache after subcutaneous injection of 6 mg of sumatriptan with slight or no increase of pain. CONCLUSION: The results indicated that cerebrospinal pressure or intracranial venous pressure (or both) are related to chronic tension-type headache. PMID- 14756855 TI - Rofecoxib in the prevention of perimenstrual migraine: an open-label pilot trial. AB - OBJECTIVE: To investigate the effectiveness of rofecoxib in the prevention of perimenstrual migraine. BACKGROUND: Nonsteroidal anti-inflammatory drugs have demonstrated benefit in reducing the frequency and intensity of menstrual migraine when administered perimenstrually. Anti-inflammatory drugs, however, may not be well tolerated and can produce gastrointestinal irritation. Rofecoxib, a selective cyclooxygenase-2 inhibitor, has anti-inflammatory and analgesic properties, and a significantly improved gastrointestinal tolerability profile. METHODS: A pilot study was conducted in which patients with a history of menstrually associated migraine and experiencing at least one migraine monthly during the perimenstrual period were enrolled. Patients who completed a baseline diary for the first month were randomized to receive either rofecoxib 25 mg or 50 mg daily for 10 days, starting 5 days before menstrual flow. Headaches experienced during this 10-day period were recorded in the patient's diary. Patients continued rofecoxib for 2 consecutive menstrual cycles. Mean migraine frequency, intensity, and duration as well as patient's level of functioning during the 2 cycles were compared with baseline. RESULTS: Fourteen patients completed baseline and rofecoxib dosing for 2 menstrual cycles. Mean migraine frequency decreased from 5.6 to 2.6 migraines per menstrual cycle (P=.005). Eight (57%) of 14 patients had a > or = 50% reduction in headache frequency. No significant improvement in headache intensity, duration, and functional impairment were observed. Both doses of rofecoxib were well tolerated with no statistical difference in patient response between the doses. CONCLUSION: Rofecoxib at a perimenstrual daily dose of 25 or 50 mg demonstrated a significant reduction in frequency of perimenstrual migraine. A double-blind, placebo controlled trial of rofecoxib in the prevention of perimenstrual migraine is warranted. PMID- 14756856 TI - Cranial magnetic resonance imaging findings in patients with migraine. AB - OBJECTIVE: To investigate the frequency of cranial magnetic resonance imaging abnormalities in patients with migraine and their relationship to type, duration, and frequency of migraine attacks. METHODS: Forty-five patients (43 women, 2 men) with migraine whose ages ranged between 19 and 53 years (mean, 40.91 [SD, 7.69]) were evaluated. Of the 45 patients, 20 had migraine with aura and 25 had migraine without aura, according to the diagnostic criteria of the International Headache Society. RESULTS: In 13 (28.8%) of 45 patients, white matter foci were present on magnetic resonance imaging. Eight of these patients (61.5%) had migraine with aura, and 5 patients (38.4%) had migraine without aura. The presence of white matter foci was significantly higher in the patients with aura (8 [40%] of 20) than in those without aura (5 [20%] of 25). It was found that as the frequency of attacks per month increased, the number of patients with white matter foci also increased. Although the mean duration of migraine was longer in patients with white matter foci (149.5 months [SD, 87.9]) than in those without white matter foci (134.1 months [SD, 88.3]), there was no significant difference (P >.05). CONCLUSION: Although there are no specific magnetic resonance imaging findings peculiar to migraine, detection of white matter foci should be taken into consideration in patients with migraine (especially migraine with aura). Frequency of attacks is an important indicator of existence of white matter foci. PMID- 14756857 TI - Idiopathic intracranial hypertension and postlumbar puncture headache. AB - Idiopathic intracranial hypertension and low cerebrospinal pressure are 2 conditions that are thought to be on opposite ends of the cerebrospinal pressure spectrum. Headache is the prominent component of both conditions. We describe a patient whose evaluation for idiopathic intracranial hypertension resulted in a postlumbar puncture headache. Although not entirely intuitive, we suggest that the 2 conditions can be present in the same patient. PMID- 14756858 TI - Efficacy of topiramate in migraine aura prophylaxis: preliminary results of 12 patients. AB - OBJECTIVE: To evaluate the efficacy of topiramate in the treatment of migraine aura. Antiepileptic drugs may be useful in migraine prevention through such mechanisms as acting directly on the nociceptive system or by modulating the biochemical phenomena of aura. Topiramate acts directly on N-methyl-d-aspartate and amino-3-hydroxy-5-methylisoxazole-4-propionic acid/kainate glutamate receptors and modulates calcium ion channel activity by inhibiting high-voltage activated L-type calcium ion channels. METHODS: Twelve patients with migraine with aura were enrolled in an open-label study and treated with topiramate for 6 months. The dose of topiramate was increased weekly by 25 mg up to a daily dose of 100 mg after 4 weeks. RESULTS: In all 12 patients after 6 months of treatment, topiramate did not statistically influence aura frequency (P=.317) or duration (P=.480) compared with baseline. Mild to moderate side effects were observed, but they did not interfere with treatment. Consistent with previous observations, migraine frequency as well as headache intensity and duration improved statistically significantly. CONCLUSION: Topiramate is not effective in preventing migraine aura. PMID- 14756859 TI - Frequent triptan use: observations on safety issues. AB - OBJECTIVE: To examine the safety of frequent triptan use over extended periods. For a small group of patients with refractory migraine plus chronic daily headache, triptans are effective. METHODS: This retrospective study primarily evaluated the cardiac safety of daily triptan use in 118 patients and, in addition, hematologic tests were assessed. Each patient had utilized a triptan for a minimum of 4 days per week for at least 6 months. Patients with rebound headache had been withdrawn from the triptans. Most patients (97 of 118) averaged 1 tablet daily; most would occasionally go for several days without a triptan. Forty patients had taken a triptan for 6 months to 2 years, 37 patients from 2 to 4 years, and 41 for 4 or more years. RESULTS: Routine hematologic tests were performed periodically on all patients, and no abnormalities were attributable to triptans. Almost all patients had an electrocardiogram, and no abnormal electrocardiograms were felt to be related to triptans. Cardiac echocardiography was performed in 57 patients. The 10 abnormal echocardiograms were not due to triptans. All 20 cardiac stress tests revealed normal findings. Adverse events were minimal; 9 patients described fatigue due to triptans, and 5 had mild chest tightness. CONCLUSION: This long-term study of 118 patients indicates that frequent triptan use may be relatively safe. PMID- 14756860 TI - Cluster headache and the sympathetic nerve. AB - OBJECTIVE: To determine the effect of a sympathetic block at C7 on cluster headache. BACKGROUND: Eleven patients presenting to a pain control unit with cluster headache were included in the study after giving informed consent. METHODS: In all patients, a mixture of 5 mL of 0.5% bupivacaine hydrochloride and 1 cc of methylprednisolone acetate was injected onto the base of the C7 transverse process. RESULTS: The injection was applied during the acute phase of headache in 6 patients and all experienced immediate and complete relief. The other 5 patients received the injection between attacks. Of the 11 patients treated, 8 went into remission by aborting the cluster. In some patients, repeated injections were given before the cluster was aborted. Three patients did not respond to treatment. One patient with chronic paroxysmal hemicrania experienced pain relief of the acute attack after treatment, but the procedure did not abort the subsequent attacks. A surgical sympathectomy removing the stellate ganglion rendered him pain-free for 15 months after which he was lost to follow-up. CONCLUSION: Blocking the sympathetic nerve aborts an acute attack of cluster headache and may play a major role in aborting the cluster. Although only one patient with chronic paroxysmal hemicrania responded to surgical sympathectomy, this procedure may be considered as an alternative if there is poor response to oral medication or a sympathetic block. PMID- 14756861 TI - Migraine and cluster headache: coexistence, laterality, and gender. PMID- 14756863 TI - Fishing for headaches. PMID- 14756862 TI - Acute and persistent nonpositional headache induced by dural trauma and cerebrospinal fluid leak. PMID- 14756864 TI - Industry-sponsored research. PMID- 14756865 TI - Evaluating triptan usage. PMID- 14756866 TI - Tramadol for tension-type headache. PMID- 14756875 TI - Functional gene diversity analysis in BTEX contaminated soils by means of PCR SSCP DNA fingerprinting: comparative diversity assessment against bacterial isolates and PCR-DNA clone libraries. AB - Developments in molecular biology based techniques have led to rapid and reliable tools to characterize microbial community structures and to monitor their dynamics under in situ conditions. However, there has been a distinct lack of emphasis on monitoring the functional diversity in the environment. Genes encoding catechol 2,3-dioxygenases (C23O), as key enzymes of various aerobic aromatic degradation pathways, were used as functional targets to assess the catabolic gene diversity in differentially BTEX contaminated environments by polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP). Site specific PCR-SSCP fingerprints were obtained, showing that gene diversity experienced shifts correlated to temporal changes and levels of contamination. PCR-SSCP enabled the recovery of predominant gene polymorphs, and results closely matched with the information retrieved from random sequencing of PCR-DNA clone libraries. A new method for isolating strains capable of growing on BTEX compounds was developed to diminish preselection or enrichment bias and to assess the function of predominant gene polymorphs. C23O abundance in isolates correlated with the levels of BTEX pollution in the soil samples analysed. Isolates harbouring C23O genes, identical to the gene polymorph predominant in all contaminated sites analysed, showed an unexpected benzene but not toluene mineralizing phenotype whereas isolates harbouring a C23O gene variant differing by a single point mutation and observed in highly polluted sites only, were capable, among some other isolates, to mineralize benzene and toluene, indicating a catabolically determined sharing of carbon sources on-site. The PCR-SSCP technique is thus a powerful tool for assessing the diversity of functional genes and the identification of predominant gene polymorphs in environmental samples as a prerequisite to understand the functioning of microbial communities. PMID- 14756876 TI - Analysis of methanotrophic bacteria in Movile Cave by stable isotope probing. AB - Movile Cave is an unusual groundwater ecosystem that is supported by in situ chemoautotrophic production. The cave atmosphere contains 1-2% methane (CH4), although much higher concentrations are found in gas bubbles that keep microbial mats afloat on the water surface. As previous analyses of stable carbon isotope ratios have suggested that methane oxidation occurs in this environment, we hypothesized that aerobic methane-oxidizing bacteria (methanotrophs) are active in Movile Cave. To identify the active methanotrophs in the water and mat material from Movile Cave, a microcosm was incubated with a 10%13CH4 headspace in a DNA-based stable isotope probing (DNA-SIP) experiment. Using improved centrifugation conditions, a 13C-labelled DNA fraction was collected and used as a template for polymerase chain reaction amplification. Analysis of genes encoding the small-subunit rRNA and key enzymes in the methane oxidation pathway of methanotrophs identified that strains of Methylomonas, Methylococcus and Methylocystis/Methylosinus had assimilated the 13CH4, and that these methanotrophs contain genes encoding both known types of methane monooxygenase (MMO). Sequences of non-methanotrophic bacteria and an alga provided evidence for turnover of CH4 due to possible cross-feeding on 13C-labelled metabolites or biomass. Our results suggest that aerobic methanotrophs actively convert CH4 into complex organic compounds in Movile Cave and thus help to sustain a diverse community of microorganisms in this closed ecosystem. PMID- 14756878 TI - Serial analysis of ribosomal sequence tags (SARST): a high-throughput method for profiling complex microbial communities. AB - Two decades of culture-independent studies have confirmed that microbial communities represent the most complex and concentrated pool of phylogenetic diversity on the planet. There remains a need for innovative molecular tools that can further our knowledge of microbial diversity and its functional implications. We present the method and application of serial analysis of ribosomal sequence tags (SARST) as a novel tool for elucidating complex microbial communities, such as those found in soils and sediments. Serial analysis of ribosomal sequence tags uses a series of enzymatic reactions to amplify and ligate ribosomal sequence tags (RSTs) from bacterial small subunit rRNA gene (SSU rDNA) V1-regions into concatemers that are cloned and sequenced. This approach offers a significant increase in throughput over traditional SSU rDNA clone libraries, as up to 20 RSTs are obtained from each sequencing reaction. To test SARST and measure the bias associated with this approach, RST libraries were prepared from a defined mixture of pure cultures and from duplicate arctic soil DNA samples. The actual RST distribution reflected the theoretical composition of the original defined mixture. Data from duplicate soil libraries (1345 and 1217 RSTs, with 525 and 505 unique RSTs, respectively) indicated that replication provides a strongly correlated RST profile (r(2) = 0.80) and division-level distribution of RSTs (r(2) = 0.99). Using sequence data from abundant soil RSTs, we designed specific primers that successfully amplified a larger portion of the SSU rDNA for further phylogenetic analysis. These results suggest that SARST is a powerful approach for reproducible high-throughput profiling of microbial diversity amenable to medical, industrial or environmental microbiology applications. PMID- 14756877 TI - Host specificity in marine sponge-associated bacteria, and potential implications for marine microbial diversity. AB - Biodiversity is fundamental to both eukaryote and prokaryote ecology, yet investigations of diversity often differ markedly between the two disciplines. Host specificity - the association of organisms with only a few (specialism) or many (generalism) host species - is recognized within eukaryote ecology as a key determinant of diversity. In contrast, its implications for microbial diversity have received relatively little attention. Here we explore the relationship between microbial diversity and host specificity using marine sponge-bacteria associations. We used a replicated, hierarchical sampling design and both 16S rDNA- and rpoB-based denaturing gradient gel electrophoresis (DGGE) to examine whether three co-occurring sponges from temperate Australia -Cymbastela concentrica, Callyspongia sp. and Stylinos sp. - contained unique, specialized communities of microbes. Microbial communities varied little within each species of sponge, but variability among species was substantial. Over five seasons, the microbial community in C. concentrica differed significantly from other sponges, which were more similar to seawater. Overall, three types of sponge-associated bacteria were identified via 16S rDNA sequencing of excised DGGE bands: 'specialists'- found on only one host species, 'sponge associates'- found on multiple hosts but not in seawater, and 'generalists' from multiple hosts and seawater. Analogous to other high diversity systems, the degree of specificity of prokaryotes to host eukaryotes could have a potentially significant effect on estimates of marine microbial diversity. PMID- 14756879 TI - Genomic polymorphism in symbiotic populations of Photobacterium leiognathi. AB - Photobacterium leiognathi forms a bioluminescent symbiosis with leiognathid fishes, colonizing the internal light organ of the fish and providing its host with light used in bioluminescence displays. Strains symbiotic with different species of the fish exhibit substantial phenotypic differences in symbiosis and in culture, including differences in 2-D PAGE protein patterns and profiles of indigenous plasmids. To determine if such differences might reflect a genetically based symbiont-strain/host-species specificity, we profiled the genomes of P. leiognathi strains from leiognathid fishes using PFGE. Individual strains from 10 species of leiognathid fishes exhibited substantial genomic polymorphism, with no obvious similarity among strains; these strains were nonetheless identified as P. leiognathi by 16S rDNA sequence analysis. Profiling of multiple strains from individual host specimens revealed an oligoclonal structure to the symbiont populations; typically one or two genomotypes dominated each population. However, analysis of multiple strains from multiple specimens of the same host species, to determine if the same strain types consistently colonize a host species, demonstrated substantial heterogeneity, with the same genomotype only rarely observed among the symbiont populations of different specimens of the same host species. Colonization of the leiognathid light organ to initiate the symbiosis therefore is likely to be oliogoclonal, and specificity of the P. leiognathi/leiognathid fish symbiosis apparently is maintained at the bacterial species level rather than at the level of individual, genomotypically defined strain types. PMID- 14756880 TI - Physiological and molecular genetic evaluation of the dechlorination agent, pyridine-2,6-bis(monothiocarboxylic acid) (PDTC) as a secondary siderophore of Pseudomonas. AB - The bacterial metabolite and transition metal chelator pyridine-2,6 dithiocarboxylic acid (PDTC), promotes a novel and effective means of dechlorination of the toxic and carcinogenic pollutant, carbon tetrachloride. Pyridine-2,6-dithiocarboxylic acid has been presumed to act as a siderophore in the Pseudomonas strains known to produce it. To explore further the physiological function of PDTC production, we have examined its regulation, the phenotype of PDTC-negative (pdt) mutants, and envelope proteins associated with PDTC in P. putida strain DSM 3601. Aspects of the regulation of PDTC production and outer membrane protein composition were consistent with siderophore function. Pyridine 2,6-dithiocarboxylic acid production was coordinated with production of the well characterized siderophore pyoverdine; exogenously added pyoverdine led to decreased PDTC production, and added PDTC led to decreased pyoverdine production. Positive regulation of a chromosomal pdtI-xylE transcriptional fusion, and of a 66 kDa outer membrane protein (IROMP), was seen in response to exogenous PDTC. Tests with transition metal chelators indicated that PDTC could provide a benefit under conditions of metal limitation; the loss of PDTC biosynthetic capacity caused by a pdtI transposon insertion resulted in increased sensitivity to 1,10 phenanthroline, a chelator that has high affinity for a range of divalent transition metals (e.g. Fe(2+), Cu(2+), Zn(2+)). Exogenously added PDTC could also suppress a phenotype of pyoverdine-negative (Pvd-) mutants, that of sensitivity to EDDHA, a chelator with higher affinity and specificity for Fe(3+). Measurement of 59Fe incorporation showed uptake from 59Fe:PDTC by DSM 3601 grown in low-iron medium, but not by cells grown in high iron medium, or by the pdtI mutant, which did not show expression of the 66 kDa envelope protein. These data verified a siderophore function for PDTC, and have implicated it in the uptake of transition metals in addition to iron. PMID- 14756881 TI - Design of 16S rRNA-targeted oligonucleotide probes for detecting cultured and uncultured archaeal lineages in high-temperature environments. AB - In order to facilitate the evaluation of archaeal community diversity and distribution in high-temperature environments, 14 16S rRNA oligonucleotide probes were designed. Adequate hybridization and wash conditions of the probes encompassing most known hyperthermophilic Archaea, members of the orders Thermococcales, Desulfurococcales and Sulfolobales, of the families Methanocaldococcaceae, Pyrodictiaceae and Thermoproteaceae, of the genera Archaeoglobus, Methanopyrus and Ignicoccus, and of the as yet uncultured lineages Korarchaeota, Crenarchaeota marine group I, deep-sea hydrothermal vent euryarchaeotic group 2 (DHVE 2), and deep-sea hydrothermal vent euryarchaeotic group 8 (DHVE 8) were determined by dot-blot hybridization from target and non target reference organisms and environmental clones. The oligonucleotide probes were also used to evaluate the archaeal community composition in nine deep-sea hydrothermal vent samples. All probes, except those targeting members of Sulfolobales, Thermoproteaceae, Pyrodictiaceae and Korarchaeota, gave positive hybridization signals when hybridized against 16S rDNA amplification products obtained from hydrothermal DNA extracts. The results confirmed the widespread occurrence of Thermococcales, Desulfurococcales, Methanocaldococcaceae and Archaeoglobus in deep-sea hydrothermal vents, and extended the known ecological habitats of uncultured lineages. Despite their wide coverage, the probes were unable to resolve the archaeal communities associated with hydrothermally influenced sediments, suggesting that these samples may contain novel lineages. This suite of oligonucleotide probes may represent an efficient tool for rapid qualitative and quantitative characterization of archaeal communities. Their application would help to provide new insights in the future into the composition, distribution and abundance of Archaea in high-temperature environments. PMID- 14756882 TI - A novel approach to investigate biofilm accumulation and bacterial transport in porous matrices. AB - Knowledge of bacterial transport through, and biofilm growth in, porous media is vitally important in numerous natural and engineered environments. Despite this, porous media systems are generally oversimplified and the local complexity of cell transport, biofilm formation and the effect of biofilm accumulation on flow patterns is lost. In this study, cells of the sulphate-reducing bacterium, Desulfovibrio sp. EX265, accumulated primarily on the leading faces of obstructions and developed into biofilm, which grew to narrow and block pore throats (at a rate of 12 micro m h(-1) in one instance). This pore blocking corresponded to a decrease in permeability from 9.9 to 4.9 Darcy. Biofilm processes were observed in detail and quantitative data were used to describe the rate of biofilm accumulation temporally and spatially. Accumulation in the inlet zone of the micromodel was 10% higher than in the outlet zone and a mean biofilm height of 28.4 micro m was measured in a micromodel with an average pore height of 34.9 microm. Backflow (flow reversal) of fluid was implemented on micromodels blocked with biofilm growth. Although biofilm surface area cover did immediately decrease (approximately 5%), the biofilm quickly re-established and permeability was not significantly affected (9.4 Darcy). These results demonstrate that the glass micromodel used here is an effective tool for in situ analysis and quantification of bacteria in porous media. PMID- 14756885 TI - Dermatologic therapy of chronic genital disease. AB - The adaptation of dermatology for the genital area requires several modifications of standard therapy. This unique area produces issues of both psychological sensitivity and unique environmental factors of constant moisture, warmth, and friction. These issues become far more important when symptoms are chronic. Recognition of normal variants, the multifactorial nature of many genital symptoms, and the avoidance of creating secondary iatrogenic disease are all important. PMID- 14756886 TI - Lichen simplex chronicus (atopic/neurodermatitis) of the anogenital region. AB - Lichen simplex chronicus (LSC) of the anogenital area is an eczematous disease characterized by unremitting itching and scratching. In most instances, it arises in individuals who are genetically atopic, and as such, LSC can be viewed as a localized variant of atopic/neurodermatitis. Common triggers for the development of the disease include psychological distress, and local environmental problems such as heat, sweating, and excess dryness. Lichen simplex chronicus may also develop as a superimposed condition in the presence of other anogenital diseases such as candidiasis, psoriasis, lichen sclerosus, tinea cruris, and neoplasia. Lichen simplex chronicus frequently persists as an itch-scratch cycle, even when environmental triggers are removed and the underlying disease is treated. For this reason, successful therapy requires attention not only to trigger factors, but also to repair of the damaged barrier layer, reduction in inflammation, and breakup of the itch-scratch cycle. PMID- 14756887 TI - Contact dermatitis of the vulva. AB - Vulvar diseases rarely stand alone. They are often caused or worsened by primary irritant or allergic contact dermatitis, and this should be considered when evaluating any vulvar complaint. All irritants should be avoided in all women, and those with vulvar dermatoses should be patch tested to help define or rule out allergens. PMID- 14756888 TI - Lichen sclerosus: a review and practical approach. AB - Lichen sclerosus (LS) is a chronic dermatitis predominantly found in the anogenital area. It can be found in patients of any age group, sex, or race, but is most commonly present in Caucasian peri- or postmenopausal women. Although the etiology of LS remains uncertain, an autoimmune process is believed to underlie this condition. With many cases going unreported, its incidence is still unknown. There is no cure for LS, but treatment offers control of the condition. They are three reasons for treating LS: relief of symptoms and discomfort; prevention of any or further anatomical changes; and a theoretical prevention of malignant transformation. Although many treatments have been suggested to treat LS over the years, only potent or ultra-potent corticosteroids remain as the treatment of choice. After initial therapy, some patients might only use corticosteroids as needed, while others may require a twice-weekly maintenance therapy. There is no place for surgery in uncomplicated LS. Surgery should be limited exclusively to patients with malignancy and to correct scarring secondary to the disease. Lichen sclerosus is associated with a 4-6% risk of squamous cell carcinoma, making long term follow-up essential in these patients. PMID- 14756889 TI - Diagnosis and therapy of anogenital lichen planus. AB - Genital lichen planus is a fairly common disease of genital skin that exhibits a widely variable morphology. Lichen planus occurring on the mucosal surfaces is chronic, and most often erosive and painful, frequently producing debilitating scarring. Mild, non-erosive disease is controllable with topical corticosteroids. Although erosive disease is much more difficult to manage, the use of topical corticosteroids, sometimes in combination with other topical or systemic medications, maximizes comfort and preserves function, especially with careful attention to local care and secondary infection. PMID- 14756890 TI - Desquamative inflammatory vaginitis. AB - Desquamative inflammatory vaginitis (DIV) is not a diagnosis in itself, and may be the presentation of a range of blistering disorders including pemphigus vulgaris, lichen planus and mucous membrane pemphigoid. The existence of an idiopathic subset of DIV remains controversial and is discussed in the present article. Desquamative inflammatory vaginitis is a rare but disabling condition. It presents in women of any age with a history of discomfort, irritation and painful sexual intercourse. Patients may also report an increased vaginal discharge. Examination of the vulva is normal, but erythematous regions on the vaginal walls are evident with increased vaginal secretion. This secretion is high in polymorphonuclear leukocytes, with an increased number of immature squamous epithelial cells. Repeated cultures are negative for bacteria, viruses and yeast. This is a sterile inflammatory vaginitis that can be difficult to treat, but successful therapy has been reported with topical steroids and clindamycin. PMID- 14756891 TI - Hidradenitis suppurativa: a review. AB - Hidradenitis suppurativa is a recurrent disease involving apocrine-bearing skin with a predilection for intertriginous areas, including genital skin. It has a highly variable clinical course. Mild cases may present as recurrent isolated nodules, while severe instances of the disease with chronic inflammation may lead to scarring, functional impairment, and rarely, squamous cell carcinoma. While genetic factors, patient characteristics, hormones and infection play a role in disease expression, a comprehensive understanding of the pathogenesis remains to be elicited. Additionally, effective treatment is largely unknown. While the mainstay of therapy had been surgery, and topical or systemic antimicrobial agents, other therapeutic modalities such as retinoids, hormonal therapy and immunosuppresive medications may also hold some promise. PMID- 14756892 TI - Vulvar ulcers and erosions--a dermatologist's viewpoint. AB - Vulvar ulceration can occur as the primary or secondary event in a large variety of conditions. These include infections, autoimmune and/or inflammatory diseases and dermatoses, neoplasias, and conditions with an unknown etiology. A thorough medical history and careful patient examination remain the linchpin of management. Specific microbiological, histological, immunological, and other investigations are often necessary to establish or confirm a diagnosis, but the relevant importance of these will vary in the individual patient. The specific management of each patient will also vary accordingly. Other important factors which will influence the frequency with which any physician will make one of the specific diagnoses include practice location, referral mechanisms, and population demographics. This chapter is written with the generalist dermatologist in mind. Whilst dermatoses are given prime attention, important infections and other conditions are also dealt with. The ultimate aim of the present paper is to provide the generalist dermatologist with a useful tool for the diagnosis and management of a patient that present's with a vulvar ulcer(s). PMID- 14756893 TI - Therapy of external anogenital warts and molluscum contagiosum: a literature review. AB - Anogenital warts and mollusca contagiosum are virally induced, benign skin tumors for which there is no single preferable therapy. Treatments include physical and chemical destruction, surgical removal, and biological response modifiers to enhance the natural immune response. The choice of therapy is an art, and depends upon patient preference, finances, number of lesions, and lesional morphology. However, the therapy of these lesions can sometimes be very painful and expensive, and therapy should not be worse than the disease. PMID- 14756894 TI - The diagnosis and treatment of infectious vaginitis. AB - Inflammation of the vagina as a result of infectious agents is very common, both as an overgrowth of normal or common colonizers, or as a frank infection. The most common causes of infectious vaginitis are yeast, bacteria, protozoa, viruses, and parasites. Infections of the vagina produce an increase in vaginal secretion, vulvar symptoms of itching or irritation from contact with irritating vaginal fluid, and sometimes odor. A careful microscopic examination of vaginal secretions generally yields the correct diagnosis, but atypical or recalcitrant disease deserves a confirmatory culture, as noninfectious inflammatory processes can produce similar symptoms. PMID- 14756895 TI - Vulvar fissures: causes and therapy. AB - Vulvar fissures occur in two main patterns: at the posterior fourchette, and within skin folds and creases. The cause of posterior fourchette splitting is not known, and the treatment is a perineoplasty. Skin-fold fissures occur in response to several inflammatory dermatoses or infections, and therapy consists of elimination of any underlying infection and the (sometimes prolonged) use of a topical corticosteroid ointment. PMID- 14756896 TI - Therapy of pediatric genital diseases. AB - Infants and children have special issues with regard to genital disease. Infants are incontinent, and have an increase in local irritation and infection risk. In addition, the adult sex hormones which enhance the health of genital skin are deficient. Also, the choice of therapy must be modified to take into account the more fragile nature of prepubertal skin, the tolerance of children to painful treatments, and the lack of experience of some medications in children. PMID- 14756897 TI - An approach to the treatment of anogenital pruritus. AB - The anogenital area is a common location for pruritic complaints. Specific terms for chronic itch in this location have included pruritus vulvae, pruritus ani, lichen simplex chronicus, and neurodermatitis. A male counterpart to pruritus vulvae, pruritus scroti, is less common. Acute anogenital pruritus is usually caused by infections or contact dermatitis. In chronic pruritus, inflammatory dermatoses and malignancies must be ruled out. In idiopathic anogenital pruritus or neurodermatitis, the skin findings should be limited to lichenification and excoriations. Skin findings may be entirely absent. When treating anogenital pruritus, topic irritants and potential sensitizers must be eliminated. Cleansing and toilet habits must be addressed. A short course of a high-potency topical steroid should bring moderate to complete relief. Sedating antihistamines may limit nighttime symptoms. In some patients, psychotropic agents are required to achieve adequate sedation. Antidepressants may be required in patients refractory to treatment or with underlying psychiatric disorders. PMID- 14756898 TI - Management of vulvar pain. AB - Vulvodynia is a frequently used medical term that literally means "vulvar pain". Therefore, vulvodynia is a symptom, not a disease. The term itself indicates a variety of unpleasant chronic vulvar sensations, including burning, rawness, soreness, irritation, sensitivity, and formication. This may or may not include dyspareunia. Primary vulvodynia occurs when these sensory disturbances occur in the absence of observable dermatologic disease or vulvovaginal infection. There are several causes for this, including neuropathy, referred pain, and pelvic floor muscle dysfunction. For the purist, it is the patient in whom there is no observable reason for vulvar pain who represents the true case of vulvodynia. However, vulvodynia can also occur secondarily as a symptom of vulvar skin disease. Restricting the present paper to patients without objective signs leaves out all the important conditions which come into the differential diagnosis of vulvar pain which should be ruled out first. The first step in managing vulvodynia is making an accurate diagnosis of its cause. The present review summarizes the diagnosis and management of the chronic dermatologic diseases which may cause primary and secondary vulvodynia. The etiology of primary vulvodynia is much more poorly understood than secondary vulvodynia, and treatment of some aspects remains controversial. PMID- 14756899 TI - The effects of chemotherapeutics on cellular metabolism and consequent immune recognition. AB - Awidely held view is that oncolytic agents induce death of tumor cells directly. In this report we review and discuss the apoptosis-inducing effects of chemotherapeutics, the effects of chemotherapeutics on metabolic function, and the consequent effects of metabolic function on immune recognition. Finally, we propose that effective chemotherapeutic and/or apoptosis-inducing agents, at concentrations that can be achieved physiologically, do not kill tumor cells directly. Rather, we suggest that effective oncolytic agents sensitize immunologically altered tumor cells to immune recognition and immune-directed cell death. PMID- 14756900 TI - The efficacy and safety of Chinese herbal medicines. PMID- 14756901 TI - Is the synthesis of rumen bacterial protein limited by the availability of pre formed amino acids and/or peptides? PMID- 14756902 TI - Is zinc deficiency a risk factor for atherosclerosis? AB - The development of atherosclerosis is influenced by genetic, lifestyle and nutritional risk factors. Zn and metallothionein deficiency can enhance oxidative stress-related signalling processes in endothelial cells, and since changes in available plasma Zn may affect the Zn status of the endothelium, Zn deficiency could be a risk factor for IHD. Although the association of Zn with many proteins is essential for their function, three key signalling processes are highlighted as being principal targets for the effect of Zn deficiency: the activation of NF kappaB, the activation of caspase enzymes and the signalling of NO. The need to develop a reliable indicator of Zn status is critical to any epidemiological approach for studying the relationship between Zn status and disease incidence. Studies using appropriate animal models and investigating how the plasma Zn pool influences endothelial intracellular labile Zn would be helpful in appreciating the importance of Zn deficiency in atherogenesis. PMID- 14756903 TI - The relationships between potassium intakes, transmural potential difference of the rumen epithelium and magnesium absorption in wethers. AB - In vitro studies with isolated sheep rumen epithelium have shown that an increase in the lumen K concentration induces an increase in the transmural potential difference across the rumen epithelium (serosal side: positive), which is associated with a decrease in Mg transport. However, at lumen K concentrations >80 mmol/l, Mg transport across the epithelium became independent of the lumen K concentration. The present study was carried out to determine whether this observation also occurs in vivo. Four ruminally fistulated wethers were fed four rations supplemented with KHCO3 (15.7, 37.6, 59.4 or 77.4 g K/kg DM) in a 4 x 4 Latin square design. Increased K intakes significantly increased the rumen K concentration. For all data combined, Mg absorption expressed as % intake was negatively correlated with the rumen K concentration. However, apparent Mg absorption either expressed in absolute terms (g/d) or as % intake was not significantly affected when the dietary K concentration was increased from 59.4 to 77.4 g/kg DM. Rumen K concentration was inversely correlated with the transmural potential difference (blood side: positive) (Pearson's r -0.709; R(2)adj 0.468, P=0.002, n 16). It is concluded that in wethers apparent Mg absorption becomes independent of the dietary K concentration when the K concentration is >60 g/kg DM or equivalent to a postprandial rumen K concentration of about 125 mmol/l. PMID- 14756904 TI - Lipid atherogenic risk markers can be more favourably influenced by the cis 9,trans-11-octadecadienoate isomer than a conjugated linoleic acid mixture or fish oil in hamsters. AB - The aim of our present study was to compare the efficiency of conjugated linoleic acids (CLA) and fish oil in modulating atherogenic risk markers. Adult male hamsters were given a cholesterol-rich diet (0.6 g/kg) for 8 weeks; the diet was supplemented with 5 g cis-9,trans-11-CLA isomer/kg, 12 g CLA mixture (CLA mix)/kg, 12 g fish oil/kg or 12 g fish oil+12 g CLA-mix/kg. The plasma cholesterol status was improved only with the cis-9,trans-11-CLA (HDL-cholesterol and HDL-cholesterol:LDL-cholesterol ratio, P<0.05), but was of borderline significance for CLA-mix (HDL-cholesterol:LDL-cholesterol ratio, P=0.06), with an increase (33-40 %) in the liver lipoprotein receptors (scavenger receptor-type I and LDL ApoB/E receptor) and HDL-binding protein 2 (P<0.05). A 100 % pigment gallstones incidence and a slight insulin resistance (homeostatic model assessment index) were observed in the CLA-mix-fed hamsters (P=-0.031). In comparison, fish-oil feeding alone improved merely the scavenger receptor-type I and HDL-binding protein 2 liver status and faeces sterol output. For most of our present observations, the concomitant intake of fish oil and CLA-mix gave dominant effects that were exclusive and specific to one or the other oil. In conclusion, part of the beneficial effects of CLA in the present study can be ascribed to the cis-9,trans-11-isomer, and these did not generally overlap with those of fish oil. In addition, the CLA-mix effects are clearly affected by the marine (n-3) fatty acids. PMID- 14756905 TI - Disaccharidase activity in the intestinal tract of Wistar-Furth, diabetes resistant and diabetes-prone BioBreeding rats. AB - Diabetes-prone BioBreeding (BBdp) rats often present an enteropathy that may precede the onset of autoimmune insulitis. The aim of the present study was to assess the influence of sex, the time course, the strain specificity, the distribution along the intestinal tract and the effect of diet for the changes in the activity of gut invertase, maltase and lactase found in BBdp rats, as compared with both Wistar-Furth (WF) and diabetes-resistant BioBreeding (BBc) rats. These hydrolases were measured, therefore, at day 10, 30, 45, 70, 95 and 120 in three intestinal segments of WF, BBc and BBdp rats fed, after weaning, either a protective hydrolysed casein diet, which decreases the incidence of diabetes in the BBdp rats, or one of two diabetogenic diets (National Toxicology Program; NTP or wheat-gluten-based; WG) [corrected]. Except for a somewhat lower lactase activity in the BBdp rats, no obvious difference in hydrolyase activity between the three strains of rats was observed at day 10. Between days 30 and 120, however, the activity of the hydrolases, especially that of invertase and lactase, was lower in the BBdp rats than in either the WF or BBc rats, at least when considering the animals fed either the NTP or WG diet. These findings support the view that BBdp rats exposed to a diabetogenic diet develop an enteropathy well before the onset of autoimmune insulitis, in a manner somehow comparable with the situation found in some type 1 diabetic patients, in whom coeliac disease may be diagnosed before diabetes onset. PMID- 14756907 TI - Effects of structured medium- and long-chain triacylglycerols in diets with various levels of fat on body fat accumulation in rats. AB - The effects of structured medium- and long-chain triacylglycerols (MLCT) in diets containing 50-200 g fat/kg on body fat accumulation were compared with those of long-chain triacylglycerols (LCT) in rats. In rats fed ad libitum, weights of intra-abdominal adipose tissues and carcass fat contents were significantly smaller (P<0.05) in rats fed the 150-200 g MLCT/kg diet than in rats fed 150-200 g LCT/kg diet. Serum and liver triacylglycerol contents were significantly greater (P<0.05) in rats fed 200 g MLCT/kg diet, as were hepatic capacities of citrate synthase and cytochrome oxidase (P<0.05). The effects of MLCT on body fat were also examined in adult rats fed a limited amount of food (approximately 50 % of ad libitum intake). Reduction of body fat deposition during the food restriction was the same between in LCT and MLCT groups. These results suggest that accumulation of body fat was less efficient during long-term feeding of MLCT than LCT in rats fed high-fat diets ad libitum. The effect of MLCT on body fat might be influenced by the dietary fat content or by energy sufficiency. PMID- 14756906 TI - Consumption of brown onions (Allium cepa var. cavalier and var. destiny) moderately modulates blood lipids, haematological and haemostatic variables in healthy pigs. AB - Although garlic and onions have long been associated with putative cardiovascular health benefits, the effects of different commercially available onions and level of intake have not been studied. Therefore, the aim of the present study was to evaluate the potential health benefits of raw onions using the pig as a biomedical model. Twenty-five female (Large White x Landrace) pigs were used in a (2 x 2)+1 factorial experiment. Pigs were fed a standard grower diet supplemented with 100 g tallow/kg with the addition of Allium cepa var. cavalier or var. destiny at 0, 10 or 25 g/MJ digestible energy for 6 weeks. Overall, the consumption of onions resulted in significant reductions in plasma triacylglycerol; however, the reductions were most pronounced in pigs fed destiny onions (-26 %, P=0.042). Total plasma cholesterol and LDL:HDL ratios were not significantly different. Onion supplementation, regardless of the variety, resulted in dose-dependent reductions in erythrocyte counts and Hb levels, while the white blood cell concentrations, particularly lymphocytes, were increased in pigs that consumed onions. Furthermore, indices of blood clotting were largely unaffected by onion consumption. In conclusion, dietary supplementation with raw brown onions has moderate lipid-modulating and immunostimulatory properties. However, daily onion intake >25 g/MJ digestible energy could be detrimental to erythrocyte numbers. PMID- 14756908 TI - Dietary-induced changes in the fatty acid profile of rat pancreatic membranes are associated with modifications in acinar cell function and signalling. AB - The effects of dietary lipids on the fatty acid composition of rat pancreatic membranes and acinar cell function were investigated. Weaning rats were fed for 8 weeks on one of two diets which contained 100 g virgin olive oil (OO) or sunflower-seed oil (SO)/kg. Pancreatic plasma membranes were isolated and fatty acids determined. Amylase secretion and cytosolic concentrations of Ca(2+) and Mg(2+) were measured in pancreatic acini. Membrane fatty acids were profoundly affected by the diets; the rats fed OO had higher levels of 18 : 1n-9 (42.86 (sem 1.99) %) and total MUFA compared with the animals fed SO (25.37 (sem 1.11) %). Reciprocally, the SO diet resulted in greater levels of total and n-6 PUFA than the OO diet. The most striking effect was observed for 18 : 2n-6 (SO 17.88 (sem 1.32) %; OO 4.45 (sem 0.60) %), although the levels of 20 : 4n-6 were also different. The proportion of total saturated fatty acids was similar in both groups, and there was only a slight, not significant (P=0.098), effect on the unsaturation index. Compared with the OO group, acinar cells from the rats fed SO secreted more amylase at rest but less in response to cholecystokinin octapeptide, and this was paralleled by reduced Ca(2+) responses to the secretagogue. The results confirm that rat pancreatic cell membranes are strongly influenced by the type of dietary fat consumed and this is accompanied by a modulation of the secretory activity of pancreatic acinar cells that involves, at least in part, Ca(2+) signalling. PMID- 14756909 TI - The use of uniaxial accelerometry for the assessment of physical-activity-related energy expenditure: a validation study against whole-body indirect calorimetry. AB - Assessing the total energy expenditure (TEE) and the levels of physical activity in free-living conditions with non-invasive techniques remains a challenge. The purpose of the present study was to investigate the accuracy of a new uniaxial accelerometer for assessing TEE and physical-activity-related energy expenditure (PAEE) over a 24 h period in a respiratory chamber, and to establish activity levels based on the accelerometry ranges corresponding to the operationally defined metabolic equivalent (MET) categories. In study 1, measurement of the 24 h energy expenditure of seventy-nine Japanese subjects (40 (SD 12) years old) was performed in a large respiratory chamber. During the measurements, the subjects wore a uniaxial accelerometer (Lifecorder; Suzuken Co. Ltd, Nagoya, Japan) on their belt. Two moderate walking exercises of 30 min each were performed on a horizontal treadmill. In study 2, ten male subjects walked at six different speeds and ran at three different speeds on a treadmill for 4 min, with the same accelerometer. O2 consumption was measured during the last minute of each stage and was expressed in MET. The measured TEE was 8447 (SD 1337) kJ/d. The accelerometer significantly underestimated TEE and PAEE (91.9 (SD 5.4) and 92.7 (SD 17.8) % chamber value respectively); however, there was a significant correlation between the two values (r 0.928 and 0.564 respectively; P<0.001). There was a strong correlation between the activity levels and the measured MET while walking (r(2) 0.93; P<0.001). Although TEE and PAEE were systematically underestimated during the 24 h period, the accelerometer assessed energy expenditure well during both the exercise period and the non-structured activities. Individual calibration factors may help to improve the accuracy of TEE estimation, but the average calibration factor for the group is probably sufficient for epidemiological research. This method is also important for assessing the diurnal profile of physical activity. PMID- 14756910 TI - The acute effects of olive oil v. cream on postprandial thermogenesis and substrate oxidation in postmenopausal women. AB - The influence of the source of dietary fat on postprandial thermogenesis and substrate oxidation rates, was examined in twelve postmenopausal women aged 57-73 years, with BMI 21.9-38.3 kg/m(2). A single blind, randomised, paired comparison of two high-fat, isoenergetic, mixed test meals was conducted. The major source of fat was either cream (CREAM) or extra virgin olive oil (EVOO). RMR, diet induced thermogenesis (DIT) and substrate oxidation rates over 5 h were measured by indirect calorimetry. There were no differences in body weight, RMR, fasting carbohydrate or fat oxidation rates between the two occasions. DIT (EVOO 97 (SD 46) v. CREAM 76 (SD 69) kJ/5 h and EVOO 5.2 (SD 2.5) v. CREAM 4.1 (SD 3.7)% energy) did not differ between the two test meals. The postprandial increase in carbohydrate oxidation rates, relative to their respective fasting values (DeltaCOX), was significantly lower following the EVOO meal (EVOO 10.6 (SD 8.3) v. CREAM 17.5 (SD 10) g/5 h; paired t test, P=0.023), while postprandial fat oxidation rates (DeltaFOX) were significantly higher (EVOO 0.0 (SD 4.4) v. CREAM 3.6 (sd 4.0) g/5 h; P=0.028). In the eight obese subjects, however, DIT was significantly higher following the EVOO meal (EVOO 5.1 (SD 2.0) v. CREAM 2.5 (sd 2.9) %; P=0.01). This was accompanied by a significantly lower DeltaCOX (EVOO 10.9 (SD 9.9) v. CREAM 17.3 (SD 10.5) g/5 h; P=0.03) and significantly higher DeltaFOX (EVOO 0.11 (SD 4.4) v. CREAM -4.1 (SD 4.5) g/5 h, P=0.034). The present study showed that olive oil significantly promoted postprandial fat oxidation and stimulated DIT in abdominally obese postmenopausal women. PMID- 14756911 TI - Use of stable isotopes to measure de novo synthesis and turnover of amino acid-C and -N in mixed micro-organisms from the sheep rumen in vitro. AB - Protein synthesis and turnover in ruminal micro-organisms were assessed by stable isotope methods in order to follow independently the fate of amino acid (AA)-C and -N in different AA. Rumen fluid taken from sheep receiving a grass hay concentrate diet were strained and incubated in vitro with starch-cellobiose xylose in the presence of NH3 and 5 g algal protein hydrolysate (APH)/l, in incubations where the labels were (15)NH3, [(15)N]APH or [(13)C]APH. Total (15)N incorporation was calculated from separate incubations with (15)NH3 and [(15)N]APH, and net N synthesis from the increase in AA in protein-bound material. The large difference between total and net AA synthesis indicated that substantial turnover of microbial protein occurred, averaging 3.5 %/h. Soluble AA N was incorporated on average more extensively than soluble AA-C (70 v. 50 % respectively, P=0.001); however, incorporation of individual AA varied. Ninety percent of phenylalanine-C was derived from the C-skeleton of soluble AA, whereas the incorporation of phenylalanine-N was 72 %. In contrast, only 15 % aspartate C+asparagine-C was incorporated, while 45 % aspartate-N+asparagine-N was incorporated. Deconvolution analysis of mass spectra indicated substantial exchange of carboxyl groups in several AA before incorporation and a condensation of unidentified C2 and C4 intermediates during isoleucine metabolism. The present results demonstrate that differential labelling with stable isotopes is a way in which fluxes of AA synthesis and degradation, their biosynthetic routes, and separate fates of AA-C and -N can be determined in a mixed microbial population. PMID- 14756912 TI - Ganoderma lucidum ("Lingzhi"), a Chinese medicinal mushroom: biomarker responses in a controlled human supplementation study. AB - Lingzhi (Ganoderma lucidum) is a woody mushroom highly regarded in traditional medicine and is widely consumed in the belief that it promotes health and longevity, lowers the risk of cancer and heart disease and boosts the immune system. However, objective scientific validation of the putative health benefits of Lingzhi in human subjects is lacking, and issues of possible toxicity must be addressed. The present double-blinded, placebo-controlled, cross-over intervention study investigated the effects of 4 weeks Lingzhi supplementation on a range of biomarkers for antioxidant status, CHD risk, DNA damage, immune status, and inflammation, as well as markers of liver and renal toxicity. It was performed as a follow-up to a study that showed that antioxidant power in plasma increased after Lingzhi ingestion, and that 10 d supplementation was associated with a trend towards an improved CHD biomarker profile. In the present study, fasting blood and urine from healthy, consenting adults (n 18; aged 22-52 years) was collected before and after 4 weeks supplementation with a commercially available encapsulated Lingzhi preparation (1.44 g Lingzhi/d; equivalent to 13.2 g fresh mushroom/d) or placebo. No significant change in any of the variables was found, although a slight trend toward lower lipids was again seen, and antioxidant capacity in urine increased. The results showed no evidence of liver, renal or DNA toxicity with Lingzhi intake, and this is reassuring. The present study of the effects in healthy, well-nourished subjects provides useful, new scientific data that will support controlled intervention trials using at-risk subjects in order to assess the therapeutic effect of Lingzhi in the promotion of healthy ageing. PMID- 14756913 TI - Supplementation of grazing dairy cows with rumen-protected tuna oil enriches milk fat with n-3 fatty acids without affecting milk production or sensory characteristics. AB - The present study was conducted to determine the pattern of incorporation of dietary EPA and docosahexaenoic acid (DHA) into milk, and to evaluate consequent changes in milk fat composition and sensory characteristics. Fourteen multiparous cows in early lactation were divided into two groups and were offered supplements for 10 d. While individual stalls after each morning milking, one group was offered a mixture of rumen-protected tuna oil (RPTO)-soyabean supplement (2 kg; 30:70, w/w; +RPTO) and the second group was offered the basal ration without RPTO (-RPTO). Both groups grazed together on a spring pasture after supplementation. Feeding supplemental RPTO increased the concentrations of EPA and DHA in milk fat from undetectable levels in -RPTO cows to 6.9 and 10.1 g/kg milk fat respectively. Total n-3 PUFA concentration in milk fat was increased three- to fourfold by tuna-oil supplementation (8.4 to 32.0 g/kg milk fat). There were no significant effects on milk production (35.4 v. 33.9 l/d), milk protein (28.2 v. 30.1 g/kg) or milk fat (36.2 v. 40.4 g/kg for -RPTO and +RPTO respectively). The concentration of total saturated fatty acids in milk fat was significantly reduced (568 v. 520 g/kg total fatty acids) and there was a 17 % reduction in the atherosclerotic index of milk after tuna-oil supplementation. Untrained consumer panellists (n 61) rated milk from both groups of cows similarly for taste and smell. We conclude that it is possible to enrich milk with n-3 PUFA without deleterious effects on yield, milk composition or sensory characteristics. PMID- 14756914 TI - Mental and psychomotor development in Indonesian infants of mothers supplemented with vitamin A in addition to iron during pregnancy. AB - Maternal nutrition is important for fetal development, but its impact on the functional outcome of infants is still unclear. The present study investigated the effects of vitamin A and Fe supplementation during gestation on infant mental and psychomotor development. Mothers of infants from five villages in Indonesia were randomly assigned to supervised, double-blind supplementation once per week from approximately 18 weeks of pregnancy until delivery. Supplementation comprised 120 mg Fe+500 microg folic acid with (n 94) or without (n 94) 4800 microg retinol in the form of retinyl acetate. Mothers of infants who participated in the national Fe+folic acid supplementation programme, but whose intake of supplements was not supervised, were recruited from four other villages (n 88). The mental and psychomotor development of infants was assessed, either at 6 or 12 months of age, using the Bayley Scales of Infant Development (BSID). We found no impact of vitamin A supplementation on mental or psychomotor development of infants. In addition, infants whose mothers had received weekly Fe supplementation had similar mental and psychomotor indices as those whose mothers had participated in the governmental Fe supplementation programme. The study population was moderately Fe and vitamin A deficient. The size of the treatment groups was large enough to detect a mean difference of 10 points on the BSID, which is less than 1 sd (15 points) of the average performance of an infant on the BSID. In conclusion, the present study did not find an impact of weekly supplementation of 4800 RE vitamin A in addition to Fe during gestation on functional development of Indonesian infants. However, smaller improvements in development may be seen if studied in a larger and/or more deficient population. PMID- 14756915 TI - Dephytinisation of soyabean protein isolate with low native phytic acid content has limited impact on mineral and trace element absorption in healthy infants. AB - Infant formulas based on soyabean protein isolate are often used as an alternative to cows'-based formulas. However, the presence of phytic acid in soya formulas has raised concern about the absorption of trace elements and minerals from these products. The aim of the present study was to evaluate mineral and trace element absorption from regular and dephytinised soya formula in healthy infants. Soyabean protein isolate with a relatively low native content of phytic acid was used for production of a regular soya formula (300 mg phytic acid/kg liquid formula) and an experimental formula was based on dephytinised soya protein isolate (<6 mg phytic acid/kg liquid formula). Using a crossover study design, apparent mineral and trace element absorptions were measured by a stable isotope technique based on 72 h faecal excretion of non-absorbed stable isotopes (Zn, Fe, Cu and Ca) and by the chemical balance technique (Mn, Zn, Cu and Ca) in nine infants (69-191 d old). Fe absorption was also measured by erythrocyte incorporation 14 d after intake. The results from the present study demonstrated that Zn absorption, measured by a stable isotope technique, was significantly greater after dephytinisation (mean value 16.7 v. 22.6 %; P=0.03). No other statistically significant differences between the two formulas were observed. The nutritional benefit of dephytinisation was marginal in the present study. Based on these results, the use of soyabean protein isolate with low native content of phytic acid should be promoted for production of soya formulas and adequate addition of ascorbic acid to enhance Fe absorption should be ensured in the products. PMID- 14756916 TI - Effect of blood sampling schedule and method of calculating the area under the curve on validity and precision of glycaemic index values. AB - To evaluate the suitability for glycaemic index (GI) calculations of using blood sampling schedules and methods of calculating area under the curve (AUC) different from those recommended, the GI values of five foods were determined by recommended methods (capillary blood glucose measured seven times over 2.0 h) in forty-seven normal subjects and different calculations performed on the same data set. The AUC was calculated in four ways: incremental AUC (iAUC; recommended method), iAUC above the minimum blood glucose value (AUCmin), net AUC (netAUC) and iAUC including area only before the glycaemic response curve cuts the baseline (AUCcut). In addition, iAUC was calculated using four different sets of less than seven blood samples. GI values were derived using each AUC calculation. The mean GI values of the foods varied significantly according to the method of calculating GI. The standard deviation of GI values calculating using iAUC (20.4), was lower than six of the seven other methods, and significantly less (P<0.05) than that using netAUC (24.0). To be a valid index of food glycaemic response independent of subject characteristics, GI values in subjects should not be related to their AUC after oral glucose. However, calculating GI using AUCmin or less than seven blood samples resulted in significant (P<0.05) relationships between GI and mean AUC. It is concluded that, in subjects without diabetes, the recommended blood sampling schedule and method of AUC calculation yields more valid and/or more precise GI values than the seven other methods tested here. The only method whose results agreed reasonably well with the recommended method (ie. within +/-5 %) was AUCcut. PMID- 14756917 TI - Phenolic acid metabolites as biomarkers for tea- and coffee-derived polyphenol exposure in human subjects. AB - Tea and coffee are rich in polyphenols with a variety of biological activities. Many of the demonstrated activities are consistent with favourable effects on the risk of chronic diseases. 4-O-methylgallic acid (4OMGA) and isoferulic acid are potential biomarkers of exposure to polyphenols derived from tea and coffee respectively. 4OMGA is derived from gallic acid in tea, and isoferulic acid is derived from chlorogenic acid in coffee. Our major objective was to explore the relationships of tea and coffee intake with 24 h urinary excretion of 4OMGA and isoferulic acid in human subjects. The relationships of long-term usual (111 participants) and contemporaneously recorded current (344 participants) tea and coffee intake with 24 h urinary excretion of 4OMGA and isoferulic acid were assessed in two populations. 4OMGA was related to usual (r 0.50, P<0.001) and current (r 0.57, P<0.001) tea intake, and isoferulic acid was related to usual (r 0.26, P=0.008) and current (r 0.18, P<0.001) coffee intake. Overall, our present results are consistent with the proposal that 4OMGA is a good biomarker for black tea-derived polyphenol exposure, but isoferulic acid may be of limited usefulness as a biomarker for coffee-derived polyphenol exposure. PMID- 14756918 TI - Factors associated with Hb concentration in children aged 6-59 months in the State of Pernambuco, Brazil. AB - In 1997, the prevalence of anaemia was 40.9 % among children aged 6-59 months in the State of Pernambuco, north-east Brazil. Using the same sample of children, we have investigated possible reasons for this high prevalence. A representative sample was selected through a three-stage process: proportional systematic random sampling of municipalities in the State, systematic random sampling of census sectors within these municipalities, and finally, simple random sampling of households with children aged 6-59 months to obtain the sample of 650 children. Data collection included demographic, environmental, socio-economic and maternal variables, and nutritional status and dietary intakes of the children. Multiple linear regression analysis was based on a hierarchical model of factors associated with Hb concentration. The mean Hb concentration of children aged 6-23 months was 10 g/l lower than that of older children. In the regression analysis, child age explained 8.3 % of the variance in Hb concentration. The intake of bioavailable Fe explained a further 3.3, serum retinol 2.7, diarrhoea 2.4, water treatment 1.7, sanitation 1.3 and low birth-weight 0.5 %. The final model explained 23.4 % of the variance in Hb concentration. We conclude that child age, bioavailable-Fe intake, serum retinol concentration, diarrhoea, water treatment, sanitation and low birth-weight are independently associated with Hb concentration. In north-east Brazil, anaemia prevention programmes among children should focus on those aged <2 years and should consider feasible strategies to improve intakes of bioavailable Fe and vitamin A, and reduce infection. Supplemental Fe should be given to low birth-weight infants. PMID- 14756919 TI - Emerging diet-related surrogate end points for colorectal cancer: UK Food Standards Agency diet and colonic health workshop report. AB - The UK Food Standards Agency convened a group of expert scientists to review current research investigating emerging diet-related surrogate end points for colorectal cancer (CRC). The workshop aimed to overview current research and establish priorities for future research. The workshop considered that the validation of current putative diet-related surrogate end points for CRC and the development of novel ones, particularly in the emerging fields of proteomics, genomics and epigenomics, should be a high priority for future research. PMID- 14756922 TI - Internet gambling: issues, concerns, and recommendations. AB - The influence of technology in the field of gambling innovation continues to grow at a rapid pace. After a brief overview of gambling technologies and deregulation issues, this review examines the impact of technology on gambling by highlighting salient factors in the rise of Internet gambling (i.e., accessibility, affordability, anonymity, convenience, escape immersion/dissociation, disinhibition, event frequency, asociability, interactivity, and simulation). The paper also examines other factors in relation to Internet gambling including the relationship between Internet addiction and Internet gambling addiction. The paper ends by overviewing some of the social issues surrounding Internet gambling (i.e., protection of the vulnerable, Internet gambling in the workplace, electronic cash, and unscrupulous operators). Recommendations for Internet gambling operators are also provided. PMID- 14756923 TI - Pushing the wrong buttons: men's and women's attitudes toward online and offline infidelity. AB - Despite current researchers' interest in the study of online sexual addiction, there is a dearth of research available on what constitutes online infidelity. This paper attempts to redress this balance by comparing 1,117 participants' attitudes toward online and offline acts of infidelity. A factor analysis was carried out that yielded three components of infidelity: sexual infidelity, emotional infidelity, and pornography. More importantly, this study revealed that online acts of betrayal do not fall into a discrete category of their own. A MANOVA was performed and revealed a statistically significant difference on the combined dependent variables for the interaction of gender by age, age by relationship status, and Internet sexual experience. The hypotheses were, in part, supported. However, counter to what was predicted, in the main younger people were more likely to rate sexual acts as acts of betrayal than older individuals. It is concluded here that individuals do perceive some online interactions to be acts of betrayal. In contrast to some researchers' claims, it is suggested here that we do need to consider how bodies are reconstructed online. Moreover, these results have important implications for any treatment rationale for infidelity (both online and offline). PMID- 14756924 TI - Internet users' prior psychological and social difficulties. AB - This study addressed the hypothesis that frequent Internet use produces social and psychological difficulties. An Internet-administered survey was given to a sample of Internet users. Comparisons were made between this sample and general population norms on a selection of social and psychological variables. Internet users showed a more detrimental mean rating on 11 of 13 of the variables (two measures of social contact were more positive among the user sample). However, for those variables for which time of onset was available (n = 7), the disorder had begun 5-22 years before Internet use. These findings are not in accord with the theory that Internet use causes disorder or social difficulty, and suggest the possibility that the Internet may provide a particular benefit for certain individuals who have already displayed these personal and social difficulties. PMID- 14756925 TI - Developing a virtual reality-based methodology for people with dementia: a feasibility study. AB - The aim of this study was to examine the feasibility of virtual reality (VR) technology for use by persons with dementia (PWD). Data were obtained directly from six PWD regarding their experiences with a virtual environment (VE) of a large outdoor park. A user-centered method was developed to assess: (a) presence; (b) user inputs; (c) display quality; (d) simulation fidelity; and (e) overall system usability. The extent to which PWD could perform four functional activities in the VE was also investigated (e.g., mailing a letter). In addition, physical and psychological well-being of PWD while interacting with the VE was assessed objectively by recording heart rate during the VR sessions and subjectively with discrete questionnaire items and real-time prompts. Symptom profiles associated with simulator sickness were assessed with an adapted version of the Simulator Sickness Questionnaire. The study found that PWD to some extent experienced presence; perceived that objects were realistic and moved naturally; generally felt in control of the interaction; and demonstrated little difficulty using a joystick for navigation. The study also demonstrated that VR is an appropriate medium for assessing functional behavior within the context of an ecologically valid VE. PWD did not experience any significant increase in symptoms associated with simulator sickness, or detriments to their psychological and physical well-being. These findings demonstrated that it is feasible to work in VEs with PWD. PMID- 14756926 TI - The world wide web and the laboratory: a comparison using face recognition. AB - In recent years, a growing number of psychological researchers have turned to the World Wide Web (WWW) as a resource to access participants in experimental studies. While there are benefits to this approach in conducting psychological research (e.g., access to a potentially large subject pool and faster data collection), there are also concerns regarding this medium (e.g., the validity of the data). In recent years, data collected on-line has been validated by comparing it to data collected in the traditional laboratory setting. This study attempted to build on these previous reports by comparing face recognition data collected on the web and data collected in a laboratory. In two separate experiments, data collected from WWW participants did not statistically differ from data collected with undergraduate college students in a classroom setting. These findings strongly suggest that the WWW may be a viable alternative for researchers conducting face recognition experiments. PMID- 14756927 TI - Doing play: competency, control, and expression. AB - This qualitative study investigated the personal experiences of children with cerebral palsy engaging in a virtual reality play intervention program. The study involved in-depth, focused interviews that were conducted with 19 participants aged 8-13 who had a diagnosis of cerebral palsy. A constant comparative inductive method of analysis was used, and several themes emerged. Findings were interpreted using the theory of flow, the theory of self-efficacy, and the model of playfulness in virtual reality computer interactions. The three themes uncovered in the data include (a) doing play, (b) it's my way that matters, and (c) how I see me. The sub-theme safety also emerged. Findings from this study showed that children with physical disabilities are often limited in their play experience compared to their peers without physical disabilities. Children perceived engagement in a virtual reality play intervention program as an enjoyable experience which increased their self-competence and self-efficacy. Participants experienced a sense of control and mastery over the virtual environment and were provided a safe way to explore and challenge their abilities. Participants perceived experiencing flow and reported perceived physical changes and increased social acceptance from both peers and family. These findings provide evidence that virtual reality continues to show promise as a pediatric rehabilitation play intervention tool. PMID- 14756928 TI - Virtual reality as a pediatric pain modulation technique: a case study. AB - Post-surgical pain has been consistently reported in pediatrics as being difficult to manage and limiting to surgical outcomes. Pain management of children is not ideal, and some children unable to tolerate traditional pharmacological agents. Virtual reality (VR) is a new and promising form of non pharmacologic analgesia. This case study explored the use of VR analgesia with a 16-year-old patient with cerebral palsy participating in a twice-daily physiotherapy program following Single Event Multi-Level Surgery. Over 6 days, the patient spent half of his physiotherapy sessions using VR and the other half without (order randomized). Traditional pharmacological pain management was administered throughout the trial. Using a subjective pain scale (five faces denoting levels of pain), the patient's overall pain ratings whilst in the VR (experimental) condition were 41.2% less than those in the no-VR (control) condition. This case report provides the first evidence that VR may serve as a powerful non-pharmacologic analgesic for children following surgery. PMID- 14756929 TI - Physical space and cyberspace: how do they interrelate? A study of offline and online social interaction choice in Singapore. AB - This paper investigates if the new space created by cyberspace affects how we use old space--physical space--and how the range of physical space we have access to affects our urge to extend into cyberspace. To do this I polled young IT savvy people in land-scarce Singapore about the amount of physical space they had at their disposal--private space in terms of the size of their house and bedroom space, and public space in terms of how many public meeting areas they frequented. I attempted to see if there was a correlation between the amount of physical space they enjoyed and the amount of time they spent in cyberspace. PMID- 14756930 TI - A magnet-friendly virtual reality fiberoptic image delivery system. AB - A custom display was built into the MR radiofrequency headcoil to project high resolution, wide field-of-view stereographic images. Advanced stimulus presentation technologies such as the one described could potentially contribute to a better understanding of the relation between what people are thinking or experiencing, and their associated patterns of brain activity (www.vrpain.com). PMID- 14756931 TI - Internet addiction of adolescents in Taiwan: an interview study. AB - This study presented a follow-up investigation of a series of studies about Taiwanese adolescents' Internet addiction. Through analyzing a series of questionnaires, 10 Taiwanese adolescents with Internet addiction were selected for in-depth interviews. Adolescents' interview data revealed the following: First, almost all of the interviewed adolescents exhibited most of the symptoms of Internet addiction as identified by previous studies, including compulsive use and withdrawal, tolerance, and related problems of school, health, family, finance, and time management. Moreover, almost all of the interviewed adolescents stated that messages on the Internet became the primary source of their information and knowledge. Many of them were addicted to the messages and activity on the Internet, but not by the Internet as a medium per se. Also, the world of the Internet seemed to become a place to relieve adolescents' depression. Most of the interviewed adolescents self-declared that they were Internet addicts, but none of them could clearly state what to do with such an addiction. Psychologists and educators should pay more attention to this and then propose possible solutions for adolescents. PMID- 14756932 TI - Internet addiction in students: a cause of concern. AB - The Internet was originally designed to facilitate communication and research activities. However, the dramatic increase in the use of the Internet in recent years has led to pathological use (Internet addiction). This study is a preliminary investigation of the extent of Internet addiction in school children 16-18 years old in India. The Davis Online Cognition Scale (DOCS) was used to assess pathological Internet use. On the basis of total scores obtained (N = 100) on the DOCS, two groups were identified--dependents (18) and non-dependents (21), using mean +/- 1/2 SD as the criterion for selection. The UCLA loneliness scale was also administered to the subjects. Significant behavioral and functional usage differences were revealed between the two groups. Dependents were found to delay other work to spend time online, lose sleep due to late-night logons, and feel life would be boring without the Internet. The hours spent on the Internet by dependents were greater than those of non-dependents. On the loneliness measure, significant differences were found between the two groups, with the dependents scoring higher than the non-dependents. PMID- 14756933 TI - Use of virtual reality as a distractor for painful procedures in a patient with pediatric cancer: a case study. AB - Virtual reality (VR) has been demonstrated as an effective tool to help people overcome a variety of anxiety disorders. In this case study, the use of VR as a distractor to alleviate pain and anxiety associated with an invasive medical procedure for a pediatric cancer patient was explored. An A-B-C-A design during four consecutive medical appointments in an outpatient oncology clinic compared no distraction (A), non-VR distraction on a computer screen (B), and VR distraction with a head set (C). Behavioral observations of distress by the researcher and reports of pain and anxiety by the patient, parent, and nurse were taken before and during the procedure. The child's pulse was monitored throughout the procedure. The findings from this case study suggest benefit from using VR distraction, as indicated by lower pain and anxiety ratings, reduced pulse, and fewer observed behavioral indices of distress. The need for larger scale studies and application of VR with younger children is discussed in the context of confirming effectiveness of this technique and providing more generalizable information about efficacy. PMID- 14756934 TI - The role of flow experience in cyber-game addiction. AB - Consumer habit, an important key to repetitive consumption, is an interesting yet puzzling phenomenon. Sometimes this consumption becomes obsessive--consumers will continue to act a certain way even when they feel it is not in their best interests. However, not all consumers develop such addictions. This study uses cyber-game addiction syndrome as an analogue to trace the possible causes of consumer addiction. Results from structure equation modeling show that repetition of favorite activities has a moderate effect upon addiction, which is in line with the assertion of rational addiction theory. However, flow experience--the emotional state embracing perceptional distortion and enjoyment--shows a much stronger impact on addiction. This suggests that consumers who have experienced flow are more likely to be addicted. PMID- 14756935 TI - Determinants of satisfaction with an automated alcohol evaluation program. AB - High rates of untreated mental illness cause serious health problems in the United States and worldwide. The use of computer-administered therapy has the potential to increase access to mental health care for certain patient populations. An online version of an alcohol check-up was developed that guided subjects through a series of standardized questionnaires, and provided them with feedback designed to enhance their appreciation of the negative aspects of their alcohol use. Ratings of the helpfulness of the questionnaires were evaluated in order to determine the characteristics of individuals who would potentially benefit from an automated substance abuse intervention, and to learn which aspects of an automated program would be most useful. Over a period of 25 months, 1,455 individuals participated in the study, 83% of whom had an Alcohol Use Disorders Identification Test (AUDIT) score indicative of problem drinking. Subjects with alcohol problems found the on-line program more useful than other subjects. The questionnaire which compared subjects' alcohol use to national norms, provided the most helpful and also the most unexpected information. Alcohol abusers with high levels of ambivalence about their drinking and other measures of motivation for change found the program more helpful than those with lower motivation. Automated therapy made available to the general public via the Internet can be accessed by large numbers of individuals. The interaction can be useful to patients with significant levels of substance abuse. PMID- 14756938 TI - The personalized consent form: an optional, but useful tool! AB - Dentistry began to incorporate informed consent and risk management into its practice after 1950. Today, an obligation to ensure that the patient has accepted a given treatment and understands all its implications are part of a dentist"s prime responsibilities. The purpose of this article is to inform dentists who are asking for consent that they can use a tool to make this task easier--the personalized consent form. PMID- 14756939 TI - Smoking cessation services provided by dental professionals in a rural Ontario health unit. AB - PURPOSE: This study was undertaken to determine what smoking cessation services dental professionals in Ontario's Wellington-Dufferin-Guelph Health Unit (WDGHU) provide before disseminating a smoking cessation information package. METHODS: Data were collected with 540 self-administered questionnaires mailed to 60 local dental offices. Replies were requested from all dentists, dental hygienists, dental assistants and other dental staff working in each dental office. RESULTS: Completed responses were obtained from 126 dental personnel in 28 (47%) of the 60 dental offices surveyed. The proportion of dental offices, dentists and hygienists providing cessation services to most patients was as follows: asking patients about tobacco-use status, 46%, 31% and 32%; advising tobacco users to quit, 46%, 32% and 29%; assessing tobacco users' interest in quitting, 46%, 25% and 19%; and assisting interested patients to quit, 25%, 6% and 13%, respectively. CONCLUSION: This survey indicates that most dental professionals in the WDGHU do not provide proven smoking cessation services. An opportunity exists to increase the proportion of dental professionals providing proven smoking cessation interventions as part of routine patient services. PMID- 14756940 TI - Recurrent desmoplastic ameloblastoma of the maxilla: a case report. AB - A case of desmoplastic ameloblastoma recurring within 2 months of curettage is presented. This tumour appeared in the premolar region of the left maxilla with involvement of the antrum. The 24-year-old female patient was initially treated by curettage with wide surgical margins. Later, partial maxillectomy was carried out followed immediately by iliac bone graft. The case was followed with periodic plain radiography and computed tomography. The presence of a pulpally infected premolar and the atypical radiographic appearance obscured the disease. The biologic profile of this tumour is not fully understood because of the limited number of reported cases, coupled with inadequate long-term follow-up. A review of the lesion with emphasis on the pathogenesis of recurrence is discussed. PMID- 14756941 TI - Effect of disposable infection control barriers on light output from dental curing lights. AB - PURPOSE: To prevent contamination of the light guide on a dental curing light, barriers such as disposable plastic wrap or covers may be used. This study compared the effect of 3 disposable barriers on the spectral output and power density from a curing light. The hypothesis was that none of the barriers would have a significant clinical effect on the spectral output or the power density from the curing light. METHODS: Three disposable barriers were tested against a control (no barrier). The spectra and power from the curing light were measured with a spectrometer attached to an integrating sphere. The measurements were repeated on 10 separate occasions in a random sequence for each barrier. RESULTS: Analysis of variance (ANOVA) followed by Fisher's protected least significant difference test showed that the power density was significantly less than control (by 2.4% to 6.1%) when 2 commercially available disposable barriers were used (p < 0.05). There was no significant difference in the power density when general purpose plastic wrap was used (p > 0.05). The effect of each of the barriers on the power output was small and probably clinically insignificant. ANOVA comparisons of mean peak wavelength values indicated that none of the barriers produced a significant shift in the spectral output relative to the control ( p > 0.05). CONCLUSIONS: Two of the 3 disposable barriers produced a significant reduction in power density from the curing light. This drop in power was small and would probably not adversely affect the curing of composite resin. None of the barriers acted as light filters. PMID- 14756942 TI - [Broaden the scope of study on gastrointestinal diseases and improve the levels of diagnosis and treatment]. PMID- 14756943 TI - [Biological characteristics of influenza virus]. PMID- 14756944 TI - [Diagnosis and treatment of intestinal stone obstruction in infants by combined use of ultrathin gastroscopy and enteroscopy]. PMID- 14756945 TI - [Levels and mRNA expression of interleukin-8 in gastric and duodenal mucosa of children Infected with Helicobacter pylori]. AB - OBJECTIVE: To determine the changes of IL-8 and IL-8 mRNA in gastric and duodenal mucosa of children with Hp infection, to study the effect of Helicobacter pylori (Hp) infection on the expression of IL-8 and IL-8 mRNA, and to evaluate its possible roles in the pathogenesis of gastric and duodenal mucosal inflammation in Hp related gastroduodenal diseases. METHODS: Biopsy specimens were taken from the antral and duodenal mucosa on endoscopy in patients with or without Hp infection, which was diagnosed by urease test and Warshing-Starry staining. The expression of IL-8 in gastric and duodenal mucosa was determined by ELISA, the expression of IL-8 mRNA was determined by using RT-PCR. RESULTS: Inflammation of gastric antral mucosa was more severe in Hp-positive group than in Hp-negative group. Active inflammation often existed on the basis of chronic inflammation in Hp-positive mucosa, and duodenal mucosa had mild chronic inflammation in Hp positive group. Of 17 children who were not infected with Hp, 4 had pathologically normal gastric mucosa and had mild chronic gastritis, one child had an active chronic gastritis. Nineteen children were infected with Hp and all had chronic gastritis with signs of active inflammation. Gastric and duodenal mucosal IL-8 and IL-8 mRNA were higher in HP infected than in non infected children (IL-8: in gastric mucosa 24.66 - 177.77 pg/mg, 2.94 - 12.98 pg/mg, t = 12.34, P < 0.01; in duodenal mucosa: 4.28 - 47.76 pg/mg, 2.04 - 9.52 pg/mg, t = 7.18, P < 0.01. IL-8 mRNA: in gastric mucosa 2.37 - 4.99, 0.05 - 0.44, t = 29.29, P < 0.01; in duodenal mucosa 1.22 - 1.87, 0.01 - 0.23, t = 37.20, P < 0.01). Children with active chronic gastritis had higher interleukin-8 levels and IL-8 mRNA expression than those with inactive gastritis (IL-8 in gastric mucosa: 12.98 - 177.77 pg/mg, 2.04 - 10.43 pg/mg, t = 10.66, P < 0.01; in duodenal mucosa: 5.28 - 47.76 pg/mg, 3.19 - 8.14 pg/mg, t = 6.52, P < 0.01. IL-8 mRNA in gastric mucosa: 0.51 - 4.99, 0.01 - 0.44, t = 18.62, P < 0.01; in duodenal mucosa: 0.23 - 1.87, 0.01 - 0.20, t = 19.10, P < 0.01). CONCLUSION: Higher expression of IL-8 and IL-8 mRNA was seen in Hp-positive gastric and duodenal mucosa and in active gastritis. IL-8 may play an important role in the local gastric and duodenal mucosal inflammatory infiltration with a large number of neutrophils when there is Hp infection. PMID- 14756946 TI - [Report of 3 cases with severe adenoviral pneumonia in infants and young children]. PMID- 14756947 TI - [Effect of Helicobacter pylori infection on iron status among preschool children]. AB - OBJECTIVE: To explore the relationship between Helicobacter pylori (Hp) infection and iron status using serum ferritin (SF) as a marker for total iron and to identify the related factors of iron nutritional status among preschool children. METHODS: By cluster sampling, we recruited 475 preschool children aged 2 to 7 years. A structured questionnaire and diet form were sent to the parents of these children to obtain related information about the socioeconomic level and dietary intakes. After collecting blood samples, the following indexes were measured. Hp IgG antibodies were measured with a dot enzyme-linked immunoassay; hemoglobin, Hct, mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), red blood cell distribution width index (RDW) with automatic Complete Blood Count; SF with an immunoradiometric assay. Stool Hp antigen and occult bleeding were measured with ELISA among individuals who were Hp seropositive. Hp status was defined as positive when both serum and stool antigen tests were positive, Hp status was defined as negative when serum antigen test was negative; 24-hour weighting and recording methods were used to dietary survey for three days in May and December 2001, respectively, dietary intakes including energy, protein and micronutrient were calculated using nursery school nutrition software and evaluated by Chinese Dietary Reference Intakes (DRIs). Mann-Whitney test was used to compare mean ranks of SF in Hp-positive and Hp-negative children. To obtain an adjusted estimate of the impact of Hp infection on SF, a multivariate analysis of covariance was done to evaluate the different level of SF between Hp infected and non-infected status. The relationship between iron deficiency and gender, age, socioeconomic condition, iron intake, and calcium intake was assessed by univariate analysis. An unconditional multivariate logistic regression analysis was also performed. Iron deficiency status was dichotomized and placed as the dependent variable. Hp infection status was incorporated together with possible confounding factors as independent variables in a final logistic regression model. All the data were managed by EPI Info 5.01a and analyzed by SAS (Version 6.12). RESULTS: Totally 64 children were diagnosed as Hp-positive and 305 as Hp negative. Mann-Whitney test and multivariate analysis of covariance both showed that SF concentration was significantly lower in Hp infected individuals than non infected individuals. Adjusted mean level and 95% confidence interval of SF in infected and non-infected children was 23.62 microg/L (7.13 microg/L-78.26 microg/L), 33.48 microg/L (10.28 microg/L-109.06 microg/L), respectively. The relationship between Hp infection and iron deficiency status persisted in logistic regression analysis after adjusting for possible confounding factors (OR: 7.95; 95% CI 2.56 - 24.67). CONCLUSION: Iron nutritional status was reduced in Hp infected preschool children. Hp infection appears to be an independent risk factor or an added stressor on iron status among preschool children. PMID- 14756948 TI - [Determination of total and segmental colonic transit time in constipated children]. AB - OBJECTIVE: To determine the total and segmental colonic transit time of normal Chinese children and to explore its value in constipation in children. METHODS: The subjects involved in this study were divided into 2 groups. One group was control, which had 33 healthy children (21 males and 12 females) aged 2 - 13 years (mean 5 years). The other was constipation group, which had 25 patients (15 males and 10 females) aged 3 - 14 years (mean 7 years) with constipation according to Benninga's criteria. Written informed consent was obtained from the parents of each subject. In this study the simplified method of radio opaque markers was used to determine the total gastrointestinal transit time and segmental colonic transit time of the normal and constipated children, and in part of these patients X-ray defecography was also used. RESULTS: The total gastrointestinal transit time (TGITT), right colonic transit time (RCTT), left colonic transit time (LCTT) and rectosigmoid colonic transit time (RSTT) of the normal children were 28.7 +/- 7.7 h, 7.5 +/- 3.2 h, 6.5 +/- 3.8 h and 13.4 +/- 5.6 h, respectively. In the constipated children, the TGITT, LCTT and RSTT were significantly longer than those in controls (92.2 +/- 55.5 h vs 28.7 +/- 7.7 h, P < 0.001; 16.9 +/- 12.6 h vs 6.5 +/- 3.8 h, P < 0.01; 61.5 +/- 29.0 h vs 13.4 +/- 5.6 h, P < 0.001), while the RCTT had no significant difference. X-ray defecography demonstrated one rectocele, one perineal descent syndrome and one puborectal muscle syndrome, respectively. CONCLUSION: The TGITT, RCTT, LCTT and RSTT of the normal children were 28.7 +/- 7.7 h, 7.5 +/- 3.2 h, 6.5 +/- 3.8 h and 13.4 +/- 5.6 h, respectively. With the segmental colonic transit time, constipation can be divided into four types: slow-transit constipation, outlet obstruction, mixed type and normal transit constipation. X-ray defecography can demonstrate the anatomical or dynamic abnormalities within the anorectal area, with which constipation can be further divided into different subtypes, and combined use of the gastrointestinal transit time and X-ray defecography is of clinical importance in exploration of etiology of constipation. PMID- 14756949 TI - [Blood motilin concentration and enteral nutrition in premature infants]. AB - OBJECTIVE: To study changes of plasma motilin concentration and it's effect on enteral nutrition in premature infants. METHODS: The plasma motilin concentration of 72 premature infants was measured within 12 hours after birth before enteral feeding and on day 3 and 7 of life by using radioimmunoassay. Sixteen full-term neonates were enrolled as controls. RESULTS: (1) The plasma concentrations of motilin in premature infants before enteral feeding after birth and on day 3 and 7 were 198.65 +/- 58.42 ng/L, 248.83 +/- 56.00 ng/L, and 376.77 +/- 139.46 ng/L, respectively, which were significantly lower than those in the control group (300.33 +/- 67.15 ng/L, 334.26 +/- 83.81 ng/L, 510.64 +/- 179.85 ng/L) (P < 0.001 or < 0.01). There was positive correlation between the concentration and gestational age, age in day and the volume of milk. On day 7 the level of motilin was higher than the pre-enteral feeding level of the full term control group. (2) The plasma motilin concentration in feeding un-tolerated premature infants group was lower than that in the normal group, especially on day 3 of life (P < 0.05). (3) Early enteral feeding could improve the plasma motilin levels, gastrointestinal motility and nutrition tolerance in premature infants. CONCLUSIONS: The gastrointestinal functions of premature infants are adaptable to enteral nutrition. Early enteral feeding (including minimal enteral nutrition and non-nutritive sucking) can promote adaptive rapid growth and development of intestine. PMID- 14756950 TI - [A case with Gilles de la Tourette syndrome associated with depression]. PMID- 14756951 TI - [Summary of the 4th national symposium on childhood gastrointestinal diseases]. PMID- 14756952 TI - [Summary of China National Pediatric Hematology Conference (2002)]. PMID- 14756953 TI - [Routine methods for cleaning and sterilization of gastrointestinal endoscopes]. PMID- 14756954 TI - [Gastroscopic diagnostic criteria for chronic gastritis and peptic ulcer in children]. PMID- 14756955 TI - [The recommended protocols for pharmacotherapy of peptic ulcer in children]. PMID- 14756956 TI - [Progresses in studies on childhood functional constipation]. PMID- 14756957 TI - [Early colonization of normal microflora in oral cavity of children]. PMID- 14756958 TI - [Prediction of walking and life span of children with cerebral palsy]. PMID- 14756959 TI - [Analysis of 187 children with enteroviral central nervous system infection in Shandong area]. AB - OBJECTIVE: To evaluate the diagnostic potential of previously published enterovirus (EV) reverse transcription polymerase chain reaction (RT-PCR) assay in detection of EV in CSF samples from children with a diagnosis of aseptic meningitis and to investigate the clinical characteristics of the patients seen in Shandong. METHODS: EV RNA was detected in 187 CSF samples and serum and/or urine samples of a part of patients by RT-PCR and viral culture technique. RESULTS: RT-PCR was positive in all 62 CSF specimens which were positive by cell culture (100%). In addition, 93 of 125 (74.4%) CSF samples negative by cell culture were RT-PCR positive. In 4 of these 93 (4.3%) patients, viral culture of specimens from other sites (serum or urine) was also positive. The sensitivity of CSF RT-PCR based on clinical diagnosis in patients with meningitis of negative bacterial culture results was 82.9% (155/187), which was considerably higher than the sensitivity of CSF virus culture 33.2% (62/187). The results of RT-PCR can be reported within 4 hours, whereas the viral culture of CSF requires 4.6 days for a cytopathic effect to develop. EV meningitis occurred in a sporadic form and in some areas there were outbreaks. The clinical characteristics of 155 patients with EV meningitis were different in different age groups. CONCLUSION: EV was one of the most common causes of aseptic meningitis in Shandong area. The RT-PCR assay was rapid, sensitive and specific for the diagnosis of EV meningitis and may be a potential tests to shorten hospital stay and reduce the use of antibiotics. PMID- 14756960 TI - [Clinical characteristics of X-linked adrenoleukodystrophy]. AB - OBJECTIVE: X-linked adrenoleukodystrophy (ALD) is a genetically determined disorder that involves the nervous system white matter, axons, adrenal cortex and testes. The typical clinical manifestations are progressive psychomotor regression, vision and/or auditory impairment and adrenal insufficiency. The clinical manifestation, biochemical change and genetic counseling work of X linked ALD were analyzed. METHODS: The clinical features of 29 cases with ALD were summarized and analyzed, including symptoms and signs, measurement of blood very long chain fatty acids (VLCFA), adrenal function, cranial magnetic resonance imaging (MRI) and pedigree investigation. RESULTS: Among these 29 cases, the clinical phenotype could be classified into childhood cerebral (22 cases), adolescent cerebral (4 cases), adrenomyeloneuropathic (1 case), Addison's disease (1 case) and asymptomatic or presymptomatic (1 case) types. Nine of them had positive family history. Pedigree investigation was consistent with typical sex linked recessive inheritance. There were 45 ALD patients in these 29 pedigrees. The neurological manifestations varied among members of the same family. Nine cases died during follow up. The causes of death were central respiratory failure or other complications of ALD and so on. Laboratory tests demonstrated abnormally high plasma levels of VLCFA in ALD patients; MRI demonstrated symmetric butterfly like low T(1) and high T(2) signals in the parieto-occipital white matter. The impairment in the splenium of corpus callosum made the bilateral lesion region converge into one. It could progress anteriorly and injure the bilateral posterior limb of internal capsule and the temporal lobe, and could injure the brainstem inferiorly. Following intravenous injection of contrast material, thin stripe of lacelike enhancement could be observed. CONCLUSIONS: The atypical initial symptom of ALD was seizures. The MRI showed abnormal signal in the cerebellar white matter. This disease can influence the normal development of children, this was more pronounced in the childhood cerebral ALD type. It tended to progress rapidly with dementia, vegetative state or death. Since antenatal diagnostic method is available now, emphasis should be made on the antenatal examination in order to make an early diagnosis and abort pregnancy if necessary. PMID- 14756961 TI - [Effect of intrauterine acute ischemic-hypoxia on the expression of lung SP-A and SP-B in neonatal rats]. AB - OBJECTIVE: Neonatal asphyxia is one of the main causes for the acute respiratory distress syndrome (ARDS) in full-term newborns. Now it is believed that the reduced amount and abnormal function of pulmonary surfactant due to various causes is a major factor leading to acute lung injury. This study aimed at using an intrauterine acute ischemic-hypoxia rat model and investigating the effect of intrauterine acute ischemic-hypoxia on the expression of surfactant protein A (SP A) and surfactant protein B (SP-B) in neonatal rat lungs. METHODS: The rat model of acute intrauterine ischemic-hypoxia was established by ligating the unilateral uterine horn vessels of Wistar rats at the 21st gestational day. While the rat pups from the other side of the uterus, of which the uterine horn vessel was not ligated, were the sham-operation group. Rat pups were delivered by cesarean section at the 20, 30 and 40 min following the ischemic-hypoxia insult. The rat pups delivered by cesarean section from the gestation of 21 days were the normal control group. There were 42 rat pups and 6 pups in each group in this study. The distribution of SP-B protein in the neonatal rat lungs of different period of ischemia was examined by using SABC method. The average gray value of SP-B staining in type II alveolar epithelial cells were measured by Universal Imaging Porporation with Meta Morph software. The reverse transcription polymerase chain reaction (RT-PCR) was performed to quantitate the expression of SP-A and SP-B mRNA. RESULTS: Following the intrauterine acute ischemic-hypoxia, the numbers of type II alveolar epithelial cells with the positive SP-B staining were markedly declined. The average gray values at the 20, 30 and 40 min after the ischemia were 78.89 +/- 1.08, 79.69 +/- 0.13 and 80.00 +/- 0.63, respectively, which increased significantly compared with the normal control group (76.13 +/- 0.43, P < 0.01). The expression of SP-A and SP-B mRNA was weak following the ischemic hypoxia insult. The relative amounts of SP-A (1.16 +/- 0.06, 1.14 +/- 0.01 and 1.13 +/- 0.04, respectively) and SP-B (0.81 +/- 0.02, 0.78 +/- 0.02 and 0.79 +/- 0.04, respectively) at the 20, 30 and 40 min after the ischemia were reduced significantly compared with controls (1.27 +/- 0.09 and 0.89 +/- 0.06, respectively, P < 0.05 and < 0.01) and reduced gradually following the prolongation of the insult. There were no significant differences (P > 0.05) between the normal and sham operation control groups on the expressions of SP-B protein as well as the SP-A and SP-B mRNA. CONCLUSION: The reduced synthesis of SP-B protein and the reduced expression of SP-A and SP-B mRNA might be caused by intrauterine acute ischemic-hypoxia, which may support theoretically the early application of pulmonary surfactant including SP-A and SP-B for treating the lung injuries of asphyxia in newborns. PMID- 14756963 TI - [Endogenous nitric oxide pathway in high pulmonary blood flow-induced pulmonary vascular structural remodeling]. AB - OBJECTIVE: Pulmonary vascular structural remodeling induced by high pulmonary blood flow is an important pathologic basis of pulmonary hypertension with congenital heart disease of left-to-right shunt. However, the mechanism is still not clear. The present study aimed to examine the alteration of endogenous nitric oxide (NO) pathway in high pulmonary blood flow-induced pulmonary vascular structural remodeling, so as to explore the role of NO pathway in pulmonary hypertension induced by high pulmonary blood flow. METHODS: Sixteen male SD rats were randomly divided into control group (n = 8) and shunting group (n = 8). Aortocaval shunting was produced for 11 weeks in shunt rats. Pulmonary artery mean pressure (mPAP) of each rat was evaluated using right cardiac catheterization. The ratio of right ventricular mass to left ventricular plus septal mass [RV/(LV + S)] was detected. Pulmonary vascular micro-and ultra structure was examined by using a light microscope and a transmitted electronic microscope. Meanwhile, the concentration of plasma NO was measured by spectrophotometry. The expressions of endothelial NO synthase (eNOS) mRNA and protein by pulmonary arteries were detected by in situ hybridization and immunohistochemistry, respectively. RESULTS: After 11-week aortocaval shunting, mPAP was significantly increased [(22.5 +/- 2.6) mmHg vs. (15.8 +/- 2.8) mmHg, 1 mmHg = 0.133 kPa, t = 4.97, P < 0.01], and RV/(LV + S) was also markedly increased (0.267 +/- 0.022 vs. 0.221 +/- 0.016, t = 4.85, P < 0.01). The percentage of muscularized arteries was obviously increased in shunt rats compared with controls [(23.2 +/- 2.4)% vs. (13.5 +/- 2.1)%, t = 7.82, P < 0.01], and relative medial thickness of pulmonary arteries was obviously increased in shunt rats [median pulmonary artery: (7.76 +/- 0.56)% vs. (4.82 +/- 1.03)%, t = 6.23, P < 0.01; small pulmonary artery: (11.94 +/- 0.66)% vs. (6.91 +/- 0.53)%, t = 14.96, P < 0.01]. Ultrastructural changes, such as hyperplasia and degeneration of endothelial cells, irregularity of internal elastic laminar and hypertrophy and the increased number of synthetic phenotype of smooth muscle cells, were found in intrapulmonary arteries of shunt rats. Meanwhile, plasma NO concentration was increased [(30.2 +/- 7.9) micromol/L vs (19.7 +/- 5.7) micromol/L, t = 3.05, P < 0.01) and eNOS mRNA and protein expressions by pulmonary arteries were significantly augmented in rats of shunting group. CONCLUSION: The upregulation of eNOS/NO might be an adaptive response of pulmonary circulation to an increased blood flow in the development of pulmonary hypertension and pulmonary vascular structural remodeling. PMID- 14756962 TI - [Expression of caspase-3 mRNA in the hippocampus of seven-day-old hypoxic ischemic rats and the mechanism of neural protection with magnesium sulfate]. AB - OBJECTIVE: There was consanguineous relationship between caspase-3 and early damage after hypoxia and ischemia. Caspase-3 plays a key role in the process of apoptosis in neuron. Magnesium sulfate could protect neuron from injuries, but the mechanism was not clear. The study was to investigate the expression of caspase-3 mRNA in the hippocampus of seven-day-old hypoxic-ischemic rats and the possible mechanism of neural protection with magnesium sulfate. METHODS: The model of seven-day-old hypoxia-ischemia rats was established. The rats were divided randomly into 6 groups as follows: (1) normal control (n = 4); (2) sham surgery control (n = 4); (3) hypoxia-ischemia (n = 4); (4) sodium chloride injection with hypoxia-ischemia (n = 4); (5) magnesium sulfate pre-injection with hypoxia-ischemia (n = 4); (6)magnesium sulfate post-injection with hypoxia ischemia (n = 4). The therapy groups received a bolus injection of 500 mg/kg magnesium sulfate intraperitoneally 0.5 hour before or after hypoxia-ischemia. Semi-quantitative RT-PCR was used to measure caspase-3 mRNA expression in the hippocampus 24 hours after hypoxia-ischemia. RESULTS: The expression of caspase-3 mRNA was significantly increased in the hippocampus of the hypoxia-ischemia pups (1.88 +/- 0.36 vs 0.97 +/- 0.46, P < 0.05). The expression of caspase-3 mRNA in rats with magnesium sulfate pre-injection and post-injection decreased significantly (1.54 +/- 0.49, 1.65 +/- 0.48 vs 1.88 +/- 0.36, P < 0.05). CONCLUSION: Caspase-3 was activated in the hippocampus of the seven-day-old rats 24 hours after hypoxia-ischemia. The suppression of the expression of caspase-3 mRNA in the hippocampus was probably related to the protective effect of magnesium sulfate on the brain injury of hypoxia-ischemia. PMID- 14756964 TI - [Effects of sympathectomy on the blood pressure of young Wistar rats fed with high-sucrose diet]. AB - OBJECTIVE: Diet is an important factor influencing blood pressure and, increases in dietary carbohydrate intake can raise blood pressure in adult rats. A previous study showed that the blood pressure of the rats fed with high-carbohydrate was 5 20 mmHg higher than that of control rats. While the mechanism involved is not clear. This study aimed to investigate the effects of high-sucrose intake on blood pressure of young Wistar rats and the role that sympathetic nerve system in the process. METHODS: Male neonatal Wistar rats were performed sympathectomy operation with 6-hydroxydopamine (6-OHDA) and then divided into four groups: (1) 0.1% VitC saline-common diet group (VN), (2) 0.1% VitC saline-high sucrose (VS), (3) 6-OHDA-common diet group (OHN) and (4) 6-OHDA-high sucrose (OHS) after three week. The data on the body weight (BW), systolic blood pressure (SBP) and heart rate (HR) were recorded. Then the level of blood glucose, serum insulin and angiotensin II (AngII) were measured and the functional studies of the thoracic aorta was performed. RESULTS: The VS group exhibited higher SBP than the OHS group from the 6th week (113.7 +/- 4.2 mmHg vs. 104.0 +/- 5.8 mmHg, P < 0.01) and the VN group from the 7th week (117.6 +/- 6.3 mmHg vs. 109.6 +/- 4.6 mmHg, P < 0.01), while the SBP of the VN group was similar to those of the OHN group and the OHS group (P > 0.05). No significant differences in blood glucose, serum insulin and insulin sensitive index (ISI) were found among the four groups. The thoracic aorta segments of the VS group had higher contractive response to AngII (P < 0.01) and NE (P < 0.05) than the VN group, but the relaxations to acetylcholine (ACh) and nitroglycerine (NTG) showed no difference among the four groups (P > 0.05). CONCLUSION: The high-sucrose diet might elevate the blood pressure in young Wistar rats and the sympathetic system may play an important role in this process. PMID- 14756965 TI - [Analysis of gene mutation in patients with tuberous sclerosis complex with polymerase chain reaction-single strand conformation polymorphism]. AB - OBJECTIVE: Tuberous sclerosis complex (TSC) is an autosomal dominant disease characterized by unusual tumor-like growth, termed hamartomas that develop in a variety of tissues and organs. Clinical findings characteristic of TSC include facial angiofibroma, epilepsy and mental retardation. In the last decade, two genes (TSC1 and TSC2) responsible for this disease were identified and both of them are speculated to be a kind of tumor suppressor gene. TSC1 and TSC2 are located on 9q34 and 16p13.3, respectively. This study was designed to detect gene mutations in patients with TSC. METHODS: All the exons of TSC1 and TSC2 were analyzed by using polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) in DNA separated from peripheral blood of 28 patients with TSC and 100 normal controls. Of the 28 patients, 17 were male and 11 were female, the age of the patients was 1 - 48 years. RESULTS: The mutations were not clustered on a particular exon in either of the genes. Four TSC1 mutations found in 28 cases were on exons (1 nonsense, 2 missense and 1 frameshift); 13 mutations were found in TSC2 gene (2 nonsense, 2 frameshift, 1 deletion and 8 missense). Both TSC1 and TSC2 mutations were detected in 2 cases respectively. The same missense mutation (Q654E) was found in 2 unrelated patients. There was no obvious relationship between the location of the mutation and the clinical symptoms. CONCLUSION: Mutations found in this study were distributed on various exons and there was no clustering of the mutations, the widespread distribution of TSC1/TSC2 mutations hinders the development of a simple diagnostic test, and the identification of individual mutations does not provide prediction of prognosis. PMID- 14756966 TI - [An experimental study on the dilatation of coronary arteries caused by immune vasculitis]. PMID- 14756967 TI - [Diagnostic value of integral of dorsal acoustic scattering for acute viral myocarditis]. PMID- 14756968 TI - [Compared clinical study on treatment of childhood epilepsy with different escalating dosage protocol of topiramate]. PMID- 14756969 TI - [Transcatheter closure of Swiss cheese like ventricular septal defects in children using the Amplatzer muscular VSD occluder]. PMID- 14756970 TI - [Case no. 107--Onset with fever, drowsiness and convulsion]. PMID- 14756971 TI - [A case with giant gastric mucosal hypertrophy]. PMID- 14756972 TI - [Two cases of autoimmune polyadenosis syndrome]. PMID- 14756973 TI - [A case with disseminated eosinophilic fasciitis and myositis]. PMID- 14756976 TI - Associations between perceived parent behaviors and middle school student fruit and vegetable consumption. AB - OBJECTIVE: To examine whether (1) student perceptions of parent behaviors explain variations in fruit and vegetable consumption, (2) self-efficacy mediates this relationship, and (3) perceived home fruit and vegetable availability moderates this relationship. DESIGN: A cross-sectional survey. SETTING: Classrooms in 3 middle schools in 2 northeast Georgia counties. PARTICIPANTS: 366 middle school students. The response and participation rates were 59% and 56%, respectively. VARIABLES MEASURED: Perceived authoritative parenting, perceived parent control, perceived parent modeling, perceived parent support, self-efficacy, perceived fruit and vegetable availability, and fruit and vegetable consumption. ANALYSIS: Hierarchical multiple regression; P <.05. RESULTS: Perceived parent modeling, perceived parent support, self-efficacy, and perceived fruit and vegetable availability were significant predictors of fruit and vegetable consumption. The relationship between perceived parent support and fruit and vegetable consumption was mediated by self-efficacy. The relationship between fruit and vegetable consumption and both perceived parent modeling and support was moderated by perceived fruit and vegetable availability. CONCLUSIONS AND IMPLICATIONS: Parents appear to moderately influence middle school student fruit and vegetable consumption. Educators might focus on improving home fruit and vegetable availability and student self-efficacy, as well as parent support and modeling. The level of availability might indicate where efforts should focus for enhancing parent behaviors. PMID- 14756977 TI - Qualitative study of spirituality in a weight loss program: contribution to self efficacy and locus of control. AB - OBJECTIVE: The purpose of this qualitative study was to examine how spirituality affects intrapersonal characteristics associated with a weight loss program. DESIGN: A series of 5 focus group interviews was conducted with women who were past participants of the Weigh Down Workshop, a spiritually based weight loss program. SETTING: Three churches in the Minneapolis/St. Paul, Minnesota, metropolitan area. PARTICIPANTS: Focus group participants (N = 32) were white, fairly well educated, with moderate income levels. Their mean age was 50 years. PHENOMENA OF INTEREST: Behavior changes, factors affecting self-efficacy for performing the behaviors, and locus of control. ANALYSIS: Sessions were audiotaped and transcribed. Transcribed text was coded and analyzed using qualitative data analysis procedures. RESULTS: Major changes in self-reported eating behaviors included eating only when experiencing true physiological hunger and stopping when sensing a feeling of fullness. Self-efficacy for these behaviors was reported to be enhanced by observing weight loss for themselves or others. Support from other group members, the simplicity of the program, and spiritual benefits through prayer and scripture reading were also reported to enhance confidence. Women indicated that they relied on an internal locus of control based on a sense of self-discipline. CONCLUSIONS AND IMPLICATIONS: Traditional means to enhance self-efficacy were important for all women; however, for some women, spirituality was also an important aspect of adhering to program principles. PMID- 14756978 TI - Pilot test of a behavioral skill building intervention to improve overall diet quality. AB - OBJECTIVE: To determine the effect of a cognitive and behavioral skills building intervention delivered via a small group or correspondence on improvement in total diet quality. DESIGN: Randomized, controlled trial comparing 2 intervention groups with a usual care (UC) group. PARTICIPANTS: Generally healthy men (n = 35) and women (n = 63); mean age = 49.6 years (range = 29 to 71 years). INTERVENTION: 20-session behavioral and cognitive skills curriculum to train participants to improve personal dietary habits that were inconsistent with public health guidelines. One group (weekly meeting [WM]) met in small groups with 2 cofacilitators. A correspondent (CR) group received the curriculum via mail and an interactive study Web site. The UC group received a copy of a consumer nutrition book. MAIN OUTCOME MEASURE: Modified Healthy Eating Index (MHEI) score derived from 9 components of the US Department of Agriculture's Healthy Eating Index. RESULTS: The WM group significantly improved their MHEI score compared with the CR (P =.04) and UC (P =.002) groups. The CR group's improvement in MHEI score was not significantly different from that of the UC group (P =.19). CONCLUSIONS AND IMPLICATIONS: A behaviorally focused intervention can improve overall diet quality, especially if delivered through small-group meetings. PMID- 14756979 TI - Development of an instrument to assess predisposing, enabling, and reinforcing constructs associated with fat intake behaviors of low-income mothers. AB - OBJECTIVE: To establish reliability and construct and discriminant validity of selected factors adapted from the Health and Taste Attitudes Questionnaire developed in Finland and the Food Choice Questionnaire developed in England for use with low-income mothers in the United States. DESIGN: Cross-sectional. SETTING: Head Start and the Special Supplemental Nutrition Program for Women, Infants and Children sites in south-central Wisconsin. PARTICIPANTS: A convenience sample of 211 nonpregnant non-Hispanic black and white women. VARIABLES MEASURED: Predisposing, enabling, and reinforcing constructs of the PRECEDE-PROCEED model. ANALYSIS: Factor analyses. RESULTS: After modification, the resulting measurement models of predisposing, enabling, and reinforcing constructs demonstrated good reliability and construct and discriminant validity. These models showed excellent model fit (chi-square/df range 1.5-2.0, root mean square error of approximate range.05-.07, non-normed fit index range.97-1.0, comparative fit index range.98-1.0, incremental fit index range.98-1.0), with all factor loadings significant (P <.001). Items previously developed with European adults demonstrated different pattern structures in predisposing and enabling constructs but the same structures for reinforcing construct when applied to low income mothers in the United States. CONCLUSION AND IMPLICATION: This modified instrument provides a tool for researchers and educators to further explore factors affecting fat intake behaviors in low-income mothers. PMID- 14756980 TI - Composite index for aggregating nutrient density using food labels: ratio of recommended to restricted food components. AB - This article describes a ratio of recommended to restricted food components (RRR) designed to provide consumers with a summary of food label information to guide healthful, single-item food selections. As a ratio, RRR is interpreted such that better foods score over 1.0. The potential usefulness of the ratio is illustrated comparing foods within categories of the Food Guide Pyramid (eg, skim milk and whole milk). The RRR is proposed for use at the point of purchase for single food items, summarizing the food label, or providing concise information where none is currently presented, such as in restaurants. PMID- 14756981 TI - Is one theory better than another in nutrition education? A viewpoint: more is better. AB - Health behavior theories describe the relations among variables influencing a behavior and specify targets for facilitating behavior change. Nutrition education does not have a dominant theory specific to the discipline. Instead, constructs from multiple theories have been borrowed, primarily from the social sciences, and have been applied to describe or predict nutrition-related behaviors. However, current theories do not fully predict behavior or behavior change. A more effective approach may be to integrate distinct constructs from competing theories into one or more polytheoretical models that can be empirically tested and refined into a more comprehensive, tailored theory or set of theories specific to food and nutrition behavior changes. In our view, more than one will be needed to address the complex array of people, issues, and contexts that we routinely address in nutrition education and behavioral interventions. PMID- 14756982 TI - EatFit: a goal-oriented intervention that challenges adolescents to improve their eating and fitness choices. PMID- 14756983 TI - Barbershop nutrition education. PMID- 14756985 TI - [Outpatient care or hospitalization? A crucial decision in the treatment of community-acquired pneumonia]. PMID- 14756986 TI - [Hospital admission, duration of stay and mortality in community-acquired pneumonia in an acute care hospital. Correlation between a pneumonia prognosis index and conventional clinical criteria for assessing severity]. AB - INTRODUCTION: The objective of this study was to evaluate the management of community acquired pneumonia (CAP) according to conventional clinical criteria applied in the emergency room as compared to a pneumonia prognosis index (PPI) (Fine et al. NEJM 1997). We also analyzed which factors were associated with the need for inpatient treatment in PPI risk category III patients. METHODS: We prospectively enrolled all adults with CAP seen in the emergency room during 1999. The data required to calculate the PPI were collected at admission. Mortality and length of stay were recorded at discharge. RESULTS: A total of 447 patients with CAP were collected, 55.7% in the high-risk classes IV and V. Twenty seven patients died (6.1%) and 97% of these were within the high-risk classes. There were 362 hospitalizations; 302 (83%) were classes III, IV and V. The readmission rate increased with increases in the risk class, with a range of 4% for class I to 18% for class IV. Eighty-five patients (19%) were treated on an outpatient basis. Risk class III included 80 patients; 63 (79%) were hospitalized, with a length of stay of 7.89 days. The factor most highly associated with hospitalization in this group was abnormal findings on physical examination or on laboratory testing and radiographic studies. (OR: 7.62 [1.5 35.2]). CONCLUSION: In our cohort, the PPI was effective for identifying low-risk patients with CAP who could be treated as outpatients. In risk class III patients, the severity of the disease was the strongest predictor of hospitalization, rather than the presence of comorbid conditions. PMID- 14756987 TI - [Trichinellosis outbreaks in Spain (1990-2001)]. AB - INTRODUCTION: Trichinellosis is a helminth infection having a wide geographical distribution. It is acquired by ingestion of raw or undercooked meat infected with Trichinella spp. The present work reviews the human trichinellosis outbreaks recorded in Spain during 1990-2001, taking into account both the geographical distribution and source of the infection. METHODS: The parasite was isolated from suspected meats with the digestion technique. Molecular characterization of Trichinella isolates was done by random amplified polymorphic DNA (RAPD) analysis and Western-blot with US5 and US9 monoclonal antibodies. Anti-Trichinella antibodies in the patients' sera were investigated by indirect immunofluorescence (IIF). RESULTS: A total of 49 trichinellosis outbreaks were detected in Spain during 1990-2001, 75.5% produced by ingestion of infected wild boar, 14.3% by infected domestic pig and 10.2% by an unknown source. Of the 49 outbreaks, 21 were followed up in our laboratory. We were able to perform molecular identification of the Trichinella species involved in 13 outbreaks, with 61.5% produced by T. britovi and 38.5% by T. spiralis. Serological diagnosis of the trichinellosis patients by IIF allowed confirmation of helminth infection. CONCLUSIONS: With the use of molecular markers, T. spiralis and T. britovi were identified as the causative organisms in trichinellosis outbreaks detected in Spain during 1990-2001. These results reveal the importance of T. britovi in Spanish public health. PMID- 14756988 TI - [Poliomyelitis eradication initiative. Search for and control of wild poliovirus stored at Spanish laboratories]. AB - INTRODUCTION: An initiative to achieve global eradication of poliomyelitis was proposed in 1988. Since then, World Health Organization (WHO) efforts have been focused on reaching and maintaining high vaccination coverage and on implementing effective surveillance systems. Additionally, since 1998 the WHO has urged health authorities from countries over the world to locate and control the wild polioviruses stored in laboratories as well as potentially infectious materials that could contain these viruses or originate them under particular circumstances. METHODS: In Spain the Wild Poliovirus Containment Plan was initiated in 1999. All Spanish laboratories were inventoried and a specially designed questionnaire was distributed to them. The responses were analyzed. RESULTS: A total of 3209 laboratories were surveyed and 10 of them were found to contain infectious or potentially infectious material with wild poliovirus. The heads of these laboratories were informed about the Containment Plan and were requested to destroy this material under proper conditions. The laboratories that did not destroy the viruses adopted the BSL2/polio bio-safety measures. CONCLUSION: Despite its complexity, the search process was completed within the schedule established by the WHO, thanks to the efforts of health authorities from the Autonomous Communities. Spain reported its results to the WHO Regional Commission for the Certification of Poliomyelitis Eradication in May 2002. PMID- 14756989 TI - [Evaluation of a new commercial test (Candida albicans IFA IgG) for the serodiagnosis of invasive candidiasis]. AB - INTRODUCTION: Two tests for the detection of antibodies to Candida albicans germ tubes in patients with invasive candidiasis were compared: a new commercially available test (Candida albicans IFA IgG) and the indirect immunofluorescence test generally used for this purpose. METHODS: With the use of two indirect immunofluorescence tests, retrospective study was done on 172 sera from 51 patients classified into two groups: Group I included 123 serum samples from 32 patients with invasive candidiasis, and Group II, the control, included 49 serum samples from 19 patients with no evidence of Candida infection. RESULTS: In Group I, 84% of patients presented anti-germ tube antibody titers >or= 1:160 by the Candida albicans IFA IgG test and 78.1% of patients were positive by the generally used test. There was a high correlation between the two tests (R2 =0.9512 by patients; R2 = 0.8986 by sera). When a titer value of >or= 1:160 was used as cutoff, the Candida albicans IFA IgG test showed a sensitivity of 84.4% and a specificity of 94.7%, whereas the traditional test showed a sensitivity of 78.1% and a specificity of 100%. CONCLUSIONS: The commercially available Candida albicans IFA IgG test is similar to the test generally used for the detection of antibodies to C. albicans germ tubes and provides faster and easier diagnosis of invasive candidiasis in the clinical microbiology laboratory. PMID- 14756990 TI - [Clinical significance of Streptococcus agalactiae isolation from urine samples of outpatients from health care centers]. AB - INTRODUCTION: Streptococcus agalactiae is a well-recognized pathogen in the obstetric population and is a cause of invasive infection in adults with underlying diseases. Nevertheless, the role of S. agalactiae in urinary tract infections in the adult non-pregnant population treated in outpatient health care centers has been less extensively studied. METHODS: The clinical significance of S. agalactiae isolation in urine samples from health care center patients was studied in five areas of Spain over a period of six months. The study protocol included the collection of personal, epidemiological and clinical data. A total of 85 patients were studied; 79 were women (17 pregnant) and 6 men. RESULTS: Pyuria was found in 72.9% of the cases. A total of 26.4% (n = 18) of non-pregnant women and all the men had some type of chronic underlying disease and 35.3% (n = 24) had urinary disease. Among the pregnant women, 53% had asymptomatic bacteriuria and 35.3% had vaginal and/or urethral colonization. Among the group of men and non-pregnant women, the most frequent entity was non-complicated cystitis, seen in 66.1% (45 of 68) of cases, followed by complicated infection in 19.1% and asymptomatic bacteriuria in 11.8%. Of the 45 patients with non complicated cystitis, 35 (77.7%) were over 40 years old and 19 (42.2%) were over 60 years old. CONCLUSIONS: Among the population of non-pregnant adults treated in outpatient health care centers, S. agalactiae isolation from urine was found mainly in women over 40 years old and was the cause of non-complicated urinary tract infection in more than half of cases. PMID- 14756991 TI - [Guidelines for the treatment of short-term intravascular catheter-related infections in adults; SEIMC-SEMICYUC Consensus Conference]. PMID- 14756992 TI - [Importance of nosocomial transmission on severe acute respiratory syndrome and its prevention]. PMID- 14756993 TI - [Strategies to optimize adherence to antiretroviral treatment. Interventions in the therapeutic regimen]. AB - Within the first few years after the introduction of highly active antiretroviral therapy two facts became evident: the treatment was highly effective and proper compliance was essential to achieve the therapeutic objectives. Recently, the regimens containing these drugs have changed. Complex dosing with a large number of tablets taken three times daily together with dietary restrictions has given way to simpler treatments.The main advances include new formulations and new drugs that allow once-daily dosing, the use of ritonavir to enhance the bioavailability of the protease inhibitors and the coformulation of active ingredients in a single capsule. This review analyzes the impact of these interventions on the therapeutic regimen and it discusses the factors that facilitate and those that hinder optimal adherence to highly active antiviral treatment. Adherence is a complex, multidimensional problem. Simplification of the treatment is an important aspect, but it should be accompanied by other strategies focussed on the patient and the medical team in order to achieve effective long-term antiretroviral therapy in all patients. PMID- 14756994 TI - [Multilocus sequence typing: the molecular marker of the Internet era]. AB - Global or longer term epidemiology track the spread of clonal lineages, associated with hipervirulence or resistance or multi-resistance to antimicrobial agents. Therefore, the application of a molecular typing system for this purpose should produce data easily shared by different and geographically distant laboratories, as well as distinguish those clonal lineages even with low levels of variability accumulated in the genome.A marker based on the DNA sequence will produce objective results easily organized in data bases accessible by Internet. The application of a similar strategy that was used in the analysis of isoenzymes, by sequencing variable fragments of selected housekeeping genes, will allow obtaining a general view of the distribution of the clonal lineages and tracking their spread. PMID- 14756996 TI - [Carpal tunnel syndrome in a 68-year-old woman]. PMID- 14756995 TI - [Fever and left lower limb edema in a 39-year-old man]. PMID- 14756997 TI - [Urinary infection due to Haemophilus influenzae as initial manifestation of renal alterations]. PMID- 14756998 TI - [Submaxillary adenopathy due to Propionibacterium propionicum: atypical presentation]. PMID- 14756999 TI - [2004: Transcendental year for Enfermedades Infecciosas y Microbiologia Clinica]. PMID- 14757000 TI - [Publish or perish: perish for publish?]. PMID- 14757001 TI - [Hepatitis B virus surface antigen reactivity in the absence of antibodies to core antigen: an atypical serological pattern having diverse significance]. AB - BACKGROUND: Reactivity for hepatitis B virus (HBV) surface antigen (HBsAg) in samples lacking antibody to the HBV core antigen (anti-HBc) may be due to either non-specific reactions or sample contamination, but it can also reflect other situations that may present clinical relevance. In Spain, the frequency described for this atypical pattern of HBV markers ranged from 0.05 to 1.3% in different series, so that it can be considered as moderately frequent. METHODS: Confirmatory assays for HBsAg and tests for detecting total and IgM anti-HBc, HBV "e" antigen and viral DNA were performed on serum samples taken from 96 patients who showed reactivity for HBsAg in absence of total anti-HBc on routine studies. These samples were collected between January, 2001 and May, 2002 and were sent for study from different Spanish laboratories. Follow-up samples were also studied from selected cases. RESULTS: Presence of HBsAg was excluded in 70 cases (72.9%) and total anti-HBc was detected in two additional patients, so that the original results were confirmed in 24 patients (25.0%). After further investigations, two early phases of the window period of the acute HBV infection were identified, as well as a further case of chronic HBV carriage in absence of antibody response. A single case could be confirmed as due to a contamination of the aliquot of sample studied. The pattern of HBV markers and its evolution on the follow-up were the characteristic of the phenomenon known as "hepatitis B virus type 2 infection" in eight patients. No conclusions could be drawn from the remainder 12 cases, since no follow-up samples were available for study. CONCLUSIONS: Besides most cases respond to non-specific reactions or reflect situations of low clinical relevance, the reactivity for HBsAg in samples lacking anti-HBc should be taken into consideration and routinely investigated, since this atypical pattern can also reflect unusual, but clinically relevant facts of the HBV infection that must be noticed. PMID- 14757002 TI - [Microbiological, clinical and epidemiological aspects of Streptococcus pneumoniae isolates recovered over two years]. AB - INTRODUCTION: Streptococcus pneumoniae is the most frequent cause of non-hospital acquired pneumonia and meningitis in adults, and bacterial otitis media in children. Moreover, it causes a third of all acute sinusitis cases. Penicillin has been the treatment of choice for almost 50 years. Gradually, penicillin resistant pneumococci have appeared throughout the world. Our aim was to investigate the epidemiology, pattern of resistance and serotypes of Streptococcus pneumoniae infection in our area. METHODS: Over a period of two years (May 1997-May 1999), Streptococcus pneumoniae strains were isolated in the Clinical Microbiology Unit of the University Hospital Virgen de la Victoria in Malaga, Spain. This is a 750-bed hospital covering a population of 407,480 inhabitants, and admitting 21,500 hospitalized patients per year. Streptococcus was identified by standard procedures: serotyping was done with the Quellung test and antibiotic susceptibility study by the disk diffusion method and E-test. RESULTS: Streptococcus pneumoniae infection was diagnosed in 170 patients during the years studied. The microorganism was isolated from samples of sputum (82), blood (43), aspirated bronchial fluid, cerebrospinal fluid (6), and exudates (7). Non-hospital origin was identified in 88% of cases. The mean hospital stay was 12 days and mortality was 12.4%. Some 45.9% of the isolated strains were resistant to penicillin and 20% to cefotaxime. We found 31 different serotypes, with 77% of the isolated strains belonging to 12 serotypes. Serotypes 19, 3 and 6B were the most frequent in non-hospital infection, whereas 9V and 23F were related with nosocomial infection. Penicillin-resistant strains of Streptococcus pneumoniae belonged to 19 different serotypes; 6B, 9V, 14, 19 and 23F were the most important. CONCLUSIONS: As was expected, Streptococcus pneumoniae infections of mainly non-hospital origin in our area were characterized by elevated mortality and high-level resistance to penicillin. Immunosuppression was a predisposing factor. PMID- 14757004 TI - [Scientific production in infectious diseases in Spain (1991-2001): position within the European Union]. AB - INTRODUCTION: Medical research in our geographic area has undergone significant changes over the last three decades. The objective of this study was to quantitatively determine Spain's medical research output published in the journals included in the Infectious Diseases section of the Journal Citation Reports, and to compare it with that of other European Union (EU) countries. METHODS: The PubMed Web site (MEDLINE) was used to retrieve medical articles published by authors from Spain and other EU countries from 1991 to 2001 in 36 infectious disease journals included in the Infectious Diseases section of the Journal Citation Reports. RESULTS: We retrieved a total of 17,899 documents published by EU authors during the study period. Authors from Spanish institutions contributed 1,340 documents, 7.5% of the overall EU production. Spain ranked in the sixth position of the EU by number of papers. It remained in the same position after adjusting for gross national product and dropped to the ninth position after correcting for population. The number of articles published increased from 72 in the biennium 1991-1992 to 442 in the biennium 2000-2001. Spanish production rose by 514%, as compared with a median increase of 149% in the other EU countries. Spain was the first producer of articles from the EU in four journals: Diagnostic Microbiology and Infectious Disease (27.6%), European Journal of Clinical Microbiology & Infectious Diseases (24.8%), American Journal of Infection Control (21.8%) and Microbial Drug Resistance (17.5%). CONCLUSIONS: Spanish scientific production in infectious diseases has increased substantially in the period 1991-2001. PMID- 14757003 TI - [Listeriosis in patients with human immunodeficiency virus infection in Spain. Three new cases and literature review]. AB - INTRODUCTION: Listeriosis has been considered infrequent among patients infected with the human immunodeficiency virus (HIV). Only a limited number of listeriosis cases among HIV-infected patients have been published in Spain, where the prevalence of HIV infection is relatively high. We present our experience in this field and provide a review of the reported cases in our country. METHODS: We report three cases of listeriosis among the 903 HIV-infected patients attended in our service. Age, sex, stage of HIV infection, clinical form of listeriosis, treatment, and prognosis are analyzed in these three patients and in the published cases. RESULTS: Sixteen (76%) of the 21 cases analyzed were males. Ten had central nervous system infection, ten had bacteremia alone or bacteremia associated with another condition, and one had pneumonia. Two patients died (9.5%) and none of them were treated with the antibiotics of choice for listeriosis. CONCLUSION: Although Listeria monocytogenes infection is infrequent in HIV-infected patients (0.33% of patients attended in our service), clinicians should be alert to this possibility, since even the severe clinical forms have a favorable prognosis with proper treatment. PMID- 14757005 TI - [Primary HIV drug resistance in a prison population. REPRICOVA-2 Study]. AB - INTRODUCTION: Currently, there are few reports on primary human immunodeficiency virus (HIV) drug resistance in the prison population. METHODS: This is a descriptive, one-day prevalence study to identify HIV drug-resistant mutations in chronically infected treatment-naive prisoners. Systematic randomized sampling was performed and genotyping was done by automatic sequencing. RESULTS: A total of 90 patients were studied. Two samples were found to have nucleoside reverse transcriptase inhibitor (NRTI)-resistant mutations, four had non-nucleoside reverse transcriptase inhibitor (NNRTI)-resistant mutations and one had protease inhibitor (PI)-resistant mutations. CONCLUSIONS: There was a low rate of primary resistance in our series. Therefore, resistance testing is not required before prescribing initial antiretroviral therapy in these patients. PMID- 14757006 TI - [Guidelines for fungal diagnoses and antifungal sensitivity studies]. AB - The guidelines presented herein, which are based on the indications established by various studies and expert opinions, analyze several issues related to laboratory diagnosis of invasive fungal infections in immunosuppressed patients. PMID- 14757007 TI - [Cardiovascular risk in patients with chronic HIV-1 infection: a controversy with therapeutic, clinical and prognostic implications]. AB - Atherosclerosis increases cardiovascular risk and the possibility of developing acute myocardial infarction (AMI) or stroke. Patients infected with human immunodeficiency virus (HIV) often present morphological and metabolic alterations (hypercholesterolemia, hypertriglyceridemia, insulin resistance, diabetes) that can increase vascular risk. The frequent coexistence of classic risk factors (atherogenic diet, smoking, physical inactivity, cocaine abuse), the progressive increase in mean age of HIV-1 infected patients, and the polymedication they receive make it difficult to estimate the direct effect that new therapies may have on cardiovascular risk. Retrospective clinical studies with diverse designs in large cohorts offer contradictory results for cardiovascular risk in the HIV-infected population. Longer observational periods are needed and the effect of other classic risk factors needs to be controlled, in order to establish the possible detrimental effect the new therapies may have on cardiovascular risk in this population. PMID- 14757008 TI - [Basis, types and application of DNA arrays in clinical microbiology]. AB - The DNA microarrays or microchips are sets of DNA probes bound to a solid support in a prefixed and regular disposition. The target nucleic acid that can be detected is either DNA or RNA, which is previously labeled with a fluorochrome or a radioactive compound. The main advantage with respect to other molecular biological tools, such as polymerase chain reaction, is that thousands of genes can be detected in a single procedure. The application of the DNA arrays in the field of clinical microbiology is so far scarce. Among the specific applications we can point out: 1. Investigation of bacterial pathogenesis; 2. Analysis of bacterial evolution and molecular epidemiology; 3. Study of the mechanisms of action and resistance to antimicrobial agents and 4. Microbiological diagnostic of the infectious diseases. This methodology is still in an embryonic phase with respect to its application in clinical microbiology. However, it presents a series of advantages that make it very attractive and in the future it may become a valuable tool for the diagnosis of infectious diseases. PMID- 14757009 TI - [Acute endophthalmitis after cataract extraction]. PMID- 14757010 TI - [Febrile condition with decreased consciousness]. PMID- 14757011 TI - [Cholestatic hepatitis due to amoxicillin-clavulanic acid with positive re exposure: Importance of proper terminology in drug vigilance]. PMID- 14757012 TI - [Delay in the diagnosis and treatment of hospitalized patients with tuberculosis]. PMID- 14757013 TI - [Chronic lung disease/bronchopulmonary dysplasia in infants: what is the treatment?]. PMID- 14757014 TI - [Respiratory morbidity after hospital discharge in premature infants born at < or = 32 weeks gestation with bronchopulmonary dysplasia]. AB - BACKGROUND: Bronchopulmonary dysplasia (BPD) is the most frequent cause of respiratory morbidity in the first 2 years of life among preterm infants who survive the first 28 days. OBJECTIVES: To evaluate respiratory morbidity in the first 2 years of life in a group of preterm infants born at (32 weeks' gestation with BPD (oxygen requirement at 36 weeks' postconceptional age) by comparing it with that in preterm infants born at (32 weeks without BPD and with a control group of full term infants without neonatal morbidity. To determine whether respiratory morbidity in children with BPD decreases after the age of 2 years. PATIENTS AND METHOD: Group I: preterm children with BPD (n = 29). Group II: preterm children without BPD (n = 29). Group III: children with appropriate gestational age and weight (n = 32). A cross-sectional, descriptive study of the three groups was performed over a 2-year period. In 17 children in group 1, the study was prolonged to the age of 4 years. We analyzed wheezing on at least two occasions, use of inhaled bronchodilators, use of inhaled glucocorticosteroids for more than 6 months, and hospitalization for respiratory illness. The chi square test and Fischer's exact test were performed. RESULTS: At least one episode of wheezing occurred in 25 children (86.2%) in group I compared with 12 children (41.4%) in group II and 6 (18.8%) in group III. Nineteen children (65.5%) in group I and none in the remaining two groups received treatment with inhaled glucocorticosteroids for more than 6 months (p < 0.001). Inhaled bronchodilators were used by 25 children (86.2%) in group I compared with 12 (41.4%) in group II and 6 (18.8%) in the control group (p < 0.001). Twelve children (41.3%) in group I were hospitalized for respiratory illness compared with 8 (27.6%) in group II. There were no admissions among the control group. None of the children with BPD who received prophylaxis with palivizumab contracted respiratory syncytial virus infection. Seventeen children with BPD were evaluated until the age of 4 years. Episodes of wheezing decreased from 88.2% in the first year to 41 % between the third and fourth years (p < 0.001). Treatment with inhaled glucocorticosteroids for more than 6 months was given to 88.2% in the first year, 41.2 % between the first and second year and to 0 % after the second year (p < 0.001). Hospital admissions for respiratory illness decreased from 52.9% in the first year to 17.6% in the second year. None of the children were hospitalized after the age of 2 years (p < 0.001). CONCLUSIONS: During the first 2 years of life, children with BPD showed a greater number of admissions and episodes of wheezing and a greater need for medical treatment. Respiratory morbidity improved with age, 40% showed recurrent wheezing episodes at the age of 4 years. PMID- 14757015 TI - [Epidemiology and burden of acute otitis media in Valencia (Spain)]. AB - OBJECTIVE: To assess the burden (incidence, treatment and complications) of acute otitis media (AOM) and otitis media with effusion (OME) in children younger than 5 years of age from Valencia, Spain. SUBJECT AND METHODS: We performed a retrospective cohort study of 1,399 children followed-up for the first 5 years of life. Seventeen pediatricians reviewed the medical records of their patients born in 1995 and 1996 and followed-up from birth until the age of 5 years. For each child, the number of otitis episodes, treatment, complications, and surgical interventions was obtained. RESULTS: There were 2,961 episodes of AOM in the first 5 years of life (2.23 cases/child). Four hundred seventy-six cases (16.1 %) occurred before 1 year of age and 1,346 between the first and second year of life (45.5 %). By the third year of life, 59.8 % had had at least one episode. In most children (80.9 %), diagnosis was made in primary care and required a median of 1.81 visits/episode for follow-up. A total of 94.5 % were treated with antibiotics (amoxicillin-clavulanate 38.8 %, cefuroxime 14.3 %, clarithromycin 8.2 % and amoxicillin 5.9 %) and 8.5 % required a change of antibiotic therapy. Two hundred seventeen children (15.2 %) had at least one episode of OME. Twenty six patients (1.8 %, 95 % CI: 1.2-2.7 %) required insertion of ventilation tubes. Twenty-four patients (1.7 %) had secondary hypoacusis. There was one case of meningitis and two cases of chronic otorrhea. No cases of mastoiditis were recorded. CONCLUSIONS: The incidence of AOM in Valencia is 40,014 episodes/100,000 children younger than 5 years/year (95 % CI: 39,700-40,300). It represents a significant burden due to the large number of visits, antibiotic use, associated surgical procedures and need for auditory rehabilitation. PMID- 14757016 TI - [Incidence, air pollution and risk factors of acute otitis media in the first year of life: a prospective study]. AB - BACKGROUND: Epidemiological studies of acute otitis media (AOM) are scarce and no prospective studies have been performed in Spain. OBJECTIVES: To describe the incidence of AOM in the first year of life and its associated risk factors, with special focus on air pollution. METHODS: We performed a prospective cohort study of 229 newborn infants during the first year of life stratified by pollution zones, and followed-up by their pediatricians in their health center. AOM was defined on clinical grounds. A questionnaire on risk/protective factors included items on the following: sex, older siblings, smoking, breastfeeding, socioeconomic status, parental education and the mother's occupational status. RESULTS: The incidence of AOM episodes during the first year of life was 45 % and the proportion of children who experienced at least one episode was 32 %. Independent risk factors were male gender (aOR: 2.03; 95 % CI: 1.09-3.7) and living in a polluted area (aOR: 2.01; 95 % CI: 1.05-3.84). Independent protective factors were being born in spring (aOR: 0.41; 95 % CI: 0.19-0.88) and having a mother with at least primary school education (aOR: 0,53; 95 % CI: 0.24-1.15). Socioeconomic markers indicated a lower mean level among families whose children had at least one AOM episode. CONCLUSIONS: Air pollution and low socioeconomic status are greater risk factors for AOM than having siblings or parents who smoke. A minimum educational level reduces the risk of AOM. The incidence of AOM could be reduced by modifying certain environmental factors. PMID- 14757017 TI - [Exposure to smoking during pregnancy: Barcelona (Spain) 1994-2001]. AB - BACKGROUND: Exposure to smoking during pregnancy is an important risk factor for child health. In the last few decades, the prevalence of smoking among fertile women has increased in Spain. OBJECTIVES: To assess fetal exposure to smoking in a representative sample of pregnant women and its trends. METHODS: The prevalence of smoking and smoking cessation were analyzed in a representative sample of women who gave birth in the city of Barcelona, extracted from a population-based registry of birth defects. The sample consisted of 1,801 women and covered the period 1994 to 2001. RESULTS: Among pregnant women, 43.4 % smoked before pregnancy, and 42.2 % of these quit. At the time of giving birth, 25.2 % of the women were smokers. Over the period studied the prevalence of smoking among pregnant women clearly decreased. Although the proportion of cessations showed no significant changes, fetal exposure to tobacco decreased. CONCLUSIONS: The prevalence of smoking among pregnant women was high, although many quit during pregnancy. Over the period studied, fetal exposure to smoking decreased. There is a need for more systematic interventions. PMID- 14757018 TI - [Metabolic alterations related to syndrome X and low vitamin E levels in obese children with acanthosis nigricans]. AB - BACKGROUND: In adults, both metabolic alterations related to syndrome X and lower plasma vitamin E levels have been associated with an increased risk of developing cardiovascular disease. OBJECTIVES: To study the presence of metabolic alterations related to syndrome X and to determine the plasma levels of vitamin E in obese children with acanthosis nigricans. METHODS: We performed a prospective study in 42 obese children [15 with acanthosis nigricans (AN) and 27 without]. Thirteen healthy non-obese children were also studied. After a 12-hour fast, plasma levels of glucose, insulin, triglycerides, cholesterol, high-density lipoprotein (HDL)-cholesterol, and vitamin E were determined. The insulin resistance index was also calculated. Differences between groups were determined using ANOVA. RESULTS: Obese children with AN showed higher plasma levels of insulin and triglycerides and lower plasma levels of HDL-cholesterol and vitamin E, as well as a higher insulin resistance index than non-obese children and obese children without AN. CONCLUSIONS: In obese children, the presence of acanthosis nigricans is linked to a group of metabolic alterations associated with a higher risk of developing cardiovascular disease and type 2 diabetes. PMID- 14757019 TI - [Follow-up of longstanding ventricular septal defects]. AB - OBJECTIVE: Ventricular septal defect is the most frequently diagnosed congenital heart defect. The prognosis is usually good. The aim of this study was to describe this idea to general pediatricians. MATERIAL AND METHODS: We review the follow-up of 81 patients with ventricular septal defect. Defects that spontaneously closed in the first 12 months of life and those that formed part of a malformative syndrome or a complex congenital heart defect were excluded. RESULTS: Localization was perimembranous, including all defects affecting mainly the septal membrane independent of whether the surrounding tissues were involved, in 66.7 %, muscular in 29.6 % and mixed in 3.7 %. Perimembranous position was more frequent among large and medium-sized defects. Large and perimembranous defects were characterized by holosystolic murmur; in small, muscular defects, murmur was cut off in mid-systole. In 45.8 % of large defects, weight development was delayed, but there was no appreciable effect on height. Generally we observed a tendency to partial closure and to improvement. Surgical closure was required in 9.8 %. CONCLUSIONS: Because of the trend to partial or complete spontaneous closure, the prognosis of ventricular septal defect is generally good. PMID- 14757020 TI - [Magnetic resonance imaging and lung perfusion scintigraphy in tetralogy of Fallot following surgery]. AB - OBJECTIVE: To evaluate pulmonary arteries in patients with tetralogy of Fallot following surgery with quantified lung perfusion scintigraphy and magnetic resonance imaging. MATERIAL AND METHODS: From January 1985 to December 1999, 47 patients who underwent surgery between 1985 and 1999 were studied. To obtain values of normality, 45 infants with no pulmonary artery disease were assessed with lung perfusion scintigraphy (right lung flow: 54-61%, left lung flow: 38.7 46%) and magnetic resonance imaging axial view (right/left pulmonary artery branch diameter ratio: 1-1.1). Patients with stenosis underwent catheterization. RESULTS: Group 1: 27 patients with normal parameters. Group 2: nine patients with left pulmonary branch stenosis and irregularities in all parameters; left/right branch diameter ratio 0.51 and left lung perfusion 26.3 +/- 7.9%, r correlation ratio 65%, p < 0.005, left branch pressure gradient 34.4 +/- 17.9 mm Hg, rate gradient r 89%, p < 0.001. Group 3: five patients with right pulmonary branch stenosis and irregularities in all parameters; right/left branch diameter ratio 0.52, reduced right lung perfusion 32 +/- 11%, with a stenotic branch gradient of 40 +/- 19 mm Hg., rate/gradient ratio r 72%, p < 0.005; gradient/perfusion ratio r 82%, p < 0.003. Group 4: Six patients with bilateral stenosis, reduced diameter in the stenotic area with normal perfusion in both lungs and bilateral gradient. Eighteen patients with stenosis underwent re-operation, with favorable outcome in 14. CONCLUSION: Magnetic resonance imaging and quantified lung perfusion scintigraphy provide useful information in the follow-up of tetralogy of Fallot. PMID- 14757021 TI - [Drinking water in infants]. AB - We review types of public drinking water and bottled water and provide recommendations on the composition of water for infants. Water used with any of the commercial infant formulas in Spain should contain less than 25 mg/l of sodium. Drinking water must be boiled for a maximum of one minute (at sea level) to avoid excessive salt concentration. Bottled water need not be boiled. Fluoride content in drinking water should be less than 0.3 mg/l in first year of life to prevent dental fluorosis. Nitrate content in water should be less than 25 mg/l to prevent methemoglobinemia. Water with a calcium concentration of between 50 and 100 mg/l is a dietary source of calcium since it provides 24-56 % of the required daily intake in infancy. PMID- 14757022 TI - [Biliary lithiasis in childhood: therapeutic approaches]. AB - Until recently, biliary lithiasis was considered infrequent in childhood. According to their composition, gallstones can be classified into cholesterol stones and pigment stones. The latter are mainly composed of calcium salts of unconjugated bilirubin and are divided into hard black and soft brown stones. In children, up to 75 % of gallstones are pigment stones. Their etiology is often unknown. Biliary lithiasis in children differs from that in adults and there is very little scientific evidence on the most suitable therapeutic procedures. Symptom-free stones usually have a benign course and do not require medical or surgical treatment. Symptoms are often nonspecific and include dyspepsia and chronic abdominal pain. These symptoms are an indication for ultrasonographic scan to rule out the presence of gallstones. Cholecystectomy is the definitive treatment for gallstones but is not always indicated. Medical treatment with ursodeoxycholic acid is indicated in oligosymptomatic and asymptomatic lithiasis with transparent, soft, cholesterol-rich stones and a functional bladder and in patients with a high surgical risk. PMID- 14757023 TI - [Persistent dysphagia in a 9-year-old girl]. PMID- 14757024 TI - [Paracetamol poisoning in infants aged less than six months: dosage errors]. AB - In infants under 6 months of age, paracetamol overdose is usually due to dose confusion by caretakers. Recently, liquid formulations of this drug have been commercialized in larger,60-ml bottles. The syringe to measure the syrup in these new formulations is also bigger (5 cc versus 1.2-2 cc). We present six cases of 2 4-month-old infants mistakenly given an overdose of paracetamol, each from this new 60-ml formulation. These patients are especially susceptible to poisoning because of liver immaturity and require more aggressive management. To prevent this kind of poisoning, correct and clear information must be given to caregivers about drug dosage. PMID- 14757025 TI - [Tetralogy of Fallot with absent pulmonary valve in a newborn and infant. Complete surgical correction]. AB - We report two patients, a newborn and a 7-month old infant, with tetralogy of Fallot and absent pulmonary valve syndrome. Both had severe obstruction at the level of the ring with aneurysmal pulmonary artery branches, which compressed and displaced the trachea and main bronchial tubes. The neonate required mechanical ventilation from birth. Treatment was aggressive in both patients with interventricular septum defect closure, arterioplasty of the branches and homograft in the infant, and resection of the truncus and pulmonary branches with posterior face suture of both branches associated with a valved conduit in orthotopic position in the neonate. We believe that early treatment avoids airway degeneration and right ventricle volume overload. PMID- 14757026 TI - [Listeria monocytogenes meningitis in an immunocompetent girl]. PMID- 14757027 TI - [Lumbar disc herniation in a 10-year old girl]. PMID- 14757028 TI - [Acute non-traumatic paraparesis]. PMID- 14757029 TI - [Cerebral vasculitis in Henoch-Schonlein purpura]. PMID- 14757030 TI - [Change in attitude to bicycle helmets after serious accidents]. PMID- 14757031 TI - [Thoracic ectopia cordis]. PMID- 14757032 TI - [Scabies in infant]. PMID- 14757033 TI - [Prevention of adverse effects related to ophthalmological examination in premature infants]. PMID- 14757034 TI - Where, oh where has my protein gone? AB - A recent publication by R. Chikwamba and colleagues highlights interesting issues in recombinant protein expression in transgenic plants. In the study they expressed a bacterial antigen in maize seed and obtained aberrant localization data. This work is of great importance to the biotechnology industry and raises fascinating questions in plant cell biology that require creative thinking. PMID- 14757035 TI - Surface-attached molecular beacons light the way for DNA sequencing. AB - Rapid testing of DNA and RNA nucleotide sequences is required for various research protocols including wide-scale genetic testing, diagnostics, fast detection of biological warfare agents, environmental testing and forensic medicine. At present many laboratories are interested in research and development of an inexpensive, easy-to-use, fast-response device for this purpose. Various methods based on acoustic, electronic and optical detection of the DNA hybridization event have been reported. PMID- 14757036 TI - Tracking the 'general': tagging skin-derived dendritic cells. AB - Dendritic cells (DCs) are the sentinels of the immune system; their migration, maturation and mobilization are fundamental to immunity and tolerance. The recent tracking of DCs from the skin to lymph node (LN) and their enumeration using a Cre/loxP system demonstrate the recruitment of a higher than expected number of DCs to the draining LN after cutaneous administration of DNA-coated gold particles. The longevity of the migrated DCs was also longer than previously reported. PMID- 14757037 TI - Enhanced functional information from predicted protein networks. AB - Experimentally derived genome-wide protein interaction networks have been useful in the elucidation of functional information that is not evident from examining individual proteins but determination of these networks is complex and time consuming. To address this problem, several computational methods for predicting protein networks in novel genomes have been developed. A recent publication by Date and Marcotte describes the use of phylogenetic profiling for elucidating novel pathways in proteomes that have not been experimentally characterized. This method, in combination with other computational methods for generating protein interaction networks, might help identify novel functional pathways and enhance functional annotation of individual proteins. PMID- 14757039 TI - On risk and plant-based biopharmaceuticals. AB - Research into plant-based expression of pharmaceutical proteins is proceeding at a blistering pace. Indeed, plants expressing pharmaceutical proteins are currently being grown in field environments throughout the USA. But how are these plants and proteins being assessed for environmental risk and how are they being regulated? Here, we examine the applicability of the risk assessment paradigm for assessing human and ecological risks from field-grown transgenic plants that express pharmaceutical proteins. PMID- 14757040 TI - Confessions of a bioenergy advocate. AB - Feedstocks that deserve serious consideration for fuels and chemicals are sugarcane, corn, trees and algae. Commercialization of biomass refining is imminent but the wild claims of those who think that bioenergy can replace much of our dependence on foreign oil are appalling. It is naive to view biomass as the panacea for the coming energy crisis because there is not enough in practical locations and the costs involved in retrieving and refining it will be relatively high. The world will not run out of energy, but cheap energy might disappear, with its economics clouded by a myriad of subsidies for the competing energy sources and by world politics. This assessment of biomass supply and conversion technologies provides global perspectives and exposes some alternatives to be so impractical that they are almost fraudulent. PMID- 14757041 TI - Engineering metabolic highways in Lactococci and other lactic acid bacteria. AB - Lactic acid bacteria (LAB) are widely used in industrial food fermentations and are receiving increased attention for use as cell factories for the production of food and pharmaceutical products. Glycolytic conversion of sugars into lactic acid is the main metabolic highway in these Gram-positive bacteria and Lactococcus lactis has become the model organism because of its small genome, genetic accessibility and simple metabolism. Here we discuss the metabolic engineering of L. lactis and the value of metabolic models compared with other LAB, with a particular focus on the food-grade production of metabolites involved in flavour, texture and health. PMID- 14757042 TI - The role of bioreactors in tissue engineering. AB - Ex vivo engineering of living tissues is a rapidly developing area with the potential to impact significantly on a wide-range of biomedical applications. Major obstacles to the generation of functional tissues and their widespread clinical use are related to a limited understanding of the regulatory role of specific physicochemical culture parameters on tissue development, and the high manufacturing costs of the few commercially available engineered tissue products. By enabling reproducible and controlled changes of specific environmental factors, bioreactor systems provide both the technological means to reveal fundamental mechanisms of cell function in a 3D environment, and the potential to improve the quality of engineered tissues. In addition, by automating and standardizing tissue manufacture in controlled closed systems, bioreactors could reduce production costs, thus facilitating a wider use of engineered tissues. PMID- 14757043 TI - History, challenges and perspectives of cell microencapsulation. AB - Cell microencapsulation continues to hold significant promise for biotechnology and medicine. The controlled, and continuous, delivery of therapeutic products to the host by immunoisolated cells is a potentially cost-effective method to treat a wide range of diseases. Although there are several issues that need to be addressed, including capsule manufacture, properties and performance, in the past few years, a stepwise analysis on the essential obstacles and limitations has brought the whole technology closer to a realistic proposal for clinical application. This paper summarizes the current situation in the cell encapsulation field and discusses the main events that have occurred along the way. PMID- 14757044 TI - Engineering amyloidogenicity towards the development of nanofibrillar materials. AB - When folded into their native structures, proteins in biological systems function as nanostructured machines. By contrast, some polypeptides tend to aggregate into other well-ordered structures, namely amyloid fibrils. Such well-ordered protein fibrils are attractive materials for nanobiotechnology because they self associate through noncovalent bonds under controlled conditions - a property that is shared with small organic molecules called organogelators. Recently, the use of amyloid fibrils as structural templates for constructing nanowires has been demonstrated. Such applications will potentially become one of the next trends in protein engineering and nanobiotechnology. PMID- 14757045 TI - Target specificity analysis of the Abl kinase using peptide microarray data. AB - Protein kinases play an important role in cellular signalling. The reliable prediction of their substrates is of high importance for the deciphering of signalling pathways. A recently developed peptide microarray technology for the charcterisation of protein kinases delivers data on the individual phosphorylation status of each single member of a large peptide library. This data can be used to approximate the substrate specificity of the investigated kinase. We present an approach to process the collected information using a combination of a weight matrix approach and a nearest neighbor approach. Experiments with the protein-tyrosine kinase Abl are conducted to validate the results. Randomly selected peptides (1433) are used to estimate the substrate preferences of the kinase. The obtained prediction results are compared with standard methods. The new approach is tested further on bona fide Abl phosphorylation sites. PMID- 14757046 TI - Relation between protein stability, evolution and structure, as probed by carboxylic acid mutations. AB - Native proteins are marginally stable. Low thermodynamic stability may actually be advantageous, although the accumulation of neutral, destabilizing mutations may have also contributed to it. In any case, once marginal stability has been reached, it appears plausible that mutations at non-constrained positions become fixed in the course of evolution (due to random drift) with frequencies that roughly reflect the mutation effects on stability ("pseudo-equilibrium hypothesis"). We have found that all glutamate-->aspartate mutations in wild-type Escherichia coli thioredoxin are destabilizing, as well as most of the aspartate- >glutamate mutations. Furthermore, the effect of these mutations on thioredoxin thermodynamic stability shows a robust correlation with the frequencies of occurrence of the involved residues in several-hundred sequence alignments derived from a BLAST search. These results provide direct and quantitative experimental evidence for the pseudo-equilibrium hypothesis and should have general consequences for the interpretation of mutation effects on protein stability, as they suggest that residue environments in proteins may be optimized for stabilizing interactions to a remarkable degree of specificity. We also provide evidence that such stabilizing interactions may be detected in sequence alignments, and briefly discuss the implications of this possibility for the derivation of structural information (on native and denatured states) from comparative sequence analyses. PMID- 14757047 TI - Structure of the NCoA-1/SRC-1 PAS-B domain bound to the LXXLL motif of the STAT6 transactivation domain. AB - Signal transducer and activator of transcription 6 (STAT6) regulates transcriptional activation in response to interleukin-4 (IL-4) by direct interaction with coactivators. The CREB-binding protein (p300/CBP) and the nuclear coactivator 1 (NCoA-1), a member of the p160/steroid receptor coactivator family, bind independently to specific regions of the STAT6 transactivation domain and act as coactivators. The interaction between STAT6 and NCoA-1 is mediated by an LXXLL motif in the transactivation domain of STAT6. To define the mechanism of coactivator recognition, we determined the crystal structure of the NCoA-1 PAS-B domain in complex with the STAT6 LXXLL motif. The amphipathic, alpha helical STAT6 LXXLL motif binds mostly through specific hydrophobic interactions to NCoA-1. A single amino acid of the NCoA-1 PAS-B domain establishes hydrophilic interactions with the STAT6 peptide. STAT6 interacts only with the PAS-B domain of NCoA-1 but not with the homologous regions of NCoA-2 and NCoA-3. The residues involved in binding the STAT6 peptide are strongly conserved between the different NCoA family members. Therefore surface complementarity between the hydrophobic faces of the STAT6 fragment and of the NCoA-1 PAS-B domain almost exclusively defines the binding specificity between the two proteins. PMID- 14757048 TI - Functional compatibility of elongation factors between mammalian mitochondrial and bacterial ribosomes: characterization of GTPase activity and translation elongation by hybrid ribosomes bearing heterologous L7/12 proteins. AB - The mammalian mitochondrial (mt) ribosome (mitoribosome) is a bacterial-type ribosome but has a highly protein-rich composition. Almost half of the rRNA contained in the bacterial ribosome is replaced with proteins in the mitoribosome. Escherichia coli elongation factor G (EF-G Ec) has no translocase activity on the mitoribosome but EF-G mt is functional on the E.coli ribosome. To investigate the functional equivalency of the mt and E.coli ribosomes, we prepared hybrid mt and E.coli ribosomes. The hybrid mitoribosome containing E.coli L7/12 (L7/12 Ec) instead of L7/12 mt clearly activated the GTPase of EF-G Ec and efficiently promoted its translocase activity in an in vitro translation system. Thus, the mitoribosome is functionally equivalent to the E.coli ribosome despite their distinct compositions. The mt EF-Tu-dependent translation activity of the E.coli ribosome was also clearly enhanced by replacing the C-terminal domain (CTD) of L7/12 Ec with the mt counterpart (the hybrid E.coli ribosome). This strongly indicates that the CTD of L7/12 is responsible for EF-Tu function. These results demonstrate that functional compatibility between elongation factors and the L7/12 protein in the ribosome governs its translational specificity. PMID- 14757049 TI - Rational design of inhibitors of HIV-1 TAR RNA through the stabilisation of electrostatic "hot spots". AB - The targeting of RNA for the design of novel anti-viral compounds has until now proceeded largely without incorporating direct input from structure-based design methodology, partly because of lack of structural data, and complications arising from substrate flexibility. We propose a paradigm to explain the physical mechanism for ligand-induced refolding of trans-activation response element (TAR RNA) from human immunodeficiency virus 1 (HIV-1). Based upon Poisson-Boltzmann analysis of the TAR structure, as bound by a peptide derived from the transcriptional activator protein, Tat, our hypothesis shows that two specific electrostatic interactions are necessary to stabilise the conformation. This result contradicts the belief that a single argininamide residue is responsible for stabilising the TAR fold, as well as the conventional wisdom that electrostatic interactions with RNA are non-specific or dominated by phosphates. We test this hypothesis by using NMR and computational methods to model the interaction of a series of novel inhibitors of the in vitro RNA-binding activities for a peptide derived from Tat. A subset of inhibitors, including the bis-guanidine compound rbt203 and its analogues, induce a conformation in TAR similar to that brought about by the protein. Comparison of the interactions of two of these ligands with the RNA and structure-activity relationships observed within the compound series, confirm the importance of the two specific electrostatic interactions in the stabilisation of the Tat-bound RNA conformation. This work illustrates how the use of medicinal chemistry and structural analysis can provide a rational basis for prediction of ligand-induced conformational change, a necessary step towards the application of structure based methods in the design of novel RNA or protein-binding drugs. PMID- 14757050 TI - Molecular interplay between RNA polymerase and two transcriptional regulators in promoter switch. AB - Transcription regulation relies in the molecular interplay between the RNA polymerase (RNAP) and regulatory factors. Phage phi29 promoters A2c, A2b and A3 are coordinately regulated by the transcriptional regulator protein p4 and the histone-like protein p6. This study shows that protein p4 binds simultaneously to four sites: sites 1 and 2 located between promoters A2c and A2b and sites 3 and 4 between promoters A2b and A3, placed in such a way that bound p4 is equidistant from promoters A2c and A2b and one helix turn further upstream from promoter A3. The p4 molecules bound to sites 1 and 3 reorganise the binding of protein p6, giving rise to the nucleoprotein complex responsible for the switch from early to late transcription. We identify the positioning of the alphaCTD-RNAP domain at these promoters, and demonstrate that the domains are crucial for promoter A2b recognition and required for full activity of promoter A2c. Since binding of RNAP overlaps with p4 and p6 binding, repression of the early transcription relies on the synergy of the regulators able to antagonize the stable binding of the RNAP through competition for the same target, while activation of late transcription is carried out through the stabilization of the RNAP by the p4/p6 nucleoprotein complex. The control of promoters A2c and A2b by feed-back regulation is discussed. PMID- 14757051 TI - RNA interactions in the 5' region of the HIV-1 genome. AB - The untranslated leader of the dimeric HIV-1 RNA genome is folded into a complex structure that plays multiple and essential roles in the viral replication cycle. Here, we have investigated secondary and tertiary structural elements within the 5' 744 nucleotides of the HIV-1 genome using a combination of bioinformatics, enzymatic probing, native gel electrophoresis, and UV-crosslinking experiments. We used a recently developed RNA folding algorithm (Pfold) to predict the common secondary structure of an alignment of 20 divergent HIV-1 sequences. Combining this analysis with biochemical data, we present a secondary structure model for the entire 744 nucleotide fragment, which incorporates previously recognized and novel structural elements. In particular, our data provided strong evidence for a long-distance interaction between the region encompassing the AUG Gag initiation codon and an upstream region and we demonstrate that this feature is highly conserved in distantly related human and animal retroviruses. To obtain information about tertiary interactions we applied an intramolecular UV crosslinking strategy and identified a novel tertiary interaction within the PBS hairpin structure. PMID- 14757052 TI - The clamp-loader-helicase interaction in Bacillus. Atomic force microscopy reveals the structural organisation of the DnaB-tau complex in Bacillus. AB - The clamp-loader-helicase interaction is an important feature of the replisome. Although significant biochemical and structural work has been carried out on the clamp-loader-clamp-DNA polymerase alpha interactions in Escherichia coli, the clamp-loader-helicase interaction is poorly understood by comparison. The tau subunit of the clamp-loader mediates the interaction with DnaB. We have recently characterised this interaction in the Bacillus system and established a tau(5) DnaB(6) stoichiometry. Here, we have obtained atomic force microscopy images of the tau-DnaB complex that reveal the first structural insight into its architecture. We show that despite the reported absence of the shorter gamma version in Bacillus, tau has a domain organisation similar to its E.coli counterpart and possesses an equivalent C-terminal domain that interacts with DnaB. The interaction interface of DnaB is also localised in its C-terminal domain. The combined data contribute towards our understanding of the bacterial replisome. PMID- 14757053 TI - DNA-binding orientation and domain conformation of the E. coli rep helicase monomer bound to a partial duplex junction: single-molecule studies of fluorescently labeled enzymes. AB - The SF1 DNA helicases are multi-domain proteins that can unwind duplex DNA in reactions that are coupled to ATP binding and hydrolysis. Crystal structures of two such helicases, Escherichia coli Rep and Bacillus stearothermophilus PcrA, show that the 2B sub-domain of these proteins can be found in dramatically different orientations (closed versus open) with respect to the remainder of the protein, suggesting that the 2B domain is highly flexible. By systematically using fluorescence resonance energy transfer at the single-molecule level, we have determined both the orientation of an E.coli Rep monomer bound to a 3' single-stranded-double-stranded (ss/ds) DNA junction in solution, as well as the relative orientation of its 2B sub-domain. To accomplish this, we developed a highly efficient procedure for site-specific fluorescence labeling of Rep and a bio-friendly immobilization scheme, which preserves its activities. Both ensemble and single-molecule experiments were carried out, although the single-molecule experiments proved to be essential here in providing quantitative distance information that could not be obtained by steady-state ensemble measurements. Using distance-constrained triangulation procedures we demonstrate that in solution the 2B sub-domain of a Rep monomer is primarily in the "closed" conformation when bound to a 3'-ss/ds DNA, similar to the orientation observed in the complex of PcrA bound to a 3'-ss/ds DNA. Previous biochemical studies have shown that a Rep monomer bound to such a 3'-ss/ds DNA substrate is unable to unwind the DNA and that a Rep oligomer is required for helicase activity. Therefore, the closed form of Rep bound to a partial duplex DNA appears to be an inhibited form of the enzyme. PMID- 14757054 TI - Crystal structure of a gamma-butyrolactone autoregulator receptor protein in Streptomyces coelicolor A3(2). AB - The gamma-butyrolactone-type autoregulator/receptor systems in the Gram-positive bacterial genus Streptomyces regulate morphological differentiation or antibiotic production, or both. The autoregulator receptors act as DNA-binding proteins, and on binding their cognate ligands (gamma-butyrolactones) they are released from the DNA, thus serving as repressors. The crystal structure of CprB in Streptomyces coelicolor A3(2), a homologue of the A-factor-receptor protein, ArpA, in Streptomyces griseus, was determined. The overall structure of CprB shows that the gamma-butyrolactone receptors belong to the TetR family. CprB is composed of two domains, a DNA-binding domain and a regulatory domain. The regulatory domain contains a hydrophobic cavity, which probably serves as a ligand-binding pocket. On the basis of the crystal structure of CprB and on the analogy of the characteristics of ligand-TetR binding, the binding of gamma butyrolactones to the regulatory domain of the receptors is supposed to induce the relocation of the DNA-binding domain through conformational changes of residues located between the ligand-binding site and the DNA-binding domain, which would result in the dissociation of the receptors from their target DNA. PMID- 14757055 TI - Use of computer-designed group II introns to disrupt Escherichia coli DExH/D-box protein and DNA helicase genes. AB - Mobile group II introns are site-specific retroelements that use a novel mobility mechanism in which the excised intron RNA inserts directly into a DNA target site and is then reverse transcribed by the associated intron-encoded protein. Because the DNA target site is recognized primarily by base-pairing of the intron RNA with only a small number of positions recognized by the protein, it has been possible to develop group II introns into a new type of gene targeting vector ("targetron"), which can be reprogrammed to insert into desired DNA targets simply by modifying the intron RNA. Here, we used databases of retargeted Lactococcus lactis Ll.LtrB group II introns and a compilation of nucleotide frequencies at active target sites to develop an algorithm that predicts optimal Ll.LtrB intron-insertion sites and designs primers for modifying the intron to insert into those sites. In a test of the algorithm, we designed one or two targetrons to disrupt each of 28 Escherichia coli genes encoding DExH/D-box and DNA helicase-related proteins and tested for the desired disruptants by PCR screening of 100 colonies. In 21 cases, we obtained disruptions at frequencies of 1-80% without selection, and in six other cases, where disruptants were not identified in the initial PCR screen, we readily obtained specific disruptions by using the same targetrons with a retrotransposition-activated selectable marker. Only one DExH/D-box protein gene, secA, which was known to be essential, did not give viable disruptants. The apparent dispensability of DExH/D-box proteins in E.coli contrasts with the situation in yeast, where the majority of such proteins are essential. The methods developed here should permit the rapid and efficient disruption of any bacterial gene, the computational analysis provides new insight into group II intron target site recognition, and the set of E.coli DExH/D-box protein and DNA helicase disruptants should be useful for analyzing the function of these proteins. PMID- 14757056 TI - Conservation and variation of structure and function in a newly identified GCM homolog from chicken. AB - Glial cell missing (GCM) proteins constitute a small family of transcription factors with two members each described in Drosophila and several mammalian species. Here, we report the identification of a GCM homolog from chicken. Although the exon-intron structure is well conserved between chicken GCM and other family members, sequence similarity is largely restricted to the DNA binding GCM-domain (residues 24-176). In accord with the high degree of sequence conservation within the GCM-domain, the chicken GCM protein has a DNA-binding specificity similar to that of other GCM proteins. Like other GCM proteins, it is located to the nucleus and can act as a transcriptional activator despite the strong divergence in sequences outside the GCM-domain. The chicken GCM protein contains two transactivation domains with cell-specific function, one immediately following the DNA-binding domain, the other at its extreme carboxy terminus. Intriguingly, chicken GCM is expressed only transiently during embryogenesis and is restricted exclusively to extraembryonic tissues where it was detected in close vicinity to embryonic blood vessels. Taking the extraembryonic expression of chicken GCM and mammalian GCMa into account, it is tempting to speculate that a conserved extraembryonic function exists for GCM proteins in birds and mammals. PMID- 14757057 TI - Recognition and separation of single particles with size variation by statistical analysis of their images. AB - Macromolecules may occupy conformations with structural differences that cannot be resolved biochemically. The separation of mixed molecular populations is a pressing problem in single-particle analysis. Until recently, the task of distinguishing small structural variations was intractable, but developments in cryo-electron microscopy hardware and software now make it possible to address this problem. We have developed a general strategy for recognizing and separating structures of variable size from cryo-electron micrographs of single particles. The method uses a combination of statistical analysis and projection matching to multiple models. Identification of size variations by multivariate statistical analysis was used to do an initial separation of the data and generate starting models by angular reconstitution. Refinement was performed using alternate projection matching to models and angular reconstitution of the separated subsets. The approach has been successful at intermediate resolution, taking it within range of resolving secondary structure elements of proteins. Analysis of simulated and real data sets is used to illustrate the problems encountered and possible solutions. The strategy developed was used to resolve the structures of two forms of a small heat shock protein (Hsp26) that vary slightly in diameter and subunit packing. PMID- 14757058 TI - Location of auxilin within a clathrin cage. AB - The Dna J homologue, auxilin, acts as a co-chaperone for Hsc70 in the uncoating of clathrin-coated vesicles during endocytosis. Biochemical studies have aided understanding of the uncoating mechanism but until now there was no structural information on how auxilin interacts with the clathrin cage. Here we have determined the three-dimensional structure of a complex of auxilin with clathrin cages by cryo-electron microscopy and single particle analysis. We show that auxilin forms a discrete shell of density on the inside of the clathrin cage. Peptide competition assays confirm that a candidate clathrin box motif in auxilin, LLGLE, can bind to a clathrin construct containing the beta-propeller domain and also displace the well-characterised LLNLD clathrin box motif derived from the beta-adaptin hinge region. The means by which auxilin could both aid clathrin coat assembly and displace clathrin from AP2 during uncoating is discussed. PMID- 14757059 TI - Crystal structure of a cAMP-dependent protein kinase mutant at 1.26A: new insights into the catalytic mechanism. AB - The catalytic subunit of cAMP-dependent protein kinase has served as a paradigm for the entire kinase family. In the course of studying the structure-function relationship of the P+1 loop (Leu198-Leu205) of the kinase, we have solved the crystal structure of the Tyr204 to Ala mutant in complexes with Mg.ATP and an inhibitory peptide at 1.26A, with overall structure very similar to that of the wild-type protein. However, at the nucleotide binding site, ATP was found largely hydrolyzed, with the products ADP-PO(4) retained in the structure. High resolution refinement suggests that 26% of the molecules contain the intact ATP, whereas 74% have the hydrolyzed products. The observation of the substrate and product states in the same structure adds significant information to our understanding of the phosphoryl transfer process. Structural examination of the mutation site substantiates and extends the emerging concept that the hydrophobic core in the large lobe of the kinase might serve as a stable platform for anchoring key segments involved in catalysis. We propose that Tyr204 is critical for anchoring the P+1 loop to the core. Further analysis has highlighted two major connections between the P+1 loop and the catalytic loop (Arg165-Asn171). One emphasizes the hydrophobic packing of Tyr204 and Leu167 mediated through residues from the alphaF-helix, recently recognized as a signal integration motif, which together with the alphaE-helix forms the center of the hydrophobic core network. The other connection is mediated by the hydrogen bond interaction between Thr201 and Asp166, in a substrate-dependent manner. We speculate that the latter interaction may be important for the kinase to sense the presence of substrate and prepare itself for the catalytic reaction. Thus, the P+1 loop is not merely involved in substrate binding; it mediates the communication between substrate and catalytic residues. PMID- 14757060 TI - Cytochrome c and SDS: a molten globule protein with altered axial ligation. AB - Saccharomices cerevisiae (yeast iso-1) cytochrome c has been investigated in the presence of 100 mM SDS in order to simulate the interaction of cytochrome c with membrane. Under these circumstances, a high spin species with detached methionine axial ligand is observed through NMR, in analogy to findings on the horse heart protein. However, at variance with the latter system, for the yeast protein also a low spin species is detected, which appears to be present with a concentration of about 40% with respect to that of the high spin species. The R(1), R(2), [1H] 15N NOE of backbone amides which are not affected by paramagnetism are homogeneous and allow a simultaneous analysis of the data for the two species. The result is that the rotational correlation time is larger than in water and larger than expected on the basis of viscosity of the SDS-containing solution. This finding suggests interactions of cytochrome c with SDS. Furthermore, it appears that there is subnanosecond backbone mobility, which also accounts for the decreased intensity of NOE cross-peaks and may be associated with equilibria between helical and random coil structure. The dynamic behavior appears to be a common feature of the high spin and low spin species and is consistent with the presence of a molten globule state. The molten globule nature of the protein could account for the presence of the different axial coordination of the heme iron. Such findings are meaningful with respect to the physiology of cytochrome c as electron transfer protein and as promoter of apoptosis. PMID- 14757061 TI - Beyond the EX1 limit: probing the structure of high-energy states in protein unfolding. AB - Hydrogen exchange kinetics in native solvent conditions have been used to explore the conformational fluctuations of an immunoglobulin domain (CD2.domain1). The global folding/unfolding kinetics of the protein are unaltered between pH 4.5 and pH 9.5, allowing us to use the pH-dependence of amide hydrogen/deuterium exchange to characterise conformational states with energies up to 7.2kcal/mol higher than the folded ground state. The study was intended to search for discreet unfolding intermediates in this region of the energy spectrum, their presence being revealed by the concerted exchange behaviour of subsets of amide groups that become accessible at a given free energy, i.e. the spectrum would contain discreet groupings. Protection factors for 58 amide groups were measured across the pH range and the hydrogen-exchange energy profile is described. More interestingly, exchange behaviour could be grouped into three categories; the first two unremarkable, the third unexpected. (1) In 33 cases, amide exchange was dominated by rapid fluctuation, i.e. the free energy difference between the ground state and the rapidly accessed open state is sufficiently low that the contribution from crossing the unfolding barrier is negligible. (2) In 18 cases exchange is dominated by the global folding transition barrier across the whole pH range measured. The relationship between hydroxyl ion concentration and observed exchange rate is hyperbolic, with the limiting rate being that for global unfolding; the so-called EX1 limit. For these, the free energy difference between the folded ground state and any rapidly-accessed open state is too great for the proton to be exchanged through such fluctuations, even at the highest pH employed in this study. (3) For the third group, comprising five cases, we observe a behaviour that has not been described. In this group, as in category 2, the rate of exchange reaches a plateau; the EX1 limit. However, as the intrinsic exchange rate (k(int)) is increased, this limit is breached and the rate begins to rise again. This unintuitive behaviour does not result from pH instability, rather it is a consequence of amide groups experiencing two processes; rapid fluctuation of structure and crossing the global barrier for unfolding. The boundary at which the EX1 limit is overcome is determined by the equilibrium distribution of the fluctuating open and closed states (K(O/C)) and the rate constant for unfolding (k(u)). This critical boundary is reached when k(int)K(O/C)=k(u). Given that, in a simple transition state formalism: k(u)=K(#)k' (where K(#) describes the equilibrium distribution between the transition and ground state and k' describes the rate of a barrierless rearrangement), it follows that if the pH is raised to a level where k(int)=k', then the entire free energy spectrum from ground state to transition state could be sampled. PMID- 14757062 TI - Outlining folding nuclei in globular proteins. AB - Our theoretical approach for prediction of folding/unfolding nuclei in three dimensional protein structures is based on a search for free energy saddle points on networks of protein unfolding pathways. Under some approximations, this search is performed rapidly by dynamic programming and results in prediction of Phi values, which can be compared with those found experimentally. In this study, we optimize some details of the model (specifically, hydrogen atoms are taken into account in addition to heavy atoms), and compare the theoretically obtained and experimental Phi values (which characterize involvement of residues in folding nuclei) for all 17 proteins, where Phi values are now known for many residues. We show that the model provides good Phi value predictions for proteins whose structures have been determined by X-ray analysis (the average correlation coefficient is 0.65), with a more limited success for proteins whose structures have been determined by NMR techniques only (the average correlation coefficient is 0.34), and that the transition state free energies computed from the same model are in a good anticorrelation with logarithms of experimentally measured folding rates at mid-transition (the correlation coefficient is -0.73). PMID- 14757063 TI - Thermodynamic dissection of the binding energetics of proline-rich peptides to the Abl-SH3 domain: implications for rational ligand design. AB - The inhibition of the interactions between SH3 domains and their targets is emerging as a promising therapeutic strategy. To date, rational design of potent ligands for these domains has been hindered by the lack of understanding of the origins of the binding energy. We present here a complete thermodynamic analysis of the binding energetics of the p41 proline-rich decapeptide (APSYSPPPPP) to the SH3 domain of the c-Abl oncogene. Isothermal titration calorimetry experiments have revealed a thermodynamic signature for this interaction (very favourable enthalpic contributions opposed by an unfavourable binding entropy) inconsistent with the highly hydrophobic nature of the p41 ligand and the Abl-SH3 binding site. Our structural and thermodynamic analyses have led us to the conclusion, having once ruled out any possible ionization events or conformational changes coupled to the association, that the establishment of a complex hydrogen-bond network mediated by water molecules buried at the binding interface is responsible for the observed thermodynamic behaviour. The origin of the binding energetics for proline-rich ligands to the Abl-SH3 domain is further investigated by a comparative calorimetric analysis of a set of p41-related ligands. The striking effects upon the enthalpic and entropic contributions provoked by conservative substitutions at solvent-exposed positions in the ligand confirm the complexity of the interaction. The implications of these results for rational ligand design are discussed. PMID- 14757064 TI - Functional similarities and differences of an archaeal Hsp70(DnaK) stress protein compared with its homologue from the bacterium Escherichia coli. AB - Archaea are prokaryotes but some of their chaperoning systems resemble those of eukaryotes. Also, not all archaea possess the stress protein Hsp70(DnaK), in contrast with bacteria and eukaryotes, which possess it without any known exception. Further, the primary structure of the archaeal DnaK resembles more the bacterial than the eukaryotic homologues. The work reported here addresses two questions: Is the archaeal Hsp70 protein a chaperone, like its homologues in the other two phylogenetic domains? And, if so, is the chaperoning mechanism of bacterial or eukaryotic type? The data have shown that the DnaK protein of the archaeon Methanosarcina mazei functions efficiently as a chaperone in luciferase renaturation in vitro, and that it requires DnaJ, and the other bacterial-type chaperone, GrpE, to perform its function. The M. mazei DnaK chaperone activity was enhanced by interaction with the bacterial co-chaperone DnaJ, but not by the eukaryotic homologue HDJ-2. Both the bacterial GrpE and DnaJ stimulated the ATPase activity of the M. mazei DnaK. The M. mazei DnaK-dependent chaperoning pathway in vitro is similar to that of the bacterium Escherichia coli used for comparison. However, in vivo analyses indicate that there are also significant differences. The M. mazei dnaJ and grpE genes rescued E.coli mutants lacking these genes, but E.coli dnaK mutants were not complemented by the M. mazei dnaK gene. Thus, while the data from in vitro tests demonstrate functional similarities between the M. mazei and E.coli DnaK proteins, in vivo results indicate that, intracellularly, the chaperones from the two species differ. PMID- 14757065 TI - Identification of functionally important residues of Arp2/3 complex by analysis of homology models from diverse species. AB - We constructed homology models from the crystal structure of bovine Arp2/3 complex and sequences from six phylogenetically diverse species (Arabidopsis thaliana, Caenorhabditis elegans, Dictyostelium discoideum, Drosophila melanogaster, Saccharomyces cerevisiae, Schizosaccharomyces pombe) representing over 800 million years of evolution and used conserved surface residues to search for functionally important structural elements. The folds of the seven subunits and their core residues are well conserved, as well as residues at subunit interfaces. Only 45% of solvent-exposed surface residues are conserved and only 15% are identical across the seven species. Arp residues expected to interact with nucleotide and with the first and second actin subunits in a daughter filament are conserved and similar to actin. Arp residues required to form an Arp dimer differ from actin and may contribute to the dissociated state of the Arps in the unactivated complex. Conserved patches of surface residues guided us to candidate sites for nucleation promoting factors to interact with Arp3, Arp2, and ARPC3. Other conserved residues were used with experimental constraints to propose how residues on the subunits ARPC1, ARPC2, ARPC4 and ARPC5 might interact with the mother filament at branch junctions. PMID- 14757066 TI - A structural analysis of working memory and related cognitive skills in young children. AB - The aim of this study was to investigate the functional organisation of working memory and related cognitive abilities in young children. A sample of 633 children aged between 4 and 6 years were tested on measures of verbal short-term memory, complex memory span, sentence repetition, phonological awareness, and nonverbal ability. The measurement model that provided the best fit of the data incorporates constructs that correspond to the central executive, phonological loop, and episodic buffer subcomponents of working memory, plus distinct but associated constructs associated with phonological awareness and nonverbal ability. PMID- 14757067 TI - Age-related differences in the visual processes implied in perception and action: distance and location parameters. AB - The aim of the two present experiments was to examine the ontogenetic development of the dissociation between perception and action in children using the Duncker illusion. In this illusion, a moving background alters the perceived direction of target motion. Targets were held stationary while appearing to move in an induced displacement. In Experiment 1, 30 children aged 7, 9, and 12 years and 10 adults made a perceptual judgment or pointed as accurately as possible, with their index finger, to the last position of the target. The 7-year-old children were more perceptually deceived than the others by the Duncker illusion but there were no differences for the goal-directed pointing movements. In Experiment 2, 50 children aged 7, 8, 9, 10, and 11 years made a perceptual judgment or reproduced as accurately as possible, with a handle, the distance traveled by the target. Participants were perceptually deceived by the illusion, judging the target as moving although it was stationary. When reproducing the distance covered by the target, children were unaffected by the Duncker illusion. Our results suggest that the separation of the allocentric visual perception pathway from the egocentric action pathway occurs before 7 years of age. PMID- 14757068 TI - Variation among developmental dyslexics: evidence from a printed-word-learning task. AB - A word-learning task was used to investigate variation among developmental dyslexics classified as phonological and surface dyslexics. Dyslexic children and chronological age (CA)- and reading level (RL)-matched normal readers were taught to pronounce novel nonsense words such as veep. Words were assigned either a regular (e.g., "veep") or an irregular (e.g., "vip") pronunciation. Phonological dyslexics learned both regular and exception words more slowly than the normal readers and, unlike the other groups, did not show a regular-word advantage. Surface dyslexics also learned regular and exception words more slowly than the CA group, consistent with a specific problem in mastering arbitrary item-specific pronunciations, but their performance resembled that of the RL group. The results parallel earlier findings from Manis, Seidenberg, Doi, McBride-Chang, & Petersen [Cognition 58 (1996) 157-195] indicating that surface dyslexics and phonological dyslexics have a different profile of reading deficits, with surface dyslexics resembling younger normal readers and phonological dyslexics showing a specific phonological deficit. Models of reading and reading disability need to account for the heterogeneity in reading processes among dyslexic children. PMID- 14757069 TI - What makes the windows task difficult for young children: rule inference or rule use? AB - The windows task is difficult for young children. In this task, a child is shown two boxes with windows revealing that one is empty, whereas the other contains a treat. The child is asked to point to a box for an opponent to look in. The child then "wins" the contents of the other box (the treat). To pass the task, the child must use a rule such as "point to the empty box." But crucially, because the child is not told this rule by the experimenter, he or she must first infer it. Therefore, the windows task has two distinct requirements: (a) infer the rule "point to the empty box" and, once the rule is inferred, (b) use the rule by holding it in mind while inhibiting the prepotent response of pointing to the treat. In this study, the authors sought to determine which of these two requirements was responsible for poor performance on the windows task. They compared the performance of 3(1/2)-year-olds (N=40) on four tasks: the standard windows task, a version of the windows task that required rule use but not rule inference, and two versions of the day-night task that also required rule use but not rule inference. The relative performance on these four tasks and the pattern of correlations among them suggested that children had difficulty in inferring a rule that enables them to pass the task, whereas they had little difficulty in using the rule. Little evidence was obtained to suggest that the standard windows task requires inhibition. PMID- 14757070 TI - EXAFS study of mercury(II) sorption to Fe- and Al-(hydr)oxides. I. Effects of pH. AB - The study of mercury sorption products in model systems using appropriate in situ molecular-scale probes can provide detailed information on the modes of sorption at mineral/water interfaces. Such studies are essential for assessing the influence of sorption processes on the transport of Hg in contaminated natural systems. Macroscopic uptake of Hg(II) on goethite (alpha-FeOOH), gamma-alumina (gamma-Al(2)O(3)), and bayerite (beta-Al(OH)(3)) as a function of pH has been combined with Hg L(III)-edge EXAFS spectroscopy, FTIR spectroscopy, and bond valence analysis of possible sorption products to provide this type of information. Macroscopic uptake measurements show that Hg(II) sorbs strongly to fine-grained powders of synthetic goethite (Hg sorption density Gamma=0.39-0.42 micromol/m(2)) and bayerite (Gamma=0.39-0.44 micromol/m(2)), while sorbing more weakly to gamma-alumina (Gamma=0.04-0.13 micromol/m(2)). EXAFS spectroscopy on the sorption samples shows that the dominant mode of Hg sorption on these phases is as monodentate and bidentate inner-sphere complexes. The mode of Hg(II) sorption to goethite was similar over the pH range 4.3-7.4, as were those of Hg(II) sorption to bayerite over the pH range 5.1-7.9. Conversion of the gamma Al(2)O(3) sorbent to a bayerite-like phase in addition to the apparent reduction of Hg(II) to Hg(I), possibly by photoreduction during EXAFS data collection, resulted in enhanced Hg uptake from pH 5.2-7.8 and changes in the modes of sorption that correlate with the formation of the bayerite-like phase. Bond valence calculations are consistent with the sorption modes proposed from EXAFS analysis. EXAFS analysis of Hg(II) sorption products on a natural Fe oxyhydroxide precipitate and Al/Si-bearing flocculent material showed sorption products and modes of surface attachment similar to those for the model substrates, indicating that the model substrates are useful surrogates for the natural sediments. PMID- 14757071 TI - Experimental study of albumin and lysozyme adsorption onto acrylic acid (AA) and 2-hydroxyethyl methacrylate (HEMA) surfaces. AB - Many commercial soft contact lenses are based on poly-2-hydroxyethyl methacrylate (HEMA) and acrylic acid (AA) hydrogels. The adsorption of proteins, albumin and lysozyme, on such contact lens surfaces may cause problems in their applications. In this work the adsorption of proteins, albumin and lysozyme, on hydrogel surfaces, AA and HEMA, was investigated as a function of concentration of protein. Also the effects of pH and ionic strength of protein solution on the adsorption of protein were examined. The obtained results indicated that the degree of adsorption of protein increased with the concentration of protein, and the adsorption of albumin on HEMA surface at the studied pHs (6.2-8.6) was higher than AA surface, whereas the adsorption of lysozyme on AA surface at the same pHs was higher than HEMA. The change in ionic strength of protein solution affected the proteins adsorption on both AA and HEMA surfaces. Also, the amount of sodium ions deposited on the AA surface was much higher than HEMA surface. This effect can be related to the negative surface charge of AA and its higher tendency for adsorption of sodium ions compared to the HEMA surface. PMID- 14757072 TI - Exchange characteristics of monocarboxylic acids and monosulfonic acids onto anion-exchange resins. AB - In the previous paper (N. Kanazawa, K. Urano, N. Kokado, Y. Urushigawa, J. Colloid Interface Sci. 238 (2001) 196), the equilibria of propionic acid and benzoic acid adsorption onto three anion-exchange resins were investigated, and an equation was proposed that summed of the physical adsorption of the carboxylic acid molecule and the ion exchange of the dissociated carboxylate ion. The ion exchange equation, including a selectivity coefficient to chloride ion for each combination between carboxylate ions and anion-exchange resins, could be used in wide ranges of concentration and pH. In this research, ion-exchange equilibria using 16 anion-exchange resins and 9 organic acids including monocarboxylic and monosulfonic acids were investigated. It could be confirmed that the proposed equation applied to the ion exchange with these monoorganic acids. Characteristics of ion exchange between the organic anions and anion exchange were also studied by the selectivity coefficients. PMID- 14757073 TI - EXAFS studies on adsorption-desorption reversibility at manganese oxides-water interfaces. I. Irreversible adsorption of zinc onto manganite (gamma-MnOOH). AB - Microscopic structures of Zn(II) surface complexes adsorbed at the manganite (gamma-MnOOH)-water interface were studied using extended X-ray absorption fine structure (EXAFS) spectroscopy. Quantitative analysis of the first sphere showed that, in a 0.1 M NaNO(3) solution of pH 7.5, Zn(II) was adsorbed as a mixture of tetrahedral and octahedral structure (ZnO(4,6) polyhedra) and the average Zn-O distance was 2.00+/-0.01 A. EXAFS analysis of the second sphere showed that two typical atomic Zn-Mn distances of 3.07+/-0.01 and 3.52+/-0.02 A existed in the surface complexes, indicating that there were two types of linkage, i.e., the edge-linkage of high affinity and the corner-linkage of low affinity, between the ZnO(4,6) polyhedra and the MnO(6) octahedra of the manganite. Macroscopic adsorption-desorption experiments showed that adsorption of Zn(II) onto manganite was largely irreversible and the stronger edge-linkage mode was found to be responsible for the adsorption irreversibility. This result provided direct evidence from the molecular level for the basic hypothesis of the metastable equilibrium adsorption (MEA) theory that adsorption density is not a thermodynamic state variable because a given value of adsorption density could have different values of chemical potential, depending on the proportion between the edge and corner linkage modes. PMID- 14757074 TI - EXAFS studies on adsorption-desorption reversibility at manganese oxide-water interfaces. II. Reversible adsorption of zinc on delta-MnO2. AB - Microscopic structures of Zn(II) adsorbed at delta-MnO(2)-water interfaces were studied using extended X-ray absorption fine structure (EXAFS) spectroscopy. In a 0.1 M NaNO(3) solution of pH 5.50, hydrous Zn(II) was adsorbed onto the solid surface in the form of octahedral coordination. Adsorbed octahedral Zn(II) was located above and below the vacancy sites of delta-MnO(2). Each Zn was coordinated on one side to H(2)O molecules forming an H(2)O sheet and on the other side to oxygen atoms shared with layer MnO(6) octahedra forming a corner sharing octahedral interlayer complex. The average Zn-O and Zn-Mn distances were 2.07+/-0.01 and 3.52+/-0.01 A, respectively. Macroscopic adsorption-desorption isotherms showed that, in contrast to that of the Zn-gamma-MnOOH system, adsorption of Zn(II) on delta-MnO(2) was highly reversible. EXAFS results indicated that the highly reversible adsorption was due to the weak adsorption mode of the corner-sharing linkage between the adsorbate and adsorbent polyhedra. PMID- 14757075 TI - Size-dependent adsorption of 1,4-phenylenediisocyanide onto gold nanoparticle surfaces. AB - Adsorption of 1,4-phenylenediisocyanide (PDI) has been studied for different sized gold nanoparticles with mean diameters of 6, 14, 23, 40, 57, and 97 nm using UV-vis absorption spectroscopy and surface-enhanced Raman scattering (SERS). The SERS enhancement was found to be relatively weak for 6-nm particles due to less aggregation between PDI and gold particles. Concentration-dependent SERS spectra show that PDI was assumed to bridge two different gold particles at low concentrations of PDI, but as the concentration was increased, the bridge appeared to be broken, and PDI bonded to the gold particle only via one of its two isocyanide groups. For the 57- and 97-nm particles, however, the nu(NC)(free) stretching band in the SERS spectrum almost disappeared, even at a high bulk concentration of PDI, differently from the case of the smaller sizes (14, 23, and 40 nm). The 57- and 97-nm particles appeared to cross-link through the pendent isocyanide group even at a high bulk concentration. UV-vis absorption spectra indicated that PDI appeared to aggregate more extensively with increasing size in agreement with Raman data. Our result shows an example that the adsorption scheme of an aromatic diisocyanide may be varied depending on particle size as well as the bulk concentration. PMID- 14757076 TI - Isotherm analysis of phenol adsorption on polymeric adsorbents from nonaqueous solution. AB - Macroporous poly(methyl methacrylate-co-divinylbenzene) (PMMA), interpenetrating polymer adsorbent based on poly(styrene-co-divinylbenzene) (PS) and poly(methyl methacrylate-co-divinylbenzene) (PMMA/PS), and macroporous cross-linked poly(N-p vinylbenzyl acetylamide) (PVBA) were prepared for the adsorption of phenol from cyclohexane. The sorption isotherms of phenol on the three polymeric adsorbents were measured and fitted to Langmuir and Freundlich isotherms. It is shown that the Langmuir isotherm, which is based on a homogeneous surface model, is unsuitable to describe the sorption of phenol on the adsorbents from nonaqueous solution and the Freundlich equation fits the tested three adsorption systems well. The isosteric enthalpy was quantitatively correlated with the fractional loading for the sorption of phenol onto the three polymeric adsorbents. The surface energetic heterogeneity patterns of the adsorbents were described with functions of isosteric enthalpy. The results showed that the tested three polymeric adsorbents exhibited different surface energetic heterogeneity patterns. The initial isosteric enthalpy of phenol sorption on polymeric adsorbent has to do with the surface chemical composition and is free from the pore structure of the polymeric adsorbent matrix. Forming hydrogen bonds between phenol molecules and adsorbent is the main driving force of phenol sorption onto PVBA and PMMA adsorbent from nonaqueous solution. When phenol is adsorbed on PMMA/PS, pi-pi interaction resulting from the stacking of the benzene rings of the adsorbed phenol molecules and the pendant benzene ring of adsorbent is involved. PMID- 14757077 TI - Dispersion polymerization of acrylamide. III. Partial isopropyl ester of poly(vinyl methyl ether-alt-maleic anhydride) as a stabilizer. AB - Dispersion polymerization of acrylamide at 40 degrees C in t-butyl alcohol-water media of varying composition (50 to 90 vol% alcohol) using a partial isopropyl ester of poly(vinyl methyl ether-alt-maleic anhydride) as the stabilizer has been studied. The isopropyl ester derivative proves to be a better stabilizer than its precursor polymer. However, the particles were polydisperse in size. The number average diameter of the particles increases linearly with the solubility parameter of the dispersion medium. Dispersions of nanoparticles of polyacrylamide are produced when the t-butyl alcohol content of the dispersion medium is 90% by volume. Comparison of the particle diameter measured by transmission electron microscopy with that measured by dynamic light scattering in acetone reveals a thickness of the hairy stabilizer layer on the dispersion particles of 31 nm. PMID- 14757078 TI - Influence of substrate hydrophobicity on the adsorption of an amphiphilic diblock copolymer. AB - The adsorption of poly(tert-butylmethacrylate)-block-poly(2-(dimethylamino-ethyl) methacrylate) (PtBUMA-b-PDMAEMA) was studied by X-ray photoelectron spectroscopy (XPS) and atomic force microscopy (AFM) analysis performed on dried samples. The copolymer was dissolved in toluene at concentrations below (0.01 wt%) and above (0.05 and 1 wt%) the CMC; silicon (SiOH) and CH(3)-grafted silicon (SiCH(3)) were used as substrates. Whatever the concentration and the substrate, a layer of individual copolymer molecules, 1.5-3 nm thick, formed rapidly. The adsorbed amount was slightly higher and the resistance to AFM tip scraping was stronger on SiOH than on SiCH(3). This is attributed to hydrogen bonding between the PDMAEMA block and the OH groups of the silicon surface, leading to polarization of the adsorbed layer. Above the CMC, on SiOH, randomly scattered dot-like features (about 5 nm high) observed by AFM were attributed to individual micelles, which were not displaced by drying. On SiCH(3), the particles found on the top of the adsorbed layer were micelle aggregates, about 50 nm thick, the lateral size of which was strongly influenced by the rate of drying. This further difference between SiCH(3) and SiOH is tentatively attributed to the exposure of PDMAEMA by the adsorbed layer formed on SiCH(3), while only PtBUMA would be exposed by the layer adsorbed on SiOH. The red blood cell shape and the size of the micelles observed in single layers indicate that the PtBUMA corona was not made compact as a result of drying. PMID- 14757079 TI - Preparation and adsorption of refined polyelectrolyte complex nanoparticles. AB - We report on bulk and surface properties of centrifuged nonstoichiometric polyelectrolyte complex (PEC) dispersions. PECs were prepared by mixing poly(diallyldimethylammonium chloride) (PDADMAC) and sodium poly(maleic acid-co alpha-methylstyrene) (PMA-MS) at the monomolar mixing ratio of 0.6 and polymer concentration >/=1 mmol/l. Centrifugation of initial PEC dispersions revealed three phases: supernatant (SUP), coacervate (COAC), and an insoluble precipitate. Mass, turbidity, particle hydrodynamic radii (R(h)), and the titratable charge amount were determined for those phases. The turbid COAC phase consisted of 200 nm nanoparticles and carried 60% of the polymer mass and 20% of the titratable charge amount of the initial PEC dispersion. The SUP phase showed no turbidity and no such nanoparticles, but carried 80% of the initial titratable charge amount, presumably caused by excess polycations. Furthermore, linear dependences of turbidity and R(h) on COAC concentration was observed. COAC adsorption was studied at polyelectrolyte multilayer (PEM) modified silicon surfaces in dependence on both adsorption time and concentration using attenuated total reflection-Fourier transform infrared (ATR-FTIR) spectroscopy. The adsorption data were fitted by the simple Langmuir model. Comparison of COAC particles and polystyrene latices revealed similar adsorption features. SEM and AFM measurements resulted in hemispherically shaped adsorbed COAC particles with coverages >/=25%, whose calculated volumes correlated well with those in dispersion obtained by PCS. PMID- 14757080 TI - Nanoscale organization of adsorbed collagen: influence of substrate hydrophobicity and adsorption time. AB - The adsorption of collagen on polystyrene (PS) and polystyrene oxidized by oxygen plasma discharge (PSox) was studied as a function of time using radiolabeling, X ray photoelectron spectroscopy (XPS), and atomic force microscopy (AFM). Radiolabeling and XPS indicated that the initial step of adsorption was faster on PS than on PSox. AFM imaging under water revealed very different supramolecular organization of the adsorbed films depending on time and on the nature of the substrate: PS showed patterns of collagen aggregates at all adsorption times (from 1 min to 24 h); PSox was covered with a smooth layer except at long adsorption times (24 h), for which a mesh of collagen structures was observed. After fast drying, the collagen layer remained continuous and showed a morphology which recalled that observed under water. The mechanical stability of the adsorbed films was assessed under water by scraping with the AFM probe at different loading forces: no perturbations were created on PSox; in contrast, the layer adsorbed on PS was sensitive to scraping, the minimum force required to alter the collagen layer morphology increasing with time. These differences in the film properties were correlated with force measurements upon retraction: multiple adhesion forces were observed with collagen adsorbed on PS samples, whereas such an effect was never observed on PSox. The results show that the amount adsorbed and the organization of the adsorbed film respond differently to the adsorption time and that this is influenced by surface hydrophobicity. The quick initial adsorption on PS, compared to PSox, is thought to leave dangling collagen segments that are responsible for the observed morphology, for adhesion forces, and for lower mechanical resistance of the adsorbed layer. PMID- 14757082 TI - A kinetic investigation of the flocculation of alumina with polyacrylic acid. AB - Using a model colloidal system of alumina and polyacrylic acid (PAA), the kinetics of flocculation was investigated at low polymer concentrations and short durations (on the order of seconds). The polymer-induced flocculation processes obeyed Von Smoluchowski's bimolecular rate equation. Increases in the concentration of the polymer resulted in higher rate constants for the flocculation process. At a fixed concentration (say 50 ppb, parts per billion), the rate constant values showed a maximum value for 250,000 g mol(-1) polyacrylic acid. At this polymer concentration, calculations of the surface coverage of alumina by PAA molecules of different molecular weights show that for all the cases the coverage is nearly the same, approximately 1x10(-3), but the flocculation response and the rates are significantly different. This trend in flocculation characteristics is attributed to the critical polymer number density requirement for effective flocculation (at least partial charge neutralization and initiation of flocculation). The mechanism governing the flocculation at ultralow concentrations (50 ppb) is the synergistic effect of partial patch neutralization and bridging. PMID- 14757081 TI - Stabilization and encapsulation of human immunoglobulin G into biodegradable microspheres. AB - The instability of protein during preparation, storage, and release has become a major concern in recent years in the encapsulation of proteins into biodegradable polymers for controlled release systems. The present investigation was performed to study the mechanism of degradation of human immunoglobulin G (IgG) in double emulsion and solid-in-oil-in-water (S/O/W) encapsulation processes. The stabilizing effects of various excipients during the period of protein atomization using spray freeze-drying and subsequent encapsulation into polylactide-co-glycolide (PLGA) microspheres were explored. The size-exclusion high-performance liquid chromatography (SEC-HPLC) results showed that ultrasonication did not change the primary structure of IgG significantly. However, enzyme-linked immunosorbent assay (ELISA) revealed that the subsequent double-emulsion solvent evaporation process denatured nearly 80% of the total amount of IgG. This was possibly due to the adsorption, unfolding, and aggregation of IgG at the water/organic solvent interface. Both mannitol and trehalose could stabilize IgG during spray freeze-drying, with over 90% retention of its molecular integrity and immunoactivity, which were verified using SEC-HPLC and ELISA. Solid protein microparticles were further entrapped into monolithic type microspheres of PLGA using the S/O/W method. FTIR results suggested that the incomplete release that is often observed in the formulation of controlled protein release systems may be due to the degradation or aggregation of protein in the solid polymer matrix. PMID- 14757083 TI - Aggregate formation and collision efficiency in differential settling. AB - A new method of application of Stokesian dynamics, which can efficiently simulate movements of up to 500 particles with interparticle interactions in reasonable computational times, has been developed for the purpose of investigating particle cluster aggregation in aqueous systems. The method is applied to monodisperse non Brownian spherical particles aggregating in differential settling, while repulsive colloidal interaction is presumed to be negligible, so that a minimum separation distance can represent the attractive van der Waals force. The final aggregates formed by this algorithm, composed of 300 primary particles, have a common fractal dimension of approximately 2.0. The computed collision efficiency, defined as the product of a global and a capture efficiency, is about 5.77x10( 3). This value is significantly larger than the collision efficiency of primary particles colliding with an impermeable solid sphere of the same size as the aggregate, illustrating the important interplay between the permeability and the formation of aggregates. PMID- 14757084 TI - Synthesis and characterization of colloidal fluorescent mesoporous silica particles. AB - Mesoporous silica particles with narrow size distribution were obtained by a seeded growth process. Depending on the size of seeds and on the time of addition of reactants, the size of particles can be varied between 300 and 1000 nm. In a second step the dye fluorescein isothiocyanate can be embedded. The structure of these new silica particles with low density was investigated by SEM, XRD, BET, and confocal microscopy. PMID- 14757085 TI - Influence of dispersing agents and solution conditions on the solubility of crude kaolin. AB - Experiments measuring the solubility of kaolin particles in terms of the concentration of aluminum and silicon ions in supernatant were carried out as a function of the pH of the slurry over a wide range of dosages of different dispersing agents varying from 0.5 to 12 mg/(g solids). The concentrations of the metal ions in supernatant were found to be strongly affected by the type and the dosage of the dispersants and pH of the solution. In this study, the mechanism of the reaction between the dispersing agents and kaolin particles was studied and the dissolution capacities of metal ions (aluminum and silicon) were identified from kaolin particles in the absence and presence of dispersing agents. The three anionic dispersing agents used were sodium polyacrylate (Na-PAA), sodium hexametaphosphate (Na-HMP), and sodium silicate (Na-silicate), based on the industrial application of these agents and their ability to produce a stable dispersion for this purpose. PMID- 14757086 TI - Dendrimer-templated Ag-Pd bimetallic nanoparticles. AB - Ultrafine dendrimer-templated Ag-Pd bimetallic nanoparticles with various metal compositions have been prepared successfully using silver(I) bis(oxalato)palladate(II) complex. The use of an oxalate complex, in which two metal ions exist in one complex, is found to be effective in preventing unfavorable silver halide formation and thus suitable for the formation of Ag-Pd bimetallic nanoparticles. PMID- 14757087 TI - Physical properties of organic particulate UV-absorbers used in sunscreens. I. Determination of particle size with fiber-optic quasi-elastic light scattering (FOQELS), disc centrifugation, and laser diffractometry. AB - In this study microparticles consisting of a benzotriazole derivative, which are used as absorbers for UV radiation in cosmetic sunscreens, were investigated. The particles were micronized in presence of a dispersing agent by means of a ball milling process. According to the energy input different particle sizes were produced in the range of 0.16 to 4 microm. The particle sizes obtained after different stages of the micronization process were measured using fiber-optic quasi-elastic light scattering (FOQELS), disc centrifugation, and laser diffractometry. All methods showed satisfactory agreement over the whole range of sizes. With the FOQELS technique the particle size distribution could be resolved to sizes well below 0.1 microm. PMID- 14757088 TI - Temperature influence of nonionic polyethylene oxide and anionic polyacrylamide on flocculation and dewatering behavior of kaolinite dispersions. AB - Nonionic polyethylene oxide (PEO) and anionic polyacrylamide (PAM) flocculation of kaolinite dispersions has been investigated at pH 7.5 in the temperature range 20-60 degrees C. The surface chemistry (zeta potential), particle interactions (shear yield stress), and dewatering behavior were also examined. An increase in the magnitude of zeta potential of kaolinite particles, in the absence of flocculant and at a fixed PEO and PAM concentration, with increasing temperature was observed. The zeta potential behavior of the flocculated particles indicated a decrease in the adsorbed polymer layer thickness, while at the same time, however, the adsorbed polymer density showed a significant increase with increasing temperature. These results suggest that polymer adsorption was accompanied by temperature-influenced conformation changes. The hydrodynamic diameter and supernatant solution viscosity of both polymers decreased with increasing temperature, consistent with a change in polymer-solvent interactions and conformation, prior to adsorption. The analysis of the free energy (DeltaG(ads)) of adsorption showed a strong temperature dependence and the adsorption process to be more entropically than enthalpically driven. The polymer conformation change and increased negative charge at the kaolinite particle surface with increasing temperature resulted in decreased polymer bridging and flocculation performance. Consequently, the shear yield stress and the rate and the extent of dewatering (consolidation) of the pulp decreased significantly at higher temperatures (>40 degrees C). The temperature effect was more pronounced in the presence of PEO than PAM, with 40 and 20 degrees C indicated as the optima for enhanced performance of the latter and former flocculants, respectively. The results demonstrate that a temperature-induced conformation change, together with polymer structure type, plays an important role in flocculation and dewatering behavior of kaolinite dispersions. PMID- 14757089 TI - Quenching of fluorescence of 1-hydroxypyrene-3,6,8-trisulfonate (HPTS) by Cu2+, Co2+, Ni2+, I-, and cetylpyridinium (CP+) ions in water/AOT/heptane microemulsion. AB - The quenching of the fluorescence of HPTS (1-hydroxypyrene-3,6,8-trisulfonate) by Cu(2+), Ni(2+), Co(2+), I(-), and CP(+) (cetylpyridinium cation) has been studied in the w/o microemulsion medium formed with water, AOT [sodium salt of bis (2 ethylhexyl) sulfosuccinic acid], and heptane as components at two [H(2)O]/[AOT] ratios (omega), 6 and 20. The quenching process has been found to be dynamic in nature. The lifetimes of HPTS in the microemulsion medium in the absence and in the presence of quencher have been determined. The analysis of the results has been performed in terms of the Stern-Volmer equation and the quenching sphere of action model. The Poisson distribution equation has been also used in the analysis of the probability of quencher distribution in the microemulsion compartment. The quenching of HPTS has been found to be much lower in microemulsion than in bulk water. PMID- 14757090 TI - Characterization of the pores in hydrous ferric oxide aggregates formed by freezing and thawing. AB - Hydrous ferric oxides (HFO) are efficient sorbents for inorganic and organic pollutants and therefore have great potentials in environmental science and engineering applications. Freezing and thawing of HFO suspensions leads to the formation of dense HFO aggregates. It facilitates the handling and increases the drying rate of HFO. In this study, we used a combination of pycnometry, gas adsorption (N(2) gas, water vapor), and small-angle neutron scattering (SANS) to characterize the porosity and pore size distribution of dense HFO aggregates formed by freezing dialyzed HFO suspensions at -25 degrees C and thawing them at room temperature. The crystallinity of the HFO, which was a 2-line ferrihydrite, was not affected by this treatment. Wet sieving and laser diffraction analysis showed that the dense HFO aggregates had a unimodal size distribution with an average diameter of 235+/-35 microm. Increasing the freezing rate by cooling with liquid N(2) (-196 degrees C) resulted in much smaller aggregates with an average diameter of 20 microm. Adding NaNO(3) electrolyte to the HFO suspensions prior to freezing also resulted in the formation of smaller aggregates. The dense HFO aggregates formed at -25 degrees C had a porosity of 0.73+/-0.02 ll(-1). SANS revealed a unimodal size distribution of pores, with an average pore diameter of 2.0 nm. The diameter of the HFO crystallites was estimated by transmission electron microscopy to be 1.9+/-0.5 nm. Geometrical considerations taking into account the unit particle and average pore size suggest that the crystallites retain 1-2 layers of hydration water during the coagulation induced by freezing. Analysis by N(2) gas adsorption showed that drying the dense HFO aggregates induced a reduction in porosity by about 25% and shifted the pore size distribution to smaller diameters. Rewetting during water vapor adsorption did not induce significant changes of the aggregate structure. The specific surface area of the dry HFO aggregates was between 320 and 380 m(2)g(-1). PMID- 14757091 TI - Neodymium(III)-substituted bismuth titanate thin film generation using metal alkoxo, acyloxo, and beta-diketonato precursors employing a sol-gel route and using 4f-4f transition spectra as probes to explore kinetic performance. AB - Heterotrimetallic lanthanide-substituted bismuth titanate (BLT, where lanthanide is neodymium) with stoichiometry Nd(0.75)Bi(3.25)Ti(3)O(12) has been obtained in both highly homogenized crystalline and amorphous thin film forms using three different multicomponent precursors, A (formed from Nd(OC(3)H(7)(i))(3), Bi(OOC.CH(3))(3), and Ti(OC(3)H(7)(i))(4) taken in the desired stoichiometry), B (formed from Nd(OC(3)H(7)(i))(2)(acac), Bi(OOC.CH(3))(3), Ti(OC(3)H(7)(i))(3)(acac)), and C (formed from Nd(OC(3)H(7))(acac)(2), Bi(OOC.CH(3))(3), Ti(OC(3)H(7))(3)(acac), in the desired stoichiometry), and employing controlled acidic hydrolysis during the sol-gel method. Paramagnetic Nd(III), an f(3) metal ion, gives characteristic 4f-4f transition spectra in the visible and near infrared region. The sensitivity of 4f-4f transitions to minor coordination changes around paramagnetic Nd(3+) has been used to monitor hydrolysis during the progress of the sol-gel process of the multicomponent BLT precursors. The variation of intensities (oscillator strengths) of 4f-4f bands during hydrolysis, as well as the variation of Judd-Ofelt intensity parameters, has helped in following the preliminary kinetics of hydrolysis. Highly complex polycondensation reactions occurred during sol-gel hydrolysis of three BLT precursors. Rates of hydrolysis with respect to five 4f-4f transitions of Nd(III) were determined. The different types of multicomponent BLT precursors have shown different rates of hydrolysis, following the reactivity trend A > B > C. PMID- 14757092 TI - Surface characterization of poly(4-vinylpyridine) quaternized with tetradecyl bromide: effect of the degree of quaternization. AB - The surface behavior of poly(4-vinylpyridine) quaternized with tetradecyl bromide (P4VPC(14)) as function of the quaternization degree was studied. The percentage of vinylpyridine moieties quaternized was found to be 35 to 75%. Surface pressure area isotherms (pi-A) at the air-water interface were determined. The polymer monolayers show particular shapes at different quaternization degrees. In order to get information about the hydrophobicity degree of the polymeric systems, the surface energy (SE), and their dispersion and polar contributions, gamma(D) and gamma(P) respectively, measurements of the contact angle (CA) with water, bromobenzene, and cis-decalin were performed. The results obtained are dependent on the quaternization percentage of the functionalized polymers. At high quaternization degree, hysteresis in the pi-A diagrams was observed. The change in the P4VPC(14) molecular organization in the monolayer was also investigated during the compression and expansion processes. During these processes the monolayer was monitored by Brewster angle microscopy (BAM). For several cycles, all compression curves and all expansion curves showed a common intersection point. We have tried to describe the P4VPC(14) molecular organization at the air water interface by molecular dynamic simulation (MDS). The results are also discussed in terms of the effect of the counterions and water on the stability of the system over the more packed region. PMID- 14757093 TI - Effects of atmospheric plasma treatment on the interfacial characteristics of ethylene-vinyl acetate/polyurethane composites. AB - The surface characteristics of ethylene-vinyl acetate (EVA) were modified by argon, air, and oxygen plasma at atmospheric pressure. The surface energies of the EVA were evaluated by contact angles according to a sessile-drop method and adhesion energy (G(IC)) was estimated by a 180 degrees peel test with polyurethane (PU). After the plasma treatments, the surface free energies (or specific polar component) of the EVA increased about five times compared to that of virgin EVA. The adhesion between the EVA and the PU is significantly improved by the plasma treatment. Especially, Ar/air/O(2) plasma treatment increases G(IC) of EVA/PU up to about 600% compared to that of the sample using virgin EVA. PMID- 14757094 TI - Synthesis of novel N,N-di-n-alkylaldonamides and properties of their surface chemically pure adsorption layers at the air/water interface. AB - A homologous series of new surface-active N,N-di-n-alkyl-substituted amides derived from delta-D-gluconolactone and alpha-D-glucoheptonic-gamma-lactone were synthesized. The adsorption isotherms of their surface-chemically pure solutions were measured and evaluated to obtain the adsorption parameters of standard free energy of adsorption (DeltaG(0)(ad)), surface excess (Gamma( infinity )), cross sectional area of the adsorbed surfactant molecule (A(min)), and surface interaction parameter (H(s)). The surfactants possess comparatively low solubilities and do not form micelles at room temperature. This behavior is opposite to that of the other types of sugar surfactants showing excellent solubility and a strong tendency to association/micellization. The derivatives of gluconamide reveal surface activity slightly higher than that of the derivatives of glucoheptonamide, especially for long alkyl chains (n(C)>4). An increase in A(min) of about 6 A(2)/molecule for the gluconic series is observed. PMID- 14757095 TI - Detection of chemical alterations at internal walls of microchannel flow cells by nondestructive fluorescence depolarization. AB - A recent trend is the production of workable microchannel flow cells (MF cells). The nondestructive methods used to assess their reliability are based mainly on output monitoring and do not evaluate internal chemical interactions. We investigate a nondestructive method for evaluating changes in the chemical composition of the inner walls based on evaluation of the extent of alignment of a fluorescent probe in a liquid flowing within MF cells. Two MF cells were built with a 10-microm inner spacing. Their inner walls had four parallel SnO(2) strips, 2.00 mm wide, separated by 0.50-mm-wide glass strips. One cell had strips parallel to the flow and the other perpendicular. Flow-induced intermolecular alignment of rhodamine B in monoethylene glycol was scanned with 28-microm precision by fluorescence depolarization, using polarized-laser-induced fluorescence within induced flows (PLF-FI). No changes of polarization were seen when the flow was stopped. Under flowing conditions, polarization was always 4% lower in the glass region as compared to SnO(2). Glass had a higher solid-liquid interfacial tension (determined by contact angle measurements), thus being more wettable and increasing the drag, which propagates into the liquid flow, decreasing polarization. PLF-FI can thus identify regions with different chemical constitutions. PMID- 14757096 TI - Surface and micellar properties of new nonionic gemini aldonamide-type surfactants. AB - A new group of gemini aldonamide-type surfactants-N,N'-bisalkyl-N,N'-bis[(3 gluconylamide)propyl]ethylenediamines, N,N'-bisdodecyl-N,N'-bis[(3 glucoheptonylamide)propyl]ethylenediamine, and N,N'-bisalkyl-N,N'-bis[(3 lactobionylamide)propyl]ethylenediamines, (alkyl: n-C(8)H(17), n-C(12)H(25)), were synthesized and characterized. The surface properties, such as surface excess concentration, Gamma(cmc), surface area demand per molecule, A(min), efficiency in surface tension reduction, pC(20), the effectiveness of surface tension reduction, gamma(cmc), critical micelle concentration, cmc, and a measure of the tendency of the surfactant to adsorb at the aqueous/air interface relative to its tendency to form micelles in the bulk surfactant solution, cmc/C(20), and standard free energy of micellization, DeltaG(mic)(0), have been obtained by means of surface tension measurements. The standard fluorescence shift technique using PRODAN as a probe provide confirmation of the cmc values by an alternative method. Additionally, the micellar properties for the concentration near above the cmc have been characterized by the aggregation number, N(agg). The presence of the dimeric segments with the aldonamide hydrophilic units in the surfactant molecule is found to be the source of their unusual physicochemical behavior. They are very efficient at adsorbing at the free surface and at forming micelles in water. Their critical micelle concentration values are remarkably low. They reveal remarkably low A(min) values in relation to conventional nonionic surfactants, which is unexpected from the molecular dimensions for the molecule but which is possible if one assumes some type of multilayer structure or a coherent interfacial film. PMID- 14757097 TI - Asphaltene aggregation in organic solvents. AB - Asphaltenic solids formed in the Rangely field in the course of a carbon dioxide flood and heptane insolubles in the oil from the same field were used in this study. Four different solvents were used to dissolve the asphaltenes. Near infrared (NIR) spectroscopy was used to determine the onset of asphaltene precipitation by heptane titration. When the onset values were plotted versus asphaltene concentrations, distinct break points (called critical aggregation concentrations (CAC) in this paper) were observed. CACs for the field asphaltenes dissolved in toluene, trichloroethylene, tetrahydrofuran, and pyridine occurred at concentrations of 3.0, 3.7, 5.0, and 8.2 g/l, respectively. CACs are observed at similar concentrations as critical micelle concentrations (CMC) for the asphaltenes in the solvents employed and can be interpreted to be the points at which rates of asphaltene aggregations change. CMC values of asphaltenes determined from surface tension measurements (in pyridine and TCE) were slightly higher than the CAC values measured by NIR onset measurements. The CAC for heptane-insoluble asphaltenes in toluene was 3.1 g/l. Thermal gravimetric analysis (TGA) and elemental compositions of the two asphaltenes showed that the H/C ratio of the heptane-insoluble asphaltenes was higher and molecular weight (measured by vapor pressure osmometry) was lower. PMID- 14757098 TI - Emulsifying properties of acylated rapeseed (Brassica napus L.) peptides. AB - A peptide fraction having an average size of 5.6 amino acids has been purified from a rapeseed hydrolyzate, acylated using C(10)-C(14) acyl chlorides, and the surface tension values at the air-water interface and emulsifying properties studied. As compared with standard surface-active proteins, such as bovine serum albumin (BSA), and with detergents such as sodium dodecyl sulfate (SDS), acylated peptides exhibited particular surface characteristics. The surface tension at air water interface of acylated peptides ranged from 29.1 to 37.8 mN/m at equilibrium; these values were considerably lower than those for BSA and closer those for SDS, suggesting that acylated peptides pack at the air-water interface more like detergents than like proteins. The adsorption of acylated peptides to the oil-water interface was slower than for SDS or BSA, as deduced from the rather large size of oil droplets in emulsions (31-17 microm). Consequently, these emulsions creamed extensively during aging. Nevertheless, emulsions generated from acylated peptides were in general more stable to phase separation than those prepared from SDS. The C(14) acylated peptides were more effective for generating emulsions than the C(10) and C(12) derivatives, especially concerning the stability of emulsions against coalescence and phase separation, which was better than SDS and close to BSA. PMID- 14757099 TI - Mixed micelles of benzyldimethyltetradecylammonium chloride with tetradecyltrimethylammonium and tetradecyltriphenylphosphonium bromides: a head group contribution. AB - Mixed micelle formation by tetradecyltrimethylammonium (TTAB) and tetradecyltriphenylphosphonium bromides (TTPB) with benzyldimethyltetradecylammonium chloride (BTDACl) was studied with the help of conductivity and Kraft point measurements. The BTDACl + TTAB mixtures showed synergistic interactions whereas those of BTDACl + TTPB indicated weak antagonistic behavior. From Kraft temperature measurements, the enthalpy of fusion (H(1)(0)) from solid hydrated BTDACl to the liquid state in the presence of TTAB or TTPB was computed. It was found that DeltaH(1)(0) was much more positive for BTDACl + TTPB than for BTDACl + TTAB mixtures. PMID- 14757100 TI - PLURONIC x TETRONIC polyols: study of their properties and performance in the destabilization of emulsions formed in the petroleum industry. AB - In this work, a new family of branched poly(ethylene oxide-propylene oxide) (PEO PPO) block copolymers designed as TETRONIC polyols is evaluated and compared to linear PEO-PPO block copolymers designed as PLURONIC polyols. Additives have been employed as well in order to improve solubility of these materials in aqueous solution. Such additives include the sodium p-toluene sulfonate (NaPTS) hydrotrope and concentrated hydrochloric acid. Solubility tests and aqueous solution surface tension data showed consistent results: the structure of the block PEO-PPO copolymers exerts a huge influence on their solubility in water. The solubility of such copolymers is increased by the presence of the sodium toluene sulfonate (NaPTS) hydrotrope. The presence of HCl caused increased solubility for the copolymer TETRONIC polyol only, the effect being less than that observed for the hydrotrope. It is concluded that as regards emulsion stabilization, TETRONIC copolymer polyols perform better. Correlation between structure and properties leads to the optimization of block PEO-PPO copolymer selection aiming at using these materials for the separation of petroleum industry emulsions. PMID- 14757101 TI - Shear response of concentrated calcium carbonate suspensions. AB - The rheology of concentrated calcium carbonate suspensions is investigated with respect to addition of solution and dispersion polymers. System materials and composition are chosen to be similar, generically, to those in use in the coating of paper. Specifically, we investigate the particle volume fraction dependence of the relative viscosity, using both capillary and steady-shear concentric cylinder measurement methods to cover a broad range of concentrations. The results are interpreted in terms of semi-empirical models, such as the Krieger-Dougherty model. Oscillatory shear measurements are also employed to investigate the viscoelastic behavior of the concentrated suspensions. The measurements indicate that a common solution polymer thickener, carboxymethyl cellulose (CMC), causes depletion flocculation of calcium carbonate suspensions. PMID- 14757102 TI - In situ monitoring of metal nanoparticle self-assembly on protein-functionalized glass by broadband optical waveguide spectroscopy. AB - Broadband, time-resolved optical waveguide (OWG) spectroscopy has been used for in situ, real-time investigation into the self-assembly of metal nanoparticle monolayers. The OWG spectroscopy makes it possible to use the transverse electric (TE) and transverse magnetic (TM) modes to measure surface plasmon absorption of immobilized metal nanoparticles in two directions, parallel and normal to the waveguide surface. Therefore, this technique can provide direction-dependent information on the metal nanoparticles at the interface. In this paper, a 50 microm-thick glass plate was used as a slab waveguide and the kinetics of Au nanoparticle adsorption on a hemoglobin-functionalized glass substrate was examined in the early stage of self-assembly. The findings show that with the TE mode the surface plasmon resonance (SPR) behavior for immobilized Au nanoparticles is different from that with the TM mode. PMID- 14757103 TI - Can colloidal particles flocculate at two energy minima simultaneously in a quiescent medium? AB - The effect of electrostatic repulsion forces on the gravity-induced flocculation of non-Brownian particles in a quiescent medium is analyzed, and the phenomenon that colloidal particles can simultaneously flocculate at both the primary minimum and the secondary minimum of the total interaction curve of DLVO theory is proved. A narrow strip region describing this "combined" flocculation behavior exists on the stability diagram, and the concept of a limiting trajectory can be applied to explain this interesting result. PMID- 14757105 TI - Toxicology investigations with cell culture systems: 20 years after. AB - From almost 20 years the "in vitro" model has gained a wide ground in toxicological investigation, providing advanced tools, reliable protocols, mechanistic information. These advancements have been done thanks to different approaches, addressed at improving chemical testing and validating procedures, at exploring the cellular and molecular basis of toxicity, at studying the modifications that xenobiotics undergo in the cellular environment. In this review the most advanced cellular models, the mechanisms of cell death, the techniques to monitor gene activation, following chemical exposure, is highlighted. Moreover the more recent in vitro models to approach the biotransformation issue will be presented. PMID- 14757106 TI - Progressing toward the reduction, refinement and replacement of laboratory animal procedures: thoughts on some encounters with Dr Iain Purchase. AB - A variety of encounters with Dr Iain Purchase over the last 25 years are reviewed in relation to their significance in terms of progress toward the reduction, refinement and replacement of animal procedures in toxicology and toxicity testing. Included are the work of the first FRAME Toxicity Committee and the FRAME International Alternatives Validation Scheme, the International Conferences on Practical In Vitro Toxicology and the foundation of Toxicology in Vitro, the work of an Institute of Medical Ethics working party on issues raised by animal experimentation and alternative approaches, the need for in vitro assays for chemical carcinogenesis based on the transformation of human cells, the problem presented by the human hazard potential of thousands of chemicals already in use before modern regulations for the registration of new chemicals came into force, and the importance of testing strategies with a focus on the integrated use of non-animal computer-based and in vitro test systems. PMID- 14757107 TI - A theoretical model for simulating the outcome of mechanism based in vitro toxicity testing strategies. AB - In order to investigate the fundamental principles that influence the optimal selection of toxicity test methods for the evaluation of chemical hazards, it is useful to have a design model to explore possible alternative testing strategies. In general, our lack of detailed knowledge of the mechanisms of toxicity (including not only the early events in the interaction of chemicals with biological systems, but the sequence of events that lead to experimentally measurable toxicity) limits the development of realistic dynamic models of the toxicological process for individual chemicals. We report here the development of a theoretical model that includes two independent hypothetical mechanisms of toxicity. The mechanisms are designed to be qualitatively similar to known mechanisms of action. The model is exercised to simulate the experimental data that would be obtained for a collection of "test" and "validation" chemicals using a single or a combination of two toxicity tests. The data generated are used to evaluate the "relevance" of the testing strategy based on the two proposed in vitro toxicity tests. PMID- 14757108 TI - Metabolism of nicotine and induction of CYP1A forms in precision-cut rat liver and lung slices. AB - The aim of this study was to investigate xenobiotic metabolism and induction of cytochrome P450 (CYP) forms in precision-cut rat liver and lung slices, employing nicotine as a model compound. Freshly cut rat liver and lung slices metabolised nicotine to the major metabolite cotinine. Observed Km values for cotinine formation in liver and lung slices were 323 and 41.7 microM, respectively, with corresponding V(max) values of 47.2 and 3.21 pmol/min/mg protein, respectively. Rat liver and lung slices were cultured for 48 h with Aroclor 1254, benzo(a)pyrene, nicotine and cotinine. Both Aroclor 1254 and benzo(a)pyrene produced a marked induction of CYP1A-dependent 7-ethoxyresorufin O-deethylase activity in both liver and lung slices. However, while nicotine induced 7 ethoxyresorufin O-deethylase activity in lung slices, but not in liver slices, cotinine did not induce enzyme activity in either liver or lung slices. Overall, while higher rates of nicotine metabolism were observed in rat liver slices, nicotine-induced CYP1A form induction was observed in lung slices. These results demonstrate the usefulness of precision-cut tissue slices for studying tissue differences in xenobiotic metabolism and CYP form induction. PMID- 14757109 TI - Activation of signalling pathways during hepatocyte isolation: relevance to toxicology in vitro. AB - The "Holy Grail" of in vitro toxicology is to develop assay systems that mimic the in vivo situation and hence reduce the need for toxicity tests employing experimental animals. However a major problem to be overcome with cell culture models is the rapid loss of differentiated phenotype that markedly limits extrapolation of results to the whole animal (i.e. human) situation. This limitation is most obvious in the application of hepatocyte cultures to predict pathways of metabolism mediated toxicity and results from the rapid loss of cytochrome P450 content. Here we demonstrate that changes in hepatocyte gene expression (e.g. MAP kinase and NF-kappaB activation) occur very early into the well established hepatocyte isolation procedure employing collagenase suggesting that hepatocytes are undergoing a pro-inflammatory ('acute phase') response before they are cultured. Data is presented indicating that the stimulus is, in part, due to oxidative stress but the demonstration of endotoxins in collagenase preparations is likely to exacerbate the situation. Thus appreciation of these early events during hepatocyte isolation represents the surest foundation for the successful application of cultured hepatocytes to toxicology rather than relying on traditional manipulations of hepatocyte culture medium/substratum once differentiated phenotype has already been lost. PMID- 14757110 TI - Dendritic cells and skin sensitisation hazard assessment. AB - Allergic contact dermatitis is an important occupational and environmental health disease. There is a need, therefore, to identify skin sensitisation hazard, and to assess accurately likely risks to human health. During the past 15 years very significant advances have been made in our understanding of the cellular and molecular mechanisms that serve to initiate and regulate cutaneous immune responses, including the acquisition of skin sensitisation. This has facilitated parallel advances in the identification and characterisation of skin sensitising chemicals and the development of more robust approaches to risk assessment. It is relevant to consider whether advances in immunobiology provide opportunities also for the design of alternative approaches to the toxicological evaluation of skin sensitisation, including the development of in vitro methods. Here we review the potential use of strategies based on analysis of responses induced in Langerhans cells and dendritic cells; professional antigen processing and presenting cells that are known to play pivotal roles during the induction phase of adaptive immune responses. PMID- 14757111 TI - Release of inflammatory cytokines, cell toxicity and apoptosis in epithelial lung cells after exposure to ambient air particles of different size fractions. AB - Several studies have shown that particles of smaller size may be more potent than larger to induce inflammatory and toxic responses in cultured lung cells. However, the relative importance of different size fractions of ambient PM to induce such effects is still not known. In this study, we investigated the potency of different size fractions of urban ambient air particles to induce release of inflammatory cytokines in the human alveolar cell line A549 and primary rat type 2 cells. A mineral-rich ambient air PM10 sample collected in a road tunnel (road PM10) was also included. The coarse fraction of the urban ambient air particles demonstrated a similar or higher potency to induce release of the proinflammatory cytokines IL-8/MIP-2 and IL-6 compared to the fine and ultrafine fractions. The coarse fraction was also the most toxic in both cell systems. In contrast to the A549 cells, no induction of cytokine release was induced by the ultrafine particles in the primary type 2 cells. The mineral-rich road PM10 may be equally or more potent than the various size fractions of the ambient air particles to induce cytokines in both cell types. In conclusion, the coarse fraction of ambient particles may be at least as potent by mass as smaller fractions to induce inflammatory and toxic effects in lung cells. PMID- 14757112 TI - The testing of chemicals in the Syrian hamster embryo (SHE) cell transformation assay for assessment of carcinogenic potential. AB - The Syrian hamster embryo (SHE) cell transformation assay was used to test 28 chemical substances for their ability to induce morphologically transformed colonies. The purpose was to determine how well the assay method could be transferred from an experienced laboratory by including 18 chemicals previously evaluated and 10 new chemicals. Technical training was obtained in the experienced lab prior to testing. The assay was conducted at pH 6.7, a treatment period of 7 days was used, and single experiments were performed for each chemical. With this limited testing, 78% concordance with rodent bioassay results was obtained, and this high concordance would have increased if small, but statistically negative responses from single trials were overturned by positive data from repeat trials. Similarly, the results were highly concordant (90%) with the experienced lab results; only 2 chemical evaluations were discordant, and the use of repeat experiments would likely have eliminated these apparent disagreements. Thus, with appropriate training, the pH 6.7 SHE assay was successfully and reliably transferred. PMID- 14757113 TI - The significance of in vitro rat skin absorption studies to human risk assessment. AB - The present paper reviews the comparative rates of skin penetration between rat and man for a total of 14 chemicals in in vitro absorption studies. The results showed that in vitro absorption assays are capable of demonstrating large differences in the rate of skin penetration. Saturation of absorption was also frequently observed at higher exposure levels. The highest absorption rates through rat and human epidermis were observed with compounds with a molecular weight of approximately 300, an aqueous solubility of approximately 1-6 mg/l, and a log10 (P(OCTANOL/WATER)) of approximately 3-4. When the absorption data for 3 compounds with a log10 (P(OCTANOL/WATER)) of 2.9-3.0 were compared, there appeared to be an inverse relationship between molecular weight/aqueous solubility and the rate of dermal absorption. Lipophilic compounds with low aqueous solubility (<4 mg/l) showed the highest penetration rates through rat skin, but this was not always the case for human skin. The human skin was invariably less permeable to all tested substances than rat skin, though no constant factor of difference could be identified. The factor of difference would not appear to be determined by molecular weight, lipophilicity, or aqueous solubility. The actual systemic exposure of humans may be significantly overestimated if risk assessment is based only on the results of an in vivo rat study. It would appear that dermal penetration through human skin should be based on the combined use of in vivo and in vitro data, using the following equation: %Human dermal penetration= [[% dermal penetration rat (in vivo)] x [rate dermal penetration human (in vitro)]] / [rate dermal penetration rat (in vitro)] PMID- 14757114 TI - Acute systemic toxicity--prospects for tiered testing strategies. AB - After many years of controversy and debate, the LD50 test was finally deleted by the end of 2002. Three alternative animal tests, the Fixed Dose Procedure, the Acute Toxic Class Method and the Up and Down Procedure have been developed which give rise to significant improvements in animal welfare. They have recently undergone revision to improve their scientific performance but more importantly to increase their regulatory acceptance. They can now be used within a strategy for acute toxicity testing for all types of test substances and for all regulatory and in-house purposes. In vitro cytotoxicity tests could be used as adjuncts to these alternative animal tests within the next year or so to improve dose level selection and thus give further modest improvements in the numbers of animals used. However, the total replacement of animal tests requires a considerable amount of further test development, followed by validation, and is at least 10 years away. PMID- 14757115 TI - The GABAergic phenotype of the "glutamatergic" granule cells of the dentate gyrus. AB - The granule cells of the dentate gyrus (DG), origin of the mossy fibers (MFs), have been considered to be glutamatergic. However, data obtained with different experimental approaches in recent years may be calling for a redefinition of their phenotype. Although they indeed release glutamate for fast neurotransmission, immunohistological and molecular biology evidence has revealed that these glutamatergic cells also express GABAergic markers. The granule cell expression of a GABAergic phenotype is developmentally regulated. Electrophysiological studies reveal that during the first 3 weeks of age, mossy fiber stimulation provokes monosynaptic fast inhibitory transmission mediated by GABA, besides the monosynaptic excitatory glutamatergic transmission, onto their targets in CA3. After this age, mossy fiber GABAergic transmission abruptly disappears and the GABAergic markers are undetected. In the adult, the GABAergic markers are upregulated and GABA-mediated transmission emerges after induction of hyperexcitability. The simultaneous glutamate- and GABA-mediated signals share the same plastic and pharmacological characteristics that correspond to neurotransmission of mossy fiber origin. This intriguing evidence gives rise to two fundamental points of discussion. The first is the plausible fact that glutamate and GABA, two neurotransmitters of opposing actions, are coreleased from the mossy fibers. The second relates to its functional implications that can be immediately inferred, as the dentate gyrus can exert direct GABA-mediated excitatory actions early in life and inhibitory actions in young and adult hippocampus. This evidence poses the need to reevaluate and reinterpret some aspects of the physiology of the mossy fiber pathway under normal and pathological conditions. This work reviews the recent evidence that supports the assumption that glutamate and GABA can be coreleased from a single pathway, the mossy fibers, and makes some considerations about its functional implications. PMID- 14757116 TI - Functional organisation of central cardiovascular pathways: studies using c-fos gene expression. AB - Until about 10 years ago, knowledge of the functional organisation of the central pathways that subserve cardiovascular responses to homeostatic challenges and other stressors was based almost entirely on studies in anaesthetised animals. More recently, however, many studies have used the method of the expression of immediate early genes, particularly the c-fos gene, to identify populations of central neurons that are activated by such challenges in conscious animals. In this review we first consider the advantages and limitations of this method. Then, we discuss how the application of the method of immediate early gene expression, when used alone or in combination with other methods, has contributed to our understanding of the central mechanisms that regulate the autonomic and neuroendocrine response to various cardiovascular challenges (e.g., hypotension, hypoxia, hypovolemia, and other stressors) as they operate in the conscious state. In general, the results of studies of central cardiovascular pathways using immediate early gene expression are consistent with previous studies in anaesthetised animals, but in addition have revealed other previously unrecognised pathways that also contribute to cardiovascular regulation. Finally, we briefly consider recent evidence indicating that immediate early gene expression can modify the functional properties of central cardiovascular neurons, and the possible significance of this in producing long-term changes in the regulation of the cardiovascular system both in normal and pathological conditions. PMID- 14757117 TI - Neurodegeneration in familial amyloid polyneuropathy: from pathology to molecular signaling. AB - Familial amyloid polyneuropathy (FAP) is an autosomal dominant neurodegenerative disorder related to the systemic deposition of mutated transthyretin (TTR) amyloid fibrils, particularly in peripheral nervous system (PNS). TTR fibrils are diffusely distributed in the PNS of FAP patients, involving nerve trunks, plexuses and ganglia. In peripheral nerves, amyloid deposits are prominent in the endoneurium, near blood vessels, Schwann cells and collagen fibrils. Fiber degeneration is axonal, beginning in the unmyelinated and low diameter myelinated fibers. Several hypotheses have been raised to explain axonal and neuronal loss: (i) compression of the nervous tissue by amyloid; however, a cause-effect relationship between amyloid deposition, structural nerve changes and degeneration was never clearly made; (ii) role of nerve ischemia secondary to lesions caused by perivascular amyloid, which is also doubtful as compromised blood flow was never demonstrated; (iii) lesions in the dorsal root ganglia neurons or Schwann cells. Recently, evidence for the presence of toxic non fibrillar TTR aggregates early in FAP nerves constituted a first step to unravel molecular signaling related to neurodegeneration in FAP. The toxic nature of TTR non-fibrillar aggregates, and not mature TTR fibrils, was evidenced by their ability to induce the expression of oxidative stress and inflammation-related molecules in neuronal cells, driving them into apoptotic pathways. How these TTR aggregates exert their effects is debatable; interaction with cellular receptors, namely, the receptor for advanced glycation endproducts (RAGE), is a probable candidate mechanism. The pathology and the yet unknown molecular signaling mechanisms responsible for neurodegeneration in FAP are discussed. PMID- 14757118 TI - Cerivastatin inhibits proliferation of interleukin-1 beta-induced rat mesangial cells by enhanced formation of nitric oxide. AB - The antiproliferative effect of statins on mesangial cells could represent a new therapeutic approach in glomerulonephritis. We studied in rat mesangial cells whether the antiproliferative action of cerivastatin on mesangial cells may be mediated by mesangial nitric oxide (NO) formation due to the inducible NO synthase (iNOS) or by induction of cyclooxygenase-2. Mesangial cells were stimulated with interleukin-1 beta and treated with cerivastatin for 24 h. Cell proliferation was examined by bromodeoxy-uridine (BrdU) incorporation, and nitrite and prostaglandin production was measured in supernatants as a means for iNOS or cyclooxygenase-2 activity. iNOS and cyclooxygenase-2 expression was quantified by Northern and Western blot analyses. Cerivastatin (0.0625 microM) significantly inhibited DNA synthesis in interleukin-1 beta-stimulated mesangial cells without altering cell viability. Interleukin-1 beta-induced nitrite production was twofold increased by 0.05 microM cerivastatin, and this effect could be reversed by addition of 100 microM mevalonate. iNOS mRNA levels increased sixfold (33% of maximum) in cerivastatin-treated mesangial cells as compared with vehicle-treated controls (3.5% of maximum). iNOS and cyclooxygenase 2 protein expression increased threefold (iNOS: 2.77+/-0.53/cyclooxygenase-2: 3.49+/-1.25). The NOS inhibitors N-methyl-L-arginine (L-NMMA) and L-N6-(1 iminoethyl)lysine (L-NIL) reversed the antiproliferative effect of cerivastatin. The cyclooxygenase-2 inhibitor celecoxib did not alter DNA synthesis and iNOS or cyclooxygenase-2 expression, but blocked prostacyclin production in interleukin-1 beta and cerivastatin-treated mesangial cells. In conclusion, cerivastatin increased cytokine-induced iNOS and cyclooxygenase-2 expression, thus constituting NO-regulated growth inhibition of mesangial cells. PMID- 14757119 TI - Yohimbine inhibits firing activities of rat dorsal root ganglion neurons by blocking Na+ channels and vanilloid VR1 receptors. AB - Yohimbine, an indole alkaloid, is a natural alpha(2)-adrenoceptor antagonist and is frequently used to assess the mechanism of a drug's effect on alpha adrenoceptors. Recently, several studies showed that yohimbine exhibited analgesic effects in in vivo animal models. However, the underlying mechanism is not known. We investigated the effects of yohimbine on Na(+) channels and vanilloid VR1 receptors in dorsal root ganglion cells. We found that yohimbine inhibited tetrodotoxin-sensitive Na(+) channels (Na(V)1.2), the tetrodotoxin resistant Na(+) channels, including both slow inactivating (Na(V)1.8) and persistent (Na(V)1.9) Na(+) channels, and capsaicin-sensitive vanilloid VR1 receptors. Action potential firing activities of dorsal root ganglion neurons evoked by current injection or capsaicin were eliminated by yohimbine. The blocking effects of yohimbine on nociceptive-related ion channels and firing activities of dorsal root ganglion neurons may underlie the ionic mechanism of yohimbine's analgesic effects observed in in vivo studies. PMID- 14757120 TI - Differentiation-inducing factor-1-induced growth arrest of K562 leukemia cells involves the reduction of ERK1/2 activity. AB - The differentiation-inducing factor-1 (DIF-1) is a signal molecule that induces stalk cell differentiation in the cellular slime mold Dictyostelium discoideum. In addition, DIF-1 is a potent antileukemic agent that induces growth arrest in K562 cells. In this study, we investigated the mechanism of action of DIF-1 in K562 cells in the light of cell-cycle regulators such as cyclins, retinoblastoma protein (pRb), and the mitogen-activated protein kinase (MAPK) family. DIF-1 down regulated cyclins D/E and a phosphorylated form of pRb (p-pRb), and thereby induced G(1) arrest of the cell cycle. DIF-1 inactivated the extracellular signal regulated kinase (ERK) in a biphasic manner but did not affect the c-Jun N terminal kinase (JNK) or p38 MAPK. The MEK (MAPK kinase) inhibitor, U0126, which has been shown to induce growth arrest, inactivated ERK and down-regulated cyclins D and E. Although DIF-1 activated the phosphatidylinositol 3-kinase (PI 3K)/Akt pathway, neither wortmannin nor 2-(4-morpholinyl)-8-phenyl-4H-1 benzopyran-4-one (LY294002; PI-3K inhibitors) cancelled DIF-1-induced growth arrest. The present results suggest that ERK inactivation may be involved in DIF 1-induced growth arrest and that PI-3K activity is not required for DIF-1-induced growth arrest in K562 cells. PMID- 14757121 TI - Electrophysiologic properties of lidocaine, cocaine, and n-3 fatty-acids block of cardiac Na+ channels. AB - Lidocaine and cocaine, two local anesthetics, and n-3 polyunsaturated fatty acids in fish oils, inhibit the voltage-gated Na(+) channels of cardiomyocytes. This inhibition by lidocaine and n-3 fish oil is associated with antiarrhythmic effects, whereas with cocaine lethal arrhythmias may occur. These electrophysiologic studies show that at the concentrations tested, the n-3 fish oil fatty acids and lidocaine share three actions on I(Na): a potent inhibition of I(Na); a strong voltage-dependence of this inhibition; and a large shift of the steady-state inactivation to hyperpolarized potentials. By contrast cocaine shares only the potent inhibition of I(Na). The voltage-dependence of the inhibition is much decreased with cocaine, which produces only a very small leftward shift of the voltage-dependence of inactivation. The large leftward shift of the steady-state inactivation seems very important in the prevention of fatal arrhythmias by the n-3 fatty acids. Thus, we suggest that it is lack of this effect by cocaine, which is one factor, that eliminates its ability to prevent fatal cardiac arrhythmias. Further we report that in cultured neonatal rat cardiomyocytes n-3 fish oil fatty acids terminate the tachycardia induced by the alpha(1) adrenergic agonist, phenylephrine, whereas cocaine accelerates the tachycardia and causes bouts of tachyarrhythmias. PMID- 14757122 TI - A four-ligand hypercube model to quantify allosteric interactions within the GABAA receptor complex. AB - The aim of this study was to investigate the couplings between various binding sites on the GABA(A) receptor complex. We investigated combinations of three test compounds: (1) GABA (gamma-aminobutyric acid), (2) Org 20549 [(2 beta 3 alpha 5 alpha)-21hydroxy-3Hydroxy-2(4morpholinyl)pregnan-20one methane-sulphonate)], a neuroactive steroid and (3) retigabine (D-23129, N-(2-amino-4-(4 fluorobenzylamino)-phenyl) carbamic acid ethyl ester), a new antiepileptic drug. Receptor-binding assays were conducted using rat brain membranes. [3H]TBOB ([3H] t-butyl-bicyclo-ortho-benzoate) was the tracer ligand. All three test compounds inhibited the binding of [3H]TBOB with EC(50)'s of 4.0, 98 and 23 microM, respectively. Isobolic analysis of the combination data showed that the three compounds act in synergy in displacing [3H]TBOB. These interactions could be described and quantified by a hypercube model in which each of the three test compounds and [3H]TBOB bind to different, allosterically coupled sites such that each of the test compounds allosterically displaces the tracer [3H]TBOB and allosterically enhances the affinity of any other test compound by a factor 4.4. The simultaneous binding of any two ligands enhances the affinity of the third by a factor 9. These results may contribute to the understanding of individual variability in drug responses and to the discussion about rational polytherapy. PMID- 14757123 TI - Multiple effects of arginine vasopressin on prostaglandin E2 synthesis in fibroblasts. AB - Recent evidence supports the viewpoint that vasopressin, a neurohypophyseal peptide, should be also considered as a neuroendocrine modulator of immune and inflammatory responses. In this work we investigated the role of vasopressin in the regulation of prostaglandin E(2) synthesis by human dermal fibroblasts. Recombinant human interleukin-1 beta increased prostaglandin E(2) synthesis in fibroblasts about sixfold. The prostaglandin E(2) response to interleukin-1 beta was attenuated by lower concentrations of vasopressin (10(-10)-10(-9) M). By contrast, higher concentrations (10(-8)-10(-7) M) of vasopressin effected significant enhancement of the interleukin-1 beta-induced prostaglandin E(2) synthesis. In a similar way, vasopressin (10(-8)-10(-7) M), in the absence of interleukin-1, significantly increased prostaglandin E(2) production. An inhibitory effect of lower concentrations of vasopressin was also observed on basal production of prostaglandin E(2). The effects of vasopressin on basal and interleukin-1 beta-induced prostaglandin E(2) synthesis were antagonized by selective vasopressin receptor antagonists. The findings presented here disclose a novel modulatory role of vasopressin on prostaglandin E(2) synthesis in human dermal fibroblasts and suggest a possible role of vasopressin in the regulation of inflammation. PMID- 14757124 TI - Peroxovanadate induces alpha 1B-adrenoceptor phosphorylation and association with protein kinase C. AB - Peroxovanadate induced a marked increase in the phosphorylation state of alpha(1B)-adrenoceptors. The effect was dose-dependent (EC(50) approximately 2 microM) and rapid, reaching its maximum in 5 min and remaining at this level for 30 min. Hydrogen peroxide also increased alpha(1B)-adrenoceptor phosphorylation but to a lesser extent, in an ephemeral fashion, and only at high (millimolar) concentrations. The effect of peroxovanadate was blocked by inhibitors of protein kinase C such as staurosporine and rottlerin and only partially reduced by genistein and inhibitors of phosphoinositide 3-kinase. Protein kinase C alpha, delta and epsilon are associated with the alpha(1B)-adrenoceptor under basal conditions, as reflected by coimmunoprecipitation. Such association was increased by peroxovanadate for all isoforms. In contrast, hydrogen peroxide increased only the association of the epsilon isoform to the adrenoceptor. Peroxovanadate decreased the ability of noradrenaline to increase intracellular calcium, indicating that the receptor phosphorylation induced has functional consequences. PMID- 14757125 TI - Cytochrome P450 isoenzymes involved in rat liver microsomal metabolism of californine and protopine. AB - Studies are described on the cytochrome P450 (CYP) isoenzyme dependence of the main metabolic steps of the Eschscholtzia californica alkaloids californine and protopine using rat liver microsomes. Preparations of E. californica are in use as phytopharmaceuticals and as herbal drugs of abuse. CYP isoenzyme dependences were studied using specific chemical inhibitors for CYP1A2, CYP2D1, and CYP3A2 (alpha-naphthoflavone, quinine, and ketoconazole, respectively). CYP2C11 was inhibited by specific antibodies for lack of specific chemical inhibitors. Californine N-demethylation was mainly catalyzed by CYP3A2 and to a minor extent by CYP1A2 and CYP2D1, but not by CYP2C11. CYP2D1 and CYP2C11 were shown to be mainly involved in demethylenation of both, californine and protopine, while CYP1A2 and CYP3A2 showed only minor contribution. Kinetic parameters of the reactions were established. K(m) and V(max) values for the californine N demethylation were 4.5+/-4.7 microM and 22.9+/-13.7 min/mg protein (high affinity) and 161.3+/-16.7 microM and 311.8+/-39.4 min/mg protein (low affinity), respectively. Californine demethylenation and protopine demethylenation showed substrate inhibition and K(m) and V(max) values were 5.0+/-0.5 and 7.1+/-0.6 microM and 83.3+/-2.6 and 160.7+/-4.0 min/mg protein, respectively. PMID- 14757126 TI - 5-Aza-2'-deoxycytidine and paclitaxel inhibit inducible nitric oxide synthase activation in fibrosarcoma cells. AB - Given the important role of gaseous free radical nitric oxide (NO) in tumor cell biology, we investigated the ability of the anti-cancer drugs 5-Aza-2' deoxycytidine (ADC) and paclitaxel to modulate NO production in mouse L929 fibrosarcoma cells. Both drugs reduced IFN-gamma-stimulated NO release in cultures of L929 and primary fibroblasts, but not in mouse peritoneal macrophages. The inhibitory effect was due to the reduced expression of inducible NO synthase (iNOS), the enzyme responsible for cytokine-induced intracellular NO synthesis, as both agents markedly suppressed the interferon-gamma (IFN-gamma) triggered increase in iNOS concentration in L929 cells. In addition, ADC and paclitaxel prevented the IFN-gamma-triggered activation of p44/p42 mitogen activated protein (MAP) kinase in L929 fibroblasts, suggesting a possible mechanism for the observed inhibition of iNOS expression. These results might have important implications for the therapeutic effect of ADC and paclitaxel, since their inhibitory action on NO release partly neutralized the NO-dependent toxicity of IFN-gamma on L929 fibrosarcoma cells. PMID- 14757127 TI - Quercetin, a flavonoid, inhibits the proliferation, differentiation, and mineralization of osteoblasts in vitro. AB - It is possible that the flavonoids that are found in many foods might have a protective effect against osteoclastic activity. However, little information is available about the effects of flavonoids on osteoblastogenesis. Therefore, we investigated the effects of quercetin, a flavonoid, on the metabolism of rat calvarial osteoblast-like cells (ROB cells) in culture. The proliferation of cells was markedly inhibited upon exposure of cells to quercetin at 5 x 10(-6) to 1 x 10(-5) M. Quercetin at 1 x 10(-5) M did not induce apoptosis in ROB cells but arrested cells at the G1 phase of the cell cycle. In addition, quercetin stimulated the expression of mRNA for p21(waf1/cip1), which inhibits the activity of cyclin-dependent kinases, and inhibited the phosphorylation of histone H1. Furthermore, after cells had ceased to proliferate, quercetin reduced the activity of alkaline phosphatase, the level of expression of mRNA for osteocalcin, the rate of deposition of Ca(2+), and the formation of mineralized nodules, all of which are markers of osteoblast differentiation. These findings indicate that quercetin inhibits the proliferation, differentiation, and mineralization of osteoblastic cells. PMID- 14757128 TI - Allicin (from garlic) induces caspase-mediated apoptosis in cancer cells. AB - Garlic (Allium sativum) has been used for centuries for treating various ailments, and its consumption is said to reduce cancer risk and its extracts and components effectively block experimentally induced tumors. Allicin, the major component present in freshly crushed garlic, is one of the most biologically active compounds of garlic. We found that allicin inhibited the growth of cancer cells of murine and human origin. Allicin induced the formation of apoptotic bodies, nuclear condensation and a typical DNA ladder in cancer cells. Furthermore, activation of caspases-3, -8 and -9 and cleavage of poly(ADP-ribose) polymerase were induced by allicin. The present results demonstrating allicin induced apoptosis of cancer cells are novel since allicin has not been shown to induce apoptosis previously. Our results also provide a mechanistic basis for the antiproliferative effects of allicin and partly account for the chemopreventive action of garlic extracts reported by earlier workers. PMID- 14757129 TI - Wogonin inhibits excitotoxic and oxidative neuronal damage in primary cultured rat cortical cells. AB - The present study evaluated effects of wogonin (5,7-dihydroxy-8-methoxyflavone) on excitotoxic and oxidative stress-induced neuronal damage in primary cultured rat cortical cells. Wogonin was shown to inhibit the excitotoxicity induced by glutamate or N-methyl-D-aspartic acid, whereas it showed no effects on the alpha amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid- or kainate-induced toxicity. In addition, wogonin inhibited the oxidative neuronal damage induced by H(2)O(2), xanthine/xanthine oxidase, and by a glutathione depleting agent D,L-buthionine [S,R]-sulfoximine. Furthermore, wogonin dramatically inhibited lipid peroxidation initiated by Fe(2+) and L-ascorbic acid in rat brain homogenates. It also exhibited 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity. Taken together, these results demonstrate that wogonin exhibits neuroprotective actions in cultured cortical cells by inhibiting excitotoxicity and various types of oxidative stress-induced damage, and that its antioxidant actions with radical scavenging activity may contribute, at least in part, to the neuroprotective effects. PMID- 14757130 TI - Characterisation of Gs activation by dopamine D1 receptors using an antibody capture assay: antagonist properties of clozapine. AB - Herein, we examined the direct coupling of human dopamine D1 receptors to G(s) proteins using an antibody capture assay together with a detection technique employing scintillation proximity assay beads. Using a specific antibody, dopamine (DA) and the selective dopamine D1 receptor agonists, 6-chloro-7,8 dihydroxy-1-phenyl-2,3,4,5-tetrahydro-1H-3-benzazepine (SKF81297) and 3-allyl-6 chloro-7,8-dihydroxy-1-phenyl-2,3,4,5-tetrahydro-1H-3-benzazepine (SKF82958), behaved as high-efficacy agonists ( approximately 100%) in stimulating guanosine 5'-O-(3-[35S]thio)-triphosphate ([35S]GTP gamma S) binding to G(s) in L-cells, whereas 2,3,4,5,-tetrahydro-7,8-dihydroxy-1-phenyl-1H-3-benzazepine (SKF38393) displayed partial agonist properties (70%). The action of dopamine was specifically mediated by human dopamine D1 receptors inasmuch as the selective human dopamine D1 receptor antagonist, (R)-(+)-8-chloro-2,3,4,5-tetrahydro-3 methyl-5-phenyl-1H-benzazepine-7-ol (SCH23390), blocked dopamine-induced [35S]GTP gamma S binding to G(s) with a pK(B) (9.29) close to its pK(i) (9.33). The antipsychotic agents, clozapine and haloperidol, displayed no intrinsic activity when tested alone and inhibited dopamine-stimulated G(s) activation with pK(B)'s of 6.7 and 7.3, respectively, values close to their pK(i) values at these sites. In conclusion, the use of an anti-G(s) protein immunoprecipitation assay coupled to scintillation proximity assays allows direct evaluation of the functional activity of dopamine D1 receptors ligands at the G protein level. Employing this novel technique, the typical and atypical antipsychotics, clozapine and haloperidol, respectively, both exhibited antagonist properties at dopamine D1 receptors. PMID- 14757131 TI - Kappa-opioid receptors are differentially labeled by arylacetamides and benzomorphans. AB - Using Chinese Hamster Ovary cell membranes that stably expressed the human kappa opioid receptor, we investigated the hypothesis that kappa(1)- and kappa(2) opioid receptors, historically defined by their pharmacological selectivity for either arylacetamides or benzomorphans are, in fact, different affinity states or binding sites on the same kappa-opioid receptors. Receptor binding studies showed that GTP gamma S potently inhibited [3H](5 alpha,7 alpha,8 beta)-(+)-N-methyl-N (7-[1-pyrrolidinyl]-1-oxaspiro [4.5]dec-8-yl)-benzeneacetamide (U69,593) binding, compared to virtually no inhibition of [3H]bremazocine binding. Saturation binding experiments showed a three-fold decrease in [3H]U69,593 affinity in the presence of GTP gamma S, but GTP gamma S had no effect on [3H]bremazocine affinity. The kappa-opioid receptor antagonist nor-binaltorphimine had a four fold higher affinity for [3H]U69,593-labeled receptors than for [3H]bremazocine labeled receptors. Functional selectivity studies, measuring the stimulation of [35S]GTP gamma S agonist-induced binding, showed a significantly higher U69,593 induced G protein-receptor activation in comparison to the stimulation observed with bremazocine. These results suggest that pharmacologically defined 1 kappa opioid receptor subtypes may be different affinity states of the same receptor. PMID- 14757132 TI - Ebselen inhibits p38 mitogen-activated protein kinase-mediated endothelial cell death by hydrogen peroxide. AB - Ebselen (2-phenyl-1, 2-benzisoselenazol-3[2H]-one) is a seleno-organic compound exhibiting both glutathione peroxidase and antioxidant activity. Although it has been reported that ebselen is effective against hydrogen peroxide (H(2)O(2)) induced cell death in several cell types, its effect on endothelial cell damage has not yet been elucidated. In the present study, we examined the effect of ebselen on H(2)O(2)-induced human umbilical vein endothelial cells (HUVECs) death, and its intracellular mechanism. Our findings showed that pretreatment of HUVECs with ebselen resulted in a significant recovery from H(2)O(2)-induced cell death in a concentration-dependent manner. In addition to the inhibition of lactate dehydrogenase (LDH) leakage, ebselen inhibited H(2)O(2)-induced cytochrome c release and caspase-3 activation and the resultant apoptosis in HUVECs. Moreover, it was observed that H(2)O(2) significantly stimulated activation of mitogen-activated protein (MAP) kinases, i.e., p38 MAP kinase, c Jun N-terminal kinase (JNK) and extracellular signal-regulated kinase (ERK1/2). Ebselen inhibited H(2)O(2)-induced p38 MAP kinase, but not JNK or ERK1/2 activation. Furthermore, SB203580 (4-[4-fluorophenyl]-2-[4-methylsulfinylphenyl] 5-[4-pyridyl]-1H-imidazole), a specific p38 MAP kinase inhibitor, inhibited H(2)O(2)-induced p38 MAP kinase phosphorylation, cytochrome c release, caspase-3 activation, as well as cell death in HUVECs. These findings suggest that ebselen attenuates H(2)O(2)-induced endothelial cell death through the inhibition of signaling pathways mediated by p38 MAP kinase, caspase-3, and cytochrome c release. Thus, inhibition of p38 MAP kinase by ebselen may imply its usefulness for prevention and/or treatment of endothelial cell dysfunction, which was suggested to be the first step in the development of atherosclerosis. PMID- 14757133 TI - Effect of the ecto-ATPase inhibitor, ARL 67156, on the bovine chromaffin cell response to ATP. AB - Bovine chromaffin cells contain an ecto-ATPase (K(m)=1.57 +/- 0.27 x 10(-4) M) which can hydrolyze ATP present in the culture media. ARL 67156 is a competitive inhibitor of this ATPase (K(i)=2.55 +/- 1.36 x 10(-7) M). A small increase in potency (threefold) is seen when ARL 67156 is included during measurement of ATP stimulated inositol phosphate formation. ARL 67156 also acts on chromaffin cell P2Y receptors to increase inositol phosphate formation (EC(50)=4.9 x 10(-5) M). It is useful as an ecto-ATPase inhibitor in studies with bovine chromaffin cells since it exhibits a 300-fold selectivity for the ecto-ATPase versus the P2Y receptor. PMID- 14757134 TI - Protein kinase C-beta inhibition and diabetic microangiopathy: effects on endothelial permeability responses in vitro. AB - Protein kinase C (PKC)-beta and other PKC isozymes have been implicated in the loss of endothelial barrier function in diabetic microangiopathy. The effects of a PKC-beta-specific inhibitor, LY379196, on hyperpermeability responses to high glucose, angiotensin II, alpha-thrombin and endothelin-1 were evaluated using an in vitro model of human pulmonary artery endothelial cell monolayers. LY379196 attenuated the increase in transendothelial albumin flux induced by glucose 40 mM (e.g. 411+/-160% [high-glucose] vs. 167+37% [high-glucose+LY379196], P<0.001) and angiotensin II 10 microM (e.g. 121+/-12% vs. 246+/-35%, P<0.01); endothelin-1 had no significant effect on monolayer permeability. LY379196 had no significant effect on the marked hyperpermeability response to alpha-thrombin 1 microM. Thus, two major pathways involved in vascular leakage in diabetic microangiopathy are amenable to therapeutic blockade by PKC-beta inhibition. PMID- 14757135 TI - Anxiolytic- and antidepressant-like profile of a new CRF1 receptor antagonist, R278995/CRA0450. AB - 1-[8-(2,4-dichlorophenyl)-2-methylquinolin-4-yl]-1,2,3,6-tetrahydropyridine-4 carboxamide benzenesulfonate (R278995/CRA0450) is a newly synthesized corticotropin-releasing factor subtype 1 (CRF(1)) receptor antagonist. In the present study, in vitro and in vivo pharmacological profiles of R278995/CRA0450 were investigated. R278995/CRA0450 showed high affinity for recombinant and native CRF(1) receptors without having affinity for the CRF(2) receptor. R278995/CRA0450 attenuated CRF-induced cyclic AMP formation in AtT-20 cells and CRF-induced forepaw treading in gerbils, indicating that R278995/CRA0450 is an antagonist of the CRF(1) receptor. In addition to CRF(1) receptor antagonism, R278995/CRA0450 showed high affinity for the sigma(1) receptor, and attenuated (+)-SKF10,047-induced head-weaving behavior, suggesting sigma(1) receptor antagonism. R278995/CRA0450 showed dose-dependent in vivo occupancy when assessed by ex vivo receptor binding, indicating good brain penetration. R278995/CRA0450 did not alter spontaneous anxiety when tested in the rat elevated plus maze (up to 3 mg/kg, p.o.) or lick suppression test (up to 10 mg/kg, i.p.). However, potent anxiolytic-like properties were observed in rats subjected to swim stress prior to testing on the elevated plus-maze, indicating activity primarily in tests taxing stress-induced anxiety. R278995/CRA0450 was inactive in mouse tail suspension, rat forced swim and rat differential-reinforcement-of-low-rate 72-s (DRL72), while it showed dose-dependent antidepressant-like effects in the rat learned helplessness paradigm and the olfactory bulbectomy model, demonstrating activity in a subset of animal models of depression associated with subchronic stress exposure. No or only mild effects were seen in tests of locomotor activity, motor coordination and sedation. These results indicate that R278995/CRA0450 is an orally active CRF(1) and sigma(1) receptor antagonist with potent anxiolytic-like and antidepressant-like activities. PMID- 14757136 TI - Levetiracetam improves choreic levodopa-induced dyskinesia in the MPTP-treated macaque. AB - L-3,4 dihydroxyphenylalanine (levodopa)-induced dyskinesia in Parkinson's disease patients is characterized by a mixture of chorea and dystonia. Electrophysiological studies suggest that chorea is associated with abnormal synchronization of firing of basal ganglia neurons while dystonia is not. Levetiracetam is a novel anti-epileptic drug known to exhibit unique desynchronizing properties in contrast to other anti-epileptic drugs. We assessed the anti-dyskinetic efficacy of levetiracetam (13, 30 and 60 mg/kg, p.o.) administered in combination with an individually tailored dose of levodopa (Levodopa/carbidopa, 4:1 ratio, 19+/-1.8 mg/kg, p.o.), in six dyskinetic 1-methyl 4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-lesioned macaques. Levetiracetam (60 mg/kg) significantly reduced levodopa-induced chorea during the first hour post treatment but had no effect on dystonia. Levetiracetam, at all doses tested, had no effect on the anti-parkinsonian action of levodopa. These results suggest that levetiracetam may provide a novel therapeutic approach specifically aimed at the choreic form of levodopa-induced dyskinesia. PMID- 14757137 TI - Profile of spinal and supra-spinal antinociception of (-)-linalool. AB - We previously reported that administration of (-)-linalool, the naturally occurring enantiomer in essential oils, induced a significant reduction in carrageenin-induced oedema and in acetic acid-induced writhing. The latter effect was completely antagonised by the muscarinic receptor antagonist atropine and by the opioid receptor antagonist naloxone. To further characterise the antinociceptive profile of (-)-linalool, we studied its effect in the hot plate and the formalin in tests. In addition, to determine the possible involvement of the cholinergic, opioidergic and dopaminergic systems, we tested the effects of atropine, pirenzepine, a muscarinic M1 receptor antagonist, naloxone, sulpiride, a dopamine D2 receptor antagonist and (R)-(+)-7-chloro-8-hydroxy-3-methyl-1 phenyl-2,3,4,5-tetrahydro-1H-3-benzazepine (SCH-23390), a dopamine D1 receptor antagonist on (-)-linalool-induced antinociception. Moreover, since K(+) channels seem to play an important role in the mechanisms of pain modulation, we examined the effect of glibenclamide, an ATP-sensitive K(+) channel inhibitor on (-) linalool-induced antinociception. The administration of (-)-linalool (100 and 150 mg/kg, s.c.) increased the reaction time in the hot-plate test. Moreover, (-) linalool (50 and 100 mg/kg) produced a significant reduction in the early acute phase of the formalin model, but not in the late tonic phase. The highest dose (150 mg/kg) caused a significant antinociceptive effect on both phases. The antinociceptive effects of (-)-linalool were decreased by pre-treatment with atropine, naloxone, sulpiride and glibenclamide but not by pirenzepine and SCH 23390. These results are in agreement with the demonstrated pharmacological properties of linalool, mainly its cholinergic, local anaesthetic activity and its ability to block NMDA receptors. Furthermore, a key role seems to be played by K(+) channels, whose opening might be the consequence of a stimulation of muscarinic M2, opioid or dopamine D2 receptors. PMID- 14757138 TI - Catalepsy induced by intra-striatal administration of nitric oxide synthase inhibitors in rats. AB - Systemic administration of nitric oxide synthase (NOS) inhibitors induces catalepsy in a dose-dependent manner in male Albino-Swiss mice. The objective of the present work was to investigate if similar effects occur in rats and if these effects are centrally mediated. The results showed that systemic administration of N(G)-nitro-L-arginine (L-NOARG, 40-160 mg/kg, i.p.), a non-selective NOS inhibitor, induced catalepsy in rats. Similar effects were found after intracerebroventricular (i.c.v.) injection of L-NOARG (50-200 nmol) or N(G)-nitro L-arginine methylester (L-NAME, 100-200 nmol). The dose-response curve of the former compound, however, had an inverted U shape. The effect of L-NOARG (100 nmol, i.c.v.) was completely prevented by pre-treatment with L-arginine (300 nmol, i.c.v.) but not by D-arginine (300 nmol, i.c.v.). Intra-striatal injection of N(G)-monomethyl-L-arginine (L-NMMA, 100 nmol), 7-nitroindazole (7-NIO, 100 nmol), L-NOARG (25-100 nmol) or L-NAME (50-200 nmol) also induced catalepsy. Similar to i.c.v. administration, the latter two compounds produced bell-shaped dose-response curves. The cataleptic effect of intra-striatal administration of L NAME (100 nmol) was reversed by local treatment with L-arginine (100 nmol). These results suggest that interference with the striatal formation of nitric oxide may induce significant motor effects in rats. PMID- 14757139 TI - Effect of late treatment with gamma-hydroxybutyrate on the histological and behavioral consequences of transient brain ischemia in the rat. AB - It has been previously described that gamma-hydroxybutyrate (GHB) provides significant protection against transient global cerebral ischemia in the rat (four vessel occlusion model), when given 30 min before or 10 min after artery occlusion. Here, we show that in the same rat model, significant protection can also be obtained when treatment is started 2 h after the ischemic episode. In saline-treated animals, 30 min of global ischemia followed by reperfusion caused a massive loss of neurons in the hippocampal CA1 subfield (examined 63 days after the ischemic episode), and an impairment of sensory-motor performance (tested on the 51st and 63rd days after ischemia) and of spatial learning and memory (evaluated starting 46 days after the ischemic episode). Treatment with GHB--300 mg/kg intraperitoneally (i.p.) 2 h after the ischemia-reperfusion episode, followed by 100 mg/kg i.p. twice daily for the following 10 days--afforded a highly significant protection, against both histological damage and sensory-motor and learning-memory impairments. These data further suggest the possible therapeutic effectiveness of GHB in brain ischemia, and indicate that the underlying mechanism of action involves non-immediate steps of the ischemia induced cascade of events. PMID- 14757140 TI - Loss of amitriptyline analgesia in alpha 2A-adrenoceptor deficient mice. AB - Tricyclic antidepressants have analgesic and sedative effects in addition to their antidepressive properties. We tested the acute analgesic and locomotor inhibitory effects of the tricyclic antidepressant amitriptyline and the alpha(2) adrenoceptor agonist clonidine in wild-type control and in alpha(2A)-adrenoceptor knockout mice in hot-plate and tail-flick tests. Amitriptyline-induced analgesia was lost in alpha(2A)-adrenoceptor knockout mice. The locomotor inhibitory effect of amitriptyline was reduced, but not fully abolished in alpha(2A)-adrenoceptor knockout mice. Similar results were obtained with clonidine. We conclude that alpha(2A)-adrenoceptors appear to have a significant role in amitriptyline induced acute analgesia in mice, and that alpha(2A)-adrenoceptors also participate in the sedative effects of amitriptyline. PMID- 14757141 TI - Tonic inhibition by orphanin FQ/nociceptin of noradrenaline neurotransmission in the amygdala. AB - The present microdialysis study investigated whether nociceptin/orphanin FQ exerts a tonic inhibition of the release of noradrenaline in the basolateral nucleus of the amygdala in awake rats. The non-peptide competitive nociceptin/orphanin FQ (N/OFQ) peptide receptor antagonist J-113397 (20 mg/kg i.p.) induced an increase in the release of noradrenaline to about 150-200%. The increase was strongly suppressed by local infusion of an endogenous N/OFQ peptide receptor agonist, nociceptin/orphanin FQ (1 microM) via retrograde microdialysis, into the basolateral nucleus of the amygdala. Local infusion of nociceptin/orphanin FQ (1 microM) itself reduced noradrenaline release in the basolateral nucleus of the amygdala to about 70% of basal levels. These results indicate that a large part of basal release of noradrenaline in the basolateral nucleus of the amygdala is under tonic inhibitory control by endogenous nociceptin/orphanin FQ through the N/OFQ peptide receptors localized within the basolateral nucleus of the amygdala. PMID- 14757142 TI - Facilitation of morphine withdrawal symptoms and morphine-induced conditioned place preference by a glutamate transporter inhibitor DL-threo-beta benzyloxyaspartate in rats. AB - There is a body of evidence implying the involvement of the central glutamatergic system in morphine dependence. In this study, we examined the effect of intracerebroventricular (i.c.v.) administration of a potent glutamate transporter inhibitor, DL-threo-beta-benzyloxyaspartate (DL-TBOA), on acute morphine-induced antinociception, expression of somatic and negative affective components of morphine withdrawal, and acquisition of morphine-induced conditioned place preference in rats. I.c.v administration of DL-TBOA (10 nmol) to naive rats did not affect the acute antinociceptive effect of morphine. I.c.v. administration of DL-TBOA (10 nmol) to morphine-dependent rats significantly facilitated the expression of naloxone-precipitated somatic signs and conditioned place aversion. DL-TBOA (3 and 10 nmol) significantly facilitated acquisition of morphine-induced conditioned place preference. DL-TBOA itself produced neither conditioned place aversion nor place preference in naive rats. These results suggest that central glutamate transporters play inhibitory roles in the expression of somatic and negative affective components of morphine withdrawal and the reinforcing effect of morphine. PMID- 14757143 TI - Intravenous self-administration of abused solvents and anesthetics in mice. AB - Volatile organic solvents, fuels and anesthetics are subject to abuse. The aim of the present study was to evaluate i.v. self-administration of several of these chemicals in drug- and experiment-naive mice using a commercially available vehicle, intralipid. Two strains of mice (DBA/2 and Swiss) were allowed to self administer toluene (0.0017-0.17 micromol/infusion), 1,1,1-trichloroethane (0.006 0.19 micromol/infusion), ethanol (0.32-1.6 micromol/infusion), cyclohexane (0.0017-0.052 micromol/infusion), propofol (0.01-0.53 micromol/infusion) and flurothyl (0.00042-0.072 micromol/infusion) or their vehicles during 30-min tests. During the test, each nose-poke of the master mouse resulted in a 1.88 microl i.v. infusion to the master mouse and a yoked control mouse. When the delivery line was loaded with a reinforcing drug solution, the number of nose pokes of the master mice significantly exceeded that for yoked control mice. In the present experiments, significant differences in rates of nose-poking were observed between mice receiving response-contingent and response-noncontingent deliveries of ethanol and toluene in both strains of mice and of 1,1,1 trichloroethane in Swiss mice. These data suggest that the reinforcing effects of abused inhalants can be studied using i.v. self-administration procedures. PMID- 14757144 TI - Mechanisms underlying endothelium-dependent flow increase in perfused rat mesenteric vascular bed. AB - The isolated rat mesenteric vasculature was perfused at constant pressures of 40, 80 or 120 mm Hg and the change in flow rate was measured. In the presence of phenylephrine, treatment with 3-[(3-cholamidopropyl) dimethylammonio]-1-propane sulfonate (CHAPS) or N(G)-nitro-L-arginine (L-NA) significantly inhibited the pressure-dependent flow rate increase, but treatment with indomethacin or charybdotoxin plus apamin did not. Acetylcholine, bradykinin and ADP increased the flow rate, which had been markedly suppressed by CHAPS. At 80 mm Hg, the flow rate increase induced by these agonists was not affected by indomethacin plus L NA, but was suppressed by subsequent treatment with charybdotoxin plus apamin. Changes in the perfusion pressure did not significantly affect the flow rate increases induced by the agonists. In conclusion, the opening of charybdotoxin plus apamin-sensitive Ca(2+)-dependent K(+) channels may be mainly involved in the endothelium-dependent flow rate increase induced by the agonists, whereas nitric oxide (NO) may be responsible for the endothelium-dependent, pressure induced flow rate increase. PMID- 14757145 TI - Blockade of beta 1- and desensitization of beta 2-adrenoceptors reduce isoprenaline-induced cardiac fibrosis. AB - The aim of the present study was to analyse the role of beta(1)- and beta(2) adrenoceptors in the catecholamine-induced myocardial remodeling, especially the interstitial fibrosis. Wistar rats were subjected to a 2-week chronic isoprenaline administration (30 microg/kg/h). Rats received a concomitant treatment with the selective beta(1)-adrenoceptor antagonist, bisoprolol (50 mg/kg/day p.o.) or were chronically pretreated with the selective beta(2) adrenoceptor agonist salbutamol (40 microg/kg/h) for 1 week to induce beta(2) adrenoceptor desensitization. The pretreatment with salbutamol induced a 59% down regulation of left ventricular beta(2)-adrenoceptors compared to control. The extent of the isoprenaline-induced left ventricular fibrosis was significantly reduced in both the bisoprolol and salbutamol groups compared with the control isoprenaline-treated group especially in the apical region (1.7+/-0.6% and 1.4+/ 0.3% versus 6.0+/-1.3%, respectively, P<0.005). beta(1)-adrenoceptor blockade and beta(2)-adrenoceptors down-regulation provided similar protection against isoprenaline-induced cardiac interstitial fibrosis suggesting that both beta adrenoceptors are involved in such cardiac remodeling process. PMID- 14757146 TI - Vasopeptidase inhibition reverses myocardial vasoactive intestinal peptide depletion and decreases fibrosis in salt sensitive hypertension. AB - We have shown previously that the concentration of Vasoactive Intestinal Peptide (VIP) in the heart is inversely correlated with the degree of fibrosis in a number of experimental models of early myocardial fibrosis. Vasopeptidase inhibition and angiotensin converting enzyme inhibition both decrease myocardial fibrosis. In this study, we sought to determine whether this myocardial protective effect might reflect increased VIP concentrations in the heart. We compared the effects of 4 weeks treatment of the vasopeptidase inhibitor omapatrilat and the angiotensin converting enzyme inhibitor enalapril on the degree of fibrosis and the concentration of VIP in the heart in salt sensitive hypertension induced by treatment with L-nitro-omega-methylarginine (L-NAME). Systolic blood pressure decreased in both treatment groups compared with control (omapatrilat P<0.005; enalapril P<0.001). Myocardial fibrosis was less for omapatrilat than control (P<0.0005) and enalapril (P<0.0005) groups. Myocardial VIP was greater in omapatrilat than in controls (P<0.005) and enalapril-treated rats (P<0.05). We conclude that vasopeptidase inhibition exerts a greater myocardial protective effect than angiotensin converting enzyme inhibition. Further, this myocardial protective effect is associated with increased VIP in the heart suggesting a pathogenetic role for VIP depletion in the development of fibrosis in the heart. PMID- 14757147 TI - Angiotensin II stimulates superoxide production via both angiotensin AT1A and AT1B receptors in mouse aorta and heart. AB - The present study was conducted to determine the roles of angiotensin AT(1A) and AT(1B) receptors in angiotensin II-induced superoxide anion production in mouse aorta and heart. Superoxide anion production in aorta was determined by the lucigenin chemiluminescence method, and thiobarbituric acid reactive substances in heart tissues were measured by biochemical assay. The basal production rate of superoxide anion in aorta of wild type (WT) mice was significantly higher than in angiotensin AT(1A) receptor knockout (AT(1A) KO) mice. Angiotensin II (2.8 mg/kg/day, s.c. for 13 days) significantly increased superoxide anion production in aorta of both AT(1A) KO and WT mice. However, the superoxide anion production rate in aorta of angiotensin II-infused AT(1A) KO mice was significantly lower than in angiotensin II-infused WT mice. Valsartan (40 mg/kg/day in drinking water) prevented angiotensin II-induced superoxide anion production in aorta of WT and AT(1A) KO mice. Similarly, thiobarbituric acid reactive substances levels in heart tissues of angiotensin II-treated WT and AT(1A) KO mice were significantly higher than those in vehicle-infused WT and AT(1A) KO mice, respectively. Valsartan prevented angiotensin II-induced increases of thiobarbituric acid reactive substances levels in heart tissue of both WT and AT(1A) KO mice. These results indicate that angiotensin II stimulates superoxide anion production via both angiotensin AT(1A) and AT(1B) receptors, and that angiotensin AT(1A) receptors appear to play a predominant role in angiotensin II induced superoxide anion production in mouse aorta and heart. PMID- 14757148 TI - Chronic ethanol administration attenuates imidazoline I1 receptor- or alpha 2 adrenoceptor-mediated reductions in blood pressure and hemodynamic variability in hypertensive rats. AB - Our previous studies have demonstrated that acute ethanol administration counteracts imidazoline I(1) receptor but not alpha(2)-adrenoceptor-mediated hypotension in spontaneously hypertensive rats (SHR). In the present study, we investigated the effect of chronic ethanol administration on hypotensive responses elicited by acute administration of selective imidazoline I(1) receptor (rilmenidine) or alpha(2)-adrenoceptor (alpha-methyldopa) agonist along with ethanol effects on: (i) locomotor activity and (ii) time-domain indices of variability in blood pressure (standard deviation of mean arterial pressure) and heart rate (standard deviation of beat-to-beat intervals and root mean square of successive differences in R-R intervals). Hemodynamic and locomotor responses elicited by rilmenidine or alpha-methyldopa were assessed in radiotelemetered ethanol-fed (2.5% or 5% w/v, 12 week) and control SHR. In control SHR, i.p. rilmenidine (600 microg/kg) or alpha-methyldopa (100 mg/kg) significantly reduced blood pressure. Rilmenidine had no effect on heart rate whereas alpha-methyldopa elicited a biphasic response (tachycardia followed by bradycardia). Blood pressure and heart rate oscillations were also reduced by both drugs, which may conform to sympathoinhibition. The hypotensive effect of rilmenidine or alpha methyldopa was significantly attenuated by ethanol feeding (2.5% or 5%) in a concentration-dependent manner. In addition, ethanol attenuated alpha-methyldopa evoked reduction in heart rate, but not blood pressure, variability in marked contrast to attenuating rilmenidine-evoked reductions in blood pressure, but not heart rate, variability. These findings demonstrate that, unlike its acute effects, chronic ethanol attenuates both imidazoline I(1) receptor and alpha(2) adrenoceptor-mediated hypotension whereas its effect on hemodynamic variability depended on the nature of the hypotensive stimulus. PMID- 14757149 TI - Rho-kinase expression and its contribution to the control of perfusion pressure in the isolated rat mesenteric vascular bed. AB - Rho-kinase expression was investigated in the rat mesenteric artery and the effects of its inhibitors, (+)-(R)-trans-4-(1-aminoethyl)-N-(4-pyridyl) cyclohexanecarboxamide dihydrochloride monohydrate (Y-27632) and fasudil (HA 1077), were examined on the increase in perfusion pressure induced by two different receptor agonists, namely the alpha-adrenoceptor agonist, phenylephrine and, the endothelin ET(A) and ET(B) receptor agonist, endothelin-1. Y-27632 and fasudil produced a concentration-dependent decrease in perfusion pressure. There was no difference between the concentration-response lines of these two inhibitors. The maximum decrease in the perfusion pressure induced by 10(-5) M Y 27632 was 85.8+/-3.7% when the tone was increased by phenylephrine. However, it was 48.1+/-5.4% (P<0.001) when the perfusion pressure was elevated by endothelin 1. Saponin perfusion (100 mg l(-1), for 10 min), which abolished acetylcholine induced relaxation, did not significantly modify the Y-27632-elicited relaxation. Western blot analysis revealed that rat mesenteric artery expresses Rho-kinase protein with a molecular weight of approximately 160 kDa. These results show that Rho-kinase enzyme is expressed in rat mesenteric artery and that it contributes to the control of vascular resistance. Moreover, endothelium removal had no marked effect on the vasodilatation induced by Y-27632. In addition, the endothelin-1-induced vasoconstriction was more resistant to the Rho-kinase inhibitors than was that induced by phenylephrine, probably because excitatory endothelin receptors are associated with this signal transduction pathway at a different level from that of alpha-adrenoceptors. PMID- 14757150 TI - Neocuproine inhibits the decomposition of endogenous S-nitrosothiol by ultraviolet irradiation in the mouse gastric fundus. AB - In the present study, we investigated whether copper ions are involved in the decomposition of endogenous S-nitrosothiols by ultraviolet (UV) light irradiation in the mouse gastric fundus. The effects of copper ions and chelators of copper(I) and copper(II), neocuproine and cuprozine, respectively, were studied on relaxations in response to S-nitrosoglutathione, UV irradiation, exogenous nitric oxide (NO), added as acidified NaNO(2), and isoproterenol. UV irradiation of smooth muscle strips induced fast and transient relaxations which were mimicked by exogenous NO. S-Nitrosoglutathione induced concentration-dependent relaxations, which were more sustained than those elicited by UV irradiation or NO. CuCl(2) did not affect relaxations elicited by UV irradiation, exogenous NO and isoproterenol but enhanced those elicited by S-nitrosoglutathione. CuSO(4) but not FeSO(4) mimicked the effect of CuCl(2) on relaxations elicited by S nitrosoglutathione. Neocuproine, the copper(I)-specific chelator, inhibited both photorelaxation and S-nitrosoglutathione-induced relaxation, and this inhibition was prevented by CuCl(2). In contrast, neocuproine significantly enhanced the relaxations in response to exogenous NO, without affecting the relaxations elicited by isoproterenol. Cuprizone, a specific copper(II) chelator, did not affect relaxations in response to S-nitrosoglutathione, UV irradiation, exogenous NO and isoproterenol. These results suggest that copper(I) and not copper(II) may play a role in the NO release evoked by the light-induced decomposition of endogenous S-nitrosothiols in mouse gastric fundus. Also, results with the selective copper(I) chelator, neocuproine, confirmed our recent findings that the endogenous "store" of S-nitrosoglutathione, rather than NO, acts as an intermediate in photorelaxation of the mouse gastric fundus, and that photorelaxation may be a suitable model to elucidate the nature of endogenous S nitrosothiols. PMID- 14757151 TI - Dexamethasone delays ulcer healing by inhibition of angiogenesis in rat stomachs. AB - Using the non-ulcerogenic doses of dexamethasone, we explored the action of glucocorticoids on ulcer healing and its relationship with angiogenic factors in the gastric mucosa. We applied dexamethasone (0.1 or 0.2 mg/kg/day) intragastrically in rats with acetic acid-induced gastric ulcer. The mucosal prostaglandin E(2) level and protein expressions of basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF) at the ulcer margin were determined. Ulcer induction significantly increased protein expressions of bFGF, VEGF, and prostaglandin E(2) level at the ulcer margin together with angiogenesis at the ulcer margin and base. The non-ulcerogenic doses of dexamethasone inhibited angiogenesis at the ulcer margin and ulcer base and delayed ulcer healing. These were associated with a significant decrease of prostaglandin E(2) level and VEGF expression, but not the bFGF expression. Supplementation with prostaglandin E(2) attenuated the inhibitory action of dexamethasone on VEGF expression and reversed the adverse effects of dexamethasone on angiogenesis and ulcer healing, without influencing bFGF expression. We concluded that dexamethasone given at non-ulcerogenic doses could decrease angiogenesis and delay acetic acid-induced ulcer healing; these actions were at least, in part, due to depletion of prostaglandin E(2) level followed by down-regulation of VEGF at the ulcer margin of the stomach. PMID- 14757152 TI - T-0156, a novel phosphodiesterase type 5 inhibitor, and sildenafil have different pharmacological effects on penile tumescence and electroretinogram in dogs. AB - T-0156 (2-(2-methylpyridin-4-yl)methyl-4-(3,4,5-trimethoxyphenyl)-8-(pyrimidin-2 yl)methoxy-1,2-dihydro-1-oxo-2,7-naphthyridine-3-carboxylic acid methyl ester hydrochloride) is a newly synthesized phosphodiesterase type 5 inhibitor, and its potency and selectivity are higher than those of sildenafil in an enzyme assay. In the present study with anesthetized dogs, we examined the effects of intravenous T-0156 or sildenafil on the pelvic nerve stimulation-induced penile tumescence and light-adapted flicker stimulation-induced electroretinogram, parameters of which are reported to be indicators for inhibition of phosphodiesterase type 5 and type 6, respectively. Both compounds potentiated the penile tumescence in a dose-dependent manner. T-0156 at 10 microg/kg and sildenafil at 100 microg/kg showed almost the same potentiating percentage (181.5+/-31.1% and 190.0+/-37.9%) in spite of the plasma concentration of T-0156 being about five times lower than that of sildenafil (16.7+/-1.6 and 78.8+/-5.3 ng/ml), indicating that the effect of T-0156 on tumescence is more potent than that of sildenafil. While the high dose of T-0156 (1000 microg/kg) reduced the amplitude and increased the latency of the electroretinogram positive wave, the effects of T-0156 were conversely weaker than those of sildenafil (reduction of amplitude; T-0156: 41.1+/-8.0%, sildenafil: 71.7+/-3.9%, increase of latency; T 0156: 3.9+/-0.6%, sildenafil: 14.5+/-1.4%, at 1000 microg/kg). These results clearly showed the difference in the properties of T-0156 and sildenafil in pharmacological studies with anesthetized dogs, and the difference appeared to correspond with their inhibitory potencies for phosphodiesterase type 5 and type 6. It was concluded that T-0156 would be a useful pharmacological tool as a potent and highly selective phosphodiesterase type 5 inhibitor. PMID- 14757153 TI - Comparison of effects of cyclooxygenase inhibitors on myometrial contraction and constriction of ductus arteriosus in rats. AB - The aim of this study was to compare the tocolytic effect of a selective cyclooxygenase-2 inhibitor, DFU (5,5-dimethyl-3(3-fluorophenyl)-4-(4 methylsulphonyl)phenyl-2(5H)-furanone), indomethacin and nimesulide on myometrial strips isolated from rats in both lipopolysaccharide-induced preterm labour and term labour. We also compared the constrictor effects of DFU and indomethacin on the fetal ductus arteriosus. Myometrial strips were obtained from preterm and term labour Wistar albino rats and were mounted in organ baths for the recording of isometric tension. DFU, nimesulide and indomethacin significantly inhibited KCl-, oxytocin-, prostaglandin E(2)- and prostaglandin F(2 alpha)-stimulated contractions of myometrial strips isolated from rats in preterm and term labour. The E(max) value of indomethacin was significantly lower than those for DFU and nimesulide (P<0.05), with no change-log (10) EC(50) values. There was no significant difference between in -log (10) EC(50) and E(max) values of DFU and nimesulide for any of the tissues (P>0.05). In addition, there was no significant difference between -log (10) EC(50) and E(max) values for each of these three agents in myometrial tissues isolated from rats in preterm and term labour (P>0.05). Fetal ductus arteriosus was significantly constricted by DFU (10 or 100 mg/kg) in preterm and term rats, although DFU (10 or 100 mg/kg)-induced constriction ratios were significantly lower than those for indomethacin (P<0.05). These data demonstrate that DFU, a specific cyclooxygenase-2 inhibitor, could be considered as a new therapeutic agent for preterm labour. However, careful attention should be given to constriction of the fetal ductus arteriosus. PMID- 14757154 TI - Effects of TAK-802, a novel acetylcholinesterase inhibitor, on distension-induced rhythmic bladder contractions in rats and guinea pigs. AB - In the present study, we investigated the effects of 8-[3-[1-[(3 fluorophenyl)methyl]-4-piperidinyl]-1-oxopropyl]-1,2,5,6-tetrahydro-4H pyrrolo[3,2,1-ij]quinolin-4-one (TAK-802), a novel acetylcholinesterase inhibitor, on distension-induced rhythmic bladder contractions in urethane anesthetized rats and guinea pigs. TAK-802 potently inhibited human-erythrocyte derived acetylcholinesterase activity with an IC(50) value of 1.5 nM, which represented a potency 30 and 250 times greater than that of the two carbamate acetylcholinesterase inhibitors, neostigimine and distigmine, respectively. Unlike the carbamate acetylcholinesterase inhibitors, TAK-802 exhibits high selectivity for acetylcholinesterase inhibition over butyrylcholinesterase inhibition. In an assay conducted to measure the muscarinic and nicotinic actions, TAK-802 was found to exhibit higher selectivity for muscarinic actions over nicotinic actions in comparison to distigmine. Both TAK-802 and distigmine increased isovolumetric bladder contractions in rats and guinea pigs in a dose dependent manner, with a minimum effective dose (MED) of 0.01 and 0.03 mg/kg i.v., respectively, in rats, and 0.01 and 0.1 mg/kg i.v., respectively, in guinea pigs. The effects of both the drugs were completely abolished by atropine. These results suggest that TAK-802 and other acetylcholinesterase inhibitors can effectively increase reflex bladder contractions by increasing the efficacy of acetylcholine released by nerve impulses. On the other hand, bethanechol, a muscarinic agonist, markedly changed the pattern of distension-induced bladder contractions when administered at the dose of 1 mg/kg i.v., and it did not necessarily augment well-coordinated bladder contractions. Thus, considering that it has some selectivity for muscarinic action, TAK-802 might be expected to be useful in the treatment of voiding dysfunction caused by impaired detrusor contractility. PMID- 14757155 TI - Alpha-helical structure in the C-terminus of vasoactive intestinal peptide: functional and structural consequences. AB - The conformational properties of vasoactive intestinal peptide (VIP) include the N-terminal randomized structure and the C-terminal long alpha-helical structure. We have previously observed that the N-terminal random coil structure plays a crucial role in the receptor-selectivity. Here, to clarify how the formation of the alpha-helix plays a role in its biological functions, we chemically synthesized VIP analogues modified at the C-terminus, mid-chain, and N-terminus of the alpha-helical region, and evaluated the relationship between their alpha helical contents and their biological activities including relaxant effects on murine stomach and receptor-binding activities. VIP and VIP-(1-27) showed equipotent biological activities with 48% and 50% alpha-helical content, respectively, each of which corresponds to 14 amino acid residues. VIP-(1-26) was 10% and threefold less potent in relaxant and binding activities, respectively, compared with VIP, and its 49% alpha-helical content resulted in 13 residues involved in the alpha-helix. Further truncation from 25 to 21 resulted in decrease in the alpha-helical content from 43% to 29%, corresponding residues from 11 to 6, the relaxant activity from 72% to 4%, and the affinity to the membrane from 60-fold to over 10(4)-fold less potency. In addition, disruption of the mid-chain and the N-terminus in the alpha-helical stretch by oxidation of Met(17) and deletion of Thr(11) also inhibited biological activities. These findings suggest that the presence of alpha-helical structure forming in 14 amino acid residues between position 10 and 23 in VIP is essential to its biological functions and the C-terminal amino acid residues between position 24 and 27 are requisite for this alpha-helical formation. PMID- 14757156 TI - Thromboxane A2 (TP) receptor in the non-pregnant porcine myometrium and its role in regulation of spontaneous contractile activity. AB - Although there are species-related differences in uterine prostanoid receptor subtypes, functional prostanoid receptors in the porcine uterus are similar with those in the human uterus (FP, TP, EP(1), EP(2), EP(3), DP and IP) except for the TP receptor. These similarities promoted us to determine whether TP receptors are present in the non-pregnant porcine uterus. For this purpose, the effects of TP receptor agonists and antagonists were investigated by a contraction study and by a binding study. 9,11-Dideoxy-9 alpha, 11 alpha-methanoepoxy-prosta-5Z,13E-dien-1 oic acid (U46619, 1 nM-10 microM), a stable thromboxane A(2) mimetic, caused tetrodotoxin-resistant contraction in both longitudinal and circular muscles of the uterine cornu. The pEC(50) value in the longitudinal muscle (6.69) was lower than that in the circular muscle (7.62), but the maximum response in the longitudinal muscle was two times larger than that in the circular muscle. The longitudinal and circular muscles of other regions (corpus and cervix) also responded to U46619, and region-related difference in contractile responses was observed only in the longitudinal muscles. 4(Z)-6-(2-o-Chlorophenyl-4-o hydroxyphenyl-1,3-dioxan-cis-5-yl) hexenoic acid (ICI192605) and 7-[3-[[2 [(phenylamino)carbonyl] hydrazino]methyl]7-oxabicyclo[2.2.1]hept-2-yl]-,[1S-[1 alpha,2 alpha(Z),3 alpha,4 alpha]]-]5-heptenoic acid (SQ29548) inhibited the contractile responses to U46619 competitively. The longitudinal and circular muscles in the cornu contained a single class of [3H]SQ29548 binding site with similar K(d) values (30 nM), but B(max) in the circular muscle (90.9+/-8.6 fmol/mg protein) was two times higher than that in the longitudinal muscle (58.2+/-8.6 fmol/mg protein). The ranking order of competition by TP receptor agonists and antagonists (with pK(i) values in parentheses) was [1S [1,2(Z),3(1E,3S*),4]]-7-[3-[3-Hydroxy-4-(4-iodophenoxy)-1-butenyl]-7 oxabicyclo[2.2.1]hept-2-yl]-5-heptenoic acid (I-BOP, 7.70)>SQ29548 (7.39)>7-[3-(3 Hydroxy-1-octenyl)bicycle[3.1.1]hept-2-yl]-,[2S-[2 alpha(Z),3 beta(1E,3R*)]]-5 heptenoic acid (CTA(2), 6.55)>7-[3-(3-hydroxy-1-octenyl)-6,6 dimethylbicyclo[3.1.1]hept-2-yl-,[1S-[1 alpha,2 beta(Z),3 alpha(1E,3R*),5 alpha]] 5-heptenoic acid (PTA(2), 6.50)>U46619 (6.41)>7-[5-(3-hydroxy-1-octenyl)-2 oxabicyclo[2.2.1] hept-6yl]-,[1S-[1 alpha,4 alpha,5 alpha(1E,3R*),6 beta(Z)]]-5 heptenoic acid (U44069, 6.34), and this order is consistent with current TP receptors. Treatment with indomethacin (100 nM) and N-tert-butyl-N cent -[(2 cyclohexylamino-5-nitrobenzene) sulfonyl] urea (BM-531, 10 microM) inhibited the spontaneous contractile activities of both longitudinal and circular muscles. The present results indicate that contractile TP receptors are present in the non pregnant porcine uterus. Therefore, the prostanoid receptor subtypes that exist in the porcine uterus (TP, IP, DP, FP, EP(1), EP(2) and EP(3)) are the same as those present in the human uterus. The distribution of TP receptors in the porcine uterus differed depending on the type of myometrium (longitudinal and circular muscles) and region of the uterus. The endogenous thromboxane A(2)-TP receptor pathway is thought to play a physiological role in regulation of spontaneous contractile activity in the porcine uterus. PMID- 14757157 TI - Antitussive activity of the tachykinin NK1 receptor antagonist, CP-99994, in dogs. AB - CP-99994 [(+)-(2S,3S)-3-(2-methoxybenzylamino)-2-phenylpiperidine] is a selective tachykinin NK(1) receptor antagonist that inhibits cough in guinea pigs and cats. This study examined the antitussive effects of CP-99994 in dogs produced by mechanical stimulation of the intrathoracic trachea. CP-99994 (10 mg/kg, p.o.) inhibited cough frequency by 52% at 2 h, 31% at 6 h and by 21% at 24 h. Cough amplitude was inhibited by 45% at 6 h but unchanged at 2 and 24 h after CP-99994. Plasma levels of CP-99994 were highest at 2 h (75+/-26 ng/ml) and fell to 22+/-6 ng/ml at 6 h. These results demonstrate antitussive activity of CP-99994 in dogs at a dose proven to antagonize tachykinin NK(1) receptors in this species. PMID- 14757158 TI - Discovery of a novel protein tyrosine phosphatase-1B inhibitor, KR61639: potential development as an antihyperglycemic agent. AB - Protein tyrosine phosphatase-1B (PTP-1B), a negative regulator of insulin signaling, may be an attractive therapeutic target for type 2 diabetes mellitus. High throughput screening (HTS) for PTP-1B inhibitors using compounds from the Korea Chemical Bank identified several hits (active compounds). Among them, a hit with 1,2-naphthoquinone scaffold was chosen for lead development. KR61639, [4-[1 (1H-indol-3-yl)-3,4-dioxo-3,4-dihydro-naphthalen-2-ylmethyl]-phenoxy]-acetic acid tert-butyl ester, inhibited human recombinant PTP-1B with an IC(50) value of 0.65 microM in a noncompetitive manner. KR61639 showed modest selectivity over several phosphatases and increased insulin-stimulated glycogen synthesis in HepG2 cells and stimulated 2-deoxyglucose uptake in 3T3/L1 adipocytes. In addition, in vivo study using ob/ob mouse demonstrated that KR61639 exerted a hypoglycemic action when given orally. Thus, KR61639 may be a good starting point for lead optimization in developing a novel antidiabetic agent. PMID- 14757159 TI - Mutation S363A in the human delta-opioid receptor selectively reduces down regulation by a peptide agonist. AB - Chemically distinct opioid agonists have different abilities to down-regulate opioid receptors. The present study investigated the role of Ser(363) in human delta-opioid receptor down-regulation by a delta-selective peptide- and non peptide agonist. Cyclic[D-Pen(2),D-Pen(5)]enkephalin (DPDPE)-mediated down regulation was significantly attenuated by a S363A mutation. In contrast, this mutation had no effect on down-regulation by (+)-4-[(alpha R)-alpha-((2S,5R)-4 allyl-2,5-dimethyl-1-piperazinyl)-3-methoxybenzyl]N,N-diethylbenzamide (SNC80). These results demonstrate that the molecular mechanism of the human delta-opioid receptor down-regulation is agonist-specific. PMID- 14757160 TI - Site-directed mutagenesis at the human B2 receptor and molecular modelling to define the pharmacophore of non-peptide bradykinin receptor antagonists. AB - Combining site-directed mutagenesis with information obtained from molecular modelling of the bradykinin (BK) human B2 receptor (hB2R) as derived from the bovine rhodopsin crystal structure [Science 289 (2000) 739], we previously defined a putative binding mode for the non-peptide B2 receptor antagonists, FR173657 and LF16-0687 [Can J Physiol Pharmacol 80 (2002) 303]. The present work is aimed to define the specific role of the quinoline moiety in the pharmacophore of these non-peptide antagonists. The effect of the mutations I110A, L114A (TM, transmembrane 3), W256A (TM6), F292A, Y295A and Y295F (TM7) was evaluated. None of the mutations affected the binding interaction of peptide ligands: the agonist BK and the peptide antagonist MEN 11270. The affinities in competing for [3H]-BK binding and in blocking the BK-induced IP production by the non-peptide antagonists LF16-0687 and FR173657 at the wild type and mutant receptors were analysed. While the affinities of LF16-0687 and FR173657 were crucially decreased at the I110A, Y295A, and Y295F mutants, the W256A mutation affected the affinity of the LF16-0687 only. The important contribution of the quinoline moiety was shown by the inability of an analogue of LF16-0687, lacking this moiety, to affect BK binding at the wild type receptor. On the other hand, the benzamidine group did not interact with mutated residues, since LF16-0687 analogues without this group or with an oxidated benzamidine displayed pairwise loss of affinity on wild type and mutated receptors. Further differences between FR173657 and LF16 0687 were highlighted at the I110 and Y295 mutants when comparing binding (pK(i)) and functional antagonist (pKB) affinity. First, the I110A mutation similarly impaired their binding affinity (250-fold), but at a less extent the antagonist potency of FR173657 only. Second, both the hydroxyl and the phenyl moieties of the Y295 residue had a specific role in the LF16-0687 interaction with the receptor, as demonstrated at the Y295F and Y295A mutants, respectively, but not in that of FR173657. Present data identify a receptor binding pocket comprised among TM3, 6, and 7, which concerns the interaction of the non-peptide antagonists FR173657 and LF16-0687, but not that of the peptide agonist or antagonist. Results indicate the quinoline group as the involved pharmacophoric element, and that the studied residues are differently involved in the interaction. The analysis performed by means of the GRID software led us to propose different spatial orientations of the quinoline moieties and partially overlapping binding pockets for the two ligands: that of LF16-0687 is located in the lipophilic environment amongst I110 (TM3), W256 (TM6), and Y295 (TM7) residues, whereas that of FR173657 lies essentially between I110 and Y295. PMID- 14757161 TI - Mitochondria permeability transition-dependent tert-butyl hydroperoxide-induced apoptosis in hepatoma HepG2 cells. AB - Tert-butyl hydroperoxide (t-BHP) has been demonstrated to induce apoptosis in hepatoma cell line HepG2, but poor data were available on the signaling pathway initiated by t-BHP. In this work, we studied in details the apoptotic pathways induced in HepG2 cells by t-BHP. DNA fragmentation, activation of caspases and cytochrome c release were demonstrated. Permeability transition pore inhibitors prevented the DNA fragmentation and caspase activation induced by t-BHP. In addition, changes in the mitochondrial membrane potential were detected: hyperpolarization preceded loss of membrane potential. It also preceded caspase activation which occurred before the induction of DNA fragmentation. Taken together, these results emphasize the central role played by mitochondria in the initiation of apoptosis in HepG2 cells exposed to oxidant agents. PMID- 14757162 TI - 2-ClATP exerts anti-tumoural actions not mediated by P2 receptors in neuronal and glial cell lines. AB - We investigated the effects of the ATP analogue and P2 receptor agonist 2-ClATP on growth and survival of different neuronal (PC12, PC12nnr5 and SH-SY5Y) and glial (U87 and U373) cell lines, by the use of direct count of intact nuclei, fluorescence microscopy, fluorescence-activated cell sorter analysis (FACS) and high pressure liquid chromatography (HPLC). 2-ClATP lowered the number of cultured PC12nnr5, SH-SY5Y, U87 and U373 cells to almost 5%, and of PC12 cells to about 35% after 3-4 days of treatment. EC(50) was in the 5-25 microM range, with 2-ClATP behaving as a cytotoxic or cytostatic agent. Analysis of the biological mechanisms demonstrated that pyridoxalphosphate-6-azophenyl-2',4'-disulphonic acid (P2 receptor antagonist and nucleotidases inhibitor), but not Caffeine or CGS-15493 (P1 receptor antagonists) effectively prevented 2-ClATP-induced toxicity. 2-ClATP metabolic products (2-ClADP, 2-ClAMP, 2-Cladenosine) and new synthesis derivatives (2-CldAMP, 2-Cldadenosine-3',5'-bisphosphate and 2-CldATP) exerted similar cytotoxic actions. Inhibition of both serum nucleotidases and purine nucleoside transporters strongly reduced 2-ClATP-induced cell death, which was conversely increased by the nucleotide hydrolyzing enzyme apyrase. The adenosine kinase inhibitor 5-iodotubericidin totally prevented 2-ClATP or 2 Cladenosine-induced toxicity. In summary, our findings indicate that 2-ClATP exerts either cell cycle arrest or cell death, acting neither on P2 nor on P1 receptors, but being extracellularly metabolized into 2-Cladenosine, intracellularly transported and re-phosphorylated. PMID- 14757163 TI - Antitumor AZA-anthrapyrazoles: biophysical and biochemical studies on 8- and 9 aza regioisomers. AB - Aza-bioisosteres of anthrapyrazoles (Aza-APs) bearing the C-N substitution at position 9 are powerful anticancer agents now in clinical trials. In contrast, their 8-substituted regioisomers are practically devoid of chemotherapeutic effects. To understand the molecular basis for a dramatically different response by otherwise very similar compounds, we performed a detailed investigation on the physico-chemical properties of several aza-APs belonging to the two families, on their DNA-binding affinity and specificity as well as on their capacity to impair the activity of the two isoforms of human Topoisomerase II (top2alpha and top2beta). Our results indicate that molecular size and shape, electronic distribution, redox properties, lipophilicity and protonation equilibria are essentially the same when comparing 9- with 8-substituted congeners. Although no major difference could be picked up when comparing the DNA binding properties of corresponding members of the 8- and 9-aza families, interestingly the affinity and specificity for the nucleic acid is modulated by the nature of the side-arms linked to the aza-AP scaffold, suggesting structural motifs that may determine DNA sequence recognition by the studied drug. Topoisomerase II poisoning activity was much higher for 9-aza derivatives than 8-aza analogues as shown by a cleavage assay with purified recombinant top2 isoforms. The difference appears to account for the divergent anticancer potential exhibited by different aza-AP regioisomers and suggests a specific molecular recognition of the cleavage complex by the studied drugs. PMID- 14757164 TI - Inhibition of plasmin-mediated prostromelysin-1 activation by interaction of long chain unsaturated fatty acids with kringle 5. AB - C18 unsaturated fatty acids were here found to inhibit proMMP (matrix metalloproteinase)-3 activation by plasmin. This effect was suppressed by lysine ligand competitors, indicating that it was mediated by binding to kringle domains. Surface plasmon resonance analysis demonstrated that oleic acid interacted to a similar extent with plasmin and kringle 5 (KD values of 3.4 x 10( 8) and 5.9 x 10(-8)M) while interaction with kringles 1-2-3 was 10-fold lower. Furthermore, oleic acid stimulated the amidolytic activity of plasmin and mini plasmin, but not micro-plasmin. Oleic acid also enhanced u-PA (urokinase-type plasminogen activator)-mediated plasminogen activation over 50-fold. Taken together, these data indicate that inhibition of plasmin-induced proMMP-3 activation by unsaturated fatty acids was mediated through their preferential binding to kringle 5. The influence of elaidic acid on the plasmin/MMP-3/MMP-1 proteolytic cascade was assessed ex vivo. Exogenous addition of plasmin to dermal fibroblasts or supplementation of gingival fibroblast culture medium with plasminogen triggered this cascade. In both instances, elaidic acid totally abolished proMMP-3 and proMMP-1 activation. Additionally, a significant decrease in lattice retraction and collagen degradation in a range similar to that obtained with Batimastat was observed when human gingival fibroblasts were cultured in plasminogen-containing type I collagen gels, indicative of the dual influence of unsaturated fatty acids on MMP activation and activity. In conclusion, unsaturated fatty acids or molecules with similar structures could be attractive target for the development of natural pharmacological inhibitors directed against plasmin and/or MMPs in different pathological contexts such, skin UV irradiation, vascular diseases and tumour growth and invasion. PMID- 14757165 TI - Isolation and identification of heterogeneous nuclear ribonucleoproteins (hnRNP) from purified plasma membranes of human tumour cell lines as albumin-binding proteins. AB - Since albumin is being developed as a drug carrier to target tumours the search for albumin-binding proteins (ABPs), which play a role in cell surface binding and endocytosis of native and conjugated albumins becomes more and more interesting. We isolated five different proteins from purified plasma membranes from three different human tumour cell lines (CCRF-CEM, MV3 and MCF7) by albumin affinity chromatography and identified them as four members of the heterogeneous nuclear ribonucleoproteins (hnRNP) family and calreticulin by matrix-assisted laser desorption ionisation time-of-flight mass spectrometry. Contamination of the plasma membrane preparation by nuclear membranes was excluded with anti nucleopore antibodies. Western blot analyses of plasma membranes showed ABPs with the same molecular weights as the albumin-affinity isolates. Tryptic digestion of intact cells was used to determine the sidedness of the albumin-binding property, which is oriented to the exterior of the cell. Localisation to the plasma membrane and albumin binding is a novel property of hnRNP. PMID- 14757166 TI - Molecular pharmacology of the ovine melatonin receptor: comparison with recombinant human MT1 and MT2 receptors. AB - The variations of the pharmacological properties of melatonin receptors between different mammalian species in transfected cell lines have been poorly investigated. In the present study, melatonin analogues have been used to characterize the pharmacology of the recombinant ovine melatonin receptor (oMT1) expressed in CHO cell lines and the native oMT1 from the pars tuberalis (PT). Studies with selective ligands on native and transfected oMT1 showed similar properties for binding affinities [r2(PT/CHO) = 0.85]. The affinities and the functional activities of these ligands were compared with the human receptors (hMT1 or hMT2) expressed in CHO cells as well. The oMT1 and hMT1 receptors had similar pharmacological profiles (r2=0.82). Nevertheless, some of the selective compounds at the human receptor presented a reduced affinity at the ovine receptor. Furthermore, some compounds showed marked different functional activities at oMT1 vs. hMT1 receptors. Our findings demonstrated differences in the pharmacological properties of melatonin receptors in ovine and human species. PMID- 14757167 TI - Specific ligand binding on genetic variants of human alpha1-acid glycoprotein studied by circular dichroism spectroscopy. AB - Human alpha1-acid glycoprotein displays genetic polymorphism. Different drug binding properties of the two main genetic products (F1-S and A variants) have been demonstrated. In search for specific circular dichroism (CD) probes, dicumarol and acridine orange were found to specifically bind to the F1-S and A variants, respectively. Dicumarol binding to the F1-S variant produced induced Cotton effects originating from the favored chiral conformation of the bound label. Acridine orange gave induced biphasic Cotton effects due to chiral intermolecular exciton interaction between label molecules bound to the A variant. Displacement of the CD probes by specific marker ligands was demonstrated. The induced CD spectrum of dicumarol was found to change sign in the presence of imipramine, as a manifestation of high-affinity ternary complex formation on the F1-S variant. PMID- 14757168 TI - Mitochondrial thioredoxin reductase inhibition by gold(I) compounds and concurrent stimulation of permeability transition and release of cytochrome c. AB - The effects of auranofin, chloro(triethylphosphine)gold(I) (TEPAu), and aurothiomalate on mitochondrial respiration, pyridine nucleotide redox state, membrane permeability properties, and redox enzymes activities were compared. The three gold(I) derivatives, in the submicromolar range, were extremely potent inhibitors of thioredoxin reductase and stimulators of the mitochondrial membrane permeability transition (MPT). Auranofin appeared as the most effective one. In the micromolar range, it inhibited respiratory chain and glutathione peroxidase activity only slightly if not at all. TEPAu and aurothiomalate exhibited effects similar to auranofin, although TEPAu showed a moderate inhibition on respiration. Aurothiomalate inhibited glutathione peroxidase at concentrations where auranofin and TEPAu were without effect. Under nonswelling conditions, the presence of auranofin and aurothiomalate did not alter the redox properties of the mitochondrial pyridine nucleotides indicating that membrane permeability transition occurred independently of the preliminary oxidation of pyridine nucleotides. Under the same experimental conditions, TEPAu showed a moderate stimulation of pyridine nucleotides oxidation. Mitochondrial total thiol groups, in the presence of the gold(I) derivatives, slightly decreased, indicating the occurrence of an oxidative trend. Concomitantly with MPT, gold(I) compounds determined the release of cytochrome c that, however, occurred also in the presence of cyclosporin A and, partially, of EGTA, indicating its independence of MPT. It is concluded that the specific inhibition of thioredoxin reductase by gold(I) compounds may be the determinant of MPT and the release of cytochrome c. PMID- 14757169 TI - Inhibition of glycogen breakdown by imino sugars in vitro and in vivo. AB - The imino sugar N-butyldeoxynojirimycin (NB-DNJ) is a glucose analogue which inhibits the glycoprotein N-glycan processing enzymes alpha-glucosidases I and II and the ceramide glucosyltransferase that catalyses the first step of glycosphingolipid biosynthesis. This and other N-alkylated DNJ compounds have the potential to inhibit other glucosidase, including acid alpha-glucosidase and alpha-1,6-glucosidase, enzymes involved in glycogen breakdown. We have investigated the effect of NB-DNJ and N-nonyldeoxynojirimycin (NN-DNJ) on glycogen catabolism. Both NB-DNJ and NN-DNJ were potent inhibitors of acid alpha glucosidase and alpha-1,6-glucosidase in vitro. NB-DNJ and NN-DNJ inhibited liver glycogen breakdown in vivo in fasting mice. Inhibition of glycogen catabolism occurred in the cytosol and lysosomes. The liver glycogen breakdown inhibition was only induced at high doses of NB-DNJ, whereas NN-DNJ caused glycogen accumulation at lower doses. The in vivo effect of NB-DNJ on liver glycogen was transient as there was no inhibition of breakdown after 90 days of treatment. The inhibition by NN-DNJ, was more pronounced, reached a plateau at 50 days and then remained unchanged. Increased glycogen was also observed in skeletal muscle in NB DNJ- and NN-DNJ-treated mice. Since the effects on glycogen metabolism by NB-DNJ are transient and only occur at high concentrations, it is not predicted that glycogen breakdown will be impaired in patients receiving NB-DNJ therapy. NN-DNJ is the prototype of long alkyl chain derivatives of DNJ that are entering pre clinical development as potential hepatitis B/hepatitis C (HBV/HCV) therapeutics. Depending on the dose of these compounds used, there is the potential for glycogen catabolism to be partially impaired in experimental animals and man. PMID- 14757170 TI - Benzylamide derivative compound attenuates the ultraviolet B-induced hyperpigmentation in the brownish guinea pig skin. AB - This study evaluated the effects of synthetic benzylamide compound I (2,6 dimethoxy-N-phenylbenzamide) on the ultraviolet B (UV B)-induced hyperpigmentation of the skin. UV B-induced hyperpigmentation was elicited on brownish guinea pig skin according to the method reported by Hideya et al. [Arch Dermatol Res 290 (1998) 375] with minor modifications. A lightening effect was observed following the topical application of compound I on UV-stimulated hyperpigmentation. The skin returned to its original color after treatment with compound I. Fontana-Masson staining indicated that melanin level in the hyperpigmented area was significantly decreased in the compound I-treated animals. However, the number of melanocytes were not changed in the compound I treated groups using the S-100 stain, which is an immunohistochemical method. In vitro experiments using the cultured melanoma cells showed a 31.7% inhibition of melanin production by compound I at 100 microM. In addition, this compound had no effect on the tyrosinase enzyme function. However, it exhibited a catalyzing effect on the dopachrome transformation into 5,6-dihydroxyindole-2-carboxylic acid. Overall, the pigment-lightening effects of the compound I may due to the dopachrome tautomerase stimulation. PMID- 14757171 TI - Genistein arrests hepatoma cells at G2/M phase: involvement of ATM activation and upregulation of p21waf1/cip1 and Wee1. AB - Genistein, a soy isoflavone, has a wide range of biological actions that suggest it may be of use in cancer prevention. We have recently reported that it arrests hepatoma cells at G2/M phase and inhibits Cdc2 kinase activity. In the present study, we examined the signaling pathway by which genistein modulates Cdc2 kinase activity in HepG2 cells and leads to G2/M arrest, and found that it caused an increase in both Cdc2 phosphorylation and expression of the Cdc2-active kinase, Wee1. Genistein also enhanced the expression of the cell cycle inhibitor, p21waf1/cip1, which interacts with Cdc2. Furthermore, phosphorylation/inactivation of Cdc25C phosphatase, which dephosphorylates/activates Cdc2, was increased. Genistein enhanced the activity of the checkpoint kinase, Chk2, which phosphorylates/inactivates Cdc25C, induced accumulation of p53, and activated the ataxia-telangiectasia-mutated (ATM) gene. Caffeine, an ATM kinase inhibitor, inhibited these effects of genistein on Chk2, p53, and p21waf1/cip1. These findings suggest that the effect of genistein on G2/M arrest in HepG2 cells is partly due to ATM-dependent Chk2 activation, an increase in Cdc2 phosphorylation/inactivation as a result of induction of Wee1 expression, and a decrease in Cdc2 activity as a result of induction of p21waf1/cip1 expression. PMID- 14757172 TI - Investigation of ouabain-induced anticancer effect in human androgen-independent prostate cancer PC-3 cells. AB - To determine the therapeutic potential of cardiac glycosides in androgen independent prostate cancer, we examined ouabain-induced cytotoxic effect as well as the signaling pathways in PC-3 cells. Ouabain induced a time- and concentration-dependent cytotoxicity using mitochondrial MTT reduction assays, and the effective threshold concentration was in nanomolar level. At the concentrations less than 10 nM, ouabain induced a decrease of mitochondrial activity until a 7-hr exposure was performed, while it induced a rapid drop of mitochondrial function as early as a 2-hr treatment of cells with high concentrations of ouabain suggesting the involvement of two distinct mechanisms to ouabain action. After functional examinations, the data showed that both low and high concentrations of ouabain induced an inhibition of Na+-K+ ATPase and a subsequent 45Ca2+ influx into PC-3 cells. High concentrations of ouabain induced a significant and time-dependent loss of mitochondrial membrane potential (Deltapsim), a sustained production of reactive oxygen species (ROS), and severe apoptotic reaction. Ouabain also induced an increase of Par-4 (prostate apoptosis response 4) expression. Furthermore, an antisense, but not nonsense, oligomer against Par-4 expression significantly inhibited the cytotoxicity induced by low concentrations of ouabain. It is suggested that ouabain induces two modes of cytotoxic effect in human hormone-independent prostate cancer PC-3 cells. Low concentrations of ouabain induce the increase of Par-4 expression and sensitize the cytotoxicity; while high concentrations of ouabain induce a loss of Deltapsim, a sustained ROS production and a severe apoptosis in PC-3 cells. PMID- 14757173 TI - The development of morphine antinociceptive tolerance in nitric oxide synthase deficient mice. AB - Elevations in nitric oxide (NO) have been implicated in the development of morphine antinociceptive tolerance. This study was conducted to establish the role of specific isoforms of NO synthase (NOS) in morphine tolerance development using genetically modified mice. METHODS: Three groups of mice (endothelial NOS [eNOS]-deficient, neuronal NOS [nNOS]-deficient, and NOS-competent) were used in this experiment. On Day 1, the analgesic response (radiant heat tail-flick) to a challenge dose of morphine (4 mg/kg) was determined over 3 hr. Tolerance was induced on Days 1-5 by administering morphine subcutaneously (10 mg/kg) or L arginine, a NO precursor, intraperitoneally (200 mg/kg), twice daily. Analgesic response to the challenge dose was determined again on Day 6. RESULTS: Following sustained morphine administration, nNOS-deficient mice exhibited less tolerance development when compared to the control group, although measurable tolerance still occurred. Mice deficient in eNOS evidenced a degree of tolerance similar to that of control. Prolonged L-arginine administration produced significant functional tolerance to morphine in NOS-competent and eNOS-deficient mice. The loss of morphine responsivity after L-arginine administration was similar to that after morphine pretreatment. L-Arginine did not affect the antinociceptive response to morphine in mice deficient in nNOS, suggesting that the small degree of morphine-induced tolerance in this group occurs through an alternate pathway. CONCLUSIONS: These data demonstrate the pivotal role of the neuronal isoform of NOS in development of morphine antinociceptive tolerance. Furthermore, tolerance development appears to be predominantly a NO-mediated process, but likely is augmented by a secondary (non-NO) pathway. PMID- 14757174 TI - Relationship between the antinociceptive response to desipramine and changes in GABAB receptor function and subunit expression in the dorsal horn of the rat spinal cord. AB - Although tricyclic antidepressants are among the drugs of choice for the treatment of neuropathic pain, their mechanism of action in this regard remains unknown. Because previous reports suggest these agents may influence gamma aminobutyric acid (GABA) neurotransmission, and GABAB receptors are known to participate in the transmission of pain impulses, the present experiments were undertaken to examine whether the administration of desipramine alters GABAB receptor subunit expression and function in the dorsal horn of the rat spinal cord. For the study, rats were injected (i.p.) once daily with desipramine (15 mg/kg) for 7 consecutive days, during which their thermal withdrawal threshold was monitored, and after which GABAB receptor function, and the levels of GABAB receptor subunit mRNA, were quantified in the spinal cord dorsal horn. The results indicate that 4-7 days of continuous administration of desipramine are necessary to observe a significant increase in the thermal pain threshold. Moreover, it was found that 7 days of treatment with desipramine enhances GABAB receptor function, as measured by baclofen-stimulated [35S]GTPgammaS binding, and increases mRNA expression for the GABAB(1a) and GABAB(2), but not GABAB(1b), subunits. These findings suggest the antinociceptive effect of desipramine is accompanied by a change in spinal cord GABAB receptor sensitivity that could be an important component in the analgesic response to this agent. PMID- 14757175 TI - Nicotine-related alkaloids and metabolites as inhibitors of human cytochrome P 450 2A6. AB - S-(-)-Nicotine and 13 of the most prevalent nicotine-related alkaloids and metabolites (i.e., S-(-)-nornicotine, myosmine, beta-nicotyrine, S-cotinine, S norcotinine, S-(-)-nicotine N-1'-oxide, S-(-)-nicotine Delta1'-5'-iminium ion, S (-)-anabasine, S-(-)-N-methylanabasine, anabaseine, S-(-)-anatabine, nicotelline, and 2,3'-bipyridyl) were evaluated as inhibitors of human cDNA-expressed cytochrome P-450 2A6 (CYP2A6) mediated coumarin 7-hydroxylation. Tobacco alkaloids myosmine, S-(-)-nornicotine, S-cotinine, S-norcotinine, S-(-)-nicotine N-1'-oxide, S-(-)-nicotine Delta1'-5'-iminium ion, S-(-)-N-methylanabasine, anabaseine, and nicotelline had Ki values for inhibition of coumarin 7 hydroxylation ranging from 20 microM to more than 300 microM whereas nicotine and S-(-)-anatabine were much more potent (i.e. 4.4 and 3.8 microM, respectively). The tobacco alkaloids 2,3'-bipyridyl (7.7 microM) and S-(-)-anabasine (5.4 microM), were somewhat less potent compared with S-(-)-nicotine or S-(-) anatabine in inhibition of human CYP2A6. beta-Nicotyrine, in which the N methylpyrrolidino moiety of nicotine was replaced by the aromatic N-methylpyrrole ring, was shown to inhibit human CYP2A6 with much greater potency (Ki=0.37 microM) compared with S-(-)-nicotine. Among the compounds examined, only nicotine and beta-nicotyrine were mechanism-based inhibitors of human CYP2A6. The potency of the mechanism-based CYP2A6 inhibitors suggests that, for smokers, modulation of CYP2A6 may be greater than that predicted on the basis of serum concentration of these alkaloids. Our results indicate that the prominent nicotine-related alkaloid beta-nicotyrine present after smoking potently inhibits human CYP2A6. PMID- 14757176 TI - Gene expression profile of the Gs-coupled prostacyclin receptor in human vascular smooth muscle cells. AB - Migration and proliferation of medial smooth muscle cells (SMC) in the arterial intima contributes to the development of atherosclerotic plaques and restenotic processes after coronary angioplasty. Prostacyclin (PGI2)-mediated stimulation of cyclic adenosine 3'5'-monophosphate (cAMP) signaling is believed to be important for maintaining SMC in a quiescent state. In order to identify new cellular targets of PGI2/cAMP action, we have used microarray screening to examine changes in the transcriptional profile in human vascular SMC in response to exposure to the stable PGI2 mimetic iloprost. We have identified 83 genes with significantly altered expression after iloprost (100 nM) exposure for 6 hr. Fifty-one genes were upregulated, among them stanniocalcin precursor (18.8+/-2.7), zinc finger transcription factor (7.8+/-2.0), hyaluronan synthase 2 (6.8+/-1.8), cyclooxygenase 2 (4.7+/-0.8), dual specific phosphatase (3.9+/-0.5) and vascular endothelial growth factor (2.3+/-0.4). Thirty-two genes were reduced, among them cystein-rich angiogenic protein (-14.9+/-1.3), monocyte chemotactic protein 1 ( 7.4+/-1.1) and plasminogen activator inhibitor PAI-1 (-4.5+/-0.5). By means of semi-quantitative RT-PCR, time-courses of gene expression were established. The present study identified genes not hitherto recognized to be targets of PGI2 action, providing further insight into its cAMP-mediated effects on SMC growth, migration and matrix secretion. PMID- 14757177 TI - Regulation of inosine monophosphate dehydrogenase type I and type II isoforms in human lymphocytes. AB - The enzyme inosine monophosphate dehydrogenase (IMPDH) catalyzes the rate limiting step in the de novo biosynthesis of guanine nucleotides. Inhibition of IMPDH leads to immunosuppression by decreasing guanine nucleotides that are required for the proliferation of lymphocytes. IMPDH activity is mediated by two highly conserved isoforms, type I and type II. We have characterized the mRNA and protein expression of the two isoforms in a variety of human tissues, peripheral blood mononuclear cells (PBMCs), and selected cell lines to investigate their regulation. Type I mRNA was expressed in most tissues with high expression in PBMCs and low expression in thymus. IMPDH type II transcript was also detected in most tissues with low expression in spleen and PBMCs. In PBMCs, induction of both type I and type II mRNAs was observed within 12 hr of mitogenic stimulation. Using type-selective IMPDH antibodies, an increase in the levels of type I and type II proteins was observed after mitogenic stimulation. The effect of two IMPDH inhibitors, MPA and VX-497, was investigated on the expression of type I and type II isoforms. VX-497 is an orally bioavailable, potent and reversible inhibitor of IMPDH, with broad applicability in many viral and immune system mediated diseases. MPA and VX-497 inhibit both isoforms of IMPDH in vitro. Prolonged treatment of lymphocytes with either VX-497 or MPA did not lead to an increase in type I or type II IMPDH protein levels. These results are discussed in the context of IMPDH being a target for immunosuppressive, anti-viral and anti cancer therapy. PMID- 14757178 TI - Isorhapontigenin and resveratrol suppress oxLDL-induced proliferation and activation of ERK1/2 mitogen-activated protein kinases of bovine aortic smooth muscle cells. AB - The objective of our study was to compare the inhibitory effect of isorhapontigenin (ISO) and resveratrol, two natural antioxidants, on oxidized low density lipoprotein (oxLDL)-induced proliferation of bovine aortic smooth muscle cells (BASMCs) and its relation to reactive oxygen species (ROS) generation and extracellular signal-regulated kinase 1/2 activation. The results showed that stimulation of oxLDL (50-150 microg/mL) for 48 hr induced a dose-dependent increase in cell number and incorporation of [3H]thymidine into DNA of BASMCs. Western blot analysis demonstrated that oxLDL (150 microg/mL) stimulated an evident phosphorylation of p42/44 MAP kinases in BASMCs. Incubation of BASMCs with oxLDL induced significant increase in ROS detected by using an oxidant sensitive fluorescent probe of 2',7'-dichlorofluorescin diacetate. The level of H2O2 in the medium of cultured BASMCs also increased markedly. Preincubation of BASMCs with ISO and resveratrol significantly inhibited oxLDL-induced cell proliferation and incorporation of [3H]thymidine, and the phosphorylation of p42/44 MAP kinases in BASMCs as well. Furthermore, preincubation of BASMCs with ISO and resveratrol attenuated oxLDL-induced increases in ROS and H2O2 levels. The results suggested that oxLDL-induced acute formation of ROS and subsequent activation of redox-sensitive extracellular signal-regulated kinase 1/2 MAPK pathways, which might be important for mitogenic signaling of oxLDL in vascular smooth muscle cells. The inhibitory effect of ISO and resveratrol on oxLDL induced mitogenesis of BASMCs might be taken through blocking the generation of ROS and activation of the ERKs pathway. PMID- 14757179 TI - Inhibition of nitric oxide/cyclic GMP-mediated relaxation by purified flavonoids, baicalin and baicalein, in rat aortic rings. AB - The dried roots of Scutellaria baicalensis Georgi (Huangqin) are widely used in traditional Chinese medicine. We purified two flavonoids, baicalin and baicalein from S. baicalensis Georgi and examined their effects on isolated rat aortic rings. Baicalin (3-50 microM) inhibited endothelium/nitric oxide (NO)-dependent relaxation induced by acetylcholine (Ach) or cyclopiazonic acid (CPA). Baicalein at 50 microM abolished Ach-induced relaxation and markedly reduced CPA-induced relaxation. Treatment with 1mM L-arginine partially but significantly reversed the effects of baicalin (50 microM) or baicalein (50 microM) on Ach-induced relaxation. In endothelium-denuded rings, treatment with baicalin, baicalein or methylene blue partially inhibited relaxations induced by the NO donors, sodium nitroprusside (SNP) and hydroxylamine. Both flavonoids markedly reduced the increase in cyclic GMP levels stimulated by Ach in endothelium-intact rings and by SNP in endothelium-denuded rings. In contrast, exposure of endothelium-denuded rings to baicalin or baicalein did not affect relaxations induced by pinacidil or NS 1619, putative K+ channel activators. Neither flavonoids affected agonist induced increase in the endothelial [Ca2+]i. Our results indicate that baicalin and baicalein attenuated NO-mediated aortic relaxation and cyclic GMP increases, likely through inhibition of NO-dependent guanylate cyclase activity. PMID- 14757180 TI - Cross talk of cAMP and flavone in regulation of cystic fibrosis transmembrane conductance regulator (CFTR) Cl- channel and Na+/K+/2Cl- cotransporter in renal epithelial A6 cells. AB - We have reported that in renal epithelial A6 cells flavones stimulate the transepithelial Cl- secretion by activating the cystic fibrosis transmembrane conductance regulator (CFTR) Cl- channel and/or the Na+/K+/2Cl- cotransporter. On the other hand, it has been established that cAMP activates the CFTR Cl- channel and the Na+/K+/2Cl- cotransporter. However, no information is available on the interaction between cAMP and flavones on stimulation of the CFTR Cl- channel and the Na+/K+/2Cl- cotransporter. To clarify the interaction between cAMP and flavones, we studied the regulatory mechanism of the CFTR Cl- channel and the Na+/K+/2Cl- cotransporter by flavones (apigenin, luteolin, kaempherol, and quercetin) under the basal and cAMP-stimulated conditions in renal epithelial A6 cells. Under the basal (cAMP-unstimulated) condition, these flavones stimulated the Cl- secretion by activating the Na+/K+/2Cl- cotransporter without any significant effects on the CFTR Cl- channel activity. On the other hand, these flavones diminished the activity of the cAMP-stimulated Na+/K+/2Cl- cotransporter without any significant effects on the CFTR Cl- channel activity. Interestingly, the level of the flavone-induced Cl- secretion under the basal condition was identical to that under the cAMP-stimulated condition. Based on these results, it is suggested that although both cAMP and flavones activate the Na+/K+/2Cl- cotransporter, these flavones have more powerful effects than cAMP on the Na+/K+/2Cl- cotransporter. PMID- 14757182 TI - The effect of blood sampling method on indicators of physiological stress in reindeer (Rangifer tarandus tarandus). AB - The effects of manual blood sampling and remote blood sampling using automatic blood sampling equipment (ABSE) on plasma cortisol and catecholamine concentrations were studied on eight adult female reindeer (Rangifer tarandus tarandus). Contemporary body temperatures and heart rates were also recorded to determine their utility as other possible stress indicators. The animals were blood sampled once every hour with ABSE on 9-10 May and then by manual blood sampling on 13-14 May. Animals were also fitted with equipment to record heart rate and body temperature. Heart rate and body temperature were also recorded continuously without blood sampling on 17-18 May in undisturbed control conditions. Plasma cortisol concentrations were five-to-six fold greater during manual blood sampling compared to sampling with ABSE (F(1,3) = 13.34, P < 0.05). Plasma noradrenaline concentrations were significantly higher (F(1,3) = 22.98, P < 0.05) during manual blood sampling compared to sampling with ABSE, whereas plasma adrenaline concentrations did not differ. Heart rate was higher during manual blood sampling compared to control values. Body temperature was significantly higher during manual sampling compared to values recorded without blood sampling (F(1,4)= 31.65, P < 0.01). In conclusion, plasma cortisol concentration provides an excellent indicator of handling stress in reindeer. The use of ABSE for blood sampling enables measurements of plasma cortisol levels close to basal concentrations that may be used for reference values in studies where indicators of physiological stress are required. PMID- 14757183 TI - Parathyroid gland function in the uremic rabbit. AB - Rabbits with renal failure have been reported to be hypercalcemic and to have decreased parathyroid hormone (PTH) concentrations. Thus, it would seem that uremic rabbits are resistant to secondary hyperparathyroidism (HPT). The work reported here was designed to investigate parathyroid gland function in uremic rabbits and the effect of diets with different calcium (Ca) and phosphorus (P) content. The relationship between PTH and ionized calcium (Ca2+), parathyroid gland size and parathyroid cell cycle were studied in three groups of rabbits: Group I, rabbits with normal renal function on a standard diet (Ca = 1.2%, P = 0.6%); Group II, partially nephrectomized rabbits on a standard diet; and Group III, partially nephrectomized rabbits on a low Ca (0.6%)-high P (1.2%) diet. Group I rabbits had baseline Ca2+ = 1.71 +/- 0.05 mmol/l and PTH = 26.9 +/- 3.2 pg/ml. During hypo- and hypercalcemic stimulation PTH reached maximal values (PTHmax) of 94.4 +/- 5.5 pg/ml and minimal concentrations (PTHmin) of 3.2 +/- 0.2 pg/ml. Rabbits from Group II were hypercalcemic (baseline Ca2+ = 2.03 +/- 0.06 mmol/l) and had very low PTH levels (1.7 +/- 0.5 pg/ml); however, they reached a PTHmax that was similar to Group I, 92 +/- 8.7 pg/ml. Group III rabbits were hypocalcemic (baseline Ca2+ = 1.22 +/- 0.08 mmol/l) and had very high basal PTH levels (739 +/- 155 pg/ml). Their PTHmax and PTHmin were 801 +/- 169.4 pg/ml and 102.2 +/- 22.2 pg/ml, respectively. Both parathyroid gland size and parathyroid cell proliferation were increased in Group III. In conclusion, our results show that the Ca and P content of the diet markedly influence PTH secretion in the uremic rabbit and that when placed on a low Ca-high P diet uremic rabbits develop secondary HPT. PMID- 14757184 TI - Short-term changes of mRNA expression of various inflammatory factors and milk proteins in mammary tissue during LPS-induced mastitis. AB - During mammary gland infection, non-specific responses are the predominant ones. The goal of this study was to investigate the mRNA expression of various soluble immune components and of the major milk proteins during the acute phase of mammary inflammation. Five healthy lactating cows were intramammary infused in one quarter with 100 microg Escherichia coli-endotoxin (lipopolysaccharide, LPS) and the contralateral quarter with saline (9 g/l) serving as control. Mammary biopsy samples of both quarters were taken immediately before and at 3, 6, 9 and 12 h after infusion and mRNA expression of various factors was quantified via real-time RT-PCR. Blood samples for determination of leukocyte number were taken simultaneously with the biopsy samples and rectal temperature was measured at 1-h intervals. Rectal temperature increased until 5h (P < 0.05) after LPS administration and remained elevated until 9 h after LPS inoculation. Blood leukocyte number decreased (P < 0.05) from 0 to 3 h from 7.7 +/- 1.1 x 10(9)l(-1) to 5.7 +/- 1.0 x 10(9)l(-1) and thereafter recovered to pre-treatment levels until 12 h after LPS challenge. In LPS-treated quarters, tumor necrosis factor alpha and cyclooxygenase-2-mRNA expression increased (P < 0.05) to highest values at 3h after LPS challenge. Lactoferrin, lysozyme, inducible nitric oxide synthase increased (P < 0.05) and peaked at 6 h after challenge, and platelet-activating factor acetylhydrolase-mRNA expression tended to increase (P = 0.07). mRNA expression of insulin-like growth factor-I and of alphaS1-casein (CN), alphaS2 CN, beta-CN and beta-lactoglobulin did not change significantly, whereas mRNA expression of 5-lipoxygenase and alpha-lactalbumin decreased (P < 0.05) in both quarters and that of kappa-CN only in the LPS quarter. mRNA expression of some investigated factors (tumor necrosis factor-alpha, lysozyme, 5-lipoxygenase, alpha-lactalbumin) changed in control quarters, however in all respective factors less than in the LPS quarters (P < 0.05). In conclusion, mRNA expression of most inflammatory factors increased within hours, whereas that of most milk proteins remained unchanged. PMID- 14757185 TI - Early steps of insulin receptor signaling in chicken and rat: apparent refractoriness in chicken muscle. AB - The early steps of insulin receptor (IR) signaling (tyrosine phosphorylation of IR beta-subunit, IRS-1 and Shc and PI 3'-kinase activity) have been characterized in two target tissues in the chicken: liver and muscle. The signaling cascade appeared to depend on nutritional status in the liver, but not in muscle (with a possible exception for a minor tyrosine phosphorylation of the 52 kDa Shc isoform). In this study, we compared the responses of the liver and muscle to exogenous insulin (10 or 1000 mU/kg) in chickens and rats. In the liver, IRS-1 and Shc proteins were present in smaller amounts and the regulatory subunit p85 of PI 3'-kinase was present in larger amounts in chickens than in rats. In the basal state (saline injection), the level of tyrosine phosphorylation of IR was lower, and that of Shc higher, in chickens than in rats. PI 3'-kinase activity in chickens was half that in rats. Insulin activated all components of the cascade in a dose-dependent manner in both species. A different pattern was observed in the muscle. In the basal state, the levels of tyrosine phosphorylation of IR and of PI 3'-kinase activity were much higher in chickens than in rats (by factors of 2 and 30, respectively). Insulin strongly activated all components of the cascade in rats (but with no significant increase in the phosphorylation of Shc). No activation was observed in chickens (with only a slight but significant increase in the tyrosine phosphorylation of Shc). The insulin cascade therefore appears to respond normally in chicken liver but to be refractory in chicken muscle. The large amount of p85 and high levels of PI 3'-kinase activity in muscle may contribute to this situation, making chicken muscle an interesting model of insulin resistance. PMID- 14757186 TI - Nutrient supply enhances both IGF-I and MSTN mRNA levels in chicken skeletal muscle. AB - Nutrient supply may control muscle growth directly and indirectly through its influence on regulatory factors. The present study focuses on its effects on muscle insulin-like growth factors (IGF-I and -II) and myostatin (MSTN). Their mRNA levels were quantified by real time RT-PCR in pectoralis major (PM) and sartorius (SART) muscles from broiler chickens submitted to different feeding regimens (fed or fasted for 48 h) between hatch and 2 days of age and at 4 weeks of age. In the PM of 4 weeks old broilers, mRNA levels were also evaluated after a 16 h-fast and a refeeding period (refed 24 or 48 h after a 48 h-fast). In the PM muscle, both IGF-I and MSTN mRNA levels increased between 0 and 2 days of age in the fed group, while they remained low in the unfed one. A comparable trend was observed in the SART, but with lesser amplitude. In both muscles of 4 weeks old chickens, a 48 h-fast induced a significant reduction in MSTN mRNA levels (20% of fed state). In the PM, this effect required more than 16 h of fasting to occur and was fully reversed by only 24h of refeeding. IGF-I mRNA levels also varied with nutritional state. They decreased significantly with fasting in the SART muscle. By contrast, IGF-II mRNA levels did not vary significantly. Our data shows for the first time that two major paracrine regulators of muscle growth, IGF-I and MSTN, are sensitive to nutrient supply in hatching chicks, and also that fasting reduced IGF-I and MSTN mRNA levels in muscles of older chickens. PMID- 14757187 TI - Effects of colostrum feeding and dexamethasone treatment on mRNA levels of insulin-like growth factors (IGF)-I and -II, IGF binding proteins-2 and -3, and on receptors for growth hormone, IGF-I, IGF-II, and insulin in the gastrointestinal tract of neonatal calves. AB - The somatotropic axis regulates growth of the gastrointestinal tract (GIT). In addition, colostrum feeding and glucocorticoids affect maturation of the GIT around birth in mammals. We have measured mRNA levels of members of the somatotropic axis to test the hypothesis that colostrum intake and dexamethasone treatment affect respective gene expression in the GIT. Calves were fed either colostrum or an isoenergetic milk-based formula, and in each feeding group, half of the calves were treated with dexamethasone (DEXA; 30 microg/kg body weight per day). Individual parameters of the somatotropic axis differed (P < 0.05) among different GIT sections and formula feeding increased (P < 0.05) mRNA levels of individual parameters at various sites of the GIT. Effects of DEXA on the somatotropic axis in the GIT partly depended on different feeding. In colostrum fed calves, DEXA decreased (P < 0.05) mRNA levels of IGF-I (esophagus, fundus, duodenum, and ileum), IGF-II (fundus), IGFBP-2 (fundus), IGFBP-3 (fundus), IGF1R (esophagus, ileum, and colon), IGF2R (fundus), GHR (fundus), and InsR (esophagus, fundus), but in formula-fed calves DEXA increased mRNA levels of IGF-I (esophagus, rumen, jejunum, and colon). Furthermore, DEXA increased (P < 0.05) mRNA levels of IGF-II (pylorus), IGFBP-3 (duodenum), IGF2R (pylorus), and GHR (ileum), but decreased mRNA levels of IGFBP-2 (ileum), and IGF1R (fundus). Whereas formula feeding had stimulating effects, effects of DEXA treatment on the gene expression of parameters of the somatotropic axis varied among GIT sites and partly depended on feeding. PMID- 14757188 TI - Phosphorylation and nuclear accumulation are distinct events contributing to the activation of p53. AB - It has been recently shown that ionizing radiation (IR) and the mRNA synthesis inhibitor 5,6-dichloro-1-b-D-ribofuranosylbenzimidazole (DRB) act in synergy to induce p53-mediated transactivation of reporter plasmids in human cells [Oncogene 19 (2000) 3829]. We have extended these studies and show that ionizing radiation and DRB also act in synergy to induce ATM-mediated phosphorylation of the ser15 site of p53 and enhance the expression of endogenous p21 protein. Examination of the localization of p53 revealed that while DRB did not induce phosphorylation of the ser15 site of p53 but efficiently accumulated p53 in the nucleus, ionizing radiation induced phosphorylation of the ser15 site of p53 without prolonged nuclear accumulation. Importantly, the combination of DRB and IR resulted in a strong accumulation of phosphorylated p53 in the nucleus that was more persistent then p53 accumulation after IR alone. Furthermore, the nuclear export inhibitor leptomycin B showed a similar synergy with IR as did DRB regarding ser15 phosphorylation of p53 and p21 induction. These results suggest that the synergistic activation of the p53 response by the combination treatment is due to the activation of two distinct pathways where DRB causes the prolonged nuclear accumulation of p53 while ionizing radiation activates p53 by ATM-mediated phosphorylation. PMID- 14757189 TI - Developing azo and formazan dyes based on environmental considerations: Salmonella mutagenicity. AB - In previous papers, the synthesis and chemical properties of iron-complexed azo and formazan dyes were reported. It was shown that in certain cases iron could be substituted for the traditionally used metals such as chromium and cobalt, without having an adverse effect on dye stability. While these results suggested that the iron analogs were potential replacements for the commercially used chromium and cobalt prototypes, characterization of potentially adverse environmental effects of the new dyes was deemed an essential step in their further development. The present paper provides results from using the Salmonella/mammalian microsome assay to determine the mutagenicity of some important commercial metal complexed dyes, their unmetallized forms, and the corresponding iron-complexed analogs. The study compared the mutagenic properties of six unmetallized azo dyes, six commercial cobalt- or chromium-complexed azo dyes, six iron-complexed azo dyes, six unmetallized formazan dyes, and six iron complexed formazan dyes. The results of this study suggest that the mutagenicity of the unmetallized dye precursors plays a role in determining the mutagenicity of the iron-complexes. For the monoazo dye containing a nitro group, metal complex formation using iron or chromium decreased or removed mutagenicity in TA100; however, little reduction in mutagenicity was noted in TA98. For the formazan dye containing a nitro group, metal-complex formation using iron increased mutagenicity. Results varied for metal-complexes of azo and formazan dyes without nitro groups, but in general, the metal-complexed dyes based on mutagenic ligands were also mutagenic, while those dyes based on nonmutagenic ligands were nonmutagenic. PMID- 14757190 TI - Effects of oxidative and alkylating damage on microsatellite instability in nontumorigenic human cells. AB - Microsatellite instability is a phenomenon that is well characterized in mismatch repair-deficient tumor cell lines, including the potential etiological role of endogenous DNA damage. However, our understanding of microsatellite mutational mechanisms in repair-proficient, nontumorigenic cells is limited. We determined microsatellite mutation frequencies for human lymphoblastoid cells using an episomal DNA shuttle vector in which a (TTCC/AAGG)(9) microsatellite is inserted in-frame within the herpes simplex virus thymidine kinase (HSV-tk) gene. The responses of plasmid-bearing cells to reactive oxygen species or alkylating agents were compared after treatment with hydrogen peroxide (H(2)O(2)) and N ethyl-N-nitrosourea (ENU). H(2)O(2) treatment induced a statistically significant increase in overall HSV-tk mutation frequency relative to controls, with catalase reducing the effect. H(2)O(2) treatment increased the mutation frequency within the microsatellite and the HSV-tk coding region to a similar extent (five and six fold, respectively, relative to the control). Mutational specificity analyses demonstrated that the proportion of mutations within the microsatellite is not statistically different among the H(2)O(2), catalase, and PBS treatment groups. In contrast, treatment of cells bearing the microsatellite vector with ENU altered the mutational spectrum, relative to solvent control. ENU induced the expected base substitutions within the HSV-tk coding region, but did not increase the microsatellite mutation frequency. The low level of microsatellite mutagenesis observed after reactive oxygen species (ROS) insult likely reflects the normal repair processes of these nontumorigenic, repair-competent cells. Our ex vivo experiments demonstrate the manner in which repetitive DNA in normal human cells might respond to endogenous mutagens. PMID- 14757191 TI - Cell death evaluation in benzo[a]pyrene-transformed human breast epithelial cells after microcell-mediated transfer of chromosomes 11 and 17. AB - The incidence of apoptosis and nuclear instability, including the incidence of catastrophic death, were investigated in benzo[a]pyrene (BP)-transformed human breast epithelial cells (BP1-E cell line) after microcell-mediated transfer of chromosomes 11 and 17. Since the introduction of normal chromosomes 11 and 17 into tumorigenic human breast BP1-E cells reverts some of these cells' characteristics (especially those affected by microsatellite instabilities and loss of heterozygosity) to those of parental non-transformed MCF-10F cells, it was expected that the cell death rates would also be affected by this treatment. The transfer of the mentioned chromosomes, especially chromosome 17, to tumorigenic BP1-E cells increased the apoptotic ratios and decreased the nuclear instability ratios, thus showing that the microsatellite instability and loss of heterozygosity induced by BP in these chromosomes of MCF-10F cells affect the control of cell death mechanisms. PMID- 14757192 TI - Evidence of association of the CYP2E1 genetic polymorphism with micronuclei frequency in human peripheral blood. AB - Micronuclei (MN) are used as one of the cytogenetic biomarkers, and intra- and inter-individual variations in this frequency have been reported in human blood lymphocytes. Polymorphisms in a few metabolic enzyme genes seem to account for a proportion of this variability, but the impacts of specific genetic variants on the MN frequency have not yet been clarified. Here, we investigated the relationship between the MN frequency and several gene polymorphisms in 90 healthy Japanese men. The subjects with the CYP2E1(*)3 variant allele had a statistically lower mean MN frequency than subjects with the CYP2E1(*)1/(*)1 wild type. Furthermore, the adjusted odds ratio (OR) of the CYP2E1(*)3 variant with higher MN frequency levels was also significantly lower and calculated to be 0.25 (95% CI 0.07-0.83), when the OR for the subjects with the CYP2E1(*)1/(*)1 wild type was defined as 1.00. These data suggest that the CYP2E1(*)3 polymorphism may have the potential to influence the baseline frequency of MN. PMID- 14757193 TI - Aphidicolin and bleomycin induced chromosome damage as biomarker of mutagen sensitivity: a twin study. AB - The induction of chromosome damage in cultured human lymphocytes by in vitro treatments with aphidicolin (APC) and bleomycin (BLM) has been proposed as test of sensitivity to mutagens. To assess their validity, we have investigated whether the individual expression of induced chromosome damage has a genetic rather than an environmental basis. Metaphase analysis for chromosomal aberrations (CA) and micronucleus (MN) assay in cytokinesis-blocked cells have been performed in peripheral blood lymphocytes from 19 healthy male twins (9 monozygotic and 10 dizygotic pairs), aged 70-78 years, after APC, BLM and APC+BLM treatments. Concordance between twins revealed a high genetic component in the sensitivity towards clastogenic action of APC both as percentages of CA and MN. The micronucleus assay demonstrated a genetic basis also in the expression of chromosome damage induced by BLM and APC+BLM treatments. Since twins were elderly people, to investigate the possible role of age, CA and MN frequencies were compared with those found in lymphocytes from 11 young male donors. Basal and APC induced chromosome damage were clearly increased in the former. Following BLM and APC+BLM treatments, age significantly increased mitotic delay, as shown by the mitotic indexes (MI) and by the ratios between binucleated and mononucleated (B/M) cells. PMID- 14757194 TI - Two regions in chromosome 19q13.2-3 are associated with risk of lung cancer. AB - Lung cancer is the most common fatal cancer among Danish men, and the incidence rate is increasing among women. In a case-cohort study, we have investigated the occurrence of lung cancer in relation to a high-risk haplotype, previously identified for breast cancer among post-menopausal women, and in relation to the closely linked polymorphisms XPD Asp312Asn and Lys751Gln. Among 54220 members of a Danish prospective cohort study aged 50-64 at entry, 265 lung cancer cases were identified and a sub-cohort comprising 272 individuals was used for comparison. Among women in the 50-55 year age interval, homozygous carriers of the high-risk haplotype were at increased risk of lung cancer (RR=7.02, 95% CI=1.88-26.18). In the 56-60 year and 61-70 year age intervals, no associations were observed. Among men, no statistically significant associations were found in any age interval. Female homozygous carriers of the variant allele of XPD Lys751Gln were at significantly increased risk of lung cancer in the two younger age-intervals (50 55 years: RR=5.60, 95% CI=1.18-26.45, 56-60 years: RR=10.60, 95% CI=1.50-75.64). Among men, carriers of the variant allele of XPD Lys751Gln had a non significantly increased risk of lung cancer in the youngest age interval (RR=6.38, 95% CI=0.74-54.90). When the polymorphisms in XPD Asp312Asn and Lys751Gln were mutually adjusted, XPD Asp312Asn was not associated with increased risk of cancer. We found no interaction between genotypes and duration of smoking. In conclusion, two regions of chromosome 19q13.2-3 seem to be associated with risk of lung cancer. PMID- 14757195 TI - Identification of paraoxonase 3 gene (PON3) missense mutations in a population of southern Italy. AB - PON gene family includes at least three members termed PON1, PON2 and PON3, and it is mapped on human chromosome 7q21-q22. PON1 and PON3 gene products are constituents of high density lipoprotein (HDL) and have many enzymatic properties and antioxidant activity. PONs are proposed to participate in the prevention of low density lipoprotein (LDL) oxidation. PON1 and PON2 genes have missense polymorphisms, but, to date, no missense variants are reported in PON3 gene. In this work we explored the existence of genetic variants within the PON3 coding sequences. Five point mutations were identified by direct sequencing of genomic DNA derived from 250 randomly selected DNA samples of 1143 blood donors living in southern Italy. Three were silent mutations, while two were missense mutations that give rise to amino acid substitutions at positions 311 (S>T) and 324 (G>D). The missense variations in the DNA of the 1143 samples had frequencies of 0.22% (5 out of 2286 alleles) for the S311T mutation, and 0.57% (13 out of 2286 alleles) for the G324D mutation. The effect of these variants on the metabolic activity of paraoxonase 3 remains to be further evaluated. PMID- 14757196 TI - Mutagenicity of cadmium in mammalian cells: implication of oxidative DNA damage. AB - Cadmium and cadmium compounds are well established human carcinogens and are ubiquitously present in the environment. The carcinogenic mechanism(s) of cadmium remains largely unknown since direct mutagenic effect is weak in bacterial and in standard mammalian cell mutation assays. In this study, we show that when evaluated using the human-hamster hybrid A(L) cell mutation assay in which both intragenic and multilocus deletions can readily be detected, CdCl(2) is a strong mutagen that induces predominantly large deletion mutations. Concurrent treatment of A(L) cells with the oxyradical scavenger dimethyl sulfoxide significantly reduced the number of cadmium-induced mutations. In contrast, pre-treatment of cells with buthionine sulfoximine that depletes intracellular glutathione, increased cytotoxicity and mutagenicity of cadmium. These results demonstrate that reactive oxygen species mediate cadmium induced mutations in A(L) cells. With laser scanning confocal microscopy and the fluorescent probe 5-(and-6) chloromethyl-2',7'-dichlorodihydrofluorescein diacetate, we demonstrated that cadmium induced a dose and time dependent formation of intracellular oxyradicals. Using immunoperoxidase staining coupled with a monoclonal antibody-specific for 8 OHdG adducts in DNA, we demonstrated that cadmium induced a dose dependent increase of 8-OHdG adducts, which accumulated with prolonged exposure. Furthermore, we showed that at low concentration, cadmium, attenuated removal of hydrogen peroxide induced 8-OHdG adducts. Thus, the carcinogenicity of cadmium can, in part, be explained by its mutagenic activity, which is mediated by reactive oxygen species induced DNA damage and by its interference with the repair of oxidative DNA damage. PMID- 14757197 TI - Genoprotective pathways. Part I. Extracellular signaling through G(s) protein coupled adenosine receptors prevents oxidative DNA damage. AB - Adenosine has been previously shown to be a cytoprotective paracrine released by injured cells. However, it is presently unknown whether extracellular adenosine can prevent DNA damage. We show here that the adenosine analog, 2-chloroadenosine (2CA), has a potent (IC(50)=2.04 x 10(-9)M) genoprotective effect in cells subsequently exposed to H(2)O(2). The genoprotective signaling is transduced through adenosine receptors of the A2a and A2b subtypes. Increasing [cAMP](i) by forskolin or by cell permeable 8-br-cAMP produced a similar effect, whereas inhibiting the cAMP-mediated pathway with H-89, or increasing [nitric oxide](i) or [cGMP](i) blocked 2CA effect. Proteasomal inhibitors had no impact on 2CA signaling, while lithium chloride, an inhibitor of glycogen synthase kinase 3beta, completely blocked 2CA-induced genoprotective effect. These data indicate for the first time that extracellular adenosine protects cells against imminent oxidative DNA damage and suggest that prophylactic activation of this pathway prior to genotoxic challenges might reduce mutation rates. PMID- 14757199 TI - The role of bacterial and non-bacterial toxins in the induction of changes in membrane transport: implications for diarrhea. AB - Bacterial toxins induce changes in membrane transport which underlie the loss of electrolyte homeostasis associated with diarrhea. Bacterial- and their secreted toxin-types which have been linked with diarrhea include: (a) Vibrio cholerae (cholera toxin, E1 Tor hemolysin and accessory cholera enterotoxin); (b) Escherichia coli (heat stable enterotoxin, heat-labile enterotoxin and colicins); (c) Shigella dysenteriae (shiga-toxin); (d) Clostridium perfringens (C. perfringens enterotoxin, alpha-toxin, beta-toxin and theta-toxin); (e) Clostridium difficile (toxins A and B); (f) Staphylococcus aureus (alpha haemolysin); (g) Bacillus cereus (cytotoxin K and haemolysin BL); and (h) Aeromonas hydrophila (aerolysin, heat labile cytotoxins and heat stable cytotoxins). The mechanisms of toxin-induced diarrhea include: (a) direct effects on ion transport in intestinal epithelial cells, i.e. direct toxin interaction with intrinsic ion channels in the membrane and (b) indirect interaction with ion transport in intestinal epithelial cells mediated by toxin binding to a membrane receptor. These effects consequently cause the release of second messengers, e.g. the release of adenosine 3',5'-cyclic monophosphate/guanosine 3',5' monophosphate, IP(3), Ca2+ and/or changes in second messengers that are the result of toxin-formed Ca2+ and K+ permeable channels, which increase Ca2+ flux and augment changes in Ca2+ homeostasis and cause depolarisation of the membrane potential. Consequently, many voltage-dependent ion transport systems, e.g. voltage-dependent Ca2+ influx, are affected. The toxin-formed ion channels may act as a pathway for loss of fluid and electrolytes. Although most of the diarrhea-causing toxins have been reported to act via cation and anion channel formation, the properties of these channels have not been well studied, and the available biophysical properties that are needed for the characterization of these channels are inadequate. PMID- 14757200 TI - Microcystin production by Radiocystis fernandoi (Chroococcales, Cyanobacteria) isolated from a drinking water reservoir in the city of Belem, PA, Brazilian Amazonia region. AB - During the monitoring of toxic cyanobacteria in the Utinga Reservoir, which is the main drinking water supply for the city of Belem, PA, Brazil, a Radiocystis fernandoi strain (SPC714) was isolated. This non-axenic strain was submitted to a toxicity bioassay with mice and microcystin production analyzed by HPLC-DAD. The species was identified based on cultured and natural preserved material. Morphometric, developmental and reproductive characteristics were analyzed. The strain was cultured in liquid ASM-1 medium, at 25+/-1 degrees C, at an incident irradiance of 20 micromol photon m(-2)s(-1) and constant aeration. At the end of the exponential growth phase, cells were lyophilized and submitted to toxicity tests. The strain showed high toxicity to mice, by intraperitoneal route, with an approximate LD100 of 60 mg kg(-1) of body weight, producing characteristic symptoms of hepatotoxicity. Analyses performed by HPLC-DAD confirmed the production of microcystins, in a concentration of 3.83 microg mg(-1) of lyophilized cells. This is the first reference related to the toxicity of the genus Radiocystis. PMID- 14757201 TI - cDNA sequence analysis of seven peptide toxins from the spider Selenocosmia huwena. AB - Seven cDNAs encoding six toxins HWTX-I, HWTX-II, HWTX-IIIa, HWTX-IV, HWTX-V, HWTX VII and one lectin SHL-I, from the spider Selenocosmia huwena, were cloned and sequenced. On the basis of their amino acid sequences, we designed and synthesized 3' RACE and 5' RACE primer. By overlapping the two partial cDNA sequences obtained by 3' and 5' RACE, their full-length cDNA sequences were obtained. All of the cDNAs of these seven peptides encode a precursor including a potential signal peptide of 21-24 residues, a mature toxin of about 30 residues and an intervening pro region. The prepro regions of HWTX-I, HWTX-IIIa, HWTX-IV, HWTX-V and SHL-I were demonstrated, by the comparison of the cDNA sequences, to have high similarity, which is concert with the similar inhibitor cystine knot motif of HWTX-I, HWTX-IV and SHL-I although their functions are different. It was also demonstrated that, HWTX-II and HWTX-VII share the highly similar prepro region which is different from that of HWTX-I, HWTX-IV and SHL-I. The three dimensional structure of HWTX-II has been determined to exhibit a different motif. This indicates that the seven peptides from S. huwena could be classified into two different superfamilies according to the prepro region of cDNA sequences. PMID- 14757202 TI - Production of monoclonal antibodies capable of neutralizing dermonecrotic activity of Loxosceles intermedia spider venom and their use in a specific immunometric assay. AB - We have produced 13 mAbs for Loxosceles intermedia crude venom. Twelve were reactive against proteins of 32-35 kDa and one of these Li mAb(7) showed high neutralizing potency for the dermonecrotic activity of L. intermedia venom. This Li mAb(7) showed no cross-reactivity, with Loxosceles laeta (Brazil), L. laeta (Peru) and Loxosceles gaucho venoms. The mAbs were produced by immunization with the crude venom and screened by enzyme-linked immunosorbent assay (ELISA) using L. intermedia whole venom or dermonecrotic fraction (DNF) as antigens coated onto microtitre plates. A sensitive two-site immunometric assay was designed and shown to be useful for identifying and quantifying DNF from L. intermedia in biological samples. The Li mAb(7) coated onto microtitre plates and hyperimmune horse anti L. intermedia IgGs prepared by immunoaffinity chromatography and conjugated to horseradish peroxidase were used to set up a sandwich-type ELISA. Measurable absorbance signals were obtained with 0.2 ng of L. intermedia crude venom per assay. PMID- 14757203 TI - Pathological effects on mice by gambierol, possibly one of the ciguatera toxins. AB - Gambierol was isolated from Gambierdiscus toxicus, which causes ciguatera fish poisoning. The acute toxicological effects induced in mice by synthesized gambierol were studied. The lethal doses were about 80 microg/kg by i.p. and i.v., and 150 microg/kg by p.o. The main injury by this toxin was observed in the lung, and secondary in the heart, resulting in systemic congestion. Another toxic effect was seen in the stomach, inducing hypersecretion and ulceration. With survival from the severe stage during the initial 3 h, recovery was favorable, especially after 4 days. Additional effects were not evident during 1-week post administration observation. PMID- 14757204 TI - Absence of myocardial involvement in children victims of Crotalus durissus terrificus envenoming. AB - The myotoxic activity of the venom of Crotalus durissus terrificus is demonstrable by increased serum levels of the enzymes creatine kinase (CK), lactate dehydrogenase (LD), and aspartate aminotransferase. Serial measurements of CK, LD and their isoenzymes in bite victims showed a pattern similar to that observed in acute myocardial infarction, although the clinical course and electro and echocardiographic data did not suggest cardiac involvement. These data have raised the hypothesis that crotalid venom preferentially causes damage to type I and/or type IIa fibers, which contain quantities of CK-MB and LD1 similar to those found in cardiac fibers. In order to detect a possible concomitant silent cardiac involvement, seven children with severe crotalid envenoming were studied. Serum troponin I, determined more than once in each patient, were found to be normal. These data demonstrate the absence of cardiac involvement in these patients envenomed by C. durissus terrificus and confirmed the skeletal muscle origin of the elevated CK-MB. PMID- 14757205 TI - Structure and chromosomal localization of the gene for crotamine, a toxin from the South American rattlesnake, Crotalus durissus terrificus. AB - Crotamine is a 42 amino acid-long basic polypeptide, one of the major components of the South American rattlesnake, Crotalus durissus terrificus, venom. The mRNA has about 340 nucleotides and codifies a pre-crotamine, including the signal peptide, the mature crotamine, and a final lysine. In this report, we describe the crotamine gene with 1.8 kb organized into three exons separated by a long phase-1 (900 bp) and a short phase-2 (140 bp) introns. Exon 1 includes the 5' untranslated region and codifies the first 19 amino acids of the signal peptide. Exon 2 codifies 42 amino acids, three belonging to the signal peptide and 39 to the mature crotamine. Exon 3 codifies the last three amino acids of the mature toxin and the terminal lysine. The crotamine gene was mapped by in situ hybridization to the end of the long arm of chromosome 2, the intensity of signals differing between the two homologues. This may reflect a difference in gene copy numbers between chromosomes, a possible explanation for the variable amounts of crotamine found in the venom. PMID- 14757206 TI - Molecular cloning, expression and characterization of three short chain alpha neurotoxins from the venom of sea snake--Hydrophiinae Hydrophis cyanocinctus Daudin. AB - Three different genes named sn311, sn316 and sn285 were discovered by large-scale randomly sequencing the high quality cDNA library of the venom glands from Hydrophiinae Hydrophis cyanocinctus Daudin. Sequence analysis showed that these three genes encoded three different short chain alpha-neurotoxins of 81 amino acids, which contained a signal peptide of 21 amino acids and followed by a mature peptide of 60 amino acids. Amino acid comparison reveals that mature peptides of sn311 and sn316 are highly homologous, with the only variance at position 46, which is Lys46 and Ser46, respectively. Whereas the mature peptide of sn285 lacks the most conserved amino acids in short chain alpha-neurotoxins, Asp31 and Arg33. The coding sequences of three neurotoxins were cloned into a thioredoxin (TRX) fusion expression vector (pTRX) and expressed as soluble recombinant fusion proteins in E. coli. After purification, approximately 10 mg/l recombinant proteins with the purity up to 95% were obtained. These three recombinant proteins are designated as rSN311, rSN316 and rSN285, they have a molecular weight of 6.963, 6.920 and 6.756 kDa, respectively, which are similar to those predicted from amino acid sequences. LD50 values of rSN311, rSN316 and rSN285 are 0.0827, 0.095, and 0.0647 mg/kg to mice, respectively. Studies on effects of these recombinant proteins on neuromuscular transmission were carried out, and results indicate that they all can produce prompt blockade of neuromuscular transmission, but display distinct biological activity characteristic individually. The results from UV-circular dichroism (CD) spectra indicate that they share similar secondary structure compared to other identified alpha-neurotoxins, and no significant structural differences in these recombinant proteins are observed. PMID- 14757207 TI - The antisnake venom activities of Parkia biglobosa (Mimosaceae) stem bark extract. AB - Snake bites in rural Nigeria are commonly treated with plant extracts. We have studied the ability of one such traditionally used plant (Parkia biglobosa; [Jacq.] Benth., Mimosaceae) to reduce the effects of two snake venoms (Naja nigricollis, and Echis ocellatus) in several experimental models. A water methanol extract of P. biglobosa stem bark significantly (p<0.001) protected the chick biventer cervicis (cbc) muscle preparation from N. nigricollis venom induced inhibition of neurally evoked twitches when it was added to the bath 3-5 min before or after the venom. The extract also reduced the loss of responses to acetylcholine (Ach), carbachol and KCl, which are normally blocked by N. nigricollis venom, and significantly reduced the contractures of the preparation induced by venom. P. biglobosa extract (75, 150 and 300 microg/ml) significantly (p<0.05) protected C2C12 murine muscle cells in culture against the cytotoxic effects of N. nigricollis and E. ocellatus venoms. The extract protected egg embryos exposed to lethal concentrations of E. ocellatus venom for more than 12 h and completely blocked the haemorrhagic activity of the venom at concentrations of 5 and 10 microg/1.5 microl. P. biglobosa extract (400 mg/kg) did not protect mice injected i.p. with 5 and 2.5 mg/kg of E. ocellatus and N. nigricollis venoms, respectively. It, however, protected 40% of the mice from death caused by E. ocellatus venom after the extract and venom were pre-incubated for 30 min before injecting the mixture. PMID- 14757208 TI - Trocarin, a blood coagulation factor Xa homologue from snake venom, causes inflammation and mitogenesis. AB - Trocarin, a Group D prothrombin activator from Tropidechis carinatus snake venom, has high sequence similarity to blood coagulation factor Xa (FXa). Both trocarin and FXa activate prothrombin to mature thrombin and have similar requirements for cofactors, such as factor Va, Ca2+ ions and phospholipids. In addition to its hemostatic functions, human FXa causes inflammation and induces mitogenesis in several cell types due to its interaction with effector protease receptor-1 (EPR 1). The inter-EGF domain region (L83FTKRL88) of FXa implicated in EPR-1-binding is distinctly different in trocarin (K83VLYQS88). Here we show that, interestingly, trocarin also causes edema in the mouse footpad; the inflammation, accompanied by a large purplish clot, is more persistent than the transient edema caused by FXa. Histological examination indicates significant differences between edema induced by FXa and trocarin. Moreover, trocarin-induced edema is not inhibited by a synthetic peptide based on the FXa-binding region of EPR-1, indicating that the inflammation is probably mediated by a mechanism independent of EPR-1-binding. Trocarin, like FXa, also has a mitogenic effect on bronchial smooth muscle cells mediated by an EPR-1-independent mechanism. Hence trocarin, being closely related to FXa, has similar non-hemostatic functions in mediating inflammation and mitogenesis, yet appears to act by distinctly different mechanisms. PMID- 14757209 TI - Kinetics of accumulation and transformation of paralytic shellfish toxins in the blue mussel Mytilus galloprovincialis. AB - Mussels (Mytilus galloprovincialis) were fed cultures of the Paralytic Shellfish Poisoning agent Alexandrium minutum (Strain AL1V) for a 15-day period and, for the next 12 days, they were fed the non-toxic species Tetraselmis suecica, in order to monitor the intoxication/detoxification process. The toxin content in the bivalve was checked daily throughout the experiment. During the time-course of the experiment, the toxin profile of the bivalves changed substantially, showing increasingly greater differences from the proportions found in the toxigenic dinoflagellate used as food. The main processes involved in the accumulation of toxins and in the variation of the toxic profiles were implemented in a series of numerical models and the usefulness of those models to describe the actual intoxication/detoxification kinetics was assessed. Models that did not include transformations between toxins were unable to describe the kinetics, even when different detoxification rates were allowed for the toxins involved. The models including epimerization and reduction provided a good description of the kinetics whether or not differential detoxification was allowed for the different toxins, suggesting that the differences in detoxification rates between the toxins are not an important factor in regulating the change of the toxic profile. The implementation of Michaelis-Menten kinetics to describe the two reductive transformations produced a model that had a poorer fit to the data observed than the model that included only a first order kinetics. This suggests that, it is very unlikely that any enzymatic reaction is involved in the reduction of the hydroxycarbamate (OH-GTXs) to carbamate (H-GTXs) gonyautoxins. PMID- 14757210 TI - Immunized camel sera and derived immunoglobulin subclasses neutralizing Androctonus australis hector scorpion toxins. AB - Scorpion envenoming is a real health problem. The only specific treatment is immunotherapy with antibodies from immunized horses. The severity of scorpion envenoming and the rapid diffusion of the toxins into the blood compartment require an improvement of the present antivenom therapy. In this study, we report successful immunization of dromedaries (Camelus dromedarius) against the small weakly antigenic neurotoxins of Androctonus australis hector scorpion. Camel immune sera was tested for its specific antigenic reactivity and neutralizing capacity against Aah toxic fraction and AahII toxin. We demonstrate that a substantial proportion of polyclonal heavy chain antibodies bind to Aah toxins and in particular to AahII, the most toxic one scorpion venom component. Furthermore, we show that both dromedary sera and heavy chain antibody subclasses are capable of neutralizing the toxicity of Aah toxins in mice. PMID- 14757211 TI - PnTx4-3, a new insect toxin from Phoneutria nigriventer venom elicits the glutamate uptake inhibition exhibited by PhTx4 toxic fraction. AB - Several pools of neurotoxic peptides obtained from fractionated Phoneutria nigriventer venom induce different toxicological effects. One of them, PhTx4, is highly toxic towards insects and displays only a slight toxicity when injected in mice. Also, this fraction contains a class of peptides that are able to inhibit glutamate uptake in preparations of mammalian central nervous systems (CNS). In this work a new toxin called PnTx4-3 was isolated from the PhTx4 fraction by reverse phase and anion exchange steps using high performance liquid chromatography (HPLC). Edman sequencing of PnTx4-3 revealed that it was a polypeptide of 48 amino acid residues, containing 10 cysteines cross-linked by five disulfide bridges. The molecular mass measured by ES-Q-TOF mass spectrometry was 5199.49+/-0.64 Da, which is very close to the calculated mass from amino acid sequence (5199.99 Da). This toxin induces immediate excitatory effects when injected intrathoracically in house flies and cockroaches. Intracerebroventricular injections of 30 microg of PnTx4-3 in mice resulted in no apparent signs of intoxication. In order to make an orthologous comparison, pharmacological characterisation were carried out in rat brain synaptosomes by using [3H]-L-glutamate, showed that the whole PhTx4 fraction as well as the pure toxins PnTx4-3, Tx4(6-1) and Tx4(5-5) obtained of this fraction, were able to inhibit the glutamate uptake in the micromolar concentration range. PnTx4-3 inhibits the glutamate uptake in a dose dependent manner, with an IC50 of approximately 1 microM. PnTx4-3 is highly homologous to the Tx4(6-1) and Tx4(5-5) toxins previously described from the same fraction. PMID- 14757212 TI - Snake venom metalloproteinases: structure/function relationships studies using monoclonal antibodies. AB - Snake Venom Metalloproteinases (SVMPs) are synthesized as zymogens and undergo proteolytic processing resulting in a variety of multifunctional proteins. Jararhagin is a P-III SVMP, isolated from the venom of Bothrops jararaca, comprising metalloproteinase, disintegrin-like and cysteine-rich domains. The catalytic domain is responsible for the hemorrhagic activity. The disintegrin like/cysteine-rich domains block alpha2beta1 integrin binding to collagen and apparently enhance the hemorrhagic activity of SVMPs. The relevance of disintegrin-like domain is described in this paper using a series of mouse anti jararhagin monoclonal antibodies (MAJar 1-7). MAJar 3 was the only antibody able to completely neutralize jararhagin hemorrhagic activity. Neutralization of catalytic activity was partial by incubation with MAJar 1. MAJars 1 and 3 efficiently neutralized jararhagin binding to collagen with IC50 of 330 and 8.4 nM, respectively. MAJars 1 and 3 recognized the C-terminal portion of the disintegrin domain, which is apparently in conformational proximity with the catalytic domain according to additivity tests. These data suggest that disintegrin-like domain epitopes are in close contact with catalytic site or functionally modulate the expression of hemorrhagic activity in SVMPs. PMID- 14757213 TI - Phylogenetic conservation of a snake venom metalloproteinase epitope recognized by a monoclonal antibody that neutralizes hemorrhagic activity. AB - Snake venom metalloproteinases (SVMPs) are present in large quantities in venoms of viper snakes and also in some elapids. Jararhagin is a representative of a P III multidomain hemorrhagic SVMP present in Bothrops jararaca venom. It is comprised of a catalytic, a disintegrin-like and a cysteine-rich domain. Seven anti-jararhagin monoclonal antibodies (MAJar 1-7) were produced, of which six reacted with the disintegrin domain. MAJar 3 recognized an epitope present at the C-terminal part of the disintegrin-like domain, and neutralized jararhagin induced hemorrhage. In this study, we evaluated the reactivity of these monoclonal antibodies with venoms from 27 species of snakes belonging to different families. MAJar 3 recognized most of the hemorrhagic venoms. By ELISA, MAJar 3 reacted strongly with venoms from Viperidae family and weakly with Colubridae and Elapidae venoms. This recognition pattern was due to bands between 50 and 80 kDa, corresponding to P-III SVMPs. This antibody preferentially neutralized the hemorrhage induced by venoms of Bothrops snakes. This fact suggests that the epitope recognized by MAJar 3 is present in other metalloproteinases throughout snake phylogeny. However, slight structural differences in the epitope may result in insufficient affinity for neutralization of biological activities. PMID- 14757214 TI - Modulation of KCNQ4 channel activity by changes in cell volume. AB - KCNQ4 channels expressed in HEK 293 cells are sensitive to cell volume changes, being activated by swelling and inhibited by shrinkage, respectively. The KCNQ4 channels contribute significantly to the regulatory volume decrease (RVD) process following cell swelling. Under isoosmotic conditions, the KCNQ4 channel activity is modulated by protein kinases A and C, G protein activation, and a reduction in the intracellular Ca2+ concentration, but these signalling pathways are not responsible for the increased channel activity during cell swelling. PMID- 14757215 TI - Important role of raft aggregation in the signaling events of cold-induced platelet activation. AB - When human platelets are chilled below 20 degrees C, they undergo cold-induced activation. We have previously shown that cold activation correlates with the main phospholipid phase transition (10-20 degrees C) and induces the formation of large raft aggregates. In addition, we found that the glycoprotein CD36 is selectively enriched within detergent-resistant membranes (DRMs) of cold activated platelets and is extremely sensitive to treatment with methyl-beta cyclodextrin (MbetaCD). Here, we further studied the partitioning of downstream signaling molecules within the DRMs. We found that the phospholipase Cgamma2 (PLCgamma2) and the protein tyrosine kinase Syk do not partition exclusively within the DRMs, but their distribution is perturbed by cholesterol extraction. In addition, PLCgamma2 activity increases in cold-activated cells compared to resting platelets and is entirely inhibited after treatment with MbetaCD. The Src family protein tyrosine kinases Src and Lyn preferentially partition within the DRMs and are profoundly affected by removal of cholesterol. These kinases are non redundant in cold-activation. CD36, active Lyn, along with inactive Src and PLCgamma2 co-localize in small raft complexes in resting platelets. Cold activation induces raft aggregation, resulting in changes in the activity of these proteins. These data suggest a crucial role of raft aggregation in the early events of cold-induced platelet activation. PMID- 14757216 TI - Bacillus subtilis alpha-amylase: interactions of a partially folded conformer with small unilamellar vesicles. AB - We studied the interactions between conformers of exocellular alpha-amylase and small unilamellar vesicles (SUV) composed of the major membrane lipids of Bacillus subtilis under physiological conditions of pH, temperature and ionic strength. Using fluorescence spectroscopy, surface plasmon resonance (SPR) and phase separation, we show that the native alpha-amylase has no affinity for the SUV, whereas a partially folded form, displaying structural properties in common with the competent state for secretion, binds to the vesicles (KA approximately 10(5) M(-1)). This association prevented its subsequent folding. The complex was destabilized in the presence of PrsA, a major peripheric lipoprotein of B. subtilis which displays a strong affinity for SUV (KA approximately 1.5x10(8) M( 1)). Vesicles coated with PrsA lost their ability to bind the partially folded conformer. The approach in vitro, in which our aim was to mimic the last stage of alpha-amylase translocation, indicates that PrsA possibly helps, in vivo, the secreted protein to acquire its native conformation by modulating the interaction between the latter and the lipid polar heads on the trans side of the cytoplasmic membrane. PMID- 14757217 TI - Possible mechanism of polycation liposome (PCL)-mediated gene transfer. AB - A novel gene transfer system utilizing polycation liposomes (PCLs), obtained by modifying liposomes with cetyl polyethylenimine (PEI), was previously developed (Gene Ther. 7 (2002) 1148). PCLs show notable transfection efficiency with low cytotoxicity. However, the mechanism of PCL-mediated gene transfer is still unclear. In this study, we examined the intracellular trafficking of PCL-DNA complexes by using HT1080 cells, fluorescent probe-labeled materials, and confocal laser scan microscopy. We found that the PCL-DNA complexes were taken up into cells by the endosomal pathway, since both cellular uptake of the complex and gene expression were blocked by wortmannin, an inhibitor of this pathway. We also observed that the plasmid DNA and cetyl PEI complex became detached from the PCL lipids and was preferentially transferred into the nucleus in the form of the complex, whereas the PCL lipids remained in the cytoplasmic area, possibly in the endosomes. In fact, nigericin, which dissipates the pH gradient across the endosomal membrane, inhibited the detachment of lipids from the PCL-DNA complex and subsequent gene expression. Taken together, our data indicate the following mechanism for gene transfer by PCLs: PCLs effectively transfer DNA to endosomes and release cetyl PEI-DNA complexes into the cytosol. Furthermore, cetyl PEI also contributes to gene entry into the nucleus. PMID- 14757218 TI - Effect of sialyl Lewis X-glycoliposomes on the inhibition of E-selectin-mediated tumour cell adhesion in vitro. AB - The aim of this study was to evaluate the potential of different types of sialyl Lewis X-conjugated liposomes as competitive inhibitors for tumour cell adhesion to endothelial E-selectin. Sterically stabilised liposomes with the sLeX ligand at the terminal end of the polyethyleneglycol (PEG) chain, as well as vesicles that had the ligand embedded within the PEG-layer, were compared to ligand bearing liposomes without sterical stabilisation. First, 14 different tumour cell lines were characterised for their expression of sialyl Lewis X and/or A. Tumour cell adhesion was characterised in three static assays in vitro using: (i) immobilised E-selectin, (ii) CHO cells, transfected to express E-selectin and (iii) human umbilical vein endothelial cells (HUVEC). Sterically stabilised liposomes with the ligand at the terminal end of the polyethylene chain were the most effective inhibitors in all three assays and inhibited the adhesion of HT29 colon- and Lewis lung (LL) carcinoma cells by about 60-80%. The binding was not affected by a PEG-coating of the liposomes. Sterical stabilisation, on the other hand, completely prevented macrophage uptake (J774 cell line) independently of the presence of the ligand, while plain liposomes were taken up in an amount of 5.4 nmol liposomal lipids/10(6) macrophages. PMID- 14757219 TI - Characterization and transfection properties of lipoplexes stabilized with novel exchangeable polyethylene glycol-lipid conjugates. AB - The positive charge of cationic-lipid/DNA complexes (lipoplexes) renders them highly susceptible to interactions with the biological milieu, leading to aggregation and destabilization, and rapid clearance from the blood circulation. In this study we synthesized and characterized a set of novel amphiphiles, based on N-methyl-4-alkylpyridinium chlorides (SAINTs), to which a PEG moiety is coupled. Plasmids were fully protected in lipoplexes prepared from cationic SAINT 2 lipid and stabilized with SAINT-PEGs. Our results demonstrate that SAINT-PEG stabilization is transient, and permits DNA to be released from these lipoplexes. The rate of SAINT-PEG transfer from lipoplexes to acceptor liposomes was determined by the nature of the lipid anchor. Increased hydrophobicity, by lengthening the alkyl chain, resulted in a decrease of the rate of DNA release from the lipoplexes. Chain unsaturation had the opposite effect. Similarly, the in vitro transfection potency of lipoplexes containing PEG-SAINT derivatives was sensitive to the length and (un)saturation of the alkyl chain. However, the internalization of SAINT-PEG stabilized lipoplexes is determined by their charge, rather than by the concentration of the polymer conjugate. Lipoplexes targeted to cell-surface epithelial glycoprotein 2, by means of a covalently coupled monoclonal antibody, were specifically internalized by cells expressing this antigen. PMID- 14757220 TI - Overcoming the toxicity of membrane peptide expression in bacteria by upstream insertion of Asp-Pro sequence. AB - Transmembrane (TM) peptides often induce toxic effects when expressed in bacteria, probably due to membrane destabilization. We report here that in the case of the TM domains of hepatitis C virus (HCV) E1 and E2 envelope proteins, which are both particularly toxic for the bacteria, the insertion of the Asp-Pro (DP) sequence dramatically reduced their toxicities and promoted their expressions when produced as glutathione S-transferase (GST) GST-DP-TM chimeras. Subcellular fractionation showed that these chimeras co-sediment with the membrane fraction and contain active GST that could be solubilized with a mild detergent. Surprisingly, immuno-gold electron microscopy clearly showed that such chimeras are not localized in the membrane but in the cytosol. We thus postulate that they likely form proteo-lipidic aggregates, which prevent the bacteria from toxicity by sequestering the TM part of the chimeras. The reduction of toxicity in the presence of the Asp-Pro sequence is possibly due to Asp's negative charge that probably disadvantages the binding of the TM peptides to the membrane. In addition, the structural features of Pro residue could promote the formation of chimera aggregates. PMID- 14757221 TI - Glutamate 87 is important for menaquinol binding in DmsC of the DMSO reductase (DmsABC) from Escherichia coli. AB - Escherichia coli dimethylsulfoxide (DMSO) reductase is a trimeric enzyme with a catalytic dimer (DmsAB) and an integral membrane anchor (DmsC). Using site directed mutagenesis, we examined six residues in the periplasmic loop between helices two and three, potentially involved in menaquinol binding in DmsC. Mutants were characterised for growth, enzyme expression and activity, and 2-n heptyl-4-hydroxoquinoline N-oxide (HOQNO) inhibitor binding. Mutations of leucine 66, glycine 67, arginine 71, phenylalanine 73 and serine 75 had no effect on menaquinol binding. Only a glutamate residue (E87) located in helix three was important for menaquinol binding. E87 was replaced with lysine, glutamine and aspartate. All three mutants were assembled into the membrane. Neither the lysine nor the glutamine mutant enzymes were able to support anaerobic growth on glycerol/DMSO minimal media or oxidise lapachol. The glutamine mutant bound the inhibitor with lower affinity compared to wild-type, whereas in the lysine mutant, binding was almost abolished. The aspartate mutant behaved as a wild-type enzyme. The data shows that E87 is important for menaquinol binding and oxidation and is likely to act as a proton acceptor in the menaquinol binding site. PMID- 14757223 TI - Stability and solubility of integral membrane proteins from photosynthetic bacteria solubilized in different detergents. AB - As a first step toward the establishment of practical guidelines for the search for crystallization conditions, stability and solubility were examined for integral membrane proteins from photosynthetic bacteria in the presence of different detergents. The results obtained from their stability provided practical information on the proper choice of detergent type in the preparation process and the subsequent crystallization experiment. In addition, the determination of a solubility diagram provided a practical method for quantifying the correct choice of detergent concentration and for setting up the suitable precipitant concentration in the crystallization experiment. PMID- 14757222 TI - Co-expression of mCysLT1 receptors and IK channels in Xenopus laevis oocytes elicits LTD4-stimulated IK current, independent of an increase in [Ca2+]i. AB - Addition of LTD4 (10 nM) to Xenopus laevis oocytes expressing the mCysLT1 receptor together with hBK or hIK channels resulted in the activation of both channels secondary to an LTD4-induced increase in [Ca2+]i. In addition, the hIK channel is activated by low concentrations of LTD4 (<0.1 nM), which did not result in any increase in [Ca2+]i. Even though activation of hIK by low concentrations of LTD4 was independent of an increase in [Ca2+]i, a certain "permissive" level of [Ca2+]i was required for its activation, since buffering of intracellular Ca2+ by EGTA completely abolished the response to LTD4. Neither hTBAK1 nor hTASK2 was activated following stimulations with LTD4 (0.1 and 100 nM). PMID- 14757224 TI - Ouabain-insensitive Na+-ATPase of proximal tubules is an effector for urodilatin and atrial natriuretic peptide. AB - In the present paper we studied the effect of urodilatin and atrial natriuretic peptide (ANP) on the proximal tubule Na+-ATPase and (Na+K+)ATPase activities. Urodilatin and ANP inhibit the Na+-ATPase activity but not the (Na+K+)ATPase activity. Maximal effect was observed at a concentration of 10(-11) M for both peptides. In this condition, the enzyme activity decreases from 10.8 +/- 1.6 (control) to 5.7 +/- 0.9 or 6.1 +/- 0.7 nmol Pi mg(-1) min(-1) in the presence of urodilatin or ANP, respectively. This effect was completely reversed by 10(-6) M LY83583, a guanylyl cyclase inhibitor, and mimicked by 10 nM cGMP. Furthermore, both ANP and urodilatin increase cGMP production by 33% and 49%, respectively. This is the first demonstration that it was shown that urodilatin and ANP directly modulate primary active sodium transport in the proximal tubule. The data obtained indicate that this effect is mediated by the activation of the NPR A/guanylate cyclase/cGMP pathway. PMID- 14757225 TI - Role of toxin activation on binding and pore formation activity of the Bacillus thuringiensis Cry3 toxins in membranes of Leptinotarsa decemlineata (Say). AB - Binding and pore formation constitute key steps in the mode of action of Bacillus thuringiensis delta-endotoxins. In this work, we present a comparative analysis of toxin-binding capacities of proteolytically processed Cry3A, Cry3B and Cry3C toxins to brush border membranes (BBMV) of the Colorado potato beetle Leptinotarsa decemlineata (CPB), a major potato coleopteran-insect pest. Competition experiments showed that the three Cry3 proteolytically activated toxins share a common binding site. Also heterologous competition experiments showed that Cry3Aa and Cry3Ca toxins have an extra binding site that is not shared with Cry3Ba toxin. The pore formation activity of the three different Cry3 toxins is analysed. High pore-formation activities were observed in Cry3 toxins obtained by proteolytical activation with CPB BBMV in contrast to toxins activated with either trypsin or chymotrypsin proteases. The pore-formation activity correlated with the formation of soluble oligomeric structures. Our data support that, similarly to the Cry1A toxins, the Cry3 oligomer is formed after receptor binding and before membrane insertion, forming a pre-pore structure that is insertion-competent. PMID- 14757226 TI - Loop X/XI, the largest cytoplasmic loop in the membrane-bound melibiose carrier of Escherichia coli, is a functional re-entrant loop. AB - The melibiose carrier of Escherichia coli is a membrane-bound sugar-cation cotransporter consisting of 12 transmembrane helices connected by cytoplasmic and periplasmic loops, with both N- and C-terminus on the cytoplasmic side. Using a functional cysteine-less carrier, cysteine was substituted individually for residues 347-378 that comprise the largest cytoplasmic loop X/XI. The majority of the cysteine mutants have good protein expression levels. The cysteine mutants were studied for their transport activities, and the inhibitory effects of two sulfhydryl reagents, PCMBS (7-A long) and BM (29-A long). Cysteine substitution resulted in substantial loss of transport in 12 mutants. While PCMBS caused significant inhibition in only two mutants, T373C and V376C, from the periplasmic side (in a substrate-protective manner), more extensive inhibition pattern was observed from the cytoplasmic side, in seven mutants: V353C, Y358C, V371C, Q372C, T373C, V376C and G378C, suggesting that these residues are along the sugar pathway in the aqueous channel, close to the cytoplasmic side. Furthermore, the inhibitory effect of BM on the inside-out vesicles of the above mutants was clearly less than that of PCMBS, suggesting channel space limitation to large molecules, consistent with those residues being inside the channel. Three second site revertants (A350C/F268L, A350C/I22S, and A350C/I22N) were selected. They may suggest proximities between loop X/XI and helices I and VIII, in agreement with a re-entrant loop structure. Self thiol cross-linkings of the cysteine mutants on loop X/XI failed to form dimers, suggesting that most of the loop is not surface exposed from cytoplasmic side. Together, these results strongly indicated a functional re-entrant loop mechanistically important in Na+-coupled transporters. PMID- 14757227 TI - Molecular dynamics of the cyclic lipodepsipeptides' action on model membranes: effects of syringopeptin22A, syringomycin E, and syringotoxin studied by EPR technique. AB - Interaction of pore-forming toxins, syringopeptin22A (SP22A), syringomycin E (SRE) and syringotoxin (ST), with model membranes were investigated. Liposomes were prepared from saturated phospholipids (DPPC or DMPC) or from binary mixtures of DPPC with varying amount of DOPC or cholesterol. The effects of the three toxins on the molecular order and dynamics of the lipids were studied using electron paramagnetic resonance (EPR) techniques. SP22A was the most-, SRE less-, and ST the least effective to increase the ordering and to decrease the rotational correlation time of the lipid molecules. The effects were more pronounced: (a) on small unilamellar vesicles (SUVs) than on multilamellar vesicles (MUVs); (b) on pure DPPC than on DPPC-cholesterol or DPPC-DOPC mixtures. Fluidity changes, determined from EPR spectra at different concentrations of the toxin, suggested the shell structure of the lipid molecules in pore formation. EPR spectra observed at different depth of the hydrocarbon chain of the lipid molecules implied an active role of the lipid molecules in the architecture of the pores created in the presence of the three toxins. Temperature dependence of the fluidity of the SUVs treated with toxins has shown an abrupt and irreversible change in the molecular dynamics of the lipid molecules at a temperature close to the pretransition, depending on the toxin species and the lipid composition. Coalescence and aggregation of the SUVs were proposed as the origin of this irreversible change. PMID- 14757228 TI - Stabilisation of mixed peptide/lipid complexes in selective antifungal hexapeptides. AB - The design of antimicrobial peptides could have benefited from structural studies of known peptides having specific activity against target microbes, but not toward other microorganisms. We have previously reported the identification of a series of peptides (PAF-series) active against certain postharvest fungal phytopathogens, and devoid of toxicity towards E. coli and S. cerevisiae [Lopez Garcia et al. Appl. Environ. Microbiol. 68 (2002) 2453]. The peptides inhibited the conidia germination and hyphal growth. Here, we present a comparative structural characterisation of selected PAF peptides obtained by single-amino acid replacement, which differ in biological activity. The peptides were characterised in solution using fluorescence and circular dichroism (CD) spectroscopies. Membrane and membrane mimetic-peptide interactions and the lipid bound structures were studied using fluorescence with the aid of extrinsic fluorescent probes that allowed the identification of mixed peptide/lipid complexes. A direct correlation was found between the capability of complex formation and antifungal activity. These studies provide a putative structural basis for the mechanism of action of selective antifungal peptides. PMID- 14757229 TI - Intracellular accumulation of L-Arg, kinetics of transport, and potassium leak conductance in oocytes from Xenopus laevis expressing hCAT-1, hCAT-2A, and hCAT 2B. AB - Cationic amino acid transporters play an important role in the intracellular supply of L-Arg and the generation of nitric oxide. Since the transport of L-Arg is voltage-dependent, we aimed at determining the intracellular L-Arg concentration and describing the transport of L-Arg in terms of Michaelis-Menten kinetics, taking into account membrane voltage. The human isoforms of the cationic amino acid transporters, hCAT-1, hCAT-2A, and hCAT-2B, were expressed in oocytes from Xenopus laevis and studied with the voltage clamp technique and in tracer experiments. We found that L-Arg was concentrated intracellularly by all hCAT isoforms and that influx and efflux, in the steady state of exchange, were nearly mirror images. Conductance measurements at symmetric concentrations of L Arg (inside/outside) allowed us to determine KM and Vmax. The empty transporter of hCAT-2B featured an unexpected potassium conductance, which was inhibited by L Arg. PMID- 14757230 TI - Different structural requirements for adenylate cyclase toxin interactions with erythrocyte and liposome membranes. AB - The bifunctional Bordetella adenylate cyclase toxin-hemolysin (ACT) penetrates target cell membranes, forms cation-selective channels and subverts cellular signaling by catalyzing uncontrolled conversion of ATP to cAMP. While primarily targeting phagocytes expressing the alphaMbeta2 integrin (CD11b/CD18), the toxin can also penetrate mammalian erythrocytes lacking the receptor and membrane endocytosis. We sought here to analyze the membrane interactions of ACT in a liposome model. Insertion of ACT into liposome membranes required calcium and caused leakage of entrapped fluorescent probes due to liposome disruption, as indicated by similar release kinetics for the approximately 398 Da FITC probe and its approximately 4400 Da dextran conjugate. However, the non-acylated proACT, which does not penetrate cellular membranes, exhibited higher capacity to bind and lyze liposomes than the mature toxin, showing that the fatty-acyl modification was not required for penetration of ACT into the lipid bilayer. Individual deletions within the channel-forming, acylation and repeat domains of ACT abolished its capacity to disrupt both liposomes and erythrocytes. In contrast to erythrocyte binding, however, the liposome binding was only lost upon a simultaneous deletion of both the channel-forming and acylation domains, suggesting that the acylation domain was also involved in liposome penetration of ACT. Moreover, substitutions of glutamates 509 and 516 by lysines, which strongly enhanced the channel-forming and hemolytic activity of ACT, did not affect its capacity to disrupt liposomes. This shows that the mechanism of ACT action in cellular membranes is not fully reproduced in liposome membranes. PMID- 14757231 TI - Fluorescent modified phosphatidylcholine floppase activity of reconstituted multidrug resistance-associated protein MRP1. AB - Multidrug resistance-associated protein (MRP1) may function as a floppase in human red blood cells to translocate phosphatidylserine and/or phosphatidylcholine from inner membrane leaflet to outer leaflet. Here we report that the purified and reconstituted MRP1 protein into asolectin proteoliposomes is mainly in an inside-out configuration and possesses the ability to flop a fluorescent labeled phosphatidylcholine (NBD-PC) from outer leaflet (protoplasmic) to inner leaflet (extracytoplasmic). The reconstituted MRP1 protein retains endogenous ATPase activity. ATP hydrolysis is required for the flopping since removal of ATP and/or Mg2+ inhibits the translocation of NBD-PC. Further evidence to support this conclusion is that the translocation of NBD-PC is inhibited by vanadate, which traps ATP hydrolysis product ADP in the nucleotide binding domains. In addition, the translocation of NBD-PC by proteoliposomes containing MRP1 protein is in a glutathione-dependent manner, similar to the process of translocating anticancer drugs such as daunorubicin. verapamil, vincristine, vinblastine, doxorubicin and oxidized glutathione partially inhibited the translocation of NBD-PC, whereas MK 571, an inhibitor of MRP1 protein, inhibited the translocation almost completely. Taken together, the purified and reconstituted MRP1 protein possesses the ability to flop NBD-PC from outer to inner leaflet of the proteoliposomes. PMID- 14757232 TI - The voltage-dependent nonselective cation current in human red blood cells studied by means of whole-cell and nystatin-perforated patch-clamp techniques. AB - Human red blood cells (RBC) can be studied by means of whole-cell and nystatin perforated patch-clamp techniques. In 85% of the whole-cell experiments (n=86) and 69% of the perforated-patch recordings (n=13), steps to positive potentials, from a holding potential of 0 mV, induced a slow-activating non-inactivating persistent outward current which reverted at about 0 mV. The current activation phase fitted well with a two-component exponential curve. Half-maximal conductance was reached at about 42 mV. Na+ and K+ carried this current, which was not affected by 20 nM charybdotoxin or 20 mM TEA, but was reduced following a partial substitution of extracellular Cl- by tartrate. This current has characteristics similar to the single-channel currents already described in RBC and may be involved in the rapid adaptations of these cells in the circulation. PMID- 14757233 TI - Metabolism and function of coenzyme Q. AB - Coenzyme Q (CoQ) is present in all cells and membranes and in addition to be a member of the mitochondrial respiratory chain it has also several other functions of great importance for the cellular metabolism. This review summarizes the findings available to day concerning CoQ distribution, biosynthesis, regulatory modifications and its participation in cellular metabolism. There are a number of indications that this lipid is not always functioning by its direct presence at the site of action but also using e.g. receptor expression modifications, signal transduction mechanisms and action through its metabolites. The biosynthesis of CoQ is studied in great detail in bacteria and yeast but only to a limited extent in animal tissues and therefore the informations available is restricted. However, it is known that the CoQ is compartmentalized in the cell with multiple sites of biosynthesis, breakdown and regulation which is the basis of functional specialization. Some regulatory mechanisms concerning amount and biosynthesis are established and nuclear transcription factors are partly identified in this process. Using appropriate ligands of nuclear receptors the biosynthetic rate can be increased in experimental system which raises the possibility of drug-induced upregulation of the lipid in deficiency. During aging and pathophysiological conditions the tissue concentration of CoQ is modified which influences cellular functions. In this case the extent of disturbances is dependent on the localization and the modified distribution of the lipid at cellular and membrane levels. PMID- 14757234 TI - Effects of various culture environments on expression of major outer membrane proteins from Porphyromonas gingivalis. AB - We examined the effects of various culture environments on major outer membrane proteins from Porphyromonas gingivalis ATCC 33277. Major outer membrane protein patterns on gel electrophoresis showed little difference over the culturable range of osmolarity and pH. With elevated temperature or prolonged culture, the intensities of the gingipain bands decreased; however, bands of RagA, RagB and the putative porins were relatively stable. Similar results were observed with several different culture media. Although the precise functions of RagA, RagB and the putative porins are unknown, these factors may be strongly related to the initiation and progression of adult periodontitis. PMID- 14757235 TI - Permeabilizing effects of sub-inhibitory concentrations of microcystin on the growth of Escherichia coli. AB - Microcystin, a cyanotoxin produced by Microcystis aeruginosa, lacks potent antibacterial activity. When tested in combination, in vitro, inhibitory values for a range of hydrophobic antibiotics were significantly reduced in the presence of at least 1/20 x the minimum inhibitory concentration of microcystin. The degree of inhibition was equivalent to that of a well-characterised permeabilizing agent, polymyxin B nonapeptide. The permeabilizing ability of sub inhibitory concentrations of microcystin to affect the envelope of Escherichia coli was demonstrated by a rapid and sustained reduction in absorbance values of lysozyme-treated cells and by enhanced uptake of crystal violet in microcystin treated cultures. Direct effects of appropriate concentrations of microcystin on the integrity of bacterial outer and inner membranes were measured by release of specific enzyme markers. Although the exact mechanism for permeabilizing E. coli with microcystin has not been elucidated, the effects were consistent with permeability changes to the enterobacterial outer membrane caused by polymyxin B nonapeptide. PMID- 14757236 TI - Molecular cloning of a new laccase from the edible straw mushroom Volvariella volvacea: possible involvement in fruit body development. AB - Cloning of a laccase-encoding cDNA from the edible straw mushroom, Volvariella volvacea, was performed using reverse transcription-polymerase chain reaction (RT PCR) and rapid amplification of cDNA ends. The cDNA of the putative laccase gene (lac4) consisted of 1689 bp, including an open reading frame encoding a 23-amino acid signal peptide at the N-terminal end and a 540-amino acid mature protein with a predicted molecular mass of 58173 Da and a pI value of 6.1. The 10 histidine residues and one cysteine residue required to co-ordinate the four copper atoms at the active site of the protein were all conserved. The amino acid sequence of V. volvacea lac4 has a high degree of identity with other basidiomycete laccases. Transcription of the laccase gene was analysed by RT-PCR and, unlike many other laccase genes, shown to be regulated independently of either copper or aromatic compounds under the test conditions. However, the laccase gene is strongly expressed during that part of the mushroom developmental cycle involving fruit body morphogenesis. PMID- 14757237 TI - Characterisation of Pythium paroecandrum and its antagonism towards Botrytis cinerea, the causative agent of grey mould disease of grape. AB - Pythium paroecandrum (B-30), an oomycete, was isolated from soil samples taken from a wheat field in Genlis in the Burgundy region of France and was found to check the growth and development of Botrytis cinerea, a serious grapevine pathogen. The oomycete is a fast-growing organism, living on vegetable debris, and can be recognised by its catenulate hyphal swellings, catenulate oogonia, and monoclinous antheridia. When grown together with B. cinerea, the causal agent of the grey mould disease of the grapevine, P. paroecandrum shows a pronounced antagonism and suppresses its growth and its aptitude to provoke the grey mould symptoms. Morphological features of this oomycete, its antagonism to B. cinerea, the sequences of the internal transcribed spacer region of its nuclear ribosomal DNA, and its comparison with related species are discussed in this article. PMID- 14757238 TI - Isolation and characterization of the tobramycin biosynthetic gene cluster from Streptomyces tenebrarius. AB - The biosynthetic gene cluster for tobramycin, a 2-deoxystreptamine-containing aminoglycoside antibiotic, was isolated from Streptomyces tenebrarius ATCC 17920. A genomic library of S. tenebrarius was constructed, and a cosmid, pST51, was isolated by the probes based on the core regions of 2-deoxy-scyllo-inosose (DOI) synthase, and L-glutamine:DOI aminotransferase and L-glutamine:scyllo-inosose aminotransferase. Sequencing of 33.9 kb revealed 24 open reading frames (ORFs) including putative tobramycin biosynthetic genes. We demonstrated that one of these ORFs, tbmA, encodes DOI synthase by in vitro enzyme assay of the purified protein. The catalytic residues of TbmA and dehydroquinate synthase were studied by homology modeling. The gene cluster found is likely to be involved in the biosynthesis of tobramycin. PMID- 14757239 TI - Susceptibility of methicillin-resistant staphylococci to oregano essential oil, carvacrol and thymol. AB - The aim of this study was to evaluate the susceptibility of methicillin susceptible and methicillin-resistant staphylococci (MSS, MRS) to oregano essential oil, carvacrol and thymol. The commercial aerial parts of Origanum vulgare L. were hydrodistilled and the essential oil analysed by gas- chromatography/electron impact mass spectrometry. The inhibition efficacy of this essence and its major components was assayed against 26 MSS and 21 MRS, using an agar dilution method. The methicillin resistance was thoroughly typed by Epsilometer test (E-test), polymerase chain reaction for mecA gene detection and PBP2' latex agglutination test. The results clearly demonstrated that the comparison between the susceptibility of MSS and MRS to oregano oil, carvacrol and thymol showed no significant differences (Fisher's exact test, P > 0.05). The best minimum inhibitory concentration values were reported for carvacrol (0.015 0.03%, v/v) followed by thymol (0.03-0.06%, v/v) and oregano oil (0.06-0.125%, v/v). PMID- 14757240 TI - Diversity of chromosomal AmpC beta-lactamases from Enterobacter cloacae isolates in a Portuguese hospital. AB - Six clinical isolates of Enterobacter cloacae isolated in a Portuguese hospital, between April 1999 and November 2000, demonstrated resistance to almost all broad spectrum cephalosporins, except to cefepime. These isolates were susceptible to quinolones and to aminoglycosides. Isoelectric focusing demonstrated production of beta-lactamases with pIs > 8.0 and by all six isolates, exhibiting a cephalosporinase phenotype. The results of pulsed field gel electrophoresis revealed that these isolates were genetically unrelated. The amino acid sequence of six AmpC beta-lactamases (Eclo1FF, Eclo6FF, Eclo9FF, Eclo10FF, Eclo11FF and Eclo15FF) shared 97-99% homology with the chromosomal AmpC beta-lactamase from E. cloacae P99 and 86-87% homology with those of two plasmid-mediated AmpC beta lactamases, MIR-1 and ACT-1. This is the first report of chromosomal AmpC beta lactamase production by E. cloacae isolates in a Portuguese hospital. PMID- 14757241 TI - Identification of novel virulence-associated loci in uropathogenic Escherichia coli by suppression subtractive hybridization. AB - To identify novel virulence-associated genes in uropathogenic Escherichia coli (UPEC) strains, a suppression subtractive hybridization strategy was applied to genomic DNA of four clinical UPEC isolates from patients suffering from cystitis or pyelonephritis. The genomic DNA of four isolates (tester strains) was subtracted from the DNA of two different driver strains, the well characterized UPEC strain CFT073 and the non-pathogenic E. coli K-12 strain MG1655. We determined the sequence of 172 tester strain-specific DNA fragments, 86 of which revealed only low or no homology to nucleotide sequences of public databases. We further determined the virulence association of the 86 novel DNA fragments using each DNA fragment as a probe in Southern hybridizations of a reference strain collection consisting of 60 extraintestinal pathogenic E. coli isolates, and 40 non-virulent E. coli strains from stool samples. From this, 19 novel DNA fragments were demonstrated to be significantly associated with virulent strains and thus may represent new virulence traits. Our results support the idea of a considerable genetic variability among UPEC strains and suggest that novel genomic determinants might contribute to virulence of UPEC. PMID- 14757242 TI - Characterization of alkaliphilic laccase activity in the culture supernatant of Myrothecium verrucaria 24G-4 in comparison with bilirubin oxidase. AB - An enzyme showing alkaliphilic laccase activity was purified from the culture supernatant of Myrothecium verrucaria 24G-4. The enzyme was highly stable under alkaline conditions, showed an optimum reaction pH of 9.0 for 4 aminoantipyrine/phenol coupling, and decolorized synthetic dyes under alkaline conditions. It showed structural and catalytic similarities with bilirubin oxidase, but preferably oxidized phenolic compounds. The enzyme catalyzed veratryl alcohol oxidation at pH 9.0 with 2,2'-azino-bis(3-ethylbenzothiazoline-6 sulfonic acid) as a mediator, suggesting that the laccase mediator system functioned well under alkaline conditions. PMID- 14757243 TI - In silico analysis of the sigma54-dependent enhancer-binding proteins in Pirellula species strain 1. AB - The planctomycetes are a phylogenetically distinct group of bacteria, widespread in aquatic and terrestrial environments. Their cell walls lack peptidoglycan and their compartmentalised cells undergo a yeast-like budding cell division process. Many bacteria regulate a subset of their genes by an enhancer-dependent mechanism involving the alternative sigma factor sigma54 (RpoN, sigmaN) in association with sigma54-dependent transcriptional activators known as enhancer-binding proteins (EBPs). The sigma54-dependent regulon has previously been studied in several groups of bacteria, but not in the planctomycetes. We wished to exploit the recently published complete genome sequence of Pirellula species strain 1 to predict and analyse the sigma54-dependent regulon in this interesting group of bacteria. The genome of Pirellula species strain 1 encodes one homologue of sigma54, and 16 sigma54-dependent EBPs, including 10 two-component response regulators and a homologue of Escherichia coli RtcR. Two EBPs contain forkhead associated domains, representing a novel protein domain combination not previously observed in bacterial EBPs and suggesting a novel link between the enhancer-dependent regulon and 'eukaryotic-like' protein phosphorylation in bacterial signal transduction. We identified several potential sigma54-dependent promoters upstream of genes and operons including two homologues of csrA, which encodes the global regulator CsrA, and rtcBA, encoding a RNA 3'-terminal phosphate cyclase. Phylogenetic analysis of EBP sequences from a wide range of bacterial taxa suggested that planctomycete EBPs fall into several distinct clades. Also the phylogeny of the sigma54 factors is broadly consistent with that of the host organisms. These results are consistent with a very ancient origin of sigma54 within the bacterial lineage. The repertoire of functions predicted to be under the control of the sigma54-dependent regulon in Pirellula shares some similarities (e.g. rtcBA) as well as exhibiting differences with that in other taxonomic groups of bacteria, reinforcing the evolutionarily dynamic nature of this regulon. PMID- 14757244 TI - The N-terminal domain of yeast Bap2 permease is phosphorylated dependently on the Npr1 kinase in response to starvation. AB - The Saccharomyces cerevisiae branched-chain amino acid permease Bap2p plays a major role in leucine, isoleucine, and valine transport, and its synthesis is regulated transcriptionally. Bap2p undergoes a starvation-induced degradation depending upon ubiquitination and the functions of N- and C-terminal domains of Bap2p. Here we show that the N-terminal domain of Bap2p is phosphorylated in response to rapamycin treatment when both the N- and C-termini of Bap2p are fused to glutathione S-transferase. The phosphorylation is dependent on Ser/Thr kinase Npr1p. In npr1 cells, Bap2p becomes slightly more susceptible to the rapamycin induced degradation, suggesting that Npr1p counteracts the degradation system for Bap2p. PMID- 14757245 TI - Specific detection of the toxigenic species Fusarium proliferatum and F. oxysporum from asparagus plants using primers based on calmodulin gene sequences. AB - Fusarium proliferatum and Fusarium oxysporum are the causal agents of a destructive disease of asparagus called Fusarium crown and root rot. F. proliferatum from asparagus produces fumonisin B1 and B2, which have been detected as natural contaminants in infected asparagus plants. Polymerase chain reaction (PCR) assays were developed for the rapid identification of F. proliferatum and F. oxysporum in asparagus plants. The primer pairs are based on calmodulin gene sequences. The PCR products from F. proliferatum and F. oxysporum were 526 and 534 bp long, respectively. The assays were successfully applied to identify both species from the vegetative part of the plants. PMID- 14757246 TI - The yjoB gene of Bacillus subtilis encodes a protein that is a novel member of the AAA family. AB - The yjoB gene of Bacillus subtilis encodes a 48.8-kDa protein belonging to the AAA family. Members of this family contain a 200-250-amino acid residues AAA domain carrying a Walker A and B ATP-binding site assumed to be part of a molecular chaperone. The yjoB gene belongs to the sigmaW regulon, and members of this regulon have been reported to be transiently induced when cells enter the stationary growth phase. This assumption was confirmed here for yjoB by Western blot experiments and by analysis of a transcriptional fusion. Purified YjoB protein exhibited ATPase activity but was unable to prevent aggregation of denatured citrate synthase. An alignment of YjoB with a subgroup of AAA proteins present in Archaea suggests that YjoB might be involved in the modulation of the activity of one or more proteases. PMID- 14757247 TI - Cloning and analysis of WF146 protease, a novel thermophilic subtilisin-like protease with four inserted surface loops. AB - Cloning and sequencing of the gene encoding WF146 protease, an extracellular subtilisin-like protease from the thermophile Bacillus sp. WF146, revealed that the WF146 protease was translated as a 416-amino acid precursor consisting of a putative 18-amino acid signal peptide, a 10-kDa N-terminal propeptide and a 32 kDa mature protease region. The mature WF146 protease shares a high degree of amino acid sequence identity with two psychrophilic subtilisins, S41 (68.2%) and S39 (65.4%), and a mesophilic subtilisin, SSII (67.1%). Significantly, these closely related proteases adapted to different temperatures all had four inserted surface loops not found in other subtilisins. However, unlike those of S41, S39 and SSII, the inserted loops of the WF146 protease possessed stabilizing features, such as the introduction of Pro residues into the loop regions. Interestingly, the WF146 protease contained five of the seven mutations previously found in a hyperstable variant of subtilisin S41 obtained by directed evolution. The proform of WF146 protease (pro-WF146 protease) was overexpressed in Escherichia coli in an inactive soluble form. After heat treatment, the 42-kDa pro-WF146 protease converted to a 32-kDa active mature form by processing the N terminal propeptide. The purified mature WF146 protease hydrolyzed casein with an optimum temperature of 85 degrees C, and lost activity with a half-life of 30 min at 80 degrees C in the presence of 10 mM CaCl2. PMID- 14757248 TI - Purification and characterization of the (1-->3)-beta-glucanases from Acremonium sp. IMI 383068. AB - Three extracellular (1-->3)-beta-glucanases were purified from the fungus Acremonium sp. IMI 383068. Higher activities were unexpectedly obtained with pustulan, a (1-->6)-beta-glucan as carbon source, than when grown with laminarin, a (1-->3)-beta-glucan. Preliminary evidence suggests that these enzymes are not constitutive, but are inducible, and that their synthesis is repressed by glucose. All three had the same molecular masses, similar pH and temperature optima and none were glycosylated. They all appeared to have an exo-hydrolytic mode of substrate attack. N-terminal amino acid sequence data indicate that substantial post-translational modification of these had occurred, and that while two may be encoded by the same gene, the third may be genetically different. PMID- 14757249 TI - The activator of the Rhodospirillum rubrum PHB depolymerase is a polypeptide that is extremely resistant to high temperature (121 degrees C) and other physical or chemical stresses. AB - Hydrolysis of native (amorphous) polyhydroxybutyrate (nPHB) granules isolated from different sources by soluble PHB depolymerase of Rhodospirillum rubrum in vitro requires the presence of a heat-stable compound (activator). The activator was purified and was resistant against various physical and chemical stresses such as heat (up to 130 degrees C), pH 1-12, dryness, oxidation by H2O2, reducing and denaturing compounds (2-mercaptoethanol, 5 M guanidinium-HCl) and many solvents including phenol/chloroform. The activator coding gene was identified by N-terminal sequencing of the purified protein, and the deduced protein showed significant homology to magnetosome-associated protein (Mms16) of magnetotactic bacteria. Analysis of the activation process in vitro showed that the activator acts on nPHB granules but not on the depolymerase. The effect of the activator could be mimicked by pretreatment of nPHB granules with trypsin or other proteases but protease activity of the purified activator was not detected. Evidence is shown that different mechanisms were responsible for activation of nPHB by trypsin and activator, respectively. PHB granule-associated protein (PhaP) of Ralstonia eutropha nPHB granules were cleaved by trypsin but no cleavage occurred after activator treatment. Hydrolysis of artificial protein free PHB granules coated with negatively charged detergents (sodium dodecyl sulfate (SDS), cholate but not cetyltrimethyl-ammonium bromide (CTAB)) did not require activation and confirmed that surface layer proteins of nPHB granules are the targets of the activator rather than lipids. All experimental data are in agreement with the assumption that trypsin and the activator enable the PHB depolymerase to find and to bind to the polymer surface: trypsin by removing a portion of proteins from the polymer surface, the activator by modifying the surface structure in a not yet understood manner presumably by interaction with phasins of the proteinous surface layer of nPHB. PMID- 14757250 TI - Characterization of the protein processing and secretion pathways in a comprehensive set of expressed sequence tags from Trichoderma reesei. AB - Trichoderma reesei is a filamentous fungus widely used as an efficient protein producer and known to secrete large quantities of biomass degrading enzymes. Much work has been done aimed at improving the secretion efficiency of this fungus. It is generally accepted that the major bottlenecks in secretion are protein folding and ornamentation steps in this pathway. In an attempt to identify genes involved in these steps, the 5' ends of 21888 cDNA clones were sequenced from which a unique set of over 5000 were also 3' sequenced. Using annotation tools Gene Ontology terms were assigned to 2732 of the sequences. Homologs to the majority of Aspergillus niger's Srg genes as well as a number of homologs to genes involved in protein folding and ornamentation pathways were identified. PMID- 14757251 TI - Colony aggregation and biofilm formation in xylem chemistry-based media for Xylella fastidiosa. AB - Two chemically defined media based on xylem fluid chemistry were developed for Xylella fastidiosa. These media were tested and compared to chemically defined media XDM2, XDM4 and XF-26. New media were evaluated for the Pierce's disease (PD) strain UCLA-PD. Our media either was similar to the concentration of some amino acids found in the xylem fluid of the PD-susceptible Vitis vinifera cv. Chardonnay (medium CHARD2) or incorporated the tripeptide glutathione found in xylem fluid composition (medium 3G10-R). CHARD2 and 3G10-R are among the simplest chemically defined media available. Xylem fluid chemistry-based media supported X. fastidiosa growth and especially stimulated aggregation and biofilm formation. PMID- 14757253 TI - Many are called, but few are chosen. Feature selection and error estimation in high dimensional spaces. AB - We address the problems of feature selection and error estimation when the number of possible feature candidates is large and the number of training samples is limited. A Monte Carlo study has been performed to illustrate the problems when using stepwise feature selection and discriminant analysis. The simulations demonstrate that in order to find the correct features, the necessary ratio of number of training samples to feature candidates is not a constant. It depends on the number of feature candidates, training samples and the Mahalanobis distance between the classes. Moreover, the leave-one-out error estimate may be a highly biased error estimate when feature selection is performed on the same data as the error estimation. It may even indicate complete separation of the classes, while no real difference between the classes exists. However, if feature selection and leave-one-out error estimation are performed in one process, an unbiased error estimate is achieved, but with high variance. The holdout error estimate gives a reliable estimate with low variance, depending on the size of the test set. PMID- 14757254 TI - Parallel programming of gradient-based iterative image reconstruction schemes for optical tomography. AB - Optical tomography (OT) is a fast developing novel imaging modality that uses near-infrared (NIR) light to obtain cross-sectional views of optical properties inside the human body. A major challenge remains the time-consuming, computational-intensive image reconstruction problem that converts NIR transmission measurements into cross-sectional images. To increase the speed of iterative image reconstruction schemes that are commonly applied for OT, we have developed and implemented several parallel algorithms on a cluster of workstations. Static process distribution as well as dynamic load balancing schemes suitable for heterogeneous clusters and varying machine performances are introduced and tested. The resulting algorithms are shown to accelerate the reconstruction process to various degrees, substantially reducing the computation times for clinically relevant problems. PMID- 14757255 TI - Automatic evaluation of the image quality of a mammographic phantom. AB - In this work a method has been developed to analyse the digital image quality of a mammographic phantom by means of automatic process techniques. The techniques used for the digital image treatment are standard techniques as the image thresholding to detect objects, the regional growing for pixels pooling and the morphological operator application to determine the objects shape and size, etc. This study allows the obtention of information about the phantom characteristics, that due to its small size and lowly contrast can be obtained very difficultly by direct observation. The final aim of this work is to obtain one or more parameters to characterize the reference phantom quality image in an objective way. These parameters will serve to compare images obtained at different mammographic centers and also, to study the temporal evolution of the image quality produced by determined mammographic equipment. PMID- 14757256 TI - A semiparametric bootstrap approach to correlated data analysis problems. AB - In this note, we outline a simple to use yet powerful bootstrap algorithm for handling correlated outcome variables in terms of either hypothesis testing or confidence intervals using only the marginal models. This new method can handle combinations of continuous and discrete data and can be used in conjunction with other covariates in a model. The procedure is based upon estimating the family wise error (FWE) rate and then making a Bonferroni-type correction. A simulation study illustrates the accuracy of the algorithm over a variety of correlation structures. PMID- 14757257 TI - Quasi-automatic 3D finite element model generation for individual single-rooted teeth and periodontal ligament. AB - The paper demonstrates how to generate an individual 3D volume model of a human single-rooted tooth using an automatic workflow. It can be implemented into finite element simulation. In several computational steps, computed tomography data of patients are used to obtain the global coordinates of the tooth's surface. First, the large number of geometric data is processed with several self developed algorithms for a significant reduction. The most important task is to keep geometrical information of the real tooth. The second main part includes the creation of the volume model for tooth and periodontal ligament (PDL). This is realized with a continuous free form surface of the tooth based on the remaining points. Generating such irregular objects for numerical use in biomechanical research normally requires enormous manual effort and time. The finite element mesh of the tooth, consisting of hexahedral elements, is composed of different materials: dentin, PDL and surrounding alveolar bone. It is capable of simulating tooth movement in a finite element analysis and may give valuable information for a clinical approach without the restrictions of tetrahedral elements. The mesh generator of FE software ANSYS executed the mesh process for hexahedral elements successfully. PMID- 14757258 TI - A Paradox-based data collection and management system for multi-center randomized clinical trials. AB - We have developed a Paradox-based data collection and management system for large scale multi-site randomized clinical trials. The system runs under Windows operating system and integrates Symantec pcAnywhere32 telecommunications software for data transmission and remote control sessions, PKZIP utility for the compression/decompression of transmitted data, and Stat/Transfer for exporting the centralized Paradox database for analyses. We initially developed this system for VA Cooperative Study #399 'The Effect of Antiarrhythmic Therapy in Maintaining Stability of Sinus Rhythm in Atrial Fibrillation', which collects over 1000 variables on 706 patients at 20 sites. Patient intake for this 5-year study began in March of 1998. We have also developed an enhanced version of this system, which is being used in the NIH-funded 'Glucosamine/Chondroitin Arthritis Intervention Trial (GAIT)' that collects over 1200 variables on 1588 patients at 13 sites. Patient intake for this 4-year study began in October of 2000. PMID- 14757259 TI - Computer-assisted assessment of depression and function in older primary care patients. AB - We wanted to test the psychometric reliability and validity of self-reported information on psychological and functional status gathered by computer in a sample of primary care outpatients. Persons aged 65 years and older visiting a primary care medical practice in Baltimore (n=240) were approached. Complete baseline data were obtained for 54 patients and 34 patients completed 1-week retest follow-up. Standard instruments were administered by computer and also given as paper and pencil tests. Test-retest reliability estimates were calculated and comparisons across mode of administration were made. Separately, an interviewer administered a questionnaire to gauge patient attitudes and feelings after using the computer. Most participants (72%) reported no previous computer use. Nevertheless, inter-method reliability of the GDS15 at baseline (0.719, n=47), intra-method reliability of the computer in time (0.797, n=31), inter-method reliability of the CESDR20 at baseline (0.740, n=53), and the correlation between the CESDR20 computer version at baseline and follow-up (0.849, n=34) were all excellent. The inter-method reliability of the CESDR20 at follow-up (0.615, n=37) was lower but still acceptable. Although 28% were anxious prior to using the computer testing system, that percent decreased to 19% while using the system. The efficiency and reliability in comparison to the paper instruments were good or better. Even though most participants had not ever used a computer prior to participating in the study, they had generally favorable attitudes toward the use of computers, and also reported having favorable experience with the computer testing system. PMID- 14757260 TI - Antenatal olfactory learning influences infant feeding. AB - The aim is to know whether antenatal olfactory learning have a greater effect than postnatal olfactory learning on infant feeding even in the absence of triggering signals. We evaluated the sucking behavior of infants completely separated from their mothers for 10-14 days since birth. The 12 infants admitted to Chiba Children's Hospital were studied at 10-14 days of age. Oral feeding was initiated at 4-7 days of age. The sucking and expression pressures, frequency, and sucking efficiency were measured during bottle-feeding with exposure to odors of mother's milk, formula, and water. The mother's milk odor elicited more frequent sucking with higher expression pressure than did formula or water. In conclusion, the odor preferences acquired independently from postnatal experience may have a greater effect than postnatal olfactory learning on sucking activity. PMID- 14757261 TI - Diastolic left ventricular function in preterm infants with a patent ductus arteriosus: a serial Doppler echocardiography study. AB - In very low birth weight neonates, a left-to-right shunt via persistent ductus arteriosus (PDA) may interact with diastolic left ventricular function, but specific changes of Doppler parameters have yet to be reported. In a serial transmitral Doppler study, we investigated the impact of a PDA on diastolic function parameters. Twenty-two patients with and without PDA were examined on day 3.8+/-1 and day 14+/-2 after birth. By the first examination, 13 out of 22 patients had a PDA; by the second examination, the number was still 8 out of 22. Peak early and atrial flow velocities (44.8+/-15 and 50.1+/-13 cm/s, respectively) were higher (p<0.05) for neonates with PDA compared to those with closed duct (30.9+/-6 and 34.2 cm/s, respectively). Isovolumic relaxation time (IVRT) was shorter in neonates with PDA (45+/-7 ms, N=21) compared to those with a closed duct (55.3+/-5 ms, N=23) (p<0.01). IVRT correlated inversely with cardiac index (R=-0.79, p<0.01). All observed changes reversed to the normal range after closure of the PDA. When premature infants with a PDA experience a preload challenge, early and atrial peak velocities increase and IVRT shortens significantly. This coincidence of elevated transvalvular pressure differences and decreased IVRT in neonates with immature diastolic function can best be explained as a result of left atrial pressure elevation. Consequently, pulmonary venous pressure must be elevated, with its inherent effect on pulmonary capillary physiology. Thus, the monitoring of left ventricular diastolic function adds significant information to the care of preterm infants with a PDA. PMID- 14757262 TI - Doppler-derived parameters of diastolic left ventricular function in preterm infants with a birth weight <1500 g: reference values and differences to term infants. AB - Transmitral flow parameters in preterm and term infants were compared in order to study differences in signal expression and temporal dynamics of left ventricular diastolic function. In 63 preterm infants between 26 and 33 weeks of gestation and 102 term infants, a Doppler survey was performed during 6 months after birth. Early and atrial filling-time velocity integrals and peak velocities were significantly lower in the preterm neonates. Atrial filling parameters reached the level observed in term infants by 2 months of age. Peak early filling velocity was still lower for 2-month-old preterms and attained the term infants' level by 3 months of age. Preterm infants continued having high atrial filling fraction (AFF) (0.51+/-0.07) during 2 months after birth, while in term infants the fraction decreased continuously from 0.41+/-0.06 to 0.37+/-0.05. Isovolumic relaxation time (IVRT) was the only parameter without differences between preterm and term infants, and it decreased from 54+/-7 ms in neonates to 41+/-4 ms over 3 months. Stroke volume passing the mitral valve doubled in preterm (4+/-1 to 7.9+/ 1.5 ml/cm2), but increased by only 37% (6.9+/-1.6 to 9.5+/-2.2 ml/cm2) in term infants. Our observations show that the maturational period of diastolic function appears prolonged in preterm infants. As preterm infants have to cope with a higher physiologic preload augmentation during growth, part of the delay in parameter changes might be caused by preload stress rather than by persistence of functional impairment. Although doing well under physiological conditions, preterm neonates may be at higher risk for diastolic dysfunction than term infants when an additional preload challenge is encountered. PMID- 14757263 TI - Effects of different socioeconomic conditions on menarche in Turkish female students. AB - PURPOSE: The aim of this study was to determine the age at menarche, the menarcheal features, and the association between menarcheal age and socioeconomic status in an urban area in Turkey. In addition, we tried to assess whether there is a relationship between age at menarche and body composition. METHODS: We asked some questions about menarche of 1017 female adolescent students in the high schools of Manisa region. Height and weight were measured. The body mass index (BMI; kg/m2) was used as an index of relative weight. Adolescent girls were grouped into three socioeconomic status according to the educational and occupational levels of their parents. The age at menarche and the menarcheal pattern were evaluated according to the socioeconomic status. RESULTS: The ages of girls involved in the study ranged between 14 and 18 years, with a mean of 15.7+/-1.1 years. Although the menarcheal age was found to be lower in girls with higher socioeconomic status, there was no significant difference between the three different socioeconomic status. In all of the three groups, menarche was more common in summer and fall than in spring and winter. Although the mother was an important source of knowledge in all groups, it was significantly more important in the group with high socioeconomic status. Adolescent girls with low socioeconomic status had fewer premenstrual complaints. However, there was no significant difference between the groups. We found an inverse correlation between menarcheal age and postmenarcheal weight and the BMI (r=-0.14, p=0.000). However, there was no correlation between menarcheal age and postmenarcheal height. CONCLUSION: These results indicate that as the social status differences decrease, the difference observed in menarcheal age and pattern disappears in urban areas of developing countries. Menarcheal age may be an indicator of socioeconomic development. It does not influence postmenarcheal height; however, as menarcheal age decreases, BMI increases. PMID- 14757264 TI - Analyses of the potential oxygen transfer capability in placentae from infants succumbing to sudden infant death syndrome. AB - BACKGROUND: Morphometric oxygen diffusive conductance (Dp) was estimated to assess the potential efficiency of oxygen transfer across the materno-fetal interface in placentae obtained from victims of sudden infant death syndrome (SIDS). STUDY DESIGN: SIDS placentae were retrieved from archived storage and classified into normal birth weight (NBW, n=16), or small for gestational age (SGA, n=9) and compared against control placentae (n=40) or SGA (n=24) placentae. A combination of stereological techniques and physiological constants were used to estimate total Dp. RESULTS: SIDS NBW cases showed a crucial reduction in fetal capillary surface area when compared with control placentae. SIDS SGA showed a number of deficiencies in basic volumetric and surface area parameters. Values for total and specific Dp in placentae in both SIDS groups were maintained at levels comparable with control and SGA cases, respectively. CONCLUSION: Since more reductions were observed in SIDS SGA group, this suggests that factors responsible for these reductions maybe associated with SGA rather than being SIDS specific factors. PMID- 14757265 TI - Sweet babies: chocolate consumption during pregnancy and infant temperament at six months. AB - BACKGROUND: Chocolate contains several biologically active components potentially having behavioral and psychological consequences. AIMS: We tested whether chocolate consumption and stress experiences during pregnancy predict mother rated infant temperament at 6 months. DESIGN AND SUBJECTS: Prenatal frequency of chocolate consumption and intensity of psychological stress experience of the mothers, and temperament characteristics of the infants 6 months postpartum were evaluated in 305 consecutive, healthy mother-infant dyads. RESULTS: Mothers who reported daily consumption of chocolate rated more positively the temperament of their infants at 6 months. Maternal prenatal stress predicted more negatively tuned ratings of the infant temperament, particularly among those who reported never/seldom chocolate consumption. However, this effect was not observed among the mothers reporting weekly or daily chocolate consumption. CONCLUSIONS: In addition to producing subjective feelings of psychological well being, chocolate may have effects at multiple environmental and psychological levels. PMID- 14757266 TI - Changes in bone turnover during tibolone treatment. AB - OBJECTIVES: An open study was carried out to evaluate changes in bone remodeling markers such as N-telopeptide (NTx), tartrate-resistant acid phosphatase (TRAP), total alkaline phosphatase (TAP), and bone alkaline phosphatase (BAP) during a 1 year continuous tibolone treatment in postmenopausal women. MATERIAL AND METHODS: Thirty-six postmenopausal women were recruited for receiving tibolone 2.5 mg per day for 1 year. Densitometry and determination of biochemical markers of bone metabolism in serum and urine were performed at 1, 3, 6, and 12 months. RESULTS: Comparing baseline with 12 month's values, BAP and all resorption markers decreased significantly. NTx began to decrease since the initiation of the treatment (baseline: 74.4 +/- 5.3; 1 month: 57.5 +/- 4.2; 12 months: 36.6 +/- 2.8). BAP increased at the first month (baseline: 37.3 +/- 2.1; 1 month: 42.6 +/- 3.0) but diminished in the following months (12 months: 23.1 +/- 1.5). TAP started to decrease significantly only after 6 months of treatment (baseline: 37.3 +/- 2.1; 12 months: 31.4 +/- 2.3) and TRAP after 3 months (baseline: 9.8 +/- 0.4; 6 months: 9.1 +/- 0.5; 12 months: 8.2 +/- 0.4). Normal bone mineral density at distal and ultradistal forearm was maintained during the 1-year treatment (baseline: 0.42 +/- 0.01; 12 months: 0.42 +/- 0.01 and baseline: 0.33 +/- 0.01; 12 months: 0.33 +/- 0.01, respectively). CONCLUSION: The use of tibolone 2.5 mg per day diminished progressively and significantly bone resorption and formation markers during 1-year treatment period. PMID- 14757268 TI - Relationship between soft tissue body composition and bone mass in perimenopausal women. AB - OBJECTIVES: Perimenopause, the transition into menopause, marks the beginning of accelerated bone loss, contributing to the development of osteoporosis, a major public health problem. This perimenopausal transition has also been associated with a decrease in body lean mass, an increase in fat mass, and an increase in body weight. How these changes in fat mass and lean mass may influence bone mineral density (BMD) is currently unknown. The purpose of this study is to determine the independent effect and relative contribution of lean mass and fat mass to BMD in perimenopausal women. MATERIAL AND METHODS: The sample consisted of 43 sedentary perimenopausal women (age: mean = 49.6; S.D. = 3.2) with an intact uterus and ovaries, participating in a study of exercise and perimenopausal symptoms. Total body BMD, regional BMD, and soft tissue body composition were measured by dual-energy X-ray absorptiometry. Other measures including age, height, weight, and serum FSH and E2 were also obtained. RESULTS: Findings revealed that 14% of these perimenopausal women had low bone mass (osteopenia) in the lumbar spine and/or the femoral neck. Overall body fat mass and lean mass had positive relationships with BMD of lumber spine and the femur. However, using multiple regression analyses, only lean mass and ethnicity remained significant predictors for BMD of the femoral neck (r2 = 45%) with lean mass explaining more variance than ethnicity. Lean mass was the sole predictor of total proximal femur BMD explaining 38% of the variance. Fat mass was not a significant predictor of BMD at any skeleton site. CONCLUSIONS: These findings suggest that body lean mass, not fat mass, is a significant contributor to femoral BMD in perimenopausal women. PMID- 14757267 TI - Menopause in Croatia. Socio-demographic characteristics, women's attitudes and source of information, compliance with HRT. AB - OBJECTIVES: Hormone replacement therapy (HRT) in Croatian postmenopausal women, attitudes towards it and factors influencing this decision. METHODS: A total of 1100 women were interviewed in Ob/Gyn offices using self-response questionnaire of 56 items, dealing with demographic data, personal and family medical history, surgeries, menstrual and menopausal status, body mass index (BMI), sexuality, knowledge and attitudes towards the menopause and HRT. RESULTS: The adherence to HRT in Croatia was found to be very short, averaging only 3 months. Despite the differences between the users, non-users and past users, all groups give more reasons to take HRT, then reasons against it. Among the many demographic and patient history characteristics considered, the differences were only found regarding breastfeeding duration, menopausal age, use of oral contraceptives and IUDs. No differences were found in age, menarche, number of family members, number of deliveries, number of abortions, BMI, marital and employment status or level of education. Regarding personal and family medical history, an increased frequency was found in gallbladder and hepatic disorders for HRT users. The differences in current/past users ratio for different prescribers show that compliance with HRT is highest for university polyclinics and private gynaecological practices, and lowest for GPs and endocrinologists. CONCLUSION: The compliance with HRT in Croatia is very low, recommendations are mostly restricted to gynaecological practices and decision to take HRT is influenced mostly by psychological factors. PMID- 14757269 TI - Effect of postmenopausal hormone replacement therapy on cardiovascular performance. AB - Postmenopausal hormone replacement therapy (HRT) has usually been evaluated the relationship with atherosclerotic disease, whereas its effect on direct cardiac functions hasn't been investigated in detail. This study was planned to investigate the long-term effects of HRT on cardiac functions and exercise performance. METHODS: Thirty-six postmenopausal women (mean age: 51 +/- 4 years, 39-60 years) were prospectively analyzed with pulsed wave Doppler echocardiography and symptom-limited exercise stress test before HRT (oral 0.625 mg conjugated estrogen and 2.5 mg medroxyprogesteron acetate/day), and at the third and the sixth months. The effect of HRT on left ventricular ejection fraction (EF), early filling velocity (E wave) and late filling velocity (A wave), E wave deceleration time (EDT), E/A ratio, myocardial performance index (MPI), exercise duration and METS changes were examined. RESULTS: HRT did not significantly alter the left ventricular EF. At the third month of HRT, there was an insignificant increase in E wave, EDT, and E/A ratio, whereas an insignificant decrease was noted in MPI (P > 0.05). However, at the sixth month of HRT, these changes became significant (68 +/- 12 vs. 75 +/- 13 cm/s, P < 0.01; 171 +/- 24 vs. 184 +/- 14 ms, P < 0.01; 1.01 +/- 0.23 vs. 1.11 +/- 0.27, P < 0.01, and 44 +/ 9 vs. 39 +/- 8%, P < 0.001, respectively). On the other hand, exercise duration and exercise METS values showed significant improvements at the third month of HRT (423 +/- 104 vs. 482 +/- 104 s, P < 0.001; 8.2 +/- 1.7 vs. 9.1 +/- 2 METS, P < 0.001). These improvements also continued at the sixth month of HRT. In conclusion, postmenopausal HRT leads to a progressive improvement on left ventricular function parameters, and in parallel, in exercise performance. PMID- 14757270 TI - Fatty acid and cholesterol composition of the uterine artery intima in relation to menopausal status, age, and serum cholesterol. AB - OBJECTIVES: Estrogens modulate lipid metabolism and the increased risk of atherosclerosis in postmenopausal women is at least partly due to the reduction of estrogen production after menopause. We studied the effect of menopause on the contents of long-chain fatty acids, free cholesterol (FC) and cholesterol ester (CE) in uterine artery wall. METHODS: The uterine artery intima samples were obtained in connection with surgery of 21 postmenopausal and 51 premenopausal women. The amount of FC, CE and phospholipid fatty acids were measured by gas chromatography after extraction and fractionation and these lipid values were related to menopausal status, age and serum total and low-density lipoprotein (LDL) cholesterol levels. RESULTS: Premenopausal females had significantly less intimal FC (161 +/- 50 vs. 407 +/- 276 microg/100 mg wet weight, P = 0.003) and CE (19 +/- 34 vs. 305 +/- 348 microg/100 mg wet weight, P = 0.050) and smaller proportion of linoleic acid out of all phospholipid fatty acids (4.2 vs. 7.2%, P = 0.002) than postmenopausal women after adjustment with age. The content of CE (r = 0.34, P = 0.025) and the FC-to-CE ratio (r = -0.45, P = 0.002) correlated with age in premenopausal but not in postmenopausal women. Moreover, the intimal content of CE correlated with the percentage of intimal phospholipid linoleic acid in postmenopausal women (r = 0.79, P = 0.020). The same was true for FC (r = 0.73, P < 0.001). CONCLUSIONS: These results indicate that CE and FC accumulation into the wall of uterine artery depends on menopausal status, independently of age, and that the phospholipid long-chain fatty acid composition differs significantly between premenopausal and postmenopausal women. This suggests that estrogens may be involved in the regulation of artery wall lipid composition. PMID- 14757271 TI - Serum lipids in oophorectomized women during estrogen and testosterone replacement therapy. AB - OBJECTIVE: To evaluate the effects of giving testosterone undecanoate (TU) in addition to estrogen replacement on serum lipids in oophorectomized women. METHOD: Women with surgically induced menopause (n = 50) were randomly assigned to oral treatment with 2 mg of estradiol valerate in combination with 40 mg of TU or placebo for 24 weeks. The study was double-blind with cross-over to the other regimen for further 24 weeks of treatment. Forty-four women completed the study. Their serum concentrations of total, high density lipoprotein (HDL)- and low density lipoprotein (LDL)-cholesterol, triglycerides, lipoprotein-(a) (Lp-(a)), total testosterone, estradiol and sex hormone-binding globulin (SHBG) were analyzed at baseline and after 24 weeks of each treatment. RESULTS: Serum levels of total testosterone increased markedly from a baseline mean of 0.8-4.9 nmol/l during testosterone addition. The levels of free testosterone significantly increased during the combined treatment and fell when given estrogen alone. Total and LDL-cholesterol levels were significantly reduced by both treatments as also were those of Lp-(a) although the difference was not significant. We found a 13% reduction in HDL-cholesterol levels when testosterone was added, but no change with estrogen alone. Triglyceride levels were increased by estrogen treatment, but not affected by the combination of estrogen plus testosterone. CONCLUSIONS: These findings suggest that 40 mg of TU can be given in addition to estrogen replacement with only little side-effects on the pattern of circulating lipids. Although supraphysiological concentrations of testosterone were induced a significant reduction in total and LDL-cholesterol levels occurred. PMID- 14757272 TI - Menopause induced by oophorectomy reveals a role of ovarian estrogen on the maintenance of pressure homeostasis. AB - OBJECTIVES: Following spontaneous menopause women show a greater increase in systolic and diastolic blood pressure than men of the same age. The aim of the present study was to assess the effect of acute ovarian hormone withdrawal and replacement on blood pressure and forearm blood flow. METHODS: We studied 18 fertile middle-aged normotensive women (48 +/- 1.5 years, range 46-51 years) 1 week prior and 1 month subsequent to bilateral oophorectomy by means of 24-h blood pressure monitoring and strain-gauge venous occlusion plethysmography. Eighteen subjects who had undergone hysterectomy with ovarian sparing, matched for age and biophysical characteristics, were used as a control group. All women were free from cardiovascular risk factors or disease. RESULTS: Oophorectomy increased the mean values of 24 h (P < 0.001), daytime (P < 0.05), and nighttime (P < 0.01) diastolic blood pressure and nighttime systolic blood pressure (P < 0.01). Blood pressure increase was associated with a rise in forearm vascular resistance (P < 0.01). No significant changes in either blood pressure or forearm vascular resistance values were observed in hysterectomized women. In 16 oophorectomized women a 3-month estrogen replacement therapy (ERT) (17beta estradiol, 100 mcg/day by transdermal patches) brought blood pressure and forearm vascular resistance values to a level comparable to that recorded before intervention. CONCLUSIONS: Surgically-induced menopause causes an increase in peripheral vascular resistance and blood pressure suggesting a role of ovarian hormones in the homeostatic pressure modulation. Recovery of the baseline condition after ERT suggests that the accelerated increase in blood pressure after menopause is due to ovarian and above all estrogen insufficiency. PMID- 14757273 TI - Quantitative ultrasound (QUS) of bone in the management of postmenopausal women. AB - OBJECTIVES: Postmenopausal osteoporosis is becoming a major problem for healthcare institutions as it has a growing social and economic impact. The incidence of osteoporotic fractures is constantly increasing due to the increase in life expectancy. The gynaecologist plays an important role in establishing a "biological zero" in each perimenopausal patient, and controlling the rate of bone loss during postmenopausal period. RESULTS: Dual energy X-ray absorptiometry (DXA) has been widely used for the diagnosis and management of osteoporosis and represents a strong risk factor for fractures, but it presents several limitations with regards to diagnosis, treatment follow-up and differential diagnosis of secondary osteoporosis. In these last years quantitative ultrasound (QUS) technique has been introduced for the evaluation of bone status in postmenopausal women and several in vitro and clinical studies have demonstrated the reliability of the examination in terms of: reproducibility, evaluation of fracture risk, treatment follow-up, differential diagnosis. QUS has proven to be equally capable in the prediction of future osteoporosis related fractures in comparison to DXA. Large-scale cross-sectional and longitudinal studies have demonstrated the applicability of QUS in screening the female population during the climacteric period. QUS technique seems to be very efficient in identifying "fast losers", identifying subjects at risk for osteoporosis requiring second level investigation (DXA, X-ray), diagnosing secondary osteoporosis. CONCLUSION: If QUS is used in a systematic and rational manner in clinical practice, it is a valid technique for the prevention of osteoporosis in postmenopausal women. PMID- 14757274 TI - Menometrorrhagia in the perimenopause is associated with increased serum estradiol. AB - OBJECTIVE: The purpose of this study was to evaluate the possible association between menometrorrhagia and the level of endogenous estrogen in perimenopausal women. METHODS: A prospective controlled study in which 28 perimenopausal women > 40 years presenting with menometrorrhagia were compared with 28 age-matched (+/- 2 years) women with normal cyclical menstrual periods concerning levels of estradiol and follicle-stimulating hormone (FSH). Neither of the two groups had received sexual hormone treatment at least in 2 weeks preceding the hormonal assessment. RESULTS: The serum level estradiol in the patients was significantly higher than in the controls (0.55 nmol/l versus 0.24 nmol/l), whereas FSH was not significantly different between the two groups (21.2 IU/l versus 11.8 IU/l). Twenty of the 28 patients had performed at histologic examination of the endometrium, and 10 of these (50%) had signs of endometrial hyperplasia. No relationship was found between the endometrial histology and the estradiol level. CONCLUSIONS: An association between a high endogenous estradiol level and menometrorrhagia in the perimenopause was demonstrated. This may have implications for the choice of treatment in this group of women. It is proposed that this type of bleeding disturbances should be controlled by progestins only, and not with combined estrogen-progestin treatment. Suppression of the associated hyperestrogenism could be achieved by use of oral contraceptives or GnRH agonists. PMID- 14757276 TI - The role of p63 in development and differentiation of the epidermis. AB - Expression of p63, a transcription factor that is transcribed into six isoforms, is required for proper development of stratified epithelia, such as the epidermis. In the absence of p63, epithelia remain single-layered. The molecular role of p63 in development and differentiation of stratified epithelia, however, remains controversial. Based on recent studies, we now believe that p63 has a dual role and is essential for development as well as maintenance of the epidermis. During embryogenesis, p63 may be the molecular switch required for initiation of epithelial stratification. This is based on our recent data demonstrating that ectopic expression of a p63 isoform in single-layered epithelia results in the induction of a stratification program. Furthermore, in the mature epidermis, p63 may maintain the proliferative potential of basal keratinocytes. This is suggested by the observation that p63 is primarily expressed in the basal compartment of the epidermis, that p63 expression induces hyperproliferation, and that its expression needs to be downregulated for terminal differentiation to take place. In this review, we discuss recent evidence supporting this dual role for p63 and place it in the context of our increasing knowledge of epidermal development and differentiation. PMID- 14757277 TI - Comparative studies on level of androgens in hair and plasma with premature male pattern baldness. AB - BACKGROUND: It is well known that male-pattern baldness (MPB) is not started from occipital, but frontal or scalp of head. We can assume that distribution of androgenic steroids is different for each region of the head. OBJECTIVE: We hypothesize that the levels of androgenic steroids are different not only between vertex hair with MPB and controls but also between occipital hair with MPB and controls. Moreover, we want to search for the biochemical indicator in plasma and hair sample (baldness: 22, non-baldness: 13) obtained from dermatology of medical center. After then, we desire to present fundamental data regarding diagnosis, medical cure, and prevention for premature MPB. METHODS: After hair and plasma were hydrolyzed, and then extracted with organic solvent. To assess androgenic steroids levels, we used gas chromatography-mass spectrometry (GC-MS) system in selected ion monitoring mode. RESULTS: The level of dihydrotestosterone (DHT) and the ratio of testosterone to epitestosterone (T/E ratio) in vertex hair from premature baldness subjects were higher than in the sample of non-baldness subjects (P<0.001, 0.001), whereas the levels of androgens in occipital hair from the same baldness group were not different. In addition, we discovered the levels of DHT, testosterone, and DHT/T ratio in plasma from premature MPB were higher than in those of control subjects (P<0.001, 0.001, 0.005). CONCLUSION: We verified that the distribution of androgenic steroids is unlike in various regions of individual subjects. Moreover, the increased DHT/T ratio in balding plasma indirectly confirms the high activity of 5alpha-reductase type II. PMID- 14757278 TI - Expression of p53 family in scars. AB - BACKGROUND: There have been some reports on the relationship between p53 and keloid formation. However, there have been no studies comparing the p53 expression among scars in various stages of maturity. However, p63 and p73 have been identified as p53-related genes and have been found to be similar to p53 in their structures and functions and these proteins have also been suggested to relate to scar formation. OBJECTIVE: We investigate the expression of three proteins of the p53 family in scars with various clinical manifestations and discuss the shared features and differences of these proteins. METHODS: Forty untreated scar lesions consisting of keloids, hypertrophic scars, and atrophic scars were prepared for investigation. We detected the expression of p53, p63 and p73 proteins using Western blot analysis and histopathological study in each sample. RESULTS: The 40 lesions were divided into four groups according to their clinical manifestations: keloid (Group A), red hypertrophic scar (Group B), white and hard hypertrophic scar (Group C), atrophic white scar (Group D). In Groups A and B, the histopathological findings demonstrated increased fibroblasts, capillary vessels and infiltration of inflammatory cells. In Group C, most of these changes decreased but proliferation of collagen fibers was evident. In Group D, the degree of proliferation of collagen fibers was much less and capillary vessels and infiltration of inflammatory cells were not evident. The levels of p53 protein elevated in Groups A, B and C and were higher in order of Groups A, B and C. In Group D, the level of p53 was almost the same as that of the control. The level of p63 protein was almost the same as that of the control in all groups. The level of p73 protein was elevated only in Group C. CONCLUSION: The p53 family members behave in a different manner in various scar tissues. It is suggested that these proteins play different roles in scar formation and the development of unfavorable scars. PMID- 14757279 TI - Roxithromycin decreases ultraviolet B irradiation-induced reactive oxygen intermediates production and apoptosis of keratinocytes. AB - BACKGROUND: In addition to their antimicrobial action, roxithromycin (RXM), a new 14-membrane macrolide antibiotics, have immunomodulatory, anti-inflammatory and anti-oxidative activity. Ultraviolet B (UVB) irradiation induces reactive oxygen intermediates and apoptosis of keratinocytes. OBJECTIVE: To examine the anti apoptotic and anti-oxidative effect of RXM on UVB-irradiated keratinocytes. METHODS: UVB-induced apoptosis was determined by cell death assay using crystal violet staining, and DNA fragmentation assay. Superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione reductase (GR), and calatase activities were measured in UVB-irradiated SV40-trasnformed human keratinocytes (SVHK cells). Detection of superoxide was performed histologically using hydroethidine and colorimetric quantitative assay using ferrous irons. H(2)O(2) was measured by colorimetrical assay. RESULTS: RXM suppressed UVB-induced apoptosis of SVHK cells. UVB-irradiated SVHK cells showed decreased SOD, GPx, GR, and catalase activities. RXM pretreatment suppressed the decrease in these enzyme activities with the maximal effect detected at 10microM of RXM. The effect was associated with suppression of UVB-induced superoxide and H(2)O(2) production. CONCLUSION: The present study demonstrated that RXM has anti-apoptotic and anti-oxidative effects against UVB-irradiated keratinocytes. PMID- 14757280 TI - PCR and PCR-RFLP techniques targeting the DNA topoisomerase II gene for rapid clinical diagnosis of the etiologic agent of dermatophytosis. AB - BACKGROUND: We have focused on the DNA topoisomerase II genes of several pathogenic fungi, and developed polymerase chain reaction (PCR) and PCR restriction fragment length polymorphism (RFLP) methods targeting this gene for identification of dermatophytes. OBJECTIVE: To assess the availability of the PCR based identification for an etiologic study of dermatophytosis, by testing these PCR and PCR-RFLP methods for stability and reproducibility. METHODS: Three hundred and fifty-six dermatophyte strains were isolated from 305 patients with tinea, and their genomic DNAs were used as templates for the PCR using primer mixes (PsT, PsME, dPsD1 or dPsD2) composed of gene-specific primers for identification of dermatophytes to the species level. The genomic DNAs of Trichophyton rubrum were further subjected to subrepeat element analysis of the nontranscribed spacer (NTS) of ribosomal DNA (rDNA). RESULTS: In this study, six dermatophyte species (T. rubrum, Trichophyton mentagrophytes, Trichophyton tonsurans, Microsporum canis, Microsporum gypseum, and Epidermophyton floccosum) were obtained. In all cases, the identifications obtained from the PCR and PCR RFLP targeting the DNA topoisomerase II gene coincided with those from the conventional morphological features-based identification technique. The sensitivity of the PCR-based identification was found to be a colony of approximately 3mm in diameter. Furthermore, T. rubrum was divided into three groups (17 types) on the basis of the sizes and numbers of the products generated from the TRS-1 region, and three types from the TRS-2 region. CONCLUSION: The PCR and PCR-RFLP targeting the DNA topoisomerase II gene were rapid, stable, and reproducible for species identification of dermatophytes, and thus are convenient tools for an etiologic study of dermatophytosis. PMID- 14757281 TI - Expression of p53, bcl-2 and growth hormone receptor in atrophic type of actinic keratosis. PMID- 14757282 TI - Expression of phosphorylated Smad2 in normal human epidermis. PMID- 14757283 TI - NK-355, an aminovinyl photosensitizing dye, enhances collagen production through TGF-beta1 upregulation. PMID- 14757284 TI - Cholera toxin and retinoic acid slow the growth rate of human keratinocytes in low-calcium, serum-free culture. PMID- 14757285 TI - The Vitamin A derivative etretinate improves skin sclerosis in patients with systemic sclerosis. PMID- 14757288 TI - Strongest correlation of HbA(1c) with 1-month-earlier glucose levels in type 2 diabetes. AB - We examined the correlations of hemoglobin A(1c) (HbA(1c)) with each plasma glucose (PG) level obtained at 0 (the same day), 1 and 2 month(s) prior to HbA(1c) determination. Data were from glycemic profiles of four patients of type 2 diabetes mellitus treated with tablets whose HbA(1c) and pre- and post breakfast PG levels were monitored each month. There was no significant difference in the correlation coefficients in cases 1 and 2, who presented with linear glycemic time courses. In contrast, HbA(1c) correlated with 1-month earlier pre-breakfast PG level more strongly than 2-month-earlier post-breakfast PG level in cases 3 and 4, and than same-day post-breakfast PG level in case 3 (P<0.05, ANOVA). The cases 3 and 4 presented with fluctuating glycemic time courses. Samples were separated into upslope's and downslope's sections according to HbA(1c) fluctuation in the latter two cases. Reflecting around the 1-month lag between HbA(1c) and PG, the two sections' regression lines for PG versus HbA(1c) corresponded in the only samples related to 1-month-earlier pre- and post breakfast PG (t-test). In conclusion, it appears that pre- and post-breakfast PG levels are the most reliable predictors of 1-month-later HbA(1c) in type 2 diabetic outpatients who undergo medical examinations every month. PMID- 14757287 TI - Alteration in endothelin receptor sub-type responsiveness and in the endothelin TXA(2) mimetic U46619 interaction, in type-2 hypertensive diabetic Zucker rats. AB - BACKGROUND: Type-2 diabetes is characterized by endotheliopathy, which increases target organ damage and mortality. There is excessive endothelin-1 and TXA(2) production, and abnormal vascular reactivity to endothelin-1, manifested as a paradoxical hypotensive action in Zucker diabetic, but not lean rats. We examined the hypothesis that there is an alteration in the ET-A/ET-B receptor subtype sensitivity, and/or the interaction or cross-talk between ET-1 and TXA(2) in type 2 diabetes, using Zucker diabetic rats and their lean littermates. MATERIALS AND METHODS: Hemodynamic studies were performed in lean and Zucker fatty diabetic rats of both sexes. Laser doppler flowmetry was used to measure renal cortical (RCF) and medullary blood flow (MBF) responses. Dose response curves for mean arterial blood pressure (MAP), MBF and RCF in response to ET-1, U46619, acetylcholine, and L-NAME (25mg/kg) were constructed after pre-treatment of the rats with either BQ610 1mg/kg i.v. or BQ788 0.5mg/kg i.v. The effects of BQ610 and BQ788 on whole blood impedance aggregation were also assessed. RESULTS: BQ788, but not BQ610 abolished both the paradoxical hypotensive action of ET-1 in Zucker diabetic rats (n=7 each, P<0.001 ANOVA) as well as the dose-dependent rise in MBF (P<0.001 ANOVA). BQ788, but not BQ610 abolished the difference in response to ET-1 between lean and diabetic Zucker rats. U46619 caused a hypotensive action in male Zucker rats which was abolished by L-NAME 25mg/kg or indomethacin 10mg/kg i.v. The U46619 interaction with BQ788 on both MAP and MBF was significantly (P<0.03 ANOVA) different between lean and diabetic Zucker rats. BQ788, but not BQ610 attenuated both the MAP and MBF responses to acetylcholine or L-NAME P<0.02 ANOVA). However, BQ610 dose-dependently attenuated the slope of platelet aggregation in both lean and Zucker diabetic rats (P<0.02 ANOVA). CONCLUSION: ET B receptor antagonism abolished the abnormal vascular reactivity and MBF responses to ET-1, and also normalized the vasoactive responses to the level seen in healthy lean Zucker rats. ET-1 receptor blockade influences the responses to TXA(2) receptor activation. In the systemic and renal circulation, this interaction appears to be mostly ET-B receptor mediated, whilst in platelets, ET A receptor role may be predominant. The interaction or cross-talk between ET-1 and TXA(2) is altered in the type-2 diabetic state. Collectively, these pathophysiological changes may contribute to the vicious circle of diabetic endotheliopathy. PMID- 14757289 TI - In vitro mononuclear cell production of tumour necrosis factor-alpha and weight loss. AB - OBJECTIVE: Elevated tumour necrosis factor-alpha (TNF-alpha) production in adipose tissue has been associated with obesity. We investigated whether mononuclear cell production of TNF-alpha decreased with weight loss in an obese population. RESEARCH METHODS AND PROCEDURES: Seventeen obese patients with type 2 diabetes (BMI 32.5+/-0.9 kg/m(2)) and 33 obese, non-diabetic controls (BMI 31.2+/ 0.5 kg/m(2)) underwent 12 weeks of 30% total energy restriction (6622+/-84 KJ per day). Every 4 weeks, weight and blood pressure were measured and fasting venous blood was analysed for lipid, glucose and insulin concentrations. At the beginning and end of energy restriction, mononuclear cells were isolated from whole blood and TNF-alpha production measured by ELSIA. RESULTS: TNF-alpha production was not associated with the degree of adiposity but was higher in diabetic subjects (P<0.04). There was a reduction after energy restriction (281+/ 43 to 182+/-30 pg/ml, P<0.05) however the presence of type 2 diabetes did not influence the magnitude of this change. Plasma glucose and insulin levels decreased after weight loss in all subjects and weak correlations were found with TNF-alpha concentrations (r=0.3, P<0.05). CONCLUSIONS: We conclude that maximal production of TNF-alpha from mononuclear cells decreases with energy restriction and is weakly associated with plasma glucose and insulin concentrations in obese patients. PMID- 14757290 TI - Clustering of components of the metabolic syndrome and risk for development of type 2 diabetes in Japanese male office workers. AB - To investigate the effects of the clustering of components of the metabolic syndrome (MS) on development of diabetes, we examined 3298 Japanese male office workers aged 35-59 years who did not have type 2 diabetes (a fasting plasma glucose level of > or =7.0 mmol/l or receipt of hypoglycemic medication) or a history of cardiovascular disease. Fasting plasma glucose levels were measured at periodic annual health examinations from May 1994 through May 2001. After adjustment for potential risk factors for diabetes, the multivariate-adjusted relative risk of type 2 diabetes compared with the subjects without components of the MS was 1.58 (95% CI: 1.08-2.32), 2.48 (95% CI: 1.69-3.63), 3.10 (95% CI: 2.05 4.68), and 5.22 (95% CI: 3.49-7.83) (P-value for trend <0.001) for those with 1, 2, 3, and > or =4 components, respectively. Even after the subjects were stratified according to fasting plasma glucose level, the clustering of components of the MS was associated with an increased risk of type 2 diabetes for subjects in all three categories of low-normal fasting glucose (a fasting plasma glucose level of <5.1 mmol/l), high-normal fasting glucose (a fasting plasma glucose level of 5.0-6.0 mmol/l), and impaired fasting glucose (a fasting plasma glucose level of 6.1-6.9 mmol/l). These results indicate that clustering of components of the MS associated with diabetes precedes an increase in the risk of type 2 diabetes in Japanese men. PMID- 14757291 TI - Patients with type 2 diabetes aged 35-64 years at four primary health care centres in Stockholm County, Sweden. Prevalence and complications in relation to gender and socio-economic status. AB - This study estimates the prevalence of known diabetes, and complications of type 2 diabetes, among subjects aged 35-64 years, in relation to socio-economic factors and gender at four primary health care centres (PHCCs) in Stockholm County, Sweden. A total of 685 diabetic subjects aged 35-64 years of age were identified in primary care by diagnosis using electronic patient records. Data were supplied from medical records, and postal questionnaires. The prevalence of known diabetes among subjects aged 35-65 years differed between the PHCCs, with standardised rates between 1.7 and 3.6%, with the highest figure at the PHCC in an underprivileged area. The prevalence was higher among men (2.8% versus 1.7%). A subgroup of the patients, i.e. 389 subjects aged 35-64 years with type 2 diabetes, was studied with regard to complications. Higher education level was protective for microvascular complications (OR 0.50; CI 0.25-1.00). In addition, microvascular complications were predicted by male sex, duration, HbA(1c), and hypertension, and macrovascular complications by male sex, age, and hypertension. Prevalence of known diabetes and microvascular complications in type 2 diabetes, are associated with lower socio-economic status and male sex. PMID- 14757292 TI - Idiopathic sudden hearing loss in patients with type 2 diabetes. AB - The aim of the present study was to identify clinical and audiologic characteristics of idiopathic sudden hearing loss (ISHL) in patients with type 2 diabetes. We retrospectively investigated 148 cases of ISHL, whose age was more than 40 years, comparing clinical and audiologic valuables between diabetic and non-diabetic patients. Twenty-four patients (16.2%) had type 2 diabetes (16 male, 8 female). Prevalence of hypertension and hyperlipidemia were significantly greater in diabetic patients. Hearing in the affected ear was more impaired in diabetic than non-diabetic patients, although hearing in the unaffected ear and degree of recovery did not differ significantly. Mean BMI, duration of diabetes, HbA1c values, and ultrasonographically determined carotid intima-media thickness (IMT) and plaque scores in diabetic patients with ISHL were 24.0+/-3.7 kg/m(2), 9.8+/-7.8 years, 7.8+/-1.5%, 0.83+/-0.16 mm, and 3.8+/-2.8, respectively. Of 17 diabetic patients whose ISHL was treated with steroids, 12 required insulin for glycemic control during treatment. Compared with diabetic patients without ISHL, HbA1c value was significantly higher in diabetic patients with ISHL (7.2+/-1.2% versus 7.8+/-1.5%, P=0.0202). In conclusion, nearly 16% of our patients with ISHL had type 2 diabetes, and this subgroup was associated with more severe hearing loss. Further studies are needed to determine which subgroups of diabetic patients are most likely to develop ISHL, which patients are predisposed to more severe hearing loss, and how various factors and treatments influence outcome. PMID- 14757293 TI - Combination therapy with rosiglitazone and glibenclamide compared with upward titration of glibenclamide alone in patients with type 2 diabetes mellitus. AB - AIM: To compare the efficacy of combination therapy using rosiglitazone (8 mg per day) and glibenclamide (7.5 mg per day) with upward titration of glibenclamide as monotherapy (maximum dose=15 mg per day) in reducing HbA(1c) levels over 26 weeks in patients with type 2 diabetes mellitus (T2DM). METHODS: Three hundred and forty patients with T2DM inadequately controlled (FPG > or =7.0 and < or =15.0 mmol/l) on glibenclamide 7.5 mg per day were randomised to either additional treatment with rosiglitazone 8 mg per day or up-titration of the glibenclamide dose (maximum dose=15 mg per day). RESULTS: After 26 weeks, treatment with rosiglitazone combination reduced HbA(1c) by 0.81% (P<0.0001) and FPG by 2.4 mmol/l (P<0.0001) compared with glibenclamide monotherapy. HOMA-S and HOMA-B increased by 12 and 28%, respectively (P<0.0001 for both) with combination compared with glibenclamide monotherapy. With rosiglitazone combination and glibenclamide monotherapy, total cholesterol: HDL ratio reduced by 5 and 13%, triglycerides reduced by 6 and 2%, and FFAs reduced by 15 and 8%, respectively. Both treatments were well tolerated and had predictable safety profiles. CONCLUSION: For patients inadequately controlled on glibenclamide, addition of rosiglitazone provides significantly improved glycaemic control compared with uptitration of glibenclamide. This may be preferable to continued monotherapy with higher doses of glibenclamide. PMID- 14757294 TI - Lipoma and opthalmoplegia in mitochondrial diabetes associated with small heteroplasmy level of 3243 tRNA(Leu(UUR)) mutation. AB - We report a patient with mitochondrial diabetes mellitus associated with the A3243G mutation (MDM3243). The patient is a 77-year man with diabetes. At age 68, he noticed diplopia, due to superior rectus muscle palsy of the right eye. At age 70, he noticed lipoma on the right arm. The pathology of his muscle revealed some ragged-red fibers, and focal cytochrome c oxidase deficiency. Hence, he may have a pathogenetic mechanism in common with CPEO (chronic progressive external ophthalmoplegia) or mitochondria-related autoimmune disorder associated with mononeuropathy. He had the rate of 0.102% for heteroplasmy of 3243 mitochondrial DNA mutation in leukocytes. This case's heteroplasmy level is the smallest among the reported cases of MDM3243 in the literature. 3243 mitochondrial DNA mutation is known to induce a lack of uridine-modification in tRNA(Leu(UUR)) at the first letter of the anticodon, with which the third letter of the codon pairs, and decline of the pairing of the anticodon of tRNA with the codon of mRNA, suggesting the termination of polypeptide-elongation to generate premature proteins. Therefore, we speculate that these premature proteins may accumulate overtime, thereby affecting cells in target organs. PMID- 14757295 TI - Treatment patterns and outcomes of depressed medically ill and non-medically ill patients in community psychiatric practice. AB - The prevalence of depression among the medically ill, the recognition of depression in general medical practice, and the association between depression and medical illness have all been a focus for research in recent years. Less is known about the process and outcomes of depression care in the medically ill compared with the non-medically ill, but some studies suggest that those with concomitant physical illness have poorer outcomes. In a study of community psychiatric practice, a sample of 53 patients with no medical comorbidity (NMI) was compared with 50 patients, categorized by higher (HMI) or lower (LMI) levels of physical comorbidity, approximately 5 months after beginning treatment for a current episode of major depression. No differences were found in treatments received or in mental health outcomes between the three groups. The HMI group showed greater impairment in social and occupational functioning at baseline and significantly greater improvement in these variables at follow-up. Since medical comorbidity does not appear to adversely affect treatment decisions or outcomes in community psychiatric practice, depressed, physically ill patients should be encouraged to seek treatment, regardless of their medical condition or level of disability. PMID- 14757296 TI - Somatization disorder in young adult population. AB - Somatization is a widespread problem in health care. We estimated the occurrence of Somatization Disorder (SD) using three different case-finding methods in a general population cohort. The sample consists of 1,598 subjects born in 1966. The case-finding methods according to the DSM-III-R criteria for SD were: 1) Finnish Hospital Discharge Register (FHDR) data, 2) analysis of the patient records in public outpatient care 1982-1997, and 3) Structured Clinical Interview for DSM-III-R (SCID) for 321 selected cases. The prevalence of SD was 1.1% (N = 18), giving a female-to-male ratio of 5:1. All cases were found among the public outpatient care records. No cases appeared in the FHDR or were recognized in the psychiatric interview. The lifetime prevalence of SD was comparable with previous western population studies. Methodologically, information from outpatient records may be more sensitive in detecting SD than hospital diagnosis or even psychiatric interview. Clinically we stress the importance of recognizing these cases by liaison psychiatrists especially because SD has been recognized as being difficult to treat among somatic and primary health service providers and because some promising treatment alternatives such as cognitive-behavioral therapy and antidepressants have emerged for SD patients. PMID- 14757297 TI - Quality of life assessments in major depressive disorder: a review of the literature. AB - According to the DSM-IV classification, a diagnosis of Major Depressive Disorder (MDD) is possible only when there is evidence of significant inference with functioning. However, despite the high prevalence of MDD in the general population, it is uncommon for clinicians to assess overall functioning in a systematic way before making such diagnosis. An important correlate of functioning is quality of life, which is typically defined as "patients' own assessments of how they feel about what they have, how they are functioning, and their ability to derive pleasure from their life activities". In the present article, we review studies focusing on the relationship between depression and quality of life, particularly focusing on the impact of the treatment of depression on quality of life. Studies focusing on the quality of life in MDD are reviewed. Candidate studies published between 1970 and recently were initially identified by Pubmed and Ovid search cross-referencing the terms "quality of life," "psychosocial functioning" with "major depression" and "treatment." A number of studies report poorer quality of life in MDD patients compared to controls. Several studies also report an improvement in quality of life measures during various phases of treatment with antidepressants and/or psychotherapy. However, trials comparing the role of newer psychopharmacologic agents in the acute phase of treatment, and the role of newer psychotherapies in the continuation and maintenance phases of treatment in restoring psychosocial functioning and improving the quality of life in MDD are lacking. Exploring the impact of these modalities on psychosocial function and quality of life in MDD are necessary to help translate clinical response into restoration of psychosocial function and to thus further improve the standard of care. PMID- 14757298 TI - Psychiatric comorbidity and inpatient treatment history in bulimic subjects. AB - Bulimia nervosa (BN) is often associated with other forms of psychopathology. There is a need to clarify which specific factors of psychopathology are linked with the referral to psychiatric or psychotherapeutic inpatient treatment. This study examined which factors of psychopathology are linked with the referral of BN patients to inpatient treatment while controlling for history of suicide attempts and history of underweight. 126 females with a current diagnosis of BN purging type were assessed with the Structured Clinical Interview for DSM-IV and interviewed about their history of treatment for the BN, history of weight, and history of suicide attempts. Logistic regressions were conducted to examine whether psychiatric comorbidity, suicide attempts, and underweight were associated with inpatient treatment history. Axis I comorbidity in general, but no specific axis I disorder, was associated with inpatient history. Axes II comorbidity, especially Cluster B disorders and to a lesser degree depressive/negativistic personality disorders, was associated with a history of inpatient treatment. History of suicide attempts was also linked with inpatient experience, but history of underweight was not. The results showed that BN patients with specific types of comorbidity are more likely to be hospitalized than others. PMID- 14757299 TI - Prevalence of major depressive disorder among Chinese-Americans in primary care. AB - An epidemiological study in Los Angeles showed that Chinese Americans had lower rates of depression compared to the U.S. national estimates. This study surveys the prevalence of major depressive disorder (MDD) among Asian-Americans in the primary care setting. A two-phase epidemiological survey was performed in the primary care clinic of a community health center in Boston, MA, which provides treatment to under-served Asian-Americans. Participants were Chinese Americans in the waiting area of the primary care clinic, 18 years of age or older, who spoke any one the four commonly used Chinese dialects. The Chinese version of the Beck Depression Inventory (CBDI) was used for initial screening. All consenting patients who screened positive (CBDI >/= 16) and a fraction of those who screened negative (CBDI < 16) were interviewed by a bilingual and bicultural psychiatrist with the Structured Clinical Interview for DSM-III-R, patient version, for confirmation of the diagnosis of MDD. There were 815 in the primary care clinic that were approached, of which 503 patients (62% female, mean age 50 +/- 17 years) filled out the CBDI in the initial phase of depression screening. Extrapolating the results from the SCID-P interviews, the prevalence of MDD among Asian-Americans in the primary care setting was estimated to be 19.6% +/- 0.06. MDD is common among Asian-Americans in primary care settings. The prevalence of MDD is comparable to or higher than those found in the U.S. nonminority populations. PMID- 14757300 TI - Detection and documentation of dementia and delirium in acute geriatric wards. AB - Detection of cognitive impairment among hospitalized older individuals has shown to be insufficient. A point prevalence study in two geriatric hospitals in Helsinki, Finland, was performed among 219 acutely ill individuals over 70 years to assess the detection of dementia and delirium. Documentation of dementia and delirium in medical records, and recordings of confusional symptoms in nurses' notes were compared with the researchers' diagnosis made after a detailed assessment of cognitive status. The cognitive decline was mentioned in medical records in 70/88 (79.5%) of the cases. Cognitive testing was performed on 42/88 (47.7%) of the dementia patients, and the diagnosis of dementia was recorded in 47/88 (53.4%) of them. A specific etiological diagnosis was recorded in only 4/88 (4.5%) cases. Cognitive impairment in at least one of these four means was recorded in 80/88 (90.9%) of cases (sensitivity 0.93). Eight patients had a false positive diagnosis of dementia (specificity 0.94). Delirium was diagnosed in 77 (35.2%) patients by the researchers, but it was recorded in only 31/77 (40.3%) in medical records. In 64/77 (83.1%) cases signs of confusion were recorded in nurses' notes. Poor detection and documentation may lead to undertreatment of both disorders. PMID- 14757301 TI - Emergency department management and outcome for self-poisoning: a cohort study. AB - Self-poisoning in adults is an important public health problem across the world, but evidence to guide psychological management is lacking. In the current cohort study we wished to investigate whether aspects of routine Emergency Department management such as receiving a psycho-social assessment, or being referred for specialist follow up, affected the rate of repetition of self-poisoning. The study was carried out in four inner city hospitals in Greater Manchester, United Kingdom, over a 5-month period. We used hospital information systems and reviewed the case notes of every patient presenting to the Emergency Department to identify prospectively all adult patients presenting with deliberate self poisoning. Data regarding the Emergency Department management of each episode were collected. The Manchester and Salford self-harm database was used to determine the number of individuals who went on to repeat self-poisoning within 6 months of their index episode. During the recruitment period 658 individuals presented with self-poisoning. Traditional risk factors for repetition such as substance dependence, psychiatric contact, and previous self-poisoning were associated with a greater likelihood of receiving a psycho-social assessment or being referred for specialist follow-up. Ninety-six patients (14.6%) repeated self-poisoning within 6 months of their index episode. After adjustment for baseline demographic and clinical characteristics and hospital, receiving a psycho-social assessment was not associated with reduced repetition but being referred for specialist follow-up was [adjusted hazard ratio for repetition (95% CI): 0.49 (0.25 to 0.84), P=.01]. We found that being referred for active follow up after self-poisoning was associated with a reduced risk of repetition. The implications of this finding are discussed. Further studies using both cohort and randomized controlled study designs will help inform management strategies for patients who poison themselves. PMID- 14757302 TI - Use of restraints and pharmacotherapy in academic psychiatric emergency services. AB - Psychiatric emergency services (PES) are an increasingly important component of mental health services. To assess the type and scope of services delivered in the PES setting, the American Association for Emergency Psychiatry sponsored an Expert Consensus Panel Survey of these services in 1999. The questionnaire was mailed to medical directors of PES facilities with 91% (n = 51) responding. More than 90% of the respondents were teaching sites. Restraints were reportedly used in a mean of 8.5% of presentations for a mean of 3.3 h per episode. Restraint utilization correlated with the percentage of psychotic patients treated, but not with a wide variety of other patient and service variables. Involuntary medications were used in 16% of cases, though in oral form in 29% of those cases. A large majority (94%) endorsed mild sedation permitting further assessment as the appropriate endpoint and rejected sleep or heavy sedation as an endpoint (82%). Benzodiazepines received the strongest endorsements and 82% indicated it would be appropriate to administer a benzodiazepine alone for agitation first and initiate antipsychotic treatment subsequently if appropriate. When there is no history of prior antipsychotic exposure, 60% favored a benzodiazepine alone. However, given a history of previous antipsychotic treatment, only 8% endorsed this strategy. Most respondents (78%) preferred to use oral medication for treating behavioral emergencies, whenever possible but 70.3% reported regular use of an IM combination of a benzodiazepine and high-potency typical neuroleptic when necessary. In addition to managing emergencies, 82% of services initiated standing medications for patients being admitted to hospital settings and 70% initiated regular medication treatments for patients being released to the community. Of patients started on oral antipsychotics, 42% received an atypical antipsychotic. Reflecting medication characteristics of particular importance in emergency settings, most respondents (92%) cited selective serotonin reuptake inhibitors as the preferred type of antidepressant, and divalproex or related compounds (90%) for treatment of bipolar disorder in the PES. PMID- 14757303 TI - Social learning, affective state and passive coping in irritable bowel syndrome and inflammatory bowel disease. AB - The association between the use of passive coping strategies to deal with pain and reported levels of anxiety, depression, and parental reinforcement of illness behavior was examined in individuals with Irritable Bowel Syndrome (IBS) and Inflammatory Bowel Disease (IBD). Individuals with IBS and IBD recruited primarily from outpatient clinics completed questionnaire measures of pain-coping (the Vanderbilt Pain Management Inventory, VPMI) as well as measures of anxiety and depression, parental reinforcement of illness behavior and physical symptoms. Factor analysis of the passive coping sub-scale of the VPMI indicated that it was comprised of two components corresponding to emotional and behavioral facets of passive coping. Higher levels of behavioral passive coping were associated with higher levels of parental reinforcement of illness behavior and higher levels of depression, but only amongst individuals with IBS. In contrast emotional passive coping was associated in both groups with higher levels of anxiety and depression (but not illness-related social learning). Different factors predict the use of emotional and behavioral passive coping strategies in IBS and IBD. It is suggested that illness-related social learning occurring during childhood influences the development of habitual illness behaviors and that, because of the more benign nature of symptoms in IBS, individuals with IBS may be more likely than individuals with IBD to revert to such habitual behaviors to cope with symptoms. The degree to which the emotional component of passive coping, associated with psychological distress in both groups, can be considered in terms of 'coping strategies', rather than markers of illness-related distress, is discussed. PMID- 14757304 TI - Hospitalized patients and alcohol: who is being missed? AB - Research and clinical experience have shown that alcohol use disorders are neither sufficiently identified nor addressed in hospitalized patients. This study sought to quantify and localize these missed opportunities. The setting was an urban medical center with a Level 1 trauma designation. The only eligibility requirement was a Blood Alcohol Level (BAL) greater than 300 ng/dl upon hospital admission, a "nonsubtle" value more than three times the legal intoxication limit. Charts [58] were retrospectively reviewed for treating service (medical, trauma services, or psychiatric) and evidence of psychological signs or behavioral symptoms of withdrawal. Also assessed were the presence or absence of withdrawal monitoring, withdrawal prophylaxis orders, inpatient addictions consultation, and referral for addictions aftercare. Numerous patients with admission BALs >300 failed to be identified as needing assessment for alcohol related disorders. Patients admitted to medical or psychiatric services were significantly more likely to be diagnosed than those on trauma services (P =.02). Patients on medical or psychiatric services were also more likely to be assessed for withdrawal and referred for after-care (P <.0001) than those cared for on trauma services. The delivery of care for alcohol-related disorders was deficient, particularly for patients with traumatic injuries, even among patients severely intoxicated at admission. Failure to identify such patients represented a missed opportunity to address this vital contributor to trauma. It is suggested that both the origins of this shortfall and its resolution depend not just upon trauma providers but upon the entire medical system. PMID- 14757305 TI - The determinants of attempted suicide in a general hospital setting in Fiji Islands: a gender-specific study. AB - The objective of the study was to examine the sociodemographic and clinical factors influencing gender-specific attempted suicide. Suicide attempters, 40 males and 88 females, seen on a consultation-liaison psychiatric service over a period of 42 months (from January 1, 1999 to June 30, 2002) were compared on sociodemographic and clinical variables. The female attempters [22.99 years (SD 8.1)] were younger than their male counterparts [25.15 years (SD 9.5)] [P = 0.0002]. A higher proportion of the male attempters were engaged in outside occupations, compared with their female counterparts who were mainly full-time housewives or engaged in domestic duties [P = 0.003]. Alcohol misuse was more associated with male suicide attempts [P = 0.001]. Ninety percent of male attempters and 54.6% of the female attempters had the intention to die [P = 0.001]. Although depressive episode was the most common psychiatric disorder in male attempters, followed by neurotic and stress-related disorders, while neurotic and stress-related disorders ranked first in female attempters followed closely by depressive episode, the difference was not significant. However, significantly more male attempters required pharmacotherapy and psychological intervention than their female counterparts [P = 0.002]. More females had marital difficulties, although comparing the two groups on social difficulties did not yield a significant difference. Method used, personality disorders, previous attempt, repeat attempt, race, religion, and marital status were not significant distinguishing factors. Younger age, lack of employment outside home, marital problems, and nondeath motives were more influential in female attempted suicide, while alcohol misuse and severe psychiatric morbidity were more frequently associated with male attempted suicide. The findings support gender-specific preventive and interventional strategies. PMID- 14757306 TI - Acceptance of a trauma-focused survey: do personality and health matter? AB - Health evaluations after trauma are often performed by postal surveys, although previous studies show that some participants experience distress reactions afterwards. The aim was to explore how former burn patients react to filling in a trauma-related survey and whether the reactions are related to individual factors. The survey contained 307 questions, of which one was an open question to elicit reactions to participation. Personality was measured with the Swedish universities Scales of Personality, health with the Burn Specific Health Scale Brief, and psychological health with the Hospital Anxiety and Depression Scale and Impact of Event Scale-Revised. Participants were 78 (67%) adult burn patients, injured on average 3.9 years previously. Three groups of reactions were identified: positive/beneficial (55%), effort/time-consuming (32%), and negative/intrusive (13%). Only four participants expressed that the survey had been intrusive. Negative reactions were associated with maladaptive personality traits, poorer relationships, and more stress symptomatology, but not with burn severity or sociodemographic variables. Patients with self-inflicted injuries were evenly spread across the groups, but those with negative reactions were responsible for most of the group differences in individual factors. While a small subgroup reacted negatively, the majority accepted the trauma-focused survey and even found it beneficial. PMID- 14757307 TI - Globus hystericus: a brief review. AB - Globus hystericus, a form of conversion disorder, is characterized by an uncomfortable sensation of a mass in the esophagus or airway. Evaluation proves no mass exists. Anxiety or psychological conflict is judged to be significantly related to the onset and progression of the sensation. The sensation may lead to difficulty swallowing or breathing and may become severe or life threatening. The disorder is poorly studied and understood. The differential diagnosis is vast. Management of the disorder is similar to that suggested for other conversion disorders. This article reviews the current literature about diagnosis, etiology, treatment, and prognosis of globus hystericus. PMID- 14757308 TI - Iodoform poisoning: an unrecognized cause of consciousness disturbance. PMID- 14757309 TI - Neuropsychological performance in obsessive-compulsive disorder: a critical review. AB - There is growing evidence for neuropsychological dysfunction in obsessive compulsive disorder (OCD) related to an underlying frontal lobe and/or basal ganglia dysfunction. The following paper is a systematical review of the existing literature on cognitive impairment in OCD patients. Fifty studies were surveyed with regard to methodological aspects and cognitive impairments found in OCD patients. In addition, the impact of confounding variables such as psychotropic medication, co-morbidity or severity of symptoms on neuropsychological functioning as well as effects of treatment are discussed. OCD is often related to memory dysfunction that seems to be associated with impaired organization of information at the stage of encoding. Several other executive functions are also commonly disturbed, though results are inconsistent. The results of our study suggest that some cognitive deficits seem to be common in OCD, but future studies should focus more on possible confounding variables such as co-morbidity or psychotropic medication. PMID- 14757310 TI - Increases in lipids and immune cells in response to exercise and mental stress in patients with suspected coronary artery disease: effects of adjustment for shifts in plasma volume. AB - This study examined the role of shifts in plasma volume on lipid and immune reactions to stress. Lipid, immune, rheological, and cardiovascular reactions to exercise and mental stress in 51 patients with suspected coronary artery disease were determined. Blood pressure and heart rate were measured during and blood samples taken at the end of each rest and task. Lipids (total cholesterol, triglycerides, HDL, LDL) and immune cells (lymphocytes, monocytes, granulocytes) increased with exercise, whereas cholesterol, LDL, and lymphocytes increased with mental stress. Plasma volume decreased by 1 and 5% following mental and exercise stress, respectively. The task-induced increases in lipids were no longer statistically significant following adjustment for changes in plasma volume, whereas the increases in immune cell numbers survived such correction. This study provides evidence that, in coronary artery disease patients, exercise and mental stress-induced increases in lipids but not immune cells can be largely accounted for by shifts in plasma volume. PMID- 14757311 TI - Performance and sleepiness during a 24 h wake in constant conditions are affected by diet. AB - This study investigated the effects of high-carbohydrate (HC) and high-fat (HF) diet on cognitive performance, and subjective and objective sleepiness. Seven male participants were kept awake for 24 h in a metabolic ward. Meals were given every 4h and cognitive performance and sleepiness ratings were assessed hourly. The Karolinska Drowsiness Test (KDT, EEG derived) was performed twice after meal. Performance in simple reaction time showed a significant interaction of diet and the post-prandial period, a slower reaction time was observed for the HC-diet 3.5 h after meal intake. Diet did not affect EEG measures but a general post-prandial increase of objective sleepiness was observed 3.5h after meal servings. The HC diet was significantly associated with an increase of subjective sleepiness. The study demonstrated that the HC-diet caused larger oscillation in performance and increased sleepiness as compared to HF-diet throughout day and night. PMID- 14757312 TI - Electrophysiological evidence of an early effect of sentence context in reading. AB - Recognition Potential is an electrophysiological response of the brain that is sensitive to semantic aspects of stimuli. According to its peak values (about 250ms), Recognition Potential appears as a good candidate to reflect lexical selection processes. Consequently, Recognition Potential might be sensitive to contextual information during reading a sentence. In present study, the standard procedures to improve the visibility of Recognition Potential (Rapid Stream Stimulation paradigm) were used in a task in which sentence context was crucial. A parieto-occipital Recognition Potential was observed to peak about 264ms after stimulus onset, followed by a centro-parietal N400 peaking at about 450ms. Recognition Potential was affected by contextual information though, contrary to N400, presenting larger amplitude to contextually congruous words. These results support the assumption that Recognition Potential may reflect lexical selection processes, representing also evidence of context effects on ERP around 250ms after stimulus onset during sentence reading. PMID- 14757313 TI - Artemisinin derivatives: toxic for laboratory animals, safe for humans? AB - A discrepancy seems to prevail with regard to the toxicity and safety of the artemisinin family of antimalarials. While these compounds have been found to be virtually void of any serious side effects in humans, their neurotoxicity in animal models has raised concerns about their use. In this paper, we present selected examples of both pre-clinical and clinical studies dealing with adverse effects of artemisinin drugs. We suggest that the prolonged presence of artemisinins upon slow release from oil-based intramuscular formulations is the main cause of the observed toxicity in laboratory animals. In contrast, oral intake of these compounds, which is by far the most common formulation used for treatment of malaria patients, results in rapid clearance of these drugs and is thus unlikely to cause any toxicity in human subjects. Another plausible factor may be the relatively high doses of artemisinin compounds used in animal studies. In conclusion, the observation of the toxicity of artemisinin compounds in animals, but not in humans, is most likely due to different pharmacokinetic profiles after different routes of administrations. PMID- 14757314 TI - Inhibitory effect of tellimagrandin I on chemically induced differentiation of human leukemia K562 cells. AB - Tellimagrandin I is a hydrolysable tannin compound widely present in plants. In this study, the effect of tellimagrandin I on chemically induced erythroid and megakaryocytic differentiation was investigated using K562 cells as differentiation model. It was found that tellimagrandin I not only inhibited the hemoglobin synthesis in butyric acid (BA)- and hemin-induced K562 cells with IC50 of 3 and 40microM, respectively, but also inhibited other erythroid differentiation marker including acetylcholinesterase (AChE) and glycophorin A (GPA) in BA-induced K562 cells. Tellimagrandin I also inhibited 12-O tetradecanoylphorbol-13-acetate (TPA)-induced expression of CD61 protein, a megakaryocytic marker. RT-PCR analysis showed that tellimagrandin I decreased the expression of erythroid genes (gamma-globin and porphobilinogen deaminase (PBGD)) and related transcription factors (GATA-1 and NF-E2) in BA-induced K562 cells, whereas tellimagrandin I induced the overexpresison of GATA-2 transcription factor that played negative regulation on erythroid differentiation. These results indicated that tellimagrandin I had inhibitory effects on erythroid and megakaryocytic differentiation, which suggested that tannins like tellimagrandin I might influence the anti-tumor efficiency of some drugs and the hematopoiesis processes. PMID- 14757315 TI - Hydropic degeneration of the anterior pituitary gland (adenohypophysis) in uremic rats. AB - We observed hydropic degeneration of the anterior pituitary in rats made uremic by nephrotoxic chemicals, especially when the uremic rats were given a pure carbohydrate diet beforehand. The hydropic degeneration caused loss of nuclear and cytoplasmic content of many or most anterior pituitary cells. It was readily visible in paraffin sections by light microscopy. It was exaggerated when water was injected after the nephrotoxin and it was greatly reduced if saline was injected after the nephrotoxin. Low serum sodium levels in affected rats and the response to saline injection suggested that the mechanism for development of hydropic degeneration of the anterior pituitary gland involved hyponatremia. Depletion of total body sodium probably accounts for the enhancement of hydropic degeneration by the pure carbohydrate diet. Morphologic lesions of the anterior pituitary related to hyponatremia and uremia have not been described previously. PMID- 14757316 TI - Reactions of acrylamide with glutathione and serum albumin. AB - Rate constants of 0.0054 and 0.021 M(-1)s(-1) for the reactions of acrylamide with human serum albumin (HSA) and glutathione (GSH), respectively, were determined under physiological conditions by following the loss of their thiol groups in the presence of excess acrylamide. Based on these in vitro values, reactions with these thiols appear to account for most of acrylamide's elimination from the body. Combined with data from other studies, these results should be useful for assessing the health risk of dietary acrylamide. PMID- 14757317 TI - Tissue-specific effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on the activity of 5'-deiodinases I and II in rats. AB - Thyroid hormones play a complex role in the toxicity of polychlorinated dibenzo-p dioxins and furans and related compounds. We investigated the toxicological significance of 5'-deiodinases I and II (5'-DI and 5'-DII) in the altered thyroid hormone status of TCDD-treated rats. Time courses and dose responses were determined for serum thyroxine (T4) and triiodothyronine (T3) levels, for 5'-DI activity in thyroid gland, liver and kidney, and for 5'-DII activity in brown adipose tissue (BAT). TCDD-treatment resulted in prompt and dose-dependent decrease in circulating T4 followed by a decrease in liver 5'-DI activity 1-2 days later and an apparent increase in BAT 5'-DII activity. Changes in liver 5' DI and BAT 5'-DII activity were secondary to decreased T4 levels. Thyroid and kidney 5'-DI activities as well as circulating T3 levels were not affected. The results suggest that altered 5'-DI or 5'-DII activities do not significantly influence the circulating levels of T4 or T3 in TCDD-treated rats. PMID- 14757318 TI - Cytotoxicity of chromium and manganese to lung epithelial cells in vitro. AB - Chromium, nickel and manganese are the predominant metals in welding fumes and are associated through epidemiological studies with an increased risk for developing occupational asthma due to welding activities. Here, we show that chromium(VI) and manganese, but not nickel, are cytotoxic to normal human lung epithelial cells in vitro (SAEC and BEAS-2B), at concentration ranges of 0.2-200 microM. The toxic effect was associated with increased levels of intracellular phosphoprotein and subsequent release of inflammatory cytokines IL-6 and IL-8, while no release of TNF-alpha was observed. Changes in intracellular phosphoprotein levels occurred at concentrations below the cytotoxic effect. IL-6 and IL-8 production increased up to 4.4-fold relative to controls. IL-6 and IL-8 are released from lung epithelium to recruit cells of the immune system to sites of tissue damage. Therefore, the observed effects of chromium(VI) and manganese in lung epithelial cells demonstrate a mechanism through which the toxicity of these metals to epithelial cells can result in recruitment of cells of the immune system. PMID- 14757319 TI - Acute-phase protein expression in DMSO-intoxicated rats. AB - The ability of dimethyl sulfoxide (DMSO) to induce the acute-phase (AP) response was examined. Injection of DMSO to laboratory rats caused a rapid doubling of the plasma corticosterone concentration 2 h after treatment. The elevated corticosterone concentration promoted the synthesis of mRNAs for several AP reactants. At 24 h after DMSO administration the relative serum concentration of cysteine-proteinase inhibitor (CPI) increased about 710%, alpha1-acid glycoprotein (AGP) 630%, alpha1-macroglobulin (MG) 510%, gamma fibrinogen (Fb) 420%, haptoglobin (Hp) 280%, whereas the relative concentration of albumin, a "negative" AP reactant, decreased to 93%. The extent and kinetics of the corticosterone increase and the general increase of AP reactant mRNAs and protein serum concentrations after DMSO administration corresponded to the overall changes observed during the turpentine-induced AP response. On the basis of these findings it was concluded that DMSO was capable of promoting the AP response in rats. PMID- 14757320 TI - Linoleic acid epoxide promotes the maintenance of mitochondrial function and active Na+ transport following hypoxia. AB - Low concentrations of arachidonic acid monoepoxides protect against ischemia/reperfusion injury. This study examined whether low concentrations of the linoleic acid monoepoxide, cis-12,13-epoxy-9-octadecenoic acid (12,13-EOA), protect renal cells against decreases in mitochondrial and transport functions induced by hypoxia/reoxygenation. Primary cultures of rabbit renal proximal tubular cells (RPTC) were pretreated with diluent or 1, 5, or 10 microM 12,13-EOA for 1 h and exposed to 2 h hypoxia/0.5 h reoxygenation in the absence of 12,13 EOA. Basal respiration, oligomycin-sensitive oxygen consumption (QO2), and ATP content decreased 31, 35 and 65%, respectively, following hypoxia/reoxygenation. Hypoxia/reoxygenation also increased mitochondrial membrane potential (DeltaPsi(m)). Pretreatment with 12,13-EOA prevented decreases in basal and oligomycin-sensitive QO2s and increases in DeltaPsi(m). Despite the protection against decreases in mitochondrial function, 12,13-EOA pretreatment did not prevent the initial decrease in intracellular ATP content following hypoxia. However, pretreatment did accelerate the recovery of intracellular ATP levels during reoxygenation. Pretreatment with 12,13-EOA also prevented hypoxia-induced decreases in active Na+ transport. Ouabain-sensitive QO2 (a marker of active Na+ transport) decreased 38% following hypoxia/reoxygenation but was maintained in RPTC pretreated with 1, 5 or 10 microM 12,13-EOA prior to hypoxia. Pretreatment of RPTC with the hydrolyzed product of 12,13-EOA, 12,13-dihydroxyoctadecenoic acid, did not have any protective effects against mitochondrial dysfunction and decreases in active Na+ transport. Thus, this is the first report demonstrating that preconditioning of RPTC with low concentrations of 12,13-EOA, but not its hydrolyzed product, maintains mitochondrial respiration, accelerates restoration of ATP levels, and prevents decreases in active Na+ transport following hypoxia/reoxygenation. PMID- 14757321 TI - Benzylmercapturic acid is superior to hippuric acid and o-cresol as a urinary marker of occupational exposure to toluene. AB - The present study was initiated to examine whether urinary benzylmercapturic acid (or N-acetyl-S-benzyl cysteine, BMA), a mercapturate metabolite of toluene, increases in relation to the intensity of toluene exposure, and whether this metabolite is a better marker of occupational exposure to toluene than two traditional markers, hippuric acid and o-cresol. Accordingly, end-of-shift urine samples were collected from 122 printers and 30 office clerks (all men) in the second half of a working week. Solvent (toluene) exposure of the day (8 h) was monitored by means of diffusive sampling. Quantitative relation with toluene showed that BMA had a greater correlation coefficient with toluene (r = 0.7) than hippuric acid (r = 0.6) or o-cresol (r = 0.6). The levels in the urine of the non exposed control subjects were below the detection limit of 0.2 microg/l for BMA, whereas it was at substantial levels for hippuric acid and o-cresol (239 mg/l and 32 microg/l as a geometric mean, respectively). Thus, BMA, hippuric acid and o cresol could separate the exposed from the non-exposed when toluene was at < 1, 50 and 3 ppm, respectively. Overall, therefore, it appeared reasonable to conclude that BMA is superior to hippuric acid and o-cresol as a marker of occupational exposure to toluene. PMID- 14757322 TI - Influence of altered apoptosis in human lymphoblastoid cell lines on micronucleus frequency. AB - Defects in apoptosis play a decisive role in both tumorigenesis and drug resistance in tumor treatment. The purpose of this study was to investigate the balance between formation of genomic damage and induction of apoptosis upon genotoxic stress. For this, we influenced the apoptotic response and measured the amount of genomic damage expressed as micronucleus formation after treatment with the topoisomerase II inhibitor etoposide. Apoptosis was reduced by the addition of pifithrin (PFT) alpha and enhanced by transient transfection with bcl-2 antisense-oligonucleotide in Bcl-2-overexpressing cells. We used three human lymphoblastoid cell lines with different p53 status (TK6, wild-type p53; WTK1, mutated p53; NH32, p53 double knockout). Under conditions of reduced apoptosis, micronucleus formation was also reduced. When apoptosis was increased, micronucleus formation remained unchanged or was also increased. Overall, we did not find an expected inverse correlation between induction of apoptosis and genomic damage. PMID- 14757324 TI - Decreased cerebrospinal fluid neuropeptide Y (NPY) in patients with treatment refractory unipolar major depression: preliminary evidence for association with preproNPY gene polymorphism. AB - Extensive animal studies suggest neuropeptide Y (NPY) to be involved in coping with a wide range of stressors, and that impaired central NPY signalling could be involved in the pathophysiology of anxiety and depression. Human studies of central NPY levels in depression have, however, been inconclusive. Here, we examined levels of NPY-like immunoreactivity (NPY-LI) in the cerebrospinal fluid (CSF) of medication-free subjects with treatment refractory unipolar depression. Patients were admitted to a research inpatient unit, examined under standardized conditions, and compared with a sample of volunteers in whom psychiatric morbidity was excluded. A robust suppression of NPY levels in patient CSF was found, while other putative CSF markers (monoamine metabolites, somatostatin) did not differ between the groups. We then explored whether this finding might be related to a recently described T1128C coding polymorphism which results in a Leu7-> Pro7 substitution of the signal peptide, and a previously not described T 399C polymorphism in the promoter region of the preproNPY gene. Preliminary evidence was found for an association of both markers with a diagnosis of depression, indicating the possibility of an underlying haplotype influencing the vulnerability for developing depressive illness. Our present findings are in line with an extensive animal literature, and further support the notion that impaired NPY function could contribute to depressive illness. PMID- 14757325 TI - Combined DEX/CRH test among Japanese patients with major depression. AB - There is compelling evidence for an important role of the hypothalamus-pituitary adrenal (HPA) axis abnormalities in the pathophysiology of major depressive disorder. Growing evidence has suggested that the combined dexamethasone (DEX)/CRH test is much more sensitive than the conventional DEX suppression test in order to detect HPA axis abnormalities. However, little data is currently available on DEX/CRH results for Asian populations, which prompted us to examine the sensitivity of the DEX/CRH test among Japanese subjects with major depression. The DEX/CRH test was administered in 20 inpatients with major depressive episode and 30 healthy controls. Significantly increased cortisol responses were observed for the patients, compared to the controls. There was a substantial difference in the distribution of non-suppressor, intermediate suppressor, and suppressor, which were defined in terms of cortisol response, was observed between the patients and controls (10, 60, and 30% in the patients vs. 0, 27, and 73% in the controls, P<0.01). Responses of ACTH showed a trend in the same direction. Within the depressed patients, individuals with a history of attempted suicide, in particular, tended to have enhanced responses to the DEX/CRH test, compared to those without such a history. Our results confirmed that the DEX/CRH test is a sensitive test to detect HPA axis abnormalities among Japanese patients with major depression. In addition, a possible relationship between suicidal acts and enhanced HPA axis abnormalities was suggested. PMID- 14757327 TI - Verbal memory performance of patients with a first depressive episode and patients with unipolar and bipolar recurrent depression. AB - Depression is usually associated with episodic memory impairment. The main clinical features of depression associated with that memory impairment are not clearly defined. The main goal of that study was to assess the role of the diagnostic subtypes and the number of depressive episodes on the memory performance of acute unipolar (UP) and bipolar (BP) depressed patients.Twenty three patients with a first major depressive episode (FE), 28 patients meeting DSM-IV criteria for UP recurrent depression (UR) and 18 BP patients with recurrent depression were compared with 88 healthy subjects on a verbal episodic memory task. Patients suffering from a first depressive episode did not show verbal memory impairment as compared to normal controls. Unlike FE patients, UR and BP patients exhibited verbal memory deficits with impaired free recall and normal cued recall and recognition. The memory deficits of the UR and BP patients was present in the first free recall trial. Depressed patients improved their memory performance across the three trials of the task at the same rate than normal controls. Our results suggest that the number of depressive episodes has a negative influence on verbal memory performance of acute depressed patients. The effects of the repetition of the depressive episodes are not modulated by the subtypes of depression and may reflect sensitization to the cognitive impact of depression associated with increasing prefrontal dysfunction. PMID- 14757326 TI - Treatment with the CRH1-receptor-antagonist R121919 improves sleep-EEG in patients with depression. AB - Well documented changes of sleep electroencephalogram (EEG) in patients with depression include rapid eye movement (REM) sleep disinhibition, decreases of slow-wave-sleep (SWS) and increase in wakefulness. Twenty-seven inpatients with major depression were admitted subsequently to a clinical trial with the CRH(1) receptor-antagonist R121919 administered in two different dose escalation panels. A random subgroup of 10 patients underwent three sleep-EEG recordings (baseline before treatment, at the end of the first week and at the end of the fourth week of active treatment). SWS time increased significantly compared with baseline after 1 week and after 4 weeks. The number of awakenings and REM density showed a trend toward a decrease during the same time period. Separate evaluation of these changes for both panels showed no significant effect at lower doses, whereas in the higher doses after R121919 REM density decreased and SWS increased significantly between baseline and week 4. Furthermore positive associations between HAMD scores and SWS at the end of active treatment were found. Although these data might indicate that R121919 has a normalizing influence on the sleep EEG, the design of the study does not allow to differentiate genuine drug effects from those of clinical improvement and habituation to the clinical setting. PMID- 14757328 TI - Differential impairment on measures of attention in patients with paranoid and nonparanoid schizophrenia. AB - The purpose of the present study is to investigate whether patients with different subtypes of schizophrenia are differentially impaired on measures of attention. Forty-eight patients with schizophrenia (19 paranoid and 29 nonparanoid) and 48 healthy controls (matched on chronological age, sex, and years of education) were administered five measures of attention including the Stroop Color-Word Test (SCWT; Stroop, 1935), the Digit Vigilance Test (DVT; Lewis, 1992), the Symbol Digit Modalities Test (SDMT; Smith, 1982), the Backward Digit Span Test (BDST; Wechsler, 1987), and the Color Trails Test (CTT; D'Elia et al., 1996) to assess selective attention, sustained attention, switching attention, and attentional control processing by the latter two tests respectively. Results from the present study showed that patients with schizophrenia performed poorer on the SCWT, the DVT, and the SDMT, relative to their healthy counterparts. Furthermore, patients with different subtypes of schizophrenia also had different degrees of attentional impairment. While patients with paranoid schizophrenia performed worse on the SCWT, those with nonparanoid schizophrenia performed worse on the SDMT. Nevertheless, these findings may suggest that patients with paranoid and nonparanoid schizophrenia may have different profiles with respect to their performances on measures of attention. PMID- 14757329 TI - Superior temporal gyrus and P300 in schizophrenia: a combined ERP/structural magnetic resonance imaging investigation. AB - Decrement of the auditory P300 component of the event-related potentials (ERP) is a robust finding in schizophrenic patients and seems to be most pronounced in the left temporal region. Structural MRI studies support the hypothesis that regional structural brain differences in this patient group include reduced volume in temporal lobe structures. The aim of the presented study was to investigate the possible gray matter volume reductions in the left posterior superior temporal gyrus (STG) and the P300 reduction and left 5 degrees ) (non-physiological) abduction angular perturbations of the human knee. This reflex action was shown to originate from periarticular tissue afferents. Furthermore, it was demonstrated that specific muscle activation patterns exist in knee muscles with preferential activation in medial muscles in response to the lateral perturbations. This study examines the hypothesis that in response to the mechanical stimulus, the sensory information mediated by these afferents results in activation patterns that provide the largest resisting moment by the knee muscles. It is further hypothesized that this near maximum resistance cannot be achieved by the selective activation of medial muscles alone. To examine this, the previously reported mechanically induced reflex EMG activation patterns, a stochastic 3D musculoskeletal patello-femoral joint model, and new data from selective electrical stimulation experiments were used. Using the model, the knee adduction-abduction moment in response to an applied abduction load at the knee joint for every possible random set of quadriceps activity was computed. These adduction moments were then compared to the adduction moment computed by the model when the mechanically induced muscle activation patterns were used. The data presented here illustrated that selective activation of a medial muscle alone would result in an abduction moment, regardless of the knee flexion angle. Furthermore, the findings of this study revealed that the recorded combinations of muscle activity provide a near maximum capability of the quadriceps muscles to resist externally applied abducting stimuli. It was concluded that stabilization in the abduction direction could only be achieved by a control strategy that involves activation of both medial and lateral muscles at the knee. It was also concluded that this control strategy was near optimal when mediated by joint afferents. PMID- 14757340 TI - The use of a muscle graft to repair a segmentary nerve defect. An experimental study using the sciatic nerve of rats as model. AB - The use of a devitalized skeletal muscle graft and conventional nerve graft to repair a 5mm long segmentary sciatic nerve lesion was studied in rats by means of functional, morphometric and spinal cord motor neuron cell response evaluation. Thirty-four rats were used and divided into four groups according to the procedure: (1) sham operation; (2) conventional nerve grafting; (3) muscle grafting; (4) unrepaired lesion. The sciatic functional index (SFI) was evaluated every fortnight up to the 105th postoperative day by measuring three parameters in the rats' footprint. The animals of Groups 2 and 3 presented initial complete functional loss, followed by slow but steady recovery, with final similar SFIs. The histologic and morphometric studies showed an increased small diameter/thin myelin sheath nerve fiber density distally to the lesion site for both types of graft. An increased population of motor neurons was observed in the anterior horn of the lumbar spinal cord segment with both types of grafts, but not in the control groups. The SFI, histologic and morphometric data did not differ significantly between the two types of graft, thus indicating a similar behavior. The authors conclude that a 5mm long skeletal muscle graft works as well as a conventional nerve graft. PMID- 14757341 TI - A low power multichannel analog front end for portable neural signal recordings. AB - We present the design and testing of a 16-channel analog amplifier for processing neural signals. Each channel has the following features: (1) variable gain (70-94 dB), (2) four high pass Bessel filter poles (f(-3 dB)=445 Hz), (3) five low pass Bessel filter poles (f(-3 dB)=6.6 kHz), and (4) differential amplification with a user selectable reference channel to reject common mode background biological noise. Processed signals are time division multiplexed and sampled by an on-board 12-bit analog to digital converter at up to 62.5k samples/s per channel. The board is powered by two low dropout voltage regulators which may be supplied by a single battery. The board measures 8.1 cm x 9.9 cm, weighs 50 g, and consumes up to 130 mW. Its low input-referred noise (1.0 microV(RMS)) makes it possible to process low amplitude neural signals; the board was successfully tested in vivo to process cortically derived extracellular action potentials in primates. Signals processed by this board were compared to those generated by a commercially available system and were found to be nearly identical. Background noise generated by mastication was substantially attenuated by the selectable reference circuit. The described circuit is light weight and low power and is used as a component of a wearable multichannel neural telemetry system. PMID- 14757342 TI - A multichannel telemetry system for single unit neural recordings. AB - We present the design, testing, and evaluation of a 16 channel wearable telemetry system to facilitate multichannel single unit recordings from freely moving test subjects. Our design is comprised of (1) a 16-channel analog front end board to condition and sample signals derived from implanted neural electrodes, (2) a digital board for processing and buffering the digitized waveforms, and (3) an index-card sized 486 PC equipped with an IEEE 802.11b wireless ethernet card. Digitized data (up to 12 bits of resolution at 31.25k samples/s per channel) is transferred to the PC and sent to a nearby host computer on a wireless local area network. Up to 12 of the 16 channels were transmitted simultaneously for sustained periods at a range of 9 m. The device measures 5.1 cm x 8.1 cm x 12.4 cm, weighs 235 g, and is powered from rechargeable lithium ion batteries with a lifespan of 45 min at maximum transmission power. The device was successfully used to record signals from awake, chronically implanted macaque and owl monkeys. PMID- 14757343 TI - Development of rapid staining protocols for laser-capture microdissection of brain vessels from human and rat coupled to gene expression analyses. AB - Laser-capture microdissection (LCM) is a technique that enables selective extraction of desired cells from heterogeneous tissues compatible with subsequent molecular analyses. The specific visualization of desired cell types prior to LCM is essential for achieving selective capture. We have developed rapid and selective staining protocols for LCM extraction of microvessels from human and rat brain. Vessels in human and rat brain sections were visualized by a 2 min exposure to fluorescein-labeled lectins Ulex Europeaus Agglutinin I (UEA I) and Ricinus Communis Agglutinin I (RCA I), respectively. Immunohistochemical staining for the endothelial-specific marker, Factor VIII-related antigen (FVIII-rAg), co localized with that for either UEA I or RCA I, confirming the selective staining of vascular structures with these lectins. Both brain vessels and perivascular parenchyma were captured using LCM, followed by RNA isolation. RT-PCR analyses demonstrated the enrichment of LCM-captured vessels and parenchyma in FVIII-rAg and GFAP mRNA, respectively. LCM-captured human vessels also expressed the tight junction-specific gene, zonula occludens 1 (ZO-1). LCM extraction of vessels from brain sections can be used to perform molecular fingerprinting of neurovascular unit in various brain pathologies. PMID- 14757344 TI - Large-scale microarray gene expression analysis in discrete electrophysiologically identified neuronal clusters. AB - The normal processes of learning and memory as well as the pathological progress of various neurological diseases may result in changes in gene expression in small, local populations of neurons in any given brain area, leading to the occurrence of specific patterns of electrical activity without easily detectable changes in the morphology of this brain area. One way of identifying these changes might be the comparison of gene expression of areas which generate and areas which do not generate specific patterns of electrical activity. A method for microbiopsy of limited (0.5-1.0 mm3) tissue samples from electrophysiologically identified areas of neurons generating epileptiform activity in the rat brain is described. Here we demonstrate that total RNA isolated from individual microbiopsy samples might be successfully used for microarray based gene expression analysis of any discretely localized neuronal group which can be identified electrophysiologically, including neurons in cortical columns, cell assemblies or other functional units. PMID- 14757345 TI - A multi-channel telemetry system for brain microstimulation in freely roaming animals. AB - A system is described that enables an experimenter to remotely deliver electrical pulse train stimuli to multiple different locations in the brains of freely moving rats. The system consists of two separate components: a transmitter base station that is controlled by a PC operator, and a receiver-microprocessor integrated pack worn on the back of the animals and which connects to suitably implanted brain locations. The backpack is small and light so that small animal subjects can easily carry it. Under remote command from the PC the backpack can be configured to provide biphasic pulse trains of arbitrarily specified parameters. A feature of the system is that it generates precise brain stimulation behavioral effects using the direct constant-voltage TTL output of the backpack microprocessor. The system performs with high fidelity even in complex environments over a distance of about 300 m. Rat self-stimulation tests showed that this system produced the same behavioral responses as a conventional constant-current stimulator. This system enables a variety of multi-channel brain stimulation experiments in freely moving animals. We have employed it to develop a new animal behavior model ("virtual" conditioning) for the neurophysiological study of spatial learning, in which a rat can be accurately guided to navigate various terrains. PMID- 14757346 TI - Chronic lumbar catheterization of the spinal subarachnoid space in mice. AB - A simple method for chronic intrathecal (i.t.) catheterization of the lumbar subarachnoid space in mice is described. The procedure does not require major surgery and does not produce neurological deficits. The intrathecal catheter stayed in place and was functional for at least 10 days. Morphological studies revealed no histological damage in the spinal cord after catheter implantation. The effects of acute and chronic intrathecal morphine were studied and compared between the current method and that of acute lumbar puncture. Morphine produced similar antinociceptive effect when administered acutely. All mice with catheters responded reliably to daily morphine injection up to 5 days whereas it was difficult to inject drugs repeatedly with lumbar puncture. It is concluded that this novel method of chronic lumbar catheterization in mice has advantages over the existing lumbar puncture technique for intrathecal delivery of drugs upon repeated administration. This method may be particularly useful in studies of genetically modified mice where the number of mice available is often limited. PMID- 14757347 TI - Design and validation of a computer-based sleep-scoring algorithm. AB - A computer-based sleep scoring algorithm was devised for the real time scoring of sleep-wake state in Wistar rats. Electroencephalogram (EEG) amplitude (microV(rms)) was measured in the following frequency bands: delta (delta; 1.5-6 Hz), theta (Theta; 6-10 Hz), alpha (alpha; 10.5-15 Hz), beta (beta; 22-30 Hz), and gamma (gamma; 35-45 Hz). Electromyographic (EMG) signals (microV(rms)) were recorded from the levator auris longus (neck) muscle, as this yielded a significantly higher algorithm accuracy than the spinodeltoid (shoulder) or temporalis (head) muscle EMGs (ANOVA; P=0.009). Data were obtained using either tethers (n=10) or telemetry (n=4). We developed a simple three-step algorithm that categorizes behavioural state as wake, non-rapid eye movement (NREM) sleep, rapid eye movement (REM) sleep, based on thresholds set during a manually-scored 90-min preliminary recording. Behavioural state was assigned in 5-s epochs. EMG amplitude and ratios of EEG frequency band amplitudes were measured, and compared with empirical thresholds in each animal.STEP 1: EMG amplitude greater than threshold? Yes: "active" wake, no: sleep or "quiet" wake. STEP 2: EEG amplitude ratio (delta x alpha)/(beta x gamma) greater than threshold? Yes: NREM, no: REM or "quiet" wake. STEP 3: EEG amplitude ratio Theta(2)/(delta x alpha) greater than threshold? Yes: REM, no: "quiet" wake. The algorithm was validated with one, two and three steps. The overall accuracy in discriminating wake and sleep (NREM and REM combined) using step one alone was found to be 90.1%. Overall accuracy using the first two steps was found to be 87.5% in scoring wake, NREM and REM sleep. When all three steps were used, overall accuracy in scoring wake, NREM and REM sleep was determined to be 87.9%. All accuracies were derived from comparisons with unequivocally-scored epochs from four 90-min recordings as defined by an experienced human rater. The algorithms were as reliable as the agreement between three human scorers (88%). PMID- 14757348 TI - Sindbis vector SINrep(nsP2S726): a tool for rapid heterologous expression with attenuated cytotoxicity in neurons. AB - Sindbis virus-based vectors have been successfully used for transient heterologous protein expression in neurons. Their main limitation arises from infection-associated cytotoxicity, attributed largely to a progressive shut down of host cell protein synthesis. Here we evaluated a modified Sindbis vector, based on a viral strain containing a point mutation in the second nonstructural protein, nsP2 P726S, described to delay inhibition of protein synthesis in BHK cells [Virology 228 (1997) 74], for heterologous expression in neurons in vitro and in vivo. First, we constructed an optimized helper vector, termed DH BB(tRNA/TE12), for production of SINrep(nsP2S(726)) viral particles with low levels of helper RNA co-packaging and high neurospecificity of infection. Second, we determined that hippocampal primary neurons infected with SINrep(nsP2S(726)) virus expressing EGFP showed a delayed onset of viral induced cytotoxicity and higher levels of EGFP expression in comparison to cells infected with wild type SINrep5 EGFP-expressing virus. However, a strong decrease in protein synthesis still occurred by day 3 postinfection. The SINrep(nsP2S(726)) vector is thus well suited for rapid high level expression within this time window. As an experimental example, we demonstrate the applicability of this system for high resolution two-photon imaging of dendritic spines in vivo. PMID- 14757349 TI - Transport of a synaptotagmin-YFP fusion protein in sympathetic neurons during early neurite outgrowth in vitro after transfection in vivo. AB - Developing neurons are engaged in neurite outgrowth as well as the synthesis and transport of proteins involved in synaptic transmission. Very little is known about when transport is established in these rudimentary neurites. We used a novel technique to visualize protein transport during the early hours of neurite outgrowth in culture. Recombinant adenoviruses were used to express a synaptotagmin-YFP fusion protein in the superior cervical ganglia of neonatal rats in vivo and protein transport was examined in neuronal cultures established from the superior cervical ganglions (SCGs). We find that, as early as 4h in culture, synaptotagmin-YFP was present in the cytoplasm, lamellipodia, filopodia and growth cones. Protein expression appeared punctate in neurites at 8h in vitro and is consistent with a vesicular localization. These results indicate that the machinery to transport synapse-specific proteins is functional in rudimentary neurites at this time and indicates that this technique can be used to study early neuronal development. PMID- 14757350 TI - Quantification of neurons in the myenteric plexus: an evaluation of putative pan neuronal markers. AB - Accurate estimates of the total number of neurons located in the wall of the gut are essential for studies of the enteric nervous system (ENS). Though several stains and antibodies are used routinely as pan-neuronal markers, controversies of relative sensitivity and completeness have been difficult to resolve, at least in part because comparisons often must be made across experiments and laboratories. Therefore, we evaluated the efficacy of four putative pan-neuronal markers for the ENS, under comparable conditions. Neurons in the myenteric plexus of wholemounts taken from the small intestines of Fischer 344 rats were stained using Cuprolinic Blue, anti-HuC/D, anti-protein gene product 9.5, or FluoroGold injections followed by permanent labeling with an antibody to the FluoroGold molecule. All four markers had useful features, but both protein gene product 9.5 and FluoroGold were found to be problematic for obtaining reliable counts. As a result, only neurons labeled with either Cuprolinic Blue or anti-HuC/D were compared quantitatively. Based on counts from permanently labeled tissue, Cuprolinic Blue and HuC/D were similarly effective in labeling all neurons. Because the two protocols have different strengths and weaknesses, Cuprolinic Blue and HuC/D provide a complementary set of labels to study the total neuronal population of the ENS. PMID- 14757351 TI - An ultra small array of electrodes for stimulating multiple inputs into a single neuron. AB - We have developed an ultra small, translucent array of electrodes for use in the parasaggital cerebellar slice preparation. This positionable array is capable of stimulating multiple independent bundles of parallel fibers (PFs), which synapse onto a single Purkinje neuron. On a silicon substrate, a low-stress silicon nitride film was used both as a structural layer and as electrical insulation. Evaporated gold pads and interconnects were sandwiched between two such layers. A bulk anisotropic silicon etch released the individual arrays. The electrodes are supported within a 2-microm-thick cantilever of translucent silicon nitride. In one design, eight 4-microm-wide square electrodes are arranged on 8-microm centers. Another design, half the scale of the first, was also tested. The array was mounted on a micromanipulator and can be visualized by an upright microscope. It can then be positioned in the dendritic arbor of a Purkinje neuron while not disturbing a recording pipette at the soma. Paired-pulse facilitation experiments have confirmed that the electrodes are capable of stimulating non-overlapping bundles of PFs. This device will be useful for exploring spatiotemporal synaptic integration in single neurons. Potential applications in experiments on cerebellar LTD are also discussed. PMID- 14757352 TI - Continuous wavelet transform in the evaluation of stretch reflex responses from surface EMG. AB - OBJECTIVE: This is the first reported use of the continuous wavelet transform (CWT) of the surface EMG (sEMG) to extract the reflex response to muscle stretch. We used a modulus-based method to estimate instantaneous amplitude-envelopes from ridges of the CWT (referred in this work as sEMG intensity) to extract the dynamic reflex response from sEMG. We tested the method on tendon reflexes where excellent temporal resolution is required to identify the different latency components, and on the tonic stretch reflex (tonic SR) response to an ongoing perturbation that characteristically has a low signal to noise ratio. METHODS: Eight subjects without neurological impairment were subjected to a series of archilles tendon taps and a 2 min continuous perturbation of the ankle using a pseudo-sinusoidal stretch profile containing frequencies from 0.1 to 8.0 Hz. The tendon reflexes were assessed in the soleus muscle at 10% of MVC and the tonic SR in tibialis anterior while the muscle was relaxed, at 5 and 10% of maximal voluntary contraction. Root mean square (RMS) and wavelet ridge extraction was applied to the sEMG signal to extract sEMG amplitudes (RMS) and intensities for all reflexes. To obtain the tonic SR, these estimates and those from the sEMG-RMS were subsequently cross-correlated with the perturbation record to yield 2 sets of estimates of reflex gain and coherence for comparison. RESULTS: The sEMG intensities were highly correlated with the torques resulting from a ramped voluntary contraction. Following tendon taps, the method resolved the M1, M2, M3 response components at accurate latencies and with more complete reconstruction of the components than RMS-derived estimates. The wavelet ridge estimates extracted the tonic SR from resting and contracting muscles with significantly higher coherence than RMS estimates. Reflex gain, when estimated from sEMG intensity or sEMG-RMS, demonstrated similar relationships to the perturbation frequency and background contraction level. When the sEMG intensity reflex gain estimates from different subjects were pooled, they showed significantly lower variance about the mean than gain estimates derived from the rectified sEMG. CONCLUSIONS: Wavelet-ridge extraction provides a valid approach to reflex evaluation from sEMG that does not depend on the absolute amplitude of the potentials measured at the EMG electrodes. This may have substantial advantages in more directly comparing responses between subjects on an absolute frequency scale without the need for normalisation against maximal contraction levels. PMID- 14757353 TI - Methods for chronic neural recording in the telencephalon of freely behaving fish. AB - We have adapted for use in fish several of the procedures employed for recording single neuron activity in freely behaving rodents. Developing a method for single unit chronic recording in freely behaving fish was motivated by a need for a comparison across taxa of telencephalic neural activity evoked during spatial navigation by animals of their environments. However, the procedures outlined here can be modified easily for underwater recording from most aquatic species and from other brain areas. Under anesthesia, bundles of stereotrodes or tetrodes were implanted into the dorsolateral region of the goldfish or cichlid telencephalon. An infrared light emitting diode (LED) was also fixed to the fish's head at the time of surgery. After recovery from anesthesia, fish were allowed to swim freely within a large aquarium. Single unit activity was analyzed and correlated with stimulus conditions, behavior, and the location and movement of the LED recorded by a camera tracking system. The value of this technique is demonstrated by providing the first evidence in fish for navigation-related neural firing, including "place cells" that display location-specific discharge. PMID- 14757354 TI - Conditioned spikes: a simple and fast method to represent rates and temporal patterns in multielectrode recordings. AB - Increasing evidence suggests that the brain utilizes distributed codes that can only be analyzed by simultaneously recording the activity of multiple neurons. This paper introduces a new methodology for studying neural ensemble recordings. The method uses a novel representation to provide complementary information about the stimuli which are contained in the temporal pattern of the spike sequence. By using this procedure, a high correlation of synchronized events with stimuli times is apparent. To quantify the results and to compare the performance of this method against the most traditional raster plot, we have used Fano factor and cross-correlation analysis. Our results suggest that several consecutive spikes from different neurons within an extended time window may encode behaviorally relevant information. We propose that this new representation, in addition to the other approaches currently used (standard raster plots, multivariate statistical methods, neuronal networks, information theory, etc.), can be a useful procedure to describe population spike dynamics. PMID- 14757355 TI - Simultaneous surface electromyography (SEMG) and 31P-MR spectroscopy measurements of the lumbar back muscle during isometric exercise. AB - The aim of this study was to demonstrate the feasibility of simultaneous surface electromyography (SEMG) and 31P-MR spectroscopy (31P-MRS) measurements on the back muscle of volunteers during the performance of an isometric exercise. Six volunteers (three male, three female) performed a modified Biering-Sorensen test inside a 1.5 T MR scanner while simultaneously recording SEMG signals. A surface coil was used for 31P-MRS with a CSI sequence. Spectra were collected with a voxel resolution of 40 mm x 40 mm x 100 mm and a temporal resolution of 30 s during periods of rest, sustained muscle contraction and recovery. The duration of muscle contraction was 150 s. SEMG analysis yielded a decrease of the mean SEMG frequency of approximately 20%. The SEMG amplitudes were constant or increased up to approximately 150% during exercise. 31P-MRS showed a maximum decrease of the phosphocreatine (PCr) amplitude down to approximately 32% of its initial value. Simultaneously, a doubling of the inorganic phosphate (Pi) signal was observed. The present study demonstrates that simultaneous SEMG and 31P-MRS measurements of the back muscle are feasible during isometric exercises. PMID- 14757356 TI - Confocal microscopic imaging of fast UV-laser photolysis of caged compounds. AB - Using a pulsed UV laser in a confocal scanning microscope, we present a relatively cheap, accurate and efficient method for fast UV laser flash photolysis of caged molecules in two-dimensional cultured neurons. The laser light is introduced through the imaging optics, can be localized by a parallel red laser and can photolyse a sphere of less than 1 microm2, and evoke local fluorescence changes in the imaged neurons. Caged glutamate and caged fluorescein are used to illustrate a disparity between spines and their parent dendrites at a sub-micron resolution. PMID- 14757357 TI - The dynamics of spatial behavior: how can robust smoothing techniques help? AB - A variety of setups and paradigms are used in the neurosciences for automatically tracking the location of an animal in an experiment and for extracting features of interest out of it. Many of these features, however, are critically sensitive to the unavoidable noise and artifacts of tracking. Here, we examine the relevant properties of several smoothing methods and suggest a combination of methods for retrieving locations and velocities and recognizing arrests from time series of coordinates of an animal's center of gravity. We accomplish these by using robust nonparametric methods, such as Running Median (RM) and locally weighted regression methods. The smoothed data may, subsequently, be segmented to obtain discrete behavioral units with proven ethological relevance. New parameters such as the length, duration, maximal speed, and acceleration of these units provide a wealth of measures for, e.g., mouse behavioral phenotyping, studies on spatial orientation in vertebrates and invertebrates, and studies on rodent hippocampal function. This methodology may have implications for many tests of spatial behavior. PMID- 14757358 TI - Activity patterns as a correlate for sleep-wake behaviour in mice. AB - Sleep-wake behaviour in mice is known to interact with various behavioural dimensions. Therefore, it is necessary to control for such dimensions when evaluating sleep in mice. The characterisation of sleep in rodents usually is based on EEG signals. Since this method demands the invasive implantation of electrodes, it cannot be integrated into general behavioural phenotyping procedures. Thus, non- or minimum-invasive methods are needed for the analysis of sleep-wake behaviour. Although physiological parameters, like for instance general locomotor activity, allow for the assessment of sleep-wake behaviour in mice, existing methods lack reliability especially in measuring stationary and three-dimensional activities. In this study, a small magnet was implanted subcutaneously near the neck muscles of mice and each movement of the magnet was registered via a sensor plate. For validation of the described method, the effects of sleep deprivation were evaluated by both the magnet and the EEG in parallel. Our results show that the data obtained via the subcutaneously implanted magnet represent a reliable and sensitive measurement of quantitative aspects of sleep-wake behaviour: spatial variation as well as stationary activities could be dissociated from sleep. Qualitative sleep characteristics were not detected. In summary, this minimum invasive method allows for the detection of quantitative alterations in sleep-wake behaviour in mice, thus, offering a useful, rapid pre-screen in animal sleep research. PMID- 14757359 TI - Nanoprobe NMR spectroscopy and in vivo microdialysis: new analytical methods to study brain neurochemistry. AB - Nuclear magnetic resonance (NMR) spectroscopy was used to study the chemical composition of cerebrospinal fluid (CSF) microdialysate from the rat brain. In vivo microdialysis techniques were used in several brain regions including the frontal cortex, amygdala, striatum, nucleus accumbens and third ventricle and dialysate samples (20microl) were subsequently analyzed by one and two dimensional 1H NMR experiments using a Varian nanoprobe. Neurochemical resonances were assigned on the basis of published chemical shifts [Lindon et al., Ann. Rep. NMR Spectrosc. 38 (1999) 1-88], correlation experiments and addition of standard compounds. Glucose, lactate, formate, pyruvate, creatinine, gamma hydroxybutyrate, acetate, glutamate, glycine, tyrosine, isoleucine, leucine, alanine and choline were some of the neurochemicals unambiguously assigned. Additional studies in the frontal cortex showed that amino acids such as glutamate, alanine and isoleucine were sensitive to local tetrodotoxin (TTX) infusion. The NMR spectra were also subjected to multivariate statistical methods to compare the different brain regions examined. To our knowledge, the present experiments are the first to describe the combination of nanoprobe NMR technology with in vivo microdialysis for the analysis of brain neurochemistry in freely moving rats. PMID- 14757360 TI - Electrochemical monitoring of transport by a vesicular monoamine transporter expressed in Xenopus oocytes. AB - Xenopus laevis oocytes were injected with synthetic mRNA coding for a rat VMAT2 mutant (rVMAT2-I483A/L484A) shown previously to be retained on the plasma membrane as a result of a presumed reduction of endocytosis. Binding of the specific VMAT inhibitor [3H]dihydrotetrabenazine indicated that expression did occur at a level of approximately 3 fmol per oocyte. To determine if rVMAT2 I483A/L484A expressed in oocytes was capable of substrate transport, oocytes were placed in buffer at pH 6.0, dopamine substrate was injected into the cell, and egress of substrate was monitored by fast scan cyclic voltammetry using a carbon fiber microelectrode. Under these conditions, transport by oocytes injected with RNA coding for rVMAT2-I483A/L484A ranged from approximately 0.5 to more than 2.5 pmol/min. Water-injected and uninjected control oocytes did not exhibit appreciable transport activity. Transport by rVMAT2-I483A/L484A-injected oocytes was reduced to control levels by tetrabenazine, a known inhibitor of VMAT transport activity. Comparison of subtracted voltammograms obtained from transport assays with those for calibration experiments confirmed that the transported species was dopamine. These results suggest that expression of VMATs in oocytes may provide a useful model system for mechanistic and regulatory studies that would not be feasible using traditional methods. PMID- 14757361 TI - A multichannel electronic monitor of acoustic behaviors, and software to parse individual channels. AB - We designed an electronic device to monitor calling behavior in male crickets. The device and its associated software can record to disk the activity of as many as 16 individuals simultaneously. The data recorded contain detailed information about the temporal structure of each individual's calls. The temporal resolution achieved with an ordinary PC is good enough to detect and record the occurrence of every pulse of sound from each cricket. The resulting data files are efficient and compact, and so the system is appropriate for experiments lasting many days. PMID- 14757362 TI - Does AID aid AIDS? AB - During AIDS, the acquisition of mutations in the HIV-1 gp120 envelope glycoprotein leads to the switch from primary R5 (CCR5-using) to highly cytopathic X4 (CXCR4-using) HIV-1 variants. Based on the already known sequence homology between IgV genes and the gp120-coding region of the env gene, as well as on somatic hypermutation of multiple proto-oncogenes, the somatic hypermutation hypothesis for the mechanism of R5-X4 HIV-1 switching is proposed as follows. This switch takes place in the germinal center (GC) B cells due to the aberrant somatic hypermutation of the gp120-coding part of the HIV-1 env gene. Activation-induced cytidine deaminase (AID) is required for this process. Activation through IL4R and CD40 is required both for infection of GC B cells with HIV-1 and for induction of AID expression in the same cells. B cell infection with R5 HIV-1 variants is the limiting stage in the process of the viral phenotypic switch during the asymptomatic period of AIDS. Overall up regulation of CXCR4 coreceptor on the GC B cells and the CD4(+) T cells surrounding the GC provides the predominant replication and acquisition of the newly formed X4 HIV-1 variants. PMID- 14757363 TI - Physiological T cell activation starts and propagates in lipid rafts. AB - Lipid rafts are plasma membrane compartments enriched in key signaling molecules. We have previously shown that in T lymphocytes anti-CD3 stimulation is insensitive to cholesterol extraction by methyl-beta-cyclodextrin (MbetaCD), suggesting that anti-CD3 induced signal transduction is independent of raft integrity. Here we show that, in contrast to T cell stimulation by anti-CD3 antibodies, T cell activation by a physiological ligand is mediated by signaling events taking place in lipid raft. Indeed, cholesterol depletion by MbetaCD resulted in reduced T cell activation in response to Epstein Barr Virus (EBV) transformed B cells pulsed with a bacterial superantigen. Moreover, T cell stimulation by pulsed EBV-B cells, but not by anti-CD3 antibodies, induced recruitment of active Lck in detergent-resistant membranes, where the signal transduction is organized and amplified. PMID- 14757364 TI - Downregulation of class II transactivator (CIITA) expression by synthetic cannabinoid CP55,940. AB - Cannabinoid receptors are known to be expressed in microglia; however, their involvement in specific aspects of microglial immune function has not been demonstrated. Many effects of cannabinoids are mediated by two G-protein coupled receptors, designated CB1 and CB2. We have shown that the CB1 receptor is expressed in microglia that also express MHC class II antigen (J. Neuroimmunol. 82 (1998) 13-21). In our present study, we have analyzed the effect of cannabinoid agonist CP55,940 on MHC class II expression on the surface of IFN gamma induced microglial cells by flow cytometry. CP55,940 blocked the class II MHC expression induced by IFN-gamma. It has been shown that the regulation of class II MHC genes occurs primarily at the transcriptional level, and a non-DNA binding protein, class II transactivator (CIITA), has been shown to be the master activator for class II transcription. We find that mRNA levels of CIITA are increased in IFN-gamma induced EOC 20 microglial cells and that this increase is almost entirely eliminated by the cannabinoid agonist CP55,940. These data suggests that cannabinoids affect MHC class II expression through actions on CIITA at the transcriptional level. PMID- 14757365 TI - Lysophosphatidylcholine is a regulator of tyrosine kinase activity and intracellular Ca(2+) level in Jurkat T cell line. AB - Lysophospholipids, particularly lysophosphatidylcholine (lyso-PC), have been implicated in modulating T cell functions at the sites of inflammation and atherosclerosis. Although the chemotactic and immunomodulatory effects are well documented, the exact signaling pathway of lyso-PC action is poorly defined. In this work, we studied the earliest biochemical events in T cells triggered by lyso-PC. A marked and immediate tyrosine phosphorylation was induced in the leukemic T cell line, Jurkat. Phosphorylation of cellular substrates included src family kinase, p56(lck) and syk family kinase, ZAP70. The lyso-PC induced tyrosine phosphorylation was largely dependent on the presence of functional p56(lck). Tyrosine phosphorylation was followed by the elevation of intracellular Ca(2+) concentration. The magnitude of the mobilization of the intracellular Ca(2+) was similar in the absence of the p56(lck) activity in JCaM1.6 cells as in Jurkat cells, however, it was slightly but reproducibly delayed compared to that in the wild type cells. Inhibition of the Ser/Thr kinases and tyrosine kinases with staurosporine and genistein, respectively, decreased the rise in the intracellular Ca(2+) content. Moreover, pertussis toxin completely blocked the Ca(2+) signal supporting the role of the G-protein coupled LPC receptor in this event. PMID- 14757366 TI - Dissociation of CD4(+) cell counts from viral load and association with immune complexes in HIV(+) hemophilia patients. AB - OBJECTIVE: We have postulated that the host autoimmune response regulates and mediates CD4 depletion during HIV infection by opsonization of circulating CD4(+) lymphocytes carrying autoreactive immune complexes (IC) consisting of complement fixing IgM and IgG, and during advanced stages of HIV disease of IgM/ IgG/gp120 complexes. In this retrospective study, we investigated whether HIV causes CD4 depletion by direct cytotoxicity or indirectly by induction of a host autoimmune response. PATIENTS AND METHODS: In 1996, 12 HIV(+) hemophilia patients were converted to highly active antiretroviral therapy (HAART), while 10 other patients were maintained on conventional antiretroviral treatment and another 11 patients refused to be treated with antiretroviral drugs. The host immune response of these 33 HIV(+) patients was studied during the periods of minimum viral replication (Interval 1), subsequent rise in viral replication with strong replication dynamic (Interval 2), and maximum viral replication (Interval 3). The patients were categorized into three groups according to viral load (VL). Group A: patients with low level VL (n=10) showed a modest increase from <80 to <4 log 10 HIV-1 RNA copies per milliliter plasma during the observation period; Group B: patients with medium level VL (n=12) showed a stronger increase from <80 to >4 log 10 copies per milliliter plasma; and Group C: patients with high level VL (n=11) consistently had a median of >4 log 10 copies per milliliter plasma, during Intervals 1-3, with the exception of one patient who during Interval 2 had 4800 copies per milliliter. Blood lymphocyte subpopulations, proportions of CD4(+) blood lymphocytes coated with IgM, IgG, C3d and/or gp120, in vitro responses to mitogens and pooled allogeneic stimulator cells, as well as numbers of HIV-1 RNA copies per milliliter plasma were measured. RESULTS: Sequential analysis of VL, IC load on CD4(+) blood lymphocytes and CD4 counts showed that an increasing VL was not associated with CD4 depletion, when the proportion of IC coated circulating CD4(+) blood lymphocytes remained stable. When, CD4 counts and IC load were analyzed during corresponding intervals of retroviral replication in the three patient groups, a higher VL was associated with lower CD4 counts only when the IC load (IgG or gp120/IgG) on CD4(+) lymphocytes was higher as well. CONCLUSION: These data suggest that HIV regulates and mediates CD4 depletion in part by the induction of autoreactive ICs against CD4(+) lymphocytes, especially complement-fixing autoreactive IgG and gp120/IgG complexes. PMID- 14757367 TI - Serum amyloid A-induced mRNA expression and release of tumor necrosis factor alpha (TNF-alpha) in human neutrophils. AB - Recently, we described the effect of the acute phase protein serum amyloid A (SAA) on the mRNA expression and release of IL-8 in neutrophils [Mediators Inflamm. 12 (3) (2003) 173]. Here, we expand this earlier study, focusing on tumor necrosis factor-alpha (TNF-alpha) m-RNA expression and protein release. Our findings indicate that SAA stimulates the rapid expression and release of TNF alpha from cultured human blood neutrophils. The release of TNF-alpha from SAA stimulated neutrophils is strongly suppressed by the addition of the antioxidants N-acetyl-L-cysteine, alpha-mercaptoethanol, glutathione, the antiinflammatory dexamethasone and the compounds wortmannin (a PI3K inhibitor), PD98059 (a MEK-1 inhibitor) and SB203580 (a p38 inhibitor). Monocytes also responded to SAA by releasing TNF-alpha. These data are congruent with the increasing evidence of the role of SAA in modulating inflammatory and immune responses, possibly contributing to the pool of cytokines produced in acute inflammation and in chronic diseases. PMID- 14757369 TI - Lymphocyte subsets in healthy adult Kuwaiti Arabs with relative benign ethnic neutropenia. AB - Relative and absolute neutropenia is frequently seen in the healthy adult Kuwaiti Arab population. Fluorescent monoclonal antibody labelling followed by flow cytometry was used to determine the lymphocyte subsets in 48 normal healthy individuals in the Kuwaiti adult population (24 males and 24 females, age 17-59 years) with relative or absolute neutropenia, and this was compared to age matched controls (64 males and 63 females). The mean haemoglobin levels were 13.6+/-1.5 and 13.7+/-1.5 g/dl in the neutropenic and control groups, respectively. White blood cell counts, absolute neutrophil and lymphocyte counts in neutropenic individuals with the corresponding reference range, taken from the control subjects (in parenthesis) were: WBC, 6.7+/-1.6 x 10(9)/l (4-10.4 x 10(9)/l), neutrophils, 2.7+/-0.8 x 10(9)/l (1.87-6.63 x 10(9)/l), lymphocytes, 3.3+/-0.9 x 10(9)/l (1.4-3.62 x 10(9)/l). Absolute values of lymphocytes, CD2+, CD3+, CD19+, CD4+, CD8+, HLADR+ and CD45RA+ cells were significantly higher in the neutropenic group. The range of values with the corresponding reference ranges, in parenthesis, were: CD2+, 1.61-4.30 x 10(9)/l (0.95-2.99 x 10(9)/l), CD3+, 1.37-4.16 x 10(9)/l (0.83-2.71 x 10(9)/l), CD19+, 0.16-1.09 x 10(9)/l (0.05 0.61 x 10(9)/l), CD4+, 0.70-2.89 x 10(9)/l (0.45-1.65 x 10(9)/l), CD8+, 0.57-1.80 x 10(9)/l (0.29-1.17 x 10(9)/l), HLADR+ 0.27-1.74 x 10(9)/l (0.02-0.62 x 10(9)/l), CD45RA, 0.90-4.63 x 10(9)/l (0.34-2.05 x 10(9)/l), respectively. The levels of natural killer cells, CD56+ cells were significantly lower compared to controls while the values of memory T lymphocytes, CD45RO+ were comparable to controls. These results indicate that difference in the leukocyte subpopulations may also be indicative of differences in the lymphocyte subpopulations and that reference ranges for these cell types in healthy neutropenic and non-neutropenic individuals should be established. PMID- 14757368 TI - Expression of CD13/aminopeptidase N and CD10/neutral endopeptidase on cultured human keratinocytes. AB - Keratinocytes actively participate in immune response and inflammation by secreting cytokines and chemokines. Membrane-bound peptidases serve as negative loop in controlling concentration of peptide signalling molecules. Recently, they have also been proposed as additional mechanism of cell-to-cell interaction and as signalling molecules. In this study, we examined expression of two membrane bound peptidases: aminopeptidase N (APN; EC 3.4.11.2; CD13) and neutral endopeptidase (NEP; EC 3.4.24.11; CD10) on nonstimulated cultured human keratinocytes obtained from healthy skin. Membrane expression of CD13 and CD10 was analysed by FACS and fluorescent microscope. Functional properties of CD13 and CD10 were examined by testing their enzymatic activity towards selective substrates. The data were compared to those obtained on cultured nonstimulated human skin fibroblasts expressing both CD13/APN and CD10/NEP. Approximately one third (i.e. 31.7+/-2.8%; n=3) of cultured keratinocyte express CD13 as compared to fibroblasts which are 100% CD13(+) (n=3). Density of CD13 on keratinocytes is several times lower than on fibroblasts. Membrane CD13 expression on keratinocytes was associated with significant enzyme activity, which on the basis of substrate (L-Ala-betaNA) and inhibitor (bestatin, actinonin) selectivity could be ascribed to aminopeptidase N. Kinetic parameter V(max) revealed lower APN activity expressed on keratinocytes than on fibroblasts (V(max)=1.49+/-0.08 microM/60 min/5 x 10(4) cells for keratinocytes, n=3 versus V(max)=4.09+/-0.76 microM/60 min/5 x 10(4) cells for fibroblasts, n=3). Likewise, K(m) value of APN on keratinocytes was lower as compared to fibroblasts (K(m)=0.307+/-0.090 mM for keratinocytes, n=3 versus K(m)=0.766+/-0.065 mM for fibroblasts, n=3). CD13 demonstrated on cultured keratinocytes, is at least partly due to its constitutive expression since it was also found on freshly prepared epidermal skin cells. Inhibitors of APN, actinonin, bestatin and substance-P, as well as the APN blocking antibody WM-15, decreased keratinocytes growth. In contrast to membrane CD13 associated with APN enzyme activity, neither membrane CD10, nor its enzyme (NEP) activity could be found on the same keratinocyte samples. In conclusion, functional CD13, associated with APN activity, was found on about one third of cultured, non-stimulated keratinocytes, whereas no CD10/NEP was found on the same keratinocyte samples. Role of APN in regulation of keratinocyte growth is suggested, as its inhibition resulted in decreased keratinocyte growth. PMID- 14757370 TI - Comparison of adult and fetal cytokine secretion during mixed lymphocyte reactions. AB - Umbilical cord blood transplants are associated with a lower incidence of graft versus-host disease (GVHD) than adult marrow or peripheral blood stem cell transplants, and this could be related to a difference in cytokine production between fetal and adult mononuclear cells after allogeneic stimulation. Mixed lymphocyte reactions (MLRs) involving adult cells were associated with greater interferon-gamma (IFNgamma) secretion than MLRs between cord blood cells, although IL-2 secretion was similar. Experiments in which T cells were separated from accessory cells then recombined in artificial combinations indicated that differences in T cells were primarily responsible for the greater [IFNgamma]:[IL 2] ratios generally found after MLRs involving adult cells compared to fetal cells, but accessory cells also influenced this ratio. The cellular basis for the observed difference was not established, but mononuclear cell preparations from cord blood contained significantly higher proportions of CD16(+)56(-) NK-type cells and a CD19(+)1c(+) B cell subset, as well as more CD45 RA-expressing nai;ve T cells. PMID- 14757371 TI - Induction of regulatory dendritic cells by dexamethasone and 1alpha,25 Dihydroxyvitamin D(3). AB - Dendritic cells (DC) modulated to induce T cell hyporesponsiveness have promising potential in immunotherapy of autoimmune disorders and for the prevention of allograft rejection. While studying the effect of immunosuppressive agents on the maturation of DC we found that 1alpha,25-Dihydroxyvitamin D(3) the active form of Vitamin D(3) (D(3)) in combination with dexamethasone (Dex) has a synergistic effect on LPS-induced maturation of DC. Monocyte-derived DCs cultured with D(3) and Dex during LPS-induced maturation have a low stimulatory effect on allogeneic T cells comparable with that of immature DCs. But in contrast to immature DCs, D3/Dex exposed DCs secrete IL-10 and show upregulated transcription of mRNA encoding the Ig-like inhibitory receptor ILT4. D3/Dex exposed DCs also inhibit alloreactivity and slightly enhance the degree of apoptosis in mature DCs. Thus, D(3)/Dex is an effective immunosuppressive drug combination for the induction of DCs capable of inducing T cell hyporesponsiveness. PMID- 14757372 TI - CD1c(+) immature myeloid dendritic cells are predominant in cord blood of healthy neonates. AB - It has been suggested lately that some types of antigen presenting cells-myeloid dendritic (DC-1) cells can differentiate the immune response towards Th1 type immunity, whereas lymphoid cells (DC-2) can stimulate Th2 type immunity. It has been observed that neonates are deficient in Th1 response. The purpose of our study was to estimate the proportions of immature myeloid (CD1c(+)) and lymphoid (BDCA-2(+), BDCA-4(+)) dendritic cells and the CD1c(+):BDCA-2(+) cell ratio in cord blood of healthy neonates in comparison with dendritic cells of healthy adults. Thirty healthy neonates born from normal pregnancies and 30 healthy adults were included in the study. The dendritic cells were isolated from cord and peripheral blood, stained with anti-CD1c, anti-BDCA-2, anti-BDCA-4, anti CD123 and anti-CD19 monoclonal antibodies and estimated using flow cytometry. The percentage of CD1c(+) dendritic cells in cord blood of healthy newborns did not differ significantly when compared to those in peripheral blood of healthy adults. The percentages of cord blood BDCA-2(+) and BDCA-4(+) dendritic cells of neonates were significantly lower when compared to lymphoid dendritic cells in peripheral blood of adults. The CD1c(+):BDCA-2(+) ratio was significantly higher in cord blood of neonates in comparison with CD1c(+):BDCA-2(+) ratio in adult's blood. Myeloid and lymphoid dendritic cells may be involved in the immune regulation during fetal development. Immature myeloid dendritic cells are predominant in cord blood of healthy neonates. Immature lymphoid dendritic cells are not the major population of dendritic cells in cord blood. PMID- 14757373 TI - Modulation of peripheral blood T-lymphocytes in kidney transplant recipients treated with low dose OKT3 therapy. AB - The immunosuppressive effect of OKT3 depends upon both T cell depletion and antigenic modulation of CD3 complex. To establish the effect of low doses of OKT3 on peripheral T lymphocytes, we analyzed 47 kidney transplant recipients receiving OKT3 for the first time. OKT3 was used as rescue therapy in 39 patients and as part of induction protocols in 8. The mean age of patients was 39+/-10 years, 30 were females and 9 were re-transplants. Half of them (51.1%) received kidney from cadaver donors. Among those receiving OKT3 as rescue therapy, 82% recovered graft function, including patients with severe BANFF-graded rejections. After the first dose of OKT3, it a pronounced T cell depletion was observed followed by an increase in CD4 and CD8 expression in CD3 negative T cells, supporting the idea that T cell modulation was present. In conclusion, low dose OKT3 was effective in treating severe allograft rejection by inducing a sustained TCR/CD3 down modulation without long-lasting T cell depletion. PMID- 14757374 TI - Determining the threshold effect of ozone on daily mortality: an analysis of ozone and mortality in Seoul, Korea, 1995-1999. AB - Many studies have shown a positive association between ambient ozone levels and mortality. Typically, these findings are based on models that assume a linear relationship between log mortality and ozone level. In this study, we adapted generalized additive models in which ozone effects are presumed to occur in three different ways: as a simple linear term, as a cubic natural spline term, and as a combination of two linear terms (a threshold model). We applied these models to daily time-series data for Seoul, Korea for the years 1995-1999 and found that the threshold model always fits best among the three. A 2.6% (95% CI: 1.7-3.5) increase of estimated relative risk (RR) in the total mortality associated with a 21.5 ppb increase of daily 1-h maximum ozone lagged by 1 day was observed by linear Poisson's regression. However, a 3.4% (95% CI: 2.3-4.4) increase in the estimated RR was observed using the threshold model. Adjustments for other ambient pollutants caused little changes to these results; 2.4-2.5% in the linear models and 3.2-3.4% in the threshold models. In addition, the largest difference in the estimated RRs of the linear and threshold models was observed in the summer: 1.9% (95% CI: 0.5-3.3) by the linear model and 3.8% (95% CI: 2.0-5.7) by the threshold model. These findings indicate that the conventional time-series Poisson regression model, which dose not take threshold into consideration, could underestimate the true risk of the ozone effect on daily mortality. PMID- 14757375 TI - Estimating the global burden of disease of mild mental retardation and cardiovascular diseases from environmental lead exposure. AB - The disease burden from exposure to lead resulting in mild mental retardation (due to IQ point decreases) and cardiovascular outcomes (due to increases in blood pressure) was estimated at a global level. Blood lead levels were compiled from the literature for 14 geographical regions defined by the World Health Organization according to location and adult and child mortality rates. Adjustments were applied to these levels, where appropriate, to account for recent changes relating to the implementation of lead-reduction programs and the lower levels seen in rural populations. It is estimated that mild mental retardation and cardiovascular outcomes resulting from exposure to lead amount to almost 1% of the global burden of disease, with the highest burden in developing regions. This estimate can be used to assess the magnitude of the benefits that could be accrued by increasing the global coverage of lead-reduction programs. PMID- 14757376 TI - Pesticide exposure and risk of breast cancer: a nested case-control study of residentially stable women living on Long Island. AB - We conducted a nested case-control study of women in the New York State cohort who lived on Long Island. We estimated the historical environmental exposure to pesticides in 105 women diagnosed with breast cancer between 1980 and 1992 and 210 age and race-matched controls in this cohort of long-term residentially stable women who completed a short mailed questionnaire in 1980. Prior agricultural land use was assessed from aerial photographs taken in 1947 and 1950. Pesticides detected in drinking water were estimated from measures taken between 1977 and 1992. Geographic information system software was used to calculate the distance between residences and hazardous waste sites (HWS) containing pesticides. We found an increased breast cancer risk for women residing within 1 mile of HWS containing organochlorine pesticides (odds ratio [OR]=2.8; 95% confidence interval [CI], 1.1-7.1), after adjusting for other risk factors. A significant interaction was shown for women residing on land that was previously used for agriculture and women who were either nulliparous or had an older age at first birth (> or =26 yr old), compared to women who did not live on previously agricultural land and with a younger age (<26 yr old) at first birth (OR=6.4; 95%CI, 2.2-18.2). Study power was limited and confidence intervals were wide. Our findings suggest that exposure to pesticides in the environment needs to be more comprehensively investigated in relation to breast cancer risk. PMID- 14757377 TI - Weak electromagnetic fields (50 Hz) elicit a stress response in human cells. AB - The aim of this study was to demonstrate the expression of heat shock (HS) genes in human cells in response to extremely low-frequency electromagnetic fields (ELF EMF) alone and in combination with thermal stress. After exposing human myeloid leukemia (HL-60) cells to the stressor(s) for 30 min we quantified the expression of the HS genes HSP27, HSP60, HSP70 (A, B, and C), HSC70, HSP75, HSP78, and HSP90 (alpha and beta) by RT-PCR. The results clearly show that HS genes, in particular the three HSP70 genes (A, B, and C), are induced by ELF-EMF, a reaction that is enhanced by simultaneous HS (43 degrees C for 30 min). The results show similarities and some significant differences to previous experiments in which transgenic nematodes were used to monitor the induction of the HSP70 gene under similar stress conditions. We also studied the effect of different flux densities on gene expression in the range of 10-140 microT. Even the lowest dose tested (10 microT) resulted in a significant induction of the genes HSP70A, HSP70B, and HSP70C. The reaction to ELF-EMF shows a maximum at a flux density of 60-80 microT. The unusual dose-response relation reveals an interesting difference to other stressors that elicit the HS response. PMID- 14757378 TI - Apnea and bradycardia due to anaphylaxis to tobacco glycoprotein in the infant rabbit. AB - Prenatal and postnatal exposure to cigarette smoke is associated with an increased incidence of the sudden infant death syndrome, although the cause(s) for this is unknown. Tobacco glycoprotein (TGP), a group of proteins purified from cured tobacco leaves and present in cigarette smoke, have been shown to cause anaphylaxis in excised hearts and lungs of adult rabbits that were neonatally sensitized to TGP and later rechallenged. We sought to determine whether anaphylaxis occurred in live infant rabbits who were neonatally sensitized to TGP. At the age of 1 day, 12 animals were sensitized to TGP (0.1mg in 0.25 cc alum) via intraperitoneal injection (i.p.i.) followed by a booster ipi at the age of 30 days (TGP-S). Seven animals received i.p.i. of antigen-free alum only (controls). All animals underwent an intravenous TGP challenge at age 42+/-2 days. Heart rate (HR) and respiratory rate (RR) were recorded for 2 min prior to and 5 min after the challenge. Baseline HR (approximately 260) and RR (approximately 120) were similar in all animals. Seven TGP-S animals developed apnea (1.9-4.7s) within 60s of the challenge while none of the controls did. The TGP-S also became bradycardic (the lowest HR over 50 consecutive beats), with the HR decreasing from 260 to 220 vs the controls, whose HR remained constant (approximately 250). We conclude that some rabbits neonatally sensitized to TGP develop apnea and bradycardia upon further intravenous TGP challenge. These studies suggest that cigarette smoke exposure may be associated with a higher rate of SIDS via an anaphylactic mechanism. PMID- 14757379 TI - Fish consumption and other environmental exposures and their associations with serum PCB concentrations among Mohawk women at Akwesasne. AB - A study was conducted with the objective of assessing how dietary, occupational, and residential exposures to polychlorinated biphenyls (PCBs) contribute to body burden among pregnant Mohawk women residing near three hazardous waste sites. From 1992 to 1995, 111 pregnant women were interviewed about fish consumption and other environmental risk factors and donated a 20-mL venous blood sample for serum PCB analysis. To supplement previous fish sampling, samples of residential soil, ambient air, wild duck, and local meats and vegetables were also collected and analyzed for PCBs. The results indicated a significant decline in local fish consumption from an annual mean of 31.3 meals more than 1 year prior to pregnancy to an annualized mean of 11.7 meals during pregnancy. This change was reportedly a result of the advisories issued against consumption of local fish by pregnant and nursing women of childbearing age. The geometric mean concentration of total PCBs in the serum was 1.2 ppb, a level that is similar to that in other studies of women with no unusual exposures to PCBs. However, multiple regression analysis revealed that serum levels of total PCBs and three individual congeners were associated with local fish consumption. The PCB levels in soil, air, and local foodstuffs other than fish generally were not elevated, except for those obtained in close proximity to one of the hazardous waste sites, and no association was found between serum PCB levels and exposure through these media or through occupation. PMID- 14757380 TI - Meta-analysis of feeding trials investigating cadmium accumulation in the livers and kidneys of sheep. AB - Human cadmium intake derives mainly from food sources, and cadmium can be present in high concentrations in some offal. A meta-analysis using random effects modeling was carried out to integrate the results of 21 controlled randomized trials in which sheep were fed diets with elevated cadmium levels and cadmium concentrations in their livers and kidneys were recorded after slaughter. Resulting predictions of cadmium accumulation in sheep are applicable to a broad set of exposure situations allowing the critical examination of cadmium in the human food chain. The product of the cadmium concentration in the feed and the duration of exposure to that feed were significant predictors of the cadmium concentration in livers and kidneys. The predominantly organic rather than inorganic form of cadmium in the feed further increased accumulation. Other variables (dry matter intake, the vehicle of the elevated cadmium in the diet, animal age, weight, and sex) were not significant. As a result, the prime measure to decrease the risk of cadmium from animal origin adversely affecting human health should be restricting the animals' cumulative cadmium intake. It is suggested that this might be achieved by preventing the livers and kidneys of older animals from entering the human food chain. PMID- 14757381 TI - Evaluation of metallothionein as a biomarker of single and combined Cd/Cu exposure in Dreissena polymorpha. AB - The effects of metal mixture (Cd+Cu) versus single-metal exposure on total MT response and bioaccumulation were investigated in the freshwater bivalve Dreissena polymorpha. A two-month exposure period, including two levels of contamination, was chosen for each of the two metals: 5, 10 microg/L for Cu, and 2, 20 microg/L for Cd, with mixtures of, respectively, 5 microg/L Cu+2 microg/L Cd, 5 microg/L Cu+20 microg/L Cd, 10 microg/L Cu+2 microg/L Cd, and 10 microg/L Cu+20 microg/L Cd. Total MT contents were assessed by an Ag-saturation method, and metals contents were determined by atomic absorption spectrometry. Results at the whole-organism level showed a significant and early increase of total MT biosynthesis after exposure to Cd. This increase was significantly correlated with Cd bioaccumulation. By contrast, Cu did not modify total MT response, and mussels limited Cu bioaccumulation. The mixture either did not influence or only weakly influenced metal accumulation and MT response to Cu and Cd after long-term exposure. Our results suggest that the form of MT existing in D. polymorpha was not Cu-inducible. This could limit the use of MT in D. polymorpha as a biomarker of heavy metal pollution in freshwater ecosystems. PMID- 14757382 TI - Altered brain activity in brevetoxin-exposed bluegill, Lepomis macrochirus, visualized using in vivo 14C 2-deoxyglucose labeling. AB - This study investigated the neurological effects of sublethal brevetoxin (PbTx-2) exposure in bluegill (Lepomis macrochirus) by measuring alterations in 2 deoxyglucose (2-DG) uptake in the brains of exposed fish. Changes in regional brain activity were quantified using digitized autoradiographs from exposed and control fish. Brains of brevetoxin-exposed fish had significantly higher labeling of 2-DG than brains of control fish. Regional increases in labeling were observed in the optic lobes, telencephalon, and cerebrum of PbTx-2 exposed fish. From these observations, we conclude that sublethal brevetoxin exposure in vivo in bluegill increases neurological stimulation, measured through quantification of [14C]2-DG uptake in the brain. Increases in the uptake of [14C]2-DG from this study may be indicative of differences in neural activity in the PbTx-exposed fish and are likely associated with the action of PbTx-2 on voltage-gated sodium channels (VGSC), as well as neurological alterations in calcium and neurotransmitter release downstream resulting from VGSC activation. These techniques quantify physiological alterations in fish brain activity resulting from exposure to brevetoxin and possibly other harmful algal bloom toxins. PMID- 14757383 TI - Assessment of the aquatic and terrestrial toxicity of five biodegradable polymers. AB - Radiofrequency countermeasures (i.e., chaff) may be released by fighter jets during tactical countermeasures training. Chaff cartridges, pistons, and endcaps (i.e., chaff dispenser materials), all currently made of styrene, are also released into the environment. Accumulation of chaff dispenser materials in the environment is a concern of the Department of Defense. The US Navy is exploring the possibility of constructing degradable chaff dispenser components made of biodegradable polymers. Five polymers are being considered. Degradability and toxicity tests are two of several criteria being used to evaluate various available biodegradable options. Dissolution products from four of five polymers being considered were toxic to aquatic organisms with LC50s/LOELs ranging between 1.24 and 731.30 mg total organic concentration/L. Supernatant from dissolving a 90:10 polyester amide/polyvinyl alcohol copolymer in water for 24h inhibited shoot growth of Brassica rappa and Lepidium sativum. Since our results were obtained using fractions of saturated degradable polymer solutions (1 or 10 g/L), we conclude that the tested degradable polymers were of low toxicity to the seven aquatic organisms and two terrestrial plant species used in our assays. However, our characterization of the toxicity of these degradable polymers may not be applicable to all species or environmental situations. Information gained from these studies will be used for making decisions on which polymers should be used in the engineering of environmentally friendly chaff dispenser cartridges, pistons, and endcaps. PMID- 14757384 TI - Biomonitoring of Gulf of Patras, N. Peloponnesus, Greece. Application of a biomarker suite including evaluation of translation efficiency in Mytilus galloprovincialis cells. AB - Specimens of Mytilus galloprovincialis were placed in bow nets and immersed at 3 10 m depth in a clean coastal region (reference area), Itea, and two marine stations along Gulf of Patras, N. Peloponnesus, Greece. One site is near the estuaries of the Glafkos River, which are influenced by local industrial and urban sources (Station 1); the second site, Agios Vasilios, has no evident organic pollution but is enriched in metals, particularly zinc (Station 2). One month after immersion, digestive glands were removed from the mussels and tested for lysosomal membrane stability, metallothionein content, and translational efficiency of ribosomes. In addition, gill cells were isolated and their micronuclei content was determined. Compared with the reference samples, mussels transplanted to Gulf of Patras showed a significant increased lysosomal membrane permeability and metallothionein content, reduced polysome levels, and increased chromosomal damage in relation to the contamination burden of each sampling area. Also, runoff ribosomes from mussels transplanted to Gulf of Patras (that is, ribosomes stripped of endogenous messengers and peptidyl- or/and aminoacyl-tRNAs) were less efficient at initiating protein synthesis in an in vitro-translation system than those prepared from reference samples. The whole set of data suggests that the degree of Gulf of Patras pollution differs between different sites and depends on the proximity of urban sewage and industrial outfalls. In addition, our results emphasize the importance of protein synthesis regulation as a component of the cellular stress response. PMID- 14757385 TI - Cytotoxicity and mode of action of maleic hydrazide in root tips of Allium cepa L. AB - Maleic hydrazide (MH) is an herbicide and is a regulator of the growth of buds in vegetables during storage. It is used in agriculture-in despite its known effect as a mutagenic and clastogenic agent. In this research the effect of MH on the root tips of Allium cepa L. was determined; correlations between the effects of different concentrations and exposure times on the mitotic index (MI) and induction of chromosomal aberrations (ChA) were also examined. Experiments were carried out in triplicate, using aqueous solutions of MH to concentrations of 10( 6), 10(-5), 10(-4) and 10(-3)M, at intervals of 0, 4, 8, 12, 24, and 48 h, with a control for each combination (with the MH substituted by distilled water). The results revealed an inhibition of the MI linked to the concentration and time of treatment (F=845.51, P<0.01 and F=427.58, P<0.01, respectively). For all the concentrations used and exposure periods longer than 12 h, different types of ChA were present, with significantly increased frequencies with increases in the concentration and time of exposure (P<0.01). To determine the mechanism through which the herbicide exerts its toxicity, ultrastructural electron microscopy was conducted. The results reveal nucleolar alterations, suggesting an inhibitory effect of biosynthetic activity. PMID- 14757386 TI - Clinical nutrition: new look, and ... better performance? PMID- 14757387 TI - Oxidative stress in childhood--in health and disease states. AB - The accumulated information concerning the involvement of reactive oxygen species in many clinical disorders and disease states has led to the potential for intervention with antioxidants in these cases. There are currently numerous clinical trials involving administration of antioxidants in a variety of conditions such as coronary heart disease, cataract, cancer and neurodegenerative diseases. At the same time therapeutic trials aimed at preventing and delaying the aging process are also under investigation. Numerous disorders, in childhood, have also been linked to oxidative damage. The aim of this review is to provide an overview of oxidative stress, its mechanisms, targets and damage incurred, as pertaining specifically to clinical disorders during childhood. The defense mechanisms against oxidation; the enzymatic antioxidants and low molecular weight antioxidants are defined and a number of methods commonly used for evaluation of oxidative stress (methods for measurement of lipid and protein oxidation end products and methods for measurement of antioxidant defense capacity) described. Specific diseases related to oxidative stress in infancy and childhood are reviewed and the possible effect of nutritional intake on oxidative stress in the healthy child discussed. Other issues addressed include the ability of oxidative stress, as measured in plasma to reflect intratissue oxidation, the need for a simple laboratory method for characterization of an oxidative stress 'profile', the proposed role of oxidative stress in biological processes pertaining to growth and maturation and possible implications of unrestricted antioxidant supplementation. PMID- 14757388 TI - L-alanyl-L-glutamine-supplemented parenteral nutrition improves infectious morbidity in secondary peritonitis. AB - BACKGROUND & AIMS: A growing number of randomized clinical trials suggest that glutamine (Gln) supplementation may be beneficial in a selected group of patients and conditions. However, the effects of Gln-enriched total parenteral nutrition (TPN) on recovery from acute intra-abdominal infection have not been thoroughly investigated. Therefore, the aim of this study was to investigate whether the provision of Gln-enriched TPN after surgical and medical treatment of secondary peritonitis improves infectious morbidity. METHODS: Thirty-three patients with secondary peritonitis were randomly assigned to receive either standard (n=16) TPN or L-alanyl-L-glutamine-supplemented (n=17) TPN, after medical and surgical treatment of the infectious focus. The two TPN formulae were isonitrogenous and isocaloric, which commenced the morning after surgery and ran continuously for 10 consecutive days. The control group received standard TPN, while the treatment group was given L-alanyl-L-glutamine, 0.40 g/kg/d (Dipeptiven, Fresenius Kabi, Bad Homburg, Germany). Infectious morbidity, nitrogen balance, leukocytes, lymphocytes, subpopulations CD(4) and CD(8), Immunoglobulin A (IgA), total proteins, albumin, hospital and intensive care unit (ICU) stays, and mortality were evaluated. Statistical analysis included one-way ANOVA, the unpaired Student's t-test, the Mann-Whitney U-test, chi(2) test, or Fisher's exact test. RESULTS: Patients in both groups were comparable prior to the operation. Nitrogen balance and the levels of albumin and IgA were significantly better than those in the control group. Also, a significant reduction in the infectious morbidity was found in the Gln-treated group. Lymphocyte counts as well as subpopulations CD(4) and CD(8), and proteins showed a propensity to improvement and a tendency to reduced rates of mortality were observed when comparing the groups. Hospital and ICU stays were similar. CONCLUSION: L-alanyl-L-glutamine-supplemented TPN improved the infectious morbidity of patients with secondary peritonitis. Gln supplementation to parenteral nutrition may be an alternative for enhancing host defenses and improving infectious morbidity. PMID- 14757389 TI - Levels of S100B protein are higher in mature human milk than in colostrum and milk-formulae milks. AB - BACKGROUND & AIMS: Human milk is believed to contain biological factors involved in the regulation of newborn growth, including brain development. Recently, it has also been shown to contain the calcium-binding S100B protein, regarded as a neurotrophic factor. The present study investigates the concentrations of this protein in colostrum, human milk at different levels of maturation and in milk formulae. METHODS: Samples for S100B measurements were collected from human colostrum (on day 1 after birth), from transition milk (on post-delivery days 7 and 14) and from mature milk (on day 30 after delivery) in 14 healthy women and from 14 milk-formulae. The S100B protein levels were measured using a commercially available specific immunoluminometric assay. RESULTS: Mean S100B protein levels were significantly higher in mature human milk (117.9+/-36.7 microg/l) than in transition milk at 14 days (106.7+/-38.1 microg/l) and at 7 days (92.7+/-37.8 microg/l), colostrum (74.6+/-37.6 microg/l) or milk-formulae (24.8+/-19.5 microg/l) (P<0.001, for all). A correlation between human milk S100B levels and the gestational age at which samples were obtained was also found (r=0.39; P<0.01). CONCLUSIONS: These findings, possibly related to S100B's neurotrophic role, offers useful information to the investigation of the role of S100B protein in brain maturation. PMID- 14757390 TI - Serum carnitine concentrations correlated to clinical outcome parameters in chronic hemodialysis patients. AB - Carnitine metabolism and the therapeutic use of carnitine has been a major area of interest in dialysis patients. The purpose of this study was to determine whether any correlations exist between carnitine status and selected clinical parameters in hemodialysis (HD) patients. This study was an observational study of data from patients receiving HD at a Midwest dialysis center. The subjects (n=49) were 60+/-16 (mean+/-SD) years of age and 48% male. Fifteen percent of the subjects had type 1 diabetes mellitus (DM), 29% had type 2 DM, and 25% had left ventricular hypertrophy (LVH). The serum-free and total carnitine, and acylcarnitine concentrations were: 40.3+11.8 microm/l, 22.8+/-7.3, and 17.5+/-5.9 microm/l, respectively. The serum acylcarnitine to free carnitine ratio (A/F) was 0.80+/-0.27. Blood urea nitrogen (BUN), parathyroid hormone and ejection fraction were positively correlated and age and left atrial dilation (cm) were negatively correlated with serum total carnitine (P<0.05). BUN and hematocrit were positively correlated (P<0.05) and age was negatively correlated with free carnitine. Subjects who used mannitol or were male had significantly higher concentrations of both free and total carnitine, respectively (P<0.05). Subjects using aspirin had lower concentrations of serum total carnitine (P<0.10). These results suggest certain subgroups of patients may need to be targeted for further studies with carnitine replacement therapy, i.e. long-term patients, older patients, patients with left verticular hypertrophy and left atrial enlargement, females and patients on aspirin therapy. PMID- 14757391 TI - Experience of post-pyloric feeding in seriously ill patients in clinical practice. AB - BACKGROUND: Maintaining nutrition is an integral part of patient care and when it is possible enteral nutrition is regarded as superior to parenteral nutrition. Post-pyloric feeding may enable enteral feeding to be maintained in patients who cannot tolerate nasogastric feeding. The success of post-pyloric feeding in routine clinical practice is uncertain. METHODS: One hundred and forty six consecutive patients who had 150 separate episodes of post-pyloric feeding were identified. Casenotes were reviewed to assess indication for post-pyloric feeding, prior use of alternative methods of feeding, success of achieving nutritional requirements and patient outcome. RESULTS: A post-pyloric tube was successfully placed in 138 (92%) and nutritional requirements were met by post pyloric feeding alone in 124 (83%). Post-pyloric feeding was used for between 2 and 254 days (median 14 days). Conditions for which post-pyloric feeding was used to administer nutritional support included burn injury, pancreatitis, sepsis, post-operative gastric stasis, bone marrow transplantation and chemotherapy induced vomiting. Fifty (33%) patients had an attempt at nasogastric feeding and 33 (22%) were on total parenteral nutrition before post-pyloric feeding was commenced. There was one major complication of a jejunal ulcer bleed in the series. Minor complications included displacement of the nasojejunal tube and failure to absorb feed related to gastrointestinal dysfunction. CONCLUSIONS: Post pyloric feeding can be successfully used to maintain enteral nutrition in patients who would otherwise require parenteral nutrition. PMID- 14757392 TI - Exogenous nucleosides modulate the expression of rat liver extracellular matrix genes in single cultures of primary hepatocytes and a liver stellate cell line and in their co-culture. AB - BACKGROUND & AIMS: We have previously reported the antifibrotic effect of dietary nucleotides in cirrhotic rats. In this work, we used primary rat hepatocytes, a liver stellate cell line (CFSC-2G) and co-cultures of both cell types to investigate the effects of exogenous nucleosides on the gene expression of various extracellular matrix components and on markers of liver function, and to ascertain whether the effects found in vivo are due to CFSC-2G, hepatocytes, or are the consequence of cell-cell interactions. RESULTS: Nucleosides enhanced fibronectin, laminin, and alpha1(I) procollagen levels in CFSC-2G and hepatocytes, as well as collagen synthesis and secretion in CFSC-2G. In contrast, nucleosides lowered fibronectin, laminin and alpha1(I) procollagen levels, and decreased collagen synthesis in co-cultures. Matrix metalloproteinase-13 content and collagen secretion increased in co-cultures incubated with nucleosides. Albumin increased in hepatocytes and co-cultures incubated in the presence of nucleosides. CONCLUSIONS: Nucleosides modulate the production of extracellular matrix in single cultures of hepatocytes and of CFSC-2G, and in co-cultures. This effect seems to be regulated at the translational level. The opposite behavior of single cultures and co-cultures is probably due to the fact that the latter model reproduces many of the physical and functional relationships observed in vivo between hepatocytes and stellate cells. PMID- 14757393 TI - Nasojejunal feeding in hyperemesis gravidarum--a preliminary study. AB - Hyperemesis gravidarum is a severe form of nausea and vomiting during the first trimester of pregnancy. Our objective was to assess the feasibility of nasojejunal feeding in our patients. Eleven pregnant women aged 23-46 years with hyperemesis gravidarum, persisting in spite of an in-hospital treatment of 2-15 days by intravenous fluids and antiemetic drugs and accompanied by weight loss, consented to have a nasojejunal feeding tube inserted endoscopically. Mean in hospital weight loss prior to tube insertion was 2.2+/-1.1 kg (range 0.9-5.1 kg). A clear reduction in the extent of vomiting was already apparent within the first 48 h after tube insertion, but vomiting ceased completely after a mean of 5+/-4 days (range 1-13 days). Weight gain was recorded in six patients who stayed on tube feeding for more than 4 days. Patients were encouraged to start drinking and eating along tube feeding after 3-4 days. Ceasing vomiting and a concomitant sufficient oral intake of at least 1000 kcal/day resulted in the decision to remove the tube after 4-21 days. In three cases, however, the tube was expelled by recurrent vomiting after 1-4 days, or was blocked as in one case. The tube was not reintroduced and patients did not resume vomiting. There were no complications associated with this feeding approach in this population. Only one patient was readmitted. None of the rest resumed vomiting after tube withdrawal.The above suggests that nasojejunal enteral feeding can be an effective option in hyperemesis gravidarum persisting despite intravenous fluids and antiemetic drugs. PMID- 14757394 TI - Influence of prolonged starvation on glucose kinetics in pregnant patients infected with Plasmodium falciparum. AB - Hypoglycaemia is a recognised complication of malaria in pregnancy, but its pathophysiology is not well understood. We studied the influence of fasting on glucose production and gluconeogenesis by infusion of [6,6-(2)H(2)]glucose and ingestion of (2)H(2)O in 20 female subjects, eight pregnant patients with uncomplicated falciparum malaria, six pregnant controls matched for age and trimester and six non-pregnant controls matched for age. Infection with Plasmodium falciparum induced a significant increase in glucose production (16.7+/-0.3 vs. 12.4+/-0.8 micromol/kg/min; P=0.002) and gluconeogenesis (12.5+/ 0.6 vs. 8.2+/-0.7 micromol/kg/min; P=0.001) without a change in the glucoregulatory hormone milieu, compared to the healthy pregnant controls. Extension of the fast from 20.30 to 24.30 h resulted in a rate of decline of glucose production that was similar in patients with malaria and healthy pregnant subjects, a decline that was steeper compared to the non-pregnant subjects ( 0.283 and -0.426 vs. -0.065 micromol/kg/min/h; P=0.037). The plasma glucose concentration measured at 20.30 h of fasting in the malaria patients was intermediate between the value found in the pregnant and the non-pregnant controls (4.01+/-0.2 mmol/l) while it was significantly lower in the non-infected pregnant women compared to non-pregnant controls (3.59+/-0.14 vs. 4.70+/-0.29 mmol/l; P=0.009). Plasma glucose concentration declined at a similar rate in patients with malaria and pregnant controls but faster compared to the non pregnant controls (-0.078 and -0.093 vs. -0.044 mmol/l/h; P < 0.05). We conclude that fasting is a major risk factor for hypoglycaemia in pregnancy. Non-severe Plasmodium falciparum infection in pregnant women results in higher glucose production and higher glucose levels, thereby, compared to healthy pregnant patients, delaying of the occurrence of hypoglycaemia due to fasting. The exact mechanism of hypoglycaemia in fasting pregnant women remains to be elucidated. PMID- 14757395 TI - Does additional feeding support provided by health care assistants improve nutritional status and outcome in acutely ill older in-patients?--a randomised control trial. AB - BACKGROUND & AIMS: Malnutrition is common in the elderly and increases morbidity and mortality. Most attempts to reverse malnutrition have used liquid supplements, but the findings are inconsistent. This study tests a new approach using a randomised-controlled design. The aim was to examine whether health care assistants, trained to provide additional support with feeding, can improve acutely ill elderly in-patients' clinical outcomes. METHODS: The study was carried out on three acute medicine for the elderly wards at Hammersmith Hospitals NHS Trust, London. In all, 592 patients, all over 65 years old, were recruited. RESULTS: The results showed that the median time patients received feeding support was 16 days, and the assisted group was given less intravenous antibiotics (P=0.007). However, the groups did not differ in markers of nutritional status, Barthel score, grip strength, length of stay or mortality. CONCLUSION: It was concluded that the use of health care assistants in this specialised role, in an acute setting, without change to the food provision or without targeting higher risk patients, reduced the need for intravenous antibiotics. However, the intervention did not improve nutritional status or have an effect on length of stay in the time span studied. The results highlight the difficulties of improving the intake of acutely ill elderly patients during a hospital stay. PMID- 14757396 TI - Aging, physical activity and height-normalized body composition parameters. AB - BACKGROUND & AIM: Regular physical activity prevents or limits weight gain, and gain in body mass index (BMI) and decreases mortality. The aims of the study in healthy adults were to determine the differences in fat-free mass index (FFMI) (kg/m(2)) and body fat mass index (BFMI) between age groups and determine the association between physical activity and FFMI and BFMI. METHODS: Caucasian men (n=3549) and women (n=3184) between 18 and 98 years, were classified as either sedentary or physically active (at least 3h per week at moderate or high intensity level activity). FFMI and BFMI were measured by 50 kHz bioelectrical impedance analysis. RESULTS: BFMI was significantly higher (P<0.05) in sedentary than physically active subjects and the differences became progressively greater with age. The physically active subjects were significantly less likely to have a low or high FFMI (logistic regression, P<0.001), and a high or very high BFMI (P<0.001), and more likely to have low BFMI (P<0.001) compared to sedentary adults. In contrast with fat-free mass, which was lower in older subjects, the height-normalized FFMI was stable with age until 74 years and lower thereafter. Significantly higher BFMIs were noted in older subjects. CONCLUSION: Physically active subjects are less likely to have low or high FFMI, and high or very high BFMI, and more likely to have low BFMI. In contrast to common claim that fat-free mass decreases with age, we found that FFMI was stable until 74 years. The use of FFMI and BFMI permits comparison of subjects with different heights and age. PMID- 14757397 TI - Bone quantitative ultrasound and nutritional status in severely handicapped institutionalized children and adolescents. AB - BACKGROUND & AIMS: Children with cerebral palsy (CP) have a high prevalence of pathologic fractures. Bone quantitative ultrasonography (QUS) has emerged as a radiation-free method for the assessment of bone quality and fracture risk. In this study, we applied QUS technique in order to investigate bone status in handicapped institutionalized children and adolescents. METHODS: This cross sectional study included 87 handicapped institutionalized patients. Measurements of the velocity of ultrasound wave, speed of sound (SOS), at distal radius and midshaft tibia, were performed using Omnisense 7000S analyser (Sunlight Ltd., Tel Aviv, Israel). In addition, all the participants had a thorough evaluation of nutritional status, demographic and clinical characteristics. RESULTS: Forty-five of patients had either radius or tibia bone SOS lower than -1 SD, and 21% had either radius or tibia bone SOS lower than -2.5 SD. Using step-wise regression analysis, female gender (P=0.003) and stature (P=0.008) were correlated with radius SOS. Age (P=0.03) and fracture history (P=0.04) were negatively correlated with tibia SOS. CONCLUSION: In this group of children and adolescents with CP one fifth had poor bone status as suggested by low tibia/radius SOS assessed by QUS. Female gender, stature, age and fracture history were significantly correlated with poor bone status. PMID- 14757398 TI - Physical activity cost in children following an acquired brain injury--a comparative study. AB - BACKGROUND: Alterations in energy expenditure during activity post head injury has not been investigated due primarily to the difficulty of measurement. OBJECTIVE: The aim of this study was to compare energy expenditure during activity and body composition of children following acquired brain injury (ABI) with data from a group of normal controls. DESIGN: Energy expenditure was measured using the Cosmed K4b(2) in a group of 15 children with ABI and a group of 67 normal children during rest and when walking and running. Mean number of steps taken per 3 min run was also recorded and body composition was measured. RESULTS: The energy expended during walking was not significantly different between both groups. A significant difference was found between the two groups in the energy expended during running and also for the number of steps taken as children with ABI took significantly less steps than the normal controls during a 3 min run. CONCLUSIONS: Children with ABI exert more energy per activity than healthy controls when controlled for velocity or distance. However, they expend less energy to walk and run when they are free to choose their own desirable, comfortable pace than normal controls. PMID- 14757399 TI - Continuous nasogastric tube feeding: monitoring by combined use of refractometry and traditional gastric residual volumes. AB - BACKGROUND & AIMS: Traditional use of gastric residual volumes (GRVs) is insensitive and cannot distinguish retained enteral formula from the large volume of endogenous secretions. We designed this prospective study to determine whether refractometry and Brix value (BV) measurements could be used to monitor gastric emptying and tolerance in patients receiving continuous enteral feeding. METHODS: Thirty-six patients on continuous nasogastric tube feeding were divided into two groups; patients with lower GRVs (<75 ml) in Group 1, patients with higher GRVs (>75 ml) in Group 2. Upon entry, all gastric contents were aspirated, the volume was recorded (Asp GRV), BV measurements were made by refractometry, and then the contents were reinstilled but diluted with 30 ml additional water. Finally, a small amount was reaspirated and repeat BV measurements were made. Three hours later, the entire procedure was repeated a second time. The BV ratio, calculated (Cal) GRV, and volume of formula remaining were calculated by derived equations. RESULTS: Mean BV ratios were significantly higher for those patients in Group 2 compared to those in Group 1. All but one of the 22 patients (95%) in Group 1 had a volume of formula remaining in the stomach estimated on both measurements to be less than the hourly infusion rate (all these patients had BV ratios <70%). In contrast, six of the 14 patients in Group 2 (43%) on both measurements were estimated to have volumes of formula remaining that were greater than the hourly infusion rate (all these patients had BV ratios >70%). Three of the Group 2 patients (21%) whose initial measurement showed evidence for retention of formula, improved on repeat follow-up measurement assuring adequate gastric emptying. The remaining five patients from Group 2 (35%) had a volume of formula remaining that was less than the hourly infusion rate on both measurements. The pattern of Asp GRVs and serial pre- and post-dilution BVs failed to differentiate these patients in Group 2 with potential emptying problems from those with sufficient gastric emptying. CONCLUSIONS: Refractometry and measurement of the BV may improve the clinical utilization of GRVs, by its ability to identify the component of formula within gastric contents and track changes in that component related to gastric emptying. PMID- 14757400 TI - Survey on the current practice of nutritional therapy in Portugal. AB - BACKGROUND & AIMS: This survey aimed to provide a nationwide overview about the current practice of nutrition. METHODS: Questionnaires designed by the Portuguese Association of Parenteral Enteral Nutrition were sent, with postage-paid addressee envelopes, to all hospitals and primary-care institutions. RESULTS: Hospitals' response rate was 44/100 (44%), reaching 274/359 (60%) in primary care, P=0.02. A Nutrition Support Team (NST) was reported in 34% hospitals, 40% of which were teaching hospitals. In 3/15 (9%) hospitals, NST nutrition prescriptions covered the whole hospital population; in 16% the NST only acted on a consultant basis; in 30% they were involved in purchasing nutrition products and in 18% the NST promoted teaching/training. Physicians alone prescribed oral, enteral and parenteral nutrition in 50%, 64% and 74% hospitals, respectively, and also monitored parenteral nutrition in 69% hospitals; monitoring oral/enteral nutrition involved dietitians in 46% and 41% hospitals, respectively; nurses were never involved in NSTs and seldom participated in nutritional management. Most primary-care institutions with hospital units (91%) provided nutrition therapy and nurses were then the most involved professionals, P=0.001, even prescribing regimens while physicians favoured monitoring. CONCLUSIONS: Nutrition therapy is mostly unidisciplinary; the concept of a NST and its roles is wrongly perceived; education and training are eagerly awaited. PMID- 14757401 TI - Quality of life is a primary end-point in clinical settings. AB - The objective of this review is to present and discuss the quality of life (QOL) construct, more specifically the QOL in the field of health and disease also designated as health-related quality of life (HRQOL). QOL is an everyday language concept with a relatively short history in the health field. It became a principal end-point in health care as a consequence of the development of patients' rights movements. It is important for clinical, economic and political decisions. There is no gold standard way to measure QOL and the existence of a huge number of measures and related QOL concepts makes it difficult to discuss QOL. This means that many times we are using the same expression "QOL" but we are not talking about the same thing. So we submit that it is important to keep looking for the good construct and the good measure. The reason why we decide to evaluate QOL influences the measures we choose. In general, QOL measures are based on questionnaires that must be short and easy to answer. The interest in these kind of soft measures (in opposition to the traditional hard physiologically or biochemically oriented measures) is growing fast. PMID- 14757402 TI - On the ESPEN guidelines for nutritional screening 2002. PMID- 14757403 TI - Effects of the September 28 2003 blackout in Italy in patients on home parenteral nutrition (HPN). PMID- 14757404 TI - Stem cell plasticity: an overview. AB - The capacity of adult bone marrow cells to convert to cells of other tissues, referred to by many as stem cell plasticity, was the focus of the meeting in Providence entitled "Challenges in the Era of Stem Cell Plasticity". The meeting provided a showcase for the many impressive positive results on tissue restoration including the capacity of purified marrow stem cells to restore heart, skin, and liver function in impaired mice or humans. This area of research has become a center of controversy, although it is not clear why. Calls for clonality, robustness, and function have been shown to be erroneous or premature. A call for clonality (which has been shown nicely in one study) is meaningless on a predefined stem cell population which is intrinsically heterogeneous, as they all are. Robustness means nothing; it all depends on the details of the situation. Function on an organ level is, of course, the goal of many investigators and should not be raised as a limiting consideration. Lastly, fusion has been highlighted as undermining studies with adult stem cells. It, of course, does not. Fusion is simply a means to a final goal, which occurs in certain settings of marrow conversions (transdifferentiation) and not in others. We hypothesize that the conversion phenomena may, in fact, be due to one or several marrow stem cells with broad differentiation potential which can be expressed when the cell is placed in an environment with the appropriate inductive signals. Furthermore, initial events may be relatively rare and significant conversion numbers may be obtained with massive or ongoing selection. Fusion appears in an initial mechanism in some cases and not in others. Overall, the therapeutic potential of adult marrow stem cells is very intriguing, and successful use therapeutically will probably depend on definition of the most appropriate transplant model and tissue injury. PMID- 14757405 TI - Hematopoietic differentiation of rhesus monkey embryonic stem cells. AB - Several lines of embryonic stem cells (ESC) have been established from rhesus monkey blastocysts. We have examined two of these cell lines for their potential for generating hematopoietic progenitors in cell culture, and we identified culture conditions, including supplementation with bone morphogenetic proteins (BMP), that result in hematopoietic differentiation of rhesus ESC with high efficiency. We have also characterized the resulting hematopoietic progenitor cells for their patterns of gene expression, as compared to those of hematopoietic progenitor cells harvested from rhesus monkey bone marrow. Of more than 60 genes examined in this manner, CD34+/CD38- cells derived from embryonic stem cells and those obtained from bone marrow demonstrated very similar patterns of gene expression. However, with integrin alphaL, IL-6 receptor, and flt-3 gene expression was greatly diminished or absent in CD34+/CD38- cells derived from the ESC, whereas the bone marrow-derived progenitors showed substantial expression of all of these genes. When the same type of comparison was done with mouse (D3 and CCE) as well as human (H1) embryonic stem cells, in each case comparing ESC derived hematopoietic progenitors with those harvested from bone marrow, the only consistent deficiency of gene expression was that of flt-3. In hematopoietic precursors derived from mouse ESC, globin-gene expression has previously been shown to be a useful index of the embryological maturity of the cells, and we also examined globin-gene expression in rhesus monkey ESC-derived hematopoietic precursor cells, using a semiquantitative technique. CD34+/CD38- cells demonstrated expression of the epsilon- and gamma-globin genes, but negligible levels of beta globin, suggesting that these cells were at the developmental stage in which the yolk sac and fetal liver are the primary sites of hematopoiesis. PMID- 14757406 TI - Phylogenetic aspects of tissue regeneration: role of stem cells. A concise overview. AB - Many different forms of regeneration are known among the members of the animal kingdom. Invertebrates commonly generate new individuals by sprouting and splitting off of body parts, so that the processes of asexual reproduction and of regeneration as a response to injury can hardly be distinguished. Among the adult vertebrates, regeneration of lost body parts has become exceptional rather than the rule. However, the capacity for regeneration of tissues and for the remodeling of injured organs is much better preserved than is generally appreciated. The main reason for this misconception is the enormous variety of mechanisms for replenishment, repair, and remodeling that coexist in one and the same animal. In addition, there is a considerable variation in the response to damage inflicted by different forms of injury. Our conceptions of wound repair and tissue regeneration have been radically changed by the recent discoveries of stem cell plasticity and of dedifferentiation of supposed end cells in mice and men. Seemingly, these mechanisms should provide our bodies with near inexhaustible powers of regeneration. Yet, failures to achieve just that are among the most pressing problems of modern medicine. Apparently, counterforces have evolved that prevent the orderly replacement of lost cells. The present challenge for regenerative medicine is to unravel those barriers and find means to overcome them. PMID- 14757407 TI - Understanding cell lineages as complex adaptive systems. AB - Stem cells may be considered complex reactive systems because of their vast number in a living system, their reactive nature, and the influence of local environmental factors (such as the state of neighboring cells, tissue matrix, stem cell physiological processes) on their behavior. In such systems, emergent global behavior arises through the multitude of local interactions among the cell agents. Approaching hematopoietic and other stem cell lineages from this perspective have critical ramifications on current thinking relating to the plasticity of these lineage systems, the modeling of stem cell systems, and the interpretation of clinical data regarding many diseases within such models. PMID- 14757408 TI - The potential of bone marrow cells to orchestrate homeostasis and healing in skin. AB - Bone marrow derived cells play a major role in the maintenance and repair of many tissues, mostly by the delivering of a variety of inflammatory cells. Recent evidence however suggests that bone marrow may also supply cellular substrate for rebuilding non-hematopoietic tissues following injury. This discussion addresses the potential role of bone marrow to provide cells to the skin for rebuilding of skin structures following injury and for skin maintenance. PMID- 14757409 TI - Establishing reliable criteria for isolating myogenic cell fractions with stem cell properties and enhanced regenerative capacity. AB - Despite a focused effort within the myogenic cell transplantation community, little progress has been made toward the reliable identification and isolation of progenitors that are capable of tolerating the initial posttransplantation environment and effectively regenerating clinically relevant quantities of muscle. The future success of myogenic-based treatment modalities requires an enhanced understanding of the highly heterogeneous nature of the myogenic progenitor cell pool, which has been previously documented by numerous researchers. Further, for translation of experimental animal results to clinical application, reliable in vitro selection criteria must be established and must be translatable across species. While research into the utility of surface markers is ongoing, as an alternative we have investigated in vitro cell behavioral characteristics under imposed conditions which challenge the propensity of myogenic progenitors to choose between various cell fates (i.e., proliferation, quiescence, or differentiation). Previous observations in the mouse suggest an enhanced in vivo regenerative capacity of myogenic populations with respect to their in vitro ability to maintain a proliferative and undifferentiated state [J. Cell Sci. 115 (2002) 4361]. From these observations it is thus proposed that such behavior may represent an a priori indicator of regenerative capacity following transplantation. To challenge this proposition, a rat cell isolation and transplantation model was evaluated in an identical manner. In agreement with the results obtained from the mouse, a significant correlation between regenerative capacity and induction of differentiation was observed. These results contribute to the growing body of scientific evidence documenting the underlying behavioral differences that exist between various myogenic progenitors while also, importantly, providing evidence that such differences may significantly impact the functional capabilities of these cells posttransplantation. This information further implies that from a therapeutic standpoint isolation strategies aimed toward obtaining efficient myogenic progenitors should, in the absence of a reliable surface marker(s), focus on identifying populations displaying desirable in vitro behavior (i.e., high proliferative capacity and low induced differentiation). Incorporating such criteria into cell isolation and/or purification schemes may yield significant returns in the clinical myogenic transplantation setting. PMID- 14757410 TI - Differentiation hotspots: the deterioration of hierarchy and stochasm. AB - The conception of the present-day model of hematopoiesis was begun by the work of Professor Ernst Neumann in the 19th century when he established that immature blood cells in the bone marrow migrate out into the blood vessels. Here was the birth of the hierarchical model of hematopoiesis. Jumping 135 years into the present day, recent data suggests that the stem cell regulation is not based on the classic hierarchical model, but instead more on a functional continuum. Presumptively, chromatin remodeling with cycle transit underlies changes in gene expression. This implies that the differentiative potential of primitive stem cells should also shift with cycle transit. This model proposes a less rigid system, at least in the early stem cell and progenitor compartments in which the functional characteristics of stem cells change as they go through cycle transit. We have shown that hematopoietic stem cells reversibly shift their engraftment phenotype with cytokine induced cell cycle transit. Other shifts include adhesion protein expression, cytokine receptor expression, gene expression, and progenitor phenotype. We have also found differentiation "hotspots", culture times (reflective of cell cycle state) at which stem cell differentiation was directed toward a specific lineage. This data inaugurates the end of a pure stochastic model. This work complements existing scientific work without discounting it and adds an additional dimension of complexity (or simplicity) to the process of hematopoiesis. PMID- 14757411 TI - Tissue injury in marrow transdifferentiation. AB - Recent findings indicate that adult BM contains cells that can differentiate into mature, nonhematopoietic cells of multiple tissues including cells of the kidney, lung, liver, skin and GI tract and fibers of heart and skeletal muscle. Recently the number of these observations has substantially increased, but there is a lack of information on the mechanistic issues in stem cell plasticity. In three different models for skin, liver and skeletal muscle plasticity, we have shown that following transplantation of the marrow cells from green fluorescent protein (GFP) transgenic mice, high levels of conversion of marrow cells can be identified. Injury to the tissue was the single most important factor for this phenomenon since the incidence of marrow to other tissue conversions significantly increased after tissue injury was implemented. Our studies also demonstrate the effect of radiation on the extent of marrow conversion. PMID- 14757412 TI - Homing and conversion of murine hematopoietic stem cells to lung. AB - The hematopoietic stem cell population, lineage negative-Sca positive (HSC), displays a homing defect into bone marrow (BM) after 48-h exposure to interleukin (IL)-3, IL-6, IL-11, and steel factor [J. Hematother. Stem Cell Res. 11 (2002) 913]. Cytokine treatment of murine marrow leads to reversible alterations in adhesion protein expression, which may explain the changes in homing. We evaluated 3 h homing to nonhematopoietic organs of marrow cells exposed to cytokines for 0, 18, 24, 40 and 48 h. HSC cells from C57BL/6J mice were cultured and labeled with the cytoplasmic fluorescent dye CFSE. We found homed events from uncultured cells in spleen, liver and lung, but no events were seen in duodenum or anterior tibialis muscle. Culture in cytokines led to decreased homing to marrow at 24 and 48 h with parallel changes in spleen homing. There was little variability of homing to liver, however the number, of homed events in lung was markedly increased when 24-h cultured cells were assessed. This was approximately a 10-fold increase compared to the 0 h time point (flow cytometry). Homing was determined by evaluation of frozen section (8 microm) by fluorescent microscopy for spleen, liver, duodenum, anterior tibialis and lung. Data were confirmed by flow cytometry from each organ including marrow. These data indicate the presence of a lung homing "hotspot" at 24 h of cytokine culture; this is a time when the stem progenitors cells are in mid S-phase. Altogether these data suggest that homing of marrow cell to nonmarrow organs may fluctuate with cell cycle transit and that there is a lung homing hotspot in mid-S. PMID- 14757413 TI - Tissue-specific muscle, neural and liver stem/progenitor cells reside in the bone marrow, respond to an SDF-1 gradient and are mobilized into peripheral blood during stress and tissue injury. AB - Several reports imply that bone marrow hematopoietic stem cells transdifferentiate into tissue-specific stem cells; however, the possibility of committed tissue-specific stem cells pre-existing in the bone marrow has not been dealt with adequately. We present here an alternative explanation of the so called phenomenon of stem cell transdifferentiation. First, we postulate that tissue-committed stem/progenitor cells circulate in the peripheral blood and compete for tissue-specific niches. The circulation of these cells plays an important physiological role in maintaining a pool of stem cells in distant parts of the body and the number of these cells in peripheral blood can be increased by the administration of agents similar to those used for mobilization of hematopoietic stem cells. Second, we postulate that bone marrow tissue is a source of various stem-cell chemoattractants and survival factors and provides an environment that chemoattracts tissue-specific circulating stem/progenitor cells. In this context, we envision bone marrow as a "home" or "hide-out place" not only of hematopoietic stem cells but also of already differentiated circulating tissue specific stem/progenitors. In support of this concept, we report here that mRNA of several early markers for muscle (Myf-5, Myo-D), neural (GFAP, nestin) and liver (CK19, fetoprotein) is detectable in circulating (adherent cell-depleted) peripheral blood mononuclear cells. Moreover, using real-time RT-PCR, we found that the level of expression of these markers increases in the peripheral blood of humans and mice after mobilization by G-CSF. Furthermore, using stromal derived factor-1 (SDF-1) chemotaxis and real-time RT-PCR analysis, we present evidence that early tissue-specific stem cells reside in normal human and murine bone marrow, express the CXCR4 receptor on their surface and can be highly enriched (in humans and mice) after chemotaxis to SDF-1 gradient. All our experiments were performed on freshly isolated cells to exclude the potential contribution of transdifferentiated hematopoietic stem or mesenchymal cells in the culture. We maintain that any transdifferentiation studies employing populations of bone marrow cells should rule out the possibility that the apparently pure hematopoietic stem cell population could in fact contain pre existing tissue-specific stem/progenitors. PMID- 14757414 TI - The conceptual application of systems theory to stem cell biology: a matter of context. AB - Much of our current understanding of stem cell biology is derived from reductionist observations made out of context from the functional stem cell compartment. The extrapolation of observations of cellular behavior from highly contrived experimental circumstances to intact biological systems represents an unsupportable assumption on which we currently rely. The conceptual application of systems theory to stem cell biology requires a shift in our view of experimental stem cell science toward the reality of cellular function within a complex biological system. Although the complexity of the system is beyond current mathematical modeling, the conceptual framework of systems theory can provide context to multiple controversies in stem cell biology. PMID- 14757415 TI - Stem cell plasticity or fusion: two approaches to targeted cell therapy. AB - The possibility the adult stem cells may be able to differentiate along atypical developmental pathways has garnered widespread attention. Recent papers have demonstrated that some "plasticity" arises as a result of cellular fusion events. This review attempts to highlight and reconcile the current data on both sides of the plasticity debate. PMID- 14757416 TI - Applications and technical challenges of fluorescence in situ hybridization in stem cell research. AB - Stem cell research, maintenance, and manipulations have advanced significantly in recent years, and we now witness successful clinical applications of stem therapies. However, challenges in regard to karyotypic stability and the ploidy status of stem cell lines have been addressed only marginally. Our approach to develop technology to address these highly relevant issues is based on fluorescence in situ hybridization (FISH) using nonisotopically labeled DNA probes. As a single cell analysis technique, FISH is expected to be applicable to a variety of cells and tissues including interphase and metaphase cell preparations as well as tissue sections and biopsy material. Over the last decade, our laboratories generated a large number of probes and probe sets for the molecular cytogenetic analyses of stem cells derived from different species. These probes and the introduction of spectral imaging bring us close to be able to perform a comprehensive karyotype analysis of single interphase cell nuclei. It should furthermore be possible to couple cytogenetic investigations of the cellular genotype with analysis of gene expression. This report summarizes our technical achievements relevant to stem cell research and outlines plans for future research and developments. PMID- 14757417 TI - A large animal noninjury model for study of human stem cell plasticity. AB - In this paper, we describe an experimental model that allows evaluation of the full potential of stem cells under normal physiological conditions and in the absence of genetic or injury-induced dysfunction, thus serving as a valuable tool for the study of the mechanism(s) underlying stem cell differentiation. The fetal sheep model of human stem cell transplantation permits the robust formation of donor-derived tissue-specific cells in the absence of selective pressure given its unique characteristics: the preimmune status of the fetus allows donor stem cell engraftment without significant rejection and the existence of the naturally occurring migratory patterns in the fetus facilitates the widespread efficient distribution of donor stem cells throughout the body. The cells are then influenced by the stimulatory environment of the specific tissue or organ to undergo proliferation and directed differentiation. The versatility of this noninjury fetal model of human stem cell plasticity was demonstrated by revealing the differentiative potential of different populations of both human hematopoietic stem cells and human mesenchymal stem cells (MSC). PMID- 14757418 TI - Targeting of Lin-Sca+ hematopoietic stem cells with bispecific antibodies to injured myocardium. AB - Bispecific antibodies (BiAbs) are being used to target T cells or other immune cells to antigen-specific tumor targets. Anti-CD3 activated T cells (ATC) armed with anti-CD3 x anti-HER2 BiAb (HER2Bi) have been used to target Her2/neu + breast and prostate carcinoma cells. We adapted BiAb technology to target stem cells to injured myocardium. Since myocardial infarctions can lead to cardiac death and disability, rapid repair and rejuvenation of damaged myocardium is critically needed. Effective homing of stem cells and transdifferentiation of the stem cells into functional elements of the myocardium is needed for repair of damaged myocardium. We use a BiAb that binds c-kit on murine stem cells and VCAM 1 adhesion molecules up-regulated on injured myocardial cells. To test for specific binding and homing in a mouse, we produced anti-c-kit x anti-VCAM-1 to target purified Lin-Sca+ murine stem cells to the injured myocardium. Mice with infarcts created by ligation of the left anterior descending artery (LAD) were directly injected with armed stem cells or injected via the internal jugular vein (IJ) with FACS sorted Lin-Sca+ stem cells from bone marrow after fluorescent dye labeling. There were increased numbers of armed Lin-Sca+ cells retained in infracted myocardium after direct injection of armed Lin-Sca+ cells and increased numbers of Lin-Sca+ cells that were found in injured myocardium after IJ injection. These results suggest that stem cells retargeted with BiAb can be directly injected and retained by injured myocardium or targeted to injured myocardial tissues for tissue regeneration. PMID- 14757419 TI - The chronic wound: impaired healing and solutions in the context of wound bed preparation. AB - In the past few years, a different paradigm for the understanding and treatment of chronic wounds has emerged. The term used to describe this new context in which failure to heal is viewed is "wound bed preparation". This term is revolutionizing the way we approach chronic wounds, and has allowed chronic wounds to gain independence from established models of acute injury. Within the context of wound bed preparation, impaired healing and solutions to it are being addressed in novel ways. In this report, we make use of the diabetic ulcer as an example of a chronic wound, and emphasize the pathophysiological principles, the cellular and molecular abnormalities, and the solutions offered by the new approaches of gene therapy and stem cells. The emerging view is that chronic wounds are characterized by resident cells that have undergone phenotypic changes that need to be corrected for optimal healing to occur. We have established in animal models and in humans that stem cells have the potential to bring about fundamental changes in the repair process and, ultimately, a "quantum" jump in our therapeutic success. PMID- 14757420 TI - Marrow cells as progenitors of lung tissue. AB - There is accumulating evidence showing that marrow-derived cells can engraft as differentiated epithelial cells of various tissues, including the lung. These findings challenge long-held views regarding the basic biology of stem cells. Elucidating the fundamental mechanisms controlling these processes is the major challenge of this field. Regardless, these experiments suggest new strategies for the treatment of chronic diseases. PMID- 14757421 TI - Stem cells and regeneration of the cardiovascular system: facts, fictions, and uncertainties. AB - The data reviewed here support the view that bone marrow (BM) stem cells can migrate to infarcted myocardium and differentiate into mature, functional cardiomyocytes. Cytokines can mobilize BM stem cells from marrow leading to homing of progenitor cells to infarcted myocardium. Interestingly, contractile performance in these studies appears greater than the capacity of the regenerated cell mass and its biomechanial organization predict, suggesting that the function of the regenerated cell per se may not alone account for global functional recovery. PMID- 14757422 TI - Human stem cell research: some controversies in bioethics and public policy. AB - In the United States, controversy has raged about ethical public policies toward some potential sources of human stem cells for research, particularly human embryos left over from in vitro fertilization (IVF) or created through somatic cell nuclear transfer. This article critically examines the ethical and policy issues, particularly as they have emerged in the reports and recommendations of two presidentially appointed advisory bodies: The National Bioethics Advisory Commission (NBAC) and the President's Council on Bioethics (PCB). PMID- 14757423 TI - Do G-CSF mobilized, peripheral blood-derived stem cells from healthy, HLA identical donors really engraft more rapidly than do G-CSF primed, bone marrow derived stem cells? No! AB - For more than a decade, the notion that peripheral blood-derived stem cells engraft more rapidly than bone marrow-derived stem cells after high-dose therapy has dominated our thinking. Recently, reports that granulocyte colony-stimulating factor (G-CSF) induces a proteolytic marrow microenvironment have provided mechanistic support for that belief, compelling us to review our own experience of 29 consecutive transplants with HLA-identical blood and marrow stem cells. In contrast to several reported randomized controlled trials, we found marrow stem cells engraft just as rapidly (median day 11 for granulocytes over 500/microl and median day 17 for platelets over 20,000/microl) as blood stem cells (median day 12 and median day 19, respectively) if the donor is treated with G-CSF in the same manner before marrow harvest as the donor is treated with G-CSF before leukapheresis. These observations with healthy HLA-identical donors confirm the results of our prior randomized autotransplant study. We propose the concept that the level of activation of the stem cells (induced by G-CSF) determines engraftment kinetics and not the anatomical site of derivation. PMID- 14757424 TI - Distinct phenotypic expression of two de novo missense mutations affecting the dimer interface of glucose-6-phosphate dehydrogenase. AB - Mutations encoding class I glucose-6-phosphate dehydrogenase (G6PD) variants are associated with chronic nonspherocytic hemolytic anemia (CNSHA), the most severe phenotypic expression of G6PD deficiency. These mutations frequently affect the G6PD dimer interface that is essential for enzymatic activity. We detected two de novo missense mutations concerning residues located close together in the dimer interface in two patients with severe G6PD deficiency. A novel c.1225C>T missense mutation was identified in a male neonate who presented with hemolysis and severe hyperbilirubinemia and the predicted Pro409Ser substitution constituted a novel class I variant, designated G6PD Utrecht. G6PD deficiency in the second patient was due to the once previously reported class I variant G6PD Sumare (Val431Gly). Structural analysis revealed that the mutated residues Pro409 and Val431, located on different subunits, interact directly across the subunit interface and perturb formation of the G6PD dimer upon mutation. Favism and mild chronic hemolysis characterized the phenotype of the patient with G6PD Sumare which contrasts with the more severe clinical picture of the patient with G6PD Utrecht and, in addition, that of the patient originally described with G6PD Sumare. We postulate that this G6PD variant is at the crossing between class I and class II G6PD deficiency and its ultimate phenotypic expression is either aggravated or ameliorated by other (extra)genetic factors. PMID- 14757425 TI - Rare thalassemic syndrome caused by interaction of Hb Questembert (alpha1 codon 131, TCT>CCT, Ser>Pro) with an alpha-thalassemia-2 deletion: implications for diagnosis and management. AB - Abnormal globin chain biosynthesis may result in deficient quantity (thalassemia) or structural variation (abnormal hemoglobins) and traditionally, they represent two phenotypically distinct groups of disorders. However, the phenotypic expression of unstable hemoglobin variants often combine features of thalassemia together with variable peripheral hemolysis. To achieve definitive diagnosis in a child presenting with hemolytic anemia along with features associated with thalassemia intermedia, we evaluated clinical, hematological, biochemical, globin biosynthetic and molecular data. Definitive diagnosis was achieved by DNA analysis which characterized the proband to be a compound heterozygote for a common alpha-thalassemia-2 deletion (3.7 kb) and Hb Questembert (alpha131[H14] Ser>Pro) caused by a C>T mutation in codon 131 of the alpha1 globin gene in trans. The phenotype of thalassemia intermedia with marked dyserythropoiesis, found in patients inheriting alpha-thalassemia mutations along with unstable alpha-globin variants (i.e., alpha-thalassemic hemoglobinopathies), represents a distinct type of thalassemic syndrome. The proband in this study additionally had variable peripheral hemolysis, presumably related to characteristics of the unstable Hb Questembert. There is minimal experience for the management of such atypical cases and this case illustrates that it is probably insufficient to monitor clinical status in patients with such hemoglobinopathies based only on the levels of hemoglobin. PMID- 14757426 TI - A novel mutation in the glucose-6-phosphate dehydrogenase gene in a subject with chronic nonspherocytic hemolytic anemia--characterization of enzyme using yeast expression system and molecular modeling. AB - Glucose-6-phosphate dehydrogenase (G6PD) deficiency is the most common human enzymopathy. Human G6PD gene is highly polymorphic, with over 130 mutations identified, many of which cause hemolytic anemia. We studied a novel point mutation in the G6PD gene 1226 C-->G, predicting the proline 409 to arginine substitution (G6PD Suwalki). We expressed the human wild-type and mutated G6PD gene in yeast Saccharomyces cerevisiae which allowed the characterization of the Suwalki variant. We showed that human wild-type, as well as the mutated (1226 C- >G) G6PD gene, functionally complemented the phenotype displayed by the yeast strain with disruption of the ZWF1 gene (homologue of the human G6PD gene). Comparison of wild-type (wt) human G6PD purified from yeast and from blood shows no significant differences in the Km values for G6P and in the utilization rate for the substrate analogue, 2-deoxyG6P. The P409R substitution leads to drastic changes in G6PD kinetics. The specific activity as well as stability of mutated G6PD is also significantly reduced. Besides this, the effect of this mutation was analyzed using a model of the tertiary structure of the human enzyme. The localization of the P409R mutation suggests that it may influence the stability of the whole protein by changing tetramer interactions and disturbing the binding of structural NADP+. PMID- 14757427 TI - The ferroportin disease. AB - A new inherited disorder of iron metabolism, hereafter called "the ferroportin disease," is increasingly recognized worldwide. The disorder is due to pathogenic mutations in the SLC40A1 gene encoding for a main iron export protein in mammals, ferroportin1/IREG1/MTP1, and it was originally identified as an autosomal dominant form of iron overload not linked to the hemochromatosis (HFE) gene. It has distinctive clinical features such as early increase in serum ferritin in spite of low-normal transferrin saturation, progressive iron accumulation in organs, predominantly in reticuloendothelial macrophages, marginal anemia with low tolerance to phlebotomy. Ferroportin mutations have been reported in many countries regardless of ethnicity. They may lead to a loss of protein function responsible for reduced iron export from cells, particularly reticuloendothelial cells. Now, the disorder appears to be the most common cause of hereditary iron overload beyond HFE hemochromatosis. PMID- 14757428 TI - Deferiprone versus deferoxamine in patients with thalassemia major: a randomized clinical trial. PMID- 14757430 TI - GA-binding protein transcription factor: a review of GABP as an integrator of intracellular signaling and protein-protein interactions. AB - GA-binding protein (GABP) is an ets transcription factor that controls gene expression in several important biological settings. It is unique among ets factors, since the transcriptionally active complex is an obligate heterotetramer that is composed of two distinct proteins. GABPalpha includes an ets DNA binding domain (DBD), while a distinct protein, GABPbeta, contains ankyrin repeats and the transcriptional activation domain (TAD). GABP was first identified as a regulator of viral genes and nuclear respiratory factors. However, GABP is now recognized to be a key transcriptional regulator of dynamically regulated, lineage-restricted genes, especially in myeloid cells and at the neuromuscular junction. Furthermore, it regulates genes that are intimately involved in cell cycle control, protein synthesis, and cellular metabolism. GABP acts as an integrator of cellular signaling pathways by regulating key hormones and transmembrane receptors. In addition, GABP itself, is a target of phosphorylation events that lie downstream of signal transduction pathways. The physical and functional interactions of GABPalpha and GABPbeta with each other and with other transcription factors and co-activators are key to its ability to regulate gene expression. Its role in regulating genes involved in fundamental cellular processes places GABP at the nexus of key cellular pathways and functions. PMID- 14757431 TI - The interferon-inducible p200 family of proteins: a perspective on their roles in cell cycle regulation and differentiation. AB - The interferon-inducible p200 (IFI-200) family of proteins is among the numerous gene products induced by interferons (IFNs), which are important regulators of cell growth, immunomodulation and host resistance to tumors and viral infections. The members of this family of proteins are highly homologous to one another and consist of five murine proteins including p202, p203, p204 and p205 as well as three human homologues; IFI-16, myeloid cell nuclear differentiation antigen (MNDA) and absent in melanoma (AIM) 2. They possess at least one copy of a conserved 200 amino-acid motif which exists in two types; the a and b domains. Most of the IFI-200 proteins also possess a domain in apoptosis and interferon response (DAPIN)/PYRIN domain, which is a conserved motif associated with protein protein interactions in the regulation of apoptotic and inflammatory signaling pathways. The p200 proteins have been implicated in cell cycle regulation and differentiation based on their ability to interact with and modulate the activities of multiple transcriptional factors such as Rb and p53, and there are significant findings that link mutations in their genetic loci to the incidence of cancer. Here, we describe the structure and biological activities of these proteins, and discuss recent studies that describe their relevant roles in processes regulating cell proliferation and differentiation. PMID- 14757432 TI - Hematopoietic stem cells and endothelial cell precursors express Tie-2, CD31 and CD45. AB - Early theories of tumor angiogenesis suggested that preexisting vessels surrounding the tumor were the principal source of the tumor vasculature but recent evidence suggests that endothelial progenitor cells (EPC) migrate from the marrow play an important role in developing the tumor blood supply. In a mouse model, in which the vascularization of a transplantable tumor was studied after bone marrow (BM) transplantation, we show that cells that express Tie-2, Sca-1, CD31 and CD45 function as both BM EPC and primitive hematopoietic stem cells. BM cells from transgenic mice expressing green fluorescent protein (GFP) under the control of the endothelial lineage-specific Tie-2 promoter (Tie-2 /GFP) were used to reconstitute irradiated (12 Gy) wild-type mice. Five donor BM cell populations were studied: (1) whole BM; (2) Sca-1-enriched BMC; (3) GFP/Tie-2+, Sca-1+ BMC; (4) GFP/Tie-2-, Sca-1+ BMC and (5) Sca-1-depleted BMC. After 4 weeks, the mice were injected with Tg.AC tumor cells. Three weeks later, sections from the tumors were stained for CD31 and examined for Tie-2-driven GFP expression. BM-derived endothelial cells were found only in mice transplanted with bone marrow containing populations of Tie-2+, Sca-1+ cells. As few as 3500 of these cells were sufficient to radioprotect lethally irradiated mice. Thus, we conclude that a rare subset of BMC (approximately 4 x 10(-3)%) with the putative properties of hemangioblasts have an active Tie-2 promoter. Selection of Tie-2+Sca-1+ BMC enriches for marrow-derived EPCs that participate in tumor angiogenesis and cells that can provide hematopoietic reconstitution of marrow-ablated mice. PMID- 14757433 TI - A single amino acid change in the binding pocket alters specificity of an anti integrin antibody AP7.4 as revealed by its crystal structure. AB - Monoclonal antibody (mAb) AP7.4 is an anti-integrin antibody recombinantly expressed in Escherichia coli specific to alphavbeta3. It is known that in a variety of RGD-containing molecules, ligand specificity is regulated by structural determinants within the immediate vicinity of the RGD sequence. To better understand the role of the RGD sequence in integrin specificity, we report here the three-dimensional structure of Fab of mAb AP7.4 to a resolution of 2.25 A. The crystals belong to a triclinic space group P1 and the volume of the unit cell is consistent with the presence of two Fab molecules in it. The RGD sequence is located at the tip of a flexible loop in the complementary determining region (CDR-3) of the heavy chain. It has been shown that specific recognition of RGD ligands by their receptors is influenced mainly by the conformation of the tripeptide RGD and the amino acid residues flanking it on either side. Hence, the flexibility of the RGD-carrying loop observed in the crystal structure may stem from the fact that the antibody molecule mimics the function of these cell adhesion molecules. PMID- 14757434 TI - Facilitative glucose transporter gene expression in human lymphocytes, monocytes, and macrophages: a role for GLUT isoforms 1, 3, and 5 in the immune response and foam cell formation. AB - Cellular glucose uptake is mediated by a family of facilitative glucose transporters (GLUT) exhibiting differences in kinetics, substrate specificity, and tissue-specific expression. GLUT isoform expression has not been comprehensively studied in human leukocytes, which participate in immune and inflammatory responses and are critical for host defense. Therefore, we studied the regulated expression of GLUT 1-5 mRNA and protein in isolated human lymphocytes and monocytes and in human THP-1 macrophages and foam cells. Lymphocytes expressed GLUT 1 and GLUT 3 proteins, and cellular levels of both isoforms were augmented 3.5- to 6-fold following activation by phytohemagglutinin (PHA). Monocytes expressed 8.4-fold more GLUT 3 protein and 88% less GLUT 1 than lymphocytes, and activation by lipopolysaccharide (LPS) led to a 1.9-fold increase in GLUT 1. At the level of mRNA expression, GLUT 3 mRNA was the most prevalent GLUT mRNA species in monocytes, while lymphocytes expressed equal numbers of GLUT 1 and GLUT 3 transcripts. Differentiation of THP-1 monocytes into macrophages was associated with marked induction of GLUT 3 and GLUT 5 protein expression, and high levels of GLUT 1, GLUT 3, and GLUT 5 were maintained after transformation to foam cells. GLUT 5 mRNA was expressed in 2-fold greater abundance in macrophages and foam cells than that observed for GLUT 1 mRNA, while the level of GLUT 3 mRNA was intermediate. This facilitative glucose transporters are differentially expressed and regulated in human leukocytes in a pattern that could facilitate cellular functions. Speculatively, high GLUT 1 and GLUT 3 expression could provide cellular fuel for the immune response, and high levels of high-affinity GLUT 3 in macrophages might allow the cell to compete with pathogens for hexoses, even in the presence of low interstitial glucose concentrations. Ample expression of GLUT 1 and GLUT 3 in foam cells could also provide hexose substrates and promote lipid loading. The role for high levels of the fructose transporter GLUT 5 in macrophages and foam cells is unknown since interstitial and circulating fructose concentrations are low in these cells. PMID- 14757435 TI - Knockdown of prothrombin in zebrafish. AB - Thrombin is a serine protease generated from its zymogen, prothrombin, and plays a central role in the coagulation cascade. It is also important for mammalian development. The zebrafish has now been established as an excellent genetic model for studies on mammalian hemostasis and development. In this report, we used prothrombin-specific antisense morpholinos to knock down the levels of prothrombin to characterize the effects of prothrombin deficiency in the zebrafish embryo. Prothrombin morpholino-injected zebrafish embryos yielded an early phenotype exhibiting severe abnormalities that later showed occasional bleeding. In a second late phenotype, the embryos had no observable morphological abnormalities in early stages, but showed occasional bleeding at later stages. These phenotypes resembled characteristics shown by prothrombin knockout mice. Laser-induced vascular injury on some of the normal appearing phenotypic larvae showed a prolonged time to occlusion, and recombinant zebrafish prothrombin injected into these larvae restored a normal time to occlusion thus showing the specificity of the morpholino effect. The system developed here should be useful for investigation of the role of thrombin in vertebrate development. PMID- 14757436 TI - Transplanted ER-MP12hi20-58med/hi myeloid progenitors produce resident macrophages from marrow that are therapeutic for lysosomal storage disease. AB - Lysosomal storage diseases (LSD) respond to bone marrow (BM) transplantation when donor-derived cells deliver needed enzyme. Hypothetically, the ubiquitous resident macrophages (MPhi) are the primary delivery vehicle of therapeutic protein. In mucopolysaccharidosis type VII (MPS VII) mice with LSD, transplanted mature MPhi reduce undegraded glycosaminoglycans (GAG) in the lysosome but are incapable of self-renewal, leading to return of storage after 1 month. We show here that a population of early BM-derived myeloid progenitors devoid of long term hematopoietic stem cells (LT-HSC) engrafted MPS VII BM, released monocytes into peripheral blood (PBL), and engrafted tissues at known sites of resident MPhi. These primitive Mac-1- cells were sorted from normal whole BM and were defined by ER-MP12hi20-58med/hi labeling. Lysosomal storage was reduced in liver, spleen, thymus, heart, kidney, and bone. Cells persisted for 3 months, suggesting self-renewal capacity or a long half-life. Cells sorted from BM by ER-MP12-20hi marker expression (which are maturer myeloid cells that express Mac-1) engrafted tissues instead of BM and quantitatively repopulated less than cells derived from the ER-MP12hi20-58med/hi population. Also, reduction of lysosomal storage was variable and generally less when compared to that following transplantation of immature ER-MP12hi20-58med/hi cells. We conclude that primitive myeloid progenitors are more therapeutic for LSD than mature myeloid cells due to their greater longevity and increased capacity to seed tissues. The ability of cells derived from these primitive precursors to seed deep within tissues make them excellent candidates for both cellular therapy and gene transfer techniques to cure a wide range of metabolic diseases. PMID- 14757437 TI - Effect of enzyme replacement therapy on gammopathies in Gaucher disease. AB - Chronic antigenic stimulation by the abnormal lipid storage has been postulated to be the mechanism underlying anecdotal reports of monoclonal and polyclonal gammopathies as well as an increased incidence of multiple myeloma in patients with Gaucher disease of all ages. With the advent of specific enzyme therapy, it has been possible to ascertain whether signs and symptoms associated with Gaucher disease are true features of the disorder by virtue of their responsiveness to treatment. The purpose of this study was to assess the incidence of polyclonal and monoclonal gammopathies in a large cohort of patients and the effect of enzyme treatment. All adult patients whose records of immunoglobulin levels were available at presentation or at the advent of enzyme replacement therapy (ERT), and who had been followed for 2 years or receiving ERT for at least 2 years, respectively, and for whom there were also immunoglobulin levels at their most recent follow-up, were included in the study. The incidence of polyclonal gammopathies ranged between 14% and 25% among treated and untreated patients. There were statistically significant percentage decreases per year of enzyme therapy in polyclonal but not monoclonal (1% of all patients) gammopathies. Among enzyme-treated patients, there was no statistically significant difference among patients with regard to spleen status or relative to other parameters of disease severity, hepatitis status, age or gender. This study represents the largest database of gammopathies among patients with Gaucher disease from a large referral clinic. Because there was no correlation of abnormal immunoglobulin levels with disease severity, etiology may not be related to lipid accumulation per se but perhaps reflects a secondary, enzyme-sensitive process, whereas monoclonal gammopathies remain unaffected. PMID- 14757438 TI - Expression and functional characterization of mutated glucocerebrosidase alleles causing Gaucher disease in Spanish patients. AB - BACKGROUND: Gaucher disease (GD) is a heterogeneous disease characterized by an impaired activity of the lysosomal glucocerebrosidase. This heterogeneity is attributed in part to the existence of a large number of mutations in the corresponding gene. SUBJECTS AND METHODS: To establish genotype-phenotype relationships, we analyzed the residual enzyme activities of six naturally occurring mutations found in Spanish population in the glucocerebrosidase gene [c.160G > A (V15M), c.485T>C (M123T), c.914C>T (P266L), c.1124T>C (L336P), c.1207A>C (S364R) and c.1510-1512delTCT (S465del)]. The mutated genes were subcloned into the mammalian expression vector pCR 3.1 and expressed by transient transfection in COS cells. The enzymatic activity of the expressed protein were measured and compared with the wild-type human glucocerebrosidase cDNA. Also, two previously alleles, c.1226A>G (N370S) and c.1448T>C (L444P), were used for comparative purposes. RESULTS: The residual activity of the expressed proteins using the synthetic substrate (4-methylumbelliferyl-beta-D-glucopyranoside, 4MU Glu) ranged from 5.5% (for the 3-bp deletion) to 42.7% (for S364R mutation) of the activity of the wild-type enzyme. CONCLUSION: The present analyses may help to better understand the molecular basis and the pathogenesis of Gaucher disease. However, results of expression of mutated enzymes are necessary but not sufficient to explain the ultimate clinical outcome of Gaucher disease. PMID- 14757439 TI - A novel retrovirus provides the cooperating oncogenic event(s) required to demonstrate the tumor suppressor activity of p15Ink4b in myeloid cells in vivo. AB - Cancer is a multistep process resulting from an accumulation of several genetic changes. The determination of cooperating events in experimental models can help scientists decipher specific neoplastic pathways and place genes with similar functions in complementation groups. In leukemia models, retrovirus tagging is a powerful approach to determine genes that cooperate with oncogenic transgenes or tumor suppressors that have undergone targeted deletion. Experimental models for B and T cell leukemias involving transgenic c-myc were the first to show the utility of retroviral tagging. Here we review these experiments and present examples of new models of myeloid leukemia where retroviruses have collaborated with a transgene [Cbfbeta-MYH111 from Inv(16)] and with loss of a tumor suppressor (Ink4b) mice to induce disease. PMID- 14757440 TI - Incidence of BCL-2 gene rearrangements in Argentinean non-Hodgkin lymphoma patients: increased frequency of breakpoints outside of MBR and MCR. AB - The t(14;18)(q32;q21) is the most frequent cytogenetic abnormality observed in follicular lymphoma (FL), but also occurs in diffuse large B-cell lymphoma (DLBCL). We have evaluated the frequency of this translocation in 106 Argentinean non-Hodgkin lymphoma (NHL) patients: 83 with diagnosis of FL and 23 with DLBCL. Nested (N) and long-distance PCR (LD-PCR) approaches were used. By N-PCR, a total of 42 (51%) FL cases showed BCL-2 rearrangement: 28 (34%) for major breakpoint region (MBR) and 14 (17%) for minor cluster region (mcr). By LD-PCR, additional 23 (28%) new positive cases were found: 10 (12%) for MBR and 13 (16%) for mcr. These data make a total of 65 (78%) positive cases for BCL-2 gene rearrangements. In DLBCL cases, N-PCR detected two (9%) cases with the MBR breakpoint and one (4%) with mcr. Seven (30%) new positive cases by LD-PCR were found: four (17%) for MBR and three (13%) for mcr, showing a total of 10 (43%) positive cases. Thus, both FL and DLBCL had high frequencies of breakpoints located between MBR and mcr clusters. Our N-PCR results in FL (51%; 95% CI, 40-62%) showed statistical differences with respect to the pooled data from USA (P < 0.0001) and overlapped with the frequencies from Asia and Europe. In DLBCL, no significant differences with respect to the literature were found. This data support the idea that FL may be a heterogeneous malignancy with distinct molecular pathogenesis and suggest that the geographic differences may be related with the distribution of breakpoints that are widely spread throughout the sequence stretch between MBR and mcr. PMID- 14757441 TI - Protein expression of PDZ-binding kinase is up-regulated in hematologic malignancies and strongly down-regulated during terminal differentiation of HL-60 leukemic cells. AB - PBK/TOPK is a recently identified 322 amino acid serine/threonine kinase that is phosphorylated during mitosis and may include p38 MAPK among its targets. Previous work has shown up-regulated expression of PBK/TOPK mRNA in a variety of tumor cell lines and fetal tissues, suggesting a role for this kinase in malignant cell proliferation. In this paper, PBK/TOPK protein expression was examined in a variety of primary hematologic neoplasms: PBK/TOPK was readily detected in 9 of 12 AML samples (75%), in 3 of 3 ALL samples, and in 1 sample each of a plasmacytoma and blastic type mantle cell lymphoma where it was strongly expressed. In contrast, PBK/TOPK was only weakly expressed in 2 samples of G-CSF-mobilized peripheral blood stem cells that were enriched in CD34+ progenitors by immunoselection. Furthermore, when HL-60 myeloid leukemic cells were differentiated with phorbol ester (TPA), PBK/TOPK protein expression was strongly down-regulated by 24 h. Under these same conditions, phosphorylated c Myc was rapidly down-regulated (by 4 h), while the levels of cyclin D1 and phosphorylated p38 were constant. Notably, of 5 clinical samples that strongly expressed PBK/TOPK, 4 also strongly expressed phosphorylated c-Myc, while only 1 of 3 PBK/TOPK negative samples expressed phosphorylated c-Myc. These data show that PBK/TOPK protein is up-regulated in a variety of hematologic malignancies and may be involved in leukemic cell growth. Additional studies are warranted to determine if PBK/TOPK would be a valuable target for novel therapeutics. To this end, we also describe the derivation of clones of 293 (human embryonic kidney) cells, which carry an inducible kinase-defective mutant of PBK/TOPK. This model may be useful for studying the effects of down-regulated PBK/TOPK function. PMID- 14757442 TI - Familial (inherited) leukemia, lymphoma, and myeloma: an overview. AB - We have reviewed the world's literature that addresses familial leukemia, lymphoma, and myeloma. We have catalogued the phenotypic abnormalities associated with an increased risk of developing a hematological malignancy. These syndromes, such as Fanconi anemia or familial platelet syndrome, have been well characterized and in many cases the gene responsible for the predisposition has been defined. We have focused, however, on reports of a familial incidence of hematological malignancy in which no prior predisposing syndrome was reported. In this circumstance, so-called pure familial leukemia, lymphoma, or myeloma, the intergenerational incidence of disease occurred in ostensibly healthy persons. These families have been grouped into sets in which (a) anticipation, (b) immune abnormalities, (c) linkage to HLA phenotypes, (d) linkage to chromosome abnormalities, or (e) gene abnormalities have been reported. They have also been grouped by type of leukemia. Purely descriptive reports, not accompanied by some information on pathogenesis, have not been included. They are catalogued in some of the references cited in this paper. Anticipation is a prominent feature of familial leukemia, lymphoma, and myeloma, supporting the concept of germline transmission of a susceptibility gene. Although linkage to an HLA phenotype occurs in some families, no consistent intrafamilial pattern has emerged. Deletion of chromosome 7 is associated with familial acute myelogenous leukemia, but no other recurring localization has been established. Although putative susceptibility genes have been identified in some families, the likelihood is that the mode of inheritance is different in different families and different genes are involved even within a specific Mendelian pattern. Although as yet not reported, the frequency of familial CLL and the intensity of its study indicates that the gene or genes involved in that familial disorder(s) should be identified conclusively soon if sufficient families for study can be assembled through international cooperation. PMID- 14757443 TI - Interferon-alpha suppresses proliferation of chronic myelogenous leukemia cells K562 by extending cell cycle S-phase without inducing apoptosis. AB - We examined the effects of interferon-alpha (IFN-alpha) treatment on the growth, cell cycle, proliferation, and apoptotic parameters as well as adhesive properties and proteome of chronic myelogenous leukemia (CML)-derived K562 cells. IFN-alpha treatment (200 to 600 U/ml, 24 to 72 h) suppressed growth and caused accumulation of K562 cells in the S-phase of cell cycle (increase in S-phase cells by up to 52% in comparison with the untreated controls) at the expenses of cells in G1-phase. No transition of cells to G0-phase occurred as followed from Ki-67 protein determination. Although the level of chimeric gene product, BCR-ABL mRNA coding for BCR-ABL protein with anti-apoptotic properties, decreased by 30%, apoptosis was not triggered as judged from Annexin-V, APO2.7, and TUNEL assays. Adhesion of K562 cells to fibronectin-coated surfaces increased by up to 52% as determined by calcein assay. The proteomic analysis (2-D electrophoresis in combination with mass spectrometry, MALDI-MS) revealed a single protein, ubiquitine cross-reactive protein (UBCR), whose level markedly increased due to IFN-alpha treatment. The ubiquitination-like directed degradation processes may thus play a role in the mechanism of IFN-alpha antiproliferative effects. PMID- 14757444 TI - Estimating muscle attachment contours by transforming geometrical bone models. AB - For individualization of a biomechanical model, it is necessary to estimate the muscle attachments of the person to whom it is to be adapted. One of the methods to estimate muscle attachments is to use model transformations to transform a model with known muscle attachments to the bones of a person. We hypothesize that the location and shape of muscle attachment sites correlate with the shape of the bones they are attached to. If this hypothesis holds, it is possible to predict the location of muscle attachments when the shape of the bones is known. To validate this hypothesis, geometric models of three sets of shoulder bones were built. These models consist of 3-D surface models of the scapula, clavicle, and humerus, with the muscle attachment contours connected to them. By means of geometric transformations, the models were transformed, so the muscle attachments of the different data sets could be compared. Using these techniques, 50 per cent of the muscle attachment contours could be predicted with high accuracy. The muscle attachment contours that could not be predicted were all influenced by measurement errors. For 30 per cent of the muscle attachment contours, it was not possible to distinguish the interindividual differences from the inaccuracies of the method used. From this study, we concluded that most muscle attachment contours can be predicted by means of geometric models of the bones. PMID- 14757445 TI - Rotational in vitro compliance measurement of diverse anastomotic configurations: a tool for anastomotic engineering. AB - Anastomotic configurations with a small internal diameter are prone to intimal hyperplasia which can cause occlusion within weeks or months. A link between intimal hyperplasia and inhomogenities of the elastic profile of the anastomosis has been established, making anastomotic engineering directed towards smoothing the compliance profile at the anastomotic site essential. Methods to date restrict the anastomotic compliance measurement to one plane. We present a method by which the anastomotic configurations are rotated, thereby allowing an anastomotic elastic profile assessment in multiple planes. Eight end-to-end anastomoses (ovine common carotid artery) and three end-to-side anastomoses (e PTFE graft to ovine common carotid artery) were prepared and mounted in an artificial circulation system. Anastomotic circumferential compliance (maximal minimal diameter/(maximal-minimal pressure.minimal diameter)) was measured by means of a laser-scan-micrometer and a Statham pressure transducer. By rotating end-to-end anastomoses, the compliance was measured in three, and in end-to-side anastomoses in four different planes. Multiplanar compliance variability in areas remote to both end-to-end and end-to-side anastomoses was approximately 9%. At the suture line the variability was approximately 22% in end-to-end anastomoses and 78% in end-to-side anastomoses. These results show that local factors result in different compliance profiles when utilizing a multiplanar technique, particularly in end-to-side anastomoses. The rotational apparatus is a tool which can be used to more accurately engineer a homogeneously compliant anastomosis, with the ultimate goal of prolonging anastomotic patency. PMID- 14757446 TI - A mathematical model for the laser treatment of heart disease. AB - Transmyocardial laser revascularisation (TMLR) is used to treat patients with severe coronary artery disease. A laser is used to create narrow tunnels within ischemic heart muscle in an attempt to reperfuse the area with oxygenated blood directly from the left-ventricular chamber. It has been hypothesised that initially blood flow through the patent tunnels plays an important role in the efficacy of the treatment (J. Am. College Cordiol. 25(1) (1995) 258) and Waters (J. Fluid Mech. 433 (2001) 193) developed a simple mathematical model to show that this blood flow enhances the quantity of oxygenated blood drawn into the tunnel and the subsequent delivery of oxygen to the tissue. To date, however, the optimum parameter values for this clinical technique have not been determined, e.g. the radius of the laser-drilled tunnels and their relative spacing. We present a mathematical model to determine the distribution of oxygen for a wide range of the governing parameters. Our results indicate that the tunnel radius has a significant effect on the degree of tissue reperfusion and predictions for the optimum tunnel spacing are made. PMID- 14757447 TI - Finite element and experimental analyses of polymer-based dental bridges reinforced by ceramic bars. AB - Dental bridges made of polymer materials reveal only low loading capacity. This paper analyzes possible improvements of the loading capacity of polymer-based dental bridges reinforced by incorporated ceramic bars. Finite element (FE) analyses were performed to study the stress distribution caused by the mastication process in the bridge material. In the experimental part of the study, the fracture load of dental bridges with and without ceramic bar reinforcement was evaluated. Critical stresses occur in the connector area between abutment and pontic of bridges without bar reinforcement. A suitably shaped ceramic bar incorporated into the polymer-based bridge can significantly reduce these critical stresses. The fracture load of the bridge was increased from 515 to 1603N by the bar reinforcement. We conclude from our study that a ceramic bar can significantly improve the loading capacity of a polymer-based dental bridge. The FE and the experimental analyses revealed that the detailed design of the ceramic bar is of decisive importance for the effectiveness of the suggested ceramic reinforcement. PMID- 14757448 TI - Evaluation of a computational model used to predict the patellofemoral contact pressure distribution. AB - One possible cause of patellofemoral pain syndrome is excessive lateral force acting on the patella. Although several treatment methods focus on decreasing the lateral force acting on the patella, the relationship between the lateral force and the patellofemoral contact pressure distribution is unclear. A computational model has been developed to determine how loading variations alter the patellofemoral force and pressure distributions for individual knees. The model allows variation in the quadriceps and patella tendon forces, and calculates the predicted contact pressure distribution using the discrete element analysis technique. To characterize the accuracy of the model, four cadaver knees were flexed on a knee simulator with three initial Q-angles, while recording the force and pressure distributions with a pressure sensor. A model of each knee was created from CT data. Using the external force applied to the knee, the geometry of the knee, and the quadriceps origin as input, the pressure distribution was calculated during flexion. Similar trends were noted for the computational and experimental results. The percentage of the total force applied to the lateral cartilage increased with the Q-angle. The maximum contact pressure increased during flexion. The maximum lateral contact pressure increased with the Q-angle for three knees. For the other knee, increasing the Q-angle decreased the maximum lateral pressure. The maximum medial contact pressure decreased as the Q-angle increased. By characterizing the influence of patellofemoral loading on the force and pressure distributions, the computational model could be used to evaluate treatment methods prescribed for patellofemoral pain. PMID- 14757449 TI - Collagen fibers reduce stresses and stabilize motion of aortic valve leaflets during systole. AB - The effect of collagen fibers on the mechanics and hemodynamics of a trileaflet aortic valve contained in a rigid aortic root is investigated in a numerical analysis of the systolic phase. Collagen fibers are known to reduce stresses in the leaflets during diastole, but their role during systole has not been investigated in detail yet. It is demonstrated that also during systole these fibers substantially reduce stresses in the leaflets and provide smoother opening and closing. Compared to isotropic leaflets, collagen reinforcement reduces the fluttering motion of the leaflets. Due to the exponential stress-strain behavior of collagen, the fibers have little influence on the initial phase of the valve opening, which occurs at low strains, and therefore have little impact on the transvalvular pressure drop. PMID- 14757450 TI - Comparison of shear forces and ligament loading in the healthy and ACL-deficient knee during gait. AB - The purpose of this study was to predict and explain the pattern of shear force and ligament loading in the ACL-deficient knee during walking, and to compare these results to similar calculations for the healthy knee. Musculoskeletal modeling and computer simulation were combined to calculate ligament forces in the ACL-deficient knee during walking. Joint angles, ground-reaction forces, and the corresponding lower-extremity muscle forces obtained from a whole-body dynamic optimization simulation of walking were input into a second three dimensional model of the lower extremity that represented the knee as a six degree-of-freedom spatial joint. Anterior tibial translation (ATT) increased throughout the stance phase of gait when the model ACL was removed. The medial collateral ligament (MCL) was the primary restraint to ATT in the ACL-deficient knee. Peak force in the MCL was three times greater in the ACL-deficient knee than in the ACL-intact knee; however, peak force sustained by the MCL in the ACL deficient knee was limited by the magnitude of the total anterior shear force applied to the tibia. A decrease in anterior tibial shear force was brought about by a decrease in the patellar tendon angle resulting from the increase in ATT. These results suggest that while the MCL acts as the primary restraint to ATT in the ACL-deficient knee, changes in patellar tendon angle reduce total anterior shear force at the knee. PMID- 14757451 TI - Prediction of biomechanical properties of articular cartilage with quantitative magnetic resonance imaging. AB - Quantitative magnetic resonance imaging (MRI) is the most potential non-invasive means for revealing the structure, composition and pathology of articular cartilage. Here we hypothesize that cartilage mechanical properties as determined by the macromolecular framework and their interactions can be accessed by quantitative MRI. To test this, adjacent cartilage disk pairs (n=32) were prepared from bovine proximal humerus and patellofemoral surfaces. For one sample, the tissue Young's modulus, aggregate modulus, dynamic modulus and Poisson's ratio were determined in unconfined compression. The adjacent disk was studied at 9.4T to determine the tissue T(2) relaxation time, sensitive to the integrity of the collagen network, and T(1) relaxation time in the presence of Gd DTPA, a technique developed for the estimation of cartilage proteoglycan (PG) content. Quantitative MRI parameters were able to explain up to 87% of the variations in certain biomechanical parameters. Correlations were further improved when data from the proximal humerus was assessed separately. MRI parameters revealed a topographical variation similar to that of mechanical parameters. Linear regression analysis revealed that Young's modulus of cartilage may be characterized more completely by combining both collagen- and PG-sensitive MRI parameters. The present results suggest that quantitative MRI can provide important information on the mechanical properties of articular cartilage. The results are encouraging with respect to functional imaging of cartilage, although in vivo applicability may be limited by the inferior resolution of clinical MRI instruments. PMID- 14757452 TI - Biological response of the intervertebral disc to dynamic loading. AB - Disc degeneration is a chronic remodeling process that results in alterations of matrix composition and decreased cellularity. This study tested the hypothesis that dynamic mechanical forces are important regulators in vivo of disc cellularity and matrix synthesis. A murine model of dynamic loading was developed that used an external loading device to cyclically compress a single disc in the tail. Loads alternated at a 50% duty cycle between 0MPa and one of two peak stresses (0.9 or 1.3MPa) at one of two frequencies (0.1 or 0.01Hz) for 6h per day for 7 days. An additional group received static compression at 1.3MPa for 3h/day for 7 days. A control group wore the device with no loading. Sections of treated discs were analyzed for morphology, proteoglycan content, apoptosis, cell areal density, and aggrecan and collagen II gene expression. Dynamic loading induced differential effects that depended on frequency and stress. No significant changes to morphology, proteoglycan content or cell death were found after loading at 0.9MPa, 0.1Hz. Loading at lower frequency and/or higher stress increased proteoglycan content, matrix gene expression and cell death. The results have implications in the prevention of intervertebral disc degeneration, suggesting that loading conditions may be optimized to promote maintenance of normal structure and function. PMID- 14757453 TI - Effects of stent stiffness on local haemodynamics with particular reference to wave reflections. AB - The placement of a rigid stent within an elastic vessel produces wave reflection sites at the entrance to and exit from the stent. The net haemodynamic effects of these reflections depend critically on the degree of stiffness of the stent and on its length and position within the diseased vessel, variables that have been found to affect the clinical performance of a stent. Here these effects are examined analytically, using a segmented tube model. The results indicate that the presence of the stent within the larger diseased vessel has the effect of producing higher pressure at the vessel entrance than that at exit. This pressure difference, when superimposed on the underlying pressure distribution within the vessel, has the net effect of actually aiding rather than impeding the flow, but the extent of this depends on the length and position of the stent. A short stent placed near the entrance of the diseased vessel may be favoured clinically for producing the least perturbation in the underlying haemodynamics and thus reducing the chance of restenosis, while a long stent placed near the exit may be favoured for producing a positive pressure difference and thus aiding the flow. PMID- 14757454 TI - Assessment of the functional method of hip joint center location subject to reduced range of hip motion. AB - Motion analysis of the lower extremities usually requires determination of the location of the hip joint center. The results of several recent studies have suggested that kinematic and kinetic variables calculated from motion analysis data are highly sensitive to errors in hip joint center location. "Functional" methods in which the location of the hip joint center is determined from the relative motion of the thigh and pelvis, rather than from the locations of bony landmarks, are promising but may be ineffective when motion is limited. The aims of the present study were to determine whether the accuracy of the functional method is compromised in young and elderly subjects when limitations on hip motion are imposed and to investigate the possibility of locating the hip joint center using data collected during commonly studied motions (walking, sit-to stand, stair ascent, stair descent) rather than using data from an ad hoc trial in which varied hip motions are performed. The results of the study suggested that functional methods would result in worst-case hip joint center location errors of 26mm (comparable to the average errors previously reported for joint center location based on bony landmarks) when available hip motion is substantially limited. Much larger errors ( approximately 70mm worst-case), however, resulted when hip joint centers were located from data collected during commonly performed motions, perhaps because these motions are, for the most part, restricted to the sagittal plane. It appears that the functional method can be successfully implemented when range of motion is limited but still requires collection of a special motion trial in which hip motion in both the sagittal and frontal planes is recorded. PMID- 14757455 TI - Stresses in the local collagen network of articular cartilage: a poroviscoelastic fibril-reinforced finite element study. AB - Osteoarthritis (OA) is a multifactorial disease, resulting in diarthrodial joint wear and eventually destruction. Swelling of cartilage, which is proportional to the amount of collagen damage, is an initial event of cartilage degeneration, so damage to the collagen fibril network is likely to be one of the earliest signs of OA cartilage degeneration. We propose that the local stresses and strains in the collagen fibrils, which cause the damage, cannot be determined dependably without taking the local arcade-like collagen-fibril structure into account. We investigate this using a poroviscoelastic fibril-reinforced FEA model. The constitutive fibril properties were determined by fitting numerical data to experimental results of unconfined compression and indentation tests on samples of bovine patellar articular cartilage. It was demonstrated that with this model the stresses and strains in the collagen fibrils can be calculated. It was also exhibited that fibrils with different orientations at the same location can be loaded differently, depending on the local architecture of the collagen network. To the best of our knowledge, the present model is the first that can account for these features. We conclude that the local stresses and strains in the articular cartilage are highly influenced by the local morphology of the collagen-fibril network. PMID- 14757456 TI - Altered tissue properties induce changes in cancellous bone architecture in aging and diseases. AB - The mechanical properties of cancellous bone depend on its architecture and the tissue properties of the mineralized matrix. The architecture is continuously adapted to external loads. In this paper, it was assumed that changes in tissue properties leading to changes in tissue deformation can induce adaptation of the architecture. We asked whether changes in cancellous bone architecture with aging and in e.g. early osteoarthrosis can be explained from changes in tissue properties. This was investigated using computer models in which the cancellous architecture was adapted to external loads. Bone tissue with deformations below a certain threshold was resorbed, deformations above another threshold induced formation. Deformations between these two boundaries, in the 'lazy zone', did not induce bone adaptation. The effects of changes in bone tissue stiffness on bone mass, global stiffness and architecture were investigated. The bone gain (40-60%) resulting from a 50% decrease in tissue stiffness (simulating diseased tissue) was much larger than the bone loss (2-30%) resulting from a 50% increase in tissue stiffness (simulating highly mineralized, old tissue). The adaptation induced by a decrease in tissue stiffness resulted in an almost constant stiffness in the main load bearing direction, but the transversal stiffness decreased. An increased tissue stiffness resulted in a higher stiffness in the main direction and overcompensation in the transversal directions: the global stiffness could become even smaller than the stiffness of the original model. Concluding, we showed that changes in trabecular bone in aging and diseases can be partly explained from changes in tissue properties. PMID- 14757457 TI - Strain-rate dependence of cartilage stiffness in unconfined compression: the role of fibril reinforcement versus tissue volume change in fluid pressurization. AB - The strain and strain-rate-dependent response of articular cartilage in unconfined compression was studied theoretically. The transient stress and stiffness of cartilage were determined for strain rates ranging from zero to infinity. It is shown, for a given compressive strain, that the axial stress initially increases quickly as a function of strain rate, and then increases progressively more slowly towards the stress corresponding to the instantaneous response. The volume change of the tissue does not give its transient stiffness uniquely, because of the strong strain-rate dependence. The variation of tissue stiffness is primarily determined by the transient stiffness of the radial fibrils. Load sharing between the solid matrix and fluid pressurization also depends on the strain rate. At 15% axial compression, the matrix bears more than 80% of the applied load at a strain rate of 0.005%/s, while the fluid pressurization contributes more than 80% of the load at a strain rate of 0.15%/s. These results show the interplay between fibril reinforcement and fluid pressurization in articular cartilage: the fluid drives fibril stiffening which in turn produces high pore pressure at high strain rates. As a secondary objective of the present work, a fibrillar continuum element was formulated to replace the fibrillar spring element used previously in fibril-reinforced modeling, in order to eliminate the deformation incompatibility between the spring system and the nonfibrillar matrix. The results obtained using the two fibrillar elements were compared with the closed-form solutions for the static and instantaneous responses for the case of large deformation. It was found for unconfined compression that using the spring elements did not generally result in greater numerical errors than using the fibrillar continuum elements. PMID- 14757458 TI - A three-dimensional finite element model of the human anterior cruciate ligament: a computational analysis with experimental validation. AB - In this study, the force and stress distribution within the anteromedial (AM) and posterolateral (PL) bundles of the anterior cruciate ligament (ACL) in response to an anterior tibial load with the knee at full extension was calculated using a validated three-dimensional finite element model (FEM) of a human ACL. The interaction between the AM and PL bundles, as well as the contact and friction caused by the ACL wrapping around the bone during knee motion, were included in the model. The AM and PL bundles of the ACL were simulated as incompressible homogeneous and isotropic hyperelastic materials. The multiple-degrees-of-freedom (DOF) knee kinematics of a cadaveric knee were first obtained using a robotic/universal force-moment sensor testing system. These data were used as the boundary conditions for the FEM of the ACL to calculate the forces in the ACL. The calculated forces were compared to the in situ force in the ACL, determined experimentally, to validate the model. The validated FEM was then used to calculate the force and stress distribution within the ACL under an anterior tibial load at full extension. The AM and PL bundles shared the force, and the stress distribution was non-uniform within both bundles with the highest stress localized near the femoral insertion site. The contact and friction caused by the ACL wrapping around the bone during knee motion played the role of transferring the force from the ACL to the bone, and had a direct effect on the force and stress distribution of the ACL. This validated model will enable the analysis of force and stress distribution in the ACL in response to more complex loading conditions and has the potential to help design improved surgical procedures following ACL injuries. PMID- 14757460 TI - A method to estimate the elastic properties of the extracellular matrix of articular cartilage. AB - In this work we propose a method to estimate the elastic properties of the extracellular matrix of articular cartilage, once the elastic properties of the chondrocytes and the whole tissue are known. The influence of the elastic properties of the tissue and the cell concentration on the estimated elastic properties of the matrix are investigated. PMID- 14757459 TI - The correspondence between equilibrium biphasic and triphasic material properties in mixture models of articular cartilage. AB - Mixture models have been successfully used to describe the response of articular cartilage to various loading conditions. Mow et al. (J. Biomech. Eng. 102 (1980) 73) formulated a biphasic mixture model of articular cartilage where the collagen proteoglycan matrix is modeled as an intrinsically incompressible porous permeable solid matrix, and the interstitial fluid is modeled as an incompressible fluid. Lai et al. (J. Biomech. Eng. 113 (1991) 245) proposed a triphasic model of articular cartilage as an extension of their biphasic theory, where negatively charged proteoglycans are modeled to be fixed to the solid matrix, and monovalent ions in the interstitial fluid are modeled as additional fluid phases. Since both models co-exist in the cartilage literature, it is useful to show how the measured properties of articular cartilage (the confined and unconfined compressive and tensile moduli, the compressive and tensile Poisson's ratios, and the shear modulus) relate to both theories. In this study, closed-form expressions are presented that relate biphasic and triphasic material properties in tension, compression and shear. These expressions are then compared to experimental findings in the literature to provide greater insight into the measured properties of articular cartilage as a function of bathing solutions salt concentrations and proteoglycan fixed-charge density. PMID- 14757461 TI - Prediction of lubrication regimes in wrist implants with spherical bearing surfaces. AB - The wrist joint is frequently affected by rheumatoid arthritis, resulting in wrist pain, deformity and ultimately loss of function. Artificial wrist implants have been introduced to treat the rheumatoid wrist, to attempt to alleviate pain and restore some function to the joint. The aim of this study was to predict the likely lubrication regimes that occur in wrist implants with spherical bearing surfaces. The implant was modelled as an equivalent ball-on-plane. Elastohydrodynamic lubrication theory was used to determine the minimum film thickness for the implant under different load, entraining velocity, lubricant viscosity, size of implant and material combinations. The results show that the highest film thickness is found in large implants, with high viscosity, high entraining velocity and low load. Hard-on-soft material combinations will operate with a boundary lubrication regime. Material combinations involving ceramic bearing surfaces have the potential to operate with a mixed lubrication regime. PMID- 14757462 TI - Estimation of the centre of rotation: a methodological contribution. AB - The location of the centre of rotation of human joints that can be modelled as a spherical hinge can be estimated using kinematics information about the two adjacent bony segments involved recorded while the subject makes them move one relative to the other (functional method). In order to solve the relevant analytical problem, several algorithms have been proposed. Most recently, two methods, one based on a spherical best-fit approach and another based on the Reuleaux construction, have been presented as being different and submitted to comparative evaluation. This paper modifies the second method taking all information in the data set into account and shows that, having done this, the two methods coincide analytically. PMID- 14757463 TI - Outflow distribution at the distal anastomosis of infrainguinal bypass grafts. AB - Outflow distribution at the distal anastomosis of infrainguinal bypass grafts remains unquantified in vivo, but is likely to influence flow patterns and haemodynamics, thereby impacting upon graft patency. This study measured the ratio of distal to proximal outflow in 30 patients undergoing infrainguinal bypass for lower limb ischaemia, using a flow probe and a transit-time ultrasonic flow meter. The mean outflow distribution was approximately 75% distal to 25% proximal, with above knee anastomoses having a greater proportion of distal flow (84%) compared to below knee grafts (73%). These in vivo flow characteristics differ significantly from those used in theoretical models studying flow phenomena (50:50 and/or 100:0), and should be incorporated into future research. PMID- 14757464 TI - Determination of orthotropic bone elastic constants using FEA and modal analysis by Taylor WR et al. [J Biomech. (2002) Vol. 35, pp. 767-773]. PMID- 14757465 TI - Thermal factors affecting stability and durability of cemented metal prostheses. PMID- 14757466 TI - Color Doppler ultrasound evaluation of testicular blood flow in stallions. AB - The objectives of this study were to evaluate the potential use of color Doppler ultrasound to characterize blood flow to the stallion testis, and to establish reference values for Doppler measures of blood flow in the testicular artery of the stallion. Both testes from each of 52 horses were examined using a pulsed wave color Doppler ultrasound with a sector array 5/7.5 MHz transducer with a 1mm gate setting. Peak systolic velocity (PSV), end diastolic velocity (EDV), resistive index (RI), and pulsatility index (PI) of the testicular artery were measured in each of two locations, the convoluted aspect (spermatic cord) and the marginal aspect of the artery (on the epididymal edge of testis). We found that: (1) all measures were obtainable; (2) except for EDV, the majority of the measures were higher at the cord location than at the marginal aspect of the artery (P < 0.05); and (3) measures for left and right testes were similar (P > 0.10). Resulting measures from 41 of these stallions (82 testes) that appeared free of testicular pathology provide useful reference values for clinical evaluation. Evaluation of 11 cases with testicular pathology suggested further investigation of possible effects of these various conditions on testicular blood flow and testicular function. PMID- 14757467 TI - Live cubs born after transfer of OPS vitrified-warmed embryos in the farmed European polecat (Mustela putorius). AB - The Open Pulled Straw (OPS) method of vitrification has been used successfully for cryopreserving embryos of most domestic animal species. However, there is no report of a successful delivery of offspring after transfer of vitrified embryos in carnivores, even though vitrification has been a successful freezing method for species like swine whose embryos are known to be susceptible to chilling injury. Morulae and blastocysts of farmed European polecat (Mustela putorius) were vitrified and warmed before in vitro culture in modified synthetic oviductal fluid (SOF) for a period from a few hours up to 3 days before being transferred to recipients. Survival rate after vitrification, warming and in vitro culture was 51% (50/98). A total of 50 embryos were transferred surgically into the uteri of four anesthetized recipients. Two recipients delivered a total of eight offspring (2 and 6 each) for an overall survival rate of 16% (eight live cubs/50 transferred embryos). According to our knowledge, these offspring are the first carnivores produced by transfer of in vivo embryos after vitrification by OPS. Based on the present results, we suggest that OPS vitrification can be used as an alternative cryopreservation method for mustelid embryos with pup results comparable to conventional slow freezing. PMID- 14757468 TI - Observations on the fertilization and development of preimplantation bovine embryos in vitro in the presence of Tritrichomonas foetus. AB - Tritrichomonas foetus, a world-wide distributed parasitic protozoan is a cause of infertility and abortion. There is no documented information on the susceptibility of bovine embryos to the parasite. To determine the effect of T. foetus on fertilization and embryonic development of preimplantation bovine embryos, we added approximately 10(4)/ml or 10(6)/ml T. foetus (Belfast strain) to sperm cells and oocytes prior to in vitro fertilization (IVF) or to presumptive zygotes 24 h post-fertilization. Light and scanning electron microscopy (SEM) revealed that exposure of oocytes or embryos at any stage of development to T. foetus caused rapid adhesion of the trichomonads to the embryonic intact zona pellucida (ZP) and to trophoblastic cells of hatched blastocysts. Treatment of contaminated embryos with 0.25% trypsin for 3 min did not render them free from T. foetus. Motile parasites were not observed after 18 h incubation in IVF medium, or after 72 h in synthetic oviductal fluid (SOF) embryo culture medium. The percentages of cleaved zygotes, blastocysts and hatched embryos resulting from culture of experimental and uninfected control groups of embryos were not different (P > 0.05). Tritrichomonas foetus was not detected in embryonic cells of ZP-intact or hatched embryos when examined by transmission electron microscopy (TEM). In conclusion, T. foetus has no detrimental effect on the fertilization and development of IVF embryos and the potential risk of transmission of trichomonosis is unlikely, due to the limited survival of the parasite in IVF culture conditions. PMID- 14757469 TI - Effect of cryoprotectants and their concentration on in vitro development of vitrified-warmed immature oocytes in buffalo (Bubalus bubalis). AB - Experiments were conducted to study the effect of cryoprotectants, dimethyl sulfoxide (DMSO), ethylene glycol (EG), 1,2-propanediol (PROH), and glycerol at different concentrations (3.5, 4, 5, 6, and 7 M each with 0.5 M sucrose and 0.4% BSA in DPBS) on survival, in vitro maturation, in vitro fertilization, and post fertilization development of vitrified-thawed immature buffalo oocytes. The COCs were harvested from the ovaries by aspirating the visible follicles. The recovery of post-thaw morphologically normal oocytes was lower in 3.5 and 4 M DMSO, EG, and PROH compared to 5, 6, and 7 M. In all the concentrations of glycerol, an overall lower numbers of oocytes recovered were normal compared to other cryoprotectants. Less number of oocytes reached metaphase-II (M-II) stage from the oocytes cryopreserved in any of the concentrations of DMSO, EG, PROH, and glycerol compared to fresh oocytes. Among the vitrified groups, highest maturation was obtained in 7 M solutions of all the cryoprotectants. The cleavage rates of oocytes vitrified in different concentrations of DMSO, EG, PROH, and glycerol were lower than that of the fresh oocytes. The cleavage rates were higher in oocytes cryopreserved in 6 and 7 M DMSO, EG, PROH, and glycerol compared with oocytes cryopreserved in other concentrations. However, the percentage of morula and blastocyst formation from the cleaved embryos did not vary in fresh oocytes and vitrified oocytes. In conclusion, this report describes the first successful production of buffalo blastocysts from immature oocytes cryopreserved by vitrification. PMID- 14757470 TI - Transfer of vitrified blastocysts from one or two superovulated Large White Hyperprolific donors to Meishan recipients: reproductive parameters at Day 30 of pregnancy. AB - The present study was designed to determine the effect of pooling embryos from two donors on the reproductive success of transfer of vitrified/warmed porcine blastocysts. Intact blastocysts were collected from superovulated Large White Hyperprolific gilts (n = 24) on Days 5-5.5 after artificial insemination. Embryos were recovered by flushing the uterine horns, and unhatched blastocysts were selected. Vitrification and warming were performed as described by Berthelot et al. [Cryobiology 41(2000) 116]. To evaluate in vitro development, 37 vitrified/warmed blastocysts were cultured, non-vitrified embryos (n = 48) were used as controls. Embryo transfers were conducted in asynchronous (-24 h) Meishan gilts (n = 20). Twenty vitrified/warmed blastocysts were surgically transferred into one uterine horn. Ten recipients received embryos from one donor (Group 1) and the other 10 transfers were performed with mixed embryos from two donors (Group 2). Pregnancy was assessed ultrasonographically at Day 25 after estrus and recipients were slaughtered at Day 30 after transfer. In vitro survival rate of the vitrified/warmed blastocysts was lower (P < 0.01) than that from control embryos (73.0% versus 93.7%). The pregnancy rate for Group 1 (70%) was not different (P > 0.05) than that from Group 2 (90%). No significant differences were detected between Groups 1 and 2 for in vivo embryo development (number fetuses/transferred embryos in pregnant recipients) or in vivo embryo survival (number viable fetuses/transferred embryos in pregnant recipients). However, the in vivo efficiency (number viable fetuses/total transferred embryos) was higher (P < 0.05) when transfers were performed with embryos from two donors (19.5% versus 30.5%). These results indicate that pooling embryos from two donors increases the in vivo efficiency after transfer of vitrified/warmed porcine blastocysts. PMID- 14757471 TI - Retained placenta in Friesian mares: incidence, and potential risk factors with special emphasis on gestational length. AB - During the foaling seasons of 1999 and 2000, the incidence of retained placenta in 495 normal parturitions of 436 Friesian brood mares was studied. Retained placenta was defined as a failure to expel all fetal membranes within 3 h of the delivery of the foal. Furthermore, the sex of the foal, month of breeding, sire and dam's sire, age of the mare, and time of day of foaling, were studied as factors that might be associated with retained placenta in Friesian mares after normal foalings, and with gestational length. The analysis was carried out using marginal logistic regression, and mixed linear regression, respectively. The incidence of retained placenta was 54%. Mean length of gestation was 331.6 days. Colts were carried 1.5 days longer than fillies. Mares bred in July-September had a 4-day shorter gestation period (329 days) than mares bred earlier in the year. There was a mare, sire, and dam's sire effect on gestational length, and a mare effect on the occurrence of retained placenta. Mares foaling at 4 and >17 years of age, tended to have a lower incidence of retained placenta than mares foaling at 5-17 years of age. No association was found between the occurrence of retained placenta, and gestational length, sex of the foal, month of breeding, dam's sire, and time of day of foaling. It was concluded that the observed high incidence of retained placenta indicates that the Friesian breed of horses has a higher risk for retained placenta than other breeds of horses. PMID- 14757472 TI - Occurrence of flehmen in male buffaloes (Bubalus bubalis) with special reference to estrus. AB - Pheromonal cues present in the urine and vaginal mucus during the estrus period elicit courtship behaviour in mammals including the characteristic flehmen behaviour. In the present study, the flehmen behaviour was assessed in male buffaloes by exposing them to heifers for three consecutive estrous cycles. Behavioural observations revealed that the bulls sniffed the female's external genitalia, responded to the chemical signal(s) and exhibited the flehmen behaviour similar to those reported in other ungulates. The flehmen behaviour was recorded daily during 15-min contact with females for three consecutive estrous cycles. Out of 180 observations during five days estrus periods (-3 to +1 days) which included high frequency of flehmen behaviour and out of 637 observations with or without flehmen behaviour during diestrus periods, the bull displayed 365 and 441 flehmen behaviour, respectively. Average numbers of all (2.03 +/- 0.66) and repeated flehmen (1.05 +/- 0.64) behaviour during estrus periods were significantly higher than those of diestrus periods of all (0.69 +/- 0.25) and repeated flehmen (0.11 +/- 0.10). The statistical significance was higher (P < 0.001) in repeated flehmen behaviour of estrus as compared to that in diestrus. In case of all flehmen behaviour, the statistical significance was higher (P < 0.01) in estrus when compared to that in diestrus. The occurrence of flehmen behaviour in male buffaloes clearly indicates that the specific pheromonal compound(s) present in the urine/vaginal mucus during estrus significantly influence the flehmen behaviour. PMID- 14757473 TI - Fetometry and fetal heart rates between Day 35 and 108 in bovine pregnancies resulting from transfer of either MOET, IVP-co-culture or IVP-SOF embryos. AB - The Large Offspring Syndrome has frequently been reported for in vitro produced calves. The objective of this study was to determine whether any differences in body dimensions (biparietal diameter of the cranium (BPD), cross-section of the abdomen at the insertion of the umbilical cord (CAU)) and heart rate (FHR) can be detected during the first 108 days of gestation between bovine foetuses derived from different methods of embryo production. Three groups of pregnancies with calvings at term resulted from non-surgical transfers of three types of embryos: recipients carrying an embryo obtained by standard MOET procedures (n = 25); recipients carrying an embryo produced in vitro from OPU-derived oocytes, using co-culture-medium (n = 14) or SOF-medium (n = 22). Transrectal ultrasonographic examinations were performed weekly. Ultrasound images were recorded and during off-line analysis FHR, BPD and CAU were determined. For each foetus a curve was fitted and the estimates on fixed time intervals were used as dependent variables in an analysis of variance to detect differences between the three pregnancy groups. Neither gestation length nor birth weight differed significantly between the three pregnancy groups, nor could any differences with respect to BPD, CAU or FHR be detected between Days 35 and 108 of gestation. It is concluded that no differences exist between the early development of bovine foetuses, derived from MOET, IVP-co-culture or IVP-SOF embryos, and resulting in calves with normal birth weights. PMID- 14757474 TI - Body weight of mares and foals, estrous cycles and plasma glucose concentration in lactating and non-lactating Lipizzaner mares. AB - This study summarizes weight development, plasma glucose concentrations and reproductive parameters in lactating (n = 46) and non-lactating Lipizzaner mares (n = 11) throughout the breeding season. It was the aim of the study to analyse if an energy deficit with possible effects on reproductive functions occurs at any time during the first 4 months of gestation. Mean gestation length was 334.3 +/- 7.3 days. Gestation of foals born in May/June was shorter (P < 0.01) than for foals born in March/April. Out of the 46 lactating mares, 44 ovulated between Days 8 and 18 postpartum and two mares ovulated on days 30 and 145, respectively. Pregnant mares were significantly (P < 0.001) heavier (600.1 +/- 5.3 kg) than non pregnant mares (521.8 +/- 10.0 kg) at the beginning of the study. Birth resulted in weight reduction of 64.8 +/- 2.4 kg. During the first 2 weeks postpartum mares lost on average 3.0 +/- 1.8 kg and in the following 2 weeks gained 3.6 +/- 1.4 kg of weight. Thereafter, weight increased slightly but continuously (P < 0.01). At no time after foaling, weight differed significantly between groups. Weight of the foals three days after birth varied between 29 and 67 kg (53.7 +/- 1.1 kg). Average daily weight gain of foals was relatively constant throughout the study period (1.15 +/- 0.17 kg). Although lactation at no time was associated with a major weight loss, it had clear effects on energy metabolism as shown by constantly lower plasma glucose concentrations in lactating mares. Glucose concentrations decreased after foaling and were significantly lower in lactating mares from Weeks 3 to 16 after foaling than at corresponding times in non lactating mares (P < 0.01). However, glucose concentrations were still within the physiological range. Mares seem to be able to compensate energy losses during lactation mainly by increasing feed intake and not by mobilisation of body fat. PMID- 14757475 TI - Bull semen in vitro fertility after cryopreservation using egg yolk LDL: a comparison with Optidyl, a commercial egg yolk extender. AB - Low-density lipoproteins (LDL) have been previously isolated and identified as the cryoprotective fraction of yolk. The effect of LDL on sperm motility after freezing-thawing has been reported, but no study has been made to assess the effect of LDL on bull semen fertility. The aim of this study was to evaluate the fertility of bull semen cryopreserved in the presence of LDL. Motility of semen cryopreserved in LDL was analyzed and compared to semen cryopreserved with Optidyl, a commercial extender containing egg yolk. To evaluate the fertilizing ability of semen, we used in vitro fertilization test, whereas acrosome and plasma membrane integrity were also evaluated. The percentage of motile spermatozoa was two fold higher after freezing in LDL than in Optidyl 54.4% versus 30.2% (P < 0.05). The cleavage rate was significantly higher after fertilization with semen frozen in LDL than with Optidyl 63.0% versus 54.8% (P < 0.05). No significant difference was observed on the blastocyst rate after in vitro culture. Integrity of the acrosome and the plasma membrane were maintained in both extenders. In conclusion, LDL preserve bull semen quality and fertilizing ability, allowing also better semen motility, after the freeze-thaw process. PMID- 14757476 TI - Effect of different stages of the follicular wave on in vitro developmental competence of bovine oocytes. AB - This study aimed to investigate the developmental competence of ovum pick-up collected oocytes on three stages of the follicular wave: Days 2, 5 and 8. A group of 11 cows was used in successive cycles to perform ovum pick-up on either Day 2, 5 or 8 of an induced follicular wave (three sessions per stage). Follicular waves were initiated by puncturing the dominant follicle and all other follicles sized > or = 5 mm at Days 5-7 of the cycle. The plasma progesterone concentrations did not differ between the days of ovum pick-up: 4.0 +/- 1.8, 5.1 +/- 1.6 and 5.2 +/- 1.7 ng/ml for Days 2, 5 and 8, respectively. The proportion of oocytes with three or more layers of non-expanded cumulus cells was higher for Day 5 than Day 8, while Days 2 and 5 did not significantly differ from each other (85, 96 and 68% of 113, 60 and 101 oocytes for Days 2, 5 and 8, respectively). The proportion of oocytes competent to develop a blastocyst in an in vitro production system was higher for Days 2 and 5 than for Day 8: 27, 29 and 15% for the oocytes with fair to good cumulus investment and 23, 27 and 11%, respectively, when all oocytes were taken in account. This indicates that the dominant follicle reduces the developmental competence of oocytes from subordinate follicles at a relatively late stage of dominance. This finding has practical consequences for the handling of cows that undergo ovum pick-up only once or very irregularly. The embryo yield can then be improved by performing the ovum pick-up at Days 2-5 of the cycle or 2-5 days after ablation of the large follicles. PMID- 14757477 TI - Evaluation of viability and apoptosis in horse embryos stored under different conditions at 5 degrees C. AB - The aim of this study was to evaluate the viability (percentage of dead cells) and the incidence of DNA fragmentation of horse embryos after storage in three different media at 5 degrees C for 6 and 24 h. Forty embryos were stored in Emcare Holding Solution for 6 and 24 h, in Hams'F10 or Vigro Holding Plus for 24 h at 5 degrees C (n = 9-10 per group) and 10 embryos were evaluated immediately after collection. First, embryos were stained, immediately after collection or following storage, to detect dead cells (DAPI) and, subsequently, DAPI-stained embryos were fixed and stained to detect DNA fragmentation (TUNEL). Finally, all the fixed embryos were re-stained with DAPI to determine the total number of cells. The percentage of cells stained with both TUNEL and DAPI or TUNEL-only or DAPI-only were determined. The percent of dead cells (DAPI-labelled) per embryo increased with duration of storage, but no differences were detected between the storage media. The percentage of early apoptotic cells (TUNEL+/DAPI-) in fresh and stored embryo for 6 h or 24 h did not differ significantly (P > 0.05). There was a significant correlation between the percentage of cells labelled by TUNEL and DAPI (R = 0.87) (P < 0.001). These results suggest that cooled storage increases cell death but this does not appear to occur by induction of apoptosis and that DAPI staining proves to be a quick and reliable method for assessing embryo viability. PMID- 14757478 TI - An efficient method for blocking the 1st mitotic cleavage of fish zygote using combined thermal treatment, exemplified by mud loach (Misgurnus mizolepis). AB - Blocking the first mitotic cleavage of the zygote is a key tool for chromosome set manipulations in fish. We developed an improved method for inducing tetraploidy by blocking the mitosis with a combination of heat shock at 40.5 degrees C for 1, 2, or 3 min followed by cold shock at 1.5 degrees C for 30, 45, or 60 min. When applied during the first cleavage metaphase of mud loach (Misgurnus mizolepis) zygotes, the optimal combination was heat for 2 min followed by cold for 45 min. At 1 month, the frequency of 4N survivors and the yield from total eggs fertilized was 55.7 and 14.4%, respectively, compared to heat shock alone with 20.0% efficiency and 3.6% yield. The effectiveness of the procedure was confirmed by diploid mitotic gynogenesis using transgenic markers. The overall yield of homozygous diploids, 34.0%, was better than that for single heat shock, 17.3%. The tetraploids and homozygous diploids had higher early mortality than normal diploid controls. However, at 1 month, the viability of the tetraploids was the same as normal diploids. For gynogenetic diploids, the survival was similar to normal diploids after 3 months. The high efficiency of this new protocol extends the opportunity to study polyploidy in basic and applied research. PMID- 14757479 TI - Number of oocytes obtained from cows by OPU in early, but not late lactation increased with plasma insulin and estradiol concentrations and expression of mRNA of the FSH receptor in granulosa cells. AB - The effects of lactation stage and hormonal profile on the quality and quantity of oocytes and the gonadotrophic sensitivity of granulosa cells (GC) from small antral follicles obtained by sequential aspirations from ovaries of high producing dairy cows were examined. Cows in late lactation (263(+/- 60) days postpartum) and 98(+/- 16) days pregnant in positive energy balance (EB) showed no significant changes in plasma concentrations of estradiol, progesterone, insulin, IGF1 or expression of mRNA of the FSH receptor in GC from small antral follicles during the 49 days experimental period. There were no changes in the number and quality of oocytes obtained from each aspiration. In cows in early lactation (72.8 +/- 6 days postpartum), plasma insulin concentrations increased and were positively correlated with plasma estradiol concentration. Due to the sequential aspirations progesterone blood concentrations were low in early lactation cows. Expression of mRNA of the FSH receptor increased in GC from small antral follicles of early lactation cows together with the number of oocytes obtained with aspiration sessions. No differences were found in morphological quality or function between oocytes obtained from small antral follicles from cows in early or late lactation. In early, but not late lactation, the number of oocytes was correlated with both insulin and E2 plasma concentrations. Improved EB and sensitivity of GC to FSH may be involved in oocyte recruitment in early lactation. PMID- 14757480 TI - Steroid receptors in canine endometrial cells can be regulated by estrogen and progesterone under in vitro conditions. AB - The expression of estrogen (ER) and progesterone receptors (PR) in the endometrium is regulated by steroid hormones. An increase in plasma estrogen leads to upregulation of the number of both steroid receptors, whereas a decrease in both receptors population is due to high concentration of plasma progesterone. To study the exact effect of different concentrations of beta-estradiol and progesterone on canine epithelial and stromal endometrial cells an in vitro model from dog uterus was developed and kept for 20 days. Material was obtained from healthy dogs, undergoing ovariohysterectomy. Endometrial epithelial and stromal cells were gained after collagenase treatment, followed by filtration steps. Electron microscopy and immunolabeling were used to study cell morphology and differentiation. Immunocytochemistry was used to determine proliferation rate (Ki 67), ER and PR status on Days 3, 8, 10, 13, and 20. Mitotic activity of both cells was stimulated with different concentrations of steroids and revealed high values until cells reached confluency. ER and PR expression in confluent layer from epithelial and stromal cells was upregulated with beta-estradiol. In addition progesterone significant downregulated both receptors population in stromal cells, whereas the reduction was less pronounced in epithelial cells. Results showed that our in vitro system is a useful tool to study the influence of beta-estradiol and progesterone on cell proliferation rate, ER and PR expression. The primary cell culture model helps to avoid experiments on living animals. PMID- 14757481 TI - Effect of GnRH antagonists treatment on gonadotrophin secretion, follicular development and inhibin A secretion in goats. AB - The aim of this study was to determine, for goats, the effects of daily doses of GnRH antagonist on ovarian endocrine and follicular function. Ten does were given 45 mg FGA intravaginal sponges and then five were treated with daily injections of 0.5mg of the GnRH antagonist Teverelix for 11 days from 2 days after the day of sponge insertion, while five does acted as controls. Pituitary activity was monitored by measuring plasma FSH and LH daily from 2 days before the first GnRH injection to Day 12. Follicular activity was determined by ultrasonographic monitoring and by assessing plasma inhibin A levels during the same period. In treated does, the FSH levels decreased linearly (0.8 +/- 0.1 ng/ml to 0.5 +/- 0.1 ng/ml, P < 0.01) and remained lower than the mean concentration in control goats (0.8 +/- 0.1 ng/ml, P < 0.005). LH levels were also lower during the period of antagonist treatment (0.6 +/- 0.2 ng/ml versus 0.4 +/- 0.1 ng/ml, P < 0.0005). During GnRH antagonist treatment, there was a significant decrease in the number of large follicles (> or = 6 mm) from Day 3 of treatment (1.2 +/- 0.6, P < 0.0001), with no large follicles from Day 9. The number of medium follicles (4-5 mm in size) also decrease during the period of treatment (4.2 +/- 0.7 to 1.0 +/- 0.6, P < 0.0001), leading to a significant decrease in inhibin A levels when compared to the control (143.7 +/- 31.3 pg/ml versus 65.2 +/- 19.1 pg/ml, P < 0.00005). In contrast, the number of small follicles (2-3 mm) increased in treated goats from Day 4 of treatment (9.6 +/- 2.9 to 20.2 +/- 6.3, P < 0.005). Such data indicate that GnRH antagonist reduced plasma levels of FSH and LH with suppression of the growth of large dominant ovarian follicles and a two-fold increase in number of smaller follicles. The results confirm that GnRH antagonist treatment can be used in goats to control gonadotrophin secretion and ovarian follicle growth in superovulatory regimes. PMID- 14757482 TI - Studies of antimicrobial activity of N-alkyl and N-acyl 2-(4-thiazolyl)-1H benzimidazoles. AB - Various N-alkyl and N-acyl derivatives of 2-(4-thiazolyl)-1H-benzimidazole, an anthelmintic and systemic fungicide, were synthesized by polymer-supported reactions and screened for their antifungal and antibacterial potency to establish structure-activity relationships. PMID- 14757483 TI - A study on the effect of drying techniques on the mechanical properties of pellets and compacted pellets. AB - Microcrystalline cellulose (MCC) pellets produced by a standard extrusions/spheronisation process with a 40% ethanol/water mixture as the fluid component, were dried by four different techniques, namely: freeze-drying, fluid bed drying, hot air oven drying and desiccation with silica-gel to less than 5% (w/w) water content. A 1.0-1.18mm size fraction of the dried pellets were characterised structurally and mechanically in terms of, shape, density/porosity (open and closed), pore volume/pore volume distribution, surface area, surface tensile strength, shear strength, deformability, linear strain and elastic modulus. An amount of 600, 700 and 750mg of the same size fraction of each pellet batch were compacted to the same tablet thickness and the tensile strength and volumetric elastic recovery of the resulted compacts were determined. Analysis of variance was used to assess the significance of the drying process on the property of the pellets and their compacts. The drying process did not influence the shape of the pellets, but all the other properties were affected to some extent. Pellets dried by freeze-drying were more porous, with most of the pores open to the atmosphere and had a higher surface area than pellets dried by the other methods. Pellets dried by desiccation contained the highest proportion of closed pores. The decrease in tensile strength of the pellets, which occurred with the increase in porosity could presumably be due to ease of crack initiation and propagation between the MCC fibres. The weaker pellets broke instantly before they were subjected to appreciable strain. The porous pellets needed a higher compressing pressure and work of compaction to produce tablets of the same mass and dimensions. This reflected their compressibility, i.e. relative decrease in volume of the pellet bed during compression. The strength and volumetric elastic recovery of the compacts increased with the increase of their porosity. The drying techniques, which produced porous, deformable and weak pellets, produced stronger tablets. The value of the volumetric elastic recovery of the compacts was also observed to increase with the value of compaction pressure. PMID- 14757484 TI - Immunogenicity of meningococcal PorA formulations encapsulated in biodegradable microspheres. AB - The purpose of our study was to investigate the possibility to microencapsulate liposomes and meningococcal outer membrane vesicles (OMV), both containing neisserial pore protein A (PorA), in biodegradable dextran- and mannan-based microspheres and to study the immunogenicity of the encapsulated PorA formulations. PorA-liposomes and OMV were encapsulated in dextran- or mannan based microspheres by using an aqueous two-phase system consisting of a polyethylene glycol solution and a methacrylated dextran or mannan solution. The formulations were characterized for size distribution, PorA structure and antigen recovery after release. Calcein-containing model liposomes were used to establish the encapsulation efficiency and release profiles from both types of microspheres. The immunogenicity of the PorA-containing formulations was determined in mice after subcutaneous immunization. Liposomes were encapsulated in dextran and mannan microspheres with a high efficiency (70-90%). Calcein liposomes, after a 5-day lag period, exhibited apparent zero-order release kinetics from both types of microspheres between Days 5 and 10 of incubation in vitro. The total release was 80 and 100% from mannan and dextran microspheres, respectively. The trimeric PorA conformation was preserved in the released liposomes and OMV and the antigen was partly recovered. The immunogenicity of PorA-liposomes and OMV encapsulated in dextran or mannan microspheres was preserved. In conclusion, PorA-liposomes and OMV could be encapsulated in dextran and mannan-based microspheres with high efficiency. The immunogenicity of encapsulated antigen was preserved. PMID- 14757485 TI - The application of non-contact laser profilometry to the determination of permanent structural changes induced by compaction of pellets. I. Pellets of different composition. AB - Pellets of different mechanical properties were produced by extrusion and spheronisation process based on various composition (microcrystalline cellulose (MCC), lactose, glyceryl monostearate (GMS), water, ethanol and glycerol). Six hundred milligrams of these pellets were compacted at two different pressures (86.7 and 130MPa) to form flat-faced tablets and stored for 48h in ambient temperature and humidity after which their permanent structural change (plastic deformation) was investigated in terms of surface roughness parameters of both faces of the tablets, using a non-contact laser profilometer. The results were compared with the deformability values obtained from the force/displacement curve as well as the phase angle values obtained from the dynamic scan of a dynamic mechanical analyser. The increase in deformability of the pellets with the increase in porosity, increase in the contents of GMS, lactose or ethanol in the formulations and increase in compaction pressure was illustrated by the reduction of the surface roughness parameters. Analysis of variance identified the significant difference in the mean rugosity values of the tablets from the various formulations, tablet faces and compaction pressures. The deformability values obtained were reasonably comparable to those plasticity values obtained in terms of phase angle from the dynamic scan of the DMA and the force/displacement curves. The laser profilometry technique in conjunction with scanning electro microscopy (SEM) demonstrated the low and broad based wavelike structure of the pellets after compaction rather than spiky sharp protrusions. This confirms the capability of determining the plastic deformability of the pellets from the surface roughness parameters obtained from non-contact laser profilometry. PMID- 14757486 TI - Feasibility study for the rapid determination of the amylose content in starch by near-infrared spectroscopy. AB - Near-infrared (NIR) spectroscopy was used to determine the amylose content in six different starches, whose declared amylose contents ranged from 2 to 95% m/m. The amylose content of starches can vary considerably between batches depending on growth conditions and time of harvesting. An NIR calibration model was developed for amylose using simple laboratory produced mixtures of amylose and amylopectin in different ratios. The spectral region at 1700-1800nm showed a good correlation to the amylose content of these mixtures. A simple absorbance ratio calibration model using standard normal variate and first derivative pre-treated spectra gave a root mean standard error of prediction of 1.2% m/m. Application to real samples gave amylose contents in reasonable agreement with the average values stated by the supplier. NIR spectroscopy provides a rapid and non-destructive method for the quantitative determination and standardisation of amylose in starch and could make a suitable alternative to traditional techniques, such as complex formation of starch with iodine or n-butanol. PMID- 14757487 TI - Biocompatibility and integrin-mediated adhesion of human osteoblasts to poly(DL lactide-co-glycolide) copolymers. AB - The biocompatibility of polylactic acid (PLA) and polyglycolic acid (PGA) copolymers, employed in manufacturing bone-graft substitutes, is affected by their chemical composition, molecular weight and cell environment, and by the methods of polymerization and processing. Their in vitro bioactivity on human osteoblasts has been investigated very little. We first evaluated the behavior of primary human osteoblasts cultured in close contact with 75:25 and 50:50 PLA-PGA copolymers for 14 days adopting a cell culture system that allowed us to evaluate the influence of direct contact, and of factors released from polymers. The copolymers had no negative influence on cell morphology, cell viability and proliferation. Alkaline phosphatase (ALP) activity and osteocalcin production were also not affected. The initial adhesion of osteoblasts on implant surfaces requires the contribution of integrins, acting as a primary mechanism regulating cell-extracellular matrix (ECM) interactions. We observed that adhesion of osteoblasts to PLA-PGA copolymers, 2h after plating, was reduced by approximately 70% by antibodies capable to block integrin beta(1) and alpha(5)beta(1) complex and only by approximately 30% by an anti-integrin alpha(v) antibody. Therefore, beta(1) integrins may represent a predominant adhesion receptor subfamily utilized by osteoblasts to adhere to PLA-PGA copolymers. These materials do not show any negative influence on cell proliferation and differentiation. PMID- 14757488 TI - Cost-minimization analysis of treatment of gastroesophageal reflux disease. Implications of varying holding time on conclusions. AB - Gastroesophageal reflux disease (GERD), is a common disorder. The most effective medical treatment for GERD is a proton pump inhibitor (PPI). The aim of this study was to specify the most inexpensive PPI therapy for GERD, and to examine the implications of varying outcome measure, holding time, on the conclusions about the cost-effectiveness of the treatments. Proton pump inhibitors that have holding time of intragastric pH>4 for at least 11h in 24h period (esomeprazole, lansoprazole, omeprazole and rabeprazole), were included. In this cost minimization analysis (CMA), data on holding times were gathered from scientific publications listed in MEDLINE, prices of proton pump inhibitors from the Finnish database of drug prices and the treatment dosages were taken from the official guide of drug therapies in Finland. A decision tree was applied and the probabilities utilized were acquired from three expert physicians. The cost minimization analysis was performed in three settings. At first, drugs that had a holding time (pH>4) of 11h or more were included. Secondly, drugs that had a holding time of 12h or more were included, and thirdly, a holding time of 13h or more was required. In the first analysis, the least expensive PPI treatment was lansoprazole (average cost of 138.89 per patient). In the second analysis, least expensive treatment was rabeprazole (193.81 per patient), and in the third, rabeprazole again (193.81 per patient). Esomeprazole and omeprazole were not among two of the least expensive alternatives in any of the settings. Which proton pump therapy turns out to be the least expensive for GERD, depends on the length of the holding time desired. Varying the holding time of the drug had a profound effect on the conclusions about the cost-effectiveness of the alternative treatments. PMID- 14757489 TI - The use of formulation technology to assess regional gastrointestinal drug absorption in humans. AB - The aim of this study was to assess the feasibility of using oral modified release formulations for the purposes of site-specific targeting and regional drug absorption assessment in man. An immediate release pellet formulation containing ranitidine as the model drug of choice for the study was fabricated by extrusion-spheronisation, and then film coated with either the enteric polymer polyvinyl acetate phthalate or the bacteria-degradable polymer amylose, in combination with ethylcellulose, to effect drug release within the small intestine and colon, respectively. Optimised formulations were evaluated in vivo in ten healthy volunteers, who each received, on four separate occasions, the immediate release, small intestinal release and colonic release formulations (each equivalent to 150mg ranitidine), and an intravenous injection of ranitidine (equivalent to 50mg ranitidine). Blood samples were collected and assessed for ranitidine concentration, and radiolabelled placebo pellets were co-administered with the coated ranitidine pellets to monitor their gastrointestinal transit using a gamma camera. Ranitidine was rapidly released and absorbed from the immediate release formulation, whereas the enteric formulation (10% coat weight gain) delayed drug release until some or all of the pellets had emptied into the small intestine. The amylose-ethylcellulose coated formulation (coat ratio 1:3, coat weight gain 25%) retarded ranitidine release until the pellets had reached the colon. The mean absolute bioavailability of ranitidine from the immediate release, small intestinal release and colonic release formulations were 50.6, 46.1 and 5.5%, respectively. These data are in general agreement to those obtained from a previous regional intubation study. The present study therefore demonstrates the practical potential of utilising a non-invasive, formulation based approach to assess drug absorption from different regions of the human gastrointestinal tract. PMID- 14757490 TI - Adsorption of the decapeptide Cetrorelix depends both on the composition of dissolution medium and the type of solid surface. AB - High performance liquid chromatography (HPLC) analysis of increasing amounts of the decapeptide Cetrorelix, a potent antagonist of the luteinising hormone releasing hormone, in distilled water resulted in a poor and variable response when solutions of low concentration (0.2-4microg/ml) were analysed. Rinsing experiments revealed loss of analyte due to adsorption to the vial surfaces as the main reason for this. The adsorption of Cetrorelix was found to follow a Langmuir isotherm reaching a plateau at 0.4microg/cm(2) and to be influenced by both the dissolution medium and the type of vial used. The adsorption tendency of Cetrorelix was reduced by: (a) a more lipophilic solvent (ethanol), (b) a more acidic pH (acetic acid) inducing repulsive charges (c) a micellar solution of various tensides. With all of these media the HPLC response was higher (up to five times) and less variable. Adsorption of Cetrorelix to solid surfaces decreased in the rank order: glass > polypropylene = polyethylene > poly (tetrafluoroethylene), with considerable differences between the glass vials of various suppliers. PMID- 14757491 TI - "Apparent" Young's elastic modulus and radial recovery for some tableted pharmaceutical excipients. AB - The effects of compact size and of powder particle size on the determination of "apparent" compressive Young's modulus of elasticity, E, were evaluated, for three pharmaceutical excipients (Microcrystalline cellulose, MCC, Calcium hydrogen phosphate dihydrate, CHPD, and pregelatinized starch, PGS) differing in deformational behaviour during compression. One- and two-compact (composite) specimens were employed and the equations of Spriggs [J. Am. Ceram. Soc. 44 (1961) 628] and Phani & Niyogi [J. Mater. Sci. 22 (1987) 257] were employed for prediction of E, at zero and 0.15 porosity (E(0) and E(0.15)). It was found that E(0) and E(0.15) are affected by the particle size, in the case of PGS, but by the compact size and assembly in the case of MCC and CHPD. Reduction in E(0) and E(0.15) values for two-compact assembly corresponded to large and medium compact size only of MCC and CHPD, while specimen assembly was not affecting significantly the experimental data if small compacts of MCC and CHPD or any size compacts of PGS were used. The exponent f of the Phani and Niyogi equation has been considered as parameter of pore structure and particle morphology. It was significantly affected by the compact size of all the excipients, by the two compact assembly of MCC and CHPD and by the particle size of CHPD and PGS. Absence of significant particle size effect on the parameter f, for the case of MCC is attributed to the elongated particle shape, while the highly significant one for the case of PGS is explained by the extensive elasto-plastic deformation during compression, which is greatly dependent on the grain size. Values of the % radial elastic recovery (%RR), at porosity 0 and 0.15, were determined and correlation of the experimental data was attempted to Young's modulus (E). A simple linear equation is proposed for prediction of Young's modulus, E, from %RR, at least in the porosity range between 0.1 and 0.3 (representative of commercial pharmaceutical tablets), but use of %RR instead to E is applicable only under many limitations. PMID- 14757492 TI - Long-term stabilisation potential of poly(vinylpyrrolidone) for amorphous lactose in spray-dried composites. AB - The aim of this study was to investigate the potential of poly(vinylpyrrolidone) (PVP) to inhibit the crystallisation of amorphous lactose during storage of the composites up to 6 months. Short-term stability was assessed by microcalorimetry over 10 days and long-term stability by storage in desiccators with different relative humidities for 3 and 6 months. The solid-state structure of the particles after storage was analysed by differential scanning calorimetry. It was found that the presence of PVP increased the critical relative humidity (RH) for crystallisation relative to the pure lactose and both the proportion and the molecular weight of the PVP affected the stabilisation of the amorphous phase. The difference in critical RH between the materials increased over time. The T(g) of the materials was generally reduced due to the absorption of water and it is suggested that the inhibiting effect therefore is related mainly to a specific interaction between lactose and PVP, rather than to a counteracting effect of the polymer on the moisture induced depression of T(g). PMID- 14757493 TI - Pharmacokinetic models for the saturable absorption of cefuroxime axetil and saturable elimination of cefuroxime. AB - Since oligopeptidic drugs such as beta-lactam antibiotics share the same carriers in humans and animals, the absorption and elimination kinetics of cefuroxime (C) were investigated in rats. Plasma C concentrations were measured by liquid chromatography. Pharmacokinetics and bioavailability of C in the rat were examined after intravenous (i.v.) administration at three doses (1.78, 8.9 and 17.8mg) of cefuroxime sodium and oral administration at two doses (2.02 and 8.9mg) of cefuroxime axetil (CA). Preliminary fits using data from intravenous administration of C showed that the drug disposition kinetics were clearly nonlinear, with an increase in plasma clearance as the intravenous dose increased. After oral administration of CA, normalized C(max) was higher for smaller dose than for the largest dose. The population pharmacokinetic parameters were obtained by means of nonlinear mixed effect modelling approach according to a nonlinear elimination and nonlinear absorption two-compartment model. The nonlinear elimination could be attributed to a saturable renal tubular reabsorption of the antibiotic and nonlinear intestinal absorption of CA mediated by carrier system. The oral bioavailability of C, calculated by numeric integration of an amount of CA drug absorbed was 22 and 17% for 2.02 and 8.9mg of prodrug administered orally. PMID- 14757494 TI - New 4-aminobicyclo[2.2.2]octane derivatives and their activities against Plasmodium falciparum and Trypanosoma b. rhodesiense. AB - A series of new 2-substituted 4-dialkylaminobicyclo[2.2.2]octane derivatives was prepared and the compounds were investigated for their activity against causative organisms of tropical diseases. The tests were performed as microplate assays using the K1 strain of Plasmodium falciparum (resistant to chloroquine and pyrimethamine) and Trypanosoma brucei rhodesiense (STIB 900). The results were compared to the activities of former tested compounds of the bicyclo[2.2.2]octane series and to known drugs. Most of the 4-amino-6,7-diphenylbicyclo[2.2.2]octan-2 one thiosemicarbazones were compounds with attractive antimalarial potency (IC(50)=0.84-0.99microM, chloroquine: IC(50)=0.12microM). One of the bicyclo[2.2.2]octan-2-yl 4-tert-butylbenzenesulfonates showed the highest antitrypanosomal activity (IC(50)=0.68microM) of the so far prepared 4-amino-6,7 diarylbicyclo[2.2.2]octane derivatives, but is distinctly less active than suramin (IC(50)=0.0075microM). PMID- 14757495 TI - The potential of nasal application for delivery to the central brain-a microdialysis study of fluorescein in rats. AB - Previous animal studies have shown that various types of nasally administered drugs and model substances can access the central nervous system (CNS) via direct transport across the olfactory epithelium, and thereby circumventing the blood brain barrier (BBB). These compounds, however, have mainly been identified in the cerebrospinal fluid and the olfactory bulbs which are usually not pharmacologically relevant targets. The aim of the present study was to evaluate the potential of targeting the central brain by olfactory absorption by use of sodium fluorescein as a hydrophilic model substance with limited permeability across the blood-brain barrier. Microdialysis probes were implanted in blood and in right and left side of the brain (striatum) in rats. The pharmacokinetics of sodium fluorescein was studied from 0 to 180min following intravenous and unilateral nasal administration without occlusion of the oesophagus. Pharmacokinetic modelling showed a significantly higher absorption rate and lower T(max) in the ipsilateral striatum (0.097min(-1) and 41min) compared with the contralateral side (0.056min(-1) and 54min). The rate of elimination in brain was significantly lower after nasal administration (0.004min(-1)) compared with intravenous administration (0.012min(-1)). However, the brain to plasma area under the curve ratios of model substance were low (2-3%) and not significantly different between right and left side of the brain, regardless of the route of administration. The results obtained by microdialysis were supported by findings in whole brain homogenates where concentrations of fluorescein were approximately 40% higher in the right striatum compared with the left side initially after nasal administration to the right nostril of rats. Despite some indications of olfactory transport to the central rat brain it was concluded that the drug targeting potential of sodium fluorescein and most likely other hydrophilic compounds is limited. PMID- 14757496 TI - Membrane effects of the antitumor drugs doxorubicin and thaliblastine: comparison to multidrug resistance modulators verapamil and trans-flupentixol. AB - The interactions of the antitumor drugs doxorubicin and thaliblastine with model membranes composed of neutral (phosphatidylcholine) and negatively charged (phosphatidylserine) phospholipids were studied by differential scanning calorimetry and nuclear magnetic resonance. The membrane activities of doxorubicin and thaliblastine were compared to those of the powerful multidrug resistance (MDR) modulators trans-flupentixol and verapamil. The results point out to the potential role of the drug-membrane interactions for the effects of doxorubicin and thaliblastine in resistant tumor cells. They direct also to the artificial membranes as a suitable tool for screening of compounds with potential ability to modulate MDR. PMID- 14757497 TI - Influence of hydroxypropyl-beta-cyclodextrin complexation on piroxicam release from buccoadhesive tablets. AB - Interaction of piroxicam (PX) and hydroxypropyl-beta-cyclodextrin (HPbetaCD) was investigated in solution and in the solid state. Solubility studies demonstrated the formation of the PX-HPbetaCD inclusion complex with 1:1 stoichiometry. Equimolecular PX-HPbetaCD solid systems were prepared and characterized by differential scanning calorimetry, Fourier transform infrared spectroscopy, and X ray diffractometry. Modification of the release of a sparingly water-soluble drug, PX, from hydrophilic matrices using cyclodextrin complexation was evaluated. The buccoadhesive controlled release tablets for the delivery of PX were prepared by direct compression of hydroxypropylmethyl cellulose (HPMC) and Carbopol 940 (C940), which showed superior bioadhesion properties compared to HPMC. The tablets were evaluated for their dissolution, swelling and mucoadhesive properties. The in vitro release results demonstrated that matrix tablets containing the PX-HPbetaCD solid complex displayed faster PX release compared to those containing a physical mixture or "free" drug. Differences in release rates of PX from the tablets could be attributed to the presence of the polymers and to cyclodextrin complexation. The effect of the polymers on PX release can affect the drug solubility (complexation) and polymer water uptake (swelling). Higher polymer water uptake may result in higher drug solubility and diffusivity in a hydrated polymeric environment. Drug complexation affected also its diffusivity through the semipermeable membrane. PMID- 14757498 TI - Integrated pharmacokinetic-pharmacodynamic modeling and allometric scaling for optimizing the dosage regimen of the monoclonal ior EGF/r3 antibody. AB - The multiple-dose strategy with the monoclonal ior EGF/r3 antibody, in xenograft bearing nude mice, was supported upon the basis of its integrated pharmacokinetic pharmacodynamic relationship, according to both the temporal (K(e0)=0.0015+/ 0.000035h(-1)) and the time-independent sensitivity (C(50%)(ss), 9.23+/ 0.17microg/ml; C(max,eff)(ss), 12.5microg/ml) components of its tumor growth delay action. This relationship was consistent with a sigmoidal E(max) pharmacodynamic model postulating a hypothetical effect compartment that permits us to estimate an effective steady-state concentration range (7.5-12microg/ml). Using this information we calculated both the cumulative and non-cumulative dosage regimens to compare their response patterns with respect to the control group. It follows that the differences in the estimated tumor growth inhibition ratio were statistically significant between the control group and either of the treated ones (P<0.05). The median survival time in treated mice under non cumulative regimen (72+/-10 days), predicted an increase in this parameter as compared to the control one (55+/-6 days). Finally, using the allometric paradigm, the empiric power equation for dose scaling across mammalian species allowed the calculation of the dosage schedule for further clinical trial. The estimated maintenance dose in human (70kg) was 200mg/m(2) to be given weekly, and the corresponding loading dose was 600mg/m(2). PMID- 14757499 TI - Compaction, compression and drug release characteristics of xanthan gum pellets of different compositions. AB - Compaction and compression of xanthan gum (XG) pellets were evaluated and drug release from tablets made of pellets was characterised. Three formulations were prepared by extrusion-spheronisation and included, among other excipients, diclofenac sodium (Dic Na), at 10% (w/w); xanthan gum, at 16% (w/w); and one of three different fillers (lactose monohydrated (LAC), tribasic calcium phosphate (TCP) and beta-cyclodextrin (beta-CD)), at 16% (w/w). Five hundred milligrams of pellets (fraction 1000-1400microm) were compacted in a single punch press at maximum punch pressure of 125MPa using flat-faced punches (diameter of 1.00cm). Physical properties of pellets and tablets were analysed. Dissolution was performed according to the USP paddle method. Pellets showed close compressibility degrees (49.27% LAC; 51.32% TCP; and 50.48% beta-CD) but densified differently (3.57% LAC; 14.84% TCP; 3.26% beta-CD). Permanent deformation and densification were the relevant mechanisms of compression. Fragmentation was regarded as non-existent. The release behaviour of tablets made of pellets comprising LAC or beta-CD was anomalous having diffusional exponent (n) values of 0.706 and 0.625, respectively. Drug diffusion and erosion were competing mechanisms of drug release from those tablets. PMID- 14757500 TI - Reversal of P-glycoprotein mediated vincristine resistance of L1210/VCR cells by analogues of pentoxifylline. A QSAR study. AB - In our previous papers we described the ability of methylxanthine pentoxifylline (PTX) to depress the P-glycoprotein (P-gp) mediated multidrug resistance (MDR) of the mouse leukemic cell line L1210/VCR. Other methylxanthines like caffeine and theophylline were found to be ineffective in this respect. In the present paper we have analysed the capability of 25 methylxanthines to depress MDR of L1210/VCR cells. These methylxanthines structurally differ in substituents located in positions N1, N3, N7 and C8. The results indicate that for an effective reversal of P-gp mediated MDR of our cells the existence of a longer polar substituent in the position N1 plays a crucial role. The elongation of the substituent in the positions N3 and N7 (from methyl to propyl) increases and in the position C8 (from H to propyl) decreases the efficacy of xanthines to reverse the vincristine resistance of L1210/VCR cells. The multiple linear regression for effectiveness of methylxanthines in reversal of P-gp mediated MDR of L1210/VCR cells (expressed as respective IC(50r) values) has been computed, with molar weight: M(w), molar volume: V(M), molar refractivity: R(M), crystal density: d and partition coefficient n-octanol/water: logP as descriptors. A high intercorrelation of M(W), V(M) and R(M) was found for the tested group of methylxanthines indicating that only one of these parameters is necessary for testing a potential correlation. The best fit in the multiple linear regression was obtained for R(M) applied together with d and logP and resulted in a QSAR model given by the following equation: IC(50r)=-[(32.3+/-7.2)x10(-3)xR(M)]+[(10.1+/-2.3)xd]+[(0.74+/ 0.10)xlogP]-[10.5+/-3.2]. Model revealed that: (i) the molar refractivity influences the effectiveness of xanthine positively; (ii) the crystal density and partition coefficient influence the MDR reversal effectiveness of xanthine negatively. PMID- 14757501 TI - Characterization and taste-masking evaluation of acetaminophen granules: comparison between different preparation methods in a high-shear mixer. AB - The aim of this study was to prepare and to investigate acetaminophen taste masked granules obtained in a high-shear mixer using three different wet granulation methods (method A: water granulation, method B: granulation with a polyvinylpyrrolidone (PVP) binding solution and method C: steam granulation). The studied formulation was: acetaminophen 15%, alpha-lactose monohydrate 30%, cornstarch 45%, polyvinylpyrrolidone K30 5% and orange flavour 5% (w/w). In vitro dissolution studies, performed at pH 6.8, showed that steam granules enabled the lower dissolution rate in comparison to the water and binding solution granules; these results were then confirmed by their lower surface reactivity (D(R)) during the dissolution process. Moreover, the results of the gustatory sensation test performed by six volunteers confirmed the taste-masking effects of the granules, especially steam granules (P<0.001). Morphological, fractal and porosity analysis were then performed to explain the dissolution profiles and the results of the gustatory sensation test. Scanning electron microscopy (SEM) analysis revealed the smoother and the more regular surface of steam granules with respect to the samples obtained using methods A and B; these results were also confirmed by their lower fractal dimension (D(s)) and porosity values. Finally, differential scanning calorimetry (DSC) results showed a shift of the melting point of the drug, which was due to the simple mixing of the components and not to the granulation processes. In conclusion, the steam granulation technique resulted a suitable method to comply the purpose of this work, without modifying the availability of the drug. PMID- 14757502 TI - The rationale of scored tablets as dosage form. AB - The aim of the study was to get insight into the rationale of scored tablets. This was pursued by studying patient's reasons for subdividing ("breaking") scored and unscored tablets. Patients who picked up their prescriptions in 5 community pharmacies in The Netherlands were questioned. Two-hundred and seventy five prescriptions were studied. Of all dispensed tablets, 31% were subdivided, mostly because a dose that needed subdivision was prescribed. However, 30% were subdivided upon the initiative of the patient himself: 13% for ease of swallowing and 17% because the patient chose to take a lower dose. Even unscored tablets were subdivided: because the dose prescribed was half the tablet dose (6%), for ease of swallowing (1%) and the wish of the patient to take a lower dose (3%). It was also inquired about the patient's perception of the ease of subdivision of scored tablets. Problematic subdivision of scored tablets was reported in 55% of the cases, 42% of which was attributed to a disfunctioning score line. We also studied the possibility to prescribe and dispense other medicinal products as alternatives for tablets that needed to be subdivided. For 46% a dosage form with a lower dose was on the market, for 54% it was not. We conclude that scored tablets still fulfil an important role. Even when lower dosed tablets would become available, there remains a substantial wish of patients to subdivide tablets for ease of swallowing and adapting the dose. Improving the functioning of score lines may be a more practical approach than banning this dosage form. PMID- 14757503 TI - Carrier-mediated transport of clonidine in human keratinocytes. AB - The alpha-2 agonist clonidine is frequently used in transdermal therapeutic systems for antihypertensive therapy. This study was performed to characterize transport of clonidine into human keratinocytes. The uptake of [3H]clonidine was measured into monolayers of the human cell line HaCaT and normal human epidermal keratinocytes in primary culture. The uptake of clonidine was linear for up to 1min, independent of Na(+), but pH-dependent. Uptake was carrier-mediated with an affinity constant (K(t)) of 0.30mM and a maximal velocity (V(max)) of 15.7nmol/min per mg of protein. Diphenhydramine, guanabenz, procainamide, tryptamine, quinine, and quinidine, but not choline markedly inhibited clonidine uptake. We conclude that clonidine is transported into keratinocytes in a pH dependent manner by a saturable uptake system different from the keratinocyte choline transporter. The substrate specificity of the system corresponds to that of the recently characterized system for tertiary amines. After diffusion of the drugs through the stratum corneum, this transport system might contribute to the passage of clonidine and diphenhydramine across the living epidermis after dermal administration. PMID- 14757504 TI - Alpha-tocopherol influences the lipid membrane affinity of desipramine in a pH dependent manner. AB - Phopholipidosis is a lipid storage disorder caused by cationic amphiphilic drugs (CADs) characterized by the lysosomal accumulation of phospholipids and drug. alpha-Tocopherol (alpha-Toc) has a reversible effect on phospholipidosis in rats and cell culture. We studied the influence of alpha-Toc on the partitioning of the CAD desipramine in a liposome/buffer system using equilibrium dialysis with the following lipid compositions: egg phosphatidylcholine (PhC) or wheat germ phosphatidylinositol (PhI) or a combination of PhC, PhI and cholesterol, containing between 1.5 and 20% (mol per mol total lipids) of alpha-Toc, alpha tocopherol acetate (alpha-TocAc), 2,2,5,7,8-pentamethyl-6-chromanol (PMC) or cholesterol. alpha-Toc (1.5%) enhanced the partition coefficient of neutral desipramine by up to 1.1 log units while it had no influence on the partitioning of the ionized compound. In the PhC liposome system, at pH 7.4 logD increased with increasing alpha-Toc concentrations but was unchanged at pH 4.5. Similar effects were found with PMC while alpha-TocAc or cholesterol, between 1.5 and 20%, had no influence on the partitioning of desipramine. From these results we postulate that in vivo, alpha-Toc could mediate a redistribution of CADs from lysosomal membranes (pH approximately 4.5) to membranes and lipoproteins at physiological pH. PMID- 14757505 TI - Synthesis, and antiprotozoal and antibacterial activities of S-substituted 4,6 dibromo- and 4,6-dichloro-2-mercaptobenzimidazoles. AB - The synthesis and some germicidal activities in vitro of two congener series of S substituted 4,6-dihalogeno-2-mercapto-1H-benzimidazoles are reported. There was no substantial difference between antibacterial activities of corresponding 4,6 dichloro- and 4,6-dibromo-derivatives. The present results confirm lower susceptibility to substituted benzimidazoles of Gram-negative compared to Gram positive bacteria. Minimum inhibitory concentrations (MICs) of a majority of the novel derivatives ranged between 25 and 100microg/ml for Gram-positive bacteria. The most active compounds (MICs for Gram-positive bacteria: 0.78-50microg/ml) were 4,6-dichloro-2-(4-nitrobenzylthio)-1H-benzimidazole and 4,6-dibromo-2-(4 nitrobenzylthio)-1H-benzimidazole that were 4-32 times more potent than nitrofurantoin against all Gram-positive bacteria utilized but Escherichia faecalis, against which they were, respectively, 2 and 4 times less potent than nitrofurantoin. Among Gram-negative bacteria used, Stenotrophomonas maltophilia and Bordetella bronchiseptica were most sensitive (as evidenced by a number of MICs 400microg/ml). All the new compounds were at least several times more active against Giardia intestinalis (IC(50): 0.006-0.053microg/ml), and a half of them were at least several times more active against Trichomonas vaginalis (IC(50): 0.0015-0.182microg/ml) than metronidazole (IC(50): 0.210 and 0.037microg/ml, respectively), the drug of choice in the treatment of G. intestinalis and T. vaginalis infections. PMID- 14757506 TI - A common pathway of nitric oxide release from AZD3582 and glyceryl trinitrate. AB - 4-(Nitrooxy)-butyl-(S)-2-(6-methoxy-2-naphthyl)-propanoate (AZD3582) is a cyclooxygenase (COX)-inhibiting nitric oxide donator (CINOD). It donates nitric oxide (NO) in biological systems through as yet unidentified mechanisms. cGMP, a marker of intracellularly generated NO, was increased up to 27-fold over basal levels by AZD3582 (1-30microM) in LLC-PK1 kidney epithelial cells. A 5h pretreatment with glyceryl tinitrate (GTN, 0.1-1microM) attenuated the cGMP response to a subsequent challenge with AZD3582 or GTN. Similarly, AZD3582 (10 30microM) pretreatment reduced the increase in cGMP on subsequent incubation with AZD3582 or GTN. In contrast, cGMP stimulation by SIN-1, which releases NO independently of enzymatic catalysis, remained unimpaired in cells pretreated with GTN or AZD3582. Our results demonstrate that AZD3582 decreases the sensitivity of the guanylyl cyclase/cGMP system to GTN and vice versa. This suggests that bioactivation pathways for organic nitrates, which involve enzymatic catalysis, may be responsible for NO donation from AZD3582. PMID- 14757507 TI - Effect of heat on the percutaneous absorption and skin retention of three model penetrants. AB - The effect of heat on the transdermal delivery of model penetrants of differing lipophilicity through artificial membranes (non-rate limiting) and human epidermis was investigated in vitro. Saturated suspensions of the model penetrants; methyl paraben (MP), butyl paraben (BP) and caffeine (CF) in deionised water (vehicle) were used to attain maximal thermodynamic activity. Franz cell experiments were performed at temperatures ranging from 23 to 45 degrees C using the infinite dose method. Artificial membrane studies showed the penetrant diffusivity (diffusion coefficient) in the vehicle to be totally dependent on temperature and not changes in donor solubility. Epidermal flux and retention of all penetrants was found to be affected by temperature. The amount of penetrant retained in the epidermis was found to be in the order BP>CF>MP whilst the transdermal fluxes increased in the order MP>BP>CF with increasing receptor temperature. Estimated epidermal diffusivity of MP was found to be significantly affected by temperature (P< or =0.05) compared to BP and CF. Using Arrhenius plots, a lower activation energy was recorded for CF and may suggest a difference in permeation kinetics compared to the other penetrants. PMID- 14757508 TI - Transport properties and association behaviour of the zwitterionic drug 5 aminolevulinic acid in water. A precision conductometric study. AB - The behavior of the hydrochloride salt of 5-aminolevulinic acid (ALA-HCl) with respect to transport properties and dissociation in aqueous solution at 25 degrees C has been studied using precision conductometry within the concentration range 0.24-5.17mM. The conductivity data are interpreted according to elaborated conductance theory. The carboxyl group appears to be, in practice, undissociated. The dissociation constant, K(a), of the NH(3)(+) form of the amino acid molecules is determined to 6.78x10(-5) (molarity scale); pK(a)=4.17. The limiting molar conductivity of the ALA-H(+) ion, lambda(0)=33.5cm(2)Omega(-1)mol(-1); electric mobility u=3.47x10(-4)cm(2)V(-1)s(-1), is close to the electric mobilites of the acetate and benzoic ions. PMID- 14757509 TI - Assessment of chitosan derivatives as buccal and vaginal penetration enhancers. AB - The aim of the present work was to evaluate the mucoadhesive and penetration enhancement properties via the buccal and vaginal mucosae of four different chitosan derivatives: 5-methyl-pyrrolidinone chitosan (MPC), two low molecular weight chitosans (DC1 and DC2) and a partially reacetylated chitosan (RC). Chitosan HCl was used as a reference. Polymer solutions (4% w/w) were prepared in media simulating the buccal (pH 6.4 buffer or water) and the vaginal (pH 5.0 buffer) environments and subjected to rheological characterization. Acyclovir was added to the polymer solutions at 5% (w/w) concentration. The mucoadhesive properties of the polymer solutions were measured using excised porcine cheek or vaginal mucosa and mucin dispersions to simulate the buccal or vaginal environments, respectively. Drug permeation and penetration tests were carried out using porcine cheek and vaginal mucosae as model membranes. Acyclovir aqueous suspensions prepared in pH 6.4 and 5.0 buffers were used as blanks. Drug release measurements were also carried out in the same conditions employed for the permeation and penetration tests. Methyl-pyrrolidinone chitosan shows the best mucoadhesive and penetration enhancement properties in both buccal and vaginal environments. The capability to enhance the permeation/penetration of acyclovir was decreased by partial depolymerization of chitosan and disappeared after partial reacetylation. PMID- 14757510 TI - Pure antiestrogen RU 58668-loaded nanospheres: morphology, cell activity and toxicity studies. AB - Nanospheres (NS) formulated using biodegradable and biocompatible polymers, poly(D,L-lactide-co-glycolide) (PLGA), poly(D,L-lactide) (PLA) and poly(epsilon caprolactone) (PCL), loaded with the pure anti-estrogen RU 58668 (RU), a promising estrogen-dependent anticancer agent, have been prepared. They all possess a small size compatible with an intratumoral extravasation behavior and their pegylation reduce significantly their zeta potential. Characterization by freeze fracture electron microscopy have shown that NS are spheric particles with a size ranging between 30 and 50nm and a tendency to agglomerate which is reduced by polyethylene glycol (PEG) grafting. PEG-grafted NS are all non-toxic as revealed by cell viability assay. A specific cellular model has been used to evaluate not only the release extent of the drug but also its biological activity. All formulations tested showed that they release slowly RU as measured by the delayed ability of RU to inhibit estrogen-induced transcription in human breast cancer cells and that they possess only a small amount of surface adsorbed RU. PMID- 14757511 TI - Delivery and stability of LHRH and Nafarelin in human skin: the effect of constant/pulsed iontophoresis. AB - Poor absorption and stability of peptides are the major obstacles concerning the development of therapeutically relevant iontophoretic devices for the transdermal delivery of peptides. The present study examined the impact of constant and pulsed (direct/alternating) current profiles on the transport and stability of two decapeptides LHRH and Nafarelin. The stability of these peptides was studied in a physiological buffer solution, with electrical current, and when the peptide solution was exposed to the stratum corneum or to the epidermal/dermal side of human skin. Pulsed direct current profile was shown to be the most efficient in transporting both LHRH and Nafarelin across the human epidermis. Furthermore, the percentage of intact LHRH in the receiver phase was slightly higher when a pulsed current profile was used. Both the peptides were stable in a physiological buffer and under the influence of current, but LHRH was degraded especially in contact with the dermal side of the skin. Altogether five hydrolytic degradation products of LHRH were observed, and they were identified by LC-ESI/MS and LC-ESI/MS/MS. No degradation products of Nafarelin were observed. It is concluded that the pulsed direct current profile may provide at least a partial solution for the transdermal delivery of peptides in terms of improved transport efficacy and peptide stability. PMID- 14757512 TI - Differentiated in vivo skin penetration of salicylic compounds in hairless rats measured by cutaneous microdialysis. AB - The purpose was to investigate the in vivo skin penetration of four 14C-salicylic compounds using microdialysis and to relate dermal concentrations to structural features. Furthermore, to compare two in vivo retrodialysis recovery methods for estimation of true unbound extracellular concentrations. Microdialysis probes were inserted in the dermis of hairless rats. Equimolal 14C-salicylic formulations were applied topically and dialysate sampled consecutively for 4h. True extracellular concentrations were estimated by retrodialysis by drug method (the 14C-salicylic compounds themselves) and by retrodialysis by calibrator method (3H-salicylic acid as internal standard). Probe depth was measured by ultrasound scanning. High dermal concentrations were found after application of 14C-salicylamide (low protein-binding) and the lipophilic ester 14C-butyl salicylate, which was completely hydrolysed to 14C-salicylic acid during skin diffusion. Protein binding and dissociation may explain the lower dermal concentrations of 14C-salicylic acid and 14C-diethylamine salicylate, respectively. Probe depth did not significantly influence dialysate concentrations. The two in vivo recovery correction methods did not reduce the variation in concentration-time curves. In conclusion, differentiated penetration kinetics was found ranking: 14C-salicylamide >/= 14C-butyl salicylate > 14C salicylic acid > 14C-diethylamine salicylate. Dermal concentrations were related to structural features of the model compounds. The two correction methods performed alike; however, the calibrator method has the advantage of serving as a quality control during experiments. PMID- 14757513 TI - Developmental changes of keratin expression in chick embryo olfactory epithelium in relation to cellular differentiation and neurogenesis in vivo and in vitro. AB - Olfactory embryogenesis was studied using an anti-chick keratin antibody on chick embryo sections as well as in vitro. Olfactory placodes form at embryonic day 3 (ED3) in the anterior facial ectoderm and invaginate to form the nasal pits. At ED5, the epidermal ectoderm and respiratory epithelium show the same dense cytokeratin immunoreaction. In contrast, absence of keratin expression in the basal part of olfactory epithelial primordium, in the deeper nasal pit area, coincides with one of the critical first steps of olfactory neurogenesis. However, beginning with periphero-central olfactory synaptogenesis at ED8, a new basal cell population starts to express keratin in the olfactory epithelium. Keratin positive cells appear to correspond, by their epithelial localisation and morphology, to sustentacular and basal cells. This interpretation was confirmed in vitro with ED14 chick primary olfactory cultures where TrKA immunoreactivity was used as a marker of horizontal basal cells (HBCs). After ED15, late keratin expression was detected in forming Bowman's glands. The density of keratin expressing basal cells was measured between ED10 and ED20, and appeared highest in the median part of the olfactory epithelium, the area of most active olfactory neurogenesis and neuronal maturation. Thus, keratin expression corresponds to a specialisation of horizontal basal cells as active neuronal stem cells. PMID- 14757514 TI - Alterations in synaptic transmission and plasticity in area CA1 of adult hippocampus following developmental hypothyroidism. AB - Transient reductions in thyroid hormone during critical periods of brain development can have devastating and irreversible effects on neurological function. The hippocampus is a brain region sensitive to thyroid hormones and is a necessary substrate for some forms of learning and memory. Subregions within the hippocampus display distinct ontogenetic profiles and have shown differential vulnerability to some indices of thyrotoxic insult. Synaptic function can be readily assessed in the hippocampus, yet little information exists on the consequences of early thyroid hormone insufficiency on the neurophysiological integrity of this structure. Previous work has examined the long-term consequences of perinatal hypothyroidism on neurophysiology of the dentate gyrus of the hippocampal formation. The current study reveals that alterations in synaptic function also exist in area CA1, and some differences in the pattern of effects are evident between the two hippocampal subfields. Developing rats were transiently exposed to the thyrotoxicant, propylthiouracil (PTU; 0 or 15 ppm), through the drinking water of pregnant dams beginning on gestational day 18. This regimen markedly reduced circulating levels of thyroid hormones and stunted pup growth. PTU exposure was terminated on postnatal day (PN) 21 and electrophysiological assessments were conducted by recording field potentials in area CA1 of hippocampal slices derived from adult male offspring. Synaptic transmission, short-term, and long-term synaptic plasticity were assessed. Consistent with observations in the dentate gyrus, somatic population spike amplitudes were reduced in assessments of baseline synaptic transmission of slices from PTU-exposed animals. No differences were identified in excitatory postsynaptic potentials (EPSP). Short-term plasticity of the EPSP as indexed by paired pulse facilitation was markedly impaired by PTU exposure. Long-term potentiation (LTP) of the population spike was enhanced, consistent with findings in dentate gyrus, but no change in EPSP LTP was detected. Perturbations in synaptic function in the hippocampus of adult rats transiently exposed to a period of hormone insufficiency during the perinatal period are likely to contribute to cognitive deficits associated with developmental hypothyroidism. PMID- 14757515 TI - Mouse outer hair cells lacking the alpha9 ACh receptor are motile. AB - Efferent nerve fibers form chemical synapses at the bases of outer hair cells (OHC), with acetylcholine (ACh) being their principal neurotransmitter. The activation of ACh receptors on OHCs is known to influence cochlear function. These efferent effects exhibit an unusual pharmacology and are generally known to be inhibitory. Recent evidence suggests that an ACh receptor subunit, known as alpha9, plays a dominant role in mediating the olivocochlear neurotransmission to OHCs. In this investigation, we attempt to determine the possible role(s) of the alpha9 subunit in regulating OHC function by examining OHC electromotility and compound action potentials (CAP) in mice carrying a null mutation for the alpha9 gene. Results indicate that cochlear sensitivity, based on CAP thresholds, is similar for homozygous mutant and wild-type mice. Electromotility is also present in OHCs, independent of whether the alpha9 subunit is present or absent. PMID- 14757516 TI - An early Fgf signal required for gene expression in the zebrafish hindbrain primordium. AB - We have explored the role of fibroblast growth factor (Fgf) signaling in regulating gene expression in the early zebrafish hindbrain primordium. We demonstrate that a dominant negative Fgf receptor (FgfR) construct disrupts gene expression along the entire rostrocaudal axis of the hindbrain primordium and, using an FgfR antagonist, we find that this Fgf signal is required at early gastrula stages. This effect cannot be mimicked by morpholino antisense oligos to Fgf3, Fgf8 or Fgf24--three Fgf family members known to be secreted from signaling centers at the midbrain-hindbrain boundary (MHB), in rhombomere 4 and in caudal mesoderm at gastrula stages. We propose that an Fgf signal is required in the early gastrula to initiate hindbrain gene expression and that this is distinct from the later roles of Fgfs in patterning the hindbrain during late gastrula/early segmentation stages. We also find that blocking either retinoic acid (RA) or Fgf signaling disrupts hindbrain gene expression at gastrula stages, suggesting that both pathways are essential at this stage. However, both pathways must be blocked simultaneously to disrupt hindbrain gene expression at segmentation stages, indicating that these signaling pathways become redundant at later stages. Furthermore, exogenous application of RA or Fgf alone is sufficient to induce hindbrain genes in gastrula stage tissues, suggesting that the two signal requirement can be overcome under some conditions. Our results demonstrate an early role for Fgf signaling and reveal a dynamic relationship between the RA and Fgf signaling pathways during hindbrain development. PMID- 14757517 TI - Chlorpyrifos exposure during neurulation: cholinergic synaptic dysfunction and cellular alterations in brain regions at adolescence and adulthood. AB - The developmental neurotoxicity of chlorpyrifos (CPF) involves multiple mechanisms, thus rendering the immature brain susceptible to adverse effects over a wide window of vulnerability. Earlier work indicated that CPF exposure at the neural tube stage elicits apoptosis and disrupts mitotic patterns in the brain primordium but that rapid recovery ensues before birth. In the current study, we assessed whether defects in cholinergic synaptic activity emerge later in development. CPF was given to pregnant rats on gestational days 9-12, using regimens devoid of overt maternal or fetal toxicity. We then examined subsequent development of acetylcholine systems and compared the effects to those on general biomarkers of cell development. Choline acetyltransferase (ChAT), a constitutive marker for cholinergic nerve terminals, was increased in the hippocampus and striatum in adolescence and adulthood. In contrast, hemicholinium-3 (HC-3) binding to the presynaptic choline transporter, an index of nerve impulse activity, was markedly subnormal. Furthermore, m2-muscarinic cholinergic receptor binding was significantly reduced, instead of showing the expected compensatory upregulation for reduced neural input. CPF also elicited delayed-onset alterations in biomarkers of cell packing density, cell number, cell size and neuritic projections, involving brain regions both with and without reductions in indices of cholinergic activity. In combination with earlier results, the current findings indicate that the developing brain, and especially the hippocampus, is adversely affected by CPF regardless of whether exposure occurs early or late in brain development, and that defects emerge in adolescence or adulthood even in situations where normative values are initially restored in the immediate post exposure period. PMID- 14757518 TI - Sex differences in rapid auditory processing deficits in microgyric rats. AB - Early neocortical injury has been associated with rate-specific auditory processing deficits using rodent models. In the few cases where females were studied, they appeared less vulnerable than males to the behavioral consequences of early neocortical injury. In the current study, male rats with neocortical microgyria were found to exhibit significant impairments in detecting tone sequences at short but not long inter-stimulus intervals (ISI) as compared to sham-operated male littermates. Microgyric females, however, performed similarly to sham-operated female littermates on this task at all durations. Current findings support an association between focal cortical malformations and impaired rapid auditory processing in males, and less vulnerability in females to the behavioral consequences of these malformations on a task eliminating confounds of motivation, experience, and estrus. PMID- 14757519 TI - Vascular endothelial growth factor promotes neurite maturation in primary CNS neuronal cultures. AB - Recent studies have demonstrated that vascular endothelial growth factor (VEGF) and its receptor VEGFR2 (flk-1) are expressed by neurons during development and following hypoxic-ischemic events. Moreover, fetal CNS tissue explants exposed to exogenous VEGF exhibit increased neuronal Map-2 expression, suggesting that VEGF could have an effect on neuronal maturation. To determine whether this effect is of a direct nature, we examined the expression of Map-2 in the presence of VEGF in primary CNS neuronal cultures. After 3 days in culture, a statistically significant dose-dependent increase in the length of Map-2(+) processes was observed, with the peak occurring at 10 ng/ml of VEGF. Immunohistochemical analysis of the cultures demonstrated the presence of VEGFR2 after VEGF treatment, as well as the expression of the VEGF receptor VEGFR1 (flt-1). Treatment of the cultures with antisense oligonucleotides against VEGFR2, but not against VEGFR1, abolished the effect of VEGF on the length of Map-2(+) processes. RT-PCR analyses of Map-2 and VEGFR1 indicated that mRNAs of these two genes are upregulated in the presence of VEGF. The addition of wortmannin, an inhibitor of PI3K/Akt signal-transduction pathway, to the media did not affect the VEGF dependent increase in Map-2(+) length. In contrast PD98059, which inhibits the MAPK pathway, partially abolished this effect of VEGF. These experiments suggest that VEGF has a direct effect on neuronal growth and maturation under normoxic conditions during CNS development, which is mediated by the VEGFR2 receptor via the MAPK pathway. PMID- 14757520 TI - Immunocytochemical expression of monocarboxylate transporters in the human visual cortex at midgestation. AB - Lactate and the other monocarboxylates are a major energy source for the developing brain. We investigated the immunocytochemical expression of two monocarboxylate transporters, MCT1 and MCT2, in the human visual cortex between 13 and 26 post-ovulatory weeks. We used immunoperoxidase and immunofluorescence techniques to determine whether these transporters co-localized with markers for blood vessels (CD34), neurons (microtubule-associated protein 2 [MAP2], SMI 311), radial glia (vimentin), or astrocytes (glial fibrillary acidic protein [GFAP], S100beta protein). MCT1 immunoreactivity was visible in blood vessel walls as early as the 13th week of gestation mainly in the cortical plate and subplate. At this stage, less than 10% of vessels in the ventricular layer expressed MCT1, whereas all blood vessels walls showed this immunoreactivity at the 26th gestational week. Starting at the 19th week of gestation, sparse MCT1 positive cell bodies were detected, some of them co-localized with MAP2 immunoreactivity. MCT2 immunoreactivity was noted in astrocytic cell bodies from week 19 and spread subsequently to the astrocyte end-feet in contact with blood vessels. MCTs immunoreactivities were most marked in the subplate and deep cortical plate, where the most differentiated neurons were located. Our findings suggest that monocarboxylate trafficking between vessels (MCT1), astrocytes (MCT2) and some postmitotic neurons (MCT1) could develop gradually toward 20 gestational weeks (g.w.). These data suggest that lactate or other monocarboxylates could represent a significant energy source for the human visual cortex at this early stage. PMID- 14757521 TI - Cellular expression of P2Y and beta-AR receptor mRNAs and proteins in freshly isolated astrocytes and tissue sections from the CA1 region of P8-12 rat hippocampus. AB - Although almost all GFAP(+) cells in primary astrocyte cultures show functional beta-adrenergic (beta-AR) and metabotropic purinergic (P2Y) receptors, the fewer studies on astrocytes in situ have shown that a much smaller proportion express these same receptor-mediated activities. Here we show, by multiplex single cell RT-PCR, that 44% of freshly isolated, GFAP(+) astrocytes (FIAs) from the CA1 of P8-12 rat hippocampus always co-express beta-adrenergic receptor mRNA subtypes with metabotropic ATP receptor mRNA subtypes (P2Y1, P2Y2 or P2Y4). We also found that beta2 mRNA was the dominant beta-AR subtype expressed. P2Y1 mRNA always co expresses with either one or two subtypes of P2U-like receptor (P2Y2 or P2Y4) mRNAs. Immunocytochemical studies showed a similar percentage of all FIAs expressed beta-AR and P2Y1 protein (54% and 52%, respectively), as for the mRNAs (46% and 65%, respectively). The staining of hippocampal sections for beta-AR or P2Y1 receptor plus GFAP shows that there are quite numerous, scattered star shaped GFAP(+) astrocytes in the CA1 region of P9-10 rat hippocampus that stained positive for either of these receptors. These data show that astrocytes in situ express, and to a large extent likely co-express, beta-AR and P2Y receptors. PMID- 14757522 TI - Reelin-expressing neurons in the anterior commissure and corpus callosum of the rat. AB - Reelin is an extracellular matrix protein, which plays a crucial role for the formation of laminated and nonlaminated structures in the central nervous system. To elucidate its roles in the postnatal brain, in the present study, we raised a polyclonal antibody specific for rat Reelin, and investigated Reelin-expressing neurons in the rat brain during the postnatal periods in detail. We found that some Reelin-expressing cells existed in the anterior commissure and corpus callosum. These Reelin-expressing cells were also immunostained with the antibody specific for neurons, but not immunostained with the antibodies specific for astrocytes nor oligodendrocytes, suggesting that these Reelin-expressing cells in the white matter are neurons. They are also immunostained with anti-GAD67 antibody, indicating that Reelin-expressing cells in the commissure systems are GABAergic neurons. Reelin-expressing neurons in the anterior commissure had many conspicuous varicosities on their dendritic arbors and mimic to the interfascicular neurons. These results suggest that Reelin may participate in the regulatory mechanism of neuronal activities through the commissure structure during the postnatal periods. PMID- 14757523 TI - Synaptic lesions and synaptophysin distribution change in spinal motoneurons at early stages following sciatic nerve transection in neonatal rats. AB - The purpose of this investigation was to evaluate the rate of synaptic stripping, changes in the synaptophysin distribution, and synapses ultrastructure of spinal motoneurons at early stages of sciatic nerve axotomy in newborn rats. Seven groups were used in the experiment, which were sacrificed after 1, 3, 6, 12, 24, 48 and 72 h. L4-L6 spinal segments from the animals of the above groups were prepared and processed for indirect immunoperoxidase. Accordingly, tissue samples were prepared from similar groups for routine electron microscopy. Synaptophysin labeled motoneurons were classified into intact, partial, cytoplasmic and negative patterns. Synaptophysin immunoreactivity in both the axotomized and non axotomized sides showed no negative pattern at the first three time points, while this pattern significantly increased in the 12, 24, 48 and 72 h groups at the axotomized side, moreover, there was a progressive reduction in the percentage of the intact pattern at the axotomized side. The percentages of the cytoplasmic and partial patterns also increased during the time course. The rate of synaptic stripping was five time higher in the axotomized side than that of non-axotomized side. The results of the ultrastructural study showed synaptic membranes irregularity, synaptic vesicles displacement, synaptic membrane detachment and ensheathment of degenerated synapses. The conclusion of the study was that early synaptophysin immunoreactivity changes were seen in both the axotomized and non axotomized sides, but the rate of change in the axotomized side was more rapid than that of the non-axotomized side. PMID- 14757524 TI - Development of circadian rhythmicity and photoperiodic response in subdivisions of the rat suprachiasmatic nucleus. AB - To ascertain whether the circadian rhythmicity of the ventrolateral (vl) suprachiasmatic nucleus (SCN) develops concurrently with that of the dorsomedial (dm) SCN and when the rhythmicity starts to respond to day length, i.e., to the photoperiod, rats with their offspring were maintained under either a long photoperiod with 16 h of light and 8 h of darkness per day (LD 16:8) or under a short, LD 8:16 photoperiod. The rhythms of spontaneous c-Fos immunoreactivity in the dm-SCN and of the light-induced c-Fos immunoreactivity in the vl-SCN were studied in the pups. In 3- and 10-day-old rats, the dm-SCN rhythm in spontaneous c-Fos immunoreactivty was already well expressed but a response to a photoperiod similar to that in adult rats has not yet been developed. The vl-SCN gate for insensitivity of c-Fos production to light at certain times was detected in 10 day but not yet in 3-day-old rats: in the latter, light exposure at any daytime induced high c-Fos immunoreactivity. In the 10-day-old pups, similarly as with adult rats, the gate was shorter under LD 8:16 than under LD 16:8, but the difference in the gate duration between the short and the long photoperiod did not yet attain that of adult animals. The data indicate that the circadian rhythmicity may develop sooner in the dm-SCN, than in the vl-SCN, whereas the photoperiodic response may develop sooner in the vl-SCN. PMID- 14757525 TI - The effect of prenatal hypoxia and malnutrition on memory consolidation in the chick. AB - The contribution of hypoxia and malnutrition to cognitive impairments was investigated in chicks incubated in conditions of reduced gas exchange. Previous research has shown that reducing gas exchange during incubation by wrapping half the eggshell with an impermeable membrane results in impaired cognitive ability in young chicks. The results were interpreted within a three stage sequential model of memory using discriminated bead avoidance learning. Reducing gas exchange for 4 days from day 10 or 14, of the 21-day incubation, inhibits memory formation and consolidation into permanent storage. The nature of the cognitive deficit depended on the timing of the insult. Environmental hypoxia (14% oxygen), induced from days 10 to 14 and from days 14 to 18, replicated the memory deficits found previously when eggs were partially wrapped with a membrane. Oxygen is necessary to break down food and to provide energy to build tissue proteins, and therefore hypoxia (partial wrapping or environmental incubation) may indirectly cause malnutrition. Malnutrition, induced by removing 5%, 7.5% or 10% albumin from the egg prior to incubation, had no significant effect on memory consolidation. Raised corticosterone levels occurred in chicks malnourished by 5% and 7.5%, but brain sparing was only evident in chicks with 7.5% albumin removal. Hatch rates were very low in 10% malnourished chicks. Using the chick as a model of prenatal stress, we have been able to isolate the effects of hypoxia from contributing maternal factors. PMID- 14757527 TI - Postnatal development of the alpha1 containing GABAA receptor subunit in rat hippocampus. AB - Here we have studied the developmental expression of alpha1 subunit of the GABAA receptor in comparison with the expression of alpha2 subunit and several GABAergic markers (parvalbumin (PV), calretinin (CR), somatostatin (SOM), neuropeptide Y (NPY) and vasoactive intestinal polypeptide (VIP)). The alpha1 expression (mRNA and protein) was low at birth and increased progressively until the adulthood. This expression pattern was similar to that observed for PV, opposite to that of CR (high at birth and decreased continuously until the adulthood) and differed from that observed for the alpha2 and neuropeptides (SOM, NPY and VIP) (in all cases, a clear peak in expression was observed at P10). We further investigated the expression of alpha1, PV and CR by immunohistochemistry. As expected, the alpha1 and the PV expression were low at birth and increased progressively until the adulthood. Both alpha1 and PV were co-expressed by the same interneuronal population, however, the maturation of the alpha1 subunit preceded to that of PV. Finally, we observed a gradient of maturation between the different fields of the hippocampus proper (CA2-3 preceded to CA1 and DG). This gradient could be related to the high expression of CR positive cells and fibers during the first 10 postnatal days, located principally in the stratum lacunosum moleculare of the CA2-3 layers. PMID- 14757526 TI - Receptor-like protein tyrosine phosphatase zeta/RPTP beta is expressed on tangentially aligned neurons in early mouse neocortex. AB - Protein tyrosine phosphatase zeta (PTPzeta)/RPTPbeta is a chondroitin sulfate proteoglycan predominantly expressed in the brain. In this study, we examined immunohistochemical localisation of PTPzeta in the mouse telencephalon from embryonic day 9.5 (E9.5) to E15.5. During E10.5-E12.5, immunoreactivities for PTPzeta are specifically observed on the tangentially aligned neurons at the preplate (PP) of the neocortex, as well as on the neurons at the mantle layer (ML) of the ganglionic eminences (GEs). Likewise, neurons immunoreactive for CR50, a marker for Cajal-Retzius neurons, are aligned from the ML of the ganglionic eminences to the PP of the neocortex and co-express PTPzeta. During E13.5-E15.5, PTPzeta-positive neurons are present at the subplate (SP) as well as at the marginal zone (MZ) of the neocortex. These results indicate that PTPzeta is a useful marker for early-generated neocortical neurons in mice: Cajal-Retzius neurons as well as the subplate neurons. PMID- 14757528 TI - Patterns of developmental mRNA expression of neurturin and GFRalpha2 in the rat striatum and substantia nigra do not suggest a role in the regulation of natural cell death in dopamine neurons. AB - We examined the mRNA expression of neurturin (NTN) and its receptor GFRalpha2 in rat substantia nigra (SN) and striatum by northern analysis at ages ranging from postnatal day (PND) 2 to adult. NTN mRNA expression is developmentally regulated in striatum with a peak at PND10, but its expression in striatum is low, and less than that of SN. In SN, there is no developmental regulation. GFRalpha2 was expressed most highly during the first two postnatal weeks. Like NTN, GFRalpha2 mRNA was also more abundant in SN, at both PND2 and 14. Our results show that NTN expression is relatively low in the striatum, the target of dopamine (DA) neurons, and there is no apparent pattern of developmental regulation in SN. Thus these studies are not strongly supportive of a role for NTN in regulating natural cell death (NCD) in DA neurons, either as a target-derived or as a local paracrine factor. PMID- 14757530 TI - Oncogenes as therapeutic targets. PMID- 14757529 TI - Regulation of glutamate receptor RNA editing and ADAR mRNA expression in developing human normal and Down's syndrome brains. AB - In human brain, developmental up-regulation in RNA editing at the Q/R site was evident in GluR5 and GluR6, but GluR2 editing in the white matter was down regulated. Each ADAR mRNA expression was up-regulated in the gray matter, whereas differently regulated in the white matter. ADAR2 mRNA was not overexpressed in the brains of Down's syndrome subjects, nor was there any evidence of changes in the RNA editing efficiency of their GluRs. PMID- 14757531 TI - Conditional animal models: a strategy to define when oncogenes will be effective targets to treat cancer. AB - The ability to model cancer in the mouse has provided a robust methodology to dissect the molecular etiology of cancer. These models serve as potentially powerful platforms to preclinically evaluate novel therapeutics. In particular, the recent development of strategies to conditionally induce the or knockout the function of genes in a tissue specific manner has enabled investigators to engineer mice to demonstrate that the targeted inactivation of specific oncogenes can be effective in inducing sustained regression of tumors. Thus, these animal models will be useful to define the specific genes that will be therapeutically useful to target for the treatment of particular human cancers. PMID- 14757532 TI - New approaches in identifying drugs to inactivate oncogene products. AB - With an information explosion on the molecular mechanism of oncogenesis, the completion of the human genome sequence project, and the advances in genomic and proteomic methods, many therapeutic targets for various cancers have been identified. It is timely that a number of new drug development techniques have been developed in this last decade. Candidate drug targets can now be efficiently validated with RNA interference and transgenic animals studies. Combinatorial chemistry provides large numbers of chemical compounds for drug lead discovery and optimization. High throughput assays and high content cell-based assays, in conjunction with sophisticated robotics, are now available for screening large numbers of compounds. Based on X-ray crystallographic structure data, drug leads can be discovered through in silico screening of virtual libraries. By applying these various drug discovery techniques, it is anticipated that more potent and specific anti-cancer agents will be discovered within the next decade. PMID- 14757533 TI - Targeted treatment of hypereosinophilic syndromes and chronic eosinophilic leukemias with imatinib mesylate. AB - Idiopathic hypereosinophilic syndrome (HES) and chronic eosinophilia leukemia (CEL) represent the most recent additions to the list of molecularly defined chronic myeloproliferative disorders. Beginning with the observation that imatinib mesylate (Gleevec) could elicit rapid and complete hematologic remissions in a proportion of patients with HES, a reverse bedside-to-bench translational research effort led to the discovery of FIP1L1-PDGFRA, a novel fusion gene on chromosome 4q12 whose product is an imatinib-sensitive protein tyrosine kinase. FIP1L1-PDGFRA is the first description of a gain-of-function fusion gene derived from an interstitial chromosomal deletion rather than a reciprocal translocation. Empiric use of imatinib in HES and CEL provides a dramatic example of how the development of targeted therapeutics can provide tremendous insight into the molecular etiology of what appear to be a diverse and otherwise indecipherable collection of diseases. In this review, we discuss the role of imatinib in HES/CEL and other malignancies characterized by constitutively activated tyrosine kinases, and examine molecular features of the FIP1L1-PDGFRA fusion. PMID- 14757534 TI - 'Targeting' the epidermal growth factor receptor tyrosine kinase with gefitinib (Iressa) in non-small cell lung cancer (NSCLC). AB - Gefitinib (ZD1839, Iressa), a selective drug inhibitor of the epidermal growth factor receptor (EGFR) tyrosine kinase (TK), was recently approved for the treatment of patients with advanced non-small cell lung cancer (NSCLC). This article reviews the identification of EGFR as a therapeutic target and the steps in the development of gefitinib as "targeted" monotherapy for patients with NSCLC. Whether EGFR is required for the maintenance of lung tumor survival is also discussed. Finally, strategies to identify predictors of response to gefitinib are explored. PMID- 14757535 TI - Targeted therapies in myeloid leukemia. AB - Targeted therapies for hematological malignancies have come of age since the advent of all trans retinoic acid (ATRA) for treating APL and STI571/Imatinib Mesylate/Gleevec for CML. There are good molecular targets for other malignancies and several new drugs are in clinical trials. In this review, we will concentrate on individual abnormalities that exist in the myelodysplastic syndromes (MDS) and myeloid leukemias that are targets for small molecule therapies (summarised in Fig. 1). We will cover fusion proteins that are produced as a result of translocations, including BCR-ABL, the FLT3 tyrosine kinase receptor and RAS. Progression of diseases such as MDS to secondary AML occur as a result of changes in the balance between cell proliferation and apoptosis and we will review targets in both these areas, including reversal of epigenetic silencing of genes such as p15(INK4B). PMID- 14757536 TI - Targets for molecular therapy of skin cancer. AB - Cancers of the skin encompass the first and second most common neoplasms in the United States, epidermal basal cell carcinoma (BCC) and squamous cell carcinoma (SCC), respectively, as well as the melanocytic malignancy, malignant melanoma (MM). Recently identified alterations in the function of specific genes in these cancers provide new potential therapeutic targets. These alterations affect conserved regulators of cellular proliferation and viability, including the Sonic Hedgehog, Ras/Raf, ARF/p53, p16(INK4A)/CDK4/Rb and NF-kappaB pathways. New modalities designed to target these specific proteins may represent promising approaches to therapy of human skin cancers. PMID- 14757537 TI - Can mouse models for brain tumors inform treatment in pediatric patients? AB - Brain tumors represent the most common solid tumor of childhood. Although the histology of many pediatric brain tumors is similar to that of their adult counterparts, significant differences exist with regard to tumor location and response to therapy. The biological and genetic basis for this difference is poorly understood, as tumor tissue is generally unavailable for such studies. While targeted therapies directed against specific molecules active in cancer represents a new arsenal of agents for treating these tumors, such agents are generally not being developed for pediatric cancer in particular. Therefore, new agents for treatment of pediatric glioma must be obtained from compounds being tested against tumors of comparable histology in adult patients. Compounding this problem, although brain tumors are among the most lethal tumors of childhood, their absolute number is relatively small. As a consequence, trials with new agents must be prioritized based on the likelihood that a particular agent or combination of agents will have efficacy in pediatric cancer. Mouse models for brain tumors may help to identify targeted agents, and combinations of agents, effective against these tumors. Such data can be used to prioritize therapies for clinical trials in children with these tumors. PMID- 14757538 TI - Human exposure monitoring and evaluation in the Arctic: the importance of understanding exposures to the development of public health policy. AB - Arctic indigenous peoples face significant challenges resulting from the contamination of Arctic air, water, and soil by persistent organic pollutants, heavy metals, and radionuclides. International cooperative efforts among governments and research institutions are under way to collect the information needed by environmental health scientists and public health officials to address environmental contamination in the Arctic. However, the climatic, political, and cultural conditions of the land and its native populations combine to present a unique set of scientific and logistic challenges to addressing this important public health issue. Public health officials have the responsibility to respect the cultural traditions of indigenous communities, while simultaneously designing strategies that will reduce their exposure to environmental contaminants and rates of disease and dysfunction. Researchers can better understand the link between environmental exposures and disease through monitoring programs for both the subsistence diets and health status of the indigenous populations. We suggest that the incorporation of community-based participatory research methods into programs designed to assess biomarkers of contaminant exposure in children and adults may be a valuable addition to ongoing and newly developed research programs. This approach could serve as a model for international environmental health initiatives, because it involves the participation of the local communities and seeks to builds trust between all stakeholders. PMID- 14757541 TI - The World Health Report 2003: conveying new insights while refurbishing old ideas. PMID- 14757542 TI - The HIV/AIDS pandemic: at last good news. PMID- 14757543 TI - The impact of income: assessing the relationship between income and health in Sweden. AB - AIMS: This paper explores the relationship between income and health among adults in Sweden. An analysis was made as to what extent the association differs when one studies individual earnings and equivalent disposable income, as well as gender differentials. Further, a study was undertaken to investigate how, and by what magnitude, the income-health relationship changes when one controls for other structural factors, such as education and class. Finally the functional form of the relationship was scrutinized, because of its obvious policy impact. METHODS: Data came from the 1996-97 Swedish Living Condition Surveys, which include individuals aged 25-64 (n=7,201). Logistic regression was used, including various polynomial terms of the income variable. RESULTS: The results show that both earnings and disposable household income are strongly related to health, a finding that holds for both women and men. The strength of the association becomes somewhat weaker when one controls for other structural factors, but in the final model the association is in fact about the same as the bivariate association, owing to the impact of age. Moreover, a curvilinear association was revealed by the authors' analyses. CONCLUSIONS: A clear association was found between income and health, also when other structural variables are controlled for. This indicates that income, as such, is of great importance for the risk of illness. The shape of the association between income and health is consistent with earlier debates concerning the relation between income distribution and population health indicators, and, as such, indicates that income-equalizing policies may have an impact on health. PMID- 14757544 TI - Systematic medical data collection of intentional injuries during armed conflicts: a pilot study conducted in West Bank, Palestine. AB - AIMS: A study was undertaken on implementing medical data collection as a tool to assess the relative number and character of intentional injuries before and during an armed conflict. METHODS: Data on hospital contacts due to intentional injury were collected retrospectively at two hospitals in West Bank, Palestine, and classified by ICD10 and the new International Classification of External Causes of Injuries (ICECI). A three-month period prior to the current Intifada, June-August 2000 and two three-month periods of the Intifada, September-December 2000 and June-August 2001 were chosen. RESULTS: The number of contacts increased from 23 in the period prior to the Intifada to 740 during the first and 199 during the second period of the Intifada. During the period before the Intifada the victims were men, and no one was younger than 10 years. During the Intifada 9% were women, 3% below 10 years, and 9% aged 10-14 years. Prior to the Intifada all injuries were caused by blunt force. During the Intifada 65% of the injuries were caused by firearms or explosives, 19% by beating and 6% by gaseous substances. Among children most lesions were localized to the head, including eye and brain damage, and were mostly caused by firearms. CONCLUSIONS: Injury registration by ICD10 combined with ICECI codes facilitates analyses of correlations between characteristics of armed conflicts and injuries. Medical data collection is an important instrument in documentation of the effects of weapons and in surveillance of violations of humanitarian law, particularly as to the worrying magnitude of young children being seriously victimized. PMID- 14757545 TI - The new age of the molecular family: an anthropological view on the medicalisation of kinship. AB - Now that genetic inheritance is featuring more and more as an explanation of disease and human behaviour in general, a question that needs to be asked is how such explanations affect people's perceptions of family and kinship and to what extent genetic explanations conflict with broader social developments. Ideas about the genetic inheritance of disease place the family and kin group in the spotlight, requiring all its members to be scrutinised. Research on inheritable diseases entails a medicalization of kinship that reflects and promotes a view of family relationships at odds with the ongoing changes in the structure of families. At a time when family structures are more fluid and less determined by "blood" relationships than ever before we have an increasing emphasis on genetic inheritance as the transmitter of both human behaviour and kinship. Embedded in concepts of genetic inheritance is thus the notion that family and kin are the medium through which inheritance flows. Two cases from an anthropological study will illustrate how genetic mapping leads to the medicalization of kinship. PMID- 14757546 TI - Developing the formula for state subsidies for health care in Finland. AB - AIM: The aim was to generate a research-based proposal for a new subsidy formula for municipal healthcare services in Finland. METHODS: Small-area data on potential need variables, supply of and access to services, and age-, sex- and case-mix-standardised service utilisation per capita were used. Utilisation was regressed in order to identify need variables and the cost weights for the selected need variables were subsequently derived using various multilevel models and structural equation methods. RESULTS: The variables selected for the subsidy formula were as follows: age- and sex-standardised mortality (age under 65 years) and income for outpatient primary health services; age- and sex-standardised mortality (all ages) and index of overcrowded housing for elderly care and long term inpatient care; index of disability pensions for those aged 15-55 years and migration for specialised non-psychiatric care; and index of living alone and income for psychiatric care. CONCLUSION: Decisions on the amount of state subsidies can be divided into three stages, of which the first two are mainly political and the third is based on the results of this study. PMID- 14757547 TI - Illicit drug abuse in second-generation immigrants: a register study in a national cohort of Swedish residents. AB - AIMS: This study investigates ethnic and socioeconomic risk factors for hospital admissions related to illicit drug abuse in second-generation immigrants in Sweden. METHOD: Cox analyses of proportional hazards were used to estimate the relative risk of sociodemographic covariates in analyses of register data on hospital admissions because of illicit drug abuse during 1990-99. The study population was a national cohort of 1.25 million residents (aged 10-30 years). RESULTS: Second-generation immigrants had two- to three fold higher age and sex adjusted relative risks (RRs) for hospital admissions because of illicit drug use compared with the Swedish majority population with a limited variation between different ethnic groups. The RRs decreased greatly after the model was adjusted for socioeconomic indicators of the childhood household. Inter-country adoptees had the highest risk for hospital admission related to illicit drug abuse of all study groups after adjustment for sociodemographic variables (RR 2.8). CONCLUSIONS: Second-generation immigrants are at particular risk for illicit drug abuse in Sweden. Adverse socioeconomic living conditions are very important in explaining this high risk. PMID- 14757548 TI - Correlates of probable alcohol abuse among women working in nursing homes. AB - AIMS: The purpose of this nationwide study was to assess the prevalence of probable alcohol abuse (PAA) among women working in geriatric care, and to study its demographics, medical and work-related correlates. METHODS: The employees of geriatric nursing homes and geriatric hospital wards in Iceland with 10 patients or more were invited to participate in this cross-sectional study. The response rate was 80% (n=1515), with 96% being women (n=1432). Men were consequently omitted from the study. Questions were included on demographics, psychosocial factors, workplace environment, health behavior, and medical history. PAA was defined as (a) having been given such a diagnosis by a physician, (b) having missed work because of drinking, or (c) if alcohol use was considered a problem by the employee herself, her family, friends, or the employer. RESULTS: A total of 4.8% of the employees fulfilled the criteria for PAA. These women were younger (41 vs. 45 years of age), more often single (25% vs. 15%) or divorced (13% vs. 9%), and less satisfied with work than the other women. Odds ratios for asthma, fibromyalgia, chronic fatigue, pain syndromes, mental disorders, and work-related accidents were elevated. Despite this, no differences were found concerning amount of sick leave. Their psychosocial work environment was worse but the physical work environment was the same. CONCLUSION: Women with probable alcohol abuse working in nursing homes have significant medical problems and psychosocial morbidity that is not reflected in more sick leave. PMID- 14757549 TI - Emergency room utilization in Copenhagen: a comparison of immigrant groups and Danish-born residents. AB - BACKGROUND: The aim of the study was to investigate whether utilization of the emergency room differed between immigrant groups and Danish-born residents in Copenhagen, Denmark. METHODS: The authors compared the number of emergency room contacts during 1997 among different ethnic groups in the study population. Data were provided by the Statistical Office of the Municipality of Copenhagen. The study population consisted of 183,478 citizens residing in the catchment area of Bispebjerg Hospital in Copenhagen on 1 January 1998. "Contacts" included 22,026 visits made to the emergency room at Bispebjerg Hospital during 1997. Both the study population and "contacts" were characterized by gender, age, income, and country of birth. The immigrants comprised nine ethnic groups according to country of birth. Data were analysed by Poisson regression comparing rate ratios. RESULTS: Persons born in Somalia, Turkey, and ex-Yugoslavia had higher utilization rates of emergency room than Danish-born residents. All other non Western-born residents had utilization rates similar to Danish-born residents. Persons born in other Western and European countries showed a tendency towards less utilization. CONCLUSION: Higher utilization rates among some immigrant groups may be explained by disparities in health or lack of knowledge about the Danish healthcare system as well as barriers to seeking primary care including language, fear of discrimination, and low satisfaction with primary care. The challenge remains to identify these causal relations, and to find out why utilization patterns vary between immigrant groups. PMID- 14757550 TI - Physical inactivity in a country in transition: a population-based survey in Tirana City, Albania. AB - AIMS: This paper describes the work and leisure-time physical activity levels of adults living in Tirana City (Albania). METHODS: A cross-sectional survey was undertaken in Tirana City in mid-2001. It included 1120 adults aged 25 years and over (response rate=72.7%). RESULTS: Overall, 19.3% of male and 28.4% of female respondents reported a low level of physical activity at work. During leisure time, 49.5% of men and 57.6% of women (age-standardized prevalence [adjusted to the adult population of Tirana]: 43.4% and 51.9% for men and women respectively) said that they participated in only sedentary activities such as reading and watching television. The gender difference in the likelihood of low physical activity at work and during leisure time was significant even after adjusting for age (age adjusted odds ratios (OR)=1.86 99% confidence interval (CI) 1.12; 3.09 and OR=1.60 99% CI 1.17; 2.22 respectively). The likelihood of leisure-time sedentarity increased with age in both genders but it did not vary significantly with education level or income. Being sedentary during leisure time was also not significantly associated with other unhealthy lifestyle behaviours, including smoking, consumption of vegetables less than six days per week, and daily alcohol intake. CONCLUSIONS: In a country such as Albania that has undergone a rapid transition from a largely unmechanized society, characterized inevitably by high levels of exercise, the generalized high prevalence of low physical activity level during leisure time gives cause for concern regarding future health trends. PMID- 14757552 TI - An "American dilemma" in Scandinavian childbirth: unmet needs in healthcare? PMID- 14757551 TI - The importance of a minimum age law for the possibility of purchase of tobacco by adolescents: a study based on Swedish experiences. AB - AIMS: The purpose of the study is partly to compare the possibility of adolescents purchasing tobacco before and after the introduction of the minimum age law of 18 years, respectively, and partly to examine the factors that characterize the situations in which adolescents may or may not purchase tobacco, respectively. METHODS: Under controlled conditions, adolescents of varying ages carried out test purchases of tobacco. A total of 1,500 attempted purchases were made with the help of 24 adolescents in three regions of Sweden. In 1996, the year before a minimum-age law of 18 for tobacco purchases went into effect, 750 attempted purchases were made, followed by 750 attempted purchases in 1999. RESULTS: In 1996, 91% of the attempted purchases of tobacco were successful. In 1999, the corresponding proportion was 82%. The attempted purchases in 1996 and 1999 show a very strong relationship between whether an age check was made and the result of the attempted purchase. CONCLUSIONS: The study shows that the introduction of a minimum-age law apparently has had a certain effect on the possibility of adolescent purchasing tobacco but that there are obvious shortcomings in the enforcement of the minimum age. The most apparent shortcoming concerns the lack of age controls. This article discusses measures for strengthening age controls in the sale of tobacco to adolescents. PMID- 14757554 TI - An approach to the surgical pathology of tumours and tumour-like conditions of the liver. AB - Hepatic masses are increasingly being found, often as a result of more frequent and sophisticated imaging. Lesions can be detected during screening for primary and metastatic tumours, or as an incidental finding. Although some have distinctive radiological appearances allowing for a confident diagnosis, histological assessment of biopsy and resection specimens remains the cornerstone for the correct identification of many lesions. This broadsheet describes the distinctive features of common and uncommon hepatic tumours and outlines diagnostic problems and pitfalls, particularly for the benign and malignant tumours of hepatocytes and bile ducts. Tumour-like lesions that mimic malignancy are also discussed. Finally, an approach to the assessment of biopsies from probable metastatic tumours is presented, as this is one of the most common indications for liver mass biopsy for the practising pathologist. PMID- 14757555 TI - The role of molecular studies in lymphoma diagnosis: a review. AB - Lymphoma classification is based on a multiparametric approach to diagnosis, in which clinical features, morphology, immunophenotype, karyotype and molecular characteristics are important to varying degrees. While in most cases, a diagnosis can be confidently established on the basis of morphology and immunophenotype alone, a small proportion of diagnostically difficult cases will rely on molecular studies to enable a definitive diagnosis. This review discusses the various molecular techniques available including Southern blotting (SB), polymerase chain reaction (PCR), fluorescence in situ hybridisation (FISH)- including multicolour-FISH/spectral karyotyping and comparative genomic hybridisation--and also gene expression profiling using cDNA microarray technology. Emphasis is given to the analysis of antigen receptor gene rearrangements and chromosomal translocations as they relate to lymphoma diagnosis and also in the setting of minimal residual disease (MRD) detection and monitoring. Laboratories performing these tests need to have expertise in these areas of testing, and there is a need for greater standardisation of molecular tests. It is important to know the sensitivity and specificity of each test as well as its limitations and the pitfalls in the interpretation of results. Above all, results of molecular testing should never be considered in isolation, and must always be interpreted in the context of clinical and other laboratory data. PMID- 14757556 TI - Validation of assembled nucleic acid-based tests in diagnostic microbiology laboratories. AB - Medical microbiology and virology laboratories use nucleic acid tests (NAT) to detect genomic material of infectious organisms in clinical samples. Laboratories choose to perform assembled (or in-house) NAT if commercial assays are not available or if assembled NAT are more economical or accurate. One reason commercial assays are more expensive is because extensive validation is necessary before the kit is marketed, as manufacturers must accept liability for the performance of their assays, assuming their instructions are followed. On the other hand, it is a particular laboratory's responsibility to validate an assembled NAT prior to using it for testing and reporting results on human samples. There are few published guidelines for the validation of assembled NAT. One procedure that laboratories can use to establish a validation process for an assay is detailed in this document. Before validating a method, laboratories must optimise it and then document the protocol. All instruments must be calibrated and maintained throughout the testing process. The validation process involves a series of steps including: (i) testing of dilution series of positive samples to determine the limits of detection of the assay and their linearity over concentrations to be measured in quantitative NAT; (ii) establishing the day-to day variation of the assay's performance; (iii) evaluating the sensitivity and specificity of the assay as far as practicable, along with the extent of cross reactivity with other genomic material; and (iv) assuring the quality of assembled assays using quality control procedures that monitor the performance of reagent batches before introducing new lots of reagent for testing. PMID- 14757557 TI - Genomic alterations in renal tumours: what have we learned in the era of comparative genomic hybridisation? AB - One of the major challenges in cancer research is to generate molecular profiles of tumours and establish correlations between genetic changes and clinical parameters by screening technologies. The identification of tumour-specific gene targets has potential diagnostic and therapeutic implications. Metaphase comparative genomic hybridisation has been used to detect relative DNA-sequence copy number gains (including high-level amplifications of chromosomal regions) and copy number losses in human neoplasms. In the past, metaphase comparative genomic hybridisation has been shown to be a powerful genome-wide screening method and this has considerably advanced our understanding of renal cancer biology. Novel molecular technologies, including array-based comparative genomic hybridisation, fluorescence in situ hybridisation (FISH), cDNA and tissue microarrays will serve to facilitate further characterisation of candidate genes residing in chromosomal regions defined by metaphase comparative genomic hybridisation. This review concentrates on the application of metaphase comparative genomic hybridisation in the area of renal cancer research and summarises data obtained from comparative genomic hybridisation studies. PMID- 14757558 TI - Granular cell tumour: a pitfall in FNA cytology of breast lesions. AB - Granular cell tumours of the breast are rare lesions that may mimic an invasive carcinoma. The fine needle aspiration cytology (FNAC) findings in five cases are presented with specific attention to the pitfalls in the cytological diagnosis of these lesions. PMID- 14757559 TI - Consensus guidelines on anti-cardiolipin antibody testing and reporting. AB - Consensus guidelines on anti-cardiolipin antibody (aCL) testing have been developed to help minimise laboratory variation in the performance and reporting of aCL assays. These guidelines include minimum, optimum and optional recommendations for the following aspects of aCL testing and reporting: (1) isotype of aCL tested; (2) specimen type; (3) controls and assay precision; (4) calibrators; (5) patient samples; (6) rheumatoid factors and IgM aCL testing; (7) reporting of results; (8) cut-off values; and (9) interpretative comments. ABBREVIATIONS: aCL, anti-cardiolipin antibodies; APS, anti-phospholipid antibody syndrome; ASCIA, Australasian Society of Clinical Immunology and Allergy; ASTH, Australasian Society of Thrombosis and Haemostasis; beta2-GPI=beta2-glycoprotein I; ELISA, enzyme-linked immunosorbent assay; NCCLS, National Committee for Clinical Laboratory Standards; HSANZ, Haematology Society of Australia and New Zealand; QAP, Quality Assurance Program; RCPA, Royal College of Pathologists of Australasia; %CV, inter-assay inter-run coefficient of variation. PMID- 14757560 TI - B cell chronic lymphocytic leukaemia cells have reduced capacity to upregulate expression of MHC class I in response to interferon-gamma. AB - AIMS: An important consideration in the design of a tumour vaccine is the ability of tumour-specific cytotoxic T lymphocytes (CTL) to recognise unmanipulated tumour cells in vivo. To determine whether B-CLL might use an escape strategy, the current studies compared B-CLL and normal B cell MHC class I expression. METHODS: Flow cytometry, TAP allele PCR and MHC class I PCR were used. RESULTS: While baseline expression of MHC class I did not differ, upregulation of MHC class I expression by B-CLL cells in response to IFN-gamma was reduced. No deletions or mutations of TAP 1 or 2 genes were detected. B-CLL cells upregulated TAP protein expression in response to IFN-gamma. Responsiveness of B-CLL MHC class I mRNA to IFN-gamma was not impaired. CONCLUSIONS: The data suggest that MHC class I molecules might be less stable at the cell surface in B-CLL than normal B cells, as a result of the described release of beta(2)m and beta(2)m free class I heavy chains from the membrane. This relative MHC class I expression defect of B-CLL cells may reduce their susceptibility to CTL lysis in response to immunotherapeutic approaches. PMID- 14757561 TI - The value of polymerase chain reaction detection of Mycobacterium tuberculosis in granulomas isolated by laser capture microdissection. AB - AIMS: The aim of this study was to investigate the usefulness of polymerase chain reaction (PCR) detection of Mycobacterium tuberculosis in granulomas isolated by laser capture microdissection (LCM). METHODS: The PCR DNA amplification method was used to detect M. tuberculosis in granulomas microdissected from one section stained by haematoxylin and eosin (H&E) from a formalin-fixed paraffin-embedded specimen. The results were compared to those obtained from PCR performed from 10 whole paraffin sections of 5 micro m each, and with the histology, culture and the patient's clinical findings. RESULTS: Forty-nine formalin-fixed and paraffin embedded samples from 49 patients with a histological suspicion of a mycobacterial infection were investigated. Using culture as the reference method, the sensitivity for the detection of M. tuberculosis was 92% and the specificity was 100% using PCR from microdissected granulomas and were similar to those obtained by using PCR from 10 whole sections. CONCLUSIONS: The PCR method of examination of microdissected granulomas from deparaffinised sections is a sensitive, specific and rapid method for the detection of M. tuberculosis in formalin-fixed and paraffin-embedded samples. The method is as sensitive as that using PCR on 10 whole tissue sections, thus making it suitable for small biopsies. However, although these methods reduce the delay in diagnosis, culture remains the gold standard for identification of mycobacteria in tissue. Culture also allows for the testing of antibiotic sensitivity of any isolated species, in this way determining appropriate treatment. PMID- 14757562 TI - Test and teach. An uncommon cause of cutaneous swelling of the thigh. Malignant glomus tumour. PMID- 14757564 TI - Insulin autoimmune syndrome due to IgG kappa paraprotein. PMID- 14757565 TI - Vulvar precancers and cancers in the Northern Territory of Australia. PMID- 14757566 TI - Adenomyolipoma of the endometrium -- a true neoplasm? PMID- 14757567 TI - Polypoid endometrioma of the rectal serosa and retroperitoneal endometriosis in a 70-year-old woman. PMID- 14757568 TI - Anaplastic lymphoma kinase-positive anaplastic large cell lymphoma presenting with spontaneous splenic rupture. PMID- 14757569 TI - Mobile telephones and brain vascular leakage. PMID- 14757570 TI - Re: quantitative evaluation of AgNORs in bone tumours. PMID- 14757575 TI - Impact of ozone on monoterpene emissions and evidence for an isoprene-like antioxidant action of monoterpenes emitted by Quercus ilex leaves. AB - Quercus ilex (L.) leaves emit monoterpenes, particularly alpha-pinene, beta pinene and sabinene. Apart from the monoterpene pools that are stored in specialized structures and have a clear defensive or attractive role, the function of monoterpenes in Q. ilex leaves is unknown. We tested whether monoterpenes have an antioxidant role, as has previously been found for isoprene in isoprene-emitting leaves. We exposed Q. ilex leaves to either mild and repeated ozone exposure (Experiment I) or to a single acute ozone exposure (Experiment II) at temperatures ranging between 20 and 32 degrees C. Both ozone treatments rapidly stimulated monoterpene synthesis, but had no effect on photosynthesis and caused no visible damage to leaves maintained at 25, 30 or 32 degrees C. Ozone inhibited both photosynthesis and monoterpene synthesis in leaves maintained at 20 degrees C. To characterize the relationship between monoterpenes and ozone-induced damage, we fed detached leaves fosmidomycin, a selective inhibitor of isoprene synthesis. Fosmidomycin caused rapid and complete inhibition of monoterpene emissions in leaves maintained at 30 degrees C, confirming that monoterpenes are synthesized by the same biochemical pathway as isoprene. However, over the experimental period, fosmidomycin did not affect concentrations of compounds that are formed from chloroplastic isoprenoids and that might have conferred antioxidant protection, either directly (carotenoids) or indirectly (chlorophylls, xanthophylls). In leaves whose monoterpene synthesis had been inhibited by fosmidomycin, ozone rapidly and significantly inhibited photosynthesis and increased the production of hydrogen peroxide and malonyldialdehyde. We conclude that monoterpenes produced by Q. ilex leaves share the same biosynthetic pathway and function as isoprene. Furthermore, all volatile isoprenoids may have similar antioxidant properties and may be stimulated by the same stress-inducing conditions. PMID- 14757576 TI - Acclimation of photosynthetic capacity in Scots pine to the annual cycle of temperature. AB - Coniferous trees growing in the boreal and temperate zones have a clear annual cycle of photosynthetic activity. A recent study demonstrated that the seasonal variation in photosynthetic capacity of Scots pine (Pinus sylvestris L.) could be attributed mainly to the light response curve of photosynthesis. The magnitude of the light response curve varied over the season while its shape remained constant, indicating that the two physiological parameters quantifying the curve the quantum yield per unit internal carbon dioxide concentration and the corresponding light-saturated rate-remained proportional to each other. We now show, through modeling studies, that the quantum yield (and hence the light saturated rate) is related to the annual cycle of temperature through a delayed dynamic response. The proposed model was tested by comparing model results with intensive measurements of photosynthesis and driving variables made from April to October in three shoots of Scots pine growing near the northern timberline. Photosynthetic capacity showed considerable acclimation during the growing season. A single model describing photosynthetic capacity as a reversible, first order delay process driven by temperature explained most of the variation in photosynthetic capacity during the year. The proposed model is simpler but no less accurate than previous models of the annual cycle of photosynthetic capacity. PMID- 14757577 TI - Abiotic factors limiting photosynthesis in Abies lasiocarpa and Picea engelmannii seedlings below and above the alpine timberline. AB - Most research on the occurrence and stability of alpine timberlines has focused on correlations between adult tree growth and mean temperatures rather than on specific mechanisms. Timberline migration to higher altitude is dependent on new seedling establishment in the tree-line ecotone; however, reductions in photosynthetic carbon gain in establishing seedlings have previously been interpreted solely in terms of decreased seedling survival. Our objective was to evaluate the impact of abiotic factors (temperature, light and water) on photosynthetic carbon gain in young seedlings of the two dominant conifer tree species occurring naturally above (tree-line ecotone site, TS) and below (forest site, FS) a Rocky Mountain timberline in southeastern Wyoming, USA. Coincidentally, measurements were made during an unusually dry summer. Mean daily photosynthesis in seedlings of both Abies lasiocarpa (Hook.) Nutt. (subalpine fir) and Picea engelmannii Parry ex Engelm. (Engelmann spruce) was less at TS than at FS (19 and 29%, respectively). Minimum nighttime temperatures below 2 degrees C were more frequent at TS than at FS and were associated with reduced maximum photosynthesis the following day. Low midday water potentials were associated with a reduction in carbon gain at both sites early in the season, prior to snowmelt, as well as late in the season when soils began to dry. However, the lower photosynthetic rates at TS than at FS appeared to be unrelated to seedling water status because seedlings at both sites had similar xylem pressure potentials. Solar irradiance was highly variable at both sites as a result of uneven shading by neighboring trees, although this variation was substantially reduced on cloudy days (44% of all days observed). Compared with sunny days, cloudy days resulted in greater integrated daily carbon gain at both sites (41% increase at TS and 69% increase at FS), based on a simulated photosynthesis model. Photosynthetic responses to temperature, sunlight and water suggest that variable solar irradiance and nighttime temperatures were major abiotic factors limiting photosynthetic carbon acquisition in these young seedlings, especially for seedlings growing in the tree-line ecotone. PMID- 14757578 TI - Inhibitory effect of flowering and early fruit growth on leaf photosynthesis in mango. AB - Carbohydrate and nitrogen contents, chlorophyll fluorescence and gas exchange were measured in leaves from both vegetative and reproductive terminal shoots of 12-year-old flowering mango trees. Reproductive shoot leaves were close to swelling floral buds, inflorescences or panicles bearing set fruits. Leaves close to inflorescences had lower rates of mitochondrial respiration (Rd) and net photosynthesis (Anet), and lower stomatal conductance (gs) and quantum efficiency of photosystem II under actinic light than vegetative shoot leaves. Leaf nitrogen concentration, which decreased from the beginning until the end of flowering, was lower in leaves close to inflorescences than in vegetative shoot leaves. However, these differences and changes were counterbalanced by an increase in leaf mass-to area ratio so that leaf nitrogen per unit leaf area (Na) remained nearly constant during the whole flowering period, except in leaves close to panicles bearing set fruits. Net CO2 assimilation rate simulated by a biochemical model of leaf photosynthesis (Urban et al. 2003) was much higher than Anet measured at an ambient CO2 partial pressure (Ca) of either 36 or 70 Pa. The overestimation of Anet was more pronounced in leaves close to inflorescences, to panicles bearing set fruits and to reversing inflorescences (characterized by the appearance of leaves in terminal positions on inflorescences) than in vegetative shoot leaves. It is concluded that low Anet in leaves close to inflorescences was probably due neither to changes in Na nor to a decrease in Rubisco activity induced by low gs, but rather to a decrease in electron flow in photosystem II. This decrease was not directly associated with higher starch or soluble sugar contents. PMID- 14757579 TI - Genomic DNA methylation of juvenile and mature Acacia mangium micropropagated in vitro with reference to leaf morphology as a phase change marker. AB - Genomic DNA methylation was analyzed in Acacia mangium Willd. microshoots micropropagated in vitro from juvenile and mature explants, and in relation to leaf morphology of the microshoots, which is considered a phase change indicator. Based on high performance liquid chromatography (HPLC) analyses, we found more DNA methylation in microshoots exhibiting juvenile leaf morphology (22.4%) than in microshoots of the mature phyllode morphological type (20.7%), irrespective of the age of the source material. Overall, the degree of DNA methylation in A. mangium microshoots was consistent with values reported for other angiosperms. Complementary investigations based on methylation sensitive amplification polymorphism (MSAP) techniques established that, of 1204 fragments revealed by the different primer pairs used, 49 (i.e., 4.08%) were derived from C(5m)CGG methylated sites. Three of these C(5m)CGG sites were exclusive to the juvenile plant material, and three sites were exclusive to the mature source. No fragments were associated specifically with leaf morphology, rather than with plant age. Thus, although the two age classes could not be distinguished based on a quantitative HPLC measure of DNA methylation, qualitative differences existed, as demonstrated by the six age-specific markers identified by MSAP. The reliability of the MSAP data was confirmed on a larger sample of juvenile plant material, which suggested that the total of six methylation markers detected is probably an underestimation of the age-related differences in DNA methylation that may exist between juvenile and mature plant materials. PMID- 14757580 TI - Influences of genetic and environmental factors on the concentration of the allergen Cry j 1 in sugi (Cryptomeria japonica) pollen. AB - Pollen from sugi (Japanese cedar, Cryptomeria japonica D. Don), a forest tree species that is widely grown in Japan, causes serious allergic disease. The major allergens from sugi pollen, Cry j 1 and Cry j 2, have been isolated and characterized. It has been reported that Cry j 1 concentration in pollen varies considerably among trees. If Cry j 1 concentration is genetically controlled, the planting of trees with low Cry j 1 concentrations would reduce pollinosis. We investigated genetic and environmental effects on Cry j 1 concentration in eight clones growing at four sites. Concentrations of Cry j 1 in pollen were measured with a monoclonal antibody-based Enzyme Linked Immunosorbent Assay (ELISA). The Cry j 1 concentrations differed significantly among clones and sites, but the site x clone interaction was not significant, suggesting that the Cry j 1 concentration is controlled primarily by genetic factors. We examined correlations between Cry j 1 concentration and temperature and precipitation from July through February. Temperature was not significantly related to Cry j 1 concentration, whereas cumulative precipitation during the 8 months and mean daily precipitation in September showed significant negative correlations with Cry j 1 concentration. PMID- 14757581 TI - Seasonal and annual changes in soil respiration in relation to soil temperature, water potential and trenching. AB - Soil respiration (rs), soil temperature (Ts) and volumetric soil water content were measured in a balsam fir (Abies balsamea (L.) Mill.) ecosystem from 1998 to 2001. Seasonal variation in root and microbial respiration, and covariation in abiotic factors confounded interpretation of the effects of Ts and soil water potential (Psis) on rs. To minimize the confounding effect of temperature, we analyzed the effect of Psis on rs during the summers of 1998-2000 when changes in Ts were slight. Soil respiration declined 25-50% in response to modest water stress (minimum Psis of -0.6 to -0.2 MPa), and between years, there was substantial variation in the relationship between rs and Psis. In the summer of 2000, 2-m2 plots were subjected to drought for 1 month and other plots were irrigated. The relationship between summertime rs and Psis in the experimental plots was similar to that estimated from the survey data obtained during the same summer. In late spring and early autumn of 2001, 2-m2 trenched and untrenched plots were subjected to drought or exposed to rainfall. It was dry in the early autumn and there was severe soil drying (Psis of -10 MPa in untrenched plots and 2 MPa in trenched plots). In spring, rs in untrenched plots responded more to modest water stress than rs in trenched plots, indicating that root respiration is more sensitive than microbial respiration to water stress at this time of year. The response to abiotic factors differed significantly between spring and autumn in untrenched plots but not in trenched plots, indicating that root activity was greater in early autumn than in late spring, and that roots acclimated to the sustained, severe water stress experienced before and during the autumn. PMID- 14757582 TI - Somatic embryogenesis, micropropagation and plant regeneration of "Early Mature" walnut trees (Juglans regia) that flower in vitro. AB - Some walnut trees (Juglans regia L.) originating from central Asia display an early flowering phenotype. These "Early Mature" (EM) trees may produce flowers within months of germination. Secondary flowering waves are also observed within a growing season. Inflorescences may carry male, female and hermaphrodite flowers. Progeny obtained from selected EM trees were cultured in vitro to initiate clonal propagation of these genotypes. Embryogenic lines were established through the culture of immature zygotic embryos. Microshoot lines were obtained from germinated somatic or zygotic embryos. Plants showing EM phenotypes were recovered through direct conversion of somatic embryos or adventitious rooting of microcuttings. During the in vitro propagation phase, flower buds were observed on microshoots after three to six subcultures. Histological analysis showed that most of these flowers were hermaphrodite. In vitro apical buds were used to clone the walnut orthologous cDNAs of the AGAMOUS and APETALA 3 MADS-box genes. Northern blots revealed a preferential expression of both of these homeotic genes in flowers. The results highlight the usefulness of EM lines to study the genetic cues controlling flowering and sexual maturity in woody perennials. PMID- 14757583 TI - Effects of temperature and nutrient availability on plasma membrane lipid composition in Scots pine roots during growth initiation. AB - We studied the effects of root zone temperature (RZT) and nutrient availability on free sterols and phospholipids in the plasma membrane (PM) and on PM-ATPase activity in roots of 1-year-old Scots pine (Pinus sylvestris L.) seedlings during growth initiation in the spring. Seedlings were grown for 6 weeks in hydroponic cultures with low (0.5 mM N; LN) or high (3 mM N; HN) nutrient availability. The root zone was subjected to slow warming (SW) and fast warming (FW) treatments while maintaining similar air temperatures in both treatments. Decreases in the amount of phospholipids and in the phospholipid/free sterol ratio, an increase in the degree of saturation of phospholipid fatty acids and changes in free sterol composition were observed during root growth initiation. Changes in lipid composition of the PM associated with the cold deacclimation process were detected at RZTs above 9 degrees C. Nutrient availability affected the lipid composition of the PM only when RZT was increased slowly. When RZT increased from 4 to 6 degrees C in the SW treatment, the degree of saturation of phospholipid fatty acids decreased, especially in HN seedlings. The sitosterol/stigmasterol ratio remained higher in HN seedlings than in LN seedlings. After an RZT of 9 degrees C had been reached in the SW treatment, HN caused increases in the saturation of phospholipid fatty acids and root PM-ATPase activity, and a decrease in the phospholipid/free sterol ratio. Possible effects of changes in PM lipid composition on root growth and PM-ATPase activity are discussed. PMID- 14757584 TI - Terrestrial plants require nutrients in similar proportions. AB - Theoretical considerations based on nutrition experiments suggest that nutrient ratios of terrestrial plants are similar to the Redfield ratio found in marine phytoplankton. Laboratory experiments have shown that seedlings of many different plant species have similar nutrient concentration ratios when supplied with nutrients at free access. However, at free access, nutrients are likely to be taken up in amounts in excess of a plant's requirements for growth. In further experiments, therefore, the supply rate of each nutrient was reduced so that excessive uptake did not occur. Again, similar nutrient ratios were found among the plant species tested, although the ratios differed from those found in plants given free access to nutrients. Based on the law of the minimum, we suggest that optimum nutrient ratios be defined as the ratios found in plants when all nutrients are limiting growth simultaneously. The literature on nutrient concentrations was surveyed to investigate nutrient ratios in terrestrial ecosystems. Nutrients taken into consideration were nitrogen, phosphorus, potassium, calcium and magnesium. Based on the assumption that nitrogen is either the limiting nutrient or, when not limiting, is taken up only in small excess amounts, we calculated nutrient ratios from published data. The calculated ratios corresponded closely to the ratios determined in laboratory and field experiments. PMID- 14757585 TI - Seasonal changes in gene expression at the sapwood-heartwood transition zone of black locust (Robinia pseudoacacia) revealed by cDNA microarray analysis. AB - Heartwood is a determining factor of wood quality and understanding the biology of heartwood may allow us to control its formation. Heartwood formation is a form of senescence that is accompanied by a variety of metabolic alterations in ray parenchyma cells at the sapwood-heartwood transition zone. Although senescence has been studied at the molecular level with respect to primary growth, the cell maturation and death events occurring during heartwood formation have been difficult to study because of their location and timing. Analysis of global gene expression patterns during the transition from sapwood to heartwood may offer a powerful means of identifying the mechanisms controlling heartwood formation. Previously, we developed cDNA microarrays carrying 2567 unigenes derived from the bark/cambium region, sapwood and transition zone of a mature black locust tree. Here, we describe the use of these microarrays to characterize seasonal changes in the expression patterns of 1873 genes from the transition zone of mature black locust trees. When samples collected in summer and fall were compared, 569 genes showed differential expression patterns: 293 genes were up-regulated (> twofold) in summer (July 5) and 276 genes were up-regulated in fall (November 27). More than 50% of the secondary and hormone metabolism-related genes on the microarrays were up-regulated in summer. Twenty-nine out of 55 genes involved in signal transduction were differentially regulated, suggesting that the ray parenchyma cells located in the innermost part of the trunk wood react to seasonal changes. We established the expression patterns of 349 novel genes (previously unknown or no-hit), of which 154 were up-regulated in summer and 195 were up-regulated in the fall. PMID- 14757586 TI - Seasonal variation of soil respiration in a Pinus cembra forest at the upper timberline in the Central Austrian Alps. AB - Soil respiration (R) of a 95-year-old Pinus cembra L. forest at the alpine timberline was measured continuously from October 2001 to January 2003 with an automated multiplexing gas exchange system. There was significant spatial variability in soil respiration, and R at a soil temperature of 10 degrees C (R10) decreased by about 20% m(-1) with increasing distance from the trunk. Needle litter and fine root density also decreased. The spatially averaged annual soil CO2 efflux was 35 g C m(-2) year(-1) in 2002. About 70% of the temporal variation in soil respiration could be explained by variations in soil temperature, whereas the influence of soil water potential and thus soil water content was negligible because soil water availability was supra-optimal. PMID- 14757588 TI - Acquiring and inhibiting prepotent responses in schizophrenia: event-related brain potentials and functional magnetic resonance imaging. AB - BACKGROUND: Schizophrenia is associated with deficits in using context to establish prepotent responses in complex paradigms and failures to inhibit prepotent responses once established. OBJECTIVE: To assess prepotent response establishment and inhibition in patients with schizophrenia using event-related brain potential (ERP) and functional magnetic resonance imaging (fMRI) in a simple NoGo task. To combine fMRI and ERP data to focus on fMRI activations associated with the brief (approximately 200 ms) moment of context updating reflected in the NoGo P300 ERP component. DESIGN AND SETTING: We collected ERP and fMRI data while subjects performed a NoGo task requiring a speedy button press to X stimuli (P=.88) but not to K stimuli (P=.12). The ERPs were collected at the Veterans Affairs Palo Alto Health Care System, Palo Alto, Calif; fMRIs were collected at Stanford University, Stanford, Calif. PARTICIPANTS: We recruited patients with DSM-IV schizophrenia (n=11) from the community and the VA hospital and sex- and age-matched healthy control subjects (n=11) from the community. MAIN OUTCOME MEASURES: Behavioral accuracy, P300 amplitudes and latencies, and fMRI activations suggested that patients with schizophrenia did not establish as strong a prepotent tendency to respond to the Go stimulus as healthy subjects. In healthy subjects, NoGo P300 was related to activations in the anterior cingulate cortex, dorsal lateral prefrontal cortex, and right inferior parietal lobule and caudate nucleus, perhaps reflecting conflict experienced when withholding a response, control needed to inhibit a response, and stopping a response in action, respectively. In patients with schizophrenia, NoGo P300 was modestly related to activations in the anterior cingulate cortex, which is consistent with experiencing conflict. CONCLUSIONS: The difference in ERP and fMRI responses to Go and NoGo stimuli suggested that inhibiting a response was easier for patients with schizophrenia than for healthy subjects. Correlations of P300 and fMRI data suggested that patients with schizophrenia and healthy subjects used different neural structures to inhibit responses, with healthy subjects using a more complex system. PMID- 14757589 TI - Presynaptic dopaminergic dysfunction in schizophrenia: a positron emission tomographic [18F]fluorodopa study. AB - CONTEXT: The dopamine overactivity hypothesis of schizophrenia remains one of the most influential theories of the pathophysiology of the illness. Radiotracer brain imaging studies are now directly testing aspects of the overactivity hypothesis. OBJECTIVE: To assess presynaptic dopaminergic function in a large cohort of patients with schizophrenia by means of [18F]fluorodopa uptake and a high-sensitivity 3-dimensional positron emission tomograph. We predicted elevations in striatal [18F]fluorodopa uptake and reductions in prefrontal cortical [18F]fluorodopa uptake in patients with schizophrenia. DESIGN: Case control study. SETTING: Research institute investigation recruiting hospital outpatients. PATIENTS: Sixteen male medicated hospital outpatients with a DSM-IV diagnosis of schizophrenia (mean age, 38 years) and 12 age-matched male volunteers free of psychiatric and neurologic illness. INTERVENTION: [18F]fluorodopa positron emission tomographic scanning. MAIN OUTDOME MEASURE: [18F]fluorodopa uptake constant Ki measured with statistical parametric mapping and region-of-interest analyses. RESULTS: Statistical parametric mapping (P<.05 corrected) and region-of-interest analyses (P<.01) showed increased [18F]fluorodopa uptake, confined primarily to the ventral striatum in patients with schizophrenia. No reductions in prefrontal cortical [18F]fluorodopa uptake Ki were seen in the statistical parametric mapping and region-of-interest analyses, although dorsal anterior cingulate [18F]fluorodopa Ki correlated with performance on the Stroop Color-Word Test in both groups. CONCLUSIONS: As in studies in unmedicated patients, presynaptic striatal dopamine dysfunction is present in medicated schizophrenic patients, adding further in vivo support for dopamine overactivity in the illness. PMID- 14757590 TI - Reducing the duration of untreated first-episode psychosis: effects on clinical presentation. AB - CONTEXT: Most studies on first-episode psychosis show an association between a long duration of untreated psychosis (DUP) and poorer short-term outcome, but the mechanisms of this relationship are poorly understood. OBJECTIVE: To determine whether it is possible to reduce the DUP for first-episode patients in a defined health care area through the introduction of an early detection (ED) program, compared with parallel health care areas without an ED program (No-ED). SETTING AND PATIENTS: We included consecutive patients with a DSM-IV diagnosis of nonorganic, nonaffective psychosis coming to their first treatment in the study health care areas between January 1, 1997, and December 31, 2000. A total of 281 patients (76% of the total) gave informed consent. INTERVENTIONS: The ED and No ED health care areas offered an equivalent assessment and treatment program for first-episode psychosis. The ED area also carried out an intensive ED program. RESULTS: The DUP was significantly shorter for the group of patients coming from the ED area, compared with patients from the No-ED areas (median, 5 weeks [range, 0-1196 weeks] vs 16 weeks [range, 0-966 weeks]). Clinical status measured by the Positive and Negative Syndrome Scale and the Global Assessment of Functioning Scale was significantly better for patients from the ED area at start of treatment and, with the exception of Positive and Negative Syndrome Scale positive subscale, at 3 months. Multiple linear regression analyses gave no indication that confounders were responsible for these differences. CONCLUSIONS: It is possible to reduce the DUP through an ED program. The reduction in DUP is associated with better clinical status at baseline that is maintained after 3 months. PMID- 14757591 TI - Pharmacotherapy plus psychotherapy for treatment of depression in active injection drug users. AB - CONTEXT: Depressive disorders are common among opiate abusers and are associated with detrimental behavioral effects. However, there is little precedent for offering active drug users complex treatments for depression. OBJECTIVE: To determine whether combined psychotherapy and pharmacotherapy treatment reduces reported depressive symptoms compared with an assessment-only condition among out of-treatment drug injectors. DESIGN: Randomized controlled trial. SETTING: Research office located at an academic medical center. PATIENTS: Active injection drug users with a DSM-IV diagnosis of major depression, dysthymia, substance induced mood disorder with symptoms persisting for at least 3 months, or major depression plus dysthymia, and a Modified Hamilton Rating Scale for Depression (HAM-D) score greater than 13. INTERVENTION: Combined psychotherapy (8 sessions of cognitive behavior therapy) plus pharmacotherapy (citalopram). MAIN OUTCOME MEASURES: Modified HAM-D scale scores at the end of 3 months of combined treatment. RESULTS: The 109 study subjects were 64% male and had a mean age of 36.7 years and a mean baseline HAM-D score of 20.7. Depression subtypes included major depression only (63%), substance-induced depression (17%), and major depression plus dysthymia (17%). In the intent-to-treat analysis, participants in treatment averaged 2.11 HAM-D points greater improvement than control subjects (P=.08), and 26.1% of combined treatment patients (n=53) compared with 12.5% of control patients (n=56) were in remission (P=.047). Nearly 40% of fully adherent subjects (receiving >75% of either psychotherapy or pharmacotherapy) were in remission at follow-up (odds ratio, 3.6; P=.04). CONCLUSIONS: Combined treatment for depression is significantly superior to a control condition (assessment only) in proportion of patients in remission, but not in HAM-D improvement among drug injectors. Full adherence to treatment is associated with the largest treatment effects. Our findings demonstrate that active drug users with dual diagnoses are able to participate in conventional treatment. PMID- 14757592 TI - Increased incidence of diagnosed depressive illness in hypogonadal older men. AB - CONTEXT: Age-associated hypogonadism (testosterone deficit) occurs in 30% of men after the age of 55; it is associated with decreased muscle mass, bone mineral density, and libido, and with anorexia, fatigue, and irritability. Although some of these symptoms overlap with those of depression, the association between the 2 disorders is unclear. OBJECTIVE: To determine if hypogonadal men have an increased incidence of depressive illness compared with eugonadal men. DESIGN: Historical cohort study using computerized medical records, followed by a manual medical record review. SETTING: Veterans Affairs Puget Sound Health Care System. PARTICIPANTS: Two hundred seventy-eight men 45 years and older, without prior diagnosed depressive illness and with consistently normal or low testosterone levels (total testosterone level < or =200 ng/dL [< or =6.94 nmol/L]; or free testosterone level < or =0.9 ng/dL [< or =0.03 nmol/L]) at baseline and during a 2-year follow-up period. MAIN OUTCOME MEASURES: Incidence of, and time to, a depression diagnosis. RESULTS: The 2-year incidence of diagnosed depressive illness was 21.7% in hypogonadal men vs 7.1% in others (chi2(1)=6.0, P=.01). A Kaplan-Meier survival analysis showed a significant difference between hypogonadal and eugonadal men in time to diagnosed depression (log-rank test chi2(1)=6.9, P=.008). We used Cox proportional hazards regression models to examine the association of hypogonadism and time to depression diagnosis, adjusting for age, race, number of clinic visits, alcohol use disorders, prostate cancer, and overall medical comorbidity. The unadjusted hazard ratio for depression with hypogonadism was 3.5 (95% confidence interval, 1.3-9.4) (P=.01). Controlling for all covariates, hypogonadism remained significantly associated with depression (adjusted hazard ratio, 4.2; 95% confidence interval, 1.5-12.0) (P=.008). CONCLUSIONS: Hypogonadal men showed an increased incidence of depressive illness and a shorter time to diagnosis of depression. Further prospective studies are needed to confirm these preliminary findings and to clarify the role of testosterone in the treatment of depressive illness in older men. PMID- 14757593 TI - Regional cerebral blood flow in the amygdala and medial prefrontal cortex during traumatic imagery in male and female Vietnam veterans with PTSD. AB - CONTEXT: Theoretical neuroanatomic models of posttraumatic stress disorder (PTSD) and the results of previous neuroimaging studies of PTSD highlight the potential importance of the amygdala and medial prefrontal regions in this disorder. However, the functional relationship between these brain regions in PTSD has not been directly examined. OBJECTIVE: To examine the relationship between the amygdala and medial prefrontal regions during symptom provocation in male combat veterans (MCVs) and female nurse veterans (FNVs) with PTSD. DESIGN: Case-control study. SETTING: Academic medical center. PARTICIPANTS: Volunteer sample of 17 (7 men and 10 women) Vietnam veterans with PTSD (PTSD group) and 19 (9 men and 10 women) Vietnam veterans without PTSD (control group). MAIN OUTCOME MEASURES: We used positron emission tomography and the script-driven imagery paradigm to study regional cerebral blood flow (rCBF) during the recollection of personal traumatic and neutral events. Psychophysiologic and emotional self-report data also were obtained to confirm the intended effects of script-driven imagery. RESULTS: The PTSD group exhibited rCBF decreases in medial frontal gyrus in the traumatic vs neutral comparison. When this comparison was conducted separately by subgroup, MCVs and FNVs with PTSD exhibited these medial frontal gyrus decreases. Only MCVs exhibited rCBF increases in the left amygdala. However, for both subgroups with PTSD, rCBF changes in medial frontal gyrus were inversely correlated with rCBF changes in the left amygdala and the right amygdala/periamygdaloid cortex. Furthermore, in the traumatic condition, for both subgroups with PTSD, symptom severity was positively related to rCBF in the right amygdala and negatively related to rCBF in medial frontal gyrus. CONCLUSIONS: These results suggest a reciprocal relationship between medial prefrontal cortex and amygdala function in PTSD and opposing associations between activity in these regions and symptom severity consistent with current functional neuroanatomic models of this disorder. PMID- 14757594 TI - Reduced hippocampal volumes associated with the long variant of the serotonin transporter polymorphism in major depression. AB - BACKGROUND: Substantial evidence supports a role for dysfunction of the serotonin transporter in the pathogenesis of major depression. Several studies have found reciprocal interactions between the serotonergic system and both brain-derived neurotrophic factor and glutamate, which are known to modulate or affect hippocampal morphologic characteristics. OBJECTIVE: To examine the influence of a polymorphism (5-HTTLPR) in the promoter region of the serotonin transporter gene on hippocampal volumes in patients with major depression and healthy controls. DESIGN: Baseline investigation of a prospective magnetic resonance imaging study with a 4-year follow-up period. PATIENTS: We examined 40 inpatients with major depression as well as 40 healthy controls matched for age, sex, and handedness. MAIN OUTCOME MEASURES: Subjects underwent high-resolution magnetic resonance imaging. Furthermore, genotyping for the 5-HTTLPR biallelic polymorphism was performed, which consists of a 44-base pair insertion (L allele) or deletion (S allele). RESULTS: Patients with the L/L homozygous genotype had significantly smaller hippocampal gray matter (left hemisphere: P=.003; right hemisphere: P=.01) and white matter volumes (left hemisphere: P=.001; right hemisphere: P=.002) than controls with this genotype. No significant differences were found between patients and controls with the L/S or S/S genotype. Moreover, patients with the L/L genotype had significantly smaller hippocampal white matter volumes than those with the L/S or S/S genotype (P=.03). CONCLUSIONS: These findings suggest that homozygosity for the L allele is associated with decreased hippocampal volumes in patients with major depression but not in healthy controls. A possible explanation is that the interaction between the serotonergic system and neurotrophic factors as well as excitatory amino acid neurotransmission may affect hippocampal morphologic characteristics. PMID- 14757595 TI - A 4-year longitudinal study on risk factors for alcoholism. AB - BACKGROUND: Longitudinal studies are needed to resolve inconsistencies in previous findings regarding antecedents of alcoholism. OBJECTIVE: To investigate genetic and environmental risk factors for alcoholism. DESIGN: A 4-year longitudinal cohort study. SETTING: General community. PARTICIPANTS: A population based cohort was randomly selected from 4 aboriginal groups in Taiwan. Cohort subjects free from any alcohol use disorder at phase 1 (n=499) were reassessed approximately 4 years later (phase 2). The percentage of participants who completed the study was 98.4%. MAIN OUTCOME MEASURES: A standardized semistructured clinical interview for alcoholism and other psychiatric comorbidity was used in both phases of the study. The main outcome measure was the incidence of alcohol use disorder. Specific risk factors examined included sociodemographic factors, family history of alcoholism, extent of acculturation, psychiatric comorbidity, and alcohol-metabolizing genes. RESULTS: Using Cox proportional hazards regression analysis, the risk for alcoholism was significantly higher among subjects who were male (odds ratio [OR], 2.78; 95% confidence interval [CI], 1.79-4.32), aged 15 to 24 years (OR, 5.05; 95% CI, 2.06 6.18), unmarried (OR, 1.60; 95% CI, 1.03-2.49), and employed (OR, 2.25; 95% CI, 1.34-3.77) and had a higher educational level (OR, 1.76; 95% CI, 1.12-2.75), a family history of alcoholism (OR, 1.73; 95% CI, 1.06-2.83), and a higher extent of cultural assimilation (OR, 2.07; 95% CI, 1.28-3.35). Two specific risk pathways emerged on multivariate analysis: the highest risk was among subjects aged 25 to 34 years with anxiety disorders (OR, 16.86; 95% CI, 3.98-71.41), and the other was among men with the less active ADH2*1 gene (OR, 5.87; 95% CI, 2.73 12.60). CONCLUSION: Based on incidence cases of alcoholism among aboriginal Taiwanese, this study confirms the significant roles of anxiety disorders and of the ADH2*1 allele as antecedents of alcoholism among specific age and sex groups. PMID- 14757596 TI - Application of a latent class analysis to empirically define eating disorder phenotypes. AB - CONTEXT: Diagnostic criteria for eating disorders influence how we recognize, research, and treat eating disorders, and empirically valid phenotypes are required for revealing their genetic bases. OBJECTIVE: To empirically define eating disorder phenotypes. DESIGN: Data regarding eating disorder symptoms and features from 1179 individuals with clinically significant eating disorders were submitted to a latent class analysis. The resulting latent classes were compared on non-eating disorder variables in a series of validation analyses. SETTING: Multinational, collaborative study with cases ascertained through diverse clinical settings (inpatient, outpatient, and community). PARTICIPANTS: Members of affected relative pairs recruited for participation in genetic studies of eating disorders in which probands met DSM-IV-TR criteria for anorexia nervosa (AN) or bulimia nervosa and had at least 1 biological relative with a clinically significant eating disorder. Main Outcome Measure Number and clinical characterization of latent classes. RESULTS: A 4-class solution provided the best fit. Latent class 1 (LC1) resembled restricting AN; LC2, AN and bulimia nervosa with the use of multiple methods of purging; LC3, restricting AN without obsessive-compulsive features; and LC4, bulimia nervosa with self-induced vomiting as the sole form of purging. Biological relatives were significantly likely to belong to the same latent class. Across validation analyses, LC2 demonstrated the highest levels of psychological disturbance, and LC3 demonstrated the lowest. CONCLUSIONS: The presence of obsessive-compulsive features differentiates among individuals with restricting AN. Similarly, the combination of low weight and multiple methods of purging distinguishes among individuals with binge eating and purging behaviors. These results support some of the distinctions drawn within the DSM-IV-TR among eating disorder subtypes, while introducing new features to define phenotypes. PMID- 14757597 TI - Does Medicare managed care provide equal treatment for mental illness across races? AB - BACKGROUND: While disparities in access to care are well documented, little is known about the quality of mental health care received by racial and ethnic minorities. We examined the quality of mental health care received by elderly enrollees in Medicare + Choice plans. METHODS: An observational study was performed using individual-level Health Plan Employer Data and Information Set data. From 4182 to 5,016,028 individuals 65 years or older and enrolled in Medicare + Choice plans in 1999 were involved in different measures. Rates of mental health inpatient discharges, average length of stay, percentage of members receiving mental health services, rates of follow-up after hospitalization for mental illness, optimal practitioner contacts for antidepressant medication management, and effective acute- and continuation-phase treatment were assessed. RESULTS: Compared with whites, minorities received substantially less follow-up after hospitalization for mental illness. The 30-day follow-up rates for whites, African Americans, Asians, and Hispanics were 60.2%, 42.4%, 54.1%, and 52.6%, respectively. Minorities also had lower rates of antidepressant medication management for newly diagnosed episodes of depression. The rates of optimal practitioner contacts for whites, African Americans, Asians, and Hispanics were 12.5%, 12.0%, 11.1%, and 10.6%; the rates of effective acute-phase treatment were 60.1%, 48.5%, 40.7%, and 57.6%; and the rates of effective continuation-phase treatment were 46.7%, 32.7%, 31.9%, and 39.6%, respectively. The statistically significant disparities persisted after adjusting for effects of age, sex, income, plan model, profit status, and region of the country. CONCLUSIONS: The overall quality of mental health care for people enrolled in Medicare + Choice managed care plans is far from optimal. There are large and persistent racial differences that merit further attention to better understand their underlying causes and solutions. PMID- 14757599 TI - Can preparedness for biological terrorism save us from pertussis? PMID- 14757600 TI - Growth hormone treatment for idiopathic short stature: implications for practice and policy. PMID- 14757601 TI - Managing bronchiolitis and respiratory syncytial virus: finding the yellow brick road. PMID- 14757602 TI - A randomized trial of nebulized epinephrine vs albuterol in the emergency department treatment of bronchiolitis. AB - OBJECTIVE: To determine if nebulized epinephrine is more efficacious than nebulized albuterol in the emergency department (ED) treatment of moderately ill infants with bronchiolitis. METHODS: Sixty-six patients between 0 and 12 months of age with new-onset wheezing, an antecedent upper respiratory tract infection, and a clinical score (Respiratory Distress Assessment Instrument) of 8 to 15 were randomized in a double-blind fashion to receive either 0.9 mg/kg of nebulized 2.25% racemic epinephrine (n = 34) or 0.15 mg/kg of nebulized 0.5% albuterol sulfate (n = 32) at 0, 30, and 60 minutes. MAIN OUTCOME MEASURES: Primary outcome measures were clinical score and respiratory rate. Secondary outcome measures were room air oxygen saturation, elapsed time to meeting clinical criteria for ED discharge, hospitalization rate, and proportion of patients relapsed within 72 hours of ED discharge (relapse rate). RESULTS: Both treatment groups experienced a similar pattern of change in mean clinical score, respiratory rate, and room air saturation over time. There were no significant differences between the groups by these same measures at any time. The median time at which infants were well enough for ED discharge was 90 minutes in the epinephrine-treated group vs 120 minutes in the albuterol-treated group (P =.01). Sixteen infants (47.1%) in the epinephrine-treated group were hospitalized compared with 12 infants (37.5%) in the albuterol-treated group (relative risk, 1.25; 95% confidence interval, 0.71-2.22). Relapse rate was 18.8% (3/16) in the epinephrine-treated group and 42.1% (8/19) in the albuterol-treated group (relative risk, 0.45; 95% confidence interval, 0.14-1.41). Adverse effects occurred infrequently. CONCLUSIONS: Although the patients treated with epinephrine were judged well enough for ED discharge significantly earlier than the patients treated with albuterol, epinephrine was not found to be more efficacious than albuterol in treating moderately ill infants with bronchiolitis. PMID- 14757603 TI - Diagnosis and testing in bronchiolitis: a systematic review. AB - BACKGROUND: The diagnosis of bronchiolitis is based on typical history and results of a physical examination. The indications for and utility of diagnostic and supportive laboratory testing (eg, chest x-ray films, complete blood cell counts, and respiratory syncytial virus testing) are unclear. OBJECTIVES: To review systematically the data on diagnostic and supportive testing in the management of bronchiolitis and to assess the utility of such testing. DESIGN: In conjunction with an expert panel, we generated admissibility criteria and derived relevant terms to search the literature published from 1980 to November 2002 in MEDLINE and the Cochrane Collaboration Database of Controlled Clinical Trials. Trained abstractors completed detailed data collection forms for each article. We summarized the data in tables after performing data integrity checks. RESULTS: Of the 797 abstracts identified, we present evidence from 82 trials that met our inclusion criteria (17 are primary articles on diagnosis of bronchiolitis and 65are reports of treatment or prevention trials). Numerous studies demonstrate that rapid respiratory syncytial virus tests have acceptable sensitivity and specificity, but no data show that respiratory syncytial virus testing affects clinical outcomes in typical cases of the disease. Seventeen studies presented chest x-ray film data. Abnormalities on chest x-ray films ranged from 20% to 96%. Insufficient data exist to show that chest x-ray films reliably distinguish between viral and bacterial disease or predict severity of disease. Ten studies included complete blood cell counts, but most did not present specific results. In one study, white blood cell counts correlated with radiologically defined disease categories of bronchiolitis. CONCLUSIONS: A large number of studies include diagnostic and supportive testing data. However, these studies do not define clear indications for such testing or the impact of testing on relevant patient outcomes. Given the high prevalence of this disease, prospective studies of the utility of such testing are needed and feasible. PMID- 14757604 TI - Pharmacologic treatment of bronchiolitis in infants and children: a systematic review. AB - BACKGROUND: Bronchiolitis is the most common lower respiratory tract infection in infants. Up to 3% of all children in their first year of life are hospitalized with bronchiolitis. Bronchodilators and corticosteroids are commonly used treatments, but little consensus exists about optimal management strategies. OBJECTIVE: To conduct a systematic review of the effectiveness of commonly used treatments for bronchiolitis in infants and children. DATA SOURCES: We searched MEDLINE and the Cochrane Controlled Trials Register for references to randomized controlled trials of bronchiolitis treatment published since 1980. STUDY SELECTION: Randomized controlled trials of interventions for bronchiolitis in infants and children were included if they were published in English between 1980 and November 2002 and had a minimum sample size of 10. DATA EXTRACTION: We abstracted data on characteristics of the study population, interventions used, and results of studies meeting entry criteria into evidence tables and analyzed them by drug category. DATA SYNTHESIS: Interventions were grouped by drug category and qualitatively synthesized. RESULTS: Of 797 abstracts identified in the literature search, we included 54 randomized controlled trials. This review includes 44 studies of the most common interventions: epinephrine (n = 8), beta2 agonist bronchodilators (n = 13), corticosteroids (n = 13), and ribavirin (n = 10). Studies were, in general, underpowered to detect statistically significant outcome differences between study groups. Few studies collected data on outcomes that are of great importance to parents and clinicians, such as the need for and duration of hospitalization. CONCLUSIONS: Overall, little evidence supports a routine role for any of these drugs in treating patients with bronchiolitis. A sufficiently large, well-designed pragmatic trial of the commonly used interventions for bronchiolitis is needed to determine the most effective treatment strategies for managing this condition. PMID- 14757605 TI - Gun carrying and conduct disorder: a highly combustible combination? Implications for juvenile justice and mental and public health. AB - OBJECTIVES: To examine concealed gun carrying between the ages of 12 and 17 years in a population of clinic-referred boys, many of whom qualified for a disruptive behavior disorder, including conduct disorder (CD); to identify factors and diagnoses related to concealed gun carrying; and to examine the extent to which gun carrying is associated with crime in adulthood. DESIGN: Longitudinal follow up study. SETTING: Pittsburgh, Pa, and Athens and Atlanta, Ga. PARTICIPANTS: One hundred seventy-seven clinic-referred boys, first assessed between the ages of 7 and 12 years and followed up yearly until the age of 19 years. MAIN OUTCOME MEASURES: Violence, property offenses, and drug charges in adulthood. RESULTS: Between the ages of 12 and 17 years, 1 in 5 participants carried a concealed gun, and the annual prevalence increased linearly with age. More than half (61.1%) carried a gun for 1 year only. Gun carrying was significantly (incident rate ratio, 3.93%; 95% confidence interval, 1.60-9.60) associated with CD. Conduct disorder, maternal psychopathy, victimization, and parental monitoring increased the risk of gun carrying by a factor of 8. Adult crime was best predicted by gun carrying, CD, and parental monitoring. Gun carrying predicted drug charges, but not violence or property offenses. CONCLUSIONS: Even though the carrying of handguns by juveniles is prohibited, young men with symptoms of CD are more likely to carry guns than young men without CD. The findings are discussed in terms of the need for the inclusion of gun carrying among the symptoms of CD. PMID- 14757606 TI - Barriers to public health management of a pertussis outbreak in Arkansas. AB - BACKGROUND: During the 2001-2002 respiratory season, Arkansas experienced one of its worst pertussis outbreaks. This crisis occurred shortly after the September 11, 2001, terrorist attacks. OBJECTIVE: To determine whether vulnerabilities in the public health infrastructure existed in the context of emerging infectious diseases or possible bioterrorist attacks. DESIGN: Key personnel involved in the Arkansas pertussis outbreak were interviewed, and health department epidemiologic data were reviewed. SETTING: Observations were made for the statewide private public management of the epidemic. PARTICIPANTS: Physicians, infectious disease specialists, epidemiologists, field nurses, health department staff, laboratory staff, and administrators. MAIN OUTCOME MEASURES: Diagnosis capability; vaccine, prophylaxis, and treatment programs; and effectiveness of global outbreak management. RESULTS: Diagnosis of pertussis was a major barrier to management of the outbreak. The nonspecific clinical diagnosis, unreliability of testing methods, excessive number of samples, unavailability of reagents, and inadequate transport system, laboratory personnel, and equipment all impeded effective diagnosis. Vaccine shortage was not believed to contribute to the extent of the outbreak. Prophylaxis was problematic because of feared adverse effects of drugs and uncertainty about the efficacy of new drugs, but compliance was found to be good. From a public health perspective, isolation procedures, school absence policies, and health department referrals to private physicians all contributed to confusion. Problems with communications, staffing, and public cooperation were identified. Despite these barriers, the epidemic was well tolerated, with no known mortality and limited morbidity. CONCLUSIONS: Despite many identified barriers to effective public health management, Arkansas tolerated its worst epidemic of pertussis in many years. However, were the state to experience an outbreak of a more pathogenic agent, introduced either naturally or of bioterrorist origin, these vulnerabilities could become critical. Natural outbreaks serve as excellent experiences on which to recognize and correct barriers to public health management. PMID- 14757607 TI - Breathing patterns in prepubertal children with sleep-related breathing disorders. AB - OBJECTIVE: To investigate abnormal breathing patterns during sleep in prepubertal children using nonstandard polysomnographic patterns in association with an apnea hypopnea scoring technique. PATIENTS AND METHODS: Study participants included 400 children with suspected sleep-related breathing disorders and 60 control children. We analyzed clinical signs and symptoms at entry into the study and 3 months after otolaryngological treatment. We determined the frequency of predefined breathing patterns during sleep through blind analysis of polysomnograms obtained once in control subjects and twice in children referred to our clinic (before and after adenotonsillectomy), using the nasal cannula pressure transducer system, mouth thermistor, esophageal manometry, microphone, and pulse oximetry. We also determined the relationship between breathing patterns during sleep and residual postsurgery symptoms. Further analysis was performed of symptoms and polysomnographic patterns in those children who underwent new treatment interventions due to persistence of symptoms and abnormal polysomnogram findings. RESULTS: Tachypnea, persistently elevated breathing effort, progressively increased breath effort, and discrete flattening of nasal airflow monitored with the nasal cannula-pressure transducer system without oxygen saturation decreases help determine disorder as much as apneas and hypopneas. Abnormal, nonstandard breathing patterns were associated with the same symptoms as those in children with apnea and hypopnea and were more commonly present when there was incomplete resolution of initial symptoms that led treating practitioners to request further treatment. CONCLUSION: Currently published polysomnographic scoring recommendations overlook common breathing abnormalities during sleep that are associated with clinical complaints. PMID- 14757608 TI - Strategies to improve immunization rates and well-child care in a disadvantaged population: a cluster randomized controlled trial. AB - OBJECTIVE: To measure the effect of a multimodal intervention on well-child care visit (WCV) and immunization rates in an inner-city population. DESIGN: Cluster randomized controlled trial. SETTING AND PARTICIPANTS: One-year cohort of 2843 infants born at a hospital in an integrated inner-city health care system. INTERVENTIONS: Eleven clinics were randomly allocated to 1 of 3 study arms: WCV intervention (n = 3), immunization intervention (n = 4), and controls (n = 4). Interventions to improve immunization and WCV rates included both patient-based and clinic-based activities. MAIN OUTCOME MEASURES: Up-to-date status with childhood immunizations and WCVs by age 12 months (primary) and health care utilization and charges (secondary). RESULTS: Compared with the control arm, the WCV and immunization arms had 5% to 6% higher immunization rates and 7% to 8% higher WCV rates. In multivariate analyses that accounted for the clustered nature of the data, the number of immunizations received was greater in the WCV arm than in controls. However, neither the WCV nor the immunization intervention increased WCV or immunization up-to-date rates. The WCV arm had slightly higher health care charges. Neither intervention affected emergency, urgent care or inpatient utilization. CONCLUSIONS: This multimodal intervention produced a small increase in the number of childhood immunizations delivered. However, patient- and clinic-based methods did not lead to significant increases in WCV or immunization up-to-date rates after controlling for other factors. Methods found in some settings to increase immunization up-to-date rates may not be as effective in a population of inner-city socioeconomically disadvantaged children. PMID- 14757609 TI - An intervention to reduce television viewing by preschool children. AB - BACKGROUND: Television viewing has been associated with increased violence in play and higher rates of obesity. Although there are interventions to reduce television viewing by school-aged children, there are none for younger children. OBJECTIVE: To develop and evaluate an intervention to reduce television viewing by preschool children. DESIGN: Randomized controlled trial conducted in 16 preschool and/or day care centers in rural upstate New York. PATIENTS: Children aged 2.6 through 5.5 years. INTERVENTION: Children attending intervention centers received a 7-session program designed to reduce television viewing as part of a health promotion curriculum, whereas children attending the control centers received a safety and injury prevention program. OUTCOME MEASUREMENTS: Change in parent-reported child television/video viewing and measured growth variables. RESULTS: Before the intervention, the intervention and control groups viewed 11.9 and 14.0 h/wk of television/videos, respectively. Afterward, children in the intervention group decreased their television/video viewing 3.1 h/wk, whereas children in the control group increased their viewing by 1.6 h/wk, for an adjusted difference between the groups of -4.7 h/wk (95% confidence interval, 8.4 to -1.0 h/wk; P =.02). The percentage of children watching television/videos more than 2 h/d also decreased significantly from 33% to 18% among the intervention group, compared with an increase of 41% to 47% among the control group, for a difference of -21.5% (95% confidence interval, -42.5% to -0.5%; P =.046). There were no statistically significant differences in children's growth between groups. CONCLUSIONS: This study is the first to show that a preschool based intervention can lead to reductions in young children's television/video viewing. Further research is needed to determine the long-term effects associated with reductions in young children's television viewing. PMID- 14757610 TI - "Benign" extra-axial fluid in survivors of neonatal intensive care. AB - OBJECTIVES: To identify the prevalence of "benign" extra-axial fluid (BEAF), the risk factors associated with this condition, and the natural history in "graduates" of neonatal intensive care. DESIGN: Cross-sectional study. SETTING: Neonatal follow-up clinic at a tertiary care center. PATIENTS: Seventy-seven infants with a head circumference greater than the 95th percentile by growth percentiles from either the National Center for Health Statistics or the Infant Health and Development Program growth percentile graphs who attended the Neonatal Follow-up Program at The Children's Hospital of Philadelphia between January 1, 1998, and December 31, 2001. MAIN OUTCOME MEASURES: Bronchopulmonary dysplasia, extracorporeal membrane oxygenation; development at 18 to 24 months. RESULTS: There were 26 infants (34%) in the BEAF group, 43 (56%) in the control group without extra-axial fluid, and 8 (10%) in the hydrocephalus group. Compared with the control group, infants with BEAF were more likely to have bronchopulmonary dysplasia or to require use of extracorporeal membrane oxygenation in the immediate neonatal period (risk ratio, 6.1; 95% confidence interval, 1.5-29.8). Measurements of head circumference in the BEAF group showed rapid growth between 3 and 12 months, followed by growth greater than and parallel to the 95th percentile. Head circumference measurements in the control group showed continued growth along the 95th percentile for age. Infants with BEAF were more likely than controls to develop cerebral palsy (risk ratio, 9.9; 95% confidence interval, 1.3 77.9) and to have evidence of developmental delay at adjusted ages 12 and 18 to 24 months. CONCLUSION: The presence of extra-axial fluid in macrocephalic survivors of neonatal intensive care is associated with an increased risk of developmental delay and cerebral palsy compared with control macrocephalic survivors. PMID- 14757611 TI - Primary care access and quality: two sides of the same coin. PMID- 14757612 TI - After-hours telephone triage: reducing the financial burden. PMID- 14757613 TI - A prospective study of aspirin use and the risk for colorectal adenoma. AB - BACKGROUND: Randomized, double-blind, placebo-controlled trials have established that regular aspirin use reduces the risk for recurrent colorectal adenoma. However, the effect of dose and duration of use, particularly in an average-risk population, is not well understood. OBJECTIVE: To examine the influence of dose and duration of aspirin use in the primary prevention of colorectal adenoma. DESIGN: Prospective cohort study. SETTING: Nurses' Health Study. PARTICIPANTS: 27 077 women, 34 to 77 years of age, without a history of adenoma, cancer, inflammatory bowel disease, or familial polyposis, who underwent lower endoscopy between 1980 and 1998. MEASUREMENTS: 1368 cases of confirmed distal colorectal adenoma were diagnosed between 1980 and 1998. Self-reported data on aspirin use were collected from biennial questionnaires. RESULTS: After other risk factors for adenoma were adjusted, women who regularly used aspirin (> or =2 standard aspirin tablets/wk) had a multivariate relative risk for adenoma of 0.75 (95% CI, 0.66 to 0.84) compared with nonregular users. Compared with women who denied any aspirin use, the multivariate relative risks for adenoma were 0.80 (CI, 0.70 to 0.93) for women who used 0.5 to 1.5 standard tablets per week, 0.74 (CI, 0.62 to 0.88) for those who used 2 to 5 tablets per week, 0.72 (CI, 0.61 to 0.85) for those who used 6 to 14 tablets per week, and 0.49 (CI, 0.36 to 0.65) for those who used more than 14 tablets per week (P < 0.001 for trend). Similar dose response relationships were found among regular short-term users (< or =5 years; P < 0.001) and long-term users (>5 years; P < 0.001). In contrast, after adjustments were made for dose, increasing duration of aspirin use did not confer greater risk reduction (P > 0.2). CONCLUSIONS: Regular, short-term use of aspirin is inversely associated with risk for colorectal adenoma. However, the greatest protective effect is evident at substantially higher doses (>14 tablets/wk) than those recommended for the prevention of cardiovascular disease. Before aspirin can be recommended for chemoprevention in the general adult population, these results suggest the need for a more thorough evaluation of the risks and benefits of routine aspirin use at doses not previously considered. PMID- 14757614 TI - The metabolic syndrome and chronic kidney disease in U.S. adults. AB - BACKGROUND: The metabolic syndrome is a common risk factor for cardiovascular disease. OBJECTIVE: To examine the association between the metabolic syndrome and risk for chronic kidney disease and microalbuminuria. DESIGN: Cross-sectional study. SETTING: The Third National Health and Nutrition Examination Survey. PATIENTS: Participants 20 years of age or older were studied in the chronic kidney disease (n = 6217) and microalbuminuria (n = 6125) analyses. MEASUREMENTS: The metabolic syndrome was defined as the presence of 3 or more of the following risk factors: elevated blood pressure, low high-density lipoprotein cholesterol level, high triglyceride level, elevated glucose level, and abdominal obesity. Chronic kidney disease was defined as a glomerular filtration rate less than 60 mL/min per 1.73 m2, and microalbuminuria was defined as a urinary albumin creatinine ratio of 30 to 300 mg/g. RESULTS: The multivariate-adjusted odds ratios of chronic kidney disease and microalbuminuria in participants with the metabolic syndrome compared with participants without the metabolic syndrome were 2.60 (95% CI, 1.68 to 4.03) and 1.89 (CI, 1.34 to 2.67), respectively. Compared with participants with 0 or 1 component of the metabolic syndrome, participants with 2, 3, 4, and 5 components of chronic kidney disease had multivariate adjusted odds ratios of 2.21 (CI, 1.16 to 4.24), 3.38 (CI, 1.48 to 7.69), 4.23 (CI, 2.06 to 8.63), and 5.85 (CI, 3.11 to 11.0), respectively. The corresponding multivariate-adjusted odds ratios of microalbuminuria for participants with 3, 4, and 5 components were 1.62 (CI, 1.10 to 2.38), 2.45 (CI, 1.55 to 3.85), and 3.19 (CI, 1.96 to 5.19), respectively. CONCLUSIONS: These findings suggest that the metabolic syndrome might be an important factor in the cause of chronic kidney disease. PMID- 14757615 TI - Low-molecular-weight heparin compared with intravenous unfractionated heparin for treatment of pulmonary embolism: a meta-analysis of randomized, controlled trials. AB - BACKGROUND: Low-molecular-weight heparin has greatly simplified the management of deep venous thrombosis. However, for patients who present with pulmonary embolism, the role of low-molecular-weight heparin is uncertain and unfractionated heparin remains widely used. PURPOSE: To compare the efficacy and safety of fixed-dose subcutaneous low-molecular-weight heparin with that of dose adjusted intravenous unfractionated heparin to treat acute pulmonary embolism. DATA SOURCES: The MEDLINE, EMBASE, and Cochrane Library databases were searched up to 1 August 2003. Additional data sources were manual searches of abstract proceedings and personal contact with investigators and pharmaceutical companies. STUDY SELECTION: Randomized trials comparing fixed-dose subcutaneous low molecular-weight heparin with dose-adjusted intravenous unfractionated heparin for the treatment of nonmassive symptomatic pulmonary embolism or asymptomatic pulmonary embolism in the context of symptomatic deep venous thrombosis. DATA EXTRACTION: Two reviewers independently selected studies and extracted data on study design; quality; and clinical outcomes, including symptomatic venous thromboembolism, death, and major and minor bleeding. Odds ratios for individual outcomes were calculated for each trial and were pooled by using the Mantel Haenszel method. DATA SYNTHESIS: Fourteen trials involving 2110 patients with pulmonary embolism met the inclusion criteria. Separate outcome data for patients with pulmonary embolism were not available from 2 trials (159 patients), leaving 12 trials for meta-analysis. Compared with unfractionated heparin, low-molecular weight heparin was associated with a non-statistically significant decrease in recurrent symptomatic venous thromboembolism at the end of treatment (1.4% vs. 2.4%; odds ratio, 0.63 [95% CI, 0.33 to 1.18]) and at 3 months (3.0% vs. 4.4%; odds ratio, 0.68 [CI, 0.42 to 1.09]). Similar estimates were obtained for patients who presented with symptomatic pulmonary embolism (1.7% vs. 2.3%; odds ratio, 0.72 [CI, 0.35 to 1.48]) or asymptomatic pulmonary embolism (1.2% vs. 3.2%; odds ratio, 0.53 [CI, 0.15 to 1.88]). For major bleeding complications, the odds ratio favoring low-molecular-weight heparin (1.3% vs. 2.1%; odds ratio, 0.67 [CI, 0.36 to 1.27]) was also not statistically significant. CONCLUSIONS: Fixed dose low-molecular-weight heparin treatment appears to be as effective and safe as dose-adjusted intravenous unfractionated heparin for the initial treatment of nonmassive pulmonary embolism. PMID- 14757617 TI - Sources of variation and bias in studies of diagnostic accuracy: a systematic review. AB - BACKGROUND: Studies of diagnostic accuracy are subject to different sources of bias and variation than studies that evaluate the effectiveness of an intervention. Little is known about the effects of these sources of bias and variation. PURPOSE: To summarize the evidence on factors that can lead to bias or variation in the results of diagnostic accuracy studies. DATA SOURCES: MEDLINE, EMBASE, and BIOSIS, and the methodologic databases of the Centre for Reviews and Dissemination and the Cochrane Collaboration. Methodologic experts in diagnostic tests were contacted. STUDY SELECTION: Studies that investigated the effects of bias and variation on measures of test performance were eligible for inclusion, which was assessed by one reviewer and checked by a second reviewer. Discrepancies were resolved through discussion. DATA EXTRACTION: Data extraction was conducted by one reviewer and checked by a second reviewer. DATA SYNTHESIS: The best-documented effects of bias and variation were found for demographic features, disease prevalence and severity, partial verification bias, clinical review bias, and observer and instrument variation. For other sources, such as distorted selection of participants, absent or inappropriate reference standard, differential verification bias, and review bias, the amount of evidence was limited. Evidence was lacking for other features, including incorporation bias, treatment paradox, arbitrary choice of threshold value, and dropouts. CONCLUSIONS: Many issues in the design and conduct of diagnostic accuracy studies can lead to bias or variation; however, the empirical evidence about the size and effect of these issues is limited. PMID- 14757616 TI - Changes in the use of postmenopausal hormone therapy after the publication of clinical trial results. AB - BACKGROUND: The recent publication of clinical trial results has led to a dramatic shift in the evidence about postmenopausal hormone therapy. OBJECTIVE: To examine whether the publication of clinical trial results, specifically the Heart and Estrogen/progestin Replacement Study (HERS) in 1998 and the Women's Health Initiative (WHI) in 2002, has influenced the use of hormone therapy among postmenopausal women. DESIGN: Observational cohort (1997 to 2003). SETTING: San Francisco Mammography Registry, San Francisco, California. PARTICIPANTS: Postmenopausal women between the ages of 50 and 74 years without a personal history of breast cancer who underwent mammography (151862 mammograms). MEASUREMENTS: Self-reported current use of hormone therapy. RESULTS: Among menopausal women who had mammography, it was estimated that 41% were currently using hormone therapy in 1997. Before the publication of HERS, the use of hormone therapy was increasing at a rate of 1% (95% CI, 0% to 2%) per quarter. After the publication of HERS, use decreased by 1% (CI, -3% to 0%) per quarter. In contrast, the publication of the WHI in 2002 was associated with a more substantial decline in the use of hormone therapy of 18% (CI, -21% to -16%) per quarter. Similar associations were observed for most subgroups of women, including women older than 65 years of age; women with a previous hysterectomy; and women who described their race or ethnicity as white, African American, Latina, Chinese, or Filipina. CONCLUSIONS: The release of the HERS data was temporally associated with a modest decline in the use of hormone therapy. In contrast, the release of the principal findings from the WHI was associated with a more substantial decline in use by postmenopausal women. The reason for the differences in decline may relate to the fact that the WHI results were widely publicized or were more applicable to most postmenopausal women because the WHI study was performed in healthy women. PMID- 14757618 TI - Update in cardiology. PMID- 14757619 TI - Effect of alcohol consumption on diabetes mellitus: a systematic review. AB - BACKGROUND: Both diabetes mellitus and alcohol consumption are prevalent in the United States, yet physicians are poorly informed about how alcohol use affects risk for or management of diabetes. PURPOSE: To conduct a systematic review assessing the effect of alcohol use on the incidence, management, and complications of diabetes mellitus in adults. DATA SOURCES: English-language studies in persons 19 years of age or older that were identified by searching the MEDLINE database from 1966 to the third week of August 2003 and the reference lists of key articles. STUDY SELECTION: Two independent assessors reviewed 974 retrieved citations to identify all experimental, cohort, or case-control studies that assessed the effect of alcohol use on diabetes risk, control, self management, adverse drug events, or complications. DATA EXTRACTION: Two independent reviewers extracted data and evaluated study quality on the basis of established criteria. DATA SYNTHESIS: Thirty-two studies that met inclusion criteria were reviewed. Compared with no alcohol use, moderate consumption (one to 3 drinks/d) is associated with a 33% to 56% lower incidence of diabetes and a 34% to 55% lower incidence of diabetes-related coronary heart disease. Compared with moderate consumption, heavy consumption (>3 drinks/d) may be associated with up to a 43% increased incidence of diabetes. Moderate alcohol consumption does not acutely impair glycemic control in persons with diabetes. CONCLUSIONS: Moderate alcohol consumption is associated with a decreased incidence of diabetes mellitus and a decreased incidence of heart disease in persons with diabetes. Further studies are needed to assess the long-term effects of alcohol consumption on glycemic control and noncardiac complications in persons with diabetes. PMID- 14757621 TI - Aspirin prevention of colorectal cancer: more or less? PMID- 14757620 TI - "Special scrutiny": a targeted form of research protocol review. AB - Research participants require ongoing protection of the kind already established in law and regulation. However, "special scrutiny" for certain types of research is also needed. Three criteria for special scrutiny are 1) research that involves initial experiences of translating new scientific advances into humans, especially when the intervention is novel, irreversible, or both; 2) research with a known or credible risk for significant harm (death or serious disability are the clearest examples) to research participants as a consequence of the experimental intervention and with no potential for offsetting direct medical benefit; or 3) research with a protocol that raises ethical questions about research design or implementation for which there is no consensus. Special scrutiny recognizes that not all research protocols are equally ethically challenging and aims to provide appropriate protection for all research participants. PMID- 14757622 TI - The media matter: a call for straightforward medical reporting. PMID- 14757623 TI - Possession. PMID- 14757624 TI - Adverse events following discharge from the hospital. PMID- 14757625 TI - Adverse events following discharge from the hospital. PMID- 14757626 TI - Adverse events following discharge from the hospital. PMID- 14757627 TI - Adverse events following discharge from the hospital. PMID- 14757628 TI - Reversible tricuspid stenosis secondary to massive ascites in hepatic cirrhosis. PMID- 14757629 TI - Mass media-induced availability bias in the clinical suspicion of West Nile fever. PMID- 14757630 TI - Summaries for patients. Relationships between aspirin dose and colorectal adenomas. PMID- 14757631 TI - Summaries for patients. Association of the metabolic syndrome and chronic kidney disease in U.S. adults. PMID- 14757632 TI - Summaries for patients. Treatment of pulmonary embolism. PMID- 14757633 TI - Summaries for patients. Changes in the use of postmenopausal hormone replacement therapy after the publication of study results. PMID- 14757635 TI - Eponyms and the diagnosis of aortic regurgitation. PMID- 14757634 TI - Summaries for patients. Effects of drinking on the risk for diabetes and its complications. PMID- 14757636 TI - Morphogens, their identification and regulation. AB - During the course of development, cells of many tissues differentiate according to the positional information that is set by the concentration gradients of morphogens. Morphogens are signaling molecules that emanate from a restricted region of a tissue and spread away from their source to form a concentration gradient. As the fate of each cell in the field depends on the concentration of the morphogen signal, the gradient prefigures the pattern of development. In this article, we describe how morphogens and their functions have been identified and analyzed, focusing on model systems that have been extensively studied. PMID- 14757637 TI - Myoblast determination in the somatic and visceral mesoderm depends on Notch signalling as well as on milliways(mili(Alk)) as receptor for Jeb signalling. AB - The visceral muscles of the Drosophila midgut consist of syncytia and arise by fusion of founder and fusion-competent myoblasts, as described for the somatic muscles. A single-step fusion results in the formation of binucleate circular midgut muscles, whereas a multiple-step fusion process produces the longitudinal muscles. A prerequisite for muscle fusion is the establishment of myoblast diversity in the mesoderm prior to the fusion process itself. We provide evidence for a role of Notch signalling during establishment of the different cell types in the visceral mesoderm, demonstrating that the basic mechanism underlying the segregation of somatic muscle founder cells is also conserved during visceral founder cell determination. Searching for genes involved in the determination and differentiation of the different visceral cell types, we identified two independent mutations causing loss of visceral midgut muscles. In both of these mutants visceral muscle founder cells are missing and the visceral mesoderm consists of fusion-competent myoblasts only. Thus, no fusion occurs resulting in a complete disruption of visceral myogenesis. Subsequent characterisation of the mutations revealed that they are novel alleles of jelly belly (jeb) and the Drosophila Alk homologue named milliways (mili(Alk)). We show that the process of founder cell determination in the visceral mesoderm depends on Jeb signalling via the Milliways/Alk receptor. Moreover, we demonstrate that in the somatic mesoderm determination of the opposite cell type, the fusion-competent myoblasts, also depends on Jeb and Alk, revealing different roles for Jeb signalling in specifying myoblast diversity. This novel mechanism uncovers a crosstalk between somatic and visceral mesoderm leading not only to the determination of different cell types but also maintains the separation of mesodermal tissues, the somatic and splanchnic mesoderm. PMID- 14757638 TI - Proximodistal subdivision of Drosophila legs and wings: the elbow-no ocelli gene complex. AB - Appendages are thought to have arisen during evolution as outgrowths from the body wall of primitive bilateria. In Drosophila, subsets of body wall cells are set aside as appendage precursors through the action of secreted signaling proteins that direct localized expression of transcription factors. The Drosophila homeodomain protein Distal-less is expressed in the leg primordia and required for formation of legs, but not wings. The homeodomain protein Nubbin is expressed in the wing primordia and required for formation of wings, but not legs. Given that insect legs and wings have a common developmental and evolutionary origin, we sought to identify genes that underlie the specification of all appendage primordia. We present evidence that the zinc-finger proteins encoded by the elbow and no ocelli genes act in leg and wing primordia to repress body wall-specifying genes and thereby direct appendage formation. PMID- 14757639 TI - The AHR-1 aryl hydrocarbon receptor and its co-factor the AHA-1 aryl hydrocarbon receptor nuclear translocator specify GABAergic neuron cell fate in C. elegans. AB - The aryl hydrocarbon receptors (AHR) are bHLH-PAS domain containing transcription factors. In mammals, they mediate responses to environmental toxins such as 2, 3, 7, 8-tetrachlorodibenzo-p-dioxin (TCDD). Such functions of AHRs require a cofactor, the aryl hydrocarbon receptor nuclear translocator (ARNT), and the cytoplasmic chaperonins HSP90 and XAP2. AHR homologs have been identified throughout the animal kingdom. We report here that the C. elegans orthologs of AHR and ARNT, ahr-1 and aha-1, regulate GABAergic motor neuron fate specification. Four C. elegans neurons known as RMED, RMEV, RMEL and RMER express the neurotransmitter GABA and control head muscle movements. ahr-1 is expressed in RMEL and RMER neurons. Loss of function in ahr-1 causes RMEL and RMER neurons to adopt a RMED/RMEV-like fate, whereas the ectopic expression of ahr-1 in RMED and RMEV neurons can transform them into RMEL/RMER-like neurons. This function of ahr-1 requires aha-1, but not daf-21/hsp90. Our results demonstrate that C. elegans ahr-1 functions as a cell-type specific determinant. This study further supports the notion that the ancestral role of the AHR proteins is in regulating cellular differentiation in animal development. PMID- 14757640 TI - Cleavage and secretion is not required for Four-jointed function in Drosophila patterning. AB - Four-jointed (fj) is required for proximodistal growth and planar polarity in Drosophila tissues. It encodes a predicted type II transmembrane protein with putative signal peptidase sites in its transmembrane domain, and its C terminus is secreted. Fj has therefore been proposed to act as a secreted signalling molecule. We show that Fj protein has a graded distribution in eye and wing imaginal discs, and is largely localised to the Golgi in vivo and in transfected cells. Forms of Fj that are constitutively secreted or anchored in the Golgi were assayed for function in vivo. We find that cleavage and secretion of Fj is not necessary for activity, and that Golgi-anchored Fj has increased activity over wild type. fj has similar phenotypes to those caused by mutations in the cadherin encoding genes fat (ft) and dachsous (ds). We show that fj interacts genetically with ft and ds in planar polarity and proximodistal patterning. We propose that Fj may act in the Golgi to regulate the activity of Ft and Ds. PMID- 14757641 TI - Paraxial mesoderm specifies zebrafish primary motoneuron subtype identity. AB - We provide the first analysis of how a segmentally reiterated pattern of neurons is specified along the anteroposterior axis of the vertebrate spinal cord by investigating how zebrafish primary motoneurons are patterned. Two identified primary motoneuron subtypes, MiP and CaP, occupy distinct locations within the ventral neural tube relative to overlying somites, express different genes and innervate different muscle territories. In all vertebrates examined so far, paraxial mesoderm-derived signals specify distinct motoneuron subpopulations in specific anteroposterior regions of the spinal cord. We show that signals from paraxial mesoderm also control the much finer-grained segmental patterning of zebrafish primary motoneurons. We examined primary motoneuron specification in several zebrafish mutants that have distinct effects on paraxial mesoderm development. Our findings suggest that in the absence of signals from paraxial mesoderm, primary motoneurons have a hybrid identity with respect to gene expression, and that under these conditions the CaP axon trajectory may be dominant. PMID- 14757642 TI - Math1 controls cerebellar granule cell differentiation by regulating multiple components of the Notch signaling pathway. AB - Cerebellar granule cells (CGC) are the most abundant neurons in the mammalian brain, and an important tool for unraveling molecular mechanisms underlying neurogenesis. Math1 is a bHLH transcription activator that is essential for the genesis of CGC. To delineate the effects of Math1 on CGC differentiation, we generated and studied primary cultures of CGC progenitors from Math1/lacZ knockout mice. Rhombic lip precursors appeared properly positioned, expressed CGC specific markers, and maintained Math1 promoter activity in vivo and in vitro, suggesting that Math1 is not essential for the initial stages of specification or survival of CGC. Moreover, the continuous activity of Math1 promoter in the absence of MATH1, indicated that MATH1 was not necessary for the activation of its own expression. After 6, but not 3, days in culture, Math1 promoter activity was downregulated in control cultures, but not in cells from Math1 null mice, thus implying that Math1 participates in a negative regulatory feedback loop that is dependent on increased levels of MATH1 generated through the positive autoregulatory feedback loop. In addition, Math1 null CGC did not differentiate properly in culture, and were unable to extend processes. All Notch signaling pathway receptors and ligands tested were expressed in the rhombic lip at embryonic date 14, with highest levels of Notch2 and Jag1. However, Math1-null rhombic lip cells presented conspicuous downregulation of Notch4 and Dll1. Moreover, of the two transcriptional repressors known to antagonize Math1, Hes5 (but not Hes1) was downregulated in Math1-null rhombic lip tissue and primary cultures, and was shown to bind MATH1, thus revealing a negative regulatory feedback loop. Taken together, our data demonstrate that CGC differentiation, but not specification, depends on Math1, which acts by regulating the level of multiple components of the Notch signaling pathway. PMID- 14757643 TI - CUPULIFORMIS establishes lateral organ boundaries in Antirrhinum. AB - Cupuliformis mutants are defective in shoot apical meristem formation, but cup plants overcome this early barrier to development to reach maturity. CUP encodes a NAC-domain transcription factor, homologous to the Petunia NAM and Arabidopsis CUC proteins. The phenotype of cup mutants differs from those of nam and cuc1 cuc2 in that dramatic organ fusion is observed throughout development. In addition to cotyledon and floral organ fusions, severe lateral organ fusion is found in leaves and inflorescences, and the apical meristem becomes highly fasciated. These features reveal a role for CUP in the establishment of all above ground organ boundaries. Consistent with this function, CUP is expressed at the boundaries of all lateral organs and meristems. It is not currently known how NAC domain genes act to establish organ boundaries. Here, we show that CUP directly interacts with a TCP-domain transcription factor. Members of the TCP-domain family have previously been shown to regulate organ outgrowth. Our results suggest a model for the establishment of organ boundaries based on the localised expression of NAC-domain and TCP-domain factors. PMID- 14757644 TI - A direct role for Fgf but not Wnt in otic placode induction. AB - Induction of the otic placode, which gives rise to all tissues comprising the inner ear, is a fundamental aspect of vertebrate development. A number of studies indicate that fibroblast growth factor (Fgf), especially Fgf3, is necessary and sufficient for otic induction. However, an alternative model proposes that Fgf must cooperate with Wnt8 to induce otic differentiation. Using a genetic approach in zebrafish, we tested the roles of Fgf3, Fgf8 and Wnt8. We demonstrate that localized misexpression of either Fgf3 or Fgf8 is sufficient to induce ectopic otic placodes and vesicles, even in embryos lacking Wnt8. Wnt8 is expressed in the hindbrain around the time of otic induction, but loss of Wnt8 merely delays expression of preotic markers and otic vesicles form eventually. The delay in otic induction correlates closely with delayed expression of fgf3 and fgf8 in the hindbrain. Localized misexpression of Wnt8 is insufficient to induce ectopic otic tissue. By contrast, global misexpression of Wnt8 causes development of supernumerary placodes/vesicles, but this reflects posteriorization of the neural plate and consequent expansion of the hindbrain expression domains of Fgf3 and Fgf8. Embryos that misexpress Wnt8 globally but are depleted for Fgf3 and Fgf8 produce no otic tissue. Finally, cells in the preotic ectoderm express Fgf (but not Wnt) reporter genes. Thus, preotic cells respond directly to Fgf but not Wnt8. We propose that Wnt8 serves to regulate timely expression of Fgf3 and Fgf8 in the hindbrain, and that Fgf from the hindbrain then acts directly on preplacodal cells to induce otic differentiation. PMID- 14757646 TI - Academic medicine: resuscitation in progress. PMID- 14757645 TI - Foxa2 regulates alveolarization and goblet cell hyperplasia. AB - The airways are lined by several distinct epithelial cells that play unique roles in pulmonary homeostasis; however, the mechanisms controlling their differentiation in health and disease are poorly understood. The winged helix transcription factor, FOXA2, is expressed in the foregut endoderm and in subsets of respiratory epithelial cells in the fetal and adult lung. Because targeted mutagenesis of the Foxa2 gene in mice is lethal before formation of the lung, its potential role in lung morphogenesis and homeostasis has not been determined. We selectively deleted Foxa2 in respiratory epithelial cells in the developing mouse lung. Airspace enlargement, goblet cell hyperplasia, increased mucin and neutrophilic infiltration were observed in lungs of the Foxa2-deleted mice. Experimental goblet cell hyperplasia caused by ovalbumin sensitization, interleukin 4 (IL4), IL13 and targeted deletion of the gene encoding surfactant protein C (SP-C), was associated with either absent or decreased expression of Foxa2 in airway epithelial cells. Analysis of lung tissue from patients with a variety of pulmonary diseases revealed a strong inverse correlation between FOXA2 and goblet cell hyperplasia. FOXA2 is required for alveolarization and regulates airway epithelial cell differentiation in the postnatal lung. PMID- 14757650 TI - Drawing policy conclusions from uncontrolled studies. PMID- 14757651 TI - How to improve organ donation rates. PMID- 14757652 TI - How to improve organ donation rates. PMID- 14757654 TI - Surgery in palliative care. PMID- 14757655 TI - Stereotactic radiation for pituitary adenoma. PMID- 14757658 TI - Alberta pharmacists seek authority to prescribe. PMID- 14757657 TI - Face transplants technically possible, but "very hazardous". PMID- 14757659 TI - Talking the talk: new online interpretation system. PMID- 14757660 TI - Fujian flu more severe, but not unusual. PMID- 14757661 TI - Health care spending rises 4.6% in 2003. PMID- 14757663 TI - Sources of variation in provincial drug spending. PMID- 14757665 TI - Do pharmacists' presence on rounding teams reduce preventable adverse drug events in hospital general medical units? PMID- 14757666 TI - COPD death rates: projecting a female trajectory. PMID- 14757667 TI - Use of MRI in the identification and treatment of early ischemic stroke lesions. PMID- 14757668 TI - Oseltamivir (Tamiflu) unsafe in infants under 1 year old. PMID- 14757669 TI - Risks of stillbirth and early neonatal death by day of week. AB - BACKGROUND: Higher risks of stillbirth or early neonatal death, or both, have been reported from several countries for births on weekend days. It is unclear whether such higher risks have persisted in recent years. We investigated weekend associated risks of stillbirth and early neonatal death in most Canadian provinces. METHODS: We studied all 3 239 972 births recorded in Canada, excluding Ontario, between 1985 and 1998. The main outcome measures were the relative risks (RRs) of stillbirth and early neonatal death for infants born on weekends versus weekdays. RESULTS: The proportion of births on weekend days was 24% lower than the proportion on weekdays. Infants born on weekend days had slightly but significantly elevated risks of stillbirth (RR 1.06, 95% confidence interval [CI] 1.02-1.09) and early neonatal death (RR 1.11, 95% CI 1.07-1.16). However, the higher risks disappeared after adjustment for gestational age. INTERPRETATION: The crude risks of stillbirth and early neonatal death remained slightly higher for births on weekend days, but the excesses were much smaller than those reported from other countries. PMID- 14757670 TI - Adverse events among medical patients after discharge from hospital. AB - BACKGROUND: Adverse events (AEs) are adverse outcomes caused by medical care. Several studies have indicated that a substantial number of patients experience AEs before or during hospitalization. However, few data describe AEs after hospital discharge. We determined the incidence, severity, preventability and ameliorability of AEs in patients discharged from the general internal medicine service of a Canadian hospital. METHODS: At a multisite Canadian teaching hospital, we prospectively studied patients who were consecutively discharged home or to a seniors' residence from the general internal medicine service during a 14-week interval in 2002. We used telephone interview and chart review to identify outcomes after discharge. Two physicians independently reviewed each outcome to determine if the patient experienced an AE. The severity, preventability and ameliorability of all AEs were classified. RESULTS: During the study period, outcomes were determined for 328 of the 361 eligible patients, who averaged 71 years of age (interquartile range 54-81 years). After discharge, 76 of the 328 patients experienced at least 1 AE (overall incidence 23%, 95% confidence interval [CI] 19%-28%). The AE severity ranged from symptoms only (68% of the AEs) or symptoms associated with a nonpermanent disability (25%) to permanent disability (3%) or death (3%). The most common AEs were adverse drug events (72%), therapeutic errors (16%) and nosocomial infections (11%). Of the 76 patients, 38 had an AE that was either preventable or ameliorable (overall incidence 12%, 95% CI 9%-16%). INTERPRETATION: Approximately one-quarter of patients in our study had an AE after hospital discharge, and half of the AEs were preventable or ameliorable. PMID- 14757671 TI - Adverse events and patient safety in Canadian health care. PMID- 14757672 TI - Death on the waiting list for cardiac surgery. PMID- 14757673 TI - Flawed analysis, implausible results--move on. PMID- 14757674 TI - Chretien's prescription for medicare: a green poultice in lieu of accountability. PMID- 14757675 TI - Salt and water: a simple approach to hyponatremia. AB - Hyponatremia is common in both inpatients and outpatients. Medications are often the cause of acute or chronic hyponatremia. Measuring the serum osmolality, urine sodium concentration and urine osmolality will help differentiate among the possible causes. Hyponatremia in the physical states of extracellular fluid (ECF) volume contraction and expansion can be easy to diagnose but often proves difficult to manage. In patients with these states or with normal or near-normal ECF volume, the syndrome of inappropriate secretion of antidiuretic hormone is a diagnosis of exclusion, requiring a thorough search for all other possible causes. Hyponatremia should be corrected at a rate similar to that at which it developed. When symptoms are mild, hyponatremia should be managed conservatively, with therapy aimed at removing the offending cause. When symptoms are severe, therapy should be aimed at more aggressive correction of the serum sodium concentration, typically with intravenous therapy in the inpatient setting. PMID- 14757680 TI - Acute heart failure complicating acute coronary syndromes: a deadly intersection. PMID- 14757681 TI - Evidence base for pacemaker mode selection: from physiology to randomized trials. PMID- 14757682 TI - Role of percutaneous septal ablation in hypertrophic obstructive cardiomyopathy. PMID- 14757683 TI - Images in cardiovascular medicine. Traumatic false aneurysm of the left ventricle. PMID- 14757684 TI - Clinical management of metabolic syndrome: report of the American Heart Association/National Heart, Lung, and Blood Institute/American Diabetes Association conference on scientific issues related to management. PMID- 14757685 TI - Images in cardiovascular medicine. Dynamic multislice computed tomography of left ventricular function. PMID- 14757686 TI - Lipoprotein-associated phospholipase A2, high-sensitivity C-reactive protein, and risk for incident coronary heart disease in middle-aged men and women in the Atherosclerosis Risk in Communities (ARIC) study. AB - BACKGROUND: Measuring C-reactive protein (CRP) has been recommended to identify patients at high risk for coronary heart disease (CHD) with low LDL cholesterol (LDL-C). Lipoprotein-associated phospholipase A2 (Lp-PLA2) is a proinflammatory enzyme associated primarily with LDL. METHODS AND RESULTS: In a prospective, case cohort study in 12 819 apparently healthy middle-aged men and women in the Atherosclerosis Risk in Communities study, the relation between Lp-PLA2, CRP, traditional risk factors, and risk for CHD events over a period of approximately 6 years was examined in a proportional hazards model, stratified by LDL-C. Lp PLA2 and CRP levels were higher in the 608 cases than the 740 noncases. Both Lp PLA2 and CRP were associated with incident CHD after adjustment for age, sex, and race with a hazard ratio of 1.78 for the highest tertile of Lp-PLA2 and 2.53 for the highest category of CRP versus the lowest categories. Lp-PLA2 correlated positively with LDL-C (r=0.36) and negatively with HDL-C (r=-0.33) but not with CRP (r=-0.05). In a model adjusted for traditional risk factors including LDL-C, the association of Lp-PLA2 with CHD was attenuated and not statistically significant. For individuals with LDL-C below the median (130 mg/dL), Lp-PLA2 and CRP were both significantly and independently associated with CHD in fully adjusted models. For individuals with LDL-C <130 mg/dL, those with both Lp-PLA2 and CRP levels in the highest tertile were at the greatest risk for a CHD event. CONCLUSIONS: Lp-PLA2 and CRP may be complementary in identifying individuals at high CHD risk who have low LDL-C. PMID- 14757687 TI - Anti-inflammatory and profibrinolytic effect of insulin in acute ST-segment elevation myocardial infarction. AB - BACKGROUND: The clinical benefits of insulin previously observed in acute ST segment-elevation myocardial infarction (STEMI) may be partially explained by an anti-inflammatory effect. We assessed this potential effect of insulin in STEMI patients treated with fibrinolytics. METHODS AND RESULTS: Thirty-two patients receiving reteplase were randomly assigned infusions of either insulin at 2.5 U/h, dextrose, and potassium (GIK) or normal saline and potassium (C) for 48 hours. Plasma concentrations of high-sensitivity C-reactive protein (CRP), serum amyloid A (SAA), plasminogen activator inhibitor-1 (PAI-1), creatine kinase (CK), and CK-MB were measured at baseline and sequentially for 48 hours. Total p47phox protein in mononuclear cells was measured in a subgroup of 13 subjects. Baseline CRP and SAA were significantly increased (2- to 4-fold) at 24 and 48 hours in each group (P<0.01). However, in the insulin group, there was a significant (P<0.05) attenuation of the absolute rise in concentration of CRP and SAA from baseline. The absolute increase of CRP and SAA was reduced by 40% (CRP) and 50% (SAA) at 24 hours and at 48 hours compared with the control group. The absolute increase in PAI-1 from baseline and the percentage increase in p47phox over 48 hours were significantly (P<0.05) lower in the insulin-treated group. CK-MB peaked earlier and tended to be lower in insulin-treated subjects, especially in patients with inferior MI. CONCLUSIONS: Insulin has an anti-inflammatory and profibrinolytic effect in patients with acute MI. These effects may contribute to the clinical benefits of insulin in STEMI. PMID- 14757688 TI - Urinary 8-iso-prostaglandin F2alpha as a risk marker in patients with coronary heart disease: a matched case-control study. AB - BACKGROUND: Oxidative stress is involved in the pathophysiology of atherosclerosis, diabetes mellitus, hypertension, obesity, and cigarette smoking, all of these being risk factors for coronary heart disease (CHD). We tested the hypothesis that risk factors of CHD are associated with abundant systemic oxidative stress. METHODS AND RESULTS: We conducted a case-control study with 93 CHD patients and 93 control subjects frequency-matched by age and sex. Urinary excretion of the F2-isoprostane 8-iso-prostaglandin (PG) F2alpha and its major urinary metabolite, 2,3-dinor-5,6-dihydro-8-iso-PGF2alpha, were measured by gas chromatography-tandem mass spectrometry. Body mass index, systolic blood pressure, and C-reactive protein were elevated in CHD patients (P<0.01). Urinary 8-iso-PGF2alpha and 2,3-dinor-5,6-dihydro-8-iso-PGF2alpha also differed, from 77 (interquartile range, 61-101) to 139 (93-231) pmol/mmol creatinine and from 120 (91-151) to 193 (140-275) pmol/mmol in control subjects and case subjects, respectively (P<0.001). 8-iso-PGF2alpha and its metabolite were highly correlated (Spearman's rho=0.664, P<0.001). HDL cholesterol was diminished in CHD patients (P<0.001). All of these characteristics predicted CHD in univariate analysis. In a multivariate model, the odds ratios were increased only for 8-iso-PGF2alpha (> or =131 pmol/mmol, P<0.001) and C-reactive protein (>3 mg/L, P<0.01), ie, by 30.8 (95% CI, 7.7-124) and 7.2 (1.9-27.6), respectively. 8-iso-PGF2alpha was found to be a novel marker in addition to known risk factors of CHD, ie, diabetes mellitus, hypercholesterolemia, hypertension, and smoking. Urinary excretion of 8 iso-PGF2alpha correlated with the number of risk factors for all subjects (P<0.001 for trend). CONCLUSIONS: 8-iso-PGF2alpha is a sensitive and independent risk marker of CHD. PMID- 14757689 TI - Atrial fibrillation originating from persistent left superior vena cava. AB - BACKGROUND: The left superior vena cava (LSVC) is the embryological precursor of the ligament of Marshall, which has been implicated in the initiation and maintenance of atrial fibrillation (AF). Rarely, the LSVC may persist and has been associated with some organized arrhythmias, though not with AF. We report 5 patients in whom the LSVC was a source of ectopy, initiating AF. METHODS AND RESULTS: In 5 patients (4 men; age, 46+/-11 years) with symptomatic drug refractory AF, ectopy from the LSVC resulting in AF was observed after pulmonary vein isolation. The ectopics were spontaneous in 2 and induced by isoproterenol in the others and preceded P-wave onset by 67+/-13 ms. During multielectrode or electroanatomic mapping, venous potentials were recorded circumferentially at the proximal LSVC near its junction with the coronary sinus (CS), but at the mid-LSVC level, they were recorded only on part of the circumference. The LSVC was electrically connected to the lateral left atrium (LA) and through the CS to the right atrium, with 4.1+/-2.3 CS-LSVC and 1.6+/-0.5 LA-LSVC connections per patient. Catheter ablation in the LSVC targeting these connections resulted in electrical isolation in 4 of the 5 patients without complications. After 15+/-10 months, the 4 patients with successful isolation, including 1 who had successful reablation for LA flutter, remained in sinus rhythm without drugs. CONCLUSIONS: The LSVC can be the arrhythmogenic source of AF with connections to the CS and LA. Ablation of these connections resulted in electrical isolation. PMID- 14757690 TI - Volumetric analysis of in-stent intimal hyperplasia in diabetic patients treated with or without abciximab: results of the Diabetes Abciximab steNT Evaluation (DANTE) randomized trial. AB - BACKGROUND: In diabetic patients in the Evaluation of Platelet IIb/IIIa Inhibitor for Stenting (EPISTENT) trial, abciximab reduced target vessel revascularization by approximately 50% compared with placebo. Whether this is a result of a lower restenosis rate caused by inhibition of intimal hyperplasia remains to be defined. METHODS AND RESULTS: The purpose of this study was to determine whether abciximab at the time of stent implantation would reduce in-stent intimal hyperplasia measured by intravascular ultrasound at 6-month follow-up in type 2 diabetics. Ninety-six diabetic patients (96 lesions) who underwent elective stent implantation for a de novo lesion in a native coronary artery were randomly assigned to receive abciximab or no abciximab. In-stent intimal hyperplasia volume, expressed as percentage of stent volume, did not differ between groups: 41.3+/-21.0% for those treated with abciximab versus 40.5+/-18.3% for those treated without abciximab (P=0.9). There were also no significant differences in angiographic minimal luminal diameter at follow-up (1.74+/-0.69 versus 1.66+/ 0.63 mm; P=0.5), late loss (1.03+/-0.63 versus 1.07+/-0.58 mm; P=0.7), restenosis rate (17.8% versus 22.9%; P=0.5), or cumulative incidence of major adverse cardiac events at 12 months (19.1% versus 20.4%; P=0.9). CONCLUSIONS: Six-month intravascular ultrasound volumetric analysis showed that abciximab, at the time of coronary stent implantation, was not associated with a reduction of in-stent intimal hyperplasia in diabetic patients. PMID- 14757691 TI - Clinical and angiographic predictors of restenosis after stent deployment in diabetic patients. AB - BACKGROUND: Restenosis and consequent adverse cardiac events are increased in diabetics undergoing percutaneous coronary intervention. Use of intracoronary stents may ameliorate such risks; however, factors influencing the likelihood of restenosis after stent deployment in this high-risk patient subgroup are unknown. METHODS AND RESULTS: We retrospectively analyzed all stented diabetic patients in 16 studies of percutaneous coronary intervention, all of which underwent core angiographic analysis at Cardialysis, Rotterdam. Univariate and multivariate analyses, with 37 clinical and angiographic variables, compared those with and without restenosis and predicted restenosis rates calculated through the use of reference charts derived from angiographic data. Within the studies, 418 of 3090 (14%) stented patients with 6-month angiographic follow-up had diabetes. Restenosis (> or =50% diameter stenosis at follow-up) occurred in 550 of 2672 (20.6%) nondiabetic and 130 of 418 (31.1%) diabetic patients (P<0.001). Univariate predictors of restenosis in diabetics were smaller vessel reference diameter (RD) (P<0.001), smaller minimal luminal diameter before stenting (P=0.01), smaller minimal luminal diameter and percent diameter stenosis after stenting (P<0.001, P=0.04), greater stented length of vessel (P<0.001), and reduced body mass index (BMI) (P=0.04). With the use of multivariate analysis, only smaller RD (P=0.003), greater stented length of vessel (P=0.04), and reduced BMI (P=0.04) were associated with restenosis. Reference charts demonstrated an incremental risk of restenosis that appears solely dependent on vessel RD. CONCLUSIONS: Restenosis after stent deployment is significantly increased in diabetic patients. Vessel caliber, stented length of vessel, and lower BMI are predictors of in-stent restenosis in patients with diabetes. Furthermore, vessel caliber affected the predicted risk of restenosis incrementally. PMID- 14757692 TI - Cardiovascular events in diabetic and nondiabetic adults with or without history of myocardial infarction. AB - BACKGROUND: Whether diabetic patients without a history of myocardial infarction (MI) have the same risk of coronary heart disease (CHD) events as nondiabetic patients with a history of MI remains controversial. We compared risks of CHD and stroke events and mortality from cardiovascular disease (CVD) in diabetic and nondiabetic men and women with and without a history of MI. METHODS AND RESULTS: We followed a total of 13 790 African American and white men and women ages 45 to 64 years who participated in the Atherosclerosis Risk in Communities study, beginning in 1987 to 1989. There were 634 fatal CHD or nonfatal MI events, 312 fatal or nonfatal strokes, and 358 deaths from CVD during an average of 9 years of follow-up (125 998 person-years). After adjustment for age, sex, race, Atherosclerosis Risk in Communities field center, and multiple baseline risk factors, patients who had a history of MI without diabetes at baseline had 1.9 times the risk of fatal CHD or nonfatal MI (95% CI, 1.35 to 2.56; P<0.001) compared with diabetic patients without a prior history of MI. The nondiabetic patients with MI also had 1.8 times the risk of CVD mortality compared with diabetic patients without MI (95% CI, 1.22 to 2.72; P=0.003). However, stroke risk was similar between diabetic patients without MI and nondiabetic patients with MI (RR, 1.05; 95% CI, 0.61 to 1.79; P=0.87). We also observed that nondiabetic patients with MI had a carotid artery wall thickness similar to diabetic patients without MI (P=0.77). CONCLUSIONS: Diabetic patients without MI had lower risk of CHD events and mortality from CVD compared with nondiabetic patients with MI, but stroke risk was similar between these 2 groups. PMID- 14757693 TI - Coronary bypass surgery performed off pump does not result in lower in-hospital morbidity than coronary artery bypass grafting performed on pump. AB - BACKGROUND: There is increasing evidence that cardiopulmonary bypass (CPB) may be responsible for the morbidity associated with coronary artery bypass grafting (CABG) surgery. Recent developments in cardiac stabilization devices have made CABG without CPB feasible. However, there is conflicting evidence to date from published trials comparing outcomes between CABG performed with and without CPB, with some trials indicating an advantage to the avoidance of CPB and others showing little benefit. METHODS AND RESULTS: In a single-center randomized trial, 300 patients requiring CABG surgery at a single institution were prospectively randomized to have the procedure performed with CPB (n=150) or on the beating heart (n=150). Exclusion criteria for the trial included emergency procedure, concomitant major cardiac procedures, ejection fraction <30%, and reoperation. In hospital outcomes were analyzed on an intention-to-treat basis. A mean of 3.0+/ 0.9 grafts were performed in the CPB group compared with 2.8+/-0.9 grafts in the beating-heart group (P=0.06). There were no significant differences between the CPB group and the beating-heart group in mortality (0.7% versus 1.3%; P=1.0), transfusion (8.7% versus 9.3%), perioperative myocardial infarction (0.7% versus 2.7%; P=0.37), permanent stroke (0% versus 1.3%; P=0.50), new atrial fibrillation (32% versus 25%; P=0.20), and deep sternal wound infection (0.7% versus 0%; P=1.0). The mean time to extubation was 4 hours, the mean stay in the intensive care unit was 22 hours, and the median length of hospitalization was 5 days in both groups (P=NS). CONCLUSIONS: In contrast to published trials, we were unable to demonstrate any advantage with CABG performed without CPB in terms of patient morbidity. Excellent results can be obtained with either surgical approach. PMID- 14757694 TI - Valsartan restores sarcoplasmic reticulum function with no appreciable effect on resting cardiac function in pacing-induced heart failure. AB - BACKGROUND: Although angiotensin II receptor blockade is considered to be useful for the treatment of human heart failure, little beneficial hemodynamic effect has been shown in some experimental failing hearts. In this study, we assessed the effect of an angiotensin II receptor blocker, valsartan, on sarcoplasmic reticulum (SR) function, defectiveness of which is a major pathogenic mechanism in heart failure. METHODS AND RESULTS: SR vesicles were isolated from dog left ventricular muscle (normal or exposed to 4-week rapid ventricular pacing with or without valsartan). In the untreated and valsartan-treated paced dogs, cardiac function showed similar deterioration (compared with before pacing). However, both the density of beta-receptors and the contractile response to dobutamine were greater in the valsartan-treated paced dogs than in the untreated paced dogs. In untreated paced hearts, the ryanodine receptor was protein kinase A hyperphosphorylated, showed an abnormal Ca2+ leak, and had a decreased amount of ryanodine receptor-bound FKBP12.6. No such phenomena were seen in the valsartan treated paced hearts. Both the SR Ca2+ uptake function and the amount of Ca2+ ATPase were decreased in the untreated failing SR, but both were restored in the valsartan-treated SR. CONCLUSIONS: During the development of pacing-induced heart failure, valsartan preserved the density of beta-receptors and concurrently restored SR function without improving resting cardiac function. PMID- 14757695 TI - Ventricular fibrillation scaling exponent can guide timing of defibrillation and other therapies. AB - BACKGROUND: The scaling exponent (ScE) of the ventricular fibrillation (VF) waveform correlates with duration of VF and predicts defibrillation outcome. We compared 4 therapeutic approaches to the treatment of VF of various durations. METHODS AND RESULTS: Seventy-two swine (19.5 to 25.7 kg) were randomly assigned to 1 of 9 groups (n=8 each). VF was induced and left untreated until the ScE reached 1.10, 1.20, 1.30, or 1.40. Animals were treated with either immediate countershock (IC); 3 minutes of CPR before the first countershock (CPR); CPR for 2 minutes, then drugs given with 3 more minutes of CPR before the first shock (CPR-D); or drugs given at the start of CPR with 3 minutes of CPR before the first shock (Drugs+CPR). Return of spontaneous circulation (ROSC) and 1-hour survival were analyzed with chi2 and Kaplan-Meier survival curves. IC was effective when the ScE was low but had decreasing success as the ScE increased. No animals in the 1.30 or 1.40 groups had ROSC from IC (0 of 16). CPR did not improve first shock outcome in the 1.20 CPR group (3 of 8 ROSC). Kaplan-Meier survival analyses indicated that IC significantly delayed time to ROSC in both the 1.3 (P=0.0006) and the 1.4 (P=0.005) groups. CONCLUSIONS: VF of brief to moderate duration is effectively treated by IC. When VF is prolonged, as indicated by an ScE of 1.3 or greater, IC was not effective and delayed time to ROSC. The ScE can help in choosing the first intervention in the treatment of VF. PMID- 14757696 TI - Increased myocardial dysfunction after ischemia-reperfusion in mice lacking glucose-6-phosphate dehydrogenase. AB - BACKGROUND: Free radical injury contributes to cardiac dysfunction during ischemia-reperfusion. Detoxification of free radicals requires maintenance of reduced glutathione (GSH) by NADPH. The principal mechanism responsible for generating NADPH and maintaining GSH during periods of myocardial ischemia reperfusion remains unknown. Glucose-6-phosphate dehydrogenase (G6PD), the rate limiting enzyme in the pentose phosphate pathway, generates NADPH in a reaction linked to the de novo production of ribose. We therefore hypothesized that G6PD is essential for maintaining GSH levels and protecting the heart during ischemia reperfusion injury. METHODS AND RESULTS: Susceptibility to myocardial ischemia reperfusion injury was determined in Langendorff-perfused hearts isolated from wild-type mice (WT) and mice lacking G6PD (G6PD(def)) (20% of WT myocardial G6PD activity). During global zero-flow ischemia, cardiac function was similar between WT and G6PD(def) hearts. On reperfusion, however, cardiac relaxation and contractile performance were greatly impaired in G6PD(def) myocardium, as demonstrated by elevated end-diastolic pressures and decreased percent recovery of developed pressure relative to WT hearts. Contractile dysfunction in G6PD(def) hearts was associated with depletion of total glutathione stores and impaired generation of GSH from its oxidized form. Increased ischemia-reperfusion injury in G6PD(def) hearts was reversed by treatment with the antioxidant MnTMPyP but unaffected by supplementation of ribose stores. CONCLUSIONS: These results demonstrate that G6PD is an essential myocardial antioxidant enzyme, required for maintaining cellular glutathione levels and protecting against oxidative stress induced cardiac dysfunction during ischemia-reperfusion. PMID- 14757697 TI - Implications of upstream glycoprotein IIb/IIIa inhibition and coronary artery stenting in the invasive management of unstable angina/non-ST-elevation myocardial infarction: a comparison of the Thrombolysis In Myocardial Infarction (TIMI) IIIB trial and the Treat angina with Aggrastat and determine Cost of Therapy with Invasive or Conservative Strategy (TACTICS)-TIMI 18 trial. AB - BACKGROUND: TIMI IIIB and TACTICS-TIMI 18 were 2 trials of an early invasive strategy in unstable angina (UA)/non-ST-elevation myocardial infarction (NSTEMI) that were conducted nearly a decade apart but with virtually identical enrollment criteria and designs, except that upstream glycoprotein IIb/IIIa inhibition was mandated and coronary artery stenting was routinely used in TACTICS-TIMI 18. We sought to examine the effect of these advances on clinical outcomes and the benefits of an early invasive strategy in UA/NSTEMI. METHODS AND RESULTS: Patients were stratified on the basis of their TIMI risk score into low-, intermediate-, and high-risk categories. Within each risk category, the rates of clinical outcomes and the benefit of an early invasive strategy were compared. Compared with patients in TIMI IIIB and adjusting for baseline risk, patients in TACTICS-TIMI 18 had lower rates of death, MI, or rehospitalization for acute coronary syndromes (OR, 0.62; P<0.0001). Across both trials, the benefit of an early invasive strategy was significantly greater with increasing baseline risk: OR, 1.39 in low-risk, 0.80 in intermediate-risk, and 0.57 in high-risk patients (P< or =0.004 for interactions). After adjustment for baseline risk, an early invasive strategy tended toward a more favorable result in TACTICS-TIMI 18 than in TIMI IIIB (OR, 0.79; 95% CI, 0.56 to 1.11). CONCLUSIONS: Advances in the care of patients with UA/NSTEMI, including glycoprotein IIb/IIIa inhibition and stenting, were associated with lower rates of death, MI, and rehospitalization for acute coronary syndromes and a trend toward a greater benefit of an early invasive strategy. PMID- 14757698 TI - Combined blockade of the chemokine receptors CCR1 and CCR5 attenuates chronic rejection. AB - BACKGROUND: Chemokine-chemokine receptor interaction and the subsequent recruitment of T-lymphocytes to the graft are early events in the development of chronic rejection of transplanted hearts or cardiac allograft vasculopathy (CAV). In this study, we sought to determine whether blockade of chemokine receptors CCR1 and CCR5 with Met-RANTES affects the development of CAV in a murine model. METHODS AND RESULTS: B6.CH-2(bm12) strain donor hearts were transplanted heterotopically into wild-type C57BL/6 mice (myosin heavy chain II mismatch). Recipients were treated daily with either Met-RANTES or vehicle starting on postoperative day 4 and were euthanized on postoperative days 24 and 56. We found that Met-RANTES significantly reduced intimal thickening in this model of chronic rejection and that Met-RANTES markedly decreased the infiltration of CD4 and CD8 T lymphocytes and MOMA-2+ monocytes/macrophages into transplanted hearts. Met RANTES also suppressed the ex vivo and in vitro proliferative responses of recipient splenocytes to donor antigens. Finally, Met-RANTES treatment was associated with a marked reduction in RANTES/CCL5 and monocyte chemoattractant protein-1 gene transcript levels in the donor hearts. CONCLUSIONS: Antagonism of the chemokine receptors CCR1 and CCR5 with Met-RANTES attenuates CAV development in vivo by reducing mononuclear cell recruitment to the transplanted heart, proliferative responses to donor antigens, and intragraft RANTES/CCL5 and monocyte chemoattractant protein-1 gene transcript levels. These findings suggest that chemokine receptors CCR1 and CCR5 play significant roles in the development of chronic rejection and may serve as potential therapeutic targets. PMID- 14757699 TI - Murine cytomegalovirus infection increases aortic expression of proatherosclerotic genes. AB - BACKGROUND: The possible etiologic role of infection in cardiovascular disease is still debated. Having previously demonstrated that murine cytomegalovirus (MCMV) infection of apolipoprotein (apo) E-/- mice increases atherosclerotic lesion size, we determined if MCMV infection produces proatherogenic changes in aortic gene expression. Additionally, in cholesterol-fed C57BL/6J mice, we examined the effects of MCMV infection on aortic lesion area. METHODS AND RESULTS: C57BL/6J apoE-/- and wild-type C57BL/6J mice were infected with MCMV. At various time points, aortas were collected and pooled. Total RNA was extracted and hybridized to Affymetrix murine chips or analyzed for specific gene expression using TaqMan reverse transcription-polymerase chain reaction. Data from infected and uninfected mice were compared. A separate group of cholesterol-fed C57BL/6J mice were infected with MCMV, and lesion area in the aortic sinus was assessed using oil red O staining. Acute MCMV infection altered aortic expression of atherogenic genes in young apoE-/- and C57BL/6J mice-specifically, monocyte chemoattractant protein-1, monokine induced by interferon-gamma, and interferon-gamma inducible protein 10. Acute infection in adult 9-month-old apoE-/- mice with well established lesions increased aortic expression of monocyte chemoattractant protein-1. Atherosclerotic lesion area in cholesterol-fed C57BL/6J mice was increased after infection with MCMV. CONCLUSIONS: MCMV infection significantly increases atherosclerotic lesion area and aortic expression of atherogenic genes. These infection-induced effects indicate mechanisms by which cytomegalovirus may contribute to atherosclerotic disease initiation and progression and to the precipitation of clinical events. These results additionally add to data compatible with the concept that infection does play an important role in atherosclerotic disease. PMID- 14757700 TI - Characterization of the mouse cold-menthol receptor TRPM8 and vanilloid receptor type-1 VR1 using a fluorometric imaging plate reader (FLIPR) assay. AB - 1. TRPM8 (CMR1) is a Ca(2+)-permeable channel, which can be activated by low temperatures, menthol, eucalyptol and icilin. It belongs to the transient receptor potential (TRP) family, and therefore is related to vanilloid receptor type-1 (VR1, TRPV1). We tested whether substances which are structurally related to menthol, or which produce a cooling sensation, could activate TRPM8, and compared the responses of TRPM8 and VR1 to these ligands. 2. The effects of 70 odorants and menthol-related substances on recombinant mouse TRPM8 (mTRPM8), expressed in HEK293 cells, were examined using a FLIPR assay. In all, 10 substances (linalool, geraniol, hydroxycitronellal, WS-3, WS-23, FrescolatMGA, FrescolatML, PMD38, CoolactP and Cooling Agent 10) were found to be agonists. 3. The EC(50) values of the agonists defined their relative potencies: icilin (0.2+/ 0.1 microM)>FrescolatML (3.3+/-1.5 microM) > WS-3 (3.7+/-1.7 microM) >(-)menthol (4.1+/-1.3 microM) >frescolatMAG (4.8+/-1.1 microM) > cooling agent 10 (6+/-2.2 microM) >(+)menthol (14.4+/-1.3 microM) > PMD38 (31+/-1.1 microM) > WS-23 (44+/ 7.3 microM) > Coolact P (66+/-20 microM) > geraniol (5.9+/-1.6 mM) > linalool (6.7+/-2.0 mM) > eucalyptol (7.7+/-2.0 mM) > hydroxycitronellal (19.6+/-2.2 mM). 4. Known VR1 antagonists (BCTC, thio-BCTC and capsazepine) were also able to block the response of TRPM8 to menthol (IC(50): 0.8+/-1.0, 3.5+/-1.1 and 18+/-1.1 microM, respectively). 5. The Ca(2+) response of hVR1-transfected HEK293 cells to the endogenous VR1 agonist N-arachidonoyl-dopamine was potentiated by low pH. In contrast, menthol- and icilin-activated TRPM8 currents were suppressed by low pH. 6. In conclusion, in the present study, we identified 10 new agonists and three antagonists of TRPM8. We found that, in contrast to VR1, TRPM8 is inhibited rather than potentiated by protons. PMID- 14757701 TI - Nicotine attenuates beta-amyloid peptide-induced neurotoxicity, free radical and calcium accumulation in hippocampal neuronal cultures. AB - 1. Recent studies indicate that neuronal loss in Alzheimer's disease (AD) is accompanied by the deposition of beta-amyloid protein (A beta) in senile plaques. Nicotine as a major component of cigarette smoke has been suggested to have a protective effect for neurons against A beta neurotoxicity. 2. Our present study demonstrates that nicotine protected cultured hippocampal neurons against the A beta-induced apoptosis. Nicotine effectively inhibits apoptosis in hippocampal cultures caused by A beta(25-35) or A beta(1-40) treatment and increase of caspase activity induced by A beta(25-35) or A beta(1-40). 3. Measurements of cellular oxidation and intracellular free Ca(2+) showed that nicotine suppressed A beta-induced accumulation of free radical and increase of intracellular free Ca(2+). 4. Cholinergic antagonist mecamylamine inhibited nicotine-induced protection against A beta-induced caspase-3 activation and ROS accumulation. 5. The data show that the protection of nicotine is partly via nicotinic receptors. Our results suggest that nicotine may be beneficial in retarding the neurodegenerative diseases such as AD. PMID- 14757704 TI - The role of bradykinin, AT2 and angiotensin 1-7 receptors in the EDRF-dependent vasodilator effect of angiotensin II on the isolated mesenteric vascular bed of the rat. AB - 1. The mechanisms involved in the vasodilator actions of angiotensin II (Ang II) have not yet been completely elucidated. We investigated the potential mechanisms that seem to be involved in the Ang II vasodilator effect using rat isolated mesenteric vascular bed (MVB). 2. Under basal conditions, Ang II does not affect the perfusion pressure of MVB. However, in vessels precontracted with norepinephrine, Ang II induces vasodilation followed by vasoconstriction. Vasoconstrictor, but not the vasodilation of Ang II, is inhibited by AT(1) antagonist (losartan). The vasodilator effect of Ang II was not inhibited by AT(2), angiotensin IV and angiotensin 1-7 receptor antagonists alone (PD 123319, divalinal, A 779, respectively). 3. The vasodilator effect of Ang II is significantly reduced by endothelial removal (deoxycholic acid), but not by indomethacin. Inhibition of NO-synthase by N(G)-nitro-l-arginine methyl ester (l NAME) and guanylyl cyclase by 1H-[1,2,3] oxadiazolo [4,4-a] quinoxalin-1-one (ODQ) reduces the vasodilator effect of Ang II. This effect is also reduced by tetraethylammonium (TEA) or l-NAME, and a combination of l-NAME plus TEA increases the inhibitory effect of the antagonists alone. However, indomethacin does not change the residual vasodilator effect observed in vessels pretreated with l-NAME plus TEA. 4. In vessels precontracted with norepinephrine and depolarized with KCl 25 mm or treated with Ca(2+)-dependent K(+) channel blockers (charybdotoxin plus apamin), the effect of Ang II was significantly reduced. However, this effect is not affected by ATP and voltage-dependent K(+) channel blockers (glybenclamide and 4-aminopyridine). 5. Inhibition of kininase II with captopril significantly potentiates the vasodilator effect of bradykinin (BK) and Ang II in the rat MVB. The inhibitory effect of the B(2) receptor antagonist HOE 140 on the vasodilator effect of Ang II is further enhanced by PD 123319 and/or A 779. 6. The present findings suggest that BK plays an important role in the endothelium-dependent vasodilator effect of Ang II. Probably, the link between Ang II and BK release is modulated by receptors that bind PD 123319 and A 779. PMID- 14757703 TI - Overexpression of beta 1-adrenoceptors in adult rat ventricular myocytes enhances CGP 12177A cardiostimulation: implications for 'putative' beta 4-adrenoceptor pharmacology. AB - 1. CGP 12177A mediates cardiostimulation by activation of the 'putative' beta(4) adrenoceptor; however, it has recently been reported that disruption of the beta(1)-adrenoceptor gene abolishes this effect. We have adenovirally overexpressed beta(1)-adrenoceptors in isolated, cultured adult rat ventricular cardiomyocytes and observed the inotropic potency of isoprenaline and CGP 12177A (in the presence of 1 microm propranolol). 2. Isoprenaline was a full inotropic agonist at rat ventricular myocytes (pD(2) 7.69+/-0.12). CGP 12177A was a nonconventional partial agonist (pD(2) 6.34+/-0.09), increasing inotropy and lusitropy, with an intrinsic activity of 0.34 and antagonised by bupranolol. 3. beta(1)-adrenoceptor overexpression enhanced the inotropic potency of isoprenaline by 11.7-fold (pD(2) 8.76+/-0.14) and CGP 12177A by 5.9-fold (7.11+/ 0.10), respectively. Green fluorescent protein (GFP) overexpression did not alter the potency of isoprenaline or CGP 12177A (pD(2) 7.41+/-0.24 and pD(2) 6.60+/ 0.50, respectively). 4. The cardiostimulant effects of CGP 12177A were enhanced by IBMX (phosphodiesterase inhibitor) and decreased by Rp-cAMPS (cAMP antagonist). CGP 12177A also increased cAMP levels. CGP 12177A but not isoprenaline initiated arrhythmias at lower concentrations following beta(1) adrenoceptor overexpression. 5. (125)I-Cyanopindolol saturation binding in Adv.beta(1) myocytes demonstrated approximately 18-fold increase in beta(1) adrenoceptors. (3)H-CGP 12177A saturation binding, in the presence of propranolol, increased approximately 5-fold following overexpression of beta(1) adrenoceptors. 6. This study demonstrates enhanced cardiostimulation by CGP 12177A (in the presence of propranolol) in rat ventricular myocytes overexpressing beta(1)-adrenoceptors, mediated by a Gs/cAMP signalling pathway. 'Putative' beta(4)-adrenoceptor pharmacology appears to be mediated by activation of a novel affinity state of the beta(1)-adrenoceptor. PMID- 14757702 TI - Cannabinoids and neuroinflammation. AB - Growing evidence suggests that a major physiological function of the cannabinoid signaling system is to modulate neuroinflammation. This review discusses the anti inflammatory properties of cannabinoid compounds at molecular, cellular and whole animal levels, first by examining the evidence for anti-inflammatory effects of cannabinoids obtained using in vivo animal models of clinical neuroinflammatory conditions, specifically rodent models of multiple sclerosis, and second by describing the endogenous cannabinoid (endocannabinoid) system components in immune cells. Our aim is to identify immune functions modulated by cannabinoids that could account for their anti-inflammatory effects in these animal models. PMID- 14757705 TI - Molecular analysis of the subtype-selective inhibition of cloned KATP channels by PNU-37883A. AB - 1. In this study, we have used Kir6.1/Kir6.2 chimeric proteins and current recordings to investigate the molecular basis of PNU-37883A inhibition of cloned K(ATP) channels. 2. Rat Kir6.1, Kir6.2 and Kir6.1/Kir6.2 chimeras were co expressed with either SUR2B or SUR1, following RNA injection into Xenopus oocytes, and fractional inhibition of K(ATP) currents by 10 microm PNU-37883A reported. 3. Channels containing Kir6.1/SUR2B were more sensitive to inhibition by PNU-37883A than those containing Kir6.2/SUR2B (mean fractional inhibition: 0.70, cf. 0.07). 4. On expression with SUR2B, a chimeric channel with the Kir6.1 pore and the Kir6.2 amino- and carboxy-terminal domains was PNU-37883A insensitive (0.06). A chimera with the Kir6.1 carboxy-terminus and Kir6.2 amino terminus and pore was inhibited (0.48). These results, and those obtained with other chimeras, suggest that the C-terminus is an important determinant of PNU 37883A inhibition of Kir6.1. Similar results were seen when constructs were co expressed with SUR1. Further chimeric constructs localised PNU-37883A sensitivity to an 81 amino-acid residue section in the Kir6.1 carboxy-terminus. 5. Our data show that structural differences between Kir6.1 and Kir6.2 are important in determining sensitivity to PNU-37883A. This compound may prove useful in probing the structural and functional differences between the two channel subtypes. PMID- 14757706 TI - Sustained clinical efficacy of sulfadoxine-pyrimethamine for uncomplicated falciparum malaria in Malawi after 10 years as first line treatment: five year prospective study. AB - OBJECTIVE: To measure the efficacy of sulfadoxine-pyrimethamine treatment of falciparum malaria in Malawi from 1998 to 2002, after a change from chloroquine to sulfadoxine-pyrimethamine as first line treatment in that country in 1993. DESIGN: Prospective open label drug efficacy study. SETTING: Health centre in large peri-urban township adjacent to Blantyre, Malawi. PARTICIPANTS: People presenting to a health centre with uncomplicated Plasmodium falciparum malaria. MAIN OUTCOME MEASURES: Therapeutic efficacy and parasitological resistance to standard sulfadoxine-pyrimethamine treatment at 14 days and 28 days of follow up. RESULTS: Therapeutic efficacy remained stable, with adequate clinical response rates of 80% or higher throughout the five years of the study. Analysis of follow up to 28 days showed modest but significant trends towards diminishing clinical and parasitological efficacy over time within the study period. CONCLUSION: Contrary to expectations, sulfadoxine-pyrimethamine has retained good efficacy after 10 years as the first line antimalarial drug in Malawi. African countries with very low chloroquine efficacy, high sulfadoxine-pyrimethamine efficacy, and no other immediately available alternatives may benefit from interim use of sulfadoxine-pyrimethamine while awaiting implementation of combination antimalarial treatments. PMID- 14757707 TI - Environmental pollution and the global burden of disease. AB - Exposures to environmental pollution remain a major source of health risk throughout the world, though risks are generally higher in developing countries, where poverty, lack of investment in modern technology and weak environmental legislation combine to cause high pollution levels. Associations between environmental pollution and health outcome are, however, complex and often poorly characterized. Levels of exposure, for example, are often uncertain or unknown as a result of the lack of detailed monitoring and inevitable variations within any population group. Exposures may occur via a range of pathways and exposure processes. Individual pollutants may be implicated in a wide range of health effects, whereas few diseases are directly attributable to single pollutants. Long latency times, the effects of cumulative exposures, and multiple exposures to different pollutants which might act synergistically all create difficulties in unravelling associations between environmental pollution and health. Nevertheless, in recent years, several attempts have been made to assess the global burden of disease as a result of environmental pollution, either in terms of mortality or disability-adjusted life years (DALYs). About 8-9% of the total disease burden may be attributed to pollution, but considerably more in developing countries. Unsafe water, poor sanitation and poor hygiene are seen to be the major sources of exposure, along with indoor air pollution. PMID- 14757708 TI - The impact of environmental pollution on congenital anomalies. AB - Major congenital anomalies are diagnosed in 2-4% of births. In this paper we review epidemiological studies that have specifically looked at congenital anomalies as a possible outcome of community exposure to chemical exposures associated with environmental pollution. These include studies of drinking water contaminants (heavy metals and nitrates, chlorinated and aromatic solvents, and chlorination by-products), residence near waste disposal sites and contaminated land, pesticide exposure in agricultural areas, air pollution and industrial pollution sources, food contamination, and disasters involving accidental, negligent or deliberate chemical releases of great magnitude. We conclude that there are relatively few environmental pollution exposures for which we can draw strong conclusions about the potential to cause congenital anomalies and, if so, the chemical constituents implicated, to provide an evidence base for public health and clinical practice. A precautionary approach should be adopted at both community and individual level. In order to prevent congenital anomalies, one must reduce exposure to potential teratogens before pregnancy is recognized (i.e. preconceptionally and in the first few weeks of pregnancy). It is a challenge to develop effective strategies for preconceptional care within the primary care framework. Prenatal service providers and counsellors need to be aware of the uncertainties regarding environmental pollution when addressing parental concerns. PMID- 14757709 TI - Infertility and environmental pollutants. AB - While it has long been known that female fertility is impaired by oestrogen exposure, it is unclear whether environmental pollutants with weak oestrogenic effects are sufficiently potent and prevalent to have biological effects in humans. Male fertility, or sperm concentration at least, appears to have deteriorated, and there is substantial spatial variation at both national and global level, as well as a genetic component. Sperm morphology and motility are implicated too. There is good evidence for an increase in testicular cancer, and possibly in other conditions that certain spatial characteristics plus evidence on heritability suggest are linked to impaired spermatogenesis. A candidate agent would need to have started increasing in the early 20th century. Weak environmental oestrogens are not responsible. Candidates include agents affecting endogenous maternal oestrogen levels, environmental anti-androgens (although these cannot explain the epidemiological findings), and dioxin and related compounds. Genetic damage should be considered as a unifying hypothesis, possibly focused on the Y-chromosome. PMID- 14757710 TI - Contribution of environmental factors to cancer risk. AB - Environmental carcinogens, in a strict sense, include outdoor and indoor air pollutants, as well as soil and drinking water contaminants. An increased risk of mesothelioma has consistently been detected among individuals experiencing residential exposure to asbestos, whereas results for lung cancer are less consistent. At least 14 good-quality studies have investigated lung cancer risk from outdoor air pollution based on measurement of specific agents. Their results tend to show an increased risk in the categories at highest exposure, with relative risks in the range 1.5-2.0, which is not attributable to confounders. Results for other cancers are sparse. A causal association has been established between exposure to environmental tobacco smoke and lung cancer, with a relative risk in the order of 1.2. Radon is another carcinogen present in indoor air which may be responsible for 1% of all lung cancers. In several Asian populations, an increased risk of lung cancer is present in women from indoor pollution from cooking and heating. There is strong evidence of an increased risk of bladder, skin and lung cancers following consumption of water with high arsenic contamination; results for other drinking water contaminants, including chlorination by-products, are inconclusive. A precise quantification of the burden of human cancer attributable to environmental exposure is problematic. However, despite the relatively small relative risks of cancer following exposure to environmental carcinogens, the number of cases that might be caused, assuming a causal relationship, is relatively large, as a result of the high prevalence of exposure. PMID- 14757711 TI - Air pollution and infection in respiratory illness. AB - The detrimental effects of air pollution on health have been recognized for most of the last century. Effective legislation has led to a change in the nature of the air pollutants in outdoor air in developed countries, while combustion of raw fuels in the indoor environment remains a major health hazard in developing countries. The mechanisms of how these pollutants exert their effects are likely to be different, but there is emerging evidence that the toxic effects of new photochemical pollutants such as nitrogen dioxide are likely to be related to infection. This review discusses the relationship between air pollution and infection and will explore some of the mechanisms of how both could act synergistically to cause respiratory illnesses especially in exacerbating symptoms in individuals with pre-existing respiratory conditions such as asthma and chronic obstructive pulmonary disease. PMID- 14757712 TI - Evaluating evidence on environmental health risks. AB - The assessment of adverse health effects from environmental hazards involves integration of evidence from a variety of sources, including experimental studies, both in animals and humans, in vitro studies, and epidemiological research. It requires an understanding of the sources, nature and levels of exposure to which humans may be subjected, the nature of the health outcome or toxic effect and the mechanisms by which this might occur, the relationship between dose and response, and a knowledge of the variability and susceptibility of potentially exposed populations. After outlining the process of risk assessment, this paper gives an overview of the most relevant human study methods used to investigate environment and health effects and discusses issues such as confounding and effect modification, that are important to consider when interpreting the results from such studies. Future challenges are outlined, such as increasing responsibility required by scientists to the sensitive issue of data protection and confidentiality, and also new opportunities, such as the increasing availability of computerized data, the incorporation of molecular epidemiological methods to aid the investigation of mechanistic pathways and gene environment interactions, and the development and utilization of sophisticated statistical approaches. PMID- 14757713 TI - Environmental effects and skin disease. AB - The skin is the largest organ in the body and one of its main functions is to protect the body from noxious substances, whether they are ultraviolet radiation, toxic chemicals or prolonged/repeated exposure to water. It is the level of exposure that determines if damage to the organism will result. The harm that can occur to the skin with sufficient exposure will be considered. Contact dermatitis, halogen acne, chemical depigmentation, connective tissue diseases and skin cancer are the conditions that will be covered in this chapter, as environmental exposure is important in their aetiologies. Systemic absorption will not be dealt with. Most environmental exposure to harmful substances will occur at work, but exposure may occur at home or during normal day-to-day activities. PMID- 14757714 TI - Ambient air pollution and health. AB - The adverse health effects of air pollution became widely acknowledged after severe pollution episodes occurred in Europe and North America before the 1960s. In these areas, pollutant levels have decreased. During the last 15 years, however, consistent results, mainly from epidemiological studies, have provided evidence that current air pollutant levels have been associated with adverse long and short-term health effects, including an increase in mortality. These effects have been better studied for ambient particle concentrations but there is also substantial evidence concerning gaseous pollutants such as ozone, NO(2) and CO. Attempts to estimate the impact of air pollution effects on health in terms of the attributable number of events indicate that the ubiquitous nature of the exposure results in a considerable public health burden from relatively weak relative risks. PMID- 14757715 TI - Electromagnetic radiation. AB - Electromagnetic fields (EMF) are ubiquitous in modern society. It is well known that exposure to strong fields can result in acute effects, such as burns; the mechanisms behind such effects are well established. There is, however, also a concern that long-term exposure to weak fields might have health effects due to an as-yet unknown mechanism. Because of the already widespread exposure, even small health effects could have profound public health implications. Comprehensive research efforts are therefore warranted, and are indeed ongoing. The strongest evidence for health risks is from exposure to fields generated in connection with use of electric power. As for fields used by telecommunications technology, there is still considerably fewer data available and for the time being there is only very weak support for the existence of health effects. However, extensive research activities are ongoing and much more data will be available in the near future. This situation of scientific uncertainty and considerable public concern creates dilemmas for decision makers. PMID- 14757716 TI - Hazards of heavy metal contamination. AB - The main threats to human health from heavy metals are associated with exposure to lead, cadmium, mercury and arsenic. These metals have been extensively studied and their effects on human health regularly reviewed by international bodies such as the WHO. Heavy metals have been used by humans for thousands of years. Although several adverse health effects of heavy metals have been known for a long time, exposure to heavy metals continues, and is even increasing in some parts of the world, in particular in less developed countries, though emissions have declined in most developed countries over the last 100 years. Cadmium compounds are currently mainly used in re-chargeable nickel-cadmium batteries. Cadmium emissions have increased dramatically during the 20th century, one reason being that cadmium-containing products are rarely re-cycled, but often dumped together with household waste. Cigarette smoking is a major source of cadmium exposure. In non-smokers, food is the most important source of cadmium exposure. Recent data indicate that adverse health effects of cadmium exposure may occur at lower exposure levels than previously anticipated, primarily in the form of kidney damage but possibly also bone effects and fractures. Many individuals in Europe already exceed these exposure levels and the margin is very narrow for large groups. Therefore, measures should be taken to reduce cadmium exposure in the general population in order to minimize the risk of adverse health effects. The general population is primarily exposed to mercury via food, fish being a major source of methyl mercury exposure, and dental amalgam. The general population does not face a significant health risk from methyl mercury, although certain groups with high fish consumption may attain blood levels associated with a low risk of neurological damage to adults. Since there is a risk to the fetus in particular, pregnant women should avoid a high intake of certain fish, such as shark, swordfish and tuna; fish (such as pike, walleye and bass) taken from polluted fresh waters should especially be avoided. There has been a debate on the safety of dental amalgams and claims have been made that mercury from amalgam may cause a variety of diseases. However, there are no studies so far that have been able to show any associations between amalgam fillings and ill health. The general population is exposed to lead from air and food in roughly equal proportions. During the last century, lead emissions to ambient air have caused considerable pollution, mainly due to lead emissions from petrol. Children are particularly susceptible to lead exposure due to high gastrointestinal uptake and the permeable blood-brain barrier. Blood levels in children should be reduced below the levels so far considered acceptable, recent data indicating that there may be neurotoxic effects of lead at lower levels of exposure than previously anticipated. Although lead in petrol has dramatically decreased over the last decades, thereby reducing environmental exposure, phasing out any remaining uses of lead additives in motor fuels should be encouraged. The use of lead-based paints should be abandoned, and lead should not be used in food containers. In particular, the public should be aware of glazed food containers, which may leach lead into food. Exposure to arsenic is mainly via intake of food and drinking water, food being the most important source in most populations. Long-term exposure to arsenic in drinking-water is mainly related to increased risks of skin cancer, but also some other cancers, as well as other skin lesions such as hyperkeratosis and pigmentation changes. Occupational exposure to arsenic, primarily by inhalation, is causally associated with lung cancer. Clear exposure response relationships and high risks have been observed. PMID- 14757717 TI - Health hazards and waste management. AB - Different methods of waste management emit a large number of substances, most in small quantities and at extremely low levels. Raised incidence of low birth weight births has been related to residence near landfill sites, as has the occurrence of various congenital malformations. There is little evidence for an association with reproductive or developmental effects with proximity to incinerators. Studies of cancer incidence and mortality in populations around landfill sites or incinerators have been equivocal, with varying results for different cancer sites. Many of these studies lack good individual exposure information and data on potential confounders, such as socio-economic status. The inherent latency of diseases and migration of populations are often ignored. Waste management workers have been shown to have increased incidence of accidents and musculoskeletal problems. The health impacts of new waste management technologies and the increasing use of recycling and composting will require assessment and monitoring. PMID- 14757718 TI - Contaminants in drinking water. AB - An adequate supply of safe drinking water is one of the major prerequisites for a healthy life, but waterborne disease is still a major cause of death in many parts of the world, particularly in children, and it is also a significant economic constraint in many subsistence economies. The basis on which drinking water safety is judged is national standards or international guidelines. The most important of these are the WHO Guidelines for Drinking Water Quality. The quality of drinking water and possible associated health risks vary throughout the world with some regions showing, for example, high levels of arsenic, fluoride or contamination of drinking water by pathogens, whereas elsewhere these are very low and no problem. Marked variations also occur on a more local level within countries due, for example, to agricultural and industrial activities. These and others are discussed in this chapter. PMID- 14757719 TI - Indoor air pollution: a global health concern. AB - Indoor air pollution is ubiquitous, and takes many forms, ranging from smoke emitted from solid fuel combustion, especially in households in developing countries, to complex mixtures of volatile and semi-volatile organic compounds present in modern buildings. This paper reviews sources of, and health risks associated with, various indoor chemical pollutants, from a historical and global perspective. Health effects are presented for individual compounds or pollutant mixtures based on real-world exposure situations. Health risks from indoor air pollution are likely to be greatest in cities in developing countries, especially where risks associated with solid fuel combustion coincide with risk associated with modern buildings. Everyday exposure to multiple chemicals, most of which are present indoors, may contribute to increasing prevalence of asthma, autism, childhood cancer, medically unexplained symptoms, and perhaps other illnesses. Given that tobacco consumption and synthetic chemical usage will not be declining at least in the near future, concerns about indoor air pollution may be expected to remain. PMID- 14757720 TI - Asthma: environmental and occupational factors. AB - Asthma is in several ways a difficult disease to study. Generally arising in childhood, its pattern is often one of remission and relapse; at any point there are difficulties in translating its characteristic, clinical features into an operational definition. Geographical and temporal patterns in its distribution - whereby the disease appears to have increased in frequency in more 'westernised' countries -suggest strong environmental determinants in its causation although there are, too, undoubted and important genetic influences on both its incidence and presentation. Recent aetiological research has concentrated on the function of allergen exposure or on the role of early-life microbial contact that may regulate the development of a range of childhood allergies, including asthma. To date the 'hygiene hypothesis' offers the most efficient explanation for the distribution of the disease in time and place although convincing evidence for it remains elusive. PMID- 14757721 TI - Noise pollution: non-auditory effects on health. AB - Noise is a prominent feature of the environment including noise from transport, industry and neighbours. Exposure to transport noise disturbs sleep in the laboratory, but not generally in field studies where adaptation occurs. Noise interferes in complex task performance, modifies social behaviour and causes annoyance. Studies of occupational and environmental noise exposure suggest an association with hypertension, whereas community studies show only weak relationships between noise and cardiovascular disease. Aircraft and road traffic noise exposure are associated with psychological symptoms but not with clinically defined psychiatric disorder. In both industrial studies and community studies, noise exposure is related to raised catecholamine secretion. In children, chronic aircraft noise exposure impairs reading comprehension and long-term memory and may be associated with raised blood pressure. Further research is needed examining coping strategies and the possible health consequences of adaptation to noise. PMID- 14757722 TI - Risks associated with ionizing radiation. AB - This paper reviews current knowledge on the deterministic and stochastic risks (the latter including the risk of cancer and of hereditary disease) associated with exposure to ionizing radiation. Particular attention is paid to cancer risks following exposure to man-made low linear energy transfer radiation. Excess cancer risks have been observed in the Japanese atomic bomb survivors and in many medically and occupationally exposed groups. In general, the relative risks among Japanese survivors of atomic-bomb explosions are greater than those among comparable subsets in studies of medically exposed individuals. Cell sterilization largely accounts for the discrepancy in relative risks between these two populations, although other factors may contribute, such as the generally higher underlying cancer risks in the medical series than in the Japanese atomic bomb survivors. Risks among occupationally exposed groups such as nuclear workforces and underground miners are generally consistent with those observed in the Japanese atomic bomb survivors. PMID- 14757724 TI - Genetic differentiation among recently diverged delphinid taxa determined using AFLP markers. AB - In the mid-1990s, a new common dolphin species (Delphinus capensis) was defined in the northeast Pacific using morphological characters and mitochondrial DNA (mtDNA) sequences. This species is sympatric with a second species, Delphinus delphis; morphological differences between the two are slight and it is clear they are closely related. Does the phenotypic distinction result from only a few important genes or from large differences between their nuclear genomes? We used amplified fragment length polymorphism (AFLP) markers to broadly survey the nuclear genomes of these two species to examine the levels of nuclear divergence and genetic diversity between them. Furthermore, to create an evolutionary context in which to compare the level of interspecific divergence found between the two Delphinus taxa, we also examined two distinct morphotypes of the bottlenose dolphin (Tursiops truncatus). A nonmetric multidimensional scaling analysis clearly differentiated both Delphinus species, indicating that significant nuclear genetic differentiation has arisen between the species despite their morphological similarity. However, the AFLP data indicated that the two T. truncatus morphotypes exhibit greater divergence than D. capensis and D. delphis, suggesting that they too should be considered different species. PMID- 14757725 TI - Microsatellite analysis of genetic diversity of the Vietnamese sika deer (Cervus nippon pseudaxis). AB - The Vietnamese sika deer (Cervus nippon pseudaxis) is an endangered subspecies of economic and traditional value in Vietnam. Most living individuals are held in traditional farms in central Vietnam, others being found in zoos around the world. Here we study the neutral genetic diversity and population structure of this subspecies using nine microsatellite loci in order to evaluate the consequences of the limited number of individuals from which this population was initiated and of the breeding practices (i.e., possible inbreeding). Two hundred individuals were sampled from several villages. Our data show both evidence for limited local inbreeding and isolation by distance with a mean F(ST) value of 0.02 between villages. This suggests that exchange of animals occurs at a local scale, at a rate such that highly inbred mating is avoided. However, the genetic diversity, with an expected heterozygosity (H(e)) of 0.60 and mean number of alleles (k) of 5.7, was not significantly larger than that estimated from zoo populations of much smaller census size (17 animals sampled; H(e) = 0.65, k = 4.11). Our results also suggest that the Vietnamese population might have experienced a slight bottleneck. However, this population is sufficiently variable to constitute a source of individuals for reintroduction in the wild in Vietnam. PMID- 14757726 TI - Phylogenetic and phylogeographic analysis of Iberian lynx populations. AB - The Iberian lynx (Lynx pardinus), one of the world's most endangered cat species, is vulnerable due to habitat loss, increased fragmentation of populations, and precipitous demographic reductions. An understanding of Iberian lynx evolutionary history is necessary to develop rational management plans for the species. Our objectives were to assess Iberian lynx genetic diversity at three evolutionary timescales. First we analyzed mitochondrial DNA (mtDNA) sequence variation to position the Iberian lynx relative to other species of the genus LYNX: We then assessed the pattern of mtDNA variation of isolated populations across the Iberian Peninsula. Finally we estimated levels of gene flow between two of the most important remaining lynx populations (Donana National Park and the Sierra Morena Mountains) and characterized the extent of microsatellite locus variation in these populations. Phylogenetic analyses of 1613 bp of mtDNA sequence variation supports the hypothesis that the Iberian lynx, Eurasian lynx, and Canadian lynx diverged within a short time period around 1.53-1.68 million years ago, and that the Iberian lynx and Eurasian lynx are sister taxa. Relative to most other felid species, genetic variation in mtDNA genes and nuclear microsatellites were reduced in Iberian lynx, suggesting that they experienced a fairly severe demographic bottleneck. In addition, the effects of more recent reductions in gene flow and population size are being manifested in local patterns of molecular genetic variation. These data, combined with recent studies modeling the viability of Iberian lynx populations, should provide greater urgency for the development and implementation of rational in situ and ex situ conservation plans. PMID- 14757727 TI - A microsatellite map of the African human malaria vector Anopheles funestus. AB - Microsatellite markers and chromosomal inversion polymorphisms are useful genetic markers for determining population structure in Anopheline mosquitoes. In Anopheles funestus (2N = 6), only chromosome arms 2R, 3R, and 3L are known to carry polymorphic inversions. The physical location of microsatellite markers with respect to polymorphic inversions is potentially important information for interpreting population genetic structure, yet none of the available marker sets have been physically mapped in this species. Accordingly, we mapped 32 polymorphic A. funestus microsatellite markers to the polytene chromosomes using fluorescent in situ hybridization (FISH) and identified 16 markers outside of known polymorphic inversions. Here we provide an integrated polytene chromosome map for A. funestus that includes the breakpoints of all known polymorphic inversions as well as the physical locations of microsatellite loci developed to date. Based on this map, we suggest a standard set of 16 polymorphic microsatellite markers that are distributed evenly across the chromosome complement, occur predominantly outside of inversions, and amplify reliably. Adoption of this set by researchers working in different regions of Africa will facilitate metapopulation analyses of this primary malaria vector. PMID- 14757728 TI - Identification of quantitative trait loci affecting sex determination in the Eastern treehole mosquito (Ochlerotatus triseriatus). AB - Laboratory colonies of the eastern treehole mosquito (Ochlerotatus triseriatus (Say)) exhibit a consistent female-biased sex ratio. This is unusual among mosquito species, in which heritable sex ratio distortion is usually male biased and mediated by meiotic drive. Quantitative trait loci (QTL) affecting sex were mapped in an F(1) intercross to better understand the genetics underlying this female bias. In P(1) and F(1) parents and in 146 F(2) individuals with a female biased sex ratio (106 females:40 males), regions of seven cDNA loci were analyzed with single-strand conformation polymorphism (SSCP) analysis to identify and orient linkage groups. Genotypes were also scored at 73 random amplified polymorphic DNA (RAPD)-SSCP loci. In addition to the sex locus, at least four QTL affecting sex determination were detected with interval mapping on linkage groups I and II. Alleles at the sex locus cumulatively accounted for approximately 61 77% of the genetic variance in sex. Alleles at QTL adjacent to the sex locus and at a QTL on the opposite end of linkage group I increased the proportion of females, but alleles at a QTL on linkage group I and a second QTL on linkage group II increased the proportion of males. The female-biased sex ratio observed in laboratory colonies of O. triseriatus is most easily explained by the existence of multiple female biased distorter loci, as have been observed in other Diptera. PMID- 14757729 TI - Genetic structure of Asian populations of Bombus ignitus (Hymenoptera: Apidae). AB - The genetic structure of seven mainland and island Asian populations of Bombus ignitus was investigated using nine microsatellite markers and the sequences of part of the mitochondrial cytochrome b (cytb) gene. While microsatellite markers showed high genetic variability, no sequence variation was found in the cytb gene fragment analyzed. The number of microsatellite alleles ranged from 9 to 24. Gene diversities per locus per population ranged from 0.378 to 0.992. Analysis of molecular variance (AMOVA) and most pairwise F(ST) values showed significant genetic differentiation between mainland and island populations. Cytb sequences data and microsatellite bottleneck tests indicated that almost all populations were subjected to recent bottlenecks. Our results suggest that B. ignitus populations diverged due to recent bottlenecks and geographic isolation. PMID- 14757730 TI - AFLP-based genetic linkage map for the red flour beetle (Tribolium castaneum). AB - The red flour beetle (Tribolium castaneum) is a major pest of stored grain and grain products and a popular model species for a variety of ecological, evolutionary, and developmental biology studies. Development of a linkage map based on reproducible and highly polymorphic molecular markers would greatly facilitate research in these disciplines. We have developed a genetic linkage map using 269 amplified fragment length polymorphism (AFLP) markers. Ten previously known random amplified polymorphic DNA (RAPD) markers were used as anchor markers for linkage group assignment. The linkage map was constructed through genotyping two independent F(2) segregating populations with 48 AFLP primer combinations. Each primer combination generated an average of 4.6 AFLP markers eligible for linkage mapping. The length of the integrated map is 573 cM, giving an average marker resolution of 2.0 cM and an average physical distance per genetic distance of 350 kb/cM. A cluster of loci on linkage group 3 exhibited significant segregation distortion. We have also identified six X-linked and two Y-linked markers. Five mapped AFLP fragments were sequenced and converted to sequence tagged site (STS) markers. PMID- 14757731 TI - A minisatellite with fold-back structure is included in the 5'-flanking region of the Adh gene of Scaptodrosophila lebanonensis. AB - A tandem repetitive sequence with a repeat unit of 12 bp has been found 1.3 kb upstream of the Adh gene of Scaptodrosophila lebanonensis. This repetitive sequence extends over 4.3 kb and consists of two inverted arrays (a fold-back segment). The repeated unit with a consensus sequence GAATACAGAATA is highly conserved and the nucleotide substitutions are not distributed randomly among the 12 bp. In situ hybridization in S. lebanonensis polytene chromosomes revealed two signals, one at the 60A section, the Adh locus, and a second site in the same chromosome at the 60C section close to the telomere. This same pattern of hybridization is obtained in all the analyzed strains including the subspecies S. lebanonensis casteeli. The minisatellite sequence accounts for about 0.03-0.04% of the S. lebanonensis genome and showed intraspecific variability in tandem repeat numbers. Possible functions of this sequence are discussed. PMID- 14757732 TI - Modes of selection and recombination response in Drosophila melanogaster. AB - A selection experiment for sternopleural bristle number in Drosophila melanogaster was undertaken to analyze the correlated effects on recombination. Replicate lines were subjected to divergent directional selection and to stabilizing selection. Recombination rates for markers on chromosomes 2 (dp-cn bw) and 3 (se-ss-ro) were compared to those from a control. All lines responded as predicted for bristle number. Lines selected for both increased and decreased bristle number exhibited significantly increased recombination rates. The predicted recombination response from stabilizing selection is suggested by our data, but only one comparison is statistically significant. These results, taken with other studies, support the proposal that genetic recombination enhances individual fitness when populations are experiencing environmental change. Less conclusively, our results suggest that populations undergoing stabilizing selection may respond by reducing their rates of crossing over. PMID- 14757734 TI - Estimation of allele numbers at the sex-determining locus in a field population of the turnip sawfly (Athalia rosae). AB - Hymenopteran insects (sawflies, ants, bees, and wasps) have an unusual genetic system called haplodiploidy, where parthenogenetically produced haploid eggs become males, and fertilized, diploid eggs become females. Several hypotheses have been proposed to explain the mechanism of such sex determination, including control at a single polymorphic locus. From experiments of mother-son mating and using a genetic marker, we show that a single multiallele locus controls sex determination in the turnip sawfly (Athalia rosae). We estimated the number of alleles at this single locus in a field population by analyzing the rate of diploid males in the field and the rate of diploid males by random crossing in the laboratory. Only one diploid male was discovered in 1306 diploid larvae collected in the field. However, the number of alleles calculated by random crossing in the laboratory was 45-50. We suggest that the effective population size may be much larger than that from the areas where we collected larvae, and that there are mechanisms for avoiding inbreeding, including protogyny, dispersion, and sperm displacement by second-mated males. PMID- 14757733 TI - Inheritance of white head spotting in natural populations of South American water rat (Nectomys squamipes Rodentia: Sigmodontinae). AB - Specimens with white head spots are present at low frequency in the natural populations of South American water rat (Nectomys squamipes) and absent in the sibling species Nectomys rattus. We analyzed the pattern of inheritance of the phenotype using complex segregation analysis of pedigrees of a captive-bred population of N. squamipes. We found that the inheritance of the white head spot in this species can be described within the framework of the major gene recessive model with incomplete penetrance of genotypes. PMID- 14757735 TI - A prototype object database for mitochondrial DNA variation. AB - Surveys of biochemical and molecular genetic variation in natural populations have generated a wealth of data, but this valuable resource has not been adequately preserved. We hope to prevent further loss by establishing a community database for population genetic surveys. We explored the feasibility of a population genetics database by developing a prototype for animal mitochondrial DNA (mtDNA) surveys. This prototype includes the specification of a format for data files that are to be submitted to the database, an open-source object database that encapsulates data with methods to display and analyze data, and a website where data can be retrieved in either its original form or extensible markup language (XML). Data from more than 50 published surveys of mtDNA variation were retrieved from the literature and entered into the database. We hope that the population genetics community will support this project by contributing both data and expertise. PMID- 14757739 TI - Howard Levene remembered. PMID- 14757741 TI - Editing anti-DNA B cells by Vlambdax. AB - Receptor editing is performed by replacement of Vkappa genes that contribute to autoreactivity. In addition, the Ckappa locus can be deleted by Vkappa rearrangement to intronic or 3' of Ckappa RS sequences (also referred to as kappa deletion elements). B cells that delete the Ckappa can then express lambda light chains. However, the lambda locus, either of man or mouse, does not allow V gene replacement. Nor does it appear to be deleted. Therefore, editing of autoreactive lambda B cells may require alternative pathways. We have found that in anti-DNA heavy chain transgenic mice (tgs) VH3H9/56R, B cells that express anti-DNA receptors comprised of lambda1 in association with an anti-DNA heavy chain often coexpress a kappa chain that prevents DNA binding. We speculate that such isotypically included cells may have low anti-DNA receptor densities, a feature that may lead to self-tolerance. Here we describe a mechanism of preventing DNA binding by expression of a rarely used member of the Vlambda family, Vlambdax. The lambdax B cells of the tgs also express CD25 and may represent B cells that have exhausted light chain editing possibilities. PMID- 14757740 TI - Developmental stage, phenotype, and migration distinguish naive- and effector/memory-like CD4+ regulatory T cells. AB - Regulatory T cells (Tregs) fulfill a central role in immune regulation. We reported previously that the integrin alphaEbeta7 discriminates distinct subsets of murine CD4+ regulatory T cells. Use of this marker has now helped to unravel a fundamental dichotomy among regulatory T cells. alphaE-CD25+ cells expressed L selectin and CCR7, enabling recirculation through lymphoid tissues. In contrast, alphaE -positive subsets (CD25+ and CD25-) displayed an effector/memory phenotype expressing high levels of E/P-selectin-binding ligands, multiple adhesion molecules as well as receptors for inflammatory chemokines, allowing efficient migration into inflamed sites. Accordingly, alphaE -expressing cells were found to be the most potent suppressors of inflammatory processes in disease models such as antigen-induced arthritis. PMID- 14757743 TI - Unc119, a novel activator of Lck/Fyn, is essential for T cell activation. AB - The first step in T cell receptor for antigen (TCR) signaling is the activation of the receptor-bound Src kinases, Lck and Fyn. The exact mechanism of this process is unknown. Here, we report that the novel Src homology (SH) 3/SH2 ligand Uncoordinated 119 (Unc119) associates with CD3 and CD4, and activates Lck and Fyn. Unc119 overexpression increases Lck/Fyn activity in T cells. In Unc119 deficient T cells, Lck/Fyn activity is dramatically reduced with concomitant decrease in interleukin 2 production and cellular proliferation. Reconstitution of cells with Unc119 reverses the signaling and functional outcome. Thus, Unc119 is a receptor-associated activator of Src-type kinases. It provides a novel mechanism of signal generation in the TCR complex. PMID- 14757742 TI - WIP regulates signaling via the high affinity receptor for immunoglobulin E in mast cells. AB - Wiskott-Aldrich syndrome protein-interacting protein (WIP) stabilizes actin filaments and is important for immunoreceptor-mediated signal transduction leading to actin cytoskeleton rearrangement in T and B cells. Here we report a role for WIP in signaling pathways downstream of the high affinity receptor for immunoglobulin (Ig)E (FcepsilonRI) in mast cells. WIP-deficient bone marrow derived mast cells (BMMCs) were impaired in their capacity to degranulate and secrete interleukin 6 after FcepsilonRI ligation. Calcium mobilization, phosphorylation of Syk, phospholipase C-g2, and c-Jun NH2-terminal kinase were markedly decreased in WIP-deficient BMMCs. WIP was found to associate with Syk after FcepsilonRI ligation and to inhibit Syk degradation as evidenced by markedly diminished Syk levels in WIP-deficient BMMCs. WIP-deficient BMMCs exhibited no apparent defect in their subcortical actin network and were normal in their ability to form protrusions when exposed to an IgE-coated surface. However, the kinetics of actin changes and the cell shape changes that follow FcepsilonRI signaling were altered in WIP-deficient BMMCs. These results suggest that WIP regulates FcepsilonRI-mediated mast cell activation by regulating Syk levels and actin cytoskeleton rearrangement. PMID- 14757744 TI - Pathogenic profiles and molecular signatures of antinuclear autoantibodies rescued from NZM2410 lupus mice. AB - Two outstanding questions concerning antinuclear antibodies (ANAs) in lupus involve their pathogenic potential and their molecular signatures. To address these questions, a panel of 56 antinuclear and 47 nonnuclear binding monoclonal antibodies was rescued from four seropositive NZM2410 lupus mice. The monoclonals varied in their reactivity to nucleosomes, ssDNA, dsDNA, and glomerular substrate. A large fraction of the antibodies demonstrated apparent polyreactivity (to DNA, histones, and glomerular antigens) due to bound, DNase-1 sensitive nuclear antigenic bridges. Although nephrophilic immunoglobulin (Ig) M and IgG antibodies were the most pathogenic, the dsDNA-binding antibodies were modestly so; in contrast, antinucleosome antibodies were clearly not pathogenic. Compared with the nonnuclear antigen-binding monoclonal antibodies rescued from the same mice, ANAs exhibited increased utilization of VH5/7183 genes and highly cationic heavy chain (HC) CDR3 regions. Most intriguingly, the CDR3 regions of the ANAs exhibited alternating arginine/lysine peaks at H96, H98, and H100, with neutral troughs at H95, H97, and H99. To summarize, glomerular-binding anti-dsDNA antibodies appear to be the most pathogenic variety of lupus autoantibodies. The presence of an alternating charge pattern in their HC CDR3 regions appears to be a prominent hallmark of ANAs. PMID- 14757746 TI - GATA-3 in human T cell helper type 2 development. AB - The delineation of the in vivo role of GATA-3 in human T cell differentiation is a critical step in the understanding of molecular mechanisms directing human immune responses. We examined T cell differentiation and T cell-mediated effector functions in individuals lacking one functional GATA-3 allele. CD4 T cells from GATA-3+/- individuals expressed significantly reduced levels of GATA-3, associated with markedly decreased T helper cell (Th)2 frequencies in vivo and in vitro. Moreover, Th2 cell-mediated effector functions, as assessed by serum levels of Th2-dependent immunoglobulins (Igs; IgG4, IgE), were dramatically decreased, whereas the Th1-dependent IgG1 was elevated compared with GATA-3+/+ controls. Concordant with these data, silencing of GATA-3 in GATA-3+/+ CD4 T cells with small interfering RNA significantly reduced Th2 cell differentiation. Moreover, GATA-3 mRNA levels increased under Th2-inducing conditions and decreased under Th1-inducing conditions. Taken together, the data strongly suggest that GATA-3 is an important transcription factor in regulating human Th2 cell differentiation in vivo. PMID- 14757745 TI - Survivin loss in thymocytes triggers p53-mediated growth arrest and p53 independent cell death. AB - Because survivin-null embryos die at an early embryonic stage, the role of survivin in thymocyte development is unknown. We have investigated the role by deleting the survivin gene only in the T lineage and show here that loss of survivin blocks the transition from CD4- CD8- double negative (DN) thymocytes to CD4+ CD8+ double positive cells. Although the pre-T cell receptor signaling pathway is intact in survivin-deficient thymocytes, the cells cannot respond to its signals. In response to proliferative stimuli, cycling survivin-deficient DN cells exhibit cell cycle arrest, a spindle formation defect, and increased cell death. Strikingly, loss of survivin activates the tumor suppressor p53. However, the developmental defects caused by survivin deficiency cannot be rescued by p53 inactivation or introduction of Bcl-2. These lines of evidence indicate that developing thymocytes depend on the cytoprotective function of survivin and that this function is tightly coupled to cell proliferation but independent of p53 and Bcl-2. Thus, survivin plays a critical role in early thymocyte development. PMID- 14757747 TI - Differential regulation of TCR-mediated gene transcription by Vav family members. AB - Although all three Vav family members are expressed in T lymphocytes, the role that Vav3 plays in T cell activation is poorly defined. Here we show that, like Vav1, Vav3 undergoes rapid tyrosine phosphorylation after T cell receptor (TCR) cross-linkage and interacts with the adaptor molecules SLP76 and 3BP2 in a SH2 dependent manner. However, depletion of Vav1 but not Vav3 protein by RNA interference affects TCR-mediated IL-2 promoter activity. In contrast, Vav3 function is specifically required for coupling TCR stimulation to serum response element-mediated gene transcription. These data indicate that, although both Vav proteins are biochemically coupled to the TCR, they regulate distinct molecular pathways leading to defined gene transcriptional events. PMID- 14757748 TI - Man the nanoscopes. AB - New light microscopy techniques are pushing the limits of resolution to 50 nm and below. Fluorescence microscopy that rivals electron microscopy in resolution but operates on intact cells may be within reach. PMID- 14757749 TI - Oxidative protein folding in eukaryotes: mechanisms and consequences. AB - The endoplasmic reticulum (ER) provides an environment that is highly optimized for oxidative protein folding. Rather than relying on small molecule oxidants like glutathione, it is now clear that disulfide formation is driven by a protein relay involving Ero1, a novel conserved FAD-dependent enzyme, and protein disulfide isomerase (PDI); Ero1 is oxidized by molecular oxygen and in turn acts as a specific oxidant of PDI, which then directly oxidizes disulfide bonds in folding proteins. While providing a robust driving force for disulfide formation, the use of molecular oxygen as the terminal electron acceptor can lead to oxidative stress through the production of reactive oxygen species and oxidized glutathione. How Ero1p distinguishes between the many different PDI-related proteins and how the cell minimizes the effects of oxidative damage from Ero1 remain important open questions. PMID- 14757750 TI - Cleavage furrow positioning. AB - To complete the cell cycle, the cleavage furrow draws the plasma membrane toward the cell center, pinching the cytoplasm into two lobes that are subsequently separated into two cells. The position of the cleavage furrow is induced by the mitotic spindle during early anaphase. Although the mechanism of cleavage furrow positioning is not understood at a molecular level, recent results suggest that it might be mediated by local relief from the inhibitory effects of microtubules. PMID- 14757751 TI - Krox-20 inhibits Jun-NH2-terminal kinase/c-Jun to control Schwann cell proliferation and death. AB - The transcription factor Krox-20 controls Schwann cell myelination. Schwann cells in Krox-20 null mice fail to myelinate, and unlike myelinating Schwann cells, continue to proliferate and are susceptible to death. We find that enforced Krox 20 expression in Schwann cells cell-autonomously inactivates the proliferative response of Schwann cells to the major axonal mitogen beta-neuregulin-1 and the death response to TGFbeta or serum deprivation. Even in 3T3 fibroblasts, Krox-20 not only blocks proliferation and death but also activates the myelin genes periaxin and protein zero, showing properties in common with master regulatory genes in other cell types. Significantly, a major function of Krox-20 is to suppress the c-Jun NH2-terminal protein kinase (JNK)-c-Jun pathway, activation of which is required for both proliferation and death. Thus, Krox-20 can coordinately control suppression of mitogenic and death responses. Krox-20 also up-regulates the scaffold protein JNK-interacting protein 1 (JIP-1). We propose this as a possible component of the mechanism by which Krox-20 regulates JNK activity during Schwann cell development. PMID- 14757752 TI - A novel LIM protein Cal promotes cardiac differentiation by association with CSX/NKX2-5. AB - The cardiac homeobox transcription factor CSX/NKX2-5 plays an important role in vertebrate heart development. Using a yeast two-hybrid screening, we identified a novel LIM domain-containing protein, named CSX-associated LIM protein (Cal), that interacts with CSX/NKX2-5. CSX/NKX2-5 and Cal associate with each other both in vivo and in vitro, and the LIM domains of Cal and the homeodomain of CSX/NKX2-5 were necessary for mutual binding. Cal itself possessed the transcription promoting activity, and cotransfection of Cal enhanced CSX/NKX2-5-induced activation of atrial natriuretic peptide gene promoter. Cal contained a functional nuclear export signal and shuttled from the cytoplasm into the nucleus in response to calcium. Accumulation of Cal in the nucleus of P19CL6 cells promoted myocardial cell differentiation accompanied by increased expression levels of the target genes of CSX/NKX2-5. These results suggest that a novel LIM protein Cal induces cardiomyocyte differentiation through its dynamic intracellular shuttling and association with CSX/NKX2-5. PMID- 14757753 TI - Clustering of Nck by a 12-residue Tir phosphopeptide is sufficient to trigger localized actin assembly. AB - Enteropathogenic Escherichia coli (EPEC) translocates effector proteins into mammalian cells to promote reorganization of the cytoskeleton into filamentous actin pedestals. One effector, Tir, is a transmembrane receptor for the bacterial surface adhesin intimin, and intimin binding by the extracellular domain of Tir is required for actin assembly. The cytoplasmic NH2 terminus of Tir interacts with focal adhesion proteins, and its tyrosine-phosphorylated COOH terminus binds Nck, a host adaptor protein critical for pedestal formation. To define the minimal requirements for EPEC-mediated actin assembly, Tir derivatives were expressed in mammalian cells in the absence of all other EPEC components. Replacement of the NH2 terminus of Tir with a viral membrane-targeting sequence promoted efficient surface expression of a COOH-terminal Tir fragment. Artificial clustering of this fusion protein revealed that the COOH terminus of Tir, by itself, is sufficient to initiate a complete signaling cascade leading to pedestal formation. Consistent with this finding, clustering of Nck by a 12 residue Tir phosphopeptide triggered actin tail formation in Xenopus egg extracts. PMID- 14757754 TI - Distinct roles of MLCK and ROCK in the regulation of membrane protrusions and focal adhesion dynamics during cell migration of fibroblasts. AB - We examined the role of regulatory myosin light chain (MLC) phosphorylation of myosin II in cell migration of fibroblasts. Myosin light chain kinase (MLCK) inhibition blocked MLC phosphorylation at the cell periphery, but not in the center. MLCK-inhibited cells did not assemble zyxin-containing adhesions at the periphery, but maintained focal adhesions in the center. They generated membrane protrusions all around the cell, turned more frequently, and migrated less effectively. In contrast, Rho-associated kinase (ROCK) inhibition blocked MLC phosphorylation in the center, but not at the periphery. ROCK-inhibited cells assembled zyxin-containing adhesions at the periphery, but not focal adhesions in the center. They moved faster and more straight. On the other hand, inhibition of myosin phosphatase increased MLC phosphorylation and blocked peripheral membrane ruffling, as well as turnover of focal adhesions and cell migration. Our results suggest that myosin II activated by MLCK at the cell periphery controls membrane ruffling, and that the spatial regulation of MLC phosphorylation plays critical roles in controlling cell migration of fibroblasts. PMID- 14757756 TI - Role of protein kinase C isoforms in the regulation of interleukin-13-induced 15 lipoxygenase gene expression in human monocytes. AB - We reported previously that interleukin-13 (IL-13) induces tyrosine phosphorylation/activation of Jak2 and Tyk2 kinases and Stats 1, 3, 5, and 6 in primary human monocytes. We recently revealed that p38 MAPK-mediated serine phosphorylation of both Stat1 and Stat3 is required for the induction of 15 lipoxygenase (15-LO) expression by IL-13. In this study, we present data indicating that another serine/threonine kinase, PKCdelta, is also required for IL-13-induced 15-LO expression. PKCdelta, a member of the novel protein kinase C (PKC) subclass, was rapidly phosphorylated and activated upon exposure to IL-13. Treatment of cells with rottlerin, a PKCdelta inhibitor, blocked IL-13-induced 15 LO mRNA and protein expression, whereas Go6976, an inhibitor of the conventional PKC subclass, had no inhibitory effects. Down-regulation of cellular PKCdelta protein levels by PKCdelta-specific antisense oligodeoxyribonucleotides also inhibited 15-LO expression markedly. IL-13-induced 15-LO expression resulted in significant inhibition of synthesis of the potent chemotactic factor leukotriene B4, and that process was reversed by rottlerin, presumably through the blockage of PKCdelta-dependent 15-LO expression. Furthermore, our data demonstrate that IL 13-mediated activation of PKCdelta and p38 MAPK are independent pathways, because inhibition of one kinase activity had no effect on the other, suggesting that the two pathways act in parallel to regulate the downstream targets necessary for 15 LO expression. Inhibition of PKCdelta activation by rottlerin also markedly attenuated IL-13-induced Stat3 DNA binding activity. Our findings indicate that PKCdelta plays an important role in regulating IL-13-induced 15-LO expression in human monocytes and subsequently modulates the inflammatory responses mediated by 15-LO products. PMID- 14757755 TI - Rap1 up-regulation and activation on plasma membrane regulates T cell adhesion. AB - Rap1 and Ras are closely related GTPases that share some effectors but have distinct functions. We studied the subcellular localization of Rap1 and its sites of activation in living cells. Both GFP-tagged Rap1 and endogenous Rap1 were localized to the plasma membrane (PM) and endosomes. The PM association of GFP Rap1 was dependent on GTP binding, and GFP-Rap1 was rapidly up-regulated on this compartment in response to mitogens, a process blocked by inhibitors of endosome recycling. A novel fluorescent probe for GTP-bound Rap1 revealed that this GTPase was transiently activated only on the PM of both fibroblasts and T cells. Activation on the PM was blocked by inhibitors of endosome recycling. Moreover, inhibition of endosome recycling blocked the ability of Rap1 to promote integrin mediated adhesion of T cells. Thus, unlike Ras, the membrane localizations of Rap1 are dynamically regulated, and the PM is the principle platform from which Rap1 signaling emanates. These observations may explain some of the biological differences between these GTPases. PMID- 14757757 TI - Role of the N-terminal catalytic domain of angiotensin-converting enzyme investigated by targeted inactivation in mice. AB - Angiotensin-converting enzyme (ACE) produces the vasoconstrictor angiotensin II. The ACE protein is composed of two homologous domains, each binding zinc and each independently catalytic. To assess the physiologic significance of the two ACE catalytic domains, we used gene targeting in mice to introduce two point mutations (H395K and H399K) that selectively inactivated the ACE N-terminal catalytic site. This modification does not affect C-terminal enzymatic activity or ACE protein expression. In addition, the testis ACE isozyme is not affected by the mutations. Analysis of homozygous mutant mice (termed ACE 7/7) showed normal plasma levels of angiotensin II but an elevation of plasma and urine N-acetyl-Ser Asp-Lys-Pro, a peptide suggested to inhibit bone marrow maturation. Despite this, ACE 7/7 mice had blood pressure, renal function, and hematocrit that were indistinguishable from wild-type mice. We also studied compound heterozygous mice in which one ACE allele was null (no ACE expression) and the second allele encoded the mutations selectively inactivating the N-terminal catalytic domain. These mice produced approximately half the normal levels of ACE, with the ACE protein lacking N-terminal catalytic activity. Despite this, the mice have a phenotype indistinguishable from wild-type animals. This study shows that, in vivo, the presence of the C-terminal ACE catalytic domain is sufficient to maintain a functional renin-angiotensin system. It also strongly suggests that the anemia present in ACE null mice is not due to the accumulation of the peptide N-acetyl-Ser-Asp-Lys-Pro. PMID- 14757758 TI - Monoclonal antibodies specific for the empty conformation of HLA-DR1 reveal aspects of the conformational change associated with peptide binding. AB - Class II major histocompatibility complex (MHC) proteins bind peptides and present them at the cell surface for interaction with CD4+ T cells as part of the system by which the immune system surveys the body for signs of infection. Peptide binding is known to induce conformational changes in class II MHC proteins on the basis of a variety of hydrodynamic and spectroscopic approaches, but the changes have not been clearly localized within the overall class II MHC structure. To map the peptide-induced conformational change for HLA-DR1, a common human class II MHC variant, we generated a series of monoclonal antibodies recognizing the beta subunit that are specific for the empty conformation. Each antibody reacted with the empty but not the peptide-loaded form, for both soluble recombinant protein and native protein expressed at the cell surface. Antibody binding epitopes were characterized using overlapping peptides and alanine scanning substitutions and were localized to two distinct regions of the protein. The pattern of key residues within the epitopes suggested that the two epitope regions undergo substantial conformational alteration during peptide binding. These results illuminate aspects of the structure of the empty forms and the nature of the peptide-induced conformational change. PMID- 14757759 TI - Lateral membrane biogenesis in human bronchial epithelial cells requires 190-kDa ankyrin-G. AB - Ankyrin-G polypeptides are required for restriction of voltage-gated sodium channels, L1 cell adhesion molecules, and beta IV spectrin to axon initial segments and are believed to couple the Na/K-ATPase to the spectrin-actin network at the lateral membrane in epithelial cells. We report here that depletion of 190 kDa ankyrin-G in human bronchial epithelial cells by small interfering RNA results in nearly complete loss of lateral plasma membrane in interphase cells, and also blocks de novo lateral membrane biogenesis following mitosis. Loss of the lateral membrane domain is accompanied by an expansion of apical and basal plasma membranes and preservation of apical-basal polarity. Expression of rat 190 kDa ankyrin-G, which is resistant to human small interfering RNA, prevents loss of the lateral membrane following depletion of human 190-kDa ankyrin-G. Human 220 kDa ankyrin-B, a closely related ankyrin isoform, is incapable of preserving the lateral membrane following 190-kDa ankyrin-G depletion. Moreover, analysis of rat 190-kDa ankyrin G/ankyrin B chimeras shows that all three domains of 190-kDa ankyrin-G are required for preservation of the lateral membrane. These results demonstrate that 190-kDa ankyrin-G plays a pleiotropic role in assembly of lateral membranes of bronchial epithelial cells. PMID- 14757760 TI - Thr-90 plays a vital role in the structure and function of bacteriorhodopsin. AB - The role of Thr-90 in the bacteriorhodopsin structure and function was investigated by its replacement with Ala and Val. The mutant D115A was also studied because Asp-115 in helix D forms a hydrogen bond with Thr-90 in helix C. Differential scanning calorimetry showed a decreased thermal stability of all three mutants, with T90A being the least stable. Light-dark adaptation of T90A was found to be abnormal and salt-dependent. Proton transport monitored using pyranine signals was approximately 10% of wild type for T90A, 20% for T90V, and 50% for D115A. At neutral or alkaline pH, the M rise of these mutants was faster than that of wild type, whereas M decay was slower in T90A. Overall, Fourier transform infrared (FTIR) difference spectra of T90A were strongly pH-dependent. Spectra recorded on films adjusted at the same pH at 243 or 277 K, dry or wet, showed similar features. The D115A and T90V FTIR spectra were closer to WT, showing minor structural differences. The band at 1734 cm(-1) of the deconvoluted FTIR spectrum, corresponding to the carboxylate of Asp-115, was absent in all mutants. In conclusion, Thr-90 plays a critical role in maintaining the operative location and structure of helix C through three complementary interactions, namely an interhelical hydrogen bond with Asp-115, an intrahelical hydrogen bond with the peptide carbonyl oxygen of Trp-86, and a steric contact with the retinal. The interactions established by Thr-90 emerge as a general feature of archaeal rhodopsin proteins. PMID- 14757761 TI - Kinetic properties and ammonium-dependent regulation of cytosolic isoenzymes of glutamine synthetase in Arabidopsis. AB - Glutamine synthetase (GS; EC 6.3.1.2) is a key enzyme of nitrogen assimilation, catalyzing the synthesis of glutamine from ammonium and glutamate. In Arabidopsis, cytosolic GS (GS1) was accumulated in roots when plants were excessively supplied with ammonium; however, the GS activity was controlled at a constant level. The discrepancy between the protein content and enzyme activity of GS1 was attributable to the kinetic properties and expression of four distinct isoenzymes encoded by GLN1;1, GLN1;2, GLN1;3 and GLN1;4, genes that function complementary to each other in Arabidopsis roots. GLN1;2 was the only isoenzyme significantly up-regulated by ammonium, which correlated with the rapid increase in total GS1 protein. GLN1;2 was localized in the vasculature and exhibited low affinities to ammonium (Km = 2450 +/- 150 microm) and glutamate (Km = 3.8 +/- 0.2 mm). The expression of the counterpart vascular tissue-localizing low affinity isoenzyme, GLN1;3, was not stimulated by ammonium; however, the enzyme activity of GLN1;3 was significantly inhibited by a high concentration of glutamate. By contrast, the high affinity isoenzyme, GLN1;1 (Km for ammonium < 10 microm; Km for glutamate = 1.1 +/- 0.4 mm) was abundantly accumulated in the surface layers of roots during nitrogen limitation and was down-regulated by ammonium excess. GLN1;4 was another high affinity-type GS1 expressed in nitrogen-starved plants but was 10-fold less abundant than GLN1;1. These results suggested that dynamic regulations of high and low affinity GS1 isoenzymes at the levels of mRNA and enzyme activities are dependent on nitrogen availabilities and may contribute to the homeostatic control of glutamine synthesis in Arabidopsis roots. PMID- 14757762 TI - An evolutionary and molecular analysis of Bmp2 expression. AB - The coding regions of many metazoan genes are highly similar. For example, homologs to the key developmental factor bone morphogenetic protein (BMP) 2 have been cloned by sequence identity from arthropods, mollusks, cnidarians, and nematodes. Wide conservation of protein sequences suggests that differential gene expression explains many of the vast morphological differences between species. To test the hypothesis that the regulatory mechanisms controlling this evolutionarily ancient and critical gene are conserved, we compared sequences flanking Bmp2 genes of several species. We identified numerous conserved noncoding sequences including some retained because the fish lineage separated 450 million years ago. We tested the function of some of these sequences in the F9 cell model system of Bmp2 expression. We demonstrated that both mouse and primate Bmp2 promoters drive a reporter gene in an expression pattern resembling that of the endogenous transcript in F9 cells. A conserved Sp1 site contributes to the retinoic acid responsiveness of the Bmp2 promoter, which lacks a classical retinoic acid response element. We have also discovered a sequence downstream of the stop codon whose conservation between humans, rodents, deer, chickens, frogs, and fish is striking. A fragment containing this region influences reporter gene expression in F9 cells. The conserved region contains elements that may mediate the half-life of the Bmp2 transcript. Together, our molecular and evolutionary analysis has identified new regulatory elements controlling Bmp2 expression. PMID- 14757763 TI - Upstream stimulatory factor (USF) proteins induce human TGF-beta1 gene activation via the glucose-response element-1013/-1002 in mesangial cells: up-regulation of USF activity by the hexosamine biosynthetic pathway. AB - The hyperglycemia-enhanced flux through the hexosamine biosynthetic pathway (HBP) has been implicated in the up-regulated gene expression of transforming growth factor-beta1 (TGF-beta1) in mesangial cells, thus leading to mesangial matrix expansion and diabetic glomerulosclerosis. Since the -1013 to -1002 region of the TGF-beta1 promoter shows high homology to glucose-response elements (GlRE) formerly described in genes involved in glucose metabolism, we studied the function of the GlRE in the high glucose-induced TGF-beta1 gene activation in mesangial cells. We found that high glucose concentrations enhanced the nuclear amount of upstream stimulatory factors (USF) and their binding to this sequence. Fusion of the GlRE to the thymidine kinase promoter resulted in glucose responsiveness of this promoter construct. Overexpression of either USF-1 or USF 2 increased TGF-beta1 promoter activity 2-fold, which was prevented by mutation or deletion of the GlRE. The high glucose-induced activation of the GlRE is mediated by the HBP; increased flux through the HBP induced by high glucose concentrations, by glutamine, or by overexpression of the rate-limiting enzyme glutamine:fructose-6-phosphate aminotransferase (GFAT) particularly activated USF 2 expression. GFAT-overexpressing cells showed higher USF binding activity to the GlRE and enhanced promoter activation via the GlRE. Increasing O-GlcNAc modification of proteins by streptozotocin, thereby mimicking HBP activation, also resulted in increased mRNA and nuclear protein levels of USF-2, leading to enhanced DNA binding activity to the GlRE. USF proteins themselves were not found to be O-GlcNAc-modified. Thus, we have provided evidence for a new molecular mechanism linking high glucose-enhanced HBP activity with increased nuclear USF protein levels and DNA binding activity and with up-regulated TGF-beta1 promoter activity. PMID- 14757764 TI - Glucosylceramidase mass and subcellular localization are modulated by cholesterol in Niemann-Pick disease type C. AB - Niemann-Pick disease type C (NPC) is characterized by the accumulation of cholesterol and sphingolipids in the late endosomal/lysosomal compartment. The mechanism by which the concentration of sphingolipids such as glucosylceramide is increased in this disease is poorly understood. We have found that, in NPC fibroblasts, the cholesterol storage affects the stability of glucosylceramidase (GCase), decreasing its mass and activity; a reduction of cholesterol raises the level of GCase to nearly normal values. GCase is activated and stabilized by saposin C (Sap C) and anionic phospholipids. Here we show by immunofluorescence microscopy that in normal fibroblasts, GCase, Sap C, and lysobisphosphatidic acid (LBPA), the most abundant anionic phospholipid in the endolysosomal system, reside in the same intracellular vesicular structures. In contrast, the colocalization of GCase, Sap C, and LBPA is markedly impaired in NPC fibroblasts but can be re-established by cholesterol depletion. These data show for the first time that the level of cholesterol modulates the interaction of GCase with its protein and lipid activators, namely Sap C and LBPA, regulating the GCase activity and stability. PMID- 14757765 TI - An intestinal parasitic protist, Entamoeba histolytica, possesses a non-redundant nitrogen fixation-like system for iron-sulfur cluster assembly under anaerobic conditions. AB - We have characterized the iron-sulfur (Fe-S) cluster formation in an anaerobic amitochondrial protozoan parasite, Entamoeba histolytica, in which Fe-S proteins play an important role in energy metabolism and electron transfer. A genomewide search showed that E. histolytica apparently possesses a simplified and non redundant NIF (nitrogen fixation)-like system for the Fe-S cluster formation, composed of only a catalytic component, NifS, and a scaffold component, NifU. Amino acid alignment and phylogenetic analyses revealed that both amebic NifS and NifU (EhNifS and EhNifU, respectively) showed a close kinship to orthologs from epsilon-proteobacteria, suggesting that both of these genes were likely transferred by lateral gene transfer from an ancestor of epsilon-proteobacteria to E. histolytica. The EhNifS protein expressed in E. coli was present as a homodimer, showing cysteine desulfurase activity with a very basic optimum pH compared with NifS from other organisms. Eh-NifU protein existed as a tetramer and contained one stable [2Fe-2S]2+ cluster per monomer, revealed by spectroscopic and iron analyses. Fractionation of the whole parasite lysate by anion exchange chromatography revealed three major cysteine desulfurase activities, one of which corresponded to the EhNifS protein, verified by immunoblot analysis using the specific EhNifS antibody; the other two peaks corresponded to methionine gamma-lyase and cysteine synthase. Finally, ectopic expression of the EhNifS and EhNifU genes successfully complemented, under anaerobic but not aerobic conditions, the growth defect of an Escherichia coli strain, in which both the isc and suf operons were deleted, suggesting that EhNifS and EhNifU are necessary and sufficient for Fe-S clusters of non nitrogenase Fe-S proteins to form under anaerobic conditions. This is the first demonstration of the presence and biological significance of the NIF-like system in eukaryotes. PMID- 14757766 TI - Thiamine biosynthesis in Escherichia coli: in vitro reconstitution of the thiazole synthase activity. AB - The biosynthesis of thiamine in Escherichia coli requires the formation of an intermediate thiazole from tyrosine, 1-deoxy-d-xylulose-5-phosphate (Dxp), and cysteine using at least six structural proteins, ThiFSGH, IscS, and ThiI. We describe for the first time the reconstitution of thiazole synthase activity using cell-free extracts and proteins derived from adenosine-treated E. coli 83-1 cells. The addition of adenosine or adenine to growing cultures of Aerobacter aerogenes, Salmonella typhimurium, and E. coli has been shown previously to relieve the repression by thiamine of its own biosynthesis and increase the expression levels of the thiamine biosynthetic enzymes. By exploiting this effect, we show that the in vitro thiazole synthase activity of cleared lysates or desalted proteins from E. coli 83-1 cells is dependent upon the addition of purified ThiGH-His complex, tyrosine (but not cysteine or 1-deoxy-d-xylulose-5 phosphate), and an as yet unidentified intermediate present in the protein fraction from these cells. The activity is strongly stimulated by the addition of S-adenosylmethionine and NADPH. PMID- 14757768 TI - C-3 epimerization of vitamin D3 metabolites and further metabolism of C-3 epimers: 25-hydroxyvitamin D3 is metabolized to 3-epi-25-hydroxyvitamin D3 and subsequently metabolized through C-1alpha or C-24 hydroxylation. AB - Recently, it was revealed that 1alpha,25-dihydroxyvitamin D3 (1alpha,25(OH)2D3) and 24R,25-dihydroxyvitamin D3 (24,25(OH)2D3) were metabolized to their respective epimers of the hydroxyl group at C-3 of the A-ring. We now report the isolation and structural assignment of 3-epi-25-hydroxyvitamin D3 (3-epi-25(OH)D3 as a major metabolite of 25-hydroxyvitamin D3 (25(OH)D3) and the further metabolism of C-3 epimers of vitamin D3 metabolites. When 25(OH)D3 was incubated with various cultured cells including osteosarcoma, colon adenocarcinoma, and hepatoblastoma cell lines, 3-epi-25(OH)D3 and 24,25 (OH)2D3 were commonly observed as a major and minor metabolite of 25(OH)D3, respectively. 25(OH)D3 was at least as sensitive to C-3 epimerization as 1alpha, 25(OH)2D3 which has been reported as a substrate for the C-3 epimerization reaction. Unlike these cultured cells, LLC-PK1 cells, a porcine kidney cell line, preferentially produced 24,25(OH)2D3 rather than 3-epi-25(OH)D3. We also confirmed the existence of 3-epi 25(OH)D3 in the serum of rats intravenously given pharmacological doses of 25(OH)D3. The cultured cells metabolized 3-epi-25OHD3 and 3-epi-1alpha,25(OH)2D3 to 3-epi-24,25(OH)2D3 and 3-epi-1alpha,24,25(OH)3D3, respectively. In addition, we demonstrated that 3-epi-25(OH)D3 was metabolized to 3-epi-1alpha,25(OH)2D3 by CYP27B1 and to 3-epi-24,25(OH)2D3 by CYP24 using recombinant Escherichia coli cell systems. 3-Epi-25(OH)D3, 3-epi-1alpha,25(OH)2D3, and 3-epi-24,25(OH)2D3 were biologically less active than 25(OH)D3, 1alpha,25(OH)2D3, and 24,25(OH)2D3, but 3 epi-1alpha,25(OH)2D3 showed to some extent transcriptional activity toward target genes and anti-proliferative/differentiation-inducing activity against human myeloid leukemia cells (HL-60). These results indicate that C-3 epimerization may be a common metabolic pathway for the major metabolites of vitamin D3. PMID- 14757767 TI - Tazobactam inactivation of SHV-1 and the inhibitor-resistant Ser130 -->Gly SHV-1 beta-lactamase: insights into the mechanism of inhibition. AB - The increasing number of bacteria resistant to combinations of beta-lactam and beta-lactamase inhibitors is creating great difficulties in the treatment of serious hospital-acquired infections. Understanding the mechanisms and structural basis for the inactivation of these inhibitor-resistant beta-lactamases provides a rationale for the design of novel compounds. In the present work, SHV-1 and the Ser(130) --> Gly inhibitor-resistant variant of SHV-1 beta-lactamase were inactivated with tazobactam, a potent class A beta-lactamase inhibitor. Apoenzymes and inhibited beta-lactamases were analyzed by liquid chromatography electrospray ionization mass spectrometry (LC-ESI/MS), digested with trypsin, and the products resolved using LC-ESI/MS and matrix-assisted laser desorption ionization-time of flight mass spectrometry. The mass increases observed for SHV 1 and Ser(130) --> Gly (+ Delta 88 Da and + Delta 70 Da, respectively) suggest that fragmentation of tazobactam readily occurs in the inhibitor-resistant variant to yield an inactive beta-lactamase. These two mass increments are consistent with the formation of an aldehyde (+ Delta 70 Da) and a hydrated aldehyde (+ Delta 88 Da) as stable products of inhibition. Our results reveal that the Ser --> Gly substitution at amino acid position 130 is not essential for enzyme inactivation. By examining the inhibitor-resistant Ser(130) --> Gly beta lactamase, our data are the first to show that tazobactam undergoes fragmentation while still attached to the active site Ser(70) in this enzyme. After acylation of tazobactam by Ser(130) --> Gly, inactivation proceeds independent of any additional covalent interactions. PMID- 14757769 TI - Structural requirements for additional N-linked carbohydrate on recombinant human erythropoietin. AB - N-Linked glycosylation is a post-translational event whereby carbohydrates are added to secreted proteins at the consensus sequence Asn-Xaa-Ser/Thr, where Xaa is any amino acid except proline. Some consensus sequences in secreted proteins are not glycosylated, indicating that consensus sequences are necessary but not sufficient for glycosylation. In order to understand the structural rules for N linked glycosylation, we introduced N-linked consensus sequences by site-directed mutagenesis into the polypeptide chain of the recombinant human erythropoietin molecule. Some regions of the polypeptide chain supported N-linked glycosylation more effectively than others. N-Linked glycosylation was inhibited by an adjacent proline suggesting that sequence context of a consensus sequence could affect glycosylation. One N-linked consensus sequence (Asn123-Thr125) introduced into a position close to the existing O-glycosylation site (Ser126) had an additional O linked carbohydrate chain and not an additional N-linked carbohydrate chain suggesting that structural requirements in this region favored O-glycosylation over N-glycosylation. The presence of a consensus sequence on the protein surface of the folded molecule did not appear to be a prerequisite for oligosaccharide addition. However, it was noted that recombinant human erythropoietin analogs that were hyperglycosylated at sites that were normally buried had altered protein structures. This suggests that carbohydrate addition precedes polypeptide folding. PMID- 14757770 TI - NEDD8 ultimate buster-1L interacts with the ubiquitin-like protein FAT10 and accelerates its degradation. AB - FAT10 is an interferon-gamma-inducible ubiquitin-like protein that consists of two ubiquitin-like domains. FAT10 bears a diglycine motif at its C terminus that can form isopeptide bonds to so far unidentified target proteins. Recently we found that FAT10 and its conjugates are rapidly degraded by the proteasome and that the N-terminal fusion of FAT10 to a long lived protein markedly reduces its half-life. FAT10 may hence direct target proteins to the proteasome for degradation. In this study we report a new interaction partner of FAT10 that may link FAT10 to the proteasome. A yeast two-hybrid screen identified NEDD8 ultimate buster-1L (NUB1L) as a non-covalent binding partner of FAT10, and this interaction was confirmed by coimmunoprecipitation and glutathione S-transferase pull-down experiments. NUB1L is also an interferon-inducible protein that has been reported to interact with the ubiquitin-like protein NEDD8, thus leading to accelerated NEDD8 degradation. Here we show that NUB1L binds to FAT10 much stronger than to NEDD8 and that NEDD8 cannot compete with FAT10 for NUB1L binding. The interaction of FAT10 and NUB1L is specific as green fluorescent fusion proteins containing ubiquitin or SUMO-1 do not bind to NUB1L. The coexpression of NUB1L enhanced the degradation rate of FAT10 8-fold, whereas NEDD8 degradation was only accelerated 2-fold. Because NUB1 was shown to bind to the proteasome subunit RPN10 in vitro and to be contained in 26 S proteasome preparations, it may function as a linker that targets FAT10 for degradation by the proteasome. PMID- 14757772 TI - Identification of the amino acid residues of the platelet glycoprotein Ib (GPIb) essential for the von Willebrand factor binding by clustered charged-to-alanine scanning mutagenesis. AB - At the site of vascular injury, von Willebrand factor (VWF) mediates platelet adhesion to subendothelial connective tissue through binding to the N-terminal domain of the alpha chain of platelet glycoprotein Ib (GPIbalpha). To elucidate the molecular mechanisms of the binding, we have employed charged-to-alanine scanning mutagenesis of the soluble fragment containing the N-terminal 287 amino acids of GPIbalpha. Sixty-two charged amino acids were changed singly or in small clusters, and 38 mutant constructs were expressed in the supernatant of 293T cells. Each mutant was assayed for binding to several monoclonal antibodies for human GPIbalpha and for ristocetin-induced and botrocetin-induced binding of 125I labeled human VWF. Mutations at Glu128, Glu172, and Asp175 specifically decreased both ristocetin- and botrocetin-induced VWF binding, suggesting that these sites are important for VWF binding of platelet GPIb. Monoclonal antibody 6D1 inhibited ristocetin- and botrocetin-induced VWF binding, and a mutation at Glu125 specifically reduced the binding to 6D1. In contrast, antibody HPL7 had no effect for VWF binding, and mutant E121A reduced the HPL7 binding. Mutations at His12 and Glu14 decreased the ristocetin-induced VWF binding with normal botrocetin induced binding. Crystallographic modeling of the VWF-GPIbalpha complex indicated that Glu128 and Asp175 form VWF binding sites; the binding of 6D1 to Glu125 interrupts the VWF binding of Glu128, but HPL7 binding to Glu121 has no effect on VWF binding. Moreover, His12 and Glu14 contact with Glu613 and Arg571 of VWF A1 domain, whose mutations had shown similar phenotype. These findings indicated the novel binding sites required for VWF binding of human GPIbalpha. PMID- 14757771 TI - Potential role of methionine sulfoxide in the inactivation of the chaperone GroEL by hypochlorous acid (HOCl) and peroxynitrite (ONOO-). AB - GroEL is an Escherichia coli molecular chaperone that functions in vivo to fold newly synthesized polypeptides as well as to bind and refold denatured proteins during stress. This protein is a suitable model for its eukaryotic homolog, heat shock protein 60 (Hsp60), due to the high number of conserved amino acid sequences and similar function. Here, we will provide evidence that GroEL is rather insensitive to oxidants produced endogenously during metabolism, such as nitric oxide (.NO) or hydrogen peroxide (H(2)O(2)), but is modified and inactivated by efficiently reactive species generated by phagocytes, such as peroxynitrite (ONOO(-)) and hypochlorous acid (HOCl). For the exposure of 17.5 microm GroEL to 100-250 microm HOCl, the major pathway of inactivation was through the oxidation of methionine to methionine sulfoxide, established through mass spectrometric detection of methionine sulfoxide and the reactivation of a significant fraction of inactivated GroEL by the enzyme methionine sulfoxide reductase B/A (MsrB/A). In addition to the oxidation of methionine, HOCl caused the conversion of cysteine to cysteic acid and this product may account for the remainder of inactivated GroEL not recoverable through MsrB/A. In contrast, HOCl produced only negligible yields of 3-chlorotyrosine. A remarkable finding was the conversion of Met(111) and Met(114) to Met sulfone, which suggests a rather low reduction potential of these 2 residues in GroEL. The high sensitivity of GroEL toward HOCl and ONOO(-) suggests that this protein may be a target for bacterial killing by phagocytes. PMID- 14757773 TI - Activation of RAW264.7 macrophages by bacterial DNA and lipopolysaccharide increases cell surface DNA binding and internalization. AB - Bacterial DNA containing unmethylated CpG motifs is a pathogen-associated molecular pattern (PAMP) that interacts with host immune cells via a toll-like receptor (TLR) to induce immune responses. DNA binding and internalization into cells is independent of TLR expression, receptor-mediated, and required for cell activation. The objective of this study was to determine whether exposure of immune cells to bacterial DNA affects DNA binding and internalization. Treatment of RAW264.7 cells with CpG oligodeoxynucleotide (ODN) for both 18 and 42 h resulted in a significant increase in DNA binding, whereas non-CpG ODN had no effect on DNA binding. Enhanced DNA binding was non-sequence-specific, inhibited by unlabeled DNA, showed saturation, was consistent with increased cell surface DNA receptors, and resulted in enhanced internalization of DNA. Treatment with Escherichia coli DNA or lipopolysaccharide (LPS) also resulted in a significant increase in DNA binding, but treatment with interleukin-1alpha, tumor necrosis factor-alpha, or phorbol 12-myristate 13-acetate had no effect on DNA binding. Soluble factors produced in response to treatment with CpG ODN or LPS did not affect DNA binding. These studies demonstrate that one consequence of activating the host innate immune response by bacterial infection is enhanced binding and internalization of DNA. During this period of increased DNA internalization, RAW264.7 cells were hypo-responsive to continued stimulation by CpG ODN, as assessed by tumor necrosis factor-alpha activity. We speculate the biological significance of increasing DNA binding and internalization following interaction with bacterial PAMPs may provide a mechanism to limit an ongoing immune inflammatory response by enhancing clearance of bacterial DNA from the extracellular environment. PMID- 14757774 TI - Selective increase in renal arcuate innervation density and neurogenic constriction in chronic angiotensin II-infused rats. AB - This study investigated the effects of angiotensin II "slow pressor" hypertension on structure and function of nerves supplying the renal vasculature. Low-dose angiotensin II (10 ng/kg per minute, initially sub-pressor) or saline vehicle was infused intravenously for 21 days in rats, and the effects were compared in renal and mesenteric arteries. Mean arterial pressure averaged 12+/-2 mm Hg higher than in vehicle-infused rats at 21 days. Using electron microscopy, the innervation density of renal arcuate, but not mesenteric arteries of equivalent size, was significantly higher in angiotensin II-infused than in vehicle-infused rats. Functional testing on a pressure myograph revealed that constrictions evoked by nerve stimulation in arcuate arteries were 2.3+/-0.7-fold greater in vessels from angiotensin II-infused compared with vehicle-infused rats (P<0.0001), whereas there was no significant difference in nerve-induced constrictions in mesenteric arteries. Sensitivity to and maximum amplitude of constrictions evoked by phenylephrine were not different in renal or mesenteric arteries between groups, suggesting that the increased neurally evoked constriction in renal arcuate arteries was not caused by postsynaptic changes. Endothelium-dependent vasorelaxation and the vessel wall physical properties were not different between the two groups in either artery. Thus, angiotensin II infusion appeared to evoke renal-specific increases in vessel innervation and increased vasoconstriction to nerve stimulation. These changes appear early and occur before changes in renal endothelial function are apparent. Thus, "slow pressor" angiotensin II hypertension is associated with increased renal innervation, compatible with a pathogenetic role. PMID- 14757775 TI - Pulse pressure is an independent predictor for the progression of aortic wall calcification in patients with controlled hyperlipidemia. AB - Recent epidemiological studies suggested that calcifications of the aorta and the coronary arteries are important predictors for cardiovascular morbidity and mortality. However, the relation between blood pressure components and the progression of vascular wall calcification has remained unclear. We quantified calcium deposits in the abdominal aorta as the percentage of aortic calcification volume (%ACV) using computed tomography in patients with hyperlipidemia. Those who had aortic calcification were treated with lipid-lowering agents and followed up for >2 years (6.3+/-3.2 years). The relationship between the components of blood pressure and the increase in %ACV per year (Delta%ACV/year) was assessed in subjects in whom serum lipid levels were well controlled during the follow-up periods. An age- and sex-adjusted correlation analysis showed that Delta%ACV/year was significantly correlated to body mass index (r=0.229, P=0.015), systolic blood pressure (r=0.244, P=0.009), and pulse pressure (r=0.359, P<0.001). A multivariate regression analysis revealed that pulse pressure is an independent and the most sensitive predictor for Delta%ACV/year (beta=0.389, P<0.001) among the blood pressure components. These results suggested that increase in pulse pressure promotes the progression of vascular calcification. PMID- 14757776 TI - Cytochrome P-450 inhibition attenuates hypertension induced by reductions in uterine perfusion pressure in pregnant rats. AB - The present study tested the hypothesis that cytochrome P-450 (CYP) metabolites of arachidonic acid (AA) are involved in mediating hypertension and renal vasoconstriction during chronic reductions in uterine perfusion pressure (RUPP) in pregnant rats. 1-aminobenzotriazole (ABT), a CYP enzyme inhibitor (25 mg/kg per day), or vehicle (saline 0.9%) was administered for 7 days to normal pregnant (NP) rats and to pregnant rats with chronic RUPP. RUPP rats infused with vehicle showed significantly (P<0.01) higher mean arterial pressure (MAP) (130+/-2 versus 106+/-1 mm Hg), renal vascular resistance (RVR) (22.6+/-1.8 versus 16.3+/-1.1 mm Hg/mL per minute) and lower (P<0.05) glomerular filtration rate (GFR) (1.6+/-0.1 versus 2.3+/-0.1 mL/min) than NP rats. ABT decreased (P<0.01) MAP in RUPP rats (111+/-1 mm Hg), whereas it had no effect in NP rats (108+/-2 mm Hg). CYP inhibition also attenuated the differences in renal hemodynamics observed between NP and RUPP rats. After treatment with ABT, RVR and GFR were similar in RUPP rats (19.3+/-1.5 mm Hg/mL per minute and 2.0+/-0.2 mL/min, respectively) and NP rats (16.3+/-2.4 mm Hg/mL per minute and 2.4+/-0.2 mL/min). The effects of CYP enzymes inhibitor in RUPP rats were associated with a reduction (P<0.05) of 20-HETE formation (32%) and a decreased (P<0.05) expression (33%) of CYP4A protein in renal cortex. In contrast, renal epoxygenase activity did not change in these animals. These results suggest that 20-HETE contributes to hypertension and renal vasoconstriction induced by chronic RUPP in pregnant rats. PMID- 14757777 TI - AT1 receptor blocker added to ACE inhibitor provides benefits at advanced stage of hypertensive diastolic heart failure. AB - Diastolic heart failure (DHF) has become a social burden; however, evidences leading to its therapeutic strategy are lacking. This study investigated effects of addition of angiotensin II type 1 receptor blocker (ARB) to angiotensin converting enzyme inhibitor (ACEI) at advanced stage of DHF in hypertensive rats. Dahl salt-sensitive rats fed 8% NaCl diet from age 7 weeks served as DHF model, and those fed a normal chow served as control. The DHF model rats were arbitrarily assigned to 3 treatment regimens at age 17 weeks: ACEI (temocapril 0.4 mg/kg per day), combination of ACEI (temocapril 0.2 mg/kg per day) with ARB (olmesartan 0.3 mg/kg per day), or placebo. At age 17 weeks, this model represents progressive ventricular hypertrophy and fibrosis, relaxation abnormality, and myocardial stiffening. Data were collected at age 20 weeks. As compared with the monotherapy with ACEI, the addition of ARB induced more prominent suppression of ventricular hypertrophy and fibrosis, leading to suppression of myocardial stiffening, improvement of relaxation, and inhibition of hemodynamic deterioration. Such benefits were associated with greater decreases in reactive oxygen species (ROS) generation, macrophage infiltration, and gene expression of transforming growth factor (TGF)-beta(1) and interleukin (IL)-1beta, but not with changes in gene expression of monocyte chemoattractant protein (MCP)-1 and tumor necrosis factor (TNF)-alpha. Thus, ARB added to ACEI provides more benefits as compared with ACEI alone in DHF when initiated at an advanced stage. The additive effects are likely provided through more prominent suppression of ROS generation and inflammatory changes without effects on expression of MCP-1 and TNF-alpha. PMID- 14757778 TI - Androgens are necessary for the development of fructose-induced hypertension. AB - Hyperinsulinemia and insulin resistance are closely associated with hypertension in humans and in animal models. Gender differences have been found in the development of hypertension in fructose-fed rats. The objectives of the present study were, first, to clarify whether androgens are required in the development of hyperinsulinemia, insulin resistance, and hypertension in fructose-fed rats, and second, to determine if cyclooxygenase-1 and cyclooxygenase-2 are also increased in the arteries of these rats. Male rats were gonadectomized or sham operated and fed a 60% fructose diet beginning at age 7 weeks. Blood pressure was measured by a tail-cuff method, and an oral glucose tolerance test was performed to assess insulin sensitivity after 8 weeks of fructose feeding. Cyclooxygenase-1 and cyclooxygenase-2 mRNA expression was also assessed in the thoracic aortae and mesenteric arteries. Gonadectomy prevented hypertension from developing in the fructose-fed rats, but hyperinsulinemia and insulin resistance developed. There was an increase in cyclooxygenase-2 expression in the thoracic aortae and mesenteric arteries of the fructose-fed sham-operated rats while the expression of cyclooxygenase-1 remained unchanged. Gonadectomy prevented the mRNA overexpression of vascular cyclooxygenase-2 in the fructose-fed rats. These results suggest that the presence of androgens is necessary for the development of fructose-induced hypertension. Androgens apparently act as a link between hyperinsulinemia/insulin resistance and hypertension in fructose-hypertensive rats. Furthermore, an increase in the expression of cyclooxygenase-2 is implicated in the development of hypertension. The mechanisms involved require further study. PMID- 14757779 TI - Overlap between whites and blacks in response to antihypertensive drugs. AB - On average, whites and blacks differ in their response to specific antihypertensive drugs. These differences are often highlighted in reviews and practice guidelines. However, there is wide variation in drug-associated changes in blood pressure within each race. The goal of this meta-analysis is to quantitate how often whites and blacks have similar responses to specific antihypertensive drugs. Computerized searches of MEDLINE (1983 to March 2003) and manual searches of references listed in identified articles were performed. Studies were included if they provided race-specific changes in blood pressure. Fifteen studies with a total of 9307 white subjects and 2902 black subjects were analyzed. For drug-associated changes in diastolic blood pressure, the mean difference between whites and blacks ranged from 0.6 to 3.0 mm Hg while the standard deviation within each race ranged from 5.0 to 10.1 mm Hg. The percentage of whites and blacks with similar drug-associated changes in diastolic blood pressure was 90% (95% confidence interval: 81 to 99) for diuretics, 90% (95% CI: 83 to 97) for beta-blockers, 95% (95% CI: 92 to 98) for calcium channel blockers, and 81% (95% CI: 76 to 86) for angiotensin converting enzyme inhibitors. The percentage of whites and blacks with similar drug-associated changes in systolic blood pressure ranged from 83% to 93%. In conclusion, the majority of whites and blacks have similar responses to commonly used antihypertensive drugs. Clinical decisions to use a specific drug should be based on other considerations such as efficacy in individual patients, compelling indications, and cost. PMID- 14757780 TI - Association between blood pressure and resting energy expenditure independent of body size. AB - Obesity is an important risk factor for hypertension; however, the pathway through which it raises blood pressure (BP) is poorly understood. Body size is also the primary determinant of energy expenditure, and we therefore examined the joint relationship of energy expenditure and body size to blood pressure. Resting energy expenditure (REE) was measured using respiratory gas exchange in population-based samples of 997 Nigerians and 452 African Americans. In a third sample of 118 individuals, nonresting energy expenditure (ie, physical activity) was measured in addition to REE. The univariate correlation between REE and BP ranged from 0.10 to 0.22 in the 3 samples (P<0.001). In multivariate models, adiposity, whether defined by body mass, fat mass, or leptin, was no longer associated with BP, while REE remained highly significant (P<0.001). The REE-BP association also persisted after adjustment for physical activity measured with doubly labeled water. The odds ratio for hypertension among persons in the highest quartile versus the lowest quartile of REE, after adjustment for body size, was 1.7. This relationship was not the result of hypertension among the obese, because it did not vary across the range of BMI and was the same in lean Nigerians as in obese Americans. These data suggest that metabolic processes represented by REE may mediate the effect of body size on BP. The interrelationship of REE with sympathetic tone, transmembrane ion exchange, or other metabolic processes that determine energy costs at rest could provide physiological explanations for this observation. PMID- 14757781 TI - ACE Inhibition and Bradykinin-Mediated Renal Vascular Responses: EDHF Involvement. PMID- 14757783 TI - Plasma adiponectin concentrations in women with preeclampsia. PMID- 14757785 TI - Is low-heat shock protein 70 a primary or a secondary event in the development of atherosclerosis? PMID- 14757786 TI - Improving quality and safety of telephone based delivery of care: teaching telephone consultation skills. PMID- 14757787 TI - Open disclosure: the only approach to medical error. PMID- 14757788 TI - Quality and safety in health care: plus ca change, plus ca ne reste plus la meme chose. PMID- 14757789 TI - A new structure for quality improvement reports. PMID- 14757790 TI - Defining and classifying medical error: lessons for learning. PMID- 14757791 TI - Does appropriate prescribing result in safer care? PMID- 14757792 TI - Readmission to hospital: a measure of quality or outcome? PMID- 14757793 TI - Preserving moral quality in research, audit, and quality improvement. PMID- 14757794 TI - Defining and classifying medical error: lessons for patient safety reporting systems. AB - BACKGROUND: It is important for healthcare providers to report safety related events, but little attention has been paid to how the definition and classification of events affects a hospital's ability to learn from its experience. OBJECTIVES: To examine how the definition and classification of safety related events influences key organizational routines for gathering information, allocating incentives, and analyzing event reporting data. METHODS: In semi-structured interviews, professional staff and administrators in a tertiary care teaching hospital and its pharmacy were asked to describe the existing programs designed to monitor medication safety, including the reporting systems. With a focus primarily on the pharmacy staff, interviews were audio recorded, transcribed, and analyzed using qualitative research methods. RESULTS: Eighty six interviews were conducted, including 36 in the hospital pharmacy. Examples are presented which show that: (1) the definition of an event could lead to under-reporting; (2) the classification of a medication error into alternative categories can influence the perceived incentives and disincentives for incident reporting; (3) event classification can enhance or impede organizational routines for data analysis and learning; and (4) routines that promote organizational learning within the pharmacy can reduce the flow of medication error data to the hospital. DISCUSSION: These findings from one hospital raise important practical and research questions about how reporting systems are influenced by the definition and classification of safety related events. By understanding more clearly how hospitals define and classify their experience, we may improve our capacity to learn and ultimately improve patient safety. PMID- 14757795 TI - Investigation of systems to prevent diversion of opiate drugs in general practice in the UK. AB - BACKGROUND: Statutory regulations govern the procedures that must be followed by general practitioners (GPs) in the UK to minimise the risk of diversion of prescribed opiate drugs for illicit use. However, evidence presented at the trial of Harold Shipman, a GP convicted of murdering patients with diamorphine, suggests that the regulations and monitoring of GPs' prescribing are failing. AIM: To assess the policies followed by general practices in Leicestershire and Rutland with regard to the controlled drugs regulations. METHODS: A semi structured interview was administered to a purposeful sample of lead GPs to explore how their practices applied the regulations. The controlled drugs registers and drug storage facilities in these practices were inspected. A questionnaire was sent to all the remaining practices to seek information about their application of the regulations, any concerns they had about the regulations, and any suggestions for improving them. RESULTS: Of the 142 general practices in Leicestershire, the lead GP in 14 took part in the interviews. Respondents expressed dissatisfaction with current policies including the design of controlled drug registers, and generally supported the reintroduction of an inspection scheme. Ninety (70.9%) of the 127 practices to whom the questionnaire was sent responded and, of these, 31 (34.4%) no longer held a supply of controlled drugs. Those that did hold controlled drugs indicated concern about the regulations, confusion about some aspects including the return and disposal of unused drugs, and a desire for advice and support in the implementation of the regulations. Forty two of the 59 respondents who held a supply of controlled drugs (71.2%) would welcome regular inspection. CONCLUSION: GPs are confused about the controlled drugs regulations and have little support in implementing them. The suspension of inspection schemes has reduced the amount of advice and support available to them and, in consequence, the regulations are interpreted differently in different practices. These findings are cause for concern about the risk of diversion of controlled drugs, and illustrate how patient safety systems can decay when they are not maintained. PMID- 14757796 TI - Data feedback efforts in quality improvement: lessons learned from US hospitals. AB - BACKGROUND: Data feedback is a fundamental component of quality improvement efforts, but previous studies provide mixed results on its effectiveness. This study illustrates the diversity of hospital based efforts at data feedback and highlights successful strategies and common pitfalls in designing and implementing data feedback to support performance improvement. METHODS: Open ended interviews with 45 clinical and administrative staff in eight US hospitals in 2000 concerning their perceptions about the effectiveness of data feedback in supporting performance improvement efforts were analysed. The hospitals were chosen to represent a range of sizes, geographical regions, and beta blocker improvement rates over a 3 year period. Data were organized and analyzed in NUD IST 4 using the constant comparative method of qualitative data analysis. RESULTS: Although the data feedback efforts at the hospitals were diverse, the interviews suggested that seven key themes may be important: (1) data must be perceived by physicians as valid to motivate change; (2) it takes time to develop the credibility of data within a hospital; (3) the source and timeliness of data are critical to perceived validity; (4) benchmarking improves the meaningfulness of data feedback; (5) physician leaders can enhance the effectiveness of data feedback; (6) data feedback that profiles an individual physician's practices can be effective but may be perceived as punitive; (7) data feedback must persist to sustain improved performance. Embedded in several themes was the view that the effectiveness of data feedback depends not only on the quality and timeliness of the data, but also on the organizational context in which such efforts are implemented. CONCLUSIONS: Data feedback is a complex and textured concept. Data feedback strategies that might be most effective are suggested, as well as potential pitfalls in using data to promote performance improvement. PMID- 14757797 TI - Are diagnosis specific outcome indicators based on administrative data useful in assessing quality of hospital care? AB - BACKGROUND: Hospital performance reports based on administrative data should distinguish differences in quality of care between hospitals from case mix related variation and random error effects. A study was undertaken to determine which of 12 diagnosis-outcome indicators measured across all hospitals in one state had significant risk adjusted systematic (or special cause) variation (SV) suggesting differences in quality of care. For those that did, we determined whether SV persists within hospital peer groups, whether indicator results correlate at the individual hospital level, and how many adverse outcomes would be avoided if all hospitals achieved indicator values equal to the best performing 20% of hospitals. METHODS: All patients admitted during a 12 month period to 180 acute care hospitals in Queensland, Australia with heart failure (n = 5745), acute myocardial infarction (AMI) (n = 3427), or stroke (n = 2955) were entered into the study. Outcomes comprised in-hospital deaths, long hospital stays, and 30 day readmissions. Regression models produced standardised, risk adjusted diagnosis specific outcome event ratios for each hospital. Systematic and random variation in ratio distributions for each indicator were then apportioned using hierarchical statistical models. RESULTS: Only five of 12 (42%) diagnosis-outcome indicators showed significant SV across all hospitals (long stays and same diagnosis readmissions for heart failure; in-hospital deaths and same diagnosis readmissions for AMI; and in-hospital deaths for stroke). Significant SV was only seen for two indicators within hospital peer groups (same diagnosis readmissions for heart failure in tertiary hospitals and inhospital mortality for AMI in community hospitals). Only two pairs of indicators showed significant correlation. If all hospitals emulated the best performers, at least 20% of AMI and stroke deaths, heart failure long stays, and heart failure and AMI readmissions could be avoided. CONCLUSIONS: Diagnosis-outcome indicators based on administrative data require validation as markers of significant risk adjusted SV. Validated indicators allow quantification of realisable outcome benefits if all hospitals achieved best performer levels. The overall level of quality of care within single institutions cannot be inferred from the results of one or a few indicators. PMID- 14757798 TI - The use of prescribing indicators to measure the quality of care in psychiatric inpatients. AB - OBJECTIVE: To explore the potential for using seven prescribing indicators, individually and in combination, to measure prescribing quality for hospitalised psychiatric patients. DESIGN AND SETTING: The dataset included full details of all psychotropic medication prescribed over a 24 hour period to 4192 inpatients in 49 British mental health services in 1998. RESULTS: Despite the large size of the dataset, for three of the indicators 20 services had fewer than 10 eligible patients. There was great variation between services in indicator scores. Correlations between standardised indicator scores and total score (which omitted the indicator concerned) were above 0.3 for all but one of the indicators. Cronbach's alpha was 0.73 when this outlying indicator was removed. CONCLUSIONS: There are no routinely collected prescribing data that allow for the quality of prescribing for psychiatric patients to be monitored. Six of the seven indicators measured during this census survey appear to reflect a common attribute of the services, and the analysis suggests that they might be combined to give an overall measure of service performance. There was, however, no relationship between performance on the seventh indicator and performance on the other six. This raises questions about case mix and service level factors that might influence indicator scores independent of prescriber decision making. The psychometric properties of prescribing indicators (occurrence rates, consistency over time) are unknown. PMID- 14757799 TI - Is readmission to hospital an indicator of poor process of care for patients with heart failure? AB - BACKGROUND: Controversy exists about the appropriateness of using readmission as an indicator of the quality of care. A study was undertaken to measure the validity and predictive ability of readmission in this context. METHODS: An evaluation study was performed in patients discharged alive with heart failure from three Swiss academic medical centres. Process quality indicators were derived from evidence based guidelines for the management and treatment of heart failure. Readmissions were calculated from hospital administrative data. The predictive ability of readmissions was evaluated using bivariate and multivariate analyses, and validity by calculating sensitivity, specificity, positive and negative predictive value, using process indicators as the "gold standard". RESULTS: Of 1055 eligible patients discharged alive, 139 (13.2%) were readmitted within 30 days. The adjusted odds ratio (OR) for absence of measurement of left ventricular function was 0.70 (95% CI 0.45 to 1.08) for readmissions. In patients with left ventricular systolic dysfunction, three dose categories of angiotensin converting enzyme inhibitor were examined using ordinal logistic regression. The adjusted OR for these categories was 1.07 (95% CI 0.56 to 2.06) for readmissions. When using process indicators as the gold standard to assess the validity of readmissions, sensitivity ranged from 0.08 to 0.17 and specificity from 0.86 to 0.93. CONCLUSIONS: Readmission did not predict and was not a valid indicator of the quality of care for patients with heart failure admitted to three Swiss university hospitals. PMID- 14757800 TI - Introducing criteria based audit into Ugandan maternity units. AB - PROBLEM: Maternal mortality in Uganda has remained unchanged at 500/100 000 over the past 10 years despite concerted efforts to improve the standard of maternity care. It is especially difficult to improve standards in rural areas, where there is little money for improvements. Furthermore, staff may be isolated, poorly paid, disempowered, lacking in morale, and have few skills to bring about change. DESIGN: Training programme to introduce criteria based audit into rural Uganda. SETTING: Makerere University Medical School, Mulago Hospital (large government teaching hospital in Kampala), and Mpigi District (rural area with 10 small health centres around a district hospital). STRATEGIES FOR CHANGE: Didactic teaching about criteria based audit followed by practical work in own units, with ongoing support and follow up workshops. EFFECTS OF CHANGE: Improvements were seen in many standards of care. Staff showed universal enthusiasm for the training; many staff produced simple, cost-free improvements in their standard of care. LESSONS LEARNT: Teaching of criteria based audit to those providing health care in developing countries can produce low cost improvements in the standards of care. Because the method is simple and can be used to provide improvements even without new funding, it has the potential to produce sustainable and cost effective changes in the standard of health care. Follow up is needed to prevent a waning of enthusiasm with time. PMID- 14757801 TI - Organisational trust: the keystone to patient safety. AB - Trust is an essential part of health care-not only between clinicians and patients but also between staff and management. Research shows us that trust has a beneficial impact on many aspects of working life, including job satisfaction and organisational effectiveness, and both these factors have been shown to affect the quality of patient care. In addition, trust will now be the keystone for any system developed for services to learn from untoward incidents, such as the Reporting and Learning System of the National Patient Safety Agency in the UK. This type of trust is complex and is explored in terms of what staff need from management and the potential conflicts that might be involved in developing trust in a healthcare organisation. This paper looks at the societal and emotional context of health care today and at research from other organisations which shows the factors that must be in place to establish trust. It reviews the attributes of leaders who are seen as trustworthy, and looks at how all this can be used to increase the reporting of and learning from error. PMID- 14757802 TI - The next phase of healthcare improvement: what can we learn from social movements? AB - To date, improvement in health care has relied mainly on a "top down" programme by programme approach to service change and development. This has spawned a multitude of different and often impressive improvement schemes and activities. We question whether what has been happening will be sufficient to achieve the desired scale of change within the time scales set. Is it a case of "more of the same" or are there new and different approaches that might now be usefully implemented? Evidence from the social sciences suggests that other perspectives may help to recast large scale organisational change efforts in a new light and offer a different, though complementary, approach to improvement thinking and practice. Particularly prominent is the recognition that such large scale change in organisations relies not only on the "external drivers" but on the ability to connect with and mobilise people's own "internal" energies and drivers for change, thus creating a "bottom up" locally led "grass roots" movement for improvement and change. PMID- 14757803 TI - When does quality improvement count as research? Human subject protection and theories of knowledge. AB - The publication of insights from a quality improvement project recently precipitated a ruling by the lead federal regulatory agency that regulations providing protection for human subjects of research should apply. The required research review process did not match the rapid changes, small samples, limited documentation, clinician management, and type of information commonly used in quality improvement. Yet quality improvement can risk harm to patients, so some review might be in order. The boundaries and processes are not clear. Efforts have been made to determine what constitutes "research", but this has proved difficult and often yields irrational guidance with regard to protection of patients. Society needs a workable way to separate activities that will improve care, on the one hand, and those that constitute research, on the other. Practitioners who lead both quality improvement and research projects claim that those which rapidly give feedback to the care system that generated the data, aiming to change practices within that system, are "quality improvement" no matter whether the findings are published, whether the project is grant funded, and whether contemporaneous controls do not have the intervention. This criterion has not previously been proposed as a possible demarcation. The quandaries of which projects to put through research review and how to ensure ethical implementation of quality improvement need to be resolved. PMID- 14757804 TI - Quest for quality care and patient safety: the case of Singapore. AB - Quality of care in Singapore has seen a paradigm shift from a traditional focus on structural approaches to a broader multidimensional concept which includes the monitoring of clinical indicators and medical errors. Strong political commitment and institutional capacities have been important factors for making the transition. What is still lacking, however, is a culture of rigorous programme evaluation, public involvement, and patient empowerment. Despite these imperfections, Singapore has made considerable strides and its experience may hold lessons for other small developing countries in the common quest for quality care and patient safety. PMID- 14757805 TI - Iatrogenic illness on a general medical service at a university hospital. 1981. AB - We found that 36% of 815 consecutive patients on a general medical service of a university hospital had an iatrogenic illness. In 9% of all persons admitted, the incident was considered major in that it threatened life or produced considerable disability. In 2% of the 815 patients, the iatrogenic illness was believed to contribute to the death of the patient. Exposure to drugs was a particularly important factor in determining which patients had complications. Given the increasing number and complexity of diagnostic procedures and therapeutic agents, monitoring of untoward events is essential, and attention should be paid to educational efforts to reduce the risks of iatrogenic illness. PMID- 14757806 TI - Iatrogenic illness: a call for decision support tools to reduce unnecessary variation. PMID- 14757807 TI - "Cotton Mather, you dog, dam you! I'l inoculate you with this; with a pox to you": smallpox inoculation, Boston, 1721. PMID- 14757810 TI - A mutation in the general membrane trafficking machinery and hydrocephaly. PMID- 14757811 TI - Histone H3 variants and modifications on transcribed genes. PMID- 14757812 TI - Genome annotation by high-throughput 5' RNA end determination. AB - Complete gene identification and annotation, including alternative transcripts, remains a challenge in understanding genome organization. Such annotation can be achieved by a combination of computational analysis and experimental confirmation. Here, we describe a high-throughput technique, trans-spliced exon coupled RNA end determination (TEC-RED), that identifies 5' ends of expressed genes in nematodes. TEC-RED can distinguish coding regions from regulatory regions and identify genes as well as their alternative transcripts that have different 5' ends. Application of TEC-RED to approximately 10% of the Caenorhabditis elegans genome yielded tags 75% of which experimentally verified predicted 5'-RNA ends and 25% of which provided previously unknown information about 5'-RNA ends, including the identification of 99 previously unknown genes and 32 previously unknown operons. This technique will be applicable in any organisms that have a trans-splicing reaction from spliced leader RNA. We also describe an efficient sequential method for concatenating short sequence tags for any serial analysis of gene expression-like techniques. PMID- 14757813 TI - Signal transduction pathways that inhibit hepatitis B virus replication. AB - The replication of hepatitis B virus (HBV) in hepatocytes is strongly inhibited in response to IFN-alpha/beta and IFN-gamma. Although it has been previously demonstrated that IFN-alpha/beta eliminates HBV RNA-containing capsids from the cell in a proteasome-dependent manner, the precise cellular pathway that mediates this antiviral effect has not been identified. Because IFN-induced signal transduction involves kinase-mediated activation of gene expression, we used an immortalized hepatocyte cell line that replicates HBV in an IFN-sensitive manner to investigate the role of cellular kinase activity and the cellular transcription and translation machinery in the antiviral effect. Our results indicate that Janus kinase activity is required for the antiviral effect of IFN against HBV, but that phosphatidylinositol 3-kinase, cyclin-dependent kinase, mitogen-activated protein kinase, and NF-kappaB activity are not. Additionally, we found that inhibitors of cellular transcription and translation completely abolish the antiviral effect, which also appears to require cellular kinase activity downstream of signal transduction and gene expression. Collectively, these results identify IFN-regulated pathways that interrupt the HBV replication cycle by eliminating viral RNA-containing capsids from the cell, and they provide direction for discovery of the terminal effector molecules that ultimately mediate this antiviral effect. PMID- 14757814 TI - Evidence that vaccinia virulence factor E3L binds to Z-DNA in vivo: Implications for development of a therapy for poxvirus infection. AB - The E3L gene product found in all poxviruses is required for the lethality of mice in vaccinia virus infection. Both the C-terminal region, consisting of a double-stranded RNA-binding motif, and the N-terminal region (vZ(E3L)), which is similar to the Zalpha family of Z-DNA-binding proteins, are required for infection. It has recently been demonstrated that the function of the N-terminal domain depends on its ability to bind Z-DNA; Z-DNA-binding domains from unrelated mammalian proteins fully complement an N-terminal deletion of E3L. Mutations that decrease affinity for Z-DNA have similar effects in decreasing pathogenicity. Compounds that block the Z-DNA-binding activity of E3L may also limit infection by the poxvirus. Here we show both an in vitro and an in vivo assay with the potential to be used in screening for such compounds. Using a conformation specific yeast one-hybrid assay, we compared the results for Z-DNA binding of vZ(E3L) with those for human Zbeta(ADAR1), a peptide that has similarity to the Zalpha motif but does not bind Z-DNA, and with a mutant of hZbeta(ADAR1), which binds Z-DNA. The results suggest that this system can be used for high-throughput screening. PMID- 14757815 TI - Elucidation of signaling properties of vasopressin receptor-related receptor 1 by using the chimeric receptor approach. AB - The identification of endogenous or surrogate ligands for orphan G protein coupled receptors (GPCRs) represents one of the most important tasks in GPCR biology and pharmacology. The challenge lies in choosing an appropriate assay in the absence of ways to activate the receptor of interest. We investigated the signaling pathway for an orphan GPCR referred to here as vasopressin receptor related receptor 1 (VRR1) by generating a chimeric receptor, V1a/VRR1. The engineered construct contained vasopressin V1a receptor with all three intracellular loops and C terminus replaced by those of VRR1. The chimera behaved like a typical GPCR when transiently and stably expressed in mammalian cell lines based on radioligand binding and receptor internalization studies. Upon arginine vasopressin stimulation, this chimeric receptor induced robust calcium mobilization and increase of adenylate cyclase activity. The observed signaling activities are through the activation of the chimera instead of endogenously expressed receptors, as single amino acid changes in the second transmembrane regions of the chimera drastically reduced receptor efficacy and potency. Our results suggest that VRR1 has dual signaling properties in coupling to both G(q) and G(S) pathways. Analysis of native VRR1 receptor signaling pathway by using a recently identified ligand for VRR1 confirmed this conclusion and therefore validated the utility of the chimeric receptor approach for signaling pathway identification. PMID- 14757816 TI - Freeze-frame inhibitor captures acetylcholinesterase in a unique conformation. AB - The 1,3-dipolar cycloaddition reaction between unactivated azides and acetylenes proceeds exceedingly slowly at room temperature. However, considerable rate acceleration is observed when this reaction occurs inside the active center gorge of acetylcholinesterase (AChE) between certain azide and acetylene reactants, attached via methylene chains to specific inhibitor moieties selective for the active center and peripheral site of the enzyme. AChE catalyzes the formation of its own inhibitor in a highly selective fashion: only a single syn1-triazole regioisomer with defined substitution positions and linker distances is generated from a series of reagent combinations. Inhibition measurements revealed this syn1 triazole isomer to be the highest affinity reversible organic inhibitor of AChE with association rate constants near the diffusion limit. The corresponding anti1 isomer, not formed by the enzyme, proved to be a respectable but weaker inhibitor. The crystal structures of the syn1- and anti1-mouse AChE complexes at 2.45- to 2.65-A resolution reveal not only substantial binding contributions from the triazole moieties, but also that binding of the syn1 isomer induces large and unprecedented enzyme conformational changes not observed in the anti1 complex nor predicted from structures of the apoenzyme and complexes with the precursor reactants. Hence, the freeze-frame reaction offers both a strategically original approach for drug discovery and a means for kinetically controlled capture, as a high-affinity complex between the enzyme and its self-created inhibitor, of a highly reactive minor abundance conformer of a fluctuating protein template. PMID- 14757818 TI - Human endogenous retrovirus K solo-LTR formation and insertional polymorphisms: implications for human and viral evolution. AB - Human endogenous retroviruses (HERVs) are a potential source of genetic diversity in the human genome. Although many of these elements have been inactivated over time by the accumulation of deleterious mutations or internal recombination leading to solo-LTR formation, several members of the HERV-K family have been identified that remain nearly intact and probably represent recent integration events. To determine whether HERV-K elements have caused recent changes in the human genome, we have undertaken a study of the level of HERV-K polymorphism that exists in the human population. By using a high-resolution unblotting technique, we analyzed 13 human-specific HERV-K elements in 18 individuals. We found that solo LTRs have formed at five of these loci. These results enable the estimation of HERV solo-LTR formation in the human genome and indicate that these events occur much more frequently than described in inbred mice. Detailed sequence analysis of one provirus shows that solo-LTR formation occurred at least three separate times in recent history. An unoccupied preintegration site also was present at this locus in two individuals, indicating that although the age of this provirus is estimated to be approximately 1.2 million years, it has not yet become fixed in the human population. PMID- 14757817 TI - Major events in the genome evolution of vertebrates: paranome age and size differ considerably between ray-finned fishes and land vertebrates. AB - It has been suggested that fish have more genes than humans. Whether most of these additional genes originated through a complete (fish-specific) genome duplication or through many lineage-specific tandem gene or smaller block duplications and family expansions continues to be debated. We analyzed the complete genome of the pufferfish Takifugu rubripes (Fugu) and compared it with the paranome of humans. We show that most paralogous genes of Fugu are the result of three complete genome duplications. Both relative and absolute dating of the complete predicted set of protein-coding genes suggest that initial genome duplications, estimated to have occurred at least 600 million years ago, shaped the genome of all vertebrates. In addition, analysis of >150 block duplications in the Fugu genome clearly supports a fish-specific genome duplication (approximately equal to 320 million years ago) that coincided with the vast radiation of most modern ray-finned fishes. Unlike the human genome, Fugu contains very few recently duplicated genes; hence, many human genes are much younger than fish genes. This lack of recent gene duplication, or, alternatively, the accelerated rate of gene loss, is possibly one reason for the drastic reduction of the genome size of Fugu observed during the past 100 million years or so, subsequent to the additional genome duplication that ray-finned fishes but not land vertebrates experienced. PMID- 14757819 TI - Production of fertile offspring from genetically infertile male mice. AB - A number of recessive autosomal genes cause male infertility. Male mice homozygous for the blind-sterile (bs/bs) and quaking-sterile (qk/qk) gene mutations are sterile, because they either do not produce any spermatozoa or produce only a few abnormal spermatozoa. Mice lacking the cyclic AMP responsive element modulator gene are sterile due to failure of spermiogenesis. All these mice, however, are able to produce fertile offspring when their spermatozoa or round spermatids are injected into oocytes of normal females. This implies that genetic and epigenetic elements necessary for syngamy and embryonic development are established in round spermatids and spermatozoa of these animals, even though their spermatogenic cells are destined to die (bs/bs and qk/qk) or are programmed to undergo apoptosis (cyclic AMP responsive-element modulator-null) without becoming functional spermatozoa. PMID- 14757820 TI - Tracking evolving communities in large linked networks. AB - We are interested in tracking changes in large-scale data by periodically creating an agglomerative clustering and examining the evolution of clusters (communities) over time. We examine a large real-world data set: the NEC CiteSeer database, a linked network of >250,000 papers. Tracking changes over time requires a clustering algorithm that produces clusters stable under small perturbations of the input data. However, small perturbations of the CiteSeer data lead to significant changes to most of the clusters. One reason for this is that the order in which papers within communities are combined is somewhat arbitrary. However, certain subsets of papers, called natural communities, correspond to real structure in the CiteSeer database and thus appear in any clustering. By identifying the subset of clusters that remain stable under multiple clustering runs, we get the set of natural communities that we can track over time. We demonstrate that such natural communities allow us to identify emerging communities and track temporal changes in the underlying structure of our network data. PMID- 14757821 TI - A method for finding communities of related genes. AB - We present a method for creating a network of gene co-occurrences from the literature and partitioning it into communities of related genes. The way in which our method identifies communities makes it likely that the component genes of each community will be related by their function. The method processes a large database of article abstracts, synthesizing information from many sources to shed light on groups of genes that have been shown to interact. It is a tool to be used by researchers in the biomedical sciences to swiftly search for known interactions and to provide insight into unexplored connections. The partitioning procedure is designed to be particularly applicable to large networks in which individual nodes may play a role in more than one community. In this paper, we explain the details of the method, in particular the partitioning process. We also apply the method to produce communities of genes related to colon cancer and show that the results are useful. PMID- 14757822 TI - Linking histone deacetylation with the repair of DNA breaks. PMID- 14757823 TI - Crystal and molecular structure of a benzo[a]pyrene 7,8-diol 9,10-epoxide N2 deoxyguanosine adduct: absolute configuration and conformation. AB - Benzo[a]pyrene 7,8-diol 9,10-epoxide adducts in DNA are implicated in mutagenesis, and their formation from the diol epoxides and subsequent incorrect replication by human DNA polymerases provide an attractive mechanism for the induction of cancer by this highly carcinogenic hydrocarbon and its diol epoxide metabolites. Here, we describe the crystal structure of such an adduct at the exocyclic amino group of a purine nucleoside. The present adduct derives from trans opening at C10 of the (-)-(7S,8R)-diol (9R,10S)-epoxide enantiomer by the exocyclic N(2)-amino group of deoxyguanosine. In the crystal, the pyrene rings of adjacent molecules stack with each other, but the guanine bases do not stack either intermolecularly with each other or intramolecularly with the pyrene. The most notable features of the molecular structure are (i) independent and unambiguous proof of the absolute configuration of the adduct based on the spatial relationship between the known chiral carbon atoms of the deoxyribose and the four asymmetric centers in the hydrocarbon moiety; (ii) visualization of the relative orientations of the pyrene and guanine ring systems as well as the conformation of the partially saturated hydrocarbon ring (comprising carbon atoms 7, 8, 9, and 10), both of which conformational features in the crystal are in good agreement with deductions from NMR and CD measurements in solution; and (iii) the presence in the crystal of a syn glycosidic torsion angle, a conformation that is unusual in B-DNA but that may be involved in error-prone replication of these benzo[a]pyrene 7,8-diol 9,10-epoxide deoxyguanosine adducts by DNA polymerases. PMID- 14757824 TI - The metabolic world of Escherichia coli is not small. AB - To elucidate the organizational and evolutionary principles of the metabolism of living organisms, recent studies have addressed the graph-theoretic analysis of large biochemical networks responsible for the synthesis and degradation of cellular building blocks [Jeong, H., Tombor, B., Albert, R., Oltvai, Z. N. & Barabasi, A. L. (2000) Nature 407, 651-654; Wagner, A. & Fell, D. A. (2001) Proc. R. Soc. London Ser. B 268, 1803-1810; and Ma, H.-W. & Zeng, A.-P. (2003) Bioinformatics 19, 270-277]. In such studies, the global properties of the network are computed by considering enzymatic reactions as links between metabolites. However, the pathways computed in this manner do not conserve their structural moieties and therefore do not correspond to biochemical pathways on the traditional metabolic map. In this work, we reassessed earlier results by digitizing carbon atomic traces in metabolic reactions annotated for Escherichia coli. Our analysis revealed that the average path length of its metabolism is much longer than previously thought and that the metabolic world of this organism is not small in terms of biosynthesis and degradation. PMID- 14757825 TI - Rapid modulation of osteoblast ion channel responses by 1alpha,25(OH)2-vitamin D3 requires the presence of a functional vitamin D nuclear receptor. AB - 1alpha,25(OH)(2)-Vitamin D(3) (1,25D) modulates osteoblast gene expression of bone matrix proteins via a nuclear vitamin D receptor (VDR) and also modifies the electrical state of the plasma membrane through rapid nongenomic mechanisms still not fully understood. The physiological significance of 1,25D membrane-initiated effects remains unclear. To elucidate whether the VDR is required for 1,25D promoted electrical responses, we studied 1,25D modulation of ion channel activities in calvarial osteoblasts isolated from VDR knockout (KO) and WT mice. At depolarizing potentials, Cl(-) currents were significantly potentiated (13.5 +/- 1.6-fold increase, n = 12) by 5 nM 1,25D in VDR WT but not in KO (0.96 +/- 0.3 fold increase, n = 11) osteoblasts. L-type Ca(2+) currents significantly shift their peak activation by -9.3 +/- 0.7 mV (n = 10) in the presence of 5 nM 1,25D in VDR WT but not in KO cells, thus facilitating Ca(2+) influx. Furthermore, we found that 1,25D significantly increased whole-cell capacitance in VDR WT (DeltaCap = 2.3 +/- 0.4 pF, n = 8) but not in KO osteoblasts (DeltaCap = 0.3 +/- 0.1 pF, n = 8); this corresponds to a rapid (1-2 min) fusion in WT of 71 +/- 33 versus in KO only 9 +/- 6 individual secretory granules. We conclude that, in calvarial osteoblasts, 1,25D modulates ion channel activities only in cells with a functional VDR and that this effect is coupled to exocytosis. This is a demonstration of the requirement of a functional classic steroid receptor for the rapid hormonal modulation of electric currents linked to secretory activities in a target cell. PMID- 14757826 TI - The endogenous retroviral locus ERVWE1 is a bona fide gene involved in hominoid placental physiology. AB - The definitive demonstration of a role for a recently acquired gene is a difficult task, requiring exhaustive genetic investigations and functional analysis. The situation is indeed much more complicated when facing multicopy gene families, because most or portions of the gene are conserved among the hundred copies of the family. This is the case for the ERVWE1 locus of the human endogenous retrovirus W family (HERV-W), which encodes an envelope glycoprotein (syncytin) likely involved in trophoblast differentiation. Here we describe, in 155 individuals, the positional conservation of this locus and the preservation of the envelope ORF. Sequencing of the critical elements of the ERVWE1 provirus showed a striking conservation among the 48 alleles of 24 individuals, including the LTR elements involved in the transcriptional machinery, the splice sites involved in the maturation of subgenomic Env mRNA, and the Env ORF. The functionality and tissue specificity of the 5' LTR were demonstrated, as well as the fusogenic activity of the envelope polymorphic variants. Such functions were also shown to be preserved in the orthologous loci isolated from chimpanzee, gorilla, orangutan, and gibbon. This functional preservation among humans and during evolution strongly argued for the involvement of this recently acquired retroviral envelope glycoprotein in hominoid placental physiology. PMID- 14757827 TI - Single-nucleotide promoter polymorphism alters transcription of neuronal nitric oxide synthase exon 1c in infantile hypertrophic pyloric stenosis. AB - Infantile hypertrophic pyloric stenosis (IHPS), characterized by enlarged pyloric musculature and gastric-outlet obstruction, is associated with altered expression of neuronal nitric oxide synthase (nNOS). Here we have studied molecular mechanisms by which nNOS gene expression is altered in pyloric tissues of 16 infants with IHPS and 9 controls. A significant decreased expression of total nNOS mRNA was found by quantitative RT-PCR in IHPS after normalization against GAPDH, which predominantly affected exon 1c with a reduction of 88% compared with controls (P < 0.001). After normalization against the neuronal-specific gene PGP9.5, expression of exon 1c was still decreased (P < 0.001), whereas expression of exon 1f was increased significantly (P = 0.001), indicating a compensatory up regulation of this nNOS mRNA variant. DNA samples of 16 IHPS patients and 81 controls were analyzed for nNOS exon 1c promoter mutations and single-nucleotide polymorphism (SNP). Sequencing of the 5'-flanking region of exon 1c revealed mutations in 3 of 16 IHPS tissues, whereas 81 controls showed the wild-type sequence exclusively. Carriers of the A allele of a previously uncharacterized nNOS exon 1c promoter SNP (-84G --> A) had increased risk for development of IHPS (odds ratio, 8.0; 95% confidence interval, 2.5-25.6). Reporter gene assays revealed an unchanged promoter activity for mutations but a approximately 30% decrease for the -84A SNP (P < 0.001). In summary, our findings indicate that genetic alterations in the nNOS exon 1c regulatory region influence expression of the nNOS gene and may contribute to the pathogenesis of IHPS. PMID- 14757828 TI - Heterodimerization of V1a and V2 vasopressin receptors determines the interaction with beta-arrestin and their trafficking patterns. AB - V1a vasopressin receptor (V1aR) and V2 vasopressin receptor (V2R) present distinct mechanisms of agonist-promoted trafficking. Although both receptors are endocytosed by way of beta-arrestin-dependent processes, beta-arrestin dissociates rapidly from V1aR, allowing its rapid recycling to the plasma membrane while beta-arrestin remains associated with V2R in the endosomes, leading to their intracellular accumulation. Here, we demonstrate that, when coexpressed, the two receptors can be endocytosed as stable heterodimers. On activation with a nonselective agonist, both receptors cotrafficked with beta arrestin in endosomes where the stable interaction inhibited the recycling of V1aR to the plasma membrane, thus conferring a V2R-like endocytotic/recycling pattern to the V1aR/V2R heterodimer. Coexpression of the constitutively internalized R137HV2R mutant with V1aR was sufficient to promote cointernalization of V1aR in beta-arrestin-positive vesicles even in the absence of agonist stimulation. This finding indicates that internalization of the heterodimer does not require activation of each of the protomers. Consistent with this notion, a V1aR-selective agonist led to the coendocytosis of V2R. In that case, however, the V1aR/V2R heterodimer was not stably associated with beta arrestin, and both receptors were recycled back to the cell surface, indicating that the complex followed the V1aR endocytotic/recycling path. Taken together, these results suggest that heterodimerization regulates the endocytotic processing of G protein-coupled receptors and that the identity of the activated protomer within the heterodimer determines the fate of the internalized receptors. PMID- 14757829 TI - Constitutive and inducible trypsin proteinase inhibitor production incurs large fitness costs in Nicotiana attenuata. AB - Plant trypsin proteinase inhibitors (TPIs) are potent herbivore- and jasmonate (JA)-induced defenses, but support for the commonly invoked explanation for their inducible expression, namely their associated fitness costs, has been elusive. To determine whether the expression of TPIs incurs fitness costs, we expressed 175 bp of the seven-domain pi from Nicotiana attenuata in an antisense orientation in a TPI-producing genotype (WT) of N. attenuata to reduce TPI expression. Moreover, we expressed the full-length seven-domain pi in a sense orientation under control of a constitutive promoter to restore TPI activity in a natural genotype unable to produce TPIs because of a mutation in its endogenous pi gene. Lifetime reproductive output was determined from high and low TPI-producing plants of the same genetic background with and without JA elicitation and grown in the same pot to simulate natural competitive and nutrient regimes. Transformants with either low or no TPI activity grew faster and taller, flowered earlier, and produced more seed capsules (25-53%) than did neighboring TPI-producing genotypes, and JA elicitation increased TPI production and decreased seed capsule production further. Growth under high light levels only marginally reduced these fitness costs. Results were similar regardless of whether TPI activity was suppressed or restored by transformation: the larger the difference in TPI activity between neighbors, the larger the difference in seed capsule production (R(2) = 0.57). TPI production is costly for a plant's components of fitness when grown under realistic competitive regimes and is consistent with the hypothesis that inducibility evolved as a cost-saving mechanism. PMID- 14757830 TI - Ca2+/calmodulin transfers the membrane-proximal lipid-binding domain of the v SNARE synaptobrevin from cis to trans bilayers. AB - Soluble N-ethylmaleimide-sensitive fusion protein attachment protein receptor (SNARE) protein interactions at the synaptic vesicle/plasma membrane interface play an essential role in neurotransmitter release. The membrane-proximal region (amino acids 77-90) of the v-SNARE vesicle-associated membrane protein 2 (VAMP 2, synaptobrevin) binds acidic phospholipids or Ca(2+)/calmodulin in a mutually exclusive manner, processes that are required for Ca(2+)-dependent exocytosis. To address the mechanisms involved, we asked whether this region of VAMP can interact with cis (outer vesicle leaflet) and/or trans (inner plasma membrane leaflet) lipids. To evaluate cis lipid binding, recombinant VAMP was reconstituted into liposomes and accessibility to site-directed antibodies was probed by surface plasmon resonance. Data indicated that the membrane-proximal domain of VAMP dips into the cis lipid bilayer, sequestering epitopes between the tetanus toxin cleavage site and the membrane anchor. These epitopes were unmasked by VAMP double mutation W89A, W90A, which abolishes lipid interactions. To evaluate trans lipid binding, VAMP was reconstituted in cis liposomes, which were then immobilized on beads. The ability of VAMP to capture protein-free (3)H labeled trans liposomes was then measured. When cis lipid interactions were eliminated by omitting negatively charged lipids, trans lipid binding to VAMP was revealed. In contrast, when cis and trans liposomes both contained acidic headgroups (i.e., approximating physiological conditions), cis lipid interactions totally occluded trans lipid binding. In these conditions Ca(2+)/calmodulin displaced cis inhibition, transferring the lipid-binding domain of VAMP from the cis to the trans bilayer. Our results suggest that calmodulin acts as a unidirectional Ca(2+)-activated shuttle that docks the juxtamembrane portion of the v-SNARE in the target membrane to prepare fusion. PMID- 14757831 TI - Interpopulational differences in progesterone levels during conception and implantation in humans. AB - Clinical studies of women from the United States demonstrate a sensitivity of the ovarian system to energetic stress. Even moderate exercise or caloric restriction can lead to lower progesterone levels and failure to ovulate. Yet women in many nonindustrial populations experience as many as a dozen pregnancies in a lifetime despite poor nutritional resources, heavy workloads, and typical progesterone levels only about two-thirds of those of U.S. women. Previous cross-sectional studies of progesterone may, however, suffer from inadvertent selection bias. In a noncontracepting population, the most fecund women, who might be expected to have the highest progesterone, are more likely to be pregnant or breastfeeding and hence unavailable for a cross-sectional study of the ovarian cycle. The present longitudinal study was designed to ascertain whether lower progesterone also characterizes conception, implantation, and gestation in women from nonindustrialized populations. We compared rural Bolivian Aymara women (n = 191) to women from Chicago (n = 29) and found that mean-peak-luteal progesterone in the ovulatory cycles of Bolivian women averaged approximately 71% that of the women from Chicago. In conception cycles, progesterone levels in Bolivian women during the periovulatory period were approximately 63%, and during the peri implantation period were approximately 50%, those of the U.S. women. These observations argue that lower progesterone levels typically characterize the reproductive process in Bolivian women and perhaps others from nonindustrialized populations. We discuss the possible proximate and evolutionary explanations for this variation and note the implications for developing suitable hormonal contraceptives and elucidating the etiology of cancers of the breast and reproductive tract. PMID- 14757832 TI - Poly(ADP-ribose) polymerase (PARP-1) and p53 independently function in regulating double-strand break repair in primate cells. AB - PARP-1 is rapidly activated by DNA strand breaks, which finally leads to the modulation of multiple protein activities in DNA replication, DNA repair and checkpoint control. PARP-1 may be involved in homologous recombination, and poly(ADP-ribosyl)ation of p53 represents one possible mechanism that activates p53 as a recombination surveillance factor. Here, we examined the influence of PARP-1 on homology-directed double-strand break (DSB) repair by use of a fluorescence- and I-SceI- meganuclease-based assay with either episomal or chromosomally integrated DNA substrates. Surprisingly, the transient expression of both full-length PARP-1 and of a dominant negative mutant, retaining the DNA binding but lacking the catalytic domain, down-regulated DSB repair in a dose dependent manner. This effect was seen regardless of p53 status, however, with enhanced inhibition in the presence of wild-type p53. Taken together, our data reveal that PARP-1 overexpression counteracts DSB repair independently of its enzymatic activity and of poly(ADP-ribosyl)ation of p53 in particular, but synergizes with p53 in suppressing chromosomal rearrangements. PMID- 14757833 TI - Guidelines for incorporating non-perfectly matched oligonucleotides into target specific hybridization probes for a DNA microarray. AB - Sequence-specific oligonucleotide probes play a crucial role in hybridization techniques including PCR, DNA microarray and RNA interference. Once the entire genome becomes the search space for target genes/genomic sequences, however, cross-hybridization to non-target sequences becomes a problem. Large gene families with significant similarity among family members, such as the P450s, are particularly problematic. Additionally, accurate single nucleotide polymorphism (SNP) detection depends on probes that can distinguish between nearly identical sequences. Conventional oligonucleotide probes that are perfectly matched to target genes/genomic sequences are often unsuitable in such cases. Carefully designed mismatches can be used to decrease cross-hybridization potential, but implementing all possible mismatch probes is impractical. Our study provides guidelines for designing non-perfectly matched DNA probes to target DNA sequences as desired throughout the genome. These guidelines are based on the analysis of hybridization data between perfectly matched and non-perfectly matched DNA sequences (single-point or double-point mutated) calculated in silico. Large changes in hybridization temperature predicted by these guidelines for non matched oligonucleotides fit independent experimental data very well. Applying the guidelines to find oligonucleotide microarray probes for P450 genes, we confirmed the ability of our point mutation method to differentiate the individual genes in terms of thermodynamic calculations of hybridization and sequence similarity. PMID- 14757834 TI - The Trypanosoma brucei spliced leader RNA and rRNA gene promoters have interchangeable TbSNAP50-binding elements. AB - In the protist parasite Trypanosoma brucei, the small nuclear spliced leader (SL) RNA and the large rRNAs are key molecules for mRNA maturation and protein synthesis, respectively. The SL RNA gene (SLRNA) promoter recruits RNA polymerase II and consists of a bipartite upstream sequence element (USE) and an element close to the transcription initiation site. Here, we analyzed the distal part of the ribosomal (RRNA) promoter and identified two sequence blocks which, in reverse orientation, closely resemble the SLRNA USE by both sequence and spacing. A detailed mutational analysis revealed that the ribosomal (r)USE is essential for efficient RRNA transcription in vivo and that it functions in an orientation dependent manner. Moreover, we showed that USE and rUSE are functionally interchangeable and that rUSE stably interacted with an essential factor of SLRNA transcription. Finally, we demonstrated that the T.brucei homolog of the recently characterized transcription factor p57 of the related organism Leptomonas seymouri specifically bound to USE and rUSE. Since p57 and its T.brucei counterpart are homologous to SNAP50, a component of the human small nuclear RNA gene activation protein complex (SNAPc), both SLRNA and RRNA transcription in T.brucei may depend on a SNAPc-like transcription factor. PMID- 14757835 TI - A novel concept for ligand attachment to oligonucleotides via a 2'-succinyl linker. AB - Conjugation of ligands to antisense oligonucleotides is a promising approach for enhancing their effects. In this report, a new method for synthesizing oligonucleotide conjugates is described. 2'-Amino-2'-deoxy-5'-dimethoxytrityl uridine was select ively acylated with a succinic acid linker at the 2' position. This compound was incorporated at the 3' end of an oligonucleotide corresponding to the sequence of Oblimersen. The carboxyl group was protected for oligonucleotide synthesis as a benzyl ester, which could be selectively cleaved at the solid phase by a catalytic phase transfer reaction using palladium nanoparticles as catalyst. An oligonucleotide-fluorescein conjugate was prepared by condensation of aminofluorescein. Circular dichroism spectroscopic experiments showed a B-DNA type structure. The melting temperature of the duplex was only slightly lower than that of Oblimersen. Biological activity measured by western blotting resulted in a Bcl-2 target downregulation nearly identical to that of control Oblimersen on human melanoma cells, proving that this method is attractive for the binding of ligands located in the minor groove. PMID- 14757836 TI - A new gamma-interferon-inducible promoter and splice variants of an anti angiogenic human tRNA synthetase. AB - Two forms of human tryptophanyl-tRNA synthetase (TrpRS) are produced in vivo through alternative mRNA splicing. The two forms, full-length TrpRS and mini TrpRS, are catalytically active, but are distinguished by the striking anti proliferative and anti-angiogenic activity specific to mini TrpRS. Here we describe two new splice variants of human TrpRS mRNA. Their production was strongly regulated by gamma-interferon (IFN-gamma), an anti-proliferative cytokine known to stimulate the expression of other anti-angiogenic factors. A new IFN-gamma-sensitive promoter was demonstrated to drive production of these splice variants. In human endothelial cells, both the newly discovered and a previously reported promoter were shown to respond specifically to IFN-gamma and not to other cytokines such as tumor necrosis factor-alpha, transforming growth factor-beta, interleukin-4 or erythropoietin. In addition, both promoters were stimulated by the 'downstream' interferon regulatory factor 1 that, in turn, is known to be regulated by the 'upstream' signal transducer and activator of transcription 1alpha subunit. Thus, the tandem promoters provide a dual system to regulate expression and alternative splicing of human TrpRS in vivo. PMID- 14757837 TI - PCR amplification of DNA containing non-standard base pairs by variants of reverse transcriptase from Human Immunodeficiency Virus-1. AB - As the next step towards generating a synthetic biology from artificial genetic information systems, we have examined variants of HIV reverse transcriptase (RT) for their ability to synthesize duplex DNA incorporating the non-standard base pair between 2,4-diaminopyrimidine (pyDAD), a pyrimidine presenting a hydrogen bond 'donor-acceptor-donor' pattern to the complementary base, and xanthine (puADA), a purine presenting a hydrogen bond 'acceptor-donor-acceptor' pattern. This base pair fits the Watson-Crick geometry, but is joined by a pattern of hydrogen bond donor and acceptor groups different from those joining the GC and AT pairs. A variant of HIV-RT where Tyr 188 is replaced by Leu, has emerged from experiments where HIV was challenged to grow in the presence of drugs targeted against the RT, such as L-697639, TIBO and nevirapine. These drugs bind at a site near, but not in, the active site. This variant accepts the pyDAD-puADA base pair significantly better than wild type HIV-RT, and we used this as a starting point. A second mutation, E478Q, was introduced into the Y188L variant, in the event that the residual nuclease activity observed is due to the RT, and not a contaminant. The doubly mutated RT incorporated the non-standard pair with sufficient fidelity that the variant could be used to amplify oligonucleotides containing pyDAD and puADA through several rounds of a polymerase chain reaction (PCR) without losing the non-standard base pair. This is the first time where DNA containing non-standard base pairs with alternative hydrogen bonding patterns has been amplified by a full PCR. This work also illustrates a research strategy that combines in clinico pre-evolution of proteins followed by rational design to obtain an enzyme that meets a particular technological specification. PMID- 14757838 TI - Fission yeast Arp6 is required for telomere silencing, but functions independently of Swi6. AB - The actin-related proteins (Arps), which are subdivided into at least eight subfamilies, are conserved from yeast to humans. A member of the Arp6 subfamily in Drosophila, Arp4/Arp6, co-localizes with heterochromatin protein 1 (HP1) in pericentric heterochromatin. Fission yeast Schizosaccharomyces pombe possesses both an HP1 homolog and an Arp6 homolog. However, the function of S.pombe Arp6 has not been characterized yet. We found that deletion of arp6(+) impaired telomere silencing, but did not affect centromere silencing. Chromatin immunoprecipitation assays revealed that Arp6 bound to the telomere region. However, unlike Drosophila Arp4/Arp6, S.pombe Arp6 was distributed throughout nuclei. The binding of Arp6 to telomere DNA was not affected by deletion of swi6(+). Moreover, the binding of Swi6 to telomere ends was not affected by deletion of arp6(+). These results suggest that Arp6 and Swi6 function independently at telomere ends. We propose that the Arp6-mediated repression mechanism works side by side with Swi6-based telomere silencing in S.pombe. PMID- 14757839 TI - Structure of a palindromic amplicon junction implicates microhomology-mediated end joining as a mechanism of sister chromatid fusion during gene amplification. AB - Amplification of the copy number of oncogenes is frequently associated with tumor progression. Often, the amplified DNA consists of large (tens to hundreds of kilobases) 'head-to-head' inverted repeat palindromes (amplicons). Several mechanisms have been proposed to explain palindrome formation but their relative contributions in nature have been difficult to assess without precise knowledge of the sequences involved at the junction of natural amplicons. Here, we have sequenced one such junction and compared this sequence to the un-rearranged structure, allowing us to pinpoint the site of sister chromatid fusion. Our results support a novel model, consistent with all described sister chromatid fusions, in which sister chromatid fusion is initiated by microhomology-mediated end joining of double strand breaks. PMID- 14757840 TI - MDM2 and prognosis. AB - The cellular stress response pathway regulated by the p53 tumor suppressor is critical to the maintenance of genomic integrity and to the prevention of oncogenic transformation. Intracellular levels of p53 are tightly regulated by an autoregulatory feedback loop comprised of p53 and MDM2. It might be predicted that disruption of this loop, either through p53 mutation or overexpression of MDM2, would be a negative prognostic marker for cancer development, likelihood of relapse, or response to therapy. In fact, although MDM2 overexpression is common in cancer, it can be both a positive and a negative predictor of outcome in different tumors, and its significance as a biomarker remains controversial. Data from a number of different tumor types are reviewed for the predictive significance of MDM2 expression, along with evidence for different mechanisms of MDM2 overexpression in these different tumors. In light of the biological complexities underlying the p53-MDM2 loop, it is, perhaps, not surprising that no simple paradigm exists that is generally applicable. Much work remains to be done to elucidate the basic mechanisms underlying the physical interactions between the two proteins, the role of protein modifications in altering those interactions, and also the genetic and transcriptional deregulations by which protein levels are altered in human cancers. Only in this way will truly biologically relevant predictive factors emerge. PMID- 14757841 TI - Mdm2 in the response to radiation. AB - Murine double minute 2 (Mdm2) is a critical component of the responses to both ionizing and UV radiation. The level of Mdm2 expression determines the extent to which radiation induces an increase in the activity of the p53 tumor suppressor. Mdm2 acts as a survival factor in many cell types by limiting the apoptotic function of p53. In addition, expression of mdm2 is induced in response to DNA damage, and the resulting high levels of Mdm2 protein are thought to shorten the length of the cell cycle arrest established by p53 in the radiation response. Increased levels of Mdm2 appear to ensure that the activity of p53 returns to its low basal levels in surviving cells. Decreased levels of Mdm2 sensitize cells to ionizing radiation. Thus, Mdm2 is a potential target for therapeutic intervention because its inhibition may radiosensitize the subset of human tumors expressing wild-type p53 such that radiotherapy is more efficacious. PMID- 14757842 TI - Inhibition of the p53-MDM2 interaction: targeting a protein-protein interface. AB - MDM2 inhibits p53 transcriptional activity, favors its nuclear export, and stimulates its degradation. Inhibition of the p53-MDM2 interaction with synthetic molecules should therefore lead to both the nuclear accumulation and the activation of p53 followed by the death of the tumor cells from apoptosis. Inhibitors of the p53-MDM2 interaction might be attractive new anticancer agents that could be used to activate wild-type p53 in tumors. This review describes our current knowledge on the properties of the existing p53-MDM2 antagonists. Because the discovery of modulators of protein-protein interactions is an emerging field in drug discovery, the strategy used for designing inhibitors of the p53-MDM2 interaction could serve as an example for other protein interfaces. PMID- 14757843 TI - MDM2 and its splice variant messenger RNAs: expression in tumors and down regulation using antisense oligonucleotides. AB - Alternative splicing has an important role in expanding protein diversity. An example of a gene with more than one transcript is the MDM2 oncogene. To date, more than 40 different splice variants have been isolated from both tumor and normal tissues. Here, we review what is known about the alteration of MDM2 mRNA expression, focusing on alternative splicing and potential functions of different MDM2 isoforms. We also discuss the progress that has been made in the development of antisense oligonucleotides targeted to MDM2 for use as a potential cancer therapy. PMID- 14757844 TI - Vascular remodeling marks tumors that recur during chronic suppression of angiogenesis. AB - The potential for avoiding acquired resistance to therapy has been proposed as one compelling theoretical advantage of antiangiogenic therapy based on the normal genetic status of the target vasculature. However, previous work has demonstrated that tumors may resume growth after initial inhibition if antiangiogenic blockade is continued for an extended period. The mechanisms of this recurrent growth are unclear. In these studies, we characterized molecular changes in vasculature during apparent resumption of xenograft growth after initial inhibition by vascular endothelial growth factor blockade, "metronome" topotecan chemotherapy, and combined agents in a xenograft murine model of human Wilms' tumor. Tumors that grew during antiangiogenic blockade developed as viable clusters surrounding strikingly remodeled vessels. These vessels displayed significant increases in diameter and active proliferation of vascular mural cells and expressed platelet-derived growth factor-B, a factor that functions to enhance vascular integrity via stromal cell recruitment. In addition, remodeled vessels were marked by expression of ephrinB2, required for proper assembly of stromal cells into vasculature. Thus, enhanced vascular stability appears to characterize tumor vessel response to chronic antiangiogenesis, features that potentially support increased perfusion and recurrent tumor growth. PMID- 14757845 TI - Molecular cloning and characterization of the von Hippel-Lindau-like protein. AB - von Hippel-Lindau (VHL) tumor suppressor protein-inactivated in VHL disease and sporadic kidney cancer-is a component of an E3 ubiquitin ligase complex that selectively ubiquitinates the alpha subunit of the hypoxia-inducible factor (HIF) transcription factor for subsequent destruction by the 26S proteasome. Here, we report the identification and characterization of the first VHL homologue, VHL like protein (VLP), located on chromosome 1q21.2. A 676-bp partial cDNA encoding a 139-amino acid protein that is 78% similar to VHL was isolated by reverse transcription-PCR from human brain cerebellum and several cancer cell lines. The expression of VLP transcript is most abundant in the placenta. Like VHL, VLP contains a beta domain capable of binding HIFalpha. However, unlike VHL, it does not contain a recognizable alpha domain, which is required for nucleating the multiprotein E3 ubiquitin ligase complex. The increased expression of VLP in the presence of VHL attenuated the ubiquitination of HIFalpha and led to the accumulation of downstream HIF target genes. These results taken together indicate that VLP functions as a dominant-negative VHL to serve as a protector of HIFalpha. PMID- 14757846 TI - Topoisomerase I and II inhibitors control caspase-2 pre-messenger RNA splicing in human cells. AB - We have recently shown that the topoisomerase II inhibitor, etoposide (VP16), could trigger caspase-2 pre-mRNA splicing in human leukemic cell lines. This leads to increased inclusion of exon 9, which is specifically inserted into the short caspase-2S isoform mRNA and absent from the long caspase-2L isoform mRNA. One of the consequences of this alternative splicing is a decrease in the total amount of the mature form of caspase-2L mRNA and protein. In this study, we analyzed the effects of several representative molecules of various classes of cytotoxic agents on caspase-2 pre-mRNA splicing in both U937 leukemic cells and in HeLa cervix carcinoma cells. Very strikingly, both topoisomerase I (camptothecin and homocamptothecin derivatives) and II (VP16, amsacrine, doxorubicin, mitoxantrone) inhibitors induced exon 9 inclusion. DNA intercalating glycosyl indolocarbazole derivatives as well as DNA alkylating agents, such as cisplatin and melphalan, antimetabolites like 5-fluorouracil, and mitotic spindle poisons like vinblastine had no effect. Therefore, both classes of DNA topoisomerases can control pre-mRNA splicing of the caspase-2 transcript. In addition, the splicing reaction brought about by camptothecin was hampered in human CEM/C2 and in murine P388-45R leukemic deficient in topoisomerase I activity. Conversely, VP16 did not trigger caspase-2 alternative splicing in human HL60/MX2 leukemic cells harboring a mutant topoisomerase II. Minigene transfection analysis revealed that topoisomerase inhibitors did not change the splicing profile when cis-acting elements in intron-9, reported to control exon 9 inclusion independently of drug treatment, were removed. Rather, our experiments suggest that exon 9 inclusion induced by topoisomerase inhibitors reflects the activity exerted by topoisomerase I or II on proteins that control splicing reactions, or their direct involvement in pre-mRNA splicing. PMID- 14757848 TI - Specific endothelin ET(A) receptor antagonism does not modulate insulin-induced hemodynamic effects in the human kidney, eye, or forearm. AB - There is evidence that hyperinsulinemia may stimulate endothelin-1 (ET-1) generation or release, which may affect diabetic vascular complications. BQ-123, a specific ET(A) receptor antagonist, was used to investigate if insulin-induced vascular effects are influenced by an acute ET-1 release. Two randomized, placebo controlled, double-blind, cross-over studies were performed. In protocol 1, 12 healthy subjects received, on separate study days, infusions of BQ-123 (60 microg/min for 30 min) during placebo clamp conditions, BQ-123 during euglycemic hyperinsulinemia (3 mU/kg/min for 390 min), or placebo during euglycemic hyperinsulinemia. Fundus pulsation amplitude (FPA) was measured to assess pulsatile choroidal blood flow, and mean flow velocity (MFV) of the ophtalmic artery was measured by color Doppler imaging. In protocol 2, eight healthy subjects received, on separate study days, intra-arterial infusions of BQ-123 (32 microg/min for 120 min) during placebo or insulin clamp. Forearm blood flow was measured with bilateral plethysmography, expressing the ratio of responses in the intervention arm and in the control arm. Insulin alone increased FPA (+10%, p < 0.001) and forearm blood flow (+19%). BQ-123 increased FPA, MFV, and forearm blood flow ratio in the absence and presence of exogenous insulin, but this effect was not different between normo- and hyperinsulinemic conditions. ET-1 plasma concentrations were not affected by insulin. In conclusion, these data do not support the concept that hyperinsulinemia increases ET-1 generation in healthy subjects. Our results, however, cannot necessarily be extrapolated to diabetic and obese subjects. PMID- 14757847 TI - Role of the DNA methyltransferase variant DNMT3b3 in DNA methylation. AB - Several alternatively spliced variants of DNA methyltransferase (DNMT) 3b have been described. Here, we identified new murine Dnmt3b mRNA isoforms and found that mouse embryonic stem (ES) cells expressed only Dnmt3b transcripts that contained exons 10 and 11, whereas the Dnmt3b transcripts in somatic cells lacked these exons, suggesting that this region is important for embryonic development. DNMT3b2 and 3b3 were the major isoforms expressed in human cell lines and the mRNA levels of these isoforms closely correlated with their protein levels. Although DNMT3b3 may be catalytically inactive, it still may be biologically important because D4Z4 and satellites 2 and 3 repeat sequences, all known DNMT3b target sequences, were methylated in cells that predominantly expressed DNMT3b3. Treatment of cells with the mechanism-based inhibitor 5-aza-2'-deoxycytidine (5 Aza-CdR) caused a complete depletion of DNMT1, 3a, 3b1, and 3b2 proteins. Human DNMT3b3 and the murine Dnmt3b3-like isoform, Dnmt3b6, were also depleted although less efficiently, suggesting that DNMT3b3 also may be capable of DNA binding. Moreover, de novo methylation of D4Z4 in T24 cancer cells after 5-Aza-CdR treatment only occurred when DNMT3b3 was expressed, reinforcing its role as a contributing factor of DNA methylation. The expression of either DNMT3b2 or 3b3, however, was not sufficient to explain the abnormal methylation of DNMT3b target sequences in human cancers, which may therefore be dependent on factors that affect DNMT3b targeting. Methylation analyses of immunodeficiency, chromosomal instabilities, and facial abnormalities cells revealed that an Alu repeat sequence was highly methylated, suggesting that Alu sequences are not DNMT3b targets. PMID- 14757849 TI - Kappa-opioid receptor ligands inhibit cocaine-induced HIV-1 expression in microglial cells. AB - Cocaine abuse has been implicated as a cofactor in human immunodeficiency virus (HIV)-1-associated dementia (HAD). In this study, we tested the hypothesis that exposure of microglial cells, the resident macrophages of the brain, to cocaine would potentiate HIV-1 expression. Because kappa-opioid receptor (KOR) agonists have been shown to suppress neurochemical and neurobehavioral responses to cocaine and to inhibit HIV-1 expression in microglial cell cultures, we also postulated that KOR ligands would inhibit cocaine-induced potentiation of HIV-1 expression. Human microglial cells were infected with HIV-1(SF162), an R5 isolate, and viral expression was quantified by measurement of p24 antigen in culture supernatants. Treatment of microglia with the KOR agonists trans-(+/-) 3,4-dichlor-N-methyl-N-(2[1-pyrrolidnyl])benzeneacetamide methanesulfonate and 8 carboxamidocyclazocine inhibited viral expression (maximal suppression of 42 and 48%, respectively). Consistent with the hypotheses, treatment of microglia with cocaine promoted HIV-1 expression (maximal enhancement of 54%), and pretreatment of microglia with these KOR agonists as well as with the KOR-selective antagonist nor-binaltorphimine abrogated cocaine-induced potentiation of viral expression. Results of flow cytometry studies suggested that the mechanism whereby KOR ligands inhibit cocaine's stimulatory effect on viral expression involves the suppression of cocaine-induced activation of extracellular signal-regulated kinase1/2, thereby blunting cocaine-enhanced up-regulation of the HIV-1 entry chemokine coreceptor CCR5. The findings of this study suggest that in addition to its neurotoxic effects, cocaine could foster development of HAD by potentiating viral expression in the brain and that this phenomenon is inhibited by KOR ligands. PMID- 14757850 TI - The effect of age on P-glycoprotein expression and function in the Fischer-344 rat. AB - We investigated the effect of age on P-glycoprotein (P-gp) expression and function in rat liver, intestine, kidney, and endothelial cells of the blood brain barrier (BBB) and lymphocytes. Flow cytometric analysis was used to examine P-gp expression in lymphocytes from male Fischer-344 rats from three age groups (young at 3-4 months, intermediate at 13-14 months, and old at 25-26 months). In addition, P-gp function in lymphocytes was assessed by measuring the ability of the P-gp inhibitor verapamil to limit the efflux of the fluorescent P-gp substrate rhodamine 123. P-gp expression was evaluated in the remaining four tissues by Western blot analysis. The effect of age on P-gp expression was tissue specific. Although lymphocytic and hepatic P-gp expression increased with age, renal P-gp content was lower in the old kidneys. No statistical difference was observed in P-gp expression in intestinal microsomes or in BBB cell lysates among the three age groups. P-gp function was also increased by 6- to 8-fold in lymphocytes from the old rats. When P-gp expression was compared with CYP3A expression in these rats (reported elsewhere in this journal), we found that P-gp expression increased with age, whereas CYP3A expression and activity declined in the old livers. The converse pattern was observed in the kidney. Thus, age related changes in P-gp expression and function are likely to be tissue-specific, and these changes may be inversely related to differences in CYP3A expression. PMID- 14757851 TI - Prostaglandin ethanolamides (prostamides): in vitro pharmacology and metabolism. AB - We investigated whether prostaglandin ethanolamides (prostamides) E(2), F(2alpha), and D(2) exert some of their effects by 1) activating prostanoid receptors either per se or after conversion into the corresponding prostaglandins; 2) interacting with proteins for the inactivation of the endocannabinoid N-arachidonoylethanolamide (AEA), for example fatty acid amide hydrolase (FAAH), thereby enhancing AEA endogenous levels; or 3) activating the vanilloid receptor type-1 (TRPV1). Prostamides potently stimulated cat iris contraction with potency approaching that of the corresponding prostaglandins. However, prostamides D(2), E(2), and F(2alpha) exhibited no meaningful interaction with the cat recombinant FP receptor, nor with human recombinant DP, EP(1-4), FP, IP, and TP prostanoid receptors. Prostamide F(2alpha) was also very weak or inactive in a panel of bioassays specific for the various prostanoid receptors. None of the prostamides inhibited AEA enzymatic hydrolysis by FAAH in cell homogenates, or AEA cellular uptake in intact cells. Furthermore, less than 3% of the compounds were hydrolyzed to the corresponding prostaglandins when incubated for 4 h with homogenates of rat brain, lung, or liver, and cat iris or ciliary body. Very little temperature-dependent uptake of prostamides was observed after incubation with rat brain synaptosomes or RBL-2H3 cells. We suggest that prostamides' most prominent pharmacological actions are not due to transformation into prostaglandins, activation of prostanoid receptors, enhancement of AEA levels, or gating of TRPV1 receptors, but possibly to interaction with novel receptors that seem to be functional in the cat iris. PMID- 14757852 TI - Phytoestrogen cimicifugoside-mediated inhibition of catecholamine secretion by blocking nicotinic acetylcholine receptor in bovine adrenal chromaffin cells. AB - We investigated the effect of the phytoestrogen cimicifugoside, one of the pharmacologically active ingredients of the medicinal plant Cimicifuga racemosa (black cohosh) that has been used to treat many kinds of neuronal and menopausal symptoms, such as arthritis, menopausal depression, and nerve pain. Cimicifugoside inhibited calcium increase induced by 1,1-dimethyl-4 phenylpiperazinium iodide (DMPP), a nicotinic acetylcholine receptor (nAChR) agonist in bovine adrenal chromaffin cells with a half-maximal inhibitory concentration (IC(50)) of 18 +/- 2 microM. In contrast, cimicifugoside did not affect the calcium increases evoked by high K(+), veratridine, and bradykinin. The DMPP-induced sodium increase was also inhibited by cimicifugoside with an IC(50) of 2 +/- 0.3 microM, suggesting that the activity of nAChRs is inhibited by cimicifugoside. Cimicifugoside did not affect the KCl-induced secretion but markedly inhibited the DMPP-induced catecholamine secretion that was monitored by carbon-fiber amperometry in real time and high-performance liquid chromatography through electrochemical detection. The results suggest that cimicifugoside selectively inhibits nAChR-mediated response in bovine chromaffin cells. PMID- 14757853 TI - Referral for cancer genetics consultation: a review and compilation of risk assessment criteria. AB - BACKGROUND: There have been many papers on the diagnostic criteria for specific hereditary cancer susceptibility syndromes and the likelihood that an individual has a germline mutation in one of the various cancer susceptibility genes. To assist health care professionals in deciding when a cancer genetics consultation is appropriate, available reports were critically reviewed in order to develop a single set of risk assessment criteria. METHODS: The criteria were based on a comprehensive review of publications describing diagnostic criteria for hereditary cancer syndromes and risk to first degree relatives of cancer patients. Priority was given to diagnostic criteria from consensus statements (for example, those from the National Comprehensive Cancer Network). Expert opinion from study personnel was then used to adopt a single set of criteria from other publications whenever guidelines differed. RESULTS: Based on family history, a set of criteria was developed to identify patients at risk for a hereditary cancer susceptibility syndrome, patients with moderate risk who might benefit from increased cancer surveillance, and patients who are at average risk. The criteria were applied to 4360 individuals who provided their cancer family history between July 1999 and April 2002, using a touch screen computer system in the lobby of a comprehensive cancer centre. They categorised an acceptable number of users into each risk level: 14.9% high risk, 13.7% moderate risk, and 59.6% average risk; 11.8% provided insufficient information for risk assessment. CONCLUSIONS: These criteria should improve ease of referral and promote consistency across centres when evaluating patients for referral to cancer genetics specialists. PMID- 14757854 TI - Mulibrey nanism: clinical features and diagnostic criteria. AB - Mulibrey nanism (MUL) is an autosomal recessive disease caused by mutations in the TRIM37 gene encoding the peroxisomal TRIM37 protein of unknown function. In this work, we analysed the clinical characteristics of 85 Finnish patients with MUL, most of whom were homozygous for the Finn major mutation of TRIM37. The patients' hospital records from birth to the time of the diagnosis at age 0.02-52 years (median 2.1 years) were retrospectively analysed. All except four of the patients (95%) had a prenatal onset growth failure without postnatal catch up growth. The mean length standard deviation score (SDS) was -3.1 and -4.0 at birth and at diagnosis, respectively. In infancy, feeding difficulties, and respiratory tract infections were the most common problems. Congestive heart failure and pericardial constriction were diagnosed during infancy in 12% and 6% of the patients, respectively. At the time of the diagnosis, characteristic craniofacial features of scaphocephaly, facial triangularity, high and broad forehead, and low nasal bridge were evident in over 90% of the patients. In addition, practically all patients were gracile and had thin extremities. Other findings included a peculiar high-pitched voice (96%), yellowish dots in ocular fundi (79%), cutaneous naevi flammei (65%), hepatomegaly (45%), and fibrous dysplasia of long bones (25%). Mild muscular hypotonicity (68%) was the only neurological abnormality. The clinical features of the Finnish patients with MUL formed a distinct entity. The most consistent findings were growth failure and characteristic craniofacial features. However, organ manifestations varied considerably in early childhood. Based on these findings, we propose new diagnostic criteria for MUL. PMID- 14757856 TI - A study of gene--environment interaction on the gene for angiotensin converting enzyme: a combined functional and population based approach. AB - INTRODUCTION: Studies on the role of the insertion/deletion (I/D) polymorphism of the gene coding for angiotensin converting enzyme (ACE) in atherosclerosis have been inconsistent. In a meta-analysis, we recently showed that this relationship is stronger in high risk populations. In this paper, we used a combined functional and population based approach to investigate the gene-environment interaction of the ACE I/D polymorphism in relation to carotid artery wall thickness. METHODS: The study was part of the Rotterdam Study, a prospective population based cohort study. In 5321 subjects, IMT was measured in the carotid arteries by ultrasonography and ACE genotype was determined by size analysis of polymerase chain reaction products. RESULTS: In multiple regression analysis, I/D polymorphism and smoking were the main determinants for plasma ACE activity (r(2) = 0.28). There was a positive association between the D allele of the I/D polymorphism and carotid artery thickness among current smokers (p = 0.03). Subjects carrying only one of the risk factors (smoking or the D allele) did not show significant differences in IMT compared with the non-/former smokers group carrying two II alleles, while carriers of both risk factors had significant higher IMT. The association was not present in non-/former smokers. DISCUSSION: The results provide further evidence that genetic and environmental factors interact in the formation of the arterial lesions. This study shows that large population based studies can be extremely helpful in unravelling the genetic origin of complex diseases such as atherosclerosis. PMID- 14757858 TI - Comparative genomic hybridisation using a proximal 17p BAC/PAC array detects rearrangements responsible for four genomic disorders. AB - BACKGROUND: Proximal chromosome 17p is a region rich in low copy repeats (LCRs) and prone to chromosomal rearrangements. Four genomic disorders map within the interval 17p11-p12: Charcot-Marie-Tooth disease type 1A, hereditary neuropathy with liability to pressure palsies, Smith-Magenis syndrome, and dup(17)(p11.2p11.2) syndrome. While 80-90% or more of the rearrangements resulting in each disorder are recurrent, several non-recurrent deletions or duplications of varying sizes within proximal 17p also have been characterised using fluorescence in situ hybridisation (FISH). METHODS: A BAC/PAC array based comparative genomic hybridisation (array-CGH) method was tested for its ability to detect these genomic dosage differences and map breakpoints in 25 patients with recurrent and non-recurrent rearrangements. RESULTS: Array-CGH detected the dosage imbalances resulting from either deletion or duplication in all the samples examined. The array-CGH approach, in combination with a dependent statistical inference method, mapped 45/46 (97.8%) of the analysed breakpoints to within one overlapping BAC/PAC clone, compared with determinations done independently by FISH. Several clones within the array that contained large LCRs did not have an adverse effect on the interpretation of the array-CGH data. CONCLUSIONS: Array-CGH is an accurate and sensitive method for detecting genomic dosage differences and identifying rearrangement breakpoints, even in LCR-rich regions of the genome. PMID- 14757859 TI - Respiratory chain complex V deficiency due to a mutation in the assembly gene ATP12. AB - In patients with mitochondrial encephalomyopathies an increasing number of causative gene defects have been detected. The number of identified pathogenic mitochondrial DNA mutations has largely increased over the past 15 years. Recently, much attention has turned to the investigation of nuclear oxidative phosphorylation (OXPHOS) gene defects. Within the OXPHOS defects, complex V deficiency is rarely found and, so far, these defects have only been attributed to mutations in the mitochondrial MTATP6 gene. Mutation analysis of the complete coding regions at the cDNA level of the nuclear ATP11, ATP12, ATPalpha, ATPbeta and ATPgamma genes and the mitochondrial MTATP6 and MTAT8 genes was undertaken in two unrelated patients. Blue Native polyacrylamide gel electrophoresis followed by catalytic staining had already documented their complex V decreased activity. Extensive molecular analysis of five nuclear and two mitochondrial genes revealed a mutation in the ATP12 assembly gene in one patient. This mutation is believed to be the cause of the impaired complex V activity. To our knowledge, this is the first report of a pathogenic mutation in a human nuclear encoded ATPase assembly gene. PMID- 14757860 TI - MNGIE with lack of skeletal muscle involvement and a novel TP splice site mutation. AB - Mitochondrial neurogastrointestinal encephalomyopathy (MNGIE) is an autosomal recessive multisystem disorder caused by thymidine phosphorylase (TP) deficiency, resulting in severe gastrointestinal dysmotility and skeletal muscle abnormalities. A patient is reported with a classical MNGIE clinical presentation but without skeletal muscle involvement at morphological, enzymatic, or mitochondrial DNA level, though gastrointestinal myopathy was present. MNGIE was diagnosed by markedly raised plasma thymidine and reduced thymidine phosphorylase activity. Molecular genetic analysis showed a homozygous novel splice site mutation in TP. On immunohistochemical studies there was marked TP expression in the CNS, in contrast to what has been observed in rodents. It is important to examine the most significantly affected tissue and to measure TP activity and plasma thymidine in order to arrive at an accurate diagnosis in this condition. PMID- 14757857 TI - The allelic modulation of apolipoprotein E expression by oestrogen: potential relevance for Alzheimer's disease. AB - BACKGROUND: The epsilon4 allele of the apolipoprotein E (APOE) gene is a major genetic risk factor for Alzheimer's disease but appears to be associated with greater risk in women than in men. Some studies suggest that the level of APOE may of its own modulate the risk for Alzheimer's disease. Sex differences and an apparent benefit of oestrogen therapy suggest a role for oestrogen. APOE expression is influenced by oestrogen and oestrogen therapy may not benefit women bearing an APOE epsilon4 allele. These findings suggest an interaction between oestrogen and APOE in the Alzheimer's disease process. AIM: To explore the hypothesis that APOE expression is regulated by a genomic mechanism and is modified by the polymorphisms in APOE associated with risk for Alzheimer's disease. METHODS: In vitro binding studies were undertaken between oestrogen receptors and fragments of the human APOE gene. APOE gene expression was studied to investigate a possible functional interaction. RESULTS: APOE epsilon2/epsilon3/epsilon4 coding and -219 G/T promoter polymorphisms influenced binding to the oestrogen receptor and altered transcriptional activity in response to oestrogen. CONCLUSIONS: An allele dependent modulation of oestrogen induced regulation of APOE might be involved in the increased risk for Alzheimer's disease in women bearing an epsilon4 allele. PMID- 14757861 TI - Subtelomere specific microarray based comparative genomic hybridisation: a rapid detection system for cryptic rearrangements in idiopathic mental retardation. PMID- 14757862 TI - Characterisation of diverse PRF1 mutations leading to decreased natural killer cell activity in North American families with haemophagocytic lymphohistiocytosis. PMID- 14757863 TI - Melanocortin 1 receptor (MC1R) gene variants may increase the risk of melanoma in France independently of clinical risk factors and UV exposure. PMID- 14757864 TI - A novel locus for autosomal dominant nonsyndromic hearing loss, DFNA50, maps to chromosome 7q32 between the DFNB17 and DFNB13 deafness loci. PMID- 14757865 TI - Six families with van der Woude and/or popliteal pterygium syndrome: all with a mutation in the IRF6 gene. PMID- 14757866 TI - Mowat-Wilson syndrome and mutation in the zinc finger homeo box 1B gene: a well defined clinical entity. PMID- 14757867 TI - Homozygosity for autosomal dominant facioscapulohumeral muscular dystrophy (FSHD) does not result in a more severe phenotype. PMID- 14757868 TI - Loss of five amino acids in BRCA2 is associated with ovarian cancer. PMID- 14757869 TI - Mitochondrial DNA deletion in "identical" twin brothers. PMID- 14757871 TI - Haplotype and cancer risk analysis of two common mutations, BRCA1 4184del4 and BRCA2 2157delG, in high risk northwest England breast/ovarian families. PMID- 14757870 TI - Achromatopsia caused by novel mutations in both CNGA3 and CNGB3. PMID- 14757872 TI - Advances in stroke 2003: introduction. PMID- 14757873 TI - Genetics of cerebrovascular disease. PMID- 14757874 TI - Hyperhomocysteinemia, oxidative stress, and cerebral vascular dysfunction. PMID- 14757875 TI - Hypertension, angiotensin, and stroke: beyond blood pressure. PMID- 14757876 TI - Neuroimaging. PMID- 14757877 TI - tPA and proteolysis in the neurovascular unit. PMID- 14757878 TI - Reborn workhorse, CT, pulls the wagon toward thrombolysis beyond 3 hours. PMID- 14757879 TI - Acute neurovascular syndromes: hurry up, please, it's time. PMID- 14757880 TI - Hyperglycemia in acute stroke. PMID- 14757881 TI - Critical care and emergency medicine neurology. PMID- 14757882 TI - Emerging therapies for cerebrovascular disorders. PMID- 14757883 TI - Is ultrasound sufficient for vascular imaging prior to carotid endarterectomy? PMID- 14757884 TI - Treatment options of unruptured intracranial aneurysms. PMID- 14757885 TI - Advances in vascular surgery. PMID- 14757886 TI - Interventional neuroradiology. PMID- 14757887 TI - What's new in stroke rehabilitation. PMID- 14757888 TI - Vascular cognitive impairment. PMID- 14757889 TI - Ximelagatran or warfarin for stroke prevention in patients with atrial fibrillation? PMID- 14757890 TI - Evidence-based clinical practice education in cerebrovascular disease. PMID- 14757891 TI - Health policy and outcome research in stroke. PMID- 14757892 TI - Prevention and health services delivery. PMID- 14757893 TI - Stroke in a biracial population: the excess burden of stroke among blacks. AB - BACKGROUND AND PURPOSE: Excess mortality resulting from stroke is an important reason why blacks have higher age-adjusted mortality rates than whites. This observation has 2 possible explanations: Strokes occur more commonly among blacks or blacks have higher mortality rates after stroke. Our population-based epidemiological study is set in the Greater Cincinnati/Northern Kentucky region of 1.31 million people, which is representative of the US white and black populations with regard to many demographic and socioeconomic characteristics. METHODS: Hospitalized cases were ascertained by International Classification of Diseases (ninth revision) discharge codes, prospective screening of emergency department admission logs, and review of coroner's cases. A sampling scheme was used to ascertain cases in the out-of-hospital setting. All potential cases underwent detailed chart abstraction by study nurses, followed by physician review. Race-specific incidence and case fatality rates were calculated. RESULTS: We identified 3136 strokes during the study period (January 1, 1993, to June 30, 1994). Stroke incidence rates were higher for blacks at every age, with the greatest risk (2- to 5-fold) seen in young and middle-aged blacks (<65 years of age). Case fatality rates did not differ significantly in blacks compared with whites. Applying the resulting age- and race-specific rates to the US population in 2002, we estimate that 705,000 to 740,000 strokes have occurred in the United States, with a minimum of 616,000 cerebral infarctions, 67,000 intracerebral hemorrhages, and 22,000 subarachnoid hemorrhages. CONCLUSIONS: Excess stroke related mortality in blacks is due to higher stroke incidence rates, particularly in the young and middle-aged. This excess burden of stroke incidence among blacks represents one of the most serious public health problems facing the United States. PMID- 14757894 TI - Mechanism of ischemic infarct in spontaneous carotid dissection. AB - BACKGROUND AND PURPOSE: It is unclear whether stroke in patients with spontaneous dissection of the cervical internal carotid artery (ICAD) is due to thromboembolism or impaired hemodynamics. This study investigated the mechanism of stroke in ICAD by examining brain imaging and cerebrovascular findings of such patients. METHODS: We retrospectively evaluated the prospectively collected brain CT, MR, and ultrasound findings of 141 consecutive patients with 143 ICADs causing ischemic stroke. Eleven patients were not included because they had an inappropriate temporal bone window (n=6) or were treated with thrombolysis (n=5). Thus, the data of 130 patients (76 men, 54 women) with 131 ICADs were analyzed. RESULTS: All patients had territorial infarcts; 6 patients (5%) also had border zone infarct patterns. Territorial infarcts affected the middle cerebral artery (MCA) in 130 of 131 cases (99%) and the anterior cerebral artery (ACA) in 1 case (1%). Additional vascular territories were affected in 8 patients with MCA infarcts (ACA, n=5 [4%]; posterior cerebral artery, n=3 [2%]). The pattern (hemodynamic versus thromboembolic) and extent of infarction were not influenced by vascular findings (MCA stenosis or occlusion, ACA occlusion, degree of obstruction in the dissected ICA, pattern of cross-flow in 115 patients with >80% ICA stenosis or occlusion). CONCLUSIONS: This study suggests that thromboembolism, not hemodynamic infarction, is the essential stroke mechanism in ICAD. PMID- 14757895 TI - Editorial comment--an approach to the estimation of the risk of TTP during clopidogrel therapy. PMID- 14757896 TI - Editorial comment--myocardial damage in patients with subarachnoid hemorrhage. PMID- 14757897 TI - Major ongoing stroke trials. PMID- 14757899 TI - A new approach in vocational rehabilitation in Iceland: preliminary report. AB - The lack of vocational rehabilitation in Iceland inspired the Janus Rehabilitation initiative in the year 2000. The team currently consists of two occupational therapists, a social worker, a physiotherapist, a psychologist, and a physician. Janus uses resources from local education establishments and is located at the Reykjavik Technical School. A client-centred approach is used, enabling a flexible rehabilitation timeframe. The aim is to help the client back to employment/education. Advantage is taken of the facilities of the school where integration between education and rehabilitation takes place. Forty individuals have been admitted, with a mean time from work of 2.9 years, (range: 0-11 years). Seventeen (43%) have returned to employment/education. Twenty-three of those entering the programme are still on invalidity pension. The Canadian Occupational Performance Measurement (COPM) shows improvement in occupational performance and satisfaction. The Icelandic Quality of Life measurement also shows improvement. The results have been promising. However, a larger group is needed in order to draw clear unequivocal conclusions. PMID- 14757900 TI - Outcome of an interdisciplinary pain management program in a rehabilitation clinic. AB - The Chronic Pain Section at Reykjalundur has 33 beds for patients with various chronic pain problems. About 200 patients are treated annually. For 3 years, 158 patients were enrolled in a random study focusing on increasing the patients' functioning and eliminating analgesic drugs through an interdisciplinary rehabilitation program. Patients answered a questionnaire at admission, before discharge from the clinic and about one year after discharge. The follow-up was done with a mailed questionnaire. The focus was on the patients' functioning rather than absence of pain. The program's duration was 7 weeks. The first 2 weeks were utilized for evaluation, information and education (pain school). Pain relieving drugs were gradually withdrawn but anti-inflammatory drugs were used when indicated. Cognitive behavioral therapy was applied in an increasing number of cases over the 3 year period by specially trained nurses and a psychiatrist. Pain, anxiety and depression were self-evaluated on a numeric rating scale (NRS). About 50% of the patients had a history of pain for more than 5 years. Low back pain was the most common diagnosis (48.1%) and 28.5% had post-traumatic pain. A significant reduction in pain, anxiety and depression was found both at discharge (p < 0.0001) and at follow-up (p < 0.001). Before entering the program, only 18.4% of the patients were able to work whereas 48.1% returned to work after discharge and 59.2% were working at follow-up. PMID- 14757902 TI - The Worker Role Interview: a powerful tool in Icelandic work rehabilitation. AB - A pilot-study using the Worker Role Interview (WRI) was conducted to gather information from clients attending a work- rehabilitation program in an Icelandic psychiatric center. The work rehabilitation program traditionally used a performance-capacity-oriented assessment (PCA) to evaluate work readiness. Clients scoring 90% or higher on the PCA were considered able to re-enter the job market. However, scoring over 90% did not result in automatic return to work for the majority of clients. As a result, many questions were proposed about the effectiveness of the PCA to predict client ability to re-enter the work force. A pilot study using the Worker Role Interview was conducted with an aim to gain insight into what barriers prevented clients from entering the work force. Ten clients scoring 90% or over on the PCA were interviewed using the WRI. Qualitative methods were used to analyze the data. Results from the WRI indicated that clients failed to join the work force due to a number of psychosocial factors, such as lack of personal causation and self-efficacy combined with environmental factors. The WRI provided clients with an opportunity to express their attitudes and opinions towards work. Information gained through the WRI also gave new direction in goal setting for the work rehabilitation program and validated the need to expand occupational therapy services. PMID- 14757901 TI - Vocational rehabilitation at Reykjalundur Rehabilitation Center in Iceland. AB - This article describes a pilot project in specialized vocational rehabilitation program at Reykjalundur Rehabilitation Center in Iceland. It gives a brief historical overview of the development of the program, describes the theoretical frame used and the way the program is designed. Finally, it discusses the results of the program and ways to improve the program. PMID- 14757903 TI - Prevention, health and safety program in companies provide a more successful and healthier workplace. AB - This article describes the importance of injury prevention in the workplace and the status of this issue in Iceland. Using a theoretical approach, this article examines workers' psychological and physical health, as it is affect by work. The article seeks to answer the question, "What methods are appropriate when health and safety management is of primary importance in the workplace?" Finally, the article will explain the condition of these matters in Iceland, and will also discuss how one measurement strategy, called "Workplace Analysis", uses performance indicators to demonstrate the effectiveness of health and safety consultations. This tool has been developed by Solarpexus. It is important to analyze the work environment, set goals, and measure results. PMID- 14757904 TI - New technology and its impact on well being. AB - Great changes have come about in the last decade regarding the organization of work. High technology and the steadily increasing ideology of technocracy has produced a profound effect on the organization of work in some workplaces. This effect has made the psychosocial and physical working environment tougher, especially for women. The results we are presenting here show that when dividing fishing factories into three technological stages; low technology, middle technology and high technology, the job strain was highest and the decision authority by the employee was lowest in the high technological factories. This even had an impact on health and on the atmosphere at the workplace, where the employees in the high technological factories were more likely to complain about several health problems as well as about low degrees of cheerfulness at the workplace and tiresome jobs. However, these same people were the most positive towards the implementation of the high technology and the new way of organizing their job that the technology introduced. PMID- 14757905 TI - Well-being and self-assessed health among different groups of female personnel in geriatric care. AB - Educational qualifications are reliable predictors of women's self-assessed health. AIMS: To study possible inequalities in health among women with different educational backgrounds working in geriatric care and to find groups that might need special public health measures. METHODS: In this cross-sectional questionnaire reaching throughout Iceland, the participants were employees in 62 geriatric nursing homes and geriatric hospital wards with 10 or more employees. A total of 1886 questionnaires were distributed. The 84-item questionnaire included questions on demographic and work-related factors, health and life style. Age adjusted odds ratios (OR) were calculated for work-related psychosocial, physical and health factors, and confidence intervals were set at 95% (95% CI). Registered nurses were taken as a reference category. RESULTS: The response rate was 80%. Registered nurses accounted for 16%, practical nurses 21%, unskilled attendants 44%, cleaning personnel 8% and others 12%. The practical nurses, unskilled attendants and cleaning personnel assessed work as more physically difficult, and more monotonous both physically and mentally, than did the registered nurses, who enjoyed more physical and mental well-being than the others. However, the registered nurses visited doctors as often as the other groups did. CONCLUSIONS: Personnel groups in geriatric care have different physical and psychosocial workloads. The results provide opportunities to guide public health measures for people employed in geriatric care and possibly in other settings, such as hospitals and health care institutions. PMID- 14757906 TI - Work organization, well-being and health in geriatric care. AB - The objective of the present study was to explore the connection between the organization of work in geriatric care and factors which have been connected to job stress and burnout, i.e. exhaustion, mental workload, job satisfaction and communication. We also analyzed how these factors were related to employee visits to doctors during the previous 12 months due to various medical conditions. The study was a cross-sectional questionnaire distributed to all employees within nursing homes and geriatric hospital wards with 10 employees or more throughout Iceland. The total response rate was 80%. The majority of respondents, or 96%, were women (n = 1432), and the results are based on their answers. Our data show that there is a high correlation between mental exhaustion and the unsatisfactory organization of work. Mental exhaustion upon completing work shifts was more closely connected to the health outcomes studied than were the other work-related factors studied. This is especially true for chronic fatigue, depression and sleeping disorders. It is important that employers and managers notice the mismatches between work and workers that this study manifests. Employers and managers must also consider the organizational factors that are influential. PMID- 14757907 TI - The relationship between fishermen's health and sleeping habits. AB - This article describes research on fishermen's health and sleeping habits in Iceland. It presents the first findings of the project that were presented in October 2002 at a conference that was organized during Seamen Safety Week in Iceland. It covers physical and mental stress symptoms, sleep disturbance, and obesity, as well as fishermen's own reaction to their health and well-being. Take notice that the findings that are presented in this article can be found in a report presented in August, when the whole study will be finished. Finally, it is concluded how successful the response has been and how the project can be used positively with other studies. It is also noted that, in fact, these findings will be used for further studies linked to safety on board. PMID- 14757908 TI - Mortality among female industrial workers in Iceland. AB - AIMS: The aim was to describe the mortality pattern among unskilled female industrial workers with the hypothesis that they were disadvantaged compared to other women and that smoking-related causes of death were in excess among them. METHODS: The study group comprised 13349 women who had contributed at any time to a pension fund for unskilled industrial workers in Reykjavik during the time period 1970-1995. The follow-up was from 1975 to 1995. The death rate of the study group was compared to that of women in the general population during the time period 1981-1995. The study group was studied with regard to selected causes of death, by age at first entrance into the pension fund, different lag-time, and by employment-time. RESULTS: Results showed an excess of external causes but a deficit of most other causes of death, among those smoking related diseases. The high mortality by external causes was consistent in all the analyses. Standardised mortality ratio (SMR) for external causes in the total group was 1.79 (95% CI 1.45-2.19), for lung cancer 1.04 (95% CI 0.80-1.34) and for ischemic heart disease 0.77 (95% CI 0.65-0.91). Mortality was highest among those who started to pay in the pension fund at younger ages. CONCLUSIONS: The notable excess of external causes in the group needs further exploration. The results did not confirm the hypothesis that smoking-related causes of death were in excess in the group. Methodological problems related to studies on women workers are discussed. PMID- 14757909 TI - Respiratory control during postural changes in anesthetized cats. AB - The effect of postural changes on respiration was investigated in ten anesthetized cats by applying body tilting during the inspiration phase while recording respiratory patterns, as given by the diaphragmatic EMG, together with either the lung volume or the air flow temperature. The results show that the head-up tilting during inspiration reduced the period of the inspiratory phase and increased the end-inspiratory lung volume. On the other hand, the head-down tilting during inspiration had opposite effects. These effects disappeared after transection of the vagus nerve. However, labyrinthectomy did not diminish the effects, probably because of functional suppression of the vestibular system due to the anesthetic. When correlating the activity of 15 vagal afferents presumably originating from the slowly adapting lung stretch receptors with lung volume changes during tilting, their maximum firing rate (87 +/- 15.7 Hz) was increased with an increase in the lung inflation volume and was attained earlier on head-up tilting and it was reduced with a decrease of the lung volume on head-down tilting (63 +/- 16.6 Hz) as compared with the value in the horizontal position (74 +/- 14.2 Hz). These results suggest that respiratory modulation during head up or head-down tilting is consistent with the Hering-Breuer reflexes and minimizes the externally induced lung volume changes during postural changes. PMID- 14757910 TI - Adaptation of the vestibulo-ocular reflex, subjective tilt, and motion sickness to head movements during short-radius centrifugation. AB - Head movements made while the whole body is rotating at unusually high angular velocities (here with supine body position about an earth-vertical axis) result in inappropriate eye movements, sensory illusions, disorientation, and frequently motion sickness. We investigated the acquisition and retention of sensory adaptation to cross-coupled components of the vestibulo-ocular reflex (VOR) by asking eight subjects to make headturns while being rotated at 23 rpm on two consecutive days, and again a week later. The dependent measures were inappropriate vertical VOR, subjective tilt, and motion sickness in response to 90 degrees yaw out-of-plane head movements. Motion sickness was evaluated during and following exposure to rotation. Significant adaptation effects were found for the slow phase velocity of vertical nystagmus, the reported magnitude of the subjective tilt experienced during head turns, and motion-sickness scores. Retention of adaptation over a six-day rest period without rotation occurred, but was not complete for all measures. Adaptation of VOR was fully maintained while subjective tilt was only partially maintained and motion-sickness scores continued to decrease. Practical implications of these findings are discussed with particular emphasis on artificial gravity, which could be produced in weightlessness by means of a short-radius (2 m) rotator. PMID- 14757911 TI - Eye-head coordination in darkness: formulation and testing of a mathematical model. AB - Passive head rotation in darkness produces vestibular nystagmus, consisting of slow and quick phases. The vestibulo-ocular reflex produces the slow phases, in the compensatory direction, while the fast phases, in the same direction as head rotation, are of saccadic origin. We have investigated how the saccadic components of the ocular motor responses evoked by active head rotation in darkness are generated, assuming the only available sensory information is that provided by the vestibular system. We recorded the eye and head movements of nine normal subjects during active head rotation in darkness. Subjects were instructed to rotate their heads in a sinusoidal-like manner and to focus their attention on producing a smooth head rotation. We found that the desired eye position signal provided to the saccadic mechanism by the vestibular system may be modeled as a linear combination of head velocity and head displacement information. Here we present a mathematical model for the generation of both the slow and quick phases of vestibular nystagmus based on our findings. Simulations of this model accurately fit experimental data recorded from subjects. PMID- 14757912 TI - Neurovestibular symptoms following space flight. AB - Neurovestibular symptoms experienced by astronauts in the post-flight period were examined using data from medical debriefs contained in the NASA Longitudinal Study of Astronaut Health database. Ten symptoms were identified (clumsiness, difficulty concentrating, persisting sensation aftereffects, nausea, vomiting, vertigo while walking, vertigo while standing, difficulty walking a straight line, blurred vision, and dry heaves), of which eight were crossed with twelve demographic parameters (mission duration, astronaut gender, age, one-g piloting experience, previous space flight experience, g-suit inflation, g-suit deflation, in-flight space motion sickness, in-flight exercise, post-flight exercise, mission role, fluid loading). Three symptoms were experienced by a majority of subjects, and another two by more than a quarter of the subjects. Intensity of the symptoms was mild, suggesting that they are unlikely to pose a risk to the crew during landing and the post-flight period. Seven of the symptoms and eight of the parameters under study were found to be significantly associated with each other. PMID- 14757913 TI - Evaluation of betahistine for the prevention of seasickness: effect on vestibular function, psychomotor performance and efficacy at sea. AB - Betahistine was evaluated for the prevention of seasickness in a laboratory and sea study. The effect of 48 mg betahistine on the vestibulo-ocular reflex (VOR) and on psychomotor performance was evaluated in twelve young healthy subjects in a double-blind, placebo controlled, randomized, crossover design. The vestibulo ocular reflex was evaluated by the Sinusoidal Harmonic Acceleration (SHA) test at frequencies of 0.01, 0.02, 0.04, 0.08 and 0.16 Hz. Psychomotor performance was assessed by both computerized and paper and pencil test batteries. No significant differences in VOR gain or phase were found between betahistine and placebo treatment for any of the frequencies tested. No significant differences were found between treatments for any of the psychomotor performance tests or other possible side effects. The effect of 48 mg betahistine on seasickness severity was evaluated in 83 subjects during a voyage in rough seas. Betahistine had a borderline non-statistically significant effect on the prevention of seasickness in comparison with placebo (p = 0.053), with no notable side effects. Although our results are insufficient to recommend betahistine as an anti-seasickness drug, further studies are required to determine its possible effectiveness in less provocative motion sickness situations. PMID- 14757914 TI - A comparison of the monothermal and bithermal caloric tests. AB - This study considered whether the monothermal (MT) caloric test could predict the normality of the full conventional bithermal (BT) caloric test, and therefore be an alternative to full caloric investigation. This would have the advantages of reducing test time and patient discomfort as only two caloric tests would be needed instead of four. 744 BT caloric investigations were examined, and the unilateral weakness and directional preponderance calculated for the BT and the MT stimuli. By defining the BT results as the standard, the false-positive and false-negative results of the MT test were derived. Overall using very strict MT difference criteria of less than 5% and no spontaneous nystagmus, false-negative rates for the cool MT were very low (< 1%) and better than the warm MT (< 7.1%) suggesting that the cool MT was a reliable screen test. However, unacceptably high false-positive rates were produced reflecting more than 3/4 of normal BT results failing the MT criterion. This unacceptable false-positive rate decided against implementing the MT test at our facility. The results of this study however have guided the use of the cool air-stimulus first during BT testing and, when completion of the BT is not possible or inadvisable, satisfying the stringent MT criterion confidently indicates with a probability of > 99% the absence of an abnormal BT result. PMID- 14757915 TI - Vestibular evoked myogenic potentials in response to skull taps for patients with vestibular neuritis. AB - In recent years it has been demonstrated that loud clicks generate short latency vestibular evoked myogenic potentials (VEMP). It has also been demonstrated that skull tap stimulation evokes similar VEMP. In the present study, the differences between the click-induced and the skull-tap induced VEMP were studied in 18 patients at onset of vestibular neuritis. Gentle skull taps were delivered manually above each ear on the side of the skull and on the forehead midline. The muscular responses were recorded over both sternocleidomastoid muscles using skin electrodes. Abnormal skull tap VEMP were found in the majority of the patients (10/18, 56%). However, only 4/18 (22%) showed asymmetry in the click-induced VEMP. The high percentage of abnormal skull tap VEMP might suggest that this response is not only dependent on the inferior division of the vestibular nerve, because the inferior division of this nerve is usually spared in vestibular neuritis. Moreover, the patients with abnormal skull tap VEMP differed from those with normal VEMP in their settings of the subjective visual horizontal with static head tilt in the roll plane. This might suggest that skull tap VEMP are (also) related to utricular function. PMID- 14757916 TI - Eye movement responses to active, high-frequency pitch and yaw head rotations in subjects with unilateral vestibular loss or posterior semicircular canal occlusion. AB - This study assessed the eye movement responses to active head rotation in six subjects with complete unilateral vestibular loss (UVL), five subjects with posterior canal plugging (PCP) and age- and sex-matched normal subjects. Subjects performed head rotations in the pitch and yaw planes at frequencies ranging from 2 to 6 Hz, while looking at an earth-fixed target. Vertical eye movement gains obtained in UVL, PCP and normal subjects were not significantly different. Vertical phases decreased with increasing head movement frequencies in both UVL and PCP subjects. Although this decrease produced significantly different vertical phases between UVL and normal subjects for head movements above 3.9 Hz, vertical phases in some normal subjects were similar to those obtained in UVL subjects. We conclude that active head oscillations in the pitch plane are not clinically useful for the detection of vertical canal impairment limited to one ear. As expected, UVL subjects showed reduced horizontal gains, and eye velocity asymmetries during active head rotation in the yaw plane. Results in some PCP subjects suggested possible minor impairments of horizontal vestibulo-ocular reflexes. PMID- 14757917 TI - Otolith-ocular responses in patients with surgically confirmed unilateral peripheral vestibular loss. AB - The chronic effects of unilateral peripheral vestibular loss (UPVL) are influenced by vestibular compensation. This study documents the balance-related symptoms and quantitative vestibular laboratory testing of 20 patients with surgically confirmed UPVL. Included are measures of the semicircular canal-ocular reflex, the otolith-ocular reflex, and both static and dynamic semicircular canal otolith-interaction. This study differs from previous studies of patients with UPVL in that a large number of patients with surgically confirmed lesions were tested with three types of off-vertical axis rotation, several of the patients had anatomically preserved superior vestibular nerves, and self-perceived level of disability related to dizziness and imbalance were available. Results confirmed previously reported changes in the vestibulo-ocular reflex of patients with UPVL. Also, there was no apparent effect of anatomically preserving the superior vestibular nerve during surgical resection of vestibular schwannomas based on either subjective or objective measures of vestibular dysfunction. Further, there were no apparent correlations between subjective measures of dizziness and imbalance and objective measures of vestibulo-ocular function. These results have clinical implications for the management of patients with unilateral vestibular loss and provide insights into the process of vestibular compensation especially with respect to the otolith-ocular reflex. PMID- 14757918 TI - Minimum technical requirements and evaluating effectiveness of teleradiology. AB - Teleradiology, the electronic transmission of radiological images for the purposes of interpretation and consultation, has successfully entered routine health services. It is, therefore, currently one of the single largest applications of telemedicine. This technology is predicted to play a significant role in improving service access to rural and remote areas. However, teleradiology services are currently characterized by a compromise between expense and quality of digital images. This paper reviews minimum equipment requirements and the technical issues and limits that are experienced when utilizing current teleradiological technology. Categories of service evaluation, including measures of user satisfaction, clinical outcome and financial benefit, are discussed in relation to facilitating a radiology service of the highest quality possible. PMID- 14757919 TI - Sensors in neonatal monitoring: current practice and future trends. AB - Monitoring the status of preterm infants in the Neonatal Intensive Care Unit (NICU) provides a unique and challenging environment for the design, function and use of sensor-based monitoring equipment. This article presents an overview of sensor-based instrumentation used in the NICU for physiological and chemical status monitoring, and discusses some of the key sensing principles currently in use. The clinical demand for reliable patient data at acceptable cost is driving the development of new types of monitoring technologies, in particular for continuous blood-chemistry analysis. We describe some of the new sensor-based products finding their way into the NICU, together with a review of the more promising emerging sensor technologies that might eventually be incorporated into routine neonatal monitoring practice. PMID- 14757920 TI - Continuous monitoring of tracheal pressure including spot-check of endotracheal tube resistance. AB - During mechanical ventilation, the resistance of the endotracheal and tracheostomy tube (ETT) highly influences analysis of respiratory system mechanics and imposes additional work of breathing for the spontaneously breathing patient which both can be circumvented by applying the automatic tube compensation (ATC) mode. In the ATC mode, tracheal pressure (ptrach) is continuously calculated on the basis of measured flow and airway pressure using predetermined tube specific coefficients. However, as during long-term ventilation the ETT might become partially obstructed by secretions or tube kinking, the predetermined coefficients are no longer valid rendering calculation of ptrach inaccurate. We propose an easy-to-handle maneuver for the bedside determination of current tube coefficients in the tracheally intubated patient. Based on check-spot measurement of ptrach, current tube coefficients are determined by a least-squares fit procedure valid for the partially obstructed ETT with the indwelling pressure-measuring catheter (PMC). To correct for the removal of the PMC, the relationship between tube coefficients with and those without indwelling PMC has been determined in a laboratory investigation. Accuracy of the procedure was determined during artificial ETT obstruction by comparing calculated with measured ptrach. Correspondence between calculated and measured ptrach has been found excellent. We conclude that by adopting this bedside procedure periodically, accurate calculation of ptrach is guaranteed and the advantages of the ATC mode are ensured even in long-term ventilatory support. PMID- 14757921 TI - A system for the acquisition of reproducible digital skin lesions images. AB - A major issue concerning the design and implementation of an image acquisition system for skin lesions is its ability to capture reproducible images. The reproducibility is considered essential for image analysis and for the comparison of sequential images during follow-up studies. This paper describes a prototype image acquisition system that includes a standardized illumination and capturing geometry with polarizing filters and a series of software corrections: Calibration to Black, White and Color for color constancy, Internal camera Parameters adjustment and Pose extraction for stereo vision, Shading correction and Noise Filtering for color quality. The validity of the calibration procedure and the images' reproducibility were tested by capturing sample images in three different lighting conditions: dark, medium and intense lighting. For each case the average values of the three color planes RGB and their standard deviations were calculated; the measured error differences ranged between 0.7 and 12.9 (in the 0-255 scale). Preliminary experiments for stereo measurements provided repeatability of about 0.3 mm. The above results demonstrate the reproducibility of the captured images at a satisfactory level. The developed prototype was also evaluated clinically, for its ability to support the construction of knowledge based decision systems and for telemedicine, thus to support telemedical sessions in dermatology. PMID- 14757922 TI - The hemodynamic effects of compliance, bulging, and curvature in a saphenous vein coronary artery bypass graft model. AB - The development of Intimal Hyperplasia (IH) in saphenous vein coronary artery bypass grafts (SV-CABG) is responsible for the short-term patency of these grafts. Previous studies of SV-CABG models were performed on rigid anastomotic vessels. However, the effects of compliance, bulging and curvature at the anastomosis on the general hemodynamic field, due to compliance and geometric mismatch between the vein and the artery have not been evaluated. We studied axial and transverse velocities by Laser Doppler Velocimetry on a compliant, in vitro, anatomical model of an end-to-side saphenous vein graft (SVG) to left anterior descending (LAD). The model incorporated a bulge at the sinus and curvature at the graft-host junction. Physiologic pressure and flow conditions pertaining to SV-CABG were applied. The presence of the bulge and curvature showed differences in the velocity profiles in comparison with previous rigid model studies. Dynamic separation zones were temporally augmented at the flow divider. The moving stagnation point at the floor of the host vessel was observed to move past the toe of the model during the accelerating portion of the cycle. These findings suggest that the presence of the bulge curvature and compliance may further favor conditions for the development of intimal hyperplasia (IH) at the floor of a CABG. PMID- 14757924 TI - Functional Outcome Questionnaire for Aphasia: overview and preliminary psychometric evaluation. AB - OBJECTIVE: To perform a preliminary evaluation of a questionnaire designed to assess functional outcomes of treatment for aphasia. METHODS: The Functional Outcome Questionnaire for Aphasia (FOQ-A) is an inventory consisting of 32 caregiver-completed items rating the person with stroke's ability to perform various communication behaviors on a 5-point scale. The total score for the FOQ-A is reported as a mean of all completed items. The FOQ-A was administered by research assistants to caregivers of patients with left hemisphere stroke (n = 18). RESULTS: The FOQ-A showed acceptable initial reliability and validity properties. As predicted, the FOQ-A displayed high positive correlations with standardized functional communication measures. The FOQ-A also showed only weak associations with other measures (health-related quality of life, caregiver strain). CONCLUSIONS: Overall results of our preliminary psychometric examination of the FOQ-A were promising. The findings suggested that the FOQ-A may be a sensitive measure of functional communication abilities and does not appear to be biased heavily by caregiver strain. Plans for further evaluation of the FOQ-A are discussed. PMID- 14757925 TI - Adult survivors of severe cerebral hypoxia--case series survey and comparative analysis. AB - Over 16 years, Wilson [31] saw and assessed 567 patients, 18 (3.2%) had a primary diagnosis of cerebral hypoxia. The present patient survey includes all referrals for assessment, management/advice and neuropsychological rehabilitation to a part time clinical neuropsychology service, who were seen by the first author over a five year period (October 1995-2000). Of the total patient sample (n = 168), 13 (7.7%) had incurred hypoxic damage from a variety of causes; [3] carbon monoxide poisoning (smoke inhalation), [3] cardiac arrest, [1] accidental alcohol and drug overdose, [1] near (partial) drowning, [1] near hanging (suffocation), [2] respiratory arrest following prolonged status epilepticus, [1] respiratory arrest following severe pneumonia and [1] following Addisonian crisis. The survey includes a sub-group of patients in vegetative and minimally responsive states on referral. Wilson [31] highlighted that considerable variation in cognitive functioning is likely to be observed depending on (a) nature or cause of the hypoxic insult and (b) the degree of anoxia/hypoxia experienced itself. The results of the present survey when compared with Wilson's earlier work provide a larger total data-set from which to draw conclusions and has implications for practitioners who see such patients and are involved in their multidisciplinary management and rehabilitation. PMID- 14757926 TI - Cortical remapping in amputees and dysmelic patients: a functional MRI study. AB - OBJECTIVES: To investigate motor cortex function in upper and lower limb amputees and dysmelic patients using fMRI. MATERIAL AND METHODS: Five amputees and two dysmelic patients were examined. Motor and imagery tasks were defined according to each patient limb deficiency. Cortical activation patterns were analysed for each patient and compared between groups, integrating patients clinical data. RESULTS: There is a consistent pattern of cortical reorganization in all amputees: predominance of activation in the ipsilateral motor cortex and extension to premotor and sensory areas of the contralateral cortex. On the contrary, cortical maps of dysmelic patients were similar to those of healthy volunteers, predominantly with activation of contralateral primary motor cortex areas. CONCLUSIONS: fMRI discloses specific patterns of cortical reorganization on amputees and dysmelic patients, suggesting influence by prosthesis adaptation or stump use with dexterity. These findings could be further applied in influencing neurorehabilitation and development of prosthetic devices. PMID- 14757927 TI - Cognitive impairment in patients with chronic whiplash-associated disorder--a matched control study. AB - AIM: To verify the occurrence of cognitive impairment in patients with chronic whiplash-associated disorder (WAD) and to provide a more detailed description of the impairment's character and context. METHODS: Thirty (30) patients with chronic WAD and 30 matched healthy controls completed a cognitive test battery. Four computerised tests were used: a) two different types of cognitive tasks (reaction time vs. working memory) and b) two types of information processing (verbal vs. spatial). Before testing and after every randomised subtest, subjects rated their pain level on a visual analogue scale. RESULTS: A worse overall performance among patients with WAD and, specifically, worse results concerning working memory tasks were found. Post-hoc testing revealed a statistically significant difference concerning the single variable "verbal reaction time". Pain intensity among patients increased significantly during testing. Pain intensity after the subtest for verbal mental reaction time (independent of test sequence) was significantly correlated with results in this subtest, the more pain, the more time was needed. CONCLUSION: Compared to healthy controls, patients performed worse overall. Concerning verbal reaction time, the impairment was correlated with pain intensity. The findings support the hypothesis that pain might be one important factor leading to cognitive impairment in patients with chronic WAD. PMID- 14757928 TI - Median SSEP changes in hemiplegic stroke: long-term predictive values regarding ADL recovery. AB - OBJECTIVE: To assess the predictive value of median somatosensory evoked potentials (SSEP) in the acute phase of brain infarction or hemorrhage regarding long-term prognosis. MATERIALS AND METHODS: Ninety-four stroke patients mean age 61.2, SD 11.8) were included. CT confirmed diagnoses were: cortical middle cerebral artery (MCA) infarction in 35; subcortical MCA 11; mixed 25. By size, infarctions were: massive, 29; restricted, 33; and lacunar, 9. The number of patients with thalamic hemorrhage was 8; putamenal hemorrhage, 7; other, 8. All patients presented with severe hemiparesis (54) or hemiplegia (40) with hemihipoesthesia in 89 patients. Median SSEP were recorded early (up to 7th day, mean 5.2 days, SD 0.72). SSEP parameters (presence/absence of SSEP, absolute and relative latency, amplitude of early waveforms) were compared to motor (Medical Research Council scale) and functional ability (Barthel index) at 3 and 12 months after stroke. RESULTS: Absolute N20 amplitudes and amplitude ratio evidenced almost similar predictive values that reached 66.4% at 12 months. Combined application of N20 and MRC gains provided significantly stronger prognostic information which reached 72%. CONCLUSIONS: Median SSEP parameters may serve as independent predictors of outcome. Most informative in prognosis in the early stage of stroke was the combined assessment of MRC and N20-P25 amplitude ratio. PMID- 14757929 TI - Historical approaches to the treatment of Adductor-Type Spasmodic Dysphonia (ADSD): review and tutorial. AB - Adductor-type spasmodic dysphonia (ADSD) is a voice disorder of uncertain, though likely neurogenic origin. Symptoms of the disorder include mild to profound "strain and strangle" sensations during voiced speech tasks that, in the most severe form, are physically and psychologically debilitating. Over the years, treatment approaches have evolved from behavioral attempts at voice modification, to surgical and, most recently, pharmacological treatments involving partial and temporary paralysis of muscle fibers within the larynx following injection of botulinum toxin or Botox. The most current research hints at the potential benefits of a multi-faceted approach to symptom management, combining behavioral with pharmacological treatments. The following is intended as a review and tutorial of treatment approaches for ADSD. The tutorial is intended for practicing clinical professionals with an interest in the treatment of neurogenic disorders of voice and speech. PMID- 14757930 TI - Spinal cord regeneration: moving tentatively towards new perspectives. AB - The failure of the adult human spinal cord to regenerate following injury is not absolute, but appears to be amenable to therapeutic manipulation. Recent work has shown that the provision of a growth permissive environment by the neutralization of inhibitory influences, or the grafting of fetal tissue, peripheral nerve, Schwann cells, or olfactory ensheathing cells can enhance regeneration in animal models of spinal cord injury. Stem cells are gaining ever-increasing favour as a treatment option for spinal cord injury. The potential of neural stem cells, embryonic stem cells, and bone marrow stromal cells is discussed. Additional treatment options such as pharmacological interventions, functional electrical stimulation and physiotherapy approaches are also explored. Basic science insights are used as a foundation for a discussion of a variety of clinical perspectives including repair of the chronically injured spinal cord, animal models of human spinal cord injuries and clinical trials. A more holistic approach towards spinal cord injury is suggested, one where a hierarchy of needs is recognised and quality of life is paramount. Finally, this review cautions against overly grandiose claims of an imminent miracle cure for human spinal cord injury. PMID- 14757931 TI - Transferrin C2 variant does confer a risk for Alzheimer's disease in caucasians. AB - Several gene mutations are associated with an increased risk of Alzheimer's disease. Previous studies reported higher transferrin C2 allele frequencies in Alzheimer's disease compared with normal controls. However, potential interactions between transferrin C2 and APOE (epsilon 4), have not been extensively investigated and have been the subject of controversial reports from several laboratories. We have carried out a case-control study on the association between Alzheimer's disease and transferrin C2 and APOE epsilon 4 alleles. epsilon 4 allele was associated with a four fold increase in the risk of disease, and transferrin C2 allele was significantly associated with Alzheimer's disease only in epsilon 4 negative subjects. These results suggest that apoE and transferrin may be part of a complex mechanism in the pathogenesis of Alzheimer's disease. PMID- 14757932 TI - Effects of donepezil treatment on rat nicotinic acetylcholine receptor levels in vivo and in vitro. AB - Research on acetylcholinesterase inhibitors (ChEIs) indicates that long term exposure increases the level of nicotinic acetylcholine receptors (nAChRs) but the effects of donepezil on nAChRs are not well studied. Therefore, we investigated the effects of sub-chronic donepezil administration on nAChRs in rats and rat pheochromocytoma PC-12 cells. Male Sprague Dawley rats were administered donepezil (0.7 and 2.4 micromoles/kg), nicotine (2.5 micromoles/kg) or saline subcutaneously twice daily for 14 days, PC-12 cells were incubated with 10(-6) to 10(-4) M donepezil for 72 hours and nAChR levels were determined by receptor binding assay using the nAChR ligands [3H]-epibatidine (EPI) for non alpha 7 nAChRs and [3H]-methyllyconitine (MLA) for alpha 7 nAChRs. Chronic donepezil administration at 1.4 micromoles/kg/day and 4.8 micromoles/ kg/day significantly increased [3H]-epibatidine binding in the cortex to 126 +/- 1.3% and 127 +/- 3.2% of the saline control animals, respectively. [3H]-MLA binding in the cortex increased to 114 +/- 4.4% and 124 +/- 2.8% of the control group for the high and low dose groups, respectively. Hippocampal [3H]-EPI binding in the low dose and high dose groups significantly increased to 135 +/- 3.6% and 125 +/- 4.6% of the controls, respectively while there were no changes in the level of [3H]-MLA binding. In striatal homogenates, neither [3H]-EPI nor [3H]-MLA binding were significantly effected at either dose of donepezil. In PC-12 cells, [3H]-EPI binding was increased at the non-physiological 10(-4)M concentration only. There was no effect of donepezil on [3H]-MLA binding at any concentration examined. These results indicate that donepezil increases cortical alpha 7 and non-alpha 7 nAChRs, hippocampal non-alpha 7 nAChRs but does not influence striatal nAChR levels. Furthermore, the lack of an effect on the alpha 7-nAChRs in PC-12 cells suggests that the increase in cortical alpha 7 nAChRs may be an indirect effect of increased acetylcholine levels in vivo. PMID- 14757933 TI - The effect of chronic aluminum(III) administration on the nervous system of aged rats: clues to understand its suggested role in Alzheimer's disease. AB - The effect of chronic aluminum intake has been investigated in the brain of aged male Wistar rats to assess the potential role of the accumulation of this metal ion on the development of neurodegenerative features observed in Alzheimer's disease. AlCl3 x 6 H2O (2g/L) was administered to experimental animals for 6 months in the drinking water. The total content of Al (microg/g fresh tissue) was measured by inductively coupled plasma atomic emission spectrometry (ICP-AES), while the content of Cu, Zn and Mn was determined by flame AAS in the prosencephalon + mesencephalon, pons-medulla and cerebellum of control and Al(III)-treated animals. The area occupied by mossy fibres in the CA3 field of the hippocampus was estimated by a computer-assisted morphometric method following Timm's preferential staining. In Al(III)-treated rats the concentration of Cu, Zn and Mn did not increase significantly (p < 0.5) in prosencephalon + mesencephalon, nor in pons-medulla (p < 0.5) except for Cu (p < 0.05) in pons medulla. In the cerebellum the only significant increase was seen for Zn (p < 0.01) while no change was observed for Cu and Mn. The area occupied by the mossy fibres in the hippocampal CA3 field was significantly increased (+32%) in aged Al(III)-treated rats. Since Cu, Zn and Mn are essential components of the cytosolic and mitochondrial superoxide dismutases, it is possible that the increased content of these ions in aged Al(III)-treated rats represents an increased amount of genetic expression of these antioxidant enzymes. Considering that the positivity to Timm's reaction is based on the presence of free or loosely bound Zn2+ ions within synaptic terminals and that Zn2+ ions are reported to be accumulated by hippocampal neurons when tissue injury occurs, the increased area of the mossy fibres in CA3 field of Al(III)-treated rats could indicate increased hippocampal damage in these animals. Taken together, the present findings indicate that the aging CNS is particularly susceptible to Al(III) toxic effects which may increase the cell load of oxidative stress and may contribute, as an aggravating factor, to the development of neurodegenerative events as observed in Alzheimer's disease. PMID- 14757934 TI - Ubiquitin-negative mini-pick-like bodies in the dentate gyrus in p301l tauopathy. AB - Neuropathological and biochemical findings are reported in a patient who had suffered from frontotemporal dementia associated with a P310L mutation in the tau gene and included in the H1 haplotype. Tau accumulation, as revealed with phospho specific anti-tau antibodies Thr181, Ser199, Ser202, Ser214, Ser262, Ser396, Ser422 and AT8 (Ser202 and Thr205), was found in neurons with pre-tangles, and astrocytes and oligodendrocytes through the brain. The most characteristic feature was tau immunoreactivity decorating the perinuclear region and small cytoplasmic aggregates designed as mini-Pick-like bodies, mainly in the dentate gyrus. Inclusions were not stained with anti-ubiquitin antibodies and did not recruit tubulins. Tau accumulation in individual cells was associated with increased expression of kinases linked with tau phosphorylation, mainly active (phosphorylated) stress kinases SAPK/JNK and p38 (SAPK/JNK-P and p38-P). Phosphorylated GSK-3 beta at Ser9 (GSK-3 beta-P), that inactivates the kinase, was particularly abundant in mini-Pick-like bodies, thus suggesting alternative roles of GSK-3 probably involved in cell survival. Western blots of sarkosyl insoluble fractions revealed a double band pattern of phospho-tau of 68/66 kDa and 64 kDa in the hippocampus and white matter in the P310L mutation. Sarkosyl insoluble fractions of the hippocampus were enriched in p38-P and GSK-3 beta-P in Alzheimer's disease (AD) cases, processed in parallel for comparative purposes, but not in the P310L mutation. In addition, no bands of high molecular weight were found in P310L in contrast with AD in these fractions. These findings indicate that the major sites of tau phosphorylation, and the expression of kinases involved in tau phosphorylation are active in P310L mutation as in AD and other tauopathies. Yet the P310L mutation has particular phospho-tau inclusions that are not tag with ubiquitin and appear to be rather soluble when compared with AD. PMID- 14757935 TI - Reduced expression of the cystic fibrosis transmembrane conductance regulator gene in the hypothalamus of patients with Alzheimer's disease. AB - BACKGROUND: The cystic fibrosis transmembrane conductance regulator (CFTR) gene encodes an approximately 150-165 kD glycoprotein that is mutated in individuals with cystic fibrosis. Previous studies demonstrated expression of the CFTR gene in the hypothalamus, suggesting a potential role for this molecule in the regulation of systemic metabolic functions. Individuals with cystic fibrosis often exhibit wasting and marked reductions in body fat content. Since the hypothalamus is a late target of neurodegeneration in Alzheimer's disease (AD), we postulated that patients with end-stage AD and bodily wasting would have reduced levels of CFTR expression in the hypothalamus. METHODS: CFTR mRNA and protein were examined in postmortem hypothalamic tissue from 11 AD and 7 aged controls using in situ hybridization and immunohistochemical staining. Standardized sections that included the supra-optic, paraventricular, anterior, and ventromedial nuclei, and the lateral hypothalamus were studied. RESULTS: The density of CFTR+ neurons and the intensity of the CFTR hybridization signals were strikingly reduced in AD. Immunohistochemical staining studies demonstrated CFTR immunoreactivity most prominently distributed in small clusters of neurites (5 to 20 in number). Digital image quantification showed that the density of CFTR+ neurites was significantly reduced in AD relative to aged control samples (P=0.001). However, there was no evidence for selective involvement of particular hypothalamic nuclei. CONCLUSIONS: CFTR gene expression is down-regulated and its corresponding immunoreactivity reduced in AD relative to control hypothalamic tissue. Reduced CFTR expression in the hypothalamus may represent an important mechanism by which AD neurodegeneration contributes to body wasting in the late stages of disease. PMID- 14757936 TI - The clinical diagnosis of Alzheimer's disease without the use of head imaging studies. A cliniconeuropathological study. AB - Although head imaging studies are frequently used in the work-up of dementia, published criteria for the clinical diagnosis of Alzheimer's disease (AD) do not require them. Since our brain bank contains cases in which physicians had specifically diagnosed AD without using a head imaging study, we thought it of interest to investigate the accuracy of their clinical diagnoses. We retrospectively reviewed 911 consecutive dementia cases for those clinically diagnosed as either AD or senile dementia (SD). Twenty-one were identified in which head imaging studies had not been used, each diagnosed as AD or SD by a different physician. In only three had the physician reported a reason why a study was not done. In all 21 cases the primary neuropathological cause of the dementia was AD. Neuropathology in addition to AD was also noted, including cortical Lewy bodies in three, infarcts on gross examination in three, multiple microscopic infarcts in four, and multiple cerebral metastases in one. Acknowledging a number of study limitations, it is remarkable that the judgment of the physicians was correct regarding AD in all 21 cases. It is questionable if a head CT or MRI scan at time of diagnosis would have benefited any of the patients. PMID- 14757937 TI - Advanced glycation endproduct precursor alters intracellular amyloid-beta/A beta PP carboxy-terminal fragment aggregation and cytotoxicity. AB - Carbonyl stress from products of lipid peroxidation, such as 4-hydroxynonenal (HNE), and products of sugars in diabetes mellitus, such as methylglyoxal (MG) and glyoxal (G), may contribute to neurodegeneration in Alzheimer's disease (AD). We tested the hypothesis that these carbonyls alter the proposed central pathogenic mechanism of AD, intracellular amyloid-beta (A beta)-mediated cytotoxicity, using a human neuroblastoma cell line that conditionally expresses carboxy-terminal fragments (CTFs) of the amyloid precursor protein. HNE was a potent cytotoxin, whereas G was mildly cytotoxic; cytotoxicity from each was independent of A beta/CTF expression and not altered by alpha-tocopherol. In contrast, MG cytotoxicity was enhanced by the induced expression of A beta/CTFs and suppressed by alpha-tocopherol. alpha-tocopherol cytoprotection was accompanied by decreased A beta/CTF aggregation. G also promoted beta/CTF aggregation but by mechanisms unaffected by alpha-tocopherol treatment. Our findings showed that A beta/CTF aggregation and cytotoxicity may be profoundly altered by aldehydes associated with diabetes and that in the case of MG, this process is suppressed by alpha-tocopherol. Moreover, our results suggest that while intracellular aggregation of A beta/CTFs may be necessary for the development of toxicity attributable to their expression in this model, the presence of high-molecular weight aggregated A beta/CTFs does not invariably lead to cytotoxicity. PMID- 14757938 TI - A case history illustrating how extended release cholinesterase inhibitors could improve management of Alzheimer's disease. PMID- 14757939 TI - Cerebral atherosclerosis and oxidative stress in Alzheimer's disease. PMID- 14757941 TI - Biomarker amplification by serum carrier protein binding. AB - Mass spectroscopic analysis of the low molecular mass (LMM) range of the serum/plasma proteome is a rapidly emerging frontier for biomarker discovery. This study examined the proportion of LMM biomarkers, which are bound to circulating carrier proteins. Mass spectroscopic analysis of human serum following molecular mass fractionation, demonstrated that the majority of LMM biomarkers exist bound to carrier proteins. Moreover, the pattern of LMM biomarkers bound specifically to albumin is distinct from those bound to non albumin carriers. Prominent SELDI-TOF ionic species (m/z 6631.7043) identified to correlate with the presence of ovarian cancer were amplified by albumin capture. Several insights emerged: a) Accumulation of LMM biomarkers on circulating carrier proteins greatly amplifies the total serum/plasma concentration of the measurable biomarker, b) The total serum/plasma biomarker concentration is largely determined by the carrier protein clearance rate, not the unbound biomarker clearance rate itself, and c) Examination of the LMM species bound to a specific carrier protein may contain important diagnostic information. These findings shift the focus of biomarker detection to the carrier protein and its biomarker content. PMID- 14757942 TI - p53 and erbB-2 are not associated in matched cases of primary and metastatic ovarian carcinomas. AB - Ovarian cancer has a high mortality rate largely due to the limited number of ovarian carcinomas detected at an early stage. Understanding the molecular changes occurring during the progression of ovarian carcinoma would aid in the development of therapies that may inhibit or target metastasis. Primary and metastatic lesions from 54 and 40 patients with advanced ovarian carcinoma, respectively (including matched primary and metastatic lesions from 30 patients) were evaluated for nuclear accumulation of p53 (clone BP53-12-1) and cytoplasmic and membranous immunostaining of p185 erbB-2 (clone 3B5) by immunohistochemistry. No differences in the immunostaining of p53 and p185erbB-2 (cytoplasm or membrane) were observed between primary and metastatic lesions of the matched cases. Similarly, no differences in the proportion of positive cases of p53 between primary and metastatic lesions of the matched cases was observed. Thus, novel therapies that target p53 or p185erbB-2 can utilize specimens from either primary or metastatic lesions to characterize these targets prior to therapy. Spearman correlations between p53 and p185erbB-2 (cytoplasm or membrane) immunohistochemistry scores were insignificant for the matched cases, all primary lesions, and all metastatic lesions. Also, no significant associations occurred between nuclear accumulation of p53 (positive versus negative) and phenotypic expression of p185erbB-2 (cytoplasm or membrane) immunostaining scores for the matched cases, all primary lesions, and all metastatic lesions. Thus, the nuclear accumulation of p53 and immunostaining of p185erbB-2 in the cytoplasm or on the cellular membranes are independent. PMID- 14757943 TI - Peroxisomal enzymes and 8-hydroxydeoxyguanosine in rat liver treated with perfluorooctanoic acid. AB - Although peroxisome proliferators are considered non-genotoxic agents, most of them, nevertheless, were found to promote and/or induce, hepatocellular carcinoma (HCC) in rodents. The aim of the present study is, first, to investigate whether the peroxisome proliferator perfluorooctanoic acid (PFOA) possesses inherent liver cancer promoting activity, and second, to study the possible mechanisms involved. To acheive these aims two protocols have been applied, a biphasic protocol (initiation by diethyl-nitrozamine (DEN) 200 mg/kg i.p. followed by treatment with 0.005% or 0.02% perflourooctanoic acid (PFOA) for 14 and 25 weeks) and a triphasic initiation, selection-promotion (IS) protocol (initiation by giving 200 mg/kg DEN i.p. followed by a selection procedure for 2 weeks consisting of giving 0.03% 2-acetylaminofluorene (2-AAF) in diet). In the middle of this treatment a single oral dose of carbon tetrachloride (2.0 ml/kg) was given, followed by giving diet containg 0.015% of PFOA for 25 weeks. After applying both protocols, our results showed slight increase in the catalase activity while acyl CoA oxidase activity was markedly increased. Both experiments indicated that PFOA has a liver cancer promoting activity. Other groups of rats were given either basal diet or diet containing 0.02% PFOA. Five or nine weeks later they were sacrificed and the levels of 8-hydroxydeoxyguanosine in the isolated DNA were estimated. The data showed a slight nonetheless insignificant increase in 8-hydroxydeoxyguanosine. From the present data, it is concluded that PFOA is a true liver cancer promoter that may not require extensive initial DNA damage for its promoting activity. PMID- 14757944 TI - Plasma homocysteine and oxidative stress in cardiovascular disease. AB - Hyperhomocysteinemia (Hhcy) has been associated with pathological and stressful conditions and is a risk factor for cardiovascular disease (CVD). The aim of this study was to evaluate the correlation between plasma homocysteine (hcy) and lipid peroxidation in patient with CVD. This study was carried out on 40 patients with CVD as well as 15 healthy volunteers of comparable age and gender as control group. The patients were divided into 2 groups as follows: group I, included 20 patients with acute myocardial infarction and group II, included 20 patients with atherosclerotic coronary artery disease with no evidence of previous myocardial infarction. Plasma hcy, nitric oxide (NO) and malondialdhyde (MDA) [as index of lipid peroxidation] were measured in all groups. In addition serum total cholesterol, HDL, LDL and triglycerides were evaluated. Results obtained showed that, there was a significant elevation in the levels of plasma hcy, NO and MDA in groups I and II as compared to control group. There was a strong positive correlation between plasma hcy and MDA (r = 0.59, p < 0.001). Also NO was positively correlated with both hcy (r = 0.49, p < 0.001) and MDA (r = 0.51, p < 0.001). Serum total cholesterol, LDL, and triglycerids were also significantly elevated while serum HDL was significantly decreased in groups I and II as compared to control group. It can be concluded that, hyperhomocysteinemia is a possible factor in free radical generation and therefore cardiovascular diseases. PMID- 14757945 TI - Development of proteomic patterns for detecting lung cancer. AB - Lung cancer is at present the number one cause of cancer death and no biomarker is available to detect early lung cancer in serum samples so far. The objective of this study is to find specific biomarkers for detection of lung cancer using Surface Enhanced Laser Desorption/Ionization (SELDI) technology. In this study, serum samples from 30 lung cancer patients and 51 age-and sex-matched healthy were analyzed by SELDI based ProteinChip reader, PBSII-C. The spectra were generated on WCX2 chips and protein peaks clustering and classification analyses were performed utilizing Biomarker Wizard and Biomarker Patterns software packages, respectively. Three protein peaks were automatically chosen for the system training and the development of a decision classification tree. The constructed model was then used to test an independent set of masked serum samples from 15 lung cancer patients and 31 healthy individuals. The analysis yielded a sensitivity of 93.3%, and a specificity of 96.7%. These results suggest that the serum is a capable resource for detection of specific lung cancer biomarkers. SELDI technique combined with an artificial intelligence classification algorithm can both facilitate the discovery of better biomarkers for lung cancer and provide a useful tool for molecular diagnosis in future. PMID- 14757947 TI - Investigation into the material properties of beech wood and cortical bone. AB - When testing medical implants it is very important to be able to test the implant using a suitable material. In the case of orthopaedic implants the optimum material is bone. Beech wood is considered a suitable substitute for bone as it has a similar Young's modulus in tension. Although it is widely used, no actual comparison of the two materials has been undertaken. The aim of this study was to compare the material properties of beech wood and cortical bone using conventional compression tests. Cortical bone samples 4 mm in diameter and 20 mm in length, were prepared from the tibia of an amputated leg. Beech wood samples were prepared to the same specifications. In compression, the Young's modulus for cortical bone was found to be 27+/-9.9 GPa (mean +/- standard deviation) and for beech wood 2.6+/-1.7 GPa. The failure load for cortical bone was 911+/-207 N and 732+/-62 N for beech wood. Although beech wood has been used as a substitute for bone in some studies, this study has shown that there are significant differences in the properties of the two materials when they are subjected to compression. PMID- 14757946 TI - Haplotype analysis of Norwegian and Swedish patients with acute intermittent porphyria (AIP): Extreme haplotype heterogeneity for the mutation R116W. AB - Acute intermittent porphyria (AIP), the most common of the acute porphyrias, is caused by mutations in the gene encoding hydroxymethylbilane synthase (HMBS) also called porphobilinogen deaminase (PBGD). The mutation spectrum in the HMBS gene is characterized by a majority of family specific mutations. Among the exceptions are R116W and W198X, with high prevalence in both the Dutch and Swedish populations. These two mutations were also detected in unrelated Norwegian patients. Thus, Norwegian and Swedish patients were haplotyped using closely linked flanking microsatellites and intragenic single nucleotide polymorphisms (SNPs) to see if the high frequency of these two mutations is due to a founder effect. Twelve intragenic SNPs were determined by a method based on fluorescent restriction enzyme fingerprinting single-strand conformation polymorphism (F-REF SSCP). W198X occurred exclusively on one haplotype in both Norwegian and Swedish patients, showing that it has originated from a common gene source. In contrast, R116W was found on three different haplotypes in three Norwegian families, and in five Swedish families on four or five haplotypes. This extreme haplotype heterogeneity indicates that R116W is a recurrent mutation, maybe explained by the high mutability of CpG dinucleotides. This can also explain why it is the only AIP mutation reported to occur in seven different populations (Norway, Sweden, Finland, Netherlands, France, Spain and South Africa). PMID- 14757949 TI - Comparison of mechanical properties of rat tibialis anterior tendon evaluated using two different approaches. AB - Tendon injuries may result in variations of its mechanical properties. The published data of the tendon stiffness of small animals, such as mouse and rat, are exclusively obtained by measuring grip-to-grip (g-t-g) displacement. Local strain concentration and relative sliding of the specimens in the clamps might significantly affect the measured tendon deformation. In the present study, the mechanical properties of the rat tibialis anterior tendon measured using the proposed tendon mark method were compared to those evaluated using the g-t-g displacement method. Five male Sprague Dawley rats ( approximately 418 g) were used in this study. For the proposed method, reference marks were made on the tendons using permanent ink. A microscope video system was customized to observe and record the tendon deformation. Pattern recognition software was developed to obtain the displacement time-histories of the reference marks. The distance between the grips was approximately 7 mm; and the distance between the reference marks used for the data processing was approximately 5 mm. The cross-section areas of the specimens were measured using a custom-made slot gauge and by applying a constant compressive stress (0.15 MPa). The tendons were clamped between two custom-made metal grips and stretched on a testing machine at a constant speed (1 mm/s) up to failure. Throughout the tests, the tendon specimens were submerged in a PBS bath at 22 degrees C. The deformation of the specimens was evaluated using the g-t-g displacement method and the proposed method. The stress/strain curves obtained by using the g-t-g displacement can be characterized by an initial toe zone, a quasi-linear zone, and a final failure stage. The stress/strain curves determined using the proposed method are quite different from those obtained using the g-t-g displacement: it has a smaller toe zone and a stress-hardening transition, over which the tendon stiffness increases dramatically with the increasing strain. The tendon stiffness measured by using the g-t-g displacement method may underestimate the actual mechanical properties of tendon by approximately 43%. PMID- 14757948 TI - Mechanism of apatite formation on pure titanium treated with alkaline solution. AB - The mechanism of bone-like apatite formation on the surface of pure titanium pretreated with NaOH solution is still being investigated. The apatite formation may depend on the solution that is used. In the present study, several types of solutions such as simulated body fluid (SBF), calcium aqueous solution (CAS), and phosphate aqueous solution (PAS) were used to investigate bone-like apatite formation on alkali-treated titanium. In order to observe the effect of hydrolysis on the apatite formation, experiments of pretreated titanium immersed in distilled water before the immersion in SBF were also conducted. The results showed that the mechanism of apatite formation was the hydrolysis reaction of sodium titanate which induced the apatite formation. The pre-precipitation of either calcium or phosphate could prevent the apatite formation on the surface of alkaline treated titanium. PMID- 14757950 TI - Counter-propagation network with variable degree variable step size LMS for single switch typing recognition. AB - Morse code is now being harnessed for use in rehabilitation applications of augmentative-alternative communication and assistive technology, including mobility, environmental control and adapted worksite access. In this paper, Morse code is selected as a communication adaptive device for disabled persons who suffer from muscle atrophy, cerebral palsy or other severe handicaps. A stable typing rate is strictly required for Morse code to be effective as a communication tool. This restriction is a major hindrance. Therefore, a switch adaptive automatic recognition method with a high recognition rate is needed. The proposed system combines counter-propagation networks with a variable degree variable step size LMS algorithm. It is divided into five stages: space recognition, tone recognition, learning process, adaptive processing, and character recognition. Statistical analyses demonstrated that the proposed method elicited a better recognition rate in comparison to alternative methods in the literature. PMID- 14757951 TI - The influence of the viscosity classification of an acrylic bone cement on its in vitro fatigue performance. AB - The goal of the present work was to investigate the influence of the viscosity classification of an acrylic bone cement on its in vitro fatigue performance, as determined in fully-reversed tension-compression (+/-15 MPa) fatigue tests. The test matrix comprised six commercially available bone cements [Orthoset1, (OS1), Orthoset(R)3 (OS3), CemexRX (CRX), Cemex XL (CXL), Palacos R (PR) and Osteopal (OP)], two methods of mixing the cement constituents (hand-mixing and vacuum mixing), two methods of fabricating the test specimens (direct molding and molding followed by machining), two specimen cross-sectional shapes (rectangular or "flat" and circular or "round"), and four test frequencies (1, 2, 5, and 10 Hz). In total, 185 specimens, distributed among 20 sets, were tested. The test results (number of fatigue stress cycles, N_f) were processed using the linearized transformation of the three-parameter Weibull distribution, whence estimates of the Weibull mean, N_[WM], were obtained. Statistical analysis of the ln N_f results (Mann-Whitney test; alpha<0.05) and a comparison of the N_[WM] estimates for specimen sets in which the formulations have essentially the same composition but different viscosity classification (namely, OS1 versus OS3, CRX versus CXL, and PR versus OP) showed that, in the majority of the comparisons carried out, the viscosity classification of a bone cement does not exert a significant influence on its in vitro fatigue performance. PMID- 14757952 TI - Geometric element analysis of fretting in a model of a modular femoral component of a hip implant. AB - The study utilized the geometric element analysis method to investigate the effect of three variables (head diameter, d; head material; and neck material) on the y-direction displacement, due to fretting (delta), in a model of the head neck connection in a modular version of the femoral component of a hip implant, subjected to a resultant hip joint force only. It was found that while d and neck material exerted a strong effect on delta, the head material had practically no such effect. The implications of this finding for design and materials selection issues for this connection are discussed. PMID- 14757953 TI - Effect of exercise on the bone strength, bone mineral density, and metal content in rat femurs. AB - We have investigated the effect of exercise on the bone strength, bone mineral density (BMD), and metal content in rat femurs. Five Wister rats acting as an exercised group (E group) were exercised at intervals on a treadmill every day for four weeks. Another five rats were fed in a cage, acting as a control group (C group), without any exercise. After four weeks, the bend strength, Young's modulus, BMD value, and metal content of the femurs were measured using a three point bend test, Dual-Energy X-ray Absorptiometry (DEXA), and chemical analysis. The bend strength of the E group was 101+/-5 MPa, and that of the C group was 86+/-7 MPa. The Young's modulus was 1.16+/-0.03 GPa for the E group, and 0.98+/ 0.06 GPa for the C group. The BMD values were 0.179+/-0.002 g/cm(2) for the E group, and 0.166+/-0.002 g/cm(2) for the C group. The Ca concentration was 230600 +/- 1500 microg/g for the E group, and 223000 +/- 2700 microg/g for the C group. The Zn concentration was 160 +/- 7 microg/g for the E group, and 188 +/- 8 microg/g for the C group. PMID- 14757954 TI - Biological evaluation of hydroxyapatite/poly-L-lactide (HAp/PLLA) composite biomaterials with poly-L-lactide of different molecular weights intraperitoneally implanted into mice. AB - Histopathologic analysis of the tissue with HAp/PLLA implants was made and the leukocyte formula and chemiluminescence response of peritoneal phagocytes 2, 7 and 12 weeks after intraperitoneal implantation studied. Implants were made of HAp/PLLA biocomposites with PLLA molecular weights of 50000 (HAp/PLLA(50)) and 430000 g/mol (HAp/PLLA(430)) and of crushed devitalized femur bone of a young Wistar rat. Leukocyte formula and chemiluminescence of peritoneal phagocytes showed no systemic inflammatory response. The studied implants caused locally weak inflammatory reaction. The resorption of implants ranges in intensity (polymer resorption, i.e. disappearance rate), from the highest with the bone implants, low with HAp/PLLA(50), to the lowest with the HAp/PLLA(430) implants. Good resorption of the biocomposites and its mutual ingrowth with connective tissue prove their good biocompatibility. PMID- 14757955 TI - A comparison of stainless steel and CP titanium rods for the anterior instrumentation of scoliosis. AB - Use of 4.0 mm and 5.0 mm steel rods have resulted in proximal screw pullout. Titanium rods, which encompass a reduced yield point, may increase the effective stiffness of the construct when used with segmental anchors. Seven human thoracic spines were loaded in axial compression, axial torsion, flexion, extension, and lateral bending. Testing was performed on intact, discectomy and simulated bony fusion specimens. Specimens were randomly instrumented with 4.0 mm and 5.0 mm steel, and 4.5 mm CP Ti rods. In compression and torsion, the intact spine demonstrated increased stiffness with respect to all instrumentation employed in a discectomy condition. No significant differences between the intact and the instrumentation systems were detected for torsion or compression under simulation of fusion. Under flexural loading, no significant differences were detected between the intact specimen and the instrumentation systems. When used in conjunction with segmental force anchors, the use of CPTi rods which posses reduced yield points can provide sufficient rigidity as compared to stainless steel rods of comparable diameter. Implants of reduced yield point will permit permanent deformation with low force. Screw pullout may result when high yield materials are employed in conjunction with anchors. PMID- 14757956 TI - Ultra high molecular weight polyethylene (UHMWPE) model can provide results comparable to cadaveric models. AB - The in vitro biomechanical models using a cadaveric spine specimen have long been used in understanding normal and abnormal functions of spines as well as for strength and stability testing of the spine specimen or spinal construct. Little effort has been made to describe the similarities or differences between UHMWPE and cadaveric models. Eight cadaveric lumbar spines were harvested generating six FSU and three corpectomy models. Six UHMWPE blocks were fabricated to form FSU and corpectomy models. All were tested intact, with posterior instrumentation, and with anterior instrumentation consisting of Moss-Miami 4.0 mm stainless steel rods, uni-axial stainless steel screws and DePuy Harm's cages. All models were tested in axial compression. The cadaveric model and UHMPWE model yielded axial stiffness values of comparable magnitude with respect to instrumentation applied using the posterior approach (P>0.05). Under an FSU configuration, only in the case of anterior instrumentation without the addition of a Harm's cage did both the cadaveric and UHMPWE models provide comparable axial stiffness results (P>0.05). While in vitro cadaveric models are considered the gold standard for biomechanical testing of the spine, the data suggests that under specific approaches and surgical models UHMWPE can be used to infer mechanical performance of instrumentation in cadaveric material. PMID- 14757957 TI - Hydroxyapatite/PMMA composites as bone cements. AB - Currently PMMA is the polymer most commonly used as a bone cement for the fixation of total hip prostheses. Ideally, a bone cement material should be easy to handle, biologically compatible, nonsupporting of oral microbial growth, available in the particulate and molded forms, easy to obtain, nonallergenic, adaptable to a broad range of dental and medical applications, in possession of high compressive strength, and effective in guided tissue regenerative procedures. One of the problems associated with the conventional types of bone cement used is their unsatisfactory mechanical and exothermic reaction properties. The purpose of this in vitro study was to investigate and compare the mechanical properties (three-point bending strength, energy-to-break, and modulus of elasticity) and physical properties (setting time, water sorption, and exothermic heat) of HA/PMMA (HA group) and bovine-bone originated HA/PMMA (BB group) composites. Composites samples were fabricated by admixing method. It was found that the addition of HA and BB particles increased the water sorption. Generally 10 v/o 20 v/o HA and 0 v/o to 10 v/o BB ratio combinations had significant beneficial effects on the mechanical properties. The heat generated during polymerization was influenced by the different admixtures. More than 40 v/o HA and 40 v/o BB should be mixed into PMMA to reduce the peak temperature. Overall evaluation indicated that the BB group had better properties than the HA group. PMID- 14757958 TI - Mechanical properties of four methylmethacrylate-based resins for provisional fixed restorations. AB - The use of a provisional restoration is an important phase in the treatment of the dental prosthetic patient. A good provisional restoration should satisfy the following requirements: pulpal protection, positional stability, ease in cleaning, accurate margins, wear resistance, dimensional stability, and serve as a diagnostic aid in treatment assessment and esthetics. There is a tendency for discoloration, occlusal wear, and fracture that eventually leads to unnecessary repair. Heat-processed and reinforced methacrylate-based resins have been used to improve the mechanical and physical properties of provisional restorations. Among various improvements, the interpenetrating network crosslinked PMMA (IPN) has been shown to have superior mechanical properties if manufactured through a dough compression molding process at 130 degrees C. However, there have been no published data that relate with the use of this material for fixed provisional restorations. The objective of this study was to compare four methyl methacrylate based resins for provisional crowns and bridges with varying processing cycles, including JET [self-cure], ACRALON [heat-cured], titanium dioxide filled PMMA [heat-cured], and IPN [heat-cured denture tooth resin]. Properties studied included transverse strength, toughness, rigidity, and hardness. From the results of this study the following conclusions can be made: the IPN group may have had a lower degree of conversion as demonstrated by decreased strength, toughness, and hardness data as compared with Acralon. Increasing the polymerization cycle of unmodified Acralon resin causes a significant increase in strength. PMID- 14757959 TI - The influence of lateral electronic disequilibrium on the radiation treatment planning for lung cancer irradiation. AB - Using higher energy photons can obtain better target dose uniformity and skin sparing for treating deep lesions, but the effect of lacking lateral scattering in the low-density lung may degrade the target coverage. To analyze the influence of lateral electronic disequilibrium on the radiation treatment planning for lung cancer, three dimension conformal treatment (3D-CRT) plans of using 6 MV and 18 MV X-ray respectively for a lung cancer case have been worked out by using pencil beam algorithm and collapsed cone algorithm provided by Helax-TMS treatment planning system for the same radiation field arrangement for both energies. Dose volume histogram (DVH) in target and organs at risk (OARs) are used for comparison of different plans. The study shows that using pencil beam algorithm, the target DVH are similar for 6 MV and 18 MV plan. However, using collapsed cone algorithm that can make account of lateral electron scattering, the target is underdosed. The change is even more pronounced for 18 MV plan. The doses for lung and spinal cord are similar for these two energies and two algorithms. Therefore, for lung cancer, dose calculation algorithm should have the ability of handling accurately the effect of the tissue density heterogeneity. It is better to use the lower-energy photons (6 MV) than to use the higher-energy photons (18 MV). PMID- 14757960 TI - Thiocyanate overload and thyroid disease. AB - Thiocyanate [SCN-] is a complex anion which is a potent inhibitor of iodide transport. It is the detoxification product of cyanide and can easily be measured in body fluids. Consumption of naturally occurring goitrogens, certain environmental toxins and cigarette smoke can significantly increase SCN- concentrations to levels potentially capable of affecting the thyroid gland. Goiter endemics were reported to develop when the critical urinary iodine/ SCN- ratio decreases below 3 microg iodine per mg SCN-. Iodine supplementation completely reverses the goitrogenic influence of SCN-. SCN- is also generated from cigarette smoking as a detoxifying product of cyanide. During the past two decades many reports dealt with the possible effects of cigarette smoking on thyroid hormone synthesis, thyroid gland size and thyroid autoimmunity including infiltrative ophtalmopathy of Graves' disease. In this mini-review, issues regarding thiocyanate overload and thyroid disease will be summarized. PMID- 14757961 TI - Flavonoids and thyroid disease. AB - The most potent natural plant-derived compounds that can affect thyroid function, thyroid hormone secretion and availability to tissues is the group of flavonoids, i.e. plant pigments. They are present in our daily food, such as vegetables, fruits, grains, nuts, wine, and tea. Epidemiological studies suggest beneficial effects on health of flavonoids, which are commonly attributed to their activity as antioxidants. Experimental studies in vitro, however, showed inhibition of organification in thyroid cells and follicles by several flavonoids. Studies in vivo and vitro with synthetic and natural flavonoids showed displacement of T4 from transthyretin leading to disturbances in thyroid hormone availability in tissues. Radioactive labeled flavonoids appeared to be eliminated rapidly from the body mainly through excretion in the feces. In pregnant rats synthetic flavonoids cross the placenta and accumulate in the fetal compartment, including the fetal brain. Therefore, a high intake of flavonoids is contraindicated. IN CONCLUSION: flavonoids show strong interference with many aspects of thyroid hormone synthesis and availability. PMID- 14757962 TI - Role of iodine in antioxidant defence in thyroid and breast disease. AB - The role played in thyroid hormonogenesis by iodide oxidation to iodine (organification) is well established. Iodine deficiency may produce conditions of oxidative stress with high TSH producing a level of H_2O_2, which because of lack of iodide is not being used to form thyroid hormones. The cytotoxic actions of excess iodide in thyroid cells may depend on the formation of free radicals and can be attributed to both necrotic and apoptotic mechanisms with necrosis predominating in goiter development and apoptosis during iodide induced involution. These cytotoxic effects appear to depend on the status of antioxidative enzymes and may only be evident in conditions of selenium deficiency where the activity of selenium containing antioxidative enzymes is impaired. Less compelling evidence exists of a role for iodide as an antioxidant in the breast. However the Japanese experience may indicate a protective effect against breast cancer for an iodine rich seaweed containing diet. Similarly thyroid autoimmunity may also be associated with improved prognosis. Whether this phenomenon is breast specific and its possible relationship to iodine or selenium status awaits resolution. PMID- 14757963 TI - Vitamin E and thyroid disease: a potential link that kindles hope. PMID- 14757964 TI - Selenium and thyroid function in infants, children and adolescents. AB - Selenium is an integral component of the enzymes glutathione peroxidase (GPx) and iodothyronine deiodinases. Although selenium nutrition could conceivably affect thyroid function in infants, children and adolescents, available data suggest that the effect of selenium deficiency on thyroid function is relatively modest. In patients with isolated selenium deficiency (such as patients with phenylketonuria receiving a low-protein diet), peripheral thyroid hormone metabolism is impaired but there are no changes in thyrotropin (TSH) or clinical signs of hypothyroidism, suggesting that these patients are euthyroid. Selenium supplementation may be advisable to optimize tissue GPx activity and prevent potential oxidative stress damage. In areas where combined selenium and iodine deficiencies are present (such as endemic goiter areas in Central Africa), selenium deficiency may be responsible for the destruction of the thyroid gland in myxoedematous cretins but may also play a protective role by mitigating fetal hypothyroidism. In these areas, selenium supplementation should only be advocated at the same time or after iodine supplementation. In patients with absent or decreased production of thyroid hormones and who rely solely on deiodination of exogenous L-thyroxine for generation of the active triiodothyronine (such as patients with congenital hypothyroidism), selenium supplementation may optimize thyroid hormone feedback at the pituitary level and decrease stimulation of the residual thyroid tissue. PMID- 14757965 TI - Humic substances in drinking water and the epidemiology of thyroid disease. AB - Thyroid diseases are common in all populations but the type and frequency depends on environmental factors. In Denmark geographical differences in iodine intake are caused by different iodine contents of drinking water, which varies from < 1 to 139 microg iodine per litre. Comparative epidemiologic studies have demonstrated considerable differences in type and occurrence of thyroid disease with more goitre and hyperthyroidism in Aalborg with water iodine content around 5 microg/L, and more hypothyroidism in Copenhagen with water iodine around 20 microg/L. In Denmark, iodine in ground water is bound in humic substances, which have probably leached from marine sediments in the aquifers. Interestingly, humic substances in water from other parts of the world have goitrogenic properties, especially humic substances from coal and shale. Humic substances are heterogeneous mixtures of naturally occurring molecules, produced by decomposition of plant and animal tissues. The effect of humic substances in drinking water on the epidemiology of thyroid disease probably depends on the source of aquifer sediments. PMID- 14757966 TI - Oxidative stress and Graves' ophthalmopathy: in vitro studies and therapeutic implications. AB - Graves' ophthalmopathy (GO) is a disorder of autoimmune origin caused by a complex interplay of endogenous and environmental factors. After recognition of one or more antigens shared by thyroid and orbit, activated T lymphocytes infiltrating the orbit trigger a cascade of events leading to production of cytokines, growth factors and oxygen reactive species. Proliferation of adipocytes and fibroblasts then follows, with an increased synthesis of glycosaminoglycans (GAG), which attract water and cause edema of orbital structures and venous congestion. Proliferation of orbital fibroblasts and adipocytes, both in the retroocular tissue and in the perimysium of extraocular muscles, are among the most important events leading to the increased volume of orbital structures (fibroadipose tissue and extraocular muscles). The contribution of oxygen reactive species to the changes occurring in the orbit is underscored by in vitro studies. Superoxide radical stimulates orbital fibroblasts to proliferate and to produce GAG. Furthermore, hydrogen peroxide induces expression of HLA-DR and heat shock protein-72, involved in antigen recognition and T-lymphocyte recruitment. Cigarette smoking, which is probably the most important environmental factor associated with GO occurrence and maintenance, might also act, among other mechanisms, by enhancing generation of oxygen reactive species and reducing antioxidant production. Substances such as nicotinamide, allopurinol and pentoxifylline reduce superoxide- or hydrogen peroxide-induced proliferation of fibroblasts, GAG production and HLA-DR or HSP 72 expression by GO orbital fibroblasts, possibly through scavenging oxygen free radicals. Two small, non-randomized and/or uncontrolled studies investigated the effects of nicotinamide, allopurinol or pentoxifylline on GO. Favorable results were reported, but data are not fully convincing and the true effectiveness of these agents needs to be verified in randomized, controlled trials enrolling a larger number of patients. It currently seems unlikely that they may find a relevant place in the limited armamentarium available for the management of severe GO. PMID- 14757967 TI - Selenium in the treatment of autoimmune thyroiditis. AB - We recently conducted a prospective, placebo-controlled clinical study, where we could demonstrate, that a substitution of 200 microg sodium selenite for three months in patients with autoimmune thyroiditis reduced thyroid peroxidase antibody (TPO-Ab) concentrations significantly. Forty-seven patients from the initially 70 patients agreed to participate in a follow-up cross-over study for further six months. One group (n = 13), which initially received selenium continued to take 200 microg sodium selenite (Se-Se), one group stopped taking selenium (Se-0) ( n = 9), another group which received placebo started to take 200 microg selenium (n = 14) (Plac-Se) and the last group was without selenium substitution (Plac-0) (n = 11). TPO-Ab concentrations were measured at beginning and the end of the study. In the Se-Se group, the TPO-Ab concentrations further significantly p = 0.004) decreased from 625 +/- 470 U/ml to 354 +/- 321 U/ml, in the Se-0 group the TPO-Ab concentrations increased significantly p = 0.017) from 450 +/- 335 to 708 +/- 313 U/ml. In the placebo group, the TPO-Ab concentrations in those patients who were followed without selenium substitution were unchanged (1351 +/- 940 vs. 1724 +/- 1112 U/ml, p = 0.555). In contrast, the patients who received 200 microg sodium selenite after placebo, the TPO-Ab concentrations decreased significantly (p = 0.029) from 1182 +/- 723 to 643 +/- 477 U/ml. PMID- 14757968 TI - Dietary flavonoids and iodine metabolism. AB - Flavonoids have inhibiting effects on the proliferation of cancer cells, including thyroidal ones. In the treatment of thyroid cancer the uptake of iodide is essential. Flavonoids are known to interfere with iodide organification in vitro, and to cause goiter. The influence of flavonoids on iodine metabolism was studied in a human thyroid cancer cell line (FTC-133) transfected with the human sodium/iodide transporter (NIS). All flavonoids inhibited growth, and iodide uptake was decreased in most cells. NIS mRNA expression was affected during the early hours after treatment, indicating that these flavonoids can act on NIS. Pendrin mRNA expression did not change after treatment. Only myricetin increased iodide uptake. Apeginin, luteolin, kaempferol and F21388 increased the efflux of iodide, leading to a decreased retention of iodide. Instead myricetin increased the retention of iodide; this could be of use in the radioiodide treatment of thyroid cancer. PMID- 14757969 TI - Affinity of isoflavonoids for lipid bilayers evaluated with liposomal systems. AB - Three parameters, i.e., the proportion of the amount incorporated into the liposomes, the partition coefficient in a system of n-octanol/phosphate buffered saline, and the retention times by HPLC, were measured to determine the lipophilicity of isoflavonoids. The presence of a hydroxyl group at 5-position of the A-ring and a methoxyl group at 4'-position of the B-ring in the isoflavonoid structure increased the three parameters. The localization of isoflavonoids in lipid bilayers was investigated by a liposome system with fluorescent probes. The location of the isoflavonoid depended on its structure. The cytotoxicity of isoflavonoids was investigated by a colony-formation assay with Chinese hamster lung fibroblast V79 cells. The structure-activity relationship of the cytotoxic activity partly reflected those of the three parameters. This suggests that the biological activities of isoflavonoids in vitro could be attributable to their affinity for lipid components in the cases where the estrogen receptors have no role. PMID- 14757970 TI - Expression of beta-catenin in external auditory canal cholesteatoma (EACC). AB - External auditory canal cholesteatoma (EACC) is a chronic inflammation of the external auditory canal and is composed of hyperproliferative epithelium. The upward migration of the epithelial cells requires permanent breakdown and reformation of intercellular connection. This is established by the modulation of the adherent junctions consisting of an E-Cadherin-beta-catenin complex. Dissociated beta-catenin intranuclearly enables persistent activation of downstream transcription and growth factors and decreases the integrity of tissue. In our study we examined EACC and normal meatal auditory skin taken from 16 patients between 23 and 74 years of age. Immunostaining for beta-catenin was used for semiquantitative description of the specimens after assessing hematoxylin-eosin-stained slides. beta-catenin was expressed in all layers of AMS epithelium, whereas in EACC only basal layer of the matrix epithelium showed positive immunostaining for beta-catenin. In the suprabasal layer of the epithelium only faint reactivity was detectable. The immunostaining was restricted to the membrane of the cells. We assumed that either the content of membranous beta-catenin was decreased or beta-catenin was changed due to molecular modification. It is known that stimulation of endothelial cells by certain growth factors, beta-catenin is maximally phosphorylated. In regard to the increased loss of immunoreactivity for beta-catenin in the suprabasal layers of the hyperplastic EACC-matrix, we assumed bio-molecular modification or loss of beta-catenin decreasing the cell-cell-integrity. Furthermore, this might result in desquamation of keratinocytes and accumulation of dead keratin debrids. In sum, this study should be understood as a descriptive analysis of beta-catenin expression in EACC. PMID- 14757971 TI - The accumulation of alpha-Tocopherol and Retinol in the milk of water buffalo is correlated with the plasma levels of triiodothyronine. AB - Milk is the most important source of Retinol and alpha-Tocopherol for calves. These antioxidants save the food quality and prevent lipid oxidation in the mammary gland and the calf growing tissues. In Bubalus bubalis, seasonal changes for the plasma levels of both antioxidants were not found. The levels of Retinol and alpha-Tocopherol in the milk were 2 and 1.7 times higher in winter than in summer, respectively. These levels were correlated with the plasma level of triiodothyronine, and markedly increased in cows injected with triiodothyronine in summer. The cytosol from alveolar epithelial cells of mammary glands was incubated with alpha-Tocopherol and 3H-Retinol and, after gel filtration chromatography, both antioxidants were found associated with proteins migrating as a single peak of 33 kD. The amount of alpha-Tocopherol and Retinol binding proteins was 1.5 and 2.3 times higher in winter than in summer respectively. The Retinol binding proteins migrated as two bands (33 and 16 kD) by electrophoresis in denaturing and reducing conditions. Our data suggest that triiodothyronine enhances the transport of both liposoluble antioxidants through the blood-mammary barrier, and demonstrate that proteins of the mammary epithelial cells are involved in such a transport. PMID- 14757972 TI - Peroxiredoxins in antioxidant defense and redox regulation. AB - Peroxiredoxins constitute a family of peroxidases that lack prosthetic groups or catalytically active heteroatoms. Instead, their peroxidatic activity is due to a strictly conserved cysteine that is activated within a novel catalytic triad in which the cysteine thiol is coordinated to an arginine and a threonine or serine residue. Donor substrates are thiol compounds which differ between subtypes of peroxiredoxins and species. In pathogenic trypanosomatids that lack heme- or seleno-peroxidases peroxiredoxins have been shown to represent the major devices to detoxify hydroperoxides and an equivalent role may be assumed for other protozoal parasites and many bacterial pathogens. In mammals equipped with more efficient peroxidases the peroxiredoxins appear to be responsible for the redox regulation of diverse metabolic processes. The substantial differences in the cosubstrate requirements of the peroxiredoxins of pathogenic microorganisms and their mammalian host may be exploited to selectively inhibit the antioxidant defense of pathogens. Thereby, the pathogen would be more readily eliminated by the innate immune response of the host's phagocytes. PMID- 14757973 TI - Oxidative stress: the special case of diabetes. AB - The implication of oxidative stress (OS) in diabetes is a major concern for the development of therapeutics aimed at improving the metabolic and/or vascular dysfunctions of this burdening disease. Ample evidence is available suggesting that OS is present in essentially all tissues and can even be observed in prediabetic states. This raises the question of the origin of OS and suggests that, although hyperglycemia is largely linked with free radical production, its role may mainly be the aggravation of a preexisting state. Indeed other factors are also causally linked to OS, such as hormones and lipids. The main debate is about the pertinence of antioxidant therapy since the large scale clinical trials performed recently have essentially failed to show any significant improvement in metabolic or vascular disturbances of diabetic patients. However this conclusion must be tempered by the fact that they have mainly been using vitamin E +/-C; indeed many arguments suggest that either the choice or the application modalities of these substances may have been inadequate. Potential reasons for the actual failure of antioxidant therapy in diabetes are discussed; the indisputable involvement of OS in this disease still leaves hope for alternative therapeutic approaches. PMID- 14757974 TI - ADP- and oligomycin-sensitive redox behavior of F0 b thiol in ATPsynthase depends on neighbored primary structure: investigations using 14-C-labeled alpha lipoic acid. AB - Purified ATPsynthase of bovine heart mitochondria has been analyzed for its mobility and reactivity of oligomycin-sensitive sulfhydryl regions in presence of the substrate ADP and oligomycin. Labeling of thiol groups at the hydrophobic F_0 region of the ATPsynthase was increased in the enzyme initially treated with SDS, N-ethylmaleimide and dithiothreitol (modified enzyme). After dialysis or gel permeation the ATPsynthase was treated with [14C] alpha lipoic acid at a molar ratio of 35-85/1 (lipoic acid/ATPsynthase) corresponding to 4-8.6 nmol/mg protein. Under these conditions, ATPase activity of the native enzyme was significantly decreased. After preincubation with ADP, PAGE of the native, [14C] labeled enzyme revealed an increase of radioactivity at a region of 25 kDa deduced to Cys 197 of subunit b. In the modified enzyme the increase in radioactivity was found at 10 kDa. In this context, the sequence Lys-Cys-Ile around Cys 197 of subunit b suggests excessive reactivity of this thiol, as well as ready reversibility by -SH-S-S- interchange. Therefore, previously observed reaction by thiol reagents and antioxidants from outside the mitochondrion can be interpreted with Cys 197 of F0 b. It accounts for sulfhydryl unmasked by binding of ADP at F1. PMID- 14757975 TI - Hepatoprotective action of schisandrin B against carbon tetrachloride toxicity was mediated by both enhancement of mitochondrial glutathione status and induction of heat shock proteins in mice. AB - In the present study, we investigated the differential role of the mitochondrial glutathione status and induction of heat shock proteins (HSPs) 25/70 in protecting against carbon tetrachloride (CCl_4) hepatotoxicity in schisandrin B (Sch B)-pretreated mice. The time-course of Sch B-induced changes in these hepatic parameters were examined. Dimethyl diphenyl bicarboxylate (DDB), a non hepatoprotective analog of Sch B, was studied for comparison. Sch B treatment (2 mmol/kg) produced maximal enhancement in hepatic mitochondrial glutathione status as well as increases in hepatic HSP 25/70 levels at 24 h post-dosing. The stimulatory effect of Sch B then gradually subsided, but the activities of hepatic mitochondrial glutathione reductase (GR) and glutathione S-transferases (GST) as well as the level of HSP 25 remained relatively high even at 72 h post dosing. CCl_4 challenge caused significant impairment in mitochondrial glutathione status and a decrease in HSP 70 level, but the HSP 25 level was significantly elevated. While the extent of hepatoprotection afforded by Sch B pretreatment against CCl_4 was found to inversely correlate with the time elapsed after the dosing, the protective effect was associated with the ability of Sch B to maintain the mitochondrial glutathione status and/or induce further production of HSP 25 in CCl_4-intoxicated condition. On the other hand, DDB treatment (2 mmol/kg), which did not increase mitochondrial GSH level and GST activity or induce further production of HSP 25 after CCl_4 challenge, could not protect against CCl_4 toxicity. The results suggest that the enhancement of mitochondrial glutathione status and induction of HSP 25/70 may contribute independently to the hepatoprotection afforded by Sch B pretreatment. PMID- 14757976 TI - Time-dependent enhancement in mitochondrial glutathione status and ATP generation capacity by schisandrin B treatment decreases the susceptibility of rat hearts to ischemia-reperfusion injury. AB - In the present study, we examined the time-dependent changes in the mitochondrial glutathione status and ATP generation capacity in the myocardium as well as the susceptibility of the myocardium to ischemia-reperfusion (IR) injury in female Sprague Dawley rats treated with a single pharmacological dose (1.2 mmol/kg) of schisandrin B (Sch B). Sch B treatment produced a time-dependent enhancement in myocardial mitochondrial glutathione status, as evidenced by increases in myocardial mitochondrial reduced glutathione (GSH) level and activities of glutathione reductase, Se-glutathione peroxidase (GPX) and glutathione S transferases, with the response reaching maximum at 48 h post-dosing and then declining gradually to the control level at 96 h post-dosing. The enhancement of mitochondrial glutathione status was associated with an increase in myocardial ATP generation capacity, with the value peaking at 72 h post-dosing. These beneficial effects of Sch B on the myocardium was paralleled by a time-dependent decrease in the susceptibility to IR injury, with the maximum protection demonstrable at 48 h post-dosing. The cardioprotection was associated with increases in myocardial GSH level and activities of glutathione antioxidant enzymes (except for GPX whose activity was suppressed) as well as tissue ATP level/ATP generation capacity. The results suggest that Sch B treatment can precondition the myocardium by enhancing the mitochondrial glutathione status and ATP generation capacity, thereby protecting against IR injury. PMID- 14757978 TI - Molecular mechanisms of cholesterol or homocysteine effect in the development of atherosclerosis: Role of vitamin E. AB - The development of atherosclerosis is a multifactorial process in which both elevated plasma cholesterol levels and proliferation of smooth muscle cells play a central role. Numerous studies have suggested the involvement of oxidative processes in the pathogenesis of atherosclerosis and especially of oxidized low density lipoprotein. Some epidemiological studies have shown an association between high dietary intake and high serum concentrations of vitamin E and lower rates of ischemic heart disease. Cell culture studies have shown that alpha tocopherol brings about inhibition of smooth muscle cell proliferation. This takes place via inhibition of protein kinase C activity. alpha-Tocopherol also inhibits low density lipoprotein induced smooth muscle cell proliferation and protein kinase C activity. The following animal studies showed that vitamin E protects development of cholesterol induced atherosclerosis by inhibiting protein kinase C activity in smooth muscle cells in vivo. Elevated plasma levels of homocysteine have been identified as an important and independent risk factor for cerebral, coronary and peripheral atherosclerosis. However the mechanisms by which homocysteine promotes atherosclerotic plaque formation are not clearly defined. Earlier reports have been suggested that homocysteine exert its effect via H2O2 produced during its metabolism. To evaluate the contribution of homocysteine in the pathogenesis of vascular diseases, we examined whether the homocysteine effect on vascular smooth muscle cell growth is mediated by H2O2. We show that homocysteine induces DNA synthesis and proliferation of vascular smooth muscle cells in the presence of peroxide scavenging enzyme, catalase. Our data suggest that homocysteine induces smooth muscle cell growth through the activation of an H2O2 independent pathway and accelerate the progression of atherosclerosis. The results indicate a cellular mechanism for the atherogenicity of cholesterol or homocysteine and protective role of vitamin E in the development of atherosclerosis. PMID- 14757977 TI - Iron status and oxidative stress in beta-thalassemia patients in Jakarta. AB - A study on thalassemia intermedia and major patients in Jakarta was initiated to obtain a comprehensive picture of metabolic dysregulation, iron overload, oxidative stress, and cell damage. Data are presented from a group of 14 transfusion-dependent patients in an age range of 11-25 years (T) and another group of 9 frequently transfused (for at least 15 years) patients aged 17-30 years (L). A third group comprised 6 patients (aged 7 to 14 years) who had not yet obtained transfusions (N). The 21 controls (C) were voluntary students without diagnosis or clinical signs of thalassemia up to 30 years of age. The study was approved by the Ethical Clearance Board of the Medical Faculty and all blood samples from controls and patients were obtained on fully informed consent. Levels of antioxidants (vitamins A, C, E and beta-carotene) and reactive thiols are considerably decreased in transfused patients, whereas signs of iron overload and cell damage are increased (serum iron, ferritin, transferrin saturation, SGOT, SGPT, gamma-GT, bilirubin). Results can be summarized that non-transfused thalassemia intermedia patients exert slight signs of oxidative stress, and increased hemoglobin degradation but no significant indication of tissue or cell damage. This picture differs considerably from transfusion-dependent thalassemia major patients: highly significant decrease in antioxidants and thiols and tremendous iron overload and cell damage. The picture is even worsened in long term transfused patients. Iron chelation after transfusion is not sufficient in Indonesia, because it is normally (with few exceptions) applied only once together with transfusion. Hence, one major reason of the bad condition of transfusion-dependent thalassemia patients in Indonesia appears to be frequent transfusions (on the average one per month) and insufficient chelation of one treatment per month together with transfusion. PMID- 14757980 TI - Caroverine, a multifunctional drug with antioxidant functions. AB - Here we show that lipid peroxidation of liposomal membranes was suppressed in the presence of Caroverine, a spasmolytic drug used in some countries. In order to understand the mechanism of this antioxidant action of Caroverine we studied the interaction of Caroverine with superoxide radicals, hydroxyl radicals and peroxynitrite. The results of the study show that the reaction of Caroverine with O2-* radicals is of marginal significance. However, it is efficient in removing peroxynitrite and a particular high reaction constant was found for reaction with hydroxyl radicals. The strong antioxidant activity of Caroverine is therefore based both on the partial prevention of the formation and the highly active scavenging of hydroxyl radicals. PMID- 14757979 TI - Antioxidant activity in vitro of two aromatic compounds from Caesalpinia sappan L. AB - Two antioxidant compounds were isolated from C. sappan L by multiple steps of column chromatography and thin layer chromatography in succession with superoxide scavenging assay as activity monitor. Structures of the two compounds were determined by spectroscopic methods as 1',4'-dihydro-spiro[benzofuran-3(2H),3' [3H-2]benzopyran]-1',6',6',7'-tetrol (compound 1) and 3-[[4,5-dihydroxy 2(hydroxymethyl) phenyl]-methyl]-2,3-dihydro-3,6-benzofurandiol (compound 2). Characterization of antioxidant properties of these two compounds was done by determining the inhibitory effect on xanthine oxidase activity as well as scavenging effect on superoxide anion and hydroxyl radicals. Our results indicated that compounds 1 and 2 inhibited xanthine oxidase activity and scavenged superoxide anion and hydroxyl radicals. Compounds 1 and 2 possessed similar radical scavenging activities as ascorbic acid, and they were more effective than other well-known antioxidants such as alpha-tocopherol, beta carotene, and BHT. As inhibitors of free radical formation, compounds 1 and 2 were more effective than all the other antioxidants tested. In conclusion, compounds 1 and 2 can be regarded as primary antioxidants with radical-scavenging and chain-breaking activities as well as secondary antioxidants with inhibitory effect on radical generation. PMID- 14757981 TI - Isolated erythrocyte membranes of transfusion-dependent and non-transfused thalassemia patients in Jakarta, investigated by electron paramagnetic resonance spectroscopy. AB - Erythrocyte membrane structural parameters were studied in transfusion-dependent beta-thalassemia patients, in long-term transfused patients (regularly transfused < 15 years), and in those who had not yet obtained transfusions. Controls were voluntary students up to 30 years of age without diagnosis or clinical signs of thalassemia. Membranes were isolated and investigated by sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE) and electron paramagnetic resonance (EPR) spectroscopy. Data obtained from the thiol-reactive spin label N-ethyl maleimidoproxyl reveal immobilization of protein environment in erythrocyte membranes from thalassemic patients. SDS-PAGE shows both degradation and aggregation of membrane proteins. Thalassemic erythrocyte membranes exert higher order parameters in the hydrophobic region as determined by 16-doxyl-stearic acid. Rotational correlation times of this spin label increase only in transfused patients. Polarity is higher in membranes of all patients than in controls. In the polar interface, order parameters obtained from 5-doxyl-stearic acid increase in non-transfused and decrease in transfusion-dependent patients as compared with controls. Transfused patients exert increasing membrane order in the hydrophobic region and counter-currently decreasing order in the polar interface indicating loss of membrane integrity along with the loss of fluidity and polarity gradients and the loss the energetic barrier function of the membrane. PMID- 14757982 TI - The association between negative and dysexecutive syndromes in schizophrenia: a cross-cultural study. AB - This paper examined the relationship between the 'negative syndrome' (NS) and the neuropsychological 'dysexecutive syndrome' (DES) in schizophrenia. The study also examined whether any relationship that exists between the NS and the DES holds equally for British and Japanese subjects. We compared 26 Japanese with 17 British schizophrenic patients, divided into 'mild' and 'severe' NS groups, on the basis of performance on neuropsychological tests, including the 'Behavioural Assessment of Dysexecutive Syndrome' (BADS). We found that patients with severe NS showed more everyday executive deficits than those with mild NS. The severity of NS was correlated with executive competence. The association between NS and the BADS performance was closer than that between NS and other conventional executive measures. These findings were not influenced by cultural differences between Japanese and British subjects, and, hence, suggested the existence of culture-neutral neurobehavioural processes. PMID- 14757983 TI - Subtyping obsessive-compulsive disorder: neuropsychological correlates. AB - We administered neuropsychological measures considered sensitive to prefrontal dysfunction (both orbitofrontal and dorsolateral prefrontal neocortex) to obsessive-compulsive disorder (OCD) patients and control subjects. OCD subjects exhibited performance deficits, in comparison to community controls, on three measures sensitive to orbitofrontal neocortex dysfunction. Contrary to expectation, OCD patients also exhibited performance deficits on measures sensitive to dorsolateral prefrontal neocortex dysfunction. However, distinct neurocognitive profiles emerged when we examined the impact of comorbid schizotypal personality features on neuropsychological test performance. Primary OCD patients displayed impaired performance on measures sensitive to orbitofrontal dysfunction; however, they did not differ from control subjects on tests of dorsolateral function. OCD subjects presenting with schizotypal personality features performed poorly not only on tests sensitive to orbitofrontal dysfunction, but also on tests sensitive to dorsolateral dysfunction. Findings suggest that OCD can be subdivided into clinical subtypes, and distinct prefrontal subsystems may be differentially involved in these subtypes. PMID- 14757984 TI - Narcolepsy: pathophysiology and neuropsychological changes. AB - Narcolepsy is now recognized as a distinctive disorder with specific pathophysiology and neurochemical abnormalities. Findings on the role of the neuropeptide hypocretin are opening new avenues of research and new strategies for therapy. Recently, neuropsychological and electrophysiological studies have provided evidence for reduced memory performance on standard memory tests in addition to subjective complaints of forgetfulness which may be related to changes in attentional processing. Further studies are, however, necessary to clarify the neuropsychological profile in narcolepsy. This review focuses on the recent advances in understanding narcolepsy. PMID- 14757985 TI - Involuntary vocalisations and a complex hyperkinetic movement disorder following left side thalamic haemorrhage. AB - A variety of involuntary speech phenomena as for example palilalia have been described as consequences of neurological disorders. Palilalia is the involuntary repetition of syllabels, words and phrases in ongoing speech. We describe a 73 year old woman who suffered from a hypertensive thalamic haemorrhage. MRI revealed that the lesion was predominantly located within the pulvinar, extending to the lateroposterior thalamic nuclei and to the pretectal area with possible involvement of the medial geniculate body. Few months after the event she developed involuntary vocalisations with whole words and meaningless syllables being rapidly reiterated. In contrast to typical palilalia these vocalisations were not meaningfully related to the ongoing speech of the patient. In addition, the patient developed a complex hyperkinetic movement disorder with right-sided painful hemidystonia and bilateral clonic jerks and a right-sided postural tremor. PMID- 14757986 TI - Cognitive and emotional dysfunction after central pontine myelinolysis. AB - The case of a 67-year-old right-handed Chinese man with Central Pontine Myelinolysis [CPM] is described to illustrate the resulting cognitive and emotional disturbances. A comparison of the data in this report with that in published studies suggests that ethnicity does not seem to have much effect on the symptoms of CPM. Possible underlying neural-pathological mechanisms are discussed. This case further substantiates the speculation that the brainstem plays a role in higher cognitive processes and emotional regulation. PMID- 14757987 TI - Developmental prosopagnosia: a review. AB - This article reviews the published literature on developmental prosopagnosia, a condition in which the ability to recognize other persons by facial information alone has never been acquired. Due to the very low incidence of this syndrome, case reports are sparse. We review the available data and suggest assessment strategies for patients suffering from developmental prosopagnosia. It is suggested that developmental prosopagnosia is not a unitary condition but rather consists of different subforms that can be dissociated on the grounds of functional impairments. On the basis of the available evidence, hypotheses about the aetiology of developmental prosopagnosia as well as about the selectivity of deficits related to face recognition are discussed. PMID- 14757988 TI - Establishing mathematical laws of genomic variation. AB - As the biological arm of the Rasch community, genomic measurement is concerned with asserting and testing hypotheses regarding the quantitative status of genomic variables, including alleles, genotypes, gene expression levels, and phenotypes, as well as DNA, RNA, and protein sequence information. The defining goal of this scientific paradigm, in contrast to the sample-dependent model fitting and deterministic hypothesis testing of classical statistical genetics, is the identification, validation, and maintenance of a common unit of genomic measurement that maintains its magnitude and meaning, within an allowable range of error, regardless of the laboratory technology used to generate outcomes or the particular group of individuals or organisms under investigation. Such an invariant metric, the basis of a standard genometric scale and associated system of genomic metrology, can be identified, validated, and maintained through 1) routine implementation of the Rasch family of measurement models to construct sample- and scale-free measures from different types of genomic data and 2) cross calibration of genomic measurement instruments between and among researchers, laboratories, universities, corporations, and databases. This manuscript provides an introductory overview of the guiding principles of fundamental measurement theory and the work of Rasch, connects these concepts to well-known tenets of population genetics, and highlights the potential benefits, both theoretical and applied, associated with achieving objectivity in genomic measurement. PMID- 14757989 TI - Comparing traditional and Rasch analyses of the Mississippi PTSD Scale: revealing limitations of reverse-scored items. AB - This study examined whether Rasch analysis could provide more information than true score theory (TST) in determining the usefulness of reverse-scored items in the Mississippi Scale for Posttraumatic Stress Disorder (M-PTSD). Subjects were 803 individuals in inpatient PTSD units at 10 VA sites. TST indicated that the M PTSD performed well and could be improved slightly by deleting one item. Factor analysis using raw scores indicated that the reverse-scored items formed the second factor and had poor relationships with normally scored items. However, since item-total correlations supported their usefulness, they were kept. The subsequent Rasch analysis indicated that five of the seven worst fitting items were reverse-scored items. We concluded that using reversed items with disturbed patients can cause confusion that reduces reliability. Deleting them improved validity without loss of reliability. The study supports the use of Rasch analysis over TST in health research since it indicated ways to reduce respondent burden while maintaining reliability and improving validity. PMID- 14757990 TI - Evaluating judge performance in sport. AB - Many sports, such as, gymnastics, diving, ski jumping, and figure skating, use judges' scores to determine the winner of a competition. These judges use some type of rating scale when judging performances (e.g., figure skating: 0.0 - 6.0). Sport governing bodies have the responsibility of setting and enforcing quality control parameters for judge performance. Given the judging scandals in figure skating at the 1998 and 2002 Olympics, judge performance in sport is receiving greater scrutiny. The purpose of this article is to illustrate how results from Rasch analyses can be used to provide in-depth feedback to judges about their scoring patterns. Nine judges' scores for 20 pairs of figure skaters who competed at the 2002 Winter Olympics were analyzed using a four-faceted (skater pair ability, skating aspect difficulty, program difficulty, and judge severity) Rasch rating scale model that was not common to all judges. Fit statistics, the logical ordering of skating aspects, skating programs, and separation indices all indicated a good fit of the data to the model. The type of feedback that can be given to judges about their scoring pattern was illustrated for one judge (USA) whose performance was flagged as being unpredictable. Feedback included a detailed description of how the rating scale was used; for example, 10% of all marks given by the American judge were unexpected by the model (Z > |2|). Three figures illustrated differences between the judge's observed and expected marks arranged according to the pairs' skating order and final placement in the competition. Scores which may represent "nationalistic bias" or a skating order influence were flagged by looking at these figures. If sport governing bodies wish to improve the performance of their judges, they need to employ methods that monitor the internal consistency of each judge as a many-facet Rasch analysis does. PMID- 14757991 TI - The effect of sample size for estimating Rasch/IRT parameters with dichotomous items. AB - Thirteen samples were randomly drawn from the normative database for the latest edition of Knox's Cube Test-Revised (KCT-R). Parameter estimates for the Rasch model and two and three parameter logistic models were derived and compared. Sample size influenced these estimates as might be expected. Rasch parameter estimates consistently showed the smallest values by sample size using a goodness of fit index. PMID- 14757992 TI - Equating student satisfaction measures. AB - Colleges and universities conduct student satisfaction studies for many important policy making reasons. However the differences in instrumentation and the use of students' self-reported ratings of satisfaction makes such decisions sample-, instrument-, and institution-dependent. A common metric of student satisfaction would assist decision makers by providing a richness of information not typically obtained. The present study investigated the extent to which two nationally known instruments of student satisfaction could be scaled on the same quantitative metric. Pseudo-common item equating (Fisher, 1997) based on five link items of low and high endorsability enabled comparisons of "similar, but not identical items, from different instruments, calibrated on different samples" (p. 87). Results suggest that both instruments measured similar constructs and could be reasonably used to create a single, common metric. While samples used in the experiment were less than ideal, results clearly demonstrated the usefulness and reasonability of the pseudo-common item equating process. PMID- 14757994 TI - Rasch model estimation: further topics. AB - Building on Wright and Masters (1982), several Rasch estimation methods are briefly described, including Marginal Maximum Likelihood Estimation (MMLE) and minimum chi-square methods. General attributes of Rasch estimation algorithms are discussed, including the handling of missing data, precision and accuracy, estimate consistency, bias and symmetry. Reasons for, and the implications of, measure misestimation are explained, including the effect of loose convergence criteria, and failure of Newton-Raphson iteration to converge. Alternative parameterizations of rating scales broaden the scope of Rasch measurement methodology. PMID- 14757993 TI - Treating test-item nonresponse. AB - This study presents the results of an empirical investigation into the effects of nonresponse on the measurement of student ability in large scale educational achievement studies. The analyses explored the bias in student and national ability estimate distributions relating to particular psychometric treatments of nonresponse. A range of replicated analyses were undertaken using data collected from a crossnational study of reading achievement. The relative merits of these treatments are summarized in conclusion, and implications for normative and methodological research and practice are considered. It is suggested that environmental and psychological antecedents of nonresponse need to be determined and that related variables be included as essential components in item response modelling. PMID- 14757995 TI - Use of the glycemic index: effects on feeding patterns and exercise performance. AB - The focus of this paper is on the glycemic index (GI) that provides effectual information on planning nutritional strategies for carbohydrate (CHO) supplementation in exercise. Related research has suggested that the GI can be used as a reference guide for the selection of an ideal CHO supplement in sports nutrition. Recently, the manipulation of GI of CHO supplementation in optimizing athletic performance has provided an exciting new research area in sports nutrition. There is a growing evidence to support the use of the GI in planning the nutritional strategies for CHO supplementation in sports. The optimum CHO availability for exercise has been demonstrated by manipulating the GI of CHO. Research has shown that a low GI CHO-rich meal is a suitable CHO source before prolonged exercise in order to promote the availability of the sustained CHO. In contrast, a high GI CHO-rich meal appears to be beneficial for glycogen storage after the exercise by promoting greater glucose and insulin responses. The prescribed feeding patterns of CHO intake during recovery and prior to exercise on glycogen re-synthesis and exercise metabolism have been studied in the literature. However, the studies on the subject are still limited, leaving some open questions waiting for further empirical evidences. The most significant question is whether CHO supplementation before and after exercise is beneficial when consumed as large feedings or as a series of snacks. Further research is needed on the effect of feeding patterns on exercise performance. PMID- 14757996 TI - Kinetics of oxygenation in inactive forearm muscle during ramp leg cycling. AB - This study was carried out to determine whether hemodynamics in inactive forearm muscle during ramp leg cycling is affected from the ventilatory threshold (VT) and respiratory compensation point (RCP), at which the rate of increase in ventilation (VE) against power output begins to increase abruptly. Change in hemodynamics was evaluated by change in oxygenation index (difference between concentrations of oxygenated hemoglobin and deoxygenated hemoglobin, HbD) measured using near-infrared spectrometry (NIRS). Each subject (n=9) performed 4 min constant-work-rate leg cycling and subsequent ramp leg cycling at an increasing rate of 10 watts.min(-1) in power output. The work rates at VT, RCP and peak oxygen uptake (VO(2 peak)) were 107 +/- 11, 172 +/- 21 and 206 +/- 20 watts, respectively. The rates of increase in VE between 10-watt leg cycling, VT, RCP and VO(2 peak) were 0.19 +/- 0.03, 0.44 +/- 0.07 and 1.32 +/- 0.47 l.min( 1).watts(-1), respectively. In one subject, HbD started to decrease during ramp exercise from the VT, and the rate of decrease increased at a high intensity of exercise. In eight subjects, although no decrease in HbD from the VT was observed, HbD showed a sudden drop at a high intensity of exercise. The work rate at which HbD began to decrease at a high intensity of exercise was 174 +/- 23 watts. This work rate was not significantly different from that at the RCP and was significantly correlated with that at the RCP (r=0.72, P<0.05). The results suggest that the abrupt increase in VE from the RCP affects hemodynamics, resulting in a decrease in HbD in inactive forearm muscle. PMID- 14757997 TI - Calorie restricted diet and urinary pentosidine in patients with rheumatoid arthritis. AB - Low-energy diets and fasting have suppressive effects on rheumatoid arthritis. It was reported recently that urine levels of pentosidine (i.e., an advanced glycation end product formed by glycosylation) is associated with the activity of rheumatoid arthritis. We conducted a regimen of caloric restriction combined with fasting in patients with rheumatoid arthritis, and then evaluated urinary pentosidine levels. Ten patients with rheumatoid arthritis underwent a 54-day caloric restriction program. Urinary pentosidine levels were measured and the Lansbury Index were determined by examining the clinical features, blood biochemistry and the inflammation activity of rheumatoid arthritis on days 0, 25 and 54. On day 0, the mean urinary pentosidine level of patients with rheumatoid arthritis was significantly higher than that of the control subjects. On day 54, the mean body weight had reduced due to caloric restriction. The mean values of the erythrocyte sedimentation rate and the Lansbury Index of patients both significantly decreased during the study. In addition, although the urinary pentosidine levels showed no significant difference between day 0 and 25, it was significantly decreased at the end of the study (day 54). The study showed that under a low energy diet a reduction of disease activity in rheumatoid arthritis was accompanied with a reduction of the urinary pentosidine. PMID- 14757998 TI - Validity of the body mass index and fat mass index as an indicator of obesity in children aged 3-5 year. AB - The validity of the BMI and fat mass index (FMI) as indicators of obesity was evaluated in a group of 3-5 yr old (n=486) children. Bioelectrical impedance analysis (BIA) was measured (using 50 kHz and tetrapolar electrodes) in order to calculate percent fat mass (%FM) and FMI (fat mass/stature squared). For boys, obesity was defined as > or =20%FM. For girls, the cutoff for obesity was > or =25%FM. However, obesity was defined as a BMI at or above the 90th percentile of age- and sex-specific data in this study. The percentile cutoffs for FMI were the same as for BMI using the same sample. There were correlations between BMI or FMI and %FM, but there was no significant correlation between BMI or FMI and stature. Therefore, it appears that both the BMI and FMI in this study are far more useful indices with which to assess obesity, and are reasonable indicators of fatness. However, with the use of %FM by BIA as the criterion for obesity, BMI and FMI had high specificities (95.5-96.4% for BMI and 99.5-100% for FMI) and lower but variable sensitivities (30.4-37.5% for BMI and 42.9-68.8% for FMI). Thus, almost all children who were not obese were classified correctly. In contrast, many obese children were not correctly identified by BMI and FMI. Therefore, we conclude that BMI should be used with caution as an indicator of childhood obesity. The new recommendations based on the FMI approach for defining childhood obesity are associated with a level of sensitivity that is somewhat higher than that of the BMI approach. Caution should, however, be used in generalizing from the findings in this study, and a further investigation of the issue is required. PMID- 14757999 TI - Investigation of the release test method for the topical application of pharmaceutical preparations: release test of cataplasm including nonsteroidal anti-inflammatory drugs using artificial sweat. AB - A simple procedure for determining the in vitro release profile of a cataplasm for use in a quality control procedure has been developed. Since the disk assembly in the USP for patch dosage forms was unsuited for use in a release test due to penetration of the dissolution medium into the cataplasm from the screw part of the device and the cataplasm swelled, new holders were designed. In the new holder, a cataplasm is held in position by sandwiching it between a stainless steel O-ring and a silicon O-ring on the stainless steel board, 2 acrylic boards hold the O-rings and the stainless steal board, and the entire assembly is placed at the bottom of the dissolution vessel. The release profile was determined using the "Paddle over Disk" method in USP26. Furthermore, in order to prevent the swelling of the cataplasm, artificial sweat was used as the dissolution medium. The release profiles of the nine marketed brands of cataplasm containing indomethacin, ketoprofen, and flurbiprofen, respectively, were determined over a 12-h period. By adjusting the ion concentration and volume of the media, and the release surface-area of the cataplasm exposed to each medium, the procedure was found to be reproducible for in vitro release characterization of nine marketed brands. This shows that this technique can be used as a quality control tool for assuring product uniformity. PMID- 14758000 TI - The combination effect of L-arginine and NaCl on bitterness suppression of amino acid solutions. AB - The purpose of the present study was to quantify the degree of suppression of the bitterness of two amino acids (L-isoleucine (L-Ile), and L-phenylalanine (L-Phe)) which could be achieved by the addition of various test chemicals, and to examine the mechanism of this bitterness suppression. The test chemicals used were two sweeteners (sucrose, aspartame), NaCl, various acidic (L-aspartic acid, L glutamic acid), or basic (L-histidine, L-lysine and L-arginine) amino acids, tannic acid and phosphatidic acid. The combination of L-arginine (L-Arg) and NaCl together was the most effective in reducing the bitterness of 100 mM L-Ile and L Phe solutions in human gustatory sensation tests. Even in bitterness of 0.1 mM quinine solution, L-Arg was also successful in reducing the bitterness. This bitterness-suppression effect was specific to L-Arg and not to the other basic amino acids. No comparable taste-masking effect was observed for the acidic amino acids. The artificial taste sensor failed to predict completely the bitterness suppressing effect of L-Arg. It seems likely that the bitterness-suppressing effect of L-Arg is mediated not only by binding at the receptor site, but also elsewhere in the process of bitterness perception, such as a direct effect on the sodium channel. It is conjectured that the guanidinium group of L-Arg may interact with sodium channels in taste bud membranes. PMID- 14758001 TI - Synthesis, anticancer and antibacterial activity of some novel mononuclear Ru(II) complexes. AB - In search of potential anticancer drug candidates in ruthenium complexes, a series of mononuclear ruthenium complexes of the type [Ru(phen)(2)(nmit)]Cl(2) (Ru1), [Ru(bpy)(2)(nmit)]Cl(2) (Ru2), [Ru(phen)(2)(icpl)]Cl(2) (Ru3), Ru(bpy)(2)(icpl)]Cl(2) (Ru4) (phen=1,10-phenanthroline; bpy=2,2'-bipyridine; nmit=N-methyl-isatin-3-thiosemicarbazone, icpl=isatin-3-(4-Cl phenyl)thiosemicarbazone) and [Ru(phen)(2)(aze)]Cl(2) (Ru5), [Ru(bpy)(2)(aze)]Cl(2) (Ru6) (aze=acetazolamide) and [Ru(phen)(2)(R tsc)](ClO(4))(2) (R=methyl (Ru7), ethyl (Ru8), cyclohexyl (Ru9), 4-Cl-phenyl (10), 4-Br-phenyl (Ru11), and 4-EtO-phenyl (Ru12), tsc=thiosemicarbazone) were prepared and characterized by elemental analysis, FTIR, (1)H-NMR and FAB-MS. Effect of these complexes on the growth of a transplantable murine tumor cell line (Ehrlich Ascites Carcinoma) and their antibacterial activity were studied. In cancer study the effect of hematological profile of the tumor hosts have also been studied. In the cancer study, the complexes Ru1-Ru4, Ru10 and Ru11 have remarkably decreased the tumor volume and viable ascitic cell count as indicated by trypan blue dye exclusion test (p<0.05). Treatment with the ruthenium complexes prolonged the lifespan of Ehrlich Ascites Carcinoma (EAC) bearing mice. Tumor inhibition by the ruthenium chelates was followed by improvements in hemoglobin, RBC and WBC values. All the complexes showed antibacterial activity, except Ru5 and Ru6. Thus, the results suggest that these ruthenium complexes have significant antitumor property and antibacterial activity. The results also reflect that the drug does not adversely affect the hematological profiles as compared to that of cisplatin on the host. PMID- 14758002 TI - The reaction rate of edaravone (3-methyl-1-phenyl-2-pyrazolin-5-one (MCI-186)) with hydroxyl radical. AB - The pyrazoline derivative edaravone is a potent hydroxyl radical scavenger that has been approved for attenuation of brain damage caused by ischemia-reperfusion. In the present work, we first determined the rate constant, k(r), at which edaravone scavenges radicals generated by a Fenton reaction in aqueous solution in the presence of the spin trap agent, 5,5-dimethyl-1-pyrroline-N-oxide (DMPO), which competed with edaravone. We detected the edaravone radicals in the process of hydroxyl radical scavenging and found that edaravone reacts with hydroxyl radical around the diffusion limit (k(r)=3.0 x 10(10) M(-1) s(-1)). The EPR (electron paramagnetic resonance) spectrum of the edaravone radical was observed by oxidation with a horseradish peroxidase-hydrogen peroxide system using the fast-flow method. This radical species is unstable and changed to another radical species with time. In addition, it was found that edaravone consumed molecular oxygen when it was oxidized by horseradish peroxidase (HRP)-H(2)O(2) system, and that edaravone was capable of providing two electrons to the electrophiles. The possible mechanisms for oxidation of edaravone were investigated from these findings. PMID- 14758003 TI - Synthesis, structure and properties of N-acetylated derivatives of methyl 5-amino 1H-[1,2,4]triazole-3-carboxylate. AB - Methyl 5-amino-1H-[1,2,4]triazole-3-carboxylate hydrochloride (1). and free ester (2). were obtained and 2 was reacted with Ac(2)O to give the acetylated products 3-6. Compounds 1-6 were studied using HPLC, GC-MS, FTIR and multinuclear NMR spectroscopy, including the cross-polarisation magic angle spinning (CPMAS) technique. The results of the acetylation of 2 were compared to those of the acetylation of 5-amino-1H-[1,2,4]triazole, and for 2 a significant decrease in the susceptibility to acetylation was found. The reaction of 2 with Ac(2)O at 20 degrees C, regardless of the amount and the concentration of the latter, including neat Ac(2)O, proceeds fully regioselectively and leads to one product: methyl 1-acetyl-5-amino-1H-[1,2,4]triazole-3-carboxylate (3). In sharp contrast to 5-amino-1H-[1,2,4]triazole, neither an additional monoacetylated isomer, whether annular or exocyclic, nor any diacetylated derivative could be detected. The diacetylation of 2 requires the process to be carried out in neat boiling Ac(2)O and, as in the case of 5-amino-1H-[1,2,4]triazole, gives two diacetylated isomers. These are methyl 1-acetyl-3-(acetylamino)-1H-[1,2,4]triazole-5 carboxylate (4) and 1-acetyl-5-(acetylamino)-1H-[1,2,4]triazole-3-carboxylate (5). Hypothetical pathways of their formation have been suggested. A mixture of 4 and 5 upon hydrolysis of the ring acetyl group gives the monoacetylated derivative methyl 5-(acetylamino)-1H-[1,2,4]triazole-3-carboxylate (6). The spectroscopic, structural and conformational characteristics of compounds 1-6 have been given and methods for their preparation have been provided. PMID- 14758004 TI - Protection of protein secondary structure by saccharides of different molecular weights during freeze-drying. AB - The protective effects of saccharides with various molecular weights (glucose, maltose, maltotriose, maltotetraose, maltopentaose, maltoheptaose, dextran 1060, dextran 4900, and dextran 10200) against lyophilization-induced structural perturbation of model proteins (BSA, ovalbumin) were studied. Fourier transform infrared (FT-IR) analysis of the proteins in initial solutions and freeze-dried solids indicated that maltose conferred the greatest protection against secondary structure change. The structure-stabilizing effect of maltooligosaccharides decreased in increasing the number of saccharide units. Larger molecules of dextran also showed a smaller structure-stabilizing effect. Increasing the effective saccharide molecular size by a borate-saccharide complexation reduced the protein structure-stabilizing effect of all of the saccharides except glucose. The results indicate that the larger saccharide molecules, and/or the complex formation with borate ion, reduce the free and accessible hydroxyl groups to interact with and stabilize the protein structure by a water-substitution mechanism. PMID- 14758006 TI - A synthesis of heteroaromatic analogues of 1-methyl-1,2,3,4 tetrahydroisoquinoline using the Pummerer-type cyclization reaction: observation of tandem cyclization reaction. AB - The sulfoxides 7b and 7d carrying thiophene or benzothiophene as heteroaromatic nucleophiles, when treated with trifluoroacetic anhydride at room temperature (Pummerer reaction), underwent an intramolecular alkylation in an exclusive manner to yield 4,5,6,7-tetrahydro-7-methyl-4-phenylsulfanylthieno[2,3-c]pyridine 6-carbaldehyde (10) and the corresponding benzothiophene derivative (12b) in high yields, respectively. Thus, this route provides biologically interesting nitrogen heterocycles (1b) and (2b). On the other hand, the sulfoxide (7c) carrying benzofuran as a nucleophile on reaction with TFAA yielded not only the Pummerer type cyclization product (12a), but also the diastereoisomeric tandem cyclization products (13) and (14) having a noble 11-aza-2-oxa-7 thiatricyclo[4.3.3.0(1,5)]dodecane ring system (B). The formation of these products can be readily rationalized by the intervention of the oxonium ion intermediate (21). PMID- 14758005 TI - Diastereomeric selective effects for growth inhibition of synthesized mini parallel double-stranded peptides on Escherichia coli and Staphylococcus aureus. AB - The synthesis of a series of mini double-stranded peptides containing chiral-x Phe-y-Phe-peptide residues and the diastereomeric selective effects of these compounds on Escherichia coli NIHJ JC-2 and Staphylococcus aureus FDA 209P growth are described. In the case of bis(y-Phe-x-Phe)-N,N-ethane-1,2-diamine, bis(y-Phe x-Phe)-N,N-buthane-1,4-diamine, bis(y-Phe-x-Phe)-N,N-hexane-1,6-diamine, and bis(y-Phe-x-Phe)-N,N-dodecane-1,12-diamine, etc., the four configurations, (L-, L ), (D-, L-), (L-, D-) and (D-, D-), where the symbols x- and y- represent optical isomers with L- and D- forms, were used to investigate the relationship between chirality and antibacterial activity. The level of activity increased in the following order: (L-, L-)<(D-, D-)<(L-, D-)<(D-, L-). The data show that (D-, L-) chilarity is more potent than (L-, L-) chilarity. Then, these results suggest that the -y-Phe-x-Phe Phe-sequence in the double-stranded peptide has anti bacterial activity and the chirality of -y-Phe-x-Phe affects the anti-bacterial activity. Our results show that the uptake by penetration through the membrane of bis(y-Phe-x-Phe)(2)-Spacers is a first step in the expression of anti-bacterial activity. This study provides new insights in the chirality-antibacterial activity relationships of a series of mini double-stranded peptides. PMID- 14758007 TI - The interaction of monosubstituted benzenes with the stationary liquid in gas liquid chromatography. AB - In gas liquid chromatography (GLC), the relative retention values log gamma was mainly expressed by van der Waals energy (the sum of the dispersion E(dis) and repulsive E(rep) energies) to the interactions between monosubstituted benzene derivatives and the nonpolar stationary liquid as squalane. The single exception was that of anilines, and it was corrected by the electrostatic energy (E(ES)) due to C-H/pi hydrogen bond. When the stationary liquid changed from the nonpolar to polar, log gamma was estimated by the inductive interaction energy (included in E(ES)) in addition to the sum of E(dis) and E(rep). In the benzene solution, the relative equilibrium values log K/K(o) introduced from the interactions between phenol and substituted benzene derivatives were estimated by E(ES). The E(ES) of COCH(3), CO(2)C(2)H(5) groups is especially originated in the excited dipole moments micro(e). The relative frequency values log nu/nu(o) derived from O-H or O-D stretching vibration of phenol or methanol-D gave the correlation to E(ES) as well as log K/K(o). That of anilines-methanol-D however had been out of a linear relation to E(ES). The cause is concluded that the aniline-methanol-D is making the proton transfer structure from the discussion about the proton affinity (PA) of the base. PMID- 14758008 TI - Complete assignment of bilberry (Vaccinium myrtillus L.) anthocyanins separated by capillary zone electrophoresis. AB - Capillary zone electrophoresis (CZE) mobilities of fifteen anthocyanins in bilberry extract were completely characterized. Four minor anthocyanins in bilberry extract (malvidin 3-O-alpha-L-arabinopyranoside (Mv 3-ara), peonidin 3-O beta-D-galactopyranoside (Pn 3-gal), peonidin 3-O-alpha-L-arabinopyranoside (Pn 3 ara), and petunidin 3-O-alpha-L-arabinopyranoside (Pt 3-ara)) that remained unidentified in our previous CZE study were isolated from the bilberry extract, and the chemical structures were assigned by NMR and MS. Their CZE mobilities were then precisely examined together with those of other major anthocyanins in the extract. When the CZE mobilities of the fifteen anthocyanins assigned here were plotted against their molecular weight/numbers of free phenolic group, it was found that separation of anthocyanins by CZE is primarily determined by the type of conjugated sugar present, and secondly by the aglycon structure. PMID- 14758009 TI - New triterpenoid saponins from the roots of Sinocrassula asclepiadea. AB - Five new triterpenoid monodesmosides (sinocrassulosides I-V, 1-5) and six bisdesmosides (sinocrassulosides VI-XI, 6-11), in which 2-11 possess different acyl groups in the glycosidic moieties, were isolated from the roots of Sinocrassula asclepiadea FRANCH. Sinocrassulosides VI (4) and V (5) also contained a novel A-seco aglycone in their structures. All of the structures were determined on the basis of spectroscopic and physicochemical evidence. PMID- 14758010 TI - Preparation on oligostilbenes of isorhapontigenin by oxidative coupling reaction. AB - Four new compounds 1-4 were obtained from an oxidative coupling reaction of (E) isorhapontigenin using FeCl(3) as oxidant. Their structures and stereochemistry were determined on the basis of spectroscopic evidence [UV, IR, MS, (1)H-, (13)C NMR, NOE and 2D NMR], and their possible formation mechanisms were also discussed, respectively. PMID- 14758011 TI - Application of nilvadipine solid dispersion to tablet formulation and manufacturing using crospovidone and methylcellulose as dispersion carriers. AB - Nilvadipine (NIL) solid dispersion using crospovidone (Cross-linked-N-vinyl-2 pyrolidone, cl-PVP) and methylcellulose (MC) as carriers was applied to tablet formulation. Several grades of cl-PVP and MC were used, and their influence on tablet properties such as hardness, disintegration, dissolution and chemical stability were investigated. The agitation granulation method was used for preparation of solid dispersion granules, and the granules were compressed using a rotary tableting machine, and finally the obtained tablets were coated with film. As the particle size of cl-PVP decreased, hardness and apparent solubility were increased, while dissolution rate was lowered. When a higher viscosity grade of MC was used, hardness and dissolution rate were increased, and apparent solubility did not change. All batches of tablets were chemically stable at 40 degrees C, 75% relative humidity (R.H.) for six months. Finally, tablets with enhanced dissolution properties were obtained by using Polyplasdone XL-10 and Metolose SM-25 as the grades of cl-PVP and MC, respectively. These formulation tablets showed higher solubility and dissolution rate during storage as well as initial indicating good physical stability. PMID- 14758012 TI - Reaction of beta-ethoxyvinyl lithiums generated from phenyltellanyl- and ethyltellanylacetaldehyde diethyl acetals with aldehydes and ketones and successive hydrations. AB - Reactions of alpha-tellanyl-beta-ethoxyvinyl lithiums of aldehydes and ketones proceeded in good to high yields and the successive treatment with acids gave the alpha-tellanyl alpha,beta-unsaturated aldehydes. alpha-Tellanyl alpha,beta unsaturated aldehydes easily transformed to more useful compounds. PMID- 14758013 TI - Synthesis of 1-benzothiepine and 1-benzazepine derivatives as orally active CCR5 antagonists. AB - Quaternary ammonium benzocycloheptene compound 1 has previously been reported as a clinical candidate for an injectable CCR5 antagonist. In order to develop an orally active CCR5 antagonist, derivatives of tertiary amine benzocycloheptene 2, the chemical precursor to 1, were investigated. The benzocycloheptene ring was converted to benzothiepine and benzazepine rings and it was found that these changes could enhance the potency of tertiary amine derivatives. In particular, the 1-benzothiepine-1,1-dioxide 11b and the N-methyl-1-benzazepine 18 showed increased activity and good preliminary pharmacokinetic properties. The synthesis of 1-benzothiepine and 1-benzazepine derivatives and their activity are described. PMID- 14758014 TI - A modified method using static head-space gas chromatography for determining the stability constants of 1-alkanol/alpha-cyclodextrin complexation. AB - A modification of the conventional static head-space gas chromatography method (SHSGC method) to determine stability constants for 1-alkanol/alpha-CD inclusion complexes was investigated. The 1 : 1 stability constants determined by this modified SHSGC method are in reasonable agreement with the corresponding values reported previously. The modified SHSGC method precludes the necessity of the calibration curve by the use of Henry's law constant of guest. Consequently, the modified SHSGC method is more advantageous than the conventional SHSGC method because the experimental time required for determination of the stability constant is markedly reduced. PMID- 14758015 TI - Synthesis of 2-N,N-dimethylaminomethyl-2,3,3a,12b-tetrahydrodibenzo-[b,f]furo[2,3 d]oxepin derivatives as potential anxiolytic agents. AB - New synthesis approaches that have led to a series of novel tetrahydrodibenzo[b,f]furo[2,3-d]oxepin derivatives are described. According to preliminary data these novel tetracycles can be useful intermediates for the preparation of potential new therapeutic agents. PMID- 14758016 TI - Catalytic activity for decomposition of hydrogen peroxide by metal complexes of water-soluble thiacalix[4]arenetetrasulfonate on the modified anion-exchangers. AB - The catalytic activity for the decomposition of hydrogen peroxide by anion exchangers modified with metal complexes of thiacalix[4]arenetetrasulfonate (Me(n+)-TCAS[4], Me(n+)=Mn(3+), Mn(2+), Fe(3+), Co(3+), Co(2+), Cu(2+), Zn(2+) and Ni(2+)) was investigated. As a reference, calix[4]arenetetrasulfonate, calix[6]arenehexasulfonate and calix[8]areneoctasulfonate were also examined. Mn(3+)- and Fe(3+)-TCAS[4] on the modified anion-exchangers showed high catalytic activity in alkaline buffer solutions among metal complexes tested. Mn(3+)- and Fe(3+)-TCAS[4] on the modified anion-exchangers exhibited high and constant levels of catalytic activity even after having been used 5 times, and showed catalytic activity in the presence of an excess of H(2)O(2) over Mn(3+)- and Fe(3+)-TCAS[4] on the modified anion-exchangers. Only Mn(3+)-TCAS[4] on the modified anion-exchangers exhibited high catalytic activity at around a neutral pH. PMID- 14758017 TI - Simple synthesis of beta-D-glycopyranosides using beta-glycosidase from almonds. AB - Enzymatic glycosidation of twenty-one kinds of alcohols (n-hepanol, n-octanol, 2 phenylethanol, 3-phenylpropanol, 4-phenylbutanol, 5-phenylpentanol, 6 phenylhexanol, furfury alcohol, 2-pyridinemethanol, isobutanol, isopentanol, p methoxycinnamylalcohol) including secondary alcohols (isopropanol, cyclohexanol, 1-phenylethanol) and 1,omega-alkanediols (1,5-pentanediol, 1,6-hexanediol, 1,7 heptanediol, 1,8-octanediol, 1,9-nonanediol), salicyl alcohol and 4-nitrophenyl beta-D-glucopyranoside (5) using beta-glucosidase from almonds stereoselectively gave the corresponding beta-D-glucopyranosides in moderate yield. PMID- 14758018 TI - New flavonoids from Oxytropis myriophylla. AB - Eight compounds were isolated from Oxytropis myriophylla. On the basis of spectral analyses, their structures were elucidated to be (6R,9R)-roseoside (1), (6R,9S)-roseoside (2), adenosine (3), myriophylloside B (4), myriophylloside C (5), myriophylloside D (6), myriophylloside E (7), and myriophylloside F (8). Five flavonoids (4-8) were new compounds, and the three known compounds were isolated from this plant for the first time. PMID- 14758019 TI - Preparation of new nitrogen-bridged heterocycles. 55. Reinvestigation of the bromination/dehydrobromination of ethyl 3-[2-(methylthio)indolizin-3-yl]acrylate derivatives. AB - The bromination/dehydrobromination reactions of ethyl 3-[1-alkoxycarbonyl-2 (methylthio)indolizin-3-yl]acrylates were reinvestigated. Reactions of the title compounds with two equivalents of bromine, followed by heating of the resulting reaction mixture and then treatment with a base gave the unexpected dialkyl 7 bromothieno[2,3-b]indolizine-2,9-dicarboxylates, while similar reactions using benzyltrimethylammonium tribromide as a brominating agent afforded only non brominated thieno[2,3-b]indolizine derivatives, which were converted to the corresponding 7-bromo derivatives upon further treatment with bromine. PMID- 14758020 TI - Chemistry of renieramycins. Part 4.synthesis of a simple natural marine product, 6-hydroxy-7-methoxyisoquinolinemethanol. AB - 6-Hydroxy-7-methoxyisoquinolinemethanol (15) and mimosamycin (1) were recently isolated from a marine sponge, Haliclona sp. The former was prepared in ten steps from vanillin (22) in 26% overall yield using an isopropyl for phenol protection. PMID- 14758021 TI - A highly diastereoselective pinacol coupling reaction of aldehydes and ketones using low-valence niobium generated from Nb(V). AB - A simple method for the diastereoselective synthesis of racemic 1,2-diol mediated by low-valence niobium generated in situ is described. A 1,4-dioxane-toluene solvent system was found to be essential to achieve higher selectivities and to prevent other reactions of pinacols, such as deoxygenation and acetal formation. Aromatic aldehydes and ketones were converted to the corresponding pinacols with up to 97 and 85% de, respectively. PMID- 14758022 TI - A novel iridoid from Boschniakia rossica. AB - A novel iridoid, (4R)-4-hydroxymethylboschnialactone (1), has been isolated from Boschniakia rossica, together with three previously known compounds, (24S)-3beta hydroxy-24-ethylcholest-5-en-7-one (2), (24R)-3beta-hydroxy-24-ethylcholest-5-en 7-one (3) and methyl p-coumarate, using column chromatography. The structures of compounds were elucidated by spectroscopic methods. PMID- 14758023 TI - Superoxide generation by Nox1 in guinea pig gastric mucosal cells involves a component with p67(phox)-ability. AB - Nox1, a homologue of gp91(phox) subunit of the phagocyte NADPH oxidase, is responsible for spontaneous superoxide (O(2)(-)) generation in guinea pig gastric mucosal cells (GMC), but involvement of regulatory components (p67(phox), p47(phox), and Rac) which are essential in phagocytes remains unknown. Here, we aimed to figure out how Nox1 of GMC achieves an active oxidase status. GMC in primary culture show low O(2)(-) generation but acquire a 9-fold higher activity when cultured with Helicobacter pylori lipopolysaccharide (LPS), in correlation with a far increased Nox1 expression. Investigation into the O(2)(-)-generating ability of LPS-induced Nox1 in cell-free reconstitution assays showed that: 1) Nox1 is unable to generate O(2)(-) per se; 2) the combination of Nox1 with GMC cytosol is insufficient for a significant O(2)(-) generation; 3) the combination with neutrophil cytosol enables Nox1 to act like gp91(phox), i.e., to produce O(2)(-) appreciably in response to myristate stimulation; 4) Nox1 prefers NADPH to NADH under the in vitro assay with neutrophil cytosol plus myristate (K(m)=10.4 microM); 5) substitution of neutrophil cytosol by a set of recombinant cytosolic components (rp67(phox), rp47(phox), Rac2) is, however, ineffective and still requires GMC cytosol. Thus, Nox1 probably requires an additional cytosolic factor(s). In contrast, GMC cytosol enables cytochrome b(558) to generate plenty of O(2)(-), on condition that rp47(phox) is added. This result suggests that GMC cytosol contains a component with p67(phox)-ability, and also Rac, but lacks p47(phox). These data indicate that GMC Nox1 requires at least a p67(phox) counterpart and Rac to acquire NADPH oxidase activity. PMID- 14758024 TI - Preventive effect of Juzen-taiho-to on endometrial carcinogenesis in mice is based on Shimotsu-to constituent. AB - Juzen-taiho-to, a Kampo formula, originally consists of a mixture of Shimotsu-to and Shikunshi-to formulas together with two other crude ingredients. Juzen-taiho to is reported to have a preventive effect on endometrial carcinogenesis in mice. Shimotsu-to exerts an inhibitory effect on estrogen-induced expression of c-fos, interleukin (IL)-1alpha and tumor necrosis factor (TNF)-alpha in uteri of ovarectomized mice. In the present study, short- and long-term experiments were designed to determine the effects of Juzen-taiho-to and Shimotsu-to on the estrogen-related endometrial carcinogenesis in mouse uteri, associated with the expression of cyclooxygenase (COX)-1 and -2. In the short-term experiment, exposure to Juzen-taiho-to or Shimotsu-to significantly reduced estradiol-17beta (E(2))-stimulated expressions of COX-2 mRNA (p<0.05) as well as the protein. However, no effects on the expression of COX-1 were observed. Shikunshi-to did not affect COX expression. In the long-term experiment, 90 female ICR mice were given N-methyl-N-nitrosourea (MNU) into their uterine corpora. The animals were divided into four groups as follows: group 1, a diet containing 0.07% Shimotsu-to and 5 ppm E(2); group 2, a diet containing 5 ppm E(2); group 3, a diet containing 0.07% Shimotsu-to; group 4 served as a control. Exposure of Shimotsu-to reduced the incidence of MNU- and E(2)-induced endometrial adenocarcinoma and atypical hyperplasia at the termination of the experiment (30 weeks). The above findings and our previous reports suggest that Shimotsu-to is responsible for the preventive effects of Juzen-taiho-to on estrogen-related endometrial carcinogenesis in mice, through the inhibition of estrogen-related COX-2 as well as c-fos, IL-1alpha and TNF-alpha expressions. PMID- 14758025 TI - Effect of ethanol extracts of three Chinese medicinal plants with laxative properties on ion transport of the rat intestinal epithelia. AB - The effects of ethanol extracts of three Chinese medicinal plants Dahuang (Rheum palmatum L.), Badou (Croton tiglium L.), and Huomaren (Cannabis sativa L.), on ion transport of the rat intestinal epithelia were studied. Rat intestinal epithelia mounted in an Ussing chamber attached with voltage/current clamp were used for measuring changes of the short-circuit current across the epithelia. The intestinal epithelia were activated with current raised by serosal administration of forskolin 5 microM. Ethanol extracts of the three plants all augmented the current additively when each was added after forskolin. In subsequent experiments, ouabain and bumetanide were added prior to ethanol extracts of these medicinal plants to determine their effect on Na(+) and Cl(-) movement. The results suggest that ethanol extracts of the three medicinal plants may affect the Cl(-) movement more directly than Na(+) movement in the intestinal epithelial cells. The results provide evidence for the pharmacologic mechanism of the three Chinese medicinal plants on the intestinal tract. PMID- 14758026 TI - Effects of monochlorobimane on cerebral ischemia-induced damage to mitochondria. AB - A possible involvement of inhibitory effects of monochlorobimane (MCB) on the opening of mitochondrial permeability transition (MPT) pore in the cerebroprotection against the ischemic brain injury was examined. MCB (1 mM) inhibited the opening of MPT pore in vitro. Sustained cerebral ischemia was induced by injecting 900 microspheres (48 microm in diameter) into the right hemisphere of rats. At 12 to 72 h after microsphere embolism (ME), the mitochondrial activity was determined histochemically by staining cytochrome c oxidase (COX) and succinate dehydrogenase (SDH) of the brain sections. The COX and SDH stainings in the hippocampus were observed intensively in the pyramidal neurons in the CA2-3 and dentate gyrus rather than those in the CA-1 region. The staining was decreased with time after the embolism. Pretreatment with 10 microg/animal MCB 30 min prior to the embolism significantly attenuated the ME induced reduction in the staining of COX and SDH in the hippocampus, but not in the pariatal cortex. The results suggest that prevention of the opening of MPT pore by MCB may play an important role in the cerebroprotection against cerebral ischemic injury. PMID- 14758027 TI - Evaluation of the nitric oxide radical scavenging activity of manganese complexes of curcumin and its derivative. AB - Curcumin manganese complex (CpCpx) and diacetylcurcumin manganese complex (AcylCpCpx) were determined as to their effect on the nitric oxide (NO) radical scavenging in vitro method using a sodium nitroprusside generating NO system compared with their parent compound and astaxanthin, an extreme antioxidant. All compounds effectively reduced the generation of NO radicals in a dose dependent manner. They exhibited strong NO radical scavenging activity with low IC(50) values. The IC(50) values of curcumin, diacetylcurcumin, CpCpx and AcylCpCpx obtained are 20.39+/-4.10 microM, 28.76+/-1.48 microM, 9.79+/-1.50 microM and 8.09+/-0.99 microM, respectively. CpCpx and AcylCpCpx show greater NO radical scavenging than their parent compounds, curcumin and acetylcurcumin, respectively. However, the IC(50) values of curcumin and related compounds were found to be less than astaxanthin, an extreme antioxidant, with the lower IC(50) value of 3.42+/-0.50 microM. PMID- 14758028 TI - Modulation of voltage-gated Ca2+ current by 4-hydroxynonenal in dentate granule cells. AB - Although recent studies have suggested that dentate granule cells play a key role in hippocampal functions, electrophysiological properties in these cells have not been sufficiently explored. In the present study, modification of voltage-gated Ca2+ channels by 4-hydroxynonenal (4HN), a major aldehydic product of membrane lipid peroxidation, in cultured dentate granule cells was examined using the whole-cell patch clamp technique. When whole-cell voltage clamp was applied, the cells exhibited a high-voltage-activated Ca2+ current, which was totally sensitive to 30 microM Cd2+ and partially sensitive to 2 microM nifedipine. 4HN enhanced the Ca2+ current in these cells. When L-type Ca2+ channels were blocked by application of nifedipine, the enhancement was completely canceled, whereas application of omega-conotoxin-GVIA or omega-agatoxin-IVA, blockers of N- and P/Q type Ca2+ channels, respectively, had no effect. These results suggest that 4HN modulates L-type Ca2+ channels in the dentate granule cells, and thereby plays a role in the physiological and pathophysiological responses of these cells to oxidative stress. PMID- 14758029 TI - Growth-inhibitory effects of the red alga Gelidium amansii on cultured cells. AB - The objective of this study was to investigate the effects of Gelidium amansii, an edible red agar cultivated off the northeast coast of Taiwan, on the growth of two lines of cancer cells, murine hepatoma (Hepa-1) and human leukemia (HL-60) cells, as well as a normal cell line, murine embryo fibroblast cells (NIH-3T3). The potential role of G. amansii on the induction of apoptosis was also examined. The results indicated that all extracts from G. amansii, including phosphate buffered saline (PBS) and methanol extracts from dried algae as well as the dimethyl sulfoxide (DMSO) extract from freeze-dried G. amansii agar, inhibited the growth of Hepa-1 and NIH-3T3 cells, but not the growth of HL-60 cells. Annexin V-positive cells were observed in methanol and DMSO extract-treated, but not PBS extract-treated Hepa-1 and NIH-3T3 cells, suggesting that the lipid soluble extracts of G. amansii induced apoptosis. In summary, extracts of G. amansii from various preparations exhibited antiproliferative effects on Hepa-1 and NIH-3T3 cells, and apoptosis may play a role in the methanol and DMSO extract induced inhibitory effects. However, the antiproliferative effects of PBS extracts was not through apoptosis. Moreover, the growth-inhibitory effects of G. amansii were not specific to cancer cells. PMID- 14758030 TI - Interactions of phytoestrogens with estrogen receptors alpha and beta (III). Estrogenic activities of soy isoflavone aglycones and their metabolites isolated from human urine. AB - Two glucuronides (4'-O-, and 7-O-) and a glucuronyl (7-O-) sulfate (4'-O-) of genistein, two glucuronides (4'-O-, and 7-O-) and a glucuronyl (7-O-) sulfate (4' O-) of daidzein, 7-O-glucuronides of glycitein, dihydrodaidzein and O desmethylangolensin were isolated from the urine of volunteer subjects fed soy bean curds (Tofu). The estrogenic activities, i.e., i) the effect on the estrogen dependent growth of MCF-7 cells, ii) the binding ability to human estrogen receptors (hERs) alpha and beta, and iii) the effect on hER-dependent beta galactosidase induction, of these isoflavone metabolites were examined. Two synthetic isoflavone aglycones (dihydrodaidzein and O-desmethylangolensin) and four synthetic sulfates (4'-O- and 4'-, 7-di-O-) of genistein and daidzein were also studied for their estrogenic activities for the purpose of comparison. With respect to estrogenic acivity, the tested isoflavone metabolites were classified into three groups. The first group shows a very poor stimulatory effect toward the growth of MCF-7 cells, binding activity, and beta-galactosidase induction. The sulfates belong to this group. The second group shows a moderate binding activity but poor stimulation and beta-galactosidase induction. Some glucuronyl conjugates belong to this group. The last group shows a moderate stimulation and beta-galactosidase induction but poor binding activity. A mixed type of conjugates having glucuronyl and sulfony moieties belong to this group. PMID- 14758032 TI - Trypanocidal constituents in plants 4. Withanolides from the aerial parts of Physalis angulata. AB - The constituents of the aerial parts of Physalis angulata (Solanaceae) were investigated based on the plant's trypanocidal activity against epimastigotes of Trypanosoma cruzi, the etiologic agent for Chagas' disease. Four new withanolides were isolated, along with six known ones, from the active fraction. Their structures were determined by spectroscopic analysis. Trypanocidal activity against trypomastigotes, an infectious form of T. cruzi, was also estimated, as well as cytotoxic activity against human uterine carcinoma (HeLa) cells in vitro. Evaluation of trypanocidal activity using the colorimetric reagent Cell Counting Kit-8 was also examined. PMID- 14758031 TI - Antihypertensive effect of peptides from royal jelly in spontaneously hypertensive rats. AB - We have shown that Protease N treated Royal Jelly (ProRJ) and peptides from ProRJ (Ile-Tyr (IY), Val-Tyr (VY), Ile-Val-Tyr (IVY)) inhibited angiotensin I converting enzyme (ACE) activity and they have an antihypertensive effect in repeated oral administration for 28 d on spontaneously hypertensive rats (SHR). We investigated the contributive ratio of these peptides in ProRJ for antihypertensive effect in single oral administration on SHR. In single oral administration of each peptide and peptides mixture (MIX; IY, VY and IVY) at doses of 0.5, 1 and 10 mg/kg, systolic blood pressure (SBP) of SHR was reduced dose-dependently. This antihypertensive effect was held for 8 h. These results suggest that peptides contributed to the antihypertensive effect of ProRJ. And the contributive ratio of MIX in ProRJ for antihypertensive effect was computed to be about 38%. Therefore it is considered that intake of peptides, as a functional food would be beneficial for improving blood pressure in people with hypertension. PMID- 14758033 TI - Curcumin ameliorates left ventricular function in rabbits with pressure overload: inhibition of the remodeling of the left ventricular collagen network associated with suppression of myocardial tumor necrosis factor-alpha and matrix metalloproteinase-2 expression. AB - OBJECTIVE: Curcumin is a wide-spectrum cellular protector with antiinflammatory, antioxidizant, and antifibrotic effects. This study was conducted to investigate its effects on myocardial collagen remodeling in pressure overloaded rabbits. METHODS AND RESULTS: Pressure overloaded rabbits were established by partial abdominal aorta ligation. The rabbits were divided into the sham-operation group, vehicle group and curcumin group. Curcumin was administered orally at a dose of 100 mg/kg.d in 10 ml of 2.5% polyethylene glycol solution and the other 2 groups were given the same dose of polyethylene glycol solution. Compared with the vehicle group, left ventricular function in the curcumin group was significantly ameliorated, as indicated by decreased left ventricular end-diastolic pressure, left ventricle weight to body weight ratio, and the left ventricular posterior wall thickness. The collagen volume fraction in the curcumin group was also reduced. Myocardial tumor necrosis factor (TNF)-alpha and matrix metalloproteinase (MMP)-2 expression were significantly overexpressed in the vehicle group and markedly suppressed in the curcumin group at both the 4th and 8th weeks. At the end of the 8th week, the ejection fraction in the curcumin group was increased compared with that in the vehicle group. CONCLUSION: Curcumin improved left ventricular function in pressure overloaded rabbits. This might be due to inhibition of collagen remodeling associated with suppression of myocardial expression of tumor necrosis factor-alpha, and matrix metalloproteinase-2. PMID- 14758035 TI - Sweet potato acid phosphatase immobilized on glutaraldehyde-activated aminopropyl controlled-pore glass: activation, repeated use and enzyme fatigue. AB - Sweet potato acid phosphatase was covalently coupled with glutaraldehyde to aminopropyl controlled-pore glass, and used as a pre-column enzyme reactor. The immobilized enzyme reactor (IMER) was continuously operated using an automated chromatographic detection system we developed. Functional evaluation of the IMER was carried out by injecting ten samples on the same day at an injection amount of 1.25 nmol (62.5 nmol per ml) using riboflavin sodium phosphate (FMNs) as a substrate, and by prolonged use for ten months. The IMER exhibited decreased activity after repeated use for a total of 3000 samples, but about 75% of its original activity remained. The conversion rate of FMNs to riboflavin by IMER was increased from 89 to 97% by adding citrate, ethylenediaminetetraacetic acid disodium salt, etc., but especially by adding citrate. The increased conversion of FMNs to riboflavin due to the addition of citrate was probably not due to the chelation of heavy metal ions by citrate. We also investigated complex formation of acid phosphatase with the substrate FMNs using surface plasmon resonance to determine the effect of citrate on the processes of association and/or dissociation between the enzyme and substrate. Enzyme fatigue was also observed during the course of prolonged and repeated use. PMID- 14758034 TI - Evaluation of factors to decrease plasma concentration of an HIV protease inhibitor, saquinavir in ethanol-treated rats. AB - Although alcohol consumption is a factor in which the bioavailability of saquinavir (SQV) are retarded, the cause for this phenomenon remains to be uncertain. In the presence study, we examined factors to decrease plasma concentration of SQV in ethanol-treated rats. The ethanol-treated rats were prepared by making them freely access to 15% ethanol solution for 14 d (Day 14 rats). The exsorption clearance of SQV from the blood circulation to the jejunal lumen in the Day 14 rats increased by 6-fold as compared to ethanol non-treated (NT) rats. In the presence of 25 microM ketoconazole (KCZ) or 10 microM cyclosporin A (CsA) in the jejunal lumen, the plasma concentration of SQV in the portal vein increased significantly, and this effect of 10 microM CsA was superior to that of 25 microM KCZ. The biliary excretion clearance of SQV in Day 14 rats also increased by 1.8-fold as compared to that in the NT rats. The metabolic clearance rate (V(max)/K(m)) of SQV in the intestinal microsomes from the Day 14 rats increased significantly, while in the liver microsomes the V(max)/K(m) did not change. The phase II metabolism processes in the Day 14 rats based on UDP-glucuronosyltransferases and gultathion-S-tnrasferase activities were activated, however, they were not likely to be one of factors to decrease the bioavailability of oral SQV, because CYP3A activity in the liver and intestine was not activated to such an extent and SQV itself was not conjugated. These observations suggest that a main possible factor to explain the reducing effect on the SQV oral bioavailability during ethanol consumption is an enhanced efflux of SQV at the intestine and liver, where it is suggested that functional enhancement or excessive expression of P-glycoprotein is caused by ethanol consumption. PMID- 14758036 TI - Momordica charantia extracts inhibit uptake of monosaccharide and amino acid across rat everted gut sacs in-vitro. AB - The inhibitory effects of Momordica charantia extracts were studied on the uptake of glucose and tyrosine across rat everted gut sacs in vitro. The aqueous extract of the plant was found to inhibit primarily the uptake of glucose in a dose dependent manner. Uptake of tyrosine was affected at high substrate concentrations only. The extract was also found to decrease the absorptive capacity of fluid across the small intestine and sodium ions. It is hypothesized that the effects of Momordica could involve a washout of glucose from the blood stream. PMID- 14758037 TI - Expression of the soluble extracellular domain of human thrombopoietin receptor using a maltose-binding protein-affinity fusion system. AB - The thrombopoietin (TPO) receptor (Mpl) belongs to the family of ligand-dependent cytokine receptors and plays a functional role in regulating platelet production. The signaling capacity largely depends on the binding of TPO to the extracellular domains of the TPO receptor (Mpl-EC). Because the expression level of Mpl in human tissue is very low, studies on the functional and spatial characteristics of its ligand-binding sites have been limited. In the present study, we report the expression and purification of Mpl-EC as a fusion with the maltose-binding protein (MBP), designated MBP-Mpl-EC. MBP-Mpl-EC was expressed in the cytoplasm of Escherichia coli as a soluble fusion protein. Specific binding of TPO to purified MBP-Mpl-EC was demonstrated by a dot-blot assay and surface plasmon resonance. We conclude that bacterial expression of MBP-Mpl-EC yields large amounts of protein with correct folding and that it can be used for further structure and function analyses. PMID- 14758038 TI - State of chromatin sensitivity to DNase I in the rat Ig-beta/growth hormone locus determined by real-time PCR. AB - Using Ig-beta and growth hormone producing cells with liver-derived cells for controls, sensitivity of chromatin to DNase I was measured by real-time PCR at eleven targets in rat Ig-beta/growth hormone locus where four cell type-specific genes and two ubiquitously expressed genes are present in a compact 88-kb region. Chromatin situated at the promoter of actively-transcribed gene and placed at cell type-specific DNase I hypersensitive sites with enhancer activity was sensitive to DNase I. In the case of inactive gene, chromatin located in these regions was resistant to DNase I. Unexpectedly, however, chromatin placed in the transcribed intron was resistant to DNase I in two genes. DNase I sensitive chromatin was shown not to distribute locus-widely but rather to localize at the promoter and the enhancer of actively-transcribed genes in this locus. PMID- 14758039 TI - Comparative pharmacokinetic behavior of glycyrrhetic acid after oral administration of glycyrrhizic acid and Gancao-Fuzi-Tang. AB - Comparative pharmacokinetic profiles of glycyrrhetic acid (GA), glycyrrhizic acid (GL) and Gancao-Fuzi-Tang (KF) after oral administration of GL and KF were studied. Plasma samples taken from rats were acidified with acetic acid and GA was extracted with isopropanol-ethyl ether (1 : 1). Separation of GA was performed on a C(18) column with the detection wavelength set at 254 nm. The mobile phase was methanol-acetonitrile-water-acetic acid (58 : 18 : 24 : 1 v/v). The results showed that the mean residence time and area under the curve of GA in KF-administered rats were 27.6+/-1.5 h and 122.8+/-46.7 microg.h/ml respectively, which were significantly different from those in GL-administered rats (15.0+/-2.0 h and 40.9+/-9.6 microg.h/ml, respectively). The results suggest the increased effect of GA after oral administration of KF in comparison with GL. PMID- 14758040 TI - Influence of the light schedule on the toxicity of amitriptyline in chick embryos. AB - The cardiac toxicity of amitriptyline and the effect of the light schedule on it were studied in chick embryos. Fertilized eggs of White Leghorns were incubated under dark conditions and investigated, on two occasions, in the light phase and in the dark phase. Amitriptyline was injected into the air sac of fertilized eggs on the 16th day of incubation. Electrocardiograms were recorded 0 to 60 min after the injection. After the administration of amitriptyline 1 mg/egg in the light phase, the heart rate did not differ compared with that in controls. However, the heart rate was significantly decreased by the administration of amitriptyline 2.5 mg/egg and 5 mg/egg in the light phase. The heart rate was significantly decreased by the administration of amitriptyline 1 mg/egg under dark conditions. In addition, arrhythmia was produced by the administration of amitriptyline under dark conditions. These findings indicate that manipulation of the light schedule has a marked influence on the toxicity of amitriptyline in chick embryos. PMID- 14758041 TI - Oral administration of beta-cryptoxanthin induces anabolic effects on bone components in the femoral tissues of rats in vivo. AB - The effect of beta-cryptoxanthin on bone components in the femoral tissues of rats was investigated. Beta-cryptoxanthin was isolated from Satsuma mandarin (Citrus unshiu MARC.). Bone tissues were cultured for 48 h in serum-free Dulbecco's modified Eagle's medium containing either vehicle or beta cryptoxanthin (10(-7) or 10(-6) M). The presence of beta-cryptoxanthin (10(-7) or 10(-6) M) caused a significant increase in calcium content and alkaline phosphatase activity in the femoral-diaphyseal and femoral-metaphyseal tissues. These increases were completely abolished in the presence of cycloheximide (10( 6) M), an inhibitor of protein synthesis. Thus beta-cryptoxanthin had an anabolic effect on bone calcification in vitro. Moreover, beta-cryptoxanthin (10, 25, or 50 microg/100 g body weight) was orally administered once daily for 7 d to young male rats. The administration of beta-cryptoxanthin (10, 25, or 50 microg/100 g body weight) caused a significant increase in calcium content and alkaline phosphatase activity in the femoral-diaphyseal and femoral-metaphyseal tissues. Femoral-diaphyseal and femoral-metaphyseal DNA contents were significantly increased by the dose of 25 or 50 microg/100 g body weight. A significant increase in metaphyseal DNA content was also seen with the dose of 10 microg/100 g body weight of beta-cryptoxanthin. This study demonstrates that beta cryptoxanthin has an anabolic effect on bone components in rats in vitro and in vivo. PMID- 14758042 TI - Relationship between liver injury and transglutaminase activities in guinea pigs and rats. AB - We investigated the effect of transglutaminase (TGase) on in guinea pigs and rats. Serum alanine aminotransferase (ALT) level increased 1 d after CCl(4) treatment of both in guinea pigs and rats since TGaese activity was greatly higher in guinea pigs than rats. However, serum ALT level in guinea pigs was very much lower than that in rats. Liver TGase activities decreased after CCl(4) treatment in both guinea pigs and rats. However, TGase activities in the liver from guinea pigs were higher than that from rats. Decreased TGase activities by CCl(4) in the liver from guinea pigs and rats were significantly recovered by retinoic acid treatment that was reported to increase TGase. Degree of recovery of serum ALT level by retinoic acid in rats was larger than in guinea pigs. These results suggested that the distinction of the effect of retinoic acid on serum ALT level in CCl(4)-treated animals was due to the different TGase activity that increased membrane stability. PMID- 14758043 TI - Influence of receptor number on the cAMP response to forskolin in Chinese hamster ovary cells transfected with human beta2-adrenoceptor. AB - We examined the basal adenosine 3',5'-cyclic monophosphate (cAMP) levels and forskolin-induced cAMP accumulation in cultured Chinese hamster ovary cells (CHO) expressing different levels of human beta(2)-adrenoceptors. Both the basal and forskolin-induced cAMP accumulation in the cells that express higher density of beta(2)-adrenoceptors (CHO-beta(2)/H; 560 fmol/mg protein) were larger than those in the cells that express lower density of beta(2)-adrenoceptors (CHO-beta(2)/L; 270 fmol/mg protein). In addition, isoproterenol-induced cAMP accumulation was also augmented as the number of beta(2)-adrenoceptors was increased. ICI 118,551, a selective beta(2)-adrenoceptor antagonist with inverse agonist properties, decreased all the levels of cAMP observed in both cell lines. These results suggest that the agonist-independent (constitutive) activity of beta(2) adrenoceptors plays a key role in the control of forskolin-induced cAMP accumulation. PMID- 14758044 TI - No effects of chlorophyllin on IQ (2-amino-3-methylimidazo[4,5-f]-quinoline) genotoxicity and -DNA adduct formation in Drosophila. AB - Previously we demonstrated that chlorophyllin suppressed the genotoxicities of many carcinogens. However, the genotoxicity of IQ (2-amino-3-methylimidazo[4,5 f]quinoline), a carcinogenic heterocyclic amine, was not suppressed in Drosophila. On the contrary, it has been reported that chrolophyllin suppressed the genotoxicity of IQ in rodents, rainbow trout and Salmonella. We demonstrated that the chlorophyllin-induced suppression of MeIQx (2-amino-3,8 dimethylimidazo[4,5-f]quinoxaline)-genotoxicity was associated with a decrease in MeIQx-DNA adduct formation in Drosophila larval DNA. MeIQx represents another type of heterocyclic amine and is similar to IQ in structure. In this study we utilized (32)P-postlabeling to examine whether chlorophyllin reduced IQ-DNA adduct formation in Drosophila DNA in the same way as MeIQx. The results revealed that the formation of IQ-DNA adducts was unaffected by treatment with chlorophyllin. This was consistent with the absence of any inhibitory effect on genotoxicity as observed in the Drosophila repair test. These results suggest that IQ-behavior in Drosophila is not affected by chlorophyllin, indicating that the process of IQ-DNA adduct formation followed by expression of genotoxicity in Drosophila may be different from that in other organisms. PMID- 14758045 TI - 2',6'-dimethylphenylalanine (Dmp) can mimic the N-terminal Tyr in opioid peptides. AB - Substitution of 2',6'-dimethyltyrosine (Dmt) for the N-terminal Tyr in opioid peptides has recently been shown to be a promising tool for improving opioid receptor affinity and biological activity. We have also demonstrated that another unnatural amino acid, 2',6'-dimethylphenylalanine (Dmp), is not only an excellent substitute for Phe at position 3 but also can mimic the aromatic N-terminal Tyr residue in a micro opioid receptor-selective dermorphin analogue (YRFB: Tyr-D-Arg Phe-betaAla-NH(2)). To further evaluate the value of Dmp in opioid peptides, we investigated Dmp(1)-substituted analogues of the delta receptor ligands, deltorphin II (DLT: Tyr-D-Ala-Phe-Glu-Val-Val-Gly-NH(2)) and enkephalin (ENK: Tyr Gly-Gly-Phe-Leu). In the receptor binding assay, both [Dmp(1)]DLT and [Dmp(1)]ENK bound to the delta-receptor with high affinity and selectivity, and were nearly as effective as the parent peptides. The potency of the Dmp(1)-peptides on the MVD and GPI assays correlated well with the receptor binding affinity data. These results are in contrast to the tendency of corresponding Dmt(1)-analogues to have poor receptor selectivity. Taken together with the results with YRFB, we conclude that the Dmp(1)-peptide is superior to the corresponding Dmt(1)-peptide in its receptor selectivity. [Dmp(1)]DLT and [Dmp(1)]YRFB may serve as pharmacological tools for the studies of ligand recognition and opioid receptor signal transduction. PMID- 14758046 TI - Suppressive activity of the fruit of Momordica charantia with exercise on blood glucose in type 2 diabetic mice. AB - The antidiabetic activity of Momordica charantia L. (Cucurbitaceae) with exercise was investigated in KK-Ay mice, an animal model with type 2 diabetes with hyperinsulinemia. The water extract of the fruit of Momordica charantia L. (MC) with exercise reduced the blood glucose of KK-Ay mice 5 weeks after oral administration (p<0.001), and also significantly lowered the plasma insulin of KK Ay mice under similar conditions (p<0.01). The blood glucose of MC with exercise is lower than that of MC only or exercise only 5 weeks after the administration. MC with exercise decreased blood glucose in a glucose tolerance test. These results suggest that MC with exercise is useful for type 2 diabetic cure. PMID- 14758047 TI - Antiproliferative activity of the main constituents from Phyllanthus emblica. AB - Eighteen main compounds, including four norsesquiterpenoids (1-4) and 14 phenolic compounds (5-18) isolated previously from Phyllanthus emblica, together with a main constituent, proanthocyanidin polymers (19) identified at this time from the roots, were estimated for their antiproliferative activities against MK-1 (human gastric adenocarcinoma), HeLa (human uterine carcinoma), and B16F10 (murine melanoma) cells using an MTT method. All of the phenolic compounds including the major components 5-8 from the fruit juice, 8, 9, and 12 from the branches and leaves, and 19 from the roots showed stronger inhibition against B16F10 cell growth than against HeLa and MK-1 cell growth. Norsesquiterpenoid glycosides 3 and 4 from the roots exhibited significant antiproliferative activities, although their aglycon 1 and monoglucoside 2 showed no inhibitory activity against these tumor cells. PMID- 14758048 TI - Effect of the water-soluble and non-dialyzable fraction isolated from Senso (Chan Su) on lymphocyte proliferation and natural killer activity in C3H mice. AB - We found lymphocyte proliferating substances in the water-soluble and non dialyzable fraction prepared from the crude drug Senso (Chan Su). The effect of this fraction was increased by affinity chromatography using the concanavalin A agarose. By analyzing the fraction using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and lectin blotting, we estimated that one of the active substances of this fraction is a glycoprotein that has about 13 kDa of molecular weight and D-mannose within the molecule. The purified fraction increased the IL-2 and the IL-12 level in the supernatant of spleen cell culture, and increased the natural killer activity of spleen lymphocyte in C3H/HeN mice. These results show that Senso contains immunopotentiating substances that may serve as an immunomodulator in an organism. PMID- 14758049 TI - Pharmacokinetics of lopinavir after administration of Kaletra in healthy Japanese volunteers. AB - The pharmacokinetic parameters of lopinavir (LPV) were examined by administering Kaletra (LPV+ritonavir) to 8 healthy Japanese volunteers both in the fasting and postprandial conditions. LPV showed a biphasic decline, which was slower in the initial phase and became more rapid in the later phase. The behavior of LPV in the initial phase could be modeled using a one-compartment model with first-order absorption. In the fasting study, calculations based on the pharmacokinetic model revealed that the time to reach the maximum concentration (T(max)), maximum concentration (C(max)), half-life (T(1/2)), lag time, apparent volume of distribution (Vd/F) and oral clearance (Cl/F) were 3.2+/-1.0 h, 6.9+/-1.9 microg/ml, 10.0+/-3.7 h, 0.71+/-0.32 h, 51.0+/-12.4 l and 4.2+/-2.6 l/h, respectively. On the other hand, in the postprandial study, the calculated T(max), C(max), T(1/2), lag time, Vd/F and Cl/F were 5.6+/-2.0 h, 7.6+/-1.8 microg/ml, 16.7+/-7.0 h, 2.35+/-0.78 h, 48.0+/-15.9 l and 2.1+/-0.6 l/h, respectively. The values for the area under the curve for data collected over a 24-h period (AUC(24 h)) in the fasting and postprandial studies were 86.0+/-27.7 and 102.1+/-31.0 microg.h/ml, respectively. The T(1/2) had a tendency to be prolonged after food intake, but there were 2 cases with shortened T(1/2). Food intake prolonged the lag time 3-fold and as a result, the postprandial T(max) was 2 times longer. PMID- 14758050 TI - Fucoidan modulates the effect of transforming growth factor (TGF)-beta1 on fibroblast proliferation and wound repopulation in in vitro models of dermal wound repair. AB - Aberrant wound healing, either causing scarring or chronic wounds, is a significant cause of morbidity. There is therefore, considerable interest in agents which can modulate certain aspects of the wound healing process. Fucoidans, sulphated polyfucose polysaccharides which may be extracted from Fucus spp., have been shown to modulate the effects of a variety of growth factors through mechanisms thought to be similar to the action of heparin. We investigated the interaction between two commercial preparations of fucoidan and transforming growth factor (TGF)-beta(1). These preparations of fucoidan, as well as heparin, inhibited fibroblast proliferation at concentrations from 0.01 to 100 mg/ml. The anti-proliferative effects of 1 ng/ml TGF-beta(1) on dermal fibroblasts were abrogated by fucoidan preparation F7 when used at concentrations over 1 mg/ml. In a three dimensional in vitro model of wound repair, the fibroblast populated collagen lattice or "dermal equivalent", TGF-beta(1) reduced the rate of fibroblast repopulation of a wound defect created by punch biopsy. Addition of fucoidan to the model in the presence of TGF-beta(1) increased the rate of fibroblast repopulation of the wound and at 10 mg/ml of fucoidan the number of cells which had migrated into the wounded defect was similar to that of control cultures. These data suggest that fucoidan has properties which may be beneficial in the treatment of wound healing. PMID- 14758051 TI - Effect of carboxylation of N-terminal phenylalanine of (111)in-DTPA (diethylenetriaminepentaacetic acid)-octreotide on accumulation of radioactivity in kidney. AB - For purpose of reducing renal accumulation of radioactivity of a known radiopharmaceutical agent, i.e., (111)In-DTPA (diethylenetriaminepentaacetic acid)-D-Phe(1)-octreotide, a derivative in which p-carboxy-L-phenylalanine is substituted for D-Phe(1) was synthesized. Biodistribution study of the resultant compound having carboxy-substituted L-Phe(1) revealed that the renal accumulation was significantly lower than that of control compound having unsubstituted L Phe(1), demonstrating that the presence of negative charge on the N-terminal amino acid of octreotide is effective to reduce the renal accumulation. This effect can be attributed to the reduction of lipophilicity and also the repulsive force arisen from the negative charge of renal brush border membrane. PMID- 14758052 TI - Neuroendocrine aspects of chronic fatigue syndrome. AB - Chronic fatigue syndrome (CFS) is a serious health concern affecting over 800000 Americans of all ages, races, socioeconomic groups and genders. The etiology and pathophysiology of CFS are unknown, yet studies have suggested an involvement of the neuroendocrine system. A symposium was organized in March 2001 to explore the possibility of an association between neuroendocrine dysfunction and CFS, with special emphasis on the interactions between neuroendocrine dysfunction and other abnormalities noted in the immune and autonomic nervous systems of individuals with CFS. This paper represents the consensus of the panel of experts who participated in this meeting. PMID- 14758053 TI - Expression of tyrosine hydroxylase in lymphocytes and effect of endogenous catecholamines on lymphocyte function. AB - OBJECTIVES: To comprehend the changes and significance of the endogenous catecholamines in the immune system, we explored the synthesis of catecholamines by lymphocytes in various lymphoid organs and in different activated states, and the effect of the endogenous catecholamines synthesized by lymphocytes on the function of the lymphocytes themselves. METHODS: Immunohistochemistry for lymphoid organs (mesenteric lymph nodes, spleen and thymus) and lymphocytes was used to observe their expression of tyrosine hydroxylase (TH), an initial rate limiting enzyme of the catecholamine synthesis. The contents of catecholamines, including norepinephrine (NE), dopamine (DA) and epinephrine (E), in lymphocytes were tested by means of high-performance liquid chromatography with electrochemical detection. Western blot was used to examine the character and relative quantity of TH-stained protein in lymphocytes, lymph nodes and adrenal medullary tissue. The effect of alpha-methyl-P-tyrosine (alpha-MT), an inhibitor of TH activity, on concanavalin A (Con A)-induced interleukin-2 (IL-2) production was determined by MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] assay. RESULTS: TH-positive cells were found in the three examined lymphoid organs, but the lymph nodes had the highest and the thymus had the lowest density. Both TH expression and the contents of NE, DA and E in the Con A activated lymphocytes were markedly increased in comparison with those in the nonactivated lymphocytes. A band with TH immunoreactivity was seen in the extracts from either Con A-activated lymphocytes or nonactivated cells and the molecular weight of the protein was 59.4 +/- 0.3 kD. However, the relative quantity of the protein was notably higher in the activated lymphocytes than in the nonactivated cells. As a positive control, a similar band of TH immunoreactivity in the adrenal medullary tissue was also obtained. Alpha-MT at the doses of 10(-11), 10(-10) and 10(-9) M was found to significantly facilitate the Con A-induced IL-2 production. CONCLUSIONS: These results suggest that lymphocytes can synthesize catecholamines and their synthesis levels may increase in the activated state, and that endogenous catecholamines synthesized by the lymphocytes can regulate the function of the lymphocytes themselves. PMID- 14758054 TI - Regulation of leukocyte function-associated antigen 1-mediated adhesion by somatostatin and substance P in mouse spleen cells. AB - BACKGROUND: Interaction of the integrin leukocyte function-associated antigen (LFA)-1 (CD11a/CD18) with its ligands, the intercellular adhesion molecules (ICAM)-1, -2, and -3 (CD54, CD102, and CD50), is pivotal to many leukocyte adhesion events. METHOD: To define the mechanism of the movement of leukocytes to the inflammatory site by somatostatin (SOM) and substance P (SP), we examined the expression of the adhesion molecule LFA-1 and inside-out signals for integrins, protein kinase C (PKC), Ras, Rap1, and phosphoinositide (PI) 3-kinase, in anti CD3-, anti-CD3+SOM-, anti-CD3+SP-stimulated or unstimulated spleen cells. RESULTS: SOM caused down-regulation of LFA-1 mRNA translation as well as of adhesion-stimulating molecules such as Rap1, Ras, and PI 3-kinase. On the other hand, SP slightly induced LFA-1 mRNA translation and activation signals for integrins. The early-phase alteration of LFA-1 mRNA translation after 3 h of culture may be due to the changes of CD8+ T cells rather than changes of CD4+ cells. In adhesion assays, SOM significantly decreased cell adhesion (p < 0.05). CONCLUSION: These data suggest that SOM treatment of spleen cells, especially in CD8+ T cells, leads to downregulation of LFA-1 mRNA translation, inside-out signaling molecules for integrins (Ras, Rap1 and PI 3-kinase, but not PKC), and consequently to a decrease in the LFA-1-mediated adhesion to ICAM-1. PMID- 14758055 TI - Effects of cold stress on spleen cell proliferation and cytokine production during chronic Toxoplasma gondii infection. AB - BACKGROUND: Cell-mediated immunity is critical for controlling infection and preventing reactivation during the chronic phase of Toxoplasma gondii infection. In people suffering from AIDS, T. gondii is one of the major opportunistic infectious agents. Mechanisms regulating rapid development of clinical signs in previously asymptomatic patients remain unclear; however, cofactors such as stress are suspected to play a role in the susceptibility to opportunistic infections. OBJECTIVE: This study examined the role of cold stress (CS) in splenocyte function during chronic T. gondii infection. METHODS: Control mice and mice previously infected orally with T. gondii were subjected to CS during the chronic phase (CSchr), i.e. 90 days after infection, and in vitro cell proliferation and cytokine production were measured before (day 0) and 1, 15 and 25 days after CSchr. Splenocyte proliferation and cytokine production were measured after in vitro stimulation with concanavalin A (Con-A), anti-CD3 antibody (A-CD3) and Toxoplasma lysate antigen. RESULTS: CSchr enhanced splenocyte proliferation in cells stimulated with Con-A and A-CD3, but it suppressed proliferation in cells stimulated with T. gondii antigens. Increased levels of interferon (IFN)-gamma were detected independent of the type of stimulation after CSchr and remained high throughout the experiment. CS had similar results in noninfected animals. CONCLUSION: Although an overall increase in splenocyte function occurred after nonspecific stimulation, CS suppressed primed spleen cells from responding to T. gondii antigens which could lead to reactivation of latent infection. The increase in IFN-gamma after CSchr could be a result of spleen cells being primed by released parasites by this stressor. IFN gamma is critical in the control of parasite reactivation. PMID- 14758056 TI - The effects of a switch-off response accompanied by hypothalamically induced restlessness on immunoendocrinological changes in cats. AB - Electrical stimulation of the anterior hypothalamus in cats elicits a behavior called restlessness. When a switch is available for the cats to shut off the electrical stimulation, the cats learn to turn off the stimulation (switch-off response; SOR). In this study, we examined the relationship between the SOR and immunoendocrinological alterations. First of all, an escapable stimulation, in which cats could turn off the stimulation, was applied (escapable condition; EC). One month later, inescapable stimulation was delivered under the same conditions except for the fact that the cats could not turn off the stimulation (inescapable condition; IC). A behavioral analysis revealed that unstable patterns of behavior and a reduction in motor activity were observed in IC compared with those in EC. Furthermore, no significant changes in peripheral leukocytes were observed, while plasma epinephrine and cortisol transiently increased after the series of stimulations, but immediately decreased after the end of the stimulation in EC. On the other hand, there was a greater and prolonged increase in the number of peripheral granulocytes and the plasma levels of epinephrine and cortisol from 1 to 2 h after the stimulation until the end of the experiment in IC. Regarding the number of peripheral lymphocytes, CD4+ or CD8+ lymphocytes and the CD4+ to CD8+ ratio, no significant differences were found between EC and IC. These results suggest that the inability to escape from the aversive stimulation caused a decrease in movement and a prolonged alteration of the immune and endocrine systems, as is often observed in learned helplessness. PMID- 14758057 TI - Splenic denervation suppresses mRNA gene expression and protein production of IL 1beta and IL-6 by peritoneal macrophages in both Trypanosoma brucei brucei infected and non-infected rats. AB - OBJECTIVE: To test the hypothesis that the nervous system participates in modulating the immune response during experimental African trypanosomiasis caused by Trypanosoma brucei brucei. METHODS AND RESULTS: Using in situ hybridization and immunochemistry, we studied the effects of splenic sympathectomy on mRNA gene expression and protein production of IL-1beta and IL-6 in splenic and peritoneal macrophages (PMPhi) of Sprague-Dawley rats infected with T. brucei brucei and non infected rats. The enhancements of mRNA gene expression and production of IL 1beta and IL-6 by peritoneal macrophages were significantly suppressed by the splenic sympathectomy in both infected and non-infected rats. CONCLUSIONS: Our data indicate a probably stimulatory role of the sympathetic nervous system during the host immune response in both normal and T. brucei brucei-infected rats. PMID- 14758058 TI - Differential effects of light/dark recombinant human prolactin administration on the submaxillary lymph nodes and spleen activity of adult male mice. AB - OBJECTIVES: Day/night variations in cellularity, percentage of CD4+, CD8+ and double-positive (CD4+-CD8+) lymphocytes, lipopolysaccharide (LPS)- and concanavalin A (Con A)-induced lymphocyte proliferation and natural killer (NK) activity, and the effect of timed administration of recombinant human prolactin (h-PRL) on the above-mentioned parameters were investigated in the submaxillary lymph nodes and spleen of adult male mice. RESULTS: In controls, the percentage of CD4+, double-positive lymphocytes, LPS- or Con A-induced blastogenic proliferation and NK activity in the spleen differ during the dark phase as compared to the light phase. When administered during the dark period, h-PRL induced immunosuppresion in the percentage of CD4+, double-positive (CD4+-CD8+) lymphocytes. Con A- and LPS-induced lymphocyte proliferation and NK activity as compared to untreated controls. When h-PRL was administered during the light period, the cellularity increased, and h-PRL was immunosuppressive in Con A- and LPS-induced lymphcoyte proliferation and NK activity as compared to controls. Moreover, in control submaxillary lymph nodes the cellularity, percentage of CD8+, double-positive lymphocytes, blastogenic proliferation in the presence of Con A and LPS and NK activity differ when comparing the dark with the light phase. When administered during the dark period h-PRL induced immunosuppression in the percentage of double-positive (CD4+-CD8+) lymphocytes, Con A- and LPS induced lymphocyte proliferation as compared to controls. When h-PRL is administered during the light period, no effects were observed. CONCLUSIONS: These results indicate the existence of differential day/night variations in the cellular immune response depending upon the lymphoid organ considered. Because of the administration of h-PRL a differential modulation of this circadian variation was also observed. PMID- 14758059 TI - IL-10 inhibits nitric oxide synthesis in murine uterus. AB - OBJECTIVES: Recent reports point to a role for the nitric oxide/nitric oxide synthase (NO/NOS) system in implantation. It has been suggested that inducible NOS expressed at peri-implantation would lead to enhanced NO production, which could promote the attachment of the blastocyst. Short-term administration of NO donors during the pre-implantation period reduced the pregnancy rate in a dose dependent manner. Thus, it is thought that optimal levels of NO are critical for embryo implantation, so regulation of NOS must be crucial. Taking this into consideration, interleukin-10 (IL-10), synthesized and secreted by the embryo, could be modulating NOS during implantation. In this study we have investigated the in vitro effect of IL-10 on NOS in the uterus. METHODS: To determine the effect of IL-10, slices of uterus from estrogenized mice were pre-incubated for 60 min with different concentrations of IL-10 and NOS activity was measured. RESULTS: IL-10 (50 and 100 ng/ml in vitro) diminished NOS activity. The in vivo administration of lipopolysaccharide (LPS; 8 mg/kg) significantly increased the conversion of arginine into citrulline. This effect was abolished after 60 min of preincubation with IL-10 (100 ng/ml). The stimulatory effect of LPS and estrogen on NOS activity is exerted on the Ca-independent isoform and IL-10 in vitro abolished this increase. We observed that the uterus of pregnant mice on day 5 of gestation synthesized NO. This production was significantly inhibited by preincubation with IL-10 (100 ng/ml). CONCLUSIONS: This report demonstrates that IL-10 is capable of inhibiting NO synthesis in estrogenized, LPS-treated and pregnant rat uterus. PMID- 14758060 TI - State-dependent effects of Na channel noise on neuronal burst generation. AB - We explore the effects of stochastic sodium (Na) channel activation on the variability and dynamics of spiking and bursting in a model neuron. The complete model segregates Hodgin-Huxley-type currents into two compartments, and undergoes applied current-dependent bifurcations between regimes of periodic bursting, chaotic bursting, and tonic spiking. Noise is added to simulate variable, finite sizes of the population of Na channels in the fast spiking compartment. During tonic firing, Na channel noise causes variability in interspike intervals (ISIs). The variance, as well as the sensitivity to noise, depend on the model's biophysical complexity. They are smallest in an isolated spiking compartment; increase significantly upon coupling to a passive compartment; and increase again when the second compartment also includes slow-acting currents. In this full model, sufficient noise can convert tonic firing into bursting. During bursting, the actions of Na channel noise are state-dependent. The higher the noise level, the greater the jitter in spike timing within bursts. The noise makes the burst durations of periodic regimes variable, while decreasing burst length duration and variance in a chaotic regime. Na channel noise blurs the sharp transitions of spike time and burst length seen at the bifurcations of the noise-free model. Close to such a bifurcation, the burst behaviors of previously periodic and chaotic regimes become essentially indistinguishable. We discuss biophysical mechanisms, dynamical interpretations and physiological implications. We suggest that noise associated with finite populations of Na channels could evoke very different effects on the intrinsic variability of spiking and bursting discharges, depending on a biological neuron's complexity and applied current dependent state. We find that simulated channel noise in the model neuron qualitatively replicates the observed variability in burst length and interburst interval in an isolated biological bursting neuron. PMID- 14758061 TI - Effects of the activity of the internal globus pallidus-pedunculopontine loop on the transmission of the subthalamic nucleus-external globus pallidus-pacemaker oscillatory activities to the cortex. AB - Resting tremor is the most specific sign for idiopathic Parkinson' disease. It has been proposed that parkinsonian tremor results from the activity of the central oscillators. One of the hypotheses, which have been proposed about the possible principles underlying such central oscillations, is the subthalamic nucleus (STN)-external globus pallidus (GPe)-pacemaker hypothesis. Activity from the central oscillator is proposed to be transmitted via trans-cortical pathways to the periphery. A computational model of the basal ganglia (BG) is proposed for simulating the effects of the internal globus pallidus (GPi)-pedunculopontine (PPN) loop activity on the transmission of the STN-GPe-pacemaker oscillatory activities to the cortex, based on known anatomy and physiology of the BG. According to the result of the simulation, the GPi-PPN loop activity can suppress the transmission of the STN-GPe-pacemaker oscillatory activities to the cortex. This suppressive effect is controlled by various factors such as the strength of the synaptic connection from the PPN to the GPi, the strength of the synaptic connection from the GPi to the PPN, the spontaneous tonic activities of the GPi and PPN, the direct excitatory projections from the STN to the PPN, the frequency of the STN oscillatory burst activity, the duration of the STN burst, and the maximum T-type calcium channel conductance in the type-I PPN neurons. PMID- 14758062 TI - Optimal stimulus coding by neural populations using rate codes. AB - We create a framework based on Fisher information for determining the most effective population coding scheme for representing a continuous-valued stimulus attribute over its entire range. Using this scheme, we derive optimal single- and multi-neuron rate codes for homogeneous populations using several statistical models frequently used to describe neural data. We show that each neuron's discharge rate should increase quadratically with the stimulus and that statistically independent neural outputs provides optimal coding. Only cooperative populations can achieve this condition in an informationally effective way. PMID- 14758063 TI - Optimization of input patterns and neuronal properties to evoke motor neuron synchronization. AB - The study used a computational approach to identify combinations of synaptic input timing and strength superimposed on a variety of active dendritic conductances that could evoke similar levels of motor unit synchronization in model motor neurons. Two motor neurons with low recruitment thresholds but different passive properties were modeled using GENESIS software. The timing and strength of synaptic inputs and the density of dendritic ion channels were optimized with a genetic algorithm to produce a set of target discharge times. The target times were taken from experimental recordings made in a human subject and had the synchronization characteristics that are commonly observed in hand muscles. The main finding was that the two parameters with the highest association to output synchrony were the ratio of inward-to-outward ionic conductances (r = 0.344; P = 0.003) and the degree of correlation in inhibitory inputs (r = 0.306; P = 0.009). Variation in the amount of correlation in the excitatory input was not positively correlated with variation in output synchrony. Further, the variability in discharge rate of the model neurons was positively correlated with the density of N -type calcium channels in the dendritic compartments (r = 0.727; P < 0.001 and r = 0.533; P < 0.001 for the two cells). This result suggests that the experimentally observed correlation between discharge variability and synchronization is caused by an increase in fast inward ionic conductances in the dendrites. Given the moderate level of correlation between output synchrony and each of the model parameters, especially at moderate levels of synchrony (E < 0.09 and CIS < 1.0), the results suggest caution in ascribing mechanisms to observations of motor unit synchronization. PMID- 14758064 TI - An analytical model for the "large, fluctuating synaptic conductance state" typical of neocortical neurons in vivo. AB - A model of in vivo-like neocortical activity is studied analytically in relation to experimental data and other models in order to understand the essential mechanisms underlying such activity. The model consists of a network of sparsely connected excitatory and inhibitory integrate-and-fire (IF) neurons with conductance-based synapses. It is shown that the model produces values for five quantities characterizing in vivo activity that are in agreement with both experimental ranges and a computer-simulated Hodgkin-Huxley model adapted from the literature (Destexhe et al. (2001) Neurosci. 107(1): 13-24). The analytical model builds on a study by Brunel (2000) (J. Comput. Neurosci. 8: 183-208), which used IF neurons with current-based synapses, and therefore does not account for the full range of experimental data. The present results suggest that the essential mechanism required to explain a range of data on in vivo neocortical activity is the conductance-based synapse and that the particular model of spike initiation used is not crucial. Thus the IF model with conductance-based synapses may provide a basis for the analytical study of the "large, fluctuating synaptic conductance state" typical of neocortical neurons in vivo. PMID- 14758065 TI - A dynamical model of fast cortical reorganization. AB - In this work we study the connection between some dynamic effects at the synaptic level and fast reorganization of cortical sensory maps. By using a biologically plausible computational model of the primary somatosensory system we obtained simulation results that can be used to relate the dynamics of the interactions of excitatory and inhibitory neurons to the process of somatotopic map reorganization immediately after peripheral lesion. The model consists of three regions integrated into a single structure: tactile receptors representing the glabrous surface of the hand, ventral posterior lateral nucleus of the thalamus and area 3b of the primary somatosensory cortex, reproducing the main aspects of the connectivity of these regions. By applying informational measures to the simulation results of the dynamic behavior of AMPA, NMDA and GABA synaptic conductances we draw some conjectures about how the several neuronal synaptic elements are related to the initial stage of the digit-induced reorganization of the hand map in the somatosensory cortex. PMID- 14758066 TI - Introduction to galectins. AB - Good evidence suggest roles of galectins in cancer, immunity and inflammation, and development, but a unifying picture of their biological function is lacking. Instead galectins appear to have a particularly diverse, bewildering but intriguing array of activities both inside and outside cells--"clear truths and mysteries are inextricably twined". Fortunately this has not discouraged but rather enthused a large number of good galectin researchers, some of which have contributed to this special issue of Glycoconjugate Journal to provide a personal, critical status of the field. Here we will give a brief introduction to the galectins as a protein family with some comments on nomenclature. PMID- 14758067 TI - Galectin genes: regulation of expression. AB - In this review we have summarized the more recent studies on the expression of mammalian galectins. One interesting observation that can be made is that in most of microarrays and/or differential display analysis performed in recent years one or more galectins have been picked up. From a critical evaluation of the pertinent studies the main conclusion that can be drawn is that, although it is not yet clear whether the 14 galectins identified so far have functions in common, a striking common feature of all galectins is the strong modulation of their expression during development, differentiation stages and under different physiological or pathological conditions. This suggests that the expression of different galectins is finely tuned and possibly coordinated. In spite of these observations it is rather unexpected that very few studies have been performed on the molecular mechanisms governing the activity of galectin genes. PMID- 14758068 TI - The speciation of conger eel galectins by rapid adaptive evolution. AB - Many cases of accelerated evolution driven by positive Darwinian selection are identified in the genes of venomous and reproductive proteins. This evolutional phenomenon might have important consequences in their gene-products' functions, such as multiple specific toxins for quick immobilization of the prey and the establishment of barriers to fertilization that might lead to speciation, and in the molecular evolution of novel genes. Recently, we analyzed the molecular evolution of two galectins isolated from the skin mucus of conger eel (Conger myriaster), named congerins I and II, by cDNA cloning and X-ray structural analysis, and we found that they have evolved in the rapid adaptive manner to emergence of a new structure including strand-swapping and a unique new ligand binding site. In this review article we summarize and discuss the molecular evolution, especially the rapid adaptive evolution, and the structure-function relationships of conger eel galectins. PMID- 14758069 TI - Thermodynamic binding studies of galectin-1, -3 and -7. AB - The carbohydrate binding specificities of the galectin family of animal lectins has been the source of intense recent investigations. Isothermal titration microcalorimetry (ITC) provides direct determination of the thermodynamics of binding of carbohydrates to lectins, and has provided important insights into the fine carbohydrate binding specificities of a wide number of plant and animal lectins. Recent ITC studies have been performed with galectin-1, galectin-3 and galectin-7 and their interactions with sialylated and non-sialylated carbohydrates. The results show important differences in the specificities of these three galectins toward poly-N-acetyllactosamine epitopes found on the surface of cells. PMID- 14758071 TI - Galectin-1 plays essential roles in adult mammalian nervous tissues. Roles of oxidized galectin-1. AB - Previous data have suggested that galectin-1 is expressed widely in nervous tissues at embryonic stages but becomes restricted mainly to peripheral nervous tissues with maturation. Though the expression is intense in adult mammalian peripheral neurons, there had been no report about functions of galectin-1 there. Recently we discovered a factor that enhanced peripheral axonal regeneration. The factor was identified as oxidized galectin-1 with three intramolecular disulfide bonds and showed no lectin activity. Oxidized recombinant human galectin-1 (rhGAL 1/Ox) showed the same nerve growth promoting activity at very low concentrations (pg/ml). rhGAL-1/Ox at similarly low concentration was also effective in in vivo experiments of axonal regeneration. Moreover, the application of functional anti rhGAL-1 antibody strongly inhibited the axonal regeneration in vivo as well as in vitro. Since galectin-1 is expressed in the regenerating sciatic nerves as well as in both sensory neurons and motor neurons, these results suggest that galectin 1 is secreted into the extracellular space to be oxidized, and then, in its oxidized form, to regulate initial repair after axotomy. The administration of oxidized galectin-1 effectively promoted functional recovery after sciatic nerve injury in vivo. Oxidized galectin-1, hence, appears to play an important role in promoting axonal regeneration, working as a kind of cytokine, not as a lectin. Recent reports indicated additional roles of cytosolic galectin-1 in neural diseases, such as ALS. Furthermore galectin-1 has been proved to be a downstream target of DeltaFosB. In hippocampus of rat brain, expression of DeltaFosB is induced immediately after ischemia-reperfusion, suggesting that galectin-1 may also play important roles in central nervous system after injury. PMID- 14758070 TI - Galectin-1: a bifunctional regulator of cellular proliferation. AB - Galectin-1 has demonstrated a diverse range of activities in relation to cell survival and proliferation. In different circumstances, it acts as a mitogen, as an inhibitor of cell proliferation, and as a promoter of cellular apoptosis. Many of these activities, particularly the mitogenic and apoptotic responses, follow from the interaction of galectin-1 with cell-surface beta-galactoside ligands, but there is increasing evidence for protein-protein interactions involving galectin-1, and for a beta-galactoside-independent cytostatic mechanism. The bifunctional nature of galectin-1, in conjunction with other experimental variables, makes it difficult to assess the overall outcomes and significance of the growth-regulatory actions in many previous investigations. There is thus a need for well-defined experimental cross-correlation of observations, for which specific loss-of-function galectin-1 mutants will be invaluable. Unsurprisingly, in view of this background, the interpretation of the actions of galectin-1 in developmental situations, both normal and neoplastic, is often very complex. PMID- 14758072 TI - Galectin 9 is the sugar-regulated urate transporter/channel UAT. AB - UAT, also designated galectin 9, is a multifunctional protein that can function as a urate channel/transporter, a regulator of thymocyte-epithelial cell interactions, a tumor antigen, an eosinophil chemotactic factor, and a mediator of apoptosis. We review the evidence that UAT is a transmembrane protein that transports urate, describe our molecular model for this protein, and discuss the evidence from epitope tag and lipid bilayer studies that support this model of the transporter. The properties of recombinant UAT are compared with those of urate transport into membrane vesicles derived from proximal tubule cells in rat kidney cortex. In addition, we review channel functions predicted by our molecular model that resulted in the novel finding that the urate channel activity is regulated by sugars and adenosine. Finally, the presence and possible functions of at least 4 isoforms of UAT and a closely related gene hUAT2 are discussed. PMID- 14758073 TI - Understanding the biochemical activities of galectin-1 and galectin-3 in the nucleus. AB - Nuclear extracts (NE), capable of carrying out splicing of pre-mRNA, contain galectin-1 and galectin-3. NE depleted of galectins-1 and -3 concomitantly lose their splicing activity. The activity of the galectin-depleted extract can be reconstituted by the addition of either galectin-1 or galectin-3. These results suggest that galectins-1 and -3 serve as redundant splicing factors. Consistent with this notion, immunofluorescence staining showed that both galectins yielded a diffuse nucleoplasmic distribution, matching that of nascent transcripts and consistent with the hypothesis that bulk transcription and pre-mRNA processing occur throughout the nucleoplasm. Under some conditions, the galectins could be found in speckled structures and nuclear bodies but the prevailing thought is that these represent sites of storage and recycling rather than sites of action. Galectin-1 and galectin-3 bind directly to Gemin4, a component of the SMN core complex, which plays multiple roles in ribonucleoprotein assembly, including the biogenesis, delivery, and recycling of snRNPs to the spliceosome. Thus, galectin 1 and galectin-3 constitute a part of an interacting dynamic network of many factors involved in the splicing and transport of mRNA. PMID- 14758074 TI - Regulation of cellular homeostasis by galectins. AB - Members of the galectin family are presently known to participate in cellular homeostasis by modulating cell growth, controlling cell cycle progression, and inducing or inhibiting apoptosis. Both intracellular and extracellular activities of galectins have been described, with the former typically independent of lectin activity, and the latter mediated by lectin activity. Galectin-1 and -3 are recognized as activators and inducers of cell stasis in extracellular capacities. Galectin-1, -7, -8, -9 and -12 are characterized as promoters or inducers of apoptosis, while galectin-3 is demonstrated as an inhibitor of apoptosis intracellularly. Localization studies of galectins have established that these proteins can segregate into multiple intracellular compartments, and the preference for segregation is dependent on the status of the cell. Localization would, therefore, likely correspond to compartmental function. While galectin-1 and -3 have been the most abundantly expressed and extensively studied, and therefore, the members best understood, expanding interest in galectins has resulted in description of new members that display more restricted expression patterns, suggesting more specific activity. Nevertheless, as demonstrated for many members, it appears that a major feature of the galectin family is the homeostatic regulation of cells. PMID- 14758075 TI - Role of galectin-8 as a modulator of cell adhesion and cell growth. AB - Galectin-8 belongs to the family of tandem-repeat type galectins. It consists as several isoforms, each made of two domains of approximately 140 amino-acids, both having a carbohydrate recognition domain (CRD). These domains are joined by a 'link peptide' of variable length. The human galectin-8 gene covers 33 kbp of genomic DNA. It is localized on chromosome 1 (1q42.11) and contains 11 exons. The gene produces by alternative splicing 14 different transcripts, altogether encoding 6 proteins. Galectin-8, like other galectins, is a secreted protein. Upon secretion galectin-8 acts as a physiological modulator of cell adhesion. When immobilized, it functions as a matrix protein equipotent to fibronectin in promoting cell adhesion by ligation and clustering of a selective subset of cell surface integrin receptors. Complex formation between galectin-8 and integrins involves sugar-protein interactions and triggers integrin-mediated signaling cascades such as Tyr phosphorylation of FAK and paxillin. In contrast, when present in excess as a soluble ligand, galectin-8 (like fibronectin) forms a complex with integrins that negatively regulates cell adhesion. Such a mechanism allows local signals emitted by secreted galectin-8 to specify territories available for cell adhesion and migration. Due to its dual effects on the adhesive properties of cells and its association with fibronectin, galectin-8 might be considered as a novel type of a matricellular protein. Galectin-8 levels of expression positively correlate with certain human neoplasms, prostate cancer being the best example studied thus far. The overexpressed lectin might give these neoplasms some growth and metastasis related advantages due to its ability to modulate cell adhesion and cellular growth. Hence, galectin-8 may modulate cell-matrix interactions and regulate cellular functions in a variety of physiological and pathological conditions. PMID- 14758076 TI - Extracellular functions of galectin-3. AB - Galectin-3 has been suspected of modulating cell to extracellular matrix interactions in a novel fashion ever since it was first described. However, the rapid accumulation of research data in just the last 8 years alone has completely changed our perspective of this multifunctional protein. Its chimeric nature (consists of carbohydrate recognition and collagen like domains) somehow makes it suited to interact with a plethora of interesting extracellular matrix proteins some of which might enable it to cross the plasma membrane despite its lack of appropriate signal peptides. It is now becoming established as a mediator of signal transduction events on the cell surface as well as a mediator of a variety of extra-cellular processes such as kidney development, angiogenesis, neuronal functions, tumor metastasis, autoimmune disorders, endocytosis and possibly exocytosis. Nevertheless, it still retains its unique position as a mediator/modulator of cell to extracellular matrix adhesive interactions. Cells, particularly epithelial cells which lack galectin-3 expression, interact poorly with their extracellular matrices. In some of these processes, it functions as a matricellular protein, displaying both pro- and anti-adhesive properties. PMID- 14758077 TI - Expression of galectins in cancer: a critical review. AB - A large body of literature has examined and described galectin expression in cancer. Discrepancies have been observed in the reported data, which hampered clear understanding of the expression profiles. This relates to the use of different types of methods that evaluate either global or specific gene expression in heterogeneous cancer tissue samples, type of antibodies used in immunohistochemistry and procedures of comparison of gene expression. In this manuscript, we review the main data concerning expression of galectins in human cancer. Only galectin-1 and galectin-3, the most abundant and examined galectins, will be examined here. PMID- 14758078 TI - Galectin-3 and metastasis. AB - Galectin-3, a 31 kDa member of the beta-galactoside-binding proteins, is an intracellular and extracellular lectin which interacts with intracellular glycoproteins, cell surface molecules and extracellular matrix proteins. Galectin 3 is expressed widely in epithelial and immune cells and its expression is correlated with cancer aggressiveness and metastasis. Galectin-3 is involved in various biological phenomena including cell growth, adhesion, differentiation, angiogenesis and apoptosis. Recent research revealed that galectin-3 is associated with several steps of invasion and metastasis, like angiogenesis, cell matrix interaction, dissemination through blood flow and extravasation. Recently, we and others have shown that galectin-3 can be a reliable diagnostic marker in certain cancers and one of the target proteins of cancer treatment. In this review, we describe the involvement of galectin-3 in each steps of metastasis and clinical significance of galectin-3. PMID- 14758079 TI - 90K (Mac-2 BP) and galectins in tumor progression and metastasis. AB - Galectins and their ligands have been implicated in cell transformation and cancer metastasis, and found to have prognostic value. Mac-2 BP, also known as 90K, is a highly glycosylated, secreted protein extensively studied in human cancer, which binds galectin-1, galectin-3 and galectin-7. High expression levels of 90K are associated with a shorter survival, the occurrence of metastasis or a reduced response to chemotherapy in patients with different types of malignancy. The mechanisms underlying the prognostic significance of 90K and galectins in cancer are far from being understood, although they may be related to the ability of these proteins to interact and, to some extent, modulate cell-cell and cell matrix adhesion and apoptosis. The resulting scenario is even more complex, as data have been presented that all these proteins might be associated with either a positive or a negative outcome of the patients. It is hypothesised that different galectins and galectin ligands with overlapping or opposite functions, expressed in different tumors during the different steps of the metastatic cascade might play a crucial role in tumor progression. PMID- 14758080 TI - Human galectin-8 isoforms and cancer. AB - Galectins are animal lectins that can specifically bind beta-galactosides. Thirteen galectins have already been described. This review focuses on a specific member of this family: galectin-8. This galectin was discovered in prostate cancer cells eight years ago and has been studied extensively in the last few years. The galectin-8 gene ( LGALS8) encodes numerous mRNAs by alternate splicing and the presence of three unusual polyadenylation signals. These mRNAs encode six different isoforms of galectin-8: three belong to the tandem-repeat galectin group (with two CRDs linked by a hinge peptide) and three to the prototype group (with one CRD). Various studies showed that galectin-8 is widely expressed in tumor tissues as well as in normal tissues. The level of galectin-8 expression may correlate with the malignancy of human colon cancers and the degree of differentiation of lung squamous cell carcinomas and neuro-endocrine tumors. Recently, the differences in galectin-8 expression levels between normal and tumor tissues have been used as a guide for the selection of strategies for the prevention and treatment of lung squamous cell carcinoma. These experiments are still under investigation, but demonstrate the potential of galectin-8 research to enhance our understanding of, and possibly prevent, the process of neoplastic transformation. PMID- 14758081 TI - Shedding light on the immunomodulatory properties of galectins: novel regulators of innate and adaptive immune responses. AB - Galectins are a large family of structurally related beta-galactoside-binding proteins that play a pivotal role in the control of cell differentiation, proliferation, activation and apoptosis of many different cell types including immune cells. By crosslinking specific glycoconjugates, different members of the galectin family behave as pro-inflammatory or anti-inflammatory "cytokine-like" mediators, acting at different levels of innate and adaptive immune responses. Here we will review recent advances on the role of galectins in key events of the immune and inflammatory response, such as tolerance induction, cell cycle progression, cell adhesion, chemotaxis, antigen presentation and apoptosis. In particular we will examine the influence of individual members of the galectin family in the physiology of different immune cell types involved in innate and adaptive immune responses. Moreover, we will discuss the importance of these sugar-binding proteins as therapeutic targets in Th1- and Th2-mediated immune disorders, an exciting area for future research. PMID- 14758082 TI - Galectins as inflammatory mediators. AB - Over the last decade a vast amount of reports have shown that galectin-1 and galectin-3 are important mediators of inflammation. In this review we describe how the galectins may be involved in several parts of the inflammatory process, including the recruitment of neutrophils into an infected tissue and the recognition and killing of bacteria by activation of the tissue destructive phagocytic respiratory burst. During bacterial infection or aseptic inflammatory processes, galectins are produced and released by e.g. infected epithelium, activated tissue-resident macrophages and endothelial cells. These extracellular galectins may facilitate binding of neutrophils to the endothelium by cross linking carbohydrates on the respective cells. Further the galectins improve binding of the neutrophil to the extracellular matrix proteins laminin and fibronectin, and are potential chemotactic factors, inducing migration through the extracellular matrix towards the inflammatory focus. When the cells encounter bacteria, galectin-3 could function as an opsonin, cross-linking bacterial lipopolysaccharide or other carbohydrate-containing surface structures to phagocyte surface glycoconjugates. Both galectin-1 and galectin-3 have the capacity to induce a respiratory burst in neutrophils, provided that the cells have been primed by degranulation and receptor upregulation. The reactive oxygen species produced may be destructive to the invading micro-organisms as well as to the surrounding host tissue, pointing out the possible role of galectins, not only in defence toward infection, but also in inflammatory-induced tissue destruction. PMID- 14758083 TI - Seeing strangers or announcing "danger": galectin-3 in two models of innate immunity. AB - Recent investigations on the molecular mechanisms by which our immune system recognizes infections and initiates defense against those infections have led to the proposition of two models explaining the way our innate immunity system functions; the self-nonself model and the Danger model. In this review, the roles of galectin-3 in innate immunity against infections--host-pathogen interactions- will be discussed. We will shed light on the potential contribution of a beta galactoside binding mammalian lectin, galectin-3 as a molecule implicated in innate immunity from the angle of both the self-nonself model and the Danger model. PMID- 14758084 TI - Galectin-9 in physiological and pathological conditions. AB - We first cloned galectin-9 (Gal-9)/ecalectin as a T cell-derived eosinophil chemoattractant. Gal-9 plays a role in not only accumulation but also activation of eosinophils in experimental allergic models and human allergic patients, because Gal-9 induces eosinophil chemoattraction in vitro and in vivo and activates eosinophils in many aspects. Gal-9 requires divalent galactoside binding activity but not the linker peptide of Gal-9 to exhibit its biological functions, and an unidentified matrix metalloproteinase is involved in the release of Gal-9. Our recent studies also showed that Gal-9 has other functions, such as cell differentiation, aggregation, adhesion, and death. Now, we and other groups are on the way of investigating the regulation and function of Gal-9 in a variety of physiological and pathological conditions. In this article, we will show the possible role of Gal-9 in physiological and pathological conditions by using our recent findings. PMID- 14758085 TI - Galectins in parasite infection and allergic inflammation. AB - Galectins are increasingly recognised as important immunological mediators of homeostasis and disease regulation. This paper gives an overview of current knowledge of galectin involvement in parasite infection and allergic inflammation, two very different but immunologically linked phenomena. Galectins are produced by both the parasite and the host and appear to be intimately involved in parasite establishment, as well as directing the course of infection and the immune response. Host galectins have also been shown to be active participants in the recruitment of cells to sites of inflammation and modulating the effector function of mast cells, neutrophils and eosinophils. Moreover, the ability of galectins to induce differential expression of cytokine genes in leukocytes suggests that they are able to direct the nature of an adaptive immune response, in particular towards a T2-type allergic response. PMID- 14758086 TI - Insect galectins: roles in immunity and development. AB - As evidenced by the reviews in this special issue of Glycoconjugate Journal, much research is focused on determining functions for mammalian galectins. However, the identification of precise functions for mammalian galectins may be complicated by redundancy in tissue expression and in target cell recognition of the many mammalian galectins. Therefore, lower organisms may be useful in deciphering precise functions for galectins. Unfortunately, some genetically manipulable model systems such as Caenorhabditis elegans may have more galectins than mammals. Recently, galectins were identified in two well-studied insect systems, Drosophila melanogaster and Anopheles gambiae. In addition to the powerful genetic manipulation available in these insect models, there is a sophisticated understanding of many biological processes in these organisms that can be directly compared and applied to mammalian systems. Understanding the roles of galectins in insects may provide insight into precise functions of galectins in mammals. PMID- 14758087 TI - The involvement of galectin-1 in skeletal muscle determination, differentiation and regeneration. AB - The dogma that a cell is rigidly committed to one tissue type has been heavily challenged over the past few years with numerous reports of transdifferentiation of cells between different lineages. Cells capable of entering lineages other than that of their tissue of origin have been identified in several diverse tissues. Recently we have focussed on a non-committed myogenic cell within the dermis that is capable, under certain conditions, of expressing muscle specific markers and even fusing to the terminally differentiated stage of muscle cell development. We have identified galectin-1 as being a potent factor implicated in this process. In this review we discuss our findings and consider the involvement of galectin-1 in muscle determination, differentiation and regeneration. PMID- 14758089 TI - Inhibition of breast cancer regrowth and pulmonary metastasis in nude mice by anti-gastric ulcer agent, irsogladine. AB - Irsogladine is a commonly used anti-gastric ulcer agent in Japan, and recent in vivo studies have shown it to have anti-angiogenic properties. The exact role of irsogladine as an inhibitor of angiogenesis remains uncertain. In this study, we show that irsogladine inhibited breast cancer regrowth and pulmonary metastasis but had no anti-angiogenic function against HUVEC cells. Irsogladine failed to inhibit proliferation, tubular formation, and the uPA/MMP-1 mRNA expression of HUVEC cells. We also examined the effect of irsogladine in an orthotopic transplant model of human breast cancer metastasis in athymic mice. Human MDA-MB 435 cells were injected into the mammary fat pads. After 9 weeks, the tumors were resected under general anesthesia. Irsogladine or vehicle was given p.o. daily thereafter. Daily administration of irsogladine at 120 mg/kg per day over a 5 week period had no effect on the body weight of the mice. Tumor regrowth, average volume of pulmonary metastases, and the number of metastases were inhibited by 40, 48 and 64%, respectively. These results suggest that irsogladine may be useful in the breast cancer adjuvant setting. PMID- 14758088 TI - Galectins in kidney development. AB - Galectins are a family of proteins with overlapping but distinct carbohydrate binding specificities. They differ in cell-type and tissue distribution, and have various functions. Extracellularly several galectins can modulate cellular adhesive interactions and signalling pathways, effects that may be important in the establishment and maintenance of tissue organization during normal development. This review will summarise recent progress in defining the roles of galectins that are expressed in the kidney in normal development, and discuss the evidence linking aberrant expression of galectins with kidney disease. PMID- 14758090 TI - Energy balance in early breast cancer patients receiving adjuvant chemotherapy. AB - Weight gain is a common problem amongst women receiving adjuvant chemotherapy for early breast cancer. We undertook a study to determine the causes of this weight gain. Prospective measurements of body mass and composition (skinfolds, bioelectrical impedance, total body potassium), energy balance (resting energy expenditure dietary intake, and physical activity), were determined in 17 women during and in the 6 months after commencing adjuvant chemotherapy. Women gained significant amounts of weight (5.0 +/- 3.8; p < 0.01) and body fat (7.1 kg +/- 4.5; p < 0.01) over the year. Waist circumference (5.1 +/- 4.5 cm; p < 0.01) and abdominal skinfold (16.2 +/- 10 mm; p < 0.01) were also increased but there was a decline in fat free mass (FFM); 1.7 +/- 2.5 kg. Women due to receive adjuvant chemotherapy had a greater resting energy expenditure (REE) compared with healthy subjects (n = 21); 100.5 +/- 8.0% Harris Benedict compared to 94.5 +/- 8.4% Harris Benedict (p = 0.05). REE declined by 3% during adjuvant chemotherapy (p < 0.05), and remained depressed until at least 3 months posttreatment. There were no significant changes in dietary intake or physical activity over the year. Failure of women to reduce their energy intake to compensate for the decreased energy requirement may account for some of the weight gain. Treatment of adjuvant chemotherapy causes gain of body fat because of reduced energy expenditure, and the failure of women to reduce their energy intake to compensate for the decline in energy requirement during and in the 6 months posttreatment. Since weight gain impacts on survival, patients should be counselled to reduce energy intake and exercise during and after adjuvant treatment. PMID- 14758091 TI - Effect of progesterone receptor a predominance on breast cancer cell migration into bone marrow fibroblasts. AB - Women exposed to exogenous progesterone have increased breast cancer risk, but the mechanisms of progesterone involvement in breast cancer development are unknown. In human breast and endometrium, progesterone receptor (PR) isoform expression is disrupted in premalignant lesions and predominance of one isoform, usually PRA, in invasive cancers is associated with poorer prognosis. Disrupted PR isoform expression results in disrupted progestin regulation of cell morphology, including rounded morphology and decreased adherence of cells to tissue culture flasks. The purpose of this study was to test the hypothesis that predominance of PRA affects the interaction of breast cancer cells with a physiologically relevant stromal tissue, bone marrow stroma. T-47D breast cancer cells demonstrated the ability to migrate into bone marrow fibroblasts and this was inhibited by progestin treatment. The antiprogestin RU38486 abrogated the progestin effect on migration, demonstrating that it was PR-mediated. In cells expressing a predominance of PRA, after induction of a stably integrated inducible PRA construct, the ability of progestin to inhibit breast cancer cell migration was lost. A number of integrins were progestin regulated in T-47D cells, but there was no difference in the progestin effect in cells with PRA predominance, nor were the levels of focal adhesion proteins altered in these cells. This suggested that the lack of inhibition by progestin of breast cancer cell migration in cells with PRA predominance was not mediated by PRA effects on the membrane components of the adherens junctions. In summary, this study has shown that PRA predominance has a striking functional effect on breast cancer cell migration into stromal layers. PRA predominance may render breast cancer cells relatively resistant to the inhibitory effects of progestins and one consequence of this may be increased invasion of stroma. If borne out in vivo, these findings suggest that tumours with PRA predominance may be predisposed to cancer progression and this may signal a poorer prognosis in patients. PMID- 14758092 TI - Growth inhibitory activity of extracts and purified components of black cohosh on human breast cancer cells. AB - The purpose of this study was to determine whether black cohosh contains constituents that inhibit the growth of human breast cancer cells, and therefore might eventually be useful in the prevention or treatment of breast cancer. Black cohosh rhizomes were extracted with methanol/water and fractionated by solvent solvent partitioning to yield three fractions: hexane, ethyl acetate and water. The ethyl acetate fraction displayed the highest potency in two cell-based assays, growth inhibition and cell cycle analysis. This fraction inhibited growth of both the ER+ MCF7 and ER-MDA-MB-453 human breast cancer cell lines with IC50 values of about 20 and 10 micro g/ml, respectively. It also induced cell cycle arrest at G1 when tested at 30 micro g/ml and at G2/M at 60 micro g/ml in MCF7 cells. This suggests that the extract contains a mixture of components with the more active (or more abundant) causing G1 arrest and the less active causing G2/M arrest. We then examined specific components in this extract. The triterpene glycoside fraction obtained by polyamide column chromatography, and the specific triterpene glycosides actein, 23-epi-26-deoxyactein and cimiracemoside A, inhibited growth of the MCF7 human breast cancer cells and induced cell cycle arrest at G1. The most potent compound, actein, decreased the level of cyclin D1, cdk4 and the hyperphosphorylated form of the pRb protein and increased the level of p21cip1 in MCF7 cells, changes that may contribute to the arrest in G1. Further studies are in progress to identify the mechanisms by which actein and related compounds present in black cohosh inhibit growth of human breast cancer cells. PMID- 14758093 TI - Establishment of two hormone-responsive mouse mammary carcinoma cell lines derived from a metastatic mammary tumor. AB - We report the establishment of two mouse mammary cancer cell lines, MC7-2A and MC7-2B obtained from a mouse mammary carcinoma induced by medroxyprogesterone acetate (MPA) and maintained by syngeneic transplantation in BALB/c mice. They are epithelial (express cytokeratins) and express both estrogen receptors alpha (ERalpha) and progesterone receptors (PRs) isoforms A and B (western blots). In vitro, MPA inhibited 3H-thymidine uptake, starting from concentrations as low as 10(-13) M in MC7-2A and 10(10) M in MC7-2B; the antiprogestin RU 486 exerted a stimulatory effect at 10(-14) M in both cell lines; 17-beta-estradiol (E2) also exerted a stimulatory effect starting at 10(-10) M in MC7-2A and at 10(-13) M in MC7-2B. When transplanted in syngeneic mice, both cell lines originated adenocarcinomas that gave rise to lung metastases within 3 months. In in vivo studies, in MC7-2A, the antiprogestin inhibited completely tumor growth, E2 induced a slight although significant ( p < 0.05) stimulatory effect and MPA stimulated tumor growth while MC7-2B cells were unresponsive to all treatments. ER and PR were also expressed in tumors as assessed by immunohistochemistry. Two marker chromosomes were identified by FISH as translocations between chromosomes 4 and 7, and between chromosomes X and 2; the third marker chromosome remains unidentified. All these markers were also present in the parental tumor. A new marker, a centric fusion of chromosomes 2, was acquired in both cell lines. Considering that there are very few murine breast carcinoma responsive cell lines, these cells represent new tools in which the regulatory effect of hormones can be studied. PMID- 14758094 TI - Chemotherapy of nonelderly breast cancer patients by poverty-rate of area of residence in Connecticut. AB - For 684 nonelderly Connecticut women diagnosed in 1999 with early-stage breast cancer, routinely reported information on chemotherapy in a population-based cancer registry was supplemented by questionnaires to their physicians. Receipt of or recommendation for chemotherapy was associated with younger age, larger tumor size and positive lymph-node status, but not with higher poverty-rate (upper quintile) of census tract of residence at diagnosis. Similar studies are needed in other geographical areas that differ in socioeconomic indicators. PMID- 14758095 TI - Relevance of breast cancer cell lines as models for breast tumours: an update. AB - The number of available breast cancer cell (BCC) lines is small, and only a very few of them have been extensively studied. Whether they are representative of the tumours from which they originated remains a matter of debate. Whether their diversity mirrors the well-known inter-tumoural heterogeneity is another essential question. While numerous similarities have long been found between cell lines and tumours, recent technical advances, including the use of micro-arrays and comparative genetic analysis, have brought new data to the discussion. This paper presents most of the BCC lines that have been described in some detail to date. It evaluates the accuracy of the few of them widely used (MCF-7, T-47D, BT 474, SK-BR-3, MDA-MB-231, Hs578T) as tumour models. It is concluded that BCC lines are likely to reflect, to a large extent, the features of cancer cells in vivo. The importance of oestrogen receptor-alpha (gene ESR1 ) and Her-2/ neu ( ERBB2 ) as classifiers for cell lines and tumours is underlined. The recourse to a larger set of cell lines is suggested since the exact origin of some of the widely used lines remains ambiguous. Investigations on additional specific lines are expected to improve our knowledge of BCC and of the dialogue that these maintain with their surrounding normal cells in vivo. PMID- 14758096 TI - Cardiac resynchronization therapy: effects on exercise capacity in the patient with chronic heart failure. PMID- 14758097 TI - Optimizing risk stratification in cardiac rehabilitation with inclusion of a comorbidity index. AB - PURPOSE: The risk stratification criteria of the American Association of Cardiovascular and Pulmonary Rehabilitation include guidelines to be used in stratifying cardiac rehabilitation (CR) patients for risk of disease progression (long term) and clinical events (short term). Noncardiac comorbidities are not included as indicators in these criteria. This study was designed to ascertain the prevalence of noncardiac comorbidities among CR patients, and to assess their relation to the current risk stratification algorithm for clinical events. METHODS: Patients were stratified into high-, intermediate-, and low-risk groups according to the American Association of Cardiovascular and Pulmonary Rehabilitation risk stratification criteria for clinical events (ARSE) at program entry. Within each risk group, age, gender, race, and noncardiac comorbidities were ascertained. Comorbidities were summarized in a comorbidity index (CMI). The relation between clinical events and risk status by ARSE and CMI was evaluated by logistic regression. RESULTS: Among 490 patients (age, 60 +/- 12 years; 35% women; 30% nonwhite) enrolled in CR with ischemic heart disease, the number of comorbidities ranged from 0 to 7 (median, 2; 75th percentile, 3). The patients categorized in the three ARSE groups differed significantly in age and comorbidities. Although ARSE tended to identify patients with a greater comorbidity burden, 38% of the patients with a comorbidity index exceeding the 75th percentile were not classified in the highest ARSE group. Clinical events increased across ARSE and CMI risk strata. Both ARSE and CMI were independent predictors of events in an age-, gender-, and race-adjusted logistic regression analysis (ARSE odds ratio [OR], 1.56; 95% confidence interval [CI], 1.14-2.12; CMI OR, 1.23, 95% CI, 1.03a-1.47). Events were predicted best when both classifications were combined. Exploratory gender-specific analyses suggested that ARSE performed better among men than among women, whereas CMI was a more important predictor among women. CONCLUSIONS: To appreciate more fully the overall complexity of disease among CR patients, ARSE should be supplemented not only with the inclusion of cardiac risk factors, as suggested in the current guidelines, but also with an assessment of noncardiac comorbidities. PMID- 14758098 TI - The complexities of predicting risk in CVR patients. PMID- 14758099 TI - Social support as a predictor of participation in cardiac rehabilitation after coronary artery bypass graft surgery. AB - PURPOSE: Cardiac rehabilitation promotes recovery and enhances quality of life after a coronary artery bypass graft (CABG), but participation in such rehabilitation is low. The role of social support in promoting participation has been suggested by prior studies, but is not clearly defined. The purpose of this study was to investigate the role of social support as an independent predictor of participation in cardiac rehabilitation. METHODS: This study examined 944 patients who underwent first isolated CABG between May 1999 and February 2001, then were followed for 6 months after surgery. Social support before CABG and 6 weeks after CABG was assessed using the Enhancing Recovery in Coronary Heart Disease (ENRICHD) Social Support Inventory (ESSI) and evaluated for its association with participation in cardiac rehabilitation. RESULTS: Of 944 patients, 524 (56%) reported participation in rehabilitation. The participants were younger, better educated, more often employed, and less financially strained. The participants also had a lower prevalence of cardiovascular disease risk factors and better physical function. According to unadjusted analysis, the patients with low social support (ESSI 75 years vs. <65 years; OR, 0.40; 95% CI, 0.3-0.7) predicted decreased cardiac rehabilitation prescription. Conversely, previous history of dyslipidemia (OR,1.4; 95% CI, 1.04-1.8), post-myocardial infarction (OR, 2.8; 95% CI, 2.13-3.89), and a percutaneous intervention (OR,1.9; 95% CI, 1.3-2.7) predicted increased cardiac rehabilitation prescription. Severe left ventricular impairment (< or =35% vs >50%) was not an independent factor for cardiac rehabilitation prescription. At 6-month follow-up assessment, rehabilitation patients had a lower rate of hypertension (18% vs 27%), elevated low-density lipoprotein cholesterol (54% vs 62%), and continued smoking (34% vs 50%). CONCLUSIONS: The results of the PREVENIR survey underscore the low level of cardiac rehabilitation prescription in France, and the relative exclusion of women and elderly people. Among the risk factors, dyslipidemia and current smoking are more frequent among rehabilitated patients. These findings may help to modify the strategy for using cardiac rehabilitation after acute coronary syndrome, although it is an effective intervention for secondary prevention. PMID- 14758103 TI - Do men and women achieve similar benefits from cardiac rehabilitation? PMID- 14758105 TI - Hematology and oncology. PMID- 14758104 TI - Effects of pulmonary rehabilitation on dyspnea, quality of life, and healthcare costs in California. AB - PURPOSE: This study evaluated pulmonary rehabilitation as practiced in the general California medical community to determine its effectiveness in improving dyspnea and health-related quality of life and reducing the use of healthcare resources. METHODS: For this study, 10 established pulmonary rehabilitation programs agreed to collect common clinical health outcome data on consecutive patients over 2 years. The following three self-administered questionnaires were obtained before and after rehabilitation, then at 3-, 6-, 12-, and 18-month follow-up assessments: Medical Outcomes Survey Short Form (SF-36), University of California, San Diego Shortness of Breath Questionnaire (SOBQ), and Health Care Utilization in the preceding 3 months. Information also was collected on patient demographics, diagnostic categories, use of supplemental oxygen, and available spirometry and 6-minute walk tests. RESULTS: Nine centers enrolled 647 patients that met prespecified inclusion criteria. Of these, 521 completed the rehabilitation program and both the pre- and the postprogram assessment. At least two of the four follow-up assessments were completed by 415 patients in eight centers. The mean age of the patients was 68 years, and 42% were men. Overall, the forced expiratory volume in 1 second was 44% of the predicted value. There were few significant differences between the centers. The baseline outcome measures demonstrated marked symptoms, as evidenced by the mean SOBQ score (56.8) and the mean impaired quality of life results (SF-36 physical component score, 31.2; SF-36 mental component score, 47.5). These measures also showed high utilization of healthcare services over the preceding 3 months in terms of mean hospital stay (2.4 days), urgent care visits (0.4), physician visits (4.4), and telephone calls (2.7). After rehabilitation, there were significant improvements in symptoms and quality of life in all the centers, as evidence by mean changes of -6.8 for the SOBQ, 7.5 for the physical component score, and 3.9 for the mental component score). Over 18 months, benefits gradually declined, but levels remained above baseline values. There also were significant reductions in all measures of healthcare utilization. CONCLUSIONS: Pulmonary rehabilitation was effective in improving symptoms and quality of life and reducing the utilization of healthcare resources over 18 months. The results were consistent across participating centers despite variations in practice settings, patient referral patterns, and program structure. PMID- 14758107 TI - Congenital defects in the interferon-gamma/interleukin-12 pathway. AB - PURPOSE OF REVIEW: The aim of this review is to highlight the most recent and relevant advances in the interferon-gamma/interleukin-12 pathway, a pivotal player of the immune system, and their repercussions on basic and clinical aspects of science. RECENT FINDINGS: Newly described mutations are helping us to dissect the interferon-gamma/interleukin-12 pathway and its role in genetic infectious susceptibility and autoimmunity, and to reevaluate the pathophysiologic mechanisms involved in dominant and recessively inherited mutations. SUMMARY: The interferon-gamma/interleukin-12 pathway plays a central role in immune control of both environmental and autochthonous challenges, as reflected in human mutations and animal models. Besides being crucial for mycobacterial control, the interferon-gamma/interleukin-12 pathway is also involved in the pathogenesis of autoimmune disease, as well as tumor development and control. Genotype-phenotype correlations have been established for certain mutants in this pathway, some of which have therapeutic implications. PMID- 14758108 TI - Influence of race and socioeconomic status on outcome of children treated for childhood acute lymphoblastic leukemia. AB - PURPOSE OF REVIEW: Overall, childhood acute lymphoblastic leukemia is associated with an excellent outcome. The improvement in survival achieved during the last three decades is partially attributed to the identification of risk factors predicting a poor outcome and risk-stratified treatment of patients placed on well-designed therapeutic trials. Accordingly, it is important to continue to identify patient subgroups with differences in outcome to focus efforts to improve overall survival. Black children historically have been reported to have a poorer survival rate compared with whites, but limited information is available for children from other racial/ethnic backgrounds. RECENT FINDINGS: Several groups have published reports on ethnic and racial differences in survival after childhood acute lymphoblastic leukemia, with poorer outcomes reported for black children compared with whites reported by the majority of the studies. Limited information is available for children from other racial/ethnic backgrounds, such as Hispanics and Asians, but data indicate that Hispanics have poorer survival than whites, whereas Asians from the United States have outcomes that are as good or better than those of the whites, especially among the high-risk group treated with contemporary risk-based therapy. The influence of race and ethnicity on survival should be closely linked with socioeconomic status. However, few studies have specifically investigated the influence of nutrition and socioeconomic factors on the prognosis of children with acute lymphoblastic leukemia, and the results are conflicting. SUMMARY: Future studies need to focus on the reasons for these differences, including racial and ethnic differences in adherence with therapeutic protocols, and ethnic differences in drug metabolism and bioavailability of the agents commonly used in acute lymphoblastic leukemia, so that drug administration can be modified if needed. PMID- 14758109 TI - Inherited thrombocytopenias: toward a molecular understanding of disorders of platelet production. AB - PURPOSE OF REVIEW: To review the defined syndromes of inherited thrombocytopenia and discuss new genetic data for several disorders that shed light on the process of megakaryopoiesis. RECENT FINDINGS: The genes responsible for several inherited thrombocytopenias have been recently discovered, including congenital amegakaryocytic leukemia, amegakaryocytic thrombocytopenia with radio-ulnar synostosis, familial platelet syndrome with predisposition to acute myelogenous leukemia, Paris-Trousseau, Wiskott-Aldrich syndrome, and the May-Hegglin, Sebastian, Epstein, and Fechner syndromes. These clinical syndromes, combined with studies in mouse and in vitro models, reveal the importance of these genes for normal hematopoiesis. SUMMARY: Although inherited syndromes of thrombocytopenia are rare, characterization of mutations in these disorders has contributed greatly to our understanding of megakaryocyte and platelet development. A systematic registry of congenitally thrombocytopenic individuals would almost certainly lead to new genetic discoveries. PMID- 14758110 TI - Dyskeratosis congenita and telomerase. AB - PURPOSE OF REVIEW: Dyskeratosis congenita, a rare condition characterized by mucocutaneous abnormalities and bone marrow failure, is caused by inherited defects in the telomerase complex. Autosomal dominant dyskeratosis congenita is associated with mutations in the RNA component of telomerase, hTERC, while X linked dyskeratosis congenita is due to mutations in the gene encoding dyskerin, a protein implicated in both telomerase function and ribosomal RNA processing. This review highlights recent research on dyskeratosis congenita and its relevance to other fields, including cancer and aging. RECENT FINDINGS: Newly developed animal models suggest that defects in ribosomal RNA processing contribute to the phenotype of X-linked dyskeratosis congenita. Bone marrow dysfunction may be the first manifestation of dyskeratosis congenita in children, and hTERC mutations have been detected in a subset of patients presumed to have idiopathic aplastic anemia or myelodysplastic syndrome. In vitro studies suggest that hTERC mutations associated with dyskeratosis congenita or aplastic anemia either impair the specific activity of telomerase, decrease hTERC stability, or disrupt assembly of the telomerase complex. Recent clinical reports suggest that nonmyeloablative conditioning regimens afford better outcomes in patients with dyskeratosis congenita who require hematopoietic stem cell transplantation. SUMMARY: Studies of dyskeratosis congenita have shed light on the pathobiology of aplastic anemia and other forms of bone marrow dysfunction. It seems likely that mutations in other genes involved in telomere maintenance will be linked to bone marrow failure or other human diseases. Genetic testing for occult dyskeratosis congenita may be warranted in selected patients with aplastic anemia or myelodysplastic syndrome, as this may impact the choice of therapies. PMID- 14758111 TI - Traumatic patellar dislocation in children and adolescents: treatment update and literature review. AB - PURPOSE OF REVIEW: Traumatic patellar dislocation in children and adolescents is a complex problem resulting from a range of anatomic and mechanical conditions. A careful review of the literature demonstrates certain risk factors that predispose children to recurrent dislocation. With this understanding, these conditions can be managed more aggressively in the hope of an improved outcome. Additionally, a vast number of surgical procedures to correct patellar instability have been described, and recent cadaveric studies are now guiding surgical interventions. RECENT FINDINGS: Risk factors for recurrent dislocation may include various skeletal abnormalities, increased quadriceps angle, generalized ligamentous laxity, and family history. Recent anatomic and biomechanical studies have demonstrated that the medial patellofemoral ligament and the vastus medialis obliquus are the primary restraints to lateral translation and ultimately dislocation of the patella. Management should therefore be directed both at correcting anatomic abnormalities when indicated and at reconstruction of medial restraints to patellar tracking. SUMMARY: The recommendation for management of a traumatic patellar dislocation in a skeletally immature patient is initially conservative, emphasizing early motion and quadriceps strengthening. However, in patients for whom conservative management has failed or who are at particularly high risk for dislocation and require surgical intervention, repair or reconstruction of the medial patellofemoral ligament is the treatment of choice. Recent works have included investigation of less invasive techniques in children. PMID- 14758112 TI - Symptomatic spondylolysis: diagnosis and treatment. AB - PURPOSE OF REVIEW: Approximately 35% of adolescents experience back pain. In athletic adolescents, spondylolysis is the most common offending cause. With growing numbers of adolescents participating in sports with higher levels of intensity, spondylolysis is becoming an increasingly common clinical problem. RECENT FINDINGS: A recent report demonstrated the benign natural history of asymptomatic spondylolysis. However, long-term follow-up studies of patients who experience painful spondylolysis as adolescents remain unavailable. Modern imaging modalities have led to earlier diagnosis with greater accuracy. Conservative management with bracing continues to be a mainstay of treatment. In patients who are not helped by conservative therapy, recent studies have demonstrated the satisfactory long-term results of surgical repair. SUMMARY: The long-term sequelae of symptomatic spondylolysis and unhealed pars defects require investigation. MRI promises to be a valuable tool for diagnosis and clinical stratification, but further studies are necessary to demonstrate its clinical utility. PMID- 14758113 TI - Overuse injuries in pediatric athletes. AB - PURPOSE OF REVIEW: The number of children participating in sports has risen steadily over the past few decades. As a result, the incidence of pediatric overuse injuries is rapidly increasing. Currently, primary care physicians are at the frontline in treating these injuries. It is important for these clinicians to be familiar with these types of injuries and their treatment options. RECENT FINDINGS: This article reviews overuse injuries on the basis of location. It discusses the most recent literature describing their presentations, their treatment options, and suggested criteria for return to play. SUMMARY: In summary, because the number of these injuries is on the rise, it is important for the physician to be aware of the clinical manifestations of overuse injuries, to prescribe current recommended treatments, and to educate patients in proper athletic conditioning. PMID- 14758114 TI - Surgical treatment of pediatric femoral shaft fractures. AB - PURPOSE OF REVIEW: Femoral shaft fractures are among the most common fractures in children. Depending on the patient's age, fracture location, pattern, mechanism of injury, and associated injuries, several different treatment options exist. The purpose of this review is to discuss these different clinical situations and the recommended treatment methods, as well as to characterize the latest literature and recommendations. RECENT FINDINGS: In the past several years, there have been significant changes in the approach to the treatment of pediatric femoral shaft fractures, particularly in school-aged children. Young children have traditionally been treated conservatively with good results, and this method is still currently advocated. Adolescents over the age of 12 are generally treated with rigid intramedullary rods. However, in children between the ages of 5 and 12, new surgical treatment modalities have been tested with good outcomes, and, as new data emerge, these methods are becoming preferable to conservative treatment. SUMMARY: Children who sustain femoral shaft fractures can present difficult challenges to both orthopedists and pediatricians. A recent shift in treatment in children between ages of 5 and 12 from nonoperative to surgical intervention has led to shorter hospital stays and earlier return to activity with reliable fracture healing. PMID- 14758115 TI - Impact of pneumococcal and influenza vaccines on otitis media. AB - PURPOSE OF REVIEW: The pneumococcal conjugate vaccine and influenza virus vaccines have the potential to reduce the risk of acute otitis media. Several recent studies have evaluated the impact of each of these vaccines on the incidence of acute otitis media. RECENT FINDINGS: In controlled studies, pneumococcal conjugate vaccine has resulted in a reduction in the incidence of acute otitis media of approximately 6%. Inactivated influenza vaccine does not appear to reduce the incidence of acute otitis media in children 6-24 months of age, but a few studies, each with design or methodological limitations, suggest that it may reduce the incidence among older children. One study reported that the live-attenuated, cold-adapted trivalent influenza vaccine reduced the incidence of febrile otitis media by 30%. Additional studies are needed to clarify the value of influenza vaccines to prevent acute otitis media. SUMMARY: The pneumococcal conjugate vaccine and the influenza vaccines may have a limited beneficial impact on the incidence of acute otitis media. Nevertheless, at this time, these vaccines should be promoted because of their primary benefits, which are reduced serious pulmonary and invasive infections, and not because they may reduce the incidence of acute otitis media. PMID- 14758116 TI - Severe acute respiratory syndrome: public health response and clinical practice update for an emerging disease. AB - PURPOSE OF REVIEW: Severe acute respiratory syndrome is an emerging infectious disease with high morbidity and mortality and potential for global spread. This review highlights the salient points of the 2002-2003 outbreak to help clinicians with early recognition, treatment, disease control, and prevention. RECENT FINDINGS: Severe acute respiratory syndrome is a respiratory illness caused by a novel coronavirus, severe acute respiratory syndrome-coronavirus. The disease was first recognized in Asia in February 2003 and, over the next several months, spread to more than two dozen countries in North and South America, Europe, and Asia. The disease is characterized by fever with symptomatic and radiographic evidence of pneumonia. SUMMARY: Because diagnosis cannot be established by clinical findings alone, epidemiologic findings (including history of travel to affected areas, close contact with other case patients, or linkage to clusters of unexplained pneumonia) play a critical role in the diagnostic evaluation. Infection control procedures (including droplet and airborne precautions) are critical for preventing transmission. PMID- 14758117 TI - Hantavirus infection in children. AB - PURPOSE OF REVIEW: This article focuses on recent developments in knowledge about hantavirus infections and hantavirus cardiopulmonary syndrome in children. We highlight clinical characterization, epidemiology, pathogenesis, diagnostic techniques, and current alternatives for treatment and prevention. RECENT FINDINGS: After the first description of hantavirus pulmonary syndrome (HPS) in 1993 in the United States, new cases of HPS and new hantavirus species have been described throughout the Americas. The factors involved in the expression of hantavirus disease have, in part, been recognized, but there have been descriptions of newer viruses and newer rodent reservoirs. Several seroprevalence studies suggest that the virus-host interaction has been taking place for many years, and changes in human behavior and wild rodent ecology, sometimes secondary to industrial progress, facilitate the clinical recognition of disease. Sin nombre virus (SNV) and Andes virus (ANDV) are examples of the same disease with differences in the virus virulence and in the host response. The North American syndrome and the Southern HPS differ in epidemiologic patterns and in the spectrum of disease. SUMMARY: Currently, no Food and Drug Administration (FDA) approved antiviral drugs, vaccines, or immunotherapeutic agents are available for treatment of the disease, and therapy is primarily supportive. Intensive care medicine has played an outstanding role in decreasing the lethality of HPS. A ribavirin trial in the United States did not support the use of the drug in fully developed HCPS. Recently published data suggest that a strong neutralizing antibody response may be a predictor of effective clearance of and recovery from SNV infection. This has raised the possibility that passive immunotherapy may be useful in HCPS. Extensive work has been done to develop a hantavirus vaccine, but at present it seems unlikely that a vaccine will be in commercial development in the near future. PMID- 14758118 TI - Newer treatments for HIV in children. AB - PURPOSE OF REVIEW: Several new antiretroviral agents have been introduced into pediatric and adult use. This review will summarize information about these new agents and other recent advances in the care of HIV-infected children. RECENT FINDINGS: New drugs like tenofovir, emtricitabine, and enfuvirtide are being rapidly introduced into antiretroviral treatments for adult patients. In addition, some well-established drugs are being modified to make them more convenient (specifically didanosine and stavudine). Unfortunately, pediatric data lag for these new agents, in some cases because of complicated pharmacokinetics in children. At the same time, critical information on how to use established drugs like nelfinavir and efavirenz in younger children is slowly becoming available. Although antiretroviral treatment in children has often been initiated at standard doses of milligrams per kilogram, and susceptibility to drug was presumed in individuals without a previous history of exposure, recent data show that some primary infections are caused by drug-resistant virus, and there is a tremendous variability in serum drug levels in children. Researchers and clinicians should consider the role of baseline antiretroviral susceptibility testing and therapeutic drug monitoring to identify the optimal treatment for each child. SUMMARY: New therapeutic options for children with HIV infection are becoming available as the pharmacokinetics and best strategies for use of newer drugs are studied. PMID- 14758119 TI - Varicella zoster virus infections in children after the introduction of live attenuated varicella vaccine. AB - PURPOSE OF REVIEW: Varicella zoster virus is the cause of both varicella (chickenpox) and herpes zoster (shingles). A live attenuated varicella vaccine was developed in 1974 and was approved in 1995 by the United States Food and Drug Administration for administration to both healthy children (>12 months of age) and adults who are susceptible to varicella. Many studies have shown varicella vaccine to be highly effective. Widespread use of the varicella vaccine in the United States has led to important changes in the epidemiology of the infection. The purpose of this review is to summarize the most recent and important findings regarding the impact that the widespread use of varicella vaccine has had on the epidemiology of varicella zoster infections in children. RECENT FINDINGS: Data from the United States Centers for Disease Control and Prevention have shown a dramatic decline in the incidence of varicella (76 to 87% from 1995 to 2000), with the greatest decline observed in preschool children, as well as a reduction in the number of hospitalizations for cases of varicella. However, as the proportion of children in the United States who have received the varicella vaccine has increased there have been several recent reports in which the effectiveness of the vaccine was substantially lower than expected. In particular, reports during outbreaks of varicella in children have noted increases in breakthrough disease in children who were vaccinated before the age of 15 months, in children with asthma, in those who received the varicella vaccine soon after the measles, mumps, and rubella vaccine (<28 days), and in children who received the vaccine more than 3 years before the development of disease. SUMMARY: Although reports of outbreaks of chickenpox in highly immunized groups have raised questions regarding the need for changes to the current vaccination policy, data undeniably indicate that immunization with varicella vaccine has been and continues to be successful in reducing the burden of disease in children and that universal immunization should continue to be a priority in the United States PMID- 14758120 TI - Urinary tract infections in infants and children: a comprehensive overview. AB - PURPOSE OF REVIEW: Urinary tract infections are the most common serious bacterial infections in infants and young children. This review focuses on new additions to the literature for the period August 22, 2002, to August 21, 2003. RECENT FINDINGS: There is still considerable interest in determining which test is best to predict the likelihood of a positive urine culture in children at risk for urinary tract infection. One new analysis and several older analyses suggest that the finding of pyuria, as measured by at least 10 leukocytes/mm3 on unspun urine is a very valuable cutoff for identifying infants for whom urine culture is warranted. Several new investigations have studied the value of various imaging studies in children with urinary tract infections. It has been shown that the finding of vesicoureteral reflux is variable and that single studies may underestimate or overestimate the degree of reflux. The natural history for lower grades of reflux (grades 1, 2, and 3) is spontaneous resolution at a rate of 13% per year. The rationale for the determination of the degree of reflux by voiding cystourethrogram is to guide the institution of antimicrobial prophylaxis or surgical intervention until the reflux resolves. This is based on the assumption, as yet unproven, that these interventions will prevent or decrease reinfection and thereby prevent the development of renal scarring. Data are presented indicating that there is still no evidence that this assumption is correct. SUMMARY: Continued attention to the need for and benefit of imaging procedures in children with urinary tract infection mandates that there be a randomized, controlled prospective trial of antimicrobial prophylaxis versus no treatment for children with various degrees of reflux. PMID- 14758121 TI - The changing picture of hepatitis A in the United States. AB - PURPOSE OF REVIEW: Hepatitis A causes approximately half of the cases of viral hepatitis in the United States. Since 1999, routine hepatitis A immunization of children in areas of the United States with high rates of hepatitis A has been recommended. RECENT FINDINGS: There has been an increasing appreciation of the role of young children with asymptomatic or inapparent infection as the community reservoir of hepatitis A virus. Epidemiologic studies have demonstrated striking geographic variations in the incidence of hepatitis A in the United States. On the basis of this understanding, recommendations for control of hepatitis A were updated in 1999 to include routine vaccination of children living in states, counties, and communities with high rates of hepatitis A. Routine hepatitis A vaccination of children in areas with high rates of hepatitis A is a cost effective strategy to reduce the incidence of hepatitis A. SUMMARY: Improved understanding of the epidemiology and transmission of hepatitis A combined with the availability of effective hepatitis A vaccines have dramatically reduced the burden of hepatitis A in the United States. PMID- 14758122 TI - Fever without apparent source on clinical examination. AB - PURPOSE OF REVIEW: Episodes of acute illness associated with fever are common in children. Less common but also diagnostically challenging are episodes of prolonged fever in children. This article reports recent literature that has focused on the epidemiology, pathophysiology, clinical and laboratory evaluation, and treatment of both types of episodes. RECENT FINDINGS: A number of articles reviewed the epidemiology in several countries of bacteremia in children, including bacteremia caused by Streptococcus pneumoniae and Salmonella sp. Host susceptibility factors for bacterial illness, such as malnutrition, were reported. Diagnostic issues concerning bacteremia, including the technique of venipuncture and urine antigen testing, were of interest. A major study was published that reported a preventive approach through immunization to one type of bacteremia that caused by S. pneumoniae. Other studies addressed the issues of choice of therapy and duration of therapy in selected bacteremic disease, such as dental bacteremias and bacteremia, that were caused by Brucella melitensis. Lastly, several causes of prolonged fever (such as hemophagocytic syndrome and atypical cat scratch disease) were reported. SUMMARY: During the review period, studies addressed diverse issues related to acute and prolonged episodes of fever in children. There was a particular emphasis on the epidemiology, diagnosis, prevention, and treatment of bacteremic illness in children. PMID- 14758123 TI - Current challenges in lower respiratory infections in children. AB - PURPOSE OF REVIEW: Lower respiratory infections threaten the health of children worldwide. Streptococcus pneumoniae remains the most common bacterial cause of lower respiratory infection in children, whereas viral pathogens dominate as a more common cause of lower respiratory infection illness in infants and children overall. The diagnosis and clinical management of lower respiratory infections pose challenges to pediatric health providers as new technology is developed and new pathogens emerge in the spectrum of clinical disease. RECENT FINDINGS: Human metapneumovirus is now recognized as a cause of lower respiratory infection disease in children, and coronavirus has been linked to epidemics of severe acute respiratory syndrome. Respiratory syncytial virus continues to be a major source of viral lower respiratory infection illness in children and can lead to childhood asthma. Treatment for respiratory syncytial virus bronchiolitis depends largely on the severity of disease and the course of clinical symptoms. The diagnosis of bacterial lower respiratory infection disease remains a clinical challenge, but new methods to detect S. pneumoniae, or Chlamydia pneumoniae and Mycoplasma pneumoniae may facilitate the clinical management of these illnesses. As immunization against S. pneumoniae becomes more widely used, the complications of bacterial lower respiratory infections will diminish markedly. SUMMARY: Future progress in the clinical management of lower respiratory infection diseases will entail improved methods of early diagnosis, broader options for treatment, and better defined clinical parameters for triage and follow-up of children with lower respiratory infections. PMID- 14758125 TI - A patient questionnaire for radiation-induced brachial plexopathy. AB - We analyzed the usefulness of a symptom questionnaire to screen for radiation induced brachial plexopathy (RIBP) after breast cancer treatment. Four questions addressed distal and proximal paresis: impaired hand functions, problems raising the arm, carrying weights, and lifting objects from a high shelf. Eighty-one relapse-free patients were neurologically examined. Treatment was mastectomy (51%) or breast-conserving surgery (49%), radiotherapy to the supraclavicular +/- axilla with median 60 Gy maximum dose. Sixty-five subsequent control patients had breast-conserving surgery and radiotherapy to the breast only with 55 Gy median dose. Median follow up was 10 and 7.4 years, respectively. Sixteen patients had RIBP, 7 had Radiation Therapy Oncology Group (RTOG) grade 1, 4 grade 2, 3 grade 3, and 2 grade 4 RIBP. Thirty-seven patients had fibrosis and 32 had arm edema. Four patients with RIBP had no fibrosis (n = 2) or fibrosis of the axilla only (n = 2). Specificity of the question "impaired hand functions" for RIBP was 0.66 (95% confidence interval [CI], 0.51-0.78); sensitivity was 0.80 (95% CI, 0.52 0.96). Specificity of the question "raising the arm" was 0.98 (95% CI, 0.9-0.99) and sensitivity was 0.18 (95% CI, 0.04-0.45); the rate of false-positive control patients was 3%. In multivariate analysis, "impaired hand functions" and fibrosis were independent indicators of RIBP (P <0.005). Patients with breast irradiation only stated moderate/pronounced impaired hand functions; and problems carrying weights and lifting objects from a high shelf in 38%, 58%, and 77%, not significantly different from patients with RIBP or the patients with supraclavicular radiation. RIBP is not necessarily associated with fibrosis. The aim of the questionnaire was screening of a population at risk for RIBP. In this group, the question "problems raising the arm" detected severe RIBP with high specificity. Negation of "impaired hand functions" excludes RIBP. Both questions should be included in follow-up questionnaires. PMID- 14758126 TI - Ki-67 proliferation index: correlation with prognostic parameters and outcome in multiple myeloma. AB - The nuclear protein Ki-67 is a proliferation index, as it is expressed only by dividing cells. In this study, we investigated the clinical significance of Ki-67 determination on bone marrow biopsies of 35 patients with newly diagnosed multiple myeloma (MM). We examined the correlation of Ki-67 with other MM proliferation-related factors: interleukin-6 (IL-6), IL-10, bone marrow infiltration by plasma cells, serum lactate dehydrogenase (LDH), and beta 2 microglobulin (b2M). Ki-67 expression was also correlated with the survival rate of the patients. The results showed that Ki-67 expression increases with increasing stage of disease according to Durie-Salmon (classification stage III vs. I and II, p < 0.001). Furthermore, infiltration, IL-6, LDH, and b2M increase significantly with advancing stage of disease (p < 0.004). All parameters studied were significantly higher in patients versus controls. Ki-67 correlated with IL-6 (r: 0.422, p < 0.01), LDH (r: 0.437, p < 0.01), and b2M (r: 0.478, p < 0.004). There was a marked difference in survival between patients with MM with Ki-67 greater than 8% and patients with Ki-67 less than 8%, in favor of the latter (p < 0.07). We conclude that Ki-67 determination during routine pathological analysis of bone marrow in newly diagnosed MM could provide useful information about the proliferative activity and prognosis of the disease. PMID- 14758127 TI - Docetaxel versus paclitaxel for adjuvant treatment of ovarian cancer: case control analysis of toxicity. AB - The objective of this study was to evaluate the toxicity profile of docetaxel/carboplatin versus paclitaxel/carboplatin. All patients with primary ovarian, fallopian tube, or peritoneal malignancies treated with docetaxel and platinum at the University of Iowa between January 1996 and June 1999 were identified. Controls, treated with paclitaxel and platinum, were matched for age, date of diagnosis, type of cancer, stage, and residual disease. Toxicity was evaluated prior to each cycle and was graded according to the Gynecologic Oncology Group criteria. Twenty patients were identified in each group and evaluated. In the docetaxel/carboplatin group, sixteen (80%) patients experienced hematologic toxicity. Nine (45%) had grade III or IV neutropenia and fever developed in two of these patients. Grade III or IV thrombocytopenia developed in two patients. In contrast, among the paclitaxel/carboplatin group, grade III or IV neutropenia developed in only three patients (p < 0.05) and grade III or IV thrombocytopenia developed in two patients. There were no significant differences between the two groups with regard to gastrointestinal or renal toxicity. In the paclitaxel/carboplatin group, 13 patients developed neuropathy compared to only 2 patients (10%) in the docetaxel/carboplatin group (p < 0.05). There was no difference in the clinical response between the two treatment groups. In conclusion, neutropenia was more common with the docetaxel/carboplatin regimen, whereas neuropathy was more common in the paclitaxel-based regimen. The therapeutic efficacy was equivalent between the two groups. PMID- 14758128 TI - Phase II trial of vinorelbine plus doxorubicin in relapsed small-cell lung cancer: CALGB 9332. AB - Small-cell lung cancer that progresses after initial response may still be sensitive to systemic treatment. This study assessed doxorubicin plus vinorelbine tartrate (Navelbine Injection) in patients who had no prior exposure to these agents. Treatment consisted of vinorelbine at 25 mg/m2 on days 1 and 8 and doxorubicin at 50 mg/m2 on day 1 of each 21-day cycle. The trial was stopped early because of excessive toxicity. The partial response rate was 26.7%. Toxicities included grade IV neutropenia in 73%, and febrile neutropenia and/or sepsis in 60%. Three patients died from sepsis during cycle 1. Performance status 2 was significantly associated with febrile neutropenia (p = 0.044). Although this regimen had some activity, the toxicity precluded further evaluation. PMID- 14758129 TI - Effects of carbon-ion beams on human pancreatic cancer cell lines that differ in genetic status. AB - The relative biologic effectiveness (RBE) of carbon-ion beams at 3 different linear energy transfer (LET) values (13, 50, and 80 keV/microm) accelerated by the Heavy Ion Medical Accelerator in Chiba on human pancreatic cancer cell lines differing in genetic status was determined. The RBE values were calculated as D10, the dose (Gy) required to reduce the surviving fraction to 10%, relative to X-rays. We also investigated apoptosis and the relationship between D10 and the cell cycle checkpoint using morphologic examination and flow cytometry analysis, respectively. The RBE values calculated by the D10 values ranged from 1.16 to 1.77 for the 13-keV/microm beam and from 1.83 to 2.46 for the 80-keV/microm beam. A correlation between the D10 values of each cell line and intensity of G2/M arrest was observed. In contrast, LET values did not clearly correlate with induction of apoptosis. These results suggest that carbon-ion beam therapy is a promising modality. Elucidation of the mechanisms of G2/M arrest and apoptosis may provide clues to enhancing the effects of radiation on pancreatic cancer. PMID- 14758130 TI - Sinonasal teratocarcinosarcoma. AB - Sinonasal teratocarcinosarcoma (SNTCS) is a distinctly rare tumor characterized by a variegated histologic architecture of epithelial and mesenchymal components. By reported accounts, SNTCS is a highly malignant tumor displaying rapid, aggressive growth. Prognosis is poor: less than 45% of all patients survive past 5 years. Combination surgery and radiotherapy currently appear to be the most effective treatment. This report presents a 76-year-old African American man with a SNTCS in the right nasal cavity and paranasal sinuses. The patient was treated with combination surgical excision and postoperative radiation therapy. The clinical and pathologic features and clinical course will be discussed. PMID- 14758131 TI - Temozolomide in combination with irinotecan for treatment of recurrent malignant glioma. AB - The prognosis for patients with recurrent malignant glioma is poor. Both temozolomide and irinotecan have been shown to be active in this disease. A study was performed combining temozolomide 200 mg/m2 daily for 5 days and irinotecan 125 mg/m2 on days 6, 13, and 20 initially (Schedule A) and then changed to (Schedule B) temozolomide 200 mg/m2 daily for 5 days and irinotecan 350 mg/m2 on day 6. Each cycle was 28 days. All patients with recurrent tumor had to complete two cycles of therapy to be evaluable. Six cycles of treatment were provided for all responders. Thirty-two patients were treated, 6 with schedule A, 24 with schedule B, and 2 initially schedule A and then switched to schedule B. Eighteen patients (56%) had glioblastoma and 14 patients and anaplastic glioma (AOA 8, anaplastic astrocytoma 4, AO 2). Eighty-three percent (15/18) of patients with glioblastoma responded (complete response [CR] 2, partial response [PR] 3, stable disease [SD] 10). Median duration of response was 24 weeks, and 6-month progression-free survival (PFS) was 39% (7/18). Fourteen patients with anaplastic glioma were treated and all responded (CR 3, PR 2, SD 9). Median duration of response was 29 weeks and 6-month PFS was 71% (10/14). Grade IV leukopenia occurred in one patient and grade IV thrombocytopenia in two patients. Two patients were admitted to the hospital for neutropenic fever. Nonhematologic toxicity was mild and mostly gastrointestinal. These results demonstrate a favorable response and low toxicity with combined irinotecan and temozolomide therapy and warrant further clinical evaluation. PMID- 14758133 TI - Paclitaxel and carboplatin as second-line therapy in women with platinum sensitive ovarian carcinoma treated with platinum and paclitaxel as first-line therapy. AB - The study was performed to assess response rate, progression-free interval (PFI), and side effects of the combination paclitaxel and carboplatin as second-line therapy among women with platinum-sensitive epithelial ovarian carcinoma (EOC). Thirty women who achieved partial surgical response at second-look surgery (n = 8) or who had recurrence (n = 22) more than 6 months after treatment with platinum-based chemotherapy were treated with paclitaxel (135 mg/m2 for 3 hours) and carboplatin (area under the concentration-time curve 5) every 3 weeks. Response rate, PFI, and side effects of treatment were recorded. One hundred sixty-seven cycles of treatment (median = 6, range = 2-11) were administered. Among 22 patients with measurable or assessable disease, 14 had complete response and 3 had partial response. Five patients had progressive disease. The overall response rate was 77%. The median PFI was 10 months (range = 1-29). Among 22 patients in whom recurrence or progression developed after second-line therapy, the median interval was 9 months (range = 1-26). The incidence of grade III or IV neutropenia, leukopenia, and thrombocytopenia was 48%, 27%, and 3%, respectively. One patient discontinued treatment secondary to persistent thrombocytopenia. Eight patients died secondary to their disease. It was concluded that the combination paclitaxel and carboplatin has a high success rate, long duration of response, and is well tolerated as a second-line therapy among patients with platinum-sensitive EOC. PMID- 14758132 TI - Simultaneous mutations in K-ras and TP53 are indicative of poor prognosis in sporadic colorectal cancer. AB - Despite the fact that the mutations in K-ras codon 12 and TP53 are common abnormalities in colorectal cancer, the determination of K-ras mutation combined with TP53 gene mutation, with diagnostic and prognostic purposes is still controversial. We have analyzed K-ras and TP53 mutations in 77 sporadic colorectal adenocarcinomas by means of polymerase chain reaction and sequencing. We observed a negative correlation between both K-ras and TP53 mutations. Patients with mutations in K-ras but not in TP53 exhibited worse survival rates than those with mutations in TP53 and not in K-ras. Moreover, we found the worst outcome in patients with mutations in both K-ras and TP53. These results may relate to the previously published data about primary human and rodent cells, in which transformation by Ras require either a cooperating oncogene or the inactivation of tumor suppressors such as p53 or p16. In conclusion, simultaneous mutations in K-ras and TP53 are indicative of a worse prognosis in sporadic colorectal cancer. PMID- 14758134 TI - Phase II study of external irradiation and weekly paclitaxel for nonmetastatic, unresectable pancreatic cancer: RTOG-98-12. AB - Unresectable cancer of the pancreas was treated with the combination of weekly paclitaxel and external beam irradiation in an effort to improve palliation and extend life expectancy. One hundred twenty-two patients were entered in a multicentered protocol. Thirteen patients were either ineligible, cancelled, or had delinquent data, thus providing 109 for analysis. Unresectable cancer was based on imaging studies (computed tomography or magnetic resonance imaging), all had histologic proof of adenocarcinoma, and none had evidence of metastatic disease or peritoneal seeding. Image-guided radiotherapy treatment consisted of 50.4 Gy in 28 fractions over 5.5 weeks with coplanar anterior/posterior and lateral ports. An initial dose of 45 Gy was given to fields covering the primary tumor plus the regional peripancreatic, celiac, and porta hepatis lymph nodes. A cone down field was used for the last three fractions to encompass the gross tumor volume with a 1- to 1.5-cm margin. Paclitaxel was administered weekly with irradiation in a dosage of 50 mg/m2 as a 3-hour infusion. The median age was 63 and 53% were female. The Karnofsky performance status was greater than or equal to 80 in 81%. Eighty percent were classified T3 or 4; 20% had N1 disease. The primary tumor was located in the pancreatic head in 65%. Eighty-five percent received all six cycles of paclitaxel per protocol, whereas 93% received irradiation with acceptable protocol variation. Field placement, total dose, fractionation, and overall treatment time were given per protocol in greater than or equal to 90%. Acute toxicity (worst per patient) occurred in 39% with grade III (35% of these were asymptomatic neutropenia), 5% with grade IV, and one patient died of infection during the fourth cycle of chemotherapy (grade V). The median follow-up time for alive patients is 20.6 months (range 5-30). The median survival is 11.2 months (95% CI 10.1, 12.3) with estimated 1- and 2-year survivals of 43% and 13%, respectively. External irradiation plus concurrent weekly paclitaxel is well tolerated when given with large-field radiotherapy. The median survival is better than historical results achieved with irradiation and fluoropyrimidines. These data provide the basis for a new Radiation Therapy Oncology Group trial using paclitaxel and irradiation combined with a second radiation sensitizer, gemcitabine, now under way. PMID- 14758136 TI - Multimodality therapy for thymic carcinoma (TCA): results of a 30-year single institution experience. AB - SUMMARY: The aim of this study was to correlate the clinicopathologic features and therapeutic approaches with the outcome of patients with thymic carcinoma (TCA), an aggressive, uncommon malignancy of the anterior mediastinum. TCA is morphologically distinct from thymoma, a cytologically bland, often encapsulated, locally invasive, rarely metastatic tumor. The Roswell Park Cancer Institute tumor registry was used to identify patients with TCA or invasive thymic neoplasm of the epithelial type (TNET). Between 1971 and 2001, 22 patients had a pathologic diagnosis of TCA and/or TNET. The mean age at diagnosis was 53 years (range: 19-77), and the male/female ratio was 3:1 (16/6). Initial symptoms were respiratory in about half the patients (10/22). Complete surgical resection was done in five patients. Postoperative cisplatin-based chemotherapy and radiation was administered to seven patients. Pathologic examination showed low grade(n = 14), intermediate grade (n = 7), and high grade (n = 1) TCA. Capsular invasion was present in 83% of the specimens. As of June 2002, nine patients are alive and eight are disease free. The median survival is 44.7 months. Locally invasive disease precluded complete surgical resection in more than half of our cases. Incomplete surgical resection did not preclude long-term survival if multimodality platinum-based therapy was used. PMID- 14758135 TI - Adjuvant cytotoxic and endocrine therapy in pre- and postmenopausal patients with breast cancer and one to nine infiltrated nodes: five-year results of the Hellenic Cooperative Oncology Group randomized HE 10/92 study. AB - SUMMARY: The present randomized phase III trial was designed to detect a 15% benefit in relapse-free survival (RFS) or overall survival (OS) from the incorporation of adjuvant tamoxifen to the combination of CNF [cyclophosphamide, 500 mg/m2; mitoxantrone (Novantrone), 10 mg/m2; fluorouracil, 500 mg/m2 chemotherapy and ovarian ablation in premenopausal patients with node-positive breast cancer and conversely from the incorporation of CNF chemotherapy to adjuvant tamoxifen in node-positive postmenopausal patients. From April 1992 until March 1998, 456 patients with operable breast cancer and one to nine infiltrated axillary nodes entered the study. Premenopausal patients were treated with six cycles of CNF chemotherapy followed by ovarian ablation with monthly injections of triptoreline 3.75 mg for 1 year (Group A, 84 patients) or the same treatment followed by 5 years of tamoxifen (Group B, 92 patients). Postmenopausal patients received 5 years of tamoxifen (Group C, 145 patients) or 6 cycles of CNF followed by 5 years of tamoxifen (Group D, 135 patients). Adjuvant radiation was administered to all patients with partial mastectomy. After a median follow-up period of 5 years, 125 patients (27%) relapsed and 79 (17%) died. The 5-year actuarial RFS for premenopausal patients was 65% in Group A and 68% in Group B (p = 0.86) and for postmenopausal patients 70% in Group C and 67% in Group D (p = 0.36). Also, the respective OS rates were 77% and 80% (p = 0.68) for premenopausal and 84% and 78% (p = 0.10) for postmenopausal patients. Severe toxicities were infrequently seen, with the exception of leukopenia (18%), among the 311 patients treated with CNF. In conclusion, the present study failed to demonstrate a 15% difference in RFS in favor of node-positive premenopausal patients treated with an additional 5 years of tamoxifen after CNF adjuvant chemotherapy and ovarian ablation. Similarly, six cycles of CNF preceding 5 years of tamoxifen did not translate to a 15% RFS benefit in node-positive postmenopausal patients. PMID- 14758137 TI - Second malignancies after chemotherapy and radiotherapy for Hodgkin disease. AB - The purpose of this preliminary study was to determine the incidence of second malignancies after combined-modality therapy for adults with Hodgkin disease and relate it to the details of initial treatment. We retrospectively studied 286 patients ranging in age from 16 to 88 years with stage I or II Hodgkin disease who were treated between 1980 and 1995 with chemotherapy followed 3 to 4 weeks later by radiotherapy. Patients received a median of three cycles of induction chemotherapy. Mitoxantrone, vincristine, vinblastine, and prednisone was used in 161 cases, mechlorethamine, vincristine, procarbazine, and prednisone (MOPP) in 67 cases, Adriamycin, bleomycin, vinblastine, and dacarbazine in 19 cases, lomustine, vinblastine, procarbazine, and prednisone/doxorubicin, bleomycin, dacarbazine, and lomustine in 18 cases, and other chemotherapeutic regimens in the remaining 21 cases. The median radiotherapy dose was 40 Gy given in 20 daily 2-Gy fractions. Median follow-up of surviving patients was 7.4 years. There were 2,230 person-years of observation. Significantly increased relative risks (RR) were observed for acute myeloid leukemia (RR, 69.3; 95% CI, 14.3-202.6) and melanoma (RR, 7.3; 95% CI, 1.5-21.3). The 5-, 10-, and 15-year actuarial risks of acute myeloid leukemia were 0.8%, 1.3%, and 1.3%, respectively. Patients treated with MOPP had the highest 15-year actuarial risk of leukemia (1.6%). The 5-, 10-, and 15-year actuarial risks of solid tumors were 1.9%, 9.3%, and 16.8%, respectively. Consolidative radiotherapy to both sides of the diaphragm resulted in a trend toward an increased risk of solid tumors relative to radiotherapy to only one side of the diaphragm (p = 0.08). In an effort to reduce the risk of second malignancies, we have stopped using the alkylating agents nitrogen mustard and procarbazine and elective paraaortic and splenic radiotherapy after chemotherapy. PMID- 14758138 TI - Potential drug interaction with paclitaxel and highly active antiretroviral therapy in two patients with AIDS-associated Kaposi sarcoma. AB - Highly active antiretroviral therapy (HAART) consisting of multiple combinations of antiretroviral agents that inhibit HIV (HIV) replication has been associated with improvements in CD4+ T-lymphocyte cell counts, suppression of HIV replication, and regression of HIV-associated Kaposi's sarcoma (KS). Several of these agents have complex drug-drug interactions and have the potential to aggravate paclitaxel-associated toxicities. Herein two patients with KS are described who received paclitaxel while on HAART and who developed life threatening toxicities. These cases are presented to alert clinicians to potentially serious drug interactions that can occur between various components of HAART and paclitaxel. Clinicians contemplating taxane-based chemotherapy for HIV-associated KS should carefully monitor patients for adverse events and, depending on their patient's HAART regimen, consider reducing the taxane dose. PMID- 14758139 TI - Pattern of proliferative index (Ki-67) after anti-androgen manipulation reflects the ability of irradiation to control prostate cancer. AB - Anti-androgen (AA) therapy will cause hormone-sensitive prostate cancer cells to undergo apoptosis and/or enter the resting phase of the cell cycle. Although the decrease of tumor burden would be an advantage for tumor control when irradiation is subsequently added, the cells in resting phase would seemingly be less vulnerable to the usual type of radiation-induced cell killing via DNA strand breakage. In this study of patients with prostate cancer, we examined the proliferative index via Ki-67 staining of biopsy material before, during, and after withdrawal of leuprolide. We studied 15 previously untreated patients with locally advanced prostate cancer. Prostate biopsies were taken at three times: 1) initial diagnosis; 2) after 3 consecutive months of intramuscular 7.5 mg depot; and c) 6 weeks after the last dose. External beam radiation (EBRT) then delivered 66 Gy in 33 sessions to local fields. We used the ASTRO definition of prostate specific antigen (PSA) failure. We measured serum luteinizing hormone and total testosterone coinciding with each biopsy date. Immunohistochemical staining was performed using Ki-67 antibody clone MIB-1. The follow-up ranged from 36 to 73 months (median 52 months). We discerned two perturbation patterns of Ki-67 with hormonal manipulation. Pattern 1 demonstrated a drop of Ki-67 labeling after leuprolide was in effect and then after leuprolide withdrawal, the Ki-67 rebounded to less than 120% of baseline. Pattern 2 also showed an initial drop with leuprolide but rebounded to more than 120%. Among eight patients demonstrating pattern 1, only one patient had a PSA failure. In contrast among patients with pattern 2, six of seven failed biochemically (Fisher's exact, p = 0.018). All patients had a LH less than 1.0 during leuprolide effect that rose with its withdrawal. There was no correlation of PSA failure with whether total testosterone did or did not rise to more than 100 ng/dl by the time of the withdrawal phase biopsy. Neither the percent of PSA decline during leuprolide nor the minor PSA rebound 6 to 8 weeks after leuprolide withdrawal correlated with the Ki-67 pattern. The pattern of perturbation of immunohistochemical staining for Ki-67 predicts biochemical failure after moderate-dose EBRT in patients with prostate cancer. Several recent analyses of combined EBRT and AA suggest that some patients may benefit from more prolonged use of AA. Because AA can have substantial side effects and is expensive, a method to select patients likely to benefit from long-term AA would be useful. After neoadjuvant AA manipulation, the Ki-67 perturbation pattern, but not the early PSA changes, may help select patients for long-term AA. The Ki-67 pattern might also be used to select patients needing escalated radiation dosage. Further validation of these concepts beyond this pilot study is suggested. PMID- 14758140 TI - Phase I study with dose escalation of gemcitabine and cisplatin in combination with ifosfamide (GIP) in patients with non-small-cell lung carcinoma. AB - Gemcitabine (G) and cisplatin (P) are active reference agents in patients with non-small-cell lung cancer (NSCLC). Ifosfamide (I) has also been approved for NSCLC treatment. This phase I trial aimed to determine the dose-limiting toxicity (DLT), maximum tolerated dose [maximum tolerated dosage (MTD)], and recommended dose (RD) of a GIP combination in patients with advanced/metastatic NSCLC. In this study, one cycle of chemotherapy combined the following: ifosfamide: 3 g/m2 fixed dose (24-hour intravenous infusion) combined with mesna, day 1; gemcitabine: starting dose 1,000 mg/m2/d, escalating by 250 mg/m2 increments, days 1 and 15; cisplatin: starting dose 80 mg/m2, subsequently 100 mg/m2, day 15; in cohorts of at least 3 patients. Cycles were repeated every 28 days and no hematopoietic growth factors were administered. DLT was evaluated after the first chemotherapy cycle. Thirty-three patients (30 men, 3 women) with stage III (14 patients)/IV (19 patients) NSCLC were treated at eight dose levels, receiving 109 cycles of chemotherapy. Neutropenia was the only DLT reported. Although the MTD was not reached at the highest tested dose level, the RD chosen corresponds to the full doses of the GP3000 doublet standard (G: 3,000 mg/m2; P: 100 mg/m2 per cycle) every 28 days. Nonhematologic toxicities were mainly grade I-II. Relative dose intensities of G, I, and P at the RD were 96%, 98%, and 96%, respectively. Sixteen of 33 patients with measurable/evaluable disease had an objective response including two complete responses. In conclusion, GIP chemotherapy is safe and appears to be active in patients with NSCLC. The RD is gemcitabine: 1,500 mg/m2 days 1 and 15; ifosfamide: 3 g/m2 day 1; cisplatin: 100 mg/m2 day 15. A confirmatory phase II study is currently under way, before a phase III trial of GIP versus GP. PMID- 14758141 TI - Squamous cell carcinoma arising in a second branchial cleft cyst. AB - The existence of primary branchiogenic carcinoma is controversial. In 1950, Martin et al. established four criteria for the diagnosis of primary branchiogenic carcinoma. In 1989, Khafif et al. proposed new criteria, which are currently most recognized in the literature. A pathologic description of the surgical specimen is analyzed and compared with a critical review of the literature. A case is presented in which imaging established the diagnosis of a benign second branchial cleft cyst 2 years prior to the diagnosis of a branchiogenic carcinoma in the same cyst. This cyst was resected and was found to have squamous cell carcinoma arising from the benign epithelium. This case satisfies the histologic criteria established by Martin and Khafif for a primary branchiogenic carcinoma arising in a previously benign second branchial cleft cyst. PMID- 14758142 TI - 5-fluorouracil administered as a 48-hour semiintermittent infusion in combination with leucovorin, cisplatin and epirubicin: phase II study in advanced gastric cancer patients. AB - The main objective of this study was to determine the feasibility and the antitumor activity of a chemotherapy regimen with a 48-hour infusion of 5 fluorouracil (5-FU), leucovorin (LV), cisplatin (CDDP), and epirubicin (EPIDX) administered every 3 weeks in patients with locally advanced or metastatic gastric cancer. Thirty-three patients received CDDP 60 mg/m2 over 30 minutes followed by 1-LV 250 mg/m2 over 2 hours followed by EPIDX 60 mg/m2 over 5 minutes (bolus) and followed by 5-FU 3,800 mg/m2 as a 48-hour semiintermittent continuous infusion, with 67% of total daily dose administered between 4 pm and midnight. Four patients had a locally advanced disease and 29 had metastatic disease. A total of 171 cycles were administered. Most relevant toxicities were stomatitis (grade III in 2% of cycles and 12% of patients) and neutropenia (grade III-IV in 8% of cycles and 28% of patients) with 3 (9%) patients experiencing 1 episode of febrile neutropenia. No toxic deaths occurred. Thirty-one patients were evaluable for response. In 3 patients (9.6%) a complete response and in 11 patients (35.4%) a partial response was observed, for an objective response rate of 45% (95% C.I. 27-64%). Median progression-free and overall survival were 5.9 and 9.8 months, respectively. In conclusion, this regimen is feasible in an outpatient setting with acceptable and manageable toxicities, and it is associated with promising antitumor activity, time to progression, and survival. PMID- 14758143 TI - Cancer management controversy: on residency training. PMID- 14758144 TI - Effect of acute lung injury and acute respiratory distress syndrome on outcome in critically ill trauma patients. AB - OBJECTIVE: Acute lung injury (ALI) and acute respiratory distress syndrome (ARDS) are known to be associated with increased mortality and costs in trauma patients. We estimated the independent impact of these conditions on mortality and cost, beyond the severity of injury with which they are correlated. DESIGN: One-year prospective cohort. PATIENTS AND SETTING: All trauma patients admitted to the intensive care unit in a level I center were evaluated daily for ALI/ARDS using the American-European Consensus Conference definition. MEASUREMENTS AND MAIN RESULTS: The main outcome measures were hospital mortality and costs. Logistic regression was used to model hospital mortality in relation to the presence of ALI and ARDS, adjusting for trauma severity (Injury Severity Score), Acute Physiology Score, and age. Hospital costs were modeled using multivariable linear regression. Of the 1,296 trauma patients surviving beyond the first day, 4% experienced ALI (defined as Pao2/Fio2 of 201-300 mm Hg) and 12% had ARDS (Pao2/Fio2 < or = 200 mm Hg). The crude relative risk of mortality was 2.24 (95% confidence interval, 0.92-5.45) in patients with ALI and 3.84 (95% confidence interval, 2.41-6.13) in patients with ARDS compared with those without ALI/ARDS. However, there was no association of mortality with ALI (relative risk, 0.99; 95% confidence interval, 0.29-3.36) or with ARDS (relative risk, 1.23; 95% confidence interval, 0.63-2.43) after adjustment for age, Injury Severity Score, and Acute Physiology Score. Among patients of comparable age, severity score, and length of stay, median cost was 20% to 30% higher for those with ALI/ARDS. CONCLUSIONS: There is no additional mortality associated with ALI/ARDS above and beyond the factors that can be measured at intensive care unit admission. Therefore, mortality in trauma patients is explained by injury severity at admission and is not affected by the subsequent occurrence of ALI/ARDS. Nonetheless, ALI/ARDS was associated with increased intensive care unit stay and hospital cost, independent of trauma severity. PMID- 14758145 TI - Recombinant human platelet-activating factor acetylhydrolase for treatment of severe sepsis: results of a phase III, multicenter, randomized, double-blind, placebo-controlled, clinical trial. AB - OBJECTIVE: Platelet-activating factor (PAF) and structurally-related oxidized phospholipids are proinflammatory mediators in systemic inflammatory states such as severe sepsis. The enzyme platelet-activating factor acetylhydrolase (PAF-AH) rapidly degrades PAF and oxidized phospholipids into inactive metabolites. Reduced PAF-AH activity has been observed in patients with severe sepsis and may contribute to their systemic inflammatory response and organ dysfunction. A previous clinical trial with recombinant human PAF-AH (rPAF-AH, Pafase) suggested that this treatment may decrease 28-day all-cause mortality in patients with severe sepsis. The current study was undertaken to confirm this result. DESIGN: A prospective, randomized, double-blind, placebo-controlled, multicenter, international trial. SETTING: One hundred forty-six intensive care units from nine countries. PATIENTS: Approximately 2,522 patients were planned to be enrolled < or =12 hrs after the onset of severe sepsis. Eligible patients were randomized to receive either rPAF-AH 1.0 mg/kg or placebo administered intravenously once daily for five consecutive days. MEASUREMENTS AND MAIN RESULTS: The study was terminated based on the recommendation of an independent data and safety monitoring committee after the second of three planned interim analyses, and the enrollment of 1,425 patients. rPAF-AH treatment was well tolerated among the 1,261 patients included in the interim analysis (643 rPAF-AH and 618 placebo), but did not decrease 28-day all-cause mortality compared with placebo (25% for rPAF-AH vs. 24% for placebo; relative risk, 1.03; 95% confidence interval, 0.85-1.25; p =.80). There were no statistically significant differences between treatment groups in any of the secondary efficacy end points. The overall incidence of adverse events was similar among rPAF-AH and placebo-treated patients, and no rPAF-AH-treated patients developed antibodies to PAF-AH. CONCLUSIONS: rPAF-AH was well tolerated and not antigenic, but did not decrease 28-day all-cause mortality in patients with severe sepsis. PMID- 14758146 TI - Treatment with N-acetylcysteine plus deferoxamine protects rats against oxidative stress and improves survival in sepsis. AB - OBJECTIVE: Oxidative stress plays an important role in the development of multiple organ failure and septic shock. Here we have evaluated the effects of a combination of antioxidants (N-acetylcysteine plus deferoxamine) in a murine model of polymicrobial sepsis induced by cecal ligation and puncture (CLP). DESIGN: Prospective, randomized, controlled experiment. SETTING: Animal basic science laboratory. SUBJECTS: Male Wistar rats, weighing 300-350 g. INTERVENTIONS: Rats subjected to CLP were treated with either N-acetylcysteine (20 mg/kg, 3 hrs, 6 hrs, 12 hrs, 18 hrs, and 24 hrs after CLP, subcutaneously) plus deferoxamine (20 mg/kg, 3 hrs and 24 hrs after CLP, subcutaneously) or vehicle with or without "basic support" (saline at 50 mL/kg immediately and 12 hrs after CLP plus ceftriaxone at 30 mg/kg and clindamycin 25 mg/kg every 6 hrs). MEASUREMENTS AND MAIN RESULTS: After 12 hrs, tissue myeloperoxidase (indicator of neutrophil infiltration), thiobarbituric acid reactive species (as a marker of oxidative stress), catalase and superoxide dismutase activities (antioxidant enzymes), and mitochondrial superoxide production (index of uncoupling of electron transfer chain) were measured in major organs involved in septic response. Rats treated with antioxidants had significantly lower myeloperoxidase activity and thiobarbituric acid reactive species formation in all organs studied. Mitochondrial superoxide production was significantly reduced by antioxidant treatment. Furthermore, antioxidants significantly improved the balance between catalase and superoxide dismutase activities. Survival in untreated septic rats was 10%. Survival increased to 40% with fluids and antibiotics. In rats treated only with N-acetylcysteine plus deferoxamine, survival was also significantly improved (47%) in a manner similar to basic support. Survival increased to 66% with basic support with N-acetylcysteine plus deferoxamine. CONCLUSIONS: Our data provide the first experimental demonstration that N-acetylcysteine plus deferoxamine reduces the consequences of septic shock induced by CLP in the rat, by decreasing oxidative stress and limiting neutrophil infiltration and mitochondrial dysfunction, thereby improving survival. PMID- 14758147 TI - Low caloric intake is associated with nosocomial bloodstream infections in patients in the medical intensive care unit. AB - OBJECTIVE: To determine whether caloric intake is associated with risk of nosocomial bloodstream infection in critically ill medical patients. DESIGN: Prospective cohort study. SETTING: Urban, academic medical intensive care unit. PATIENTS: Patients were 138 adult patients who did not take food by mouth for > or =96 hrs after medical intensive care unit admission. MEASUREMENTS: Daily caloric intake was recorded for each patient. Participants subsequently were grouped into one of four categories of caloric intake: <25%, 25-49%, 50-74%, and > or =75% of average daily recommended calories based on the American College of Chest Physicians guidelines. Simplified Acute Physiology Score II and serum albumin were measured on medical intensive care unit admission. Serum glucose (average value and maximum value each day) and route of feeding (enteral, parenteral, or both) were collected daily. Nosocomial bloodstream infections were identified by infection control surveillance methods. MAIN RESULTS: The overall mean (+/-sd) daily caloric intake for all study participants was 49.4 +/- 29.3% of American College of Chest Physicians guidelines. Nosocomial bloodstream infection occurred in 31 (22.4%) participants. Bivariate Cox analysis revealed that receiving > or =25% of recommended calories compared with <25% was associated with significantly lower risk of bloodstream infection (relative hazard, 0.24; 95% confidence interval, 0.10-0.60). Simplified Acute Physiology Score II also was associated with risk of nosocomial bloodstream infection (relative hazard, 1.27; 95% confidence interval, 1.01-1.60). Average daily serum glucose, admission serum albumin, time to initiating nutritional support, and route of nutrition did not affect risk of bloodstream infection. After adjustment for Simplified Acute Physiology Score II in a multivariable analysis, receiving > or =25% of recommended calories was associated with a significantly lower risk of bloodstream infection (relative hazard, 0.27; 95% confidence interval, 0.11 0.68). CONCLUSIONS: In the context of reducing risk of nosocomial bloodstream infections, failing to provide > or =25% of the recommended calories may be harmful. Higher caloric goals may be necessary to achieve other clinically important outcomes. PMID- 14758148 TI - Electronic monitoring and voice prompts improve hand hygiene and decrease nosocomial infections in an intermediate care unit. AB - OBJECTIVE: To determine whether electronic monitoring of hand hygiene and voice prompts can improve hand hygiene and decrease nosocomial infection rates in a surgical intermediate care unit. DESIGN: Three-phase quasi-experimental design. Phase I was electronic monitoring and direct observation; phase II was electronic monitoring and computerized voice prompts for failure to perform hand hygiene on room exit; and phase III was electronic monitoring only. SETTING: Nine-room, 14 bed intermediate care unit in a university, tertiary-care institution. All patient rooms, utility room, and staff lavatory were monitored electronically. PARTICIPANTS: All healthcare personnel including physicians, nurses, nursing support personnel, ancillary staff, all visitors and family members, and any other personnel interacting with patients on the intermediate care unit. All patients with an intermediate care unit length of stay >48 hrs were followed for nosocomial infection. INTERVENTIONS: Electronic monitoring during all phases, computerized voice prompts during phase II only. MEASUREMENTS AND MAIN RESULTS: We evaluated a total of 283,488 electronically monitored entries into a patient room with 251,526 exits for 420 days (10,080 hrs and 3,549 patient days). Compared with phase I, hand hygiene compliance in patient rooms improved 37% during phase II (odds ratio, 1.38; 95% confidence interval, 1.04-1.83) and 41% in phase III (odds ratio, 1.41; 95% confidence interval, 1.07-1.84). When adjusting for patient admissions during each phase, point estimates of nosocomial infections decreased by 22% during phase II and 48% during phase III; when adjusting for patient days, the number of infections decreased by 10% during phase II and 40% during phase III. Although the overall rate of nosocomial infections significantly decreased when combining phases II and III, the association between nosocomial infection and individual phase was not significant. CONCLUSIONS: Electronic monitoring provided effective ongoing feedback about hand hygiene compliance. During both the voice prompt phase and post-intervention phase, hand hygiene compliance and nosocomial infection rates improved suggesting that ongoing monitoring and feedback had both a short-term and, perhaps, a longer-term effect. PMID- 14758149 TI - Does the storage time of transfused red blood cells influence regional or global indexes of tissue oxygenation in anemic critically ill patients? AB - OBJECTIVE: To determine whether transfusion of red cells either < or =5 days or > or =20 days from donation alters tonometric indexes of gastric mucosal oxygenation or global oxygenation parameters in euvolemic anemic critically ill patients without ongoing hemorrhage. The a priori hypothesis was that stored red cells worsen gastric oxygenation. DESIGN: Prospective, double-blind, randomized study. SETTING: A 12-bed general medical/surgical intensive care unit in a Scottish teaching hospital. PATIENTS: Ventilated euvolemic anemic (mean +/- sd hemoglobin, 85.8 +/- 8.4 g/L) critically ill patients with significant organ failure, but no evidence of hemorrhage. INTERVENTIONS: After baseline measurements, patients were randomized to receive two units of leukodepleted red cells that were either < or =5 days (ten patients) or > or =20 days (12 patients) after donation according to a standardized protocol. MEASUREMENTS AND MAIN RESULTS: Changes in gastric to arterial Pco2 gap (Pg-Paco2 gap), gastric intramucosal pH, arterial pH, arterial base excess, and arterial lactate concentrations were measured during baseline (2.5 hrs), during transfusion (3 hrs), and for 5 hrs after transfusion. Mean age of red cells stored < or =5 days was 2 days (first and third quartile, 2, 2.25; range, 2-3); red cells stored >/=20 days had a mean age of 28 days (first and third quartile, 27, 31; range, 22 32). Hemoglobin concentration increased by 15.0 g/L and 16.6 g/L, respectively, in the fresh and stored groups (p =.62). There were no significant differences between the groups either using treatment-by-time analysis or comparing the pre- and posttransfusion periods either for Pg-Paco2 gap (mean difference, 0.03 kPa; 95% confidence limits, -1.66, 1.72) or gastric intramucosal pH (mean difference, 0.015 pH units; 95% confidence limits, -0.054, 0.084). The mean change within each group from the pre- to posttransfusion period for Pg-Paco2 gap and gastric intramucosal pH, respectively, was 0.56 kPa (95% confidence limits, -0.68, 1.79) and -0.018 pH units (95% confidence limits, -0.069, 0.032) for "fresh" red cells and 0.52 kPa (95% confidence limits, -0.6, 1.64) and -0.033 pH units (95% confidence limits, -0.080, 0.129) for "stored" red cells. There was no statistically or clinically significant improvement in any other oxygenation index during the measurement period for either group compared to baseline values. CONCLUSIONS: Transfusion of stored leukodepleted red cells to euvolemic, anemic, critically ill patients has no clinically significant adverse effects on gastric tonometry or global indexes of tissue oxygenation. These findings do not support the use of fresh red cells in critically ill patients. PMID- 14758150 TI - Noninvasive ventilation in patients with acute hypercapnic exacerbation of chronic obstructive pulmonary disease who refused endotracheal intubation. AB - OBJECTIVE: To determine the long-term outcome of noninvasive ventilation in chronic obstructive pulmonary disease patients who refused intubation for acute hypercapnic respiratory failure. DESIGN: Prospective, observational study. SETTING: Noninvasive ventilation unit in an acute regional hospital in Hong Kong. METHODS: The study recruited 37 chronic obstructive pulmonary disease patients who had the do-not-intubate code and developed acute hypercapnic respiratory failure. They were offered noninvasive ventilation, and their long-term outcomes were followed. Survival and event-free survival (an event is death or recurrent acute hypercapnic respiratory failure) were analyzed by survival analysis. Their disease profile and outcome were compared with another 43 chronic obstructive pulmonary disease patients without the do-not-intubate codes, who had acute hypercapnic respiratory failure and received noninvasive ventilation during the study period (usual care group). RESULTS: Patients in the do-not-intubate group were significantly older (p =.029), had worse dyspnea score (p <.001), worse Katz Activities of Daily Living score (p <.001), worse comorbidity score (p =.024), worse Acute Physiology and Chronic Health Evaluation II score (p =.032), lower hemoglobin (p =.001), and longer stay in the hospital during the past year (p =.001) than patients who received usual care. In the do-not-intubate group, the median survival was 179 days, and 1-yr actuarial survival was 29.7%; in the usual care group, the median survival was not reached during follow-up, and 1-yr actuarial survival was 65.1% (p <.0001). In the do-not-intubate group, the median event-free survival was 102 days, and 1-yr event-free survival was 16.2%; in the usual care group, median event-free survival was 292 days, and 1-yr event-free survival was 46.5% (p =.0004). CONCLUSIONS: A 1-yr survival of about 30% was recorded in chronic obstructive pulmonary disease patients with the do-not intubate code who developed acute hypercapnic respiratory failure requiring noninvasive ventilation. The majority of survivors developed another life threatening event in the following year. Information generated from this study is important to physicians and chronic obstructive pulmonary disease patients when they are considering using noninvasive ventilation as a last resort. PMID- 14758151 TI - Life after death: posttraumatic stress disorder in survivors of cardiac arrest- prevalence, associated factors, and the influence of sedation and analgesia. AB - OBJECTIVE: Cardiac arrest is possibly one of the most traumatizing conditions for patients, but to date, its influence on psychic morbidity remains unknown. Posttraumatic stress disorder is a unique symptom configuration after an extreme event consisting of intrusion re-experiencing, avoidance and numbness, and hyperarousal symptoms. We studied a) the prevalence of posttraumatic stress disorder (PTSD) in long term survivors of cardiac arrest; b) the role of specific stress factors related to cardiac arrest for the development of PTSD; and c) the influence of sedation and analgesia during or after cardiac arrest on the occurrence of PTSD. DESIGN: Prospective, cohort study. SETTING: University teaching hospital. PATIENTS: Analysis was performed in cardiac arrest survivors who were discharged with favorable neurologic outcome during an 8-yr period (1991 1999). INTERVENTIONS: All patients received the Davidson Trauma Score for the assessment of PTSD and a modified German version of the EuroQol questionnaire for assessment of quality of life. Cardiac arrest circumstances and administration of sedation and analgesia were assessed. MEASUREMENTS AND MAIN RESULTS: Of 1,630 initially resuscitated patients, 270 patients were discharged with good neurologic outcome. A total of 226 patients were contacted, and 143 patients (63% of all eligible patients) completed the study. Mean time from cardiac arrest to follow up was 45 months (range, 24-66). Thirty-nine patients (27%; 95% confidence interval, 21% to 35%) had a Davidson Trauma Score >40 and fulfilled criteria for PTSD. Patients with PTSD had a significantly lower quality of life. The only independent risk factor for the development of PTSD was younger age. There was no difference between patients with or without PTSD regarding the use of sedation and analgesia during or after cardiac arrest. CONCLUSION: The prevalence of PTSD in cardiac arrest survivors is high. Besides younger age, neither clinical factors nor the use of sedation and analgesia were associated with development of PTSD. PMID- 14758152 TI - Study of clinical course of organ dysfunction in intensive care. AB - OBJECTIVE: Multiple organ dysfunction is a common cause of death in intensive care units. We describe the daily course of multiple organ dysfunction measured by the Sequential Organ Failure Assessment score in a population-based cohort of critically ill patients. DESIGN: Prospective cohort study. SETTING: Adult multisystem intensive care units in the Calgary Health Region. PATIENTS: A total of 1,436 patients admitted from May 1, 2000 to April 30, 2001. MEASUREMENTS: Temporal change in Sequential Organ Failure Assessment score. INTERVENTIONS: None; observational study. MAIN RESULTS: The mean age was 58 yrs (range, 14-100). The mean +/- sd intensive care unit admission Acute Physiology and Chronic Health Evaluation II score was 25 +/- 9. The median intensive care unit length of stay was 4 days (interquartile range, 2-8), and the median hospital length of stay was 15 days (interquartile range, 7-32). A total of 20.5% of patients were infected at admission, and 26.0% were immediately postoperative. Intensive care unit mortality was 27.0%, and hospital mortality was 35.1%. The daily Sequential Organ Failure Assessment score was significantly higher in nonsurvivors than survivors. A population-averaged model determined a mean rate of change of Sequential Organ Failure Assessment score to be -0.29 per day (95% confidence interval, -0.32 to 0.25) for survivors and -0.03 per day (95% confidence interval, -0.08 to 0.03) for nonsurvivors (overall regression, p <.0001). Patients with infection had higher admission Sequential Organ Failure Assessment scores compared with patients without infection (difference, 1.8; p <.001), but a similar rate of daily change. CONCLUSIONS: Multiple organ dysfunction, does not follow a course of progressive and sequential failure. Evidence of differential daily change should further inform the use of organ failure scores as surrogate outcomes in clinical trials. PMID- 14758153 TI - Intensive care unit survivors have fewer hospital readmissions and readmission days than other hospitalized patients in British Columbia. AB - OBJECTIVE: Intensive care unit (ICU) patients who survive their hospital admission have a long-term survival that is similar to that of hospitalized patients who do not require ICU admission. The risk of future readmission to the hospital for these two patient groups is unknown. The objective of this study was to determine the association between ICU admission and number of readmissions to the hospital and number of readmission days. DESIGN: Cohort study for 3 yrs between 1994 and 1997. SETTING: All acute care hospitals in British Columbia, Canada. PATIENTS: A total of 23,859 patients admitted to the ICU and 40,052 patients admitted to the hospital but not the ICU (5% random sample of total). INTERVENTION: None. MEASUREMENTS AND MAIN RESULTS: We measured the number of readmissions to the hospital and the number of readmission days after discharge from the first admission to the hospital during the study period. For survivors to the end of the study period, patients who had been in the ICU had 0.66 readmissions per year and 5.29 readmission days per year compared with 0.73 readmissions per year and 5.48 readmission days per year for control subjects. After controlling for age, sex, socioeconomic status, number of previous ICU and hospital admissions, major clinical category during index admission, comorbidity score during index admission, length of hospital stay during index admission, size of index hospital, and period of follow-up, ICU admission was associated with fewer readmissions (survivors: rate ratio, 0.80; 95% confidence interval, 0.77-0.82; nonsurvivors: rate ratio, 0.85; 95%, confidence interval, 0.82-0.89) and readmission days (survivors: rate ratio, 0.91; 95% confidence interval, 0.87 0.95; nonsurvivors: rate ratio, 0.87; 95%, confidence interval, 0.81-0.92) than admission to the hospital but not the ICU. CONCLUSIONS: Survivors of a hospital stay that includes admission to an ICU have fewer hospital readmissions and readmission days after their discharge than do survivors of a hospital stay without intensive care. PMID- 14758154 TI - Role of circulating cytokines and chemokines in exertional heatstroke. AB - OBJECTIVE: The interplay between inflammatory and anti-inflammatory cytokines, as well as chemokines, has not been well explored in exertional heatstroke. DESIGN: Prospective, observational study. PATIENTS: Seventeen military recruits who developed exertional heatstroke and 17 exertional controls who did not develop exertional heatstroke during the same training exercises. SETTING: University teaching hospital. MEASUREMENTS AND MAIN RESULTS: The severity of exertional heatstroke was evaluated using a Simplified Acute Physiology Score. Plasma cytokines and chemokines were determined using enzyme-linked immunosorbent assay kits. Body temperatures were 41.2 +/- 1.2 degrees C and 37.6 +/- 0.8 degrees C in exertional heatstroke and exertional controls, respectively. Significantly, plasma cytokines including interleukin (IL)-1beta (3.1 +/- 1.6 vs. 1.2 +/- 0.8 pg/mL; p <.05), tumor necrosis factor alpha (4.9 +/- 4.1 vs. 1.2 +/- 2.4 pg/mL; p <.05), IL-6 (15.8 +/- 3.2 vs. 1.2 +/- 1.2 pg/mL; p <.01), interferon gamma (7.3 +/- 4.9 vs. 2.4 +/- 4.1 pg/mL; p <.01), IL-2 receptor (1568 +/- 643 vs. 610 +/- 214 pg/mL; p <.01), IL-4 (2.5 +/- 1.2 vs. 1.2 +/- 0.8 pg/mL; p <.05), and IL-10 (12.9 +/- 9.4 vs. 2.5 +/- 4.9 pg/mL; p <.01) and serum chemokines IL-8 (84.2 +/- 79.9 vs. 10.4 +/- 3.2 pg/mL; p <.01), monocyte chemoattractant protein 1 (959 +/- 589 vs. 158 +/- 217 pg/mL; p <.01), and RANTES (12464 +/- 10505 vs. 5570 +/- 2894 pg/mL; p <.01) were elevated in exertional heatstroke compared with exertional controls. Among cytokines, IL-6, interferon gamma, and IL-2 receptor were positively correlated with Simplified Acute Physiology Score (r =.573, p <.01; r =.625, p <.01; and r =.56, p <.05, respectively). Among chemokines, only serum monocyte chemoattractant protein 1 was positively correlated with Simplified Acute Physiology Score (r =.78, p <.001). There was no correlation between either cytokines or chemokines and body temperature. CONCLUSIONS: Proinflammatory cytokines IL-1beta, tumor necrosis factor alpha, IL-6; T helper 1 cytokines INF gamma and IL-2 receptor; and chemokines IL-8, monocyte chemoattractant protein 1, and RANTES are increased in patients with exertional heatstroke. T helper 2 cytokines may play a role as anti-inflammatory cytokines. IL-6, interferon gamma, IL-2 receptor, and monocyte chemoattractant protein 1 may serve as prognostic indicators of disease severity in exertional heatstroke. PMID- 14758155 TI - Hypothalamic-pituitary-adrenal axis dysfunction in critically ill patients with traumatic brain injury: incidence, pathophysiology, and relationship to vasopressor dependence and peripheral interleukin-6 levels. AB - OBJECTIVE: To investigate hypothalamic-pituitary-adrenal axis function in patients requiring mechanical ventilation for traumatic brain injury and to assess the relation of hypothalamic-pituitary-adrenal axis abnormalities with vasopressor dependence and peripheral cytokine levels. DESIGN: Prospective study. SETTING: General intensive care unit in a university teaching hospital. PATIENTS: Forty patients (33 men and 7 women) with moderate to severe traumatic brain injury (mean age, 37 +/- 16 yrs) were studied the day after termination of mechanical ventilation (7-60 days after trauma). INTERVENTIONS: First, a morning blood sample was obtained to measure baseline cortisol, corticotropin, interleukin-6, and tumor necrosis factor alpha. Subsequently, 1 microg of synthetic corticotropin was injected intravenously, and 30 mins later, a second blood sample was drawn to determine stimulated plasma cortisol. Based on data derived from healthy volunteers, patients having stimulated cortisol levels <18 microg/dL were defined as nonresponders to the low-dose stimulation test. Thirty one patients underwent also a human corticotropin releasing hormone test. MEASUREMENTS AND MAIN RESULTS: In traumatic brain injury patients, mean baseline and low-dose stimulation test-stimulated cortisol levels were 17.2 +/- 5.4 microg/dL and 24.0 +/- 6.6 microg/dL, respectively. The median increment in cortisol was 5.9 microg/dL. Basal corticotropin levels ranged from 3.9 to 118.5 pg/mL. Six of the 40 patients (15%) failed the low-dose stimulation test. The human corticotropin releasing hormone test (performed in 26 responders and five nonresponders) revealed diminished cortisol release only in the low-dose stimulation test nonresponder patients. Corticotropin responses to corticotropin releasing hormone were consistent with both primary (three patients) and/or secondary (two patients) adrenal dysfunction. In retrospect, nonresponders to the low-dose stimulation test more frequently required vasopressors (6/6 [100%] vs. 16/34 [47%]; p =.02) and for a longer time interval (median, 0 vs. 293 hrs; p =.006) compared with responders. Furthermore, nonresponders had higher interleukin-6 levels compared with responders (56.03 vs. 28.04 pg/mL; p =.01), whereas tumor necrosis factor alpha concentrations were similar in the two groups (2.42 vs. 1.55 pg/mL; p =.53). CONCLUSIONS: Adrenal cortisol secretion after dynamic stimulation is deficient in a subset of critically ill patients with moderate to severe head injury. This disorder is associated with prior vasopressor dependency and higher interleukin-6 levels. PMID- 14758156 TI - Proportional-assist ventilation compared with pressure-support ventilation during exercise in volunteers with external thoracic restriction. AB - OBJECTIVES: Proportional-assist ventilation (PAV) is able to unload respiratory muscles in proportion to the subject's inspiratory effort. However, leak-related alterations in the flow signal, effort-induced modifications in respiratory mechanics, or approximate adjustment of PAV could jeopardize such a theory. The aim of this study was to compare noninvasive PAV and pressure-support ventilation (PSV) in healthy volunteers with external thoracic restriction at rest and during exercise. DESIGN: Prospective, crossover, randomized study. SETTING: Investigation unit in a nonteaching hospital. PATIENTS: Seven volunteers with external thoracic restriction. INTERVENTION: After external thoracic restriction to increase elastance (9.00 +/- 1.63 cm H2O/L estimated from the level of elastic assistance), PAV and PSV were compared at rest and during exercise (90 W for 10 mins). MEASUREMENTS AND MAIN RESULTS: Flow, airway pressure, and changes in esophageal pressure were measured, and the tidal volume (Vt) and inspiratory muscle effort indexes were calculated. At rest, all variables were comparable during PSV and PAV. Exercise produced a 200% increased in Vt with no change in the breathing frequency and a 400% increased in inspiratory muscle effort indexes. During exercise, peak inspiratory airway pressure was significantly higher with PAV than with PSV (24 +/- 5 vs. 10 +/- 2 cm H2O, p <.05). The Vt and breathing frequency (23 +/- 4 vs. 24 +/- 3 breaths/min) were similar, but the inspiratory muscle effort indexes were significantly lower with PAV than with PSV. A significant linear correlation was found between changes in esophageal pressure and the peak inspiratory airway pressure during PAV (r =.94, p =.0001), whereas, as expected, it was not the case during PSV (r =.27, p =.34). CONCLUSION: In volunteers with external thoracic restriction mimicking a patient with increased elastic work of breathing, the breathing pattern at rest and during exercise were comparable with PSV and PAV, whereas inspiratory muscle effort was lower with PAV during exercise because of the significant automatic increase in assistance with PAV. PMID- 14758157 TI - Limitation of life support: frequency and practice in a Hong Kong intensive care unit. AB - OBJECTIVE: To examine the frequency and the decision-making processes involved in limiting (withdrawing and withholding) life support therapy in critically ill Chinese patients in the intensive care unit. DESIGN: Prospective survey of patients who had life support limited between April 1997 and March 1999. SETTING: Medical and surgical intensive care unit of a teaching hospital. PATIENTS: All patients admitted to the intensive care unit of the Prince of Wales Hospital who subsequently died and/or had life support limited. Brain-dead patients were excluded from analysis. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Of 490 patients who died in the intensive care unit, limitation of life support occurred in 288 (58.8%). Relatives or patients requested limitation of life support in 32 cases (11%). The family and/or patient concurred with limitation of life support in 273 occasions (95%). Therapy was withheld in 30.8% and withdrawn in 28.0% of deaths. Therapy limited included inotropes, additional oxygen, and renal replacement therapy. CONCLUSIONS: Limitation of therapy in dying Chinese patients occurs frequently in intensive care patients, and both patients and relatives concur with medical decisions to limit therapy in these patients. Withholding therapy rather than withdrawing therapy occurs more frequently than in Western populations. PMID- 14758158 TI - Pain behaviors observed during six common procedures: results from Thunder Project II. AB - OBJECTIVE: Patients frequently display behaviors during procedures that may be pain related. Clinicians often rely on the patient's demonstration of behaviors as a cue to presence of pain. The purpose of this study was to identify specific pain-related behaviors and factors that predict the degree of behavioral responses during the following procedures: turning, central venous catheter insertion, wound drain removal, wound care, tracheal suctioning, and femoral sheath removal. DESIGN: Prospective, descriptive study. SETTING: Multiple units in 169 hospitals in United States, Canada, England, and Australia. PATIENTS: A total of 5,957 adult patients who underwent one of the six procedures. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: A 30-item behavior observation tool was used to note patients' behaviors before and during a procedure. By comparing behaviors exhibited before and during the procedure as well as behaviors in those with and without procedural pain (as noted on a 0-10 numeric rating scale), we identified specific procedural pain behaviors: grimacing, rigidity, wincing, shutting of eyes, verbalization, moaning, and clenching of fists. On average, there were significantly more behaviors exhibited by patients with vs. without procedural pain (3.5 vs. 1.8 behaviors; t = 38.3, df = 5072.5; 95% confidence interval, 1.6-1.8). Patients with procedural pain were at least three times more likely to have increased behavioral responses than patients without procedural pain. A simultaneous regression model determined that 33% of the variance in amount of pain behaviors exhibited during a procedure was explained by three factors: degree of procedural pain intensity, degree of procedural distress, and undergoing the turning procedure. CONCLUSIONS: Because of the strong relationship between procedural pain and behavioral responses, clinicians can use behavioral responses of verbal and nonverbal patients to plan for, implement, and evaluate analgesic interventions. PMID- 14758159 TI - Medication errors involving continuously infused medications in a surgical intensive care unit. AB - OBJECTIVE: To document the incidence of medication errors related to medications administered by continuous infusion. DESIGN: Observational study. SETTING: Sixteen-bed surgical intensive care unit. MEASUREMENTS AND MAIN RESULTS: All continuous infusions in the surgical intensive care unit were evaluated at least once daily for correct flow-sheet charting, concentration, infusion rate, and dose administered, as well as patients' heights and weights (actual, ideal, and "dry"). Collected information was examined to determine the error rate, types of errors occurring, and weight used for dose calculation. Variations inpatient weight measures were compared. Seventy-one patients with 202 total infusions were observed. Errors involving continuously infused medications in our surgical intensive care unit occurred at a rate of 105.9 per 1,000 patient days. For nonweight-based infusions, 94% of doses were delivered correctly. Slightly >10% of the doses administered for weight-based infusions (dose based on dry body weight) were incorrect. Significant differences were found between the weight measurements recorded, but this did not translate into statistically significant differences in the apparent calculated doses delivered. CONCLUSIONS: Medications delivered by continuous infusion, particularly those that are weight based, can contribute to medication errors in the intensive care unit. A large proportion (87.6%) of doses for weight-based infusions was calculated based on estimated or unreliable admission weights. There were no severe consequences resulting from the errors observed in this 1 month investigation; however, depending on the pharmacokinetic characteristics of the drug being administered, there is a potential to deliver artificially low or high doses resulting in subtherapeutic or adverse effects. PMID- 14758160 TI - Plasma interferon-gamma and interleukin-10 concentrations in systemic meningococcal disease compared with severe systemic Gram-positive septic shock. AB - OBJECTIVE: To analyze plasma interferon-gamma and interleukin-10 concentrations in patients with systemic meningococcal disease and patients with severe Gram positive septic shock caused by Streptococcus pneumoniae or Staphylococcus aureus. To study the in vitro cytokine (interferon-gamma and interleukin-10) responses in a whole blood model boosted with heat-killed Neisseria meningitidis, S. pneumoniae, and S. aureus before and after treatment with recombinant interleukin-10 or recombinant interferon-gamma. DESIGN: Experimental study. SETTING: Laboratory. SUBJECTS: Plasma samples were collected from patients with systemic meningococcal disease (n = 66) and patients with severe Gram-positive septic shock caused by S. pneumoniae (n = 4) or S. aureus (n = 3). INTERVENTIONS: Whole blood was boosted with heat-killed N. meningitidis, S. pneumoniae, and S. aureus (1 x 106 colony forming units/mL), and plasmas were analyzed for interleukin-10 or interferon-gamma at 0, 5, 12, and 24 hrs. Furthermore, recombinant interleukin-10 or recombinant interferon-gamma was added before bacteria, and the effect on the secretion of interferon-gamma and interleukin-10, respectively, was analyzed after 24 hrs. MEASUREMENTS AND MAIN RESULTS: The median concentration of interferon-gamma was 15 pg/mL and of interleukin-10 was 10,269 pg/mL in patients with meningococcal septic shock (n = 24) compared with median interferon-gamma concentration of 3400 pg/mL and interleukin-10 concentration of 465 pg/mL in patients with severe Gram-positive shock (p =.001). Increased interferon-gamma concentrations were associated with case fatality (p =.011). In a whole blood model we demonstrated that 1 x 106 colony forming units/mL of N. meningitidis induced more interleukin-10 but less interferon-gamma than S. pneumoniae. S. aureus induced minimal secretion of both cytokines. Recombinant interleukin-10 efficiently down-regulated the secretion of interferon gamma, and vice versa, as shown in a whole blood model. CONCLUSION: We speculate whether high concentrations of interleukin-10 contribute to the low concentrations of interferon-gamma in fulminant meningococcal septicemia. In addition, it appears as if interferon-gamma plays a minor role in the pathophysiology of meningococcal septic shock. PMID- 14758161 TI - Transfusion-related leukocytosis in critically ill patients. AB - OBJECTIVE: We observed that many critically ill patients developed leukocytosis following blood transfusions. To validate this observation and to explore a possible mechanism, a prospective study was designed. DESIGN: Prospective, non interventional study. SETTING: Surgical/medical intensive care unit in a university-affiliated community hospital. PATIENTS: Consecutive patients who required packed red blood cells transfusion. INTERVENTIONS: White blood cell count (mean +/- SD) x 10(9)/L before and 2, 4, 6, 12, and 24 hrs following transfusion of non-filtered packed red cells was measured in 96 patients. MEASUREMENTS AND MAIN RESULTS: Twenty patients were septic at the time of transfusion, whereas 76 were not. The incidence of post-transfusion leukocytosis in septic vs. nonseptic patients was 15% vs. 76%, respectively (p <.001). The white blood cell count in nonseptic patients increased from 14.3 +/- 4.8 before transfusion to 19.5 +/- 7.0 2 hrs following transfusion (p <.001) and returned to baseline in 24 hrs. In the septic group, no significant post-transfusion leukocytosis occurred. In 11 nonseptic patients requiring more than one unit of packed red cells, a significant increase in mean white blood cell count occurred 2 hrs after transfusion with non-filtered packed red cells, whereas transfusion with pre-storage-filtered packed red cells did not result in such an increase. Interleukin-8 concentrations (pg/mL) in stored non-filtered packed red cells were significantly higher after 4 wks of storage (745.5 +/- 710, p =.02) than at weeks 1 (61.2 +/- 21.6) and 2 (59.3 +/- 29). In the last 16 nonseptic patients, the units of non-filtered packed red cells were assayed for interleukin-8 immediately before transfusion. Interleukin-8 concentrations were higher in units that caused leukocytosis in the recipients compared with those that did not (408.4 +/- 202 vs. 65.1 +/- 49, p =.02). CONCLUSIONS: Transfusion of non-filtered packed red cells, but not of pre-storage-filtered packed red cells, may frequently cause an acute and transient leukocytosis in critically ill nonseptic patients. Interleukin-8 accumulating in stored non-filtered packed red cells may play a role in this phenomenon. Recognition of post packed red cell transfusion leukocytosis may avoid unnecessary investigations and therapies in false suspicion of sepsis. PMID- 14758162 TI - Open lung ventilation improves gas exchange and attenuates secondary lung injury in a piglet model of meconium aspiration. AB - OBJECTIVE: Previous studies failed to show clear benefits of high-frequency ventilation compared with conventional positive pressure ventilation (PPV(CON)) in experimental meconium aspiration syndrome. However, none of these studies applied an open lung ventilation strategy (OLC), which aims to reduce intrapulmonary shunt due to alveolar collapse. We hypothesized that, if combined with an open lung strategy, both high-frequency oscillatory ventilation (HFOV(OLC)) and positive pressure ventilation (PPV(OLC)) would improve gas exchange and attenuate ventilator-induced lung injury in experimental meconium aspiration syndrome. DESIGN: Prospective, randomized animal study. SETTING: Research laboratory of a large university. SUBJECTS: Forty-two newborn piglets. INTERVENTIONS: Thirty minutes after intratracheal meconium instillation, 36 newborn piglets were assigned to one of three ventilation groups-PPV(OLC), HFOV(OLC), or PPV(CON)-and ventilated for 5 hrs. In both OLC groups, collapsed alveoli were actively recruited and thereafter stabilized using the lowest possible airway pressures. During PPV(CON), ventilator settings were adjusted to prevent critical hypoxia (Pao2 <60 torr [8 kPa]). Six animals served as saline controls. MEASUREMENTS AND MAIN RESULTS: Compared with the PPV(CON) group, arterial oxygenation and lung mechanics were superior in both OLC groups and the saline controls. Analysis of the bronchoalveolar lavage fluid obtained after 5 hrs of ventilation showed increased myeloperoxidase activity in the PPV(CON) group compared with both OLC groups and saline controls. Alveolar protein influx was not different between the groups. Histologic analysis revealed a higher lung injury score in the PPV(CON) group compared with the PPV(OLC) and the HFOV(OLC) groups. CONCLUSIONS: Application of the OLC during PPV and HFOV is feasible in experimental meconium aspiration syndrome and results in superior oxygenation and less ventilator-induced lung injury compared with PPV(CON). PMID- 14758163 TI - Use of artificial intelligence to identify cardiovascular compromise in a model of hemorrhagic shock. AB - OBJECTIVE: To determine whether a prototype artificial intelligence system can identify volume of hemorrhage in a porcine model of controlled hemorrhagic shock. DESIGN: Prospective in vivo animal model of hemorrhagic shock. SETTING: Research foundation animal surgical suite; computer laboratories of collaborating industry partner. SUBJECTS: Nineteen, juvenile, 25- to 35-kg, male and female swine. INTERVENTIONS: Anesthetized animals were instrumented for arterial and systemic venous pressure monitoring and blood sampling, and a splenectomy was performed. Following a 1-hr stabilization period, animals were hemorrhaged in aliquots to 10, 20, 30, 35, 40, 45, and 50% of total blood volume with a 10-min recovery between each aliquot. Data were downloaded directly from a commercial monitoring system into a proprietary PC-based software package for analysis. MEASUREMENTS AND MAIN RESULTS: Arterial and venous blood gas values, glucose, and cardiac output were collected at specified intervals. Electrocardiogram, electroencephalogram, mixed venous oxygen saturation, temperature (core and blood), mean arterial pressure, pulmonary artery pressure, central venous pressure, pulse oximetry, and end-tidal CO(2) were continuously monitored and downloaded. Seventeen of 19 animals (89%) died as a direct result of hemorrhage. Stored data streams were analyzed by the prototype artificial intelligence system. For this project, the artificial intelligence system identified and compared three electrocardiographic features (R-R interval, QRS amplitude, and R S interval) from each of nine unknown samples of the QRS complex. We found that the artificial intelligence system, trained on only three electrocardiographic features, identified hemorrhage volume with an average accuracy of 91% (95% confidence interval, 84-96%). CONCLUSIONS: These experiments demonstrate that an artificial intelligence system, based solely on the analysis of QRS amplitude, R R interval, and R-S interval of an electrocardiogram, is able to accurately identify hemorrhage volume in a porcine model of lethal hemorrhagic shock. We suggest that this technology may represent a noninvasive means of assessing the physiologic state during and immediately following hemorrhage. Point of care application of this technology may improve outcomes with earlier diagnosis and better titration of therapy of shock. PMID- 14758164 TI - Rosiglitazone, a ligand of the peroxisome proliferator-activated receptor-gamma, reduces the development of nonseptic shock induced by zymosan in mice. AB - OBJECTIVE: Peroxisome proliferator-activated receptors (PPARs) are members of the nuclear hormone receptor superfamily of ligand-activated transcription factors that are related to retinoid, steroid, and thyroid hormone receptors. The PPAR gamma receptor subtype appears to play a pivotal role in the regulation of cellular proliferation and inflammation. Rosiglitazone (Avandia) is a PPAR-gamma agonist (the most potent PPAR-gamma agonist of the thiazolidinedione antidiabetics). In the present study, we investigated the effects of rosiglitazone on the development of nonseptic shock caused by zymosan in mice. DESIGN: Experimental study. SETTING: University laboratory. SUBJECTS: Male CD mice. INTERVENTIONS: We investigated the effects of rosiglitazone (3 mg/kg) on the development of nonseptic shock caused by zymosan (500 mg/kg, administered intraperitoneally as a suspension in saline) in mice. MEASUREMENTS AND MAIN RESULTS: Organ failure and systemic inflammation in rats were assessed 18 hrs after administration of zymosan and/or rosiglitazone and monitored for 12 days (for loss of body weight and mortality rate). Treatment of mice with rosiglitazone (3 mg/kg intraperitoneally, 1 and 6 hrs after zymosan) attenuated the peritoneal exudation and the migration of polymorphonuclear cells caused by zymosan. Rosiglitazone also attenuated the lung, liver, and pancreatic injury and renal dysfunction caused by zymosan as well as the increase in myeloperoxidase activity and malondialdehyde concentrations caused by zymosan in the lung and intestine. Immunohistochemical analysis for inducible nitric oxide synthase, nitrotyrosine, and poly(adenosine diphosphate-ribose) revealed positive staining in lung and intestine tissues obtained from zymosan-treated mice. The degree of staining for nitrotyrosine, inducible nitric oxide synthase, and poly(adenosine diphosphate-ribose) was markedly reduced in tissue sections obtained from zymosan treated mice that received rosiglitazone. To elucidate whether the protective effects of rosiglitazone are related to activation of the PPAR-gamma receptor, we also investigated the effect of a PPAR-gamma antagonist, GW 9662, on the protective effects of rosiglitazone. GW 9662 (1 mg/kg administered intraperitoneally 30 mins before treatment with rosiglitazone) significantly antagonized the effect of the PPAR-gamma agonist and thus abolished the protective effect. CONCLUSIONS: This study provides evidence, for the first time, that rosiglitazone attenuates the degree of zymosan-induced nonseptic shock in mice. PMID- 14758165 TI - Assessment of the role of antibiotics and enterococcal virulence factors in a mouse model of extraintestinal translocation. AB - OBJECTIVE: To study the relative contribution of antibiotics and bacterial virulence factors in the process of translocation of Enterococcus faecalis from the gut to extraintestinal organs. DESIGN: Prospective controlled animal study. SETTING: Animal experimental laboratory at a university medical center. SUBJECTS: Fifty-two female Balb/c mice. INTERVENTIONS: We developed a mouse model to study the translocation of Enterococcus faecalis from the intestinal tract. Balb/c mice received sterile drinking water or antibiotic combinations to deplete their indigenous intestinal microflora. The animals subsequently were fed genetically engineered enterococci expressing different combinations of the putative enterococcal virulence factors aggregation substance and binding substance. Animals were killed, and their livers, spleens, and mesenteric lymph nodes were aseptically removed and cultured along with fecal samples for enumeration of bacteria. MEASUREMENTS AND MAIN RESULTS: All animals were colonized with the test strains at 2-6 x 109 colony forming units/g of feces; in the antibiotic-treated animals, feces were free from anaerobes and Enterobacteriaceae. In animals fed the identical bacterial mutant, the colony counts in mesenteric lymph nodes were significantly lower in mice not treated with antibiotics than in those treated with antibiotics (p =.016). Multigroup analysis of variance revealed no significant differences of the translocation frequencies for the different mutant strains; however, the differences were statistically significant for all groups receiving antibiotics vs. the group not receiving antibiotics (p <.05-.01). There was a trend (although not statistically significant) for a higher proportion of positive cultures from either spleen or liver in mice that had enterococci recovered from their mesenteric lymph nodes (28%) relative to those that did not have enterococci isolated from the lymph nodes (12%; rate ratio 2.39, p =.30 by logistic regression analysis). CONCLUSIONS: Oral antibiotics can select for extraintestinal translocation of Enterococcus faecalis, and neither aggregation substance nor binding substance seems to be required for this process. The experiments encourage further exploration of host and microbial factors contributing to translocation and may provide a better understanding of the pathogenesis of enterococcal infections in patients in intensive care units. PMID- 14758166 TI - 5-Aminoisoquinolinone, a novel inhibitor of poly(adenosine disphosphate-ribose) polymerase, reduces microvascular liver injury but not mortality rate after hepatic ischemia-reperfusion. AB - OBJECTIVE: The aim of this study was to investigate the impact of the novel, potent, water-soluble inhibitor of poly(adenosine diphosphate-ribose) polymerase (PARP) 5-aminoisoquinolinone (5-AIQ) on hepatic microcirculation, hepatocellular injury, and survival in a murine model of hepatic ischemia-reperfusion. DESIGN: Randomized animal study. SETTING: Research laboratory. SUBJECTS: C57BL6 mice were subjected to warm either partial (90 mins) or total (75 mins) ischemia of the liver. INTERVENTIONS: Either PARP inhibitor 5-AIQ (3 mg/kg) or vehicle was administered to mice intravenously immediately before the start of reperfusion. Sham-operated animals served as controls. MEASUREMENTS AND MAIN RESULTS: As shown by intravital fluorescence microscopy after 30-60 mins of reperfusion, ischemia reperfusion significantly enhanced platelet- and leukocyte-endothelial cell interactions in hepatic microvessels and impaired sinusoidal perfusion. Hepatocellular injury was characterized by an increase in the number of necrotic and apoptotic cells, dramatic elevation of aspartate aminotransferase/alanine aminotransferase serum activity, and lipid peroxidation in liver tissue. 5-AIQ treatment attenuated ischemia-reperfusion-induced increases in the numbers of adherent platelets and leukocytes as well as of necrotic and apoptotic cells and ameliorated perfusion failure. Furthermore, PARP inhibition prevented the increase in aspartate aminotransferase activity after ischemia-reperfusion but did not affect postischemic alanine aminotransferase release. However, no protective impact of 5-AIQ on postischemic oxidative stress was observed. Although PARP inhibition did not alter the survival percentage after ischemia reperfusion (22% in both groups), this approach prolonged survival from 1 to 24 hrs (ischemia-reperfusion + vehicle) up to 48-72 hrs in the treated group. CONCLUSIONS: PARP inhibition with 5-AIQ during hepatic ischemia-reperfusion attenuates microvascular injury and reduces the extent of necrotic/apoptotic cell damage but does not protect from oxidative injury and does not improve postoperative survival rate. PMID- 14758167 TI - Abnormal permeability of inner and outer mitochondrial membranes contributes independently to mitochondrial dysfunction in the liver during acute endotoxemia. AB - OBJECTIVE: This study was designed to determine the role played by the mitochondrial permeability transition in the pathogenesis of mitochondrial damage and dysfunction in a representative systemic organ during the acute phase of endotoxemia. DESIGN: A well-established, normotensive feline model was employed to determine whether pretreatment with cyclosporine A, a potent inhibitor of the mitochondrial permeability transition, normalizes mitochondrial ultrastructural injury and dysfunction in the liver during acute endotoxemia. SETTING: The Ohio State University Medical Center research laboratory. SUBJECTS: Random source, adult, male conditioned cats. INTERVENTIONS: Hemodynamic resuscitation and maintenance of acid-base balance and tissue oxygen availability were provided, as needed, to minimize the potentially confounding effects of tissue hypoxia and/or acidosis on the experimental results. Treatment groups received isotonic saline vehicle (control; n = 6), lipopolysaccharide (3.0 mg/kg, intravenously; n = 8), or cyclosporine A (6.0 mg/kg, intravenously; n = 6) or tacrolimus (FK506, 0.1 mg/kg, intravenously; n = 4) followed in 30 mins by lipopolysaccharide (3.0 mg/kg, intravenously). Liver samples were obtained 4 hrs posttreatment, and mitochondrial ultrastructure, function, and cytochrome c, Bax, and ceramide contents were assessed. MEASUREMENTS AND MAIN RESULTS: As expected, significant mitochondrial injury was apparent in the liver 4 hrs after lipopolysaccharide treatment, despite maintenance of regional tissue oxygen availability. Namely, mitochondria demonstrated high-amplitude swelling and exhibited altered respiratory function. Cyclosporine A pretreatment attenuated lipopolysaccharide induced mitochondrial ultrastructural abnormalities and normalized mitochondrial respiratory control, reflecting protection against inner mitochondrial membrane damage. However, an abnormal permeability of outer mitochondrial membranes to cytochrome c was observed in all lipopolysaccharide-treated groups and was associated with increased mitochondrial concentrations of Bax and ceramide. CONCLUSIONS: These studies confirm that liver mitochondria are early targets of injury during endotoxemia and that inner and outer mitochondrial membrane damage occurs through different mechanisms. Inner mitochondrial membrane damage appears to relate to the mitochondrial permeability transition, whereas outer mitochondrial membrane damage can occur independent of the mitochondrial permeability transition. Preliminary evidence suggests that Bax may participate in lipopolysaccharide-induced outer mitochondrial membrane damage, but further investigations are needed to confirm this. PMID- 14758168 TI - Antibiotics delay but do not prevent bacteremia and lung injury in murine sepsis. AB - OBJECTIVES: To investigate the effect of antibiotics on infection, lung injury, and mortality rate in polymicrobial sepsis and to determine whether an association exists between infection and lung injury and mortality rate. To circumvent the effect of antibiotics on cultures, we used polymerase chain reaction to detect bacteria. DESIGN: Prospective, randomized, controlled laboratory trial. SETTING: University research laboratory. SUBJECTS: C57/BL6 mice. INTERVENTIONS: Mice underwent cecal ligation and puncture without antibiotics (CLP) or with imipenem (CLP + Abx). MEASUREMENTS AND MAIN RESULTS: CLP resulted in 50% mortality rate at 48 hrs and 100% mortality rate at 84 hrs. Antibiotics delayed these time points to 72 and 120 hrs, respectively. Lung injury occurred before mortality in both groups. Polymerase chain reaction detected bacteria in the blood and lungs of all CLP mice by 24 hrs. Antibiotics delayed but did not prevent infection in CLP + Abx mice. Serum tumor necrosis factor-alpha and lung endotoxin were elevated to similar concentrations in both CLP and CLP + Abx mice. CONCLUSIONS: In this model of sepsis, antibiotics delay but do not prevent acute lung injury and mortality. Even in the presence of antibiotics, acute lung injury is strongly associated with bacteremia and bacteria within the lungs. PMID- 14758169 TI - Endotoxin-induced myocardial dysfunction: evidence for a role of sphingosine production. AB - OBJECTIVE: To determine whether sphingomyelinase pathway activation would participate in myocardial depression induced by endotoxin. DESIGN: Randomized, controlled trial. SETTING: Experimental laboratory. SUBJECTS: Male Sprague-Dawley rats, isolated rat heart, and cardiac myocytes. INTERVENTIONS: Cardiovascular function was evaluated in rats injected with saline, endotoxin (10 mg/kg, intravenously), and N-oleoylethanolamine (NOE; 10 mg/kg, intravenously). In ex vivo experiments, isolated rat hearts were perfused with endotoxin (5 microg/mL). For pharmacologic intervention, NOE (1 micromol/L) was admixed to the perfusate 20 mins before endotoxin. In in vitro experiments, ventricular myocytes were incubated with sphingosine (20 microM). Myocyte cell shortening and calcium transient were measured. Mitochondrial membrane potential was measured using the cationic dye tetramethylrhodamine methylester fluorescence technique. MEASUREMENTS AND MAIN RESULTS: Endotoxin treatment at 4 hrs did not alter mean arterial pressure and abdominal blood flow compared with control rats. Left ventricle developed pressure (LVDP) and its first derivatives (i.e., maximal and minimal change in pressure over time [dP/dtmax and dP/dtmin]) were decreased after 4 hrs in endotoxin-treated rats compared with control rats. NOE (10 mg/kg) treatment largely prevented left ventricular systolic function alterations of endotoxin-treated hearts (n = 6 in each group). In isolated rat heart, endotoxin (5 microg/mL) caused increases in tumor necrosis factor-alpha perfusate concentration and delayed depression of LVDP, dP/dtmax, and dP/dtmin after 60 mins, which was partially abrogated in the presence of the ceramidase inhibitor NOE (1 micromol/L). Sphingosine (20 microM) caused decreases in cell fractional shortening, calcium transient, and mitochondrial membrane potential of cardiac myocytes. CONCLUSION: These observations suggest that the sphingomyelinase pathway participates in endotoxin-induced myocardial depression. PMID- 14758170 TI - Down-regulation of hepatic CYP1A2 plays an important role in inflammatory responses in sepsis. AB - OBJECTIVE: Although hepatic cytochrome P-450 protein concentrations are altered following endotoxin shock, changes in P-450 isoforms in sepsis have not been fully investigated. The aim of this study was to determine whether the major P 450 isoform in rat liver (i.e., CYP1A2) is down-regulated during the progression of sepsis and, if so, whether reduction of P-450 enzyme system plays an important role in the inflammatory response. DESIGN: Prospective, controlled, and randomized animal study. SETTING: A university/institute research laboratory. SUBJECTS: Male adult Sprague-Dawley rats were subjected either to polymicrobial sepsis by cecal ligation and puncture (CLP) or to sham operation followed by the administration of normal saline solution (i.e., fluid resuscitation). INTERVENTIONS: P-450 isoforms in the liver (i.e., CYP1A2 and 4A1) were determined using reverse transcription polymerase chain reaction and Western blot analysis at various time points after CLP. MEASUREMENTS AND MAIN RESULTS: The results indicate that CYP1A2 messenger RNA expression decreased significantly at 10 and 20 hrs whereas its protein concentrations decreased at 20 hrs after the induction of sepsis. In contrast, CYP4A1 messenger RNA and protein concentrations were not altered even at 20 hrs after CLP. In an additional experiment, all P-450 isoforms were inhibited by pretreatment with 1-aminobenzotriazole to determine the effect of cytochrome P-450 blockade on inflammatory responses by assessing proinflammatory cytokines. The results show further increases in serum concentrations of tumor necrosis factor-alpha, interleukin-1beta, and interleukin 6 in aminobenzotriazole-treated animals at 10 hrs after CLP, which was associated with elevated concentrations of circulating lactate and severe morphologic alterations in the liver. These results suggest that the integrity of the cytochrome P-450 enzyme system plays an important role in septic inflammatory response. CONCLUSION: The major hepatic P-450 isoform CYP1A2 is down-regulated and inhibition of P-450 enzyme system is associated with an exacerbated inflammatory response in sepsis. Treatment with pharmaceutical agents that regulate or are metabolized by P-450 enzymes might be approached cautiously in the septic patient if this holds true in a clinical setting. PMID- 14758171 TI - Up-regulation of acetylcholine receptors during subchronic infusion of pancuronium is caused by a posttranscriptional mechanism related to disuse. AB - OBJECTIVE: Contrasting with the classic theory that competitive block of the acetylcholine receptor induces up-regulation of the receptor, recent studies show that irreversible block of acetylcholine receptors with alpha-bungarotoxin decreases acetylcholine receptor number within hours. This study investigated the early effects of competitive acetylcholine receptor block with the reversible, competitive muscle relaxant, pancuronium. DESIGN: Prospective, randomized, placebo-controlled experimental study. SUBJECTS: Healthy adult Sprague-Dawley rats. SETTING: Animal laboratory in a university hospital. INTERVENTIONS: After internal review board approval, Sprague-Dawley rats were anesthetized and received pancuronium at a rate to completely suppress neuromuscular twitch. The control group received saline. Infusion times were 0, 3, 6, or 12 hrs (n = 8 per group). One sciatic nerve was stimulated to induce muscle twitch, and the other nerve remained unstimulated. Total acetylcholine receptor expression, as well as expression of messenger RNA of the five subunits, was assayed. MEASUREMENTS AND MAIN RESULTS: There were no differences in acetylcholine receptor number between groups at time points 0, 3, and 6 hrs. At 12 hrs, acetylcholine receptor numbers in both the stimulated (35.2 +/- 4.8 fmol acetylcholine receptor/mg protein) and nonstimulated (38.3 +/- 4.8) pancuronium group, as well as the nonstimulated control saline group (37.5 +/- 4.6), were significantly increased compared with stimulated controls (27.6 +/- 4.0). Pancuronium did not potentiate the acetylcholine receptor up-regulation of the nonstimulated control group at 12 hrs. There were no changes in messenger RNA expression between groups. CONCLUSIONS: Infusion of the reversible competitive inhibitor pancuronium up to 12 hrs does not reduce acetylcholine receptor number and therefore contrasts with the irreversible acetylcholine receptor blocker alpha-bungarotoxin. This study documents that 12 hrs of disuse per se leads to an increased expression of the acetylcholine receptor number by a posttranscriptional mechanism that can be prevented by nerve-evoked muscle contraction. PMID- 14758172 TI - Neutrophil depletion attenuates interleukin-8 production in mild-overstretch ventilated normal rabbit lung. AB - OBJECTIVE: Acute lung injury induced by lung overstretch is associated with neutrophil influx, but the pathogenic role of neutrophils in overstretch-induced lung injury remains unclear. DESIGN: To assess the contribution of neutrophils, we compared the effects of noninjurious large tidal volume (Vt) ventilation on lungs in normal and neutrophil-depleted animals. SETTING: Research animal laboratory. SUBJECTS: Twenty-six male Japanese white rabbits. INTERVENTIONS: Animals were mechanically ventilated for 4 hrs with one of the three following protocols: large Vt (20 mL/kg), small Vt (8 mL/kg), and large Vt (20 mL/kg) with neutrophil depletion achieved by a single dose of vinblastine injection (0.75 mg/kg) intravenously 4 days before the experiment. MEASUREMENTS AND MAIN RESULTS: Large Vt ventilation produced alveolar neutrophil influx compared with low Vt (p =.002) without evidence of edema or increased epithelial permeability. The neutrophil influx was accompanied by increases in interleukin-8 in bronchoalveolar lavage fluid (p =.04). Immunohistochemistry of large Vt lungs showed increased interleukin-8 staining in bronchial epithelial cells, alveolar epithelium, alveolar macrophages, and smooth muscles of pulmonary vessels. Neutrophil depletion attenuated the interleukin-8 increase in the lung. Large Vt did not increase plasma interleukin-8 or tumor necrosis factor-alpha in plasma and bronchoalveolar lavage fluid. No expression of p-selectin or intercellular adhesion molecule-1 was observed. CONCLUSIONS: Cyclic overstretching of normal rabbit lungs with noninjurious large Vt produced neutrophil influx and interleukin-8 increase in bronchoalveolar lavage fluid. Production of pulmonary interleukin-8 by lung overstretch might require the interaction between resident lung cells and migrated neutrophils. This study suggests that large Vt ventilation potentiates the predisposed, subclinical lung injury, such as nosocomial pneumonia or aspiration of gastric contents. PMID- 14758173 TI - Utility of activated partial thromboplastin time waveform analysis for identification of sepsis and overt disseminated intravascular coagulation in patients admitted to a surgical intensive care unit. AB - OBJECTIVE: An abnormality of the optical transmission waveform obtained during measurement of the activated partial thromboplastin time (aPTT) has been described in association with overt disseminated intravascular coagulation. This abnormality, a biphasic waveform, is caused by the in vitro formation of Ca2+ induced complexes between very low density lipoprotein and C-reactive protein. We have evaluated the diagnostic utility of aPTT waveform analysis for identifying patients with overt disseminated intravascular coagulation and sepsis. DESIGN: Observational study investigating the predictive value of biphasic waveform for the diagnosis of sepsis and overt disseminated intravascular coagulation. SETTING: Surgical intensive care unit of a university hospital. SUBJECTS: We studied 331 consecutive patients admitted to the intensive care unit during a period of 6 months. INTERVENTIONS: Laboratory analyses, including prothrombin time, aPTT, aPTT waveform analysis, fibrinogen, D-dimer antigen, and platelet count. MEASUREMENTS AND MAIN RESULTS: At the most sensitive threshold value of the waveform variable for detection of the biphasic waveform (slope_1 = -0.05 %T/sec), this abnormality was detected in 54 of 331 patients (16.3%) at admission and 95 of 331 patients (28.7%) during the entire course of intensive care unit treatment. At this threshold, 59.3% of patients with a biphasic waveform on admission and 45.3% with a biphasic waveform during the total intensive care unit course were diagnosed with sepsis. Depending on the threshold value of slope_1, the sensitivity of aPTT waveform analysis for detection of sepsis varied between 22% and 55% at admission and between 48% and 74% during the entire intensive care unit stay. The specificity for sepsis varied between 92% and 98% and between 81% and 94%, for admission and total intensive care unit course, respectively. Biphasic waveform showed a comparable specificity for the diagnosis of overt disseminated intravascular coagulation, albeit at a lower sensitivity. CONCLUSIONS: As an adjunct to routine coagulation testing in intensive care unit patients, aPTT waveform analysis is an elegant means for the rapid and highly specific identification of patients with sepsis. PMID- 14758174 TI - Systemic, pulmonary, and hepatosplanchnic effects of N-acetylcysteine during long term porcine endotoxemia. AB - OBJECTIVE: Controversial data have been reported on the effects of N acetylcysteine in patients with septic shock. We therefore investigated the systemic, pulmonary, and hepatosplanchnic hemodynamic, gas exchange, and metabolic effects of N-acetylcysteine during long-term, volume-resuscitated, hyperdynamic porcine endotoxemia, which mimics the features of hyperdynamic human sepsis. DESIGN: Prospective, randomized, controlled experimental study. SETTING: Investigational animal laboratory. SUBJECTS: Eighteen pigs were randomized to receive endotoxin alone (controls, n = 9) or endotoxin plus N-acetylcysteine (n = 9). INTERVENTIONS: Anesthetized, mechanically ventilated, and instrumented animals received continuous intravenous endotoxin and were resuscitated with hydroxyethylstarch to keep mean arterial pressure >60 mm Hg. After 12 hrs of endotoxemia, they were randomized to receive either placebo or N-acetylcysteine (150 mg/kg loading dose over 1 hr followed by 20 mg.kg-1.hr-1 for 11 hrs). MEASUREMENTS AND MAIN RESULTS: Before as well as 12, 18, and 24 hrs after starting the endotoxin infusion, systemic, pulmonary, and hepatosplanchnic hemodynamics, oxygen exchange, and metabolism as well as nitric oxide, glutathione, and 8-isoprostane concentrations were assessed. N-acetylcysteine failed to improve any of the variables of the systemic, pulmonary, or hepatosplanchnic hemodynamics, gas exchange, and metabolism. Although N acetylcysteine significantly elevated glutathione concentration, it did not influence the 8-isoprostane concentrations and even further reduced hepatic venous pH. CONCLUSIONS: Despite the increased glutathione concentration, N acetylcysteine did not improve systemic, pulmonary, and hepatosplanchnic hemodynamics, oxygen exchange, and metabolism. When compared with previous reports in the literature, a different timing of N-acetylcysteine administration and/or an ongoing or even N-acetylcysteine-induced aggravation of oxidative stress may account for this result. PMID- 14758175 TI - Mesenteric lymph from burned rats induces endothelial cell injury and activates neutrophils. AB - OBJECTIVE: Our previous studies indicated that mesenteric lymph duct ligation prevented burn-induced lung injury. Thus, the goal of the present study was to begin to investigate potential mechanisms of this protective effect. DESIGN: Prospective animal study with concurrent control. SETTING: Small animal laboratory. SUBJECTS: Adult male Sprague-Dawley rats. INTERVENTIONS: Mesenteric lymph and portal vein plasma were collected from male rats subjected to a 40% third-degree scald burn or sham burn. The biological effects of these lymph and plasma samples were tested for their ability to kill human umbilical vein endothelial cells (HUVECs), increase HUVEC monolayer permeability, and activate polymorphonuclear leukocytes (PMNs), as reflected in CD11b adhesion molecule expression and superoxide production. Additionally, ileal specimens were harvested at the end of the experiment (6 hrs postburn) for histologic analysis. MEASUREMENTS AND MAIN RESULTS: Postburn mesenteric lymph produced during the first 2 hrs after burn injury and tested at a 5% concentration, but not sham-burn lymph or portal plasma from burned rats, was toxic for HUVECs resulting in cell death after an 18-hr incubation period. Similarly, only postburn lymph increased HUVEC monolayer permeability. Postburn lymph activated both rat and human PMNs as reflected in increased CD11b expression and augmentation of the phorbol myristate acetate-induced superoxide response. Neither sham-burn lymph nor postburn portal vein plasma activated PMNs. Both the burn and sham-burn lymph samples were sterile, indicating that the effects of burn lymph on the HUVECs or PMNs were not due to translocating bacteria. Last, an association was found between burn induced gut injury and the production of toxic burn lymph. CONCLUSIONS: Burn induced gut injury results in the production of biologically active factors that are carried in the mesenteric lymph, but not the portal plasma, which injure endothelial cells and activate PMNs and thus could contribute to distant organ injury. PMID- 14758177 TI - Peptidoglycan of Staphylococcus aureus causes inflammation and organ injury in the rat. AB - OBJECTIVE: Previous studies have implicated a role of peptidoglycan in the pathophysiology of organ injury in sepsis. However, the systemic response to, and organ injury caused by, peptidoglycan have been scarcely studied in vivo. DESIGN: Prospective, randomized study. SETTING: University-based research laboratory. SUBJECTS: Fifty-seven anesthetized, male Wistar rats. INTERVENTIONS: After surgical preparation, anaesthetized rats were administered 3 mg/kg Staphylococcus aureus peptidoglycan (n = 9), 10 mg/kg S. aureus peptidoglycan (n = 14), or an equal volume of saline (sham, n = 12) in the jugular vein over a 10-min period. MEASUREMENTS AND MAIN RESULTS: Injection of low-dose peptidoglycan (3 mg/kg) had no measurable effects on the rats. In contrast, high-dose peptidoglycan (10 mg/kg) caused increased serum values of aspartate aminotransferase (p < or =.005), alanine aminotransferase (p < or =.001), gamma-glutamyltransferase, and bilirubin (p < or =.05) (indicators of liver injury/dysfunction) as well as a moderate, but significant, increase in serum creatinine and urea (p < or =.05) (indicators of renal dysfunction). Plasma analyses showed a substantial increase in plasma values of tumor necrosis factor-alpha, interleukin-6, and interleukin 10 (p < or =.05 for all vs. sham) at 1 and 3 hrs (enzyme-linked immunosorbent assay). This was accompanied by accumulation of messenger RNAs for tumor necrosis factor-alpha, interleukin-6, and interleukin-10 in both the liver and the lung (p < or =.05 for all cytokines vs. sham) (real-time polymerase chain reaction). Peptidoglycan also caused increased DNA binding of nuclear factor-kappaB (band shift assays) and phosphorylation of c-Jun and Jun N-terminal kinase (Western blots). In the kidney, interleukin-6 messenger RNA was increased, whereas Toll like receptor 4 messenger RNA was significantly decreased. CONCLUSIONS: These results demonstrate that injection of peptidoglycan alone causes organ injury/dysfunction, organ inflammation, and systemic inflammation in the rat, involving nuclear factor-kappaB and possibly activator protein 1. These data support the contention that peptidoglycan is a contributing factor in the pathophysiology of organ injury in sepsis. PMID- 14758176 TI - Hemoglobin-vesicles suspended in recombinant human serum albumin for resuscitation from hemorrhagic shock in anesthetized rats. AB - OBJECTIVE: Hemoglobin-vesicle (HbV) has been developed to provide oxygen-carrying ability to plasma expanders. Its ability to restore the systemic condition after hemorrhagic shock was evaluated in anesthetized Wistar rats for 6 hrs after resuscitation. The HbV was suspended in 5 g/dL recombinant human serum albumin (HbV/rHSA) at an Hb concentration of 8.6 g/dL. DESIGN: Prospective, randomized, controlled trial. SETTING: Department of Surgery, School of Medicine, Keio University. SUBJECTS: Forty male Wistar rats. INTERVENTIONS: The rats were anesthetized with 1.5% sevoflurane inhalation throughout the experiment. Polyethylene catheters were introduced through the right jugular vein into the right atrium for infusion and into the right common carotid artery for blood withdrawal and mean arterial pressure monitoring. MEASUREMENTS AND MAIN RESULTS: Shock was induced by 50% blood withdrawal. The rats showed hypotension (mean arterial pressure = 32 +/- 10 mm Hg) and significant metabolic acidosis and hyperventilation. After 15 mins, they received HbV/rHSA, shed autologous blood (SAB), washed homologous red blood cells (wRBC) suspended in rHSA (wRBC/rHSA, [Hb] = 8.6 g/dL), or rHSA alone. The HbV/rHSA group restored mean arterial pressure to 93 +/- 8 mm Hg at 1 hr, similar to the SAB group (92 +/- 9 mm Hg), which was significantly higher compared with the rHSA (74 +/- 9 mm Hg) and wRBC/rHSA (79 +/- 8 mm Hg) groups. There was no remarkable difference in the blood gas variables between the resuscitated groups; however, two of eight rats in the rHSA group died before 6 hrs. After 6 hrs, the rHSA group showed significant ischemic changes in the right cerebral hemisphere relating to the ligation of the right carotid artery followed by cannulation, whereas the HbV/rHSA, SAB, and wRBC/rHSA groups showed less changes. CONCLUSIONS: HbV suspended in recombinant human serum albumin provides restoration from hemorrhagic shock that is comparable with that using shed autologous blood. PMID- 14758178 TI - A lazaroid mitigates postresuscitation myocardial dysfunction. AB - OBJECTIVE: Lazaroids, a series of 21-aminosteroids, reduce free radical mediated injury after ischemia and reperfusion. We hypothesized that the lazaroid U-74389G would minimize postresuscitation myocardial dysfunction and thereby improve neurologically meaningful survival in a rodent model after resuscitation from 8 mins of ventricular fibrillation. DESIGN: Randomized, controlled laboratory study. SETTING: University-affiliated research institute. SUBJECTS: Sprague Dawley rats. INTERVENTIONS: Ventricular fibrillation was electrically induced in ten anesthetized Sprague-Dawley rats. The lazaroid agent U-74389G in a dose of 1 mg.kg-1 or its vehicle serving as a placebo was injected into the right atrium after 7 mins of untreated ventricular fibrillation. One minute after injection of the compound, precordial compression was begun together with mechanical ventilation and continued for 6 mins before attempted electrical defibrillation. MEASUREMENTS AND MAIN RESULTS: All animals were successfully resuscitated. Postresuscitation cardiac index, left ventricular end-diastolic pressure, the rate of left ventricular pressure increase measured at a left ventricular pressure of 40 mm Hg, and the maximum rate of left ventricular pressure decline were significantly less impaired in lazaroid-treated animals. This contrasted with control animals, which had significantly greater myocardial impairment, greater neurologic deficit, and lesser duration of survival. CONCLUSIONS: The lazaroid compound U-74389G, administered during cardiac arrest, mitigated postresuscitation myocardial dysfunction and improved survival. PMID- 14758179 TI - Treatment of fever in the neurologic intensive care unit with a catheter-based heat exchange system. AB - CONTEXT: Elevated temperature worsens injury in experimental focal and global ischemia and brain trauma. Fever is common in patients with acute neurologic illness and independently predicts poor outcome. Conventional means of treating fever are not very effective in this population. OBJECTIVE: To study the effectiveness of a catheter-based heat exchange system in reducing elevated temperatures in critically ill neurologic and neurosurgical patients. DESIGN, INTERVENTION, SETTING, AND POPULATION: This was a prospective randomized, non blinded trial that compared conventional treatment of fever (acetaminophen and cooling blankets) with conventional treatment plus an intravascular catheter based heat exchange system (Alsius, Irvine, CA). Patients admitted to one of 13 neurologic intensive care units in academic medical centers were eligible if they a) suffered subarachnoid hemorrhage, intracerebral hemorrhage, ischemic infarction, or traumatic brain injury; b) had a temperature >38 degrees C on two occasions or for >4 continuous hrs; and c) required central venous access. MAIN OUTCOME MEASURE: The fever burden (area under the curve >38 degrees C) for 72 hrs was compared in an intention to treat analysis. Safety of the catheter system was monitored. RESULTS: A total of 296 patients were enrolled over 20 months. Forty one percent had subarachnoid hemorrhage, 24% had traumatic brain injury, 23% had intracerebral hemorrhage, and 13% had ischemic stroke. The groups were matched in terms of age, body mass index, sex, and Glasgow Coma Scale score distribution. Fever burden was 7.92 vs. 2.87 degrees C-hrs in the conventional group and catheter groups, respectively (64% reduction, p <.01). There was no higher rate of infection or the use of sedatives, narcotics, or antibiotics in the catheter group. The catheter did not significantly increase risk to the patient beyond that of a central catheter. CONCLUSIONS: The addition of this catheter-based cooling system to conventional management significantly improves fever reduction in neurologic intensive care unit patients. PMID- 14758180 TI - Premedication for tracheal intubation: a prospective survey in 75 neonatal and pediatric intensive care units. AB - OBJECTIVE: In children, like in adults, tracheal intubation is a painful procedure that may induce hypertension, tachycardia, and other undesirable hemodynamic disorders. Although premature neonates are very sensitive to pain and vulnerable to its long-term effects, the need for sedation before tracheal intubation is still discussed in neonatal units. Our objective was to investigate the practice of premedication before tracheal intubation in neonatal and pediatric units and determine the influence of premedication on intubating conditions. DESIGN: We performed a 10-day prospective survey in 75 neonatal and pediatric intensive care units among the 98 licensed in France. A questionnaire was completed for each intubation performed in each surveyed unit. SUBJECTS: A total of 204 patients were studied: 140 neonates, 52 infants, and 12 children. MAIN RESULTS: Data on 204 tracheal intubations were collected from 223 that were performed during the study period (participation rate, 91.4%). Premedication was used before intubation for 37.1%, 67.3%, and 91.7% of neonates, infants, and children, respectively (p <.0001). In the subgroup of neonates, premedication was particularly rare for the youngest and the smallest infants. Midazolam was the principle hypnotic used in neonates, whereas propofol was mainly used in children. Opioids or muscle relaxants were used in 16.2% and 4.4% of the patients, respectively. A low success rate and a high incidence of hypoxemia and bradycardia were correlated with the inexperience of the operator. Premedication did not significantly influence either the success rate or the undesirable events associated with tracheal intubation. CONCLUSION: Use of premedication before tracheal intubation is limited in neonates and increases according to the age of the patient. Midazolam does not seem to be an accurate choice to improve intubating conditions in neonates and infants. Because tracheal intubation is a technique that requires a skill only developed by regular practice, operators who have limited experience with intubating children should be supported by senior operators. PMID- 14758181 TI - Genomic polymorphisms in sepsis. AB - OBJECTIVE: This article aims to review all relevant genetic polymorphism studies that may contribute to the pathogenesis of sepsis with emphasis on polymorphisms of the innate immunity, pro- and anti-inflammatory cytokines, and coagulation mediators. DATA SOURCE: Published articles reporting on studies of associations between genetic polymorphisms, sepsis, septic shock, and other relevant infectious disease models. DATA ANALYSIS: Research into the pathogenesis of sepsis has led to the development of many potential therapeutic strategies. Several therapeutic agents and treatment modalities have been shown to decrease mortality rates in large, prospective, and randomized clinical trials. However, although these advances have resulted in improved survival for certain patient populations, the overall mortality rate for septic patients remains high. With the rapid development of molecular and genetic techniques, substantial interests have developed in using genomic information to define disease-mediating genetic variants in sepsis. Combined with microarray technology, it is anticipated in the near future that one will be able to tailor drug selection and dosage and predict outcome by correlating genetic profile with disease presentation. Numerous genetic association studies in sepsis have already been reported and more are likely to be published. CONCLUSIONS: Although studies examined in this review are of small heterogeneous populations, the identification of strong associations between certain genetic polymorphisms and increased mortality rate or susceptibility to severe sepsis is intriguing and supports further research using this approach. The establishment of these associations does not equal causation, and further research is required in both genetic and molecular aspect of sepsis. PMID- 14758182 TI - Airway obstruction and acute respiratory failure due to Aspergillus tracheobronchitis. AB - OBJECTIVE: To report a patient with lymphoma who developed Aspergillus tracheobronchitis resulting in airway obstruction and acute respiratory failure. DESIGN: Case report. SETTING: Intensive care unit of a tertiary care hospital. PATIENT: A 22-yr-old female with lymphoma who developed a respiratory infection 3 months after completing immunosuppressive therapy. She was treated empirically with broad spectrum antibiotics and subsequently received a supplementary chemotherapeutic course. Soon afterward she developed severe respiratory failure. Chest radiograph showed atelectasis of the right upper and lower lobes. INTERVENTIONS: Emergent mechanical ventilation; fiberoptic bronchoscopy. MEASUREMENTS AND MAIN RESULTS: Fiberoptic bronchoscopy revealed extensive obstruction of both main and subsegmental bronchi with a solid mass strongly adhered to the bronchial wall; both histologic examination and culture of that mass revealed Aspergillus. The patient died of refractory hypoxemia a few days later. CONCLUSIONS: Aspergillus tracheobronchitis should be considered in immunocompromised patients with suspected lung infection even when the main radiographic finding is atelectasis. Bronchoscopy and histologic examination of identified intraluminal material should be performed as soon as possible. PMID- 14758183 TI - Why does acute lung injury have no impact on mortality in patients with major trauma? PMID- 14758184 TI - Should we continue to target the platelet-activating factor pathway in septic patients? PMID- 14758185 TI - Therapeutic benefits of antioxidants during sepsis: is protection against oxidant mediated tissue damage only half the story? PMID- 14758186 TI - Risk of bloodstream infection can be strongly decreased by a very moderate caloric intake or strongly increased by a very low caloric intake in severely ill patients in intensive care? PMID- 14758187 TI - Do as we say, not as we do: healthcare workers and hand hygiene. PMID- 14758188 TI - Efficacy of blood transfusion in the critically ill: does age of blood make a difference? PMID- 14758189 TI - Unraveling the mystery of adrenal failure in the critically ill. PMID- 14758190 TI - Limiting life support: a world-wide consensus? PMID- 14758191 TI - Pain-related behaviors and pain assessment. PMID- 14758192 TI - Should red cell transfusions be leukoreduced in critically ill patients? PMID- 14758193 TI - Diagnosing shock via artificial intelligence: applying machine learning techniques to medicine. PMID- 14758194 TI - Peroxisome proliferator-activated receptor-gamma ligands: a pluripotent class of pharmacological agents that may prove to be useful for adjuvant treatment of sepsis and multiple organ dysfunction syndrome. PMID- 14758195 TI - Hepatic microcirculation in ischemia/reperfusion: is there a role for poly(adenosine 5'-diphosphate-ribose) polymerases? PMID- 14758196 TI - Mitochondrial membrane dysfunction in endotoxemia: the difference between what's in and what's out. PMID- 14758197 TI - Sphingomyelinase pathway activation in septic myocardial depression--another brick in the wall? PMID- 14758198 TI - Antioxidant therapy in critical illness. PMID- 14758199 TI - A new red blood cell substitute. PMID- 14758200 TI - Peptidoglycan is an important pathogenic factor of the inflammatory response in sepsis. PMID- 14758201 TI - Tracheal intubation in neonates, infants, and children: is there a right way? PMID- 14758202 TI - Antibody to enterobacterial common antigen and gram-negative sepsis. PMID- 14758203 TI - Only activated protein C treatment and not protein C has demonstrated an improvement in survival in severe sepsis. PMID- 14758205 TI - Pharyngeal flap surgery: long-term outcomes at the University of Iowa. AB - The pharyngeal flap is the most often used surgical approach to treat the problem of velopharyngeal insufficiency, a common challenge encountered in cleft palate and craniofacial clinics. The authors retrospectively reviewed short-term and long-term measures of children treated with the pharyngeal flap at the University of Iowa Cleft and Craniofacial Center. All patients who underwent pharyngeal flap surgery between January of 1970 and December of 2000, with at least one postoperative speech assessment between 2 and 5 years after the operation, were identified. Both hypernasality and hyponasality were evaluated on a scale from 1 to 6, with 1 indicating no involvement and 6 indicating severe effect on resonance. Velopharyngeal competence was also rated on a scale of 1 to 3, with 1 indicating competence and 3 indicating incompetence. These short-term data were then compared. The results showed that overall resonance performance continues to be adequate and may even improve as the patient continues to grow and mature. These findings support the use of the pharyngeal flap in the treatment of children with velopharyngeal insufficiency. PMID- 14758206 TI - Arterial anatomical features of the upper palpebra. AB - The arterial anatomical features of the upper palpebra were examined in both sides of seven fresh cadavers that had been systemically injected with a lead oxide/gelatin mixture. All specimens were stereoscopically radiographed for analysis of the three-dimensional structure of the arteries and were macroscopically dissected for observation of the relationships between the arteries and the other tissues. Cross-sections were prepared from one specimen and examined histologically. In all cases, there were four arterial arcades in the upper palpebra, namely, the marginal, peripheral, superficial orbital, and deep orbital arcades. Each arcade provided small vertical branches. The vertical branches coursed on both sides of the orbicularis oculi muscle and on both sides of the tarsal plate. From these small vertical branches, fine vessels branched off to the skin, muscle, and tarsal plate. These findings are important for avoiding complications such as bleeding and are useful for designing local flaps, such as switch flaps, for reconstructive surgical procedures. PMID- 14758207 TI - The zygomatic flap: a further possibility in reconstructing soft-tissue defects of the nose and upper lip. AB - Through the dissection and localization of the cutaneous zygomatic branch, as previously described by the authors, a vessel is available that plays an important role in reconstructive surgery. The performance of this anatomical study has enabled designing of the so-called zygomatic flap, which can be considered as a further possibility in the reconstruction of soft-tissue defects of the upper lip and nose. This new island axial pattern flap provides a reliable source of skin, with color match for facial resurfacing, and leaves a well-hidden donor site similar to that of the nasolabial flap. The flap must be carefully raised, and when properly designed, it can follow naturally existing contour lines, thus respecting and preserving the normal facial topography and leaving the patient with minimal surgical deformity. In this article, the authors report the clinical application of the zygomatic flap and the outcome of 10 cases. In a follow-up period from 1998 to the end of 2002, there was no flap loss, and in all cases, the aesthetic results were excellent and highly acceptable to the patients. The authors' experience with this new island axial pattern flap has been good, and they recommend this technique. PMID- 14758208 TI - Reconstruction of concomitant lip and cheek through-and-through defects with combined free flap and an advancement flap from the remaining lip. AB - Massive facial defects involving the oral sphincter are challenging to the reconstructive surgeon. This study presents the authors' approach to simultaneous reconstruction of complex defects with an advancement flap from the remaining lip and free flaps. From January of 1997 to December of 2001, 22 patients were studied following ablative oral cancer surgery. Their ages ranged from 32 to 66 years. Nineteen patients had buccal cancer, two patients had tongue cancer, and one patient had lip cancer. In all cases, the disease was advanced squamous cell carcinoma. Nine patients underwent composite resection of tumor with segmental mandibulectomy, and seven patients underwent marginal mandibulectomy. Cheek defects ranged from 15 x 12 cm to 4 x 3 cm, and intraoral defects ranged from 14 x 8 cm to 5 x 4 cm in size. One third of the lower lip was excised in nine patients, both the upper and lower lips were excised in 10 patients, and only commissure defects were excised in three patients. An advancement flap from the remaining upper lip was used for reconstruction of the oral commissure and oral sphincter. Then, the composite through-and-through defect of the cheek was reconstructed with radial forearm flaps in 13 patients, fibula osteocutaneous flaps in five patients, double flaps in three patients, and an anterolateral thigh flap in one patient. The free flap survival rate was 96 percent, and only one flap failed. With regard to complications, there were two patients with cheek hematoma, six patients with orocutaneous fistula or neck infection, and one patient with osteomyelitis of the mandible. All but one patient had adequate oral competence. All patients had an adequate oral stoma and could eat a regular or soft diet; two patients could eat only a liquid diet. For moderate lip defects, immediate reconstruction of complex defects took place using an advancement flap from the remaining lip to obtain a normal and functional oral sphincter; the free flap can be used to reconstruct through-and-through defects. This simple procedure can provide patients with a useful oral stoma and acceptable cosmesis. PMID- 14758209 TI - Reconstruction of large composite oromandibulomaxillary defects with free vertical rectus abdominis myocutaneous flaps. AB - Large composite oromandibulomaxillary defects resulting from oncologic resection can be challenging to reconstruct with a single flap, and functional outcomes remain anecdotal. The purpose of this study was to evaluate the authors' surgical experience and scientifically analyze and describe the functional outcomes associated with the use of the vertical rectus abdominis myocutaneous flap for reconstruction of these defects. The records of seven patients (mean age, 62 years) who underwent composite resection including hemimandibulectomy, partial maxillectomy, partial pharyngectomy, and floor-of-mouth resection followed by immediate free vertical rectus abdominis myocutaneous flap reconstruction at The University of Texas M. D. Anderson Cancer Center (1998 to 2002) were retrospectively reviewed. The tumor type was squamous cell carcinoma in all seven cases; four patients had T4 primary lesions and three had local recurrences. Radiotherapy was used preoperatively in each of the three recurrent cases (mean dose, 70.6 Gy) and postoperatively in three of the four patients with primary tumors (mean dose, 63.0 Gy). The mean length of hospitalization was 8.7 days. There were no major flap complications, fistulas, or donor-site complications. Partial flap necrosis (4 percent of flap area) occurred in one patient and dehiscence of the neck incision occurred in another. Both cases were managed with surgical debridement and closure. A third patient developed a 0.75-cm superficial suture line abscess that healed with dressing changes. The mean postoperative follow-up was 15 months. Six of the seven patients remained tube dependent for their nutrition despite some swallowing improvement; one patient returned to full oral intake. The most common swallowing deficit was impaired laryngeal excursion, which occurred in all six patients evaluated with videofluoroscopic examination and resulted in risk for aspiration in patients and frank aspiration in 83 percent. Speech was intelligible on routine follow-up visits in all patients except one. Four patients died as a result of their cancer, one was alive with metastatic disease, and two were alive with no evidence of disease at last follow up. The goal for patients undergoing extensive composite oromandibulomaxillary resection for advanced cancer is to restore structure, minimize postoperative morbidity, and optimize the quality of remaining life. Reconstruction with the free vertical rectus abdominis myocutaneous flap achieves early wound healing, allows timely delivery of adjuvant therapy, and can be accomplished with predictable success and minimal morbidity. To our knowledge, this study represents the first to scientifically analyze and quantify swallowing function following free vertical rectus abdominis myocutaneous flap reconstruction for large oromandibulomaxillary defects. Understanding of the specific physiologic swallowing deficits that typically occur after such reconstructions will provide clinicians with important surgical and reconstructive information to enable future improvements in functional success in a population for whom the prognosis is poor and treatment options are limited. PMID- 14758210 TI - Mammaplasty combining vertical and transverse approaches through a vertical incision. AB - An adjustable vertical marking is described for vertical mammaplasty in mild and moderate hypertrophy or ptosis of the breast. A vertical rectangular flap with the pedicle supported at the inframammary fold provides fullness for the upper or the lower pole of the breast. It is fixed over the pectoralis aponeurosis along the upper pole to the base of the pedicle. Length, width, and thickness of the vertical rectangular flap change regarding the extent of breast ptosis and hypertrophy. Two transverse triangular flaps, dissected in the lower pole of the breast, are supported on the inferior half of the vertical pillars at the incision margins. The criss-crossing of the triangular flaps creates a transverse support sling, avoiding the downward displacement of the breast. The vertical flap is applied in conjunction with the triangular flap to attempt to achieve projection and support for the breast with long-term stabilization of the mammary cone. Resection of mammary tissue is performed transversely just above the pedicle of the vertical flap. PMID- 14758211 TI - Importance of lateral row perforator vessels in deep inferior epigastric perforator flap harvesting. AB - Free flaps based on perforator vessels, and in particular the deep inferior epigastric perforator (DIEP) flap, are currently being applied in abdominal reconstruction. However, one of the main disadvantages is the operative complexity. Through anatomical study and clinical experience with the DIEP flap in breast reconstruction, the intramuscular path of the perforator vessels was comparatively studied, to establish the main anatomical parameters that favor procedure planning. Thirty DIEP flaps from 15 fresh cadavers were used. The number, location, and intramuscular course of the perforator vessels were determined. In addition, an initial clinical study was performed in 31 patients using 35 DIEP flaps in breast reconstruction. The number, location, and the intramuscular course of the perforators were assessed. In the cadaver study, 191 perforator vessels were detected (6.4 vessels per flap). Thirty-four percent were located in the lateral row, and the rectilinear course was observed in 79.2 percent of these vessels. In the medial row, only 18.2 percent of the perforator vessels presented this configuration (p = 0.001). Thirty-one patients underwent DIEP flap breast reconstruction, with 26 immediate and four bilateral reconstructions. In 22 of 35 flaps (62.9 percent), two perforators were used. In 25 flaps (71.4 percent), the lateral row perforators with a rectilinear course were observed. Mean operative time was 7 hours and 37 minutes. Two total flap losses and two partial necroses were observed. The majority of the lateral row perforators presented a rectilinear intramuscular course, which was shorter than that of the medial row perforators. This anatomical characteristic favors dissection with reduced operative time and vascular lesion morbidity, resulting in an important anatomical parameter for DIEP flap harvesting. PMID- 14758212 TI - Development of new reconstructive techniques: use of Integra in combination with fibrin glue and negative-pressure therapy for reconstruction of acute and chronic wounds. AB - Large wounds resulting from severe injuries are generally treated with extended reconstructive operations (e.g., free flaps), which are accompanied by long hospitalizations and risks of infection, thrombosis, and flap loss. Integra is a collagen template that can be used for reconstruction of defects. The take rate and the rate of infection are essential for the successful use of Integra (Johnson and Johnson, Hamburg, Germany). Whether the take rate and integration of Integra could be improved with the use of fibrin glue and negative-pressure therapy was assessed. Between January of 2002 and December of 2002, patients with large defects who underwent Integra grafting for reconstruction were randomly divided into groups receiving either a new treatment with fibrin glue-anchored Integra and postoperative negative-pressure therapy or conventional treatment. Demographic features, cause of the wound, location of the wound, take rate, complications of Integra coverage, time from Integra coverage to skin transplantation, and functional and aesthetic results were assessed. Twelve patients (with similar group distributions with respect to sex, age, and location and cause of the injury) were included in the study. The take rate was 78 +/- 8 percent in the conventional treatment group and 98 +/- 2 percent in the fibrin/negative-pressure therapy group (p < 0.003). The mean period from Integra coverage to skin transplantation was 24 +/- 3 days in the conventional treatment group but only 10 +/- 1 days in the fibrin/negative-pressure therapy group (p < 0.002). The decrease in the interval between coverage with Integra and skin transplantation resulted in shorter hospital stays. The use of fibrin glue and negative-pressure therapy in combination with Integra could shorten the period from coverage to integration, which would be beneficial in terms of decreased risks of infection, thrombosis, and catabolism. Therefore, it is suggested that Integra be used in combination with fibrin glue and negative-pressure therapy to improve clinical outcomes and shorten hospital stays, with decreased risks of accompanying complications. PMID- 14758213 TI - Salvage of functional elbow range of motion in complex open injuries using a sensate transposition lateral arm flap. AB - Complex open posterior elbow injuries pose three principal challenges to the reconstructive surgeon. First, the surgeon must provide stable soft-tissue closure over the joint/skeletal reconstruction. Second, the coverage must be thin and supple and promote the free gliding of the underlying structures. Finally, secondary and tertiary procedures must be anticipated beneath the flap, because a stiff, scarred, and adherent flap will only compromise these procedures. The results of 10 consecutive fasciocutaneous transposition lateral arm flaps for coverage of posterior elbow wounds are reported. This flap provides coverage that is thin and supple and that allows subsequent elevation for secondary procedures. Functionally, these flaps allowed for the development of an average arc of motion of 20 to 114 degrees and an average pronation-supination motion of 119 degrees. PMID- 14758214 TI - Pulp nonfiction: microscopic anatomy of the digital pulp space. AB - The volar pad of the fingertip provides a very stable yet sensitive surface that gives the hand the ability to pinch and grasp. The focus of this study was to advance understanding of the anatomical features of the digital pulp space. The unusual features of the fingertip pulp space include prominent collagen fiber cords and a branching continuous fine vasculature. Prominent collagen fiber cords radiating out from beneath the epidermal basement membrane are like the cords of a parachute, which directly attach to the periosteum of the distal phalanx. Those collagen fiber cords are responsible for the firm attachment of the fingertip to the distal phalanx. There is a fine patent vasculature within the pulp space. Also contained in the capsule are numerous lobules of fat, which contribute to some elasticity of the fingertip. Principles of treatment for injuries or infections of the digital pulp should attempt to preserve this anatomical construct so that the firmness and vascular supply of the fingertip are maintained and not disrupted. PMID- 14758215 TI - The surgical treatment of Dupuytren's contracture: a synthesis of techniques. AB - Dupuytren's disease is an affliction of the palmar fascia. Selective fasciectomy is recommended once contracture has occurred. Alternatives for wound closure include tissue rearrangement, the open palm technique, and full-thickness skin grafting. In this prospective study, a new "synthesis" technique was used to treat a cohort of patients with advanced Dupuytren's disease. The results were then compared with those of a second cohort of patients who underwent the open palm technique. Thirty consecutive patients were selected. Ten patients (nine men and one woman; average age, 67 years) underwent the open palm technique, and 20 patients (18 men and two women; average age, 70 years) underwent the synthesis method. Follow-up was 3.5 years for the open palm group and 2.7 years for the synthesis group. All patients in both groups improved with respect to motion, function, appearance, and satisfaction. Objectively, for the open palm technique, metacarpophalangeal joint contracture decreased from 50 degrees to 0 degrees, and proximal interphalangeal joint contracture decreased from 40 degrees to 6 degrees. Using the synthesis method, metacarpophalangeal joint contracture decreased from 57 degrees to 0 degrees, and proximal interphalangeal joint contracture decreased from 58 degrees to 10 degrees. The Disabilities of the Arm, Shoulder, and Hand Test scores decreased from 37 to 30 in both groups. There were no significant differences between groups in these parameters. The two significant intergroup differences were healing time (40 days for the open palm technique versus 28 days for the synthesis method) and recurrence rate (50 percent for open palm versus 0 percent for synthesis). The synthesis technique combines with success the best features of current methods for the surgical treatment of advanced Dupuytren's disease. PMID- 14758216 TI - Tendofascial island flap based on distal perforators of the radial artery: anatomical and clinical approach. AB - The possibility of transferring vascularized tissue to restore function and to resurface large defects, together with the use of composite flaps, has led to recent advances in "one-stage" reconstructive surgical procedures. On the basis of a previous study of the blood supply of the adipofascial flap and a new study of the blood supply of the flexor carpi radialis tendon from the transfascial and direct branches of the radial artery, a fascial island flap complete with tendon was devised and used to treat four male patients who had sustained traumatic soft tissue losses on the dorsum of the hand and segmental losses of the extensor digitorum communis. The use of a completely vascularized, single-stage, composite flap did not involve sacrifice of the radial artery, the functional and aesthetic results were good, and there was minimal donor-site morbidity. PMID- 14758217 TI - A prospective study to assess the outcome of steroid injections and wrist splinting for the treatment of carpal tunnel syndrome. AB - Surgery is the definitive treatment for carpal tunnel syndrome. Conservative treatments, such as wrist splinting and steroid injections, are also effective for the relief of carpal tunnel symptoms, but their use remains controversial because they only offer long-term relief in a minority of patients. A prospective study was performed to assess the role of steroid injections combined with wrist splinting for the management of carpal tunnel syndrome. A total of 73 patients with 99 affected hands were studied. Patients presenting with known medical causes or muscle wasting were excluded. Diagnosis was made clinically and electrodiagnostic studies were performed only when equivocal clinical signs were present. Each patient received up to three betamethasone injections into the carpal tunnel and wore a neutral-position wrist splint continuously for 9 weeks. After that period, symptomatic patients received an open carpal tunnel release, and those who remained asymptomatic were followed up regularly for at least 1 year. Patients who relapsed were scheduled for surgery. At a minimum follow-up of 1 year, seven patients (9.6 percent) with 10 affected hands (10.1 percent) remained asymptomatic. This group had a significantly shorter duration of symptoms (2.9 months versus 8.35 months; p = 0.039, Mann-Whitney test) and significantly less sensory change (40 percent versus 72 percent; p = 0.048, Fisher's exact test) at presentation when compared with the group who had surgery. It is concluded that steroid injections and wrist splinting are effective for relief of carpal tunnel syndrome symptoms but have a long-term effect in only 10 percent of patients. Symptom duration of less than 3 months and absence of sensory impairment at presentation were predictive of a lasting response to conservative treatment. It is suggested that selected patients (i.e., with no thenar wasting or obvious underlying cause) presenting with mild to moderate carpal tunnel syndrome receive either a single steroid injection or wear a wrist splint for 3 weeks. This will allow identification of the 10 percent of patients who respond well to conservative therapy and do not need surgery. PMID- 14758218 TI - Intracranial pressure changes in craniosynostotic rabbits. AB - Cranial vault and brain deformities in individuals with craniosynostosis are thought to result, in part, from changes in intracranial pressure, but clinical findings are still inconclusive. The present study describes intracranial pressure changes in a rabbit model with naturally occurring, uncorrected coronal suture synostosis. Longitudinal and cross-sectional intracranial pressure data were collected from 241 New Zealand White rabbits, divided into four groups: normal controls (n = 81); rabbits with delayed-onset coronal suture synostosis (n = 78); rabbits with early-onset unilateral coronal suture synostosis (n = 32); and rabbits with early-onset bilateral coronal suture synostosis (n = 50). Epidural intracranial pressure measurements were obtained at 10, 25, 42, and 84 days of age using a NeuroMonitor microsensor transducer. Normal rabbits and rabbits with delayed-onset coronal suture and early-onset unilateral coronal suture synostosis showed a similar oscillating pattern of age-related changes in normal and head-down intracranial pressure from 10 to 84 days of age. In contrast, rabbits with early-onset bilateral coronal suture synostosis showed markedly elevated normal and head-down intracranial pressure levels from 10 to 25 days and showed a different pattern through 84 days. Results from one-way analysis of variance revealed significant (p < 0.01) group differences only at 25 days of age. Rabbits with early-onset bilateral coronal suture synostosis had significantly (p < 0.05) greater normal and head-down intracranial pressure (by 42 percent) than the other three groups. These results showed differing intracranial pressure compensations in rabbits with uncorrected multiple-suture synostosis compared with normal rabbits or rabbits with uncorrected single-suture synostosis, possibly through progressive cerebral atrophy and decreased intracranial volume, abnormal intracranial vascular patterns and blood volume, and/or differing cranial vault compensatory changes. PMID- 14758219 TI - Distraction osteogenesis of the porcine mandible: histomorphometric evaluation of bone. AB - Distraction osteogenesis is a technique for skeletal lengthening that exploits the body's innate capacity for bone formation in response to tension forces on the repair callus. The authors developed a distraction osteogenesis model with a semiburied device in the Yucatan minipig mandible because of similarities between human and porcine mandibular anatomy, temporomandibular function, chewing patterns, and bone turnover rates. The purpose of this study was to measure histomorphometric bone fill after different latency periods, rates of distraction, and duration of neutral fixation in the minipig mandible. In addition, the relationship between histomorphometric bone fill and clinical stability was investigated. Mandibular osteotomies in 20 female Yucatan minipigs weighing 25 to 30 kg were distracted with modified semiburied distraction devices. Variables included 0-day or 4-day latency; 1-mm, 2-mm, or 4-mm daily distraction rates; gap size of 7 or 12 mm; and evaluation after neutral fixation for various lengths of time. Specimens were fixed in 2% paraformaldehyde, pH 7.4, before being embedded in methylmethacrylate. Sections were prepared from the region just below the inferior alveolar canal. The area of new bone formation within the gap was measured and expressed as a percentage of the total area of the distraction gap. Bone fill ranged from 0 to 100 percent. A pilot study with 7 mm advancements showed similar bone fill with 0-day or 4-day latency, but with poor reproducibility. Mandibles that were distracted to 12 mm at 1 mm per day exhibited nearly complete bone fill, either with 0-day latency (average, 93 percent) or 4-day latency (average, 100 percent). Mandibles that had been distracted for 3 days at 4 mm per day showed moderate osteogenesis and clinical stability with increasing time of neutral fixation. Bone fill was significantly correlated with clinical stability (Spearman r = 0.801, p = 0.001). Histological examination showed exuberant periosteal osteogenesis in distracted mandibles, even in those that showed poor bone fill and clinical stability. Thus, the periosteum appears to be a major source of new bone formation. These results show that osteogenesis was nearly complete with 1 mm per day and 0-day or 4-day latency. These results are consistent with the authors' previously reported clinical and radiographic observations that a latency period is not necessary for successful healing of the mandibular distraction osteogenesis wound. PMID- 14758220 TI - Osteocutaneous flap prefabrication in rats. AB - Composite tissue defects may involve skin, mucosa, muscle, and bone together or in combinations of two or three of these tissues. Defects involving bone and skin are frequently encountered. Osteocutaneous flaps may be used to reconstruct these composite tissue defects. Sometimes, it is not possible to obtain a vascular osteocutaneous flap. Another way of producing an osteocutaneous flap that has the desired feature is prefabrication. Prefabrication of osteocutaneous flaps can be performed in two ways: (1) a vascularized osseous flap may be grafted with skin and (2) an osteocutaneous flap can be prefabricated by implanting an osseous graft into an axial island flap. There are many articles describing osteocutaneous flap prefabrication, but there is no comparison of both methods in the literature. As an experimental model for osteocutaneous flap prefabrication, rat tail bone was chosen. For the experiments, five groups were formed. Each group contained 10 rats. In the first experimental group, a vascularized osseous segment was skin grafted and an osteocutaneous flap was prefabricated. In the second experimental group, an osseous graft was implanted into an axial skin flap. To compare viability of skin and bone components of the two prefabrication groups, vascularized tail bone was elevated with overlying skin in the third group, a bone flap was elevated in the fourth group, and a skin flap that had been prefabricated by using vascular implantation was elevated in the fifth group. The authors examined five rats in each group by microangiography at the end of 4 weeks. On microangiographic analysis, all groups showed patency of vascular pedicles. There was no difference among the groups from the point of view of vascular patency and bone appearance. Bone scintigraphy was performed on the five rats in each group. On bone scintigraphic scans, the bone component of flaps was visualized in all groups except for group 5. The mean radioactivity value on the flap side was 10,362 +/- 541.1 in group 1, 10,241 +/- 1173 in group 2, 10,696 +/- 647.1 in group 3, and 10,696 +/- 647.1 in group 4. When the radioactivity values on the flap side were compared, no statistically significant difference among groups was seen, except for group 5 (p < 0.05). To evaluate bone metabolic activity, the bone component of flap and remaining last tail bone was harvested and the radioactivity of each specimen was measured with a well-type gamma counter. The parameter of percentage radioactivity in counts per minute per unit per gram of tissue was calculated. The value of the bone component of the flap side and the value of normal bone were estimated and results were compared. The mean result was 0.86 +/- 0.08 in group 1, 0.88 +/- 0.07 in group 2, 0.87 +/- 0.07 in group 3, and 0.81 +/- 0.04 in group 4. The difference among all groups was not statistically significant. Histologic examination was performed on all rats in each group and demonstrated that the bony component was viable, showing a cellular bone marrow, osteoblasts along bony trabeculae, and vascular channels in bone-containing groups. There were no significant microangiographic, histologic, or scintigraphic differences between the two experimental methods. PMID- 14758221 TI - Chondrogenic potential of multipotential cells from human adipose tissue. AB - The use of stem cells for cell-based tissue-engineering strategies represents a promising alternative for the repair of cartilaginous defects. The multilineage potential of a population of putative mesodermal stem cells obtained from human lipoaspirates, termed processed lipoaspirate cells, was previously characterized. The chondrogenic potential of those cells was confirmed with a combination of histological and molecular approaches. Processed lipoaspirate cells under high density micromass culture conditions, supplemented with transforming growth factor-beta1, insulin, transferrin, and ascorbic acid, formed well-defined nodules within 48 hours of induction and expressed the cartilaginous markers collagen type II, chondroitin-4-sulfate, and keratan sulfate. Reverse transcription polymerase chain reaction analysis confirmed the expression of collagen type II and the cartilage-specific proteoglycan aggrecan. In summary, human adipose tissue may represent a novel plentiful source of multipotential stem cells capable of undergoing chondrogenesis in vitro. PMID- 14758222 TI - Contractile skeletal muscle tissue-engineered on an acellular scaffold. AB - For the reconstructive surgeon, tissue-engineered skeletal muscle may offer reduced donor-site morbidity and an unlimited supply of tissue. Using an acellularized mouse extensor digitorum longus muscle as a scaffold, the authors produced engineered skeletal muscle capable of generating longitudinal force. Eight extensor digitorum longus muscles from adult mice were made acellular using a protocol developed in the authors' laboratory. The acellular muscles were then placed in a bath of 20% fetal bovine serum in Dulbecco's modified Eagle's medium and 100 U/ml penicillin for 1 week at room temperature. C2C12 myoblasts were injected into the acellular muscle matrix using a 26-gauge needle and a 100 microl syringe. The resulting constructs were placed in growth medium for 1 week at 37 degrees C under 5% carbon dioxide, with media changes every 48 hours. The constructs were then placed in differentiation medium for 1 week, with media changes every 48 hours. Isometric contractile force testing of the constructs demonstrated production of longitudinal contractile force on electrical stimulation. A length-tension, or Starling, relationship was observed. Light and electron microscopy studies demonstrated recapitulation of some of the normal histologic features of developing skeletal muscle. PMID- 14758223 TI - Human embryonic kidney cells (HEK-293 cells): characterization and dose-response relationship for modulated release of nerve growth factor for nerve regeneration. AB - The development of engineered constructs to bridge nerve gaps may hold the key to improved functional outcomes in the repair of injured peripheral nerves. These constructs must be rendered bioactive by providing the growth factors required for successful peripheral nerve regeneration. Previous studies demonstrated that harvested human and rat dermal fibroblasts could be genetically engineered to release nerve growth factor (NGF) both in vitro and in vivo. The use of fibroblasts, however, has the potential to cause scarring, and the expression of NGF from those cells was transient. To overcome these potential difficulties, human embryonic kidney cells were modified for use with the ecdysone-inducible mammalian expression system. These cells (hNGF-EcR-293) have been engineered and regulated to secrete human NGF in response to the ecdysone analogue ponasterone A. HEK-293 cells were transfected with human NGF cDNA with the ecdysone-inducible mammalian expression system (Invitrogen, Carlsbad, Calif.). Stable clones were then selected. Ponasterone A, an analogue of ecdysone, was used as the inducing agent. The secretion of NGF into the medium was analyzed with two different methods. After 24 hours of exposure to the inducing agent, cell medium was transferred to PC-12 cells seeded in 12-well plates, for determination of whether the secreted NGF was bioactive. Medium from untreated or ponasterone A-treated hNGF-EcR-293 cells was deemed bioactive on the basis of its ability to induce PC 12 cell differentiation. The concentrations of secreted NGF were also quantified with an enzyme-linked immunosorbent assay, in triplicate. NGF production was measured in successive samples of the same medium during a 9-day period, with maximal release of 9.05 +/- 2.6 ng/ml at day 9. Maximal NGF production was 8.46 +/- 2.1 pg/10(3) cells at day 9. These levels were statistically significantly different from levels in noninduced samples (p 4 times control values), whereas tendon sheath fibroblasts demonstrated the highest overall levels of total TGF-beta functional activity. Lactate significantly increased TGF beta peptide (beta1, beta2, and beta3) expression, receptor (R1, R2, and R3) expression, and functional activity, suggesting a common pathway regulating tendon cell collagen production. Modulation of lactate and TGF-beta levels may provide a means of modulating the effects of TGF-beta on adhesion formation in flexor tendon wound healing. PMID- 14758226 TI - The effect of myofibroblast on contracture of hypertrophic scar. AB - Wound contraction in humans has both positive and negative effects. It is beneficial to wound healing by narrowing the wound margins, but the formation of undesirable scar contracture brings cosmetic and even functional problems. The entire mechanism of wound healing and scar contracture is not clear yet, but it is at least considered that both the fibroblasts and the myofibroblasts are responsible for contraction in healing wounds. The myofibroblast is a cell that possesses all the morphologic and biochemical characteristics of both a fibroblast and a smooth muscle cell. Normally, the myofibroblasts appear in the initial wound healing processes and generate contractile forces to pull both edges of an open wound until it disappears by apoptosis. But as an altered regulation of myofibroblast disappearance, they remain in the dermis and continuously contract the scar, eventually causing scar contracture. In this research, to compare and directly evaluate the influence on scar contracture of the myofibroblast versus the fibroblast, dermal tissues were taken from 10 patients who had highly contracted hypertrophic scars. The myofibroblasts were isolated and concentrated from the fibroblasts using the magnetic activating cell sorting column to obtain the myofibroblast group, which contained about 28 to 41 percent of the myofibroblasts, and the fibroblast group, which contained less than 0.9 percent of the myofibroblasts. Each group was cultured in the fibroblast populated collagen lattice for 13 days, and the contraction of the collagen gel was measured every other day. In addition, they were selectively treated with tranilast [N-(3',4'-dimethoxycinnamoyl) anthranilic acid] to evaluate the influence on the contraction of the collagen gel lattice. During the culture, the myofibroblast group, compared with the fibroblast group, showed statistically significant contraction of the collagen gel lattice day by day, except on the first day, and only the myofibroblast group was affected by tranilast treatment, showing significant inhibition of gel contraction. By utilizing an in vitro model, the authors have demonstrated that myofibroblasts play a more important role in the contracture of the hypertrophic scar. PMID- 14758227 TI - Deforming posttraumatic hematoma of the nasal tip: an infrequent lesion. PMID- 14758228 TI - Antegrade venous drainage in a reverse radial forearm flap. PMID- 14758229 TI - A simple method for lower lateral cartilage repositioning in cleft lip nose deformity. PMID- 14758230 TI - Design of the Limberg flap by a specially designed ruler: a personal approach. PMID- 14758231 TI - Iliac crest donor defect: avoiding the limp. PMID- 14758232 TI - The vacuum expandable condom mold: a simple vaginal stent for McIndoe-style vaginoplasty. PMID- 14758233 TI - Lay evaluations of speech results following palatoplasty revisited. PMID- 14758234 TI - Outcome assessment of an in-hospital cross-functional wound care team. PMID- 14758235 TI - Restoring abdominal wall integrity in contaminated tissue-deficient wounds using autologous fascia grafts. PMID- 14758236 TI - Congenital nasal anomalies: a classification scheme. AB - The purpose of this work was to develop a simple yet comprehensive classification scheme dedicated to congenital nasal anomalies. To date, no such classification system has been proposed and widely used. A 22-year retrospective review was performed. Two hundred sixty-one patients with congenital nasal anomalies were identified. From this extensive database, a systematic morphogenic classification system was devised. Congenital nasal deformities were classified into four categories. Type I, hypoplasia and atrophy, represents paucity, atrophy, or underdevelopments of skin, subcutaneous tissue, muscle, cartilage, and/or bone. Type II, hyperplasia and duplications, representing anomalies of excess tissue, ranging from duplications of parts to complete multiples, are categorized here. In the type III category, clefts, the comprehensive and widely utilized Tessier classification of craniofacial clefts is applied. Type IV deformities consist of neoplasms and vascular anomalies. Both benign and malignant neoplasms are found in this category. PMID- 14758237 TI - Fifty: another year older or a milestone? PMID- 14758238 TI - The anatomy of the greater occipital nerve: implications for the etiology of migraine headaches. AB - An interest in pursuing new theories of the underlying etiology of migraine headaches has been sparked by previously published reports of an association between amelioration of migraine headache symptoms and corrugator resection during endoscopic brow lift. This theory has further been reinforced by recent publications documenting improvement in migraine headaches following injection of botulinum A toxin. There are thought to be four major "trigger points" along the course of several peripheral nerves that may cause migraine headaches. Among these peripheral nerves is the greater occipital nerve. For this reason, the authors have undertaken an anatomic study of this nerve to determine its usual course, potential anatomic variations, and possible points of potential entrapment or compression. The results of this anatomic study have enhanced further development of techniques designed to address these points of entrapment/compression and potentially lead to relief of migraine headaches caused by this mechanism. Twenty cadaver heads from patients with an unknown history of migraine headaches were dissected to trace the normal course of the greater occipital nerve from the semispinalis muscle penetration to the superior nuchal line. Standardized measurements were performed on 14 specimens to determine the location of the emergence of the nerve using the midline and occipital protuberance as landmarks. On the basis of this information, the location of emergence was determined to be at a point centered approximately 3 cm below the occipital protuberance and 1.5 cm lateral to the midline. This location can, in turn, be used to guide the practitioner performing chemodenervation of the semispinalis capitis muscle in an attempt to provide migraine symptom relief. PMID- 14758239 TI - Prospective analysis of the outcome of subpectoral breast augmentation: sensory changes, muscle function, and body image. AB - This study is a prospective analysis of the outcome of subpectoral breast augmentation. Forty-seven patients undergoing breast augmentation were studied. They were assessed for pectoralis muscle function, breast sensation, and body image before and after subpectoral breast augmentation with saline implants. The patients were evaluated as follows: Pectoralis function was determined by measuring maximal voluntary isometric force. Sensation was evaluated by two means: vibration and pressure. The patient's body image was assessed using the Multidimensional Body-Self Relations Questionnaire. Results indicated a significant change in breast sensation at 3 months postoperatively but not at 6 months. Pectoralis muscle function did not significantly change during the study period. Body image was significantly improved at both postoperative measuring periods. The authors conclude that breast augmentation results in improved body image with negligible effect on muscle or nerve function. PMID- 14758240 TI - Incidence of earlobe ptosis and pseudoptosis in patients seeking facial rejuvenation surgery and effects of aging. AB - The authors have previously described a classification system for earlobe ptosis and have established a criterion for earlobe pseudoptosis. Earlobe heights were characterized based on anatomic landmarks, including the intertragal notch, the otobasion inferius (the most caudal anterior attachment of the earlobe to the cheek skin), and the subaurale (the most caudal extension of the earlobe free margin). The classification system was derived from earlobe height preferences as determined by a survey of North American Caucasians, and it identified the ideal free caudal lobule height range to measure 1 to 5 mm from otobasion inferius to subaurale (grade I ptosis). Also, earlobe pseudoptosis was defined by the attached cephalic lobule height measuring an intertragal notch to otobasion inferius distance greater than 15 mm. In this study, the preoperative earlobe height measurements of 44 patients seeking facial rejuvenation were evaluated. The average attached cephalic segment (intertragal notch to otobasion inferius distance) of patient earlobes measured 11.10 +/- 0.46 mm, and the average free caudal segment (otobasion inferius to subaurale distance) of patient earlobes measured 7.15 +/- 0.49 mm. Assessment of patient groups based on single-decade age differences demonstrated an increase in the free caudal segment (otobasion inferius to subaurale distance) with increasing age (p = 0.003). Assessment of patient groups based on single-decade age differences demonstrated no increase in the attached cephalic segment (intertragal notch to otobasion inferius distances) with increasing age (p = 0.281). When evaluating for the ideal otobasion inferius to subaurale distance, only 22.2 percent of earlobes demonstrated an ideal free caudal earlobe height (grade I ptosis). Moreover, pseudoptosis was detected in 12.3 percent of earlobes. Finally, a majority of earlobes demonstrated intrapatient variability, with only 16.2 percent of patients demonstrating identical attached cephalic segment (intertragal notch to otobasion inferius distances) and 37.8 percent demonstrating identical free caudal segment (otobasion inferius to subaurale distances) when compared with their contralateral ear. Plastic surgeons should be aware that a significant number of patients (77.8 percent of earlobes) may not possess an ideal free caudal segment and that 12.3 percent of earlobes may present with pseudoptosis. Therefore, earlobe height assessment should be an essential aspect of evaluation in patients desiring facial rejuvenation surgery. Evaluation of both ears should be performed independently due to intrapatient earlobe height variations. Finally, patients should be counseled with regard to the ideal earlobe parameters and aging patterns (stable attached cephalic segment versus increasing free caudal segment). With the natural progression of both facial rhytides and caudal segment earlobe ptosis (increasing free lobule segment) with increasing age, independent and accurate assessment of earlobe height is indicated so that the aging ear may be addressed concurrently with the aging face. PMID- 14758241 TI - Tumescent and dry liposuction of lower extremities: differences in lymph vessel injury. AB - Lipectomy is a standard procedure in plastic surgery. Until now, however, there was no definite information about the influence of different liposuction techniques (tumescent versus dry liposuction) on the integrity of lymph collectors during this procedure. To study the effect of these liposuction techniques on the incidence of lymph vessel injury, postmortem lymphatic preparations were done in nine human cadavers (18 lower extremities). Conventional liposuction with a blunt 4-mm cannula in the dry technique (n = 29 regions) was compared with the tumescent technique (n = 26). Liposuction was performed in parallel to the superficial lymph vessels (longitudinal suction) or transversally in an 80-degree to 90-degree angle to the extremity (vertical suction). Careful surgical preparation of different regions followed. A specific macroscopic lymph vessel injury score was applied to differentiate three degrees of lymph vessel lesions according to the extravasation of patent blue. In all lower extremities, postmortem lymph flow occurred as indicated by patent blue staining of the lymph vessels. Injection of fluid that is obligatory during tumescent suction did not result in grade 2 injury. On the contrary, tumescent suction overall produced significantly fewer lymph vessel lesions when compared with the dry technique (p < 0.05). Longitudinal liposuction produced significantly less injury when compared with vertical suction (p < 0.05). Tumescent suction and dry suction were equally effective in removing adipose aspirates, as verified by circumference measurements. In addition, tumescent liposuction is unlikely to cause major lesions of epifascial lymph vessels during suction procedures vertical to the extremity axis. Therefore, in this respect, this technique is superior to dry suction. PMID- 14758242 TI - Circumferential suction lipectomy of the trunk with anterior rectus fascia plication through a periumbilical incision: an alternative to conventional abdominoplasty. AB - During the past decade, many combinations of operative techniques for abdominoplasty have evolved to suit the individual requirements of the patient. The purpose of this study was to present a safe alternative to conventional abdominoplasty for appropriate patients, namely, those with minimal skin laxity, moderate fatty tissue distribution, musculofascial diastases, and no concern for abdominal stretch marks. The procedure consists of circumferential ultrasound assisted liposuction and direct abdominal wall plication through a periumbilical incision. Thirty-two patients were evaluated on overall patient satisfaction and complication rates, including seroma (five, 15.6 percent), major sensory loss (none), skin slough (none), skin burns (none), end hits (i.e., a small, partial thickness, subdermal burn; one, 3 percent), and limited results (two, 6 percent). The patients expressed that avoidance of the abdominal scar and diminished recuperative time outweighed the benefit of tighter skin associated with conventional abdominoplasty. This technique may provide another avenue for appropriate contouring of the abdomen in properly selected patients. PMID- 14758243 TI - Update on Mitek endoscopic brow fixation system. PMID- 14758244 TI - The way it really happened: Breslow, Macht, and tumor thickness in melanoma. PMID- 14758245 TI - The March mismatch: a tale of misconception and misfortune. PMID- 14758246 TI - What doctors aren't being told: using the freedom of information act. PMID- 14758247 TI - Isn't it time to remodel yourself? PMID- 14758248 TI - Safer sharps. PMID- 14758250 TI - Thoughts of a young plastic surgeon. PMID- 14758251 TI - After years in Congress, it's good to be a plastic surgeon again! PMID- 14758252 TI - The history of otolaryngology in plastic surgery. PMID- 14758253 TI - A giant sarcoma of the scalp. PMID- 14758254 TI - A case of adult T-cell leukemia/lymphoma. PMID- 14758255 TI - Treatment of coup de sabre deformity with porous polyethylene implant. PMID- 14758256 TI - Use of AlloDerm and polytetrafluoroethylene together to correct a depression of the frontal bone. PMID- 14758257 TI - A heterotopic mimic muscle positioned in the temporal region. PMID- 14758258 TI - Comments regarding orbitoblepharophimosis syndrome: a 16-year perspective. PMID- 14758259 TI - The use of suture cover as an internal nasal splint. PMID- 14758260 TI - The temporal sequence of periosteal attachment after elevation. PMID- 14758261 TI - Self-involution of giant keratoacanthoma on the tip of the nose. PMID- 14758262 TI - A painful neuroma in the nose following aesthetic rhinoplasty. PMID- 14758263 TI - Zigzag draping over the elevated helical framework. PMID- 14758264 TI - Ear reconstruction after auricular chondritis secondary to ear piercing. PMID- 14758265 TI - Repair of extensive tissue slough of the face which followed insertion of coils to control epistaxis. PMID- 14758266 TI - Perioral dermatitis following orthognathic surgery. PMID- 14758267 TI - An unusual movement of the tongue. PMID- 14758268 TI - Mohs out of control. PMID- 14758269 TI - Hypopharynx reconstruction. PMID- 14758270 TI - Palatopharyngeal lipoinjection: an advantageous method in velopharyngeal incompetence. PMID- 14758271 TI - Does fascia provide additional, meaningful coverage over a breast implant? PMID- 14758272 TI - An easy way to convey the vascular pedicle of the free flap through the tunnel. PMID- 14758273 TI - A novel technique for connecting tubing placement of remote subcutaneous tissue expander ports. PMID- 14758274 TI - Breast reconstruction with deepithelialized DIEP flap after recurrent mastitis. PMID- 14758275 TI - Pain control in augmentation mammaplasty: the use of indwelling catheters in 200 consecutive patients. PMID- 14758276 TI - Minimally invasive localization technique: needle fixation of subcutaneous lipoma. PMID- 14758277 TI - Wristwatch extensor tendon injury. PMID- 14758278 TI - Osteofasciocutaneous forearm and fibular flap donor-site morbidity revisited. PMID- 14758279 TI - Holding a cut tendon safely. PMID- 14758280 TI - The possible protective effects of antioxidants in ultrasound-assisted lipoplasty. PMID- 14758281 TI - Adipofasciocutaneous flap coverage of a fifth toe cicatrix following medicated pad treatment of a hard corn. PMID- 14758282 TI - An aid to the greater saphenous vein exclusion from the posterior tibial perforator-saphenous flap. PMID- 14758283 TI - New application of the distally based sural artery flap. PMID- 14758284 TI - Toe reconstruction following a lawnmower injury using a distally based adipofascial turnover flap and distraction osteogenesis. PMID- 14758285 TI - An accessible and simple cast cover. PMID- 14758286 TI - An interesting and useful balsamic resin used to increase the adherence of adhesive tape to the skin: Siam benjoin (tentuire de benjoin). PMID- 14758287 TI - A simplified technique for split-thickness skin graft donor-site care. PMID- 14758288 TI - Use of gallipot for debridement. PMID- 14758289 TI - Erythema annulare centrifugum: an unusual presentation for acute leukemia. PMID- 14758291 TI - When and for whom does the bell toll? PMID- 14758290 TI - Surgical management of the cutaneous manifestations of linear nevus sebaceus syndrome. PMID- 14758292 TI - Lay clinics and an epidemic outbreak of mycobacterium skin and soft-tissue infection. PMID- 14758293 TI - Trunk muscle strength in relation to balance and functional disability in unihemispheric stroke patients. AB - OBJECTIVE: To evaluate trunk muscle strength in unihemispheric stroke patients and to assess how it relates to body balance and functional disability in this patient group. DESIGN: This prospective case-comparison study investigated isometric and isokinetic reciprocal trunk flexion and extension strength at angular velocities in 38 unihemispheric stroke patients and 40 healthy volunteers. The Berg balance scale was used to assess balance and stability, and the FIM instrument was used to evaluate functional disability in the patient group. Patients were evaluated as soon as they were able to stand long enough for testing. RESULTS: Peak torque values for trunk flexion and extension were lower in the stroke patients than in the controls. The differences were significant for trunk flexion and for trunk extension. In both groups, peak torque values for trunk flexors were greater than peak torque values for trunk extensors. There was a significant positive correlation between trunk muscle strength and Berg balance scale score at discharge. Trunk muscle strength was not correlated with FIM total score or FIM motor score, but the locomotion-transfers FIM subscore at discharge was positively correlated with trunk muscle torque values, except for isometric extension. CONCLUSION: The findings indicate trunk flexion and extension muscle weakness in unihemispheric stroke patients, which can interfere with balance, stability, and functional disability. PMID- 14758294 TI - Full-time integrated treatment program, a new system for stroke rehabilitation in Japan: comparison with conventional rehabilitation. AB - OBJECTIVE: To validate the effectiveness of the Full-time Integrated Treatment (FIT) program that is characterized by rehabilitation 7 days/wk, encouragement of daytime activity, and enhanced communication between staff in stroke rehabilitation. DESIGN: Since our facility changed from the conventional rehabilitation system of 5 days of treatment to the FIT program in December 2000, we compared the conventional rehabilitation program with the FIT program at our hospital. The conventional treatment group and the FIT group consisted of 48 and 58 first-stroke hemiplegics, respectively. RESULTS: The motor subscore of the FIM instrument at admission and at discharge was 64.3 and 77.0 in the conventional group and 60.6 and 80.9 in the FIT group, respectively. The length of stay and efficiency of the FIM instrument were 80.0 days and 0.16 in the conventional group and 69.8 days and 0.30 in the FIT group, respectively. These differences between groups were statistically significant, with the exception of admission FIM data. CONCLUSIONS: Because the FIT program attained a higher discharge FIM level with a shorter length of stay, the FIT program was concluded to be an efficient and effective method of stroke rehabilitation. PMID- 14758295 TI - Effectiveness of a multidisciplinary occupational training program for chronic low back pain: a prospective cohort study. AB - OBJECTIVE: To evaluate the effectiveness of a 12-wk multidisciplinary occupational training program for patients with chronic low back pain and to identify prognostic factors for treatment success. DESIGN: A total of 51 participants were evaluated at baseline, at discharge, and at 1 yr after conclusion of the program. The evaluation included a physical examination and assessment of functional disability, psychological factors, and coping styles. The main target of the program is full work resumption. The central outcome measures therefore are three variables on return to work. RESULTS: Analysis of variance for repeated measures revealed significant beneficial changes during the program for all measures except for several coping-style variables. The acquired level of maximum oxygen uptake, trunk flexibility, functional disability, and catastrophizing were maintained at 1-yr follow-up. At 1-yr follow-up, >60% of the participants had fully returned to work, which is an increase of >40% compared with baseline. Regression analyses showed that sex, age, the baseline values of reinterpretation of pain sensations, and functional disability and changes in trunk flexibility scores during the program are important prognostic factors for complete return to work. CONCLUSIONS: Based on the current findings, the program seems to be efficacious in the short term. Future attention must be directed toward maintaining these results, although work resumption rates improved considerably 1 yr after conclusion of the program. PMID- 14758296 TI - Electrical twitch-obtaining intramuscular stimulation in lower back pain: a pilot study. AB - OBJECTIVES: To determine if electrical twitch-obtaining intramuscular stimulation (ETOIMS) provides greater myofascial lower back pain relief than muscle stimulation or skin stimulation. DESIGN: In this single-blinded, crossover, pilot trial performed at a university-affiliated outpatient rehabilitation medicine department in Taiwan, 12 acupuncture-naive patients with lower back pain of 3-60 mos duration received one crossover treatment every 2 wks by monopolar needle electrode insertion at bilateral T10-S1 levels to: (1) paraspinal muscles, (2) overlying skin, and (3) paraspinal muscles with ETOIMS applied via the needle electrode at individual treatment sites. A total of 30 manual insertions per side per treatment were performed, with withdrawal after 2 secs. Beginning 1 wk before each trial and continuing until 2 wks after, patients completed a visual analog scale twice daily. In addition, on the day of treatment, patients received a physical examination and completed a visual analog scale both before and after treatment. RESULTS: Significant and immediate reduction in the visual analog scale levels was noted only with ETOIMS. Immediate improvement occurred in one of nine physical tests with muscle stimulation and ETOIMS only. In the 2 wks after treatment, absolute visual analog scale levels for ETOIMS were significantly lower than muscle stimulation and skin stimulation. ETOIMS resulted in a greater percentage of pain relief in the first week after treatment, although it was not statistically significant from muscle stimulation and skin stimulation. CONCLUSIONS: ETOIMS provided significantly greater immediate and sustained myofascial lower back pain relief than muscle stimulation and skin stimulation. Although a greater percentage of pain reduction occurred with ETOIMS, it was not statistically significant. PMID- 14758297 TI - Simultaneous, bilateral, and three-dimensional gait analysis of elderly people without impairments. AB - OBJECTIVE: The main objectives of this study were to determine if the right and left lower limbs in the gait of elderly people developed similar muscle moment and power patterns and to determine whether their associated mechanical energies were different during two consecutive gait cycles. DESIGN: A total of 18 able bodied male subjects aged 71 +/- 6.8 yrs participated in this study. Data were collected using three Optotrak position sensors and two AMTI force platforms. RESULTS: The peak muscle powers were very similar in the sagittal plane and reflected gait symmetry, except for greater and significant differences produced by the hip extensors. Differences in the frontal and transverse planes were mainly attributed to the actions taken by the muscles, leading the lower limbs to compensate and to dynamically balance and propel the body forward during two consecutive gait cycles. In terms of mechanical energy, 13% greater positive work was done in the left limb and was associated with asymmetrical behavior of the lower limbs to propel the body forward. The total negative work was similar for both limbs, and substantial work was done in the frontal plane by the hip, indicating the role of muscle activity in this plane to control the pelvis and trunk against gravitational forces. CONCLUSIONS: In addition to the importance of a balance-control function in the gait of healthy, elderly subjects, muscle activity made an important contribution to propelling the body forward. Gait asymmetry in elderly subjects seems to be related to different degrees of energy generated by the lower limbs for the propulsion function, whereas both limbs contribute similarly to the balance-control function. PMID- 14758299 TI - Effect of methylphenidate on vital signs and adverse effects in adults with traumatic brain injury. AB - OBJECTIVE: To study methylphenidate's adverse effects and impact on vital signs within the adult traumatic brain injury population. DESIGN: Thirty-five adults with traumatic brain injury enrolled in a double-blind, placebo-controlled, 6-wk crossover study of methylphenidate, given in a dose of 0.3 mg/kg/dose, twice a day. Vital signs were taken by trained clinicians and research assistants. Participants filled out weekly questionnaires pertaining to the adverse effects. RESULTS: Poor appetite was the only adverse effect related to methylphenidate. Other adverse effects commonly associated with methylphenidate, such as insomnia, rapid heart rate, and anxiety, were not found to be significantly related to the medication. The average rise in mean arterial pressure on methylphenidate was 2.5 mm. Methylphenidate showed a stronger impact on pulse, with an average increase of 7 beats/min. Baseline vital signs did not predict the degree of increase on methylphenidate. CONCLUSION: Methylphenidate appears to be safe for the adult population with traumatic brain injury. However, because a few individuals experienced significant changes in vital signs and adverse effects, all patients should be monitored. PMID- 14758298 TI - Transmetatarsal amputation prosthesis with carbon-fiber plate: enhanced gait function. AB - OBJECTIVE: To determine whether the design of a transmetatarsal amputation prosthesis with a carbon-fiber plate would improve gait pattern in patients with transmetatarsal amputations. DESIGN: In the gait laboratory in a tertiary medical center, eight male patients with transmetatarsal amputations were recruited. Nine able-bodied male volunteers were recruited as the control group. A full-length standard shoe and a transmetatarsal amputation prosthesis with a carbon-fiber plate were the footwear used. Our transmetatarsal amputation prosthesis included a custom-molded insole, a mounted toe filler, and a thin, lightweight, carbon fiber plate incorporated directly beneath the insole. RESULTS: After wearing the transmetatarsal amputation prosthesis with a carbon-fiber plate, the results of the kinetic and kinematic studies were summarized. CONCLUSION: The transmetatarsal amputation prosthesis with a carbon-fiber plate improved gait pattern significantly in patients with transmetatarsal amputations. Drastic shoe modifications are not necessary to have the prosthesis inserted. The carbon-fiber plate functions like the spring-steel shank within the sole of the shoe. The carbon-fiber plate, the toe filler, and the total-contact insole are all mounted as a whole to ensure better foot contact. Therefore, our transmetatarsal amputation prosthesis with a carbon-fiber plate can be a good alternative choice of footwear in patients with transmetatarsal amputations. PMID- 14758300 TI - Pain management in trauma patients. AB - Trauma is a major cause of mortality throughout the world. In recent years, major advances have been made in the management of trauma, the end result of which has been reduced mortality and enhanced function. One of these areas is pain control. Improved pain management has not only led to increased comfort in trauma patients, but has also been shown to reduce morbidity and improve long-term outcomes. This review focuses on the treatment of pain in the setting of acute injury and on pain management in trauma patients who go on to develop chronic pain. Emphasis is placed on pharmacologic interventions, invasive and noninvasive pain management techniques, analgesia in challenging patients, and pain control in commonly encountered trauma conditions. PMID- 14758301 TI - "Young" cervical spinal stenotic: a review of 118 patients younger than 51 years of age. AB - A 10-yr retrospective review of 460,964 admissions to a 1,000-bed community teaching hospital identified 555 patients with a diagnosis of cervical spinal stenosis. Of this number, 118 were classified as "young." With an occurrence rate of 26 per 100,000, in this series there was not a significant difference in sex throughout the decade. During the first 5 yrs of this study, 186 patients with cervical spinal stenosis were identified, of whom 37 were <51 yrs of age. Of 369 patients in the remaining 5-yr period with cervical spinal stenosis, 81 were <51 yrs of age. The proportion of those <51 yrs old between the two 5-yr periods was not statistically different. Although spinal stenosis, both cervical and lumbar, has been regarded as a disorder of the elderly, 21% of those with cervical spinal stenosis and 10% with lumbar spinal stenosis can be anticipated to be <51 yrs of age. PMID- 14758302 TI - Pushrim-activated power-assist wheelchairs: elegance in motion. PMID- 14758303 TI - Children with cancer, fever, and treatment-induced neutropenia: risk factors associated with illness requiring the administration of critical care therapies. AB - OBJECTIVES: To identify clinical and laboratory characteristics of pediatric patients with cancer, fever, and treatment-induced neutropenia, available at existing at initial presentation, that are independently associated with the development of illnesses requiring administration of critical care therapies. METHODS: We retrospectively collected historical, clinical, and laboratory data on initial presentation for all pediatric (younger than 18 years) cancer patients admitted for fever and treatment-induced neutropenia at our institution over a 5 year period. The outcome variable was the need for administration of a critical care therapy within 24 hours of admission. A multivariable analysis was performed and internally validated using bootstrap analysis. RESULTS: We identified 303 events in 143 patients, of which 36 (11.9%) received a critical care therapy. Higher temperature at presentation and capillary filling time (CFT) of >3 seconds retained significance in the multivariable analysis and were validated by the bootstrap analysis. The positive and negative predictive values of the presence of either temperature of > or =39.5 degrees C or CFT of >3 seconds were 35% and 91%, respectively. CONCLUSIONS: Pediatric patients with cancer, fever, and treatment-induced neutropenia who present with higher fever or prolonged CFT are at increased risk of developing life-threatening illnesses requiring administration of critical care therapies, independent of hematologic factors, type of cancer, or other physiologic signs of sepsis. PMID- 14758304 TI - Clinical course of urinary tract infections in infants younger than 60 days of age. AB - OBJECTIVES: Although often managed differently than older children, no study has specifically described the clinical course of urinary tract infections (UTIs) in young infants. Our objective was to determine the risk of progression of illness and the pattern of fever resolution in infants younger than 60 days of age with Gram-negative rod UTIs. METHODS: We completed a retrospective medical chart review. Patients younger than 60 days of age presenting to an urban, tertiary care pediatric hospital were included if they had single organism growth of Gram negative rods in any amount from suprapubic aspiration samples or more than 10,000 cfu/mL from catheterized specimens. Significant progression of illness was defined as the need for transfer to an intensive care setting. Fever was defined as a rectal temperature of 38.0 degrees C or higher or an axillary temperature 37.0 degrees C or higher. Temperatures were assigned to blocks of 4 hours. RESULTS: Of 128 patients with available records, none were transferred from the general pediatric ward to the intensive care unit and 2 were transferred to a step-down unit for events potentially unrelated to the UTI. No patient had a positive repeat urine culture. For patients with fever, median time to fever resolution was within the 4 to 8 hour time block. Eighty-five percent of the febrile patients became afebrile within 24 hours and only 3.6% were febrile after 48 hours. CONCLUSIONS: Progression of illness in infants with Gram-negative rod UTIs is unlikely. Fever resolution is rapid. If subsequent studies concur with our findings, outpatient therapy or short-stay unit admission may become a viable management strategy. PMID- 14758305 TI - Underdosing of acetaminophen by parents and emergency department utilization. AB - OBJECTIVES: Fever is a common reason for parents to seek medical attention for their children. We conducted this study to document accuracy of parental administration of acetaminophen and to identify if parents who did not give an optimal dose would have decided not to come to the emergency department (ED) if the fever had diminished at home. METHODS: A cross-sectional study including 248 caregivers of children who had a chief complaint of fever and had been given acetaminophen in the preceding 24 hours were interviewed. RESULTS: Enrollment was 86%. One hundred parents (47%) gave acetaminophen in the recommended dose, 26 parents (12%) gave an overdose, and 87 (41%) gave an underdose of acetaminophen. Half of the parents (54%) would not have come to the ED if the fever had subsided after using the antipyretic treatment at home. Children with significantly higher maximal temperature at home would not have been taken to the ED if fever had subsided. Parents who speak English as the primary language at home gave the recommended dose of acetaminophen more frequently than non-English-speaking parents. CONCLUSION: A significant portion of our population gives an underdose of acetaminophen, reflecting lack of knowledge or misuse. Based on parental reports, the majority of visits for fever might have been prevented, if parents had been successful in their effort to reduce temperature to below of what they considered as fever, but factors other than underdosing of acetaminophen probably encourage parents of febrile children to visit the ED. PMID- 14758306 TI - A randomized, prospective, multisite comparison of pediatric prehospital training methods. AB - OBJECTIVE: Results of prehospital pediatric continuing education using train-the trainer and CD-ROM training methods were compared to each other and to a control group. The null hypothesis was that no differences would be found in pretraining and posttraining measurements of knowledge and performance by either training method. METHODS: This was a prospective trial involving 12 sites. Random selections were made from ambulance service lists provided by 3-state emergency medical services (EMS) agencies. Preintervention and postintervention (12-month) measurements included a written examination and 2 performance scenarios videotaped for independent panel evaluation. Training was either an interactive CD-ROM or standard classroom instruction using a train-the-trainer model. Mean differences in written, performance, and combined scores were analyzed. RESULTS: Differences were noted in the combined and performance scores for the CD-ROM intervention group. No differences were noted in written measurements between or among the groups. CONCLUSION: In this small sample, interactive CD-ROM training shows promise for improving performance. The research design, with additional guards against sample size attrition, may provide a model for multisite EMS education research. PMID- 14758307 TI - Success rates of pediatric intubation by a non-physician-staffed critical care transport service. AB - OBJECTIVES: Previous researchers have found that institution of an endotracheal intubation (ETI) protocol into a large urban paramedic program resulted in low success rates and had no beneficial effects. The primary goal of the current study was to assess ETI success rates achieved by a small cadre of nonphysician critical care transport (CCT) providers. A secondary objective was to assess for association between ETI success and factors such as age group or ETI setting (eg, in-hospital, in-aircraft). DESIGN: This retrospective study analyzed transport records of consecutive pediatric patients (younger than 13 years) in whom ETI was attempted by a nurse/paramedic (RN/EMTP) CCT crew working under protocols which included neuromuscular blockade (NMB)-facilitated ETI. The CCT service performs scene and interfacility transports in helicopter, fixed-wing (airplane), and ground critical care vehicles; pediatric patients are transferred to 4 receiving tertiary care centers. Chi2 test, Fisher exact test, and logistic regression analysis (P = 0.05) examined ETI success rates and assessed for association between ETI success and various characteristics (eg, age group, ETI setting). RESULTS: The CCT crew attempted ETI in 143 patients, with success in 136 cases (95.1%). There were no unrecognized esophageal intubations. ETI success was of similar likelihood across pediatric age groups (P = 0.19) and in different ETI settings (P = 0.57). CONCLUSIONS: CCT crew airway management success was very high in all practice settings. These data support contentions that, with a high level of initial and ongoing training, nonphysician CCT crew can successfully manage pediatric airways in a variety of circumstances. PMID- 14758308 TI - Acute presentation of infected urachal cysts: case report and review of diagnosis and therapeutic interventions. AB - Urachal remnants, although relatively rare, masquerade as a large number of diverse disorders leading to a high rate of misdiagnosis. A typical case is reported in which a 10-year-old boy presented to the Emergency Department twice before being incorrectly diagnosed with a pelvic or lower abdominal periappendiceal abscess. Definitive diagnosis and treatment of an infected urachal cyst were made intraoperatively. A review and discussion of urachal remnants is presented, and a diagnostic algorithm and treatment plan is offered for this entity. PMID- 14758309 TI - Edema, anemia, hypoproteinemia, and acrodermatitis enteropathica: an uncommon initial presentation of cystic fibrosis. AB - Cystic fibrosis is a genetic disorder characterized by chronic obstructive pulmonary disease, pancreatic exocrine deficiency, and abnormally high sweat electrolyte concentrations. Less frequently, the presenting features in infants may include edema, anemia, hypoproteinemia, and acrodermatitis enteropathica. Liver involvement may produce hepatomegaly and mild elevation of transaminases. This clinical symptom usually presents within the first 6 months of life and is associated with a high morbidity and mortality. Early recognition and institution of appropriate nutritional supplementation and pancreatic enzymes is essential to improve outcome. Since the sweat test may be falsely negative, emergency physicians must maintain a high index of suspicion to make the diagnosis of cystic fibrosis in an infant who presents with edema, anemia, hypoproteinemia, and acrodermatitis enteropathica. PMID- 14758310 TI - Intravesical catheter knotting: an uncommon complication of urinary catheterization. PMID- 14758311 TI - Retinal hemorrhages in an 8-year-old child: an uncommon presentation of abusive injury. AB - OBJECTIVE: Retinal hemorrhages in pediatric patients have been best described as a component of shaken baby syndrome (SBS), which has been described almost exclusively in the infant/toddler population. We describe the occurrence of retinal hemorrhages in the setting of abusive injury in an older child. METHODS: Case report. RESULTS: An 8-year-old boy was transferred to our institution with coma and respiratory arrest. Evaluation demonstrated intracranial hemorrhage, cerebral edema, and severe bilateral retinal hemorrhages. The patient subsequently died of intractable intracranial hypertension. Police investigation confirmed that the injuries were caused by severe abusive injury, including shaking. CONCLUSIONS: This case emphasizes that the diagnosis of SBS is not limited to babies and that the possibility of abusive shaking injury should also be considered in older children presenting with intracranial pathology and retinal hemorrhages. PMID- 14758312 TI - Coarctation of the aorta and cerebellar infarction: a case report. AB - A 5-year-old child presented to our emergency department with a sudden onset of symptoms consistent with cerebellar infarction. Evaluation lead us to discover an undiagnosed coarctation of the aorta in association with a thrombotic cerebellar infarction. A review of coarctation follows. PMID- 14758313 TI - Variable presentations of rickets in children in the emergency department. AB - Vitamin D-deficient rickets is uncommon but becoming more prevalent in the pediatric population likely related to increases in breast-feeding. It should be considered in many clinical situations. We present 3 cases of rickets presenting acutely to the emergency department. Their presentations included a fracture concerning for child abuse, tetany, and hypocalcemic seizures. In all cases, laboratory and radiographic evaluations were consistent with the diagnosis of nutritional rickets and their symptoms were related to rickets resolved with appropriate treatment. Although uncommon, vitamin D-deficient rickets should be considered in children with the above presentations. PMID- 14758314 TI - Playing basketball, pushed, hip pain, cannot walk. PMID- 14758315 TI - Prehospital emergencies. PMID- 14758316 TI - ECGs in the ED. PMID- 14758317 TI - New oral antihistamines in pediatrics. PMID- 14758318 TI - Calculating the appropriate dose of liquid acetaminophen and ibuprofen. PMID- 14758326 TI - Hidden danger: aortic aneurysms & dissections. PMID- 14758327 TI - Patient-education guide. Heel pain. PMID- 14758328 TI - Helping Iraqi nurses get back on track. Interview by Miriam McCauley. PMID- 14758329 TI - Iron out the details of therapeutic phlebotomy. PMID- 14758330 TI - Hospital-acquired pneumonia: the "gift" that keeps on taking. PMID- 14758331 TI - Confronting posttraumatic stress disorder. PMID- 14758332 TI - The home visit. PMID- 14758334 TI - Abruptio placentae. PMID- 14758333 TI - New drugs 04, part 1. PMID- 14758335 TI - Mutation screening of the phosducin gene PDC in patients with retinitis pigmentosa and allied diseases. AB - PURPOSE: To search for a phenotype associated with mutations in the phosducin gene PDC. METHODS: We screened 853 patients with retinitis pigmentosa or an allied disease diseases, including groups of 61 to 212 patients, each with dominant retinitis pigmentosa (RP), recessive RP, Leber congenital amaurosis, or cone-rod degeneration, for mutations in the PDC gene using direct genomic sequencing of the three coding exons and their flanking intron splice sites. RESULTS: We found one polymorphism in the 5' untranslated region (minor allele frequency of 0.149) and three rare single-base sequence variants (one missense change, one isocoding change, and one in the 3' untranslated region). The rare variants were found in one heterozygous patient each and none was interpreted as pathogenic. CONCLUSIONS: Phosducin mutations are not a major cause of dominant or recessive RP, Leber congenital amaurosis, or cone-rod degeneration. The human phenotype associated with phosducin defects remains unknown. PMID- 14758336 TI - Linkage analysis for age-related macular degeneration supports a gene on chromosome 10q26. AB - PURPOSE: Age-related macular degeneration (AMD) is a retinal degenerative disease that is the leading cause of blindness worldwide in individuals over the age of 60. Although the etiology of AMD remains largely unknown, numerous studies have suggested both genetic and environmental influences. A previous study of affected multiplex families identified four chromosomal regions that potentially harbor AMD susceptibility genes. The purpose of our study was to further investigate these regions with additional microsatellite marker coverage in our independent data set. METHODS: We examined regions on chromosomes 1q, 9p, 10q, and 17q for genetic linkage in our 70 multiplex families (consisting of 133 affected sibpairs). Two point heterogeneity LOD score (HLOD) and nonparametric LOD score (MLS) analyses were performed for disease models defined by the most severe status in either eye. Conditional analyses were performed using apolipoprotein E (APOE) alleles as covariates in semiparametric LOD (LOD*) score calculations. RESULTS: Regions on chromosomes 1q, 9p, and 17q did not provide evidence of linkage in our data set. However, markers D10S1230 and D10S1656 on chromosome 10q26 generated maximum HLOD scores of 1.52 and 1.13, respectively. Marker D10S1230 also generated an MLS score of 1.56 in stage 4 and 5 individuals. Controlling for the potential effect of the APOE-epsilon4 allele did not substantially alter these scores. CONCLUSIONS: With the inclusion of this study, at least five AMD data sets provide support of genetic linkage to 10q26. Such consistency and confirmation of evidence strongly suggests that this region should be the subject of further detailed genomic efforts for the disease. PMID- 14758337 TI - Aquaporin-1 expression is decreased in human and mouse corneal endothelial dysfunction. AB - PURPOSE: To determine if aquaporin-1 expression is decreased in human and mouse corneal endothelial dysfunction. METHODS: Immunohistochemistry with anti aquaporin 1 antibody and confocal microscopy were used to study a case series of human corneal specimens, and a mouse model of corneal endothelial injury was created with injection of sterile hot water into the anterior chamber with a 33 gauge needle. Western blotting of mouse and human corneas was used to provide confirmatory evidence. RESULTS: Aquaporin-1 was found to be expressed in normal human cornea and decreased in human corneas with endothelial disease but not in human corneas with non-endothelial corneal disease. Aquaporin-1 was also found to be reduced in mouse corneas subjected to corneal endothelial injury. Both results were confirmed by western blot analysis. CONCLUSIONS: Corneal endothelial injury is associated with decreased aquaporin-1 expression. PMID- 14758338 TI - The role of VEGF and IGF-1 in a hypercarbic oxygen-induced retinopathy rat model of ROP. AB - PURPOSE: We have previously described a severe form of oxygen-induced retinopathy (OIR) in the neonatal rat, analogous to human retinopathy of prematurity (ROP), where carbon dioxide is added to the inspired environment (hypercarbic OIR). We studied the time course of emergence and resolution of neovascularization (NV) in normocarbic OIR and hypercarbic OIR and the associated changes in VEGF and IGF-1 mRNA levels in hypercarbic OIR. METHODS: 550 newborn Sprague-Dawley rats were raised in 22 expanded litters of 25. Beginning at day 1 of life, rats were exposed to 7 daily cycles of hyperoxia (80% O2, 20.5 h) and hypoxia (10% O2, 0.5 h) with a gradual return to 80% O2 over 3 h. Inspired CO2 was maintained at 0.2% for 200 rats (normocarbic OIR) and 10% for 100 rats (hypercarbic OIR). Rats were sacrificed after a subsequent 5 day room air recovery period. An additional 250 rats raised in room air served as age matched controls. Retinae from left eyes were dissected and flatmounts were ADPase-stained. The presence and severity of NV was scored in a masked manner. Right eyes in hypercarbic OIR litters and room air controls were processed for analysis of VEGF and IGF-1 mRNA. RESULTS: In normocarbic OIR, NV started to emerge before room air recovery began at day 8. It was maximal at day 10 and resolved by day 20. In hypercarbic OIR, a similar pattern was seen, with emergence prior to day 8, peak at day 13 and resolution by day 20. In hypercarbic OIR, retinal VEGF mRNA was decreased at day 8 and increased at day 10 compared to room air controls, correlating with maximal NV. Retinal IGF-1 mRNA was not increased at any time in hypercarbic OIR compared to room air controls. CONCLUSIONS: Neovascularization resulting from normocarbic OIR or hypercarbic OIR occurs before room air recovery. Retinal VEGF mRNA was downregulated and subsequently upregulated prior to maximal NV in hypercarbic OIR. Neovascularization in the hypercarbic OIR model does not appear to be associated with increased retinal IGF-1 mRNA. PMID- 14758339 TI - L-DNase II associated with active process during ethanol induced cell death in ARPE-19. AB - PURPOSE: To analyse the mechanism of ethanol-induced cell death, and particularly, the activation of the leucocyte elastase inhibitor (LEI) pathway. METHODS: Cultured ARPE-19 cells were exposed to 0-13% ethanol for 24 h. Cytotoxicity was estimated by morphologic changes within the nucleus and breakdown of DNA, assessed by agarose gel electrophoresis or flow cytometry cell sorter. Poly(ADP-ribose)polymerase cleavage (PARP) was determined by western blot analysis. Changes in transcription and translation of LEI were assessed by analysis of mRNA levels and expression of protein product (immunohistochemistry), respectively. RESULTS: We established the ability of ethanol to induce cell death in ARPE-19 cells. After a 24 h incubation with 4% ethanol, 50% of the cells died; all the cells died in the presence of 10% ethanol. After ethanol incubation, we observed nuclear condensation and DNA fragmentation; the amount of fragmentation was proportional to the ethanol level. By flow cytometry analysis and agarose gel electrophoresis, the pattern of DNA cleavage exhibited a sub-G1 peak, suggesting necrotic cell death. However, other observations, i.e. nuclei shrinkage, PARP cleavage and inhibition of cell death by cycloheximide, and activation of a caspase independent LEI/DNase II pathway were observed and are features associated with apoptotic cell death. During ethanol stress, an LEI/L-DNase II intermediate was lost, leading to complete activation L-DNase II (24 kDa). RT-PCR analysis showed an early and specific increase of the LEI mRNA. Cycloheximide inhibited LEI synthesis and protected cells against apoptosis. CONCLUSIONS: Our data indicate that ethanol stress on ARPE-19 cells can induce a pathway which is a form of programmed cell death with characteristics of both apoptosis and necrosis, possibly by triggering conversion of LEI to L-DNase II. PMID- 14758340 TI - Immunologic influences on allergy and the TH1/TH2 balance. AB - TH2 cell-mediated immune responses against "innocuous" antigens play a triggering role in atopic allergy. Several epidemiologic studies have clearly shown that the reduced microbial exposure of children caused by the westernized lifestyle is responsible for the increased prevalence of allergy that has occurred in the last decades in developed countries ("hygiene hypothesis"). The immunologic changes caused by the reduced exposure to pathogenic and nonpathogenic microbes during childhood are still controversial. The initial interpretation has been a lack of shift of allergen-specific responses from the TH2 to the TH1 phenotype. This is because of reduced production of IL-12 and IFNs by cells of the natural immunity stimulated by bacterial products through their Toll-like receptors (missing immune deviation). Another interpretation emphasizes the importance of reduced activity of T-regulatory cells (reduced immune suppression). However, although there are impressive amounts of data in favor of the missing immune deviation, experimental evidence supporting the role of reduced immune suppression in explaining the increased prevalence of allergy is currently weak or even contradictory. The solution to this question is very important not only from a theoretic point of view but also because of its therapeutic implications. PMID- 14758341 TI - A potent and selective NPY Y5 antagonist reduces food intake but not through blockade of the NPY Y5 receptor. AB - AIM: These studies were performed to test the hypothesis that endogenous neuropeptide Y (NPY) acting on the NPY Y(5) receptor subtype contributes to the control of food intake. The hypothesis was tested using S 25585-a newly synthesized NPY Y(5) receptor antagonist. METHODS AND RESULTS: S 25585 was shown to be a high-affinity antagonist of the NPY Y(5) receptor subtype (IC(50) 5 nM) with no significant affinity toward other NPY receptor subtypes and over 40 other receptors, channels or uptake systems. S 25585 (7.5 mg/kg, i.p.) did not induce a conditioned taste aversion, significantly alter need-induced sodium appetite or induce pica, suggesting that at this dose the compound did not induce illness or malaise. In satiated rats, S 25585 (5.0 and 7.5 mg/kg, i.p.) significantly decreased the overfeeding induced by i.c.v. injection of NPY (1 microg) and the highly selective NPY Y(5) receptor agonist [hPP(1-17), Ala(31), Aib(32)]NPY (0.7 microg). In rats fasted for 4 h immediately before the dark phase, analysis of the microstructure of feeding behavior revealed that S 25585 significantly increased latency to eat and significantly decreased the duration and size of the meals without altering the meal number or eating rate. Analysis of the behavioral satiety sequence at this time revealed that the animals passed through the normal pattern of feeding, grooming and resting. Although S 25585 appeared to be influencing a physiological system controlling appetite, this does not involve the NPY Y(5) receptor since the antagonist also markedly reduced food intake in the NPY Y(5) knockout mouse. CONCLUSIONS: The results presented do not support a role for the NPY Y(5) receptor in the control of food intake. The results further illustrate that it is imperative that the activity of any new NPY Y(5) antagonist be assessed in the NPY Y(5) knockout mouse before assuming that its effect on food intake is due to blockade of this receptor. PMID- 14758342 TI - Predictors of body size in the first 2 y of life: a high-risk study of human obesity. AB - OBJECTIVE: To ascertain the predictors of body size at 2 y of age. DESIGN: : Prospective, longitudinal study of risk factors for weight gain of infants at high or low risk of obesity by virtue of their mothers' obesity or leanness. SUBJECTS: A total of 40 infants of obese mothers and 38 infants of lean mothers, equally divided among boys and girls. METHODS: Measurement of dependent variables: weight, length and skinfold thicknesses at 3, 6, 9, 12, 18 and 24 months and percent body fat at 3, 12 and 24 months. Measurement of independent variables: average daily caloric consumption at 3, 6, 9, 12, 18 and 24 months; and, at 3 months, nutritive sucking behavior during a test meal, total energy expenditure (TEE), sleeping energy expenditure (SEE), estimation of nonsleeping energy expenditure (TEE-SEE) and socioeconomic status. Parental weights and heights were obtained by self-report at the time of recruitment. Partial correlation and mixed effects linear regression analyses were performed. RESULTS: Measures of body size (weight, length, skinfold thicknesses) and percent of body fat were almost identical between high- and low-risk groups at all times. Energy intake during six occasions over the 2 y, sucking behavior, family income and TEE predicted weight gain, controlling for body length. Parental body mass index was not associated with the child's body size during the first 2 y. During the first year, there were strong lagged correlations between energy intake and body weight and smaller correlations between protein intake and body weight. CONCLUSION: Energy intake, and not energy expenditure, was the determinant of body size in these infants at 2 y of age, as it had been at 1 y. Sucking behavior and TEE (positively) and family income (negatively) also contributed to body weight at 2 y. The novel finding of a lagged correlation between energy intake and body weight early in life suggests that energy intake is programmed for future growth and development. PMID- 14758343 TI - Expression of estrogen receptors in the coronary arteries of young and premenopausal women in relation to central obesity. AB - OBJECTIVE: To examine the relationship of coronary estrogen receptor (ER) expression with atherosclerotic lesions and central fat accumulation in premenopausal women. SUBJECTS: A total of 52 female forensic autopsy cases aged between 18 and 49 y. METHODS: Height, body weight and waist and hip circumferences were measured and body mass index (BMI) and waist-to-hip ratio (WHR) were calculated. Intima thickness or maximal thickness of the plaque were measured from samples taken from the left anterior descending artery (LAD). Macrophage infiltration and smooth muscle cells were localized by immunostaining. ER was detected immunohistochemically and by Western blot analysis, and the ER immunopositive area in the intima was measured. RESULTS: ER immunoreactivity was observed in the intima in 60% of the samples, and it was most intense in the advanced plaques near the lipid core next to the maximal intensity of macrophage staining. The ER immunopositive area had a significant positive correlation with LAD intima thickness, which in turn was significantly correlated with waist circumference and WHR when adjusted for age and BMI. CONCLUSIONS: Premenopausal women with the central type of fat accumulation have advanced coronary plaques in which ER expression is localized near the lipid-rich and macrophage-rich zone. The higher expression of ER in the arterial plaques may represent a compensatory mechanism against atherosclerosis. PMID- 14758344 TI - Activation of leptin expression by an inhibitor of carnitine palmitoyltransferase 1. AB - Leptin may regulate peripheral fatty acid oxidation and invoke a feedback mechanism that affects leptin expression in adipocytes. The objective of this study, therefore, was to determine whether inhibiting systemic fatty acid oxidation at the level of carnitine palmitoyltransferase-1 (CPT1) affects leptin expression. To accomplish this objective, fed or overnight fasted rats were treated with 2-tetradecylglycidic acid (TDGA), a specific, irreversible CPT1 inhibitor, and acute changes in rat epididymal leptin expression and serum leptin content were measured using Northern, RT-PCR, and radioimmunoassay analyses. Overnight fasting decreased both epididymal leptin mRNA content and serum leptin. Treating overnight fasted rats with TDGA increased both their epididymal leptin mRNA and their serum leptin significantly in a time- and concentration-dependent manner. TDGA affected neither epididymal leptin mRNA nor serum leptin in fed rats where systemic fatty acid oxidation is low. These results support the conclusion that CPT1-linked fatty acid oxidation is a key modulator of leptin expression in fasting rats. PMID- 14758345 TI - Association between stunting and overweight among 10-15-y-old children in the North West Province of South Africa: the THUSA BANA Study. AB - OBJECTIVE: To investigate the relationship between stunting and overweight among 10-15-y-old children of the North West Province in South Africa. DESIGN: A single cross-sectional study design was used. The study formed part of the THUSA BANA project. SUBJECTS: The total study population of the THUSA BANA project comprised of 1257 randomly selected subjects, aged 10-15 y. MEASUREMENTS: Stunting was described as the height below the 5th percentile for age using the CDC standard percentiles. Furthermore, the definitions of overweight and obesity according to the International Obesity Task Force (IOTF) were used, where the cutoff points for body mass index (BMI) corresponds with the adult BMI of 25 and 30, respectively. Anthropometrical variables namely triceps (TSF) and subscapular skinfolds (SSF), waist circumference, weight, height and BMI of the 10-15-y-old subjects were analysed. RESULTS: Stunting was most prevalent in the rural areas (girls 23.7% and boys 26.7%) compared with urban areas (girls 11.6%, boys 17.1%). The odds ratio and the 95% CI for the association between stunting and overweight in boys and girls were 0.45 (CI 0.16, 1.30) and 0.50 (CI 0.21, 1.19) respectively. Stunted children, 10-14-y-old and living in rural areas and informal settlements, had significantly lower mean BMI and skinfold thicknesses than nonstunted children. The mean BMI and sum of TSF and SSF (TSF+SSF) were similar in stunted and nonstunted children living in urban areas. CONCLUSION: There is no significant association between stunting and overweight in 10-15-y old children in the North West Province. However, there is a tendency for girls older than 14 y to start to gain subcutaneous fat, even though at these ages they were still stunted and underweight. Stunted girls in established urban areas had a higher mean TSF+SSF than stunted girls in informal townships. This tendency in urban stunted girls is evident at the onset of menarche and could predict possible problems of overweight as they get older. PMID- 14758346 TI - Similarity of cutaneous reactive hyperemia in the forearm of women with and without hyperinsulinemia. AB - OBJECTIVE: The present study was undertaken to assess in pre- and postmenopausal women whether obesity influences cutaneous reactive hyperemia in the forearm. DESIGN: Cross-sectional study. SUBJECTS AND METHODS: Eight lean premenopausal (age 24.6+/-3.5 y, BMI=21.9+/-1.5 kg/m(2), mean+/-1 s.d.), eight obese premenopausal (age 27.8+/-5.1 y, BMI=35.3+/-5.8 kg/m(2)), eight lean postmenopausal (age 56.1+/-8.3 y, BMI=20.2+/-2.2 kg/m(2)) and eight obese postmenopausal women (age 57.4+/-6.1 y, BMI=32.8+/-3 kg/m(2)) were included. Plasma glucose, insulin and lipid profile were determined in fasting state, and a glucose tolerance test was performed. The skin blood flow response to transient occlusion of the forearm circulation (reactive hyperemia, RH) was measured using a laser-Doppler imaging system. RESULTS: Obese women had hyperinsulinemia, suggesting that they were insulin-resistant. The magnitude of the RH was similar in postmenopausal compared to premenopausal women. Obesity did not influence this microvascular response. CONCLUSION: Obesity, which is known to be associated with impaired endothelial-dependent vasodilation in the skeletal musculature, has no effect on the RH of the forearm skin microcirculation. PMID- 14758347 TI - Uncoupling proteins: gender dependence and their relation to body weight control. AB - Members of the uncoupling protein (UCP) family have different purported functions, which can be either directly or indirectly related to the control of body weight. In this sense, most studies on this topic have been carried out using male subjects, although different works with males and females have shown important sex-associated differences in the regulation of these proteins; for instance, sex differences have been shown in the cold-, diet- and overweight induced expression of brown adipose tissue UCP1 and also in the correlation of muscle UCP3 with overweight. In these kinds of studies, models of obesity such as cafeteria diet feeding and postcafeteria have been very useful. Moreover, sex hormones have been shown to modulate UCP1 expression in brown adipocytes in vitro. All these sex-dependent differences, as well as sex differences in body weight gain under a hypercaloric diet, could be related to the different respective biological functions of males and females, taking into account the fact that the gender effect in future studies on obesity could be of interest. PMID- 14758348 TI - Reduced sodium output following acute spinal injury. AB - STUDY DESIGN: Assessment of sodium output in spinal injury patients. OBJECTIVES: The purpose was to examine the effects of sodium loading acutely by an infusion, long term by sodium supplements, to acutely injured spinal patients on a fixed sodium intake. This was compared with another group of acutely injured patients who were on a hospital diet of between 50 and 150 mmol of sodium daily. SETTING: The National Spinal Injuries Centre, Stoke Mandeville Hospital, Aylesbury, Bucks HP21 8AL, UK. METHODS: A total of 53 studies were carried out on 52 patients between 1962 and 1964. In all, 49 patients were studied during the first 10 days after injury as follows: a control group of 39 patients received no supplementary sodium, two received NaCl infusion, two NaCl tablets orally, and six received NaCl infusion followed by NaCl tablets orally. Four patients were studied for more than 15 days after injury (one of these had been studied in the acute stage); they received NaCl infusion and one (42d) received in addition NaCl tablets for four subsequent days. RESULTS: In all patients urinary sodium excretion was minimal on day 2 and increased thereafter. On days 2-6, it was significantly lower in patients with a complete transection of the cervical cord than in patients with lower lesions. In the early studies, nine patients excreted less than 40% of the administered load within 24 h. In four of these patients excretion was 10% or less. In the later studies, three of the four patients excreted at least 80% of the infused Na+ on the same day. CONCLUSION: Sodium retention in the patient with cord injury is a response to trauma. The different responses seen in patients with different levels of cord transection are not due to direct changes in the innervation of the kidney but to compensation for sympathetic insufficiency, blood pressure being maintained by the secretion of aldosterone, vasopressin and other hormones. The decreased urinary output seen acutely after cord transection is not due to renal failure and the patient's condition can be made dangerously worse by attempts to create a diuresis. PMID- 14758349 TI - Perceived outcomes and utilization of upper extremity surgical reconstruction in individuals with tetraplegia at model spinal cord injury systems. AB - STUDY DESIGN: Survey. OBJECTIVE: To measure the utilization of upper extremity reconstructive surgery and the clinicians' perceptions of the outcomes provided for persons with tetraplegia across the Model Spinal Cord (SCI) Injury Systems. SETTING: Model SCI Systems. METHODS: PARTICIPANTS: A clinician from each of the Model Centers. MAIN OUTCOME MEASURE: A mailed survey eliciting responses with respect to: (1) utilization of upper extremity reconstructive procedures and (2) the clinicians' perceived outcomes of these procedures. RESULTS: In all, 76% responded positively about the availability and appropriateness of upper extremity surgical reconstruction at their center. Of the respondents, 75% felt that surgery recipients were generally satisfied with their surgeries, 80% felt that the surgery made a positive impact on recipients' lives, 81% felt that recipients showed increased independence, and 70% reported a positive impact on recipients' occupation. In all, 93% felt insurance companies should pay for the procedures. Compared to the satisfaction of surgery recipients using a similar instrument, clinicians anticipated slightly greater improvements in all areas except occupation. CONCLUSIONS: There is a positive perception of the benefits of reconstructive surgery for tetraplegia; however, procedures are not routinely offered at all centers. The primary reasons reported for this include the misconception that insurance does not remit payment, that a surgeon is not available, and that surgical candidates are referred to another center. PMID- 14758350 TI - Role of NMDA receptor activation in serotonin agonist-induced air-stepping in paraplegic mice. AB - STUDY DESIGN: Experimental laboratory investigation of the effects of serotonergic and glutamatergic drugs in early paraplegic mice. OBJECTIVES: To examine whether NMDA and 5-HT receptors synergistically participate to generate basic stepping movements in paraplegic mice. SETTING: Laval University Medical Center, Quebec, Canada. METHODS: Adult mice completely spinalized at the low thoracic level 1 week earlier were suspended in harnesses for experiments. Acute drug-induced effects were examined on hindlimb movements filmed with a digital video camera. Detailed kinematic analyses included stick diagrams reconstructions of hindlimb movements and analysis of bilateral coordination, angular excursion, stepping amplitude and frequency. RESULTS: A single treatment with the 5-HT2 agonist quipazine (>0.7 mg/kg, i.p.) induced episodes of air-stepping movements in the hindlimbs of paraplegic mice. In contrast, injection of the glutamatergic agonist NMDA (1-45 mg/kg i.p.) failed to induce rhythmicity, although nonlocomotor rhythmic movements were observed with higher doses (45-60 mg/kg i.p.). Subthreshold doses of NMDA (22-30 mg/kg) could induce episodes of hindlimb air-stepping if combined with subthreshold doses of quipazine (0.3-0.7 mg/kg). Air-stepping was entirely blocked by administration of the selective NMDA antagonist MK-801. CONCLUSION: A single treatment with quipazine can trigger episodes of locomotor-like movements in early chronic spinal mice. Even though NMDA alone could not generate bilaterally coordinated air-stepping, NMDA receptor activation was nonetheless critical for spinal locomotor rhythmogenesis induced by 5-HT agonists in awake behaving animals. PMID- 14758351 TI - Vocational reintegration following spinal cord injury: expectations, participation and interventions. AB - STUDY DESIGN: Survey. OBJECTIVES: To explore the process of reintegration in paid work following a traumatic spinal cord injury (SCI), including the role of early expectations of individual patients regarding return to work, indicators of success of job reintegration and a description of reintegration interventions and barriers. SETTING: Dutch rehabilitation centre with special department for patients with spinal cord injuries. METHODS: Descriptive analysis of data gathered by a mailed questionnaire, which was returned by 57 persons (response 83%) with traumatic SCI, aged 18-60 years, and data of earlier expectations reported by the individual patients during the rehabilitation admission following SCI from 1990 to 1998. RESULTS: Of 49 respondents who were employed at the moment of the SCI, 45% expected to be able to resume work. These positive expectations were associated with a higher educational level. In 67%, return to work was successful. The chance to reintegrate successfully was better if the patient expected to resume work. Logistic regression analysis did not reveal other significant indicators. About one-third of the 49 respondents working preinjury followed vocational retraining, which was successful for most of them so far. In the majority of work situations modifications have been made, such as job adaptations and reduction of working hours. Several unmet needs regarding reintegration interventions were also reported. CONCLUSIONS: Positive expectations regarding resumption of work after a SCI are an important indicator of successful reintegration in work. An active role of the rehabilitation team is recommended in drawing up a vocational reintegration plan to prepare the patient, the employer and professionals involved in the reintegration process. PMID- 14758352 TI - Intravesical therapy options for neurogenic detrusor overactivity. AB - STUDY DESIGN: Review article. SETTING: Neuro-Urology, Spinal Cord Injury Center, Balgrist University Hospital, Zurich, Switzerland. OBJECTIVES: This review considers intravesical treatment options of neurogenic detrusor overactivity and discusses the underlying mechanism of action, clinical safety and efficacy, and the future trends. METHODS: The available literature was reviewed using medline services. RESULTS: Oral anticholinergic drugs are widely used to treat detrusor overactivity, but they are ineffective in some patients or cause systemic side effects such as blurred vision or dry mouth. As an alternative, topical therapy strategies have been suggested to achieve a profound inhibition of the overactive detrusor and to avoid high systemic drug levels. Currently available intravesical treatment options either act on the afferent arc of the reflex such as local anaesthetics or vanilloids or on the efferent cholinergic transmission to the detrusor muscle such as intravesical oxybutynin or botulinum toxin. Although an established and effective therapy, intravesical oxybutynin is not widely used. Evidence for clinical significance of intravesical atropine and local anaesthetic is missing. Intravesical capsaicin has been shown to improve clinical and urodynamic parameters, but cause pain in some patients. The intravesical instillation of resiniferatoxin and the injection of botulinum-A toxin into the detrusor muscle are promising new options; however, randomised placebo-controlled studies to prove their safety and efficacy are still missing. CONCLUSION: Intravesical treatment strategies in patients with neurogenic detrusor overactivity may provide alternatives to established therapies such as oral anticholinergics. The selectivity of the intravesical treatment and the reduction or even the absence of side effects are major advantages of this topical approach. PMID- 14758353 TI - A DNA enzyme that mimics the first step of RNA splicing. AB - We have discovered an artificial DNA enzyme that mimics the first step of RNA splicing. In vitro selection was used to identify DNA enzymes that ligate RNA. One of the new DNA enzymes carries out splicing-related catalysis by specifically recognizing an unpaired internal adenosine and facilitating attack of its 2' hydroxyl onto a 5'-triphosphate. This reaction forms 2',5'-branched RNA and is analogous to the first step of in vivo RNA splicing, in which a ribozyme cleaves itself with formation of a branched intermediate. Unlike a natural ribozyme, the new DNA enzyme has no 2'-hydroxyl groups to aid in the catalytic mechanism. Our finding has two important implications. First, branch-site adenosine reactivity seems to be mechanistically favored by nucleic acid enzymes. Second, hydroxyl groups are not obligatory components of nucleic acid enzymes that carry out biologically related catalysis. PMID- 14758354 TI - Crystal structure of Dcp1p and its functional implications in mRNA decapping. AB - A major pathway of eukaryotic mRNA turnover begins with deadenylation, followed by decapping and 5'-->3' exonucleolytic degradation. A critical step in this pathway is decapping, which is carried out by an enzyme composed of Dcp1p and Dcp2p. The crystal structure of Dcp1p shows that it markedly resembles the EVH1 family of protein domains. Comparison of the proline-rich sequence (PRS)-binding sites in this family of proteins with Dcp1p indicates that it belongs to a novel class of EVH1 domains. Mapping of the sequence conservation on the molecular surface of Dcp1p reveals two prominent sites. One of these is required for the function of the Dcp1p-Dcp2p complex, and the other, corresponding to the PRS binding site of EVH1 domains, is probably a binding site for decapping regulatory proteins. Moreover, a conserved hydrophobic patch is shown to be critical for decapping. PMID- 14758355 TI - Role of teichoic acids in Staphylococcus aureus nasal colonization, a major risk factor in nosocomial infections. AB - Colonization of the anterior nares in approximately 37% of the population is a major risk factor for severe Staphylococcus aureus infections. Here we show that wall teichoic acid (WTA), a surface-exposed staphylococcal polymer, is essential for nasal colonization and mediates interaction with human nasal epithelial cells. WTA-deficient mutants were impaired in their adherence to nasal cells, and were completely unable to colonize cotton rat nares. This study describes the first essential factor for S. aureus nasal colonization. PMID- 14758356 TI - Activation of nuclear hormone receptor peroxisome proliferator-activated receptor delta accelerates intestinal adenoma growth. AB - We treated Apc(min) mice, which are predisposed to intestinal polyposis, with a selective synthetic agonist of peroxisome proliferator-activated receptor-delta (PPAR-delta). Exposure of Apc(min) mice to the PPAR-delta ligand GW501516 resulted in a significant increase in the number and size of intestinal polyps. The most prominent effect was on polyp size; mice treated with the PPAR-delta activator had a fivefold increase in the number of polyps larger than 2 mm. Our results implicate PPAR-delta in the regulation of intestinal adenoma growth. PMID- 14758357 TI - B cell receptor signal strength determines B cell fate. AB - B cell receptor (BCR)-mediated antigen recognition is thought to regulate B cell differentiation. BCR signal strength may also influence B cell fate decisions. Here, we used the Epstein-Barr virus protein LMP2A as a constitutively active BCR surrogate to study the contribution of BCR signal strength in B cell differentiation. Mice carrying a targeted replacement of Igh by LMP2A leading to high or low expression of the LMP2A protein developed B-1 or follicular and marginal zone B cells, respectively. These data indicate that BCR signal strength, rather than antigen specificity, determines mature B cell fate. Furthermore, spontaneous germinal centers developed in gut-associated lymphoid tissue of LMP2A mice, indicating that microbial antigens can promote germinal centers independently of BCR-mediated antigen recognition. PMID- 14758358 TI - Regulatory T cells mediate maternal tolerance to the fetus. AB - Pregnancy constitutes a major challenge to the maternal immune system, as it has to tolerate the persistence of paternal alloantigen. Although localized mechanisms contribute to fetal evasion from immune attack, maternal alloreactive lymphocytes persist. We demonstrate here an alloantigen-independent, systemic expansion of the maternal CD25+ T cell pool during pregnancy and show that this population contains dominant regulatory T cell activity. In addition to their function in suppressing autoimmune responses, maternal regulatory T cells suppressed an aggressive allogeneic response directed against the fetus. Their absence led to a failure of gestation due to immunological rejection of the fetus. PMID- 14758359 TI - ERM proteins regulate cytoskeleton relaxation promoting T cell-APC conjugation. AB - During activation, T cells associate with antigen-presenting cells, a dynamic process that involves the formation of a broad area of intimate membrane contact known as the immunological synapse. The molecular intermediates that link initial antigen recognition to the cytoskeletal changes involved in this phenomenon have not yet been defined. Here we demonstrate that ezrin-radixin-moesin proteins are rapidly inactivated after antigen recognition through a Vav1-Rac1 pathway. The resulting disanchoring of the cortical actin cytoskeleton from the plasma membrane decreased cellular rigidity, leading to more efficient T cell-antigen presenting cell conjugate formation. These findings identify an antigen-dependent molecular pathway that favors immunological synapse formation and the subsequent development of an effective immune response. PMID- 14758360 TI - Stochastic yet biased expression of multiple Dscam splice variants by individual cells. AB - The Drosophila melanogaster gene Dscam is essential for axon guidance and has 38,016 possible alternative splice forms. This diversity can potentially be used to distinguish cells. We analyzed the Dscam mRNA isoforms expressed by different cell types and individual cells. The choice of splice variants expressed is regulated both spatially and temporally. Different subtypes of photoreceptors express broad yet distinctive spectra of Dscam isoforms. Single-cell RT-PCR documented that individual cells express several different Dscam isoforms and allowed an estimation of the diversity that is present. For example, we estimate that each R3/R4 photoreceptor cell expresses 14-50 distinct mRNAs chosen from the spectrum of thousands of splice variants distinctive of its cell type. Thus, the Dscam repertoire of each cell is different from those of its neighbors, providing a potential mechanism for generating unique cell identity in the nervous system and elsewhere. PMID- 14758361 TI - Mutant P450 oxidoreductase causes disordered steroidogenesis with and without Antley-Bixler syndrome. AB - Deficient activities of multiple steroidogenic enzymes have been reported without and with Antley-Bixler syndrome (ABS), but mutations of corresponding cytochrome P450 enzymes have not been found. We identified mutations in POR, encoding P450 oxidoreductase, the obligate electron donor for these enzymes, in a woman with amenorrhea and three children with ABS, even though knock-out of POR is embryonically lethal in mice. Mutations of POR also affect drug-metabolizing P450 enzymes, explaining the association of ABS with maternal fluconazole ingestion. PMID- 14758362 TI - Evolutionary innovation of the excretory system in Caenorhabditis elegans. AB - The evolution of complexity relies on changes that result in new gene functions. Here we show that the unique morphological and functional features of the excretory duct cell in C. elegans result from the gain of expression of a single gene. Our results show that innovation can be achieved by altered expression of a transcription factor without coevolution of all target genes. PMID- 14758363 TI - The hyh mutation uncovers roles for alpha Snap in apical protein localization and control of neural cell fate. AB - The hyh (hydrocephalus with hop gait) mouse shows a markedly small cerebral cortex at birth and dies postnatally from progressive enlargement of the ventricular system. Here we show that the small hyh cortex reflects altered cell fate. Neural progenitor cells withdraw prematurely from the cell cycle, producing more early-born, deep-layer cerebral cortical neurons but depleting the cortical progenitor pool, such that late-born, upper-layer cortical neurons are underproduced, creating a small cortex. hyh mice carry a hypomorphic missense mutation in the gene Napa encoding soluble N-ethylmaleimide-sensitive factor (NSF) attachment protein alpha (alpha Snap), involved in SNAP receptor (SNARE) mediated vesicle fusion in many cellular contexts. A targeted null Napa mutation is embryonically lethal. Altered neural cell fate is accompanied by abnormal localization of many apical proteins implicated in regulation of neural cell fate, including E-cadherin, beta-catenin, atypical protein kinase C (aPKC) and INADL (inactivation-no-afterpotential D-like, also known as protein associated with Lin7, or Pals1). Apical localization of the SNARE Vamp7 is also disrupted. Thus, alpha Snap is essential for apical protein localization and cell fate determination in neuroepithelial cells. PMID- 14758364 TI - Encoding of emotional memories depends on amygdala and hippocampus and their interactions. AB - We have studied patients with variable degrees of left hippocampal and amygdala pathology who performed a verbal encoding task during functional magnetic resonance imaging (fMRI) to assess the impact of pathology on emotional-memory performance and encoding-evoked activity. The severity of left hippocampal pathology predicted memory performance for neutral and emotional items alike, whereas the severity of amygdala pathology predicted memory performance for emotional items alone. Encoding-related hippocampal activity for successfully remembered emotional items correlated with the degree of left amygdala pathology. Conversely, amygdala-evoked activity with respect to subsequently remembered emotional items correlated with the degree of left hippocampal pathology. Our data indicate a reciprocal dependence between amygdala and hippocampus during the encoding of emotional memories. PMID- 14758365 TI - In vivo imaging of synapse formation on a growing dendritic arbor. AB - The form of a neuron's dendritic arbor determines the set of axons with which it may form synaptic contacts, thus establishing connectivity within neural circuits. However, the dynamic relationship between dendrite growth and synaptogenesis is not well understood. To observe both processes simultaneously, we performed long-term imaging of non-spiny dendritic arbors expressing a fluorescent postsynaptic marker protein as they arborized within the optic tectum of live zebrafish larvae. Our results indicate that almost all synapses form initially on newly extended dendritic filopodia. A fraction of these nascent synapses are maintained, which in turn stabilizes the subset of filopodia on which they form. Stabilized filopodia mature into dendritic branches, and successive iterations of this process result in growth and branching of the arbor. These findings support a 'synaptotropic model' in which synapse formation can direct dendrite arborization. PMID- 14758366 TI - Rational siRNA design for RNA interference. AB - Short-interfering RNAs suppress gene expression through a highly regulated enzyme mediated process called RNA interference (RNAi). RNAi involves multiple RNA protein interactions characterized by four major steps: assembly of siRNA with the RNA-induced silencing complex (RISC), activation of the RISC, target recognition and target cleavage. These interactions may bias strand selection during siRNA-RISC assembly and activation, and contribute to the overall efficiency of RNAi. To identify siRNA-specific features likely to contribute to efficient processing at each step, we performed a systematic analysis of 180 siRNAs targeting the mRNA of two genes. Eight characteristics associated with siRNA functionality were identified: low G/C content, a bias towards low internal stability at the sense strand 3'-terminus, lack of inverted repeats, and sense strand base preferences (positions 3, 10, 13 and 19). Further analyses revealed that application of an algorithm incorporating all eight criteria significantly improves potent siRNA selection. This highlights the utility of rational design for selecting potent siRNAs and facilitating functional gene knockdown studies. PMID- 14758367 TI - In Methuselah's Mould. PMID- 14758370 TI - A major pitfall in the search strategy on PubMed. PMID- 14758368 TI - Contribution of noncentrosomal microtubules to spindle assembly in Drosophila spermatocytes. AB - Previous data suggested that anastral spindles, morphologically similar to those found in oocytes, can assemble in a centrosome-independent manner in cells that contain centrosomes. It is assumed that the microtubules that build these acentrosomal spindles originate over the chromatin. However, the actual processes of centrosome-independent microtubule nucleation, polymerisation, and sorting have not been documented in centrosome-containing cells. We have identified two experimental conditions in which centrosomes are kept close to the plasma membrane, away from the nuclear region, throughout meiosis I in Drosophila spermatocytes. Time-lapse confocal microscopy of these cells labelled with fluorescent chimeras reveals centrosome-independent microtubule nucleation, growth, and sorting into a bipolar spindle array over the nuclear region, away from the asters. The onset of noncentrosomal microtubule nucleation is significantly delayed with respect to nuclear envelope breakdown and coincides with the end of chromosome condensation. It takes place in foci that are close to the membranes that ensheath the nuclear region, not over the condensed chromosomes. Metaphase plates are formed in these spindles, and, in a fraction of them, some degree of polewards chromosome segregation takes place. In these cells that contain both membrane-bound asters and an anastral spindle, the orientation of the cytokinesis furrow correlates with the position of the asters and is independent of the orientation of the spindle. We conclude that the fenestrated nuclear envelope may significantly contribute to the normal process of spindle assembly in Drosophila spermatocytes. We also conclude that the anastral spindles that we have observed are not likely to provide a robust back-up able to ensure successful cell division. We propose that these anastral microtubule arrays could be a constitutive component of wild-type spindles, normally masked by the abundance of centrosome-derived microtubules and revealed when asters are kept away. These observations are consistent with a model in which centrosomal and noncentrosomal microtubules contribute to the assembly and are required for the robustness of the cell division spindle in cells that contain centrosomes. PMID- 14758371 TI - Food poisoning in Saudi Arabia. Potential for prevention? AB - Food poisoning is becoming a very important health problem both internationally and in the Kingdom of Saudi Arabia (KSA). Salmonella species (spp) is the most important pathogen, but due to the extensive effort carried by the health authorities to prevent or eradicate communicable diseases, new pathogens are now emerging such as Escherichia coli (E. coli) and Norwalk like viruses. This review paper highlights the magnitude and determinants of food poisoning internationally and in KSA, and proposes some recommendations on its prevention. Clearly, there is a steady increase in the food poisoning accidents in KSA, especially during the summer months and Hajj season. These accidents are seen in other countries such as England, United States of America and Japan. Meat and chicken are the main items incriminated in these accidents. Knowledge on food safety in the food preparation process and the risk factors that can lead to food poisoning is low. Training and proper health education messages are needed to raise the awareness of food handlers as well as the public in general. PMID- 14758372 TI - Cancer-related anemia. AB - Anemia is the most common hematological abnormality in cancer patients, unfortunately, it is often under-recognized and under-treated. The pathogenesis of cancer anemia is complex and most of the time multifactorial; involving factors related to the tumor itself or its therapy. While anemia can present in a wide range of symptoms, involving almost every organ, it is believed that it contributes much to cancer-related fatigue, one of the most common symptoms in cancer patients. In addition, there is increasing evidence to suggest that anemia is an independent factor adversely affecting tumor response and patient survival. While blood transfusion was the only option to treat cancer-related anemia, the use of recombinant human erythropoietin (rHuEPO) is becoming the new standard of care, more so with the recent studies demonstrating the feasibility of a single weekly injection. Things are even getting better with the recent approval of a new form of rHuEPO; Darbepoetin, an analogue with a 3-fold longer half-life. In addition to its effect in raising hemoglobin, several well-controlled studies demonstrated decrease in transfusion requirements and better quality of life assessed objectively using standard assessments scales. PMID- 14758373 TI - Enterococcal bacteremia in a teaching hospital in the Central region of Saudi Arabia. AB - OBJECTIVE: To study enterococcal blood stream infection including the different species isolated, their antibiotic resistance associated risk factors and outcome of treatment. METHODS: A retrospective and prospective study was carried out over a 12 month period between June 2001 and May 2002 in King Khalid University Hospital, Riyadh, Kingdom of Saudi Arabia. Sixty episodes of enterococcal bacteremia with clinical significance detected in adults and neonates were included. Statistical analysis of the epidemiological characteristic, etiologic risk factors and mortality were determined. RESULTS: Fifty-four (90%) of the episodes were hospital acquired. Fifty-five percent of patients were elderly males and 20% were neonates. Patients usually have severe underlying diseases (57%) and the mortality rate was 28%. The estimated duration of hospital stay after acquiring the Enterococcal bacteremia was 40 days. The source of bacteremia remained undetermined in 72% of the episodes. Gastrointestinal tract, urinary tract and intravascular catheter were the most frequently recognized associated sites of infection. Thirty-three patients (55%) had previously received antimicrobial agents (mainly cephalosporins). In this study, intravascular catheter (p=0.0002), urinary catheter (p=0.00001), mechanical ventilation (p=0.002), previous surgery (digestive tract surgery (p=0.01) and prior stay in an intensive care unit (p=0.03) were the factors associated with Enterococcal bacteremia and mortality. CONCLUSION: Efforts to reduce the occurrence of enterococcal bacteremia should be focused on appropriate use of cephalosporins and external devices. PMID- 14758374 TI - Prevalence of HBV, HCV, HIV-1, 2 and HTLV-I/II infections among blood donors in a teaching hospital in the Central region of Saudi Arabia. AB - OBJECTIVE: Several infectious diseases are transmissible by blood transfusion, especially viral infections. The most common blood-transmitted viruses are hepatitis B virus (HBV), hepatitis C virus (HCV) and human immunodeficiency virus (HIV). These viruses cause fatal, chronic and life-threatening disorders. The prevalence of these viruses varies by nationality and geography. The purpose of this study was to establish the current prevalence of hepatitis viruses (B and C) and human retroviruses (HIV-1, 2 and human T-lymphotropic virus type I and II, HTLV-I /II) among blood donors at King Khalid University Hospital (KKUH), Riyadh, Kingdom of Saudi Arabia (KSA). METHODS: Serological markers of HBV, HCV, HIV 1, 2 and HTLV-I/II were studied in 24173 (23952 males and 221 females), 20423 Saudi and 3750 non-Saudi blood donors, using commercially available kits, over a period of 3 years from January 2000 to December 2002 at KKUH, Riyadh, KSA. The prevalence of confirmed-positive test results of these viruses was evaluated among different gender, ages and nationalities. RESULTS: During the study period, prevalence rates of HBV and HCV infections were 1.5% and 0.4%, and zero for retroviral infections. The prevalence was not significantly higher in male than in female donors. Hepatitis B surface antigen (HBsAg) and anti-HCV positivity tend to increase with increase in age. The prevalence of HBsAg and anti-HCV positivity was significantly more prevalent among non-Saudi compared to Saudi donors. CONCLUSION: This study highlights the prevalence rates of HBV and HCV among different groups. The prevalence varies from one group to another, being the lowest among Saudi and young donors. Therefore, extensive recruitment of Saudi and young donors should help ensure a long-term increase in the blood supply without jeopardizing safety. PMID- 14758375 TI - Intestinal parasites among presumably healthy individuals in Lebanon. AB - OBJECTIVE: To determine the prevalence period of intestinal parasites among presumably healthy subjects in Lebanon. METHODS: One stool specimen from 2634 presumably healthy Lebanese subjects, mean age 32.1 years with a range of 14-71 years, resident of different areas in Lebanon was examined for the presence of parasites. The analysis took place in the Clinical Microbiology Laboratory, Department of Pathology and Laboratory Medicine, American University of Beirut Medical Center, Beirut, Lebanon, over 25-months between 1995-1997. RESULTS: The prevalence of intestinal parasites was 12.4%. The most common parasites identified were Entamoeba coli (38%), Giardia lamblia (3.1%) and Entamoeba histolytica (2.3%) [corrected]. CONCLUSION: The data presented shows the need to improve hygienic conditions to contain the problem of intestinal infections with parasites in Lebanon. PMID- 14758376 TI - Prophylactic ciprofloxacin drops after tympanostomy tube insertion. AB - OBJECTIVE: To evaluate the effectiveness of prophylactic ciprofloxacin drops in decreasing the incidence of post-tympanostomy otorrhea, and the relation between middle ear content and post-tympanostomy otorrhea. METHODS: One hundred and fifty patients aged 3-14 years underwent tympanostomy and tube insertion at the Prince Rashid Ben Al-Hasan Hospital, Al-Husn, Jordan during the interval between February 2000 to January 2003. The patients were randomized into 3 groups: group 1 (control group) received no antibiotic drops, group 2 received a single dose of ciprofloxacin drops intraoperatively and group 3 received an intraoperative dose followed by 5-day postoperative course. RESULTS: Application of topical ciprofloxacin after tympanostomy tube insertion was associated with a significantly lower incidence of early post-tympanostomy otorrhea. The rate of otorrhea for the control group was 16.5% and the treatment groups were (group 1) 8.4%, (group 2) 8.2% and p=0.011. A single dose of antibiotics was effective when patient's middle ears were dry or had serous effusions. For those whose ears had mucoid or purulent content a 5-day course was indicated. CONCLUSION: Topical administration of a single dose of ototopical ciprofloxacin after surgery is an effective treatment for the prevention of early post-tympanostomy otorrhea, and a prolonged course (5 days) may be indicated for those whose ears had purulent or thick mucoid contents, and the content of middle ear are important in predicating postoperative otorrhea. PMID- 14758377 TI - Ultrasonography of the uterus after normal vaginal delivery. AB - OBJECTIVE: To define the appearance of the uterus and the uterine cavity, as revealed by ultrasound in normal women following a vaginal delivery. METHODS: This prospective, longitudinal study took place at the Prince Hashem and Prince Ali Military Hospitals, Amman, Jordan from December 2002 to March 2003. Fifty four women were scanned on postpartum days 1, 7, 14, 28, and 56. Ultrasound examination was performed transabdominally for all women. The involution process of the uterus was assessed by measuring the anteroposterior diameter and uterine cavity. The appearance of the uterine cavity contents was documented. Factors related to the involution process: parity, breast-feeding, smoking and infant's birth weight were also evaluated. RESULTS: The maximum anteroposterior diameter of the uterus diminished substantially and progressively from 93 mm on day one postpartum to 38.5 mm on day 56. The maximum anteroposterior diameter of the uterine cavity diminished from 15.2 mm on day one to 4.0 mm on day 56. The position of the uterus, its shape and the appearance of its cavity during the normal puerperium were observed. The uterus was most often retroverted and empty in the early puerperium. Fluid and debris in the whole cavity were seen in mid puerperium. In late puerperium, the cavity was empty and appeared as a thin white line. No correlation was found between the involution of the uterus and parity, breast-feeding and the infant's birth weight. CONCLUSION: Transabdominal sonography is suitable for examination of the uterus during the early puerperium period. The uterine body and position, as well as the cavity, are easy to examine by ultrasound. Accumulation of fluid and debris in the uterine cavity is a common and insignificant finding of the involuting uterus. PMID- 14758378 TI - The effect of Ramadan fasting on amniotic fluid volume. AB - OBJECTIVE: This cross-sectional study was carried out to assess the influence of Ramadan fasting on amniotic fluid volume. METHODS: For this purpose 2 groups of fasting (n=28) and non-fasting (n=25) pregnant women were investigated for amniotic fluid index (AFI) and deepest vertical pocket of amniotic fluid in November (Ramadan month) of 2001-2002 in Kerman Prenatal Clinics in Kerman, Iran. Mean of gestational age based on the last menstrual period and ultrasound reports were 29.6 +/- 5.8 week and 29.5 +/- 4.2 week in fasting and non-fasting groups. RESULTS: There was no significant difference between the 2 groups in age, gestational age, gravidity, parity and systolic/diastolic blood pressures. The mean deepest vertical pocket in fasting and non-fasting groups were 65.9 +/- 12.9 mm and 62.7 +/- 6.5 mm and there was no significant difference between the 2 groups in this regard. Mean AFI in fasting group (189.9 +/- 35.9 mm) and in non fasting group (166.8 +/- 25.3 mm) showed a significant difference (p less than 0.05). CONCLUSION: Based on the results of the present study fasting in Ramadan has no significant effect on the decrease of AFI, deepest vertical pocket and amniotic fluid volume. PMID- 14758379 TI - The effect of fasting in Ramadan on patients with heart disease. AB - OBJECTIVE: To determine the clinical and biochemical effects of fasting during Ramadan on patients with cardiac disease. METHODS: Eighty-six outpatients with heart disease with intention to fast were studied in the month of Ramadan 1996 (1416 H) at the King Fahd Armed Forces Hospital, Jeddah, Kingdom of Saudi Arabia. Detailed clinical and biochemical assessments were performed within 3 days before the start of Ramadan and then on the last day of Ramadan. RESULTS: There were 54 (62.8%) males and 32 (37.2%) females with a mean age of 56.3 years (range, 17-84 ). Forty-six patients (53%) had coronary artery disease, 23 patients (27%) had valvular heart disease, 13 patients (15%) had congestive heart failure and 4 patients (5%) were treated for arrhythmia. Sixty-two patients (72%) were in New York Heart Association (NYHA) Class I, 18 patients (21%) in Class II, and 6 patients (7%) were in Class III. Seventy-four patients (86%) managed to fast during the entire Ramadan, 9 patients (10.4%) missed the fasting for up to 7 days, and 3 patients (3.5%) could not fast. There were no significant changes in the NYHA Class (p=0.12). No significant changes occurred in any of the hematological or biochemical parameters during the fasting of Ramadan. CONCLUSION: The effects of fasting during Ramadan on stable patients with cardiac disease are minimal. The majority of patients with stable cardiac disease can fast during Ramadan without significant detrimental effects. PMID- 14758380 TI - Prevalence of renal artery stenosis in patients undergoing routine cardiac catheterization. AB - OBJECTIVE: To determine the prevalence of renal artery stenosis (RAS) and associated risk factors in patients undergoing cardiac catheterization for suspected coronary artery disease. METHODS: Three hundred and fifty-four consecutive patients (71 female) were studied at the Cardiology Unit of King Abdullah University Teaching Hospital, Irbid, Jordan, between May 2002 and May 2003. Left-sided cardiac catheterization and abdominal aortography were performed to screen for coronary and renal artery disease. RESULTS: Of the 354 patients, 285 had coronary artery disease and 27 had RAS. Significant RAS was present in 11 patients. Patients with RAS were older (66 +/- 8 versus 59 +/- 10, mean +/- SD; p=0.004), had higher incidence of systolic hypertension (156 +/-14 versus 130 +/- 16 mm Hg; p=0.005), diabetes mellitus (72% versus 38%; p=0.004), smoker (85% versus 55; p=0.005), and had less than 2 coronary lesions. CONCLUSION: The prevalence of significant and insignificant RAS is 3.1% and 4.5%. Diagnostic yield increase in elderly patient with less than 2 coronary lesions, elevated systolic pressure, smoking, diabetes mellitus, and electrocardiogram criteria of left ventricular hypertrophy. PMID- 14758382 TI - Mutation analysis of the breast cancer gene BRCA1 among breast cancer Jordanian females. AB - OBJECTIVE: To screen mutations of the tumor suppressor breast cancer susceptibility gene 1 (BRCA1) within 3 exons among Jordanian breast cancer females. METHODS: A total of 135 Jordanian breast cancer females were genetically analyzed by denaturing gradient electrophoresis (DGGE) for mutation detection in 3 BRCA1 exons (2, 11 and 20) between 2000-2002 in Al-Basheer Hospital, Amman, Jordan. RESULTS: Of the studied patients 50 had a family history of breast cancer, 28 had a family history of cancer other than breast cancer, and 57 had no family history of any cancer. Five germline mutations were detected among breast cancer females with a family history of breast cancers (one in exon 2 and 4 mutations in exon 11). Another germline mutation (within exon 11) was detected among breast cancer females with family history of cancer other than breast cancer, and no mutation was detected among breast cancer females with no family history of any cancer or among normal control females. CONCLUSION: Screening mutations within exon 2, exon 11 and exon 20 showed that most screened mutations were within BRCA1 exon 11 among breast cancer Jordanian families with a family history of breast cancer. PMID- 14758381 TI - Thyroid cancer in Yemen. AB - OBJECTIVE: To evaluate the characteristics of thyroid cancer (TC) patients in regard to demographic distribution, histological variants, mode of presentation and modalities of diagnosis and treatment. METHODS: We retrospectively audited the records of 97 consecutive cases with histologically proved thyroid cancer between 1997 and 2001 presenting to the Kuwait University Hospital, Sana'a, Yemen. RESULTS: Patients with TC in this study comprise 17.7% of goiter patients who were admitted at the same period. Females constituted 89.7% (n=87), and males 10.3% (n=10). The average age of diagnosis was 38.4 years. More than two thirds were at the age of <40. Among patients with goiter, the percentage of carcinoma was higher in the following groups, males (37.9% versus 16.6%, p=0.028), patients aged >47 (25.8% versus 12.6%, p=0.001), patients with enlarged lymph nodes (9.3% versus 3.8%, p=0.020), and patients with recurrent disease after being operated for a presumably benign disease (8.2% versus 2.9%, p=0.012). The average period since patients noticed the swelling until seeking medical help was 4 years. Most patients (90%) came from highland areas. Multinodular swelling was the most common clinical finding (43.3%), and hoarseness was the most common symptom (17.5%). In histopathological examination, papillary carcinoma accounted for 93.8% of the cases and papillary microcarcinoma was found in 10 cases (10.3%). The papillary/follicular carcinoma ratio was 22:1. Hormone assay and ultrasonic imaging were the most commonly used investigations. Sub total thyroidectomy was the most common procedure used in treatment (39%). CONCLUSION: Not all histological variants of TC are represented in this study. Papillary carcinoma formed the bulk of TC cases. Salt iodization program might have an effect on the incidence of thyroid malignancy, and on the papillary/follicular carcinoma ratio. Better level of expertise is needed in the field of fine needle aspiration and ultrasonography. A consensus has to be reached, which is based on our environment and capabilities, where TC has to be managed aggressively by experienced surgeons. Yemen is in real need of a national cancer registry to assess the problem on a national level. PMID- 14758383 TI - Scorpion sting syndrome in a general hospital in Saudi Arabia. AB - OBJECTIVE: To evaluate the incidence of scorpion stings and to draw the attention of clinicians, concerning the dilemma of scorpion sting syndrome and its management in the Kingdom of Saudi Arabia (KSA). METHODS: In a retrospective survey, 251 cases were presented with scorpion stings to the Emergency Department of the Armed Forces Hospital, Riyadh, KSA during a period of 15 years (April 1986 to April 2000). Their ages ranged from 2 months to 101 years (male to female ratio was 2.6:1) and 70.6% of the cases were under 20 years of age. RESULTS: The mean annual incidence of the scorpion sting in the month of May was 16.7%, with the highest mean percentage of approximately 36% each year. The seasonal sting cycle showed the highest record was in the summer period (51%) as compared to the lowest winter period (10%). Local pain was the primary presenting complaint (95%), with a total systemic toxicity of 78.3% and 35.2% of these were children. Hypertension, sweating, salivation and tachycardia were the most common signs of systemic symptoms. The majority of patients received analgesia, local anesthetics, and application of ice and a period of observation dictated by clinical findings. Eighty-two patients (32.6%) had signs of serious envenoming, requiring admission. There were no deaths. All of these cases were mainly treated symptomatically. CONCLUSION: The beneficial effect of antivenom in protecting victims against scorpion stings is still questionable. The higher risk groups of systemic toxicity were either those with ages less than 10 years or greater than 50 years, being more susceptible due to their decreased physiologic reserves and increased debilitation. PMID- 14758384 TI - The Watched Structure Clinical Examination (WASCE) as a tool of assessment. AB - OBJECTIVE: Assessment of medical students' clinical competencies is still evolving. The aim of this study was to find out the effectiveness of a new tool of assessment to assess medical students at the end of clinical rotations. METHODS: A new tool has been developed in the College of Medicine and Medical Sciences, Arabian Gulf University, Manama, Bahrain, called the Watched Structure Clinical Examination (WASCE). It was used at the end of the Family Medicine clinical rotation during the academic year 2000-2001 involving 62 final year students. RESULTS: The study found a significant statistical correlation between the students' results in the WASCE and their results in Doctor of Medicine final examination, which included the written examination, the patient encountered clinical examination and the Objective Structured Clinical Examination (OSCE). CONCLUSION: Watched Structured Clinical Examination can be a useful method of assessment for examining certain clinical skills, with an advantage over the OSCE in that it is less time consuming, more cost effective, requires less supervising staff to conduct the examination and, more importantly, it is less stressful to the students. PMID- 14758385 TI - Effects of short term metformin administration on androgens in diabetic men. AB - OBJECTIVE: Metformin, an oral hypoglycemic agent, has several other metabolic and hormonal effects. This study aims at identifying the metabolic effect of metformin on androgens in diabetic men. METHODS: The study was conducted at The National Center for Diabetes Endocrinology and Genetics, Jordan University Hospital, Amman, Jordan from April 2001 to September 2001. We studied 15 men with type 2 diabetes mellitus by measuring fasting serum glucose, insulin, glycosylated hemoglobin, total and free testosterone, sex hormone binding globulin, dehydroepiandrosterone sulphate, 17-OH progesterone, luteinizing hormone, and follicle stimulating hormone before and after a short course of metformin. RESULTS: There was a significant decrease in fasting serum glucose and glycosylated hemoglobin and increase in the level of 17-OH progesterone. The remainder of the measured parameters did not show any significant change. Although serum glucose and glycosylated hemoglobin decreased insulin levels were not changed. CONCLUSION: In contrast to normal men there was no change in androgen levels in diabetics but the 17-OH progesterone was elevated. PMID- 14758386 TI - Dementia in Qatar. AB - OBJECTIVE: Dementia is a major public health problem among the elderly in developed countries and a growing problem in the underdeveloped countries. There are no published data on dementia in any of the Arabic countries. The aim of this study was to determine the different sub-types of dementia among Qataris. METHODS: A retrospective and prospective ongoing hospital-based study in which all medical records of the patients with diagnosis of dementia seen at the Hamad General Hospital, Doha, Qatar, between June 1997 and June 2003, whether inpatient or outpatient were reviewed. Dementia was defined according to diagnostic and statistical manual (DSM) IV criteria. Those who had dementia were evaluated by a psychologist, psychiatrist, neurologist and a geriatrician. All had brain computerized tomography, magnetic resonance imaging or both and routine blood tests. Finally, they were classified into sub-types according to the cause of dementia. RESULTS: Out of 300 patients, 134 fulfilled the inclusion criteria, most of them were illiterate, married and non-smokers. Among those dementia sub types were: Alzheimer disease (AD) 39 (29%), vascular dementia (VaD) 30 (22%), mixed AD and VaD 20 (15%) and Parkinson's disease with dementia 8 (6%) were noted. The rest of them had dementia due to other medical conditions. CONCLUSION: Our study showed that AD is more prevalent than VaD. It also showed that patients and their families seek medical help late due to the general belief among the public that forgetfulness and other associated cognitive impairment are part of the normal aging process. The emergence of new drugs and advancement in the prevention of cerebrovascular diseases makes early diagnosis of dementia sub-type important. A community based study, to show the real prevalence and incidence of sub-types of dementia, is highly indicated. These data are necessary for planning and setting up community services and health care programs for demented patients. PMID- 14758387 TI - The Arabic version of childhood health assessment questionnaire modified for Arabic children. AB - OBJECTIVE: To determine the feasibility, reliability and validity of the childhood health assessment questionnaire--modified for Arab children (CHAQ- MAC). METHODS: One hundred and eighteen modified questionnaires were completed by 75 juvenile rheumatoid arthritis (JRA) patients and their parents attending the Pediatric Rheumatology Clinic at the King Faisal Specialist Hospital and Research Centre, Riyadh, Kingdom of Saudi Arabia over an 18 month period (January 1996 to May 1997). RESULTS: The modified questionnaire was self-administered by 82% of the parents. The median time to complete the questionnaire was 10 minutes. The main difficulty in comprehension was discomfort dimension (visual analogue scale [VAS] and morning stiffness). Test retest reliability was good (r=0.79). Validity of the CHAQ-MAC was confirmed by the strong correlation between disability index and VAS score (r=0.58). Functional activities that caused the most difficulties were cross sitting, assuming the prayer position, and using the Arabic style toilet. CONCLUSION: The modified CHAQ is a suitable assessment tool for Arab children suffering from JRA. PMID- 14758388 TI - Effect of elevated-rim acetabular liner and 32-mm femoral head on stability in total hip arthroplasty. AB - OBJECTIVE: Although the theoretical attractions of the elevated rim are obvious and have been widely accepted as a mean to improve the postoperative stability, the clinical advantages have not been demonstrated. The aim of this study is to further evaluate the elevated liners contribution to stability. METHODS: Forty six patients with 50 hips undergoing primary total hip arthroplasty (THA) were enrolled in this study, conducted in Rush Hospital, Chicago, Illinois, United State of America, between March 2001 and February 2003. We tried to determine the amount of additional stability that can be provided by elevated-rim liner compared to the non-elevated liners and the stability of the hip with a 32 mm femoral head compared to 28 mm head. RESULTS: Our results showed that a 10 degree elevated-rim acetabular liners increased hip stability by an additional 8.2 degrees of internal rotation. The 32 mm head provided 7.3 degrees of internal rotation. The increases were statistically significant (p value is less than 0.0001). CONCLUSION: The findings of this study clearly show that an elevated-rim liner, and independently the 32 mm head, may contribute to hip stability. PMID- 14758389 TI - Wandering spleen. AB - Congenital malformations of the spleen are rare. We report 3 cases of wandering spleen presented as abdominal or pelvi-abdominal mass. Two patients were suffering from chronic lower abdominal pain with thrombosed splenic pedicle and the third patient had an acute abdomen. All patients underwent splenectomies. Abdominal ultrasound, computerized tomography, Doppler ultrasound, and radioisotope studies were used to confirm the diagnosis. The clinical, diagnostic and treatment modalities are discussed. PMID- 14758390 TI - Hemorrhagic episodes in hemophiliacs simulating abdominal surgical emergencies. AB - Hemophiliacs are subjected to develop episodes of spontaneous bleeding at different sites of the body, primarily the knees. On occasions, such episodes affect the abdomen. The picture engendered in such cases may mimic that of an abdominal emergency requiring surgical intervention. Such ill advised and unwarranted intervention may end with the patient's death. With the proper employment of radiology, the correct diagnosis may be reached and consequently, conservative treatment, in which factor VIII plays the major role, instituted. Here, we describe the clinical course of 2 patients with hemophilia A who suffered bleeding in the abdomen and were treated conservatively with a successful outcome. PMID- 14758391 TI - Choledochal cyst. AB - Choledochal cyst, which is characterized by dilatation of the biliary ducts, is common in Asian countries, mainly Japan, but relatively rare worldwide. This report describes 2 Saudi female children with choledochal cysts, with emphasis on long term follow-up. PMID- 14758392 TI - Amlodipine associated hyperpigmentation. AB - Amlodipine is a potent peripheral and coronary vasodilator with high selectivity for vascular smooth muscle, and is widely used in mild to moderate hypertension, chronic stable angina and vasospastic angina. Its most prevalent side effects are peripheral edema, flushing and headache. Cutaneous adverse reactions associated with amlodipine have been rarely reported. Herein, a male patient is described to develop oral mucosal and cutaneous hyperpigmentation one year after starting amlodipine, which became more noticeable with time. Although cutaneous hyperpigmentation was most prominent on the photoexposed areas, there was no history of previous photosensitivity, pruritus or flushing. To our knowledge, no case of oral and cutaneous hyperpigmentation associated with amlodipine has been formally reported up to date. PMID- 14758393 TI - Thrombocytopenia responding to red blood cell transfusion. AB - Three patients with severe symptomatic iron deficiency anemia and thrombocytopenia had a significant rise in platelet count a few days following packed red blood cell transfusion. Pre-transfusion platelet count of patient one was 17 x 10(9)/L, 22 x 10(9)/L in patient 2 and 29 x 10(9)/L in patient 3. On the 6th day post transfusion, the platelet count rose to 166 x 10(9)/L in patient one, 830 x 10(9)/L in patient 2 and 136 x 10(9)/L in patient 3. The possible mechanisms behind such an unreported observation are discussed. PMID- 14758395 TI - Ramsay Hunt syndrome in focal segmental glomerulosclerosis. PMID- 14758396 TI - Candida guilliermondii fungemia. To treat or not to treat. PMID- 14758397 TI - Acute chest syndrome in sickle cell disease. PMID- 14758398 TI - Spectrum of childhood poisoning in a tertiary center in the Eastern Saudi Arabia. PMID- 14758399 TI - C-reactive protein in diabetes mellitus and hyperlipidemia. PMID- 14758400 TI - Risk of morbid obesity with pregnancy. PMID- 14758401 TI - Bowel anastomosis. A comparative study of various surgical techniques. PMID- 14758402 TI - Using knowledge management to make health systems work. PMID- 14758403 TI - Health expectancy in the Russian Federation: a new perspective on the health divide in Europe. AB - OBJECTIVES: To compare life expectancy and healthy life expectancy in the Russian Federation and in countries of Eastern and Western Europe. METHODS: WHO mortality data and data on self-reported health from the World Values Survey and the Russian Longitudinal Monitoring Survey were used to compare the above three regions. Life expectancy was calculated using Sullivan's method, with years of life lived divided into healthy and unhealthy. The gap in healthy life expectancy between the Russian Federation and Western Europe was examined by decomposing the difference by gender and age. FINDINGS: The probability of remaining alive and healthy declines faster in the Russian Federation than in Western Europe, with the gap between Eastern Europe and the Russian Federation widening at older ages. In the Russian Federation, this rapid decline is due mainly to the high probability of death or of poor health for men and women, respectively. CONCLUSIONS: There is a large toll of premature male mortality in the Russian Federation but there also appears to be a substantial burden of ill-health among women. As in other countries, the responses of men and women to adversity differ, leading to premature death in men but survival in a poor state of health in women. Epidemiological studies including objective measures of health would help policy-makers to estimate more precisely the scale and nature of this problem. Policy-makers must recognize that health expectancy in the Russian Federation is reduced in both men and women. PMID- 14758404 TI - Towards valid and comparable measurement of population health. PMID- 14758405 TI - Influence of pesticide regulation on acute poisoning deaths in Sri Lanka. AB - OBJECTIVES: To assess in a developing Asian country the impact of pesticide regulation on the number of deaths from poisoning. These regulations, which were implemented in Sri Lanka from the 1970s, aimed to reduce the number of deaths - the majority from self-poisoning - by limiting the availability and use of highly toxic pesticides. METHODS: Information on legislative changes was obtained from the Ministry of Agriculture, national and district hospital admission data were obtained from the Sri Lanka Health Statistics Unit, and individual details of deaths by pesticide poisoning were obtained from a manual review of patients' notes and intensive care unit records in Anuradhapura. FINDINGS: Between 1986 and 2000, the total national number of admissions due to poisoning doubled, and admissions due to pesticide poisoning increased by more than 50%. At the same time, the case fatality proportion (CFP) fell for total poisonings and for poisonings due to pesticides. In 1991_92, 72% of pesticide-induced deaths in Anuradhapura were caused by organophosphorus (OP) and carbamate pesticides - in particular, the WHO class I OPs monocrotophos and methamidophos. From 1991, the import of these pesticides was reduced gradually until they were banned for routine use in January 1995, with a corresponding fall in deaths. Unfortunately, their place in agricultural practice was taken by the WHO class II organochlorine endosulfan, which led to a rise in deaths from status epilepticus - from one in 1994 to 50 in 1998. Endosulfan was banned in 1998, and over the following three years the number of endosulfan deaths fell to three. However, at the end of the decade, the number of deaths from pesticides was at a similar level to that of 1991, with WHO class II OPs causing the most deaths. Although these drugs are less toxic than class I OPs, the management of class II OPs remains difficult because they are, nevertheless, still highly toxic, and their toxicity is exacerbated by the paucity of available facilities. CONCLUSION: The fall in CFP amidst a rising incidence of self-poisoning suggests that Sri Lanka's programmes of pesticide regulation were beneficial. However, a closer inspection of pesticide-induced deaths in one hospital revealed switching to other highly toxic pesticides, as one was banned and replaced in agricultural practice by another. Future regulation must predict this switching and bear in mind the ease of treatment of replacement pesticides. Furthermore, such regulations must be implemented alongside other strategies, such as integrated pest management, to reduce the overall pesticide availability for self-harm. PMID- 14758406 TI - Burden of tuberculosis in Kampala, Uganda. AB - OBJECTIVE: To determine the prevalence and incidence of tuberculosis in one of Uganda's poor peri-urban areas. METHODS: Multi-stage sampling was used to select a sample of households whose members were evaluated for presence of signs and/or symptoms of active tuberculosis; history of tuberculosis treatment; and relevant demographic, socioeconomic, and household environment characteristics. Patients with suspected tuberculosis underwent standardized evaluation for active disease. FINDINGS: A sample of 263 households with 1142 individuals was evaluated. Nineteen people were classified as having had tuberculosis during the one-year reference period (May 2001-April 2002): nine (47%) cases already had been diagnosed through the health care system, while 10 cases (53%) were diagnosed through the survey. The prevalences for all forms of tuberculosis and for sputum smear-positive tuberculosis were 14.0 (95% confidence interval (CI) 7.8-20.3) and 4.4 (CI = 0.83-7.89) per thousand, respectively. The incidences for all forms of tuberculosis and for sputum smear-positive tuberculosis were 9.2 (CI = 3.97-14.4) and 3.7 (CI = 0.39-6.95) per thousand per year, respectively. CONCLUSION: The rate of tuberculosis in this peri-urban community was exceptionally high and may be underestimated by current surveillance systems. The need for interventions aimed at reducing tuberculosis transmission in this, and other similar communities with high case rates, is urgent. PMID- 14758407 TI - Training pharmacy workers in recognition, management, and prevention of STDs: district-randomized controlled trial. AB - OBJECTIVES: To determine the effectiveness of an intervention for pharmacy workers in improving their recognition and management of sexually transmitted disease (STD) syndromes. METHODS: We randomly selected 14 districts (total population nearly 4 million) from the 24 districts of low socioeconomic status in Lima, Peru. We randomly assigned paired districts to receive training and support for management and prevention of STDs or a control intervention about management of diarrhoea. The STD intervention included interactive luncheon seminars on recognition and management of four STD syndromes (urethral discharge, vaginal discharge, genital ulcers, and pelvic inflammatory disease) and STD/HIV prevention counselling; monthly pharmacy visits by "prevention salespersons" who distributed materials that included "STD/HIV prevention packets" containing information, condoms, and cards given to patients for referral of their sex partners; and workshops for physicians on managing patients with STD syndromes referred from pharmacies. Standardized simulated patients visited pharmacies in intervention and control districts at one, three, and six months after training to assess outcomes. FINDINGS: Standardized simulated patients reported significantly better recognition and management (appropriate antimicrobial regimens provided for discharge syndromes and referral to specially trained physicians for genital ulcers or pelvic inflammatory disease) by pharmacy workers of all four STD syndromes. They also reported significantly more frequent recommendations for use of condoms and treatment of partners at pharmacies in intervention districts than in control districts (by "intention-to-train" analyses, P<0.05 for 47/48 primary outcome comparisons). CONCLUSION: Training was feasible and effectively improved pharmacy workers' practices. PMID- 14758408 TI - Knowledge for better health: a conceptual framework and foundation for health research systems. AB - Health research generates knowledge that can be utilized to improve health system performance and, ultimately, health and health equity. We propose a conceptual framework for health research systems (HRSs) that defines their boundaries, components, goals, and functions. The framework adopts a systems perspective towards HRSs and serves as a foundation for constructing a practical approach to describe and analyse HRSs. The analysis of HRSs should, in turn, provide a better understanding of how research contributes to gains in health and health equity. In this framework, the intrinsic goals of the HRS are the advancement of scientific knowledge and the utilization of knowledge to improve health and health equity. Its four principal functions are stewardship, financing, creating and sustaining resources, and producing and using research. The framework, as it is applied in consultation with countries, will provide countries and donor agencies with relevant inputs to policies and strategies for strengthening HRSs and using knowledge for better health. PMID- 14758409 TI - Do childhood vaccines have non-specific effects on mortality? AB - A recent article by Kristensen et al. suggested that measles vaccine and bacille Calmette-Gu rin (BCG) vaccine might reduce mortality beyond what is expected simply from protection against measles and tuberculosis. Previous reviews of the potential effects of childhood vaccines on mortality have not considered methodological features of reviewed studies. Methodological considerations play an especially important role in observational assessments, in which selection factors for vaccination may be difficult to ascertain. We reviewed 782 English language articles on vaccines and childhood mortality and found only a few whose design met the criteria for methodological rigor. The data reviewed suggest that measles vaccine delivers its promised reduction in mortality, but there is insufficient evidence to suggest a mortality benefit above that caused by its effect on measles disease and its sequelae. Our review of the available data in the literature reinforces how difficult answering these considerations has been and how important study design will be in determining the effect of specific vaccines on all-cause mortality. PMID- 14758410 TI - Exercise interventions: defusing the world's osteoporosis time bomb. AB - Osteoporosis is a major public health problem, affecting millions of people worldwide. The associated health care costs are growing in parallel with increases in elderly populations, and it is expected that the number of osteoporotic fractures will double over the next 50 years. The best way to address osteoporosis is prevention. Some interventions to maximize and preserve bone mass have multiple health benefits and are cost-effective. For example, modifications to diet and lifestyle can help to prevent osteoporosis, and could potentially lead to a significant decrease in fracture rates; and exercise is a valuable adjunct to programmes aimed at alleviating the risks and symptoms of osteoporosis. Practising exercise at a young age helps maximize the mineral density of bones while they are still growing and maturing, and continuing to excercise minimizes bone loss later in life. Not only does exercise improve bone health, it also increases muscle strength, coordination, balance, flexibility and leads to better overall health. Walking, aerobic exercise, and t'ai chi are the best forms of exercise to stimulate bone formation and strengthen the muscles that help support bones. Encouraging physical activity at all ages is therefore a top priority to prevent osteoporosis. PMID- 14758411 TI - Mortality in relation to economic development. PMID- 14758412 TI - The changing relation between mortality and level of economic development. 1975. PMID- 14758413 TI - Food safety: the fourth pillar in the strategy to prevent infant diarrhoea. PMID- 14758414 TI - Medical spending and health outcome in Nepal: problems with technology or its distribution? PMID- 14758415 TI - Exposures to lead require ongoing vigilance. PMID- 14758416 TI - Iraq health minister plans future Iraqi health system. PMID- 14758417 TI - Expert Committee finds little fault in Hong Kong's response to SARS. PMID- 14758418 TI - Malnutrition leading cause of death in post-war Angola. PMID- 14758419 TI - Nations fail to agree on extent of human cloning ban. PMID- 14758420 TI - Developing countries overstate vaccination coverage. PMID- 14758421 TI - WHO launches guidelines to speed up delivery of antiretroviral drugs to HIV/AIDS patients. PMID- 14758422 TI - Workforce crisis a major obstacle in global tuberculosis control. PMID- 14758423 TI - Death of humanitarian worker major loss for tuberculosis control in Africa. PMID- 14758424 TI - WHO launches blindness prevention "Tool Kit". PMID- 14758425 TI - Global burden of musculoskeletal disease revealed in new WHO report. PMID- 14758426 TI - WHO manual on child health awarded the 2003 Prescrire Prize. PMID- 14758427 TI - AIDS as a global emergency. PMID- 14758428 TI - Equitable access to scientific and technical information for health. PMID- 14758429 TI - Dried venous blood samples for the detection and quantification of measles IgG using a commercial enzyme immunoassay. AB - OBJECTIVES: To determine whether samples of dried venous blood (DVB) were an acceptable alternative to serum for detecting measles-specific IgG in a commercial enzyme immunoassay. METHODS: Paired samples of serum and DVB were collected from 98 suspected cases of measles and 1153 schoolchildren in Victoria, Australia. All samples were tested using the Dade Behring Enzygnost Anti-Measles Virus/IgG immunoassay. DVB samples were eluted using either the sample buffer provided with the kit or 5% dry milk powder in phosphate-buffered saline-Tween 20. FINDINGS: DVB samples eluted by sample buffer showed significantly better linear correlation to the serum samples than did DVB samples eluted in 5% dry milk in phosphate-buffered saline-Tween 20. To improve the comparability of serum and DVB samples an adjustment factor of 1.28 was applied to the optical density (OD) values of DVB. This adjustment also enabled quantification of the titre of measles IgG in mIU/ml directly from the OD value using the alpha calculation as specified by the kit protocol. For DVB samples stored for less than six months at 4 degrees C, the assay showed an overall sensitivity of 98.4% and a specificity of 97.2% compared with the results of serum testing. CONCLUSION: These results illustrate the potential for DVB samples to be widely used with the Dade Behring enzyme immunoassay system for determining the immunity of the individual and the population to the measles virus. PMID- 14758430 TI - Integrated Management of Childhood Illnesses strategy: compliance with referral and follow-up recommendations in Gezira State, Sudan. AB - OBJECTIVES: To determine the extent to which families follow referral and follow up recommendations given in accordance with the Integrated Management of Childhood Illnesses (IMCI) strategy and the factors that influence families' responses to such recommendations. METHODS: Children aged 2 months-5 years who presented to an IMCI-trained health worker in Massalamia Health Area, Sudan, were recruited. Children with an IMCI classification that indicated the need for referral or follow-up were traced to determine whether the family complied with the referral or follow-up recommendation. Caretakers were interviewed to find out why they had or had not complied. Focus group discussions were held with health workers, caretakers, and community members. FINDINGS: Overall, 5745 children were enrolled. Of these, 162 (3%) were considered to be in need of urgent referral: 53 (33%) attended a hospital on the day of the referral, with a further 37 (23%) visiting the hospital later than the day of referral. About half of families cited cost as the reason for not visiting a hospital. A total of 1197 (21%) children were classified as needing follow-up. Compliance with a follow-up recommendation was 44% (529 children). Almost 165 (90%) of caretakers who were aware of and did not comply with follow-up, said they had not done so because the child was better. Compliance increased with the caretaker's level of education, if drugs were provided during the first visit, and if the follow-up period was short (2 or 5 days). CONCLUSION: In Massalamia--a resource-constrained environment in which IMCI implementation was well received by the community--only about half of children judged to be in need of urgent referral were taken for that care within 24 hours. Most children in need of follow-up received their first treatment dose in the health facility. This aspect of IMCI was commented upon favourably by caretakers, and it may encourage them to return for follow-up. Rates of return might also improve if return visits for children currently asked to return after 14 or 30 days were scheduled earlier. PMID- 14758431 TI - Risk factors for early infant mortality in Sarlahi district, Nepal. AB - OBJECTIVES: Early infant mortality has not declined as rapidly as child mortality in many countries. Identification of risk factors for early infant mortality may help inform the design of intervention strategies. METHODS: Over the period 1994 97, 15,469 live-born, singleton infants in rural Nepal were followed to 24 weeks of age to identify risk factors for mortality within 0-7 days, 8-28 days, and 4 24 weeks after the birth. FINDINGS: In multivariate models, maternal and paternal education reduced mortality between 4 and 24 weeks only: odds ratios (OR) 0.28 (95% confidence interval (CI) = 0.12-0.66) and 0.63 (95% CI = 0.44-0.88), respectively. Miscarriage in the previous pregnancy predicted mortality in the first week of life (OR = 1.98, 95% CI = 1.37-2.87), whereas prior child deaths increased the risk of post-neonatal death (OR = 1.85, 95% CI 1.24-2.75). A larger maternal mid-upper arm circumference reduced the risk of infant death during the first week of life (OR = 0.88, 95% CI = 0.81-0.95). Infants of women who did not receive any tetanus vaccinations during pregnancy or who had severe illness during the third trimester were more likely to die in the neonatal period. Maternal mortality was strongly associated with infant mortality (OR = 6.43, 95% CI = 2.35-17.56 at 0-7 days; OR = 11.73, 95% CI = 3.82-36.00 at 8-28 days; and OR = 51.68, 95% CI = 20.26-131.80 at 4-24 weeks). CONCLUSION: Risk factors for early infant mortality varied with the age of the infant. Factors amenable to intervention included efforts aimed at maternal morbidity and mortality and increased arm circumference during pregnancy. PMID- 14758432 TI - Single-dose versus multi-dose vaccine vials for immunization programmes in developing countries. AB - Excessive vaccine wastage and safety concerns have prompted the international health community to develop and supply vaccines in formats other than the standard multi-dose vial. This article presents a programmatic and economic comparison of the major differences between the multi-dose vials and single-dose formats used for immunization services in developing countries. Multi-dose vials, in general, sell at a lower per-dose price and occupy less cold-chain capacity than single-dose formats. However, higher wastage rates may offset these benefits, especially for more expensive vaccines. Single-dose formats offer several important programmatic benefits, such as increased vaccination opportunities and improved vaccine safety. One single-dose format, the prefilled auto-disable (AD) device, provides additional injection safety and convenience features because it physically combines the vaccine and AD syringe. Selecting the appropriate vaccine presentation will depend on many factors. However, multi-dose vials are likely to be most appropriate for cheaper vaccines and in settings where cold-chain storage capacity is restricted. Single-dose formats will be most appropriate for more expensive vaccines and where there are problems with unsafe injection practices. Prefilled AD injection devices will be particularly useful in expanding outreach services while eliminating the possibility of needle reuse. PMID- 14758433 TI - Meta-analysis of residential exposure to radon gas and lung cancer. AB - OBJECTIVES: To investigate the relation between residential exposure to radon and lung cancer. METHODS: A literature search was performed using Medline and other sources. The quality of studies was assessed. Adjusted odds ratios with 95% confidence intervals (CI) for the risk of lung cancer among categories of levels of exposure to radon were extracted. For each study, a weighted log-linear regression analysis of the adjusted odds ratios was performed according to radon concentration. The random effect model was used to combine values from single studies. Separate meta-analyses were performed on results from studies grouped with similar characteristics or with quality scores above or equal to the median. FINDINGS: Seventeen case-control studies were included in the meta-analysis. Quality scoring for individual studies ranged from 0.45 to 0.77 (median, 0.64). Meta-analysis based on exposure at 150 Bq/m3 gave a pooled odds ratio estimate of 1.24 (95% CI, 1.11-1.38), which indicated a potential effect of residential exposure to radon on the risk of lung cancer. Pooled estimates of fitted odds ratios at several levels of randon exposure were all significantly different from unity--ranging from 1.07 at 50 Bq/m3 to 1.43 at 250 Bq/m3. No remarkable differences from the baseline analysis were found for odds ratios from sensitivity analyses of studies in which > 75% of eligible cases were recruited (1.12, 1.00-1.25) and studies that included only women (1.29, 1.04-1.60). CONCLUSION: Although no definitive conclusions may be drawn, our results suggest a dose-response relation between residential exposure to radon and the risk of lung cancer. They support the need to develop strategies to reduce human exposure to radon. PMID- 14758434 TI - Coverage of pilot parenteral vaccination campaign against canine rabies in N'Djamena, Chad. AB - Canine rabies, and thus human exposure to rabies, can be controlled through mass vaccination of the animal reservoir if dog owners are willing to cooperate. Inaccessible, ownerless dogs, however, reduce the vaccination coverage achieved in parenteral campaigns. This study aimed to estimate the vaccination coverage in dogs in three study zones of N'Djamena, Chad, after a pilot free parenteral mass vaccination campaign against rabies. We used a capture-mark-recapture approach for population estimates, with a Bayesian, Markov chain, Monte Carlo method to estimate the total number of owned dogs, and the ratio of ownerless to owned dogs to calculate vaccination coverage. When we took into account ownerless dogs, the vaccination coverage in the dog populations was 87% (95% confidence interval (CI), 84-89%) in study zone I, 71% (95% CI, 64-76%) in zone II, and 64% (95% CI, 58-71%) in zone III. The proportions of ownerless dogs to owned dogs were 1.1% (95% CI, 0-3.1%), 7.6% (95% CI, 0.7-16.5%), and 10.6% (95% CI, 1.6-19.1%) in the three study zones, respectively. Vaccination coverage in the three populations of owned dogs was 88% (95% CI, 84-92%) in zone I, 76% (95% CI, 71-81%) in zone II, and 70% (95% CI, 66-76%) in zone III. Participation of dog owners in the free campaign was high, and the number of inaccessible ownerless dogs was low. High levels of vaccination coverage could be achieved with parenteral mass vaccination. Regular parenteral vaccination campaigns to cover all of N'Djamena should be considered as an ethical way of preventing human rabies when post exposure treatment is of limited availability and high in cost. PMID- 14758435 TI - Meningococcal meningitis in sub-Saharan Africa: the case for mass and routine vaccination with available polysaccharide vaccines. AB - Endemic and epidemic group A meningococcal meningitis remains a major cause of morbidity and mortality in sub-Saharan Africa, despite the availability of the safe and inexpensive group A meningococcal polysaccharide vaccine, which is protective at all ages when administered as directed. Despite optimal therapy, meningococcal meningitis has a 10% fatality rate and at least 15% central nervous system damage. WHO's policy of epidemic containment prevents, at best, about 50% of cases and ignores endemic meningitis, which is estimated at 50,000 cases per year. The effectiveness of group A, C, W135, and Y capsular polysaccharides is the basis for recommending universal vaccination with group A meningococcal polysaccharide twice in infancy, followed by the four-valent vaccine in children aged two and six years. This could eliminate epidemic and endemic disease, prepare for the use of conjugates when they become available, and probably could have prevented the recent epidemics of groups A and W135 meningitis in Burkina Faso. PMID- 14758437 TI - A pioneer in the quest to eradicate world blindness. PMID- 14758438 TI - Intra-ocular acrylic lenses after cataract extraction. 1952. PMID- 14758439 TI - Smallpox and bioterrorism. AB - Smallpox was declared to be eradicated on 8 May 1980, during the Thirty-third World Health Assembly. However, concerns about the possible use of the virus as a weapon of bioterrorism have increased in recent years. Governments have responded by initiating selective vaccination programmes and other public health measures. This review uses historical data from 20th century outbreaks to assess the risks to current populations (which have declining immunity) from a deliberate release of virus. The data presented supports the conclusion of a previous reviewer (Mack) that "smallpox cannot be said to live up to its reputation. Far from being a quick-footed menace, it has appeared as a plodding nuisance with more bark than bite." Its R value (the average number of secondary cases infected by a primary case) is lower than that for measles, human parvovirus, chickenpox, mumps, rubella, and poliomyelitis; only the value for severe acute respiratory syndrome (SARS) is lower. Like SARS, close person-to-person contact is required for effective spread of the disease, and exposure to the virus in hospitals has played an important role in transmission for both viruses. In the present paper the dangers of mass vaccination are emphasized, along with the importance of case isolation, contact tracing, and quarantine of close contacts for outbreak control. The need for rapid diagnosis and the continued importance of maintaining a network of electron microscopes for this purpose are also highlighted. PMID- 14758440 TI - Potential impact of pharmaceuticals on environmental health. PMID- 14758441 TI - France caught cold by heatwave. PMID- 14758442 TI - Drug prices may be too high despite WTO deal. PMID- 14758443 TI - Long-lasting bednets to be produced in Africa. PMID- 14758444 TI - WHO declares failure to deliver AIDS medicines a global health emergency. PMID- 14758445 TI - New low-cost meningitis vaccine developed in record time. PMID- 14758446 TI - Vitamin D deficiency in early life accelerates Type 1 diabetes in non-obese diabetic mice. AB - AIMS/HYPOTHESIS: 1,25-dihydroxyvitamin D(3), the active form of vitamin D, prevents Type 1 diabetes in non-obese diabetic (NOD) mice. Epidemiological data show a threefold increase in human Type 1 diabetes when vitamin D deficiency was present in the first months of life. To evaluate whether a similar dietary deficiency affects diabetes incidence in NOD mice, we generated NOD mice with vitamin D deficiency in early life. METHODS: Breeding pairs of NOD mice, as well as their offspring (test mice), were kept in surroundings devoid of ultraviolet light and were fed a vitamin D-depleted diet for 100 days. Mice were followed for 250 days. RESULTS: At 250 days, 35% (12/35) male and 66% (22/33) female vitamin D deficient mice were diabetic compared to 15% (6/40, p=0.05) and 45% (13/29, p<0.01) of the control mice. At 100 days no difference in insulitis was seen, but more vitamin D-deficient mice were glucose intolerant. Higher IL1 expression was detected in islets of vitamin D-deficient mice and their peritoneal macrophages had an aberrant cytokine profile (low IL1 and IL6, high IL15). Thymus and lymph nodes of vitamin D-deficient mice contained less CD4(+)CD62L(+) cells. CONCLUSION/INTERPRETATION: Vitamin D status increases the expression of Type 1 diabetes in NOD mice. Our data in NOD mice, as well as human epidemiological data, point to the importance of preventing vitamin D deficiency in early childhood. Controlling this dietary factor could be an easy and safe way to reduce the incidence of Type 1 diabetes in subjects who are genetically at risk. PMID- 14758447 TI - Growth of the aorta in children with Williams syndrome: does surgery make a difference? AB - Supravalve aortic stenosis (SAS) and arch hypoplasia are features of Williams syndrome. The effect of aortoplasty on growth of the aorta is not established. We hypothesize that growth of the aorta remains deficient whether or not aortoplasty has been performed. Review of the Children's Hospital of Pittsburgh database revealed 18 patients with Williams syndrome and SAS. Fourteen had sufficient data for inclusion. Patients were divided into two groups based on whether or not they had undergone aortoplasty (groups 1 and 2, respectively). Echocardiographic velocity estimates of the aorta were made at two time points in all patients and one additional time point postoperatively for group 1. Measurements were converted to zeta scores and compared. Peak pulsed echo Doppler velocity (m/sec) in the ascending aorta was higher in patients who underwent aortoplasty. This decreased significantly after surgery. Preoperatively, there was no difference between the groups' annulus, ascending aorta, transverse aorta, and isthmus measurements. At a mean of 43 months postoperatively, there was no significant change in size of the ascending aorta, transverse aorta, and isthmus. Children with Williams syndrome have hypoplasia of the aortic arch that remains constant. Aortoplasty decreases the aortic gradient but has no effect on the size of the ascending aorta, transverse aorta, and aortic isthmus over the short-term. PMID- 14758448 TI - Kidney stone inhibitors in patients with renal stones and endemic renal tubular acidosis in northeast Thailand. AB - Distal renal tubular acidosis (dRTA) is generally associated with hypercalciuria, hypocitraturia, and nephrolithiasis. Our intention was to study glycosaminoglycans (GAGS) and nephrocalcin (NC), two well-known crystal growth inhibitors, in a population with endemic dRTA and nephrolithiasis in northeast (NE) Thailand. We studied 13 patients, six with dRTA and seven with nephrolithiasis with normal or undefined acidification function. Six healthy adults living in the same area as the patients and another six from the Bangkok (BKK) area were used as controls. We measured urinary pH, ammonia, calcium, citrate, magnesium, oxalate, potassium, sodium and uric acid. GAGS were determined by an Alcian blue precipitation method and were qualitated by agarose gel electrophoresis after being isolated using 5% cetyltrimethylammonium bromide at pH 6.0. NC isoforms were isolated as previously described by Nakagawa et al. Citrate was higher in BKK controls ( p<0.04). There was a striking difference among GAGS from BKK when compared with other groups (103.85+/-10.70 vs. 23.52+/ 8.11 for dRTA, 22.36+/-14.98 for kidney stone patients and 14.73+/-2.87 mg/ml in controls from the NE region, ( p<0.0001). dRTA and stone-forming patients excrete proportionally more (C+D) than (A+B) NC isoforms ( p<0.05). Also, their NC showed a 100-fold weaker binding capacity of calcium oxalate monohydrate crystals. The ratio of chondroitin sulfate/heparin sulfate in GAGS was approximately 9/1. In addition to the traditional risk factors for nephrolithiasis in dRTA, GAGS and NC might play an important role in the pathogenesis of stone formation in this population. PMID- 14758450 TI - PHACES syndrome: a review of eight previously unreported cases with late arterial occlusions. AB - PHACE and PHACES are acronyms for a syndrome of variable expression comprising posterior cranial fossa malformations, facial haemangiomas, arterial anomalies, aortic coarctation and other cardiac disorders, ocular abnormalities and stenotic arterial disease. We review five girls and three boys aged 1 month-14 years with disorders from this spectrum. Six had large facial haemangiomas but recent reports suggest that small haemangiomas may occur; hence our inclusion of two possible cases. We also focus on the recently recognised feature of progressive intracranial arterial occlusions, present in four of our patients, later than previously recognised, from 4 to 14 years of age. We suggest that many elements of this disorder could reflect an abnormality of cell proliferation and apoptosis. PMID- 14758451 TI - Embolisation of both fistulae through the same carotid artery tear in a patient with bilateral traumatic caroticocavernous fistulae. AB - Endovascular treatment of traumatic caroticocavernous fistulae (CCF) may present technical difficulties with specific angiographic dilemmas. We report endovascular techniques used in a patient with bilateral post-traumatic CCF, high flow on one side, and slow-flow on the other. Complete closure of both was achieved through the same carotid artery tear. To our knowledge, transarterial venous coil embolisation of a low-flow fistula through a contralateral carotid artery tear, with transarterial detachable balloon embolisation of the ipsilateral high-flow fistula has not been described previously. PMID- 14758452 TI - Cerebellar damage produces context-dependent deficits in control of leg dynamics during obstacle avoidance. AB - It has been suggested that the cerebellum is an important contributor to CNS prediction and control of intersegmental dynamics during voluntary multijoint reaching movements. Leg movements subserve different behavioral goals, e.g., locomotion versus voluntary stepping, which may or may not be under similar dynamic control. The objective was to determine whether cerebellar leg hypermetria (excessive foot elevation) during obstacle avoidance in locomotion and voluntary stepping could be attributed to a particular deficit in appropriately controlling intersegmental dynamics. We compared the performance of eight individuals with cerebellar damage to eight healthy controls as they walked or voluntarily stepped in place over a small obstacle. Joint kinematics and dynamics were calculated during swing phase for both movement contexts. The kinematic analysis showed that hypermetria occurred during both walking and stepping and was associated with excessive knee flexion. When present, the amplitude of hypermetria was greater during stepping compared to walking. During stepping, subjects with cerebellar damage produced excessive knee flexor muscle torques and consequently overcompensated for interaction and gravitational torques normally used to decelerate the limb. During walking, the torque pattern was very similar to that of control subjects walking over a taller obstacle, and therefore might be a voluntary compensatory strategy to avoid tripping. Our results show that the extent of kinematic and dynamic abnormalities associated with cerebellar leg hypermetria is context-specific, with more fundamental abnormalities of leg dynamics being apparent during stepping as opposed to walking. PMID- 14758454 TI - Effects of bilateral vestibular loss on podokinetic after-rotation. AB - We asked what the role of the vestibular system is in adaptive control of locomotor trajectory in response to walking on a rotating disc. Subjects with bilateral vestibular loss (BVL) were compared to age- and gender-matched controls (CTRL). Subjects walked in place on the surface of a rotating disc for 15 min and then attempted to step in place without vision on a stationary surface for 30 min. CTRL subjects demonstrated podokinetic after-rotation (PKAR), involuntarily and unknowingly turning themselves in circles while attempting to step in place. PKAR in CTRLs was characterized by a rapid rise in turning velocity over the first 1-2 min, followed by a gradual decay over the remaining 28 min. Subjects with BVL also demonstrated PKAR and had no knowledge of their turning. However, PKAR in BVLs was characterized by an extremely rapid, essentially instantaneous rise. Subjects with BVL immediately turned at maximum velocity and exhibited a gradual decay throughout the entire 30 min period. Despite this difference in the initial portion of PKAR in BVLs, their responses were not significantly different from CTRLs during minutes 2 to 30 of the response. These results suggest that vestibular inputs normally suppress PKAR velocity over the first 1-2 min of the response, but do not greatly influence PKAR decay. PKAR is therefore a process mediated primarily by somatosensory information and vestibular inputs are not required for its expression. Additionally, the absence of vestibular inputs does not result in increased somatosensory sensitivity that alters podokinetic intensity or decay time constants. PMID- 14758453 TI - Resolving conflicts in task demands during balance recovery: does holding an object inhibit compensatory grasping? AB - The ability to reach and "grasp" (grip or touch) structures for support in reaction to instability is an important element of the postural repertoire. It is unclear, however, how the central nervous system (CNS) resolves the potential conflict between holding an object and the need to release the held object and grasp alternative support, particularly if the held object is perceived to be relevant to the task of stabilizing the body, e.g. an assistive device. This study examined whether compensatory grasping is inhibited when holding an object, and whether the influence differs when holding an assistive device (cane) versus a task-irrelevant object (top handle portion of a cane). We also investigated the influence of preloading the assistive device, to determine whether conflicting demands for arm-muscle activation (requiring disengagement of ongoing agonist or antagonist activity) would influence the inhibition of compensatory grasping. Unpredictable forward and backward platform translations were used to evoke the balancing reactions in 16 healthy young adults. A handrail was mounted to the right and foot motion was constrained by barriers, with the intent that successful balance recovery would (in large-perturbation trials) require subjects to release the held object and contact the rail with the right hand. Results showed that grasping reactions were commonly used to recover equilibrium when the hand was free (rail contact in 71% of large-perturbation trials). However, holding either the cane or canetop had a potent modulating effect: although early biceps activation was almost never inhibited completely (significant activity within 200 ms in 98% of trials), the average activation amplitude was attenuated by 30-64% and the average frequency of handrail contact was reduced by a factor of two or more. This reduced use of the rail occurred even though the consequence often involved falling against a safety harness or barriers. Handrail contact occurred least frequently when holding the cane during forward loss of balance: subjects persisted in pushing on the cane (failing to use the rail) in 93% of trials, even when the perturbations were too large to allow this strategy to be successful. Prior contraction (preloading the cane) did not influence any of these findings. Complex strategies (e.g. partial release of object) were often adopted to allow balance to be recovered without dropping the held object. Remarkably, it appears that the CNS may give priority to the ongoing task of holding an object, even when it has no stabilizing value (cane during backward falls) or any intrinsic value whatsoever (canetop). PMID- 14758457 TI - Lab-on-a-chip systems for biomedical and environmental monitoring. AB - During the last decade, pocket-sized analytical equipment based on the "lab-on-a chip" approach has become available. These chips, in combination with portable electronic equipment, are applicable in, for example, "point-of-care" ion analysis of body fluids, forensics, identification of explosives, tracking of pollution in environmental or waste waters, monitoring nutrients in agricultural or horticultural water, controlling quality in food production, or process control in chemical industry. This paper discusses several demonstrator systems with applications in these fields. PMID- 14758460 TI - Peptide recognition via hierarchical imprinting. AB - Silica particles containing immobilised peptidic templates have been used for the generation of hierarchically imprinted polymers. The pores of the silica mould were filled with a mixture of monomers/initiator and polymerised, followed by dissolution of the silica template. This method leaves behind imprinted polymers with binding sites located at the surface, which are capable of recognising larger molecules with the same immobilised epitope. All the products resulting from solid-phase synthesis of peptides were characterised by elemental analysis, FT-IR spectroscopy and fluorescence microscopy. The hierarchically imprinted polymers generated from these products were characterised by elemental analysis, FT-IR spectroscopy, fluorescence microscopy, scanning electron microscopy (SEM) and nitrogen adsorption, providing evidence concerning the reproducibility of each step. The chromatographic properties of the materials have been investigated and the advantages of the immobilisation method have been proven. The materials exhibit selectivity for their templates and other structurally related dipeptides. Furthermore, the polymers proved to be capable of recognising larger peptides containing the immobilised sequence. PMID- 14758462 TI - Sensitive biomonitoring of phthalate metabolites in human urine using packed capillary column switching liquid chromatography coupled to electrospray ionization ion-trap mass spectrometry. AB - A rapid and sensitive method for the determination of the phthalate monoesters monoethyl phthalate (MEP), monobutyl phthalate (MBP), monobenzyl phthalate (MBzP) and monoethylhexyl phthalate (MEHP), in human urine, using packed capillary column liquid chromatography coupled to electrospray quadrupole-ion trap mass spectrometry (ESI-QITMS(n)) has been developed. Sample volumes of 200 microL of deconjugated and diluted urine were loaded onto a precolumn of 30 mmx0.32 mm I.D. packed with Hypercarb 5 microm particles, using a sample carrier consisting of acetonitrile/water (15/85, v/v, adjusted to pH 2 using HCl) with a flow rate of 20 microL/min. Backflushed elution onto a 100 mmx0.32 mm I.D. analytical column packed with 5 microm Hypercarb particles was conducted using a tetrahydrofuran/water gradient where both solvents contained 10 mM ammonium acetate, at a flow rate of 4 microL/min. Determination of the monophthalates was achieved within 8 min. Ionization was performed in the negative mode and the analytes were observed as [M-H](-) at m/ z=193.1, 221.1, 255.1 and 277.0 for MEP, MBP, MBzP and MEHP, respectively. Quantification was performed in the multiple reaction monitoring (MRM) mode monitoring the fragments at m/ z=121.1, 177.0, 183.0 and 233.0 for MEP, MBP, MBzP and MEHP, respectively. The method was validated over the concentration range 2.5-125 ng/mL in pretreated urine samples, corresponding to 25-1250 ng/mL untreated urine, yielding correlation coefficients in the range 0.996-0.999. The within-assay ( n=6) and between-assay ( n=6) repeatabilities were in the range 4.0-18% and 4.8-15% RSD, respectively. The mass limits of detection were in the range 32-70 pg, corresponding to concentration limits of detection of 1.6-3.5 ng/mL of untreated urine. PMID- 14758463 TI - Analysis of bulk and inorganic degradation products of stones, mortars and wall paintings by portable Raman microprobe spectroscopy. AB - This work reports the use of a portable Raman microprobe spectrometer for the analysis of bulk and decaying compounds in carbonaceous materials such as stones, mortars and wall paintings. The analysed stones include limestone, dolomite and carbonaceous sandstone, gypsum and calcium oxalate, both mono- and dihydrated, being the main inorganic degradation products detected. Mortars include bulk phases with pure gypsum, calcite and mixtures of both or with sand, soluble salts being the most important degradation products. The pigments detected in several wall paintings include Prussian blue, iron oxide red, iron oxide yellow, vermilion, carbon black and lead white. Three different decaying processes have been characterised in the mortars of the wall paintings: (a) a massive absorption of nitrates that reacted with calcium carbonate and promoted the unbinding of pigment grains, (b) the formation of black crusts in the vault of the presbytery and (c) the thermodecomposition of pigments due to a fire. PMID- 14758464 TI - Determination of ochratoxin A in pig tissues by liquid-liquid extraction and clean-up and high-performance liquid chromatography. AB - A fast, simple, and sensitive HPLC-FD method is described for determination of ochratoxin A (OTA) in pig kidney and muscle; a small mass (<2.5 g) of sample and a relatively small volume (<15 mL) of a non-halogenated extraction solvent are required. Ochratoxin B, systematically absent from all the samples investigated, was used as internal standard. Liquid-liquid partition was used for sample clean up. Recoveries at the 1 ng x g(-1) level were 86+/-15% and 74+/-8% for kidney and muscle, respectively, and detection limits were 0.14 and 0.15 ng x g(-1). Clean up by solid-phase extraction (SPE) is required for pig liver. A survey of the OTA content of tissues of pigs slaughtered in southern Italy revealed that 52 out of 54 analysed samples were contaminated; the OTA concentration in kidney ranged between 0.26 and 3.05 ng x g(-1). The effect of measurement precision on compliance with legal limits is also discussed. PMID- 14758465 TI - Determination of polychlorobiphenyls and polycyclic aromatic hydrocarbons in the atmospheric aerosol of the Venice Lagoon. AB - An analytical method for simultaneous determination of "particle"-associated and "gaseous"-phase concentrations of polychlorinated biphenyls (PCB) and polycyclic aromatic hydrocarbons (PAH) in atmospheric aerosol samples obtained by high volume samplers using polyurethane foam adsorbent (PUF) and quartz fibber filters (QFF) has been investigated. Quality control of the analytical procedure was carried out by blank control and by evaluating limits of detection, recoveries, accuracy, and repeatability. The proposed method was subsequently used to determine PAH and PCB in the "gaseous" and "particulate" phases of the aerosols that enter the Venice Lagoon atmosphere. The highest concentrations of sigma PCB and sigma PAH were predominantly in the "gaseous" phase. In both "particulate" and "gaseous" phases the penta-CB congeners dominated total PCB concentrations whereas phenanthrene, fluoranthene, and pyrene dominated the sigma PAH concentrations. Total ("gaseous" plus "particulate") PCB and PAH concentrations were higher at the site directly influenced by the industrial plants but the concentrations in marine aerosol samples were lower by a factor four only and must be taken into consideration when studying the chemical contamination of the Venice Lagoon. PMID- 14758466 TI - Monitoring ground-level air for trace analysis: methods and results. AB - Trace analysis enables the sensitive detection of radionuclide concentrations in ground-level air in the range of microbecquerel per cubic meter (microBq m(-3)). Typical sampling intervals of less than one day up to a few days can be used in routine operation. Trace analysis measurements are performed in the framework of the German Integrated Measuring and Information system (IMIS) and the International Monitoring System (IMS) used for verification of the Comprehensive Nuclear-Test-Ban Treaty (CTBT). Within the environmental monitoring programmes of the German IMIS the Federal Office for Radiation Protection (BfS) performs measurements of aerosol-bound radionuclides and of radioactive noble gases in the atmosphere. Aerosols are collected on filters with high-volume air samplers and analysed by gamma-spectrometry, alpha-spectrometry, and integral measurements of beta-activity, with preceding radiochemical separation. Noble gas samples from 15 sites world-wide are analysed to observe the (85)Kr-release from nuclear fuel reprocessing plants and from seven sites in Germany to monitor the (133)Xe emitted from nuclear power plants. As part of the International Monitoring System (IMS) of the CTBT an automatic aerosol sampling and measuring system and an automatic noble gas sampling and measuring system will be operated by the BfS at Mount Schauinsland near Freiburg. Because of its expertise in noble gas measurements the BfS had been chosen to perform an intercomparison experiment in the BfS laboratory in Freiburg with several automatic noble gas sampling and measurement systems before their installation at IMS sites. To establish quality assurance programmes for trace analysis performed for the German IMIS close collaboration between the involved German institutions has been established. First steps have been taken to expand cooperation to other European laboratories. Informal data exchange already occurs between trace-analysis laboratories in Europe (Ring of Five) and helps in cases of enhanced activity concentrations to get a rapid overview of the radiological situation and to identify possible sources. PMID- 14758467 TI - Study of the behavioural responses related to the potential addictive properties of MDMA in mice. AB - We investigated several behavioural responses induced by repeated administration of MDMA in mice that could be related to its potential abuse liability. Mice treated with MDMA at the dose of 10 mg/kg displayed a significant conditioned place preference with respect to saline treated controls, while lower doses (0.3, 1.0, 3.3 mg/kg) had no effect. The development of physical dependence was also investigated. Mice were treated with MDMA (10 mg/kg) twice daily for 5 days. On day 6, following a single administration of MDMA mice received the following monoaminergic antagonists: metergoline (0.1 and 1 mg/kg), ritanserin (0.25 and 1 mg/kg), timolol (2 and 10 mg/kg), prazosin (0.25 and 1 mg/kg), SCH 23390 (0.05 and 0.25 mg/kg), raclopride (0.1 and 0.5 mg/kg) or vehicle, and several somatic manifestations of withdrawal were evaluated for 45 min. Metergoline induced paw tremor, face rubbing, as well as an increase in locomotor activity in mice chronically treated with MDMA. Ritanserin, and timolol induced only paw tremor, while SCH 23390 and raclopride did not produce any somatic manifestation indicative of abstinence. The possible modification of the rewarding properties of MDMA (10 mg/kg) by the monoaminergic antagonists producing the most relevant somatic signs of withdrawal namely, metergoline (0.1 and 1 mg/kg) and timolol (2 and 10 mg/kg) were tested in the conditioned place preference paradigm. Results showed that metergoline did not significantly modify the rewarding properties of MDMA, whereas only the highest dose of timolol was able to decrease MDMA reward. No signs of dopaminergic neurotoxicity were observed following chronic treatment with MDMA as revealed by [(3)H] mazindol binding. The possible motivational and affective components of the withdrawal syndrome were assessed in the suppression of operant responding for food, the conditioned place aversion, and the lit/dark paradigms. Results showed that the somatic symptoms observed were not accompanied by any aversive/dysphoric or anxiogenic-like behaviours. These results reveal the rewarding properties of MDMA in mice, and suggest that chronic MDMA administration does not induce classical manifestations of physical dependence in mice. PMID- 14758468 TI - Pharmacology of a novel selective 5-hydroxytryptamine1B receptor antagonist, AR A000002. AB - The terminal 5-HT(1B) autoreceptors have attracted great pharmacological interest since they are potential targets for compounds modifying serotonergic neurotransmission. In the present work the in vivo biochemical properties of AR A000002 ((R)-N-[5-methyl-8-(4-methylpiperazin-1-yl)-1,2,3,4-tetrahydro-2 naphthyl]-4-morpholinobenzamide), a novel selective 5-HT(1B) receptor antagonist, are reported. The effects of AR-A000002 on: 5-HT metabolism was measured as the ratio between 5-HIAA and 5-HT concentrations in different brain regions; 5-HT synthesis was measured as the accumulation of 5-HTP after inhibition of the aromatic amino acid decarboxylase activity with m-hydroxybenzylhydrazine (NSD1015); 5-HT release was measured using the microdialysis technique. 5-HT, 5 HIAA and 5-HTP concentrations were analyzed using high power liquid chromatography (HPLC) with electrochemical detection. AR-A000002 significantly enhanced 5-HT metabolism (5-HIAA/5-HT ratio) and 5-HT synthesis in guinea pig brain in the dose range 0.9-18 mg/kg s.c. (ED(50)=1 mg/kg s.c. in the four brain regions examined) with maximal effect seen after 2-4 h. AR-A000002 (9 mg/kg s.c.) significantly increased the extracellular concentrations of 5-HT and 5-HIAA by 20% in the guinea pig frontal cortex, measured with the in vivo microdialysis technique in freely moving guinea pigs. AR-A000002 (9 mg/kg s.c.) in combination with the 5-HT uptake inhibitor citalopram (5 mg/kg s.c.) increased the extracellular 5-HT concentration in guinea pig frontal cortex from 250 to 400% of the basal level. Citalopram alone decreased the extracellular 5-HIAA levels to 70% of the basal value. AR-A000002 counteracted the citalopram-induced decrease in 5-HIAA. Since the basal level of extracellular 5-HIAA was 160 times higher than that of 5-HT the 20% increase in 5-HIAA concentrations indicates that only a few percent of the exocytotically released 5-HT from the nerve terminals reached the extracellular space when the re-uptake mechanism was intact. The results also show that the terminal 5-HT(1B) autoreceptors are tonically activated under drug free as well as citalopram conditions. The increase in plasma level of cortisol after AR-A000002 administration may indicate stimulation of post-synaptic 5-HT receptors. AR-A000002 also blocked 5-HT(1B) agonist-induced (CP-135,807) decrease in 5-HT metabolism and hypothermia (ED(50)=1 mg/kg s.c.), thus indicating competition between these two drugs. It is concluded that AR-A000002 is a 5 HT(1B) receptor antagonist that enhances the serotonergic neurotransmission in guinea pig brain. PMID- 14758469 TI - Phenyl methyl ethers: novel electron donors for respiratory growth of Desulfitobacterium hafniense and Desulfitobacterium sp. strain PCE-S. AB - Desulfitobacterium hafniense and Desulfitobacterium sp. strain PCE-S grew under anoxic conditions with a variety of phenyl methyl ethers as electron donors in combination with fumarate as electron acceptor. The phenyl methyl ethers were O demethylated to the corresponding phenol compounds. O-demethylation was strictly dependent on the presence of fumarate; no O-demethylation occurred with CO2 as electron acceptor. One mol phenyl methyl ether R-O-CH3 was O-demethylated to R-OH per 3 mol fumarate reduced to succinate. The growth yields with vanillate or syringate plus fumarate were approximately 15 g cells (dry weight) per mol methyl moiety converted. D. hafniense utilized vanillate or syringate as an electron donor for reductive dehalogenation of 3-Cl-4-hydroxyphenylacetate, whereas strain PCE-S was not able to dechlorinate tetrachloroethene with phenyl methyl ethers. Crude extracts of both organisms showed O-demethylase activity in the O demethylase assay with vanillate or syringate as substrates when the organism was grown on syringate plus fumarate. Besides the homoacetogenic bacteria, only growing cells of Desulfitobacterium frappieri PCP-1 have thus far been reported to be capable of phenyl methyl ether O-demethylation. This present study is the first report of Desulfitobacteria utilizing phenyl methyl ethers as electron donors for fumarate reduction and for growth. PMID- 14758470 TI - Cloning, identification and expression of an entE homologue angE from Vibrio anguillarum serotype O1. AB - Anguibactin, an important virulent factor in Vibrio anguillarum serotype O1, is synthesized by a nonribosomal peptide synthetases (NRPS) system encoded on a 65 kb virulence plasmid pJM1. angE, as one of the NRPS genes, is responsible for selecting and activating 2,3-dihydroxybenzoic acid (2,3-DHBA), an important precursor in anguibactin synthesis, into 2,3-DHBA-AMP by adenylylation in the presence of ATP. In this work, an entE homologue, angE, was identified on pEIB1 (a pJM1-like plasmid) from virulent V. anguillarum serotype O1 strain MVM425. A recombinant clone carrying the complete angE was able to complement an Escherichia coli entE mutant. The angE-encoded protein was overexpressed in E. coli and purified by a three-step procedure. Purified AngE was then used to establish an in vitro enzymatic reaction in which its enzymatic activity of 1-(5' monophosphate adenyl) 2,3-dihydroxybenzoic acid ligase (2,3-DHBA-AMP ligase) was proved using HPLC to detect AMP formation in the reaction mixture. Moreover, evidence at the level of both transcription and translation confirmed that angE was actively expressed in vivo in V. anguillarum MVM425, and interestingly, unlike many other iron-uptake-system-related genes, its expression is not induced by a low iron concentration in the surrounding environment. PMID- 14758471 TI - Silicone rubber: an alternative for repair of articular cartilage defects. AB - Silicone-rubber implants were used to fill full-thickness articular cartilage in the trochlea area of the knee joint in rabbits, for the purpose of studying the long-term influence of silicone-rubber implant on surrounding articular cartilage. Forty eight weeks after surgery, the silicone rubbers were still fitted tightly into the defects; surrounding cartilage showed mild degeneration, better than the control group. Our results showed silicone-rubber implantation for repairing local articular cartilage defects can effectively delay the pathogenetic progression of osteoarthritis. PMID- 14758472 TI - Prevalence of intra-abdominal hypertension in critically ill patients: a multicentre epidemiological study. AB - OBJECTIVE: Although intra-abdominal hypertension (IAH) can cause dysfunction of several organs and raise mortality, little information is available on the incidence and risk factors for IAH in critically ill patients. This study assessed the prevalence of IAH and its risk factors in a mixed population of intensive care patients. DESIGN: A multicentre, prospective 1-day point prevalence epidemiological study conducted in 13 ICUs of six countries. INTERVENTIONS: None. PATIENTS: Ninety-seven patients admitted for more than 24 h to one of the ICUs during the 1-day study period. METHODS: Intra-abdominal pressure (IAP) was measured four times (every 6 h) by the bladder pressure method. Data included the demographics, medical or surgical type of admission, SOFA score, etiological factors such as abdominal surgery, haemoperitoneum, abdominal infection, massive fluid resuscitation, and ileus and predisposing conditions such as hypothermia, acidosis, polytransfusion, coagulopathy, sepsis, liver dysfunction, pneumonia and bacteraemia. RESULTS: We enrolled 97 patients, mean age 64+/-15 years, 57 (59%) medical and 40 (41%) surgical admission, SOFA score of 6.5+/-4.0. Mean IAP was 9.8+/-4.7 mmHg. The prevalence of IAH (defined as IAP 12 mmHg or more) was 50.5 and 8.2% had abdominal compartment syndrome (defined as IAP 20 mmHg or more). The only risk factor significantly associated with IAH was the body mass index, while massive fluid resuscitation, renal and coagulation impairment were at limit of significance. CONCLUSION: Although we found a quite high prevalence of IAH, no risk factors were reliably associated with IAH; consequently, to get valid information about IAH, IAP needs to be measured. PMID- 14758473 TI - Action induction through action observation. AB - Ideomotor movements may arise in individuals while they watch goal-directed actions or events. In a previous study we developed a paradigm for investigating ideomotor movements induced through watching the outcome of one's own action. In the present study we extended the paradigm to investigate both movements induced through watching the outcome of one's own as well as somebody else's action (player mode and observer mode respectively). We report three experiments, each with differing conditions for the player mode, but identical conditions for the observer mode. Results indicate that in both modes ideomotor movements are governed by two basic principles: Perceptual induction and intentional induction. In the player mode we replicated and extended previous findings, indicating dissociation between hand and head movements. In the observer mode no such dissociation was obtained. Our findings suggest that people perform, in their own actions, what they see being performed in other people's actions. Induction of action through observation can pertain to both the action's physical surface and underlying intentions. Furthermore, our results suggest that perceptual induction is ubiquitous but may be locally suspended for intentional action control. We discuss our results in the framework of theories invoking a strong overlap between representational structures for action perception and action planning. PMID- 14758474 TI - Aspects of temporal information processing: a dimensional analysis. AB - A major controversy in the field of prospective temporal information processing refers to the question of whether performance in various temporal tasks can be accounted for by the general assumption of an internal clock rather than by distinct, task-specific timing mechanisms. Therefore, the present study was designed to identify dimensions of temporal information processing. For this purpose, 120 subjects performed eight psychophysical temporal tasks. Correlational and principal factor analyses suggested a common pacemaker-based interval timing mechanism involved in duration discrimination, temporal generalization, and temporal order judgment. On the other hand, rhythm perception and perceived simultaneity/successiveness appeared to be controlled by task specific processes unrelated to interval-based timing. PMID- 14758476 TI - Novel eceriferum mutants in Arabidopsis thaliana. AB - We conducted a novel non-visual screen for cuticular wax mutants in Arabidopsis thaliana (L.) Heynh. Using gas chromatography we screened over 1,200 ethyl methane sulfonate (EMS)-mutagenized lines for alterations in the major A. thaliana wild-type stem cuticular chemicals. Five lines showed distinct differences from the wild type and were further analyzed by gas chromatography and scanning electron microscopy. The five mutants were mapped to specific chromosome locations and tested for allelism with other wax mutant loci mapping to the same region. Toward this end, the mapping of the cuticular wax ( cer) mutants cer10 to cer20 was conducted to allow more efficient allelism tests with newly identified lines. From these five lines, we have identified three mutants defining novel genes that have been designated CER22, CER23, and CER24. Detailed stem and leaf chemistry has allowed us to place these novel mutants in specific steps of the cuticular wax biosynthetic pathway and to make hypotheses about the function of their gene products. PMID- 14758475 TI - Immunocytochemical localization of polygalacturonase during tracheary element differentiation in Zinnia elegans. AB - Polygalacturonase (PG) is a cell wall-associated protein that degrades pectin. A ZePG1 cDNA encoding a putative PG was isolated from Zinnia elegens L. and a rabbit antibody specific to the ZePG1 protein was generated. The level of the ZePG1 protein was up-regulated when tracheary element differentiation was initiated. Using gold-labeled secondary antibodies for light and electron microscopy, ZePG1 protein was localized in cultured Zinnia cells. This protein was preferentially distributed on tracheary elements (TEs). At the subcellular level, the protein was localized on secondary wall thickenings, primary walls, Golgi bodies and vesicles. Thus, the putative role of the ZePG1 protein might be the degradation of pectic substances before lignification. Some non-TE cells also accumulated ZePG1 protein on primary walls, Golgi bodies and vesicles. The accumulation of ZePG1 protein on primary walls seems to be at the elongating tips of non-TE cells. In plants, ZePG1 protein was localized on the secondary wall thickenings of differentiating TEs and phloem regions. These results suggest that the expression of the ZePG1 protein is highly regulated both spatially and temporally during in vitro and in situ TE differentiation. PMID- 14758477 TI - The slow component of oxygen uptake during intense, sub-maximal exercise in man is associated with additional fibre recruitment. AB - Single muscle fibre metabolites and pulmonary oxygen uptake (VO2) were measured during moderate and intense, sub-maximal exercise to test the hypothesis that additional fibre recruitment is associated with the slow component of VO2. Seven healthy, male subjects performed 20 min moderate (MOD, approximately 50% of VO(2,max)) and intense (INT, approximately 80% VO(2,max)) cycling at 70 rpm. Glycogen content decreased significantly in type I and IIa fibres during INT, but only in type I fibres during MOD. During INT, creatine phosphate (CP) content decreased significantly both in types I and II fibres in the first 3 min (DeltaCP: 16.0+/-2.7 and 16.8+/-4.7 mmol kg(-1) d.w., respectively) and in the next 3 min (DeltaCP: 16.2+/-4.9 and 25.7+/-6.7 mmol kg(-1) d.w., respectively) with no further change from 6-20 min. CP content was below the pre-exercise level (mean-1 SD) in 11, 37, 70 and 74% of the type I fibres after 0, 3, 6 and 20 min of INT, respectively, and in 13, 45, 83 and 74% of the type II fibres. During INT, VO2 increased significantly by 6+/-1 and 4+/-1% in the periods 3-6 and 6-20 min, respectively (Delta VO(2,(6-3 min)): 0.14+/-0.02 l min(-1)), whereas VO2 was unchanged from 3 to 20 min of MOD. Exponential fitting revealed a slow component of VO2 during INT that appeared after approximately 2.6 min and amounted to 0.24 l min(-1). The present study demonstrates that additional type I and II fibres are recruited with time during intense sub-maximal exercise in temporal association with a significant slow component of VO2. PMID- 14758478 TI - Intracellular nucleotides and polyamines inhibit the Ca2+-activated cation channel TRPM4b. AB - TRPM4b (in contrast to the short splice variant TRPM4a) is a Ca(2+)-activated but Ca(2+)-impermeable cation channel. We have studied TRPM4 currents in inside-out patches. Supramicromolar Ca(2+) concentrations applied at the inner side, [Ca(2+)](i), activated TRPM4 with an EC(50) value of 0.37 mM, a value that is much higher than that of whole-cell currents. Current amplitudes decreased above 1 mM [Ca(2+)](i), (IC(50) 9.3 mM). Sr(2+) but not Ba(2+)could partially substitute for Ca(2+). ATP, ADP, AMP and AMP-PNP all quickly and reversibly inhibited TRPM4 with IC(50) values between 2 and 19 microM (at +100 mV). Adenosine also blocked TRPM4 at 630 microM. The block at high ATP concentrations was incomplete and was not affected by the presence of free Mg(2+). ADP induced the most sensitive block with an IC(50) of 2.2 microM. For inhibition of TRPM4 by free ATP(4-), an IC(50) value of 1.7+/-0.3 microM was calculated. GTP, UTP and CTP at concentrations up to 1 mM did not induce a similar block. Spermine blocked TRPM4 currents with an IC(50) of 61 microM. In conclusion, TRPM4 is a channel that can be effectively modulated by intracellular nucleotides and polyamines. PMID- 14758479 TI - Calcineurin and heat-shock proteins modulation in clenbuterol-induced hypertrophied rat skeletal muscles. AB - To examine the changes in heat shock proteins (HSPs) and calcineurin (CaN), a calcium/calmodulin regulated protein phosphatase, in hypertrophied rat skeletal muscles, adult male Wistar rats were administered clenbuterol (CLB, 30 mg l(-1) in drinking water), a beta 2-agonist, for 4 weeks. Compared to controls, CLB treated rats had significantly larger body (10%) and relative (to body weight) soleus (Sol, 16%), plantaris (Plt, 32%) and gastrocnemius (Gast, 27%) weights. Immunohistochemically classified fast fibers were hypertrophied in the Sol (64%), Plt (62%), and deep (d, 70%) and superficial (s, 44%) regions of the Gast, whereas slow fibers were hypertrophied only in the Plt (47%). The percentage of fast fibers in the Sol increased from 10% to 21%. The myosin heavy chain (MHC) isoform composition shifted from slow to fast in the Sol (increase in the percentage of type IIa MHC and de novo synthesis of type IIx MHC) and Gast-d (de novo synthesis of type IIb MHC) and to the faster isoforms in the Plt (increase in the percentage of type IIb MHC). Hsp72 and Hsp90 levels in CLB-treated rats were 52% and 50% lower in the Sol and 44 and 41% lower in the Gast-d, respectively, than in control rats. In control rats, the relative content of CaN was: Sol>Gast-d>Plt>Gast-s, and CLB treatment enhanced the CaN content by 1.4-, 1.2-, 5.0-, and 3.3-fold, respectively. These results indicate that the down regulation of HSPs in the Sol and Gast-d was closely related to a decrease in the slow phenotype, and that the relative up-regulation of CaN among the muscles/regions was closely related to the selective hypertrophy of fast fibers in the CLB-treated rats. PMID- 14758480 TI - Postnatal expression of transport proteins involved in acid-base transport in mouse kidney. AB - The kidney plays a major role in maintaining and controlling systemic acid-base homeostasis by reabsorbing bicarbonate and secreting protons and acid equivalents, respectively. During postnatal kidney development and adaptation to changing diets, plasma bicarbonate levels are increasing, the capacity for urinary acidification maturates, and the final morphology and distribution of intercalated cells is achieved. In adult kidney, at least two types of intercalated cells (IC) are found along the collecting duct characterised either by the expression of AE1 (type A IC) or pendrin (non-type A IC) where non-type A IC are found only in the convoluted distal tubule, connecting tubule and cortical collecting duct. Here we investigated in mouse kidney the relative mRNA abundance, protein expression levels and distribution of several proteins involved in renal acid-base transport, namely, the Na(+)/HCO(3)(-) cotransporter NBC1 (SLC4A4), the Na(+)/H(+)-exchanger NHE3 (SLC9A3), two subunits of the vacuolar H(+)-ATPase [ATP6V0A4 (a4), ATP6V1B1 (B1)], the Cl(-)/HCO(3)(-) exchangers AE1 (SLC4A1) and pendrin (SLC26A4). Relative mRNA abundance of all transport proteins was lowest at day 3 after birth and increased thereafter in parallel with protein levels. The numbers of type A and non-type A IC in the cortical collecting duct (CCD) increased from day 3 to days 18 and 24, whereas the number of IC in the CCD with apical staining for the vacuolar H(+)-ATPase subunits a4 and B1 decreased from day 3 to days 18 and 24, respectively. In addition, cells with characteristics of non-type A IC (pendrin expression, basolateral expression of vacuolar H(+)-ATPase subunits) were found in the inner and outer medulla 3 days after birth but were absent from the medulla of 24-day old mice. Taken together, these results demonstrate massive changes in mRNA and protein expression levels of several acid-base transporters during postnatal kidney maturation and also show changes in intercalated cell phenotype in the medulla during these processes. PMID- 14758481 TI - Walking modality affects respiratory muscle action and contribution to respiratory effort. AB - We hypothesized that walking at increased speed or increasing gradient might have different effects on chest wall kinematics and respiratory muscle power components, and contribute differently to respiratory effort sensation. We measured the volumes of chest wall compartments by optoelectronic plethysmography, esophageal, gastric and transdiaphragmatic ( P(di)) pressures, and the sensation of the respiratory effort by a Borg scale in five normal subjects walking both at ascending gradient with constant speed (AG) and at ascending speed with constant gradient (AS). Chest wall kinematics, evaluated by displacement of chest wall compartments, did not show any significant difference between AS and AG. Muscle power, calculated as the product of mean flow and mean pressure, increased similarly, but its partitioning into pressure and velocity of shortening differed in the two modes. A greater increase in the pressure developed by the abdominal muscles ( P(abm)) (4.06-fold), and in the velocity of shortening of both rib cage inspiratory muscles ( v(rcm,i)) (2.01-fold) and the diaphragm ( v(di)) (1.90-fold) was associated with a lower increase in the pressure developed by the rib cage inspiratory muscles ( P(rcm,i)) (1.24-fold) and P(di) (0.99-fold) with AG. Instead, with AS, a lower increase in P(abm) (2.12 fold), v(rcm,i) (1.66-fold) and v(di) (1.54-fold) was associated with a greater increase in P(rcm,i) (1.56-fold) and P(di) (1.97-fold). A combination of P(abm) and v(di) during AG (Wald chi(2)=23.19, P<0.0000), with the addition of P(rcm,i) during AS (Wald chi(2)=29.46, P<0.0000), was the best predictor of Borg score. In conclusion, the general strategy adopted by respiratory centers during different walking modes does not differ in terms of ventilation, chest wall kinematics, and respiratory muscle power production, whereas it does in terms of partitioning of power into pressure and velocity of shortening, and respiratory muscle contribution to respiratory effort sensation. Combinations of different patterns of flow and pressure generation made the respiratory effort sensation similar during AS and AG modes. PMID- 14758482 TI - Carcinoembryonic antigen (CEA) and CEA-related cell adhesion molecule 1 (CEACAM1), apically expressed on human colonic M cells, are potential receptors for microbial adhesion. AB - In the human gut mucosa, specialized M cells deliver intact foreign macromolecules and commensal bacteria from the lumen to organized mucosal lymphoid tissues triggering immune responses. M cells are also major sites of adhesion and invasion for enteric pathogens. The molecular features of M cell apical surfaces that promote microbial normal attachment are still largely unknown. We have demonstrated previously that in the human colonic epithelium, carcinoembryonic antigen (CEA) and CEA-related cell adhesion molecule 1 (CEACAM1) are integral components of the apical glycocalyx which participate in epithelial microbial interactions. In this study, based on the reactivity of specific monoclonal antibodies and on immunoelectron microscopy, we show that M cells of human colonic solitary lymphoid follicles express CEA and CEACAM1 on the apical surface. Recently these highly glycosylated molecules have been characterized as protein receptors for different bacteria. This leads us to propose a role for CEA and CEACAM1 in the adherence of enteric bacteria to the apical membrane of colonic M cells. We also hypothesize that, unlike colonic enterocytes, M cells lack the defense mechanism that eliminates CEA and CEACAM1 upon microbial binding and which is based on vesiculation of microvillus plasma membrane. PMID- 14758483 TI - Collagen and tenascin-C expression along the migration pathway of mouse primordial germ cells. AB - Primordial germ cells (PGCs) are the progenitor cells of the vertebrate germ line. These cells originate outside of the embryo and, through separation, migration, and colonization, arrive at the genital ridge, contributing to gonad development. Diverse extracellular matrix molecules are present along the PGC migratory pathway, permitting or inhibiting PGC displacement. Collagens and tenascin form the substratum for in vitro migration of neural crest cells and PGCs. However, little is known about the expression and distribution of these molecules during in situ PGC migration. Using immunohistochemistry, we identified tenascin-C and types I, III, and V collagen along the mouse PGC migration pathway. These molecules were spatiotemporally expressed in basement membranes of hindgut, coelomic epithelia, and mesonephric tubules and mesenchyme throughout the study. Our results complement previous data from our laboratory and contribute to building comprehension of the composition of the mouse PGC migratory pathway extracellular matrix, thereby enhancing understanding of the process. PMID- 14758484 TI - Stabilization of visual acuity with photodynamic therapy in eyes with chorioretinal anastomoses. AB - PURPOSE: (1) To evaluate, in a non-randomized, institutional, prospective study, the efficacy of photodynamic therapy with Visudyne (PDT) in neovascular age related macular degeneration (AMD) eyes with chorioretinal anastomoses (CRA). (2) To review, in a retrospective study and for comparison, the natural evolution of neovascular AMD eyes with CRA. METHODS: Prospective clinical and angiographic study of 17 consecutive eyes with CRA, treated with PDT. Retrospective clinical and angiographic study of the natural course of 17 consecutive patients with CRA. Masked best-corrected visual acuity (VA) and angiographic features at baseline and during the period of one year were evaluated. RESULTS: The two groups presented similar characteristics at baseline regarding age, sex, initial VA, duration of follow-up and angiographic features. PDT-treated eyes showed, at 1 year follow-up, VA stabilization or improvement in 73.3% of the eyes, no cases with very severe VA loss, and no fluorescein leakage in 46.6% of the eyes. In contrast, at 1-year follow-up the natural evolution of CRA was characterized by severe or very severe VA loss in 69% of the eyes and statistically significant mean VA loss (P=0.001) with persistence of fluorescein leakage in all cases. CONCLUSION: The natural history of AMD eyes with CRA leads to progressive and dramatic VA loss, which is associated with blindness in most of the cases. PDT with verteporfin can offer some benefit to these patients, allowing VA stabilization or improvement in more than two thirds of the cases, at one year. PMID- 14758486 TI - The GEDNAP blind trial concept part II. Trends and developments. AB - This article presents a review of the developments in the GEDNAP blind trials over the period covering the past 10 years (1993-2003), demonstrating the changing approach to DNA investigations in the European community as a whole. The results of the trials also identify the most common types of error encountered which can also occur during routine DNA typing and ways of recognising such errors are suggested. PMID- 14758485 TI - Digital 3D reconstruction of two parahissian accessory bundles in a case of Wolff Parkinson-White syndrome. AB - Three-dimensional reconstruction of digitized histological serial sections of the cardiac conduction system yielded two accessory pathways in a case of a 24-day old male infant who died after a short period of illness with ECG findings of Wolff-Parkinson-White syndrome. In infants, the differential diagnosis of possible accessory pathways connecting the AV conduction system, atrial or ventricular septum includes dispersed conduction system tissue without connecting features. This is why three-dimensional reconstruction is necessary in order to refute or establish connectivity of cell groups as found in histological serial slice images. PMID- 14758487 TI - Histological features of peripilar signs associated with androgenetic alopecia. AB - BACKGROUND: A study of the scalp in a large cohort of volunteers with androgenetic alopecia using macrophotographs showed the presence of peripilar signs (PPS) around the hair ostia. OBJECTIVE: The aim of the present study was to establish the histopathological features related to PPS. DESIGN: Prospective clinicopathological study. SETTING. Department of Dermatology, University Hospital of Bologna. PATIENTS: A group of 40 patients (21 males and 19 females) participated in the study. Macrophotographs of the scalp were taken using a Dermaphot camera and PPS were scored using a three-point scale. Hair density and PPS were clinically scored according to reference scales. Two punch biopsies from the photographed area were obtained from each subject and histological analysis was performed on vertical and horizontal sections. OBSERVATIONS: Clinical parameters indicated that PPS were already detectable on scalp with high hair density. Moreover, in patients with high hair density (score >4), a significant relationship was found between the PPS score and the global score for perifollicular infiltrates. Thus PPS are linked to superficial perifollicular lymphocytic infiltrates in early androgenetic alopecia. CONCLUSIONS: PPS could be the clinical signs reflecting the presence of perifollicular infiltrates. PMID- 14758488 TI - Vascularized nerve grafts for the treatment of large nerve gap after severe trauma to an upper extremity. AB - We reviewed the clinical outcomes of vascularized nerve grafts for the repair of large nerve gaps (longer than 20 cm) after severe trauma to an upper extremity. Six patients who underwent vascularized sural nerve grafting (five to the median nerve and one to the ulnar nerve) with a monitoring skin flap were evaluated. The length of the vascularized sural nerve grafts ranged from 20 to 30 cm, with a mean length of 23.3 cm. All but one of the monitoring skin flap grafts was successful. In those patients for whom the monitoring skin flap graft was successful, the mean static-2PD at the corresponding fingertip was 14.2 mm (range 10-20 mm). Evaluation of these patients with the Semmes-Weinstein test produced the following results: filament 6, two patients; filament 10, three patients. The results of this study show that vascularized sural nerve grafting should be considered as a clinical alternative for nerve reconstruction in patients with nerve defects longer than 20 cm. PMID- 14758489 TI - Double reverse-flow island flaps for two adjacent finger tissue defect. AB - INTRODUCTION: Soft-tissue reconstruction of fingertip injuries remains a challenge for hand surgery. Tissue loss of multiple digits is a serious problem for hand surgeons. Surgical possibilities include regional, distant and local flaps. In this study, five patients presented with tissue loss of two adjacent fingers and were treated by double reverse-flow island flaps. MATERIALS AND METHODS: The surgical technique is an application of the reverse-flow homodigital island flap for two adjacent finger tissue defects. Instead of one flap, double island flaps are applied to two adjacent finger tissue defects. The flaps are raised from the lateral or medial palmar surface of the proximal phalanx level. Anastomoses between the radial and ulnar digital arteries at the distal interphalangeal joint level are preserved. RESULTS: Three of the patients had tissue defects at the fingertip. In these cases, digital nerve anastomosis with the counter lateral digital nerve made the flaps sensitive. In two patients, the tissue defect was on the dorsum of the middle phalanx level. In these cases, the flaps were non-sensitive. Neither infection nor flap failure was seen in the patients. Sensitive function was satisfactory in fingertip applications. CONCLUSION: The reverse-flow homodigital island flap is a commonly used surgical technique for tissue defects in the fingers. The double reverse-flow island flaps involve the application of this technique for two adjacent fingers. The important point in the surgical technique is that the vascular supply of the two flaps should originate from the same common palmar digital artery. This technique offers a possibility to repair the defects of two adjacent fingers. PMID- 14758490 TI - Treatment of nonunions with irretrievable broken nail pieces in the distal fragment. AB - INTRODUCTION: We present our experience in the treatment of nonunited fractures of long bones in the presence of a retained fragment of the broken intramedullary nail. MATERIALS AND METHOD: Of 287 long bone fractures treated by intramedullary fixation, we recalled 5 patients with broken intramedullary nails. In two patients the distal fragments of broken nails were not extracted and refixation was performed by an Ilizarov ring fixation with the broken piece left in situ. RESULTS: The patients ambulated with full weight bearing immediately following surgery. We achieved good results in both patients with fracture union times between 16 and 22 weeks. No complication was observed. CONCLUSIONS: Ring fixation using thin wires or half pins inserted beside fragments of the broken nail provides a simple and minimally traumatic method of treatment in patients when encountering difficulties extracting a retained fragment of broken intramedullary nails. PMID- 14758491 TI - Repair of distal biceps tendon rupture with the Biotenodesis screw. AB - BACKGROUND: Distal biceps tendon ruptures are uncommon injuries with only around 300 cases reported in the literature. Current management tends to favour anatomical reinsertion of the tendon into the radial tuberosity, especially in young and active individuals. These injuries are commonly repaired using either a single anterior incision with suture anchors or the Boyd-Anderson dual incision technique. CASE REPORT: We report the use of a bioabsorbable interference screw for the repair of distal biceps tendon rupture using a minimal incision technique. In this technique the avulsed tendon and a bioabsorbable screw are secured in a drill hole on the radial tuberosity using whip stitch and fibre wire sutures according to Biotenodesis system guidelines. CONCLUSION: The technique described requires minimal volar dissection that is associated with a reduced number of synostosis and posterior interosseous nerve injuries. The bioabsorbable interference screw has all the advantages of being biodegradable and has been shown to have greater pullout strength than suture anchors. It is also a reasonable alternative to titanium screws in terms of primary fixation strength. The strong fixation provided allows early active motion and return to previous activities as seen in our case. PMID- 14758492 TI - Recipient non-hematopoietic bone marrow cells in the intestinal graft after fetal small intestinal transplantation. AB - We examined whether non-hematopoietic BM cells can migrate into the intestinal graft after fetal small intestinal transplantation (FSITx). Fetal small intestine from donor C57BL/6 mice was transplanted into the rectus abdominis of recipient C57BL/6 mice with only green fluorescent protein (GFP) BM cells (syngeneic FSITx). Intestinal grafts were harvested on days 5, 10, and 30 after FSITx and stained immunohistochemically using anti-CD45 antibody (a marker for hematopoietic BM cells). Although there were no GFP-positive cells identified in the epithelium of the graft intestinal villi, there were a few cells positive for both GFP and CD45 in the lamina propria on day 5 after FSITx, and many present on days 10 and 30. In some grafts there were only cells that were GFP positive/CD45 negative (i.e., non-hematopoietic BM cells) found in the lamina propria on days 10 and 30. These data indicate that non-hematopoietic BM cells as well as hematopoietic BM cells can migrate from the recipient's bone marrow, suggesting that recipient mesenchymal stem cells may be strongly implicated in graft regeneration and development after FSITx. PMID- 14758493 TI - Congenital prepubic sinus: is it a residual cloacal membrane and umbilicophallic groove? AB - We report two cases of congenital prepubic sinus (CPS) and discuss theories about its embryology and the etiology of its variants to improve the global understanding of this uncommon anomaly. Based on a review of the scant number of reported cases and our own experience, we postulate that CPS may be caused by a residual cloacal membrane and umbilicophallic groove and that its depth may determine the position of its ending. PMID- 14758494 TI - Transection of the innominate artery for tracheomalacia caused by persistent opisthotonus. AB - Patients with cerebral palsy often develop opisthotonus. The trachea may be pinched between the innominate artery and the cervical spine. This compartmentalized thoracic inlet results in severe tracheomalacia. We successfully released tracheal compression by transection of the innominate artery. In case 1, a 4-year-old girl with cerebral palsy and opisthotonus was admitted due to respiratory distress. Bronchoscopy revealed severe tracheomalacia 2 cm above the carina. An endotracheal stent was placed through a tracheostomy. Two months later, she developed tracheal bleeding and bronchoscopy demonstrated a trachea-innominate artery fistula. Magnetic resonance brain angiography showed the presence of Willis' circle, and transection of the innominate artery was justified. This was done through a low cervical skin incision. In case 2, a 6 year-old boy with cerebral palsy and opisthotonus had long-standing respiratory distress. Ventilatory support did not resolve the symptoms. The innominate artery was transected in the same fashion as in the first case. Case 1 has been free from respiratory distress for 4 months and case 2 for 3 years. Our experience suggests that the combination of tracheomalacia, opisthotonus causes severe respiratory distress. Transection of the innominate artery is a useful therapeutic strategy to release airway obstruction in this condition. PMID- 14758495 TI - Suspicion of prenatal pyriform sinus cyst and fistula: a case report. AB - There has been no report describing suspected prenatal pyriform sinus (PS) cyst and fistula. We report a case suspected by prenatal ultrasonography and fetal MRI. A large cystic mass was found in the left neck of the fetus. After the baby was born, preoperative laryngoscopic catheterization of the fistula tract was used to confirm the diagnosis and greatly facilitated the identification and excision of the PS cyst and fistula. PMID- 14758496 TI - Sigmoidocolocystoplasty for augmentation of iatrogenic small capacity bladder caused by direct injury to the bladder during inguinal hernia repair: long-term follow-up. AB - Inguinal hernia repair is the most common operation performed in pediatric surgical practice. However, this procedure can be difficult, even in the most experienced hands, and result in complications, especially in small infants. Injury to the bladder is one of the known complications of inguinal herniotomy, especially in infants less than 6 months old. We report the long-term follow-up of a case having bladder injury during inguinal hernia repair at the age of 3 months and at the age of 10 underwent sigmoidocolocystoplasty for augmentation of a small, contracted bladder and high-grade vesicoureteric reflux caused by the bladder injury. PMID- 14758497 TI - Intracorporeal electrohydraulic lithotripsy for intrahepatic bile duct stone formation after choledochal cyst excision. AB - An 18-year-old girl who had undergone excision of a choledochal cyst and Roux-en Y hepatico-jejunostomy at another hospital when 23 months old was referred to our department because of recurrent cholangitis. Radiological investigations showed stones lying in minimally dilated, right posterior intrahepatic bile ducts (IHBD). At laparotomy, the hepatico-jejunostomy site was incised, and a flexible endoscope inserted into the IHBD. Multiple stones packed in the IHBD were easily fragmented using an electro-hydraulic lithotripsy (EHL) device inserted through the endoscope, and removed. There were no EHL-related complications, and her postoperative progress was uneventful. She is currently well with no episodes of cholangitis after a follow-up period of 3 years. EHL is a simple, effective alternative method for removing IHBD stones after choledochal cyst excision. To the best of our knowledge, this is the first report of EHL being used to remove stones that developed in the IHBD after choledochal cyst excision. PMID- 14758498 TI - Acute torsion of the spleen: diagnosis and management. AB - We present a 2-year-old girl with a 24-hour history of abdominal pain, fever, and vomiting. The diagnosis of acute splenic torsion was made by means of color and power Doppler ultrasound. Management of this rare surgical emergency is discussed. PMID- 14758500 TI - Regulation of mitochondrial energy production in cardiac cells of rainbow trout (Oncorhynchus mykiss). AB - In skinned rat cardiac fibres, mitochondrial affinity for endogenous ADP generated by creatine kinase and Ca2+-activated ATPases is higher than for exogenous ADP added to the surrounding medium, suggesting that mitochondria are functionally coupled to creatine kinase and ATPases. Such a coupling may be weaker or absent in ectothermic vertebrate cardiac cells, because they typically have less elaborate intracellular membrane structures, higher glycolytic capacity and lower working temperature. Therefore, we examined skinned cardiac fibres from rainbow trout at 10 degrees C. The apparent mitochondrial affinity for endogenous ADP was obtained by stimulation with ATP and recording of the release of ADP into the surrounding medium. The apparent affinity for endogenous ADP was much higher than for exogenous ADP suggesting a functional coupling between mitochondria and ATPases. The apparent affinity for exogenous ADP and ATP was increased by creatine or an increase in Ca2+-activity, which should increase intrafibrillar turnover of ATP to ADP. In conclusion, ADP seems to be channelled from creatine kinase and ATPases to mitochondria without being released to the surrounding medium. Thus, despite difference in structure, temperature and metabolic capacity, trout myocardium resembles that of rat with regard to the regulation of mitochondrial respiration. PMID- 14758499 TI - The vagus and recurrent laryngeal nerves in experimental congenital diaphragmatic hernia. AB - BACKGROUND: The etiology of the anatomic and functional abnormalities of the esophagus in infants surviving congenital diaphragmatic hernia (CDH) remains unclear. We showed previously that fetal rats with CDH have malformations of neural crest-derived structures. The aim of this study was to examine the anatomy of the vagus and the recurrent laryngeal nerves, both of neural crest origin, in rats with CDH. METHODS: We used the nitrofen-induced CDH fetal rat model. Nine control fetuses from four dams and nine fetuses with CDH from seven dams were included in this study. Embryos were fixed in formalin and a thoracic block from the larynx to tracheal bifurcation was serially sectioned in the horizontal plane. One in every ten sections was stained with hematoxylin and eosin. The image was digitalized using biological software (TDR-3dbase). Vagus and recurrent laryngeal nerves, trachea, esophagus and the great vessels were examined. In order to obtain the three-dimensional reconstructions, 90-120 consecutive images were used. RESULTS: In comparison with controls there were striking abnormalities of the vagus and the recurrent laryngeal nerves in fetuses with CDH: (1) absence of the left (2/9) or right (2/9) vagus nerves; (2) absence of the left (3/9) or right (3/9) recurrent laryngeal nerves; (3) marked hypoplasia of the trunk of the vagus (2/9); (4) deviations of their normal course and change of normal anatomical relationships into the mediastinum (2/9); and (5) abnormal branching of the lower portion of the vagus (1/9). CONCLUSIONS: Rat fetuses with CDH have anomalies of the vagus and recurrent laryngeal nerves that support the concept of a neural crest involvement in the origin of this malformation. 3-D reconstructions allow a detailed analysis and provide a precise insight into the real anatomy. These observations may explain esophageal motility disorders in CDH. PMID- 14758501 TI - Sugar and protein digestion in flowerpiercers and hummingbirds: a comparative test of adaptive convergence. AB - Flowerpiercers are the most specialized nectar-feeding passerines in the Neotropics. They are nectar robbers that feed on the sucrose-rich diet of hummingbirds. To test the hypothesis that flowerpiercers have converged with hummingbirds in digestive traits, we compared the activity of intestinal enzymes and the gut nominal area of cinnamon-bellied flowerpiercers (Diglossa baritula) with those of eleven hummingbird species. We measured sucrase, maltase, and aminopeptidase-N activities. To provide a comparative context, we also compared flowerpiercers and hummingbirds with 29 species of passerines. We analyzed enzyme activity using both standard allometric analyses and phylogenetically independent contrasts. Both approaches revealed the same patterns. With the exception of sucrase activity, hummingbirds' digestive traits were indistinguishable from those of passerines. Sucrase activity was ten times higher in hummingbirds than in passerines. Hummingbirds and passerines also differed in the relationship between intestinal maltase and sucrase activities. Maltase activity was two times higher per unit of sucrase activity in passerines than in hummingbirds. The sucrase activity of D. baritula was much lower than that of hummingbirds, and not unlike that expected for a passerine of its body mass. With the exception of aminopeptidase-N activity, the digestive traits of D. baritula were not different from those of other passerines. PMID- 14758503 TI - Micropropagation of mature wych elm (Ulmus glabra Huds.). AB - Explants of mature vigorous donor trees of wych elm ( Ulmus glabra Huds.) that had not been previously exposed to Dutch elm disease were investigated for the influence of phytohormones and media on shoot multiplication rates and organogenic capacity. The regenerates were micropropagated from cultures that originated from 15-year-old progeny of plus trees. Two plus trees aged over 70 years showed recalcitrant responses. Thidiazuron in combination with 6 benzylaminopurine (BAP) induced a significantly higher number of shoots per explant than the most optimal BAP treatment (5.88 vs. 3.05 shoots). Woody plant medium and Dubovsky minimal medium had no significant effects on shoot formation and multiplication rates. All plantlets raised in vitro were phenotypically normal and successfully hardened to ex vitro conditions. Two experimental field plots with 3-year-old in vitro-propagated trees were established. PMID- 14758504 TI - Irradiated homologous costal cartilage: versatile grafting material for rhinoplasty. AB - For most surgeons, nasal septal cartilage is the first choice in septoplasty. However, when this source is depleted, an alloplastic implant material might be preferable over other autogenous donor sites in order to avoid additional scars, morbidity, and lengthened operating time. In the alloplastic spectrum, irradiated costal cartilage (ICC) has certain advantages. Herein, we present our results with ICC in a wide range of septorhinoplasties to show its versatility and reliability. Sixty-five patients were included in the study. There were 42 male and 23 female patients. According to the indications, there were four groups of patients: (I) secondary septorhinoplasty (n = 24), (II) traumatic deformity (n = 21), (III) primary septorhinoplasty (n = 13), (IV) deformity due to previous septal surgery (n = 7). The mean follow-up period was 33 months. No significant resorption was detected in any of the patients. Minor complications developed in four cases (6%), including deformity in the dorsal graft, excessive graft length, and erythematous nasal tips. Aesthetic and functional results were satisfactory in the remaining cases. The low incidence of major complications and the versatility of ICC make it a safe and reliable source of cartilage graft for both primary and secondary septorhinoplasties when autogenous septal cartilage is either insufficient or unsuitable. PMID- 14758508 TI - The Tubingen approach: identification, selection, and validation of tumor associated HLA peptides for cancer therapy. AB - There is substantial need for molecularly defined tumor antigens to prime cytotoxic T cells in vivo for cancer immunotherapy, especially in the case of tumor entities for which only a few tumor antigens have been defined so far. In this review, we present the "Tubingen approach" to identify, select, and validate large numbers of MHC/HLA class I-associated peptides derived from tumor associated antigens. Step 1 is the identification of naturally presented HLA associated peptides directly from primary tumor cells. Step 2 is selection of tumor-associated peptides from step 1 by differential gene expression analysis and data mining. Step 3 is validation of selected candidates by monitoring in vivo T-cell responses in the context of patient-individualized immunizations. Our approach combines methods from genomics, proteomics, bioinformatics, and T-cell immunology. The aim is to develop effective immunotherapeutics consisting of multiple tumor-associated epitopes in order to induce a broad and specific immune response against cancer cells. PMID- 14758509 TI - DMSA study performed during febrile urinary tract infection: a predictor of patient outcome? AB - Technetium-99m dimercaptosuccinic acid (DMSA) study has been advocated as a method for the assessment of renal sequelae after acute febrile urinary tract infection (UTI). However, it is not known whether DMSA scintigraphy performed during acute UTI has any prognostic value for outcome assessment. The objective of this study was to evaluate the usefulness of DMSA scintigraphy performed during UTI as a predictor of patient outcome, to identify children at risk of events [vesico-ureteral reflux (VUR) or recurrent UTI] that may lead to the development of progressive renal damage. One hundred and fifty-two children (including 78 girls) with a mean age of 20 months (range 1 month to 12 years) with first febrile UTI were evaluated by DMSA scintigraphy during acute UTI. After acute UTI, children were explored by voiding cysto-urethrography. Children who presented an abnormal DMSA study, or a normal DMSA study but VUR or recurrent UTI, underwent a DMSA control study 6 months after UTI. Children with VUR were followed up by direct radionuclide cystography. DMSA scintigraphy performed during acute UTI was normal in 112 children (74%). In 95 of these children, follow-up DMSA scintigraphy was not performed owing to a good clinical outcome. In the remaining 17 children, follow-up scintigraphy was normal. Forty children (26%) presented abnormal DMSA study during acute UTI. Twenty-five of them presented a normal follow-up DMSA, and 15 presented cortical lesions. Children with abnormal DMSA had a higher frequency of VUR than children with normal DMSA (48% vs 12%). It is concluded that children with normal DMSA during acute UTI have a low risk of renal damage. Children with normal follow-up DMSA and low grade VUR have more frequent spontaneous resolution of VUR. PMID- 14758510 TI - Improved tolerance to missing data in myocardial perfusion SPET using OSEM reconstruction. AB - When projection data are incomplete for various technical reasons, artefacts may occur in the reconstructed images. This study examines whether an iterative reconstruction method, the ordered subsets implementation of the EM algorithm (OSEM), can improve reconstruction and minimise the artefacts compared to filtered back-projection (FBP). We varied the number and location of projections removed to investigate when significant artefacts occur, and whether diagnosis is affected. Phantom studies were analysed with sequential orthogonal pairs of projection angles removed (as would typically occur when either data loss or severe motion is detected during acquisition with a right-angled, dual-head cardiac single-photon emission tomography system) and reconstructed with both FBP and OSEM. Twelve normal myocardial perfusion studies were also assessed to study the effect of missing projections on clinical diagnosis. Differences between reconstructions with intact versus missing data were measured. Also, reconstructed images were clinically assessed and scored on a five-point scale based on whether the artefacts would alter clinical interpretation. Although both reconstruction methods showed artefacts, the absolute differences between reconstructed phantom data with intact and missing projection sets were significantly greater (P<0.005) for FBP than for OSEM for all numbers of missing projections. The clinical data showed similar differences between FBP and OSEM reconstructions. The three observers noted superiority of OSEM compared to FBP, with reduced incidence of clinically significant artefacts. However, neither reconstruction method could tolerate six or more missing pairs from 32 projections. There was no significant dependence on the angular location of missing projections. In the absence of any attempt to correct for missing projections, OSEM reduced the influence of artefacts compared to FBP. PMID- 14758511 TI - Scientific production and impact of nuclear medicine in Europe: how do we publish? AB - We performed a bibliometric search covering a 1-year period to evaluate the number and the scientific "weight" of nuclear medicine papers published from European as compared with other countries. The scientific impact of our discipline was evaluated according to the impact factor of each publication, and we also aimed to identify those countries and topics that are making the principal contributions to the development of our discipline. To this end, a search on MEDLINE (PubMed) was run to find all peer-reviewed articles published between April 2002 and May 2003, using isotope definitions as search terms. A total of 3,292 publications were identified. Of these, 650 were of no nuclear medicine interest, 229 were reviews and 82 had no country specified. In absolute numbers, Europe leads research in nuclear medicine (939 papers, 38.9%) followed by the USA (608 papers, 25.2%). Among European countries, Germany is the nation that is currently making the greatest contribution to the scientific production of nuclear medicine in Europe. Articles concerning the use of nuclear medicine in oncology account for more than one-quarter of all published nuclear medicine papers. PMID- 14758512 TI - Early and late effects of coronary artery bypass grafting on cardiac haemodynamics during daily physical activities in patients with coronary artery disease. AB - This study investigated the early and late effects of coronary artery bypass grafting (CABG) on left ventricular (LV) function during different physical daily activities in patients with multivessel coronary artery disease (CAD). In ten patients with multivessel CAD, cardiac haemodynamics were assessed during a 6-min walk test and during stair climbing 8+/-2 days before (study 1) and 15+/-3 days (study 2) and 120+/-3 days (study 3) after CABG. LV function was monitored by an ambulatory radionuclide system. In study 1, the walk test induced a significant increase in relative end-diastolic volume (EDV) and end-systolic volume (ESV) and no change in LV ejection fraction as compared to rest. In both study 2 and study 3, EDV increased significantly and ESV was unchanged. As a consequence, LV ejection fraction rose from 48%+/-8% to 52%+/-10% and from 48%+/-7% to 51%+/-6%, respectively (both P<0.05). In study 1, stair climbing induced a significant increase in EDV and ESV and as a consequence LV ejection fraction decreased from 46%+/-8% to 42%+/-9% (P<0.05) as compared to rest. In both study 2 and study 3, EDV increased significantly whereas ESV did not change. As a consequence, LV ejection fraction rose from 48%+/-8% to 52%+/-7% and from 48%+/-8% to 51%+/-7%, respectively (both P<0.05). In conclusion, CABG has beneficial effects on cardiac performance during moderate or more intense physical activity in patients with multivessel CAD and these effects are due to improvement in systolic function. Radionuclide monitoring of LV function provides an objective method for quantitative evaluation of cardiac performance after CABG. PMID- 14758513 TI - Primary lung carcinoma metastatic to a solitary fibrous tumor. AB - We report the case of a 78-year-old man with a 2 month history of newly diagnosed metastatic lung adenocarcinoma, who presented with a left gluteal soft tissue mass. Histological examination of the mass revealed a solitary fibrous tumor containing metastases from adenocarcinoma. PMID- 14758514 TI - Intraosseous malignant peripheral nerve sheath tumor with local recurrence, lung metastases and death. AB - Malignant peripheral nerve sheath tumor (MPNST) is almost always seen in soft tissue. Skeletal involvement by MPNST is uncommon and usually results from secondary invasion. Primary MPNSTs are exceptionally rare. We report a surgically proven case of intraosseous MPNST, with local recurrence and lung metastasis during follow-up. The imaging and histological features of the case are described and the literature on the subject briefly reviewed. PMID- 14758515 TI - DNA isolation from soil samples for cloning in different hosts. AB - Many protocols to extract DNA directly from soil samples have been developed in recent years. We employed two extraction methods which differed in the method of lysis and compared these methods with respect to yield, purity and degree of shearing. The main focus was on the specific isolation of DNA from different microorganisms, especially DNA from actinomycetes, as these cells are very difficult to lyse, in contrast to non-actinomycetes. Thus, we used both methods to isolate DNA from Pseudomonas, Arthrobacter and Rhodococcus and from soil spiked with the respective microorganisms. Both methods rendered high DNA yields with a low degree of shearing, but differed in the type of cells that were lysed. By one protocol (utilizing enzymatic lysis) only DNA from the Gram-negative Pseudomonas strain could be obtained whereas, by the other protocol (utilizing mechanical lysis), all microorganisms that were used could be lysed and DNA extracted from them. Using a combination of both protocols, DNA from those organisms could be obtained selectively. Furthermore, one of the protocols was modified, resulting in higher DNA yield and purity. PMID- 14758516 TI - Monitoring the progress of infection and recombinant protein production in insect cell cultures using intracellular ATP measurement. AB - Several monitoring methods used to predict viable cell density have been the subject of extensive studies, including oxygen uptake rate, carbon dioxide evolution rate, optical density, NADH-dependent fluorescence and relative permittivity measurement. We propose intracellular ATP determination by bioluminescence assay to monitor the progress of baculovirus infection and recombinant protein production in insect cell cultures. We found that the ATP content in viable cells increased after virus addition. The increase in the ATP level was observed until the maximum recombinant protein accumulation was reached. At maximum product yield, the specific ATP content significantly decreased. Results obtained in both batch and fed-batch cultures demonstrated that the specific ATP level could be considered as a good indicator of recombinant protein productivity. Monitoring the cellular ATP content after viral infection makes it possible to define the optimum time for product harvest. The main advantage of applying the ATP assay as an index of the progress of infection and recombinant protein synthesis is its short time and sensitivity. PMID- 14758517 TI - Polyhydroxyalkanoate (PHA) granule formation in Ralstonia eutropha cells: a computer simulation. AB - Computer simulation of polyhydroxyalkanoate (PHA) granule formation in vivo could help to design strategies to optimize the fermentation process and achieve higher yields of PHA. It could also suggest biotechnological approaches to control the granule size and molecular weight of the polymer. A computer program simulating the formation of PHA granules inside a Ralstonia eutropha cell was developed, based on published experimental data. The results are applicable to R. eutropha cells or other microorganisms and transgenic plants, where polyhydroxybutyrate production is made possible by heterologous expression systems. The simulation starts at the outset of the PHA accumulation phase when the cells are small and contain no PHA granules. In the presence of abundant glucose, the cell responds to phosphorus limitation by producing 3-hydroxybutyryl-CoA which undergoes polymerization on the few PHA synthase molecules present in the cytoplasm. The amphiphilic PHA synthase-PHA complex attracts additional PHA synthase molecules and granules begin to grow from these initiation sites. Phosphorus limitation and the appearance of PHA in the cytoplasm also stimulate production of phasin molecules that attach themselves to the growing granules. As the granules grow bigger, they begin to touch each other and move to optimize their packing. The phasin coat prevents the granules from coalescing. The size of the cell increases and its prolate ellipsoid shape becomes closer to spherical. The accumulation process stops either when the supply of glucose is exhausted or when the granules become tightly packed within the cell, so that access to their surface is limited. All important variables, such as cell dimensions, granule size, counts of granule-associated molecules, PHA yield, degree of polymerization of the PHA molecules, etc., are recorded in real time during the simulation. Examples of virtual experiments with the cell and their results are shown. PMID- 14758518 TI - A food-grade delivery system for Lactococcus lactis and evaluation of inducible gene expression. AB - The genetic improvement of Lactococcus lactis is a matter of biotechnological interest in the food industry and in the pharmaceutical and medical fields. However, to construct a food-grade delivery system, both the presence of antibiotic markers or plasmid sequences should be avoided and the maintenance and expression of the cloned gene should be guaranteed. The objective of this work was to produce crossover mutants of L. lactis with a reporter gene under the control of an inducible promoter in order to evaluate the level of gene expression. We utilized a nuclease gene of Staphylococcus aureus as a reporter gene, P(nisA) as the nisin-inducible promoter, a non-essential gene involved in histidine biosynthesis of L. lactis as the site for homologous recombination, and pRV300 as a suicide vector for the genomic integration in L. lactis NZ9000. Single- and double-crossover mutants were identified by genotype and phenotype. Relative to episomal transformants of L. lactis, the level of expression of the heterologous protein after nisin induction was similar in the crossover mutants, suggesting that a single copy of the heterologous gene can be used to produce the protein of interest. PMID- 14758519 TI - Reductive transformation of methyl parathion by the cyanobacterium Anabaena sp. strain PCC7120. AB - Organophosphorus compounds are toxic chemicals that are applied worldwide as household pesticides and for crop protection, and they are stockpiled for chemical warfare. As a result, they are routinely detected in air and water. Methods and routes of biodegradation of these compounds are being sought. We report that under aerobic, photosynthetic conditions, the cyanobacterium Anabaena sp. transformed methyl parathion first to o,o-dimethyl o-p-nitrosophenyl thiophosphate and then to o,o-dimethyl o-p-aminophenyl thiophosphate by reducing the nitro group. The process of methyl parathion transformation occurred in the light, but not in the dark. Methyl parathion was toxic to cyanobacteria in the dark but did not affect their viability in the light. Methyl parathion transformation was not affected by mutations in the genes involved in nitrate reduction in cyanobacteria. PMID- 14758520 TI - Comparative analysis of genetic diversity and expression of amoA in wastewater treatment processes. AB - The genetic diversity and expression of amoA of autotrophic ammonia oxidizers in wastewater treatment processes were investigated by RT-PCR and denaturing gradient gel electrophoresis (DGGE) in order to identify active components of ammonia-oxidizer populations in a such processes. Ammonia oxidizers, evidenced by the presence of amoA mRNA, were regarded as metabolically active. The DGGE profiles derived from amoA mRNA and from its gene, which were amplified by RT-PCR or PCR using samples collected from a bench-scale reactor treating high concentration of inorganic ammonia, were similar. In contrast, RNA and DNA derived DGGE profiles from three domestic wastewater treatment facilities were different from each other. These data indicate that the dominant ammonia oxidizers in the bench-scale reactor exhibited ammonia-oxidizing activity, whereas some ammonia oxidizers in the domestic wastewater treatment facilities apparently did not express high levels of amoA mRNA. PMID- 14758521 TI - Construction of a self-cloning sake yeast that overexpresses alcohol acetyltransferase gene by a two-step gene replacement protocol. AB - The commercial application of genetically modified industrial microorganisms has been problematic due to public concerns. We constructed a "self-cloning" sake yeast strain that overexpresses the ATF1 gene encoding alcohol acetyltransferase, to improve the flavor profile of Japanese sake. A constitutive yeast overexpression promoter, TDH3p, derived from the glyceraldehyde-3-phosphate dehydrogenase gene from sake yeast was fused to ATF1; and the 5' upstream non coding sequence of ATF1 was further fused to TDH3p-ATF1. The fragment was placed on a binary vector, pGG119, containing a drug-resistance marker for transformation and a counter-selection marker for excision of unwanted DNA. The plasmid was integrated into the ATF1 locus of a sake yeast strain. This integration constructed tandem repeats of ATF1 and TDH3p-ATF1 sequences, between which the plasmid was inserted. Loss of the plasmid, which occurs through homologous recombination between either the TDH3p downstream ATF1 repeats or the TDH3p upstream repeat sequences, was selected by growing transformants on counter selective medium. Recombination between the downstream repeats led to reversion to a wild type strain, but that between the upstream repeats resulted in a strain that possessed TDH3p-ATF1 without the extraneous DNA sequences. The self-cloning TDH3p-ATF1 yeast strain produced a higher amount of isoamyl acetate. This is the first expression-controlled self-cloning industrial yeast. PMID- 14758522 TI - Aortic duplication artefact in a 14-year-old girl. PMID- 14758523 TI - When is recalcitrant pyoderma gangrenosum truly recalcitrant? AB - BACKGROUND: The literature supports many different therapies used in recalcitrant pyoderma gangrenosum, which suggests that the pathophysiology is not yet understood. Our hypothesis is that certain patient demographic factors, including age or size of ulcer at presentation, may predict a poor clinical outcome. OBJECTIVE: The goal of our study was to stimulate an analysis of pyoderma gangrenosum treatment failures to identify predisposing conditions that would increase the likelihood of failure. CONCLUSION: Patients older than 50 years of age or with an ulcer greater than 25 cm in diameter may have a poor outcome, but a systematic review of pyoderma gangrenosum treatment failures should be undertaken to support this hypothesis. PMID- 14758524 TI - Chloride ligation in inorganic manganese model compounds relevant to photosystem II studied using X-ray absorption spectroscopy. AB - Chloride ions are essential for proper function of the photosynthetic oxygen evolving complex (OEC) of Photosystem II (PS II). Although proposed to be directly ligated to the Mn cluster of the OEC, the specific structural and mechanistic roles of chloride remain unresolved. This study utilizes X-ray absorption spectroscopy (XAS) to characterize the Mn-Cl interaction in inorganic compounds that contain structural motifs similar to those proposed for the OEC. Three sets of model compounds are examined; they possess core structures Mn(IV)(3)O(4)X (X=Cl, F, or OH) that contain a di-micro-oxo and two mono-micro oxo bridges or Mn(IV)(2)O(2)X (X=Cl, F, OH, OAc) that contain a di-micro-oxo bridge. Each set of compounds is examined for changes in the XAS spectra that are attributable to the replacement of a terminal OH or F ligand, or bridging OAc ligand, by a terminal Cl ligand. The X-ray absorption near edge structure (XANES) shows changes in the spectra on replacement of OH, OAc, or F by Cl ligands that are indicative of the overall charge of the metal atom and are consistent with the electronegativity of the ligand atom. Fourier transforms (FTs) of the extended X-ray absorption fine structure (EXAFS) spectra reveal a feature that is present only in compounds where chloride is directly ligated to Mn. These FT features were simulated using various calculated Mn-X interactions (X=O, N, Cl, F), and the best fits were found when a Mn-Cl interaction at a 2.2-2.3 A bond distance was included. There are very few high-valent Mn halide complexes that have been synthesized, and it is important to make such a comparative study of the XANES and EXAFS spectra because they have the potential for providing information about the possible presence or absence of halide ligation to the Mn cluster in PS II. PMID- 14758525 TI - Iron inhibits neurotoxicity induced by trace copper and biological reductants. AB - The extracellular microenvironment of the brain contains numerous biological redox agents, including ascorbate, glutathione, cysteine and homocysteine. During ischemia/reperfusion, aging or neurological disease, extracellular levels of reductants can increase dramatically owing to dysregulated homeostasis. The extracellular concentrations of transition metals such as copper and iron are also substantially elevated during aging and in some neurodegenerative disorders. Increases in the extracellular redox capacity can potentially generate neurotoxic free radicals from reduction of Cu(II) or Fe(III), resulting in neuronal cell death. To investigate this in vitro, the effects of extracellular reductants (ascorbate, glutathione, cysteine, homocysteine or methionine) on primary cortical neurons was examined. All redox agents except methionine induced widespread neuronal oxidative stress and subsequent cell death at concentrations occurring in normal conditions or during neurological insults. This neurotoxicity was totally dependent on trace Cu (>or=0.4 microM) already present in the culture medium and did not require addition of exogenous Cu. Toxicity involved generation of Cu(I) and H(2)O(2), while other trace metals did not induce toxicity. Surprisingly, administration of Fe(II) or Fe(III) (>or=2.5 microM) completely abrogated reductant-mediated neurotoxicity. The potent protective activity of Fe correlated with Fe inhibiting reductant-mediated Cu(I) and H(2)O(2) generation in cell-free assays and reduced cellular Cu uptake by neurons. This demonstrates a novel role for Fe in blocking Cu-mediated neurotoxicity in a high reducing environment. A possible pathogenic consequence for these phenomena was demonstrated by abrogation of Fe neuroprotection after pre-exposure of cultures to the Alzheimer's amyloid beta peptide (Abeta). The loss of Fe neuroprotection against reductant toxicity was greater after treatment with human Abeta1-42 than with human Abeta1-40 or rodent Abeta1-42, consistent with the central role of Abeta1-42 in Alzheimer's disease. These findings have important implications for trace biometal interactions and free radical-mediated damage during neurodegenerative illnesses such as Alzheimer's disease and old-age dementia. PMID- 14758526 TI - Methionine-121 coordination determines metal specificity in unfolded Pseudomonas aeruginosa azurin. AB - Pseudomonas aeruginosa azurin binds copper so tightly that it remains bound even upon polypeptide unfolding. Copper can be substituted with zinc without change in protein structure, and also in this complex the metal remains bound upon protein unfolding. Previous work has shown that native-state copper ligands Cys112 and His117 are two of at least three metal ligands in the unfolded state. In this study we use isothermal titration calorimetry and spectroscopic methods to test if the native-state ligand Met121 remains a metal ligand upon unfolding. From studies on a point-mutated version of azurin (Met121Ala) and a set of model peptides spanning the copper-binding C-terminal part (including Cys112, His117 and Met121), we conclude that Met121 is a metal ligand in unfolded copper-azurin but not in the case of unfolded zinc-azurin. Combination of unfolding and metal titration data allow for determination of copper (Cu(II) and Cu(I)) and zinc affinities for folded and unfolded azurin polypeptides, respectively. PMID- 14758527 TI - Central neurotoxicity of cyclosporine in two children with nephrotic syndrome. AB - The central neurotoxicity of cyclosporin A (CsA) has been abundantly documented in pediatric and adult recipients of bone marrow or organ transplants, with variations in the rate of occurrence from 0.5% to 35%. We report two cases of central neurotoxicity ascribable to CsA in children with nephrotic syndrome due to lipoid nephrosis. The manifestations of CsA-related central neurotoxicity include confusion, aphasia, dystonias, akinetic mutism, parkinsonism, palsies, seizures, catatonia, coma, brain hemorrhage, and cortical blindness. Decreased density of the cerebral white matter is visible by computed tomography (CT) in 50% of cases, with the most commonly involved sites being the occipital cortex, the cerebellum, the periventricular substance, and the brainstem. Magnetic resonance imaging is more sensitive and more specific than CT for investigating the white matter. High-signal lesions are seen on T2-weighted sequences in the areas that are abnormal by CT. Many risk factors have been reported, including hypomagnesemia, hypocholesterolemia, high-dose glucocorticoid therapy, arterial hypertension, and infections. We present two patients with central neurotoxicity both of whom have elevated cholesterol levels. PMID- 14758528 TI - Tacrolimus therapy in pediatric patients with treatment-resistant nephrotic syndrome. AB - This is a retrospective analysis of 16 children started on tacrolimus with various types of treatment-resistant nephrotic syndrome. There are 13 patients with focal glomerulosclerosis, 1 minimal change disease, and 2 IgA nephropathy with nephrosis. The mean age of the children was 11.4 years (range 3.5-18.1 years) with a mean age at diagnosis of 5.6 years (range 1.6-13.3 years). All patients initially received prednisone 2 mg/kg per day. Other therapies for 15 of 16 included cyclosporine (n=15), chlorambucil (n=5), mycophenolate mofetil (n=5), levamisole (n=3), i.v. methylprednisolone (n=3), and cyclophosphamide (n=2). The major indication for the initiation of tacrolimus included treatment resistance/dependence (n=15) and intolerable side effects from other therapies (n=1). The average time from the diagnosis to initiation of tacrolimus was 5.3 years (range 0.3-13.3 years, median 6 years). The initial dosage of tacrolimus utilized was 0.1 mg/kg per day divided into two doses. The mean follow-up period was 6.5 months (range 2.5-18 months). Thirteen patients (81%) went into a complete remission within an average of 2 months (range 0.5-5.5 months), with 3 patients relapsing while on treatment. Three patients did not respond. Of these, 2 had partial remissions (13%) and 1 failed to respond. Adverse events included anemia (n=1), seizure (n=1), worsening or new-onset hypertension (n=5), and sepsis (n=1). All patients remain on tacrolimus. Tacrolimus is an effective, well tolerated medication for treatment-resistant forms of nephrotic syndrome in children, with a complete remission rate of 81% and a partial remission rate of 13% (totaling 94%). PMID- 14758529 TI - Osteopontin expression and microvascular injury in cyclosporine nephrotoxicity. AB - The aim of this study was to evaluate the role of osteopontin (OPN) in cyclosporine (CsA) nephrotoxicity of the human kidney. Renal biopsy samples obtained before and after 1-2 years of CsA treatment were evaluated in 18 children (2.2-13.0 years, 14 males, 4 females) diagnosed with minimal change nephrotic syndrome. The changes in tubular OPN expression between pre- and post treatment samples were correlated with interstitial macrophage infiltration, transforming growth factor-beta (TGF-beta) expression, interstitial fibrosis, and microvascular density. OPN, TGF-beta, CD68, and CD34 positivity were quantitatively assessed by immunohistochemical staining. Light microscopy showed that interstitial fibrosis developed in two-thirds of patients after CsA treatment. However, CD68-positive macrophages infiltrated minimally in fibrotic areas and were found in only one-third of patients. OPN expression was significantly increased in the glomerular mesangium (P=0.001) and tubules (P=0.025) after CsA treatment, whereas the number of CD34-positive peritubular capillaries decreased (P=0.022). An inverse relationship was observed between tubular OPN expression and microvascular density (r=-0.644). However, tubular OPN expression was not related to proteinuria, interstitial fibrosis, or interstitial or tubular TGF-beta expression. This study indicates that increased OPN expression may be related to microvascular injury in human CsA nephrotoxicity. It also shows that OPN expression may be used as an early but non-specific marker of CsA toxicity before the manifestation of interstitial fibrosis. PMID- 14758530 TI - Association of IL-1beta, IL-1ra, and TNF-alpha gene polymorphisms in childhood nephrotic syndrome. AB - We investigated the association between IL-1beta, IL-1ra, and TNF-alpha gene polymorphisms and childhood nephrotic syndrome (NS). We analyzed the genetic polymorphism of IL-1beta, IL-1ra, and TNF-alpha genes in 152 patients with childhood NS and 292 healthy adult controls. The C to T exchange at position -511 of IL-1beta and the G to A at -308 of the TNF-alpha gene were genotyped. Five alleles of the IL-1ra gene were identified and designated as IL1RN*1, IL1RN*2, IL1RN*3, IL1RN*4, and IL1RN*5, according to the variable number of tandem repeats in intron 2. The allele frequencies of IL-1beta1 (-511C), IL-1beta2 (-511T), TNF1 (-308G), and TNF2 (-308A) were 53.0, 47.0, 92.1, and 7.9%, respectively, in the childhood NS group. This was not significantly different from normal controls. In the childhood NS group, the allele frequencies of IL1RN*1, IL1RN*2, IL1RN*3, IL1RN*4, and IL1RN*5 were 90.8, 7.6, 1.6, 0, and 0% [IL1RN*1 odds ratio (OR)=0.296, P=0.0001, IL1RN*2 OR=3.902, P=0.0002]. A high allele frequency of IL1RN*2 and a lower allele frequency of IL1RN*1 were found in childhood NS, although there was no association with IL-1beta and TNF-alpha. A high allele frequency of the IL1RN*2 allele may affect disease susceptibility in childhood NS. PMID- 14758531 TI - A primer on the study of transitory dynamics in ecological series using the scale dependent correlation analysis. AB - Here we describe a practical, step-by-step primer to scale-dependent correlation (SDC) analysis. The analysis of transitory processes is an important but often neglected topic in ecological studies because only a few statistical techniques appear to detect temporary features accurately enough. We introduce here the SDC analysis, a statistical and graphical method to study transitory processes at any temporal or spatial scale. SDC analysis, thanks to the combination of conventional procedures and simple well-known statistical techniques, becomes an improved time-domain analogue of wavelet analysis. We use several simple synthetic series to describe the method, a more complex example, full of transitory features, to compare SDC and wavelet analysis, and finally we analyze some selected ecological series to illustrate the methodology. The SDC analysis of time series of copepod abundances in the North Sea indicates that ENSO primarily is the main climatic driver of short-term changes in population dynamics. SDC also uncovers some long-term, unexpected features in the population. Similarly, the SDC analysis of Nicholson's blowflies data locates where the proposed models fail and provides new insights about the mechanism that drives the apparent vanishing of the population cycle during the second half of the series. PMID- 14758532 TI - Patterns of nitrogen accumulation and cycling in riparian floodplain ecosystems along the Green and Yampa rivers. AB - Patterns of nitrogen (N) accumulation and turnover in riparian systems in semi arid regions are poorly understood, particularly in those ecosystems that lack substantial inputs from nitrogen fixing vegetation. We investigated sources and fluxes of N in chronosequences of riparian forests along the regulated Green River and the free-flowing Yampa River in semi-arid northwestern Colorado. Both rivers lack significant inputs from N-fixing vegetation. Total soil nitrogen increased through time along both rivers, at a rate of about 7.8 g N m(-2) year( 1) for years 10-70, and 2.7 g N m(-2)year(-1) from years 70-170. We found that the concentration of N in freshly deposited sediments could account for most of the soil N that accumulated in these floodplain soils. Available N (measured by ion exchange resin bags) increased with age along both rivers, more than doubling in 150 years. In contrast to the similar levels of total soil N along these rivers, N turnover rates, annual N mineralization, net nitrification rates, resin N, and foliar N were all 2-4 times higher along the Green River than the Yampa River. N mineralization and net nitrification rates generally increased through time to steady or slightly declining rates along the Yampa River. Along the Green River, rates of mineralization and nitrification were highest in the youngest age class. The high levels of available N and N turnover in young sites are not characteristic of riparian chronosequences and could be related to changes in hydrology or plant community composition associated with the regulation of the Green River. PMID- 14758533 TI - Detection of conspecific alarm cues by juvenile salmonids under neutral and weakly acidic conditions: laboratory and field tests. AB - A variety of fishes possess damage-released chemical alarm cues, which play a critical role in the detection and avoidance of potential predation threats. Recently, we have demonstrated that the ability of fathead minnows ( Pimephales promelas) and finescale dace ( Phoxinus neogaeus) to detect and respond to conspecific alarm cues is significantly reduced under weakly acidic conditions (pH 6.0). Rainbow trout ( Oncorhynchus mykiss) and brook charr ( Salvelinus fontinalis) possess an analogous alarm cue system. However, it is unknown if the trout alarm cue system is likewise affected by relatively small changes in pH. In addition, previous studies have not verified this phenomenon under natural conditions. We conducted laboratory and field trials to examine the potential effects of acute exposure to weakly acidic (pH 6.0) conditions on the detection and response of conspecific alarm cues by juvenile trout. Our laboratory results demonstrate that while juvenile rainbow trout exhibit significant increases in antipredator behaviour under normal pH conditions (pH 7.0-7.2), they do not respond to the presence of conspecific chemical alarm cues (i.e. response is not different from controls) under weakly acidic conditions. Similarly, a wild strain of brook charr in their natural streams near Sudbury, Ontario, failed to detect conspecific alarm cues in a weakly acidic stream (mean pH 6.11) while they responded to these cues in a neutral stream (mean pH of 6.88). This is the first demonstration that relatively small changes in ambient pH can influence alarm responses under natural conditions. These data suggest significant, sub-lethal effects of acid precipitation on natural waterways. PMID- 14758534 TI - Competitive effect versus competitive response of invasive and native wetland plant species. AB - Non-native plants can have adverse effects on ecosystem structure and processes by invading and out-competing native plants. I examined the hypothesis that mature plants of non-native and native species exert differential effects on the growth of conspecific and heterospecific seedlings by testing predictions that (1) invasive vegetation has a stronger suppressive effect on seedlings than does native vegetation, (2) seedlings of invasive species are better able to grow in established vegetation than are native seedlings, and (3) invasive species facilitate conspecific and inhibit heterospecific seedling growth. I measured growth rates and interaction intensities for seedlings of four species that were transplanted into five wetland monoculture types: invasive Lythrum salicaria; native L. alatum, Typha angustifolia, T. latifolia; unvegetated control. Invasive L. salicaria had the strongest suppressive effect on actual and per-individual bases, but not on a per-gram basis. Seedlings of T. latifolia were better able to grow in established vegetation than were those of L. salicaria and T. angustifolia. These results suggest that L. salicaria is not a good invader of established vegetation, but once established, it is fairly resistant to invasion. Thus, it is likely that disturbance of established vegetation facilitates invasion by L. salicaria, allowing it to compete with other species in even-aged stands where its high growth rate and consequent production of aboveground biomass confer a competitive advantage. PMID- 14758535 TI - Rapid development of phosphorus limitation in temperate rainforest along the Franz Josef soil chronosequence. AB - The aim of this study was to examine how shifts in soil nutrient availability along a soil chronosequence affected temperate rainforest vegetation. Soil nutrient availability, woody plant diversity, composition and structure, and woody species leaf and litter nutrient concentrations were quantified along the sequence through ecosystem progression and retrogression. In this super-wet, high leaching environment, the chronosequence exhibited rapid soil development and decline within 120000 years. There were strong gradients of soil pH, N, P and C, and these had a profound effect on vegetation. N:P(leaf) increased along the chronosequence as vegetation shifted from being N- to P- limited. However, high N:P(leaf) ratios, which indicate P-limitation, were obtained on soils with both high and low soil P availability. This was because the high N-inputs from an N fixing shrub caused vegetation to be P-limited in spite of high soil P availability. Woody species nutrient resorption increased with site age, as availability of N and P declined. Soil P declined 8-fold along the sequence and P resorption proficiency decreased from 0.07 to 0.01%, correspondingly. N resorption proficiency decreased from 1.54 to 0.26%, corresponding to shifts in mineralisable N. Woody plant species richness, vegetation cover and tree height increased through ecosystem progression and then declined. During retrogression, the forest became shorter, more open and less diverse, and there were compositional shifts towards stress-tolerant species. Conifers (of the Podocarpaceae) were the only group to increase in richness along the sequence. Conifers maintained a lower N:P(leaf) than other groups, suggesting superior acquisition of P on poor soils. In conclusion, there was evidence that P limitation and retrogressive forests developed on old soils, but N limitation on very young soils was not apparent because of inputs from an abundant N-fixing shrub. PMID- 14758536 TI - An overview and synopsis of techniques for directing stem cell differentiation in vitro. AB - The majority of studies on stem cell differentiation have so far been based in vivo, on live animal models. The usefulness of such models is limited, since it is much more technically challenging to conduct molecular studies and genetic manipulation on live animal models compared to in vitro cell culture. Hence, it is imperative that efficient protocols for directing stem cell differentiation into well-defined lineages in vitro are developed. The development of such protocols would also be useful for clinical therapy, since it is likely that the transplantation of differentiated stem cells would result in higher engraftment efficiency and enhanced clinical efficacy, compared to the transplantation of undifferentiated stem cells. The in vitro differentiation of stem cells, prior to transplantation in vivo, would also avoid spontaneous differentiation into undesired lineages at the transplantation site, as well as reduce the risk of teratoma formation, in the case of embryonic stem cells. Hence, this review critically examines the various strategies that could be employed to direct and control stem cell differentiation in vitro. PMID- 14758537 TI - Clinical, histopathologic, and genetic investigation in two large families with dentinogenesis imperfecta type II. AB - Dentinogenesis imperfecta (DI) type II, an inherited disorder affecting dentin, has been linked to mutations in the dentin sialophosphoprotein ( DSPP) gene on chromosome 4q21. The gene product is cleaved into two dentin-specific matrix proteins, dentin sialoprotein (DSP) and dentin phosphoprotein. The aim of this investigation was to study genotypes and phenotypes in two affected families with special reference to clinical, radiographic, and histopathologic manifestations. Seven affected members of Family A and five of Family B were documented clinically and radiographically; 14 and 10 teeth, respectively, were available for histopathologic investigation and prepared for ground sections, which were assessed semiquantitatively for dysplastic manifestations in the dentin according to the scoring system, dysplastic dentin score (DDS). Venous blood samples were collected from six affected and ten unaffected members of Family A, and from eight affected and six unaffected members of Family B. Genomic DNA was extracted and used for sequence analyses. The two families presented with different missense mutations. An Arg68Trp missense mutation in the DSP part of the gene was revealed in all six analyzed affected individuals in Family A. This mutation was not present in any of the ten healthy members. In Family B, an Ala15Val missense mutation involving the last residue of the signal peptide was found in all eight affected but in none of the six healthy members. The clinical and radiographic disturbances and DDS were more severe in Family B. The data indicate the presence of a genotype-phenotype correlation in DI type II. PMID- 14758538 TI - A study of the distributional characteristics of FMR1 transcript levels in 238 individuals. AB - Fragile X syndrome, the most common form of inherited mental retardation, is caused by hyperexpansion and hypermethylation of a CGG repeat tract in the 5' untranslated region of the FMR1 gene. This methylation causes the gene to be transcriptionally silenced. In addition to the common allele form with less than 41 repeats, there are two other allelic forms of the FMR1 gene that are unmethylated: premutation (61-200 CGG repeats) and intermediate (41-60 CGG repeats). Recently, premutation-specific phenotypes not related to fragile X syndrome have been reported: a 20-fold increased risk for premature ovarian failure (POF) among female carriers and an increased risk for a tremor ataxia syndrome (TAS) primarily among older male carriers. At the molecular level, increased levels of FMR1 transcript have been observed among premutation carriers. Increased levels of transcript may be causally related to the POF or TAS phenotypes or may be a surrogate of some other allelic property. In this report, we have examined the distributional properties of transcript levels by repeat size and gender among 238 individuals. We have confirmed a significant linear relationship between transcript level and repeat size in males and females. The evidence for the linear effect is primarily within the premutation size alleles. PMID- 14758540 TI - An insertional mutation in the rice PAIR2 gene, the ortholog of Arabidopsis ASY1, results in a defect in homologous chromosome pairing during meiosis. AB - To elucidate the genetic system that establishes homologous chromosome pairing in monocot plants, we have isolated an asynaptic mutant of rice, designated pair2 (homologous pairing aberration in rice meiosis 2), in which 24 completely unpaired univalents are observed at pachytene and diakinesis. The mutation was caused by an insertion of the retrotransposon Tos17, as demonstrated by complementation of the mutation by transformation with the corresponding wild type gene. The gene in which the element was inserted is orthologous to the ASY1 gene of Arabidopsis thaliana and the HOP1 gene of Saccharomyces cerevisiae. Mature PAIR2 mRNA and several splicing variants were found to be highly expressed in wild-type reproductive tissues, and lower expression was also detected in vegetative tissues. In situ hybridization and BrdU incorporation experiments revealed that PAIR2 expression is specifically enhanced in male and female meiocytes, but not in those at pre-meiotic S phase or in the pollen maturation stages. The results obtained in this study suggest that the PAIR2 gene is essential for homologous chromosome pairing in meiosis, as in the case of the genes ASY1 and HOP1. The study also suggested the possibility that a highly homologous copy of the PAIR2 gene located on a different chromosome is in fact a pseudogene. PMID- 14758541 TI - The transcription factor Pap1/Caf3 plays a central role in the determination of caffeine resistance in Schizosaccharomyces pombe. AB - We previously identified four nuclear genes (caf1+-caf4+) in Schizosaccharomyces pombe, mutations in which confer resistance to caffeine and brefeldin A. caf1+, caf2+ and caf4+ were sequenced and found to be identical to the multidrug resistance/stress-response genes hba1, crm1 and trr1, respectively. Here we show that caf3 is allelic to pap1, which encodes an AP-1-like transcription factor. The allele associated with caffeine resistance, caf3-89, contains a single nucleotide exchange that results in a Leu-->Ser exchange in the NES (nuclear export signal) domain of the gene product. Due to this alteration, the modified protein can not be exported from the nucleus back into the cytoplasm, and thus accumulates in the nucleus. The activity of pap1/caf3 is shown to be necessary for manifestation of the caffeine resistance caused by mutations in the genes hba1/caf1 and crm1/caf2. We also cloned two genes that confer caffeine resistance when carried on a multicopy plasmid. One of them turned out to be a truncated allele of pad1/bfr2/sks1, which codes for a subunit of the 26 S proteosome. The putative product of the other gene, designated caf5, has a structure highly similar to that of MFS permeases. It contains two groups of six transmembrane spanning domains each, with the conserved motifs WRW, PET and GAIGGPVLGP in the fifth and sixth domains. These results are all consistent with our earlier hypothesis, which suggested that the caf genes are functionally interlinked in a complex detoxification mechanism. caf5 and pad1 may also encode parts of this mechanism. PMID- 14758542 TI - The roles of different regions of the CycH protein in c-type cytochrome biogenesis in Sinorhizobium meliloti. AB - Cytochrome c heme lyases encoded by the Sinorhizobium meliloti cycHJKL operon are responsible for generating the covalent bond between the heme prosthetic group and apocytochromes c. The CycH protein with its presumably membrane-associated N terminal and periplasmic C-terminal parts is thought to be responsible for binding apocytochrome and presenting it to the heme ligation machinery. We propose that these two modules of CycH play roles in different functions of the protein. The N-terminal 96 amino acids represent an active subdomain of the protein, which is able to complement the protoporphyrin IX (PPIX) accumulation phenotype of the cycH mutant strain AT342, suggesting that it is involved in the final steps of heme C biosynthesis. Furthermore, three tetratricopeptide (TPR) domains have been identified in the C-terminal periplasmic region of the CycH protein. TPR domains are known to mediate protein-protein interactions. Each of these CycH domains is absolutely required for protein function, since plasmid constructs carrying cycH genes with in-frame TPR deletions were not able to complement cycH mutants for their nitrate reductase (Rnr-) and nitrogen-fixing (Fix-) phenotypes. We also found that the 309-amino acid N-terminal portion of the CycH, which includes all the TPR domains, is able to mediate the assembly of the c-type cytochromes required for the Rnr+ phenotype. In contrast, only the full-length protein confers the ability to fix nitrogen. PMID- 14758544 TI - Clinical response to antibiotic therapy for community-acquired pneumonia. AB - Childhood community-acquired pneumonia is a common and potentially serious problem worldwide. Unless the patient has bacteraemia or pleural empyema, aetiological diagnostics are limited and antibiotic treatment is empirical. Published data on expected response to therapy are scarce. To determine the clinical response to antibiotic treatment in a developed country in otherwise healthy children with community-acquired pneumonia, we conducted a prospective study of 153 hospitalised children with pneumonia. The role of 17 microbes as potential causative agents was evaluated. The duration of fever (>37.5 degrees C) and hospitalisation were studied as objective measures of recovery. A potential aetiology was found in 83% of 153 patients: 29% of the patients had sole viral and 26% sole bacterial and 29% mixed viral-bacterial infections. The median duration of fever after the onset of antibiotic treatment (mainly penicillin G) was 14 h and the median duration of hospitalisation was 48 h. Patients with mixed viral-bacterial infection became afebrile more slowly than those with either sole viral or sole bacterial infections. CONCLUSION: the findings indicate that in a developed country, children with pneumonia make a rapid, uneventful recovery needing only a short hospital stay. Expensive and time-consuming microbiological investigations are not required once bacterial sepsis has been excluded. PMID- 14758543 TI - Sources and predictors of resolvable indel polymorphism assessed using rice as a model. AB - The principal sources of genetic variation that can be assayed with restriction enzymes are base substitutions and insertions/deletions (indels). The likelihood of detecting indels as restriction fragment length polymorphisms (RFLPs) is determined by the size and frequency of the indels, and the ability to resolve small indels as RFLPs is limited by the distribution of restriction fragment sizes. In this study, we use aligned sequences from the indica and japonica subspecies of rice ( Oryza sativaL.) to quantify and compare the ability of restriction enzymes to detect indels. We look specifically at two abundant transposable element-derived indel sources: miniature inverted repeat transposable elements (MITEs) and long terminal repeat (LTR) retroelements. From this analysis we conclude that indels rather than base substitutions are the prevailing source of the polymorphism detected in rice. We show that, although MITE derived indels are more abundant than LTR-retroelement derived indels, LTR retroelements have a greater capacity to generate visible restriction fragment length polymorphism because of their larger size. We find that the variation in the detectability of indels among restriction enzymes can be explained by differences in the frequency and dispersion of their restriction sites in the genome. The parameters that describe the fragment size distributions obtained with the restriction enzymes are highly correlated across the sequenced genomes of rice, Arabidopsis and human, with the exception of some extreme deviations in frequency for particular recognition sequences corresponding to variations in the levels and modes of DNA methylation in the three disparate organisms. Thus, we can predict the relative ability of a restriction enzyme to detect indels derived from a specific source based on the distribution of restriction fragment sizes, even when this is estimated for a distantly related genome. PMID- 14758546 TI - TRK neurotrophin receptor-like proteins in the kidney of frog (Rana esculenta) and lizard (Podarcis sicula): an immunohistochemical study. AB - The present study investigates the occurrence of Trk-like neurotrophin receptor proteins in the lizard and frog kidney. In lizard rare TrkB-like immunoreactive cells in intermediate and distal tubules were found. TrkC-like immunoreactive cells were numerous in collecting tubules and became less numerous in collecting ducts. No TrkC-like immunoreactivity was detected in the ureteric duct. In the frog, we observed numerous TrkC-like immunoreactive cells in collecting tubules and ducts while they were scattered among negative epithelial cells in the wolffian duct. TrkB- and TrkA-like immunoreactivity was never found. Western blot analysis demonstrated that the frog and lizard kidney contains TrkC-like protein; TrkB-like protein was present only in the lizard kidney. These results demonstrate for the first time the occurrence of Trk-like proteins in the kidney of amphibians and reptiles, and aid in the assessment of the role of Trk receptor like proteins in the kidney physiology of vertebrates. PMID- 14758545 TI - Interaction of immune complexes isolated from hepatitis C virus-infected individuals with human cell lines. AB - We investigated the interaction of immune complexes (IC) isolated from hepatitis C virus (HCV)-infected individuals with several cell lines that differentially express Fc receptors, and analyzed viral infection by the presence of HCV RNA sequences. Monocytic (U937 and Monomac-6) and lymphocytic (MOLT-4 and Jurkat) cell lines were incubated with interferon- plus phorbol myristate acetate to stimulate the expression of Fc receptors before addition of IC. Cell interaction with IC was monitored by flow cytometry. Positive cell fluorescence was detected in U937 and Monomac-6 cells [mean fluorescence intensity (MFI) 10.56+/-0.8 and 11.60+/-0.8, respectively]. Incubation of cells with monoclonal antibodies against Fc receptors for IgG before addition of IC decreased MFI in both cell lines (U937 2.1+/-0.5, Monomac-6 4.4+/-0.8, P<0.001), indicating that cell-IC interaction through these receptors was inhibited. In particular, the blockage of FcgammaRII was responsible for this effect. No binding of IC with either MOLT-4 or Jurkat cell lines was detected, which correlated with a very low Fc receptor expression. HCV RNA sequences were identified in the cells up to 120 h of post incubation with IC. These results suggest that IC can mediate entry of HCV to both U-937 and Monomac-6 cell lines mainly through the FcgammaRII. PMID- 14758547 TI - Uncommon ureteric ectopias: embryological implications. AB - We herein report two rare ureteric ectopias, one in the uterus, the second in the rectum. These kinds of ectopias are not easily explained with regard to the classic embryological theories of the too-cranial or too-caudal origin of the ureteric bud on the mesonephric duct. We thus set out new explanations consistent with embryological studies on the narrow contact between the developing paramesonephric and mesonephric ducts, and on the so-called cloacal septation. PMID- 14758548 TI - Expression of kinesin kif5c during chick development. AB - Kinesins are molecular motors associated with microtubules. They act mainly as intracellular transport proteins carrying different cargos like organelles along the microtubules. We cloned the avian homologue of the mammalian kif5c gene, a member of the khc family coding for the heavy chain of conventional kinesin. Its murine homologue has been described to be specific for neuronal tissue. Here we present the expression pattern of kif5c in chick embryos. We found a highly dynamic expression pattern for kif5c in a variety of developing tissues including neuronal and mesodermal tissues. In young embryos the expression pattern around Hensen's node is asymmetric with stronger expression on the right side, implying that kif5c is involved in the formation of the left-right body axis. A connection with intracellular transport linked to early asymmetric morphogenesis in the node is likely. Vesicles containing signaling molecules could be possible cargos. At later stages, kif5c expression is found in the paraxial, intermediate and somatic mesoderm and in the tail bud. The expression in the paraxial mesoderm occurs first during segmentation and continues in the epithelial somites and the dermomyotome. During neurulation kif5c is expressed in ectodermal and neural plate cells. In older embryos, the expression is restricted to the dorsal root and cranial ganglia, neural tube and olfactory tract. Taken together, our results demonstrate that in the chick embryo, kif5c plays a role during different morphogenetic processes. PMID- 14758549 TI - Expression and regulation of ROR-1 during early avian limb development. AB - ROR-1 is a member of the ROR family of tyrosine kinase like orphan receptors and is highly conserved among various species. We have isolated the chick ROR-1 ( cROR-1) and show that cROR-1 expression is high and restricted to the proximal limb region until HH-stage 25. At later stages, expression spreads towards the distal limb region. In order to determine the signals that control cROR-1 expression, factors known to be involved in limb patterning (FGFs, BMPs, SHH, retinoic acid) were applied to the developing limb. Whereas neither FGFs, BMPs, nor SHH affected cROR-1 expression, upregulation could be achieved by ectopic application of retinoic acid to the distal limb region. As retinoic acid also upregulated retinoic acid receptor beta ( Rar-beta), we assume that cROR-1 upregulation is mediated by Rar-beta. We conclude that ROR-1 signaling is an independently regulated pathway, which is involved in late rather than early limb development. PMID- 14758550 TI - In situ expression of connective tissue growth factor in human crescentic glomerulonephritis. AB - Connective tissue growth factor (CTGF) has recently been recognized as an important profibrotic factor and is up-regulated in various renal diseases with fibrosis. The present study describes the sequential localization of CTGF mRNA and its association with transforming growth factor (TGF)-beta1 in human crescentic glomerulonephritis (CRGN). Furthermore, we examined the phenotype of CTGF-expressing cells using serial section analysis. Kidney biopsy specimens from 18 CRGN patients were examined using in situ hybridization and immunohistochemistry. CTGF mRNA was expressed in the podocytes and parietal epithelial cells (PECs) in unaffected glomeruli. In addition, it was strongly expressed in the cellular and fibrocellular crescents, particularly in pseudotubule structures. Serial sections revealed that the majority of CTGF mRNA positive cells in the crescents co-expressed the epithelial marker cytokeratin, but not a marker for macrophages. Moreover, TGF-beta1, its receptor TGF-beta receptor-I, and extracellular matrix molecules (collagen type I and fibronectin) were co-localized with CTGF mRNA-positive crescents. Our results suggest that CTGF is involved in extracellular matrix production in PECs and that it is one of the mediators promoting the scarring process in glomerular crescents. PMID- 14758551 TI - Confocal microscope analysis and tridimensional reconstruction of papillary thyroid carcinoma nuclei. AB - Nuclei of papillary thyroid carcinoma (PTC) are characterised by diagnostic morphological features, which include optically clear nuclei, irregular nuclear profile, pseudoinclusions and grooves. In the present study, such nuclear features were analysed by means of confocal microscopy using anti-lamin B antibodies to outline the nuclear membrane. Parallel sections of the nucleus, produced by confocal microscope analysis, showed that the nuclear shape is markedly irregular with profound invaginations, clefts and tunnel-like structures, which correspond to the "grooves" and "holes" detectable using light microscopy, respectively. A tridimensional (3-D) model of the nuclei, obtained by a computer-based reconstruction of confocal microscope images, showed, in the vast majority of PTC cells, nuclei with crateriform areas, clefts and even tunnel like structures "piercing" the whole nuclear thickness. By rotating these models in space, it became evident that the holes and grooves seen in light microscopy correspond to invaginations and tunnels, depending on the viewpoint. In conclusion, this is the first application of confocal microscopy and tridimensional reconstruction to the study of nuclear morphology of PTC and of tumours in general. The light microscopic appearance of PTC nuclei, so familiar to pathologists, is, therefore, due to profound remodelling of the nuclear shape with invaginations and tunnels, which appear as either grooves or holes, according to the viewpoint. PMID- 14758552 TI - CD34+ fibrocytes, alpha-smooth muscle antigen-positive myofibroblasts, and CD117 expression in the stroma of invasive squamous cell carcinomas of the oral cavity, pharynx, and larynx. AB - We investigated tumor-free mucosa and squamous cell carcinomas of the oral cavity, the pharynx, and larynx with respect to the presence of stromal CD34+ fibrocytes and alpha-smooth muscle antigen (SMA)-positive myofibroblasts. Additionally, stromal expression of CD117 was analyzed. A total of 39 squamous cell carcinomas were assessed immunohistochemically. In all cases investigated, CD34+ fibrocytes were found in the tumor-free stroma, whereas alpha-SMA-positive myofibroblasts were lacking. Areas of lymphocytic infiltration disclosed a focal reduction of CD34+ fibrocytes. CD117 expression was absent from the tumor-free stroma. Of 39 squamous cell carcinomas, 33 were free of stromal CD34+ fibrocytes, and, in 31 carcinomas, stromal alpha-SMA-positive myofibroblasts occurred at least focally. CD117-positive stromal spindle cells were found in 25 carcinomas. Compared with tumor-free mucosa, the number of tissue mast cells was significantly increased in carcinomas. We conclude that stromal remodeling induced by invasive carcinomas is characterized by a loss of CD34+ fibrocytes and subsequent gain of alpha-SMA-positive myofibroblasts. The diagnostic impact of this finding is, however, limited by the fact that chronic inflammation may also be accompanied by a focal loss of CD34+ fibrocytes. PMID- 14758553 TI - MUC5B expression in gastric carcinoma: relationship with clinico-pathological parameters and with expression of mucins MUC1, MUC2, MUC5AC and MUC6. AB - Previous studies have shown that mucin expression can be used to evaluate differentiation patterns of gastric carcinoma: MUC5AC expression is associated with diffuse type and early gastric carcinomas, and MUC2 expression is associated with mucinous gastric carcinomas. The role played by MUC5B in the evaluation of differentiation and biological behaviour of gastric carcinoma is largely unknown. Our aim was to characterise the pattern of expression of mucins MUC1, MUC2, MUC5AC, MUC5B and MUC6 in a series of 50 gastric carcinomas to evaluate whether MUC5B expression was associated with the clinico-pathological characteristics of the cases and/or with the co-expression of other mucins. A panel of six monoclonal antibodies (HMFG1, SM3, PMH1, CLH2, EU-MUC5Ba and CLH5) was used to determine the expression of mucins (MUC1, MUC1 underglycosylated form, MUC2, MUC5B, MUC5AC and MUC6, respectively) using immunohistochemistry. Cases were considered positive if more than 5% of the cells expressed immunoreactivity for the several mucins evaluated. Our results showed that: (a) expression of MUC5B was observed in 11 cases (22.0%) and was associated with the "unclassified" histological type of gastric carcinoma according to Lauren ( P = 0.03) and with the absence of venous invasion ( P = 0.02); (b) in this series, MUC5B expression had no impact on survival of patients with gastric carcinoma; (c) the expression of MUC5B was associated with the co-expression of MUC5AC ( P = 0.02) and (d) none of the cases with the so-called complete intestinal phenotype of mucin expression expressed MUC5B. PMID- 14758554 TI - Analysis of Fz10 expression in mouse embryos. AB - Wnt signaling molecules regulate the development of multiple organs in vertebrate embryos. We have isolated cDNA clones for frizzled10 (Fz10), which encodes a putative Wnt receptor, to further characterize the mechanisms of Wnt signaling in mouse embryos. Interestingly, Fz10 is expressed in the same regions as Wnt7a in the neural tube, limb buds, and Mullerian duct. PMID- 14758555 TI - A growth kinetic model of Kluyveromyces marxianus cultures on cheese whey as substrate. AB - This work presents a multi-route, non-structured kinetic model for determination of microbial growth and substrate consumption in an experimental batch bioreactor in which beta-galactosidase is produced by Kluyveromyces marxianus growing on cheese whey. The main metabolic routes for lactose, and oxygen consumption, cell growth, and ethanol production are derived based on experimental data. When these individual rates are combined into a single growth rate, by rewriting the model equations, the model re-interpretation has a complexity similar to that of the usual variations of the Monod kinetic model, available in the literature. Furthermore, the proposed model is in good agreement with the experimental data for different growth temperatures, being acceptable for dynamic simulations, processes optimization, and implementations of model-based control technologies. PMID- 14758556 TI - Purification and characterisation of alkaline cellulase produced by a novel isolate, Bacillus sphaericus JS1. AB - A novel strain of Bacillus sphaericus JS1 producing thermostable alkaline carboxymethyl cellulase (CMCase; endo-1,4-beta-glucanase, E.C. 3.2.1.4) was isolated from soil using Horikoshi medium at pH 9.5. CMCase was purified 192-fold by (NH(4))(2)SO(4) precipitation, ion exchange and gel filtration chromatography, with an overall recovery of 23%. The CMCase is a multimeric protein with a molecular weight estimated by native-PAGE of 183 kDa. Using SDS-PAGE a single band is found at 42 kDa. This suggests presence of four homogeneous polypeptides, which would differentiate this enzyme from other known alkaline cellulases. The activity of the enzyme was significantly inhibited by bivalent cations (Fe(3+) and Hg(2+), 1.0 mM each) and activated by Co(2+), K(+) and Na(+). The purified enzyme revealed the products of carboxymethyl cellulose (CMC) hydrolysis to be CM glucose, cellobiose and cellotriose. Thermostability, pH stability, good hydrolytic capability, and stability in the presence of detergents, surfactants, chelators and commercial proteases make this enzyme potentially useful in laundry detergents. PMID- 14758557 TI - Biodecolourisation of some industrial dyes by white-rot fungi. AB - Eight white-rot fungal strains were screened for biodecolourisation of eight dyes commercially employed in various industries. Decolourisation of Poly R 478 was used as a standard to ascertain the dye-decolourisation potential of various fungi. All the fungi tested significantly decolourised Poly R 478 on solid agar medium. When tested in a nitrogen-limited broth medium, Dichomitus squalens, Irpex flavus, Phlebia spp. and Polyporus sanguineus were better industrial dye decolourisers than Phanerochaete chrysosporium. PMID- 14758558 TI - 47th annual meeting of the Society for Research into Hydrocephalus and Spina Bifida. Baltimore, Maryland, USA, 2003. Proceedings and abstracts. PMID- 14758559 TI - Flow control versus antisiphon valves: late results concerning slit ventricles and slit-ventricle syndrome. AB - INTRODUCTION: The aims of the study were firstly to investigate the time course of development of slit ventricles (SV) and slit-ventricle syndrome (SVsyndrome) in hydrocephalic patients shunted as infants and secondly to assess the difference in incidence of SV and SVsyndrome in 2 groups of patients - group A with a conventional valve and group B with an anti-siphon valve. PATIENTS AND METHODS: 27 infant patients with hydrocephalus received a ventriculoperitoneal shunt and were followed prospectively with annual clinic visits and at least two CT or MRI scans postoperatively. The scans were assessed for SV, and ventricular and parenchymal surface and cortical mantle thickness were measured. Revisions for shunt malfunction were recorded. RESULTS: SV were more frequent than SVsyndrome and developed within 6.5 months postoperatively in 21 % of patients and 48 % after 6 years. No significant difference in incidence of SV or in the surface and cortical mantle thickness were found between the two groups. Two patients (40 %) with early development of SV developed SVsyndrome, and the relative risk for shunt revision was significantly higher in patients who developed SV early. 48 % of patients did not need emergency shunt revision during the first 9 years. CONCLUSIONS: SV develop over years. The majority of patients with SV remain asymptomatic. No significant difference between the two groups was found concerning SV and SVsyndrome. If SV develop early in the postoperative period, complications are more frequent in the long-term, so elective valve adjustment should be considered. PMID- 14758560 TI - Long-term outcome in bladder detrusorectomy augmentation. AB - BACKGROUND AND OBJECTIVES: Bladder augmentation in children can have significant benefits in terms of continence and social function. In an attempt to overcome the problems associated with using foreign mucosa in the urinary tract, techniques to increase to bladder volume and improve compliance by resecting the detrusor muscle alone have been described. Here we describe our experience using an omental-backed detrusorectomy augmentation and examine possible reasons for a poor outcome observed in some patients in the longer term. METHODS: This was a retrospective review covering an eight-year period with a minimum follow-up of 2 years. Pre- and post-operative urodynamics were performed in every case. RESULTS: 11 patients were included in the study. There were 7 male and 4 female patients with a median age of 10 years (range 4 - 16). The underlying pathology in 6 patients was myelodysplasia and in the remaining cases there was one each of ano rectal malformation with neuropathic bladder, Guillain-Barre syndrome, a myopathy of unknown cause, transverse myelitis and one case of a non-neuropathic neuropathic bladder. Detrusorectomy provided a modest increase in bladder capacity (median 26 %) and decrease in maximum bladder pressure (median 12 %). Long-term follow-up has revealed treatment failure in 6 patients, resulting in revision augmentation surgery in 3 (with surgery planned in a further 2), and one patient developing end-stage renal failure. Notable complications were bladder stone formation in 4 patients and troublesome lower abdominal pain related to bladder drainage in 2. There appeared to be no correlation between initial diagnosis, age at operation, pre-operative urodynamics, peri- or post-operative factors, and long-term outcome. CONCLUSIONS: In our series, omental-backed detrusorectomy for a neuropathic bladder in children resulted in a poor outcome in 55 % of cases. We were unable to identify factors that would allow this result to be predicted pre-operatively. PMID- 14758561 TI - Chronic headaches in adults with spina bifida and associated hydrocephalus. AB - INTRODUCTION: Adults with spina bifida and associated hydrocephalus are exposed to multiple risk factors for the development of chronic headache. The management of these patients can be complex and misdiagnosis can precipitate unnecessary shunt revision. This study aims to evaluate the usefulness of intracranial pressure (ICP) monitoring as a diagnostic tool in these cases and to look at the causes of chronic headaches and treatment outcomes for this patient population. METHODS: All patients over the age of 18 years with a diagnosis of spina bifida and shunted hydrocephalus who had undergone inpatient or outpatient neurosurgical review within the last 10 years were identified in our hospital database. Case notes were then retrospectively reviewed to identify all patients who had undergone either inpatient or outpatient evaluation of chronic headaches (defined as a headache of at least one month's duration) occurring in the absence of any other symptoms or signs suggestive of raised intracranial pressure (ICP). The incidence, causes, management and outcome of chronic headache in these patients was determined. RESULTS: 42 patients were identified, mean age 30 years (range 18 - 59). All had undergone lifelong follow-up. All had previously undergone shunt insertion for hydrocephalus. 16 had undergone endoscopic third ventriculostomy (ETV). 11 had undergone choroid plexus coagulation. 55 % (23/42) of patients underwent investigation for 1 or more episodes of chronic headache. Recurrent hydrocephalus due to shunt malfunction or ETV failure was excluded by ICP monitoring using either an intraparenchymal transducer or monitoring via a ventricular access device. All patients underwent repeat imaging, using CT and/or MR imaging. Identified causes of headache included: shunt blockage; shunt overdrainage; ETV failure and symptomatic Arnold-Chiari malformation. A history of choroid plexus coagulation (CPC) as an infant was associated with a decreased risk of chronic headache in later life (p = 0.02). In 8 patients no definite cause for headaches was identified, in 4 of these patients symptoms resolved spontaneously, the remainder required specialist pain management. CONCLUSIONS: The aetiology of chronic headaches in this patient group is multifactorial. In the absence of other clinical symptoms or signs of raised ICP, ICP monitoring is an invaluable adjunct to management. 10 % of hydrocephalic adult spina bifida patients required specialist pain management for control of chronic idiopathic headache. PMID- 14758562 TI - A study of prenatal ultrasound and postnatal magnetic imaging in the diagnosis of central nervous system abnormalities. AB - Accurate prenatal diagnosis of central nervous system (CNS) abnormalities is essential in counselling parents, as they are the most common developmental abnormalities causing considerable mortality. Currently, the standard in prenatal imaging is ultrasound scanning (USS). The introduction of fast acquisition magnetic resonance imaging (MRI) has lead to increased diagnostic confidence and information available for parents. Frequently USS initially identifies CNS abnormalities as ventriculomegaly alone. However, it is known that ventriculomegaly is commonly associated with other CNS pathology, which may adversely affect the prognosis. As MRI has superior soft tissue resolution and can be used at any time postnatally, it is expected to identify disorders of myelination that may result from prenatal ventriculomegaly. This study will evaluate the role of MRI as a postnatal imaging tool in patients that had a prenatal USS diagnosis of isolated ventriculomegaly. This was a retrospective review of patient notes and scan reports. The postnatal MRI study group consisted of 9 patients that had been diagnosed initially with prenatal isolated ventriculomegaly on USS, and followed up with postnatal MRI (cases of spina bifida and Dandy-Walker malformations were excluded). Findings from the scan reports were recorded and analysed. Both MRI and prenatal USS gave the same information in 55.6 % of the patients. In the remaining 44.4 %, MRI added to the information provided by the prenatal USS. An interesting finding was that MRI missed a small fluid-filled cyst and an arachnoid cyst in 2 cases. 55.6 % of patients went on to develop other CNS abnormalities prenatally, whereas 33.3 % showed prenatal regression of VM with no other pathology. 11.1 % showed postnatal persistence of isolated VM. As USS has the advantage of being cheap and easy to perform, it will remain as the primary imaging tool in obstetric care. MRI can provide significant additional information that can affect parent counselling, prenatal intervention, and postnatal management. Postnatally, MRI can give some idea of prognosis by evaluating myelination patterns, which is not possible with USS. PMID- 14758563 TI - President's Lecture--"A foot in two camps". PMID- 14758564 TI - Good news for our old journal. PMID- 14758565 TI - Hormonal and nutritional regulation of adipose tissue mitochondrial development and function in the newborn. AB - Growth, development, and maturation of adipose tissue in the fetus can determine both survival at birth as well as having longer term consequences for adult disease. The mitochondrial proteins uncoupling protein (UCP) 1, voltage dependent anion channel (VDAC), and cytochrome c have an important role in cellular energy regulation. Activity of these proteins is particularly important during the transition from fetal to neonatal life when cellular energy requirements are at near maximal rates. The regulation of these proteins by endocrine factors is highly complex and may be dependent on both fetal number and maternal nutrition. The cytokine hormones leptin and prolactin have well established functions in energy regulation and lactation respectively. However, recent data proposes a role in regulation of mitochondrial proteins, particularly UCP1, and thermogenesis. Cortisol is an adrenal hormone with a critical role in fetal tissue maturation, especially the lung. It has now been shown to influence the abundance of UCP1 in the fetus, a role that may in part be regulated by the metabolically active thyroid hormone triiodothyronine. A greater understanding of the regulation of mitochondrial proteins within adipose tissue by endocrine and nutritional factors is likely to be important in preventing neonatal morbidity and mortality. It could also add substantially to our understanding of pathological conditions such as obesity and non-insulin dependent diabetes. PMID- 14758566 TI - Expression of leptin and leptin receptor during the development of liver fibrosis and cirrhosis. AB - Leptin is involved in the regulation of food intake and is mainly secreted by adipocytes. Major secretagogues are cytokines such as TNF-alpha or IL-1. Leptin in turn upregulates inflammatory immune responses. Elevated leptin serum levels have been detected in patients with liver cirrhosis, a disease frequently associated with elevated levels of circulating cytokines as well as hypermetabolism and altered body weight. Recently, leptin has been detected in activated hepatic stellate cells in vitro and an involvement of leptin in liver fibrogenisis has been suggested. The current study was designed to further clarify the role of leptin in liver disease by characterizing leptin and leptin receptor expression in the development and onset of experimental liver fibrosis. Liver fibrosis and cirrhosis was induced in rats by use of phenobarbitone and increasing doses of CCl (4). Leptin and leptin receptor mRNA expression was determined by semiquantitative RT-PCR, protein expression by Western blot analysis and localization of leptin and its receptor by immunohistochemistry. Normal liver tissue does not express leptin, but leptin receptor mRNA. Increasing levels of leptin mRNA were detected in fibrotic and cirrhotic livers correlated to the degree of fibrosis. Leptin receptor mRNA expression was not significantly altered in damaged livers. Increasing levels of leptin were detected in fibrotic and cirrhotic livers, whereas protein expression of the receptor remained unchanged. Throughout different stages of liver fibrosis, leptin immunoreactivity was localized in activated hepatic stellate cells only, whereas immunoreactivity for the receptor was mainly seen on hepatocytes. In conclusion, leptin is expressed at increasing levels in activated hepatic stellate cells in vivo, which may therefore be a source of increased leptin tissue and serum levels contributing to the pathophysiology and morphological changes of chronic liver disease. PMID- 14758567 TI - Estradiol stimulates vascular endothelial growth factor and interleukin-6 in human lactotroph and lactosomatotroph pituitary adenomas. AB - Estrogens are considered to be critically involved in lactotroph and lactosomatotroph pituitary tumor development. In addition to direct effects, estradiol-induced tumor formation may involve alterations in growth factor and cytokine production. We have studied whether estradiol stimulates the production of the angiogenic vascular endothelial growth factor and the potential tumor progression factor interleukin-6 in 5 lactotroph (LA) and 5 lactosomatotroph (LSA) human pituitary adenoma cell cultures. All tumors secreted heterogenous basal amounts of VEGF (18.0 +/- 1.4 to 425 +/- 26 pg/ml per 24 h) and IL-6 (18.1 +/- 1.5 to 604 +/- 17 pg/ml per 24 h). Estradiol (100 nM) significantly enhanced VEGF release in all LA and LSA cell cultures (47 to 168 % above basal). IL-6 secretion was stimulated in 3 out of 5 LA and in all LSA cell cultures (31 to 287 % above basal). In cell cultures obtained from tumors from which sufficient cells could be isolated, a dose-dependent effect of estradiol (1 to 100 nM) on VEGF and IL-6 production was observed. Stimulation of IL-6 and/or VEGF secretion by estradiol in the majority of human lactotroph and lactosomatotroph adenoma cell cultures studied, suggests that estrogens may contribute to adenoma expansion through the stimulation of these auto-/paracrine-acting adenoma progression factors. PMID- 14758568 TI - Hormonal profile of men with premature balding. AB - OBJECTIVE: Premature androgenic alopecia has been suggested as a feature of the male equivalent of the syndrome of polycystic ovary. However, the hormonal pattern of men with premature balding has been investigated in only a few studies with inconsistent results. MATERIAL AND METHODS: We examined 37 men with premature balding (defined as frontoparietal and vertex hair loss before the age of 30 years with alopecia defined as grade 3 vertex or more on the alopecia classification scale of Hamilton with Norwood modification). The plasma concentrations of total testosterone, dihydrotestosterone, epitestosterone, androstenedione, cortisol, 17-OH-progesterone (17OHP), estradiol, LH, FSH, prolactin, SHBG and TSH and free thyroxine were measured. RESULTS: The frequency of subnormal values in SHBG, FSH, testosterone and epitestosterone (but not in free androgen index) was significant in the balding men. A borderline significant trend was recorded with respect to increased levels in 17OH-P and prolactin. CONCLUSIONS: The hormonal pattern of a substantial number of men with premature balding resembles in some respects the hormonal pattern of women with polycystic ovary syndrome. PMID- 14758569 TI - Differential target molecules for toxicity induced by streptozotocin and alloxan in pancreatic islets of mice in vitro. AB - Streptozotocin (STZ) and alloxan (ALX) are potent diabetogens in different species of laboratory animals. Here, we describe differential in vitro effects of STZ and ALX on beta-cell molecules that are essential for glucose transport and metabolism, the glucose transporter 2 (GLUT2) and glucokinase (GK), respectively. Incubation of isolated pancreatic islets of C57 BL/6 mice with STZ or ALX for 30 min resulted in a concentration-dependent gradual loss of beta-cell function as determined by basal and D-glucose (D-G)-stimulated insulin release. ALX concentration-dependently reduced the mRNA expression of GLUT2 and GK and the effect on GLUT2 was more marked. STZ, in contrast, did not affect the mRNA expression of GLUT2 and GK, but concentration-dependently reduced the GLUT2 protein expression. Both STZ and ALX failed to affect the mRNA expression of proinsulin and of beta-actin. The deleterious effects of STZ and ALX were not due to beta-cell loss, because the total RNA yields and protein contents as well as the proinsulin mRNA expression in isolated islets of the differentially treated islets did not differ significantly from controls. Furthermore, islets that had been exposed to STZ or ALX responded to the non-glucose secretagogue arginine in a pattern comparable to that of solvent-treated cultures. When preincubating islet cultures with either D-G or its chemically closely related analogue 5-thio D-glucose (5-T-G), different effects were obtained after treatment with either ALX or STZ. Thus, preincubation with 5-T-G protected the cultures from STZ induced GLUT2 protein reduction, whereas D-G failed to do so. Preincubation with D-G, however, protected the cultures from ALX-induced reduction of GLUT2 and GK mRNA expression, whereas 5-T-G, at best, exerted a modest protection against ALX at a concentration of 1 mmol/l. Apparently, in vitro, GLUT2 protein is a key target molecule for STZ, while GLUT2 mRNA and GK mRNA are target molecules for ALX. PMID- 14758570 TI - Concentration of insulin-like growth factor (IGF)-I in iliac crest bone matrix in premenopausal women with idiopathic osteoporosis. AB - Previous studies have shown a link between low serum insulin-like growth factor-I (IGF-I) and decreased bone mass of patients with osteoporosis. However, whether serum levels are representative for the growth factor concentration or activity available in human bone tissue is controversial. In the present study, IGF-I was assessed in serum and bone matrix extracts from the iliac crest in 19 eugonadal women with idiopathic osteoporosis and in 38 age-matched controls. In addition, the relationship between the skeletal levels of IGF-I and bone mineral density (BMD) or the susceptibility to osteoporotic fractures in women with osteoporosis was examined. Bone matrix extraction was performed based on a guanidine HCL/ethylendiamine-tetraacetic acid (EDTA) method. No significant difference in both serum and bone matrix IGF-I levels between groups was observed. Serum IGF-I concentrations failed to be associated with bone matrix IGF-I levels in osteoporotic patients. However, in premenopausal women with idiopathic osteoporosis, skeletal IGF-I positively correlated with BMD at the lumbar spine (r = + 0.58, p = 0.01). In contrast, neither femoral neck BMD nor Ward's triangle BMD was associated with bone matrix IGF-I concentrations. A tendency towards lower levels of bone matrix IGF-I in subjects with vertebral fractures as compared to those without fractures was observed in age-adjusted analyses, however the difference failed to remain statistically significant after adjustment for bone mineral density. These data provide no clear evidence for low bone matrix IGF-I as a determinant factor of age-unrelated osteoporosis. However, low skeletal IGF-I concentrations may aggravate osteoporosis in these women. PMID- 14758571 TI - Plasma homocysteine after insulin infusion in type II diabetic patients with and without methionine intolerance. AB - BACKGROUND: A high prevalence of hyperhomocysteinemia has been reported in type II diabetic patients with documented vascular disease; hence the hypothesis that hyperhomocysteinemia may contribute to overall mortality in diabetic patients. The link between insulin and homocysteine metabolism has not been completely clarified yet; in particular, only few data are available on the effects of insulin in vivo on homocysteine metabolism in the presence of abnormalities of sulphur amino acid metabolism (methionine intolerance). MATERIALS AND METHODS: To establish whether methionine intolerance and which of its determinants could influence total plasma homocysteine in response to insulin infusion in vivo in type II diabetic patients, we submitted 18 patients (Group A) with normal and 18 patients with abnormal (hyperhomocysteinemia) (Group B) response to oral methionine load to a glucose/clamp study. At time 0, and 30, 60 and 120 minutes after hyperinsulinemia, homocysteine and methionine plasma levels were assessed. In order to evaluate the cause of methionine intolerance, all patients were assayed for fasting homocysteine-cysteine ratio (as a marker of suspected heterozygosis for cystathionine-beta-synthase deficit), MTHFR C (677)T status and homocysteine-related vitamin status (serum vitamin B (6) [PLP], vitamin B (12) and folate). RESULTS: After hyperinsulinemia, plasma methionine was reduced (by about - 30 % at 120 minutes vs. basal values) within both groups, whereas tHcy tend to decrease in group A following insulin administration (up to - 6.6 +/- 3.6 % vs. basal values at 120 minutes) with a significantly higher variability, while in patients with "methionine intolerance" (group B) tHcy tended to increase (up to + 29.05 +/- 8.3 % vs. basal values at 120 min from the clamp). Serum folic acid (7.45 +/- 2.8 vs. 4.82 +/- 2.5 nmol/L, p < 0.05), Vit. B (12) (348 +/- 78 vs. 242 +/- 65 pmol/L, p < 0.05) and PLP (84.1 +/- 23.6 vs. 50.6 +/- 32.4 nmol/L; p < 0.01) were significantly higher in group A than in group B; PLP levels significantly correlated with homocysteine after 4 h methionine load (n = 36; r = - 0.327, p < 0.05); group A showed also a significantly lower prevalence of suspected heterozygosis for cystathionine-beta-synthase deficit (1/18 [11.1 %] vs. 5/18 [33.3 %], p < 0.05) and MTHFR T allele presence (4/18 [22.2 %] vs. 11/18 [61.1 %], p < 0.01). A stepwise regression analysis with tHcy plasma level variations (event A = reduction; event B = increase) as the dependent variable showed that low serum folate and PLP levels and presence of MTHFR T allele were the variables associated with insulin-induced tHcy increase. CONCLUSIONS: Methionine intolerance may influence the effect of insulin administration on plasma homocysteine in patients affected by type 2 diabetes. To prevent a possible acute (and repeated) hyperhomocysteinemia due to insulin administration in cases of methionine intolerance, it may be useful to assess the presence of methionine intolerance (tHcy after oral methionine loading) and Hcy-related vitamin status in all patients due to be subjected to insulin therapy. PMID- 14758572 TI - Calcitonin measurement to detect medullary thyroid carcinoma in nodular goiter: German evidence-based consensus recommendation. AB - Serum calcitonin (CT) has become a very specific and sensitive marker for human medullary thyroid carcinoma (MTC), a neuroendocrine tumor affecting about 1 % of patients with nodular thyroid disease. MTC is characterized by early micrometastasis and a lack of curative non-surgical treatment, so that early diagnosis is desirable. Based on a systematic review of scientific evidence, we propose multidisciplinary consensus recommendations for the clinical use of CT in patients with nodular goiter. To exclude MTC, serum CT should be determined in patients with nodular thyroid disease, using a two-site CT immunoassay. If basal serum CT exceeds 10 pg/ml, CT should be analysed by pentagastrin stimulation testing, after renal insufficiency and proton pump inhibitor medication have been ruled out. As the risk for MTC is higher than 50 % in patients with stimulated CT values > 100 pg/ml, thyroidectomy is advised in these individuals. If stimulated CT exceeds 200 pg/ml, thyroidectomy and lymphadenectomy is strongly recommended. Pentagastrin-stimulated CT values < 100 pg/ml are associated with a low risk of MTC, or very rarely, non-metastasizing micro-MTC (size < 10 mm). Therefore, regular clinical and biochemical follow-up is the preferred treatment in such patients, unless thyroid malignancy is suspected otherwise. PMID- 14758573 TI - No major effect of orciprenaline and propranolol upon ACTH-induced cortisol secretion. AB - Preclinical research suggests adrenal beta-adrenergic receptors to be involved in the regulation of steroid synthesis. In a group of healthy male volunteers, we compared ACTH-induced cortisol and dehydroepiandrosterone (DHEA) secretion after pre-treatment with orciprenaline, propranolol or placebo. Neither baseline nor ACTH-induced steroid secretion differed between these conditions. Our data do not support the hypothesis that the adrenal beta-receptor plays a major role in steroid secretion in humans. PMID- 14758574 TI - A case of Kallmann syndrome associated with Dandy-Walker malformation. AB - A 19-year-old man was admitted to our hospital for delayed puberty. At birth, he had macrocephalia and showed delayed physical and mental development. At 9 years of age, right cryptorchism was diagnosed. His parents had noticed that he could not recognize any smells since his infancy. Physical examination on admission revealed ocular hypertelorism, high myopia, high arched palate, and intermittent external strabismus. Sense of smell was scaled out by olfactometry. External genitalia were infantile. Neurological examination showed on IQ of 83, and mild truncal ataxia. Magnetic resonance imaging (MRI) showed a cystic distension of the IV ventricle, partial aplasia of the cerebellar vermis, elevation of the tentorium cerebelli, enlargement of the III ventricle, and agenesis of the corpus callosum. These findings revealed that the patient had Dandy-Walker malformation. The basal FSH, LH, and testosterone levels were all low compared with normal adult reference values. The serial LH-RH provocation tests showed stepwise LH and FSH elevation. After the fifth day of LH-RH administration, both LH and FSH responses clearly improved. Olfactory tracts were defective in MRI findings. These findings were consistent with hypogonadotropic hypogonadism of hypothalamic origin with anosmia, and the patient was therefore diagnosed with Kallmann syndrome. Sequence analysis of the KAL1 gene showed no mutation in the coding region. To our knowledge, this is the first case report of the coexistence of Kallmann syndrome and Dandy-Walker malformation in the same patient. PMID- 14758575 TI - Identifying undiagnosed dementia in residential care veterans: comparing telemedicine to in-person clinical examination. AB - BACKGROUND: Dementia is a common but frequently undiagnosed problem in aging. Barriers to early diagnosis include a lack of routine screening for dementia and a lack of access to specialty consultative services. We conducted a pilot study to see if telemedicine could provide reliable, accurate geriatric consultative services to evaluate patients for dementia who were residing at remote sites. METHODS: This was a prospective cohort study that compared the diagnostic reliability of telemedicine to an in-person examination for dementia. Eligible subjects were residents of a Washington State Veterans' Home, age 60 years or older, with no prior diagnosis of dementia. Eligible subjects were screened for dementia using the 7-Minute Screen. Veterans who screened positive and consented to participate in the study received an in-person neuropsychiatric evaluation at baseline, and then both telemedicine and in-person examinations for dementia conducted by experienced geriatric psychiatrists. The accuracy of the telemedicine diagnosis was estimated by comparing it to the diagnosis from the clinical examination. Three geriatric psychiatrists who were blinded to the results of the clinical examination conducted the telemedicine and in-person examinations. We also assessed attitudes of the subjects and geriatric psychiatrists towards the telemedicine sessions. RESULTS: Eighteen of 85 subjects screened were 'positive' for dementia on the 7 Minute Screen. Of these, 16 consented to participate in the telemedicine study. Twelve of the 16 subjects were subsequently diagnosed with dementia by the telemedicine examination. The telemedicine diagnoses were in 100% agreement with the diagnoses from the in person clinical examinations. Moreover, the subjects reported a high degree of satisfaction with the telemedicine experience and that they would like to have further care through telemedicine in the future. The geriatric psychiatrists reported technical difficulties with the audio-visual quality of telemedicine in the initial phases of the project that resolved as familiarity with the telemedicine equipment increased. None of these problems had an adverse impact on the diagnostic accuracy of telemedicine. CONCLUSIONS: We found that telemedicine was as accurate as an in-person clinical examination in establishing the diagnosis of dementia. In addition, subjects reported a high degree of satisfaction with telemedicine and a willingness to participate in telemedicine clinical care in the future. Given the large increase in the aging population and the shortage of geriatric psychiatrists nationally, it appears that telemedicine may be a promising means to expand the availability of geriatric psychiatric consultation to remote areas. PMID- 14758576 TI - Caudate volume measurement in older adults with bipolar disorder. AB - BACKGROUND: Decreased caudate volumes have been noted in unipolar depressed subjects, especially in the elderly and those with cognitive impairment. No differences have been noted in initial studies of multi-aged bipolar subjects; however, this region has not been examined in older bipolar subjects. METHODS: We examined the caudate nuclei volumes of 36 older bipolar subjects (mean age 58) and 35 older controls (mean age 62) using logistic regression analyses to control for age and gender differences. Differences between late- and early-onset (age-of onset before age 45) bipolar subjects were also examined, as well as the effect of length of illness. RESULTS: The right caudate was noted to be smaller in older bipolar subjects compared with older controls when controlled for sex and age (p = 0.0448). No differences were noted in overall brain volume nor lateral ventricular volume between the bipolar and control subjects. Late-onset bipolar subjects had a decrease in brain volume (p = 0.035) compared with early-onset bipolar subjects. Late-onset bipolar subjects had a decrease in the right (p = 0.044) and total (p = 0.04) caudate size compared with older controls. CONCLUSIONS: Right caudate volume is decreased in older bipolar subjects compared to controls. Bipolar subjects with late-onset illness have significantly decreased right and total caudate volumes compared to controls. This is affected by neither the length of illness nor the age of onset. Late-onset bipolar subjects have decreased total brain volume compared with early-onset bipolar subjects. PMID- 14758577 TI - Olanzapine versus placebo in the treatment of psychosis with or without associated behavioral disturbances in patients with Alzheimer's disease. AB - OBJECTIVES: Psychotic symptoms and behavioral disturbances are a concern in the care of elderly patients with Alzheimer's dementia (AD). This study was conducted to compare the efficacy of olanzapine versus placebo in patients with psychotic symptoms associated with AD in long-term or continuing-care settings. METHODS: Patients (n = 652) with AD and delusions or hallucinations were randomly assigned to 10 weeks of double-blind treatment with placebo or fixed-dose olanzapine (1.0, 2.5, 5.0, 7.5 mg/day). RESULTS: Mean age was 76.6+/-10.4 years. Repeated-measures analysis showed significant improvement from baseline in NPI/NH Psychosis Total scores (sum of Delusions, Hallucinations items-primary efficacy measure) in all five treatment groups (p<0.001), but no pairwise treatment differences were seen at the 10-week endpoint. However, under LOCF analysis, improvement in the 7.5 mg olanzapine group (-6.2 +/- 4.9) was significantly greater than with placebo (-5.0 +/- 6.1, p = 0.008), while endpoint CGI-C scores showed the greatest improvement in the Olz 2.5 olanzapine group (2.8 +/- 1.4, p = 0.030) relative to placebo (3.2 +/- 1.4). There were significant overall treatment-group differences in increased weight, anorexia, and urinary incontinence, with olanzapine showing numerically higher incidences. However, neither the incidence of any other individual events, including extrapyramidal symptoms, nor of total adverse events occurred with significantly higher frequency in any olanzapine group relative to placebo. No clinically relevant significant changes were seen across groups in cognition or any other vital sign or laboratory measure, including glucose, triglyceride, and cholesterol. CONCLUSIONS: While 1.0 mg olanzapine did not show significant differences from placebo, the 2.5 mg dose was a reasonable starting dose. Olanzapine at 7.5 mg/day significantly decreased psychosis and overall behavioral disturbances (NPI/NH, BPRS) and was well tolerated. PMID- 14758578 TI - Reasons of informal caregivers for institutionalizing dementia patients previously living at home: the Pixel study. AB - CONTEXT: Study of the problems and requirements of the main caregiver providing home care for dementia patients that have resulted in the patient being institutionalised. OBJECTIVES: To determine the reasons for placing the dementia patient in an institution. RESOURCES: Self-administered questionnaire of 48 questions on the patient and caregiver, including a list of complaints, given to the main caregiver. Medical questionnaire on the patient filled in by the geriatrician. RESULTS: Data were collected from 109 questionnaires concerning 75 females with dementia (84.7 +/- 6.7 years) and 34 demented males (80.8 +/- 7.4 years). In two-thirds of cases the main caregiver was a female, aged 61.1 +/- 12.1 years. Cognitive disorders were not the main reasons for institutionalizing patients. The most frequent caregiver complaint at the time of institutionalisation was incontinence, followed by withdrawal. The caregiver's main problem resulting in institutionalisation was dependence, with behavioural disorders in second place. A treatment with anticholinesterase for dementia was associated with a live-in career being provided for 20 months longer than in the case of patients not receiving this treatment. Statistical analysis revealed 6 groups of separate caregiver-patient situations. On the one hand there were those patients who appeared to be easy for the caregiver to cope with: those with no problems, docile patients and passive patients not opposing care. In these cases the caregiver was most often young and male, or not directly related to the patient. On the other hand there were 3 other groups: patients with inappropriate motor behaviours, violent/agitated patients and unmotivated patients who opposed care. These patients lived with an elderly caregiver who had been looking after the patient for several years. DISCUSSION: Caregivers' requirements are for help with coping with and preventing dependence. The caregiver suffers terribly from a lack of relief, particularly when young. CONCLUSION: It is necessary to change the focus of home care for dementia patients towards preventing loss of autonomy and its consequences and to allow for periods of relief for home caregivers. PMID- 14758579 TI - DemTect: a new, sensitive cognitive screening test to support the diagnosis of mild cognitive impairment and early dementia. AB - OBJECTIVES: To design a new, highly sensitive psychometric screening to identify patients with mild cognitive impairment (MCI) and patients with dementia in the early stages of the disease. METHODS: Five tasks were included in the DemTect: a word list, a number transcoding task, a word fluency task, digit span reverse, and delayed recall of the word list. The normation was performed with 145 healthy control subjects (CG). Furthermore, 97 MCI patients and 121 patients with possible Alzheimer's disease (AD) were tested with the DemTect and the MMSE. Classification rates for both tests were analysed. RESULTS: On the basis of the CG data, age-dependant transformation algorithms for the DemTect subtests were defined, and an education correction was provided for the total transformed score. The patient groups scored significantly below the CG in both the DemTect and the MMSE. Compared to the MMSE, classification rates of the DemTect were superior for both the MCI and the AD group, with high sensitivities of 80% and 100%, respectively. CONCLUSIONS: The DemTect is short (8-10 minutes), easy to administer, and its transformed total score (maximum 18) is independent of age and education. The DemTect helps in deciding whether cognitive performance is adequate for age (13-18 points), or whether MCI (9-12 points) or dementia (8 points or below) should be suspected. PMID- 14758580 TI - Patient predictors of response to treatment of depression in Alzheimer's disease: the DIADS study. AB - OBJECTIVE: To investigate patient predictors of response to treatment of Major Depressive Episode (MDE) in Alzheimer's disease (AD). METHODS: Forty-four outpatients with AD and MDE were randomized to receive either sertraline or placebo in a 12-week placebo-controlled, flexible-dose clinical trial after a one week single-blind placebo phase. All participants were evaluated for depression at entry using the 21-item Hamilton Depression Rating Scale (HDRS) and the Cornell Scale for Depression in Dementia (CSDD). All subjects completed baseline neuropsychological testing. Caregiver burden and depression were also measured. The forty-two subjects who completed at least one post-enrollment follow-up visit were included in the analysis. RESULTS: No baseline demographic, mood, neuropsychiatric, neuropsychological, or caregiver variable was a statistically significant predictor of response to treatment. There were trends for African American patients (p=0.07) and those with milder baseline agitation/aggression (p=0.08) to respond better. CONCLUSION: No baseline characteristic assessed clearly predicts response to treatment of MDE in AD. A diverse population of depressed AD patients may thus respond similarly to the same treatment. PMID- 14758581 TI - Disclosing a diagnosis of dementia: a systematic review. AB - BACKGROUND: The issue of diagnostic disclosure in dementia has been debated extensively in professional journals, but empirical data concerning disclosure in dementia has not previously been systematically reviewed. OBJECTIVE: To review empirical data regarding diagnostic disclosure in dementia. METHODS: Five electronic databases were searched up to September 2003 (Medline, Embase, Cinahl, Sociological Abstracts, Web of Science). Additional references were identified through hand searches of selected journals and bibliographies of relevant articles and books. The title and abstract of each identified paper were reviewed independently by two reviewers against pre-determined inclusion criteria: original data about disclosure were presented and the paper was in English. Any disagreements were resolved by discussion until consensus was reached. Data were extracted independently by two reviewers using a structured abstraction form. Data quality were not formally assessed although each study was critically reviewed in terms of methodology, sampling criteria, response rates and appropriateness of analysis. RESULTS: Fifty-nine papers met the inclusion criteria for detailed review. Many of the studies had methodological shortcomings. The studies reported wide variability in all areas of beliefs and attitudes to diagnostic disclosure and reported practice. Studies of the impact of disclosure indicate both negative and positive consequences of diagnostic disclosure for people with dementia and their carers. CONCLUSIONS: Existing evidence regarding diagnostic disclosure in dementia is both inconsistent and limited with the perspectives of people with dementia being largely neglected. This state of knowledge seems at variance with current guidance about disclosure. PMID- 14758582 TI - Care arrangements for people with dementia in developing countries. AB - BACKGROUND: Rapid demographic ageing will soon lead to large increases in the numbers of persons with dementia in developing countries. This study is the first comprehensive assessment of care arrangements for people with dementia in those regions. METHODS: A descriptive and comparative study of dementia care; caregiver characteristics, the nature of care provided, and the practical, psychological (Zarit Burden Interview, General Health Questionnaire) and economic impact upon the caregiver in 24 centres in India, China and South East Asia, Latin America and the Caribbean and Africa. RESULTS: We interviewed 706 persons with dementia, and their caregivers. Most caregivers were women, living with the person with dementia in extended family households. One-quarter to one-half of households included a child. Larger households were associated with lower caregiver strain, where the caregiver was co-resident. However, despite the traditional apparatus of family care, levels of caregiver strain were at least as high as in the developed world. Many had cutback on work to care and faced the additional expense of paid carers and health services. Families from the poorest countries were particularly likely to have used expensive private medical services, and to be spending more than 10% of the per capita GNP on health care. CONCLUSIONS: Older people in developing countries are indivisible from their younger family members. The high levels of family strain identified in this study feed into the cycle of disadvantage and should thus be a concern for policymakers in the developing world. PMID- 14758583 TI - Alzheimer Disease International's 10/66 Dementia Research Group - one model for action research in developing countries. AB - BACKGROUND: The 10/66 Dementia Research Group (10/66) founded in 1998, is a network of over 100 researchers from mainly developing countries. 10/66 is committed to encourage more good quality research in those regions, where an estimated two-thirds of all those with dementia live. It represents a collaboration of academics, clinicians, and an international non-governmental organization, Alzheimer's Disease International (ADI). METHOD: 10/66 pilot studies in 26 centres in Latin America, India, Africa and China and SE Asia suggest that education and culture-fair diagnosis is an attainable aim. Despite extended family care networks, these studies also identified high levels of practical, psychological and economic strain upon caregivers. Population-based studies in six centres will now estimate prevalence, describe impact and seek to identify genetic and environmental risk factors in novel settings. At a practical level, 10/66 has studied ways to circumvent the lack of help-seeking in developing countries, and has developed a low-level intervention to educate and train caregivers. CONCLUSION: The links with ADI and its international networks, and the volunteerism of ADIs members have fostered the rapid growth of 10/66. The partnership facilitates both the raising of awareness and influence upon policy, as 10/66 research evidence can be used by ADI and national Alzheimer's Associations to direct and support advocacy. PMID- 14758584 TI - The impact associated with caring for a person with dementia: a report from the 10/66 Dementia Research Group's Indian network. PMID- 14758585 TI - Occurrence and treatment of post-traumatic stress disorder in an elderly patient after a traffic accident. PMID- 14758586 TI - Comparison study of venlafaxine and paroxetine for the treatment of depression in elderly Chinese inpatients. PMID- 14758587 TI - The burden of providing care for Alzheimer's disease patients in Poland. PMID- 14758588 TI - Population growth impairment of aliphatic alcohols to Tetrahymena. AB - The toxicity of a series of 120 aliphatic alcohols was evaluated using the Tetrahymena pyriformis population growth impairment assay. For tertiary propargylic alcohols; primary, secondary, and tertiary homopropargylic alcohols; allylic alcohols; and saturated alcohols, a statistically robust structure activity model was developed for toxicity data [log (IGC(50) (-1))] using the 1 octanol/water partition coefficient (log K(ow)) as the lone descriptor [log (IGC(50))(-1) = 0.74 (log K(ow)) - 1.73; n = 97; r(2) (adj.) = 0.933; r(2) (pred.) = 0.932; s = 0.298; F = 1328; Pr > F = 0.0001]. Analysis of data for the primary propargylic alcohols yielded a separate, high-quality log K(ow)-dependent quantitative structure-activity relationship (QSAR) [log (IGC(50))(-1) = 0.65 (log K(ow)) - 1.22; n = 10; r(2) (adj.) = 0.969; r(2) (pred.) = 0.964; s = 0.222; F = 254; Pr > F = 0.0001]. A comparison of the observed toxicity and that predicted by the first QSAR showed that the primary propargylic alcohols with log K(ow) values < 2.00 exhibited enhanced toxicity and that this increased toxicity was inversely related to hydrophobicity. In sharp contrast, analysis of the data for the secondary propargylic alcohols exhibited little relationship with log K(ow) (r(2) = 0.339). Although the initial QSAR can be used to model the toxicity of any aliphatic alcohol for the T. pyriformis population growth impairment end point, the estimated potency would be underestimated for primary propargylic alcohols with log K(ow) values < 2.00. Moreover, estimates of toxic potency of secondary propargylic alcohols based on this QSAR should be viewed with limited confidence. The findings for beta-unsaturated alcohols in Tetrahymena were sharply different from that reported for fathead minnow acute mortality; this difference in toxicity is a result of a difference in the protocol used rather than in metabolism. PMID- 14758589 TI - Ultrastructural alterations in the liver and kidney of white sea bass, Lates calcarifer, in acute and subchronic cadmium exposure. AB - Ultrastructural alterations in the liver and kidney of 3-month-old white sea bass, Latescalcarifer, after cadmium exposure were studied by transmission electron microscopy (TEM). One group of fish was exposed to a cadmium concentration of 10 mg/L (acute) for 96 h in a static system, and another group was exposed to cadmium concentrations of 0.8 and 3 mg/L cadmium (subchronic) for 3 months in a recirculation closed system. Ultrastructural alterations observed in the hepatocytes included mitochondrial condensation, swelling, and lysis. The rough endoplasmic reticulum (RER) showed dilation, fragmentation, and vesiculation. After subchronic exposure there were numerous large lipid droplets and abundant stored glycogen. Ultrastructural alterations observed in the proximal tubules of the kidney included nuclear degeneration, condensation, and massive swelling of the mitochondria; RER fragmentation and vesiculation. Disorganized brush borders and increased numbers of large hydropic vacuoles and lysosomes were also observed. PMID- 14758590 TI - Characteristics of microcystin production in the cell cycle of Microcystis viridis. AB - The correlation between the content of three microcystins (types LR, RR and YR) and the cell cycle of an axenic strain of Microcystis viridis, NIES-102, was investigated under conditions of high (16 mg L(-1)) and low (1.0 mg L(-1)) nitrate (NO(3)-N) concentrations. Each phase of the cell cycle was identified using a flow cytometer equipped with a 488-nm argon laser using SYTOX Green dye, which binds specifically to nucleic acids and can be exited by the wavelength (Ex/Em: 504/523 nm on DNA). Microcystin concentration showed a positive linear correlation with DNA concentration. The microcystin content of the cells changed remarkably as the cell cycle process proceeded, with maximum content in the G(2)/M phase and minimum content in the G(0)/G(1) phase. Under a condition of high NO(3)-N concentration, the ratio of the total content in the G(0)/G(1) phase to that in the G(2)/M phase was about 6:1. In contrast, for the two batch cultures the total content was 1.3-fold greater in the G(2)/M phase. The compositions of the three microcystins also changed along with the cell cycle process, although there was little difference in composition that was related to NO(3)-N concentration. Therefore, there were distinctive compositions specific to each phase of the cycle, and the cell cycle of the M. viridis strain was more strongly responsible for both the quantity and the types of microcystin production than was the effect of NO(3)-N concentration. PMID- 14758591 TI - Toxicity of arsenic species to Lemna gibba L. and the influence of phosphate on arsenic bioavailability. AB - The toxicity of arsenic (As) species to Lemna gibba L. and the influence of PO(4) (3-) on As bioavailability and uptake were tested in batch culture. L. gibba were exposed to six test concentrations of NaHAsO(4). 7H(2)O and NaAsO(3), with 0, 0.0136, 13.6, and 40 mg L(-1) KH(2)PO(4). In batch culture As toxicity to L. gibba did not relate linearly to As concentration. The growth rate, related to frond number as recommended by OECD and ISO/DIN, was significantly inhibited in fronds exposed to 20-50 microg L(-1) As(III) compared with fronds exposed to As(V). The growth rate was stimulated when plants were exposed to 50-250 microg L(-1) of both As(III) and As(V). After exposure to 300-800 microg L(-1) growth inhibition was significantly higher for As(III) than for As(V), whereas above 800 microg L(-1) As(V) was inhibited the most. The bioaccumulation of As(III) and As(V) was significantly higher for P-deficient cultures (0.98 +/- 0.08 and 1.02 +/- 0.19 g kg(-1), respectively for 0.0136 mg L(-1) PO(4) (3-)) than for P sufficient cultures (243 and 343 mg kg(-1) for 40 mg L(-1), respectively). Plants exposed to As(V) had uptake and accumulation values slightly higher than did plants exposed to As(III). No significant differences in bioaccumulation were found between plants exposed to a concentration of As(III) >1 mg L(-1) and those exposed to As(V) at the same concentration. This indicates a direct relationship to P content in the culture. Toxicity may result from the uptake of As(V) instead of PO(4) (3-) as a result of ion competition during uptake because of close thermodynamic properties, which may change the interaction among components in the media. The toxicity pattern is interpreted as a manifestation of changing speciation in the batch culture and of the oxidation of As(III) to As(V) in an oxygen-rich environment. PMID- 14758592 TI - Mercury concentrations in migratory waterfowl harvested from Southern Nevada Wildlife Management Areas, USA. AB - Mercury concentrations were determined in 14 species of migratory waterfowl harvested from Southern Nevada Wildlife Management areas during the 2001-2002 hunting season. Common mergansers (2.61 +/- 0.87 ppm liver; 0.22 +/- 0.04 ppm muscle), northern shovelers (3.51 +/- 3.8 ppm liver; 0.16 +/- 0.03 ppm muscle), and bufflehead (2.63 +/- 0.24 ppm liver; 0.91 ppm muscle) had the highest concentrations of mercury in the liver and muscle of the species harvested. The relationships between muscle and liver concentrations were also examined. These data indicate that liver tissue can be used with reasonable confidence (r >.80 for most species) to predict mercury concentrations in muscle, the most commonly consumed portion of waterfowl. The mercury concentrations reported here are some of the highest reported in the scientific literature, and they identify certain species, such as the northern shoveler and common merganser, that have accumulated unusually high concentrations of mercury. Evidence for the use of these three species as possible bioindicators for mercury is also presented. PMID- 14758593 TI - Aquatic toxicity of four alkylphenols (3-tert-butylphenol, 2-isopropylphenol, 3 isopropylphenol, and 4-isopropylphenol) and their binary mixtures to microbes, invertebrates, and fish. AB - The acute and chronic toxicity of four simple alkylphenols with butyl and propyl substitutions was evaluated with aquatic microbes, invertebrates, and fish. These alkylphenols-3-tert-butylphenol, 2-isopropylphenol, 3-isopropylphenol, and 4 isopropylphenol-have been detected in various environmental media, but their impact on aquatic fauna has seldom been evaluated. Relative susceptibility to each phenolic varied by test species. The marine bacterium Vibrio fischeri was the most susceptible to the alkylphenols, up to 3 orders of magnitude more sensitive than species of higher trophic levels. For 4-isopropylphenol, the 5-min Microtox EC(50) value was 0.01 mg/L, whereas the EC(50) for Ceriodaphnia after a 48-h exposure was 10.1 mg/L. Notable differences in sensitivity to the alkylphenols was also observed with the Microtox assay: 4-isopropylphenol was > 200 times more toxic to V. fischeri than was 2-isopropylphenol (EC(50) = 2.72 mg/L). For V. fischeri, the mixture toxicity of the alkylphenols was additive in nature and was predicted by a concentration addition model. The energy of the lowest unoccupied molecular orbital (ELUMO) explained the observed toxicity of the individual alkylphenols to V. fischeri (r(2) = 0.92, p < 0.05). These results suggest that the mode of action of polar narcotic alkylphenols to V. fischeri is different than that of other test organisms, possibly because of the differences in the cell structure of the prokaryotic V. fischeri. PMID- 14758594 TI - Evaluation of flushing of a high-selenium backwater channel in the Colorado River. AB - Concern has been raised that selenium contamination may be adversely affecting endangered fish in the upper Colorado River basin. The objective of the study was to determine if operation of a water control structure (opened in December 1996) that allowed the Colorado River to flow through a channel area at Walter Walker State Wildlife Area (WWSWA) would reduce selenium and other inorganic elements in water, sediment, aquatic invertebrates, and forage fish. Endangered Colorado pikeminnow were collected and muscle plug samples taken for selenium analysis. Selenium concentrations in filtered water were 21.0 microg/L in 1995, 23.5 microg/L in 1996, 2.1 microg/L in 1997, and 2.1 microg/L in 1998. Selenium concentrations in sediment cores and sediment traps were 8.5 microg/g in 1995, 8.2 microg/g in 1996, 4.8 microg/g in 1997, and 1.1 microg/g in 1998. Selenium concentrations in aquatic invertebrates were 27.4 microg/g in 1996, 15.5 microg/g in 1997, and 4.9 microg/g in 1998. Selenium concentrations in forage fish were 27.2 microg/g in 1996, 20.2 microg/g in 1997, and 8.6 microg/g in 1998. Selenium concentrations in muscle plugs of Colorado pikeminnow were 9.8 microg/g in 1995, 9.5 microg/g in 1996, 9.0 microg/g in 1997, and 10.3 microg/g in 1998. Although selenium concentrations in water, sediment, aquatic invertebrates, and forage fish decreased substantially after operation of the water control structure, a corresponding change in Colorado pikeminnow did not seem to occur. Selenium concentrations in muscle plugs decreased with increasing fish total length and weight, did not change between repeat sampling in the same year or recapture in subsequent years, and seemed to be most closely associated with the mean monthly river flow for the March-July period. PMID- 14758595 TI - Extraction of cyanobacterial endotoxin. AB - To simplify our efforts in acquiring toxicological information on endotoxins produced by cyanobacteria, a method development study was undertaken to identify relatively hazard-free and efficient procedures for their extraction. One article sourced and two novel methods were evaluated for their ability to extract lipopolysaccharides (LPSs) or endotoxins from cyanobacteria. The Limulus polyphemus amoebocyte lysate (LAL) assay was employed to compare the performance of a novel method utilizing a 1-butanol-water (HBW) solvent system to that of Westphal's (1965) phenol-water system (HPW) for the extraction of endotoxin from various cyanobacteria. The traditional HPW method extracted from 3- to 12-fold more endotoxin from six different cyanobacterial blooms and culture materials than did the novel HBW method. In direct contrast, the novel HBW method extracted ninefold more endotoxin from a non-microcystin producing Microcystis aeruginosa culture as compared to the HPW method. A solvent system utilizing N,N' dimethylformamide-water (HDW) was compared to both the HPW and HBW methods for the extraction of endotoxin from natural samples of Anabaena circinalis, Microcystis flos-aquae, and a 1:1 mixture of Microcystis aeruginosa/Microcystisflos-aquae. The LAL activities of these extracts showed that the novel HDW method extracted two- and threefold more endotoxin from the Anabaena sample that did the HBW and HPW methods, respectively. The HDW method also extracted approximately 1.5-fold more endotoxin from the Microcystis flos aquae sample as compared to both the HBW and HPW methods. On the other hand, the HBW method extracted 2- and 14-fold more endotoxin from the Microcystis flos aquae/Microcystis aeruginosa mixture than did the HPW and HDW methods, respectively. Results of this study demonstrate that significant disparities exist between the physicochemical properties of the cell wall constituents not only of different cyanobacterial species but also of different strains of the same cyanobacterial species, as showing by the varying effectiveness of the solvent systems investigated. Therefore, a sole method cannot be regarded as universal and superior for the extraction of endotoxins from cyanobacteria. Nevertheless, the ability of the novel HBW and HDW methods to utilize easily handled organic solvents that are less hazardous than phenol render them attractive alternatives to the standard HPW method. PMID- 14758596 TI - Metallothionein induction in aquatic oligochaete tubifex tubifex exposed to herbicide isoproturon. AB - Metallothioneins (MTs) are low-molecular-weight proteins mainly involved in metal ion detoxification. Recently it has been demonstrated that MTs participate in several cellular functions such as regulation of growth and antioxidative defenses. Moreover, pesticides can induce their synthesis. The aim of the current work was to determine the effects of isoproturon, either pure or formulated as Matin (suspension containing an isoproturon concentration of 500 g. L(-1)), on the metallothionein and total protein contents of the aquatic worm Tubifex tubifex. MT levels in exposed worms increased significantly after 7 and 15 days of exposure to a concentration of the herbicide of 50 mg. L(-1). Isoproturon reduced the metal (Cu, Zn, and Cd) content of metallothioneins, and it also increased the total protein content of the worms. These results suggest that MT induction may not be considered a specific biomarker of metal exposure but that it can be used as a nonspecific biomarker of the effect of isoproturon effect in aquatic worms. PMID- 14758597 TI - Direct nucleic acid diagnostic tests for bacterial infectious diseases: streptococcal pharyngitis, pulmonary tuberculosis, vaginitis, chlamydial and gonococcal infections. AB - Partnering with a reference lab can actually help grow a referring lab's own business. The reference lab provides a hospital's physicians with access to unique tests and clinical information and broadens the scope and depth of the referring lab's menu. As medicine moves toward prescribed treatments based on a patient's individual genotype or phenotype, the reference lab will play an even bigger role in helping the hospital to educate physicians and their patients to the most efficient testing strategies consistent with good patient management. PMID- 14758598 TI - The role of esoteric reference laboratory as a partner for growth. AB - Partnering with a reference lab can actually help grow a referring lab's own business. The reference lab provides a hospital's physicians with access to unique tests and clinical information and broadens the scope and depth of the referring lab's menu. As medicine moves toward prescribed treatments based on a patient's individual genotype or phenotype, the reference lab will play an even bigger role in helping the hospital to educate physicians and their patients to the most efficient testing strategies consistent with good patient management. PMID- 14758599 TI - Abbott Diagnostics' Joe Nemmers concentrates on trend-setting issues. Interview by Dottie Dunham. PMID- 14758600 TI - The impact of marker selection and process improvements on cardiac care. PMID- 14758601 TI - The standardization of troponin I assays: an update. PMID- 14758602 TI - Automated slide stainers for SS, IHC, and ISH: A review of current technologies and commercially available systems. PMID- 14758603 TI - Addressing management issues. Dealing with a controlling boss. PMID- 14758604 TI - Laboratory access: when to withdraw the "welcome mat". PMID- 14758605 TI - Congress challenged to resolve severe medical lab personnel shortage. PMID- 14758606 TI - The quality of family planning programs: concepts, measurements, interventions, and effects. AB - This study reviews the major research and interventions concerning readiness and quality of care in family planning programs. It has three aims: to identify and describe the principal methodological research including conceptual frameworks, perspectives, and tools for measuring and improving quality; to describe the results from various intervention studies; and to assess what is known about the effect of such interventions. The review suggests that interventions that improve client-provider interactions show the greatest promise. Good quality of care results in such positive outcomes as clients' satisfaction, increased knowledge, and more effective and longer use of contraceptives. Rigorously documented evidence of the effects of interventions is sorely needed. The review indicates areas requiring additional research. PMID- 14758607 TI - Adolescents in Vietnam: looking beyond reproductive health. AB - The research that has been conducted to date on Vietnamese adolescents has focused on unprotected and unsanctioned sexual activity and its health consequences, specifically abortion and sexually transmitted diseases, especially HIV. The question we pose in this article is whether this concern is warranted. Is the population community justified in limiting research on this population to early sexual activity and HIV risk? Even if the sexual behavior of young people can be considered problematic, are there perhaps other aspects of young peoples' lives to which more attention should be devoted? The literature on adolescent sexual behavior in Vietnam is reviewed and data on premarital sex and reproductive behavior are analyzed from a 1999 survey conducted in six provinces among nearly 1,500 adolescent boys and girls aged 15-22. Descriptive data on schooling and work are included in order to put the information on sexual activity in perspective. The data analysis reveals that, at least currently, the sexual behavior of unmarried adolescents in Vietnam is not what jeopardizes their health and well-being. PMID- 14758608 TI - Determinants of contraceptive method choice in rural Tanzania between 1991 and 1999. AB - Four pooled Demographic and Health Survey data sets are used to examine the determinants of contraceptive method choice in rural Tanzania for the period from 1991 to 1999. The individual data are linked to facility surveys conducted in the same communities so that the impact of Tanzania's family planning program can be examined. The focus of the study is an examination of the effect on method choice of the three major components of Tanzania's family planning program: logistical support, trained providers, and communications programs. The statistical methods employed correct for the potential endogeneity of family planning message recall. Simulations are used to quantify the impact of the important policy variables. All three components of the program are shown to have had an impact on modern method choice. PMID- 14758609 TI - Abortion service provision in South Africa three years after liberalization of the law. AB - In 1996, South Africa introduced legislation that liberalized women's access to termination of pregnancy. This study presents the findings of a survey undertaken to describe the availability and accessibility of abortion services in 1999, three years after the law was passed. All facilities that had been officially designated to perform these services were contacted by telephone to determine whether they were providing the services, their capacity, whether they were performing second-trimester as well as first-trimester terminations, and how long women had to wait for these services. Nationally, 292 facilities had been designated, but in 1999 only 32 percent were functioning. Of the functioning facilities, 27 percent were in the private sector. Mapping of available services indicated that substantial parts of the country were entirely without such services. Half of the country's induced abortions were being performed in Gauteng province, although only 19 percent of women of reproductive age were living there. This finding indicates that service provision in other provinces was inadequate or lacking. Although in the first years following the new legislation efforts were made to establish abortion services, this study reveals gross inequality in service availability. Strategies for improving coverage are suggested. PMID- 14758610 TI - Malawi 2000: results from the Demographic and Health Survey. PMID- 14758611 TI - Turkey 1998: results from the Demographic and Health Survey. PMID- 14758612 TI - [Evoked potentials in the brain of adolescents in norm and those with attention deficit during recognition of short-term acoustic stimuli]. AB - The effect of stimulus duration on auditory event-related potentials and performance of oddball task was studied in normal children and those with attention-deficit symptoms. Mismatch negativity was absent on presentation of short-term (11 ms) stimuli and present with longer stimuli (50 ms). The adolescents with deficit of attention performed much worse (errors of omission) with the short stimuli. The RT was significantly larger in subjects with attention-deficit with all types of tested stimulus duration. They also manifested a smaller P3b amplitude in response to task-relevant deviant stimuli and larger N2b peaks in response to the standard stimuli. It was possible to differentiate between the MMN and the N2b components owing to the fact that the MMN was absent with shorter stimuli. The findings suggest that there is a deficit in processing of sensory information at the cortical level in subjects with the attention-deficit symptoms. PMID- 14758613 TI - [Manifestations of selective visual attention in human parietal and temporal cortex according to evoked potentials]. AB - The event-related potentials (ERPs) in visual discrimination task in parietal and temporal cortical areas were recorded in 11 young adults during passive observation (involuntary attention) and target selection (voluntary attention). The voluntary selective attention resulted in: 1) increased ERP correlation between the parietal; and temporal cortical areas; 2) increased correlation of sequential ERPs in monopolar leads (P3, P4, T3, T4, T5, T6); and 3) increased correlation of sequential ERPs in bipolar leads (P3-T3, P3-T5, P4-T4, P4-T6). The findings suggest that voluntary attention maintains a concordant activity of the parietal and temporal cortical areas in execution of visual selection tasks. PMID- 14758614 TI - [Effect of impulse extrabroad-band electromagnetic radiation on electroencephalogram and sleep in laboratory animals]. AB - 1-hour exposure to ultra-short impulse low-frequency (6 Hz) superbroad band electromagnetic radiation altered cortical EEG in rats just after the exposure and increased the paradoxical sleep in rabbits within 16-22 hours following the radiation. PMID- 14758615 TI - [Hypothalamic monoamines in the cold stress during changes in activity of nitric oxide system]. AB - Effects of NO-synthase inhibitor N(omega)-nitro-L-arginine (LNA) and donor sodium nitroprusside (SNP) on alteration in body temperature, plasma corticosterone level and hypothalamic monoamines in response to cold exposure, were studied. Drop of the body temperature in cold exposure in rats treated with LNA or SNP was the same as in the control group. Administration of SNP (2 mg/kg i.p.) significantly increased the basal level of corticosterone (CS). Cold exposure elevated CS in all groups of rats. LNA did not markedly alter the hypothalamic noradrenaline (NA) while SNP significantly decreased the NA. Cold exposure resulted in additional decrease of the NA in SNP-treated rats. NA was found to significantly increase within 48 hrs following the cold exposure in the LNA as well as in the SNP groups. SNP significantly increased basal dopamine and DOPAC levels. Cold exposure did not affect hypothalamic dopamine. In the experiments, NO changes of serotonin and 5-hydroxyindoleacetic acid were observed. The findings suggest that antagonistic effects of the NO-synthase inhibitor and NO donor postulated in literature for various kinds of stress do not occur in experiments with cold stress. PMID- 14758616 TI - [Cardio-electric field during heart ventricle repolarization in hens]. AB - Recovery sequence of ventricular epicardium and body surface potential distribution during the ST-T complex in birds, were studied. Recovery sequence of ventricular epicardium in hens appeared to be independent of activation sequence and to be governed by activation-recovery intervals. Inversion of mutual location of positive and negative areas on the body surface in hens corresponded to the end of the QRS complex and the beginning of the ST-T complex. PMID- 14758617 TI - [Endothelium-dependent mechanism of formation of the systemic hemodynamics responses]. AB - In anaesthetised rats, effects of blockade of the NO-synthetase upon hemodynamic shifts were studied (arterial pressure, cardiac output, general peripheral vascular resistance), the shifts being evoked either by increase (infusion of polyglucon) or by decrease (orthostasis) in the cardiac output. Under the blockade of the NO-synthetase, the pressor effects of polyglucon increased by 27% and the orthostatic hypotension by 72%. Responses of general peripheral vascular resistance changed in the same direction. The findings suggest importance of the NO secretion by the vessels' endothelium for formation of the systemic hemodynamics responses. PMID- 14758618 TI - [Application of elliptical cylinder as a novel model of microvessel shape for analysis of thyroid capillaries in hypercalcemia]. AB - Previously designed stereological method for estimating axial ratios (X0/Y0) of microvessels (MVs) whose shape is approximated by elliptical cylinders was applied for analysis in the rat thyroid perifollicular hemocapillaries in hypercalcemia (Ca gluconate--10% i.m., 0.7 ml/day, for 3 days). Electron microscopy revealed the size of major (X) and minor (Y) radii of ectioning profiles of the capillaries. The X/Y distribution corresponded to use of characteristics foe elliptical cylinder model's stochastically geometric 3D/2D relation of distribution of two--or three-dimensional (3D), and observed, or two dimensional (2D), axial ratio values. The X0/Y0 values of the MVs under study were estimated as X0/Y0 approximately 1.3 for 79% and X0/Y0 approximately 2.7 for 21% of capillaries. The estimates obtained are of comparable value with the X0/Y0 calculated previously for the thyroid capillaries of the normal rat. They can be employed in physiological studies of the MVs. PMID- 14758619 TI - [Age-related changes of lipid spectrum, level of lipid peroxidation, and antioxidant defence in the liver and blood of rats]. AB - It has been shown that the lowest level of total lipids, cholesterol, triacylglycerols and products of lipid peroxidation of blood and liver, as a rule, is specific to adult rats. These characteristics are significantly higher for old and young animals. At the same time, the level of glutathione and alphatocopherols in adults' liver is much higher than in young and old rats. It suggests the lower level of processes of lipid peroxidation in adult mature rats. The relative high level of products of lipid peroxidation and low content of alpha-tocopherols in old rats' liver (against the background of higher activity and glutamineperoxidase, and glutathionereductase than in adults) suggests tocopherol deficiency in old animals. High content of total lipids, cholesterol and cholesterol entering into the composition of lipoprotein of different density, triacylglycerols, diene conjugates and malonic dialdehide, activity of glutathione-dependent enzymes of antioxidant defence in young animals as compared with these levels in adult rats seems to be associated with agerelated hypercholesterolemia and intensive plastic changes of a growing organism. PMID- 14758620 TI - [Effect of C-terminal peptide of G-protein alfa(s)-subunit on regulation of adenylate cyclase and protein kinase A activities by biogenic amines and glucagon in mollusc and rat muscles]. AB - The peptide synthesised by us: 387-394-amide (10(-7)-10(-4) M), in a dose dependent manner decreases activities of adenyl cyclase and proteinkinase A evoked by serotonine and glucagon in smooth muscles of the freshwater bivalve mollusc Anodonta cygnea and that evoked by beta-agonist isoproterenol--in the rat skeletal muscles. Even in concentration 10(-7) M, the peptide completely eliminates potentiation of the hormones' stimulating effect on adenylyl cyclase activity with the non-hydrolizable analogue of guanine nucleotides (Gpp[NH]p). At the same time, the peptide does not affect stimulation of the adenylyl cyclase activity with non-hormonal agents (NaF, Gpp[NH]p and forkolin). In the presence of the peptide, inhibiting effects of the hormones on activities of adenylyl cyclase and protein kinase A will be preserved. The findings reveal the importance of the G-protein alpha s-subunit's C-terminal regional of a stimulating type for its functional coupling with receptors of a serpentine type, and elucidate the molecular mechanisms of interaction between the G-protein and the receptor. PMID- 14758621 TI - [Function of the hypothalamo-pituitary-adrenal system in mice with ectopic overexpression of agouti protein]. AB - Agouti protein (AP) is known to antagonise the effects of melanocortins (ACTH, MSH) on the melanocortin receptors which participate in regulation of central and peripheral HPA links. This study aimed at estimation of effects of dominant mutation Agouti yellow (Ay-ectopic overexpression Agouti protein) on the HPA axis function in mice. Male mice of C57B1/6J strain of Ay/a- and a/a genotypes (control animals, lack of AP), were used. We demonstrated that basal corticosterone level in the Ay mice matched those of control animals. Stress activated corticosterone level (p < 0.02) and sensitivity of adrenal to low doses ACTH in vitro and in vivo were higher in Ay mice compared with control mice. Dexamethazone-inhibition of stress-reactivity was more intensive in Ay/a than in a/a mice (p < 0.0007). PMID- 14758622 TI - [Anxiety in females caused by long-term psycho-emotional stress]. AB - Behaviour of female mice was studied under long-lasting psychoemotional influence. Daily presence of females during inter-male confrontations through a perforated transparent partition in the same cage and the next female placing on the male territory (sawdust) after the confrontations, were included as the influencing factors. Under these conditions, the increased level of anxiety in the females was revealed in the elevated plus-maze test. At the same time, the females demonstrated a more passive behaviour in the Porsol test. The psycho emotional influence also affected the females' behaviour in the partition test. The females spent a significantly longer time near the partition both 5 minutes prior to and during the inter-male confrontations. PMID- 14758623 TI - [Psycho-emotional manifestations in rats with hippocampal lesions]. AB - The study aimed at revealing psycho-emotional manifestations of Wistar rats in a problem situation when they had to solve a food-getting task in a multi alternative maze. Bilateral lesion of dorsal hippocampus did not affect the learning process while the pattern of psycho-emotional manifestations changed in all the animals irrespective of their individual properties. The pverall effect was manifested in flatness of psycho-emotional responses as a result of reduction of extreme forms of both passive and active stress responses. Individual effects depended on the behavioural phenotype. Hippocampus lesion in initially excitable rats (10%) resulted in reduction of active stress responses (act/pass = 0.35/0.5 instead of 1.0/0.4 in control). Hippocampus lesion in initially inhibited rats (30%) did not affect the basic pattern of psycho-emotional responses, while in the rats with initially depressed cognitive activity (60%) it resulted in decreasing of passive and increasing of active stress responses (act/pass = 0.45/1/4 instead of 0.3/1.9). The findings suggest that hippocampus takes part in the psycho-emotional responses while the individual psycho-emotional pattern is determined by the morpho-functional features responsible for organisation of a psycho-emotional condition. PMID- 14758624 TI - [Effect of natural zeolites on renal functions and water-salt metabolism in rats]. AB - In vivo experiments on adult Wistar rats, it has been found that intake of natural zeolites resulted in temporary decrease of renal water, sodium and potassium excretion. At the same time, reabsorption of water and electrolytes increased. This effect was due to the stimulation of Na+, K(+)-ATPase activity in thick ascending limb of Henle loop and hormonal changes: increase of insulin, thyroxin and aldosterone concentration in plasma. The water and ion content in most of the tissues under study was higher in the experimental group than in control. It has been suggested that renal response in rats with zeolites intake was compensatory lower as a result of gastrointestinal losses of ions and ion accumulation in tissues. PMID- 14758625 TI - [Thermoregulation in the comfort temperature zone]. AB - Under constant adequate temperature conditions (comfortable for a man) the retention of temperature homeostasis was shown to require a continuous functioning of the physiological thermoregulation system to prevent short-term and whole day deviations of the temperature from the physiological level. Under adequate temperature conditions the thermoregulation system was shown to attain its highest sensitivity and accuracy. It is possible that this occurs owing to physiological control over the total body heat content being included into the process of thermoregulation. The data are given on the existence and "structure" of the physiological mechanisms of such a control. PMID- 14758626 TI - [Pregnancy and lactation suppress development of genetic obesity in female mice with Agouti yellow mutation]. PMID- 14758627 TI - [Disorders of cognitive processes in simulation of Alzheimer's disease in monkeys]. AB - Two groups of monkeys were learned to differentiate stimuli with different types of information and to make a spatial choice. Characteristics of the operative memory were revealed in the delayed differentiation tasks prior to and after administration of p75-saporin (I group) and saline (II group). For the first time the Alzheimer disease in monkeys was shown to entail a deficit of operative memory due to disorders in the sensory and cognitive components of the memory. The degree of reduction of the correct decision making was shown to depend on the delay duration and the type of visual information. Following the saline administration, no significant changes occurred in the monkeys (II group). The data obtained suggest that structural-functional organisation of the cholinergic and noradrenergic mechanisms predetermining the sensory processing, differs from those involved in decision-making. PMID- 14758628 TI - [Features of stepping pattern formation in decerebrated cats under epidural spinal cord stimulation]. AB - The mechanisms of stepping pattern formation initiated by epidural spinal cord stimulation in decerebrated cats, were investigated. It is shown that the ability to produce the stepping pattern involve the L3-L5 segments. In flexor muscle, the formation of stepping pattern under optimal stimulation frequency (5-10 Hz) of these segments is provided by polysynaptic activity with the latency 80-110 ms. In extensor muscle, this process is realized through interaction of monosynaptic reflex and polysynaptic activity. The stepping pattern under epidural stimulation is determined by spinal structures with modulation influence of the peripheral feedback. PMID- 14758629 TI - [Control of voluntary movements mediated via propriospinal neurons]. AB - Cortico- and rubrospinal tracts play an important role in controlling voluntary movements. Transection of these tracts in different spinal cord layers gives different effects that may be explained by the influence of different spinal cord neuronal networks. The aim of the present work was to study the role of C3/C4 propriospinal system in movement control and processes of motor recovery. It was shown that propriospinal system C3/C4 play crucial role in motor recovery after lesion of cortico- and rubrospinal tracts in C5, whereas ventrally located tracts are important after the same lesion in C2. More over, propriospinal system C3/C4 can mediate the command for some voluntary movements in cats. PMID- 14758630 TI - [Effect of 6-hydroxydopamine on the electrical characteristics of snail neurons in long-term sensitization]. AB - Studies of the influence of neurotoxin 6-hydroxydopamine selectively destroying the catecholamine terminals on long-term sensitization, and the role of dopamine in manifestation of characteristics of a membrane of identified neurons during elaboration of plasticity, were fulfilled. Injection of saline was used as the control. It is shown that preliminary injection of 6-hydroxydopamine reduces duration of long-term sensitization, but does not block it completely. It was shown that injection of 6-hydroxydopamine prevents diminishing of membrane and threshold potentials in withdrawal interneurons during formation of long-term sensitization. The experiments demonstrate that shift of electrical characteristics of withdrawal interneurons caused by injection of neurotoxin 6 hydroxydopamine to both naive snails and sensitized snails, statys during at least 10 days. PMID- 14758631 TI - [Effects of L-dopa and transcranial magnetic stimulation on behavioral reactions in kindled rats]. AB - In acute experiment in rats, the chronic epileptogenesis was reproduced in the form of pharmacological kindling induced via repeated picrotoxin administrations (1.0-1.2 mg/kg, intraperitoneally). A reduction of exploratory behavior was shown in the early period of kindling (24 h as of the moment of the last epileptogen administration). The marked alleviation of these disturbances was registered in two weeks from the moment of cessation of kindled irritations. L-DOPA (100 mg/kg, intraperitoneally) and transcranial magnetic stimulation (20 impulses with an induction at the height of their development of 1.5 TI) was followed by the net increasing exploratory, sexual and eating behavior. This is in favour of regarding the activation of dopaminergic system as a mechanism of action of transcranial magnetic stimulation upon kindling-induced behavioral deterioration. PMID- 14758632 TI - [Activity of nucleolar organizers in central macrophage culture of bone marrow erythroblastic islands]. AB - The erythroblastic islands of the bone marrow are morphofunctional units of erythropoiesis. In this work the functional state of erythroblastic islands' cells of the bone marrow, for the first time, was defined by the estimation of the activity of the nucleolar organizers of central macrophages in the erythroblastic islands, cultivated during 24 and 48 hours with the presence of various doses of erythropoietin. The findings indicated that the increase in doses of erythropoietin was accompanied by the corresponding increase of the activity of nucleolar organizers in central macrophages of erythroblastic islands. The nucleolar organizers of central macrophages in cultures of erythroblastic islands responded to very small doses of erythropoietin by their activation. PMID- 14758633 TI - [Features of active control used in pharmacological studies]. AB - The effects of 5 injections of salt solution and unfractionary heparin in dose 0.36 microgram/kg (Serva, Germany 10 kDa, activity 180 U/mg) have been studied in Wistar rats. It was found that two injections of salt solution were enough to form a stable defensive state in all rats which was manifested as an expectation of pain in tail-flick testing. The defensive motivation provoked by the injections negatively influenced the learning process as saline-induced rats refused to solve a food-getting task in a problem situation. Explorative and locomotor activities were depressed in these rats and were accompanied by numerous stressful and neurotic-like manifestations. Unlike saline-rats, practically all heparinized-rats instead of 45% of intact rats were able to solve a cognitive task despite the injections. Anxiety was decreased, but sensitivity to different external factors was increased in the heparin-induced rats. Formed habit in these rats was characterized by a high organization and stability. However, the majority heparin effects in tail-flick test were discovered when comparing the heparin-induced rats with intact ones and were not observed in comparison with the saline-rats. It is suggested that the saline-control should be considered as specific control having defensive features which are necessary to take into account in interpretation of effects of other pharmacological preparations. PMID- 14758634 TI - [Alterations of inotropic function of isolated heart and extent of injury of cardiomyocytes after cannabinoid receptor activation during ischemia and reperfusion]. AB - It was found that CB1- and CB2-receptor activation by intravenous administration of the selective CB-agonist HU-210 at a dose 0.1 mg/kg prompts an increase of myocardial resistance to the pathogenic action of ischemia and reperfusion in vitro. The revealed effects of HU-210 do not depend on the activation of CB receptors in the myocardium. PMID- 14758635 TI - [Effect of prolactin on the level of corticosterone in the blood and synthesis of lymphocyte-activating factors by macrophages under conditions of glucocorticoid loading]. AB - The present study examines in vivo the interaction between glucocorticoids and prolactin, and immunomodulating activity of prolactin, including prolactin induced changes in production of lymphocyte-activating factors and alteration in humoral immune response. It was shown that prolactin application increased the level of corticosterone in the rat blood. Administration of prolactin prior to hydrocortisone administration induced alteration in the level of corticosterone that depended on the dose of hydrocortisone. Application of hydrocortisone reduced humoral immune response and lymphocyte-activating factors production by peritoneal macrophages. Administration of prolactin prior to hydrocortisone administration prevents inhibitory action of glucocorticoids/on humoral immune response and lymphocyte-activating factors production by macrophages. PMID- 14758636 TI - [Effect of prenatal stress upon parameters of the behavioral response induced by a tonic pain focus in male and female rats during postnatal ontogenesis]. AB - The long-term effects of prenatal stress (immobilization of females during the last week of pregnancy) were studied on pain sensitivity to a prolonged irritant in Wistar rats during prepubertal, pubertal periods of development and in adults. Pain sensitivity was evaluated from indices of the biphasic behavioral response in the classical formalin test--the intensity of patterns of flexes, shakes and time spent licking, and from duration of the first, acute and the second, tonic phases and from the duration of interphase. Consequences of the prenatal stress manifested themselves differently in the patterns of the response organized at the spinal and supraspinal levels, during tonic phase mainly, differently in females and males; there were peculiarities of effects of prenatal stress in each age period. The data obtained suggest that the consequences of the prenatal stress at the spinal level manifest themselves in activation of modulating descending facilitating and in suppression of inhibitory monoaminergic systems in prepubertal and pubertal rats, and on the contrary, in activation of descending inhibitory and suppression of facilitating systems in adults. Furthermore, considerable evidence is obtained corroborating the idea of inhibitory nature of the interphase and the mechanisms of modulation of acute and tonic phases in the formalin test during different age periods of the individual development. PMID- 14758638 TI - [Stimulation of thermoregulatory and respiratory functions with the help of EDTA and EGTA during deep hypothermia in rats]. AB - Influence of EDTA (C10H14N2Na2O8.2H2O) and EGTA (C14H24N2O10) on physiological functions homoiothermic organisms at deep hypothermia, was studied. White rats during cooling were in special sections without rigid fixing of head and limbs. In reply to intravenous introduction of EDTA and EGTA solutions, similar answers of the organisms were observed: raised breathing frequency and amplitude, intensity of electrical activity of muscles; these signs of activation of physiological functions lasted 8-10 minutes. Besides, of the 20th-30th minute after introduction of the second dose of preparations (at rectal temperature 17.1 +/- 0.5 degrees C), the secondary activation respiratory and thermoregulatory functions were registered. The termination of the cold shivering in experiments with introduction of EDTA and EGTA solutions occurred at lower temperatures in rectum and in a brain (16.7-17.3 degrees and 17.8-18.2 degrees C, resp.) than in control experiments (18.7 +/- 0.6 degrees C and 20.2 +/- 1.5 degrees C). The authors suppose that the activation of the thermoregulatory and respiratory functions is caused by a decrease in concentration of ions Ca2+ in the blood plasma. PMID- 14758637 TI - [Deficiency of glucocorticoid production delays the healing of acute gastric mucosal erosion and chronic ulcers in rats]. AB - Effects of glucocorticoid deficiency followed by corticosterone replacement on the healing of gastric erosions and chronic gastric ulcers have been investigated in rats. Glucocorticoid deficiency was induced by adrenalectomy performed after the formation of gastric erosions or ulcers. Gastric erosions were produced by indomethacin (35 mg/kg, i.p.) or by 6 h immobilization at temperature 8 degrees C, chronic gastric ulcers were induced by 60% acetic acid. All ulcerogenic stimuli caused an increase in corticosterone production. Adrenalectomy created corticosterone deficiency and delayed the healing of gastric erosions and chronic gastric ulcers. The effect of adrenalectomy was more evident in the indomethacin ulcerogenic model. Replacement by corticosterone prompted the healing of gastric erosions and ulcers in adrenalectomized animals. These data suggest a participation of endogenous glucocorticoids in a restoration of gastric mucosal integrity. PMID- 14758639 TI - [Functional features of actinin and tropomyosin distribution in rat kidney]. AB - The intrarenal distribution of actinin and tropomyosin was studied by western blot analysis in various functional conditions. It was found that actinin content is always higher in cortex than in medulla. The highest tropomyosin content was revealed in outer medulla, but it is more than twice higher in rats of mutant Brattleboro line versus hydrated normal WAG rats. This ratio rises up to 46.12 +/ 2.14 versus 13.83 +/- 0.66 (in relative units) for rats being under dehydration that maximally activated vasopressin secretion in normal WAG rats. PMID- 14758640 TI - [Progesterone transport across the wall of the rumen of sheep]. AB - Data on content of fiber, lignin and progesterone in reticulo-ruminal chyme and progesterone in blood serum and saliva of sheep throughout the reproductive cycle and at intramuscular injection of progesterone, are presented. A direct correlation between concentration of progesterone in blood and chyme was revealed. 1.5% of progesterone entered the reticulo-rumen with saliva. Transport of hormone through the wall of forestomach is the main way of progesterone entering the reticulo-rumen. The transport is carried out against the gradient of hormone concentration. Possible participation of progesterone adsorption on lignin in explanation of this transport, is discussed. PMID- 14758641 TI - [Effect of substance P, neurokinin A and a selective agonist of tachykinin receptors of the NK-2 type on bile metabolism]. AB - Based on the experiments on rats, changes of the level of bile secretion and chemical composition of bile following application of substance P, neurokinin A, and selective agonist of the NK-2 tachykinins receptors neurokinin A Fragment (4 10), are interpreted. These results demonstrate that tachykinins (SP, NKA, and NKA Fragment (4-10)) are true holeretics and affect various enzyme systems in the hepatocyte. The alterations of the coefficients of bile confirm that. PMID- 14758642 TI - [Recognition of visual images by separate hemispheres in conditions of masked blocking of interhemispheric transmission]. AB - The subjects learned to recognize three figures presented in the left visual hemifield and three figures presented in the right visual hemifield. During presentation of a stimulus, the contralateral hemifield was overlapped by a mask. After the training, recognition of all six figures presented in the right and left visual hemifields, was compared. Each hemisphere recognizes figures which were learned in the corresponding visual hemifield, but the recognition of figures learned in the opposite visual hemifield was poor. Thus, the ability of the hemispheres to act separately in recognizing different sets of visual images, was established. PMID- 14758643 TI - [Contrast detection thresholds during their presentation in right and left hemifields of vision]. AB - We measured contrast detection thresholds for thin black vertical bars (1 min of arc wide, 1 deg long) against a bright background, which were presented eccentrically at 4 deg to the left or right of fixation and flanked on one side by a subthreshold bar having the same dimensions. On each presentation, the stimulus randomly appeared in the left or right visual hemifield. The separation between the test and the flanks varied from 1 to 60-120 min of arc. Different subjects showed better performance in different hemifields. This preference was lost or changed to opposite in the course of training. The central excitatory areas are more or less symmetrical, but inhibitory areas reveal asymmetrical tendency: when flanked bars were more peripheral than the test line, the inhibitory areas were larger. Our results show absence of hemispheric specificity in the line detection task. We discuss the ways of stimulus description in fovea and periphery. PMID- 14758644 TI - [Efficiency of a visual system under conditions of uncertainty of the appearance of a moving object]. AB - The effect of uncertainty of a moving object appearance in the noise field upon the coefficient efficiency, we studied. At short durations of presentation (40-80 msec) and high level of external noise, this effect was maximal: magnified 100 times. The efficiency coefficient dependence on the duration of a moving object presentation was shown to be characterized by two maximums. The position of minimum situated between these two maximums was found to be independent of either presence or absence of uncertainty of a number of parameters: such as initial position of the object the image, time of its appearance, noise level, velocity and direction of the movement, and has a latency approximately 120 sec. A functional model of the observed phenomena, has been proposed. PMID- 14758645 TI - [Apparent motion effect under undetermined trajectory of presentation of test dot stimuli]. AB - The influence of such factor as determinacy of trajectory of stroboscopically presented test spot on apparent movement illusion appearance, was studied. Six subjects took part in psycho-physiological experiments during which a test spot was presented successively along the straight line to observer on a display, randomly deviating from this line up or down by 0.39, 0.78, 1.17 or 1.56 angular minutes. It was computed that with the test spot deviating by 0.92 angular minutes from the straight trajectory prognosticated by the observer, the probability of disappearance of apparent straight uniform motion of the spot was equal to 0.75. The findings suggest than one of the conditions under which apparent movement illusion appears involves an agreement between the shape of trajectory of test object presentation as expected by the observer, and its real shape in the experiment. PMID- 14758646 TI - [Dynamics of tuning to the shape of a cross-like figure in striate neurons]. AB - Dynamics of tuning to the shape of cross-like figure flashed in receptive field was studied in 83 striate neurons by the method of temporal slices. Tuning was estimated by the total number of spikes in the response and by this number in successive fragments of the response with 20 ms steps. It was found that only in 11.7% cases neurons showed stable tuning to the same shape of the preferred figure (an angle between its lines), in other cases (88.3%) during response generation this tuning changes being one-phase (7.2%) or two-phase (27.0%), or undulatory (54.1%). Different dynamical reorganization of receptive field zones is discussed as a possible mechanism of the revealed effects as well as their correlation with previously described dynamics of tuning to orientation of a single bar and a cross in striate cells. PMID- 14758647 TI - [Interaction between sensory and cognitive processes in visual recognition: the role of the associative areas of the cerebral cortex]. AB - Monkeys were taught to differentiate stimuli with different types of information and to perform a spatial choice. Development of Alzheimer's disease in experimental monkeys entailed a deficit of operative memory, a considerable enhancement of entropy related to correct decisions. In control monkeys, no significant changes of these characteristics occurred. In monkeys, following a bilateral removal of the sulcus principalis, the 7th field, or both, the operative memory deficit was also determined by two components. The specifics of disorders in the operative memory due to Alzheimer's disease suggest that cholinergic mechanisms determining the sensory processing differ from those involved in decision-making. The structural-functional organization of interaction between sensory and cognitive processes controlled by the motivation and attention system, is discussed. PMID- 14758648 TI - [Temporal summation and reaction time for grid detection]. AB - We studied the stimulus-duration effect on the reaction time (RT) to gratings with different spatial frequencies. At each duration value, the product of the contrast and the duration, i.e. the contrast "energy" was kept constant. We found that at near-threshold "energy" levels the RT was constant for up to 15 ms at lower spatial frequencies and up to 30 ms at higher spatial frequencies. At higher "energy" levels, this critical interval was the same (up to 15 ms) for both lower and higher spatial frequencies. The effect of the duration of gradual onset of the stimulus on RT was also studied. An increase of onset duration (up to 60 ms) at low spatial frequencies substantially delayed reaction time for the contrasts under study. Conversely, at high spatial frequencies, this effect was present only for gratings with a high contrast. These results suggest that reaction time is determined by two types of mechanisms (transient and sustained) at a near threshold contrast, and by one type (transient) mechanism at higher contrasts. PMID- 14758649 TI - [Invariants of spatial summation for S (short wavelength) cone vision]. AB - Spatial summation in the human visual system was studied as a function of retinal eccentricity upon selective stimulation of the short-wavelength sensitive cones. The area of complete spatial summation (Ricco's area) was found to increase with retinal eccentricity while the threshold of stimuli equal in size with Ricco's area remained constant. Comparisons with known morphology of the small bistratified retinal ganglion cells, the only cells known to be excited by S-one ON stimulation, showed that Ricco's area included 2-4 such cells and is up to 1.5 times larger than the dendritic field of a single cell. These relationships were relatively constant within the eccentricity range tested (5-20 deg along the temporal horizontal meridian) and might be the source of threshold invariance of stimuli matching Ricco's area. PMID- 14758650 TI - [Effect of striate visual cortex topography on the parameters of image filtering]. AB - A model of spatial-frequency filtering at the level of 4C layer of the striate visual cortex is proposed and based on the well-known literature data. The evidence on conformable character of representation of the visual field on the primary visual cortex and the suggestions concerning uniformity of horizontal connections of cortical neurones serve as the ground of the model. A correspondence between predictions of the model and results of the experiments with shape perception and size discrimination has been obtained. PMID- 14758651 TI - [Distortions of length perception: anisotropy of the visual field and geometric illusions]. AB - A combined influence of stimulus orientation and structure on the judgement of length was tested in psychophysical experiments. The subjects adjusted the test part of a stimulus to be equal in length to the reference part. The V-shaped stimuli (three dots, or the Oppel-Kundt figure, or one dot and two Muller-Layer wings) were generated on the monitor. In the Oppel-Kundt and Muller-Layer figures, the filled part was considered as a reference and the empty part as a test. In session of the experiments, values of errors measured as functions of orientation of the parts of the stimuli. We assume that experiments with the three-dot stimuli yielded pure characteristics of visual field anisotropy, while those with the Oppel-Kundt and Muller-Layer figures showed a combined effect of both anisotropy and illusions. The data demonstrated that illusions and anisotropy are to be interpreted as independent factors, which converge to an algebraic summation in a simultaneous manifestation. PMID- 14758652 TI - [Cortical neuronal connections and reconstruction of visual space]. AB - We investigated distribution of retrograde-labelled cells in cortical areas 17, 18, and the transition zone 17/18 of both hemispheres in cats after microiontophoretic horseradish peroxidase (HRP) injections into the single cortical columns of area 17, 18, 19 or 21a. On the base of clustered pattern of intrinsic labelling, asymmetric location of labelled callosal cells that was associated with the appropriate pattern of labelling in layers A and A1 of lateral geniculate nucleus, we suggest that cortical neuronal connections are eye specific and may provide for each eye a separate binding of visual hemifields. After HRP injections into columns of area 19 or 21a, the disparate inputs from areas 17, 18 and transition zone 17/18 were revealed. Such connections may provide a local depth information and the selection of stereoscopic surfaces in central sectors of visual space. PMID- 14758653 TI - [Electrical activity of the retina in different axial lengths of the eye in humans]. AB - In subjects with normal vision, we studied the relationship between the axial length of the eye and the amplitude-temporal characteristics of different types of ganzfield ERG. The amplitude of the ERG had the highest variability in the middle of the range of the axial eye lengths. The variability decreased at both ends of the range. With increasing axial length, the amplitude of the b-wave of all types of ERG decreased, and the latency decreased concurrently. The b-wave reflects the activity of the retinal ON-neurones depolarizing in response to the light stimulus. We assume that the decrease in the amplitude of b-wave may be related to the decreased number of photoreceptors and of neurons in the following retinal levels and/or increased inhibition in proximal retina, as well as an increase in relative activity of retinal OFF-neurones hyperpolarizing in response to the light stimulus. PMID- 14758654 TI - [Problem of visual segmentation and spatial-frequency filtration]. AB - The analysis of the visual segmentation problem involved questions concerned with a signal description as a result of linear filtering, nature following nonlinear transformations, and possible role of a spatial attention in pooling. Different approaches to the visual segmentation problem are compared. Our data suggest a model of visual segmentation. It uses the universal principle of linear filters pooling at second-order mechanisms. The pooling mechanism is characterized by position, spatial frequency and orientation tuning an is selective in respect to pattern size. The most activated pooling mechanisms are "targets" for selective attention. It is shown that the information selected this way is necessary and sufficient for recognition. Multiple representation of a visual scene with different resolution and hierarchical principle of description organise the recognition as successive specification. PMID- 14758655 TI - [Changes of the metabolic activity pattern in neurons of the striate cortex of the cat reared in conditions of flickering illumination]. PMID- 14758656 TI - [Methodical means for investigating visual perception of fragmented images]. PMID- 14758657 TI - [Projections of the gastrointestinal tract organs on afferent ganglions of the vagus nerve in rats]. AB - Experiments were carried in 42 mature white male rats. We investigated into the projections of different organs of gastrointestinal tract on afferent neurones of ganglia of the vagus nerve in white rats. Horseradish peroxidase-conjugated lectins were used as a tracer. We investigated into metric parameters (diameter of an equivalent circle) and shape parameters (circular factor of the shape) of marked neurocytes using a method of computer video-analysis. To evaluate reliability of the received data, methods of non-parametric statistics were used. It was determined that the largest neurocytes in afferent ganglions are involved in innervation of the root of the long and ileocecal angle, the smallest ones--in innervation of a cervical department of esophagus and liver. The viscero- and somatosensory neurocytes in caudal ganglion of vagus nerve involved in innervation of different organs in white rat, are characterized by selectivity of the shape and by metric parameters. PMID- 14758658 TI - [Evaluation of EEG(D2) and CNV correlation dimension in various forms of visual attention in human]. AB - In 16 healthy subjects, the EEG and the CNV (contingent negative variation) correlation dimension was studied during preparing to pay attention to one of three attention tasks: the situations "What", "Where" and "What and where". Two easily discernible symbols (squares and circles) were used as stimuli. The findings show that, irrespective of the attention form, the correlation dimension of the EEG, the CNV amplitude, and the evoked potential's N1-P3 component amplitude had maximal values in the temporal leads. The temporal visual system seems to play a domineering part in the tasks of spatial and non-spatial attention to stimuli in humans. PMID- 14758659 TI - [Porcine kidney extract contains a specific inhibitor of the ouabain-sensitive alpha2-isoform of Na, K-ATPase present in rat diaphragm fibres]. AB - In experiments on isolated rat diaphragm muscle, acetylcholine (100 nmol/l) hyperpolarized muscle fibres due to activation of the alpha 2 isoform of Na,K ATPase. This hyperpolarization was blocked in a dose-dependent manner by ouabain (K0.5 = 8 +/- 4 nmol/l) as well as by a solution of porcine kidney extract (10 kDa cut-off filtration), with the K0.5 approximately equal to a 1:20,000-fold dilution. The inhibitory activity of the developed slowly over a period of 3 hours and, in contrast to ouabain, was still present after 1 hour of washing. Ouabain, but not the extract, inhibits Rb+ uptake in human erythrocytes that only express the alpha = 1 isoform of Na, K-ATPase. Our data suggest that in rat skeletal muscle the alpha 1 isoform of Na,K-ATPase is primarily responsible for ionic homeostasis, while the alpha 2 isoform provides a "regulatable" function and may be controlled by cholinergic stimulation and/or endogenous digitalis-like factors (EDLFs). Porcine kidney extract contains a factor (M. W. < 10 kDa) that selectively inhibits the rat alpha 2 isoform and differs from ouabain. Our experimental protocol can be used as a highly sensitive physiological assay for factors that selectively inhibit the alpha 2 isoform of Na,K-ATPase. PMID- 14758660 TI - [Pineal peptides in the regulation of milk-ejection reflex in lactating rats]. AB - The effects of pineal peptides (mol. mass 1-4 kDa) intranasal infusions on some parameters of milk-ejection reflex were investigated. Peptides were extracted from dairy-cattle pineal glands. Pineal peptides increase body weight, levels of water intake, plasma prolactin concentration and milk yield in rats when infusing daily in dose 1 microgram/kg from the third day of lactation. On 9th and 12th days of lactation during 1-hour nursing seance the significantly greater number of reflective milk ejections were found. When 1 hour before the peptides infusion rats were intraperitoneally injected by rabbit antiserum to oxytocin (200 microliters at dilution of 1:20) the effects of pineal peptides were significantly less expressive or were absent at all. Using enzymimmunoassay it was demonstrated that there were greater increasing of oxytocin content in pineal gland in suckling-induced oxytocin release from neurohypophysial system in the chronic pineal peptides-treated female rats compared with control. This effect was absent when rats were injected by oxytocin antibodies. These data suggest that pineal peptides can participate in forming of reflect oxytocin release pattern. This pattern is initiated by suckling and is limited by oxytocin content in blood. PMID- 14758661 TI - [Effect of pineal peptides on the adrenal gland cortex glucocorticoid function and behavior in rats orally immunized with ovalbumin]. AB - The influence of intraperitoneal injections of pineal peptides (5 mg/100 g of body mass during first five days of three weeks' oral immunization by ovalbumin) on the rats' behavior in the "open field" tests and on the blood corticosterone level, was investigated. It was found out that rats' oral immunization resulted in increasing of secretion activity of Peyer's patches antibody-producing cells, in decreasing of blood leukocyte cytokine-producing activity, in depression of the searching behavior and locomotor activity and in a significant (p < 0.05) lowering of the blood corticosterone level after 15 minutes in the "open field" tests. The pineal peptide injections caused an intensification of humoral immune response, a more obvious suppression of locomotor activity and searching behavior, and a significant decrease of the corticosterone level compared to the animals intraperitoneally injected by physiological solution. These data indicate that immuno-stimulative effect of pineal peptides combines with their ability to decrease glucocorticoid hormone secretion during stress-reaction. PMID- 14758662 TI - [Mechanisms of human adaptation to hypoxia]. AB - The effects of adaptation to cold-and-hypoxic exposure on the cardiovascular system, lipid peroxidation and concentrations of adaptogenesis involved hormones were studied in male students. The two weeks cold- and hypoxic training was shown to be accompanied by a significant increase of apnea duration, reduced velocity of bradycardia development and a more rapid ECG post-cold and- hypoxic exposure normalization, as well as by inhibition of activation of adrenal cortex and thyroid gland after stress of different nature. The changes of the character of influences between the indices under study, were demonstrated. The correlation analysis showed an increase of the human's adaptive potential and a decrease of its dependence on the adrenal cortex hormones. PMID- 14758663 TI - [Steroid hormones in the regulation of migration in fishes (study of the Russian sturgeon)]. AB - Serum steroids profiles were determined at different stages of the migratory cycle of Russian sturgeon. In the sea period, before gonads maturation cortisol and sex steroids levels were comparatively low. Elevation of cortisol and testosterone concentrations occurred at the stage of preparation to migration. Significant increase of cortisol and testosterone levels happened at the beginning of the river period of anadromous migration in spring both in the spring forms near to maturity in this period and in the winter forms which were far from maturity. By exception of the river period of anadromous migration of the winter form and sexual cycle completion in the fish farm in the mouth of river (10-11 months), serum cortisol and testosterone concentrations dropped sharply. The data obtained as well as the data on other diadromous fishes indicate a possible role of steroids, especially cortisol and testosterone, in metabolism and migration regulation in sturgeons. PMID- 14758664 TI - [Serum steroid hormones in female Russian sturgeon (Acipenser gueldenstaedtii Br.) with normal oocyte maturation and with gonadal function disorders after hormonal stimulation]. AB - Blood serum cortisol, testosterone, and 11-ketotestosterone levels were determined by ELISA methods in female Russian sturgeon after hormonal stimulation for induction of ovulation. All females were divided into 5 groups depending on their initial reproductive status and response to hormonal stimulation; 1) normal ovulation, 2) ovulation followed by release of non-viable eggs, 3) normal state of ovary without ovulation, 4) lack of ovulation, early stage of oocyte resorption, 5) lack of ovulation, advanced stage of oocyte resorption. Cortisol levels in all five groups did not differed significantly. Testosterone levels were low (15-24 ng/ml) in females after ovulation and in females without response to hormonal stimulation because of early stage of oocyte resorption. Non-matured females with normal state of oocytes had significantly higher testosterone levels -82 +/- 16 ng/ml. Non-matured females with advanced stage of resorption were subdivided into 2 subgroups--with low (approximately 7 ng/ml) and high (> 100 ng/ml) levels of serum testosterone. 11-ketotestosterone levels were similar in all investigated groups of sturgeon. PMID- 14758665 TI - [Changes in temporal parameters of compression stimuli in milk ejection during feeding a baby]. AB - The temporal changes of compression stimuli exerted by baby on the areola-nipple complex during the entire suckling period were investigated in 6 mother-baby pairs of the 4-6-day lactation. It was found that compression stimuli had a triangle form. The duration of the stimuli and inter-stimuli interval varied from 0.3 s to 0.8 s and from 0.1 s to 0.25 s, respectively. The compression stimuli were organized in bursts and a pause of variable duration between the bursts. The mean frequency of the compression stimuli in the bursts was within the range 0.9 imp/s-1.7 imp/s. The average frequency of the entire suckling period in babies was within the range 0.9 +/- 0.3 imp/s-1.1 +/- 0.1 imp/s. Implications of these findings in the suckling neonates are discussed. PMID- 14758666 TI - [The role of calcium and potassium ions in the response of the mouse mammary secretory and myoepithelial cells to treatment with oxytocin]. AB - Experiments were carried out in lactating white mice. Removal of Ca(2+)-ions from the perfusion solution reduced the amplitude and duration of the membrane potential changes in the secretory cells in the mammary gland alveolus. The extra and intracellular Ca(2+)-ions participate in development of contraction responses of myoepithelial cells. Removal of K(+)-ions from perfusion solution and increase of K(+)-ions concentration in the medium to 20 mmol/l inhibit the development of response of secretory cells to oxytocin action. These changes in K(+)-ions concentration do not affect the contractile response of the myoepithelial cells. The findings suggest that there are essential differences in the participation of Ca(2+)- and K(+)-ions in mechanisms of the mammary secretory and myoepithelial cells responses to oxytocin action. PMID- 14758667 TI - [Spectral characteristics of vowel-like sounds in children of the first year of life and speech development]. AB - To study the continuity in speech development, an investigation of vowel-like sounds recorded in the course of longitudinal research of speech formation was carried out in 12 infants beginning with the first month up to 12 months of their life. It was revealed that features ensuring separation of vowel-like sounds are specific by their amplitude relation and frequency location of the most expressed spectral maxima, including the maxima corresponding to the fundamental frequency. As fundamental frequency increased, the vowel-like sounds [a], [u], [i] began to exhibit specific changes of the amplitude ratios of the spectral maxima. It was established that peculiarities of frequency positions of spectral maxima and relations of their amplitudes may be the very features on the bases of which children compare their own vowel-like sounds to the vowels of the adults in sound imitation. These findings and literature data corroborated the opinion of continuity in speech development, beginning with the early preverbal vocalizations. PMID- 14758668 TI - [Lipid peroxidation in rabbit and rat visual system structures]. AB - In our work, the lipid peroxidation (LPO) in the retina, optic chiasma, and visual cortex of rat and rabbit brain was investigated. The contents of the LPO products (diene conjugates, triene conjugates, TBA-reactive products, Schiff bases) and oxidation index (calculated as 232/2 15) were similar in the retina and visual brain cortex of rats. In vivo, lipid oxidation in the optic chiasma was higher as compared with two other parts of visual tract. The similar data were obtained in our experiments with rabbit's visual tract. The sensitivity of tissues to peroxidation in vitro was studied in homogenates incubated with 0.2 mM ascorbate and 10 mkM FeSO4 for 20 min at 37 degrees C. The results of these experiments deviated from the data obtained in vivo, namely: the LPO in optic chiasma was lower than in the retina and the brain cortex. This data are in compliance with lipid composition of investigated parts of the visual tract of both animals. In our opinion, the high level of LPO in optic chiasma demonstrated in vivo is due to low antioxidants level in this part of the visual tract. Our findings also indicate that LPO in retina both in vivo and in vitro experiments are similar to those in the brain cortex and may be attributed to similar lipid composition and activity of antioxidant enzymes (such as superoxiddismutasa and glutathionereductase). PMID- 14758669 TI - [Absence of the effect of opioid peptides on muscarine receptors in the frog vestibular apparatus]. AB - We studied the effects of opioid peptide leu-enkephaline, a specific antagonist of acetylcholine receptors atropine, and non-selective opiate antagonist naloxone on synaptic transmission and responses evoked by acetylcholine in semicircular organs of the frog. A decrease in frequency of acetylcholine (0.1-5.0 microM) responses under leu-enkephaline (10 nM) id not differ from the frequency decline induced by leu-enkephaline alone. Atropine (1 microM) left the response to leu enkephaline intact while blocking the excitatory effect of acetylcholine. No modification of the acetylcholine response under leu-enkephaline was observed in the presence of naloxone (1 microM). The findings suggest that no interaction exists between the acetylcholine-mediated excitatory action on resting activity in the isolated semicircular canal preparation and the suppressive action of leu enkephaline. PMID- 14758670 TI - [The effect of neurochemical damage of dopaminergic terminals in early ontogenesis on the behavior of adult rats]. AB - 6-Hydroxydopamine (75 mkg), producing selective degeneration of dopaminergic neurons in the brain, was injected intraamniotically to every rat fetus on 13th or 17th day of mother pregnancy. The other experiment was performed, when 6 hydroxydopamine administered i.p. to newborn rats on 4th or 10th day of life. All rats were growing, and several dopamine-dependent behaviors were investigated in adult animals: open field, rotation behavior, anxiety in elevated plus maze, conditioned placed preference, differentiation of novel and known alleys of Y maze, aggressive behavior in intruder-resident test, selfs-stimulation of lateral hypothalamus. Prenatal administration of 6-hydroxydopamine initiated rotations and stereotypy, decreased anxiety in elevated maze, reduced reinforcing properties of amphetamine in place preference test, disturbed differentiation of novel and known alley in Y-maze, high aggression and decreased self-stimulation in less degree that postnatal injection of neurotoxin. Therefore, the early postnatal period is more sensitive to neurotoxin action than prenatal period of development. This phenomenon is connected with critical periods of development of dopaminergic system in ontogeny. PMID- 14758672 TI - A primer on oncology for the primary care PA. PMID- 14758671 TI - [140 years since foundation of the department of general physiology in the St. Petersburg University by Filipp Vasil'evich Ovsiannikov ]. PMID- 14758673 TI - Cancer screening in primary care. PMID- 14758674 TI - Novel cancer therapies: targeting the molecules. AB - Translational research is providing the means to develop a growing arsenal of targeted oncologic therapies (see Table 2, page 31). In practice, some of these drugs have completely changed the treatment pathway for common cancers. Many patients can be treated with greater efficacy and fewer side effects than traditional therapies allowed. Some patients with poor performance status have been able to undergo treatment when they would otherwise not have qualified for it. Some agents have not been shown to improve overall survival, but they do offer the possibility for improvement in quality of life. Although not miracle cures, these current targeted therapies offer proof of principle for scientists and hope for millions of patients. PMID- 14758675 TI - Reacting swiftly to oncologic emergencies. PMID- 14758676 TI - Symptom control in advanced cancer: meeting the challenge. PMID- 14758677 TI - PA residency accreditation: why we need it now. PMID- 14758678 TI - Current therapies for pulmonary artery hypertension. PMID- 14758679 TI - Acute viral hemorrhagic cystitis. Alarming symptoms with a benign cause. PMID- 14758680 TI - The AAAAI adherence guidelines: taking control of asthma therapy. PMID- 14758681 TI - Diagnosing and managing first carpometacarpal joint arthritis. PMID- 14758682 TI - Varicella in an adult. PMID- 14758683 TI - The quiet killer that took my friend. PMID- 14758684 TI - Endometriosis. A common cause of infertility and pelvic pain. PMID- 14758685 TI - Postmastectomy breast reconstruction. Helping patients decide whether--and when. PMID- 14758686 TI - Diagnostic and management strategies for pregnant women with back pain. PMID- 14758687 TI - Mastering mammography. When to order, and what to make of the results. PMID- 14758688 TI - Understanding the role of sentinel lymph node biopsy in breast cancer and melanoma. PMID- 14758689 TI - Draining a cyst or abscess in a Bartholin's gland with a Word catheter. PMID- 14758690 TI - Are we listening to our patients in pain? PMID- 14758691 TI - Scoliosis: a straightforward approach to the curved spine. PMID- 14758692 TI - When to screen for factor V Leiden. PMID- 14758693 TI - Smoking cessation. A review of current treatments. PMID- 14758694 TI - Fracture of the tibial plateau. PMID- 14758695 TI - Ecology of a stream from upper Parana River basin inhabited by Aspidoras fuscoguttatus Nijssen & Isbrucker, 1976 (Siluriformes, Callichthyidae). AB - Most information on catfishes of the genus Aspidoras has focused species kept in aquaria, in non-natural conditions. Biotic and abiotic parameters of environment inhabited by Aspidoras fuscoguttatus, associated ichthyofauna, as well as strategies involved in the successful occupation of different microhabitats in a first order stream, located in Sao Jose do Rio Preto, Sao Paulo State, were investigated. Data collection was monthly performed at three segments (headwaters, middle course, and mouth) from August 1999 to July 2000. The greatest values of water temperature, conductivity, turbidity, alkalinity, width, depth, and current velocity were obtained in the rainy season. Dissolved oxygen levels showed significant differences among three segments in all sampling months, except for September. Substrate composition was predominantly sandy, followed by silt and clay in middle course and mouth in rainy as much as in dry seasons. In headwaters a large increase of the sandy fraction occurred in the rainy season. In the marginal and aquatic vegetation 30 species of macrophytes and 5 of macroalgae were identified. The fauna included tadpoles of Anura and macroinvertebrates (Mollusca, Crustacea, Arachnida, and 8 orders of Insecta). A. fuscoguttatus was found throughout the entire stream in sympatry with 6 fish species. In middle course, cluster analysis revealed an association between A. fuscoguttatus immatures and low depth, suggesting a probable function of depth in that stage of ontogenetic development of catfishes. The results indicate that A. fuscoguttatus present plasticity in terms of microhabitat, including air breathing behavior in hypoxic conditions, previously unknown in genus Aspidoras. PMID- 14758696 TI - Methodological tests of a heterotrophy index for aquatic ecosystems. AB - Experiments in glucose mineralization were carried out to investigate the effects caused by natural forcing functions on both the decomposition rates and heterotrophy capacity of aquatic ecosystems. In addition, the methodology used could show connections between mineralization rates measured in both laboratory and field work with those measured in aquatic ecosystems. Water samples from Infernao lagoon (21 degrees 35'S and 47 degrees 51'W) were collected, filtered, enriched with glucose, and incubated under aerobic and anaerobic conditions. The glucose concentration variation, dissolved oxygen (DO) consumption, pH, electric conductivity, and total CO2 amount in the water were determined for sixteen days. In the period with intense oxygen consumption there was also an evident glucose demand and the dissolved oxygen consumption rate was approximately the same as that for glucose mineralization. The process in the aerobic chambers was 2.2 times faster than that in the anaerobic chambers. An initial acidification of the water samples, probably due to microbial carbonic acid liberation, was noted. A rise in pH values was also observed at the end of the process. The electric conductivity was low for both aerobic and anaerobic chambers, indicating a probable ion uptake by microbial organisms due to the presence of carbon sources. The glucose content variations corresponded to both CO2 formation and dissolved oxygen consumption. It was estimated that 19.4% of the initial glucose content turned into CO2 and the remaining 80.6% into humic compounds and microbial biomass. This experiment showed that glucose can be used as a substrate indicating the heterotrophy of a given aquatic ecosystem. PMID- 14758697 TI - Spatial and temporal distribution of "curvina" larvae (Plagioscion squamosissimus Heckel, 1840) and its relationship to some environmental variables in the Upper Parana River floodplain, Brazil. AB - With the objective of studying the spatial and temporal distribution of "curvina" Plagioscion squamosissimus larvae and verifying respective spawning sites, monthly sampling was carried out from March 1993 to February 1994 on the Upper Parana River floodplain. Using a conical-cylindrical plankton net (0.5 mm mesh), samples were taken from the surface of the water bodies, and in the marginal vegetation a strainer and a dragnet ("picare") were used. Water samples were collected for environmental variable analysis (temperature, dissolved oxygen, water pH, and electrical conductivity). Water level data were obtained from hydrometric stations. High average density of larvae was found from December 1993 to February 1994 due to high temperature and water level. The Baia sub-area was shown to be the most important in the larvae catch, probably due to its semi lotic characteristics. Larvae size distribution was shown to be more frequent in the 4.0 mm standard length class, indicating that the sampling stations were next to the spawning sites. PMID- 14758698 TI - Patterns of infestation by chigger mites in four diurnal lizard species from a Restinga habitat (Jurubatiba) of southeastern Brazil. AB - We studied the parasitism by larvae of the chigger mite Eutrombicula alfreddugesi on the lizard community of Restinga de Jurubatiba, Rio de Janeiro State, Southeastern Brazil. We investigated the patterns of infestation (prevalence and intensity) of chigger mites in four sympatric lizards: Tropidurus torquatus, Mabuya agilis, M. macrorhyncha and Cnemidophorus littoralis. All lizards collected were checked for the presence of mites, which were counted under stereomicroscope. We tested the relationship between intensity of infestation and lizard body size for each species using regression analysis. The prevalences and mean intensities (+ one standard deviation) of infestation on each host species were, respectively: 100%; 86.4 + 94.6 in T. torquatus (n = 62); 100%; 20.9 + 9.3 in M. agilis (n = 7); 100%; 11.1 + 13.1 in M. macrorhyncha (n = 12); and 95.2%; 19.1 + 16.8 in C. littoralis (n = 21). Only for C. littoralis did body size significantly affect the intensity of infestation (r = 0.27, p = 0.02). For all lizard species, the body parts where chiggers occurred with the highest intensity were those of skin folds and joint regions. PMID- 14758699 TI - Distribution of sandflies (Diptera: Phlebotominae) in forest remnants and adjacent matrix habitats in Brazilian Amazonia. AB - We studied the distribution of sandflies (Diptera: Phlebotominae)--insect vectors of several diseases, including leishmaniasis--at the interface between primary forest and cattle pasture and between primary forest and secondary forest (< 15 yr old) in Southern Brazilian Amazonia. Sandflies were collected by using a combination of light traps and traps having vertebrates as baits. Strong differences in abundance and species richness were found between primary forests and pastures. Very few sandfly species were found in the pastures, and those that were found generally occurred at lower densities when compared to the adjacent forest. At least one species (Lutzomyia lainsoni), however, can become extremely abundant in pastures, possibly depending on the presence of cattle and water bodies. Differences between primary forests and secondary forests were not so strong, although the latter usually had fewer species and lower population abundances. No species were exclusively found in pastures or secondary forests; the species present in these two habitats were a subset of those found in primary forests. The distance to the edge did not affect the abundance, richness and composition of sandfly species in primary forests. The abundance and richness of sandflies, however, was greater in forest edges facing pastures than those facing secondary forests. This pattern could not be explained by an influx of species and individuals from the adjacent pasture, suggesting the existence of in situ differences between the different types of forest edges studied. PMID- 14758700 TI - New observations on frustule morphology of Eupodiscus radiatus Bailey and Fryxelliella floridana Prasad. AB - A study of the diatoms Eupodiscus radiatus Bailey and Fryxelliella floridana Prasad, mainly focussing on the mantle and cingulum, provided new morphological information. In E. radiatus dendritic structures and two types of a palisade-like structure fixed to silica rings were found on the lower valve mantle. Cingulum presented 1-3 bands with areolae arranged in decussate rows. Furthermore, the pars interior of the valvocopula is fimbriate; and the external openings of the rimoportulae are located along the rim of the scalloped extension. The valvocopula of F. floridana is open and its copula is ligulate. Both bands possess poroid areolae similar in size to the cribral pores on the valve face. The genus Eupodiscus is compared to Fryxelliella, based on material sampled in estuaries of Southern Brazil. PMID- 14758701 TI - Spatial patterns and relations with site factors in a campos grassland under grazing. AB - Spatial distribution patterns and their relations with environmental factors at different scales were identified in ca. 100-ha grassland under cattle and sheep grazing, in Eldorado do Sul, RS, Brazil (30 degrees 05'S, 51 degrees 40'W). The field survey used 138 0.5 x 0.5 m quadrats located systematically on transects along relief gradients. The quadrats were arranged in groups of 3 contiguous quadrats, which were pooled for the analysis, thus forming 46 quadrats 1.5 x 0.5 m, in this way defining two observation scales. Vegetation description involved recording the presence and the visual estimation of cover-abundance of species in each quadrat. A total of 148 species belonging to 30 families was detected. The environmental conditions at each site were described by 30 variables related to soil chemical and physical properties, slope, exposure and relief position. Data analysis used cluster analysis, evaluation of group partition sharpness, ordination, significance of ordination axes, evaluation of environmental congruence and randomization testing. The results of the analysis with 46 quadrats supported those found with 138 quadrats. The vegetation patterns in the study area are associated to relief position and other related factors such as soil moisture. Two clearly defined grassland community types were detected, one occurring on the slopes and another on the wet lowlands. PMID- 14758702 TI - Evolutionary implications of intra- and interspecific molecular variability of pathogenesis-related proteins. AB - We have examined phylogenetic relationships in seven pathogenesis-related (PR) protein families. Within-family comparisons involved 79 species, 166 amino acid sequences, and 1,791 sites. For 37 species, 124 different PR isoforms were identified (an average of 3.3 per species). Thirty-one of the 37 species investigated tended to cluster together (84%). Of the 17 clusters distinguished in the seven phylogenetic trees, 10 (59%) were in agreement with their taxonomic status, ascertained at the family level. The strong similarities among the intraspecific forms, as compared to interspecific differences, argue for some kind of gene conversion, but the rare occurrence of widely different isoforms also suggests diversifying selection. PRs 1, 6, and 4 seem to be less differentiated than PRs 3, 2, 10, and 5. PMID- 14758703 TI - Experimentally induced heat- and cold-shock tolerance in adult Panstrongylus megistus (Burmeister) (Hemiptera, Reduviidae). AB - The survival rate of domestic male and female adult Panstrongylus megistus was studied after sequential heat and cold shocks in order to investigate shock tolerance compared to that previously reported for nymphs. Sequential shocks were such that a milder shock (0 degree C, 5 degrees C, 35 degrees C, or 40 degrees C for 1 h) preceded a severe one (0 degree C or 40 degrees C for 12 h), separated by intervals of 8, 18, 24, and 72 h at 28 degrees C (control temperature). The preliminary thermal shock induced tolerance to the more severe one, although tolerance intensity depended on the initial shock temperature and the interval between treatments. Despite the observed tolerance, the survival rate for insects subjected to both shocks decreased when compared to that of individuals subjected to a single mild shock. When tolerance differed with sex, females showed greater values than males. In contrast to the response detected in nymphs, for which higher heat tolerance values were sustained for intervals of up to 24 h (preliminary shock, 35 degrees C) or even longer (preliminary shock, 40 degrees C) between sequential shocks, significant values were verified in adults only for shock intervals of up to 8 h (preliminary shock, 40 degrees C). While findings for nymphs exhibited considerable cold-shock tolerance under conditions in which preliminary shocks were given at 5 degrees C or 0 degree C and the periods between shocks were up to 72 h long, the adults were shown to be capable of acquiring a substantial tolerance response to a more severe cold shock only when the preliminary shock was given at 0 degree C and shock interval surpassed 18 h. It is assumed that the mechanisms involved in the cellular protection of P. megistus under sequential temperature shocks (heat shock protein action?) may loose effectiveness with insect development. PMID- 14758704 TI - Thermal evidence of the invasion of a stingless bee nest by a mammal. AB - Melipona bicolor, an inhabitant of the Atlantic Rainforest, nidifies in hollows of live or dead trees. In order to study thermoregulation of a nest of this species, a temperature data logger was installed inside a hollow tree. After this, an intruder dug a hole, invaded the nest, and probably consumed its honey, pollen and bees, having remained there during three days. Thermal evidence and its behavior allowed the delimitation of a small number of suspects, which we analyzed here. The intruder was a small mammal, predominantly nocturnal, that takes shelter in burrows, probably the yellow armadillo (Euphractus sexcinctus). Other evidence, if collected immediately after invasion, could precisely indicate precisely the species. PMID- 14758705 TI - Biology of Brontocoris tabidus (Heteroptera: Pentatomidae) fed with Musca domestica (Diptera: Muscidae) larvae. AB - Development and reproductive performance of Brontocoris tabidus (Signoret) (Heteroptera: Pentatomidae) fed with Musca domestica L. (Diptera: Muscidae) larvae was evaluated. Duration of the nymphal phase of this predator was 31.2 days, with a 44.1% survival rate. Preoviposition and egg incubation periods and number of eggs/egg mass were 8.14 days, 5.6 days, and 45.4, respectively. B. tabidus presented 44.7% egg viability. Female longevity of this predator was 18.4 days. Results are discussed based on this prey use in mass rearing and maintenance of colonies of B. tabidus under laboratory conditions. PMID- 14758706 TI - Population biology and distribution of the tanaid Kalliapseudes schubarti Mane Garzon, 1949, in an intertidal flat in southeastern Brazil. AB - The population biology and the spatial and temporal distribution of Kalliapseudes schubarti Mane-Garzon, 1949, a common tanaidacean in mud flats and estuaries in southern and southeastern Brazil, was studied in the Araca region, Sao Sebastiao (SP), Brazil. This species showed a clustered dispersion in the area and the individuals were concentrated in the superficial sediment layer (5 cm). Higher densities of K. schubarti were recorded in areas characterized by moderately sorted fine sediment. Multiple regression analysis revealed a positive influence of the organic matter contents and a negative effect of the silt-clay contents on the abundance of K. schubarti. This species showed a marked temporal variation with very low abundance in winter and fall (March to August). Sexual dimorphism was evidenced with males being larger than females. Ovigerous females were also larger than pre-ovigerous ones. Sex ratio was skewed towards females. Seven cohorts were identified during the sampling period, the estimated longevity was 12 months, and no seasonal oscillation in growth was evidenced. The continuous reproduction, as evidenced by the presence of larval phases (manca II and neutron) and reproductive females throughout the year, and high fecundity among the tanaids associated with fast growth and limited longevity support the case for the opportunistic life strategy suggested for this species in the literature. PMID- 14758707 TI - Studies on mineral nutrition of the coffee plant (Coffea arabica L. cv. Catuai Vermelho). LXIV. Remobilization and re-utilization of nitrogen and potassium by normal and deficient plants. AB - Remobilization and re-utilization of 15N and 85Rb labelled nitrogen and potassium reserves for new growth and fruit formation was studied under greenhouse conditions using both normal and deficient young coffee plants. It was found that K reserves are used in higher proportion than is stored N by fruits and other organs. The export of N by organs of residence in the normal plants obeyed the following proportions of the total: leaves 47%-58%, branches and flower buds 21% 27%, roots 21%-32%. The corresponding figures in the case of deficient plants were: leaves 49%-65%, branches and flower buds 21%-27%, roots 14%-25%. Re utilization of K took place in the following proportions in the normal plants: leaves 54%-64%, branches and flower buds 20%-21%, roots 30%-41%. In K deficient plants the figures were: leaves 62%-79%, branches and flower buds 1.2%-4.4%, roots 20%-33%. In tissues formed after the initiation of flowering buds, the demand for N is met by reserves as follows: normal plants: fruits 20.6%-24.8%, leaves 15.6%-19.4%, twigs 19%-20.5%; deficient plants: fruits 43.5%-48.5%, leaves 48.1%-51.9%, twigs 46%-53%. The K needs for new tissues are met in the order: normal plants: fruits 40%-45.8%, leaves 27%-37.6%, twigs 26%-33.1%; deficient plants: fruits 65.7%-81.5%, leaves 52.6%-68.4%, twigs 62%-86.1%. Fruits represent the main sink for both N and K. Reutilization of both elements is higher in the case of deficient plants. PMID- 14758708 TI - Bacterial phagocytosis by macrophage of autogenous splenic implant. AB - Autogenous splenic implant seems to be the only alternative for preservation of splenic tissue after total splenectomy. This work was carried out to analyze the morphologic regeneration of autotransplanted splenic tissue in Wistar rats and to determine the bacterial phagocytic function of their macrophages. We utilized an experimental model with thirty-two rats, of both sexes, submitted to total splenectomy combined with autotransplantation in greater omentum of slices of the whole spleen mass. The animals were divided into two groups: I--young rats weighing 100 to 150 g; and II--adult rats weighing 250 to 300 g. Sixteen weeks later animals were intravenously inoculated with a suspension of Escherichia coli AB1157. Twenty minutes after inoculation, the animals were sacrificed and the splenic autotransplants were removed for morphological study. There was regeneration of autotransplanted splenic tissue in all animals. A similar morphological aspect among all animals was observed, with splenic tissue showing red and white pulps, lymphoid follicles, and marginal zone, with a moderate architectural disarrangement. Macrophages containing gram-negative bacterial aggregates as well as macrophages with hemosiderin pigments within the cytoplasm were observed. Blood vessels showed preserved walls, with no signs of vasculitis or thrombosis. The present results suggest that autogenous splenic implants in the greater omentum of the rat acquire the macro- and microscopic architecture of a normal spleen, with reduced dimensions, and preserve bacterial phagocyte function. PMID- 14758709 TI - A new nematode species Goezia leporini n. sp. (Anisakidae) from cultured freshwater fish Leporinus macrocephalus (Anostomidae) in Brazil. AB - This paper describes nematode infection in the cultured freshwater fish Leporinus macrocephalus (Osteichthyses: Anostomidae) collected at Batatais, Sao Paulo State, Brazil. Of a total of 32 examined fish, 21 (65%) were infected with Goezia leporini n. sp. (Nematoda: Anisakidae) with mean infection intensity of 4.1 parasites. The nematodes presented total length greater than G intermedia, G. holmesi, G. pelagia, G. minuta, G. kliksi, G. sinamora, G. nonipapillata, G. alii, G. moraveci, G. brasiliensis, and G. brevicaeca. The main difference was a great number of preanal papillae in males when compared to G. brasiliensis and G. brevicaeca. The present description also differs from that of G. brasiliensis with respect to spicule length and distance of vulva from the anterior extremity. PMID- 14758710 TI - Bioacoustic analysis of advertisement call in Hyla nana and Hyla sanborni (Anura, Hylidae) in Botucatu, Sao Paulo, Brazil. AB - Vocal communication traits of Hyla nana and Hyla sanborni, of the nana species group, were studied from August 1997 until June 1999 in two large permanent ponds located in an open field in Botucatu, Sao Paulo State. One hundred thirty-one individuals, 71 of H. nana and 58 of H. sanborni, were recorded in the beginning of their vocalization activity and during chorus vocalization. The rhythms of sound emission on the two occasions were different. An advertisement call consists in a consecutive series of simple notes in rapid succession. Both species have two types of notes in their advertisement calls, here named types A and B. Type A notes are introductory and have a longer and higher pulse number and are emitted more frequently in the beginning of vocalization activity. Introductory notes are the first to be emitted in chorus activity. Type B notes are secondary, of shorter duration and lower pulse number, and are emitted during chorus vocalization. The notes of both types differ significantly in their temporal structure. Both species present acoustic segregation in both spectral and temporal structure. PMID- 14758711 TI - Differential leukocyte counts in "dourado", Salminus maxillosus Valenciennes, 1840, from the Mogi-Guacu River, Pirassununga, SP. AB - From August, 1996 to December, 1997, 293 of "dourado" specimens, Salminus maxillosus (Valencienes, 1840), of various sizes, were caught in Mogi-Guacu River, Emas Falls, for hematologic studies. Total weight (Wt in g) and length (Lt in cm) were taken for each individual animal. Smears were prepared from blood samples and utilized for differential leukocyte counts (lymphocytes, neutrophils, monocytes, eosinophils, special granulocytic cell and immature cells). The mean percentages of leukocytes were determined according to sex and stage of gonadal maturation (immature, in maturation, mature, spent and resting). Significant differences between male and female occurred only for the special granulocytic cell. In analyzing the leukocyte profile during gonadal development, only female showed significant differences in mean percentages of lymphocytes, neutrophils, monocytes and eosinophils. PMID- 14758712 TI - Current state and projection of the probable original vegetation of the Sao Carlos region of Sao Paulo State, Brazil. AB - A map of the native vegetation remaining in Sao Carlos County was built based on aerial images, satellite images, and field observations, and a projection of the probable original vegetation was made by checking it against soil and relief surveys. The existing vegetation is very fragmented and improverished, consisting predominantly of cerrados (savanna vegetation of various physiognomies), semideciduous and riparian forest, and regeneration areas. Araucaria angustifolia (Bertol.) Kuntze, found in patches inside the semideciduous forest beginning at a minimum altitude of 850 m, has practically disappeared. By evaluating areas on the map for different forms of vegetation, we obtained the following results for original coverage: 27% cerrado (sparsely arboreal and short-shrub savanna, and wet meadows); 16% cerradao (arboreal savanna); 55% semideciduous and riparian forests; and 2% forest with A. angustifolia. There are now 2% cerrados; 2.5% cerradao; 1% semideciduous forest and riparian forests; 1.5% regeneration areas; and 0% forest with A. angustifolia. PMID- 14758713 TI - Daily activity of four tropical intertidal hermit crabs from southeastern Brazil. AB - This study describes the daily activity in a simulated high tide situation of four species of hermit crabs (Pagurus criniticornis, Clibanarius antillensis, C. sclopetarius, and C. vittatus) that coexist in an intertidal flat in southeastern Brazil. Observations were done in two-hour intervals during two subsequent days (48 h) in three replicate pools with thirty crabs each. Among species (between and within genera) there was an evident variation in activity patterns, of which three could be distinguished. The circadian activity patterns of C. antillensis and C. vittatus could be characterized as evening and nocturnal, with resting peaks during the morning and afternoon. The circadian activity pattern of C. sclopetarius was characterized by two marked peaks of inactivity, corresponding to dawn and evening, which could represent an intrinsic association with the semi lunar tidal cycles of the study area. Pagurus criniticornis showed high activity not influenced by day/night conditions during the entire observed period. These activity pattern variations of the studied hermit crabs should be taken into account in designing further experiments. More precise and accurate interspecific behavioral comparisons among species could be achieved in nocturnal experiments, the high activity period of all species. PMID- 14758714 TI - A method for quantifying bird colonies in sand bars via GPS. PMID- 14758715 TI - "Out-of-field" effects of head-localized proton irradiation on peripheral immune parameters. AB - The heads of Sprague-Dawley rats were irradiated with protons to total doses of 1.5, 3 and 4 Gy and euthanized 9-10 days later. Significant dose-dependent decreases were noted in thymus mass. Lymphocyte and platelet numbers were significantly reduced in blood. Flow cytometric analysis of blood and spleen showed that CD3+ T, CD3+/CD4+ TH, and CD3+/CD8+ TC cell numbers were low and proportions were significantly altered by radiation. CD4:CD8 ratios and CD45R+ B lymphocytes were unaffected. Spontaneous blastogenesis of blood and spleen leukocytes was significantly increased by radiation. Plasma TGF-beta 1 level in irradiated rats was consistently, but not significantly, higher than in non irradiated animals. T and B cell proportions in lymph nodes from irradiated animals were similar to non-irradiated controls. Bone marrow from all irradiated groups had high CD90+/Gran+ cell numbers. The data show that head-localized proton irradiation at relatively modest doses can profoundly influence systemic distribution and composition of lymphocyte populations. The data also suggest that immune modulation induced by localized proton, as well as other forms of radiation, should be taken into consideration when evaluating adjunctive immunotherapies in patients receiving radiotherapy. PMID- 14758716 TI - The effect of co-administration of simvastatin and alcohol in rats. AB - INTRODUCTION: The effect of chronic co-administration of alcohol (Alc) and lipid lowering drugs on hepatic function has not been extensively evaluated. We studied the effects of administering Alc together with a 3-hydroxy-3 methylglutaryl coenzyme A reductase inhibitor [simvastatin (S)]. MATERIALS AND METHODS: Male Wistar rats (8 weeks old) were randomized and divided into 4 groups of 10 each. They were fed (once a day) via a stomach tube with: 1) 2 ml of olive oil; group Oil, 2) with Oil + 2 ml of 25% v/v pure Alc in water; group Alc + Oil, 3) with Oil + S (65 micrograms/100 g body weight); group S + Oil, 4) with Oil + Alc + S; group S + Alc + Oil. Another 13 male Wistar rats were only fed a standard laboratory diet (control group). After 8 weeks blood samples were drawn and the livers were removed. Blood glucose, alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (AP), total protein, albumin, total cholesterol (TC) and triglycerides (TG) were measured. Liver histopathology was also assessed. RESULTS: Differences were found between the control group and tube fed groups in glucose (p < 0.001). No differences were found among tube-fed groups in blood glucose, ALT, total protein, albumin, AP and TC. AST activity was significantly higher in the Alc + Oil than in the Oil or S + Oil groups (p < 0.001 for both comparisons) demonstrating the effect of Alc on AST. The AST did not differ significantly in the Oil or S + Oil groups indicating a lack of effect of S. Furthermore, S significantly reduced the Alc-induced increase in AST (Alc + Oil vs S + Alc + Oil; p = 0.042). The TG concentration was significantly higher in the Alc + Oil group compared to the Oil, S + Oil and S + Alc + Oil groups (p = 0.02). Therefore, S significantly decreased the alcoholinduced increase in TG. Liver histopathology was similar in all groups and within the normal range. CONCLUSION: A moderate amount of Alc daily together with S is safe in rats. Additionally, S administration in Wistar rats diminishes the Alc-induced TG and AST rises. PMID- 14758717 TI - Metals and metal compounds in carcinogenesis. AB - Several metals and metal containing compounds are potent mutagens and carcinogens. The most often blamed are chromium, arsenic, nickel, vanadium, iron, copper and manganese. Although each of them has its own mechanism of action, it is believed that most of their mechanisms of action involve reactive oxygen species (ROS). Furthermore, nickel modulates gene expression by induction of DNA methylation and/or suppression of histone acetylation. Arsenic activity on cell metabolism is multiple; it seems that cell transformation is induced by long-term exposure to a low level of arsenic. The paradox of arsenic is that it has also a valuable therapeutic efficacy in cancer treatment. Manganese is known to cause DNA damage, although it does not represent a significant carcinogenic risk. Magnesium deficiency and iron excess are not exactly carcinogenetic, but certain concentrations of these metal ions are needed to prevent cancer. PMID- 14758718 TI - Extracts of spice and food plants from Thai traditional medicine inhibit the growth of the human carcinogen Helicobacter pylori. AB - BACKGROUND: Helicobacter pylori (HP) is a gramnegative bacterium and well recognized as being the primary etiological agent responsible for the development of gastritis, dyspepsia, peptic ulcer disease and gastric cancer. In developing countries, a high prevalence of HP infection is associated with an increased incidence of gastric cancer. Thailand, however, while having a high prevalence of HP infections, has a lower than expected gastric cancer rate than other developing countries. It has been suggested that the diet and life style in Thailand may explain this discrepancy. MATERIALS AND METHODS: The in vitro susceptibility of 18 strains of HP to 20 extracts of spice and food plants used in Thai traditional medicine for the treatment of GI disorders was assessed. RESULTS: Methanol extracts of Myristica fragrans (aril) inhibited the growth of all HP strains with minimum inhibitory concentration (MIC) of 12.5 micrograms/ml; extracts from Barringtonia acutangula (leaf) and Kaempferia galanga (rhizome) had an MIC of 25.0 micrograms/ml; Cassia grandis (leaf), Cleome viscosa (leaf), Myristica fragrans (leaf) and Syzygium aromaticum (leaf) had MICs of 50.0 micrograms/ml. Extracts with an MIC of 100.0 micrograms/ml included Pouzolzia pentandra (leaf), Cycas siamensis (leaf), Litsea elliptica (leaf) and Melaleuca quinquenervia (leaf). CONCLUSION: Plants used in Thai traditional medicine to treat gastrointestinal ailments inhibit the growth of HP. These data indicate that these plants may have chemopreventative activities and thus may partly explain the reduced incidence of gastric cancer in Thailand. PMID- 14758719 TI - Inhibitory effects of autoimmune disease by green tea in MRL-Faslprcg/Faslprcg mice. AB - To investigate whether green tea has inhibitory effects on the development of autoimmune disease (AID), one-month-old MRL-Faslprcg/Faslprcg mice were fed diets containing 2% green tea powder (GTP) for 3 months. At the end of GTP feeding, the weights of body, subcutaneous (s.c.) and intraperitoneal (i.p.) lymph nodes (LN), kidneys, spleen and intraperitoneal adipose tissue (IPAT), serological abnormalities and renal lesions were compared between GTP-fed and control mice. SCLN, IPLN, kidneys and IPAT weights in both sexes, spleen weight in males and body weight increase in males were significantly lower in GTP-fed mice. Particularly, LN hyperplasia and fatty accumulation were markedly reduced by GTP. Serum levels of anti-DNA antibodies and immune complexes (IC) were significantly lowered and proteinuria and blood urea nitrogen tended to be improved by GTP. The incidence of serious glomerulonephritis was significantly lower and nephric vasculitis was almost completely prevented in GTP-fed mice. Moreover, the survival of mice was significantly prolonged by GTP feeding for 6 months. These results indicate that the progression of lupus-like syndrome including glomerulonephritis was significantly delayed by reduced production of autoantibodies and IC in GTP-fed MRL-Faslprcg/Faslprcg mice, which led to the prolonged survival. PMID- 14758720 TI - Induction of apoptosis by Citrus paradisi essential oil in human leukemic (HL-60) cells. AB - Limonene is a primary component of citrus essential oils (EOs) and has been reported to induce apoptosis on tumor cells. Little is known about induction of apoptosis by citrus EOs. In this study, we examined induction of apoptosis by Citrus aurantium var. dulcis (sweet orange) EO, Citrus paradisi (grapefruit) EO and Citrus limon (lemon) EO. These EOs induced apoptosis in HL-60 cells and the apoptosis activities were related to the limonene content of the EOs. Moreover, sweet orange EO and grapefruit EO may contain components besides limonene that have apoptotic activity. To identify the components with apoptotic activity, grapefruit EO was fractionated using silica gel columns, and the components were analyzed by GC-MS. The n-hexane fraction contained limonene, and the dichloromethane fraction (DF) contained aldehyde compounds and nootkatone. Decanal, octanal and citral in the DF showed strong apoptotic activity, suggesting that the aldehyde compounds induced apoptosis strongly in HL-60 cells. PMID- 14758721 TI - Up-regulation of thymosin beta 4 gene expression in experimentally-induced uterine adenomyosis in mice. AB - Degradation of the myometrium contributes to the development of adenomyosis and disruption of the actin cytoskeleton occurs in the myometrium adjacent to the invasive endometrium. A subtractive cDNA library that contained genes up regulated in the uterus with adenomyosis induced by pituitary grafting was constructed in mice. Thymosin beta 4 (Tb4), an actin sequestering protein, was contained in the library and was confirmed to be abundant in the adenomyotic uteri. Tb4-immunoreactivity was shown in the myometrium, with a stronger signal being observed in the area closer to the tip of endometrial tissue that is invading the myometrium. Up-regulation of Tb4 gene expression may be responsible for the disintegration of the musculature during the development of adenomyosis. PMID- 14758722 TI - Serotonin activity and liver dysfunction following hepatic ischemia and reperfusion. AB - Serotonin (5-hydroxytriptamine; 5-HT), which is stored in platelets, is known to induce vasoconstriction and promote platelet aggregation. More recent studies suggest that serotonin also plays a role in organ injury after ischemia and reperfusion. The purpose of this study was to characterize the role of 5-HT and platelet function in the pathogenesis of hepatic ischemia-reperfusion injury. Under the portocaval shunt, 60 or 90 min of complete warm ischemia of canine liver was induced by Pringle's maneuver, followed by reperfusion for 120 min. Time-matched, sham-operated animals served as controls. Hepatic tissue blood flow and various parameters of hepatic vein blood (ALT, LDH, platelet count and platelet aggregation) were measured before and after reperfusion. 5-HT levels in portal vein and hepatic vein were also assayed. Hepatic ischemia and reperfusion resulted in liver hypoperfusion, hepatocellular dysfunction, increased platelet aggregation, increased 5-HT levels, and hepatic microcirculation injury. These results suggest that the endogenous 5-HT released from platelet may contribute to liver tissue hypoperfusion following hepatic ischemia-reperfusion. PMID- 14758723 TI - The usefulness of p63 as a marker of breast myoepithelial cells. AB - The identification of an outer layer of myoepithelial cells is a valuable clue in the differential diagnosis of breast lesions. Myoepithelial cells can usually be appreciated with standard hematoxylin-eosin stains, however in pathology practice one encounters difficult cases, particularly in core biopsy specimens. There are several reported markers for the immunohistochemical detection of myoepithelial cells. Smooth muscle specific proteins, such as smooth muscle actin, smooth muscle myosin heavy chain, calponin and h-caldesmone are utilized to highlight myoepithelium. Smooth muscle actin is the most commonly used and has been established as a specific and sensitive marker. However, sometimes the staining can not be interpreted, since actin stains stromal fibroblasts and vascular smooth muscle cells as well. S100 protein and specific cytokeratins (keratins 5,7,14 and 17) also stain myoepithelial cells, but the staining is not specific and is not optimally sensitive. Maspin and CD10 are relatively new and promising markers. The nuclear protein p63 has attracted much attention in recent reports. p63-positive myoepithelial cells have been shown to surround benign epithelial lesions and form a consistent, although discontinuous, rim around epithelial cells in carcinomas in situ. No staining has been noted in infiltrative carcinomas. p63-immunostaining is nuclear and so it is easily appreciated, even in cytologic preparations. It is also highly specific since neither stromal fibroblasts nor vascular smooth muscle cells are stained. In conclusion, it appears that p63 is a sensitive and specific myoepithelial marker and may be included in immunohistochemical panels aiming at identifying myoepithelial cells in problematic breast lesions. PMID- 14758724 TI - Comparison of cytotoxicity and radical scavenging activity between tea extracts and Chinese medicines. AB - Three hot water extracts of black tea, green tea and powdered green tea and five Chinese medicines (Shosaiko-tou, Orengedoku-tou, Goshuyu-tou, Choto-san, Keishininjinn-tou) were investigated for their ability to modify nitric oxide (NO) production by lipopolysaccharide (LPS)-stimulated mouse macrophage-like Raw 264.7 cells, and for their cytotoxicity, radical intensity and scavenging activity. All eight materials significantly reduced the extracellular concentration of NO in the LPS-stimulated Raw 264.7 cells. ESR spectroscopy shows that tea extracts, which had higher cytotoxicity, generated higher amounts of radicals, and more efficiently scavenged O2- (generated by hypoxanthine-xanthine oxidase reaction), hydroxyl radical (generated by Fenton reaction) and NO (generated by 1-hydroxyl-2-oxo-3-(N-3-methyl-3-aminopropyl)-3-methyl-1-triazene) than Chinese medicines. Close association between the radical intensity and radical scavenging activity suggests their bimodal (anti-oxidant and pro-oxidant) action. Pretreatment of mice with tea extracts significantly reduced the lethality of Escherichia coli-infection. All tea extracts showed no apparent anti HIV activity. The present study demonstrates, for the first time, several attractive features of tea extracts in comparison with Chinese medicines, suggesting the possible application of the tea extracts for radical-mediated diseases. PMID- 14758725 TI - Effect of endodontic agents on cytotoxicity induction by sodium fluoride. AB - We investigated six endodontic agents for their ability to induce apoptosis and modify the cytotoxic activity of NaF against human squamous cell carcinoma (HSC 2) and human promyelocytic leukemia (HL-60) cell lines. Four Group I agents (Form Cresol, Cam Phenic, Eucaly Soft, GC Fuji Varnish), but not two Group II agents (Caviton, Canals-N), induced internucleosomal DNA fragmentation and activated caspases 3, 8 and 9 in HL-60 cells. Only Cam Phenic among these agents additively enhanced the cytotoxic activity of NaF in HSC-2 and HL-60 cells. Form Cresol and Cam Phenic reduced the glucose consumption at early stage, possibly due to their toxic effect. Amino acid analysis suggests that the higher cytotoxicity of Form Cresol may be derived, at least in part, from its oxidizing action. PMID- 14758726 TI - Circadian periodicity of plasma lipid peroxides and other anti-oxidants as putative markers in gynecological malignancies. AB - BACKGROUND: The chronome (from chronos, time, and nomos, rule), or time structure, of lipid peroxidation and anti-oxidant defense mechanisms may relate to prevention and curative chronochemotherapeutic efficacy and management. PATIENTS AND METHODS: Newly diagnosed women with gynecological malignancies (N = 30), 30-60 years of age, and age-matched clinically healthy women (N = 35) provided blood samples every 6 hours for 24 hours under standardized conditions. Plasma malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and glutathione reductase (GR) activities, and serum ascorbate, urate and high-density lipoprotein cholesterol (HDL-C) concentrations were determined. RESULTS: Each variable underwent circadian variation (p < or = 0.002). Patients differed from controls by their overall chronome-adjusted mean value (MESOR) and by the circadian dynamics in the spectral element of their chronome. CONCLUSION: Chronomes of putative anti- and pro-oxidants should be mapped to explore their putative chemotherapeutic role as markers in cancer chronoprevention and management of established disease. PMID- 14758727 TI - Subcellular fractionation and electrophoretic analysis of proteins from the regenerating nerve in rabbits following treatment with triiodothyronine (T3). AB - BACKGROUND: Triidothyronine(T3) plays a critical role in the physiological function of virtually all tissues. In the present study the effect of T3 on nerve regeneration was studied. MATERIALS AND METHODS: The right sciatic nerve of 20 male rabbits was axonotomised. Ten of the animals were used as controls while the others received T3. Seven days later, all were sacrificed. Both sciatic nerves were excised and each axonotomised nerve was divided into three equal segments: I (proximal to the lesion), II (containing the lesion), III (distal to the lesion). Corresponding segments from the contralateral nerves were used as controls. After subcellular fractionation, proteins were analyzed by polyacrylamide gel electrophoresis and identified by proteomic analysis. RESULTS: T3 administration increased the levels of tubulins alpha 6 and beta 3, beta-actin, myelin protein zero, myelin basic protein, myelin protein-2 and histones H3 and H2A in segments I and III of the injured nerves. Increases in the levels of beta-actin and Y box binding protein correlated to the event of injury rather than T3 administration. CONCLUSION: Our findings indicate that T3 treatment positively affects peripheral nerve regeneration. PMID- 14758728 TI - Tumor markers. An update approach for their prognostic significance. Part I. AB - Identification of prognostic tumor markers is a main strategy for planning treatment and predicting outcome of patients with various malignancies. This article reviews a panel of proliferation markers, hormone receptors, oncogene products and apoptosis regulators that have shown potential as prognostic indicators in common human cancers. At present, steroid receptors and the c-erbB 2 gene are the only tissue-based markers accepted in clinical practice, having an established role in breast cancer prognosis. Other markers e.g., the p53 gene, look promising as prognostic factors in different kinds of cancer. PMID- 14758729 TI - Investigation about origin and spreading of neoendothelium in autologous arterial vessel replacement. AB - INTRODUCTION: In many fields of surgery an autologous arterial vessel replacement is used. Because of trauma and insufficient nutrition, a loss of complete endothelium in the replaced vessel segment and at the anastomoses can be observed within a few hours. The origin and the spread of the neoendothelium are unknown. Our study uses Von-Willebrand-protein, a product of endothelium cells, as a marker to obtain new information about the origin and spread of the new endothelial cells. MATERIALS AND METHODS: Seventy Wistar rats, seven groups of 10 animals each, were operated. A four-millimetre-long segment of the common carotid artery was isolated and reinserted. After a varying length of time (between directly after the operation and six months after), the common carotid artery including bifurcation was isolated after cardioperfusion. Carotid arteries were embedded and cross-sections were stained with hematoxylineosin, Verhoeff's tissue elastin stain and immunohistochemical anti-Von-Willebrand-factor-antibody. The complete vessel was divided into nine points of measurement on each side, three points at each anastomosis and three points in transplanted segment. There the number of positive endothelial cells was rated. RESULTS: Twenty-four hours after the operation no further endothelium was detectable in autologous transplant. Stainings eight days after operation contained Von-Willebrand-factor-positive cells in luminal cell layers. Near to the anastomosis excessive myointimal hyperplasia was detectable. Also, eight days after operation, some positive endothelial cells in adventitia were seen near to the anastomosis in small vessel lumina. Four weeks after operation, the luminal endothelium was completely regenerated and the luminal endothelial layer was confluent. Eight days after the operation regeneration in lateral regions was faster than in the region of the transplant. CONCLUSION: In our experiment regeneration in lateral regions was faster than in the region of the transplant. Early staining in adventitia and proof of newly formed vasa vasora in adventitia may be a sign of a possible migration from adventitia to luminal area. PMID- 14758730 TI - GST M1 and CYP1A1 gene polymorphism and daily fruit consumption in Turkish patients with non-small cell lung carcinomas. AB - BACKGROUND: In general, the metabolism of carcinogens involves two pathways. The oxidative pathway, which enhances carcinogenesis (phase I), and the protective pathway, in which carcinogens are conjugated with a series of substances such as glutathione to achieve detoxification (phase II). It has been suggested that an increased phase I enzyme activity (CYP1A1) and a decreased phase II enzyme activity (GST M1) could each individually cause an increase in the risk of cancer. MATERIALS AND METHODS: In the present study we explored the association between genetic polymorphisms of CYP 1A1 and GST M1 and non-small cell lung cancer (n = 55) and controls(n = 60) in Turkish subjects. We used PCR methods and enzyme restriction for determining polymorphism. A standard food questionnaire was used to determine daily fresh fruit consumption. RESULTS AND CONCLUSION: We found that CYP1A1 mutant variant (Ile/Val) was more highly expressed in Turkish patients and controls than in other Caucasian populations. Our findings were similar to Far Eastern populations (32.7% for patient group, 43.1% for controls). Inspite of the similarity between the groups regarding GST M1 polymorphism, in the patient group, patients with GST M1 null genotype had a statistically significant positive history of exposure to carcinogens other than smoking, such as asbestos, petrochemicals and/or other chemicals (p = 0.01). The patients, who had CYP 1A1 mutant variant, had increased risk of adenocarcinoma (p = 0.046) of lung (8 out of 18 patients) and 6 of them also had GST M1 (-) gene variants together. The patients who consumed less fruit daily had a greater risk of epidermoid carcinoma of lung (p = 0.019). However this study showed that there were no differences between the patient and control groups regarding genetic polymorphism of genes. PMID- 14758731 TI - CD44 family and gynaecological cancer. AB - CD44 refers to a multifunctional family of type I transmembrane proteins. The CD44 gene contains at least 21 exons, 11 of which can be variably spliced and produce a variety of heavily glycosylated cell surface proteins, known as CD44 variant isoforms. These proteins have been implicated in many biological processes, such as cell adhesion, cell substrate, cell to cell interactions, including lymphocyte homing haemopoiesis, cell migration and metastasis. These abilities are of great importance in chronic inflammation and in cancer. Published data have shown that CD44 has the ability to recruit leucocytes to vascular endothelium at sites of inflammation, which is one of the first steps in the inflammatory response. In cancer, deregulation of the adhesion mechanisms increases the ability of tumor cells to metastasis. This behavior seems to be explained by the existing relationship between hyaluronan, a basic component of the extracellular matrix and CD44, which is its major cell surface receptor. There are CD44 variant isoforms which are expressed on different types of normal cells. In addition some isoforms are overexpressed on tumor cells including breast, cervical, endometrial and ovarian cancer. This property seems to be correlated with the metastatic potential of these cells. This review summarizes the available data on the possible prognostic role of the polymorphic CD44 protein family and its role as a tumor marker in gynaecological cancer. PMID- 14758732 TI - Effects of 6-gingerol, an antioxidant from ginger, on inducing apoptosis in human leukemic HL-60 cells. AB - 6-Gingerol, a naturally occurring plant phenol, is one of the major components of fresh ginger. In this paper, the antioxidative effects of 6-gingerol were detected by DPPH and DCFH assays and, as predicted, 6-gingerol as an antioxidant was shown to protect HL-60 cells from oxidative stress. Moreover, it induced cell death in promyelocytic leukemia HL-60 cells, caused DNA fragmentation and inhibited Bcl-2 expression in HL-60 cells. These results suggested that the inhibition of Bcl-2 expression in HL-60 cells might account for the mechanism of 6-gingerol-induced apoptosis. In the inhibitory assay, the cytotoxic effect of 6 gingerol could be prevented by catalase. We suggest that 6-gingerol induced cell death by mediating reactive oxygen species such as hydrogen peroxide and the superoxide anion. Therefore, the results showed that 6-gingerol induced apoptosis in HL-60 cells, not due to its antioxidative activity. PMID- 14758733 TI - A psychoneuroendocrine study of brain dopaminergic sensitivity in locally limited or metastatic cancer patients. AB - In addition to the occurrence of pain, the evidence of a diminished capacity to feel pleasure is one of the most common cancer-related symptoms. Recent advances in psychoneuroendocrinological knowledge has shown that the perception of pleasure is mainly mediated by the dopaminergic pathways in the brain. Moreover, it has also been demonstrated that the brain dopaminergic sensitivity may be clinically explored by evaluating the endocrine response to the administration of dopaminergic agents, such as apomorphine, which consists of a decline in PRL concentrations and an increase in GH and cortisol levels. The present study was performed to evaluate dopaminergic sensitivity by the administration of apomorphine in cancer patients in an attempt to document possible cancer-related neuroendocrine anomalies, which could explain the psychological status of the patients. The study included 24 cancer patients (breast cancer: 12; colorectal cancer: 7; non-small cell lung cancer: 5), 12 of whom showed distant organ metastases. Apomorphine was given orally at 0.01 mg/kg b.w., by collecting venous blood samples before and after 20 and 60 minutes. A normal decline in PRL levels was seen in both non-metastatic and metastatic cancer patients. No cortisol increase in response to apomorphine was achieved and the lack of cortisol response was particularly evident in metastatic patients. No GH rise occurred in either metastatic or non-metastatic cancer patients. Finally, no significant difference in the endocrine response to apomorphine was seen in relation to the histotype of tumor. The results of this study show that the neoplastic disease is characterized by neurochemical alterations involving pleasure-related dopaminergic pathways, which are more evident in the metastatic disease, without particular differences in relation to tumor histotype. Therefore, the psychological condition of cancer patients would not depend only on psychological factors, but it could be due at least in part to cancer-related neuroendocrine alterations involving the dopaminergic system. PMID- 14758734 TI - Cystitis glanduralis complicating an eosinophilic cystitis: a case report. AB - We report on a rare case of cystitis glandularis complicating an eosinophilic cystitis in an adult. Complaints at presentation included dysuria, haematuria and abdominal pain. Ultrasound and cystoscopy suggested a bladder tumor. Histological analysis of bladder biopsy showed the typical findings of cystitis glandularis associated with eosinophilic cystitis. The patient was treated with transurethral resection of the lesion and a combination of corticosteroids and anthistaminics for three months. He is disease-free at 24 months of follow-up. PMID- 14758735 TI - GnRH agonist administration after embryo transfer, in long protocol stimulated cycles, prevents ovarian hyperstimulation syndrome. A report of five cases. AB - BACKGROUND: Ovarian hyperstimulation syndrome (OHSS) is a frequent serious complication in controlled ovarian hyperstimulation for in vitro fertilization. Among the treatments for OHSS, the continuation of GnRH agonist during the luteal phase has been recently suggested. MATERIALS AND METHODS: Five patients at risk for OHSS received 0.1 mg triptorelin for one week after embryo transfer in order to prevent OHSS. RESULTS: All the five patients developed mild symptoms. None of them required hospitalization. CONCLUSION: Although the pathophysiology of OHSS and the effects of GnRH agonist during the luteal phase remain unclear, the continuation of GnRH agonist seems to be a safe and effective treatment for OHSS. PMID- 14758736 TI - Cost hikes, direct contracts spur capitation rebound. PMID- 14758737 TI - Blue Cross of California implements strategies to boost capitation, quality. PMID- 14758738 TI - Financial survey shows capitation revenue increasing, but medical group profits are sliding. PMID- 14758739 TI - Demand management: not in vogue, but working well in practice. PMID- 14758740 TI - California data sharing project shows promise, report finds. PMID- 14758741 TI - Linking organization of work and health: the challenge. PMID- 14758742 TI - Work organization: the neglected child of (social) epidemiology. PMID- 14758743 TI - Comment on the obesity issue. PMID- 14758744 TI - Obesity: where is the next focus? PMID- 14758745 TI - Fighting the obesity epidemic. PMID- 14758746 TI - Monitoring the changing organization of work: international practices and new developments in the United States. AB - Recent trends in the organization of work have raised concerns about their implications for safety and health in the workplace. Capacity for monitoring of these trends from an occupational safety and health perspective (also known as hazard surveillance) varies considerably across countries and regions. This forum article discusses current practices for monitoring the organization of work, noting strengths, limitations, and needs for improvement. Particular attention is given to the status of monitoring practices in the U.S., and new initiatives by the National Institute for Occupational Safety and Health (NIOSH) to improve upon these practices. PMID- 14758747 TI - [Predictive validity of a brief scale to assess subjective prognosis of work capacity (SPE Scale) in a cohort of LVA insured patients with severe back pain or functional complaints relating to internal medicine]. AB - OBJECTIVES: Vocational (dis-)ability is a central concept in social medicine. Beside from medical factors psychological and system factors play a crucial role in determining vocational disability. The development of instruments assessing the prognosis of gainful employment is an important task in rehabilitation research. METHODS: A short scale measuring the subjective prognosis of gainful employment (SPE-scale) was administered to 481 blue collar workers suffering from severe back pain or else functional syndromes. Employment status two or three years following the assessment is known. RESULTS: 11% of the subjects have applied for early retirement; 5% actually have retired. There are statistically significant relations of SPE-scores and subjects employment status. Effect sizes of the SPE-scores equal or even outrank those of other instruments assessing vocational (dis-)ability. The positive predictive power of the scale is low, though. CONCLUSIONS: The SPE scale can be recommended for further use in (rehabilitation) research. The employment of the instrument for medical appraisal has to be viewed with more caution. The percentage of subjects at risk for early retirement is largely overestimated in case of a positive test. Instruments like the SPE-scale can assist but never replace the medical appraisal. PMID- 14758748 TI - [Return to work after cardiologic rehabilitation]. AB - OBJECTIVES: The objectives of the present study were to determine prospectively return to work and its predictors in patients after cardiac rehabilitation. METHODS: Patients were enrolled at admission to inpatient cardiac rehabilitation centres (n = 18). Primary indications for admission were myocardial infarction, coronary artery bypass grafting or percutaneous transluminal coronary angioplasty. RESULTS: We included 2441 consecutive patients (1907 men, mean age: 60 +/- 10 years; 534 women, mean age: 65 +/- 10 years). A total of 43% of all patients had been actively employed before the event. Of these patients, 65% had returned to work six months and 67% 12 months after cardiac rehabilitation. Successful return to work after 12 months was significantly predicted by younger age, non-manual work, self-employment, a higher physical and mental quality of life, and a better exercise ECG result. CONCLUSION: Return to work is predicted by sociodemographic factors, quality of life, and the exercise ECG at the rehabilitation centre. The determination of early predictors for return to work may aid to identify patients particularly at risk for failure to return to work. PMID- 14758749 TI - Determinants of health policy impact: comparative results of a European policymaker study. AB - OBJECTIVES: This article will use a new theoretical framework for the analysis of health policy impact introduced by Rutten et al. (2003). In particular, it will report on a comparative European study of policymakers' perception and evaluation of specific determinants of the policy impact, both in terms of output (implemented measures) and outcome (health behaviour change). Policy determinants investigated are goals, resources, obligations and opportunities as related to the policymaking process. METHODS: Theory is applied to a comparative analysis of prevention and health promotion policy in Belgium, Finland, Germany. The Netherlands, Spain and Switzerland. The study is MED2-part of a project that has developed a Methodology for the Analysis of the Rationality and Effectiveness of Prevention and Health Promotion Strategies (MAREPS) within the EU-BIO-program. A mail survey of 719 policymakers on the executive and administrative level selected by a focused sample procedure was conducted. This survey used policymakers' experience and evaluative expertise to analyse determinants of policy output and outcome. RESULTS: Regression analyses reveal differential predictive power of policy goals, resources, obligations, as well as of political, organisational and public opportunities. For instance, whereas resources, concreteness of goals, and public opportunities have significant importance for health outcome of policy, obligations and organisational opportunities significantly predict policy output. CONCLUSIONS: Results are discussed in terms of rationality and effectiveness of health policy. They indicate that six sensitising constructs derived from the theoretical framework represent equivalent structures across nations. They comprise a validated instrument that can be used for further comparative health policy research. PMID- 14758750 TI - Testing questionnaires with cognitive methods: Part 2, Thinkalouds. PMID- 14758751 TI - Web alert. The chemistry of process development. PMID- 14758752 TI - The Pictet-Spengler reaction in solid-phase combinatorial chemistry. AB - The Pictet-Spengler reaction is an important reaction for the generation of tetrahydro-beta-carbolines and tetrahydroisoquinoline ring systems, which exhibit a range of biological and pharmacological properties. This review covers the solid-phase Pictet-Spengler reaction, as employed in solid-phase routes toward a range of complex heterocyclic ring systems, with focus on experimental conditions, efficiency and diastereoselectivity. This is illustrated by the application of this reaction to the synthesis of combinatorial libraries, natural product analogs and drug-like scaffolds. PMID- 14758753 TI - Solid-phase strategies for the design and synthesis of heterocyclic molecules of medicinal interest. AB - Recently, there has been a wealth of reports concerning the polymer-supported traceless synthesis of heterocyclic structures of medicinal interest. In 2002, important advances included the continued use of cyclative cleavage strategies to obtain novel heterocyclic structures with high chemical diversity. In addition to these strategies, several reports based on a novel approach involving the use of common combinatorial scaffolds for the generation of a variety of heterocyclic structures have also appeared. This review covers the application of these two approaches to a wide array of compounds based on azoles quinazolines, indoles, pyridines, isoxazolines, piperazines and pyrroles. PMID- 14758754 TI - Solid-phase synthesis of deglycobleomycin and bleomycin. AB - The bleomycins are a group of structurally related glycopeptide antibiotics originally isolated from Streptomyces verticillus in the 1960s. They are used for the treatment of cancers including Hodgkin's lymphomas, carcinomas of the skin, head and neck, and testicular tumors. Whereas analogs of bleomycin have been synthesized to facilitate an understanding of its biochemical properties, the complexity of the molecule has severely limited the total number of analogs synthesized. The solid-phase synthesis of fully functional and active deglycobleomycin and bleomycin analogs is described. This approach has resulted in the synthesis of over 160 unique deglycobleomycin and bleomycin analogs in the past two years, an accomplishment that would not have been possible without the solid-phase methodology. PMID- 14758755 TI - Searching for scalable processes: addressing the challenges in times of increasing complexity. AB - The position of the pharmaceutical industry at the beginning of the 21st Century and its role in process research and development are important considerations. Based on recent figures and statistics, it is evident that new and maybe unorthodox ways of operating need to be envisaged to counteract the explosive rise in research and development costs (> US $800 million per successful launch in 2002) and the worryingly high level of attrition (approximately 90%), which, in addition to long handling times for authority approval, lead to a continued decline in the number of drugs entering the market. In addition to pointing out these harsh realities, it is essential to remember that the time taken to bring a new product forward is crucial. In this context, a front-loading model has been developed and applied in AstraZeneca's process research and development. The enormous implications of chirality in the pharmacological arena have been fully acknowledged for many years, and sales figures of chiral molecules corroborate the importance of this feature. The methodologies that are available to address the design and scale-up of production methods are also reviewed against a backdrop of recent in-house cases. PMID- 14758756 TI - Protein crystallization: from HTS to kilogram-scale. AB - The first experiments on protein crystallization started randomly during the 19th century. This technique has been widely used for the determination of the tertiary structure of proteins since the 1950s, when an understanding of the physics of protein crystallization began to emerge. In the 1980s and 1990s, research focused on the study of protein crystal growth processes in microgravity environments, which were created in space shuttle experiments. High-throughput screening (HTS) systems were developed that later found broader laboratory applications. The combination of HTS with an engineering approach opens new opportunities for the protein crystallization process to become a robust, scalable, reproducible and economically viable industrial unit operation. PMID- 14758757 TI - Asymmetric hydrogenation of pharmaceutically interesting substrates. AB - The application of asymmetric hydrogenation to the manufacture of pharmaceutical intermediates has become increasingly prevalent in the past three to five years. This review focuses on the hydrogenation substrates and their use, rather than comparing the specific catalysts employed. The literature published between 2002 and 2003 is covered, with some pertinent examples from 2001, all of which demonstrate the increasing number of applications for this technology. PMID- 14758758 TI - Synthetic applications of palladium-catalyzed hydroarylation and related systems. AB - Synthetic organic chemists have recognized the hydroarylation of alkynes and alkenes as a versatile methodology. In the first part of this review, the regio- and stereochemistry of the hydroarylation of alkynes and the synthetic applications of this reaction in the synthesis of antiviral agents and steroid derivatives will be discussed. In the second part, applications of the hydroarylation of alkenes in the syntheses of epibatidine analogs, argemonine, retinoid X receptor (RXR), retinoic acid receptor (RAR) modulators and neurokinin (NK) receptor NK1 antagonists will be discussed. Emphasis will be placed on the application of Pd-catalyzed hydroarylation to the synthesis of biologically active compounds. PMID- 14758759 TI - Recent progress in enzymatic resolution and desymmetrization of pharmaceuticals and their intermediates. AB - The literature since August 2000 is surveyed for interesting and useful examples of the resolution or desymmetrization of pharmaceutical entities, their intermediates and potential drug precursors. Existing, putative and developmental drugs used in various therapeutic areas are discussed. PMID- 14758760 TI - Microbial/enzymatic synthesis of chiral pharmaceutical intermediates. AB - Chirality is a key factor in the efficacy of many drugs; thus, the production of single enantiomers of drug intermediates has become increasingly important in the pharmaceutical industry. Chiral intermediates and fine chemicals are in high demand from the pharmaceutical and agrochemical industries for the preparation of bulk drug substances and agricultural products. There has been an increasing awareness of the enormous potential of microorganisms and enzymes for the transformation of synthetic chemicals with high chemo-, regio- and enantioselectivity. In this article, biocatalytic processes for the synthesis of chiral pharmaceutical intermediates are described. PMID- 14758761 TI - Rapid assembly of molecular diversity via exploitation of isocyanide-based multi component reactions. AB - Molecular diversity is the variability of physical properties between molecules, viewed in terms of molecular shape, polarity/charge, lipophilicity, polarizability and flexibility. Due to their widespread medicinal properties, natural products were one of the original sources of molecular diversity; however, new developments in the search for novel pharmacological agents over the last decade have focused on the preparation of chemical libraries as the source of new leads for drug discovery. A plethora of personal synthesizers and new automation technologies have emerged to help fuel the lead discovery engines of drug discovery organizations. Multistep solid-phase syntheses of diverse libraries in excess of 10,000 products can now be prepared via split-and-mix techniques. Simultaneously, a multitude of more efficient, diversity- or target oriented solution-phase chemical methodologies have appeared in the chemical literature, enabling the relatively facile construction of successful lead generation libraries with low full-time equivalent input and little capital expenditure. Isocyanide-related multi-component reactions hold a pre-eminent position in this regard, and are finding increasing applications in the discovery process of new drugs and agrochemicals. This review is the authors' personal assessment of advances in the field over the last two years (2002 to 2003), with little emphasis placed on highly mechanistic details. PMID- 14758762 TI - Progress toward the synthesis of hinckdentine A. AB - Hinckdentine A is a structurally unique marine alkaloid containing an 11b,12,13,14,15,16-hexahydroazepino[4',5':2,3] indolo[1,2-c]quinazoline ring system. This review covers the literature on hinckdentine A. The indolo[1,2 c]quinazoline ring system is the foundation of the hinckdentine A structure. The literature on the chemistry of indoloquinazolines until the end of 1994 has been reviewed by Billimoria and Cava. Therefore, we review the research carried out on the synthesis of indolo[1,2-c]quinazolines and related ring systems in which hinckdentine A is described from 1995 to present. The construction of the quaternary center is the primary synthetic challenge posed by the hinckdentine A structure. We will present a selection of methods applicable to the synthesis of 2,2-disubstituted indolines. Finally, we present the three most significant attempts to synthesize hinckdentine A. PMID- 14758763 TI - Synthetic methodology utilized to prepare substituted imidazole p38 MAP kinase inhibitors. AB - In this manuscript, we review the synthetic methods utilized to prepare a variety of imidazole p38 MAP kinase inhibitors. Several methods have been used to prepare the key templates that are discussed; manipulation of the heterocycles around the imidazole core are also considered in the context of their biological activity. Finally, we discuss new synthetic methodologies discovered in our laboratories, which are useful for the preparation of a member of this class of tetrasubstituted imidazole p38 inhibitors. The optimal route involves the thiazolium-catalyzed cross-benzoin condensation of a pyridine aldehyde with an N acylimine. The pyridine aldehyde was prepared in three steps and 68% yield from 2 chloro-4-cyanopyridine. The tosylamide precursor to the N-acylimine was prepared in two steps and 93% yield from isonipecotic acid. We demonstrate the scope and some preliminary mechanistic studies concerning this new reaction. The resulting alpha-ketoamide is then cyclized with methyl ammonium acetate to provide the desired tetrasubstituted imidazole. Cbz deprotection and formation of a pharmaceutically acceptable salt completes the synthesis in six steps and 38% overall yield. PMID- 14758764 TI - Advances in multivariate analysis in pharmaceutical process development. AB - In the last five years, reports of the application of multivariate methods in pharmaceutical process research and development have burgeoned. Examples range from the widespread adoption of statistical experimental design for screening and process optimization to the implementation of workflows that integrate multivariate characterization, experimental design and the development of quantitative structure-property relationships. Having learned from the application of these techniques in drug discovery and armed with modern high throughput experimentation platforms, practitioners are discovering the power of experimental design and multivariate methods for shortening process development timelines. PMID- 14758765 TI - T-cell depletion for transplant tolerance induction: promises and hurdles. PMID- 14758766 TI - The Th1 immune pathway as a therapeutic target in Crohn's disease. AB - Crohn's disease (CD) is a chronic inflammatory condition of the gastrointestinal tract, characterized by overactive T-helper (Th)1-mediated responses towards resident bacterial flora in genetically susceptible individuals. Biotechnological methodology has allowed the development of therapeutic compounds that specifically target single, well-defined pathways, thereby inhibiting abnormal Th1 polarization. These therapies include antibody-based, as well as nucleic-acid related approaches that target various pro-inflammatory cytokines, as well as intracellular signaling pathways. This review summarizes treatments already in clinical use and discusses novel targets that offer additional therapeutic opportunities in the management of CD. PMID- 14758767 TI - Transplantation: toxicokinetics and mechanisms of toxicity of cyclosporine and macrolides. AB - For over two decades, calcineurin inhibitors (CIs) have been the mainstay of immunosuppressive therapy following solid-organ transplantation. However, CI nephrotoxicity is one of the main contributors to chronic kidney allograft dysfunction. A novel class of immunosuppressants that inhibit the kinase mammalian target of rapamycin (mTOR), although not nephrotoxic themselves, enhance CI nephrotoxicity. The biochemical basis of CI toxicity and their toxicodynamic interaction with mTOR inhibitors is still poorly understood. Studies using a magnetic resonance spectroscopy-based metabonomic approach indicate that CI toxicity is caused by drug-induced mitochondrial dysfunction and that mTOR inhibitors enhance the negative effects of CIs on cell energy metabolism. PMID- 14758768 TI - JAK3 inhibition as a new concept for immune suppression. AB - Although current immunosuppressive drugs are effective, they have numerous severe side effects that mandate the search for new agents. Mutations in the gene for janus kinase (JAK)3 result in severe combined immune deficiency with severely impaired humoral and cellular immunity, an observation that has prompted the development of JAK3 inhibitors. Due to its central role in lymphocyte activation, proliferation and homeostasis, targeting the JAK/signal transducer and activator of transcription (STAT) pathway may provide the required efficacy, without the toxicities associated with current therapies. Several studies conducted in rodents have validated the proof-of-concept, with a variety of JAK3 inhibitors demonstrating efficacy for immune suppression. In addition, the selective JAK3 inhibitor CP-690550 (Pfizer Inc) significantly improved allograft survival in a stringent preclinical model in primates and exhibited a good safety profile in non-human primates. This, along with studies of protein kinase inhibitors for cancer treatment, could demonstrate that development of effective, safe and selective kinase inhibitors for immunosuppression is possible. PMID- 14758769 TI - Rationale for targeting interleukin-17 in the lungs. AB - An increasing body of evidence suggests that an exaggerated accumulation of activated neutrophils in the airways can cause decline of the clinical state in several lung diseases, including acute, severe asthma. In spite of this, an effective pharmacotherapy against this type of exaggerated neutrophil activity has yet to be developed. This review presents the scientific rationale for targeting the T-cell cytokine interleukin (IL)-17 in inflammatory lung diseases, to normalize neutrophil activity. Evidence leads to the conclusion that the production and release of IL-17 orchestrates neutrophil activity in the lungs. Hypothetically, local blockade of IL-17 may prove effective to normalize exaggerated neutrophil activity in lung diseases. PMID- 14758770 TI - Adenosine A2A receptor agonists as anti-inflammatory agents. AB - Stressed or injured tissues release endogenous adenosine, which blocks potentially destructive inflammatory cascades by binding to A2A adenosine receptors (ARs) and decreasing activation of platelets, leukocytes and endothelial cells. In these tissues, adenosine acts by reducing expression of adhesion molecules and release of pro-inflammatory mediators (e.g., reactive oxygen species, elastase and tumor necrosis factor-alpha). Synthetic A2A selective AR agonists are currently undergoing preclinical testing for the treatment of allergen-induced inflammation, ischemia-reperfusion injury, sepsis and autoimmune diseases. This review discusses potential disease targets for A2A AR agonist treatment, mechanisms of A2A AR agonist action and considerations for synthetic A2A AR agonist design. PMID- 14758771 TI - Individual cytokines contributing to asthma pathophysiology: valid targets for asthma therapy? AB - Asthma is a common, chronic inflammatory condition of the airways that leads to airway hyperresponsiveness, reversible narrowing of the airways, and airway wall remodeling. Cytokines are involved in various aspects of asthma pathophysiology, such as the polarization of T-helper (Th)2 cells, antigen presentation, immunoglobulin (Ig)E response, airway wall remodeling, and mast cell and eosinophil recruitment and activation. Th2-derived cytokines, such as interleukin (IL)-4, IL-5 and IL-13 contribute to many of these aspects. Inhibition of individual cytokines for asthma therapy has been, and continues to be investigated. Anti-IL-5 monoclonal antibodies did not demonstrate beneficial effects in asthma, with only partial inhibition of eosinophilia in the airway wall; soluble IL-4 receptor, which neutralizes the effects of IL-4, has provided modest improvements in moderate asthma. The anti-IgE monoclonal antibody approach has demonstrated the most benefit in allergic asthma, particularly in severe disease. Which individual cytokine target can be inhibited with beneficial effects comparable to or above that of current inhaled corticosteroids can only be discovered through clinical trials. Blocking the effects of more than one cytokine may be more successful, and greater therapeutic effects may be observed in particular categories of asthma. PMID- 14758772 TI - Leptin-based immune intervention: current status and future directions. AB - Current understanding of the role of leptin has expanded from its narrow association with obesity to a variety of effects on different biological processes including immune function. More specifically, leptin links nutritional status and energy balance to regulation of pro-inflammatory T-helper 1 immune responses. This has prompted several studies of targeted intervention with leptin antagonists in rodents to suppress onset and/or progression of chronic inflammation and autoimmunity. This review presents current preclinical evidence and potential applications for leptin-based immune approaches aimed at improving therapy for chronic and autoimmune conditions. PMID- 14758773 TI - The proteasome system and proteasome inhibitors in stroke: controlling the inflammatory response. AB - The ubiquitin-proteasome pathway (UPP) is a predominantly non-lysosomal protein degradation pathway responsible for degrading many critical regulatory proteins (e.g., nuclear factor-kappa B). This pathway is widely known for its ubiquitous role in immune and inflammatory responses, control of cell growth and apoptosis. These roles are apparent in the nervous system, but neurons and their neighboring cells also employ the UPP for distinct functions, ranging from development to the co-ordination of cellular responses, injury of the nervous system and brain specific processes such as aging and memory. Promising results from preclinical studies in animal models indicate that the use of specific proteasome inhibitors to manipulate UPP may prove valuable in treating such conditions as ischemic stroke. PMID- 14758774 TI - Prostanoid receptors and the urogenital tract. AB - Prostanoids are the metabolic product of cyclooxygenase-mediated metabolism of arachidonic acid. These prostanoids interact with a family of eight G protein coupled transmembrane receptors, which are abundantly expressed along the genitourinary tract and mediate the critical physiological actions of the prostanoids. These actions include modulation of renal glomerular hemodynamics, promotion of renal salt excretion, and maintenance of uroepithelial function and integrity. Normal functioning of prostanoid receptors is also a prerequisite for fertility and reproduction. Prostanoid receptor selective agonists and antagonists are likely to affect these processes quite differently from the simultaneous and global inhibition of all prostanoid synthesis that is achieved through the use of non-steroidal anti-inflammatory drugs. PMID- 14758775 TI - Natalizumab. Elan/Biogen. AB - Natalizumab is a humanized monoclonal antibody to alpha 4 beta 1 integrin (VLA-4) currently under development by Elan and Biogen for the treatment of Crohn's disease and multiple sclerosis. Phase II trials in both indications have been completed, and by December 2002 phase III trials in Crohn's disease and multiple sclerosis had been initiated. PMID- 14758776 TI - Semapimod. Cytokine. AB - Cytokine PharmaSciences is developing semapimod (CNI-1493), a cytokine inhibitor and synthetic guanylhydrazone mitogen-activated protein kinase blocker, as a potential treatment for Crohn's disease and other inflammatory conditions. As of December 2001, a phase I study demonstrating the safety of the compound had been completed and phase II trials for psoriasis and Crohn's disease were ongoing. In April 2003, preclinical and early clinical studies were underway for a variety of indications, including congestive heart failure and pancreatitis. PMID- 14758777 TI - Onercept. Serono. AB - Onercept is a recombinant human soluble p55 tumor necrosis factor binding protein under development by Serono for the potential treatment of a number of disorders, including Crohn's disease, psoriasis and psoriatic arthritis. In March 2002, phase II trials were ongoing for these indications, and in February 2003, Serono expected phase III trials in psoriatic arthritis to commence in the second half of 2003. PMID- 14758778 TI - Diquafosol tetrasodium. Inspire/Allergan/Santen. AB - Inspire, in collaboration with Allergan and Santen, is developing an eye-drop formulation of diquafosol tetrasodium (INS-365), a second-generation uridine nucleotide analog P2Y, receptor agonist for the potential treatment of dry eye disease. In June 2003, Inspire submitted an NDA for the treatment of dry eye, and in July 2003 the FDA granted the NDA Priority Review status. FDA action is expected in December 2003, and in January 2003 launch was expected in the first half of 2004. PMID- 14758779 TI - [Science of the nerves responsible for recognition in disorders of affective perception associated with schizophrenia]. PMID- 14758780 TI - [Pre-clinical study of disorders of information processing associated with schizophrenia]. PMID- 14758781 TI - [Schizophrenia and sex hormones]. PMID- 14758782 TI - [Psychotherapy of schizophrenia]. PMID- 14758783 TI - [Hypoglutamatergic hypothesis of schizophrenia: evidence from genetic studies]. AB - Schizophrenia is a relatively common but genetically complex disorder, making the identification of susceptibility genes formidable. However, progress in genetic studies on schizophrenia during the past ten years has revealed several replicated linkage loci, which span over multiple chromosomal regions. Since last year, several causal genes have been isolated from those linkage regions. All of these have proven to have some functional relevance to glutamatergic neurotransmission. These results are interesting because the "hypoglutamatergic hypothesis" for pathophysiology of schizophrenia has been articulated since the early eighties. This hypothesis has been supported both by pharmacological evidence that administration of NMDA-type glutamate receptor antagonists induces schizophrenia-like symptoms and by neurophysiological studies. Recent lines of evidence from a candidate gene approach have also endorsed the hypothesis. Here, we introduce the overview of recent progress in genetic studies that converge to depict the hypothesis of glutamatergic hypofunction in schizophrenia. PMID- 14758784 TI - [Efficacy of intervention with externalization therapy for eating disorders]. AB - Externalization has been one of the effective methods in the fields of brief therapy, family therapy, and psycho-education in recent years. In this study, we investigated the efficacy of intervention with externalization at the first stage of therapy in 25 patients with eating disorders. The subjects consisted of 11 patients with anorexia nervosa (AN) and 14 with bulimia nervosa (BN). The Eating Disorder Inventory (EDI) was evaluated at the first session, the 10th session, and six months later. The obtained results showed intervention with externalization resulted in significant decreases in not only total EDI score but also all the EDI subscale scores. We also found that there were great differences between the EDI subscale scores of anorexia nervosa and bulimia nervosa patients. Therapy was significantly less effective for patients with anorexia nervosa than for those with bulimia nervosa, and much less effective for the restricting type of anorexia nervosa. In addition, all the EDI subscale scores were significantly decreased, irrespective of the complication of personality disorder. The efficacy of intervention with externalization continued for six months. Especially in patients with anorexia nervosa, there were significant decreases in the EDI subscale scores when compared with the scores in the 10th session. The present findings indicates that initial intervention with externalization is effective for treating eating disorders, regardless of the severity of illness. PMID- 14758785 TI - [Antinociceptive mechanism of action of paracetamol]. AB - The mechanism of action of paracetamol (acetaminophen) is still not clearly understood. Unlike morphine, for example, paracetamol has no known endogenous high-affinity binding sites. In addition, paracetamol does not appear to share with nonsteroidal anti-inflammatory drugs (NSAIDs) the capacity to inhibit peripheral cyclo-oxygenase (COX) activity. There is currently considerable evidence to support the hypothesis of a central antinociceptive effect. Although various biochemical studies point to inhibition of central COX-2 activity, the existence of a COX activity that is selectively susceptible to paracetamol (COX 3?) is an alternative hypothesis. Modulation of the serotoninergic system has also been suggested on the basis of biochemical and behavioural studies supporting an indirect serotoninergic (5-HT) effect. Paracetamol may stimulate the activity of descending 5-HT pathways that inhibit nociceptive signal transmission in the spinal cord. Support for this possibility has come from evidence that spinally administered antagonists of several 5-HT receptor subtypes abolish the antinociceptive activity of paracetamol. These hypotheses have yet to be confirmed by further studies. Until then, the primary pharmacological mechanism underlying the analgesic effect of paracetamol has still to be clearly defined. PMID- 14758786 TI - [Pharmacologic basis for using paracetamol: pharmacokinetic and pharmacodynamic issues]. AB - The advantage of paracetamol (acetaminophen) is that it can be administered via the oral, intravenous or rectal routes. The last mentioned differs from the oral route in the slow and irregular absorption of the active substance. At therapeutic concentrations, the pharmacokinetics of paracetamol are linear--that is, independent of the dose, and constant with repeated administration. The efficacy of paracetamol has been demonstrated in a wide variety of acute or chronic painful syndromes. In adults, the optimum unit dose is 1 g. The maximum daily dosage is 4 g, consistent with the decline in analgesic activity, which is usually over 6 hours. With effervescent tablets, drug absorption and onset of action are more rapid than with conventional tablets. However, there is no direct correlation between serum concentrations of paracetamol and its analgesic or antipyretic effect. Paracetamol is the non-opiate analgesic of choice in elderly persons or patients with chronic renal insufficiency, and it is usually not necessary to reduce the dosage in such individuals, even though clearance is reduced. Although the bioavailability of paracetamol is not impaired in patients with chronic, benign liver diseases, the agent is contraindicated in those with hepatic insufficiency. It can be used during pregnancy and lactation. The very low level of paracetamol binding to plasma proteins, together with its hepatic metabolism, mainly through glucuronide or sulphate conjugation, account for the low risk of drug interactions with paracetamol, particularly with antivitamin K. When added to a traditional non-steroidal anti-inflammatory drug, paracetamol enhances the analgesic effect or allows the use of lower doses. It is more difficult to define the ideal dosage of paracetamol in children, because of the influence of age on its pharmacokinetics, and the relatively erratic bioavailability of suppositories. An oral dose of 15 mg/kg every 4 hours, up to a total of 60 mg/kg/day, is usually sufficient to achieve the desired analgesic or antipyretic effect. PMID- 14758787 TI - [Role of paracetamol in the acute pain management]. AB - Paracetamol (acetaminophen) has been shown to be an effective analgesic for the treatment of moderate pain where it is chiefly indicated, as shown in placebo controlled studies in the perioperative setting and other acute pain states. In addition, an opioid-sparing effect has been demonstrated. No clinically relevant adverse effects are usually apparent with recommended doses. Paracetamol is an effective component in 'multimodal analgesia' in combination with morphine, weak opioids and non-steroidal anti-inflammatory drugs. Although most studies involve the perioperative setting, similar results have been obtained in other acute pain states, such as acute musculoskeletal pain, migraine, etc. In conclusion, paracetamol has a favourable efficacy-tolerability profile and is therefore recommended as a basic, first-line analgesic in acute pain states and as a valuable component in multimodal analgesia. PMID- 14758788 TI - [Paracetamol in the treatment of osteoarthritis pain]. AB - Osteoarthritis (OA) is the most common joint disease and OA of the knee, in particular, is the major cause of chronic disability among people > 65 years. Because nonsteroidal anti-inflammatory drugs (NSAIDs) improve symptoms in many patients with OA, it is widely considered that OA pain is due to synovial inflammation. However, OA pain may arise also from subchondral bone, the joint capsule ligaments, tendons, entheses and periarticular muscle spasm. In many patients, the relief of OA pain and overall satisfaction with therapy may be as great with paracetamol (acetaminophen [APAP]) as with an NSAID. Cyclo-oxygenase (COX)-1-sparing NSAIDs (coxibs) are no more effective in the treatment of OA pain than non-selective NSAIDs and, although they may significantly decrease the risk of serious adverse effects related to gastrointestinal ulcers (GI) and ulcer complications, their gastroprotective effect may be reduced by concomitant administration of low-dose aspirin. Also, they may increase the risk of myocardial infarction in predisposed individuals. Because coxibs do not inhibit platelet aggregation, if prophylaxis against thromboembolic disease is required in patients being treated with a selective COX-2 inhibitor, low-dose aspirin should be used in conjunction with the coxib. Furthermore, nonselective NSAIDs and coxibs may have adverse effects on the kidney, fracture healing and salt and water homeostasis. This paper discusses the relative positioning of APAP, NSAIDs and coxibs in the management of OA, on the basis of considerations of tolerability, efficacy and costs. PMID- 14758789 TI - [Tolerability of paracetamol]. AB - Paracetamol (acetaminophen) is a well-tolerated drug at therapeutic doses and this safety profile is a major factor in the very wide use of the drug. It is well known that paracetamol is converted by the hepatic cytochrome P450 system to reactive compounds. Less well known is that paracetamol is also metabolized to the same reactive compounds by myeloperoxidase and the peroxidase function of cycloxygenase (COX)-1. The reactive metabolites lead to hepatotoxicity following overdosage. Similar hepatotoxicity has been reported after therapeutic doses, but critical analysis indicates that most patients with alleged toxicity from therapeutic doses have taken overdoses. Associations between the use of paracetamol and chronic renal diseases, gastrointestinal toxicity and asthma may be due to biases in case-control studies. In particular, biases may be caused by the perceived safety of paracetamol in these diseases. Selective inhibition of the delayed pathway of prostaglandin synthesis is consistent with the gastrointestinal safety of paracetamol and its safety in the majority of aspirin sensitive asthmatics. Despite the conversion of paracetamol to reactive compounds, hypersensitivity reactions are rare, although urticaria is produced in occasional patients. PMID- 14758790 TI - [Pain management: health economics and quality of life considerations]. AB - Pain represents a major clinical, social and economic problem, with estimates of its prevalence ranging from 8% to more than 60%, depending on the population. The impact of pain on economies is enormous, with the cost of back pain alone equivalent to more than one-fifth of one country's total health expenditure and 1.5% of its annual gross domestic product, while in another it represents three times the total cost of all types of cancer. However, decision makers have tended to concentrate their attention on one very minor component of the cost burden, namely prescription costs, which, in the case of back pain, represent 1% of the total cost burden. In addition to its economic impact, chronic pain is probably one of the diseases with the greatest negative impact on quality of life. For example, the quality of life for those with migraine had been shown to be at best equal to that for people with arthritis, asthma, diabetes mellitus or depression. The burden that pain imposes on individuals and the enormous costs that society has to bear as a result clearly demonstrate the need for collective thinking in the decision-making process. A broad, strategic perspective--based on evidence relating to effectiveness (including tolerability), efficiency and equity--is required in determining issues relating to the provision of services and resource allocation. In this regard, it is clear that paracetamol (acetaminophen) is effective in securing an analgesic effect; it has a good tolerability profile and is relatively cheap, in terms of both drug acquisition costs and its overall cost profile. Pain management strategies based on collective thinking should therefore place great reliance on paracetamol as an initial therapy in maximising pain relief and minimising cost and the impact of adverse effects. PMID- 14758791 TI - [New perspectives on paracetamol]. AB - Paracetamol (acetaminophen) is well established as a leading non-prescription antipyretic analgesic drug. Future developments are likely to include new formulations to achieve rapid absorption for a fast onset of action, and prolonged absorption to extend the duration of action for regular long-term administration. Better dosage forms are also required for rectal administration. The availability of intravenous paracetamol has greatly extended the use of this drug as an adjunct to postoperative analgesia and for control of fever in the intensive care setting. Intravenous paracetamol is available in only a few countries at present, but it seems inevitable that it will be marketed much more widely in the future. The misuse of paracetamol as a fashionable agent for self poisoning seems likely to continue, and liver failure may still occur in the small proportion of overdose patients who present too late for effective antidotal treatment with N-acetylcysteine. Much effort is being devoted to the study of the molecular mechanisms of paracetamol hepatotoxicity, and it is hoped that further advances may make it possible to prevent liver failure in all patients, irrespective of delays in presentation. At the same time, there is great interest in the mechanisms of the therapeutic actions of paracetamol and its effects on the different isoforms of cyclo-oxygenase. There will probably be important new findings in this area and these may lead to wider clinical use. Meantime, possible novel therapeutic applications for paracetamol include its use as an antioxidant to prevent atherosclerosis and cardiovascular disease by inhibiting the oxidation of low-density lipoproteins, and to prevent the formation of cataracts. PMID- 14758796 TI - New lights on green developments. PMID- 14758798 TI - State of the Journal. PMID- 14758797 TI - Effect of unstimulated saliva flow rate on experimental root caries. PMID- 14758800 TI - Endocytosis and pasta. PMID- 14758801 TI - The mitochondrial benzodiazepine receptor as a potential target protein for drug development: demonstration of functional significance with cell lines exhibiting differential expression of Bcl-2. AB - The article highlighted in this issue is "Reversal of Bcl-2 Mediated Resistance of the EW36 Human B-Cell Lymphoma Cell Line to Arsenite and Pesticide-Induced Apoptosis by PK11195, a Ligand of the Mitochondrial Benzodiazepine Receptor" by Donna E. Muscarella, Kerry A. O'Brien, Ann T. Lemley, and Stephen E Bloom from Cornell University in Ithaca, NY (pp. 66-73). The following brief review summarizes their findings, highlights the novel biological model and experimental approach used, and explores potential mechanistic and therapeutic implications of these findings. PMID- 14758802 TI - First trimester inhibin-A concentrations and later development of preeclampsia. PMID- 14758803 TI - Obstetric dilemma in an ovarian cancer patient. PMID- 14758804 TI - [Arthrosis: a common disease a little apart]. PMID- 14758805 TI - Visual vignette. Multiple endocrine neoplasia type 2A (MEN2A) associated with cutaneous lichen amyloidosis. PMID- 14758806 TI - Visual vignette. Riedel's fibrosing thyroiditis. PMID- 14758807 TI - Visual vignette. Bilateral adrenal calcifications. PMID- 14758808 TI - Medical image. Ruptured oesophagus. PMID- 14758810 TI - Whole-body MRI could replace bone scans in cancer patients. PMID- 14758809 TI - PET scans superior to CT for detection of colon cancer spread. PMID- 14758811 TI - [Idiopathic primary pulmonary hemosiderosis: treatment with cyclophosphamide and prednisone]. AB - The authors present the case of a child diagnosed as having idiopathic pulmonary hemosiderosis at five years of age who had a good clinical outcome at the age of ten years. Initially the patient was treated with prednisone and chloroquine with poor results. When cyclophosphamide was added to prednisone, the patient demonstrated clinical and radiological remission. To date, the patient has been followed for one year without any medication, and has had only one limited episode of hemosiderosis. The authors also suggest that the therapeutic regimen with cyclophosphamide and prednisone may be useful for some selected cases. PMID- 14758812 TI - [Special course for obtaining the nursing degree. Experience of the University of Padua]. AB - Padua's University introduced, in 2001/2002, a specific course of First Level Degree in Nursing, for conversion of precedent titles released by the National Health Service (Diploma in Nursing). Students had to attend 30 University Formative Credits (CFUs). 1298 students were enrolled in the curse. 293 students had no formative debt and have been directly admitted to final examination; 7 Those with a debt of 90 CFUs; 3 from 31 to 59 CFUs; 14 of 60 CFUs; 9 from 31 to 59 CFUs; 826 of 30 CFUs; 30 less than 30 CFUs; 99 of 26 CFUs; 17 less than 26 CFUs. Didactic activities were organized for student workers: on Fridays afternoon/evening and Saturdays. To facilitate didactic activities, these were transmitted during the night, by a private television of the region. Students received guidelines to product a final dissertation, and specific seminars. During the course students were given some questionnaires about satisfaction. 286 nurses (without formative debt) achieved the first level degree in the session of March 2002; 663 in the session of November 2002 and 56 in the session of March 2003, for a total of 1005 students. PMID- 14758814 TI - Modulation of PMN-endothelial cells interactions by cyclic nucleotides. AB - Polymorphonuclear leukocytes (PMN) interact with endothelial cells (EC) under normal and diseased conditions. Common Mechanisms exist regulating PMN-EC interactions in the systemic and pulmonary circulations and adhesion molecules play significant roles in both circulations, however there are important differences. Alterations in PMN deformability appear to be important in the pulmonary circulation because of the unique geometric and hydrodynamic conditions that exist in the pulmonary microvasculature. PMN work as the host's first line of defence against invading pathogens. Under certain circumstances, however, dysregulation of PMN-EC interactions may contribute to local or global tissue injury in diseases such as acute respiratory syndrome and multiple organ failure syndrome. Therefore, a thorough understanding of the regulation of PMN-EC interactions is important to understand the pathogenesis of this type of tissue injury, and modulation of PMN-EC interactions could be applicable to prevent or treat injury. cGMP and cAMP are cyclic nucleotides that work as second messengers and control numerous functions in PMN. This review covers the modulation of PMN EC interactions with cGMP and cAMP. Recent studies have shown that both cGMP and cAMP have inhibitory effects on events such as rolling, adhesion, migration and deformability change of PMN that are essential to PMN-EC interactions. Therefore, it is expected that the modulation of cyclic nucleotides is applicable for the treatment not only of local inflammatory diseases such as asthma but also of global tissue injury such as acute respiratory distress syndrome. PMID- 14758815 TI - The JAMIA Student Editorial Board: peer review education in biomedical informatics. PMID- 14758816 TI - On: Analysis of transference: a North American perspective. PMID- 14758817 TI - Uptake of BRCA1 genetic testing in adult sisters and daughters of known mutation carriers in Norway. AB - This study was undertaken to examine transmission of information to first-degree relatives of BRCA1 mutation carriers and uptake of genetic testing. The intention was to consider revision of current legislation related to privacy if information on life-saving health care was not disseminated to at-risk family members. The Norwegian Radium Hospital provides clinical genetics services for families at high risk for hereditary breast and ovarian cancer. Together with major hospitals nationwide we provide medical surveillance. Nearly all expenses are covered by the National Health insurance. Because of the high number of families with founder mutations in BRCA1, we are in a unique position to gather information about these groups. Within a consecutive series, we identified 75 BRCA1 mutation carriers and registered information transmission and uptake of genetic testing 6 months or more after the index mutation carriers had been informed about their mutation status. These 75 BRCA1 mutation carriers had 172 living first-degree relatives, aged 18 years or older (84 females, 88 males). Forty-four out of 54 (81.5%) of females over 30 had opted for genetic testing. The testing rate among all relatives was 43%. At any age, 63 % of the females underwent genetic testing compared with 24% of the males (p<0.05%). The overwhelming majority of adult females at risk opted for genetic testing. Males with daughters more frequently than males without daughters asked for testing. The findings give neither reason to reconsider legislation on privacy, nor for us to consider more aggressive methods of contacting relatives. PMID- 14758818 TI - Efficacy of low level laser therapy in reducing postoperative pain after endodontic surgery-- a randomized double blind clinical study. AB - The aim of the study was to evaluate the effect of low level laser application on postoperative pain after endodontic surgery in a double blind, randomized clinical study. Fifty-two healthy adults undergoing endodontic surgery were included into the study. Subsequently to suturing, 26 patients had the operation site treated with an 809 nm-GaAlAs-laser (oralaser voxx, Oralia GmbH, Konstanz, Germany) at a power output of 50 mW and an irradiation time of 150 s. Laser treatment was simulated in further 26 patients. Patients were instructed to evaluate their postoperative pain on 7 days after surgery by means of a visual analogue scale (VAS). The results revealed that the pain level in the laser group was lower than in the placebo group throughout the 7 day follow-up period. The differences, however, were significant only on the first postoperative day (Mann Whitney U-test, p<0.05). Low level laser therapy can be beneficial for the reduction of postoperative pain. Its clinical efficiency and applicability with regard to endodontic surgery, however require further investigation. This is in particular true for the optimal energy dosage and the number of laser treatments needed after surgery. PMID- 14758819 TI - Physician assistant as abortion provider: lessons from Vermont, New York, and Montana. PMID- 14758821 TI - Global AIDS treatment emergency. PMID- 14758820 TI - Analysis of 587 cases of oral squamous cell carcinoma in northern Thailand with a focus on young people. AB - The increasing incidence of oral squamous cell carcinoma (SCC) especially among younger people has been observed in many parts of the world. The objective of this study was to delineate the profile of patients with oral SCC with an emphasis on younger people in northern Thailand between 1991-2000. The medical records of 587 (median age 65, male-to-female ratio 1:3:1) patients presenting oral SCC were reviewed, with details of demographic data, staging, histological grading, treatment modality and risk factor profile being collated in detail. Seventy-five patients (12.8%) were 45 years of age or below (median age, 39 years). Most patients regardless of age had stage IV disease (56.2%). The most common histological gradings were well or moderately differentiated SCC (79.4%). The most common site regardless was tongue (42.8%). Most patients (79.4%) received treatments with either radiotherapy alone or a combination of surgery and radiotherapy. 64.4% of patients (87.2%) did not have a familial history of cancer. Collectively this data indicates that oral SCC remains a constant problem to the northern Thai population. In addition, the occurrence of oral SCC in young people is relatively high. Therefore, it is recommended that prevention of oral SCC with early detection, early treatment intervention, and withdrawal from risk habits are important factors for improving the wellbeing of these people. PMID- 14758822 TI - Technical considerations in distraction osteogenesis. AB - Five cases are presented to exemplify technical difficulties and complications which may be encountered when performing distraction osteogenesis in the facial skeleton. The procedure should be performed under close supervision by the surgical and orthodontic colleagues. Errors in the choice of vector may be managed by earlier removal of the distractor and subsequent traction on the previously osteotomized segments using orthodontic appliances and principles. Multiple distractors may be inserted in the same jaw and bimaxillary procedures are possible, increasing the likelihood of encountering technical difficulties. Detailed planning and close follow-up, with early recognition and active management of the complications, may be useful in ensuring a successful outcome of this versatile procedure. PMID- 14758823 TI - Unsung hero. PMID- 14758824 TI - The mouth's defense against HIV discovered. PMID- 14758825 TI - USA strengthens its BSE controls. PMID- 14758826 TI - Residues surveillance results. PMID- 14758827 TI - Footrot and lameness in sheep. PMID- 14758828 TI - Prevalence of benzimidazole resistance on horse farms in Germany. AB - Faecal egg counts (FECs) were made on samples from 1383 horses on 64 farms in northern Germany between August 2000 and November 2001. There were significant differences between the mean FECs in the two years; in 2000, 59.6 per cent of 369 samples were positive and in 2001, 32.6 per cent of 1014 samples were positive for strongyle eggs. The results of a FEC reduction test indicated that resistance to fenbendazole was present on all 10 farms where it had been used, including in 33 of 60 horses tested. In contrast, treatment with ivermectin resulted in the complete elimination of nematode eggs in all the 77 horses tested. The mean LD50 values of the egg hatch test for thiabendazole indicated resistance on all 20 farms investigated and in 94 of 134 samples (70 per cent). PMID- 14758829 TI - Development of the skeleton and feathers of dusky parrots (Pionus fuscus) in relation to their behaviour. AB - A clutch of five dusky parrots (Pionus fuscus) was observed from hatching to fully grown. They were examined radiographically from 16 to 45 days of age, a few days before the cessation of bone growth, and the development of their feathers and their behaviour were also studied. It was observed that when growing birds were removed from the nest and placed singly on a flat surface they would stand up and walk about until restrained; normally these birds would move very little and lie in an intertwined huddle that supported their relatively weak growing skeletons. At 50 days old they would climb to the nest entrance, retreating if scared. From day 51 the parrots flapped their wings vigorously inside the nest box, and they emerged at 53 days old when nearly all their large feathers had finished growing. These findings may help to explain the high rate of juvenile osteodystrophy in hand-reared parrots; premature exercise could lead to pathological deformity of the long bones, especially the major weight-bearing bone, the tibiotarsus. PMID- 14758830 TI - Periparturient increase in faecal egg counts in a captive population of mohor gazelle (Gazella dama mhorr). AB - The objective of this study was to assess whether there was a periparturient rise in the faecal egg output of a population of North African gazelles (Gazella dama mhorr) kept in captivity in Almeria, southern Spain. In one experiment faeces were collected from 47 female gazelles on three days in winter, in November and December 1995 and January 1996; in a second experiment faecal samples were collected from nine pregnant gazelles at weekly intervals from July 1996 to June 1997. The mean trichostrongylid faecal egg counts were significantly higher (P < 0.05) in the periparturient gazelles than in the pregnant and non-pregnant animals only when the births took place in winter. Other factors, including the gazelle's age, its level of inbreeding, the number of previous births, and its trichostrongylid egg output at the beginning of the study did not affect whether it showed a periparturient rise. The parasites responsible for the rise were different in the two experiments. PMID- 14758831 TI - Different outcome of intrauterine infection with bovine viral diarrhoea (BVD) virus in twin calves. PMID- 14758832 TI - Signs of spinal cord disease in two heifers caused by Listeria monocytogenes. PMID- 14758833 TI - Biochemical characterisation of Francisella tularensis strains isolated in Spain. PMID- 14758834 TI - Prevalence of Salmonella shedding in faeces by captive chelonians. PMID- 14758836 TI - Prescribing and dispensing of veterinary medicines. PMID- 14758835 TI - Thiamine deficiency in sheep with chronic rumen acidosis. PMID- 14758837 TI - BARBs and the genetics of BSE. PMID- 14758838 TI - Continuing anxiety about BSE. PMID- 14758839 TI - Practice standards. PMID- 14758840 TI - Anthelmintic resistance and use of anthelmintics in horses. PMID- 14758842 TI - Identification and microchips. PMID- 14758841 TI - Anthelmintic resistance and use of anthelmintics in horses. PMID- 14758843 TI - Identification and microchips. PMID- 14758844 TI - Survey of purebred dogs in the UK. PMID- 14758845 TI - The development of molecular compartmentation in the cerebellar cortex. AB - The cerebellum is subdivided into hundreds of discrete modules defined by their connectivity and molecular signatures. Cerebellar compartmentation arises very early in development through the formation of multiple populations of chemically distinct Purkinje cells that migrate in a coordinated fashion to form parasagittal bands of cells. Different Purkinje cell bands are then innervated by discrete subpopulations of cerebellar afferents. Because of its stereotyped and strikingly beautiful organization the cerebellum is an excellent model in which to explore genetic/epigenetic aspects of pattern formation in the central nervous system. PMID- 14758846 TI - Differentiation of oxyntic cells during early ontogenesis in the bovine. AB - The origin and the differentiation of oxyntic cells in fetal bovine abomasum were investigated using transmission electron and light microscopic methods. In the oxyntic gland region oxyntic cell precursors and immature oxyntic cells appear as early as at the end of the first trimester of gestation--much earlier than described in any other mammalian animal species. Immature oxyntic cells are characterized by long apical microvilli, by their triangular-shaped light cytoplasm rich in large and numerous mitochondria, by the existence of vesicular profiles and by the incipient invagination of the apical plasma membrane forming a primitive intracellular canaliculus expanding into central areas of the cell. The oxyntic cell represents the first exocrine cell type developing from secretory granule-containing cells in the base of the primitive gastric glands. PMID- 14758847 TI - Direct vs. indirect effects of trypan blue on the ectodermal cells of the 11.5 day rat embryo? AB - The effect of trypan blue on the 11.5-day rat conceptus after intravitelline vessel administration is described. For comparison, conceptuses injected with varying volumes of Hanks' BSS have also been studied. Trypan blue significantly retarded the growth and development of conceptuses after 6 hours incubation in vitro. The SEM revealed rounded ectodermal cells, some of which appeared disrupted. These cells seemed to cause some of the intersomitic grooves to disappear, making a number of the somites indistinct from the outside. Unlike cells of uninjected embryos, the surfaces of the affected ectodermal cells lacked microvilli and their perimeters were lined with microvilli-like structures which appeared matted together. It was concluded that trypan blue affected the embryo directly probably by disturbing its fluid and ionic balance. PMID- 14758848 TI - Lectin-induced deciduoma formation in the pseudopregnant rat. AB - Decidual cell induction in the pseudopregnant rat was examined in this study using the lectin concanavalin A (ConA). The histochemical binding of the lectin to the uterine cell surface at the time of deciduomatic induction was also studied. ConA was found to induce significant deciduomata (decidual-like tissue) in the uterine horn when injected intraluminally on day 5 of pseudopregnancy (PSP). ConA-induced deciduomata appeared as a series of discrete nodules in the uterine horn, reminiscent of the anatomical appearance of normal embryo implantation sites. Deciduoma induction by ConA was greatly reduced by pre absorption of the lectin with its competitive sugar. Lectin histochemistry revealed binding of ConA to the cell surface on day 5 of PSP. Pre-absorption of the lectin with its competitive sugar also significantly reduced surface binding of the lectin, and this finding may be correlated with the greatly reduced ability of the pre-absorbed lectin to induce deciduomata. Possible mechanisms for the induction of deciduomata by lectins are considered. PMID- 14758849 TI - Relationship between neuropeptide Y and luteinizing-hormone-releasing hormone immunoreactivities in the hypothalamus and preoptic region. AB - The purpose of this study was to examine the anatomical relationships of perikarya and fibers containing neuropeptide Y (NPY) and luteinizing-hormone releasing hormone (LHRH) in the hypothalamus and preoptic region of female rats. In view of our previous report of stimulatory effects of estrogen on LHRH and NPY levels in the median eminence, animals were bilaterally ovariectomized and subsequently implanted subcutaneously with capsules containing estradiol benzoate in oil or vehicle. Following intracerebroventricular injection of colchicine, rats were perfused with fixative and their brains sectioned and processed for immunohistochemical visualization of NPY and LHRH in the same section and in consecutive sections. Estrogen treatment had no discernible effect on the distribution or relationship of these peptides. NPY-immunoreactive fibers were intimately associated with LHRH-labeled primary dendrites and perikarya in the medial preoptic region and horizontal limb of the diagonal band of Broca. Fibers containing NPY or LHRH overlapped extensively in the lateral palisade region of the median eminence and also in the subependymal and internal zones. The external zone of the median eminence displayed relatively less overlap of these peptide systems. LHRH-immunoreactive axons coursed among NPY-labeled perikarya in the arcuate nucleus and appeared to contact these cells. These results suggest that NPY-containing axons may influence LHRH-positive neurons at the cell body and also at the site of axon termination in the median eminence. LHRH-containing axons appear to contact NPY-immunoreactive perikarya in the arcuate nucleus and may interact with terminals in the median eminence. This arrangement may provide a mechanism for communication between NPY and LHRH neurons and for the neuroendocrine coordination of hypothalamic NPY and LHRH secretion before ovulation. PMID- 14758850 TI - Dendritic arbor alterations in the medial superior olivary neurons of neonatally underfed rats. AB - Golgi-Cox-stained bipolar cells of the medial superior olive (MSO) were analyzed in control and undernourished Wistar strain rats at 12, 20, 30 and 40 days of age. Undernutrition significantly reduced the number of dendrites and the extension of ipsilateral dendritic prolongations, with no effects upon the cross sectional somal area and minimal alterations in the corresponding contralateral dendritic branches. The data suggest that in underfed rate, afferents from the receptors projecting to the MSO via the anteroventral cochlear nuclei may cause an imblance in the binaural interactions which occur between the axon terminals and the ipsilateral contralateral dendritic arbors of MSO neurons. PMID- 14758851 TI - Fine structural distinction between ganglia of the outer and inner submucosal plexus in porcine small intestine. AB - Ultrastructural differences between ganglia of the plexus submucosus internus (Meissner; PSI) and plexus submucosus externus (Schabadasch; PSE) are described. Comparison revealed a different glia index (ratio glia per neuron) between the PSE (3:1) and the PSI (1:1), the arrangement of PSI neurons in compartments and the appearance of broad membrane-to-membrane appositions inside the compartments of the PSI. Structural and immunohistochemical differences between the two plexuses are discussed. In general, PSE neurons show a wider variety in size and shape than most of the PSI neurons. PMID- 14758852 TI - A morphological study of the primary cilia in the rat pancreatic ductal system: ultrastructural features and variability. AB - Primary cilia in the pancreas of the rat were studied by transmission electron microscopy. Their presence is very common, and each ductal cell seems to be provided with a single cilium. The basal body showed anchoring apparatus such as transitional fibers and basal feet. The shaft can show a number of different patterns according to the level of the sections. Proceeding towards the tip, the microtubules decrease in number, although not always in the same way. Near the tip, it is possible to detect patterns, with only 1 microtubule. Three kinds of tips are described. The function of the cilia is discussed. PMID- 14758853 TI - Arteriographic study of the arterial supply of the foot in one hundred cadaver feet. AB - The arterial supply of 100 human cadaver feet (87 cadavers) was investigated by stereoscopic arteriography and was compared phylogenetically to that of the macaque foot. The deep plantar arch was always well developed and complete, whereas the superficial plantar arch was usually slender and incomplete. The first proximal perforating artery arising from the dorsalis pedis artery formed the main component of the deep plantar arch in 82% of the feet. The second proximal perforating artery arising from the dorsal rete contributed to the deep plantar arch in 43% of the feet, and formed most of the arch in one foot. The dorsal rete was classified into four groups of variants based on the arterial source of the second dorsal metatarsal artery. These were the arcuate artery (25%), distal lateral tarsal artery (12%), proximal lateral tarsal artery (6%), and nondorsal rete (57%) variants. In the first intermetatarsal space, the dorsal and plantar metatarsal arteries shared a common trunk in 54% of the feet, but this did not occur in the other intermetatarsal spaces. The second dorsal metatarsal artery arose from the dorsal rete in 43% of the feet, and this artery was quite large, sometimes being the largest of all the dorsal and plantar metatarsal arteries. Variations of the arterial supply found in humans sometimes resembled the typical pattern found in the macaque. PMID- 14758854 TI - Individual and contextual determinants of inequalities in health: the Italian case. AB - The geographic distribution of health status across Italian regions shows a North South gradient, with better conditions in the North for both males and females. Using data from the 2000 National Health Interview Survey, the authors first analyze the geographic variation in subjective health and presence of chronic conditions, with specific attention to the effects of individual and area-based socioeconomic conditions and their heterogeneity across regions. The results suggest the North-South gradient in health is mainly affected, at least for subjective health, by the different composition of macro-areas with respect to individual education, and is slightly influenced by contextual circumstances. Moreover, being less educated results in poorer health in some regions (mainly South and Isles) than in others (mainly Northeast). The authors next analyze the circumstances affecting the presence of more disadvantaged people in the South, to highlight features of the Southern context that might exacerbate social inequalities in health and features of Northern areas that might allay them. Indicators of inequalities, welfare, labor, and power resources were analyzed. The results confirm the disadvantage of the South in terms of social, economic, and cultural features, mainly revealing the compositional effects found in the first part of the study. However, the contextual predictive value of income inequalities, quality of care, and social cohesion can have a supplementary effect on health outcomes of disadvantaged persons. PMID- 14758855 TI - Regional differences in trends in life expectancy and the influence of the political and socioeconomic contexts in Germany. AB - The aim of the study is to investigate to what extent trends in life expectancy are influenced by political variables and socioeconomic characteristics that play a role at the regional level of the federal states in Germany. Data on life expectancy in males and females at birth are analyzed from 1986 to 1998 for 12 federal states in Eastern and Western Germany. These states are classified into five types of political government since 1980: (1) long-term Christian democratic, (2) long-term social democratic, (3) change from Christian to social democratic, (4) change from communist to social democratic, and (5) change from communist to Christian democratic. The study showed three main results. First, life expectancy has been directly influenced by the major political forces that determined policies in East and West Germany. Second, life expectancy was higher in federal states with predominantly Christian democratic governments than in those with predominantly social democratic governments. Third, life expectancy was strongly related to the economic power of the federal states. Because federal states characterized by a more prosperous economic situation were those with a predominantly Christian democratic government, while federal states with a less prosperous situation were mostly governed by social democrats, it is difficult to disentangle the effects of economic and political factors on life expectancy. Nevertheless, this study underlines the importance of politics and policies on such robust and more general health indicators as mean life expectancy at birth. PMID- 14758856 TI - Power relations and premature mortality in Spain's autonomous communities. AB - This trends ecological study analyzes, across 17 autonomous communities of Spain from 1989 to 1998, the relationship between mortality (total and by main causes of death) and power relations (type of government: social democratic (SDP), conservative (CDP), and others), labor market variables, welfare state variables, income inequality, absolute income, poverty, and number of civil associations. The authors conducted a descriptive analysis; a bivariate analysis (Pearson correlation coefficients) between mortality and each of the independent variables; and a multivariate analysis, adjusting multilevel linear regression models. All dimensions of the conceptual power relations model were related to premature mortality in the direction hypothesized. The cross-pooled multilevel regression models show that total premature mortality in males, male and female cerebrovascular mortality, male and female cirrhosis mortality, and male lung cancer mortality decreased somewhat more in communities where primary health care reform was implemented more quickly. Premature mortality decreased somewhat more in SDP than in CDP communities for male and female total premature mortality, cerebrovascular mortality, and cirrhosis mortality, and male lung cancer mortality. These results are in accord with earlier studies that found a relationship among health indicators and variables related to labor market, welfare state, income inequalities, civil associations, and power relations. PMID- 14758857 TI - Social differentials in the decline of infant mortality in Sweden in the twentieth century: the impact of politics and policy. AB - This article describes some of the policies behind the decline of infant mortality in Sweden during the 20th century, from very high levels and large social differentials at the turn of the 19th century to one of the lowest levels in the world by 1950. Political commitment to reducing infant mortality and disparities between groups, a more equitable distribution of economic resources, and a successful combination of universal social and health policies most benefiting the least advantaged families and their children contributed to this favorable development. PMID- 14758859 TI - The beryllium "double standard" standard. AB - Brush Wellman, the world's leading producer and supplier of beryllium products, has systematically hidden cases of beryllium disease that occurred below the threshold limit value (TLV) and lied about the efficacy of the TLV in published papers, lectures, reports to government agencies, and instructional materials prepared for customers and workers. Hypocritically, Brush Wellman instituted a zero exposure standard for corporate executives while workers and customers were told the 2 microgram standard was "safe." Brush intentionally used its workers as "canaries for the plant," and referred to them as such. Internal documents and corporate depositions indicate that these actions were intentional and that the motive was money. Despite knowledge of the inadequacy of the TLV, Brush has successfully used it as a defense against lawsuits brought by injured workers and as a sales device to provide reassurance to customers. Brush's policy has reaped an untold number of victims and resulted in mass distribution of beryllium in consumer products. Such corporate malfeasance is perpetuated by the current market system, which is controlled by an organized oligopoly that creates an incentive for the neglect of worker health and safety in favor of externalizing costs to victimized workers, their families, and society at large. PMID- 14758860 TI - Reflections on the twenty-fifth anniversary of the Alma-Ata Declaration. AB - The Alma-Ata Declaration on Primary Health Care of 1978-based on the World Health Assembly's resolution of 1977 on Health for All by the Year 2000--was a watershed in the concepts and practices of public health as a scientific discipline; it was endorsed by every country in the world, rich and poor. According to the Declaration, health is a fundamental right, to be guaranteed by the state; people should be the prime movers in shaping their health services, using and enlarging upon the capacities developed in their societies; health services should operate as an integral whole, with promotive, preventive, curative, and rehabilitative components; and any western medical technology used in non-western societies must conform to the cultural, social, economic, and epidemiological conditions of the individual countries. Since Alma-Ata, a syndicate of the rich countries and the ruling elites of the poor countries, aided by the WHO, World Bank, World Trade Organization, and other international institutions, has done much to overturn the Declaration's primary health care initiatives. The WHO's recent attempt to regain some credibility, its Commission on Macroeconomics and Health, ignored the primary health care principles of the Alma-Ata Declaration. A struggle for these principles will have to be part of the larger struggle, by like-minded individuals working in individual countries, for a just world order. PMID- 14758858 TI - Conflicts between commercial and scientific interests in pharmaceutical advertising for medical journals. AB - In 1992, researchers from the University of California, Los Angeles, published a study on the scientific merit and validity of pharmaceutical advertisements in medical journals. Their results led them to conclude, provocatively, that many pharmaceutical advertisements contained deficiencies in areas in which the U.S. Food and Drug Administration had established explicit standards of quality. This article provides a detailed account of third-party reactions to the study following its publication in the Annals of Internal Medicine, as well as the implications for those involved, including the authors, editors, and publisher. The increasingly diverging interests between medical journal editors and publishers are also discussed and highlighted by two recent cases of editors' departures from prominent general-interest medical journals. PMID- 14758862 TI - Political context of the World Health Organization: sugar industry threatens to scupper the WHO. AB - The Sugar Association, representing the U.S. sugar industry, is highly critical of a WHO report on guidelines for healthy eating, which suggests that sugar should account for no more than 10 percent of a healthy diet. The association has demanded that Congress end its funding of the World Health Organization unless the WHO withdraws the guidelines, and the association and six other big food industry groups have also asked the U.S. Secretary of Health and Human Services to use his influence to get the WHO report withdrawn. The WHO strongly rejects the sugar lobby's criticisms. PMID- 14758861 TI - The politics of underdevelopment: metered to death-how a water experiment caused riots and a cholera epidemic. AB - Water privatization programs in South Africa, part of a government policy aimed at making people pay for the full cost of running water ("total cost recovery"), was developed by private water companies and the World Bank to finance improved water supplies and build the country's economy. Instead the programs are causing more misery than development. Millions of poor people have had their water supply cut off because of inability to pay, forcing them to get their water from polluted rivers and lakes and leading to South Africa's worst cholera outbreak- which the government paid millions of dollars to control. Residents in some townships are rebelling, and many of the private multinational water companies are reassessing their involvement in South Africa. PMID- 14758863 TI - A review of data on the U.S. health sector. June 2003. AB - This report presents information on the state of the U.S. health sector in early 2003. It provides data on the insured and uninsured and their access to health care; the increasing costs of care; the role of corporations--including the hospital, pharmaceutical, and insurance industries--and corporate money in health care; and Medicare HMOs. The author provides updates on health care policy and proposals at the state (including single-payer bills) and federal levels; the Bush proposals on Medicare and Medicaid; and the health care reform proposals put forward by the Democratic presidential candidates. PMID- 14758864 TI - Acrylamide in heat-processed foods--a carcinogen looking for human cancer? PMID- 14758865 TI - Prevention comes of age? PMID- 14758866 TI - Berkson's bias reviewed. PMID- 14758867 TI - Studying risk factors for Parkinson's disease. PMID- 14758868 TI - The epidemiology of lifestyle and risk for type 2 diabetes. AB - Results from ecological and migration studies indicate that a western lifestyle is associated with a higher prevalence of type 2 diabetes. In recent years, there has been a rapid accumulation of data on lifestyle and risk for type 2 diabetes from studies on an individual level. This article gives an overview of the evidence for the effect of different lifestyle factors on risk for type 2 diabetes and discusses various methodological approaches. Randomized intervention studies have shown that changes in diet and physical activity can protect against type 2 diabetes. Diet and physical activity can affect the development of type 2 diabetes through changes of body fatness, but also through other pathways. Findings from cohort studies and trials with intermediary endpoints have indicated that higher consumption of whole grain products and exchanging unsaturated fat for saturated fat may lower risk for type 2 diabetes. In addition, several new promising hypotheses about diet and the development of type 2 diabetes are currently being investigated. Light to moderate alcohol consumption may also reduce risk for type 2 diabetes, whereas high alcohol consumption and cigarette smoking may increase risk for type 2 diabetes. Prevention of weight gain by balancing energy intake and expenditure is of paramount importance to limit current increases in the prevalence of type 2 diabetes. In addition, other effects of lifestyle may play an important role in reducing risk for type 2 diabetes. PMID- 14758870 TI - Case-control study of environmental risk factors for Parkinson's disease in Belgium. AB - The aetiology of Parkinson's disease (PD) is unknown and said to be multifactorial. We report on a retrospective epidemiological case control study, performed in Flanders during a 3-year period, investigating known and potential environmental risk factors for PD by means of questionnaires. We investigated 423 prevalent patients and 205 spouse-controls. We found familial occurrence in 15% of the patients, a mean age of onset of 58 years, and a clear male preponderance (male/female ratio 1.53). Our results suggest more nulliparity among female PD patients (95% CI: 1.08-5.76). We found a discrete clustering of patients in areas with intensive metallurgic frequently employed in metallurgy than controls (95% CI: 1.04-9.20). Furthermore, patients were clearly more exposed to zinc (95% CI: 1.51-90.90) and toluene (95% CI: 1.03 58.82). Male patients report more prostatectomy-surgery (95% CI: 1.54-17.24). PMID- 14758869 TI - The limitations of using hospital controls in cancer etiology--one more example for Berkson's bias. AB - The aim of this report was to present an example in which Berkson's bias, most probably, affected the results of a study by overriding the influence of a well established risk factor (smoking) in the etiology of bladder cancer. The results of a study of 140 male patients with bladder cancer and 280 matched hospital controls confirmed the etiological role of industrial occupation in bladder cancer but failed to confirm the role of smoking. We reanalyzed the proportion of chronic related morbidity as well as the rate of smoking in patients with lung disease in cases and controls. A similar distribution of some chronic diseases known to be highly associated with smoking was found among cases and controls. Highest smoking rates (91%) were found among patients with bladder cancer who also reported a concomitant lung disease, and the lowest rate (67%) was noted among controls without lung disease (p = 0.009). Using the prevalence of smoking in the general Israeli male population (50%), significant odds ratio for bladder cancer among ever smokers compared to never smokers was observed. Our conclusion is that a possibility of Berkson's bias should be considered whenever hospital controls are used. Information on diseases related to the risk factor under consideration and on the prevalence of the risk factor in the general population, may demonstrate the existence of such a bias. PMID- 14758871 TI - The epidemiology of diabetes in a large Israeli HMO. AB - Diabetes is one of the most prevalent non-communicable disease globally and it is one of the leading cause for death in most developed countries. The current population-based study aim was to describe to the epidemiology of diabetes in Israel by using our HMO's automated medical databases. All diabetic patients appearing in the diabetes registry among 1.6 million insured members in the second largest HMO in Israel were selected for epidemiological analysis. We identified 39,768 diabetic patients (crude prevalence rate of 2.6%). Higher age specific prevalence rates were recorded among males. The highest age-specific prevalence rate of diabetes was calculated for men aged 75 and above (18.1%). A rise in the prevalence and mortality rates was recorded between 1999 and 2001 female (from 1.9 to 2.8 per 100,000) and for men (from 2.3 to 3.8 per 100,000). The current study demonstrates the potential of using large automated medical and administrative databases to determine the epidemiology of chronic disease, such as diabetes. The rise in the prevalence and mortality of diabetes patients has important implication for Israeli health authorities and should be seriously regarded. PMID- 14758872 TI - Serological survey of immunity to tetanus in adult population of Northern Halkidiki, Greece. AB - BACKGROUND: Despite the implementation of the obligatory anti-tetanus vaccination, the tetanus cases in Greece are not eliminated. Because of the increased possibility of Clostridium tetani infection of the Northern Halkidiki population--like any other rural population the evaluation of the immunity to tetanus in the area is considered necessary. METHODS: The tetanus antitoxin levels were determined using the enzyme-linked immunosorbent assay (ELISA) in 405 healthy adult individuals attending the health center for routine laboratory tests. RESULTS: 64.4% of the studied population was found protected (tetanus antitoxin levels > or = 0.1 IU/ml). The percentage of protected people decreased as age increased from 83.3% in the 21-30 to 51.2% in the > 60 age group. 82.1% of the tested males and 52.6% of the tested females had tetanus antitoxin levels > or = 0.1 IU/ml (p < 0.0001). The percentages of immune men (100-66.2% in various age groups) were found higher than those of women (80.8-35.5% in the respective age groups). The geometrical mean titres (GMTs) were 0.44 in all of 261 immune people, 0.53 in 133 immune men and 0.37 in 128 immune women (p = 0.0021). CONCLUSION: The proportion of protection among men over 60 and women over 30 years old is inadequate, the levels of tetanus antitoxin decline with age and a significant difference was found between the proportion of protection of males and females. PMID- 14758873 TI - Considering the antimicrobial sensitivity of the intestinal botulism agent Clostridium butyricum when treating concomitant infections. AB - In Italy, neurotoxigenic Clostridium butyricum has been reported as a new agent of intestinal toxemia botulism, and most of the cases have been associated with enterocolitis. Although infections concomitant with botulism must be treated with antibiotics, this can increase the severity of botulism. We discuss the sensitivity of this agent to certain antibiotics, compared to findings on the sensitivity of C. botulinum. PMID- 14758874 TI - Characterization of Borrelia burgdorferi sensu lato from Novosibirsk region (West Siberia, Russia) based on direct PCR. AB - Borrelia burgdorferi sensu lato infecting Ixodes persulcatus ticks near Novosibirsk, Russia were detected using PCR with primers specific to 5S and 23S rRNA genes. Two genospecies, B. afzelii and B. garinii, were identified by the PCR-based restriction fragment length polymorphism analysis with Tru9-I restriction endonuclease. Comparison of the corresponding nucleotide sequences revealed considerable diversity of the 5S-23S intergenic spacer structure among B. garinii. PMID- 14758875 TI - An outbreak of Yersinia enterocolitica O:3 infections on an oil tanker. AB - In January 2002, an outbreak of Yersinia enterocolitica O:3 infections occured on 'Asirat', an oil tanker, during its return test voyage from Split, Croatia to Trieste, Italy. Of the 120 crewmembers and workers 22 (18%) suffered from gastrointestinal symptoms. In 17 patients Y. enterocolitica O:3 was isolated from stool samples. All available food and water samples were negative and the source of infection was not determined. Probably a foodborne transmission was involved, although person to person transmission could not be excluded. PMID- 14758876 TI - Changes in the epidemiological pattern of Mycoplasma pneumoniae infections in Poland. PMID- 14758877 TI - Trends of weight, height and obesity in young Portuguese males: 1995-1999. PMID- 14758878 TI - Circulating intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1) and E-selectin in urological malignancies. AB - Several studies suggest that cellular adhesion molecules (CAM) play a role in cancer progression and metastasis. To evaluate the role of these molecules as possible tumor markers in patients with urological malignancies, we examined the serum levels of intercellular cell adhesion molecule-1 (ICAM-1), vascular cellcular adhesion molecule-1 (VCAM-1) and E-selectin in patients with renal cell , bladder-, prostate- and testicular cancer. Serum levels of 237 patients with urological cancers, renal cell carcinoma (n = 47), bladder cancer (n = 81), prostate cancer (n = 87) and testicular cancer (n = 22) and a group of 41 patients with benign prostate hyperplasia (BPH) as well as a 42 healthy control persons were examined for CAMs by specific ELISA tests. Serum CAM concentrations of all tumor patients were compared with controls and within the group according to T stage, N stage, tumor grade and extent of distant metastasis. Our results demonstrate that ICAM-1 and VCAM-1 serum levels are not stage dependently elevated; in contrary, they demonstrate a wide range and are highly variable throughout the different cancer types. In renal cell cancer and in bladder cancer, there is a significant difference for ICAM-1 between controls and T3 and T4 and metastatic cancers. A similar difference was found for VCAM-1, however not for E-selectin in any tumor group. Testicular cancer and prostate cancer did not demonstrate any difference in CAM serum levels between patients with tumors and controls. In metastatic renal cell-, bladder- and prostate cancer, the serum levels of ICAM-1 and VCAM-1 showed a tendency to correlate with the extent of metastatis although no statistical difference between patients with a single metastatic lesion and patients with multiple lesions could be demonstrated. The results of this study implicate a rather limited role of cellular adhesion molecules. Despite of significant ICAM-1 or VCAM-1 serum levels in some locally advanced tumors or metastatic disease, this observation does not provide enough relevant clinical information for use as tumor markers. PMID- 14758879 TI - Is incidence of cancer on the decline in Delhi, capital of India? AB - The incidence of cancer varies to a great extent in the resident population of Delhi (Urban area). The purpose of this study is to examine the time trends of incidence rates in different sites (ICD.9) of cancer. Average Annual Age-Adjusted (world population) incidence rates by site (ICD.9) is used to estimate the parameters by linear regression method. Time trends analysis of 8 years from 1988 to 1995 in Delhi for age-adjusted, truncated (35-64 years) incidence rates to world population and crude incidence rate by site (ICD.9) did not reveal statistically significant decrease or increase in the incidence. PMID- 14758880 TI - Port site metastasis subsequent to laparoscopy for chronic pelvic pain. AB - We present an interesting case of port site metastasis is a menopausal women subsequent to diagnostic laparoscopy undertaken for chronic pelvic pain, which later proved early ovarian malignancy as the source of primary. While cases of port site metastasis have mostly occurred after extensive disease the possbility of such complication should be in mind at laparoscopy of early cancer. PMID- 14758881 TI - Fusarium solani fungemia in a patient with acute lymphoblastic leukemia. AB - A patient with ALL on anticancer chemotherapy developed fever which was later attributed to be due to Fusarium fungemia. The details of the case & a review of literature follows. PMID- 14758882 TI - Thalidomide: from teratogen to anti-angiogenic. AB - Thalidomide, infamous for its teratogenic potential is now emerging as a therapeutic option for many disorders owing to its immunomodulant and anti angiogenic effect. The demonstration of its ability to inhibit experimental angiogenesis in corneal micro-pocket assay in experimental animals led to the speculation that it might prove to be effective in certain angiogenesis dependent tumors. However, contrary to expectations, it failed to prevent tumor growth in certain animal models. The clinical study evaluating the effects of thalidomide in large number of patients suffering from refractory myeloma have shown encouraging results. Further trials are in progress to evaluate its effects in various other malignancies. In this review, various experimental and clinical studies demonstrating anti-angiogenic and anti-tumor effect of thalidomide are discussed with emphasis on its mechanism of action. PMID- 14758883 TI - Vertical rectus abdominis myocutaneous flap cover for lower abdomen, chest wall, groin and thigh defects following resection of malignant tumours. AB - Vertical rectus abdominis mycutaneous (VRAM) flap provides a reliable flap cover for large soft tissue defects of chest wall, torso, groin, perineum and thigh. It has been mainly used in trauma and benign conditions. Between January 1994 through January 1999, eight patients with locally advanced malignant tumors underwent radical resection and reconstruction using pedicled VRAM flap. Inferiorly based VRAM flap was used in five patients and superiorly based VRAM flap in three patients. Defect size ranged from 144 to 900 CM2. (mean 386 cm2). Average blood loss for combined resection and reconstruction was 600ml. (range 400-800 ml.) Primary closure of both donor and recipient sites achieved in all patients. There was no wound infection, flap necrosis or abdominal hernia. There patients received postoperative radiotherapy and chemotherapy and two patients received radiotherapy only. All the the patients are alive and free of local recurrence at mean follow up 32 months. Results of our study shows that VRAM flap is versatile and sturdy flap with a wide are of rotation and it can reach diverse anatomical sites like torso, chestwall, thigh and perineum to cover large defects following radical resection for tumors. PMID- 14758884 TI - Salivary gland tumours: an analysis of 62 cases. AB - Salivary gland tumors are rare entities among the patients with head and neck neoplastic lesions. Between January 1987 to August 1997, 62 patients with salivary gland tumors presented to the surgical department of Goa Medical College, which is a tertiary referral center for the region. These patients were analyzed with an aim to study the clinical and histopathological correlation. While no age predisposition for the malignancy was found, the parotid gland was the most common site for both malignant & benign tumors (69.35%), and the minor glands (4.8%) the least common. Apart from the presence of facial nerve involvement, pain and rapid growth were also the indicators of malignancy. Fine needle aspiration cytology (FNAC) proved to be a reliable method for management of these tumors. All the patients underwent surgery with minimal morbidity and no mortality. There was no identifiable association between smoking, alcohol intake and the occurrence of the salivary glands tumors. PMID- 14758885 TI - Squamous cell carcinoma metastases to both sides of the heart--a case report. AB - We report a case, which was taken up for surgery following a diagnosis, of multicentric myxoma in both chambers of the heart, on echocardiography. Postoperative histopathological examination showed it to be squamous cell carcinoma metastases. The mode of presentation, clinical implication and rarity of this pathology are discussed. PMID- 14758886 TI - Occult breast carcinoma: a report of four cases and review of literature. AB - We report four occult carcinoma breast cases in which extensive axillary node metastases was the first manifestation. Upper outer quadrentectomy with axillary dissection was done in three patients while modified radical mastectomy was done in one. Primary tumor could be found in three patients, one had squamous cell carcinoma (SCC) & two had infiltrating duct carcinoma (IDC). However primary tumor was not detected in breast tissue of the fourth patient. Extensive lymph node metastases were found in three out of 14,15 out of 15(SCC), 24 out of 24 and 1 out of three axillary nodes respectively. Results of immunohistochemical staining for estrogen and progesterone receptors on three cases were negative. All patient received postoperative radiotherapy and chemotherapy. We have reviewed the literature and discussed the approach to diagnosis and management in female patients presenting with metastatic carcinoma in the axillary nodes with emphasis on the appropriate pre-treatment evaluation. PMID- 14758887 TI - Quantitative structure-property relationship of aromatic sulfur-containing carboxylates. AB - Based on quantum chemical calculations, TLSER model (theoretical linear solvation energy relationships) and atomic charge approach were applied to model the partition properties(water solubility and octanol/water partition coefficient) of 96 aromatic sulfur-containing carboxylates, including phenylthio, phenylsulfinyl and phenylsulfonyl carboxylates. In comparison with TLSER models, the atomic charge models are more accurate and reliable to predict the partition properties of the kind of compounds. For the atomic charge models, the molecular descriptors are molecular surface area (S), molecular shape (O), weight( MW), net charges on carboxyl group (QOC), net charges of nitrogen atoms (QN), and the most negative atomic charge (q-) of the solute molecule. For water solubility (log SW) and octanol/water partition coefficient (log KOW), the correction coefficients r2adj (adjusted for degrees of freedom) are 0.936 and 0.938, and the standard deviations are 0.364 and 0.223, respectively. PMID- 14758888 TI - Chemical forms and extractability of iron in sediments of three contrasting lakes of China and UK. AB - Iron is a limiting factor for the eutrophication of lakes, especially those lakes that are enriched with phosphorus. Extractability of iron in sediments of West Lake and Taihu Lake in China and Lower Lough Erne in Northern Ireland of UK was comparatively investigated on the basis of analysing chemical forms of iron using different extractants. It was shown that extractable iron in sediments of the lakes was greatly different using various extractants. Reactive iron or easily released iron such as "active" iron oxides, total free iron oxide and water soluble iron was not high, only accounting for 0.01%-0.15% of total iron. The efficiency of the extractants for exchangeable iron was decreased in the sequence 0.1 mol/L HCl >> DTPA + TEA mixed solution > 1 mol/L NH4OAc > 0.5 mol/L MgCl2 = 0.5 mol/L CaCl2. It seems that the complexion of iron by organic matter was not strong because the concentration of organically bound iron was significantly lower than the concentration it was forecasted. Extractable iron is not entirely consistent with or dependent on total iron in lake sediments. To a certain extent, phosphate can inhibit the release of iron in sediments of the lakes. The selection of extractants is thus the first key step to evaluate bioavailability of iron in lake sediments. PMID- 14758889 TI - An environmental approach for used oil management in Asian cities: a Bangkok' s experience. AB - This paper addresses increasing concern about the pollution threat of used oil being illegally dumped and the impact of oil on air pollution and freshwater ecosystems in Asian countries. Used oil is a very serious waste management problem. These results call for management action such as maximising the collection and recovery of used oil. The Thai government recognizes the need to recycle used oil and has been active in encouraging programs to accomplish this goal. Thus unless new approaches and incentives are developed, used oil generation may become an increasing serious problem to our environment. The purpose of this study is to examine the technical and economic feasibility, of recycling used oils. In addition, this paper briefly discusses the problems to be overcome and outlines potential mechanisms for providing the necessary disposal controls in order to maximize the protection to public health and environmental quality from potential hazards posed by used lube oil disposal. PMID- 14758890 TI - Influence of form and quantity of chromium on the development and survival of two silkworm (Bombyx mori L.) races. AB - Growth inhibitory activity has proven important in Qiufeng x Baiyu and Qingsong x Haoyue silkworm larvae. The consumption of mulberry leaves was reduced in both silkworm races while Qiufeng x Baiyu larvae showed the higher reduction in leaf consumption. From the results obtained, it was revealed that even at low concentrations of 400 mg/L of either Cr(III) or Cr(VI) ions, growth of Qiufeng x Baiyu and Qingsong x Haoyue was significantly depressed. Depression in relative growth index (RGI) and high death rate in both silkworm races indicated that the different concentrations of the two ions used caused their growth inhibitions. Through linear regression analysis, the EC50 (concentration of the compound that caused 50% reduction) was interpolated for both tested compounds. EC50 (mg/L) of Cr(III) ions in Qiufeng x Baiyu and Qingsong x Haoyue were 800 and 600 respectively. EC50 (mg/L) of Cr(VI) ions in Qiufeng x Baiyu and Qingsong x Haoyue was 600 and 316 respectively. According to the analysis of relative growth index, and an analysis of linear regression technique for measuring the growth of the silkworm races, it was indicated that the form of Cr affected growth rates, growth inhibition responses of the larvae, and toxicological effects. Thus, form and quantity of Cr accumulating in silkworms reared with contaminated leaves are likely to influence their population dynamics. PMID- 14758891 TI - Treating both wastewater and excess sludge with an innovative process. AB - The innovative process consists of biological unit for wastewater treatment and ozonation unit for excess sludge treatment. An aerobic membrane bioreactor (MBR) was used to remove organics and nitrogen, and an anaerobic reactor was added to the biological unit for the release of phosphorus contained at aerobic sludge to enhance the removal of phosphorus. For the excess sludge produced in the MBR, which was fed to ozone contact column and reacted with ozone, then the ozonated sludge was returned to the MBR for further biological treatment. Experimental results showed that this process could remove organics, nitrogen and phosphorus efficiently, and the removals for COD, NH3-N, TN and TP were 93.17%, 97.57%, 82.77% and 79.5%, respectively. Batch test indicated that the specific nitrification rate and specific denitrification rate of the MBR were 1.03 mg NH3 N/(gMLSS x h) and 0.56 mg NOx-N/(gMLSS x h), and denitrification seems to be the rate-limiting step. Under the test conditions, the sludge concentration in the MBR was kept at 5000-6000 mg/L, and the wasted sludge was ozonated at an ozone dosage of 0.10 kgO3/kgSS. During the experimental period of two months, no excess sludge was wasted, and a zero withdrawal of excess sludge was implemented. Through economic analysis, it was found that an additional ozonation operating cost for treatment of both wastewater and excess sludge was only 0.045 RMB Yuan (USD 0.0054)/m3 wastewater. PMID- 14758892 TI - Biodegradation kinetic of organic compounds of acrylic fiber wastewater in biofilm. AB - A group function relation curve between flux (J) and bulk phase concentration of substrate (S) was set up. The biodegradation kinetic of organic compounds of acrylic fiber wastewater in biofilm is studied (the treatment technology is coagulation/sedimentation-anoxic/aerobic biofilm process), and the results showed that the concentration of non-degradation pollutants in effluent is 77 mg/L. In aerobic zone, the half-rate constant is 72.84 mg/L, the maximum removal rate of organic compounds at unit area filler is very low, 0.089 g/(m2 x d), which corresponds to the fact that there are some biorefractory compounds in the wastewater. PMID- 14758893 TI - Distributions of dimethylsulfide in the Bohai sea and Yellow Sea of China. AB - Dimethylsulfide (DMS) measurements in the surface seawater of China eastern coastline were conducted during March 9-10, 1993 in Bohai Sea along the cruise from Dalian to Tianjin and during September 24-25, 1994 in Yellow Sea along the cruise from Shanghai to Qingdao. On the cruise in Bohai Sea DMS concentrations varied from 0.11 to 2.63 nmol/L with an average of 1.31 nmol/L, while DMS flux was estimated to be 0.85 micromol/(m2 x d) with the range of 0.04-3.12 micromol/(m2 x d). On the cruise in Yellow Sea DMS concentrations varied from 0.95 to 7.48 nmol/L with an average of 2.89 nmol/L, and DMS flux was estimated to be 7.94 micromol/(m2 x d) with the range of 0.11-18.88 micromol/(m2 x d). Variations in DMS concentrations along the latitude in Yellow Sea were observed larger than those along the longitude in Bohai Sea. DMS concentrations and fluxes had a similar spatial trend both in Bohai Sea and Yellow Sea with the correlation coefficients of 0.75 and 0.64, respectively. PMID- 14758894 TI - Effective incineration technology with a new-type rotary waste incinerator. AB - The technology of steady combustion in a new type of rotary incinerator is firstly discussed. The formation and control of HCl, NOx and SO2 during the incineration of sampled municipal organic solid waste are studied with the incinerator. Results showed that the new model of rotary incinerator can effectively control and reduce the pollutant formations by post combustion. PMID- 14758895 TI - Phosphorus limitation on bacterial regrowth in drinking water. AB - Assimilable organic carbon (AOC) test and bacterial regrowth potential (BRP) analysis were used to investigate the effect of phosphorus on bacterial regrowth in the drinking water that was made from some raw water taken from a reservoir located in northern China. It was shown that AOC of the drinking water samples increased by 43.9%-59.6% and BRP increased by 100%-235% when 50 microg/L PO4(3-) P(as NaH2 PO4) was added alone to the drinking water samples. This result was clear evidence of phosphorus limitation on bacteria regrowth in the drinking water. This investigation indicated the importance of phosphorus in ensuring biological stability of drinking water and offered a novel possible option to restrict microbial regrowth in drinking water distribution system by applying appropriate technologies to remove phosphorus efficiently from drinking water in China. PMID- 14758896 TI - Catalysts for decomposing ozone tail gas. AB - The preparation of immobilizing-catalysts for decomposing ozone by using dipping method was studied. XRD, XPS and TEM were used to characterize the catalysts. The three kinds of catalysts were selected preferentially, and their catalytic activities were investigated. The results showed that the catalyst with activated carbon dipping acetate (active components are Mn:Cu = 3:2, active component proportion in catalyst is 15%, calcination temperature is 200 degrees C) has the best catalytic activity for ozone decomposing. One gram of catalyst can decompose 17.6 g ozone at initial ozone concentration of 2.5 g/m3 and the residence time in reactor of 0.1 s. The experimental results also indicated that humidity of reaction system had negative effect on catalytic activity. PMID- 14758897 TI - Photodegradation of acetochlor in water and UV photoproducts identified by mass spectrometry. AB - The sunlight photodegradation half-lives of 20 mg/L acetochlor were 151, 154 and 169 days in de-ionized water, river water and paddy water, respectively. When exposed to ultraviolet (UV) light, acetochlor in aqueous solution was rapidly degraded. The half-lives were 7.1, 10.1, and 11.5 min in de-ionized water, river water and paddy water, respectively. Photoproducts of acetochlor were identified by gas chromatography/mass spectrometry (GC/MS) and found at least twelve photoproducts resulted from dechlorination with subsequent hydroxylation and cyclization processes. The chemical structures of ten photoproducts were presumed on the basis of mass spectrum interpretation and literature data. Photoproducts are identified as 2-ethyl-6-methylaniline; N,N-diethylaniline; 4,8-dimethyl-2-oxo 1,2,3,4-tetrahydroquinoline; 2-oxo-N-(2-ethyl-6-methylphenyl)-N-(ethoxymethyl) acetamide; N-(ethoxymethyl)-2'-ethyl-6'-methylformanilide; 1-hydroxyacetyl-2 ethoxyl-7-ethylindole; 8-ethyl-1-ethoxymethyl-2-oxo-1,2,3,4-tetrahydroquinoline; 4, 8-dimethyl-1-ethoxymethyl-2-oxo-1,2,3,4-tetrahydroquinoline; 2-hydroxy-2' ethyl-6'-methyl-N-(ethoxymethyl) acetanilide and a compound related to acetochlor. The other two photoproducts were detected by GC/MS although their chemical structure was unknown. PMID- 14758898 TI - Application of Neomysis awatschensis as a standard marine toxicity test organism in China. AB - The small mysid crustacean Neomysis awatschensis was collected in the west coast of Jiaozhou Bay, Qingdao, China in 1992 and acclimated and cultured in laboratory conditions since then. Standard acute toxicity tests using 4-6 d juvenile mysids of this species were conducted and the results were compared with Mysidopsis bahia, a standard toxicity test organism used in the US in terms of their sensitivities to reference toxins, as well as their taxonomy, morphology and geographic distributions. Because of its wide distribution along the Chinese coast, similar sensitivity to pollutants as M. bahia, short life history, small size and the ease of handling, this study intended to use N. awatschensis as one of the standard marine organisms for toxicity testing in China. The species were applied to acute toxicity evaluations of drilling fluid and its additives, organotin TPT and toxic algae, and to chronic (life cycle) toxicity assays of organotin TPT and a toxic dinofalgellate Alexandrium tamarense, respectively. Using N. awatschensis as a standard toxicity testing organism in marine pollution assessment in China is suggested. PMID- 14758899 TI - Effects of temperature and discharge parameters on ozone concentration of negative corona discharge. AB - The relationship of the ozone generation and the heating power, corona wire surface temperature, discharge electrode and netting electrode was studied during the negative corona discharge. The experimental results showed that the ozone concentration reduced with applied voltage decrease. With heating power increase, the ozone concentration of unit current decreased at exponential rate and it almost change no longer over 0.40 W. Under given temperature, the lower the applied voltage was, the smaller the ozone concentration was; while under given applied voltages, only over 11 kV could decrease with the surface temperature increase. The ozone concentration decreased with the lengthening of corona wire, and could reduced to 10 ppb under experimental condition of 14.2 kV; it also decreased with the shortening of wire diameter, and could decrease 67% at best in the given condition. Moreover, it decreased with the increasing size of netting electrode mesh. At the anion current of 1.65 microA, the ozone concentration of 5.0 x 5.0 cm2 is only 41% of that of 3.3 x 3.3 cm2. PMID- 14758900 TI - Water environmental planning considering the influence of non-linear characteristics. AB - In practical water environmental planning, the influence of the non-linear characteristics on the benefit of environmental investment was seldom taken into consideration. This paper demonstrates that there exist a lot of non-linear behaviors in water environment by emphatically analyzing the influence of the non linear characteristics of the economic scale, the meandering river and the model on water environmental planning, which will make a certain impact on the water environmental planning that sometimes cannot be neglected. This paper also preliminarily explores how to integrate the non-linear characteristics into water environmental planning. The results showed that compared with traditional methods, water environmental planning considering non-linear characteristics has its prevalence and it is necessary to develop the relevant planning theories and methods. PMID- 14758901 TI - To enhance the reproduction of Phanerochaete chrysosporium by adding natural lixiviums in liquid medium. AB - Great promotion to the reproduction of white rot fungus Phanerochaete chrysosporium by adding natural lixiviums such as from wood, maize core and potato in liquid medium was found in this research. Incubated in the liquid medium contained 10 mg/L glucose as carbon source with natural lixiviums for three days, the production of mycelium pellet reaches more than 80 g/L, which is 5 times more than that of without natural lixiviums. Incubated in the liquid medium contained 5 mg/L glucose as carbon source with natural lixiviums for three days, the production of mycelium pellet can reach 69.5 g/L, while the production in the medium without natural lixiviums is very low. When the liquid medium contained 1-20 g/L glucose as carbon source, the production of mycelium pellet in 3 d can only reach 12.5 g/L to 14.5 g/L. The fungus in the medium with potato lixiviums are easily contaminated by other microorganisms and in the medium with maize core lixiviums are easily bulking, while in the medium with wood lixiviums are neither easily contaminated nor bulking. Medium with wood lixiviums can produce more pellet than other medium, endure contamination and keep better sedimentation capacity. So that, wood lixivium is better additive to the culture of white rot fungi in liquid medium. Addition of the mixture of wood, maize core and potato lixiviums is of advantage to the production of mycelium pellet. The difference of the production in the medium with different amount of wood lixiviums showed little in the first 3 d, while it expanded after 3 d. Wood lixiviums stimulate the growth of P. chrysosporium instead of supply organics which fungi need. PMID- 14758902 TI - VOC removal from contaminated groundwater through membrane pervaporation. (I): Water-1,1,1-trichloroethane system. AB - 1,1,1-trichloroethane (TCA), a common chlorinated organic solvent and volatile organic compound (VOC), was removed from contaminated groundwater by using a bench-scale membrane pervaporation unit. The effects of TCA concentration, temperature, and flow rate on permeation flux and selectivity (a value) of the process were evaluated. In general, higher temperature leads to higher VOC flux but lower selectivity; higher flow rate of TCA feed stream results in higher VOC flux and selectivity, an indication of the effect of concentration polarization; higher TCA feed concentration produces higher TCA permeation, however, the selectivity was virtually unchanged. PMID- 14758903 TI - VOC removal from contaminated groundwater through membrane pervaporation. (II): 1,1,1-trichloroethane-SDS surfactant solution system. AB - The conventional "pump-and-treat" technology for subsurface remediation of groundwater contaminated with volatile organic compounds (VOCs) such as 1,1,1 trichloroethane (TCA), a common chlorinated organic solvent, has limitation of prohibitively long treatment time due to extremely low water solubility of the VOCs. Surfactant-based soil remediation has emerged as the effective technology that substantially reduces the treatment time. In order to make the whole process economical, the surfactant used in soil washing has to be recovered and reused. This study examined the recovery of anionic surfactant, sodium dodecyl sulfate (SDS), from soil remediation fluids containing TCA, using a bench-scale membrane pervaporation unit. The effects of high TCA concentration, surfactant dosage, and flow rate on permeation flux and selectivity (alpha value) of the process were evaluated. In general, higher surfactant concentration yielded lower TCA flux and constant water flux, resulting in declining a values; higher flow rate of TCA feed stream results in higher VOC flux and selectivity, an indication of the effect of concentration polarization; higher TCA feed concentration produces higher TCA permeation across the membrane, however, the selectivity was virtually unchanged unless the total TCA concentration exceeded 2000 ppm. PMID- 14758904 TI - A simulation analysis of the migration and transformation of pollutants contained in landfill leachate. AB - A dynamic composite model for a soil-water system that can be used to simulate the movement of leachate from a landfill. The composite model includes nine sub models that trace water movement and the migration and transformation of five pollutants (organic N, NH4-, NO3-, NO2-, and Cl-) in saturated and unsaturated soil. The model to simulate the movement of leachate from a landfill in Laogang Town, Shanghai City was used. In this application, the values for the model parameters were obtained by performing a laboratory simulation experiment of water movement and pollutant migration and transformation in soil columns. Soil and leachate obtained from the landfill site and its vicinity were used in the laboratory experiments. The model was then used to simulate leachate movement and pollutant activity during the ten-year period when the landfill was in operation and in the twenty-year period following its closure. The simulation results revealed that the leachate migrated into the groundwater at the rate of 90-100 meters per year. This model can be applied in the design of future landfills in China for the purpose of assessing and forecasting leachate plumes. PMID- 14758905 TI - Elemental sulfur effects on Pb and Zn uptake by Indian mustard and winter wheat. AB - A pot experiment was conducted to investigate the influence of elemental sulfur to contaminated soil on plant uptake by a heavy metal hyperaccumulator, Indian mustard (Brassica juncea) and a field crop, winter wheat (Triticum. aestivum). Elemental sulfur (S) with different rates was carried out, they were 0 (S0), 20 (S20), 40 (S40), 80 (S80), and 160 (S160) mmol/kg respectively. Extra pots with the same rates of S but without plants were used for soil sampling to monitor pH and CaCl2-extractable heavy metal changes. The results showed that S enhanced phytoextraction of Pb and Zn from contaminated soil. Application S effectively decreased soil pH down to 1.1 as the most at the rate of S160. The concentrations of CaCl2-extractable Pb and Zn in soil and uptake of Pb and Zn by the plants were increased with soil pH decreased. A good correlation between CaCl2-extractable Pb/Zn and soil pH was found (R2Pb = 0.847 and R2Zn = 0.991, n = 25). With S application, soil CaCl2-extractable Pb and Zn concentrations, concentration of Pb and Zn in plants and the amount of removal by plant uptake were significantly higher than those without S. Under the treatment of S160, the highest CaCl2 extractable Pb and Zn were observed, they were 4.23 mg/kg and 0.40 mg/kg, 2.7 and 2.0 times as that of the control (S0) respectively. At the highest rates of S (S160), both Indian mustard and winter wheat reached the highest uptake of Pb and Zn. The highest Pb concentrations in wheat and Indian mustard were 32.8 mg/kg and 537.0 mg/kg, all 1.8 times as that of the control, and the highest Zn concentrations in wheat and Indian mustard were 215.5 mg/kg and 404.0 mg/kg, 2.4 and 2.0 times as that of the control respectively. The highest removals of Pb and Zn from the contaminated soil were 0.41 mg/pot and 0.31 mg/pot by Indian mustard in the treatment of S160 through 50 days growth. PMID- 14758906 TI - Changes in phototactic behavior of Daphnia magna clone C1 242 in response to copper, cadmium and pentachlorophenol. AB - In order to develop a sound biotechnique for monitoring water quality that builds on the previous experiments carried out in our laboratory, a specific D. magna clone C1 242 was used to study the effects of pollutants on phototactic behavior. In all experiments, the animals showed a stable and repeatable phototactic index approximated 0.2 in the presence and 0.4 in the absence of fish kairomones, which decreased significantly in response to pollutants. There existed no pollutant x fish kairomone interaction, indicating the changes in phototactic behavior of animals imposed by pollutants were independent of the presence of fish kairomones. The detection limits for changes in phototactic behavior of D. magna clone C1 242 are 0.04 mg/L for copper, 0.02 mg/L for cadmium, and 0.80 mg/L for PCP, respectively, quite lower than LC50 (48 h). The changes in phototactic behavior in presence to pollutants occurred quickly (3 h) compared to the period over whole acute toxicity tests. Therefore, D. magna clone C1 242 could be potentially used to monitor water quality. Moreover, the phototactic behavior did not decrease further in the pollutant mixtures employed in our experiments compared to individual pollutants, except in the Cd-PCP treatment. This fact suggests that the formation of water quality criteria must be based upon pollutant mixture tests. PMID- 14758907 TI - Polycyclic aromatic hydrocarbon in rain and street runoff in Amman, Jordan. AB - Concentrations of polycyclic aromatic hydrocarbons (PAHs) were determined in the rain and street runoff samples from two sites in the vicinity of Amman City during the pluvial period 1999-2000. The results showed that elevated levels of PAHs were detected in the city center (site 1) than the residential area (site 2) and that the levels were higher in street runoff than rain samples of the same sites. The highest concentration of PAHs in both street runoff and rain samples were observed in the first rainy month (November 1999) which indicated a wash out effect of PAHs originating from vehicular emission accumulated during the long dry summer season before sampling. Within the investigated cold winter seasons, fluctuations in PAHs concentration were observed. The variation was attributed to the fossil combustion for heating purposes and to intervals between rainfalls: as the longer the intervals between rains were, the higher the PAH concentration were. Removal of PAHs from the atmosphere through precipitation over the investigated period varied with time and places depending on the amount of rainfall where higher rainfall removed higher amount of PAHs from the atmosphere. The amount of PAHs washed out through precipitation was estimated to be around 14.8 mg/m2 and 21.1 mg/m2 for sites 1 and 2 respectively. PMID- 14758908 TI - Rapid ecotoxicological assessment of heavy metal combined polluted soil using canonical analysis. AB - Quick, simple to perform, and cheap biomarkers were combined in a rapid assessment approach to measure the effects of metal pollutants, Cu, Cd, Pb and Zn in meadow burozem on wheat. Analysis of orthogonal design showed that the significant zinc factor indicated both the inhibition rate of shoot mass and that of root elongation were affected by zinc (P < 0.05 and P < 0.01, respectively). The first toxicity canonical variable (TOXI), formed from the toxicity data set, explained 49% of the total variance in the toxicity data set; the first biological canonical variable (BIOL) explained 42% of the total variation in the biological data set. The correlation between the first canonical variables TOXI and BIOL (canonical correlation) was 0.94 (P < 0.0001). Therefore, it is reliable and feasible to use the achievement to assess toxicity of heavy metal combined polluted soil using canonical analysis. Toxicity of soil combined polluted by heavy metals to plant community was estimated by comparing the IC50 values describing the concentration needed to cause 50% decrease with grow rate compared to no metal addition. Environmental quality standard for soils prescribe that all these tested concentration of heavy metals in soil should not cause hazard and pollution ultimately, whereas it indicated that the soils in second grade cause more or less than 50% inhibition rates of wheat growth. So environmental quality standard for soils can be modified to include other features. PMID- 14758909 TI - Influence of nonionic surfactant on the solubilization and biodegradation of phenanthrene. AB - Phenanthrene was solubilized in two different nonionic surfactants, Tween80 and Triton X-100. The bioavailability of phenanthrene to the bacteria isolated from the petroleum contaminated soils was studied based on the rotary flasks experiments. The results showed that the concentration of nonionic surfactants above the critical micelle concentration (CMC) can increase the solubility of phenanthrene in water and were innoxious to the phenanthrene-degrading bacteria; phenanthrene solubilized in the micelles of Tween80 was bioavailable and biodegradable. The research demonstrated the potential of surfactant-enhanced bioremediation of soils contaminated by hydrophobic organic compounds (HOCs). PMID- 14758910 TI - Development of thermochromic strips as a water pasteurization indicator. AB - Boiling of water, for purification, commonly practiced in the world, has many problems associated with it like danger of scalding, scaling in the vessels, removal of useful minerals and blandness of taste etc. Water can be made safe for drinking simply by heating at 65 degrees C for 6 minutes. A colour indicating strip was developed which changes colour from red to purple at 67 degrees C. Use of this strip can help in pasteurizing water without the above problems and with considerable energy saving. PMID- 14758911 TI - Plastic surgery. PMID- 14758912 TI - Tissue engineering--body parts from the Petri dish. AB - The development of methods to regenerate human tissues and organs by tissue engineering (TE), will have a dramatic influence on many medical specialities in the future. The essence of plastic surgery is to reconstruct disrupted and damaged tissues by the use of a plethora of techniques spanning from small local skin flaps to highly advanced microsurgery and free composite grafts. However, these methods only focus on moving tissue from one part of the patient to another without actual regeneration. To be able to take the next step in development of the speciality it is of necessity to address this issue. Hence it follows naturally that plastic surgeons lead and represent the driving force of the development within the research of tissue engineering. In this paper we would like to present active research and also give an overview of areas in tissue engineering that are of special interest to the plastic surgeon. PMID- 14758913 TI - Immediate breast reconstruction. AB - Immediate breast reconstruction (IBR) in conjunction with mastectomy for cancer or high risk of breast cancer is safe from an oncological point of view. The cosmetic outcome can be excellent, especially when performing mastectomy by sparing the skin of the breast and reconstructing the breast mound with autogenous tissue. The majority of women at their working age are willing to have their breast reconstructed. Patients with newly diagnosed cancer undergoing major surgery need extra support compared with those undergoing late reconstructions. Immediate reconstructions with one operation, one hospital stay and one sick leave are economically favourable by diminishing the demand of delayed reconstructions. Preference in patient selection for IBR should be in patients with good prognoses like those with diffuse non-invasive cancer and those with a considerable risk to develop breast cancer. Women with axillary-node negative invasive cancer and women with late local recurrences in a breast earlier conservatively treated are also suitable for IBR. If needed, oncological treatments can be given after IBR, although radiotherapy after pure implant reconstructions is not recommended. In order to give all eligible patients an equal opportunity to have IBR, treatment of breast patients should be centralised to hospitals with a team comprising breast cancer surgeons, pathologists, radiologists, and plastic surgeons. PMID- 14758914 TI - Soft tissue sarcoma of the trunk and extremities. AB - The diagnostic methods and treatment modalities of soft tissue sarcomas have evolved with the multidisciplinary approach. The soft tissue sarcoma team must have specialists capable of using and combining modern methods of radiology and pathology, cytogenetics, tumour surgery, tissue transfer techniques, radiotherapy and chemotherapy for optimal local and systemic treatment. Limb sparing surgery combined with radiotherapy has lowered the amputation rate and maintained low rates of local recurrence. Reconstructive surgery facilitates treatment of patients with soft tissue sarcoma by permitting tumour resection with adequate margins, protects vital structures, enables early postoperative radiation therapy, maintains extremity length, and if necessary assists in palliative procedures. The ability to maintain function and aesthetics after tumour resection, and effective palliation improves the quality of life for these patients. Early recognition and appropriate referral to a tumour centre improve the outcome. PMID- 14758915 TI - Soft-tissue and bone reconstruction in compound fractures of the lower leg. PMID- 14758916 TI - Cleft lip and palate surgery. PMID- 14758917 TI - Craniofacial surgery over 30 years in Goteborg. AB - Craniofacial surgery is a new sub-specialty in the field of plastic reconstructive surgery and is dedicated to the treatment of severe cranial and facial malformations. Craniofacial surgery gradually started in Goteborg in the late 1970's and has been acting as the Scandinavian center since 1983. Over these 30 years an almost complete change in surgical techniques has evolved. Also profound changes in timing of surgery have followed. Results have been dramatically improved based on critical evaluation of standardized registration of long-term results. One of the most dramatic developments has been the introduction of implantable stainless steel springs. This has changed the treatment of Craniosynostosis completely and has made midfacial advancement procedures possible without relapse. PMID- 14758918 TI - Perspective on contemporary burn surgery and burn care in Sweden. AB - Burn surgery has gone through revolutionary changes during the last century, even the most severe burns can now be safely treated. Modern surgical treatments have been followed by improvements in intensive burn care and anaesthesiology. These developments have allowed immediate surgery within 24 hours of the injury to take place in our centre, therefore obtaining advantageous results in shortening hospital stay, improving patient's functional results and simplifying the treatment itself. This treatment is economically beneficial as well. In future we count on rapid development of a new discipline, tissue engineering, which should take a greater role in burn care. PMID- 14758919 TI - Liposuction in arm lymphedema treatment. AB - Breast cancer is the most common disease in women, and up to 38% develop lymphedema of the arm following mastectomy, standard axillary node dissection and postoperative irradiation. Limb reductions have been reported utilizing various conservative therapies such as manual lymph and pressure therapy. Some patients with long-standing pronounced lymphedema do not respond to these conservative treatments because slow or absent lymph flow causes the formation of excess subcutaneous adipose tissue. Previous surgical regimes utilizing bridging procedures, total excision with skin grafting or reduction plasty seldom achieved acceptable cosmetic and functional results. Microsurgical reconstruction involving lympho-venous shunts or transplantation of lymph vessels has also been investigated. Although attractive in concept, the common failure of microsurgery to provide complete reduction is due to the persistence of newly formed subcutaneous adipose tissue, which is not removed in patients with chronic non pitting lymphedema. Liposuction removes the hypertrophied adipose tissue and is a prerequisite to achieve complete reduction. The new equilibrium is maintained through constant (24-hour) use of compression garments postoperatively. Long-term follow up (7 years) does not show any recurrence of the edema. PMID- 14758920 TI - Abdominal wall reconstruction with vascularised autologous tissue. PMID- 14758921 TI - In vitro studies of erythrocyte-vascular endothelium interactions. AB - Abnormal interactions between red blood cells (RBCs) and vascular endothelial cells (ECs) are crucial factors in causing vascular pathology in several diseases, including vaso-occlusive crises in sickle cell anemia and the development of vascular complications in diabetes mellitus and malaria. A mechanistic understanding of the specific nature of RBC-EC interactions and ensuing functional consequences can provide insights into the pathophysiology of RBC-related vascular disorders and a rational basis for developing novel therapies. This review discusses in vitro experimental models that are commonly used for investigating RBC-EC interactions, and current knowledge of the molecular mechanisms of RBC-EC adhesion and EC functions modulated by RBCs. Because blood flow-induced mechanical forces and convective mass transfer play significant roles in regulating vascular events, it is necessary to develop advanced dynamic experimental models for elucidating RBC-EC interactions under well-controlled, physiologically relevant mechanical environments. PMID- 14758922 TI - Protofilament and hexagon: a three-dimensional mechanical model for the junctional complex in the erythrocyte membrane skeleton. AB - It is a long-standing mystery why erythrocyte actin filaments in the junctional complex (JC) are uniformly approximately 37 nm and the membrane skeleton consists of hexagons. We have previously proposed that a "molecular ruler" formed by E tropomodulin and tropomyosin 5 or 5b functions to generate protofilaments of 12 G actin under mechanical stress. Here, we illustrate that intrinsic properties of actin filaments, e.g., turns, chemical bonds, and dimensions of the helix, also favor fragmentation into protofilaments under mechanical stress. We further construct a mechanical model in that a pair of G actin is wrapped around by a split alpha and beta spectrin, which may spin to two potential positions, and stabilize to one when the tail end is restricted. A reinforced protofilament may function as a mechanical axis to anchor three (top, middle, and bottom) pairs of Sp. Each Sp pair may wrap around the protofilament with a wide dihedral angle (approximately 166.2 degrees) and a minimal axial distance (2.75 nm). Such three Sp pairs may spiral down (right handed) the protofilament from the pointed end with a dihedral angle of approximately 55.4 degrees in between the Sp pairs. This first three-dimensional model of JC may explain the hexagonal geometry of the erythrocyte membrane skeleton. PMID- 14758923 TI - Mechanical asymmetry in the embryonic chick heart during looping. AB - Cardiac looping, which begins with ventral bending and rightward rotation of the primitive heart tube, is an essential morphogenetic event that occurs early in vertebrate development. The biophysical mechanism that drives this process is unknown. It has been speculated that increased stiffness along the dorsal side of the ventricle combined with an intrinsic cardiac force causes the heart to bend. There is no experimental support for this hypothesis, however, since little is known about regional mechanical properties of the heart during looping. We directly measured diastolic stiffness of the inner curvature (IC), outer curvature (OC), and dorsal-ventral sides of the stage 12 chick heart by microindentation. The IC of intact hearts was found to be significantly stiffer than either the OC or the sides. which were of similar stiffness. Isolated cardiac jelly, which is a thick, extracellular matrix compartment underlying the myocardium, was approximately an order of magnitude softer than intact hearts. The results of a computational model simulating the indentation experiments, combined with the stiffness measurements, suggests the regional variation in stiffness is due to the material properties of the myocardium. A second model shows that a relatively stiff IC may facilitate bending of the heart tube during looping. PMID- 14758924 TI - Effects of periodic body acceleration on the in vivo vasoactive response to N omega-nitro-L-arginine and the in vitro nitric oxide production. AB - Periodic acceleration (pGz), a novel method of ventilatory support, is achieved using a platform that moves cyclically in the headward-footward direction. PGz has been shown to increase vascular shear stress and regional blood flows, as well as decrease pulmonary and systemic vascular resistances. PGz also increases nitric oxide (NO) production. This study was undertaken to determine the effects of pGz on the NO inhibiting effects of N-w-nitro-L-arginine (L-NAME) in vivo, and to determine if increased NO production due to pGz could be reproduced in vitro with isolated arteries. Pigs were assigned to conventional ventilation (CV), or pGz, with no additional breathing assistance. L-NAME was infused in cumulative doses of 1, 3, 10, 30, and 100 mg/kg. Cardiac output decreased in both groups by 50%. There was also a dose-dependent increase in blood pressure, pulmonary artery pressure, and vascular resistances. However, pGz attenuated the vascular response of L-NAME. Isolated porcine aortas exposed to nonpulsatile, pulsatile, and pulsatile flow plus pGz exhibited an increase in nitrites with the addition of pulsatile flow (300%, relative to steady flow), and a further increase with pGz (1000%, relative to steady flow). It has been determined that pGz, a novel method of increasing shear stress on the vascular endothelium. attenuates the vasoactive response to L-NAME. The in vitro experiments demonstrated that increases in NO production in vivo could be reproduced in vitro, which provides the opportunity to investigate the mechanisms of cardiovascular pGz effects. PMID- 14758925 TI - A device for long term, in vitro loading of three-dimensional natural and engineered tissues. AB - In vitro studies of mechanical loads applied to three-dimensional tissue constructs are important to the design and production of functional, engineered bone tissue. This study reports the development and characterization of a mechanical device capable of subjecting a three-dimensional section of natural or engineered tissue to precise, reproducible four-point bending deformations over a range of programmable magnitudes and frequencies. To test the biological and mechanical capabilities of the system, a low-cycle (360 cycles/day), medium-range strain (2500 microstrain), long-term (16 day) loading regime was applied to rat bone marrow stromal cells cultured in porous DL-polylactic acid scaffolds. Cells proliferated in culture throughout the experiment, and with time showed an increase in alkaline phosphatase expression per cell. Calcium and phosphorus mineral deposition by the unloaded group was significantly greater (p<0.05) than that deposited by the loaded group. The molar ratio of calcium to phosphorus in the unloaded group (0.94:1) was significantly greater (p<0.05) than that of the loaded group (0.41:1). The loading device presented here is a tool which can be used to help elucidate contributions of mechanical loading/fatigue on biodegradable materials, as well as study the effects of mechanical loading on natural or engineered tissues in vitro. PMID- 14758926 TI - Compression induced cell damage in engineered muscle tissue: an in vitro model to study pressure ulcer aetiology. AB - The aetiology of pressure ulcers is poorly understood. The complexity of the problem, involving mechanical, biochemical, and physiological factors demands the need for simpler model systems that can be used to investigate the relative contribution of these factors, while controlling others. Therefore, an in vitro model system of engineered skeletal muscle tissue constructs was developed. With this model system, the relationship between compressive tissue straining and cell damage initiation was investigated under well-defined environmental conditions. Compression of the engineered muscle tissue constructs revealed that cell death occurs within 1-2 h at clinically relevant straining percentages and that higher strains led to earlier damage initiation. In addition, the uniform distribution of dead cells throughout the constructs suggested that sustained deformation of the cells was the principle cause of cell death. Therefore, it is hypothetised that sustained cell deformation is an additional mechanism that plays a role in the development of pressure ulcers. PMID- 14758927 TI - Age-related changes of noncalcified collagen in human cortical bone. AB - It is well known that osteoid, which contains a noncalcified collagenous matrix, is formed during the initial stage of bone formation during the bone remodeling process. Thus, synthesis of defective collagen molecules in osteoid may cause abnormal bone formation, thereby leading to changes in bone quality. The objective of this study was to investigate age-related changes in noncalcified collagen molecules in osteoid and its likely effects on the mechanical integrity of human cortical bone. Thirty human cadaveric femurs were divided into three age groups: young adults, middle age, and the elderly, respectively. A novel high performance liquid chromatography approach was employed to quantify the denaturation of noncalcified collagenous matrix in addition to mechanical tests of bone. Bulk concentrations of both enzymatic and nonenzymatic collagen cross links in bone also were measured. Moreover, the number of newly formed osteons per unit area and the bony area fraction of these osteons were estimated. The results of this study indicate that denaturation of the noncalcified collagenous matrix in bone increases with increasing age. In addition, such collagen denaturation in osteoid exhibited a correlation with nonenzymatic collagen cross links as well as the strength and toughness of bone. These results suggest that age-related changes in the noncalcified collagenous matrix induced by bone remodeling may have likely effects on bone quality. PMID- 14758928 TI - Measuring forces in liver cutting: new equipment and experimental results. AB - We are interested in modeling the liver cutting process as accurately as possible by determining the mechanical properties experimentally and developing a predictive model that is self-consistent with the experimentally determined properties. In this paper, we present the newly developed hardware and software to characterize the mechanical response of pig liver during (ex vivo) cutting. We describe the custom-made cutting apparatus, the data acquisition system, and the characteristics of the cutting force versus displacement plot. The force displacement behavior appears to reveal that the cutting process consists of a sequence of intermittent localized crack extension in the tissue on the macroscopic scale. The macroscopic cutting force-displacement curve shows repeating self-similar units of localized linear loading followed by sudden unloading. The sudden unloading coincides with observed onset of localized crack growth. This experimental data were used to determine the self-consistent local effective Young's modulus for the specimens, to be used in finite element models. Results from finite element analyses models reveal that the magnitude of the self consistent local effective Young's modulus determined by plane-stress and plane strain varies within close bounds. Finally, we have also observed that the local effective Young's modulus determined by plane stress and plane strain analysis decreases with increasing cutting speed. PMID- 14758929 TI - Rational design of contact guiding, neurotrophic matrices for peripheral nerve regeneration. AB - Nerve guides filled with magnetically aligned hydrated gels of type I collagen have been shown to impart strong contact guidance cues to elongating neurites in vitro and to increase the number of regenerating axons in vivo relative to an isotropic collagen gel. We have formulated and analyzed a model to determine the conditions under which the target concentration of nerve growth factor (NGF) to support axonal growth can be sustained by entrapping either NGF-secreting cells or NGF-releasing polymer microspheres in the aligned gel. The equation describing NGF concentration with a distributed source term was solved after experimental determination of (1) the rate of NGF release from PLGA 85/15 microspheres, (2) the NGF diffusion coefficient in the gel and nerve guide membrane containing the gel, and (3) the maximum microsphere loading that does not compromise the magnetic alignment of collagen fibrils. We find that for a rat sciatic nerve, when using a 1 mm diameter nerve guide within a commercially available collagen membrane, the microsphere loading limit will prevent the construct's capacity to sustain the target NGF concentration of 1 ng/ml at two months when either wild type Schwann cells or PLGA 85/15 microspheres are used as the NGF source. This target concentration, however, will be maintained when transfected cells described in the literature to hypersecrete NGF are used, or when the microspheres are used if the permeability of the nerve guide membrane can be moderately decreased. For a human median nerve, when using a 5 mm diameter nerve guide within a commercially available membrane, the microspheres are capable of sustaining NGF concentrations above 1 ng/ml to at least 75 days without the need to decrease membrane permeability. PMID- 14758930 TI - Modeling soluble gas exchange in the airways and alveoli. AB - A mathematical model of heat, water and soluble gas exchange in the airways and alveoli was used to predict the location of soluble gas exchange in the lung. A previously published model of heat, water and soluble gas exchange in the airways was improved by incorporating anatomical data on the airway wall to better describe the bronchial circulation and expanding the model to include a time varying description of soluble gas concentration in the alveoli. Next, the model was validated using two experimental data sets from the literature: (1) ethanol expirograms and (2) the uptake of seven soluble gases. Then, the model simulated the excretion of ten soluble gases whose blood:air partition coefficient (lambda(b:a)), a measure of blood solubility, ranged over 5 orders of magnitude. We found that gases with lambda(b:a) < 10 exchange almost solely in the alveoli and gases with lambda(b:a) > 100 exchange almost exclusively in the airways. Gases with lambda(b:a) between 10 and 100 have significant interaction with the airways and alveoli. These results suggest that the airways play a larger role in pulmonary gas exchange than previously assumed and may require a reevaluation of pulmonary tests that involve exhaled samples of gases with lambda(b:a) > 10. PMID- 14758931 TI - Estimation of delay times in biological systems. AB - The problem of delay time estimation in biological systems is addressed with the focus on practical applicability of methods. Four delay time estimators are described: a cross correlation method and three increasingly sophisticated interpretations of the phase spectrum, ranging from a pointwise interpretation of the phase spectrum in terms of a delay to a Hilbert transform method. The four methods are compared through simulation studies showing that, in general, the Hilbert transform method performs best. The methods are then used to estimate delay times in three physiological systems: vestibular stimulation, cerebral autoregulation, and human orthostatic tremor. In all three cases, the Hilbert transform method yields the best results, leading in some cases to physiologically more sensible interpretations of experiments than the other methods. PMID- 14758932 TI - Skin hemangiomas and occult dysraphism. PMID- 14758933 TI - Volume-outcome. PMID- 14758934 TI - Isolated flat capillary midline lumbosacral hemangiomas as indicators of occult spinal dysraphism. AB - OBJECT: Historically, cutaneous stigmata representative of occult spinal dysraphism (OSD) have included lumbar hemangiomas. Frequently, this skin change is found in conjunction with other cutaneous alterations such as dermal sinus tracts and subcutaneous lipomas. Debate has recently surrounded the question of whether these skin changes in isolation might indicate underlying spinal disease. The authors reviewed their experience in their most recent 120 cases in which OSD was diagnosed. METHODS: The authors retrospectively reviewed records obtained in 120 patients with OSD. They found that many of the patients reviewed harbored only a flat capillary hemangioma as an indicator of OSD. In 21 patients (17.5%) with only midline lumbosacral flat capillary hemangiomas, underlying OSD was present. No single variety of OSD had a higher incidence of association with this single cutaneous stigma. CONCLUSIONS: Based on their experience, the authors recommend magnetic resonance (MR) imaging evaluation in cases involving this skin lesion in isolation to discern the potential for surgically significant spinal cord anomalies. Prospective studies are now needed to examine MR images obtained in all children with this lesion in the midline lumbosacral spine and assess for OSD. PMID- 14758935 TI - In-hospital mortality rates after ventriculoperitoneal shunt procedures in the United States, 1998 to 2000: relation to hospital and surgeon volume of care. AB - OBJECT: Death after ventriculoperitoneal (VP) shunt surgery is uncommon, and therefore it has been difficult to study. The authors used a population-based national hospital discharge database to examine the relationship between annual hospital and surgeon volume of VP shunt surgery in pediatric patients and in hospital mortality rates. METHODS: All children in the Nationwide Inpatient Sample (1998-2000, age 90 days-18 years) who underwent VP shunt placement or shunt revision as the principal procedure were included. Main outcome measures were in-hospital mortality rates, length of stay (LOS), and total hospital charges. Overall, 5955 admissions were analyzed (253 hospitals, 411 surgeons). Mortality rates were lower at high-volume centers and for high-volume surgeons. In terms of hospital volume, the mortality rate was 0.8% at lowest-quartile volume centers (< 28 admissions/year) and 0.3% at highest-quartile-volume centers (> 121 admissions/year). In terms of surgeon volume, the mortality rate was 0.8% for lowest-quartile-volume providers (< nine admissions/year) and 0.1% for highest-quartile-volume providers (> 65 admissions/year). After multivariate adjustment for demographic variables, emergency admission and presence of infection, hospital volume of care remained a significant predictor of death (odds ratio [OR] for a 10-fold increase in caseload 0.38; 95% confidence interval [CI] 0.18-0.81). Surgeon volume of care was statistically significant in a similar multivariate model (OR for a 10-fold increase in caseload 0.3; 95% CI 0.13-0.69). Length of stay was slightly shorter and total hospital charges were slightly higher at higher-volume centers, but the differences were not statistically significant. CONCLUSIONS: Pediatric shunt procedures performed at high-volume hospitals or by high-volume surgeons were associated with lower in hospital mortality rates, with no significant difference in LOS or hospital charges. PMID- 14758936 TI - Ineffectiveness of dietary folic acid supplementation on the incidence of lipomyelomeningocele: pathogenetic implications. AB - OBJECT: Periconceptual folic acid supplementation is effective in myelomeningocele prevention. The relationship between folic acid and lipomyelomeningocele (LMM) and the overall incidence of this occult form of spina bifida has never been studied. The objectives of this study were to determine the impact of dietary folic acid supplementation on the incidence of LMM and to measure its overall incidence. METHODS: In a retrospective population-based study the authors calculated the incidence of LMM in Nova Scotia between 1985 and 2001. Because of changes in public policy during this period, there are three intervals defined in relation to the treatment of the food supply with folic acid: 1) prior to folic acid fortification (1985-1994); 2) postsupplementation but prefortification (1995-1998); and 3) postfortification. The overall incidence of LMM in Nova Scotia between 1985 and 2001 was 16 per 100,000 live births or one case per 6121 live births. Its incidence between 1985 and 1994 was 15 per 100,000 live births, and between 1995 and 1998 it was 12 per 100.000 live births (relative risk [RR] 0.82, 95% confidence interval [CI] 0.31-2.22; p = 0.7). Between 1999 and 2001, the incidence of LMM was 29 per 100,000 live births, which was not significantly different from that between 1995 and 1998 (RR 2.41. 95% CI 0.79-7.36; p = 0.11) or between 1985 and 1994 (RR 1.98, 95% CI 0.86-4.56; p = 0.1). CONCLUSIONS: The overall incidence of LMM between 1985 and 2001 in Nova Scotia was 16 per 100,000 live births and has not been reduced by dietary folic acid supplementation. This finding provides epidemiological evidence that the embryogenesis of LMM is fundamentally different from that of myelomeningocele. PMID- 14758937 TI - Syringomyelia complicating myelomeningocele: review of the evidence. AB - Syringomyelia is frequently found in association with myelomeningocele. Although often asymptomatic, it can, in exceptional cases, cause severe morbidity. The author performed a structured literature review to address the following aspects of this clinical problem. What is the natural history? There are data concerning the imaging prevalence, the autopsy prevalence, and the prevalence of clinically active syringomyelia among patients with myelomeningocele, but literature provides no description of the course of this condition over time. What is the clinical significance? That correlations among symptoms, signs, treatments, and imaging findings are poorly described is no surprise in view of the large fraction of patients with syringomyelia who are asymptomatic. There is, however, an impressive mass of anecdotal evidence associating progressive syringomyelia with untreated (or inadequately treated) hydrocephalus. How to make the diagnosis? There is no disagreement that magnetic resonance imaging is the diagnostic modality of choice, but the literature provides very little guidance about who should undergo this investigation. There is no analysis of the costs and benefits of screening or periodic surveillance. What is the best treatment? Many surgical procedures have been reported to be useful as components of complex algorithms relating clinical factors and imaging data to treatment recommendations. There have been no controlled studies. There have been no prospective studies. There have been no multiinstitutional studies. No studies in which outcomes have been evaluated using objective, validated instruments. No studies in which the robustness of treatment effects over time have been documented. The complexity of treatment selection in contemporary practice makes syringomyelia unsuitable for randomized controlled trials of different therapies. Prospective cohort studies are feasible, however, and hold the potential to address many important questions about natural history and patient outcomes. PMID- 14758938 TI - Cortical dysplastic lesions in children with intractable epilepsy: role of complete resection. AB - OBJECT: The authors conducted a study to determine seizure-related outcomes in a group of pediatric patients with pathologically proven focal cortical dysplasia (FCD) treated by focal cortical resections and multiple subpial transections (MSTs). METHODS: The authors performed a retrospective review of pediatric patients in whom surgery was conducted to treat medically refractory epilepsy secondary to cortical dysplasia between April 1989 and January 2001. Diagnostic studies included preoperative scalp electroencephalography (EEG), magnetic resonance (MR) imaging, positron emission tomography (PET), and magnetoencephalography (MEG). Intraoperative electrocorticography (ECoG) or extraoperative subdural grid EEG monitoring was performed in all patients. Seizure outcome was classified using the Engel scheme. The authors analyzed nine data points and compared these with seizure outcome, including seizure semiology, MR imaging, PET and MEG data, as well as location of resection, intracranial video-EEG findings, MSTs, postresection ECoG data, and histological findings. The authors analyzed data obtained in 39 children in whom the follow-up interval after epilepsy surgery was at least 18 months. Patients had suffered epilepsy for a mean of 7.7 years prior to surgical intervention and their mean age at treatment was 9.6 years (range 2 months-18 years). A good seizure-related outcome was demonstrated in 28 patients (72%), including 21 (54%) who were free of seizures (Engel Class I) and seven (18%) in whom seizures were rare (Engel Class II). In 11 patients seizure-related outcome was less favorable, including six (15%) with worthwhile improvement involving some seizures (Engel Class III) and five (13%) with no postoperative seizure improvement (Engel Class IV). There was no significant correlation between seizure outcome and data related to seizure characteristics, MR imaging, PET scanning, MEG, location of resection, intracranial video-EEG, postresection ECoG, and histological findings. Eight (50%) of 16 patients who underwent MSTs in addition to incomplete resection of FCD experienced a good outcome (Engel Class I and II). Twenty (87%) of 23 patients in whom resection of FCD was complete and in whom MSTs were not performed experienced a good seizure outcome (p < 0.05). CONCLUSIONS: Complete resection of FCD results in good seizure outcome in a majority of children. When conducted in conjunction with incomplete cortical resection, MSTs do not improve seizure outcome in patients with FCD. Focal cortical dysplasia located outside of eloquent cortex and complete excision of the lesion are the most important predictors of seizure outcome. PMID- 14758939 TI - Multiple subpial transections in the treatment of pediatric epilepsy. AB - OBJECT: The technique involved in multiple subpial transections (MSTs) allows the surgeon treating patients with epilepsy the capability to make disconnective lesions in epileptogenic regions of eloquent cortex. Although there have been increasing numbers of reports in adults of the efficacy and relative safety of this technique, there are relatively few such reports in children. The authors present their experience in 30 children who underwent MSTs during the surgical management of the seizure disorder. METHODS: Thirty consecutive children who underwent MSTs with or without cortical excision form the basis of this retrospective review. An analysis of neurological adverse effects following MSTs and seizure outcome was performed. Between 1996 and 2000, MSTs were performed either as stand-alone therapy (four patients) or in conjunction with planned cortical excisions (26 patients). Twenty-three children underwent invasive monitoring after placement of subdural grid electrodes, and in seven intraoperative electrocorticography alone was performed. The mean follow-up period for the group was 3.5 years (minimum 30 months in all cases). All 20 patients in whom MSTs were performed in the primary motor cortex experienced transient hemiparesis (mild in 12 and moderate in eight) lasting up to 6 weeks; however, no patient suffered a permanent motor deficit in the long-term follow-up period. In 26 patients who underwent cortical resections followed by MSTs, 12 (46%) were seizure free (Engel Class I) following surgery. Eleven patients (42%) (Engel Classes II and III) continued to suffer seizures but improvement in seizure control was adequate following surgery. In the 23 patients in whom subdural grids were placed to capture the ictal onset zone by invasive video electroencephalography, MSTs comprised a mean of 37% of the surgically treated area under the grid. CONCLUSIONS: The results of this series demonstrate that MSTs can be performed with acceptable morbidity in children undergoing epilepsy surgery. The precise role of MSTs in controlling seizure frequency and outcome, especially when combined with planned cortical resections, awaits further study. PMID- 14758940 TI - Cerebral hemispherectomy in pediatric patients with epilepsy: comparison of three techniques by pathological substrate in 115 patients. AB - OBJECT: Cerebral hemispherectomy for intractable seizures has evolved over the past 50 years, and current operations focus less on brain resection and more on disconnection. In addition, cases involving cortical dysplasia and Rasmussen encephalitis are being identified and surgically treated in younger individuals. Few studies have been conducted to compare whether there are perioperative differences based on hemispherectomy technique and/or pathological substrate in pediatric patients with epilepsy. METHODS: In this study the authors compared, stratified by disease, anatomical (37 cases) and Rasmussen functional hemispherectomy (32 cases) with a new modified lateral hemispherotomy (46 cases). Pathological processes included cortical dysplasia (55 cases), Rasmussen encephalitis (21 cases), infarction/ischemia (27 cases), and other/miscellaneous (12 cases). The authors found differences in perioperative clinical factors based on operative technique and/or pathological substrate. In terms of technique, the lateral hemispherotomy was associated with the least intraoperative blood loss, shortest intensive care unit stay, and lowest complication rate. The anatomical hemispherectomy was associated with the longest hospital stay, delayed oral food intake, highest postsurgery fevers, and the highest incidence of shunt requirement. The functional hemispherectomy was associated with the highest reoperation rate for recurrent seizures (25%). In terms of pathology, patients with cortical dysplasia were the youngest at surgery, suffered the greatest amount of blood loss, and required the longest operative/anesthesia times compared with the other pathologically defined groups. Postoperative seizure control (range 0.5-2 years) was not statistically different according to technique or disease process and was similar to that in cases of pediatric temporal lobe epilepsy. CONCLUSIONS: The authors found differences in perioperative risks and hospital course but not postsurgery seizure control, which vary by hemispherectomy technique and/or disease process. The modified lateral hemispherotomy approach offers various advantages related to operative blood loss and reoperation compared with anatomical and functional hemispherectomies that are especially relevant in younger patients with cortical dysplasia and Rasmussen encephalitis with small and/or malformed ventricles. PMID- 14758941 TI - Long-term outcome in children with moyamoya syndrome after cranial revascularization by pial synangiosis. AB - OBJECT: Moyamoya syndrome, a narrowing of the basal intracranial vessels accompanied by the development of a cloud of collateral "moyamoya" vasculature, causes cerebral ischemia and stroke. This study was undertaken to determine if a standardized neurosurgical revascularization procedure, pial synangiosis, conferred long-term benefit in pediatric patients. METHODS: The authors reviewed the clinical and radiographic records obtained in a consecutive series of patients with moyamoya syndrome. Patients were 21 years of age or younger and underwent surgery performed by a single neurosurgeon during a 17-year period. There were 143 patients (89 females and 54 males). Sixteen patients were Asian. Neurofibromatosis was present in 16 patients, 13 had undergone therapeutic cranial irradiation, and Down syndrome was present in 10. In 66 there was no known predisposing condition. Stroke had occurred in 67.8% of the population and transient ischemic attacks (TIAs) in 43.4% prior to surgery. Within the first 30 days following 271 craniotomies for pial synangiosis, there were 11 episodes of stroke (7.7% per patient; 4% per surgically treated hemisphere) and three severe TIAs. Follow-up evaluation was performed in all but one patient (mean period 5.1 years). In 126 patients followed for more than 1 year, four suffered a late-onset stroke, one suffered a severe reversible TIA without magnetic resonance imaging documented evidence of stroke, and two experienced persistent TIAs. In 46 patients followed for more than 5 years in whom the major initial presentation was stroke alone, only two late-onset strokes have occurred. Functional status at the time of surgery determined long-term functional status. CONCLUSIONS: Following pial synangiosis, the majority of pediatric patients with moyamoya syndrome stop having strokes and TIAs, and they appear to experience an excellent long-term prognosis. PMID- 14758942 TI - Use of gadolinium as an intraarterial contrast agent for pediatric neuroendovascular procedures. AB - OBJECT: The safety and efficacy of Gd as an intraarterial contrast agent for pediatric neuroendovascular procedures were investigated. METHODS: The authors retrospectively reviewed data regarding pediatric neuroendovascular procedures performed during a 1-year period. Seventeen procedures involving the use of gadodiamide in nine pediatric patients were identified. All angiographic images were of diagnostic quality but usually required minimal postimage processing. In time-consuming cases, the use of gadodiamide allowed the procedure to continue when the dosage of iodinated contrast approached a toxicity level and would have otherwise prompted termination of the procedure. This likely reduced the total number of procedures necessary. No adverse events due to the intraarterial use of gadodiamide were identified. CONCLUSIONS: Gadodiamide appears to be a safe and effective contrast agent for pediatric patients. PMID- 14758943 TI - Long-term effects of indirect bypass surgery on collateral vessel formation in pediatric moyamoya disease. AB - OBJECT: The authors investigated whether revascularization surgeries in children with moyamoya disease could reduce abnormal collateral formations in the posterior circulation; long-term follow-up angiography was performed to assess status. METHODS: In six patients who underwent indirect bypass surgery in childhood, long-term follow-up angiography studies were obtained between 7 and 12 years after surgery. All six patients presented with ischemic symptoms at onset of disease between 2 and 7 years of age. Ischemic insults did not occur after surgery in any patient. In five cases, the angiographically documented stages advanced bilaterally, and in three cases the angiography revealed nearly occluded intracranial carotid arteries bilaterally. In all sides in these five cases, the collateral circulation through the grafts developed well, and collateral formation from the lenticulostriate arteries and from the anterior choroidal artery decreased according to the natural advancement of disease. There was no reduction in the collateral formations from the posterior circulation, however, in four of these five cases. CONCLUSIONS: Although the data cannot directly indicate that the patients in this study remain at potential risk of developing hemorrhage in the future, these facts should be considered when evaluating the efficacy of revascularization surgeries, because the hemorrhagic potential of abnormally dilated collateral vessels from the posterior circulation would increase as the advancement of the disease. PMID- 14758944 TI - Failure of autologous bone-assisted cranioplasty following decompressive craniectomy in children and adolescents. AB - OBJECT: The authors have routinely performed primary autologous cranioplasty to repair skull defects after decompressive craniectomy. The high rates of subsequent bone resorption occurring in children prompted this study. METHODS: In an institutional review, the authors identified 40 (32 male and eight female) children and adolescents ranging from 4 months to 19 years of age in whom autologous cranioplasty was performed after decompressive craniectomy. The defect surface area ranged from 14 to 147 cm2. In all cases, the bone was fresh frozen at the time of the decompression. Symptomatic bone resorption subsequently occurred in 20 children (50%) in all of whom reoperation was required. The incidence of bone resorption significantly correlated with an increased skull defect area (p < 0.025). No significant correlation was found with age, sex, or anatomical location of the skull defect, number of fractured bone fragments, presence of a shunt, cause for decompressive craniectomy, method of duraplasty, or interval between the craniectomy and the cranioplasty. Reoperation to repair the resorbed autologous bone was performed 2 to 76 months after the initial procedure. CONCLUSIONS: The use of autologous bone to reconstruct skull defects in pediatric patients after decompressive craniectomy is associated with a high incidence of bone resorption. The use of autologous bone should be reevaluated in light of the high rate of reoperation in this pediatric population. PMID- 14758945 TI - Tumors of the skull base in children and adolescents. AB - OBJECT: Published data obtained in children with tumors of the skull base are sparse. In the majority of the available reports, the authors focus on the technical application of skull base approaches, but they contribute a paucity of information on the management of specific tumors, especially malignant skull base lesions. The purposes of this report are to increase the collective experience with the treatment of these tumors and to identify successful management paradigms. METHODS: The authors retrospectively reviewed the clinical records, pathological reports, and diagnostic images obtained in 24 children (< or = 19 years of age) with tumors arising from the cranial base in whom resection was part of their management between 1992 and 2002. Surgery-related complications and outcomes were analyzed with regard to tumor type and surgical approach. The median age of the group was 14 years. Tumors involved the anterior skull base in eight (33%), the middle skull base in 10 (42%), both the anterior and middle skull base in four (17%), and the posterior skull base in two patients (8%). Benign lesions were discovered in 11 patients (46%) and malignant neoplasms in 13 (54%). The tumors were most commonly of mesenchymal origin (21 [87.5%] of 24 tumors). Thirty surgical procedures were performed using a number of skull base approaches. A gross-total resection was achieved in 23 procedures (77%) and a subtotal resection in five (17%); a biopsy procedure was performed in one case; and the disease process could not be accessed in one case. One patient died in the perioperative period. Minor complications (Karnofsky Performance Scale score > or = 90, no prolongation of hospital stay, and no further surgery needed) occurred following 10 (33%) of the 30 surgical procedures. These affected 10 (42%) of the 24 patients and resulted in persistent or prolonged deficits in only five patients (21%). In nine patients (38%) the tumor recurred after a mean duration of 23 months. CONCLUSIONS: Skull base tumors in children affect mainly the anterior and middle cranial fossa. Sarcomas account for the majority of malignant tumors. Treatment of skull base tumors in children and adolescents needs to be tailored to patient age, tumor location, and tumor type. PMID- 14758946 TI - Long-term follow up of children with meningiomas in Denmark: 1935 to 1984. AB - OBJECT: Meningiomas in children are rare, infrequently described in the literature, and often associated with neurofibromatosis Type 2 (NF2). The authors report a series of 22 children treated for an intracranial meningioma in Denmark between 1935 and 1984. METHODS: Of 1542 cases of pediatric intracranial tumors in children younger than 15 years of age, 22 harbored meningiomas. Three children suffered from NF. The male/female ratio was 8:14. The mean age at the time of diagnosis was 5 years for boys and 11.5 years for girls. At the time of diagnosis all tumors were large. All patients underwent surgery. In 20 cases, the final histological diagnoses were low-grade and in two cases high-grade tumors. The follow-up period ranged from I to 45 years (mean 16 years). Two patients were lost to follow up. Four of seven boys and three of 13 girls survived. Five of 13 children in whom the tumor was completely removed survived, whereas two of seven in whom the lesion was partially removed survived. The mean survival time in children who died during follow up was 10 years. Two children with anaplastic meningioma remain alive. CONCLUSIONS: The long-term prognosis for surgically treated children with intracranial meningiomas was worse than expected. Some reasons for this may have been the late diagnosis and related large size of the tumor during a period of limited diagnostic capacity prior to the computerized tomography and magnetic resonance imaging eras, and the association of NF2 with multiple tumors of the central nervous system. Complete resection is not always possible and should be performed as an image-guided operation. PMID- 14758947 TI - Efficacy of scheduled nonnarcotic analgesic medications in children after suboccipital craniectomy. AB - OBJECT: The authors performed a study to evaluate the efficacy of a regimen of scheduled minor analgesic medications in managing postoperative pain in children undergoing intracranial procedures. METHODS: Postoperative pain scores were analyzed in two groups of children who underwent decompressive surgery for Chiari malformation: Group A underwent a scheduled regimen of minor oral analgesic medications (standing doses of acetaminophen [10 mg/kg] and ibuprofen [10 mg/kg] alternating every 2 hours) and Group B received analgesic medication when requested. Fifty children underwent a standard occipital craniectomy (25 in each group). The pain scores were significantly lower in Group A during most of the postoperative period. Length of stay (LOS) was shorter (2.2 compared with 2.8 days), and narcotic and antiemetic requirements were also lower in Group A patients than in Group B patients. Patients with spinal cord syringes exhibited a similar postoperative status to those without, and similar improvements in pain scores with scheduled minor analgesic medications were also evident. CONCLUSIONS: A regimen of minor analgesic therapy, given in alternating doses every 2 hours immediately after craniotomy and throughout hospitalization, significantly reduced postoperative pain scores and LOS in children in whom suboccipital craniotomy was performed. Narcotic and antiemetic requirements were also decreased in association with this regimen. Application of this postoperative analgesia protocol may benefit children and adults in whom various similar neurosurgical procedures are required. PMID- 14758949 TI - Cessation of chronic hypertension after posterior fossa decompression in a child with Chiari I malformation. Case report. AB - The authors report on the case of a patient evaluated for Valsalva maneuver induced headache, dizziness, and ataxia. Neuroimaging revealed a Chiari I malformation without syringomyelia. A history of idiopathic hypertension was noted. After posterior fossa decompression, pathologically elevated blood pressure was absent, and at 24-month follow-up evaluation the patient remained normotensive. Although seemingly rare, this case illustrates that some patients with tonsillar ectopia may exhibit elevated blood pressure. Clinicians should consider hindbrain herniation a rare cause in idiopathic hypertension. PMID- 14758948 TI - Detection of oncogene amplifications in medulloblastomas by comparative genomic hybridization and array-based comparative genomic hybridization. AB - OBJECT: Few studies have been conducted to investigate the genomic survey of oncogene amplification in medulloblastoma. Low frequency of N-myc, C-myc, and epidermal grow factor receptor (EGFR) gene amplification (< 10%) has been reported in medulloblastoma. Previous comparative genomic hybridization (CGH) study of primary medulloblastomas has revealed chromosomal amplification on 2p21, 3p, 5p15.3, 7q, 8q24, 11q22.3, and 17q. The aim of this study was to detect common oncogenes involved in medulloblastoma tumorigenesis. METHODS: The authors studied a series of 14 samples by performing CGH and array-based CGH. The CGH analysis detected nonrandom losses on 8p, 17p, 16q, 8q, and 1p, whereas gains were found on 17q, 12q, 7q, and 1p. Array-based CGH was conducted to investigate amplification of 58 oncogenes throughout the genome of these samples. Gene amplifications identified for the first time included PGY1 at 7q21.1, MDM2 at 12q14.3-q15, and ERBB2 at 17q21.2. The highest frequencies of oncogene gain were detected in D17S1670 (61.5%), PIK3CA (46.2%), PGY1 (38.5%), MET (38.5%), ERBB2 (38.5%), and CSE1L (38.5%). The gain in gene copy numbers was confirmed in 34 additional archival medulloblastoma cases by using fluorescence in situ hybridization analysis. CONCLUSIONS: This is the first genome-wide survey of multiple oncogene amplifications involved in the development of medulloblastoma. Gains of several candidate oncogenes such as D17S1670, ERBB2, PIK3CA, PGY1, MET, and CSE1L were frequently detected. These genes may be used as molecular markers and therapeutic targets of medulloblastomas. PMID- 14758950 TI - Extracorporeal membrane oxygenation combined with endovascular embolization for management of neonatal high-output cardiac failure secondary to intracranial arteriovenous fistula. Case report. AB - The authors describe a novel approach to the management of high-output heart failure secondary to an intracranial high-flow dural arteriovenous fistula (DAVF) by using extracorporeal membrane oxygenation (ECMO). To the best of the authors' knowledge, this represents the first report of an embolization performed in conjunction with the use of an ECMO circuit and the first report in which an ECMO cannula was used for intraarterial access for cerebral angiography. A 2-day-old girl presented with severe, high-output heart failure secondary to a high-flow intracranial DAVF. The patient was neurologically intact and no brain parenchymal abnormalities were revealed on computerized tomography scanning of the head, but she suffered severe heart failure, pulmonary hypertension, and liver and renal dysfunction. The patient underwent three endovascular embolization procedures involving coils and liquid embolic agents. Despite a decrease in the DAVF flow, the patient had only transient improvement in her pulmonary hypertension, and venoarterial ECMO therapy was instituted. Another embolization was performed while the patient was receiving ECMO therapy. Her cardiovascular status improved, she was weaned from ECMO, and she was eventually discharged home to her family. Extracorporeal membrane oxygenation can be used to sustain severely ill neonates with high-output heart failure secondary to intracranial AVFs. Embolization can be performed while the patient is receiving ECMO therapy. PMID- 14758951 TI - Intracranial pial arteriovenous fistulas with single-vein drainage. Report of three cases and review of the literature. AB - Intracranial pial arteriovenous fistulas (AVFs) are rare vascular lesions, of which fewer than 90 cases have been reported in the literature. They are composed of one or more arterial feeding vessels and a single draining vein that usually result in a huge varix. Because of the high-flow shunting, a poor prognosis is associated with conservative treatment. Surgical or endovascular treatment also poses various challenges. The authors present the cases of three patients in whom N-butyl-2-cyanoacrylate-assisted embolization was performed. Outcome in all three cases was good. The necessity of staged procedures to obliterate the AVF is also discussed. PMID- 14758952 TI - Migration of distal ventriculoperitoneal shunt catheter into the heart. Case report and review of the literature. AB - Migration of distal ventriculoperitoneal shunt tubing is known to occur in a wide of variety of locations. The authors report an unusual complication involving a previously confirmed intraperitoneal shunt catheter that migrated into the heart and pulmonary vasculature. Radiographic evidence suggested that this occurred secondary to cannulation of a segment of the external jugular vein with a shunt trochar during tunneling of the distal catheter. This is the sixth reported case of a peritoneal shunt tube migrating proximally into the heart. The authors review the literature regarding migration of distal tubing into the heart and pulmonary artery. Based on imaging studies obtained in the present case, the authors posit that the mechanism for this unusual type of shunt migration is inadvertent penetration of either the internal or external jugular vein during the initial tunneling procedure. Negative intrathoracic pressure and slow venous flow then draws the catheter out of the peritoneum and into the vasculature. The distal catheter then migrates into the right side of the heart and pulmonary artery. Diagnosis and management of this type of complication is discussed. PMID- 14758953 TI - Hypothalamic hamartoma secreting corticotropin-releasing hormone. Case report. AB - The authors describe a unique case of a 2-year-old boy with a hypothalamic hamartoma secreting corticotropin-releasing hormone (CRH). The patient presented with a history of behavioral disturbances progressing over 12 months. His neurological status was intact. Magnetic resonance imaging demonstrated a 1.8 x 1.6 x 1.2-cm isointense, nonenhancing hypothalamic lesion. Endocrinological workup revealed elevated serum CRH and adrenocorticotropic hormone levels, nonsuppression with low-dose dexamethasone, and partial suppression with high dose dexamethasone. He underwent tumor resection via a right frontotemporal craniotomy. Pathological examination of the tissue confirmed a hypothalamic hamartoma with CRH immunostaining. Postoperatively, his hormone levels normalized and his behavioral disturbances abated. The radiographic and clinical characteristics of hypothalamic hamartomas are reviewed and therapeutic considerations discussed. PMID- 14758954 TI - Hypothalamic ganglioglioma treated by temporal lobectomy. Case report and review of the literature. AB - The authors report the case of a hypothalamic ganglioglioma with left-sided temporal lobe extension in an 8-year-old girl who presented with seizures. Other cases of ganglioglioma involving the hypothalamus have been reported in the literature; however, this site of origin is exceedingly rare and worthy of report. Treatment involved medial temporal lobectomy with the hypothalamic component of the tumor remaining untouched. The patient recovered postoperatively with no neurological deficits and was seizure free at 12 months. Neither radio- nor chemotherapy was recommended because of the tumor histology, location, and the patient's age. The authors recommend follow up and surgical treatment for possible tumor recurrence. The prognosis for hypothalamic ganglioglioma is unknown. PMID- 14758955 TI - Nonsurgical treatment of a C6-7 unilateral locked facet joint in an infant. Case report. AB - Cervical spine injuries in the pediatric population typically affect the upper cervical region. The authors present the first reported case of a subaxial C6-7 unilateral locked facet joint in a neurologically intact 10-month-old infant. To date, this patient's nonoperative treatment has been successful. The proposed biomechanical mechanism of this injury and the treatment paradigm are discussed. PMID- 14758956 TI - Cervical cord exostosis compressing the axis in a boy with hereditary multiple exostoses. Case illustration. PMID- 14758957 TI - Choroid plexus papilloma of the third ventricle in the fetus. Case illustration. PMID- 14758958 TI - Inauguration of pediatric neurosurgery by Harvey W. Cushing: his contributions to the surgery of posterior fossa tumors in children. Historical vignette. AB - Development of posterior fossa surgery remains Harvey Cushing's hallmark contribution to pediatric neurosurgery. During the era before Cushing, posterior fossa lesions were considered inoperable, and only osseous decompressive surgery was offered. The evolution of Cushing's surgical expertise from subtemporal decompressions to total extirpation of vascular fourth ventricular tumors, combined with a dramatic decrease in his operative mortality rate, reflects the maturation of modern neurosurgical techniques. A comprehensive review of the medical records of Cushing's pediatric patients treated between 1912 and 1932 revealed that procedures such as lateral ventricular puncture (to decrease cerebellar herniation), transvermian approach to midline tumors, and electrocoagulation were the key factors punctuating the path to his pioneering achievements in posterior fossa surgery. The outcome of such operations was improved by his recognition of the importance of tumor mural nodule in cyst recurrence, as well as elucidation of the histogenesis of pediatric posterior fossa tumors to tailor treatment including radiotherapy. PMID- 14758959 TI - Peritoneal cerebrospinal fluid shunt insertion. PMID- 14758960 TI - Applying a 'stages of change' model to enhance a traditional evaluation of a research transfer course. AB - The aim of this study was to utilize an evaluation tool based on Prochaska's model of change in order to assess behaviour change as part of an evaluation process for a research transfer training programme (RTTP). The RTTP was a training programme offered to scientists in a psychiatry department and research institute to gain skills in research transfer. In addition to a traditional course evaluation framework evaluating overall satisfaction with the course and whether or not learning objectives were met, an additional 'stages of change' evaluation tool designed to assess change along a continuum was utilized. This instrument measured change in participants' attitudes, intentions and actions with respect to research transfer practice and consisted of a 12-question survey completed by participants prior to taking the course and 3 months post-course. In two out of the three categories, attitudes and intention to practice, there was positive change from pre- to post-course (P < 0.05). Although there was a trend of increased RT-related action, this was less robust and did not reach significance. For the RTTP transfer course, a 'stages of change' model of evaluation provided an enhanced understanding by showing changes in participants that would otherwise have been overlooked if only changes in RT behaviour were measured. Additionally, evaluating along a change continuum specifically identifies areas for improvement in future courses. The instrument developed for this study could also be used as a pre-course, participant needs assessment to tailor a course to the change needs of participants. Finally, this 'stages of change' approach provides insight into where barriers to change may exist for research transfer action. PMID- 14758961 TI - DARTS 2000 online diabetes management system: formative evaluation in clinical practice. AB - RATIONALE, AIMS AND OBJECTIVES: Failure to engage in user-informed evaluation of emergent health informatics tools can have negative consequences for future implementation, related both to poor usability or clinical utility and to suboptimal stakeholder buy-in. This paper describes a formative evaluation in primary care of a multifaceted, web-based resource for diabetes management. The primary aims were to assess the usability and utility of the prototype in order to inform system refinements prior to implementation, and to investigate barriers and facilitators to system use so as to aid the development of a tailored implementation plan. METHODS: A mixed-method approach involving survey, remote observation, semi-structured interviews and electronic feedback. CONTEXT: One Scottish local health care cooperative comprising five general practice surgeries and their staff. RESULTS: A survey following temporary exposure to a dummy site revealed high levels of computer familiarity, welcoming attitudes and positive ratings of usability, format and utility. Comments mainly addressed content accuracy, feature suggestions and usability issues. Key barriers and facilitators to use included time and training. Remote observation following access to live clinical data enabled profiling of usage by individual and professional group. In the 3 month observation period administrators were the most frequent users, followed by GPs and nurses, with clinical and data entry screens accessed most often. Semi-structured interviews with key respondents sampled by professional group and usage frequency provided richer qualitative information on barriers and facilitators to use, patterns of system integration into work routines and system usability, content and utility. Content analysis of electronic feedback revealed mainly technical queries and general expressions of satisfaction. CONCLUSIONS: Evaluation informed a number of important and unforeseen improvements to the prototype and helped refine the implementation plan. Engagement in the process of evaluation has led to high levels of stakeholder ownership and widespread implementation. PMID- 14758962 TI - Methodology for evaluating physician order entry (POE) implementations. AB - The body of physician order entry (POE) implementations literature uses statistical evaluation methods to demonstrate changes in specified variables after POE implementation. To understand and manage the holistic impact of POE on the health care institution, a methodology that utilizes feedback to guide the POE implementation towards the satisfaction of stakeholder objectives is presented. Stakeholders jointly define quantitative and qualitative metrics for their objectives, establish target value vectors for the metrics that represent acceptable implementation outcomes and specify evaluation milestones. These are used to compare pre- and post-POE implementation clinical performance, enabling a socio-technical feedback-improvement cycle. A case study is provided to illustrate how the methodology is being used at Sunnybrook and Women's College Health Science Centre in Toronto, Canada. PMID- 14758963 TI - Physician gender and antihypertensive prescription pattern in primary care. AB - OBJECTIVES: To determine: (i) the gender-based differences of physicians in prescribing antihypertensive drugs in the management of hypertension; (ii) the influence of patient comorbidity such as diabetes mellitus on the gender-based pattern of antihypertensive prescription, and (iii) gender-based prescription patterns among family physicians (FPs) and general practitioners (GPs). METHODS: A survey study was carried out at 15 out of 20 health centres in Bahrain during May and June 2000. A total of 3971 prescriptions, issued to 2705 patients with uncomplicated hypertension and 1266 patients with diabetic hypertension by 77 FPs (female = 54, male = 23) and 41 GPs (female = 11, male = 30), were analysed. RESULTS: As monotherapy, female physicians preferred ACE inhibitors (OR: 0.82, CI: 0.68-0.98, P = 0.033). In terms of overall drug utilization (monotherapy + combination therapy): (i) male physicians preferred beta blockers (OR: 1.17, CI: 1.03-1.31, P = 0.014) and diuretics (OR: 1.15, CI: 1.00-1.32, P = 0.047), while female physicians preferred methyldopa (OR: 0.73, CI: 0.56-0.94, P = 0.019); (ii) in uncomplicated hypertension, female physicians preferred calcium channel blockers (OR: 0.83, CI: 0.69-0.99, P = 0.038) and methyldopa (OR: 0.69, CI: 0.49 0.98, P = 0.042), and (iii) in diabetic hypertension, male physicians preferred beta blockers (OR: 1.26, CI: 1.00-1.57, P = 0.047). While female FPs prescribed methyldopa more extensively (OR: 0.66, CI: 0.47-0.92, P = 0.018), male GPs prescribed beta blockers (OR: 1.28, CI: 1.00-1.62, P = 0.046). CONCLUSIONS: Within the same practice setting, gender-based differences in the prescription of antihypertensive drugs were seen. Such preference for a particular class of antihypertensives was also influenced by the presence of comorbidity in patients and, to a limited extent, by the training level of primary care physicians. PMID- 14758964 TI - The influence of medical professionalism on scientific practice. AB - This paper examines how the practise of science in medicine may be subverted by the professionalism of medicine. The requirements of science as regards the axiom of the burden of proof and research design may be inevitably entangled with the influence of authority. This entanglement may be particularly strong in medical research because of the way the profession works and is organized. The nature and possible effects of this entanglement are explored by examining the cultural differences between scientists and professionals and their approach to authority. Then the nature of the axiom of burdens of proof and research design are described, followed by accompanying discussions of how aspects of authority and professional organization may influence them. Finally, concluding comments address ways forward. PMID- 14758965 TI - A qualitative evaluation of information leaflets for gastroscopy procedure. AB - Written information sent to patients prior to diagnostic gastroscopy is an important part of the process of informing and preparing them for the procedure. Yet there is ample evidence in the literature that information leaflets do not measure up to the required standard. In this study, information leaflets from a random sample of seven hospitals in Northern Ireland that carried out gastroscopy as a day procedure were evaluated using a checklist of items recommended by the British Society of Gastroenterology (BSG) for inclusion in leaflets for patients undergoing diagnostic gastroscopy. The results showed that the number of written materials sent to patients prior to the procedure varied between units. There were inconsistencies in the information given by the same unit, and overall, there was a lack of vital information in most of the leaflets. Some of the information was confusing and ambiguous. The potential risk of the procedure was explained in only one of the leaflets. Patients' right to choose to have a mild sedative was not made clear in most of the leaflets. More should be done to address these gaps and inconsistencies in the written information provided to patients prior to gastroscopy. PMID- 14758966 TI - Improving blood gas control in mechanically ventilated, premature infants through monitoring and evaluation of clinical practice. AB - Mechanical ventilation causing hypocapnia or hyperoxia carries a risk for the pre term infant. The aim was to improve blood gas control in our unit. A guideline was written, and all personnel were motivated concerning blood gas control. Case records of all mechanically ventilated premature infants were examined during two 3-month periods, before and after intervention. The hours spent with hypocapnia (pCO2 < 4 kPa) or hyperoxia (PO2 > 12 kPa) were recorded. Case records of 31 infants were examined for a total of 1358 h of mechanical ventilation, 641 h before and 717 h after the intervention. The percentage time of hypocapnia before intervention (7.0%) was reduced significantly (P = 0.044) to less than half (2.9%) after intervention. Hyperoxia was reduced from 14.5% to 8.7% (P = 0.072). Blood gas control of mechanically ventilated premature infants could be improved with little effort, but hyperoxia is too frequent. PMID- 14758967 TI - Moral activism and the nursing profession: meeting the challenge to be involved. PMID- 14758968 TI - New ICN policy promotes nurses, nursing and health. PMID- 14758969 TI - ICN mobile libraries reach refugee camps in Tanzania. PMID- 14758970 TI - Iraqi nurses develop a national strategic plan. PMID- 14758971 TI - Patient safety concerns us all. PMID- 14758972 TI - Childhood smoking: the research, clinical and theoretical imperative for nursing action. AB - BACKGROUND: Tobacco smoking is a major international health issue which nurses are ideally positioned to address. Childhood represents a critical period for intervention to prevent/reduce smoking. During childhood, the majority of smokers first experiment with smoking, are initiated into the smoking subculture and become addicted to tobacco. Children are highly susceptible to smoking as a result of developmental factors, which promote or facilitate high-risk behaviours, limited coping skills, limited defences and inadequate legal protection against youth-focused marketing of tobacco. AIM: This paper is intended to sensitize nurses to the magnitude of childhood smoking as an international health problem and to familiarize them with current intervention approaches and care issues pertinent to child and adolescent populations. METHOD: An extensive literature review was conducted to determine the impact of childhood smoking at the personal and community level, characteristics of child smokers, the processes associated with smoking cessation, intervention approaches and intervention models for use by nurses across the spectrum of clinical settings. FINDINGS: Evidence exists that behaviourally based interventions by nurses for smoking prevention/cessation are effective with children. Key components of effective behaviourally based antismoking approaches for smokers include addressing self-efficacy to quit, providing social support, resisting temptation to smoke and discussing issues related to relapse and relapse prevention. For all children, the reinforcement of non-smoking behaviour is essential. Intervention models, which can be incorporated into routine clinical care of individual children or with groups, emphasize the importance of parental involvement, routine screening for tobacco use and provision of a clear message that smoking and tobacco use is unhealthy. PMID- 14758973 TI - Women and the global tobacco epidemic: nurses call to action. AB - BACKGROUND: Tobacco consumption is increasing among women across the globe at alarming rates. Without effective intervention, the smoking prevalence among women will nearly triple over the next generation. These trends are potentially more threatening when considering how tobacco intersects women's lives, regardless if they use tobacco products or not. AIM: A review and analysis of the literature is conducted to examine the scope of tobacco's global effect on the multiple dimensions of women's health. METHOD: Medline (1990-2003), Cumulative Index to Nursing and Allied Health Literature (1990-2003) and World Health Organization databases were searched for related topics. Keywords for searches included global health, tobacco, women and nursing. FINDINGS: The epidemiology and prevalence of tobacco use among women are presented and its impact on women globally. Using an ecological perspective, the consequences of tobacco are analysed within the contexts of health, social, environment, economic and policy as it relates to women, their families and their communities. IMPLICATIONS: Nurses are in prime positions to empower individuals, families, communities and nations in the prevention and treatment of tobacco use. Health for all women continues to be a call for equity and social justice. Recommendations are provided for nursing practice, education, theory, research and policy to address this global health concern. PMID- 14758974 TI - Adolescent mothers' utilization of contraceptive services in South Africa. AB - BACKGROUND: In South Africa, contraceptives, as well as emergency contraceptives, are available free of charge. Since 1996, changed legislation has enabled women of all ages to choose whether they wish their pregnancies to be terminated during the first 12 weeks of pregnancy. Therefore, adolescent mothers, 19 years or younger at the time of their babies' birth, were investigated regarding why they failed to use contraceptive, emergency contraceptive or termination of pregnancy services. AIM: To explore the knowledge of young mothers regarding contraception. METHOD: An exploratory descriptive survey, utilizing questionnaires and convenience sampling. FINDINGS: The majority of the participating 250 adolescent mothers lacked knowledge about contraceptives, emergency contraceptives and termination of pregnancy services. Merely legalizing the termination of pregnancies, and providing free contraceptive and emergency contraceptive services, did not affect the utilization of these services by the 250 adolescent mothers investigated. CONCLUSION: Young mothers require more knowledge to enable them to make better informed decisions, and the services need to become more readily accessible and user friendly to adolescents. Reproductive health services provided specifically to adolescents could enhance the utilization of such services. PMID- 14758975 TI - HIV infection of women in African countries. AB - BACKGROUND: The increase in human immunodeficiency virus infection among women of childbearing age increases the potential risk for vertical transmission of infection to their newborns. AIM: This paper discusses literature reviewed on human immunodeficiency virus infection of women in Africa with reference to Ghana, West Africa. The paper provides an epidemiological overview of human immunodeficiency virus infection of women in Ghana and Africa and the potential risk for vertical transmission and associated contributing factors. Literature reviewed was retrieved from the Cumulative Index of Nursing and Allied Health Literature and United States National Library of Medicine databases. CONCLUSION AND IMPLICATIONS FOR PRACTICE: Findings from the review of literature suggest that human immunodeficiency virus infection of African women is a major public health problem which must be addressed and considered a priority in the next decade. Leadership in African countries including nurses and policy-makers will have to do more to make a difference to the epidemic. Measures to reduce transmission of human immunodeficiency virus are discussed. PMID- 14758976 TI - Web-based cheminformatics and molecular property prediction tools supporting drug design and development at Novartis. AB - Web-based tools offer many advantages for processing chemical information, most notably ease of use and high interactivity. Therefore more and more pharmaceutical companies are using web technology to deliver sophisticated molecular processing tools directly to the desks of their chemists, to assist them in the process of designing and developing new drugs. In this paper, the web based cheminformatics system developed at Novartis and currently used by more than thousand users is described. The system allows various molecular modeling and molecular processing tasks, including the calculation of molecular and substituent properties, property-based virtual screening, visualization of molecules, bioisosteric design, diversity analysis, and support of combinatorial chemistry. The methodology to calculate various molecular properties relevant to drug design is described, including the prediction of intestinal absorption, blood-brain barrier penetration, efflux, and water solubility. Information about the web technology used is also provided. PMID- 14758977 TI - e-molecular shapes and properties. AB - Due to recent computer technology advances, shape analysis has gained importance in all domains. In drug design and proteomics, molecular surfaces (van der Waals surface, solvent accessible surface, solvent excluded surface, polar surface area, electron density surface, separating surface, etc.), buried surfaces (gap, cleft, cavity, etc.) as well as shape properties of these surfaces, can be easily computed and visualized via the Internet. Freely available resources from the Internet for academic use, are reviewed. PMID- 14758978 TI - Prediction of biological activity spectra via the Internet. AB - The majority of biologically active compounds have both pharmacotherapeutic and side/toxic actions. To estimate general efficacy and safety of the molecules under study, their biological potential should be thoroughly evaluated. In an early stage of study, only information about structural formulae was available and was used as an input for computational prediction. Based on a structural formulae of compounds presented as SDF or MOL-files, computer program PASS predicts 900 pharmacological effects, mechanism of action, and specific toxicity. An average accuracy of prediction in leave-one-out cross-validation is about 85%. For evaluating new compounds, scientific community may use PASS via the Internet for free at URL: http://www.ibmh.msk.su/PASS. In the first 18 months of PASS Inet's use, approximately 1000 researchers from 60 countries have obtained predicted biological activity spectra for about 23,000 different chemical compounds. More than 64 million PASS predictions for almost 250,000 compounds from Open NCI database are available on the web site http://cactus.nci.nih.gov/ncidb2/. These predictions are used for selecting compounds with desirable and without unwanted types of biological activities among the NCI samples available for screening. PMID- 14758979 TI - Zherlock: an open source data analysis software. AB - Zherlock is an open source software that provides state-of-the-art data analysis tools to the user in an intuitive and flexible way. It is a front-end to different numerical "engines" to produce a seamless integration of algorithms written in different computer languages. Of particular interest is creating an interface to high-level scientific languages such as Octave (a Matlab clone) and R (an S-PLUS clone) to enable efficient porting of new data analytical methods. Zherlock uses advanced scientific visualization tools in 2-D and 3-D and has been extended to work on virtual reality (VR) systems. Central to Zherlock is a visual programming environment (VPE) which enables diagram based programming. These diagrams consist of nodes and connection lines where each node is an operator or a method and lines describe the flow of data between nodes. A VPE is chosen for Zherlock because it forms an effective way to control the processing pipeline in complex data analyses. The VPE is similar in functionality to other programs such as IRIS Explorer, AVS or LabVIEW. PMID- 14758980 TI - In silico modelling of hazard endpoints: current problems and perspectives. AB - Major scientific hurdles in the acceptance of quantitative structure-activity relationships (QSAR) for regulatory purposes have been identified. First, when quantifying important features of chemical structure complexities of molecular structure have often been ignored. More mechanistic modelling of chemical structure should proceed on two fronts: by developing a more in-depth understanding and representation of the multiple states possible for a single chemical by achieving greater rigor in understanding of conformational flexibility of chemicals; and, by considering families of activated metabolites that are derived in biological systems from an initial chemical substrate. Second, QSAR research is severely limited by the lack of systematic databases for important risk assessment endpoints, and despite many decades of research, the ability to cluster reactive chemicals by common toxicity pathways is in its infancy. Finally, computational tools are lacking for defining where a specific QSAR is applicable within the domain (universe) of chemical structures that are to be regulated. This paper describes some of the approaches being taken to address these needs. Applications of some of these new approaches are demonstrated for the prediction of chemical mutagenicity, where considerations of both molecular flexibility and metabolic activation improved the QSAR predictability and interpretations. Lastly, the applicability domain for a specific QSAR predicting estrogen receptor binding is presented in the context of a mechanistically-defined chemical structure space for large heterogeneous chemical datasets of regulatory concern. A strategic approach is discussed to selecting chemicals for model improvement and validation until regulatory acceptance criteria for risk assessment applications are met. PMID- 14758981 TI - Comparative molecular field analysis (CoMFA) model using a large diverse set of natural, synthetic and environmental chemicals for binding to the androgen receptor. AB - A large number of natural, synthetic and environmental chemicals are capable of disrupting the endocrine systems of experimental animals, wildlife and humans. These so-called endocrine disrupting chemicals (EDCs), some mimic the functions of the endogenous androgens, have become a concern to the public health. Androgens play an important role in many physiological processes, including the development and maintenance of male sexual characteristics. A common mechanism for androgen to produce both normal and adverse effects is binding to the androgen receptor (AR). In this study, we used Comparative Molecular Field Analysis (CoMFA), a three-dimensional quantitative structure-activity relationship (3D-QSAR) technique, to examine AR-ligand binding affinities. A CoMFA model with r2 = 0.902 and q2 = 0.571 was developed using a large training data set containing 146 structurally diverse natural, synthetic, and environmental chemicals with a 10(6)-fold range of relative binding affinity (RBA). By comparing the binding characteristics derived from the CoMFA contour map with these observed in a human AR crystal structure, we found that the steric and electrostatic properties encoded in this training data set are necessary and sufficient to describe the RBA of AR ligands. Finally, the CoMFA model was challenged with an external test data set; the predicted results were close to the actual values with average difference of 0.637 logRBA. This study demonstrates the utility of this CoMFA model for real-world use in predicting the AR binding affinities of structurally diverse chemicals over a wide RBA range. PMID- 14758982 TI - Comparative toxicity and hazards of pesticides to Apis and non-Apis bees. A chemometrical study. AB - The adverse effects of 158 pesticides to the Honey bee (Apis mellifera), the alfalfa leafcutting bee (Megachile rotundata) and the alkali bee (Nomia melanderi) were compared by means of various linear and non-linear multivariate analyses. A comparison exercise including the bumble bee (Bombus spp.) was also performed from a more restricted set of 32 pesticides. While no difference of sensitivity was found between A. mellifera and Bombus spp., M. rotundata appeared the most susceptible to pesticides followed by N. melanderi. PMID- 14758983 TI - Prediction of substances of weak toxicity from chemical structures. An epidemiological approach. AB - A global structure set of acute toxicity (LD50, mouse, i.v.) was constructed from databases. The common structural features of low toxicity substances were obtained from 1495 structure-toxicity data by applying the substructural balance method (SBM). This set contained 111 low toxicity (> ca. 1000 mg/kg) and 817 high toxicity substances (< ca. 30 mg/kg). The result was expressed by a control chart which is made to control the structural causes to obtain the desired effect (low toxicity). Within the control limits, 100% of low toxic and 20% of high toxic substances were found. The control chart was tested with another set of 81 structures of low toxicity selected among 718 structures retrieved from RTECS. Seventy-eight structures were found within the control limits and the predictability was 96.3% (78/81). SBM employs a new paradigm of descriptors, the ratios of numbers of substructures, which were selected through a descriptor screening process. The results confirm that the new paradigm is applicable for structure-based toxicity prediction. This "epidemiological" approach suggests key structural criteria where no reliable mechanism of toxicity is available and serves to reduce the search space of organic structures. A theory supporting SBM was discussed. PMID- 14758984 TI - A novel method for GPCR recognition and family classification from sequence alone using signatures derived from profile hidden Markov models. AB - G-protein coupled receptors (GPCRs) constitute a broad class of cell-surface receptors, including several functionally distinct families, that play a key role in cellular signalling and regulation of basic physiological processes. GPCRs are the focus of a significant amount of current pharmaceutical research since they interact with more than 50% of prescription drugs, whereas they still comprise the best potential targets for drug design. Taking into account the excess of data derived by genome sequencing projects, the use of computational tools for automated characterization of novel GPCRs is imperative. Typical computational strategies for identifying and classifying GPCRs involve sequence similarity searches (e.g. BLAST) coupled with pattern database analysis (e.g. PROSITE, BLOCKS). The diagnostic method presented here is based on a probabilistic approach that exploits highly discriminative profile Hidden Markov Models, excised from low entropy regions of multiple sequence alignments, to derive potent family signatures. For a given query, a P-value is obtained, combining individual hits derived from the same family. Hence a best-guess family membership is depicted, allowing GPCRs' classification at a family level, solely using primary structure information. A web-based version of the application is freely available at URL: http:/bioinformatics.biol.uoa.gr/PRED-GPCR. PMID- 14758985 TI - New fuzzy logic strategies for bio-molecular recognition. AB - The concepts of molecular similarity and molecular complementarity, playing important roles in the broad field of molecular recognition, are chemical problems, in which the eyeball technique used by a human observer is very successful but which are very hard to code into a computer algorithm. Based on the model of molecular surfaces, our new approach defines overlapping surface patches with similar molecular properties. These patches are used to represent local features of the molecule in a way, which is beyond the atomistic resolution but can nevertheless be applied in partial similarity as well as complementarity analyses in a very general sense. It is shown that this molecular description can be used as the first step in a docking algorithm for complexes, where the structures of both molecules are known, as well as for the identification of possible active sites without the knowledge of specific molecules binding to this site. PMID- 14758986 TI - Comparison of MLR, PLS and GA-MLR in QSAR analysis. AB - The use of the internet has evolved in quantitative structure-activity relationship (QSAR) over the past decade with the development of web based activities like the availability of numerous public domain software tools for descriptor calculation and chemometric toolboxes. The importance of chemometrics in QSAR has accelerated in recent years for processing the enormous amount of information in form of predictive mathematical models for large datasets of molecules. With the availability of huge numbers of physicochemical and structural parameters, variable selection became crucial in deriving interpretable and predictive QSAR models. Among several approaches to address this problem, the principle component regression (PCR) and partial least squares (PLS) analyses provide highly predictive QSAR models but being more abstract, they are difficult to understand and interpret. Genetic algorithm (GA) is a stochastic method well suited to the problem of variable selection and to solve optimization problems. Consequently the hybrid approach (GA-MLR) combining GA with multiple linear regression (MLR) may be useful in derivation of highly predictive and interpretable QSAR models. In view of the above, a comparative study of stepwise-MLR, PLS and GA-MLR in deriving QSAR models for datasets of alpha1-adrenoreceptor antagonists and beta3-adrenoreceptor agonists has been carried out using the public domain software Dragon for computing descriptors and free Matlab codes for data modeling. PMID- 14758987 TI - QSAR studies on P-glycoprotein-regulated multidrug resistance and on its reversal by phenothiazines. AB - Multidrug resistance is brought about largely by membrane transport proteins such as P-glycoprotein (P-gp). We have developed a quantitative structure-activity relationship (QSAR) for P-gp-associated ATPase activity for a diverse set of 22 drugs, and found that such activity is related to substrate molecular size and polarity. We have also developed a QSAR for drug efflux from the blood-brain barrier of another diverse set of 22 drugs, and found that such efflux is a function of drug size and polarisability. Thirdly, we have carried out a QSAR analysis of the ability of 157 phenothiazines and related drugs to reverse multidrug resistance. We were unable to obtain a good QSAR for the whole data set, but when we divided the data-set into sub-sets of closely related structures, a series of good correlations was obtained, most of which incorporated descriptors that model molecular size and polarity/polarisability. In no instance did we find any evidence that hydrogen bonding or hydrophobicity play a part in multidrug resistance or its reversal, despite that fact that several other workers have reported that these effects appear to be important here. PMID- 14758988 TI - Comparative study of non nucleoside inhibitors with HIV-1 reverse transcriptase based on 3D-QSAR and docking. AB - The intermolecular interaction between two types of non nucleoside reverse transcriptase inhibitors (NNRTIs), HEPT and TIBO, and HIV reverse transcriptase receptor (HIVRT) was investigated. The result of docking study showed that two types of NNRTIs presented similar interaction mechanism with HIVRT. The most active compound of every type of inhibitors could form one hydrogen bond with the residue Lys101 and has hydrophobic interaction with residues Tyr181, Tyr188 and Tyr318, etc. Three 3D-QSAR models including two partial correlation models (one for each family of HEPT and TIBO) and a mixed model gathering two families were constructed. Comparative study of these models indicated that the mixed model offered the strongest prediction ability. For this model, the cross-validated q2 values were 0.720 and 0.675, non-cross-validated r2 values were 0.940 and 0.920 for CoMFA and CoMSIA, respectively. It has been validated by using a test set of 27 inhibitors. Compared with previously reported works, our model showed better prediction ability. It could help us to insight the interaction between NNRTIs and HIVRT, and to design new anti-HIV NNRTIs inhibitors. PMID- 14758989 TI - Theoretical comparison between structural and dynamical features of Dolastatins 11 and 12 antineoplastic depsipeptides. AB - Molecular mechanics (MM) and dynamics (MD) calculations in vacuo and in water have been performed for the natural cyclodepsipeptides Dolastatins 11 and 12 isolated from the sea hare Dolabella auricularia. The analysis of the MD trajectories for the two systems can give useful insight on the backbone structural features, side-chain and peptide-water interactions as well as on the inter- and intra-molecular hydrogen bonds. A comparison between the selected and analysed lowest energy isomers shows the different conformational behaviour of the compounds. Finally, with the aim to ascertain a structure-activity relationship for the two peptides, the interactions of both Dolastatins with water, generic hydrophobic environment, magnesium and calcium ions have been investigated by means of the GRID program. PMID- 14758991 TI - Reply to the comment on "Multiple scattering in a reflecting cavity: application to fish scattering". PMID- 14758992 TI - A simplified algorithm for second-order sound beams with arbitrary source distribution and geometry. PMID- 14758990 TI - Study of local anesthetic activity of some derivatives of 3-amino-benzo-[d] isothiazole. AB - On the basis of computer prediction of biological activity by PASS and toxicity by DEREK, the most prospective 18 alkylaminoacyl derivatives of 3-amino-benzo-[d] isothiazole were selected. Their local anesthetic action was assessed using an in vitro preparation of the isolated peroneal nerve of the frog. The local anesthetics action of the compounds was assessed according to the time required for each compound to reduce the amplitude of the evoked compound action potential (CAP). Lidocaine was used as the control compound. The results show that the tested compounds can be divided into three groups: (a) compounds with action similar to lidocaine, (b) compounds with action lower than lidocaine and (c) compounds which block completely the evoked CAP, but after the compound was removed and replaced with normal saline showed no recovery of the potential at all. QSAR studies showed that polarizability, polarity and presence of five membered rings in molecules have a positive influence on local anesthetic activity, while contributions of aromatic CH and singly bonded nitrogen are negative. Since estimations from PASS probabilities to find local anesthetic activity in the most active compounds were less than 50%, these compounds may be considered as new chemical entities (NCEs). PMID- 14758993 TI - Effect of talker and speaking style on the speech transmission index. PMID- 14758994 TI - The diffraction of sound by an impedance sphere in the vicinity of a ground surface. AB - The problem of sound diffraction by an absorbing sphere due to a monopole point source was investigated. The theoretical models were extended to consider the case of sound diffraction by an absorbing sphere with a locally reacting boundary or an extended reaction boundary placed above an outdoor ground surface of finite impedance. The separation of variables techniques and appropriate wave field expansions were used to derive the analytical solutions. By adopting an image method, the solutions could be formulated to account for the multiple scattering of sound between the sphere and its image near a flat acoustically hard or an impedance ground. The effect of ground on the reflected sound fields was incorporated in the theoretical model by employing an approximate analytical solution known as the Weyl-van der Pol formula. An approximation solution was suggested to determine the scattering coefficients from a set of linearly coupled complex equations for an absorbing sphere not too close to the ground. The approximate method substantially reduced the computational time for calculating the sound field. Preliminary measurements were conducted to characterize the acoustical properties of an absorbing sphere made of open cell polyurethane foam. Subsequent experiments were carried out to demonstrate the validity of the proposed theoretical models for various source/receiver configurations around the sphere above an acoustically hard ground and an impedance ground. Satisfactory comparative results were obtained between the theoretical predictions and experimental data. It was found that the theoretical predictions derived from the approximate solution agreed well with the results obtained by using the exact solutions. PMID- 14758995 TI - Measurement of six acoustical properties of a three-layered medium using resonant frequencies. AB - An ultrasonic technique for a simultaneous determination of traveling times and acoustic impedances of three-layered, platelike specimens using a normally incident longitudinal wave is presented. The acoustical properties are determined by minimizing the difference between measured and calculated resonant frequencies in the well-known least-squares sense. The resonant frequencies of the layered structure are deduced, in transmission, from the maxima of the magnitude of the transmission spectrum, or, in reflection, from the pi values of the phase of the reflection spectrum. Measurements are carried out in the frequency range 1-20 MHz for steel/aluminum/steel and silver/copper/nickel specimens with individual layer thickness values ranging from 0.3 mm to 2 mm. The differences between the inverted thicknesses and those measured directly with an optical microscope range from -4 microm to +13 microm. Estimated error bounds suggest that the inverted travel times are accurate within +/- 1 ns with 95% probability. PMID- 14758996 TI - Measurement of dependence of backscatter coefficient from cylinders on frequency and diameter using focused transducers--with applications in trabecular bone. AB - A theory for the elastic scattering response from a cylinder insonified by a plane wave was previously derived by Faran. In the present paper, the empirical relationship between Faran's theory and measurements of backscatter coefficient from cylindrical targets using focused transducers is investigated. Experimental measurements of dependence of backscatter coefficient on frequency and diameter for nylon wires are reported. It is found that, under certain conditions (including weak, incoherent scattering), backscatter coefficient measurements from collections of cylindrical scatterers may be meaningfully compared with Faran's model predictions. At low frequencies, the theory and experimental measurements exhibit similar dependences on frequency and diameter, provided that the scatterers are not too densely packed. At higher frequencies, the fine structure of Faran's predictions becomes difficult to reproduce experimentally with a focused transducer. Implications regarding applications to characterization of trabecular bone are discussed. PMID- 14758997 TI - The role of the coupling term in transient elastography. AB - The transient radiation of low-frequency elastic waves through isotropic and homogeneous soft media is investigated using the Green's function approach. A careful analysis of the coupling term is performed and yields the introduction of a very near field region in which its amplitude behaves as 1/r. To address the calculation of impulse responses, a simplified Green's function is proposed for semi-infinite media and compared to exact solutions. Impulse response calculations are successfully compared with experimental measurements obtained for circular radiators of different diameters using transient elastography. Results presented in this paper provide a better understanding of the role of the coupling term in elastography and should be used to compensate diffraction and coupling effects observed in transient elastography. PMID- 14758998 TI - Acoustic cavity modes in lens-shaped structures. AB - A method for solving exactly the Helmholtz equation in parabolic rotational coordinates is presented using separability of the eigenfunctions and the Frobenius power series expansion technique. Two examples of interest in acoustics are considered and analyzed quasianalytically: The acoustic pressure in a cavity defined by two paraboloids (forming a lens-shaped structure) with (I) rigid wall boundary conditions and (II) pressure-release boundaries. The rigid-wall (pressure-release) acoustic enclosure problem is a Neumann (Dirichlet) boundary condition problem. In both cases, eigenfunctions and eigenmodes are calculated and the shape dependence of the eigenvalue for the ground state is examined. PMID- 14758999 TI - Asymptotic solutions of the inhomogeneous Burgers equation. AB - The method of the active second harmonic suppression in resonators is investigated in this paper both analytically and numerically. The resonator is driven by a piston which vibrates with two frequencies. The first one agrees with an eigenfrequency and the second one is equal to the two times higher eigenfrequency. The phase shift of the second piston motion is 180 deg. It is known that for this case it is possible to describe generation of the higher harmonics by means of the inhomogeneous Burgers equation. This model equation was solved for stationary state analytically by a number of authors but only for ideal fluids. Unlike their solutions, new asymptotic solutions are presented here which take into account dissipative effects. The asymptotic solutions are compared with numerical ones. For study of generation higher harmonics the solutions are developed in a spectral form. PMID- 14759000 TI - Steady state unfocused circular aperture beam patterns in nonattenuating and attenuating fluids. AB - Single integral approximate formulas have been derived for the axial and lateral pressure magnitudes in the beam pattern of steady state unfocused circular flat piston sources radiating into nonattenuating and attenuating fluids. The nonattenuating formulas are shown to be highly accurate at shallow beam depths if a normalized form of the beam pattern is utilized. The axial depth of the beginning of the nonattenuated beam pattern far field is found to be at 6.41Y0. It is demonstrated that the nonattenuated lateral beam profile is represented at this and deeper depths by a Jinc function directivity term. Values of alpha and z are found that permit the attenuated axial pressure to be represented by a plane wave multiplicative attenuation factor. This knowledge should aid in the experimental design of high accuracy attenuation measurements. The shifts in depth of the principal axial pressure maxima and minima due to fluid attenuation are derived. Single integral approximate equations for the attenuated full beam pattern pressure are presented using complex Bessel functions. PMID- 14759001 TI - Spontaneous pattern formation in an acoustical resonator . AB - A dynamical system of equations describing parametric sound generation (PSG) in a dispersive large aspect ratio resonator is derived. The model generalizes previously proposed descriptions of PSG by including diffraction effects and is analogous to the model used in theoretical studies of optical parametric oscillation. A linear stability analysis of the solution below the threshold of subharmonic generation reveals the existence of a pattern forming instability, which is confirmed by numerical integration. The conditions of emergence of periodic patterns in transverse space are discussed in the acoustical context. PMID- 14759002 TI - Spherical wave propagation through inhomogeneous, anisotropic turbulence: log amplitude and phase correlations. AB - Inhomogeneity and anisotropy are intrinsic characteristics of daytime and nighttime turbulence in the atmospheric boundary layer. In the present paper, line-of-sight sound propagation through inhomogeneous, anisotropic turbulence with temperature and velocity fluctuations is considered. Starting from a parabolic equation and using the Markov approximation, formulas are derived for the correlation functions and variances of log-amplitude and phase fluctuations of a spherical sound wave. These statistical moments of a sound field are important for many practical applications in atmospheric acoustics. The derived formulas for the correlation functions and variances generalize those already known in the literature for two limiting cases: (a) homogeneous, isotropic turbulence, and (b) inhomogeneous, anisotropic turbulence with temperature fluctuations only. Furthermore, the formulas differ from those for the case of plane wave propagation. Using the derived formulas and Mann's spectral tensor of velocity fluctuations for shear-driven turbulence, the correlation functions and variances of log-amplitude and phase fluctuations are studied numerically. The results obtained clearly show that turbulence inhomogeneity and anisotropy significantly affect sound propagation in the atmosphere. PMID- 14759003 TI - The sonar equation and the definitions of propagation loss. AB - A rigorous application of the traditional definition of sonar equation terms leads to the appearance of an unexpected factor, not routinely included, equal to the ratio of the characteristic impedance at the receiver to that at the source. An omission of this factor can lead to non-negligible errors for realistic conditions. It is further argued that a gradual change in the de facto definition of propagation loss occurred between 1965 and 1980. Two alternatives to the traditional sonar equation are suggested, each using one of the two propagation loss definitions and both eliminating the unwanted impedance ratio. PMID- 14759004 TI - Tabu for matched-field source localization and geoacoustic inversion. AB - Tabu is a global optimization technique that has been very successful in operations research. In this paper, a Tabu-based method is developed for source localization and geoacoustic inversion with underwater sound data; the method relies on memory to guide the multiparameter search. Tabu is evaluated through a comparison to simulating annealing. Both methods are tested by inverting synthetic data for various numbers of unknown parameters. Tabu is found to be superior to the simulated annealing variant implemented here in terms both of accuracy and efficiency. Inversion results from the SWellEX-96 data set are also presented. PMID- 14759005 TI - Propagation of ultrasonic guided waves in lap-shear adhesive joints: case of incident a0 Lamb wave. AB - This paper deals with the propagation of ultrasonic guided waves in adhesively bonded lap-shear joints. The topic is relevant to bond inspection by ultrasonic testing. Specifically, the propagation of the lowest-order, antisymmetric a0 mode through the joint is examined. An important aspect is the mode conversion at the boundaries between the single-plate adherents and the multilayer overlap. The a0 strength of transmission is studied for three different bond states in aluminum joints, namely a fully cured adhesive bond, a poorly cured adhesive bond, and a slip bond. Theoretical predictions indicate that the dispersive behavior of the guided waves in the multilayer overlap is highly dependent on bond state. Experimental tests are conducted in lap-shear joints by a hybrid, broadband laser/air-coupled ultrasonic setup in a through-transmission configuration. The Gabor wavelet transform is employed to extract energy transmission coefficients in the 100 kHz 1.4 MHz range for the three different bond states examined. The cross-sectional mode shapes of the guided waves are shown to have a substantial role in the energy transfer through the joint. PMID- 14759006 TI - Ultrasonic interferometry for the measurement of shear velocity and attenuation in viscoelastic solids. AB - A method for the measurement of the shear properties of solid viscoelastic materials is presented. The viscoelastic material is cut into a cylindrical sample which is clamped between two rods. The transmission and reflection coefficient spectra of the fundamental torsional mode through the sample are measured by means of two pairs of piezoelectric transducers placed at the free ends of the rod-sample-rod system. Such spectra exhibit maxima and minima which occur approximately at the resonance frequencies of the free viscoelastic cylinder. Therefore, the shear velocity can be obtained by measuring the frequency interval between two consecutive maxima or minima. The shear attenuation is derived by best fitting the analytical expression of the reflection and transmission coefficients to the experimental spectra. The test is very quick to set up as the sample is simply clamped between the two rods. PMID- 14759007 TI - Local surface skimming longitudinal wave velocity and residual stress mapping. AB - Local variation in surface skimming longitudinal wave (SSLW) velocity has been measured using a scanning acoustic microscope. A very narrow width electrical impulse has been used to excite the transducer of the acoustic lens. This permits the separation of the SSLW signal from the direct reflected signal in the time domain. A simple method of measuring the time delay between the directly reflected signal and the SSLW signal at two defocuses has been utilized for the local measurement of SSLW velocity. The variation in the SSLW velocity measured over an area of the sample is scaled and presented as an image. The method has been implemented to image the variation of the SSLW velocity around a crack tip in a sample of Ti-6Al-4V. Since the SSLW velocity is known to change linearly with the stress, the SSLW velocity image is considered as a representation of the image of stress around the crack tip. Local stress variation in the same region of the crack tip is directly measured using x-ray diffraction. The SSLW velocity image is compared with the x-ray diffraction stress image. The contrast in the two images, spatial resolution, and the penetration depth into the sample of acoustic waves and x rays are discussed. PMID- 14759008 TI - Interaction of a scanning laser-generated ultrasonic line source with a surface breaking flaw. AB - The scanning laser source (SLS) technique has been proposed recently as an effective way to investigate small surface-breaking cracks. By monitoring the amplitude and frequency changes of the ultrasound generated as the SLS scans over a defect, the SLS technique has provided enhanced signal-to-noise performance compared to the traditional pitch-catch or pulse-echo ultrasonic methods. In previous work, either a point source or a short line source was used for generation of ultrasound. The resulting Rayleigh wave was typically bipolar in nature. In this paper, a scanning laser line source (SLLS) technique using a true thermoelastic line source (which leads to generation of monopolar surface waves) is demonstrated experimentally and through numerical simulation. Experiments are performed using a line-focused Nd:YAG laser and interferometric detection. For the numerical simulation, a hybrid model combining a mass-spring lattice method (MSLM) and a finite difference method (FDM) is used. As the SLLS is scanned over a surface-breaking flaw, it is shown both experimentally and numerically that the monopolar Rayleigh wave becomes bipolar, dramatically indicating the presence of the flaw. PMID- 14759009 TI - Optimization of the performance of the sandwich piezoelectric ultrasonic transducer. AB - The resonance and antiresonance frequency, the effective electromechanical coupling coefficient, and the mechanical quality factor of a sandwich piezoelectric ultrasonic transducer are studied and optimized. The effect of the thickness of thick piezoelectric element electrodes on the transducer performance is analyzed. The effect of the length and position of the piezoelectric elements in the transducer is also studied. It is shown that, although using thick electrodes is beneficial for releasing heat produced by the piezoelectric elements, the effective electromechanical coupling coefficient and the mechanical quality factor are reduced. The length and the position of the piezoelectric elements affect the performances of the transducer. Increasing the length of the piezoelectric elements decreases the mechanical quality factor, but the effective electromechanical coupling coefficient increases. When the length reaches a certain value, the effective electromechanical coupling coefficient reaches a maximum value. When the piezoelectric elements are located at the geometrical center or the displacement node, the effective electromechanical coupling coefficient and the mechanical quality factor are maximized. PMID- 14759010 TI - Nonperturbing measurements of spatially distributed underwater acoustic fields using a scanning laser Doppler vibrometer. AB - Localized changes in the density of water induced by the presence of an acoustic field cause perturbations in the localized refractive index. This relationship has given rise to a number of nonperturbing optical metrology techniques for recording measurement parameters from underwater acoustic fields. A method that has been recently developed involves the use of a Laser Doppler Vibrometer (LDV) targeted at a fixed, nonvibrating, plate through an underwater acoustic field. Measurements of the rate of change of optical pathlength along a line section enable the identification of the temporal and frequency characteristics of the acoustic wave front. This approach has been extended through the use of a scanning LDV, which facilitates the measurement of a range of spatially distributed parameters. A mathematical model is presented that relates the distribution of pressure amplitude and phase in a planar wave front with the rate of change of optical pathlength measured by the LDV along a specifically orientated laser line section. Measurements of a 1 MHz acoustic tone burst generated by a focused transducer are described and the results presented. Graphical depictions of the acoustic power and phase distribution recorded by the LDV are shown, together with images representing time history during the acoustic wave propagation. PMID- 14759011 TI - Surface fields on nonseparable geometries. AB - The field on the surface of a structure having nonseparable geometry may be expanded in a set of eigenfunctions suitable for that particular geometry. The eigenfunctions are computed numerically using the boundary elements method to compute a matrix that relates the field on the surface of the structure to its normal derivative at vanishing frequency and diagonalizing the matrix. The field on the surface of a finite cylinder with hemisphical end-caps is expanded in such a set of eigenfunctions for both a simple cylinder and a cylinder containing frames. Results for the framed cylinder is compared to Fourier decomposition of the field on the cylindrical portion of the structure. PMID- 14759012 TI - Detection and localization of inclusions in plates using inversion of point actuated surface displacements. AB - A numerical simulation is carried out demonstrating the use of plate surface vibration measurements for detecting and locating inclusions within the structure. A finite element code is used to calculate normal surface displacement for both steel and mortar plates subjected to a monochromatic point force. The data is generated for the homogeneous plate and the identical plate within which exists a small rectangular inclusion. It is observed that when the elastic modulus of the inclusion is orders of magnitude lower than the base material, resonances of the inclusion produce large local displacements that are readily observed in the raw displacement data. For more modest moduli differences, there are no such directly observable effects. In this case, three inverse algorithms are used to process the displacement data. The first two are local inversion techniques that each yield a spatial map of the elastic modulus normalized by density. These algorithms successfully detect and localize the inclusion based on its modulus difference from that of the base plate. The third technique uses a form of the inhomogeneous equation of motion to obtain the induced force distribution connected with the inclusion. The spatial mapping of this force also successfully detects and localizes the inclusion. PMID- 14759013 TI - Hybrid near-field acoustic holography. AB - Hybrid near-field acoustical holography (NAH) is developed for reconstructing acoustic radiation from an arbitrary object in a cost-effective manner. This hybrid NAH is derived from a modified Helmholtz equation least squares (HELS) formula that expands the acoustic pressure in terms of outgoing and incoming waves. The expansion coefficients are determined by solving an overdetermined linear system of equations obtained by matching the assumed-form solution to measured acoustic pressures through the least squares. Measurements are taken over a conformal surface around a source at close range so that the evanescent waves can be captured. Next, the modified HELS is utilized to regenerate as much acoustic pressures on the conformal surface as necessary and take them as input to the Helmholtz integral formulation implemented numerically by boundary element method (BEM). The acoustic pressures and normal velocities on the source surface are reconstructed by using a modified Tikhnov regularization (TR) with its regularization parameter determined by generalized cross validation (GCV) method. Results demonstrate that this hybrid NAH combines the advantages of HELS and inverse BEM. This is because a majority of the input data are regenerated but not measured, thus the efficiency of reconstruction is greatly enhanced. Meanwhile, the accuracy of reconstruction is ensured by the Helmholtz integral theory and modified TR together with GCV method, provided that HELS converges fast enough on the measurement surface. Numerical examples of reconstructing acoustic quantities on the surface of a simplified engine block are demonstrated. [Work supported by NSF.] PMID- 14759014 TI - Sound transmission characteristics of Tee-junctions and the associated length corrections. AB - The sound transmission characteristics of a Tee-junction formed by a sidebranch and an infinitely long duct are investigated numerically using the finite element method. The associated corrections of the branch length and the upstream duct length are also discussed in detail. The types of branch resonance that result in strong or weak sound transmission across the junction are determined and their effects on the length corrections examined. Results suggest that the type of sidebranch, the branch width, the branch length, and the order and the form of the resonance affect more significantly the length corrections of the duct section. The excitation of nonplanar higher branch modes gives rise to rapid increase in the duct length corrections and also results in lower sound transmission. PMID- 14759015 TI - Application of theoretical modeling to multichannel active control of cooling fan noise. AB - Multichannel active control has been applied to the global reduction of tonal noise from a cooling fan. In order to achieve consistent far-field attenuation of multiple harmonics of the blade passage frequency (BPF) of the fan, an analytical model has been applied to the control system in order to determine appropriate transducer configurations. The results of the modeling show that the additional global reduction possible by locating acoustically compact secondary sources coplanar with a compact primary source rapidly lessens as the number of symmetrically placed sources is increased beyond three. Furthermore, the model suggests that there are locations in the extreme near field of the sources that can be considered ideal for the minimization of far-field radiated power. Experiments carried out show that a four-channel control system is more effective than a two-channel system at achieving far-field attenuations, especially at the higher harmonics of the BPF for the fan tested. In addition, greater far-field mean-square pressure attenuations are achieved with the error microphones located along the calculated ideal regions than for nonideal placement. PMID- 14759016 TI - Nonoccupational noise: exposures associated with routine activities. AB - Efforts to characterize nonoccupational noise exposures have focused primarily on infrequent, episodic events. Few studies have assessed noise levels resulting from routine daily activities. In the current study, 112 construction workers wore datalogging noise dosimeters and simultaneously completed activity logs during two phases of data collection. The 81 subjects monitored in phase 1 received logs listing numerous preselected occupational and nonoccupational activities, while the 31 subjects monitored in phase 2 used free-field logs and reported nonoccupational activities in greater detail. Nearly all of the 221,439 1-min intervals of nonoccupational L(eq) level and activity reporting were below 70 dBA; only a small percentage exceeded 80 dBA. The primary contributor to nonoccupational noise exposure was traveling in a car or bus, while time at home contributed the least. One hundred seventy 24-h L(eq) levels were computed from the 1-min noise level data. The percentage of phase 2 workday L(eq(24)) levels which exceeded 80 dBA was higher than that of the nonworkday levels. The mean L(eq(24)) level of phase 2 workdays was higher than that of nonworkdays, and the difference was statistically significant. Routine nonoccupational noise exposures contributed much less to total noise dose than occupational exposures in the subjects evaluated. PMID- 14759017 TI - Nonlinear synthesis of input signals in ultrasonic experimental setups. AB - A method is proposed to find the digital input signals to enter in nonlinear acoustical systems (power amplifiers, transducers, etc.) in order to obtain desired arbitrary response signals. The searched input signals are found by performing a Monte Carlo search guided by a simulated annealing process applied to a hidden model with a small number of parameters. The physical system is actually used in the optimization procedure, in a real-time manner, so that no theoretical model of the system response is required. The main aspects of the algorithm are described and illustrated with several examples. PMID- 14759018 TI - Uncertainties associated with parameter estimation in atmospheric infrasound arrays. AB - This study describes a method for determining the statistical confidence in estimates of direction-of-arrival and trace velocity stemming from signals present in atmospheric infrasound data. It is assumed that the signal source is far enough removed from the infrasound sensor array that a plane-wave approximation holds, and that multipath and multiple source effects are not present. Propagation path and medium inhomogeneities are assumed not to be known at the time of signal detection, but the ensemble of time delays of signal arrivals between array sensor pairs is estimable and corrupted by uncorrelated Gaussian noise. The method results in a set of practical uncertainties that lend themselves to a geometric interpretation. Although quite general, this method is intended for use by analysts interpreting data from atmospheric acoustic arrays, or those interested in designing and deploying them. The method is applied to infrasound arrays typical of those deployed as a part of the International Monitoring System of the Comprehensive Nuclear-Test-Ban Treaty Organization. PMID- 14759019 TI - Wideband time-reversal imaging of an elastic target in an acoustic waveguide. AB - Time-reversal is addressed for imaging elastic targets situated in an acoustic waveguide. It is assumed that the target-sensor range is large relative to the channel depth. We investigate the theory of wideband time-reversal imaging of an extended elastic target, for which the target dimensions are large relative to the principal wavelengths. When performing time-reversal imaging one requires a forward model for propagation through the channel, and the quality of the resulting image may be used as a measure of the match between the modeled and actual (measured) channel parameters. It is demonstrated that the channel parameters associated with a given measurement may be determined via a genetic algorithm (GA) search in parameter space, employing a cost function based on the time-reversal image quality. Example GA channel-parameter-inversion results and imagery are presented for measured at-sea data. PMID- 14759020 TI - Inversion of guided-wave dispersion data with application to borehole acoustics. AB - The problem of inferring unknown geometry and material parameters of a waveguide model from noisy samples of the associated modal dispersion curves is considered. In a significant reduction of the complexity of a common inversion methodology, the inner of two nested iterations is eliminated: The approach described does not employ explicit fitting of the data to computed dispersion curves. Instead, the unknown parameters are adjusted to minimize a cost function derived directly from the determinant of the boundary condition system matrix. This results in an efficient inversion scheme that, in the case of noise-free data, yields exact results. Multimode data can be simultaneously processed without extra complications. Furthermore, the inversion scheme can accommodate an arbitrary number of unknown parameters, provided that the data have sufficient sensitivity to these parameters. As an important application, we consider the sonic guidance condition for a fluid-filled borehole in an elastic, homogeneous, and isotropic rock formation for numerical forward and inverse dispersion analysis. We investigate numerically the parametric inversion with errors in the model parameters and the influence of bandwidth and noise, and examine the cases of multifrequency and multimode data, using simulated flexural and Stoneley dispersion data. PMID- 14759021 TI - Representation of concurrent acoustic objects in primary auditory cortex. AB - Auditory scene analysis involves the simultaneous grouping and parsing of acoustic data into separate mental representations (i.e., objects). Over two experiments, we examined the sequence of neural processes underlying concurrent sound segregation by means of recording of human middle latency auditory evoked responses. Participants were presented with complex sounds comprising several harmonics, one of which could be mistuned such that it was not an integer multiple of the fundamental frequency. In both experiments, Na (approximately 22 ms) and Pa (approximately 32 ms) waves were reliably generated for all classes of stimuli. For stimuli with a fundamental frequency of 200 Hz, the mean Pa amplitude was significantly larger when the third harmonic was mistuned by 16% of its original value, relative to when it was tuned. The enhanced Pa amplitude was related to an increased likelihood in reporting the presence of concurrent auditory objects. Our results are consistent with a low-level stage of auditory scene analysis in which acoustic properties such as mistuning act as preattentive segregation cues that can subsequently lead to the perception of multiple auditory objects. PMID- 14759022 TI - Cuing effects for informational masking. AB - The detection of a tone added to a random-frequency, multitone masker can be very poor even when the maskers have little energy in the frequency region of the signal. This paper examines the effects of adding a pretrial cue to reduce uncertainty for the masker or the signal. The first two experiments examined the effect of cuing a fixed-frequency signal as the number of masker components and presentation methods were manipulated. Cue effectiveness varied across observers, but could reduce thresholds by as much as 20 dB. Procedural comparisons indicated observers benefited more from having two masker samples to compare, with or without a signal cue, than having a single interval with one masker sample and a signal cue. The third experiment used random-frequency signals and compared no cue, signal-cue, and masker-cue conditions, and also systematically varied the time interval between cue offset and trial onset. Thresholds with a cued random frequency signal remained higher than for a cued fixed-frequency signal. For time intervals between the cue and trial of 50 ms or longer, thresholds were approximately the same with a signal or a masker cue and lower than when there was no cue. Without a cue or with a masker cue, analyses of possible decision strategies suggested observers attended to the potential signal frequencies, particularly the highest signal frequency. With a signal cue, observers appeared to attend to the frequency of the subsequent signal. PMID- 14759023 TI - Within-ear and across-ear interference in a dichotic cocktail party listening task: effects of masker uncertainty. AB - Increases in masker variability have been shown to increase the effects of informational masking in non-speech listening tasks, but relatively little is known about the influence that masker uncertainty has on the informational components of speech-on-speech masking. In this experiment, listeners were asked to extract information from a target phrase that was presented in their right ear while ignoring masking phrases that were presented in the same ear as the target phrase and in the ear opposite the target phrase. The level of masker uncertainty was varied by holding constant or "freezing" the talkers speaking the masking phrases, the semantic content used in the masking phrases, or both the talkers and the semantic content in the masking phrases within each block of 120 trials. The results showed that freezing the semantic content of the masking phrase in the target ear was the only reduction in masker uncertainty that ever resulted in a significant improvement in performance. Providing feedback after each trial improved performance overall, but did not prevent the listeners from making incorrect responses that matched the content of the frozen target-ear masking phrase. However, removing the target-ear contents corresponding to the masking phrase from the response set resulted in a dramatic improvement in performance. This suggests that the listeners were generally able to understand both of the phrases presented to the target ear, and that their incorrect responses in the task were almost entirely a result of their inability to determine which words were spoken by the target talker. PMID- 14759024 TI - Development of a new standard laboratory protocol for estimation of the field attenuation of hearing protection devices: sample size necessary to provide acceptable reproducibility. AB - The mandate of ASA Working Group S12/WG11 has been to develop "laboratory and/or field procedure(s) that yield useful estimates of field performance" of hearing protection devices (HPDs). A real-ear attenuation at threshold procedure was selected, devised, tested for one earmuff and three earplugs via an interlaboratory study involving five laboratories and 147 subjects, and incorporated into a new standard that was approved in 1997 [Royster et al., "Development of a new standard laboratory protocol for estimating the field attenuation of hearing protection devices. Part I. Research of Working Group 11, Accredited Standards Committee S 12, Noise," J. Acoust. Soc. Am. 99, 1506-1526; ANSI, S12.6-1997, "American National Standard method for measuring real-ear attenuation of hearing protectors" (American National Standards Institute, New York, 1997)]. The subject-fit methodology of ANSI S12.6-1997 relies upon listeners who are audiometrically proficient, but inexperienced in the use of HPDs. Whenever a new method is adopted, it is important to know the effects of variability on the power of the measurements. In evaluation of protector noise reduction determined by experimenter-fit, informed-user-fit, and subject-fit methods, interlaboratory reproducibility was found to be best for the subject-fit method. Formulas were derived for determining the minimum detectable difference between attenuation measurements and for determining the number of subjects necessary to achieve a selected level of precision. For a precision of 6 dB, the study found that the minimum number of subjects was 4 for the Bilsom UF-1 earmuff, 10 for the E.A.R Classic earplug, 31 for the Willson EP100 earplug, and 22 for the PlasMed V-51R earplug. PMID- 14759025 TI - Separation of concurrent broadband sound sources by human listeners. AB - The effect of spatial separation on the ability of human listeners to resolve a pair of concurrent broadband sounds was examined. Stimuli were presented in a virtual auditory environment using individualized outer ear filter functions. Subjects were presented with two simultaneous noise bursts that were either spatially coincident or separated (horizontally or vertically), and responded as to whether they perceived one or two source locations. Testing was carried out at five reference locations on the audiovisual horizon (0 degrees, 22.5 degrees, 45 degrees, 67.5 degrees, and 90 degrees azimuth). Results from experiment 1 showed that at more lateral locations, a larger horizontal separation was required for the perception of two sounds. The reverse was true for vertical separation. Furthermore, it was observed that subjects were unable to separate stimulus pairs if they delivered the same interaural differences in time (ITD) and level (ILD). These findings suggested that the auditory system exploited differences in one or both of the binaural cues to resolve the sources, and could not use monaural spectral cues effectively for the task. In experiments 2 and 3, separation of concurrent noise sources was examined upon removal of low-frequency content (and ITDs), onset/offset ITDs, both of these in conjunction, and all ITD information. While onset and offset ITDs did not appear to play a major role, differences in ongoing ITDs were robust cues for separation under these conditions, including those in the envelopes of high-frequency channels. PMID- 14759026 TI - A model of acoustic interspeaker variability based on the concept of formant cavity affiliation. AB - A method is proposed to model the interspeaker variability of formant patterns for oral vowels. It is assumed that this variability originates in the differences existing among speakers in the respective lengths of their front and back vocal-tract cavities. In order to characterize, from the spectral description of the acoustic speech signal, these vocal-tract differences between speakers, each formant is interpreted, according to the concept of formant-cavity affiliation, as a resonance of a specific vocal-tract cavity. Its frequency can thus be directly related to the corresponding cavity length, and a transformation model can be proposed from a speaker A to a speaker B on the basis of the frequency ratios of the formants corresponding to the same resonances. In order to minimize the number of sounds to be recorded for each speaker in order to carry out this speaker transformation, the frequency ratios are exactly computed only for the three extreme cardinal vowels [i, a, u] and they are approximated for the remaining vowels through an interpolation function. The method is evaluated through its capacity to transform the (F1,F2) formant patterns of eight oral vowels pronounced by five male speakers into the (F1,F2) patterns of the corresponding vowels generated by an articulatory model of the vocal tract. The resulting formant patterns are compared to those provided by normalization techniques published in the literature. The proposed method is found to be efficient, but a number of limitations are also observed and discussed. These limitations can be associated with the formant-cavity affiliation model itself or with a possible influence of speaker-specific vocal-tract geometry in the cross sectional direction, which the model might not have taken into account. PMID- 14759027 TI - Vowel normalization for accent: an investigation of best exemplar locations in northern and southern British English sentences. AB - Two experiments investigated whether listeners change their vowel categorization decisions to adjust to different accents of British English. Listeners from different regions of England gave goodness ratings on synthesized vowels embedded in natural carrier sentences that were spoken with either a northern or southern English accent. A computer minimization algorithm adjusted F1, F2, F3, and duration on successive trials according to listeners' goodness ratings, until the best exemplar of each vowel was found. The results demonstrated that most listeners adjusted their vowel categorization decisions based on the accent of the carrier sentence. The patterns of perceptual normalization were affected by individual differences in language background (e.g., whether the individuals grew up in the north or south of England), and were linked to the changes in production that speakers typically make due to sociolinguistic factors when living in multidialectal environments. PMID- 14759028 TI - Acoustic properties of naturally produced clear speech at normal speaking rates. AB - Sentences spoken "clearly" are significantly more intelligible than those spoken "conversationally" for hearing-impaired listeners in a variety of backgrounds [Picheny et al., J. Speech Hear. Res. 28, 96-103 (1985); Uchanski et al., ibid. 39, 494-509 (1996); Payton et al., J. Acoust. Soc. Am. 95, 1581-1592 (1994)]. While producing clear speech, however, talkers often reduce their speaking rate significantly [Picheny et al., J. Speech Hear. Res. 29, 434-446 (1986); Uchanski et al., ibid. 39, 494-509 (1996)]. Yet speaking slowly is not solely responsible for the intelligibility benefit of clear speech (over conversational speech), since a recent study [Krause and Braida, J. Acoust. Soc. Am. 112, 2165-2172 (2002)] showed that talkers can produce clear speech at normal rates with training. This finding suggests that clear speech has inherent acoustic properties, independent of rate, that contribute to improved intelligibility. Identifying these acoustic properties could lead to improved signal processing schemes for hearing aids. To gain insight into these acoustical properties, conversational and clear speech produced at normal speaking rates were analyzed at three levels of detail (global, phonological, and phonetic). Although results suggest that talkers may have employed different strategies to achieve clear speech at normal rates, two global-level properties were identified that appear likely to be linked to the improvements in intelligibility provided by clear/normal speech: increased energy in the 1000-3000-Hz range of long-term spectra and increased modulation depth of low frequency modulations of the intensity envelope. Other phonological and phonetic differences associated with clear/normal speech include changes in (1) frequency of stop burst releases, (2) VOT of word-initial voiceless stop consonants, and (3) short-term vowel spectra. PMID- 14759029 TI - Performance of time- and frequency-domain binaural beamformers based on recorded signals from real rooms. AB - Extraction of a target sound source amidst multiple interfering sound sources is difficult when there are fewer sensors than sources, as is the case for human listeners in the classic cocktail-party situation. This study compares the signal extraction performance of five algorithms using recordings of speech sources made with three different two-microphone arrays in three rooms of varying reverberation time. Test signals, consisting of two to five speech sources, were constructed for each room and array. The signals were processed with each algorithm, and the signal extraction performance was quantified by calculating the signal-to-noise ratio of the output. A frequency-domain minimum-variance distortionless-response beamformer outperformed the time-domain based Frost beamformer and generalized sidelobe canceler for all tests with two or more interfering sound sources, and performed comparably or better than the time domain algorithms for tests with one interfering sound source. The frequency domain minimum-variance algorithm offered performance comparable to that of the Peissig-Kollmeier binaural frequency-domain algorithm, but with much less distortion of the target signal. Comparisons were also made to a simple beamformer. In addition, computer simulations illustrate that, when processing speech signals, the chosen implementation of the frequency-domain minimum variance technique adapts more quickly and accurately than time-domain techniques. PMID- 14759030 TI - Methodology for rheological testing of engineered biomaterials at low audio frequencies. AB - A commercial rheometer (Bohlin CVO120) was used to mechanically test materials that approximate vocal-fold tissues. Application is to frequencies in the low audio range (20-150 Hz). Because commercial rheometers are not specifically designed for this frequency range, a primary problem is maintaining accuracy up to (and beyond) the mechanical resonance frequency of the rotating shaft assembly. A standard viscoelastic material (NIST SRM 2490) has been used to calibrate the rheometric system for an expanded frequency range. Mathematically predicted response curves are compared to measured response curves, and an error analysis is conducted to determine the accuracy to which the elastic modulus and the shear modulus can be determined in the 20-150-Hz region. Results indicate that the inertia of the rotating assembly and the gap between the plates need to be known (or determined empirically) to a high precision when the measurement frequency exceeds the resonant frequency. In addition, a phase correction is needed to account for the magnetic inertia (inductance) of the drag cup motor. Uncorrected, the measured phase can go below the theoretical limit of -pi. This can produce large errors in the viscous modulus near and above the resonance frequency. With appropriate inertia and phase corrections, +/- 10% accuracy can be obtained up to twice the resonance frequency. PMID- 14759031 TI - Reverberation and frequency attenuation in forests--implications for acoustic communication in animals. AB - Rates of reverberative decay and frequency attenuation are measured within two Australian forests. In particular, their dependence on the distance between a source and receiver, and the relative heights of both, is examined. Distance is always the most influential of these factors. The structurally denser of the forests exhibits much slower reverberative decay, although the frequency dependence of reverberation is qualitatively similar in the two forests. There exists a central range of frequencies between 1 and 3 kHz within which reverberation varies relatively little with distance. Attenuation is much greater within the structurally denser forest, and in both forests it generally increases with increasing frequency and distance, although patterns of variation differ between the two forests. Increasing the source height generally reduces reverberation, while increasing the receiver height generally reduces attenuation. These findings have considerable implications for acoustic communication between inhabitants of these forests, particularly for the perching behaviors of birds. Furthermore, this work indicates the ease with which the general acoustic properties of forests can be measured and compared. PMID- 14759032 TI - Sex differences in the acoustic structure of vowel-like grunt vocalizations in baboons and their perceptual discrimination by baboon listeners. AB - This study quantifies sex differences in the acoustic structure of vowel-like grunt vocalizations in baboons (Papio spp.) and tests the basic perceptual discriminability of these differences to baboon listeners. Acoustic analyses were performed on 1028 grunts recorded from 27 adult baboons (11 males and 16 females) in southern Africa, focusing specifically on the fundamental frequency (F0) and formant frequencies. The mean F0 and the mean frequencies of the first three formants were all significantly lower in males than they were in females, more dramatically so for F0. Experiments using standard psychophysical procedures subsequently tested the discriminability of adult male and adult female grunts. After learning to discriminate the grunt of one male from that of one female, five baboon subjects subsequently generalized this discrimination both to new call tokens from the same individuals and to grunts from novel males and females. These results are discussed in the context of both the possible vocal anatomical basis for sex differences in call structure and the potential perceptual mechanisms involved in their processing by listeners, particularly as these relate to analogous issues in human speech production and perception. PMID- 14759034 TI - Mode properties of an external-cavity laser with Gaussian gain. AB - We analyze the mode properties of a laser with a Gaussian gain profile by using the beam propagation method. The resonance properties of the Petermann K factor and the M2 beam quality are shown to be related in the vicinity of degenerate cavity geometries. K is unity for a confocal cavity, even under conditions with strong gain guiding, while M2 is a maximum. PMID- 14759033 TI - Determination of West Indian manatee vocalization levels and rate. AB - The West Indian manatee (Trichechus manatus latirostris) has become endangered partly because of a growing number of collisions with boats. A system to warn boaters of the presence of manatees, based upon the vocalizations of manatees, could potentially reduce these boat collisions. The feasibility of this warning system would depend mainly upon two factors: the rate at which manatees vocalize and the distance in which the manatees can be detected. The research presented in this paper verifies that the average vocalization rate of the West Indian manatee is approximately one to two times per 5-min period. Several different manatee vocalization recordings were broadcast to the manatees and their response was observed. It was found that during the broadcast periods, the vocalization rates for the manatees increased substantially when compared with the average vocalization rates during nonbroadcast periods. An array of four hydrophones was used while recording the manatees. This allowed for position estimation techniques to be used to determine the location of the vocalizing manatee. Knowing the position of the manatee, the source level was determined and it was found that the mean source level of the manatee vocalizations is approximately 112 dB (re 1 microPa) @ 1 m. PMID- 14759035 TI - Off-resonant defocus-contrast imaging of cold atoms. AB - We demonstrate the retrieval of column-density images of cold atoms, using a noninterferometric phase-recovery technique based on a single off-resonant and defocused intensity image. The quantitative column density is retrieved via Fourier inversion and remains robust with respect to detuning and defocus. The technique offers excellent prospects for simple, nondestructive imaging of atoms in magnetic and optical traps and condensates. PMID- 14759036 TI - First- and second-order statistics of optical near fields. AB - The statistical properties of the intensity in close proximity to highly scattering, randomly inhomogeneous media are investigated. Whereas the intensity probability density function obeys the same law irrespective of the distance z from the interface, the second-order intensity correlation length changes for distances smaller than the wavelength. Contrary to predictions of the conventional coherence theory, the corresponding field correlation length can be smaller than the wavelength of light. PMID- 14759037 TI - Propagation-invariant vectorial Bessel beams obtained by use of quantized Pancharatnam-Berry phase optical elements. AB - Propagation-invariant vectorial Bessel beams with linearly polarized axial symmetry based on quantized Pancharatnam-Berry phase optical elements are described. The geometric phase is formed through the use of discrete computer generated space-variant subwavelength dielectric gratings. We have verified the polarization properties of our elements for laser radiation at 10.6-microm wavelength and also demonstrated propagation-invariant, controlled rotation of a propeller-shaped intensity pattern through the simple rotation of a polarizer. PMID- 14759038 TI - Generation of 40-GHz control signals from flag pulses for switching all-optical gates for use with optical packets. AB - We demonstrate an all-optical circuit capable of generating 40-GHz control signals from flag pulses that can be used to define the switching state of all optical gates for use with optical packets. The circuit comprises a Fabry-Perot filter and a semiconductor optical amplifier, and with a single pulse it can generate 12 control pulses with 0.64-dB amplitude modulation. With two and three flag pulses the number of control pulses becomes 36 and 54, respectively. PMID- 14759039 TI - Fabrication of a tilted fiber Bragg grating with a designed reflection spectrum profile. AB - A method of fabricating a tilted fiber Bragg grating (TFBG) with a designed reflection spectrum profile is proposed and demonstrated experimentally. In a reflective TFBG the central wavelength and the reflectivity are dependent on the tilt angles. Controlling the rate at which the tilt angle is continuously varied and the exposure dose in the UV-light writing process permits accurate control of the bandwidth and reflectivity of the grating. The experimental result agrees well with TFBG theory and calculation. PMID- 14759040 TI - Gas-phase temperature measurement in the vaporizing spray of a gasoline direct injection injector by use of pure rotational coherent anti-Stokes Raman scattering. AB - Pure rotational coherent anti-Stokes Raman spectroscopy is applied for quantitative gas-phase temperature measurements in the vaporizing spray of an automotive fuel injector. Interferences from elastically scattered stray light are greatly reduced by use of a polarization technique and spectral filtering in a double monochromator. The applicability of this technique to probing low temperature sprays is successfully demonstrated. PMID- 14759041 TI - Phase-locked, erbium-fiber-laser-based frequency comb in the near infrared. AB - A phase-locked frequency comb in the near infrared is demonstrated with a mode locked, erbium-doped, fiber laser whose output is amplified and spectrally broadened in dispersion-flattened, highly nonlinear optical fiber to span from 1100 to >2200 nm. The supercontinuum output comprises a frequency comb with a spacing set by the laser repetition rate and an offset by the carrier-envelope offset frequency, which is detected with the standard f-to-2f heterodyne technique. The comb spacing and offset frequency are phase locked to a stable rf signal with a fiber stretcher in the laser cavity and by control of the pump laser power, respectively. This infrared comb permits frequency metrology experiments in the near infrared in a compact, fiber-laser-based system. PMID- 14759042 TI - Noninvasive optical imaging by speckle ensemble. AB - We propose a new method imaging through scattering media. An object hidden between two biological tissues (chicken breast) is reconstructed from any speckled images obtained from the output of a multichannel optical imaging system. The effect of multiple imaging is achieved with a microlens array. Each lens is the array projects a different speckled image onto a digital camera. The set of speckled images from the entire array is first shifted to a common center and then accumulated into a single average picture. PMID- 14759043 TI - Reconstructing chromosphere concentration images directly by continuous-wave diffuse optical tomography. AB - We present an algorithm to reconstruct chromosphere concentration images directly rather than following the traditional two-step process of reconstructing wavelength-dependent absorption coefficient images and then calculating chromosphere concentration images. This procedure imposes prior spectral information into the image reconstruction that results in a dramatic improvement in the image contrast-to-noise ratio of better than 100%. We demonstrate this improvement with simulations and a dynamic blood phantom experiment. PMID- 14759044 TI - Observation of transverse instabilities in optically induced lattices. AB - We study experimentally the Bloch-wave instabilities in optically induced photonic lattices. We reveal two different instability scenarios associated with either the transverse modulational instability of a single Bloch wave or the nonlinear interband coupling between different Bloch waves. We show that the transverse instability is greatly enhanced in the induced lattice in comparison with homogeneous media. PMID- 14759045 TI - Compact, synchronously diode-pumped tunable fiber Raman source of subpicosecond solitons around 1.6 microm. AB - A compact and simple all-fiber subpicosecond soliton source is demonstrated that employs synchronous Raman amplification in dispersion-shifted fiber from a gain switched 1536-nm laser diode pump. Tuning between 1620 and 1660 nm is obtained by electronic variation of the gain-switching frequency around 54.47 MHz, and solitons as short as 400 fs are achieved. This configuration could form the basis of cheap, compact, and tunable femtosecond-pulse sources. PMID- 14759046 TI - Linear and nonlinear optical properties of Ag-As-Se chalcogenide glasses for all optical switching. AB - We prepared Ag(x)(As0.4Se0.6)(100-x) chalcogenide glasses by a melt-quenching method and measured their linear and nonlinear optical properties to evaluate their potential applications to all-optical ultrafast switching devices. Their nonlinear refraction and absorption were measured by the Z-scan method at 1.05 microm. The addition of Ag to As2Se3 glass led to an increase in the nonlinear refractive index without introducing an increase in the nonlinear absorption coefficient. The glass with a Ag content of x = 20 at. % revealed high nonlinearity ranging from 2000 to 27,000 times that of fused silica, depending on the incident optical intensity. PMID- 14759047 TI - Anisotropic diffraction and elliptic discrete solitons in two-dimensional waveguide arrays. AB - We demonstrate that both the linear (diffraction) and the nonlinear dynamics of two-dimensional waveguide arrays are considerably more complex and versatile than their one-dimensional counterparts. The discrete diffraction properties of these arrays can be effectively altered, depending on the propagation Bloch k-vector within the first Brillouin zone of the lattice. In general, this diffraction behavior is anisotropic and therefore permits the existence of a new class of discrete elliptic solitons in the nonlinear regime. PMID- 14759048 TI - Adaptive control of femtosecond soliton self-frequency shift in fibers. AB - The Raman shift of a subpicosecond soliton in 100 m of fiber is controlled adaptively by pulse shaping before launching into the fiber. We use a deformable mirror-based shaper to control the spectral phase of the input pulse. PMID- 14759049 TI - Picosecond laser for performance of efficient nonlinear spectroscopy from 10 to 21 microm. AB - Laser tunability from 10 to 21 microm is obtained by use of an optical parametric oscillator based on a KTP crystal followed by a difference-frequency stage with a CdSe crystal. An all-solid-state picosecond Nd:YAG oscillator mode locked by a frequency-doubling nonlinear mirror is used for synchronous pumping. PMID- 14759050 TI - Optical spatial solitons supported by photoisomerization nonlinearity in a polymer. AB - We present a theory for a new type of optical spatial soliton that is based on the angle hole burning mechanism of photoisomerization in some polymers. We predict that the photoisomerization nonlinearity can support steady-state dark and bright spatial solitons in the polymer. We also discuss the dependence of the FWHM of the spatial soliton on wavelengths of the background beams and on the ratio of the intensity of the background beam to that of the signal beam. PMID- 14759051 TI - Soliton transverse instabilities in anisotropic nonlocal self-focusing media. AB - We study, both numerically and experimentally, the transverse modulational instability of spatial stripe solitons in anisotropic nonlocal photorefractive media. We demonstrate that the instability scenarios depend strongly on the stripe orientation, but the anisotropy-induced features are largely suppressed for spatial solitons created by self-trapping of partially incoherent light. PMID- 14759052 TI - Coherent interactions of dissipative spatial solitons. AB - We report observation of the interaction between two coherent dissipative spatial solitons in a periodically patterned semiconductor optical amplifier with power levels of tens of milliwatts. The interactions are nonlocal and phase dependent and exhibit surprising features, such as soliton birth. The experimental results are in good agreement with the numerical simulations. PMID- 14759053 TI - Attraction of nonlocal dark optical solitons. AB - We study the formation and interaction of spatial dark optical solitons in materials with a nonlocal nonlinear response. We show that unlike in local materials, where dark solitons typically repel, the nonlocal nonlinearity leads to a long-range attraction and formation of stable bound states of dark solitons. PMID- 14759054 TI - Gap soliton dynamics in a nonuniform resonant structure. AB - A model for the propagation of coherent pulses along a one-dimensional, resonantly absorbing Bragg grating that includes localized inhomogeneous population inversion at its center is presented. The long-range coupling between the optical field and resonant atoms allows for controllable trapping of a gap soliton by the local inversion, thus opening new opportunities for control of signal transmission and localization of light. PMID- 14759055 TI - Cascaded wavelength conversions based on sum-frequency generation and difference frequency generation. AB - A novel wavelength conversion scheme based on cascaded sum-frequency generation (SFG) and difference-frequency generation (DFG) is proposed and demonstrated in a MgO-doped LiNbO3 quasi-phase-matched waveguide. In this scheme, two pump wavelengths are set outside the communication band. It is shown that the same conversion efficiency can be achieved by use of two pump sources with lower output power (P1, P2) in this scheme compared with the conventional cascaded wavelength conversion technique based on second-harmonic generation and DFG with a single higher-power pump beam (P = P1 + P2) that is due to the use of a larger SFG nonlinear coefficient. The results significantly influence the selection of a suitable nonlinear interaction scheme for practical applications. PMID- 14759056 TI - Independent phase and amplitude control of a laser beam by use of a single-phase only spatial light modulator. AB - A phase-only spatial light modulator is used in conjunction with a spatial filter to provide independent control of the phase and amplitude of a laser beam. Continuous amplitude modulation of the beam is achieved with a resolution relevant to beam shaping of high-energy laser beams. Amplitude beam correction in a closed loop is demonstrated. PMID- 14759057 TI - Ultrafast holographic Stokesmeter for polarization imaging in real time. AB - We propose an ultrafast holographic Stokesmeter using a volume holographic substrate with two sets of two orthogonal gratings to identify all four Stokes parameters of the input beam. We derive the Mueller matrix of the proposed architecture and determine the constraints necessary for reconstructing the complete Stokes vector. The speed of this device is determined primarily by the channel spectral bandwidth (typically 100 GHz), corresponding to a few picoseconds. PMID- 14759058 TI - Zero-birefringence photosensitive polyimides for optical waveguides. AB - Zero-birefringence (delta n < 1 x 10(-6)) photosensitive fluorinated polyimides are synthesized for use in polymer optical waveguides. To obtain zero birefringence, the UV exposure time and copolymer content of photosensitive polyimides are controlled. Zero birefringence combined with photoprocessing of the material has excellent potential for applications in high-performance wavelength division multiplexing components such as polarization-independent arrayed waveguide gratings and Bragg wavelength filters. PMID- 14759059 TI - Jet-type, water-cooled heat sink that yields 255-W continuous-wave laser output at 808 nm from a 1-cm laser diode bar. AB - A newly designed jet-type, water-cooled heat sink (the funryu heat sink, meaning fountain flow in Japanese) yielded 255-W cw laser output at 808 nm from a 1-cm bar made from InGaAsP/InGaP quantum-well active layers with a 67% fill factor [70 quantum-well laser diode (LD) array along the 1-cm bar]. A funryu heat sink measuring 1.1 mm in thickness gave the LD 0.25 degrees C/W thermal resistance, one of the lowest values achieved with a 1-cm LD bar. Over a short period of operation, the device reached a maximum cw power of 255 W. To the best of our knowledge, this is the highest power ever achieved in 808-nm LD operation. In the future, the funryu heat sink may be capable of 80-W cw operation over an extended lifetime of several thousand hours. PMID- 14759060 TI - Atomic absorbers for controlling pulse propagation in resonators. AB - We consider pulse propagation through a Fabry-Perot cavity with silver mirrors that contain macroscopic samples of resonant absorbers. We show that the pulse velocity can be tuned from subluminal to superluminal in a strongly coupled atom cavity system. We delineate the effects of the interplay of cavity and absorbers. We demonstrate the saturation effects of pulse advancement with increasing mirror thickness and atomic damping. PMID- 14759061 TI - Optical clockwork with an offset-free difference-frequency comb: accuracy of sum- and difference-frequency generation. AB - We demonstrate a simple optical clockwork mechanism based on the broadened frequency comb of a femtosecond laser and on difference-frequency generation (DFG) in a nonlinear crystal. The DFG comb possesses a vanishing carrier envelope offset frequency that permits the construction of a simple and thus potentially more stable optical clockwork. In addition it offers the possibility of extending the frequency comb into the infrared spectral region. The overall accuracy and stability of the DFG comb relative to the initial frequency comb were measured to be 6.6 x 10(-21) and 10(-18) tau(-1), respectively, where tau is the averaging time in seconds. Assuming that sum- and difference-frequency generation are independent processes, our measurements suggest a <10(-20) accuracy for them. PMID- 14759062 TI - Time-wavelength two-dimensional femtosecond fluorescence imaging. AB - We report a new femtosecond time-resolved fluorescence spectrometer that enables us to observe fluorescence intensity as a time-wavelength two-dimensional image in a single measurement. This method utilizes a time-to-space conversion technique and fluorescence sum-frequency mixing with a femtosecond gate pulse. It provides a fluorescence image covering temporal and spectral spans of approximately 2 ps and approximately 60 nm, respectively. Calibration of the time and intensity axes of the image is made by use of a long-lived dye fluorescence. The two-dimensional fluorescence image of beta-carotene obtained demonstrates the high potential of this method for quantitative studies of ultrafast excited-state dynamics. PMID- 14759063 TI - Asymmetry of the neuroendocrine system. PMID- 14759064 TI - Assault on the thyroid by xenobiotics: another attack on G-protein coupled. PMID- 14759065 TI - In vitro assay of thyroid disruptors affecting TSH-stimulated adenylate cyclase activity. AB - Several natural or synthetic chemicals have been indicated as potential thyroid disruptors. The development of in vitro assays has been recommended to comprehensively assess the potential thyroid disrupting activity of a substance or a complex mixture. In this study, 12 substances suspected for acting as thyroid disruptors were tested for their ability to inhibit TSH-stimulated cAMP production in vitro. Those substances producing an inhibition were further studied to establish the level at which they interfere with this step of thyroid cell function. Using Chinese hamster ovary cells (CHO) transfected with the recombinant human TSH receptor, a dose-dependent inhibition of TSH-stimulated adenylate cyclase activity was produced by 1,1-bis-(4-chlorphenyl)-2,2,2 trichloroethan (DDT), Aroclor 1254 and Melissa Officinalis. All three substances also inhibited the cAMP production stimulated by TSH receptor antibody. Melissa Officinalis produced a significant inhibition of TSH binding to its receptor and of antibody binding to TSH, while no significant changes were produced by Aroclor 1254 or DDT in these assays. These data suggest that principles contained in Melissa Officinalis may block the binding of TSH to its receptor by acting both on the hormone and the receptor itself, while DDT and Aroclor 1254 affect cAMP production mainly at post-receptor step. In conclusion, we have developed a set of in vitro assays that allow investigation into the effect of thyroid disruptors on the TSH-mediated activation of the cAMP cascade. These assays may be useful to identify the mechanism of action of thyroid disruptors, coming beside and supporting animal studies or epidemiological surveys. PMID- 14759066 TI - New roles for pituitary transcription factors. PMID- 14759067 TI - Human pituitary tumours express the bHLH transcription factors NeuroD1 and ASH1. AB - Among the transcription factors involved in pituitary ontogenesis and physiology, basic helix-loop-helix (bHLH) have been poorly studied. Members of bHLH family include NeuroD1 and ASH1, both involved in neuroendocrine differentiation. We evaluated their mRNA expression patterns, by semi-quantitative RT-PCR analysis (sq-RT-PCR) and/or Northern blot, in a series of 33 pituitary adenomas (PA), anterior pituitaries, and pituitary cell lines. Immunohistochemistry for NeuroD1 was also performed in 25 PA. Low levels of NeuroD1 were observed in normal pituitaries and in the somatomammotroph cell lines GH3/GH4C1, contrasting with high levels in corticotroph AtT20 cells. NeuroD1 mRNA was widely expressed in PA (82%), with measurable levels found especially in those derived from Pit-1 independent lineages, i.e. corticotroph (5/5) and clinically non-secreting (CNS) adenomas (9/11). According to sq-RT-PCR analysis, overexpression of NeuroD1 compared to normal pituitaries was frequent. Variable nuclear NeuroD1 immunopositivity was also present in about 70% of studied cases. ASH1 mRNA was widely detected in normal pituitaries, in all tumour cell lines and in most PA (84%), with measurable levels in corticotroph (5/5) and CNS (9/11) adenomas, and in a significant subset of PA derived from Pit-1 dependent lineages (9/16). We conclude that: a) NeuroD1 is differentially expressed in PA and its possible ontogenetic and/or pathogenetic implications in non-corticotroph PA are discussed; b) ASH1 is a neuroendocrine marker whose expression is largely conserved in normal and neoplastic pituitary cells. PMID- 14759068 TI - DT56a stimulates creatine kinase specific activity in vascular tissues of rats. AB - The novel natural product DT56a (Secure Pharmaceuticals, Yavne, Israel), derived from soybean, has been shown to relieve menopausal vasomotor symptoms and increase in bone mineral density with no effect on sex steroid hormone levels or endometrial thickness. In single injection, like 17beta-estradiol (E2), DT56a stimulated bone, cartilage and uterus in immature or ovariectomized female rats, by measuring the changes in the specific activity of the BB isozyme of creatine kinase (CK). When administered in multiple oral doses, DT56a stimulated skeletal tissues similarly to E2 but not uterine CK. The selective estrogen receptor modulator raloxifene blocked the stimulation of CK by either DT56a or by E2 in all tissues tested. In the present study we measured the effects of DT56a on vascular tissues i.e. aorta (Ao) and the left ventricle of the heart (Lv). Both types of animals responded to either single or multiple administration of DT56a like to E2. In the Ao from both animals and in the Lv from ovariectomized rats, raloxifene completely blocked CK activity induced by DT56a, whereas in the Lv of immature female rats the inhibition was partial. Our experimental findings suggest that DT56a acts as estrogen; it has beneficial effects not only on skeletal tissues, but also on vascular tissues, however contrary to estrogen DT56a, did not affect the uterus. These findings suggest that DT56a--which has similar beneficial effects on vascular tissues like that of E2--is probably mediated via common receptor(s). PMID- 14759069 TI - Effects of a mixture of polychlorinated biphenyls (Aroclor 1254) on the transcriptional activity of thyroid hormone receptor. AB - Polychlorinated biphenyls (PCBs) are environmental contaminants which may affect thyroid function. PCBs may reduce serum thyroid hormone (TH) concentrations by either displacing T4 from TH transport proteins or increasing its hepatic metabolism. The reduced serum T4 causes neurological and growth defects in animals exposed to PCBs during the perinatal period, which can partially be reverted by T4 administration. In addition to a hypothyroid-like syndrome, a direct action of PCBs on TH-sensitive genes has been postulated. In the present study the effects of Aroclor 1254 (ARO), a mixture of PCBs, on transcription of TH-dependent genes were investigated. A reporter plasmid containing the TH responsive element (TRE) of malic enzyme (ME) gene was used in transient transfections to assess the responsiveness to ARO. ARO (10 microM) reduced the CAT activity by about 50% and competed with T3 to reduce the induction of transcription. Cotransfection of TH receptor (TR) and a wild type TRE was required to reveal ARO inhibitiry effect, which was abolished by a mock reaction not containing TR or by a mutated TRE. ARO reduced the 125I-T3 binding to TR by 30%, but did not affect the interaction of TR with a 32P-labeled TRE in gel shift assay. ARO is likely to produce a conformational change in in vitro translated TR, leading to its increased proteolysis by trypsin. These results demonstrate that ARO interacts with TR, thereby affecting the transcription of TH-sensitive genes, and provide a molecular basis to further explain the complex effects of PCBs on TH disruption. PMID- 14759070 TI - T4 but not T3 administration is associated with increased recurrence of Graves' disease after successful medical therapy. AB - TSH has been incriminated in Graves' disease for increasing the production of antibodies against TSH receptor (TRAb). It has been, therefore, suggested that T4 administration after successful antithyroid drug (ATD) treatment may indirectly decrease the production of TRAb and, therefore, the frequency of recurrence of hyperthyroidism. To study the role of T4 and T3 on the recurrence rate of Graves' disease 108 patients with Graves' disease (22 males, age: 49.8 +/- 14.3 yr, mean +/- SD, and 86 females, age: 41.7 +/- 12 yr) were followed-up for 24 months after successful treatment with ATD (carbimazole). During the follow-up period, patients daily received either 100 microg T4 or 25 microg T3 or placebo after random and double-blinded assignment into three groups. They were evaluated trimonthly up to 12 months and at 24 months. Plasma TRAb levels were measured at the beginning and at 12 months. At 12 months of the follow-up period, 14 out of 33 (42.4%), 6 out of 38 (15.8%), and 9 out of 37 (24.3%) patients receiving T4, T3 and placebo, respectively, recurred. Recurrence rate of T4-treated patients was statistically higher than that of the T3-treated patients or controls (p < 0.05). At the beginning of the follow-up period patients who were going to recur had significantly higher TRAb levels and goiter weight than patients who were not (p < 0.05). At 24 months of the follow-up period, from the patients who did not drop out of the study, none out of 11 (0%), 2 out of 19 (10.5%) and 1 out of 12 (8.3%) receiving T4, T3 and placebo, respectively, recurred. We conclude that T4 administration after successful ATD treatment of Graves' disease is associated with increased recurrence of hyperthyroidism as compared to the T3 or placebo administration. High TRAb levels and goiter weight at the end of ATD treatment may hint at recurrence. PMID- 14759071 TI - Lack of association between UCP2 gene polymorphisms and obesity phenotype in Italian Caucasians. AB - The importance of the genetic component on adipose tissue accumulation has been clearly demonstrated. Among the candidate genes investigated, there are those that regulate thermogenesis and, thus, can affect energy expenditure. The uncoupling proteins (UCPs) are a family of proteins that uncouple respiration leading to generation of heat and increased energy expenditure. Contradictory data indicate that allelic variants in their coding genes might be associated with obesity. In this study we evaluated the role of two allelic variants of the UCP2 gene in obesity and the association with its sub-phenotypic characteristics. To this aim, 360 morbidly obese patients [age: 45 +/- 15 yr, body mass index (BMI): 46 +/- 7 kg/m2] and 103 normal weight subjects (BMI < 24 kg/m2) were genotyped for the 45 bais-pair (bp) insertion/deletion (I/D) in the 3' untraslated region of exon 8 of the UCP2 gene while the presence of an Ala/Val substitution at codon 55 (Ala55Val) of the same gene was studied in 104 obese and 50 lean subjects. Patients also underwent a study protocol including measurements of BMI, waist-to-hip ratio (WHR), resting energy expenditure (REE), energy intake, fat mass (FM) and free fat mass (FFM), total cholesterol (TCH), high density lipoprotein (HDL) cholesterol, triacylglyceroles (TG), leptin levels, basal glucose, immunoreactive insulin (IRI), glycated haemoglobin (HbA1c), insulin sensitivity and thyroid hormones. No significant association between the two polymorphisms studied and the clinical, metabolic and anthropometric parameters characteristic of the obese phenotype was found. These results, in accordance with similar findings previously obtained in other ethnic groups, suggest that these two UCP2 allelic variants may not have a direct role in the pathogenesis and development of obesity. PMID- 14759072 TI - Normal values for thyroid ultrasonography, goiter prevalence and urinary iodine concentration in schoolchildren of the Veneto Region, Italy. AB - Goiter prevalence in school-age children and median urinary iodine concentration (UIC) are the main indicators of iodine deficiency in a population. In areas of mild iodine deficiency, where goiters are small, ultrasound is preferable to physical examination to estimate goiter prevalence. The World Health Organization (WHO) has adopted thyroid volume ultrasonography results from a survey of European schoolchildren as an international reference, but these values have recently been questioned. The aims of the study were: a) to determine regional normal echographic reference values of thyroid volume in children aged between 11 and 14 yr in the Veneto Region, in North-East Italy; b) to determine goiter prevalence by physical and ultrasonographic examination; c) to determine UIC in this section of the population. A cross-sectional study was carried out on 1730 schoolchildren, aged between 11 and 14, living in towns in low-lying areas, in the valleys of the pre-Alps and in the mountains between 600 and 1200 m. Thyroid volume was evaluated by inspection and palpation using the WHO criteria. In 560 children thyroid volume was determined by ultrasound. UIC was measured in 1368 children. On physical examination a grade I goiter was found in 7.5% of children. No goiter grade II or grade III was found. The regional thyroid volume reference values by ultrasonography were similar, or slightly lower (5-20%), to the corresponding WHO reference values. Mean UIC was 148 +/- 110 microg/l, with no difference between lowlands and uplands; UIC values less than 100 microg/l were found in about 30-35% of the children. UIC was higher in children using iodized salt than in non-users. No correlation was found between thyroid volume by ultrasonography and UIC. Thyroid volume was found to be bigger in upland children than in those in low-lying areas, probably because of low iodine intake in people living in the mountains in previous generations. This data show that Veneto is not a iodine-deficient area, with no presence of endemic goiter. However, the great number of children with a UIC of less than 100 microg/l also suggests the use of iodized salt in the Veneto Region. PMID- 14759073 TI - TSH receptor and Gs(alpha) genetic analysis in children with Down's syndrome and subclinical hypothyroidism. AB - The prevalence of thyroid diseases in children with Down's syndrome (DS) is about 3%. The most frequently observed condition is autoimmune subclinical hypothyroidism (SH). Autoimmune SH must be distinguished from defects in the biological activity of the TSH molecule or from the rare inherited condition of thyroid resistance to TSH. To investigate this last aspect we studied 12 patients with DS that had moderately elevated TSH with normal free thyroid hormones without signs of autoimmunity. For the genetic analysis the genomic DNA was extracted from peripheral lymphocytes. All the exons of the TSH receptor (TSHr) and Gs(alpha) genes were sequenced. The genetic analysis of the TSHr gene revealed the presence of four polymorphic variants. In two patients there was an allelic variant in the exon 1 (Pro52Thr--in one patient in the heterozygous state and in the other as a homozygous substitution). In one patient there was an allelic variant in the exon 1 (Asp36His) in the heterozygous state. In 11 patients there was a silent polymorphism in the exon 7 at nucleotide 561. All patients were homozygous for a silent polymorphism in the exon 9 at nucleotide 855. No inactivating mutations of TSHr or Gs(alpha) genes were identified in the 12 patients. In conclusion, our results seem to exclude the role of TSHr or Gs(alpha) gene mutations in the pathogenesis of the non-autoimmune SH observed in some children with DS. PMID- 14759074 TI - Determination of oxidative protein and lipid damage in adult hypopituitary patients with GH deficiency. AB - The aim of this study is to determine oxidative protein and lipid damage in adult hypopituitary GH-deficient patients. Eighteen hypopituitary GH-deficient- otherwise healthy-adults on conventional replacement therapy other than GH (9 male, 9 female, age 41.8 +/- 16.4 yr) and 18 healthy subjects (6 male, 12 female, age 40.3 +/- 16.2 yr) participated in the study. Plasma products of oxidative protein damage [protein carbonyl (PCO) and nitrotyrozine (NT)], plasma oxidized LDL (oxLDL), plasma product of oxidative lipid damage [lipid hydroperoxide (LHP)] and antioxidant status of the plasma [total thiol (T-SH)] were measured. Body fat percentage, total and LDL-cholesterol concentrations were significantly higher in the hypopituitary group. Plasma PCO, NT, LHP and T-SH concentrations did not differ significantly between patients and controls. OxLDL concentration was significantly higher in the hypopituitary patients (62.4 +/- 17.8 vs 43.1 +/- 11.3 U/l, p = 0.001). In the patients, oxLDL correlated significantly with the duration of hypopituitarism (r = 0.6323, p = 0.01). In the controls, oxLDL correlated significantly with blood pressure, total and VLDL-cholesterol concentrations. Increased oxLDL concentration may indicate increased oxidative stress within the vascular compartment and may contribute to the proatherogenic state in GH-deficient hypopituitary patients independent from conventional risk factors. PMID- 14759075 TI - Localization and expression of thyroid hormone receptors normal and neoplastic human thyroid. AB - The aim of this study was to investigate the regional expression of thyroid hormone nuclear receptor forms (TR(alpha) and TR(beta)) and isoform (TR(alpha1) and TR(beta2)) mRNAs in normal and neoplastic (benignant and malignant) human thyroid tissue. Tumor specimens from patients with thyroid carcinomas (papillary: 5 cases; follicular: 5 cases; anaplastic: 2 cases), thyroid follicular adenomas (7 cases) and tissue from normal thyroid glands (12 cases) were analyzed by in situ hybridization and semiquantitative RT-PCR for the expression of TR(alpha1) and beta, as well as for the isoform alpha2 that does not bind the hormone. In normal tissues, TR(alpha2) was expressed at lower levels compared to TR(alpha1) (alpha1/alpha2 = 4.3). In papillary and follicular carcinomas, the expression of TR(alpha1) and TR(beta) did not change as compared with normal thyroid tissue and adenomas (0.87 +/- 0.15 SD vs 0.89 +/- 0.17 densitometric units, DU, and 0.15 +/- 0.02 vs 0.14 +/- 0.03 DU, respectively). However, the expression of TR(alpha2) was significantly higher in differentiated carcinomas compared to normal thyroid tissue and adenomas (0.47 +/- 0.05 vs 0.20 +/- 0.05 DU, p < 0.05) with alpha1/alpha2 = 1.4. In anaplastic carcinoma all TRs were absent. We concluded that both normal and pathological thyroid tissues, with the exception of anaplastic carcinoma, express all TRs in thyreocites and that differentiated thyroid carcinomas are associated in enhancing the expression of TR(alpha2) mRNA. PMID- 14759076 TI - Serum levels of sex hormones and corticosterone throughout 4- and 5-day estrous cycles in Fischer 344 rats and their simulation in ovariectomized females. AB - Among inbred strains of rats, the Fischer 344 (F344) is commonly used in immunological and behavioral studies. However, little is known about patterns of sex hormones and corticosterone (CORT) secretion throughout the estrous cycle in this strain, which is characterized by a marked CORT response to stress and variable length of cycles. In the current study, using radioimmunoassays, we assessed serum levels of progesterone, estradiol, LH, testosterone, prolactin and CORT, at 1-h intervals throughout the estrous cycle in F344 female rats with 4- and 5-day cycles, as well as in males. Vaginal smears were obtained from 268 females for 15 consecutive days to determine individual length of the estrous cycle and the exact estrous phase upon blood withdrawal, which was conducted once in each rat on the 12th day of smearing. The results indicated that both 4- and 5 day cyclers have two distinct and marked surges of progesterone, one on proestrus day and the other on diestrous-1 day. Testosterone levels in 5-day cyclers peaked on diestrus-3, one day earlier than in 4-day cyclers. Daily peak levels of CORT gradually increased from estrus day to proestrous day, whereas daily nadir levels of CORT remained unchanged. To simulate the natural kinetics of specific sex hormones in ovariectomized females, different doses of estradiol, progesterone, testosterone, prolactin or CORT were injected s.c. or i.p., or 90-day sustained release pellets containing different doses of estradiol or progesterone were implanted. The findings indicated dose- and time-dependent effects, suggesting regimens for modeling the estrous cycle or replacement therapy. PMID- 14759077 TI - Three-week thyroxine withdrawal thyroglobulin stimulation screening test to detect low-risk residual/recurrent well-differentiated thyroid carcinoma. AB - Measurement of serum TSH-stimulated thyroglobulin (Tg) is recognized as a sensitive method for detecting residual/recurrent well-differentiated thyroid carcinoma (WDTC) in patients previously treated by surgery and radioactive iodine (RAI) ablation therapy. WDTC patients who have an undetectable serum Tg on thyroid hormone therapy (THT) in the absence of Tg-antibody interference are considered to be at low risk for residual/recurrent disease. Traditional management has been to withdraw T4 for 4-6 weeks or T3 for 2 weeks to stimulate endogenous TSH. However, this prolonged THT withdrawal induces hypothyroidism and its concomitant morbidity. In the present study, we assess the efficacy of shortening the time of T4 withdrawal to only 3 weeks for detecting residual/recurrent WDTC as a sufficient serum TSH stimulus for obtaining a positive serum Tg result without a routine diagnostic whole body scan (WBS). Additionally, we have evaluated the impact of such a T4 withdrawal interval on quality of life and loss of employment time. A total of 181 patients with WDTC selected for study had previously been treated with a bilateral surgical thyroidectomy followed by RAI ablation therapy (average post-surgery to follow-up interval of 10.8 yr). All of the cohort had an undetectable (< 1 microg/l) serum Tg on THT without Tg-antibody interference. Serum TSH and Tg were measured before and after cessation of T4 therapy for 3 weeks. A serum Tg > or = 2 microg/l was considered positive for residual/recurrent disease. A quality of life questionnaire [Short-Form 36 (SF-36)] was administered before withdrawal, at peak TSH and after resumption of therapy. From the completed SF-36 questionnaires, the overall degree of functional impairment was not severe and did not result in loss of employment time. Moreover, this protocol identified three possible responses to the 3-week T4 withdrawal interval as follows: a) serum Tg undetectable with TSH > or = 25 mIU/l (approximately 75% of total cohort); b) serum Tg > or = 2 microg/l (approximately 10% of total cohort) which will require further investigation and treatment for residual/recurrent disease; c) undetectable serum Tg with inadequate TSH rise (approximately 15% of total cohort), which will require TSH stimulation by either longer T4 withdrawal or recombinant human TSH to exclude residual disease. We conclude that a stimulated serum Tg test performed 3 weeks after T4 withdrawal is a simple and cost-effective first-line screening test with minimal morbidity which is sufficient to evaluate low-risk WDTC patients for recurrent/residual carcinoma. PMID- 14759078 TI - A rare case of orbital involvement in Riedel's thyroiditis. AB - We report a case of Riedel's thyroiditis in a 59-yr-old Caucasian female. She presented hypothyroidism and a stony hard, painful goiter. Due to fever, a high sedimentation rate and a high C-reactive protein (CRP), as well as high levels of anti-thyroid peroxidase antibodies (anti-TPOab), differential diagnostic considerations included acute and subacute thyroiditis as well as Hashimoto's thyroiditis and thyroid malignancy. At the same time the patient had clinical and radiological features of bilateral orbital pseudotumor with lacrimal gland involvement. During L-thyroxine therapy orbital symptoms and signs improved and thyroid size decreased. This case report serves as a reminder of differential diagnostic considerations in the etiology of goiter as well as ophthalmopathy. Although extremely rare, orbital sclerosing fibrosis can be seen in conjunction with Riedel's thyroiditis as part of multifocal fibrosclerosis. PMID- 14759079 TI - Necrobiosis lipoidica diabeticorum: response to pentoxiphylline. AB - Necrobiosis lipoidica diabeticorum (NLD) is a condition that can be physically and psychologically distressing. Angiopathy leading to thrombosis and occlusion of the cutaneous vessels has been implicated in its etiology. Pentoxiphylline is a hemorrheological agent that improves blood flow and decreases red cell and platelet aggregation. Based on these data, aim of our study was to report clinical course of a 20-yr-old diabetic woman with NLD during therapy with Pentoxiphylline 400 mg 3 times daily. After 1 month of therapy, the lesions stopped enlarging. After 3 months, the lesions showed initial signs of healing. At 6-month follow-up, there was near resolution of the lesions. The patient continued therapy and remained in remission at 2-yr follow-up. This improvement relieved psychological stress on the patient. No side effects of treatment were reported. In conclusion, patients with NLD may benefit from treatment with pentoxiphylline. We recommend therapy with 400 mg 3 times daily. The drug should be continued for at least 6 months. PMID- 14759080 TI - Antagonizing the cannabinoid receptor type 1: a dual way to fight obesity. PMID- 14759081 TI - Cushing's syndrome in pregnancy. PMID- 14759082 TI - Endocrinology and Art. Louise ou la Servante Bretonne. PMID- 14759083 TI - L-arginine ameliorates the abnormal sympathetic response of the dysfunctional human coronary microvasculature. AB - A nitric oxide (NO)-related defect may contribute to abnormal coronary sympathetic responses that can cause ischemia in patients with endothelial dysfunction. Because L-arginine, the NO synthase (NOS) precursor, augments NO bioactivity, we hypothesized that L-arginine would improve dysfunctional coronary sympathetic responses. Eleven patients with atherosclerosis or its risk factors were challenged with the cold pressor test, a specific provocative test of cardiac sympathetic activity, after 3 separate and sequential intracoronary infusions, as follows: 1) Normal saline; 2) L-NMMA, a competitive inhibitor of NOS; and 3) L-arginine. Study patients exhibited abnormal microvascular responses with coronary vascular resistance (CVR) increasing by 22.3 +/- 9.7% (mean +/- 1 SEM), p < 0.01. In addition, the change in coronary blood flow (CBF) did not correlate with the change in rate pressure product (RPP), r = -0.29, p = NS, suggesting an uncoupling of CBF from cardiac work. In the presence of L-NMMA, the CVR response, 10.3 +/- 9.8%, did not differ from the baseline response, and there was no relationship between the changes in CBF and RPP, r = 0.13, p = NS. In contrast, L-arginine ameliorated the CVR response, -3.2 +/- 3.1%, p < 0.05 vs baseline response, and restored the normal correlation between the changes in CBF and RPP, r = 0.74, p < 0.01. L-arginine not only improved abnormal microvascular responses to sympathetic activation, but it also restored the coupling that normally exists between coronary blood flow and cardiac work. L-arginine warrants further investigation as a therapy for coronary artery disease. PMID- 14759084 TI - Coronary blood flow in evolving myocardial infarction preceded by preinfarction angina: a critical reevaluation of preconditioning effects in clinical cases. AB - This study was undertaken to reevaluate the protective effects of preinfarction (pre-MI) angina in acute MI. The mechanisms involved in the apparent protective effects of pre-MI angina have been presumed to be preconditioning effects as defined by experimental studies. The phenomenon, has not, however, been observed in diabetic and/or elderly patients or in those treated by primary percutaneous coronary intervention (PCI). A total of 202 patients with anterior wall MI without a history of MI who underwent primary PCI with coronary balloon dilation and stenting (rate: 50%) <6 hours after onset were studied. Patients included 59 with pre-MI angina (group 1) and 143 without pre-MI angina (group 2). The infarct related coronary artery was patent on admission in 46% of group 1 and 31% of group 2 (p=0.045). Thrombolysis in Myocardial Infarction (TIMI) 1-2 flow was significantly more frequent in group 1 (29%) than in group 2 (11%, p=0.005) on admission. Among risk factors, clinical background, coronary anatomy, and clinical outcome, the only significant predictor of pre-MI angina was a patent infarct-related coronary artery on admission (odds ratio: 2.39, p = 0.015). There was no significant difference in left ventricular ejection fraction, peak creatine kinase, or the incidences of heart failure and in-hospital/follow-up deaths between these groups. In conclusion, the findings suggest that the protective effects reported in MI with pre-MI angina treated by thrombolysis are due to more fragile thrombotic occlusion, which can be more easily recanalized by thrombolysis, whereas the beneficial effects are not evident in those treated by primary PCI. PMID- 14759085 TI - Warfarin and aspirin versus aspirin alone in patients with acute myocardial infarction: a pilot study. AB - The benefits of anticoagulant therapy and antiplatelet agents in secondary prevention of myocardial infarction (MI) are well known. Administration of combined warfarin and aspirin (ASA) has not been well studied. The objective of this study was to compare the effect of coadministration of warfarin and ASA with administration of ASA alone on outcome of patients after MI. One hundred forty age- and sex-matched survivors of MI were randomized to receive either 100 mg/day ASA plus enough warfarin to reach a target: international normalized ratio of 2.5 (range: 2-3) (group A, n = 70), or only 100 mg/day ASA (group B, n = 70). The patients were examined for several variables including development of hemorrhage, reinfarction, and rehospitalization for 1 year post MI. Of the variables studied, minor hemorrhagic episodes were observed significantly (p = 0.002) more in group A than in group B patients. Rehospitalization and reinfarction rates, although occurring with lower frequencies in group A than in group B, did not reach the statistical significance level. In postmyocardial infarction patients, warfarin plus ASA did not provide a clinical benefit beyond that achievable with aspirin monotherapy, and for the observed markedly higher incidence of minor hemorrhage in combination therapy, antiplatelet therapy alone seems to be a more reasonable approach. PMID- 14759086 TI - Evaluation of a Doppler-derived index combining systolic and diastolic left ventricular function in acute myocardial infarction. AB - Assessment of left ventricular (LV) function is crucial in the immediate postinfarction period. The authors evaluated the clinical applicability of the Doppler-derived myocardial performance index (MPI, defined as the sum of isovolumic contraction and relaxation times divided by LV ejection time) in patients with acute myocardial infarction (AMI) as to whether this index reflects the severity of LV dysfunction in this subgroup of patients. Post-AMI patients (n = 33) were compared with age- and sex-matched healthy subjects (n = 35). Within 24 hours of the AMI and 1 month thereafter, patients underwent 2D and Doppler echocardiography. Patients were divided into group A (Killip Class I, n = 22) and group B (Killip Class II-III, n = 11). The authors measured the LV ejection fraction (EF), diastolic indices (transmitral E and A waves, E/A ratio, deceleration time [DT], isovolumic contraction time [IVCT], isovolumic relaxation time [IVRT], MPI, LV end-systolic and end-diastolic volume indices [ESVi and EDVi] and wall motion score index [WMSi]). One-year mortality was also assessed. There was no significant difference concerning E and A waves, E/A ratio, and IVRT between the 2 groups. There were highly statistical differences at day 1 for EF (59.3 +/- 6.7% vs 36.8 +/- 4.5%, p<0.0001), DT (0.160 +/- 0.030 sec vs 0.127 +/- 0.022, p<0.005), MPI (0.344 +/- 0.084 vs 0.686 +/- 0.120, p<0.0001), ESVi (28.4 +/- 3.9 mL/m2 vs 46.2 +/- 8.4, p<0.001), and WMSi (1.58 +/- 0.06 vs 1.88 +/- 0.35, p=0.05), which persisted after 1 month. One-year mortality was significantly (0 vs 27.3%, p<0.01) lower in group A patients. This study shows that the MPI, reliably indicated LV dysfunction post-AMI, significantly correlated with clinically determined functional class, and possibly has some prognostic implication. PMID- 14759087 TI - Are coronary artery anomalies an accelerating factor for coronary atherosclerosis development? AB - The relevance of benign congenital coronary anomalies (CAAs) in the atherosclerotic process is still confused despite the number of single reports of coronary artery disease in CAAs. The present study is aimed at assessing the role of CAAs on the progression of coronary artery disease (CAD). A review of the last 15,000 coronary angiographies was performed to select patients with CAAs, and they were divided into 2 groups on the basis of the presence (group I) or the absence (group II) of CAD. Clinical and instrumental records of the selected patients were reviewed and the numbers of cardiovascular events for each group (acute myocardial infarct, unstable angina, silent ischemia, bypass or percutaneous transluminal coronary angioplasty (PTCA) interventions, and cardiovascular death) were recorded from the date of diagnosis to July 2002. Group I (22 patients, mean age 64.1 +/- 9.1 years, F/M = 10/12) and group II (17 patients, mean age 66.5 +/- 10.6 years, F/M = 7/10) were similar for age and ejection fraction values. The presence of risk factors was statistically higher in group I. The number of patients with cardiovascular events was significantly higher in group I: 50% (11 patients) in group I vs 12% (2 patients) in group II, p<0.05). Repeated coronary angiography in 8/11 patients of group I and in the 2 patients of group II confirmed that the causes of the events were precedent atherosclerotic lesions in 7 patients and newly developed lesions in 3. At a mean follow-up of 60.4 +/- 12.3 months, mean actuarial survival was lower in group I than in group II (74.8% vs 100%, p=0.045), whereas mean event-free survival was 41.7% in group I and 88.7% in group II (p=0.02). Benign CAAs do not seem per se to be an accelerating factor for coronary atherosclerosis development in patients with no or few classical risk factors. PMID- 14759088 TI - Dynamics of cutaneous laser Doppler flux with concentration of moving blood cells and blood cell velocity in legs with venous ulcers and in healthy legs. AB - Laser Doppler flux (LDF) is a product of the concentration of moving blood cells (CMBC) and the blood cell velocity (BCV). In an attempt to obtain more information about the cutaneous microcirculation in legs with venous ulcers and in healthy legs, the dynamics of the curves of the LDF, the CMBC, and the BCV were analyzed in 8 patients with venous leg ulcers and in 10 subjects with healthy legs. The curves of the CMBC and of the BCV were found to be in opposite phases both in the ulcerous legs and in the healthy legs. The maximal amplitude of the curves of the LDF and of the CMBC was greater in the legs with ulcers than in the healthy legs (p = 0.021 and p = 0.0085, respectively). The finding that the curves of the CMBC and the BCV were in opposite phase can be interpreted to reflect the capillary blood flow by fits and starts both in legs with venous ulcers and in healthy legs. The greater amplitude of the LDF and the CMBC in legs with venous ulcers reflects the blood flow in anatomically altered capillaries in those legs. It is useful to record the curves of the CMBC and the BCV together with the LDF because this gives additional information about the microcirculation of the skin. PMID- 14759089 TI - Prevalence of peripheral arterial disease and related risk factors in an urban Mexican population. AB - Peripheral arterial disease (PAD) is a growing and often underdiagnosed health problem that predicts cardiovascular events and mortality. Estimating its prevalence in the general population is a major issue for assessing health needs and planning health services. The aim of this study was to determine the prevalence of PAD and its risk factors in an urban Mexican population. A random sample of 400 adult subjects was selected from a Family Medical Unit of the Mexican Institute of the Social Security. Clinical examination was performed and a questionnaire was applied to all subjects. After an overnight fast, serum glucose, triglyceride, and cholesterol concentrations were measured. Blood pressure was taken and the ankle-brachial index (ABI) was calculated by Doppler examination in both sides. PAD was diagnosed if one of the ABIs was less than 0.90. Prevalence was estimated with 95% confidence intervals (CI95%), and odds ratios (OR) with CI95% were obtained to assess association with some atherogenic risk factors in a multiple logistic regression analysis. The prevalence of PAD was 10.0% (CI95%, 7.24%-13.37%), and it was higher in men. Most subjects with PAD had no signs or symptoms, although the presence of either signs or symptoms was more frequent in subjects with PAD. The main risk factors related to PAD were serum triglycerides > or = 150 mg/dL (OR 2.25; CI95% 1.0-5.1), heavy smoking (OR 2.5; CI95% 0.9-6.7) and a history of diabetes mellitus for longer than 7 years (OR 1.9; CI95% 0.6-5.8). The prevalence of PAD is high in this Mexican urban population. Asymptomatic PAD may be highly frequent, and low-cost, noninvasive Doppler ultrasonography should be considered as an adequate screening procedure in primary care to detect individuals at high risk for major cardiovascular events. PMID- 14759090 TI - The effect of short-term treatment with simvastatin on renal function in patients with peripheral arterial disease. AB - The objective of this study was to investigate the effects of lipid-lowering treatment on renal function in patients with peripheral arterial disease (PAD). This was a retrospective study of hyperlipidemic claudicants referred to a vascular surgery and risk modification clinic. Serum creatinine and urate concentrations and the fasting lipid profile were measured pretreatment and after 3-4 months of treatment with 20 mg/day simvastatin. In 103 consecutive patients with PAD (57 men; 46 women), median age 67 years (range: 51 to 83) there was a significant decrease in serum creatinine from a mean (SD) of 87 (12) micromol/L pretreatment to 84 (12) micromol/L post-treatment (p<0.0001). This difference was more marked in the tertile of patients with the highest baseline creatinine levels. There was also a significant reduction in serum urate from 0.37 (0.07) mmol/L to 0.35 (0.07) mmol/L (p<0.0001). Both these effects were independent of the degree of total cholesterol (TC) or low-density lipoprotein (LDL) cholesterol reduction. There was a significant reduction in TC from 6.6 (1.0) to 5.2 (0.8) mmol/L and LDL cholesterol from 4.3 (1.0) to 2.8 (0.7) mmol/L; both p<0.0001. Significant improvement also occurred in the high-density lipoprotein cholesterol and triglyceride levels. Cholesterol lowering with simvastatin 20 mg/day improved indices of renal function after 3-4 months of treatment in hyperlipidemic patients with PAD. Further studies are needed to establish and define the clinical relevance of these findings, especially in patients with different degrees of renal failure. PMID- 14759091 TI - Effect of left bundle branch block on systolic and diastolic function of left ventricle in heart failure. AB - This study was designed to examine the effect of left bundle branch block (LBBB) on systolic and diastolic function of the left ventricle (LV) in patients with heart failure and in normal subjects. Thirty-six patients with heart failure and LBBB (group I), 36 patients with heart failure with normal conduction (group II), and 41 subjects with isolated LBBB (group III) were compared. Coronary angiography was performed and LV end diastolic pressure was calculated. Echocardiography was performed on all patients. LV ejection fraction and mean rate of circumferential shortening were calculated. The following Doppler parameters were evaluated: peak rapid filling velocity (E wave), peak atrial filling velocity (A wave), E- and A-wave integrals, E-wave acceleration time and deceleration time (EDT) and rates (EAR and EDR), the E/A ratio and its integral, and diastolic flow time (DT). The ejection time, isovolumetric relaxation time (IRT), and preejection period were measured using the aortic and mitral flow. LV end diastolic pressure was calculated as 28 +/- 4 mm Hg, 22 +/- 5 mm Hg, and 15 +/- 3 mm Hg in groups I, II, and III, respectively. Although the systolic function parameters in group III patients were different, the diastolic function parameters of group II were found to be quite similar to those of group III patients. Comparison of group I patients with group II patients showed that there was a similarity between LV systolic function parameters while the diastolic function parameters were different (E/A, p = 0.004; EAR, p < 0.001; EDR, p < 0.001; EDT, p < 0.001; IRT, p = 0.024; DT, p = 0.03). In conclusion, this study evaluating the effects of LBBB in normal subjects (isolated LBBB) and patients with heart failure showed that LBBB causes diastolic function impairment in normal subjects similar to those of patients with heart failure, and also increases impairment of diastolic function in patients with heart failure. PMID- 14759092 TI - Congenital familial vascular anomalies: a study of patients with an anomalous inferior vena cava, and of their first-degree relatives. AB - With magnetic resonance angiography and computed tomography, congenital anomalies of the inferior vena cava are diagnosed more frequently than they used to be. Accessory renal arteries identified by magnetic resonance angiography in a patient with an anomalous inferior vena cava indicated a combination of arterial and venous abnormalities. The study was initiated to screen consecutive patients with an anomalous inferior vena cava for concomitant abdominal and pelvic arterial abnormalities, and their first-degree relatives for congenital vascular anomalies. Magnetic resonance angiography identified in 2 of 5 patients with an anomalous inferior vena cava concomitant accessory renal arteries and in 5 of 11 first-degree relatives major abdominal vascular anomalies including accessory renal arteries, accessory renal veins, and anomalies of the hepatic artery. None of the relatives showed abnormalities of the inferior vena cava. The familial occurrence of vascular anomalies strongly suggests an underlying pathogenetic component in affected family members. In patients with a congenital anomaly of the inferior vena cava, concomitant arterial abnormalities should be considered. First-degree relatives may be at risk for congenital vascular anomalies. PMID- 14759093 TI - Electrocardiologic and echocardiographic features of patients exposed to scorpion bite. AB - The purpose of this study is to examine clinical progress and hemodynamic and electrocardiologic features (QT depression and heart rate variability [HRV]) of patients exposed to a scorpion bite. Seventeen patients bitten by scorpions, and, as a control group, 15 healthy subjects were included in the study. Standard electrocardiograph (ECG) records, 24-hour Holter-ECG, and Doppler echocardiographic examinations were performed. Holter ECG indicated sinus tachycardia, sinus bradycardia, paroxysmal supraventricular tachycardia, atrial fibrillation, first-degree and second-degree atrioventricular block not requiring treatment, early atrial beats, and early ventricular beats in the patients at frequencies of 82%, 12%, 35%, 12%, 8%, 70%, and 47%, respectively. HRV parameters that reflected parasympathetic activity (SD 35+/-13-43+/-16, RMS-SD: 20+/-9-30+/ 12, high frequency: 7.8+/-2-4.3+/-3, p<0.05) were significantly lower (p<0.05). Low frequency, which especially showed sympathetic activity (LF: 11+/-13-11+/-23, p>0.05), was similar in both groups. In addition, the LF/HF ratio, which reflected sympathovagal balance, was significantly increased in the patient group (1.5+/-1-3.0+/-2, p=0.005). Corrected QT and QT dispersion values were not significantly different with respect to the control (p>0.05). In the patient group compared to the control, a significant decrease was determined in the proportion of mitral E velocity to mitral A velocity (mEv/mAv), diastolic filling period (DFP), and left ventricular ejection fraction (LVEF), while a significant increase was noticed in pulmonary artery pressure (PAP) (mEv/mAv: 0.9+/-0.4-1.7+/ 0.6, DFP: 362+/-8.5-425+/-89, LVEF: 53.1+/-6.7-68.6+/-5.8, PAP: 38.1+/-13-27.2+/ 6, p<0.05). Scorpion bite leads to serious cardiovascular disorders, associated with decreased HRV, decreased systolic and diastolic functions, increased arrhythmic events, and hemodynamic disturbance with sympathetic and parasympathetic balance disturbance. PMID- 14759094 TI - Fever and inflammatory response as principal manifestation of chronic aortic type A dissection--a case report. AB - Aortic dissection usually presents with sudden onset of severe pain. Unfortunately, in more than half of these patients, the diagnosis is missed on admission. If the patient survives, the aortic dissection becomes chronic, whereas the patient may have few or minor symptoms. A rare manifestation may be subfebrile temperatures and inflammatory response. The authors describe a patient in whom aortic dissection resulted in night sweats, weight loss, and subfebrile temperatures. PMID- 14759095 TI - New-onset congestive heart failure secondary to a "huge" right atrial mass--a case report. AB - Transesophageal echocardiography is a useful adjunct to other diagnostic modalities in uncovering the etiology of congestive heart failure. The authors describe the case of a 75-year-old woman with a 4-week history of progressive congestive heart failure, in whom transesophageal echocardiography played a critical role in the diagnosis of a right atrial mass, accounting for this patient's constellation of symptoms. PMID- 14759096 TI - Capecitabine-induced coronary vasospasm--a case report. AB - The authors report the case of a 70-year-old man with metastatic colon cancer and no known history of coronary disease or major risk factors who developed coronary vasospasm after the initiation of capecitabine (Xeloda). Although coronary vasospasm has been associated with another older fluoropyrimidine compound, 5 fluorouracil, this is the first reported occurrence with this relatively newer cancer drug. PMID- 14759097 TI - Pulmonary endarteritis and subsequent embolization to the lung as a complication of a patent ductus arteriosus--a case report. AB - The authors describe a case of pulmonary endarteritis and subsequent embolization to the lungs as a complication of a patent ductus arteriosus (PDA). Although 2 dimensional echocardiography has been shown to be of great value in the diagnosis of patients with infective endocarditis, echocardiographic detection of vegetation within the pulmonary artery and subsequent embolization to the lung is extremely rare and, to our knowledge, has been previously reported only in a few cases. In brief, our case not only shows the importance of echocardiography in making this rare diagnosis but also emphasizes the role of echocardiography as an effective means of following up such a case. PMID- 14759098 TI - Origin of all major coronary arteries from left sinus of Valsalva as a common coronary trunk: single coronary artery--a case report. AB - Coronary anomalies are divergent and can occur in up to 1% to 2% of patients. The most common of these anomalies is separate ostia of the left anterior descending and left circumflex arteries, followed by origin of the circumflex coronary artery from the right coronary artery and the left coronary artery from the right sinus of Valsalva, either as a separate ostium or as a part of single coronary artery. Anomalous origin of right coronary artery from the left sinus of Valsalva with a separate ostium or from the left main coronary artery is very rare. These coronary anomalies may be incidentally diagnosed on routine angiography or may present with myocardial ischemia, infarction, or sudden death. A case is described in which all 3 coronary arteries were originating from the left sinus of Valsalva as a common trunk (single coronary artery), which trifurcated to left anterior descending, left circumflex, and right coronary artery. PMID- 14759099 TI - Left atrial thrombosis in achalasia--a case report. AB - An 81-year-old woman came to the clinic with aphasia and gait disturbance. A transthoracic echocardiogram showed left atrial (LA) thrombus at the posterobasal site, which was compressed by a large mass with high echo intensity. A chest computed tomography scan revealed dilatation of the esophagus, compressing the LA from the posterior side. An endoscopy showed a bolus and much foulness in the distended esophagus, which were immediately eliminated by an endoscopic procedure. The LA thrombus, as shown by transesophageal echocardiography, spontaneously resolved without any neurologic signs. The authors assumed that the dilatated esophagus compressed the LA, and this may have produced the abnormal flow dynamics in the LA. The injury of endocardial surface produced by the distension or turbulent flow, in addition to dehydration, may have augmented a coagulability in the LA. PMID- 14759100 TI - Quadfurcation of the left main coronary artery: double ramus medianus coronary system. PMID- 14759101 TI - Extra-oral endosseous craniofacial implants: current status and future developments. AB - The published experience of extra-oral endosseous craniofacial implants (EOECIs) is reviewed. The definition of osseointegration, concept of success, the relative merit of one- or two-stage implant placement, EOECI design and control of peri abutment skin infection are discussed. A plea is made for more consistent and objective reading of clinical experience of this technique. PMID- 14759102 TI - Juvenile mandibular chronic osteomyelitis: a distinct clinical entity. AB - Sclerosing osteomyelitis of the mandible is an uncommon disease of unknown aetiology. A series of eight female children (6 to 12 years old) with a distinct mandibular inflammatory disease were studied. Each presented with pain and a recurrent soft tissue swelling overlying a predominantly unilateral mandibular enlargement. On imaging, this deformity demonstrated a mixture of patchy sclerosis and radiolucency. A raised erythrocyte sedimentation rate was the only consistent serological finding. Treatment varied from symptomatic control with non-steroidal anti-inflammatory medication, to surgical management that included decortication and contouring and, in one case, resection with reconstruction. A potential protocol for treatment of this disease is given. The early age of onset of the disease process and the uniformity of the features distinguish this condition from other groups of disorders that, previously, have been collectively designated as chronic diffuse sclerosing osteomyelitis. It is proposed that this inflammatory disease of mandibular bone, in the paediatric patient, should be regarded as a separate clinical entity: 'juvenile mandibular chronic osteomyelitis'. PMID- 14759103 TI - Computed tomographic findings of the fractured mandibular condyle after open reduction. AB - The purpose of this study was to evaluate the long-term radiological results obtained with open reduction and fixation of unilateral mandibular condyle fractures in 10 patients. CT images taken at the end of the follow-up period (average of 22 months, range 7 to 33 months), were traced and digitized, and the position and morphology of the fractured condylar process was statistically compared with those of the contralateral non-fractured condylar process in the coronal, transverse and sagittal planes. Little difference was observed in the position or morphology of the condylar process in the operated and non-fractured joints. This study shows that it is possible to anatomically reduce fractured condyles, and thereby to avoid postoperative disadvantageous joint changes. PMID- 14759104 TI - Long-term results following miniplate eminoplasty for the treatment of recurrent dislocation and habitual luxation of the temporomandibular joint. AB - Twenty patients with recurrent mandibular dislocation or habitual luxation of the temporomandibular joint who underwent miniplate eminoplasty between 1994 and 1999 are presented. The follow-up period ranged from 2 to 7 years. Clinical examination showed no evidence of recurrent dislocation. No significant reduction of normal mouth opening was found one year postoperatively. The pain level, self estimated by the patients on a scale from 0 to 10, decreased significantly from 4.1 preoperatively to 1.2 one year postoperatively. In seven patients fractures of the miniplates occurred 3 to 7 years after eminioplasty (two bilateral fractures, five unilateral fractures). Due to the high incidence of plate fractures miniplate eminoplasty cannot be recommended as the surgical treatment of choice for recurrent dislocation or habitual luxation of the mandible, although the recurrence rate is low and pain is reduced considerably. PMID- 14759105 TI - Modified temporalis anchorage in craniomandibular reankylosis. AB - This study evaluates the long-term outcomes and clinical results of costochondral graft and temporalis muscle flap interpositioning with submandibular anchorage in the management of TMJ re-ankylosis. Thirty-one patients, 9 children and 22 adults, with recurrence of ankylosis after gap arthroplasty, with a mouth opening less than 5 mm were evaluated. The management protocol consisted of resection of the ankylotic mass through an Al-Kayat and Bramley incision; contralateral coronoidectomy in unilateral cases; replacement of the condyle in children by means of a costochondral graft through Risdon's approach and interpositional temporalis muscle flap and submandibular anchorage of the temporalis flap in children and adults. Regular clinical and radiological follow up was done for 6 years during which the average mouth opening of 38 mm was maintained, with good occlusion and proper function. The temporalis muscle flap was seen to be an ideal interpositional material due to its close proximity to the site, good vascular supply, ease of access to the condyle area and minimal risk of nerve damage. Submandibular anchorage of the broad temporalis muscle flap prevents reankylosis by inhibiting flap contraction, and decreases need for rigorous physiotherapy. PMID- 14759106 TI - Fear of recurrence and possible cases of anxiety and depression in orofacial cancer patients. AB - Recurrence risk in orofacial cancer patients is relatively high in comparison to other cancers. Little information exists on whether patients are unduly concerned over the possibility of recurrence and the association of this concern with psychological morbidity. Aim of study was to assess fear of recurrence and psychological morbidity in orofacial cancer patients. Two samples of patients with orofacial cancer were followed prospectively. First sample (n = 87) were consecutively drawn 3 months and 7 months following initial treatment. Second sample (n = 100) were cross-sectionally drawn from out-patient list and followed 2 years later. A single question item was employed to assess fear of recurrence used in previous work. Hospital Anxiety and Depression Scale was adopted as the measure of psychological morbidity. A prospective survey design was employed. Over 80% of patients expressed concern over the possibility of recurrence at 3 months post treatment. This level reduced to 72% at 7 months (P=0.06). Approximately two-thirds of patients sampled cross-sectionally were concerned at both assessment occasions. Psychological morbidity was greatest at 3 months post treatment (possible cases: anxiety 37% and depression 28%). Women were more likely to report anxiety (at possible case level) than men 3 months following treatment (P<0.05). Patients aged 65 or more years were less concerned about recurrence. This effect was significant on both occasions that sample two patients were assessed (P's<0.002). Very few patients who expressed no concern about recurrence, at 3 months, were found to report anxiety or depression sufficient to be recognized as a possible case (3 and 0% respectively). The positive association between psychological morbidity and fears of recurrence was significant at the majority of data collection points, with the exception that depression was more independent of these concerns. PMID- 14759107 TI - Influence of bone invasion and extent of mandibular resection on local control of cancers of the oral cavity and oropharynx. AB - The aim of this paper was to evaluate the influence of bone invasion on treatment outcome among patients with cancers of the oral cavity and oropharynx and to determine whether or not outcome was influenced by the extent of mandibular resection. A review of 127 prospectively documented patients who were treated with marginal or segmental resection for oral (n = 110) and oropharyngeal (n = 17) cancers was undertaken. There were 97 males and 30 females with a median age of 61 years. Clinical T stages were: T1 17 patients, T2 33, T3 22, T4 55. Median followup was 4 years. A total of 94 patients underwent marginal resections and 33 underwent segmental resections. Histological bone invasion was present in 17 patients (16%) in the marginal resection group and 21 patients (64%) in the segmental group (P<0.05). Soft tissue surgical margins were positive in 11 patients (12%) in the marginal group and in seven patients (21%) in the segmental group (P=not significant). Local control did not correlate significantly with T stage, the extent of mandibular resection or the presence of histological bone invasion, but was significantly influenced by positive soft tissue margins (P<0.01). Among patients with bone invasion, the local control rate was higher following segmental resection when compared to marginal resections (87% vs 75%) but this was not statistically significant. Survival was significantly influenced by positive soft tissue margins but not bone invasion or the type of resection. We conclude that bone invasion alone did not predict for local control or survival rates among patients with oral and oropharyngeal cancers. Involved soft tissue margins were highly predictive of local recurrence and decreased survival. Conservative resection of the mandible is safe as long as marginal mandibulectomy does not lead to compromise of soft tissue margins. Segmental resection should be reserved for patients extensive bone invasion or those with limited invasion in a thin atrophic mandible. PMID- 14759108 TI - A randomized prospective clinical trial on the effectiveness of three treatment modalities for patients with lower denture problems. A 10 year follow-up study on patient satisfaction. AB - Treatment of lower denture complaints of patients with an edentulous mandible with a height of at least 15 mm can consist of meticulous construction of a new set of dentures (CD), construction of a new set of dentures following preprosthetic surgery to enlarge the denture-bearing area (PPS), or construction of an implant-retained mandibular overdenture (IRO). The aim of this prospective randomized controlled clinical trial was to evaluate the satisfaction of the above mentioned treatment modalities in resolving lower denture related complaints. Ninety edentulous patients (Cawood class IV and V, mean mandibular height 20.7 +/- 2.7 mm) were randomly assigned to one of these treatment modalities. The main outcome parameters were denture satisfaction and chewing ability, which were assessed using validated self-administered questionnaires focusing on denture-related complaints and problems with chewing different types of food. These parameters were measured before treatment, and 1, 5 and 10 years after treatment. Differences among the three groups were tested by applying a one way analysis of variance and a Tukey's test for multiple comparison. At the 1 year evaluation, significantly better scores were observed in the two surgical groups (IRO, PPS) than in the CD group (P<0.05). At 5 year evaluation the 'complaints of the lower denture' showed a significantly better score in the IRO group when compared to the PPS and CD groups (P<0.05). No significant differences were observed between the PPS and CD group. At 10 year evaluation, the intention to treat analysis revealed no significant differences between the three groups, while a per protocol analysis showed that the IRO group was the most satisfied. From this study it is concluded that both on the short and long term denture satisfaction appears most favourable in the IRO group when compared to the PPS and CD groups. Implant-retained overdentures are, therefore, a favourable treatment modality for edentulous patients with lower denture problems. PMID- 14759109 TI - Tranexamic acid mouthwash--a prospective randomized study of a 2-day regimen vs 5 day regimen to prevent postoperative bleeding in anticoagulated patients requiring dental extractions. AB - This prospective randomized study analyses the use of a prescribed 4.8% tranexamic acid post-operative mouthwash over 2 days vs 5 days to prevent bleeding in patients taking warfarin who require dental extractions. Eighty-five patients therapeutically anticoagulated with warfarin for various conditions, ranging in age from 21 to 86 years and requiring dental extractions, were randomly divided into two groups. Group A postoperatively received a 4.8% tranexamic acid mouthwash to be used over a 2-day period. Group B received the same mouthwash and instructions postoperatively, to be taken over 5 days. All procedures were performed on an ambulatory basis under local anaesthetic by the same surgeon. Patients were reviewed 1, 3, and 7 days postoperatively to assess bleeding. Eighty-two of the 85 patients encountered no postoperative problems. Two patients in group A and one in group B had minor postoperative bleeds that required minor ambulatory intervention to control. This study shows that a 2-day postoperative course of a 4.8% tranexamic acid mouthwash is as equally effective as a 5-day course in controlling haemostasis post-dental extractions in patient's anticoagulated with warfarin. PMID- 14759110 TI - Prefabrication of vascularized bone flap induced by recombinant human bone morphogenetic protein 2 (rhBMP-2). AB - An experimental model for the prefabrication of a vascularized bone flap was developed in this study. To form vascularized bone in the desired configuration and to increase the survival rate of the grafted bone, a muscle vascularized pedicle (MVP) was transformed into vascularized bone by the inducer recombinant human bone morphogenetic protein 2 (rhBMP-2). The muscle flap (8 x 8 mm) raised on saphenous vessels in the rat thigh was sandwiched between same-size collagen (Terudermis) sheets in the presence or absence of impregnated 25 microg of rhBMP 2 for the experimental group and the control group, respectively. The flaps were harvested 1, 2 and 3 weeks postoperatively. Bone transformation was detected by gross examination, radiology, and histologic testing. No evidence of muscle tissue transformation was found in control flaps, whereas all of the experimental flaps produced new bone. Saphenous vessels were observed to supply the new bone upon harvesting, and the newly formed vascularized bone showed good configuration with shape of the Terudermis sheet. This study indicates that this model of effective bone reconstruction could be potentially applied in a therapeutic setting. PMID- 14759111 TI - Reconstruction of maxillectomy defect by transport distraction osteogenesis. AB - The study aimed to explore the feasibility of posterior maxillectomy reconstruction by transport distraction in a primate model. In each of 14 male adult rhesus monkeys, posterior partial maxillectomy was performed on one side of maxilla to create a posterior maxillary deflect. Immediately after the maxillectomy, a dentoalveolar segment anterior to the defect was osteotomized as transport segment and a custom-made transport distractor was fixed on the residual maxilla. After a latency period of 5 days, the distractor was activated 1 mm daily to move the transport segment backward to the defect. This process lasted about 2 weeks. The transport segment was allowed to consolidate and the animals were sacrificed at different defined intervals. Transport distraction was successful in six animals. Three other cases were completed with minor wound dehiscence and one had a small oro-antral fistula with subsequent maxillary sinusitis. New bone bridging the distraction gap was confirmed by radiography and histology in the animals completing distraction. Reconstruction of posterior maxillectomy defect is proven feasible by transport distraction osteogenesis. PMID- 14759112 TI - Development of a mechanical testing system for a mandibular distraction wound. AB - The purpose of this study was to develop a mechanical testing system to estimate stiffness of an experimental porcine mandibular distraction osteogenesis (DO) wound. The system was designed to function without changing the morphology of the healing mandible. A customized jig was designed to allow cantilever-bending tests of the Yucatan mini-pig hemi-mandible. Experimental and control hemi-mandibles were placed in the jig and the proximal segment was secured. A material testing unit applied progressively increasing downward force on the pre-molar occlusal surface. The maximum force applied was 0.030 kN. The stiffness value for each hemi-mandible was represented by the slope of the plot of force (kN) vs displacement (mm). Radiographs were taken before and after mechanical testing to demonstrate any gross morphologic changes or identifiable fractures across the distraction wound. A total of 24 mini-pigs underwent DO of the right mandible with 0-day latency and distraction rates of 1, 2, and 4 mm per day resulting in a 12 mm gap. At the completion of 0, 8, 16, and 24 days of neutral fixation, two animals for each of three different distraction rates were sacrificed for mechanical testing. Stiffness of control hemi-mandibles ranged between 0.018 and 0.317 kN/mm (median 0.063; mean 0.099 +/- 0.080). Stiffness of experimental hemi mandibles ranged between 0 and 0.025 kN/mm (median 0.004; mean 0.005). The subset that was tested at the end of neutral fixation had stiffness between 0.005 and 0.025 (median 0.011; mean 0.012 +/- 0.011). No morphologic changes were evident on the X-rays after testing. The results indicate that the cantilever-bending model is useful for testing stiffness of an experimental mandibular DO wound without destroying its morphology. PMID- 14759113 TI - Acceleration of callus maturation using rhOP-1 in mandibular distraction osteogenesis in a rat model. AB - The aim of this study was to assess if the application of rhOP-1 induces accelerated consolidation of the callus in mandibular distraction osteogenesis. In seven adult Wistar rats a bilateral osteotomy of the horizontal ramus of the mandible was performed in the molar region and a custom designed distractor was mounted to the mandible. With a rate of 0.7 mm per day the device was activated bilaterally after the seventh postoperative day. After seven days of distraction two times 50 microg rhOP-1 were injected on two subsequent days directly into the callus. The contralateral side received an injection of placebo solution. The animals were killed four weeks after the end of distraction. A three-point bending test revealed a significantly higher strength of the distracted mandible in the rhOP-1 side (66.3 N vs. 30.4 N, P=0.034, paired t-test). Undecalcified histological sections were examined using microradiography and fluorescence microscopy after sequential intravital polychromic labelling. A continuous bony bridging was seen in all rhOP-1 sites and in none of the control sites. The data indicate that rhOP-1 may be an option to accelerate callus maturation in mandibular distraction osteogenesis. PMID- 14759114 TI - Expression of inducible nitric oxide synthase in cutaneous adnexal tumours of the head and neck. AB - Adnexal tumours are a rare heterogeneous group of neoplasms, most commonly found in the head and neck region. Although most are benign, malignant adnexal tumours often behave aggressively, resulting in early metastasis. There is increasing interest in the role that nitric oxide (NO) plays in the behaviour of many cancers. It is thought that NO, produced by the enzyme inducible NO synthase (iNOS), facilitates tumour growth and dissemination. iNOS has been studied in the common skin cancers, where its expression correlates with tumour behaviour, but it has not been previously investigated in cutaneous adnexal tumours. An immunhistochemical study was performed using a monoclonal antibody to iNOS in 37 cases of adnexal tumours (19 benign, 18 malignant). iNOS expression was weakly expressed by basal keratinocytes of adjacent skin in all cases and it was variably expressed in the tumours. Malignant tumours had significantly increased iNOS expression when compared to both adjacent skin (P<0.001) and the benign tumour group (P<0.001). No significant difference was found between iNOS expression in benign tumour and adjacent skin (P=0.5). The role of iNOS in this rare group of tumours and the possibility of pharmacologically inhibiting it in the clinical setting warrants further investigation. PMID- 14759115 TI - Human papillomavirus, p53 and cyclin D1 expression in oropharyngeal carcinoma. AB - Forty-two specimens from oropharyngeal (tonsil and base of tongue) squamous cell carcinoma patients (SCC) were studied for presence of HPV 16 by in situ hybridization and by immunohistochemistry for p53 and Cyclin D1 protein overexpression. Thirty-one per cent of cases were HPV-16 positive, which correlates with the prevalence reported worldwide. 74% of cases showed p53 protein overexpression and 79% showed Cyclin D1 overexpression. There was no correlation between HPV status and either p53 or Cyclin D1 overexpression (P>0.05). These three variables also did not correlate with factors such as grade of the tumour, stage of the disease or lymph nodal metastasis at presentation. PMID- 14759116 TI - Subcutaneous fat layer in different donor regions used for harvesting microvascular soft tissue flaps in slender and adipose patients. AB - The aim of this study is to provide valid data concerning the thickness of the subcutaneous fat layer in seven donor regions which are most commonly used for soft tissue replacement in the head and neck region. Furthermore the study focuses on differences in thickness of this layer depending on the nutritional status and sex. In 100 volunteers the thickness of the subcutaneous fat layer was evaluated using ultrasound in seven potential donor regions: radial and ulnar forearm, lateral upper arm, scapular and parascapular region, abdominal/supra umbilical region and the lateral calf. Volunteers were divided into 4 groups: male and female, slender (body mass index [BMI] < 25) and adipose (BMI > 30). Data obtained was compared and differences were proven to be statistically significant (Student's t-test, P<0.05). The largest thickness was found in the supra-umbilical region followed by the lateral upper arm whereas the ulnar forearm region had the smallest thickness. Depending on the BMI the supra umbilical and the lateral upper arm regions showed the most wide variation. Depending on sex the most noticeable variation in thickness of the subcutaneous layer was seen in the lateral calf region with a significant larger tissue layer in females. PMID- 14759117 TI - Pattern of third molar impaction in a Singapore Chinese population: a retrospective radiographic survey. AB - One thousand orthopantomograms (OPGs) of patients 20-40 years old were examined. Where impacted third molars were present, the angle and depth of impaction were recorded. Results were analysed using the Pearson chi2 test. 68.6% of OPGs showed at least one impacted third molar. The frequency was three-fold higher in the mandible (1024/1079=90%) than in the maxilla (306/1077=28%), with a significantly higher frequency (P<0.05) in females (56%) than males (44%). The mesioangular impaction was the most common, and 80% of all impacted third molars were partially buried in bone. Of the 429 bilateral occurrence of impacted third molars, 423 were in the mandible. It was concluded that the frequency of impacted third molars in the Singapore Chinese population studied was generally two to three times that reported in races of the Caucasian stock. There was also double the frequency of impacted third molars when compared to a previous study in a Chinese population published in 1932 with females being more frequently affected than males. PMID- 14759118 TI - Maxillary distraction using a trans-sinusal distractor: technical note. AB - In this pilot study, the principle of distraction osteogenesis was used to advance the midface of a boxer dog. A modified high Le Fort I-type osteotomy was performed. Following a latency period of 5 days the maxilla was distracted 14 mm in 14 consecutive days at a rate of 1 mm per day. Ten weeks after the completion of the distraction, multiple biopsies were taken across the distraction gap. Histological observation showed bone deposition in the osteotomy sites. Soft and hard tissue formation resulted in complete healing across the distraction gap. The maxillary sinus was used to accommodate the distraction device. Superimposition of the standardized lateral cephalograms taken at the end of distraction and 14 months after the removal of the distractors showed no sign of relapse in the achieved sagittal advancement of the maxilla. This small, intraoral trans-sinusal placed distractor has a completely new conceptual design, and may be helpful in distraction of maxilla in children and adults with midfacial hypoplasia. PMID- 14759119 TI - Aspergillosis of the temporomandibular joint following irradiation of the parotid region: a case report. AB - We report a case of aspergillosis in the right temporomandibular joint (TMJ) with a history of parotid carcinoma and post-irradiation otitis. Previous treatment attempts with surgery and antibiotics were unsuccessful. Radical debridement of the glenoid fossae, supplemented with amphotericin B and adjunct hyperbaric oxygen (HBO) therapy, was provided to resolve the symptoms. This case report highlights the need to be aware of the possibility of invasive mycosis in immunocompromised patients. PMID- 14759120 TI - Pathologic and imaging findings of an oncocytoma in the deep lobe of the left parotid gland. AB - Oncocytoma is a rare salivary gland tumour consisting of oncocytes with many hyperplastic mitochondria. It usually occurs in the parotid gland. Because the features of oncocytoma resemble those of other benign and low-grade-malignant salivary gland tumours, clinical diagnosis is often challenging. This report presents the pathologic and imaging findings of an oncocytoma arising in the deep lobe of the left parotid gland in a 66-year-old man. Oncocytoma was diagnosed on the basis of histological, magnetic resonance imaging, and scintigraphic findings. The tumour showed accumulation of technetium-99m pertechnetate and decreased signal intensity on both T1- and T2-weighted magnetic resonance images. PMID- 14759121 TI - A basal cell adenoma of the sublingual gland. AB - To the knowledge of the authors, only one case of a basal cell adenoma of the sublingual gland has ever been reported. We report a second case of a basal cell adenoma arising from the sublingual gland in this paper. PMID- 14759122 TI - Variations in the course of the hypoglossal nerve: a case report and literature review. AB - The extracranial hypoglossal nerve has a well described course as it traverses the neck, and is frequently identified during neck dissection, carotid endarterectomy and other procedures involving the deep spaces of the neck. A case is presented which demonstrates aberrancy of its anatomic position. Variations of its course are well documented, however none have demonstrated a course lateral to the internal jugular vein. This serves to alert the surgeon to be mindful of such atypical presentations as the consequences of iatrogenic injury to such structures may be significant. PMID- 14759123 TI - Successful management of severe facial pain in patients with diffuse sclerosing osteomyelitis (DSO) of the mandible using disodium clodronate. PMID- 14759124 TI - Applications of the ninhydrin reaction for analysis of amino acids, peptides, and proteins to agricultural and biomedical sciences. AB - The reaction of ninhydrin with primary amino groups to form the purple dye now called Ruhemann's purple (RP) was discovered by Siegfried Ruhemann in 1910. In addition, imines such as pipecolic acid and proline, the guanidino group of arginine, the amide groups of asparagine, the indole ring of tryptophan, the sulfhydryl group of cysteine, amino groups of cytosine and guanine, and cyanide ions also react with ninhydrin to form various chromophores of analytical interest. Since its discovery, extensive efforts have been made to apply manual and automated ninhydrin reactions as well as ninhydrin spray reagents to the detection, isolation, and analysis of numerous compounds of interest across a broad spectrum of disciplines. These include agricultural, biochemical, clinical, environmental, food, forensic, histochemical, microbiological, medical, nutritional, plant, and protein sciences. This reaction is unique among chromogenic reactions in that at pH 5.5 it results in the formation of the same soluble chromophore by all primary amines which react, be they amines, amino acids, peptides, proteins, and even ammonia. Because the chromophore is not chemically bound to the protein or other insoluble material, it is not lost when the insoluble substrate is removed by centrifugation or filtration after the reaction is completed. The visible color of the chromophore is distinctive and is generally not affected by the yellow colors present in many food, plant, and tissue extracts. Adaptations of the classical ninhydrin reaction to specialized needs in analytical chemistry and biochemistry include the use of acid, alkaline, and fluorogenic ninhydrin reagents. To cross-fertilize information among several disciplines wherein an interest in the ninhydrin reaction has developed, and to enhance its utility, this review attempts to integrate and correlate the widely scattered literature on ninhydrin reactions of a variety of structurally different compounds. Specifically covered are the following aspects: historical perspective, chemistry and mechanisms, applications, and research needs. A better understanding of these multifaceted ninhydrin reactions provide a scientific basis for further improvements of this important analytical technique. PMID- 14759125 TI - Discrimination of recombinant and pituitary-derived bovine and porcine growth hormones by peptide mass mapping. AB - Somatotropins, which are used in cattle for growth and lactating performances, are difficult to reliably detect because no direct method exists. Reversed-phase high-performance liquid chromatography (RP-HLC) coupled to electrospray ionization quadrupole mass spectrometry (ESI/MS) has been developed to separate and characterize the N-terminal peptides resulting from tryptic cleavage of natural and recombinant growth hormones from different species (bovine, porcine, and human) and suppliers. Conditions for tryptic digestion were optimized. This technique was found to be optimal to cleave efficiently the N-terminal peptide of the proteins without releasing too much noise from the matrix. Characterization of the peptides through ESI(+)-MS allowed natural and recombinant growth hormones from bovine and porcine species with N-terminal amino acid sequences differing from one amino acid residue to be discriminated. However, the studied human growth hormones had similar primary sequences that did not permit any discrimination between recombinant and natural forms, thus confirming the known identity of these hormones. Protein digestions with pepsin and chymotrypsin were also compared but were not conclusive due to the too small N-terminal peptides released after proteolysis. PMID- 14759126 TI - On-line multisensor monitoring of yogurt and filmjolk fermentations on production scale. AB - Near-infrared (NIR) spectrometry and electronic nose (EN) data were used for on line monitoring of yogurt and filmjolk (a Swedish yogurt-like sour milk) fermentations under industrial conditions. The NIR and EN signals were selected by evaluation of principal component analysis loading vectors and further analyzed by studying the variability of the selected principal components. First principal components for the NIR and the EN signals were used for on-line generation of a process trajectory plot visualizing the actual state of fermentation. The NIR signals were also used to set up empirical partial least squares (PLS) models for prediction of the cultures' pH and titratable acidity (expressed as Thorner degrees, degrees T). By using five or six PLS factors the models yielded acceptable predictions that could be further improved by increasing the number of reliable and precise calibration data. The presented results demonstrate that the fusion of the NIR and EN signals has a potential for rapid on-line monitoring and assessment of process quality of yogurt fermentation. PMID- 14759127 TI - Potential of SPME-GC and chemometrics to detect adulteration of soft fruit purees. AB - The potential of combining solid-phase microextraction (SPME) with gas chromatography and chemometric data analysis to differentiate between pure strawberry samples (Fragraria ananassa) and strawberry samples adulterated with 10, 40, and 70% (v/v) apple puree was investigated. The method involved the extraction of aroma volatiles from the headspace of the puree samples using a SPME fiber followed by GC analysis with flame ionization detection. The principal component analysis (PCA) data matrix consisted of the relative percent peak areas of 37 compounds deemed to be significant in the differentiation of the samples on the basis of adulteration. The PCA results clearly showed that differentiation of the adulterated and unadulterated samples was possible, particularly at the higher levels of adulteration. Partial least-squares regression (PLSR) using a dummy set of Y variables (set to 0 for unadulterated and 1 for adulterated samples) resulted in clear discrimination between unadulterated purees and those containing 40 and 70% (v/v) apple. PLSR using a second set of Y variables, consisting of the actual level of adulteration, enabled quantification of apple puree with a standard error of prediction of 11.6%, implying a minimum detectable level of 25% (v/v) apple. GC-MS analysis enabled identification of the compounds with the greatest influence on sample differentiation. These compounds were identified as hexanoic acid, 2-hexenal, and alpha-farnesene, all of which are key aroma compounds in apples. PMID- 14759128 TI - Screening crucifer seeds as sources of specific intact glucosinolates using ion pair high-performance liquid chromatography negative ion electrospray mass spectrometry. AB - Seeds, of either commercial crucifer crops or some wild and weed relatives, were screened for intact glucosinolates using a previously developed ion-pair LC-MS method. This method, in contrast to GC-MS techniques, ensures the accurate measurement of all classes of glucosinolates. Many crucifer seeds contained very high concentrations of glucosinolates with low concentrations of additional pigments and secondary metabolites. The other common seed metabolites were cinnamoylcholine esters, for example, sinapine. Glucosinolates derived from homologues of l-methionine were characteristic of Brassica and related crucifer species. In addition, significant concentrations of 4-hydroxy-3 indolylmethylglucosinolate were found in the majority of Brassica species. Wild and weed species often had relatively simple glucosinolate profiles: either a single glucosinolate or a predominant glucosinolate together with trace amounts of others. Species identified with seed glucosinolate profiles suitable for purification included various Alyssum, Erysimum, and Iberis species for 3 methythiopropyl-glucosinolate and 3-methylsulfinylpropyl-glucosinolate and various Alyssum, Erysimum, and Lepidium species with very high concentrations of C4-C6 aliphatic glucosinolates. Seeds of Arabis, Barbarea, Lepidium, Moringa, and Sinapis species were good sources of aromatic glucosinolates, and Azima tetracantha was a good source for N-methoxy-3-indolylmethyl-glucosinolate. MS data are reported for all of the intact glucosinolates detected from the screening process. PMID- 14759129 TI - Total antioxidant capacity of arginine-conjugated linoleic acid (CLA) complex. AB - Conjugated linoleic acids (CLA) have shown diverse biological activities, including anti-carcinogenic, anti-atherosclerotic, anti-adipogenic, and anti diabetogenic effects. Recent reports also showed that CLA has free radical scavenging capacity, which may give health benefits to human beings. However, the application of CLA as a bioactive ingredient has been limited due to its insolubility in water. To overcome this problem, we investigated antioxidant activities of arginine (Arg)-CLA, a water-soluble CLA salt, using both 2,2' azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and 2,2'-diphenyl-1 picrylhydrazyl (DPPH) radical scavenging assays. CLA, Arg, and Arg-CLA all exerted radical scavenging activities in a dose-dependent manner in both assays. Arg-CLA at 20 mM scavenged 89 and 55% of ABTS and DPPH radicals in 3 h, respectively, whereas CLA alone quenched only 48 and 26% of them under the same conditions. The antioxidant activity of the Arg-CLA complex was found to be synergistic in ABTS assay and comparable to that of vitamin E in DPPH assay. PMID- 14759130 TI - Constituents of the leaves of Peucedanum japonicum Thunb. and their biological activity. AB - In the course of our study on the isolation and structure determination of constituents in tropical plants, we focused on Peucedanum japonicum Thunb., belonging to the family Umbelliferae. In this study, a new C(13) norisoprenoid glucoside, (3S)-O-beta-d-glucopyranosyl-6-[3-oxo-(2S)-butenylidenyl]-1,1,5 trimethylcyclohexan-(5R)-ol (1), and two new phenylpropanoid glucosides, 3-(2-O beta-d-glucopyranosyl-4-hydroxyphenyl)propanoic acid (3) and methyl 3-(2-O-beta-d glucopyranosyl-4-hydroxyphenyl)propanoate (4), were isolated from the n-butanol soluble fraction of this plant's leaves, together with five known compounds. The structures of these compounds were determined on the basis of spectroscopic evidence. In addition, all isolated compounds were examined for scavenging activity against 1,1-diphenyl-2-picrylhydrazyl radical and inhibitory activity against mushroom tyrosinase. These results suggested that 2-(4-hydroxy-3 methoxyphenyl)propane-1,3-diol (7) and 3-O-beta-d-glucopyranosyl-2-(4-hydroxy-3 methoxyphenyl)propanol (8) showed an appreciable activity in both assay systems. PMID- 14759131 TI - Polyozellin isolated from Polyozellus multiplex induces phase 2 enzymes in mouse hepatoma cells and differentiation in human myeloid leukaemic cell lines. AB - Induction of cellular phase 2 detoxifying enzymes is associated with cancer preventive potential. Quinone reductase (QR) has been used as a prototype for anticarcinogenic phase 2 enzymes because of its widespread distribution in mammalian systems, large amplitude of inducer response, and ease of measurement in murine hepatoma cells. Methanol extract of Polyozellus multiplex, which shows a strong QR induction potential, was subjected to bioassay-guided fractionation, and polyozellin (PM1) appeared to be a major active component. In in vitro cultured cells (hepa1c1c7 and BPRc1 cells), polyozellin enhanced QR, glutathione S-transferase (GST) activities, and glutathione (GSH) content in a dose-dependent manner. The compound also significantly promoted differentiation of HL-60 human promyelocytic emia cells. In conclusion, polyozellin deserves further in vivo study to evaluate its potential as a cancer preventive agent. PMID- 14759132 TI - Relationship of light quantity and anthocyanin production in Pennisetum setaceum Cvs. rubrum and red riding hood. AB - Pennisetum setaceum cvs. Rubrum and Red Riding Hood are purple-pigmented ornamental grasses when grown in high-light environments. In low-light environments, foliage appears light purple or green, and as a result, aesthetic appeal is reduced. The impact of light on anthocyanin pigmentation was compared for P. setaceum Rubrum foliage and flowers and Red Riding Hood foliage grown under different light intensities and light sources. Light environments included UV supplemental light in the greenhouse, high-pressure sodium supplemental light in the greenhouse, cool-white fluorescent light in a growth chamber, and full sun outside. Anthocyanins in two cultivars of P. setaceum were analyzed by HPLC and characterized by (1)H and (13)C NMR spectral experiments. Two anthocyanins, cyanidin 3-glucoside and cyanidin 3-rutinoside, were identified in the leaves and flowers of both cultivars and quantified by HPLC analysis. The major anthocyanin in both cultivars was cyanidin 3-glucoside and had highest concentration (0.199% fresh weight) in Rubrum leaves grown under fluorescent lights in the growth chamber with a photoperiod of 24 h and a daily light integral (DLI) of 13.3 mol m(-)(2) day(-)(1) and in Rubrum and Red Riding Hood leaves and flowers (0.097 and 0.12% fresh weight) from plants grown outside in full sun with a photoperiod ranging from 15 to 13.5 h and DLI of 42 mol m(-)(2) day(-)(1). The minor anthocyanin, cyanidin 3-rutinoside, had the highest quantity in plants grown in low-light-intensity greenhouse environments with a photoperiod ranging from 15 to 13.5 h and DLI of 2.3-7.0 mol m(-)(2) day(-)(1). The functional significance of anthocyanins in P. setaceum Rubrum is discussed. PMID- 14759133 TI - Growth inhibition of prostate cancer cells by epigallocatechin gallate in the presence of Cu2+. AB - Green tea is an effective chemopreventive agent to human prostate cancer adenoma (PCA). Epigallocatechin gallate (EGCG) inhibited the growth of PCA cells and induced apoptosis. Cu(2+) is a trace element necessary to our health. Many studies proved that bioactivity of EGCG is altered in the presence of Cu(2+). We investigated the effects of EGCG on PCA cells in the presence of Cu(2+). Also, we explored potential mechanisms via measurement of the relative chemiluminescence of growth medium for PCA cells. Chemiluminscence can be an indication of free radicals. Our test results showed that the addition of EGCG and Cu(2+) to the growth medium decreased the relative viability of androgen-sensitive and androgen insensitive human prostate cancer cells. However, the effects of EGCG on PCA cells depended on (1) the relative concentrations of added EGCG and Cu(2+) and (2) their order of addition. Our results indicated that few free radicals may be generated in vitro. If so, free radicals generated intracellularly may be a major factor behind apoptosis and growth inhibition observed in the PCA cells. Thus, EGCG might exert its effects intracellularly. PMID- 14759135 TI - Quantitative expression analysis of GH3, a gene induced by plant growth regulator herbicides in soybean. AB - Symptoms resembling off-target plant growth regulator (PGR) herbicide injury are frequently found in soybean fields, but the causal agent is often difficult to identify. The expression of GH3, an auxin-regulated soybean gene, was quantified from soybean leaves injured by PGR herbicides using real-time RT-PCR. Expression of GH3 was analyzed to ascertain its suitability for use in a diagnostic assay to determine whether PGR herbicides are the cause of injury. GH3 was highly induced by dicamba within 3 days after treatment (DAT) and remained high at 7 DAT, but induction was much lower at 17 DAT. GH3 was also highly induced at 7 DAT by dicamba + diflufenzopyr, and to a lesser extent by the other PGR herbicides clopyralid and 2,4-D. The non-PGR herbicides glyphosate, imazethapyr, and fomesafen did not significantly induce GH3 expression above a low constitutive level. These results indicate that a diagnostic assay for PGR herbicide injury based on overexpression of auxin-responsive genes is feasible, and that GH3 is a potential candidate from which a diagnostic assay could be developed. However, time course analysis of GH3 expression indicates the assay would be effective for a limited time after exposure to the herbicide. PMID- 14759134 TI - Efficient one pot extraction and depolymerization of grape (Vitis vinifera) pomace procyanidins for the preparation of antioxidant thio-conjugates. AB - Antioxidant thio-conjugates were obtained from white grape pomace by a clean and efficient one pot extraction and depolymerization method using water and cysteamine hydrochloride. To evaluate the potential of grape pomaces of different origins as sources of proanthocyanidins and conjugates, we conducted varietal comparative studies of polyphenol content, antioxidant power, and procyanidin composition. Xarel.lo proved to be the richest source of polyphenols. The total conversion into the conjugates was as high as 8 g/kg of Xarel.lo grape pomace, with a 50-fold excess of cysteamine, and 3 g/kg, with a 5-fold excess of cysteamine. After purification by preparative cation exchange and reversed phase high-performance liquid chromatography, 17 g of 63% pure 4beta-(2 aminoethylthio)epicatechin (acetate salt) was obtained from 35 kg of moist pomace. The procedure described here will make the antioxidant thio-derivatives efficiently available directly from raw plant byproducts such as grape pomace. PMID- 14759136 TI - Effect of cultivar and processing method on the contents of polyphenols in table olives. AB - Polyphenols were determined by HPLC in the juice and oil of packed table olives. The phenolic compositions of the two phases were very different, hydroxytyrosol and tyrosol being the main polyphenols in olive juice and tyrosol acetate, hydroxtyrosol acetate, hydroxytyrosol, tyrosol, and lignans (1-acetoxypinoresinol and pinoresinol) in oil. The type of processing had a marked influence on the concentration of polyphenols in olive juice and little on the content in oil. The analyses carried out on 48 samples showed that turning color olives in brine had the highest concentration in polyphenols ( approximately 1200 mg/kg), whereas oxidized olives had the lowest ( approximately 200 mg/kg). Among olive cultivars, Manzanilla had a higher concentration than Hojiblanca and Gordal. The type of olive presentation also influenced the concentration of polyphenols in olives, decreasing in the order plain > pitted > stuffed. The results obtained in this work indicate that table olives can be considered a good source of phenolic antioxidants, although their concentration depends on olive cultivar and processing method. PMID- 14759137 TI - Comparative study on pressure and temperature stability of 5 methyltetrahydrofolic acid in model systems and in food products. AB - A comparative study on the pressure and temperature stability of 5 methyltetrahydrofolic acid (5-CH(3)-H(4)folate) was performed in model/buffer systems and food products (i.e., orange juice, kiwi puree, carrot juice, and asparagus). Effects of pH and ascorbic acid (0.5 mg/g) on 5-CH(3)-H(4)folate stability in buffer systems were studied on a kinetic basis at different temperatures (from 65 to 160 degrees C) and different pressure/temperature combinations (from 100 to 700 MPa/from 20 to 65 degrees C). These studies showed that (i) the degradation of 5-CH(3)-H(4)folate in all model systems could be described by first-order reaction kinetics, (ii) the thermostability of 5-CH(3) H(4)folate was enhanced by increasing pH up to 7, (iii) 5-CH(3)-H(4)folate was relatively pressure stable at temperatures lower than 40 degrees C, and (iv) ascorbic acid enhanced both the thermo- and barostabilities of 5-CH(3) H(4)folate. In food products, temperature and pressure stabilities of 5-CH(3) H(4)folate were studied at different temperatures (70-120 degrees C) and different pressure/temperature combinations (from 50 to 200 MPa/25 degrees C and 500 MPa/60 degrees C). 5-CH(3)-H(4)folate in orange juice and kiwi puree was relatively temperature (up to 120 degrees C) and pressure (up to 500 MPa/60 degrees C) stable in contrast to carrot juice and asparagus. Addition of ascorbic acid (0.5 mg/g) in carrot juice resulted in a remarkable protective effect on pressure (500 MPa/60 degrees C/40 min) and temperature degradation (120 degrees C/40 min) of 5-CH(3)-H(4)folate. PMID- 14759138 TI - Gluconic acid consumption in wines by Schizosaccharomyces pombe and its effect on the concentrations of major volatile compounds and polyols. AB - Schizosaccharomyces pombe 1379 (ATCC 26760) yeast strain in wine substantially increases acetaldehyde and 1,1-diethoxyethane concentrations and to decreases levo-2,3-butanediol, glycerol, acetoin, and gluconic acid concentrations. In this study, S. pombe has been used for the first time to reduce gluconic acid in wine under aerobic conditions. Only acetaldehyde and acetoin exhibited significantly higher levels in the wines containing gluconic acid. The high in vitro specific activity of alcohol dehydrogenase observed may be directly related to the high production of acetaldehyde by the studied fission yeast. PMID- 14759139 TI - Investigations on anthocyanins in wines from Vitis vinifera cv. pinotage: factors influencing the formation of pinotin A and its correlation with wine age. AB - Pinotage red wines were found to contain a reaction product of malvidin 3 glucoside and caffeic acid, the so-called pinotin A. A total of 50 Pinotage wines from the vintages 1996-2002 were analyzed for the content of pinotin A, malvidin 3-glucoside, caffeic acid, and caftaric acid. Statistical analyses were performed to reveal variations in the content of these compounds and to determine the factors that influence pinotin A formation during wine aging. An exponential increase of the concentration of this aging product was observed with prolonged storage time. The most rapid synthesis of pinotin A was observed in 2.5-4 year old wines, although at this age malvidin 3-glucoside is already degraded to a large extent. This phenomenon is explained by the increased ratio of caffeic acid/malvidin 3-glucoside, which strongly favors the formation of pinotin A and makes side reactions less likely. Pinotin A formation proceeds as long as a certain level of malvidin 3-glucoside is maintained in the wines. In wines >5-6 years old degradation or polymerization of pinotin A finally exceeds the rate of its de novo synthesis. PMID- 14759140 TI - Two-dimensional correlation analysis of visible/near-infrared spectral intensity variations of chicken breasts with various chilled and frozen storages. AB - Generalized two-dimensional (2D) correlation analysis of visible/near-infrared (NIR) spectra was performed to characterize the spectral intensity variations of chicken muscles induced by either storage time/temperature regime or shear force values. The results showed that intensities of two visible bands at 445 and 560 nm increase with the storage temperature under identical treatment, possibly indicating a color change due to frozen storage. The 2D NIR correlation spectra indicated that all NIR bands reduce their spectral intensities, probably due to the water loss and compositional alterations during the freeze-thaw process as well as the tenderization development in muscle storage. The heterospectra correlating the spectral bands in both visible and NIR regions exhibited a strong correlation and suggested the sequential change between color and other developments in muscles. In addition, shear value-induced NIR spectral intensity variations detected significant differences in spectral features between tender and tough muscles. PMID- 14759141 TI - Free galactose concentrations in fresh and stored apples (Malus domestica) and processed apple products. AB - Gas chromatography was used to quantitate free galactose in Braeburn, Fuji, Red Delicious, and Spartan apples during cold storage, after thermal processing of apple slices and in juice produced using clarification and/or liquifaction enzymes. Spartan had significantly higher galactose levels as compared to Red Delicious apples, but changes in galactose in all varieties during 9 months of cold storage were insignificant. Blanching and canning decreased galactose levels, but doubling the thermal processing during canning increased the free galactose concentration detected in plant tissue. An enzymatic liquefaction aid used to prepare apple juice dramatically increased the free galactose content while a clarification aid caused only a slight increase due to its selective action on soluble pectin. These findings provide useful information for dietitians to base diet recommendations for galactosemic patients. PMID- 14759142 TI - Changes in the carotenoid metabolism of capsicum fruits during application of modelized slow drying process for paprika production. AB - A temperature profile simulating the traditional slow drying process of red pepper fruits, which is conducted in La Vera region (Spain) for paprika production, was developed. Carotenoid and ascorbic acid content, as well as moisture of fruits, were monitored during the slow drying process designed. Data obtained suggested that the evolution of carotenoid concentration, the main quality trait for paprika, directly depend on the physical conditions imposed. During the drying process, three different stages could be observed in relation to the carotenoids. The first stage corresponds to a physiological adaptation to the new imposed conditions that implied a decrease (ca. 20%) in the carotenoid content during the first 24 h. After that short period and during 5 days, a second stage was noticed, recovering the biosynthetic (carotenogenic) capability of the fruits, which denotes an accommodation of the fruits to the new environmental conditions. During the following 48 h (third stage) a sharp increase in the carotenoid content was observed. This last phenomenon seems to be related with an oxidative-thermal stress, which took place during the first stage, inducing a carotenogenesis similar to that occurring in over-ripening fruits. Results demonstrate that a fine control of the temperature and moisture content would help to positively modulate carotenogenesis and minimize catabolism, making it possible to adjust the drying process to the ripeness stage of fruits with the aim of improving carotenoid retention and therefore quality of the resulting product. In the case of ascorbic acid, data demonstrated that this compound is very sensitive to the drying process, with a decrease of about 76% during the first 24 h and remaining only at trace levels during the rest of the process. Therefore, no antioxidant role should be expected from ascorbic acid during the whole process and in the corresponding final product (paprika), despite that red pepper fruit is well-known to be rich on this compound. PMID- 14759143 TI - Effect of ripeness and postharvest storage on the phenolic profiles of Cherries (Prunus avium L.). AB - The phenolic compounds hydroxycinnamates, anthocyanins, flavonols, and flavan-3 ols of sweet cherry cultivars Burlat, Saco, Summit, and Van harvested in 2001 and 2002 were quantified by HPLC-DAD. Phenolics were analyzed at partially ripe and ripe stages and during storage at 15 +/- 5 degrees C (room temperature) and 1-2 degrees C (cool temperature). Neochlorogenic and p-coumaroylquinic acids were the main hydroxycinnamic acid derivatives, but chlorogenic acid was also identified in all cultivars. The 3-glucoside and 3-rutinoside of cyanidin were the major anthocyanins. Peonidin and pelargonidin 3-rutinosides were the minor anthocyanins, and peonidin 3-glucoside was also present in cvs. Burlat and Van. Epicatechin was the main monomeric flavan-3-ol with catechin present in smaller amounts in all cultivars. The flavonol rutin was also detected. Cultivar Saco contained the highest amounts of phenolics [227 mg/100 g of fresh weight (fw)] and cv. Van the lowest (124 mg/100 g of fw). Phenolic acid contents generally decreased with storage at 1-2 degrees C and increased with storage at 15 +/- 5 degrees C. Anthocyanin levels increased at both storage temperatures. In cv. Van the anthocyanins increased up to 5-fold during storage at 15 +/- 5 degrees C (from 47 to 230 mg/100 g of fw). Flavonol and flavan-3-ol contents remained quite constant. For all cultivars the levels of phenolic acids were higher in 2001 and the anthocyanin levels were higher in 2002, which suggest a significant influence of climatic conditions on these compounds. PMID- 14759144 TI - Comparative study of methods for DNA preparation from olive oil samples to identify cultivar SSR alleles in commercial oil samples: possible forensic applications. AB - Virgin olive oil is made from diverse cultivars either mixed or single. Those ensure different tastes and typicity, and these may be also enhanced by the region of production of cultivars. The different olive oil labels correspond to their chemical composition and acidity. Labels also may correspond to a protected origin indication, and thus, such oils contain a given composition in cultivars. To verify the main cultivars used at the source of an olive oil sample, our method is based on DNA technology. DNA is present in all olive oil samples and even in refined oil, but the quantity may depend on the oil processing technology and oil conservation conditions. Thus, several supports were used to retain DNA checking different techniques (silica extraction, hydroxyapatite, magnetic beads, and spun column) to prepare DNA from variable amounts of oil. At this stage, it was usable for amplification through PCR technology and especially with the magnetic beads, and further purification processes were checked. Finally, the final method used magnetic beads. DNA is released from beads in a buffer. Once purified, we showed that it did not contain compounds inhibiting PCR amplification using SSR primers. Aliquot dilution fractions of this solution were successfully routinely used through PCR with different SSR primer sets. This enables confident detection of eventual alien alleles in oil samples. First applied to virgin oil samples of known composition, either single cultivars or mixtures of them, the method was verified working on commercial virgin oil samples using bottles bought in supermarkets. Last, we defined a protocol starting from 2 x 40 mL virgin olive oil, and DNA was prepared routinely in about 5 h. It was convenient to genotype together several loci per sample to check whether alleles were in accordance with those of expected cultivars. Thus, forensic applications of our method are expected. However, the method needs further improvement to work on all oil samples. PMID- 14759145 TI - Metabolism of diethofencarb (isopropyl 3,4-diethoxyphenylcarbamate) in rats: identification of metabolites in urine. AB - Rats were orally given a diethofencarb (isopropyl 3,4-diethoxyphenylcarbamate) labeled with (14)C, at 300 mg/kg/day, for 4 consecutive days, and 11 metabolites in urine were purified by a combination of several chromatographic techniques. The chemical structures of all isolated metabolites were identified by spectroanalyses (NMR and MS). Ten of them were newly identified forms. Five of them were S-conjugates: three mercapturic acid conjugates, one S-methyl conjugate, and one SO-methyl conjugate. The others were two phenoxyacetic acids, hydroxyacetanilide, hydroxyisopropyl carbamate, and oxazolinone derivatives. From the results, the existence of the following reactions in rats can be concluded: (1) deethylation of the 4-ethoxy group; (2) conjugation of phenols with glutathione, gamma-glutamyltranspeptidation and depeptidation of the glutathione to form cysteine conjugates, and N-acetylation of the cysteine; (3) cleavage of the C-S linkage of cysteine conjugates followed by methylation; (4) oxidation of the S-methyl group; (5) cleavage of the carbamate linkage; (6) acetylation of the resultant amino group; (7) oxidation of the acetyl group; (8) oxidation of the isopropyl group; (9) cyclization of the oxidized isopropyl carbamate group; and (10) oxidation of the 4-ethoxy group. PMID- 14759146 TI - Removal of atrazine from water using covalent sequestration. AB - Monochlorotriazines including atrazine and its major metabolites, deethylatrazine and deisopropylatrazine, are susceptible to nucleophilic aromatic substitution. Competitive reactions to rank the relative reactivity of nucleophiles with atrazine reveal that constrained secondary amines are the most reactive. When the nucleophile is attached to a solid support, atrazine can be sequestered from solution. As proof of concept, polystyrene resins displaying constrained secondary amines are shown to sequester atrazine, deethylatrazine, and deisopropylatrazine from water. Sequestration can be followed spectrophotometrically or using a liquid chromatography mass spectrometry protocol. The kinetics of sequestration are similar to that of granulated charcoal. Evidence for covalent bond formation comes from control experiments with unreactive herbicides and degradation analysis of the solid support. Using both (1)H NMR spectroscopy and mass spectrometry, covalent adducts are identified in ratios close to what is calculated theoretically. This method for sequestration is effective at removing atrazine from pond water. PMID- 14759147 TI - Optimization of dynamic headspace extraction of the edible red algae Palmaria palmata and identification of the volatile components. AB - A new extraction method was applied to the volatile compounds of Palmaria palmata. Dynamic headspace was optimized according to an experimental design, and descriptive sensory analysis and intensity and similarity tests were performed for each extract to assess their respective representativeness. Results showed that extract obtained with crushed algae after a 30 min purge was the most representative. GC-MS analysis was then performed on this extract to identify the volatile components. Seven halogenated compounds, seven aldehydes, two ketones, three alcohols, and four miscellaneous compounds were identified. Among them, halogenated compounds were the most characteristic of red algae, and more particularly, iodoethane and iodopentane, which had yet been found in other seaweeds. PMID- 14759148 TI - 1. In vivo aroma release during eating of a model cheese: relationships with oral parameters. AB - This study aims to follow the kinetics of aroma compound release during model cheese consumption in order to clarify the relationships between flavor release and some oral parameters. Eight subjects participated in the study. Breathing, salivation, chewing, and swallowing were monitored during the eating process. Temporal nosespace analyses were performed using on-line atmospheric pressure ionization-mass spectrometry (API-MS) and off-line solid-phase Micro extraction gas chromatography-mass spectrometry (SPME-GC-MS). Flavor release profiles were obtained only for ethyl hexanoate, heptan-2-one, and heptan-2-ol. Among them, only the concentrations of ethyl hexanoate and heptan-2-one could be determined by API-MS. Absence of competition between the aroma compounds was checked for both techniques. In-nose maximum concentration (C(max)) varied with aroma compounds. However, C(max) was reached at the same time (T(max)) for the three compounds. Interindividual differences were observed for most of the parameters studied and for all of the aroma compounds. They were related to the interindividual differences among the oral parameters. The aroma release parameters C(max) and AUC (area under the curve) could be related to respiratory and masticatory parameters. In most cases, the same relationships were observed whatever the nature of the aroma compound. PMID- 14759149 TI - 2. In vivo nonvolatile release during eating of a model cheese: relationships with oral parameters. AB - This study deals with the release kinetics of nonvolatile compounds (NVC) (leucine, phenylalanine, glutamic acid, citric acid, lactic acid, propanoic acid, sodium, potassium, calcium, magnesium, chloride, and phosphates) during the eating of a model cheese and the relationships to some oral (salivary and masticatory) parameters. The aroma release has previously been characterized in similar conditions [Pionnier, E.; Chabanet, C.; Mioche, L.; Le Quere, J.-L.; Salles, C. J. Agric. Food Chem. 2004, 52, xxx-xxx (1)]. Saliva samples were collected from the tongues of eight assessors at different times during and after the chewing sequence. Atmospheric pressure ionization-mass spectrometry and/or high-performance liquid chromatography analyses have been performed on these samples in order to quantify the 12 NVC released in saliva. The maximum concentration (C(max)) in saliva varied significantly according to the compound. However, there was no significant effect of the compound on the time to reach maximum concentration (T(max)). Interindividual differences were observed for most of the parameters and for all of the NVC studied. The parameters extracted from the release profiles of the NVC were closely correlated. High T(max) and AUC (area under the curve) values could be related to high chewing time and low saliva flow rates, low chewing rates, low masticatory performances, and low swallowing rates. PMID- 14759150 TI - Composition of the essential oil of leaves, galls, and ripe and unripe fruits of Jordanian Pistacia palaestina Boiss. AB - Pistacia palaestina Boiss. (Pistacia terebinthus L. var. palaestina (Boiss.) Engl.) is a medicinal and foodstuff plant. The ripe fruits are used largely in the Middle East as a component of the so-called Zaatar, a mix of aromatic and food plants. Results of GC and GC-MS analyses of the essential oils of leaves, galls produced by Baizongia pistaciae (L.), and ripe and unripe fruits of Pistacia palaestinaBoiss. collected in Jordan are reported. Both qualitative and quantitative differences between different parts of the plant were observed. The oil was rich in monoterpenes, and the main constituents were alpha-pinene (63.1%) and myrcene (13.3%) in the leaves and alpha-pinene (49.4%), sabinene (22.8%), and limonene (8.1%) in the galls. (E)-Ocimene (33.8-41.3%), sabinene (20.3-24.1%), and (Z)-ocimene (3.8-13.0%) were the main ones in both unripe and ripe fruits. Sesquiterpenes have been detected in small quantities in leaves and fruits and in trace amounts in galls. PMID- 14759151 TI - Enzymatic synthesis of gamma-glutamylvaline to improve the bitter taste of valine. AB - The taste of several bitter amino acids is reduced, sourness produced, and preference increased by gamma-glutamylization. An enzymatic method for synthesizing gamma-Glu-Val involving bacterial gamma-glutamyltranspeptidase (GGT) was developed. The optimum reaction conditions for the synthesis of gamma-Glu-Val were 20 mM Gln, 300 mM Val, and 0.04 U/ml GGT, pH 10. After 3-hr incubation at 37 degrees C, 17.6 mM gamma-Glu-Val was obtained, with the yield being 88%. gamma Glu-Val was purified on a Dowex 1 x 8 column and then identified by NMR. PMID- 14759152 TI - A new insight into the formation of odor active carbonyls by thermally-induced degradation of phospholipids in self-assembly structures. AB - The role of molecular organization in heated aqueous dispersions of egg phosphatidylcholine (PC) and egg phosphatidylethanolamine (PE) was studied with respect to the formation of key odorants. Evidence was found for the crucial role of self-assembly structures adopted by phospholipid molecules on the quantitative composition of volatile constituents. The concentrations of seven aldehydes and one vinyl ketone were determined by isotope dilution assay in heated aqueous dispersions of PC and PE present in various ratios. Addition of PE to PC drastically decreased the amount of (E,E)-2,4-decadienal formed, which cannot be explained by the differences in the fatty acid composition of PC and PE. The free amino group in PE does not explain this phenomenon either, as replacing PE by phosphatidic acid distearylester also reduced the amounts of (E,E)-2,4 decadienal. We suggest that the type of self-assembly structure adopted by phospholipids in water significantly influences the reaction yields. However, the mechanisms leading to the preferred formation of phospholipid-derived odorants in a lamellar phase, as compared to the reversed hexagonal phase, remain unknown. PMID- 14759153 TI - Effect of antioxidants on oxidative stability of edible fats and oils: thermogravimetric analysis. AB - Thermogravimetric analysis was used to determine the oxidative stability of various edible oils (olive oil, milkfat) and triacylglycerides (triolein, trilinolein), while the effect of natural (alpha-tocopherol, ascorbic acid) and synthetic antioxidants (butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA), and tertiary butyl hydroquinone were evaluated by addition to trilinolein. Oil resistance to oxidation was obtained by measuring the increase in sample weight due to the uptake of molecular oxygen, the temperature at maximum sample weight, and the temperature at the onset of oxidation. When comparing sample weight increase, trilinolein proved to be oxidatively less stable than triolein, olive oil, and milk fat, while triolein was less stable than olive oil and milk fat. Olive oil showed significantly higher stability than milkfat when comparing the temperature at the onset of oxidation. When comparing effectiveness of antioxidants, a combination of 0.01% BHA and 0.01% BHT increased trilinolein stability the most. PMID- 14759154 TI - Antioxidative activities of fractions obtained from brewed coffee. AB - The antioxidative activity of column chromatographic fractions obtained from brewed coffee was investigated to find antioxidants and to assess the benefit of coffee drinking. The dichloromethane extract inhibited hexanal oxidation by 100 and 50% for 15 days and 30 days, respectively, at the level of 5 microg/mL. A GC/MS analysis of fractions, which exhibited oxidative activity, revealed the presence of antioxidative heterocyclic compounds including furans, pyrroles, and maltol. The residual aqueous solution exhibited slight antioxidative activity. The inhibitory activity (%) of the seven fractions from an aqueous solution toward malonaldehde formation from lipid oxidation ranged from 10 to 90 at a level of 300 microg/mL. The results indicate that brewed coffee contains many antioxidants and consumption of antioxidant-rich brewed coffee may inhibit diseases caused by oxidative damages. PMID- 14759155 TI - Incorporation and stabilization of omega-3 fatty acids in surimi made from cod, Gadus morhua. AB - Surimi containing omega-3 fatty acids from algal oil was prepared by the addition of oil-in-water emulsions or bulk oil. Emulsion and bulk oil were added separately to surimi to provide approximately 500 mg of omega-3 fatty acids per serving of surimi (85 g). Addition of the emulsion had no effect on surimi gel strength, whereas bulk oil decreased gel strength an average of 31%. All surimi treatments containing algal oil increased in Hunter b values due to the presence of carotenoids in the oil. Among cryoprotectants, sodium tripolyphosphate was the major surimi additive responsible for retarding the formation of lipid hydroperoxides and thiobarbituric acid reactive substances (TBARS). Lipid hydroperoxide and TBARS formation was lower in surimi containing bulk oil compared to surimi with emulsified oil. Both EDTA and lipid soluble antioxidants were able to decrease lipid oxidation in surimi fortified with omega-3 fatty acids. This suggests that surimi containing nutritionally beneficial omega-3 fatty acids could be developed with good oxidative stability and gel strength. PMID- 14759156 TI - Ferulic acid release and 4-vinylguaiacol formation during brewing and fermentation: indications for feruloyl esterase activity in Saccharomyces cerevisiae. AB - The release of ferulic acid and the subsequent thermal or enzymatic decarboxylation to 4-vinylguaiacol are inherent to the beer production process. Phenolic, medicinal, or clove-like flavors originating from 4-vinylguaiacol frequently occur in beer made with wheat or wheat malt. To evaluate the release of ferulic acid and the transformation to 4-vinylguaiacol, beer was brewed with different proportions of barley malt, wheat, and wheat malt. Ferulic acid as well as 4-vinylguaiacol levels were determined by HPLC at several stages of the beer production process. During brewing, ferulic acid was released at the initial mashing phase, whereas moderate levels of 4-vinylguaiacol were formed by wort boiling. Higher levels of the phenolic flavor compound were produced during fermentations with brewery yeast strains of the Pof(+) phenotype. In beer made with barley malt, ferulic acid was mainly released during the brewing process. Conversely, 60-90% of ferulic acid in wheat or wheat malt beer was hydrolyzed during fermentation, causing higher 4-vinylguaiacol levels in these beers. As cereal enzymes are most likely inactivated during wort boiling, the additional release of ferulic acid during fermentation suggests the activity of feruloyl esterases produced by brewer's yeast. PMID- 14759157 TI - Purification and characterization of a latent polyphenol oxidase from beet root (Beta vulgaris L.). AB - Polyphenol oxidase (PPO) has been extracted from beet root, in both soluble and membrane fractions. In both cases, the enzyme was in its latent state, and it was activated by sodium dodecyl sulfate. PPO was purified to apparent homogeneity. The soluble PPO purification was achieved by hydrophobic interaction chromatography and gel filtration chromatography, with apparent molecular mass of 55 kDa. The membrane PPO purification was achieved by anion exchange chromatography and gel filtration with apparent molecular mass of 54 kDa. A totally denaturing SDS-PAGE indicated the presence of a single polypeptide with an apparent molecular mass of 60 kDa for both fractions, with the band also revealed by Western blot. A partially denaturing SDS-PAGE stained a single active 36 kDa band for both fractions. Under native isoelectric focusing, a major acidic band of pH 5.2 was detected in both fractions. Kinetic characterization of PPO on the natural substrate l-dopa was carried out. PMID- 14759158 TI - Surface physicochemical properties of globulin-P amaranth protein. AB - Globulin-P, the polymerized 11S amaranth globulin, is composed of 280 kDa unitary molecules (UM, 23%) and aggregates larger than 500 kDa (A, 70%). Antibodies against these proteins were prepared to study their surface characteristics and to assess their homology with other storage proteins. Results showed that globulin-P unitary molecules and aggregates had similar reactive surfaces. A polypeptide of 56 kDa was found to be the most reactive to the antibodies assayed, followed by the acidic polypeptides. Such results support previous information, according to which these polypeptides appeared to be the most exposed on the molecule surface. Globulin-P fraction presented cross-reactivity with the remaining amaranth protein fractions: 11S-globulin, glutelins, and albumins. Globulin-P and 11S-globulin showed similar reactive surfaces whereas glutelin and albumins presented a lower cross-reactivity. The reactivity of the glutelin fraction depended on its sequence. Globulin-P fraction presented cross reactivity with quinoa globulins, and to a lesser extent with globulins of sunflower and rice. Moreover, the anti-Gp serum was unable to detect either conformational or sequence epitopes in globulins of soybean, wheat, buckwheat, rice, and rye. PMID- 14759159 TI - Acid-induced cold gelation of globular proteins: effects of protein aggregate characteristics and disulfide bonding on rheological properties. AB - The process of cold gelation of ovalbumin and the properties of the resulting cold-set gels were compared to those of whey protein isolate. Under the chosen heating conditions, most protein was organized in aggregates. For both protein preparations, the aggregates consisted of covalently linked monomers. Both types of protein aggregates had comparable numbers of thiol groups exposed at their surfaces but had clearly different shapes. During acid-induced gelation, the characteristic ordering caused by the repulsive character disappeared and was replaced by a random distribution. This process did not depend on aggregate characteristics and probably applies to any type of protein aggregate. Covalent bonds are the main determinants of the gel hardness. The formation of additional disulfide bonds during gelation depended on the number and accessibility of thiol groups and disulfide bonds in the molecule and was found to clearly differ between the proteins studied. However, upon blocking of the thiol groups, long fibrillar structures of ovalbumin contribute significantly to gel hardness, demonstrating the importance of aggregate shape. PMID- 14759160 TI - Degradation, under non-oxygen-mediated autooxidation, of carotenoid profile present in paprika oleoresins with lipid substrates of different fatty acid composition. AB - A heat-induced degradation was carried out over two crude paprika oleoresins diluted with high oleic or high linoleic oil. Occurrence of oxygen was avoided, and changes in individual carotenoids were followed over time. Degradation rate constants were not significantly different, so that carotenoid stability was not linked with unsaturation degree of the oily system. A parallel reaction to degradation, trans to cis carotenoid isomer conversion, was also denoted during the thermal treatment, and it initially showed a higher rate than degradation of cis-isomers. Both processes (isomerization and cis-isomers degradation) were finally compensated, and their development was also unaffected by the nature of the lipid profile. Under the reaction conditions, oleic and linoleic fatty acids showed the same reactivity and induced degradative reactions, equally affecting the carotenoid profile. An enhanced stability of carotenoid content and provitamin A value was not achieved with a decrease in the unsaturation level of the oily system. PMID- 14759161 TI - Determination of poly(ethylene glycol)-binding to browse foliage, as an assay of tannin, by near-infrared reflectance spectroscopy. AB - Nutritionists are interested in functional assays of tannins that do not require time-consuming and expensive extraction, such as the (14)C-labeled poly(ethylene glycol) (PEG)-binding (PEG-b) assay. This paper reports the application of near infrared (NIR) spectroscopy to determine the percentage of PEG binding, in place of the (14)C-labeled PEG-b assay of tannin, in Mediterranean woodland vegetation. Calibration was done with 53 samples from 14 species and was validated on 25 samples from 10 species. PEG-b ranged between 1.4 and 20.7% in the samples. The calibration obtained by using the modified partial least-squares (MPLS) method, with all wavelengths in the 1100-2500 nm range combined, and the validation were reasonably linear (R (2) = 0.96 and 0.91, respectively). The accuracies, estimated from the standard errors of cross-validation and prediction, were +/ 1.6 and +/-1.7% PEG-b, respectively. The NIRS-aided procedure proposed here can serve as an accurate, inexpensive, time-saving, and environment-friendly functional assay of tannin in Mediterranean browse. PMID- 14759162 TI - Effect of green tea supplementation on insulin sensitivity in Sprague-Dawley rats. AB - Epidemiological observations and laboratory studies have shown that green tea has a variety of health effects, including antitumor, antioxidative, and hypolipidemic activities. The aim of this study was to examine whether it had an effect on glucose tolerance and insulin sensitivity in Sprague-Dawley rats. In experiment 1 (in vivo study), rats were divided into two groups: a control group fed standard chow and deionized distilled water and a "green tea" group fed the same chow diet but with green tea instead of water (0.5 g of lyophilized green tea powder dissolved in 100 mL of deionized distilled water). After 12 weeks of green tea supplementation, the green tea group had lower fasting plasma levels of glucose, insulin, triglyceride, and free fatty acid than the control rats. Insulin-stimulated glucose uptake of, and insulin binding to, adipocytes were significantly increased in the green tea group. In experiment 2 (in vitro study), a tea polyphenol extract was used to determine its effect on insulin activity in vitro. Green tea polyphenols (0.075%) significantly increased basal and insulin stimulated glucose uptake of adipocytes. Results demonstrated that green tea increases insulin sensitivity in Sprague-Dawley rats and that green tea polyphenol is one of the active components. PMID- 14759163 TI - Excess electron-transfer-based repair of a Cis-Syn thymine dimer in DNA is not sequence dependent. AB - The repair of a cis-syn pyrimidine dimer by single electron reduction is possible in DNA over large distances. The intervening base sequence does not influence the repair yield. PMID- 14759164 TI - Mechanistic studies of the ethylene trimerization reaction with chromium diphosphine catalysts: experimental evidence for a mechanism involving metallacyclic intermediates. AB - A system for catalytic trimerization of ethylene utilizing CrCl3(THF)3 and a diphosphine ligand PNPOMe [= (o-MeO-C6H4)2PN(Me)P(o-MeO-C6H4)2] has been investigated. The coordination chemistry of chromium with PNPOMe has been explored, and (PNPOMe)CrCl3 and (PNPOMe)CrPh3 (3) have been synthesized by ether displacement from chromium(III) precursors. Salt metathesis of (PNPOMe)CrCl3 with o,o'-biphenyldiyl Grignard affords (PNPOMe)Cr(o,o'-biphenyldiyl)Br (4). Activation of 3 with H(Et2O)2B[C6H3(CF3)2]4 or 4 with NaB[C6H3(CF3)2]4 generates a catalytic system and trimerizes a 1:1 mixture of C2D4 and C2H4 to give isotopomers of 1-hexene without H/D scrambling (C6D12, C6D8H4, C6D4H8, and C6H12 in a 1:3:3:1 ratio). The lack of crossover supports a mechanism involving metallacyclic intermediates. The mechanism of the ethylene trimerization reaction has also been studied by the reaction of trans-, cis-, and gem-ethylene-d2 with 4 upon activation with NaB[C6H3(CF3)2]4. PMID- 14759165 TI - Effect of molecular oxygen on the variable-temperature 29Si MAS NMR spectra of zeolite-sorbate complexes. AB - Structure determinations of siliceous zeolite-sorbate host-guest complexes by solid-state NMR require highly resolved 29Si MAS NMR spectra. As the temperature is lowered, the 29Si MAS NMR spectra of many zeolite-sorbate complexes become broadened such that the resolution of the individual 29Si peaks is lost, limiting the application of solid-state NMR for structure determination. It is shown that the 29Si peak widths are related to the 29Si T2 relaxation times and that the source of the 29Si relaxation and the line broadening is paramagnetic molecular oxygen in the channels of the zeolite. Removal of the oxygen by purging the sample with nitrogen gas leads to a dramatic increase in the resolution of the 29Si MAS NMR spectrum of the p-dibromobenzene/ZSM-5 complex. An analysis of the individual 29Si T1 relaxation times reveals that the oxygen molecules are localized mainly in the zigzag channels of ZSM-5, suggesting that the p dibromobenzene molecules are located in the channel intersections. PMID- 14759166 TI - Microporous metal organic materials: promising candidates as sorbents for hydrogen storage. AB - Advancement in hydrogen storage techniques represents one of the most important areas of today's materials research. While extensive efforts have been made to the existing techniques, there is no viable storage technology capable of meeting the DOE cost and performance targets at the present time. New materials with significantly improved hydrogen adsorption capability are needed. Microporous metal coordination materials (MMOM) are promising candidates for use as sorbents in hydrogen adsorption. These materials possess physical characteristics similar to those of single-walled carbon nanotubes (SWNTs) but also exhibit a number of improved features. Here, we report a novel MMOM structure and its room temperature hydrogen adsorption properties. PMID- 14759167 TI - Synthesis of the epoxyquinol dimer RKB-3564 D: utilization of an alkoxysilanol to promote [4 + 4] dimerization. AB - The synthesis of the epoxyquinol dimer RKB-3564 D (3) is reported, employing an alkoxysilanol protecting group to redirect the inherently favored [4 + 2] dimerization of 2H-pyran monomers to a [4 + 4] manifold. Preliminary mechanistic studies indicate that the [4 + 4] dimerization may occur through a stepwise, ionic process. PMID- 14759168 TI - Tetranuclear rare earth metal polyhydrido complexes composed of "(C5Me4SiMe3)LnH2" Units. Unique reactivities toward unsaturated C-C, C-N, and C O bonds. AB - The tetranuclear Lu and Y polyhydrido complexes [(C5Me4SiMe3)Ln(mu-H)2]4(THF) (Ln = Lu, Y) undergo novel multiple hydrogenation reactions with unsaturated organic compounds such as benzonitrile, gamma-butyrolactone, styrene, and 1,4 bis(trimethylsilyl)-1,3-butadiyne to afford a series of structurally characterizable polynuclear complexes that possess novel structures and are otherwise difficult to access. Most of these reactions are unprecedented and can be attributed to the unique cooperative effects of multiple active sites in the polyhydrido rare earth metal complexes. PMID- 14759169 TI - Evidence for slow motion in proteins by multiple refocusing of heteronuclear nitrogen/proton multiple quantum coherences in NMR. AB - A novel NMR method characterizes slow motions in proteins by multiple refocusing of double- and zero-quantum coherences of amide protons and nitrogen-15 nuclei. If both nuclei experience changes in their isotropic chemical shifts because of internal motions on slow time scales (mus - ms), this leads to a difference in the relaxation rates of double- and zero-quantum coherences. This is due to CSM/CSM (chemical shift modulation) cross-correlation effects that are related to the well-known chemical exchange contribution Rex to the decay rate R2 = 1/T2 of nitrogen-15 nuclei. The CSM/CSM contributions can be distinguished from other mechanisms through their dependence on the repetition rate of a Carr-Purcell Meiboom-Gill (CPMG) multiple refocusing sequence. In ubiquitin, motional processes can be identified that could hitherto not be observed by conventional CPMG nitrogen-15 NMR. PMID- 14759170 TI - Direct chemical evidence for charge transfer between photoexcited 2-aminopurine and guanine in duplex DNA. AB - Photoexcited 2-aminopurine (Ap*) is extensively exploited as a fluorescent base analogue in the study of DNA structure and dynamics. Quenching of Ap* in DNA is often attributed to stacking interactions between Ap* and DNA bases, despite compelling evidence indicating that charge transfer (CT) between Ap* and DNA bases contributes to quenching. Here we present direct chemical evidence that Ap* undergoes CT with guanine residues in duplex DNA, generating oxidative damage at a distance. Irradiation of Ap in DNA containing the modified guanine, cyclopropylguanosine (CPG), initiates hole transfer from Ap* followed by rapid ring opening of the CPG radical cation. Ring opening accelerates hole trapping to a much shorter time regime than for guanine radicals in DNA; consequently, trapping effectively competes with back electron transfer (BET) leading to permanent CT chemistry. Significantly, BET remains competitive, even with this much faster trapping reaction, consistent with measured kinetics of DNA-mediated CT. The distance dependence of BET is sharper than that of forward CT, leading to an inverted dependence of product yield on distance; at short distances product yield is inhibited by BET, while at longer distances trapping dominates, leading to permanent products. The distance dependence of product yield is distinct from forward CT, or charge injection. As with photoinduced charge transfer in other chemical and biological systems, rapid kinetics for charge injection into DNA need not be associated with a high yield of DNA damage products. PMID- 14759171 TI - Influence of architecture on the kinetic stability of molecular assemblies. AB - The strength of a multimolecular system depends on the number of interactions that hold it together. Using dynamic force spectroscopy, we show how the kinetic stability of a system decreases as the number of molecular bonds is increased, as predicted by theory. The data raise important considerations for experimental tests of bond strength and, as a paradigm, suggest both routes to and pitfalls in methods for computational simulation of molecular transitions, such as ligand binding and protein folding. PMID- 14759172 TI - A zeolitic organo-metallophosphate hybrid material with bimodal porosity. AB - The first metal carboxylatophosphate, NTHU-2, contains inorganic ZnHPO4 layers linked by BDC units (BDC = 1,4-benzene dicarboxylate or terephthalic anion); the three-dimensional anionic framework has large pores with the smallest diameter being 1.36 nm; N2 sorption isotherms reveal both micro- and mesoporosity; the new material is photoluminescent and disassembles in water wherein the discharged organic fragments form mixed crystals. PMID- 14759173 TI - Self-assembled spiral nanoarchitecture and supramolecular chirality in Langmuir Blodgett films of an achiral amphiphilic barbituric acid. AB - An amphiphilic barbituric acid derivative was found to form stable monolayers showing a clear phase transition at the air/water interface. It is interesting to find that the deposited Langmuir-Blodgett (LB) films of the compound showed circular dichroism (CD) although the molecule itself was achiral. AFM measurements on the transferred one-layer LB film revealed that spiral nanoarchitectures were formed. It was further found that the supramolecular chirality of the LB films was related to symmetry breaking at the interface. Hydrogen bonding and the pi-pi stacking between the neighboring molecules resulted in chiral fibers which formed the spiral structures. To the best of our knowledge, this is the first report on the chirality of the molecular assemblies and spiral nanostructures formed through the air/water interface by achiral molecules. PMID- 14759174 TI - Near-complete suppression of quantum dot blinking in ambient conditions. AB - Colloidal semiconductor quantum dots are attractive fluorophores for multicolor imaging because of broad absorption and narrow emission spectra, and they are brighter and far more photostable than organic dyes. However, severe intermittence in emission (also known as blinking) has been universally observed from single dots and has been considered an intrinsic limitation difficult to overcome. This is unfortunate because growing applications in spectroscopy of single biological molecules and quantum information processing using single photon sources could greatly benefit from long-lasting and nonblinking single molecule emitters. For instance, in a recent application of single-dot imaging, the tracking of membrane receptors was interrupted frequently due to the stroboscopic nature of recording. Blinking can also reduce the brightness in ensemble imaging via signal saturation. Here we show that the quantum dot blinking can be suppressed with the emission duty cycle approaching 100% while maintaining biocompatibility. PMID- 14759175 TI - Isolation and ring-opening of new 1-sila-3-metallacyclobutanes (eta 5 C5H4Fe)(CO)2CH2SiR2 leading to a new class of organometallic polymer. AB - Treatment of FpCH2SiR2Cl [Fp = (eta5-C5H5)Fe(CO)2] with lithium diisopropylamide leads to the formation of new metallacycle complexes. When R2 = Me2 and MenBu, these ring-open to form a new class of organometallic polymer. When R2 = nBu2, the metallacycle resists ring-opening and may be structurally characterized as the Ph3P-substituted species. PMID- 14759176 TI - In Rhodobacter sphaeroides respiratory nitrate reductase, the kinetics of substrate binding favors intramolecular electron transfer. AB - The respiratory nitrate reductase (NapAB) from Rb. sphaeroides is a periplasmic molybdenum-containing enzyme which belongs to the DMSO reductase family. We report a study of NapAB by protein film voltammetry (PFV), and we present the first quantitative interpretation of the complex redox-state dependence of activity that has also been observed with other related enzymes. The model we use to fit the data assumes that binding of substrate partly limits turnover and is faster and weaker when the Mo ion is in the V oxidation state than when it is fully reduced. We explain how the presence in the catalytic cycle of such slow chemical steps coupled to electron transfer to the active site decreases the driving force required to reduce the MoV ion and makes exergonic the last intramolecular electron-transfer step (between the proximal cubane and the Mo cofactor). Importantly, comparison is made with all Mo enzymes for which PFV data are available, and we emphasize general features of the energetics of the catalytic cycles in enzymes of the DMSO reductase family. PMID- 14759177 TI - An unprecedented hybrid scorpionate/cyclopentadienyl ligand. AB - An efficient method for the preparation of the first hybrid scorpionate/cyclopentadienyllithium compound as a new class of tridentate ligand is described. This compound is an excellent reagent for the introduction of this ligand into transition metal complexes. PMID- 14759178 TI - Fabrication of metal structures with nanometer-scale lateral dimensions by sectioning using a microtome. AB - This Communication describes the fabrication of planar structures comprising metallic features with nanometer-scale lateral dimensions in polymer prepared by sectioning a thin metallic film, embedded in a polymer matrix, in a plane perpendicular to the metallic film. This procedure converts a structure that is thin along the z-axis into a structure that is thin along the x-axis. The embedded thin metal film is still conductive and can be used as a nanoelectrode. The structure and composition of the exposed nanoedge can be easily tuned by changing the structure of the surface supporting the metal film, and the composition and the thickness of that film. PMID- 14759179 TI - Catalytic dehydrocoupling of amine-borane and phosphine-borane adducts: the mechanism is heterogeneous in one case and homogeneous in the other. AB - The catalytic dehydrocoupling reactions of Me2NH.BH3 and Ph2PH.BH3 using the rhodium precatalyst [Rh(1,5-cod)(mu-Cl)]2 were found to proceed by different mechanisms: heterogeneous involving Rh(0) metal for the former case and homogeneous for the latter. PMID- 14759180 TI - Stereoselective palladium-catalyzed O-glycosylation using glycals. AB - A highly stereoselective palladium-catalyzed O-glycosylation reaction is described. The reaction of a glycal 3-acetate or carbonate with the zinc(II) alkoxide of acceptors establishes the glycosidic linkage under palladium catalysis to give rise to disaccharides as the product in good yields and with high stereoselectivity. In contrast to the Lewis acid mediated Ferrier procedure, the anomeric stereochemistry of this reaction is controlled by the employed ligand. Whereas the use of a complex of palladium acetate and 2-di(tert butyl)phosphinobiphenyl as the catalyst results in the exclusive beta-glycoside formation, the same reaction using trimethyl phosphite ligand furnishes an alpha anomer as the major product. The utility of the 2,3-unsaturation present in the resulting glycoside is demonstrated by the further transformations such as dihydroxylation, hydration, and hydrogenation reactions. Thus, the combination of the glycosylation and subsequent functionalization provides a novel entry to saccharides which are otherwise difficult to prepare. The broad scope of the process, mildness of the reaction conditions, and experimental simplicity should make this method a useful tool in synthetic carbohydrate chemistry. PMID- 14759181 TI - Tellurium-mediated cycloaromatization of acyclic enediynes under mild conditions. AB - The cycloaromatization of acyclic enediynes typically requires very high temperatures (>160 degrees C) and dilute conditions to proceed in a synthetically useful yield. These conditions hinder reaction throughput, inhibiting the use of this reaction for the large-scale production of materials. The reaction of sodium telluride with acyclic arenediynes yields the corresponding tellurepine, which under gentle heating extrudes Te degrees to yield the cycloaromatization product. We have developed conditions that form sodium telluride from inexpensive tellurium metal in situ, and that also perform the desilylation of silylated arenediynes in the same process. Under our conditions, we are able to perform desilylation and cycloaromatization at temperatures as low as 40 degrees C and on a scale as large as 5 g in standard laboratory glassware. PMID- 14759182 TI - Suzuki cross-couplings of unactivated secondary alkyl bromides and iodides. AB - The capacity to employ unactivated alkyl electrophiles as coupling partners will markedly increase the already exceptional utility of metal-catalyzed cross coupling processes. This communication describes the development of a method that achieves the first Suzuki reactions of unactivated secondary alkyl bromides and iodides. The ability to couple readily available, easy-to-handle boronic acids is an attractive feature of this catalyst system. PMID- 14759183 TI - Amino-aryl-carbenes: alternative ligands for transition metals? AB - Despite the presence of a single amino substituent, an amino-anthryl-carbene was found to behave as a strong sigma-donor weak pi-acceptor ligand toward rhodium(I) fragments. PMID- 14759184 TI - Radical-type reactivity of the 1,3-dibora-2,4-diphosphoniocyclobutane-1,3-diyl. AB - Despite through-space and through-bond B-B interactions, the stable 1,3-bora-2,4 diphosphoniocyclobutane-1,3-diyl displays some radical-type behavior, as illustrated by the spontaneous formation of the trans 1,3-adduct with trimethyltin hydride and a B-spiro derivative with bromotrichloromethane. PMID- 14759185 TI - A sulfoxide-promoted, catalytic method for the regioselective synthesis of allylic acetates from monosubstituted olefins via C-H oxidation. AB - Sulfoxide ligation to Pd(II) salts is shown to selectively promote C-H oxidation versus Wacker oxidation chemistry and to control the regioselectivity in the C-H oxidation products. A catalytic method for the direct C-H oxidation of monosubstituted olefins to linear (E)-allylic acetates in high regio- and stereoselectivities and preparatively useful yields is described. The method using benzoquinone as the stoichiometric oxidant and 10 mol % of Pd(OAc)2 or Pd(O2CCF3)2 as the catalyst in a DMSO/AcOH (1:1) solution was found to be compatible with a wide range of functionality (e.g., amides, carbamates, esters, and ethers, see Table 2). Addition of DMSO was found to be critical for promoting the C-H oxidation pathway, with AcOH alone or in combination with a diverse range of dielectric media, leading to mixtures favoring Wacker-type oxidation products (Tables 1, S3). To explore the role of DMSO as a ligand, the bis-sulfoxide Pd(OAc)2 complex 1 was formed and found to be an effective C-H oxidation catalyst in the absence of DMSO (eqs 2, 3). Moreover, catalyst 1 effects a reversal of regioselectivity, favoring the formation of branched allylic acetates. PMID- 14759186 TI - Controlled photoinitiated release of nitric oxide from polymer films containing S nitroso-N-acetyl-DL-penicillamine derivatized fumed silica filler. AB - This report describes the first hydrophobic nitric oxide (NO)-releasing material that utilizes light as an external on/off trigger to control the flux of NO generated from cured polymer films. Fumed silica polymer filler particles were derivatized with S-nitroso-N-acetyl-dl-penicillamine and blended into the center layer of trilayer silicone rubber films. Nitric oxide is generated upon irradiation with light, and fluxes increase with increasing power of incident light. The ability to precisely control NO generation from this material has the potential to answer fundamental questions about the levels of NO needed to achieve desired therapeutic affects in different biomedical applications. PMID- 14759187 TI - Nucleophilic attack by oxyanions on a phosphate monoester dianion: the positive effect of a cationic general Acid. AB - The two negative charges on a phosphate monoester RO-PO32- at neutral pH provide a considerable electrostatic barrier toward reactions with nucleophilic reagents with a negative charge on the attacking atom. Electrostatic repulsion disappears when the hydrolysis of an aryl phosphate monoester is catalyzed by a neighboring cationic general acid. The hydrolysis of 8-dimethylammonium-1-phosphate (1) is catalyzed by oxyanions, fluoride anion, and hydroxylamines at similar rates. PMID- 14759188 TI - Rapid assembly of polycyclic substances by a multicomponent cascade (4 + 2)-(2 + 2) cycloadditions: total synthesis of the proposed structure of paesslerin A. AB - A versatile method for stereoselective formation of multisubstituted bicyclo[4.2.0]octane framework has been developed by means of catalytic (4 + 2) (2 + 2) cycloaddition reactions. Its application to the synthesis of a substance reported to be the cytotoxic sesquiterpene, paesslerin A, is described, and it has been made clear that a revision of the structure of natural paesslerin A is required. PMID- 14759189 TI - Triplet pi-diradical dication of directly meso-beta-linked porphyrin dimer. AB - Directly linked bisporphyrin, Zn(II)2 5,10,15,20-tetrakis(2,4,6-trimethylphenyl) 2-(10,15,20-tris(3,5-di-tert-butylphenyl)porphyrin-5-yl)porphyrin, was investigated on the basis of the magnetic property of its bis(pi-radical cation). A stable triplet state of the bis(pi-radical cation) of the bisporphyrin, which obeys the Curie law, was successfully observed. PMID- 14759190 TI - Molecular dynamics simulations of gas diffusion in metal-organic frameworks: argon in CuBTC. AB - The class of coordination polymers known as metal-organic frameworks (MOFs) has three-dimensional porous structures that are considered as a promising alternative to zeolites and other nanoporous materials for catalysis, gas adsorption, and gas separation applications. In this paper, we present the first study of gas diffusion inside an MOF and compare the observed diffusion to known behaviors in zeolites. Using grand canonical Monte Carlo and equilibrium molecular dynamics, we calculate the adsorption isotherm and self-, corrected, and transport diffusivities for argon in the CuBTC metal-organic framework. Our results indicate that diffusion of Ar in CuBTC is very similar to Ar diffusion in silica zeolites in magnitude, concentration, and temperature dependence. This conclusion appears to apply to a broad range of MOF structures. PMID- 14759191 TI - Stereocontrolled total synthesis of (-)-vincamajinine and (-)-11-methoxy-17 epivincamajine. AB - The first total syntheses of (-)-vincamajinine (5) (from Na-methyl-d-tryptophan methyl ester) and (-)-11-methoxy-17-epivincamajine (6) (from Na-methyl-6-methoxy d-tryptophan ethyl ester) are described. The syntheses have been completed in a highly stereocontrolled manner (>98% ee). Key steps included the asymmetric Pictet-Spengler reaction, enolate-mediated palladium cross-coupling reaction, and acid-catalyzed formation of the C(7)-C(17) bond. In addition, the triethylsilane/TFA-mediated incorporation of the 2alpha-H (11 to 12) and the borohydride generation of the C(17) hydroxyl function (R) were also stereospecific. The unique highly conjested carbon skeletons of the two alkaloids were completed in a concise manner and in regiospecific fashion. PMID- 14759192 TI - Mechanistic investigation leads to a synthetic improvement in the hydrolytic kinetic resolution of terminal epoxides. AB - The mechanism of the hydrolytic kinetic resolution (HKR) of terminal epoxides was investigated by kinetic analysis using reaction calorimetry. The chiral (salen)Co X complex (X = OAc, OTs, Cl) undergoes irreversible conversion to (salen)Co-OH during the course of the HKR and thus serves as both precatalyst and cocatalyst in a cooperative bimetallic catalytic mechanism. This insight led to the identification of more active catalysts for the HKR of synthetically useful terminal epoxides. PMID- 14759193 TI - Mapping enzyme active sites in complex proteomes. AB - Genome sequencing projects have uncovered many novel enzymes and enzyme classes for which knowledge of active site structure and mechanism is limited. To facilitate mechanistic investigations of the numerous enzymes encoded by prokaryotic and eukaryotic genomes, new methods are needed to analyze enzyme function in samples of high biocomplexity. Here, we describe a general strategy for profiling enzyme active sites in whole proteomes that utilizes activity-based chemical probes coupled with a gel-free analysis platform. We apply this gel-free strategy to identify the sites of labeling on enzymes targeted by sulfonate ester probes. For each enzyme examined, probe labeling was found to occur on a conserved active site residue, including catalytic nucleophiles (e.g., C32 in glutathione S-transferase omega) and bases/acids (e.g., E269 in aldehyde dehydrogenase-1; D204 in enoyl CoA hydratase-1), as well as residues of unknown function (e.g., D127 in 3 beta-hydroxysteroid dehydrogenase/isomerase-1). These results reveal that sulfonate ester probes are remarkably versatile activity based profiling reagents capable of labeling a diversity of catalytic residues in a range of mechanistically distinct enzymes. More generally, the gel-free strategy described herein, by consolidating into a single step the identification of both protein targets of activity-based probes and the specific residues labeled by these reagents, provides a novel platform in which the proteomic comparison of enzymes can be accomplished in unison with a mechanistic analysis of their active sites. PMID- 14759194 TI - Dynamics of an enzymatic substitution reaction in haloalkane dehalogenase. AB - Reactive flux molecular dynamics simulations have been carried out using a combined QM/MM potential to study the dynamics of the nucleophilic substitution reaction of dichloroethane by a carboxylate group in haloalkane dehalogenase and in water. We found that protein dynamics accelerates the reaction rate by a factor of 2 over the uncatalyzed reaction. Compared to the thermodynamic effect in barrier reduction, protein dynamic contribution is relatively small. However, analyses of the friction kernel reveal that the origins of the reaction dynamics in water and in the enzyme are different. In aqueous solution, there is significant electrostatic solvation effect, which is reflected by the slow reorganization relaxation of the solvent. On the other hand, there is no strong electrostatic coupling in the enzyme and the major effect on reaction coordinate motion is intramolecular energy relaxation. PMID- 14759195 TI - Total chemical synthesis of crambin. AB - Crambin is a small (46 amino acids) protein isolated from the seeds of the plant Crambe abyssinica. Crambin has been extensively used as a model protein for the development of advanced crystallography and NMR techniques and for computational folding studies. We set out to establish synthetic access to crambin. Initially, we synthesized the 46 amino acid polypeptide by native chemical ligation of two distinct sets of peptide segments (15 + 31 and 31 + 15 residues). The synthetic polypeptide chain folded in good yield to give native crambin containing three disulfide bonds. The chemically synthesized crambin was characterized by LC-MS and by 2D-NMR. However, the 31-residue peptide segments were difficult to purify, and this caused an overall low yield for the synthesis. To overcome this problem, we synthesized crambin by the native chemical ligation of three segments (15 + 16 + 15 residues). Total synthesis using the ligation of three segments gave more than a 10-fold increase in yield and a protein product of exceptionally high purity. This work demonstrates the efficacy of chemical protein synthesis by the native chemical ligation of three segments and establishes efficient synthetic access to the important model protein crambin for experimental studies of protein folding and stability. PMID- 14759196 TI - Optical sensors based on hybrid aptamer/conjugated polymer complexes. AB - Single-stranded DNA (aptamer) can specifically bind potassium ions or human alpha thrombin. When binding takes place, the aptamer undergoes a conformational transition from an unfolded to a folded structure. This conformational change of the negatively charged oligonucleotide can be detected by adding a water-soluble, cationic polythiophene derivative, which transduces the new complex formation into an optical (colorimetric or fluorometric) signal without any labeling of the probe or of the target. This simple and rapid methodology has enabled the detection of human thrombin in the femtomole range. This new biophotonic tool can easily be applied to the detection of various other proteins as well as being useful in the high-throughput screening of new drugs. PMID- 14759197 TI - Donor-acceptor (electronic) coupling in the precursor complex to organic electron transfer: intermolecular and intramolecular self-exchange between phenothiazine redox centers. AB - Intermolecular electron transfer (ET) between the free phenothiazine donor (PH) and its cation radical (PH*+) proceeds via the [1:1] precursor complex (PH)(2)*+ which is transiently observed for the first time by its diagnostic (charge resonance) absorption band in the near-IR region. Similar intervalence (optical) transitions are also observed in mixed-valence cation radicals with the generic representation: P(br)P*+, in which two phenothiazine redox centers are interlinked by p-phenylene, o-xylylene, and o-phenylene (br) bridges. Mulliken Hush analysis of the intervalence (charge-resonance) bands afford reliable values of the electronic coupling element H(IV) based on the separation parameters for (P/P*+) centers estimated from some X-ray structures of the intermolecular (PH)(2)*+ and the intramolecular P(br)P*+ systems. The values of H(IV), together with the reorganization energies lambda derived from the intervalence transitions, yield activation barriers DeltaG(ET)() and first-order rate constants k(ET) for electron-transfer based on the Marcus-Hush (two-state) formalism. Such theoretically based values of the intrinsic barrier and ET rate constants agree with the experimental activation barrier (E(a)) and the self exchange rate constant (k(SE)) independently determined by ESR line broadening measurements. This convergence validates the use of the two-state model to adequately evaluate the critical electronic coupling elements between (P/P*+) redox centers in both (a) intermolecular ET via the precursor complex and (b) intramolecular ET within bridged mixed-valence cation radicals. Important to intermolecular ET mechanism is the intervention of the strongly coupled precursor complex since it leads to electron-transfer rates of self-exchange that are 2 orders of magnitude faster (and activation barrier that is substantially lower) than otherwise predicted solely on the basis of Marcus reorganization energy. PMID- 14759198 TI - A hexa-peri-hexabenzocoronene cyclophane: an addition to the toolbox for molecular electronics. AB - Cyclophanes with the largest-to-date polycyclic aromatic hydrocarbon (hexa-peri hexabenzocoronene, HBC) to be entrained in such a structural motif are reported. The two disks are covalently captured by intermolecular ring-closing olefin metathesis of dienes in good yield. DSC, optical microscopy, and WAXD show the new cyclophanes to self-assemble to thermotropic columnar liquid crystal mesophases similar to monomeric analogues. Solution spectroscopic studies reveal that the two disks within a single unit lie face-to-face, with a small average lateral offset. Self-assembly into two-dimensional crystals at a solid-liquid interface was visualized by STM, and the electrical properties of single molecules were assessed by scanning tunneling spectroscopy revealing a diode-like behavior which is similar to that previously reported for single HBC disks, laying the groundwork for future electrical interrogations of dynamic molecular complexes. PMID- 14759199 TI - CD spectra of polynuclear complexes of diimine ligands: theoretical and experimental evidence for the importance of internuclear exciton coupling. AB - We have recently reported on dinuclear complexes Lambda,Lambda [Co(2)L(2)Cl(2)]CoCl(4) of two novel chiral ligands (1a and 1b) which contain pyridyl-imine chelate groups (Telfer, S. G.; Sato, T.; Kuroda, R. Chem. Commun. 2003, 1064-1065). The absolute configuration of the cobalt(II) centers was unambiguously assigned by X-ray crystallography. However, the sign of the exciton couplets in their CD spectra was opposite to that expected on the basis of the stereochemistry of the metal centers. We present a rationalization of these anomalous spectra in terms of an "internuclear" exciton coupling model which takes into account the coupling of chromophores located on different metal centers. We have performed a series of semiempirical (ZINDO) calculations which provide quantitative support to this model. These findings show that the absolute configuration of the metal centers in a polynuclear complex may be incorrectly assigned on the basis of CD data if internuclear coupling effects are not taken into consideration. We summarize the CD spectral data of number of other chiral polynuclear complexes from the literature, including dinuclear complexes bridged by the 2,2'-bipyrimidine ligand, complexes of the HAT ligand, and dinuclear triple-stranded helicates. The amplitude of the CD spectra of many of these complexes is not additive with the number of chromophores. These anomalous spectra can be accounted for by taking internuclear coupling effects into consideration. PMID- 14759200 TI - From achiral ligands to chiral coordination polymers: spontaneous resolution, weak ferromagnetism, and topological ferrimagnetism. AB - Using the achiral diazine ligands bearing two bidentate pyridylimino groups as sources of conformational chirality, five azido-bridged coordination polymers are prepared and characterized crystallographically and magnetically. The chirality of the molecular units is induced by the coordination of the diazine ligands in a twisted chiral conformation. The use of L(1) (1,4-bis(2-pyridyl)-1-amino-2,3 diaza-1,3-butadiene) and L(2) (1,4-bis(2-pyridyl)-1,4-diamino-2,3-diaza-1,3 butadiene) induces spontaneous resolution, yielding conglomerates of chiral compounds [Mn(3)(L(1))(2)(N(3))(6)](n) (1) and [Mn(2)(L(2))(2)(N(3))(3)](n)(ClO(4))(n).nH(2)O (2), respectively, where triangular (1) or double helical (2) chiral units are connected into homochiral one-dimensional (1D) chains via single end-to-end (EE) azido bridges. The chains are stacked via hydrogen bonds in a homochiral fashion to yield chiral crystals. When L(3) (2,5-bis(2-pyridyl)-3,4-diaza-2,4-hexadiene) is employed, a partial spontaneous resolution occurs, where binuclear chiral units are interlinked into fish-scale-like homochiral two-dimensional (2D) layers via single EE azido bridges. The layers are stacked in a heterochiral or homochiral fashion to yield simultaneously a racemic compound, [Mn(2)(L(3))(N(3))(4)](n) (3a), and a conglomerate, [Mn(2)(L(3))(N(3))(4)](n).nMeOH (3b). On the other hand, the ligand without amino and methyl substituents (L(4), 1,4-bis(2-pyridyl)-2,3-diaza-1,3 butadiene) does not induce spontaneous resolution. The resulting compound, [Mn(2)(L(4))(N(3))(4)](n) (4), consists of centrosymmetric 2D layers with alternating single diazine, single EE azido, and double end-on (EO) azido bridges, where the chirality is destroyed by the centrosymmetric double EO bridges. These compounds exhibit very different magnetic behaviors. In particular, 1 behaves as a metamagnet built of homometallic ferrimagnetic chains with a unique "fused-triangles" topology, 2 behaves as a 1D antiferromagnet with alternating antiferromagnetic interactions, 3a and 3b behave as spin-canted weak ferromagnets with different critical temperatures, and 4 also behaves as a spin canted weak ferromagnet but exhibits two-step magnetic transitions. PMID- 14759201 TI - Selective activation of two sites in RNA by acridine-bearing oligonucleotides for clipping of designated RNA fragments. AB - Artificial enzymes for selective scission of RNA at two designated sites, which are valuable for advanced RNA science, have been prepared by combining lanthanide(III) ion with an oligonucleotide bearing two acridine groups. When these modified oligonucleotides form heteroduplexes with substrate RNA, the two phosphodiester linkages in front of the acridines are selectively activated and preferentially hydrolyzed by lanthanide ion. This two-site RNA scission does not require any specific RNA sequence at the scission sites, and the length of clipped RNA fragment is easily and precisely controllable by changing the distance between two acridine groups in the modified oligonucleotide. The two site scission is also successful even when the substrate RNA has higher-order structures. By using these two-site RNA cutters, RNA fragments of predetermined length were obtained from long RNA substrates and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). Single nucleotide polymorphisms in homozygous and heterozygous samples were accurately and easily detected in terms of the difference in mass number. Multiplex analyses of in vitro transcripts from human genome were also successful. PMID- 14759202 TI - Scope of palladium-catalyzed alkylative ring opening. AB - We have explored the scope of the palladium-catalyzed nucleophilic ring opening methodology. New highly selective and highly active catalysts have been found for the ring opening of oxabenzonorbornadienes. Employing these catalysts, the addition of various alkyl nucleophiles to oxabenzonorbornadiene has been achieved. In addition, reaction of diethylzinc with [3.2.1] oxabicyclic alkenes has been accomplished to yield ring-opened products as well as functionalized alkene addition products. PMID- 14759203 TI - NOE and PGSE NMR spectroscopic studies of solution structure and aggregation in metallocenium ion-pairs. AB - The solution structures of the metallocenium homogeneous polymerization catalyst ion-pairs [Cp(2)ZrMe](+)[MeB(C(6)F(5))(3)](-) (1), [(1,2 Me(2)Cp)(2)ZrMe](+)[MeB(C(6)F(5))(3)](-) (2), [(Me(2)SiCp(2))ZrMe](+)[MeB(C(6)F(5))(3)](-) (3), [Me(2)C(Fluorenyl)(Cp)ZrMe](+)[FPBA](-) (FPBA = tris(2,2',2' ' nonafluorobiphenyl)fluoroaluminate) (4), [rac-Et(Indenyl)(2)ZrMe](+)[FPBA](-) (5), [(Me(5)Cp)(2)ThMe](+)[B(C(6)F(5))(4)](-) (6), [(Me(2)SiCp(2))Zr(Me)(THF)](+)[MeB(C(6)F(5))(3)](-) (7), [(Me(2)SiCp(2))Zr(Me)(PPh(3))](+)[MeB(C(6)F(5))(3)](-) (8), [(Me(2)SiCp(2))Zr(Me)(THF)](+)[B(C(6)F(5))(4)](-) (9), [(Me(2)Si(Me(4)Cp)(t BuN)Zr(Me)(solvent)](+)[B(C(6)F(5))(4)](-) (solvent = benzene, toluene) (10), [(Cp(2)ZrMe)(2)(mu-Me)](+)[MePBB](-) (PBB = tris(2,2',2" nonafluorobiphenyl)borane) (11), and [(Cp(2)Zr)(2)(mu-CH(2))(mu-Me)](+)[MePBB](-) (12), having the counteranion in the inner (1, 3, 4, 5, and 6) or outer (7, 8, 9, 10, 11, and 12) coordination sphere, have been investigated for the first time in solvents with low relative permittivity such as benzene or toluene by (1)H NOESY and (1)H,(19)F HOESY NMR spectroscopy. It is found that the average interionic solution structures of the inner sphere contact ion-pairs are similar to those in the solid state with the anion B-Me (1, 3) or Al-F (5) vectors oriented toward the free zirconium coordination site. The HOESY spectrum of complex 6 is in agreement with the reported solid-state structure. In contrast, in outer sphere contact ion-pairs 7, 8, 9, and 10, the anion is located far from the Zr-Me(+) moiety and much nearer to the Me(2)Si bridge than in 3. The interionic structure of 8 is concentration-dependent, and for concentrations greater than 2 mM, a loss of structural localization is observed. PGSE NMR measurements as a function of concentration (0.1-5.0 mM) indicate that the tendency to form aggregates of nuclearity higher than simple ion-pairs is dependent on whether the anion is in the inner or outer coordination sphere of the metallocenium cation. Complexes 2, 3, 4, 5, and 6 show no evidence of aggregation up to 5 mM (well above concentrations typically used in catalysis) or at the limit of saturated solutions (complexes 3 and 6), while concentration-dependent behavior is observed for complexes 7, 8, 10, and 11. These outer sphere ion-pairs begin to exhibit significant evidence for ion-quadruples in solutions having concentrations greater than 0.5 mM with the tendency to aggregate being a function of metal ligation and anion structure. Above 2 mM, compound 8 exists as higher aggregates that are probably responsible for the loss of interionic structural specificity. PMID- 14759204 TI - Choice of coordination number in d10 complexes of group 11 metals. AB - The distribution of di-, tri-, and tetracoordination among the d(10) ions of the group 11 metals is theoretically analyzed by means of density functional calculations on more than 150 model complexes of general formula [MX(m)L(n)](1-m) (where M = Cu, Ag, or Au; L = NH(3) or PH(3); X = Cl, Br, or I; m + n = 2-4). The energy of a ligand association reaction has been found to be practically determined by two contributions: the interaction energy and the energy of deformation of the metal coordination sphere. The larger deformation energy of gold complexes compared to copper and silver ones explains the predominance of dicoordination among Au(I) complexes, in comparison with Cu(I) and Ag(I), for which dicoordination is far less common than tri- and tetracoordination. Other experimental trends can be explained by looking at the fine details of these two energetic components. PMID- 14759205 TI - Synthesis, structure and physical properties of the first one-dimensional phenalenyl-based neutral radical molecular conductor. AB - We report the preparation, crystallization, and solid-state characterization of a benzyl-substituted spirobiphenalenyl radical. The crystal structure shows that the radical is monomeric in the solid state, with the molecules packed in an unusual one-dimensional (1-D) fashion that we refer to as a pi-step stack. This particular mode of 1-D stacking is forced on the lattice arrangement by the presence of the orthogonal phenalenyl units that were specifically incorporated to prevent the crystallization of low-dimensional structures. The structure shows that this strategy is effective, and neighboring molecules in the stack can only interact via the overlap of one pair of active (spin-bearing) carbon atoms per phenalenyl unit, leading to the pi-step structure in which the remaining four active carbon atoms per phenalenyl unit do not interact with nearest neighbor molecules. The magnetic susceptibility data in the temperature range 4-360 K may be fit to an antiferromagnetic Heisenberg S = 1/2 linear chain model with intrachain spin coupling J = -52.3 cm(-1). Despite the uniform stacking, the material has a room temperature conductivity of 1.4 x 10(-3) S/cm and is best described as a Mott insulator. PMID- 14759206 TI - Electron transfer at self-assembled monolayers measured by scanning electrochemical microscopy. AB - New approaches have been developed for measuring the rates of electron transfer (ET) across self-assembled molecular monolayers by scanning electrochemical microscopy (SECM). The developed models can be used to independently measure the rates of ET mediated by monolayer-attached redox moieties and direct ET through the film as well as the rate of a bimolecular ET reaction between the attached and dissolved redox species. By using a high concentration of redox mediator in solution, very fast heterogeneous (10(8) s(-1)) and bimolecular (10(11) mol(-1) cm(3) s(-1)) ET rate constants can be measured. The ET rate constants measured for ferrocene/alkanethiol on gold were in agreement with previously published data. The rates of bimolecular heterogeneous electron transfer between the monolayer-bound ferrocene and water-soluble redox species were measured. SECM was also used to measure the rate of ET through nonelectroactive alkanethiol molecules between substrate gold electrodes and a redox probe (Ru(NH(3))(6)(3+)) freely diffusing in the solution, yielding a tunneling decay constant, beta, of 1.0 per methylene group. PMID- 14759207 TI - Nanogel nanosecond photonic crystal optical switching. AB - We developed a robust nanosecond photonic crystal switching material by using poly(N-isopropylacrylamide) (PNIPAM) nanogel colloidal particles that self assemble into crystalline colloidal arrays (CCAs). The CCA was polymerized into a loose-knit hydrogel which permits the individual embedded nanogel PNIPAM particles to coherently and synchronously undergo their thermally induced volume phase transitions. A laser T-jump from 30 to 35 degrees C actuates the nanogel particle shrinkage; the resulting increased diffraction decreases light transmission within 900 ns. Additional transmission decreases occur with characteristic times of 19 and 130 ns. Individual NIPAM sphere volume switching occurs in the approximately 100 ns time regime. These nanogel nanosecond phenomena may be useful in the design of fast photonic crystal switches and optical limiting materials. Smaller nanogels will show even faster volume phase transitions. PMID- 14759208 TI - New approach to oligonucleotide microarrays using zirconium phosphonate-modified surfaces. AB - A new approach to oligonucleotide arrays is demonstrated that utilizes zirconium phosphonate-derivatized glass slides. The active slides are prepared by binding Zr(4+) to surfaces terminated with organophosphonate groups previously deposited using either Langmuir-Blodgett or self-assembled monolayer methods. Oligonucleotide probes modified with a terminal phosphate bind strongly to the active zirconium phosphonate monolayer, and arrays for detecting fluorescent targets have been prepared using commercial spotting and scanning instruments. Preferred binding to the surface of the terminal phosphate of the modified probes instead of the internal phosphate diester groups is demonstrated and shown to yield increased fluorescence intensity after hybridization with labeled targets. A significant decrease in background signal is achieved by treating the slides with bovine serum albumin after spotting and before hybridization. A further increase in fluorescence after hybridization is observed when using a poly guanine spacer between the probe oligomer and the terminal phosphate. Combining these modifications, an intensity ratio of nearly 1000 is achieved when comparing 5'-phosphate-modified 33-mer probes with unmodified probes upon hybridization with fluorescent targets. PMID- 14759209 TI - Silver-phosphine complexes of the highly methylated carborane monoanion [closo-1 H-CB11Me11]-. AB - The synthesis of the silver(I) salt of the highly methylated carborane anion [closo-1-H-CB(11)Me(11)](-) is described, Ag[closo-1-H-CB(11)Me(11)] 1, which in the solid state shows close intermolecular Ag...H(3)C contacts. Addition of various monodentate phosphines to 1 results in the formation of the complexes (R(3)P)Ag[closo-1-H-CB(11)Me(11)] [R = Ph, 2; cyclohexyl (C(6)H(11)), 3; (3,5 Me(2)-C(6)H(3)), 4]. All these complexes show close intermolecular Ag.H(3)C contacts in the solid state that are considerably shorter than the sum of the van der Waals radius of methyl (2.00 A) and the ionic radius of silver(I) (1.29 A). For 2 and 3 there are other close intermolecular Ag...H(3)C contacts in the solid state, arising from proximate carborane anions in the crystal lattice. Addition of methyl groups to the periphery of the phosphine ligand (complex 4) switches off the majority of these interactions, leaving essentially a single cage interacting with the cationic silver-phosphine fragment through three CH(3) groups. In solution (CD(2)Cl(2)) Ag...H(3)C contacts remain, as evidenced by both the downfield chemical shift change and the significant line-broadening observed for the cage methyl signals. These studies also show that the metal fragment is fluxional over the surface of the cage. The Ag...H(3)C interactions in solution may be switched off by addition of a stronger Lewis base than [closo-1-H CB(11)Me(11)](-). Thus, addition of [NBu(4)][closo-1-H-CB(11)H(5)Br(6)] to 2 affords (Ph(3)P)Ag[closo-1-H-CB(11)H(5)Br(6)], while adding Et(2)O or PPh(3) affords the well-separated ion-pairs [(Ph(3)P)(L)Ag][closo-1-H-CB(11)Me(11)] (L = OEt(2) 5, PPh(3) 6,) both of which have been crystallographically characterized. DFT calculations on 2 (at the B3LYP/DZVP level) show small energy differences between the possible coordination isomers of this compound, with the favored geometry being one in which the [(Ph(3)P)Ag](+) fragment interacts with three of the [BCH(3)] vertices on the lower surface of the cage, similar to the experimentally observed structure of 4. PMID- 14759210 TI - A calix[4]arene carceplex with four Rh2 4+ fasteners. AB - It is shown that a spheroidal carceplex can be assembled by linking two bowl shaped calix[4]arenes via four dimetal units, (DAniF)(2)Rh(2) (DAniF = N,N'-di-p anisylformamidinate), with a molecule (diethyl ether) or a cation (tetraethylammonium ion) trapped inside. The tetraethylammonium carceplex, 1b, has been characterized by X-ray crystallography, (1)H NMR, IR, and mass spectrometry. The tetraethylammonium ion fits snugly in the interior of the spheroidal carceplex. A two-fold axis of the tetrahedral cation coincides with the idealized four-fold axis of the cage, and the cation is disordered over two rotational orientations. Only one axial position on each dirhodium unit is occupied by a ligand, CH(3)CN or H(2)O. The carceplex is very stable, and the axial ligands can be exchanged in single crystals without disrupting the crystallinity of the samples. In this way, a red crystal of the complex with all axial positions occupied by acetonitrile can be transformed to a green crystal of the complex with two axial positions having acetonitrile and the other two having water by simply putting the crystal in contact with distilled water. The calix[4]arene used to make the carciplex structure is 25,26,27,28-tetra-n propoxycalix[4]arene-5,11,17,23-tetracarboxylic acid. By employing 25,26,27,28 tetrapropoxy-5,17-dibromo-calix[4]arene-11,23-dicarboxylic acid, two 1:1 dimetal:calixarene compounds have also been made and characterized: 2, cis Rh(2)(DAniF)(2)(calix)(CH(3)OH), and 3, cis-Mo(2)(DAniF)(2)(calix). The Rh-Rh distances in 1b are in the range of 2.410(2)-2.428(2) A, that in 2 is 2.4383(4) A, and the Mo-Mo distance in 3 is 2.0931(4) A. PMID- 14759211 TI - Encapsulation of transition metal catalysts by ligand-template directed assembly. AB - Encapsulated transition metal catalysts are presented that are formed by templated self-assembly processes of simple building blocks such as porphyrins and pyridylphosphine and phosphite ligands, using selective metal-ligand interactions. These ligand assemblies coordinate to transition metals, leading to a new class of transition metal catalysts. The assembled catalyst systems were characterized using NMR and UV-vis spectroscopy and were identified under catalytic conditions using high-pressure infrared spectroscopy. Tris-3 pyridylphosphine binds three mesophenyl zinc(II) porphyrin units and consequently forms an assembly with the phosphorus donor atom completely encapsulated. The encapsulated phosphines lead exclusively to monoligated transition metal complexes, and in the rhodium-catalyzed hydroformylation of 1-octene the encapsulation of the catalysts resulted in a 10-fold increase in activity. In addition, the branched aldehyde was formed preferentially (l/b = 0.6), a selectivity that is highly unusual for this substrate, which is attributed to the encapsulation of the transition metal catalysts. An encapsulated rhodium catalyst based on ruthenium(II) porphyrins and tris-meta-pyridyl phosphine resulted in an even larger selectivity for the branched product (l/b = 0.4). These encapsulated catalysts can be prepared easily, and various template ligands and porphyrins, such as tris-3-pyridyl phosphite and ruthenium(II) porphyrins, have been explored, leading to catalysts with different properties. PMID- 14759212 TI - The role of weakly bound on-top oxygen in the catalytic CO oxidation reaction over RuO2(110). AB - RuO(2)-based catalysts are much more active in the oxidation of CO than related metallic Ru catalysts. This high catalytic activity (or low activation barrier) is attributed to the weak oxygen surface bonding of bridging O atoms on RuO(2)(110) in comparison with the strongly chemisorbed oxygen on Ru(0001). Since the RuO(2)(110) surface is able to stabilize an even more weakly bound on-top oxygen species, one would anticipate that the catalytic activity will increase further under oxidizing conditions. We will show that this view is far too simple to explain our temperature-programmed reaction experiments, employing isotope labeling of the potentially active surface oxygen species on RuO(2)(110). Rather, both surface O species on RuO(2)(110) reveal similar activities in oxidizing CO. PMID- 14759213 TI - New model for a theoretical density functional theory investigation of the mechanism of the carbonic anhydrase: how does the internal bicarbonate rearrangement occur? AB - A theoretical density functional theory (DFT, B3LYP) investigation has been carried out on the catalytic cycle of the carbonic anhydrase. A model system including the Glu106 and Thr199 residues and the "deep" water molecule has been used. It has been found that the nucleophilic attack of the zinc-bound OH on the CO(2) molecule has a negligible barrier (only 1.2 kcal mol(-1)). This small value is due to a hydrogen-bond network involving Glu106, Thr199, and the deep water molecule. The two usually proposed mechanisms for the internal bicarbonate rearrangement have been carefully examined. In the presence of the two Glu106 and Thr199 residues, the direct proton transfer (Lipscomb mechanism) is a two-step process, which proceeds via a proton relay network characterized by two activation barriers of 4.4 and 9.0 kcal mol(-1). This pathway can effectively compete with a rotational mechanism (Lindskog mechanism), which has a barrier of 13.2 kcal mol(-1). The fast proton transfer found here is basically due to the effect of the Glu106 residue, which stabilizes an intermediate situation where the Glu106 fragment is protonated. In the absence of Glu106, the barrier for the proton transfer is much larger (32.3 kcal mol(-1)) and the Lindskog mechanism becomes favored. PMID- 14759214 TI - Structure dependence of NO adsorption and dissociation on platinum surfaces. AB - The influence of surface structure on NO chemisorption and dissociation on Pt[100]-(1x1), Pt[211], and Pt[410] has been studied using density functional theory slab calculations with the generalized gradient corrections. The presence of steps on Pt[211] strengthens the NO-surface chemisorption bond, but the barrier for NO dissociation remains high. On the other hand, the steps on Pt[410] help to stabilize the N and O adatoms that form upon dissociation and the transition state. The calculated barrier of 80.2 kJ/mol on Pt[410] is in good agreement with experiment. These results show that both the presence of steps and the nature of the steps are important to activate NO. An ensemble of square arranged Pt atoms has been identified as an important feature in activating the N O bond. PMID- 14759215 TI - Affinity transformation from hydrophilicity to hydrophobicity of water molecules on the basis of adsorption of water in graphitic nanopores. AB - The interaction of water with hydrophobic surfaces is quite important in a variety of chemical and biochemical phenomena. The coexistence of water and oil can be realized by introduction of surfactants. In the case of water vapor adsorption on graphitic nanopores, plenty of water can be adsorbed in graphitic nanopores without surfactants, although the graphitic surface is not hydrophilic. Why are water molecules adsorbed in hydrophobic nanopores remarkably? This work can give an explicit insight to water adsorption in hydrophobic graphite nanopores using experimental and theoretical approaches. Water molecules are associated with each other to form the cluster of 1 nm in size, leading to a significant stabilization of the cluster in the graphitic nanopores. This mechanism can be widely applied to interfacial phenomena relating to coexistence of water and nanostructural materials of hydrophobicity. PMID- 14759216 TI - An investigation of the mechanisms of electronic sensing of protein adsorption on carbon nanotube devices. AB - It has been reported that protein adsorption on single-walled carbon nanotube field effect transistors (FETs) leads to appreciable changes in the electrical conductance of the devices, a phenomenon that can be exploited for label-free detection of biomolecules with a high potential for miniaturization. This work presents an elucidation of the electronic biosensing mechanisms with a newly developed microarray of nanotube "micromat" sensors. Chemical functionalization schemes are devised to block selected components of the devices from protein adsorption, self-assembled monolayers (SAMs) of methoxy(poly(ethylene glycol))thiol (mPEG-SH) on the metal electrodes (Au, Pd) and PEG-containing surfactants on the nanotubes. Extensive characterization reveals that electronic effects occurring at the metal-nanotube contacts due to protein adsorption constitute a more significant contribution to the electronic biosensing signal than adsorption solely along the exposed lengths of the nanotubes. PMID- 14759217 TI - Kinetic inhibitor of hydrate crystallization. AB - We present the results of a combined theoretical/experimental study into a new class of kinetic inhibitor of gas hydrate formation. The inhibitors are based on quaternary ammonium zwitterions, and were identified from a computational screen. Molecular dynamics simulations were used to characterize the effect of the inhibitor on the interface between a type II hydrate and natural gas. These simulations show that the inhibitor is bifunctional, with the hydrophobic end being compatible with the water structure present at the hydrate interface, while the negatively charged functional group promotes a long ranged water structure that is inconsistent with the hydrate phase; the sulfonate-induced structure was found to propagate strongly over several solvation shells. The compound was subsequently synthesized and used in an experimental study of both THF and ethane hydrate formation, and was shown to have an activity that was comparable with an existing commercial kinetic inhibitor: PVP. PMID- 14759218 TI - Beta-cyclodextrin host-guest complexes probed under thermodynamic equilibrium: thermodynamics and AFM force spectroscopy. AB - The rupture forces of individual host-guest complexes between beta-cyclodextrin (beta-CD) heptathioether monolayers on Au(111) and several surface-confined guests were measured in aqueous medium by single molecule force spectroscopy using an atomic force microscope. Anilyl, toluidyl, tert-butylphenyl, and adamantylthiols (0.2-1%) were immobilized in mixed monolayers with 2 mercaptoethanol on gold-coated AFM tips. For all guests and for all surface coverages, the force-displacement curves measured between the functionalized tips and monolayers of beta-CD exhibited single, as well as multiple, pull-off events. The histograms of the pull-off forces showed several maxima at equidistant forces, with force quanta characteristic for each guest of 39 +/- 15, 45 +/- 15, 89 +/- 15, and 102 +/- 15 pN, respectively. These force quanta were independent of the loading rate, indicating that, because of the fast complexation/decomplexation kinetics, the rupture forces were probed under thermodynamic equilibrium. The force values followed the same trend as the free binding energy Delta G degrees measured for model guest compounds in solution or on beta-CD monolayers, as determined by microcalorimetry and surface plasmon resonance measurements, respectively. A descriptive model was developed to correlate quantitatively the pull-off force values with the Delta G degrees of the complexes, based on the evaluation of the energy potential landscape of tip surface interaction. PMID- 14759219 TI - Sympathetic neural hyperactivity and its normalization following unstable angina and acute myocardial infarction. AB - Impaired autonomic function occurs after AMI (acute myocardial infarction) and UA (unstable angina), which may be important prognostically. However, the pattern of sympathetic nerve hyperactivity has been investigated only after AMI. We aimed to quantify central sympathetic output to the periphery in patients with UA, investigate its progress over time relative to that after uncomplicated AMI and to explore the mechanisms involved. Muscle sympathetic nerve activity (MSNA) assessed from multiunit discharges and from single units (s-MSNA) was obtained in matched patients with UA ( n =9), AMI ( n =14) and stable CAD (coronary artery disease, n =11), patients with chest pain in which AMI was excluded (NMI, n =9) and normal controls (NCs, n =14). Measurements were obtained 2-4 days after UA or AMI, and repeated at 3 monthly intervals until they returned to normal levels. The respective MSNA and s-MSNA early after UA (72+/-4.0 bursts/100 beats and 78+/ 4.2 impulses/100 beats respectively) were less than those after AMI (83+/-4.4 bursts/100 beats and 93+/-5.5 impulses/100 beats respectively). Relative to the control groups of NCs (51+/-2.7 bursts/100 beats and 58+/-3.4 impulses/100 beats respectively) and patients with CAD (54+/-3.7 bursts/100 beats and 58+/-3.9 impulses/100 beats respectively) and NMI (52+/-4.5 bursts/100 beats and 59+/-4.9 impulses/100 beats respectively), values returned to normal after 6 months in UA (55+/-5.0 bursts/100 beats and 62+/-5.5 impulses/100 beats respectively) and 9 months after AMI (60+/-3.8 bursts/100 beats and 66+/-4.2 impulses/100 beats respectively). In conclusion, both UA and AMI result in sympathetic hyper activity, although this is of smaller magnitude in UA and is less protracted than in AMI. It is suggested that this hyperactivity is related to the degree of left ventricular dysfunction and reflexes. PMID- 14759220 TI - Glycogenesis and glucose oxidation during an intravenous glucose tolerance test in man. AB - The quantity of deuterated glucose customarily given in labelled IVGTTs (intravenous glucose tolerance tests) changes the isotopic composition of the subject's body water enough to be detected by mass spectrometric techniques. Glucose undergoing direct glycogenesis does not contribute label to the body water pool, and isotope incorporated into it must have come from glucose that has either been oxidized or undergone indirect glycogenesis. By subtracting the amount of label found in body water from the total amount of glucose utilized, as calculated from the minimal model of glucose disappearance, it should be possible to study the partitioning of the dose given between direct glycogenesis in skeletal muscle and other metabolic pathways. To establish these principles, we used isotope ratio MS to determine body water composition in groups of healthy ( n =7; mean weight, 76 kg; fasting plasma glucose and insulin, 5.1 mmol and 40 pmol respectively) and Type II diabetic ( n =5; mean weight, 84 kg; fasting plasma glucose and insulin, 6.2 mmol and 75 pmol respectively) subjects undergoing IVGTTs. It was found that, for healthy subjects, 31% of the dose given was utilized in direct glycogenesis and this was decreased to 15% in diabetes. Defects in muscle glycogen synthesis in diabetes of the same order are well known from magnetic resonance studies. We conclude that measurement of label incorporation into body water is potentially useful for investigation of the metabolism of a glucose load in vivo during an IVGTT. PMID- 14759222 TI - Mutation of leucine-92 selectively reduces the apparent affinity of inosine, guanosine, NBMPR [S6-(4-nitrobenzyl)-mercaptopurine riboside] and dilazep for the human equilibrative nucleoside transporter, hENT1. AB - We developed a yeast-based assay for selection of hENT1 (human equilibrative nucleoside transporter 1) mutants that have altered affinity for hENT1 inhibitors and substrates. In this assay, expression of hENT1 in a yeast strain deficient in adenine biosynthesis (ade2) permits yeast growth on a plate lacking adenine but containing adenosine, a hENT1 substrate. This growth was prevented when inhibitors of hENT1 [e.g. NBMPR [S6-(4-nitrobenzyl)-mercaptopurine riboside], dilazep or dipyridamole] were included in the media. To identify hENT1 mutants resistant to inhibition by these compounds, hENT1 was randomly mutagenized and introduced into this strain. Mutation(s) that allowed growth of yeast cells in the presence of these inhibitors were then identified and characterized. Mutants harbouring amino acid changes at Leu92 exhibited resistance to NBMPR and dilazep but not dipyridamole. The IC50 values of NBMPR and dilazep for [3H]adenosine transport by one of these mutants L92Q (Leu92-->Gln) were approx. 200- and 4-fold greater when compared with the value for the wild-type hENT1, whereas that for dipyridamole remained unchanged. Additionally, when compared with the wild-type transporter, [3H]adenosine transport by L92Q transporter was significantly resistant to inhibition by inosine and guanosine but not by adenosine or pyrimidines. The Km value for inosine transport was approx. 4-fold greater for the L92Q mutant (260+/-16 mM) when compared with the wild-type transporter (65+/ 7.8 mM). We have identified for the first time an amino acid residue (Leu92) of hENT1 that, when mutated, selectively alters the affinity of hENT1 to transport the nucleosides inosine and guanosine and its sensitivity to the inhibitors NBMPR and dilazep. PMID- 14759223 TI - Comparative binding character of two general anaesthetics for sites on human serum albumin. AB - Propofol and halothane are clinically used general anaesthetics, which are transported primarily by HSA (human serum albumin) in the blood. Binding characteristics are therefore of interest for both the pharmacokinetics and pharmacodynamics of these drugs. We characterized anaesthetic-HSA interactions in solution using elution chromatography, ITC (isothermal titration calorimetry), hydrogen-exchange experiments and geometric analyses of high-resolution structures. Binding affinity of propofol to HSA was determined to have a K(d) of 65 microM and a stoichiometry of approx. 2, whereas the binding of halothane to HSA showed a K(d) of 1.6 mM and a stoichiometry of approx. 7. Anaesthetic-HSA interactions are exothermic, with propofol having a larger negative enthalpy change relative to halothane. Hydrogen-exchange studies in isolated recombinant domains of HSA showed that propofol-binding sites are primarily found in domain III, whereas halothane sites are more widely distributed. Both location and stoichiometry from these solution studies agree with data derived from X-ray crystal-structure studies, and further analyses of the architecture of sites from these structures suggested that greater hydrophobic contacts, van der Waals interactions and hydrogen-bond formation account for the stronger binding of propofol as compared with the less potent anaesthetic, halothane. PMID- 14759224 TI - Nucleotides and transported substrates modulate different steps of the ATPase catalytic cycle of MRP1 multidrug transporter. AB - The human ABC (ATP-binding cassette) transporter MRP1 (human multidrug-resistance associated protein 1; ABCC1) is involved in the cellular extrusion of conjugated metabolites and causes multidrug resistance in tumour cells. The transport of substrate molecules by ABC proteins is energized by ATP hydrolysis, performed by two co-operating ABC units. Orthovanadate (Vi), a non-covalent inhibitor of the ABC ATPases, was found to catalyse a photo-oxidative cleavage of various ATP binding proteins. In the present study, we have identified three Vi-cleavage sites within MRP1, and found that the cleavage reactions were variably modulated by the presence of nucleotides and by transported substrates. We concluded that Vi cleavage of MRP1 at Site I detects conformational changes due to the binding of MgATP. In contrast, Site II could be identified as part of the substrate modulated catalytic cycle, probably containing an MRP1.MgADP.Vi transition-state like complex. Cleavage at Site III was modulated by both the binding and hydrolysis of MgATP, in a biphasic pattern, which was also affected by the presence of transported substrates. We detected two different allosteric effects and found that they control two consecutive steps of the MRP1 ATPase catalytic cycle. Nucleotide binding to the low-affinity site accelerated the formation of the pre-hydrolytic intermediate in the other catalytic centre. Interaction of the transporter with its transported substrates stimulated a later reaction of the hydrolytic cycle, the formation of the post-hydrolytic intermediate, which could be detected in both catalytic sites by the experimental strategy used. PMID- 14759225 TI - Control of the CDPethanolamine pathway in mammalian cells: effect of CTP:phosphoethanolamine cytidylyltransferase overexpression and the amount of intracellular diacylglycerol. AB - For an insight regarding the control of PtdEtn (phosphatidylethanolamine) synthesis via the CDPethanolamine pathway, rat liver cDNA encoding ECT (CTP:phosphoethanolamine cytidylyltransferase) was transiently or stably transfected in Chinese-hamster ovary cells and a rat liver-derived cell line (McA RH7777), resulting in a maximum of 26- and 4-fold increase in specific activity of ECT respectively. However, no effect of ECT overexpression on the rate of [3H]ethanolamine incorporation into PtdEtn was detected in both cell lines. This was explored further in cells overexpressing four times ECT activity (McA-ECT1). The rate of PtdEtn breakdown and PtdEtn mass were not changed in McA-ECT1 cells in comparison with control-transfected cells. Instead, an accumulation of CDPethanolamine (label and mass) was observed, suggesting that in McA-ECT1 cells the ethanolaminephosphotransferase-catalysed reaction became rate-limiting. However, overexpression of the human choline/ethanolaminephosphotransferase in McA-ECT1 and control-transfected cells had no effect on PtdEtn synthesis. To investigate whether the availability of DAG (diacylglycerol) limited PtdEtn synthesis in these cells, intracellular DAG levels were increased using PMA or phospholipase C. Exposure of cells to PMA or phospholipase C stimulated PtdEtn synthesis and this effect was much more pronounced in McA-ECT1 than in control transfected cells. In line with this, the DAG produced after PMA exposure was consumed more rapidly in McA-ECT1 cells and the CDPethanolamine level decreased accordingly. In conclusion, our results suggest that the supply of CDPethanolamine, via the expression level of ECT, is an important factor governing the rate of PtdEtn biosynthesis in mammalian cells, under the condition that the amount of DAG is not limiting. PMID- 14759227 TI - Introduction: standards of antibacterial performance. AB - The development and clinical use of antimicrobial agents continue to evolve in line with new science, understanding and needs. While antimicrobial resistance remains an important determinant for drug development and therapeutic choice, pharmacokinetic and pharmacodynamic parameters are having an ever-increasing importance in defining performance targets for new and established agents. Recently licensed new therapies are largely directed at serious hospital associated Gram-positive infections, whereas in the community, therapeutic choice remains dependent on well-established agents from limited classes of antimicrobials. In order to maximise the benefits from such agents, it is appropriate that dosage regimens and antibacterial choices be reviewed in the light of new knowledge, particularly in the area of pharmacokinetics and pharmacodynamics. Antimicrobial resistance continues to evolve, notably within respiratory pathogens, therefore steps must be taken to maintain optimum therapeutic outcomes and also limit the development and spread of resistant strains. Whilst changes in patient and physician attitudes and behaviour towards better quality prescribing are important, new agents must also be developed to provide adequate coverage for resistant pathogens. Development times for novel agents and classes of antimicrobials are long, with uncertain safety profiles and chances of success. Thus, the development of new formulations of existing agents, designed to overcome current resistance patterns, constitutes a potentially important additional strategy towards appropriate prescribing. PMID- 14759228 TI - Elements of design: the knowledge on which we build. AB - The time the free drug serum concentration of antibiotic remains above the pathogen MIC (T > MIC) determines bacteriological efficacy and emergence or selection of resistance for penicillin and amoxicillin with or without clavulanate. Multiple studies in animal and in-vitro models now support this conclusion. The size of the T > MIC (the pharmacokinetic/-dynamic target) is > 40 50% to maximise antibacterial effect and pathogen eradication for Streptococcus pneumoniae and probably also Haemophilus influenzae. The size of the T > MIC for optimal antibacterial effect is changed by host immune status but not by bacterial inoculum or mechanism of resistance. There is good animal evidence to support the prediction that, as long as the target T > MIC is achieved, strains of S. pneumoniae with amoxicillin MICs of 0.016 mg/L will respond to amoxicillin in the same way as those with MICs of 1-2 mg/L. Emergence of resistance to amoxicillin/clavulanate in S. pneumoniae is related to low T > MIC (< 20%) and also to the degree of population heterogeneity to amoxicillin. Selection of resistant strains of S. pneumoniae is also related to T > MIC. Monte Carlo simulations based on the pharmacokinetics of amoxicillin with or without clavulanate in humans are needed to best predict the likely efficacy of different amoxicillin dosing regimens. This approach adequately allows the considerable pharmacokinetic variability in amoxicillin handling by infected patients to be accounted for as well as differences in pathogen beta-lactam susceptibility. PMID- 14759226 TI - Intracellular proteoglycans. AB - Proteoglycans (PGs) are proteins with glycosaminoglycan chains, are ubiquitously expressed and have a wide range of functions. PGs in the extracellular matrix and on the cell surface have been the subject of extensive structural and functional studies. Less attention has so far been given to PGs located in intracellular compartments, although several reports suggest that these have biological functions in storage granules, the nucleus and other intracellular organelles. The purpose of this review is, therefore, to present some of these studies and to discuss possible functions linked to PGs located in different intracellular compartments. Reference will be made to publications relevant for the topics we present. It is beyond the scope of this review to cover all publications on PGs in intracellular locations. PMID- 14759229 TI - Proof of concept: performance testing in models. AB - Pharmacokinetic (PK) and pharmacodynamic (PD) principles that predict antimicrobial efficacy can be used to set targets for antimicrobial design and optimisation. Although current formulations of amoxicillin and amoxicillin/clavulanate have retained their efficacy against many, but not all, penicillin-nonsusceptible Streptococcus pneumoniae, additional coverage is required to address the growing problem of drug-resistant strains. Accordingly, two new oral formulations of amoxicillin/clavulanate, a paediatric formulation at 90/6.4 mg/kg/day and a pharmacokinetically enhanced formulation at 2000/125 mg twice daily for adults, were designed using PK/PD principles. These principles indicate that for amoxicillin and amoxicillin/clavulanate, a time above MIC of 35 40% of the dosing interval is predictive of high bacterial efficacy. In line with PK/PD predictions, simulation of human pharmacokinetics in in-vitro kinetic models and in a rat model of pneumonia, amoxicillin/clavulanate 2000/125 mg twice daily was highly effective against S. pneumoniae strains with amoxicillin MICs of 4 or 8 mg/L. Against strains with amoxicillin MICs of 4 mg/L, amoxicillin/clavulanate 2000/125 mg twice daily was significantly more effective than the conventional 875/125 mg twice daily formulation, azithromycin and levofloxacin, even though all levofloxacin MICs were < or = 1 mg/L. Following infection with S. pneumoniae strains with amoxicillin MICs of 8 mg/L, the amoxicillin/clavulanate 2000/125 mg twice daily formulation was more effective than the conventional amoxicillin/clavulanate formulations of 875/125 mg twice daily and three times daily and 1000/125 mg three times daily, and had similar or better efficacy than azithromycin and levofloxacin, depending on the strain. These data indicate the potential benefit of therapy with amoxicillin/clavulanate 2000/125 mg twice daily compared with conventional formulations and other marketed antimicrobials in the treatment of respiratory tract infection. PMID- 14759230 TI - Building in efficacy: developing solutions to combat drug-resistant S. pneumoniae. AB - The development of our understanding of the pharmacokinetic (PK) and pharmacodynamic (PD) principles that determine antimicrobial efficacy has advanced substantially over the last 10 years. We are now in a position to use PK/PD principles to set targets for antimicrobial design and optimisation so that we can predict eradication of specific pathogens or resistant variants when agents are used clinically. Optimisation of PK/PD parameters to enable the treatment of resistant pathogens with oral agents may not be possible with many current agents, such as some cephalosporins, macrolides and fluoroquinolones. Aminopenicillins, however, such as amoxicillin, have linear PK and have a good safety profile even at high doses. The new pharmacokinetically enhanced oral formulation of amoxicillin/clavulanate, 2000/125 mg twice daily, was designed using PK/PD principles to be able to eradicate Streptococcus pneumoniae with amoxicillin MICs of up to and including 4 mg/L, which includes most penicillin resistant isolates. For amoxicillin and amoxicillin/clavulanate, a time above MIC (T > MIC) of 35-40% of the dosing interval (based on blood levels) is predictive of high bacteriological efficacy. This target was met by the design of a unique bilayer tablet incorporating 437.5 mg of sustained-release sodium amoxicillin in one layer plus 562.5 mg of immediate-release amoxicillin trihydrate and 62.5 mg of clavulanate potassium in the second layer, with two tablets administered for each dose. This unique design extends the bacterial killing time by increasing the T > MIC to 49% of the dosing interval against pathogens with MICs of 4 mg/L, and 60% of the dosing interval against pathogens with MICs of 2 mg/L. Based on these results, this new amoxicillin/clavulanate formulation should be highly effective in treating respiratory tract infections due to drug-resistant S. pneumoniae as well as beta-lactamase-producing pathogens, such as Haemophilus influenzae and Moraxella catarrhalis. PMID- 14759231 TI - Performance in practice: bacteriological efficacy in patients with drug-resistant S. pneumoniae. AB - Using pharmacokinetic/pharmacodynamic principles, pharmacokinetically enhanced amoxicillin/clavulanate 2000/125 mg twice daily was designed to provide adequate levels of amoxicillin over the 12-h dosing interval to eradicate penicillin resistant Streptococcus pneumoniae (PRSP, penicillin MICs > or = 2 mg/L) with amoxicillin MICs of at least 4 mg/L. The clinical efficacy of amoxicillin/clavulanate 2000/125 mg was evaluated in patients with respiratory tract infections caused by S. pneumoniae, including isolates with elevated penicillin (2-8 mg/L) MICs. Data from 10 clinical studies were combined: seven randomised (1:1), double-blind, controlled trials (efficacy intent-to-treat [ITT]N = 3376): amoxicillin/clavulanate 2000/125 mg twice daily vs. levofloxacin 500 mg once daily in acute bacterial sinusitis (ABS); levofloxacin 500 mg once daily in acute exacerbations of chronic bronchitis (AECB); clarithromycin 500 mg twice daily in AECB; amoxicillin/clavulanate 875/125 mg twice daily/three times daily and 1000/125 mg three times daily in community-acquired pneumonia (CAP) and three noncomparative studies (efficacy ITT N = 3024): two in ABS, one in CAP. The bacteriological per-protocol (PP) population at follow up (days 14-39) comprised 1295 patients for amoxicillin/clavulanate 2000/125 mg and 241 for comparators. With amoxicillin/clavulanate 2000/125 mg at follow-up, outcome was successful (clinical success and eradication/presumed eradication) in 85/90 (94.4%) patients with S. pneumoniae in comparative studies and 421/445 (94.6%) in noncomparative studies, and with comparators 58/70 (82.9%) were successes. In the amoxicillin/clavulanate 2000/125 mg group at follow up, 52/552 S. pneumoniae isolates were resistant to penicillin. At follow up, 50/52 (96.2%) patients with PRSP were successes, including 6/7 with amoxicillin MICs of 4 mg/L and 7/8 with amoxicillin MICs of 8 mg/L. Success rates for amoxicillin/clavulanate 2000/125 mg against PRSP were similar for CAP (96.0%[24/25]), AECB (100%[3/3]) and ABS (95.8%[23/24]). There were six PRSP isolates in the comparator group (two isolates were from one patient), and three of five patients in this group were successes. In conclusion, amoxicillin/clavulanate 2000/125 mg demonstrated combined clinical/bacteriological success against 50/52 patients with PRSP, including 13/15 strains with amoxicillin MICs of 4-8 mg/L. These results for the pharmacokinetic-enhanced formulation of amoxicillin/clavulanate 2000/125 mg are in line with the high efficacy against PRSP predicted using pharmacokinetic/pharmacodynamic parameters. PMID- 14759233 TI - Schistosomiasis as an unusual cause of appendicitis. AB - Millions of people originating from tropical areas now live outside the country of their birth. As a consequence, the number of cases of diseases imported from the tropics and being seen by European physicians in immigrants is growing. As an example of such diseases, schistosomal appendicitis is a specific trait of infection with Schistosoma haematobium and is an uncommon cause of appendicitis in non-endemic areas. Treatment requires anti-schistomal medication in addition to surgery. Physicians, including surgeons, need to be aware of the possibility of seeing atypical presentations of parasitic diseases in immigrant patients. PMID- 14759234 TI - Multilocus sequence typing and the evolution of methicillin-resistant Staphylococcus aureus. AB - The prevalence of methicillin-resistant Staphylococcus aureus (MRSA) in many countries is increasing and, in hospitals in some areas, more than half of all S. aureus disease isolates are MRSA. MRSA strains are becoming increasingly multiresistant, and have recently developed resistance to vancomycin, used successfully to treat MRSA for more than 30 years. This review summarises recent studies that have elucidated the evolutionary history of MRSA. The first MRSA isolate evolved from a sensitive, epidemic strain prevalent in Europe, and its progeny-the first MRSA clone-quickly spread to other continents. Analyses of epidemic MRSA isolates from hospitals in different countries by molecular methods, including multilocus sequence typing (MLST) and DNA microarray analysis, reveal that MRSA strains have evolved separately within five distinct epidemic, sensitive lineages. However, resistance has been transferred to S. aureus on many more than five occasions, as some lineages have acquired different structural types of the element carrying the methicillin resistance gene. The emergence of MRSA as a community pathogen has been noted in several countries, and MLST and SCCmec typing have been used to demonstrate that community-acquired MRSA strains are typically related only distantly to hospital MRSA strains, and thus represent novel acquisitions of SCCmec. PMID- 14759235 TI - Actinobacillus actinomycetemcomitans endocarditis. AB - Among the bacteria of the HACEK group, Actinobacillus actinomycetemcomitans is the organism involved most commonly in infective endocarditis. However, the epidemiological and clinical features specifically associated with this species have not been evaluated adequately. Three patients with infective endocarditis caused by A. actinomycetemcomitans seen at the Hospital La Timone (Marseille, France) between 1994 and 2001 are reported. Of 99 cases in the literature, 75% of patients had previous heart disease before infective endocarditis, the portal of entry of which was usually the oral cavity. Among the total of 102 cases, 27 had prosthetic valves. Intermittent fever was observed in all cases, and weight loss and peripheral signs of endocarditis were noteworthy in this study. Anaemia and microscopic haematuria were frequently noted. The disease is insidious, with a mean duration of symptoms of 13 weeks before diagnosis, as confirmed by blood cultures incubated for > 5 days. The aortic valve is most commonly involved, and echocardiographic findings were non-specific. Complications occurred in 63% of patients, with emboli being the most common. The surgery rate was 23.5%. The overall mortality rate was 18%. Of the cases, 76.5% were cured with antibiotics alone, including a simple third-generation cephalosporin or a combination of ampicillin and an aminoglycoside. An antibiotic therapy duration of at least 4 weeks is recommended. Surgical therapy is usually required for haemodynamic reasons. Prophylaxis of A. actinomycetemcomitans endocarditis relies on antibiotic therapy for all cardiac patients at risk before dental procedures. Among 17 patients undergoing dental manipulations, only eight received amoxycillin before the procedure, demonstrating that prophylaxis is far from being systematically prescribed. In conclusion, A. actinomycetemcomitans endocarditis should be highly suspected in patients with previous cardiac disease and for whom symptoms have evolved over a number of weeks or even months. PMID- 14759236 TI - Effect of polymorphisms on the replicative capacity of protease inhibitor resistant HIV-1 variants under drug pressure. AB - The role of drug pressure on the replicative capacity of protease inhibitor resistant HIV-1 variants and the contribution of a common amino-acid polymorphism in the protease gene (L63P) to this process were investigated. Using HIV-1 variants resistant to the protease inhibitors saquinavir (G48V/L90M) or indinavir (A71V/V82T/I84V), viral replication was studied in the presence or absence of inhibitor and a mutation at position 63. The initial changes diminished enzyme function of the protease and reduced replicative capacity for both virus mutants. Addition of the respective inhibitor blocked the wild-type, but was also able to delay the replication kinetics of either mutant, revealing the limits of resistance. Importantly, the polymorphic change L63P, although not conferring inhibitor resistance by itself, provided a significant replication benefit to both mutant viruses, particularly under drug pressure, and may reveal a far reaching compensating power of polymorphic changes. This may drive evolution and the directed selection of protease inhibitor-resistant HIV-1 variants, a finding with significant clinical and diagnostic implications. PMID- 14759237 TI - Peptide insertions in reverse transcriptase pol gene of human immunodeficiency virus type 1 as a rare cause of persistent antiretroviral therapeutic failure. AB - Peptide insertions in codons 67-71 of the reverse transcriptase (RT) pol gene were detected in 11 (2.7%) of 414 genotypic analyses performed in a hospital cohort of 2900 outpatients with human immunodeficiency virus type 1 (HIV-1) infection. The duration of antiretroviral treatment (bi- or tri-therapy) before the detection of insertions ranged from 12 to 60 months. Dipeptide insertions were detected in ten patients, of which the most frequent was serine-serine. A monopeptide insertion was diagnosed once. The amino-acid composition patterns of insertions varied with time in five of the 11 patients. Peptide insertions were always associated with various patterns of pre-existing or appearing resistance mutations in the RT pol gene to different antiretroviral drugs. Genotypic-guided treatment resulted in virological and immunological improvement in two patients. In contrast, the remaining patients did not respond to any of the various antiretroviral regimens prescribed. Furthermore, various patterns of resistance mutations developed to the prescribed antiretroviral drugs, with AIDS-related conditions leading to death in two patients. It was concluded that peptide insertion in this region of the HIV-1 RT pol gene constitutes a rare cause of persistent therapeutic failure, and that management of such patients remains challenging despite successive genotypic analyses aimed at detecting mutations conferring antiretroviral drug resistance. PMID- 14759238 TI - Carriage of Neisseria meningitidis by Greek children: risk factors and strain characteristics. AB - Oropharyngeal swabs were cultured from 554 children aged 2-19 years attending nurseries, primary schools and secondary schools in the central Athens area. A questionnaire was completed to identify risk factors for carriage. Susceptibility to antimicrobial agents was determined by Etest. The genetic relatedness of the strains was examined by pulsed-field gel electrophoresis (PFGE), and isolate serogrouping was performed by slide agglutination. Twenty-two (4%) children were carriers of Neisseria meningitidis; seven isolates belonged to serogroup C, and five to serogroup B. One isolate was resistant to co-trimoxazole, and five showed intermediate resistance to penicillin. DNA analysis of 16 isolates revealed six distinct PFGE patterns. Clusters with indistinguishable PFGE patterns were noted in the same school. More than one serogroup was included in the same clonal group. On multivariate logistic regression analysis, only age > 12 years remained independently associated with the carrier state (odds ratio, 7.96; 95% CI, 2.24 28.33; p < 0.001). Overall, the N. meningitidis carriage rate among Greek schoolchildren increased with age, and the predominant serogroups in the Athens region were groups C and B. These findings may have important implications for future immunisation strategies with conjugate vaccines. PMID- 14759239 TI - Molecular characterisation of an outbreak strain of multiresistant Salmonella enterica serovar Typhimurium DT104 in the UK. AB - A major national outbreak of multiresistant Salmonella enterica serovar Typhimurium definitive phage type 104 (MR DT104) occurred in England and Wales in the summer of 2000. Isolates of MR DT104 were characterised by antimicrobial resistance type (R-type), pulsed-field gel electrophoresis (PFGE), plasmid profiling and fluorescent amplified fragment length polymorphism (fAFLP) analysis. Results of R-type, PFGE and fAFLP showed that summer 2000 outbreak associated isolates were indistinguishable from most MR DT104 isolates collected in England and Wales during the 1980s and 1990s. However, outbreak-associated isolates all had an additional 2-MDa plasmid (PP D), and this distinct profile allowed outbreak cases to be distinguished from background MR DT104 infections, thereby facilitating the epidemiological investigation by improving the specificity of the case definition. The study demonstrated the highly clonal nature of MR DT104 and the importance of a hierarchical approach to molecular subtyping for outbreak investigations. PMID- 14759240 TI - Use of quantitative antibiogram analysis to determine the clonality of coagulase negative Staphylococcus species from blood culture. AB - Two phenotypic methods, quantitative antibiogram analysis and colony morphology, were compared to pulsed-field gel electrophoresis (PFGE) for distinguishing the clonality of coagulase-negative Staphylococcus (CNS) species. The results of these three methods were correlated with the patients' clinical findings for 23 episodes in which CNS species were isolated from two blood culture bottles within a 24-h period. Quantitative antibiogram and colony morphology at 24 h correlated with PFGE typing in 21 (91%) and 20 (87%) episodes, respectively. All episodes associated with CNS strains with identical PFGE patterns had quantitative antibiogram similarity coefficients < 10, whereas most episodes associated with strains with different PFGE patterns had quantitative antibiogram similarity coefficients >or= 17. The CNS isolate pairs were less likely to be associated with infection if the strains had different PFGE types or a quantitative antibiogram similarity coefficient >or= 17. Clinical microbiology laboratories should consider use of the quantitative antibiogram similarity coefficient to aid clinicians in distinguishing infection-associated CNS blood isolates from contaminants. PMID- 14759241 TI - Integrons as tools for epidemiological studies. AB - The integron content of Gram-negative strains implicated in three distinct episodes of suspected cross-infection among inpatients was investigated and compared with ribotyping. In the first episode, ribotyping identified a strain of Acinetobacter, isolated over a 3-month period, responsible for an outbreak associated with the use of mechanical ventilation in the intensive care unit (ICU). The second episode concerned simultaneous isolations of Pseudomonas aeruginosa and Serratia marcescens from 13 bronchoscopy patients. In these two episodes, results obtained by analysis of integron content and ribotyping were in agreement and correctly identified the epidemiologically related strains. In the third episode, isolates of Enterobacter cloacae were collected from patients in the neonatal ICU over a 3-month period. Although several isolates belonged to the same ribotype, cross-infection could not always be confirmed when the integron content was analysed. Integron detection can be considered a useful tool for studying molecular epidemiology in hospital environments, facilitating the quick detection of possible cross-infection cases, especially in critical wards such as the ICU. PMID- 14759242 TI - Ampicillin + sulbactam vs clindamycin +/- cephalosporin for the treatment of aspiration pneumonia and primary lung abscess. AB - Aspiration pneumonia, necrotising pneumonia and primary lung abscess are complications arising from the aspiration of infectious material from the oral cavity or stomach. There is limited information on optimal antibacterial therapeutic regimens. Patients with pulmonary infection following aspiration (n = 95) were included in a prospective, open, randomised, comparative multicentre trial to compare the safety, clinical and bacteriological efficacy of ampicillin + sulbactam vs. clindamycin +/- cephalosporin. Treated patients (n = 70) received sequential antibiotic therapy with either ampicillin + sulbactam (n = 37) or clindamycin (n = 33), with or without a second- or third-generation cephalosporin, administered until the complete resolution of clinical and radiological abnormalities. Definite or presumptive pathogens were isolated from 58 patients. Mean duration of therapy was 22.7 days for ampicillin + sulbactam and 24.1 days for clindamycin. In patients treated with ampicillin + sulbactam, the clinical response was 73.0% at the end of therapy and 67.5% 7-14 days after therapy. For clindamycin, the rates were 66.7% and 63.5%, respectively. Bacteriological response was similar in both treatment arms. Nine patients died (12.9%), with a Simplified Acute Physiology Score of > 30 points being the only significant predictive factor for therapeutic failure. Ampicillin + sulbactam and clindamycin +/- cephalosporin were both well-tolerated and proved equally effective in the treatment of aspiration pneumonia and lung abscess. PMID- 14759243 TI - The reproducibility of an enzyme-linked immunosorbent assay for detection of Chlamydia pneumoniae-specific antibodies. AB - With use of an enzyme-linked immunosorbent assay (ELISA), Chlamydia pneumoniae immunoglobulins were detected in a consecutive series of patients' sera (n = 122 for IgA and IgG; n = 138 for IgM). When the ELISA tests were repeated, the percentage disagreements were 12%, 16% and 10% for C. pneumoniae IgA, IgG and IgM, respectively. The reproducibility of the ELISA, expressed as kappa values, for IgA, IgG and IgM was 0.73, 0.60 and 0.53, respectively (p < 0.001). It was concluded that the ELISA had good reproducibility for detecting C. pneumoniae IgA, and moderately good reproducibility for detecting C. pneumoniae IgG and IgM. PMID- 14759244 TI - Similar inflammatory response in human whole blood to live Streptococcus pneumoniae of different serotypes. AB - Differences in inflammatory responses in human adult whole blood to live pneumococcal serotypes 3, 7F, 9V and 23F were investigated. Using flow cytometry and ELISA, oxidative burst, expression of activation markers CD11b/CD18, and in vitro production of tumour necrosis factor-alpha, interleukin-6 (IL-6) and interleukin-8 were measured. There was no significant difference between the serotypes regarding any of the variables investigated, although there was a trend towards higher concentrations of IL-6 induced by serotypes 9V and 23F. In the present experimental model, the serotypes of Streptococcus pneumoniae shown previously to cause different degrees of inflammation were found to cause a similar inflammatory response in human whole blood. PMID- 14759245 TI - Characterisation of invasive pneumococcal isolates in Catalan children up to 5 years of age, 1989-2000. AB - Ninety-six Streptococcus pneumoniae strains isolated between January 1989 and December 2000 from usually sterile sites of children aged < 5 years of age were included in the study. Resistance to penicillin (38.6% intermediate, 10.4% high level), cefotaxime (20.8%), tetracycline (41.7%), chloramphenicol (33.3%) and erythromycin (27.1%), as well as serogroup/type, were related to age and pathology. Strains from children aged < 2 years showed the highest penicillin resistance rate. Resistance to penicillin, tetracycline, chloramphenicol and erythromycin was the most common pattern (18.8% of strains). Most isolates (80.2%) belonged to serogroups/types included in the heptavalent conjugate vaccine. PMID- 14759246 TI - Serological investigation of the prevalence of anti-dengue IgM and IgG antibodies in Attapeu Province, South Laos. AB - The prevalence of dengue antibodies was determined in the Attapeu region of South Laos with 225 blood samples collected from mostly febrile patients during the rainy season August - October 2001. An IgM capture ELISA was positive for one (0.4%) sample, while 177 (79%) samples were positive in an indirect IgG ELISA. Of the positive IgG samples, 20 (11.3%) were also positive on blood slides for Plasmodium falciparum. Dengue fever seems to be widespread in this area, but clinical dengue diagnosis remains difficult, especially in the first days of illness when physicians have to discriminate between dengue and other febrile illnesses. PMID- 14759247 TI - Serum ferritin levels in West Nile encephalitis. AB - West Nile encephalitis (WNE) presents clinically as aseptic meningitis, meningoencephalitis, encephalitis, or acute flaccid paralysis. Non-specific laboratory findings, e.g., leukopenia and thrombocytopenia, accompany WNE. Lymphopenia is marked and prolonged with WNE. Three patients with WNE were found to have elevated serum ferritin levels. Severity seemed to be directly related to serum ferritin levels. Although preliminary, the results suggested that serum ferritin levels >or= 500 ng/mL (normal range 5-187 ng/mL) occur late with WNE, and not in a control group of patients with viral meningitis or encephalitis. PMID- 14759248 TI - Bad chemistry. AB - General chemistry courses haven't changed significantly in forty years. Because most basic chemistry students are premedical students, medical schools have enormous influence and could help us start all over again to create undergraduate chemistry education that works. PMID- 14759249 TI - Rewiring the transcriptional regulatory circuits of cells. AB - The molecular mechanisms that regulate gene expression can evolve either by changing the cis-acting DNA elements in promoters, or by replacing the trans acting regulatory proteins. New data from yeast species show that both processes can happen. PMID- 14759250 TI - The diversity of endothelial cells: a challenge for therapeutic angiogenesis. AB - Vascular endothelia comprise a diverse population of cells that specialize in response to genetic programs and environmental cues to take on distinct roles in different vessels, tissues, and organs, and in response to pathophysiological stresses. Characterization of endothelial-cell diversity will facilitate the development of novel, highly specific and safe therapies for many diseases. PMID- 14759251 TI - Spreading silence with Sid. AB - RNA interference (RNAi) has been shown to spread from cell to cell in plants and in Caenorhabditis elegans, but it does not spread in other organisms, such as Drosophila. A recent report demonstrates that a membrane channel, encoded by the gene sid-1, is responsible for the spreading of RNAi between cells. PMID- 14759253 TI - Functional genomics: strict tempo and hierarchical vocabularies. PMID- 14759252 TI - Adenosine deaminases acting on RNA (ADARs): RNA-editing enzymes. AB - Adenosine deaminases acting on RNA (ADARs) were discovered as a result of their ability extensively to deaminate adenosines in any long double-stranded RNA, converting them to inosines. Subsequently, ADARs were found to deaminate adenosines site-specifically within the coding sequences of transcripts encoding ion-channel subunits, increasing the diversity of these proteins in the central nervous system. ADARI is now known to be involved in defending the genome against viruses, and it may affect RNA interference. ADARs are found in animals but are not known in other organisms. It appears that ADARs evolved from a member of another family, adenosine deaminases acting on tRNAs (ADATs), by steps including fusion of two or more double-stranded-RNA binding domains to a common type of zinc-containing adenosine-deaminase domain. PMID- 14759254 TI - The post-genomic era for a select few. PMID- 14759255 TI - Making sense of lung-cancer gene-expression profiles. PMID- 14759256 TI - Mouse genomics gets the royal treatment. PMID- 14759258 TI - An unappreciated role for RNA surveillance. AB - BACKGROUND: Nonsense-mediated mRNA decay (NMD) is a eukaryotic mRNA surveillance mechanism that detects and degrades mRNAs with premature termination codons (PTC+ mRNAs). In mammals, a termination codon is recognized as premature if it lies more than about 50 nucleotides upstream of the final intron position. More than a third of reliably inferred alternative splicing events in humans have been shown to result in PTC+ mRNA isoforms. As the mechanistic details of NMD have only recently been elucidated, we hypothesized that many PTC+ isoforms may have been cloned, characterized and deposited in the public databases, even though they would be targeted for degradation in vivo. RESULTS: We analyzed the human alternative protein isoforms described in the SWISS-PROT database and found that 144 (5.8% of 2,483) isoform sequences amenable to analysis, from 107 (7.9% of 1,363) SWISS-PROT entries, derive from PTC+ mRNA. CONCLUSIONS: For several of the PTC+ isoforms we identified, existing experimental evidence can be reinterpreted and is consistent with the action of NMD to degrade the transcripts. Several genes with mRNA isoforms that we identified as PTC+--calpain-10, the CDC-like kinases (CLKs) and LARD--show how previous experimental results may be understood in light of NMD. PMID- 14759257 TI - A comprehensive evolutionary classification of proteins encoded in complete eukaryotic genomes. AB - BACKGROUND: Sequencing the genomes of multiple, taxonomically diverse eukaryotes enables in-depth comparative-genomic analysis which is expected to help in reconstructing ancestral eukaryotic genomes and major events in eukaryotic evolution and in making functional predictions for currently uncharacterized conserved genes. RESULTS: We examined functional and evolutionary patterns in the recently constructed set of 5,873 clusters of predicted orthologs (eukaryotic orthologous groups or KOGs) from seven eukaryotic genomes: Caenorhabditis elegans, Drosophila melanogaster, Homo sapiens, Arabidopsis thaliana, Saccharomyces cerevisiae, Schizosaccharomyces pombe and Encephalitozoon cuniculi. Conservation of KOGs through the phyletic range of eukaryotes strongly correlates with their functions and with the effect of gene knockout on the organism's viability. The approximately 40% of KOGs that are represented in six or seven species are enriched in proteins responsible for housekeeping functions, particularly translation and RNA processing. These conserved KOGs are often essential for survival and might approximate the minimal set of essential eukaryotic genes. The 131 single-member, pan-eukaryotic KOGs we identified were examined in detail. For around 20 that remained uncharacterized, functions were predicted by in-depth sequence analysis and examination of genomic context. Nearly all these proteins are subunits of known or predicted multiprotein complexes, in agreement with the balance hypothesis of evolution of gene copy number. Other KOGs show a variety of phyletic patterns, which points to major contributions of lineage-specific gene loss and the 'invention' of genes new to eukaryotic evolution. Examination of the sets of KOGs lost in individual lineages reveals co-elimination of functionally connected genes. Parsimonious scenarios of eukaryotic genome evolution and gene sets for ancestral eukaryotic forms were reconstructed. The gene set of the last common ancestor of the crown group consists of 3,413 KOGs and largely includes proteins involved in genome replication and expression, and central metabolism. Only 44% of the KOGs, mostly from the reconstructed gene set of the last common ancestor of the crown group, have detectable homologs in prokaryotes; the remainder apparently evolved via duplication with divergence and invention of new genes. CONCLUSIONS: The KOG analysis reveals a conserved core of largely essential eukaryotic genes as well as major diversification and innovation associated with evolution of eukaryotic genomes. The results provide quantitative support for major trends of eukaryotic evolution noticed previously at the qualitative level and a basis for detailed reconstruction of evolution of eukaryotic genomes and biology of ancestral forms. PMID- 14759259 TI - In silico identification and experimental validation of PmrAB targets in Salmonella typhimurium by regulatory motif detection. AB - BACKGROUND: The PmrAB (BasSR) two-component regulatory system is required for Salmonella typhimurium virulence. PmrAB-controlled modifications of the lipopolysaccharide (LPS) layer confer resistance to cationic antibiotic polypeptides, which may allow bacteria to survive within macrophages. The PmrAB system also confers resistance to Fe3+-mediated killing. New targets of the system have recently been discovered that seem not to have a role in the well described functions of PmrAB, suggesting that the PmrAB-dependent regulon might contain additional, unidentified targets. RESULTS: We performed an in silico analysis of possible targets of the PmrAB system. Using a motif model of the PmrA binding site in DNA, genome-wide screening was carried out to detect PmrAB target genes. To increase confidence in the predictions, all putative targets were subjected to a cross-species comparison (phylogenetic footprinting) using a Gibbs sampling-based motif-detection procedure. As well as the known targets, we detected additional targets with unknown functions. Four of these were experimentally validated (yibD, aroQ, mig-13 and sseJ). Site-directed mutagenesis of the PmrA-binding site (PmrA box) in yibD revealed specific sequence requirements. CONCLUSIONS: We demonstrated the efficiency of our procedure by recovering most of the known PmrAB-dependent targets and by identifying unknown targets that we were able to validate experimentally. We also pinpointed directions for further research that could help elucidate the S. typhimurium virulence pathway. PMID- 14759260 TI - Genome-wide investigation of light and carbon signaling interactions in Arabidopsis. AB - BACKGROUND: Light and carbon are two essential signals influencing plant growth and development. Little is known about how carbon and light signaling pathways intersect or influence one another to affect gene expression. RESULTS: Microarrays are used to investigate carbon and light signaling interactions at a genome-wide level in Arabidopsis thaliana. A classification system, 'InterAct Class', is used to classify genes on the basis of their expression profiles. InterAct classes and the genes within them are placed into theoretical models describing interactions between carbon and light signaling. Within InterAct classes there are genes regulated by carbon (201 genes), light (77 genes) or through carbon and light interactions (1,247 genes). We determined whether genes involved in specific biological processes are over-represented in the population of genes regulated by carbon and/or light signaling. Of 29 primary functional categories identified by the Munich Information Center for Protein Sequences, five show over-representation of genes regulated by carbon and/or light. Metabolism has the highest representation of genes regulated by carbon and light interactions and includes the secondary functional categories of carbon containing-compound/carbohydrate metabolism, amino-acid metabolism, lipid metabolism, fatty-acid metabolism and isoprenoid metabolism. Genes that share a similar InterAct class expression profile and are involved in the same biological process are used to identify putative cis elements possibly involved in responses to both carbon and light signals. CONCLUSIONS: The work presented here represents a method to organize and classify microarray datasets, enabling one to investigate signaling interactions and to identify putative cis elements in silico through the analysis of genes that share a similar expression profile and biological function. PMID- 14759261 TI - Predicting specificity in bZIP coiled-coil protein interactions. AB - We present a method for predicting protein-protein interactions mediated by the coiled-coil motif. When tested on interactions between nearly all human and yeast bZIP proteins, our method identifies 70% of strong interactions while maintaining that 92% of predictions are correct. Furthermore, cross-validation testing shows that including the bZIP experimental data significantly improves performance. Our method can be used to predict bZIP interactions in other genomes and is a promising approach for predicting coiled-coil interactions more generally. PMID- 14759262 TI - Versatile and open software for comparing large genomes. AB - The newest version of MUMmer easily handles comparisons of large eukaryotic genomes at varying evolutionary distances, as demonstrated by applications to multiple genomes. Two new graphical viewing tools provide alternative ways to analyze genome alignments. The new system is the first version of MUMmer to be released as open-source software. This allows other developers to contribute to the code base and freely redistribute the code. The MUMmer sources are available at http://www.tigr.org/software/mummer. PMID- 14759280 TI - Comparison of the psychological evaluation test and classical psychoanalysis in infertile women. AB - This study aims to compare a psychological evaluation test to classical psychoanalysis in infertile women. Two hundred women were submitted to the Psychological Evaluation Test (PET). The sum of the scores for the responses ranged from 15 to 60 points, with scores >/=30 points being defined as 'psycho emotional maladjustment' (cut-off point: median + 25%). For comparison, the patients were simultaneously submitted to a psychological examination by a psychologist, who was unaware of the PET results. Of the 200 patients, 66 (33%) presented a test with >/=30 points ('psycho-emotional maladjustment') and 134 (67%) a test with <30 points (normal). Upon psychological examination, 105 (52.5%) presented an abnormal evaluation and 95 (47.5%) a normal evaluation. For the PET, statistical analysis showed 82% efficiency, 62% sensitivity, 98% positive predictive value, 99% specificity, 70% negative predictive value, likelihood ratio for a positive test result 62, and likelihood ratio for negative test result 0.38. The PET proved to be a useful clinical instrument, being of help in the selection of patients with psychological needs induced by infertility. PMID- 14759281 TI - Mitochondria in reproduction. PMID- 14759282 TI - New approaches to the treatment of mitochondrial disorders. AB - Mitochondrial disorders are among the most common inherited metabolic diseases and the issue of treatment arises on a regular basis. There is no established treatment for mitochondrial disorders and current management is largely supportive, but recent advances in our understanding of the pathophysiology provide hope for novel treatments. Patients with mitochondrial myopathy due to mutations of mitochondrial DNA (mtDNA) may benefit from treatments that move normal mitochondrial genomes from the muscle satellite cells into skeletal muscle, but there are concerns about the long-term effects of this approach. A greater understanding of the pathophysiology of a number of nuclear genetic mitochondrial disorders suggests new avenues for treatment (such as copper histidine in children with SCO2 gene mutations, and strategies modifying intra mitochondrial nucleoside pools in the various disorders of mtDNA maintenance). A number of different strategies are also being explored at the molecular level, including the use of antigenomic molecules to mutated mtDNA and the allotropic expression of mutated mtDNA genes within the cell nucleus. Nuclear transfer techniques also provide hope for women at risk of transmitting pathogenic mtDNA mutations. PMID- 14759285 TI - The potential risks of abnormal transmission of mtDNA through assisted reproductive technologies. AB - The recent introduction of more invasive assisted reproductive techniques offers the possibility to provide a wider treatment profile to patients. However, some of these technologies are of considerable concern as they are fraught with the possible transmission of genetic abnormalities to the offspring they create. To date, much analysis of these technologies has been conducted at the chromosomal DNA level. While some analysis has been conducted on the extranuclear, mitochondrial genome (mtDNA), this has been mainly descriptive. In the vast majority of cases, it appears that mtDNA is maternally inherited. The impact that leakage of sperm mtDNA transmission might have for the offspring is discussed in the light of the recent identification of sperm mtDNA presence in a patient with mtDNA disease. The implications of introducing donor mtDNA into a recipient oocyte through both cytoplasmic and nuclear transfer are also discussed. Again, the implications for offspring survival are discussed and suggestions made as to why the techniques might provide valuable insights into mtDNA transmission, replication and transcription. In order to be confident that patients and their offspring are being offered safe treatment, it is argued that potentially some of these treatments may be of considerable benefit in the future but significant scientific research is required before these treatments can be effectively employed in the clinic. PMID- 14759284 TI - Degradation of paternal mitochondria after fertilization: implications for heteroplasmy, assisted reproductive technologies and mtDNA inheritance. AB - Maternal inheritance of mitochondrial DNA has long been regarded as a major paradox in developmental biology. While some confusion may still persist in popular science, research data clearly document that the paternal sperm-borne mitochondria of most mammalian species enter the ooplasm at fertilization and are specifically targeted for degradation by the resident ubiquitin system. Ubiquitin is a proteolytic chaperone that forms covalently linked polyubiquitin chains on the targeted proteinaceous substrates. The polyubiquitin tag redirects the substrate proteins to a 26-S proteasome, a multi-subunit proteolytic organelle. Thus, specific proteasomal inhibitors reversibly block sperm mitochondrial degradation in ooplasm. Lysosomal degradation and the activity of membrane lipoperoxidating enzyme 15-lipoxygenase (15-LOX) may also contribute to sperm mitochondrial degradation in the ooplasm, but probably is not crucial. Prohibitin, the major protein of the inner mitochondrial membrane, appears to be ubiquitinated in the sperm mitochondria. Occasional occurrence of paternal inheritance of mtDNA has been suggested in mammals including humans. While most such evidence has been widely disputed, it warrants further examination. Of particular concern is the documented heteroplasmy, i.e. mixed mtDNA inheritance after ooplasmic transplantation. Intracytoplasmic sperm injection (ICSI) has inherent potential for delaying the degradation of sperm mitochondria. However, paternal mtDNA inheritance after ICSI has not been documented so far. PMID- 14759287 TI - Spindles, mitochondria and redox potential in ageing oocytes. AB - Studies of human oocytes obtained from women of advanced reproductive age revealed that spindles are frequently aberrant, with chromosomes sometimes failing to align properly at the equator during meiosis I and II. Chromosomal analyses of donated and spare human oocytes and cytogenetic and molecular studies on the origin of trisomies collectively suggest that errors in chromosome segregation during oogenesis increase with advancing maternal age and as the menopause approaches. Disturbances in the fidelity of chromosome segregation, especially at anaphase I, leading to aneuploidy are prime causes of reduced developmental competence of embryos in assisted reproduction, as well as being responsible for the genesis of genetic disease. This review provides an overview of spindle formation and chromosome behaviour in mammalian oocytes. Evidence of a link between abnormal mitochondrial function in oocytes and somatic follicular cells, and finally disturbances in chromosome cohesion and segregation, and cell cycle control in aged mammalian oocytes, are also discussed. PMID- 14759289 TI - Unsuccessful oocyte retrieval: technical artefact or genuine 'empty follicle syndrome'? AB - Unsuccessful oocyte retrieval after apparently successful ovarian stimulation (also referred to as 'empty follicle syndrome') occurs in 1-7% of women undergoing assisted reproductive techniques. A literature review was performed, as individual studies have reached differing conclusions on the aetiology and treatment or management of the phenomenon. The aetiology is not clear, but probably multifactorial, and occurs in natural and stimulated cycles. In many cases, technical problems such as errors in human chorionic gonadotrophin (HCG) administration or defects in HCG batches can be identified, but this is not sufficient to account for all reported cases. The term empty follicle syndrome is inappropriate in cases in which such procedural factors can be identified. In many patients, however, unsuccessful oocyte retrieval appears to be due to an underlying ovarian dysfunction, and some may have a genuine empty follicle syndrome. Appropriate measures, such as monitoring of serum beta-HCG, should be taken to minimize the risk of unsuccessful oocyte retrieval. This review discusses the potential causes of unsuccessful oocyte retrieval, its clinical implications, and potential solutions to this clinical problem. PMID- 14759290 TI - Effect of phenobarbital on the induction of glutathione S-transferases in rat testis. AB - Glutathione S-transferases are a family of multifunctional proteins involved in intracellular transport processes and drug detoxication. In rats, these enzymes are dimeric proteins, and exist as cytosolic and microsomal proteins. The affinity purified rat testicular glutathione S-transferases are comprised of four subunits, Yc of alpha class, Yb and Ybeta of mu class and Ydelta of pi class. On chromatofocusing, they were resolved into six anionic and four cationic isozymes. The cationic isozymes were found to be abundant. All these isozymes on electrophoresis were found to contain heteromers except for two isozymes. The expression of individual subunits and their activity was elevated on treatment with multiple doses of phenobarbital in rat testis. Among all of these, according to the immunological studies, Ydelta, a pi class glutathione S-transferase, was induced predominantly in response to phenobarbital. PMID- 14759291 TI - Cytoplasmic transfer in the mouse in conjunction with intracytoplasmic sperm injection. AB - Cytoplasmic transfer (CT) from a donor to a recipient oocyte has been used clinically to facilitate human pregnancies. Data reported here describe the first characterization of CT coincident with intracytoplasmic sperm injection in the mouse system. Sibling oocytes were used to transfer 2, 4, or 6 pl of ooplasm to a recipient egg along with a sperm head using piezo-actuated injection. Survival and fertilization after CT were comparable to controls at 2 pl and 4 pl, but survival was significantly reduced with 6 pl volumes. Development to the blastocyst stage was also inversely related to CT volume, with some decline beginning with the 4 pl CT group. However, some blastocysts did develop in all of the groups. The results are in contrast with human eggs, which tolerate larger CT volumes. Results indicate that the mouse system can be used to characterize the transfer of exogenous materials concomitant with sperm injection, provided that the CT volume is not excessive. PMID- 14759293 TI - Differences in chromosome susceptibility to aneuploidy and survival to first trimester. AB - The purpose of this study was to find specific rates of aneuploidy in cleavage stage embryos compared with first trimester data and to evaluate post-zygotic selection against aneuploidy. A total of 2058 embryos were analysed by flurorescence in-situ hybridization (FISH), and specific aneuploidy rates were obtained for 14 chromosomes. Data from morphologically abnormal embryos could be pooled with data from preimplantation genetic diagnosis (PGD) cycles because it was observed that they had similar rates of aneuploidy; thus, for the purpose of studying aneuploidy they could be, and were, pooled. Specific chromosome aneuploidy rates were not related to morphology or development of the embryos. The average maternal age of patients with aneuploid embryos was significantly higher than the overall analysed population. Monosomy appeared more commonly than trisomy. The chromosomes most frequently involved in aneuploidy were (in order) 22, 16, 21 and 15. When compared with first trimester pregnancy data, aneuploidies detected at cleavage stage seem to die in excess of 90% before reaching first trimester, with the exception of chromosome 16 and gonosomes (76% and 14% respectively). Differences in chromosome-specific aneuploidy rates at first trimester conceptions are probably produced by different chromosome specific aneuploidy rates at cleavage stage and different survival rates to first trimester. PMID- 14759294 TI - Placental protein 14 concentrations in circulation related to hormonal parameters and reproductive outcome in women undergoing IVF/ICSI. AB - Serum concentrations of placental protein 14 (PP14), steroids and gonadotrophins were related to the outcome of IVF/intracytoplasmic sperm injection in 195 normogonadotrophic women subjected to the long protocol gonadotrophin-releasing hormone agonist (GnRHa; buserelin) pituitary down-regulation protocol and gonadotrophin stimulation (HMG or rFSH). Pituitary down-regulation was initiated on cycle day 21 and the patients were randomized to either intranasal or s.c. administration of buserelin. After 14 days of down-regulation, the patients were randomized on stimulation day 1 (S1) to ovarian stimulation with 225 IU per day of either human menopausal gonadotrophin (HMG) or recombinant FSH (rFSH) for a fixed period of 7 days. The daily gonadotrophin dose was adjusted on the following day according to ovarian response. Patient's blood was sampled for PP14 and hormone analysis on cycle days 21, S1, S8 and on the day of oocyte retrieval. Mean concentrations of PP14 on day 21 of the cycle were significantly lower in conception than in non-conception cycles, whereas progesterone and oestradiol were similar in conception and non-conception cycles. PP14 concentrations on the first day of stimulation and at oocyte retrieval were significantly higher in conception than in non-conception cycles, whereas concentrations after 8 days of stimulation were similar. Neither mode of GnRHa administration nor type of gonadotrophin significantly influenced PP14 concentrations throughout ovarian stimulation. Circulating PP14 is thus an important physiological signal of the fertility status of the individual in the cycle antecedent to and during ovarian stimulation. Measuring mid-luteal serum PP14 may offer a clinical test helping to decide if infertility treatment should be initiated in the subsequent cycle. PMID- 14759296 TI - Averting abnormal inheritance: potential of gene therapy and preimplantation diagnosis. AB - Serious inherited disease in children can be averted by preimplantation genetic diagnosis (PGD) and potentially by gene therapy in addition to prenatal diagnosis. PGD is now well established and provides a secure, if expensive and complex form of care. Gene therapy has been practised only in animals, although its success in alleviating various conditions in adults and newborns, together with the scientific drive of the genome project, make it a highly likely approach over coming years. Pros and cons of both approaches are contrasted and compared. Newer reproductive techniques such as somatic cell hybridization promise to add new dimensions to gene therapy, and could be combined with PGD. This paper discusses the finer details of these options, their safety and the ethical issues they have raised. PMID- 14759297 TI - Towards preimplantation diagnosis of cystic fibrosis using microarrays. AB - Cystic fibrosis (CF) is a common indication for preimplantation genetic diagnosis (PGD). A 3-bp deletion (DeltaF508) in the cftr gene, which accounts for approximately 80% of all CF mutations in the Caucasian population, is normally diagnosed in IVF embryos using fluorescent PCR (FL-PCR) and allelic sizing. In PGD, the possibility of using microarrays for genetic diagnosis is largely unexplored. Therefore, the aim of this study was to prove the diagnostic capability of microarrays for PGD, using DeltaF508 as a model mutation. To this end, oligonucleotide probes representing both the normal and DeltaF508 disease alleles were used to construct a single microarray platform. Target DNA, which was generated by PCR and labelled with the fluorescent dye Cy3, was hybridized to the array and the DeltaF508 genotypes assigned from the fluorescence bound to each allelic probe. The performance of the array was evaluated by its ability to detect DeltaF508 mutations in target DNA. Strong binding of the target to the probes was observed, allowing the expected DeltaF508 genotypes to be assigned. The reliability and accuracy of the microarray diagnosis for DeltaF508 was blindly assessed on 10 samples with either a homozygous normal, homozygous affected or heterozygous genotype. All samples were correctly genotyped. In addition, PCR products from a previous PGD case involving DeltaF508 were re evaluated on the array, with results in complete concordance with allelic sizing methods used to make the original diagnosis. Together, these findings prove the concept that the DeltaF508 mutation of CF can be reliably and accurately diagnosed at the single cell level using microarray analysis. The availability of more cost-effective array platforms comprising mutation probes for common single gene disorders and a reliable method of whole genome amplification (WGA) would allow PGD to be offered to the majority of PGD patients with minimal or no change to methodology. PMID- 14759299 TI - Endometrial thickness cannot predict ongoing pregnancy achievement in cycles stimulated with clomiphene citrate for intrauterine insemination. AB - To date, limited data exist concerning the relation between endometrial thickness on the day of human chorionic gonadotrohin (HCG) administration and ongoing pregnancy achievement in cycles stimulated with clomiphene citrate for intrauterine insemination (IUI). In a prospective study, 168 couples were stimulated with clomiphene citrate from day 3 to day 7 of the cycle and endometrial thickness was assessed by ultrasound three times on the day of ovulation triggering. Ovulation was induced with HCG as soon as >/=1 follicle of >/=17 mm was present at ultrasound independently of endometrial thickness. IUI was performed 36 h after HCG administration. The main outcome measure was ongoing pregnancy. No difference was observed in endometrial thickness between patients who did or did not achieve an ongoing pregnancy (7.6 +/- 0.3 versus 7.6 +/- 0.2 respectively; P = 0.7). No discriminative ability of endometrial thickness on the achievement of ongoing pregnancy could be shown by receiver operating characteristic (ROC) curve analysis (area under the ROC curve 0.51, 95% CI: 0.44 0.59). In conclusion, endometrial thickness cannot predict ongoing pregnancy achievement in IUI cycles stimulated with clomiphene citrate. PMID- 14759300 TI - Different preferences for prenatal diagnosis in pregnancies following assisted reproduction versus spontaneous conception. AB - The uptake of prenatal diagnosis in 436 singleton and 146 twin pregnancies following assisted reproduction was compared with a matched group of spontaneous conceptions. The first and second trimester ultrasound examination included target fetal anatomic evaluation and screening by specific markers described for fetal aneuploidy. Women with assisted conception attended significantly more often for first trimester prenatal diagnosis (57.9 versus 34.9%, P < 0.01), but had fewer examinations in the early second trimester at 15-18 weeks (37.8 versus 48.8%, P < 0.01) than those with spontaneous conception. Screen positive results of 6.5 and 6.9% for first trimester examination and 6.0 and 7.3% for second trimester examination were found in assisted conceptions and controls respectively. A significantly higher rate of invasive prenatal diagnosis was observed in the second trimester for spontaneous conceptions, 20.0 versus 11.8% (P < 0.01) compared with assisted conceptions. This was attributed to the higher rate of invasive procedures in advanced maternal age >/=35 years of 40.7 versus 28.6% (P = 0.01) in spontaneous and assisted conceptions respectively. With the purpose of avoiding invasive testing, women with assisted conception were more likely to use the results of the ultrasound examination to guide their final decision about invasive testing, rather than undergo genetic amniocentesis as a first option. PMID- 14759302 TI - Multiple gestation as a marker of reproductive efficacy: learning from assisted reproductive technologies. AB - This study postulates that apart from the number of embryos transferred, women with multiple gestation represent a subgroup of highly fertile individuals, whose embryos implant with higher efficiency than women with single gestation. Furthermore, each embryo generated from these women has a higher chance of reaching full term. The objective of this study was to compare implantation rate with the outcome of pregnancy (up to week 20) in multiple gestations following assisted reproductive techniques. The study group comprised 162 women with multiple gestation after assisted reproduction, followed prospectively with at least three ultrasound examinations performed between weeks 5 and 20 after the last menstrual period. Control group A comprised 344 fertile women with spontaneous single pregnancy followed with transvaginal ultrasound. Control group B consisted of 317 infertile women conceiving with single gestation after assisted reproduction and followed prospectively as in the study group. Embryo implantation rate and spontaneous embryo/fetal reduction, either partial or total (abortion), were registered in each case. Overall implantation in women with multiple gestation was higher (54.6%) than in the corresponding controls (25.6%). Furthermore, spontaneous embryo/fetus reduction was similar in the study cases and in fertile women (12.6 and 10.8% respectively) and significantly smaller than in the control group B (20.8%). Women with high reproductive efficacy exposed to assisted reproductive techniques generate cohorts of good quality embryos, with a high chance of implantation and of reaching birth. PMID- 14759303 TI - [Strengthen the prevention and treatment of complications to improve the survival rate and quality of life in preterm infants]. PMID- 14759304 TI - [Treatment of influenza in children]. PMID- 14759305 TI - [The enteral feeds in very low birthweight infants]. AB - OBJECTIVE: The study aimed to investigate the clinical characteristics of enteral feeding in very low birthweight infants (VLBWI), to determine the risk factors associated with feeding intolerance, and to analysis the beneficial factors in order to improve gut motility and maturation. METHODS: The study was carried out in 38 VLBWI, birthweight (1,314 +/- 180) g, in the NICU of authors' department. They were divided into feeding tolerance and intolerance groups, and earlier enteral feeding and later groups. Comparison was made between two groups about the associate factors. RESULTS: The incidence of feeding intolerance was 55 per cent. There was a significant difference in two groups about the clinical factors (gestational age, birth weight, the age of the first feeding, time of full enteral feeding and the hospitalized days). The significant risk factors associated with feeding intolerance were the smaller gestational age, umbilical catheterization, theophylline therapy, and delay of the time when the first feed was commenced. CONCLUSION: If vital signs are stable, trophic feeding in VLBWI should be commenced as soon as possible during the first 6 days of life. Early trophic feeding, slowly increase the feeding volume, carefully fast, and moving bowel were suggested and will improve feeding tolerance and gastrointestinal maturation. PMID- 14759306 TI - [Effects of intermittent nasogastric feeding with nonnutritive sucking on nutrient and gastrointestinal tract transit time in premature infants]. AB - OBJECTIVE: To evaluate the effects of nonnutritive sucking (NNS) on the nutrient intake, physical growth, feeding-related complications and whole gastrointestinal transit time (WGTT) in premature infants. METHODS: Thirty eight healthy appropriate for gestational age premature infants (birth weights ranged from 1 050 g to 1 790 g) accepting intermittent nasogastric feeding (INGF) were randomized into NNS group and N-NNS group according to INGF with and without NNS and fed with the same milk formula. The following data were collected and recorded, the physical growth parameters (e.g, body weight, length and head circumference) and the birth-weight regaining time, the fluid intake (including both intravenous and oral), caloric intake, time of reaching 418.4 kJ/(kg.d) by enteral feeding, time of putting nasogastric tube, stool frequency and characters, and relevant complications. WGTT were monitored. RESULTS: The birth weight regaining time in NNS group was significantly shorter than that in N-NNS group [(8.8 +/- 3.7) d vs (11.1 +/- 3.0) d, P < 0.05]. Within two weeks after feeding, there was no significant difference in the increase of body weight, length and head circumference between the two groups (P > 0.05). The time of reaching 418.4 kJ/(kg.d) by enteral feeding in NNS group was significantly shorter than that in N-NNS group [(12.3 +/- 5.1) d vs (15.7 +/- 5.2) d, P < 0.05]; the times of putting nasogastric tube were respectively (13 +/- 10) d and (17 +/- 12) d, but the difference was not significant (P > 0.05). The morbidity of such complications as vomiting and abdominal distension was lower in NNS group than that in N-NNS group, but the difference was not statistically significant (P > 0.05). However, the morbidity of gastric residue in NNS was significantly lower than that in N-NNS (P < 0.05). WGTT of the second week in NNS group was significantly shorter than that in N-NNS [(33 +/- 13) h vs (45 +/- 20) h, P < 0.05]. Stool frequencies of the second week in NNS group were significantly more than those in N-NNS group [(2.26 +/- 0.17) times/d vs (1.79 +/- 0.58) times/d, P < 0.05]. However, there were no significantly differences in WGTT and stool frequencies of the first week between the two groups (P > 0.05). CONCLUSION: NNS was recommended as a beneficial intervention for premature infants during intermittent nasogastric tube feeding. PMID- 14759307 TI - [Lung protective strategies of ventilation in respiratory distress syndrome of neonates]. AB - OBJECTIVE: The authors compared the protective ventilation strategies with the assistance of pulmonary mechanic measurements (PM) to the conventional ventilation practice in order to investigate the incidence of VALI, explore the best ventilator settings in RDS and the clinical effects of hypercapnia in neonates. METHODS: One hundred and ten newborn infants, diagnosed as respiratory distress syndrome of neonatal and needed ventilator support, were recruited from the intensive care unit from 1994 to 2001. Infants with RDS were divided into non pulmonary mechanics measurement (NPM) group and pulmonary mechanics measurement (PM) group. The ventilator settings, artery blood gas analysis, ventilator associated lung injury and other clinical data were analyzed and compared. RESULTS: In the PM group, expiratory tidal volume of (7.70 +/- 1.05) ml/kg, C(20)/C of (1.98 +/- 0.92) and time constant of (0.09 +/- 0.03) sec were measured. Ventilator setting in PM group [PIP of (26.7 +/- 1.7) cmH(2)O, MAP of (11.9 +/- 2.0) cmH(2)O, Ti of (0.45 +/- 0.10) sec] were significantly less than those in NPM group [PIP of (30.5 +/- 3.4) cmH(2)O, MAP of (14.9 +/- 3.4) cmH(2)O, Ti of (0.75 +/- 0.10) sec P < 0.001, respectively]. No differences were noted in demographic data between the two groups regarding to the pH (7.30 +/- 0.04 vs 7.31 +/- 0.10), PaO(2) [(59 +/- 16) vs (57 +/- 17) mmHg], heart rate [(145 +/- 6) vs (144 +/- 8) bpm] and blood pressure [(39.0 +/- 3.6) vs (40.0 +/- 4.6) mmHg], except for PaCO(2) in PM group which was significantly higher than that in NPM group [(48 +/- 6) vs (40 +/- 10) mmHg, P < 0.001]. OI in PM group was significantly less than that in NPM group (14 +/- 8 vs 19 +/- 13, P < 0.05). Incidence of ventilator-associated lung injury in PM group was significantly lower than that in NPM group (13% vs 32%, P < 0.05). No differences were noted in demographic data between the two groups regarding to the length of ventilator support [(4.2 +/- 1.7) vs (3.9 +/- 1.8) day], length of supplemental oxygen requirements [(13 +/- 7) vs (11 +/- 7) day], length of hospitalization [(22 +/- 11) vs (19 +/- 14) day], mortality (8% vs 14%), incidence of PDA (33 vs 36%) and IVH (40% vs 42%). CONCLUSIONS: The assistance use of pulmonary mechanics measurement can guide the ventilator adjustment and decrease the ventilator associated lung injury in neonatal RDS. The low PIP, low tidal volume, shorter inspiratory time and adequate PEEP strategies in the treatment of RDS showed no significant effects on the oxygenation in neonates. The mild permissive hypercarbia in newborn did not increase the incidence of IVH. PMID- 14759308 TI - [Effects of recombinant human erythropoietin on the immune function of premature rats]. AB - OBJECTIVE: To observe the effect of recombinant human erythropoietin (rHuEPO) on immune function of premature rats. METHODS: RHuEPO of 250 IU/(kg.t) or 500 IU/(kg.t) was administered to premature rats every other day for nineteen days. The control premature rats were received normal saline. The changes of hemoglobin (Hb), serum erythropoietin (EPO), red blood cell (RBC) immune function, T lymphocyte proliferative responsiveness, and production of tumor necrosis factor alpha (TNF-alpha) were observed. RESULTS: Premature rats showed lower levels on Hb, RBC immune function, T cell responsiveness and production of TNF-alpha compared with mature rats at birth. The postnatal declines of Hb and RBC immune function were lessened in the treated groups of premature rats, the higher dosage group of 500 IU/(kg.t) was more significant than the lower dosage group of 250 IU/(kg.t). When experiments were over, Hb of control premature rats was (7.72 +/- 0.89) g/dl, Hb of premature rats received 500 IU/(kg.t) was (10.08 +/- 0.90) g/dl (P < 0.01). C3b-R% of control premature rats was (11.00 +/- 0.95)%, C3b-R% of premature rats received 500 IU/(kg.t) was (17.75 +/- 1.04)% (P < 0.01). IC-R% in control premature rats was (12.83 +/- 1.33)%, IC-R% of premature rats received 500 IU/(kg.t) was (10.50 +/- 1.67)% (P < 0.01). The postnatal rise of T cell responsiveness and the production of TNF-alpha in premature rats increased in the treated groups, which was more significant in the higher dosage group of 500 IU/(kg.t) than in the lower dosage group of 250 IU/(kg.t). The OD index of control premature rats was 0.159 +/- 0.014, the OD index of premature rats received 500 IU/(kg.t) was 0.354 +/- 0.050 (P < 0.01). TNF-alpha in control premature rats was (0.270 +/- 0.014) ng/ml, TNF-alpha of premature rats received 500 IU/(kg.t) was (0.415 +/- 0.010) ng/ml (P < 0.01). CONCLUSIONS: (1) Premature rats had lower RBC immune function and T cell responsiveness and underproduction of TNF-alpha at birth. (2) Premature rats had an improvement with the RBC immune function after rHuEPO administration. (3) Premature rats had improvements with T cell responsiveness and TNF-alpha after rHuEPO administration, and there was a positive correction between the RBC immune function and T cell responsiveness with the production of TNF-alpha. PMID- 14759309 TI - [Changes of tissue factor and tissue factor pathway inhibitor in neonatal jaundice due to infection]. AB - OBJECTIVE: Tissue factor (TF) is an important factor in extrinsic coagulation. Tissue factor pathway inhibitor (TFPI) is a negative regulator of coagulation mediated by TF. Studies on TF and TFPI focus mainly on adult objects, seldom have been done on newborns, especially on sick newborns. The aim of this study was to observe the changes of TF and TFPI in plasma of newborns with infection jaundice and to research the effect of jaundice and infection on the balance of TF and TFPI in newborns. METHODS: The content of TF and TFPI in plasma of 21 jaundiced newborns with infection and 8 jaundiced newborns without infection as control was determined quantitatively with the enzyme-linked immunosorbent assay (ELISA). RESULTS: The content of TFPI and TF in plasma of jaundiced newborn with infection was significantly higher than that of controls [TFPI (21.0 +/- 4.3) vs. (16.2 +/- 1.9) microg/L, P < 0.01; TF (177 +/- 79) vs. (51 +/- 24) ng/L, P < 0.01]. The ratio of TFPI/TF was significantly lower in newborn with infection jaundice than the controls (137 +/- 61 vs. 319 +/- 67, P < 0.01). The 21 jaundiced newborns with infection were divided into the severe hyperbilirubinemia group (serum bilirubin > or = 205.2 micromol/L, n = 10) and the mild hyperbilirubinemia group (serum bilirubin < 205.2 micromol/L, n = 11). There was no significant difference of TFPI level between the severe hyperbilirubinemia group and mild hyperbilirubinemia group (P > 0.05). The TF content in the severe hyperbilirubinemia group was higher than that in the mild hyperbilirubinemia group (216 +/- 79 vs.141 +/- 63, P < 0.01), while the ration of TFPI/TF was lower in the severe hyperbilirubinemia group than in the mild hyperbilirubinemia group (100 +/- 30 vs. 171 +/- 74, P < 0.01). CONCLUSION: Infection might induce imbalance between the coagulation inhibition and activation in newborns. Hyperbilirubinemia can aggravate the imbalance induced by the infection through increasing plasma TF level. PMID- 14759310 TI - [Cholesterol concentrations in cord blood of newborn infants]. AB - OBJECTIVE: Familial hypercholesterolemia (FH) is an autosomal dominant disease with an estimated worldwide prevalence of 0.2%. It is caused by a multitude of low density lipoprotein receptor gene mutations. It is characterized with high levels of total cholesterol (TC), low density lipoprotein cholesterol (LDL-C) and a high incidence of coronary artery disease in young adults. Cord blood cholesterol concentration is used for mass screening of FH. The purpose of this study was to detect the lipid levels of cord blood in newborn infants from China and to determine the cut-off point after 1 to 2 years follow-up. METHODS: TC, triglycerides (TG), LDL-C and high density lipoprotein cholesterol (HDL-C) were determined in 242 healthy full-term newborn infants. RESULTS: The mean values of TC, TG, LDL-C and HDL-C in cord blood were (1.69 +/- 0.40) mmol/L, (0.23 +/- 0.12) mmol/L, (0.81 +/- 0.21) mmol/L and (0.58 +/- 0.16) mmol/L (mean +/- standard deviation), respectively. The HDL-C concentration in male neonates was lower than that in female neonates (P < 0.05). CONCLUSION: After the follow-up of 1 to 2 years for FH, the recommended screening cut-off points were TC > or = 2.47 mmol/L and LDL-C > or = 0.89 mmol/L. PMID- 14759311 TI - [Early diagnosis, prophylaxis and treatment of intraventricular hemorrhage in premature infants]. PMID- 14759312 TI - [Summary of the 3rd Bi-coastal symposium on neonatology and the 6th national symposium of neonatology]. PMID- 14759313 TI - [Summary of 2002 national conferences of pediatric cardiac arrhythmia and cardiovascular diseases]. PMID- 14759314 TI - [Summary of the 11th national conference of pediatric nephrology]. PMID- 14759315 TI - [A case report of lead encephalopathy]. PMID- 14759316 TI - [Treatment of Wilson's disease with penicillamine and zinc salts: a follow-up study]. AB - OBJECTIVE: Wilson's disease (WD) is an autosomal recessive disorder characterized by excessive accumulation of copper in the liver and later in the brain and other organs. Penicillamine acts as a reductive chelator. Zinc salts induce the synthesis of metallothionein in cells. Thus these two drugs are theoretically synergistic for the treatment of the disease. However, the two drugs may also have some unfavorable interactions. In this study, the effect of the therapy with combined penicillamine and zinc salts was evaluated based on the follow-up observations of 21 patients with Wilson's disease. METHODS: Using the combined therapy of penicillamine [10-30 mg/(kg.d)] and zinc (22.5 mg, 3 times per day), follow-up study by hospitalization or communication with telephone or mail. RESULTS: Before treatment, all the 21 patients were suffered from chronic liver disorder. Among them, 13 patients (62%) showed to be reactive to the treatment for their liver disorder, 5 patients (24%) died, and 3 patients (14%) dropped off our follow-up study. Among the 5 patients who died, 3 died within 40 days after treatment, one had taken penicillamine only 8 mg/(kg.d), and one died after discontinuation of the treatment by the parents. Of the 12 patients having neurological involvement, neurological symptoms disappeared or markedly improved in 11 patients after treatment. One patient dropped off the follow-up study. The patient with renal tubular acidosis responded well to the treatment. Urine routine analysis was followed up in 6 of the 7 patients with hematuria. Hematuria disappeared in one, became less severe in 1, and remained unchanged in 4 patients. Hypersensitivity to penicillamine was found in one patient. WBC and platelet were found decreased further in 3 patients after the medications. CONCLUSIONS: The combined therapy with penicillamine and zinc salts was effective in treatment of patients with Wilson's disease. PMID- 14759317 TI - [Bismuth subcitrate caused acute renal failure in a child]. PMID- 14759318 TI - [A nationwide survey in China on prevalence of asthma in urban children]. AB - OBJECTIVE: Asthma is one of the most common chronic disorders in childhood. The prevalence rate of asthma and other atopic diseases have increased steadily in many countries. The present survey was conducted to find out the prevalence of childhood asthma in urban areas in China, and to assess the influence of asthma on patients' social life and their families. METHODS: A nationwide randomized survey, covering 43 cities in 31 provinces, on the prevalence of childhood asthma was carried out by the National Pediatric Cooperative Group on Asthma Research from June to October 2000, including a population of 437,873 children aged 0-14 years. Patients who had asthma, infantile asthma, cough variant asthma and questionable asthma (including asthmatic bronchitis) were chosen as subjects. RESULTS: Totally 10,065 subjects were screened out as asthma associated children by means of questionnaire to the parents, physical examination and case history review. Among them 7401 (73.53%) children, aged 3 years or more, were diagnosed as asthma of children; 1109 (11.02%) infants and young children, aged less than 3 years, as asthma of infants and young children; 785 (7.80%) as cough variant asthma; and 770 (7.65%) as questionable asthma. The prevalence of the average accumulated asthma of all the 0-14 years old asthma population (including asthma of older children and of infants) in the 31 provinces, 43 cities was 1.97%. Male/female ratio was 1.75:1. There was statistically significant difference in asthma prevalence among the 43 cities, with the highest rate in Chongqing (4.63%) and the lowest in Xining (0.25%). Overall, within China, the prevalence rate in North China was relatively lower (0.99%) and was higher in South China (1.54%), the highest was seen in eastern areas of China (2.37%), 4670 patients (70.00%) of the children had their onset before three years of age. The current 2 years prevalence of Chinese urban children was 1.54%. Thirty-six of the 43 cities had received the same kind of asthma prevalence survey 10 years ago, which made a longitudinal comparison of the two surveys available. For 6370 patients (95.47%) the family members' work was affected because of their asthma attacks. One third of the patients had used inhaler corticosteroids. In about 2/3 of patients the diagnosis of asthma was correct, while only 1/3 patients' cough variant asthma was correct compared with their early diagnosis. Sixteen percent of patients were never diagnosed to have asthmatic diseases previously. CONCLUSIONS: There has been a significantly increasing trend of asthma prevalence during the ten years, especially in the older age group. From the data of present survey it was inferred that there has been certain improvement, although not ideal, in the accuracy of diagnosis and in the practice of steroid inhalation therapy by the pediatricians in different cities. PMID- 14759319 TI - [Genetic diversity of human Parvovirus B19 VP1 unique region]. AB - OBJECTIVE: Human Parvovirus B19 (HPV B19) is a small (23 nm), non-enveloped DNA virus found in 1974. It has been proved that HPV B19 is associated with a variety of childhood diseases, such as erythema infectious, transient aplastic crisis, aplastic anemia, idiopathic thrombocytopenic purpura and arthropathy, etc. There have been no any effective vaccines to prevent HPV B19 infection so far. The HPV B19 genome is composed of 5.6 kb single strand DNA. This genome encodes a nonstructural protein NS1, two structural proteins VP1 and VP2. Most neutralizing linear epitopes of HPV B19 cluster in the VP1 unique and VP1-VP2 junction regions. Only proteins encoded by genes of the VP1 unique and VP1-VP2 junction regions can stimulate bodies to produce protective antibodies. Aim of the present study was to get the VP1 unique region gene of HPV B19 and to analyze the genetic diversity so as to further study its function and application. METHODS: The VP1 unique region gene of HPV B19 was amplified from the serum of a child with idiopathic thrombocytopenic purpura by PCR. The purified PCR product was cloned into pGEM-T easy vector and transfected into the host strain E. coli (DH5 alpha). Positive clones were chosen and then the target gene was sequenced. RESULTS: The target gene sequence of HPV B19 VP1 unique region was amplified and cloned successfully. It had 705 nucleotides. Compared with the relevant sequences published in Genbank, the sequencing results were revealed with two nucleotides changes in the HPV B19 VP1 unique region, but their coding amino acid were not changed. CONCLUSION: It is suggested that genetic diversity exists in the VP1 unique region of HPV B19. Construction of the recombinant plasmid of HPV B19 VP1 unique region gene might benefit to further study. PMID- 14759320 TI - [Results of Wilms' tumor trial (WT-99) in Shanghai children's medical center]. AB - OBJECTIVE: Wilms' Tumor Trial (WT-99) of Shanghai Children's Medical Center was designed and conducted by applying therapeutic regimens stratified by stage and histology in accordance with National Wilms' Tumor Study (NWTS) criteria of U.S.A. The main aim of WT-99 was to reduce treatment of low-stage, favorable histology (FH) tumors without impairing survival and to improve prognosis of stage III and IV (FH) and unfavorable-histology (UFH) tremors with more intensive chemotherapy. METHODS: Diagnosis and treatment was decided by the multi disciplinary team including oncologists, surgeons, pathologists, radiologists and diagnostic radiologists. Twenty consecutively diagnosed patients were recruited between October 1998 and October 2002. The regimen for patients at favorable histology (FH) stage I and II and anaplastic stage I was vincristine (Vcr) and dactinomycin (Act-D) only, while for those at focal anaplastic stage II to IV and FH stage III and IV the regimen was Vcr, Act-D and adriamycin (Adr). Patients at diffuse anaplastic stage II to IV and clear cell stage I to IV received four-drug regimen including Vcr, etoposide (VP16), Adr and cytoxan (CTX). For those at rhabdoid stage I to IV the regimen was carboplatin, VP-16 and CTX. Un-resectable patients received 2 courses of Ifosfamide, Vcr and VP-16 as pre-surgery therapy. No radiation therapy was used for patients at stage I and FH stage II. RESULTS: Twenty patients, from 7 months to 12 years old, were enrolled. Pathologic analysis showed fourteen cases were at their FH, three at unfavorable-histology (UFH), two at clear cell and one at rhabdoid stage. Five patients were at stage I, five at stage II, six at stage III, three at stage IV and one at stage V. Eighteen reached complete response (90%), and two failed. One relapsed after 24 months of CCR and reached the second CR after intensive chemotherapy. No therapy related death happened. Survival rate (SR) was 90% (18/20) and event-free survival (EFS) was 85% (17/20) at 11-45 months, average 27 months. CONCLUSION: Multi-disciplinary team work model and protocol WT-99 are safe and effective for Wilms' tumor. PMID- 14759321 TI - [Efficacy and safety of one dose ceftriaxone vs. ten-day oral amoxicillin for treatment of acute otitis media in children]. AB - OBJECTIVE: To compare the efficacy and safety of a single ceftriaxone injection with 10-day oral amoxicillin in the treatment for children's acute otitis media. METHODS: This study was a prospective, comparative, open randomized, multicenter trial. In the ceftriaxone group, a single dose sodium ceftriaxone (50 mg/kg, total dose < 1 g) was injected. In the amoxicillin group, the oral amoxicillin [40 mg/(kg.d), tid] was used for 10 days. Totally 236 cases aged from 0.5 to 12 years were enrolled and 212 cases completed the study. These patients were followed up twice and clinical signs and symptoms were recorded, otoscopy, peripheral blood WBC count, hearing test (pure tone test) and tympanography were performed. RESULTS: In the ceftriaxone group, 103/106 cases were cured or improved (97.17%), while in the amoxicillin group 96/106 cases were cured or improved (90.57%) (P < 0.05). Ceftriaxone was significantly better than amoxicillin in the treatment. Totally 4 cases had side effects such as papular skin rash, urticaria around mouth, skin pigmentation, two cases in the ceftriaxone group and other two cases in the amoxicillin group. There was no significant difference between the 2 groups in side effects. CONCLUSION: Ceftriaxone injection was significantly better than ten-day oral amoxicillin for treatment of acute otitis media in children. The single dose regimen with ceftriaxone seems to be a good choice for children, particularly for. PMID- 14759322 TI - [Dynamic changes of plasma D-dimer in children with traumatic cerebral infarction]. PMID- 14759326 TI - [Changes of C-type natriuretic peptide and neurotensin in rabbits brain injury induced by endotoxin]. PMID- 14759327 TI - [Effects of adriamycin on cultured mouse podocytes VEGF expressions and the potential protective effects of dexamethasone]. PMID- 14759328 TI - [Determination and clinical evaluation of serum procalcitonin in children with pneumonia]. PMID- 14759329 TI - [Basal levels for gonadotropins in 318 children from birth to 6 years of age]. PMID- 14759330 TI - [Correlation analysis of CD40L expression of peripheral T lymphocytes and serum level of IgE in children with allergic asthma]. PMID- 14759331 TI - [Allogenic hematopoietic stem cell transplantation complicated with implantation syndrome in 4 cases]. PMID- 14759332 TI - [The investigation of pathologic oxygen supply dependency]. PMID- 14759333 TI - [Antiphospholipid syndrome]. PMID- 14759335 TI - [Hypertensive encephalopathy caused by transfusing back of frozen cord blood hematopoietic cells caused in one case]. PMID- 14759336 TI - Current topics in herpes research. PMID- 14759337 TI - Herpesvirus infections in transplant recipients: current challenges in the clinical management of cytomegalovirus and Epstein-Barr virus infections. AB - Herpesvirus infections are common causes of morbidity and mortality in solid organ and haematopoietic stem cell transplant recipients. Significant scientific breakthroughs have reduced the incidence of herpesvirus infections during the early post-transplant period, but herpesvirus infections continue to impact significantly on the outcome of transplantation. Widespread and prolonged use of antiviral drugs has not eliminated the risk of cytomegalovirus (CMV) disease, but has changed its natural course by delaying its onset. Moreover, there is now increased recognition of ganciclovir (and other antiviral drug)-resistant CMV infections. By contrast, optimal preventive and treatment strategies for Epstein Barr virus-related post-transplant lymphoproliferative disorders (PTLD) remain elusive. Recent experimental and clinical data, however, demonstrate a promising role for immunotherapy in preventing and treating PTLD. This review highlights the current challenges in managing CMV and Epstein-Barr virus infections in transplant patients. PMID- 14759338 TI - The pharmacokinetics of antiviral therapy in paediatric patients. AB - Effective treatment requires administration of optimal drug dose. In paediatric patients this may be difficult to achieve due to lack of pharmacokinetic studies, administration of small doses, changes in body composition, continuous growth, and development and maturation of organs and systems. This article reviews the factors and misconceptions associated with determining optimal drug dose and route of administration in different paediatric populations. The pharmacokinetics, appropriate doses and preferred routes of administration for aciclovir, ganciclovir, famciclovir, foscarnet and cidofovir, for treating herpesvirus infections in the paediatric population, are discussed. PMID- 14759339 TI - Kaposi's sarcoma-associated herpesvirus (KSHV)/human herpesvirus 8 (HHV-8) as a tumour virus. AB - Kaposi's sarcoma-associated herpesvirus (KSHV) or human herpesvirus 8 (HHV-8) is present in all clinical variants of Kaposi's sarcoma and two lymphoproliferative diseases: primary effusion lymphoma and multicentric Castleman's disease (MCD). Seroepidemiological studies have proved the link between KSHV/HHV-8 infection and development of Kaposi's sarcoma. The virus encodes a number of genes homologous to human genes involved in cell proliferation, anti-apoptosis, angiogenesis and cytokine action. In Kaposi's sarcoma and primary effusion lymphoma, however, most tumour cells only show evidence of latent infection, with a restricted gene expression pattern including latency associated nuclear antigen 1, able to interact with p53 and the retinoblastoma gene. The small number of tumour cells expressing lytic genes may, however, be important for tumour growth by paracrine mechanisms as has been shown for the viral G-protein-coupled receptor. Understanding these molecular mechanisms would provide the rationale for new treatments and prevention of KSHV/HHV-8-associated diseases. PMID- 14759340 TI - Epstein-Barr virus-associated cancers: aetiology and treatment. AB - Evidence implicates Epstein-Barr virus (EBV) in the pathogenesis of tumours arising in lymphoid or epithelial tissue. EBV may adopt different forms of latent infection in different tumour types, reflecting the complex interplay between virus and host-cell environment. Immune response is another important factor influencing EBV gene expression, and viral latent proteins to which immunodominant CTL responses are directed (namely the EBNA3 family), are downregulated in virus-associated tumours arising in overtly immunocompetent individuals. Understanding the functions of EBV latent proteins will improve our knowledge of the role of the virus in transformation, and may help to identify novel therapies. PMID- 14759341 TI - Post-Polio Syndrome. AB - Post-polio syndrome (PPS) is the term used for the new late manifestations that occur in patients 30 to 40 years after the occurrence of acute poliomyelitis. PPS has been recognized for over 100 years, but is more common at the present time because of the large epidemics of poliomyelitis in the 1940s and 1950s. PPS is manifested by neurologic, musculoskeletal, and general manifestations. Neurologic manifestations include new weakness, muscle atrophy, dysphagia, dysphonia, and respiratory failure. Musculoskeletal manifestations include muscle pain, joint pain, spinal spondylosis and scoliosis, and secondary root and peripheral nerve compression. General manifestations include generalized fatigue and cold intolerance. New muscle weakness of a mild-to-moderate degree responds well to a nonfatiguing exercise program and pacing of activity with rest periods to avoid muscle overuse. Generalized fatigue may be treated with energy conservation and weight loss programs and lower extremity orthoses. Pharmacologic agents also may be helpful, but have not been beneficial in controlled trials. Bulbar muscle weakness includes dysphagia, dysphonia, sleep disorders, and chronic respiratory failure. Dysphagia may be improved with instruction on compensatory swallowing techniques. Dysphonia is treated with voice exercise therapy and voice amplification devices. Sleep disorders are treated similarly to sleep disorders in non-PPS patients. Respiratory failure may be treated with continuous positive airway pressure, bilevel positive airway pressure, and nasal ventilation, or tracheotomy and permanent ventilation if necessary. Musculoskeletal (muscle and joint) pain is treated with weight loss, pacing of activities, use of assistive devices, and prescribing anti-inflammatory medications and physical therapy techniques. Cardiopulmonary conditioning can be improved without muscle overuse with cycle or arm ergometer exercise or dynamic aquatic exercise. PMID- 14759342 TI - Leprosy (Hansen's Disease). AB - Leprosy (Hansen's disease) causes the most common treatable form of neuropathy in the world. Several endemic countries account for the majority of the world's cases and most of the cases seen in the US are amongst immigrants. However, endemic cases of leprosy occur in the US. The pathogen is Mycobacterium leprae, a slow-growing, obligate intracellular pathogen that consistently infects skin and peripheral nerves. The clinical appearance of the skin and neurologic deficits develop months to years after infection and are determined by the host's response to the infection. An individual's disease classification can change over time based on the immune status of the individual. Immune-mediated "reactional states" may also occur that require additional recognition and treatment. Varied in its manifestations, a successful treatment approach relies on proper recognition and classification of disease. PMID- 14759344 TI - Marine Neurotoxins: Envenomations and Contact Toxins. AB - Familiarity with the appearance and habitat of venomous sea creatures, the location of their stinging apparatus, and surveillance of population concentrations within recreational waters are essential in avoiding envenomations. Compared with the thermo-stable low molecular weighted ingestible seafood toxins, venomous toxins are often large molecular weight proteins and many are heat labile, which provides opportunity for therapeutic intervention. Heat therapy may denature the toxins, and provide immediate relief of pain in coelenterate and venomous fish envenomations. Injections of local anesthetic agents may also be used. First aid measures at the seashore may limit the spread of venom, and include immobilization of the affected sites, compression bandaging, and venous-lymphatic occlusive bandages. Measures to limit continued envenomation by attached stinging cells include topical vinegar for jellyfish tentacles and irrigation with debridment for spines of venomous fish. Antivenins are of limited availability and may be used for envenomations with sea snakes, Chironex box jellyfish, and some venomous fish. Sea snakes bites may be treated with antivenin from land snakes or with hemodialysis when antivenin is not available. Neuromuscular paralysis occurs with bites by sea snakes, cone snails, blue octopuses, and some jellyfish. Supportive treatment includes attention to cardiopulmonary resuscitation and intubation. Exposure to Pfeisteria may result in cognitive and behavioral abnormalities. Treatment with cholestyramine may be helpful in binding the toxin and improve recovery. PMID- 14759343 TI - Marine Neurotoxins: Ingestible Toxins. AB - Fish and shellfish account for a significant portion of food-borne illnesses throughout the world. In general, three classes of diseases result from seafood consumption--intoxication, allergies, and infections. In this review, the authors discuss several seafood-borne toxins, including domoic acid, which acts on the central nervous system. In addition, the authors discuss ciguatoxin-, brevetoxin , saxitoxin-, tetrodotoxin-, and scombroid-related histamine toxicity, all of which act primarily on the peripheral nervous system. Fish has become a very popular food in the US mostly related to its potential health benefits. Fish is consumed to such a degree that fishing stocks are reportedly at an all time low from what seemed like an endless supply even 30 years ago. One of the most significant threats to human intoxication is the recreational harvest of shellfish, often times located in remote locations where the harvesters are subsistent on fishery resources and have no monitoring in place. The hazard to intoxication is not as common in purchased seafood, which is more stringently regulated, yet still is a serious problem. Most ingestible toxins are thermo stable and therefore unaffected by cooking, freezing, or salting. Air transport of consumable products throughout the world makes it easy to obtain exotic edibles from far away countries. A seemingly unusual toxin can be more commonly encountered than previously thought and it is important to consider this when evaluating patients. Recognition and treatment of various neurologic symptoms related to seafood ingestion is paramount in today's mobile, gastronomic world. Specific treatments vary with each individual toxin and with the individual's specific reaction to the toxin. Generally, some degree of medical care is required with all ingestible toxin exposure, ranging from simple administration of medication and hydration to ventilatory and cardiovascular support. PMID- 14759345 TI - Cerebral Malaria. AB - Successful management of cerebral malaria requires rapid diagnosis, initiation of appropriate antimalarial agents, and aggressive supportive care. The antimalarial used will depend on drug availability, with many of the most effective agents not routinely available in the US. The use of technologically advanced diagnostic studies, such as electroencephalogram, that are generally not available in malaria-endemic regions may aid in management. But other tests, such as advanced neuroimaging, may offer limited guidance because there are little data indicating the best management of conditions, such as increased intracranial pressure, focal edema, and microhemorrhages. Close clinical assessments and rapid initiation of proven therapies should be the focus of care, even when advanced resources are available. Given the mortality of this condition and the long-term neurologic morbidity seen in survivors, adjunctive therapies are clearly needed to improve patient outcomes. Until more data are available, much of the management of cerebral malaria will continue to be based on anecdote rather than evidence. PMID- 14759346 TI - HIV Dementia. AB - Dementia and milder forms of cognitive impairment are important manifestations, with important socioeconomic consequences, of HIV infection. Early detection and aggressive treatment are essential to achieve stabilization and prevent progression. In some affected individuals, reversal of symptoms may follow initiation of effective therapy. Antiretroviral treatment needs to be individualized, taking into consideration the relative central nervous system penetration of the drugs, presence of drug resistant viral strains, possible drug interactions, and drug side effect profiles. Combination antiretroviral therapy is preferred. Zidovudine, stavudine, abacavir, navirapine and indinavir have relatively good penetration into the central nervous system. The aim of therapy should be for control of viral load in the periphery and the cerebrospinal fluid. Furthermore, although not established therapy, the use of neuroprotective agents and anti-inflammatory regimens may be considered. A comprehensive multidisciplinary approach to management with regular follow-up is essential for treatment. PMID- 14759347 TI - Inflammatory Myopathies. AB - Therapies that suppress or modify the immune system remain the primary treatment for the idiopathic inflammatory myopathies. Dermatomyositis (DM) and polymyositis (PM) are the two conditions that respond best to immunotherapy. Although there are no randomized controlled trials, corticosteroids, specifically high-dose oral prednisone, remain the cornerstone of management. Recent controlled clinical trials show that intravenous immunoglobulin (IVIg) is an efficacious treatment in DM. Expert clinicians are generally using this as a second-line agent or as an adjunct to prednisone. IVIg has a relatively benign side effect profile compared with chronic steroid use, but the cost of treatment, the need for repetitive treatment cycles, and the potential for serious adverse effects have kept it from being a first-line agent in DM. There have been no trials performed using IVIg in PM. Chronic immunosuppressant medications, including azathioprine, cyclosporine, and methotrexate, are also available for long-term management in patients with recalcitrant disease or side effects from extended corticosteroid use. These agents lack the troubling side effects of prednisone and are less costly than IVIg, but require close medical monitoring for adverse reactions to blood, kidney, lung, or liver. Newer medications with potentially more benign side effect profiles, such as mycophenolate mofetil and etanercept, are currently being studied, but knowledge of how effective they are and how quickly they work are not yet available. Inclusion body myositis has proven resistant to immunosuppressive medications. The response has been so consistently poor and so easily contrasted with DM that the authors wonder why these conditions are so routinely lumped together in chapters and review articles. Clearly, this is based solely on the common pathologic feature of inflammation, rather than a clear understanding of how these conditions occur, or why they do or do not respond to treatment. PMID- 14759348 TI - Myasthenia Gravis. AB - Myasthenia gravis is a disorder of neuromuscular transmission, manifest by variable weakness of skeletal muscle. The disorder has a range of therapies that differ with respect to efficacy, timing, and side effects. The physician treating myasthenia gravis must be well versed in understanding the evidence basis for using these agents, as well as the trade-offs between persistent disease manifestations and their costs and expected benefits. Diagnosis of myasthenia gravis depends on recognizing the pattern of weakness, which typically involves some combination of extraocular, bulbar, facial, limb, and neck muscles. Management relies on some combination of medications that influence the function of the neuromuscular junction and treatments that alter the immune response. Thymectomy is commonly used, although trends in evidence-based medicine are leading expert clinicians to look closely at its efficacy. Plasma exchange is useful for patients in crisis, who require rapid improvement. The exact role for high-dose intravenous immunoglobulin in this setting is still being studied, although the agent is gaining popularity. Knowing that the treatments are effective is not enough. It is still important to determine the treatment that has a more rapid onset, because these patients often require intensive care or respiratory assistance. PMID- 14759349 TI - My cardiovascular wish list for 2004. PMID- 14759350 TI - Aortic valve disease: current recommendations. AB - Aortic stenosis is a common valvular abnormality. Surgical valve replacement is the standard of care for symptomatic patients with severe aortic stenosis and is appropriate for certain asymptomatic patients. Statin therapy may slow progression of the disease. Contrary to conventional wisdom, vasodilator therapy may in fact be safe and effective in certain instances of severe aortic stenosis. Chronic aortic regurgitation is commonly treated with vasodilator therapy, which is certainly indicated for the asymptomatic patient with hypertension. Surgery is recommended for left ventricular dilatation, diminished ventricular systolic function, or symptoms. PMID- 14759351 TI - Mitral regurgitation in chronic heart failure: more questions than answers? AB - The presence, implications, and approach to mitral valvular disease in the setting of a cardiomyopathy has recently become a focal issue for discussion. Although recent evidence suggests that mitral regurgitation confers a poor prognosis in heart failure, the true prevalence of mitral regurgitation as well as its pathogenic contribution to prognosis in heart failure remains uncertain. Whereas angiotensin-converting enzyme inhibitors have not been demonstrated to favorably alter mitral regurgitation, treatment with b-blockers or cardiac resynchronization therapy has been shown to induce regression of ventricular dilation, with consequent decrease in mitral regurgitation. Surgical valve reconstruction by undersized ring annuloplasty appears promising, but optimal results require skilled surgical expertise. Even in highly experienced hands, intermediate outcomes appear to be inferior to those of cardiac transplantation. Percutaneous mitral valve repair remains an investigational procedure that warrants further clinical research. PMID- 14759352 TI - Newer concepts in the surgical treatment of valvular heart disease. AB - The search for the ideal therapy for valve replacement continues. The major options include mechanical or tissue valves, with an increasing variety of tissue valves becoming available. The key factor continues to be thrombogenicity versus durability. Aortic valve surgery primarily consists of replacement. This is aided by the increased variety of options allowing tailoring of the procedure to the patient's native valve disease. Mitral valve surgery has greater potential for repair, which affords preservation of the native valve, optimizing function and reducing long-term complications. An increasingly popular concept is treatment of secondary or functional mitral valve regurgitation in the setting of depressed left ventricular function. The routine use of intraoperative transesophageal echocardiography and a trend toward the use of minimally invasive procedures are altering the conduct of valve operations. PMID- 14759353 TI - Complications of prosthetic heart valves. AB - Treatment of native valvular heart disease has resulted in an increasing number of patients with prosthetic valves. Although an improvement over the diseased native valve removed at surgery, prosthetic valves have suboptimal hemodynamics; mechanical valves require anticoagulation and tissue valves wear out over time. Serious complications of prosthetic valves occur at a rate of about 2% to 3% per patient-year. Complications include thromboembolism, prosthesis-patient mismatch, structural valve dysfunction, endocarditis, and hemolysis. Prosthetic valve endocarditis is a lethal disease with mortality rates of 50% to 80% even with appropriate therapy. Echocardiography now provides detailed information on valve function and hemodynamics, allowing early detection of complications. Many of these complications can be prevented by choosing the optimal valve at the time of surgery, rigorous control of anticoagulation and adherence to established anticoagulation guidelines, dental hygiene and endocarditis prophylaxis, and periodic echocardiographic monitoring by a cardiologist. PMID- 14759354 TI - The African American antiplatelet stroke prevention study. PMID- 14759355 TI - The losartan intervention for endpoint reduction substudy. PMID- 14759356 TI - Mechanisms of ischemic brain injury. AB - Stroke is the third leading cause of death and the leading cause of long-term disability in the United States. Approximately 80% of all strokes are ischemic and there are limited therapies approved for the treatment of acute ischemic stroke. Understanding the mechanisms of ischemic brain damage is necessary for the development of innovative treatment strategies. In this review, we discuss the hemodynamic and molecular mechanisms of ischemic brain damage and the potential therapeutic strategies, including reperfusion and primary and secondary neuroprotection, and strategies for recovery of function, such as neural plasticity and stem cell transplantation. The effective treatment of ischemic stroke is likely to result from a combination of therapeutic modalities aimed at different mechanisms of ischemic brain damage and delivered at specific times after acute cerebral ischemia. PMID- 14759357 TI - Treatment of hypertension and prevention of ischemic stroke. AB - Data clearly indicate that treatment with antihypertensive drugs reduces the incidence of all strokes in men (by 34%), women (by 38%), elderly persons (by 36%), including those older than 80 years (by 34%), younger persons, those with systolic and diastolic hypertension, persons with isolated systolic hypertension, and in those with a history of stroke or transient ischemic attack (by 28%). Blood pressure should be reduced to less than 140/90 mm Hg. The overall data also suggest that reduction of stroke in persons with hypertension is related more to a reduction in blood pressure than to the type of antihypertensive drugs used. PMID- 14759358 TI - Antiphospholipid antibodies and stroke. AB - Antiphospholipid antibodies are associated with venous and arterial thrombo occlusive events, including stroke. They are a heterogeneous family of antibodies that react to negatively charged membrane-bound phospholipids or phospholipid protein complexes. Despite much progress in the past decade, the precise mechanisms by which they promote thrombosis remain unknown. Increased risk of initial, but perhaps not recurrent, stroke has been associated with their presence. Antithrombotic regimens have been the mainstay of treatment, but the ideal therapeutic approach remains to be defined. This paper reviews the history, definition, associations, and treatment of this complex entity. PMID- 14759359 TI - The role of warfarin and aspirin in secondary prevention of stroke. AB - Antithrombotic therapy plays a central role in secondary prevention after ischemic stroke and transient ischemic attack. The choice among warfarin, aspirin, and other antiplatelet agents, however, depends on the cause of stroke and other individual patient characteristics. The use of warfarin anticoagulation in patients with atrial fibrillation and ischemic stroke has demonstrated robust reductions in risk of recurrent events, comparable with those achieved in primary prevention. Warfarin may also be recommended for patients with other high-risk cardioembolic sources of stroke. The role of warfarin in noncardioembolic ischemic stroke is more controversial. The Warfarin Aspirin Recurrent Stroke Study found no evidence of superiority of warfarin over aspirin in stroke patients overall, nor in any major stroke subtype, including those patients with patent foramen ovale. In post-hoc analyses, there was some evidence of benefit with warfarin in patients with cryptogenic stroke without hypertension. Risks of major bleeding did not differ significantly between warfarin and aspirin groups. For most patients with noncardioembolic strokes, therefore, antiplatelet therapy is the preferred option, although clinician experience still dictates practice in individual situations. Newer antiplatelet agents, and the combination of novel agents with aspirin, are also finding a role in stroke prevention as clinical trial data become available. PMID- 14759360 TI - Management of patients with stroke and a patent foramen ovale. AB - Observational studies have established a strong association between the presence of patent foramen ovale (PFO) and increased risk of ischemic stroke. The mechanism involved is presumed to be a paradoxical embolism from a venous thrombus that travels via the PFO to the systemic circulation. The best treatment modality to prevent recurrent stroke in patients with PFO remains undefined. There are four major choices: 1) medical therapy with anticoagulants, 2) medical therapy with antiplatelet agents, 3) surgical closure, and 4) percutaneous device closure. The Patent Foramen Ovale in Cryptogenic Stroke Study has demonstrated that antiplatelet and anticoagulant therapies are of equal benefit in preventing recurrent neurologic events in stroke patients with a PFO. Medical therapy should remain as the initial choice of secondary prophylactic therapy. PFO closure, either surgical or percutaneous, may further reduce event rates; however, this remains to be demonstrated because no randomized trial to date has compared PFO closure with medical therapy. PMID- 14759362 TI - Reprogrammed genetic decoding in cellular gene expression. AB - Reprogrammed genetic decoding signals in mRNAs productively overwrite the normal decoding rules of translation. These "recoding" signals are associated with sites of programmed ribosomal frameshifting, hopping, termination codon suppression, and the incorporation of the unusual amino acids selenocysteine and pyrrolysine. This review summarizes current knowledge of the structure and function of recoding signals in cellular genes, the biological importance of recoding in gene regulation, and ways to identify new recoded genes. PMID- 14759363 TI - The structural basis for autoinhibition of FLT3 by the juxtamembrane domain. AB - FLT3 is a type III receptor tyrosine kinase that is thought to play a key role in hematopoiesis. Certain classes of FLT3 mutations cause constitutively activated forms of the receptor that are found in significant numbers of patients with acute myelogenous leukemia (AML). The mutations occur either in the activation loop, for example, as point mutations of Asp835 or as internal tandem duplication (ITD) sequences in the juxtamembrane (JM) domain. To further understand the nature of FLT3 autoinhibition and regulation, we have determined the crystal structure of the autoinhibited form of FLT3. This structure shows the autoinhibitory conformation of a complete JM domain in this class of receptor tyrosine kinases. The detailed inhibitory mechanism of the JM domain is revealed, which is likely utilized by other members of type III receptor tyrosine kinases. PMID- 14759364 TI - HIV-1 Nef mimics an integrin receptor signal that recruits the polycomb group protein Eed to the plasma membrane. AB - The Nef protein of human and simian immunodeficiency virus (HIV/SIV) is believed to interfere with T cell activation signals by forming a signaling complex at the plasma membrane. Composition and function of the complex are not fully understood. Here we report that Nef recruits the Polycomb Group (PcG) protein Eed, so far known as a nuclear factor and repressor of transcription, to the membrane of cells. The Nef-induced translocation of Eed led to a potent stimulation of Tat-dependent HIV transcription, implying that Eed removal from the nucleus is required for optimal Tat function. Similar to Nef action, activation of integrin receptors recruited Eed to the plasma membrane, also leading to enhanced Tat/Nef-mediated transcription. Our results suggest a link between membrane-associated activation processes and transcriptional derepression and demonstrate how HIV exploits this mechanism. PMID- 14759365 TI - Kinetic determinants of high-fidelity tRNA discrimination on the ribosome. AB - The ribosome selects aminoacyl-tRNA (aa-tRNA) matching to the mRNA codon from the bulk of non-matching aa-tRNAs in two consecutive selection steps, initial selection and proofreading. Here we report the kinetic analysis of selection taking place under conditions where the overall selectivity was close to values observed in vivo and initial selection and proofreading contributed about equally. Comparison of the rate constants shows that the 350-fold difference in stabilities of cognate and near-cognate codon-anticodon complexes is not used for tRNA selection due to high rate of GTP hydrolysis in the cognate complex. tRNA selection at the initial selection step is entirely kinetically controlled and is due to much faster (650-fold) GTP hydrolysis of cognate compared to near-cognate substrate. PMID- 14759366 TI - The Glc7p nuclear phosphatase promotes mRNA export by facilitating association of Mex67p with mRNA. AB - mRNA export is mediated by Mex67p:Mtr2p/NXF1:p15, a conserved heterodimeric export receptor that is thought to bind mRNAs through the RNA binding adaptor protein Yra1p/REF. Recently, mammalian SR (serine/arginine-rich) proteins were shown to act as alternative adaptors for NXF1-dependent mRNA export. Npl3p is an SR-like protein required for mRNA export in S. cerevisiae. Like mammalian SR proteins, Npl3p is serine-phosphorylated by a cytoplasmic kinase. Here we report that this phosphorylation of Npl3p is required for efficient mRNA export. We further show that the mRNA-associated fraction of Npl3p is unphosphorylated, implying a subsequent nuclear dephosphorylation event. We present evidence that the essential, nuclear phosphatase Glc7p promotes dephosphorylation of Npl3p in vivo and that nuclear dephosphorylation of Npl3p is required for mRNA export. Specifically, recruitment of Mex67p to mRNA is Glc7p dependent. We propose a model whereby a cycle of cytoplasmic phosphorylation and nuclear dephosphorylation of shuttling SR adaptor proteins regulates Mex67p:Mtr2p/NXF1:p15-dependent mRNA export. PMID- 14759367 TI - The doubletime and CKII kinases collaborate to potentiate Drosophila PER transcriptional repressor activity. AB - In all genetically studied model organisms, a negative feedback loop of gene expression contributes to the circadian rhythm mechanism. In the Drosophila system, it has been proposed that the delay between the synthesis and function of clock proteins is due to phosphorylation-regulated nuclear entry. To test this hypothesis, we assayed the relationship between PER phosphorylation, nuclear localization, and transcriptional repression activity in cultured S2 cells. The results indicate that the two putative PER kinases DBT and CKII work together to phosphorylate PER and increase repression activity. Experiments combining kinase inhibition with inhibition of PER nuclear export suggest that phosphorylation directly affects PER repression activity and that PER nuclear localization is an indirect consequence of the association of active PER with DNA or chromatin. This interpretation suggests further that the circadian regulation of PER nuclear localization in flies reflects changes in PER transcriptional activity rather than in PER nuclear import or export activity. PMID- 14759368 TI - High-definition macromolecular composition of yeast RNA-processing complexes. AB - A remarkably large collection of evolutionarily conserved proteins has been implicated in processing of noncoding RNAs and biogenesis of ribonucleoproteins. To better define the physical and functional relationships among these proteins and their cognate RNAs, we performed 165 highly stringent affinity purifications of known or predicted RNA-related proteins from Saccharomyces cerevisiae. We systematically identified and estimated the relative abundance of stably associated polypeptides and RNA species using a combination of gel densitometry, protein mass spectrometry, and oligonucleotide microarray hybridization. Ninety two discrete proteins or protein complexes were identified comprising 489 different polypeptides, many associated with one or more specific RNA molecules. Some of the pre-rRNA-processing complexes that were obtained are discrete sub complexes of those previously described. Among these, we identified the IPI complex required for proper processing of the ITS2 region of the ribosomal RNA primary transcript. This study provides a high-resolution overview of the modular topology of noncoding RNA-processing machinery. PMID- 14759369 TI - Ras induces mediator complex exchange on C/EBP beta. AB - C/EBPbeta is an intrinsically repressed transcription factor that regulates genes involved in differentiation, proliferation, tumorigenesis, and apoptosis. C/EBPbeta acts as a repressor that is turned into an activator by the Ras oncoprotein through phosphorylation of a MAPK site. C/EBPbeta activation is accompanied by a conformational change. Active and repressive C/EBPbeta interacts with multisubunit Mediator complexes through the CRSP130/Sur2 subunit. The CRSP130/Sur2 subunit is common to two distinct types of Mediator complexes, characterized by CRSP70 and CDK8 proteins as transcriptionally active and inactive Mediator, respectively. Knockdown of CRSP130/Sur2 prevents Mediator binding and transactivation through C/EBPbeta. Oncogenic Ras signaling or activating mutations in C/EBPbeta selects the transcriptionally active Mediator complex that also associates with RNA polymerase II. These results show that a Ras-induced structural alteration of C/EBPbeta determines differential gene activation through selective interaction with distinct Mediator complexes. PMID- 14759370 TI - Structural mechanism of the bromodomain of the coactivator CBP in p53 transcriptional activation. AB - Lysine acetylation of the tumor suppressor protein p53 in response to a wide variety of cellular stress signals is required for its activation as a transcription factor that regulates cell cycle arrest, senescence, or apoptosis. Here, we report that the conserved bromo-domain of the transcriptional coactivator CBP (CREB binding protein) binds specifically to p53 at the C terminal acetylated lysine 382. This bromodomain/acetyl-lysine binding is responsible for p53 acetylation-dependent coactivator recruitment after DNA damage, a step essential for p53-induced transcriptional activation of the cyclin dependent kinase inhibitor p21 in G1 cell cycle arrest. We further present the three-dimensional nuclear magnetic resonance structure of the CBP bromodomain in complex with a lysine 382-acetylated p53 peptide. Using structural and biochemical analyses, we define the molecular determinants for the specificity of this molecular recognition. PMID- 14759371 TI - The histone-fold protein complex CHRAC-15/17 enhances nucleosome sliding and assembly mediated by ACF. AB - The histone fold is a structural motif with which two related proteins interact and is found in complexes involved in wrapping DNA, the nucleosome, and transcriptional regulation, as in NC2. We reveal a novel function for histone fold proteins: facilitation of nucleosome remodeling. ACF1-ISWI complex (ATP dependent chromatin assembly and remodeling factor [ACF]) associates with histone fold proteins (CHRAC-15 and CHRAC-17 in the human chromatin accessibility complex [CHRAC]) whose functional relevance has been unclear. We show that these histone fold proteins facilitate ATP-dependent nucleosome sliding by ACF. Direct interaction of the CHRAC-15/17 complex with the ACF1 subunit is essential for this process. CHRAC-17 interacts with another histone-fold protein, p12, in DNA polymerase epsilon, but CHRAC-15 is essential for interaction with ACF and enhancement of nucleosome sliding. Surprisingly, CHRAC-15/17, p12/CHRAC-17, and NC2 complexes facilitate ACF-mediated chromatin assembly by a mechanism different from nucleosome sliding enhancement, suggesting a general activity of H2A/H2B type histone-fold complexes in chromatin assembly. PMID- 14759372 TI - Healing the wounds inflicted by sleeping beauty transposition by double-strand break repair in mammalian somatic cells. AB - The Sleeping Beauty (SB) element is a useful tool to probe transposon-host interactions in vertebrates. We investigated requirements of DNA repair factors for SB transposition in mammalian cells. Factors of nonhomologous end joining (NHEJ), including Ku, DNA-PKcs, and Xrcc4 as well as Xrcc3/Rad51C, a complex that functions during homologous recombination, are required for efficient transposition. NHEJ plays a dominant role in repair of transposon excision sites in somatic cells. Artemis is dispensable for transposition, consistent with the lack of a hairpin structure at excision sites. Ku physically interacts with the SB transposase. DNA-PKcs is a limiting factor for transposition and, in addition to repair, has a function in transposition that is independent from its kinase activity. ATM is involved in excision site repair and affects transposition rates. The overlapping but distinct roles of repair factors in transposition and in V(D)J recombination might influence the outcomes of these mechanistically similar processes. PMID- 14759373 TI - CTCF tethers an insulator to subnuclear sites, suggesting shared insulator mechanisms across species. AB - Insulators can block an enhancer of one gene from activating a promoter on another nearby gene. Almost all described vertebrate insulators require binding of the regulatory protein CTCF for their activity. We show that CTCF copurifies with the nucleolar protein nucleophosmin and both are present at insulator sites in vivo. Furthermore, exogenous insulator sequences are tethered to the nucleolus in a CTCF-dependent manner. These interactions, quite different from those of the gypsy insulator element in Drosophila, may generate similar loop structures, suggesting a common theme and model for enhancer-blocking insulator action. PMID- 14759374 TI - Prognostic usefulness of marginal troponin T elevation. AB - Marginal elevations of troponin T among patients with chest pain are often considered to be insignificant. We sought to define the prognostic value of marginal troponin T elevations in patients presenting to the emergency department with suspected myocardial ischemia. Four hundred twenty-eight consecutive patients presenting to the emergency department with ongoing chest pain were evaluated, followed through their hospital course, and contacted for follow-up 4 months after discharge. Two hundred ninety-nine patients had undetectable troponin T levels (<0.01 microg/L), 76 had marginal troponin T elevations (0.01 to 0.09 microg/L), and 53 had frank troponin T elevations (> or =0.1 microg/L). Patients with either marginally or frank elevated troponin levels were older and more likely to be men, but did not differ from patients with undetectable troponin levels with regard to the prevalence of coronary artery disease risk factors, history of coronary disease, or race. While in the hospital, the undetectable and marginal troponin groups were referred for cardiac testing in equal proportions (58% and 59%, respectively), whereas 87% of the elevated group underwent further testing. After adjustment for possible confounders, a significantly increased rate of death/myocardial infarction/revascularization was observed in the marginal troponin group compared with the undetectable troponin group (p = 0.004). Marginal elevations of troponin T identified a currently underevaluated high-risk subgroup of patients with suspected myocardial ischemia who are more likely to have adverse clinical outcomes than those with undetectable troponin levels. PMID- 14759375 TI - Comparison of reperfusion regimens with or without tirofiban in ST-elevation acute myocardial infarction. AB - There is continued debate as to whether a combined reperfusion regimen with platelet glycoprotein IIb/IIIa inhibitors provides additional benefit in optimal myocardial reperfusion of patients with a ST-elevation acute myocardial infarction (AMI). In addition, the best angiographic method to evaluate optimal myocardial reperfusion is still controversial. Patients (n = 144) with a first AMI presenting <6 hours from onset of symptoms were randomized to receive a conjunctive strategy (n = 72) with low-dose alteplase (50 mg) and tirofiban (0.4 microg/kg/min/30 minute bolus; infusion of 0.1 microg/kg/minute), or tirofiban plus stenting percutaneous coronary intervention (PCI). Control patients (n = 72) received standard strategy with either full-dose alteplase (100 mg) or stenting PCI [correction]. All patients were submitted to coronary angiographic study at 90 minutes. The primary end point was Thrombolysis In Myocardial Infarction (TIMI) grade 3 flow at 90 minutes. Secondary end points were TIMI myocardial perfusion (TMP) rates, a composite end point at 30 days (death, reinfarction, refractory ischemia, stroke, heart failure, revascularization procedures, or pulmonary edema), and bleeding or hematologic variables. The rate of TIMI 3 flow at 90 minutes for patients treated with alteplase alone was 42% compared with 64% for those who received low-dose alteplase and tirofiban. Standard stenting PCI achieved 81% of TIMI 3 flow compared with 92% when tirofiban was used. Significantly higher rates of TMP grade 3 were observed when tirofiban was used as the adjunctive treatment in both alteplase (66% vs 47%) and stenting PCI (73% vs 55%). Higher rates of the composite end point were observed in standard regimens compared with conjunctive regimens (hazard ratio 5.8, 95% confidence interval 1.27 to 26.6, p = 0.023). Regardless of reperfusion regimen, better outcomes were observed when a combination of TIMI 3 flow and TMP grade 3 was achieved. Beyond TIMI 3 flow rate, the TMP grade was an important determinant. The rates of major bleeding were similar (2.8%) for standard versus conjunctive regimens with tirofiban. Thus, tirofiban as a conjunctive therapy for lytic and stenting regimens not only improves TIMI 3 flow rates, but also the TMP3 rates, which are related to a better clinical outcome without an increase in the risk of major bleeding. This study supports the hypothesis that platelets play a key role not only in the atherothrombosis process, but also in the disturbances of microcirculation and tissue perfusion. PMID- 14759376 TI - Six-month outcomes in a multinational registry of patients hospitalized with an acute coronary syndrome (the Global Registry of Acute Coronary Events [GRACE]). AB - Relatively limited data are available, particularly from the perspective of a multinational registry, about the post-discharge outcomes and management practices of patients with an acute coronary syndrome (ACS). The objectives of this longitudinal study were to examine 6-month outcomes in a large multinational sample of patients hospitalized with an ACS. A total of 5,476 patients with ST segment elevation acute myocardial infarction (STEAMI), 5,209 patients with non ST-segment elevation acute myocardial infarction (NSTEAMI), and 6,149 patients with unstable angina pectoris discharged from 90 hospitals in 14 countries comprised the study population. The study sample was recruited from 18 cluster sites in 14 countries that are currently collaborating in the Global Registry of Acute Coronary Events (GRACE) study. The 6-month post-discharge death rates were 4.8% in patients with STEAMI, 6.2% in patients with NSTEAMI, and 3.6% in patients with unstable angina pectoris. Approximately 1 in 5 of each of our comparison groups were rehospitalized for heart disease during the 6-month follow-up, and approximately 15% of each of the respective study cohorts underwent coronary revascularization during follow-up. Demographic and clinical characteristics of post-discharge decedents were identified according to type of ACS. Our results suggest that a considerable proportion of patients who were discharged from the hospital after an ACS, with some differences noted according to type of ACS, remain at increased risk for adverse outcomes during the relatively brief post discharge period. These data suggest the need for better long-term medical management and more intense follow-up of patients with an ACS to improve their long-term outlook. PMID- 14759377 TI - Long-term outcomes of optimized medical management of outpatients with stable coronary artery disease. AB - The objective of this study was to assess long-term clinical outcomes and their correlates in medically managed outpatients with stable angina pectoris, healed myocardial infarction (MI), or documented asymptomatic coronary artery disease (CAD). Management strategy emphasized maximally tolerated medical therapy and modification of coronary risk factors. Referral to invasive coronary interventions followed stricter criteria than standard published guidelines. Primary study outcomes were all-cause mortality or nonfatal myocardial infarction. Secondary study outcomes included cardiac death, unstable angina, or coronary revascularization. A total of 693 men and women with proved CAD (mean age 67 years at entry, 85% men, 41% with history of MI) were enrolled. The annual incidence of nonfatal MI, cardiac mortality, and total mortality was 2.2%, 0.8%, and 1.4%, respectively, during an average follow-up of 4.6 years. Coronary revascularization was performed in 24% of subjects; unstable or progressive anginal symptoms were the most common reasons for revascularization. In patients with documented stable CAD, a management strategy based on intensive medical therapy and modification of established coronary risk factors was associated with excellent long-term outcomes. Thus, coronary interventions can be safely delayed until clinical instability ensues, without increased risk of MI or death. This treatment approach represents a viable alternative to invasive strategies. PMID- 14759378 TI - Value of low-dose dobutamine addition to routine dual isotope gated SPECT myocardial imaging in patients with healed myocardial infarction or abnormal wall thickening by echocardiogram. AB - There is overlap in myocardial viability detection by thallium-201 uptake and contractile reserve (CR) using low-dose dobutamine (LDD). The dual isotope protocol was modified in this study by acquiring thallium-201 images using LDD to enhance viability detection in addition to coronary flow reserve assessment. One hundred twenty-four patients with coronary disease underwent gated single-photon emission computed tomographic thallium-201 imaging at rest with LDD (10 microg/kg/min) during acquisition followed by stress technetium-99m sestamibi myocardial perfusion (MP) imaging with dobutamine, adenosine, or treadmill exercise. F-18-fluorodeoxyglucose (FDG) positron emission tomography was obtained in 41 patients. Myocardial perfusion (MP) imaging was divided into normal, fixed, and ischemic segments, and subclassified by wall motion and/or thickening changes between 1-hour poststress and LDD into normal, fixed, or improved dysfunctional segments (CR present). Mean left ventricular ejection fraction was 39% at 1 hour after stress and 47% with LDD (p <0.001). In dysfunctional myocardium, CR was significantly higher (p <0.001) in ischemic (233 of 368) and fixed segments (150 of 335) than in normal MP segments (43 of 220). Combined MP and CR analysis showed higher accuracy and negative predictive value in identifying FDG-viable myocardium than either method alone, whereas a high positive predictive value was maintained, similar to both markers. Quantitative analysis showed significant increased wall motion and thickening with LDD compared with 1 hour after stress, which was highest in ischemic segments and lowest in fixed segments. Thus, LDD dual isotope is a practical protocol that improves viability detection by simultaneous MP and CR analysis in addition to coronary flow reserve assessment in 1 study. Moreover, it requires no extra imaging time or radioactivity than the routine protocol. PMID- 14759379 TI - Safety and tolerability of simvastatin plus niacin in patients with coronary artery disease and low high-density lipoprotein cholesterol (The HDL Atherosclerosis Treatment Study). AB - The high-density lipoprotein (HDL)-Atherosclerosis Treatment Study showed that simvastatin plus niacin (mean daily dose 13 mg and 2.4 g, respectively) halt angiographic atherosclerosis progression and reduce major clinical events by 60% in patients with coronary artery disease (CAD) who have low HDL, in comparison with placebos, over 3 years. How safe and well-tolerated is this combination? One hundred sixty patients with CAD, including 25 with diabetes mellitus, with mean low-density lipoprotein cholesterol of 128 mg/dl, HDL cholesterol of < or =35 mg/dl (mean 31), and mean triglycerides of 217 mg/dl were randomized to 4 factorial combinations of antioxidant vitamins or their placebos and simvastatin plus niacin or their placebos. Patients were examined monthly or bimonthly for 38 months; side effects (gastrointestinal upset, nausea, anorexia, vision, skin, and energy problems, or muscle aches) were directly queried and recorded. Aspartate aminotransferase, creatine phosphokinase (CPK), uric acid, homocysteine, and fasting glucose levels were regularly monitored. A safety monitor reviewed all side effects and adjusted drug dosages accordingly. Patients who received simvastatin plus niacin and those on placebo had similar frequencies of clinical or laboratory side effects: any degree of flushing (30% vs 23%, p = NS), symptoms of fatigue, nausea, and/or muscle aches (9% vs 5%, p = NS), aspartate aminotransferase (SGOT) > or =3 times upper limit of normal (3% vs 1%, p = NS), CPK > or =2 times upper limit of normal (3% vs 4%, p = NS), CPK > or =5 times upper limit of normal, new onset of uric acid > or =7.5 mg/dl (18% vs 15%, p = NS), and homocysteine > or =15 micromol/L (9% vs 4%, p = NS). Glycemic control among diabetics declined mildly in the simvastatin-niacin group but returned to pretreatment levels at 8 months and remained stable for rest of the study. This combination regimen was repeatedly described by 91% of treated patients and 86% of placebo subjects as "very easy" or "fairly easy" to take. Thus, the simvastatin plus niacin regimen is effective, safe, and well tolerated in patients with or without diabetes mellitus. PMID- 14759380 TI - Usefulness of periprocedural creatinine phosphokinase-MB release to predict adverse outcomes after intracoronary radiation therapy for in-stent restenosis. AB - We aimed to analyze periprocedural creatinine phosphokinase (CPK)-MB elevation in patients treated with intracoronary radiation therapy (IRT) for in-stent restenosis (ISR) to risk stratify these patients. The clinical significance of periprocedural CPK-MB elevation after IRT for ISR is unknown. An elevated CPK-MB has been associated with increased mortality after conventional angioplasty. We evaluated 1,326 patients who were enrolled in radiation trials for ISR at the Washington Hospital Center using gamma- and beta-emitters. Patients were analyzed according to degree of CPK-MB increase within 24 hours of the index IRT procedure (normal CPK-MB, CPK-MB 1 to 3 times the upper limit of normal, or CPK-MB >3 times the upper limit of normal). Patients with CPK-MB >3 times the upper limit of normal were older (64 +/- 12 years, p = 0.04), more likely to be smokers (64%, p = 0.04), hypertensive (85%, p <0.01), and diabetic (49%, p = 0.04). The cohort with the highest CPK-MB release (CPK-MB >3 times the upper limit of normal) had significantly higher rates of adverse clinical events at 12 months (major adverse cardiac events 40%, p <0.01), including death (9.3%, p <0.01) and late thrombosis (6.3%, p <0.01). Periprocedural CPK-MB elevation is of prognostic importance in patients treated with IRT for ISR, and its analysis appears to be mandatory to risk stratify these patients. The impact of glycoprotein IIb/IIIa antagonists in reducing periprocedural CPK-MB release awaits evaluation. PMID- 14759381 TI - Differential contribution of dead space ventilation and low arterial pCO2 to exercise hyperpnea in patients with chronic heart failure secondary to ischemic or idiopathic dilated cardiomyopathy. AB - In chronic heart failure (CHF), the abnormally large ventilatory response to exercise (VE/VCO(2) slope) has 2 conceptual elements: the requirement of restraining arterial partial pressure of carbon dioxide (pCO(2)) from increasing (because of an increased ratio between increased physiologic dead space and tidal volume [VD/VT]) and the depression of arterial pCO(2) by further increased ventilation, which necessarily implies an important non-carbon dioxide stimulus to ventilation. We aimed to assess the contribution of these 2 factors in determining the elevated VE/VCO(2) slope in CHF. Thirty patients with CHF underwent cardiopulmonary exercise testing (age 65 +/- 11 years, left ventricular ejection fraction 34 +/- 15%, peak oxygen uptake 15.2 +/- 4 ml/kg/min, VE/VCO(2) slope 36.4). At rest and during exercise, arterial pCO(2) was measured and VD was calculated and separated into serial and alveolar components. VD/VT decreased from 0.57 at rest to 0.44 at peak exercise (p <0.01). VE/VCO(2) slope was correlated with peak exercise VD/VT (r = 0.67), the serial VD/VT ratio (r = 0.64), and alveolar VD/VT ratio (r = 0.51) at peak exercise (all p <0.01). VE/VCO(2) slope was also correlated with arterial pCO(2) (r = -0.75, p <0.001). Despite this, arterial pCO(2) was not related to peak oxygen uptake (r = 0.2) or to arterial lactate (r = -0.25) and only weakly to New York Heart Association functional class (F = 3.7). First, the increased VE/VCO(2) slope was caused by both the high VD/VT ratio and by other mechanisms, as shown by low arterial pCO(2) during exercise. Second, this latter component (depression of arterial pCO(2)) was not related to conventional measures of heart failure severity. PMID- 14759382 TI - Usefulness of tissue Doppler imaging for estimation of filling pressures in patients with primary or secondary pure mitral regurgitation. AB - Mitral annular velocities derived from tissue Doppler (S', E', A') and left ventricular (LV) end-diastolic pressures were obtained in 11 patients with significant primary mitral regurgitation (MR), 26 patients with significant MR secondary to ischemic or dilated cardiomyopathy, and in 29 asymptomatic controls. The mitral E/E' ratio was related significantly to LV end-diastolic pressure in patients with secondary, but not in patients with primary MR. In patients with secondary MR, a mitral E/E' ratio >15 predicted an elevated LV end-diastolic pressure with a sensitivity of 80% and a specificity of 100%. Thus, the mitral E/E' ratio is a reliable estimate of filling pressures only in subjects with significant secondary, but not with significant, primary MR. PMID- 14759383 TI - Rate control in patients with chronic atrial fibrillation. PMID- 14759384 TI - Comparison of the predictive value of exercise-induced ST depression versus exercise technetium-99m sestamibi single-photon emission computed tomographic imaging for detection of coronary artery disease in patients with left ventricular hypertrophy. AB - Scant knowledge exists regarding the significance of either additional ST depression in the presence of baseline depression or new stress-induced ST depression in patients with left ventricular (LV) hypertrophy. Accordingly, the purpose of this investigation is to determine whether the appearance and/or severity of ST abnormalities during exercise stress testing can accurately predict the prevalence of ischemic burden as measured by quantitative technetium 99m-sestamibi single-photon emission computed tomographic imaging in patients with LV hypertrophy. The results show that the presence of exercise ST-segment depression, as well as its magnitude, are not accurate predictors for identifying patients with coronary artery disease in the presence of electrocardiographic criteria for LV hypertrophy. PMID- 14759385 TI - Prevalence of anxiety in coronary patients with improvement following cardiac rehabilitation and exercise training. AB - Numerous studies have indicated that psychological distress, including anxiety, is a significant risk factor for coronary artery disease (CAD). We studied 500 consecutive patients after recent CAD events and demonstrated a very high prevalence of generalized anxiety and moderate to severe anxiety symptoms, particularly in younger patients. In addition, our data demonstrate the abnormal CAD risk profiles in these anxious patients with CAD and the marked improvements in the overall risk profiles, levels of anxiety, and overall quality of life after cardiac rehabilitation and exercise training programs. Greater attention at detecting and treating chronic anxiety, particularly with formal cardiac rehabilitation, is needed for the secondary prevention of CAD. PMID- 14759386 TI - Effect of preintervention remodeling type on subsequent coronary artery behavior after directional atherectomy. AB - To evaluate the influence of preintervention remodeling on subsequent vessel behavior after directional coronary atherectomy (DCA) under intravascular ultrasound (IVUS) guidance, serial (before and after DCA and at 6-month follow up) IVUS data were analyzed for 246 lesions that were classified into 2 categories: positive remodeling (PR) in 77 lesions versus intermediate or negative remodeling in 169 lesions. Although the 2 groups had similar baseline characteristics, IVUS data showed that the PR group had a greater acute lumen area (LA) gain without an increased late LA loss, resulting in a greater net (acute plus late) LA gain and follow-up LA. This suggests that IVUS-guided DCA may neutralize the negative impact of preintervention PR on late vessel patency. PMID- 14759387 TI - Effect of caffeine administered intravenously on intracoronary-administered adenosine-induced coronary hemodynamics in patients with coronary artery disease. AB - It is believed that caffeine attenuates the coronary hyperemic response to adenosine by blocking the A2a receptors. We studied the effect of caffeine on adenosine-induced myocardial hyperemia measured by fractional flow reserve after intracoronary adenosine administration. Fractional flow reserve was not affected by intravenous caffeine infusion at a dose that was comparable to oral consumption. PMID- 14759388 TI - Comparison of frequency of hemorrhagic stroke in patients <75 years versus > or =75 years of age among patients receiving glycoprotein IIb/IIIa inhibitors during percutaneous coronary interventions. AB - We identified 1,369 consecutive patients who received glycoprotein IIb/IIIa inhibitors during 1,461 stenting procedures (2,382 lesions); of these, 240 (17.5%) were aged > or =75 years (253 procedures, 430 lesions). Very elderly patients (> or =75 years) had similar in-hospital outcomes but a higher hemorrhagic stroke rate than patients aged <75 years. PMID- 14759389 TI - Relation between late patency of the infarct-related artery, left ventricular function, and clinical outcomes after primary percutaneous intervention for acute myocardial infarction (CADILLAC trial). AB - The importance of sustained patency of the infarct-related artery after primary percutaneous coronary intervention for acute myocardial infarction is controversial. We examined serial measures of left ventricular function and clinical outcomes in 280 patients with an initially occluded infarct artery in whom Thrombolysis In Myocardial Infarction trial grade 3 flow was achieved and routine follow-up angiography was performed 7 months after percutaneous coronary intervention. Reocclusion of the infarct artery was associated with decreased event-free survival, and the degree of restenosis was an independent predictor of the lack in improvement in left ventricular ejection fraction over time. PMID- 14759390 TI - Outcomes in patients with normal serum creatinine and with artificial renal support for acute renal failure developing after coronary artery bypass grafting. AB - This retrospective study of cardiac surgical patients with normal serum creatinine who developed acute renal failure requiring artificial renal support was undertaken to (1) determine the prevalence of acute renal failure and hospital mortality in this subgroup, (2) identify the independent predictors of early mortality, and (3) determine long-term survival and prognosis. PMID- 14759391 TI - Effectiveness of bivalirudin as a replacement for heparin during cardiopulmonary bypass in patients undergoing coronary artery bypass grafting. AB - We investigated the use of bivalirudin as an anticoagulant therapy during cardiopulmonary bypass in 20 patients who underwent coronary artery bypass grafting. Primary end points consisted of clinical outcome data, whereas secondary end points focused on blood loss, transfusions, pharmacokinetics, and monitoring. Our data provide the first evidence of clinical feasibility of anticoagulation with bivalirudin during cardiopulmonary bypass. PMID- 14759392 TI - Duration of residence in the United States as a new risk factor for coronary artery disease (The Konkani Heart Study). AB - A survey conducted in a relatively homogeneous group of 527 Konkani subjects revealed a high prevalence of coronary artery disease (CAD) despite a lack of smoking and significant obesity. Traditional risk factors (age, gender, high cholesterol, hypertension, and diabetes) were significant predictors of CAD. In addition, duration of residence in the United States (US) emerged as a new independent risk factor. Independent predictors of CAD included age, gender, ever smoking, and duration of residence in the US. We conclude that acculturation may be a major risk factor for CAD in immigrant populations. PMID- 14759393 TI - Effects of rosiglitazone on endothelial function, C-reactive protein, and components of the metabolic syndrome in nondiabetic patients with the metabolic syndrome. AB - Fifty nondiabetic patients who met a modified National Cholesterol Education Program definition for the metabolic syndrome were randomized to receive either rosiglitazone (4 mg/day; n = 25) or placebo (n = 25) for 8 weeks. Compared with those receiving placebo, patients in the rosiglitazone group achieved significant reductions in fasting plasma insulin levels (-40%), homeostasis model assessment indexes (-45%), systolic and diastolic blood pressures, and high-sensitivity C reactive protein levels (-31%). There were no changes in fasting plasma glucose with either treatment. Although rosiglitazone treatment greatly increased plasma levels of low-density lipoprotein cholesterol (18%) and apolipoprotein B (16%), it significantly improved both endothelium-dependent flow-mediated vasodilation (p <0.001) and endothelium-independent nitroglycerin-induced vasodilation (p = 0.01) of the right brachial artery. PMID- 14759394 TI - Effect of direct-current cardioversion on ischemia-modified albumin levels in patients with atrial fibrillation. AB - Ischemia-modified albumin (IMA), measured with the albumin cobalt binding test, is a marker of myocardial ischemia. We measured IMA concentrations after elective direct-current cardioversion for atrial fibrillation to determine whether transient myocardial ischemia occurs. Patients with electrocardiographic changes after cardioversion (ST-depression and/or T-wave inversion) had significantly higher IMA levels than those without these changes. Thus, elevated levels of IMA after cardioversion may reflect transient myocardial ischemia. PMID- 14759395 TI - P-wave dispersion for predicting maintenance of sinus rhythm after cardioversion of atrial fibrillation. AB - P-wave measurements and left atrial function were investigated to predict the maintenance of sinus rhythm after cardioversion of atrial fibrillation. Left atrial dimension <45 mm (p = 0.02) and P-wave dispersion <46 ms (p <0.001) were independent predictors of sinus rhythm maintenance, with a sensitivity of 89% and 96%, respectively. Duration of atrial fibrillation, maximum P-wave duration, and no spontaneous echocardiographic contrast were also univariate predictors. PMID- 14759396 TI - Implications of cardiac resynchronization therapy and prophylactic defibrillator implantation among patients eligible for heart transplantation. AB - This study analyzed the relations and time-related changes in eligibility for cardiac resynchronization therapy and prophylactic defibrillator implantation in 161 potential candidates for heart transplantation. Although up to 62% of patients who fulfilled the severity criteria for heart transplantation were eligible for either device, this percentage increased as clinical/instrumental parameters of heart failure severity worsened. PMID- 14759397 TI - Effectiveness of atrial fibrillation surgery in patients with hypertrophic cardiomyopathy. AB - We report the results of atrial fibrillation surgery in 10 patients with hypertrophic cardiomyopathy, which is the largest case series to date. The Maze procedure, with concomitant septal myectomy if indicated, appears to be feasible in patients with hypertrophic cardiomyopathy and refractory atrial fibrillation. PMID- 14759398 TI - Doppler tissue imaging analysis of ventricular function after surgical and transcatheter closure of atrial septal defect. AB - The long-term ventricular function of children after surgical and transcatheter closure of secundum atrial septal defects was assessed by Doppler tissue imaging and compared with that of age-matched controls. Long-term right ventricular function was impaired in patients after surgical repair, as reflected by decreased systolic and diastolic tricuspid annular velocities and an increased Tei index. In contrast, right ventricular function was preserved after transcatheter device closure. PMID- 14759399 TI - Comparison of survival and other complications after heart transplantation in patients taking amiodarone before surgery versus those not taking amiodarone. AB - Concern exists regarding preoperative amiodarone use and complications occurring after heart transplantation (HT). During a 10-year period of performing HT, we found that patients treated with amiodarone before undergoing HT had decreased overall survival, required longer periods of ventilatory support, and were at increased risk for significant bleeding complications. These findings raise further concern about the use of amiodarone treatment in patients with heart failure before undergoing HT. PMID- 14759400 TI - Influence of age, gender, body mass index, and functional capacity on heart rate variability in a cohort of subjects without heart disease. AB - We studied the time- and frequency-domain indexes of heart rate variability (HRV) in 653 patients without any evidence of heart disease relative to age, gender, heart rate, body mass index, and functional capacity. There was an inverse correlation of HRV with heart rate (p <0.001). HRV indexes decreased with increasing age, differed by gender, and were higher in patients with higher functional capacity. No correlation was noted between HRV and body mass index. PMID- 14759401 TI - Effects of cocaine on heart rate variability in healthy subjects. AB - In this study, we found that cocaine decreased the high-frequency power of heart rate variability in normal volunteers, implicating a decrease in vagal tone. However, the results of our autonomic blocking studies indicated the opposite, suggesting a major influence of cardiac sympathetic drive on the high-frequency power of heart rate variability. PMID- 14759402 TI - Thoracoscopy-assisted Heller myotomy for the treatment of achalasia: results of a minimally invasive technique. AB - BACKGROUND: Several surgical methods have been described to treat achalasia with a recent trend toward utilizing minimally invasive techniques to perform a myotomy. Since 1998 our institution has utilized a minimally invasive thoracoscopy-assisted technique (ThAM) that allows a myotomy to be performed under direct visualization. METHODS: From 1992 to 2002, 57 patients underwent transthoracic Heller myotomy at our institution. Thirty-eight patients (67%) who underwent ThAM were reviewed and compared with 19 (33%) who previously underwent myotomy through a standard open left thoracotomy (OM). RESULTS: There were no operative deaths in the ThAM group (n = 38) and 4 patients (11%) experienced minor morbidity. Four ThAM patients required conversion to open thoracotomy and 2 were lost to follow-up. Of the remaining 32 patients, 29 have improved postoperative dysphagia scores after a mean follow-up of 17 months. Only 4 patients have required further endoscopic or surgical intervention. Compared with the OM group, ThAM patients experienced significantly shorter average surgery time (97 versus 139 minutes), less blood loss (80 versus 155 mL), less postoperative narcotic requirement (8 versus 20 days), and shorter recovery to normal activity (20 versus 73 days). CONCLUSIONS: Thoracoscopy-assisted myotomy results in excellent relief of dysphagia in the short term and would be expected to have long-term results similar to OM. Shorter operating and recovery times as compared with OM without the need for an antireflux procedure makes ThAM an attractive minimally invasive technique. PMID- 14759403 TI - Barrett's esophagus: the role of laparoscopic fundoplication. AB - BACKGROUND: To review our early operative results and endoscopic findings after laparoscopic fundoplication for Barrett's esophagus (BE). METHODS: From January 1995 through December 2000, 49 patients with BE (35 men and 14 women) underwent laparoscopic antireflux surgery. Median age was 54 years (range, 28 to 85 years). No patient had high-grade dysplasia; 6, however, had low-grade dysplasia. All 49 patients had gastroesophageal reflux symptoms. Heartburn was present in 41 patients (84%), dysphagia in 16 (33%), epigastric or chest pain in 9 (18%), and other symptoms in 16 (33%). A Nissen fundoplication was performed in 48 patients and a partial posterior fundoplication in 1. Forty-one patients (84%) had concomitant hiatal hernia repair. RESULTS: There were no deaths. Complications occurred in 2 patients (4%). Follow-up was complete in 48 patients (98%) and ranged from 1 to 81 months (median, 29 months). Functional results were classified as excellent in 33 patients (69%), good in 9 (19%), fair in 5 (10%), and poor in 1 (2%). Thirty-three patients (67%) underwent postoperative surveillance esophagoscopy with biopsy. Nine patients (18%) had total regression of BE and 3 (6%) had a decrease in total length. In the 6 patients with preoperative low-grade dysplasia, dysplasia was not found in 4, remained unchanged in 1, and progressed to in situ adenocarcinoma in 1. CONCLUSIONS: Laparoscopic fundoplication is effective in controlling symptoms in the majority of patients with BE. While disappearance of BE may occur in some patients, the possibility of developing esophageal adenocarcinoma is not eliminated by laparoscopic fundoplication. Therefore, endoscopic surveillance should continue. PMID- 14759404 TI - Closure of large intrathoracic airway defects using extrathoracic muscle flaps. AB - BACKGROUND: Prospective assessment of pedicled extrathoracic muscle flaps for the closure of large intrathoracic airway defects after noncircumferential resection in situations where an end-to-end reconstruction seemed risky (defects of > 4-cm length, desmoplastic reactions after previous infection or radiochemotherapy). METHODS: From 1996 to 2001, 13 intrathoracic muscle transpositions (6 latissimus dorsi and 7 serratus anterior muscle flaps) were performed to close defects of the intrathoracic airways after noncircumferential resection for tumor (n = 5), large tracheoesophageal fistula (n = 2), delayed tracheal injury (n = 1) and bronchopleural fistula (n = 5). In 2 patients, the extent of the tracheal defect required reinforcement of the reconstruction by use of a rib segment embedded into the muscle flap followed by temporary tracheal stenting. Patient follow-up was by clinical examination bronchoscopy and biopsy, pulmonary function tests, and dynamic virtual bronchoscopy by computed tomographic (CT) scan during inspiration and expiration. RESULTS: The airway defects ranged from 2 x 1 cm to 8 x 4 cm and involved up to 50% of the airway circumference. They were all successfully closed using muscle flaps with no mortality and all patients were extubated within 24 hours. Bronchoscopy revealed epithelialization of the reconstructions without dehiscence, stenosis, or recurrence of fistulas. The flow volume loop was preserved in all patients and dynamic virtual bronchoscopy revealed no significant difference in the endoluminal cross surface areas of the airway between inspiration and expiration above (45 +/- 21 mm(2)), at the site (76 +/- 23 mm(2)) and below the reconstruction (65 +/- 40 mm(2)). CONCLUSIONS: Intrathoracic airway defects of up to 50% of the circumference may be repaired using extrathoracic muscle flaps when an end-to-end reconstruction is not feasible. PMID- 14759406 TI - Management of postintubation membranous tracheal rupture. AB - BACKGROUND: Postintubation tracheobronchial laceration is a rare complication of general anesthesia. A renewed interest in this disorder induced us to review our experience on its treatment, focusing on the evolution of the surgical approach, and describing a technical variation of the transcervical approach. METHODS: From January 1994 to December 2002 we treated 13 patients with diagnosis of postintubation tracheobronchial laceration. The treatment was nonsurgical in 3 patients (1-cm-long tear) and surgical in the other cases. Two lesions extending to the main bronchi were repaired through a right thoracotomy as well as four lesions limited to the trachea observed before January 2001. After this date we used the transcervical approach for entirely intratracheal lesions: in three cases we performed an anterior transverse tracheotomy and in one case a transverse and midline vertical incision (T tracheotomy). RESULTS: Both conservative and surgical therapy were successful in all the cases. Two patients in the thoracotomy group had a transient right vocal cord palsy. No morbidity was observed with the cervical approach. Normal healing of the sutures was evidenced by an endoscopic follow-up 30 days later. CONCLUSIONS: In our experience nonsurgical treatment is advisable in small (length < 2 cm) uncomplicated tears. Concerning surgery, thoracotomy is indicated in tracheal lacerations extending to the main bronchi, whereas the transcervical approach is preferred for intratracheal tears because of its efficacy in reaching and suturing the lesions extending to the carina and for its limited invasiveness. PMID- 14759407 TI - Thoracoscopic sympathectomy for hyperhidrosis: indications and results. AB - BACKGROUND: Hyperhidrosis can cause significant professional and social handicaps. Although treatments such as oral medication, botox, and iontophoresis are available, surgical sympathectomy is being increasingly utilized. METHODS: Between January 1997 and December 2002, 180 patients with palmar, axillary, facial, or plantar hyperhidrosis underwent a thoracoscopic sympathectomy. Surgical technique evolved during our study period and included excision of the sympathetic ganglia at T(2), T(3), or T(4) depending on the location of the sweating using monopolar cautery. RESULTS: Patient demographics included 33% males (59/180) and 67% females (121/180), with a mean age of 29.2 years old (range 12 to 76 years old). Ethnic origin was 67% white (122/180), 19% Asian (34/180), 8% Black (14/180), and 6% Hispanic (10/180). Positive family history of hyperhidrosis was noted in 57%. Preoperatively, 49% patients (86/180) had palmar sweating only, 7% patients (12/180) axillary only, 24% patients (43/180) palmar and axillary, 16% patients (28/180) face/scalp only, and 7% patients (11/180) all of the above; additionally 69% patients (125/180) had plantar hyperhidrosis. All procedures were performed through 3-mm and 5-mm ports, and 98% (177/180) were completed as an outpatient procedure. Complications included a mild temporary Horner's Syndrome (n = 1; 0.5%), air leak requiring chest drainage (n = 9; 5%), and bleeding (n = 3; 1.6%) requiring thoracoscopic reexploration (n = 1) and chest drainage (n = 2). Success rates were palmar 100% (109/109), axillary 98% (48/49), and face/scalp 93% (26/28). Plantar hyperhidrosis responded with improvement in 82% (72/88) of all patients. Seventy-eight percent patients (96/123) experienced compensatory hyperhidrosis, usually affecting the stomach, chest, back, and neck. Overall satisfaction was 94% (139/148). CONCLUSIONS: Thoracoscopic sympathectomy is a safe and effective outpatient method for managing hyperhidrosis. Although overall satisfaction is high, patients should be fully informed about the potential for compensatory sweating. PMID- 14759408 TI - Optimal distance of malignant negative margin in excision of nonsmall cell lung cancer: a multicenter prospective study. AB - BACKGROUND: Complete excision of nonsmall cell lung cancer is necessary during a limited resection procedure, as a malignant positive margin can lead to margin relapse. Because there is scant information available regarding the optimal size of a malignant negative margin, we conducted a multicenter, prospective study to more fully elucidate this area of concern. METHODS: Two hundred five pulmonary tumors (22 nonsmall cell lung cancers and 183 undiagnosed lesions) were excised, of which 118 nonsmall cell lung cancer lesions were analyzed. Malignant status was considered positive when either a cytologic or histologic technique revealed the margin to be malignant. Maximum tumor diameter (from 4 to 45 mm with an average of 15.3 mm), margin distance (from 0 to 25 mm with an average of 9.3 mm), tumor location, extent of stapling carried out, and performance of a thoracotomy were the variables. RESULTS: Seventy-two of the sample tissues (61%) were malignant negative. The negative group had smaller maximum tumor diameter, greater margin distance, lesions in more easily resectable regions, and more often required stapling only. Using a multivariate analysis, maximum tumor diameter and margin distance were found to be independent factors. The number of malignant negative margins was 7/7 (100%) when the margin distance was greater than 20 mm, and the number of malignant negative margins was 21 of 21 (100%) when the resected tumors had a margin distance greater than the maximum tumor diameter. CONCLUSIONS: Malignant positive margins were not found when the margin distance was greater than the maximum tumor diameter, which was considered to be the optimal margin distance for prevention against margin relapse. PMID- 14759409 TI - Nonanatomic prognostic factors in resected nonsmall cell lung carcinoma: the importance of perineural invasion as a new prognostic marker. AB - BACKGROUND: A number of prognostic factors have been reported for resected nonsmall cell lung carcinoma. Although none of them reported to have greater prognostic impact than the TNM staging system, which is based on anatomical descriptions of tumors, the prognoses of a significant number of patients are not in agreement with real survival of the patients. Moreover, certain histopathologic properties of the tumor (such as lymphatic and vascular invasion) could help to predict the survival of the patients. METHODS: A retrospective study was conducted on 82 surgically resected nonsmall cell lung carcinomas, and the following prognostic factors were evaluated in univariate analysis: age, gender, clinical and surgical-pathologic T and N status, histologic type of tumor, grade of differentiation, lymphatic invasion, vascular invasion, and perineural invasion. RESULTS: Lymphatic invasion and perineural invasion of the tumor were found to be significant prognostic factors (p = 0.02 and p = 0004). Blood vessel invasion (venous or arterial involvement) had no prognostic impact (p > 0.05). According to multivariate analyses, three factors were selected as prognostic indicators: (1) clinical N status (p = 0.027), (2) lymphatic invasion (p = 0.027) and (3) perineural invasion (p = 0.0148). By combining these factors we identified a poor prognostic subgroup of patients with stage I disease. CONCLUSIONS: Our study showed that lymphatic vessel and perineural invasion of the tumor could be prognostic factors, along with anatomical determinants such as cN and surgical-pathologic stage of the pulmonary carcinoma. PMID- 14759410 TI - Radioisotope lymph node mapping in nonsmall cell lung cancer: can it be applicable for sentinel node biopsy? AB - BACKGROUND: Previous studies on intrathoracic lymph node mapping have focused on the validity of a sentinel node concept, but not on the usefulness for sentinel node biopsy. METHODS: The subjects were 15 patients clinically diagnosed with N0 nonsmall cell lung cancer. Technetium-99m tin colloid was injected into the peritumoral area 1 day preoperatively and a time course of tracer migration was monitored by scintigraphy. A hand-held gamma probe counter was used to count the intrathoracic lymph node stations. Resected nodes were also counted to assess the accuracy of the intrathoracic counting. RESULTS: Serial scintigraphies showed that the tracer migrated through airways and the appearance resembled hot nodes. On intrathoracic counting, 50% of the nodal stations appeared positive; however, only 23% of these apparently positive nodal stations were ultimately shown to be truly radioactive. The true positive and true negative rates of detecting intrathoracic hot nodes were 100% and 56%, respectively. Because the counts of the nodal stations could include the counts from the hot primary tumor ("shine through") or airway radioactivity, legitimate hot nodes were identified after dissecting all the apparently positive nodal stations. Two of the 9 patients in whom hot nodes were identified had nodal metastatic disease and actually had tumor cells within the hot nodes. The only complication related to the preoperative injection of technetium-99m was a minor pneumothorax. CONCLUSIONS: Although radioisotope intrathoracic lymph node mapping is safe, it appears to be unsuitable for sentinel node biopsy because shine-through and the airway-migrated radioactive tracer complicated the intrathoracic counting. Only serial scintigraphy could distinguish hot nodes from airway migration. PMID- 14759411 TI - Surgical resection of lung metastases from epithelial tumors. AB - BACKGROUND: One of the main characteristics of malignant tumors is the capability to disseminate, giving rise to local or distant metastases. Pulmonary metastases occur in almost 30% of all oncology patients, and secondary lung tumors are more frequent than primary ones. Surgical resection of metastases is now a well established procedure, but the results vary with the histologic type of the primary tumor. We wished to identify factors affecting late survival after pulmonary metastasectomy for epithelial tumors. METHODS: We have reviewed retrospectively data for 78 patients who underwent 101 curative resections of epithelial pulmonary metastases between January 1988 and December 2000 at our department. Potential prognostic factors affecting late survival, namely histology of the primary tumor, disease-free interval, number and size of resected lung metastases, involvement of lymph nodes, use of nonsurgical adjuvant therapy, and relapse of pulmonary metastases, were investigated. RESULTS: There was no operative mortality, and the postoperative course was uneventful in 91.1% of the procedures. The mean disease-free interval was 48.2 +/- 59.8 months, and the mean follow-up after the first pulmonary metastasectomy was 40.8 +/- 31.5 months. Mean overall survival was 81.0 +/- 10.0 months, and 5-year and 10-year survival rates were 47.4% and 37.7%, respectively. By univariate and multivariate analyses, the disease-free interval and the prognostic grouping system proposed by the International Registry of Lung Metastases were found to significantly influence the long-term survival. Presence of symptoms also showed an important influence. CONCLUSIONS: Resection of epithelial pulmonary metastases is safe and effective, and is associated with very low perioperative morbidity and mortality and a reasonable 5-year and 10-year survival. In the present study, the disease free interval influenced significantly the long-term survival. Our results did not differ significantly from (and in many cases compared favorably with) those described in the literature. PMID- 14759412 TI - How long can we preserve the pulmonary graft inside the nonheart-beating donor? AB - BACKGROUND: The use of lungs from nonheart-beating donors (NHBD) might significantly alleviate the organ shortage. Extending the preharvest interval in NHBD would facilitate distant organ retrieval. We hypothesized that prolonged topical cooling inside NHBD after 60 minutes of initial warm ischemia would not affect the pulmonary graft. METHODS: Domestic pigs were anesthetized and divided into three groups (n = 6 in each group). In the control group (HBD), lungs were flushed, explanted, and further stored in low potassium dextran solution (4 degrees C) for 4 hours. In the two study groups pigs were sacrificed by myocardial fibrillation and left untouched for 1 hour. Chest drains were then inserted for topical lung cooling (6 degrees C) for 3 hours (NHBD-TC3) or 6 hours (NHBD-TC6). The left lung in all groups was then prepared for evaluation. In an isolated circuit lungs were ventilated and reperfused through the pulmonary artery. Hemodynamic, aerodynamic, and oxygenation variables were measured 35 minutes after onset of controlled reperfusion. Wet-to-dry weight ratio was calculated. RESULTS: No significant differences were observed among the three groups in pulmonary vascular resistance (p = 0.38), mean airway pressure (p = 0.39), oxygenation index (p = 0.62), and wet-to-dry weight ratio (p = 0.09). CONCLUSIONS: These data confirm that 1 hour of warm ischemia does not affect the pulmonary graft from NHBD compared with HBD. The preharvest interval can be safely extended up to 7 hours postmortem by additional topical cooling of the graft inside the cadaver. This technique may facilitate distant organ retrieval in NHBD. PMID- 14759413 TI - Lateral thoracic expansion for Jeune syndrome: evidence of rib healing and new bone formation. AB - BACKGROUND: Lateral thoracic expansion is a procedure that has been described to enlarge the thoracic cage in patients with Jeune's asphyxiating thoracic dystrophy. The procedure involves separating ribs from their periosteum and plating them together in an expanded fashion with titanium struts. We have speculated that the ribs heal in this situation, despite the absence of surrounding periosteum, and that new rib formation occurs in the liberated periosteum. METHODS: Radiographic studies of patients who have undergone lateral thoracic expansion were reviewed for evidence of rib healing or periosteal new bone formation. RESULTS: This study presents radiologic evidence that rib healing actually occurs, as does periosteal ossification. CONCLUSIONS: Lateral thoracic expansion creates additional chest wall that is formed of autologous tissue, fully healed, and not ultimately dependent on titanium struts. PMID- 14759414 TI - A new animal model for pulmonary hypertension based on the overexpression of a single gene, angiopoietin-1. AB - BACKGROUND: Angiopoietin-1 gene expression in human pulmonary hypertensive lungs is directly proportional to increasing pulmonary vascular resistance. We hypothesized that targeted overexpresssion of angiopoietin-1 in the lung would cause persistent pulmonary hypertension in an animal model. METHODS: We injected 2 x 10(10) genomic particles of adeno-associated virus-angiopoietin-1 (AAV-Ang-1) into the right ventricular outflow tract of 30 Fischer rats while using adeno associated virus-lacZ (AAV-lacZ) injected rats and carrier-injected rats as our control groups. All animals underwent survival surgery and were sacrificed at serial timepoints postgene delivery. At each timepoint, pulmonary artery pressures were measured and pulmonary angiography using the Microfil polymer perfusion technique was performed. The lungs were harvested for pathologic analysis, mRNA analysis, Western blot assays, and in situ RNA hybridization to localize gene expression. RESULTS: Pulmonary artery pressures of AAV-Ang-1 injected rats were significantly increased compared with the control groups (p < 0.01) at all timepoints. Pathologic analysis of AAV-Ang-1 lung specimens demonstrated increased smooth muscle cell proliferation within the medial layer of arterioles with obliteration of small vessels similar to that seen in human pulmonary hypertension. Angiograms of AAV-Ang-1 injected lungs showed blunting of small peripheral arterioles consistent with advanced pulmonary hypertension. In situ RNA hybridization localized angiopoietin-1 expression to the vascular wall of small-caliber pulmonary vessels. Protein and mRNA assays confirmed persistent angiopoietin-1 expression in the lung for up to 60 days postgene delivery. CONCLUSIONS: Overexpression of angiopoietin-1 using an adeno-associated virus vector causes pulmonary hypertension in rats. These data provide a novel physiologic animal model for pulmonary hypertension. PMID- 14759416 TI - Induction of pulmonary angiogenesis by adenoviral-mediated gene transfer of vascular endothelial growth factor. AB - BACKGROUND: We hypothesized that gene transfer of vascular endothelial growth factor (VEGF) mediated by an adenovirus vector might induce pulmonary artery angiogenesis in a lamb model of pulmonary artery hypoplasia. METHODS: Thirteen fetal lambs had left pulmonary artery banding at 106 days of gestation. Following birth, 3 groups were divided: VEGF group (n = 5) and beta-GAL group (n = 4) received an adenoviral vector encoding respectively for human VEGF165 and for galactosidase A. A control group (n = 4) had neither gene nor virus. Viral suspensions were selectively instilled in the left bronchus 6.5 days after birth. Five nonoperated lambs constituted the normal group. Euthanasia was performed at 30 days of age. Gene transfer was confirmed by blue coloration of left lung obtained with Xgal solution in an additional experiment. Histomorphometric evaluation was performed. All groups were compared with ANOVA test and paired test was used to compare right and left lung in each animal. RESULTS: Left lung was similarly hypoplastic in all operated lambs. Left pulmonary artery hypoplasia present in all operated groups was significantly less pronounced in VEGF group. The number of pleural arteries was similarly increased in left lung of all operated lambs. Left lung arterial density was higher in VEGF group than in all other groups. The percentage of parenchyma of left lung was lower in beta-GAL group than in all others, partially returned to normal in VEGF group. CONCLUSIONS: In this model, transbronchial VEGF gene transfer induces pulmonary angiogenesis, proximal pulmonary artery growth and contributes to lung parenchyma recovery. PMID- 14759418 TI - Late results of palliative atrial switch for transposition, ventricular septal defect, and pulmonary vascular obstructive disease. AB - BACKGROUND: Palliative atrial switch (PAS) procedures that reroute pulmonary and systemic venous drainage and leave a ventricular septal defect (VSD) open have been used in the treatment of deeply cyanotic patients who have severe pulmonary vascular obstructive disease (PVOD). Palliative atrial switch is beneficial for patients with transposition of the great arteries or other complex lesions with VSD who show higher arterial oxygen saturation in the pulmonary artery than in the aorta (transposition hemodynamics/unfavorable streaming). We reviewed the early and late results of PAS (Mustard, n = 25; Senning, n = 3) in patients at two institutions. METHODS: Between April 1965 and March 2000, PAS was performed in 28 cyanotic patients (18 male, 10 female). Median age was 10 years (range, 1 to 27). Mean preoperative pulmonary arterial pressure was 68 mm Hg (range, 30 to 121 mm Hg). Mean systemic arterial oxygen saturation was 65% (range, 47% to 80%). The majority of patients (95%) were in New York Heart Association (NYHA) functional class III or IV preoperatively. RESULTS: Overall early mortality was 21%; for patients after 1972 (n = 23), the early mortality was 8.7%. Mean follow up was 8.3 years (maximum 20). Mean postoperative systemic arterial oxygen saturation was increased significantly to 88% (p < 0.0001). Late survival for early survivors at 5, 10, and 15 years respectively was 84% (59%, 97%), 64% (39%, 88%), and 54% (15%, 72%). The NYHA functional class was significantly improved; 94% of late survivors (n = 17) were in functional class I or II (p = 0.002). CONCLUSIONS: The PAS operation significantly improves systemic arterial oxygen saturation and quality of life in selected patients with transposition hemodynamics, VSD, and severe PVOD. PMID- 14759419 TI - A new procedure for Ebstein's anomaly. AB - BACKGROUND: A new procedure for correction of Ebstein's anomaly that restores to near normal the anatomic and physiologic function of the tricuspid valve and the right ventricle is reported. METHODS: Between December 1997 and September 2002, 34 consecutive patients with Ebstein's anomaly underwent this new procedure. There were 13 male and 21 female patients aged 9 months to 48 years (mean, 17 years). Tricuspid incompetence was moderate in 12 patients and severe in 22. Our repair technique is as follows: the displaced posterior leaflet with some chordae tendineae and corresponding papillary muscle are detached from the annulus and ventricular wall, respectively. The leaflet is then reattached to the native posterior annulus with reimplantation of the papillary muscle. The displaced septal leaflet is treated in the same manner. Most of the atrialized portion of the ventricular wall is excised; the tricuspid annulus is plicated. In 8 of the patients the septal leaflet was severely hypoplastic and necessitated creation of a new leaflet using autologous pericardium. RESULTS: All patients survived and recovered uneventfully. Postoperative echocardiography showed that tricuspid incompetence disappeared in 29 patients and was mild in 5. Right ventricular size decreased significantly with complete disappearance of the atrialized segment. Follow-up of patients ranged from 1 to 55 months (mean, 25 months), with 9 patients having more than 3 years of follow-up. They are doing well and their exercise tolerance improved to normal. CONCLUSIONS: This new procedure anatomically corrects Ebstein's anomaly with the satisfactory early and midterm results. PMID- 14759421 TI - Effect of a selection and postoperative care protocol on survival of infants with hypoplastic left heart syndrome. AB - BACKGROUND: We report the development and implementation of a program designed to assign patients preoperatively to either transplant or Norwood procedure based on a score derived from known risk factors and to enhance postoperative care of infants undergoing the Norwood procedure. METHODS: A weighted score for each of six variables comprised the scoring system: ventricular function, tricuspid regurgitation, ascending aortic diameter, atrial septal defect blood flow characteristics, blood type, and age. The scoring system was used to prospectively assign mortality risk and lead to recommendation of either Norwood procedure or transplantation. RESULTS: Survival following the Norwood procedure significantly improved after the management program was implemented (88% versus 40% at 48 hours, 57% versus 10% at 30 days, and 50% versus 10% at 1 year, p < 0.0001 at each time point). The survival of the group that received a score of 7 or less (high risk) who underwent the Norwood procedure was 78% at 48 hours, 44% at 30 days, and 33% at 1 year; survival rates among patients considered lower risk (greater than 7) were 100% at 48 hours and 80% at 30 days and 1 year. Transplant outcomes remained unchanged. CONCLUSIONS: We report improved survival following the Norwood procedure after the implementation of an institutional management approach aimed at improving the outcome of infants with hypoplastic left heart syndrome and may help neutralize historical biases toward Norwood procedure or transplantation. PMID- 14759423 TI - Surgical management of aortopulmonary window and associated lesions. AB - BACKGROUND: Aortopulmonary window is a rare congenital heart defect commonly associated with other cardiac anomalies. Although single institutional experience is low, several surgical techniques have been reported. The purpose of this study is to describe our approach to the management of aortopulmonary window and its associated lesions. METHODS: Between February 1996 and November 2002, 13 patients underwent repair of aortopulmonary window. The age range went from 4 days to 5.5 months (mean 42 +/- 52 days), with 9 patients younger than 1 month old. The weight range was from 1.9 to 6.7 kg (mean 3.5 +/- 1.2 kg). Concomitant cardiac anomalies were present in 11 patients. The major additional anomalies were interruption of aortic arch in 4 patients and tracheal stenosis in 1 patient. Initial diagnoses were made using two-dimensional echocardiography only. RESULTS: There was one postoperative death. In general, patients with aortopulmonary window and additional major defects had a prolonged intensive care unit and hospital stay when compared with the other patients. Follow-up time ranged from 2 months to 6.8 years (mean of 2.5 +/- 2.2 years). There were no reoperations and no late deaths. Transcatheter balloon dilatation of the repaired aortic arch was required in 1 patient and of the right pulmonary artery in another. All other patients had good flow to both pulmonary arteries. No residual shunts were detected at the aortopulmonary window site, and pulmonary pressures were normal. CONCLUSIONS: Aortopulmonary window may be effectively diagnosed with echocardiography. Early surgical treatment (neonatal period, if possible) is safe and associated with the best long-term results, even in the presence of other cardiac anomalies. Complete separation and reconstruction of both aorta and pulmonary arteries under direct vision may prevent recurrence and distortion of adjacent structures. PMID- 14759424 TI - Opposite trends in coronary artery and valve surgery in a large multisurgeon practice, 1979-1999. AB - BACKGROUND: Trends in coronary artery bypass (CAB) and valve operations (VO) may help predict the future of cardiac surgery in the context of changing case mix, shifting paradigms, emerging technology, and population demographics. METHODS: We retrospectively reviewed all 30,319 adult CAB and VO in our group from 1979 to 1999 according to specific procedures. RESULTS: Coronary artery bypass volumes peaked in 1996 at 1,895 cases, declining 15.3% to 1,605 cases in 1999 with a decrease in risk profile and percent reoperations and an increase in mean age and percent octogenarians, prior percutaneous coronary interventions (PCI), left internal mammary artery (LIMA) graft usage, off-pump technology usage, and hospital mortality of reoperations. Right internal mammary grafts were employed infrequently and radial artery grafts transiently. Overall VO volumes continued to increase 24.0% since 1996, from 470 to 583 cases with a decreased risk profile, increased mean age, and percent octogenarians and prior PCI. The percentage of mechanical valve implants decreased, while the percentage of various tissue solutions for valve disease increased. Limited access incisions and port-access were employed transiently with CAB and VO. CONCLUSIONS: Coronary artery bypass volumes are decreasing, with an increasing percentage of LIMA grafts and off-pump cases. Valve operation volumes are steadily increasing, with a decreasing percentage of mechanical valve implants, in favor of various tissue solutions. PMID- 14759425 TI - Methylene blue reduces mortality and morbidity in vasoplegic patients after cardiac surgery. AB - BACKGROUND: The discovery of nitric oxide as mediator in cardiac postoperative vasoplegia encourages the use of inhibitory drugs such as methylene blue. This drug has been used with favorable results in isolated cases. The purpose of this article is to analyze the incidence of the postoperative vasoplegic syndrome, to consider its prognosis, and to evaluate the effect of intravenous methylene blue on mortality. METHODS: Cardiac surgery patients were consecutively included. Vasoplegic syndrome was defined by the presence of the following five criteria: (1) hypotension, (2) low filling pressures, (3) high or normal cardiac index, (4) low peripheral resistance, and (5) vasopressor requirements. Those with vasoplegia were randomized to receive 1.5 mg/Kg of methylene blue or a placebo. A p value less than 0.05 was considered significant. RESULTS: Six hundred thirty eight cardiac surgery patients were consecutively included in this study. Fifty six of these patients fulfilled vasoplegia criteria (8.8%) resulting in higher mortality (10.7% or 6 of 56 patients vs 3.6% or 21 of 582 patients; p value = 0.02). Those treated with methylene blue showed morbidity and mortality reductions (0% versus 21.4% or 6 of 28 patients; p value = 0.01). The duration of the vasoplegic syndrome was shorter in those patients treated with the drug, lasting less than 6 hours in all patients. Patients in the control group showed a slower recovery, lasting more than 48 hours in 8 patients (p value = 0.0007). CONCLUSIONS: Vasoplegic postoperative syndrome was seen in 8.8% of all patients. Outcome in patients with vasoplegia was worse with increased morbidity and mortality. The use of methylene blue reduced the high mortality in this population. PMID- 14759426 TI - Clinical outcome after cardiac operations in patients with cirrhosis. AB - BACKGROUND: To evaluate the clinical outcome after cardiac operations in patients with cirrhosis, a retrospective study was undertaken. METHODS: Between 1989 and 2003, 18 patients with cirrhosis who underwent cardiac operations were identified. Their preoperative status and postoperative clinical results were assessed. RESULTS: Ten patients were classified as having Child-Pugh class A cirrhosis, 7 as having class B cirrhosis, and 1 as having class C cirrhosis. Fifteen of 18 patients underwent cardiac surgery using cardiopulmonary bypass, and the remaining 3 patients with class B cirrhosis received coronary artery bypass grafting without cardiopulmonary bypass. In patients undergoing cardiopulmonary bypass, 60% of those with class A cirrhosis and 100% of those with class B cirrhosis and class C cirrhosis had postoperative major complications, including infection, respiratory failure, renal failure, bleeding, and gastrointestinal disorder. One of 3 patients (33%) with class B cirrhosis undergoing coronary artery bypass grafting without cardiopulmonary bypass had major complications. The overall postoperative mortality rate was 17%. Hospital mortality of patients with class A cirrhosis, class B cirrhosis, and class C cirrhosis undergoing cardiopulmonary bypass was 0%, 50%, and 100%, respectively. None of 3 patients with class B cirrhosis undergoing coronary artery bypass grafting without cardiopulmonary bypass died in this study. CONCLUSIONS: Although the incidence of major complications was high, patients with Child-Pugh class A cirrhosis tolerated cardiac surgery satisfactorily. Patients with more advanced cirrhosis, however, may not be suitable for elective cardiac operations with cardiopulmonary bypass. Although our results are not conclusive, coronary artery bypass grafting without cardiopulmonary bypass can be an alternative therapeutic strategy for patients with advanced cirrhosis requiring surgical revascularization. PMID- 14759427 TI - Application of biological glue in repair of intracardiac structural defects. AB - BACKGROUND: BioGlue (Cryolife Inc, Kennesaw GA) was introduced as an alternative tissue sealant. Its most common application has been in repairs of acute dissections of the aorta. There is no reported experience with its use in the repair of intracardiac structural defects. METHODS: In 5 patients BioGlue was used as an adjunct in repairs of complex intracardiac structural defects. It was used during patch repair of posterior mitral annular defects in 2 patients and aortic annular defect in 1 patient in the presence of active endocarditis. It was also used in 1 patient with a chronic atrioventricular groove pseudoaneurysm following mitral valve replacement, and in 1 patient during repair of a postinfarction posterior ventricular septal rupture. RESULTS: There were no hospital or late deaths. Immediate intraoperative transesophageal echocardiography and late follow-up echocardiography documented complete and durable repair of all defects without recurrence. At follow-up all patients are in New York Heart Association class I-II, 6 to 29 months postoperatively. No patient has suffered late complications or exhibited signs of glue embolization. CONCLUSIONS: BioGlue was found to be an effective adjuvant to the standard techniques used for the repair of intracardiac structural defects of various etiologies. Long-term follow-up is recommended to determine its long-term safety in this application. PMID- 14759428 TI - Long-term results of irrigated radiofrequency modified maze procedure in 200 patients with concomitant cardiac surgery: six years experience. AB - BACKGROUND: The Cox maze procedure is considered an effective surgical treatment of atrial fibrillation in patients with and without organic heart disease. Radiofrequency energy offers an alternative to the complex surgical maze procedure. We used the radiofrequency modified maze III procedure in patients with atrial fibrillation undergoing elective concomitant cardiac surgery. This study evaluated the long-term results of the irrigated radiofrequency ablation to create linear lines of conduction block endocardially. METHODS: Between November 1995 and June 2001, 200 patients with mainly structural heart disease and chronic atrial fibrillation underwent intraoperative radiofrequency linear ablation in both atria with concomitant cardiac surgery. RESULTS: The in-hospital mortality rate was 3.5% (7 patients) and during the mean follow-up of 40 months (range, 12 to 80) 27 patients (13.5%) died. Eight patients (4%) were lost from follow-up and complete data were available in 158 survivors. Sinus or atrial rhythm was present in 116 patients (73.4%) and an atrial driven rhythm in 10 patients (6.3%) with an atrioventricular pacemaker. Atrial fibrillation or flutter was documented in 32 patients (20.3%). Antiarrhythmic drugs were used in 49% of survivors who were free of atrial fibrillation or flutter. CONCLUSIONS: Intraoperative radiofrequency endocardial ablation is an effective technique to eliminate atrial fibrillation with promising long-term results. PMID- 14759429 TI - Semirigid partial annuloplasty band allows dynamic mitral annular motion and minimizes valvular gradients: an echocardiographic study. AB - BACKGROUND: Traditional mitral annuloplasty devices include both rigid rings, which restrict annular motion, and soft rings and bands, which can locally deform. Conflicting data exist regarding their impact on annular dynamics. We studied mitral annuloplasty with a semirigid partial band and with a nearly complete rigid ring. METHODS: Intraoperative three-dimensional transesophageal echocardiograms (n = 14) and predischarge transthoracic echocardiograms were retrospectively analyzed in patients undergoing mitral valve repair for degenerative disease with either a rigid ring (n = 77) or a semirigid partial band (n = 38). Each transesophageal echocardiogram was analyzed with TomTec three dimensional software to produce cardiac cycle frame planimetry and to measure device geometry. Actual device sizes provided reference dimensions. Blinded analysis of Doppler data from transthoracic echocardiograms was performed. RESULTS: Validation of the quantitative transesophageal echocardiogram methodology revealed a 1.3% +/- 0.3% (mean +/- standard error of the mean) underestimation of actual linear dimension. With the semirigid partial band, systolic valve orifice area and intertrigonal distance decreased from 6.14 +/- 0.37 to 5.55 +/- 0.24 cm(2) (-9.6%; p = 0.01) and from 2.69 +/- 0.08 to 2.55 +/- 0.13 cm (-5.2%; p = 0.03), respectively. Systolic anterior-posterior distance decreased from 2.1 +/- 0.10 to 1.95 +/- 0.06 cm (-7.1%; p = 0.01) compared with diastole. In contrast, rigid ring orifice area was unchanged (4.12 +/- 0.15 to 4.10 +/- 0.16 cm(2); -0.5%; p = 0.48) during the cardiac cycle. Transthoracic echocardiography revealed significantly lower mitral inflow gradients with semirigid partial band (mean gradients compared with rigid ring, 4.0 +/- 0.3 versus 5.0 +/- 0.3 mm Hg; p = 0.02; peak gradients, 8.9 +/- 0.5 versus 11.1 +/- 0.5 mm Hg; p = 0.01). CONCLUSIONS: Three-dimensional transesophageal echocardiographic measurements of annular dynamics are valid and reliable when discrete annuloplasty devices are present. In contrast to the rigid ring, the semirigid partial band permits more physiologic geometric changes and is associated with lower postoperative mitral valve gradients. PMID- 14759431 TI - Usefulness of a multidetector-row computed tomography scanner for detecting pannus formation. AB - BACKGROUND: Prosthetic valve dysfunction (PVD) as a result of pannus or thrombus formation is an infrequent but serious complication. Currently available diagnostic tools, however, are insufficient to detect a minute pannus and thrombi. The use of a more advanced diagnostic image, multidetector-row computed tomography scanner, may enable us to determine the anatomic and functional causes of PVD. METHODS: Patients who underwent aortic valve replacement with a St. Jude Medical valve were examined by transthoracic echocardiography and cineradiography to diagnose PVD. Sixteen patients with PVD (PVD group) and 12 patients with normal prosthetic valve function (control group) were studied using the multidetector-row computed tomography scanner. The multidetector-row computed tomography findings in 2 patients with PVD were validated by the observations during reoperation. RESULTS: In 13 of 16 patients (81%) in the PVD group and 3 of 12 patients (25%) in the control group, multidetector-row computed tomography demonstrated that an abnormal small tissue, regarded as pannus, was found to extend from the left ventricular septum into the pivot guard. These findings were confirmed by the observations during reoperation in 2 patients in the PVD group. CONCLUSIONS: Multidetector-row computed tomography can be a useful diagnostic technique for anatomic and functional evaluation of PVD as a result of pannus formation. PMID- 14759432 TI - Midterm evaluation of the Sorin Bicarbon heart valve prosthesis: single-center experience. AB - BACKGROUND: The purpose of this study was to perform midterm evaluation of the clinical performance of the Sorin Bicarbon mechanical heart valve prosthesis. METHODS: From November 1992 to December 2002, 328 patients underwent isolated aortic (AVR; 156) or mitral (MVR; 172) valve replacement with the Sorin Bicarbon mechanical valve. Concomitant surgery was performed in 83 patients (25.2%). RESULTS: Total hospital mortality was 5.2%. Survival at 7 years was 79.5% for AVR and 82.4% for MVR. Kaplan-Meier freedoms from valve-related complications were as follows: thromboembolism 92.7% (AVR 94.8%, MVR 92.1%); bleeding 93% (AVR 91.9%, MVR 94.5%); nonstructural dysfunction 96.6% (AVR 94.7%; MVR 97.9%); endocarditis 97.7% (AVR 97.4%, MVR 98.1%); and reoperation 95.7% (AVR 96.6%, MVR 93.9%). Overall freedom from valve-related death was 93.2% (AVR 99.3%, MVR 91.2%). At the end of follow-up, 88.9% of survivors were in New York Heart Association class I or II. CONCLUSIONS: The Sorin Bicarbon valve is a satisfactory mechanical valve prosthesis with low mortality and morbidity and good functional results. PMID- 14759433 TI - Fibrillin and other matrix proteins in mitral valve prolapse syndrome. AB - BACKGROUND: Unlike myxomatous degeneration in Marfan syndrome, which has been reported to result from a mutation in the gene that codes for the extracellular structural protein fibrillin, no specific molecular abnormality has been documented to be the underlying cause of myxomatous degeneration in mitral valve prolapse syndrome (MVPS). The present study examined the distribution of fibrillin and other extracellular matrix proteins in patients with isolated MVPS. METHODS: Mitral valve leaflets from 7 MVPS patients and 5 rheumatic heart disease (RHD) patients were characterized immunohistochemically for fibrillin, elastin, collagen I, and collagen III distribution, and compared with five normal mitral valves. RESULTS: In normal mitral valve leaflets immunostaining for fibrillin, elastin, collagen I, and collagen III revealed a fibrillary and laminar pattern in the atrialis and the spongiosa. In addition, both the collagens were present in the ventricularis, and the coarse bundles in the fibrosa exhibited alternating bandlike collagen I immunoreactivity. The staining patterns of fibrillin, elastin, and collagens I and III revealed distinctly different distribution in MVPS relative to the normal and RHD leaflets. MVPS leaflets in areas of myxoid degeneration displayed a more diffuse, weaker, and nonlaminar pattern of staining for fibrillin. Similar, but less severe abnormality of elastin, collagen I, and collagen III was also observed. Unlike diffuse abnormality in MVPS, the disruption of extracellular proteins in RHD only occurred at the site of the inflammatory damage, but the overall architecture was preserved. CONCLUSIONS: The results of the current study suggest a primary role for abnormal fibrillin and other matrix proteins in producing myxoid degeneration of mitral valve leaflets in MVPS. PMID- 14759434 TI - Role of oral bacterial flora in calcific aortic stenosis: an animal model. AB - BACKGROUND: Calcific aortic stenosis is a major public health problem in the United States. The mechanism of calcification remains unclear. The hypothesis that low grade chronic or recurrent bacterial endocarditis with specific calcifiable bacteria is a cause of calcification of the aortic valves was investigated using an animal model. Such bacteria are typically present as part of the normal human oral flora. METHODS: Forty New Zealand white rabbits were divided into four groups: group 1, control (1 ml of normal saline); group 2, Corynebacterium matruchotti 100,000 colonies; group 3, Streptococcus sanguis II 10 colonies; and group 4, C matruchotti 100,000 colonies plus S sanguis II 10 colonies. Animals were inoculated with bacteria through a flexible catheter placed through the aortic valve through a right carotid cut down. Inoculations were repeated every 3 days the first 2 weeks and then twice a week thereafter. At postmortem examination the aortic valves were harvested, embedded in paraffin, and stained with von Kossa stain. They were also examined by scanning and transmission electron micrography. RESULTS: Group 4 had 93.3% large calcifications (confluent calcium densities that are easily recognized with minimal magnification) and 6.6% small microcalcifications (dustlike microscopic particles requiring a compound microscope to appreciate) of the aortic valves. Group 3 exhibited large calcification in 20% and small in 40% of the aortic valves. Group 1 and group 2 had no evidence of calcification. CONCLUSIONS: These results suggest that recurrent low-grade endocarditis from calcifying oral bacteria, particularly when occurring with synergistic strains, may be one cause of calcific aortic stenosis. PMID- 14759435 TI - The effect of regional ischemia on mitral valve annular saddle shape. AB - BACKGROUND: The mitral valve annulus has a distinctive saddle shape. Recent finite element analysis indicates this shape may contribute to normal valve function by increasing leaflet curvature and reducing leaflet stress. This study tests the hypothesis that acute ischemic mitral regurgitation (AIMR) is associated with loss of annular saddle shape. METHODS: Sonomicrometry array localization (SAL) measured the three-dimensional geometry of the mitral annulus in 6 sheep before and after 30 min of posterior ischemia that produced severe AIMR. Using this SAL data the annular height to commissural width ratio (AHCWR), a measure of annular saddle shape, was calculated throughout the cardiac cycle and reported as a percentage. RESULTS: The normal mitral annulus accentuated its saddle shape rapidly during isovolemic contraction: AHCWR increased from 11.6% +/ 1.1%-13.9% +/- 1.6% (p < 0.001). During ejection AHCWR remained relatively constant ranging from a minimum of 14.1% +/- 1.5% to a maximum of 14.9% +/- 1.3%. During ischemia AHCWR was found to be significantly smaller (p < 0.05) during isovolemic contraction, ejection, and isovolemic relaxation, but not during diastolic filling. Whereas ischemia did not affect AHCWR at end diastole (11.6% +/- 2.8%), the isovolemic accentuation of the saddle shape was lost. CONCLUSIONS: The normal mitral annulus accentuates its saddle shape during systole. This accentuation is eliminated during ischemia that causes AIMR. These data suggest an association between annular saddle shape and valve competency. PMID- 14759436 TI - Comparison of short-term mortality risk factors for valve replacement versus coronary artery bypass graft surgery. AB - BACKGROUND: Risk factors for 30-day operative (short-term) mortality following coronary artery bypass graft (CABG only) procedures are well established. However, little is known about how the risk factors for short-term mortality following valve replacement procedures (with or without a CABG procedure performed) compare with CABG only risk factors. METHODS: Department of Veterans Affairs (VA) records (65,585 records) were collected from October 1991 through March 2001 and analyzed. Risk factors for short-term mortality were compared across three subgroups of patients: CABG only surgery (n = 56,318), aortic valve replacement (AVR) with or without CABG (n = 7450), and mitral valve replacement (MVR) with or without CABG (n = 1817). Multivariable logistic regression analyses were used to compare the relative magnitude of risk for 19 candidate predictor variables across subgroups. RESULTS: Only three patient baseline characteristics differed significantly in magnitude of risk between the procedure groups. Partially or totally dependent functional status significantly increased the risk of short-term mortality for AVR patients (odds ratio [OR] 1.64, 95% confidence interval [CI] 1.29-2.09) and MVR patients (OR 2.21, 95% CI 1.48-3.30), but not for CABG only patients (OR 1.04, 95% CI 0.93-1.16). Conversely, previous heart surgery and New York Heart Association functional class III or IV symptoms conferred greater magnitude of risk for CABG only patients compared with the valve subgroups. CONCLUSIONS: Overall, the risk factors for short-term mortality following valve replacement and CABG surgery appear to be relatively consistent. However, clinicians should be aware of the importance of preoperative functional status as a unique predictor of mortality following valve surgery. PMID- 14759437 TI - Health status and social risk correlates of extended length of stay following coronary artery bypass surgery. AB - BACKGROUND: Preoperative severity of illness in patients undergoing coronary artery bypass grafting (CABG) surgery is a major determinant of clinical postoperative outcomes and surgical length of stay (SLOS). Preoperative patient reported health status and social risk have not been quantified as predictors of SLOS post-CABG. Our hypothesis was that poorer self-reported health and greater social risk, as measured by standardized instruments, are significantly associated with extended SLOS defined as greater than or equal to 7 days. METHODS: In the pilot phase of the Washington State Clinical Outcomes Assessment Program (COAP) patients in a case series between 1995 and 1996 at all hospitals with a cardiac surgery program were administered preoperative SF-36 and Seattle Angina Questionnaires (SAQ) in addition to the collection of prospective clinical data with Society of Thoracic Surgeons' compatible definitions (n = 1073). Factors found significant from bivariate analysis were incorporated into a logistic regression model to assess relative association with extended SLOS (>/= 7 days). RESULTS: The final model included the following elements in descending order of significance: site, SF-36 health perceptions (HP) scale, social risk factors, age, intraaortic balloon pump, congestive heart failure, comorbidity score more than 2, preoperative days more than 2, emergency operation, prior CABG, and gender. CONCLUSIONS: The HP subscore of the SF-36 and the composite social risk factors score were significantly associated with extended SLOS after controlling for other standard clinical variables. "Hospital site" remained the factor with the greatest variance independent of patient severity of illness. PMID- 14759438 TI - Initial experience with sutureless proximal anastomoses performed with a mechanical connector leading to clampless off-pump coronary artery bypass surgery. AB - BACKGROUND: We report our early experience with the Symmetry Aortic Connector (St. Jude Medical, St. Paul, MN) used for sutureless proximal aortosaphenous vein graft anastomoses without any cross clamp during coronary bypass procedures. METHODS: Between November 2001 and August 2002, 206 saphenous vein to aorta proximal anastomoses were created in 132 patients using the Symmetry device. All procedures were performed as part of off-pump coronary artery bypass surgery without any aortic clamping. Intraoperative variables and postoperative data were collected and analyzed retrospectively. RESULTS: All 206 anastomoses (100%) were successfully completed with the connector. Severe atherosclerotic disease of the aorta was documented in 16 patients (12%). Four anastomoses (2%) required additional suture placement. Predeployment problems occurred with 3 grafts (2.5%) during loading of the connector. Average number of distal bypasses was 3.2 per patient. One patient (0.7%) required reoperation for bleeding from a proximal anastomosis. Six patients (4.5%) had perioperative myocardial infarction documented by electrocardiographic changes. Thirty-day operative mortality was 3% (4 patients). Intraoperative transit time flow measurement was performed in all cases (100%). Postoperative angiography in 43 patients at a median 3 months postoperatively revealed occlusion of 9 of the 81 saphenous vein grafts (11%). CONCLUSIONS: The initial experience with a proximal saphenous vein graft to aorta anastomosis using the Symmetry connector demonstrates safety and ease of use. There is however some concern with early graft closure. A prospective randomized study is needed to clarify these concerns. PMID- 14759439 TI - Off-pump versus on-pump myocardial revascularization in low-risk patients with one or two vessel disease: perioperative results in a multicenter randomized controlled trial. AB - BACKGROUND: To evaluate hospital mortality and morbidity after myocardial revascularization in a prospective and multicenter study, comparing on-pump versus off-pump in a special subset of patients with lesions in the left descending artery, alone or associated with the right coronary artery. METHODS: A multicenter prospective randomized study was performed. One hundred and sixty selected low-risk patients were enrolled; 80 patients were operated on-pump (coronary artery bypass grafting [CABG], group I) and 80 patients were operated off-pump (off-pump coronary artery bypass [OPCAB], group II). One hundred and five were male and ages ranged from 39 to 70 years old; mean 58.81 +/- 9.31 and median 59. Preoperative clinical characteristics were similar in both groups; only previous myocardial infarction was higher in the OPCAB group. Patients with severe left ventricular dysfunction (FE 5) coronary artery bypass patients were consecutively treated at our institution. On the basis of the coronary anatomy and possibility of achieving a complete revascularization, 197 patients were assigned to off-pump and 109 to on-pump operation. Overall mortality was 6.2% (19 of 306 patients). RESULTS: Although patients treated off-pump had a better cardiac status, no clinical advantages related to the avoidance of cardiopulmonary bypass were found in the overall population. Off-pump patients had more early and late cardiac complications, whereas patients operated on-pump exhibited an higher incidence of postoperative systemic organ dysfunction. Off-pump surgery improved in-hospital outcome only in the subset of patients at highest risk. CONCLUSIONS: Avoidance of cardiopulmonary bypass does not confer significant clinical advantages in all high risk coronary patients; instead, there are particular subsets of patients in whom beating heart surgery can be particularly indicated and others for whom on-pump revascularization appears a better solution. Adaptation of the operation to the single patient is probably the way to improve outcome. PMID- 14759442 TI - Technical advances in total aortic arch replacement. AB - BACKGROUND: We compared the effects of using hypothermic circulatory arrest (HCA) alone, HCA combined with selective cerebral perfusion (SCP), and use of SCP with a trifurcated graft (T) on outcome after aortic arch repair. METHODS: One hundred fifty patients, median age 66 years (range, 27 to 85), underwent total arch replacement between 1988 and 2002; 75 were female. We retrospectively compared the results of three patient groups roughly comparable with regard to preoperative risk factors: 45 patients using HCA beginning in 1988; 67 patients using HCA/SCP beginning in 1994; and 38 patients utilizing a trifurcated arch graft in conjunction with SCP through the axillary artery (HCA/SCP/T) since 2000. The groups were well matched with regard to median age (66, 68, and 66 years), urgency (emergent 11%, 13%, 5%; urgent 24%, 9%, 18%; and elective 64%, 78%, 76%), and several other known risk factors (p = not significant). RESULTS: An adverse outcome-hospital death or permanent stroke-occurred in 14%: in 16% with HCA, in 16% with HCA/SCP, and in 8% with HCA/SCP/T. Transient neurologic dysfunction among patients surviving without stroke was lower with HCA/SCP/T (11%) than with HCA (33%) or HCA/SCP (17%). Mean duration of HCA was 52 +/- 16 minutes with HCA alone versus 45 +/- 10 minutes with HCA/SCP and 31 +/- 7 minutes with HCA/SCP/T (p < 0.0001 for groups HCA and HCA/SCP combined versus HCA/SCP/T). Mean duration of SCP was 57 +/- 25 minutes with HCA/SCP versus 62 +/- 24 minutes with HCA/SCP/T (p = not significant). Comparison of the groups of patients who had comparable preoperative risk factors for adverse outcome showed a trend toward lower adverse outcome and transient neurologic dysfunction rates using HCA/SCP/T; a significant reduction in respiratory (p < 0.001), infectious (p = 0.015) and cardiac (p = 0.005) complications in HCA/SCP/T compared with the earlier groups; and significantly shorter durations of intensive care (p < 0.0001) and hospitalization (p = 0.004). CONCLUSIONS: Our results suggest that HCA/SCP is superior to HCA alone for preventing cerebral injury during operations on the aortic arch. By further reducing embolic risk as well as duration of HCA, HCA/SCP/T with axillary artery cannulation may be the optimal technique for averting adverse outcomes, reducing complications, and shortening hospital stay after aortic arch repair. PMID- 14759443 TI - Thoracic aortic emergencies: impact of endovascular surgery. AB - BACKGROUND: Conventional surgery for thoracic aortic emergencies, such as contained or free rupture of thoracic aortic aneurysms, acute type B dissections, and traumatic rupture of the thoracic aorta, is frequently associated with a high rate of mortality and morbidity. To obviate this risk, endovascular surgery is considered to be a valid alternative procedure. METHODS: From March 2001 to July 2002, 15 of 22 patients with acute thoracic aortic syndromes were submitted to endovascular surgery: 3 patients (20%) for traumatic rupture, 4 patients (26.7%) for contained or free rupture of thoracic aortic aneurysm, and 8 patients (53.3%) for acute type B dissection evolving to rupture. Computed tomographic scan was diagnostic in all patients. The stent grafts were introduced through the femoral artery. RESULTS: In the endovascular group there were no perioperative deaths or open conversions. The intraoperative angiography and computed tomographic scan performed on discharge showed no significant endoleaks and successful sealing of the aortic dissection. Average intensive care unit and hospital stay was 1.7 +/- 0.8 and 5.9 +/- 3.0 days. Follow-up ranged between 4 and 23 months and included clinical examinations and serial computed tomographic scan at 3, 6, and 12 months, and every 6 months thereafter. One 84-year-old patient with thoracic aortic aneurysm died of pneumonia 78 days after endovascular surgery. Only one type 1 endoleak was noted in the first patient with traumatic rupture, 3 months after the procedure. CONCLUSIONS: Endovascular surgery is a safe technique, showing encouraging early and midterm results and allowing for prompt treatment of associated lesions in complex multitrauma patients. PMID- 14759444 TI - Predictors of cognitive decline following coronary artery bypass graft surgery. AB - BACKGROUND: A significant number of patients develop cognitive impairment that persists for months following coronary artery bypass grafting (CABG) surgery. Our objectives were to identify patient-related risk factors, processes of care, and the occurrence of any perioperative complications associated with cognitive decline. METHODS: Nine hundred thirty-nine patients enrolled in the Processes, Structures, and Outcomes of Care in Cardiac Surgery study undergoing CABG-only surgery at 14 Veterans Administration medical centers between 1992 and 1996 completed a short battery of cognitive tests at baseline and 6-months post-CABG. The composite cognitive score was based on the sum of errors for each individual item in the battery. Multiple linear regression analyses were used to identify independent predictors of the 6-month composite cognitive score. RESULTS: In multivariable analyses, patient characteristics associated with cognitive decline included cerebrovascular disease (p = 0.009), peripheral vascular disease (p = 0.007), history of chronic disabling neurologic illness (p = 0.016), and living alone (p = 0.049), while the number of years of education (p = 0.001) was inversely related to cognitive decline. After adjustment for baseline patient risk factors, the presence of any postoperative complication(s) (p = 0.001) was also associated with cognitive decline while cardiopulmonary bypass time (p = 0.008) was inversely related to cognitive decline. CONCLUSIONS: Patients with noncoronary manifestations of atherosclerosis, chronic disabling neurologic illness, or limited social support are at risk for cognitive decline after CABG surgery. In contrast, more years of education were associated with less cognitive decline. Preoperative assessment of risk factors identified in this study may be useful when counseling patients about the risk for cognitive decline following CABG surgery. PMID- 14759446 TI - Effects of hypertonic saline-dextran solution in cardiac valve surgery with cardiopulmonary bypass. AB - BACKGROUND: Hypertonic saline-dextran (HSD) solution may be beneficial in patients undergoing coronary artery surgery with cardiopulmonary bypass. Valvular dysfunction is associated with high pulmonary wedge pressure, pulmonary hypertension, and ventricular dysfunction. Fluid overload or transient left ventricular failure may occur with HSD infusion in such patients. This study evaluates the cardiorespiratory effects and tolerance of HSD solution infusion in patients undergoing cardiac valve surgery. METHODS: This prospective, randomized, double-blind study compared clinical, laboratory, hemodynamic, and respiratory measurements, and fluid balance in 50 patients over a 48-hour period after cardiopulmonary bypass for cardiac valve surgery. Twenty-five patients received 4 mL/kg of HSD during 20 minutes before cardiopulmonary bypass (HSD group). The control group received the same volume of Ringer's solution (Ringer group). RESULTS: Hospital mortality was zero. The HSD patients had a near zero fluid balance (6.5 +/- 13.5 mL/Kg/48 hours), and the control patients had a positive balance (91.0 +/- 33.7 mL/Kg/48 hours). Hemoglobin was similar in both groups, but more blood transfusions were necessary in the Ringer group (1.21 +/- 1.28 vs 0.48 +/- 0.59 units per patients). The HSD solution induced a higher cardiac index and left ventricular systolic work index postoperatively, and a lower systemic vascular resistance index until 6, 24, and 48 hours. Right ventricular systolic work index increased and pulmonary vascular resistance index decreased after HSD infusion. A better Pao(2)/Fio(2) relation was observed at 1 and 6 hours postoperatively in the HSD group and was associated with a shorter extubation time (432.0 +/- 123.6 vs 520.8 +/- 130.2 minutes). Increased oxygen delivery index occurred in the HSD group. The HSD infusion was well tolerated as none of the patients experienced fluid overload or had left ventricular failure develop. No other complication attributable to the use of HSD solution was observed. CONCLUSIONS: The HSD solution infusion in patients during cardiac valve surgery with cardiopulmonary bypass was well tolerated. Hemodynamic and respiratory functions improved and fluid balance was near zero during the first 48 hours as compared with a large positive balance in the control group. We conclude that HSD infusion is advantageous for patients undergoing cardiac valve surgery. PMID- 14759448 TI - Cardiopulmonary bypass in man: role of the intestine in a self-limiting inflammatory response with demonstrable bacterial translocation. AB - BACKGROUND: Cardiopulmonary bypass provokes a systemic inflammatory reaction that, in 1% to 2% of all cases, leads to multiorgan disfunction. The aim of this study was to evaluate the possible role of the intestine in the pathogenesis and development of this reaction. METHODS: Eleven selected patients scheduled for elective coronary artery bypass graft surgery were enrolled in a open, prospective clinical study. Gastric tonometry, chromium-labeled test and double sugar intestinal absorption tests, polymerase chain reaction microbial DNA test, and measurement of cytokines and transcriptional factor (nuclear factor kappaB) activation were performed. RESULTS: During the postoperative period, gastric pH remained stable (range,7.2 to 7.3). The partial pressure for carbon dioxide gradient between the gastric mucosa and arterial blood increased significantly (from 1 to 23 mm Hg), peaking in the sixth postoperative hour. Interleukin 6 increased significantly over basal levels, peaking 3 hours after cardiopulmonary bypass (96.3 versus 24 pg/mL). Nuclear factor kappaB never reached levels higher than those observed after lipopolysaccharide stimulation. Escherichia coli translocation was documented in 10 patients: in eight cases from removal of aortic cross-clamps and in two cases from the first postoperative hour. With respect to basal value (6.4%), the urine collection revealed a significant increase in excretion of the radioisotope during the first 24 hours after surgery (39.1%), although there were no significant variations with the double sugar test. CONCLUSIONS: The results obtained showed a correlation between the damage of the gastrointestinal mucosa, subsequent increased permeability, E coli bacteremia, and the activation of a self-limited inflammatory response in the absence of significant macrocirculatory changes and postoperative complications. PMID- 14759449 TI - Does off-pump CABG reduce gastrointestinal complications? AB - BACKGROUND: The aim of this study was to compare gastrointestinal complications and associated risk factors among patients undergoing cardiac surgery using off- and on-pump revascularization techniques. METHODS: A total of 1146 adult patients who underwent coronary artery surgery during a 6-year period were evaluated retrospectively. Group 1 consisted of 546 patients operated using off-pump techniques and group 2 consisted of 600 cases operated with cardiopulmonary bypass. Patients were compared and evaluated for gastrointestinal complications and possible associated risk factors using univariate and multivariate logistic regression analysis. RESULTS: Overall mortality was 1.6% in group 1 and 2.2% in group 2 (p = 0.523). Mortality due to gastrointestinal complications was 38.5% and 35.7% respectively in group 1 and group 2. The mean EuroSCORE value was 5.1 +/- 2.8 in group 1 and 3.8 +/- 2.4 in group 2 (p < 0.001). The most common gastrointestinal complication in the off-pump group was gastrointestinal bleeding. The leading complication in group 2 was intestinal ischemia. CONCLUSIONS: The incidence rates of gastrointestinal complications were similar in the on- and off-pump coronary artery bypass groups, the type of gastrointestinal complications, however, was different. Mortality rate due to these complications was also similar and remained high, regardless of the type of surgery. Cardiopulmonary bypass did not emerge as a risk factor for gastrointestinal complications, but prolonged cardiopulmonary bypass (longer than 98 minutes) resulted in a high incidence of such complications. Old age and advanced arteriosclerosis emerged as risk factors in both groups resulting in gastrointestinal complications suggesting the ischemic nature of the injury. PMID- 14759450 TI - Predictors of transfusion requirements for cardiac surgical procedures at a blood conservation center. AB - BACKGROUND: Previous studies defining perioperative risk factors for allogeneic transfusion requirements in cardiac surgery were limited to highly selected cardiac surgery populations or were associated with high transfusion rates. The purpose of this study was to determine perioperative risk factors and create a formula to predict transfusion requirements for major cardiac surgical procedures in a center that practices a multimodality approach to blood conservation. METHODS: We performed an observational study on 307 consecutive patients undergoing coronary artery bypass grafting, valve, and combined (coronary artery bypass grafting and valve) procedures. An equation was derived to estimate the risk of transfusion based on preoperative risk factors using multivariate analysis. In patients with a calculated probability of transfusion of at least 5%, intraoperative predictors of transfusion were identified by multivariate analysis. RESULTS: Thirty-five patients (11%) required intraoperative or postoperative allogeneic transfusions. Preoperative factors as independent predictors for transfusions included red blood cell mass, type of operation, urgency of operation, number of diseased vessels, serum creatinine of at least 1.3 mg/dL, and preoperative prothrombin time. Intraoperative factors included cardiopulmonary bypass time, three or fewer bypass grafts, lesser volume of acute normovolemic hemodilution removed, and total crystalloid infusion of at least 2,500 mL. The derived formula was applied to a validation cohort of 246 patients, and the observed transfusion rates conformed well to the predicted risks. CONCLUSIONS: A multimodality approach to blood conservation in cardiac surgery resulted in a low transfusion rate. Identifying patients' risks for transfusion should alter patient management perioperatively to decrease their transfusion rate and make more efficient use of blood resources. PMID- 14759451 TI - Cost analysis of aprotinin for coronary artery bypass patients: analysis of the randomized trials. AB - BACKGROUND: The full kallikrein-inhibiting dose of aprotinin has been shown to reduce blood loss, transfusion requirements, and the systemic inflammatory response associated with cardiopulmonary bypass graft surgery (CABG). A half-dose regimen, although having a reduced delivery cost, inhibits plasmin and fibrinolysis without substantially effecting kallikrein-mediated inflammation associated with bypass surgery. The differing pharmacologic effects of the two regimens impact the decision-making process. The current study assessed the medical cost offset of full-dose and half-dose aprotinin from short- and long term perspectives to provide a rational decision-making framework for clinicians. METHODS: To estimate CABG admission costs, resource utilization and clinical data from aprotinin clinical trials were combined with unit costs estimated from a Duke University-based cost model. Lifetime medical costs of stroke and acute myocardial infarction were based on previous research. RESULTS: Relative to placebo, the differences in total perioperative cost for primary CABG patients receiving full-dose or half-dose aprotinin were not significant. When lifetime medical costs of complications were considered, total costs in full-dose and half dose aprotinin-treated patients were not different relative to that of placebo. Total perioperative cost was significantly lower for repeat CABG patients treated with aprotinin, with savings of $2,058 for full-dose and $2,122 for half-dose patients when compared with placebo. Taking lifetime costs of stroke and acute myocardial infarction into consideration, the cost savings estimates were $6,044 for full-dose patients and $4,483 for half-dose patients, due to substantially higher lifetime stroke costs incurred by the placebo patients. CONCLUSIONS: Using this cost model, use of full-dose and half-dose aprotinin in primary CABG patients was cost neutral during hospital admission, whereas both dosing regimens were significantly cost saving in reoperative CABG patients. Additional lifetime cost savings were realized relative to placebo due to reduced complication costs, particularly with the full-dose regimen. As the full kallikrein-inhibiting dose of aprotinin has been shown to be safe and effective, the current results support its use in both primary and repeat CABG surgery. No demonstrable economic advantage was observed with the half-dose aprotinin regimen. PMID- 14759453 TI - Effects of nafamostat mesilate and minimal-dose aprotinin on blood-foreign surface interactions in cardiopulmonary bypass. AB - BACKGROUND: The pharmacological inhibition of blood-foreign surface interactions is an attractive strategy for reducing the morbidity associated with cardiopulmonary bypass. We compared the inhibitory effects of nafamostat mesilate (a broad-spectrum synthetic protease inhibitor) and minimal-dose aprotinin on blood-surface interactions in clinical cardiopulmonary bypass. METHODS: Eighteen patients undergoing coronary surgery were divided into three groups: (1) the control group (heparin, 4 mg/kg; n = 6), (2) the nafamostat mesilate group (heparin plus nafamostat, 0.2 mg/kg bolus followed by 2.0 mg/kg/h during cardiopulmonary bypass; n = 6), and (3) the aprotinin group (heparin plus aprotinin, 2.0 x 10(4) KIU/kg; n = 6). Platelet count, platelet aggregation, beta thromboglobulin, prothrombin fragment F1.2, thrombin-antithrombin complex, plasminogen activator inhibitor-1, alpha2-plasmin inhibitor-plasmin complex, D dimer, neutrophil elastase, and interleukin-6 were measured before, during, and after bypass. Bleeding times and blood loss were recorded. RESULTS: There were no significant differences between groups in platelet count, beta-thromboglobulin, plasminogen activator inhibitor-1, interleukin-6, bleeding times, or blood loss. Platelet aggregation was better preserved at 12 hours after surgery in the nafamostat and aprotinin groups than in the control group. Prothrombin fragment F1.2, thrombin-antithrombin complex and neutrophil elastase levels were significantly reduced by aprotinin, but not by nafamostat as compared with the control group. The alpha2-plasmin inhibitor-plasmin complex and D-dimer were significantly lower with either of the drugs. Aprotinin showed better control of D-dimer than did nafamostat. CONCLUSIONS: Nafamostat mesilate fails to reduce thrombin formation and neutrophil elastase release, whereas minimal-dose aprotinin inhibits both. Neither nafamostat nor aprotinin inhibits platelet activation. Nafamostat reduces fibrinolysis during cardiopulmonary bypass, although its effect is not as potent as aprotinin. PMID- 14759454 TI - Sodium-hydrogen exchange inhibition attenuates in vivo porcine myocardial stunning. AB - BACKGROUND: Inhibition of the sodium-hydrogen exchanger isoform 1 with HOE-642 (cariporide) has been shown to protect against ischemia-reperfusion injury and to decrease myocardial cell death in numerous animal preparations; however the effects of cariporide in stunned myocardium are not as well understood. We sought to determine whether cariporide attenuated myocardial stunning in vivo. METHODS: Open chest anesthetized pigs (22-33 kg) were subjected to 15 min of left anterior descending coronary artery (LAD) occlusion followed by 3 h of reperfusion. Regional ventricular function was assessed by segment shortening. Contractility was measured by stroke work and by load-insensitive preload recruitable stroke work and preload recruitable stroke work area. Vehicle or HOE-642 (1 mg/kg, IV) was administered 10 min before LAD occlusion. RESULTS: Cariporide treatment significantly improved postischemic segment shortening, stroke work, preload recruitable stroke work, and preload recruitable stroke work area and had no systemic hemodynamic effects. After 3 h of reperfusion, control animals recovered 33% +/- 4% and 33% +/- 3% of preischemic LAD segment shortening and preload recruitable stroke work area values, respectively, whereas animals treated with HOE-642 recovered 59% +/- 6% and 57% +/- 6%, respectively (p < 0.05). Seven (39%) of 17 control animals exhibited ventricular fibrillation during reperfusion; none of the cariporide-treated pigs fibrillated. CONCLUSIONS: Sodium-hydrogen exchange inhibition can attenuate postischemic myocardial stunning in addition to its well described anti-infarct properties. Inhibition of the sodium-hydrogen exchanger may be beneficial in patients susceptible to postischemic myocardial dysfunction associated with cardiac surgery. PMID- 14759455 TI - Effect of the Na+/H+ exchange inhibitor eniporide on cardiac performance and myocardial high energy phosphates in pigs subjected to cardioplegic arrest. AB - BACKGROUND: Pharmacologic Na(+)/H(+) exchange inhibition has been suggested to ameliorate cardiac performance depression associated with myocardial ischemia/reperfusion. The purpose of our experimental study was to investigate the impact of the novel Na(+)/H(+) exchange inhibitor Eniporide (EMD 96785) on cardiac performance and high energy phosphate content in a clinically relevant pig model of cardioplegic arrest. METHODS: We subjected 21 pigs (47 +/- 12 [SD] kg) to cardiopulmonary bypass (CPB) and 60 minutes cold (4 degrees C) crystalloid cardioplegic arrest (Bretschneider). The pigs were randomized to receive either systemic infusion of 3 mg/kg Eniporide before cardioplegia with added 2 micromol/L Eniporide (ENI-CP+iv; n = 7); 3 mg/kg Eniporide in cardioplegia only (ENI-CP; n = 7); or no Eniporide (control; n = 7). For cardiac performance determination we measured preload recruitable stroke work and Tau, the time constant of left ventricular (LV) isovolumic relaxation using sonomicrometry and micromanometry before CPB as well as 30, 60, and 120 minutes after weaning off CPB. LV and right ventricular myocardial adenine nucleotides (ATP, ADP, and AMP), glycogen, and water content were determined at the end of the experiments. RESULTS: Neither for standard hemodynamics including vascular pressures and cardiac index nor for cardiac performance factors did we find statistically significant differences between the groups. Similarly, myocardial adenine nucleotides, glycogene, and water content did not differ significantly between the groups. CONCLUSIONS: In this acute study we did not find significant effects of the Na(+)/H(+) exchange inhibitor Eniporide on cardiac performance and high energy phosphate content in healthy pig hearts subjected to ischemia/reperfusion induced by crystalloid cardioplegic arrest. PMID- 14759456 TI - Continuous cold blood cardioplegia improves myocardial protection: a prospective randomized study. AB - BACKGROUND: To assess the influence on myocardial protection of the rate of infusion (continuous vs intermittent) of cold blood cardioplegia administered retrogradely during prolonged aortic cross-clamping. The end-points were ventricular performance and biochemical markers of ischemia. METHODS: Seventy patients undergoing myocardial revascularization for three-vessel disease were prospectively randomized to receive intermittent or continuous retrograde cold blood cardioplegia. Hemodynamic measurements were obtained using a rapid-response thermodilution catheter and included right ventricular ejection fraction, cardiac output, left and right ventricular stroke work index, and systemic and pulmonary vascular resistance. Blood samples were obtained from the coronary sinus before cross-clamp application and immediately after cross-clamp removal for determinations of lactate and hypoxanthine. RESULTS: The left ventricular stroke work index trend was significantly superior (p = 0.038) by repeated-measures analysis in continuous cardioplegia. Other hemodynamic measurements revealed a similar trend. The need for postoperative inotropic drugs support was reduced in continuous cardioplegia. The release of lactate in the coronary sinus after unclamping was 2.30 +/- 0.12 mmol/L after intermittent cardioplegia and 1.97 +/- 0.09 mmol/L after continuous cardioplegia (p = 0.036). The release of hypoxanthine was 20.47 +/- 2.74 micromol/L in intermittent cardioplegia and 11.77 +/- 0.69 micromol/L in continuous cardioplegia (p = 0.002). CONCLUSIONS: Continuous cold blood cardioplegia results in improved ventricular performance and reduced myocardial ischemia in comparison with intermittent administration. PMID- 14759458 TI - Risk factors for mediastinitis after cardiac surgery. AB - BACKGROUND: Postoperative mediastinitis is one of the most feared complications in patients who undergo cardiac surgery because in addition to a high mortality rate (10% to 47%), there are increases in the length of hospital stay and in hospital costs. The purpose of the present study is to assess the risk factors for mediastinitis after cardiac surgery, the mediastinitis rate, and the mortality rate in our institution. METHODS: To determine the risk factors, a matched case-control study was carried out, with 39 cases and 78 controls, among the patients who underwent cardiac surgery at the Dante Pazzanese Cardiology Institute, Sao Paulo, Brazil. RESULTS: In the period of the study, 9,136 cardiac surgeries were performed and the mediastinitis rate was 0.5%. In the multivariate analysis, the independent risk factors found were obesity (odds ratio, 6.49; 95% confidence interval, 2.24 to 18.78), smoking (odds ratio, 3.27; 95% confidence interval, 1.04 to 10.20), intensive care unit stay more than 2 days (odds ratio, 4.50; 95% confidence interval, 1.57 to 12.90), and infection at another site (odds ratio, 8.86; 95% confidence interval, 1.86 to 42.27). The mortality rate was 23% among the patients with mediastinitis. CONCLUSIONS: We observed two independent risk factors related to patients' antecedents (obesity and smoking) and two risk factors related to problems in the postoperative period (length of intensive care unit stay and infection at another site). Efforts should be concentrated so that patients lose weight and stop smoking before elective surgeries. There should also be a prevention program against hospital infection directed to, and intensified for, at-risk patients. PMID- 14759457 TI - Superficial wound dehiscence after median sternotomy: surgical treatment versus secondary wound healing. AB - BACKGROUND: Superficial wound dehiscence after midline sternotomy is considered a minor complication in cardiac surgery, although it is quite frequent and requires prolonged medical treatment. It can be managed conventionally by topical treatment, with delayed secondary healing, or by surgical treatment and primary skin closure. We report the outcome of 96 patients who underwent conventional treatment, compared with a second group of 42 patients who underwent surgical treatment and direct closure. METHODS: From October 1999 to December 2002, 2400 consecutive patients underwent median sternotomy: 207 patients had sternal wound complications: 3 patients (0.125%) had mediastinitis, 66 patients (2.75%) had aseptic deep sternal wound dehiscence, and 138 patients (5.75%) had superficial wound dehiscence. The latter are the object of the present study; patients entered a protocol of skin wound care on an outpatient basis. The first 96 consecutive patients (group 1) required medications three times a week until complete healing. The last 42 patients (group 2) were treated by extensive surgical debridement of skin and subcutaneous tissue, direct closure of the superficial layers, and suture removal after 15 days. RESULTS: The two groups were comparable as to age, sex, and preoperative risk factors. The incidence of contaminated wounds was similar in the two groups (32 of 96 in group 1 and 11 of 42 in group 2; p = NS). The length of treatment was 29.7 days (range 2 to 144 days) for group 1 and 12.2 days (range 2 to 37 days) for group 2 (p < 0.0001). The mean number of medical treatments was 9.4 per patient in group 1 and 3.7 per patient in group 2 (p < 0.0001). CONCLUSIONS: Surgical debridement and primary closure of superficial surgical wound dehiscence after median sternotomy is a safe and valid treatment. Wound infection is not a contraindication to surgical treatment. Primary closure may contribute to reduce the risk for later infection. It also definitely contributes to decreasing healing time and strongly lessens patients' discomfort, diminishing hospital costs and hospital staff workload. PMID- 14759459 TI - Biomechanical study of a poly-L-lactide (PLLA) sternal pin in sternal closure after cardiothoracic surgery. AB - BACKGROUND: Stainless steel wiring is currently the standard method of median sternotomy closure but it has been reported that sufficient stiffness is not obtained by the method in anterior-posterior and cranial-caudal directions. A bioabsorbable Poly-L-Lactide (PLLA) sternal pin has been developed as an additive material for sternal closure. We biomechanically examined the effectiveness of a PLLA sternal pin in the two directions by using the sternum of a juvenile pig. METHODS: Juvenile pigs 14-17 kg weight were used. After the sternum was extirpated it was cut into two pieces at the midline. In a wire fixation group the pieces were fixed by two stainless wires. In a wire and intrasternal fixation group a hole was drilled into the bone marrow and a PLLA sternal pin for an infant was set into the hole. Then the both sides of the wired sternum were fixed tightly at the testing machine and the shear stress was forced into the one side. The shear stress was forced in anterior-posterior and cranial-caudal directions. RESULTS: In an anterior-posterior direction, the stiffness was 13.84 +/- 1.84 (N/mm) in a wire and intrasternal fixation group and 7.00 +/- 2.71 (N/mm) in a wire fixation group (p = 0.0002). In a cranial-caudal direction it was 10.61 +/- 4.88 (N/mm) and 4.38 +/- 2.12 (N/mm), respectively (p = 0.03). CONCLUSIONS: The use of a PLLA sternal pin as an additive material to steel wiring was effective in preventing the displacement of the sternum in both directions. Our data showed that closure technique using a sternal pin would provide adequate fixation. PMID- 14759460 TI - Off-pump mitral valve repair using the Coapsys device: a pilot study in a pacing induced mitral regurgitation model. AB - PURPOSE: The purpose of this study was to evaluate the ability of the Myocor Coapsys device to restore leaflet apposition and valve competency off-pump in a canine model of functional mitral regurgitation (MR). DESCRIPTION: The Coapsys device was surgically implanted in 10 dogs after MR induction by rapid ventricular pacing. The Coapsys consists of anterior and posterior epicardial pads connected by a subvalvular chord. The annular head of the posterior pad was positioned at the annular level to draw the posterior leaflet and annulus toward the anterior leaflet. Final device size was selected when MR was minimized or eliminated as assessed by color flow Doppler echocardiography. EVALUATION: All implants were placed off-pump without atriotomy, and mean MR grade was reduced from 2.9 +/- 0.7 to 0.6 +/- 0.7 (p < 0.001) acutely. No hemodynamic compromise was noted. CONCLUSIONS: The Coapsys device consistently and significantly reduced or eliminated functional MR acutely. Further study will be required to assess the chronic stability of the repair in this animal model. PMID- 14759462 TI - Successful treatment of massive pulmonary embolism in the 38th week of pregnancy. AB - In 1998, a 32-year-old woman in week 38 of pregnancy was referred to our hospital because of swelling of the lower extremities and dyspnea at rest. A massive pulmonary embolism and thrombi in the right and left atria were detected. After a successful cesarean section, emergent pulmonary embolectomy was performed with the use of cardiopulmonary bypass. Both the mother and the fetus recovered without severe complications. Postoperatively, activated protein S deficiency was detected. PMID- 14759463 TI - Pediatric thoracic lymphangiomatosis: is chest wall resection too radical? AB - Lymphangiomas account for 5% to 6% of all pediatric neoplasms. Complex pathologic processes often present a diagnostic and treatment challenge. Radical and major surgery might have a role in complex cases when medical treatment fails. PMID- 14759464 TI - Successful resuscitation of a patient with acute massive pulmonary embolism using emergent embolectomy. AB - Acute massive pulmonary embolism is associated with a high mortality rate. Prompt diagnosis and treatment are mandatory for a successful outcome. Although thrombolysis is effective, it is associated with a high rate of bleeding complications. This report describes the use of emergent pulmonary embolectomy as an effective and aggressive therapeutic approach to a massive saddle pulmonary embolism in a 66-year-old woman. With the application of specific surgical techniques and good interdisciplinary cooperation, pulmonary embolectomy may serve as more than a last resort for the management of this clinically unstable and dangerous condition. PMID- 14759465 TI - Recurrent intercostal herniation of the liver. AB - Abdominal intercostal hernia occurs rarely, with only 26 previous cases reported in the professional literature. A 51-year-old man presented with a painful right chest protrusion. One year earlier he had experienced a severe coughing spell and spontaneous rib fracture and chest protrusion. He was treated with endogenous tissue reinforcement and had no clinical improvement. Magnetic resonance imaging revealed the liver protruding through the chest wall. The hernia was reduced and the chest wall was repaired with prosthetic mesh and cables. Attention to the chest wall anatomy and reliable tissue closure including pericostal or transcostal nonabsorbable sutures and a prosthetic bridge over the defect are the best way to eliminate the patient's risk for recurrence. PMID- 14759466 TI - Migrating pleural mesothelial cyst. AB - We herein describe an atypical presentation of a migrating coelomic cyst attached to a pedicle of pericardial fat pad in an asymptomatic 45-year-old woman. A review of the English-language literature revealed only one such case report. PMID- 14759467 TI - Repair of stab-wound laceration of the aortic arch using deep hypothermia and circulatory arrest. AB - The surgical management of a 56-year-old patient with a single thoracic stab wound penetrating the left innominate vein and the aortic arch is described. Repair was successfully achieved using extracorporal circulation and circulatory arrest during deep hypothermia. Clinical features and surgical approach are described and discussed in detail. PMID- 14759468 TI - Role of Bartonella henselae endocarditis in the nucleation of aortic valvular calcification. AB - A 6-year-old boy presented with fatigability, shortness of breath, and bulging neck veins. Echocardiography revealed large vegetations, aortic insufficiency, a dilated left ventricle, and bicuspid aortic valve. There was no history of immunocompromise, fevers, or feline exposures. Blood cultures were negative; antibodies against Bartonella henselae were positive. Gentamicin was administered intravenously. Ross procedure was performed and patient was discharged on antibiotics in 5 days. Native valve was thickened by scar and fibrinous vegetations. Warthin-Starry stain demonstrated coccobacilli. Light and ultrastructural morphology, and monoclonal staining implicated B. henselae. Bacterial membranes contain calcium apatite crystals. Antigenic material was present in bacteria and calcified nodules. This case illustrates calcified protobacteria becoming incorporated into scar tissue during endocarditis. PMID- 14759469 TI - Dissection in a pulmonary autograft. AB - We report the case of a type II aortic dissection involving the pulmonary autograft after a Ross procedure 6 years earlier. A dissection flap was present in both the native ascending aorta and right coronary sinus of the autograft. At reoperation, the valve was spared using a root remodeling technique. PMID- 14759470 TI - Coronary endarterectomy and stent removal after iatrogenic perforation. AB - Coronary perforation is a rare complication of percutaneous coronary intervention. We report a case of type 3 coronary artery perforation after stenting of the left anterior descending coronary artery. Pericardiocentesis was required to treat cardiac tamponade and prolonged balloon inflation did not stop the bleeding. Urgent surgical intervention with coronary endarterectomy, removal of the stent, and bypass grafting using the left internal mammary artery to the left anterior descending artery was successful. Complete removal of stent with endarterectomy is a feasible option for perforation as a complication of coronary stenting. PMID- 14759471 TI - Failure of argatroban anticoagulation during off-pump coronary artery bypass surgery. AB - Heparin-induced antibodies create vexing problems during cardiac surgery. Although alternative medications have been used for intraoperative anticoagulation, the results have been sufficiently variable that no one medication is recommended. In our case, due to the poor reversibility of the antithrombin agents, argatroban was chosen as a heparin substitute due to its short half-life and its anticoagulation assessment using the activated clotting time (ACT). Unfortunately, our experience was that argatroban does not provide adequate anticoagulation during off-pump coronary bypass surgery, even when the ACT is maintained at more than 380 sec. PMID- 14759472 TI - Accessory mitral valve tissue causing severe left ventricular outflow tract obstruction in an adult. AB - Accessory mitral valve (AMV) is a rare cause of left ventricular outflow tract (LVOT) obstruction and is extremely rare in adults. We report a case of an older adult with an AMV that caused severe LVOT obstruction. A parachute-like piece of tissue (the AMV) protruding into the LVOT during systole was first detected in a 45-year-old woman by echocardiography. Because the pressure gradient and dyspnea gradually progressed, she finally underwent a successful operation for removal when she was 48 years old. PMID- 14759473 TI - Successful salvage of inferior vena cava rupture during reoperative repair for ascending aortic aneurysm. AB - A 44-year-old man underwent reoperative repair for ascending aortic aneurysm at our hospital. He had received aortic valve replacement due to severe aortic regurgitation 4 years previously. The cardiopulmonary bypass was set up through the femoral arterial and venous cannula. An unusual experience of rupture of the abdominal inferior vena cava was encountered after total-body retrograde perfusion. The tragic situation was successfully salvaged by recannulation into the intrapericardial inferior vena cava and repair of the several perforations on abdominal inferior vena cava. The patient recovered smoothly except for mild pancreatitis. The causes of rupture of the inferior vena cava are discussed here. PMID- 14759474 TI - Aortic valve replacement in a patient with a patent internal thoracic artery graft. AB - Myocardial protection in patients requiring a second open-heart surgical procedure after coronary artery bypass grafting, especially when there is a patent left internal thoracic artery graft to the left anterior descending coronary artery, remains controversial. We present the case of a patient in whom aortic valve replacement was undertaken 18 months after coronary artery revascularization. Unusual features included beating-heart aortic valve replacement with continuous retrograde coronary sinus perfusion and avoidance of dissection of the patent grafts, including the left internal thoracic artery and a saphenous vein graft. PMID- 14759475 TI - Successful biventricular assistance after heart transplantation. AB - We present a successful use of biventricular pneumatic assistance during seven days allowing the recovery from a severe primary graft failure that occurred in the operating room. Suboptimal donors are associated with higher rates of immediate graft failure; however proper use of mechanical assistance can help to recover graft function. PMID- 14759476 TI - Left atrial appendage aneurysm in a child. AB - We report a case of left atrial appendage aneurysm in a 1-year-old child. The patient was asymptomatic, and a mediastinal liquid mass that was confirmed to be an aneurysm of the left atrial appendage was incidentally revealed by transthoracic echocardiography. Aneurysmectomy was performed during cardioplegic arrest under the support of cardiopulmonary bypass. The postoperative course was uneventful. PMID- 14759477 TI - Intrathoracic parathyroid adenoma. PMID- 14759478 TI - Elective surgery for fibroelastoma of the aortic valve. PMID- 14759479 TI - Uniportal VATS wedge pulmonary resections. AB - Since 2000, 15 patients have undergone single port (uniportal) video-assisted thoracic surgery for wedge pulmonary resection either for diagnosis of interstitial lung diseases (10 patients) or for treatment of primary spontaneous pneumothoraces (5 patients). Diagnosis was obtained in all patients and no recurrences of pneumothorax were seen at follow-up. This initial experience shows that, for specific indications, uniportal video-assisted thoracic surgery for wedge pulmonary resections can be safe and effective. PMID- 14759480 TI - Posterior mitral valve restoration for ischemic regurgitation. AB - Chronic ischemic mitral regurgitation is traditionally a complex lesion to repair. Only restrictive annuloplasty has become an accepted strategy to avoid valve replacement, but results are unsatisfactory in some subgroups of patients. We describe an original technique that addresses the pathophysiologic mechanisms responsible for one of the most common subtypes of ischemic mitral regurgitation, ie, asymmetric tethering of the mitral leaflets after inferior myocardial infarction. The technique includes partial detachment of the posterior leaflet from the mitral annulus, annular plication, and posterior cusp plasty. PMID- 14759481 TI - Optimizing use of the octopus system for off-pump total arterial myocardial revascularization with the TY graft. AB - Rapid developments in interventional cardiology for the treatment of coronary artery disease ask for further evolution of surgical revascularization techniques with improved short-term and long-term results. Off-pump bypass grafting, total arterial revascularization, and use of composite arterial grafts are innovative operative procedures that are already established in many centers. We describe our technique to combine all three procedures in a single operation. PMID- 14759482 TI - Autologous repair of supravalvar pulmonic stenosis. AB - Native supravalvar pulmonary stenosis is a rare anomaly, but iatrogenic supravalvar pulmonary stenosis occurs after various repairs for congenital heart disease with relative frequency. Surgical techniques such as patching carry the risk of restenosis. We describe a technique of repair using only autologous tissues that can be applied to both native and iatrogenic supravalvar pulmonary stenosis. There were no complications and no patient developed restenosis at follow-up. Autologous repair of supravalvar pulmonary stenosis is an effective technique. PMID- 14759483 TI - Immunosuppression and xenotransplantation of cells for cardiac repair. AB - The death of highly vulnerable cardiomyocytes during ischemia leads to cardiac dysfunction, including heart failure. Due to limited proliferation of adult mammalian cardiomyocytes, the dead myocardium is replaced by noncontractile fibrotic tissue. Introducing exogenous cells to participate in the regeneration of infarcted myocardium has thus been proposed as a novel therapeutic approach. In view of the availability of various xenogeneic cells and fewer ethical and political concerns that surround human embryonic stem cells and fetal cardiomyocytes, cellular xenotransplantation may be a potential alternative approach for cardiac repair in humans. However, one of the most daunting challenges of xenotransplantation is immunorejection. This article summarizes the progress in cellular xenotransplantation for cardiac repair in experimental settings and the current understanding of possible immune responses following the engraftment of xenogeneic cells. The public attitude towards xenotransplantation is reportedly more favorable to receiving cells or tissues than a whole organ, but many scientific obstacles need to be overcome before the utilization of xenogeneic cells for cardiac repair in patients with heart disease becomes applicable to clinical practice. PMID- 14759484 TI - Off-pump myocardial revascularization is associated with less incidence of stroke in elderly patients. AB - Several recent studies have highlighted the potential benefits of using off-pump coronary artery bypass (OPCAB) surgery, particularly in high-risk patients. The aim of this meta-analysis is to assess the effect of OPCAB on the incidence of stroke compared with coronary artery bypass grafting using cardiopulmonary bypass (CPB) in elderly patients. We performed a meta-analysis of all observational studies, published in MEDLINE between 1999 and 2002 and a comparison between the OPCAB and CPB techniques in elderly patients was performed with the outcome of interest being the incidence of stroke. Elderly patients were defined as those aged 70 years or older. Nine studies are included in the meta-analysis. The total number of subjects included was 4,475 patients, of which, 1,253 underwent OPCAB (28%) and 3,222 (72%) underwent CPB. The meta-analysis showed that the OPCAB technique was associated with significantly lower incidence of stroke in elderly patients compared with the CPB technique (1% vs 3%), with an odds ratio of 0.38% to 95% (CI, 0.22 to 0.65). We did not identify any significant heterogeneity and funnel plot asymmetry between the studies included in the meta-analysis. Meta regression analysis including variables predicting stroke, mortality, and study characteristics did not show any associations affecting the calculated odds ratio of stroke. Despite the fact that this is a meta-analysis of observational studies and adjustment for differences in baseline risk factors between OPCAB and CPB patients was not possible, we believe that this study suggests that the OPCAB technique might be associated with reduced incidence of stroke in the elderly patients undergoing coronary artery bypass grafting. PMID- 14759485 TI - Blunt chest trauma and tube thoracostomy. PMID- 14759487 TI - Discrepancy in serum carcinoembryonic antigen levels by measuring kits. PMID- 14759489 TI - Cefazolin prophylaxis during cardiac operations. PMID- 14759491 TI - Statue with clubbed feet: another reading. PMID- 14759493 TI - Involvement of increased stability of mitochondrial membrane potential and overexpression of Bcl-2 in enhanced anoxic tolerance induced by hypoxic preconditioning in cultured hypothalamic neurons. AB - The effects of hypoxic preconditioning (HP) on changes in mitochondrial membrane potential (MMP) and Bcl-2 expression in cultured hypothalamic neurons after severe anoxia were investigated. In the HP group, hypothalamic neurons, after a 4 day culture, were preconditioned daily under a hypoxic condition (1% O(2), 10 min) for 8 days; subsequently, the HP neurons and those in the control group (similarly cultured, but without HP) were exposed to 6 h of severe anoxia (0% O(2)). The preconditioned neurons had a higher survival rate and a lower lactate dehydrogenase leakage, compared with the control group. Although HP did not prevent the degradation of MMP during severe hypoxia, preconditioned neurons exhibited a higher level of MMP than that of the control group. Increased expression of Bcl-2 was also observed in the preconditioned hypothalamic neurons. These results suggest that HP enhances the hypoxic tolerance of hypothalamic neurons, and the underlying mechanisms may be related to the increased stability of MMP and the overexpression of Bcl-2 induced by HP. PMID- 14759494 TI - Post-ischemic delayed expression of hepatocyte growth factor and c-Met in mouse brain following focal cerebral ischemia. AB - We investigated long-term changes in the expression of protein and mRNA of hepatocyte growth factor (HGF) and its receptor c-Met in mouse brain after permanent occlusion of the middle cerebral artery, by using immunohistochemistry and quantitative reverse transcription-polymerase chain reaction. HGF immunopositive cells were observed in the periinfarct region from 4 days after occlusion, peaking at 14-28 days. The area containing HGF-immunopositive cells continued to expand until 28 days after occlusion. c-Met-immunopositive cells were observed exclusively at the periinfarct region at 7 and 14 days after occlusion. At 28 days after occlusion, there were many c-Met-immunopositive cells in the widespread periinfarct region. Triple immunohistochemical staining by using confocal laser scanning microscopy (CLSM) demonstrated that most of the HGF immunopositive cells were localized to reactive astrocytes. The c-Met immunopositive cells were also localized to reactive astrocytes. HGF mRNA was upregulated exclusively in the periinfarct region at 14 days. c-Met mRNA was upregulated in the periinfarct region from as late as 28 days after occlusion. Thus, HGF and c-Met show delayed expression in the periinfarct region at both protein and mRNA levels after induction of ischemia. Because HGF was recently shown to play critical roles in angiogenesis and neurotrophic activities, the temporal profiles of their expression may imply the involvement of HGF in the process of post-ischemic brain tissue repair. PMID- 14759495 TI - Compromised metabolic recovery following spontaneous spreading depression in the penumbra. AB - Spreading depression (SD) has been demonstrated following focal ischemia, and the additional workload imposed by SD on a tissue already compromised by a marked reduction in blood flow may contribute to the evolution of irreversible damage in the ischemic penumbra. SD was elicited in one group of rats by injecting KCl directly into a frontal craniectomy and the wave of depolarization was recorded in two craniectomies 3 and 6 mm posterior to the first one. In a second group, the middle cerebral artery was occluded using the monofilament technique and a recording electrode was placed 5 mm lateral to the midline and 0.2 mm posterior to bregma. To determine the metabolic response in the penumbral region of the cortex ipsilateral to the occlusion, brains from both groups were frozen in situ when the deflection of the SD was maximal. The spatial metabolic response of SD in the ischemic cortex was compared to that in the non-ischemic cortex. Coronal sections of the brains were lyophilized, pieces of the dorsolateral cortex were dissected and weighed, and analyzed for ATP, P-creatine, inorganic phosphate (Pi), glucose, glycogen and lactate at varying distances anterior and posterior to the recording electrode. ATP and P-creatine levels were significantly decreased at the wavefront in both groups and the levels recovered after passage of the wavefront in the normal brain, but not in the ischemic brain. Glucose and glycogen levels were significantly decreased and lactate levels significantly increased in the tissue after the passage of the wavefront. While the changes in the glucose-related metabolites persisted during recovery even in anterior portions of the cortex in both groups in the aftermath of the SD, the magnitude of the changes was greater in the penumbra than in the normal cortex. SD appears to impose an equivalent increase in energy demands in control and ischemic brain, but the ability of the penumbra to recover from the insult is compromised. Thus, increasing the energy imbalance in the penumbra after multiple SDs may hasten the deterioration of the energy status of the tissue and eventually contribute to terminal depolarization and cell death, particularly in the penumbra. PMID- 14759496 TI - The phototransduction cascade in the isolated chick pineal gland revisited. AB - It is well established that the isolated chick pineal gland is directly light sensitive and that melatonin synthesis of the gland can be inhibited by exposing the gland to light during scotophase. Since not all the steps of the phototransduction cascade have been clarified to the same extent as in the retina, we have treated isolated chick pineal glands with 90 min of light during scotophase and with drugs that affect key-components of vertebrate phototransduction, i.e., cyclic guanosine monophosphate (cGMP) phosphodiesterase 6 (PDE6), cGMP levels and cGMP-gated calcium channels. The endpoint measured was the activity of the rate-limiting enzyme of melatonin synthesis, arylalkylamine N acetyltransferase (AA-NAT), which is inhibited by light. The effects on AA-NAT activity of light were negated by addition of dipyridamol and zaprinast, either of which inhibits the light-induced activation of PDE6. The effect of light was also counteracted by the nitric oxide donor sodium nitroprusside and C-type natriuretic peptide, both of which increase cGMP levels, and by the calcium channel agonist Bay K 8644, which prevents the cGMP-decrease-induced closure of cGMP-gated calcium channels. Inhibition of nitric oxide synthase (NOS) by N(G) nitro-l-arginine did not influence the inhibitory effect of light, suggesting that the NOS pathway does not play a role. Since the light effect on AA-NAT activity involves both cGMP and cyclic adenosine monophosphate (cAMP) hydrolysis, we have also studied whether the cGMP-inhibited cAMP phosphodiesterase 3 (PDE3) is involved. As the specific PDE3 inhibitor cilostamide is without effect, we assume that the light-induced decrease of cAMP levels does not involve PDE3. These results taken together strongly suggest that the investigated steps of the phototransduction cascade in the isolated chick pineal gland are basically similar to those in the retina. PMID- 14759497 TI - Central administration of cocaine- and amphetamine-regulated transcript increases phosphorylation of cAMP response element binding protein in corticotropin releasing hormone-producing neurons but not in prothyrotropin-releasing hormone producing neurons in the hypothalamic paraventricular nucleus. AB - Cocaine- and amphetamine-regulated transcript (CART) has an important action on hypophysiotropic thyrotropin-releasing hormone (TRH) and corticotropin-releasing hormone (CRH) neurons to regulate the hypothalamic-pituitary-thyroid and adrenal axis, respectively. To elucidate the mechanisms by which CART mediates its effect on TRH and CRH neurons, we determined whether the exogenous administration of CART into the cerebrospinal fluid (CSF) phosphorylates the transcription factor, cyclic adenosine 5'-monophosphate response element binding protein (CREB), in the nucleus of TRH and CRH neurons. CART dramatically increased the percentage of phosphoCREB (PCREB) immunolabeled cell nuclei in the hypothalamic paraventricular nucleus (PVN) in fasted as well as fed rats at 10-min postinjection, particularly in the medial parvocellular subdivision of the PVN. Double immunolabelling with CRH antiserum revealed that CART increased the number of CRH neurons containing PCREB from 10.5+/-1.2 % to 87+/-1.2% (P<0.001) in fasting animals and from 3.7+/ 0.8% to 74+/-5.3% (P<0.001) in fed animals. In contrast, no significant change was observed in the percentage of proTRH neurons colocalizing with PCREB either in the fasted (11.7+/-1.85%) or fed animals (4.2+/-2.2%) as compared to their respective vehicle controls (2.5+/-1.4% and 4.6+/-1%). Ultrastructural analysis revealed that CART establishes axosomatic and axodendritic contacts with CRH neurons in the PVN. These data demonstrate a selective effect of CART to phosphorylate CREB in CRH, but not TRH neurons in the PVN. Since CART is capable of increasing the gene expression of both CRH and TRH in hypophysiotropic neurons, and CART-containing axon terminals establish synaptic relationships with hypophysiotropic CRH and TRH neurons, we propose that CART may signal to the nucleus by more than one pathway. PMID- 14759498 TI - Differences in alpha7 nicotinic acetylcholine receptor binding in motor symptomatic and asymptomatic MPTP-treated monkeys. AB - We studied [(125)I]alpha-bungarotoxin (btx) binding to alpha7 nicotinic acetylcholine receptors in normal and 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP) exposed macaque monkeys. [(125)I]alpha-Btx binds throughout the normal monkey brain, with the greatest density in the thalamic nuclei and with moderate to low binding in the hippocampus, prefrontal cortex, caudate, putamen, and substantia nigra. Chronic administration of low doses of MPTP resulted in animals with stable cognitive deficits without overt parkinsonian motor symptoms. [(125)I]alpha-Btx binding in the brains of these animals was significantly increased in the outermost layers of the supplementary motor cortex (area 6M, approximately 50%), primary motor cortex (area 4, approximately 112%) and throughout the putamen (approximately 50-72%). In contrast, there was no change in [(125)I]alpha-btx binding in the brain regions thought to be involved in mediating the cognitive functions impaired in these monkeys (e.g., the hippocampus, areas 9/46D and 46D of the principal sulcus, and area 24c of the cingulate sulcus). Animals with cognitive dysfunction that received escalating doses of MPTP for >6 months developed motor signs of parkinsonism which were indistinguishable from those seen in animals rendered acutely parkinsonian with short term administration of large doses of MPTP. These two "motor symptomatic" groups had significantly increased [(125)I]alpha-btx binding only in the dorsolateral putamen. Immunohistochemical studies showed that the increased [(125)I]alpha-btx binding, when observed, was associated with enhanced immunohistochemical staining localized to neurons and was not a result of an astrocytic response to MPTP. These results suggest that the increase in alpha7 nicotinic acetylcholine receptor expression in the chronic low-dose MPTP treated, motor asymptomatic monkeys may be a part of compensatory processes contributing to the maintained motor functioning in these animals. PMID- 14759499 TI - Expression of B/K protein in the hippocampus of kainate-induced rat seizure model. AB - B/K protein is a newly identified member of double C2-like domain protein family. We examined the expression of B/K protein in the hippocampus of kainate-induced rat seizure model. Intraperitoneal injection of kainate increased the immunoreactivity to B/K protein in the CA1 to CA3 of the hippocampus. B/K protein expression began to increase at 6 h, reached the maximum at 12 h, and then returned nearly to the normal level at 72 h after the injection of kainate (12 mg/kg), and it was also dependent on the dose of kainate between 4 and 16 mg/kg. In electron microscopic and subcellular fractionation studies, B/K protein was localized in the endoplasmic reticulum (ER) of the hippocampus. Kainate also induced the expression of BiP, a typical ER stress marker protein, in the hippocampus and the cortex, and it was coexpressed with B/K protein. Moreover, thapsigargin-induced ER stress caused upregulation of B/K protein expression in PC12 cells. In conclusion, our data showing the induction of both B/K protein expression and ER stress response in the hippocampus of kainate seizure model, and ER-specific expression and ER stress-induced expression of B/K strongly suggest the possible role of B/K protein in epileptogenesis or epilepsy-induced neuronal damage. PMID- 14759500 TI - Neuroprotective effect of dapsone in an occlusive model of focal ischemia in rats. AB - Dapsone (4,4'-diaminodiphenylsulfone) is employed in the chemotherapy against leprosy. Dapsone also prevents neuronal damage induced by glutamate agonists. As glutamate excitotoxicity is implicated in the damage after ischemia, we tested the ability of dapsone to prevent ischemic injury in a model of permanent middle cerebral artery (MCA) occlusion in rats. Either dapsone (9.375 or 12.5 mg/kg doses) or vehicle were i.p. administered 30 min after occlusion. Rats from the control group showed a permanent neurological deficit after occlusion, while dapsone-treated groups improved significantly. Dapsone-treated animals showed a reduction of 93% (9.375 mg/kg dose) and 92% (12.5 mg/kg dose) in the infarction volume as compared to control values. PMID- 14759501 TI - Embryonic stem cells are capable of generating a neuronal network in the adult mouse retina. AB - The integration of embryonic stem (ES) cells as well-differentiated neuronal cells into the retinas of adult mice was investigated. ES cells were transplanted in adult mouse retinas by intravitreal injections. Neuronal differentiation of the ES cells was investigated by morphological and immunohistochemical examinations on post-operative days 5 and 30. ES cell apoptosis was examined by in situ terminal dUTP-biotin nick end labeling of DNA fragments. Differentiated ES cells growing along the retinal surface developed fine neuronal cell processes around cell nuclei and generated neuronal networks into the retinal inner plexiform layer (IPL) 30 days after transplantation. The differentiated ES cells expressed retinal and neuronal markers. Many apoptotic cells were recognized in transplanted ES cells at day 5 but not at day 30 after transplantation. ES cells may be useful for neural tissue regeneration in the adult mammalian retina. PMID- 14759502 TI - Heat-shock pretreatment prevents suppression of long-term potentiation induced by scopolamine in rat hippocampal CA1 synapses. AB - We examined the effect of heat-shock pretreatment on long-term potentiation (LTP) in the CA1 hippocampal slices of the rat using the muscarinic blocker scopolamine as the LTP (memory) suppressor. Time course study using immunohistochemical techniques indicated peak expression of HSP70 16 h after heat-shock treatment. Focusing on that time point we found tetanic stimulation (at 100 Hz) induced LTP of 191.1+/-12.2% in control slices (n=7), which was suppressed by scopolamine to 114.5+/-2.8 %. Heat-shock pretreatment successfully prevented such suppression (216.6+/-38.2% and 190.2+/-10.6% with and without scopolamine, respectively, n=7). Both HSP expression and LTP responses were relatively small taken either 2 or 48 h after heat-shock or sham pretreatment. These results suggest that the induction of HSPs is time-dependent and can prevent scopolamine-mediated LTP suppression. PMID- 14759503 TI - Synergistic effects of cannabinoid inverse agonist AM251 and opioid antagonist nalmefene on food intake in mice. AB - Oral administration of the opioid antagonist nalmefene alone (up to 20 mg/kg) failed to show a significant effect on acute food intake in mice. However, combined oral dosing of nalmefene and subthreshold doses of AM251, a cannabinoid CB1 receptor inverse agonist, led to a significant reduction in food intake in both lean and diet-induced obese (DIO) mice. Furthermore, the anorectic effect of a high dose of AM251 was further enhanced when co-administered with nalmefene. The results support a synergistic interaction between opioid and cannabinoid systems in regulating feeding behavior. PMID- 14759504 TI - Region-specific alterations of neuronal nitric oxide synthase (nNOS) expression in the amygdala of the aged rats. AB - Although many amygdalar functions are altered by aging, little is known about their mechanisms. As these functions are related with nitric oxide (NO), we examined neuronal nitric oxide synthase (nNOS) expression in the amygdala of the aged rats via immunohistochemical technique. We found that nNOS immunoreactive neurons are decreased in almost all amygdalar areas of the aged rats, while nNOS immunoreactivity of the neuropil is significantly increased in the amygdalar nuclei related with main and accessory olfactory system. These suggest altered levels of NO might provide region-specific mechanisms of many physiological and behavioral deficits of the amygdala developed by aging. However, exact effects of these changes require further elucidation. PMID- 14759507 TI - Highly efficient renaturation of beta-lactamase isolated from moderately halophilic bacteria. AB - Most, if not all, beta-lactamases reported to date are irreversibly denatured at 60-70 degrees C. Here, we found that a halophilic beta-lactamase from the moderately halophilic bacterium Chromohalobacter sp. 560 was highly stable against heat inactivation: it retained approximately 75% of its activity after boiling for 5 min in the presence of 0.2 M NaCl, suggesting that the protein either incompletely denatures during the boiling process or readily renatures upon cooling to the assay temperature. Circular dichroism showed a complete unfolding at 60 degrees C and a full reversibility, indicating that the observed activity after boiling is due to efficient refolding following heat denaturation. The enzyme showed optimal activity at 50-60 degrees C, indicating that an increase in activity with temperature offsets the thermal denaturation. The gene bla was cloned, and the primary structure of the enzyme was deduced to be highly abundant in acidic amino acid residues, one of the characteristics of halophilic proteins. Despite its halophilic nature, the enzyme refolds in low salt media after heat denaturation. PMID- 14759508 TI - Location of the Escherichia coli RNA polymerase alpha subunit C-terminal domain at an FNR-dependent promoter: analysis using an artificial nuclease. AB - The Escherichia coli FNR protein is a global transcription regulator that activates gene expression via interactions with the RNA polymerase alpha subunit C-terminal domain. Using preparations of E. coli RNA polymerase holoenzyme, specifically labelled with a DNA cleavage reagent, we have determined the location and orientation of the C-terminal domain of the RNA polymerase alpha subunit in transcriptionally competent complexes at a class II FNR-dependent promoter. We conclude that one alpha subunit C-terminal domain binds immediately upstream of FNR, and that its position and orientation is the same as at similar promoters dependent on CRP, another E. coli transcription activator that is related to FNR. In complementary experiments, we show that the second alpha subunit C-terminal domain of RNA polymerase can be repositioned by upstream-bound CRP, but not by upstream-bound FNR. PMID- 14759509 TI - Increased transcription of mitochondrial genes for Complex I in human platelets during ageing. AB - We studied the effect of ageing on the mRNA levels of mitochondria-encoded polypeptides in human platelets. We used quantitative real-time reverse transcriptase-polymerase chain reaction (RT-PCR) to investigate the expression of selected cytochrome c oxidase (COX) genes (subunits I and III) and Complex I genes (subunits reduced nicotinamide adenine dinucleotide (NADH) dehydrogenase (ND)1 and (ND)5 in platelets from young and aged healthy subjects. Northern blot analysis confirmed the PCR results. COX I expression is higher than that of COX III in both young and aged platelets. A significant increase of transcripts for Complex I was found during ageing. On the contrary, the mRNA levels of the two COX subunits did not significantly vary during ageing. PMID- 14759510 TI - Revisiting the odorant-binding protein LUSH of Drosophila melanogaster: evidence for odour recognition and discrimination. AB - LUSH is a soluble odorant-binding protein of the fruit fly Drosophila melanogaster. Mutants not expressing this protein have been reported to lack the avoidance behaviour, exhibited by wild type flies, to high concentrations of ethanol. Very recently, the three-dimensional structure of LUSH complexed with short-chain alcohols has been resolved supporting a role for this protein in binding and detecting small alcohols. Here we report that LUSH does not bind ethanol and that wild type flies are in fact attracted by high concentrations of ethanol. We also report that LUSH binds some phthalates and that flies are repelled by such compounds. Finally, our fluorescence data, interpreted in the light of the three-dimensional structure of LUSH, indicate that the protein undergoes a major conformational change, similar to that reported for the pheromone-binding protein of Bombyx mori, but triggered, in our case, by ligand. PMID- 14759511 TI - Growth hormone is a positive regulator of adiponectin receptor 2 in 3T3-L1 adipocytes. AB - The fat-derived protein adiponectin is an important insulin-sensitizing adipocytokine which is downregulated in insulin resistance and obesity. Recently, two receptors of this adipose-expressed protein called adiponectin receptor 1 (AdipoR1) and 2 (AdipoR2) have been cloned. To clarify expression and regulation of these receptors in fat cells, AdipoR1 and AdipoR2 mRNA was measured by quantitative real-time reverse transcription-polymerase chain reaction during differentiation of 3T3-L1 adipocytes and after treatment with various hormones known to induce insulin resistance. Interestingly, AdipoR2 synthesis was significantly increased up to 4.8-fold during differentiation of 3T3-L1 preadipocytes, whereas AdipoR1 expression was only augmented up to 1.4-fold. Furthermore, growth hormone (GH) induced AdipoR2, but not AdipoR1 mRNA by up to 2.4-fold in a dose- and time-dependent fashion with significant stimulation detectable at concentrations as low as 5 ng/ml GH and as early as 2 h after effector addition. The positive effect of GH on AdipoR2 expression could be reversed by withdrawal of the hormone for 24 h. In contrast, other key hormones involved in the regulation of insulin resistance and energy metabolism such as insulin, isoproterenol, dexamethasone, triiodothyronine, angiotensin 2, tumor necrosis factor alpha, and interleukin-6 did not influence AdipoR1 and AdipoR2 synthesis in vitro. Taken together, our results suggest that AdipoR2 expression is differentiation-dependent and selectively regulated by GH implying a potential role of this hormone in adiponectin-associated alterations of insulin sensitivity and energy homeostasis. PMID- 14759512 TI - PPARgamma mediates NSAIDs-induced upregulation of TFF2 expression in gastric epithelial cells. AB - Trefoil factor family (TFF) is a group of peptides that play critical roles in maintaining gastric mucosal integrity. In real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR) and reporter gene assays, we show that indomethacin and aspirin upregulate TFF2 expression in MKN45 gastric cells. These drugs also activated peroxisome proliferator-activated receptor gamma (PPARgamma) at concentration ranges that increase TFF2 expression, and upregulated TFF2 expression was suppressed by GW9662, a specific inhibitor of PPARgamma. These results suggest that indomethacin and aspirin upregulate gastric expression of TFF2 through activation of PPARgamma. This mechanism may be important in reducing the extent of gastric mucosal injury caused by the administration of non-steroidal anti-inflammatory drugs (NSAIDs). PMID- 14759513 TI - Functional properties of soybean nodulin 26 from a comparative three-dimensional model. AB - A model of the nodulin 26 channel protein has been constructed based on comparative modeling and molecular dynamics simulations. Structural features of the protein indicate a selectivity filter that differs from those of the known structures of Escherichia coli glycerol facilitator and mammalian aquaporin 1. The model structure also reveals important roles of Ser207 and Phe96 in ligand binding and transport. PMID- 14759514 TI - Mutational analysis of GstI protein, a glutamine synthetase translational inhibitor of Rhizobium leguminosarum. AB - The small GstI protein (63 amino acids) of Rhizobium leguminosarum inhibits the expression of the glnII (glutamine synthetase II) gene, thus reducing the bacterial ability to assimilate ammonium. In order to identify the residues essential for its inhibitory activity, all the 53 non-alanine amino acid residues of GstI were individually mutated into alanine. Based on their capacity to inhibit glnII expression (in two genetic backgrounds) three groups of mutants were identified. The first group displayed an inhibitory activity similar to the wild-type; the second and the third ones showed partial and total loss of inhibitory activity, respectively. Several mutations of the latter group concerned residues conserved in two related sequences from Sinorhizobium meliloti and Agrobacterium tumefaciens. Additionally, we performed experiments to exclude a GstI-mediated mechanism of glutamine synthetase II inhibition/degradation. Finally, the protein was over expressed in Escherichia coli, purified and characterised. PMID- 14759515 TI - Altered channel gating mechanism for CFTR inhibition by a high-affinity thiazolidinone blocker. AB - The thiazolidinone CFTR(inh)-172 was identified recently as a potent and selective blocker of the cystic fibrosis transmembrane conductance regulator (CFTR) Cl(-) channel. Here, we characterized the CFTR(inh)-172 inhibition mechanism by patch-clamp and short-circuit analysis using cells stably expressing wild-type and mutant CFTRs. CFTR(inh)-172 did not alter CFTR unitary conductance (8 pS), but reduced open probability by >90% with K(i) approximately 0.6 microM. This effect was due to increased mean channel closed time without changing mean channel open time. Short-circuit current experiments indicated similar CFTR(inh) 172 inhibitory potency (K(i) approximately 0.5 microM) for inhibition of Cl(-) current in wild-type, G551D, and G1349D CFTR; however, K(i) was significantly reduced to 0.2 microM for DeltaF508 CFTR. Our studies provide evidence for CFTR inhibition by CFTR(inh)-172 by a mechanism involving altered CFTR gating. PMID- 14759516 TI - Interaction of checkpoint kinase 1 and the X-linked inhibitor of apoptosis during mitosis. AB - We report here that the checkpoint kinase Chk1 and the inhibitor of apoptosis protein (IAP) family member XIAP can be found in a complex in association with condensed chromosomes aligned at the metaphase plate during mitosis. The interaction between Chk1 and XIAP was transient and followed the breakdown of the nuclear envelope. Chk1 and XIAP also formed a complex in vitro and in coimmunoprecipitation experiments. The interaction between Chk1 and the BIR3 domain of XIAP in vitro required an N-terminal sequence in Chk1 that is identical to the BIR-binding motif at the N-terminus of HID. An interaction of Chk1 and XIAP may imply a mechanism of coupling between the regulatory networks that control cell cycle progression and apoptosis during mitosis. PMID- 14759517 TI - Conjugate for efficient delivery of short interfering RNA (siRNA) into mammalian cells. AB - The efficient delivery of short interfering RNAs (siRNAs) into cells provides a powerful approach to study cellular functions. SiRNAs were coupled to the membrane permeant peptides (MPPs) penetratin and transportan to improve their uptake by cells. Thiol-containing siRNAs corresponding to luciferase, or green fluorescent protein (GFP) transgenes, were synthesized and conjugated to penetratin or transportan via a disulfide bond that is labile in the reducing environment of the cytoplasm. These MPP-siRNAs efficiently reduced transient and stable expression of reporter transgenes in several mammalian cell types in a high proportion of cells, and demonstrated equivalent or better delivery characteristics than cationic liposomes with fewer manipulations. PMID- 14759518 TI - Functional silencing of hepatic microsomal glucose-6-phosphatase gene expression in vivo by adenovirus-mediated delivery of short hairpin RNA. AB - An expression cassette containing mouse U6 polymerase III promoter directing expression of short hairpin RNA (shRNA) targeting murine microsomal glucose-6 phosphatase (G6P) transcript was generated. This construct was packaged into an adenoviral (AdV) backbone and viral stocks generated. Mice injected intravenously with AdV-G6PshRNA exhibited a significant reduction in postprandial glucose levels and had significantly elevated steady-state hepatic glycogen stores. Target gene silencing was confirmed by measurements demonstrating a significant reduction in both hepatic G6P transcript level and phosphohydrolase activity. These findings provide evidence that AdV delivery of expressed shRNA can be a productive tool to explore gene function in vivo. PMID- 14759519 TI - The peptide methionine sulfoxide reductases, MsrA and MsrB (hCBS-1), are downregulated during replicative senescence of human WI-38 fibroblasts. AB - In contrast to other oxidative modifications of amino acids, methionine sulfoxide can be enzymatically reduced back to methionine in proteins by the peptide methionine sulfoxide reductase system, composed of MsrA and MsrB. The expression of MsrA and one member of the MsrB family, hCBS-1, was analyzed during replicative senescence of WI-38 human fibroblasts. Gene expression decreased for both enzymes in senescent cells compared to young cells, and this decline was associated with an alteration in catalytic activity and the accumulation of oxidized proteins during senescence. These results suggest that downregulation of MsrA and hCBS-1 can alter the ability of senescent cells to cope with oxidative stress, hence contributing to the age-related accumulation of oxidative damage. PMID- 14759520 TI - Robust global sensitivity in multiple enzyme cascade system explains how the downstream cascade structure may remain unaffected by cross-talk. AB - A steady-state framework was applied to the ubiquitous tricyclic enzyme cascade structure, as seen in the mitogen-activated protein (MAP) kinase system, to analyze the effect of upstream kinase concentrations on final output response. The results suggest that signal amplification achieved by the cascade structure ensured that the modifying enzymes at various steps of the cascade were nearly saturated. Thus, there was no change in the response sensitivity with increasing upstream kinase concentration. Analysis was also extended to branching of a signaling pathway as an example of cross-talk. It was observed that the cascade structure confers a larger share of the signal transduction properties to its last kinase. This phenomenon in enzyme cascades may explain how the response of the terminal MAP kinase is unaffected by cross-talk of upstream kinases. PMID- 14759521 TI - Histidine-containing phosphotransfer domain extinction by RNA interference turns off a cytokinin signalling circuitry in Catharanthus roseus suspension cells. AB - We previously reported that cytokinins (CK) induce the fast and specific transcription of CrRR1, a gene encoding a type A response regulator in Catharanthus roseus cell cultures. Here, we characterized the CrHPt1 gene that encodes a histidine-containing phosphotransfer domain. CrHPt1 was silenced through RNA interference (RNAi) to test its possible implication in the CK signalling pathway. In transgenic lines stably transformed with an intron-spliced construct, the degradation of CrHPt1 transcripts abolishes the CK inductive effect on CrRR1 transcription. These result give a new in vivo functional argument for the crucial role of HPt proteins in the CK signalling pathway leading to the expression of the genes encoding type A response regulators. They also show that RNAi is a powerful strategy to turn off the CK signalling circuitry. PMID- 14759522 TI - Long-lasting RNAi activity in mammalian neurons. AB - The effect of RNA interference (RNAi) induced by synthetic small interfering RNAs (siRNAs) on proliferating mammalian cells appears to last for approximately 3-7 days after its induction. Here we show that the RNAi activity induced by a synthetic 21-nucleotide siRNA duplex in postmitotic neurons, mouse primary hippocampal neurons and neurons that differentiated from mouse embryonal carcinoma P19 cells persists for at least 3 weeks, suggesting long-lasting RNAi activity in mammalian neurons. In addition, we also show that an apoptotic (or antiviral) pathway triggered by long dsRNAs is generated during neuronal differentiation of P19 cells, by which the sequence-specific RNAi activity involving long dsRNA appears to be masked. PMID- 14759523 TI - Aryl hydrocarbon receptor-mediated suppression of GH receptor and Janus kinase 2 expression in mice. AB - Differential mRNA display revealed that a cDNA encoding the major urinary protein 2 (MUP2) that belongs to the lipocalin superfamily was absent in livers of mice treated with 3-methylcholanthrene (MC). The expression of MUP2 is known to be stimulated by growth hormone (GH), through the GH receptor (GHR), Janus kinase 2 (JAK2) and signal transducer and activator of transcription 5 (STAT5) signal transduction pathway. Since MC is an aryl hydrocarbon receptor (AhR) ligand, the effects of MC treatment on the expression of GHR, JAK2 or STAT5 in the livers of wild-type or AhR-null mice were examined. The result indicated that the expression of GHR and JAK2 mRNA was greatly decreased by MC in wild-type mice but not in AhR-null mice. In addition, the binding activity of STAT5 bound to STAT5 binding element was reduced after MC treatment in wild-type mice but not in AhR null mice. Based on these results, we conclude that the suppression of MUP2 mRNA expression by MC is caused by the AhR-mediated disruption of the GH signaling pathway. Possible mechanism(s) by which exposure to aromatic hydrocarbons causes a decrease in the body weight of mice, which has been referred to as wasting syndrome, will also be discussed. PMID- 14759524 TI - The solution structure of the SODD BAG domain reveals additional electrostatic interactions in the HSP70 complexes of SODD subfamily BAG domains. AB - The solution structure of an N-terminally extended construct of the SODD BAG domain was determined by nuclear magnetic resonance spectroscopy. A homology model of the SODD-BAG/HSP70 complex reveals additional possible interactions that are specific for the SODD subfamily of BAG domains while the overall geometry of the complex remains the same. Relaxation rate measurements show that amino acids N358-S375 of SODD which were previously assigned to its BAG domain are not structured in our construct. The SODD BAG domain is thus indeed smaller than the homologous domain in Bag1 defining a new subfamily of BAG domains. PMID- 14759525 TI - Integrin beta1 subunit overexpressed in the SMMC-7721 cells regulates the promoter activity of p21(CIP1) and enhances its transcription. AB - Evidence has been emerging to suggest that integrin could induce growth inhibition in some cell types. Some of the molecular mechanisms underlying growth arrest have been elucidated. We reported here that overexpression of integrin beta1 imposed a growth inhibitory effect on the hepatocellular carcinoma cell line SMMC-7721, and this phenomenon was mainly attributed to the cyclin-dependent kinase inhibitor p21(CIP1). Furthermore, our findings suggested that transcription activity of the p21(CIP1) gene could be upregulated in the integrin beta1-overexpressing cells, and possibly controlled by the cis-elements in the core region of the p21(CIP1) promoter. PMID- 14759526 TI - Identification of binding domains in the sodium channel Na(V)1.8 intracellular N terminal region and annexin II light chain p11. AB - The interaction of p11 (annexin II light chain) with the N-terminal domain of Na(V)1.8, a tetrodotoxin-resistant sodium channel, is essential for the functional expression of the channel. Here we show that p11 binds to Na(V)1.8 but not to sodium channel isoforms Na(V)1.2, 1.5, 1.7 or Na(V)1.9. The binding of amino acids 74-103 of Na(V)1.8 to p11 residues 33-78 occurs in a random coiled region flanked by two EF hand motifs whose crystal structure has been established. As Na(V)1.8 channel expression is associated with pain pathways, drugs that disrupt the Na(V)1.8-p11 interaction and down-regulate channel expression may have analgesic activity. PMID- 14759527 TI - Quivisianthone, an evodulone limonoid from the Madagascan Meliaceae Quivisia papinae. AB - An investigation of the seeds of the Madagascan Meliaceae Quivisia papinae has yielded quivisianthone, a novel evodulone group limonoid, together with the known azadiradione and two novel derivatives: 6 alpha-hydroxyazadiradione and 7 deacetyl-7-angeloyl-6 alpha-hydroxyazadiradione. Quivisianthone is the first reported evodulone group limonoid possessing both a ring A lactone and an azadiradione-type ring D. PMID- 14759528 TI - Four illudane sesquiterpenes from Coprinopsis episcopalis. AB - Four new illudane derivatives with antibiotic and cytotoxic properties, illudins I (1), I(2) (2), J (3) and J(2) (4), have been isolated from the fungus Coprinopsis episcopalis (syn. Coprinus episcopalis). These sesquiterpenes are stereoisomers, and their relative structures have been determined taking into consideration 2D NMR data. PMID- 14759529 TI - Neo-clerodane diterpenes from Teucrium fruticans. AB - In addition to three out of the four previously reported neo-clerodanes already found in Teucrium fruticans (fruticolone, isofruticolone and 8 beta hydroxyfruticolone), and 6-acetylteucjaponin B (isolated from T. scordium and T. grisebachii), four new neo-clerodanes, namely 7 beta-hydroxyfruticolone, 11 hydroxyfruticolone, deacetylfruticolone and 6-acetyl-10-hydroxyteucjaponin B were also isolated. The structures were unambiguously elucidated based on extensive NMR spectral studies (one- and two-dimensional experiments). Semi-prep-HPLC proved to be a convenient purification procedure, occasionally being followed by TLC. The new compounds were assayed against Spodoptera littoralis and two of them were shown to have potent antifeedant activity. PMID- 14759530 TI - Lanostanes and friedolanostanes from the bark of Garcinia speciosa. AB - The CHCl(3) extract of the bark of Garcinia speciosa contained four 17,14 friedolanostanes and five lanostanes as well as friedelin and common plant constituents. The friedolanostanes were the previously known methyl ester of (24E)-3 alpha,23 alpha-dihydroxy-17,14-friedolanostan-8,14,24-trien-26-oic acid and the methyl esters of three hitherto unknown acids, 3 alpha-hydroxy-16 alpha,23 alpha-epoxy-17,14-friedolanostan-8,14,24-trien-26-oic acid, 3 alpha,23 alpha-dihydroxy-8 alpha,9 alpha-epoxy-17,14-friedolanostan-15-oxo-24-en-26-oic acid and 3 alpha,23 alpha-dihydroxy-17,14-friedolanostan-15-oxo-8(14),24-dien-26 oic acid. New lanostanes were 3 beta,9 alpha-dihydroxylanost-24-en-26-al and the methyl ester of 3 beta-hydroxy-23-oxo-9,16-lanostadien-26-oic acid. Structures were established by analysis of spectroscopic data. In the case of the lanostanes the previously unassigned C-25 stereochemistry was shown to be 25R by X-ray analysis of 3 beta-hydroxy-23-oxo-9,16-lanostadien-26-oic acid. In the case of the friedolanostanes the configuration at C-23 was established as 23R, identical with the absolute configuration at C-23 of mariesiic acids A and B. PMID- 14759531 TI - (+/-)-Schefflone: a trimeric monoterpenoid from the root bark of Uvaria scheffleri. AB - The trimeric monoterpene and mildly mosquito larvicidal agent, (+/-)-schefflone, that is an apparent derivative of the antiparasitic aromatic monoterpene espintanol, was isolated from the antimalarial extracts of the root bark of Uvaria scheffleri, together with espintanol. Structural determination of (+/-) schefflone was achieved from spectroscopic data and confirmed by single-crystal X ray diffraction analysis. (+/-)-Schefflone can be considered a product of a non enzymatic Diels-Alder-type cycloaddition reaction of the quinonemethide derivative of espintanol as the diene and dienophile. PMID- 14759532 TI - Anthocyanin from strawberry (Fragaria ananassa) with the novel aglycone, 5 carboxypyranopelargonidin. AB - An anthocyanin, 1, with the novel 4-substituted aglycone, 5 carboxypyranopelargonidin, was isolated in small amounts from the acidified, methanolic extract of strawberries, Fragaria ananassa Duch., by preparative HPLC after purification by partition against ethyl acetate, Amberlite XAD-7 and Sephadex LH-20 column chromatography. It was identified mainly by 2D NMR spectroscopy and electrospray LC-MS as the 3-O-beta-glucopyranoside of 5-carboxy 2-(4-hydroxyphenyl)-3,8-dihydroxy-pyrano[4,3,2-de]-1-benzopyrylium, an anthocyanidin which is homologous to 5-carboxypyranomalvidin (vitisidin A) reported in red wines and 5-carboxypyranocyanidin recently isolated from red onions. By comparison of UV-Vis absorption spectra, 1 showed in contrast to 2, pelargonidin 3-O-beta-glucopyranoside, a local absorption peak around 360 nm, a hypsochromic shift (8 nm) of the visible absorption maximum, and lack of a distinct UV absorption peak around 280 nm. The similarities between the absorption spectra of 1 in various acidic and neutral buffer solutions implied restricted formation of the instable colourless equilibrium forms, which are typical for most anthocyanins in weakly acidic solutions. The molar absorptivity (epsilon) of 1 varied little with pH contrary to similar values of for instance the major anthocyanin in strawberry, 2. However, 2 revealed higher epsilon-values than 1 at all pH values except 5.1. At pH 5.1, the epsilon-value of 1 (6250) was nearly four times the corresponding value of 2 (1720), which showed the potential of 5-carboxypyranopelargonidin derivatives as colorants in solutions with pH around 5. The colours of 1 and 2 in buffered solutions with pH 1.1 and pH 6.9 have been described by the CIELAB coordinates h(ab) (hue angle), C* (chroma), and L* (lightness). PMID- 14759533 TI - Detection of hypericins in the "red glands" of Hypericum elodes by ESI-MS/MS. AB - The biologically active naphthodianthrones hypericin and pseudohypericin were detected by electrospray ionization mass spectrometry (ESI-MS/MS) in microsamples from the sepals of Hypericum elodes (Hypericaceae) containing the so-called "red glands", i.e. stipitate glands with red-coloured heads. The occurrence of hypericins in the red glands of H. elodes supports the taxonomic position of the section Elodes within the genus Hypericum and provides evidence that the ability of carrying out the biosynthetic pathway leading to the naphthodianthrone compounds, rather than the absolute amounts produced, should be regarded as a chemical marker of the phylogenetically more advanced sections of genus Hypericum. The biologically active phloroglucinol derivatives hyperforin and adhyperforin, so far found only in H. perforatum, were also detected and evidence for their localization in the sepal secretory canals with large lumen, is given. PMID- 14759534 TI - Structural investigations on betacyanin pigments by LC NMR and 2D NMR spectroscopy. AB - Four betacyanin pigments were analysed by LC NMR and subjected to extensive NMR characterisation after isolation. Previously, low pH values were applied for NMR investigations of betalains resulting in rapid degradation of the purified substances thus preventing extensive NMR studies. Consequently, up to now only one single (13)C NMR spectrum of a betalain pigment, namely that of neobetanin (=14,15-dehydrobetanin), was available. Because of its sufficient stability under highly acidic conditions otherwise detrimental for betacyanins, this pigment remained an exemption. Since betalains are most stable in the pH range of 5-7, a new solvent system has been developed allowing improved data acquisition through improved pigment stability at near neutral pH. Thus, not only (1)H, but for the first time also partial (13)C data of betanin, isobetanin, phyllocactin and hylocerenin isolated from red-purple pitaya [Hylocereus polyrhizus (Weber) Britton & Rose, Cactaceae] could be indirectly obtained by gHSQC- and gHMQC-NMR experiments. PMID- 14759535 TI - L-O-Caffeoylhomoserine from Matteuccia struthiopteris. AB - A caffeic acid derivative was isolated from Matteuccia struthiopteris (ostrich fern) as a major radical scavenger. The compound consisted of caffeic acid and L homoserine. NMR and MS analysis revealed the structure as L-O-caffeoylhomoserine. PMID- 14759536 TI - Diprenylated chalcones and other constituents from the twigs of Dorstenia barteri var. subtriangularis. AB - The twigs of Dorstenia barteri var. subtriangularis yielded three diprenylated chalcones: (-)-3-(3,3-dimethylallyl)-5'-(2-hydroxy-3-methylbut-3-enyl)-4,2',4' trihydroxychalcone, (+)-3-(3,3-dimethylallyl)-4',5'-[2'''-(1-hydroxy-1 methylethyl)-dihydrofurano]-4,2'-dihydroxychalcone and 3,4-(6",6" dimethyldihydropyrano)-4',5'-[2''',-(1-hydroxy-1-methylethyl)-dihydrofurano]-2' hydroxychalcone for which the names bartericins A, B and C, respectively, are proposed. Stipulin, beta-sitosterol and its 3-beta-D-glucopyranosyl derivative were also isolated. The structures of these secondary metabolites were determined on the basis of spectroscopic analysis, especially, NMR spectra in conjunction with 2D experiments, COSY, HMQC and HMBC. The structural relationship of bartericins B and C was further established by the chemical cyclization of one to the other. PMID- 14759537 TI - Cyclobutanes from Combretum albopunctatum. AB - A dichloromethane extract of the aerial parts of Combretum albopunctatum Suesseng afforded five phenolic compounds-three known flavonoids and two novel cyclobutane chalcone dimers. The chemical structures were determined by standard spectroscopic techniques and the structure and relative stereochemistry of one chalcone dimer, rel-(1 alpha,2 beta)-di-(2,6-dimethoxy-4-hydroxy)-benzoyl-rel-(3 alpha,4 beta)-diphenylcyclobutane, were confirmed by single crystal X-ray diffraction. PMID- 14759538 TI - Furanoflavonoids from Pongamia pinnata fruits. AB - Fruits of Pongamia pinnata afforded four new furanoflavonoids, pongapinnol A-D (1 4), and a new coumestan, pongacoumestan (5) along with thirteen known compounds 6 18. Compounds 16 and 17 are isolated for the first time from this plant. The structures of isolated compounds were elucidated on the basis of spectroscopic data interpretation. PMID- 14759539 TI - Flavonoids from Cleistocalyx operculatus. AB - Two flavonoids 3'-formyl-4',6'-dihydroxy-2'-methoxy-5'-methylchalcone and (2S)-8 formyl-5-hydroxy-7-methoxy-6-methylflavanone together with five known compounds, were isolated from the dried buds of Cleistocalyx operculatus. Their structures were determined on the basis of spectroscopic analyses (UV, IR, EIMS, (1)H, (13)C NMR and HMBC). PMID- 14759540 TI - N,beta-D-Glucopyranosyl vincosamide, a light regulated indole alkaloid from the shoots of Psychotria leiocarpa. AB - From leaves of Psychotria leiocarpa, an indole alkaloid was isolated to which the structure N,beta-D-glucopyranosyl vincosamide (1) was assigned. This represents the first report of an N-glycosylated monoterpenoid indole alkaloid. In field grown plants highest amounts of 1 were found in the leaves (2.5% of dry wt) and fruit pulp (1.5% dry wt). Lower amounts were found in the stems (0.2% dry wt) and the seeds (0.1% of dry wt), whereas the alkaloid was not detected in the roots. The accumulation of 1 in aseptic seedlings was also restricted to the shoots and increased with plant age and light exposure, independent of the supply of sucrose in the culture medium. PMID- 14759541 TI - Enteridinines A and B from slime mold Enteridium lycoperdon. AB - Two novel deoxysugar esters, named enteridinines A and B, were isolated from the slime mold Enteridium lycoperdon. Their structures, including the absolute configurations of the hydroxyl and methyl groups, were determined by means of extensive spectroscopic data such as UV, IR, MS, 1D and 2D NMR spectra. Enteridinines A and B have unique structures containing 1,7 dioxaspiro[5.5]undecanes with an O-beta-D-mycarosyl-(1-->4)-beta-D-olivosyl and an O-beta-L-olivomycosyl-(1-->4)-beta-D-amicetosyl-(1-->4)-beta-L-digitoxosyl unit, respectively, and showed growth inhibitory activities against Gram positive bacteria. PMID- 14759542 TI - C,O-bisglycosylapigenins from the leaves of Rhamnella inaequilatera. AB - From the leaves of Rhamnella inaequilatera, three flavone C,O-bisglycosides, rhamnellaflavosides A, B and C, were isolated and their structures were elucidated based on their spectral data and chemical evidence. PMID- 14759543 TI - Cyclic azaphilones daldinins E and F from the ascomycete fungus Hypoxylon fuscum (Xylariaceae). AB - Phytochemical examination of the methanolic extract of fruit bodies of the Xylariaceous ascomycete fungus Hypoxylon fuscum led to the isolation of two azaphilone derivatives named daldinins E and F together with two known compounds daldinin C and 4,5,4',5'-tetrahydroxy-1:1'-binaphthyl using a combination of reversed phase HPLC and high performance gel permeation chromatography. Their structures were determined by 2D NMR, MS, IR, UV, and CD spectroscopy. Their antioxidative activities were also estimated by an indication of a 1,1-diphenyl-2 picrylhydrazyl (DPPH) radical-scavenging effect. The chemosystematics of Hypoxylon is discussed. PMID- 14759544 TI - Ascosonchine, the enol tautomer of 4-pyridylpyruvic acid with herbicidal activity produced by Ascochyta sonchi. AB - A new phytotoxic enol tautomer of 4-pyridylpyruvic acid, named ascosonchine, was isolated from the culture filtrate of Ascochyta sonchi. Such a leaf pathogen is a potential biocontrol agent of Sonchus arvensis, a perennial herbaceous weed occurring throughout the temperate regions of the world. Ascosonchine, characterised as (Z)-2-hydroxy-3-(4-pyridyl)-2-propenoic acid by spectroscopic methods, showed selective herbicidal properties, that are not associated with antibacterial, antifungal or zootoxic activities. PMID- 14759545 TI - Maltol glucosides from the tuber of Smilax bockii. AB - Two maltol glucosides, bockiosides A and B, along with 10 known compounds, were isolated from the tuber of Smilax bockii (Liliaceae), and their structures were elucidated by spectral experiments, chemical analysis and comparison with literature data. PMID- 14759546 TI - Lasianthionosides A-C, megastigmane glucosides from leaves of Lasianthus fordii. AB - From the leaves of Lasianthus fordii, three megastigmane glucosides, lasianthionosides A, B and C, were isolated together with the known iridoid glucoside, asperuloside, deacetylasperuloside and methyl deacetyl-asperulosidate, and a megastigmane glucoside, citroside A. The structures have been elucidated based on spectroscopic analyses and/or X-ray crystallographic analysis. PMID- 14759547 TI - Fascicularones A and B from a mycelial culture of Naematoloma fasciculare. AB - Two sesquiterpenoids, fascicularones A and B, have been isolated from the culture broth of a poisonous mushroom, Naematoloma fasciculare. Their structures were determined using spectroscopic methods. PMID- 14759548 TI - Structure elucidation and phytotoxicity of C13 nor-isoprenoids from Cestrum parqui. AB - Twelve C(13) nor-isoprenoids have been isolated from the leaves of Cestrum parqui (Solanaceae). The structure (2R,6R,9R)-2,9-dihydroxy-4-megastigmen-3-one has been assigned to the new compound. All the structures have been determined by spectroscopic means and chemical correlations. The compounds showed phytotoxic effect on the germination and growth of Lactuca sativa L. PMID- 14759549 TI - Cytotoxic cardenolide glycoside from the seeds of Cerbera odollam. AB - A cardenolide glycoside, 3 beta-O-(2'-O-acetyl-l- thevetosyl)-15(14-->8)-abeo-5 beta-(8R)-14-oxo-card-20(22)-enolide (2'-O-acetyl cerleaside A), was isolated from a methylene chloride extract of the seeds of Cerbera odollam, together with four known compounds: cerleaside A, 17 alpha-neriifolin, 17 beta- neriifolin and cerberin. Their structures were elucidated by spectroscopic methods. All compounds except cerleaside A exhibited cytotoxic activities against oral human epidermoid carcinoma (KB), human breast cancer cell (BC) and human small cell lung cancer (NCI-H187). PMID- 14759550 TI - Changes in mechanism of PACAP-induced relaxation in longitudinal muscle of the distal colon of Wistar rats with age. AB - Mechanisms of relaxation of longitudinal muscle of the distal colon induced by exogenously added pituitary adenylate cyclase activating peptide (PACAP) were studied in 2- to 30-week-old Wistar rats. Exogenous PACAP induced very significant relaxation of the longitudinal muscle in 2-week-old rats, but this effect decreased significantly with age. The cyclic AMP-cyclic AMP-dependent protein kinase (PKA) pathway and the tyrosine kinase-small conductance Ca2+ activated K+ channel (SK channel) pathway were found to be involved in the mechanism of PACAP-induced relaxation. In 2-week-old rats, PACAP-induced relaxation was significantly inhibited by tetrodotoxin (TTX). Since relaxation was also significantly inhibited by NG-nitro-L-arginine (N5-nitro-amidino-L-2,5 diamino-pentanoic acid: L-NOARG), the neurogenic effect of PACAP seems to be mediated mainly through nitric oxide neurons. In 8-week-old rats, L-NOARG and TTX had little effect on PACAP-induced relaxation, suggesting that the relaxant effect in 8-week-old rats is a direct action on longitudinal smooth muscle cells. Changes in the mechanisms of PACAP-induced relaxation with age were examined in the distal colon in relation to changes in the neurogenic and the direct effects of PACAP. The neurogenic effect in the exogenous PACAP-induced relaxation of the longitudinal muscle of the Wistar rat distal colon is dominant in tissue isolated from 2-week-old and lost in tissue isolated from 8-week-old rats. PMID- 14759551 TI - Glucagon-like peptide-1 secretion is influenced by perfusate glucose concentration and by a feedback mechanism involving somatostatin in isolated perfused porcine ileum. AB - Glucagon-like peptide-1 (GLP-1) is released from intestinal L-cells in response to ingestion of meals. The mechanisms regulating its secretion are not clear, but local somatostatin (SS) restrains GLP-1 secretion. We investigated feedback and substrate regulation of GLP-1 and SS secretion, using isolated perfused porcine ileum (n=17). Effluents were measured for GLP-1 and SS. Perfusion pressure and motility were recorded. Investigated parameters included spontaneous fluctuations, changes in perfusate glucose concentrations (3.5, 5, 11 mM) and addition of insulin (1 nM). We also investigated the effect of proglucagon products, glucagon (10 nM), GLP-1 and GLP-2 (0.1, 1, and 10 nM) on GLP-1 and SS secretion, as well as on glucagon-like peptide-2 (GLP-2), peptide YY (PYY) and GIP secretion, all possible product of L-cells or neighbour cells. Perfusate glucose concentration dose-dependently stimulated GLP-1 secretion (p=0.011). Insulin had no effect. Glucagon weakly stimulated GIP secretion. GLP-1 stimulated SS secretion and motor activity, but inhibited GLP-2, GIP and PYY secretion and perfusion pressure. GLP-2 weakly stimulated SS secretion. We conclude (a) that GLP-1 secretion is influenced by perfusate glucose concentration and (b) that L cell secretion is feedback regulated by GLP-1 itself, probably via paracrine SS activity. PMID- 14759552 TI - BDNF-overexpression regulates the reactivity of small pulmonary arteries to neurokinin A. AB - Brain-derived neurotrophic factor (BDNF) is a key modulator during the development of jugular and nodose ganglia neurons, which represent the origin of a large proportion of the sensory innervation of the lung. It belongs to the family of neurotrophins, which have been shown to induce the expression of tachykinins. To assess the interactions of BDNF and the tachykinin neurokinin A (NKA) in small pulmonary vessels, BDNF-transgenic mice were examined for tachykinin contents in the airways, heart, trigeminal ganglion and jugular and nodose ganglion complex (JNC) using reverse phase HPLC (rpHPLC) and radioimmunoassay. BDNF-overexpression led to increased NKA levels in the heart and the JNC, whereas only slightly enhanced levels in the trigeminal ganglion were detected. Lower NKA levels were found in the lung. To assess vasoreactivity in small arteries, precision cut lung slices were subjected to videomorphometry and the response to NKA was examined, which showed significantly stronger effects in the BDNF-transgenic mice, while NK-2 receptor mRNA expression, assayed by real time RT-PCR, was reduced. In conclusion, BDNF-overexpression results in decreased levels of NKA in the lung with subsequently increased NKA-sensitivity of small arteries. These findings point to a modulatory role of neurotrophins in small respiratory vessel tone regulation. PMID- 14759553 TI - Functional analysis of the SGNP I in the pupal diapause of the oriental tobacco budworm, Helicoverpa assulta (Lepidoptera: Noctuidae). AB - Helicoverpa assulta suboesophageal ganglion neuropeptide I (Has-SGNP I) is a 24 amino acids peptide amide, which shows 62.5% similarity with the diapause hormone of Bombyx mori (Bom-DH). It has been demonstrated that embryonic diapause is induced by DH in B. mori. Injection of synthetic amidated Has-SGNP I terminated pupal diapause in a dose-dependent manner. Therefore, Has-SGNP I might be referred to a "diapause termination hormone" in H. assulta (Has-DTH). The maximal dose of Has-DTH for diapause termination was 1.0 microg and the half-maximal dose 0.4 microg. The time required for diapause termination of Has-DTH was 2-3 days longer than that of 20-hydroxyecdysone. During the pupal stage, DTH mRNA content in the SGs of nondiapausing pupae was always higher than in diapausing pupae using the combined method of quantitative RT-PCR and Southern blot. DTH gene also expressed at a low level while diapausing pupae were chilled at 4 degrees C, but increased rapidly and largely after being transferred to 25 degrees C. Using a competitive ELISA, Has-DTH-like immunoreactivity in the haemolymph showed the same pattern as that of Has-DTH gene expression. Those results indicated that Has DTH gene expression was related to diapause development and could be activated by low temperature. Has-DTH might be useful to elucidate the mechanism of diapause termination in pupal diapause species. PMID- 14759554 TI - Effects of intracerebroventricularly injected glucagon-like peptide-1 on cardiovascular parameters; role of central cholinergic system and vasopressin. AB - We aimed to investigate the effects of intracerebroventricularly (i.c.v.) injected glucagon-like peptide-1 (GLP-1) on blood pressure and heart rate, and whether central cholinergic system and vasopressinergic system play roles in these effects. Male Wistar albino rats were used throughout the experiments. Blood pressures and heart rates were observed before and for 30 min following drug injections. i.c.v. GLP-1 (100, 500 and 1000 ng/10 microl) caused a dose dependent increase in both blood pressure and heart rate. Nicotinic receptor antagonist mecamylamine (25 microg/10 microl, i.c.v.) and muscarinic receptor antagonist atropine (5 microg/10 microl, i.c.v.) prevented the stimulating effect of GLP-1 on blood pressure. The effect of GLP-1 on heart rate was blocked only by mecamylamine. The V1 receptor antagonist of vasopressin (B-mercapto B, B cyclopentamethylenepropionyl, O-Me-Tyr,Arg)-vasopressin (10 microg/kg), that was applied intraarterially, only prevented the effect of GLP-1 on blood pressure, but did not show any effect on heart rate. Our data indicate that i.c.v. GLP-1 increases blood pressure and heart rate, and stimulation of central nicotinic and partially muscarinic receptors and vasopressinergic system play a role in the effects of i.c.v. GLP-1 on blood pressure. The effect of GLP-1 on heart rate may be partially due to stimulation of central nicotinic receptors. PMID- 14759555 TI - Corticotropin-releasing hormone (CRH) produces analgesia in a thermal injury model independent of its effect on systemic beta-endorphin and corticosterone. AB - To determine separately the effect of corticotropin-releasing hormone (CRH) on analgesia and on inflammation, rats were assigned to receive CRH 60 microg/kg, CRH 300 microg/kg, morphine 4 mg/kg, or normal saline intravenously 15 min before a burn injury. Two mesh chambers that allowed collection of fluid had been previously implanted subdermally in each rat. The skin overlying the right chamber was subject to thermal injury. The left chamber served as a control. We assessed systemic analgesia, and levels of beta-endorphin and corticosterone in plasma and in chamber fluid before, 1, 4 and 24 h after drug administration. The CRH groups exhibited longer tail flick latencies than the control group (P=0.0001) although the increase in latency was of smaller magnitude than in the morphine group. We did not observe a CRH dose response for analgesia. Plasma corticosterone levels were higher in the CRH 300 microg/kg group than in the normal saline group at 4 h (P=0.03). Levels of beta-endorphin in plasma as well as the levels of corticosterone and beta-endorphin in chambers were similar in the CRH 300 microg/kg group and in the normal saline group (all P values>0.1). Thus, systemically administered CRH produces analgesia in thermal injury independent of its effect on these two markers of local or systemic inflammation. PMID- 14759556 TI - Angiotensin-(1-7) inhibits the angiotensin II-enhanced norepinephrine release in coarcted hypertensive rats. AB - Since it has been suggested that angiotensin (Ang) (1-7) functions as an antihypertensive peptide, we studied its effect on the Ang II-enhanced norepinephrine (NE) release evoked by K+ in hypothalami isolated from aortic coarcted hypertensive (CH) rats. The endogenous NE stores were labeled by incubation of the tissues with 3H-NE during 30 min, and after 90 min of washing, they were incubated in Krebs solution containing 25 mM KCl in the absence or presence of the peptides. Ang-(1-7) not only diminished the K+-evoked NE release from hypothalami of CH rats, but also blocked the Ang II-enhanced NE release induced by K+. Ang-(1-7) blocking action on the Ang II response was prevented by [D-Ala7]Ang-(1-7), an Ang-(1-7) specific antagonist, by PD 123319, an AT2 receptor antagonist, and by Hoe 140, a B2 receptor antagonist. Ang-(1-7) inhibitory effect on the Ang II facilitatory effect on K+-stimulated NE release disappeared in the presence of Nomega-nitro-L-arginine methylester and was restored by L-arginine. Our present results suggest that Ang-(1-7) may contribute to blood pressure regulation by blocking Ang II actions on NE release at the central level. This inhibitory effect is a nitric oxide-mediated mechanism involving AT2 receptors and/or Ang-(1-7) specific receptors and local bradykinin generation. PMID- 14759557 TI - Modulatory effect of endothelin-1 and -3 on neuronal norepinephrine release in the rat posterior hypothalamus. AB - Based upon the existence of high density of ET-receptors on catecholaminergic neurons of the hypothalamus, we studied the effects of endothelin-1 (ET-1) and endothelin-3 (ET-3) on neuronal norepinephrine (NE) release in the rat posterior hypothalamus. The intracellular pathways and receptors involved were also investigated. Neuronal NE release was enhanced by ET-1 and ET-3 (10 etaM). The selective antagonists of subtype A and B ET receptors (ETA, ETB) (100 etaM BQ-610 and 100 etaM BQ-788, respectively) abolished the increase induced by ET-1 but not by ET-3. The PLC inhibitor, U73122 (10 microM), abolished ET-1 and ET-3 response. GF-109203X (100 etaM) (PKC inhibitor) blocked the increase in NE release produced by ET-3 and partially blocked ET-1 response. The inositol 1,4,5-trisphosphate induced calcium release inhibitor, 42 microM 2-APB, inhibited the stimulatory effect induced by ET-3 but not by ET-1. The PKA inhibitor, 500 etaM H-89, blocked the increase in neuronal NE release evoked by ET-1 but not by ET-3. Our results showed that ET-1 as well as ET-3 displayed an excitatory neuromodulatory effect on neuronal NE release in the rat posterior hypothalamus. ET-1 through an atypical ETA or ETB receptor activated the PLC/PKC signalling pathway as well as the cAMP pathway, whereas ET-3 through a non-ETA/non-ETB receptor activated the phosphoinositide pathway. Both ETs would enhance the sympathoexcitatory response elicited by the posterior hypothalamus and thus participate in cardiovascular regulation. PMID- 14759558 TI - Histamine and histidine decarboxylase are hallmark features of ECL cells but not G cells in rat stomach. AB - The oxyntic mucosa of the rat stomach is rich in ECL cells which produce and secrete histamine in response to gastrin. Histamine and the histamine-forming enzyme histidine decarboxylase (HDC) have been claimed to occur also in the gastrin-secreting G cells in the antrum. In the present study, we used a panel of five HDC antisera and one histamine antiserum to investigate whether histamine and HDC are exclusive to the ECL cells. By immunocytochemistry, we could show that the ECL cells were stained with the histamine antiserum and all five HDC antisera. The G cells, however, were not stained with the histamine antiserum, but with three of the five HDC antisera. Thus, histamine and HDC coexist in the ECL cells (oxyntic mucosa) but not in G cells (antral mucosa). Western blot analysis revealed a typical pattern of HDC-immunoreactive bands (74, 63 and 54 kDa) in oxyntic mucosa extracts with all five antisera. In antral extracts, immunoreactive bands were detected with three of the five HDC antisera (same as above); the pattern of immunoreactivity differed from that in oxyntic mucosa. Food intake of fasted rats or treatment with the proton pump inhibitor omeprazole raised the HDC activity and the HDC protein content of the oxyntic mucosa but not of the antral mucosa; the HDC activity in the antrum was barely detectable. We suggest that the HDC-like immunoreactivity in the antrum represents a cross reaction with non-HDC proteins and conclude that histamine and HDC are hallmark features of ECL cells but not of G cells. PMID- 14759559 TI - Internalization of neuropeptide Y Y1 and Y5 and of pancreatic polypeptide Y4 receptors is inhibited by lithium in preference to sodium and potassium ions. AB - The receptor-linked internalization of [125I] human neuropeptide Y (NPY) in Chinese hamster ovary (CHO) cells expressing the guinea-pig Y1 receptors or in human endometrial carcinoma-1B (Hec-1B) cells expressing the human Y5 receptor, as well as the receptor-linked internalization of human pancreatic polypeptide (hPP) receptor expressed in CHO cells, is selectively inhibited by low molarities of the Li+ cation. The Na+ and K+ cations decreased the receptor-linked internalization of agonist peptides only at high molar inputs, and largely in proportion to the reduction of cell surface binding of Y ligand peptides, dependent on ion concentration and the type of Y receptor examined. With particulates isolated from disrupted cells, there was no preferential inhibition by Li+ relative to Na+ in the binding of type-specific ligand peptides to Y receptors of any type. The observed difference could be connected to the known ability of Li+ to modify active conformations of signal transducers, which may also directly or indirectly affect the internalization motors. The decrease in the rate of Y receptor internalization by Li+ also points to a possible alteration of Y receptor signaling in vivo by lithium at acute therapeutically employed dose levels. PMID- 14759560 TI - Conformational preferences and activities of peptides from the catecholamine release-inhibitory (catestatin) region of chromogranin A. AB - Previous modeling (PDB 1cfk) of the catecholamine release-inhibitory "catestatin" region of chromogranin A (CgA) suggested a beta-strand/loop/beta-strand active conformation, displaying an electropositive Arg-rich loop (R(351)AR(353)GYGFR(358)). To explore this possibility, we studied NMR structures of linear and cyclic synthetic catestatin, bovine (bCgA(344-364)) or human (hCgA(352-372)). By 2-D (1)H-NMR, the structure of linear catestatin (hCgA(352 372)) exhibited the NOE pattern of a coiled loop (PDB 1lv4). We then constrained the structure, cyclizing the putative Arg-rich loop connecting the beta-strands: cyclic bCgA(350-362) ([C(0)]F(350)RARGYGFRGPGL(362)[C(+14)]). Favored conformations of cyclic bCgA(350-362) were determined by (1)H-NMR and (13)C-NMR spectroscopy. Cyclic bCgA(350-362) conformers (PDB 1n2y) adopted a "twisted-loop" conformation. Alignment between the homology model and the cyclic NMR structure showed that, while portions of the NMR structure's mid-molecule and carboxy terminus were congruent with the homology model (RMSD, 1.61-1.91 A), the amino terminal "twisted loop" coiled inward and away from the model (RMSD, 3.36 A). Constrained cyclic bCgA(350-362) did not exert nicotinic cholinergic antagonist activity (IC(50)>10 microM), when compared to full-length linear (IC(50) approximately 0.42-0.56 microM), or cyclic (IC(50) approximately 0.74 microM) catestatin. Thus, loss of activity in the small, constrained peptide did not result from either [Cys]-extension or cyclization, per se. While linear catestatin displays coiled character, a small cyclic derivative lost biological activity, perhaps because its amino-terminal domain deviated sharply from the predicted active conformation. These results refine the relationship between structure and function in catestatin, and suggest goals in future peptidomimetic syntheses, in particular attempts to constrain the correct amino-terminal shape for biological activity. PMID- 14759561 TI - Blood glucose control in healthy subject and patients receiving intravenous glucose infusion or total parenteral nutrition using glucagon-like peptide 1. AB - AIMS: It was the aim of the study to examine whether the insulinotropic gut hormone GLP-1 is able to control or even normalise glycaemia in healthy subjects receiving intravenous glucose infusions and in severely ill patients hyperglycaemic during total parenteral nutrition. PATIENTS AND METHODS: Eight healthy subjects and nine patients were examined. The volunteers received, in six separate experiments in randomised order, intravenous glucose at doses of 0, 2 and 5mg kg(-1) min(-1), each with intravenous GLP-1 or placebo for 6 h. Patients were selected on the basis of hyperglycaemia (>150 mg/dl) during complete parenteral nutrition with glucose (3.2+/-1.4 mg kg(-1) min(-1)), amino acids (n=8; 0.9+/-0.2 mg kg(-1) min(-1)), with or without lipid emulsions. Four hours (8 a.m. to 12 a.m. on parenteral nutrition plus NaCl as placebo) were compared to 4 h (12 a.m. to 4 p.m.) with additional GLP-1 administered intravenously. The dose of GLP-1 was 1.2 pmol kg(-1) min(-1). Blood was drawn for the determination of glucose, insulin, C-peptide, GLP-1, glucagon, and free fatty acids. RESULTS: Glycaemia was raised dose-dependently by glucose infusions in healthy volunteers (p<0.0001). GLP-1 ( approximately 100-150 pmol/l) stimulated insulin and reduced glucagon secretion and reduced glucose concentrations into the normoglycaemic fasting range (all p<0.05). In hyperglycaemic patients, glucose concentrations during the placebo period averaged 211+/-24 mg/dl. This level was reduced to 159+/-25 mg/dl with GLP-1 (p<0.0001), accompanied by a rise in insulin (p=0.0002) and C-peptide (p<0.0001), and by trend towards a reduction in glucagon (p=0.08) and free fatty acids (p=0.02). GLP-1 was well tolerated. CONCLUSIONS: Hyperglycaemia during parenteral nutrition can be controlled by exogenous GLP-1, e.g. the natural peptide (available today), whereas the chronic therapy of Type 2 diabetes requires GLP-1 derivatives with longer duration of action. PMID- 14759562 TI - Plasma adrenomedullin concentration is increased in patients with peripheral arterial occlusive disease associated with vascular inflammation. AB - Adrenomedullin (AM), a potent vasodepressor, is known to have anti atherosclerotic and anti-inflammatory effects. However, there is no information about its level in severe atherosclerotic diseases, such as peripheral arterial occlusive disease (PAOD). The present study investigated the plasma concentration of AM and several inflammatory parameters in 72 patients with and without PAOD. The plasma AM concentration in patients with PAOD was significantly higher than in those without PAOD. Its concentration had significant correlations with ankle brachial index and Fontaine's stage. The plasma AM level also correlated with high sensitive C-reactive protein and interleukin-6. As an additional study, plasma levels of two forms of AM drawn from the femoral artery and saphenous vein were measured in 27 other subjects. Both mature and intermediate forms of plasma AM in the femoral artery and saphenous vein were higher in patients with PAOD than in those without PAOD. A significant step-up of the mature form of AM from the femoral artery to the saphenous vein was observed. Our findings indicate that the plasma AM concentration was elevated in patients with PAOD in proportion to the severity of the disease and associated with vascular inflammation. An increased production of AM in PAOD may play a protective role against advanced atherosclerosis with an inflammatory signature. PMID- 14759563 TI - Influence of the lactokinin Ala-Leu-Pro-Met-His-Ile-Arg (ALPMHIR) on the release of endothelin-1 by endothelial cells. AB - Milk protein-derived peptides with angiotensin-converting enzyme (ACE) inhibitory activity can reduce blood pressure in hypertensive subjects. The lactokinin Ala Leu-Pro-Met-His-Ile-Arg (ALPMHIR) is an ACE-inhibitory peptide released by tryptic digestion from the milk protein beta-lactoglobulin. Its ACE-inhibitory activity is 100 times lower than that of captopril. The latter is known to inhibit the release of the vasoconstrictor endothelin-1 (ET-1) by endothelial cells. The effects of ALPMHIR on the endothelium are currently unknown. In this study, the influence of ALPMHIR on release of ET-1 by endothelial cells was investigated. The basal ET-1 release of the cells was reduced by 29% (p<0.01) in the presence of 1 mM ALPMHIR, compared to 42% (p<0.01) for 0.1 mM captopril. Addition of 10 U/ml thrombin to the incubation medium increased the release of ET 1 by 66% (p<0.01). Co-incubation of 10 U/ml thrombin with 1 microM captopril or with 0.1 mM ALPMHIR inhibited the stimulated ET-1 release by 45% (p<0.01) and by 32% (p<0.01), respectively. These data indicate that dietary peptides, such as ALPMHIR, can modulate ET-1 release by endothelial cells. These effects, among other mechanisms, may play a role in the anti-hypertensive effect of milk protein derived peptides. PMID- 14759564 TI - The cholecystokinin2-receptor mediates calcitonin secretion, gene expression, and proliferation in the human medullary thyroid carcinoma cell line, TT. AB - Gastrin-induced release of calcitonin from medullary thyroid carcinomas (MTC) is based on the expression of the cholecystokinin(2)-receptor (CCK(2)R) in these tumors. Recently, we have shown that the CCK(2)R is expressed not only in MTC but also in C-cells within the normal thyroid gland. The functions of the CCK(2)R in MTC and C-cells are largely unknown. We therefore explored the effects of gastrin induced CCK(2)R stimulation in the highly differentiated MTC cell line, TT. CCK(2)R expression in TT-cells is detectable by RT-PCR as well as immunocytochemistry. Stimulation of the CCK(2)R by gastrin induces immediate release of calcitonin from TT-cells. Moreover, quantitative (LightCycler) RT-PCR demonstrates that gastrin stimulates transcription of the calcitonin and chromogranin A genes in TT-cells. TT-cell proliferation, assessed by counting of viable cells and (3)H-thymidine uptake, is markedly increased by gastrin. This effect is inhibited by the CCK(2)R-specific antagonist L-365,260. Our findings suggest physiological functions for the CCK(2)R in calcitonin-secretion and gene expression as well as a pathophysiological role in MTC proliferation. CCK(2)R antagonists might have therapeutic potential in these tumors. PMID- 14759565 TI - Induction of heat shock protein 32 (Hsp32) in the rat cochlea following hyperthermia. AB - The genes for heat shock proteins (Hsps) can be upregulated in response to cellular trauma, resulting in enhanced cell survival and protection. Hsp32, also known as heme oxygenase 1, catalyzes the degradation of heme to produce carbon monoxide and bilirubin, which play a variety of cytoprotective functions at physiological concentrations, and iron, which is rapidly sequestered by the iron binding protein ferritin. In the present study we examined the expression and localization of Hsp32 in the rat cochlea after heat shock using semi-quantitative reverse transcription polymerase chain reaction (RT-PCR), Western blot, and immunocytochemistry. Low levels of constitutive Hsp32 expression were observed in the normal rat cochlea by RT-PCR and Western blot. Hsp32 mRNA (messenger RNA) was present at higher levels in a subfraction containing sensorineural epithelium and lateral wall than in a subfraction containing modiolus. Western blot revealed that Hsp32 protein levels increase in the rat cochlea following heat shock. Immunocytochemistry showed scattered staining of outer hair cells in the organ of Corti of normal untreated rats. Following heat shock Hsp32 is upregulated in outer hair cells and the cells of the stria vascularis. These results suggest a potential role for Hsp32 as a component of the oxidative stress response pathway in the rat cochlea. PMID- 14759566 TI - A periodic network of neurochemical modules in the inferior colliculus. AB - A new organization has been found in shell nuclei of rat inferior colliculus. Chemically specific modules with a periodic distribution fill about half of layer 2 of external cortex and dorsal cortex. Modules contain clusters of small glutamic acid decarboxylase-positive neurons and large boutons at higher density than in other inferior colliculus subdivisions. The modules are also present in tissue stained for parvalbumin, cytochrome oxidase, nicotinamide adenine dinucleotide phosphate-diaphorase, and acetylcholinesterase. Six to seven bilaterally symmetrical modules extend from the caudal extremity of the external cortex of the inferior colliculus to its rostral pole. Modules are from approximately 800 to 2200 microm long and have areas between 5000 and 40,000 microm2. Modules alternate with immunonegative regions. Similar modules are found in inbred and outbred strains of rat, and in both males and females. They are absent in mouse, squirrel, cat, bat, macaque monkey, and barn owl. Modules are immunonegative for glycine, calbindin, serotonin, and choline acetyltransferase. The auditory cortex and ipsi- and contralateral inferior colliculi project to the external cortex. Somatic sensory influences from the dorsal column nuclei and spinal trigeminal nucleus are the primary ascending sensory input to the external cortex; ascending auditory input to layer 2 is sparse. If the immunopositive modular neurons receive this input, the external cortex could participate in spatial orientation and somatic motor control through its intrinsic and extrinsic projections. PMID- 14759567 TI - Age-related hearing loss and the ahl locus in mice. AB - C57BL/6 (B6) mice experience hearing loss and cochlear degeneration beginning about mid-life, whereas CAST/Ei (CAST) mice retain normal hearing until old age. A locus contributing to the hearing loss of B6 mice, named age-related hearing loss (ahl), was mapped to Chromosome 10. A homozygous, congenic strain of mice (B6.CAST-+ahl ), generated by crossing B6 (ahl/ahl) and CAST (+ahl/+ahl) mice has the same genomic material as the B6 mice except in the region of the ahl locus, which is derived from CAST. In this study, we have determined the extent of the CAST-derived region of Chromosome 10 in the congenic strain and have examined mice of all three strains for hearing loss and cochlear morphology between 9 and 25 months of age. Results for B6 mice were similar to those described previously. CAST mice showed no detectable hearing loss even at 24 months of age; however, they had a small amount of ganglion cell degeneration. B6.CAST-+ahl mice were protected from early onset hearing loss and basal turn degeneration, but older animals did show some hearing loss and ganglion cell degeneration. We conclude that loci in addition to ahl contribute to the differences in hearing loss between B6 and CAST mice. These results illustrate the complex inheritance of age related hearing loss in mice and may have implications for the study of human presbycusis. PMID- 14759568 TI - Isoflurane/N2O anesthesia suppresses narrowband but not wideband inhibition in dorsal cochlear nucleus. AB - Anesthesia alters the response properties of neurons in the dorsal cochlear nucleus (DCN). Barbiturates decrease spontaneous activity and the prevalence of inhibitory responses, so that DCN principal cells show less inhibition by narrowband stimuli (e.g. tones at best frequency). Here we present the effects on cat DCN of anesthesia using isoflurane plus nitrous oxide (N2O). Because the cellular anesthetic mechanisms of isoflurane differ from those of pentobarbital, the effects of the two anesthetics in DCN might be different. The strength of two inhibitory circuits in the DCN, the narrowband and wideband inhibitor, were studied and compared with results in unanesthetized decerebrate animals. The primary effects of isoflurane/N2O anesthesia were to lower spontaneous activity and increase the thresholds of units. All the response types seen in the decerebrate preparation were also seen with isoflurane/N2O, but the prevalence of predominantly inhibitory responses to narrowband stimuli (type IV units) decreased (from approximately 31% to approximately 11%). However, responses to band-reject noise were similar to those seen in unanesthetized animals. Together, these results suggest that the effects of isoflurane/N2O are primarily on the narrowband inhibitory circuit, rather than the wideband inhibitor. PMID- 14759569 TI - Phenotypic variability of non-syndromic hearing loss in patients heterozygous for both c.35delG of GJB2 and the 342-kb deletion involving GJB6. AB - Mutations in GJB2, encoding the gap junction protein connexin 26, are the most common cause of inherited non-syndromic hearing loss (NSHL), with a broad spectrum of mutations leading to recessive as well as dominant forms. It has been shown that patients who are compound heterozygous for a 342-kb deletion (Delta(GJB6-D13S1830)) involving a large portion of the 5'-part of GJB6, encoding connexin 30, and a GJB2 mutation develop NSHL due to a trait with a digenic pattern of inheritance. We have used a mutation-specific polymerase chain reaction assay to screen NSHL patients for the presence of Delta(GJB6-D13S1830) and identified two families segregating both c.35delG in GJB2 and Delta(GJB6 D13S1830). Remarkably, the severity of hearing loss due to heterozygosity for c.35delG in GJB2 in conjunction with Delta(GJB6-D13S1830) is considerably different in members of the two families, ranging from congenital deafness in one to moderate/severe hearing loss with congenital onset in the other case. PMID- 14759570 TI - Noise induces A1 adenosine receptor expression in the chinchilla cochlea. AB - Adenosine plays a major cytoprotective role during ischemia and conditions of oxidative stress. Previous studies in our laboratory indicate that oxidative stress induces expression of the A1 adenosine receptor (A1AR) via activation of nuclear factor (NF)-kappaB. In this study, we tested whether noise exposure could induce oxidative stress and determine whether this induces expression of the A1AR in the chinchilla cochlea. Chinchillas were exposed to a 96 dB 4 kHz octave band of noise for 6 h of daily exposure, followed by an 18 h noise-free period. This noise paradigm resulted in threshold shifts of 10-60 dB over the frequency range (1-16 kHz) tested. Radioligand binding studies for the A1AR indicate a significant increase in receptor ( approximately 2-fold) expression soon after the first noise exposure period (usually within approximately 8 h of the initiation of noise), which gradually returned to basal levels by day 7. The rise in A1AR levels was followed by a significant increase in malondialdehyde levels by day 3, which also recovered by day 7. Assessment of the activity of NADPH oxidase in the cochlea indicates a significant increase in enzyme activity which was evident by approximately 8 h following initiation of noise exposure, and which persisted for at least up to day 3. Electrophoretic mobility shift assays indicate that the increase in A1AR was associated with a significant increase in NF-kappaB activity following noise exposure. We conclude that noise exposure induces A1AR expression, which might be mediated, in part, through generation of reactive oxygen species and activation of NF-kappaB. PMID- 14759571 TI - Cochlear delays measured with amplitude-modulated tone-burst-evoked OAEs. AB - Delay times in the mammalian cochlea, whether from measurement of basilar membrane (BM) vibration or otoacoustic emissions (OAEs) have, to date, been largely based on phase-gradient estimates from steady-state responses. Here we report cochlear delays measured directly in the time domain from OAEs evoked by amplitude-modulated tone-burst (AMTB) stimuli. Measurement using OAEs provides a non-invasive estimate of cochlear delay but is confounded by the complexity of generation of such OAEs. At low to moderate stimulus levels, and provided that the stimulus frequency range does not include a region of the cochlea where there is a large change in effective reflectance, AMTB stimuli evoke an OAE with an envelope shape that is similar to the stimulus and allow a direct calculation of cochlear group delay. Such delays are commensurate with BM estimates of delay, estimates of cochlear delay inferred from neural recordings, and previous OAE measures of delay in the guinea pig. However, a nonlinear distortion mechanism, variation in effective reflectance, and intermodulation distortion products generated by the nonlinear interaction in the cochlea of the carrier and sidebands of the AMTB stimulus, may all contribute to OAEs arising with envelope shapes that are not a scaled representation of the stimulus, confounding the estimation of cochlear group delay. PMID- 14759572 TI - A revised model of loudness perception applied to cochlear hearing loss. AB - We previously described a model for loudness perception for people with cochlear hearing loss. However, that model is incompatible with our most recent and most satisfactory model of loudness for normal hearing. Here, we describe a loudness model that is applicable to both normal and impaired hearing. In contrast to our earlier model for impaired hearing, the new model correctly predicts: (1) that a sound at absolute threshold has a small but finite loudness; (2) that, for levels very close to the absolute threshold, the rate of growth of loudness is similar for normal ears and ears with cochlear hearing loss; (3) the relation between monaural and binaural threshold and loudness; (4) recent measures of equal loudness contours. Like the earlier model, the new model can account for the loudness recruitment and reduced loudness summation that are typically associated with cochlear hearing loss. PMID- 14759573 TI - Morphometry of otoconia in the utricle and saccule of developing Japanese quail. AB - The development of otoconia in the utricular and saccular maculae from initial embryonic formation to adult stages was examined in Japanese quails. Both the morphology and size of the otoconia were quantified at different developmental stages. It was observed that the otoconia were initially formed on embryologic stage E5 in the saccule and E6 in the utricle. Otolith mass areas increased in a sigmoidal growth pattern, with saccular otolith areas being smaller than the utricular mass areas. Saccular otolith masses reached adult values at embryonic stage E12 and utricular areas reached adult values at post-hatch day 7. Mature individual otoconia were characterized by a barrel shape with two trihedral faceted ends. However, initial formation of otoconia at E5 (saccular) and E6 (utricular) maculae was characterized by a double fluted morphology that consisted of an hourglass shape with extended fins forming trihedral angles of 120 degrees. Double fluted otoconia rapidly filled, so that by embryonic day 8 mature otoconia dominated the maculae for the remainder of development through adulthood. Thus, a progression from double fluted to mature forms was noted. Mature utricular otoconia in adult quails averaged 11 microm in length and 5 microm in width, with length/width ratios of approximately 2.5:1, for all size ranges. Saccular otoconia were smaller, having about 70% the size of utricular otoconia in both length and width. During development, the average size and range of individual otoconia increased nearly linearly for both otolith organs. In the utricular macula, large otoconia were concentrated in the lateral regions of the epithelium. In contrast, otoconia of various sizes were distributed uniformly across the surface of the saccular macula. PMID- 14759574 TI - Further prospective findings with compound action potentials from Nucleus 24 cochlear implants. AB - The purpose of this study was to gain greater understanding of compound action potential (CAP) specific characteristics including: slope of the growth function, P1-N1 amplitude, threshold and latencies of P1 and N1 measured in cochlear implant users. Experienced adult subjects underwent behavioral threshold (T) measurement and electrically elicited stapedial reflex (eSR) recording, followed by CAP measurements on six selected electrodes. Based on the electrically elicited stapedial reflex threshold (eSRT), maximum stimulation level for each measured electrode was set. Relationships among the three thresholds of the above measures and maximum CAP P1-N1 amplitude and slope of the growth function were statistically evaluated for each measured electrode. Threshold of the CAP response showed relationships of similar strength with eSRT and T (r=0.69 and 0.61, respectively). For both slope of the growth function and CAP P1-N1 amplitude, a statistically significant relationship with cochlear place was found. Both specific characteristics of CAP measurement for the most apical electrodes were roughly double those for the most basal electrode (alpha=0.05). This may be partially explained by cochlear anatomy and is consistent with prior mammalian and human studies showing increasing density and survival of spiral ganglion cells in the regions corresponding to intracochlear electrode placement from basal to apical electrodes (90-360 degrees ). PMID- 14759575 TI - Effects of a calcium channel blocker on spontaneous neural noise and gross action potential waveforms in the guinea pig cochlea. AB - The effects of the L-type Ca2+ channel blocker nimodipine on the spectrum of the spontaneous neural noise (SNN) and the waveform of the gross sound-evoked compound action potential (CAP) were investigated by perilymphatic perfusion in the guinea pig cochlea. Both the SNN and the CAP were reversibly suppressed by nimodipine. The percentage reduction in SNN was dose-dependent in a manner very similar to the results obtained with the measures of CAP threshold changes. The reduction in the peak SNN caused by 10 microM nimodipine was the same as that caused by 500 microM kainic acid, which totally eliminated any neural responses. For 1 microM nimodipine there was an apparent dissociation between the SNN and CAP changes such that the SNN could be markedly suppressed with only very small changes in CAP thresholds. These results imply that spontaneous release of neurotransmitter from the inner hair cell is more sensitive to block of calcium channels than evoked release. There was no evidence for any marked shift caused by nimodipine, in the position of the main (900 Hz) spectral peak in the SNN. Comparison of the CAP waveform before and after nimodipine perfusion showed that the CAP waveforms were unchanged despite the change in sensitivity. These data do not support the notion of any significant postsynaptic site of action of nimodipine. The data hence provide further support for an exclusively presynaptic role for L-type Ca2+ channels in the regulation of both evoked and spontaneous neurotransmitter release from inner hair cells. PMID- 14759576 TI - Commentary: botulinum toxin in clinical medicine. AB - The recent introduction of botulinum toxin (BTX) into clinical medicine has revolutionized the practice of medicine and surgery forever. Who would have ever thought that the world's most deadly of all toxins would be used for therapeutic purposes? The discovery of the beneficial effects of BTX has transformed the lives of many unfortunate individuals who have suffered from a variety of unrelated disorders, which have been virtually impossible to treat. With the help of BTX, these and hopefully many other ailments will be easily managed with a few simple injections. PMID- 14759577 TI - History of the clinical use of botulinum toxin A and B. AB - The history of the cosmetic use of botulinum toxin (BTX) must begin with the clinical use of BTX in humans. This came out of Alan Scott's work, which was initially an attempt to find a nonsurgical treatment for some forms of strabismus. Alan Scott put together the idea of Ed Maumenee to use BTX to affect extraocular muscles, Art Jampolsky's electromyographic control delivery system, and Ed Schantz' work on the purification of BTX. PMID- 14759578 TI - Psychosocial aspects of beauty: how and why to look good. AB - It is difficult to pick up a paper or magazine today or turn on the television without being reminded that ours is a culture of youth and beauty. Bookstore shelves sag under the weight of enticing tomes devoted to the subject. We are bombarded with advertisements for any one of thousands of different products, both prescription and over-the-counter, that claim to be able to restore our youthful appearance, banish wrinkles, tone skin, and remove cellulite, among other promises that often sound, and probably are, too good to be true. Health food stores and sales catalogs are replete with myriad products, so that one wonders how the average person, in face of such bombardment, can either resist the temptation to buy or make an informed selection from the veritable feast of goodies put before them. Billions of dollars are expended each year, and false claims abound, but fortunately, there are many preparations and procedures that, in skilled hands, can really make a difference. PMID- 14759579 TI - Pharmacology and immunology of botulinum toxin type A. AB - The utility of botulinum neurotoxins as therapeutic and esthetic agents depends on their ability to inhibit neurotransmitter release from selected neurons, remain localized at the site of injection, and evade the body's immunologic defenses. The clinical correlates of these actions, respectively, are efficacy, safety, and a low rate of antibody formation. These properties have long formed the basis for the use of botulinum toxin type A (BTX-A) in the treatment of movement disorders such as focal dystonias, spasticity, and cerebral palsy and, more recently, in the treatment of glabellar lines--all of which are characterized by excessive muscle activity. PMID- 14759580 TI - Commercial preparations and handling of botulinum toxin type A and type B. AB - The recognition of botulinum toxin as a therapeutic tool has revolutionized the approach to facial esthetics. In the 1970s, Dr. Alan B. Scott pioneered the use of botulinum toxin type A (BTX-A) for strabismus. This discovery led to further studies examining other clinical applications of BTX-A and, more recently, botulinum toxin type B (BTX-B). Despite its widespread use and the more than two decades of experience with BTX-A, controversy remains over the preparation and handling of the toxin. PMID- 14759581 TI - Patient management and treatment technique. AB - In contrast to the direct effect of augmentation materials, the smoothing effect on folds and wrinkles after a botulinum toxin treatment (BTX) is a reactive, secondary effect after achieving reduction of muscle activity first. Besides the relaxing effect on mimic muscles and overlying skin, the treatment leads to psychological effects as well. The interruption of long-established, reflexlike muscle hyperactivity or even pathologic muscle activity (tics) changes not only the patient's personal perception of his environment and the impression he leaves on his environment but also his entire aspect. PMID- 14759582 TI - Type A botulinum toxin in the upper aspect of the face. AB - The use of botulinum toxin type A (BTX-A) for facial rejuvenation was first systematically developed by Carruthers and Carruthers on the upper aspect of the face. In the early 1990s, the first studies of the cosmetic use of BTX-A were published. BTX-A was only approved in 2002, however, by the U.S. Food and Drug Administration when extensive cosmetic studies were evaluated. The treatment of facial wrinkles with BTX-A has truly changed the concepts held by physicians regarding facial rejuvenation, mainly on the upper aspect of the face. It is one of the most common cosmetic procedures currently performed by physicians. PMID- 14759583 TI - Periocular botulinum toxin. AB - Botulinum toxin type A (BOTOX, Dysport) has revolutionized treatment of wrinkles around the eyes. Since the first publications of its cosmetic benefit by Drs. Jean and Alastair Carruthers, hundreds of articles have been published about its cosmetic use. BOTOX holds U.S. Food and Drug Administration approval in the United States for treatment of glabellar lines. BOTOX is in widespread use worldwide and is currently the most popular cosmetic treatment in the United States. PMID- 14759585 TI - Botulinum toxin for rejuvenation of the neck. AB - The use of botulinum toxin for cosmetically restoring an aging neck is a simple procedure that has great patient satisfaction. It is also a safe and effective therapeutic modality for any patient desiring rejuvenation of his or her neck, but who at the same time does not desire any recuperation time or is unwilling to undertake the potential risks associated with plastic surgery. Currently, both botulinum toxin type A (BTX-A) and B are being used to treat the aging neck with great success. However, our clinical experience has been more vast with BTX-A, and the accomplishment of several clinical trials with it has yielded more precise results. BTX-A can be used to correct early signs of aging as well as more advanced forms. It helps correct jowl formation and platysmal banding as well as horizontal rhytides in the neck. PMID- 14759584 TI - Botulinum toxin in the lower one third of the face. AB - The use of botulinum toxin (BTX) in the lower one third of the face has rapidly become one of the most popular ways to rejuvenate the face, due to the effectiveness of BTX in this area and the lack of major side effects. It is recommended that physicians planning on treating patients in this area of the face first have extensive experience treating the more traditional areas, such as the forehead, glabella, or crow's feet, because the inferior one third of the face has many co-dependent and interdigitating (interrelated) muscles that are very sensitive and readily affected by minimal amounts of BTX. We prefer to reconstitute a 100-U vial of Botox(R) with 1-2.5 mL of saline solution because some of the muscles in this area of the face are very sensitive to diffusion of the toxin and will be weakened beyond the area originally planned for treatment. PMID- 14759586 TI - How to avoid complications when treating hyperdynamic folds and wrinkles. AB - Before discussing the complications that might occur in the treatment of hyperdynamic folds and wrinkles, it is important to analyze and define the correct effects and unwanted effects, or even adverse reactions, after administration of botulinum toxin (BTX) for cosmetic indications. The desired therapeutic effect is a result of a relaxation or even paralysis of the target muscle. Both intended and adverse effects are self-limited. Because this is a cosmetic treatment, not only severe adverse reactions but also patient dissatisfaction are the subjects of the following discussion. PMID- 14759587 TI - Colin Andrew Ramsay, MD, FRCP (June 11, 1936-April 24, 2003). PMID- 14759588 TI - From hormones to immunity: the physiology of immunology. AB - Discoveries in the physiology of immunology have increased at an increasing rate during the past two decades. It is now recognized that the immune system is just another physiological system that regulates, and is regulated by, other physiological systems such as the brain. These advances make it clear that recent findings in genomic biology must be interpreted in the context of the environment in which animals and humans live. Lack of a strong genetic basis for significant human mental health disorders, such as major depression, points to the critical importance of interactions. Several examples of environmental x genetic x disease interactions are presented. Regulation of cells of the hematopoietic lineage by two genes that control over 80% of postnatal growth, growth hormone and IGF-I, are then highlighted. The reciprocal relationship of how proinflammatory cytokines from the immune system regulate the growth hormone/IGF-I axis is also summarized. Particular emphasis is placed upon TNFalpha-induced IGF-I resistance in neurons, muscle cells and epithelial cells. This cytokine regulation of hormone action may ultimately be more important for human and animal health than direct effects of growth hormone and IGF-I on hematopoietic cells. Wasting of AIDS patients is given as an important clinical example of how TNFalpha from an activated immune system reduces IGF-I sensitivity in multiple physiologic systems, including muscle, nervous and hematopoietic tissues. PMID- 14759589 TI - Neuroimmune networks and communication pathways: the importance of location. PMID- 14759590 TI - Reversible inactivation of the dorsal vagal complex blocks lipopolysaccharide induced social withdrawal and c-Fos expression in central autonomic nuclei. AB - Peripheral administration of lipopolysaccharide (LPS), a potent activator of the immune system, induces symptoms of behavioral depression, such as social withdrawal, concommitant with increases in c-Fos expression in central autonomic network nuclei. Previous studies implicated vagal visceral sensory nerves in transduction of immune-related signals relevant to for the induction of social withdrawal, a symptom of behavioral depression. Vagal sensory nerves terminate in the dorsal vagal complex (DVC) of the brainstem, a region that functions to integrate visceral signals and may also play a role in modulating arousal and affect. The objective of the current study was to determine whether the DVC contributes to immunosensory pathways driving symptoms of social withdrawal associated with LPS-induced behavioral depression, using a reversible lesion technique to temporarily inactivate the DVC. To assess the effects of DVC inactivation on LPS-induced social withdrawal and the subsequent changes in brain activation, we used behavioral assessment of social withdrawal, and analyzed c Fos expression, a marker of neuronal activation, in the central nucleus of the amygdala (CEA), bed nucleus of the stria terminalis (BST), hypothalamic paraventricular nucleus (PVN), and ventromendial preoptic area (VMPO). Two hours following intraperitoneal LPS injection, there was a significant increase in c Fos immunoreactivity in forebrain regions in animals treated with LPS. DVC inactivation completely blocked LPS-induced social withdrawal and dramatically reduced LPS-induced Fos expression in all four forebrain regions assessed. Collectively, these findings support the idea that the DVC acts as an immune behavior interface between the peripheral stimuli and brain areas involved in modulating social behavior. PMID- 14759591 TI - Immune consequences of the spontaneous pro-inflammatory status in depressed elderly patients. AB - INTRODUCTION: The aim of the study was to describe the interrelationship between senescence, depression, and immunity. METHODS: We assessed 10 elderly patients with depression and 10 age- and sex-matched controls: before, at one and at six month intervals after the anti-influenza vaccination. Levels of TNFalpha, IL6, ACTH, and cortisol, titres of anti-hemagglutinins and anti-neuraminidases, lymphocytes secreting IFNgamma, IL2, IL4, and IL10, cytotoxicity of NK and CD3+ CD8+ IFNgamma+ cells, anti-CMV antibodies, and CD28- CD57+ lymphocytes known to be associated with the CMV carrier status were evaluated. RESULTS: Higher levels of anti-CMV, higher percentage of the CD28- CD57+ cells, and elevated levels of TNFalpha, IL6, and cortisol concomitant with decreased levels of ACTH and insufficient production of IL10 (which increased the IFNgamma+ /IL10+ ratio) were found in the patients suffering from depression, in comparison to healthy controls. The subjects with depression revealed a low NK cytotoxicity, while a level of CD3+ CD8+ IFNgamma+ cells was comparable between the groups. Although the levels of anti-hemagglutinins and anti-neuraminidases were low in the depressed patients, they reached the protective titres. The majority of these differences disappeared when CMV titres were entered into the analyses as a covariate. DISCUSSION: The results suggest that the elderly depressed patients were characterised by increased exposure to CMV in the past, which could have resulted in a pro-inflammatory profile demonstrated as elevated levels of TNFalpha, IL6 and deficiency of suppressive IL10+ cells. These changes negatively affect humoral and innate response in the depressed patients. PMID- 14759592 TI - Alpha-tocopherol attenuates lipopolysaccharide-induced sickness behavior in mice. AB - Antioxidants protect cells from oxidative damage and reduce lipopolysaccharide (LPS)-induced expression of inflammatory cytokines. Because inflammatory cytokines induce sickness behavior, we hypothesized that antioxidants, namely alpha-tocopherol (alpha-T) and selenium would inhibit sickness behavior caused by LPS. In an initial study, mice were injected intraperitoneal (i.p.) with vehicle, 2, or 20mg alpha-T for 3 consecutive days and then challenged with vehicle, 1, 10, or 100 microg of LPS. Sickness behavior was quantified by measuring social exploratory behavior. Vehicle pretreated mice injected with LPS showed a marked reduction in social behavior at 4h (p < .01). However, sickness behavior induced by the lowest dose of LPS was partially or completely blocked by 2 or 20mg alpha T, respectively. alpha-T did not prevent sickness from higher doses of LPS. In a second study, mice were fed AIN93-M modified diets containing 10, 75, and 500 mg alpha-T/kg and 0.05, 0.15, and 2mg selenium/kg for 8 weeks and then challenged with saline or LPS (1 microg). The highest concentration of dietary alpha-T and selenium tended (p = .1) to reduce LPS-induced sickness behavior. Mice fed diets low in antioxidants had reduced plasma alpha-T levels and glutathione peroxidase activity (p = .08 and p < .01, respectively) and elevated liver thiobarbituric acid reactive substances (p < .001) 24h post LPS. Collectively, these data indicate that alpha-T improved the oxidative status after exposure to LPS, which may explain its ability to ameliorate symptoms of sickness. PMID- 14759593 TI - The modulator role of the hypothalamic paraventricular nucleus on immune responsiveness. AB - Role of the paraventricular nucleus of the hypothalamus (PVH) upon immune modulation was studied by either mechanically destroying the PVH (PVHL) or by isolating the PVH (PVHI) with a knife-cut. PVHL or PVHI manipulations induced significant leukopenia characterized by a decrease in the number of neutrophils and lymphocytes two weeks post surgery. The numbers of circulating monocytes and eosinophils were not affected by PVH interventions. In addition, PVHL and PVHI were also associated with a reduction, relative to controls, in the phagocytosis by neutrophils and an increase in blastic transformation of T lymphocytes induced by phytohemagglutinin-M (PHA-M). Antibody titers rose against sheep red blood cells (SRBC) after either PVHL or PVHI were reduced. The magnitude of the SRBC antibody reduction after PVH manipulations was similar to that observed in rats that received a peripheral chemical sympathectomy two hrs prior immunization. Comparison of thyroid hormones blood levels two weeks after PVHL or PVHI revealed significant reductions in comparison with sham-operated group (SO), whereas blood corticosterone was not significantly altered. In summary, we provide evidence that lesion or isolation of the PVH selectively reduces circulating white blood cells and the primary immune response, while it enhances the cell-mediated immune function. Taken together our data showed that PVH modulates immune functions by altering both the peripheral sympathetic tone and thyroid hormone secretion. PMID- 14759594 TI - The effects of restraint stress on the neuropathogenesis of Theiler's virus infection II: NK cell function and cytokine levels in acute disease. AB - Psychological stress is thought to play an important role in multiple sclerosis. We have been investigating the role of restraint stress in Theiler's virus infection in mice as a model for multiple sclerosis. We have previously determined that restraint stressed CBA mice had higher levels of mortality following infection with Theiler's virus. We proposed that this was due to high levels of stress-induced corticosterone, which resulted in decreased numbers of circulating lymphocytes, decreased inflammatory cell infiltrates into the brain and consequently decreased viral clearance from the central nervous system (CNS). The effect of restraint stress on the innate immune response to Theiler's virus is further investigated in the current study. Restraint stressed mice developed clinical signs of encephalitis, thymic atrophy, and adrenal hypertrophy. Decreased numbers of circulating lymphocytes and increased numbers of neutrophils were observed in the stressed mice. Stressed mice also had lower numbers of spleen cells which correlated with the decreased numbers of lymphocytes in circulation. Restraint stress caused elevations in serum tumor necrosis alpha (TNF-alpha). Virus-induced natural killer cell (NK) cytotoxic activity was significantly reduced in restrained mice at one day post infection which may account for the reduced viral clearance from the CNS. These data suggest that stress-induced immunosuppression of cytolytic NK cell activity may account in part for the reduced ability to clear virus from the CNS and increased mortality observed in this model. PMID- 14759595 TI - Kainic acid-induced excitotoxic hippocampal neurodegeneration in C57BL/6 mice: B cell and T cell subsets may contribute differently to the pathogenesis. AB - The roles of T cells and B cells in kainic acid (KA)-induced hippocampal lesions were studied in C57BL/6 mice lacking specific T cell populations (CD4, CD8, and CD4/CD8 cells) and B cells [Igh-6(-/-)]. At 48 mg/kg of KA administrated intranasally, KA-induced convulsions were seen in all groups. However, CD4/CD8(-/ ) mice exhibited the mildest seizures; the responses of CD8(-/-), Igh-6(-/-) and wild-type mice were intermediate, whereas CD4(-/-) mice displayed much more severe clinical signs and 100% early mortality, indicating that a deficiency of CD4 T cells obviously increased susceptibility to KA-induced brain damage. Histopathological analysis of the mice that survived 7 days after KA administration revealed that CD4/CD8(-/-) mice had the fewest pathologic changes but Igh-6(-/-) mice showed more severe lesions in area CA3 of the hippocampus than CD8(-/-) and wild-type mice. Reactive astrogliosis were prominent in all KA treated mice. Locomotor activity as assessed by open-field test increased after KA administration in Igh-6(-/-) and wild-type mice only. These results denote the influence of the adaptive immune response on KA-induced hippocampal neurodegeneration and suggest that B cell and T cell subsets may contribute differently to the pathogenesis. PMID- 14759596 TI - Conditioned taste aversion with lipopolysaccharide and peptidoglycan does not activate cytokine gene expression in the spleen and hypothalamus of mice. AB - Several reports show that behavioural and physiological components of the acute phase reaction can be conditioned. However, the mechanisms responsible for these effects remain obscure. The underlying assumption that the changes observed in conditioned animals are dependent on a conditioned production of cytokines has never been demonstrated. In the present study, the possibility of conditioning the production of cytokines or molecules implicated in their signalling pathways was tested by submitting mice to conditioned taste aversion with a new saccharin taste paired with intraperitoneal (i.p.) injections of lipopolysaccharide (LPS, 0.83 microg/g) or peptidoglycan (PGN, 20 microg/g). After two conditioning sessions, conditioned mice developed a clear aversion to saccharine that was not associated with activation of genes of the cytokine network either at the periphery, or in the hypothalamus, as demonstrated by a macroarray approach and confirmed by real time RT-PCR. In contrast, there was an activation of the genes coding for nuclear factor kappa B (NFkappaB) and mitogen activated protein kinase (MAPK) signalling pathways in the spleen and to a lesser extent in the hypothalamus. This modulation of the NFkappaB and MAPK signalling pathways is interpreted in terms of a possible conditioned sensitisation of the immune system. PMID- 14759598 TI - An assessment of proposed mechanisms for sensing hydrogen peroxide in mammalian systems. AB - Despite much recent interest in the biochemistry of reactive oxygen species, the mechanisms by which hydrogen peroxide (H2O2) functions in mammalian cells remain poorly defined. Proposed mechanisms for sensing H2O2 in mammalian cells include inactivation of protein tyrosine phosphatases and dual specificity phosphatases as well as inactivation of peroxiredoxins. In this critical review, proteins proposed to serve as sensors for H2O2 in mammals will be compared to peroxidases, catalases, and the bacterial H2O2 sensor OxyR for their ability to react with H2O2, in the context of our current knowledge concerning the concentrations of H2O2 present in cells. PMID- 14759599 TI - Azide binding to yeast cytochrome c peroxidase and horse metmyoglobin: comparative thermodynamic investigation using isothermal titration calorimetry. AB - Yeast cytochrome c peroxidase (CcP) and horse metmyoglobin (Mb) bind HN3 with similar affinities at 25 degrees C. The pH-independent equilibrium association constants for formation of the CcP.HN3 and Mb.HN3 complexes are (1.05 +/- 0.06)x10(5) and (1.6 +/- 0.8)x10(5) M(-1), respectively. However, the thermodynamic parameters for formation of the two complexes are quite different. The DeltaH0 values for formation of CcP.HN3 and Mb.HN3 are -16.4 +/- 0.7 and -9.0 +/- 0.5 kcal/mol, respectively, and the Delta S0 values are -32 +/- 2 and -16 +/- 2 cal/deg mol, respectively. The proton associated with HN3 is retained in both protein complexes at low pH but dissociates with apparent pKA values of 5.5 +/- 0.2 and > or =8.2 for the Mb.HN3 and CcP.HN3 complexes, respectively. CcP and Mb differ significantly in their reactivity toward the azide anion, N3-. CcP binds N3- very weakly, if at all, and only an upper-limit of 18 +/-5 M(-1) for the pH independent equilibrium association constant for the CcP.N3- complex can be determined. Mb binds N3- with an association constant of (1.8 +/- 0.1)x10(4) M( 1). The ratio of the equilibrium association constants for HN3 and N3- binding provides a discrimination factor between the neutral and charged forms of the ligand. The discrimination factor is greater than 5800 for CcP but only nine for Mb. Protonation of the distal histidines in the two proteins influences binding of HN3. Protonation of His-64 in Mb enhances HN3 binding due to a gating mechanism while protonation of His-52 in CcP decreases the affinity for HN3 due to loss of base-assisted association of the ligand to the heme iron. PMID- 14759600 TI - Overexpression of DRG2 suppresses the growth of Jurkat T cells but does not induce apoptosis. AB - Developmentally regulated GTP-binding protein (DRG) is a new subfamily within the superfamily of GTP-binding proteins. Its expression is regulated during embryonic development. To investigate the effect of the expression of DRG2 on cell growth, we constructed a human Jurkat-T-cell line that overexpresses DRG2. Overexpression of DRG2 suppressed the growth and the aggregation of Jurkat cells but did not induce apoptotic cell death. We used cDNA microarray analysis to examine the global changes in gene expression induced by an overexpression of DRG2. DNA array analyses identified genes that may suppress cell growth at a number of levels in multiple signaling cascades in Jurkat cells and also several prosurvival genes that may protect cells from apoptosis. PMID- 14759601 TI - Discrimination of depolarized from polarized mitochondria by confocal fluorescence resonance energy transfer. AB - Mitochondrial depolarization promotes apoptotic and necrotic cell death and possibly other cellular events. Polarized mitochondria take up cationic tetramethylrhodamine methylester (TMRM), which is released after depolarization. Thus, TMRM does not label depolarized mitochondria. To identify both polarized and depolarized mitochondria in living cells, cultured rat hepatocytes, and sinusoidal endothelial cells were co-loaded with green-fluorescing MitoTracker Green FM (MTG) and red-fluorescing TMRM for imaging by laser scanning confocal microscopy. Like TMRM, MTG is a cationic fluorophore that accumulates electrophoretically into polarized mitochondria. Unlike TMRM, MTG binds covalently to intramitochondrial protein thiols and remains bound after depolarization. In cells labeled only with MTG, excitation with blue (488 nm) light yielded green but almost no red fluorescence. After subsequent loading with TMRM, green MTG fluorescence became quenched. Instead, blue excitation yielded red fluorescence. Mitochondrial de-energization restored green fluorescence and abolished red fluorescence. Conversely, when MTG was added to TMRM-labeled cells, red fluorescence excited by blue light was enhanced, an effect again reversed by de-energization. These observations of reversible quenching of donor fluorescence and augmentation of acceptor fluorescence signify fluorescence resonance energy transfer (FRET). In undisturbed hepatocytes, spontaneous depolarization of a subfraction of mitochondria was an ongoing phenomenon. In conclusion, confocal FRET discriminates individual depolarized mitochondria against a background of hundreds of polarized mitochondria. PMID- 14759602 TI - Nuclear shuttling of the peptidase nardilysin. AB - The metalloendopeptidase nardilysin contains a putative N-terminal nuclear localization signal. The functionality of this sequence was tested with nardilysin-GFP fusion constructs. Expression in NIH3T3 cells showed approximately 90-95% of nardilysin-GFP as cytoplasmic. However, 3-6% of transfected cells showed both cytosolic and nuclear staining, while 2-4% showed predominantly nuclear staining. A nuclear localization signal mutant and an N-terminally truncated nardilysin-GFP with the nuclear localization signal deleted were completely cytoplasmic. Although endogenous nardilysin was barely detectable in the nucleus, after treatment with leptomycin B, nuclear nardilysin rose to approximately 15% and to over 25% after addition of spermine. The ability of a methionine 49 to act as the sole initiator methionine, as previously proposed, was tested by inserting a c-myc epitope between leucine28 and glycine29. Expression in HEK293 cells showed the presence of the c-myc tag, demonstrating that the enzyme can be translated from the first methionine and contains the nuclear localization signal. PMID- 14759603 TI - Effect of substrate mechanics on chondrocyte adhesion to modified alginate surfaces. AB - This study characterized the attachment of chondrocytes to RGD-functionalized alginate by examining the effect of substrate stiffness on cell attachment and morphology. Bovine chondrocytes were added to wells coated with 2% alginate or RGD-alginate. The alginate was crosslinked with divalent cations ranging from 1.25 to 62.5 mmol/g alginate. Attachment to RGD-alginate was 10-20 times higher than attachment to unmodified alginate and was significantly inhibited by antibodies to integrin subunits alpha3l and beta1, cytochalasin-D, and soluble RGD peptide. The equilibrium level and rate of attachment increased with crosslink density and substrate stiffness. Substrate stiffness also regulated chondrocyte morphology, which changed from a rounded shape with nebulous actin on weaker substrates to a predominantly flat morphology with actin stress fibers on stiffer substrates. The dependence of attachment on integrins and substrate stiffness suggests that chondrocyte integrins may play a role in sensing the mechanical properties of the matrices to which they are attached. PMID- 14759604 TI - 20S proteasome mediated degradation of DHFR: implications in neurodegenerative disorders. AB - The 20S proteasome is responsible for the degradation of protein substrates implicated in the onset and progression of neurodegenerative disorders, such as alpha-synuclein and tau protein. Here we show that the 20S proteasome isolated from bovine brain directly hydrolyzes, in vitro, the dihydrofolate reductase (DHFR), demonstrated to be involved in the pathogenesis of neurodegenerative diseases. Furthermore, the DHFR susceptibility to proteolysis is enhanced by oxidative conditions induced by peroxynitrite, mimicking the oxidative environment typical of these disorders. The results obtained suggest that the folate metabolism may be impaired by an increased degradation of DHFR, mediated by the 20S proteasome. PMID- 14759605 TI - Roles of Munc18-3 in amylase release from rat parotid acinar cells. AB - Several "soluble N-ethylmaleimide-sensitive fusion protein attachment protein receptor" (SNARE) proteins have been identified in rat parotid acinar cells, including VAMP-2, syntaxin 4, and SNAP-23. Furthermore, an association between Munc18c (Munc18-3) and syntaxin 4 has been reported. However, the role of Munc18 3 in secretory granule exocytosis on parotid acinar cells remains unclear. In the present study, we investigated the role of Munc18-3 in rat parotid acinar cells. Munc18-3 was localized on the apical plasma membrane where exocytosis occurs and interacted with syntaxin 4. Anti-Munc18-3 antibody dose-dependently decreased isoproterenol (IPR)-induced amylase release from SLO-permeabilized parotid acinar cells. Furthermore, stimulation of the acinar cells with IPR induced translocation of Munc18-3 from the plasma membrane to the cytosol. Munc-18-3 was not phosphorylated by a catalytic subunit of protein kinase (PK) A but phosphorylated by PKC. Treatment of the plasma membrane with PKC but not PKA induced displacement of Munc18-3 from the membrane. The results indicate that Munc18-3 regulates exocytosis in the acinar cells for IPR-induced amylase release and that phosphorylation of Munc18-3 by PKA is not involved in the mechanism. PMID- 14759606 TI - Platelet storage under in vitro condition is associated with calcium-dependent apoptosis-like lesions and novel reorganization in platelet cytoskeleton. AB - Platelets are cleared from circulation after a life span of 8-10 days. The molecular mechanisms underlying platelet senescence remain poorly characterized. Here we report that, progressive functional impairment in the platelets incubated in vitro in a plasma-free isotonic medium for up to 24 h at 37 degrees C is associated with release of cytochrome c from platelet mitochondria and cleavage of procaspase-9, but without evidence of caspase-3 activation. Concomitantly, there was proteolysis of survival proteins like focal adhesion kinase, Src, gelsolin, and specific cytoskeleton-associated peptides, in a manner regulated by extracellular calcium and calpain activity. Cytoskeleton played a critical role as evidenced from the association of these proteins and their degradation products, as well as procaspase-3 and the actin regulatory small GTPase, CDC42Hs, with the cytoskeleton of the stored platelets. The cytoskeletal enrichment with specific proteins was not associated with increase in the content of F-actin and was cytochalasin-resistant, thus signifying a novel mechanism of interaction of the translocating proteins with the pre-existing cytoskeleton. There was progressive exposure of phosphatidylserine on the outer leaflet of platelet membrane and specific electron microscopic changes suggestive of apoptotic lesions. Based on these observations we discuss the caspase-independent but calpain-mediated signaling events in the stored platelets resembling the features of apoptosis in the nucleated cells. PMID- 14759607 TI - All three isoforms of the voltage-dependent anion channel (VDAC1, VDAC2, and VDAC3) are present in mitochondria from bovine, rabbit, and rat brain. AB - All three isoforms of the voltage-dependent anion channel (VDAC) were detected by immunoblot analysis of mitochondria isolated from rat, rabbit, and bovine brain. All three isoforms were associated with mitochondria after fractionation of rat brain extracts on sucrose density gradients. No VDAC isoforms were detected in non-mitochondrial fractions. Relative levels of the mRNAs coding the VDAC isoforms in rat, rabbit, and bovine brain were determined by RT-PCR. In all three species, the mRNA for VDAC2 was predominant. Relative to the mRNA for VDAC3, mRNAs for both VDAC1 and VDAC2 were more highly expressed in bovine brain than in rat brain. These results are consistent with the possibility that differences in relative expression of VDAC isoforms may be a factor in determining the species dependent ratio of Type A:Type B hexokinase binding sites on brain mitochondria. PMID- 14759608 TI - Catalase transgenic mice: characterization and sensitivity to oxidative stress. AB - The role of catalase in the antioxidant defense system was studied using transgenic mice [Tg(CAT)] harboring a human genomic clone containing the entire human CAT gene. Catalase activity was 2-fold higher in the tissues of hemizygous [Tg(CAT)(+/o)] mice and 3- to 4-fold higher in the tissues of homozygous [Tg(CAT)(+/+)] mice compared to wild type mice. The human CAT transgene was expressed in a tissue-specific pattern that was similar to the endogenous catalase gene. The levels of other major antioxidant enzymes were not altered in the tissues of the transgenic mice. Hepatocytes and fibroblasts from the Tg(CAT)(+/+) mice were more resistant to hydrogen peroxide-induced cell death but were more sensitive to paraquat and TNFalpha toxicity. Fibroblasts from the Tg(CAT)(+/+) mice showed reduced growth rate in culture without treatment and reduced colony-forming capability after gamma-irradiation compared to fibroblasts from wild type mice. In addition, the Tg(CAT)(+/+) animals were more sensitive to gamma-irradiation. PMID- 14759609 TI - Posttranslational processing of polysaccharide lyase: maturation route for gellan lyase in Bacillus sp. GL1. AB - Cells of Bacillus sp. GL1 extracellularly secrete a gellan lyase with a molecular mass of 130 kDa responsible for the depolymerization of a heteropolysaccharide (gellan), although the gene is capable of encoding a huge protein with a molecular mass of 263 kDa. A maturation route for gellan lyase in the bacterium was determined using anti-gellan lyase antibodies. The fluid of the bacterial exponentially growing cultures on gellan contained two proteins with molecular masses of 260 and 130 kDa, both of which reacted with the antibodies. The 260 kDa protein was purified from the cultured fluid and characterized. The protein exhibited gellan lyase activity and showed similar enzyme properties, such as optimal pH and temperature, thermal stability, and substrate specificity, to those of the 130 kDa gellan lyase. The N-terminal amino acid sequences of the 260 and 130 kDa enzymes were found to be identical. Determination of the C-terminal amino acid of the 130 kDa enzyme indicated that the 260 kDa enzyme is cleaved between the 1205Gly and 1206Leu residues to yield the mature form (130 kDa) of the gellan lyase. Therefore, the mature enzyme consists of 1170 amino acids (36Ala-1205Gly) with a molecular weight of 125,345, which is in good agreement with that calculated from SDS-PAGE analysis. Judging from these results, gellan lyase is first synthesized as a preproform (263 kDa) and then secreted as a precursor (260 kDa) into the medium through cleavage of the signal peptide. Finally, the precursor is post-translationally processed into the N-terminal half domain of 130 kDa as the mature form, the function of C-terminal half domain being unclear. PMID- 14759610 TI - Single chain antibodies that recognize the N-glycosylation site. AB - We aimed to identify antibodies that can recognize the Asn-Xaa-Ser/Thr(NXS/T) N glycosylation site that guides oligosaccharyltransferase (OT) activity. We used synthetic Asn-Cys-Ser/Thr(NCS/T) tripeptides conjugated to bovine serum albumin to isolate single chain antibody fragments of a variable region (scFv) from the Griffin 1 phage antibody library. Although Ser and Thr have different side chains, the scFv proteins thus isolated bound to both NCS and NCT with Kd values of the order of 10(-6) M and accepted the substitution of the Cys residue with various amino acids, including Ala, Gly, and Val. However, these proteins recognized neither Asn-Pro-Ser/Thr nor non-NXS/T tripeptides. The scFv proteins recognized NCS/T and N-glycosylation site of mutant yeast protein disulfide isomerase when they were in their native but not denatured state. These results indicate that antibody recognition of the NXS/T motif is conformation dependent and suggest that NXS/T spontaneously adopts a specific conformation that is necessary for antibody recognition. These features are likely to correlate with the known binding specificity of OT. PMID- 14759611 TI - Making research matter. PMID- 14759612 TI - Progestogen-only pills and high blood pressure: is there an association? A literature review. AB - The progestogen-only pill (POP) is a contraceptive option for women who have high blood pressure either induced by use of combined oral pills or due to other causes; as long as it is well controlled and monitored. Combined oral contraception (COC) and Depo-Provera have been implicated in increased cardiovascular risk following use. High blood pressure has been theorized to be the critical path that leads to this increased risk. POP is the recommended method for women who are at risk of coronary heart disease due to presence of risk factors like hypertension. In order to offer POP as a safe, alternative contraception to women who develop hypertension on COCs or those who are at increased cardiovascular risk, it is important to take into account evidence of no association of high blood pressure with POP use. A search of published medical literature (PUBMED and Cochrane database) was undertaken with this objective. A total of four articles were selected for final review after application of inclusion and exclusion criteria. Three of these were prospective control trials and one a cross-sectional survey. There was no randomized study to answer this question. The results of these studies consistently reported no significant association of high blood pressure with use of POPs for up to 2-3 years of follow up. PMID- 14759613 TI - The association between depot medroxyprogesterone acetate contraception and bone mineral density in adolescent women. AB - Although adolescent women are actively acquiring bone, there has been little study of the possible effects of depot medroxyprogesterone acetate (DMPA) injectable contraception use on bone density in adolescents. We conducted a cross sectional evaluation of the association between DMPA use and bone mineral density in adolescent women, ages 14-18 years. Of 174 study participants, 81 were DMPA users (range, 1-13 injections, median = 3) and 93 were not. Mean bone density at all anatomic sites (hip, spine and whole body) was lower for DMPA users than nonusers, but differences were not statistically significant (e.g., hip, 0.940 vs. 0.970 g/cm2, p = 0.10; spine, 0.970 vs. 0.992 g/cm2, p = 0.19). Duration of DMPA use showed a trend toward lower spine bone density (p-value for trend = 0.06). This study did not find a strong association between DMPA use and bone density. Further prospective evaluation of bone density changes with DMPA use and after DMPA discontinuation are needed in this age group. PMID- 14759614 TI - Double-blind, randomized study comparing the effects of two monophasic oral contraceptives containing ethinylestradiol (20 microg or 30 microg) and levonorgestrel (100 microg or 150 microg) on lipoprotein metabolism. AB - The effects of two monophasic oral contraceptives containing ethinylestradiol 20 microg in combination with levonorgestrel 100 microg (EE20/LNG100) or 30 microg and 150 microg (EE30/LNG150), respectively, on lipoprotein metabolism was investigated in a double-blind, randomized study of 12 treatment cycles in healthy female volunteers. Total triglycerides (+32% to +46%, p < 0.05 in comparison to baseline) increased significantly. Triglycerides were highest after six cycles of treatment, decreasing thereafter. Total cholesterol (+1% to +7%), apolipoprotein (apo) B (+21% to +29%) and low-density lipoprotein (LDL) cholesterol (+7% to +17%) increased slightly. High-density lipoprotein (HDL) cholesterol decreased slightly (-11% and -5%), HDL triglycerides increased (+16% and +26%). Apo AI did not change during the study, suggesting that the molar concentration of HDL particles did not change. Apo E (-23% to -14%) decreased, and there was a transitory decrease of lipoprotein (a). Essentially, there was no difference regarding the changes in lipoprotein metabolism between the two treatment groups. The effects of the two combinations of ethinylestradiol and levonorgestrel on triglyceride-rich lipoproteins appear less pronounced than those produced by preparations containing third-generation progestins. It is not likely that the changes in lipoprotein metabolism brought about by the two preparations will alter the risk of future cardiovascular disease in a clinically relevant fashion. PMID- 14759615 TI - Menstrual pattern and lipid profiles during use of medroxyprogesterone acetate and estradiol cypionate and NET-EN (200 mg) as contraceptive injections. AB - The objectives of this study were to compare effects of medroxyprogesterone acetate 25 mg + estradiol cypionate 5 mg (Cyclofem) and norethisterone enanthate (NET-EN) upon the menstrual pattern and determine changes in lipoprotein parameters after 12 months of use. One-hundred females were included and 87 (45 with Cyclofem and 42 with NET-EN) women completing 12 months were evaluated. Menstrual changes were the leading complaint among users. At the end of 12 months, 20/45 (44.4%) and 18/41 (43.9%) Cyclofem and NET-EN users, respectively, had normal menstrual pattern. Irregular and infrequent bleeding were the two most important changes that occurred. The discontinuation rate at 12 months due to menstrual disturbances did not show any significant differences between the two preparations, but showed lower incidence compared to other studies. Total cholesterol, high-density, low-density and very low-density lipoprotein cholesterol and triglyceride levels decreased at 12 months in both groups and these changes were statistically significant. PMID- 14759616 TI - Cervical changes associated with progestagen-only contraceptives: a team approach. AB - OBJECTIVES: To report clinical, cytologic and colposcopic findings among women using progestagen-only contraceptives for more than 3 years as compared to nonhormonal contraceptives; and to assess the role of nursing in increasing the women's knowledge about Pap test and risk factors for cervical cancer in a developing country set-up. DESIGN: A prospective cross-sectional comparative study. SETTING: Family-planning clinic of a tertiary care university hospital. SUBJECTS: A total of 325 current users of contraceptive methods for more than 3 years. INTERVENTIONS: Patients were divided into two groups. Group A included 200 patients using progestagen-only contraceptives. While group B comprised 125 patients wearing intrauterine devices. In both groups, the patient was asked about the risk factors and her knowledge about cancer cervix. Clinical and cytologic evaluations of the cervix were done. For each patient, two cervical smears were taken using Ayre's spatula and the endocervical brush. MAIN OUTCOME MEASURES: Clinical, cytologic and histopathologic cervical abnormalities after prolonged use, and the effect of health education or counseling on the patient's knowledge about preinvasive or invasive cervical cancer. RESULTS: Initial naked eye assessment of the cervix revealed statistically significant difference between both groups (p < 0.000). However, cytologic examinations revealed positive cases of low- and high-grade squamous intraepithelial lesions (SIL) in 38 (19%) and 22 (17.6%) in the study and control groups, respectively, without any statistically significant difference. There were no statistically significant differences in both groups regarding the frequency of positive cases whether using Ayre's spatula or the endocervical brush. Women's knowledge was compared in the first and last interviews by the nursing staff involved in health education or counseling. It increased in the counseling group; however, the difference was statistically insignificant. CONCLUSIONS: Prolonged use of progestagen-only contraceptives is not associated with increased risk of abnormal cytologic findings. There was a marked lack of women's knowledge as regards Pap test and cervical cancer with some statistically insignificant improvement after interview and counseling. Extended training of the nursing staff on the procedure of Pap smear would help establish screening programs in the developing countries. PMID- 14759617 TI - The contraceptive vaginal ring, NuvaRing, and antimycotic co-medication. AB - We investigated the effect of antimycotic co-medication on the systemic exposure to etonogestrel (ENG) and ethinylestradiol (EE) released from the contraceptive vaginal ring, NuvaRing. Different formulations of miconazole nitrate and single as well as multiple dosing were investigated during two separate randomized, open label, crossover studies. The first study recruited 12 women to compare the effects of co-use of NuvaRing and a single dose of antimycotic to NuvaRing alone. The second study recruited 14 women to compare the effects of multiple doses of an antimycotic vaginal suppository to an antimycotic vaginal cream equivalent. Co administration of all three antimycotic formulations resulted in a slight increase in systemic exposure to ENG and EE over time, with suppositories having a more pronounced effect than a cream formulation in the multiple-dosing study. The increases in serum levels observed with the different antimycotic formulations are not expected to compromise NuvaRing's contraceptive efficacy or tolerability. PMID- 14759618 TI - Effect of hormonal emergency contraception on bleeding patterns. AB - Spotting following the use of emergency contraception is not unusual, nor is anxiety in women waiting to see if the treatment has worked. It is not known whether such spotting should bring worry or relief. We, therefore, wished to see if there was any correlation between bleeding pattern and treatment outcome. Using data from a large multicenter efficacy trial, we examined bleeding patterns post-emergency contraception. The earlier in the cycle the pills were taken, the more likely the next bleed was to be early and the less likely it was to be on time. There was no observable difference in spotting rates between women who got pregnant and those who did not. The occurrence of spotting did not influence whether the next period was lighter or heavier. PMID- 14759619 TI - Two-year performance of a Nestorone-releasing contraceptive implant: a three center study of 300 women. AB - A 2-year trial of a single Nestorone (NES) rod implant was conducted at three Latin American centers, each enrolling 100 women. We studied the safety, effectiveness and acceptability of this progestin-releasing contraceptive implant. Three pregnancies occurred, the last at 18 months of use. Because no pregnancies were expected in the first 18 months, the trial was halted. At that time, 224 women had completed at least 18 months of use, and 99 women had used the implant for more than 24 months. Few participants used adjunctive contraception between the time the study was halted and the time they had their implant removed. No additional pregnancies occurred before the removal of the last implant. The 2-year cumulative pregnancy rate was 1.7 per 100 with a Pearl index of 0.6 per 100 for the 2-year period. The 1-year and 2-year continuation rates were 80.5 and 66.7 per 100, respectively. Menstrual and medical disturbances were the principal reasons for discontinuation, followed by planned pregnancy. Headache and weight gain frequently led to discontinuation. The NES implant had little important effect on most clinical chemistry and lipid parameters. Over the study course, the mean change in hemoglobin was <1%. Slight modification of the design of this single 2-year implant, restoring features previously examined in clinical trials, is likely to improve its effectiveness. A single NES implant appears to provide acceptable contraception for women. PMID- 14759620 TI - Prospective randomized, controlled study of postoperative pain after titanium silicone rubber clip or Silastic ring tubal occlusion. AB - OBJECTIVE: To determine if laparoscopic sterilization using the Filshie clip causes less postoperative pain as compared to the Falope ring during the first 24 h after surgery. STUDY DESIGN: Forty subjects were enrolled. Five milliliters of 0.5% bupivicaine was injected at each of the two incision sites and 5 mL was dripped onto each fallopian tube. Subjects then received one Filshie clip and Falope ring on opposite fallopian tubes; the side of occlusion was randomized. Subjects were questioned about pain prior to the first administration of analgesics postoperatively, 1 and 2 h after surgery, at discharge, and 24 h after surgery. RESULTS: Forty subjects completed the study; 32 without major deviations from protocol. There was no significant difference in pain between sides with the Filshie clip or Falope ring at any times evaluated. CONCLUSION: There was no significant reduction in the postoperative pain associated with the Filshie clip as compared to the Falope ring within 24 h after surgery. PMID- 14759621 TI - Utero-ovarian morphology and blood flow after tubal ligation by the Pomeroy technique. AB - OBJECTIVES: The objectives of this study were to evaluate ovarian sonographic morphology and pelvic blood flow in patients who had undergone bilateral tubal ligation by the Pomeroy technique. DESIGN: Twenty women who had undergone bilateral tubal ligation during cesarean section by the Pomeroy techniques were compared to 20 matched controls who had undergone cesarean section alone. Ovarian morphology as well as ovarian and uterine pulsatility index (PI), resistance index and peak systolic velocity were evaluated using transvaginal sonography and Doppler velocimetry. RESULTS: Patients post tubal ligation by the Pomeroy technique were found to have a significantly higher rate of polycystic appearance of the ovaries (60% vs. 5%; p < 0.005). Six patients (30%) had post tubal ligation symptoms, as compared to none of the controls. Five of the six symptomatic tubal ligation patients (83%) had ovarian polycystic appearance, which was not significantly different than asymptomatic patients. Ovarian artery PI was significantly higher in symptomatic patients (2.7 +/- 1.9) compared to asymptomatic patients (1.5 +/- 0.9) (p < 0.05). CONCLUSIONS: We therefore conclude that patients post bilateral tubal ligation by the Pomeroy technique may acquire an ovarian polycystic appearance and increased ovarian PI, which may be associated with a tendency to develop post tubal ligation symptoms. PMID- 14759622 TI - Differences in efficacy, differences in providers: results from a hazard analysis of medical abortion. AB - Sample sizes of even the largest medical abortion trials are generally not adequate to provide an understanding of how well the regimen works for subgroups of women, particularly when controlling for factors known to influence efficacy, such as gestational age. By pooling data from four previously published studies of medical abortion and using hazard analyses, we can undertake such an investigation. We find that women with lower gestational ages, women younger than 23 years of age, women with more than 12 years of education and women with no previous induced abortion experience were more likely to experience a successful medical abortion. After taking into account demographic factors, we find that significant differences in efficacy persist across study sites, indicating that differences in providers' tendency to intervene by performing vacuum aspiration vary across medical abortion providers. PMID- 14759623 TI - Sublingual compared with oral misoprostol for cervical dilatation prior to vacuum aspiration: a randomized comparison. AB - The aim of the study was to compare the effect of oral and sublingual administration of misoprostol for cervical priming prior to vacuum aspiration. Thirty-two first-time-pregnant women with 8-12 weeks amenorrhea were randomly assigned to receive 400 microg misoprostol either orally or sublingually 3 h prior to surgery. The degree of baseline dilatation and the cumulative force needed for dilatation of the cervical canal did not differ between the two treatment groups. However, the number of patients in whom a strong force was needed was significantly higher following oral than following sublingual treatment. It was shown that sublingual administration is more effective than oral administration of misoprostol for cervical priming and is associated with less blood loss but a higher frequency of side effects. PMID- 14759624 TI - Which cannulae fit the Ipas manual vacuum aspiration syringe? AB - BACKGROUND: The Ipas double-valve manual vacuum aspiration (MVA) syringe is an alternative to electrical vacuum aspiration. METHODS: This study determines which US cannulae (flexible, rigid-straight and rigid-curved) work with the Ipas MVA syringe. Cannulae from Ipas, MedGyn, Berkeley and Milex, in sizes 6-12 mm, were randomized and affixed to the MVA syringe. A pressure gauge was attached to the cannula with rubber tubing. Pressure readings were recorded initially and over 30 s. RESULTS: Milex cannulae were not compatible. For the remaining brands, initial vacuum pressures ranged from 55 to 65 mmHg. Flexible cannulae from Ipas, Berkeley and MedGyn maintained initial pressures without leaks, as did the 6-12-mm straight- and curved-rigid cannulae by Berkeley. Eight of the 13 tested MedGyn rigid cannulae lost >10% pressure over 30 s. CONCLUSION: Several US manufacturers produce cannulae that fit on the Ipas MVA syringe without a leak, including Ipas flexible cannula; Berkeley flexible, rigid-straight and rigid-curve cannulae and MedGyn flexible cannula, but not their rigid cannulae. PMID- 14759625 TI - Evaluating multiple sclerosis. PMID- 14759626 TI - Correlations between multiple sclerosis functional composite, expanded disability status scale and health-related quality of life during and after treatment of relapses in patients with multiple sclerosis. AB - The measurement of the clinical manifestations of multiple sclerosis (MS) is difficult. In the present study, we examined the changes in measurement of functions during and after pulse methylprednisolon (MP) treatment of MS exacerbations using the MSFC and EDSS. Correlation between multiple sclerosis quality of life (MSQoL)-54, EDSS and MSFC were studied. Thirty-six clinically definite MS patients were included in this study. Because of MSFC's repeating feature, we administered the tests to a control group to exclude practise effects. All patients received 1000-mg intravenous MP for 5 days, followed by tappering dose of 100-mg oral prednisolone. All three scales were assessed on day 0. EDSS and two components of MSFC (nine HPT and T25WT) were administered on the other days of pulse MP treatment. PASAT was not applied before the day 5 to exclude the practise effect. MSQoL-54 was assessed again on day 30. Mean EDSS values significantly decreased after the day 2. MSFC score improved from 0.03 +/- 1.71 on day 0 to 0.79 +/- 1.51 on day 5. Improvement continued on day 30. The mean physical health composite score increased from 66.50 +/- 9.3 on day 0 to 74.34 +/- 8.9 on day 30. Mental health composite had also a significant improvement on day 30. Correlation between the baseline overall MSFC and the EDSS was moderately strong. T25WT correlated most strongly with EDSS. Significant positive correlation was found between MSFC and both components of MSQoL-54. It is more prominent for the MSFC and physical health composite correlation. The same correlation was found for the EDSS and MSQoL-54 composites. Changes in EDSS and MSFC scores and MSQoL-54 were found significantly correlated for the overall score on day 30 compared with day 0. In conclusion, MSFC seems to be more sensitive in detecting changes in function than the EDSS. Hence, EDSS is still useful for daily routine practise. When these results combined with the significant correlation between MSFC and MSQoL-54 measures, which indicated the MSFC reflects the severity of MS as perceived by patients, MSFC seems to be the most useful scale for clinical trials. PMID- 14759627 TI - Functional imaging during covert auditory attention in multiple sclerosis. AB - Recent literature suggests that the brain in multiple sclerosis (MS) undergoes reorganization that subserves the performance of visual and motor tasks. We identified sites of cerebral activity in 16 MS patients while performing a covert attention (CA) task, presented in the auditory modality. Positron emission tomography (PET) revealed activation of rostral/dorsal anterior cingulate cortex (ACC) in normal subjects studied previously. Activity in this region was not significant in MS patients, but there was a large region of activity in superior temporal cortex. Decreased activation of frontal attentional networks and greater activity in sensory/perceptual cortical areas (auditory association cortex) suggests a reduction of transmission along white matter tracts connecting these regions. This study demonstrates cingulate hypoactivity and cerebral reorganization during auditory attention in MS. PMID- 14759628 TI - Oxidative stress in Alzheimer's and vascular dementias: masking of the antioxidant profiles by a concomitant Type II diabetes mellitus condition. AB - Oxidative stress is associated with Alzheimer's (DAT) and vascular (VD) dementias, as well as Type II diabetes mellitus (DIAB) and affected by hypoglycemic therapy. The population (n = 122; males = 60; mean age = 72.57 +/- 7.06) consisted of controls (CTR), DAT and VD patients, with (DAT + DIAB, VD + DIAB) and without concomitant DIAB, resulting in six groups where the antioxidant profile was determined: copper-zinc superoxide dismutase (SOD), thiobarbituric acid reactive substances (TBARS), and total antioxidant capacity (TRAP). The results were analyzed using a two-way ANOVA design and Bonferroni statistic. The ANOVAs yielded significant differences between groups for all components of the profile: SOD, p = 0.00000006; TBARS, p = 0.0000012; TRAP, p = 0.0000003. The significance level for comparisons between groups was set at alpha = 0.05. The comparisons DIAB vs. CTR, DAT+DIAB vs. DAT, and DIAB demented vs. DIAB non demented resulted significant for all variables. VD + DIAB vs. VD resulted significant for all variables except TRAP. The antioxidant profiles of DIAB and CTR are different. The differences cannot be directly related with what is observed in dementias. The differences in profiles of demented and non-demented are somewhat hidden when demented patients are affected by a concomitant DIAB condition and/or hypoglycemic treatment, thus conditioning the diagnostic value for dementias of the profiles. PMID- 14759629 TI - Investigation of a neuronal nitric oxide synthase gene (NOS1) polymorphism in a multiple sclerosis population. AB - Multiple Sclerosis (MS) is a chronic neurological disease characterized by demyelination associated with infiltrating white blood cells in the central nervous system (CNS). Nitric oxide synthases (NOS) are a family of enzymes that control the production of nitric oxide. It is possible that neuronal NOS could be involved in MS pathophysiology and hence the nNOS gene is a potential candidate for involvement in disease susceptibility. The aim of this study was to determine whether allelic variation at the nNOS gene locus is associated with MS in an Australian cohort. DNA samples obtained from a Caucasian Australian population affected with MS and an unaffected control population, matched for gender, age and ethnicity, were genotyped for a microsatellite polymorphism in the promoter region of the nNOS gene. Allele frequencies were compared using chi-squared based statistical analyses with significance tested by Monte Carlo simulation. Allelic analysis of MS cases and controls produced a chi-squared value of 5.63 with simulated P = 0.96 (OR(max) = 1.41, 95% CI: 0.926-2.15). Similarly, a Mann Whitney U analysis gave a non-significant P-value of 0.377 for allele distribution. No differences in allele frequencies were observed for gender or clinical course subtype (P > 0.05). Statistical analysis indicated that there is no association of this nNOS variant and MS and hence the gene does not appear to play a genetically significant role in disease susceptibility. PMID- 14759630 TI - Neuropsychological functions in variant Alzheimer's disease with spastic paraparesis. AB - Few data exist on the effects of specific Alzheimer's disease (AD)-related mutations on cognitive function. We present neuropsychological test results in eight members of a large kindred with variant Alzheimer's disease (VarAD) due to a deletion of the presenilin 1 (PS-1) gene, encompassing exon 9. The disease was neuropathologically characterized by the presence of large, unusual, "cotton wool" plaques (CWP). Four surviving patients were prospectively tested, and retrospective neuropsychological data were collected from additional four deceased patients. The neuropsychological evaluation was based on tests of verbal and visual memory, abstract thinking, and visuoconstructive and spatial functions. In addition, psychiatric symptoms were evaluated. In four patients, brain glucose metabolism was examined by positron emission tomography (PET). PET showed temporoparietal hypometabolism typical of AD. In addition, variable patterns of hypometabolism (hemispherical asymmetry and occipital accentuation) were related to individual deficits of cognitive performance. However, all these early-onset patients (age range 43-63 years) with a deletion mutation of PS-1 gene showed prominent memory impairment and deficits in visuoconstructive and intellectual functions. PMID- 14759631 TI - Elevated levels of matrix metalloproteinases-9 and -1 and of tissue inhibitors of MMPs, TIMP-1 and TIMP-2 in postmortem brain tissue of progressive supranuclear palsy. AB - We determined the levels and tissue localization of matrix metalloproteinases (MMPs) as well as their endogenous tissue inhibitors (TIMPs) in postmortem brain tissue from 13 patients with progressive supranuclear palsy (PSP) and 8 age matched controls. MMP-9 expression was significantly increased in both the frontal cortex (p = 0.002) and substantia nigra (p = 0.003) of PSP cases as compared to controls whereas MMP-1 levels were increased in the substantia nigra (p = 0.01) but unchanged in the frontal cortex (p = 0.41). Levels of the endogenous tissue inhibitors of MMPs, TIMP-1 and TIMP-2 were significantly elevated in the substantia nigra (TIMP-1: p = 0.004, TIMP-2: p = 0.01). Levels of TIMPs were unchanged in PSP frontal cortex as compared to control cases. Together, these data show alterations of MMPs and TIMPs in the substantia nigra as well as in the frontal cortex of PSP, consistent with the possibility that alterations in MMPs/TIMPs may contribute to disease pathogenesis. PMID- 14759632 TI - Angiotensin-converting enzyme and alpha-2-macroglobulin gene polymorphisms are not associated with Alzheimer's disease in Colombian patients. AB - Polymorphisms in alpha-2-macroglobulin (A2M) and angiotensin-converting enzyme (ACE) genes have been considered as risk factors for Alzheimer's disease (AD) independently of the risk conferred by the apolipoprotein E sigma4 allele (APOEsigma4) in diverse populations. In the present study, we have analysed the distribution of genotypes and alleles of the insertion/deletion (I/D) polymorphisms of ACE and A2M in 83 AD patients and 69 normal controls in Colombia. Our results showed that there is no association between the I/D polymorphisms of ACE and A2M with AD (P = 0.788 and P = 0.538, respectively). Using logistic regression and multiple correlation analysis (MCA), we confirmed that the main risk factor associated and consistently grouped with AD patients in this population is APOE4, but this association was not observed with alleles and genotypes of ACE and A2M. PMID- 14759633 TI - Hyperornithinemia, hyperammonemia, and homocitrullinuria syndrome with evidence of mitochondrial dysfunction due to a novel SLC25A15 (ORNT1) gene mutation in a Palestinian family. AB - Hyperornithinemia, hyperammonemia, and homocitrullinuria (HHH) syndrome is caused by mutations in the SLC25A15 (ORNT1) gene encoding the mitochondrial ornithine transporter, but the mechanism of pathogenesis of the encephalopathy, spastic paraparesis and hepatopathy remains undetermined. HHH syndrome was diagnosed in a 2-year-old Palestinian boy with developmental delay and seizures, and subsequently in his 13-year-old brother with developmental delay. Direct sequencing of the PCR products of SLC25A15 exon amplifications revealed that both brothers were homozygous for a novel 446G deletion in exon 3 as well as for a 760A>T (I254L) polymorphism in exon 5, which is downstream of a premature termination codon produced by the frameshift resulting from the 446G deletion. The index patient had elevated liver enzymes as well as hyperalaninemia, lactic acidemia with an elevated lactate to pyruvate ratio, and increased urinary excretion of lactate, glutarate and Krebs cycle intermediates. These findings are indicative of mitochondrial dysfunction and are in accordance with ultrastructural studies showing increased numbers of large and bizarre mitochondria in liver, muscle, leukocytes and fibroblasts of some HHH patients. Neurologic and hepatic manifestations are characteristic of some primary mitochondrial disorders. Secondary mitochondrial dysfunction may contribute to the pathogenesis of these same features in HHH syndrome. PMID- 14759635 TI - Lower urinary tract dysfunction in Machado-Joseph disease: a study of 11 clinical urodynamic observations. AB - OBJECTIVES: Machado-Joseph disease (MJD), or hereditary spinocerebellar ataxia type 3, is the most common dominantly inherited ataxia. However, lower urinary tract (LUT) dysfunction in MJD has not been fully delineated. We investigated LUT dysfunction in MJD by clinical-urodynamic observations. METHODS: In 24 genetically diagnosed MJD, we recruited all 11 patients with LUT symptoms (six men, five women; age, 18-61 [mean 48] years; disease duration, 2-24 [mean 9] years; voiding difficulty, 7, urinary incontinence, 4). Urodynamic studies consisted of uroflowmetry, measurement of post-void residuals and electromyography (EMG)-cystometry. Neurophysiology tests consisted of motor unit potential (MUP) analysis of the sphincter and extremity muscles, tibial nerve somatosensory evoked potentials (SEP) and nerve conduction studies (NCS) of the extremities. RESULTS: Urodynamic abnormalities were seen in all 11 patients studied. Maximum or average flow rate was decreased in five. Post-void residual was noted in three but residual urine volume > 100 ml was noted in only one patient. Maximum urethral closure pressure was low in one and high in one of five patients studied. EMG-cystometry during filling showed detrusor overactivity in five, impaired bladder sensation in four, low compliance detrusor in one, uninhibited sphincter relaxation in one and incompetent urethra in one. Voiding phase abnormalities included detrusor areflexia in three and detrusor-sphincter dyssynergia in two. Bethanechol supersensitivity of the bladder was noted in one of three patients studied. Bulbocavernosus reflex was absent in two of five patients studied. MUP analysis showed neurogenic changes in six of nine sphincter muscles and in all six extremity muscles studied. Five patients had prolonged or absent cortical response in SEP and four had sensory axonal neuropathy in NCS, which were relevant to the impaired bladder sensation. CONCLUSION: In the present study, a half of MJD patients had LUT symptoms and they showed various urodynamic abnormalities. Detrusor overactivity, impaired bladder sensation, and neurogenic sphincter EMG were common findings, and large post-void residuals were rare. These findings are relevant to central and peripheral nervous system pathology of MJD. PMID- 14759634 TI - The use of complementary and alternative medicines by patients with peripheral neuropathy. AB - Complementary and alternative medicine (CAM) therapies have become increasingly popular and are used regularly by patients with chronic neurological disorders. The prevalence and characteristics of CAM use by patients with peripheral neuropathy is unknown. We performed a prospective, questionnaire-based study to determine the prevalence and patterns of use of CAM therapies in 180 consecutive outpatients with peripheral neuropathy. The use of CAM was reported by 77 patients (43%) with neuropathy. The most frequent were megavitamins (35%), magnets (30%), acupuncture (30%), herbal remedies (22%), and chiropractic manipulation (21%); 37 (48%) tried more than one form of alternative treatment. Seventeen respondents (27%) thought their neuropathy symptoms improved with these approaches. Those who used CAM were slightly younger (mean age 62 vs. 65 years, p = 0.05) and more often college educated (39% vs. 24%, p = 0.03) compared to CAM nonusers. They also more often reported burning neuropathic pain (62% vs. 44%, p = 0.01). Patients with diabetic neuropathy used CAM more frequently than others (p = 0.03). The most common reason for using CAM was inadequate pain control (32%). Almost half of patients did not consult a physician before starting CAM. We conclude that there is a high prevalence of CAM use in our patients with neuropathy, and one-quarter reported that their symptoms improved. CAM users were better educated than nonusers, but most did not discuss CAM treatments with their physician. Neuropathic pain was substantially more common in CAM users, and lack of pain control was the most common reason for CAM use. PMID- 14759636 TI - Treatment of progressive forms of multiple sclerosis by cyclophosphamide: a cohort study of 490 patients. AB - There are no generally effective disease-modifying drugs for progressive forms of multiple sclerosis (MS). Some MS centres use cyclophosphamide (CYC) in secondary progressive (SP) forms of MS, especially after interferon beta-1b (INFbeta-1b) treatment failure. Moreover, there are currently no approved drugs for primary progressive (PP) MS. Using the collected data of patients with progressive MS, we studied clinical patterns that predicted a good response to CYC treatment. Secondly, we compared the therapeutic response of SPMS and PPMS patients to the treatment. Data from 490 MS patients were collected. All patients presented an SP (n = 362) or PP (n = 128) form of the disease and 476 had been treated for at least one year with a monthly pulse of CYC associated with methylprednisolone (MP). CYC treatment was justified because of at least a 1-point worsening on the Expanded Disability Status Scale (EDSS) during the previous year. The EDSS score was assessed at baseline and after 6 months (M6) and 12 months (M12) of treatment. After 12 months of CYC treatment, 78.6% of SPMS and 73.5% of PPMS patients had stabilised or had an improved EDSS score. Response to CYC was not significantly different in the two progressive forms of MS. Twenty-two patients presented noticeable drug side effects, one of whom withdrew from the treatment due to intolerance. Patients with an improved EDSS at M12 had a shorter mean progressive time course (5.1 years) than patients who stabilised or worsened (7.1 years) (p = 0.02). We also observed that poor responders at M6 were also poor responders at M12 (p < 0.001). This large cohort study showed that CYC treatment was well tolerated and suggested that a better response occurred in cases with a short progressive time course. We did not find any difference in treatment response between the two progressive forms of MS. To date, no treatment is approved for PPMS and we therefore propose a trial to test the use of CYC treatment early in the course of the disease in PPMS patients with disability progression. PMID- 14759637 TI - Identification of two novel mutations in the Cu/Zn superoxide dismutase gene with familial amyotrophic lateral sclerosis: mass spectrometric and genomic analyses. AB - Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disorder affecting motor neurons. The majority of patients are sporadic cases, while 5-10% of the patients have a family history of ALS (fALS). Mutations in the gene that encodes cytoplasmic Cu/Zn superoxide dismutase (SOD1) have been identified in about 25% of fALS cases. Although the precise pathogenesis of ALS is still unknown, experimental studies including animal models suggest that fALS is caused by the toxic gain-of-function of the SOD1 mutant. We have analyzed not only SOD1 gene mutation by genomic sequencing, but also SOD1 mutant protein by liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS). We analyzed 33 fALS patients and found 10 mutations in SOD1 gene, in which two were novel: Asp101His substitution in exon 4 and Gly141Glu substitution in exon 5. Here, we present their mass spectrometric protein analyses and clinical features. PMID- 14759638 TI - Clinical analysis of longstanding subacute myelo-optico-neuropathy: sequelae of clioquinol at 32 years after its ban. AB - One thousand and thirty-one longstanding patients with subacute myelo-optico neuropathy (SMON; 275 males, 756 females; mean age +/- S.D., 72.9 +/- 9.6 years; age at onset 37.6 +/- 9.8 years; duration of illness 35.3 +/- 4.0 years) were examined in 2002, 32 years after banning of clioquinol. At onset, 66.7% of patients were unable to walk, and 4.7% complete blindness. At present time, about 41% of patients were still difficult to walk independently, including 15.8% of completely loss of locomotion. One point six percent of patients were in complete blindness and 5.8% had severe visual impairment. The majority (95.6 - 97.7%) of patients exhibited sensory disturbances including superficial and vibratory sensations and dysesthesia. Dysautonomia was observed as leg hypothermia in 79.8%, urinary incontinence in 60.7%, and bowel disturbance in 95.3%. As complication, high incidence was revealed with cataract (56.2%), hypertension (40.2%), vertebral disease (35.5%), and limb articular disease (31.5%). These results indicate the serious sequelae of clioquinol intoxication, SMON. PMID- 14759639 TI - Therapeutic effects of normal cells on ABCD1 deficient cells in vitro and hematopoietic cell transplantation in the X-ALD mouse model. AB - Bone marrow transplantation (BMT) is accepted as an efficient therapy for X linked adrenoleukodystrophy (ALD). To clarify the mechanisms of this treatment, we examined the effects of hematopoietic cell transplantation (HCT) in an ATP binding cassette, subfamily D, member 1 (ABCD1) knock out mice and co-culture of ALD patient fibroblasts with normal cells. We treated ABCD1 knock out mice with HCT using lacZ-transgenic mice as donors, which enabled us to detect donor derived cells. We also examined the effects of co-culturing a normal microglia cell line (N9) with ALD fibroblasts. beta-Galactosidase (beta-GAL) activity was higher in spleen, lung and kidney than in liver, brain and spinal cord of the recipient ABCD1 knock out mice. HCT reduced the accumulation of very long chain fatty acid (VLCFA) in those tissues. The reduction of the VLCFA ratio was significant in spleen and lung; tissues with higher beta-GAL activity. ABCD1 was detectable in spleen from HCT mice. Co-culture of ALD fibroblasts with normal fibroblast cells reduced VLCFA accumulation in ALD cells. This effect was not observed when the cells were co-cultured while separated by a filter membrane. Our data suggest that supplying normal cells for ABCD1 knockout mouse by HCT corrects metabolic abnormalities in ALD tissues through a cell-mediated process. The correction requires direct cell-to-cell contact for recovering normal cell function. PMID- 14759640 TI - No evidence for paternal inheritance of mtDNA in patients with sporadic mtDNA mutations. AB - With the publication of a patient with severe exercise intolerance, in whom the mutated mtDNA in muscle was shown to be paternally inherited, the strict maternal inheritance of mtDNA was challenged. Paternal mtDNA inheritance may have gone unrecognized in cases of mitochondrial disease with no clear maternal pattern of inheritance because mitochondrial haplotypes are rarely investigated in diagnostic analyses. To find further evidence for a paternal inheritance of mtDNA, we reinvestigated 12 patients with mitochondrial myopathy, in whom the pathogenic mutation was known to be sporadic. We compared the mtDNA haplotypes from the patient's muscle with that of the mtDNA haplotypes in blood from either the mother or the patient. No evidence of paternal inheritance of mtDNA was found in this small study. Although these findings indicate that the paternal inheritance of mtDNA is rare, they do not rule out that the phenomenon may occur at a rate that could still affect genetic counselling and anthropological research. PMID- 14759641 TI - Comparison of the effects of acetyl L-carnitine and amantadine for the treatment of fatigue in multiple sclerosis: results of a pilot, randomised, double-blind, crossover trial. AB - Treatment with acetyl L-carnitine (ALCAR) has been shown to improve fatigue in patients with chronic fatigue syndrome, but there have been no trials on the effect of ALCAR for treating fatigue in multiple sclerosis (MS). To compare the efficacy of ALCAR with that of amantadine, one of the drugs most widely used to treat MS-related fatigue, 36 MS patients presenting fatigue were enrolled in a randomised, double-blind, crossover study. Patients were treated for 3 months with either amantadine (100 mg twice daily) or ALCAR (1 g twice daily). After a 3 month washout period, they crossed over to the alternative treatment for 3 months. Patients were rated at baseline and every 3 months according to the Fatigue Severity Scale (FSS), the primary endpoint of the study. Secondary outcome variables were: Fatigue Impact Scale (FIS), Beck Depression Inventory (BDI) and Social Experience Checklist (SEC). Six patients withdrew from the study because of adverse reactions (five on amantadine and one on ALCAR). Statistical analysis showed significant effects of ALCAR compared with amantadine for the Fatigue Severity Scale (p = 0.039). There were no significant effects for any of the secondary outcome variables. The results of this study show that ALCAR is better tolerated and more effective than amantadine for the treatment of MS related fatigue. PMID- 14759643 TI - Application of SVM to predict membrane protein types. AB - As a continuous effort to develop automated methods for predicting membrane protein types that was initiated by Chou and Elrod (PROTEINS: Structure, Function, and Genetics, 1999, 34, 137-153), the support vector machine (SVM) is introduced. Results obtained through re-substitution, jackknife, and independent data set tests, respectively, have indicated that the SVM approach is quite a promising one, suggesting that the covariant discriminant algorithm (Chou and Elrod, Protein Eng. 12 (1999) 107) and SVM, if effectively complemented with each other, will become a powerful tool for predicting membrane protein types and the other protein attributes as well. PMID- 14759642 TI - Implications for hyperhomocysteinemia: not homocysteine but its oxidized forms strongly inhibit neuronal network activity. AB - Severe hyperhomocysteinemia (50-200 microM) often presents itself with acute neuronal dysfunction including seizures and psychosis. Its moderate form (15-50 microM) is associated with cognitive impairment and dementia. We investigated the neuropharmacological effects of homocysteine and its oxidized forms, homocysteinesulfinic acid (HCSA) and homocysteic acid (HCA), on neuronal network function utilizing dissociated cortical neurons from embryonic Wistar rats on microelectrode arrays. All substances inhibited dose-dependently and reversibly spontaneous neuronal network activity within seconds: L-HCSA and L-HCA blocked spontaneous spike rate (SSR) significantly at very low concentrations, with an IC50 of 1.9 and 1.3 microM, respectively; whereas the dose-response curve of D,L homocysteine revealed an IC50 of 401 microM. These effects were antagonized by 2 amino-5-phosphonovaleric acid (APV) pointing to the NMDA receptor as mediator of this fast and reversible inhibition of network activity. We conclude that a neuronal dysfunction observed in hyperhomocysteinemia is likely due to HCSA and HCA since effective concentrations of homocysteine are not reached in patients. PMID- 14759644 TI - Mathematical modelling of the use of macrophages as vehicles for drug delivery to hypoxic tumour sites. AB - Poor drug delivery and low rates of cell proliferation are two factors associated with hypoxia that diminish the efficacy of many chemotherapeutic drugs. Since macrophages are known to migrate specifically towards, and localize within, hypoxic tumour regions, a promising resolution to these problems involves genetically engineering macrophages to perform such anti-tumour functions as inducing cell lysis and inhibiting angiogenesis. In this paper we outline a modelling approach to characterize macrophage infiltration into early avascular solid tumours, and extensions to study the interaction of these cells with macrophages already present within the tumour. We investigate the role of chemotaxis and chemokine production, and the efficacy of macrophages as vehicles for drug delivery to hypoxic tumour sites. The model is based upon a growing avascular tumour spheroid, in which volume is filled by tumour cells, macrophages and extracellular material, and tumour cell proliferation and death is regulated by nutrient diffusion. Crucially, macrophages occupy volume, and hence contribute to the volume balance and hence the size of the tumour. We also include oxygen dependent production of macrophage chemokines, which can lead to accumulations in the hypoxic region of the tumour. We find that the macrophage chemotactic sensitivity is a key determinant of macrophage infiltration and tumour size. Although increased infiltration should be beneficial from the point of view of macrophage-based therapies, such infiltration in fact leads to increased tumour sizes. Finally, we include terms representing the induced death of tumour cells by hypoxic engineered macrophages. We demonstrate that reductions in tumour size can be achieved, but predict that a combination of therapies would be required for complete eradication. We also highlight some counter-intuitive predictions for example, absolute and relative measures of tumour burden lead to different conclusions about prognosis. In summary, this paper illustrates how mathematical models may be used to investigate promising macrophage-based therapies. PMID- 14759645 TI - Genetic codes as evolutionary filters: subtle differences in the structure of genetic codes result in significant differences in patterns of nucleotide substitution. AB - The codon-degeneracy model (CDM) predicts that patterns of nucleotide substitution in protein-coding genes are largely determined by the relative frequencies of four-fold (4f), two-fold, and non-degenerate sites, the attributes of which are determined by the structure of the governing genetic code. The CDM thus further predicts that genetic codes with alternative structures will "filter" molecular evolution differentially. A method, therefore, is presented by which the CDM may be applied to the unique structure of any genetic code. The mathematical relationship between the proportion of transitions at 4f degenerate nucleotide sites and the transition-to-transversion ratio is described. Predictions for five individual genetic codes, relative to the relationship between code structure and expected patterns of nucleotide substitution, are clearly defined. To test this "filter" hypothesis of genetic codes, simulated DNA sequence data sets were generated with a variety of input parameter values to estimate the relationship between patterns of nucleotide substitution and best fit estimates of transition bias at 4f degenerate sites for both the universal genetic code and the vertebrate mitochondrial genetic code. These analyses confirm the prediction of the CDM that, all else being equal, even small differences in the structure of alternative genetic codes may result in significant shifts in the overall pattern of nucleotide substitution. PMID- 14759646 TI - The role of trans-membrane signal transduction in turing-type cellular pattern formation. AB - The Turing mechanism (Phil. Trans. R. Soc. B 237 (1952) 37) for the production of a broken spatial symmetry in an initially homogeneous system of reacting and diffusing substances has attracted much interest as a potential model for certain aspects of morphogenesis (Models of Biological Pattern Formation, Academic Press, London, 1982; Nature 376 (1995) 765) such as pre-patterning in the embryo. The two features necessary for the formation of Turing patterns are short-range autocatalysis and long-range inhibition (Kybernetik 12 (1972) 30) which usually only occur when the diffusion rate of the inhibitor is significantly greater than that of the activator. This observation has sometimes been used to cast doubt on applicability of the Turing mechanism to cellular patterning since many messenger molecules that diffuse between cells do so at more-or-less similar rates. Here we show that Turing-type patterns will be able to robustly form under a wide variety of realistic physiological conditions though plausible mechanisms of intra cellular chemical communication without relying on differences in diffusion rates. In the mechanism we propose, reactions occur within cells. Signal transduction leads to the production of messenger molecules, which diffuse between cells at approximately equal rates, coupling the reactions occurring in different cells. These mechanisms also suggest how this process can be controlled in a rather precise way by the genetic machinery of the cell. PMID- 14759647 TI - Analysis of the effects of nitric oxide and oxygen on nitric oxide production by macrophages. AB - The interactions between NO and O(2) in activated macrophages were analysed by incorporating previous cell culture and enzyme kinetic results into a novel reaction-diffusion model for plate cultures. The kinetic factors considered were: (i) the effect of O(2) on NO production by inducible NO synthase (iNOS); (ii) the effect of NO on NO synthesis by iNOS; (iii) the effect of NO on respiratory and other O(2) consumption; and (iv) the effects of NO and O(2) on NO consumption by a possible NO dioxygenase (NOD). Published data obtained by varying the liquid depth in macrophage cultures provided a revealing test of the model, because varying the depth should perturb both the O(2) and the NO concentrations at the level of the cells. The model predicted that the rate of NO(2)(-) production should be nearly constant, and that the net rate of NO production should decline sharply with increases in liquid depth, in excellent agreement with the experimental findings. In further agreement with available results for macrophage cultures, the model predicted that net NO synthesis should be more sensitive to liquid depth than to the O(2) concentration in the headspace. The main reason for the decrease in NO production with increasing liquid depth was the modulation of NO synthesis by NO, with O(2) availability playing only a minor role. The model suggests that it is the ability of iNOS to consume NO, as well as to synthesize it, that creates very sensitive feedback control, setting an upper bound on the NO concentration of approximately 1 microM. The effect of NO consumption by other possible pathways (e.g., NOD) would be similar to that of iNOS, in that it would help limit net NO production. The O(2) utilized during enzymatic NO consumption is predicted to make the O(2) demands of activated macrophages much larger than those of unactivated ones (where iNOS is absent); this remains to be tested experimentally. PMID- 14759648 TI - Evolution of mutualism through spatial effects. AB - Mutualism among species is ubiquitous in natural ecosystems but its evolution is not well understood. We provided a simple lattice model to clarify the importance of spatial structure for the evolution of mutualism. We assumed reproductive rates of two species are modified through interaction between species and examine conditions where mutualists of both species, that give some benefit to the other species with their own cost, invade non-mutualists populations. When dispersal of offspring is unlimited, we verified the evolution of mutualism is impossible under any condition. On the other hand, when the dispersal is limited to neighboring lattice sites, mutualists can invade if the ratio of cost to benefit is low and the intrinsic reproductive rate is low in case where the parameter values are symmetric between species. Under the same conditions, non-mutualists cannot invade mutualist populations, that is, the latter are evolutionarily stable. In case of asymmetric parameters, mutualists tend to invade if the average value of costs to two species is low or that of benefits is high, and if the intrinsic reproductive rate is low for one of the two species. A mechanistic explanation of why mutualists increase when the dispersal is limited is given by showing that mutualist pairs of the two species at the same lattice site rapidly increase at the initial phase of the invasion. PMID- 14759649 TI - Why is it advantageous for animals to detect celestial polarization in the ultraviolet? Skylight polarization under clouds and canopies is strongest in the UV. AB - The perception of skylight polarization in the ultraviolet (UV) by many insect species for orientation purposes is rather surprising, because both the degree of linear polarization and the radiance of light from the clear sky are considerably lower in the UV than in the blue or green. In this work we call this the "UV-sky pol paradox". Although in the past, several attempts have been made to resolve this paradox, none of them was convincing. We present here a possible quantitative resolution to the paradox. We show by a model calculation that if the air layer between a cloud and a ground-based observer is partly sunlit, the degree of linear polarization p of skylight originating from the cloudy region is highest in the UV, because in this spectral range the unpolarized UV-deficient cloudlight dilutes least the polarized light scattered in the air beneath the cloud. Similarly, if the air under foliage is partly sunlit, p of downwelling light from the canopied region is maximal in the UV, because in this part of spectrum the unpolarized UV-deficient green canopylight dilutes least the polarized light scattered in the air beneath the canopy. Therefore, the detection of polarization of downwelling light under clouds or canopies is most advantageous in the UV, in which spectral range the risk is the smallest that the degree of polarization p is lower than the threshold p(tr) of polarization sensitivity in animals. On the other hand, under clear skies there is no favoured wavelength for perception of celestial polarization, because p of skylight is high enough (p > p(tr)) at all wavelengths. We show that there is an analogy between the detection of UV skylight polarization and the polarotactic water detection in the UV. However, insects perceive skylight polarization by UV or blue or green receptors. The question, why they differ in the spectral channel used for the detection of celestial polarization cannot be answered at the present time, because data are insufficient. Nevertheless, we present here one possible atmospheric optical reason why certain visual systems involved in detecting celestial polarization, are specifically tuned to the UV part of the spectrum. PMID- 14759650 TI - A three-dimensional vertex dynamics cell model of space-filling polyhedra simulating cell behavior in a cell aggregate. AB - We developed a three-dimensional (3D) cell model of a multicellular aggregate consisting of several polyhedral cells to investigate the deformation and rearrangement of cells under the influence of external forces. The polyhedral cells fill the space in the aggregate without gaps or overlaps, consist of contracting interfaces and maintain their volumes. The interfaces and volumes were expressed by 3D vertex coordinates. Vertex movements obey equations of motion that rearrange the cells to minimize total free energy, and undergo an elementary process that exchanges vertex pair connections when vertices approach each other. The total free energy includes the interface energy of cells and the compression or expansion energy of cells. Computer simulations provided the following results: An aggregate of cells becomes spherical to minimize individual cell surface areas; Polygonal interfaces of cells remain flat; Cells within the 3D cell aggregate can move and rearrange despite the absence of free space. We examined cell rearrangement to elucidate the viscoelastic properties of the aggregate, e.g. when an external force flattens a cell aggregate (e.g. under centrifugation) its component cells quickly flatten. Under a continuous external force, the cells slowly rearrange to recover their original shape although the cell aggregate remains flat. The deformation and rearrangement of individual cells is a two-step process with a time lag. Our results showed that morphological and viscoelastic properties of the cell aggregate with long relaxation time are based on component cells where minimization of interfacial energy of cells provides a motive force for cell movement. PMID- 14759651 TI - A logical analysis of the process of T cell activation: different consequences depending on the state of CD28 engagement. AB - The adaptive immune system is a complex organized action of several immune cell types like, T cells, B cells, dendritic cells, mast cells, and their ability to recognize self and foreign molecular information. Based on logical analysis, a model has been developed that describes TCR-ligand association coupled to intracellular signaling events that result in a proliferation signal. The model demonstrates that after TCR-ligand binding, the activation of tyrosine kinases in one of the paths leads to oscillations between the subsequent states of activation and deactivation of Ca(2+) initiation. In our studies the effect of costimulation on the primary signal has also been explored. Analysis reveals that costimulation increases by more than 2.5 fold the number of paths rendering a cell proliferation signal compared to the outcome when costimulation is blocked. Traversal of 97% of these paths attains a costimulation threshold of activation. We also examined a hypothesis that couples the primary signal and costimulation by modeling costimulation to act as an inhibitor on the Inhibitor proteins. Using this hypothesis our analysis showed a 25% increase in the number of paths leading to cell proliferation in comparison to when costimulation is blocked. Our model also reveals that this hypothesis actually decrease by approximately 50% the number of paths attaining cell proliferation compared to the number of available paths leading to cell proliferation when costimulation does not act as an inhibitor on Inhibitor proteins. This suggests that costimulation influences cell proliferation by providing a greater diversity of paths that converge to this state. However, costimulation should be thought independent of its regulatory interaction with the inhibitor proteins. PMID- 14759652 TI - Inbreeding coefficients for stochastically varying small population sizes-bias of calculation based on effective numbers. AB - The commonly used procedure to calculate inbreeding coefficients by effective population numbers (Ne) by the harmonic mean of generation-by-generation population sizes involves a computational bias. If the individual population sizes are considered as realizations of a binomially distributed random variable with sample size N and probability p, this bias can be investigated for the two cases p = constant and p = variable (Markov chain). The bias is of practical relevance only for small probabilities p, short period of initial successive generations, and small population sizes. The largest values for this computational bias are in the range of 0.05-0.06. It is concluded that for most practical purposes the approximate procedure is appropriate. PMID- 14759653 TI - A tiling approach to virus capsid assembly explaining a structural puzzle in virology. AB - A novel approach for the description of the protein stoichiometry of viral capsids, that is the protein shells protecting the viral genome, is introduced based on tiling theory. This approach generalizes Caspar-Klug theory of quasi equivalence to account also for non-quasi-equivalent subunit arrangements in icosahedral virus capsids that have been observed experimentally but are not covered by the Caspar-Klug approach. In particular, the new approach is used to explain the structure of polyoma virus, Simian Virus 40 and L-A virus capsids, which are considered structural puzzles in view of Caspar-Klug theory. PMID- 14759654 TI - Modeling threshold phenomena, metabolic pathways switches and signals in chemostat-cultivated cells: the Crabtree effect in Saccharomyces cerevisiae. AB - The long-term Crabtree effect in Saccharomyces cerevisiae cultivated in aerobic chemostat at steady state has been studied for three different substrate concentrations in the feed of the bioreactor (data: J. Gen. Microbiol., 129 (1983) 653). We have shown that a model using two ways of transport/metabolization (T/M) of hyperbolic form, with high and low affinity for the substrate, allowed to represent correctly the main characteristics of the phenomenon. The model is based on an explicit form of the T/M kinetics when the bioreactor is considered as a polyphasic dispersed system (PDS). Mass balances analysis also allows to quantify the critical dilution rate value (threshold), Dc, of the transition between respiratory and respirofermentative mode, for which ethanol is produced. A good approximation for the threshold is Dc = V(S)0 Y(Xc, S) where Y(Xc,S) is the average yield coefficient before transition and V(S)0, the maximum specific rate of high affinity T/M pathway. The theoretical value is 0.3 h(-1), and is equal to the experimental value. We thus show in a quantitative way that the transition depends both on culture conditions (global characteristic of the system) and on strain properties (intrinsic characteristic of the microorganism as well). Using two different methods to calculate the residual substrate has carried out the comparison between the simulations end the experimental data. This allowed showing that the latter is not well represented by Monod's model and has confirmed that the affinity for the substrate varies according to the biomass. We have then shown how to calculate the most important specific rates (or metabolic flux) related to biomass, ethanol, oxygen, hydrogen, respiratory and fermentative CO(2) and H(2)O within the cellular phase. It has appeared that the oxygen uptake rate directly depends on high-affinity T/M pathway. This let us think that the regulation of the Crabtree effect in S. cerevisiae depends on the saturation of some glucose metabolization and transport pathways rather than on saturation of the respiratory chains. The specific rates analysis has also allowed us to show, at least in this case, that the metabolization rate (biosynthesis+fueling) had its maximum value on the whole dilution rates interval; metabolites excretion (ethanol and fermentative CO(2)) only intervenes to drain a "surplus" glucose flux. As a consequence, the transport capacity must be higher than the one of metabolization. Maximization of the metabolization specific rate could then be used as an optimization criterion in the stoichiometric calculation of metabolic flux (and not the specific growth rate maximization because growth is limited in a chemostat (mu = D)). We have also shown that the mass balances based on the T/M processes are in agreement with molar and elementary balances of the general stoichiometric equation for glucose respiration and fermentation under aerobic conditions. Thanks to the specific rates calculating the stoichiometric coefficients has done this. The total mass balance difference does not exceed 4%, which is compatible with the experimental carbon balance. Finally, we have emphasized that the ratio of biosynthesis flux and metabolization flux is constant before and after transition. This observation could be applied as soon as the free substrate concentration in the cellular phase is low. The paper succinctly describes the former theoretical results on which the model is built and sufficiently explains the algorithm for straightforward implementation. PMID- 14759655 TI - Control of chemicals in Sweden: an example of misuse of the "precautionary principle". AB - With a background in biochemistry and radiation biology, I started to get involved in the control of chemicals area by battling the use of alkyl-mercury compounds in Swedish agriculture during the years 1964-1965 (C.-G. Rosen, H. Ackefors, and R. Nilsson, 1966, Seed dressing compounds based on organic mercury economic aspects and health hazards, Svensk Kemisk Tidskrift 78, 8-19), and subsequently I acted as the sole technical advisor to the plaintiffs for the thalidomide children in Sweden for 4 years, ending in a 100 million US dollar (present value) settlement with the producers in 1969 (H. Sjostrom and R. Nilsson, 1991, Thalidomide and the Power of the Drug Companies, Penguin, London, Feltrinelli, Milano, Iwanami Shoten, Tokyo Fisher Verlag, Berlin). I joined the Swedish EPA in 1974 and became head of the toxicological unit of the Products Control Division, where I was instrumental inter alia in pushing through regulations on reductions of lead in gasoline as well as the first general restrictions in world on the use of cadmium (R. Nilsson, 1989, Cadmium-an Analysis of Swedish Regulatory Experience, Report to the OECD Chemicals Group and Management Committee, January 1989). Since 1986 responsibility for control of chemicals was largely taken over from the Swedish EPA by the newly created National Chemicals Inspectorate (KEMI), an agency that employs me in the capacity of toxicologist. In between, I have been working for the OECD Chemicals Program as well as for WHO (IPCS) in various capacities and as a consultant in risk assessment for the US chemical industry under the Superfund Program. I was also associated with the Ministries of Environment of the governments of Iran and India. With respect to the latter, part of my recommendations were incorporated in the new Indian laws and regulations on chemicals that were issued subsequent to the Bhopal disaster (R. Nilsson, 1988, Procedures and Safeguards for Notification and Handling of Hazardous Chemicals in India, Report to WHO South East Asia Region, SEA/EH/391, April 1988). As a consequence of a decreasing role for science in regulatory affairs, and a corresponding increasing influence from politics, for the past 8 years I have devoted myself mainly to research-oriented activities in my capacity as adjunct professor of molecular toxicology and risk assessment at the Stockholm University. In international collaboration, my projects have been supported by the Directorate General for Science, Research and Development of the Commission of the European Communities, the US Chemical Manufacturers' Association, the Coulston Foundation (Alamogordo, State of New Mexico), L'Oreal, Merck Co., and in the past to a limited extent also by my own agency, the National Swedish Chemicals Inspectorate. However, my position as member of the executive board for the International Society of Regulatory Toxicology and Pharmacology in the United States, which I held for 5 years, as well as my role as technical advisor in products liability litigation, reflects my continued interest in the "politics of chemical risk." In this context my critical comments with respect to the current regulatory policy for control of chemicals have caused considerable concern within the Swedish regulatory establishment (R. Nilsson, M. Tasheva, and B. Jaeger, 1993. Why different regulatory decisions when the scientific information base is similar? I. Human risk assessment, Regul. Toxicol. Pharmacol. 17, 292-332; R. Nilsson, 1994, Problems in the regulation of carcinogenic chemicals in an international perspective. III. Critical assessment of regulatory approaches. Rev. Int. Contam. Ambiental, Mexico City, 10, 99-199; R. Nilsson, 1998, Integrating Sweden into the European Union: Problems concerning chemicals control. In The Politics of Chemical Risk-Scenarios for a Regulatory Future, R. Bal and Halffman, Eds, pp. 159-171, Kluwer Academic, Dordrecht). PMID- 14759656 TI - Recombinant Escherichia coli for the biomonitoring of benzene and its derivatives in the air. AB - For protection against environmental deterioration, pollutants should be reduced as much as possible. Therefore, a sensitive detection method for air pollutants is required, particularly for benzene, a compound with mutagenic, teratogenic, and carcinogenic properties. Some microorganisms have a number of enzymes that degrade organic compounds. The genetic information for many of these enzymes is codified in plasmids that usually comprise some elements such as proteins and RNA for their replication, and proteins for cell division control. Some plasmids have been obtained from Pseudomonas putida, a ubiquitous microorganism that is able to degrade biologic and inorganic compounds. P. putida contains the TOL plasmid, responsible for the degradation of benzene and its derivatives. The TOL plasmid has been fused with the gene for firefly luciferase (pTNS316 plasmid) to produce a luminescent bacterium (Escherichia coli HB101), which can be applied to the environmental monitoring of these chemicals. The recombinant E. coli transformed with the plasmid (E. coli HB101-pTNS316) was applied to the environmental biosensing of benzene and its derivatives. Measurement of bacteria-generated photons was done using a color-coded device to estimate quantitatively the pollutant concentration. The expression of luciferase was induced in the presence of aromatic compounds but the E. coli sensitivity has a detection limit: this bacterium dies if the benzene concentration is higher than 0.5 ppm and at this concentration the luminescence decreases so it is impossible to detect. Another limit for this biodevice is that the microorganism, very likely, accumulates the benzene and its derivatives, and therefore it is very important to consider the time of exposure. This biomonitor potentially offers a rapid, inexpensive, and sensitive technique for environmental detection of aromatic pollutant compounds. PMID- 14759657 TI - Sea urchin embryotoxicity test: proposal for a simplified bioassay. AB - Sea urchin embryotoxicity tests are widely used for evaluating the biological effects of contaminants in marine environments. The currently used traditional and standardized protocols are quite slow and laborious. The present work shows a modified bioassay (new embryotoxicity test; NET) in an attempt to speed up laboratory work using a limited number of fertilized eggs. Several experiments have been conducted both with a traditional bioassay and with the NET, using the same test conditions, in order to evaluate the reliability of the proposed simplified bioassay. Adult Paracentrotus lividus (Lamark) were collected from the Tyrrenian Sea (Bay of Naples) and embryos, reared in filtered seawater, were exposed to increasing potassium dichromate and copper sulfate concentrations. Then the EC(50) was calculated. The analysis of the results evidenced good repeatability. The confidence limits in all tests overlapped; moreover, data correlation analysis between the results of both tests showed a high significant accordance (chromium, R2 = 0.93, P < 0.01; copper, R2 = 0.86, P < 0/05). In conclusion, the NET seems to be a good alternative to the traditional tests; it could be a first step toward a new routine ecotoxicological kit for seawater. PMID- 14759658 TI - Effect of different toxic compounds on ATP content and acid phosphatase activity in axenic cultures of Tetrahymena pyriformis. AB - The sensitivity of protozoa, and particularly ciliated protozoa, to environmental changes suggested a study on the physiological responses arising from exposure to toxic compounds. Tetrahymena pyriformis was used as a test organism in a set of miniaturized assays. The physiological response of this ciliate was assessed in terms of adenosine-5'-triphosphate content and acid phosphatase activity after exposure of the cultures of T. pyriformis to four toxicants: copper, zinc, Triton X-100, and cycloheximide. In the range of concentrations used, stimulation and inhibition of these two parameters were observed. The correlation between the two parameters is analyzed. PMID- 14759659 TI - Effects of lead and exercise on endurance and learning in young herring gulls. AB - In this paper, we report the use of young herring gulls, Larus argentatus, to examine the effect of lead and exercise on endurance, performance, and learning on a treadmill. Eighty 1-day-old herring gull chicks were randomly assigned to either a control group or a lead treatment group that received a single dose of lead acetate solution (100mg/kg) at day 2. Controls were injected with an equal volume of isotonic saline at the same age. Half of the lead treatment group and half of the control group were randomly assigned to an exercise regime of walking on a treadmill twice each day. The other group remained in their cages. We test the null hypotheses that neither lead nor exercise affected performance of herring gull chicks when subsequently tested on the treadmill at 7, 11, and 17 days post-injection. Performance measures included latency to orient forward initially, to move continuously, forward on the treadmill, and to avoiding being bumped against the back of the test chamber. Also measured were the number of calls per 15 s, and the time to tire out. Latency to face forward and avoiding being bumped against the back of the test chamber were measures of learning, and time to tire out was a measure of endurance. We found significant differences as a function of lead, exercise, and their interaction, and rejected the null hypotheses. For all measures of behavior and endurance, lead had the greatest contribution to accounting for variability. In general, lead-treated birds showed better performance improvement from the daily exercise than did controlled non lead birds, with respect to endurance and learning. We suggest that in nature, exercise can improve performance of lead-exposed birds by partially mitigating the effects of lead, thereby increasing survival of lead-impaired chicks. PMID- 14759660 TI - Cold and drought stress in combination with pyrene exposure: studies with Protaphorura armata (Collembola: Onychiuridae). AB - Adult survival of the springtail Protaphorura armata exposed to pyrene, a common soil pollutant, was investigated in combination with cold and drought stress, in three separate experiments. (1) A drought stress imposed subsequent to pyrene exposure in soil resulted in a significant decrease in springtail survival, when compared with controls exposed to pyrene and subsequently to 100% relative humidity. (2) A previous exposure to drought stress resulted in slightly improved survival of pyrene exposure at a concentration of 10mg/kg, but not at higher pyrene concentrations. When comparing tests 1 and 2, better survival was found in the latter test. When comparing the drought survival of springtails that had been previously exposed to pyrene with drought survival of springtails with no previous history of pyrene exposure, survival was significantly lower in the former. (3) Springtail survival of pyrene exposure was investigated at several temperatures. Springtails showed a significant improvement in survival at temperatures fluctuating between +1 degrees C and -1 degrees C in 12:12-h cycles, and at a constant -3 degrees C, at the highest pyrene concentration (300 mg/kg), while survival remained the same at all temperatures when springtails were exposed to lower pyrene concentrations. It is concluded that temperature and water availability are important factors when assessing the springtails' susceptibility to pyrene exposure. PMID- 14759661 TI - Sensitivity of aquatic plants to the herbicide metsulfuron-methyl. AB - The sensitivity of 12 aquatic plant species to the herbicide metsulfuron-methyl was tested in microcosm experiments under two growth conditions. As reference species, barley (Hordeum vulgare) and oil-seed rape (Brassica napus) were grown with their roots submerged in the microcosms. Two response variables were chosen: relative growth rate and specific leaf area (SLA). SLA was the most sensitive response variable, with 11 of the 12 aquatic species responding to the herbicide. EC(50) values varied 56-fold between species, with the commonly used aquatic test species Lemna minor being one of the most sensitive. Fast-growing species with a small exposed leaf area proved to be more sensitive to the herbicide than slow growing species with a large exposed leaf area, which was believed to be primarily due to variations in growth rates rather than to variations in exposed leaf area. The aquatic plants displayed high tolerance in growth to metsulfuron compared with the sensitive crop oil-seed rape. Hence, possible spray-drift events and leaching of the herbicide applied at agricultural rates are not considered to have a large impact on the growth of the aquatic flora tested. PMID- 14759662 TI - Residual toxicity after biodegradation: interactions among benzene, toluene, and chloroform. AB - A microbial enrichment originating from a pristine aquifer was found to aerobically biodegrade benzene and toluene, but not chloroform. This enrichment culture was used to study changes in pollutant toxicity as affected by biodegradative activity. Two assays for toxicity were used: (1) a 48-h acute toxicity test using the freshwater invertebrate Ceriodaphnia dubia and (2) microbial biodegradation activity as affected by the presence of mixed pollutants. At 20-ppm concentrations, toluene was significantly more toxic (99% mortality) to C. dubia than benzene (48% mortality) or chloroform (40% mortality). Also at 20-ppm concentrations, but before biodegradation, toluene was significantly more toxic (88% mortality) to C. dubia than benzene (33% mortality). After biodegradation of 98% of toluene and benzene, significant residual toxicity still remained in the bacterial supernatant: toluene-degraded supernatant caused 33% mortality in C. dubia and benzene-degraded supernatant caused 24% mortality. In the second toxicity assay, examining the effect of mixed pollutants on biodegradation activity, the presence of benzene slowed the biodegradation of toluene, but chloroform had no effect on either benzene or toluene biodegradation. Results indicate that significant toxicity remain after biodegradation and that halogenated aliphatic hydrocarbons may have little or no effect on aromatic hydrocarbon biodegradation at sites impacted by mixed pollutants. PMID- 14759663 TI - Assessment of environmental stress by the micronucleus test and the Comet assay on the genome of teleost populations from two natural environments. AB - The aim of the present paper was to assess the biological damage caused by exposure of the test organism (Gambusia holbrooki: Cyprinodontiformes, Poecilidae) to various mutagenic agents present in the polluted waters of the Sarno River. For this purpose, we performed a micronuclei (MN) test and single cell gel electrophoresis (the Comet assay), testing DNA migration in an electrophoretic field using erythrocytes of G. holbrooki specimens both from the Sarno River and from the waters of the crater of the Astroni natural reserve as negative controls. The results indicate statistically higher values for both MN and DNA migration in the samples from the Sarno River compared with those from Astroni and point to a strong genotoxic action of the mixture of pollutants present in the Sarno River. These data were compared with the values found in the G. holbrooki specimens from the Sarno River kept under laboratory conditions for 100 days in clean water. PMID- 14759664 TI - Comparison of instantaneous rate of population increase and critical-effect estimates in Folsomia candida exposed to four toxicants. AB - The instantaneous rate of population increase (ri) integrates several life cycle variables into one accessible statistic and has been proposed as a more practical alternative than assembling full life tables in the study of population-level responses to toxicant exposure. In this study the sensitivity of instantaneous rate of population increase is compared to critical-effect estimates for populations exposed to four toxicants with different modes of action. Populations of the Collembolan Folsomia candida were exposed to cadmium, copper, pyrene, and chlorpyrifos in artificial soil following the standardized ISO (International Organization for Standardization, Geneva, 1999) protocol. We calculated ri values and LC(50), EC(50juvenile), and NOEC values for each chemical. Comparison of the relative toxicity of the four chemicals indicated that chlorpyrifos had the lowest values and was thus the most toxic, followed by pyrene, cadmium, and copper. Significant changes in ri were seen to follow closely changes in the sublethal parameter measured (juvenile production) and showed populations in decline at concentrations as low as 40% of the LC(50). The study showed ri to be a good measure of population response, and we conclude that the statistic gives a better understanding of effects on a population than through the sole use of traditional critical-effect estimates. PMID- 14759665 TI - Biochemical changes in the kidneys after perinatal intoxication with lead and/or cadmium and their antagonistic effects when coadministered. AB - Lead acetate (300 mg/L) and/or cadmium acetate (10 mg/L) were administered as drinking water to pregnant Wistar rats from day 1 of pregnancy to parturition (day 0) or until weaning (day 21) to investigate the possible nephrotoxic effects of these metals. We also studied the possibility of toxicological interactions between both metals. Kidneys were used to determine the activity of several enzymes considered key to correct renal function: alkaline and acid phosphatases, Mg(2+)/Ca(2+)-dependent ATPase, and Na(+)/K(+)-dependent ATPase. The results showed a general decrease in the activity of these enzymes after treatment with the heavy metals; this fact suggests that lead and cadmium are able to impair renal function due to biochemical alterations, since ATPases are essential for reabsorption and secretion processes and phosphatases are involved in the differentiation of the proximal tubules. On the other hand, simultaneous perinatal administration of both metals seems to protect against the toxicity produced by cadmium or lead separately. It is not clear whether this is due to decreased absorption or increased sequestration or excretion. PMID- 14759666 TI - Effects of pendimethalin at lower trophic levels--a review. AB - Pendimethalin's herbicidal action lies in its inhibition of the steps in plant cell division responsible for chromosome separation and cell wall formation. Terrestrial studies show that 10-20% of the herbicide evaporates during the first weeks after application. The remainder may dissipate biologically or chemically. Half-maximal dissipation time, or half-life (DT(50)) values vary from a few days to >200 days. Field and laboratory studies showed that reduced temperatures and drought prolong dissipation time to as long as 72-2094 days. In freshwater, pendimethalin concentrations reach 6 microg/L after runoff. In water the 10% lethal concentration (LC(10)) for Daphnia was 6 microg/L when exposed via suspended food. The LC(50) was 78 microg/L. An invertebrate field study showed that soil nematodes were reduced by 35-60% at pendimethalin application rates of 0.75 and 1.0 kg/ha, respectively. After application, soil microbiota is affected by pendimethalin for approximately 4 weeks. Plant-Rhizobium symbiosis is affected at application rates of 0.5-1.0 kg/ha. PMID- 14759667 TI - Role of piperonyl butoxide in the toxicity of chlorpyrifos to Ceriodaphnia dubia and Xenopus laevis. AB - The use of chemical inhibitors/inducers is one of the strategies employed to determine whether a particular metabolic pathway is involved in the metabolism of a xenobiotic. The objective of this study was to assess the role of piperonyl butoxide (PBO) on the toxicity of an organophosphorus insecticide, chlorpyrifos (CPF) to two species, Ceriodaphnia dubia (waterflea) and Xenopus laevis (South African clawed frog). Chlorpyrifos was highly toxic to C. dubia (48-h LC50: 0.05 microg/L) in comparison with X. laevis (96-h LC50: 2410 microg/L). Piperonyl butoxide at 200 microg/L reduced the toxicity of chlorpyrifos to C. dubia by a factor of 6. Piperonyl butoxide at 3000 microg/L also reduced the toxicity of CPF to X. laevis with respect to mortality and malformations. Acetylcholinesterase (AChE) activity was used as a biomarker to further assess the role of PBO in chlorpyrifos toxicity. X. laevis exposed to CPF and PBO exhibited a biphasic response in terms of AChE activity with an initial increase in the AChE activity followed by a drastic decrease. The results from the present study indicate that C. dubia and X. laevis have the capability to metabolize chlorpyrifos via cytochromes P450 mediated reactions. The results also indicate that the use of the biomarker AChE is useful in determining metabolic processes of organophosphorus insecticides, which require metabolic activation. PMID- 14759668 TI - Postexposure effects of DDE and butylbenzylphthalate on feeding behavior in threespine stickleback. AB - In a laboratory experiment we documented effects of sublethal concentrations of p,p'-2,2-bis(p-chlorophenyl)-1,1-dichloroethylene (DDE) and butylbenzylphthalate (BBP) on feeding behavior in threespine stickleback Gasterosteus aculeatus. The fish were exposed for 31 days to either BBP (10 or 100 microg/L) or DDE (5 or 50 microg/L) or to a mixture of BBP and DDE in the corresponding concentrations. Five weeks after exposure termination, we showed that fish that had been exposed to the higher concentrations of DDE and/or BBP initiated feeding more often than control fish. The latency time to feeding (ranging from 0.25 to 5.0 min) differed between control fish and fish exposed to mixtures of DDE and BBP. This experiment shows that feeding behavior may be used as a suitable behavioral variable in the detection of effects of pollutants even long time after the termination of exposure. PMID- 14759669 TI - Evaluation of the toxic effects of cypermethrin inhalation on the frog heart. AB - Cypermethrin is widely used as an insecticide on animals and in agriculture, the home, and the garden. The effect of inhaled cypermethrin on the cardiac mechanics, electrophysiology, and ultrastructure in frogs was investigated in this study. Four groups received 100 microL of cypermethrin via inhalation for different exposure times, and one group was used as a control. Electrical and mechanical activities of the heart were recorded, and heart samples were examined at light and transmission electron microscopic levels for all groups. The atrial and ventricular contractile forces on the mechanogram, the amplitude of the P wave and the QRS complex on electrocardiogram, and the heart rate were significantly decreased in cypermethrin-inhalated frogs. The total duration of contraction was prolonged in the study groups. Ultrastructurally, dilatation in smooth endoplasmic reticulum cisterns, a decrease in the number of mitochondria, disorganization in the myofibrils of myocytes, and necrotic changes in endothelial cells were observed. These results suggest that cypermethrin has cardiotoxic effects that increase with exposure time. PMID- 14759670 TI - Determination of aliphatic hydrocarbons in the alga Himanthalia elongata. AB - The algae considered new foods according to Regulation CE 258/97 need a guarantee of their healthfulness before being in the European market. In this work ten samples of the brown alga Himanthalia elongata have been analyzed with the aim of verifying the absence of aliphatic hydrocarbons, due to the ability of the macroalgae to capture lipophilic organic compounds of the marine water coming from accidental or continuous leaks of raw oil and refined products, which happen each year with the growth of the industrialization and the demand of energy. The fat of the samples were Soxhlet extracted using hexane:dichloromethane (1:1) for 7h. The organic fractions were purified using silica microcolumns. The identification and quantification of the aliphatic hydrocarbons have been carried out using gas chromatography (GC) with flame ionization detector (FID). The total hydrocarbon content was between 14.8 and 40.2 microg g(-1) dry weight. PMID- 14759671 TI - Economy and job contract as contexts of sickness absence practices: revisiting locality and habitus. AB - This study revisits two Finnish local governments-Raisio and Nokia-that in an earlier study showed different sickness absence rates in the early 1990s. The locality difference was interpreted sociologically, within a framework inspired by Bourdieu's theory of social field, habitus and practice. The same framework is applied in the present study, starting out from the hypothesis that a constant historical and cultural locality context tends to reproduce prevailing sickness absence practices. The hypothesis was tested by extending the context beyond the locality to the macroeconomic fluctuations that occurred during the 1990s and to the type of employment contract. In both localities a 30% rise was observed in levels of sickness absence from 1991-1993 to 1997-2000. At the beginning of the 1990s the absence rate among permanent employees was 1.86 times higher in Nokia than in Raisio; at the end of the decade the corresponding rate ratio was 1.88. The absence rates were significantly lower among fixed-term employees than permanent employees, but the locality difference was seen in their case, too. Both results support the hypothesis. In spite of major changes taking place in the national economy, the differences between the two towns' sickness absence rates persisted, which in this particular case probably reflects the persisting working-class character of Nokia and middle-class character of Raisio. The theory also applies to the difference between permanent and fixed-term employees: the peripheral power position of the latter on work related social fields leads to the observed practices, i.e. to the relatively low absence rate. The results of our revisit give reason to recapitulate and elaborate upon our theoretical interpretation with a view to deepening our understanding of the social origins of sickness absence practices in the post-industrial workplace, which is characterised by increasing atypical employment and growing job insecurity. PMID- 14759672 TI - Employee and union inputs into occupational health and safety measures in Chinese factories. AB - Few studies have addressed the impact of employees' inputs on the protection of their health and safety. The research presented in this paper focuses on Chinese factories and measures employees' evaluation of the effectiveness in OHS issues of their enterprise trade union and staff and workers' representative congress (SWRC). The data for the study draws upon a national survey of employees of enterprises in manufacturing industry conducted in 1997 by the All-China Federation of Trade Unions. The study finds that the input of the trade union and SWRC does have a significant impact on the protection of the workers' occupational health and safety. PMID- 14759674 TI - The impact of information on non-health attributes on willingness to pay for multiple health care programmes. AB - Despite the acceptance that health gain is the most important attribute of health care, other aspects of health care may affect utility. The aim of this paper is to report an experiment to test the impact of providing different levels of information in the context of the EuroWill study, a joint contingent valuation (CV) of multiple health programmes. Three hundred and three respondents were simultaneously asked for their willingness-to-pay (WTP) for three health care programmes: more heart operations, a new breast cancer treatment and a helicopter ambulance service. To test for the impact of variation in information, three versions of one of the programmes (heart) were provided. Results show that WTP for all three programmes tended to be significantly higher for respondents who were provided additional positive information about the heart programme. Our results show that CV of health care programmes, which only take into account medical outcomes, may lead to the value of such programmes not being adequately estimated, and that the impact of information may even be more decisive in the context of joint evaluation of multiple, rather than single, programmes. PMID- 14759673 TI - Employment status, social ties, and caregivers' mental health. AB - The purpose of this study of mid-life and older women was to assess the relation between informal care provision and depressive symptoms, taking into account concurrent demands on women's time (including multiple caregiving roles and employment outside the home) as well as participants' access to potentially supportive social ties. This cross-sectional study included women ages 46-71, free from major disease, who provided complete health and social information in the 1992 Nurses' Health Study follow-up survey (n = 61,383). In logistic regression models predicting depressive symptoms, we examined the interaction between employment outside the home and informal care provision for a disabled or ill spouse or parent. We also investigated level of social ties, measured with the Berkman-Syme Social Network Index, as a potential modifier of the association between informal care provision and depressive symptoms. In all analyses, higher weekly time commitment to informal care for a spouse or parent was associated with increased risk of depressive symptoms. This relationship persisted whether women were not employed outside the home, were employed full-time, or were employed part-time. Higher weekly time commitment to informal care provision was associated with increased risk of depressive symptoms whether women were socially integrated or socially isolated. However, both informal care provision and social ties were potent independent correlates of depressive symptoms. Therefore, women who reported high spousal care time commitment and few social ties experienced a dramatic elevation in depressive symptoms, compared to women with no spousal care responsibilities and many social ties (OR for depressive symptoms=11.8; 95% CI 4.8, 28.9). We observed the same pattern among socially isolated women who cared for their parent(s) many hours per week, but the association was not as strong (OR for depressive symptoms=6.5; 95% CI 3.4, 12.7). In this cross-sectional study, employment status did not seem to confer additional mental health risk or benefit to informal caregivers, while access to extensive social ties was associated with more favorable caregiver health outcomes. PMID- 14759675 TI - Spatial access to health care in Costa Rica and its equity: a GIS-based study. AB - This study assembles a geographic information system (GIS) to relate the 2000 census population (demand) with an inventory of health facilities (supply). It assesses the equity in access to health care by Costa Ricans and the impact on it by the ongoing reform of the health sector. It uses traditional measurements of access based on the distance to the closest facility and proposes a more comprehensive index of accessibility that results from the aggregation of all facilities weighted by their size, proximity, and characteristics of both the population and the facility. The weighting factors of this index were determined with an econometric analysis of clinic choice in a national household sample. Half Costa Ricans reside less than 1 km away from an outpatient care outlet and 5 km away from a hospital. In equity terms, 12-14% of population are underserved according to three indicators: having an outpatient outlet within 4 km, a hospital within 25 km, and less than 0.2 MD yearly hours per person. The data show substantial improvements in access (and equity) to outpatient care between 1994 and 2000. These improvements are linked to the health sector reform implemented since 1995. The share of the population whose access to outpatient health care (density indicator) was inequitable declined from 30% to 22% in pioneering areas where reform began in 1995-96. By contrast, in areas where reform has not occurred by 2001, the proportion underserved has slightly increased from 7% to 9%. Similar results come from a simpler index based on the distance to the nearest facility. Access to hospital care has held steady in this period. The reform achieved this result by targeting the least privileged population first, and by including such measures as new community medical offices and Basic Teams for Integrated Health Care (EBAIS) to work with these populations. The GIS platform developed for this study allows pinpointing communities with inadequate access to health care, where interventions to improve access would have the greatest impact. PMID- 14759677 TI - Lay voices on allergic conditions in children: parents' narratives and the negotiation of a diagnosis. AB - Allergic conditions can be seen as an increasing as well as debated health problem in Western societies, but lay notions and experiences of these conditions are still not fully understood. As much attention is given to prevention of allergic conditions in early childhood, for example as medical advice to parents of young children, it is of particular interest to look at lay understandings of allergic conditions in childhood. This study, carried out in Sweden, explores understandings of child allergy, drawing on interviews with parents of children under 6 years, in a period when the children are medically assessed. The interviews are analysed as illness narratives, with a focus on how the parents explain the child's illness. The analysis reveals a complex pattern. The parents on the one hand refer to a shared knowledge about causes to allergic conditions, such as factors in the physical environment, family life-style and genetic causes. On the other hand, this knowledge is re-appropriated and intertwined with the parents' own experiences of allergic conditions in the process of making sense of the illness in their own child. In their stories, the parents link a potential allergic condition in the child to their own identities as allergic or non-allergic persons and to their family illness history. Child allergy is in this sense constructed as a "family condition". PMID- 14759676 TI - Emergency contraception and morality: reflections of health care workers and clients. AB - In this study, we explore the retrospective reports of 21 US Planned Parenthood clients about their use of emergency contraception pills (ECPs) and the views of ten Planned Parenthood health care workers at two clinics about providing ECPs. We elucidate the sociological phenomena that frame emergency contraception usage: cultural ideology about contraception, sexuality, unintended pregnancy, and abortion. We focus on the ways in which interactions between health care workers and clients both mediate and reinforce such cultural ideology. Our research indicates that the distinctions between fertilization and pregnancy, between contraception and abortion, between responsible and irresponsible procreative behavior, are not hard and fast boundaries upon which everyone agrees. We illuminate the dividing lines and continuities our participants invoked, affirmed, and questioned when contemplating the continuum from potential fertility to realized (and unwanted) pregnancy. PMID- 14759678 TI - Health literacy, social support, and health: a research agenda. AB - Amid increased concerns about the adverse consequences of low health literacy, it remains unclear how health literacy affects health status and health service utilization. Moreover, studies have shown significant variation in individual adaptation to health literacy problems. This article proposes research hypotheses to address two questions: (1) What are the causal pathways or intermediate steps that link low health literacy to poor health status and high utilization of expensive services such as hospitalization and emergency care? (2) What impact does social support have on the relationships between health literacy and health service utilization? Empirical studies of health literacy are reviewed to indicate the limitations of current literature and to highlight the importance of the proposed research agenda. In particular, we note the individualistic premise of current literature in which individuals are treated as isolated and passive actors. Thus, low health literacy is considered simply as an individual trait independent of support and resources in an individual's social environment. To remedy this, research needs to take into account social support that people can draw on when problems arise due to their health literacy limitations. Examination of the proposed agenda will make two main contributions. First, we will gain a better understanding of the causal effects of health literacy and identify missing links in the delivery of care for patients with low health literacy. Second, if social support buffers the adverse effects of low health literacy, more effective interventions can be designed to address differences in individuals' social support system in addition to individual differences in reading and comprehension. More targeted and more cost-efficient efforts could also be taken to identify and reach those who not only have low health literacy but also lack the resources and support to bridge the unmet literacy demands of their health conditions. PMID- 14759679 TI - From symptom recognition to diagnosis: children with autism in urban India. AB - The period of time between initial recognition of a symptom and initial diagnosis warrants examination because it can serve as a window into broader cultural factors and allow for more immediate treatment. Research on this process among parents of autistic children to date has been useful, but has been limited to families in Western countries, whereas autism occurs all over the world. The current study provides data on the process of initial symptom recognition, help seeking, and initial diagnosis of a pervasive developmental disorder among 95 families of autistic children in India. The findings suggest that the specific symptoms that parents initially recognize may be associated with the speed with which they receive a diagnosis, and that the saliency of symptoms may be culturally shaped. Environmental, cultural, and socioeconomic factors may also have an impact on whether a parent receives a diagnosis. Implications for professional awareness and increased accuracy of diagnosis are also suggested. PMID- 14759680 TI - School-based HIV prevention programmes for African youth. AB - The high rate of HIV infection among youth in Africa has prompted both national and international attention. Education and prevention programmes are seen as the primary way of decreasing this rate. This paper reviews 11 published and evaluated school-based HIV/AIDS risk reduction programmes for youth in Africa. Most evaluations were quasi-experimental designs with pre-post test assessments. The programme objectives varied, with some targeting only knowledge, others attitudes, and others behaviour change. Ten of the 11 studies that assessed knowledge reported significant improvements. All seven that assessed attitudes reported some degree of change toward an increase in attitudes favourable to risk reduction. In one of the three studies that targeted sexual behaviours, sexual debut was delayed, and the number of sexual partners decreased. In one of the two that targeted condom use, condom use behaviours improved. The results of this review suggest that knowledge and attitudes are easiest to change, but behaviours are much more challenging. The article provides details about programmes and identifies characteristics of the most successful programmes. Clearly, however, more research is needed to identify, with certainty, the factors that drive successful school-based HIV/AIDS risk reduction programmes in Africa. PMID- 14759681 TI - Social support and quality of life over time among adults living with HIV in the HAART era. AB - Stability in perceived social support and associations between social support and health-related quality of life for a sample of 41 adult outpatients living with HIV/AIDS (PHA) in Canada were assessed longitudinally. Construct-specific dimensions of the Medical Outcomes Study Social Support Survey (SSS), the Physical and Mental components of the Short-Form-36 (SF-36) quality of life measure, as well as clinical factors (i.e., symptomatology, immunologic/virologic variables), were measured in three waves: initial consecutive registration (T1, 1997), 2-year (T2, 1999) and 4-year (T3, 2001) follow-up, and evaluated for changes using repeated-measures analysis of variance, supplemented by Friedman tests for SSS and SF-36 ratings. Proportions of the PHA sample with clinically significant SSS changes (i.e., greater than 0.5 standardized effect size) were also calculated. Effects of improvement versus deterioration in SSS ratings on SF 36 ratings, and vice versa, were explored. Associations between SSS and SF-36 ratings, as well as between changes in SSS ratings and SF-36 ratings, were assessed using multiple regression analyses controlling for clinical factors. Cross-lagged analyses were conducted to examine predictive potential between SSS and SF-36 ratings. Clinical outcomes suggested immunologic improvement tempered by symptoms and/or treatment side effects. SSS and SF-36 mean ratings were moderately stable over time, but clinically significant 4-year decrements in SSS ratings occurred for approximately 40% of patients. A trend occurred in which poorer SF-36 mental outcomes portended poorer emotional and informational support. Otherwise, relations between SSS and SF-36 ratings appeared to be reciprocal. Cross-sectional associations between SSS and SF-36 ratings were more pronounced at T2 compared to baseline and T3. Changes in SSS and SF-36 ratings were somewhat related over the consecutive 2-year periods but not over the long term. T1-T2 SSS changes were associated with changes in the SF-36 mental component. T2-T3 SSS changes were associated with changes in the SF-36 physical component. Cross-lagged analyses yielded little explanation concerning direction of causation in terms of associations between social support and quality of life for the PHA in this study. PMID- 14759682 TI - Effective/efficient mental health programs for school-age children: a synthesis of reviews. AB - The prevalence of mental health problems, some of which seem to be occurring among younger cohorts, leads researchers and policy-makers to search for practical solutions to reduce the burden of suffering on children and their families, and the costs to society both immediate and long term. Numerous programs are in place to reduce or alleviate problem behaviour or disorders and/or assist positive youth development. Evaluated results are dispersed throughout the literature. To assess findings and determine common elements of effective children's services, a literature search was undertaken for evidence based evaluations of non-clinical programs for school-age children. Prescriptive comments aim to inform service-providers, policy-makers and families about best practices for effective services such as: early, long-term intervention including reinforcement, follow-up and an ecological focus with family and community sector involvement; consistent adult staffing; and interactive, non-didactic programming adapted to gender, age and cultural needs. Gaps are identified in our understanding of efficiencies that result from effective programs. Policy implications include the need to develop strategies for intersectoral interventions, including: new financing arrangements to encourage (not penalize) interagency cooperation and, to ensure services reach appropriate segments of the population; replication of best practices; and publicizing information about benefits and cost savings. In many jurisdictions legislative changes could create incentives for services to collaborate on service delivery. Joint decision-making would require intersectoral governance, pooling of some funding, and policy changes to retain savings at the local level. Savings could finance expansion of services for additional youth. PMID- 14759683 TI - Family consequences of chronic back pain. AB - This paper considers occupational back injuries as they are described by injured workers, including the impact on their abilities to function in family and social roles. In this study, we interviewed people with work-related back injuries in Florida and Wisconsin. The sample was chosen from among workers who either lost more than 28 days from work or who received workers' compensation permanent partial disability benefits. Respondents indicated a wide range of limitations on family and social roles, including physical impacts that hampered respondents' ability to do household chores, to take part in raising children, and to engage in leisure activities with their spouses. Spouses and children took over family responsibilities once carried on by injured workers. The impacts of limitations resulted in a restructuring of family and social roles, relationships, and self identities. In addition, these impacts led to depression and anger among the injured workers and to stress and strain in family relationships. Unlike the effects on work, many of these outcomes are not valued in the marketplace and, as a consequence, are less visible. They are nonetheless important and suggest that priorities based only on economic outcomes may understate the importance of preventive activities. Moreover, they suggest that attention should be focused on social structural factors such as the provision of medical services and social support to reduce the impact of chronic back pain on family members. PMID- 14759684 TI - Caregiver's needs as predictors of hospital readmission for the elderly in Taiwan. AB - The problems of caregivers are identified as one of the major causes, as well as a preventable one, for the unplanned hospital readmissions of elderly patients. Family caregivers are found to be under intense stress and burden, and to encounter many difficulties preparing themselves, both physically and mentally, for the return home from the hospital of an elderly family member. However, few studies of rehospitalization of the elderly have focused on the point of view of the family caregiver. The purpose of this study was to explore the predictors, especially the needs of the family caregiver, for rehospitalization, within a month after having been discharged, for elderly patients in Taiwan. A prospective, descriptive and correlational study, using 216 elderly patients and their family caregivers from the internal medicine wards of a medical center in Northern Taiwan was conducted. It was found that elderly patients with caregivers requesting help from a support group were 5.3 times more likely to be rehospitalized than those with caregivers that did not request such help. In addition, the odds of rehospitalization for elderly patients with some self-care ability, whose caregiver reported at least one need for social services, was 9.03 (CI=2.32-35.16) times that of those whose caregivers reported no need for social services. Based on the findings of this study, health care providers need to assess the self-care ability of their elderly patients. In addition they should assess the need for support groups and social services to caregivers before the hospital discharge of Taiwanese elders, and perhaps other Chinese populations as well. PMID- 14759685 TI - Parental perspectives on vaccinating children against sexually transmitted infections. AB - Several vaccines for sexually transmitted infections (STI) are presently in development and the eventual availability of such vaccines is expected to result in the prevention of a significant number of burdensome conditions. Young adolescents are presumed to be likely targets for these vaccines since adolescents' risk for STI increases as they age and become sexually active. It is unclear, however, to what extent parents will agree to having adolescents receive STI vaccines. Inasmuch as acceptance is the foundation for effective immunization programs, an understanding of parental perspectives about this issue is required to inform future STI vaccine program strategies. This paper presents findings from a qualitative study that used in-depth interviews to elicit attitudes from 34 parents about accepting vaccines for genital herpes, human immunodeficiency virus, human papillomavirus and gonorrhea for their children (aged 8-17). Data were collected from parents bringing their children for care at an urban clinic and a suburban private office. Content analysis of the responses revealed that most parents (>70%) approved the administration of all four of the STI vaccines proposed. Parents' reasons for acceptance included wanting to protect their children, being concerned about specific disease characteristics, and previous experience with the infections. Parents who declined the vaccines did so primarily because they perceived their children to be at low risk for the infections or they had low concern about features of the diseases. Most parents thought they should be the decision-maker regarding children receiving an STI vaccine. Results from this study will be used to plan subsequent investigations of the determinants of STI vaccine acceptance by parents. PMID- 14759686 TI - Non-professionals' evaluations of gene therapy ethics. AB - Although the moral responsibilities of clinicians and researchers in the new genetics are exhaustively reflected upon, much less attention has been paid to the factors affecting the moral reasoning of non-professionals when they reflect on genetic issues. In this paper, we compare the moral evaluations of somatic gene therapy (SGT) made by some of its potential consumers (patients) and its providers (medical professionals). The results highlight significant differences between professional opinion and non-professional evaluations. Medical professionals shared a moral evaluation of SGT that (a) based its acceptability on a strong therapeutic imperative, (b) grounded this in an unproblematic separation of identity and disability/illness, and (c) generally did not see SGT as ethically different from other medical interventions. Prospective patients (a) often questioned the effectiveness of "therapeutic" interventions, (b) could derive a strong sense of identity from disability/illness, and (c) sometimes saw genetic interventions as changing a person's identity, either directly (through the genes) or indirectly (through altered life experience). We discuss the implications of these differences for the professional and public debate on the ethics of gene therapy. Our results highlight the need to take into account non professionals' views of SGT. PMID- 14759687 TI - Assessment of genetic testing and related counseling services: current research and future directions. AB - With the recent completion of the sequencing of the Human Genome, genetic testing will increasingly become available for a greater number of medical conditions, many of which are those that manifest in adulthood (e.g., various cancers, cardiovascular disease, diabetes) or for which little or no treatments are available (e.g., Alzheimer disease). Genetic services, defined here as those relating to genetic testing and counseling, will be with helping more individuals deal with medical information that affects their health directly, as opposed to affecting primarily the health of their offspring. This paper reviews the existing research in the genetic testing and counseling literature and presents an evaluation framework outlining the intended outcomes of genetic services. The purpose of this framework is to provide an overview of the potential outcomes of these services and highlight constructs for future research in this area. In addition, other issues that will affect the assessment of genetic services are raised, using examples from the existing literature. Ultimately, the goal of this paper is to highlight and suggest directions researchers can take to produce the information needed to guide genetic testing and counseling practice. Moreover, as genetic knowledge is increasingly applied towards the prevention and treatment of various common, chronic disease conditions, genetic information will have implications for providers outside of the traditional medical genetics realm, such as primary care providers and public health practitioners. A better understanding of the outcomes of genetic testing and counseling will provide a basis from which to ensure an appropriate application of genetic information by all those who eventually provide care and "genetic" services. PMID- 14759688 TI - Fosterage patterns in the age of AIDS: continuity and change. AB - An estimated 4 million children, or about 10% of the entire South African population, will be orphaned by the year 2015. There is growing consensus that the extended family system is no longer capable of providing for orphans given severe economic constraints. There is, therefore, an urgency to develop appropriate interventions to support families and take care of these children. This article examines some of the existing literature on child fosterage and uses it to highlight understudied aspects of the current situation of children orphaned through AIDS in South Africa. Of particular concern are the points of continuity and change in fosterage patterns before and after the onset of the epidemic in South Africa. I suggest that an understanding of the short- and long term consequences for children orphaned by AIDS in South Africa calls for historical contextualisation given that child fostering, both voluntarily and involuntarily, has been a feature of black family life since well before the onset of HIV/AIDS. In addition, I demonstrate the value of examining kinship, family, and networks in order to fully understand the circumstances of fostering these children. The paper concludes with a call for more research on children orphaned by AIDS in South Africa that will provide not only more data, but also enrich theoretical approaches to studying patterns of child fosterage in Africa and elsewhere. PMID- 14759690 TI - Health inequalities and the psychosocial environment-two scientific challenges. AB - As social inequalities in health continue to be a key public health problem, scientific advances in explaining these inequalities are needed. It is unlikely that there will be a single explanation of social inequalities in health. This introductory paper sets out one explanatory framework, exposure to adverse psychosocial environments during midlife, and particularly at work. We argue that exposure to an adverse psychosocial environment, in terms of job tasks, defined by high demands and low control and/or by effort-reward imbalance, elicits sustained stress reactions with negative long-term consequences for health. These exposures may be implicated in the association of socioeconomic status with health in two ways. First, these exposures are likely to be experienced more frequently among lower socioeconomic groups. Second, the size of the effects on health produced by adverse working conditions may be higher in lower status groups, due to their increased vulnerability. In this special issue, these arguments are illustrated by a collection of original contributions from collaborative research across Europe. The papers, in our view, advance the case for the robust associations between measures of adverse psychosocial environment and ill health, as they are based on comparative studies across several European countries and as they combine different types of study designs. This collaboration was enabled and supported by a European Science Foundation scientific programme on 'Social Variations in Health Expectancy in Europe'. PMID- 14759691 TI - Psychosocial factors at work and depression in three countries of Central and Eastern Europe. AB - Psychosocial factors at work have been found to predict a range of health outcomes but their effect on mental health outcomes has not been extensively studied. This paper explores the relationship between psychosocial factors at work and depression in three countries of Central and Eastern Europe. The data come from a cross-sectional study of working men (n = 645) and women (n = 523) aged 45-64 years, randomly selected from population registers in Novosibirsk (Russia), Krakow (Poland) and Karvina-Havirov (Czech Republic). The questionnaire included questions on the effort and reward at work, job control, the full CES-D scale of depression, and a range of other characteristics. Linear regression was used to estimate the association between depression score and work characteristics: the logarithm of the effort-reward ratio, and continuous job control score. The means of the depression score were 10.5 for men and 14.2 for women. After controlling for age, sex and country, effort-reward ratio (logarithmically transformed) was strongly related to depression score; a 1 SD increase in the log transformed effort-reward ratio was associated with an increase in the depression of 2.0 points (95% CI 1.5; 2.4), and further adjustment did not materially change the effect. Job control was inversely associated with depression score in Poland and the Czech Republic (not in Russia) but the association was largely eliminated by controlling for socioeconomic characteristics. This study suggests that the effort-reward imbalance at work is related to prevalence of depression in these central and eastern European populations. PMID- 14759692 TI - The measurement of effort-reward imbalance at work: European comparisons. AB - Using comparative data from five countries, this study investigates the psychometric properties of the effort-reward imbalance (ERI) at work model. In this model, chronic work-related stress is identified as non-reciprocity or imbalance between high efforts spent and low rewards received. Health-adverse effects of this imbalance were documented in several prospective and cross sectional investigations. The internal consistency, discriminant validity and factorial structure of 'effort', 'reward', and 'overcommitment' scales are evaluated, using confirmatory factor analysis. Moreover, content (or external) validity is explored with respect to a measure of self-reported health. Data for the analysis is derived from epidemiologic studies conducted in five European countries: the Somstress Study (Belgium; n = 3796), the GAZEL-Cohort Study (France; n = 10,174), the WOLF-Norrland Study (Sweden; n = 960), the Whitehall II Study (UK; n = 3697) and the Public Transport Employees Study (Germany; n = 316). Internal consistency of the scales was satisfactory in all samples, and the factorial structure of the scales was consistently confirmed (all goodness of fit measures were > 0.92). Moreover, in 12 of 14 analyses, significantly elevated odds ratios of poor health were observed in employees scoring high on the ERI scales. In conclusion, a psychometrically well-justified measure of work-related stress (ERI) grounded in sociological theory is available for comparative socioepidemiologic investigations. In the light of the importance of work for adult health such investigations are crucial in advanced societies within and beyond Europe. PMID- 14759693 TI - The effect of control at home on CHD events in the Whitehall II study: Gender differences in psychosocial domestic pathways to social inequalities in CHD. AB - Although there has been considerable research on psychosocial working conditions and their effect on physical and mental health, there has been little research into the effects of psychosocial domestic conditions on health. The association between psychosocial working conditions (and control at work in particular) and coronary heart disease (CHD) is not as strong for women compared to men. Other research suggests that household and domestic factors may have an important effect on women's health. Some studies have shown that low control at home affects psychological well being. However, there has been little research into its effects on physical health. Furthermore, similar to results analysing low control at work, low control at home may form part of the pathways underlying social inequalities in health. The study investigates the meaning of control at home, the effect of control at home on incident CHD events and whether this explains some of the social inequalities in CHD events in men and women. Data from phases 3-5 of the Whitehall II study, London, UK, were analysed (N = 7470). The results indicate that low control at home predicts CHD among women but not among men. Furthermore, low control at home may explain part of the association between household social position and CHD among women. There is some evidence suggesting that low control at home among women results from a lack of material and psychological resources to cope with excessive household and family demands. Psychosocial domestic conditions may have a greater effect on the health of women compared with men. PMID- 14759694 TI - Psychobiological mechanisms of socioeconomic differences in health. AB - The association between low socioeconomic status and poor health is well established. Empirical studies suggest that psychosocial factors are important mediators for these effects, and that the effects are mediated by psychobiological mechanisms related to stress physiology. The objective of this paper is to explore these psychobiological mechanisms. Psychobiological responses to environmental challenges depend on acquired expectancies (learning) of the relations between responses and stimuli. The stress response occurs whenever an individual is faced with a challenge. It is an essential element in the total adaptive system of the body, and necessary for adaptation, performance and survival. However, a period of recovery is necessary to rebalance and to manage new demands. Individuals with low social status report more environmental challenges and less psychosocial resources. This may lead to vicious circles of learning to expect negative outcomes, loss of coping ability, strain, hopelessness and chronic stress. This type of learning may interfere with the recovery processes, leading to sustained psychobiological activation and loss of dynamic capacity to respond to new challenges. Psychobiological responses and health effects in humans and animals depend on combinations of demands and expected outcomes (coping, control). In studies of humans with chronic psychosocial stress, and low SES, cortisol baseline levels were raised, and the cortisol response to acute stress attenuated. Low job control was associated with insufficient recovery of catecholamines and cortisol, and a range of negative health effects. Biological effects of choice of lifestyle, which also depends on the acquired outcome expectancies, reinforce these direct psychobiological effects on health. The paper concludes that sustained activation and loss of capacity to respond to a novel stressor could be a cause of the higher risk of illness and disease found among people with lower SES. PMID- 14759695 TI - Work stress, socioeconomic status and neuroendocrine activation over the working day. AB - Socioeconomic status (SES) differences in cardiovascular and metabolic disease risk may be mediated in part by differential activation of neuroendocrine pathways. We have previously found that salivary cortisol levels over the working day are greater in lower than higher SES men, but that cortisol output is greater in higher than lower SES women. This study investigated the role of work stress in generating these patterns, analysing cortisol output in relation to job demands and job control. Participants were 97 men and 84 woman from the Whitehall II cohort, London, UK, recruited from higher and lower grades of employment. Saliva samples were obtained on waking and 30 min later to assess the cortisol waking responses, and at two hourly intervals over a typical working day. Cortisol responses to waking were positively associated with high job demands, but this effect was attenuated by higher SES. In women but not men, cortisol levels over the remainder of the day were elevated in lower SES participants who experienced high job demands, but depressed in lower status women who reported low job demands. Job control did not influence cortisol responses to waking, but in men cortisol levels over the remainder of the day were inversely related to job control. These cortisol differences were independent of age, smoking status and time of waking up. Subjectively, the most stress was reported by higher SES individuals who experienced low job control. We conclude that work stress and SES are related differently to cortisol responses to waking and cortisol output over the day. Job control may partly mediate SES differences in cortisol in men, while job demands are more relevant for women. Analyses of psychobiological pathways must take account of variations in exposure to chronic stressors as well as differences in responsivity to stressors. PMID- 14759696 TI - Effort-reward imbalance model and self-reported health: cross-sectional and prospective findings from the GAZEL cohort. AB - Psychosocial factors at work have been found to be significant contributors to health, especially cardiovascular health. This study is aimed at exploring the relationship between psychosocial factors at work as defined by the effort-reward imbalance (ERI) model and self-reported health, using alternative formulations of this model, and comparing cross-sectional and prospective analyses for a large occupational cohort of men and women. The French version of the ERI model was used to measure the three scales of effort, reward, and overcommitment. Self reported health was used as health outcome. Covariates included chronic diseases, frequent depressive symptoms, and personal, occupational, and behavioural factors. The cross-sectional and prospective analyses concerned, respectively, 10175 and 6286 workers. Men and women were analysed separately. Cross-sectional analysis revealed that ERI was significantly associated with self-reported health whatever the formulation used (ratio over one, quartiles, continuous ratio, or log-transformed ratio) for both genders. When effort and reward were studied as two separate variables, reward was a significant risk factor for both genders, whereas effort was for men only. Overcommitment was also found to be a risk factor for self-reported health for both sex. Prospective analysis showed that ERI was a significant predictor of poor self-reported health for men and women for two formulations (continuous ratio and log-transformed ratio). For both genders, effort did not predict self-reported health, but reward did. Overcommitment was predictive of poor self-reported health for men only. Our results highlighted the predictive effects of the ERI model on self-reported health in a 1-year follow-up study. They urged to explore various formulations of the ERI model. They also underlined the need for longitudinal study design and separate analyses for men and women in the field of psychosocial factors at work. PMID- 14759697 TI - Differential economic stability and psychosocial stress at work: associations with psychosomatic complaints and absenteeism. AB - Stressful working conditions are well known to have a negative impact on the worker's health. We investigated this association in a Belgian study with a psychosocial health perspective, including individual work characteristics as well as firms' features. These data come from the first measure of the Somstress study. This is a 4 year project, initiated in 1999 and conducted in four different firms. The objective of this article is to investigate the relationships between stress, working conditions and absenteeism, self-reported health and psychosomatic complaints. Firms were selected according to their degree of structural environment and job stability. Among the four work sites, one can be considered as stable, one unstable and the remaining ones in an in between situation. Stress is generally measured according to one of the following models: the job demands control model (Karasek) and the effort-reward imbalance model (Siegrist). We used here both models, along with the social support at work (Karasek) and overcommitment (Siegrist). Sex, age and education are important health determinants. After adjustment for those three variables and additionally for the work instability, it appeared that poor health outcomes (measured by the self-rated health, depression (SCL-90), anxiety (SCL-90), somatisation (SCL-90), chronic fatigue (Vercoulen) and reported absenteeism) are mainly associated with a low control, low social support at work, high overcommitment and high level of imbalance. Inversely, job demands do not make any significant contribution in the logistic regression models for the above-mentioned health outcomes. PMID- 14759698 TI - Can we disentangle life course processes of accumulation, critical period and social mobility? An analysis of disadvantaged socio-economic positions and myocardial infarction in the Stockholm Heart Epidemiology Program. AB - The accumulation hypothesis would propose that the longer the duration of exposure to disadvantaged socio-economic position, the greater the risk of myocardial infarction. However there may be a danger of confounding between accumulation and possibly more complex combinations of critical periods of exposure and social mobility. The objective of this paper is to investigate the possibility of distinguishing between these alternatives. We used a population based case-control study (Stockholm Heart Epidemiology Programme) of all incident first events of myocardial infarction among men and women, living in the Stockholm region 1992-94. The analyses were restricted to men 53-70 years, 511 cases and 716 controls. From a full occupational history each subject was categorized as manual worker or non-manual at three stages of the life course, childhood (from parent's occupation), at the ages 25-29 and 51-55, resulting in 8 possible socio-economic trajectories. We found a graded response to the accumulation of disadvantaged socio-economic positions over the life course. However, we also found evidence for effects of critical periods and of social mobility. A conceptual analysis showed that there are, for theoretical reasons, only a limited number of trajectories available, too small to form distinct empirical categories of each hypothesis. The empirical task of disentangling the life course hypotheses of critical period, social mobility and accumulation is therefore comparable to the problem of separating age, period, and cohort effects. Accordingly, the interpretation must depend on prior knowledge of more specific causal mechanisms. PMID- 14759699 TI - Social mobility and health in the Turin longitudinal study. AB - One of the most controversial explanations of class inequalities in health is the health selection hypothesis or drift hypothesis which suggests there is a casual link between the health status of individuals and their chances of social mobility, both inter- and intra-generational. This study tests this hypothesis, and tries to answer three related questions: (a) to what extent does health status influence the chances of intra-generational mobility of individuals? (b) what is the impact on health inequalities of the various kinds of social mobility (both mobility in the labour market and exit from employment)-do they increase or reduce inequalities? (c) to what extent does health-related intra-generational social mobility contribute to the production of health inequalities? The data analysed in this paper were drawn from the records of the Turin Longitudinal Study, which was set up to monitor health inequality of the Turin population by combining census data, population registry records and medical records. Occupational mobility was observed during the decade 1981-1991. To evaluate the impact of the various processes of social mobility on health inequalities, mortality was observed over the period 1991-1999. The study population consists of men and women aged 25-49 at the beginning of mortality follow-up (1991), and registered as resident in Turin at both the 1981 and the 1991 censuses (N = 127,384). Health status was determined by observing hospital admission. For the purpose of the study healthy individuals were those with no hospital admissions during the period 1984-1986, while those admitted were classed as unhealthy. Social mobility in the labour market was measured via an interval data index of upward and downward movements on a scale of social desirability of occupations, designed for the Italian labour force via an empirical study carried out by de Lillo and Schizzerotto (La valutazione sociale delle occupazioni. Una scala di stratificazione occupazionale per l'Italia contemporanea, Il Mulino, Bologna, 1985). Movement out of the labour market was described by a discrete variable with four conditions: employed, unemployed, early retired and women returning from work to the housewife status. The relationship between health status and occupational mobility was analysed via analysis of variance and multinomial logistic regression. Health inequalities were measured by the ratio of standardised mortality rates in the unskilled working class and the upper middle class. The study found a weak relationship between health status and occupational mobility chances. Decidedly stronger was the impact on occupational mobility of gender, education and "ethnicity" (being born in the South of Italy). The relationship between occupational mobility and health takes two different forms. Occupational mobility in the labour market decreases health inequalities; occupational mobility out of the labour market (early retirement, unemployment, housewife return) widens them. The maximum contribution health-related intra generational social mobility can make towards health inequalities was estimated at about 13% for men. PMID- 14759701 TI - Abstracts of the XIIth Biennial Winter Workshop on Schizophrenia. Davos, Switzerland, February 7-13, 2004. PMID- 14759702 TI - Molecular cloning, nucleotide sequencing and expression of genes encoding a cytochrome P450 system involved in secondary amine utilization in Mycobacterium sp. strain RP1. AB - Mycobacterium sp. strain RP1 degrades morpholine, piperidine, and pyrrolidine and is able to use these compounds as the sole source of carbon, nitrogen, and energy. Cytochrome P450 (MorA) is involved in the biodegradation of these secondary amines. A 3.9-PstI genomic DNA fragment, containing the gene encoding MorA, was cloned and sequenced. Four open reading frames were detected on this DNA fragment. The first encoded a cytochrome P450 designated as MorA which was the second member of the CYP151 family and was named CYP151A2. The second open reading frame (morB) featured a [3Fe-4S] type of ferredoxin. A third gene (morC), exhibiting sequence identity to known reductases, and a fourth truncated gene encoding a putative glutamine reductase (orf1' ), were found downstream of morB. Recombinant MorA cytochrome P450 was purified to homogeneity from Escherichia coli. The purified enzyme was a monomeric soluble protein with an apparent Mr of about 45,000. CYP151A2 catalyzed the ring cleavage of the secondary amines and the Vmax/KMapp values indicated that pyrrolidine is the preferred substrate for this monooxygenase. PMID- 14759703 TI - Quantification of human astroviruses in sewage using real-time RT-PCR. AB - Human astroviruses constitute a significant cause of acute diarrhea in children. Viral transmission occurs via the fecal-oral route, predominantly person-to person, but the consumption of fecally contaminated water and shellfish has also been implicated. Viral pathogen detection in water, especially, in wastewater, is difficult and PCR is widely used to detect these viruses. Despite the recent development of real-time quantitative PCR, quantification of astroviruses in sewage had not been available up to now. We have developed a method to quantify astroviruses in sewage. For this purpose, we designed a set of primers and a fluorogenic probe located at the 3' -end of the genome of human astroviruses. The amplified region was cloned and the plasmid was transcribed to generate calibration standards for quantification. After validation of the standards, the method was evaluated in artificially contaminated samples. To validate the method on naturally contaminated samples, raw and treated wastewater samples were collected monthly for one year in a sewage treatment plant. Astrovirus genomes were detected in all samples collected at the entrance to the sewage treatment plant, with a mean value of 4.1 x 10(6) astrovirus genomes for 100 ml. Effluents were less strongly contaminated, with a mean value of 1.01 x 10(4) astrovirus genomes. The high prevalence of astroviruses in sewage treatment plant effluents indicates that these plants are not efficiently eliminating the virus. This is a major public health concern and new techniques of depuration are needed. Our method could be effectively used in evaluating new treatment processes to reduce the viral load in the effluent of treatment plants. PMID- 14759704 TI - Discrimination of Klebsiella pneumoniae and Klebsiella oxytoca phylogenetic groups and other Klebsiella species by use of amplified fragment length polymorphism. AB - Bacteria of the genus Klebsiella are opportunistic pathogens responsible for an increasing number of multiresistant infections in hospitals. The two clinically and epidemiologically most important species, Klebsiella pneumoniae and K. oxytoca, have recently been shown to be subdivided into three and two phylogenetic groups, respectively. The aim of this study was an in depth evaluation of the amplified fragment length polymorphism (AFLP) genetic characterization method for epidemiological and phylogenic analyzes of Klebsiella isolates. First, we investigated the variability of AFLP patterns for Klebsiella strains within and between different outbreaks. Second, by use of carefully characterized phylogenetically representative strains, we examined whether different Klebsiella species and phylogenetic groups can be discriminated using AFLP. Twenty-four strains originating from seven presumed outbreaks and 31 non associated strains were investigated. The AFLP fingerprints of all epidemiologically associated strains showed three or fewer fragment differences, whereas unrelated strains differed by at least four fragments. Cluster analysis of the AFLP data revealed a very high concordance with the phylogenetic assignation of strains based on the gyrA sequence and ribotyping data. The species K. pneumoniae, K. oxytoca, K. terrigena and the possibly synonymous pair K. planticola/K. ornithinolytica each formed a separate cluster. Similarly, strains of the phylogenetic groups of K. pneumoniae and K. oxytoca fell into their corresponding clusters, with only two exceptions. This study provides a preliminary cut-off value for distinguishing epidemiologically non-related Klebsiella isolates based on AFLP data; it confirms the sharp delineation of the recently identified phylogenetic groups, and demonstrates that AFLP is suitable for identification of Klebsiella species and phylogenetic groups. PMID- 14759705 TI - Morphological and intracellular alterations induced by Serratia marcescens cytotoxin. AB - In the present work, in vitro assays were used to investigate the toxicity of Serratia marcescens cytotoxin in cultured Chinese hamster ovary (CHO) cells. The time necessary to detect cellular alterations such as the onset of apoptosis, the perturbation of mitochondrial function, and cytoskeletal changes was assessed. The internalization of the cytotoxin by CHO cells was also examined. Within 10-15 min of exposure to cytotoxin, CHO cells became round, the nucleus shrank, the chromatin became more compact, and cytoplasmic blebs appeared on the cell surface. TUNEL (TdT-mediated dUTP nick end labeling) and propidium iodide staining identified some nuclei with fragmented DNA, and electrophoresis of CHO cell DNA obtained after 30-min exposure to S. marcescens toxin showed a pattern of DNA fragments typically associated with apoptosis. The cells also lost their characteristic actin organization within 10 min of exposure to cytotoxin. Lactate dehydrogenase leakage was detected after 20-min exposure to the cytotoxin and increased with time thereafter. Concomitantly, there was a time-dependent reduction in mitochondrial activity. Fluorescein-labeled S. marcescens cytotoxin was detected only on the surface of CHO cells, even after 30-min exposure to the toxin. These results show that there was no internalization of the toxin by CHO cells, and that, once bound to the cell surface, the toxin was able to induce changes in intracellular metabolism and to trigger cell death by apoptosis. PMID- 14759706 TI - In vitro activation of the hemolysin in Prevotella nigrescens ATCC 33563 and Prevotella intermedia ATCC 25611. AB - Hemolytic activity was evaluated in the putative periodontopathogens Prevotella intermedia and Prevotella nigrescens. Whole cells of both species present weak hemolytic activity evidenced only by solid media assays after 48 h of bacterial growth or after 5 h of interaction with erythrocytes at 37 degrees C in liquid assays. In this work we show that the use of crude extract allowed the detection of a higher hemolytic activity for P. intermedia, but surprisingly not for P. nigrescens. Incubation at 37 degrees C for 9 h, or treatment with trypsin or proteinase K, increased or exposed the hemolytic activity of P. intermedia and P. nigrescens crude extract, respectively. The activation process was inhibited by TLCK and PMSF but not by EDTA, E-64 or pepstatin A, indicating the serino protease nature of the factor involved in activation of P. intermedia and P. nigrescens hemolysins. Both the buffer and the pH employed for cell fractionation influenced the activation of hemolysin, and the best results were obtained with Universal buffer at pH 8.0. The activated hemolysins acted optimally at pH 6.5 at 37 degrees C and the maximum hemolytic activity was detected at the early log phase of growth. The results of this study show for the first time a strong hemolytic activity for P. nigrescens and evidence of proteolytic activation of hemolysins produced by periodontopathogens. PMID- 14759707 TI - Regulation of the cytotoxic effects of Pseudomonas fluorescens by growth temperature. AB - We had previously shown that the psychrotrophic bacterium Pseudomonas fluorescens can act as a pathogen, inducing apoptosis and necrosis in neurons and glial cells. In the present study, we investigated the influence of the growth temperature of P. fluorescens on its infectious potential. Adherence of P. fluorescens to glial cells was found to be maximal with bacteria grown at a low temperature (8 degrees C). At that temperature the swimming behaviour was markedly reduced. An increase in the growth temperature to 19, 28 or 32 degrees C strongly diminished the binding of bacteria to host cells. Thus, the adhesion phenotype of P. fluorescens appears to be independent of the motility of the bacteria. The apoptotic effect of P. fluorescens, determined by morphological (nuclear condensation) and biochemical (induction of nitric oxide synthase activity) indicators, correlated well with its binding activity on glial cells. In contrast, there was a clear dissociation between maximum binding and maximal necrotic action (measured by the release of lactate dehydrogenase) observed with bacteria grown at 19 degrees C. As suggested by capillary electrophoresis analysis, the differences in apoptotic effects may be related to variations in the molecular structure of LPS originating from bacteria grown at low and high temperatures, whereas the necrotic effect, which was maximal at the optimum temperature for the secretion of exoenzymes, could reflect variations in the metabolic activity of bacteria. PMID- 14759708 TI - A novel alpha-Proteobacterium, Nordella oligomobilis gen. nov., sp. nov., isolated by using amoebal co-cultures. AB - Using an amoebal co-culture procedure, a novel alpha-Proteobacterium phylogenetically close to two uncultured aquatic bacteria was isolated. On the basis of phenotypic characterization and 16S rRNA gene sequence analysis, we propose the new genus and species Nordella oligomobilis gen. nov., sp. nov. The genus Nordella forms a well separated taxon in the order Rhizobiales within the alpha-2 subgroup of Proteobacteria. Its close relatives are environmental uncultured bacteria. PMID- 14759709 TI - Possible role of nitric oxide in the biology of breast carcinoma: review of the available literature. AB - Nitric oxide was studied to investigate its possible involvement in the promotion of breast carcinoma: both the development of the primary tumour and the process of metastasis seem to be influenced by the presence and the amount of nitric oxide. We review the available literature on this topic, which seems to suggest an influence of nitric oxide on the cancer cell biology in breast carcinoma, but the argument is still controversial. More studies are needed to clarify the sequence of events and the real impact of nitric oxide on the behaviour of the disease. PMID- 14759710 TI - Changes in survival after breast cancer: improvements in diagnosis or treatment? AB - We have compared 5-year survival rates in two cohorts of women diagnosed with breast cancer in Brisbane, Australia, between 1981-1984 and 1990-1994. Tumours diagnosed in the early 1990s were significantly smaller and less likely to have nodal involvement than those diagnosed 10 years earlier (P<0.0001). The size difference was particularly striking for women aged over 50 at diagnosis, those targeted for screening. Five-year survival was greater among women diagnosed in the 1990s (84% vs. 74%; hazard ratio (HR) 0.61, 95% confidence interval (CI) 0.46 0.81). After adjusting for the effects of tumour size and nodal status this difference was reduced, but women diagnosed more recently still showed improved survival (HR 0.75; 95% CI 0.56-1.01) and disease-free survival (HR 0.72; 0.56 0.92) at 5 years. This suggests that both earlier diagnosis and changes in breast cancer treatment have contributed to improved breast cancer survival. PMID- 14759711 TI - Detection by screening mammography is a powerful independent predictor of survival in women diagnosed with breast cancer. AB - Four hundred and sixteen invasive breast cancers, detected initially by mammography, were compared with 929 presenting symptomatically, all treated at a South Australian teaching hospital. Predictable differences included lower stages and grades, less vascular invasion and proliferative activity, and more hormone receptor expression among the mammographically detected. Unpredicted differences included significantly higher survivals for mammographically detected cases throughout the 9 year follow-up period after adjusting for stage and the Nottingham Prognostic Index. In a multivariable analysis, differences in stage, grade, and hormone receptor expression accounted for only about half the survival advantage of mammographically detected tumours. Accounting for additional person and tumour characteristics had only a marginal effect on this result. This suggests that detection by mammography has independent favourable prognostic significance beyond that explained by conventional indicators. If confirmed, this finding would have important implications for the prognostic advice given to women and may merit further investigation into its underlying biological mechanisms. PMID- 14759712 TI - Prognostic indexes in breast cancer: comparison of the Nottingham and Adelaide indexes. AB - The search for single independent prognostic factors in breast cancer has often produced conflicting results. Therefore, prognostic indexes have been compiled by combining several parameters. In this study we compare the Nottingham Prognostic Index (NPI), which is based on traditional prognostic factors (diameter of the neoplasm, lymph node status and histological grade) with the Adelaide Prognostic Index (API), which is based on the tumour diameter and two biological parameters: oestrogen receptors and cell kinetics. We considered 82 cases of breast cancer observed over the period 1987-1990 with a minimum follow-up of 60 months. The NPI gives a better definition of the prognostic profile for each patient. Our results indicate three prognostic groups (good, moderate, unfavourable), which differ with respect to disease-free survival (DFS; P=0.0024) and overall survival (OS; P=0.0033). In contrast, the API scores showed no significant correlation with OS or DFS. The use of prognostic indexes, especially when compiled using traditional parameters, is a useful aid to the clinician, since they can provide a reliable indication of how individual tumours will evolve. PMID- 14759713 TI - CYP2C and IL-6 expression in breast cancer. AB - Besides hepatic P450 (cytochrome P450) metabolism, there is increasing interest in the possibility of intratumoral activation of oxazaphosphorines by P450. Therefore, we investigated the expression of P450 (CYP2C8, CYP2C9, CYP2C18, and CYP2C19) by RT (reverse transcriptase)-polymerase chain reaction (PCR) and of CYP2C9 by Western blotting in 10 different breast tumor samples. Since P450 may be down regulated by interleukin (IL) IL-6, the receptor (R) for IL-6 was analyzed by RT-PCR and IL-6 in supernatants was calculated from ELISA data. None of the breast tumors was positive for CYP2C18 and CYP2C19 mRNA, whereas CYP2C8 and CYP2C9 were detected in all 10 breast tumors. Correspondingly, all breast tumors tested (9 of 10) revealed low, but nevertheless positive, staining of the CYP2C9 protein. All 10 samples were positive for the IL-6 receptor mRNA. ELISA measurement of IL-6 cytokine in supernatants revealed that all measured samples (8 of 10) were producing IL-6, the amounts ranging from 0.004 to 3.1 ng/g(tumor tissue). In summary, we have demonstrated that tumors of the breast express two out of four members of the CYP2C family, indicating that activation of such prodrugs as oxazaphosphorines may take place intratumorally. The presence of the IL-6 receptor and of IL-6 cytokine, which is produced in an autocrine manner, opens up the possibility that the well-known down regulating effect of IL-6 also takes place in breast tumors and might explain the weak or even absent expression of different CYP2C members. PMID- 14759714 TI - Identification of carcinoma cells in peripheral blood samples of patients with advanced breast carcinoma using RT-PCR amplification of CK7 and MUC1. AB - We have undertaken a pilot study to attempt to identify circulating carcinoma cells in a series of patients with advanced breast carcinoma, using reverse transcription-polymerase chain reaction (RT-PCR) to amplify mRNA of epithelial specific antigens. Using this method to amplify mRNA of MUC1 and cytokeratin 7 (CK7) the sensitivity of the technique was demonstrated by means of diluted concentrations of "spiked MCF7" cells in whole blood, showing a detection limit of 1 in 10(6) (CK7) and 1 in 10(5) (MUC1). Positive results were obtained from the peripheral blood of all nine female patients with advanced breast cancer for CK7 and eight of the nine patients for MUC1. CK7 was however detected in five of 11 healthy controls (eight females, three males) and MUC1 in one of the 11 controls. None of the control group were positive for both CK7 and MUC1, in contrast to eight of the nine patients with advanced breast carcinoma who were positive for both markers. The RT-PCR method thus appears sufficiently sensitive to identify circulating tumour cells in peripheral blood samples from patients with advanced breast carcinoma. However a high proportion of false-positive results was seen in the control population. More extensive investigation is required before the technique is likely to be of benefit clinically. PMID- 14759715 TI - False-negative frozen section of sentinel lymph node biopsy for breast cancer. AB - A prospective study is presented of frozen section examinations (FS) performed in parallel with 265 consecutive sentinel lymph node procedures (SLNP) over a 20 month period. The final pathological study included immunohistochemistry (IHC) for keratin if the haematoxylin-eosin (HE)-stained section was tumour free. FS correctly identified node-positive or node-negative axillae in 235 cases. In 28 SLNPs the final examination gave a positive result not detected in the FS, resulting in reoperation. In 21 of these false-negative (FN) cases micrometastases (MIM) were present. There were no false-positive cases, but in two cases of lobular carcinoma the findings in the FS were equivocal, the final reports recording metastases in one but not in the other. Lobular carcinoma and other less common subtypes of carcinoma were overrepresented, ductal carcinoma not otherwise specified (NOS) being less likely to affect the FN findings. PMID- 14759716 TI - Detection of cancer cells in the axillary drainage using RT-PCR after operations for breast cancer. AB - The object of this study was to examine whether MUC-1 can be detected in the axillary lymphatic drainage of patients who have undergone conservative surgery for breast cancer and to assess the correlations between the presence of MUC-1 and prognostic factors in breast cancer. Sixty-eight women with invasive ductal carcinoma of the breast underwent wide local excision and axillary lymph node dissection. Axillary drains were inserted in all these cases, and the presence of MUC-1 and beta-actin was evaluated by RT-PCR in the lymphatic fluid collected after the operation. Prognostic factors included tumour size and grade, vascular and lymphatic invasion, clearance margins of the resected specimens and status of the axillary lymph nodes. RT-PCR assays for MUC-1 in the axillary fluid were positive in 17 patients (25%). The presence of MUC-1 was associated with increased tumour size and showed a positive correlation with axillary lymph node metastases and incomplete resection of the tumour. RT-PCR can disclose cancer cells in the axillary fluid after conservative surgery for breast cancer. The presence of MUC-1 in the axillary drainage may be associated with poor prognostic features, and its detection may have implications for therapy as it suggests that re-excision should be considered. PMID- 14759717 TI - Prospective estimation of rates of change in mammographic parenchymal patterns: influence of age and of hormone replacement therapy. AB - The objective of this study was to assess the effect of age, breast size and use of hormone replacement therapy (HRT) on the rate of change of mammographic parenchymal patterns, and the effect of age on the probability of misclassification between patterns. It was designed as a longitudinal study of the members of the treatment arm of a non-randomized screening trial in which subjects were assigned to screening or not by year of birth. The subjects were women in the Kotka district of Finland, each of whom attended for four or five mammographic screens. Participants were all women living in the district who were born in the relevant years and accepted an invitation to screening. A model was fitted to the longitudinal data comprising the observed Wolfe patterns on each woman, with age and breast size as predictors of breast density at first screen, age and HRT use as predictors of change in density at future screens, and age as a predictor of misclassification of true density between favourable (non-dense) and unfavourable (dense) patterns (according to the Wolfe classification). Relevant posterior probability estimates (with 95% credible intervals) were as follows. The probability that a woman of age 43.5 is truly in the favourable state ranged from 0.35 (0.34-0.37) for smallest breast size to 0.74 (0.72-0.76) for the largest. The probability that a woman is truly in the favourable state at first screen increased from 0.37 (0.36-0.38) at age 40 to 0.59 (0.58-0.60) at age 47. The probability that a woman having a later screen who had truly been in the unfavourable state at her previous screen changed to the favourable state increased from 0.12 (0.11-0.13) at age 42 to 0.48 (0.46-0.50) at age 55 for a woman not taking HRT, and from 0.10 (0.09-0.11) to 0.43 (0.40-0.45) at the same ages for a woman taking HRT. The probability that a woman would have changed from being truly in the favourable state to the unfavourable state was 0.003 (0.001 0.003) for any age and HRT use. The probability that a woman truly in a favourable state would be correctly classified rose from 0.87 (0.85-0.89) at age 40 to 0.998 (0.997-0.998) at age 55. The probability that a woman truly in the unfavourable state would be correctly classified decreased from 0.96 (0.95-0.97) to 0.93 (0.91-0.94) between the same ages. The probability of being in a non dense, favourable state increases with age, as does the rate of change from dense to non-dense patterns. These are consistent with previous work. The probability of non-dense patterns and the rate of change to non-dense patterns are reduced with HRT use. Errors of classification are relatively rare, but are dependent on the age of the subject. PMID- 14759718 TI - Management of physiological gynaecomastia with tamoxifen. AB - AIMS: We aimed to confirm suggestions that tamoxifen therapy alone may resolve physiological gynaecomastia. METHODS: A prospective audit of the outcome of tamoxifen routinely given to men with physiological gynaecomastia was carried out at Nottingham. Men referred with gynaecomastia had clinical signs recorded, e.g., type (diffuse 'fatty' or retro-areolar 'lump'), size and possible aetiology. They were offered oral tamoxifen 20mg once daily for 6-12 weeks. On follow-up patients were assessed for complete resolution (CR), partial resolution where patient is satisfied with outcome (PR) or no resolution (NR). Success was either CR or PR. RESULTS: Thirty-six men accepted tamoxifen for physiological gynaecomastia. Median age was 31 (range 18-64). Tenderness was present in 25 (71%) cases. Sixteen men (45%) had 'fatty' gynaecomastia and 20 had 'lump' gynaecomastia. Tamoxifen resolved the mass in 30 patients (83.3%; CR=22, PR=8) and tenderness in 21 cases (84%; CR=0, PR=0). Lump gynaecomastia was more responsive to tamoxifen than the fatty type (100% vs. 62.5%; P=0.0041). CONCLUSIONS: Oral tamoxifen is an effective treatment for physiological gynaecomastia, especially for the lump type. PMID- 14759720 TI - Breast cancer presenting initially with urinary incontinence: a case of bladder metastasis from breast cancer. AB - Bladder metastasis from breast cancer is rare. Patients with breast cancer, in particular patients with a lobular carcinoma subtype, who present with urinary symptoms including incontinence, hematuria, dysuria, and frequency should have the possibility of bladder metastases kept in mind and investigated with cystoscopy and imaging as necessary. PMID- 14759719 TI - Metastatic breast cancer response after Exemestane withdrawal: a case report. AB - A forty-three year old female patient with right sided breast cancer was treated with adjuvant radiotherapy and Tamoxifen. She had a relapse in the right supraclavicular fossa after a disease free period of 12 years. She progressed, after a short period of disease stabilization on Idoxifene followed by a further stable period on Anastrazole. At further disease relapse, she was commenced on Exemestane. After a period of disease stabilization and subsequent disease progression in the right supraclavicular fossa, Exemestane was discontinued. Six weeks later there was a marked regression of the right supraclavicular mass. Withdrawal response to Tamoxifen is well documented. To our knowledge a similar response after Exemestane withdrawal has not been reported in the literature. PMID- 14759721 TI - Primitive leiomyosarcoma of the breast: case report and review of the literature. AB - Sarcomas of the breast account for under 1% of breast tumours. Leiomyosarcomas are less common, being a subgroup of sarcomas of the breast. Only 23 cases with immunohistochemical or electron microscopy confirmation are reported in the literature. The case of a 58-year-old woman with a leiomyosarcoma 4 cm in diameter in the upper external quadrant of the right breast is presented. Mammography and sonography were compatible with a fibroadenoma or phylloides tumour, and fine-needle aspiration (FNA) suggested medullary carcinoma. The patient underwent a Madden-type modified radical mastectomy and axillary lymphadenectomy. Adjuvant chemotherapy was implemented with Adriamycin (four cycles of 21 days). One year after surgery the patient is tumour free. PMID- 14759722 TI - Gynaecomastia, neurofibromatosis and breast cancer. AB - Gynaecomastia commonly affects pubertal boys. We report a rare case of bilateral ductal carcinoma in situ found incidentally after breast reduction surgery in a young man with neurofibromatosis. PMID- 14759723 TI - PTTG mRNA expression in primary breast cancer: a prognostic marker for lymph node invasion and tumor recurrence. AB - The proto-oncogene pituitary tumor-transforming gene (PTTG) is described as abundantly overexpressed in a variety of neoplasms and cancer cell lines. We analyzed PTTG mRNA expression in 72 tumor samples derived from primary tumors of patients suffering from breast cancer and in unaffected breast epithelium, checking for correlations between expression levels and standard clinico pathological parameters over a 5-year observation period. We found a direct correlation between PTTG mRNA overexpression and lymph node infiltration. Furthermore, overexpression of this gene in tumors correlated with a higher degree of tumor recurrence within the subsequent observation period. Taken together, these results demonstrate that determination of PTTG expression in primary tumors of the breast is a powerful tool for the assessment of potential tumor aggressiveness. PMID- 14759724 TI - The future of breast surgery: a new sub-speciality of oncoplastic breast surgeons? PMID- 14759725 TI - Is it time for a paradigm shift in the training of breast surgeons as a sub speciality? PMID- 14759726 TI - Studies on some glitazones having pyridine as the linker unit. AB - Molecular modeling on various well-known glitazones carrying a pyridine ring instead of benzene ring as the middle linker unit showed conformational rigidity as compared to their parent molecules. Blocking the lone pair of electrons on the pyridine N, made them flexible once again. A few representatives of these analogues were synthesized and their efficacy as PPARgamma agonists evaluated. PMID- 14759727 TI - Synthesis, biological activity, QSAR and QSPR study of 2-aminobenzimidazole derivatives as potent H3-antagonists. AB - We report the design, synthesis, QSPR and QSAR of a new class of H(3) antagonists, having a 2-aminobenzimidazole moiety connected to the 4(5) position of an imidazole ring through di- or tri-methylene chains. Eleven substituents, selected by experimental design to obtain broad and non-correlated variation in their lipophilic, electronic and steric properties, were introduced at the 5(6) position of the benzimidazole nucleus. The compounds were tested for their H(3) receptor affinity, by displacement of [(3)H]-(R)-alpha-methylhistamine ([(3)H] RAMHA) binding to rat brain membranes (pK(i)), for intrinsic activity, evaluating their effect on [(35)S]GTPgammaS binding to rat brain membranes, and for H(3) antagonist potency, on electrically stimulated guinea-pig ileum (pK(B)). The pK(i) values of the derivatives with longer chain (5a-k) ranged over 2 orders of magnitude, with the 5(6)-methoxy derivative 5d endowed with sub-nanomolar affinity (pK(i)=9.37). The series having two methylene groups in the chain spacer (4a-k), showing a small variation in affinity, revealed to be somewhat insensitive to ring substitution. Lipophilicity (log P) and basicity (pK(a)) of the newly synthesized compounds were measured and related to receptor affinity in a QSAR study. Multiple regression analysis (MRA) showed an approximate parabolic dependence of pK(i) on log P, while an additional electronic effect of the substituents on benzimidazole tautomerism is suspected. PMID- 14759728 TI - A new linker for glucuronylated anticancer prodrugs. AB - The synthesis, enzymatic hydrolysis and self decomposition of model glucuronylated prodrugs, incorporating a new linker with different aryl substituents, have been studied. Determination of kinetic parameters (V(max), K(m) and t(1/2)) showed the important role of aromatic substitution in enzymatic recognition and linker decomposition. PMID- 14759729 TI - Analogues of thiolactomycin as potential anti-malarial and anti-trypanosomal agents. AB - A series of analogues of the naturally occurring antibiotic thiolactomycin (TLM) have been synthesised and evaluated for their ability to inhibit the growth of the malaria parasite, Plasmodium falciparum. Thiolactomycin is an inhibitor of Type II fatty acid synthase which is found in plants and most prokaryotes, but not an inhibitor of Type I fatty acid synthase in mammals. A number of the analogues showed inhibition equal to or greater than TLM. The introduction of hydrophobic alkyl groups at the C3 and C5 positions of the thiolactone ring lead to increased inhibition, the best showing a fourteenfold increase in activity over TLM. In addition, some of the analogues showed activity when assayed against the parasitic protozoa, Trypanosoma cruzi and Trypanosoma brucei. PMID- 14759730 TI - Construction of a chemically and conformationally self-replicating system of amyloid-like fibrils. AB - The amyloid-like fibril is considered to be a macromolecular self-assemblage with a highly-ordered quaternary structure, in which numerous beta-stranded polypeptide chains align regularly. Therefore, this kind of fibril has the potential to be engineered into proteinaceous materials, although conformational alteration of proteins from their native form to the amyloid form is a misfolding and undesirable process related to amyloid diseases. In this study, we have attempted to design an artificial system to explore applicability of using the amyloid-like fibril as a construct possessing self-recognition and self-catalytic abilities. A peptide self-replicating system based on the beta-structure of the amyloid-like fibril was designed and constructed. The beta-stranded peptide was self-replicated by the native chemical ligation reaction, and the newly generated peptide was self-assembled into amyloid-like fibrils. Thus, the constructed system was of both chemical and conformational self-replicating fibrils. PMID- 14759731 TI - QSAR and kinetics of the inhibition of benzaldehyde derivatives against Sacrophaga neobelliaria phenoloxidase. AB - A group of 18 benzaldehyde derivatives was characterized as a family of mixed type inhibitors on the oxidation of l-3, 4-dihydroxyphenylalanine (l-DOPA) catalyzed by Sacrophaga neobelliaria phenoloxidase (PO), presumably by forming a Schiff base with a primary amino group of the enzyme. Inhibition constants and IC(50) were determined. Cuminaldehyde was the most active inhibitor with an IC(50) of 0.0067 mM. Vanillin was the least active inhibitor with an IC(50) of 38 mM. Four physicochemical descriptors were identified by stepwise multiple regressions as significant predictors on the inhibition activity, which were further rotated by principle component analysis yielding two significant principle properties. It was shown that hydrophobicity of the substituent at the para position of the aldehyde group played major role on inhibition activity: one unit increase in Hansch-Fujita pi value of the substituent led to about 4.5 [95% confidence interval is (7.9, 2.6)] fold increase on IC(50). Electron-donating effect of the substituent at the para position of the aldehyde group was less important than hydrophobicity. Hydroxyl group at the ortho position of the aldehyde group contributed to higher inhibition activity, presumably by forming a quasi-six-membered ring with the unshared pair of electrons on the nitrogen atom of the amino group through intramolecular hydrogen bonding. PMID- 14759732 TI - Synthesis and SAR exploration of dinapsoline analogues. AB - Dinapsoline is a full D(1) dopamine receptor agonist that produces robust rotational activity in the unilateral 6-OHDA rat model. This compound is orally active, and shows a low tendency to cause tolerance in rat models. The active enantiomer was determined to have the S-(+) configuration, and the opposite enantiomer is essentially devoid of biological activity. Taken together, dinapsoline has significant metabolic and pharmacological advantages over previous D(1) agonists. In an attempt to define the structure-activity relationships (SARs) and to map out the key elements surrounding the unique structure of dinapsoline, core analogues and substitution analogues of the parent tetracyclic condensed ring structure were prepared. Based on a recently developed synthesis of dinapsoline and its enantiomers, both core and substitution analogues on all four rings (A, B', C and D ring) of dinapsoline were synthesized. It was found that affinity for both D(1)and D(2) receptors was decreased by most substituents on the A, B', and C rings, whereas D ring substitutions preserved much of the dopamine receptor binding activity. PMID- 14759733 TI - A novel approach to predict a toxicological property of aromatic compounds in the Tetrahymena pyriformis. AB - The TOPological Substructural MOlecular DEsign (TOPS-MODE) has been successfully used in order to explain the toxicity in the Tetrahymena pyriformis on a large data set. The obtained models for the training set had good statistical parameters (R(2)=0.72-0.81, p<0.05) an also the prediction power of the models found was adequate (Q(2)=0.70-0.80). A detailed study of the influence of variable numbers in the equation and the statistical outliers was carried out; leading to a good final model with a better physicochemical interpretation than the rest of the published models. Only two molecular descriptors codifying dipolar and hydrophobic features were introduced. Finally, the fragment contributions to the toxicity prediction evidenced the powerful of this topological approach. PMID- 14759734 TI - QSAR modeling of HIV-1 reverse transcriptase inhibitor 2-amino-6 arylsulfonylbenzonitriles and congeners using molecular connectivity and E-state parameters. AB - Anti-HIV-1 activity (assayed in MT-4 cell line) and HIV-1 reverse transcriptase (RT) binding affinity of 2-amino-6-arylsulfonylbenzonitriles and their thio and sulfinyl congeners (Chan et al., J. Med. Chem., 2001, 44, 1866) have been modeled using E-state index along with molecular connectivity and indicator parameters in an attempt to explore the different fragments of the molecules contributing significantly to the activities. Stepwise multiple regression procedure was adopted to develop the topological models. The models generated were of acceptable statistical quality and predictive potential. The results show that for both the response variables, first order fragmental valence molecular connectivity of the meta substituents of the aryl ring plays a significant role: second meta substituents show supraadditive action on the activities probably due to enhanced binding (presumably through dispersion interaction) of the ligand with the binding site. Again, presence of sulfone moiety contributes significantly to the activities. Further, presence of meta-trifluoromethyl group at the aryl ring is detrimental for both the activity parameters. Additionally, the anti-HIV-1 model shows specific contributions of the E-state values of different atoms and positive contribution of the ortho-methoxy group present on the aryl ring. PMID- 14759735 TI - Antimalarial activity of N(6)-substituted adenosine derivatives. Part 3. AB - A series of novel 3'-amido-3'-deoxy-N(6)-(1-naphthylmethyl)adenosines was synthesized applying a polymer-assisted solution phase (PASP) protocol and was tested for anti-malarial activity versus the Dd2 strain of Plasmodium falciparum. Further, this series and 62 adenosine derivatives were analyzed regarding 1-deoxy d-xylulose 5-phosphate (DOXP) reductoisomerase inhibition. Biological evaluations revealed that the investigated 3',N(6)-disubstituted adenosine derivatives displayed moderate but significant activity against the P. falciparum parasite in the low-micromolar range. On the molecular level, DOXP reductoisomerase utilizing an adenosyl-containing substrate was identified as a promising metabolic target for ligands of adenosine binding motifs. PMID- 14759736 TI - Synthesis and biological activity of isopentenyl diphosphate analogues. AB - A series of analogues of isopentenyl diphosphate (IPP) having a dicarboxylate moiety in place of the diphosphate were synthesized and investigated as inhibitors of undecaprenyl diphosphate (UPP) synthase and protein farnesyltransferase (PFTase). PFTase is involved in control of cell proliferation and is known to be inhibited by certain maleic acid derivatives bearing long alkyl substituents (> or =12 carbons, e.g., chaetomellic acid). UPP synthase is a potential target for antimicrobial agents and utilizes isopentenyl diphosphate (IPP) as a substrate. A number of dicarboxylate-containing IPP analogues were prepared in 2-5 steps from commercially available starting materials with good overall yield (20-78%). These syntheses involved the conjugate addition of an organocuprate to dimethyl acetylenedicarboxylate (DMAD) followed by basic ester hydrolysis. The E-pentenylbutanedioic acid 32 showed inhibition of UPP synthase with an IC(50) of 135 microM. Compound 30 displays competitive inhibition of PFTase with a K(i) of 287 microM. PMID- 14759737 TI - Synthesis and antiproliferative activity of some new DNA-targeted alkylating pyrroloquinolines. AB - Two novel DNA-direct alkylating agents, consisting of aniline mustard linked to an angular 3H-pyrrolo[3,2-f]quinoline nucleus, were synthetized and assayed for their in vitro antiproliferative activity. Simple convergent synthesis, consisting of separate preparation of 9-chloro-3H-pyrrolo[3,2-f]quinoline and p amino-aniline derivatives, and following their linkage by substitution reactions 8a, b and 10, yielded the corresponding diol derivatives 7b and 9. Biological properties were evaluated with respect to cell growth inhibition, ability to form cross-links with DNA, and capacity to give rise to a molecular complex with the macromolecule for 7b, 8b, 9 and 10. PMID- 14759738 TI - Design of tetrapeptide ligands as inhibitors of the Src SH2 domain. AB - Src homology-2 (SH2) domains are noncatalytic motifs containing approximately 100 amino acid residues that are involved in intracellular signal transduction. The phosphotyrosine-containing tetrapeptide pTyr-Glu-Glu-Ile (pYEEI) binds to Src SH2 domain with high affinity (K(d)=100 nM). The development of five classes of tetrapeptides as inhibitors for the Src SH2 domain is described. Peptides were prepared via solid-phase peptide synthesis and tested for affinity to Src SH2 domain using a fluorescence polarization based assay. All of the N-terminal substituted pYEEI derivatives (class II) presented binding affinity (IC(50)=of 2.7-8.6 microM) comparable to pYEEI (IC(50)=6.5 microM) in this assay. C-Terminal substituted pYEEI derivatives (class III) showed a lower binding affinity with IC(50) values of 34-41 microM. Amino-substituted phenylalanine derivatives (class IV) showed weak binding affinities (IC(50)=16-153 microM). Other substitutions on phenyl ring (class I) or the replacement of the phenyl ring with other cyclic groups (class V) dramatically decreased the binding of tetrapeptides to Src SH2 (IC(50)>100 microM). The ability of pYEEI and several of the tetrapeptides to inhibit the growth of cancer cells were assessed in a cell-based proliferation assay in human embryonic kidney (HEK) 293 tumor cells. The binding affinity of several of tested compounds against Src SH2 domain correlates with antiproliferative activity in 293T cells. None of the compounds showed any significant antifungal activity against Candida albicans ATCC 14053 at the maximum tested concentration of 10 microM. Overall, these results provided the structure-activity relationships for some FEEI and YEEI derivatives designed as Src SH2 domain inhibitors. PMID- 14759739 TI - QSAR study on benzenesulphonamide carbonic anhydrase inhibitors: topological approach using Balaban index. AB - QSAR study on benzenesulphonamide carbonic anhydrase inhibitors has been made using the most discriminatory Balaban index (J). The regression analysis has shown that even in monoparametric regression this index gave excellent results. Furthermore, using the combination of the Balaban Index (J) with the first-order Randic connectivity index ((1)chi) and indicator parameters, tremendous improvement in the statistics has been observed. The results are critically discussed on the basis of regression data and cross-validation parameters. PMID- 14759740 TI - 11H-Isoquino[4,3-c]cinnolin-12-ones; novel anticancer agents with potent topoisomerase I-targeting activity and cytotoxicity. AB - Recent studies have identified 2,3-dimethoxy-8,9-methylenedioxy-11-[(2 dimethylamino)ethyl]-11H-isoquino[4,3-c]cinnolin-12-one (1a) as a novel topoisomerase I-targeting agent with potent cytotoxic activity. The effect of varied substituents at the 11-position of 2,3-dimethoxy-8,9-methylenedioxy-11H isoquino[4,3-c]cinnolin-12-ones on topoisomerase I-targeting activity and cytotoxicity was evaluated. Potent TOP1-targeting activity was observed when the 11-position was substituted with either a 2-(N,N-dimethylamino)ethyl, a 2-(N,N diethylamino)ethyl, a n-butyl, or a 2-(pyrrolidin-1-yl)ethyl group. The addition of a beta-methyl group to 1a provided an analogue with dramatically reduced TOP1 targeting activity and cytotoxicity. Analogues of 1a wherein the 2-(N,N dimethylamino)ethyl group was replaced with a (2-tetrahydrofuranyl)methyl, a 2 (piperidin-1-yl)ethyl, or a 2-(4-methylpiperazin-1-yl)ethyl substituent exhibited decreased activity as TOP1-targeting agents. Replacement of the dimethoxy groups of 1a with hydrogen atoms resulted in an analogue with significantly decreased TOP1-targeting activity and cytotoxicity. Removal of both the vicinal dimethoxyl groups and the methylenedioxy moiety resulted in a complete loss of TOP1 targeting activity. The presence of a 9-nitro substituent in place of the 8,9 methylenedioxy group of 1a resulted in a decrease in relative TOP1-targeting activity and cytotoxicity. Compounds 1a and the 11-n-butyl analogue 1d were evaluated for antitumor activity in the human tumor xenograft model using athymic nude mice. The non-estrogen responsive breast tumor cell line MDA-MB-435 was used in these assays. At dose levels that approached its maximum tolerated dose, 1a proved to be effective in inhibiting tumor growth in vivo when administered orally or by ip injection. PMID- 14759741 TI - Small molecule functional discrimination of the kinases required for the microbial synthesis of threonine and isoleucine. AB - The biosynthesis of l-threonine and l-isoleucine in bacteria and in fungi requires the action of 2 amino acid kinases: aspartate kinase and homoserine kinase. Although these kinases bind similar substrates and catalyze analogous phosphotransfer chemistry, they do not show high amino acid sequence homology. We show that despite this difference, both kinases form a ternary complex consisting of enzyme- adenosine triphosphate- amino acid to accomplish phosphoryl transfer. With this similarity in mind, we set out to identify molecules that could lead to inhibitors with activity against both kinases in the pathway. We synthesized a series of aspartic acid-adenosine bisubstrate compounds separated by a variable length alkyl linker that we hypothesized could bind to these kinases. These bisubstrate compounds only inhibited the bacterial aspartate kinase. These results reveal unexpected differences in small molecule interactions among these functionally similar enzymes. PMID- 14759742 TI - In vitro antiplasmodial activities of semisynthetic N,N'-spacer-linked oligomeric ergolines. AB - Starting from three monomeric ergolines (terguride 1, festuclavine 2, pergolide 3) N,N'-spacer-linked oligomeric derivatives were prepared using different aliphatic or arylalkyl spacers. The compounds have been evaluated for their in vitro antiplasmodial activity against the chloroquine-sensitive strain poW and the chloroquine-resistant clone Dd2 of Plasmodium falciparum. Additionally, the cytotoxic effects against mouse fibroblasts (NIH 3T3) in vitro, and human hepatocytes were evaluated. All monomers displayed only a weak antiplasmodial effect, but N-1,N-1'-spacer-linked dimerization substantially enhanced their antiplasmodial activity. The best activities were observed for compounds showing a distance of six carbon atoms between two monomers, which can be obtained by aliphatic or p-xylene linkers. The N-6,N-6'-spacer-linked depropylpergolide dimer 3i exhibited the highest antiplasmodial activity of all compounds tested (IC(50) values: 0.14 and 0.13 microM against poW and Dd2, respectively). Unfortunately, it displayed toxic effects against the mouse fibroblast cell line NIH 3T3 (IC(50): 0.1+/-0.09 microM) and also against human hepatocytes at 100 microM (LDH leakage: 15.58+/-0.87 microkat/L; GSH-level: 8.15+/-0.78 nmol/10(6) cells). However, the N-1,N-1'-spacer-linked trimer of festuclavine (2f), and also the N 1,N-1'-spacer-linked tetramer of terguride (1g) possessed remarkable antiplasmodial activities (IC(50): 0.54 and 1.53 microM, respectively, against Dd2) lacking cytotoxicity. PMID- 14759743 TI - QSAR studies on psychotomimetic phenylalkylamines. AB - Quantitative Structure-Activity Relationship (QSAR) studies on a series of psychotomimetic phenylalkylamines have been made using a combination of Minimum Topological Difference (MTD) method and topological methodology. The topological indices used being a pool of distance-based topological indices. The regression analyses have shown that excellent results are obtained in multiparametric model containing MTD parameters, topological indices in that quantum chemical parameters has to be introduced. The predictive power of the proposed model is discussed on the basis of cross-validation parameters. PMID- 14759744 TI - State-of-the-art research perspectives on musculoskeletal disorder causation and control: the need for an intergraded understanding of risk. PMID- 14759745 TI - National efforts to identify research issues related to prevention of work related musculoskeletal disorders. AB - Musculoskeletal disorders (MSDs), including low back and upper extremity disorders, represent one of the greatest work-related health concerns facing industrialized nations. Recently, two national groups were charged with developing research agendas aimed at increasing our knowledge of the prevention of these disorders. The first agenda, developed by the National Institute for Occupational Safety and Health's (NIOSH) National Occupational Research Agenda (NORA) MSD team, was based on input from several hundred practitioners and safety and health experts representing industry, labor, and academia. The second agenda, developed by the National Research Council (NRC) and the Institute of Medicine's (IOM) National Panel on Musculoskeletal Disorders and the Workplace, was based on input from leading researchers in the fields of medicine, information science, and ergonomics. This paper summarizes the findings of the two groups and compares the two research agendas. PMID- 14759746 TI - Work-related musculoskeletal disorders: the epidemiologic evidence and the debate. AB - The debate about work-relatedness of musculoskeletal disorders (MSDs) reflects both confusion about epidemiologic principles and gaps in the scientific literature. The physical ergonomic features of work frequently cited as risk factors for MSDs include rapid work pace and repetitive motion, forceful exertions, non-neutral body postures, and vibration. However, some still dispute the importance of these factors, especially relative to non-occupational causes. This paper addresses the controversy with reference to a major report recently commissioned by the US Congress from the National Research Council (NRC) and Institute of Medicine (IOM) (2001). The available epidemiologic evidence is substantial, but will benefit from more longitudinal data to better evaluate gaps in knowledge concerning latency of effect, natural history, prognosis, and potential for selection bias in the form of the healthy worker effect. While objective measures may be especially useful in establishing a more secure diagnosis, subjective measures better capture patient impact. Examination techniques still do not exist that can serve as a "gold standard" for many of the symptoms that are commonly reported in workplace studies. Finally, exposure assessment has too often been limited to crude indicators, such as job title. Worker self-report, investigator observation, and direct measurement each add to understanding but the lack of standardized exposure metrics limits ability to compare findings among studies. Despite these challenges, the epidemiologic literature on work-related MSDs-in combination with extensive laboratory evidence of pathomechanisms related to work stressors-is convincing to most. The NRC/IOM report concluded, and other reviewers internationally have concurred, that the etiologic importance of occupational ergonomic stressors for the occurrence of MSDs of the low back and upper extremities has been demonstrated. PMID- 14759747 TI - Epidemiology of musculoskeletal disorders among computer users: lesson learned from the role of posture and keyboard use. AB - Reports in the scientific literature and lay press have suggested that computer users are at increased risk of upper extremity musculoskeletal disorders (MSDs). Early studies often found elevated rates of MSD outcomes among keyboard users when compared to non-users. Attention soon focused on specific aspects of keyboard work that might be responsible for the observed rate increase. In this review, the epidemiological evidence examining associations between MSD outcomes and computer user posture and keyboard use intensity (hours of computer use per day or per week) are examined. Results of epidemiological studies of posture and MSD outcomes have not been entirely consistent. Reasons for the inconsistency in results include cross-sectional study design (with possible failure to assure that measured exposure preceded health effect), imprecision of posture measures used, and difficulties involved in analyzing multiple related variables. Despite the inconsistencies, it appears from the literature that posture is an independent risk factor of modest magnitude for MSDs among computer users. It appears that lowering the height of the keyboard to or below the height of the elbow and resting the arms on the desk surface or chair armrests is associated with reduced risk of neck and shoulder MSDs. Results of epidemiological studies examining computer use (hours keying per day or per week) are more consistent than those examining posture, although some inconsistency is observed. Reasons for the inconsistency include possible selective survival bias resulting from cross-sectional study design, differences in exposure categorization, and possible interaction with other exposure variables. Overall, the literature shows that daily or weekly hours of computer use is more consistently associated with hand and arm MSDs than with neck and shoulder MSDs. PMID- 14759748 TI - Reducing the costs of work-related musculoskeletal disorders: targeting strategies to chronic disability cases. AB - Musculoskeletal disorders impose a significant direct cost burden on health care systems in the US and Canada and account for even greater indirect losses of productivity. The overall prevalence of musculoskeletal disorders is high, but a disproportionate share of costs is associated with a small number of cases with chronic pain. This is especially true for cases of occupational back pain, the single most common and costly musculoskeletal disorder in the workplace. A number of studies identify workplace characteristics associated with prolonged disability among cases of work-related back pain. These characteristics include: failure to receive job accommodations, receipt of disability benefit payments, and employment in high-risk industries or jobs that require heavy lifting. Research on the predictors of high-cost cases is limited, however, because of the lack of high-quality data and the need for a multidisciplinary approach. A new study, the Arizona State University Healthy Back Study, addresses some of these issues and promises new insights into effective strategies to reduce the proportion of high-cost claims. PMID- 14759749 TI - Linking latest knowledge of injury mechanisms and spine function to the prevention of low back disorders. AB - While several sophisticated scientific approaches have been employed to understand low back function and injury mechanisms, very few have been broadly used to develop and justify injury prevention strategies. This paper looks beyond the linked segment model, and the lessons learned from this biomechanical approach, to consider the application of more sophisticated approaches. These include modelling approaches with greater anatomical and biological fidelity, fusing the lessons learned from the areas of tissue mechanics and concepts of spine stability, together with some studies that have examined several characteristics including psychosocial, physiological and personal variables. The objective is to better link recently discovered mechanisms of injury and spine tissue health with injury risk reducing approaches. PMID- 14759750 TI - Ligaments: a source of work-related musculoskeletal disorders. AB - The mechanical and neurological properties of ligaments are reviewed and updated with recent development from the perspective which evaluates their role as a source of neuromusculoskeletal disorders resulting from exposure to occupational activities. Creep, tension-relaxation, hysteresis, sensitivity to strain rate and strain/load frequency were shown to result not only in mechanical functional degradation but also in the development of sensory-motor disorders with short- and long-term implication on function and disability. The recently exposed relationships between collagen fibers, applied mechanical stimuli, tissue microdamage, acute and chronic inflammation and neuromuscular disorders is delineated with special reference to occupational stressors. PMID- 14759751 TI - Factors involved in strain-induced injury in skeletal muscles and outcomes of prolonged exposures. AB - Repetitive motion disorders can involve lengthening of skeletal muscles to perform braking actions to decelerate limbs under load often resulting in muscle strains and injury. Injury is a loss of isometric force (weakness) requiring days to recover. The capacity of skeletal muscle to tolerate repeated strains is dependent on multiple factors including individual variation. The most important factors producing muscle strain injury are the magnitude of the resisting force (peak-stretch force) and the number of strains. Other factors such as muscle length and fiber type contribute to the susceptibility to injury as well, but to a lesser degree. Strain injury can also lead to inflammation and pain. Chronic exposure to repeated strains can result in fibrosis that is not completely reversed after months of rest. Long rest times appear to be the only factor reported to prevent inflammation in rats following repeated strain injury. Further understanding of the mechanism for prevention of histopathologic changes by long rest times should provide a rationale for prevention of negative outcomes. PMID- 14759752 TI - Entrapment neuropathies: pathophysiology and pathogenesis. AB - A number of theories of pathogenesis of entrapment neuropathy, due to repeated loading, have been proposed and these theories are being actively explored with animal models. Tubes placed loosely around peripheral nerves cause delayed onset, chronic pain and changes in nerve morphology including nerve sprouting. Balloons placed around or adjacent to the nerve and inflated to low pressures, rapidly produce endoneurial edema and a persistent increase in intraneural pressure. The same models demonstrate long-term changes such as demyelination and fibrosis. The applied pressure causes a decrement in nerve function and abnormal morphology in a dose-dependent manner that appears to be linked to the amount of endoneurial edema. A new model involving involuntary, repetitive fingertip loading for 6 h per week for 4 weeks has caused slowing of nerve function at the wrist similar to that seen in patients with carpal tunnel syndrome. These models have the potential to reveal the mechanisms of injury at the cellular and biochemical level and address questions about the relative importance of various biomechanical factors (e.g. peak force, mean force, force rate, duty cycle, etc.). In addition, these models will allow us to evaluate various prevention, treatment and rehabilitation protocols. PMID- 14759753 TI - Inflammation reduces physiological tissue tolerance in the development of work related musculoskeletal disorders. AB - Work-related musculoskeletal disorders (MSDs) cause substantial worker discomfort, disability and loss of productivity. Due to the difficulty in analyzing the tissues of patients in the early stages of work-related MSD, there is controversy concerning the pathomechanisms of the development of these disorders. The pathophysiology of work-related MSD can be studied more easily in animal models. The purpose of this review is to relate theories of the development of tissue injury due to repeated motion to findings of recent investigations in animals that address the role of the inflammatory response in propagating tissue injury and contributing to chronic or recurring tissue injury. These tissue effects are related to behavioral indicators of discomfort and movement dysfunction with the aim of clarifying key time points for specific intervention approaches. The results from animal models of MSD are discussed in the light of findings in patients, whose tissues are examined at a much later phase of MSD development. Finally, a conceptual model of the potentially negative impact of inflammation on tissue tolerance is proposed along with suggestions for future research directions. PMID- 14759754 TI - Mechanisms of central sensitization, neuroimmunology & injury biomechanics in persistent pain: implications for musculoskeletal disorders. AB - This review will offer an overview of the mechanistic pathways of chronic pain associated with musculoskeletal disorders (MSDs). Traditional electrophysiological pain pathways of these injuries will be reviewed. In addition, recent research efforts in persistent pain have characterized a cascade of neuroimmunologic events in the central nervous system that manifests in pain behaviors and neurochemical nociceptive responses. Physiologic changes in the central nervous system will be covered as they pertain to the interplay of these two areas, and also as they focus on MSDs and injuries. One such injury leading to persistent pain is radiculopathy, which results from nerve root compression or impingement and leads to low back pain. This painful syndrome will be used as an example to provide a context for presenting immune mechanisms of chronic pain and their relationship to injury. Measures of injury biomechanics are presented in the context of the resulting pain responses, including behavioral sensitivity, local structural changes, and cellular and molecular changes in the CNS. Lastly, based on these findings and others, a discussion is provided highlighting areas of future work to help elucidate methods of injury diagnosis and development of therapeutic treatments. PMID- 14759755 TI - Review of enigmatic MTrPs as a common cause of enigmatic musculoskeletal pain and dysfunction. AB - This article explores how myofascial trigger points (MTrPs) may relate to musculoskeletal dysfunction (MSD) in the workplace and what might be done about it. The cause of much MSD and pain is often enigmatic to modern medicine and very costly, just as the cause of MTrPs has been elusive for the past century, despite an extensive literature that is confusing because of restricted regional approaches and a seemingly endless variety of names. MTrPs are activated by acute or persistent muscle overload, which is characteristic of MSD in the workplace. MTrPs can involve any, and sometimes many, of the skeletal muscles in the body and are a major, complex cause of musculoskeletal pain. The clinical and etiological characteristics of MTrPs have been underexplored by investigators, leading to undertraining of health care professionals, underappreciation of their clinical importance. MTrPs have no gold standard diagnostic criterion, and no routinely available laboratory or imaging test. MTrPs require a specific non routine examination and muscle-specific treatment for prompt relief when acute, and also resolution of perpetuating factors when chronic. After identifying a critical false assumption, electrodiagnostic studies are now making encouraging progress toward clarifying the etiology of MTrPs based on the 5- or 6-step positive-feedback model of the integrated hypothesis. Specific research needs are noted. MTrPs are treatable and they deserve increased attention and consideration by research investigators and clinicians. PMID- 14759756 TI - Biomechanics of musculoskeletal pain: dynamics of the neuromatrix. AB - Pain, due to mechanical stimuli, is a normal, indeed healthy, response of animals to potential or actual damage to tissues. Mammals in general, and humans in particular, have evolved a highly sophisticated system of pain perception, which is characterized in humans by complementary but distinct neural processing of the intensity and location of a noxious stimulus, and a motivational/emotional or affective response to the stimulus. The peripheral and central neurons that comprise this system, which has been called the 'neuromatrix', dynamically (temporally) respond and adapt to noxious biomechanical stimuli. However, phenotypic variability of the neuromatrix can be large, which can result in a host of musculoskeletal conditions that are characterized by altered pain perception, which can and often does alter the course of the condition. This neural plasticity has been well recognized in the central nervous system, but it has only more recently become known that peripheral nociceptors also adapt to their altered extracellular matrix environment. This work reviews the biomechanics of pain focusing on the relevant stimulus that initiates responses by nociceptors to the cognitive perception of pain. PMID- 14759757 TI - Individual factors and musculoskeletal disorders: a framework for their consideration. AB - Individual factors have been variously defined as non-work, demographic, physiological or psychological factors. They may represent a variety of important constructs at different relevant levels that may not be initially evident in their measurement. These include: work-related factors e.g., job assignment, duration of exposure, work style, anthropometric mismatches, and differential responses to job demands; concomitant external or internal exposures e.g., sports, smoking, and endogenous hormones; and physical, psychological and social vulnerabilities e.g., prior injury, depression, socio-economic status. Such factors operate in different ways in the development, course and response to interventions of musculoskeletal disorders. Newer framings of their contribution to musculoskeletal disorders are providing new insights into the role of such factors as some among many which contribute to the burden of MSK disorders in working age populations. As researchers, practitioners and policy makers, we need to consider them in order to reduce burden, to protect the vulnerable and to match interventions to different groups of people most appropriately. PMID- 14759758 TI - Back pain--individual and genetic factors. AB - BACKGROUND: In research, back pain can be classified in several ways. This presentation will deal with 'self-reported back pain', which is a proxy measure of 'having back pain'. The terms 'individual factors' cover many aspects of human life. They also interact with external factors, such as work and environment. Back pain is prevalent at all ages, although the incidence (onset) probably is highest in the early teens. Causative and modifying factors may differ throughout life and it is not sure that the etiology of 'back pain' remains the same. If we are to prevent and treat back pain in an efficient way, it is essential that we understand the circumstances that bring it on or prevent it from getting better. The mode of action would depend on whether the responsibility lies mainly with the individual, for example through a change of life-style, or with others, for example by altering the work place. METHOD: My presentation will consist of three elements, evidence obtained from: (1). systematic literature reviews, (2). epidemiologic studies, and (3). genetic epidemiologic studies. All the information presented is based on own work and pertains to children/adolescents and adults, mostly in relation to low back pain (LBP). The evidence will be reviewed in the light of 'the weight of evidence' on the topics of: genetics, sedentary life-style, type of work and imaging findings. Also factors relating to social class and physical/psychological robustness will be discussed. RESULTS: There is evidence for a relatively strong genetic component to LBP, both in the young and in adults. Hard work is strongly linked to LBP and there is a moderate link with abnormal imaging findings (spinal pathology in the young and discal degeneration in both the young and the adults). In addition, a weak psychological and psychological constitution appears to be of interest, at least in the young. However, there is no evidence for a causal link between a sedentary life-style and LBP, neither in the young nor in the adults. CONCLUSIONS: Some myths in the area of back pain are dispelled by the evidence in relation to personal and genetic factors and some new elements need considering. It is suggested that we should look for populations at risk, rather than looking for risk factors. Populations at risk would consist of people with a weak psychological and physiological constitution, who more easily than others may develop long lasting back pain as well as other CLINICAL SIGNIFICANCE: Early identification of high risk populations will allow for a selective primary and secondary preventive approach. PMID- 14759759 TI - Interventions to reduce work-related musculoskeletal disorders. AB - Work-related Musculoskeletal Disorders (WMSDs) continue to present a major challenge to workers and their employers in virtually every industry sector. Many disciplines have been involved in providing advice and working on interventions to prevent WMSDs or reduce their consequences. Since the early 1990s, 15 systematic reviews (excluding specific treatment modalities) have appeared in the peer-reviewed literature addressing musculoskeletal disorder reduction. The National Research Council-Institute of Medicine summarized many of these efforts in 2001. Using a systematic literature search strategy, we identified 20 randomized controlled studies, 17 quasi-experimental studies with control groups, and 36 paper reporting case studies with in the peer-reviewed literature between 1999-2003. Evidence is continuing to build that demonstrates combinations of measures appear to have the greatest effect in reducing WMSDs, although individual engineering and administrative controls can also have positive effects. PMID- 14759760 TI - Work-related low back pain: secondary intervention. AB - A review of the literature indicates that non-specific low back pain is basically an age-related disorder that is affected by differences in occupation, genetics and personal behavior. The cause of low back pain is unknown in most cases, and most of the treatments in routine use are ineffective. There is no evidence that low back pain has decreased in recent years. Consequently, there are a growing number of investigators who believe that efforts at preventing low back pain are futile; that low back pain is an unavoidable consequence of life that will afflict two thirds of the adult population at some point in their lives. These investigators believe that programs aimed at reducing low back disability are likely to be more effective and less costly. Only a small percentage of people with low back pain become disabled, but this small percentage consumes most of the cost. The literature pertaining to the reduction of low back disability is reviewed and discussed. It is concluded that low back disability can be reduced, and that management (not the health care provider) has the primary responsibility for reducing disability. PMID- 14759761 TI - Musculoskeletal disorders: primary and secondary interventions. AB - In the United States alone, the annual cost associated with the diagnosis and care of musculoskeletal trauma amounts to tens of billions of dollars [Occupational Musculoskeletal Disorders: Function, Outcomes and Evidence. Lippincott Williams and Wilkins, Philadelphia]. Moreover, these costs are continuing to increase at an alarming rate. In fact, in the United States today, occupational musculoskeletal disorders are the leading causes of work disability. Changes in health care policy and demand for improved allocation of health care resources by the Federal government have also recently placed greater pressure on health care professionals to provide the most cost-effective treatment for these disorders, as well as to validate treatment effectiveness. Indeed, treatment outcome monitoring has assumed new importance in medicine. It is particularly essential in musculoskeletal care, which is currently targeted for attention by health care planners because of its high cost and perceived traditional inefficient care. With these facts in mind, the purpose of the present article is to review the status of current primary and secondary interventions for musculoskeletal disorders. Before doing so, a brief discussion of the biopsychosocial model of pain and disability, which is currently the most heuristic approach to intervention, will be provided. PMID- 14759762 TI - From confounders to suspected risk factors: psychosocial factors and work-related upper extremity disorders. AB - Psychosocial variables have recently been more prominent among epidemiologic risk factors for work-related upper extremity disorders (WRUEDs), but bio-behavioral mechanisms underlying these associations have been elusive. One reason is that the psychosocial domain has included many broad and disparate variables (e.g. mood, coping skills, job control, job satisfaction, job stress, social support), and this lack of specificity in the conceptualization of psychosocial factors has produced limited hypothesis testing opportunities. Therefore, recent research efforts have focused on identifying and conceptualizing specific psychosocial factors that might more clearly delineate plausible bio-behavioral mechanisms linking psychosocial factors to WRUEDs. One such factor is workstyle, a strategy that workers may employ for completing, responding to, or coping with job demands that might affect musculoskeletal health. Preliminary studies have provided support for measurable differences in workstyle among individual workers and an association with upper extremity pain and discomfort. An initial self-report measure of workstyle has been pilot tested among office workers and shown acceptable reliability and validity. Future studies are needed to study this construct among other working populations and to determine its relationship with other clinical endpoints. Nevertheless, early findings suggest workstyle may be a potential focus of WRUED prevention efforts. PMID- 14759763 TI - A new frontier in cell biology: nanocellbiology. PMID- 14759764 TI - Regulation of the water channel aquaporin-1: isolation and reconstitution of the regulatory complex. AB - Aquaporins (AQP) are involved in rapid and active gating of water across biological membranes. The molecular regulation of AQP is unknown. Here we report the isolation, identification and reconstitution of the regulatory complex of AQP 1. AQP-1 and Galphai3 have been implicated in GTP-induced gating of water in zymogen granules (ZG), the secretory vesicles in exocrine pancreas. In the present study, detergent-solubilized ZGs immunoprecipitated with monoclonal AQP-1 antibody, co-isolates AQP-1, PLA2, Galphai3, potassium channel IRK-8, and the chloride channel ClC-2. Exposure of ZGs to either the potassium channel blocker glyburide, or the PLA2 inhibitor ONO-RS-082, blocked GTP-induced ZG swelling. RBC known to possess AQP-1 at the plasma membrane, swell on exposure to the Galphai agonist mastoparan, and respond similarly to ONO-RS-082 and glyburide, as ZGs. Liposomes reconstituted with the AQP-1 immunoisolated complex from solubilized ZG, also swell in response to GTP. Glyburide or ONO-RS-082 abolished the GTP effect. Immunoisolate-reconstituted planar lipid bilayers demonstrate conductance, which is sensitive to glyburide and an AQP-1 specific antibody. Our results demonstrate a Galphai3-PLA2 mediated pathway and potassium channel involvement in AQP-1 regulation. PMID- 14759765 TI - Calcium drives fusion of SNARE-apposed bilayers. AB - N-ethylmalemide-sensitive factor attachment protein receptor (SNARE) has been proposed to play a critical role in the membrane fusion process. The SNARE complex was suggested to be the minimal fusion machinery. However, there is mounting evidence for a major role of calcium in membrane fusion. Hence, the role of calcium in SNARE-induced membrane fusion was the focus of this study. It revealed that recombinant v-SNARE and t-SNARE, reconstituted into separate liposomes, interact to bring lipid vesicles into close proximity, enabling calcium to drive fusion of opposing bilayers. Exposure to calcium triggered vesicle fusion at both, high potency and efficacy. The half-time for calcium induced fusion of SNARE-reconstituted vesicles was determined to be approximately 10 s, which is two orders of magnitude faster than in its absence. Calcium acts downstream of SNAREs, since the presence of SNAREs in bilayers increases the potency of calcium-induced vesicle fusion, without significantly influencing its efficacy. Hence, this study suggests that in the physiological state in cells, both SNAREs and calcium operate as the minimal fusion machinery. PMID- 14759766 TI - C-reactive protein increases F-actin assembly and cortical distribution with resultant loss of lamellipod formation in human neutrophils. AB - C-reactive protein (CRP) inhibits neutrophil movement through a p38 MAP kinase pathway. We hypothesized that CRP altered F-actin content and distribution on human neutrophils as a means of inhibiting movement. CRP produced simultaneous increased F-actin and decreased G-actin levels. CRP increased F-actin levels in a concentration-dependent manner once a threshold (>100 microg/ml) was reached, and transiently increased F-actin (peak levels at 2.5 and 10 min) that returned to baseline by 30 min. Confocal microscopy of neutrophils revealed that fMLP provoked acquisition of a migratory phenotype as evidenced by the appearance of F actin rich lamellipods. In contrast, CRP caused neutrophil rounding, prevented lamellipod formation and shifted F-actin from the cytoskeleton to the cortex. The p38 MAP kinase inhibitor, SB203580, produced a similar effect on neutrophil shape. Concentrations of SB203580 that dramatically decreased p38 activity in neutrophils also caused round cell morphology and cortical F-actin distribution. Since CRP inhibits p38 MAP kinase and p38 blockade leads to actin polymerization and prevention of lamellipod formation, it is concluded that round morphology and loss of lamellipod formation result from CRP inhibition of p38 MAP kinase. Understanding the signal transduction of CRP prevention of lamellipod formation will aid in the development of therapeutic agents against neutrophil-associated inflammatory disease. PMID- 14759767 TI - Cell cycle and behaviour in Euplotes crassus: an attempt at describing the biological relationship between its cellular and organismic natures. AB - The two-faced nature of protozoa allowed us to study the relationship between cell cycle and behaviour in Euplotes crassus; the former represents one of the most typical cellular traits, the latter is one of the most significant characteristics of an organism. Dividing cells of E. crassus were isolated on a slide and recorded for 11 h: the classic ethographic parameters were calculated every 60 min. The percentage of mobile cells went from 0-100% in the first 2.5 h. This value was maintained for 6.5 h, but from 9 h the value began to drop, reaching 0% at 11 h. The relative frequency of leftward arcs was very high in the first hour, the radius and length of the arcs increased from 1-7 h; velocity showed a similar trend, peaking at 7 h. All our results showed that the behaviour of this ciliate is heavily affected by its cell-division cycle. PMID- 14759768 TI - Subcellular localization of MAK-V/Hunk protein kinase expressed in COS-1 cells. AB - MAK-V/Hunk is a MARK/Par-1-related protein kinase, whose function is unknown. We studied the subcellular localization of MAK-V/Hunk in COS-1 cells by immunofluorescence. It has a nucleocytoplasmic distribution and is localized to the centrosome, as indicated by co-localization with gamma-tubulin. A putative kinase-deficient mutant, with a mutation in the invariant lysine residue in the catalytic domain, was not targeted to the nucleus or centrosome. These results suggest that the nuclear and centrosomal targeting of MAK-V/Hunk is specific, and is likely to be coupled to its catalytic activity. PMID- 14759769 TI - Yessotoxin affects fMLP-induced cell shape changes inMytilus galloprovincialis immunocytes. AB - Using computer-assisted microscopic image analysis, we have found that algal yessotoxin (YTX) affects the immune response of Mytilus galloprovincialis. Indeed, YTX increases immunocyte cell motility through the involvement of both extracellular Ca2+ and cAMP, but not through protein kinase A, protein kinase C or phosphoinositide 3-kinase. Alone, however, the toxin does not induce any effect, as its action on cell motility is observed only after addition of the chemotactic substance N-formyl-Meth-Leu-Phe (fMLP). fMLP is known to induce cellular changes via both the phosphatidylinositol and cAMP pathways and, from this scenario, we can surmise that Ca2+ and cAMP concentrations rise sufficiently in fMLP-activated immunocytes to reveal YTX action. One possible explanation is that the toxin increases fMLP-mediated cell activation by intervening in L-type Ca2+-channel opening through a cAMP-dependent/PKA-independent pathway. PMID- 14759770 TI - Muscarinic acetylcholine receptors involved in the regulation of neural stem cell proliferation and differentiation in vitro. AB - Neural stem cells (NSCs) are currently considered powerful candidates for cell therapy in neurodegenerative disorders such as Parkinson's disease. However, it is not known when and how NSCs begin to differentiate functionally. Recent reports suggest that classical neurotransmitters such as acetylcholine (Ach) are involved in the proliferation and differentiation of neural progenitor cells, suggesting that neurotransmitters play an important regulatory role in development of the central nervous system (CNS). We have shown by calcium imaging and immunochemistry that proliferation and differentiation are enhanced by M2 muscarinic Ach receptors (mAchR) expressed on the NSC surface and on their neural progeny. Moreover, atropine, an mAchR antagonist, blocks the enhancement and inhibits the subsequent differentiation of NSCs. Further understanding of this neural-nutrition role of Ach might elucidate fetal brain development, the brain's response to injury, and learning and memory. PMID- 14759772 TI - Testicular pain after inguinal hernia repair: an approach to resection of the genital branch of genitofemoral nerve. AB - BACKGROUND: The neuropathic groin pain after inguinal hernia repair is usually due to a neuroma of the ilioinguinal, iliohypogastric, or genitofemoral nerve. When the postherniorrhaphy pain symptoms include mostly testicular pain, then the genital branch of the genitofemoral nerve comes first in the differential diagnosis. Nerve blocks are helpful in determining which of the three nerves is implicated in the pain syndrome. Although the surgical approach to the ilioinguinal nerve is now well established, it has been difficult to identify the genitofemoral reliably enough to permit resection of this nerve. STUDY DESIGN: Anatomic cadaver dissections of nerves exiting the lumbosacral plexus were performed. Their course was followed to their final terminations. Based on these findings, an operative approach was designed to address the involved nerve at a specific site. The results and outcomes were prospectively followed. RESULTS: The present anatomic study identifies the site within the inguinal canal where the genital branch of the genitofemoral nerve may be identified. The designed operative approach points to the proximal site of the canal to be opened for an exposure. It also dictates that the nerve should be dissected and resected proximal to the surgical repair or mesh reconstruction, which allows its retroperitoneal placement. All four patients were relieved of their preoperative symptoms using this approach. CONCLUSIONS: Severe and chronic testicular pain after inguinal hernia repair can be treated by a designed approach that identifies the genital branch of the genitofemoral nerve in the proximal inguinal canal, its resection point proximal to the previous operative field, and placement behind the peritoneum. PMID- 14759771 TI - N-acetyl-GLP-1: a DPP IV-resistant analogue of glucagon-like peptide-1 (GLP-1) with improved effects on pancreatic beta-cell-associated gene expression. AB - Glucagon-like peptide-1(7-36)amide (GLP-1) is a key insulinotropic hormone with the reported potential to differentiate non-insulin secreting cells into insulin secreting cells. The short biological half-life of GLP-1 after cleavage by dipeptidylpeptidase IV (DPP IV) to GLP-1(9-36)amide is a major therapeutic drawback. Several GLP-1 analogues have been developed with improved stability and insulinotropic action. In this study, the N-terminally modified GLP-1 analogue, N acetyl-GLP-1, was shown to be completely resistant to DPP IV, unlike native GLP 1, which was rapidly degraded. Furthermore, culture of pancreatic ductal ARIP cells for 72 h with N-acetyl-GLP-1 indicated a greater ability to induce pancreatic beta-cell-associated gene expression, including insulin and glucokinase. Further investigation of the effects of stable GLP-1 analogues on beta-cell differentiation is required to assess their potential in diabetic therapy. PMID- 14759773 TI - The clinical significance and anatomic distribution of parathyroid double adenomas. AB - BACKGROUND: Parathyroid double adenomas are reported to occur in 3% to 12% of cases of primary hyperparathyroidism, but the very existence of double adenomas has been controversial. This study was undertaken to evaluate the clinical significance and anatomic distribution of parathyroid double adenomas. STUDY DESIGN: The medical records of 384 consecutive patients who underwent operation for primary hyperparathyroidism were reviewed. RESULTS: A total of 27 patients (7%) were found to have double parathyroid adenomas. Intraoperative parathyroid hormone (PTH) levels were measured in each case. Two enlarged hypercellular parathyroid glands were identified in 6 possible configurations: 10 both superior, 3 both inferior, 5 both right, 3 both left, 5 right superior and left inferior, and 1 left superior and right inferior. There was preferential distribution to the bilateral superior position (p = 0.008). In all patients intraoperative PTH levels dropped by at least 50% from baseline and into the normal range after removal of both abnormal parathyroid glands. All patients remain normocalcemic 1 to 26 months postoperatively. Two patients have persistently elevated PTH values with normal serum calcium levels. CONCLUSIONS: The drop in intraoperative PTH levels and maintenance of normocalcemia postoperatively confirm previous reports that double adenomas do exist and are not simply missed cases of four-gland hyperplasia. Their incidence is more than would be expected by chance alone. The preferential occurrence of bilateral superior double adenomas suggests the possibility that these may represent hyperplasia of parathyroids arising from the fourth branchial pouch rather than isolated neoplastic events. PMID- 14759774 TI - Magnetic resonance imaging for preoperative evaluation of breast cancer: a comparative study with mammography and ultrasonography. AB - BACKGROUND: The widespread use of mammographic screening has led to increased detection of small tumors that are often difficult to diagnose with conventional imaging modalities such as mammography and ultrasonography. Intraductal spread of breast cancer, a principle risk factor for local recurrence, is also difficult to diagnose with mammography and ultrasonography. We investigated the clinical usefulness of magnetic resonance imaging of the breast in the therapy of breast cancer and we compared it with mammography and ultrasonography. STUDY DESIGN: A total of 183 patients with primary breast cancer underwent surgery at our institute between September 1, 1999, and November 30, 2002. They were examined preoperatively with magnetic resonance imaging, mammography, and ultrasonography. Magnetic resonance imaging evaluation included contrast-enhanced dynamic studies using IV injection of gadolinium-diethylenetriamine pentaacetic acid. RESULTS: Detection rates of breast cancers by magnetic resonance imaging, mammography, and ultrasonography were 93.7%, 84.6%, and 97.3%, respectively (magnetic resonance imaging versus mammography, p < 0.05). Patterns of time-intensity curves in dynamic magnetic resonance imaging differed with histologic types. Sensitivity, specificity, and accuracy of detection of intraductal spread were 66.7%, 64.2%, and 65.6% with MRI; 22.2%, 85.7%, and 50% with mammography; and 20.6%, 85.2%, and 50% with ultrasonography, respectively (sensitivity, specificity, and accuracy; p < 0.05, respectively). CONCLUSIONS: Magnetic resonance imaging can diagnose breast cancer as accurately as ultrasonography and more accurately than mammography. Patterns of time-intensity curves correlated with tumor histology. In addition, magnetic resonance imaging can detect intraductal spread more accurately than the other two methods. Magnetic resonance imaging appears to be indispensable in breast-conserving surgery to minimize local recurrence. PMID- 14759775 TI - Treatment of hypophosphatemia using a protocol based on patient weight and serum phosphorus level in a surgical intensive care unit. AB - BACKGROUND: Hypophosphatemia may cause organ derangements in the surgical intensive care unit. The purpose of this study was to determine the impact of a repletion protocol for hypophosphatemia based on admission weight and phosphorus level. STUDY DESIGN: All patients who presented to an 18-bed surgical intensive care unit with a serum phosphorus level of 2.2 mg/dL or less or who received phosphorus supplementation despite having normal levels were identified. In the preintervention phase between January and June 2001, 137 patients were retrospectively identified who met these criteria. A protocol was then designed giving a single intravenous dose of phosphorus based on weight and serum phosphorus. Repletion was given with sodium or potassium phosphorus based on presupplementation levels. After protocol implementation 141 patients met these criteria between September 2001 and February 2002, and treatment and postrepletion levels were followed prospectively. RESULTS: A total of 47 patients were repleted before the intervention with adequate followup and 22 (47%) attained a normal level. Supplementation success was 53% in moderate hypophosphatemia (2.2 mg/dL or less) and 27% in severe hypophosphatemia (less than 1.5 mg/dL). After protocol implementation, 111 patients were repleted with 84 (76%) correcting to a normal level (p = 0.002 compared with retrospective patients). Success was 78% in moderate hypophosphatemia and 62% in severe hypophosphatemia. Inappropriate supplementation of normal phosphorus levels decreased from 51 to 16 patients after protocol implementation. CONCLUSIONS: A protocol based on weight and serum levels successfully treated both moderate and severe hypophosphatemia in the majority of critically ill patients. Protocol implementation also decreased unnecessary supplementation of normal phosphorus levels. PMID- 14759776 TI - Recurrence pattern of squamous cell carcinoma of the thoracic esophagus after extended radical esophagectomy with three-field lymphadenectomy. AB - BACKGROUND: Extended radical esophagectomy with three-field lymphadenectomy for patients with thoracic esophageal cancer has been shown to be effective. But even if this operation is performed, some patients still experience relapse of the disease. The purpose of this study was to clarify the pattern and timing of recurrence after extended radical esophagectomy. STUDY DESIGN: Recurrence of esophageal squamous cell carcinoma was examined in 171 of 174 patients who underwent extended radical esophagectomy with three-field lymphadenectomy. Recurrence patterns were classified as locoregional (at the site of the primary tumor, the anastomotic site, or the lymph nodes), hematogenous, and other (pleura or site of gastrostomy). Factors associated with recurrence were identified using univariate and multivariate statistical methods for survival analysis. RESULTS: The overall 5-year survival rate was 55.6%. Recurrence was recognized in 74 patients (43.3%). The median disease-free interval until recurrence was 11 months. Thirty patients (17.5%) developed a locoregional recurrence, and 24 (14.0%) developed a hematogenous recurrence. Five patients (2.9%) developed both recurrences simultaneously and were classified as hematogenous recurrences. Of 30 patients with cervical lymph node metastasis, recurrent disease was recognized in 19 patients (63.3%). In multivariate analysis of 160 patients, the depth of invasion and pM-lym (cervical or celiac lymph node metastasis) were significant factors for locoregional recurrence; the depth of invasion and number of lymph node metastases at operation were significant factors for hematogenous recurrence. Survival time for patients with hematogenous recurrence (median 16 months) was significantly shorter than that of patients with locoregional recurrence (median 25.5 months). CONCLUSIONS: Locoregional recurrence is associated mainly with the extent of the local tumor and lymph node metastasis; hematogenous recurrence is not only associated with tumor stage but also with the tumor's oncologic behavior. PMID- 14759777 TI - Effect of semifundoplication with subtotal gastrectomy for prevention of postoperative gastroesophageal reflux. AB - BACKGROUND: Postgastrectomy patients often experience reflux esophagitis and a compromised quality of life. We hypothesized that reconstructive methods with antireflux procedures at operation should prevent reflux esophagitis and improve the likelihood of a better quality of life in patients after distal gastrectomy for gastric carcinoma. Our antireflux procedure was a subdiaphragmatic semifundoplication. We aim to substantiate, with objective arguments, potential advantages of Billroth I simple reconstruction versus Billroth I with semifundoplication. STUDY DESIGN: This study evaluated 60 patients who had Billroth I reconstruction with semifundoplication (30 patients; F group) and simple Billroth I reconstruction (30 patients; B group) after distal gastrectomy for gastric cancer. Assessments were made preoperatively and 6 months or later after surgical intervention. Results of the procedure, clinical evaluation (reflux symptoms), and esophageal alkaline reflux by ambulatory 24-hour pH memory were satisfactory. RESULTS: Operative evaluation time and procedural complications did not differ significantly between the two gastrectomy groups. Reflux symptoms only occurred in 12 patients in the B group. Lower esophageal sphincter pressure of patients in the B group was significantly lower than that of patients in the F group and in preoperative states (p < 0.05). The mean appearance of alkaline esophageal reflux in the F group and the B group were 2.6% and 13.6%, respectively (p < 0.01). Patients with semifundoplication had a significantly better quality of life and less physiologic regurgitation than patients with simple Billroth I. CONCLUSIONS: This study demonstrated that Billroth I reconstruction with semifundoplication for gastric cancer is not only effective for patients with a postoperative life expectancy, but also prevents reflux esophagitis after gastrectomy. We believe that our method is an effective and simple surgical option for many patients with gastric cancer. PMID- 14759778 TI - Percutaneous management of bile duct strictures and injuries associated with laparoscopic cholecystectomy: a decade of experience. AB - BACKGROUND: The 1990s were associated with a dramatic increase in bile duct injuries with the widespread use of laparoscopic cholecystectomy (LC). Interventional radiology has an integral role in diagnosing and managing these injuries. Definitive percutaneous management with balloon dilatation might be possible in select patients with intact biliary-enteric continuity, but longterm data are limited. STUDY DESIGN: Data were collected prospectively on 51 consecutive patients with major bile duct stricture or injury associated with LC, treated with percutaneous management, January 1, 1990, to December 31, 1999. Percutaneous transhepatic cholangiography and biliary catheter placement were followed by balloon dilatation and stenting. Outcomes were assessed with direct patient contact or hospital records. RESULTS: All patients completed treatment, and 50 (98%) were stent free at mean followup of 76 months. The success rate of percutaneous management was 58.8%, without need for subsequent intervention. Presenting symptoms, level of injury, and number of stents or dilatations did not predict outcomes. Percutaneous treatment was more likely to fail in patients stented for less than 4 months (p < 0.001). Operative repair at Hopkins before percutaneous management was predictive of a successful outcome (p < 0.05). Including subsequent operations or percutaneous management, successful outcomes were achieved in 98% of patients. CONCLUSIONS: Major bile duct injuries after LC remain a clinical challenge. Although surgical reconstruction is the treatment cornerstone, selected patients with biliary-enteric continuity can achieve successful long-term results with definitive percutaneous management. The combination of percutaneous management and surgical reconstruction results in successful outcomes in virtually all patients. PMID- 14759779 TI - Additional evidence in support of withholding or terminating cardiopulmonary resuscitation for trauma patients in the field. AB - BACKGROUND: Survival for trauma patients who receive prehospital cardiopulmonary resuscitation (CPR) has been reported as poor. We assessed the survival for prehospital CPR in our trauma system and attempted to find prehospital predictors of mortality. STUDY DESIGN: We conducted a retrospective review of our Level I trauma center's database that identified 588 patients over a 6-year period (January 1, 1997, to December 31, 2002) who received prehospital CPR. Mechanisms of injury, prehospital vital signs, and survival to discharge were analyzed. RESULTS: Twenty-two of 588 patients (3.7%) survived to hospital discharge. Overall, 60.7% did not survive to achieve hospital admission, and an additional 32.6% died on the first hospital day. Patients with penetrating injuries had a significantly lower survival rate than those with either blunt or other (eg, drowning, hanging) injuries (0.9% versus 6.2%, and 13.2%, respectively, p < 0.001) and significantly lower Revised Trauma Scores (RTS; mean +/- SD: 0.32 +/- 0.96 versus 0.76 +/- 1.84 and 1.18 +/- 2.51, respectively, p < 0.05.) The likelihood of survival with RTS = 0 was less than 1% overall, and 0% for penetrating trauma. CONCLUSIONS: These findings add support to recent guidelines regarding the termination or withholding of resuscitation for trauma patients in the prehospital setting. Victims of penetrating trauma with a prehospital RTS = 0 (combination of no respiratory rate, no systolic blood pressure, and a Glasgow Coma Score of 3) should be declared "dead at the scene." PMID- 14759780 TI - Management of adult blunt splenic injuries: comparison between level I and level II trauma centers. AB - BACKGROUND: The factors important in determining outcome when managing adult blunt splenic injuries continue to be debated. Whether trauma center level designation (Level I versus Level II) affects patient management has not been evaluated. STUDY DESIGN: We conducted a retrospective analysis of prospectively gathered data from the Pennsylvania Trauma Outcome Study database that collected information from 27 statewide trauma centers (Level I [15], Level II [17]). Adult patients (ages > or = 16 years) with blunt splenic injuries (ICD-9-CM 865) were evaluated. Demographic data, injury data, and trauma center level designation were collected, and patient management, length of stay, and mortality were analyzed. RESULTS: There were 2,138 adult patients who suffered blunt splenic injuries during the study period (1998-2000). Patients treated at Level II trauma centers (n = 772) had a higher rate of operative treatment (38.2% versus 30.7%) (p < 0.001), but a shorter mean length of stay (10.1 +/- 0.4 versus 12.0 +/- 0.4 days) (p < 0.01) compared with patients in Level I trauma centers (n = 1,366). The rate of failure of nonoperative treatment was lower at Level II trauma centers (13.0% versus 17.6%) (p < 0.05), but the mortality for patients managed nonoperatively was higher (8.4% versus 4.5%) (p < 0.05). Splenorrhaphy was performed more frequently in Level I trauma centers. CONCLUSIONS: Management differences exist in the treatment of adult blunt splenic injuries between institutions of different trauma center level designation. Multicenter studies should account for this finding in design and implementation. PMID- 14759781 TI - Magnetic resonance imaging in the preoperative evaluation of breast cancer: primum non nocere. PMID- 14759782 TI - Postoperative gastroesophageal reflux. PMID- 14759783 TI - What's new in burns and metabolism. PMID- 14759785 TI - The potential role of aprotinin in the perioperative management of malignant tumors. PMID- 14759786 TI - Urogenital diaphragm: an erroneous concept casting its shadow over the sphincter urethrae and deep perineal space. PMID- 14759787 TI - Concentration versus specialization? A case study on the arteria aspera. PMID- 14759788 TI - Clinical palliative care for surgeons: part 1. PMID- 14759789 TI - Solitary fibrous tumor in the retroperitoneum. PMID- 14759790 TI - Repair of an abdominal wall defect after a salvage laparotomy for sepsis. PMID- 14759791 TI - Costs of open versus endovascular abdominal aortic aneurysm repair. PMID- 14759793 TI - Surgery for the control of morbid obesity. PMID- 14759794 TI - Right portal vein ligation. PMID- 14759798 TI - Sleeping beauty transposition: biology and applications for molecular therapy. AB - Transposable elements can be considered as natural, nonviral gene-delivery vehicles and are valuable and widely used tools for germ-line transgenesis and insertional mutagenesis in invertebrate systems such as flies and worms. Such tools were not available for genome manipulations in vertebrates until recently, when an active element was resurrected from transposon fossils found in fish genomes. This element, the Sleeping Beauty transposon, shows efficient transposition in cells of a wide range of vertebrates, including humans. Sleeping Beauty transposition is a cut-and-paste process, during which the element "jumps" from one DNA molecule to another. Transposon integration into chromosomes provides the basis for long-term, or possibly permanent, transgene expression in transgenic cells and organisms. Thus, the reconstruction of the Sleeping Beauty element generated considerable interest in developing efficient and safe vectors for vertebrate transgenesis as well as for human gene therapy. In this review we summarize our current knowledge of Sleeping Beauty biology and describe the strengths and current limitations of transposon technology for gene therapeutic applications. PMID- 14759799 TI - Efficient gene transfer to hematopoietic repopulating cells using concentrated RD114-pseudotype vectors produced by human packaging cells. PMID- 14759800 TI - In vivo selection for human and murine hematopoietic cells transduced with a therapeutic MGMT lentiviral vector that inhibits HIV replication. AB - We have developed an HIV-based lentiviral vector, VRX496, which efficiently transduces human CD34+ progenitors and CD4+ T lymphocytes. VRX496 contains an antisense sequence against the HIV envelope and is currently being evaluated for safety in a clinical trial for treatment of HIV. Selective outgrowth of transduced hematopoietic cells in vivo is anticipated to increase the therapeutic efficacy of this treatment by maximizing the persistence of virus-resistant cells in the body. Although HIV resistance is selective, additional selection may aid in treatment efficacy due to the vast quantity of target cells. Therefore, we engineered VRX496 to express the P140K MGMT gene to drive potent drug-mediated in vivo selection for transduced hematopoietic long-term repopulating cells. Suboptimally transduced T cell cultures treated with O6-benzylguanine and BCNU were selected from 3 to 100%, and after selection cultures did not support HIV replication. Primary CD34+ progenitors derived from G-CSF-mobilized peripheral blood were transduced at 27 to 35% efficiency. Approximate sixfold selection was observed for transduced CD34+ progenitors, colony-forming units, and long-term culture-initiating cells. Multilineage in vivo selection was demonstrated for transduced murine hematopoietic cells in human CD34(+)-derived hematopoietic cells in NOD-SCID mice. These results establish efficient ex vivo and in vivo selection for hematopoietic cells transduced with lentiviral vectors and support the potential therapeutic benefit of this strategy in human gene therapy. PMID- 14759801 TI - Antioxidant gene therapy can protect hearing and hair cells from ototoxicity. AB - Aminoglycosides are commonly used antibiotics that often induce ototoxicity leading to permanent hair cell loss and hearing impairment. The ototoxic effects of aminoglycosides have been linked to oxidative stress. To determine the feasibility of antioxidant gene therapy for protecting the inner ear against aminoglycoside-induced oxidative stress, we used adenoviral vectors for overexpression of catalase, Cu/Zn superoxide dismutase (SOD1), and Mn superoxide dismutase (SOD2). We inoculated adenoviruses designated Ad.cat, Ad.SOD1, and Ad.SOD2 into the left guinea pig cochlea. Five days later, an ototoxic combination of kanamycin and ethacrynic acid was systemically administered. Artificial perilymph and adenovirus without a gene cassette (Ad.null) were used as controls. Biochemical analysis showed significant increase in catalase and a moderate elevation in SOD2 levels in tissues of the cochlea inoculated with the respective vectors. Auditory brain-stem responses were measured to monitor hearing thresholds. Animals were sacrificed 7 days after the ototoxic insult and their hair cells counted. Hair cells and hearing thresholds were significantly protected by Ad.cat and Ad.SOD2, while results with Ad.SOD1 were inconsistent. Control ears showed no significant protective effects. The results demonstrate that the expression of functional enzymes in the inner ear is feasible using adenoviral-mediated gene delivery. Furthermore, they confirm that reactive oxygen species contribute to aminoglycoside ototoxicity and suggest antioxidant gene therapy as a potential therapeutic strategy to reduce inner ear oxidative stress. PMID- 14759802 TI - In utero gene therapy rescues vision in a murine model of congenital blindness. AB - The congenital retinal blindness known as Leber congenital amaurosis (LCA) can be caused by mutations in the RPE65 gene. RPE65 plays a critical role in the visual cycle that produces the photosensitive pigment rhodopsin. Recent evidence from human studies of LCA indicates that earlier rather than later intervention may be more likely to restore vision. We determined the impact of in utero delivery of the human RPE65 cDNA to retinal pigment epithelium cells in a murine model of LCA, the Rpe65(-/-) mouse, using a serotype 2 adeno-associated virus packaged within an AAV1 capsid (AAV2/1). Delivery of AAV2/1-CMV-hRPE65 to fetuses (embryonic day 14) resulted in efficient transduction of retinal pigment epithelium, restoration of visual function, and measurable rhodopsin. The results demonstrate AAV-mediated correction of the deficit and suggest that in utero retinal gene delivery may be a useful approach for treating a variety of blinding congenital retinal diseases. PMID- 14759803 TI - BDNF gene-modified mesenchymal stem cells promote functional recovery and reduce infarct size in the rat middle cerebral artery occlusion model. AB - Examination of the clinical therapeutic efficacy of using bone marrow stromal cells, including mesenchymal stem cells (MSC), has recently been the focus of much investigation. MSC were reported to ameliorate functional deficits after stroke in rats, with some of this improvement possibly resulting from the action of cytokines secreted by these cells. To enhance such cytokine effects, we transfected telomerized human MSC with the BDNF gene using a fiber-mutant F/RGD adenovirus vector and investigated whether these cells contributed to improved functional recovery in a rat transient middle cerebral artery occlusion (MCAO) model. BDNF production by MSC-BDNF cells was 23-fold greater than that seen in uninfected MSC. Rats that received MSC-BDNF showed significantly more functional recovery than did control rats following MCAO. Specifically, MRI analysis revealed that the rats in the MSC-BDNF group exhibited more significant recovery from ischemia after 7 and 14 days. The number of TUNEL-positive cells in the ischemic boundary zone was significantly smaller in animals treated with MSC-BDNF compared to animals in the control group. These data suggest that MSC transfected with the BDNF gene may be useful in the treatment of cerebral ischemia and may represent a new strategy for the treatment of stroke. PMID- 14759804 TI - Development of efficient viral vectors selective for vascular smooth muscle cells. AB - The vascular smooth muscle cell (SMC) is integral to the pathogenesis of neointimal formation associated with late vein graft failure, in-stent restenosis, and transplant arteriopathy. Viral vectors transduce SMC with low efficiency and hence, there is a need for improvement. We aimed to enhance the efficiency and selectivity of gene delivery to human SMC. Targeting ligands were identified using phage display on primary human saphenous vein SMC with linear and cyclic libraries. Two linear peptides, EYHHYNK (EYH) and GETRAPL (GET), were incorporated into the HI loop of adenovirus (Ad) fibers and the capsid protein of adeno-associated virus-2 (AAV-2). Exposure of human venous SMC to EYH-modified (but not the GET-modified) Ad vector resulted in a significant increase in transgene expression levels at short, clinically relevant exposure times. Similarly, the EYH-modified AAV vector resulted in enhanced gene transfer to human venous SMC but not endothelial cells in a time- and dose-dependent manner. The EYH-modified AAV vector also enhanced (up to 70-fold) gene delivery to primary human arterial SMC. Hence, incorporation of EYH into Ad and AAV capsids resulted in a significant and selective enhancement in transduction of SMC and has implications for improving local gene delivery to the vasculature. PMID- 14759805 TI - Lentiviral vectors efficiently transduce quiescent mature 3T3-L1 adipocytes. AB - Obesity is associated with many serious afflictions such as cardiovascular disease, cancer, and diabetes. One of the main cellular systems used to study the underlying physiological and biological processes is the 3T3-L1 preadipocyte differentiation model. However, studies on 3T3-L1 adipocytes are hampered by the fact that genetic modification of mature adipocytes is notoriously difficult. In this report, we evaluated the use of lentivirus-mediated gene transfer into 3T3 L1 mature adipocytes. We demonstrate that quiescent, fully differentiated 3T3-L1 adipocytes as well as 3T3-L1 preadipocytes can be efficiently transduced with HIV 1-derived lentiviral vectors. Upon transduction using LV-PGK-GFP lentiviral vector at 100 ng p24 per 10(5) cells, more than 95% of the 3T3-L1 adipocytes in the culture expressed the GFP reporter gene. There were no overt signs of toxicity or cytopathogenicity in the cultures. Furthermore, modification of undifferentiated preadipocytes did not affect their capacity to differentiate. In addition, insulin-induced glucose uptake was not affected by the procedure. In contrast, adenoviral-mediated gene transfer into 3T3-L1 adipocytes is associated with marked cytopathogenicity. From these data, we conclude that lentiviral vectors are the gene-transfer system of choice for genetic modification of mature adipocytes. The availability of an efficient vector system may stimulate the use of adipose tissue as a target for gene therapy in obesity and other disorders. PMID- 14759806 TI - Ablating CAR and integrin binding in adenovirus vectors reduces nontarget organ transduction and permits sustained bloodstream persistence following intraperitoneal administration. AB - To create tumor-targeted Ad vectors, ablation of native CAR and integrin receptor binding is crucial to enhance the specificity of tumor transduction. Toward this aim, we have previously created base vectors in which binding to CAR (single ablated) or to both CAR and integrins (double-ablated) has been ablated. In this study, the biodistribution of the conventional (CAR and integrin binding intact), single-ablated, and double-ablated vectors was evaluated following intraperitoneal administration. The mesothelial lining of the peritoneal organs was the principle site of CAR-dependent gene transfer by the conventional vector. Surprisingly, the single-ablated vector strongly transduced the liver parenchyma rather than the mesothelium, while the double-ablated vector did not significantly transduce the parenchyma or mesothelium. The high level of parenchymal transduction by the single-ablated vector suggested that it efficiently entered the bloodstream from the peritoneal cavity. Consistent with this hypothesis, a large proportion of active particles distributed and persisted in the bloodstream following intraperitoneal administration of either the single- or the double-ablated vector. The above results suggest that the double-ablated vector backbone may not only significantly improve targeting to cancers located in the peritoneal cavity, but may also significantly improve targeting to metastatic tumors located throughout the body by virtue of its enhanced bloodstream persistence. PMID- 14759807 TI - AAV2 vector harboring a liver-restricted promoter facilitates sustained expression of therapeutic levels of alpha-galactosidase A and the induction of immune tolerance in Fabry mice. AB - The successful application of gene therapy for the treatment of genetic diseases such as Fabry is reliant on the development of vectors that are safe and that facilitate sustained expression of therapeutic levels of the transgene product. Here, we report that intravenous administration of a recombinant AAV2 vector encoding human alpha-galactosidase A under the transcriptional control of a liver restricted enhancer/promoter (AAV2/DC190-alphagal) generated significantly higher levels of expression in BALB/c and Fabry mice than could be realized using the ubiquitous CMV promoter (AAV2/CMVHI-alphagal). Moreover, AAV2/DC190-alphagal mediated hepatic expression of alpha-galactosidase A was sustained for 12 months in BALB/c mice and was associated with a significantly reduced immune response to the expressed enzyme. Subsequent challenge of the AAV2/DC190-alphagal-treated animals with recombinant human alpha-galactosidase A at 6 months failed to elicit the production of anti-alpha-galactosidase A antibodies, suggesting the induction of immune tolerance in these animals. The levels of expression attained with AAV2/DC190-alphagal in the Fabry mice were sufficient to reduce the abnormal accumulation of globotriaosylceramide in the liver, spleen, and heart to basal levels and in the kidney by approximately 40% at 8 weeks. Together, these results demonstrate that AAV2-mediated gene transfer that limits the expression of alpha galactosidase A to the liver may be a viable strategy for treating Fabry disease. PMID- 14759808 TI - Contribution of Toll-like receptor 9 signaling to the acute inflammatory response to nonviral vectors. AB - Immunostimulatory CpG motifs have been implicated as a major contributor to the acute inflammatory response associated with nonviral vectors, most prominently seen after systemic delivery of cationic lipid-plasmid DNA (pDNA) complexes. We have shown previously that complexes containing pDNA vectors that have been largely depleted of CpG motifs have significantly reduced acute toxicity when delivered systemically. However, several CpGs remain in these vectors and the toxicity is not negligible, especially at higher doses of complex. To determine the maximal reduction in the acute toxic response that could be achieved by eliminating CpG signaling, we injected cationic lipid-pDNA complexes into transgenic mice that are deficient in Toll-like receptor 9 (TLR9), which is the receptor that recognizes immunostimulatory CpG motifs. We observed significantly decreased adverse hematological changes and liver damage in TLR9(-/-) mice compared to normal mice and increased survival at higher doses of complex. However, a pronounced loss of lymphocytes and platelets was still observed in the TLR9(-/-) mice at higher doses. We also measured the toxicity in normal mice of systemically delivered complexes containing non-CpG oligonucleotides. Although serum transaminase levels were reduced, a loss of lymphocytes and platelets akin to that seen in the TLR9(-/-) mice was observed. Taken together, these findings suggest that signaling through TLR9 contributes to the majority but not all of the toxic responses associated with systemic delivery of cationic lipid-pDNA complexes. PMID- 14759809 TI - CpG methylation of a plasmid vector results in extended transgene product expression by circumventing induction of immune responses. AB - Gene therapy has the potential to cure inherited diseases if the delivered genes achieve long-term expression at therapeutic levels in the targeted tissues. Expression is commonly short-lived due to induction of cell-mediated immune responses to the gene therapy vehicle and/or the transgene product, which can be perceived as "foreign" by the host's immune system. Plasmid expression vectors have been used to deliver genes. Bacterial DNA carries immunostimulatory sequences in the form of unmethylated CpG motifs, which induce an inflammatory reaction that in turn promotes activation of transgene product-specific B and T cells. Elimination or methylation of immunostimulatory CpG sequences in plasmid expression vectors prevents the stimulation of transgene product-specific immune responses without necessarily reducing transgene expression. In this study, we tested if a CpG-methylated plasmid expression vector expressing the highly immunogenic glycoprotein of rabies virus can achieve prolonged transgene product expression by circumventing immune recognition. Our data show that mice inoculated with a CpG-methylated plasmid expression vector show delayed clearance of transfected cells and fail to mount a strong immune response to the transgene product. Gene transfer with a CpG-methylated plasmid results in a state of immunological low responsiveness to the transgene product, which may facilitate readministration of the transgene. Nevertheless, mice remain responsive to the transgene product delivered by a viral vector. PMID- 14759810 TI - The effects of rAAV2-mediated NGF gene delivery in adult and aged rats. AB - Nerve growth factor (NGF) therapy has been proposed to treat patients with age related cognitive deficits, including those with Alzheimer's disease. One promising approach to delivering this protein into brain involves viral vectors. However, little is known about the effects of aging on gene transfer in brain generally and in particular its effect on transgenic NGF expression. To examine the transgene expression and biological effects of NGF gene transfer in adult and aged rats, we delivered mouse NGF with C-terminal myc-tag, using a recombinant adeno-associated virus serotype 2 (rAAV2) vector, into the septum of 6- and 21 month-old Fischer 344/Brown Norway hybrid rats. Other animals received a control vector encoding green fluorescent protein. As expected, this strain of rat demonstrated very few age-related deficits in spatial memory-related behavior in the Morris water task either before gene transfer (6 vs 21 months) or afterward (up to 11 vs 26 months). We found that rAAV2 vectors drove transgene expression in aged rats up to 5 months, although the level of transgene expression was lower than that of adult animals. We also showed that NGF gene transfer into the septum of aged animals induced local trophic effects by increasing the number and soma area of septal cholinergic neurons and improved distal synaptic activity by increasing the level of depolarization-induced acetylcholine (ACh) release from hippocampal synaptic terminals. Interestingly, NGF gene transfer suppressed depolarization-induced ACh release in adult rats. These findings show for the first time, to our knowledge, that septal NGF gene transfer modulates hippocampal nerve terminal function. These results are relevant for the potential clinical application of NGF gene therapy. PMID- 14759811 TI - Nerve growth factor receptor-mediated gene transfer. AB - One obstacle to effective gene therapies for neurological disorders lies in the cell-type diversity of the nervous system, making it difficult to direct gene delivery vectors to specific types of cells. To meet this challenge, we have developed a recombinant peptide-based gene delivery vector that targets nerve growth factor (NGF) receptors. The peptide comprises a cell-targeting domain derived from the NGF hairpin motif containing loops 1 and 2 linked to a DNA binding domain composed of SPKR repeats. In PC12 cells, it activated the high affinity NGF receptor, TrkA, and displayed NGF-like bioactivity by promoting neurite outgrowth and cell survival after serum deprivation. When combined with a low molecular weight of polyethylenimine (PEI), the peptide condensed plasmid DNA into nanoparticles that efficiently transferred exogenous genes into PC12 cells, enhancing reporter gene expression 5600-fold over peptide-free DNA/PEI complexes. Co-incubation with free NGF inhibited this effect. Furthermore, the peptide enhanced gene expression in NGF-receptor-rich rat primary cortex neurons but not glial cells. An in vivo experiment targeting TrkA-expressing dorsal root ganglia demonstrated that the peptide-containing complexes were 9- to 14-fold more efficient in transfection than controls. These properties make the chimeric peptide a promising gene delivery vector for targeting specific subtypes of neurons. PMID- 14759812 TI - Enhanced gene delivery by avidin-displaying baculovirus. AB - Flexible alteration of virus surface properties would be beneficial for enhanced and targeted gene delivery. A useful approach could be based on a high-affinity receptor-ligand pair, such as avidin and biotin. In this study, we have constructed an avidin-displaying baculovirus, Baavi. Avidin display was expected to enhance cell transduction due to the high positive charge of avidin in physiological pH and to provide a binding site for covering the virus with desired biotinylated ligands. Successful incorporation of avidin on the virus envelope was detected by immunoblotting and electron microscopy. Multiple biotin binding sites per virus were detected with fluorescence-correlation spectroscopy and tight biotin binding was observed using an optical biosensor, IAsys. Baavi showed a 5-fold increase in transduction efficiency in rat malignant glioma cells (BT4C) and a 26-fold increase in rabbit aortic smooth muscle (RAASMC) cells compared to wild-type baculovirus. Enhanced transduction was also observed with biotinylated target cells. Biotinylated epidermal growth factor (EGF) enabled specific targeting of the virus with high efficiency to EGF receptor-expressing (SKOV-3) cells. An additional advantage of the avidin display was demonstrated with biotinylated paramagnetic particles, which enabled magnetic targeting. Altogether, we show that avidin display is a rapid and versatile method to improve viral properties for gene delivery. PMID- 14759813 TI - Development of hyperactive sleeping beauty transposon vectors by mutational analysis. AB - The Sleeping Beauty (SB) transposable element is a promising vector for transgenesis in vertebrates and is being developed as a novel, nonviral system for gene therapeutic purposes. A mutagenesis approach was undertaken to improve various aspects of the transposon, including safety and overall efficiency of gene transfer in human cells. Deletional analysis of transposon sequences within first-generation SB vectors showed that the inverted repeats of the element are necessary and sufficient to mediate high-efficiency transposition. We constructed a "sandwich" transposon, in which the DNA to be mobilized is flanked by two complete SB elements arranged in an inverted orientation. The sandwich element has superior ability to transpose >10-kb transgenes, thereby extending the cloning capacity of SB-based vectors. We derived hyperactive versions of the SB transposase by single-amino-acid substitutions. These mutations act synergistically and result in an almost fourfold enhancement of activity compared to the wild-type transposase. When combined with hyperactive transposons and transiently overexpressed HMGB1, a cellular cofactor of SB transposition, hyperactive transposases elevate transposition by almost an order of magnitude compared to the first-generation transposon system. The improved vector system should prove useful for efficient gene transfer in vertebrates. PMID- 14759814 TI - Model of unidirectional transluminal gene transfer. AB - Gene transfer assays in vitro are poor indicators of transduction efficacy observed in vivo. We designed and optimized an intermediate model for assessing and quantifying unidirectional transduction ex vivo. The model enables simultaneous transmucosal evaluation of up to 96 different variables under the same tissue conditions. We show that the model is versatile and suitable for use with different vectors (adenovirus and AAV), different reporter genes (beta galactosidase and green fluorescent protein), and viscera with various tissue features such as peritoneum and urothelium. Ex vivo transduction assays may correlate better with in vivo gene transfer results. Because the experimental model described here can be performed in small samples, it may enable translational applications in tissues of human origin. PMID- 14759815 TI - Research on structural issues of the UMLS--past, present, and future. PMID- 14759816 TI - Exploring semantic groups through visual approaches. AB - Objectives. We investigate several visual approaches for exploring semantic groups, a grouping of semantic types from the Unified Medical Language System (UMLS) semantic network. We are particularly interested in the semantic coherence of the groups, and we use the semantic relationships as important indicators of that coherence. Methods. First, we create a radial representation of the number of relationships among the groups, generating a profile for each semantic group. Second, we show that, in our partition, the relationships are organized around a limited number of pivot groups and that partitions created at random do not exhibit this property. Finally, we use correspondence analysis to visualize groupings resulting from the association between semantic types and the relationships. Results. The three approaches provide different views on the semantic groups and help detect potential inconsistencies. They make outliers immediately apparent, and, thus, serve as a tool for auditing and validating both the semantic network and the semantic groups. PMID- 14759817 TI - Designing metaschemas for the UMLS enriched semantic network. AB - The enriched semantic network (ESN) has previously been presented as an enhancement of the semantic network (SN) of the UMLS. The ESN's hierarchy is a DAG (Directed Acyclic Graph) structure allowing for multiple parents. The ESN is thus more complex than the SN and can be more difficult to view and comprehend. We have previously introduced the notion of a metaschema for the SN as a compact abstraction to support SN comprehension. We extend the definition of metaschema to make it applicable to a DAG classification hierarchy, such as the one exhibited by the ESN. We specify the requirements for and describe the general process of deriving such a metaschema. We derive two particular metaschemas of the ESN based on a pair of partitions. These two metaschemas and their underlying partitions are compared. Both metaschemas serve as compact representations of the ESN, allowing for convenient viewing of its hierarchy and easier comprehension. PMID- 14759818 TI - Consistency across the hierarchies of the UMLS Semantic Network and Metathesaurus. AB - OBJECTIVE: To develop and test a method for automatically detecting inconsistencies between the parent-child is-a relationships in the Metathesaurus and the ancestor-descendant relationships in the Semantic Network of the Unified Medical Language System (UMLS). METHODS: We exploited the fact that each Metathesaurus concept is assigned one or more semantic types from the UMLS Semantic Network and that the semantic types are arranged in a hierarchy. We compared the semantic types of each pair of parent and child concepts to determine if the types "explained" the Metathesaurus is-a relationships. We considered cases where the semantic type of the parent was neither the same as, nor an ancestor of, the semantic type of the child to be "unexplained." We applied this method to the January 2002 release of the UMLS and examined the unexplained cases we discovered to determine their causes. RESULTS: We found that 17022 (24.3%) of the parent-child is-a relationships in the UMLS Metathesaurus could not be explained based on the semantic types of the concepts. Causes for these discrepancies included cases where the parent or child was missing a semantic type, cases where the semantic type of the child was too general or the semantic type of the parent was too specific, cases where the parent-child relationship was incorrect, and cases where an ancestor-descendant relationship should be added to the UMLS Semantic network. In many cases, the specific cause of the discrepancy cannot be resolved without authoritative judgment by the UMLS developers. CONCLUSIONS: Our method successfully detects inconsistencies between the hierarchies of the UMLS Metathesaurus and Semantic Network. We believe that our method should be added to the set of tools that the UMLS developers use to maintain and audit the UMLS knowledge sources. PMID- 14759819 TI - The interaction of domain knowledge and linguistic structure in natural language processing: interpreting hypernymic propositions in biomedical text. AB - Interpretation of semantic propositions in free-text documents such as MEDLINE citations would provide valuable support for biomedical applications, and several approaches to semantic interpretation are being pursued in the biomedical informatics community. In this paper, we describe a methodology for interpreting linguistic structures that encode hypernymic propositions, in which a more specific concept is in a taxonomic relationship with a more general concept. In order to effectively process these constructions, we exploit underspecified syntactic analysis and structured domain knowledge from the Unified Medical Language System (UMLS). After introducing the syntactic processing on which our system depends, we focus on the UMLS knowledge that supports interpretation of hypernymic propositions. We first use semantic groups from the Semantic Network to ensure that the two concepts involved are compatible; hierarchical information in the Metathesaurus then determines which concept is more general and which more specific. A preliminary evaluation of a sample based on the semantic group Chemicals and Drugs provides 83% precision. An error analysis was conducted and potential solutions to the problems encountered are presented. The research discussed here serves as a paradigm for investigating the interaction between domain knowledge and linguistic structure in natural language processing, and could also make a contribution to research on automatic processing of discourse structure. Additional implications of the system we present include its integration in advanced semantic interpretation processors for biomedical text and its use for information extraction in specific domains. The approach has the potential to support a range of applications, including information retrieval and ontology engineering. PMID- 14759820 TI - A reference ontology for biomedical informatics: the Foundational Model of Anatomy. AB - The Foundational Model of Anatomy (FMA), initially developed as an enhancement of the anatomical content of UMLS, is a domain ontology of the concepts and relationships that pertain to the structural organization of the human body. It encompasses the material objects from the molecular to the macroscopic levels that constitute the body and associates with them non-material entities (spaces, surfaces, lines, and points) required for describing structural relationships. The disciplined modeling approach employed for the development of the FMA relies on a set of declared principles, high level schemes, Aristotelian definitions and a frame-based authoring environment. We propose the FMA as a reference ontology in biomedical informatics for correlating different views of anatomy, aligning existing and emerging ontologies in bioinformatics ontologies and providing a structure-based template for representing biological functions. PMID- 14759821 TI - OQAFMA Querying agent for the Foundational Model of Anatomy: a prototype for providing flexible and efficient access to large semantic networks. AB - The development of large semantic networks, such as the UMLS, which are intended to support a variety of applications, requires a flexible and efficient query interface for the extraction of information. Using one of the source vocabularies of UMLS as a test bed, we have developed such a prototype query interface. We first identify common classes of queries needed by applications that access these semantic networks. Next, we survey StruQL, an existing query language that we adopted, which supports all of these classes of queries. We then describe the OQAFMA Querying Agent for the Foundational Model of Anatomy (OQAFMA), which provides an efficient implementation of a subset of StruQL by pre-computing a variety of indices. We describe how OQAFMA leverages database optimization by converting StruQL queries to SQL. We evaluate the flexibility and efficiency of our implementation using English queries written by anatomists. This evaluation verifies that OQAFMA provides flexible, efficient access to one such large semantic network, the Foundational Model of Anatomy, and suggests that OQAFMA could be an efficient query interface to other large biomedical knowledge bases, such as the Unified Medical Language System. PMID- 14759822 TI - A physiological basis for biosphere-atmosphere interactions in the boreal forest: an overview. AB - Interdisciplinary field experiments for global change research are large, intensive efforts that study the controls on fluxes of carbon, water, trace gases, and energy between terrestrial ecosystems and the atmosphere at a range of spatial scales. Forest ecophysiology can make significant contributions to such efforts by measuring, interpreting, and modeling these fluxes for the individual components of forest ecosystems and then integrating the results into holistic ecosystem process models. The Boreal Ecosystem-Atmosphere Study (BOREAS) was undertaken because of the importance of the boreal forest biome to various global change issues. The study was conducted from 1993 to 1996 at sites in Saskatchewan and Manitoba, Canada. Results have shown that physiological processes of plants in the boreal forest can have large-scale consequences. For example, the composition of tree species strongly influences flux rates, with deciduous species having much higher carbon and water fluxes than coniferous species. Additionally, physiological limitations to transpiration in boreal conifers, even when soil water is abundant, reduces latent heat flux and increases sensible heat flux over large regions. This physiological control of transpiration can increase the depth of the atmospheric boundary layer on warm spring days to a level similar to that found in desert biomes. This special issue features 10 articles that address various aspects of the physiological basis of biosphere-atmosphere interactions in the boreal forest. The articles emphasize the environmental controls on water flux, carbon flux, and ecosystem productivity. PMID- 14759823 TI - Sap flow in trembling aspen: implications for stomatal responses to vapor pressure deficit. AB - We monitored sap flow and estimated diurnal changes in transpiration rates of two trembling aspen (Populus tremuloides Michx.) stands, located in the southern boreal forest and aspen parkland of Saskatchewan, Canada. In both stands during the peak growing season (June and July), sap flow during the day (0700-1700 h local time) increased linearly with vapor pressure deficit (VPD) from 0 to about 1 kPa, but then remained remarkably constant at VPD > 1 kPa (up to 4.8 kPa in the aspen parkland stand). The results imply an inverse relationship between stomatal conductance and VPD under well mixed conditions, for VPD > 1 kPa. We postulate that the stomata are operating to maintain leaf water potentials above a critical minimum value, which in turn places a maximum value on the rate of sap flow that can be sustained. PMID- 14759824 TI - Transpiration of a boreal pine forest measured by branch bag, sap flow and micrometeorological methods. AB - Three independent methods were used to evaluate transpiration of a boreal forest: the branch bag, sap flow and eddy covariance methods. The branch bag method encloses several thousand needles and gives a continuous record of branch transpiration. The sap flow method provides a continuous record of sap velocity and an estimate of tree transpiration. The eddy covariance method typically measures evaporation rates between a forest and the atmosphere. We deployed an extra eddy covariance system below the forest to estimate canopy transpiration by difference. The three systems detected small water vapor fluxes despite a plentiful supply of energy to drive evaporation. We also observed that transpiration rates were low even when the soil was well supplied with water. Low rates of transpiration were attributed to the canopy's low leaf area index and the marked reduction in stomatal conductance as vapor pressure deficits increased. Water vapor fluxes, derived from the sap flow method, lagged behind those derived by the branch bag method by 1 to 2 h. The sap flow method also suffered from sampling errors caused by the non-uniformity of flow across the sapwood and the spatial variability of sapwood cross section throughout the forest. Despite technical difficulties associated with hourly measurements, daily totals of transpiration agreed well with values derived from micrometeorological systems. PMID- 14759825 TI - Regulation of branch-level gas exchange of boreal trees: roles of shoot water potential and vapor pressure difference. AB - Effects of shoot water potential (Psi) and leaf-to-atmosphere vapor pressure difference (VPD) on gas exchange of jack pine (Pinus banksiana Lamb.), black spruce (Picea mariana (Mill.) B.S.P.), and aspen (Populus tremuloides Michx.) were investigated at the northern edge of the boreal forest in Manitoba, Canada. Laboratory measurements on cut branches showed that net photosynthesis (A(n)) and mesophyll conductance (g(m)) of jack pine and g(m) of black spruce did not respond to Psi until a threshold Psi was reached below which they decreased linearly. Photosynthesis of black spruce decreased slowly with decreasing Psi above the threshold and declined more rapidly thereafter. The threshold Psi was lower in black spruce than in jack pine. However, stomatal conductance (g(s)) of black spruce decreased continuously with decreasing Psi, whereas g(s) of jack pine showed a threshold response. Mesophyll limitations were primarily responsible for the decline in A(n) at low Psi for jack pine and black spruce in the middle of the growing season, but stomatal limitations became more important later in the season. Field measurements on in situ branches on warm sunny days showed that both conifer species maintained Psi above the corresponding threshold and there was no evidence of Psi limitation on A(n) of jack pine, black spruce or aspen. Vapor pressure difference was important in regulating gas exchange in all three species. An empirical model was used to quantify the g(s) response to VPD. When parameterized with laboratory data for the conifers, the model also fit the corresponding field data. When parameterized with field data, the model showed that stomata of aspen were the most sensitive of the three species to VPD, and stomata of black spruce were the least sensitive. For jack pine and aspen, stomata of foliage in the upper canopy were significantly more sensitive than stomata of foliage in the lower canopy. Vapor pressure difference had a greater impact on A(n) of aspen than on A(n) of the conifers as a result of aspen's greater stomatal sensitivity to VPD and greater slope of the relationship between A(n) and intercellular CO(2) concentration (C(i)). During the 1994 growing season, VPD averaged 1.0 kPa, corresponding to ratios of C(i) to ambient CO(2) of 0.77, 0.71 and 0.81 for jack pine, black spruce and aspen, respectively. We conclude that increases in VPD at the leaf surface in response to climate change should affect the absolute CO(2) and H(2)O fluxes per unit leaf area of the aspen component of a boreal forest landscape more than those of the conifer component. PMID- 14759826 TI - Automated measurements of CO(2) exchange at the moss surface of a black spruce forest. AB - We used an automated, multiplexing gas-exchange system to measure the net exchange of CO(2) at the surfaces of three shady feather moss and three exposed sphagnum moss sites in a black spruce forest during 35 days at the end of the 1995 growing season. Midday gross photosynthesis was 0.5 to 1.0 micro mol m(-2) s(-1) by feather moss and 0.5 to 2.5 micro mol m(-2) s(-1) by sphagnum moss. Photosynthesis by sphagnum moss was reduced by approximately 70% at 0 degrees C, and reached a maximum rate at 8 degrees C. Nighttime CO(2) efflux, the sum of soil and moss respiration was 1 to 2.5 micro mol m(-2) s(-1) above feather moss and 0.5 to 1.5 micro mol m(-2) s(-1) above sphagnum moss at moss temperatures of 0 to 15 degrees C. The higher rates of respiration at the feather moss sites probably reflected a greater belowground input of carbon from black spruce, and the lower rates of photosynthesis were probably associated with shading by the black spruce canopy. Photosynthesis by moss accounted for 10 to 50% of whole forest gross CO(2) uptake measured simultaneously by eddy covariance. Respiration at the moss surface was 50 to 90% of whole-forest respiration, with a decreasing fraction on warm nights apparently because of a disproportionate rise in aboveground respiration. PMID- 14759827 TI - Growth and maintenance respiration rates of aspen, black spruce and jack pine stems at northern and southern BOREAS sites. AB - We measured stem respiration rates during and after the 1994 growing season of three common boreal tree species at sites near the northern and southern boundaries of the closed-canopy boreal forest in central Canada. The growth respiration coefficient (r(g); carbon efflux per micro mole of carbon incorporated in structural matter) varied between 0.25 and 0.76, and was greatest for black spruce (Picea mariana (Mill.) B.S.P.), least for jack pine (Pinus banksiana Lamb.) and intermediate for trembling aspen (Populus tremuloides Michx.). There was a consistent trend for higher r(g) at northern sites than at southern sites. Maintenance respiration rates at 15 degrees C (r(m)) varied from 0.5 to 2.7 nmol C mol(-1) C(sapwood) s(-1). Values of r(m) were high at sapling stage jack pine sites and mature black spruce sites, and low at mature trembling aspen and mature jack pine sites. We found significant relationships between annual maintenance respiration and sapwood relative growth rate and sapwood volume per unit of stem surface area that explained much of the within-stand and between-stand variability. Because of the large differences in parameter values among sites, we conclude that the use of stand-specific respiratory parameters may improve model predictions of ecosystem process models over the use of generic parameter values. PMID- 14759828 TI - Variability in leaf-level CO(2) and water fluxes in Pinus banksiana and Picea mariana in Saskatchewan. AB - We measured seasonal and canopy-level gas exchange in two stands of jack pine (Pinus banksiana Lamb.) and one stand of black spruce (Picea mariana (Mill.) B.S.P.) on relatively clear days from late May until mid-September 1994. Field measurements were made with a portable infrared gas analyzer, and laboratory measurements included photosynthetic oxygen evolution and needle chemical composition. Seasonally averaged light-saturated assimilation rates in the field were 4.0 micro mol m(-2) s(-1) in jack pine and 2.7 micro mol m(-2) s(-1) in black spruce. Rates of assimilation and transpiration were highest in midsummer. The seasonal pattern was especially pronounced for black spruce, probably because cold soil temperatures limited early season gas exchange rates in this species. Among stands, instantaneous water-use efficiency was highest in a young jack pine stand early in the season and higher in the upper canopy foliage than in the lower canopy foliage at all sites at the end of the season. Needles of young jack pine exhibited higher photosynthetic capacity, dark respiration and needle N concentrations than needles of trees at the old site. In both species, slight acclimation to shading was manifested by reductions in photosynthetic capacity in the lower canopy foliage. In both species, first-year needles had greater photosynthetic capacity than older needles but in situ rates of CO(2) assimilation in the field showed little difference among needle age classes. In both species, there was a strong correlation between assimilation and stomatal conductance, indicating that assimilation was highly stomatal limited and that environmental factors that alter conductance (e.g., VPD) have a strong influence on CO(2) and water fluxes, especially after early season thawing concludes. PMID- 14759829 TI - Controls over monoterpene emissions from boreal forest conifers. AB - We investigated controls over the emission of monoterpenes from two species of boreal forest conifers, black spruce (Picea mariana Miller (B.S.P.)) and jack pine (Pinus banksiana Lamb). Monoterpenes are important in plants as carbon-based defensive compounds and in the atmosphere as photochemically reactive compounds that affect ozone and carbon monoxide concentrations. We examined ecological theories of plant allocation to defensive compounds in relation to emission rates of monoterpenes from the foliage of these two species. Monoterpene emission from plants is controlled by the vapor pressure of the monoterpenes within plant tissues, and vapor pressure is controlled by two parameters, air temperature and terpene concentration within the tissues. We measured the concentration of terpenes and nitrogen within foliage and the emission rate from foliage, and demonstrated that emission rate was linearly related to nitrogen concentration and exponentially related to air temperature. Current theories of plant allocation to carbon-based defenses predict an inverse relationship between foliar nitrogen and carbon-based defenses. We found that, under certain circumstances, these theories were sufficient to predict concentrations and emissions, but under other circumstances, the theories did not predict monoterpene concentrations or emissions. These results are discussed in the context of landscape/regional modeling of hydrocarbon emission from vegetation. PMID- 14759830 TI - Estimation of leaf area with an integrating sphere. AB - Relative absorptance of intact branches measured with an integrating sphere was compared to leaf area estimated by conventional methods (volume displacement and scanning area meter) for three conifer species: Picea mariana (Mill.) BSP, Pinus banksiana (Lamb.) and Pseudotsuga menziesii (Mirb.) Franco. A consistent relationship between relative absorptance and surface area emerged for the three species. The ability to predict leaf area from absorptance was further explored by measuring branches of Pseudotsuga menziesii grown in varying light and nutrient regimes. When a single equation was used to predict leaf area under all growth conditions, errors were as large as 40% primarily because of variation in leaf absorptivity, with the largest errors associated with extremely nutrient deficient foliage. When separate empirical equations were developed for each growth treatment, predicted leaf surface area agreed to within 5% of the area determined by the volume displacement method. Leaf surface area estimated from theoretical principles was also in good agreement with total surface area estimated independently by conventional methods. With proper accounting for needle absorptivity, which varied with growth conditions, leaf area estimates obtained by the integrating sphere method were of similar accuracy to those obtained by conventional methods, with the added advantage that the method allowed intact foliage to be sampled nondestructively in the field. Because the integrating sphere method preserves branch structure during measurement, it could provide a useful measure of needle area for photosynthetic or developmental studies requiring repeated sampling of the same branch. PMID- 14759831 TI - Root mass, net primary production and turnover in aspen, jack pine and black spruce forests in Saskatchewan and Manitoba, Canada. AB - Root biomass, net primary production and turnover were studied in aspen, jack pine and black spruce forests in two contrasting climates. The climate of the Southern Study Area (SSA) near Prince Albert, Saskatchewan is warmer and drier in the summer and milder in the winter than the Northern Study Area (NSA) near Thompson, Manitoba, Canada. Ingrowth soil cores and minirhizotrons were used to quantify fine root net primary production (NPPFR). Average daily fine root growth (m m(-2) day(-1)) was positively correlated with soil temperature at 10-cm depth (r(2) = 0.83-0.93) for all three species, with black spruce showing the strongest temperature effect. At both study areas, fine root biomass (measured from soil cores) and fine root length (measured from minirhizotrons) were less for jack pine than for the other two species. Except for the aspen stands, estimates of NPPFR from minirhizotrons were significantly greater than estimates from ingrowth cores. The core method underestimated NPPFR because it does not account for simultaneous fine root growth and mortality. Minirhizotron NPPFR estimates ranged from 59 g m(-2) year(-1) for aspen stands at SSA to 235 g m(-2) year(-1) for black spruce at NSA. The ratio of NPPFR to total detritus production (aboveground litterfall + NPPFR) was greater for evergreen forests than for deciduous forests, suggesting that carbon allocation patterns differ between boreal evergreen and deciduous forests. In all stands, NPPFR consistently exceeded annual fine root turnover and the differences were larger for stands in the NSA than for stands in the SSA, whereas the difference between study areas was only significant for black spruce. The imbalance between NPPFR and fine root turnover is sufficient to explain the net accumulation of carbon in boreal forest soils. PMID- 14759832 TI - Simulating forest productivity and surface-atmosphere carbon exchange in the BOREAS study region. AB - A process-based, general ecosystem model (BIOME-BGC) was used to simulate daily gross primary production, maintenance and heterotrophic respiration, net primary production and net ecosystem carbon exchange of boreal aspen, jack pine and black spruce stands. Model simulations of daily net carbon exchange of the ecosystem (NEE) explained 51.7% (SE = 1.32 g C m(-2) day(-1)) of the variance in daily NEE derived from stand eddy flux measurements of CO(2) during 1994. Differences between measured and simulated results were attributed to several factors including difficulties associated with measuring nighttime CO(2) fluxes and model assumptions of site homogeneity. However, comparisons between simulations and field data improved markedly at coarser time-scales. Model simulations explained 66.1% (SE = 0.97 g C m(-2) day(-1)) of the variance in measured NEE when 5-day means of daily results were compared. Annual simulations of aboveground net primary production ranged from 0.6-2.4 Mg C ha(-1) year(-1) and were concurrent with results derived from tree increment core measurements and allometric equations. Model simulations showed that all of the sites were net sinks (0.1-4.1 Mg C ha(-1) year(-1)) of atmospheric carbon for 1994. Older conifer stands showed narrow margins between uptake of carbon by net photosynthesis and carbon release through respiration. Younger stands were more productive than older stands, primarily because of lower maintenance respiration costs. However, all sites appeared to be less productive than temperate forests. Productivity simulations were strongly linked to stand morphology and site conditions. Old jack pine and aspen stands showed decreased productivity in response to simulated low soil water contents near the end of the 1994 growing season. Compared with the aspen stand, the jack pine stand appeared better adapted to conserve soil water through lower daily evapotranspiration losses but also exhibited a narrower margin between daily net photosynthesis and respiration. Stands subjected to water stress during the growing season may exist on the edge between being annual sources or sinks for atmospheric carbon. PMID- 14759833 TI - Effect of measurement CO(2) concentration on sugar maple root respiration. AB - Accurate estimates of root respiration are crucial to predicting belowground C cycling in forest ecosystems. Inhibition of respiration has been reported as a short-term response of plant tissue to elevated measurement [CO(2)]. We sought to determine if measurement [CO(2)] affected root respiration in samples from mature sugar maple (Acer saccharum Marsh.) forests and to assess possible errors associated with root respiration measurements made at [CO(2)]s lower than that typical of the soil atmosphere. Root respiration was measured as both CO(2) production and O(2) consumption on excised fine roots ( 20,000 micro l l(-1). Root respiration was significantly affected by the [CO(2)] at which measurements were made for both CO(2) production and O(2) consumption. Root respiration was most sensitive to [CO(2)] near and below normal soil concentrations (< 1500 micro l l(-1)). Respiration rates changed little at [CO(2)]s above 3000 micro l l(-1) and were essentially constant above 6000 micro l l(-1) CO(2). These findings call into question estimates of root respiration made at or near atmospheric [CO(2)], suggesting that they overestimate actual rates in the soil. Our results indicate that sugar maple root respiration at atmospheric [CO(2)] (350 micro l l(-1)) is about 139% of that at soil [CO(2)]. Although the causal mechanism remains unknown, the increase in root respiration at low measurement [CO(2)] is significant and should be accounted for when estimating or modeling root respiration. Until the direct effect of [CO(2)] on root respiration is fully understood, we recommend making measurements at a [CO(2)] representative of, or higher than, soil [CO(2)]. In all cases, the [CO(2)] at which measurements are made and the [CO(2)] typical of the soil atmosphere should be reported. PMID- 14759834 TI - Effects of dessication on post-planting stress in bare-root Corsican pine seedlings. AB - We examined the post-planting consequences of pre-planting exposure stress on two year-old, bare-root Corsican pine (Pinus nigra Arnold. ssp. laricio var. Corsicana) seedlings. Seedlings were lifted from a nursery and exposed to ambient conditions for periods of up to 192 h before being planted in minirhizotrons. Exposure decreased seedling water potential, CO(2) assimilation rate, leaf conductance and new root elongation, and increased mortality after planting. During exposure, needle total nonstructural carbohydrates (TNC) concentration (expressed on a dry mass basis) decreased by 0.31 mg g(dm) (-1) h(-1); however, needle and root TNC concentrations remained high (> 100 mg g(dm) (-1)) at planting, even in those treatments leading to severe seedling mortality. More than 90% of the seedlings with predawn water potentials lower than -1.3 MPa at planting did not elongate new roots and did not survive, whereas a similar percentage of seedlings with a predawn water potential above this value at planting elongated new roots and survived, suggesting that this value corresponds to a turgor threshold below which new root formation is inhibited. At planting, embolization of xylem conduits in roots and shoots was low for seedlings in all of the exposure treatments. PMID- 14759835 TI - Seasonal photosynthetic responses to light and temperature in white spruce (Picea glauca) seedlings planted under an aspen (Populus tremuloides) canopy and in the open. AB - Photosynthetic light and temperature response curves were measured seasonally in seedlings of white spruce (Picea glauca (Moench.) Voss) grown for two years in the understory of aspen (Populus tremuloides Michx.) or in the open in central Alberta. Light-saturated rate of net photosynthesis, the optimum temperature for net photosynthesis, transpiration rate, photochemical efficiency, and stomatal and mesophyll conductances increased from spring to summer and declined thereafter, whereas dark respiration rate and compensation and saturation points were highest in spring. Depression of photosynthetic parameters was greater in open-grown seedlings than in understory seedlings during the periods in spring and autumn when night frosts were common. Net photosynthetic rates were similar in understory and open-grown seedlings in summer, but they were significantly lower in open-grown seedlings in spring and autumn. Significantly lower transpiration rates and stomatal conductances in open-grown seedlings than in understory seedlings were also observed at 15 and 25 degrees C in the autumn. Shoot and needle growth were less in open-grown seedlings than in understory seedlings. In summer, when irradiances were low in the aspen understory, understory white spruce seedlings maintained a positive carbon balance by decreasing their compensation and saturation points and increasing their photochemical efficiency compared to spring and autumn. PMID- 14759836 TI - Influence of irradiance on water relations and carbon flux during rooting of Shorea leprosula leafy stem cuttings. AB - Single-node leafy stem cuttings of Shorea leprosula Miq. were subjected to a high, intermediate or low irradiance treatment for 16 weeks in an enclosed mist propagation system. Before rooting, maximum photosynthesis of the cuttings occurred at an irradiance of 400 micro mol m(-2) s(-1). Although none of the irradiance treatments affected the number of roots produced per cutting, the numbers of cuttings that formed roots were 50 and 30% in the high irradiance (diurnal range of 0-658 micro mol m(-2) s(-1)) and low irradiance (diurnal range of 0-98 micro mol m(-2) s(-1)) treatments, respectively, compared with 62% in the intermediate irradiance treatment (diurnal range of 0-360 micro mol m(-2) s(-1)). Low rooting frequency of cuttings in the high irradiance treatment was associated with water deficits (maximum leaf-to-air vapor pressure deficit (VPD) = 3.6 kPa), whereas cuttings in the low irradiance treatment had a low rooting frequency because they were below the light compensation point most of the time. In the intermediate irradiance treatment, cuttings withstood a daily maximum VPD of 1-2 kPa and recovered overnight from the previous day's deficit, as indicated by higher relative water content (RWC) and stomatal conductance (g(s)) in the morning than in the previous afternoon and evening. Higher RWC and g(s) of cuttings in all treatments on Days 14 and 21 compared with Day 8 probably indicated recovery from water deficit following severance and insertion of the cuttings in rooting medium. There were negative relationships between stem volume of cuttings and both number of cuttings that rooted and number of roots per cutting. PMID- 14759838 TI - Carbon isotopic composition, gas exchange, and growth of three populations of ponderosa pine differing in drought tolerance. AB - Effects of water supply on gas exchange, carbon isotopic composition, and relative growth rate were compared among seedlings from three populations of ponderosa pine (Pinus ponderosa Dougl. ex Laws.) grown in a controlled environment chamber. The three populations were chosen to represent high, moderate and low drought tolerance. There was no indication that drought tolerance was related to high water-use efficiency. Populations differed (P < 0.05) in relative growth rate (RGR), but did not differ (P > 0.10) in gas exchange variables or carbon isotope ratio (delta(13)C). Well-watered seedlings had significantly higher RGR, xylem pressure potential (Psi(xpp)), net photosynthesis (A), stomatal conductance to water vapor (g), and lower delta(13)C and instantaneous water-use efficiency than water-stressed seedlings. With decreasing Psi(xpp), A decreased linearly, whereas g decreased exponentially. Seedlings of the highly drought-tolerant population were more sensitive to water availability than seedlings from the other populations; they used water quickly when water was available, but closed their stomata in response to water stress. We conclude that, in ponderosa pine, the drought avoidance mechanism is more important for survival and growth in arid and semiarid environments than the efficient use of water. PMID- 14759837 TI - Effects of near-lethal heat stress on bud break, heat-shock proteins and ubiquitin in dormant poplar (Populus nigra Charkowiensis x P. nigra incrassata). AB - We assessed the effects of near-lethal heat stress on bud break, heat-shock proteins (HSPs) and ubiquitin in hybrid poplar (Populus nigra (L.) Charkowiensis x P. nigra (L.) incrassata). Shoots, with 10-15 buds each, were collected from September to March and exposed to temperatures between 20 and 60 degrees C for 2 h. Shoots were then placed in a greenhouse at 18-22 degrees C with supplemental light and cumulative bud break was recorded over a 4-week period. Samples of bud tissues were collected during and up to 96 h after heat treatment for protein analysis. De novo synthesis of proteins was monitored by exposing excised buds to [(35)S]-methionine for 3 h before, during, or after heat treatment. Heat treatments of 40-45 degrees C resulted in both a release from endodormancy and a decrease in thermal units needed for bud break during ecodormancy. The response to near-lethal heat stress was complex and was affected by intrinsic thermal sensitivity. Heat treatments were least effective during August and became progressively more effective as endodormancy progressed. In the later stages of ecodormancy, a heat treatment of 45 degrees C either inhibited bud break or killed the buds. Although temperatures of 42.5 to 45 degrees C inhibited incorporation of [(35)S]-methionine into proteins for at least 48 h, several HSPs were synthesized in response to temperatures of 40-45 degrees C. Immunoblots indicated that one of the heat-induced proteins was immunologically related to HSP70. Increases in free and conjugated forms of ubiquitin were also observed in response to heat treatment. Production of HSPs and ubiquitin, however, was not consistently associated with the heat treatments that induced the highest percentage of bud break. The roles of heat-induced protein degradation, HSPs, and ubiquitin in overcoming dormancy by near-lethal heat stress are discussed. PMID- 14759839 TI - Crop load and water stress effects on daily stem growth in peach (Prunus persica). AB - We investigated crop load and water stress effects on diurnal stem extension growth of field-grown peach (Prunus persica (L.) Batsch) trees. Neither the presence of fruit nor reduced irrigation significantly altered the timing of diurnal fluctuations in stem growth rate. Stems with subtending fruit had significantly reduced growth compared to stems with no subtending fruit. Crop load had no significant effect on relative stem extension rates and the majority of the reduction in absolute growth was the result of a smaller zone of elongation in fruit-bearing stems than in stems with no subtending fruit. Fruit removal did not increase growth rates within 24 h. When irrigation was reduced, the length of the stem elongation zone and total daily stem growth were significantly decreased relative to well-irrigated controls and the decreases were highly correlated with stem water potential. Compared with well-irrigated controls, relative stem extension rates of water-stressed trees were reduced at several times during the 24-h period, but the degree of reduction was not proportional to the difference in stem water potentials between the treatments. PMID- 14759840 TI - Carbon dioxide gas exchange of cembran pine (Pinus cembra) at the alpine timberline during winter. AB - Winter CO(2) gas exchange of the last three flushes of cembran pine (Pinus cembra L.) was studied under ambient conditions at the alpine timberline, an ecotone with strong seasonal changes in climate. During the coldest months of the year, December to March, gas exchange was almost completely suppressed and even the highest irradiances and temperatures did not cause a significant increase in net photosynthesis compared to spring and fall. In general, daily CO(2) balance was negative between December and March except during extended warm periods in late winter. However, because twig respiration was also reduced to a minimum during the December-March period, daily carbon losses were minimal. Total measured carbon loss during the winter months was small, equalling the photosynthetic production of one to two warm days in spring or summer when average air temperature was above 6 degrees C. PMID- 14759841 TI - Interdependence of pH, malate concentration, and calcium and magnesium concentrations in the xylem sap of beech roots. AB - The presence and concentration of mineral nutrients and organic acids were analyzed in root xylem sap of mature beech trees (Fagus sylvatica L.). An interdependence between malate concentration, pH, and calcium and magnesium concentrations was observed. Significant correlations were found between low pH values and high calcium and magnesium concentrations, low pH values and high malate concentration, and high malate concentration and high total calcium and magnesium concentrations. The observed correlations suggest that malic acid determines the chemical milieu of the xylem sap and tends to form complexes with hydrated or exchangeable adsorbed cations, thereby influencing the mobilization and translocation of calcium and magnesium in beech sapwood. PMID- 14759842 TI - Changes in rates of photosynthesis and respiration during needle development of loblolly pine. AB - Net photosynthetic rates of developing foliage and one-year-old foliage of loblolly pine (Pinus taeda L.) were measured under field conditions. In the subsequent year, net photosynthesis and dark respiration rates of current-year and one-year-old foliage were measured under controlled environmental conditions. Loblolly pine foliage grows slowly, reaching its final size 3.5 to 4 months after bud burst. Positive rates of net photosynthesis were recorded when the foliage was 13 and 18% of final length, in the controlled-environment and field study, respectively. However, because of high rates of dark respiration during the initial growth period, a positive diurnal carbon balance did not occur until foliage was about a third of final length (40 days after bud burst). Two months after bud burst, when foliage was about 55% of final length, its photosynthetic capacity exceeded that of one-year-old foliage. The highest rates of net photosynthesis were achieved when foliage was more than 90% fully expanded. PMID- 14759843 TI - Whole-plant hydraulic resistance and vulnerability segmentation in Acer saccharinum. AB - Hydraulic properties were studied in Acer saccharinum L., a riparian species that also grows well on a dry soil when transplanted. Hydraulic resistances were measured by two independent techniques: a new high-pressure flowmeter (HPFM) method and a conventional evaporative flux (EF) method. Vulnerability to cavitation was also investigated on petioles, stems and roots using a hydraulic conductivity technique. Vulnerability segmentation was found, i.e., roots, stems and petioles had different vulnerabilities to xylem dysfunction. Petioles were most vulnerable with 50% loss of hydraulic conductivity at -0.5 MPa, roots were least vulnerable (50% loss at -2.2 MPa) and stems were intermediate in vulnerability. The HPFM and the EF methods gave comparable results, except that the EF method gave a significantly higher value for resistance across petioles plus leaves. Native embolism was high enough to explain the discrepancy in resistance across petioles plus leaves between the HPFM and the EF methods, indicating that the HPFM estimates the minimum (potential) hydraulic resistance of plants. Whole-plant hydraulic resistance of A. saccharinum was low compared to resistances of other temperate species. The hydraulic characteristics of A. saccharinum were consistent with adaptation to its typical environment: low whole plant resistance assures high transpiration rates in the presence of sufficient water, and vulnerability segmentation provides the ability to survive during droughts through shedding of expendable organs. PMID- 14759844 TI - Hydraulic architecture and water relations of a flood-tolerant tropical tree, Annona glabra. AB - Hydraulic architecture parameters, water relation parameters and wood anatomy were studied in roots and shoots of the flood-tolerant tree Annona glabra L. on Barro Colorado Island, Panama. Hydraulic conductivity, leaf specific conductivity, and Huber value were similar to the corresponding values for tree species living in non-flooded habitats. The vulnerability of stems to loss of hydraulic conductivity resulting from embolism was low (50% loss of conductivity at -3.3 MPa). The lowest leaf water potential measured in the field was about 1.0 MPa, indicating that A. glabra has a large margin of safety from embolism, which may provide protection against rare drought events, or may be an adaptation to brackish mangrove habitats. Low absolute conductivity of roots was compensated for by an increase in the number of roots. More than two-thirds of whole-plant resistance to water flow was located in the roots. PMID- 14759845 TI - Acclimation of shade-developed leaves on saplings exposed to late-season canopy gaps. AB - We hypothesized that photoinhibition of shade-developed leaves of deciduous hardwood saplings would limit their ability to acclimate photosynthetically to increased irradiance, and we predicted that shade-tolerant sugar maple (Acer saccharum Marsh.) would be more susceptible to photoinhibition than intermediately shade-tolerant red oak (Quercus rubra L.). After four weeks in a canopy gap, photosynthetic rates of shade-developed leaves of both species had increased in response to the increase in irradiance, although final acclimation was more complete in red oak. However, photoinhibition occurred in both species, as indicated by short-term reductions in maximum rates of net photosynthesis and the quantum yield of oxygen evolution, and longer-term reductions in the efficiency of excitation energy capture by open photosystem II (PSII) reaction centers (dark-adapted F(v)/F(m)) and the quantum yield of PSII in the light (phi(PSII)). The magnitude and duration of this decrease were greater in sugar maple than in red oak, suggesting greater susceptibility to photoinhibition in sugar maple. Photoinhibition may have resulted from photodamage, but it may also have involved sustained rates of photoprotective energy dissipation (especially in red oak). Photosynthetic acclimation also appeared to be linked to an ability to increase leaf nitrogen content. Limited photosynthetic acclimation in shade developed sugar maple leaves may reflect a trade-off between shade-tolerance and rapid acclimation to a canopy gap. PMID- 14759846 TI - Nutrient availability alters belowground respiration of ozone-exposed ponderosa pine. AB - Exposure to ozone (O(3)) and changes in soil fertility influence both the metabolism of plant roots and their interaction with rhizosphere organisms. Because one indication of altered root metabolism is a change in belowground respiratory activity, we used specially designed measurement chambers to assess the effects of O(3) and nutrient availability on belowground respiratory activity of potted three-year-old ponderosa pine (Pinus ponderosa Dougl. ex Laws.). Seedlings were exposed to a factorial combination of three O(3) treatments and three fertilization treatments in open-top O(3) exposure chambers. Ozone exposure decreased and high nutrient supply increased total plant dry weight, but root/shoot ratios were not affected. In general, exposure to O(3) increased rates of belowground O(2) uptake and CO(2) release and the respiratory quotient (RQ, CO(2)/O(2)), although seasonal differences were detected. In October, following the second season of O(3) exposure, rates of belowground O(2) uptake and CO(2) release and RQ were increased in trees in the high-O(3) exposure treatment by 22, 73 and 32%, respectively, over values in control trees in charcoal-filtered air. Increasing nutrient supply resulted in decreasing rates of belowground O(2) uptake and CO(2) release but it had little effect on RQ. In the high-nutrient supply treatment, rates of belowground O(2) uptake and CO(2) release were decreased by 38 and 39%, respectively, compared with rates in the low-nutrient supply treatment. At the end of the second growing season, the high-nutrient supply treatment had decreased lateral root total nonstructural carbohydrates by 22% compared with the low-nutrient supply treatment. Nutrient availability altered the belowground respiratory response to O(3), such that the response to O(3) was greatest in the low-nutrient supply treatment. Significant O(3) effects on belowground respiratory activity were apparent before any reduction in total plant growth was found, suggesting that roots and rhizosphere organisms may be early indicators of physiological dysfunction in stressed seedlings. PMID- 14759847 TI - Effects of duration of a simulated winter thaw on dieback and xylem conductivity of Betula papyrifera. AB - Stems or roots + stems of potted, 2-year-old paper birch (Betula papyrifera L.) were subjected to simulated winter thaws of various durations in climate controlled chambers. The simulated thaws induced dieback of shoots of the treated plants. Although the stem thaw treatment did not significantly increase dieback, there were significant (P < 0.05) correlations between growing degree days above 4 degrees C and both shoot dieback and percent reduction in conductive xylem. All trees that received > 60 growing degree days (GDD) > 4 degrees C died back to some extent. Plants in the root + stem thaw treatment that received more than 60 GDD > 4 degrees C showed a significant (P < 0.05) increase in dieback and a significant (P < 0.05) loss of conducting xylem after a period of growth and recovery in the greenhouse, especially in the xylem of 1-year-old stems. Furthermore, higher correlations between GDD > 4 degrees C during a thaw and both the extent of dieback and the loss in conductive xylem were found in trees subjected to the root + stem thaw treatments than in trees exposed only to the stem thaw treatments (P < 0.05). The root + stem thaw treatments also resulted in highly significant relationships (P < 0.05-0.001) between loss in conductive xylem and dieback. The occurrence of dieback in response to winter thaws, and its close correlation with irreversible losses of xylem conductivity due to embolisms, coupled with an inability to refill the xylem because of root damage, support the view that these processes may be key factors in initiating birch decline. PMID- 14759848 TI - Effects of atmospheric CO(2) on longleaf pine: productivity and allocation as influenced by nitrogen and water. AB - Longleaf pine (Pinus palustris Mill.) seedlings were exposed to two concentrations of atmospheric CO(2) (365 or 720 micro mol mol(-1)) in combination with two N treatments (40 or 400 kg N ha(-1) year(-1)) and two irrigation treatments (target values of -0.5 or -1.5 MPa xylem pressure potential) in open top chambers from March 1993 through November 1994. Irrigation treatments were imposed after seedling establishment (i.e., 19 weeks after planting). Seedlings were harvested at 4, 8, 12, and 20 months. Elevated CO(2) increased biomass production only in the high-N treatment, and the relative growth enhancement was greater for the root system than for the shoot system. In water-stressed trees, elevated CO(2) increased root biomass only at the final harvest. Root:shoot ratios were usually increased by both the elevated CO(2) and low-N treatments. In the elevated CO(2) treatment, water-stressed trees had a higher root:shoot ratio than well-watered trees as a result of a drought-induced increase in the proportion of plant biomass in roots. Well-watered seedlings consistently grew larger than water-stressed seedlings only in the high-N treatment. We conclude that available soil N was the controlling resource for the growth response to elevated CO(2) in this study. Although some growth enhancement was observed in water-stressed trees in the elevated CO(2) treatment, this response was contingent on available soil N. PMID- 14759849 TI - Effects of cone-induction treatments on black spruce (Picea mariana) current-year needle development and gas exchange properties. AB - Both drought and root pruning (RP) increased the number of cones induced when black spruce (Picea mariana (Mill.) B.S.P.) grafts were injected with gibberellins A(4/7) (GA), but their effects on predawn shoot water potential and current-year needle development differed. Drought decreased predawn shoot water potential (Psi(pd)), but only during the period when irrigation was withheld, and it had no effect on the growth or gas exchange properties of current-year needles. Conversely, root pruning had little effect on Psi(pd), but it resulted in trees with smaller current-year needles that had lower nitrogen and chlorophyll concentrations and reduced rates of gas exchange up to the later stages of shoot elongation compared with needles of control trees. These findings are discussed in relation to potential effects on the development of induced cones in the following growth cycle. PMID- 14759850 TI - Seasonal courses of CO(2) exchange and carbon balance in fruits of Cinnamomum camphora. AB - Carbon dioxide exchange in fruits of Cinnamomum camphora Sieb. was followed over a growing season from July to December 1992. Dark respiration was exponentially related to temperature, with a Q(10) value near 2. Light dependence of photosynthetic CO(2) refixation, i.e., the ratio of gross photosynthesis to dark respiration, was approximated by a hyperbolic function. Seasonal variation in maximum CO(2) refixation capacity ranged between 52 and 174%, reaching a maximum in early August. Daily photosynthetic CO(2) refixation ranged between 17 and 51% over the growth period. We evaluated seasonal variation in translocation rate to the fruit on the basis of the seasonal rates of gross photosynthesis, dark respiration and increase in fruit dry weight, and used the results to develop a simple carbon flow model of fruit development. Seasonal changes in translocation rate paralleled those in fruit growth rate, with two peaks during the periods before and after September. Seed formation took place in the period between the two peaks. The relationship between fruit growth rate and translocation rate was approximated by a linear function. The carbon flow model estimated that, over the reproductive period, the amount of assimilate translocated to each fruit was 377.2 mg dry weight, of which 58.5% was accounted for by weight growth and 41.5% was consumed by net respiration. Carbon dioxide refixation accounted for 22.9% of the carbon balance of the fruit. PMID- 14759851 TI - Stomatal conductance, growth and root signaling in young oak seedlings subjected to partial soil drying. AB - Leaf conductance, water relations, growth, and abscisic acid (ABA) concentrations in xylem sap, root apices and leaves were assessed in oak seedlings (Quercus robur L.) grown with a root system divided between two compartments and subjected to one of four treatments: (a) well watered, WW; (b) half of root system exposed to soil drying and half kept well watered, WD; (c) whole root system exposed to drought, DD; and (d) half of root system severed, RE. Sharp decreases in plant stomatal conductance, leaf water potential, hydraulic conductance and leaf growth were observed during DD treatment. No significant differences in plant leaf water potential and stomatal conductance were detected between the WW and WD treatments. Nevertheless, the WD treatment resulted in inhibition of leaf expansion and stimulation of root elongation only in the well-watered compartment. Abscisic acid concentrations did not change in leaves, root tips, or xylem sap of WD- compared to WW-treated plants. Increased concentrations of ABA were observed in xylem sap from DD-treated plant roots, but the total flux of ABA to shoots was reduced compared to that in WW-treated plants, because of decreases in transpiration flux. Similar plant responses to the WD and RE treatments indicate that the responses observed in the WD-treated plants were probably not triggered by a positive signal originating from drying roots. PMID- 14759852 TI - Diurnal and seasonal changes in the impact of CO(2) enrichment on assimilation, stomatal conductance and growth in a long-term study of Mangifera indica in the wet-dry tropics of Australia. AB - We studied assimilation, stomatal conductance and growth of Mangifera indica L. saplings during long-term exposure to a CO(2)-enriched atmosphere in the seasonally wet-dry tropics of northern Australia. Grafted saplings of M. indica were planted in the ground in four air-conditioned, sunlit, plastic-covered chambers and exposed to CO(2) at the ambient or an elevated (700 micro mol mol( 1)) concentration for 28 months. Light-saturating assimilation (A(max)), stomatal conductance (g(s)), apparent quantum yield (phi), biomass and leaf area were measured periodically. After 28 months, the CO(2) treatments were changed in all four chambers from ambient to the elevated concentration or vice versa, and A(max) and g(s) were remeasured during a two-week exposure to the new regime. Throughout the 28-month period of exposure, A(max) and apparent quantum yield of leaves in the elevated CO(2) treatment were enhanced, whereas stomatal conductance and stomatal density of leaves were reduced. The relative impacts of atmospheric CO(2) enrichment on assimilation and stomatal conductance were significantly larger in the dry season than in the wet season. Total tree biomass was substantially increased in response to atmospheric CO(2) enrichment throughout the experimental period, but total canopy area did not differ between CO(2) treatments at either the first or the last harvest. During the two-week period following the change in CO(2) concentration, A(max) of plants grown in ambient air but measured in CO(2)-enriched air was significantly larger than that of trees grown and measured in CO(2)-enriched air. There was no difference in A(max) between trees grown and measured in ambient air compared to trees grown in CO(2)-enriched air but measured in ambient air. No evidence of down-regulation of assimilation in response to atmospheric CO(2) enrichment was observed when rates of assimilation were compared at a common intercellular CO(2) concentration. Reduced stomatal conductance in response to atmospheric CO(2) enrichment was attributed to a decline in both stomatal aperture and stomatal density. PMID- 14759853 TI - Histology of magnesium-deficient Norway spruce needles influenced by nitrogen source. AB - Effects of magnesium deficiency and variation in nitrate to ammonium ratio on needle histology and chlorophyll concentration were investigated in current-year and one-year-old needles of clonal Norway spruce trees (Picea abies (L.) Karst.). Six-year-old trees were grown for one year in sand culture with circulating nutrient solutions containing a sufficient (0.2 mM) or a limiting (0.04 mM) concentration of Mg. The nitrogen concentration was not varied (5 mM), but the NO(3) (-)/NH(4) (+)-ratio was adjusted to 0.76 in Mg-sufficient and to 1.86, 0.76 or 0.035 in Mg-limited plants. Visible symptoms of Mg deficiency occurred only in current-year needles, indicating adequate Mg nutrition before the experiment. Under conditions of Mg limitation, chlorophyll and Mg concentrations were lowest in needles of trees supplied with NH(4) (+) as the major nitrogen source and highest in needles of trees supplied with NO(3) (-) as the major nitrogen source. In current-year and one-year-old needles, starch accumulation induced by Mg deficiency was increased when NH(4) (+) was the major nitrogen source. The accumulation of tannin spherules in current-year needles, which occurred in response to Mg deficiency, also increased with decreasing NO(3) (-)/NH(4) (+) ratios. Deficient Mg supply caused premature aging in tissues of the vascular bundle, as indicated by modifications of the cambium and increased amounts of collapsed sieve cells. The number of collapsed sieve cells was slightly lower in needles grown in a NH(4) (+)-dominated nutrient regime than in needles grown in a NO(3) (-)-dominated nutrient regime. We conclude that was not directly toxic to Norway spruce trees at the applied concentrations. However, effects of Mg deficiency were considerably greater in an NH(4) (+)-dominated nutrient regime than in a NO(3) (-)-dominated nutrient regime. PMID- 14759854 TI - Root cold tolerance of black spruce seedlings: viability tests in relation to survival and regrowth. AB - Root systems of 6-month-old, cold-hardened, container-grown black spruce seedlings (Picea mariana (Mill.) B.S.P.) were exposed to 0, -5, -10, -15, -20, or -22.5 degrees C. Freezing-induced damage to fine roots, coarse roots and the whole root system was assessed by various viability tests including leakage of electrolytes, leakage of phenolic compounds, water loss, root and shoot water potentials, and live root dry mass. To assess the long-term effects of freezing induced root damage, seedling survival and regrowth were measured. Leakage of both electrolytes and phenolic compounds differed among fine roots, coarse roots, and whole root systems. In coarse roots and the whole root system, but not in fine roots, leakage of electrolytes, leakage of phenolic compounds, water loss, and root and shoot water potentials were correlated with percentage of live root dry mass which, in turn, was highly correlated with seedling survival and regrowth. Compared with live root dry mass, electrolyte and phenolic leakage, water loss, and root and shoot water potentials were less well correlated with seedling survival and regrowth. Among the viability tests, electrolyte leakage of the whole root system correlated most closely with seedling survival and regrowth. Under freezing conditions that destroyed less than 50% of each seedling's root system, about 70% of the seedlings survived and subsequent growth was little affected, whereas under freezing conditions that destroyed 70% of each seedling's root system, only about 30% of the seedlings survived and subsequent growth was reduced compared with that of undamaged plants. PMID- 14759855 TI - Effects of elevated CO(2) on chloroplast components, gas exchange and growth of oak and cherry. AB - Specific chloroplast proteins, gas exchange and dry matter production in oak (Quercus robur L.) seedlings and clonal cherry (Prunus avium L. x pseudocerasus Lind.) plants were measured during 19 months of growth in climate-controlled greenhouses at ambient (350 vpm) or elevated (700 vpm) CO(2). In both species, the elevated CO(2) treatment increased the PPFD saturated-rate of photosynthesis and dry matter production. After two months at elevated CO(2), Prunus plants showed significant increases in leaf (55%) and stem (61%) dry mass but not in root dry mass. However, this initial stimulation was not sustained: treatment differences in net assimilation rate (A) and plant dry mass were less after 10 months of growth than after 2 months of growth, suggesting acclimation of A to elevated CO(2) in Prunus. In contrast, after 10 months of growth at elevated CO(2), leaf dry mass of Quercus increased (130%) along with shoot (356%) and root (219%) dry mass, and A was also twice that of plants grown and measured at ambient CO(2). The amounts of Rubisco and the thylakoid-bound protein cytochrome f were higher in Quercus plants grown for 19 months in elevated CO(2) than in control plants, whereas in Prunus there was less Rubisco in plants grown for 19 months in elevated CO(2) than in control plants. Exposure to elevated CO(2) for 10 months resulted in increased mean leaf area in both species and increased abaxial stomatal density in Quercus. There was no change in leaf epidermal cell size in either species in response to the elevated CO(2) treatment. The lack of acclimation of photosynthesis in oak grown at elevated CO(2) is discussed in relation to the production and allocation of dry matter. We propose that differences in carbohydrate utilization underlie the differing long-term CO(2) responses of the two species. PMID- 14759856 TI - Effects of water stress cycles on turgor maintenance processes in pear leaves (Pyrus communis). AB - Effects of water deficits on leaf turgor maintenance processes were analyzed for pear trees (Pyrus communis L. cv. "Barlett") grown in 120-liter containers. Four irrigation treatments were applied: a well-watered control treatment, a spring water stress cycle (Sp), a summer water stress cycle (Su), and a spring plus summer water stress cycle (Sp + Su). For the Sp treatment, water application was progressively reduced from 100 to 20% of the control dose over a period of 27 days in spring. For the Su treatment, water application was progressively reduced over 23 days in summer, from 100 to 20% of the control dose. The Sp + Su treatment comprised both the spring and summer drought stress cycles. Pressure volume (P-V) curves were constructed and stomatal conductances were determined for pear leaves from each treatment during the spring and summer stress cycles. Leaf water potential (Psi(pi) (0)) and relative water content (R(0)) at the turgor loss point of control leaves tended to decrease from spring to summer. Changes in leaf osmotic water potential at full turgor (Psi(pi) (100)) and in symplast water fraction (R(s)) did not explain the seasonal decrease in Psi(pi) (0). The water stress treatments had no effect on Psi(pi) (100), but R(s) was reduced by the water stress treatments, particularly during the summer stress cycle of the Su and Sp + Su treatments. The decrease in R(s) was correlated with an increase in the slope of the linear region of the P-V curve. Such a coupled adjustment would lead to increased water uptake capacity of water-stressed trees only under non-turgor conditions. Furthermore, pear leaves did not actively accumulate solutes. We conclude, therefore, that changes in leaf tissue water relations as a result of leaf acclimation to water stress are unlikely to facilitate maintenance of fruit productivity under drought. PMID- 14759857 TI - Role of flavanols in yellowing beech trees of the Black Forest. AB - Beech leaves were sampled at the end of a prolonged hot dry period at a tree decline site in the Black Forest, Germany to investigate the potential role of flavanols in defense mechanisms against environmental stress. Green and yellowing leaves were harvested from the uppermost canopy of trees that were more than 200 years old and 30 m high. Yellowing leaves had a 7.4-fold higher concentration of total flavanols than green leaves. Green leaves contained flavanol inclusions, but during yellowing the inclusions disintegrated and the cells became filled with flavanols. Abscisic acid (ABA) stimulated the release of flavanols from intravacuolar inclusions of leaf petioles and flower pedicels. In addition, ABA caused flavanols to leach from the trichomes of beech galls. The antioxidative potential of leaf extracts, as estimated by indoleacetic acid (IAA) oxidation, was significantly higher in yellowing leaves than in green leaves. In vitro experiments revealed that (+)-catechin promoted growth of beech tissue. PMID- 14759858 TI - Influence of rewatering and time of sampling on solute accumulation of two Populus deltoides clones. AB - We investigated effects of rehydration and time of sampling on solute accumulation in plants of two greenhouse-grown Populus deltoides Bartr. clones (Ohio Red and Platte) subjected to multiple cycles of water stress. Osmotic potential of leaves at full turgor was measured at predawn on well-watered (control) and water-stressed (conditioned) plants that had been rewatered the previous afternoon. Water-soluble organic solutes (carbohydrates, phenolic glucosides and organic acids) were determined at both predawn and midday, before and after rewatering. Conditioning resulted in solute accumulation; however, rewatering of conditioned plants decreased the predawn concentration of glucose by 19-35% and the total solute concentration by 14-15%, relative to values before rewatering. There was a 52% increase in salicin concentration in response to rewatering in conditioned plants of the Platte clone. In conditioned plants of both clones, the concentration of glucose was generally lower at midday than at predawn (16-47%), whereas the concentration of sucrose was higher at midday than at predawn (46-133%). Time of sampling was an important factor in determining whether conditioning resulted in accumulation of glucose and fructose. Compared with control plants, there was a significant accumulation of glucose and fructose at predawn and either no accumulation or a significant reduction of these solutes at midday both before and after rewatering of conditioned plants. Sampling time also affected the amount of solute that accumulated in response to conditioning. PMID- 14759859 TI - Elevated atmospheric CO(2) concentration changes ectomycorrhizal morphotype assemblages in Betula papyrifera. AB - Ectomycorrhizae are extremely diverse, with different species of fungi having very different physiologies and morphologies that, in turn, confer a range of different benefits to the host plant. To test the hypothesis that elevated CO(2) leads to changes in the assemblage of ectomycorrhizae associated with trees, we examined the number and frequency of ectomycorrhizal morphotypes colonizing roots of Betula papyrifera Marsh. saplings grown at an ambient or elevated (700 ppm) atmospheric CO(2) concentration for 24 weeks. Elevated CO(2) resulted in significant changes in the composition of the ectomycorrhizal assemblage toward morphotypes with a higher incidence of emanating hyphae and rhizomorphs. We conclude that B. papyrifera saplings will be able to support a more costly mycorrhization in future elevated-CO(2) atmospheres. PMID- 14759860 TI - Time and method of floral initiation and effect of paclobutrazol on flower and fruit development in Shorea stenoptera (Dipterocarpaceae). AB - Small Shorea stenoptera Burck. (Dipterocarpaceae) trees of reproductive age growing in an arboretum in west Java were studied to determine the pattern of vegetative shoot development, the time and method of floral initiation and the effect of paclobutrazol on floral enhancement. Vegetative buds were enclosed by two stipules between which was a leaf primordium, a small axillary vegetative bud and another pair of stipules. This sequence was reiterated five to seven times before the vegetative apex was visible. At the time of floral initiation, axillary buds developed into floral spikes and compound inflorescences formed at the end of drooping branches. A compound inflorescence might bear many floral spikes and each floral spike bore many flowers. The compound inflorescence was a modification of the reiterative developmental pattern observed in vegetative shoots. The time of floral initiation began in late June or early July and continued until about November. Floral enhancement using paclobutrazol as a soil drench was attempted in mid-July, but this was later found to be after the onset of floral initiation, and the treatment failed to enhance flowering; however, it appeared to enhance the rate of floral and fruit development. The similarity in vegetative bud development among dipterocarp genera suggests that the time of floral initiation may be easily determined in many species based on simple dissection techniques. PMID- 14759861 TI - Effect of mineral nutrition content on oxygen exchange and chlorophyll a fluorescence in needles of Norway spruce. AB - Photosynthetic O(2) evolution at high irradiances (approximately 600-1000 micro mol m(-2) s(-1)) and O(2) uptake in darkness were measured in needles of control, irrigated and irrigated-fertilized trees of Norway spruce (Picea abies (L.) Karst.). Measurements were made at 20 degrees C and at high CO(2) concentrations. The results suggest that, at given times of the year, a major part of the variation in gross photosynthesis of current-year and one-year-old needles across treatments is associated with differences in needle N content. Furthermore, the rate of O(2) uptake measured after 5 or 10 min in darkness was positively correlated with both the preceding rate of gross O(2) evolution and the N content in fully expanded current-year needles. Measurements of chlorophyll a fluorescence, taken simultaneously with measurements of O(2) evolution in current year sun needles, showed that Stern-Volmer quenching of minimum fluorescence and the ratio of variable to maximum fluorescence in the dark- and light-adapted state were strongly correlated with the gross rate of O(2) evolution. This suggests that the increased rate of gross photosynthesis in needles of irrigated fertilized trees was associated with adjustments in the thermal energy dissipation within photosystem II. PMID- 14759862 TI - Photosynthetic responses of Scots pine to elevated CO(2) and nitrogen supply: results of a branch-in-bag experiment. AB - Naturally seeded Scots pine (Pinus sylvestris L.) trees, age 25-30 years, were subjected to two soil-nitrogen-supply regimes and to elevated atmospheric CO(2) concentrations by the branch-in-bag method from April 15 to September 15 for two or three years. Gas exchange in detached shoots was measured in a diffuse radiation field. Seven parameters associated with photosynthetic performance and two describing stomatal conductance were determined to assess the effects of treatments on photosynthetic components. An elevated concentration of CO(2) did not lead to a significant downward regulation in maximum carboxylation rate (V(cmax)) or maximum electron transport rate (J(max)), but it significantly decreased light-saturated stomatal conductance (g(sat)) and increased minimum stomatal conductance (g(min)). Light-saturated rates of CO(2) assimilation were higher (24-31%) in shoots grown and measured at elevated CO(2) concentration than in shoots grown and measured at ambient CO(2) concentration, regardless of treatment time or nitrogen-supply regime. High soil-nitrogen supply significantly increased photosynthetic capacity, corresponding to significant increases in V(cmax) and J(max). However, the combined elevated CO(2) + high nitrogen-supply treatment did not enhance the photosynthetic response above that observed in the elevated CO(2) treatment alone. PMID- 14759863 TI - Geographical variation in water relations, hydraulic architecture and terpene composition of Aleppo pine seedlings from Italian provinces. AB - Ecotypic variations in leaf conductance, soil-to-leaf hydraulic conductance, components of tissue water potential, hydraulic architecture parameters and xylem embolism were examined in greenhouse-grown two-year-old Aleppo pine (Pinus halepensis Mill.) seedlings from six origins representing the geographic range of the species in Italy. Cortical resin composition of the seedlings was also determined. Measurements were made on well-watered seedlings and on seedlings subjected to recurring severe drought. Drought-stressed seedlings had lower mean leaf conductances, transpiration rates and soil-to-leaf hydraulic conductances than well-watered seedlings. They also exhibited more negative osmotic potentials, higher relative water deficit at incipient plasmolysis, but a similar maximum modulus of elasticity. Drought-stressed seedlings showed a higher degree of xylem embolism, a lower Huber value, lower leaf specific conductivity and lower specific conductivity than well-watered seedlings. Drought-stressed seedlings of provenances from more xeric habitats (Tremiti, Porto Pino and Mottola) had greater leaf conductances, transpiration rates and soil-to-leaf hydraulic conductances than drought-stressed seedlings of provenances from more mesic habitats (Imperia, Otricoli and Vico del Gargano). They also showed higher osmotic adjustment and a lower degree of xylem embolism. Among provenances, there were no significant differences in hydraulic architecture parameters in response to the drought treatment; however, Tremiti and Porto Pino seedlings displayed smaller drought-induced reductions in specific conductivity and leaf specific conductivity, respectively, than seedlings from other provenances. These differences suggest that seedlings from xeric provenances, especially Tremiti, have greater resistance to desiccation than seedlings from mesic provenances. No clear association was found between terpene variability and the other traits investigated, although terpene composition was related to the geographical distribution of the provenances. We conclude that the drought-tolerance responses of Tremiti make it a more suitable provenance than the others for establishment on sites prone to severe soil water deficits. PMID- 14759864 TI - Branch growth and biomass allocation in Abies amabilis saplings in contrasting light environments. AB - Aboveground biomass allocation, and height and branch growth were studied in saplings of the shade-tolerant conifer, Abies amabilis Dougl. ex Forbes growing in large openings and in the understory of an old-growth forest in western Oregon. The presence of annual overwintering budscale scars was used to infer extension growth histories; annual growth rings in branches and stems were used in combination with extension histories to compute partitioning of new biomass among leaves, branches and stems. Saplings growing in large gaps had conical crowns, whereas understory saplings had umbrella shaped crowns as a result of much greater rates of branch extension than stem extension. Understory saplings grew slowly in height because of low rates of biomass production and low allocation of biomass to stem extension. About 40% of new biomass was allocated to foliage in both groups, but understory saplings allocated more of the remaining growth increment to branches and less to stem than did saplings growing in large gaps. These results differ from the patterns observed in shade-tolerant saplings of tropical forests, where allocation to foliage increases with shading and branch allocation is much lower than observed here. This difference in allocation may reflect mechanical constraints imposed by snow loads on the evergreen A. amabilis crowns, particularly on flat-crowned understory saplings. PMID- 14759865 TI - A simple model of light and water use evaluated for Pinus radiata. AB - An existing model of light and water use by crops (RESCAP) was adapted and evaluated for trees. In the model, growth on any given day is determined either by the amount of intercepted radiation (by means of the light utilization coefficient, epsilon) or by the maximum rate of water extraction by roots (a function of root biomass and soil water content). In either case, transpiration and growth are related by the water-use efficiency (q), which is inversely proportional to the daily mean saturation vapor pressure deficit (D). The model was applied to two Pinus radiata (D. Don) stands (control (C) and fertilized (F)) growing near Canberra, Australia, using data collected during the Biology of Forest Growth experiment (1983-1988). For both stands, predicted and measured soil water contents were in close agreement (r(2) > 0.9) over a 4-year period involving several wet-dry cycles. The parameter combination epsilon/qD was estimated to be 0.28 and 0.26 kg H(2)O (MJ total)(-1) kPa(-1) for the C and F stands, respectively. Because of the close physiological link between water use and CO(2) uptake, the results suggest that tree growth may be realistically simulated by simple models based on conservative values for epsilon and qD. PMID- 14759866 TI - Growth and water relations of Eucalyptus marginata (jarrah) stands in response to thinning and fertilization. AB - We studied the effects of five thinning treatments (T1 = 5.5, T2 = 11, T3 = 16.5, T4 = 22.5 and T5 = 28.5 m(2) ha(-1) basal area under bark) x two fertilizer treatments (F0 = unfertilized and F1 = fertilized with 400 kg ha(-1) N plus 229 kg ha(-1) P) on growth and water relations of pole-sized Eucalyptus marginata J. Donn ex Sm. trees growing in southwestern Australia. Thinning reduced leaf area index (LAI) from 2.1 in the T4 and T5 treatments to 0.8 in the T1F0 treatment. Fertilizer had no effect on LAI in the T2, T4 or T5 treatments, but increased LAI by 45 and 20% in the T1 and T3 treatments, respectively. Thinning plus fertilizing increased diameter growth most in the fastest growing trees, from 0.4 cm year(-1) for trees in the T5F0 and T5F1 treatments to 0.7 and 1.2 cm year(-1) for trees in the T1F0 and T1F1 treatments, respectively. In both fertilizer treatments, stand basal area and volume growth increased with increasing stand density up to 15 m(2) ha(-1), and thereafter declined with increasing stand density, such that the growth rate of trees in the T5 treatment was only half of that at a stand density of 15 m(2) ha(-1). In response to fertilizer, growth rates of the slowest and fastest-growing trees increased from 0.35 and 3.5 m(2) ha(-1) year(-1) (F0) to 0.56 and 5.4 m(3) ha(-1) year(-1) (F1), respectively. Stand growth efficiency (growth per unit LAI) increased in response to thinning, and fertilizer increased stand growth efficiency at all stand densities. Throughout the dry season, T5 trees had lower predawn shoot water potentials (Psi(pd)) (minimum of -1.5 MPa) than T1 or T2 trees (minimum of -0.7 MPa). Fertilizer decreased Psi(pd) in T5 trees (by -0.9 and -1.5 MPa, respectively, in F0 and F1), but not in T1 or T2 trees. Stand growth rate was closely related to cumulative midday water stress (CMWS) over the dry season, and volume growth rate declined sharply from 6 m(3) ha(-1) year(-1) at a CMWS of 130 MPa days, to zero at a CMWS of 220 MPa days. Application of fertilizer to thinned stands increased LAI, stand growth efficiency and stand growth. In unthinned stands, fertilizer increased stand growth efficiency and stand growth; however, it also increased tree water stress, which limited the fertilizer-induced increases in LAI and growth. We attribute the increase in tree and stand growth in response to application of fertilizer to increased photosynthetic rates, increased allocation to stem wood, and in thinned stands also to higher LAIs. PMID- 14759867 TI - Xylem cavitation in roots and stems of Douglas-fir and white fir. AB - Roots of hardwoods have been shown to be more vulnerable to xylem cavitation than stems. This study examined whether this pattern is also observed in a conifer species. Vulnerability to cavitation was determined from the pressure required to inject air into the vascular system of hydrated roots and stems, and reduce hydraulic conductance of the xylem. According to the air-seeding hypothesis for the cavitation mechanism, these air pressures predict the negative xylem pressure causing cavitation in dehydrating stems. This was evaluated for stems of Douglas fir (Pseudotsuga menziesii (Mirb.) Franco) and white fir (Abies concolor (Gord. & Glend.) Lindl.). The air-injection method was applied to roots and stems of different sizes and positions in Douglas-fir trees. Roots, especially smaller roots with a xylem diameter < 5 mm, were more vulnerable to cavitation than stems. Mean cavitation pressure for smaller roots was -2.09 +/- 0.42 versus -3.80 +/- 0.19 MPa for larger roots (> 8 mm diameter). Within the shoot system, smaller stems (< 5 mm diameter) were most vulnerable to cavitation, having a mean cavitation pressure of -4.23 +/- 0.565 versus -5.27 +/- 0.513 MPa for large stems (> 8 mm diameter). There was no correlation between tracheid diameter and mean cavitation pressure within root or stem systems, despite larger tracheid diameters in roots (23.3 +/- 3.9 micro m) than in stems (9.2 +/- 1.6 micro m). Smaller safety margins from cavitation in roots may be beneficial in limiting water use during mild drought, and in protecting the stem from low xylem pressures during extreme drought. PMID- 14759868 TI - Citrus response to salinity: growth and nutrient uptake. AB - To determine the effects of salinity on relative growth rate (RGR), net assimilation rate on a leaf weight basis (NAR(w)), leaf weight ratio (LWR), and nutrient uptake and utilization of citrus, we grew four citrus rootstocks (sour orange, Cleopatra mandarin, Carrizo citrange and Citrus macrophylla) in nutrient solutions containing 0, 10, 20, 40 or 80 mM NaCl for 20, 40 or 60 days. For each element analyzed, specific absorption rate (SAR) and specific utilization rate on a leaf basis (SUR(L)) were calculated for the period between Days 40 and 60. Relative growth rate decreased with time for all treatments and rootstocks. Salt treatment significantly reduced both RGR and NAR(w), whereas LWR showed no definite trend. In all rootstocks, NAR(w), but not LWR, was significantly correlated with RGR, indicating that NAR(w) was an important factor underlying the salinity-induced differences in RGR among the citrus rootstocks. At Day 60, salinity had a significant effect on leaf concentrations of Cl, Na, K, Ca, Mg, P, Fe, Mn and Zn and on the SAR and SUR(L) of most elements. In general, RGR was correlated with SAR and SUR(L). Therefore, in addition to osmotic effects and the inhibitory effects of high concentrations of Cl(-) and Na(+), an imbalance of essential nutrients may also contribute to the reduction in plant growth under saline conditions. PMID- 14759869 TI - Boxelder water sources and physiology at perennial and ephemeral stream sites in Arizona. AB - To assess the influence of stream water on leaf gas exchange and water potential in different sized boxelder trees (Acer negundo L.), we compared these characteristics in trees growing beside a perennial stream and a nearby ephemeral stream in a montane-riparian forest in northern Arizona. Patterns of tree water use were quantified by stable isotope analysis (delta(18)O). Physiological characteristics were similar for large and small trees. Similarity between sites in predawn and daytime water potentials and xylem delta(18)O indicated that stream water was not a physiologically important water source. Seasonal and site variations in light-saturated net photosynthetic rate were significantly related to leaf-to-air vapor pressure deficit (r = -0.691) and foliar nitrogen concentration (r = 0.388). Although deep water was the dominant water source, surface soil water was utilized following precipitation, especially by small trees. We conclude that net carbon gain and severity of water stress are only weakly coupled to stream water availability. PMID- 14759870 TI - Evidence for red:far red signaling and photomorphogenic growth response in Douglas-fir (Pseudotsuga menziesii) seedlings. AB - In a greenhouse experiment, potted coastal Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) seedlings were grown in miniature "Nelder" (Nelder 1962) plots where growing space varied from 265 to 2555 cm(2) per plant. After thirty weeks, mean plant height, crown biomass and branch number increased significantly (P = 0.0141) with decreasing growing space (increasing plant density). Differences in height growth became apparent about six weeks after sowing. Furthermore, horizontally reflected radiation measured within the Nelder plots showed a decrease in red:far-red ratio (R:FR) from 1.2 at the lowest density to 0.71 at the highest. Plant height was strongly inversely correlated with estimated phytochrome photoequilibrium values (r(2) = 0.893). Field measurements made in a three-year-old variable density plantation also showed a decrease in R:FR with increasing planting density from 300 to 3,000 trees ha(-1). These results support the hypothesis that young Douglas-fir seedlings are able to detect, through the phytochrome system, the presence of nearby seedlings owing to the depletion of R relative to FR in the spectra reflected by the foliage of the adjacent plants. They then adjust their growth allometry in a way that reduces the possibility of being over topped by these future competitors. PMID- 14759871 TI - Leaf nutrition and photosynthetic performance of sugar maple (Acer saccharum) in stands with contrasting health conditions. AB - Leaf nutrition and photosynthetic performance of sugar maple (Acer saccharum Marsh.) were compared between two sugar maple stands in northwestern Vermont with contrasting health conditions as indicated by annual basal area growth, degree of crown dieback, and foliar appearance. Observations made during the diurnal cycle of both stands showed no apparent leaf water stress. In both stands, leaves had similar concentrations of major non-structural carbohydrates (starch and sucrose). Over two consecutive growing seasons (1991 and 1992), we consistently observed lower leaf Ca and Mg concentrations in the declining stand than in the healthy stand. Compared with the healthy stand, lower leaf chlorophyll concentrations and apparent leaf chlorosis were observed in the declining stand, and some trees had very low foliar Ca and Mg concentrations (0.31 +/- 0.03% and 0.09 +/- 0.01%, respectively). Trees in the declining stand had lower light saturated net photosynthetic rates on a dry mass basis at both ambient CO(2) (P(n,amb)) and saturating CO(2) (P(n,sat)) than trees in the healthy stand. There were significant linear correlations between P(n,amb) and leaf mass per unit area (LMA) and between P(n,sat) per unit leaf area and LMA. There were also linear correlations between both P(n,amb) and P(n,sat) and leaf N when expressed on an area basis in both stands, indicating that variation in LMA may have been largely responsible for the observed photosynthesis-nitrogen relationship. The values of P(n,amb) and P(n,sat) were not significantly correlated with leaf N on a mass basis but were weakly correlated with leaf Ca and Mg on a mass basis. We conclude that low leaf Ca or Mg concentrations may limit leaf CO(2) assimilation and tree carbohydrate status in the declining stand. PMID- 14759872 TI - Root carbohydrate reserves, mineral nutrient concentrations and biomass in a healthy and a declining sugar maple (Acer saccharum) stand. AB - Soil and root characteristics were contrasted between a "declining" and a "healthy" sugar maple (Acer saccharum Marsh.) stand in Vermont, USA. The declining stand had lower basal area increment and more crown dieback than the healthy stand. Soil pH and base cation content were lower and soil water content was higher at the site of the declining stand than at the site of the healthy stand, whereas soil temperature did not differ significantly between the sites. In live fine roots, concentrations of K and Ca were marginally (P < 0.07) lower in the declining than in the healthy stand, whereas concentrations of N, P, Mg, and Al were not significantly different (P = 0.13 to 0.87) between stands. Starch and soluble sugar concentrations of fine and coarse roots did not differ significantly between stands, indicating that crown dieback did not affect carbohydrate supply to the roots in the declining stand. Throughout the growing season, the standing live and dead root biomass were significantly higher in the declining stand than in the healthy stand, indicating that more carbon was allocated to roots and that root turnover was higher in the declining stand than in the healthy stand. PMID- 14759873 TI - Seasonal air and soil temperature effects on photosynthesis in red spruce (Picea rubens) saplings. AB - Net photosynthesis and stomatal conductance were measured in ten red spruce (Picea rubens Sarg.) saplings, growing near Ithaca, New York, throughout the early spring and late-fall growing periods. Gas exchange and daily minimum and maximum soil and air temperatures were also measured. Linear regression analysis showed that rates of net photosynthesis were positively correlated with both minimum daily soil and air temperatures but that minimum soil temperature was a better predictor of net photosynthesis. Moreover, net photosynthesis was more sensitive to changes in soil temperature than to changes in air temperature, and photosynthesis was approximately twice as sensitive to temperature changes during the fall than during the spring. PMID- 14759874 TI - Water use by Eucalyptus tereticornis stands of differing density in southern India. AB - We studied water use by Eucalyptus tereticornis Sm. in two plantations, differing in tree density (1800 stems ha(-1) at Site I and 1090 stems ha(-1) at Site II), in different years. At both sites, stomatal conductance, predawn and midday water potentials and microclimate were measured and used to estimate hourly transpiration by the Penman-Monteith equation. Growth in girth was also measured. Stomatal conductance was closely correlated with atmospheric vapor pressure deficit (D); however, stomata did not close completely even at high D ( approximately 5.0 kPa). Midday leaf water potentials did not fall below -2.0 MPa during any part of the year at either site. Predawn leaf water potentials were greater than -0.25 MPa during the postmonsoon period, but declined to -0.7 MPa at Site I during the premonsoon period. Transpiration estimates ranged from 0.6 to 1.2 mm h(-1) at Site I and from 0.2 to 0.6 mm h(-1) at Site II. The extrapolated transpiration values for the rain-free days of the year were 1563 mm and 853 mm for Sites I and II, respectively. Growth in girth was negligible during the premonsoon period. Photosynthesis was not affected by the minor water stress that developed during the premonsoon period. PMID- 14759875 TI - Structural variation in current-year shoots of broad-leaved evergreen tree saplings under forest canopies in warm temperate Japan. AB - Stem length and leaf area of current-year shoots were measured in saplings of eight broad-leaved evergreen tree species growing under a forest canopy. Stem length varied over a range of one to two orders of magnitude within each species. In all species, both the number of leaves and the mean stem length between successive leaves were greater in longer shoots. Mean leaf size and stem length were not correlated in six of eight species, and only weakly positively correlated in the other two species. Thus, total leaf area per stem increased with stem length, but not in direct proportion: leaf area per stem length was smaller in shoots with long stems and larger in shoots with short stems. I conclude that the within-species variation in the leaf-stem balance of current year shoots is related to variation in shoot functional roles, as has been observed for long and short shoots in many deciduous tree species: shoots with long stems are extension oriented and contribute to the framework of the crown, whereas shoots with short stems serve mainly for leaf display. Among species, large differences were found in the leaf area per stem length ratio. In the species with larger leaf area per stem length ratios, leaves had narrower blades or longer petioles, or both, resulting in a reduction of mutual shading among the leaves on the shoot. PMID- 14759876 TI - Crown architecture of Abies balsamea from four canopy positions. AB - Data collected from four distinct canopy positions from each of 39 Abies balsamea (L.) Miller trees were used to construct models to describe the cumulative leaf area distribution within the crown and to predict the needle mass of individual branches, the average branch angle, branch diameter, branch length, and crown radius per whorl, and the average number of living branches per whorl. We tested the hypotheses that regression models are equal among canopy positions and that a model to predict branch needle mass is valid at the northern and southern extremes of the central climatic zone of Maine. Canopy position had an effect on the models constructed to predict needle mass, branch angle, branch diameter, branch length, crown radius, and the number of living branches per whorl. However, compared with an expanded model that incorporated parameters calculated for each crown class, there was only a small loss in model precision when a general model constructed from data pooled from all crown classes was used to predict needle mass, branch angle, and branch diameter. Regression equations unique to each crown class were needed to predict crown shape and leaf area distribution in the crown satisfactorily. Our branch needle mass model, which was constructed from data collected at the southern extreme of the central climatic zone of Maine, consistently underestimated needle branch mass when applied to the northern extreme of the central climatic zone. PMID- 14759877 TI - Effect of heat on cambial reactivation during winter dormancy in evergreen and deciduous conifers. AB - Responses of cambium to warming were recorded three times (December 14-27, 1990, January 18-February 3 and February 27-March 13, 1991) on 14-year-old Cryptomeria japonica D. Don and four times (December 12-26, 1990, January 18-February 2, February 26-March 12 and March 28-April 13, 1991) on 27-year-old Larix leptolepis Gord., during a period of winter cambial dormancy. Stem surfaces at breast height, mid-tree height and the crown base were warmed to 25-30 degrees C for 2 weeks. After heat treatment, cambia in the treated regions and in untreated regions 1 m above each treated area were examined by optical and transmission electron microscopy (TEM). In C. japonica, heat treatment often resulted in cambial reactivation in the treated regions, and this response to heat gradually increased as the dormant season passed from winter to spring. Conversely, in L. leptolepis, no cell division was observed in the cambial region of warmed stems until natural resumption of cambial activity, which occurred after bud break. PMID- 14759878 TI - Leaf physiology, production, water use, and nitrogen dynamics of the grassland invader Acacia smallii at elevated CO(2) concentrations. AB - Invasion by woody legumes can alter hydrology, nutrient accumulation and cycling, and carbon sequestration on grasslands. The rate and magnitude of these changes are likely to be sensitive to the effects of atmospheric CO(2) enrichment on growth and water and nitrogen dynamics of leguminous shrubs. To assess potential effects of increased atmospheric CO(2) concentrations on plant growth and acquisition and utilization of water and nitrogen, seedlings of Acacia smallii Isely (huisache) were grown for 13 months at CO(2) concentrations of 385 (ambient), 690, and 980 micro mol mol(-1). Seedlings grown at elevated CO(2) concentrations exhibited parallel declines in leaf N concentration and photosynthetic capacity; however, at the highest CO(2) concentration, biomass production increased more than 2.5-fold as a result of increased leaf photosynthetic rates, leaf area, and N(2) fixation. Measurements of leaf gas exchange and aboveground biomass production and soil water balance indicated that water use efficiency increased in proportion to the increase in atmospheric CO(2) concentration. The effects on transpiration of an accompanying decline in leaf conductance were offset by an increase in leaf area, and total water loss was similar across CO(2) treatments. Plants grown at elevated CO(2) fixed three to four times as much N as plants grown at ambient CO(2) concentration. The increase in N(2) fixation resulted from an increase in fixation per unit of nodule mass in the 690 micro mol mol(-1) CO(2) treatment and from a large increase in the number and mass of nodules in plants in the 980 micro mol mol(-1) CO(2) treatment. Increased symbiotic N(2) fixation by woody invaders in response to CO(2) enrichment may result in increased N deposition in litterfall, and thus increased productivity on many grasslands. PMID- 14759879 TI - Gas exchange and water relations of Fraxinus americana affected by flurprimidol. AB - Effects of flurprimidol on plant water relations and leaf gas exchange were investigated in one-year-old white ash (Fraxinus americana L.) seedlings subjected to soil water deficits. Flurprimidol (20 mg kg(-1) of soil equivalent) was applied to the soil surface of pot-grown seedlings after shoot growth was completed. Two months after flurprimidol application, water was withheld from one half of the seedlings. Leaf water relations and gas exchange parameters were measured 5, 7, 10, 14, 18 and 22 days after withholding water. Under both irrigated and nonirrigated conditions, flurprimidol treatment resulted in reduced net CO(2) assimilation rate and transpirational water loss of seedlings as a result of decreased stomatal conductance. Consequently, flurprimidol-treated seedlings had higher leaf water potential and relative water content than untreated seedlings. Nonirrigated flurprimidol-treated seedlings also had greater turgor and sap osmolality and lower osmotic potential at full turgor than seedlings in the other treatments, indicating that flurprimidol increased osmotic adjustment. Under water-stress conditions, water use efficiency was lower and gas exchange efficiency was higher in flurprimidol-treated seedlings than in untreated seedlings, suggesting that flurprimidol treatment enhances survival of plants subjected to soil water deficits. PMID- 14759880 TI - Biomechanical and hydraulic determinants of tree structure in Scots pine: anatomical characteristics. AB - The development of anatomical, hydraulic and biomechanical properties in Scots pine (Pinus sylvestris L.) stems aged 7 to 59 years was followed. The hydraulic diameter and length of tracheids increased with age to a maximum at 15 and 35 years, respectively. Number of tracheids per unit of sapwood area decreased with age to a minimum of 500-600 tracheids mm(-2). Variations in specific hydraulic conductivity and Young's modulus of stems were associated with variation in anatomical properties. Over the time sequence considered, hydraulic and mechanical properties were positively related to each other and followed a similar developmental pattern, with no suggestion of a trade-off between the two. For most of the tree's life-cycle, heartwood made only a small contribution to whole-section mechanical stiffness because of its location close to the flexural neutral axis, and because of the presence of juvenile wood. PMID- 14759881 TI - Relationships between gas exchange adaptation of Sitka x interior spruce genotypes and ribosomal DNA markers. AB - Adaptive physiological changes were investigated in seven populations of Sitka (Picea sitchensis (Bong.) Carr.) x interior spruce (P. glauca (Moench) Voss x P. engelmannii Parry ex Engelm.) spanning the Nass-Skeena transition zone in British Columbia, Canada. Each population was represented by an Si rDNA index that was calculated from the relative optical densities on a gel autoradiogram of five ribosomal DNA bands characteristic of Sitka spruce and interior spruce. This index estimates the proportion of the genome contributed by interior spruce. Physiological adaptations were assessed by gas exchange parameters measured under both well-watered and drought conditions. Under well-watered conditions, Sitka spruce populations had higher maximal photosynthesis at saturating light and ambient CO(2), higher quantum yield at the light compensation point, and higher dark respiration than interior spruce populations. Sitka spruce populations also reached maximal photosynthesis at lower photosynthetically active radiation and higher CO(2) concentrations, and had higher stomatal densities that resulted in lower stomatal limitations to photosynthesis than interior spruce populations. In contrast, interior spruce populations exhibited greater drought tolerance than Sitka spruce populations. Their gas exchange rates declined at a slower rate in response to drought. They maintained higher gas exchange rates in response to moderate to severe drought (predawn plant water potentials = -1.5 MPa), and their photosynthetic rates recovered faster when they were rewatered after exposure to drought. Comparison of the seven populations indicated that physiological parameters were significantly related to the Si rDNA index. An increase in Si rDNA index was associated with proportional changes in physiological measurements, suggesting that genetic interchange among species with contrasting ecological adaptations can enhance the environmental adaptation of natural populations. PMID- 14759882 TI - Effects of light quality on growth and N accumulation in birch seedlings. AB - We studied the effects of light quality and nutrient supply on growth and nitrogen accumulation in silver birch (Betula pendula Roth) seedlings to test three hypotheses: (1) growth of birch seedlings is sensitive to changes in light quality; (2) the response of birch seedling growth to light quality depends on nutrient supply; and (3) assimilation and allocation of nitrogen by birch seedlings are affected by light quality. The two light regimes simulated the spectral quality of sunlight and shadelight, but did not differ in photosynthetic photon flux density, and the two nutrient supply regimes differed in the rate of supply, but not in the composition, of mineral nutrients. Accumulation and allocation of dry weight and nitrogen were strongly affected by nutrient supply regime, but light quality had little effect. During the first 15 days of the experiment, the largest effect of light quality was on height growth, which was greater in seedlings in simulated shadelight than in seedlings in simulated sunlight. Light quality had little effect on dry weight and nitrogen allocation to the stem during this period. However, at the end of the experiment (Day 29), there was an increase in N concentration per unit dry weight in leaves and stems of seedlings in the simulated shadelight plus high nutrient supply treatment. PMID- 14759883 TI - Influence of photosynthetic photon flux density on growth and transpiration in seedlings of Fagus sylvatica. AB - Beech seedlings (Fagus sylvatica L.) were grown in various combinations of three photosynthetic photon flux densities (PPFD, 0.7, 7.3 or 14.5 mol m(-2) day(-1)) for two years in a controlled environmental chamber. Dry mass of leaves, stem and roots, leaf area and number of leaves, and unit leaf rate were affected by both previous-year and current-year PPFD. Number of shoots and length of the main shoot were affected by previous-year PPFD but not by current-year PPFD. Number of leaves per shoot did not change with PPFD, whereas leaf dry mass/leaf area ratio was mainly affected by current-year PPFD. During the first 10 days that newly emerged seedlings were grown at a PPFD of 0.7 or 14.5 mol m(-2) day(-1), transpiration rate per unit leaf area declined. Thereafter, transpiration increased to a constant new rate. Transpiration rate per seedling was closely related to leaf area but the relationship changed with time. In two-year-old seedlings grown at various PPFD combinations of 0.7, 7.3 and 14.5 mol m(-2) day( 1) during Years 1 and 2, leaf area and transpiration rate per seedling were closely correlated at Weeks 7 and 11 after bud burst. Weak correlations were found between root dry mass and transpiration rate per seedling. During Year 2, transpiration rate per leaf area was higher at a particular PPFD in seedlings grown at a previous-year PPFD of 0.7 mol m(-2) day(-1) than in seedlings grown at a previous-year PPFD of 14.5 mol m(-2) day(-1). After transfer of two-year-old seedlings at the end of the experiment to a new PPFD (7.3 or 14.5 mol m(-2) day( 1)) for one day, transpiration rates per leaf area, measured at the new PPFD, were correlated with leaf area and root dry mass, irrespective of former PPFD treatment. PMID- 14759884 TI - Estimating stand water use of large mountain ash trees and validation of the sap flow measurement technique. AB - Mountain ash (Eucalyptus regnans F.J. Muell.) forest catchments exhibit a strong relationship between stand age and runoff, attributed inter alia to differences in tree water use. However, the tree water use component of the mountain ash forest water balance is poorly quantified. We have used the sap flow technique to obtain estimates of daily water use in large mountain ash trees. First, the sap flow technique was validated by means of an in situ cut tree experiment. Close agreement was obtained between the sap flow estimate of water use and the actual uptake of water by the tree from a reservoir. Second, we compared the variability in sap velocity between a symmetric and an asymmetric tree by using multiple sap flow loggers. In the symmetric tree, velocity was fairly uniform throughout the xylem during the day, indicating that accurate sap flow estimates can be obtained with a minimal number of sampling points. However, large variations in sap velocity were observed in the asymmetric tree, indicating that much larger sampling sizes are required in asymmetric stems for an accurate determination of mean sap velocity. Finally, we compared two procedures for scaling individual tree sap flow estimates to the stand level based on stem diameter and leaf area index measurements. The first procedure was based on a regression between stem diameter and tree water use, developed on a small sample of trees and applied to a stand-level census of stem diameter values. Inputs to the second procedure were tree water use and leaf area of a single tree and the leaf area index of the stand. The two procedures yielded similar results; however, the first procedure was more robust but it required more sampling effort than the second procedure. PMID- 14759885 TI - Gas exchange of the lowest branches of young Scots pine: a cost-benefit analysis of seasonal branch carbon budget. AB - A cost-benefit approach was developed to analyze the carbon budget of the lowest Scots pine (Pinus sylvestris L.) branches subject to abscission. In addition to within-branch growth and respiratory costs, the budget included an estimation of a branch's share of the maintenance respiration of the stem and root. A branch was considered productive if the budget was positive. Foliar gas exchange and woody-tissue respiration were non-destructively measured at monthly intervals during the growing season on the six lowest branches of 10-year-old Scots pine trees, to the moment when the branches died naturally. Photosynthetic light response and temperature response of respiration, together with measurements of canopy light conditions and meteorological data, were used to calculate seasonal carbon budgets for the branches. Maintenance respiration of stems and roots was estimated from published data. All but one of the branches studied were found to be nonproductive over the growing season. Following a decrease in photosynthetic capacity in July, the cumulative budget became negative and the branches died, indicating that a negative carbon budget corresponds with the onset of abscission of the lowest branches. PMID- 14759886 TI - Photosynthetic decline and pigment loss during autumn foliar senescence in western larch (Larix occidentalis). AB - We measured needle pigment content and photosynthetic rates of 1-year-old western larch (Larix occidentalis Nutt.) during autumn foliar senescence. Chlorophyll (Chl) and carotenoid (xanthophyll + b-carotene) contents of needles declined 11 and 17%, respectively, before CO(2) assimilation rate began to decline. Chlorophyll a/b ratio, Chl/carotenoid ratio, photochemical efficiency (F(v)/F(m)), and photochemical quenching did not begin to decline until late in senescence. Internal CO(2)/ambient CO(2) did not change during needle yellowing. In seedlings in warmed soil (average 3 degrees C above natural conditions), the decline in needle chlorophyll content was delayed by 10 days and the decline in CO(2) assimilation rate was delayed by 5 days, compared with seedlings in soil at ambient temperature. In seedlings exposed to an extended 16-h photoperiod, the decline in needle chlorophyll content was delayed by 32 days, and the decline in CO(2) assimilation rate was delayed by 21 days, compared with seedlings exposed to natural day lengths. In addition to delaying the onset of needle senescence, the treatments affected the sequence of events during senescence. Differences among treatment groups provide evidence that the onset of pigment loss and photosynthetic decline and the sequence of events during needle senescence are affected by soil temperature and day length. PMID- 14759887 TI - Nutrient contents and concentrations in relation to growth of Picea abies and Fagus sylvatica along a European transect. AB - Mineral nutrition of Norway spruce (Picea abies (L.) Karst.) and beech (Fagus sylvatica L.) was investigated along a transect extending from northern Sweden to central Italy. Nitrogen (N) concentrations of needles and leaves in stands growing on acid soils did not differ significantly between central Italy and southern Sweden (1.0 +/- 0.1 mmol N g(-1) for needles and 1.9 +/- 0.14 mmol N g( 1) for leaves). In both species, foliar N concentrations were highest in Germany (1.2 mmol N g(-1) for needles and 2.0 mmol N g(-1) for leaves) and decreased by 50% toward northern Sweden (0.5 mmol N g(-1)). Both species showed constant S/N and P/N ratios along the transect. Calcium, K and Mg concentrations generally reflected local soil conditions; however, Mg concentrations reached deficiency values in Germany. Leaf area per unit dry weight varied significantly along the transect with lowest values for Norway spruce recorded in northern Sweden and Italy (3.4 m(2) kg(-1)) and a maximum in central Europe (4.7 m(2) kg(-1)). A similar pattern was observed for beech. Despite the low variation in foliar N concentrations on the large geographic scale, local and regional variations in N concentrations equalled or exceeded the variation along the entire continental transect. Furthermore, nutrient contents (i.e., nutrient concentration x dry weight per needle or leaf) showed a greater variation than nutrient concentrations along the transect. Nitrogen contents of Norway spruce needles reached minimum values in northern Sweden (2.4 micro mol N needle(-1)) and maximum values in Denmark (5.0 micro mol N needle(-1)). The N content of beech leaves was highest in Denmark (242 micro mol N leaf(-1)). At the German site, foliar N content rather than N concentration reflected the seasonal dynamics of foliar growth and N storage of the two species. During foliage expansion, there was an initial rapid increase in N content and a decrease in N concentration. This pattern lasted for about 2 weeks after bud break and was followed by 6 weeks during which dry weight and N content of the foliage increased, resulting in a further decrease in N concentration. During summer, dry weight and N content of mature needles of Norway spruce increased further to reach a maximum in autumn, whereas N concentration remained constant. In spring, reallocation of N from 1- and 2-year-old needles was 1.5 and 1.0 micro mol N needle(-1), respectively. This remobilized N was a major source of N for the development of new needles, which had an N content of 1.5 micro mol N needle(-1) after bud break. The seasonal remobilization of N from old foliage decreased with increasing needle age. Needle N content and dry weight decreased progressively with age (1 micro mol N needle( 1) between age classes 2 and 5), whereas N concentrations remained constant. For Norway spruce, annual stemwood production was correlated with needle N content but not with foliar N concentration or with the total amount of N in the canopy. Interspecific and geographical differences in plant nutrition are discussed on the basis of competitive demands for C and N between growth of foliage and wood. PMID- 14759888 TI - In vitro regeneration of plantlets from mature embryos of Pinus ayacahuite. AB - A plantlet regeneration protocol was developed for Pinus ayacahuite var. ayacahuite (Ehrenb.). Embryos from mature seeds from ten provenances were cultured in a 16-h photoperiod for 3 days on a medium containing 30 mM sucrose and 0.7% agar. Cotyledons from these embryos were subcultured onto MCM medium (Bornman 1983) supplemented with 50 micro M N(6)-benzyladenine and 90 mM sucrose for 2 weeks. Bud development and shoot elongation were maximized by subculturing the explants on half strength AE medium (von Arnold and Ericksson 1981), supplemented with 60 mM sucrose and 0.05% activated charcoal every 30 days. Seed source had a significant effect on the responses of the embryos to the bud induction protocol. For the provenance with the best response to bud induction, about 79% of the cultured cotyledons formed buds, and each cotyledon formed a mean of 9.1 buds, so that about 70 shoots could be induced from each seed. The best rooting response (40% rooting) was obtained by treating the shoots for 8 h with 100 micro M naphthalene acetic acid. PMID- 14759889 TI - Genotypic variation in drought tolerance of poplar in relation to abscisic acid. AB - We investigated effects of water stress and external abscisic acid (ABA) supply on shoot growth, stomatal conductance and water status in 1-year-old cuttings of a drought-sensitive poplar genotype Populus x euramericana cv. I-214 (Italica) and a drought-tolerant genotype P. 'popularis 35-44' (popularis). Populus popularis was more productive and maintained higher leaf water potentials throughout the drought treatment than cv. Italica. Supply of ABA to the xylem sap caused a greater decline in growth and more leaf abscission in shoots of cv. Italica than in shoots of P. popularis. Immediately after initiation of the drought treatment in P. popularis, the ABA concentration ([ABA]) of the xylem increased rapidly and stomatal conductance declined; however, stomatal conductance had returned to control values by the third day of the drought treatment, coincident with a gradual decline in xylem [ABA]. In contrast, xylem [ABA] of cv. Italica initially increased more slowly than that of P. popularis in response to the drought treatment, but the increase continued for 3 days at which time a tenfold increase in xylem [ABA] was observed that was followed by abscission of more than 40% of the leaves. We conclude that sensitivity of poplar roots to variation in soil water content varies by clone and that a rapid short term accumulation of ABA in shoots in response to water stress may contribute to drought tolerance. PMID- 14759890 TI - Carry-over effects of ozone on root growth and carbohydrate concentrations of ponderosa pine seedlings. AB - Ozone exposure decreases belowground carbon allocation and root growth of plants; however, the extent to which these effects persist and the cumulative impact of ozone stress on plant growth are poorly understood. To evaluate the potential for plant compensation, we followed the progression of ozone effects, with particular emphasis on the development of new roots. Ponderosa pine (Pinus ponderosa Dougl. ex Laws.) seedlings were exposed to ozone for 2 years. Following removal of the seedlings from ozone, root growth was assessed to characterize the carry-over effects on new root production, and carbohydrate concentrations were measured to determine if allocation strategies differed among ozone treatments. Four months after removal from ozone, dormant seedlings had significantly lower starch concentrations in stems, coarse roots and fine roots than control seedlings. Following root flushing, starch concentrations in all seedlings decreased, with ozone-treated seedlings containing significantly less starch, sucrose, fructose, glucose and total monosaccharides than control seedlings. There was some evidence that stem starch was mobilized to compensate partially for the lower concentrations of root starch in ozone-treated seedlings; however, there was significantly less new root production in seedlings previously exposed to ozone for 2 years than in control seedlings. Early senescence of older needle age classes, perhaps resulting in inadequate available photosynthate, may be responsible for the reduction in new root production during the year following exposure to ozone. Stored carbohydrate reserves, which were depleted in seedlings previously exposed to ozone, were insufficient to compensate for the ozone induced reduction in canopy photosynthate. We conclude that there are carry-over effects of ozone exposure on ponderosa pine seedlings, including an enhanced potential for seedling susceptibility to other stresses even in respite years when ozone concentrations are low. PMID- 14759891 TI - Leaf nutrient variation in mature carob (Ceratonia siliqua) trees in response to irrigation and fertilization. AB - Seasonal variations in leaf nitrogen, phosphorus and potassium concentrations were studied in a mature carob (Ceratonia siliqua L. cv "Mulata") orchard subjected to a 4-year irrigation and fertilization experiment. Three irrigation regimes (0, 50 and 100%), based on the evaporation values obtained from a class A pan, were tested in combination with two nitrogen (N) supply regimes in which 21 kg ha(-1) year(-1) (low-N) and 63 kg ha(-1) year(-1) (high-N) were supplied as ammonium nitrate. Leaf nitrogen concentration increased throughout the experiment, independently of treatments. There were no significant differences in leaf N concentration between trees in the high-N and low-N treatments. Irrigation regimes had no effect on leaf mineral concentration but influenced the amount of leaves shed and slightly modified the pattern of leaf shedding that occurred during the summer drought period. Nutritional balances between N and P and N and K were both closely and significantly correlated. Potassium was translocated from leaves to fruits during spring, independently of treatments. Severe water stress periods occurring during spring or autumn induced shedding of leaves leading to nutrient mobilization. Nutrient retranslocation during these drought periods may represent an adaptive mechanism. Nitrogen retranslocation was higher for trees in the high-N treatments than for trees in the low-N treatments, whereas phosphorus retranslocation was independent of the irrigation and fertilization treatments. PMID- 14759892 TI - Effects of foliar potassium concentration on morphology, ultrastructure and polyamine concentrations of Scots pine needles. AB - We examined the effects of three foliar potassium concentrations (high, intermediate and low) on the morphology, ultrastructure and polyamine concentrations of current-year and 1- and 2-year-old needles of 30-year-old Scots pine (Pinus sylvestris L.) trees. Foliar K concentration had only a slight effect on needle morphology. The sclerenchyma cell walls were thinner, the xylem area was larger, and the resin ducts were smaller in needles with a low K concentration than in needles with a high or intermediate K concentration. In addition, the bundle sheath cells were collapsed in needles having a low K concentration. The secondary growth of phloem tissue and the mesophyll area were greater in needles with a high or intermediate K concentration than in needles with a low K concentration, possibly indicating greater production of photoassimilates in these trees. At the ultrastructural level, mesophyll cells with enlarged central vacuoles and small vacuoles containing electron-dense material were common in needles having a low K concentration. Enlargement of the central vacuole coincided with an exponential increase in putrescine concentration in needles with a low K concentration, suggesting that the central vacuole may function as a storage site for putrescine. PMID- 14759893 TI - Physiological implications of seasonal variation in membrane-associated calcium in red spruce mesophyll cells. AB - We examined the pattern of seasonal variation in total foliar calcium (Ca) pools and plasma membrane-associated Ca (mCa) in mesophyll cells of current-year and 1 year-old needles of red spruce (Picea rubens Sarg.) and the relationship between mCa and total foliar Ca on an individual plant and seasonal basis. Foliar samples were collected from seedlings and analyzed on 16 dated at 2- to 3-week intervals between June 1994 and March 1995. Concentrations of mCa in current-year needles were more seasonally dynamic and responsive to temporal environmental changes than either mCa concentrations of 1-year-old needles, which were largely stable, or total foliar Ca concentrations in both tissues. In current-year needles, mCa was barely evident in early summer, increased steadily through summer, and then increased dramatically in early fall and surpassed the concentration in 1-year old needles. Coincident with the first severe frost, mCa concentrations in current-year needles declined significantly and subsequently maintained concentrations comparable to those of 1-year-old needles. Following an extended January thaw, which included 5 days of minimum temperatures > 5 degrees C, mCa concentrations of current-year needles temporarily, but significantly, declined. However, there was no change in mCa concentrations of 1-year-old needles or total Ca concentrations of either tissue. Total Ca concentrations were stable through midsummer in both tissues, doubled in late summer, and then were stable in both tissues throughout fall and winter. Total Ca concentrations were consistently higher in 1-year-old than in current-year needles. Correlations between concentrations of mCa and total foliar Ca were consistently low and mostly nonsignificant. Thus, the dominant, but insoluble, extracellular Ca pool reflected in commonly measured total foliar Ca concentrations is not a meaningful surrogate for the physiologically important and labile pool associated with the plasma membrane-cell wall compartment of red spruce mesophyll cells. It is likely that shifts in the critical mCa compartment would not be detected by analysis of total foliar Ca pools. Seasonal changes in mCa concentration seemed to parallel seasonal changes in membrane structure, and possibly the important role of extracellular Ca in transducing messages associated with environmental signals. PMID- 14759894 TI - Distribution of elements along the length of Scots pine needles in a heavily polluted and a control environment. AB - Pollution often causes visible symptoms of foliar injury. The injury is sometimes associated with an increase in the accessibility of toxic elements to plants as a result of acidification of the soil. We investigated the distribution of elements (N, P, K, Ca, Mg, Mn, S, Fe, B, Cu, Zn, Al, F, Pb, Cd, Cr, Ni and Co) in healthy current-year needles of Scots pine (Pinus sylvestris L.) growing at an unpolluted control site and at a site polluted mainly by SO(2), HF and Al(3+) from a fertilizer factory established in 1917. Needles from both sites were sampled before the appearance of visible injury and cut into five sections of equal length (tip, base and three middle parts). The mean concentrations of major nutrients were 20-30% lower in needles at the polluted site than in needles at the control site, whereas the concentrations of aluminum and fluorine were higher in needles at the polluted site. An increase in concentration from needle base to tip was detected for N, Fe, B, and Al at both sites and for Mn only at the polluted site. Fluoride accumulated in the tips of needles only at the polluted site, which could explain the necroses of needle tips at this site. The distribution of elements along the length of the needles was influenced by pollution, element mobility and the distal accumulation of toxic elements. PMID- 14759895 TI - Environmental controls over isoprene emission in deciduous oak canopies. AB - In summer 1992, isoprene emission was measured on intact leaves and branches of Quercus alba (L.) at two heights in a forest canopy. Isoprene emission capacity (measured at 30 degrees C and a photosynthetic photon flux density of 1000 micro mol m(-2) s(-1)) was significantly higher in sun leaves than in shade leaves when expressed on a leaf area basis (51 versus 31 nmol m(-2) s(-1); P < 0.01). Because leaf mass per unit area (LMA, g m(-2)) was higher in sun leaves than in shade leaves, emissions of sun and shade leaves expressed on a dry mass basis did not differ significantly (99 versus 89 micro g C g(DW) (-1) h(-1); P = 0.05). Similar measurements in 1995 were consistent with the 1992 data, but data from leaves in more shaded locations demonstrated that isoprene emission capacity decreased with decreasing growth irradiance, irrespective of units of expression. Isoprene emission capacity in leaves of Q. coccinea Muenchh. and Q. velutina Lam. also declined steeply with canopy depth. Emission capacity, on a dry mass basis, showed no obvious pattern with canopy position in Q. prinus L. There was no difference in the temperature response of sun versus shade leaves of Q. alba, but shade leaves exhibited a greater quantum efficiency and saturated at lower irradiance than sun leaves. Rates of isoprene emission measured on branches of Q. alba were approximately 60% of those measured on individual leaves, as a result of self-shading within branch enclosures. It is recommended that within-canopy variation in isoprene emission capacity be incorporated into regional emission models. PMID- 14759896 TI - Control of longitudinal and cambial growth by gibberellins and indole-3-acetic acid in current-year shoots of Pinus sylvestris. AB - We investigated the involvement of gibberellins (GAs) and indole-3-acetic acid (IAA) in the control of longitudinal and cambial growth in current-year shoots of Pinus sylvestris L. Elongating terminal shoots, located at the apex of previous year (1-year-old) branches in the uppermost whorl on the main stem, were variously decapitated (apical 5 to 10 mm removed), defoliated (all developing needle fascicles removed) and treated with endogenous GA(4/7) or IAA, or both. Shoot length and the radial widths of xylem and phloem were measured, and the concentrations of GA(1), GA(3), GA(4), GA(9) and IAA in the stem were determined by combined gas chromatography-mass spectrometry with deuterated GAs and [(13)C(6)]-IAA as internal standards. Decapitation decreased the production of xylem and phloem and the IAA concentration, but did not alter either longitudinal growth or the concentrations of GAs. Defoliation markedly inhibited shoot elongation, as well as cambial growth, and reduced the concentrations of GA(1), GA(3), GA(4), GA(9) and IAA. Application of GA(4/7) to defoliated shoots promoted longitudinal growth and phloem production, without affecting xylem production or IAA concentration. Application of GA(4/7) and IAA together to decapitated + defoliated shoots increased shoot elongation, xylem and phloem production and IAA concentration, whereas applying either substance alone had a smaller effect or none at all. We conclude that, for elongating current-year shoots of Pinus sylvestris, (1) both the shoot apex and the developing needle fascicles are major sources of the IAA present in the stem, whereas stem GAs originate primarily in the needle fascicles, (2) GAs and IAA are required for both shoot elongation and cambial growth, and (3) GAs act directly in the control of shoot growth, rather than indirectly through affecting the IAA concentration. PMID- 14759897 TI - Acclimation to low irradiance in Picea abies: influences of past and present light climate on foliage structure and function. AB - Leaf retention time increases with decreasing irradiance, providing an effective way of amortizing the costs of foliage construction over time. To elucidate the physiological mechanisms underlying this dependence, I studied needle life span, morphology, and concentrations of carbon, nitrogen and nonstructural carbohydrates along a gradient of relative irradiance in understory trees of Picea abies (L.) Karst. Maximum needle life span was greater in shaded trees than in sun-exposed trees. However, irrespective of irradiance, needles with maximum longevity were situated in the middle rather than the bottom of the canopy, suggesting that needle life span is determined by the irradiance to which needles are exposed during their primary growth. Morphology and chemistry of current-year needles were adapted to prevailing light conditions. Current-year needles exposed to high irradiances had greater packing of foliar biomass per unit area than shaded needles, whereas shaded needles maximized foliar area to capture more light. Nitrogen concentrations were higher in shaded needles than in sun-exposed needles. This nitrogen distribution pattern was related to the high nitrogen cost of light interception and was assumed to improve light absorptance per needle mass of shaded needles. In contrast, in both 1- and 2-year-old needles, morphology was independent of prevailing light conditions; however, needle nitrogen concentrations were adjusted toward more effective light interception in 2-year-old foliage but not in 1-year-old foliage, indicating that acclimation of sun-adapted needles to shading takes more than one year. At the same time, needle aging was accompanied by accumulation of nonstructural carbohydrates (NSC), and increasing concentrations of needle carbon, suggesting a shift in the balance between photosynthesis and photosynthate export. The accumulation of NSC and carbon resulted in a dilution of the concentrations of other needle chemicals and explained the decline in needle nitrogen concentrations with increasing age. Thus, although morphological inadequacy to low light availabilities may partly be compensated for by modifications in needle chemistry, age-related changes in needle stoichiometric composition progressively lessen the potential for acclimation to low irradiance. A conceptual model, advanced to explain how environmental factors and age-related changes in the activities of needle xylem and phloem transport affect needle longevity, predicted that adaptation of needle morphology to irradiance during the primary growth period largely determines the fate of needles during subsequent tree growth and development. PMID- 14759898 TI - Sources of N for leaf growth in a high-density apple (Malus domestica) orchard irrigated with ammonium nitrate solution. AB - Elstar apple trees (Malus domestica Borkh.) on M.9 rootstock received either 5 or 35 g N tree(-1) year(-1) during the first two growing seasons after planting, applied as Ca(NO(3))(2) on a daily basis for nine weeks through a drip irrigation system. During the third growing season (1994), all trees were treated with 20 g N tree(-1) year(-1) as (15)NH(4) (15)NO(3) with applications starting on April 22 and continuing for 10 weeks. Soil solution nitrate-N and ammonium-N were monitored weekly with suction lysimeters located 30 cm beneath the drip emitters. Spur and shoot leaves were sampled intensively from full bloom to the end of rapid shoot growth. During the period of nitrogen application, soil solution nitrate-N and ammonium-N were relatively constant, at about 24 and 1.0 mg l(-1) respectively. Growth of the spur leaves was completed by one week after full bloom (May 12), whereas biomass of the shoot leaves increased until mid-June. Nitrogen for growth of the spur leaves was supplied mainly from remobilization, which was dependent on previous N supply. Accumulation of fertilizer N in spur leaves was independent of previous N treatments and continued until the end of the monitoring period (June 24), but contributed only 13% to total spur leaf N. Nitrogen for shoot leaf growth was independent of previous N treatments and was initially supplied primarily by remobilization, but by the end of extension growth, fertilizer N contributed 48% to total shoot leaf N. Linear increases in leaf N uptake throughout the period of rapid shoot growth and the large contribution of fertilizer N to total shoot leaf N were attributed to the constant supply of N available in the root zone through daily N fertilization. PMID- 14759899 TI - Effect of crop load on fruiting and leaf photosynthesis of 'Braeburn'/M.26 apple trees. AB - Four-year-old apple (Malus x domestica Borkh.) trees cv. 'Braeburn' on M.26 rootstock were thinned at full bloom to establish six crop loads ranging from a heavy crop to a deflowered treatment. At harvest, mean yield per tree varied from 0 to 38 kg and mean fruit weight ranged from 225 g in the heaviest cropping treatment to 385 g in the lightest cropping treatment. Light cropping resulted in a significant advance in fruit maturity as indicated by background color, starch/iodine score and soluble solids. There were small differences in leaf photosynthetic rate among the treatments when shoot growth was active. However, in early January, coincident with cessation of shoot growth and maximum rate of accumulation of fruit weight, leaf assimilation rate was reduced by as much as 65% on the deflowered trees compared to the trees carrying the heaviest crop. Leaf assimilation rate showed a curvilinear response to crop load at this time, with little increase in leaf assimilation when crop load exceeded 12 fruit m(-2) leaf area. PMID- 14759900 TI - Leaf conductance and CO(2) assimilation of Larix gmelinii growing in an eastern Siberian boreal forest. AB - In July 1993, we measured leaf conductance, carbon dioxide (CO(2)) assimilation, and transpiration in a Larix gmelinii (Rupr.) Rupr. ex Kuzen forest in eastern Siberia. At the CO(2) concentration of ambient air, maximum values (mean of 10 highest measured values) for CO(2) assimilation, transpiration and leaf conductance for water vapor were 10.1 micro mol m(-2) s(-1), 3.9 mmol m(-2) s(-1) and 365 mmol m(-2) s(-1), respectively. The corresponding mean values, which were much lower than the maximum values, were 2.7 micro mol m(-2) s(-1), 1.0 mmol m( 2) s(-1) and 56 mmol m(-2) s(-1). The mean values were similar to those of Vaccinium species in the herb layer. The large differences between maximum and actual performance were the result of structural and physiological variations within the tree crowns and between trees that reduced maximum assimilation and leaf conductance by about 40 and 60%, respectively. Thus, maximum assimilation and conductance values averaged over the canopy were 6.1 micro mol m(-2) s(-1) and 146 mmol m(-2) s(-1), respectively. Dry air caused stomatal closure, which reduced assimilation by an additional 26%. Low irradiances in the morning and evening had a minor effect (-6%). Daily canopy transpiration was estimated to be 1.45 mm day(-1), which is higher than the value of 0.94 mm day(-1) measured by eddy covariance, but similar to the value of 1.45 mm day(-1) calculated from the energy balance and soil evaporation, and less than the value of 2.1 mm day(-1) measured by xylem flux. Daytime canopy carbon assimilation, expressed on a ground area basis, was 0.217 mol m(-2) day(-1), which is higher than the value measured by eddy flux (0.162 mol m(-2) day(-1) including soil respiration). We discuss the regulation of leaf gas exchange in Larix under the extreme climatic conditions of eastern Siberia (temperature > 35 degrees C and vapor pressure deficit > 5.0 kPa). PMID- 14759901 TI - Growth, leaf anatomy, and physiology of Populus clones in response to solar ultraviolet-B radiation. AB - We compared the physiological and morphological responses of rooted cuttings of Populus trichocarpa Torr. & Gray and P. trichocarpa x P. deltoides Bartr. ex Marsh. grown in either near-ambient solar ultraviolet-B (UV-B; 280-320 nm) radiation (cellulose diacetate film) or subambient UV-B radiation (polyester film) for one growing season. Midday biologically effective UV-B radiation was 120.6 and 1.6 mJ m(-2) s(-1) under the cellulose diacetate and polyester films, respectively. Gas exchange, leaf chlorophyll, light harvesting efficiency of photosystem II, and foliar UV-B radiation-absorbing compounds (i.e., flavonoid derivatives) were measured in expanding (leaf plastochron index (LPI) 5), nearly expanded (LPI 10), and fully expanded mature (LPI 15) leaves of intact plants of plastochron index 30 to 35. Plants were then harvested and height, diameter, biomass allocation and leaf anatomical attributes determined. Net photosynthesis, transpiration, and stomatal conductance were significantly greater in mature leaves exposed to subambient UV-B radiation than in mature leaves exposed to near ambient UV-B radiation. Concentrations of UV-B radiation-absorbing compounds (measured as absorbance of methanol-extracts at 300 nm) were significantly greater in mature leaves exposed to near-ambient UV-B radiation than in mature leaves exposed to subambient UV-B radiation. The UV-B radiation treatments had no effects on chlorophyll content or intrinsic light harvesting efficiency of photosystem II. Height, diameter, and biomass were not significantly affected by UV-B radiation regime in either clone. Leaf anatomical development was unaffected by UV-B radiation treatment in P. trichocarpa x P. deltoides. For P. trichocarpa, leaf anatomical development was complete by LPI 10 in the near-ambient UV-B radiation treatment, but continued through to LPI 15 in the subambient UV-B radiation treatment. Mature leaves of P. trichocarpa were thicker in the subambient UV-B radiation treatment than in the near-ambient UV-B radiation treament as a result of greater development of palisade parenchyma tissue. We conclude that exposure to near-ambient UV-B radiation for one growing season caused shifts in carbon allocation from leaf development to other pools, probably including but not limited to, UV-B absorbing compounds. This reallocation curtailed leaf development and reduced photosynthetic capacity of the plants compared with those in the subambient UV-B radiation treatment and may affect growth over longer periods of exposure. PMID- 14759902 TI - Simulating the response of mature yellow poplar and loblolly pine trees to shifts in peak ozone periods during the growing season using the TREGRO model. AB - Multiple TREGRO simulations were conducted with meteorological data files containing different growing season peak ozone (O(3)) episodes at O(3) exposures of 1.0 and 2.0 x ambient O(3) to assess the relationship between O(3) response and the phenology of mature yellow poplar (Liriodendron tulipifera L.) and loblolly pine (Pinus taeda L.) trees. Regardless of O(3) exposure and peak O(3) episode occurrence, a peak O(3) episode in August caused the greatest reduction in carbon (C) gain in yellow poplar, whereas a peak O(3) episode in July caused the greatest reduction in C gain of loblolly pine. In both species, timing of the greatest simulated O(3) effect corresponded with the completion of the annual foliage production phenophase. Simulated C gain of yellow poplar (total tree, coarse root, and total nonstructural carbohydrate) was reduced by O(3) to a greater extent than the corresponding compartments in loblolly pine, but the opposite was true for fine roots. This differential sensitivity to O(3) reflects the fact that both C assimilation and the O(3) response of the species were parameterized according to observed field measurements of each species. The differential sensitivity to O(3) of these species may have long-term implications for species composition in southeastern USA forests. PMID- 14759903 TI - Influence of environmental and plant factors on canopy photosynthesis and transpiration of apple trees. AB - We estimated carbon and water flows, canopy conductance and the assimilation/transpiration ratio of fruiting and non-fruiting apple trees grown in the field, from daily gas exchange measurements taken during the summer with a whole-canopy enclosure device. The relationships between photosynthetic and transpirational responses and environmental conditions were also investigated, as well as the role of canopy conductance in controlling carbon dioxide and water vapor exchange. Light-saturated net photosynthetic rates, which were higher for the fruiting canopy than for the non-fruiting canopy, showed a general decrease in the afternoon, particularly for the non-fruiting canopy, compared with rates in the morning. When light was not limiting, the afternoon decrease in net photosynthesis appeared to be regulated more by non-stomatal factors than by changes in canopy conductance. Canopy conductance, which was higher for the fruiting canopy than for the non-fruiting canopy, may actively regulate photosynthetic activity and may also be modulated by feedback control in response to assimilation capacity. We conclude that adjustments in canopy conductance, which were partially dependent on the vegetative-reproductive status of the tree, control the equilibrium between photosynthesis and transpiration. We also demonstrated that whole-canopy chambers can be used to estimate photosynthetic and transpirational responses thereby overcoming the difficulty of scaling these physiological responses from the leaf to the whole-canopy level. PMID- 14759904 TI - Nutrition, xylem cavitation and drought resistance in hybrid poplar. AB - Effects of mineral nutrition on susceptibility to cavitation were examined in four hybrid poplar clones. Two drought-sensitive and two drought-resistant hybrid clones of black cottonwood (Populus trichocarpa Torr. & Gray) and eastern cottonwood (P. deltoides Bartr.) were grown at three concentrations of nitrogen (N) applied factorially with two concentrations of phosphorus (P) in a greenhouse, and subjected to varying degrees of drought stress before measurement of cavitation and of anatomical features that might affect cavitation. Mean vessel pit pore diameters were 0.132 micro m at low P, and 0.074 micro m at high P, but no other significant effects of mineral nutrition on vessel dimensions were observed. Vessel diameter and specific conductivity were greater in the drought-resistant clones than in the drought-susceptible clones. Drought resistant clones did not reach such low water potentials as drought-sensitive clones during the cavitation induction experiments, suggesting better stomatal and cuticular control of water loss. Scanning electron microscope observations showed less damage to pit membranes, also suggesting greater membrane strength in drought-resistant clones than in drought-sensitive clones. High concentrations of N increased cavitation, whereas high concentrations of P decreased cavitation as measured by both hydraulic flow apparatus and dye perfusion techniques. For one test, cavitation was 48% at high N and low P, but only 28% at high N and high P. We consider that N fertilization may make poplars more susceptible to cavitation on dry sites, but P fertilization may reduce this effect. PMID- 14759905 TI - Effect of elevated carbon dioxide concentration and root restriction on net photosynthesis, water relations and foliar carbohydrate status of loblolly pine seedlings. AB - To determine the effects of CO(2)-enriched air and root restriction on photosynthetic capacity, we measured net photosynthetic rates of 1-year-old loblolly pine seedlings grown in 0.6-, 3.8- or 18.9-liter pots in ambient (360 micro mol mol(-1)) or 2x ambient CO(2) (720 micro mol mol(-1)) concentration for 23 weeks. We also measured needle carbohydrate concentration and water relations to determine whether feedback inhibition or water stress was responsible for any decreases in net photosynthesis. Across all treatments, carbon dioxide enrichment increased net photosynthesis by approximately 60 to 70%. Net photosynthetic rates of seedlings in the smallest pots decreased over time with the reduction occurring first in the ambient CO(2) treatment and then in the 2x ambient CO(2) treatment. Needle starch concentrations of seedlings grown in the smallest pots were two to three times greater in the 2x ambient CO(2) treatment than in the ambient CO(2) treatment, but decreased net photosynthesis was not associated with increased starch or sugar concentrations. The reduction in net photosynthesis of seedlings in small pots was correlated with decreased needle water potentials, indicating that seedlings in the small pots had restricted root systems and were unable to supply sufficient water to the shoots. We conclude that the decrease in net photosynthesis of seedlings in small pots was not the result of CO(2) enrichment or an accumulation of carbohydrates causing feedback inhibition, but was caused by water stress. PMID- 14759906 TI - Analysis of bark proteins in blister rust-resistant and susceptible western white pine (Pinus monticola). AB - We compared bark proteins from four contrasting (blister rust-resistant versus susceptible) half-sib seedling pairs of western white pine (Pinus monticola D. Don). Pooled proteins from resistant and susceptible groups (four trees per group) were separated by two-dimensional gel electrophoresis, silver stained, and analyzed with the aid of a laser scanner interfaced with a computerized gel documentation system. Qualitative and quantitative protein differences were observed between resistant and susceptible groups. The number of proteins unique to a group was greater in the susceptible category than in the resistant category. Biosynthesis of some common proteins was enhanced near lesioned areas of susceptible seedlings. Many proteins shared similar charge and mass characteristics with those of pathogenesis-related (PR) proteins. Two protein bands were isolated and partially characterized by N-terminal amino acid sequencing: a 10.6-kDa band that was selectively enriched in all resistant individuals, and a 26.0-kDa band that was enriched in some susceptible individuals. The significance of these protein differences and the possible use of selected proteins as disease or resistance markers are discussed. PMID- 14759907 TI - Tissue culture responses of explants taken from branch sources with different degrees of juvenility in mature black locust (Robinia pseudoacacia) trees. AB - We studied the in vitro responses of cambial tissue and dormant vegetative buds obtained from top and epicormic branches of three mature black locust (Robinia pseudoacacia L.) trees. Cambial tissues isolated from epicormic branches produced more callus than cambial tissues isolated from top branches, whereas in vitro shoot cultures derived from buds excised from top branches grew faster than those derived from buds excised from epicormic branches. There were no significant differences between the two branch sources in in vitro bud break or shoot multiplication from bud explants or cambial-derived callus tissue, respectively. Furthermore, the top branches, generally considered to be the most mature in a tree, were not recalcitrant in terms of morphogenic capacity compared to epicormic branches. PMID- 14759908 TI - Vertical gradients in photosynthetic gas exchange characteristics and refixation of respired CO(2) within boreal forest canopies. AB - We compared vertical gradients in leaf gas exchange, CO(2) concentrations, and refixation of respired CO(2) in stands of Populus tremuloides Michx., Pinus banksiana Lamb. and Picea mariana (Mill.) B.S.P. at the northern and southern boundaries of the central Canadian boreal forest. Midsummer gas exchange rates in Populus tremuloides were over twice those of the two conifer species, and Pinus banksiana rates were greater than Picea mariana rates. Gas exchange differences among the species were attributed to variation in leaf nitrogen concentration. Despite these differences, ratios of intercellular CO(2) to ambient CO(2) (c(i)/c(a)) were similar among species, indicating a common balance between photosynthesis and stomatal conductance in boreal trees. At night, CO(2) concentrations were high and vertically stratified within the canopy, with maximum concentrations near the soil surface. Daytime CO(2) gradients were reduced and concentrations throughout the canopy were similar to the CO(2) concentration in the well-mixed atmosphere above the canopy space. Photosynthesis had a diurnal pattern opposite to the CO(2) profile, with the highest rates of photosynthesis occurring when CO(2) concentrations and gradients were lowest. After accounting for this diurnal interaction, we determined that photosynthesizing leaves in the understory experienced greater daily CO(2) concentrations than leaves at the top of the canopy. These elevated CO(2) concentrations were the result of plant and soil respiration. We estimated that understory leaves in the Picea mariana and Pinus banksiana stands gained approximately 5 to 6% of their carbon from respired CO(2). PMID- 14759909 TI - Analysis of leaf water relations in leaves of two olive (Olea europaea) cultivars differing in tolerance to salinity. AB - One-year-old rooted cuttings of olive (Olea europaea L. cvs. Frantoio and Leccino) were grown either hydroponically or in soil in a greenhouse. Plants were exposed to NaCl treatments (0, 100, and 200 mM) for 35 days, followed by 30 to 34 days of relief from salt stress to determine whether previously demonstrated genotypic differences in tolerance to salinity were related to water relations parameters. Exposure to high salt concentrations resulted in reductions in predawn water potential (Psi(w)), osmotic potential at full turgor (Psi(piFT)), osmotic potential at turgor loss point (Psi(piTLP)), and relative water content (RWC) in both cultivars, regardless of the growth substrate. Leaf Psi(w) and RWC returned to values similar to those of controls by the end of the relief period. The effect of salinity on Psi(pi) appeared earlier in Leccino than in Frantoio. Values for Psi(piFT) were -2.50, -2.87, and -3.16 MPa for the 0, 100, and 200 mM salt-treated Frantoio plants, respectively, and -2.23, -2.87, and -3.37 MPa for the corresponding Leccino plants. Recovery of Psi(pi) was complete for plants in the 100 mM salt treatment, but not for plants in the 200 mM salt treatment, which maintained an increased pressure potential (Psi(pi)) compared to control plants. Net solute accumulation was higher in Leccino, the salt-sensitive cultivar, than in Frantoio. In controls of both cultivars, cations contributed 39.9 to 42.0% of the total Psi(piFT), mannitol and glucose contributed 27.1 to 30.8%, and other soluble carbohydrates contributed 3.1 to 3.6%. The osmotic contribution of Na(+) increased from 0.1-2.1% for non-treated plants to 8.6-15.5% and 15.6-20.0% for the 100 mM and 200 mM salt-treated plants, respectively. The mannitol contribution to Psi(piFT) reached a maximum of 9.1% at the end of the salinization period. We conclude that differences between the two cultivars in leaf water relations reflect differences in the exclusion capacities for Na(+) and Cl(-) ions. PMID- 14759910 TI - Light availability and photosynthesis of Pseudotsuga menziesii seedlings grown in the open and in the forest understory. AB - The light environment, photosynthetic dynamics and steady-state net photosynthetic rates of lateral branch shoots of Pseudotsuga menziesii var. glauca (Beissn.) Franco seedlings growing in the open and in the forest understory were investigated in situ. Mean incident photosynthetic photon flux density (PPFD) was 702.5 micro mol m(-2) s(-1) on open-grown branches and 52.0 micro mol m(-2) s(-1) on understory-grown branches. Mean daily durations of PPFD greater than 500, 200, and 50 micro mol m(-2) s(-1) were 8.5, 31.5, and 270.3 min, respectively, on understory-grown branches, and 559.1, 700.7, and 803.3 min, respectively, on open-grown branches. Sunflecks accounted for 32.4% of total daily photosynthetically active radiation incident on understory branches. Following 10 min at a PPFD of 50 micro mol m(-2) s(-1), the induction time required for net photosysnthesis to reach 50 and 90% of steady-state rates was shorter at a PPFD of 200 than at a PPFD of 500 micro mol m(-2) s(-1) and shorter in understory-grown branches than in open-grown branches. On a leaf area basis, dark respiration rates of understory-grown branches were lower and net photosynthetic rates were higher than those of open-grown branches exposed to low PPFD. However, at high PPFDs, understory-grown branches had lower photosynthetic rates than open-grown branches. When measurements were expressed on a leaf dry mass basis, there was no difference in dark respiration rates between understory branches and open-grown branches, but net photosynthetic rates of understory branches were equal to or higher than those of open-grown branches at all PPFDs. PMID- 14759911 TI - Changes in ABA and gene expression in cold-acclimated sugar maple. AB - To determine if cold acclimation of sugar maple (Acer saccharum Marsh.) is associated with specific changes in gene expression under natural hardening conditions, we compared bud and root translatable mRNAs of potted maple seedlings after cold acclimation under natural conditions and following spring dehardening. Cold-hardened roots and buds were sampled in January when tissues reached their maximum hardiness. Freezing tolerance, expressed as the lethal temperature for 50% of the tissues (LT(50)), was estimated at -17 degrees C for roots, and at lower than -36 degrees C for buds. Approximately ten transcripts were specifically synthesized in cold-acclimated buds, or were more abundant in cold acclimated buds than in unhardened buds. Cold hardening was also associated with changes in translation. At least five translation products were more abundant in cold-acclimated buds and roots compared with unhardened tissues. Abscisic acid (ABA) concentration increased approximately tenfold in the xylem sap following winter acclimation, and the maximum concentration was reached just before maximal acclimation. We discuss the potential involvement of ABA in the observed modification of gene expression during cold hardening. PMID- 14759912 TI - Responses of Picea, Pinus and Pseudotsuga roots to heterogeneous nutrient distribution in soil. AB - The spatial distribution of plant-available mineral nutrients in forest soils is often highly heterogeneous. To test the hypothesis that local nutrient enrichment of soil leads to increased root proliferation in the nutrient-rich soil zone, we studied the effects of nutrient enrichment on the growth and nutrient concentrations of Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco), Scots pine (Pinus sylvestris L.) and Norway spruce (Picea abies (L.) Karst.) roots. Three year-old seedlings were grown for 9 months in split-root containers filled with nutrient-poor forest mineral soil, with one side supplemented with additional mineral nutrients. Root dry weight and root length in Scots pine and Norway spruce were increased in the nutrient-supplemented soil compared with the nonsupplemented side, whereas root growth in Douglas-fir was unaffected by nutrient enrichment. Of the three species examined, Norway spruce exhibited the highest root and shoot growth and the highest nutrient demand. Specific root length (m g(-1)) and the number of root tips per unit root length were not affected by local nutrient addition in any of the species. Despite increased root growth in Norway spruce and Scots pine in nutrient-supplemented soil, their root systems contained similar nutrient concentrations on both sides of the split-root container. Thus, coniferous trees may respond to local nutrient supply by increased root proliferation, but the response varies depending on the species, and may only occur when trees are nutrient deficient. As a response to local nutrient enrichment, increases in root dry matter or root length may be better indicators of pre-existing nutrient deficiencies in conifers than increases in root nutrient concentrations. PMID- 14759913 TI - Relationship between temperature, respiratory loss of sugar and premature dehardening in dormant Scots pine seedlings. AB - Increased intracellular sugar concentration is an important contributor to the increased cold tolerance of conifers in winter. This study examines the extent to which wintertime respiratory loss of sugars leads to premature dehardening. Two year-old seedlings of Scots pine (Pinus sylvestris L.), grown and cold-hardened in the field, were exposed to different temperature regimes for 16 weeks while dormant. To minimize short-term carry-over effects, after the temperature treatments, all seedlings were conditioned to 5.5 degrees C and watered before the assessment of non-structural carbohydrates and cold tolerance. Needle sugar concentration was decreased by 54, 32, 21 and 9% following treatment at 5.5, 0, 1.5 and -8.5 degrees C, respectively. Sugar concentration did not decrease as much in root tissues as in needles because starch was mobilized in roots. Cold tolerance of needles was analyzed by controlled freezing, and the temperature causing an initial 10% damage (LT(10)) was plotted as a function of needle sugar concentration, revealing a strong, linear relationship. When one-third of the initial sugars had been consumed, LT(10) had increased from -24.5 to -16.5 degrees C, and when one half had been consumed, LT(10) had increased to -10 degrees C. Consequences of these findings for the field performance of conifers are discussed in relation to climatic variation and change. PMID- 14759914 TI - Influences of altered phytohormone use on endogenous ABA and mRNA populations during white spruce (Picea glauca) somatic embryo culture. AB - We examined endogenous abscisic acid [(+)-ABA] amounts and mRNA populations in white spruce (Picea glauca (Moench) Voss) somatic embryos in liquid suspension culture during a pretreatment prior to induction of maturation with exogenous (+) ABA. The pretreatment consisted of removal of 2,4-dichlorophenoxyacetic acid (2,4 D) from the multiplication medium for 7 days. The removal of 2,4-D resulted in a slight increase in endogenous ABA in pretreated tissues compared to nonpretreated tissues at the end of the 7-day pretreatment period (42.4 versus 20.0 pg mg(-1) lyophilized tissue). Altered gene expression patterns were observed as early as one day after the start of the pretreatment, with more than 17 new polypeptides found in pretreated tissues. The influence of pretreatment continued to be observed after tissues were transferred to ABA-containing maturation medium. By comparing mRNA populations in pretreated and nonpretreated tissues cultured on either ABA-containing or ABA-free maturation medium, at least 12 mRNAs were observed to be induced by ABA, among which three polypeptides were ABA inducible only in pretreated tissues. PMID- 14759915 TI - Xylem cavitation and loss of hydraulic conductance in western hemlock following planting. AB - Following planting, western hemlock (Tsuga heterophylla (Raf.) Sarg.) seedlings experience water stress and declining xylem pressure potential (Psi(x)). Low Psi(x) can result in xylem cavitation and embolism formation, causing a decline in hydraulic conductance. This study focused on the relationship between Psi(x), xylem cavitation and transpiration (E) of newly planted seedlings. Leaf specific hydraulic conductance (k(AB)) declined from 0.56 to 0.09 mmol m(-2) s(-1) MPa(-1) over a 9-day period. Stomatal conductance (g(s)) declined from 143.5 to 39.15 mmol m(-2) s(-1) over the same period without an associated change in environmental conditions. A vulnerability profile indicated a 30% loss in hydraulic conductivity when seedlings experienced a Psi(x) between -2.5 and -3.0 MPa. A Psi(x) of -4.0 MPa led to a complete loss of conductivity. We conclude that following planting, western hemlock seedlings often experience Psi(x) values that are low enough to cause xylem cavitation and a decline in k(AB). PMID- 14759916 TI - A unified nomenclature for sap flow measurements. AB - A unified nomenclature for use in heat pulse measurement of sap flow is proposed. This unified nomenclature overcomes fundamental misunderstandings of the physics of heat and sap movement in wood. The nomenclature is also appropriate to other methodologies for sap flow measurement, such as heat balance methods. PMID- 14759917 TI - Traffic safety in the United States. PMID- 14759918 TI - Making vehicles safer. PMID- 14759921 TI - America's choice: reducing tobacco addiction and disease. PMID- 14759922 TI - A silence that kills. PMID- 14759923 TI - Building coalitions for tobacco control and prevention in the 21st century. PMID- 14759924 TI - Appalachian teen smokers: not on tobacco 15 months later. AB - High school smokers from 2 central Appalachian states received the American Lung Association's 10-session Not On Tobacco (N-O-T) program or a 15-minute brief self help intervention. Our study compared the efficacy of N-O-T with that of the brief intervention by examining group differences in the 15-month-postbaseline (12-month-postprogram) smoking quit rates. N-O-T youths had higher overall quit rates. Review of end-of-program (3-month-postbaseline) and 3-month-postprogram (6 month-postbaseline) follow-up data showed state-level differences and positive cessation trends over time, regardless of treatment intensity. Quit rates were lower than rates found in other N-O-T studies of nonrural youths, suggesting that Appalachian youths are a recalcitrant smoking sample. Findings suggest that N-O-T is one option for long-term smoking cessation among rural teens. PMID- 14759925 TI - Hospital smoking bans and their impact. PMID- 14759926 TI - Reversal of misfortune: viewing tobacco as a social justice issue. PMID- 14759927 TI - Out of the ashes: the life, death, and rebirth of the "safer" cigarette in the United States. AB - From 1964 through the early 1980s, both federal and voluntary agencies endorsed the concept of "safer" cigarettes. Beginning in the mid-1980s, several factors, including revelations of tobacco industry malfeasance, the development of nicotine replacement therapy, and the reconceptualization of smoking as a chronic disease, led to "safer" cigarettes being discredited. In the past few years, some public health professionals have begun to reconsider the viability of developing such products. The issue before us is stark: will a commitment to limiting the toll exacted by smoking preclude the tolerance of a product that while not safe may possibly be safer? PMID- 14759928 TI - Preventing 3 million premature deaths and helping 5 million smokers quit: a national action plan for tobacco cessation. AB - In August 2002, the Subcommittee on Cessation of the Interagency Committee on Smoking and Health (ICSH) was charged with developing recommendations to substantially increase rates of tobacco cessation in the United States. The subcommittee's report, A National Action Plan for Tobacco Cessation, outlines 10 recommendations for reducing premature morbidity and mortality by helping millions of Americans stop using tobacco. The plan includes both evidence-based, population-wide strategies designed to promote cessation (e.g., a national quitline network) and a Smokers' Health Fund to finance the programs (through a 2 US dollar per pack excise tax increase). The subcommittee report was presented to the ICSH (February 11, 2003), which unanimously endorsed sending it to Secretary Thompson for his consideration. In this article, we summarize the national action plan. PMID- 14759929 TI - Eliminating tobacco-related health disparities: directions for future research. AB - Certain groups in the United States remain at high risk and suffer disproportionately from tobacco-related illness and death despite progress made in reducing tobacco use. To address gaps in research on tobacco-related disparities and develop a comprehensive agenda aimed at reducing such disparities, representatives from funding agencies, community-based organizations, and academic institutions convened at the National Conference on Tobacco and Health Disparities in 2002. Conference participants reviewed the current research, identified existing gaps, and prioritized scientific recommendations. Panel discussions were organized to address research areas affecting underserved and understudied populations. We report major research recommendations made by the conference participants in several scientific domains. These recommendations will ultimately help guide the field in reducing and eliminating tobacco-related disparities in the United States. PMID- 14759930 TI - The "global settlement" with the tobacco industry: 6 years later. AB - On June 20, 1997 a group of attorneys and health advocates proposed a "global settlement" of all public and private litigation against the tobacco industry. This agreement was controversial, and the subsequent implementing legislation was defeated. We sought to determine whether the global settlement represented a "missed opportunity" or a dead end. We compared the global settlement with subsequent laws, regulations, settlements, and judgments against the tobacco industry and found that other than Food and Drug Administration regulation of tobacco, tobacco control advocates have achieved many of the policies included in the global settlement and several beyond it. The policies that have been developed since 1997 have advanced tobacco control substantially, often beyond the provisions of the global settlement. PMID- 14759931 TI - Poor smokers, poor quitters, and cigarette tax regressivity. AB - The traditional view that excise taxes are regressive has been challenged. I document the history of the term regressive tax, show that traditional definitions have always found cigarette taxes to be regressive, and illustrate the implications of the greater price responsiveness observed among the poor. I explain the different definitions of tax burden: accounting, welfare-based willingness to pay, and welfare-based time inconsistent. Progressivity (equity across income groups) is sensitive to the way in which tax burden is assessed. Analysis of horizontal equity (fairness within a given income group) shows that cigarette taxes heavily burden poor smokers who do not quit, no matter how tax burden is assessed. PMID- 14759933 TI - Public health under attack: the American Stop Smoking Intervention Study (ASSIST) and the tobacco industry. AB - We describe the tobacco industry's response to the American Stop Smoking Intervention Study (ASSIST). Tobacco industry documents from the University of California, San Francisco/Legacy Tobacco Documents Library and industry Web sites were analyzed. LexisNexis and the Library of Congress's Thomas Web site were searched for legislative history. We found that the tobacco industry considered ASSIST a major threat because of its emphasis on policy and creation of local tobacco control infrastructures. The industry mobilized resources for a well coordinated attack on ASSIST. Although industry executives were sometimes frustrated in their efforts, they ultimately had a chilling effect on ASSIST. This evidence suggest that tobacco control advocates should expect a vigorous response from the tobacco industry to policy advocacy efforts, particularly at the local level. PMID- 14759932 TI - Reducing social disparities in tobacco use: a social-contextual model for reducing tobacco use among blue-collar workers. AB - In the United States in 1997, the smoking prevalence among blue-collar workers was nearly double that among white-collar workers, underscoring the need for new approaches to reduce social disparities in tobacco use. These inequalities reflect larger structural forces that shape the social context of workers' lives. Drawing from a range of social and behavioral theories and lessons from social epidemiology, we articulate a social-contextual model for understanding ways in which socioeconomic position, particularly occupation, influences smoking patterns. We present applications of this model to worksite-based smoking cessation interventions among blue-collar workers and provide empirical support for this model. We also propose avenues for future research guided by this model. PMID- 14759934 TI - Adult cigarette smoking prevalence: declining as expected (not as desired). AB - We compared observed smoking prevalence data for 1995-2002 with predictions derived from a previously published population dynamics model to determine whether the recent trend in smoking prevalence is consistent with the downward pattern we predicted. The observed data fit our projections closely (R 2 =.89). Consistent with the logic underlying the model, we conclude that adult smoking prevalence will continue to fall for the foreseeable future, although at a rate approximately half that of the decline experienced during the 1970s and 1980s. PMID- 14759935 TI - Sustained increased consumption of cigarettes, alcohol, and marijuana among Manhattan residents after september 11, 2001. AB - We compared reports of increased substance use in Manhattan 1 and 6 months after the September 11, 2001, terrorist attacks. Data from 2 random-digit-dial surveys conducted 1 and 6 months after September 11 showed that 30.8% and 27.3% of respondents, respectively, reported increased use of cigarettes, alcohol, or marijuana. These sustained increases in substance use following the September 11 terrorist attacks suggest potential long-term health consequences as a result of disasters. PMID- 14759936 TI - Confirming "truth": more evidence of a successful tobacco countermarketing campaign in Florida. AB - This study provides additional evidence that Florida's "truth" tobacco countermarketing campaign was successful in reducing smoking among Florida teens. Smoking rates were substantially lower among Florida teens between fall 2001 and spring 2002, whereas previous studies found that smoking rates were comparable before the launch of "truth." Florida teens had higher levels of "truth" campaign awareness and were more likely to agree with campaign-targeted beliefs; 2 of these beliefs were the only items associated with current smoking. PMID- 14759937 TI - Tobacco industry direct mail marketing and participation by New Jersey adults. AB - We examined adult participation in tobacco industry direct marketing: receipt of direct mail and use of coupons and brand reward programs. Participation was highest for direct mail; participation in all 3 forms differed by gender, age, and race/ethnicity; current smokers, Whites, and persons aged 25 to 64 years reported greater participation. Although tobacco industry direct marketing may influence smoking initiation, its potential to increase consumption and impede cessation is unquestionable. PMID- 14759938 TI - American Indian internet cigarette sales: another avenue for selling tobacco products. AB - A study conducted by the University of Minnesota found that cigarettes can be purchased on American Indian-owned Internet sites for about one fifth of the price at grocery stores, making this a more convenient, lower-priced, and appealing method of purchasing cigarettes. Researchers and educators are challenged to address this new marketing ploy and to discover ways to curb rising smoking rates in American Indian communities. PMID- 14759939 TI - Back to the future: Smoking in movies in 2002 compared with 1950 levels. AB - We reviewed smoking shown in a random sample of major motion pictures from 1950 through 2002. Smoking incidents declined from 10.7 incidents per hour in 1950 to a minimum of 4.9 in 1980-1982 but increased to 10.9 in 2002. Despite declining tobacco use and increasing public understanding of the dangers of smoking in the real world, smoking in movies has returned to levels observed in 1950, when smoking was nearly twice as prevalent in reality as it was in 2002. PMID- 14759940 TI - Project FLAVOR: 1-Year Outcomes of a Multicultural, School-Based Smoking Prevention Curriculum for Adolescents. AB - To evaluate a multicultural smoking prevention curriculum, 16 schools were randomized to receive the multicultural curriculum or a standard curriculum and program effects on 1-year smoking initiation among 1430 never smokers were assessed. Hispanic boys who received the multicultural curriculum were less likely to initiate smoking than were those who received the standard curriculum; effects were insignificant among other groups. The prevention effect among Hispanic boys is encouraging, but additional research is needed to improve prevention effects among other groups. PMID- 14759941 TI - Risk factors associated with problem use of prescription drugs. AB - We estimate the prevalence of and risk factors for the problem use of prescription drugs, overall and by therapeutic class. Applying logistic regression analysis to data from the National Household Survey on Drug Abuse, we found that nearly 1.3 million Americans aged 12 years and older experience problem use of prescription drugs signifying physiological dependence or heavy daily use. Those at greatest risk include older adults, females, those in poor/fair health, and daily alcohol drinkers. PMID- 14759942 TI - Working class matters: socioeconomic disadvantage, race/ethnicity, gender, and smoking in NHIS 2000. AB - OBJECTIVES: We sought to describe the burden of smoking on the US population, using diverse socioeconomic measures. METHODS: We analyzed data from the 2000 National Health Interview Survey. RESULTS: Overall, the prevalence of current smoking was greatest among persons in--and independently associated with--working class jobs, low educational level, and low income. Attempts to quit showed no socioeconomic gradient, while success in quitting was greatest among those with the most socioeconomic resources. These patterns held in most but not all race/ethnicity-gender groups. Finer resolution of smoking patterns was obtained using a relational UK occupational measure, compared to the skill-based measure commonly used in US studies. CONCLUSIONS: Reducing social disparities in smoking requires attention to the complexities of class along with race/ethnicity and gender. PMID- 14759943 TI - Effects of childhood socioeconomic circumstances on persistent smoking. AB - OBJECTIVES: We investigated whether socioeconomic circumstances at different life stages influence persistent smoking. METHODS: We followed a British birth cohort (all births between March 3 and 9, 1958) for 41 years to examine the influence of childhood and adulthood socioeconomic position on persistent smoking in adulthood (n = 6541). RESULTS: Persistent smoking (19% of participants, n = 1216) showed strong social gradients with both childhood and adulthood socioeconomic measures. Among men, the association with childhood socioeconomic circumstances was no longer significant after we adjusted for adulthood socioeconomic circumstances; however, among women, the adjusted odds of persistent smoking increased by 8% for each unit increase across a 16-point childhood score. CONCLUSIONS: Childhood socioeconomic circumstances predicted persistent smoking among women in our cohort, a finding that highlights the importance of influences on the development of persistent smoking across the life course. PMID- 14759944 TI - Town-level characteristics and smoking policy adoption in Massachusetts: are local restaurant smoking regulations fostering disparities in health protection? AB - OBJECTIVES: We identified and quantified differences in sociodemographic characteristics of communities relative to the strength of local restaurant smoking regulations in Massachusetts. METHODS: We examined the relationship between the strength of the 351 local restaurant smoking regulations in Massachusetts and a number of town-level characteristics, using a multinomial logistic regression model. RESULTS: Characteristics important to the adoption of stronger restaurant smoking regulations included higher education and per capita income, geographic region, voter support for a state cigarette tax initiative, board of health funding to promote clean indoor air policy making, and the presence of a bordering town with a strong regulation. CONCLUSIONS: The current pattern of smoke-free restaurant policy enactment fosters socioeconomic and geographic disparities in health protection, undermining an important national health goal. PMID- 14759945 TI - From adolescence to young adulthood: racial/ethnic disparities in smoking. AB - OBJECTIVES: We used data gathered from 6259 youths between the ages of 13 and 23 years to compare trends in smoking among 4 racial/ethnic groups. METHODS: We weighted trend data to represent baseline respondent characteristics and evaluated these data with linear contrasts derived from multiple regression analyses. RESULTS: Although African Americans exhibited higher initiation rates than Whites, they exhibited consistently lower rates of regular smoking than both Whites and Hispanics. This seeming anomaly was explained by African Americans' lower rates of transition to regular smoking and greater tendency to quit. Racial/ethnic disparities were accounted for by differences in pro-smoking influences. CONCLUSIONS: Reducing racial/ethnic disparities in smoking may require reducing differences in the psychosocial factors that encourage smoking. PMID- 14759946 TI - Acculturation and tobacco use among Chinese Americans. AB - OBJECTIVES: We examined the relationship between acculturation and tobacco use behaviors among Chinese Americans. METHODS: Using a Chinese-language instrument based on validated questions from several national surveys, we conducted in person, household-based interviews with 712 representative adults aged 18-74 years. RESULTS: Observed smoking prevalence was 29% for men and 4% for women. Predictors of smoking cessation included being 35 years and older and having a high level of tobacco-related knowledge. Acculturation was positively associated with a history of never smoking, as was being younger than 35 years and having a high level of tobacco-related knowledge. CONCLUSIONS: Acculturation was positively associated with never smoking among men but not with smoking cessation. However, knowledge of tobacco-related health risks was associated with both. Results indicate a need for language-specific educational interventions. PMID- 14759947 TI - Effects of tobacco smoke exposure on asthma prevalence and medical care use in North Carolina middle school children. AB - OBJECTIVES: We sought to determine the effects of exposure to environmental tobacco smoke (ETS) and childhood cigarette smoking on asthma symptoms among middle school children in North Carolina. METHODS: During 1999-2000, information was collected from a survey completed by the children. Outcomes of asthma symptom reporting were regressed on tobacco smoke exposures. RESULTS: Children who currently smoked or reported any exposure to ETS were at increased risk of reporting active asthma symptoms. Exposure to ETS and childhood cigarette smoking was responsible for 15% of the asthma cases observed in the study population and $1.34 million in excess medical expenditures. CONCLUSIONS: Even at low levels of exposure, childhood cigarette smoking and ETS are independently associated with asthmatic symptoms. PMID- 14759948 TI - Decrease in the prevalence of environmental tobacco smoke exposure in the home during the 1990s in families with children. AB - OBJECTIVES: This study explored correlates with and changes in the prevalence of environmental tobacco smoke (ETS) exposure of children in the home. METHODS: We used multiple logistic regression to explore ETS exposures as reported in the 1992 and 2000 National Health Interview Survey. RESULTS: ETS exposure in homes with children declined from 35.6% to 25.1% (P <.001) between 1992 and 2000, whereas smoking prevalence declined 26.5% to 23.3%. Home ETS exposures were more prevalent among non-Hispanic Whites than among African Americans (adjusted odds ratio [AOR] = 0.702; 95% confidence interval [CI] = 0.614, 0.802), Asian Americans (AOR = 0.534; 95% CI = 0.378, 0.754), and Hispanics (AOR = 0.388; 95% CI = 0.294, 0.389). Exposures declined across all groups, with greater gains in higher education and income groups. CONCLUSIONS: Home ETS exposure declined sharply between 1992 and 2000, more than would be predicted by the decline in adult smoking prevalence. PMID- 14759949 TI - Effect of local restaurant smoking regulations on environmental tobacco smoke exposure among youths. AB - OBJECTIVES: We examined the effect of local restaurant smoking regulations on restaurant environmental tobacco smoke exposure among youths. METHODS: We interviewed 3863 Massachusetts youths aged 12-17 years and ascertained how often they saw smokers in restaurants in their town. We assessed the effect of local restaurant smoking regulation strength on nonexposure to environmental tobacco smoke (seeing smokers never or only rarely). RESULTS: Compared with youths from towns with weak regulations, youths from towns with medium-strength regulations had 1.4 times the odds (odds ratio = 1.36; 95% confidence interval = 1.12, 1.65) and youths from towns with strong regulations had twice the odds (odds ratio = 2.03; 95% confidence interval = 1.64, 2.52) of reporting nonexposure. CONCLUSIONS: Strong local restaurant smoking regulations are associated with reduced environmental tobacco smoke exposure among youths PMID- 14759950 TI - Young adults: vulnerable new targets of tobacco marketing. AB - OBJECTIVES: We examined young adult smoking patterns and receptivity to cigarette advertising to assess vulnerability to tobacco marketing strategies. METHODS: We obtained data from a telephone survey of 12,072 Massachusetts adults. RESULTS: Smokers aged 18 to 30 years were more likely than older adults to smoke only occasionally and to consume fewer than 10 cigarettes per day. They also were more receptive to cigarette marketing and were more likely to be frequent patrons of bars and clubs. CONCLUSIONS: Many young adult smokers are in the initiation phase of smoking and are likely to undergo a transition to either nonsmoking or heavier smoking. If unimpeded by regulation, tobacco promotion in bars and clubs is likely to lead to increased adult smoking prevalence. PMID- 14759951 TI - Progression to established smoking among US youths. AB - OBJECTIVES: Our study presents national estimates of the proportion of youths in each of 7 stages of smoking and investigates the associations between risk/protective factors and progression to established smoking. METHODS: We analyzed data from the 1999 and 2000 National Youth Tobacco Surveys. RESULTS: In 1999 and 2000, 48.6% of US adolescents had at least experimented with tobacco, and 7.8% were established smokers. Important correlates of progression to established smoking included parental advice not to smoke, antismoking lessons in school, susceptibility to tobacco industry advertising and promotion, peer smoking, and exposure to smoking at home. CONCLUSIONS: Interventions to stop adolescent progression to established smoking should target susceptible never smokers and early experimenters as well as those in later stages of smoking. PMID- 14759952 TI - Neuroprotective effects of nicotinamide after experimental spinal cord injury. AB - OBJECTIVE: To investigate the ability of nicotinamide to protect against secondary damage in spinal cord tissue after an experimental injury. Trauma to the spinal cord induces a cascade of cellular events that lead to progressive tissue injury over time. Nicotinamide has been shown to affect many elements of this cascade, including excitatory amino acid release, inflammation, apoptosis, and cellular energy balance. METHODS: Male Long-Evans (n = 12) rats received an excitotoxic spinal cord injury by intraspinal injection of quisqualic acid (QUIS), a glutamate receptor agonist. A second set of rats (n = 4) received intraspinal saline as a sham injury. Thirty minutes after injury, animals that had QUIS injections received an intraperitoneal injection of either saline (control, n = 4) or nicotinamide (500 mg/kg, n = 8). Seven days postinjury, the spinal cords were removed, and serial sections were cut, mounted on slides, and stained. By using light microscopy, the extent of tissue damage was assessed at the epicenter of injury as well as sections up to 450- microm rostral and 450- microm caudal to the epicenter. RESULTS: Only those animals receiving QUIS injections showed damaged tissue. There was no significant difference in the amount of damage at the epicenter of injury between the saline- and nicotinamide treated animals. However, when comparing the total amounts of damage over the 975 microm length of cord examined, the rostro-caudal extent of injury was significantly reduced in the nicotinamide-treated animals compared with the saline-treated animals. CONCLUSIONS: Systemic nicotinamide serves to limit the rostro-caudal extent of cell death after experimental spinal cord injury. PMID- 14759953 TI - Xanthochromia is not pathognomonic for subarachnoid hemorrhage. AB - OBJECTIVE: To test the hypothesis that xanthochromia may be observed in traumatic lumbar puncture (LP). Xanthochromia, the yellow discoloration of cerebrospinal fluid (CSF) caused by hemoglobin catabolism, is classically thought to arise within several hours after subarachnoid hemorrhage (SAH). The presence of xanthochromic supernatant is often used to distinguish the elevated red blood cell (RBC) count observed in the CSF of SAH from the elevated RBC count observed after traumatic LP. METHODS: The authors developed a model of traumatic LP by adding whole blood to pigment-free CSF to obtain RBC concentrations of 0, 5000, 10000, 20000, 30000, and 40000 RBC/ microL. Supernatant from centrifuged samples was assessed for xanthochromia by spectrophotometry. Xanthochromia was considered present if the absorption followed a characteristic oxyhemoglobin curve with a maximal absorption greater than 0.023 at 415 nm. RESULTS: Samples with at least 30000 RBC/ microL demonstrated xanthochromia immediately. Samples with 20000 RBC/ microL demonstrated xanthochromia within one hour, and samples with 10000 RBC/ microL or less, within two hours. CONCLUSIONS: Cerebrospinal fluid xanthochromia may be observed within two hours after traumatic LP and sooner in samples with greater than 10000 RBC/ microL. Conversely, xanthochromia in traumatic LP with less than 5000 RBC warrants further investigation for SAH. When the CSF RBC count is elevated above 10000 RBC/ microL, or the time between sample acquisition and analysis is prolonged, the clinician should not rely on xanthochromia to confirm SAH. PMID- 14759954 TI - Utilization of the emergency department after self-inflicted injury. AB - OBJECTIVES: To compare emergency department (ED) utilization by individuals who present with self-inflicted injuries with utilization by control populations. Individuals with self-inflicted injuries commonly present to the ED, yet little research has been conducted on this population in this setting. METHODS: Individuals who had an ED presentation in 1995-1996 for a self-inflicted injury were tracked prospectively for three to four years of follow-up. This group was matched by age and gender to two groups: individuals who presented with asthma and individuals who presented with other complaints. Data on return visits to the ED were collected from an administrative database. Groups were compared on rates of return visits. RESULTS: There were 478 individuals randomly selected for each group. Individuals in the self-inflicted injury group had higher rates of return visits to the ED over the follow-up period: 232.7 visits per 100 person-years for the self-inflicted injury group, compared with 117.6 for the asthma group, and 83.0 for the "other" group (p < 0.001). The self-inflicted injury group had higher rates for many types of diagnoses: self-inflicted injuries, mental disorders, substance abuse, unintentional injuries, assault, headache pain, and other complaints (all p < 0.001). Patients with more than three repeat visits per year were more common in the self-inflicted injury group (20.1%) than the asthma or "other" groups (9.2% and 5.6%, respectively). CONCLUSIONS: Individuals who harm themselves are chronic users of the ED. The ED represents an opportune setting from which individuals can be directed to appropriate treatment programs. PMID- 14759955 TI - Efficacy of an educational Web site for educating physicians about bioterrorism. AB - OBJECTIVES: To determine whether a Web-based educational intervention improves emergency physicians' knowledge about bioterrorism and to survey physicians' knowledge and sources of information on bioterrorism. METHODS: Prospective randomized controlled trial using pre- and postintervention testing in hospitals. Participants were general and pediatric emergency medicine attending physicians, fellows, and fourth-year emergency medicine residents. All participants completed a pretest and attended a lecture on bioterrorism. Participants were then randomized to the Web intervention group that received continuous access to a bioterrorism educational Web site with weekly exposure to case scenarios of diseases due to biological agents, or the control group. Participants were retested after one and six months and surveyed to identify their sources of information and assess their knowledge. RESULTS: Sixty-three physicians completed the pretest. There was no difference in mean +/- standard deviation (SD) pretest scores between Web intervention (45% +/- 10%) and control (44% +/- 10%) groups (mean difference: 1.9%; 95% confidence interval [CI] = -6.7% to 2.9%). There was no significant difference between pre- and posttest scores among groups at one month (Web intervention 48% +/- 10% vs. control 45% +/- 10%; mean difference: 3.3%; 95% CI = -8.5% to 2.0%) and six months (Web intervention 51% +/- 8% vs. control 47% +/- 9%; mean difference: 3.8%; 95% CI = -8.8% to 1.2%). More than 60% of physicians cited media reports as their primary source of information on bioterrorism and believed that their knowledge of bioterrorism was limited after one month. CONCLUSIONS: Providing physicians information on bioterrorism through simulated cases and continuous access to an educational Web site does not increase knowledge of bioterrorism. Physicians are more likely to use media reports for their primary source of information. PMID- 14759956 TI - An evaluation of emergency medicine resident interaction time with faculty in different teaching venues. AB - OBJECTIVES: To measure actual emergency medicine (EM) resident interaction time with faculty and to investigate the potential to use direct observation as an assessment tool for the core competencies. By 2006 all EM residencies must implement resident assessment techniques of the six Accreditation Council for Graduate Medical Education core competencies. Emergency medicine educators recommend direct observation as the optimal evaluation tool for patient care, systems-based practice, interpersonal and communication skills, and professionalism. Continuous faculty presence in the emergency department (ED) is widely believed to facilitate direct observation as an assessment technique. METHODS: Observational study of EM resident-faculty interaction time during two hour periods. Study venues included two EDs, two trauma services, inpatient medicine, adult and pediatric intensive care units (ICUs), and a pediatric outpatient clinic. Using a priori definitions, the authors categorized faculty-EM resident interaction time as direct observation of patient care, indirect patient care, or non-patient care activities, and calculated total faculty interaction time. Subjects were blinded to the nature of the study, and data gathering was encrypted. RESULTS: Two hundred seventy observation periods of two hours each were conducted, sampling 32 EMR1, 33 EMR2-3, 41 EM, and 38 non-EM faculty. The mean total faculty interaction time ranged from a high of 30% (95% CI = 20% to 41%) in the pediatric ICU to a low of 10% (95% CI = 3% to 16%) on internal medicine wards. Overall, EM faculty interaction time was 20% (95% CI = 18% to 22%). Direct observation by faculty ranged from a high of 6% for EMR2-3s in the critical care areas of the ED (95% CI = 3% to 9%) to a low of 1% (95% CI = 0% to 2%) on internal medicine wards. Overall ED direct observation time was 3.6% (95% CI = 2.6% to 4.7%). Emergency department direct observation did not vary within EM resident training level or by ED site. Direct observation varied by treatment area within the EDs, with the critical care areas being substantially higher (6%) than the noncritical care areas (1%). CONCLUSIONS: Faculty direct observation time of EM residents was low in all training venues studied. Direct observation was the highest in ED critical care areas and lowest on medicine ward rotations. Emergency medicine faculty involved simultaneously in routine ED teaching, supervision, and patient care rarely performed direct observation, despite their continuous physical presence. This finding suggests that alternative strategies may be required to assess core competencies through direct observation in the ED. PMID- 14759957 TI - Evolution of an emergency department screening questionnaire for severe acute respiratory syndrome. AB - OBJECTIVES: To describe the screening tool that was used to screen for severe acute respiratory syndrome (SARS), the three revisions that were made, and the factors that led to these revisions. On March 13, 2003, on receiving notification of an outbreak of atypical pneumonia, nurses from the study emergency department (ED) started screening patients for the disease that became known as SARS. METHODS: The ED nurses started with a simple screening tool that was incorporated into triage. The screening tool was later revised into a questionnaire. An outdoor screening station was set up and patients were subsequently screened before triage. After the patients were screened, they were assigned to different risk areas, where triage and treatment were rendered. Two further revisions were made to the questionnaire. RESULTS: From March 13 to May 31, 11457 patients were screened. Version One of the screening questionnaire was used to screen 72 patients from March 13 to 17, Version Two screened 93 patients from March 18 to 21, Version Three screened 2909 patients from March 22 to April 8, and Version Four screened 8383 patients from April 9 to May 31. There was a significant (p < 0.05) downward trend in the proportion of admissions. Among those discharged from the ED, 0.28% reattended and were later confirmed to have SARS. CONCLUSIONS: The screening tool underwent three major revisions in response to new information. By keeping it relatively simple, user-friendly, and regularly updated, nurses were able to screen patients rapidly. Risk categorization ensured that no cross infection occurred among patients and that no one contracted SARS in the ED. PMID- 14759959 TI - Emergency department patient satisfaction: examining the role of acuity. AB - OBJECTIVES: To explore the relationships between patient acuity, perceived and actual throughput times, and emergency department (ED) patient satisfaction. The authors hypothesized that high-acuity patients would be the most satisfied with their throughput times, as well as the overall ED visit. The authors also expected overall ED satisfaction to be more strongly associated with perceived throughput times compared with actual throughput times, regardless of acuity. METHODS: This was a prospective survey of 1,865 ED patients at a large, inner city hospital during a one-month period. Data were collected on patient demographics, acuity of patient illness, actual waiting time for evaluation by a physician, and actual overall length of stay. Patient satisfaction with various throughput times (i.e., perceived throughput time) and overall ED visit was assessed by using a seven-point scale (1 = poor, 7 = excellent). Analysis of variance, analysis of covariance (ANCOVA), and correlations were conducted to explore the hypotheses. RESULTS: Patients with "emergent" acuity perceived their throughput times more favorably and were more satisfied with their overall ED visit compared with "urgent" and "routine" patients (all p < 0.01). Once the effects of perceived throughput time were controlled for by using an ANCOVA, acuity no longer predicted overall ED satisfaction. Correlations showed that overall ED satisfaction was more closely linked to perceived throughput times than to actual throughput times (average r = 0.62 vs. -0.12). CONCLUSIONS: "Emergent" patients are more satisfied than "urgent" and "routine" patients with their ED visits. "Emergent" patients perceived their throughput times more favorably than other patients, especially their wait for physician evaluation. Changing perceptions of throughput times may yield larger improvements in satisfaction than decreasing actual throughput times, regardless of patient acuity. PMID- 14759960 TI - Clinicopathological conference: fever, productive cough, and tachycardia in a 22 year-old Asian male. PMID- 14759961 TI - Should we screen for depression in the emergency department? PMID- 14759962 TI - Crash course in decision making. PMID- 14759963 TI - TRISS: does it get better than this? AB - The Trauma and Injury Severity Scoring (TRISS) system was developed in the 1980s to improve the prediction of patient outcomes following trauma through the use of physiological and anatomical criteria. The TRISS is used for a number of purposes, including quantifying the severity of injury of a patient population, calculating the probability of survival of patients for identification of cases for peer review, and comparing the death or survival rates of different populations/hospitals. Despite the advancements in trauma care, improved statistical techniques, and the identification of numerous limitations of TRISS, it continues to be the most commonly used tool for judging hospital performance and monitoring trauma death rates. This article critically evaluates the development, structure, and practical use of TRISS to determine its value in the current trauma environment. Limitations of TRISS are discussed and suggestions are made for the future development of trauma prediction tools. PMID- 14759964 TI - Advanced statistics: understanding medical record review (MRR) studies. AB - Medical record review (MRR) studies have been reported to make up 25% of all scientific studies published in emergency medical (EM) journals. However, unlike other study designs, there are no standards for reporting MRRs and very little literature on the methodology for conducting them. The purpose of this article is to provide the reader with methodological guidance regarding the strengths and weaknesses of these types of studies. PMID- 14759965 TI - Epidemiology of adult psychiatric visits to US emergency departments. AB - OBJECTIVES: To characterize psychiatric-related emergency department visits (PREDVs) among adults in the United States for the year 2000 and to analyze PREDV trends from 1992 to 2000. METHODS: Emergency department (ED) visit data from the National Hospital Ambulatory Medical Care Survey were used to estimate the number of PREDVs for adults aged 18 years and older. A PREDV was defined as any visit with a psychiatric discharge diagnosis (ICD N290- N312) or a suicide attempt (ICD E950-E959). RESULTS: Approximately 4.3 million PREDVs occurred in the United States in the year 2000, yielding an annual rate of 21 visits per 1000 adults. The PREDV rates increased 15% between 1992 and 2000. The PREDVs accounted for 5.4% of all ED visits. Substance abuse (27%), neuroses (26%), and psychoses (21%) were the most common conditions. African Americans had significantly higher visit rates (29/1000; 95% CI = 27/1000 to 31/1000) compared with whites (23/1000; 95% CI = 22/1000 to 25/1000). Persons with Medicaid (66/1000; 95% CI = 64/1000 to 68/1000) had double the rate of PREDVs than the uninsured (33/1000; 95% CI = 31/1000 to 35/1000) and almost eight times the rate of those privately insured (8/1000; 95% CI = 7/1000 to 10/1000). Patients with psychiatric diagnoses had a higher admission rate (22%) than those with nonpsychiatric diagnoses (15%). The uninsured were the least likely to be admitted for all major psychiatric conditions except suicide (p < 0.0001). CONCLUSIONS: Psychiatric-related ED visits represent a substantial and growing number of ED visits each year. Patient characteristics influence the likelihood of a PREDV. Further research is needed to better understand the role that hospital EDs play in the delivery of health care services to those with mental illness. PMID- 14759966 TI - Error identification, disclosure, and reporting: practice patterns of three emergency medicine provider types. AB - OBJECTIVES: To gather preliminary data on how the three major types of emergency medicine (EM) providers, physicians, nurses (RNs), and out-of-hospital personnel (EMTs), differ in error identification, disclosure, and reporting. METHODS: A convenience sample of emergency department (ED) providers completed a brief survey designed to evaluate error frequency, disclosure, and reporting practices as well as error-based discussion and educational activities. RESULTS: One hundred sixteen subjects participated: 41 EMTs (35%), 33 RNs (28%), and 42 physicians (36%). Forty-five percent of EMTs, 56% of RNs, and 21% of physicians identified no clinical errors during the preceding year. When errors were identified, physicians learned of them via dialogue with RNs (58%), patients (13%), pharmacy (35%), and attending physicians (35%). For known errors, all providers were equally unlikely to inform the team caring for the patient. Disclosure to patients was limited and varied by provider type (19% EMTs, 23% RNs, and 74% physicians). Disclosure education was rare, with 85% of eligible patients. Methods to obtain closer to 100% enrollment are desired. METHODS: During consecutive 15-day periods with the same 24 AAs, the daily ED census, hours of AA coverage, and enrollment in each of six studies were evaluated prospectively in the ED. Data are presented as means with 95% confidence intervals (CIs). RESULTS: There was no difference between the shared and split enrollment periods with respect to hours of AA coverage (30.3 vs. 30.7 hours/day; p = 0.7) or average daily ED census (133.7 vs. 141.8; p = 0.15). Overall, the percentages of ED patients recruited for study participation were not different depending on whether the split versus shared recruitment strategy was used (907 patients recruited out of 2005 ED patients (45.2%; 95% CI = 43.0 to 47.4) vs. 937 of 2127 (44.0%; 95% CI = 41.9 to 46.1). The 95% CI for the 1.2% difference was 1.8% to 4.2%. Patient enrollments in six individual studies were similar regardless of recruitment strategy. Following the 30-day trial, AAs were surveyed: 17 of 24 (71%) found the split strategy to be "more helpful in enrolling subjects," and 20 of 24 (83%) found split strategy helped them "keep better track" of patients. CONCLUSIONS: Study subject enrollment was not affected by the use of either the shared or split responsibility strategy for recruitment. Students generally preferred the split strategy because it was more helpful and easier to monitor. Therefore, this may be the best option for similar student oriented data collection programs. PMID- 14759968 TI - Proxy identification: a time-dependent analysis. AB - OBJECTIVES: To describe the availability of next-of-kin (NOK) for proxy consent over the 24-hour time period following presentation of major trauma patients to a Level I trauma center. METHODS: The study was conducted by using a prospective, observational study design. Consecutive patients meeting predefined major trauma criteria during a three-month study period were enrolled and followed until NOK were contacted, or up to 24 hours. Survival analysis was used to determine the probability of NOK contact during the 24-hour follow-up period. RESULTS: Three hundred fifty (92%) of 382 subjects had NOK contacted during the follow-up period. The probabilities (95% confidence intervals [CIs]) of NOK contact at the time of arrival (t = 0) and 2, 4, 6, and 24 hours postpresentation were 0.25 (95% CI = 0.21 to 0.30), 0.68 (95% CI = 0.63 to 0.72), 0.77 (95% CI = 0.73 to 0.82), 0.80 (95% CI = 0.76 to 0.84), and 0.87 (95% CI = 0.84 to 0.90), respectively. The median time to contact was 40 minutes (95% CI = 27 to 54 min). CONCLUSIONS: One third of the trauma patients in this study did not have NOK available two hours following arrival in the emergency department. For time-critical research protocols, the time to NOK contact can significantly impact the ability to ensure prompt enrollment and treatment in clinical trials. Further studies are needed to assess the generalizability and consistency of these data. PMID- 14759969 TI - Shedding new light on the "safe" club drug: methylenedioxymethamphetamine (ecstasy)-related fatalities. AB - OBJECTIVES: To report the pathology, toxicology, cause, and manner of death in 3,4-methylenedioxymethamphetamine (MDMA)-associated fatalities in the United States. Although use trends are increasing, data regarding the hazards of MDMA are limited. METHODS: The authors obtained fatality reports from participating medical examiners in the United States. Cases were identified as "drug-unrelated" when MDMA did not directly cause death (e.g., motor vehicle collision); deaths from drug toxicity were judged "drug-related." RESULTS: Thirty-eight (8%) of the surveyed medical examiners reported 102 deaths associated with MDMA use from 1999 to 2001. Ten percent of fatalities occurred in 1999 and 90% thereafter, representing a 400% relative increase. Victims tended to be young (mean age = 25), white (n = 87 [85%]), male (n = 70 [69%]), and otherwise healthy. Seventy one (70%) deaths were drug-related (DR) and 31 (30%) were drug-unrelated (DU). Twenty-four (35%) DR deaths had a mean delay of 6.7 hours (95% CI = 5.1 to 8.2) in activating emergency medical services. Fifty-five DR cases (81%) were found in asystole and pronounced dead at the scene. CONCLUSIONS: The MDMA-associated fatal events typically occur in young, otherwise healthy individuals. MDMA's impact on the public health and safety of young adults and teenagers needs further assessment. PMID- 14759970 TI - Needle thoracostomy: implications of computed tomography chest wall thickness. AB - OBJECTIVES: To determine the length of catheter required to perform a needle thoracostomy, as determined by chest wall thickness, to treat the majority of patients presenting to the emergency department (ED) with a potential tension pneumothorax. METHODS: A convenience sample of 111 computed tomography (CT) scans of the chest in trauma and medical resuscitation patients at a military Level 1 trauma center in San Antonio, Texas, was pooled, and the chest wall thickness was measured at the second intercostal space, midclavicular line, to the nearest 0.1 cm. RESULTS: The mean chest wall thickness in the patients studied averaged 4.24 cm (95% confidence interval [CI] = 3.97 to 4.52). Nearly one fourth (25) of the study patients had a chest wall thicker than 5 cm. Women, on average, have thicker chest walls than men (4.90 for women; 4.16 for men; p = 0.022). CONCLUSIONS: In this study, a catheter length of 5 cm would reliably penetrate the pleural space of only 75% of patients. A longer catheter should be considered, especially in women. PMID- 14759972 TI - Telltale heart skips a few beats: challenging conclusions of EMS customer satisfaction. PMID- 14759973 TI - Progress report: emergency medicine in southern Israel. PMID- 14759974 TI - Computer-aided detection of breast cancer: has promise outstripped performance? PMID- 14759975 TI - A model is a model: a decision analysis for rectal cancer. PMID- 14759976 TI - New Medicare bill targets discrepancies in fees for cancer drugs, outpatient services. PMID- 14759977 TI - Changes in Medicare payments for drug administration services. PMID- 14759978 TI - Stat bite: Medicare enrollment, 1966-2002. PMID- 14759979 TI - Politics and health: recent debate in Congress questions some government grants. PMID- 14759980 TI - NIH grants in question. PMID- 14759981 TI - America's first cancer center celebrates centennial. PMID- 14759982 TI - Can disease-free survival be a surrogate for overall survival for drug approvals? PMID- 14759984 TI - Workshop on partial breast irradiation: state of the art and the science, Bethesda, MD, December 8-10, 2002. AB - Breast conserving surgery followed by radiation therapy has been accepted as an alternative to mastectomy in the management of patients with early-stage breast cancer. Over the past decade there has been increasing interest in a variety of radiation techniques designed to treat only the portion of the breast deemed to be at high risk for local recurrence (partial-breast irradiation [PBI]) and to shorten the duration of treatment (accelerated partial-breast irradiation [APBI]). To consider issues regarding the equivalency of the various radiation therapy approaches and to address future needs for research, quality assurance, and training, the National Cancer Institute, Division of Cancer Treatment and Diagnosis, Radiation Research Program, hosted a Workshop on PBI in December 2002. Although 5- to 7-year outcome data on patients treated with PBI and APBI are now becoming available, many issues remain unresolved, including clinical and pathologic selection criteria, radiation dose and fractionation and how they relate to the standard fractionation for whole breast irradiation, appropriate target volume, local control within the untreated ipsilateral breast tissue, and overall survival. This Workshop report defines the issues in relation to PBI and APBI, recommends parameters for consideration in clinical trials and for reporting of results, serves to enhance dialogue among the advocates of the various radiation techniques, and emphasizes the importance of education and training in regard to results of PBI and APBI as they become emerging clinical treatments. PMID- 14759985 TI - Changes in breast cancer detection and mammography recall rates after the introduction of a computer-aided detection system. AB - BACKGROUND: Computer-aided mammography is rapidly gaining clinical acceptance, but few data demonstrate its actual benefit in the clinical environment. We assessed changes in mammography recall and cancer detection rates after the introduction of a computer-aided detection system into a clinical radiology practice in an academic setting. METHODS: We used verified practice- and outcome related databases to compute recall rates and cancer detection rates for 24 Mammography Quality Standards Act-certified academic radiologists in our practice who interpreted 115,571 screening mammograms with (n = 59,139) or without (n = 56,432) the use of a computer-aided detection system. All statistical tests were two-sided. RESULTS: For the entire group of 24 radiologists, recall rates were similar for mammograms interpreted without and with computer-aided detection (11.39% versus 11.40%; percent difference = 0.09, 95% confidence interval [CI] = 11 to 11; P =.96) as were the breast cancer detection rates for mammograms interpreted without and with computer-aided detection (3.49% versus 3.55% per 1000 screening examinations; percent difference = 1.7, 95% CI = -11 to 19; P =.68). For the seven high-volume radiologists (i.e., those who interpreted more than 8000 screening mammograms each over a 3-year period), the recall rates were similar for mammograms interpreted without and with computer-aided detection (11.62% versus 11.05%; percent difference = -4.9, 95% CI = -21 to 4; P =.16), as were the breast cancer detection rates for mammograms interpreted without and with computer-aided detection (3.61% versus 3.49% per 1000 screening examinations; percent difference = -3.2, 95% CI = -15 to 9; P =.54). CONCLUSION: The introduction of computer-aided detection into this practice was not associated with statistically significant changes in recall and breast cancer detection rates, both for the entire group of radiologists and for the subset of radiologists who interpreted high volumes of mammograms. PMID- 14759986 TI - Impact of preoperative staging and chemoradiation versus postoperative chemoradiation on outcome in patients with rectal cancer: a decision analysis. AB - BACKGROUND: Although radical resection and postoperative chemoradiation have been the standard therapy for patients with rectal cancer, preoperative staging by local imaging and chemoradiation are widely used. We used a decision analysis to compare the two strategies for rectal cancer management. METHODS: We developed a decision model to compare survival outcomes after postoperative chemoradiation versus preoperative staging and chemoradiation in patients aged 70 years with resectable rectal cancer. In the postoperative chemoradiation strategy, patients undergo radical resection and receive postoperative chemoradiation. In the preoperative staging and chemoradiation strategy, patients with locally advanced cancer receive preoperative chemoradiation and radical resection, whereas those with amenable localized tumors undergo local excision. The cohorts of patients were entered into a Markov model incorporating age-adjusted and disease-specific mortality. Outcomes were evaluated by modeling 5-year disease-specific survival for preoperative chemoradiation as less than, equal to, or greater than that of postoperative chemoradiation. Base-case probabilities were derived from published data; the Surveillance, Epidemiology, and End Results (SEER) Program database; and U.S. Life Tables. One-way and two-way sensitivity analyses were performed. The outcome measures were life expectancy and quality-adjusted life expectancy. RESULTS: Life expectancy and quality-adjusted life expectancy were 9.72 and 8.72 years, respectively, in the postoperative chemoradiation strategy. In the preoperative staging and chemoradiation strategy, life expectancy was 9.36, 9.72, and 10.09 years and quality-adjusted life expectancy was 8.71, 9.04, and 9.37 years when 5-year disease-specific survival was less than, equal to, or greater than that of postoperative chemoradiation, respectively. The decision model was sensitive to differences in the long-term toxicity of pre- and postoperative chemoradiation. When the 5-year disease-specific survival for patients after pre- or postoperative chemoradiation was equal, the decision model was sensitive to surgical mortality and to the probability of residual lymph node disease after local excision. CONCLUSION: If efficacy and toxicity after preoperative chemoradiation are equal to or better than that after postoperative chemoradiation in patients with locally advanced rectal cancer, then preoperative staging to select patients appropriate for preoperative chemoradiation is beneficial. PMID- 14759987 TI - Mobilization of dendritic cell precursors into the circulation by administration of MIP-1alpha in mice. AB - BACKGROUND: Dendritic cells (DCs) play a central role in immune responses and may be useful adjuvants for tumor vaccine therapy. We previously reported that F4/80( )B220(-)CD11c(+) DC precursors expressing the CC chemokine receptors CCR1 and CCR5 are mobilized rapidly into the circulation in mice injected with Propionibacterium acnes and are recruited into inflammatory tissue by macrophage inflammatory protein 1alpha (MIP-1alpha), which binds to CCR1 and CCR5. Here we investigate the mechanisms of DC precursor mobilization and the antitumor effect of these cells in mice. METHODS: Numbers of DC precursors in peripheral blood were determined in P. acnes-treated mice (groups of 10 C57BL/B6 [B6] wild-type mice, CCR1(-/-) mice, CCR5(-/-) mice, and B6 mice treated with antibody to MIP 1alpha or control antibody) and in B6 mice injected with recombinant MIP-1alpha. MIP-1alpha-mobilized DC precursors matured by treatment with granulocyte macrophage colony-stimulating factor, interleukin 4, and tumor necrosis factor alpha and pulsed with B16 melanoma lysates were assayed for their ability to confer protective immunity against tumor challenge in vivo and to induce cytotoxic T lymphocytes against B16 tumor cells in vitro. RESULTS: The recruitment of DC precursors into the circulation by P. acnes administration was higher in B6 mice (12.6%, 95% confidence interval [CI] = 9.1% to 16.1%) than in CCR1(-/-) (9.0%, 95% CI = 7.5% to 10.5%), CCR5(-/-) (6.3%, 95% CI = 5.2% to 7.3%), or anti-MIP-1alpha antibody-treated (6.6%, 95% CI = 5.7% to 7.5%) mice. Injection of MIP-1alpha also mobilized DC precursors into the circulation (13.1%, 95% CI = 10.8% to 15.6%). Matured MIP-1alpha-mobilized-DC precursors pulsed with B16 tumor lysates elicited B16-specific antitumor immunity in vitro and in vivo. CONCLUSIONS: MIP-1alpha and its receptors are important in recruiting DC precursors into the circulation. DC precursors mobilized rapidly by MIP-1alpha may provide sufficient useful DC precursors for DC-based vaccination in cancer treatment. PMID- 14759988 TI - Characterization of new estrogen receptor destabilizing compounds: effects on estrogen-sensitive and tamoxifen-resistant breast cancer. AB - BACKGROUND: Antiestrogens of the selective estrogen receptor modulator (SERM) type, such as tamoxifen, have two major limitations: their mixed agonist and antagonist profile and the development of tumor resistance. We characterized two new pure antiestrogens-ZK-703 and ZK-253-that belong to the class of specific estrogen receptor destabilizers (SERDs), which includes fulvestrant, and compared their activity with that of fulvestrant and tamoxifen. METHODS: Effects of antiestrogens on the growth of estrogen-dependent breast tumors in vivo were determined using several mouse xenograft models (including the tamoxifen sensitive tumors MCF7, T47D, and MV3366 and the tamoxifen-resistant tumors ZR75-1 and MCF7/TAM) and chemically induced (nitrosomethyl urea [NMU] and dimethylbenzanthracene [DMBA]) rat breast cancer models (groups of 10 animals). We determined the initial response and effects on hormone receptor levels and the time to relapse after treatment (i.e., time to reach a predetermined tumor size threshold). Estrogen receptor (ER) levels were determined by immunoassay. RESULTS: ZK-703 (administered subcutaneously) and ZK-253 (administered orally) were more effective than tamoxifen or fulvestrant at inhibiting the growth of ER positive breast cancer in all xenograft models. For example, MCF7 tumors relapsed (i.e., reached the size threshold) in 10 weeks in mice treated with tamoxifen but in 30 weeks in mice treated with ZK-703. ZK-703 and ZK-253 also prevented further tumor progression in tamoxifen-resistant breast cancer models to a similar extent (more than 30 weeks in mice with ZR75-1 and MCF7/TAM tumors). In the chemically induced rat breast cancer models, orally administered ZK-703 and ZK-253 caused a nearly complete (>80%) inhibition of tumor growth. ER levels were dramatically reduced in MCF7 tumors after 5 weeks of ZK-703 treatment compared with ER levels in vehicle-treated tumors; by contrast, ER levels in tamoxifen-treated tumors were higher than those in control tumors. CONCLUSION: ZK-703 and ZK-253 are potent, long-term inhibitors of growth in both tamoxifen-sensitive and tamoxifen resistant breast cancer models. PMID- 14759989 TI - Risk factors for breast cancer according to estrogen and progesterone receptor status. AB - BACKGROUND: Evaluations of epidemiologic risk factors in relation to breast cancer classified jointly by estrogen receptor (ER) and progesterone receptor (PR) status have been inconsistent. To address this issue, we conducted a prospective evaluation of risk factors for breast cancer classified according to receptor status. METHODS: During 1 029 414 person-years of follow-up of 66 145 women participating in the Nurses' Health Study from 1980 through 2000, we identified 2096 incident cases of breast cancer for which information on ER/PR status was available: 1281 were ER+/PR+, 318 were ER+/PR-, 80 were ER-/PR+, and 417 were ER-/PR-. We fit a log-incidence model of breast cancer and used polychotomous logistic regression to compare coefficients for breast cancer risk factors in patients with different ER/PR status. To test for differences in risk factor odds ratios based on marginal ER/PR categories, we evaluated ER status controlling for PR status and vice versa. The predictive ability of our log incidence model to discriminate between women who would develop ER+/PR+ breast cancer and those who would not (and similarly for ER-/PR- breast cancer) was evaluated by using receiver operator characteristic curve analysis. All statistical tests were two-sided. RESULTS: We observed statistically significant heterogeneity among the four ER/PR categories for some risk factors (age, menopausal status, body mass index [BMI] after menopause, the one-time adverse effect of first pregnancy, and past use of postmenopausal hormones) but not for others (benign breast disease, family history of breast cancer, alcohol use, and height). The one-time adverse association of first pregnancy with incidence was present for PR- but not for PR+ tumors after controlling for ER status (P =.007). However, the association of BMI after menopause with incidence was present for PR+ but not PR- tumors (P =.005). Statistically significant differences in the incidence of ER+ and ER- tumors were seen with age, both before and after menopause (P =.003), and with past use of postmenopausal hormones (P =.01). Area under the receiver operator characteristic curve, adjusted for age, was 0.64 (95% confidence interval [CI] = 0.63 to 0.66) for ER+/PR+ tumors and 0.61 (95% CI = 0.58 to 0.64) for ER-/PR- tumors. CONCLUSIONS: Incidence rates and risk factors for breast cancer differ according to ER and PR status. Thus, to accurately estimate breast cancer risk, breast cancer cases should be divided according to the ER and PR status of the tumor. PMID- 14759990 TI - Dietary glycemic load and risk of colorectal cancer in the Women's Health Study. AB - Although diet is believed to influence colorectal cancer risk, the long-term effects of a diet with a high glycemic load are unclear. The growing recognition that colorectal cancer may be promoted by hyperinsulinemia and insulin resistance suggests that a diet inducing high blood glucose levels and an elevated insulin response may contribute to a metabolic environment conducive to tumor growth. We prospectively followed a cohort of 38 451 women for an average of 7.9 years and identified 174 with incident colorectal cancer. We used baseline dietary intake measurements, assessed with a semiquantitative food-frequency questionnaire, to examine the associations of dietary glycemic load, overall dietary glycemic index, carbohydrate, fiber, nonfiber carbohydrate, sucrose, and fructose with the subsequent development of colorectal cancer. Cox proportional hazards models were used to estimate relative risks (RRs). Dietary glycemic load was statistically significantly associated with an increased risk of colorectal cancer (adjusted RR = 2.85, 95% confidence interval [CI] = 1.40 to 5.80, comparing extreme quintiles of dietary glycemic load; P(trend) =.004) and was associated, although not statistically significantly, with overall glycemic index (corresponding RR = 1.71, 95% CI = 0.98 to 2.98; P(trend) =.04). Total carbohydrate (adjusted RR = 2.41, 95% CI = 1.10 to 5.27, comparing extreme quintiles of carbohydrate; P(trend) =.02), nonfiber carbohydrate (corresponding RR = 2.60, 95% CI = 1.22 to 5.54; P(trend) =.02), and fructose (corresponding RR = 2.09, 95% CI = 1.13 to 3.87; P(trend) =.08) were also statistically significantly associated with increased risk. Thus, our data indicate that a diet with a high dietary glycemic load may increase the risk of colorectal cancer in women. PMID- 14759991 TI - Re: Integrin beta3 Leu33Pro homozygosity and risk of cancer. PMID- 14759992 TI - Re: Radiation therapy in the treatment of Hodgkin's disease--do you see what I see? PMID- 14759997 TI - Re: Zinc supplement use and risk of prostate cancer. PMID- 14759996 TI - Re: International variation in screening mammography interpretations in community based programs. PMID- 14760000 TI - "Morally reprehensible". PMID- 14760001 TI - Is there a role for cloning in human reproduction? PMID- 14760002 TI - Trainee page: Careers in andrology. PMID- 14760003 TI - Artificial vagina for rabbits. PMID- 14760004 TI - Melatonin administration alters semen quality in normal men. PMID- 14760005 TI - Genetic analysis of Kit ligand functions during mouse spermatogenesis. PMID- 14760006 TI - Zyxin, axin, and Wiskott-Aldrich syndrome protein are adaptors that link the cadherin/catenin protein complex to the cytoskeleton at adherens junctions in the seminiferous epithelium of the rat testis. AB - During spermatogenesis, the movement of germ cells across the seminiferous epithelium is associated with extensive junction restructuring. Yet the underlying mechanism (or mechanisms) that regulates these events is largely unknown. If the molecular architecture of the cell-cell actin-based adherens junction (AJ), such as ectoplasmic specialization (ES) and tubulobulbar complex- two testis-specific AJ types, is known, many functional mechanistic studies can be designed. We thus undertook an investigation to study 3 adaptors in the seminiferous epithelium: zyxin, axin, and Wiskott-Aldrich syndrome protein (WASP). All 3 adaptors were shown to be products of Sertoli and germ cells. Zyxin was shown to be a stage-specific protein that was most prominent during stages V VII and restricted mostly to pachytene spermatocytes, but it could also be detected at the site of basal and apical ectoplasmic specialization (ES). Zyxin, axin, and WASP were shown to be structurally linked to the N-cadherin/beta catenin/alpha-actinin/actin complex but not to the nectin-3/afadin or the beta 1 integrin-mediated protein complexes. Interestingly, zyxin, axin, and WASP are also structurally linked to vimentin (an intermediate filament protein) and alpha tubulin (the subunit of a microtubule), which suggests that they have a role (or roles) in the regulation of the dynamics of the desmosome-like junction and microtubule. These results illustrate that zyxin, axin, and WASP are adaptors in both AJs and intermediate filament-based desmosome-like junctions. This raises the possibility that classic cadherins are also associated with vimentin-based intermediate filaments via these adaptors in the testis. While virtually no N cadherin was found to associate with vimentin in the seminiferous tubules, it did associate with vimentin when testis lysates were used. Interestingly, about 5% of the E-cadherin associated with vimentin in isolated seminiferous tubules, and about 50% of the E-cadherin in the testis used vimentin as its attachment site. These data suggest that cadherins in the testis, unlike those in other epithelia, use different attachment sites to anchor the cadherin/catenin complex to the cytoskeleton. The levels of zyxin, axin, and WASP were also assessed during AF 2364-mediated AJ disruption of the testis, which illustrated a time-dependent protein reduction that was similar to the trends observed in nectin-3 and afadin but was the opposite of those observed for N-cadherin and beta-catenin, which were induced. Collectively, these results illustrate that while these adaptors are structurally associated with the cadherin/catenin complex in the testis, they are regulated differently. PMID- 14760007 TI - Effects of putative epididymal osmolytes on sperm volume regulation of fertile and infertile c-ros transgenic Mice. AB - Volume regulation by spermatozoa has been demonstrated to be crucial in both mice and men for transport in the female tract. In order to determine the nature of osmolytes used by spermatozoa, they were released from the cauda epididymis of fertile c-ros heterozygous mice into incubation medium of uterine osmolality (representing an osmotic challenge), containing increasing concentrations of compounds that are major epididymal fluid components and known osmolytes in somatic cells. This should nullify the concentration gradients for osmolytes that mediate volume regulation, prevent osmolyte efflux, and lead to swelling. Of the osmolytes tested, K(+) caused the most rapid and extensive volume increases; glutamate, taurine, L-carnitine, and myo-inositol also were effective, but glycerophosphocholine was not. Such effects were not observed in cauda sperm from the infertile knockout mice, demonstrating a defect in normal volume regulation. K(+) concentrations in cauda epididymal fluid were 21 mM higher in the knockout than the heterozygous mice, but no differences were found in caudal fluid glutamate, carnitine, or myo-inositol. The carnitine content of cauda sperm from knockout males was not different from that of fertile males, but lower amounts of glutamate and inositol were found that could explain the poor volume regulation. In heterozygous mice, cauda but not caput sperm responded to the K(+) channel blocker quinine by swelling, demonstrating development of volume regulation during epididymal transit, whereas knockout cauda sperm showed no response, as with the osmolytes. Major epididymal secretions could serve as osmolytes in murine spermatozoa for volume regulation in response to physiological osmotic challenge in the normal fertile mice; the reduced sperm content of inositol and glutamate in the c-ros knockout mice might reflect maturational abnormalities in volume regulation. PMID- 14760008 TI - Cryopreservation of ram semen facilitates sperm DNA damage: relationship between sperm andrological parameters and the sperm chromatin structure assay. AB - We hypothesized that cryopreservation and incubation in conditions that mimic the female genital tract following insemination increases the susceptibility of ram sperm DNA to denaturation. Ram sperm samples (n = 12) underwent the sperm chromatin structure assay (SCSA) and semen quality tests, including motility parameters, viability, and chlortetracycline fluorescence (CTC) patterns. We also assessed correlations between SCSA variables and semen quality parameters. Analyses were performed for both fresh and cryopreserved samples at 0, 3, and 20 hours of incubation in synthetic oviductal fluid (SOF; 39 degrees C, 5% CO(2)). The SCSA variables, mean alpha t (X alpha(t)) and standard deviation of alpha t (SD alpha(t)), were higher because of cryopreservation (P <.05, P <.001, respectively) after 20 hours in SOF. For both fresh and frozen spermatozoa, SCSA values (X alpha(t), SD alpha(t), and the percentage of cells outside the main population of alpha(t) [%COMP alpha(t)]) increased during incubation in SOF. Motility was negatively correlated with both SD alpha(t) and %COMP alpha(t), ranging from -0.39 (P <.01) to -0.59 (P <.001) for both fresh and cryopreserved semen; viability also was negatively correlated with X alpha(t), SD alpha(t), or %COMP alpha(t) (-0.36; P <.05, -.40 and -.46; P <.01, respectively) in fresh semen. The %COMP alpha(t) was positively correlated to the percentage of CTC pattern AR (P <.001) and negatively correlated to the percentages of patterns F and B (-0.33 to -0.60, P <.05 to P <.001). Variation among ejaculates within ram was observed (P <.01). Cryopreservation clearly facilitates DNA damage in physiological conditions. The low to moderate correlations between SCSA variables and classical semen quality parameters indicate that the SCSA provides additional information to standard tests for evaluating ram sperm quality. PMID- 14760009 TI - Interaction of PDC-109, the major secretory protein from bull seminal vesicles, with bovine sperm membrane Ca2+-ATPase. AB - PDC-109 is the prevalent secretory protein from bovine seminal vesicles that binds to the midpiece of sperm once they pass the ampulla of the vas deferens during emission. Thereby, the protein changes biophysical membrane properties, eventually resulting in increased sperm motility. To elucidate the underlying biochemical mechanism, we have studied the ion-pumping activity (Ca(2+)-ATPase) in membrane preparations of bovine spermatozoa following in vitro incubation with the protein and analyzed whether PDC-109 influences sperm motility. PDC-109 was purified to homogeneity from bull seminal vesicle extracts using a newly described method. The effect of PDC-109 on sperm motility was analyzed using the CASA-method. These experiments clearly demonstrated that PDC-109 significantly increases sperm motility. Calcium-pumping mechanisms were analyzed by monitoring the effect of PDC-109 on various parameters of enzyme activity of Ca(2+)-ATPase in epididymal sperm plasma membranes and were compared with Ca(2+)-ATPase activities from other organs and from epididymal sperm of different species, respectively. Specificity studies were performed using different Ca(2+) antagonists. Enzyme activities of both Mg(2+)-dependent and Mg(2+)-independent Ca(2+)-ATPases increased in a dose-dependent manner following the addition of the PDC-109 (range 5-20 microg). Preincubation of PDC-109 at temperatures above 37 degrees C and pHs ranging from below 6.5 and above 8.5 led to the loss of the stimulatory effect. An analysis of enzyme kinetics pointed to irreversible, cooperative interaction of PDC-109 with the enzyme. The effect was organ specific, that is, restricted to sperm ATPases, but it was not species-specific, as it could be elicited also in rat sperm. PMID- 14760010 TI - Role of nitric oxide concentrations on human sperm motility. AB - Nitric oxide (NO) is a free radical generated from the oxidation of L-arginine to L-citrulline by 3 isoforms of reduced nicotinamide adenine dinucleotide phosphate (NADPH)-dependent NO synthases. Several data suggest a relevant role in sperm cell pathophysiology, but any conclusive data on its role in spermatozoa motility are still lacking. In the present study, we have correlated NO concentration in semen and kinetic features of sperm cells from normozoospermic fertile donors and infertile patients affected by idiopathic asthenozoospermia. Normozoospermic fertile men exhibited NO concentrations that were significantly lower than those of asthenozoospermic infertile men. A significant linear negative correlation was evident between NO concentration and percentage of total sperm motility. A further significant linear negative correlation was found between NO concentration and spermatozoa kinetic characteristics determined by a computerized analysis (curvilinear and straight progressive velocity). These data suggest that the overproduction of this free radical and the consequent excessive exposure to oxidative conditions have a potential pathogenetic implication in the reduction of sperm motility. The positive role played by NO in spermatozoa capacitation leads us to speculate that such paradoxical involvement in both pathologic and physiologic processes depends on the alternative redox state and relative level of NO. PMID- 14760011 TI - Inflammatory cytokine concentrations are elevated in seminal plasma of men with spinal cord injuries. AB - The semen of most men with spinal cord injury (SCI) contains sperm with abnormally low motility. Studies suggest that the seminal plasma is the source of this condition. The seminal plasma of men with SCI contains an abnormally high number of white blood cells (WBC), specifically, activated T cells. It is known that activated T cells secrete cytokines and elevated concentrations of cytokines can be harmful to sperm. It is not known if the seminal plasma of men with SCI contains elevated concentrations of cytokines. The purpose of this study was to determine if the seminal plasma of men with SCI contained elevated concentrations of cytokines. Using the method of enzyme-linked immunosorbant assay (ELISA), ten cytokines were measured in the seminal plasma of men with SCI as well as healthy non-SCI control subjects. The cytokines of interest were grouped according to Th1 effector functions: interleukin 1 beta, interleukin 2, interleukin 12, tumor necrosis factor alpha, tumor necrosis factor beta, interferon gamma (IL1 beta, IL2, IL12, TNF alpha, TNF beta, INF gamma, respectively) and Th2 effector functions: interleukin 4, interleukin 6, interleukin 10, transforming growth factor beta 1 (IL4, IL6, IL10, TGF beta 1, respectively). The results showed a predominance of Th1 versus Th2 cytokine production in the seminal plasma of men with SCI compared with that of control subjects. This finding suggests an immunologic basis for infertility as a possible avenue of investigation in these men. PMID- 14760012 TI - A flow cytometric method for rapid determination of sperm concentration and viability in mammalian and avian semen. AB - A new flow cytometric method has been developed to rapidly determine sperm concentration and viability in semen from bulls and boars. Sperm viability was determined on the basis of staining with SYBR-14 and propidium iodide (PI), and this allowed detection of live (membrane-intact) sperm, dying (moribund) sperm, as well as dead cells. Fluorescent microspheres (beads) were used to determine sperm concentration. The use of SYBR-14 at 50 nM and PI at 12 micro M in combination with the FACSCount diluent in the counting tubes resulted in a uniform staining after 2.5-5 minutes at room temperature. Reagent staining was reproducible enough to allow subsequent semiautomated analysis of data using Attractors software. In experiment 1, this method was used to analyze semen from boars, rams, rats, rabbits, humans, and turkeys. In experiment 2, Attractors analysis was performed by the FACSCount AF flow cytometer, and sperm concentration determination with this system was compared with results obtained by a spectrophotometer and an electronic cell counter, which is routinely used by bull artificial insemination centers. When compared to microscopic counting in a hemocytometer, the FACSCount AF flow cytometer was two and four times more accurate than the spectrophotometer and the electronic cell counter, respectively. In addition, the FACSCount AF flow cytometer determined both sperm concentration (coefficient of variation 3.3%) and sperm viability (coefficient of variation 0.7%) with high precision. PMID- 14760013 TI - Immunolocalization and regulation of cystic fibrosis transmembrane conductance regulator in the adult rat epididymis. AB - Cystic fibrosis is the most common serious autosomal recessive condition in whites, and more than 95% of men with cystic fibrosis are infertile. The cystic fibrosis transmembrane conductance regulator (CFTR), a cyclic adenosine monophosphate (cAMP)-regulated chloride channel, has been localized in the efferent ducts; however, to our knowledge, its expression and regulation in the epididymis by testicular factors have not been examined. In the present study, these parameters were examined immunocytochemically by the light microscope with an anti-CFTR antibody in Bouin-fixed, paraffin-embedded control adult rat epididymides and both orchidectomized adult rats with or without testosterone supplementation and efferent duct-ligated rats sacrificed at different time points. In control animals, a thick dense band of immunoperoxidase reaction product was visualized over the apical plasma membrane of the principal cells but not their microvilli. The apical band was prominent only in the corpus and cauda regions. While there was no CFTR expression in basal cells, clear cells of the corpus and cauda regions showed a weak-to-moderate band of apical plasma membrane staining. An examination of orchidectomized, orchidectomized and testosterone, and efferent duct-ligated rats revealed that CFTR was no longer expressed as an intense band on the apical plasma membrane of the principal cells of the corpus and cauda regions. However, under these conditions, an intense apical/supranuclear reaction was noted in the form of small vesicular structures. Clear cells were unaffected by the different experimental treatments. Together, these data indicate that CFTR is expressed in a cell- and region-specific manner and that, while its synthesis in principal cells is not under the control of testicular factors, targeting to the apical plasma membrane is regulated by a testicular luminal factor. PMID- 14760014 TI - Limited processing of pro-matrix metalloprotease-2 (gelatinase A) overexpressed by transfection in PC-3 human prostate tumor cells: association with restricted cell surface localization of membrane-type matrix metalloproteinase-1. AB - The expression and activation of matrix metalloproteinases (MMPs) by tumor cells is correlated with progression to invasive and metastatic status. The purpose of this study was to examine the role of increased MMP-2 (gelatinase A) expression in prostate cancer progression utilizing human prostate PC-3 cancer cells that overexpress MMP-2 using gene transfection. PC-3 cells were transfected with pCR-3 vector only and pCR-3 MMP-2 plasmids employing the LipofectAMINE method, and stable transfectants were selected with G418. The expression of MMP-2, tissue inhibitor of metalloproteinase-2 (TIMP-2), and membrane-type MMP 1 (MT1-MMP) in PC-3 parental and transfected cells under serum-free conditions was determined by zymography, immunoblotting, immunofluorescent microscopy, Northern blotting, and/or reverse transcriptase-polymerase chain reaction (RT-PCR). MMP-2 transfected cells produced primarily the proenzyme form of MMP-2; the parental and vector control transfected PC-3 cells did not express any MMP-2 that was detectable by the methods we employed. Treatment of PC-3 MMP-2 transfected cells with Concanavalin A (Con A), in contrast to HT-1080 cells, processed only a small amount of the secreted 72-kd proenzyme to a 62-kd intermediate and a cell associated 59-kd active form. The low level of secreted pro-MMP-2 processing induced by Con A was inhibited by serine protease inhibitors and was unaffected by cyclic adenosine monophosphate (cAMP). Immunoblotting showed that these cells produced abundant TIMP-2 and lower amounts of MT1-MMP in comparison with Con A responding HT-1080 cells. HT-1080 cells respond to Con A by translocating MT1-MMP from intracellular localization sites to the plasma membrane, an effect not observed in PC-3 cells. The molecular basis for the low level of processing of pro-MMP-2 by PC-3 cells may be due to an overabundance of TIMP-2 and/or a low level of cell surface active MT1-MMP. PMID- 14760015 TI - Involvement of A1 adenosine receptors in the acquisition of fertilizing capacity. AB - Ejaculated mammalian spermatozoa acquire competence to fertilize oocytes by a two step process: capacitation followed by acrosome reaction. The biochemical and biophysical modifications occurring in vivo in the female reproductive tract can be reproduced in vitro, and previous studies have suggested a capacitative role for adenosine A(1) receptor (A(1)R). Mice with a targeted disruption of the Adora 1 gene (A(1)R-/- mice) provide a useful model for better understanding the role of the A(1)R in fertility. Murine spermatozoa express A(1)R in the head, neck, midpiece region, and tail. The number of capacitated spermatozoa incubated in human tubal fluid was significantly reduced in A(1)R-/- compared with A(1)R+/+ and A(1)R+/- spermatozoa. The difference between A(1) R+/+ and A(1)R-/- mouse spermatozoa was mainly in the time necessary to reach the maximum percentage of capacitation. A(1)R+/+ murine sperm obtained the full state of capacitation within 90 minutes whereas A(1)R-/- sperm required 240 minutes. Caffeine, a known antagonist of A(1) and A(2A) adenosine receptors, lowered the number of capacitated sperm and affected the time of capacitation in a dose-dependent manner, mimicking the effects of the lack of A(1) receptors. Although number, motility, and viability of A(1)R-/- murine sperm was not significantly different from A(1)R+/+ mouse spermatozoa, a significant reduction of the number of pups produced by A(1)R-/- male mice suggests that A(1) receptors must be fully operative to accomplish the optimal degree of capacitation and thereby fertilization. PMID- 14760016 TI - The relationship between environmental exposure to phthalates and computer-aided sperm analysis motion parameters. AB - The general population is exposed to phthalates through consumer products, diet, and medical devices. The present study explored whether phthalates, reproductive toxins in laboratory animals, were associated with altered sperm movement characteristics in men. Two-hundred twenty subjects provided a semen sample for computer-aided sperm analysis (CASA) and a urine sample for measurement of phthalate monoesters, monoethyl (MEP), monobenzyl (MBzP), mono-n-butyl (MBP), mono-2-ethylhexyl (MEHP), and monomethyl (MMP). Three CASA parameters, straight line velocity (VSL), curvilinear velocity (VCL), and linearity (LIN), were used as measures of sperm progression, sperm vigor, and swimming pattern, respectively. There were suggestive dose-response relationships (shown as the predicted change in mean sperm motion parameter for the second and third tertiles compared with the first tertile; P value for trend) for MBzP with VSL (-2.36 microm/s, -2.81 microm/s; P =.09) and VCL (-1.67 microm/s, -2.45 microm/s; P =.4). There were suggestive negative associations between MBP and VSL (-3.07 microm/s, -2.87 microm/s; P =.08) and VCL (-3.25 microm/s, -3.46 microm/s; P =.2), and between MEHP with VSL (-1.09 microm/s, -2.73 microm/s; P =.1) and VCL ( 0.29 microm/s, -2.93 microm/s; P =.3). In contrast to the other phthalates, MEP was positively associated with VSL and VCL but negatively associated with LIN. No consistent relationship was found for MMP and any sperm motion parameter. Although we did not find statistically significant associations, trends between CASA parameters, sperm velocity, and forward progression, and increased urinary levels of MBP, MBzP, and MEHP warrant further follow-up. PMID- 14760017 TI - Bridging from research to practice. PMID- 14760018 TI - Reconsidering community based interventions. PMID- 14760019 TI - Driver education renaissance? PMID- 14760020 TI - Longer term effects of New York State's law on drivers' handheld cell phone use. AB - OBJECTIVE: To determine whether substantial short term declines in drivers' use of handheld cell phones, after a state ban, were sustained one year later. DESIGN: Drivers' daytime handheld cell phone use was observed in four New York communities and two Connecticut communities. Observations were conducted one month before the ban, shortly after, and 16 months after. Driver gender, estimated age, and vehicle type were recorded for phone users and a sample of motorists. INTERVENTION: Effective 1 November 2001, New York became the only state in the United States to ban drivers' handheld cell phone use. Connecticut is an adjacent state without such a law. SAMPLE: 50,033 drivers in New York, 28,307 drivers in Connecticut. OUTCOME MEASURES: Drivers' handheld cell phone use rates in New York and Connecticut and rates by driver characteristics. RESULTS: Overall use rates in Connecticut did not change. Overall use in New York declined from 2.3% pre-law to 1.1% shortly after (p<0.05). One year later, use was 2.1%, higher than immediately post-law (p<0.05) and not significantly different from pre-law. Initial declines in use followed by longer term increases were observed for males and females, drivers younger than 60, and car and van drivers; use patterns varied among the four communities. Publicity declined after the law's implementation. No targeted enforcement efforts were evident. Cell phone citations issued during the first 15 months represented 2% of all traffic citations. CONCLUSIONS: Vigorous enforcement campaigns accompanied by publicity appear necessary to achieve longer term compliance with bans on drivers' cell phone use. PMID- 14760021 TI - Risk of hip fracture in protected and unprotected falls in nursing homes in Norway. AB - OBJECTIVES: To compare the probability of hip fracture in protected and unprotected falls in a real world setting in nursing homes. DESIGN: Observational study. SETTING: Seventeen nursing homes (965 beds) in Norway. SUBJECTS: All residents in the nursing homes with at least one fall during the intervention period. INTERVENTION: Hip protectors were introduced as a regular part of the health care service for all the residents for an intervention period of 18 months. Residents who were considered high risk were especially encouraged to be regular users of hip protectors. MAIN OUTCOME MEASURES: Hip fracture in protected and unprotected falls. RESULTS: At the time of the first fall within each faller, 430 were non-users of hip protectors, while 84 were registered as users, but did not wear it, and 191 were users and did wear it. The odds ratio of suffering a hip fracture was 0.31, 95% confidence interval 0.13 to 0.75 for wearers compared with non-wearers in the first fall, adjusted for age, gender, and whether they were registered as users or not. CONCLUSION: The odds of suffering a hip fracture for nursing home high risk residents was reduced to less than a third in protected falls compared with unprotected falls. Or, in other words, the odds of hip fracture showed a 69% reduction in protected falls compared with unprotected falls. PMID- 14760022 TI - Disability and risk of school related injury. AB - OBJECTIVE: Approximately six million children with disabilities attend school in the United States. Cognitive and physical limitations may compromise their ability to handle environmental hazards and hence increase their risk for injury. The objective of this study was to describe the epidemiology of school related injury among children enrolled in 17 special education schools in one large, urban school district. DESIGN: Altogether 6769 schoolchildren with disabilities were followed up from 1994-98. Injury and population data were collected from pupil accident reports and existing school records. Associations were estimated through generalized estimating equations. RESULTS: A total of 697 injuries were reported for a rate of 4.7/100 students per year. Children with multiple disabilities had a 70% increased odds of injury compared with the developmentally disabled (odds ratio (OR) 1.7, 95% confidence interval (CI) 1.3 to 2.3). The physically disabled (OR 1.4, 95% CI 1.0 to 1.9) had a modest increased odds of injury. Cuts, bruises, and abrasions composed almost three fourths of all injuries; almost half of these injuries were to the face. Falls (34%) and insults by other students (31%) were the most common external causes. More than a fourth of injuries were sports related, and 21% occurred on the playground/athletic field. Injury patterns differed across disabilities. CONCLUSIONS: Although limited to one school district, the population studied is the largest cohort thus far of schoolchildren with disabilities. With this large study base, potentially high risk groups were identified and circumstances of injury described. This information is imperative for developing and improving school based injury prevention measures. PMID- 14760023 TI - Are older drivers actually at higher risk of involvement in collisions resulting in deaths or non-fatal injuries among their passengers and other road users? AB - OBJECTIVES: With more older drivers on the road, public concern has been expressed about their impact on traffic safety. This study revisited the question of driver age in relation to the risks of older drivers and others sharing the road with them, including pedestrians, passengers in the same vehicle, and occupants of other vehicles. METHODS: Using United States federal data on fatal and non-fatal crashes, injury rates per driver were calculated for different types of road users. In addition, using data supplied by nine insurers, insurance claims per insured vehicle year were examined by driver age. The reference drivers were aged 30-59. RESULTS: For fatal crashes, older drivers' major impact on road users other than themselves was an increase in death rates among their passengers, who also tended to be elderly and thus more vulnerable to injuries (rate ratio (RR) for drivers aged 75+ 2.52; 95% confidence interval (CI) 2.39 to 2.66). For non-fatal crashes, drivers aged 75+ had a RR of 1.10 (95% CI 0.98 to 1.24) for involvement in collisions resulting in injuries to other passenger vehicles' occupants compared with 30-59 year old drivers. The oldest drivers (aged 85+) had significant increases in insurance claims for injuries to other road users in crashes in which they were deemed at fault (RR 1.8; 95% CI 1.71 to 1.89). CONCLUSIONS: These findings suggest that the oldest drivers, a group with low average annual mileage, do pose some increased risks to occupants of other vehicles, and pose the most serious risks to themselves and their passengers. PMID- 14760024 TI - Motor vehicle driver injury and marital status: a cohort study with prospective and retrospective driver injuries. AB - OBJECTIVE: To investigate the association of marital status with risk of motor vehicle driver injury. DESIGN: A cohort study with prospective and retrospective outcomes. SETTING: New Zealand. PARTICIPANTS: A total of 10,525 adults (a volunteer sample of a multi-industry workforce, n = 8008; and a random sample of urban electoral rolls, n = 2517). EXPOSURE VARIABLE: Self reported marital status, assessed from a questionnaire administered in 1992-93 (baseline). MAIN OUTCOME MEASURE: Motor vehicle driver injury resulting in admission of the driver to hospital and/or the driver's death, during the period 1988-98; hospitalisation and mortality data were obtained by record linkage to national health databases. RESULTS: During 108 741 person-years of follow up, 139 driver injury cases occurred (85 before baseline, 54 after). After adjustment for age, sex, and study cohort, never married participants had twice the risk of driver injury (hazard ratio [HR] 2.06, 95% confidence interval [CI] 1.35 to 3.16) as married participants (HR 1.00). The relative risk for never married participants was slightly higher (HR 2.29), though less precise (95% CI 1.39 to 3.76), after further adjustment for alcohol intake, driving exposure, area of residence, body mass index, and occupational status. CONCLUSIONS: After taking age, sex, and other variables into account, never married people had a substantially higher risk of driver injury than married people. While requiring corroboration, these findings imply that it may be appropriate for driver injury countermeasures to be targeted to never married people. PMID- 14760025 TI - Influence of an injury reduction program on injury and fitness outcomes among soldiers. AB - OBJECTIVE: This study evaluated the influence of a multiple injury control intervention on injury and physical fitness outcomes among soldiers attending United States Army Ordnance School Advanced Individual Training. METHODS: The study design was quasiexperimental involving a historical control group (n = 2559) that was compared to a multiple intervention group (n = 1283). Interventions in the multiple intervention group included modified physical training, injury education, and a unit based injury surveillance system (UBISS). The management responsible for training independently formed an Injury Control Advisory Committee that examined surveillance reports from the UBISS and recommended changes to training. On arrival at school, individual soldiers completed a demographics and lifestyle questionnaire and took an army physical fitness test (APFT: push-ups, sit-ups, and two mile run). Injuries among soldiers were tracked by a clinic based injury surveillance system that was separate from the UBISS. Soldiers completed a final APFT eight weeks after arrival at school. RESULTS: Cox regression (survival analysis) was used to examine differences in time to the first injury while controlling for group differences in demographics, lifestyle characteristics, and physical fitness. The adjusted relative risk of a time loss injury was 1.5 (95% confidence interval 1.2 to 1.8) times higher in the historical control men and 1.8 (95% confidence interval 1.1 to 2.8) times higher in the historical control women compared with the multiple intervention men and women, respectively. After correcting for the lower initial fitness of the multiple intervention group, there were no significant differences between the multiple intervention and historical control groups in terms of improvements in push-ups, sit-ups, or two mile run performance. CONCLUSIONS: This multiple intervention program contributed to a reduction in injuries while improvements in physical fitness were similar to a traditional physical training program previously used at the school. PMID- 14760026 TI - Community based programs to prevent poisoning in children 0-15 years. AB - OBJECTIVE: Community based models for injury prevention have become an accepted part of the overall injury control strategy. This systematic review of the scientific literature examines the evidence for their effectiveness in preventing poisoning in children 0-15 years of age. METHODS: A comprehensive search of the literature was performed using the following study selection criteria: community based intervention study; target population was children under 15 years; outcome measure was poisoning rates; and either a community control or an historical control was used in the study design. Quality assessment and data abstraction were guided by a standardized procedure and performed independently by two authors. Data synthesis was in tabular and text form with meta-analysis not being possible due to the discrepancy in methods and measures between the studies. RESULTS: The review found only four studies, which met all the inclusion criteria. Only two studies used a trial design with a contemporary control and only one study provided convincing evidence of an effective community program for reducing poisoning in children. CONCLUSION: There is a paucity of research studies in the literature from which evidence regarding the effectiveness of community based childhood poisoning prevention programs can be obtained and hence a clear need to increase the effort on developing this evidence base. PMID- 14760027 TI - CDC's National Violent Death Reporting System: background and methodology. AB - OBJECTIVES: This paper describes a new surveillance system called the National Violent Death Reporting System (NVDRS), initiated by the United States Centers for Disease Control and Prevention. NVDRS's mission is the collection of detailed, timely information on all violent deaths. DESIGN: NVDRS is a population based, active surveillance system designed to obtain a complete census of all resident and occurrent violent deaths. Each state collects information on its own deaths from death certificates, medical examiner/coroner files, law enforcement records, and crime laboratories. Deaths occurring in the same incident are linked. Over 270 data elements can be collected on each incident. SETTING: The 13 state health departments of Alaska, Colorado, Georgia, Maryland, Massachusetts, New Jersey, North Carolina, Oklahoma, Oregon, Rhode Island, South Carolina, Virginia, and Wisconsin. SUBJECTS: Cases consist of violent deaths from suicide, homicide, undetermined intent, legal intervention, and unintentional firearm injury. Information is collected on suspects as well as victims. INTERVENTIONS: None. OUTCOME MEASURES: The quality of surveillance will be measured in terms of its acceptability, accuracy, sensitivity, timeliness, utility, and cost. RESULTS: The system has just been started. There are no results as yet. CONCLUSIONS: NVDRS has achieved enough support to begin data collection efforts in selected states. This system will need to overcome the significant barriers to such a large data collection effort. Its success depends on the use of its data to inform and assess violence prevention efforts. If successful, it will open a new chapter in the use of empirical information to guide public policy around violence in the United States. PMID- 14760028 TI - The fast and the fatal: street racing fatal crashes in the United States. AB - The objective of this study was to determine the annual incidence of fatal motor vehicle crashes involving street racing and to describe the characteristics of these crashes compared to other fatal crashes in the United States. The National Highway Traffic Safety Administration Fatality Analysis Reporting System data for 1998-2001 were used for the analyses. There were 149 568 fatal crashes and 315 (0.21%) involved street racing and 399 fatalities occurred in these crashes. In contrast to other fatal crashes, street racing fatal crashes were more likely to occur on urban roadways and were nearly six times more likely to occur at travel speeds> or = 65 mph. Compared with other drivers involved in fatal crashes, street racers were more likely to be teenagers, male, and have previous crashes and driving violations. Street racing involves risky driving behaviors and warrants further attention. PMID- 14760029 TI - Current helmet and protective equipment usage among previously injured ATV and motorcycle riders. AB - Injury recidivism among trauma patients may be related to an individual pattern of high risk behaviors. The extent to which an injury episode modifies this behavior pattern is unknown. A self report, voluntary, anonymous, cross sectional survey was administered to motorcycle and all-terrain vehicle (ATV) riders at a popular recreation site. Data included demographics, injury history, and current usage of helmet and protective gear. Two hundred eighty surveys were completed. History of ATV/motorcycle related minor and major injury were reported by 21% and 9%, respectively. Persons with a history of minor ATV/motorcycle injury only were less likely to use a helmet or protective equipment (78% v 74%, p = 0.58 and 49% v 41%, p = 0.29). Persons with a history of any major ATV/motorcycle injury were also less likely to use a helmet or protective equipment (77% v 56%, p = 0.03 and 48% v 40%, p = 0.53). These findings suggest a pattern of persistent high risk behavior among previously injured persons. PMID- 14760030 TI - Child safety education and the world wide web: an evaluation of the content and quality of online resources. AB - The purpose of this study was to assess the content, quality, and type of internet resources available for safety education. Using 19 search engines with search strings targeting major forms of injury, identified resources were classified by audience group, accessibility, and authorship. Two independent reviewers rated each resource on the basis of its content and a set of quality criteria using a three point scale. Overall, 10 (18.2%) resources were of highest quality, four (7.3%) were intermediate, and 41 (74.5%) were not recommended. Eighteen months after the original search, 67.3% of all resources and 90% of the highest quality resources were still on the internet. This study provides a methodology for evaluating child safety resources on the world wide web and demonstrates that most internet resources for safety education are of dubious quality. A rating system such as the one developed for this study may be used to identify valuable internet materials. PMID- 14760031 TI - Do children's intentions to risk take relate to actual risk taking? AB - OBJECTIVES: Concerns about safety and rigorous ethic standards can make it very difficult to study children's risk taking. The goal of this study was to determine how closely intentions to risk take relate to actual risk taking among boys and girls 6-11 years of age. METHODS: Children initially completed an "intentions to risk take" task. Following administration of several questionnaires they later participated in an actual risk taking task. RESULTS: At all ages, for both boys and girls, intentions to risk take was highly positively correlated with actual risk taking. When discrepancies occurred these were usually of minimal magnitude. CONCLUSIONS: Tasks that tap children's intentions to risk take can serve as proxy indicators of children's actual risk taking. PMID- 14760035 TI - Reporting studies of diagnostic accuracy: the STARD initiative. PMID- 14760036 TI - Towards complete and accurate reporting of studies of diagnostic accuracy: the STARD initiative. AB - OBJECTIVE: Our aim was 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 a study and to evaluate the generalizability of its results. 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 2-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 about 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 a 25-item checklist, by using evidence whenever available. A prototype of a flow diagram provides information about the method of recruitment of patients, the order of test execution and the numbers of patients undergoing the test under evaluation and/or the reference standard. 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 clinicians, researchers, reviewers, journals and the public. PMID- 14760037 TI - Integration of the recommendations of the Canadian Task Force on Preventive Health Care: obstacles perceived by a group of family physicians. AB - BACKGROUND: Surveys conducted in North America and in several European countries show that the preventive activities recommended by some groups of experts are difficult to integrate into medical practice. Interventions to correct this problem have produced mitigated results. OBJECTIVES: Our aim was to gain a better understanding of the obstacles perceived by a group of family physicians concerning the integration of prevention into their routine practices. METHODS: A qualitative design was selected to facilitate the exploration of that topic. Seven focus groups with 35 physicians practising in the Montreal area were conducted. Questions regarding their perception of, and obstacles to, the integration of prevention in their daily work were explored. The text of these interviews was analysed following the content analysis method. Codification of the important themes that were identified was done by two of the researchers. RESULTS: We met with 35 family physicians in two regions in Montreal, Quebec. The lack of motivation on the part of users and the lack of value placed on continuity of care appear to be the main obstacles in the eyes of the physicians, followed by a lack of financial incentives, work overload, and contradictions among the recommendations. In addition, other obstacles were observed by the researchers: limited intervention strategies on the part of physicians to support behaviour modification among patients, non-recognition of the importance of the organization of practice and inability to acknowledge the obstacles that can be ascribed to their own beliefs. CONCLUSION: The family physicians we met identified several barriers to the integration of prevention in their practices. The interventions proposed to date do not address the barriers perceived by the physicians in our study. Continuing medical education activities focus on knowledge, while the difficulties expressed relate more to communication skills coupled with a feeling of powerlessness. The physicians we met with do not seem to consider recall systems and looking at their organization of practice as possible solutions. The physicians seem to 'cave in' under the weight of the responsibilities that have been assigned to them in terms of health promotion. There may be room for proposing a more realistic menu. This study identifies a need for much more specific and concrete training on communication and counselling skills. PMID- 14760038 TI - The value of urine screening in a young adult population. AB - BACKGROUND: GPs in England and Wales are required to perform screening urinalysis on all newly registered patients. The value of this practice, however, is unclear. OBJECTIVE: The purpose of this study was to identify the prevalence of persistent urine abnormalities and to establish the added value of screening for both haematuria and proteinuria in a large cohort of young adults in the UK. METHODS: Urine screening was carried out in a cohort of young adults in a student health centre and a university hospital nephrology unit in a large British city. University students enrolling for health screening in a university health centre over a 2-year period were tested for haematuria and/or proteinuria by dipstick urinalysis. Subjects with persistent urine abnormalities were evaluated for the presence of significant renal tract pathology. RESULTS: Of 3808 students screened, 3570 provided an initial urine sample; 220 were abnormal. Of these, 38 (1% of original cohort) had persistent abnormalities (haematuria, 14; proteinuria, 16; both, eight). Subjects with isolated haematuria or proteinuria did not have significant pathology. In contrast, all the students with both haematuria and proteinuria had identifiable renal disease. CONCLUSIONS: Our findings do not support the value of routine screening for proteinuria or haematuria in young adults. However, the combination of haematuria and proteinuria is a powerful predictor for parenchymal renal disease. Thus, if proteinuria is detected, further testing for haematuria should be performed. PMID- 14760039 TI - Preconception care: practice and beliefs of primary care workers. AB - BACKGROUND: A number of lifestyle modifications and medical interventions can be of benefit to maternal and neonatal health, when applied prior to conception. These include smoking cessation, supplementation with folic acid, cessation or moderation of alcohol intake and improvement of diabetic control. However, preconception care (PCC) is not widely practised in the UK, despite being apparently acceptable to health professionals and to women of childbearing age. OBJECTIVES: The aims of the study were to describe the current practice of PCC in Barnsley and to assess the beliefs and attitudes of primary health care practitioners. This information would help direct appropriate educational and clinical governance intervention to this service in the locality in the light of other evidence about the effectiveness of PCC. METHODS: A questionnaire was devised to explore the beliefs about, and practice in providing, PCC in primary care in the Barnsley Health Authority area and sent to all known GPs, practice nurses (PNs), health visitors (HVs) and midwives (MWs) in practices in the area in July 2000. A total of 163 completed questionnaires were received (one reminder, response rate 60.1%). RESULTS: Few practices had a written policy on PCC. Most respondents were providing it mainly on an opportunistic basis and had done so less than five times in the previous 3 months; GPs and PNs were most commonly involved. They agreed that advice about smoking, drug use, folic acid, genetic counselling, chronic disease, alcohol, and maternity care and screening for rubella, genital infections, hepatitis, human immunodeficiency virus and cervical cytology were important. They felt that advice about diet, exercise, supplements, food safety, occupational hazards and State benefits, and screening for nutritional status were less important. Although respondents felt that PCC was effective, and important to women of childbearing age, it was not a high priority in their workload. They indicated that this care was best provided in general practice and that they had the appropriate skills. Barriers to providing PCC included lack of resources and lack of contact with women planning to conceive. Few had received any training on PCC since qualifying in their discipline. CONCLUSIONS: The practitioners who responded to this survey agreed to a large extent about the importance of the subject, and about the content and effectiveness of PCC. Factors hindering the delivery of this service include resource constraints, lack of training and practice policies and procedures, and difficulty in targeting couples planning conception. Further research is needed into ways to increase the provision and uptake of PCC. PMID- 14760040 TI - You can't prevent everything anyway: a qualitative study of beliefs and attitudes about refusing health screening in general practice. AB - OBJECTIVE: The aim of this study was to explore beliefs and attitudes about refusing health screening in general practice. METHODS: In 1991, in Ebeltoft, Denmark people aged between 30 and 50 years were invited to participate in a 5 year randomized, controlled, population-based project testing the value of health screenings and health discussions in general practice. In 1994, non-participants who declined the offered health screening but expressed willingness to be contacted in the future were asked to participate in a qualitative interview. They were drawn by stratified purposeful sampling which reflected variation in perceived health, body mass index, age and sex. The sample comprised six men and 12 women RESULTS: Some had not participated because they were busy, felt healthy or had recently been examined. The non-participants emphasized the limitations of health screening and did not want possible risk factors to be revealed, or their feeling of good health to be disturbed. They stressed the individual's own responsibility for maintaining good health and believed that a positive attitude promoted health. They would contact their GP if they had symptoms. CONCLUSION: Non-participants have rational views on risk factor testing and on their own responsibility for maintaining health. Non-attendance was due to a conscious choice which included consulting their own GP. PMID- 14760041 TI - Trends in smoking habits: a longitudinal population study. AB - BACKGROUND: In 1998, the UK government published a White Paper in which it set long-term targets for reducing smoking in the population. This longitudinal study aimed to examine whether progress has been made in achieving these in two adult general practice populations over an 8-year period. METHODS: Postal respiratory questionnaires, based on the European Community Respiratory Health Questionnaire, were sent to all patients registered with two practices in North West England on four occasions between 1993 and 2001. Two analyses were carried out. The first (smaller cohort) included only those subjects answering the question concerning current smoking on all four occasions, the second (larger cohort) those answering at least twice. RESULTS: The smaller cohort included 2403 subjects (19.6% of all respondents). Almost one-quarter reported in all four surveys that they smoked, the highest proportion being in those aged 35-44 years. The proportion of smokers decreased from 34.2% (1993) to 30.3% (2001) (P < 0.001 for trend) and the prevalence of heavy smokers fell from 15.9 to 13.3% (P < 0.001 for trend) over the same period. There were, however, no reductions in those aged <45 years in 1993. These changes were confirmed in the larger cohort which included 7274 subjects (59.3% of respondents). CONCLUSIONS: If smoking-related disease is to be reduced, it is important that adults are targeted for smoking prevention and cessation before they reach middle age. Only when such initiatives show success will smoking prevention among their children become a practical proposition. PMID- 14760042 TI - The North Dublin randomized controlled trial of structured diabetes shared care. AB - BACKGROUND: A new diabetes shared care service was introduced in North Dublin. It was designed as a randomized controlled trial with a complex intervention comprising education of participating practitioners, the introduction of a community-based diabetes nurse specialist, local agreement on clinical protocols and structured communication across the primary-secondary care interface. OBJECTIVES: Our aim was to assess the feasibility and effectiveness of a structured diabetes shared care service in a mixed health care system and to analyse the impact on total patient care. METHODS: A Cluster randomized controlled trial lasting 18 months was carried out in 183 patients with type 2 diabetes from 30 general practices in North Dublin. Biophysical outcomes (HbA1c, blood pressure, body mass index), psychosocial measures (smoking status and Diabetes Clinic Treatment Satisfaction and Diabetes Well-being scores) and process outcomes were collected. RESULTS: There were significant improvements in diabetes care delivery and in psychosocial outcomes, but no significant improvements in biomedical outcomes. Process data collection revealed a significant increase in diabetes care-related activity for participating patients with an increase in structured annual reviews and fewer patients defaulting from care. There were also significant improvements in information exchange between primary and secondary care. CONCLUSION: Structured diabetes shared care, in a mixed health care system, can produce significant improvements in diabetes care delivery and in psychosocial outcomes for patients, with improved information exchange across the primary-secondary care interface. PMID- 14760043 TI - What happened to the prescriptions? A single, short, standardized telephone call may increase compliance. AB - BACKGROUND: Patients' compliance with prescribed prescriptions and doctors' advice is a prerequisite for successful treatment. Compliance is estimated to be 70 and BMI >or=33, age >65 and BMI >or=31, age >60 and BMI >or=29, and age >50 and BMI >or=27). Fasting plasma glucose was measured and repeated if abnormal to determine the prevalence of new cases in each group. BMI and age data were validated against measures taken at the clinic. RESULTS: The response rate was 60.6% and the prevalence of new cases of type 2 diabetes in each group was 4.7% [95% confidence interval (CI) 2.8-7.7], 5.7% (95% CI 4.0-8.2), 3.8% (95% CI 2.4 6.0) and 2.6% (95%CI 1.4-4.7), respectively. An additional 5.2-8.4% had IFG. CONCLUSIONS: Targeted screening by searching existing GP records for age and BMI criteria is feasible for use in general practice in the UK. Screening of patients with a BMI of >or=27 and aged >50 by fasting glucose identified a substantial prevalence of undetected type 2 diabetes and IFG. The relative costs and benefits as well as the pragmatic advantages of different systems need further evaluation. PMID- 14760047 TI - A diagnosis of left ventricular hypertrophy on ECG is associated with a high cardiovascular risk: findings from a 40- to 69-year-old cohort in general practice. AB - OBJECTIVE: Left ventricular hypertrophy (LVH) is an independent cardiovascular (CV) risk factor in both sexes. We studied if a diagnosis of LVH on electrocardiogram (ECG) was associated with a 'high CV risk condition' among 40- to 69-year-old individuals cared for by GPs. METHODS: We studied 4250 individuals, 5.4% of whom had LVH. Cross-sectional frequencies, and age- and gender-adjusted statistical differences have been calculated. RESULTS: All the study variables were significantly worse for 'LVH' than 'non-LVH' individuals (except smoking). The 'LVH' had both a mean '5-year CV risk' significantly greater than 'non-LVH' individuals (27.0% versus 8.6%), and a significantly higher prevalence of a '5-year CV risk >15%' (89% versus 15%). CONCLUSIONS: A diagnosis of LVH on ECG among the adult individuals of an opportunistic cohort from general practice was associated with a 6-fold greater prevalence of a 'high CV risk condition'. PMID- 14760048 TI - Is chronic pain a distinct diagnosis in primary care? Evidence arising from the Royal College of General Practitioners' Oral Contraception study. AB - BACKGROUND: and Objectives. There is a case for considering chronic pain as a distinct diagnosis in primary care, independent of its cause or body site, to allow development of common management strategies. This study examined the prevalence and factors associated with chronic pain among women still in the Royal College of General Practitioners' Oral Contraception Study, and compared the factors associated with chronic pain arising at different body sites, to examine the support for this case. METHODS: A survey of 11797 women examined the presence and site(s) of chronic pain, and other health factors. Socio-demographic factors, attendance at the family doctor for the pain and self-rated health were examined for their association with any chronic pain and site-specific chronic pain. RESULTS: Thirty-eight percent of women reported any chronic pain, for which most had attended their family doctor. Manual social class, former oral contraceptive use and area of residence were independently associated with any chronic pain. Chronic pain at different sites was associated with different socio demographic factors, and different proportions of women consulting their family doctor or reporting unfavourable health. These proportions rose with the number of reported sites of chronic pain. CONCLUSIONS: The findings confirm the high prevalence of chronic pain and its importance to primary care. They suggest that chronic pain occurrence at different body sites varies across population subgroups, with differences in its impact on health. These differences call into question the independence of a chronic pain diagnosis. However, other factors common to any chronic pain support the development of common management and prevention strategies in primary care, and the variations highlighted will inform the targeting of these. The number of sites of chronic pain may be more important than the actual site(s) in determining its impact. PMID- 14760049 TI - Do patients and expert doctors agree on the assessment of consultation skills? A comparison of two patient consultation assessment scales with the video component of the MRCGP. AB - OBJECTIVE: The purpose of this study was to determine whether patient ratings of general practice Registrars' consulting skills are associated with 'expert' scoring using the MRCGP video assessment protocol. METHODS: A cross-sectional observational study of general practice Registrars' consultation skills was carried out in 23 practices in South East Scotland using two types of patient assessment compared with expert assessment of video consultation. The main outcome measures were rank correlation of Registrars' overall level of attainment on the Royal College of General Practitioner (RCGP) video assessment with mean score on the Patient Enablement Instrument (PEI) and mean score on the Consultation Satisfaction Questionnaire (CSQ). RESULTS: The rank correlation of Registrars' mean PEI scores with marks on the RCGP video component was 0.01 (P = 0.97, n = 19) and mean CSQ score 0.05 (P = 0.83, n = 19). There were no adverse comments from patients, but Registrars and trainers found the process onerous. CONCLUSION: No meaningful association was identified between Registrars' score on the RCGP video examination and patient assessment via either the PEI or the CSQ. This suggests that, with regard to measuring quality in the consultation, one or more of the assessments are invalid or that they are measuring different attributes. Further research to elucidate the reasons for the lack of correlation is required. PMID- 14760050 TI - Can you compare competence and performance? A reply to McKinstry et al. PMID- 14760051 TI - What happens next? Evaluation of a scheme to support primary care practitioners with a fledgling interest in research. AB - INTRODUCTION: Various initiatives have been implemented in recent years to support primary care practitioners new to research. We describe the evaluation of one particular scheme, the NHS Eastern Region Primary Care R&D Enterprise Award Scheme, and discuss the implications of the results for future policy. METHODS: Representatives from both the funders (NHS R&D) and the award holders were involved in the design of an evaluation questionnaire and in the interpretation of the results. RESULTS: and Discussion. The evaluation demonstrated value in relation to traditional research outcomes and also, notably, in relation to professional contribution. Future policies may need to address: the indicators used in measuring the success of such schemes; the relationship between what individuals choose to do and its context within national policy on research and development; and the sustainability of involvement in research. PMID- 14760052 TI - A case report: ethics of a proposed qualitative study of hospital closure in an Australian rural community. AB - BACKGROUND: The GP and qualitative researcher use similar patient-centred approaches, but their roles are different. Guidelines for conducting GP research in small communities are limited. I planned a qualitative study about hospital closure in a small rural Australian town where I worked. Few studies have researched community reaction to hospital closure and this process of change. METHODS: I used historical analysis to improve external reliability, and purposeful sampling to develop and pre-test a qualitative semi-structured research instrument. Newspaper articles, minutes and tape recordings of public meetings, annual reports from 1991 to 1997, quality assurance data and interviews with two health professionals were analysed in this process. These sources were coded using content and thematic analysis. Ethical issues arose during early stages of planning. Ethical guidelines and bioethics principles were discussed with colleagues and a member of an ethics committee. I validated my findings with three other community members involved in the hospital closure. RESULTS: Themes of a transition, from resistance to change and divisions between key stake holders, to a need to appreciate the benefits of change emerged in coding material from 1991 to 1997. The principle of non-maleficence outweighed the principle of beneficence in this study. Existing health services could be harmed by examining the process of change after spending time and resources to reconcile community differences. Individuals could be harmed as confidentiality in a small community was difficult to maintain, and discussions about sensitive issues could produce adverse public criticism. The autonomy of participants to give informed consent was complicated by the author providing clinical services in the community, raising concerns about patients feeling an obligation to participate. CONCLUSIONS: A justified case for discontinuing this study was made by the researcher on ethical grounds. Use of bioethical principles and community representatives to validate findings was a useful technique to guide decisions in a small rural community. This discussion has application in planning other small community studies. PMID- 14760053 TI - Developing research management and governance capacity in primary care organizations: transferable learning from a qualitative evaluation of UK pilot sites. AB - BACKGROUND: The capacity and capabilities for undertaking primary care research have increased both within and outside of the UK in recent years. The UK Department of Health aims to facilitate this further by establishing a national network of primary care organizations (PCOs) ready to act as hosts for shared research governance systems. However, it is unclear which models offer the most effective option. In addition, there is confusion over new processes and concern that researchers may be deterred from addressing important questions. OBJECTIVES: The research ascertains how PCOs selected as pilot sites have organized research management and governance (RM&G). METHODS: We adopted a case study approach involving interviews with key informants in a purposive sample of eight pilot PCO (RM&G) sites. RESULTS: Motivating factors for PCOs to host RM&G included the possibility of additional resources and more effective use of research to improve service delivery. A range of organizational models were adopted, often reflecting existing strategic alliances. It is envisaged that it will not be effective or cost-effective for many PCOs to make individual arrangements for RM&G, and so models are already developing among groups of PCOs and partner organizations. The extent of partnerships between PCOs varied with concern over critical mass and dilution of expertise in larger groupings. The development and implementation of systems in pilot sites was facilitated by the support of the wider PCO in recognizing research as a valued and integral part of the organization; the effective management of relationships and the establishment of equal partnership arrangements for RM&G, and the effective use of existing R&D infrastructure and expertise. CONCLUSIONS: RM&G partnerships vary according to local circumstances. It is likely that groupings will develop in the future with increasing co terminosity and across wider health organization boundaries, such as Strategic Heath Authorities (in the UK) or primary care research networks. Critical mass of RM&G arrangements is likely to be linked to levels of research activity. There are real concerns over the levels of bureaucracy associated with the implementation of research governance; however, those PCOs that develop as RM&G sites have the opportunity to enrich their organizations and expand clinically relevant R&D. Partnership working within PCOs and with primary care research networks, academic departments or acute trusts, may be the key to success. Those undertaking research within primary care settings outside of the UK can learn important lessons from the UK experience and ensure development of high quality research that informs improvements in patient care. PMID- 14760054 TI - Diagnosis of colorectal cancer in primary care: the evidence base for guidelines. AB - BACKGROUND: Colorectal cancer is common, causing approximately 11% of cancer deaths in the UK. However, a GP would only expect to see one new presentation each year. Referral guidelines outlining clinical scenarios of high risk have been published. These aim to help GPs select patients for rapid investigation. OBJECTIVES: The purpose of this study was to review the presenting features of colorectal cancer in primary care, using the basic structure of the UK Referral Guidelines for Suspected Cancer. METHODS: A structured literature review was carried out. RESULTS: Two symptoms have a high predictive value for cancer: rectal bleeding and change in bowel habit towards increased looseness or increased stool frequency. Other symptoms, such as abdominal pain, are so prevalent in the community that they have little predictive value. There is little published evidence on abdominal or rectal masses and iron deficiency anaemia as presenting features for colorectal cancer. However, these are so likely to have an important cause, investigation is mandated. Two areas in the Referral Guidelines are questioned: the need to defer investigation of change in bowel habit towards increased looseness or increased stool frequency for 6 weeks, and the low risk nature of constipation. CONCLUSION: The Referral Guidelines have a reasonable evidence base. PMID- 14760055 TI - The efficacy of Tai Chi Chuan in older adults: a systematic review. AB - OBJECTIVES: The purpose of this study was to assess the effect of Tai Chi Chuan (TCC) on fall prevention, balance and cardiorespiratory functions in the elderly. METHODS: A systematic review was carried out according to the Cochrane standards. A computerized literature search was carried out. Studies were selected when they had an experimental design; the age of the study population was >50; one of the interventions was a form of TCC; and when falls, balance or cardiorespiratory functions were used as an outcome measure. A total of seven studies were included, with in total 505 participants, of whom all but 27 were healthy seniors, age between 53 and 96 years. RESULTS: In most studies, the intervention of TCC is a modified Yang style, varying from 10 to 24 forms. The intensity of TCC varies from 1 h weekly for 10 weeks to 1 h every morning for 1 year. One study used falls as outcome measure and reported a beneficial effect of 47% in the TCC group. All studies mention a beneficial effect of TCC, but in most studies this conclusion was based on a pre-post analysis. CONCLUSION: There is limited evidence that TCC is effective in reducing falls and blood pressure in the elderly. PMID- 14760056 TI - Doctor-patient communication: emerging challenges. PMID- 14760057 TI - Palliative care in rural areas. PMID- 14760058 TI - The role of thymidylate synthase as a molecular biomarker. PMID- 14760059 TI - Overcoming methodological weakness in dose-intense alkylating agent studies through pharmacology. PMID- 14760060 TI - Vascular targeting agents as cancer therapeutics. AB - Vascular targeting agents (VTAs) for the treatment of cancer are designed to cause a rapid and selective shutdown of the blood vessels of tumors. Unlike antiangiogenic drugs that inhibit the formation of new vessels, VTAs occlude the pre-existing blood vessels of tumors to cause tumor cell death from ischemia and extensive hemorrhagic necrosis. Tumor selectivity is conferred by differences in the pathophysiology of tumor versus normal tissue vessels (e.g., increased proliferation and fragility, and up-regulated proteins). VTAs can kill indirectly the tumor cells that are resistant to conventional antiproliferative cancer therapies, i.e., cells in areas distant from blood vessels where drug penetration is poor, and hypoxia can lead to radiation and drug resistance. VTAs are expected to show the greatest therapeutic benefit as part of combined modality regimens. Preclinical studies have shown VTA-induced enhancement of the effects of conventional chemotherapeutic agents, radiation, hyperthermia, radioimmunotherapy, and antiangiogenic agents. There are broadly two types of VTAs, small molecules and ligand-based, which are grouped together, because they both cause acute vascular shutdown in tumors leading to massive necrosis. The small molecules include the microtubulin destabilizing drugs, combretastatin A-4 disodium phosphate, ZD6126, AVE8062, and Oxi 4503, and the flavonoid, DMXAA. Ligand-based VTAs use antibodies, peptides, or growth factors that bind selectively to tumor versus normal vessels to target tumors with agents that occlude blood vessels. The ligand-based VTAs include fusion proteins (e.g., vascular endothelial growth factor linked to the plant toxin gelonin), immunotoxins (e.g., monoclonal antibodies to endoglin conjugated to ricin A), antibodies linked to cytokines, liposomally encapsulated drugs, and gene therapy approaches. Combretastatin A-4 disodium phosphate, ZD6126, AVE8062, and DMXAA are undergoing clinical evaluation. Phase I monotherapy studies have shown that the agents are tolerated with some demonstration of single agent efficacy. Because efficacy is expected when the agents are used with conventional chemotherapeutic drugs or radiation, the results of Phase II combination studies are eagerly awaited. PMID- 14760061 TI - Searching for reliable epidermal growth factor receptor response predictors: commentary re M. K. Nyati et al., Radiosensitization by pan-ErbB inhibitor CI 1033 in vitro and in vivo. Clin. Cancer Res., 10: 691-700, 2004. PMID- 14760062 TI - Loss of heterozygosity at the thymidylate synthase (TS) locus on chromosome 18 affects tumor response and survival in individuals heterozygous for a 28-bp polymorphism in the TS gene. AB - Thymidylate synthase (TS), the target enzyme of the fluoropyrimidine class of drugs, has a 28-bp repeat polymorphism in the promoter region that has been associated with response of tumors to 5-fluorouracil-based therapy. Patients homozygous for the double repeat (2R/2R) in the TS gene have an overall better outcome from treatment than patients homozygous for the triple repeat (3R/3R). However, due to loss of heterozygosity at the TS locus on chromosome 18 in cancer cells, heterozygous 2R/3R individuals can acquire the 2R/loss or the 3R/loss genotype in their tumors. The purpose of this study was to determine whether the response of colorectal cancer to fluoropyrimidine therapy is associated with the resulting tumor TS genotype when loss of heterozygosity occurs in tumor DNA. A total of 30 colorectal cancer patients treated with the fluoropyrimidine-based combination S-1, all of whom had stage IV disease, were studied. The response rate to S-1 in this group of patients was 13 of 30 (43%). The heterozygous 2R/3R genotype was found in 22 of 30 normal tissues, whereas 10 (45%) of the matched cancer tissues showed only the 2R-sequence band (2R/loss), and 7 cancer tissues (32%) showed only the 3R-sequence band (3R/loss). The response rate of the 2R/loss tumor genotype patients was 80% (8 of 10) compared with 14% (1 of 7) in the 3R/loss genotype group (P = 0.029). Patients with tumor 3R/loss genotypes had significantly lower survival than 2R/loss genotypes. Heterozygous patients with a 2R/loss tumor genotype had the same survival as 2R/2R patients, whereas patients with a 2R/3R tumor genotype had a short survival similar to homozygous 3R/3R genotypes. These results show that: (a) response to 5-fluorouracil-based therapy is determined by tumor genotype; and (b) the 3R repeat is a direct negative determinant of outcome. PMID- 14760063 TI - The androgen axis in recurrent prostate cancer. AB - PURPOSE: Prostate cancer that recurs during androgen deprivation therapy is referred to as androgen-independent. High levels of expression of androgen receptor and androgen receptor-regulated genes in recurrent prostate cancer suggest a role for androgen receptor and its ligands in prostate cancer recurrence. EXPERIMENTAL DESIGN: Recurrent prostate cancer specimens from 22 men whose prostate cancer recurred locally during androgen deprivation therapy and benign prostate specimens from 48 men who had received no prior treatment were studied. Androgen receptor expression was measured using monoclonal antibody and automated digital video image analysis. Tissue androgens were measured using radioimmunoassay. RESULTS: Epithelial nuclei androgen receptor immunostaining in recurrent prostate cancer (mean optical density, 0.284 +/- SD 0.115 and percentage positive nuclei, 83.7 +/- 11.6) was similar to benign prostate (mean optical density, 0.315 +/- 0.044 and percentage positive nuclei, 77.3 +/- 13.0). Tissue levels of testosterone were similar in recurrent prostate cancer (2.78 +/- 2.34 pmol/g tissue) and benign prostate (3.26 +/- 2.66 pmol/g tissue). Tissue levels of dihydrotestosterone, dehydroepiandrosterone, and androstenedione were lower (Wilcoxon, P = 0.0000068, 0.00093, and 0.0089, respectively) in recurrent prostate cancer than in benign prostate, and mean dihydrotestosterone levels, although reduced, remained 1.45 nM. Androgen receptor activation in recurrent prostate cancer was suggested by the androgen-regulated gene product, prostate specific antigen, at 8.80 +/- 10.80 nmol/g tissue. CONCLUSIONS: Testosterone and dihydrotestosterone occur in recurrent prostate cancer tissue at levels sufficient to activate androgen receptor. Novel therapies for recurrent prostate cancer should target androgen receptor directly and prevent the formation of androgens within prostate cancer tissue. PMID- 14760064 TI - Regulating the tumor suppressor gene maspin in breast cancer cells: a potential mechanism for the anticancer properties of tamoxifen. AB - PURPOSE: Mammary epithelial cells and the majority of breast cancer tumors require estrogen for continued growth. Antiestrogen therapy alone, or in combination with other drugs, has long been a common procedure for breast cancer treatment and prophylaxis. Thus, there is a critical need to elucidate the mechanism(s) of action of antiestrogen treatment, especially for patients who are at risk of breast cancer development or who are currently receiving hormone therapy. In this study, we examined the ability of hormones to regulate the expression of a tumor suppressor gene, maspin, which is a serine protease inhibitor (serpin) that plays an important role in mammary gland development and is silenced during breast cancer progression. Specifically, our hypothesis tested the clinical efficacy of tamoxifen to regulate maspin expression. EXPERIMENTAL DESIGN: We used maspin promoter luciferase reporter plasmids that were transfected into normal human mammary epithelial (HMEC1331) and MCF-7 breast cancer cells, followed by determination of the effect of hormones and their antagonists on maspin promoter activity. At the protein level, cytosolic fractions from both cell types before and after hormone treatment were subjected to Western blot analysis to determine maspin level. RESULTS AND CONCLUSIONS: Our studies revealed that the antiestrogen tamoxifen induces maspin promoter activity. Interestingly, antiandrogen flutamide could also induce maspin in both cell lines tested. These observations were further confirmed in patient tissues. These novel findings provide a new mechanism of action for tamoxifen under normal and pathological conditions. More significantly, these findings could have a potential impact on future therapeutic intervention strategies for breast cancer. PMID- 14760065 TI - Cyclophosphamide metabolism in children with non-Hodgkin's lymphoma. AB - PURPOSE: The purpose of our study was to determine whether variation in cyclophosphamide metabolism influences the incidence of recurrence among children receiving chemotherapy for B-cell non-Hodgkin's lymphoma. EXPERIMENTAL DESIGN: The pharmacokinetics and metabolism of cyclophosphamide were studied during a single course of treatment in 36 children receiving a uniform chemotherapy regimen for B-cell non-Hodgkin's lymphoma and were analyzed in terms of disease recurrence and hematological toxicity. RESULTS: At a median follow-up of 43 months (range, 17-98 months), six children had developed recurrent disease, giving an overall disease-free survival of 83%. The median clearance of cyclophosphamide in patients who remain free of B-cell non-Hodgkin's lymphoma was 3.7 liter/h/m(2) (range, 2.3-5.0 liter/h/m(2)), compared with 2.2 (range, 1.5-2.5 liter/h/m(2)) in those with disease recurrence. Likelihood of recurrence was higher in patients with low clearance (<3.5 liter/h/m(2)) of cyclophosphamide (P < 0.01) and positively related to detection of the inactive metabolites carboxyphosphamide and dechloroethylcyclophosphamide in plasma (P = 0.01). There was no correlation between cyclophosphamide metabolism and hematological toxicity. CONCLUSIONS: Inadequate clearance of cyclophosphamide to active metabolites is associated with increased risk of recurrence of B-cell non Hodgkin's lymphoma in children. Modified chemotherapy strategies should be considered in patients who exhibit low rates of clearance of the parent drug and/or extensive production of inactive metabolites. PMID- 14760066 TI - Neurophysiological study of peripheral neuropathy after high-dose Paclitaxel: lack of neuroprotective effect of amifostine. AB - PURPOSE: To determine if there is a beneficial effect of amifostine in preventing or reducing the neuropathy induced by high-dose paclitaxel. METHODS: Breast cancer patients receiving high-dose infusional paclitaxel (725 mg/m(2)/24 h) in combination with doxorubicin (165 mg/m(2)/96 h) and cyclophosphamide (100 mg/kg/2 h; ACT) were studied on two autologous peripheral blood stem cell transplant protocols, one with and one without amifostine (740 mg/m(2) administered over 10 min before and 12 h after initiation of the paclitaxel infusion). Patients were evaluated before ACT and 20-40 days later with neurological examination, a composite peripheral neuropathy score, peroneal and sural nerve conduction studies, and quantitative sensory testing. RESULTS: There was no significant difference in paclitaxel maximum concentration, systemic clearance, or area under the curve determinations. Narcotic requirement as well as recovery of hematopoietic counts were also similar in subjects with or without amifostine. After ACT was administered, there was a decrease in peroneal nerve compound muscle action potential amplitude and sural nerve sensory action potential amplitude, as well as an increase in vibratory and cold detection thresholds. Clinical composite peripheral neuropathy scores were similar despite amifostine treatment; and logarithm to the base 2 ratios post/pre ACT showed no significant effect of amifostine on peroneal nerve compound muscle action potential, sural nerve sensory action potential, vibratory detection thresholds, or cold detection thresholds. All subjects had acroparesthesias and lost their ankle deep-tendon reflexes after administration of ACT. CONCLUSIONS: Single high-dose paclitaxel produces predictable clinical and neurophysiological changes so that patients receiving high-dose therapy are ideal subjects to test the effectiveness of neuroprotective agents. Amifostine was ineffective in preventing or reducing the neurotoxicity of high-dose paclitaxel. PMID- 14760067 TI - A phase I dose-escalation and pharmacokinetic study of brostallicin (PNU 166196A), a novel DNA minor groove binder, in adult patients with advanced solid tumors. AB - PURPOSE: This study was performed to determine the maximum tolerated dose, dose limiting toxicities, and pharmacokinetics of brostallicin, a nonalkylating DNA minor groove binder and a synthetic derivative of distamycin A, given as a weekly i.v. infusion. EXPERIMENTAL DESIGN: Using an accelerated dose escalation design, patients with advanced solid tumor malignancies were treated with brostallicin administered as a 10-min i.v. infusion on days 1, 8, and 15 of a 28-day cycle. The starting dose of brostallicin was 0.3 mg/m(2)/week. To study the pharmacokinetic behavior of brostallicin, serial blood samples were obtained before and after the first and last infusions during cycle 1, and in cycles 2 and 4 in a limited number of patients. RESULTS: Fourteen patients received 32 complete cycles of brostallicin. Dose-limiting toxicity was febrile neutropenia and was observed in 3 of 5 patients treated at 4.8 mg/m(2)/week. The maximum tolerated dose and recommended Phase II dose was 2.4 mg/m(2)/week. The mean +/- SD terminal half-life at the maximum tolerated dose was 4.6 +/- 4.1 h. There was moderate distribution of brostallicin into tissues, and the clearance was approximately 20% of the hepatic blood flow. The area under the concentration time curve(0- infinity ) of brostallicin increased in a dose-linear fashion. No significant relationship was observed between any plasma pharmacokinetic parameter and clinical toxicities. There were no objective responses during the trial, but 5 patients had stable disease after two cycles of treatment. CONCLUSIONS: The dose-limiting toxicity of weekly brostallicin was neutropenia. Systemic exposure increases linearly with dose. The recommended dose for Phase II studies is 2.4 mg/m(2) on days 1, 8, and 15 of a 28-day cycle. PMID- 14760068 TI - Frequency of chromosomal aberrations involving MALT1 in mucosa-associated lymphoid tissue lymphoma in patients with Sjogren's syndrome. AB - PURPOSE: Mucosa-associated lymphoid tissue (MALT) lymphoma develops in the context of longstanding antigenic stimulation such as infection with Helicobacter pylori or autoimmune disease, including Sjogren's syndrome (SS). Recently, two chromosomal aberrations involving the MALT1 gene, i.e., t(11;18)(q21;q21) and t(14;18)(q32;q21) have been reported as genetic events specific for MALT lymphoma. In view of the association between SS and the development of MALT lymphoma, we have analyzed the frequency of t(11;18)(q21;q21) and t(14;18)(q32;q21) in patients with MALT lymphomas arising in the background of SS. EXPERIMENTAL DESIGN: A retrospective analysis of patients diagnosed with MALT lymphoma and SS was performed. The t(11;18)(q21;q21) was analyzed using reverse transcriptase-PCR, whereas t(14;18)(q32;q21) was assessed by two-color interphase fluorescence in situ hybridization. RESULTS: Twenty-six patients (20 female and 6 male) with MALT lymphoma and SS could be identified. The lymphoma was located in the parotid (n = 14), orbit (n = 2), and submandibular gland (n = 1), whereas 9 patients had gastric MALT lymphoma. Seven of 26 patients (27%) harbored t(11;18)(q21;q21). Interestingly, only 1 of 17 patients (6%) with extragastrointestinal lymphoma was positive, as opposed to 6 of 9 patients (67%) with gastric MALT lymphoma. Four of 26 patients were positive for t(14;18)(q32;q21): 3 of 17 extragastrointestinal (18%) and 1 of 9 gastric lymphomas (11%). CONCLUSIONS: The overall frequency of MALT1 rearrangement appears to be low in patients with extragastrointestinal MALT lymphoma associated with SS. By contrast, MALT1 rearrangement was demonstrated in 7 of 9 patients (78%) with gastric MALT lymphoma and SS. This finding may explain at least in part why gastric MALT lymphomas in patients with SS are refractory to H. pylori eradication therapy. PMID- 14760069 TI - Increased risk for hereditary nonpolyposis colorectal cancer-associated synchronous and metachronous malignancies in patients with microsatellite instability-positive endometrial carcinoma lacking MLH1 promoter methylation. AB - PURPOSE: The aim of this study was to evaluate number and types of synchronous and metachronous malignancies in patients with endometrial carcinoma with and without microsatellite instability (MSI). EXPERIMENTAL DESIGN: From a series of 413 endometrial cancer patients, we identified 94 patients with MSI-positive (MSI+) cancers and grouped them by tumor MLH1 promoter methylation status. These 94 patients were matched by year of surgery to 94 patients with MSI-negative (MSI ) endometrial cancers from the same series. Medical records were reviewed for clinicopathologic information including rates and types of synchronous and metachronous malignancies. Hereditary nonpolyposis colorectal cancer (HNPCC) associated second and third cancers were analyzed for MSI and MSH2, MSH6, and MLH1 expression for comparison with the corresponding endometrial cancers. RESULTS: The MSI+ and MSI- cohorts were similar with regard to age, race, grade, and histology. Twenty-eight MSI+ endometrial cancers (29.8%) were MLH1 unmethylated. Rates of synchronous and metachronous cancers were also similar in the MSI+ and MSI- groups at 20 and 23%, respectively. However, patients with MSI+ MLH1 unmethylated endometrial cancers had an excess of HNPCC-associated second and third cancers compared with those with MSI+ MLH1 methylated and MSI- endometrial cancers (18% versus 4.5%, P = 0.034, and 2.1%, P = 0.002). Six of seven second tumors from 5 patients with MSI+ MLH1 unmethylated endometrial cancers showed concordant MSI and mismatch repair protein expression status. CONCLUSIONS: Our observation that patients with MSI-positive MLH1 unmethylated endometrial carcinoma are at increased risk for HNPCC-associated synchronous and metachronous malignancies suggests inherited cancer susceptibility. These patients and their families may warrant more intense cancer surveillance. PMID- 14760070 TI - Prognostic significance of transforming growth factor beta receptor II in estrogen receptor-negative breast cancer patients. AB - PURPOSE: The role of transforming growth factor beta (TGF-beta) in breast cancer is ambiguous; it can display both tumor suppressing and enhancing effects. Activation of the TGF-beta signal transduction system is subject to hormonal regulation. This study was conducted to further analyze the role of TGF-beta receptors in breast cancer and to evaluate their significance as prognostic markers. EXPERIMENTAL DESIGN: Expression of TGF-beta receptor I (TbetaRI) and TGFbeta receptor II (TbetaRII) was retrospectively analyzed by immunohistochemistry in 246 breast cancer patients. RESULTS: Expression of TbetaRI was strongly correlated with tumor size (P < 0.001) and nodal status (P = 0.012) but only weakly with overall survival (P = 0.056). In contrast, TbetaRII was prognostic for overall survival in univariate analysis (P = 0.0370). In estrogen receptor (ER) -negative patients TbetaRII expression was correlated with highly reduced overall survival (P = 0.0083). In multivariate analysis TbetaRII proved to be an independent and highly significant prognostic marker with a hazard ratio of 6.8. Simultaneous loss of both ER and TbetaRII was associated with longer overall survival times comparable with those of ER-positive patients. CONCLUSIONS: The results of this exploratory study show that TbetaRII is an independent, highly significant prognostic indicator for overall survival in ER negative patients. In addition our results are supportive of a mechanism of breast cancer progression in which a selective loss of the tumor inhibitory action of TGFbeta takes place, whereas tumor- promoting aspects remain intact. PMID- 14760071 TI - MYC is amplified in BRCA1-associated breast cancers. AB - PURPOSE: Germ-line mutations in the BRCA1 tumor suppressor gene predispose to early onset breast cancers with a distinct phenotype characterized by high tumor grade, aneuploidy, high proliferation rate, and estrogen receptor-negativity. The molecular mechanisms and cooperative oncogenes contributing to multistep tumor progression in cells lacking BRCA1 are not well defined. To examine whether C-MYC (MYC), a transforming oncogene associated with genetic instability, contributes to multistep tumor progression in BRCA1-associated breast cancer, we have analyzed tumors from women with hereditary BRCA1-mutated and sporadic breast cancers. EXPERIMENTAL DESIGN: We performed fluorescence in situ hybridization using a MYC:CEP8 assay on formalin-fixed paraffin-embedded tumor tissues from 40 women with known deleterious germ-line BRCA1 mutations and 62 sporadic cases, including 20 cases with hypermethylation of the BRCA1 gene promoter. RESULTS: We observed a MYC:CEP8 amplification ratio >/=2 in 21 of 40 (53%) BRCA1-mutated tumors compared with 14 of 62 (23%) sporadic tumors (P = 0.003). Of the 14 sporadic cases with MYC amplification, 8 (57%) were BRCA1-methylated. In total, MYC amplification was found in a significantly higher proportion of tumors with BRCA1 dysfunction (29 of 60, 48% versus 6 of 42, 14%; P = 0.0003). In a multivariable regression model controlling for age, tumor size, and estrogen receptor status, BRCA1-mutated tumors demonstrated significantly greater mean MYC:CEP8 ratio than sporadic tumors (P = 0.02). CONCLUSIONS: Our data indicate that MYC oncogene amplification contributes to tumor progression in BRCA1 associated breast cancers. Thus, we conclude that the aggressive histopathological features of BRCA1-associated tumors are in part due to dysregulated MYC activity. PMID- 14760072 TI - CD26 expression correlates with a reduced sensitivity to 2'-deoxycoformycin induced growth inhibition and apoptosis in T-cell leukemia/lymphomas. AB - PURPOSE AND EXPERIMENTAL DESIGN: dCF (2'-deoxycoformycin) is a potent inhibitor of ADA (adenosine deaminase), an enzyme regulating intra- and extracellular concentrations of purine metabolites. ADA exists as cytosolic and extracellular forms, the latter colocalized on the cell surface with CD26. Once the surface expression of CD26 and ADA in a panel of cell lines and primary samples of T-cell leukemia/lymphoma was defined, we correlated this expression with the antiproliferative and apoptotic effect of dCF. RESULTS: Surface expression of CD26 inversely correlated with the capability of dCF to inhibit cell growth and induce apoptosis both in T-cell lines and primary samples of T-cell malignancies. This conclusion was sustained by a decreased sensitivity to dCF-mediated proapoptotic and/or antiproliferative in vitro effects of: (a) leukemia/lymphoma T-cell lines expressing surface CD26/ADA complex; (b) primary CD26(+) T cell malignancies; and (c) normal T cells (CD26(+)) as compared with tumor T cells (CD26(-)) in unpurified samples from three cases of T-cell receptor gammadelta(+) T-cell malignancies characterized by a mixture of normal and neoplastic cells. This latter point was confirmed in vivo, in a patient affected by CD26(-) T-cell receptor gammadelta(+) hepatosplenic gammadelta(+) T-cell lymphomas treated on a compassionate basis with dCF. The inverse correlation between CD26 expression and sensitivity to dCF was also demonstrated in a lymphoblastic lymphoma case in which CD26 was expressed on circulating blasts at relapse but not at diagnosis, as well as in two H9 T-cell clones expressing or not expressing CD26 mRNA and protein. CONCLUSIONS: This study corroborates the notion of CD26 as a marker of poor prognosis for T-cell malignancies and delineates a role for CD26 as a predictor of poor response to dCF. PMID- 14760073 TI - T-cell activation marker expression on tumor-infiltrating lymphocytes as prognostic factor in cutaneous malignant melanoma. AB - The central role of T cells in antitumor immunity is well established. However, tumor progression, often seen in the presence of substantial lymphocytic infiltration, suggests that these T cells are not capable of mounting an effective immune response to control tumor growth. Evidence has accumulated that T lymphocytes infiltrating human neoplasms are functionally defective, incompletely activated, or anergic. Therefore, when characterizing the immune competent cells within lymphoid infiltrates of tumors, it is important to assess their activation state. We investigated the expression of two T-cell activation markers, interleukin 2 receptor alpha (CD25) and OX40 (CD134), by immunohistochemistry in primary cutaneous melanoma samples of 76 patients and analyzed it in relation to tumor stage and tumor progression (>5 years follow up), as well as to patients' survival. We found that the degree of infiltration by CD25(+) and intratumoral OX40(+) lymphocytes showed a tendency to decrease in thicker melanomas. The frequency of samples with high numbers of peritumoral CD25(+) and OX40(+) cells was significantly lower (P = 0.0009 and P = 0.0087, respectively) in melanomas developing distant visceral metastases, compared with nonmetastatic or lymph node metastatic tumors. For both activation markers studied, high peritumoral densities were associated with longer survival by univariate analysis (P = 0.0028 and P = 0.0255 for CD25 and OX40, respectively), whereas peritumoral OX40(+) lymphocyte infiltration had an impact on survival also in multivariate analysis (P = 0.035). The results suggest that the presence of lymphocytes expressing the T-cell activation markers CD25 or OX40 shows correlation with tumor progression as well as with patients' survival in cutaneous malignant melanoma. PMID- 14760074 TI - Immunomagnetic enrichment, genomic characterization, and prognostic impact of circulating melanoma cells. AB - PURPOSE: The finding of melanoma cells in the peripheral blood, thus far mainly inferred from the PCR-based demonstration of tyrosinase mRNA, has been associated with metastatic melanoma. Neither the malignant nature nor the prognostic significance of circulating cells could be established. To address this question, we analyzed immunomagnetically isolated circulating melanoma cells for chromosomal aberrations and performed a clinical follow-up study of the enrolled patients. EXPERIMENTAL DESIGN: In a prospective study, blood samples were taken from 164 melanoma patients and 50 donors without malignant disease. Circulating melanoma cells were enriched by immunomagnetic cell sorting using a murine monoclonal antibody against the melanoma-associated chondroitin sulfate proteoglycan. To prove the malignant origin of the positive cells and to define their chromosomal aberrations, we analyzed the genomes of 15 individually isolated cells from seven patients by single-cell comparative genomic hybridization (SCOMP). RESULTS: Absolute and relative frequencies of circulating melanoma cells were associated with stage and with the presence or absence of detectable tumor. The detection of two or more cells correlated significantly with a reduced survival of patients with metastatic melanoma. All of the cells that were analyzed by SCOMP displayed multiple chromosomal changes and carried aberrations typical for melanoma. CONCLUSIONS: Immunomagnetic enrichment enables isolation and genomic characterization of circulating melanoma cells. The prognostic impact on survival of metastatic patients apparently reflects the aggressiveness of an ongoing tumor spread. Direct genomic analysis of the enriched and isolated cells will help to clarify the molecular-genetic basis of the establishment of generalized melanoma. PMID- 14760075 TI - Elevated cyclooxygenase-2 expression is associated with altered expression of p53 and SMAD4, amplification of HER-2/neu, and poor outcome in serous ovarian carcinoma. AB - PURPOSE AND EXPERIMENTAL DESIGN: Cyclooxygenase-2 (COX-2) is frequently expressed in human adenocarcinomas and inhibition of COX-2 suppresses tumor formation in various animal models of carcinogenesis. We analyzed expression of COX-2 protein in human serous ovarian carcinomas by immunohistochemistry (n = 442) and by Western blotting (n = 12) and COX-2 mRNA by reverse transcriptase PCR (n = 12). COX-2 immunoreactivity was correlated to clinicopathological variables and to expression of p53 and SMAD4 as detected by immunohistochemistry and to amplification of HER-2/neu as detected by in situ hybridization. RESULTS: COX-2 mRNA expression was detected in 75% (9 of 12) and COX-2 protein in 42% (5 of 12) of the serous ovarian adenocarcinoma specimens as detected by reverse transcriptase-PCR and Western blot analysis, respectively. Moderate to strong (elevated) immunoreactivity for COX-2 was detected in 70% (310 of 442) of the tumors. Elevated COX-2 expression associated with reduced disease-specific survival (P = 0.0011), high histological grade (P < 0.0001), residual tumor size > 1 cm (P = 0.0111), and age > 57 years (P = 0.0099). Tumors with altered immunostaining pattern for p53 or SMAD4 expressed more frequently elevated levels of COX-2 when compared with the tumors with normal staining pattern of these tumor suppressor genes (P < 0.0001 and P = 0.0004, respectively). In addition, elevated COX-2 expression associated with amplification of HER-2/neu oncogene (P = 0.0479). CONCLUSIONS: Our results suggest that elevated expression of COX-2 associates with reduced survival in serous ovarian carcinomas and that expression of COX-2 may be induced in these tumors by loss of tumor suppressor genes such as p53 and SMAD4 and by amplification of HER-2/neuoncogene. PMID- 14760076 TI - SRC kinase and mitogen-activated protein kinases in the progression from normal to malignant endometrium. AB - PURPOSE: The purpose of this research was to determine whether a correlation exists between the levels of activated mitogen-activated protein kinase (MAPK) and Src kinases and the progression from normal to malignant endometrium. EXPERIMENTAL DESIGN: We measured total and phosphorylated levels for extracellular signal-regulated kinase 1/2, p38, stress-activated protein kinase/c Jun NH(2)-terminal kinase, and Src kinases from 33 frozen endometrial adenocarcinomas and 38 benign endometrial specimens by quantitation of signals from Western blots using antibodies against these kinases. RESULTS: Elevated phospho-extracellular signal-regulated kinase 1/2 (150 +/- 40 versus 46 +/- 7; P = 0.03), phospho-Src (28 +/- 5 versus 4 +/- 1), and phospho-p38 (131 +/- 16 versus 27 +/- 7; P < 0.001) was detected in benign versus malignant endometrium when the Western blot signal of activated kinase was normalized to total kinase levels and beta actin. A modest increase in active c-Jun NH(2)-terminal kinase was detected in carcinoma versus benign specimens (51 +/- 13 versus 43 +/- 10; P = 0.8). Expression of total kinases (normalized to beta-actin) was higher in carcinoma versus benign specimens, respectively (extracellular signal-regulated kinase 1/2, 9 +/- 2 versus 0.7 +/- 0.1; Src, 7 +/- 2 versus 0.4 +/- 0.1; stress activated protein kinase c-Jun NH(2)-terminal kinase, 2 +/- 0.4 versus 0.2 +/- 0.02; P < 0.001; and p38, 1 +/- 0.2 versus 0.4 +/- 0.1; P < 0.01). Immunohistochemistry for active and total Src kinases and MAPKs detected positive staining in epithelial and stroma cells. CONCLUSIONS: These data demonstrated that, in contrast with breast cancer, the progression from normal to malignant endometrium is not associated with activation of MAPK and Src kinases. Elevation of these active kinases in benign endometrium may contribute to endometrial resistance to the antiestrogen action of tamoxifen. PMID- 14760077 TI - Microsatellite instability predicts poor short-term survival in patients with advanced breast cancer after high-dose chemotherapy and autologous stem-cell transplantation. AB - PURPOSE: The purpose is to define molecular prognostic factors in patients with advanced breast cancer treated with high-dose chemotherapy (HDCT) and autologous stem cell transplantation (ASCT). EXPERIMENTAL DESIGN: Thirty-nine patients with breast cancer and extensive lymph node (level III) and/or systemic metastases from a prospective single-center study of sequential HDCT/ASCT were studied. Microsatellite analysis was performed after laser microdissection using 15 markers selected for sensitive detection of microsatellite instability (MSI) in breast cancer. Exons 5-9 of the P53 gene were directly sequenced. Expression of P53, HER-2/neu, and the mismatch repair proteins hMSH2 and hMLH1 was evaluated by immunohistochemistry. RESULTS: MSI of at least three markers was detected in 13 of 39 patients (33%) and was predominantly found at tetranucleotide markers. All MSI-positive tumors showed normal expression of hMSH2 and hMLH1. Complete sequence analysis of exons 5-9 of the P53 gene was successful in 34 cases; 18% (n = 6) revealed a mutation. Overexpression of HER-2/neu and P53 was observed in 7 (22%) and 12 (46%) of 26 evaluated cases, respectively. The presence of MSI strongly correlated with shorter overall survival (OS; P = 0.0004) and progression-free survival (PFS; P = 0.02). None of the other investigated clinical or molecular factors correlated with OS in univariate analyses, with the exception of menopausal status and previous adjuvant chemotherapy. Testing various multivariate Cox regression models, MSI remained a highly significant, independent, and adverse risk factor for OS. CONCLUSIONS: MSI is frequent in advanced breast cancer and could be an indicator of chemotherapy resistance and poor prognosis in breast cancer patients treated with HDCT/ASCT. PMID- 14760078 TI - DNA methylation in serum and tumors of cervical cancer patients. AB - PURPOSE: Promoter hypermethylation has been recognized to play an important role in carcinogenesis. Numerous studies have demonstrated tumor-specific alterations, such as aberrant promoter hypermethylation, in DNA recovered from plasma or serum of patients with various malignancies. The aim of this study was to investigate the methylation status of various genes in cervical cancer patients and their association with clinicopathological characteristics and outcome of the disease. EXPERIMENTAL DESIGN: The methylation status of CALCA, hTERT, MYOD1, PGR (progesterone receptor), and TIMP3 was investigated in serum samples from 93 cervical cancer patients and 19 corresponding tissue samples using the MethyLight technique. RESULTS: Aberrant promoter hypermethylation was detected in any of these genes in 87% (81 of 93) of the serum samples studied. Methylation of MYOD1 was detected more frequently in advanced stage. All of the genes found to be methylated in serum samples were also methylated in the corresponding tissue sample, except in one patient. Patients with unmethylated MYOD1 serum DNA had significantly better disease-free (P = 0.04) and overall survival (P = 0.02) in comparison with patients with methylated MYOD1. CONCLUSIONS: To the best of our knowledge, this is, thus far, the largest study investigating aberrant promoter hypermethylation in serum samples from cancer patients and the first study investigating methylation patterns in sera of cervical cancer patients. Our results suggest that serological detection of MYOD1 promoter hypermethylation may be of potential use as a prognostic marker for discriminating cervical cancer patients at high risk for lymph node metastasis or relapse. Additional studies, including a panel of additional genes, are necessary to elucidate the role of aberrant methylation in serum as a tool for surveillance of cervical cancer. PMID- 14760079 TI - Clinical significance of cellular distribution of moesin in patients with oral squamous cell carcinoma. AB - PURPOSE: Moesin is a linking protein of the submembraneous cytoskeleton and plays a key role in the control of cell morphology, adhesion, and motility. The aim of the present study was to elucidate the clinical significance of expression patterns of moesin in patients with oral squamous cell carcinoma (OSCC). EXPERIMENTAL DESIGN: Immunohistochemistry for moesin monoclonal antibody was performed on 103 paraffin-embedded specimens from patients with primary OSCC, including 30 patients with locoregional lymph node metastasis, and in the sections from nude mice transplanted with two cell lines derived from a single human tongue cancer (SQUU-A and SQUU-B). RESULTS: Expression patterns of moesin in OSCCs were divided into three groups: membranous pattern; mixed pattern; and cytoplasmic pattern. These expression patterns correlated with tumor size, lymph node metastasis, mode of invasion, differentiation, and lymphocytic infiltration. In about two-thirds of the patients with metastatic lymph node, homogeneous cytoplasmic expression was detected in the metastatic lymph nodes. In addition, SQUU-B with high metastatic potential showed more reduced levels of membrane bound moesin than SQUU-A with low metastatic potential. A multivariate analysis demonstrated that expression patterns of moesin can be an independent prognostic factor. CONCLUSIONS: Our results suggest that moesin expression contributed to discriminating between patients with the potentiality for locoregional lymph node metastasis and those with a better prognosis and might improve the definition of suitable therapy for each. PMID- 14760080 TI - Aberrant expression of neuropilin-1 and -2 in human pancreatic cancer cells. AB - PURPOSE: Neuropilin (Np)-1 and -2 are coreceptors for vascular endothelial growth factor (VEGF). This study was designed to assess their role in pancreatic ductal adenocarcinoma (PDAC). EXPERIMENTAL DESIGN: We assessed Np-1 and Np-2 expression by real-time quantitative PCR in relation to the expression of VEGF ligands and receptors in pancreatic cancer cell lines and tissues. RESULTS: ASPC-1, CAPAN-1, and PANC-1 pancreatic cancer cells and tumor-derived, laser-captured pancreatic cancer cells exhibited higher Np-1 and Np-2 mRNA levels than VEGF receptor-1, -2, or -3 mRNA levels. Transfection of Np-1 and Np-2 cDNAs in COS-7 cells, and treatment with tunicamycin revealed that both proteins were glycosylated. Both proteins were expressed in pancreatic cancer cell lines, in the PDAC samples, and in acinar cells adjacent to the cancer cells. The normal pancreas was devoid of Np-1 immunoreactivity, whereas Np-2 immunoreactivity was present in the endocrine islets and in some acinar cells, but not in ductal cells. CONCLUSIONS: The aberrant localization of Np-1 and Np-2 in the cancer cells in PDAC suggests that in addition to exerting proangiogenic effects, these coreceptors may contribute to novel autocrine-paracrine interactions in this malignancy. PMID- 14760081 TI - MIB-1 (KI-67) proliferation index and cyclin-dependent kinase inhibitor p27(Kip1) protein expression in nephroblastoma. AB - PURPOSE: A number of studies have indicated that the tumor proliferation marker MIB-1 and cell cycle inhibitor p27(Kip1) expression are of prognostic importance in a variety of cancers. The present study was performed to evaluate the prognostic value of these molecules in Wilms' tumors. EXPERIMENTAL DESIGN: MIB-1 and p27(Kip1) expressions were investigated by the means of immunohistochemical analysis of 62 Wilms' tumor. Patients were preoperatively treated by chemotherapeutic agents and had a mean follow-up of 5.7 years. RESULTS: MIB-1 and p27(Kip1) were expressed in normal kidney tissues and in the three main components of Wilms' tumor, i.e., the blastemal, epithelial, and stromal cells. In Wilms' tumors, the percentage of MIB-1-positive cells in the blastema ranged between 0 and 42% (mean, 9.4%) and in the epithelial component between 0 and 53% (mean, 19.9%), with a significant difference (P < 0.01). The percentage of blastemal p27(Kip1)-positive cells ranged between 3 and 85% (mean, 55.1%) and for the epithelial component between 1 and 87% (mean, 59%). There was a significant inverse relationship between blastemal MIB-1 and p27(Kip1) expression in Wilms' tumor. Univariate analysis showed that blastemal MIB-1 and p27(Kip1) expression were indicative for clinical progression and tumor-specific survival. In a multivariate analysis, blastemal MIB-1 and p27(Kip1) protein expression proved to be an independent prognostic for clinical progression besides stage. CONCLUSIONS: It was concluded that both MIB-1-based proliferative activity and p27(Kip1) protein expression in the blastema have prognostic impact in Wilms' tumor. PMID- 14760082 TI - Enhanced protein profiling arrays with ELISA-based amplification for high throughput molecular changes of tumor patients' plasma. AB - PURPOSE: The purpose of this study is to develop a high-throughput approach to detect protein expression from hundreds and thousands of samples and to apply this technology to profile circulating angiogenic factor protein levels in patients with gynecological tumors. EXPERIMENTAL DESIGN: Analytes containing a mixture of protein are immobilized onto antibody-coated surface of support in array format. The presence of protein in analytes is detected with biotin-labeled antibody coupled with an enhanced chemiluminescence or fluorescence detection system. The exact amount of protein can be quantitatively measured. The expression levels of five angiogenic factors (angiogenin, interleukin 8, vascular endothelial growth factor, platelet-derived growth factor, and epidermal growth factor) from 157 samples were quantitatively measured using this novel protein array technology and were statistically analyzed. The expression patterns of angiogenic factors were analyzed using two-way hierarchical cluster analysis approach. RESULTS: A novel protein array technology, which can simultaneously and quantitatively measure few protein levels from hundreds and thousands of samples was developed. Only minute amounts of sample are required for the assay. This approach also features high sensitivity and specificity. Using this novel protein array approach, we analyzed the plasma expression levels of five angiogenic factors in 137 patients diagnosed with a tumor and 20 controls. Statistical analysis reveals different expression levels of angiogenic factors between patients and controls. Cluster analysis suggests a possible classification of normal subjects from patients. CONCLUSIONS: Enhanced protein profiling arrays provide a high-throughput and sensitive system to detect one or few protein from hundreds and thousands of samples. Such an approach should have broad application in biomedical discovery. PMID- 14760083 TI - Soluble CD40 ligand plasma levels in lung cancer. AB - PURPOSE: Tumor-induced platelet activation may cause the release of various cytokines, including CD40 ligand (CD40L). Activation of the CD40/CD40L pathway in human tumors may result in thrombin generation, which is known to be involved in angiogenesis. Thus, we investigated whether soluble (s)CD40L levels are increased in patients with lung cancer as a result of platelet and/or coagulation activation. EXPERIMENTAL DESIGN: Citrated plasma samples were obtained from 120 patients with different stages and histotypes of lung cancer and 60 age- and sex matched control subjects. sCD40L, sP-selectin (marker of platelet activation), prothrombin fragment 1 + 2, and thrombin-antithrombin III complex levels (both markers of coagulative activation) were measured in all samples. RESULTS: Patients with lung cancer had median sCD40L levels higher than in control subjects (0.46 versus 0.13 ng/ml; P < 0.0001), although correlation with the stage of disease was not evident. Nonetheless, sCD40L levels were significantly higher in squamous cancer compared with adenocarcinoma (0.75 versus 0.27 ng/ml; P < 0.05). Moreover, median sCD40L levels were higher in stage IV compared with nonmetastatic squamous lung cancer (1.02 versus 0.61 ng/ml; P < 0.05). sCD40L levels significantly correlated with sP-selectin (P < 0.001), prothrombin fragment 1 + 2 (P < 0.001), or thrombin-antithrombin III complex (P < 0.05) in squamous lung cancer, but only sP-selectin (P = 0.011) was independently related to sCD40L. CONCLUSIONS: These findings indicate that elevated sCD40L levels can be preferentially found in patients with advanced squamous cancer and provide evidence that increased levels of this cytokine are associated to the occurrence of in vivo platelet activation. PMID- 14760084 TI - Membranous expression of secreted frizzled-related protein 4 predicts for good prognosis in localized prostate cancer and inhibits PC3 cellular proliferation in vitro. AB - PURPOSE: Activation of the Wnt-signaling pathway is implicated in aberrant cellular proliferation in a variety of cancers. Secreted frizzled-related protein 4 (sFRP4) is a secreted protein with putative inhibitory activity of the Wnt signaling cascade through binding and sequestering Wnt ligands. Because sFRP4 mRNA is overexpressed in prostate cancers (PCs), the aim of this study was to define the pattern of sFRP4 protein expression in normal and malignant human prostate tissue and to determine whether changes in expression were associated with disease progression and prognosis, as well as to define the phenotype of sFRP4-overexpression in an in vitro model of PC. EXPERIMENTAL DESIGN: Polyclonal antibodies were raised against a COOH-terminal peptide of sFRP4, characterized and used to assess sFRP4 protein expression in benign prostate tissue and 229 patients with clinically localized PC (median follow-up 77 months, range 1-156). In vitro studies of the function of sFRP4 overexpression were performed using PC3 cells transfected with sFRP4. RESULTS: Benign and malignant prostate tissue demonstrated cytoplasmic sFRP4 immunoreactivity, but there was a decrease in the expression of membranous sFRP4 in PCs compared with the hyperplastic lesions (P < 0.0001). Kaplan-Meier analysis revealed that patients whose PC expressed membranous sFRP4 in >20% of cells had improved relapse-free survival compared with those with /==" BORDER="0">2-fold change in only 6 (0.027%) and 51 (0.23%) of 33,000 genes surveyed, respectively. No changes were seen in expression of genes involved in cell cycle, apoptosis, DNA repair, or recombination. CONCLUSIONS: These results demonstrate that telomestatin exerts its antiproliferative and proapoptotic effects in myeloma cells via inhibition of telomerase and subsequent reduction in telomere length. We conclude that telomerase is an important potential therapeutic target for multiple myeloma therapy, and G-quadruplex interacting agents with specificity for binding to telomeric sequences can be important agents for additional evaluation. PMID- 14760101 TI - Interleukin-12 enhances the sensitivity of human osteosarcoma cells to 4 hydroperoxycyclophosphamide by a mechanism involving the Fas/Fas-ligand pathway. AB - Cyclophosphamide (CY) and its derivative ifosfamide are alkylating agents used to treat osteosarcoma (OS). The purpose of these studies was to determine whether alkylating agents affect the expression of Fas ligand (FasL) and whether interleukin 12 enhances the sensitivity of human OS cells to alkylating agents. 4 Hydroperoxycyclophosphamide (4-HC), the preactivated CY compound, and 4 hydroperoxydidechlorocloclophosphamide (4-HDC), its nonalkylating analogue, human OS LM6 cells, and a clone of cells derived by transfection with the interleukin 12 gene (LM6-#6) were used for these studies. Incubation of LM6 and LM6-#6 with 10 micro M 4-HC increased the expression of FasL mRNA (2.5- and 3.0-fold, respectively). By contrast, 4-HDC, Adriamycin (ADR), cisplatin (CDP), and methotrexate (MTX) had no effect on FasL mRNA expression. Increased FasL expression after treatment with 4-HC was also demonstrated by immunohistochemistry and flow cytometry. Drug-induced FasL was functional and mediated cell death. We examined the effect of FasL up-regulation by 4-HC on LM6 and LM6-#6 cells. Flow cytometry showed that LM6-#6 cells expressed 2.2-fold more Fas than LM6 cells. Cytotoxicity of 4-HC, 4-HDC, ADR, CDP, and MTX on LM6, LM6 neo, and LM6-#6 were quantified. Colony-forming assay revealed an IC(50) of 2.10 micro M for 4-HC in LM6-neo cells compared with 0.41 micro M in LM6-#6 cells. The IC(50) for 4-HDC, ADR, CDP, and MTX were not significantly different between the two cell lines. We concluded that the increased expression of Fas enhanced LM6-#6 sensitivity to 4-HC. These data indicate that Fas/FasL may be involved in the cytotoxic pathway of CY. Combining biological agents with chemotherapeutic agents that have complementary Fas/FasL pathway actions may offer new therapeutic alternatives. PMID- 14760102 TI - Antitumor activity of ZD6474, a vascular endothelial growth factor receptor tyrosine kinase inhibitor, in human cancer cells with acquired resistance to antiepidermal growth factor receptor therapy. AB - PURPOSE: The epidermal growth factor receptor (EGFR) autocrine signaling pathway is involved in cancer development and progression. EGFR inhibitors such as C225 (cetuximab), a chimeric human-mouse anti-EGFR monoclonal antibody, and ZD1839 (gefitinib), a small molecule EGFR-selective tyrosine kinase inhibitor, are in advanced clinical development. The potential emergence of cancer cell resistance in EGFR-expressing cancers treated with EGFR inhibitors could determine lack of activity of these drugs in some cancer patients. Vascular endothelial growth factor (VEGF) is secreted by cancer cells and plays a key role in the regulation of tumor-induced endothelial cell proliferation and permeability. ZD6474 is a small molecule VEGF flk-1/KDR (VEGFR-2) tyrosine kinase inhibitor that also demonstrates inhibitory activity against EGFR tyrosine kinase. EXPERIMENTAL DESIGN: The antitumor activity of ZD1839, C225, and ZD6474 was tested in athymic mice bearing human GEO colon cancer xenografts. GEO cell lines resistant to EGFR inhibitors were established from GEO xenografts growing in mice treated chronically with ZD1839 or C225. Expression of EGFR was evaluated by flow cytometry. Expression of various proteins involved in intracellular cell signaling was assessed by Western blotting. Tumor growth data were evaluated for statistical significance using the Student's t test. All Ps were two-sided. RESULTS: Although chronic administration of optimal doses of C225 or ZD1839 efficiently blocked GEO tumor growth in the majority of mice, tumors slowly started to grow within 80-90 days, despite continuous treatment. In contrast, continuous treatment of mice bearing established GEO xenografts with ZD6474 resulted in efficient tumor growth inhibition for the entire duration of dosing (up to 150 days). ZD6474 activity was also determined in mice pretreated with ZD1839 or C225. When GEO growth was apparent after 4 weeks of treatment with EGFR inhibitors, mice were either re-treated with EGFR inhibitors or treated with ZD6474. GEO tumor growth was blocked only in mice treated with ZD6474, whereas tumor progression was observed in mice re-treated with C225 or ZD1839. GEO tumors growing during treatment with C225 or with ZD1839 were established as cell lines (GEO-C225-RES and GEO-ZD1839-RES, respectively). Cell membrane-associated EGFR expression was only slightly reduced in these cell lines compared with parental GEO cells. Western blotting revealed no major change in the expression of the EGFR ligand transforming growth factor alpha of bcl-2, bcl-xL, p53, p27, MDM-2, akt, activated phospho-akt, or mitogen-activated protein kinase. However, both GEO-C225-RES and GEO-ZD1839-RES cells exhibited a 5-10-fold increase in activated phospho-mitogen-activated protein kinase and in the expression of cyclooxygenase 2 and of VEGF compared with GEO cells. GEO-C225-RES and GEO-ZD1839-RES growth as xenografts in nude mice was not significantly affected by treatment with either C225 or ZD1839 but was efficiently inhibited by ZD6474. CONCLUSIONS: Long-term treatment of GEO xenografts with selective EGFR inhibitors results in the development of EGFR inhibitor-resistant cancer cells. Growth of EGFR inhibitor resistant tumors can be inhibited by ZD6474. These data indicate that inhibition of VEGF signaling has potential as an anticancer strategy, even in tumors that are resistant to EGF inhibitors. PMID- 14760103 TI - ZD1839 modulates paclitaxel response in renal cancer by blocking paclitaxel induced activation of the epidermal growth factor receptor-extracellular signal regulated kinase pathway. AB - PURPOSE: We evaluated the antitumor activity of ZD1839, a selective epidermal growth factor receptor (EGFR)-tyrosine kinase inhibitor, in combination with paclitaxel in human renal cell carcinomas (RCCs). EXPERIMENTAL DESIGN: Eight human RCC lines and the surgical specimens obtained from 10 RCC patients were used. The protein expression was detected by Western blotting, immunohistochemistry and/or flow cytometry. Apoptosis was evaluated by flow cytometry and fragmented DNA ELISA. SKRC-49 tumor xenografts in athymic nude mice were treated with ZD1839 and/or paclitaxel, and tumor volume was determined RESULTS: EGFR protein was expressed and phosphorylated in eight RCC lines and EGFR expression was markedly increased in RCC specimens compared with adjacent normal renal tissues. Treatment of SKRC-49 with 1 micro M ZD1839 resulted in a marked decrease in the phosphorylation of EGFR but not of HER-2. Treatment of SKRC-49 with ZD1839 in combination with 5 nM paclitaxel resulted in a significant increase in apoptotic cell number compared with paclitaxel alone, whereas ZD1839 alone failed to induce apoptosis. Although administration of ZD1839 or paclitaxel resulted in a transient growth inhibition in SKRC-49 xenografts, significant tumor regrowth delay was observed when paclitaxel was combined with ZD1839. Paclitaxel phosphorylated extracellular signal-regulated kinase through EGFR activation predominantly in cancer cells. ZD1839 promoted paclitaxel-induced Bcl 2 down-regulation resulting in promoting apoptosis by blocking paclitaxel-induced activation of the EGFR-extracellular signal-regulated kinase antiapoptotic pathway independent of Akt activity in SKRC-49. CONCLUSIONS: Our findings support the idea that the significant clinical benefit is obtained from ZD1839 in combination with paclitaxel for the treatment of RCC. PMID- 14760104 TI - Correspondence re: C. Ford, et al. Mouse mammary tumor virus-like gene sequences in breast tumors of Australian and Vietnamese women. Clin. Cancer Res., 9: 1118 1120, 2003. PMID- 14760106 TI - Evaluation of renal function in elderly cancer patients. PMID- 14760108 TI - Human papillomavirus vaccine as a new way of preventing cervical cancer: a dream or the future? AB - Cervical cancer is the major cause of death in women of reproductive age in parts of the developing world. Thanks to the effectiveness of national screening programs, the incidence and mortality rates for cervical cancer have declined dramatically in developed countries. According to many researchers, human papillomavirus (HPV) infection has an important role in the development of cervical neoplasm. The effects of HPV infection on the oncogenesis of cervical carcinoma can be explained to a large degree by the regulation and function of the two viral oncogenes, E6 and E7. About 25 of >80 types infect the genital tract. HPV types are stratified into low, intermediate- and high-risk categories. Today, vaccines are available against many serious human pathogens. It is accepted worldwide that cervical carcinoma is a consequence of infection with HPV viruses. Therefore it is reasonable to assume that vaccine that prevents infection will reduce the incidence of cervical cancer. Virus-like particles are empty viral capsids, and are the leading candidate vaccines for the treatment or prevention of cervical cancer in humans. The HPV type 16 (HPV16) L1 virus-like particle vaccines have been shown to be generally well tolerated and they generate high levels of antibodies against HPV16. Since approximately 50% of cervical cancers are associated with HPV16 infection, the administration of this type of vaccine to young women could reduce the incidence of HPV16 infection, which is related to cervical dysplasia and cervical neoplasm. Vaccination against HPV infection could reduce the risk of infection and, most importantly, decrease the incidence of cervical cancer. A vaccine for cervical cancer is not a dream in the far future, it is happening today. PMID- 14760107 TI - Chemoprevention of lung cancer--from biology to clinical reality. AB - Lung cancer is the commonest cause of cancer death in developed countries and throughout the world. Cigarette smoking is the main risk factor for lung cancer and ex-smokers today comprise approximately 50% of all new lung cancer cases. Chemoprevention builds on the concepts of field of cancerization and multistep carcinogenesis and can be defined as the use of natural or chemical compounds to prevent, inhibit or reverse the process of carcinogenesis. So far, chemoprevention studies in lung cancer have failed to reduce lung cancer mortality. New developments in biotechnology have made it possible to define more accurately high-risk populations, make earlier diagnosis possible, and allow more specific targeted therapies to be developed. Both the development and validation of biomarkers, for the selection of high-risk study populations and for response evaluation in chemoprevention studies, are important for the faster turnover of studies evaluating new agents. This article reviews the current status and describes the perspectives for new approaches in the chemoprevention of lung cancer. PMID- 14760109 TI - Biweekly paclitaxel plus gemcitabine in advanced breast cancer: phase II trial and predictive value of HER2 extracellular domain. AB - BACKGROUND: We wanted to assess the toxicity and efficacy of paclitaxel plus gemcitabine in advanced breast cancer and to confirm whether circulating HER2 extracellular domain (ECD) correlates with treatment response. PATIENTS AND METHODS: Forty-three patients received paclitaxel 150 mg/m2 followed by gemcitabine 2500 mg/m2, both on day 1 of 14-day cycles, with a maximum of eight cycles. Serum levels of HER2 ECD were assessed by ELISA. RESULTS: All patients were evaluable for toxicity and 42 for efficacy. Overall toxicity was low. Grade 3 neutropenia occurred in 12% of patients and grade 4 in 17%, and other grade 3 toxicities in <5%. One patient had an allergic infusion reaction. Overall response rate was 71% [95% confidence interval (CI) 62% to 81%], with 11 patients achieving a complete response (26%). With a median follow-up of 26 months, the median time to progression was 16.6 months. Response rate correlated significantly with HER2 ECD, with 42% of HER2 ECD-positive patients responding versus 83% of HER2 ECD-negative patients (P = 0.02). Furthermore, response duration was shorter in patients with positive HER2 ECD levels (7.9 versus 14.4 months; P = 0.04). CONCLUSIONS: Paclitaxel plus gemcitabine given as an every 2 weeks schedule is a well tolerated and active regimen in advanced breast carcinoma. This is an attractive combination to use when anthracyclines are not indicated, such as in HER2 positive cases that receive trastuzumab. In addition, elevated levels of HER2 ECD adversely affect the efficacy of treatment. PMID- 14760110 TI - Adjuvant chemotherapy in elderly patients with breast cancer: a survey of the Breast International Group (BIG). AB - BACKGROUND: To collect oncologists' experience and opinion on adjuvant chemotherapy in elderly breast cancer patients. MATERIALS AND METHODS: A questionnaire was circulated among the members of the Breast International Group. RESULTS: A total of 277 oncologists from 28 countries participated in the survey. Seventy years is the age cut-off commonly used to define a patient as elderly. Biological age and the biological characteristics of the tumor are the most frequently used criteria to propose adjuvant chemotherapy to an elderly patient. Combination therapy with cyclophosphamide, methotrexate and fluorouracil on days 1 and 8 is the most frequently prescribed regimen. Great interest exists in oral chemotherapy. CONCLUSION: There is interest among those who responded to the survey to validate a comprehensive geriatric assessment for use as a predictive instrument of toxicity and/or activity of anticancer therapy and to evaluate the role of a treatment option that is potentially less toxic and possibly as effective as polychemotherapy. PMID- 14760111 TI - The effect of exemestane on serum lipid profile in postmenopausal women with metastatic breast cancer: a companion study to EORTC Trial 10951, 'Randomized phase II study in first line hormonal treatment for metastatic breast cancer with exemestane or tamoxifen in postmenopausal patients'. AB - BACKGROUND: The impact of aromatase inhibitors (AIs) on non-cancer-related outcomes, which are known to be affected by oestrogens, has become increasingly important in postmenopausal women with hormone-dependent breast cancer. So far, data related to the effect of AIs on lipid profile in postmenopausal women is scarce. This study, as a companion substudy of an EORTC phase II trial (10951), evaluated the impact of exemestane, a steroidal aromatase inactivator, on the lipid profile of postmenopausal metastatic breast cancer (MBC) patients. PATIENTS AND METHODS: The EORTC trial 10951 randomised 122 postmenopausal breast cancer patients to exemestane (E) 25 mg (n = 62) or tamoxifen (T) 20 mg (n = 60) once daily as a first-line treatment in the metastatic setting. Exemestane showed promising results in all the primary efficacy end points of the trial (response rate, clinical benefit rate and response duration), and it was well tolerated with low incidence of serious toxicity. As a secondary end point of this phase II trial, serum triglycerides (TRG), high-density lipoprotein cholesterol (HDL), total cholesterol (TC), lipoprotein a (Lip a), and apolipoproteins (Apo) B and A1 were measured at baseline and while on therapy (at 8, 24 and 48 weeks) to assess the impact of exemestane and tamoxifen on serum lipid profiles. Of the 122 randomised patients, those who had baseline and at least one other lipid assessment are included in the present analysis. The patients who received concomitant drugs that could affect lipid profile are included only if these drugs were administered throughout the study treatment. Increase or decrease in lipid parameters within 20% of baseline were considered as non-significant and thus unchanged. RESULTS: Seventy-two patients (36 in both arms) were included in the statistical analysis. The majority of patients had abnormal TC and normal TRG, HDL, Apo A1, Apo B and Lip a levels at baseline. Neither exemestane nor tamoxifen had adverse effects on TC, HDL, Apo A1, Apo B or Lip a levels at 8, 24 and 48 weeks of treatment. Exemestane and tamoxifen had opposite effects on TRG levels: exemestane lowered while tamoxifen increased TRG levels over time. There were too few patients with normal baseline TC and abnormal TRG, HDL, Apo A1, Apo B and Lip a levels to allow for assessment of E's impact on these subsets. The atherogenic risk determined by Apo A1:Apo B and TC:HDL ratios remained unchanged throughout the treatment period in both the E and T arms. CONCLUSIONS: Overall, exemestane has no detrimental effect on cholesterol levels and the atherogenic indices, which are well-known risk factors for coronary artery disease. In addition, it has a beneficial effect on TRG levels. These data, coupled with E's excellent efficacy and tolerability, support further exploration of its potential in the metastatic, adjuvant and chemopreventive setting. PMID- 14760112 TI - Value of routine follow-up procedures for patients with stage I/II cervical cancer treated with combined surgery-radiation therapy. AB - BACKGROUND: The aim of this study was to determine the value of routine follow-up for the detection of recurrence in patients treated for cervical cancer. PATIENTS AND METHODS: From 1986 to 1998, 583 women with stage I and II cervical carcinoma were treated with combined surgery-radiation therapy. After treatment, follow-up was based on clinical examination, a systematic Pap smear and radiography (chest X-ray and abdomino-pelvic ultrasonography). RESULTS: Forty-five patients had recurrence observed with a delay > or = 6 months following the end of treatment. Thirty-eight patients had symptoms and seven were asymptomatic at the time of their recurrence. Among asymptomatic patients only two recurrences were diagnosed following routine examinations. Survival is similar in asymptomatic and symptomatic recurrent patients. CONCLUSIONS: In conclusion, follow-up of patients treated for cervical cancer based on routine Pap smears and systematic radiography does not permit earlier detection of recurrence and does not increase survival. PMID- 14760113 TI - Front-line treatment of inoperable or metastatic pancreatic cancer with gemcitabine and capecitabine: an intergroup, multicenter, phase II study. AB - PURPOSE: To evaluate the efficacy and toxicity of gemcitabine (GEM) combined with capecitabine (CAP) in untreated patients with inoperable or metastatic pancreatic cancer. PATIENTS AND METHODS: Fifty-three patients with pancreatic cancer (85% stage IV) were enrolled. Patients were treated with GEM 1000 mg/m2 on days 1 and 8 and CAP 1300 mg/m2 per day PO (per os), divided into two equal doses on days 1 14, in 21-day cycles. RESULTS: In an-intention-to-treat analysis, 10 (18.9%) objective partial responses were achieved (95% confidence interval 8.33% to 29.4%). Twenty-two (42%) patients had stable disease and 15 (28%) had progressive disease. The median response time was 3 months (range 1.5-7.0) and the median time to tumor progression was 6.5 months (range 3.5-15.5). Median overall survival time was 8 months (range 1.0-15.5) and 1-year survival was 34.8%. Pain improvement during treatment was observed in 23 of 43 (53%) patients, and eight of 18 (44%) patients who had been receiving opioids discontinued their use. Weight gain was observed in 12 of 33 (36%) patients. Grade 3 anemia occurred in five (9%) patients and grade 3-4 thrombocytopenia occurred in three (6%). Grade 3 4 neutropenia occurred in 13 (25%) and five (9%) patients, respectively, and two (4%) developed febrile neutropenia. Non-hematological toxicity was mild. CONCLUSION: In patients with pancreatic cancer, the combination of GEM with CAP is an active and well tolerated regimen that merits further evaluation in prospective randomized studies. PMID- 14760114 TI - Phase II study of cisplatin preceding gemcitabine in patients with advanced oesophageal cancer. AB - BACKGROUND: For oesophageal cancer there is no effective standard therapy. We studied the feasibility and efficacy of the cisplatin-gemcitabine combination chemotherapy in patients with unresectable oesophageal cancer. PATIENTS AND METHODS: Thirty-six chemonaive patients with unresectable or metastatic oesophageal adenocarcinoma (24) or squamous-cell-carcinoma (12) were treated with cisplatin (50 mg/m2, days 1 and 8), followed by gemcitabine (800 mg/m2, days 2, 9 and 16), every 28 days. Feasibility and efficacy were studied. RESULTS: Toxicity was substantial but manageable. A median number of four therapy cycles was given. The most frequent grade > or = 3 toxicities were leukopenia (75%) and neutropenia (83%). Three patients developed neutropenic fever. Grade 3/4 thrombocytopenia occurred in 24 out of 36 patients (67%), but did not result in serious bleeding disorders. Myelotoxicity was cumulative and required omission of gemcitabine on day 16 in 63% of cycles. Anaemia required treatment with erythropoietin, red blood cells or both in 81% of patients. Non-haematological toxicity consisted mainly of grade 1/2 nausea/vomiting or fatigue. Fourteen out of 34 evaluable patients had a major objective response (41%; two complete and 12 partial responses). The median actuarial survival was 9.8 months. CONCLUSION: This cisplatin-gemcitabine regimen was feasible, with myelosupression being the main toxicity, and had significant activity in patients with advanced oesophageal cancer. PMID- 14760115 TI - Clinical significance of the overexpression of the candidate oncogene CYP24 in esophageal cancer. AB - BACKGROUND: By using array comparative genomic hybridization (CGH), the increased copy number of CYP24 (which encodes vitamin D 24-hydroxylase) at 20q13.2 was previously reported, leading to the identification of CYP24 as a candidate oncogene in breast cancer. CYP24 leads to abrogate growth control mediated by vitamin D. MATERIALS AND METHODS: We examined CYP24 expression as well as VDR (vitamin D receptor) gene expression in 42 esophageal cancer cases using semi quantitative RT-PCR assay. We induced CYP24 in seven esophageal cancer cell lines using 25-hydroxyvitamin D3 [25(OH)D3] and compared cell growth rate, measured using the 3-(4, 5-dimethylthiazol-2-y)-2, 5-diphenyltetrazolium bromide (MTT) assay system. RESULTS: The overall survival rate was significantly higher in 25 cases of lower CYP24 expression than 17 cases of higher CYP24 expression (P <0.05); on the other hand, 23 cases of low VDR expression had a poorer prognosis than 19 cases of high VDR expression. Moreover, we disclosed that the inverse correlation between CYP24 and VDR expression is significant in esophageal cancer cases (P <0.05). Furthermore, the cell growth evaluated by MTT assay was greatly increased in CYP24-induced and VDR-diminished cells than non-responding cells by 25(OH)D3 activity (P <0.01). CONCLUSIONS: Overexpression of the candidate oncogene CYP24 is inversely correlated to vitamin D receptor expression, and may play an important role in determination of the malignant potential of esophageal cancer. PMID- 14760116 TI - Clinicopathological significance of microsatellite instability and mutated RIZ in colorectal cancer. AB - BACKGROUND: Several studies have shown that microsatellite instability (MSI) is related to favourable survival in colorectal cancer patients but there are controversial results. Tumour suppressor gene RIZ is a susceptible mutational target of MSI. However, its clinicopathological significance has not been investigated. We investigated the prognostic significance of MSI in Swedish colorectal cancer patients and the clinicopathological significance of RIZ mutations. PATIENTS AND METHODS: We analysed 438 colorectal adenocarcinomas for MSI by microsatellite analysis. Among them, 29 MSI and 28 microsatellite stable (MSS) tumours were examined for RIZ mutations by DNA sequencing. RESULTS: MSI (13% of 438 cases) was not associated with survival (rate ratio = 0.97, 95% confidence interval = 0.57-1.64, P = 0.90), although it was related to proximal tumour (P <0.001), poor differentiation and mucinous carcinomas (P <0.001), multiple tumours (P = 0.01) and negative/weak expression of hMLH1 (P = 0.03). RIZ mutations were detected in 31% of 29 MSI tumours but in none of the 28 MSS tumours. The mutations were related to female (P = 0.01), proximal tumour (P = 0.01), stage B (P = 0.01) and poor differentiation (P = 0.047). CONCLUSIONS: MSI was not a prognostic factor in the Swedish patients included in this study. Clinicopathological variables associated with RIZ mutations might be a consequence of the MSI characteristics. PMID- 14760117 TI - Phase II study of topotecan and paclitaxel for patients with previously untreated extensive stage small-cell lung cancer. AB - BACKGROUND: This phase II study was conducted to evaluate the safety and efficacy of the combination of paclitaxel and topotecan for patients with extensive stage small-cell lung cancer (ED-SCLC). PATIENTS AND METHODS: Previously untreated ED SCLC patients with Eastern Coperative Oncology Group performance status <2 were eligible. Treatment consisted of topotecan 1 mg/m2 (first three patients received 1.25 mg/m2), on days 1-5, and paclitaxel 135 mg/m2 over 24 h on day 5, every 4 weeks. Prophylactic granulocyte colony-stimulating factor was administered to all patients. RESULTS: Thirty-two patients received a median of four cycles of chemotherapy. Grade 4 anemia, neutropenia and thrombocytopenia occurred in 13, 31 and 18 patients, respectively. Thirty episodes of febrile neutropenia occurred in 22 patients. Grade 3 fatigue, esophagitis, stomatitis and hypotension occurred in one patient each. Of 26 patients eligible for response evaluation, there were six complete and 12 partial responses (overall response rate 69%). The median survival was 54 weeks. The 1-, 2- and 3-year survival rates were 50%, 10% and 3%, respectively. CONCLUSIONS: The combination of topotecan and paclitaxel is active as initial therapy in SCLC, but the efficacy is similar to 'standard therapy'. This combination was associated with a high incidence of myelosuppression and febrile neutropenia, at the doses evaluated. PMID- 14760118 TI - The clinical significance of Cyclin B1 and Wee1 expression in non-small-cell lung cancer. AB - BACKGROUND: Cyclin B1 has an important role in the G2-M phase transition of the cell cycle. Wee1 delays mitosis by suppressing the activity of the Cyclin B1/cdc2 complex. The objective of the present study was to elucidate the clinicopathological and prognostic significance of Cyclin B1 and Wee1 expression in non-small-cell lung cancer (NSCLC). PATIENTS AND METHODS: An immunohistochemical assessment of Cyclin B1 and Wee1 expression was performed in 79 patients with NSCLC. RESULTS: The expression of Cyclin B1 was correlated with differentiation (P = 0.0423) and vascular invasion (P = 0.001). Patients with overexpression of Cyclin B1 had higher mean values for both the Ki-67 proliferative index (Ki-Index) (P <0.0001) and proliferating cell nuclear antigen labeling index (PCNA-LI) (P <0.0001), and a poorer prognosis (P = 0.0068). Patients lacking expression of Wee1 had a higher recurrence rate (P = 0.0084) and a poorer prognosis (P = 0.0457), and tended to have higher Ki-Index and PCNA-LI values. Multivariate analysis suggested that both Cyclin B1 (P = 0.0244) and Wee1 (P = 0.0444) expression were significant prognostic factors. CONCLUSIONS: These findings suggest that Cyclin B1 expression could be a significant prognostic parameter in NSCLC. The loss of Wee1 expression may have a potential role in promoting tumor progression and may be a significant prognostic indicator in NSCLC. PMID- 14760119 TI - Modified RECIST criteria for assessment of response in malignant pleural mesothelioma. AB - BACKGROUND: The growth pattern of malignant pleural mesothelioma makes the use of RECIST (response evaluation criteria in solid tumours) response criteria difficult. We have developed and validated Modified RECIST criteria adapted to the growth pattern of malignant pleural mesothelioma. PATIENTS AND METHODS: We evaluated 73 patients from two clinical trials of cisplatin/gemcitabine chemotherapy in malignant pleural mesothelioma. Tumour thickness perpendicular to the chest wall or mediastinum was measured in two positions at three separate levels on thoracic CT scans. The sum of the six measurements defined a pleural unidimensional measure. Bidimensionally measureable lesions were measured unidimensionally as for RECIST. All measurements were added to obtain the total tumour measurement. A reduction of at least 30% on two occasions 4 weeks apart defined a partial response; an increase of 20% over the nadir measurement, progressive disease. The validity of the modified criteria was gauged by evaluating survival and pulmonary function. RESULTS: Response according to these criteria predicted for superior survival (15.1 versus 8.9 months; P = 0.03) and forced vital capacity (FVC) increase during treatment (P <0.0001). A significant correlation between change in linear tumour measurement and FVC was seen (R = 0.63; P = 0.0001). CONCLUSION: These Modified RECIST criteria for tumour response correlate with survival and lung function and can be used to measure outcome in pleural mesothelioma. PMID- 14760120 TI - Factors influencing treatment recommendations in early-stage Hodgkin's disease: a survey of physicians. AB - BACKGROUND: The aim of this study was to explore variation in practice patterns and identify factors associated with physicians' treatment decisions for early stage Hodgkin's disease. METHODS: We conducted a one-time mail survey of oncologists randomly selected from directories of national oncology societies (n = 207) and Hodgkin's disease experts (n = 147). The survey included questions on (i) physician factors, (ii) preferred treatment choices for six case scenarios of early-stage Hodgkin's disease that varied by patient factors, and (iii) thresholds for changing treatment recommendations. RESULTS: The response rate was 50%. For non-bulky Hodgkin's disease, 69% of respondents chose combined modality therapy (CMT). On multivariate analysis, physician factors that independently predicted for choice of CMT included a high Hodgkin's disease case load (P = 0.02) and a high percentage of patients enrolled in clinical trials (P = 0.05). Radiation oncologists had a lower threshold for adding radiation therapy (P = 0.02). More experience with second malignancy cases and longer time in practice were associated with a higher threshold for adding radiation therapy (P = 0.04 and P = 0.008, respectively). In stratified analyses, treatment decisions of non experts were significantly influenced by physician factors, but not by patient factors. Conversely, choices of Hodgkin's disease experts were insensitive to all physician factors, but experts were significantly more likely to select chemotherapy alone in young women and CMT in older patients. CONCLUSIONS: Our results indicate that physician factors including practice type and experience may in part explain variation in practice pattern for Hodgkin's disease therapy. Hodgkin's disease experts are more likely to tailor therapy according to individual patient factors. PMID- 14760121 TI - Comparison of MOPP versus ABVD as salvage therapy in patients who relapse after radiation therapy alone for Hodgkin's disease. AB - BACKGROUND: The aim of this study was to determine salvage outcome in patients with Hodgkin's disease who relapse after radiation therapy, and to compare the efficacy of mechlorethamine, Oncovin, procarbazine and prednisone (MOPP) versus Adriamycin, bleomycin, vinblastine and dacarbazine (ABVD) as salvage treatment. PATIENTS AND METHODS: One hundred patients with Hodgkin's disease (97 with stage I-II disease at presentation) who relapsed after radiation therapy alone were salvaged with either MOPP or ABVD. Freedom from second relapse (FFSR) and overall survival (OS) were determined, and prognostic factors for salvage outcome were evaluated. RESULTS: The median follow-up time since salvage therapy was 12 years. The 10-year FFSR and OS rates were 70% and 89%, respectively. Forty-one patients were salvaged with MOPP and 59 received ABVD. The type of salvage chemotherapy did not significantly influence FFSR or OS. Age >50 years at initial diagnosis was the only significant predictor for an inferior FFSR and OS on both univariate and multivariate analyses. CONCLUSIONS: The two salvage regimens of MOPP and ABVD had similar efficacy in this group of patients with predominantly early-stage disease at initial radiation therapy. The inferior salvage outcome in patients aged >50 years is a contributing factor to the overall poor prognosis of patients presenting with Hodgkin's disease at an older age. PMID- 14760122 TI - Treatment of advanced Hodgkin's disease with COPP/ABV/IMEP versus COPP/ABVD and consolidating radiotherapy: final results of the German Hodgkin's Lymphoma Study Group HD6 trial. AB - BACKGROUND: The purpose of this study was to compare the efficacy of the hybrid chemotherapeutic regimen COPP/ABV/IMEP (cyclophosphamide-vincristine-procarbazine prednisone-doxorubicin-bleomycin-vinblastine-ifosfamide-methotrexate-etoposide) (CAI) with that of the standard regimen COPP/ABVD (COPP/ABV, dacarbacine) (CA) in the treatment of advanced-stage Hodgkin's disease (HD). PATIENTS AND METHODS: Between January 1988 and January 1993, 588 eligible patients with HD in stages IIIB and IV were randomly assigned to a treatment or control group. The treatment group received four cycles of CAI over a complete cycle duration of 43 days. The control group received four cycles of CA over 57 days. Both groups then received consolidating radiotherapy. RESULTS: Five hundred and eighty-four patients were suitable for arm comparison. Patients in each group were similar in age, sex, histological subtype and clinical risk factors. Complete remission rates, overall survival and freedom from treatment failure at 7 years were similar for the two groups: 77% versus 78%, 73% versus 73% and 54% versus 56% for CAI and CA, respectively. Differences in acute chemotherapy-related toxicity were significant, however. Prognostic factor analysis confirmed the relevance of the International Prognostic Index and revealed that stage IVB, low hemoglobin, low lymphocyte count, high age and male gender were associated with a poor prognosis CONCLUSION: The rapidly alternating hybrid CAI did not give superior results when compared with the standard regimen CA in advanced-stage HD. PMID- 14760123 TI - Intensive chemotherapy and autologous stem-cell transplantation plus rituximab is superior to conventional chemotherapy for newly diagnosed advanced stage mantle cell lymphoma: a matched pair analysis. AB - BACKGROUND: The outcome of 20 patients with newly diagnosed mantle-cell lymphoma (MCL) treated on a prospective trial of autologous stem-cell transplantation (ASCT) and rituximab immunotherapy was compared with the outcome of 40 matched historical control patients treated with standard combination chemotherapy. PATIENTS AND METHODS: Control patients with MCL were identified from a lymphoma database, and pairs were matched with patients receiving ASCT-rituximab for stage of disease, gender and age (+/-5 years). Only patients treated with an anthracycline- or cyclophosphamide-fludarabine-based regimen were included. RESULTS: Seventeen of 20 patients who received ASCT-rituximab remain alive in remission at a median of 30 months from diagnosis; one patient relapsed 2 years post-ASCT, and two died at 7 and 11 months post-ASCT without evidence of lymphoma. Of 40 patients treated with conventional chemotherapy, with a median follow-up of 80 months, 33 have relapsed or progressed and 29 have died. Overall (OS) and progression-free (PFS) survival were superior in patients treated with ASCT-rituximab compared with those treated with conventional chemotherapy (PFS at 3 years, 89% versus 29%, P <0.00001; OS at 3 years, 88% versus 65%, P = 0.052). CONCLUSIONS: This matched-pair analysis suggests that patients with advanced stage MCL treated with ASCT-rituximab had statistically significantly better PFS and a trend toward better OS than patients treated with conventional chemotherapy. Longer follow-up will determine response duration and the true impact of this treatment strategy on PFS and OS. PMID- 14760124 TI - Evaluation of the Cockroft-Gault, Jelliffe and Wright formulae in estimating renal function in elderly cancer patients. AB - BACKGROUND: More elderly patients are being treated with chemotherapy. Reliable and accurate measures of renal function are needed to obtain predictable, safe and effective exposure to renally excreted drugs. The Jelliffe, Cockroft-Gault and Wright formulae have been used to evaluate renal function, although they have not been validated in elderly oncology patients. We performed a retrospective evaluation of these formulae using the [51Cr]-ethylenediamine tetraacetic acid ([51Cr]-EDTA) method of measuring glomerular filtration rate (GFR) as the 'gold standard'. PATIENTS AND METHODS: Inclusion criteria were age > or = 70 years and serum creatinine <250 micromol/l, performed within 4 weeks of glomerular filtration rate (GFR) measurement. Creatinine clearance was calculated using the Cockroft-Gault, Jelliffe and Wright formulae. The precision and accuracy of the three formulae were compared with the gold standard. RESULTS: Two hundred and twenty-five patients were evaluated: median age, 74 years (range 70-89); males, 108; females, 117; median creatinine, 84 micromol/l (range 44-186). Correlation coefficients of the Jelliffe, Cockroft-Gault and Wright formulae were similar. In the specific GFR ranges of 50-70, 70-90 and 90-120 ml/min, the bias [mean percentage error (MPE)] was +8%, -4% and -13%, respectively. The degree of bias was greater with the Cockroft-Gault and Jelliffe formulae across the same range of GFR with the MPE being -15%, -25%, -32% and -12%, -19% and -23%, respectively. All three formulae have reduced precision and greater bias at the extremes of GFR. CONCLUSIONS: The Wright formula is the most accurate, precise and least biased formula for the calculation of GFR in elderly patients with a GFR >50 ml/min. These results allow the physician to make a decision regarding the use of the formula based on an expected degree of bias. PMID- 14760125 TI - Use of totally implantable central venous access ports for high-dose chemotherapy and peripheral blood stem cell transplantation: results of a monocentre series of 376 patients. AB - BACKGROUND: The complication rate of central venous totally implantable access ports (TIAP), used for high-dose chemotherapy with autologous stem cell transplantation support, has not been fully investigated to date, due to the almost exclusive use of externalised, tunnelled devices in this clinical setting. PATIENTS AND METHODS: During a 66-month period (from 1 January 1997 to 30 June 2002), 376 patients suffering from breast cancer, ovarian cancer, lymphoma or multiple myeloma were treated with high-dose chemotherapy and autologous stem cell transplantation at the European Institute of Oncology (Milan, Italy). A single type of port was used, constructed from titanium and silicone rubber, connected to a 7.8 F polyurethane catheter (Port-A-Cath; SIMS Deltec, Inc., St Paul, MN, USA) inserted into the subclavian vein. They were followed prospectively for device-related complications until the device was removed, the patient died or the study was closed (30 June 2002). RESULTS: No TIAP-related deaths were observed in this series. Seven pneumothoraxes (1.8%) occurred as a complication of TIAP placement, one patient only (0.2%) requiring a tube thoracostomy. Port pocket infection occurred twice in this series (0.53%, 0.01 episodes/1000 days of use), whereas three patients suffered from port-related bacteraemia (0.8%, 0.016/1000 days of use). Infections were successfully treated with antibiotics; all three cases had the ports removed at programme completion. Four cases of deep vein thrombosis were detected (1.06%, 0.022/1000 days of use); low molecular weight heparin was given, followed by oral anticoagulants. Finally, one case of extravasation occurred (0.26%, 0.005/1000 days of use), requiring port removal and local medical therapy. CONCLUSIONS: The use of TIAPs has resulted in a safe and effective option for high-dose chemotherapy deliverance and stem cell transplantation, in spite of inducing severe neutropenia and increasing the risk of sepsis in this category of oncology patient. PMID- 14760126 TI - Phase II trial of gemcitabine concurrent with radiation for locally advanced squamous cell carcinoma of the head and neck. AB - BACKGROUND: Concurrent chemoradiation is the current standard of treatment for patients with advanced unresectable head and neck squamous cell carcinoma (HNSCC). Due to the potent radiosensitizing properties of gemcitabine, we decided to assess its efficacy and toxicity with concurrent radiation in patients with advanced HNSCC. PATIENTS AND METHODS: From January 1997 to December 2001, 27 patients with locally advanced HNSCC (stage III, 37%; stage IV, 63%) were enrolled. All received a course of radiotherapy (70 Gy over 7 weeks) concurrent with weekly infusions of gemcitabine at 100 mg/m2 or 50 mg/m2. RESULTS: All patients were assessable for toxicity and 26 for response. Severe mucositis (grade 3-4) was observed in 74% of patients (grade 4, 41%). Severe hematological toxicity was uncommon. Mild and moderate xerostomy was the most common late toxicity in 23 patients (85%). The median radiation dose delivered was 70 Gy (40 80 Gy), 25 patients (93%) received > or = 80% of the intended dose. Gemcitabine dose intensity was > or = 80% in only 13 (48%) patients. The rate of complete and partial responses were 61% and 27%, respectively, for an overall response rate of 88%. At a median follow-up of 13 months (range 6-62), the actuarial 3-year progression-free survival (PFS) and overall survival (OS) were 37% and 33%, respectively. The only variable associated with prolonged survival (P = 0.0001) was the degree of response. No difference was observed in response or toxicity with either gemcitabine 50 or 100 mg/m2. CONCLUSIONS: The concurrent use of radiotherapy and gemcitabine is effective but produces manageable severe mucositis in a high percentage of patients. PMID- 14760127 TI - Conformity to clinical practice guidelines, multidisciplinary management and outcome of treatment for soft tissue sarcomas. AB - BACKGROUND: The primary management of adult soft tissue sarcomas (STS) is characterized by heterogeneity across centers. Several studies suggest that it is improved when coordinated by specialized sarcoma centers. PATIENTS AND METHODS: This study, comparing STS patients of the Rhone-Alpes region treated within and outside the cancer network, retrospectively assesses the conformity of medical practice with 'evidence-based medicine' (EBM) reported under the clinical practice guidelines (CPGs) of the French Federation of Cancer Centers. Institutional records of 100 new STS patients seen between 1999 and 2001 in the regional comprehensive cancer center and Lyon University hospital were analyzed retrospectively (50/300 new files randomly selected in each institution). Medical decisions were checked for conformity with CPGs. RESULTS: Median age was 58 years (range 18-88) and median tumor size was 9 cm (range 1-26). The most common primary sites were extremities, viscera or trunk. The most frequent histology was leiomyosarcoma (21%) or liposarcoma (12%). Only 7% of cases were reviewed by formal multidisciplinary committee before biopsy (with 42% pre-surgery biopsies only). The first surgical resection was R0, R1 and R2 in 26, 29 and 45% of cases, respectively. Conformity to CPGs was rated 52, 81, 94 and 95% for initial surgery, radiation therapy, chemotherapy and follow-up, respectively. At multivariate analysis, pre-surgery multidisciplinary discussion, management in reference center and management within cancer network independently predicted conformity to CPGs. CONCLUSIONS: Conformity with EBM was similar to previous reports. Elaboration of treatment strategy within a formal multidisciplinary staff and treatment within a cancer network are both important prognostic factors for optimal clinical care. PMID- 14760128 TI - Phase I study of PKC412 (N-benzoyl-staurosporine), a novel oral protein kinase C inhibitor, combined with gemcitabine and cisplatin in patients with non-small cell lung cancer. AB - BACKGROUND: PKC412 (N-benzoyl-staurosporine), an oral inhibitor of protein kinase C, is capable of cell cycle inhibition and is endowed with anti-angiogenic properties. This dose-finding phase I study was designed to establish the maximum tolerated dose (MTD) of PKC412 when combined with cisplatin-gemcitabine. PATIENTS AND METHODS: Escalating doses of PKC412 were given every day of a 4 week cycle with cisplatin 100 mg/m2 on day 2 and gemcitabine 1000 mg/m2 on days 1, 8 and 15 in patients with non-small-cell lung cancer. Dose escalation was based on a modified continuous reassessment method. RESULTS: Twenty-three patients, assigned to four cohorts receiving PKC412 at a dose ranging from 25 to 150 mg/day were evaluable. Grade 3 diarrhea occurring in 3/4 patients at cycle 1 led us to define 150 mg/day as the MTD. The MTD based on multiple cycles was redefined as 100 mg/day, since prolonged grade 2-3 nausea/vomiting leading to treatment discontinuation occurred in 3/7 patients after repeated cycles. The next lower dose tested of 50 mg/day was therefore considered as the recommended dose for phase II trials. Among 33 cycles in eight patients, toxicity consisted of grade 1 2 diarrhea (12.5%) and asthenia (50%) with only one patient experiencing grade 3 headache at this dose level. A partial response was observed in three patients. CONCLUSIONS: The results of the present study indicate that PKC412 at a dose of 50 mg/day can be safely added to cisplatin and gemcitabine in patients with advanced non-small-cell lung cancer. PMID- 14760129 TI - Phase I clinical trial of a human idiotypic p53 vaccine in patients with advanced malignancy. AB - BACKGROUND: The purpose of this study was to induce immunity to p53 by using an idiotypic vaccine, composed of a pool of eight peptides derived from the complimentarity determining regions (CDRs) of human anti-p53 antibodies. PATIENTS AND METHODS: Subjects with advanced malignancy received up to four, monthly intradermal injections of pooled peptides (500 microg of each) admixed with granulocyte-macrophage colony-stimulating factor (GM-CSF; 100 microg). In addition, two sheep and two rabbits were also vaccinated with the pooled peptides. RESULTS: Fourteen subjects were enrolled into the study and six of these completed the vaccination schedule. The vaccine was well tolerated by all subjects and no major adverse events were attributable to the vaccine. All subjects mounted in vivo delayed type hypersensitivity (DTH) responses to two or more of the individual vaccine peptides. Vaccine-induced antibodies specific for peptides 2, 5 or 8 were detected in four of six subjects, and two of these had vaccine-specific, cell-mediated responses. Increasing titers of p53-specific antibodies were found in one patient. No T-cell response to p53 was observed in any of the subjects. All animals developed humoral immunity to the peptides and one of the sheep developed rising serum titers of anti-p53 antibodies. CONCLUSIONS: Vaccination with human antibody CDR regions represents a novel method for inducing human antibodies, which may in turn serve as immunological mimics of p53. PMID- 14760130 TI - Efficacy, safety and pharmacokinetics of palonosetron in patients receiving highly emetogenic cisplatin-based chemotherapy: a dose-ranging clinical study. AB - BACKGROUND: Although currently available 5-hydroxytryptamine type 3 receptor (5 HT3) antagonists are effective, not all patients receiving these agents achieve adequate control of chemotherapy-induced nausea and vomiting (CINV). Palonosetron, a potent and highly selective 5-HT3 antagonist with a strong affinity for 5-HT3 and a prolonged plasma elimination half-life, may provide a longer duration of action than other approved agents. PATIENTS AND METHODS: One hundred and sixty-one patients were randomly assigned to receive a single intravenous bolus dose of palonosetron (0.3, 1, 3, 10, 30 or 90 microg/kg) before administration of highly emetogenic chemotherapy, with no pretreatment with corticosteroids. RESULTS: The four highest doses of palonosetron were similarly effective during the first 24 h, producing clearly higher complete response (CR) (no emesis, no rescue medication) rates in the 3, 10, 30 and 90 microg/kg groups (46%, 40%, 50% and 46%, respectively) than in the 0.3-1 microg/kg group (24%) of evaluable patients (n = 148). The 3 microg/kg dose was identified as the lowest effective dose. A single dose of palonosetron showed prolonged efficacy in preventing delayed emesis, with approximately one-third of patients who received palonosetron 10 or 30 microg/kg maintaining a CR throughout the 7-day period following chemotherapy administration. Dose-proportional increases in pharmacokinetic parameters and a long plasma half-life (43.7-128 h) were observed. Palonosetron was well-tolerated, with no dose-response effect evident for the incidence or intensity of adverse events. CONCLUSIONS: Palonosetron is an effective and well-tolerated agent for the prevention of CINV following highly emetogenic chemotherapy, with 3 and 10 microg/kg identified as the lowest effective palonosetron doses. PMID- 14760131 TI - Monitoring falls in gastric cancer mortality in Europe. AB - We have considered trends in age-standardized mortality from gastric cancer in 25 individual European countries, as well as in the European Union (EU) as a whole, in six selected central-eastern European countries and in the Russian Federation over the period 1950-1999. Steady and persisting falls in rates were observed, and the fall between 1980 and 1999 was approximately 50% in the EU, 45% in eastern Europe and 40% in Russia. However, the declines were greater in Russia and eastern Europe, since rates were much higher, in absolute terms. Joinpoint regression analysis indicated that the falls were proportionally greater in the last decade for men (-3.83% per year in the EU) and in the last 25 years for women (-3.67% per year in the EU) than in previous calendar years. Moreover, steady declines in gastric cancer mortality were observed in the middle-aged and the young population as well, suggesting that they are likely to persist in the near future. In terms of number of deaths avoided, however, the impact of the decline in gastric cancer mortality will be smaller, particularly in the EU. PMID- 14760132 TI - Processed meat and the risk of selected digestive tract and laryngeal neoplasms in Switzerland. AB - BACKGROUND: Processed meat has been related to the risk of digestive tract neoplasms but the evidence remains inconclusive. We examined data from a network of case-control studies conducted between 1992 and 2002 in the Swiss Canton of Vaud. PATIENTS AND METHODS: We studied 316 patients with incident, histologically confirmed oral and pharyngeal cancer, 138 patients with oesophageal cancer, 91 patients with laryngeal cancer and 323 patients with colorectal cancer. Controls were 1271 subjects admitted to the same hospital for a wide spectrum of acute non neoplastic conditions, unrelated to long-term modification of diet. RESULTS: There were strong direct trends in risk between consumption of processed meat and the various neoplasms considered: the multivariate odds ratios for the highest quartile of intake compared to the lowest were 4.7 for oral and pharyngeal cancer, 4.5 for oesophageal cancer, 3.4 for laryngeal cancer and 2.5 for colorectal cancer. The association was stronger in younger subjects, in moderate drinkers and in non-smokers. CONCLUSION: Processed meat represents a strong indicator of unfavourable diet for digestive tract and laryngeal cancer risk in this population. PMID- 14760133 TI - Set-up of a population-based familial breast cancer registry in Geneva, Switzerland: validation of first results. AB - BACKGROUND: This article evaluates the accuracy of family history of breast and ovarian cancer among first-degree relatives of breast cancer patients, retrospectively collected during the setting up of a population-based family breast cancer registry. PATIENTS AND METHODS: Family histories of cancer for all women with breast cancer recorded at the Geneva Cancer Registry from 1990 to 1999 were retrospectively extracted from medical files. The accuracy of these family histories was validated among Swiss women born in Geneva: all 119 with a family history of breast (n = 110) or ovarian (n = 9) cancer and a representative sample of 100 women with no family history of breast or ovarian cancer. We identified the first-degree relatives of these women with information from the Cantonal Population Office. All first-degree relatives, resident in Geneva from 1970 to 1999, were linked to the cancer registry database for breast and ovarian cancer occurrence. Sensitivity, specificity and level of overall agreement (kappa) were calculated. RESULTS: Among 310 first-degree relatives identified, 61 had breast cancer and six had ovarian cancer recorded at the Geneva Cancer Registry. The sensitivity, specificity and kappa of the reported family histories of breast cancer were 98%, 97% and 0.97, respectively. For ovarian cancer, the sensitivity, specificity and kappa were 67%, 99%, and 0.66, respectively. CONCLUSIONS: This study indicates that retrospectively obtained family histories are very accurate for breast cancer. For ovarian cancer, family histories are less precise and may need additional verification. PMID- 14760134 TI - Lesson learned from high-dose chemotherapy for high-risk breast cancer (What you see is what you mean). PMID- 14760135 TI - Successful treatment of AIDS-related Castleman's disease following the administration of highly active antiretroviral therapy (HAART). PMID- 14760136 TI - Postoperative chemoradiotherapy after curative gastrectomy for cancer. PMID- 14760137 TI - Pigmentary changes in chronic myeloid leukemia patients treated with imatinib mesylate. PMID- 14760138 TI - First-line hepatic infusion of pirarubicin in patients with isolated liver metastases: is it really promising? PMID- 14760139 TI - Written asthma action plans. PMID- 14760140 TI - COPD in the ECRHS. PMID- 14760141 TI - New insights into oxygen sensing at a cellular level. PMID- 14760142 TI - Alpha1-antitrypsin deficiency. PMID- 14760143 TI - Written action plans for asthma: an evidence-based review of the key components. AB - BACKGROUND: Written action plans for asthma facilitate the early detection and treatment of an asthma exacerbation. Several versions of action plans have been published but the key components have not been determined. A study was undertaken to determine the impact of individual components of written action plans on asthma health outcomes. METHODS: Randomised controlled trials (n=26) that evaluated asthma action plans as part of asthma self-management education were identified. Action plans were classified as being individualised and complete if they specified when and how to increase treatment (n=17), and as incomplete (n=4) or non-specific (n=5) if they did not include these instructions. RESULTS: For individualised complete written action plans the use of 2-4 action points and the use of both inhaled (ICS) and oral (OCS) corticosteroid consistently improved asthma outcomes. Action points based on personal best peak expiratory flow (PEF) consistently improved health outcomes while those based on percentage predicted PEF did not. The efficacy of incomplete action plans was inconclusive because of insufficient data. Non-specific action plans led to improvements in knowledge and symptoms. CONCLUSION: Individualised written action plans based on personal best PEF, using 2-4 action points, and recommending both ICS and OCS for treatment of exacerbations consistently improve asthma health outcomes. Other variations appear less beneficial or require further study. These observations provide a guide to the types of variations possible with written action plans, and strongly support the use of individualised complete written action plans. PMID- 14760145 TI - Increase in urinary leukotriene LTE4 levels in acute asthma: correlation with airflow limitation. AB - BACKGROUND: Leukotrienes play a key role in the pathophysiology of chronic asthma. Activation of leukotriene pathways is accompanied by rises in detectable urinary levels of leukotriene E4 (LTE4). The relationship between urinary LTE4 levels and factors associated with acute asthma has not been determined. METHODS: Adults aged 15-54 years presenting with moderate to severe acute asthma were evaluated at emergency departments in 16 US sites. Forced expiratory volume in 1 second (FEV1) was measured during the first 60 minutes after arrival and at specified times until discharge or admission. Urine samples for measurement of LTE4 levels were obtained either on arrival at the study site and/or before discharge. Patients were seen 2 weeks later for follow up, at which time repeat FEV1 measurements and urine samples for LTE4 were obtained. RESULTS: One hundred and eighty four patients were evaluated; LTE4 results from both the acute and follow up periods were available for analysis in 146. Urinary LTE4 levels were increased during asthma exacerbations compared with levels obtained 2 weeks later (geometric means 111.7 and 75.6 pg/mg creatinine, respectively, mean percentage change -32.3; 95% confidence interval (CI) for the mean percentage change -39.6 to -24.3, p<0.001). The correlation between improvement in FEV1 and decline in LTE4 over the 2 week interval was significant (p<0.001, r=0.43). CONCLUSIONS: Activation of leukotriene pathways in acute asthma is correlated with the degree of airflow obstruction, and resolution of the asthma exacerbation is associated with a reduction in leukotriene levels. PMID- 14760148 TI - Nebulised salbutamol administered during sputum induction improves bronchoprotection in patients with asthma. AB - BACKGROUND: Inhalation of hypertonic or even isotonic saline during sputum induction may cause bronchospasm in susceptible patients with asthma, despite premedication with 400 microg inhaled salbutamol delivered by pressurised metered dose inhaler (pMDI). The bronchoprotection afforded by additional inhaled salbutamol administered through the ultrasonic nebuliser during sputum induction was investigated. METHODS: Twenty patients with moderate to severe asthma underwent sputum induction by inhaling saline 4.5% (or 0.9% if post bronchodilation forced expiratory volume in 1 second (FEV1) <65% predicted) for 10 minutes according to two protocols given 1 week apart in random order. At visit A the patients received 400 microg salbutamol administered through a pMDI+spacer 20 minutes before induction while at visit B the premedication was supplemented by 1500 microg nebulised salbutamol inhaled throughout the induction procedure. Both the investigator and the patients were blind to the nebulised solution used. FEV1 was recorded during sputum induction at 1, 3, 5, and 10 minutes. Sputum cell counts and histamine, tryptase and albumin levels in the supernatants were determined. RESULTS: The mean (SE) maximal reduction in FEV1 over the 10 minute period of sputum induction was 11.7 (2.8)% at visit A, which was significantly greater than at visit B (2.6 (1.2)%; mean difference 9% (95% CI 2.7 to 15.4), p<0.01). Total and differential sputum cell counts as well as albumin, tryptase, and histamine levels did not differ between the two visits. CONCLUSION: The addition of inhaled salbutamol through an ultrasonic nebuliser markedly improves bronchoprotection against saline induced bronchoconstriction in patients with moderate to severe asthma undergoing sputum induction without affecting cell counts and inflammatory markers. PMID- 14760147 TI - Fatty acid levels and risk of asthma in young adults. AB - BACKGROUND: There is current interest in the possible protective effect of long chain (n-3) fatty acids from fish in chronic lung diseases such as asthma. The aim of this community based cross sectional study was to determine whether plasma long chain (n-3) fatty acids, as a measure of dietary intake, differed between 1601 young adults with and without asthma. METHODS: Subjects of mean (SD) age 34.6 (7.1) years completed a detailed respiratory questionnaire, food frequency questionnaire, skin prick testing, and lung function tests including methacholine challenge test for bronchial hyperreactivity (BHR) and had venous blood taken for analysis of plasma fatty acids. Plasma fatty acid levels (%) were analysed using multiple logistic regression with alternative definitions of asthma and atopy as the outcomes. RESULTS: Atopy was not associated with any plasma fatty acid. The n 3 polyunsaturated fatty acids and n-6:n-3 ratio were not consistently associated with asthma or atopy. The n-6 polyunsaturated fatty acid dihomo gamma-linolenic acid (DHGLA) was positively associated with current asthma (OR=1.30, 95% CI 1.06 to 1.60), asthma (OR=1.34, 95% CI 1.13 to 1.60), and doctor diagnosed asthma (OR=1.25, 95% CI 1.06 to 1.48). CONCLUSION: Plasma n-3 fatty acids are not associated with a reduced risk of asthma or atopy among young adults. The association of DHGLA with asthma warrants further research to determine a cause effect relationship. PMID- 14760150 TI - Two functional variants of the superoxide dismutase genes in Finnish families with asthma. AB - BACKGROUND: Functional polymorphisms in the genes encoding superoxide dismutases (SOD)-that is, superoxide scavenging antioxidant enzymes-may play an important role in the development of inflammatory airway diseases such as asthma. METHODS: The allele frequencies of two missense polymorphisms of SOD genes (Ala16Val in MnSOD (SOD2) and Arg213Gly in ECSOD (SOD3)) were investigated in Finnish patients with asthma and compared with family based controls. Both variants have been shown to be functionally interesting in the lung. The polymorphism at the exon intron 3 boundary of a third SOD, CuZnSOD (SOD1), was also included in the analysis. RESULTS: None of the SOD genetic variants studied appeared to be major genetic regulators in the development of asthma. We could exclude all models of inheritance that increased the risk of asthma more than 1.2 fold for MnSOD*Val (frequency of allele 0.74 in the population) and more than 6.6 fold for ECSOD*Gly213 (frequency of allele 0.03 in the population) compared with non carriers. For the intronic polymorphism in CuZnSOD, a relative risk of more than 3.3 (frequency of allele 0.10 in the population) could be excluded. CONCLUSIONS: It is highly unlikely that the functionally important genetic variants Ala16Val and Arg213Gly of SODs play a major role in the genetic susceptibility of asthma. PMID- 14760151 TI - An international survey of chronic obstructive pulmonary disease in young adults according to GOLD stages. AB - BACKGROUND: The recently published GOLD guidelines provide a new system for staging chronic obstructive pulmonary disease (COPD) from mild (stage I) to very severe (stage IV) and introduce a stage 0 (chronic cough and phlegm without airflow obstruction) that includes subjects "at risk" of developing the disease. METHODS: In order to assess the prevalence of GOLD stages of COPD in high income countries and to evaluate their association with the known risk factors for airflow obstruction, data from the European Community Respiratory Health Survey on more than 18,000 young adults (20-44 years) were analysed. RESULTS: The overall prevalence was 11.8% (95% CI 11.3 to 12.3) for stage 0, 2.5% (95% CI 2.2 to 2.7) for stage I, and 1.1% (95% CI 1.0 to 1.3) for stages II-III. Moderate to heavy smoking (> or =15 pack years) was significantly associated with both stage 0 (relative risk ratio (RRR)=4.15; 95% CI 3.55 to 4.84) and stages I+ (RRR=4.09; 95% CI 3.17 to 5.26), while subjects with stages I+ COPD had a higher likelihood of giving up smoking (RRR=1.39; 95% CI 1.04 to 1.86) than those with GOLD stage 0 (RRR=1.05; 95% CI 0.86 to 1.27). Environmental tobacco smoke had the same degree of positive association in both groups. Respiratory infections in childhood and low socioeconomic class were significantly and homogeneously associated with both groups, whereas occupational exposure was significantly associated only with stage 0. All the GOLD stages showed a significantly higher percentage of healthcare resource users than healthy subjects (p<0.001), with no difference between stage 0 and COPD. CONCLUSIONS: A considerable percentage of young adults already suffered from COPD. GOLD stage 0 was characterised by the presence of the same risk factors as COPD and by the same high demand for medical assistance. PMID- 14760152 TI - Transforming growth factor-beta1 genotype and susceptibility to chronic obstructive pulmonary disease. AB - BACKGROUND: Only a few long term smokers develop symptomatic chronic obstructive pulmonary disease (COPD) and this may be due, at least in part, to genetic susceptibility to the disease. Transforming growth factor beta1 (TGF-beta1) has a number of actions that make it a candidate for a role in the pathogenesis of COPD. We have investigated a single nucleotide polymorphism at exon 1 nucleotide position 29 (T-->C) of the TGF-beta1 gene that produces a substitution at codon 10 (Leu-->Pro). METHODS: The frequency of this polymorphism was determined in 165 subjects with COPD, 140 healthy blood donors, and 76 smokers with normal lung function (resistant smokers) using the polymerase chain reaction and restriction enzyme fragment length polymorphism. RESULTS: The distribution of genotypes was Leu-Leu (41.8%), Leu-Pro (50.3%), and Pro-Pro (7.9%) for subjects with COPD, which was significantly different from the control subjects (blood donors: Leu Leu (29.3%), Leu-Pro (52.1%) and Pro-Pro (18.6%), p=0.006; resistant smokers: Leu Leu (28.9%), Leu-Pro (51.3%) and Pro-Pro (19.7%), p=0.02). The Pro10 allele was less common in subjects with COPD (33%) than in blood donors (45%; OR=0.62, 95% CI 0.45 to 0.86, p=0.005) and resistant smokers (45%; OR=0.59, 95% CI 0.40 to 0.88, p=0.01). CONCLUSIONS: The proline allele at codon 10 of the TGF-beta1 gene occurs more commonly in control subjects than in individuals with COPD. This allele is associated with increased production of TGF-beta1 which raises the possibility that TGF-beta1 has a protective role in COPD. PMID- 14760153 TI - Local activation of coagulation and inhibition of fibrinolysis in the lung during ventilator associated pneumonia. AB - BACKGROUND: Fibrin deposition is a hallmark of pneumonia. To determine the kinetics of alterations in local coagulation and fibrinolysis in relation to ventilator associated pneumonia (VAP), a single centre prospective study of serial changes in pulmonary and systemic thrombin generation and fibrinolytic activity was conducted in patients at risk for VAP. METHODS: Non-directed bronchial lavage (NBL) was performed on alternate days in patients expected to require mechanical ventilation for more than 5 days. A total of 28 patients were studied, nine of whom developed VAP. RESULTS: In patients who developed VAP a significant increase in thrombin generation was observed in the airways, as reflected by a rise in the levels of thrombin-antithrombin complexes in NBL fluid accompanied by increases in soluble tissue factor and factor VIIa concentrations. The diagnosis of VAP was preceded by a decrease in fibrinolytic activity in NBL fluid. Indeed, before VAP was diagnosed clinically, plasminogen activator activity levels in NBL fluid gradually declined, which appeared to be caused by a sharp increase in NBL fluid levels of plasminogen activator inhibitor 1. CONCLUSION: VAP is characterised by a shift in the local haemostatic balance to the procoagulant side, which precedes the clinical diagnosis of VAP. PMID- 14760154 TI - Rapid and efficient clearance of airway tissue granulocytes through transepithelial migration. AB - BACKGROUND: Clearance of tissue granulocytes is central to the resolution of airway inflammation. To date the focus has been on apoptotic mechanisms of cell removal and little attention has been given to alternative processes. The present study explores transepithelial migration as a mechanism of cell clearance. METHOD: Guinea pig tracheobronchial airways where eosinophils are constitutively present in the mucosal tissue were studied. A complex topical stimulus (allergen challenge) was applied and the fate of the eosinophils was determined by selective tracheobronchial lavage and histological examination of the tissue. RESULTS: Within 10 minutes of the allergen challenge, massive migration of eosinophils into the airway lumen occurred together with a reduction in tissue eosinophil numbers. Cell clearance into the lumen continued at high speed and by 30 and 60 minutes the tissue eosinophilia had been reduced by 63% and 73%, respectively. The marked transepithelial migration (estimated maximal speed 35,000 cells/min x cm2 mucosal surface) took place ubiquitously between epithelial cells without affecting epithelial integrity as assessed by transmission and scanning electron microscopy. Eosinophil apoptosis was not detected but occasional cytolytic eosinophils occurred. CONCLUSION: This study shows that luminal entry has a remarkably high capacity as a granulocyte elimination process. The data also suggest that an appropriate stimulus of transepithelial migration may be used therapeutically to increase the resolution of inflammatory conditions of airway tissues. PMID- 14760156 TI - BTS randomised feasibility study of active symptom control with or without chemotherapy in malignant pleural mesothelioma: ISRCTN 54469112. AB - BACKGROUND: The incidence of mesothelioma is rising rapidly in the UK. There is no generally accepted standard treatment. The BTS recommends active symptom control (ASC). It is not known whether chemotherapy in addition prolongs survival or provides worthwhile palliation with acceptable toxicity. Palliation as recorded by patients has been fully reported for only two regimens: mitomycin, vinblastine, and cisplatin (MVP), and vinorelbine (N). The BTS and collaborators planned to conduct a phase III randomised trial comparing ASC only, ASC+MVP, and ASC+N in 840 patients with survival as the primary outcome measure. The aim of the present study was to assess the acceptability of the trial design to patients and the suitability of two standard quality of life (QL) questionnaires for mesothelioma. METHODS: Collaborating centres registered all new patients with mesothelioma. Those eligible and giving informed consent completed EORTC QLQ C30+LC13 and FACT-L QL questionnaires and were randomised between all three or any two of (1) ASC only, (2) ASC+4 cycles of MVP, and (3) ASC+12 weekly doses of N. RESULTS: During 1 year, 242 patients were registered of whom 109 (45%) were randomised (55% of the 197 eligible patients). Fifty two patients from 20 centres were randomised to an option including ASC only. This translates into a rate of 312 per year from 60 centres interested in collaborating in the phase III trial. The EORTC QL questionnaire was superior to FACT-L in terms of completeness of data and patient preference. Clinically relevant palliation was achieved with ASC. CONCLUSION: The planned phase III trial is feasible. PMID- 14760157 TI - Bone mineral density in Australian children, adolescents and adults with cystic fibrosis: a controlled cross sectional study. AB - BACKGROUND: Low bone mineral density (BMD) is recognised in individuals with cystic fibrosis (CF) although the pathogenesis remains unclear. The aims of this study were to compare BMD over a broad continuum of Australian individuals with CF with healthy controls and to examine the relationship between BMD and clinical parameters including physical activity, nutrition, and vitamin D levels. METHODS: BMD of the lumbar spine (LS), total body (TB), femoral neck (FN), cortical wrist (R33%), and distal wrist (RUD) was examined in 153 individuals with CF aged 5.3 55.8 years (84 males) and in 149 local controls aged 5.6-48.3 years (66 males) using dual energy x ray absorptiometry. Anthropometric variables, body cell mass, markers of disease severity, corticosteroid usage, measures of physical activity, dietary calcium and caloric intake and serum vitamin D were assessed and related to BMD. RESULTS: Compared with controls, mean BMD was not significantly different in children aged 5-10 years with CF. Adolescents (females 11-18 years, males 11 20 years) had reduced TB and R33% BMD when adjusted for age, sex, and height (difference in BMD (g/cm2) adjusted means between control and CF: TB=0.04 (95% CI 0.01 to 0.07); R33%=0.03 (95% CI 0.01 to 0.06)). BMD was reduced at all sites except R33% in adults (difference in BMD (g/cm2) adjusted means between control and CF: TB=0.05 (95% CI 0.02 to 0.09); LS=0.08 (95% CI 0.03 to 0.14); FN=0.09 (95% CI 0.03 to 0.15); RUD=0.03 (95% CI 0.01 to 0.05)). In children/adolescents BMD was weakly associated with nutritional status and disease severity. CONCLUSIONS: BMD was normal in a well nourished group of prepubertal children with CF. A BMD deficit appears to evolve during adolescence and becomes more marked in adults. Individuals with CF should optimise nutrition, partake in physical activity, and maximise lung health in order to optimise BMD. Further longitudinal studies are required to understand the evolution of reduced BMD in young people and adults with CF. PMID- 14760158 TI - Nitric oxide synthase 1 as a potential modifier gene of decline in lung function in patients with cystic fibrosis. AB - BACKGROUND: The severity of lung disease varies widely in patients with cystic fibrosis (CF) who have the same type of mutations of the cystic fibrosis transmembrane regulator (CFTR) gene, suggesting involvement of "modifier" genes. The nitric oxide synthase 1 (NOS1) gene is a candidate for this role because exhaled nitric oxide (NO) is reduced in patients with CF and NOS1 activity contributes to transepithelial ionic transport, immune defence, and non-specific inflammation of the airways. METHODS: Dinucleotide GT repeat polymorphism was studied in the 5' untranslated region of the NOS1 gene, immediately upstream from the transcription initiation site, in 59 patients with CF and 59 healthy controls. RESULTS: Nineteen alleles of the NOS1 gene were identified according to the number of GT repeats (from 18 to 36) in the 5 untranslated region. Exhaled NO levels were significantly correlated with the number of GT repeats. Patients with CF who had the NOS1 genotype associated with high NO production had a slower decline in lung function during the 5 year follow up period. There was no confounding effect of age, chronic bacterial colonisation of the airway, or CFTR genotype. CONCLUSIONS: These data suggest a possible link between the NOS1 gene locus and the rate of decline in lung function in patients with CF. PMID- 14760159 TI - Sleep. 2: pathophysiology of obstructive sleep apnoea/hypopnoea syndrome. AB - The pathogenesis of airway obstruction in patients with obstructive sleep apnoea/hypopnoea syndrome is reviewed. The primary defect is probably an anatomically small or collapsible pharyngeal airway, in combination with a sleep induced fall in upper airway muscle activity. PMID- 14760160 TI - Alpha1-antitrypsin deficiency. 1: epidemiology of alpha1-antitrypsin deficiency. AB - The protein and molecular characteristics of variants of the alpha1-antitrypsin (AAT) gene are described, and available data on the genetic epidemiology of AAT deficiency are presented. PMID- 14760162 TI - Elimination of central sleep apnoea by mitral valvuloplasty: the role of feedback delay in periodic breathing. AB - Central sleep apnoea is a form of periodic breathing which resembles Cheyne Stokes respiration but occurs only during sleep. One mechanism in the pathogenesis is a delay in chemical feedback from the lungs to the medullary respiratory centre. We explored the relationship between circulatory feedback delay in a patient with central sleep apnoea and Cheyne-Stokes respiration before and after mitral valve repair. Preoperatively the patient had severe central sleep apnoea and an increased circulation time. Following mitral valvuloplasty the circulation time was decreased with resolution of central sleep apnoea. This case demonstrates the role of feedback delay in central sleep apnoea and suggests that similar haemodynamic mechanisms may lead to central sleep apnoea and Cheyne Stokes respiration. PMID- 14760161 TI - Oxidants and asthma. AB - Many decades of research have produced a significant amount of data showing increased oxidative stress in asthma and indicating a potential role for oxidants in the pathogenesis of the disease, particularly during exacerbations. Putatively relevant pro-oxidative mechanisms have also been identified. Currently available asthma drugs are generally effective for the treatment of the disease, but their effects on oxidative stress have still not been completely elucidated. From the data available in the literature one can conclude that antioxidant compounds may have a potential role in the treatment of asthma, especially of asthma exacerbations. More convincing evidence from controlled clinical trials is required. PMID- 14760163 TI - Squawks in pneumonia. PMID- 14760164 TI - BTS guidelines for the management of pleural infection. PMID- 14760165 TI - Bronchodilator reversibility testing in COPD. PMID- 14760166 TI - Prednisolone response in patients with COPD. PMID- 14760167 TI - Breathing exercises in asthma. PMID- 14760168 TI - Selective pulmonary fat aspiration complicating oesophageal achalasia. PMID- 14760169 TI - Intravenous therapy. AB - Intravenous administration of fluids, drugs, and nutrition is very common in hospitals. Although insertion of peripheral and central cannulae and subsequent intravenous therapy are usually well tolerated, complications that prolong hospitalisation, and in some cases cause death, can arise on occasions. Additionally, many cannulae are inserted unnecessarily. This article seeks to review this area and to outline good medical practice. PMID- 14760170 TI - How, and when, can I restrain a patient? AB - The need to restrain service users will vary according to the area of practice within which practitioners are employed. Many of the principles that relate to the issue of restraining service users are, however, applicable to all, or most, health and social care settings. While the emphasis should be on pre-emptive action, wherever possible, in order to prevent the need to restrain, there are some occasions on which the risks to the service user, or others, of inaction may outweigh those of taking action. Some of the key issues are discussed in this paper. PMID- 14760172 TI - Suture granuloma. PMID- 14760171 TI - Drug-eluting stents: new era and new concerns. AB - At present there is much excitement about drug-eluting stents, which hold promise for the treatment of coronary artery disease. This ingenious therapy involves coating the outside of a standard coronary stent with a thin polymer containing medication that can prevent scarring at the site of coronary intervention. Early trials with sirolimus coated stents showed that they might prevent coronary artery restenosis, but later studies, involving more complex coronary lesions, did not show a complete absence of restenosis. Recent studies have demonstrated the long term cost effectiveness of drug-eluting stents as they have reduced the need for revascularisation procedures. At present there are few data on the safety and effectiveness of stents over follow up periods exceeding two years, and data obtained from animal models of stenting might not be completely applicable to humans. There are concerns that drug-eluting stents might delay, rather than inhibit, restenosis. Also there is concern regarding the inflammation caused by the polymer substrate. This article reviews the present data on drug eluting stents and their benefits, shortcomings, and concerns. PMID- 14760173 TI - Cardiac magnetic resonance imaging. AB - Recent advances in technology have allowed magnetic resonance imaging to be exploited in cardiology. It is capable of giving high definition structural and functional information. The current applications of this technique and how it may develop in the future are reviewed. PMID- 14760174 TI - Resource utilisation, length of hospital stay, and pattern of investigation during acute medical hospital admission. AB - OBJECTIVES: To describe the patient demographic characteristics and organisational factors that influence length of stay (LOS) among emergency medical admissions. Also, to describe differences in investigation practice among consultant physicians and to examine the impact of these on LOS. DESIGN: Prospective observational study. SETTING: General medicine department of a teaching hospital in Belfast, UK. PARTICIPANTS: Data were recorded for patients who were admitted as emergencies and reviewed on the post-take ward rounds (PTWR) attended by the investigation coordinator. OUTCOME MEASURES: Non-laboratory investigations requested, LOS, and diagnosis on discharge. RESULTS: Of 830 episodes evaluated, the median LOS was 7 days (interquartile range 3-12 days); this was significantly longer for admissions on Fridays (p = 0.0011) and for patients managed on medical wards (p<0.0001). There was a positive correlation between patient age and LOS (r = 0.32, p<0.0001). Chest radiographs (p = 0.002) and echocardiography (p = 0.015) were associated with a prolonged LOS; no investigations were associated with a shortened LOS. Diagnoses of congestive heart failure, respiratory disease, and cancer were associated with a longer LOS; a diagnosis of angina was associated with a shorter LOS. Considerable variation in investigation ordering, but no difference in LOS, was observed between consultants. High use of a given medical test did not correlate with high use of other tests. CONCLUSION: A systematic means of dealing with the NHS resource crisis should include an improved organisational strategy as well as social care provision. A more unified approach to investigation practice should also have a sparing effect on resources. PMID- 14760175 TI - Severe herpes zoster after infliximab infusion. PMID- 14760176 TI - How well do we investigate patients with suspected subarachnoid haemorrhage? The continuing need for cerebrospinal fluid investigations. AB - OBJECTIVE: To demonstrate the extent of compliance with established guidelines for the investigation of suspected subarachnoid haemorrhage (SAH) and the implications of non-compliance. DESIGN: Prospective observational study of practice in three hospitals in the Trent region. SETTING: One teaching hospital with a tertiary neuroscience referral centre and two large district general hospitals. PARTICIPANTS: 50 consecutive patients from each centre referred for suspected SAH with negative computed tomography. MAIN OUTCOME MEASURES: Diagnosis of SAH confirmed or excluded according to guidelines. RESULTS: When the data from the three centres were combined (n = 150 computed tomography negative cases) cerebrospinal fluid (CSF) investigation was not performed in 60/150 (40%). In the 90 cases where CSF studies were performed SAH was confirmed in 11 (12%). CONCLUSION: There is significant non-compliance in following the established guidelines for the investigation of SAH at the centres studied. As the primary cause of non-traumatic SAH is ruptured aneurysm, which is associated with high morbidity and mortality from second haemorrhage, this highlights a major source of concern for clinical governance. PMID- 14760177 TI - Attitudes of preclinical medical students towards caring for chronically ill and dying patients: does palliative care teaching make a difference? AB - INTRODUCTION: Students entering medical school today will encounter an ageing population and a higher incidence of diseases affecting the elderly-for example, chronic respiratory and cardiac disease and malignancy. PURPOSE: This study was carried out to determine the attitudes of preclinical medical students towards the care of patients for whom a cure is not possible. METHODS: All students were invited to complete a 23 item questionnaire prior to initial teaching and again following the second teaching session in palliative care. RESULTS: Overall, 149 of the 186 students (80%) completed the pre-teaching questionnaire (59 males and 90 females; median age 20 years, range 19-27 years), and 66 students (35%) completed the post-teaching questionnaire. Attitudes towards chronically ill and dying patients were generally positive. It was found that increasing age was associated with a more positive view of caring for patients with chronic or terminal illness, a more positive view of listening to patients reminisce, and a more positive view of patients dying at home (p = 0.014). The only notable result was that after palliative care teaching students had a significantly more positive view of hospices. CONCLUSION: Caring for patients at the end of life can be one of the most rewarding aspects of being a doctor. This study suggests that the majority of medical students have a positive attitude towards patients with chronic incurable illness, and the trend for encouraging older students to enter medicine may be an influencing factor. PMID- 14760178 TI - Diabetes in a high secure hospital. AB - AIMS: To determine the prevalence of diabetes mellitus and its possible causes and to assess its control in a high secure hospital. METHODS: A cross sectional survey and a prospective cohort study were conducted. The cross sectional survey included 408 patients admitted under the Mental Health Act, and the prospective study included 22 patients with known diabetes followed up for 24 months. The outcome measures evaluated were drug treatment, status of microvascular and macrovascular complications, glycated haemoglobin, and body mass index. RESULTS: In the cross sectional survey, 35 out of 408 patients (8.6%; 95% confidence interval 5.9% to 11.3%) had known diabetes, and all of these had type 2 diabetes. Obesity, cigarette smoking, schizophrenia, and antipsychotic drug use were frequent, and weight gain was common after hospital admission. Glycaemic control was variable, and, although a majority of patients were above recommended treatment targets, control remained stable over the follow up period. CONCLUSIONS: Type 2 diabetes was common in this hospital. Both its prevalence and the suboptimal glycaemic control in some patients probably relate to sedentary life, dietary factors, smoking, and perhaps widespread use of antipsychotic drugs. However, regular multidisciplinary input enabled most patients to maintain relatively stable glycaemic control, with good control of blood pressure and lipids, at levels similar to those seen in community and hospital diabetic clinics. Further modification of lifestyle risk factors is probably needed to reduce the prevalence and impact of diabetes in this patient group. PMID- 14760179 TI - Audit on flexible sigmoidoscopy for rectal bleeding in a district general hospital: are we over-loading the resources? AB - OBJECTIVE: Patients with rectal bleeding are being over investigated because of the fear of missing colorectal cancers. This study aimed to identify the percentage of patients <45 years of age who undergo flexible sigmoidoscopy for rectal bleeding, and to assess and compare the incidence of colorectal cancers and polyps above and below this age. METHODS: Patients who underwent flexible sigmoidoscopy for rectal bleeding between 1 January 2000 and 31 December 2002 were reviewed. Patients were divided into two groups: group 1 consisted of patients aged >or=45 years and group 2 patients <45 years. The histopathology of biopsy specimens taken was also studied. RESULTS: Altogether 18.9% of the patients who had flexible sigmoidoscopy for rectal bleeding were <45 years. The incidence of colorectal cancers in group 1 was 3.5%; all these cases were confirmed on histopathology. Only one patient in group 2 was diagnosed with colorectal cancer on flexible sigmoidoscopy, but the histopathology disproved it. The incidence of polyps was 16.6% in group 1 and 7.9% in group 2. Following histopathology, the incidence of adenomatous polyps was 6.8% in group 1 and 2.1% in group 2. There was a significant difference between the two groups, with a p value of <0.0001. CONCLUSION: The incidence of colorectal cancers and adenomatous polyps in patients aged <45 years with rectal bleeding is very low. A flexible sigmoidoscopy costs approximately pound 330. If new guidelines are implemented considering the age of the patient, considerable cost savings could be made, and the available resources could be appropriately used in groups with high incidences of colorectal cancers. PMID- 14760180 TI - Sphenoidal sinus mucocoele presenting as mono-ocular painless loss of vision. PMID- 14760181 TI - The worshipful Society of Apothecaries of London. AB - The Society of Apothecaries is both a City livery company and an examining authority for the medical profession. Founded in 1617 by the royal apothecary Gideon de Laune leading a breakaway group from the Grocers' Company, the Society was instrumental in raising the status of apothecaries as general practitioners. Under the Apothecaries' Act (1815) the Society examined for the LSA and it now awards the LMSSA (Licence in Medicine and Surgery of the Society of Apothecaries) and postgraduate diplomas, while maintaining the civic, charitable, and ceremonial traditions of a livery company of the City of London. PMID- 14760182 TI - Postcystoscopy confusion. PMID- 14760183 TI - A 67 year old woman with renal failure and sinus bradycardia. PMID- 14760189 TI - Role of enoxaparin in the invasive management of the ACS patient. PMID- 14760188 TI - The combination of enoxaparin, glycoprotein IIb/IIIa inhibitors and an early invasive approach among acute coronary syndrome patients. AB - OBJECTIVES: This study was designed to assess the feasibility and safety of enoxaparin in combination with glycoprotein (GP) IIb/IIIa inhibitors during percutaneous coronary intervention (PCI) as part of an early invasive strategy in patients presenting with acute coronary syndromes (ACS). BACKGROUND: Trials in patients with ACS have evaluated the utility of enoxaparin, adjuvant GP IIb/IIIa inhibitors with PCI, and an early invasive approach. Information about the combination of all three of these approaches, however, is limited. METHODS: Forty nine patients with ACS underwent cardiac catheterization, of whom 23 underwent PCI with enoxaparin and GP IIb/IIIa inhibitors. RESULTS: The primary endpoint of the study, a composite of death, myocardial infarction or urgent revascularization at 30 days, occurred in 8% of patients undergoing PCI. There were no deaths. One patient received a blood transfusion. No other adverse events occurred. These event rates were comparable to those from the pooled EPILOG/EPISTENT database, in which intravenous unfractionated heparin was used in conjunction with GP IIb/IIIa receptor blockade. The mean anti-Xa level in patients undergoing PCI was 0.74 0.48 U/ml. The majority of patients who underwent PCI within eight hours of their last dose of enoxaparin had therapeutic anti-Xa levels. CONCLUSION: In patients with ACS, enoxaparin in combination with GP IIb/IIIa inhibitors and an early invasive approach resulted in comparable clinical complication and bleeding rates versus historical references utilizing unfractionated heparin. PMID- 14760190 TI - Recanalization of chronic total occlusion after conventional guidewire failure: guided by optical coherent reflectometry and facilitated by radiofrequency energy ablation. AB - The success rate of percutaneous recanalization of chronic total occlusion remains low. The Safe-Cross wire, which utilizes the principle of optical coherent reflectometry for guidance and has the ability to deliver radiofrequency energy for tissue ablation, was evaluated in 21 patients. This wire was used only after conventional guidewire failure, except in 4 patients who had failed an interventional procedure 3 months to 2 years previously. Conventional guidewires were successful in 9, and the Safe-Cross wire was successful in 10 of the remaining 12, including those 4 with a failed previous attempt. The total success rate was 90% (19/21 patients). Technological improvement in the steerability of the Safe-Cross wire may help to improve the success rate further. PMID- 14760191 TI - Treatment of chronic total occlusions using optical coherent reflectometry and radiofrequency ablative energy: incremental success over conventional techniques. PMID- 14760192 TI - Long-term outcome of percutaneous coronary interventions following failed beta brachytherapy. AB - BACKGROUND: Recurrent restenosis following vascular brachytherapy (VBT) has been reported in up to one-third of the patients enrolled in clinical trials. The long term outcome of repeat percutaneous intervention (PCI) after failed beta brachytherapy is currently unknown. METHODS: We retrospectively analyzed 97 consecutive patients undergoing percutaneous coronary reintervention after failed beta-brachytherapy at our institution (80.8% of all brachytherapy failures). Long term incidence of major adverse cardiac events (MACE, death, myocardial infarction, target lesion revascularization) was assessed. RESULTS: The procedure was successful in 90 patients (92.8%). A new stent was implanted in 72% of the procedures (sirolimus-eluting stent in 16.5%). After 3 years, survival was 94.3%, survival-free from myocardial infarction was 86.7% and MACE-free survival was 66.1%. No difference was observed in MACE-free survival between patients originally treated with brachytherapy for recurrent in-stent restenosis and patients receiving irradiation for de novo lesions (68.2% de novo group versus 61.2% ISR group; p=0.6 by log rank test). Overall, a second target lesion revascularization was performed in 27 patients (27.8%) after an average of 11.2 11.2 months; 21 patients (21.6%) had restenosis, and 6 (6.2%) developed late total vessel occlusion (related to acute myocardial infarction in 2 cases). CONCLUSION: Repeat PCI is the most common choice after failed brachytherapy. This strategy appears to be a reasonable therapeutic option for this complex iterative pathology. PMID- 14760193 TI - Initial characterization of Ikari Guide catheter for transradial coronary intervention. AB - Ikari is a new guide catheter for transradial intervention (TRI) that produces stronger back-up force by utilizing an unfavorable angle between the subclavian and brachiocephalic arteries. We report the initial results of the Ikari guide catheter based on the experience of a single center. Six operators performed a total of 102 coronary interventions for 91 patients using the Ikari guide catheter, while 101 interventions were performed with the transfemoral approach (TFI) during the same period. A left Ikari catheter was used in 63 procedures, and a right Ikari catheter was used in 39. The success rate for the procedure was 97% with a 6 French Ikari catheter. All failures were due to tortuous brachiocephalic arteries. For the Ikari procedure, the average fluorescence time was 14.5 9.5 minutes and the dye volume used was 153 53 ml; these results were equal to or better than those of TFI during the same period (20.1 12.2 minutes and 184 61 ml, respectively). These preliminary data suggest that an acceptable success rate can be achieved in TRI using appropriate guides, such as an Ikari catheter. PMID- 14760194 TI - Improved procedural results in coronary thrombosis are obtained with delayed percutaneous coronary interventions. AB - We investigated the efficacy of percutaneous coronary intervention (PCI) in coronary thrombotic lesions according to the timing of the procedure. Eighty-two patients who underwent immediate PCI (IPCI) were compared to 24 patients who underwent PCI 4.9 3 days after the diagnostic catheterization [delayed PCI (DPCI)]. DPCI was associated with a lower rate of thrombus-related angiographic events (4% versus 27%; p < 0.03), including coronary embolism (0% versus 7%; p = NS), no-reflow phenomenon (0% versus 8%; p = NS), acute closure (0% versus 10%; p = NS), stent thrombosis (4% versus 1%; p = NS) and residual thrombus (0% versus 17%; p = 0.03). No differences were seen in the hospital clinical outcome, including non-fatal myocardial infarction (4% versus 9%; p = NS), death (4% versus 0%; p = NS) or major bleeding (4% versus 3%). Delayed PCI after pharmacological treatment is a safe and efficient strategy of treatment for coronary thrombus. PMID- 14760195 TI - Intracoronary coagulative nightmare during recanalization of a recent total occlusion of the left anterior descending artery. PMID- 14760196 TI - Protruding umbrella into the left atrium associated with recurrence of a stroke: a possible late complication of the CardioSEAL occluder. AB - Percutaneous closure of a patent foramen ovale or atrial septal defect using various closure systems is now considered an alternative to surgical therapy in selected patients with cryptogenic strokes. We report the case of a late protrusion of the umbrella of a CardioSEAL Occluder into the left atrium associated with occurrence of a stroke. No recurrence of symptoms was seen at six month follow-up after reinstitution of clopidogrel in addition to aspirin. PMID- 14760197 TI - Intravascular ultrasound imaging in the diagnosis and treatment of spontaneous coronary dissection with drug-eluting stents. AB - Spontaneous coronary artery dissection is a rare cause of coronary ischemia. Its prognosis is uncertain and the optimal treatment is not fully defined. We report a case of spontaneous dissection of the left anterior descending artery in a healthy post-menopausal woman, in whom the initial strategy of medical therapy was unsuccessful. After repeated angiography, we implanted 2 drug-eluting stents using intravascular ultrasound guidance with excellent immediate result and long term symptomatic relief. This is the first report of implantation of drug-eluting stents in spontaneous coronary dissection. PMID- 14760198 TI - Treatment of long, diffuse in-stent restenosis with sirolimus-eluting stents. AB - Treatment of long, diffuse in-stent restenosis remains a therapeutic challenge. We report the successful use of multiple sirolimus-eluting stents (three 33-mm long stents) in a long, diffuse in-stent restenotic lesion. No major adverse cardiovascular events, including acute/subacute/delayed stent thrombosis, were noted at ten months. Angiography at six months showed widely patent stents with 19% restenosis only at the worst segment. PMID- 14760199 TI - Cutting balloon for bifurcation lesions jailed by NIR stent. PMID- 14760200 TI - Severe left main coronary artery spasm? PMID- 14760201 TI - Acute myocardial infarction due to left main compression aortic dissection treated by direct stenting. AB - We describe a case of acute myocardial infarction (AMI) due to compression of the left main coronary artery (LMCA) by a false channel created by an acute aortic dissection (AAD). The dynamic pattern of artery obstruction is detailed as a key element to the diagnosis of extrinsic coronary compression throughout the angiography. Treatment by direct stenting restored complete anterograde coronary flow and improved myocardial perfusion. PMID- 14760202 TI - Conservative management of ascending aortic dissection caused by percutaneous coronary intervention. AB - Despite notable refinements in percutaneous coronary intervention (PCI) techniques, these procedures are still associated with some morbidity and mortality. A case of elective coronary angioplasty is described that was complicated by coronary dissection and ST-elevation myocardial infarction. Subsequent efforts to recanalize the coronary artery resulted in a catheter induced tear at the origin of the vessel, with proximal extension of the dissection to involve the ascending aorta. Literature relating to this complication (reviewed here) suggests that surgical management may be indicated under the circumstances we describe. However, a conservative approach was adopted in this instance with excellent long-term results. PMID- 14760203 TI - Treatment of in-stent restenosis for saphenous vein grafts using intravascular brachytherapy: regulatory challenges and clinical application. AB - Treatment of in-stent restenosis using intravascular brachytherapy (IVBT) has been demonstrated to be successful and has become the standard of care for native coronary artery disease. Based on the current Food and Drug Administration (FDA) indications for use and the clinical demand to increase the scope of this form of therapy to include saphenous vein grafts for the Beta-Cath System (Novoste Corporation, Norcross, Georgia), we set out to obtain institutional approval for off-label use. Identification of institutional regulatory bodies and related procedures for obtaining off-label device use was performed. Additionally, the IVBT written directive proscription and patient informed consent forms were revised to accurately administer radiation dose and to disclose the regulatory status of using IVBT for this anatomic site. While the specifics are outlined in this report, this process and the resources needed to obtain institutional approval for off-label use are indicative of that to be expected at similar institutions. PMID- 14760204 TI - Utility of non-contact three-dimensional mapping of the left atrium for ablation of left atrial tachycardia. AB - A 78-year-old man with a highly symptomatic left atrial tachycardia, refractory to medical therapy, was referred for radiofrequency catheter ablation. Using a double trans-septal technique, two long sheaths were placed across the interatrial septum into the left atrium. Using a 64-electrode non-contact three dimensional mapping technique, the left atrium was reconstructed and the focus localized to the right superior pulmonary vein ostium. Radiofrequency energy was applied and eliminated the ectopic focus. In summary, a three-dimensional non contact mapping catheter can facilitate ectopic left atrial tachycardia ablation. PMID- 14760205 TI - How platelets work: platelet function and dysfunction. AB - This article briefly reviews (a) how platelets normally function and (b) the clinical approach to disorders of platelet numbers and function. PMID- 14760206 TI - Platelet inhibitor therapy: mechanisms of action and clinical use. AB - This article briefly reviews the mechanisms of action and clinical uses of platelet inhibitor therapy. PMID- 14760207 TI - Remodeling the blood coagulation cascade. AB - The concept of a coagulation cascade describes the biochemical interactions of the coagulation factors, but has flaws as a model of the hemostatic process in vivo. For example, the model cannot explain why hemophiliacs bleed when they have an intact factor VIIa/tissue factor ("extrinsic") pathway. Hemostasis requires the formation of an impermeable platelet and fibrin plug at the site of vessel injury, but it also requires that the powerful procoagulant substances activated in this process remain localized to the site of injury. This control of blood coagulation is accomplished by localizing the procoagulant reactions to events on specific cell surfaces to keep coagulation from spreading throughout the vascular system. A consideration of the critical role of cells allows us to construct a model of coagulation that better explains bleeding and thrombosis in vivo. This cell-based model suggests that the "intrinsic" and "extrinsic" pathways are in fact not redundant systems, but operate in parallel on different cell surfaces. PMID- 14760209 TI - Physicians' attitudes and the use of oral anticoagulants: surveying the present and envisioning future. AB - Atrial fibrillation has the highest prevalence in the elderly. While the elderly are at the highest risk for stroke and would benefit the most from anticoagulation, they are also the least likely to receive anticoagulation. In a pooled analysis of the primary prevention trials, warfarin reduced stroke by 68% compared with placebo, and aspirin reduced stroke by 18%. Age, history of hypertension, diabetes, heart failure or reduced left ventricular function, and previous transient ischemic events and stroke are independent risk factors for stroke in patients with atrial fibrillation. Less than 50% of the elderly who have clear-cut indications and no contraindications for warfarin receive anticoagulant therapy. This low use of warfarin is driven by many factors, but physicians' fear of hemorrhage is among the most important. Better adherence to evidence-driven guidelines, better patient and physician education, point-of-care monitoring of INR and the future development of user friendly anticoagulant drugs are likely to result in higher rates of anticoagulation use. PMID- 14760208 TI - Cancer and thrombosis: mechanisms and treatment. AB - Standard venous thromboembolic event (VTE) treatment practices including the use of intravenous unfractionated heparin (UFH) for initial anticoagulation, oral warfarin for chronic anticoagulation, and the prescription of only 3 to 6 months total therapy may not be optimal in the setting of active cancer and ongoing anti cancer therapy. Challenges of VTE management in cancer patients include heparin resistance due to excess circulating acute-phase proteins, increased recurrence rates during and following standard-intensity warfarin therapy, limited venous access to support therapeutic monitoring, and anticoagulation intensity independent increased bleeding rates during anticoagulation. Bleeding during anticoagulation is of particular concern in the treatment of cancer patients with disease- or chemotherapy-related thrombocytopenia, central nervous system involvement, or recent invasive procedures. Low-molecular weight heparins (LMWH) have been shown to be at least as effective and safe for initial anticoagulation compared with UFH in persons with acute VTE and have gained popularity in the setting of VTE in cancer. LMWHs have the advantage of less non-specific protein binding, subcutaneous weight-based dosing without the need for monitoring in most cases, and probably less heparin-induced thrombocytopenia. Recent trials have demonstrated efficacy superiority of select LMWHs in place of oral warfarin for long-term anticoagulation in the cancer patient. The potential for anti-tumor effects and a survival advantage associated with select classes of anticoagulant agents is actively being investigated. PMID- 14760210 TI - Home monitoring of anticoagulation. AB - Small portable devices that generate a prothrombin time/INR from fingerstick capillary blood simplify warfarin management by allowing selected patients to monitor and manage their own warfarin dose. Early studies established that patients can self-test at home, with results as accurate as those obtained by practitioners. Point-of-care testing of elderly patients resulted in tighter INR control and a lower incidence of major hemorrhage, especially at the initiation of anticoagulant therapy. Patients can also successfully self-manage warfarin therapy. Larger, prospective, randomized intervention studies have shown that patient self-management led to greater time spent within the therapeutic INR range. However, a shift toward patient self-testing will likely require centralized implementation of patient education, training, and follow up that will need to be established in the clinic setting or by a third party. PMID- 14760212 TI - Preparing for the new anticoagulants. AB - New anticoagulant agents with differing properties, risks, and potential economic consequences are becoming available. Two subcutaneously injected synthetic pentasaccharide anticoagulants, fondaparinux and idraparinux, employ the minimal chain length required to bind to antithrombin and confer a conformational change, increasing its ability to catalyze inactivation of activated factor X (Xa). Melagatran has also been developed as an injectable drug for markets outside the United States. Recently synthesized oral anticoagulants other than VKAs have proven effective in clinical trials. These drugs do not act by enhancing antithrombin activity, but rather act directly to inhibit the active site of thrombin (melagatran and its orally absorbed derivative, ximelagatran) or factor Xa (razaxaban [DPC-906]). Several studies have been performed with ximelagatran, an oral synthesized pro-drug developed by AstraZeneca, which is rapidly metabolized into the active agent, melagatran. PMID- 14760211 TI - Sepsis and disseminated intravascular coagulation. AB - Sepsis almost invariably leads to hemostatic abnormalities, ranging from insignificant laboratory changes to severe disseminated intravascular coagulation (DIC). There is compelling evidence from clinical and experimental studies that DIC is involved in the pathogenesis of microvascular dysfunction and contributes to organ failure. In addition, the massive and ongoing activation of coagulation, may deplete platelets and coagulation factors, which may in turn cause bleeding. Recent insights into important pathogenetic mechanisms that may lead to DIC have resulted in novel preventive and therapeutic approaches to patients with sepsis and a derangement of coagulation. Thrombin generation proceeds via the (extrinsic) tissue factor/factor VIIa route and simultaneously occurring depression of inhibitory mechanisms, such as antithrombin III and the protein C system. Also, impaired fibrin degradation, due to high circulating levels of PAI 1, contributes to enhanced intravascular fibrin deposition. Supportive strategies aimed at the inhibition of coagulation activation may be justified on theoretical grounds and have been found to be beneficial in experimental and initial clinical studies. These strategies comprise inhibition of tissue factor-mediated activation of coagulation or restoration of physiological anticoagulant pathways, by means of the administration of antithrombin concentrate or recombinant human activated protein C. PMID- 14760213 TI - The future of anticoagulation clinics. AB - Anticoagulation therapy is the foundation of treatment for thromboembolic disorders; and coumarin derivatives (warfarin in the United States) are the only orally administered anticoagulant medications currently available. Due to the expense and relative difficulties associated with this route of administration, parenteral drugs are not used routinely for long-term therapy, leaving warfarin as the anticoagulant of choice in the outpatient setting. The management of warfarin is problematic, however, due the nuances of its pharmacodynamic and pharmacokinetic profile and the requirement for frequent monitoring of blood levels. Although management by anticoagulation clinics is considered the gold standard for warfarin therapy, management by an anticoagulation clinic may not be the optimal option from a clinician's view and, in many cases, may not be an option at all. Anticoagulation clinics may impinge on the doctor-patient relationship. Difficulties of communication and reimbursement are not ameliorated by a specialty clinic. Innovations in warfarin management, including patient self management and computerized dosing programs, are alternatives for improved care that are available with or without input by an anticoagulation service. New oral drugs on the horizon do not require the same intensity of monitoring and do not present the same pharmacodynamic problems associated with warfarin. Warfarin will become obsolete in the foreseeable future. If anticoagulation clinics continue, they must re-define their role as the major part of the workload, warfarin management, disappears. To adapt, clinics must strengthen and enhance their role as coordinators and educators, and less so, managers of anticoagulation therapy. PMID- 14760214 TI - The future of anticoagulation clinics. AB - The emergence of novel antithrombotic agents may have a dramatic impact on anticoagulation clinics and other providers of anticoagulation services. Of the novel agents currently in development, the oral direct thrombin inhibitor ximelagatran shows the most promise as an agent to improve the field of oral anticoagulation management because it is easier and more convenient to use than warfarin. Ximelagatran is currently being investigated for several indications, including prophylaxis of venous thromboembolism in patients undergoing orthopedic surgery, venous thrombosis treatment, stroke prevention in atrial fibrillation, and acute coronary syndromes. Anticoagulation clinics presently provide systematic, organized management of oral anticoagulation therapy (warfarin in the US) and provide better patient outcomes than usual medical care. The introduction of ximelagatran in patient management may dramatically alter the workload dynamics in anticoagulation clinics. Clinics that will survive the introduction of novel agents will most likely shift from a main focus of warfarin monitoring to thrombotic disease management and coordination of all antithrombotic therapy. Comprehensive Antithrombosis Centers (CAC) will most likely manage patients with thrombotic disorders treated with a range of anticoagulant therapies. PMID- 14760215 TI - An impatient patient's perspective. PMID- 14760216 TI - Vitamin K for the treatment of asymptomatic coagulopathy associated with oral anticoagulant therapy. AB - Patients with asymptomatic elevated International Normalized Ratios (INRs) are commonly seen in practice, but there is no consensus on how best to manage this condition. Evidence suggests that low-dose (1 mg to 2.5 mg) oral vitamin K restores patients to INR values associated with a lower risk of hemorrhage more rapidly than discontinuing warfarin alone. Vitamin K therapy remains under utilized despite evidence for its effectiveness. The studies discussed in this review suggest that vitamin K1 should be considered if rapid reductions in the INR are desired. For most rapid corrections in the INR, vitamin K should be administered by the intravenous route since it begins to reduce the INR within 8 hours. Subcutaneous vitamin K is relatively ineffective, and its use may be associated with over-correction of the INR. PMID- 14760218 TI - "Bridging" therapy with low molecular weight heparin in pregnant patients and patients with mechanical prosthetic heart valves. AB - A substantial body of published, peer-reviewed, trial and cohort study-based evidence, institutional data sets and expert clinical experience/opinion supports the safe and effective use of enoxaparin for anticoagulation management of non pregnant patients with prosthetic mechanical heart valves. A comparable body of data and trial results exists, also supported by a significant and authoritative base of expert opinion, for enoxaparin-based VTE prevention and treatment of at risk pregnant patients who do not have mechanical heart valves. In pregnant women with prosthetic mechanical heart valves, no recommendations on the use of LMWHs can be made until the availability of more data (Lovenox Injection (package insert). Available at www.aventis-us.com/PIs/lovenox. Accessed July 30, 2003. Aventis Pharmaceuticals 2003). PMID- 14760219 TI - Trends in anticoagulation management across community-based practices in the United States: the Anticoagulation Consortium to Improve Outcomes Nationally (ACTION) study. AB - Randomized trials have demonstrated the effectiveness of warfarin in preventing thromboembolism. Translating the results of randomized trials into usual care is difficult, however, as trial patients are highly selected and closely monitored. The Anticoagulation Consortium to Improve Outcomes Nationally (ACTION) study is a prospective observational assessment of anticoagulation care in 98 office-based practices and 3 hospital-based clinics in the United States. Site enrollment began in August 2000 and ended in February 2002 with 6761 patients recruited. The goals of this large prospective cohort study are (1) to gather data on complication rates associated with warfarin therapy across a broad range of patient ages and indications; (2) document trends in anticoagulation management and treatment practices; and (3) establish quality benchmarks for anticoagulation control and frequency of monitoring. This report describes the methodology and preliminary results. Analysis is ongoing, and full results will be reported in the future. PMID- 14760217 TI - Pharmacogenetics and anticoagulant therapy. AB - Warfarin and other coumarins are metabolized by the cytochrome P450 2C9 complex. Common single-nucleotide polymorphisms (SNPs) in this enzyme are associated with an exaggerated elevation in the INR during warfarin initiation and an increased risk of bleeding. These observations suggest that patients known to carry the putative SNPs should be started on lower doses of warfarin therapy or have their INR values monitored more frequently during warfarin initiation. Such clinical variables as age, body surface area, and concomitant medications also play important roles in determining the maintenance dose of warfarin. Thus, a comprehensive dosing algorithm offers the most promising approach to estimating the therapeutic dose of warfarin a priori and to preventing hemorrhage during warfarin induction. PMID- 14760221 TI - [Stumbling blocks of sports]. PMID- 14760220 TI - Hormone therapies and vascular outcomes: who is at risk? AB - Postmenopausal hormone replacement therapy (HRT), such as estrogen with a progestin (E+P), is associated with an increased risk of myocardial infarction (MI), stroke, and venous thrombosis. Subgroups of susceptible women for these clinical outcomes have not been clearly identified, although women with a prior history of venous thrombosis and women with factor V Leiden are at higher risk of venous thrombosis on HRT than others. Effects of HRT on atherosclerosis, coagulation, and inflammation have been investigated, and might improve our understanding of the pathogenesis of this drug effect. In 2 trials E+P did not alter the progression of coronary atherosclerosis, while in a third trial unopposed estradiol retarded atherosclerosis progression in the carotid arteries. HRT is associated with an increase in high-sensitivity C-reactive protein (CRP), an inflammation marker associated with arterial disease risk, and an increase in activated protein C resistance, the biochemical defect associated with factor V Leiden. Given recent data, the only current indication for E+P is the short-term treatment of symptoms of estrogen deficiency, such as hot flashes. Testing for coagulation disorders or inflammatory factors, such as factor V Leiden or CRP, for use in decision-making about HRT use would be premature in unselected patients. Further research is needed to identify pathophysiological mechanisms of vascular harm from these hormones. PMID- 14760222 TI - [Effect of maintenance treatments on social life. A study with general practitioners]. AB - The objective of this retrospective study was to describe drug maintenance treatments (DMT) in ambulatory care, and to evaluate their impact on social life. A convenience sample of 665 patients on DMT was recruited by 94 general practitioners in three geographic areas (Ile-de-France, Alsace, Aquitaine), and interviewed face to face by independent staff with a standardized questionnaire on three periods (one month before DMT, 6 months after beginning of DMT and 1 month before inclusion within the survey). Among the 665 patients, 76% were on buprenorphine, 20% on methadone, and 4% on morphine sulfate. Consumption of heroin and other psychoactive drugs was lower 6 months after DMT initiation. Patients were more likely to have relationships with people who did not have any problem with alcohol and drugs after DMT initiation than before treatment. They were also more likely to spend their free time alone after having begun DMT than before. All markers of social vulnerability evaluated through standardized questionnaires (employment, housing, social insurance, days of in-patient treatment related to drug consumption and number of condemnations) were improved after a six-month period with DMT. This research contributes to demonstrating the positive impact of DMT both on drug consumption and social life. Health professionals should include social support in their clinical practice. PMID- 14760223 TI - [Sports practices and violent behaviors in 14-16 year-olds: analysis based on the ESPAD 99 survey data]. AB - To date, there has been little research into the relationship between violent behavior and the practice of sport in young adolescents (both girls and boys) in the general population. Indeed, sport is often recommended as a means of prevention and an alternative to violence in adolescence. For this reason, we studied this issue in a representative sample of 14-16 year-olds (ESPAD 99). Among this sample, boys practised some form of extra-curricular sport more than girls (75% vs. 57%), this practice being more intensive (13% boys vs. 4% girls practising sport more than 8 hours per week outside of school) and more "competitive" (14% vs. 4% taking part in national and/or international events). Boys also exhibited more violent behavior than girls, including stealing (mean 0.91 for boys vs. 0.49 in girls), fighting (mean 1.71 in boys vs. 0.49 in girls) or serious offences (mean 0.66 in boys vs. 0.34 in girls). Taking part in competitions was found to be highly associated with violent behavior. In particular, competition at a national or international level was associated (in decreasing order of significance) with fighting (OR=2.35), serious offences (OR=1.78) and stealing (OR=1.58), after adjustment for age, gender, father's educational level and type of school attended. The practical implications of this study are that it is important to moderate one's judgment about the positive effects of sport in reducing youth violence, to be attentive to violent acts perpetrated in stadiums and to better analyze the eventual beneficial effects of sports clubs. PMID- 14760224 TI - [French nationwide survey of abstinence-oriented treatments in opiate-addicted patients. Results at 12 months]. AB - Over the last few years, general practitioners and pharmacists in France have become more directly involved in the treatment of opiate-addicted patients with the rapid development of office-based buprenorphine and methadone maintenance programs. At the same time, demand for abstinence-oriented interventions outside established maintenance programs continues to be addressed to the primary care system. METHOD: Our prospective and multicentric survey was conducted to monitor the follow-up of such abstinence-oriented interventions during a 12 months period, by means of questionnaires investigating the psychosocial, medical and addictive status of the patients recruited. RESULTS: One hundred and sixty five general practitioners have accepted to participate in the survey. Initially, they recruited 414 patients (51% terminating a buprenorphine maintenance treatment, 5% a methadone treatment and 36% withdrawing from heroin). Naltrexone chlorhydrate was prescribed for 50% early after entry in the survey. During the 12-month follow-up period: four data collections were proposed at 1, 3, 6, and 12 months. In all, data were collected for 158 patients. No data could be collected for 63 patients who were excluded from follow-up analysis. Out of the 351 patients who attended at least one follow-up visit, 137 (one third) moved to an opiate agonist maintenance program. When naltrexone was prescribed, total duration of treatment averaged 4785 days with no significant difference in terms of heroin relapse and attendance to follow-up visits. Between the first and the last data collection, out of the 113 HCV-positive patients, 13 received/started an anti-viral treatment. No significant difference was noted according to indicators of social outcome. CONCLUSION: Our survey shows that office-base abstinence-oriented interventions in opiate-addicted patients can produce a moderate benefit. Research concerning alternative formulations for opiate antagonists (e.g., long acting naltrexone) could be helpfully in developing new options for treatment. PMID- 14760225 TI - [Development of diagnosis criteria for physical activity abuse and dependence: a qualitative study]. AB - AIM: The purpose of this work was to search for criteria of physical activity abuse and dependence, patterned after abuse and dependence criteria for psychoactive substances described in DSM IV and CIM10, and to determine whether these criteria could be found in individuals practising sports. This preliminary work was designed as the first step in the search for a possible physical activity dependence syndrome. METHODS: Operational criteria for physical activity abuse and dependence were defined by using the DSM IV and CIM 10 criteria for psychoactive substances abuse and dependence as a pattern. We searched for these criteria among a group of 8 persons (4 men and 4 women, aged 17-25 years) who practised 7 different sports. The subjects participated in a qualitative interview during which the defined criteria were not mentioned directly. The script of the interview was analyzed with a pre-established grid in order to search for the operational diagnostic criteria. RESULTS: All of the defined operational criteria except two concerning physical activity abuse were identified in the interview scripts. All of the participants fulfilled at least one of the defined criteria. The importance of physical activity dependence criteria spontaneously mentioned by the participants was notable; abuse criteria were found less often. DISCUSSION: Based on the findings of this qualitative study, the development of diagnostic criteria for physical activity abuse or dependence patterned after psychoactive abuse and dependence criteria appears to be a realistic possibility. The operational criteria we defined were found in all of the participants in this study, irrespective of the sports they practised. Although a large number of dependence criteria were found for each participant, this study was not designed to determine the number of participants for whom the diagnosis of physical activity dependence could be retained. This would require search for notions such as duration of the manifestations, changing functional behavior, or clinically significant suffering in order to identify the pathological nature of the behavior. Other studies are now needed to determine whether individual subjects who present a sufficient number of criteria simultaneously can be identified as presenting physical activity abuse or dependence. PMID- 14760226 TI - [Doping: health risks and relation to addictive behaviors]. AB - The paper presents the health hazards of the major doping substances and raises some questions about the relationship between doping and addictive behavior. AIMS: Current definitions of doping and addictive behavior are examined. The paper's goal is: 1- to assess the risks of neurotoxicity and overall toxicity of doping substances: stimulants, narcotics (seldom used as doping substances), and hormones, and assess their addictive potential; 2- to present available data on drug-dependent patients with a record of early prolonged and intensive physical activity or athletic practice. RESULTS AND DISCUSSION: Some doping substances present high risks for health at large doses, but usually low addictive potential and neurotoxicity. Dependency on doping substances and drift towards dependency to addictive drugs, if any, are therefore determined by genetic and environmental factors. A significant susceptibility to drug dependence has been observed in some cases of very intensive and competitive practice. Over-representation of intensive and competitive athletic antecedents among some drug-dependent patients could be accounted for in either of two ways. On the first account, the causal factor is a sensation-seeking character trait, with a likely genetic component, which predisposes the individual to the use of drugs or doping substances, as the opportunities arise. On the second account, the sudden interruption of intensive practice, and of the associated organic stress and hypersensitization of the hedonic pathway, creates a weaning syndrome and leads to the search for relief through drugs. Further exploration of this hypothesis is called for. PMID- 14760227 TI - [Misuse of tianeptine: five cases of abuse]. AB - Five cases of excessive consumption of tianeptine suggest possible drug-abuse of this substance. This side effect is unknown in animals and humans. According to DSM IV, CIM 10 criteria and the French public health code, these five patients had pathological profiles of psychoactive drug abusers. Tianeptine dosage was always used higher than recommended and the drug was taken in association with other psychotropes. Withdrawal was difficult and induced anxiety and other disorders which led to relapse in most of the patients. PMID- 14760228 TI - [Interactions between benzodiazepines and opioids]. AB - Benzodiazepines and opiates or opioids are used concomitantly in various circumstances, for example in anesthesiology, for the management of acute or chronic pain and for substitution therapy in heroin addiction. There are numerous interactions between these two families of substances. The objective of this review is to present the interactions identified in clinical and experimental studies reported in the literature dealing with their effects on pain, anxiety, sedation and respiration. The exact mechanism of benzodiazepine and opioid interactions remains to be established. It may depend on pharmacokinetic or pharmacodynamic mechanisms. Certain arguments would support the pharmacodynamic hypothesis: the co-location of GABA and opiate receptors in the central nervous system, the existence of possible cross-reactivity and common pathways of intracellular transduction. The deleterious interaction of benzodiazepines and opioids on respiration may take place at the level of the central command of ventilation or may be related to additive actions on the different neuromuscular components of the respiration. A better understanding of the exact mechanisms implicated in these interactions would increase the safety of prescription of these drugs. PMID- 14760229 TI - [Opioid tolerance: what if it was just a question of receptor internalization?]. AB - Despite side effects which appear upon chronic administration of opioid agonists, these drugs remain widely used and effective for pain relief. Among these side effects, tolerance is the major factor limiting the effectiveness of these drugs. Desensitization, defined by a decrease of opioid receptor transduction, would be a crucial step in the development of tolerance. Molecular mechanisms of opioid receptor desensitization have been extensively studied on cellular models and have been found to involve various proteins and different steps such as receptor phosphorylation, internalization, and recycling or degradation into intracellular compartments. In the present review, we discuss the role of opioid receptor internalization and sorting in desensitization and tolerance processes. PMID- 14760230 TI - [Psychological aspects of MDMA (ecstasy) users]. AB - We reviewed the literature on the psychological aspects of MDMA consumption. The present paper examined the characteristics, psychological and psychopathological consequences of synthetic drug use (MDMA). The most frequent features are depression, anxiety, cognitive disorders and sensation seeking. Longer-term, higher dosage, and use of other substances are correlated with higher risk of developing psychopathological disorders. Care should be taken in cross-sectional studies in interpreting signs and symptoms of mental disorder merely as a consequence of MDMA use, because the use of ecstasy may be associated with use of multiple substances from which the mental disorder might be more likely to precede. The consumption pattern of ecstasy and related substances must be carefully analyzed. Knowledge of the MDMA consumption profile is an important for understanding the psychological characteristics of synthetic drug users. PMID- 14760247 TI - Women in pediatric subspecialties. PMID- 14760248 TI - Early morphologic changes in the lungs of asymptomatic infants and young children with cystic fibrosis. PMID- 14760249 TI - Obesity and asthma in children. PMID- 14760250 TI - Pediatric research: what is broken and what needs to be fixed? PMID- 14760251 TI - Why we need legal standards for pediatric research. PMID- 14760252 TI - Structural airway abnormalities in infants and young children with cystic fibrosis. AB - OBJECTIVES: To determine whether the airway structure of infants and young children with cystic fibrosis (CF) differs from that of normal children by using high-resolution computed tomography (HRCT) imaging. Study design Full-inflation, controlled ventilation HRCT images of the lungs were obtained at four anatomic levels in 34 infants with CF (age, 2.4+/-1.4 years) and 20 control infants (age, 1.8+/-1.4 years). Short axis diameters of all clearly identifiable, round airway/vessel pairs were measured to obtain airway wall thickness (AWT), airway lumen diameter (ALD), and vessel diameter (VD). RESULTS: In infants with CF, mean AWT (+/-SD) was 0.58+/-0.13 mm, ALD was 1.31+/-0.56 mm, and VD was 1.62+/-0.58 mm. In control infants, mean AWT was 0.49+/-0.13 mm, ALD was 1.07+/-0.42 mm, and VD was 1.86+/-0.64 mm. Mean AWT and ALD were greater in children with CF than in normal subjects (P<.001). ALD:VD ratios increased with age in patients with CF compared with control subjects (P=.026). CONCLUSIONS: The airways of infants and young children with CF have thicker walls and are more dilated than those of normal infants. PMID- 14760253 TI - Is obesity associated with asthma in young children? AB - OBJECTIVE: The aim of this study was to evaluate the association between obesity and asthma in a population-based sample of Canadian children. STUDY DESIGN: Baseline data from the National Longitudinal Survey of Children and Youth were used in this cross-sectional study. We included 11199 children age 4 to 11 years whose biological mother reported data on asthma, height, and weight. Body mass index was categorized, and obesity was defined as body mass index >or=85th percentile. Children with asthma had parents who reported the diagnosis, and they took prescribed inhalants, had wheezing or an attack in the previous year, or had their activities limited by asthma. Multiple logistic regression was used. RESULTS: The prevalence of asthma was 9.9%. Maternal history of asthma was a risk factor for asthma among all children. Single child status and maternal depression were risk factors for girls. The odds ratio for asthma, comparing highest and lowest body mass index categories, was 1.02 (99% confidence interval, 0.70-1.46) for boys and 1.06 (99% confidence interval, 0.67-1.69) for girls. CONCLUSION: This study suggests that there is no statistical association between obesity and asthma among Canadian children age 4 to 11 years. PMID- 14760255 TI - Cardiac autonomic activity in methylmercury neurotoxicity: 14-year follow-up of a Faroese birth cohort. AB - OBJECTIVE: To determine whether heart function in childhood is affected by exposure to methylmercury (MeHg) from seafood. STUDY DESIGN: Prospective study of a Faroese birth cohort (N=1022). Examinations at ages 7 and 14 years included blood pressure, heart rate variability (HRV) and its frequency components of autonomic origin, and brainstem auditory evoked potentials (BAEPs). Mercury concentrations were determined in cord blood and in the child's hair. RESULTS: Both low-frequency (LF) and high-frequency (HF) activities decreased by about 25% from 7 to 14 years; they correlated well with the blood pressures. A doubling of prenatal MeHg exposure was associated with a decrease in LF and HF powers of about 6.7% (P=.04) and in the coefficient of variation of the electrocardiographic R-R interval of 2.7% (P=.04) at age 14 years. No discernible effect on blood pressure was apparent. Decreased LF variability was associated with increased latency of BAEP peak III, but adjustment for MeHg exposure substantially attenuated this correlation. CONCLUSIONS: Methylmercury exposure was associated with decreased sympathetic (LF) and parasympathetic (HF) modulation of the HRV. Parallel MeHg-related delays of BAEP latencies may be caused by underlying MeHg neurotoxicity to brainstem nuclei. PMID- 14760257 TI - Delayed brainstem auditory evoked potential latencies in 14-year-old children exposed to methylmercury. AB - OBJECTIVE: To determine possible exposure-associated delays in auditory brainstem evoked potential latencies as an objective measure of neurobehavioral toxicity in 14-year-old children with developmental exposure to methylmercury (MeHg) from seafood. STUDY DESIGN: Prospective study of a birth cohort in the Faroe Islands, where 878 of eligible children (87%) were examined at age 14 years. Latencies of brainstem evoked potential peaks I, III, and V at 20 and 40 Hz constituted the outcome variables. Mercury concentrations were determined in cord blood and maternal hair, and in the child's hair at ages 7 and 14. RESULTS: Latencies of peaks III and V increased by about 0.012 ms when the cord blood mercury concentration doubled. As seen at age 7 years, this effect appeared mainly within the I-III interpeak interval. Despite lower postnatal exposures, the child's hair mercury level at age 14 years was associated with prolonged III-V interpeak latencies. All benchmark dose results were similar to those obtained for dose response relationships at age 7 years. CONCLUSIONS: The persistence of prolonged I-III interpeak intervals indicates that some neurotoxic effects from intrauterine MeHg exposure are irreversible. A change in vulnerability to MeHg toxicity is suggested by the apparent sensitivity of the peak III-V component to recent MeHg exposure. PMID- 14760258 TI - Safety, efficacy, and immunogenicity of a live, quadrivalent human-bovine reassortant rotavirus vaccine in healthy infants. AB - OBJECTIVES: To investigate safety, efficacy, and immunogenicity of live quadrivalent rotavirus vaccine (QRV) containing human-bovine (WC3) reassortant rotavirus serotypes G1, G2, G3, and P1a. STUDY DESIGN: This was a randomized, double-blinded, placebo-controlled trial. During 1993 to 1994, at 10 US study sites, 439 healthy infants approximately 2 to 6 months of age, were enrolled to receive 3 doses of oral QRV or placebo at approximately 8-week intervals. RESULTS: The vaccine was generally well tolerated; no serious vaccine-related adverse experiences were reported. Risk differences and 95% confidence intervals suggested no differences between vaccine and placebo recipients in the incidences of fever, irritability, vomiting, or diarrhea during the 14 days after any dose. QRV was 74.6% efficacious (95% CI: 49.5%, 88.3%) in preventing rotavirus acute gastroenteritis (AGE), regardless of severity and 100% efficacious (95% CI: 43.5%, 100%) in preventing severe rotavirus AGE through one rotavirus season. Serotype G1 was identified in most infants with rotavirus AGE. A >or=3-fold rise in serum neutralizing antibody to G1 was observed in 57% (45/79) of vaccinees. A >or=3-fold rise in serum anti-rotavirus IgA and fecal anti-rotavirus IgA was observed in 88% (162/185) and 65% (104/159) of vaccinees, respectively. CONCLUSIONS: QRV was generally well tolerated, immungenic, and highly effective against rotavirus gastroenteritis. PMID- 14760260 TI - Prevalence and characteristics of children at increased risk for complications from influenza, United States, 2000. AB - OBJECTIVES: Annual influenza vaccination is recommended for children at high risk of complications from influenza due to underlying medical conditions, but few children are vaccinated. Vaccination also is encouraged for all children aged 6 to 23 months when feasible. This study describes the prevalence and characteristics of at-risk children nationwide. STUDY DESIGN: A descriptive analysis of the 2000 National Health Interview Survey (NHIS) was conducted among children in the United States aged 6 months through 17 years with identified high risk conditions, and among all children aged 6 to 23 months. RESULTS: Approximately 5.2 to 10.0 million children aged 6 months through 17 years (7.4% 14.2%) had high-risk conditions indicated for influenza vaccination. Asthma accounted for the majority of conditions. An estimated 7.7 million children would be aged 6 to 23 months during influenza season. Most young children and older children at high-risk have access to and frequently utilize healthcare services. CONCLUSIONS: Existing doctor visits are important opportunities for vaccinating children with high-risk conditions, or for those aged 6 to 23 months. Additional efforts are needed to implement and evaluate efficient strategies for annual influenza vaccination of children aged 6 to 23 months and for older children with medical indications. PMID- 14760261 TI - Evaluation of a urine antibody test for Helicobacter pylori in Japanese children. AB - OBJECTIVE: To evaluate the accuracy of a urine-based enzyme-linked immunosorbent assay (ELISA) kit for anti-Helicobacter pylori immunoglobulin G antibody (urine HpELISA) in children, we compared its sensitivity and specificity in reference to (13)C-urea-breath test (UBT) and H pylori stool antigen test (HpSA). STUDY DESIGN: Japanese children without significant upper abdominal symptoms were included (n=100; mean age, 7.0 years; range, 2 to 15). UBT, HpSA, and urine HpELISA were performed. RESULTS: Of 100 children, 36 and 64 were judged H pylori positive and H pylori-negative, respectively, by UBT and HpSA. Thirty-four of 36 positive children were positive by urine-HpELISA, and 62 out of 64 negative children were negative by urine-HpELISA. Thus, the urine-HpELISA had 94.4% sensitivity and 96.9% specificity, with accuracy of 96.0%. CONCLUSIONS: The urine HpELISA is a rapid, inexpensive, reliable, and easy-to-perform method for the diagnosis of H pylori infection in children. It may be useful not only for diagnosis but also for mass screening for epidemiological studies in pediatric population. PMID- 14760262 TI - Differences in antibiotic prescribing patterns for children younger than five years in the three major outpatient settings. AB - OBJECTIVES: To perform a comprehensive analysis of the use of antibiotics in three major sites for outpatient care: private office-based clinics, emergency departments (ED), and hospital-based clinics. STUDY DESIGN: Data from the National Ambulatory Medical Care Survey (NAMCS) and National Hospital Ambulatory Medical Care Survey (NHAMCS) were used to determine visit rates and antibiotic prescribing patterns for the three major outpatient care settings from 1994 to 2000 for children <5 years of age. RESULTS: Antibiotic prescription rates declined from 1405 to 1088 per 1000 children over the study years (P=.032) [correction]. Significant declines in antibiotic prescriptions were noted in both the office-based setting and ED: 1119 to 841 (P=.049) in the office-based setting and 237 to 198 antibiotic prescriptions per 1000 children in the ED (P=.003) [correction]. Sites of care differed markedly with white children receiving 82.5%, 14.3%, and 3.2% of antibiotics in the office-based settings, ED, and hospital-based clinics, respectively, compared with 60%, 31%, and 9% for black children (P<.001). However, total visits, visits resulting in a diagnosis of otitis media, and antibiotic prescribing rates were similar for white and black children during the latter study years. CONCLUSIONS: There has been a decline in antibiotic prescribing in children <5 years of age, which was most notable in office-based and emergency department settings. PMID- 14760263 TI - Chronic rhinosinusitis in young children differs from adults: a histopathology study. AB - OBJECTIVE: A histopathologic study of children with chronic rhinosinusitis (CRS) was undertaken to compare the sinus mucosa in pediatric and adult CRS. STUDY DESIGN: CRS has been defined as persistent or recurrent sinusitis symptoms for >or=12 weeks despite conventional medical therapy, with abnormal computed tomography of the maxillary sinuses. Maxillary mucosal biopsies were obtained from pediatric CRS subjects for inflammatory cell and morphologic studies. Archival sinus mucosal tissues from adults with CRS were used as histologic controls. Sinus lavages were performed on children with and without CRS for microbiologic studies. RESULTS: Sinus mucosal biopsies were obtained from 19 children with CRS (median age, 3.0 years; range 1.4-8.2 years). Pediatric CRS biopsies, as compared with adult CRS controls, had a higher density of submucosal lymphocytes (median 469 versus 294 cells/mm(2) per 5 high-power fields [HPF]; P=.02), lower density of submucosal eosinophils (medians 13 versus 82 cells/mm(2) per 5 HPF; P=.01), thinner and more intact epithelium (P=.01 and.07, respectively), thinner basement membranes (P=.002), and fewer submucosal mucous glands (P=.004). CONCLUSION: The sinus mucosa of young children with CRS has less eosinophilic inflammation, basement membrane thickening, and mucus gland hyperplasia characteristic of adult CRS. PMID- 14760264 TI - Childhood habit cough treated with self-hypnosis. AB - OBJECTIVES: To better understand factors associated with the development and persistence of habit cough and to report use of self-hypnosis for this condition. STUDY DESIGN: A retrospective chart review was performed for 56 children and adolescents with habit cough. Interested patients were instructed in self hypnosis for relaxation and to help ignore the cough-triggering sensation. RESULTS: The patients' mean age was 10.7 years. The cough was triggered by upper respiratory infections in 59%, asthma in 13%, exercise in 5%, and eating in 4%. Onset of the cough occurred as early as 2 years, and its average duration was 13 months (range, 2 weeks to 7 years). There was a high incidence of abdominal pain and irritable bowel syndrome in the 50% of the patients who missed more than 1 week of school because of their cough. Among the 51 patients who used hypnosis, the cough resolved during or immediately after the initial hypnosis instruction session in 78% and within 1 month in an additional 12%. CONCLUSIONS: Habit cough is triggered by various physiologic conditions, related frequently to other diagnoses, and it is associated with significant school absence. Self-hypnosis offers a safe efficient treatment. PMID- 14760265 TI - Prediction of the development of tolerance to milk in children with cow's milk hypersensitivity. AB - OBJECTIVES: To investigate whether the development of tolerance to cow's milk (CM) by aged 4 years can be predicted with a skin prick test (SPT) and measurements of total or specific immunoglobulin E (IgE) in the serum, taken at the time of diagnosis of cow's milk hypersensitivity (CMH). STUDY DESIGN: Infants with immediate (n=95) or delayed (n=67) challenge reactions to CM were prospectively followed to aged 4 years. CMH status was assessed annually by CM challenges. RESULTS: By aged 2, 3, and 4 years, children with delayed reactions developed tolerance to CM faster than those with immediate reactions: 64%, 92%, and 96% versus 31%, 53%, and 63%, respectively. A wheal size of <5 mm in SPT correctly identified 83% of 124 infants who developed tolerance to CM by aged 4 years, and a wheal size of >or=5 mm in SPT correctly identified 71% of 39 infants with persistent CMH. Milk-specific IgE <2 kU/L correctly identified 82% of infants who developed tolerance to CM, and milk-specific IgE >or=2 kU/L correctly identified 71% of infants with persistent CMH. CONCLUSION: SPT and milk-specific IgE in the serum are useful prognostic indicators of the development of tolerance to CM in infants with CMH. PMID- 14760266 TI - Oxygenation and hemodynamics in left and right cerebral hemispheres during induction of veno-arterial extracorporeal membrane oxygenation. AB - OBJECTIVE: Oxygenation and hemodynamics in the left and right cerebral hemispheres were measured during induction of veno-arterial extracorporeal membrane oxygenation (VA-ECMO). STUDY DESIGN: Using near infrared spectrophotometry, effects of right common carotid artery (RCCA) and right internal jugular vein (RIJV) ligation and start of VA-ECMO on concentrations of oxyhemoglobin, deoxyhemoglobin, and cerebral blood volume (CBV) were evaluated in 10 newborn infants. Mean cerebral blood flow velocity (CBFV) in the major cerebral arteries was compared before and after the start of VA-ECMO (pulsed Doppler ultrasonography). RESULTS: RCCA ligation caused a decrease in oxyhemoglobin concentration and an increase in deoxyhemoglobin concentration. RIJV ligation caused no changes. Sixty minutes after the start of VA-ECMO, oxyhemoglobin concentration and CBV had increased, and deoxyhemoglobin concentration had decreased. There were no differences between the hemispheres. Mean CBFV had increased in the left internal carotid artery, and it increased equally in both middle cerebral arteries. Flow direction was reversed in the right internal carotid artery. Three patients had asymmetric cerebral lesions, not related to differences in the measurements between the cerebral hemispheres. CONCLUSION: The initiation of VA-ECMO causes changes in cerebral oxygenation and hemodynamics but without a difference in effect on left and right cerebral hemispheres. PMID- 14760267 TI - The effect of participation in the WIC program on preschoolers' diets. AB - OBJECTIVE: To evaluate nutrient, food intake, and snacking behavior by participation in the WIC (Special Supplemental Nutrition Program for Women, Infants, and Children) program. STUDY DESIGN: Secondary data analysis of a nationally representative cross-sectional survey conducted by USDA in 1994 to 1996 and 1998. METHODS: Statistical analysis was performed correcting for sample design effects and weighting for children in two income groups (<130%, n=1772 and 130% to 185% of poverty, n=689). RESULTS: Among WIC participants, the prevalence of snacking was significantly lower (68%) compared with nonparticipants (72%) (chi(2)=5.9, P=.01). For those <130% of poverty, WIC had a beneficial effect on the intake of fat, carbohydrates, added sugar, and fruit from the total diet as well as on added sugar from snacks. These were independent of food stamp participation. For those with higher incomes, the beneficial effects were limited to added sugar, iron density, and fruit intake for the total diet. A similar significant effect of decreased added sugar intake from snacks was also seen. CONCLUSIONS: Our results are in line with previous research showing beneficial effects of WIC participation among preschoolers, primarily for nutrients targeted by the program. This study shows that the effect can reach beyond those targeted nutrients. PMID- 14760268 TI - Effect of high maternal blood phenylalanine on offspring congenital anomalies and developmental outcome at ages 4 and 6 years: the importance of strict dietary control preconception and throughout pregnancy. AB - OBJECTIVES: The Maternal Phenylketonuria Study was designed to determine the effect of a phenylalanine (Phe)-restricted diet in reducing the morbidity on the fetus. Congenital abnormalities were noted, with the focus on the effect of congenital heart defects (CHDs) and microcephaly (MICRO) on developmental outcome at 4 and 6 years of age. STUDY DESIGN: Women with blood Phe levels >240 micromol/L (n=526; to convert micromol/L to mg/dL, divide by 60) were enrolled; 382 contributed 572 pregnancies. The women had 413 offspring examined at birth and annually. At 4 years, the McCarthy General Cognitive Index was administered, and at 6 years, the Wechsler Intelligence Scale for Children Revised was administered. RESULTS: Microcephaly was noted in 137 (33%) of the offspring, and 32 (7.7%) had CHD. Maternal blood Phe levels were higher for infants with CHD and MICRO than for infants with CHD only (P=.02). Mean Phe levels at 4 to 8 weeks gestation predicted CHD (P<.0001). The McCarthy General Cognitive Index score was lower with CHD (P=.005) and MICRO (P=.0017), as was the Wechsler Intelligence Scale for Children Revised full-scale IQ score (P=.0002 for CHD and P=.0001 for MICRO). None of the subjects who had offspring with CHD had Phe control between 120 and 360 micromol/L during the first 8 to 10 weeks of gestation. CONCLUSIONS: Women with phenylketonuria need to be educated regarding diet for life. This should help improve diet control before conception and throughout pregnancy. PMID- 14760269 TI - Maternal age and other predictors of newborn blood pressure. AB - OBJECTIVE: To investigate perinatal predictors of newborn blood pressure. STUDY DESIGN: Among 1059 mothers and their newborn infants participating in Project Viva, a US cohort study of pregnant women and their offspring, we obtained five systolic blood pressure readings on a single occasion in the first few days of life. Using multivariate linear regression models, we examined the extent to which maternal age and other pre- and perinatal factors predicted newborn blood pressure level. RESULTS: Mean (SD) maternal age was 32.0 (5.2) years, and mean (SD) newborn systolic blood pressure was 72.6 (9.0) mm Hg. A multivariate model showed that for each 5-year increase in maternal age, newborn systolic blood pressure was 0.8 mm Hg higher (95% CI, 0.2, 1.4). In addition to maternal age, independent predictors of newborn blood pressure included maternal third trimester blood pressure (0.9 mm Hg [95% CI, 0.2, 1.6] for each increment in maternal blood pressure); infant age at which we measured blood pressure (2.4 mm Hg [95% CI 1.7, 3.0] for each additional day of life); and birth weight (2.9 mm Hg [95% CI, 1.6, 4.2] per kg). CONCLUSIONS: Higher maternal age, maternal blood pressure, and birth weight were associated with higher newborn systolic blood pressure. Whereas blood pressure later in childhood predicts adult hypertension and its consequences, newborn blood pressure may represent different phenomena, such as pre- and perinatal influences on cardiac structure and function. PMID- 14760270 TI - First episode of acute CNS inflammatory demyelination in childhood: prognostic factors for multiple sclerosis and disability. AB - OBJECTIVES: To evaluate prognostic factors for second attack and for disability in children presenting with an initial episode of central nervous system (CNS) demyelination. STUDY DESIGN: A cohort of 296 children having a first episode of acute CNS inflammatory demyelination was studied by survival analysis. RESULTS: The average follow-up was 2.9+/-3 years. At the end of the follow-up, 57% of patients had a diagnosis of multiple sclerosis (MS), 29% had a monophasic acute disseminated encephalomyelitis, and 14% had a single focal episode. The rate of a second attack was (1). higher in patients with age at onset >or=10 years (hazard ratio, 1.67; 95% CI, 1.04-2.67), MS-suggestive initial MRI (1.54; 1.02-2.33), or optic nerve lesion (2.59; 1.27-5.29); and (2). lower in patients with myelitis (0.23; 0.10-0.56) or mental status change (0.59; 0.33-1.07). Of patients with a second attack, 29% had an initial diagnosis of acute disseminated encephalomyelitis. At the end of the follow-up period, 90% of patients had no or minor disability. Occurrence of severe disability was associated with a polysymptomatic onset (3.25; 1.16-11.01), sequelae after the first attack (26.65; 9.42-75.35), further relapses (1.49; 1.16-1.92), and progressive MS (3.57; 1.21 8.72). CONCLUSIONS: Risk of second attack of CNS demyelination is higher in older patients and lower in patients with mental status change. Risk of disability is higher in polysymptomatic and relapsing patients. PMID- 14760271 TI - Overdiagnosis of osteoporosis in children due to misinterpretation of dual-energy x-ray absorptiometry (DEXA). AB - OBJECTIVE: Dual-energy x-ray absorptiometry (DEXA) is increasingly used to evaluate children for osteoporosis. However, the interpretation of pediatric DEXA is complicated by growth and development. Because most DEXA scans are performed on adults, we hypothesized that physicians who interpret DEXA may not be aware of these pediatric issues, potentially leading to misdiagnosis. STUDY DESIGN: Children (n=34, aged 4-17 years) diagnosed with low bone mineral density (BMD) based on a DEXA scan were referred for possible inclusion in a childhood osteoporosis protocol. The referral DEXA scans were analyzed for accuracy. RESULTS: Thirty (88%) of the scans had at least one error in interpretation. The most frequent error (62%) was use of T-score (SD score compared with young adults) to diagnose osteoporosis, which is inappropriate for children. Other errors included use of a reference database that does not consider gender or ethnic differences (21%), incorrect bone map (21%), inattention to short stature (15%), and other measurement or statistical error (12%). After correcting for these errors, 53% had normal BMD, whereas only 26% retained the diagnosis of low BMD. The remaining 21% could not be given a definitive diagnosis. CONCLUSION: In children, the diagnosis of osteoporosis is often due to misinterpretation of a DEXA scan. PMID- 14760272 TI - A six-month-old infant with liver steatosis. PMID- 14760273 TI - Joubert-like syndrome unlinked to known candidate loci. AB - We observed the Joubert syndrome (JS) associated with bilateral morning glory disk anomaly and cystic dysplastic kidneys in three patients from a consanguineous kindred. Homozygosity mapping excluded three JS candidate loci as sites harboring the disease gene. We thus delineate an autosomal recessive disorder, distinct from JS and related conditions. PMID- 14760275 TI - Death caused by hyperglycemic hyperosmolar state at the onset of type 2 diabetes. AB - Seven obese African American youth were considered to have died from diabetic ketoacidosis (DKA) due to type 1 diabetes, despite meeting the criteria for hyperglycemic hyperosmolar state and not for DKA. All had previously unrecognized type 2 diabetes, and death may have been prevented with earlier diagnosis or treatment. PMID- 14760276 TI - IMAGe syndrome: a complex disorder affecting growth, adrenal and gonadal function, and skeletal development. AB - IMAGe syndrome (intrauterine growth restriction, metaphyseal dysplasia, adrenal hypoplasia congenita, genital abnormalities; MIM 300290) is a multisystem disorder with a broad phenotype, which, if unrecognized, may result in major and possibly life-threatening complications. Initial clinical features overlap with those of Russell-Silver syndrome (RSS) and isolated growth hormone (GH) deficiency, conditions from which it must be distinguished. We report an Australian male with adrenal hypoplasia congenita (AHC) in association with IMAGe syndrome. The patient had intrauterine growth restriction (IUGR) and dysmorphic features comprising small, low-set ears, micrognathia, bilateral cryptorchidism, micropenis, and skeletal abnormalities. Signs of adrenal insufficiency occurred at aged 4.6 years. Our patient differs from those previously described by the late onset of adrenal insufficiency and the presence of GH deficiency. IMAGe is a complex syndrome involving dysmorphic features; disorders of growth, gonadal, and adrenal function; and skeletal abnormalities. PMID- 14760277 TI - Bilateral sensorineural deafness in adenosine deaminase-deficient severe combined immunodeficiency. AB - Adenosine deaminase deficiency presents with severe combined immunodeficiency and is treatable by bone marrow transplantation. With improved survival, the nonimmunologic manifestations of this condition are becoming apparent. We report a high incidence of bilateral sensorineural deafness in transplanted patients, which highlights the systemic nature of the disease. PMID- 14760278 TI - Early onset of severe diabetes mellitus-related microvascular complications. AB - A 16-year-old girl with type 1 diabetes developed painful peripheral neuropathy within 1 month and proliferative retinopathy within 1 year despite excellent glycemic control. We speculate on potential mechanisms that may have contributed to the rapid development of diabetes mellitus-related complications. PMID- 14760280 TI - Self-mutilation and mental retardation: clues to congenital insensitivity to pain with anhidrosis. PMID- 14760281 TI - When to use bisphosphonates. PMID- 14760283 TI - Increased nocturnal heart rate in children with renal scars. PMID- 14760285 TI - Can oral midazolam provide relief of discomfort or analgesia? PMID- 14760287 TI - Noroviruses: agents in outbreaks of acute gastroenteritis. AB - Noroviruses are the most common cause of acute gastroenteritis in the United States and are believed to be the most common cause of food borne illnesses.1 Noroviruses have avoided attention for years due to the difficulty of detection and inability to be cultured. Norovirus outbreaks have major implications for health care workers as they can occur in nursing homes and hospitals. To further complicate the picture, these viruses can infect persons of all ages which is a feature that distinguishes noroviruses from other agents. Factors that contribute to the significant impact of noroviruses include a large human reservoir, low infection dose, and the ability to be transmitted by various routes. This article provides an overview of noroviruses particularly as it relates to health care workers. PMID- 14760288 TI - Earthquakes in El Salvador: a descriptive study of health concerns in a rural community and the clinical implications--part II. AB - Results reported in Part I of the Earthquakes in El Salvador series (see Disaster Management & Response 2003;1:105-9) indicated clinically relevant findings. The findings indicated a need for greater public health action within all five categories reviewed: healthcare, access to healthcare, housing, food, water and sanitation. Significant results between urban and rural communities indicated a need for broader community aid, public health and sanitation services to rural areas. Faster and more efficient disaster management and care services throughout the San Sebastian community were also necessary modifications. PMID- 14760290 TI - Providing emergency care during a power outage: August 2003. PMID- 14760289 TI - Vertical evacuation drill of an intensive care unit: design, implementation, and evaluation. AB - Hospital disaster plans should be developed to cover any potential event that could require the immediate evacuation of all patients. Intensive care unit (ICU) patients present multiple challenges for planners: reliability of hospital elevators, manpower needed to transport patients, and the time needed for stabilization before patient transfer. If the ICU is located on an upper floor of the hospital and patients have numerous life-support interventions in place, a vertical evacuation can require more resources than anticipated. A study was done using 12 moulaged patients who were carried down 4 flights of stairs. The lessons learned from the drill are reported. PMID- 14760291 TI - Joint Regional Exercise ("JREX") 2000. AB - Joint Regional Exercise, 2000 (J-REX 2000) was an ambitious, multi-functional, multi-agency exercise conducted October 12-14, 2000. While the main focus of the exercise was centered in Minneapolis MN, and adjacent Fort Snelling, the exercise also involved National Disaster Medical System (NDMS) participating hospitals in Cleveland, Detroit, Duluth, Indianapolis, Omaha, and Tulsa. As such, the exercise presented a unique opportunity to simultaneously and somewhat realistically examine the response capabilities of multiple agencies and institutions within the NDMS and to 'game' several aspects of the Federal Response Plan's Emergency Support Function 8. PMID- 14760299 TI - Airfoils for airplanes. PMID- 14760301 TI - Transporting neonates with nitric oxide: the 5-year ShandsCair experience. AB - Traditionally, hypoxic respiratory failure in the newborn has been treated with supplemental oxygen, conventional mechanical ventilation, sedation, and high frequency oscillatory ventilation. Despite appropriate management with these treatment modes, care for critically ill newborns often requires more invasive measures, including extracorporeal membrane oxygenation (ECMO). Although it may be life saving, ECMO requires cannulation and anticoagulation, which introduces significant risk of morbidity. PMID- 14760302 TI - Spirit of Marshfield: celebrating 10 years of critical care transport excellence. PMID- 14760303 TI - Operational cost and value. PMID- 14760304 TI - And 'the best job in the world award' goes to...classic lifeguard. PMID- 14760305 TI - The Red Cross Air Mercy Service: a developing air medical service in Southern Africa. PMID- 14760306 TI - Comparison of the intubating laryngeal mask airway versus laryngoscopy in the Bell 206-L3 EMS helicopter. AB - INTRODUCTION: The purpose of this study is to compare intubation success rate and time to intubation for the intubating laryngeal mask airway (I-LMA) versus direct laryngoscopy (DL) using a manikin model during a simulated in-flight scenario. SETTING: The setting for the study was a University hospital-based air medical program. METHODS: This was a prospective, randomized, crossover trial. Eight nurses and 7 paramedics were randomly assigned to perform 3 intubations with either the I-LMA or DL first and then 3 intubations using the alternate technique. Descriptive statistics, 95% confidence intervals (CIs) of means, and Fisher's Exact test were conducted for comparisons. RESULTS: Fifteen set-ups and 45 intubations were performed with each technique. Previous experience was 74 mm (95% CI 64-84) with DL and 18 mm (95% CI 9-27) with the I-LMA. Set-up time was 33 seconds (95% CI 26-40) for DL and 40 seconds (95% CI 29-50) for I-LMA. Time to intubation was 12 seconds (95% CI 10-14) for DL and 39 seconds (95% CI 31-48) for I-LMA. Success rate was 100% for DL and I-LMA placement and 98% for intubation through the I-LMA. Crew rated difficulty of DL 13 mm (95% CI 6-20), placing the I LMA 23 mm (95% CI 13-32), and intubating through the I-LMA 17 mm (95% CI 10-24). CONCLUSION: Intubation success was very high for both DL and the I-LMA, despite less flight crew experience with the I-LMA. Total time to intubation was longer with the I-LMA but still less than 1 minute. The flight crews considered both techniques easy to perform. The I-LMA appears to be a useful adjunct for airway management in the Bell 206-L3 helicopter. PMID- 14760307 TI - Inverse intubation in air medical transport. AB - INTRODUCTION: The purpose of this study was to investigate the speed and accuracy of the inverse intubation procedure. TECHNIQUE: The operator crouches or kneels over the patient, straddling the torso. The laryngoscope is held in the operator's right hand in an overhand fashion, inserted gently into the patient's mouth, and pulled up and caudad. When the vocal cords are visualized, the endotracheal tube is passed with the left hand. METHODS: The procedure was taught to flight nurses and respiratory therapists (n = 21) using an intubating mannequin strapped to a stretcher in a BK 117 helicopter to simulate an in-flight intubation. The subjects were timed intubating the mannequin using both inverse and standard intubation techniques. Each technique was performed twice, and the times were averaged. The Wilcoxon Signed Ranks test was used to determine statistical significance. RESULTS: No significant difference occurred between times of the standard (24.0 s) and inverse techniques (21.6 s) (P =.715) or number of attempts for successful endotracheal intubation (1.12 and 1.07, P =.581). CONCLUSION: Inverse intubation is a useful skill for prehospital providers. This skill can be taught in a brief period and used successfully with no compromise in speed or success rate. PMID- 14760308 TI - Cardiac surgery in octogenarians: have we gone too far...or not far enough? PMID- 14760309 TI - Training trials in heart failure: time to exercise restraint? PMID- 14760310 TI - Heart failure treatment and renal function. PMID- 14760311 TI - Left atrial size: renewed interest in an old echocardiographic measurement. PMID- 14760312 TI - Restenosis: are the implications changing? PMID- 14760313 TI - Clinical trials in Japan and the United States. PMID- 14760314 TI - Antibiotics for secondary prevention of coronary artery disease: an ACES hypothesis but we need to PROVE IT. PMID- 14760315 TI - Long-term care of the patient with the atrial defibrillator. AB - With an aging population, atrial fibrillation is becoming an increasingly common cause of hospital admission. Patients with recurrent, symptomatic persistent atrial fibrillation often require repeated admissions to the hospital for cardioversion. The development of the atrial defibrillator has empowered such patients to take charge of their condition and perform cardioversion on themselves at home. This liberates them from the worry of hospitalization and can increase patient confidence. The implantation of an atrial defibrillator, however, has some disadvantages, and long-term use of the device exposes patients to some of the psychological adaptations that occur in recipients of implantable devices. This article discusses in depth the patient selection process, the implantation procedure, the use of the atrial defibrillator, and problems that can arise during long-term follow-up. PMID- 14760316 TI - Interstudy reproducibility of right ventricular volumes, function, and mass with cardiovascular magnetic resonance. AB - BACKGROUND: Cardiovascular magnetic resonance (CMR) has shown excellent results for interstudy reproducibility in the assessment of left ventricular (LV) parameters. However, interstudy reproducibility data for the more complex-shaped right ventricle in a large study group have not yet been reported. We sought to determine the interstudy reproducibility of measurements of right ventricular (RV) volumes, function, and mass with CMR and compare it with correspondent LV values. METHODS: Sixty subjects (47 men; 20 healthy volunteers, 20 patients with heart failure, 20 patients with ventricular hypertrophy) underwent 2 CMR studies for assessment of RV measurements with a minimum time interval between each study. RESULTS: The overall interstudy reproducibility (range between groups) for the RV was 6.2% (4.2%-7.8%) for end-diastolic volume, 14.1% (8.1%-18.1%) for end systolic volume, 8.3% (4.3%-10.4%) for ejection fraction (EF), and 8.7% (7.8% 9.4%) for RV mass. RV reproducibility was not as good as for the LV for all measures in all 3 groups, but this was only statistically significant for EF (P <.01). CONCLUSIONS: CMR showed good interstudy reproducibility for RV function parameters in healthy subjects, patients with heart failure, and patients with hypertrophy, which suggests that CMR is reliable for serial RV assessment. These data can be used to power sample sizes for longitudinal research studies of RV volume and function. The reproducibility values were similar to, but generally lower than, the reproducibility values for the LV in the same study population, which indicates that sample sizes for RV studies are in general larger than those for LV studies. PMID- 14760317 TI - Prevalence of structural cardiac abnormalities in patients with myotonic dystrophy type I. AB - BACKGROUND: Myotonic dystrophy type 1 (DM1) is a neurological disorder with known cardiac involvement, including conduction disturbances, arrhythmias, and ventricular dysfunction. We studied which clinical and electrocardiographic features are associated with structural cardiac abnormalities. METHODS: History, physical examination, electrocardiography, and genetic testing were performed on 382 patients with DM1, and cardiac imaging was performed on 100 of these patients. RESULTS: Clinical congestive heart failure was found in 7 of the 382 patients (1.8%). Structural cardiac abnormalities determined with cardiac imaging included left ventricular hypertrophy (19.8%), left ventricular dilatation (18.6%), left ventricular systolic dysfunction (14.0%), mitral valve prolapse (13.7%), regional wall motion abnormality (11.2%), and left atrial dilatation (6.3%). Left ventricular systolic dysfunction was associated with increasing age (relative risk [RR], 1.9 per decade; 95% CI, 1.1-3.2; P =.02), cytosine-thymine guanine (CTG) repeat length (RR, 2.8 per 500 repeats; 95% CI, 1.3-6.3; P =.01), P R >200 ms (RR, 14.7; 95% CI, 3.0-73.1; P =.001), and QRS >120 ms (RR, 5.7; 95% CI, 1.5-21.8; P =.01). P-R >200 ms was predictive of regional wall motion abnormalities. QRS >120 ms correlated with regional wall motion abnormalities and left atrial dilatation. CONCLUSIONS: Several clinical and electrocardiographic findings in patients with DM1 are significantly associated with structural heart abnormalities. These results suggest an underlying genetic and pathophysiologic correlate that may lead to cardiac disease in these patients. PMID- 14760318 TI - Conflict of interest. AB - The clinical research enterprise is increasingly scrutinized, in part because of the issue of conflict of interest. The issue is broad and its implications touch on a wide range of concerns, from the safety of patient care to the viability of a large industry. Numerous constituencies are affected by conflict of interest, and representatives of all of them convened in November 2002 for a one-and-a-half day discussion of the issues as well as possible solutions to both the perception and the actuality of such conflict. Participants included medical journal editors, news reporters, physician investigators, representatives of institutional conflict-of-interest oversight committees, representatives of the medical products industry, and Federal regulators. The resulting manuscript provides a review of the issues as well as desirable ways for each of the players to monitor themselves; each section thus contains provocative recommendations for eliminating conflict of interest to ensure that our vibrant health care system continues to foster exciting new advances to improve patient care. PMID- 14760319 TI - Which beta-blocker for heart failure? PMID- 14760320 TI - High fasting glucose levels as a predictor of worse clinical outcome in patients with coronary artery disease: results from the Bezafibrate Infarction Prevention (BIP) study. AB - BACKGROUND: A high fasting glucose level may be a marker not only for microvascular complications, but also for macrovascular complications. We evaluated the clinical significance of a high fasting glucose level (> or =110 mg/dL), detected either at baseline or during follow-up, in the Bezafibrate Infarction Prevention (BIP) study. METHODS: The BIP study was a secondary prevention prospective double-blind study comparing bezafibrate to placebo. A total of 3122 patients with documented coronary artery heart disease who were aged 45 to 74 years and had a total cholesterol level between 180 and 250 mg/dL, low-density lipoprotein cholesterol level < or =180 mg/dL, a high-density lipoprotein cholesterol level < or =45 mg/dL, a triglyceride level < or =300 mg/dL, and a fasting glucose < or =160 mg/dL were randomized to receive 400 mg of bezafibrate daily or placebo. RESULTS: The primary end point of the BIP study was fatal myocardial infarction, non-fatal myocardial infarction, or sudden death. Secondary end points included hospitalization for unstable angina, percutaneous transluminal coronary angioplasty, and coronary artery bypass grafting. At baseline, 330 patients (11%) had diabetes mellitus, and 293 patients (9%) had an impaired fasting blood glucose level (IFG). During 6.2 years of follow-up, diabetes mellitus developed in 186 patients (6%), IFG developed in 366 patients (12%), and 62% of patients remained with normal fasting glucose levels (NFG). Patients with diabetes mellitus and IFG both at baseline or developing during follow-up had a significantly higher rate of secondary end points than paients with NFG (P <.0001). Bezafibrate treatment reduced secondary end points only in patients with NFG (P =.04). CONCLUSION: Diabetes mellitus and IFG were common in the BIP study and were predictive of a worse clinical outcome that was not attenuated with bezafibrate treatment. PMID- 14760321 TI - Association of diabetes mellitus and glycemic control strategies with clinical outcomes after acute coronary syndromes. AB - BACKGROUND: Diabetes is associated with an increased risk for coronary artery disease (CAD) and its complications. The relative effect of glucose-lowering strategies of "insulin provision" versus "insulin sensitization" among patients with CAD remains unclear. METHODS: To evaluate the associations of diabetes and hypoglycemic strategies with clinical outcomes after acute coronary syndromes, we analyzed data from 15,800 patients enrolled in the SYMPHONY and 2nd SYMPHONY trials. RESULTS: Compared with nondiabetic patients, patients with diabetes (n = 3101; 19.6%) were older, more often female, more often had prior CAD, hypertension, and hyperlipidemia, and less often were current smokers. The diabetic cohort had higher 90-day unadjusted risk of the composite of death/myocardial infarction (MI)/severe recurrent ischemia (SRI), death/MI, and death alone, as well as a near doubling of 1-year mortality rates. At 1 year, diabetes was associated with significantly higher adjusted risks of death/MI/SRI (OR, 1.3 [95% confidence interval, 1.1, 1.5]) and death/MI (OR, 1.2 [1.0, 1.4]). Hypoglycemic therapy including only insulin and/or sulfonylurea (insulin providing; n = 1473) was associated with higher 90-day death/MI/SRI compared with therapy that included only biguanide and/or thiazolidinedione therapy (insulin sensitizing; n = 100) (12.0% vs 5.0%); (adjusted OR, 2.1 [1.2, 3.7]). CONCLUSIONS: Diabetic patients with acute coronary syndromes had worse clinical outcomes. Although the findings regarding the influence of glycemic-control strategies should be interpreted with caution because of the exploratory nature of the analyses and the relatively small sample size of the insulin-sensitizing group, the improved risk-adjusted outcomes associated with insulin-sensitizing therapy underscore the need to further evaluate treatment strategies for patients with diabetes and CAD. PMID- 14760322 TI - Effectiveness of primary percutaneous coronary intervention compared with that of thrombolytic therapy in elderly patients with acute myocardial infarction. AB - BACKGROUND: Few data exist from a community-based perspective on the relative effectiveness of primary percutaneous coronary intervention (PCI) as compared with thrombolytic therapy (TT) in elderly patients with ST-elevation myocardial infarction (STEMI), particularly in the current era of coronary stents and newer antithrombotic agents. METHODS: We evaluated data from patients, aged > or =70 years, with STEMI who were enrolled in the Global Registry of Acute Coronary Events study between April 1999, and September 2002. RESULTS: Of the 2975 elderly patients eligible for reperfusion therapy, 365 (12.7%) underwent primary PCI and 769 (26.7%) received TT. The median delay from hospital arrival to therapy was 105 minutes for primary PCI and 40 minutes for TT. Inhospital complications for primary PCI versus TT included mortality (13.5% vs 14.8%), reinfarction (1.1% vs 5.7%), composite of death or reinfarction (14.3% vs 18.7%), cardiogenic shock (11.3% vs 11.6%), major bleeding (8.6% vs 5.9%), and stroke (1.1% vs 2.8%). After adjustment for baseline differences and propensity score, patients receiving primary PCI showed a lower rate of reinfarction (odds ratio [OR], 0.15; 95% CI, 0.05-0.44) and mortality (OR, 0.62; 95% CI, 0.39-0.96) and the composite of reinfarction or death (OR, 0.53; 95% CI, 0.35-0.79), with no difference in other outcome measures. CONCLUSION: Our data suggest that, compared with TT, primary PCI is associated with a decrease in reinfarction and mortality, with no change in other outcome measures, in elderly patients with STEMI. These findings from an observational registry require further confirmation in future randomized clinical trial assessing the optimal reperfusion strategy in the elderly cohort with STEMI. PMID- 14760323 TI - Activated factor 12 (FXIIa) predicts recurrent coronary events after an acute myocardial infarction. AB - BACKGROUND: Activated factor XII (FXIIa) is involved in vascular injury and repair, participating in inflammation, thrombosis, and fibrinolysis. We wanted to test the hypothesis that FXIIa may predict an acute coronary syndrome (ACS) after a myocardial infarction (MI) and to evaluate whether FXIIa is related to global markers of end-stage coagulation and inflammation, including fibrin monomer (FM) and ultrasensitive C-reactive protein (microCRP). METHODS: In a prospective study of 300 patients with acute MI, we evaluated the predictive value of FXIIa in blood samples drawn 4 to 6 days after admission. Cardiac death, re-MI, and troponin-T-positive unstable angina pectoris were registered during a median follow-up period of 1.5 years. RESULTS: In the upper quartile of FXIIa (Q4) (> or =2.23 ng/mL) 32.0% of patients had an ACS as compared with 16.9% of patients with FXIIa in the three lower quartiles (Q1-3, P =.008). Relative risk of recurrent ACS for patients with FXIIa in the Q4 as compared with Q1-3 was 1.89 (95% CI, 1.22 to 2.93). A secondary ACS occurred earlier in patients with FXIIa in the Q4 as compared with those with FXIIa in the Q1-3 (P =.0039). Conventional risk factors as potential confounders were not associated with time to event. FXIIa did not correlate with FM or microCRP, and the FM and microCRP levels were of a similar magnitude in the Q4 as compared with the Q1 and the Q1-3 of FXIIa. CONCLUSIONS: FXIIa predicts recurrent coronary events after MI. The prognostic ability of FXIIa was not reflected by markers of hypercoagulability or inflammation. PMID- 14760324 TI - Activation of the inflammation, coagulation, and fibrinolysis systems, without influence of abciximab infusion in patients with non-ST-elevation acute coronary syndromes treated with dalteparin: a GUSTO IV substudy. AB - BACKGROUND: In acute coronary syndromes, the inflammation and the coagulation systems are activated, implying an impaired outcome. In addition to platelet inhibition, recent evidence suggests that the glycoprotein IIb/IIIa receptor inhibitor abciximab attenuates inflammation and coagulation activity. METHODS: The Swedish Global Utilization of Strategies To open Occluded arteries-IV (GUSTO IV) substudy included 404 patients with non-ST-elevation acute coronary syndromes. In addition to aspirin and dalteparin, all patients were randomized to receive abciximab infusion for 24 hours or 48 hours or corresponding placebo without early coronary revascularization. Plasma samples were obtained at baseline and 24, 48, and 72 hours. RESULTS: The median levels of the coagulation markers thrombin/antithrombin complex and soluble fibrin increased significantly from 3.1 to 3.7 ug/L (baseline to peak; P <.001) and from 20 to 23 nmol/L (P <.001), respectively. The fibrinolysis marker, tissue plasminogen-activator, also increased its median levels, from 11.7 to 17.5 ug/L (P <.001), whereas the median level of plasminogen-activator-inhibitor was unchanged. The inflammatory markers interleukin-6, C-reactive protein, and fibrinogen also increased their median levels (5.4-7.8 ng/L, P <.001; 4.4-8.7 mg/L, P <.001; 3.3-3.9 g/L, P <.001). However, there were no differences in median levels or in changes of median levels of any marker at any point between the placebo group and any of the abciximab groups. CONCLUSIONS: In non-ST-elevation acute coronary syndrome, there was a simultaneous activation of the inflammation, coagulation, and fibrinolysis systems, despite aspirin and dalteparin treatment. Prolonged treatment with abciximab had no influence of the activation of these systems. PMID- 14760325 TI - Single lead ST-segment recovery: a simple, reliable measure of successful fibrinolysis after acute myocardial infarction. AB - BACKGROUND: Successful reperfusion after acute ST-elevation myocardial infarction improves prognosis. Among the different electrocardiographic markers of reperfusion, sum ST resolution is considered the hallmark of reperfusion, but is cumbersome to use. METHODS: To assess the usefulness of a single lead ST resolution at 90 minutes after fibrinolysis compared with the sum ST resolution in predicting Thrombolysis in Myocardial Infarction (TIMI) grade 3 flow, we used prospectively collected data from the Limitation of Myocardial Injury Following Thrombolysis in Acute Myocardial Infarction (LIMIT-AMI) study. All patients had electrocardiograms recorded at presentation and 90 minutes and a coronary angiogram 90 minutes after fibrinolysis. RESULTS: Infarction artery patency was assessed in 238 patients with 4 different ST resolution criteria: single lead ST resolution > or =50% and > or =70% and sum ST resolution > or =50% and > or =70%. The most sensitive criteria for TIMI grade 3 flow was single lead ST resolution > or =50% (sensitivity rate, 70%; specificity rate, 54%), whereas sum ST resolution > or =70% was most the specific criteria (sensitivity rate, 45%; specificity rate, 79%). The proportion of patients with TIMI grade 3 flow was similar in all 4 ST resolution groups (P =.84). Pre-discharge infarction size and ejection fraction were also similar. No single lead or sum lead measure of ST resolution was significantly associated with an increased risk of death, heart failure, or reinfarction. CONCLUSION: We propose that single lead ST-resolution > or =50% as an optimal electrocardiographic indicator for successful reperfusion 90 minutes after fibrinolysis. This simple electrocardiographic measure should be combined with bedside clinical and hemodynamic assessment to optimize decision making after fibrinolysis. PMID- 14760326 TI - Change of multiple complex coronary plaques in patients with acute myocardial infarction: a study with coronary angiography. AB - BACKGROUND: Patients with acute myocardial infarction (AMI) may have multiple complex coronary plaques that are not limited to the culprit lesions. However, it is unknown whether they tend to progress in severity, regress, or remain stable. The aim of this angiographic study is to evaluate the natural history of these lesions. METHODS: We consecutively enrolled 229 patients who underwent coronary angiography at the time of their hospitalization to treat AMI with primary angioplasty. Baseline and follow-up (mean follow-up duration, 192 +/- 33 days) coronary angiographic data in patients with multiple complex coronary plaques characterized by thrombus, ulceration, plaque irregularity, and impaired flow were compared. RESULTS: Single complex coronary plaques were identified in 167 patients (73%), and multiple complex plaques were identified in the other 62 patients (27%). Among the patients with multiple complex plaques (62 patients, 83 non-culprit complex plaques), the angiographic examinations were reviewed simultaneously in 43.5% (27 patients, 35 non-culprit complex plaques). Of 35 non culprit complex lesions, 29 lesions (82%) remained complex without changing into smooth lesions, 1 lesion became totally occluded, and 4 lesions regressed. The severity of non-culprit complex lesions between baseline and follow-up angiography is equal (maximal diameter stenosis, 74% +/- 15% vs 72% +/- 15%, P =.4). Long-term cardiac events after discharge were more likely to develop in patients with multiple complex plaques than in patients with single complex plaques (24% vs 10%, respectively; P <.01). CONCLUSIONS: In patients with AMI, little angiographic change occurred during 6 months of follow-up in the non culprit complex plaques. PMID- 14760327 TI - Is systolic blood pressure recovery after exercise a predictor of mortality? AB - BACKGROUND: An attenuated systolic blood pressure recovery after exercise has been associated with the severity of atherosclerotic heart disease. METHODS: For 6 years, we observed 12,379 patients who underwent symptom-limited exercise testing. We excluded patients receiving antihypertensive medication and patients with valvular disease, emphysema, end-stage renal disease, heart failure, left ventricular systolic dysfunction, and atrial fibrillation. Blood pressure recovery ratio was defined as the ratio of systolic blood pressure at 3 minutes into recovery to systolic blood pressure at peak exercise; this has been shown to correlate with angiographic severity of coronary disease. RESULTS: The blood pressure recovery ratios ranged from 0.36 to 1.62, with values for increasing quartiles of 0.72 +/- 0.05, 0.82 +/- 0.02, 0.88 +/- 0.02, and 0.99 +/- 0.07. During follow-up, there were 430 deaths (3%). Five-year Kaplan Meier survival rates were 0.975, 0.974, 0.969, and 0.966 in quartiles 1 to 4, respectively. Compared with patients in the lowest quartile of blood pressure recovery ratio, patients in the highest quartile were at somewhat increased risk (hazard ratio, 1.71; 95% CI, 1.31-2.24; P <.001). However, after adjusting for age, sex, body mass index, resting heart rate and blood pressure, peak systolic blood pressure, heart rate recovery, exercise chronotropic response, cardiac history, and standard risk factors, this association was no longer present (adjusted hazard ratio, 1.05; 95% CI, 0.8-1.38; P =.74). CONCLUSIONS: In this low-risk population, abnormal systolic blood pressure recovery after exercise was not independently predictive of mortality after correcting for differences in baseline and exercise characteristics. PMID- 14760328 TI - Lack of aspirin effect: aspirin resistance or resistance to taking aspirin? AB - BACKGROUND: A lack of aspirin effect on platelets after a myocardial infarction (MI) is associated with poor health outcome. This lack of effect may be due to biological resistance to aspirin or due to nonadherence (the patient is not taking the aspirin, hence it has no effect). Determining which of these factors predicts poor outcome would inform potential intervention strategies. METHODS: Aspirin effect on platelets was assessed in a cohort of MI survivors who were divided into three groups: group A ("adherent"), patients whose platelets were affected by aspirin; group B ("nonadherent"), patients whose platelets showed no aspirin effect and who admitted in an interview that they were not taking their medications; and group C (potentially biologically resistant to aspirin), patients whose platelets showed no aspirin effect but maintained that they were taking their aspirin. Two health outcome measures (death, reinfarction, or rehospitalization for unstable angina; or admission for any cardiovascular causes) were assessed 12 months after enrollment. RESULTS: Seventy-three patients were enrolled and classified into groups A ("adherent," 52 patients), B ("nonadherent," 12 patients), and C ("potentially aspirin resistant," 9 patients). Adverse events and readmission were more common in the nonadherent group (B)-42% and 67%, respectively, when compared with the adherent group (A)-6% and 11%, and with the potentially biologically resistant group (C)-11% and 11%. CONCLUSIONS: Nonadherence is a significant mediator of poor outcome. It is important to evaluate whether or not patients are taking their medications in clinical settings and in studies that evaluate the effect of prescribed medications. PMID- 14760329 TI - The peripheral artery questionnaire: a new disease-specific health status measure for patients with peripheral arterial disease. AB - BACKGROUND: The most common indication for treating patients with peripheral arterial disease is to improve their health status: their symptoms, function, and quality of life. Quantifying health status requires a valid, reproducible, and sensitive disease-specific measure. The Peripheral Artery Questionnaire (PAQ) is a 20-item questionnaire developed to meet this need by quantifying patients' physical limitations, symptoms, social function, treatment satisfaction, and quality of life. METHODS: Psychometric and clinical properties of the PAQ were evaluated in a prospective cohort study of 44 patients undergoing elective percutaneous peripheral revascularization. To establish reproducibility, 2 assessments were performed 2 weeks apart and before revascularization. The change in scores before and 6 weeks after revascularization were used to determine the instruments' responsiveness and were compared with the Short Form-36 and the Walking Impairment Questionnaire. A series of cross-sectional analyses were performed to establish the construct validity of the PAQ. RESULTS: The 7 domains of the PAQ were internally reliable, with Cronbach alpha = 0.80 to 0.94. The test retest reliability analyses revealed insignificant mean changes of 0.6 to 2.3 points (P = not significant for all). Conversely, the change after revascularization ranged from 13.7 to 41.9 points (P < or =.001 for all), reflecting substantial sensitivity of the PAQ to clinical improvement. The PAQ Summary Scale was the most sensitive of all scales tested. Construct validity was established by demonstrating correlations with other measures of patient health status. CONCLUSIONS: The PAQ is a valid, reliable, and responsive disease specific measure for patients with peripheral arterial disease. It may prove to be a useful end point in clinical trials and a potential aid in disease management. PMID- 14760330 TI - Including patients with diabetes mellitus or coronary artery bypass grafting decreases the association between heart rate variability and mortality after myocardial infarction. AB - BACKGROUND: Decreased heart rate variability (HRV) is often assumed to be associated with mortality in all patients after myocardial infarction (MI), independent of clinical factors or time after MI. METHOD: HRV was determined from Holter tapes in the Cardiac Arrhythmia Suppression Trial (CAST). Patients were 71 +/- 120 days after MI. A total of 735 pre-therapy tapes were analyzed in patients who had ventricular premature contractions (VPCs) suppressed on the first treatment. The period of follow-up was 362 +/- 243 days (69 deaths). The association of clinical and demographic factors and 24-hour, daytime, and nighttime HRV to mortality in all patients, patients without coronary artery bypass graft (CABG) surgery between the qualifying MI and the Holter monitoring, and patients with neither CABG nor diabetes mellitus was determined with univariate Cox regression analysis. RESULTS: For the entire group and the subgroup without CABG, the strongest association was with increased daytime normalized high frequency power (NHF day). Further excluding patients with diabetes mellitus strengthened the association of HRV with mortality rate. Decreased natural logarithm (ln) 24-hour total and ultra low frequency (ULF) power were the strongest predictors of mortality. The best cutoff point for ln ULF for separating survivors and non-survivors was determined. After including a history of MI, congestive heart failure, or both as co-factors, ln ULF < or =7.85 identified patients at approximately 4-times the relative risk of mortality, but did not risk-stratify patients without prior MI or history of congestive heart failure. CONCLUSIONS: HRV predicts mortality rate in a broad range of times after MI. Excluding patients with CABG after MI or with diabetes mellitus significantly strengthens the association of HRV with mortality. HRV measures beyond the peri infarction period, with clinical factors, can identify subgroups at an elevated risk of mortality. PMID- 14760331 TI - Restenosis detected by routine angiographic follow-up and late mortality after coronary stent placement. AB - BACKGROUND: Routine 6-month follow-up angiography (FU angio) is the most sensitive tool to detect restenosis. Thus, FU angio protocols have been a pivotal part of trials on long-term efficacy of stents. However, it is unclear if such protocols supply data relevant for the prognosis of individual patients. The purpose of this study was to assess the impact of angiographic restenosis detected by FU angio on late mortality after coronary stent placement. METHODS AND RESULTS: We analyzed 2272 consecutive patients with successful stent placement performed from May 1992 through December 1996. All patients were scheduled for 6-month FU angio and contacted again after 4 years. FU angio was performed in 1958 patients. Of those, 557 patients (28.4%) had restenosis. After 4 years, 8.8% of patients with restenosis died, compared to 6.0% without (P =.02). There were several significant differences in clinical and angiographic characteristics between the 2 groups. In a multivariate analysis including those characteristics plus restenosis, only older age and restenosis were independent risk factors for late mortality. In patients with severe restenosis (>75% of lumen diameter; n = 231), late mortality was 7.6% in those with target vascular revascularization, compared to 14.9% without (P = not significant). CONCLUSIONS: In this analysis, mortality 4 years after stent placement was higher in patients with angiographic restenosis. Restenosis was an independent risk factor for late mortality, with a potential benefit after target vessel revascularization in severe restenoses. These data suggest that routine FU angio after stenting provides data relevant for long-term prognosis of patients. PMID- 14760332 TI - Low-dose carvedilol improves left ventricular function and reduces cardiovascular hospitalization in Japanese patients with chronic heart failure: the Multicenter Carvedilol Heart Failure Dose Assessment (MUCHA) trial. AB - BACKGROUND: The efficacy and optimum dose of beta-blockers have not been established in Japanese patients with chronic heart failure (CHF). The efficacy and safety of two doses of carvedilol, a beta-blocker with vasodilator and antioxidant actions, were investigated in Japanese patients with CHF. METHODS: After screening and a carvedilol challenge phase, 174 patients with mild to moderate CHF were randomly assigned (double-blinded) to placebo, 2.5 mg of carvedilol twice daily, or 10 mg of carvedilol twice daily. After a 2- to 4-week uptitration phase, maintenance treatment was continued for 24 to 48 weeks. The primary end point was improvement of the global assessment of CHF by the attending physician. Secondary end points were death or hospitalization for cardiovascular disease, cardiovascular hospitalization, hospitalization for heart failure, change of left ventricular ejection fraction, and change in New York Heart Association class. RESULTS: Carvedilol therapy achieved dose-dependent improvement of all end points (P for linear trend, range.002 to <.001). Both carvedilol groups showed marked risk reduction (71% to 91%) for cardiovascular and CHF hospitalization and for death or cardiovascular hospitalization (P range,.024 to <.001 for pairwise comparisons with placebo). No significant differences were observed for noncardiovascular hospitalization or adverse events. CONCLUSIONS: In Japanese patients with mild or moderate CHF, carvedilol achieved dose-related improvement of CHF and left ventricular ejection fraction; cardiovascular hospitalization was markedly reduced. At 5 mg/d, carvedilol conferred an important patient benefit, less than at 20 mg/d. PMID- 14760333 TI - Relationship between heart failure treatment and development of worsening renal function among hospitalized patients. AB - BACKGROUND: Among patients who are hospitalized with heart failure (HF), worsening renal function (WRF) is associated with worse outcomes. Whether treatment for HF contributes to WRF is unknown. In this study, we sought to assess whether acute treatment for patients who were hospitalized with HF contributes to WRF. METHODS: Data were collected in a nested case-control study on 382 subjects who were hospitalized with HF (191 patients with WRF, defined as a rise in serum creatinine level >26.5 micromol/L [0.3 mg/dL], and 191 control subjects). The association of medications, fluid intake/output, and weight with WRF was assessed. RESULTS: Calcium channel blocker (CCB) use and loop diuretic doses were higher in patients on the day before WRF (25% vs 10% for CCB; 199 +/- 195 mg vs 143 +/- 119 mg for loop diuretics; both P <.05). There were no significant differences in the fluid intake/output or weight changes in the 2 groups. Angiotensin-converting enzyme (ACE) inhibitor use was not associated with WRF. Other predictors of WRF included elevated creatinine level at admission, uncontrolled hypertension, and history of HF or diabetes mellitus. Higher hematocrit levels were associated with a lower risk. Vasodilator use was higher among patients on the day before WRF (46% vs 35%, P <.05), but was not an independent predictor in the multivariable analysis. CONCLUSIONS: Several medical strategies, including the use of CCBs and a higher dose of loop diuretics, but not ACE inhibitors, were associated with a higher risk of WRF. Although assessment of inhospital diuresis was limited, WRF could not be explained by greater fluid loss in these patients. Determining whether these interventions are responsible for WRF or are markers of higher risk requires further investigation. PMID- 14760334 TI - Effects of a home walking exercise program on functional status and symptoms in heart failure. AB - BACKGROUND: Hospital-based exercise programs using a bicycle ergometer or a combination of exercise modalities have shown positive benefits in heart failure, but may not be readily accessible to many patients. Thus, we sought to evaluate the effects of a 12-week home walking exercise program on functional status and symptoms in patients with heart failure. METHODS: A randomized controlled trial comparing a 12-week progressive home walking exercise program (n = 42) to a "usual activity" control group (n = 37) was conducted in patients with heart failure (78 [99%] male; mean age 62.6 +/- 10.6 years; ejection fraction 27% +/- 8.8%; 63 [80%] New York Heart Association class II; 15[20%] New York Heart Association class III-IV) from a Veterans Affairs medical center and a university affiliated medical center. Functional status (peak oxygen consumption via cardiopulmonary exercise testing, 6-minute walk test, the Heart Failure Functional Status Inventory), and symptoms (Dyspnea-Fatigue Index score with a postglobal rating of symptoms) were measured at baseline and 12 weeks. RESULTS: No adverse events related to exercise training occurred. Overall mean compliance to training was 74 +/- 37%. Peak oxygen consumption and the Heart Failure Functional Status Inventory were unchanged with training. Compared to the usual activity group, the training group had significantly longer walking distances measured by the 6-minute walk test (1264 +/- 255 vs 1337 +/- 272 feet, P =.001), and improved postglobal rating of symptoms (P =.03). CONCLUSION: In patients with heart failure, a progressive home walking exercise program is acceptable, increases walking distance, and decreases global rating of symptoms. PMID- 14760335 TI - Cardiac surgery for octogenarians: is it an informed decision? AB - BACKGROUND: Cardiac surgery is frequently performed to palliate cardiovascular symptoms in patients who are octogenarians, without controlled clinical trials to support its benefits. We hypothesized that death or discharge to a nursing care facility after cardiac surgery is similar in patients who are octogenarians and younger patients. METHODS: We conducted an inception cohort study in a tertiary care teaching hospital in patients who had undergone coronary grafting, valve surgery, or both over 36 months before. We collected data on preoperative disease, operation characteristics, postoperative complications, and outcome at hospital discharge. RESULTS: Of 783 patients who had cardiac surgery, 96 were octogenarians. Female sex, pulmonary hypertension, previous malignancy, cerebral vascular disease, valvular heart disease, and congestive heart failure were more frequent in patients who were octogenarians than in younger patients. Operative characteristics were similar in both age groups, except there were more frequent valve or combined with coronary grafts surgery and surgical re-exploration in octogenarians. The rate of postoperative complications including cardiovascular, neurological, renal, and nosocomial infections were higher in patients who were octogenarians than younger patients. Death or discharge to a nursing care facility was more frequent in patients who were octogenarians than younger patients (53% vs 14%, P <.002). Age > or =80 years, female sex, congestive heart failure, and surgical re-exploration were independent predictors for death or discharge to a nursing care facility after cardiac surgery. CONCLUSIONS: The rate of death or discharge to nursing care facility after cardiac surgery was high among patients who were octogenarians. Current operative outcome end points do not reflect such important differences between patients who are octogenarians and younger patients. Informed discussion of treatment options, potential for discharge to a nursing care facility, and quality of life expectations should precede a decision to undergo cardiac surgery in patients who are octogenarians. Randomized clinical trials of medical versus surgical palliation of cardiovascular symptoms in patients who are octogenarians are needed to justify cost-effectiveness and guide better use of relatively scarce Medicare resources. PMID- 14760336 TI - Peak VO2 and VE/VCO2 slope in patients with heart failure: a prognostic comparison. AB - BACKGROUND: Exercise testing with ventilatory expired gas analysis has proven to be a valuable tool for assessing patients with heart failure (HF). Peak oxygen consumption (peak VO2) continues to be considered the gold standard for assessing prognosis in HF. The minute ventilation--carbon dioxide production relationship (VE/VCO2 slope) has recently demonstrated prognostic significance in patients with HF, and in some studies, it has outperformed peak VO2. METHODS: Two hundred thirteen subjects, in whom HF was diagnosed, underwent exercise testing between April 1, 1993, and October 19, 2001. The ability of peak VO2 and VE/VCO2 slope to predict cardiac-related mortality and hospitalization was examined. RESULTS: Peak VO2 and VE/VCO2 slope were demonstrated with univariate Cox regression analysis both to be significant predictors of cardiac-related mortality and hospitalization (P <.01). Multivariate analysis revealed that peak VO2 added additional value to the VE/VCO(2) slope in predicting cardiac-related hospitalization, but not cardiac mortality. The VE/VCO2 slope was demonstrated with receiver operating characteristic curve analysis to be significantly better than peak VO2 in predicting cardiac-related mortality (P <.05). Although area under the receiver operating characteristic curve for the VE/VCO2 slope was greater than peak VO2 in predicting cardiac-related hospitalization (0.77 vs 0.73), the difference was not statistically significant (P =.14). CONCLUSIONS: These results add to the present body of knowledge supporting the use of cardiopulmonary exercise testing in HF. Consideration should be given to revising clinical guidelines to reflect the prognostic importance of the VE/VCO2 slope in addition to peak VO2. PMID- 14760337 TI - Five-year clinical and echocardiographic evaluation of the Das AngelWings atrial septal occluder. AB - BACKGROUND: The late outcome of patients treated with atrial septal occluder devices remains incompletely defined. The purpose of this study was to assess the late outcome (range 4-7 years postprocedure) of patients in whom the Das AngelWings septal occluder device was implanted in the atrial septum. We report the clinical and echocardiographic outcome, at an average of 5 years following the procedure, of patients treated with the Das AngelWings device used to close either a secundum atrial septal defect (ASD) or a patent foramen ovale (PFO). METHODS: Thirty-two patients underwent successful percutaneous closure of an atrial septal closure, patent foramen ovale, or fenestration in the lateral tunnel of their Fontan with the Das AngelWings device between June 1995 and March 1998 at Duke University Medical Center. Two of the 32 patients were lost to follow-up. The remaining 30 patients were divided into 3 groups based on indication for device implantation. Group 1 consisted of 14 patients with a secundum ASD and predominantly left-to-right atrial shunting. Group 2 consisted of 8 patients who had a PFO and who suffered a thromboembolic event. Group 3 (compassionate use) consisted of 10 patients with multiple comorbid medical problems with predominantly right-to-left shunting at the atrial level causing hypoxemia. Eight of the patients in group 3 were severely ill at the time of device implantation. The 2 remaining patients in group 3 underwent AngelWings implantation for closure of right-to-left shunting through a Fontan fenestration. Mean follow-up was 59 months. RESULTS: There was no device embolization. No patient in the ASD or PFO/stroke group had a clinical complication. By radiographic examination, 2 of 27 patients had evidence of fracture of the nitinol framework at 2-year follow-up. Residual shunting was present in 44% at 24 hours, 20% at 1 year, and 18.8% at 2 years by use of Doppler color flow imaging and/or microcavitation echocardiographic studies. Mild mitral regurgitation caused by the AngelWings device occurred in 1 patient. One patient in the compassionate use group had a subsequent neurologic event. Five of the 10 patients in the compassionate use group died of comorbid illnesses in follow-up, none directly related to device complications. CONCLUSION: The late clinical outcome of secundum ASD and PFO/stroke patients in this study demonstrates that Das AngelWings closure of the atrial septum is effective and safe. These data are encouraging with respect to the expanding use of other percutaneously implanted ASD occlusion devices, although late clinical safety and efficacy data are needed for the specific devices being implanted. PMID- 14760338 TI - How should left atrial size be reported? Comparative assessment with use of multiple echocardiographic methods. AB - BACKGROUND: Determination of left atrial (LA) size is important in clinical decision-making. The LA anteroposterior dimension (APD) has been routinely reported as LA size assessment. Early studies indicated that the APD may have limited accuracy in quantification. Conventional 3-dimensional reconstruction (C3DR) of the LA has been validated. However, its process is time-consuming and not applicable for daily practice. To explore an accurate and practical approach, we compared different echocardiographic measurements with C3DR in 141 patients with different LA sizes. METHODS AND RESULTS: LA size was measured with (1) the cubic equation with APD (Cub); (2) the ellipsoidal formula (Ellp); (3) biplane modified Simpson rule (biplane); and (4) simplified 3-dimensional reconstruction from 3 standard apical views with B spline interpolation (S3VR). All four methods were compared with C3DR. S3VR and biplane methods provided a close agreement to C3DR (y = 0.94x + 3.6, r = 0.95, SEE = 7.6 mL, mean difference = -1.3% for S3VR; y = 0.87x + 2.9, r = 0.91, SEE = 9.0 mL, mean difference = -9.4% for biplane). The Cub and Ellp calculations were less accurate, with significant volume underestimation (P <.001). CONCLUSIONS: LA single dimension is not accurate for LA size measurement. Among four different methods of LA size measurement, biplane and S3VR provide the closest agreement to C3DR. The biplane, which is readily applicable with current echocardiographic equipment, should be routinely applied in clinical practice. PMID- 14760339 TI - Red wine's antioxidants counteract acute endothelial dysfunction caused by cigarette smoking in healthy nonsmokers. AB - BACKGROUND: Long-term smoking is believed to cause endothelial dysfunction via increased oxidative stress, whereas short-term smoking impairs vasodilatation through an as yet undefined mechanism. However, red wine and its constituents have a powerful antioxidant effect both in long-term and acute consumption. The aim of the current study was to investigate whether red wine, with or without alcohol, influences endothelial dysfunction induced by acute cigarette smoking. METHODS: Sixteen healthy volunteers (8 males and 8 females) were recruited for a double-blind, crossover study, comprising 3 study days. Each subject smoked 1 cigarette, or smoked and drank 250 mL of red wine, or smoked and drank 250 mL of dealcoholized red wine. Flow-mediated dilatation (FMD) was measured after fasting and 15, 30, 60, and 90 minutes after each trial (smoke or smoke and drink either beverage). RESULTS: Acute smoking of 1 cigarette caused a reduction in FMD (P <.001), which was statistically significant 15, 30, and 60 minutes after the inhalation of smoke compared to baseline levels (P <.001, P <.001, P =.043, respectively). However, simultaneous ingestion of either red wine or dealcoholized red wine with smoking did not lead to a change in FMD. CONCLUSIONS: Acute smoking caused a significant impairment in endothelial function. Simultaneous consumption of red wine or dealcoholized red wine with smoking decreased smoke's harmful effect on endothelium. PMID- 14760340 TI - The alpha1-adrenergic blocker urapidil improves contractile function in patients 3 months after coronary stenting: a randomized, double-blinded study. AB - BACKGROUND: The recovery of left ventricular function (LVF) after revascularization takes time. alpha-Adrenergic blockade acutely improves coronary blood flow and LVF, whereas the effects of more prolonged alpha-adrenergic blockade on LVF recovery after stenting are unknown. METHODS: In 32 patients (age 58 +/- 12 y) with an 82% +/- 6% stenosis, ejection fraction (EF) and systolic thickening (%Th) were measured by transthoracic echocardiography before and 30 minutes to 2 hours after revascularization. In a double-blinded protocol, either 200 microg/kg urapidil or placebo was given intravenously, and LVF was measured 10 minutes later. Two hours later, oral treatment with 30 mg/d drug or placebo was started, and LVF measured again after 24 hours and 3 months. RESULTS: Before revascularization, EF was 49.4% +/- 8.5% (+/-SD) and 51.3% +/- 8.8% in the urapidil-treated and the placebo groups, respectively. Thirty minutes to 2 hours after coronary stenting, EF was unchanged. After intravenous drug administration, EF increased to 56.5% +/- 9.7%). At 24 hours and 3 months after revascularization, EF became 59.5% +/- 7.9% and 59.6% +/- 8.2% in the urapidil treated group, respectively, whereas EF in the placebo group did not change (50.4% +/- 5.7% and 49.7% +/- 4.9%, respectively). Revascularization did not acutely improve %Th. Intravenous urapidil improved %Th from 31.4% +/- 17.6% to 44.2% +/- 11.6%, whereas there was no change in the placebo group. At 3 months, %Th was 49.5% +/- 12.9% in the urapidil-treated group and 39.7% +/- 8.9% in the placebo group. CONCLUSIONS: These data suggest that long-term alpha-adrenergic blockade might improve LVF at midterm after coronary revascularization. PMID- 14760341 TI - Comparison between a sustained administration of carvedilol versus atenolol to reduce restenosis after coronary stenting. AB - BACKGROUND: Carvedilol is a direct inhibitor of vascular smooth muscle cell migration and proliferation through inhibition of mitogen-activated protein kinase activity and regulation of cell cycle progression. It produced an 84% suppression of neointimal hyperplasia in rat carotid angioplasty model, but no data are available regarding its effect on stent restenosis in patients. We tested whether a sustained oral administration of carvedilol reduces restenosis after coronary stenting in patients. METHODS: One hundred fifty-nine patients were randomly assigned to receive either carvedilol (50 mg/d, n = 80) or atenolol (50 mg/d, n = 79) at least 1 day before stenting and continued on the same medication over a period of 3 months. The primary end point was angiographic restenosis (>50% diameter stenosis) at follow-up angiography. RESULTS: Baseline clinical and angiographic variables were similar between the carvedilol and atenolol group. The carvedilol dose was tolerable in most patients but reduced in 3 patients because of hypotension or dizziness. Angiographic follow-up was done in 137 patients (86%), and restenosis rate was not different significantly between both groups (17.1% versus 19.4%, P =.732). CONCLUSIONS: A sustained oral administration of carvedilol is not effective to reduce restenosis after stenting in patients. With carvedilol targeting regulators of cell cycle progression and having a profound neointimal inhibition with a high blood concentration in an animal study, further investigations with a stent-based delivery to achieve a high local concentration may be warranted. PMID- 14760342 TI - Arthroscopic fixation of avulsion fractures of the tibial eminence: technique and outcome. AB - PURPOSE: The purpose of this study was to review an arthroscopic technique using screw or suture fixation for repair of types II and III fractures of the tibial eminence and review patient outcomes. TYPE OF STUDY: Retrospective review. METHODS: We conducted a review of 17 patients with Meyers and McKeever type II or III fractures of the tibial eminence treated with arthroscopic suture or screw fixation. We reviewed records and administered a questionnaire that included the International Knee Documentation Committee (IKDC) form, Tegner Activity scale, and Lysholm Knee Score. RESULTS: Five men and 12 women comprised the study group. Average age was 26.6 years (range, 7.5 to 60.1 years). Mean follow-up time was 32.6 months (range, 14 to 51 months). The study included 8 type II and 9 type III fractures. At follow-up evaluation, the mean Tegner score was 6.35 and mean Lysholm score was 94.2. In general, the best outcomes were seen in younger patients. For continuous age, significant differences were found for the IKDC functional scores, symptom scores, and IKDC final scores. For categorical age, younger patients had significantly better scores for the IKDC function and final scores. No significant differences were seen in outcomes with regard to fixation type. In 10 cases the intermeniscal ligament was interposed between the avulsed fracture and the tibia and was retracted or resected to allow fracture reduction. No significant differences were seen in the outcomes of these patients. CONCLUSIONS: We found that displaced tibial eminence fractures could be successfully treated in both younger and older patients using arthroscopic suture or screw fixation, with most patients returning to their previous activity levels. The interposed intermeniscal ligament must be retracted or resected to allow for anatomic fracture reduction. LEVEL OF EVIDENCE: Level III, Case Series. PMID- 14760343 TI - Hamstring anterior cruciate ligament reconstruction: a comparison of bioabsorbable interference screw and endobutton-post fixation. AB - PURPOSE: The purpose of this study was to evaluate hamstring anterior cruciate ligament (ACL) reconstruction using aperture fixation with bioabsorbable interference screw (BIS) and distant fixation using EndoButton (Smith & Nephew, Andover, MA) and screw-post (ENDO). TYPE OF STUDY: Prospective nonrandomized clinical outcome study. METHODS: Two groups of 15 patients who underwent autogenous hamstring ACL reconstruction with a minimum of 2 years' follow-up evaluation (mean 35 months) were included in the study. The BIS group underwent poly-L-lactic acid interference screw fixation at both femoral and tibial tunnels and the ENDO group underwent EndoButton fixation on the femoral side and screw post fixation on the tibial side. Clinical evaluation included International Knee Documentation Committee (IKDC) and arthrometer measurements. Tunnel enlargement, screw integrity, graft integrity, and graft-tunnel interface were evaluated using radiographs and magnetic resonance imaging. Statistical analyses were performed using the Student t test, with significance set at 0.05. RESULTS: All patients in both groups had functionally normal or near-normal IKDC scores. The average IKDC subjective knee form scores were 85 +/- 11 versus 81 +/- 17 (BIS v ENDO) and side to-side KT differences were 3.2 +/- 2.6 mm versus 2.4 +/- 1.8 mm (P >.05). For both groups, tunnel enlargement was present on radiographs at both femoral and tibial sides (36% to 77%). Tunnel enlargement was more significant at the femoral tunnels (53% to 77%) than the tibial tunnels (36% to 42%). In the BIS group, magnetic resonance imaging revealed 9 partially degraded screws and 2 intact screws on the femoral side. On the tibial side, 4 partially degraded and 10 intact screws were seen. For both groups, the grafts have partial or complete incorporation at all tunnels. Tunnel enlargement measured on radiographs correlate well with measurements on magnetic resonance images. CONCLUSIONS: Our results showed that rigid aperture fixation using BIS did not lead to significant differences in clinical outcome when compared with distant fixation using ENDO at 24 to 40 months follow-up evaluation. Significant tunnel enlargement was present in both groups, more pronounced on the femoral side. Magnetic resonance imaging findings showed that BIS were not degraded even at 2 to 4 years after surgery. LEVEL OF EVIDENCE: Level II. PMID- 14760344 TI - Long-term results after combined medial meniscal allograft transplantation and anterior cruciate ligament reconstruction: minimum 8.5-year follow-up study. AB - PURPOSE: The purpose of this study was to evaluate the long-term results after medial meniscal allograft transplantation combined with anterior cruciate ligament (ACL) reconstruction. TYPE OF STUDY: Retrospective clinical outcome study. METHODS: Between 1990 and 1992, 9 medial meniscal allograft transplantations were performed in symptomatic knees with a previous total or near-total medial meniscectomy. One of the 9 patients required transplant removal postoperatively because of a presumed low-grade infection versus immune reaction. The remaining 8 patients were evaluated, with an average follow-up time of 9.7 years. All 8 patients underwent an ACL reconstruction. One patient also had a staged high tibial osteotomy. RESULTS: All 8 patients were evaluated with the standard International Knee Documentation Committee (IKDC) form with no normal scores: 1 had a nearly normal score, 4 had abnormal scores, and 3 severely abnormal scores. The IKDC symptoms evaluation produced 2 normal scores, 5 nearly normal scores, and 1 abnormal score. The IKDC function test showed 5 normal scores, 1 nearly normal score, and 2 abnormal scores. Six of the 8 patients were extremely pleased with the function of the knee and were active in recreational sports. All 8 patients would recommend the procedure to a friend and would undergo the procedure again given similar circumstances. CONCLUSIONS: The findings of this study agree with several other studies with shorter follow-up times that medial meniscal allograft transplantation can significantly improve knee function in symptomatic medial meniscus-deficient knees. The addition of a ligament-stabilizing procedure probably improved the results in this patient population. LEVEL OF EVIDENCE: Level IV, Case Series. PMID- 14760345 TI - Mucoid degeneration of the anterior cruciate ligament. AB - PURPOSE: Mucoid degeneration of the anterior cruciate ligament (ACL) is not a well-known entity. Only 1 case of mucoid degeneration of the ACL has been reported in the English-language literature. This article describes 5 cases of mucoid degeneration of the ACL with clinical features, magnetic resonance imaging (MRI) findings, and a method of arthroscopic management of these cases. TYPE OF STUDY: Case series. METHODS: Over a period of 21 months from 1999 to 2001, 5 patients were diagnosed to be suffering from mucoid degeneration of the ACL using MRI, histopathologic, and arthroscopic criteria. All patients presented with progressive knee pain and restriction of flexion without history of a significant trauma or instability preceding the symptoms. MRI showed an increased signal in the substance of the ACL both in the T1- and T2-weighted images, with a mass-like configuration that was reported as a partial or complete tear of the ACL by most radiologists. At arthroscopy, the ACL was homogenous, bulbous, hypertrophied, and taut, occupying the entire intercondylar notch. The ligamentum mucosum was absent in all patients. A debulking of the ACL was performed by a judicious excision of the degenerate mucoid tissue, taking care to leave behind as much of the intact ACL as possible. Releasing it and performing a notchplasty treated impingement of the ACL to the roof and lateral wall. The ACL was not fully excised in any of the patients. RESULTS: All patients were pain free and had recovered full flexion except one who experienced painful flexion beyond 120 degrees. None of the patients showed symptoms of instability. Postoperative MRI performed after at least 12 months in 2 patients showed some intact ACL fibers in a now-thinned ACL mass. CONCLUSIONS: Mucoid degeneration of the ACL is a clinical condition afflicting active middle-aged people without a single significant traumatic episode with a specific MRI picture. They respond well to a judicious arthroscopic release of the ACL with notchplasty. LEVEL OF EVIDENCE: Level IV. PMID- 14760346 TI - Medial patellofemoral ligament reconstruction with semitendinosus autograft for chronic patellar instability: a follow-up study. AB - PURPOSE: The purpose of this study was to describe the long-term results of medial patellofemoral ligament reconstruction with a free semitendinosus graft. TYPE OF STUDY: Prospective nonrandomized study. METHODS: We assessed 15 patients (16 knees) treated between 1992 and 1996 (follow-up > 5 years). Diagnosis of patellofemoral dislocation or subluxation was based on the patient's report plus reproduction of subjective complaints of instability on physical examination. All patients were treated by the same surgeon. Assessment was performed by a different surgeon based on Crosby-Insall and Aglietti criteria. RESULTS: According to Crosby-Insall criteria, 11 knees were rated as excellent, 4 good, and 1 poor. According to Aglietti criteria, 11 knees were rated as excellent, 3 good, 1 fair, and 1 poor. According to both protocols, 15 knees showed negative apprehension test, absence of patellofemoral pain, and normal patellar tracking. In one knee, the apprehension test was positive, patellofemoral pain was present, and patellar tracking was abnormal. Patellofemoral crepitus was detected in 10 knees (attenuated in 6). Thirteen patients (14 knees) were satisfied with the results. One patient considered the result acceptable, but below expectations. The final patient underwent an additional procedure. No cases of infection and no vascular problems were seen. CONCLUSIONS: Medial patellofemoral ligament reconstruction proves to be useful in improving unstable extension mechanisms and in preventing insecurity, gradual joint deterioration, and disabling pain during dislocation episodes, in a minimal 5-year follow-up study. LEVEL OF EVIDENCE: Level IV. PMID- 14760347 TI - Arthroscopic evaluation of scaphoid waist fracture stability and the role of the radioscaphocapitate ligament. AB - PURPOSE: The purpose of this article is to arthroscopically evaluate the effect of forearm rotation on scaphoid fracture displacement and the impact of intra articular ligament sectioning. TYPE OF STUDY: Controlled laboratory study. METHODS: Scaphoid fracture stability is studied arthroscopically in 10 cadaveric upper limbs. Displacement of the osteotomized scaphoid with and without forearm rotation is arthroscopically evaluated before and after radioscaphocapitate (RSC) ligament sectioning. RESULTS: No rotation at the fracture site was identified in full pronation and full supination with the wrist immobilized. With the RSC ligament intact, no immobilization, and the wrist fully pronated, 25% of scaphoid fractures rotated less than 1 mm, 62.5% rotated 1 to 2 mm, and 12.5% rotated more than 2 mm. After sectioning the RSC ligament, the fully pronated wrist resulted in rotation of less than 1 mm in 75% and 1 to 2 mm in 25%. No rotation at the fracture site was seen with supination, with or without immobilization. No difference was found between loaded and unloaded trials. CONCLUSIONS: Based on this observational data, it appears safe to use a below-elbow thumb spica cast in the treatment of minimally displaced scaphoid waist fractures. Sectioning of the RSC ligament resulted in reduced amount of rotation at the scaphoid waist fracture; thus the RSC ligament may be a deforming force rather than a stabilizing force in scaphoid waist fractures. Arthroscopy may be a valuable tool in the study of the effect of ligament sectioning on fracture stability. PMID- 14760348 TI - The use of a volar ulnar portal in wrist arthroscopy. AB - PURPOSE: The purpose of this study was to quantitatively describe the neurovascular relationships of a volar ulnar (VU) wrist arthroscopy portal. A second purpose was to evaluate whether the use of a VU portal identified additional pathology of the lunotriquetral ligament and ulnar sling mechanism that was not seen through the dorsal portals. TYPE OF STUDY: This study was an anatomic study and retrospective chart review. METHODS: Cadaver dissections established the neurovascular anatomy of the VU portal. Measurements were taken from the portal to the ulnar nerve and artery, the palmar cutaneous branch of the ulnar nerve, and the pronator quadratus. A dorsal capsulotomy was performed to assess the ligamentous interval. A chart review was performed of 23 patients in whom a volar ulnar portal was used. Intraoperative pathology that was identified through the VU portal but was not visible through a dorsal portal was recorded. Postoperative neurovascular complications were noted. RESULTS: The portal was generally > 5 mm radial to the ulnar neurovascular bundle, but no true internervous plane was seen. Tears of the palmar aspect of the lunotriquetral ligament were seen in 7 patients. One patient had a triangular fibrocartilage tear that extended into the dorsal radioulnar ligament. CONCLUSIONS: This study provides a safe, standardized approach to the volar ulnar aspects of the radiocarpal joint, which is useful for evaluation of the ulnar sling mechanism and the dorsal radioulnar ligament. The VU portal aids in the diagnosis and debridement of tears involving the palmar aspect of the lunotriquetral ligament. The VU portal should be considered for inclusion in the arthroscopic examination of any patient with ulnar sided wrist pain. LEVEL OF EVIDENCE: Level IV. PMID- 14760349 TI - Normal variations of the glenohumeral ligament complex: an anatomic study for arthroscopic Bankart repair. AB - PURPOSE: The purpose of this study was to evaluate the prevalence and morphologic characteristics of normal variations in the glenohumeral ligament complex (GHLC). TYPE OF STUDY: Anatomic study in cadavers. METHODS: We investigated 84 cadaver shoulders and recorded visualization of the superior glenohumeral ligament (SGHL), middle glenohumeral ligament (MGHL), and anterior band of the inferior glenohumeral ligament (AIGHL). In cases with an AIGHL, we noted the location of its origin with respect to the anterior glenoid in terms of a clock face as in a right shoulder. If an MGHL was present, the site of its glenoid attachment was recorded. The presence of a cord-like MGHL, the Buford complex, and a sublabral foramen was also investigated. RESULTS: Of these, 79 (94.1%) manifested an SGHL, 53 (63.1%) an MGHL, and 76 (90.5%) an AIGHL. The AIGHL originated in an area located between the 2- and 5-o'clock position; in 11 (14.5%), the origin was at the 2-o'clock position; in 49 (64.5%) at the 3-o'clock position; in 11 (14.5%), the 4-o'clock position; and in 5 (6.5%) at the 5-o'clock position. Two common variations in the attachment of the MGHL were seen; 30 of 53 MGHL (56.6%) originated from the labrum separate from the origin of the SGHL, and 23 (43.4%) from the labrum at the origin of the SGHL. Of the 84 specimens, 15 (17.9%) manifested a cord-like MGHL, and one (1.2%) the Buford complex. None of the specimens had a sublabral foramen, a finding that requires further investigation. CONCLUSIONS: Our results suggest that the Buford complex is a rare variant of the GHLC, and the cord-like MGHL appears to be a relatively common normal variant. CLINICAL RELEVANCE: The present study provides useful information concerning normal variations of the GHLC to arthroscopists considering Bankart repair. PMID- 14760350 TI - The inverted pear glenoid: an indicator of significant glenoid bone loss. AB - PURPOSE: The purpose of this study was to determine the amount of glenoid bone loss required to produce an inverted pear glenoid. TYPE OF STUDY: Two-part anatomic study involving live subjects and cadaveric specimens. METHODS: From June 2000 to April 2002, 53 patients (mean age, 30 +/- 13 years) underwent arthroscopic evaluation and treatment with a diagnosis of anterior instability by the senior author (S.S.B.). Each of these patients underwent a 3-portal diagnostic arthroscopy to determine the morphology of the glenoid. Glenoids were classified as either inverted pear or non-inverted pear glenoids based on the visual appearance. The amount of glenoid bone loss was then quantified arthroscopically using a validated methodology. In addition, 6 fresh-frozen cadaveric specimens (mean age, 74.1 +/- 7.4 years) were dissected and evaluated to determine the minimum amount of bone loss required to produce an inverted pear glenoid. RESULTS: Forty-two patients were classified as having non-inverted pear glenoids and 11 patients were classified as having inverted pear glenoids. The mean amount of bone loss anteriorly was significantly more (P <.000006) in the inverted pear glenoid group (mean, 8.6 mm; range, 6 to 12 mm) than the non inverted pear glenoid group (mean, 1.5 mm; range, 0 to 3.0 mm). The percentage of loss of glenoid width was also significantly different (inverted pear mean, 36%; range, 25% to 45% versus non-inverted pear mean, 6.2%; range, 0% to 12.5%; P <.000006). The mean amount of bone loss required in cadaveric specimens to convert a normal pear-shaped glenoid into an inverted pear glenoid was 7.5 mm (range, 6.5 to 9.0 mm), representing 28.8% of the glenoid width (range, 27% to 30%). CONCLUSIONS: The results of this study show that the majority of patients with the diagnosis of anterior instability show some bone loss anteriorly. However, the inverted pear glenoid represents a significant amount of bone loss, at least 25% to 27% of the width of the inferior glenoid. In patients with an inverted pear glenoid, a bone grafting procedure to restore the normal articular arc of the glenoid should be strongly considered to re-establish normal stability to the shoulder. PMID- 14760351 TI - Variations in glenoid rim anatomy: implications regarding anchor insertion. AB - PURPOSE: The purpose of this study was to investigate normal bony anatomy of the glenoid rim and to define the angles for successful anchor placement for anterior and posterior labral repairs. TYPE OF STUDY: An anatomic study using cadaveric shoulder specimens. METHODS: Soft tissue was dissected from 20 cadaveric shoulders, and the glenoids were isolated. The glenoid specimens were scanned to obtain cross-sectional images using peripheral quantitative computed tomography (pQCT) in 4 different planes. Glenoid rim angles were measured from the cross sectional pQCT images of the glenoids at 5 positions: the 3-o'clock, 4:30-, 6-, 7:30-, and 9-o'clock positions. Glenoid morphology was noted for each position. RESULTS: The glenoid rim angles from the 3-o'clock to the 9-o'clock positions were 53 degrees +/- 5 degrees, 45 degrees +/- 7 degrees, 80 degrees +/- 10 degrees, 61 degrees +/- 10 degrees, 49 degrees +/- 4 degrees, respectively. Asymmetric morphology of the glenoid was noted with an almost straight line extending medially from the rim at the 3-o'clock position, whereas a concave morphology was noted at the 9-o'clock position. Similarly at the 4:30-o'clock position, the scapular bony surface did not curve toward the base as markedly as it did at the corresponding posterior 7:30-o'clock position. CONCLUSIONS: The available bone mass for the anchor insertion was found to vary depending on the position of the glenoid rim. Both rim angle and glenoid morphology for each position must be considered when selecting the ideal anchor insertion angle for labral repair. PMID- 14760352 TI - Glenohumeral volume reduction in arthroscopic shoulder reconstruction: a cadaveric analysis of suture plication and thermal capsulorrhaphy. AB - PURPOSE: The purpose of this study was to objectively assess glenohumeral volume changes after arthroscopic suture plication, arthroscopic thermal capsulorrhaphy, and combined suture and thermal treatment. TYPE OF STUDY: Cadaveric study. METHODS: Five matched pairs of cadaver shoulders were treated with either thermal capsulorrhaphy (n = 5) or arthroscopic suture plication (n = 5). The shoulders treated with plication were then treated with thermal capsulorrhaphy to form a combined treatment group (plication and thermal). Preoperative and postoperative glenohumeral volume measurements were obtained, and the techniques were statistically compared with a 1-factor analysis of variance. RESULTS: All 3 procedures produced marked decreases in glenohumeral capsular volume. Suture plication reduced intra-articular volume an average of 19.0%. Thermal capsulorrhaphy resulted in a mean capsular volume reduction of 33.4%. Combined treatment with suture plication and thermal capsulorrhaphy reduced glenohumeral volume an average of 41.0%. Both thermal capsulorrhaphy and the combined treatment produced significantly greater reduction in glenohumeral volume than plication alone (P <.0001). Although combined suture plication and thermal capsulorrhaphy resulted in greater reduction in capsular volume than thermal capsulorrhaphy alone, the differences were not statistically significant. CONCLUSIONS: Our results indicate that arthroscopic suture plication and thermal capsulorrhaphy are both effective in reducing glenohumeral intra-articular volume. Thermal capsulorrhaphy alone or in combination with suture plication led to significantly greater reduction in capsular volume when compared with isolated suture plication. PMID- 14760353 TI - Influence of test temperature and test speed on the mechanical strength of absorbable suture anchors. AB - PURPOSE: Absorbable implant materials offer various advantages but are mechanically far weaker than metals. Despite known temperature dependence of the biomechanical properties of these materials, mechanical testing has almost exclusively been performed at room temperature in the literature. In this study, the difference in mechanical performance at room and body temperature was assessed in vitro at different test speeds. TYPE OF STUDY: Biomechanical bench study. METHODS: Five absorbable suture anchor models were held in a metallic holder and loaded under tension using 0.5-mm steel wires until failure. Testing temperature was 20 degrees C +/- 1 degrees C or 37 degrees C +/- 1 degrees C, test speed was 50 mm/min or 5 mm/min. Tensile load at failure and failure mode were recorded. To test creep behavior, a constant load of 100 N was applied, and time to failure was recorded at both temperatures. RESULTS: Both raising the temperature and decreasing test speed significantly (P <.0001) impaired the mechanical performance of the tested implants. Increase of temperature (20 degrees C to 37 degrees C) resulted in a decrease of the maximal failure strength by up to 40% and decreased time to failure by up to 98% under static load. At 37 degrees, decreasing the test speed from 50 to 5 mm/min lowered the load to failure by up to 18%. Failure of the anchors always occurred by eyelet cutout of the wire. CONCLUSIONS: The lower the test speed, the higher is the influence of the testing temperature. Testing of implants at room temperature instead of body temperature may falsely improve test results by a factor of up to 50 under static load. Therefore, testing absorbable implants at body temperature seems mandatory, preferably at slow test speeds. PMID- 14760354 TI - Locked knee during pregnancy. AB - An acutely locked knee is a painful and debilitating orthopaedic condition usually caused by a loose body or bucket-handle meniscal tear that requires surgery to remove or repair. We describe 2 cases of acute locking of the knee that occurred during pregnancy. Both patients underwent urgent arthroscopic treatment under spinal anesthesia without complication to the patient or fetus. Concerns regarding surgery during pregnancy revolve around the safety for the mother and fetus. Although local and regional anesthetics are safe with proper management, there is a slightly increased risk of spontaneous abortion when general anesthesia is used, especially in the first trimester. Although purely elective surgery is never recommended during pregnancy, arthroscopic knee surgery can be performed safely in emergency situations with proper planning by a coordinated team including the surgeon, anesthesiologist, and obstetrician. PMID- 14760355 TI - Repeated hemarthrosis with massive rotator cuff tear. AB - An 80-year-old woman with repeated hemarthrosis of the right shoulder was examined and treated arthroscopically. Plain radiographs indicated narrowing of the acromiohumeral interval (3 mm) with large acromial spur. Under fluoroscopic visualization, the impingement between the humeral head and the medial edge of the acromion was seen at 50 degrees abduction. The area of impingement of the humeral head seemed to be just medial to the greater tuberosity. Arthroscopically, a crater formation was seen at this site. Active bleeding was seen at the center of the crater, which was interpreted as the principal cause of intra-articular hemorrhaging. The undersurface of the acromion was irregular because of the presence of osteophyte. At 50 degrees abduction, the crater impinged with the medial edge of the acromion. We assume that both the presence of osteophyte and the instability caused by the massive rotator cuff tear might contribute to the attrition of the humeral head. In this patient, intra-articular hemorrhaging was successfully treated arthroscopically by coagulation of the bleeding point with minimal abrasion of the undersurface of the acromion. We believe that the arthroscopy facilitated the clinical management of the repeated hemarthrosis with massive rotator cuff tear. PMID- 14760356 TI - Arthroscopically assisted replacement of dynamic hip screw for unrecognized joint penetration of lag screw through a new portal. AB - Nails penetrating into the hip joint after fixation of intertrochanteric fractures may account for one third of all treatment failures. Some authors recommend leaving the nail in the penetrated position until union is certain. However, if the lag screw is long enough penetrate the acetabular joint surface and cause severe pain and limited range of motion, a new shorter one should replace it. In this situation, direct visualization of the joint surface may not be possible, and fluoroscopy can be difficult to interpret. To our knowledge, this is the first case report of left intertrochanteric fracture with unrecognized joint penetration of the lag screw that was replaced by a new shorter one with arthroscopic assistance through a new portal of the screw canal. This case constitutes a unique and interesting application of hip arthroscopy. PMID- 14760357 TI - Arthroscopic biceps tenodesis. AB - Arthroscopic biceps tenodesis is indicated for the treatment of severe biceps tendonopathy, partial- or full-thickness tendon tears, or biceps instability typically associated with rotator cuff tear, although there has been considerable debate on tenotomy versus tenodesis. We advocate tenodesis, for the following reasons: to re-establish the resting muscle length so as to avoid scaring and spasm, to allow biceps use for complex elbow motion, and to avoid cosmetic defects in cases in which deformity can sometimes equal disability. This technical note provides illustrations and detailed descriptions of our arthroscopic tenodesis technique using a Arthrex (Naples, FL) biotenodesis system. PMID- 14760358 TI - Transtendon arthroscopic repair of partial-thickness, articular surface tears of the rotator cuff. AB - Partial tears of the articular surface of the rotator cuff are common lesions and when not repaired can lead to persistent pain and disability. In this report, we describe a transtendon method of repairing partial articular surface lesions that anatomically restores the footprint of the rotator cuff, allowing a broad area for tendon healing to bone. PMID- 14760362 TI - Sheltered load associated with S-alleles in Solanum carolinense. AB - Bud pollinations allowed me to examine the effects of homozygosity at loci in the area of suppressed recombination around the S-locus in Solanum carolinense, whose S-alleles show surprisingly low diversification rates. The total number of seeds produced was lower for incompatible than compatible pollinations, revealing that self-incompatibility was only somewhat overcome by bud pollination. However, low seed set in incompatible crosses was not due solely to the incompatibility response; crosses between distinct plants sharing the same alleles at the S-locus led to dramatically high seed abortion, nearly equal to that found upon selfing. An excess of heterozygotes in the surviving progeny supports the supposition that these high abortion rates are due to sheltered load, that is, previously unexpressed load accumulated due to enforced heterozygosity and recombination suppression around the S-locus. Of the seven alleles examined in total, two showed evidence of severe load and five did not. The magnitude of load was consistent with terminal branch length in some, but not all, cases. PMID- 14760363 TI - Vlax Roma history: what do coalescent-based methods tell us? AB - Three coalescent-based methods allowed us to infer some aspects of the history of three Bulgarian Gypsies populations belonging to the Vlax linguistic group: the Lom, Rudari and Kalderas. We used several kinds of genetic markers: HV1 sequences of the maternally inherited mitochondrial genome and microsatellites of the paternally inherited Y chromosome and of the biparentally inherited chromosome 8. This allowed us to infer several parameters for men and women: the splitting order of the populations and the ages of the splitting events, the growth rate in each population and the migration rates between populations. Altogether, they enabled us to infer a demographic scenario that could explain the genetic diversity of Vlax Roma: recent splits occurring after the arrival in Europe, asymmetric migration flows especially for males and unequal growth rates. This represents a considerable contribution to the Vlax Roma history in comparison with the inferences from classical population genetics. PMID- 14760364 TI - Integrins: roles in cancer development and as treatment targets. AB - The integrin family of cell adhesion proteins promotes the attachment and migration of cells on the surrounding extracellular matrix (ECM). Through signals transduced upon integrin ligation by ECM proteins or immunoglobulin superfamily molecules, this family of proteins plays key roles in regulating tumour growth and metastasis as well as tumour angiogenesis. Several integrins play key roles in promoting tumour angiogenesis and tumour metastasis. Antagonists of several integrins (alpha5beta1, alphavbeta3 and alphavbeta5) are now under evaluation in clinical trials to determine their potential as therapeutics for cancer and other diseases. PMID- 14760365 TI - Gefitinib ('Iressa', ZD1839) and new epidermal growth factor receptor inhibitors. AB - The epidermal growth factor receptor (EGFR) is a promising target for cancer therapy and a number of EGFR-targeted agents have been developed. Those most advanced in development are the EGFR tyrosine kinase inhibitors gefitinib ('Iressa', ZD1839) and erlotinib ('Tarceva', OSI-774), and the monoclonal antibody cetuximab ('Erbitux', IMC-C225). This review provides a clinical overview of these agents, highlighting their antitumour activities in different tumour types. Epidermal growth factor receptor-targeted agents are generally well tolerated and are not typically associated with the severe adverse events often seen with cytotoxic chemotherapy. Gefitinib is the agent with the most extensive clinical experience, particularly in non-small-cell lung cancer (NSCLC). Recently, gefitinib became the first-approved EGFR-targeted agent, for use in patients with previously treated advanced NSCLC in Japan, the USA and other countries. Further studies are required to explore the full potential of these novel agents either as monotherapy or combination therapy. PMID- 14760366 TI - Control of oncogenesis and cancer therapy resistance. AB - Despite the combined action of surgery, radiotherapy and chemotherapy, the leading cause of death in cancer patients continues to be the acquired, or intrinsic, tumour resistance to therapy. Some of the genetic alterations that contribute to the malignant transformation are involved in maintaining cell survival under uncontrolled growth conditions. Chemotherapy agents, as well as radiotherapy, trigger a series of signalling pathways in the cells that activate not only the apoptotic machinery, but also cell-survival pathways. In this scenario, the efficacy of therapy is the result of balance between the apoptotic and the survival pathways activated in the tumour, and those elicited by the therapeutic agent. Apoptosis is one of the programmes usually altered in most cancers so as to guarantee tumour progression and, often, these alterations are responsible for therapy resistance, as well. PMID- 14760367 TI - Selective bladder preservation for muscle-invasive transitional cell carcinoma of the urinary bladder. AB - Invasive transitional cell carcinoma (TCC) of the urinary bladder is traditionally treated with radical cystectomy. This approach results in great morbidity and lifestyle changes, and approximately half of the patients treated in this way will experience recurrent TCC despite surgery. An alternative approach using selective bladder-preservation techniques incorporates transurethral resection of bladder tumours, radiation therapy, and chemotherapy. Over the past 20 years, international experience has demonstrated that this approach is feasible, safe, and well tolerated. Furthermore, the long-term outcomes of overall survival and disease-free survival compare favourably with the outcomes from radical cystectomy. The most important predictor of response is stage, with significantly higher long-term survival in patients with T2 disease. Another important positive predictor of complete response to therapy is the ability of the urologic oncologist to remove all visible tumour through a transurethral approach prior to initiation of radiation therapy. A negative predictive factor is the presence of hydronephrosis, and age and gender do not affect disease-free survival. The majority of patients who enjoy long-term survival do so with an intact native bladder. Quality of life studies have demonstrated that the retained bladder functions well in nearly all of these patients. Selective bladder preservation will not entirely take the place of radical cystectomy, but should be offered as an important alternative to patients newly diagnosed with muscle-invasive TCC. PMID- 14760368 TI - Patient satisfaction with two different models of cancer genetic services in south-east Scotland. AB - There is a need to integrate primary- and secondary-care cancer genetic services, but the most appropriate model of service delivery remains unclear. This study reports patients' expectations of breast cancer genetic services and a comparison of their satisfaction with two service models. In the first model, risk assessment was carried out using mailed family history data. Women estimated as being at high/moderate risk were offered an appointment at the familial breast cancer clinic, and those at low risk were sent a letter of reassurance. In the second model, all women were seen by a genetic nurse specialist, who assessed risk, referred high/moderate-risk women to the above clinic and discharged those at low risk. Over 60% of all women in the study regarded access to breast screening by mammogram and regular check-ups as very important. This underlines the demand for a multidisciplinary service providing both clinical genetic and surgical services. Satisfaction was high with both models of service, although significantly lower among women not at increased cancer risk and thus not offered a clinical check-up and mammography. Increased cancer worry was associated with a greater expressed need for information and for reassurance through follow-up clinical checks and mammography. Better targeting of counselling to the expressed concerns and needs of these women is required to improve the service offered. GPs and patients expressed no clear preference for any specific service location or staffing configuration. The novel community service was less expensive in terms of both staff and patient costs. The potential to decrease health staff/patient contact time and to employ nurse practitioners with both clinical genetic and oncology training should be explored further. The rapidly rising demand for these services suggests that the evaluation of further new models needs to continue to be given priority to guide the development of cancer genetic services. PMID- 14760369 TI - Clinical and endocrine data for goserelin plus anastrozole as second-line endocrine therapy for premenopausal advanced breast cancer. AB - A total of 16 premenopausal women with metastatic breast cancer (N=13) or locally advanced primary breast cancer (N=3) were treated with a combination of a gonadotropin-releasing hormone agonist goserelin, and a selective aromatase inhibitor anastrozole. All had previously been treated with goserelin and tamoxifen. In all, 12 patients (75%) achieved objective response or durable stable disease at 6 months, with a median duration of remission of 17+ months (range 6-47 months). Four patients still have clinical benefit. Introduction of goserelin and tamoxifen resulted in an 89% reduction in mean oestradiol levels (pretreatment vs 6 months=224 vs 24 pmol l(-1)) (P<0.0001). Substitution of tamoxifen by anastrozole on progression resulted in a further 76% fall (to 6 pmol l(-1) at 3 months) (P<0.0001). Treatment with goserelin and tamoxifen led to a 90% fall in the mean follicle-stimulating hormone (P<0.001). This was reversed once therapy was changed to goserelin and anastrozole. A similar initial reduction was seen in the mean luteinising hormone levels, but substitution of tamoxifen by anastrozole on progression resulted in no significant change. Goserelin and tamoxifen did not lead to any significant change in testosterone and androstenedione levels. The combined use of goserelin and anastrozole as second-line endocrine therapy produces a significant clinical response of worthwhile duration, with demonstrable endocrine changes, in premenopausal women with advanced breast cancer, and offers them another therapeutic option. Further studies involving more patients and longer follow-up are indicated. PMID- 14760370 TI - Diagnostic accuracy for different strategies of image-guided breast intervention in cases of nonpalpable breast lesions. AB - To find out whether ultrasound-guided fine-needle aspiration (FNA) and ultrasound and stereotactic-guided large core needle biopsy (LCNB) are reliable alternatives to needle-localised open breast biopsy (NLBB) in daily practice, we performed a retrospective study and evaluated the validity of these methods. In all, 718 women with 749 nonpalpable breast lesions from three Dutch Hospitals were included, and the validity of the various methods for diagnosis was assessed. This was carried out according to a method described by Burbank and Parker for evaluating the quality of an image-guided breast intervention. We compared our results with the outcome of the COBRA study. Overall, all diagnostic strategies (NLBB, FNA, LCNB ultrasound and stereotactic guided) show comparable agreement rates. However, the miss rates differ: 2% for NLBB, 3% for COBRA (LCNB in study setting), 5% for FNA and 8-12% for LCNB in practice. Fine-needle aspiration was nonconclusive in 29%, and shows an overestimation for DCIS in 9%. The DCIS underestimate rate in NLBB was 8%. For the assessment of lesions consisting of microcalcifications only and to exclude malignancy in all other lesions, a 14 gauge needle should be used. Ultrasound-guided intervention can be performed in a large percentage of nonpalpable lesions. Lesions consisting only of microcalcifications on mammography need special attention. PMID- 14760371 TI - Bleomycin, vincristine, cisplatin/bleomycin, etoposide, cisplatin chemotherapy: an alternating, dose intense regimen producing promising results in untreated patients with intermediate or poor prognosis malignant germ-cell tumours. AB - Patients with poor and intermediate prognosis metastatic germ-cell tumours (MGCTs) are at a significant risk of relapse after standard platinum-based chemotherapy. Novel treatment regimens are required to improve survival. Dose intense, alternating combinations of drugs with known activity in germ-cell tumours represents one approach. In all, 43 patients with IGCCCG intermediate/poor prognosis MGCT were treated with a dose intense regimen alternating bleomycin, vincristine, cisplatin (BOP) with bleomycin, etoposide, cisplatin (BEP) to a maximum of three cycles. Data were collected on the maintenance of dose intensity, toxicity, response, progression-free (PFS) and overall survival (OS). The complete response rate was 58%; a further 7% of patients being rendered disease free by resection of viable residual tumour. With a median follow-up of more than 4 years in surviving patients, 3-year OS and PFS rates of 81% (95% CI: 66-91%) and 72% (95% CI: 56-83%) are seen, respectively. Bleomycin, vincristine, cisplatin (BOP)/bleomycin, etoposide, cisplatin (BEP) was well tolerated, with 86% of patients completing all planned courses. Toxicity was predominantly haematological with common toxicity criteria grade III neutropenia in 90% of patients. Cisplatin neuropathy and bleomycin-induced pulmonary toxicity represented the most significant nonhaematological toxicity. Bleomycin, vincristine, cisplatin (BOP)/bleomycin, etoposide, cisplatin (BEP) represents a practicable, well-tolerated, dose intense chemotherapy regimen with significant activity in intermediate and poor prognosis MGCT. PMID- 14760372 TI - Increased mortality rates in young and middle-aged patients with malignant germ cell tumours. AB - Cisplatin-based chemotherapy of malignant germ cell tumours (MGCT) has been reported to increase the risk of cardiovascular morbidity. A high incidence of second nongerm cell malignancies is well documented in MGCT survivors. The death risk due to these conditions is, however, more unknown in MGCT patients. Standard mortality rates (SMRs) were established in 3378 Norwegian MGCT patients treated from 1962 to 1997 aged 10.6% (P=0.016). Baseline or change in Ki-67 LI did not correlate to survival. These data suggest that IFN alpha in vivo has only modest effect on tumour proliferation in patients with mRCC. Tumour Ki-67 (MIB-1) reactivity after 1 month of immunotherapy appears to be a significant predictor of patient survival. PMID- 14760376 TI - Serum bile acids and risk factors for colorectal cancer. AB - The ratio of deoxycholic acid to chenodeoxycholic acid in the serum of 62 men was inversely related to body mass index and to saturated fat intake after adjustment for body mass index, smoking, and age conversely, this ratio was associated positively with the intake of fibre from grains. PMID- 14760377 TI - Statin use and cancer risk in the General Practice Research Database. AB - In a matched case-control study using the General Practice Research Database, current statin use was not associated with a significantly altered risk of any of 13 studied cancers. Untreated hyperlipidaemia was associated with slightly increased risks of colon cancer (relative risk 1.8; 95% confidence interval 1.2 2.8), prostate cancer (1.5; 1.1-2.0), and bladder cancer (1.9; 1.2-3.1). PMID- 14760378 TI - Prevalence of papillomavirus infection in women in Ibadan, Nigeria: a population based study. AB - To investigate the prevalence of and the risk factors for cervical infection with human papillomavirus (HPV) in an inner-city area of Ibadan, Nigeria, we interviewed and obtained a sample of cervical cells from 932 sexually active women aged 15 years or older. A total of 32 different HPV types were identified with an HPV prevalence of 26.3% overall and 24.8% among women without cervical lesions; or age-standardised to the world standard population of 28.3 and 27.3%, respectively. High-risk HPV types predominated, most notably HPV 16, 31, 35 and 58. In all, 33.5% of infections involved more than one HPV type. Unlike most populations studied so far, HPV prevalence was high not only among young women, but also in middle and old age. Single women (odds ratio, OR=2.1; 95% confidence interval, CI=1.1-3.9) and illiterate women (OR=1.7; 95%CI=1.1-2.5) showed increased HPV positivity. Associations were also found with anti-Herpes simplex-2 antibodies (OR=1.6; 95% CI: 1.1-2.1) and with the husband's extramarital relationships (OR=1.6: 95% CI: 1.0-2.6). High prevalence of HPV in all age groups may be a distinctive feature of populations where HPV transmission continues into middle age and cervical cancer incidence is very high. PMID- 14760379 TI - Filter cigarette smoking and lung cancer risk; a hospital-based case--control study in Japan. AB - Recent changes in the histology of lung cancer, namely a relative increase of adenocarcinoma compared to squamous cell carcinoma, might be due to a temporal shift from nonfilter to filter cigarettes. To investigate the association between type of cigarette and lung cancer by histological type, we conducted a case control study in Japan, comprising 356 histologically confirmed lung cancer cases and 162 controls of male current smokers, who provided complete smoking histories. Overall, logistic regression analysis after controlling for age and prefecture revealed decreased risk, as shown by adjusted odds ratios, for both squamous cell carcinoma and adenocarcinoma among lifelong filter-exclusive smokers as compared to nonfilter or mixed smokers. This decrease was greater for squamous cell carcinoma than for adenocarcinoma. Among men under 54 years, filter exclusive smokers displayed increased risk of adenocarcinoma, but decreased risk of squamous cell carcinoma. The recent shift in histology from squamous cell carcinoma to adenocarcinoma, particularly among younger smokers, might be due to changes in cigarette type. However, among subjects aged 65 years or more, no differences in histological type appeared related to type of cigarette smoked, implying that other factors are associated with increases in adenocarcinoma among older Japanese population. PMID- 14760380 TI - Declining mortality from breast cancer in Yorkshire, 1983-1998: extent and causes. AB - Incidence and mortality data, stage of disease and treatment information for female breast cancer were obtained for the years 1975-1999 for the former Yorkshire Regional Health Authority area, from the Northern and Yorkshire Cancer Registry and Information Service. Deaths by age group and 3-year calendar period were separated into those occurring among cases diagnosed 0-2 and 3-4 years preceding death (short- and medium-term survivors, respectively), and among longer-term survivors and age-adjusted rates were calculated separately by survival time. The 3-year survival of cases incident in 1991-1999 and in 1982 1990 were compared, adjusting for stage and treatment. Breast cancer mortality in Yorkshire stopped increasing around 1983 and has since consistently declined, primarily among short- and medium-term survivors. The 3-year survival was significantly improved in cases diagnosed in 1991-1999, compared with 1982-1990, in all age groups (hazard ratio (HR)=0.75, 95% c.l. 0.71-0.78). The improvement was least in the 65+ years age group (HR=0.83, 95% c.l. 0.79-0.88), intermediate in the youngest (<50 years) cases (HR=0.71, 95% c.l. 0.63-0.80) and greatest in the age group 50-64 years, offered routine screening after 1988 (HR=0.51, 95% c.l. 0.47-0.57). The benefit for cases diagnosed in the 1990s persisted, but was reduced after adjustment for stage in cases <65 years, while it disappeared in older cases (HR=1.01). Below age 65 years, the use of systemic therapy increased substantially, but did not explain the residual improvements in short-term prognosis. A greater decline in breast cancer mortality in Yorkshire from 1982 to 1984 was observed among short- and medium-term, than in longer-term, survivors. Much of the improvement in survival in cases <65 years could be attributed to a more favourable stage at diagnosis, whereas this accounted for nearly all the improvement in survival among older cases. Systemic therapy had little or no impact on 3-year survival. Continuing declines in mortality can be expected in the current decade, as a result of the long-term effects of both mammography screening and increased use of systemic therapy. PMID- 14760381 TI - MUC4 mucin expression in human pancreatic tumours is affected by organ environment: the possible role of TGFbeta2. AB - MUC4 is highly expressed in human pancreatic tumours and pancreatic tumour cell lines, but is minimally or not expressed in normal pancreas or chronic pancreatitis. Here, we investigated the aberrant regulation of MUC4 expression in vivo using clonal human pancreatic tumour cells (CD18/HPAF) grown either orthotopically in the pancreas (OT) or ectopically in subcutaneous tissue (SC) in the nude mice. Histological examination of the OT and SC tumours showed moderately differentiated and anaplastic morphology, respectively. The OT tumour cells showed metastases to distant lymph nodes and faster tumour growth (P<0.01) compared to the SC tumours. The MUC4 transcripts in OT tumours were very high compared to the undetectable levels in SC tumours. The SC tumour cells regained their ability to express MUC4 transcripts after in vitro culture. Immunohistochemical analysis using MUC4-specific polyclonal antiserum confirmed the results obtained by Northern blot analysis. Interestingly, the OT tumours showed expression of TGFbeta2 compared to no expression in SC, suggesting a possible link between MUC4 and TGFbeta2. The MUC4 expression, morphology, and metastasis of human pancreatic tumour cells are regulated by a local host microenvironment. TGFbeta2 may serve as an interim regulator of this function. PMID- 14760382 TI - Overexpression of dopa decarboxylase in peritoneal dissemination of gastric cancer and its potential as a novel marker for the detection of peritoneal micrometastases with real-time RT-PCR. AB - We previously performed a global analysis of the gene expression of gastric cancer cell lines established from metastases to the peritoneal cavity with the cDNA microarray method, which made it possible to analyse the expression of approximately 21168 genes for the identification of novel markers for the detection of micrometastases in the peritoneal cavity. One of the upregulated genes is dopa decarboxylase (DDC), which is responsible for the synthesis of the key neurotransmitters dopamine and serotonine. We have examined its potential as a novel marker for the detection of peritoneal micrometastases of gastric cancer.DDC mRNA in the peritoneal wash from 112 gastric cancer patients was quantified for comparison of carcinoembryonic antigen (CEA) mRNA by means of real time reverse transcriptase-polymerase chain reaction (RT-PCR) with a fluorescently labelled probe to predict peritoneal recurrence. The quantity of DDC and CEA correlated with wall penetration. Real-time RT-PCR could quantitate 10-10(6) DDC-expressing gastric cancer cells per 10(7) mesothelial cells. The cutoff value was set at the upper limit of the quantitative value for noncancer patients, and those above this cutoff value constituted the micrometastasis (MM+) group. Of 15 cases with peritoneal dissemination, 13 were MM+DDC (87% sensitivity), and one of 48 t1 cases was MM+ (98% specificity). DDC levels in peritoneal washes from patients with synchronous peritoneal metastases were more than 50 times higher than in those from patients without metastasis (P<0.01). For 15 cases of peritoneal dissemination (seven cases were cytologically positive), DDC was positive in 13 cases (87% sensitivity), but CEA failed to detect micrometastases in four cases (73% sensitivity), indicating that DDC is in some cases superior to CEA for the detection of peritoneal micrometastases of gastric cancer in terms of sensitivity as well as specificity, especially for poorly differentiated adenocarcinomas. A combination of CEA and DDC improved the accuracy of diagnosis up to 94%. These results suggest that DDC is potentially a novel marker for peritoneal dissemination of gastric cancer and that quantitative RT-PCR of DDC is reliable and efficient for the selection of patients for adjuvant intraperitoneal chemotherapy to prevent peritoneal recurrence. PMID- 14760383 TI - Fhit expression in human gastric adenomas and intramucosal carcinomas: correlation with Mlh1 expression and gastric phenotype. AB - The fragile histidine triad (FHIT) gene, encompassing the FRA3B fragile site at chromosome 3p14.2, is a candidate tumour suppressor gene involved in a variety of tumours, including gastric carcinomas. Recently, it has been reported that the FHIT gene may be a target of damage in some of mismatch-deficient tumours. To clarify further the role of the Fhit protein in gastric carcinogenesis, we investigated whether Fhit expression in early gastric neoplasia is associated with mismatch repair protein expression and cellular phenotype. Fhit, Mlh1 and phenotypic expression were evaluated immunohistochemically in 87 early gastric neoplasias, comprising 32 adenomas and 55 intramucosal carcinomas, resected by endoscopic mucosal resection therapy. Significant loss or reduction of Fhit expression was noted in four (12.5%) of the 32 adenomas and 21 (38.2%) of the 55 intramucosal carcinomas. The rate of abnormal Fhit expression was significantly higher in intramucosal carcinomas than in adenomas (P=0.021). Moreover, reduced Fhit expression was found to be significantly associated with loss of Mlh1 expression in early gastric neoplasia (P=0.0011). Furthermore, we also detected a significant association between reduced Fhit expression and gastric phenotype (P=0.0018). These results suggested that reduced Fhit expression occurs in the early stage of gastric carcinogenesis and could be correlated with a lack of Mlh1 expression and gastric phenotype. PMID- 14760384 TI - TP53 mutations in early-stage ovarian carcinoma, relation to long-term survival. AB - We conducted the present study to evaluate the frequency and prognostic importance on long-term survival of TP53 mutations and TP53 protein accumulation in a cohort of 178 patients with early-stage ovarian carcinomas. TP53 mutations scored as aberrant temporal temperature gradient gel electrophoresis pattern from all exons were observed in 39.9% of the tumours. Full screening of exons 5-8, followed by sequencing, was successful in 135 cases, and 48 mutations altering the protein were detected in 39 cases (28.9%). TP53 mutations were slightly less common in the Federation of Gynecologists and Obstetricians stage IA than in IB/IC (P=0.05). No significant correlations with histological type, grade of differentiation, DNA ploidy status or age at diagnosis were found. TP53 protein accumulation analysed by immunohistochemistry was found in 32.6% of all tumours, and was a poor predictor of TP53 mutations with 56.4% sensitivity, 77.1% specificity, 50% positive predictive value and 81.3% negative predictive value. Neither TP53 mutations nor TP53 protein accumulation influenced the prognosis significantly in this group of patients. PMID- 14760385 TI - Predicting biomarkers for ovarian cancer using gene-expression microarrays. AB - Ovarian cancer has the highest mortality rate of gynaecological cancers. This is partly due to the lack of effective screening markers. Here, we used oligonucleotide microarrays complementary to approximately 12 000 genes to establish a gene-expression microarray (GEM) profile for normal ovarian tissue, as compared to stage III ovarian serous adenocarcinoma and omental metastases from the same individuals. We found that the GEM profiles of the primary and secondary tumours from the same individuals were essentially alike, reflecting the fact that these tumours had already metastasised and acquired the metastatic phenotype. We have identified a novel biomarker, mammaglobin-2 (MGB2), which is highly expressed specific to ovarian cancer. MGB2, in combination with other putative markers identified here, could have the potential for screening. PMID- 14760386 TI - Lymphatic density and metastatic spread in human malignant melanoma. AB - Malignant melanoma (MM), the most common cause of skin cancer deaths, metastasises to regional lymph nodes. In animal models of other cancers, lymphatic growth is associated with metastasis. To assess if lymphatic density (LD) was increased in human MM, and its association with metastasis, we measured LD inside and around archival MM samples (MM, n=21), and compared them with normal dermis (n=11), basal cell carcinoma (BCC, n=6) and Merkel cell carcinoma (MCC), a skin tumour thought to metastasise through a vascular route (MCC, n=6). Lymphatic capillary density (mm(-2)), as determined by immunohistochemical staining with the lymphatic specific marker LYVE-1, was significantly increased around MM (10.0+/-2.5 mm(-2)) compared with normal dermis (2.4+/-0.9 mm(-2)), BCC (3.0+/-0.9 mm(-2)) and MCC (2.4+/-1.4 mm(-2)) (P<0.0001). There was a small decrease in LD inside MM (1.1+/-0.7 mm(-2)) compared with normal dermis, but a highly significant decrease in BCC (0.14+/-0.13) and MCC (0.12+/-2.4) (P<0.01 Kruskal-Wallis). Astonishingly, LD discriminated between melanomas that subsequently metastasised (12.8+/-1.6 mm(-2)) and those that did not (5.4+/-1.1 mm(-2), P<0.01, Mann-Whitney). Lymphatic invasion by tumour cells was seen mainly in MM that metastasised (70% compared with 12% not metastasising, P<0.05 Fisher's Exact test). The results show that LD was increased around MMs, and that LD and tumour cell invasion of lymphatics may help to predict metastasis. To this end, a prognostic index was calculated using LD, lymphatic invasion and thickness that clearly discriminated metastatic from nonmetastatic tumours. PMID- 14760387 TI - Simultaneous expression of COX-2 and mPGES-1 in mouse gastrointestinal hamartomas. AB - Cyclo-oxygenase (COX)-2 is induced in various types of cancer tissues. Here, we demonstrate stromal expression of both COX-2 and microsomal prostaglandin E(2) synthase (mPGES)-1 in gastrointestinal hamartomas developed in Lkb1(+/-), Smad4(+/-) and Cdx2(+/-)mice. These results suggest that PGE(2) produced by COX-2 and mPGES-1 plays an important role in hamartoma development regardless of the mutated genes causing hamartomas. PMID- 14760388 TI - ZD6126 inhibits orthotopic growth and peritoneal carcinomatosis in a mouse model of human gastric cancer. AB - The purpose of this study was to examine the effects of ZD6126, a novel vascular targeting agent, on tumour growth and angiogenesis in an orthotopic model of gastric cancer. TMK-1 human gastric adenocarcinoma cells were injected into the gastric wall of nude mice. After the tumours were established (day 14), therapy was initiated. Mice (n=11-12/group) received (a). vehicle, (b). ZD6126 at 100 mg x kg day(-1) i.p. one time per week or (c) ZD6126 at 100 mg x kg day(-1) i.p. five times per week. Tumour mass, volume and the presence or absence of peritoneal carcinomatosis were determined at sacrifice on day 38. Tumours from each group were stained for markers of blood vessels, proliferation and apoptosis. To further define the time frame of the vascular-targeting effects of chronic therapy with ZD6126, TMK-1 cells were again injected into the gastric wall of mice in a second experiment. On day 14, a single i.p. injection of ZD6126 100 mg x kg(-1) mouse(-1) or vehicle was delivered. Groups of three mice each were killed and the tumours harvested at days 1, 3 and 5 post-ZD6126 injection. Tumours were processed and stained for endothelial and tumour cell apoptosis and proliferation. No overt toxicity was observed with ZD6126 therapy. ZD6126 led to a marked inhibition of tumour growth (82% decrease vs control (P<0.001)). ZD6126 also led to a significant decrease in the incidence of peritoneal carcinomatosis (10 out of 12 controls, vs one out of 12 ZD6126) (P<0.01). Histological analysis of tumours revealed large regions of central necrosis in the treated group, as well as a dramatic increase in tumour cell apoptosis (7.4-fold increase (P<0.001)), consistent with the vascular-targeting activity of ZD6126. Mice treated with ZD6126 demonstrated a 59% decrease in PCNA-positive cells (P< 0.02), indicating reduced tumour cell proliferation. In addition, tumours treated with ZD6126 exhibited a 40% decrease in microvessel density (P<0.05). Results from mice treated with a single injection of ZD6126 demonstrated the acute effects this agent has on the tumour vasculature. The ratio of endothelial cell apoptosis to endothelial cell proliferation was increased within 24 h of a single injection. In conclusion, ZD6126 significantly inhibited tumour growth and metastasis in an orthotopic model of human gastric adenocarcinoma, without detectable problematic adverse effects. These data suggest that ZD6126 may be worthy of investigation in the treatment of primary gastric adenocarcinoma. PMID- 14760389 TI - Cyclooxygenase-2 selective inhibition with NS-398 suppresses proliferation and invasiveness and delays liver metastasis in colorectal cancer. AB - Nonsteroidal anti-inflammatory drugs (NSAIDs) have been reported to reduce the risk and mortality of colorectal cancer (CRC) by inhibiting the activity of cyclooxygenase (COX). The present studies were directed to determine whether selective COX-2 inhibition reduces CRC tumour cell proliferation and invasion/migration, and the possible cellular and molecular mechanisms involved. The MC-26 cells are a highly invasive mouse CRC cell line expressing COX-2 protein. NS-398 (100 microM), a highly selective COX-2 inhibitor, decreased cell proliferation by approximately 35% of control, as determined using [(3)H] thymidine incorporation. This reduction in cell proliferation was associated with decreased expression of cyclin D1 and proliferating cell nuclear antigen (PCNA). Furthermore, NS-398 inhibited cell invasion/migration through Matrigel extracellular matrix components at 24 h by approximately 60%. The addition of exogenous prostaglandin E(2) partially attenuated the inhibition of cell invasion by 10 microM NS-398, but failed to reverse the effect of 100 microM NS-398. Matrix metalloproteinases-2 (MMP-2) and -9 (MMP-9) are two enzymes that facilitate cell invasion/migration by degrading the extracellular matrix. In the presence of 100 microM NS-398, Western blot hybridisation analysis and zymography demonstrated that both MMP-2 and MMP-9 protein levels and enzyme activity were decreased by approximately 25-30%. In separate studies, NS-398 also inhibited tumour growth in vivo and retarded the formation of liver metastasis. The results of these studies indicate that the expression and activity of COX-2 appear to be associated with both the proliferative and invasive properties of CRC. Cyclooxygenase-2 inhibition suppresses tumour cell growth and invasion/migration, and retards liver metastasis in a mouse colon cancer model, via multiple cellular and molecular mechanisms. PMID- 14760390 TI - Mucinous differentiation features associated with hormonal escape in a human prostate cancer xenograft. AB - Many theories mention hypersensitive, promiscuous, outlaw or bypass signalling pathways to explain the acquisition of hormone independence in prostate cancer. Hormonal escape of prostate tumours is marked by many biological changes, including mucinous and neuroendocrine differentiation. Since expression of several mucins has been linked to carcinoma tumour progression, we have characterised the expression of mucins at both RNA and protein levels in an in vivo model of prostate cancer in hormonal escape. Using PAC120, a xenograft of a human hormone-dependent prostate tumour, and its hormone-independent variants, we analysed the expression of mucins (MUC1, MUC2, MUC4, MUC5AC, MUC5B, MUC6) by immunohistochemistry or reverse transcriptase (RT)-PCR. While the parental PAC120 tumour was a compact poorly-differentiated tumour of Gleason score 9 (5+4), hormone-independent variants displayed mucinous, neuroendocrine-like or mixed histological changes; these changes were stable through serial transplantations or after testosterone supply. MUC1 mRNA was expressed in both PAC120 and the hormone-independent variants, although at variable levels. All tumours displayed a high and constant expression of MUC2 and no expression of MUC4 mRNA. While MUC1 was expressed in all xenografts whatever their hormone dependence status, MUC2, MUC5B and MUC6 were preferentially expressed in hormone-independent variants. The loss of hormone dependence in this prostate cancer xenograft model is therefore marked by irreversible histological alterations, mucinous or neuro-endocrine, associated with an expression of secretory MUC2, MUC5B and MUC6, independent of the histological differentiation subtype. These data point to mucinous differentiation as an important step in the acquisition of hormone independence in this cancer, and suggest that secretory mucins might participate in an unknown pathway of hormonal escape in prostate cancer. PMID- 14760391 TI - Evidence that involucrin, a marker for differentiation, is oxygen regulated in human squamous cell carcinomas. AB - The majority of hypoxic cells in squamous cell carcinomas of the head and neck and cervix express involucrin, a molecular marker for differentiation. This raises the question of whether involucrin is an oxygen-regulated protein and, if so, whether it could serve as an endogenous marker for tumour hypoxia. Consistent with oxygen regulation, involucrin protein was found to increase with increasing hypoxia in confluent cultures of moderately differentiated human SCC9 cells. Cells harvested at the point of confluence and exposed to graded concentrations of oxygen revealed a K(m) of approximately 15 mmHg for involucrin induction. This is similar to K(m)s for HIF-1alpha, CAIX and VEGF. Involucrin induction showed a steep dependence on pO(2) with a transition from minimum to maximum expression occurring over less than an order of magnitude change in pO(2). In contrast to SCC9 cells, involucrin was not induced by hypoxia in poorly differentiated SCC4 cells. It is concluded that involucrin is an oxygen-regulated protein, but that differentiation modulates its transcription status with respect to hypoxia induction. PMID- 14760392 TI - Pharmacokinetics in mice and growth-inhibitory properties of the putative cancer chemopreventive agent resveratrol and the synthetic analogue trans 3,4,5,4' tetramethoxystilbene. AB - Resveratrol (trans-3,5,4'-trihydroxystilbene) is a naturally occurring polyphenol with cancer chemopreventive properties in preclinical models of carcinogenesis, including those of colorectal cancer. Recently, a variety of analogues of resveratrol have been synthesised and investigated in in vitro assays. One analogue, 3,4,5,4'-tetramethoxystilbene (DMU 212), showed preferential growth inhibitory and proapoptotic properties in transformed cells, when compared with their untransformed counterparts. As part of a chemoprevention drug development programme, the pharmacokinetic properties of DMU 212 were compared with those of resveratrol in the plasma, liver, kidney, lung, heart, brain and small intestinal and colonic mucosa of mice. DMU 212 or resveratrol (240 mg kg(-1)) were administered intragastrically, and drug concentrations were measured by HPLC. Metabolites were characterised by cochromatography with authentic reference compounds and were identified by mass spectrometry. The ratios of area of plasma or tissue concentration vs time curves of resveratrol over DMU 212 (AUC(res)/AUC(DMU212)) for the plasma, liver, small intestinal and colonic mucosa were 3.5, 5, 0.1 and 0.15, respectively. Thus, resveratrol afforded significantly higher levels than DMU 212 in the plasma and liver, while DMU 212 exhibited superior availability compared to resveratrol in the small intestine and colon. Resveratrol was metabolised to its sulphate or glucuronate conjugates, while DMU 212 underwent metabolic hydroxylation or single and double O-demethylation. DMU 212 and resveratrol inhibited the growth of human-derived colon cancer cells HCA 7 and HT-29 in vitro with IC(50) values of between 6 and 26 microM. In the light of the superior levels achieved in the gastrointestinal tract after the administration of DMU 212, when compared to resveratrol, the results provide a good rationale to evaluate DMU 212 as a colorectal cancer chemopreventive agent. PMID- 14760393 TI - Oral tegafur/folinic acid chemotherapy decreases phenytoin efficacy. PMID- 14760394 TI - Reversal of sensorimotor gating deficits in Brattleboro rats by acute administration of clozapine and a neurotensin agonist, but not haloperidol: a potential predictive model for novel antipsychotic effects. AB - Prepulse inhibition (PPI) of acoustic startle is decreased in unmedicated schizophrenia patients and similar deficits can be induced in rats through pharmacological, environmental, or neuroanatomical manipulations. Recently, we reported that Brattleboro (BB) rats, a Long Evans (LE) strain with a single gene mutation, have inherent deficits in PPI homologous to those observed in schizophrenia patients. We also reported that PPI deficits in BB rats could be reversed by chronic but not acute administration of 0.5 mg/kg haloperidol. No other dose or drug was tested in that experiment. In this study, we tested the effects of acute subcutaneous administration of several doses of haloperidol as well as the second-generation antipsychotic, clozapine, and the putative novel antipsychotic, PD149163, a neurotensin mimetic that crosses the blood-brain barrier. Consistent with our previous report, BB rats exhibited PPI deficits compared to LE rats and none of the doses of haloperidol produced a significant effect on this PPI deficit. In contrast, 10 and 15 mg/kg of clozapine and all the doses of PD149163 tested reversed the PPI deficits in BB rats. In addition, haloperidol, but not clozapine or PD149163 produced significant catalepsy in BB rats, supporting the notion that PD149163 has a profile consistent with atypical antipsychotics and providing support for the predictive validity of the PPI results. These results further strengthen the notion that the BB rat is a useful predictive model of antipsychotic efficacy and suggest that this model may differentiate between antipsychotics belonging to different therapeutic categories, for example, first- and second-generation agents. PMID- 14760395 TI - Trading scientific freedom. PMID- 14760397 TI - US pressures publishers to honor trade embargoes. PMID- 14760399 TI - Iraq's public health infrastructure a casualty of war. PMID- 14760401 TI - World agencies try to stem flood of fake drugs. PMID- 14760402 TI - For HIV vaccine trials, size does matter. PMID- 14760403 TI - In world of epidemics, WHO's in control. PMID- 14760406 TI - A lab of her own. PMID- 14760404 TI - US soldiers refuse to fall in line with anthrax vaccination scheme. PMID- 14760407 TI - Jeff Friedman. PMID- 14760408 TI - Specificity of mechanisms for plaque removal after A beta immunotherapy for Alzheimer disease. PMID- 14760410 TI - International cooperation on xenotransplantation. PMID- 14760411 TI - The 'apparent' color of money. PMID- 14760414 TI - Sweet relief for Huntington disease. PMID- 14760415 TI - Lipid unites disparate syndromes of sepsis. PMID- 14760416 TI - Inflamed about obesity. PMID- 14760417 TI - EMSY, a BRCA-2 partner in crime. PMID- 14760418 TI - Cities spawn epidemic dengue viruses. PMID- 14760422 TI - gp41: HIV's shy protein. PMID- 14760419 TI - Ceramide lances the lungs. PMID- 14760423 TI - Genetic, cellular and immune approaches to disease therapy: past and future. AB - Advances in immunology and molecular genetics have accelerated our understanding of the genetic and cellular basis of many diseases. At the same time, remarkable progress in recombinant DNA technology has enabled the development of molecular and cellular treatments for infectious diseases, inherited disorders and cancer. This Perspective is intended to give a sample of the progress over the past ten years in cellular, genetic and immune therapy of disease. During this time, monoclonal antibody technology and cellular transplantation have begun to come of age in biomedicine. Innovations in gene delivery have not only catalyzed the nascent field of human gene therapy, but may also ultimately impact human health by advancing recombinant vaccine technology. PMID- 14760427 TI - Cytokines as therapeutic targets for the gastrointestinal manifestations of scleroderma. AB - Systemic sclerosis (SSc), or scleroderma, is a connective tissue disorder characterized by progressive fibrosis of the skin and internal organs. It has significance for gastroenterologists because the gastrointestinal tract is involved in 90% of SSc patients, who often present with esophageal dysfunction. Though the exact pathogenesis of SSc is unknown, there is increasing evidence supporting an immune mechanism. Cytokines are the soluble mediators of immune activation, altered fibroblast proliferation and extracellular matrix accumulation in SSc and thereby provide important therapeutic targets. In the present review, the involvement of cytokines in SSc is discussed with particular emphasis on cytokines and growth factors that have been implicated in the disease process and likely play an important role in the gastrointestinal manifestations of scleroderma. The role of cytokines as therapeutic targets in scleroderma forms the basis of this timely review. PMID- 14760428 TI - Strongly positive tissue transglutaminase antibody assays without celiac disease. AB - Celiac disease is a small bowel disorder characterized by flattened villi and crypt hyperplasia, often with malabsorption. Improvement occurs with a gluten free diet. Sensitive and specific assays (eg, immunoglobulin A antibodies to tissue transglutaminase [tTG]) that can be quantified appear to be valuable tools for population screening studies. In addition, their use is expanding widely in the clinical practice arena, being employed as a method of case finding. In this evaluation, clinical use of a commercially available test kit was explored. Of 1330 samples submitted to our hospital laboratory by physicians in British Columbia, Alberta and the Yukon Territory (from 1999 to 2003, inclusive), 96 patients (7%) had increased values (normal range greater than 20 units) and markedly increased levels greater than 100 units were detected in 36 patients (3%). Of these, 14 patients (almost 40%) were referred to gastroenterologists in our hospital and all 14 had small intestinal biopsies. Of these, three patients (more than 20%) did not have celiac disease. Two had normal small bowel biopsies and one had unclassified sprue or 'sprue-like' intestinal disease that failed to respond to a gluten-free diet. The other 11 had biopsy-defined celiac disease. While the tTG assay may be a useful predictor of celiac disease, small intestinal biopsy is still required to confirm the diagnosis. In clinical practice, even strongly positive tTG results are not specific in individual patients, do not necessarily correlate with the degree of severity of biopsy change and, as a result, are also unlikely to be useful for monitoring diet compliance. PMID- 14760429 TI - Birth of the Canadian Digestive Health Foundation. AB - The Canadian Digestive Disease Foundation, renamed the Canadian Digestive Health Foundation--Fondation canadienne pour la promotion de la sante digestive--in December 2001, is the culmination of ongoing efforts by the Canadian Association of Gastroenterology to establish an independent charitable organization. In February 2001, it was officially endorsed as the Foundation for the Canadian Association of Gastroenterology. The initial efforts to establish this Foundation, led by Dr Richard McKenna in 1963, were unsuccessful. In 1991, Glaxo Canada (now GlaxoSmithKline) became a founding donor, and with the four founding physicians--Drs Ivan T Beck, Richard H Hunt, Suzanne E Lemire and Alan BR Thomson -the expenses to establish the Foundation were met. A charitable number was obtained in 1995 (0997427-11). The second founding donor was Janssen Canada (now Janssen-Ortho), and public education support came from Astra Canada (now AstraZeneca Canada). The Foundation initially relied on corporate donors, but now approaches physicians, patients and the general public. The objectives of the Foundation are to advance the science of gastroenterology and to provide knowledge of digestive diseases and nutrition to the general public, to enhance the quality of life of persons who are afflicted with these disorders. The major achievements of the Foundation are the provision of one-year operating grants to new investigators, which have allowed them to accumulate early data and subsequently obtain support from other major granting organizations. It also provides Fellowships and studentship support grants, in conjunction with the Canadian Institutes of Health Research and the pharmaceutical industry. The education committee found that there was little research support in this field, considering the large economic burden of digestive disease and the amount of outstanding work done by Canadian researchers. A bilingual Web site, a web-based specialist's discussion program and bilingual pamphlets facilitate public awareness and allow patients to voice concerns. PMID- 14760430 TI - Functional development of the human gastrointestinal tract: hormone- and growth factor-mediated regulatory mechanisms. AB - The present review focuses on the control of gastrointestinal (GI) tract development. The first section addresses the differences in general mechanisms of GI development in humans versus rodents, highlighting that morphogenesis of specific digestive organs and the differentiation of digestive epithelia occur not only at different stages of ontogeny but also at different rates. The second section provides an overview of studies from the author's laboratory at the Universite de Sherbrooke pertaining to the development of the human fetal small intestine and colon. While both segments share similar morphological and functional characteristics, they are nevertheless modulated by distinct regulatory mechanisms. Using the organ culture approach, the author and colleagues were able to establish that hormones and growth factors, such as glucocorticoids, epidermal growth factor, insulin and keratinocyte growth factor, not only exert differential effects within these two segments, they can also trigger opposite responses in comparison with animal models. In the third section, emphasis is placed on the functional development of human fetal stomach and its various epithelial cell types; in particular, the glandular chief cells responsible for the synthesis and secretion of gastric enzymes such as pepsinogen 5 and gastric lipase. Bearing in mind that limitations of available cell models have, until now, greatly impeded the comprehension of molecular mechanisms regulating human gastric epithelial cell functions, the last section focuses on new human gastric epithelial cell models recently developed in the author's laboratory. These models comprise a novel primary culture system of human fetal gastric epithelium including, for the first time, functional chief cells, and human gastric epithelium cell lines cloned from the parental NCI-N87 strain. These new cells lines could serve important applications in the study of pathogenic action and epithelial regeneration. PMID- 14760431 TI - Complementary and alternative medicine use by patients with inflammatory bowel disease: are Canadian physicians failing with conventional therapy, or not? PMID- 14760433 TI - The hereditary spectrum of pancreatic cancer: the Edmonton experience. AB - OBJECTIVE: Pancreatic cancer is known to aggregate in some families and has been associated with a wide variety of cancer syndromes. The authors describe their experience with pancreatic cancer and the range of associated cancer syndromes. METHODS: The charts of all patients seen for concern of a hereditary cancer syndrome in the Cancer Genetics Clinic at the University of Alberta between 1995 and 2002 were reviewed. RESULTS: Forty families reported a personal or family history of pancreatic cancer in the context of a possible hereditary cancer syndrome. Three additional families reported a history of pancreatitis. Twenty four (56%) of those families were suspected of having a hereditary breast and ovarian cancer syndrome. A further seven (16%) were suspected of having hereditary nonpolyposis colon cancer. Only three (7%) were believed to be at risk for a site-specific pancreatic cancer syndrome. Another three (7%) were suspicious for hereditary pancreatitis. The remaining family histories were suggestive of Li-Fraumeni syndrome, von Hippel-Lindau syndrome or a nonspecific cancer predisposition. CONCLUSIONS: With such a wide variety of hereditary cancer syndromes associated with pancreatic cancer, an accurate assessment of the family history is essential to determine the most appropriate cancer screening for at risk family members and to guide any molecular testing that may be offered. PMID- 14760434 TI - Thanks, Toronto. PMID- 14760435 TI - To rationalize rationing. PMID- 14760437 TI - Lack of association between activity or fitness and loss of control of hypertension. AB - BACKGROUND: Many treated hypertensive patients have inadequate control of their blood pressure. Physical activity and cardiorespiratory fitness are important in the development and treatment of hypertension. Less is known about their relationships with hypertension control in the context of long-term management. OBJECTIVES: To identify whether patients with lower levels of activity or fitness are at risk for loss of control of hypertension. METHODS: Follow-up blood pressure measurements were recorded over the period of a year in a cohort of initially controlled hypertensive patients, under the care of 50 family physicians in a variety of practice settings. A questionnaire for a one-year recall of leisure time physical activity and the Step Test, an indirect measure of maximal oxygen consumption, were administered to the patients at the end of the period of blood pressure observations. The study group for the activity questionnaire consisted of 385 patients; 310 of whom undertook the fitness test. Sociodemographic, clinical and lifestyle factors were analyzed as potential confounders in the relationships of main interest in this study: between activity and fitness, respectively, with loss of control of hypertension. RESULTS: Sixteen per cent of patients (95% CI 12, 20) had an increase of 20 mmHg systolic or 10 mmHg diastolic from baseline to the average of three follow-up blood pressures, and 14% (95% CI 11, 18) had blood pressures above the entry thresholds on at least two of three follow-up visits. No associations were found between loss of control of hypertension and physical activity or cardiorespiratory fitness. CONCLUSIONS: Lower levels of physical activity and fitness are not associated with loss of hypertension control. The identification of patients at risk for loss of hypertension control, which could be useful in targeting selected groups for increased follow-up and in understanding the high prevalence of uncontrolled hypertension, requires further study. PMID- 14760438 TI - The number needed to harm: is it too optimistic? AB - The number needed to treat (NNT) to prevent one adverse outcome event is a useful measure of clinical trial results and represents the therapeutic effort required to realize a specific clinical benefit. With many therapies, there are both positive and negative consequences. Analogous to the NNT, there is the number needed to harm (NNH), which is the reciprocal of the absolute risk increase in situations where the experimental treatment harms more patients then the control treatment. The NNH is the number of patients who, if they received the experimental treatment, would lead to one additional person being harmed compared with patients who receive the control treatment. The NNH may, however, provide an optimistic measure of the true risks caused by therapy. PMID- 14760439 TI - Transcatheter management of residual shunts after initial transcatheter closure of a patent arterial duct. AB - OBJECTIVES: To assess the efficacy and safety of transcatheter reocclusion of persistent leaks following previously attempted transcatheter occlusion of persistent arterial duct. DESIGN: Retrospective study. SETTING: Tertiary pediatric cardiology centre. PATIENTS: From February 1987 through October 1996, trans-catheter occlusion of a residual ductal shunt was attempted in 42 consecutive patients at a median age of 5.0 years (range 1.6 years to 16.2 years). INTERVENTIONS: Fourty patients had successful placement of a double umbrella occluder (n=27) or coils (n=13) across residual shunts. Complications included device embolization in two patients and hemolysis in one patient. OUTCOME MEASURES AND RESULTS: Mean z-score for left ventricular end-diastolic dimension (LVEDD) at initial echocardiography was +2.55 +/- 1.89 (P<0.0001 versus normal); z-score for left pulmonary artery (LPA) diameter was +2.00 +/- 1.52 (P<0.0001). Mean LPA to right pulmonary artery (RPA) diameter ratio was 1.05 +/- 0.18. At follow-up echocardiogram, a median of two years (range six months to 7.7 years) after the second procedure, a shunt was persistent in 3% of the patients. Mean LVEDD and LPA diameter z-value, and mean LPA to RPA diameter had dropped significantly to +0.42 +/- 1.31, +0.07 +/- 1.15 and 0.86 +/- 0.14 (P<0.001), respectively. LPA flow acceleration was present in 25% of patients. Three of nine patients, in whom lung perfusion scan was performed, had left lung perfusion below 40%. Small weight and age at catheterization were significant risk factors for LPA flow disturbance. CONCLUSIONS: Repeat transcatheter occlusion is safe and successful in eliminating residual shunt across the arterial duct. Attention should, however, be addressed to the potential for LPA stenosis and growth, and flow should be regularly assessed. PMID- 14760440 TI - Management of dyslipidemia in primary care. AB - BACKGROUND: Cardiovascular disease remains the leading cause of mortality in Canada. The link between hyperlipidemia and coronary heart disease has been clearly established. There is overwhelming evidence for reductions in coronary events and cardiovascular mortality with lowering of low-density lipoprotein cholesterol (LDL-C). Despite the evidence, hyperlipidemia treatment remains suboptimal. OBJECTIVE: To evaluate compliance with published dyslipidemia guidelines in a primary care setting. The primary outcome measure was target LDL C level. METHODS: Retrospective chart review of a random selection of 300 patients diagnosed with hyperlipidemia in a large academic family medicine clinic. The primary outcome measure was a target LDL-C level of less than 2.5 mmol/L for patients with diabetes or coronary heart disease. For patients without diabetes or coronary heart disease, Framingham risk assessment tables were used to determine ideal target LDL-C levels. RESULTS: Overall, 53% of patients achieved target LDL-C. Target LDL-C levels were achieved in 48% of patients with diabetes or coronary heart disease. Males were twice as likely to be prescribed lipid lowering therapy than females. Males on lipid lowering therapy were twice as likely as females on lipid lowering therapy to achieve target LDL-C levels. Males with diabetes or coronary heart disease were twice as likely as females with diabetes or coronary heart disease to achieve target LDL-C levels. Only 44% of patients with diabetes or coronary heart disease were prescribed lipid lowering therapy. CONCLUSION: Results from an academic family medicine clinic indicate suboptimal compliance with current dyslipidemia management guidelines. Primary care physicians need to continue to take an aggressive stance on lipid lowering strategies, especially in high-risk patients and females. PMID- 14760441 TI - Predictors of early reangiography within 30 days after coronary stenting. AB - BACKGROUND: A substantial number of early urgent reangiographies after coronary stenting do not reveal coronary abnormalities such as thrombosis, dissection, restenosis or side branch occlusion. The characterization of patients undergoing early reangiography may reduce the number of potentially unnecessary procedures. OBJECTIVE: To evaluate the predictors for unplanned early re- angiography on the basis of the information available at the time of stent implantation. METHODS: All 71 patients with reangiography after stent implantation between 1994 and 1998 in the Department of Cardiology at the University Hospital of Vienna, Austria, were compared with a control sample of 88 patients without early reangiography during the same period (control subjects were matched for the time point of the first intervention). The clinical and procedural variables were analyzed in this case-control study. For specification of the group with negative reangiograms, differences in clinical parameters between patients with negative versus patients with positive reangiograms were also analyzed. RESULTS: Clinical predictors for early reangiography of patients without evidence of important coronary pathology were a history of hypertension and left ventricular hypertrophy, as well as total cholesterol. The angiographic predictor was multiple vessel disease, and procedural risk factors included the left anterior descending artery as the target artery, type B and C lesions and total occlusion in the left anterior descending artery, as well as high-pressure balloon stent deployment. CONCLUSION: Clinical, angiographic and procedural variables predict the risk for unplanned early reangiography after coronary stenting. Hypertensive heart disease may mimic acute coronary events; hypertension and left ventricular hypertrophy represent independent predictors of unplanned reangiography in patients with good short term results. PMID- 14760442 TI - Using left atrial esophagus impression after a barium swallow to determine the optimal septal puncture site during percutaneous balloon mitral valvuloplasty. AB - BACKGROUND: Transseptal catheterization is the key to a successful percutaneous balloon mitral valvuloplasty (PBMV). The primary aim of the study was to investigate new methods of transseptal catheterization for PBMV. METHODS: Eighty four patients with rheumatic mitral stenosis were randomized into two groups for PBMV. In the study group, the left atrial (LA) impression on the esophagus after a barium swallow was used as a reference of LA silhouette to determine the septal puncture site. In the control group, the LA silhouette under a normal fluoroscopic view was used for the determination of septal puncture site. RESULTS: In the study group, the average length of esophagus impression after a barium swallow was similar to the size of the left atrium measured by angiography (43.8+/-0.6 mm versus 44.1+/-0.7 mm, n=42, P>0.05). The success rate of atrial septum puncture in the study and control groups were 100% and 64.3%, respectively (P<0.01). Transseptal catheterization was subsequently achieved using the barium swallow approach in the 15 initially unsuccessful patients from the control group. PBMV was successful in the 84 patients with significant reduction in LA pressure and improvement of clinical symptoms. CONCLUSIONS: Transseptal catheterization in patients undergoing PBMV can be safely and effectively performed using the barium swallow approach. PMID- 14760443 TI - Na+/H+ exchange inhibition with HOE 642 improves recovery of the injured neonatal rabbit heart. AB - BACKGROUND: The effects of inhibition of the Na+/H+ exchanger (NHE) on postischemic recovery of the injured neonatal rabbit heart were examined. The NHE may be an important mechanism for reperfusion injury in the neonate heart. The effects of two NHE inhibitors, HOE 642 (HOE) and 5-(N,N-dimethyl)-amiloride (DMA), given during hypothermic cardioplegic arrest, were evaluated. METHODS: Isolated working crystalloid-perfused neonatal rabbit hearts were subjected to 10 min of normothermic ischemia to cause injury before undergoing 4 h of hypothermic (10 degrees C) cardioplegic arrest with a single dose of crystalloid solution (controls, n=21) or with the addition of 0.5 micromol/L HOE (n=24) or 30 micromol/L DMA (n=15). RESULTS: Hearts subjected to HOE had improved recoveries of aortic flow when compared with controls at 15 min and 30 min of reperfusion (35.7+/-1.3 mL/min versus 26.2+/-1.4 mL/min, respectively, at 15 min, P<0.0001; 36.5+/-1.5 mL/min versus 23.6+/-1.6 mL/min, respectively, at 30 min, P<0.0001) and with DMA at 30 min (36.5+/-1.5 mL/min versus 29.9+/-1.9 mL/min, P=0.0214). Cardiac output and systolic pressure were also improved at 30 min in HOE hearts versus controls (P<0.0001). CONCLUSIONS: NHE inhibition with HOE during cardioplegic arrest resulted in improved functional recovery of injured hearts. Further studies in blood-perfused neonatal preparations are warranted. PMID- 14760444 TI - Potential role of lysophosphatidic acid in hypertension and atherosclerosis. AB - BACKGROUND: Lysophosphatidic acid (LPA) is present in both serum and cytosol. Serum LPA is mainly released from platelets whereas cytosolic LPA is the metabolite of phosphatidic acid due to the action of phopholipase A2. Because platelet function and phospholipase A2 activity are upregulated in hypertensive and coronary heart disease patients, respectively, plasma and cytosolic LPA levels are expected to be higher in these pathological conditions. OBSERVATIONS: LPA is known to cause platelet aggregation and thus release more LPA as well as platelet-derived growth factor; this positive feedback circuit leads to the continuous growth of vascular smooth muscle cells (VSMCs). LPA also increases the intracellular concentration of free calcium in VSMCs and elevates the blood pressure. LPA content in the atherosclerotic plaque is elevated about 13 times in comparison with normal tissues because oxidized low-density lipoproteins promote the production of LPA. On the other hand, LPA has been shown to protect the heart from ischemia and reperfusion-induced damage due to its antiapoptosis effect. Because LPA has been reported to stimulate mitogen-activated protein kinase, phosphatidylinositide-3-kinase and protein kinase C, this bioactive phospholipid may be involved in the signal transduction mechanisms during the process of cardiac hypertrophy. CONCLUSIONS: Due to its ability to increase intracellular Ca2+ and proliferation of VSMCs, LPA may play an important role in the development of hypertension and atherosclerosis. It is therefore suggested that LPA antagonists may prove useful in the treatment of both hypertension and atherosclerosis. PMID- 14760445 TI - Folie a deux: an old diagnosis with a new technology. AB - The implantable cardioverter defibrillator (ICD) is the treatment of choice for life-threatening ventricular tachyarrhythmias. Despite this, indications for explantation have been described. Psychiatric complications have been considered in the decision to not re-implant a previously explanted device but never as a primary indication for explantation. The case of a woman who requested ICD explantation amidst concerns that the titanium used in its construction was absorbing harmful negative electromagnetic field energy from the environment resulting in intermittent bouts of fatigue, diaphoresis and an overall sense of impending doom is presented. She and her husband had significant psychosocial impairment because of this belief system. The present case is the first known report of a psychiatric complication as the primary indication for ICD explantation. It is illustrative of diagnostic dilemmas, competency and the importance of quality of life. PMID- 14760447 TI - An incidental calcified right atrial mass. AB - A 58-year-old man presented to his family physician with a mild cough. A routine chest x-ray revealed a 4 cm 'ring opacity' in the right atrium. A transthoracic echocardiogram revealed a 41 mm x 33 mm mass in the right atrium. The patient underwent urgent cardiac surgery, where a firm to hard 4 cm x 3 cm mass was excised. Pathology revealed an old, largely infarcted and calcified right atrial myxoma. A review of the published literature shows that this case is the first to demonstrate an almost completely infarcted atrial myxoma with only a few clusters of viable cells identified histologically. PMID- 14760451 TI - Water toxicity monitoring using Vibrio fischeri: a method free of interferences from colour and turbidity. AB - In this paper the kinetic method for the determination of toxicity using Vibrio fischeri is described and suggested as a potential method for the continuous screening of wastewater toxicity. The kinetic method was demonstrated to be free from interferences due to colour and turbidity normally observed when testing wastewater samples with this organism. This is of great importance for the application of the method to remote toxicity screening of wastewaters. The effect of colour, investigated using 50 ppm Zn(2+) solutions containing the food-dye tropaeolin O, and the effect of turbidity, investigated using 50 ppm Zn(2+) solutions containing white optically reflective and coloured optically absorbing polystyrene beads, is reported. It was also found that the design of the light detection system of the instrument ensures efficient collection of the light scattered by particles in the sample, which enables a greater range of turbid samples to be tested. In addition the natural light decay was found to be negligible during the duration of a 10 min test and thus one channel would be enough to carry out the tests. This would mean halving the quantity of bacterial reagent used and reducing the cost of the tests. PMID- 14760452 TI - Standard use conditions of terrestrial gastropods in active biomonitoring of soil contamination. AB - The rehabilitation of contaminated sites is becoming a rising preoccupation which requires the knowledge of their past before realization of a suitable remediation. Physicochemical analysis must be realized jointly with the use of bioindicator organisms, which, owing to their bioaccumulation capacities, will reveal the bioavailability of metals in soils. Among terrestrial invertebrates, gastropods like Helix aspersa aspersa possess an important organotropism for metals in their digestive gland and they can be used in active biomonitoring. During in situ monitoring, two parameters are tested: growth and accumulation of Fe, Mn, Zn, Cu, Cd, Ni, Pb, and Cr in viscera of snails. Environmental conditions, like humidity or autochthonous vegetation, are able to modify growth or bioaccumulation. In order to remove the variation of these factors, microcosms previously used must be improved: they were equipped with porous candles, which continually humidify soil. Concerning vegetation, an experimental plan was realized to determine the combination of food with the best compromise between growth and bioaccumulation: the combination clover-snail feed was chosen. Thus, in situ, the experimental environment will be repeated in all investigated sites and used to allow follow up of their contamination levels and intercomparison between different sites. PMID- 14760453 TI - Organotins in marine mammals and seabirds from Norwegian territory. AB - An increasing number of studies indicate that marine mammals and some seabirds are exposed to organotins. However, results from northern and Arctic areas are few. Here results from analysis of tributyltin (TBT), dibutyltin (DBT), monobutyltin (MBT), triphenyltin (TPhT), diphenyltin (DPhT) and monophenyltin (MPhT) in harbour porpoise (Phocoena phocoena), common seal (Phoca vitulina), ringed seal (Phoca hispida) and glaucous gull (Larus hyperboreus) from Norwegian territory are presented. Relatively high concentrations of DBT, TBT and MBT were observed in muscle, kidney and liver from harbour porpoises caught in northern Norway in 1988, just before restrictions on the use of tributyltin (TBT)(mainly on small boats) were introduced in several European countries. The concentrations in harbour porpoise muscle tissue were reduced significantly 11 years later, possibly as a result of the introduced restrictions. Considerably lower concentrations of butyltins were observed in the seals compared to porpoises. The lowest levels of organotins were found in ringed seals from Spitsbergen, where only traces of dibutyltin (DBT) and monobutyltin (MBT) were observed. Traces of DBT and MBT were also found in some individual glaucous gulls from Bear Island. The sum of the degradation products MBT and DBT in liver samples from all analysed species were generally higher than TBT itself. Triphenyltin (TPhT) was observed in all porpoise samples and in livers of common seals. Also the sum of the degradation products MPhT and DPhT in liver samples from porpoise and common seals were higher than TPhT. No traces of phenyltins were found in ringed seals from Spitsbergen or in glaucous gulls from Bear Island. The limited data available indicate low to moderate exposure to organotins in northern areas (Spitsbergen and Bear Island). Marine mammals are however more exposed further south along the Norwegian Coast. PMID- 14760454 TI - Viable fungi and bacteria in personal exposure samples in relation to microenvironments. AB - Personal exposures to viable fungi and bacteria were compared with the concentrations being assessed by stationary samplers in home and workplace microenvironments. A random sample of 81 elementary school teachers in eastern Finland performed two 24-hour measurement periods in wintertime. Concentrations and prevalences of viable fungi and bacteria on the collection filters were determined by cultivation method. The geometric mean concentration was 3-12 cfu m(-3) for total viable fungi, 0.6-3.7 cfu m(-3) for Penicillium and mainly under 1 cfu m(-3) for other fungi. The samples with higher fungal concentrations also had higher diversity of fungi than samples with lower concentrations. The total number of fungal genera recovered was 39 for personal, 34 for home and 23 for work samples. The variation in concentration of Penicillium explained even 25-95% of the variations of total fungal concentration in personal exposure, home and workplace environments. There was an association between personal exposure and home concentration of viable fungi and between personal exposure and home and work concentrations of viable bacteria. Personal exposure and home concentrations of fungi were higher in rural areas than in urban areas. Our results also indicate that presence of a certain fungus in a microenvironment does not necessarily mean similar findings in personal exposure samples. PMID- 14760455 TI - The application of bio-indicators for the assessment of air pollution. AB - Levels of metals were assessed in topsoil, dust, leaves, grass, lichen and bark from the Kruger National Park using electrothermal atomic absorption techniques. It was found that dust, lichen and bark reflected the levels of air pollution better than grasses or leaves of trees. It is suggested that the difference in levels of accumulated metals in the bark or lichens could be used as an indicator of air pollution in remote areas. PMID- 14760456 TI - Correlations in the chemical composition of rural background atmospheric aerosol in the UK determined in real time using time-of-flight mass spectrometry. AB - An aerosol time-of-flight mass spectrometer (ATOFMS) was used to determine, in real time, the size and chemical composition of individual particles in the atmosphere at the remote inland site of Eskdalemuir, Scotland. A total of 51,980 particles, in the size range 0.3-7.4 microm, were detected between the 25th and 30th June 2001. Rapid changes in the number density, size and chemical composition of the atmospheric aerosol were observed. These changes are attributed to two distinct types of air mass; a polluted air mass that had passed over the British mainland before reaching Eskdalemuir, interposed between two cleaner air masses that had arrived directly from the sea. Such changes in the background aerosol could clearly be very important to studies of urban aerosols and attempts at source apportionment. The results of an objective method of data analysis are presented. Correlations were sought between the occurrence of: lithium, potassium, rubidium, caesium, beryllium, strontium, barium, ammonium, amines, nitrate, nitrite, boron, mercury, sulfate, phosphate, fluorine, chlorine, bromine, iodine and carbon (both elemental and organic hydrocarbon) in both fine (d < 2.5 microm) and coarse (d > 2.5 microm) particle fractions. Several previously unreported correlations were observed, for instance between the elements lithium, beryllium and boron. The results suggest that about 2 in 3 of all fine particles (by number rather than by mass), and 1 in 2 of all coarse particles containing carbon, consisted of elemental carbon rather than organic hydrocarbon (although a bias in the sensitivity of the ATOFMS could have affected these numbers). The ratio of the number of coarse particles containing nitrate anions to the number of particles containing chloride anions exceeded unity when the air mass had travelled over the British mainland. The analysis also illustrates that an air mass of marine origin that had travelled slowly over agricultural land can accumulate amines and ammonium. PMID- 14760457 TI - Spatial and temporal variations of SPM, RPM, SO2 and NOx concentrations in an opencast coal mining area. AB - A study for assessment and management of air quality was carried out in the Ib Valley area of the Ib Valley coalfield in Orissa state, India. The 24 h average concentrations of suspended particulate matter (SPM), respirable particulate matter (RPM), sulfur dioxide (SO(2)) and oxides of nitrogen (NO(x)) were determined at regular intervals throughout one year at twelve monitoring stations in residential areas and six monitoring stations in mining/industrial areas. The 24 h average SPM and RPM concentrations were 124.6-390.3 microg m(-3) and 25.9 119.9 microg m(-3) in residential areas, and were 146.3-845.2 microg m(-3) and 45.5-290.5 microg m(-3) in industrial areas. During the study period, 24 h and annual average SPM and RPM concentrations exceeded the respective standards set in the Indian national ambient air quality standard (NAAQS) protocol as well as USEPA, EU, WHO and World Bank standards at most of the residential and industrial areas. However, concentrations of SO(2)(annual average: 24.6-36.1 microg m(-3) and 24 h average: 17.0-46.3 microg m(-3)) and NO(x)(annual average: 23.6-40.9 microg m(-3) and 24 h average: 18.3-53.6 microg m(-3)) were well within the prescribed limit of the NAAQS and international standards in both residential and industrial areas. The temporal variations of SPM and RPM fitted polynomial trends well and on average in the mining area 31.91% of the SPM was RPM. The linear regression correlation coefficients between SPM and RPM and between NO(x) and SO(2) were 0.94 (+/-0.04) and 0.66 (+/-0.10), respectively. The optimum interpolation technique, kriging, determined that maximal concentrations of SPM and RPM occurred within the mining site. Highest concentrations of particulate matter were observed during the winter season followed by summer, autumn and rainy seasons. An action plan is formulated for effective control of air pollution at source, and mitigative measures should include implementation of green belts around the sensitive areas where the concentration of air pollutants exceeds the standard limit. PMID- 14760458 TI - A combination of micro-porous membrane liquid-gas extraction and solid-phase trapping for ultra trace determination of benzene in urine. AB - A method was developed for the determination of benzene in urine. The sample was pumped through the donor channel of a membrane extraction unit with a micro porous membrane, separating the donor channel from an identical acceptor channel purged with nitrogen. The analyte reached the acceptor channel by diffusion through the membrane and was then swept by the carrier to a solid sorbent tube, where it was trapped. The analyte was subsequently thermally desorbed and analyzed by gas chromatography (GC) with mass selective detection (MS). After optimization, the recovery was close to quantitative, or 95%. Purging the membrane unit with pure water in between the samples eliminated any memory effects. The linearity was good in the concentration range examined (20-4000 ng l(-1)), with a correlation coefficient of 0.9996. The repeatability at 50 ng l( 1) and 400 ng l(-1) was 1.4% and 1.2%, respectively. The limit of detection was 12 ng l(-1) and the limit of quantification 35 ng l(-1). This enables assessment of benzene exposures of occupationally exposed subjects, of smokers and the majority of the general population. The developed method can be easily automated. PMID- 14760459 TI - Comparison of analytical results for chloride, sulfate and nitrate obtained from adjacent ice core samples by two ion chromatographic methods. AB - The anions chloride, sulfate and nitrate in nearly 500 pairs of ice core samples from the same depth were determined in a 121 m long ice core from Svalbard. The analyses were performed separately using an ion chromatograph with Dionex AS9 and AS15 columns with Na(2)CO(3) and NaOH eluents. Results showed a small (5-6 microg l(-1)) but statistically significant difference in mean concentrations for chloride and sulfate but not for nitrate. 2% of the data indicate real differences in concentrations across the ice core. Despite these differences ion information in ice core layers are comparable for ice core paleoclimate and environmental studies even though analyses are made using two different procedures. PMID- 14760461 TI - Evaluation of forest nutrition based on large-scale foliar surveys: are nutrition profiles the way of the future? AB - This paper introduces the use of nutrition profiles as a first step in the development of a concept that is suitable for evaluating forest nutrition on the basis of large-scale foliar surveys. Nutrition profiles of a tree or stand were defined as the nutrient status, which accounts for all element concentrations, contents and interactions between two or more elements. Therefore a nutrition profile overcomes the shortcomings associated with the commonly used concepts for evaluating forest nutrition. Nutrition profiles can be calculated by means of a neural network, i.e. a self-organizing map, and an agglomerative clustering algorithm with pruning. As an example, nutrition profiles were calculated to describe the temporal variation in the mineral composition of Scots pine and Norway spruce needles in Finland between 1987 and 2000. The temporal trends in the frequency distribution of the nutrition profiles of Scots pine indicated that, between 1987 and 2000, the N, S, P, K, Ca, Mg and Al decreased, whereas the needle mass (NM) increased or remained unchanged. As there were no temporal trends in the frequency distribution of the nutrition profiles of Norway spruce, the mineral composition of the needles of Norway spruce needles subsequently did not change. Interpretation of the (lack of) temporal trends was outside the scope of this example. However, nutrition profiles prove to be a new and better concept for the evaluation of the mineral composition of large-scale surveys only when a biological interpretation of the nutrition profiles can be provided. PMID- 14760460 TI - Mass loading of nickel and uranium on plant surfaces: application of laser ablation-ICP-MS. AB - Transport of contaminated sediments from a former radiological settling pond results in the deposition of U and Ni in the Lower Tims Branch (LTB)(Aiken, SC, USA). Uranium is unavailable for plant uptake, but elevated U and Ni concentrations associated with foliage of understory plants suggested mass loading. Mass loading of contaminated soil on Andropogon elliottii Chapman (Poaceae) was investigated using laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS). The technique allows for rapid quantitative elemental depth profiling. Fresh washed and unwashed leaves (n= 5) from the contaminated area were compared with those from an uncontaminated area, analysing Ni and U at ten randomly chosen points on each leaf. Nickel and U concentrations differed significantly between washed and unwashed leaves from LTB. Particles on unwashed leaves measured up to 300 [micro sign]m in diameter, and were enriched with U. Uranium was detected on the surface of the leaf, whereas Ni was detected within leaf tissues. In unwashed LTB leaves, Ni and U concentrations did not significantly differ in areas with and without visible particles, suggesting that there were much smaller particles, indistinguishable at [times]100 magnification, which contributed to the overall metal burden. Washing removed the majority of the Ni and U on the surface, but residual U and Ni was detected. Irregularities in the leaf surface, such as scars from herbivory contained elevated U concentrations despite a washing step, presumably from trapping soil particles. Laser ablation ICP-MS revealed that mass loading makes a significant contribution to the contaminant burden of understory plants at LTB. PMID- 14760463 TI - Laparoscopy for Crohn disease. AB - The laparoscopic approach to Crohn disease offers the multiple potential benefits of faster recovery, better cosmesis, and a lower incidence of adhesion-related complications and incisional hernias. Most of these advantages are unproven, but a review of the current literature reveals that advantages have been suggested in almost all series that have compared laparoscopy to laparotomy. Some features of Crohn disease, such as fistula, abscess, and phlegmon, may pose a surgical challenge; however, laparoscopic resection is often feasible, with morbidity rates comparable to or lower than those associated with laparotomy. Experience both in advanced laparoscopic techniques and surgery for Crohn disease, coupled with sound surgical judgment and a reasonably low threshold to convert to laparotomy before intraoperative complications occur, are essential for the successful and safe employment of these procedures. PMID- 14760464 TI - The use of laparoscopic techniques in surgery for mucosal ulcerative colitis. AB - The use of laparoscopic techniques when surgical therapy is required for the treatment of mucosal ulcerative colitis has been slow to develop; the surgery is extensive, and the instruments are limited. The often urgent nature of the surgery, along with the fragile inflamed colon, have contributed to the limited development of this surgical approach. Yet there is a paradox, as these patients have often anticipated surgery and thus are extremely frightened about it, or must undergo a major operation under urgent circumstances. In this setting, it would seem that a minimally invasive surgical approach would make sense if safe and reasonable, because healing, scarring, and patient fears can be minimized. We are encouraged by the experience we have gained over the past several years in performing these complex laparoscopic cases. The development of some of the newer surgical technologies has shortened operative times. Several recent studies have demonstrated short-term advantages with the laparoscopic approach compared with standard open operations for mucosal ulcerative colitis. Thus, this new approach is a viable and valuable option that may be offered to some patients with mucosal ulcerative colitis. PMID- 14760465 TI - Laparoscopy for diverticulitis. AB - Although the literature on laparoscopic surgery for diverticulitis includes data on more than 1800 patients, the quality of the studies is insufficient to draw definitive evidence-based conclusions. Nonrandomized evidence suggests that laparoscopic resection for uncomplicated diverticulitis of the sigmoid may fare better than its conventional counterpart not only in short-term outcome (preservation of the abdominal wall, shorter disability), but also in the long term (decreased rates of late symptomatic small bowel obstruction). Five-year recurrence rates show that a laparoscopic or conventional access is unlikely to have an impact, provided that the oral bowel end is anastomosed to the proximal rectum rather than to the distal sigmoid. The superiority of laparoscopy should be proven by measuring health-related and patient-centered outcome rather than surrogate endpoints. Areas of concern include replacing a conventional resection with laparoscopic suture, drainage, and colostomy in patients with free perforation and peritonitis. The role of laparoscopic surgery should be limited to resection for uncomplicated diverticulitis of the sigmoid performed by adequately trained surgeons. Benefits can be expected with this procedure, provided that indications for surgery are not influenced by the mode of access and that postoperative complication rates remain within the range of that for traditional colorectal surgery. PMID- 14760466 TI - Laparoscopy for adhesions. AB - Intestinal and abdominal adhesions may be responsible for a variety of clinical conditions, including chronic recurrent small-bowel obstruction, acute small bowel obstruction, closed-loop bowel obstruction and, debatably, abdominal or pelvic pain. Experience in laparoscopic surgery has increased at a rapid pace, thus adhesions are no longer considered a contraindication to treatment of these conditions. In recent years, numerous publications have reported the feasibility, safety, and favorable outcome of laparoscopic intervention in various adhesion related conditions. As adhesions are the most common cause of recurrent or acute bowel obstruction, this review will focus on the laparoscopic management of these conditions and outline the technical considerations, indications, contraindications, and results. PMID- 14760467 TI - Laparoscopic stoma creation and closure. AB - Laparoscopic surgery has demonstrated advantages over conventional open procedures. Specifically, avoiding an abdominal incision and allowing the complete inspection of the abdominal cavity, as well as the ability to obtain a biopsy sample, are some of the reasons that made this method of stoma creation advantageous. The creation of stomas by laparoscopy is one of the simpler laparoscopic procedures and is associated with a shorter learning curve compared with other colorectal procedures. This section discusses the indications, methods, and complications associated with the laparoscopic creation of stomas. PMID- 14760468 TI - Management of rectal prolapse: the role of laparoscopic approaches. AB - Rectal prolapse is a lifestyle-altering disability which has been treated with over 100 surgical options. The specific goals of surgical management of full thickness rectal prolapse are to minimize the operative risk in this typically elderly population, eradicate the external prolapse of the rectum, improve continence, improve bowel function, and reduce the risk of recurrence. The theoretical advantages of a laparoscopic approach are to couple reductions in surgical morbidity and good post-operative outcome. Studies which compare the same laparoscopic and open surgical approach for rectal prolapse have demonstrated that laparoscopy confers benefits related to postoperative pain, length of hospital stay, and return of bowel function. Virtually every type of open transabdominal surgical approach to rectal prolapse has been laparoscopically accomplished. Current laparoscopic surgical techniques include suture rectopexy, stapled rectopexy, posterior mesh rectopexy with artificial material, and resection of the sigmoid colon with colorectal anastomosis, with or without rectopexy. The growing body of literature supports the concept that laparoscopic surgical techniques can safely provide the benefits of low recurrence rates and improved functional outcome for patients with full thickness rectal prolapse. PMID- 14760469 TI - Laparoscopy for benign disease: robotics. AB - Currently available robotic surgical systems appear to be particularly suited for use in benign diseases of the gastrointestinal system. Minimally invasive operations for foregut conditions, such as gastroesophageal reflux disease and achalasia, require excellent visibility and precise tissue dissection. Benign lower gastrointestinal diseases, including inflammatory bowel disease and diverticulitis, also can be approached using robotic assistance. Disadvantages include expense and the loss of tactile feedback. Early clinical results are promising. PMID- 14760470 TI - Telementoring. AB - Telementoring began in the 1950s and is an advanced application of telemedicine that involves the removed guidance of a procedure where the student has no or limited experience. In the past 10 to 15 years, telemedicine has been revisited as a result of the healthcare delivery crisis, budgetary concerns, and the impact of managed care. In recent years, telementoring has had a number of successes which have led to further recent telementoring investigations and developments. Telementoring programs were established because it was impractical for specialized minimally invasive surgeons to proctor fellow surgeons during the adoption phase of new techniques. This catalyzed the establishment of formal telementoring procedural guidelines and networks. Efforts have been made in the remote direction of laparoscopic spermatic vein ligations, renal biopsy, nephrectomy, varicocelectomy, fetoscopy, and ophthalmology. Pilot studies in 2000 have statistically validated that telementoring can be as effective as on-site mentoring. In order to successfully conduct telementoring missions, however, it is important to follow a precise algorithm. If a standardized protocol is followed, it will ensure that telementoring is practiced safely and efficiently. PMID- 14760471 TI - Hand-assisted laparoscopic colectomy techniques. AB - Laparoscopic colon and rectal surgery has gained popularity over the last 10 years. The experience obtained from the more simple surgeries, such as cholecystectomy and appendectomy, and the development of better instruments have made colorectal standard laparoscopic surgery an easier procedure. However, the downside associated with this technique includes longer operative times, higher costs, loss of tactile sensation, and a two-dimensional view. In the last 5 years, a new type of instrument has appeared on the market: the hand-assist device. It gives back the tactile feeling and allows the surgeon to retrieve a three-dimensional evaluation of the abdomen. This instrument has changed the laparoscopic surgery field and permitted the expansion of laparoscopic colectomy to the most challenging and complex cases. Hand-assisted laparoscopic surgery seems to retain the same benefits as standard laparoscopic surgery and improve the operative time and the learning curve. This article reviews the benefits and the indications for the use of the hand-assist device, and the characteristics and types of hand-assist devices available and their instruction for use. It also focuses on the technical aspects of hand-assisted laparoscopic surgery. PMID- 14760472 TI - Catheter-directed thrombolysis for deep venous thrombosis. AB - Venous thromboembolism (VTE) represents a significant clinical problem, affecting patients of all age groups, nationalities, and socioeconomic strata. Despite its prevalence, the paradigms for care are largely centered around primary or secondary prophylaxis, with less emphasis on actual treatment of the thrombus. With the recent rapid development of advanced endovascular techniques, it is now feasible to dissolve many thrombi using catheter-directed thrombolysis (CDT), and favorable clinical experience has been reported in over 600 patients. If performed safely, the purported benefits of CDT for DVT include a decreased incidence of persistent phlebitic symptoms, improved quality of life and, possibly, a decreased incidence of recurrent thrombotic events. PMID- 14760473 TI - Vascular smooth muscle cell migration: current research and clinical implications. AB - Atherosclerosis and intimal hyperplasia are major causes of morbidity and mortality. These processes develop secondary to endothelial injury due to multiple stimuli, including smoking, diabetes mellitus, hypertension, and hyperlipidemia. Once this injury occurs, an essential element in the development of both these processes is vascular smooth muscle cell (VSMC) migration. Understanding the mechanisms involved in VSMC migration and ultimately the development of strategies by which this process can be inhibited, has been a major focus of research. The authors present a review of the extracellular proteins (growth factors, extracellular matrix components, and cell surface receptors) and intracellular signaling pathways involved in VSMC migration, as well as potential therapeutic approaches to inhibit this process. PMID- 14760474 TI - Associations between renovascular disease and prevalent cardiovascular disease in the elderly: a population-based study. AB - Atherosclerotic renovascular disease (RVD) is a suspected contributor to the morbidity and mortality of cardiovascular disease (CVD) through its potential effects on blood pressure and excretory renal function as well as through its associations with other forms of CVD. However, population-based data regarding the associations between the presence of RVD and prevalent CVD are lacking. The Cardiovascular Health Study (CHS) is a prospective, multicenter cohort study of CVD among elderly Americans. As part of an ancillary study, participants in the Forsyth County, North Carolina, cohort of the CHS were invited to undergo renal duplex sonography (RDS) to establish the presence or absence of RVD (defined as any focal peak systolic velocity >/= 1.8 m/second or the absence of a Doppler shifted signal from an imaged artery). Demographic, risk factor, and prevalent CVD data were obtained from the CHS coordinating center and matched with ancillary study participants. Eight hundred thirty-four CHS participants (including 525 women [63%], 309 men [37%], 194 African-Americans [23%], and 635 Caucasians [76%]) with a mean age of 77.2 +/-4.9 years underwent RDS examination. RVD was present in 57 participants (6.8%). Overall, clinical and/or subclinical manifestations of CVD were present in 603 participants (72.3%) at the time of RDS. Participants with RVD demonstrated a significantly greater prevalence of angina (p = 0.002), previous myocardial infarction (p < 0.001), >/= 25% diameter reducing internal carotid artery stenosis (p = 0.010), increased carotid intimal medial thickness (p = 0.003), and major electrocardiographic abnormalities (p = 0.013). Following adjustment for demographics and cardiovascular risk factors, the presence of RVD demonstrated a significant and independent association with prevalent coronary artery disease but not with prevalent cerebrovascular or lower extremity vascular disease. These results suggest important population-based associations between RVD and both clinical and subclinical manifestations of CVD, especially coronary artery disease. PMID- 14760475 TI - Hypothermia is an independent predictor of mortality in ruptured abdominal aortic aneurysms. AB - Hypothermia is known to significantly increase mortality in trauma patients, but the effect of hypothermia on outcomes in ruptured abdominal aortic aneurysms (RAAA) has not been evaluated. The authors reviewed their experience from 1990 to 1999 in 100 consecutive patients who presented with RAAA and survived at least to the operating room for surgical treatment. There were 70 men and 30 women, with a mean overall age of 74 +/-8 years. Overall mortality was 47%. Univariate ANOVA (analysis of variants) showed significant correlation with mortality for decreased intraoperative temperature, decreased intraoperative systolic blood pressure, increased intraoperative base deficit, increased blood volume transfused, increased crystalloid volume (all p < 0.001); decreased preoperative hemoglobin (p = 0.015); and increased age (p = 0.026). Patient sex, initial preoperative temperature, preoperative systolic blood pressure, and operating room time were not correlated with mortality in the univariate analysis. Using these same clinical variables, multiple logistic regression analysis showed only 2 factors independently correlated with mortality: lowest intraoperative temperature (p = 0.006) and intraoperative base deficit (p = 0.009). The mean lowest temperature for survivors was 35 +/-1 degrees C and for nonsurvivors 33 +/ 2 degrees C (p < 0.001). When patients were grouped by lowest intraoperative temperature, those whose temperature was < 32 degrees C (n = 15) had a mortality rate of 91%, whereas patients with a temperature between 32 and 35 degrees C (n = 50) had a mortality rate of 60%. In the group that remained at or > 35 degrees C (n = 35) the mortality rate was only 9%. A nomogram of predicted mortality versus temperature was constructed from these data and showed that for temperatures of 36, 34, and 32 degrees C the predicted mortality was 15%, 49%, and 84%, respectively. The authors conclude that hypothermia is a strong independent contributor to mortality in patients with ruptured abdominal aortic aneurysms and that very aggressive measures to prevent hypothermia are warranted during the resuscitation and treatment of these patients. PMID- 14760476 TI - Survival of ruptured abdominal aortic aneurysms in the west of Ireland: do prognostic indicators of outcome exist? AB - Ruptured abdominal aortic aneurysm (RAAA) is a demanding vascular surgical problem and the cause of significant morbidity and mortality. The aim of this study was to identify prognostic factors that influence outcome. Over 6 years, 42 ruptured abdominal aortic aneurysms were operated on with a mean diameter of 7.2 cm. RAAA was defined as free intraperitoneal rupture. Data were collected retrospectively from hospital medical records. The male: female ratio was 8:1 and the mean age was 74 years (range 55-89). Fifteen were in hypovolemic shock and 27 patients were clinically stable. The perioperative mortality rate for the 15 shocked patients was 60% (9 patients) and the 1-year cumulative survival rate was 33%. The perioperative mortality rate for the 27 clinically stable patients was 40% (11 patients) and the 1-year cumulative survival rate was 56%. Survival curves were constructed for these groups to compare male versus female, age >/= 70 versus age < 70, shocked versus stable, and preoperative hemoglobin (Hb) 10. No patient with preoperative cardiac arrest survived more than 24 hours. With VassarStats, the confidence interval for age, gender, hemodynamic status, and preoperative Hb were calculated. The standard weighted mean analysis by ANOVA gave a p value of < 0.001. The overall 30-day mortality rate was 47% (20 of 42) and the 1-year mortality rate was 52% (22 of 42). Male patients over 70 years with RAAA in hypovolemic shock with low Hb have a higher 30-day mortality rate and few survive more than 1 year. The study suggests that each of these 4 parameters separately was not a strong prognostic indicator. Collectively, however, they strongly influence the prognosis of patients with RAAA. These findings strengthen the case for selective treatment for RAAA. PMID- 14760477 TI - A comparison of AneuRx aortic cuff and zenith distal flare exclusion of common iliac artery ectasia for endovascular aneurysm repair. AB - Stent-grafts are ideally terminated within the common iliac artery (CIA). However, CIA ectasia may require hypogastric artery occlusion, with stent-graft extension to the external iliac artery. Alternatively, the diameter of the distal stent-graft may be increased, or flared, to allow exclusion of the abdominal aortic aneurysm. This report details the authors' experience with this technique. Forty-one patients received bifurcated stent-grafts (BSG): 20 received an AneuRx device, and 21 received a Zenith device. CIA ectasia (diameter 15-25 mm) was treated with a distal flare of 2-4 mm greater than the CIA diameter. Patients were followed up with computed tomography scan at 1, 6, and 12 months. Statistical analysis was performed using ANOVA within groups and unpaired two tailed t test between groups. A p value of < 0.05 was considered significant. Eight of 20 patients (40%) (11 CIA) received an AneuRx device and 13/21 (62%) (17 CIA) received a Zenith device, with a distal flare. Values are (n) mean (mm) +/- SE. There were no deaths, endoleaks, migrations, or conversions to open repair. Follow-up mean was 24.7 and 20.6 (range 15-28) months for AneuRx and Zenith groups, respectively. In comparing initial and 12-month CIA diameters, AneuRx grafts 20 +/-0.8 vs 21.5 +/-1.0 were not significantly different, p = 0.2, nor was the same comparison for Zenith, 17 +/-0.5 vs 19.1 +/-0.4, significant, p = 0.57. At a mean follow-up of 12 months, distal flare of iliac limbs with either AneuRx or Zenith devices affords a seal for CIA ectasia and/or aneurysms complicating EVAR. PMID- 14760478 TI - Correlation between preoperative and postoperative duplex vein measurements of the greater saphenous vein used for infrainguinal arterial reconstruction. AB - Vein diameter measurements using B-mode Doppler ultrasound (US) are used to assess the greater saphenous vein (GSV) for bypass operations; a 2.5-3.0 mm diameter is suggested as a minimum. Preoperative measurements are made while the vein is in the low-pressure venous system. This may not reflect the distended diameter of a vein after placement in the arterial system. This study compares preoperative and postoperative GSV diameters to identify the degree of dilatation and the minimal size adequate for use in arterial bypass operations. The GSV of 11 patients undergoing an infrainguinal arterial bypass were assessed by utilizing Doppler US. Measurements were taken every 10 cm, for 70 cm, along the course of the GSV before and 4 weeks after operation. All segments showed a percent increase in diameter from the preoperative to postoperative time points; 10 cm, 38+/-; 20 cm, 31+/-; 30 cm, 16+/-; 40 cm, 26+/-; 50 cm, 23+/-; 60 cm, 28+/ ; and 70 cm, 22+/-. A Bonferroni post hoc analysis between the 2 time point means showed a significant increase in means for the 2 time points of 9.49 units (Bonf p value < 0.001). Preoperative vein segments were divided into 3 categories: 4.1 mm. All showed a significant increase over time. Preoperative diameter measurements of the vein may not reflect the final distended diameter after bypass. Preoperative vein diameters 5 days). The maximum tolerated dose (MTD) was R115777 100 mg twice daily and irinotecan 100 mg/m(2) weekly. Non-DLTs were primarily rash, fatigue, diarrhea, and neutropenia. R115777 demonstrated linear pharmacokinetics without interaction with irinotecan and achieved serum levels required for antitumor activity in vitro. CONCLUSIONS: Serum levels of R115777 exceeded those necessary for FTase inhibition in vitro without evidence of interaction with irinotecan. However, the MTD of R115777 in this study was lower than that obtained with an alternate schedule. Thus, further development of this schedule is not recommended. PMID- 14760510 TI - Vaccination of prostatectomized prostate cancer patients in biochemical relapse, with autologous dendritic cells pulsed with recombinant human PSA. AB - This study was conducted in prostate cancer patients in biochemical relapse after radical prostatectomy, to assess the feasibility, safety, and immunogenicity of therapeutic vaccination with autologous dendritic cells (DCs) pulsed with human recombinant prostate-specific antigen (PSA) (Dendritophage-rPSA). Twenty-four patients with histologically proven prostate carcinoma and an isolated postoperative rise of serum PSA (>1 ng/ml to 10 ng/ml) after radical prostatectomy were included. The patients received nine administrations of PSA loaded DCs by combined intravenous, subcutaneous, and intradermal routes over 21 weeks. Postbaseline blood tests were performed at months 1, 3, 6, 9, and 12 (PSA levels), at months 6 and 12 (circulating prostate cancer cells), at month 6 (anti PSA IgG and IgM antibodies), and at up to eight time points before, during, and after immunization (PSA-specific T cells). Circulating prostate cancer cells detected in six patients at baseline were undetectable at 6 months and remained undetectable at 12 months. Eleven patients had a postbaseline transient PSA decrease on one to three occasions, predominantly occurring at month 1 (7 patients) or month 3 (2 patients). Maximum PSA decrease ranged from 6% to 39%. PSA decrease on at least one occasion was more frequent in patients with low Gleason score ( p=0.016) at prostatectomy and with positive skin tests at study baseline ( p=0.04). PSA-specific T cells were detected ex vivo by ELISpot for IFN gamma in 7 patients before vaccination and in 11 patients after vaccination. Of the latter 11 patients, 5 had detectable T cells both before and during the vaccination period, 4 only during the vaccination period, while 2 patients could for technical reasons not be assessed prevaccination. No induction of anti-PSA IgG or IgM antibodies was detected. There were no serious adverse events or otherwise severe toxicities observed during the trial. Immunization with Dendritophage-rPSA was feasible and safe in this cohort of patients. An immune response specific for PSA could be detected in some patients. A notable effect was the disappearance of circulating prostate cells in all patients who were RT PCR positive before vaccination. PMID- 14760512 TI - Suppression of GABA(B) receptor function in vivo by disulfide reducing agent, DL dithiothreitol (DTT). AB - RATIONALE: A recent in-vitro study demonstrated that the potent disulfide reducing agent, DL-dithiothreitol (DTT), may alter the structural stability of the GABA(B) receptor, probably inactivating the disulfide bonds between four cysteine residues located in the GABA(B1(a)) receptor structure. OBJECTIVES: The present study was designed to evaluate whether DTT treatment was capable of antagonizing some behavioral effects of pharmacological stimulation of the GABA(B) receptor. METHODS: Experiments on sedation/hypnosis induced by the GABA(B) receptor agonists baclofen, SKF 97541, CGP 44532 and gamma-hydroxybutyric acid (GHB) in DBA mice and selectively bred GHB-sensitive (GHB-S) rats, and a GHB drug discrimination study in Long Evans rats were conducted. Specificity of the DTT action on the GABA(B) receptor was investigated by assessing its effect on the sedative/hypnotic effect induced by diazepam, ketamine and ethanol. RESULTS: DTT prevented the sedative/hypnotic effect of all GABA(B) receptor agonists tested and also reversed baclofen-induced sedation/hypnosis. In contrast, DTT had no effect on, or even potentiated, sedation/hypnosis produced by diazepam, ketamine or ethanol. DTT completely blocked the discriminative stimulus effects of GHB. CONCLUSIONS: These results are discussed in terms of DTT altering the stability of the binding domain of the GABA(B) receptor, hindering the drug receptor interaction. PMID- 14760511 TI - Identification of a novel cytokine-like transcript differentially expressed in avian gammadelta T cells. AB - Chicken represents a species with a high frequency of gammadelta T cells in peripheral blood, suggesting an important function. To elucidate the genes specific for the avian gammadelta T cells, the suppression subtractive hybridization (SSH) between gammadelta and alphabeta T cells was used. The SSH library, which was successfully enriched for the TCR gamma and delta (both V and C region) sequences, provided gammadelta T-cell-specific genes, including, for example, the ribosomal proteins, signaling and structural molecules, and molecules related to transcription and translation. Among these genes, a clone named KK34 was shown to match the PFAM profile for IL-5 and to have 19.5% amino acid identity to the human interleukin 5 protein. Clone KK34 had lower sequence homology, from 5.4% to 15.6%, to other short-chain four-helix bundle superfamily members IL-3, IL-4, IL-13 and GM-CSF. The hydrophobic signal peptide sequence and the presence of cysteines needed for the interchain disulfide bonds were found to be conserved between clone KK34 and mammalian IL-5 proteins. Clone KK34 transcript expression was studied by RT-PCR, Northern blotting and in situ hybridization and it was confirmed to be expressed in avian gammadelta T cells. We propose that this clone, KK34, may represent the first nonmammalian IL-5, supporting the findings that gammadelta T cells are important in the development of allergy. PMID- 14760513 TI - Social instability in female rats: effects on anxiety and buspirone efficacy. AB - RATIONALE: Buspirone produces inconsistent effects in laboratory rodents. Individual housing increases the efficacy of buspirone in male rats, suggesting that the effects of this (and other) compounds become conspicuous in animals showing anxiety-like states. The effect of individual housing was, however, weak, and evident only when the locomotor suppressive effects of buspirone dissipated (i.e. 4 h after treatment). OBJECTIVES: The effects of social instability, a recently developed model of social stress in female rats, was investigated on both anxiety and the anxiolytic efficacy of buspirone. METHODS: Female rats were exposed to alternate days of isolation and moderate crowding for 2 weeks. Group composition was changed for each crowding phase. Basal anxiety and the anxiolytic efficacy of buspirone were assessed by the social interaction test of anxiety 24 h after the last crowding phase. RESULTS: Crowding appeared stressful, as it increased plasma glucocorticoid levels in less than 1 h. Anxiety-like behaviours were increased by social instability compared with stable group housing. In group housed controls, buspirone markedly suppressed locomotion, without clear effects on anxiety-related behaviours. Social instability attenuated the locomotor suppressive effects of buspirone, but made the anxiolytic effects of the compound more conspicuous. The effects of individual housing (assessed earlier) and social instability (assessed here) on buspirone efficacy appear qualitatively different. CONCLUSIONS: Buspirone abolishes stress-induced anxiety, but has no anxiolytic effects in controls. This is consistent with clinical findings, as the drug decreases anxiety in anxious patients but not in healthy humans. Laboratory models involving stress-induced anxiety-like states can improve our understanding of drug effects and efficacy. PMID- 14760514 TI - The influence on cognition of the interactions between lecithin, carnitine and carbohydrate. AB - It is accepted that acetylcholine-mediated neurones modulate memory. As lecithin, carnitine and glucose all influence acetylcholine metabolism, the possibility of synergistic interactions was considered. Four hundred young adult females randomly, and under a double-blind procedure, received capsules for 3 days that contained a placebo, lecithin (1.6 g/day), carnitine (500 mg/day) or carnitine plus lecithin. A battery of cognitive tests was administered prior to taking the capsules, after 3 days of taking the supplements, and for a third time after consuming either a glucose drink or a placebo. Reaction times were more rapid when carnitine and a glucose drink were taken together. Memory was enhanced in those taking a glucose rather than placebo drink. Neither mood nor the ability to sustain attention were influenced by these procedures. The hypothesis that memory would be facilitated by offering supplements of lecithin, carnitine and glucose was not supported. PMID- 14760515 TI - Suicidality and second generation antipsychotics in schizophrenia patients: a case-controlled retrospective study during a 5-year period. AB - RATIONALE: Rates of attempted suicide for individuals with schizophrenia are approaching 30%. Attempted suicide is among the most potent predictors of subsequent suicide. Several studies suggest that suicide is more likely to occur in patients who are not being adequately treated or not being treated at all. An effort was made in the last decade to evaluate the antisuicide effects of pharmacological treatment in schizophrenia with emphasis on the role of the newer second-generation antipsychotics (SGA). OBJECTIVE: The aim of the present study was to assess in a large cohort of schizophrenia patients the effects of exposure to SGA on suicidality of patients suffering from schizophrenia or schizoaffective disorder. The study is a retrospective case-controlled evaluation over a 5-year period undertaken in a large university affiliated tertiary care psychiatric hospital. METHODS: Between January 1998 and December 2002, all records of admissions of schizophrenia or schizoaffective disorder patients (ICD-10) were assessed. Data as to age, gender, diagnosis, suicide attempt prior to admission, treatment with antipsychotic medication, dose and duration of treatment (mg daily, duration) with SGA was extracted from patients' files. All patients who had attempted suicide prior to admission were defined as the index group. The case-controlled group was comprised of the next admission of a patient suffering from schizophrenia (or schizoaffective disorder), matched for gender and age, who did not attempt suicide. RESULTS: Records of 756 patients (4486 admissions for said period) were analyzed (56.6% male, mean age 39.1+/-13.5 years). Amongst 378 patients who attempted suicide (index group), 16.1% were exposed to SGA while 37% were exposed in the control group (P=0.0001). The protective effect (odds ratio) of treatment by SGA was 3.54 (95%CI: 2.4-5.3). Risperidone was more frequently prescribed in the control group (54.3%) and had a larger effect-size than olanzapine (3.16 versus 1.76), although not statistically significant. Clozapine was prescribed only to a few patients. CONCLUSIONS: Schizophrenia patients exposed to both risperidone and olanzapine may gain protection from suicidality. The antisuicide effects seem to differ between SGAs. The long duration and large sample size support this finding, despite the retrospective nature of this study. PMID- 14760516 TI - Combination of bone mineral density and upper femur geometry improves the prediction of hip fracture. AB - Bone mineral density (BMD) measured by dual-energy X-ray absorptiometry (DXA) is the main determinant of the clinical evaluation of hip fracture risk. However, it has been shown that BMD is not the only predictive factor for hip fracture, but that bone geometry is also important. We studied whether the combination of bone geometry and BMD could further improve the determination of hip fracture risk and fracture type. Seventy-four postmenopausal females (mean age 74 years) with a non pathologic cervical or trochanteric hip fracture without previous hip fracture or hip surgery constituted the study group. Forty-nine had a cervical fracture (mean age 73 years) and 25 had a trochanteric fracture (mean age 76 years). The control group consisted of 40 age-matched females (mean age 74 years). The geometrical parameters were defined from plain anteroposterior radiographs, and the potential sources of inaccuracy were eliminated as far as possible by using a standardized patient position and calibrated dimension measurements with digital image analysis. BMD was measured at the femoral neck (FEBMD), Ward's triangle (WABMD), and the trochanter (TRBMD). Stepwise linear regression analysis showed that the best predictor of hip fracture was the combination of medial calcar femoral cortex width (CFC), TRBMD, neck/shaft angle (NSA), and WABMD ( r=0.72, r(2)=0.52, P<0.001). The area under the receiver operating characteristic curve (ROC) for this model was 0.93, while the area under ROC for TRBMD alone was 0.81. At a specificity of 80%, sensitivity improved from 52.5% to 92.5% with this combination compared with TRBMD alone. The combined predictors of cervical and trochanteric fracture differed, being NSA, CFC, TRBMD, and WABMD for cervical and TRBMD and femoral shaft cortical thickness for trochanteric fracture. In addition, we found a statistically significant correlation between FEBMD and femoral shaft and femoral neck cortex width ( r=0.40, P<0.01 and r=0.30, P<0.01, respectively). The results confirm that the combination of BMD and radiological measures of upper femur geometry improve the assessment of the risk of hip fracture and fracture type compared to BMD alone, and that bone geometry plays an important role in the evaluation of bone strength. PMID- 14760517 TI - Radial bone density and breast cancer risk in white and African-American women. AB - A number of different models for assessing individual risk of breast cancer use known risk factors such as age, age at menarche, age at first live birth, previous breast biopsies, and family history. High bone mass in white women is also associated with an increased breast cancer risk; however, bone mass as a risk factor has not been studied in African-American women. We conducted a case control study to evaluate bone mineral density as a risk factor for breast cancer in white and African-American women. We recruited 221 women with newly diagnosed breast cancer from a comprehensive breast cancer center at a large university hospital, and 197 control women who were frequency matched for ethnicity and age. Odds ratios were based on proximal and distal radial bone density measured by peripheral bone densitometry (Norland pDEXA) and expressed as a standardized "Z score" (age and ethnicity specific). Logistic regression models were fitted controlling for body mass index, menopausal status, age, and HRT use (ever/never and duration). With proximal bone density Z-score included in the model as a continuous variable, a one-unit increase in radial shaft bone density increased the risk of breast cancer by 25% (p=0.02). When proximal bone density Z-score was analyzed as a dichotomous variable (< or = 0, > 0) the odds ratio was 1.98 (95% CI, 1.32 to 2.97); that is, having an above average proximal bone density (age specific) doubles the risk of breast cancer. There were no significant interactions with, and no appreciable confounding effects by, other covariates. An above-average radial shaft Z-score is a significant risk factor for breast cancer in both white and African-American women. The present study extends the association between bone mass and breast cancer risk to African-Americans, and suggests another potential application for bone density testing. PMID- 14760518 TI - Vitamin A intake and the risk of hip fracture in postmenopausal women: the Iowa Women's Health Study. AB - Excessive intake of vitamin A is postulated to have a detrimental effect on bone by inducing osteoporosis. This may lead to an increased risk of fracture, particularly in persons who are already at risk of osteoporosis. However, few studies have specifically examined the association of vitamin A intake through diet and supplement use, with fractures in a cohort of older, community-dwelling women. We prospectively followed a cohort of 34,703 postmenopausal women from the Iowa Women's Health Study to determine if high levels of vitamin A and retinol intake through food and supplement use were associated with an increased risk of hip or all fractures. A semiquantitative food frequency questionnaire was used to obtain the participants' baseline vitamin A and retinol intake. Participants were followed for a mean duration of 9.5 years for incident self-reported hip and nonhip fractures. After multivariate adjustment, it was revealed that users of supplements containing vitamin A had a 1.18-fold increased risk of incident hip fracture (n = 525) compared with nonusers (95% CI, 0.99 to 1.41), but there was no evidence of an increased risk of all fractures (n = 6,502) among supplement users. There was also no evidence of a dose-response relationship in hip fracture risk with increasing amounts of vitamin A or retinol from supplements. Furthermore, our results showed no association between vitamin A or retinol intake from food and supplements, or food only, and the risk of hip or all fractures. In conclusion, we found little evidence of an increased risk of hip or all fractures with higher intakes of vitamin A or retinol among a cohort of older, postmenopausal women. PMID- 14760519 TI - Bone mass and lifestyle related factors: a comparative study between Japanese and Inner Mongolian young premenopausal women. AB - The purpose of this study was to evaluate the ethnic difference in bone mass between Japanese and Inner Mongolian young premenopausal women and to assess the contribution of lifestyle related and anthropometric factors to bone mass. We studied 33 Japanese and 44 Inner Mongolian healthy young women, aged 20-34 years, in urban area. Speed of sound (SOS), broadband ultrasound attenuation (BUA) and stiffness index (SI) were measured at the calcaneus using quantitative ultrasound (QUS) analysis. Age at menarche, regularity of menstruation and lifestyle related factors were estimated by a self-reported questionnaire. There were no differences between the two groups in age, height, weight, BMI, regularity of menstruation, frequency of meat intake, frequency of yellow-green vegetable intake and exercise habit. Japanese women had significantly lower age at menarche and higher proportion of milk consumption habit at junior high school, senior school and present. Before adjustment, Japanese women had significantly higher SOS and SI than Inner Mongolian women. However, after adjustment for age at menarche and milk consumption habit at junior high school, both of which were significantly different between groups, no group-differences remained in either SOS or SI. These results suggest that the differences in age at menarche and milk consumption habit at junior high school, which relate to hormonal and nutritional status during puberty, may account for the differences in bone mass between Japanese and Inner Mongolian young women. PMID- 14760520 TI - Role of transposable elements in the propagation of minisatellites in the rice genome. AB - A survey of minisatellites (MSs) in 5.3 Mb of randomly selected rice DNA sequences from public databases was carried out to clarify the role of transposable elements (TEs) in the dispersal of MSs in the rice genome. The estimated frequency of MSs in this sample was one per 23.4 kb, and this frequency is approximately equivalent to that of Class I microsatellites in the rice genome. Of the MSs in the 5.3-Mb sequence sample, 82% were found to be present in multiple copies in the rice genome, and all of these were a part of TE sequences. In this study at least 61 TE groups were identified as MS carriers. It was also shown that the GC-rich MS pOs6.2H, which was previously reported to be one of the interspersed MSs in the rice genome, is a component of an En/Spm-like element. These results indicate that the majority of MSs in the rice genome are maintained in TEs, and amplified and dispersed as components of the TEs. The G+C content of the multi-locus MS sequences reflected that of the TE sequences containing those MSs, but no obvious bias towards the high G+C content of DNA was observed. Single locus MSs also did not show any obvious bias towards the high G+C content of DNA in the rice genome. In this respect, the MSs in the rice genome are quite different from those in the human genome: in the latter, the majority of MSs show an obvious bias towards the high G+C content of DNA. PMID- 14760521 TI - The ethylene biosynthetic and perception machinery is differentially expressed during endosperm and embryo development in maize. AB - The maize endosperm undergoes programmed cell death late in its development so that, with the exception of the aleurone layer, the tissue is dead by the time the kernel matures. Although ethylene is known to regulate the onset of endosperm cell death, the temporal and spatial control of the ethylene biosynthetic and perception machinery during maize endosperm development has not been examined. In this study, we report the isolation of the maize gene families for ACC synthase, ACC oxidase, the ethylene receptor, and EIN2 and EIL, which act downstream of the receptor. We show that ACC oxidase is expressed primarily in the endosperm, and only at low levels in the developing embryo late in its development. ACC synthase is expressed throughout endosperm development but, in contrast to ACC oxidase, it is transiently expressed to a significantly higher level in the developing embryo at a time that corresponds with the onset of endosperm cell death. Only two ethylene receptor gene families were identified in maize, in contrast to the five types previously identified in Arabidopsis. Members of both ethylene receptor families were expressed to substantially higher levels in the developing embryo than in the endosperm, as were members of the EIN2 and EIL gene families. These results suggest that the endosperm and embryo both contribute to the synthesis of ethylene, and they provide a basis for understanding why the developing endosperm is especially sensitive to ethylene-induced cell death while the embryo is protected. PMID- 14760522 TI - Isolation and characterization of genomic and transcribed retrotransposon sequences from sorghum. AB - Reverse transcriptase sequences from both major classes of retrotransposons were amplified from sorghum genomic DNA, leaf mRNA and callus protoplast mRNA. Sequence analysis of clones derived from genomic DNA demonstrated the presence of a wide variety of copia-like and gypsy-like elements. Twenty-four families of copia-like elements were found, of which at least thirteen were expressed in callus protoplasts. Two families (containing forty-eight subfamilies) of gypsy like elements were discovered, both closely related to Huck of maize. At least twenty-seven of these subfamilies were expressed in callus protoplasts. Most of these elements were expressed at high levels in protoplasts derived from embryogenic callus, but expression of only a few was detected (at low levels) in leaves. Sequence divergence within individual families was quite high, and all relatedness profiles were consistent with vertical transmission of these elements. These data indicate that sorghum contains a large number and diversity of retrotransposons, and that some may be useful as transposon tagging systems in callus protoplasts. PMID- 14760523 TI - Cloning, expression and partial characterization of a gene encoding the S15a ribosomal protein of Taenia solium. AB - Ribosomes, ribosomal proteins (r-proteins), and messenger and transfer RNAs catalyze the synthesis of proteins in organisms. To understand and define the components involved in this event in Taenia solium, we isolated and characterized a T. solium cDNA encoding the basic ribosomal protein S15a (TsS15a). The TsS15a cDNA produces a protein with M(r) (relative molecular mass) 14,988, which contains 22.3% of basic amino acids. Analysis comparing TsS15a protein with other S15a r-proteins indicates that this protein is highly conserved. A recombinant TsS15a protein with similar M(r) was produced in bacteria. Antibodies against recombinant TsS15a react with a 15-kDa protein in extracts from all life stages of T. solium and from all helminths tested. Hybridization studies showed the presence of two genes encoding a mRNA of 0.5 kb. Moreover, the gene presents an intron of 30 bp. Our phylogenetic analysis using S15a r-proteins reproduced the topologies reported for 16/18S rRNA. PMID- 14760525 TI - The identification of free-living environmental isolates of amoebae from Bulgaria. AB - A survey was carried out in Bulgaria to determine the presence of free-living amoebae (FLA) from environmental sources. In 171 (61.1%) of 280 samples, isolates of Acanthamoeba with group II or III morphology, as well as Hartmannella spp. were recovered. Five isolates named "6" (artificial lake), Ep (lake), G2 (soil), R4* (river) and PK (spring water)--all exhibiting a highly efficient proliferation in axenic cultures--were subsequently cloned and subjected to molecular analyses for identification and genotyping In accordance with morphological findings, PCR-based analyses identified four isolates (6, Ep, G2, R4*) belonging to the genus Acanthamoeba. Confirmation of these findings was obtained by phylogenetic analysis using partial sequencing of the 18S rDNA (ASA.S1) Acanthamoeba-gene. Comparison of these sequences with corresponding regions from other Acanthamoeba strains available from GenBank sorted all four isolates into the sequence type group T4 that contains most of the pathogenic Acanthamoeba strains already identified. The fifth isolate (PK) exhibited morphological characteristics matching those of Hartmannella, and scored negative in the Naegleria fowleri and Acanthamoeba PCRs. PMID- 14760526 TI - Prevalence of parasite eggs (Strongyloides westeri, Parascaris equorum, and strongyles) and oocysts (Emeria leuckarti) in the feces of Thoroughbred foals on 14 farms in central Kentucky in 2003. AB - Prevalence of internal parasites was determined by fecal examination for eggs and oocysts in Thoroughbred foals in central Kentucky in 2003. Fecal samples were examined from 733 foals on 14 farms. This included 70 trips to the farms and a total of 2,346 fecal samplings. Monthly collection of fecal samples was begun for four farms in February, six in March, three in April, and one farm in May. Termination of the study for all farms was the end of July. A criterion was that the foals be at least 10 days old for initial samplings. If available, the same foals were sampled each time, in addition to foals born in the interim between farm visits. Ages of the foals for the complete study varied from 10 to 223 days. Prevalence (mean %) was determined for eggs of Strongyloides westeri (1.5%), Parascaris equorum (22.4%), and strongyles (27.6%) and for oocysts of Eimeria leuckarti (41.6%) in feces of foals. Foals had infections of S. westeri on six farms (42.9%), of P. equorum on 12 farms (86%), and of strongyles and E. leuckarti on all 14 farms (100%). PMID- 14760527 TI - Prophylaxis of port system-associated thromboses in advanced oncology patients using heparin flushing. AB - PURPOSE: Thromboses occur in connection with the use of venous port systems. Valid data on the instillation of heparin-based solutions in the lumen of the port system are lacking. METHODS: One hundred and seventy-three patients with malignancy from 19 centres who had participated in an observation study of subcutaneous thromboprophylaxis with dalteparin-Na (Fragmin P/-Forte) were analysed with a view to flushing the port systems and investigating any related influence on the occurrence of catheter-associated thromboses. RESULTS: All catheter-associated thromboses were seen in centres which used either no UFH, or UFH concentrations of up to 250 IU/ml (8/108; 7.4%). The rate of thrombosis rose to 10% (6/60) if no high-risk dose of dalteparin was applied subcutaneously. On the other hand, the rate of catheter-associated thromboses under the high-risk dose of dalteparin and/or a more highly concentrated instillation fluid, at 0.9% (1/113), was much lower. One haemorrhage from gastric ulcer occurred under the highest UFH concentration in the instillation fluid (2,500 IU UFH/ml). CONCLUSION: The results indicate that a concentration between 500 IU UFH/ml and 1,000 IU UFH/ml in the instillation solution, at the same time as high-risk prophylaxis with subcutaneous dalteparin for prevention of catheter-associated thromboses, is effective in patients with manifest tumour disease. The instillation of LMWH-based solutions at a concentration of approx 500 anti-Xa units/ml should be discussed as a pending issue. PMID- 14760528 TI - Excluded right pulmonary artery in a neonate. PMID- 14760529 TI - Sisters with alpha-mannosidosis and systemic lupus erythematosus. AB - Alpha-mannosidosis is an autosomal recessive disorder caused by deficiency of lysosomal alpha-mannosidase (LAMAN). Here, we report two sisters with alpha mannosidosis who developed systemic lupus erythematosus (SLE). The sisters were both homozygous for a one bp deletion within the LAMAN gene resulting in a truncated gene product. The coincidence of alpha-mannosidosis and SLE are discussed with regard to both clinical and molecular findings. CONCLUSION: alpha mannnosidosis may contribute to the onset of systemic lupus erythematosus in predisposed patients. PMID- 14760530 TI - The humoral immune response in viral heart disease: characterization and pathophysiological significance of antibodies. AB - Several lines of evidence suggest a viral infection as the initiating event for the development of myocarditis (MC). Especially enteroviruses like coxsackie B3 virus have been shown to induce MC in humans and strains of MC-prone mice after an infection. The further course of the disease is, however, determined not only by the viral infection but also by the host's immune system. Both the humoral and the cellular immune system can modify the extent of the damage caused by the disease. The humoral immune system mounts an anti-viral immune response immediately after the infection; however, during the course of the disease, autoantibodies against a variety of different autoantigens emerge. The epitopes recognized by the anti-viral antibodies and those of several autoantibodies have been identified using synthetic peptides. The human disease could be transferred into SCID mice using peripheral blood leukocytes of patients, suggesting a pathophysiological significance of the autoimmune reaction. However, the significance of the humoral immune responses needs to be tested in randomized, prospective studies using immunoadsorption of autoantibodies in patients with inflammatory cardiomyopathy. PMID- 14760531 TI - Cartilage canals in the chicken embryo: ultrastructure and function. AB - In this study the detailed morphology and the function of cartilage canals in the chicken femur are investigated. Several embryonic stages (e 13.5, 16, 19, and 20) are examined by means of light microscopy, electron microscopy (TEM), and immunohistochemistry (VEGF, type I and II collagen). Our results show that cartilage canals originate from the perichondrium and form a complex pattern. Two types of canals are distinguishable: shell canals and communicating canals. Shell canals are in the reserve zone and are arranged in successive layers. Communicating canals spring from the shell canals and pass down into the proliferative zone and into the hypertrophic zone. These canals are conical shaped and are orientated nearly in parallel to the long axis of the femur. Cartilage canals comprise venules, arterioles, capillaries (mature and immature), and undifferentiated mesenchymal cells. No canal wall in the sense of an epithelium is elaborated. VEGF is detected in both types of canals and macrophages are found at the end of the cartilage canals. We conclude that the growth factor stimulates angiogenesis and that the latter cells erode the matrix ahead of the canals and thus enable the advancement of the vessels. The results clearly show that the canal matrix differs from the remaining cartilage matrix. The canal matrix contains type I collagen, few type II collagen fibrils and proteoglycans are lacking. In contrast, in the cartilage matrix type II collagen and proteoglycans are abundant but no type I collagen is found. Communicating canals are surrounded by a distinct layer of type I collagen indicating that osteoid is formed around these canals. Hypertrophic chondrocytes label for type I collagen and it seemed possible that chondrocytes adjacent to the communicating canals differentiate into bone-forming cells. Our results provide evidence that cartilage canals are involved in nourishment of the cartilage as well as in the ossification process. PMID- 14760532 TI - A re-evaluation of the premaxillary bone in humans. AB - The discovery of the premaxillary bone (os incisivum, os intermaxillare or premaxilla) in humans has been attributed to Goethe, and it has also been named os Goethei. However, Broussonet (1779) and Vicq d'Azyr (1780) came to the same result with different methods. The first anatomists described this medial part of the upper jaw as a separate bone in the vertebrate skull, and, as we know, Coiter (1573) was the first to present an illustration of the sutura incisiva in the human. This fact, and furthermore its development from three parts:-(1) the alveolar part with the facial process, (2) the palatine process, and (3) the processus Stenonianus-can no longer be found in modern textbooks of developmental biology. At the end of the nineteenth and in the early twentieth century a vehement discussion focused on the number and position of its ossification centers and its sutures. Therefore, it is hard to believe that the elaborate work of the old embryologists is ignored and that the existence of a premaxillary bone in humans is even denied by many authors. Therefore this re-evaluation was done to demonstrate the early development of the premaxillary bone using the reconstructions of Felber (1919), Jarmer (1922) and data from our own observations on SEM micrographs and serial sections from 16 mm embryo to 68 mm fetus. Ossification of a separate premaxilla was first observed in a 16 mm embryo. We agree with Jarmer (1922), Peter (1924), and Shepherd and McCarthy (1955) that it develops from three anlagen, which are, however, not fully separated. The predominant sutura incisiva (rudimentarily seen on the facial side in a prematurely born child) and a shorter sutura intraincisiva argue in this sense. The later growth of this bone and its processes establish an important structure in the middle of the facial skull. Its architecture fits well with the functional test of others. We also focused on the relation of the developing premaxilla to the forming nasal septum moving from ventral to dorsal and the intercalation of the vomer. Thus the premaxilla acts as a stabilizing element within the facial skeleton comparable with the keystone of a Roman arch. Furthermore, the significance of the premaxillary anlage for the closure of the palatine was documented by a synopsis made from a stage 16, 10.2 mm GL embryo to a 49 mm GL fetus. Finally the growth of the premaxilla is closely related to the development of the human face. Abnormal growth may be correlated to characteristic malformations such as protrusion, closed bite and prognathism. Concerning the relation of the premaxillary bone to cleft lip and palate we agree with others that the position of the clefts is not always identical with the incisive suture. This is proved by the double anlagen of an upper-outer incisor in a 55 mm fetus and an adult. PMID- 14760533 TI - Fluoride-induced alterations of enamel structure: an experimental study in the miniature pig. AB - We studied the structural changes in the enamel of mandibular third molars of miniature pigs administered a daily oral dose of 2 mg NaF (approximately 0.9 mg of fluoride) per kg body weight (added to the feed) for 1 year. The treatment period covered most of the secretory stage and the entire post-secretory stage of amelogenesis of the M(3). The enamel of the molars from the fluoride-fed pigs appeared opaque and chalky, and the erupted portions were stained brown. The underlying histopathological change was a pronounced subsurface hypomineralization of the enamel beneath a thin surface rim of higher mineral content. This enamel hypomineralization was attributed to a fluoride-induced impairment of the process of enamel maturation. The most conspicuous finding in the fluorotic enamel was the presence of numerous pit-type hypoplastic defects, denoting a marked fluoride-induced disturbance also of the secretory stage of amelogenesis. Microradiography and scanning electron microscopy revealed an enhanced incremental pattern in the outer enamel of the fluorotic molars. Typically, the bottom of larger hypoplastic defects was underlain by a broad, grossly accentuated incremental line. Occurrence of larger hypoplasias was further associated with the presence of aprismatic enamel, the formation of which was attributed to a loss of the prism-forming (distal) portion of the Tomes' processes of secretory ameloblasts. The findings in the miniature pigs closely parallel earlier observations on fluorotic enamel of free-ranging deer and wild boar from fluoride-polluted areas. PMID- 14760534 TI - Correlation between patterns of DNA mismatch repair hmlh1 and hmsh2 protein expression and progression of dysplasia in intraductal papillary mucinous neoplasms of the pancreas. AB - Defective DNA mismatch repair results from genetic or epigenetic alterations that most frequently inactivate the genes hMLH1 and hMSH2. This is thought to promote tumourigenesis by accumulation of mutations in oncogenes and tumour suppressor genes. This pathway, first reported in colon cancer, has been recently demonstrated in a subgroup of sporadic pancreatic adenocarcinomas. Intraductal papillary-mucinous neoplasms of the pancreas are a special type of pancreatic tumours, characterised by a spectrum of morphological changes from mild to moderate and to non-invasive, and they may associate with adenocarcinoma. An immunohistochemical study of hmlh1 and hmsh2 protein expression was performed on 26 intraductal papillary-mucinous neoplasms. All tumours showed nuclear expression of hmlh1 and hmsh2 proteins. There were two distinctive patterns of protein expression on the basis of the location of cells expressing these markers: the "normal" pattern, observed mainly in adenoma and rarely in intraductal papillary-mucinous neoplasms with moderate dysplasia and the "dysplastic" pattern, frequently encountered in moderate dysplasia neoplasms, non invasive and invasive carcinomas. These findings suggest that defective DNA mismatch repair, due to inactivation of hMLH1 and hMSH2, does not play a significant role in the pathogenesis of intraductal papillary-mucinous neoplasms of the pancreas. Two patterns of protein expression were observed and were correlated with the progression of dysplasia in intraductal papillary mucinous neoplasms. PMID- 14760535 TI - The role of GRAS proteins in plant signal transduction and development. AB - GRAS proteins are a recently discovered family of plant-specific proteins named after GAI, RGA and SCR, the first three of its members isolated. Although the Arabidopsis genome encodes at least 33 GRAS protein family members only a few GRAS proteins have been characterized so far. However, it is becoming clear that GRAS proteins exert important roles in very diverse processes such as signal transduction, meristem maintenance and development. Here we present a survey of the different GRAS proteins and review the current knowledge of the function of individual members of this protein family. PMID- 14760537 TI - Creatinine normalization in biological monitoring revisited: the case of 1 hydroxypyrene. AB - OBJECTIVES: To compare the apparent urinary excretion rates of both creatinine and 1-hydroxypyrene (1-OHP) and to assess the value of creatinine normalization for both toxicokinetic analysis and the routine examination of workers. METHODS: All urine samples were collected from individuals who had been exposed to polycyclic aromatic hydrocarbons (PAHs), occupationally and non-occupationally, for at least 24 consecutive hours. Urinary creatinine and 1-OHP were determined. 1-OHP excretion rates were expressed either as a function of creatinine excretion rate or as unadjusted values. Theoretical relationships between creatinine normalized excretion of metabolites and body weight-adjusted inhaled dose were drawn for men with a constant body mass index. RESULTS: Creatinine excretion rate paralleled 1-OHP excretion rate. The plot of creatinine excretion rate-adjusted excretion rate of 1-OHP vs time led to smooth curves for determination of toxicokinetic parameters. Creatinine normalization was adequate, even for samples with a urinary creatinine concentration below 0.5 g/l or above 3 g/l. A theoretical analysis revealed that men weighing between 50 kg and 100 kg, exposed to a constant dose of a pollutant producing a urinary metabolite excreted by the same mechanism as creatinine, would exhibit a body weight-adjusted dose span of 2 with an accompanying creatinine-normalized metabolite excretion span of 2.23 fold. CONCLUSION: The kinetics of creatinine excretion parallels that of 1-OHP, and a creatinine excretion rate-normalized excretion rate of 1-OHP appears to allow for a better determination of the toxicokinetic parameters of 1-OHP urinary excretion. At least in the case of 1-OHP, creatinine normalization seems valid, even for very dilute or very concentrated urine samples. Finally, because creatinine normalization not only compensates for variable diuresis but also correlates better with the body weight-adjusted dose of the parent compound, it should be used in biological monitoring of exposure to (PAHs) pyrene and to other substances whose urinary biomarker excretion kinetics parallel that of creatinine. PMID- 14760536 TI - Current concept of pathophysiological understanding and natural course of ulcerative colitis. AB - INTRODUCTION: According to the current paradigm both ulcerative colitis (UC) and Crohn's disease (CD) result from a complex interplay of genetic susceptibility factors, environmental factors, alterations of the physiological intestinal flora and a defective regulation of the intestinal immune system. DISCUSSION: The objective of this review is to give an overview of these factors and mechanisms, including genetic, environmental and microbial factors, with special alterations of relevant cellular components of the intestinal immune system such as T cells, macrophages and epithelial cells will then be addressed. In addition, the most relevant animal model systems that have contributed to our current pathogenetic understanding will be introduced. Clinically, the natural course of UC with special reference to the risk of colorectal cancer will be addressed. CONCLUSION: The elucidation of pathomechanisms at the level of the intestinal immune system provides the potential for novel, effective treatment strategies. Best surgical management of patients with UC, however, still remains a challenge. PMID- 14760539 TI - Comparison of polymerase chain reaction and two urinary antigen detection kits for detecting Legionella in clinical samples. PMID- 14760540 TI - Gordonia polyisoprenivorans septicemia in a bone marrow transplant patient. PMID- 14760541 TI - Appearance of the root canal walls after preparation with NiTi rotary instruments: a comparative SEM investigation. AB - The aim of this study was to evaluate, in vitro, with scanning electron microscope (SEM), the appearance of root canal walls shaped by three different rotary NiTi techniques and one conventional manual technique in human extracted teeth. Four different instruments were used: K3, Hero 642, RaCe and K-file. Each sample was irrigated with 5 ml of 5% NaOCl and 5 ml of 3% H2O2 and EDTA, Rc-Prep (1 ml). Each sample was prepared for SEM observation and analyzed in the coronal, middle and apical third, comparing its aspect with a predefined scale of four different parameters: presence of smear layer, pulpal debris, inorganic debris and surface profile. The apical third showed significantly more pulpal and inorganic debris, smear layer and a high number of surface profile irregularities. No significant difference was found at the coronal, middle and apical thirds between manual and rotary techniques for inorganic debris, smear layer and surface profile. Much pulpal debris were found in the apical third for K3 and RaCe compared with Hero 642 and K-file. In conclusion, mechanical rotary techniques with NiTi instruments produced quite similar results compared with a conventional manual technique using K-files. The present study demonstrated that dentin and pulpal debris, the morphology of smear layer and surface profile were only partially influenced by the type of endodontic instruments. The apical third was the anatomical area with greater amount of debris and smear layer. PMID- 14760542 TI - Sleep management training for cancer patients with insomnia. AB - Insomnia is a common phenomenon in cancer patients; nevertheless, there are only a few intervention results published covering this topic. We examined the effects of a multi-modal psychological sleep management programme combining relaxation techniques, sleep hygiene, cognitive techniques and advice in stimulus control technique on various sleep and quality-of-life variables. We compared two intervention groups up to 6 months after treatment, one with progressive muscle relaxation (n=80), the other with autogenic training (n=71). A control group (n=78) received only a standard rehabilitation programme. It was a heterogeneous sample of adult patients (mean age 58 years) predominantly with breast, kidney or prostate cancer staying for 3 or 4 weeks in an oncological rehabilitation clinic. In comparison to the control group, the analysis of variance for repeated measures (R-MANOVA) showed significant improvements over time, indicating that intervention group participants benefited with moderate- or large-scale effects on sleep latency (p<0.001), sleep duration (p<0.001), sleep efficiency (p<0.001), sleep quality (p<0.001), sleep medication (p<0.05) and daytime dysfunction (p<0.05). In quality-of-life subscales, there was mainly improvement over time. This may indicate a benefit of the rehabilitation treatment in general. No evidence was found for any differences between the two intervention groups. The results suggest that the use of a multi-modal psychological sleep intervention could enhance various sleep parameters and well being of patients. The efficacy on quality of life is still under review. PMID- 14760543 TI - In memoriam: Morton Spinner, MD 1928-2003. PMID- 14760559 TI - Get involved and make a difference. PMID- 14760560 TI - Recognizing the dietetics profession. PMID- 14760561 TI - Networking: it's an art. PMID- 14760562 TI - Avoiding medical errors: JCAHO documentation requirements. PMID- 14760538 TI - EST and microarray analyses of pathogen-responsive genes in hot pepper (Capsicum annuum L.) non-host resistance against soybean pustule pathogen (Xanthomonas axonopodis pv. glycines). AB - Large-scale single-pass sequencing of cDNA libraries and microarray analysis have proven to be useful tools for discovering new genes and studying gene expression. As a first step in elucidating the defense mechanisms in hot pepper plants, a total of 8,525 expressed sequence tags (ESTs) were generated and analyzed in silico. The cDNA microarray analysis identified 613 hot pepper genes that were transcriptionally responsive to the non-host soybean pustule pathogen Xanthomonas axonopodis pv. glycines ( Xag). Several functional types of genes, including those involved in cell wall modification/biosynthesis, transport, signaling pathways and divergent defense reactions, were induced at the early stage of Xag infiltration. In contrast, genes encoding proteins that are involved in photosynthesis, carbohydrate metabolism and the synthesis of chloroplast biogenetic proteins were down-regulated at the late stage of Xag infiltration. These expression profiles share common features with the expression profiles elicited by other stresses, such as fungal challenge, wounding, cold, drought and high salinity. However, we also identified several novel transcription factors that may be specifically involved in the defense reaction of the hot pepper. We also found that the defense reaction of the hot pepper may involve the deactivation of gibberellin. Furthermore, many genes encoding proteins with unknown function were identified. Functional analysis of these genes may broaden our understanding of non-host resistance. This study is the first report of large scale sequencing and non-host defense transcriptome analysis of the hot pepper plant species. (The sequence data in this paper have been submitted to the dbEST and GenBank database under the codes 10227604-10236595 and BM059564-BM068555, respectively. Additional information is available at http://plant.pdrc.re.kr/ks200201/pepper.html). PMID- 14760563 TI - Insulin resistance syndrome: a potent culprit in cardiovascular disease. PMID- 14760564 TI - Readiness to implement Hazard Analysis and Critical Control Point (HACCP) systems in Iowa schools. AB - OBJECTIVE: To evaluate current food-handling practices, food safety prerequisite programs, and employee knowledge and food safety attitudes and provide baseline data for implementing Hazard Analysis and Critical Control Point (HACCP) systems in school foodservice. DESIGN: One member of the research team visited each school to observe food-handling practices and assess prerequisite programs using a structured observation form. A questionnaire was used to determine employees' attitudes, knowledge, and demographic information. SUBJECTS/SETTING: A convenience sample of 40 Iowa schools was recruited with input from the Iowa Department of Education. STATISTICAL ANALYSES: Descriptive statistics were used to summarize data. One-way analysis of variance was used to assess differences in attitudes and food safety knowledge among managers, cooks, and other foodservice employees. Multiple linear regression assessed the relationship between manager and school district demographics and the food safety practice score. RESULTS: Proper food-handling practices were not being followed in many schools and prerequisite food safety programs for HACCP were found to be inadequate for many school foodservice operations. School foodservice employees were found to have a significant amount of food safety knowledge (15.9+/-2.4 out of 20 possible points). School districts with managers (P=.019) and employees (P=.030) who had a food handler certificate were found to have higher food safety practice scores. APPLICATIONS/CONCLUSIONS: Emphasis on implementing prerequisite programs in preparation for HACCP is needed in school foodservice. Training programs, both basic food safety such as ServSafe and HACCP, will support improvement of food handling practices and implementation of prerequisite programs and HACCP. PMID- 14760565 TI - A camera's view of consumer food-handling behaviors. AB - OBJECTIVE: To compare consumer food-handling behaviors with the Fight BAC! consumer food-safety recommendations. DESIGN: Subjects were videotaped in their home while preparing a meal. Videotapes were coded according to Fight BAC! recommendations. A food-safety survey was administered and temperature data was collected. SUBJECTS/SETTING: A market research company randomly recruited subjects by telephone. Ninety-nine consumers participated (92 women, seven men). STATISTICAL ANALYSIS PERFORMED: Descriptive statistics were used. RESULTS: Overall, subjects did not follow the Fight BAC! recommendations for safe food handling. Handwashing was inadequate. The average hand wash length was significantly lower than the 20-second recommendation. Only one-third of subjects' hand wash attempts were with soap. Surface cleaning was inadequate with only one-third of surfaces thoroughly cleaned. Moreover, one-third of subjects did not attempt to clean surfaces during food preparation. Nearly all subjects cross-contaminated raw meat, poultry, seafood, eggs, and/or unwashed vegetables with ready-to-eat foods multiple times during food preparation. Unwashed hands were the most common cross-contamination agent. Many subjects undercooked the meat and poultry entrees. Very few subjects used a food thermometer. APPLICATIONS/CONCLUSIONS: Consumers make many food-handling errors during food preparation, increasing their risk of foodborne illness. Dietetics professionals need to familiarize themselves with the Fight BAC! consumer food-safety recommendations; understand where consumers are making food-handling errors; increase food safety awareness; and educate consumers, especially those in high risk populations, about safe food handling at home. PMID- 14760566 TI - A media literacy nutrition education curriculum for head start parents about the effects of television advertising on their children's food requests. AB - OBJECTIVE: To evaluate whether a media literacy nutrition education curriculum about the effects of television advertising on children's food choices influenced the behavior, attitudes, and knowledge of Head Start parents. SUBJECTS: Participants were a convenience sample of 35 parents from Head Start programs. DESIGN: This study used a pretest-posttest, comparison condition-intervention condition design. INTERVENTION: The 35 parents participated in both a four-week food safety curriculum (to serve as an educational placebo, comparison condition) that was followed immediately by a four-week media literacy nutrition education curriculum (intervention condition). MAIN OUTCOME MEASURES: Evaluation measures included parents' understanding of the persuasive techniques of commercials; ability to distinguish between truths and claims in advertising; and outcome expectations, values, self-efficacy, and behaviors in relation to talking about television advertisements with children while co-viewing or in response to purchase requests in the grocery store. STATISTICAL ANALYSES: Paired t tests, analysis of covariance, and chi(2) analyses were used. RESULTS: The media literacy nutrition education intervention curriculum had significant effects in terms of Head Start parents' understanding television advertising (P<.001), attitudes about television advertisements (P<.001), outcome expectations (P<.05), values (P<.01), self-efficacy (P<.001), and TV mediation behaviors (P<.001), and understanding of, and ability to read, food labels (P<.001). CONCLUSIONS/APPLICATIONS: Results suggest that a media literacy nutrition education curriculum can be easily conducted by dietitians. Dietitians can modify the curriculum to teach parents how to critically analyze many other forms of media (supermarket magazines, brochures, newspapers, Web sites) that sell nutrition misinformation to the public. PMID- 14760567 TI - Dietary intake in the lower Mississippi delta region: results from the Foods of our Delta Study. AB - OBJECTIVE: To collect and evaluate food intake data from a culturally diverse population and compare with national survey data. DESIGN: The Foods Of Our Delta Study was a baseline, cross-sectional survey that utilized random-digit dialing methodology to identify the sample. Food intake was obtained from a 24-hour dietary recall administered by computer-assisted telephone interview using the multiple-pass method. SUBJECTS/SETTING: One thousand seven hundred fifty-one adults and 485 children in the Lower Mississippi Delta (Delta) of Louisiana, Arkansas, and Mississippi. STATISTICAL ANALYSES PERFORMED: Comparisons of subsets within the Delta were made using weighted t tests. Comparisons of the Delta with the overall US population from the US Department of Agriculture Continuing Survey of Food Intakes by Individuals and with the Dietary Reference Intakes were made using independent sample z tests of weighted estimates. RESULTS: Energy intake did not differ between the Delta and the US populations. Intakes of protein were lower, fat higher, and certain micronutrients lower in Delta adults than in US adults. Delta adults had a 20% lower intake of fruits and vegetables than the US adults and generally poorer adherence to recommendations of the Food Guide Pyramid. African American Delta adults generally consumed less-optimal diets than white Delta adults. Delta children had diets similar to children of the Continuing Survey of Food Intakes by Individuals sample population, but lower intakes were noted for vitamins A, C, riboflavin, and B-6, and for calcium and iron. APPLICATIONS/CONCLUSIONS: Data such as these will help drive intervention development in this rural region and perhaps set the stage for research in similarly impoverished areas. PMID- 14760568 TI - Dietary patterns, smoking, and subclinical heart disease in women: opportunities for primary prevention from the Framingham Nutrition Studies. AB - OBJECTIVES: To investigate the relationship between a heart-healthy dietary pattern and subclinical heart disease in women, and to identify potential opportunities for primary prevention. DESIGN: Prospective analysis in which dietary patterns and cardiovascular disease (CVD) risk factors were assessed at baseline. Presence of subclinical heart disease was assessed using carotid atherosclerosis (stenosis >or=25%) measured by ultrasound at 12-year follow-up. SUBJECTS/SETTING: We studied 1,423 women in the population-based Framingham Offspring/Spouse (FOS) Study cohort, Framingham, Massachusetts. Subjects did not have CVD at baseline. STATISTICAL ANALYSES: CVD risk factor differences among the dietary clusters were evaluated using analysis of covariance and logistic regression. The relationship between heart-healthy and less heart-healthy dietary patterns and the presence of subclinical heart disease at follow-up was examined using odds ratios calculated from multivariate logistic regressions; stratification by smoking status (current, former, never) was also explored. RESULTS: Women who ate a heart-healthy diet had more favorable baseline CVD risk factor profiles. The age-adjusted odds of subclinical heart disease at follow-up was 40% lower for heart-healthy women (OR 0.60, P=.02). Multivariate adjustment for BMI, blood lipid levels, and blood pressure only slightly attenuated these odds. The odds remained reduced after adding pack-years of smoking to the multivariate model, but statistical significance was attenuated (OR 0.74, P=.20). In analyses stratified by smoking status, women who consumed a heart-healthy diet and who had never smoked had more than 80% less odds for subclinical heart disease compared with smokers whose diets were less heart-healthy (adjusted OR 0.17; P=.0001). CONCLUSIONS: Women who achieve a heart-healthy eating pattern, in combination with the avoidance of smoking, have a lower odds of subclinical heart disease. Among former smokers, the avoidance of smoking seemed to have somewhat more influence than diet on stenosis risk. A public health priority for women to promote the primary prevention of heart disease is the adoption of positive lifestyle behaviors, especially healthful eating (dietary patterns rich in fruits, vegetables, low-fat dairy foods, leaner protein sources, and lower in fats) and the avoidance of smoking. PMID- 14760569 TI - Attitudes, practices, and concerns about child feeding and child weight status among socioeconomically diverse white, Hispanic, and African-American mothers. AB - Parents play an important role in the development of their children's eating behaviors. We conducted 12 focus groups (three white, three African-American, and three Hispanic-American low-income groups; three white middle-income groups) of mothers (N=101) of 2- to less than 5-year-old children to explore maternal attitudes, concerns, and practices related to child feeding and perceptions about child weight. We identified the following major themes from responses to our standardized focus group guide: 12 groups wanted to provide good nutrition, and most wanted children to avoid eating too many sweets and processed foods; 12 groups prepared foods their children liked, accommodated specific requests, and used bribes and rewards to accomplish their feeding goals (sweets were commonly used as bribes, rewards, or pacifiers); and 11 of 12 groups believed their children were prevaricating when they said they were full and mothers encouraged them to eat more. The common use of strategies that may not promote healthful weight suggests work is needed to develop culturally and socioeconomically effective overweight prevention programs. Further study is needed to verify racial/ethnic or income differences in attitudes, practices, and concerns about child feeding and perceptions of child weight. PMID- 14760570 TI - Dietary intake of children at high risk for cardiovascular disease. AB - Cardiovascular disease and obesity begin in childhood, and dietary interventions to prevent them should be initiated then. We hypothesized that children who were at high risk for cardiovascular disease based on family history would have diets that were different than those of children from low-risk families. Two hundred ninety-seven children were screened for family history of early cardiovascular disease; had height, weight, and finger-stick total cholesterol measured; and filled out food frequency questionnaires. Sixty-eight (23%) children were at risk for cardiovascular disease. Cholesterol was significantly higher compared with those not at risk (4.71+/-0.93 mmol/L vs 4.35+/-0.92 mmol/L, P=.005). Intakes of energy, fat, fiber, and cholesterol were similar between groups. Children at high risk for cardiovascular disease were no more likely to meet guidelines for heart healthy diets than were children at low risk. Families need guidance to change dietary patterns to prevent future disease. PMID- 14760571 TI - Systematic review of literature on the cost-effectiveness of nutrition services. AB - Employers and health plan directors would like to know whether it is cost effective to include outpatient nutrition services as a covered benefit. The purpose of this systematic review was to examine the strength of evidence on the cost-effectiveness of outpatient nutrition services from an economic perspective. All randomized controlled trials published between January 1966 and September 2001 that reported on costs and effectiveness of outpatient nutrition services for any indicated condition were identified and reviewed. Paired reviewers abstracted data from and assessed the quality of each eligible randomized controlled trial; 13 studies met the eligibility criteria. Relatively consistent evidence exists to support the cost-effectiveness of nutrition services in the reduction of serum cholesterol levels (eg, 20 dollars to 1,268 dollars per mmol/L decrease in serum low-density lipoprotein level), weight loss (2.40 dollars to 10 dollars per pound lost), and blood glucose (5 dollars per mmol/L decrease), and for target populations with diabetes mellitus and hypercholesterolemia. However, the randomized controlled trials had important limitations and used different cost perspectives. Limited evidence of economic benefit exists to support coverage of outpatient nutrition services for selected indications. More randomized controlled trials of nutrition services should be conducted, taking into consideration all potential candidates for nutrition therapy and all potential costs to patients, providers, and payers. PMID- 14760572 TI - Adolescents' attitudes about and consumption of low-fat foods: associations with sex and weight-control behaviors. AB - The objective of the study was to examine adolescents' attitudes about and consumption of low-fat foods by sex and weight-control behaviors. Ninth- to 12th grade students (n=1,083) in 20 Minnesota high schools completed mail surveys (response rate=74%) as part of baseline measurements (Fall 2001) in a randomized controlled trial (TACOS: Trying Alternative Cafeteria Options in Schools). Linear models examined attitudes and consumption of low-fat foods by sex and weight control behaviors. Girls were more likely than boys to report positive attitudes and consumption of low-fat foods (all P<.01). Weight-control practices were associated with more positive attitudes about and higher consumption of low-fat foods. Promoting low-fat snacks to adolescents who are interested in weight control may be an effective component of nutrition intervention programs because these students have more positive attitudes about low-fat foods. Dietitians' efforts to educate adolescents about the benefits and healthfulness of low-fat foods would aid intervention programs. PMID- 14760573 TI - Food security status of households in Appalachian Ohio with children in Head Start. AB - This study measured food security and hunger of households involved in Head Start in a rural Appalachian county and assessed factors that could affect food security and hunger. A convenience sample of households with children enrolled in the Head Start program in Athens County, Ohio, were sampled (n=710), with adults from 297 (42%) households responding. The survey instrument included the 18 question US Household Food Security Survey Module for measuring hunger and food insecurity. Of those responding, 152 households (51.2%) were food secure and 145 (48.8%) were food insecure. Ninety (30.3%) had experienced hunger in the previous 12 months, and 41 (13.8%) households were classified as food insecure with childhood hunger. Hunger was related to a variety of household characteristics and associated with several factors, including participation in food banks, dependence on family members and friends outside of the household for food, lacking reliable transportation, and not having a garden. PMID- 14760574 TI - Nutrient intake of Korean-American, Korean, and American adolescents. AB - This study compared dietary intakes of Korean, Korean-American, and American adolescents. Data were collected by 24-hour recall method from 471 Korean and 134 Korean-American adolescents. Data for American adolescents were obtained from NHANES III. Korean Americans had the lowest energy and cholesterol intakes but a higher percentage of energy from fat compared with Koreans. Korean and Korean Americans had lower intakes of calcium, iron, and zinc but higher sodium intake than Americans. Intakes of energy, folate, calcium, and iron in all three groups were below the recommended intakes for American adolescents. Korean Americans consumed cooked rice and Kimchi less often, but cookies, sweets, and soda were consumed more frequently than Koreans. These results indicate that Korean American adolescents' dietary profiles are midway between those typical in Korea vs the United States. Nutrition education must focus on the benefits and weaknesses of Korean and western diets and how to blend the two cultures for optimal nutrition. PMID- 14760575 TI - Intercollegiate student athlete use of nutritional supplements and the role of athletic trainers and dietitians in nutrition counseling. AB - A survey of university student athletes was conducted to determine supplement use, perceived efficacy of supplements, availability and use of nutrition services, and perceived nutrition knowledge of athletic trainers. Results from 236 athletes showed that 88% used one or more nutritional supplements, yet perceived efficacy was moderate (2.9 or less; 5-point scale). Classes (69.4%), brochures (75%), and individual counseling (47%) were available and were used by 29.9%, 33.2%, and 17.9% of athletes, respectively. Primary sources of nutrition information were athletic trainers (39.8%), strength and conditioning coaches (23.7%), and dietitians (14.4%). Athletes perceived athletic trainers to have strong nutrition knowledge (mean=3.8+/-0.9; 5-point scale). Many (23.5%) did not know whether a dietitian was available. Dietitians must accelerate their marketing efforts to student athletes, work closely with athletic trainers to provide sound nutrition information, and provide services that meet the needs of a diverse population of student athletes. PMID- 14760576 TI - Children who avoid drinking cow's milk are at increased risk for prepubertal bone fractures. AB - The full fracture histories of 50 children (30 girls and 20 boys, age range 3 to 13 years) who had avoided drinking cow's milk for prolonged periods were compared with those in a birth cohort of more than 1,000 children from the same city. Children who avoided milk did not use calcium-rich food substitutes appropriately and had low dietary calcium intakes and low bone mineral density values. Many were overweight (22 of 50). Significantly more of the children who avoided milk reported fractures (16 observed vs 6 expected, chi(2)=31.0, P<.001, df=5). They also experienced more total fractures than the birth cohort population (22 observed vs 8 expected, chi(2)=33.6, P<.001, df=5). All of the fractures occurred before puberty, the majority (18 of 22) being associated with only slight trauma. Forearm fractures were especially common (12 fractures). We conclude that young children avoiding milk are prone to fracture. PMID- 14760578 TI - Position of the American Dietetic Association: use of nutritive and nonnutritive sweeteners. AB - Sweeteners elicit pleasurable sensations with (nutritive) or without (nonnutritive) energy. Nutritive sweeteners (eg, sucrose, fructose) are generally recognized as safe (GRAS) by the Food and Drug Administration (FDA), yet concern exists about increasing sweetener intakes relative to optimal nutrition and health. Dietary quality suffers at intakes above 25% of total energy (the Institutes of Medicine's suggested maximal intake level). In the United States, estimated intakes of nutritive sweeteners fall below this, although one in four children (ages 9 to 18 years) can surpass this level. Polyols (sugar alcohols), GRAS-affirmed or petitions filed for GRAS, add sweetness with reduced energy and functional properties to foods/beverages and promote dental health. Five nonnutritive sweeteners with intense sweetening power have FDA approval (acesulfame-K, aspartame, neotame, saccharin, sucralose) and estimated intakes below the Acceptable Daily Intake (level that a person can safely consume everyday over a lifetime without risk). By increasing palatability of nutrient dense foods/beverages, sweeteners can promote diet healthfulness. Scientific evidence supports neither that intakes of nutritive sweeteners by themselves increase the risk of obesity nor that nutritive or nonnutritive sweeteners cause behavioral disorders. However, nutritive sweeteners increase risk of dental caries. High fructose intakes may cause hypertriglyceridemia and gastrointestinal symptoms in susceptible individuals. Thus, it is the position of The American Dietetic Association that consumers can safely enjoy a range of nutritive and nonnutritive sweeteners when consumed in a diet that is guided by current federal nutrition recommendations, such as the Dietary Guidelines for Americans and the Dietary References Intakes, as well as individual health goals. Dietetics professionals should provide consumers with science-based information about sweeteners and support research on the use of sweeteners to promote eating enjoyment, optimal nutrition, and health. PMID- 14760579 TI - Position paper update for 2004. PMID- 14760580 TI - Mainstreaming international outcomes research in dietetics. PMID- 14760581 TI - Meeting the challenges of dietetics practice with evidence-based decisions. PMID- 14760589 TI - Routine measurement of radioisotope left ventricular ejection fraction prior to vascular surgery: is it worthwhile? AB - OBJECTIVE: To determine whether estimation of left ventricular (LV) ejection fraction (EF) by means of multiple gated acquisition (MUGA) scanning could reliably stratify cardiac risk prior to elective major vascular surgery. METHODS: A review of the English-language literature. RESULTS AND CONCLUSIONS: Twenty-two studies enrolling a total of 3096 patients were identified from 1984 to date. Selection bias, blinding of the results, different cut-off limits, and several retrospective studies were some of the problems preventing a comprehensive analysis. The resting LVEF was not found to be a consistent predictor of perioperative ischaemic cardiac events. In the perioperative phase, poor LV function was, mainly, predictive of congestive heart failure, and, in the long term, of cardiac outcome. The presence of myocardial wall motion abnormalities was also associated with both a higher chance of postoperative cardiac complications and a worse long-term cardiac outcome. Although measurements of LV function seem to play a key role in defining a patient's long-term prognosis, the value of routinely measuring LVEF preoperatively is limited and, therefore, MUGA scanning cannot be recommended as a general screening test. Despite this, it has been widely used for cardiac risk assessment in vascular surgery, and only recently its popularity has started declining. Other tests, such as stress echocardiography and myocardial perfusion imaging, used selectively in moderate risk patients can refine prediction of cardiac risk. In the future, gated stress myocardial perfusion scintigraphy, perhaps combined with ANP/BNP plasma level determination, may become a first choice test in preoperative cardiac risk assessment. PMID- 14760590 TI - Economy class stroke syndrome: case report and review of the literature. AB - OBJECTIVES: Venous thromboembolism associated with travelling, or economy class syndrome, is increasingly recognised as a sequence of long haul flights and so paradoxical cerebral embolism through a patent foramen ovale. Materials and methods. We present a new case of economy class stroke syndrome and review of the literature using MEDLINE search. RESULTS: Literature review identified 12 additional cases. In most of them, stroke occurred in close approximation with landing of the aircraft following a long-haul flight. Venous thromboembolism was present in 58%, while a patent foramen ovale was diagnosed with contrast echocardiography in all but one case. Our case presented with severe left hemispheric stroke, and significant delay, two days after a long-haul flight. CONCLUSIONS: The small number of reported cases indicates either the rarity of this entity or unawareness of its existence. The true incidence of this condition remains unknown. However, because of treatment implications such as the need to treat venous thromboembolism or close the patent foramen ovale, clinicians should be aware of this entity. PMID- 14760591 TI - Immunocytochemical study on endothelial integrity of saphenous vein grafts harvested by minimally invasive surgery with the use of vascular mayo stripers. A randomized controlled trial. AB - OBJECTIVES: The aim of this study is to compare the endothelial integrity of saphenous vein grafts harvested by minimally invasive surgery and veins harvested conventionally for coronary artery bypass surgery in 200 participants who were assigned to interventions by using random allocation. DESIGN: Randomized controlled trial. Methods. Immunocytochemistry with anti-CD 31 antibodies and anti-nitric oxide synthase (NOS) antibodies were employed to identify the endothelial integrity. RESULTS: The CD 31 immunostaining showed that the endothelial cell integrity of the minimally invasive harvested veins was preserved in 82+/-13% of the circumference of luminal endothelium, while in conventionally harvested grafts it was reduced to 64+/-15% (p=0.05).> This was associated with the lack of CD 31 expression in vasa vasorum (10 and 18%) in both groups, respectively, (p=0.02). The NOS immunostaining revealed that the endothelial integrity of the minimally invasive harvested grafts was preserved in 96+/-4% of the luminal endothelium circumference as compared to 74+/-10% in conventionally harvested grafts (p=0.05). The percentage of cases with the lack of NOS expression in all vasa vasorum was 12 and 21%, in G1 and G2, respectively, (p=0.02). CONCLUSION: The endothelial integrity of saphenous vein grafts harvested by minimally invasive surgery is better preserved than with the grafts obtained by the conventional manner. This could play an important role in improving vein graft patency rates. PMID- 14760592 TI - Bovine mesenteric vein graft (ProCol) in critical limb ischaemia with tissue loss and infection. AB - OBJECTIVES: Poor results have been reported following infrainguinal reconstructions using heterogenous grafts. The objective of this study was to assess the use of bovine mesenteric vein (ProCol) graft in patients with critical limb ischaemia (CLI), tissue loss/infection and no autologous vein available for reconstruction. METHODS: Prospective analysis of 32 patients with CLI and tissue loss/infection, in whom reconstruction with ProCol was undertaken between October 1999 and May 2002. RESULTS: The primary patency rate was 16% at 1 month. After thrombectomy, the secondary patency rate was 50% at 1 month and 26% at 14 months. No graft infections were seen. Aneurysmal dilatation of the graft occurred in 2 (6%). Limb salvage at 14 months was 47%. CONCLUSION: In patients with critical limb ischaemia, tissue loss/infection and no available vein, the ProCol graft may be an alternative. However, primary patency is a problem. In situations without tissue loss/infection, where the risk of graft infection is less, prosthetic material may be a better alternative. PMID- 14760593 TI - Outcome of conservative therapy of patients with severe intermittent claudication. AB - BACKGROUND: Intermittent claudication due to peripheral artery disease (PAD) can be treated conservatively, or by revascularization. OBJECTIVES: To assess the short-term outcome of conservatively-treated claudicants, and determine predictors for clinical improvement. Design. A retrospective cohort study. METHODS: We evaluated Fontaine stage, walking distance and ankle brachial index (ABI) at baseline and after median 9 months (interquartile range (IQR) 6-24) in 181 patients with severe claudication. RESULTS: We found clinical improvement by at least one Fontaine stage in 38 patients (21%) with an increased walking distance from baseline median 100 m (IQR 50-150) to follow-up median 650 m (IQR 300 to unlimited; p<0.001), but without changes in ABI (median 0.57, IQR 0.48 0.73 vs. median 0.54, IQR 0.45-0.81; p=0.10). One hundred and thirty-eight patients (76%) remained clinically and hemodynamically stable. A worsening of the clinical stage but without amputation was recorded in five patients (3%). Female gender (hazard ratio (HR) 0.51, p=0.052), diabetes (HR 0.35, p=0.020), and baseline ABI below 0.44 (HR 0.31, p=0.019) were associated with a reduced probability of clinical improvement. CONCLUSION: Certain patients with intermittent claudication show substantial clinical improvement with conservative medical therapy, despite any lack of hemodynamic improvement. Given the low number of patients with clinical deterioration in the short term, primarily conservative therapy should be the preferred initial option for most claudicants. PMID- 14760594 TI - Primary varicose veins: altered transcription of VEGF and its receptors (KDR, flt 1, soluble flt-1) with sapheno-femoral junction incompetence. AB - OBJECTIVE: Loss of regulation of vascular endothelial growth factor (VEGF) production and action disturbs vascular homeostasis leading to pathology. Primary varicose veins (VVs) demonstrate aberrant production/release of VEGF. Our aim was to examine transcription of genes for VEGF (VEGF(121)/VEGF(165)) and its receptors (KDR, flt-1, s.flt-1) in VVs, in relation to underlying venous incompetence. MATERIALS AND METHODS: Samples of varicose (n=83, 18 patients) or normal (n=14, five subjects) great saphenous vein were divided into segments, determined by anatomical position from the sapheno-femoral junction (SFJ). SFJ and segmental incompetence were determined from duplex scans. Gene transcripts were amplified by RT-PCR, analysed by scanning densitometry, and the levels of transcription determined by ratio to control gene GADPH-3 (GAP-3). RESULTS: VEGF(121)/(165), KDR and flt-1 transcription was elevated in VVs overall (p<0.001), and in VVs with an incompetent SFJ (p<0.001), but not when the SFJ was functional; s.flt-1 was unaltered. Notably, gene transcription was unaffected by segmental position, or incompetence. Position below the SFJ correlated with increased transcription of s.flt-1 when the SFJ was incompetent (p<0.04), and s.flt-1 and VEGF(121) when the segment was incompetent (p<0.03). CONCLUSIONS: SFJ incompetence is associated with altered transcription of VEGF and its receptors reflecting an aetiological mechanism or later stage of disease development. Altered VEGF(121) and s.flt-1 transcription may be an early event in varicogenesis. PMID- 14760595 TI - Recurrent varicose veins: incidence, risk factors and groin anatomy. AB - OBJECTIVES: To investigate the recurrence rate after sapheno-femoral junction (SFJ) ligation and great saphenous vein (GSV) stripping for varicose veins (VV), to evaluate risk factors for recurrence and to classify the anatomy of the recurrence in the groin. Design. Clinical follow-up study. METHODS: Eighty-nine consecutive patients with 100 operated legs were re-examined clinically and with duplex after 6-10 years. Fourteen groins were re-explored, 13 after varicography. The anatomy in the groin was classified according to the Edinburgh system. The original medical records were examined to check for risk factors which could lead to a recurrence. RESULTS: Fifty-seven legs had incompetent veins in the groin according to duplex. In 54 of them, it was possible to define whether the incompetent veins emanated from the former SFJ. Varicography and operative findings correlated well to duplex. The main difficulty was to distinguish neovascularization from residual branches. No significant risk factor for recurrence was found in the medical records. CONCLUSIONS: Recurrence of VV after SFJ ligation is common irrespective of perioperative difficulties or the surgeon's experience. The anatomy of recurrence in the groin is difficult to classify according to the Edinburgh system mainly because neovascularization is difficult to verify. PMID- 14760596 TI - Recurrent varicose veins after surgery: a new appraisal of a common and complex problem in vascular surgery. AB - OBJECTIVE: To assess the true incidence, the reflux patterns and the mechanisms responsible for recurrent varicose vein disease according to current definitions and guidelines. PATIENTS AND METHODS: Ninety-three patients (69 female, 24 male, mean age: 48 years) were prospectively evaluated pre- and postoperatively (1 month and 5 years), using clinical and colour duplex examination of both lower limbs. The CEAP score and its modification for recurrence (REVAS) were used for classification. RESULTS: In 113 operated lower limbs, 28 (25%) were found to have a recurrence, 20 of which were symptomatic (20/28, 72%). However, in this group, the mean severity score decreased significantly from 6.5 (SD 3.1) to 5.2 (SD 2.8) (p<0.001, paired t-test). The correlation between the type and cause of recurrence revealed: (1) true recurrent varices in eight limbs (8/28, 29%), primarily caused by neovascularisation, (2) new varicose veins as a consequence of disease progression in seven limbs (7/28, 25%), (3) residual veins in three limbs (3/28, 11%) mainly due to tactical errors (e.g. failure to strip the GSV), (4) complex patterns in 10 limbs (10/28, 36%). In the limbs with recurrence, 42 sources of venous reflux were identified: (1) 19 new sites of venous reflux were due to disease progression (15% of the operated limbs), (2) 13 were caused by neovascularisation (11.5% of the operated limbs), (3) six resulted from tactical failures (5.3% of the operated limbs) and (4) four were due to technical failures (3.5% of the operated limbs). CONCLUSIONS: This study shows that the recurrence of varicose veins after surgery is not uncommon. However, the clinical condition of most affected limbs remains improved. Progression of the disease and neovascularisation are responsible for more than half of the recurrences. Rigorous evaluation of patients and assiduous surgical technique might reduce recurrence due to technical and tactical failures. PMID- 14760597 TI - The efficacy of robot-assisted versus conventional laparoscopic vascular anastomoses in an experimental model. AB - BACKGROUND: Robot-assisted surgery is thought to facilitate complex laparoscopic movements, enhancing advanced laparoscopic procedures. OBJECTIVE: To evaluate the benefit of robotic assistance for laparoscopic vascular surgery. DESIGN: Experimental study using prosthetic conduits in a laparoscopic training box. METHODS: Two surgeons each performed 40 laparoscopic vascular anastomoses alternating with and without robotic assistance. A Zeus-Aesop surgical Robotic system trade mark with 3-D visualisation was used. Each surgeon made 40 anastomoses in total, using different prostheses (5 mm PTFE and 16 mm Dacron) and suture material (Prolene and PTFE). A time-action analysis was performed to evaluate surgical performance. Primary efficacy parameters were quality and leakage of the anastomosis, total time and total number of actions. RESULTS: Equal quality scores and anastomotic leakage were achieved with both techniques. Robotic assistance resulted in significant longer suture and knot tying time and significant more actions were needed compared to the manual laparoscopic procedures. Significant more failures occurred during the robot-assisted procedures. CONCLUSION: In this study, robotic (Zeus-Aesop) assistance did not improve the laparoscopic performance of the surgeon whilst making vascular anastomoses. PMID- 14760598 TI - Abdominal aortic aneurysm repair in patients with chronic renal disease. AB - OBJECTIVES: To assess the outcome of AAA repair in patients with established renal failure (RF), including patients on dialysis. DESIGN: Retrospective case control study in a teaching hospital. Methods. All patients with established RF undergoing AAA repair were identified during the last eight years. Data was collected from patient notes on operative difficulty, hospital mortality, survival time and future dialysis requirements. For comparison, 28 consecutive patients undergoing AAA repair without RF were studied prospectively. RESULTS: Thirteen RF patients were identified. Three were receiving Continuous Ambulatory Peritoneal Dialysis (CAPD), three were receiving Haemodialysis (HD) and seven had established RF, but were not receiving dialysis. Compared with the control patients, RF patients had a longer total hospital stay (p=0.03, 95% CI for median stay -24.3 to -4.0 days), more postoperative complications (p<0.01, 95% CI 26.4 73.7%) and had an increased in-hospital mortality (p=0.02, 95% CI 4.6-54.3%). Four of the six survivors who were non-dialysis-dependent required long-term dialysis postoperatively. CONCLUSIONS: AAA repair in RF patients is associated with increased postoperative morbidity and mortality. Previously non-dialysis dependent patients have a high risk of subsequent long-term dialysis. PMID- 14760599 TI - The clinical value of the systemic inflammatory response syndrome (SIRS) in abdominal aortic aneurysm repair. AB - OBJECTIVES: The systemic inflammatory response syndrome (SIRS) is common after major surgery. We examine the dynamics of SIRS in AAA patients, and assess the impact of the number of SIRS criteria on patient outcome. DESIGN: Prospective study of 151 consecutive patients with AAA, undergoing repair electively, urgently or with rupture. METHODS: SIRS scores and organ failure scores were recorded prospectively each day for all patients. Outcome measures included length of stay, evidence of organ failure and mortality. RESULTS: The majority of patients developed SIRS postoperatively. Elective patients with a cumulative SIRS score of > or =10 during postoperative days 1-4 were more likely to die, compared to patients with a SIRS score of <10 (p=0.02). The development of SIRS late in the postoperative period (day 5-10) was associated with adverse outcome (death) in elective patients (p=0.01). The actual number of SIRS criteria present did not significantly correlate with either outcome or the incidence of organ failure. CONCLUSIONS: SIRS is common in patients undergoing AAA repair. The SIRS score provides useful information regarding a patient's physiological state. High SIRS scores, and the development of SIRS late in the postoperative period are associated with adverse outcome in elective patients, and can therefore be used as an indicator of potential problems. PMID- 14760600 TI - Lifetime gain related to cost of repair of ruptured abdominal aortic aneurysm in octogenarians. AB - OBJECTIVE: To report cost related to gained life years after repair of ruptured abdominal aortic aneurysms in patients aged 80 or older. DESIGN: A retrospective study based on prospectively registered data. PATIENTS AND METHODS: Fifty-three patients aged 80 or older were operated on for ruptured abdominal aortic aneurysm over a 20-year period from 1983 to 2002. Thirty-one (58%) patients had systolic BT <80 mmHg. Operative mortality (<30 days) and long-term survival were studied. The number of life-years gained from the operations was estimated. Based on diagnose related group (DRG) values, the cost of each gained life-year was calculated. RESULTS: The operative mortality was 47%. Long-term survival of those patients who survived the operation was similar to that of an age and sex matched population. The 53 operations resulted in 145 gained life-years, which leaves a mean survival of 2.7 years of all the patients and 5.2 years of those who survived the operation. The estimated cost per gained life year was euro6817. CONCLUSIONS: The operative mortality of ruptured abdominal aortic aneurysm remains high. The long-term survival of patients who survive the operation is acceptable. The price of each gained life-year is low, as compared to other established treatment modalities. Improved results with endovascular treatment may even decrease the cost per gained life year. PMID- 14760601 TI - Aneurysmal hypertension and its relationship to sac thrombus: a semi-qualitative analysis by experimental fluid mechanics. AB - OBJECTIVES: To ascertain the effect of aneurysm thrombus and luminal diameter on arterial blood pressure within the abdominal aortic aneurysm lumen and at the sac wall. METHODS: A life-like abdominal aortic aneurysm was incorporated in a pulsatile flow unit, using systemic blood pressure settings of 140/100 mmHg and 130/90 mmHg (denoted the high and low settings, respectively). Aneurysm sac pressure was measured in the absence of thrombus within the sac. This was repeated after a thrombus analogue (gelatine) was introduced into the aneurysm model in an asymmetric fashion. Luminal and sac wall pressures were compared to the systemic pressure, and to each other, in both blood pressure settings. Statistical analysis was performed using ANOVA in Minitab 13. RESULTS: In the empty sac, the luminal and sac wall pressures were identical to the systemic pressures at the high and low settings. After introduction of thrombus, pressure was transmitted in a monophasic pulsatile fashion, measuring 166/142/151 mmHg (SP/DP/MP) at the sac wall, while the corresponding intraluminal pressure was 164/136/145 mmHg (p<0.001, high setting). By contrast, in the low setting, these readings were 157/133/141 (sac wall) and 160/128/138 mmHg (lumen; p<0.001). The sac wall pressures were significantly higher than the luminal pressures for both high and low settings (p<0.001). CONCLUSIONS: Thrombus has a significant effect on the intraaneurysmal lumen itself and causes localised hypertension with high intraluminal pressures. The differences between the sac wall/luminal pressures may affect regional aneurysm wall biomechanics, but needs further study. PMID- 14760602 TI - The Vanguard endovascular stent-graft: mid-term results from a single centre. AB - PURPOSE: Despite initial enthusiasm for endovascular aortic repair, few descriptions of longer-term follow-up of any endovascular device have been published. This paper represents the experience of a single centre with the Vanguard device over a 5-year period. METHODS: Fifty-five patients with a median age of 71 years (range 45-87 years) and aneurysm diameter of 59 mm (45-84 mm) received a bifurcated Vanguard stent-graft between December 1995 and July 1999. Follow-up was according to the Eurostar criteria (clinical assessment, plain film radiography and computed tomography) at 1, 3, 6, 12, 18 and 24 months and then annually thereafter. RESULTS: All primary stent deployments were successful. Median duration of surgery was 120 min (70-360 min). Median post-operative stay was 3 days (1-19 days) with a peri-operative mortality of 5.5%. In the follow-up period (median 40 months, range 6-64 months) there was one aneurysm associated death, and 14 deaths due to other causes. There have been three device migrations, 12 occluded graft limbs, four type II endoleaks and nine type III endoleaks. At 48 months, this has resulted in a survival rate of 67%, an endoleak free survival of 81% and intervention free survival of 59% (Kaplan-Meier). CONCLUSION: Medium term results with the Vanguard device appear to be at least equivalent to open repair with regard to morbidity and mortality. Nevertheless, several delayed complications appear to be related to endograft limb distortion. Important lessons have been learnt in relation to the deployment of bifurcated endografts to reduce the incidence of secondary limb related problems. PMID- 14760603 TI - Mid-term results after endovascular repair of abdominal aortic aneurysms: a four year experience. AB - OBJECTIVES: The purpose of this retrospective, single-institution study is to describe a 4-year experience of examining early and late clinical outcomes after endovascular repair of abdominal aortic aneurysm (AAA). MATERIALS AND METHODS: Between October 1998 and January 2003, 455 patients were submitted for AAA treatment, of whom 269 underwent open repair and 186 were treated with an endovascular procedure. All endovascular-treated patients underwent preoperative arteriography, contrast enhanced CT scanning or spiral-CT to define the morphological characteristics of the aneurysm, including precise diameter and length measurements. RESULTS: Technical success was achieved in 182 (98%) of the endovascular procedures, as intraoperative conversions to open repair and/or aborted procedures occurred in four patients. The perioperative (30-day) mortality rate was 1% (two patients). During the follow-up period (9-60 months) CT, duplex ultrasound scanning and plain abdominal X-ray evaluation were performed at 3, 6, 12 months, and annually thereafter. Type I endoleak occurred in 12 patients (6.6%), required a further endovascular procedure (11) or late conversion to open repair (1). Type II endoleak occurred in five patients (3%). CONCLUSIONS: In our clinical experience the endovascular repair of AAA is a safe and effective technique with good mid-term results in patients at standard and high risk. PMID- 14760604 TI - Early experience with the minimal extracorporeal circulation system (MECC) during thoracoabdominal aortic aneurysm repair. AB - AIM: The purpose of this report is to describe our early experience with a minimal extracorporeal circulation system (MECC), a compact closed heparin coated system consisting of a centrifugal pump and a membrane oxygenator, during thoracoabdominal aortic aneurysm (TAAA) repair. METHODS: Between January and December 2002 the MECC system was employed in seven consecutive patients (four TAAA type II, two TAAA type I and one TAAA type III according to the Crawford classification). In all patients distal aortic, selective renal and visceral perfusion was performed with this compact closed heparin coated system consisting of a centrifugal pump and a membrane oxygenator. RESULTS: The MECC system was used in all cases with no technical malfunctions. Six out seven patients were discharged from the unit. One patient developed paraplegia after TAAA repair and died on the third post-operative day from multi-organ failure. In this case the total spinal ischaemic time was 120 min and the distal aortic perfusion pressure was <50 mmHg. No cardiac, cerebral, renal, hepatic or bleeding complications were recorded in the remaining six patients. CONCLUSION: Our early experience with MECC during TAAA repair showed that it is feasible for distal aortic spinal and visceral selective perfusion. Further large clinical trials are required to determine the efficacy of this technique. PMID- 14760605 TI - Popliteal artery entrapment associated with cannabis arteritis. AB - OBJECTIVE: To report popliteal artery entrapment in a patient with distal necrosis and cannabis-related arteritis, two rare or exceptional disorders never described in association. To conduct a targeted review and especially to seek information on the clinical presentation with characteristics specific to each disorder so as to hasten the diagnosis and choose appropriate management. MATERIAL AND METHODS: A 19-year-old man who presented with plantar claudication associated with necrosis in a toe underwent diagnostic arteriography and surgery for popliteal artery entrapment type III. RESULTS: Surgical clearance resolved the popliteal artery entrapment but left the clinical symptoms unchanged. Closer questioning disclosed a history of cannabis consumption and intravenous vasodilatory therapy was started. After the 21-day course of vasodilator agents the pain disappeared and the toe necrosis regressed. The patient stopped taking cannabis and had no signs of recurrence. CONCLUSION: Whereas a popliteal artery entrapment, albeit a rare event, is well described and responds to standardized treatment, popliteal artery entrapment associated with cannabis-induced arteritis is an exceptional event that could confuse management. Because young people-the age group mainly at risk for popliteal artery entrapment-increasingly use cannabis, cannabis arteritis could become a more frequent event associated with other arterial disorders that may confuse the diagnosis and complicate management. Our experience in a young patient suggests that coexisting popliteal artery entrapment and distal necrosis in a young patient should raise a strong suspicion of an associated vascular disorder possibly related to cannabis consumption. Intravenous vasodilatation treatment is successful provided that cannabis use is discontinued. PMID- 14760606 TI - Treatment of type II endoleaks after endovascular repair of abdominal aortic aneurysms: translumbar puncture and injection of thrombin into the aneurysm sac. AB - OBJECTIVES: The aim of this article is to report our experience in the use of a new technique for the treatment of type II endoleaks which appear after the endovascular treatment of abdominal aortic aneurysms. MATERIALS AND METHODS: In three patients with secondary type II endoleaks, we performed a translumbar puncture, introducing a 22-Gauge needle into the aneurysm sac under CT guidance. Once intrasac pressure had been registered, 1000U (2 ml) of human thrombin were slowly injected into the sac. RESULTS: Complete sealing of the endoleak was achieved in all three patients, confirmed by the lack of contrast filling of the sac in the CT scans performed 5 min and 24 h after the procedure. Initial intrasac pressure was equal to systolic arterial pressure in the three patients. After the procedure, the pressure decreased by 30-40 mmHg. There were no complications during the procedure, which lasted 45-90 min. No endoleak recurrence has been observed in any of the three cases 6 months later. CONCLUSIONS: We present an alternative method of treating type II endoleaks, which could become the treatment of choice if and when a wider experience confirms our initial good results. PMID- 14760608 TI - Mass spectrometry in the structural analysis of flavonoids. AB - Flavonoids are very common and widespread secondary plant metabolites. They have a wide range of biological and physiological activities and serve as chemotaxonomic marker compounds. Therefore, they have been extensively investigated both in the past and during recent years. The interest in them is still increasing. In the search for new compounds, and also in quality control, there is a need to have reliable methodology for the analysis of flavonoids. Mass spectrometry can make an invaluable contribution because of its high sensitivity, possibilities of coupling with liquid chromatography and the availability of powerful tandem mass spectrometric techniques. A review of currently available mass spectrometric methodology used in the structure elucidation of flavonoids is presented. Sample preparation, liquid chromatographic/mass spectrometric analysis and tandem mass spectrometric procedures for the characterization of flavonoid aglycones, O-glycosides, C-glycosides and acylated glycosides are considered. PMID- 14760609 TI - Solid-state glycation of beta-lactoglobulin by lactose and galactose: localization of the modified amino acids using mass spectrometric techniques. AB - The Maillard reaction is commonly encountered during food processing or storage, and also in human nutrition, hence there is a need for analytical methodologies to identify and characterize the modified proteins. This paper reports specific methods using mass spectrometric techniques to localize protein modifications induced by lactose and galactose on beta-lactoglobulin (beta-Lg) under solid state glycation conditions. The extent of glycation was first determined by liquid chromatography/electrospray ionization mass spectrometry (LC/ESI-MS). The specific identification of lactose-modified amino acid residues was realized using both NanoESI-MS, NanoESI-MS/MS (neutral loss scanning modes) and matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI TOFMS) (with and without guanidination of lysine residues) on unfractionated digests. The results indicated that, after 8.25 h of incubation, the lysine residues were the main targets of lactose-induced modification. In addition to the 15 lysine residues, Leu1 (NH2 terminal) and the Arg124 were also found to be modified, thus leading to a total of 17 different modified amino acid residues (versus 15 found by LC/ESI-MS measurement). In a second set of experiments, different strategies consisting of constant neutral loss and precursor ion scanning were compared to characterize galactose-induced modifications. Owing to the high level of beta-Lg glycation, the combined use of these different strategies appeared to be necessary for determining the galactose-modified sites after 8.25 h of incubation. Thus, among the 22 galactose adducts deduced from the LC/ESI-MS measurement, apart from the N-terminal and classical lysine residues, we also observed a few arginine residues (Arg40, Arg124 and Arg148) that were modified, and also dialkylations on specific lysine residues (Lys47, Lys75). PMID- 14760610 TI - Qualitative study of in vivo melphalan adduct formation in the rat by miniaturized column-switching liquid chromatography coupled with electrospray mass spectrometry. AB - In a general study of DNA adduct formation with melphalan, rats were intravenously injected with a single high dose (10 mg kg(-1)). Adduct formation was studied at the nucleoside level in the target organs liver, bone marrow, kidney and blood with the use of 2D liquid chromatography/tandem mass spectrometry (LC/MS/MS). Adducts of dGuo and dAdo were detected under selected reaction monitoring in liver and bone marrow 10 h after administration of melphalan. In the DNA hydrolysates from kidney and blood a Gua-melphalan adduct was found, although in very low abundance. These first results of the search for in vivo-formed melphalan adducts in the rat showed that our miniaturized LC/MS technique is useful for investigating this type of compound. More experiments will be performed in this area to gather more information about the pharmacokinetics and the quantity of adducts formed. PMID- 14760611 TI - Measurement of the malondialdehyde-2'-deoxyguanosine adduct in human urine by immuno-extraction and liquid chromatography/atmospheric pressure chemical ionization tandem mass spectrometry. AB - The major adduct of malondialdehyde with guanine, M(1)G, was measured in human urine from non-smoking healthy individuals. M1G is a mutagenic DNA lesion and a terminal product of lipid peroxidation in vivo that may be implicated in cancer related to lifestyle and diet. On the basis of a recently developed method for the quantification of M1G as an excreted deoxynucleoside using immuno-extraction purification, chemical NaBH4 reduction and liquid chromatography combined with atmospheric pressure chemical ionization tandem mass spectrometry, we demonstrate that the average 24 h excretion rate of M1G-dR is about 12 +/- 3.8 fmol kg(-1) (n = 5). PMID- 14760612 TI - Effects of ion-pairing reagents on the electrospray signal suppression of sulphonated dyes and intermediates. AB - High-performance liquid chromatography/mass spectrometry (HPLC/MS) analysis of anionic species such as sulphonic acid dyes and intermediates requires volatile ion-pairing mobile phase additives. Six di- and trialkylammonium acetates were compared with tetraalkylammonium salts and ammonium acetate in the concentration range 0-20 mmol l(-1) as mobile phase additives for HPLC/MS of polysulphonated compounds. The effects of the structure and concentration of the ion-pairing reagents on the electrospray response of mono-, di- and tetrasulphonic aromatic acids and acid dyes were studied in detail. Further, five different mass analysers and instrument geometries were compared. A higher signal decrease is observed with linear geometry instruments in comparison to orthogonal or even Z spray geometry mass spectrometers. The concentration of mobile phase additives has a significant influence on the abundance ratios of multiply charged ions in the mass spectra of polysulphonated compounds. The competing ions of sulphonic acids may also cause significant signal suppression. PMID- 14760613 TI - Identification of chlorinated fatty acids in fish by gas chromatography/mass spectrometry with negative ion chemical ionization of pentafluorobenzyl esters. AB - This paper reports the development of a technique for identifying and confirming chlorinated fatty acids previously detected in fish by gas chromatography (GC) with halogen-specific detection (XSD). Fatty acid methyl esters (FAMEs) including chlorinated FAMEs within fractions of reversed-phase high-performance liquid chromatography of transesterified fish extracts were derivatized to pentafluorobenzyl esters, which were subjected to GC/mass spectrometry (MS) with negative ion chemical ionization (NICI). Pentafluorobenzyl esters displayed reasonably good GC characteristics, a very high ionization efficiency and a low degree of fragmentation. Chloride ion chromatograms extracted at m/z 35 and 37 from full scans were utilized for locating traces of chlorinated unknowns in the GC elution profile so that their mass spectra could be readily displayed. Significant ions displayed in the mass spectrum scanned in a narrow range of retention time where a chlorinated unknown was located were evaluated using ion chromatograms extracted at the m/z of these ions. The chromatographic peaks of those ions derived from the analyte were expected to center at that specific retention time, whereas those originating from matrix compounds were not. The isotopic patterns of chlorinated ions were also examined against their theoretical relative abundances. Using this approach, three metabolism-related dichloro fatty acids previously identified by GC/XSD in filet extracts of white sucker sampled downstream from a bleached kraft pulp mill were confirmed: dichlorooctadecanoic, dichlorohexadecanoic and dichlorotetradecanoic acids. In addition, an isomer of dichlorotetradecanoic acid was found in a reference fish sample. As sample preparation is critical in this application, improved conditions for hydrolysis and pentafluorobenzyl esterification are also discussed. PMID- 14760614 TI - Identification and assay of wild-type and modified nucleoside mixtures by turbo ionspray ionization tandem mass spectrometry. AB - A method is presented which allows the identification and assay of a nucleoside in the presence of other analogues and homologues. The method is based on the conventional multiple reaction monitoring approach performed on the [M + H]+ ions of wild-type and modified nucleosides produced by the turbo ionspray ionization method on a triple-quadrupole mass spectrometer. The accuracy of the quantitative determination relies on the evaluation of a response factor rho, which takes into account the kinetics of dissociation of the parent ions into the protonated [B + 2H]+ nucleobase ions. The evaluation of the absolute concentration of each analyte in the examined mixture does not require any previous chromatographic separation. PMID- 14760615 TI - Structural studies of the allelic wheat glutenin subunits 1Bx7 and 1Bx20 by matrix-assisted laser desorption/ionization mass spectrometry and high performance liquid chromatography/electrospray ionization mass spectrometry. AB - Structural studies of the high molecular mass (HMM) glutenin subunits 1Bx7 (from cvs Hereward and Galatea) and 1Bx20 (from cv. Bidi17) of bread wheat were conducted using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) and reversed-phase high-performance liquid chromatography/electrospray ionization mass spectrometry (RP-HPLC/ESI-MS). For all three proteins, MALDI-TOFMS analysis showed that the isolated fractions contained a second component with a mass about 650 Da lower than the major component. The testing and correction of the gene-derived amino acid sequences of the three proteins were performed by direct MALDI-TOFMS analysis of their tryptic peptide mixture. Analysis of the digest was performed by recording several MALDI mass spectra of the mixture at low, medium and high mass ranges, optimizing the matrix and the acquisition parameters for each mass range. Complementary data were obtained by RP-HPLC/ESI-MS analysis of the tryptic digest. This resulted in coverage of about 98% of the sequences. In contrast to the gene-derived data, the results obtained demonstrate the insertion of the sequence QPGQGQ between Trp716 and Gln717 of subunit 1Bx7 (cv. Galatea) and a possible single amino acid substitution within the T20 peptide of subunit 1Bx20. Moreover, the mass spectrometric data demonstrated that the lower mass components present in all the fractions correspond to the major components but lack about six amino acid residues, which are probably lost from the protein C-terminus. Finally, the results obtained provide evidence for the lack of glycosylation or other post translational modifications of these subunits. PMID- 14760616 TI - Ion-pair assisted recovery of matrix-assisted laser desorption/ionization mass spectral signals from SDS-containing peptide-protein mixtures. AB - We have developed a simple and effective means of using alkylammonium ion-pairing agents, such as cetyltetramethylammonium bromide, to recover matrix assisted laser desorption/ionization mass spectrometric (MALDI-MS) signals from sodium dodecyl sulfate (SDS)-containing protein and peptide samples. A two-layer method of matrix preparation, with a bottom matrix layer of ion-pairing agents and a top matrix layer of SDS protein samples, is essential for reproducible MALDI mass spectra with good recovery. Both buffer ions and ion-pairing agents have profound effects on signal recovery and can be rapidly and systematically optimized. This practical technique, termed ion-pair assisted recovery (IPAR), is compatible with major SDS-based biotechniques and can be easily incorporated into high-throughput proteomic analysis. PMID- 14760617 TI - Application of a multi-turn time-of-flight mass spectrometer, MULTUM II, to organic compounds ionized by matrix-assisted laser desorption/ionization. AB - The circuit shape of the ion path, or the multi-turn, provides a solution for achieving unrestricted mass resolution from time-of-flight mass analyzers. The potential of a multi-turn type mass spectrometer, the MULTUM II, with a 1.308 m circuit controlled by four toroidal electric sector fields in biological applications was examined. With matrix-assisted laser desorption/ionization, the ion flight of 18 cycles gave a mass resolution of 10,000 for MH+ of protophorphyrin IX. This resolution was correlated with the flight length, and a resolution of 61,000 was achieved for MH+ of angiotensin I after 75 cycles or a 98.75 m total flight. The results demonstrate that the multi-turn mass spectrometer allows not only high resolution but also very high separation of the ions of molecular species from organic compounds. PMID- 14760618 TI - The importance of detector anode size to detector performance. PMID- 14760619 TI - On the conformation-dependent neutralization theory and charging of individual proteins and their non-covalent complexes in the gas phase. PMID- 14760620 TI - Current literature in mass spectrometry. PMID- 14760623 TI - Synthesis of new pyrrolo[2, 3-b]pyridines as a potent inhibitor of tumour necrosis factor alpha. AB - The MAP kinase p38 plays a key role in the biosynthesis of the inflammatory cytokines tumor necrosis factor alpha (TNF-alpha) and 1L-1beta. Accordingly, new pyrrolo[2, 3-]pyridine derivatives 5a-d were prepared from 2-amino-3 cyanopyrroles 3a-d via the intermediate propenylaminopyrroles 4a-d. Then the compounds 5a-d were tested for their ability to inhibit the production of TNF alpha in vivo in rats. The most potent compounds 5a and 5b possess enhanced ability to inhibit the production of TNF-alpha stimulated with bacterial lipopolysaccharide. PMID- 14760622 TI - Amide derivatives of [6-(5-methyl-3-phenylpyrazole-1-yl)-3(2H)-pyridazinone-2 yl]acetic acids as potential analgesic and anti-inflammatory compounds. AB - In this study, amide derivatives of [6-(5-methyl-3-phenyl-pyrazole-1-yl)-3(2H) pyridazinone-2-yl]acetic acid were synthesized and tested for their in vivo analgesic and anti-inflammatory activity by using the p-benzoquinone-induced writhing test and carrageenan-induced hind paw edema model, respectively. The analgesic and anti-inflammatory activity of the compounds 6a, 6d, 6e, 6g, 6h and 6m were more potent than that of aspirin as an analgesic and indomethacin as an anti-inflammatory drug, respectively. The other derivatives generally resulted in comparable activity to reference compounds. Inhibitor activity of the active compounds on cyclooxygenase isoforms was also investigated by using in vitro human whole blood assay and found that these derivatives did not exert their analgesic and anti-inflammatory activities through COX inhibition and other mechanisms might be involved. PMID- 14760624 TI - Styrylquinazolines: a new class of inhibitors on prostaglandin E2 production in lipopolysaccharide-activated macrophage cells. AB - A series of styrylquinazoline derivatives (2a-k) were prepared and evaluated for their inhibiton of prostaglandin E(2) (PGE(2)) production by cyclooxygenase-2 (COX-2). The latter was induced by lipopolysaccharide-stimulated macrophage cells RAW264.7. 3', 4'-Dihydroxylated styrylquinazolines (2a-c), 3'-hydroxylated styrylquinazolines (2h, 2i), and 3'-acetoxy-styrylquinazolines (2j, 2k) exhibited good inhibitory effects of PGE(2) production by COX-2 with a range of IC(50) values of 1.19 approximately 3.56 microM. The potencies were comparable or better than that of the representative stilbene resveratrol (IC(50) = 3.07 microM). These results indicate that styrylquinazolines can be considered as potential resveratrol analogues in the modulation of prostaglandin production by COX-2. PMID- 14760625 TI - Synthesis and antihypertensive activity of 1-(2-thiazolyl)-3, 5-disubstituted -2 pyrazolines. AB - Several substituted 3-aryl-1-(4-aryl-2-thiazolyl)-5-(3-pyridyl)-2-pyrazolines were synthesized by reacting substituted 3-aryl-5-(3-pyridyl)-1-thiocarbamoyl-2 pyrazolines with phenacyl bromide in ethanol. The structures of all compounds were confirmed by IR, (1)H-NMR, mass spectral data and elemental analyses. The antihypertensive activity of compounds was examined by the tail-cuff method and compared with clonidine. Compounds 24-28 showed significant antihypertensive activity. PMID- 14760626 TI - Synthesis and structure-activity relationships of 4-cycloalkylamino-1, 2, 4 triazolo[4, 3-a]quinoxalin-1- one derivatives as A1 and A3 adenosine receptor antagonists. AB - In a previous paper we reported the synthesis and binding activity of 4 cycloalkylamino-1, 2, 4-triazolo[4, 3-a]quinoxalin-1-one derivatives, differently substituted on the appended 2-phenyl ring, some of which were potent and selective A(1) adenosine receptor (AR) antagonists. In the present paper several 4-cycloalkylamino-2-phenyl-1, 2, 4-triazolo[4, 3-a]quinoxalin-1-one derivatives (1-11), bearing simple substituents on the benzofused moiety, are reported. The binding data of bovine A(1) and A(2A) and human A(3) AR show that we have obtained highly potent A(1) AR antagonists. In particular, the 4-cyclohexylamino derivatives 1-5 show higher A(1) vs A(2A) selectivity than the parent compound A, which lacks substituents on the benzofused moiety. Moreover, compounds 1-11 display, in general, good A(3) AR affinity. Finally, SAR studies provide some new insights about the steric requirements of the A(3) receptor pocket, which accommodates the benzofused moiety of our 4-amino-triazoloquinoxalin-1-one derivatives. PMID- 14760627 TI - Synthesis of curcumin analogues as potential antioxidant, cancer chemopreventive agents. AB - New series of 3, 5-bis(substituted benzylidene)-4-piperidones, 2, 7 bis(substituted benzylidene)cycloheptanones, 1, 5-bis(substituted phenyl)-1, 4 pentadien-3-ones, 1, 7-bis(substituted phenyl)-1, 6-heptadien-3, 5-diones, 1, 1 bis(substituted cinnamoyl)-cyclopentanes, and 1, 1-bis(substituted cinnamoyl)cyclohexanes have been synthesized and tested for their antioxidant activity. Among the tested compounds, compounds II(4), II(9) II(10), II(11), V(1), and V(4) exhibited higher free radical scavenger activity with % inhibition values of 90.71, 91.24, 96.91, 94.26, 99.23, and 99.85%, respectively. Moreover, compound V(1) is the safest member toward peripheral multinuclear neutrophils (PMNs) with a % viability value of 91%. Detailed synthesis, spectroscopic, and biological data are reported. PMID- 14760629 TI - Electrophoretic variants of cardiac myosin heavy chain-alpha in Sprague Dawley rats. AB - Analysis of cardiac myosin revealed differences in gel electrophoretic migration patterns of the alpha-isoform of myosin heavy chain, but not the beta-isoform, in Sprague Dawley rats. No differences in the migration patterns of the alpha-or beta-isoforms were observed in other rat strains. Three electrophoretic migration patterns of the alpha-isoforms were observed in individual rats: a slower migrating isoform alone (4% of all rats tested), a faster migrating isoform alone (55%), and both isoforms (41%). The isoform expression pattern was identical in all myocardial regions in each rat. Frequency of expression patterns suggests multiple gene sequences for alpha-cardiac myosin heavy chain in Sprague Dawley rats. Sequence analysis of amplified regions of the Sprague Dawley and Brown Norway rat alpha-myosin genes, specifically the 5'-untranslated region, exons 1 3, and associated introns, showed numerous single nucleotide polymorphisms in coding and noncoding regions, including putative regulatory sites in Sprague Dawley rats, but not in Brown Norway rats. All Sprague Dawley rats varied from Brown Norway rats and no heterogeneity was observed in Brown Norway rats. Several deletions and dimorphic positions were also observed. Dimorphic positions were evident on automated sequencing comparisons. The data indicate that at least two alpha-myosin heavy chain isoforms exist in Sprague Dawley rats and these rats exhibit sequence diversity within that portion of the alpha-myosin heavy chain gene reported in this study. PMID- 14760630 TI - Effect of matrix chain length on the electrophoretic mobility of large linear and branched DNA in polymer solutions. AB - We study the effect of matrix chain molecular weight Mw and concentration c on the electrophoretic mobility micro of large linear and star-like, branched DNA in polymer solutions. Polyethylene oxide (PEO) with narrow molecular weight distributions form the main focus of this study. For PEO concentrations ranging from one half the overlap concentration, c*, to 3c*, the effective drag coefficient, zeta is identical with (mu0/mu) - 1, satisfies the following approximate scaling relationship, zeta approximately cMw(0.7). Here, mu0 is the electrophoretic mobility in free solution. While the concentration dependence is consistent with predictions from the transient entanglement coupling (TEC) model, the molecular weight dependence is significantly weaker. Although a similar dependence of mobility on Mw can be predicted when nonentangling collisions are the dominant source of drag, a model based on these collisions alone cannot reproduce the experimental observations. We also find that the architecture of large DNA does not affect either the concentration dependence or molecular weight dependence of the electrophoretic mobility. PMID- 14760631 TI - A very thin coating for capillary zone electrophoresis of proteins based on a tri(ethylene glycol)-terminated alkyltrichlorosilane. AB - We describe the use of a tri(ethylene glycol)-terminated alkyltrichlorosilane to create a very thin, protein-resistant "self-assembled monolayer" coating on the inner surface of a fused-silica capillary. The same compound has been demonstrated previously on flat silica substrates to resist adsorption of many proteins. As a covalently bound capillary coating, it displays good resistance to the adsorption of cationic proteins, providing clean separations of a mixture of lysozyme, cytochrome c, ribonuclease A, and myoglobin for more than 200 consecutive runs. Electroosmotic flow (EOF) was measured as a function of pH; the coated capillary retains significant cathodal EOF, with roughly 50% of the EOF of an uncoated capillary at neutral pH, making this coating promising for applications requiring some EOF. The EOF was reasonably stable, with a 2.9% relative standard deviation during a 24 h period consisting of 72 consecutive separations of cationic proteins. Efficiencies for cationic protein separations were moderate, in the range of 190,000-290,000 theoretical plates per meter. The coating procedure was simple, requiring only a standard cleaning procedure followed by a rinse with the silane reagent at room temperature. No buffer additives are required to maintain the stability of the coating, making it flexible for a range of applications, potentially including capillary electrophoresis-mass spectrometry (CE-MS). PMID- 14760632 TI - Quantitation of mRNA levels of steroid 5alpha-reductase isozymes: A method that combines one-step reverse transcription-polymerase chain reaction and separation by capillary electrophoresis. AB - We developed an accurate, rapid, and modestly labor-intensive method to precisely quantitate mRNA species by one-step reverse transcription-polymerase chain reaction (RT-PCR). This approach combines the high specificity of quantitative competitive PCR with the sensitivity of laser-induced fluorescence capillary electrophoresis (LIF-CE). Both cDNA synthesis and PCR amplification are performed with the same enzyme and site-specific primers, improving the efficiency of cDNA synthesis. The specific target mRNA and a mimic DNA fragment, used as a competitive internal standard, were coamplified in a single reaction in which the same primers are used. The 5' forward primers were end-labeled with 6-carboxy fluorescein (6-FAM). The ratio of fluorescence intensity between amplified products of the target cDNA and the competitive DNA was determined quantitatively after separation by CE and fluorescence analysis. Using this method, we have been able to precisely quantify the mean amount of steroid 5alpha-reductase (5alpha-R) isozyme mRNA levels in ventral prostate of the rat, detecting 10-fold difference for 5alpha-R1 and 50-fold difference for 5alpha-R2, respectively, in comparison with our previously reported two-step method. Because the competitive RT-PCR presented in this paper enables a more efficient quantitative determination of mRNAs, low-level gene expression could be quantified. PMID- 14760633 TI - On-line concentration of neutral and charged species in capillary electrochromatography with a methacrylate-based monolithic stationary phase. AB - A method involving self-concentration, on-column enrichment and field-amplified sample stacking for on-line concentration in capillary electrochromatography with a polymer monolithic column is presented. Since monolithic columns eliminate the frit fabrication and the problems associated with frits, the experimental conditions could be more flexibly adjusted to obtain higher concentration factor in comparison with conventional particulate packed columns. With self concentration effect, the detection sensitivity of benzene and hexylbenzene is improved by a factor of 4 and 8, respectively. With on-column enrichment and ultralong injection, improvement as high as 22,000 times in detection sensitivity of benzoin is achieved. Furthermore, a combination of the three above-mentioned methods yields up to a 24,000-fold improvement in detection sensitivity for caffeine, a charged compound. Parameters affecting the efficiency of on-line concentration are investigated systematically. In addition, equations describing on-line concentration process are deduced. PMID- 14760634 TI - Application of capillary electrophoresis with different sample stacking strategies for the determination of a group of nonsteroidal anti-inflammatory drugs in the low microg x L(-1) concentration range. AB - Several on-column sample preconcentration modes--large-volume sample stacking using the EOF pump (LVSEP), LVSEP with anion-selective exhaustive injection (LVSEP-ASEI) and field-amplified sample injection with sample matrix removal using the electroosmotic flow (EOF) pump (FAEP)--were used to analyze some nonsteroidal anti-inflammatory drugs (NSAIDs) by capillary electrophoresis, and then compared. Methanol was the background electrolyte solvent to suppress the EOF. The effect of the type and length of the solvent plug, and the sample injection time were investigated in FAEP to determine the conditions that provided the best response. LVSEP, LVSEP-ASEI, and FAEP improved the sensitivity of the peak area by 100-, 1200-, and 1800-fold, respectively. The methodology developed, in combination with solid-phase extraction (SPE), was applied to the analysis of water samples. PMID- 14760635 TI - Development of a fully buffered molybdate electrolyte for capillary electrophoresis with indirect detection and its use for analysis of anions in Bayer liquor. AB - A new counterion-buffered molybdate electrolyte was developed and optimized for simultaneous quantitative determination of up to eight anions (chloride, sulphate, oxalate, fluoride, formate, malonate, succinate, and acetate) in Bayer liquor by capillary electrophoresis with indirect detection at 214 nm. The separation parameters were optimized in respect to separation of the critical analyte group fluoride-formate-malonate, with the optimal electrolyte prepared from molybdic trioxide containing 5.0 mmol/L MoO3, 1.3 mmol/L cetyltrimethylammonium bromide (CTAB), and buffered with diethanolamine (DEA) to pH 9.2 (ca. 20 mM DEA). Total length of separation capillary was 80 cm, resulting in run time of under 4 min. The method is suitable for a wide concentration range of the analytes (1-50 mg/L) with linear calibration plots (R2 = 0.9983-0.9999). Relative standard deviations were 0.05%-0.07% for migration times and 0.67%-2.04% for peak areas. The detection limits were in the range of 0.17-0.51 mg/L or 2-10 micromol/L (hydrostatic injection of 30 s of 1000 x diluted sample). Due to its good buffering capacity, the electrolyte exhibited an excellent ruggedness and good tolerance to the alkaline samples. Consequently, Bayer liquor samples could be diluted as little as 100 x which allows more sensitive determination of minor components over previous methods. The method was successfully applied to analysis of Bayer liquor samples with recoveries in the range of 95-105%. PMID- 14760636 TI - Enantioseparation of new nucleoside analogs, related to d4T and acyclovir, by chiral capillary electrophoresis using highly sulfated beta-cyclodextrins. AB - Baseline separation of some new acyclic nucleosides which are potential antiviral agents was achieved using cyclodextrin capillary zone electrophoresis (CD-CZE). A method for the enantiomeric resolution of these compounds and determination of their enantiomeric purity was developed using anionic CDs (highly sulfated-CD or highly S-CD) as chiral selectors and capillaries, which were dynamically coated with polyethylene oxide (PEO). Operational parameters including (i) the nature and concentration of the chiral selectors, (ii) organic modifiers, (iii) temperature, and (iv) applied voltage were investigated. The use of charged CDs provides (i) a supplementary driving force for the compounds in a running buffer and (ii) enantiomeric resolution by inclusion of compounds in the CD cavity. The highly S-CD was found to be the most effective complexing agent and allowed good enantiomeric resolution. The complete resolution of five nucleoside analogs was obtained using 25 mM phosphate buffer, pH 2.5, containing either highly S-alpha CD, S-beta-CD or S-gamma-CD at 30 degrees C with an applied field of 0.30 kV/cm. The apparent association constants of the inclusion complexes were calculated. The enantiomer migration order for the molecules investigated was determined and the detection limit of enantiomeric impurities was found to vary between 0.34 to 3.56 ng.mL(-1) for the first enantiomer. PMID- 14760637 TI - Development and validation method for determination of Paroxetine and its metabolites by nonaqueous capillary electrophoresis in human urine. Experimental design for evaluating the ruggedness of the method. AB - A simple, rapid, and sensitive procedure using nonaqueous capillary electrophoresis (NACE) to measure Paroxetine (one of the mostly used antidepressants for mental diseases treatment) and three metabolites has been developed and validated. Optimum separation of paroxetine and metabolites was obtained on a 57 cm x 75 microm capillary using a nonaqueous buffer system of 9:1 methanol-acetonitrile containing 25 mM ammonium acetate and 1% acetic acid, with temperature and voltage of 25 degrees C and 15 kV, respectively, and hydrodynamic injection. Fluoxetine was used as an internal standard. Good results were obtained for different aspects including stability of the solutions, linearity, accuracy, and precision. Detection limits between 9.3 and 23.1 microg.L(-1) were obtained for paroxetine and its metabolites. A ruggedness test of the method was carried out using the Plackett-Burman fractional factorial model with a matrix of 15 experiments. This method has been used to determine paroxetine and its main metabolite B at clinically relevant levels in human urine. Prior to NACE determination, the samples were purified and enriched by means of an extraction preconcentration step with a preconditioned C18 cartridge and eluting the compounds with methanol. PMID- 14760638 TI - Creatinine determination in serum by capillary electrophoresis. AB - Creatinine in human serum was separated in a fused-silica capillary with H3PO4 (75 mmol/L, pH 2.5) as BGE, followed by UV detection at 200 nm. Serum with methylimidazole added as internal standard was deproteinized with acetonitrile and the supernatant, after dilution with water was injected at pressure mode. Creatinine and methylimidazole were baseline-resolved in 6.5 min. Linearity in the 0-880 micromol/L range gave an r2 > or = 0.998, recovery was 102 +/- 2.8% (n = 6). Enzymatic breakdown with creatininase confirmed that serum does not interfere. The within-day and between-days coefficient of variation (CV) were < or = 2.16 and 2.7%, respectively. The accuracy, determined for lyophilized samples by isotope dilution gas chromatography-mass spectrometry was < or = +/- 2.0%. The results were compared with HPLC for 32 lyophilized samples and on 27 serum pools. Capillary electrophoresis, rapid and inexpensive, seems a promising alternative to high-performance liquid chromatography (HPLC) for creatinine determination in human serum. PMID- 14760639 TI - Capillary zone electrophoresis for determination of carbohydrate-deficient transferrin in human serum. AB - Carbohydrate-deficient transferrin (CDT) is the most specific marker for diagnosis of chronic excessive alcohol consumption and includes the serum transferrin (Tf) isoforms with two or less sialic acid residues (di-, mono-, and asialo-Tf). To monitor serum CDT, we developed a capillary zone electrophoresis (CZE) method based on the dynamic capillary coating with diethylenetriamine (DETA). The separation was performed in a bare fused-silica capillary (50 microm ID, 57 cm in length), applying a voltage of 25 kV and a temperature of 40 degrees C. Using a 100 mmol/L borate buffer, pH 8.4 with 3 mmol/L DETA, the Tf isoforms (asialo- to pentasialo-Tf) were resolved within 16 min. Enzymatic cleavage of sialic acid residues with neuraminidase and immunosubtraction were used to identify CDT isoforms. The relative amount of CDT expressed as area % of disialo Tf isoform related to the area of tetrasialo-Tf in 50 healthy donors (24 males and 26 females; aged 25-50 years) was 3.15 +/- 0.76% (mean +/- SD). The comparison between CDT values obtained by this CZE procedure and the "Axis-Shield %CDT" kit gave r = 0.644, p < 0.001 (n = 290). This easy to use and inexpensive CZE procedure could be an ideal tool to investigate CDT proteins for clinical or forensic purposes. PMID- 14760640 TI - Sodium dodecyl sulfate-capillary gel electrophoresis of polyethylene glycolylated interferon alpha. AB - Sodium dodecyl sulfate-capillary gel electrophoresis (SDS-CGE) using a hydrophilic replaceable polymer network matrix was applied to characterize the polyethylene glycol(PEG)ylated interferon alpha (PEG-IFN). The SDS-CGE method resulted in a clearer resolution in both the PEG-IFN species and the native IFN species. The distribution profile of PEGylation determined by SDS-CGE was consistent with that obtained by SDS-polyacrylamide gel electrophoresis (PAGE) with Coomassie blue or barium iodide staining. The result was also compared using matrix-assisted laser desorption/ionization-time of flight-mass spectrometry. SDS CGE was also useful for monitoring the PEGylation reaction to optimize the reaction conditions, such as reaction molar ratio. This study shows the potential of SDS-CGE as a new method for characterizing the PEGylated proteins with advantages of speed, minimal sample consumption and high resolution. PMID- 14760641 TI - Analysis of amino acids in individual human erythrocytes by capillary electrophoresis with electroporation for intracellular derivatization and laser induced fluorescence detection. AB - A method for monitoring amino acids in single erythrocytes is described. For intracellular derivatization, reagent fluorescein isothiocyanate (FITC) was introduced into living cells by electroporation. For an 8 microm erythrocyte, the analytes were diluted by a factor of only 1.6. After completion of the derivatization reaction, a single cell was injected into the separation capillary tip and lysed there. The derivatized amino acids were separated by capillary electrophoresis, followed by laser-induced fluorescence detection. Nine amino acids were quantitatively determined, with amounts of amino acids ranging from 3.8-32 amol/single cell. PMID- 14760642 TI - Determination of carbohydrates in juices by capillary electrophoresis, high performance liquid chromatography, and matrix-assisted laser desorption/ionization-time of flight-mass spectrometry. AB - The objective of this work was to develop a sample preparation procedure for determination of the carbohydrate profiles in commercial juice samples by three principally different analytical methods: capillary electrophoresis (CE) with indirect detection, high-performance liquid chromatography (HPLC), and matrix assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF MS). The preparation and purification of juice samples prior to analysis is described. The method using Carrez reagents was found to be an efficient preparation tool for all three methods. The addition of Carrez reagents to the samples for mass analysis improved the quality of the mass spectra of oligosaccharides. The amounts of glucose, fructose, and sucrose as major carbohydrates in fruit juices measured by CE using a simple instrument are in good agreement with the HPLC values and the data declared by the producers of the juices. The results from both methods are critically evaluated and their impact for studies of authenticity is discussed. The decrease of sucrose amount during the storage of samples was explained by acid hydrolysis of this disaccharide. PMID- 14760643 TI - Native and sodium dodecyl sulfate-capillary gel electrophoresis of proteins on a single microchip. AB - Simultaneous electrophoresis of both native and Sodium dodecyl sulfate (SDS) proteins was observed on a single microchip within 20 min. The capillary array prevented lateral diffusion of SDS components and avoided cross contamination of native protein samples. The planar sputtered electrode format provided a more uniform distribution of separation voltage into each of the 36 parallel microchannel capillaries than platinum wire electrodes commonly used in conventional electrophoresis. The customized geometry of the stacking capillary machined into the cover plate of the microchip facilitated reproducible sample injection without the requirement for stacking gel. Polyimide served as a mask and facilitated insulation of the anode and cathode to prevent electrode lift off and deterioration during continuous electrophoresis, even at a constant current of 8 mA. Improved protein separation was observed during capillary electrophoresis at lower currents. Ferguson plot analysis confirmed the electrophoretic mobility of native globular proteins in accordance with their charge and size. Corresponding Ferguson plot analysis of SDS-associated proteins on the same chip confirmed separation of marker proteins according to their molecular weight. PMID- 14760644 TI - Explorative data analysis of two-dimensional electrophoresis gels. AB - Methods for classification of two-dimensional (2-DE) electrophoresis gels based on multivariate data analysis are demonstrated. Two-dimensional gels of ten wheat varieties are analyzed and it is demonstrated how to classify the wheat varieties in two qualities and a method for initial screening of gels is presented. First, an approach is demonstrated in which no prior knowledge of the separated proteins is used. Alignment of the gels followed by a simple transformation of data makes it possible to analyze the gels in an automated explorative manner by principal component analysis, to determine if the gels should be further analyzed. A more detailed approach is done by analyzing spot volume lists by principal components analysis and partial least square regression. The use of spot volume data offers a mean to investigate the spot pattern and link the classified protein patterns to distinct spots on the gels for further investigation. The explorative approach in analysis of 2-D gels makes it possible, in a fast and convenient way, to screen many gels in order to determine the protein patterns that form clusters and could be selected for further examination. PMID- 14760645 TI - Two-dimensional protein database of human pancreas. AB - We report here the two-dimensional protein database of human pancreas. The proteins were analyzed by two-dimensional electrophoresis followed by matrix assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF MS). Totally, 302 proteins were identified, of which about 27% were enzymes with a broad range of catalytic activities. Several of these are specifically expressed in pancreas, such as pancreatic amylase, pancreatic stone protein, pancreatitis-associated protein, pancreatic lipase, pancreatic elastase, etc. Structural and cytoskeletal proteins are also strongly represented on the gels. Thus, the pancreatic proteome reflects the organ's function. This work paves the way for further studies on pancreatic protein expression in health and disease, such as diabetes and pancreatic cancer. PMID- 14760646 TI - Biochemical characterization of symbiosome membrane proteins from Medicago truncatula root nodules. AB - The symbiosome membrane represents a specialized plant membrane that forms both a structural and a functional interface between the legume plant and its bacterial counterpart. In this study, the symbiosome membrane protein profile from the model system Medicago truncatula and the corresponding bacterium Sinorhizobium meliloti was examined using two-dimensional electrophoresis and microcapillary high-performance liquid chromatography (HPLC) tandem mass spectrometry. The identities of 51 proteins were obtained and these proteins were categorized into functional classes to indicate biochemical roles. Symbiosome membrane proteins include an H(+)-ATPase, ENOD16, ENOD8, nodulin-25, BiP, HSP70, PDI, multifunctional aquaporin, a putative syntaxin, and other proteins of known and unknown identity and function. The majority of the proteins identified were involved with protein destination and storage. These results allow us to understand better the biochemical composition of the symbiosome membrane and thus provide a basis to hypothesize mechanisms of symbiosome membrane formation and function. PMID- 14760647 TI - Defining the plant disulfide proteome. AB - There is considerable interest in redox regulation and new targets for thioredoxin and glutaredoxin are now being identified. It would be of great benefit to the field to have a list of all possible candidates for redox regulation--that is all disulfide proteins in plant. We developed a simple and very powerful method for identifying proteins with disulfide bonds in vivo. In this method, free thiols in proteins are fully blocked by alkylation, following which disulfide cysteines are converted to sulfhydryl groups by reduction. Finally, proteins with sulfhydryls are isolated by thiol affinity chromatography. Our method is unique in that membrane proteins as well as water-soluble proteins are examined for their disulfide nature. By applying this method to Arabidopsis thaliana we identified 65 putative disulfide proteins, including 20 that had not previously been demonstrated to be regulated by redox state. The newly identified, possibly redox-regulated proteins include: violaxanthin de-epoxidase, two oxygen-evolving enhancer proteins, carbonic anhydrase, photosystem I reaction center subunit N, photosystem I subunit III, S-adenosyl-L-methionine carboxyl methyltransferase, guanylate kinase, and bacterial mutT homolog. Possible functions of disulfide bonding in these proteins are discussed. PMID- 14760651 TI - In search of evolutionary developmental mechanisms: the 30-year gap between 1944 and 1974. AB - The approach I have elected in this retrospective of how I became a student of evo-devo is both biographical and historical, a case study along the lines of Waddington's The Evolution of an Evolutionist ('75), although in my case it is the Evolution of an Evo-devoist. What were the major events that brought me to developmental biology and from there to evo-devo? They were, of course, specific to my generation, to the state of knowledge at the time, and to my own particular circumstances. Although exposed to evolution and embryology as an undergraduate in the 1960s, my PhD and post-PhD research programme lay within developmental biology until the early 1970s. An important formative influence on my studies as an undergraduate was the work of Conrad Hal Waddington (1905-1975), whose writings made me aware of genetic assimilation and gave me an epigenetic approach to my developmental studies. The switch to evo-devo (and my discovery of the existence of the neural crest), I owe to an ASZ (now SICB) symposium held in 1973. PMID- 14760652 TI - Kowalevsky, comparative evolutionary embryology, and the intellectual lineage of evo-devo. AB - Alexander Kowalevsky was one of the most significant 19th century biologists working at the intersection of evolution and embryology. The reinstatement of the Alexander Kowalevsky Medal by the St. Petersburg Society of Naturalists for outstanding contributions to understanding evolutionary relationships in the animal kingdom, evolutionary developmental biology, and comparative zoology is timely now that Evo-devo has emerged as a major research discipline in contemporary biology. Consideration of the intellectual lineage of comparative evolutionary embryology explicitly forces a reconsideration of some current conceptions of the modern emergence of Evo-devo, which has tended to exist in the shadow of experimental embryology throughout the 20th century, especially with respect to the recent success of developmental biology and developmental genetics. In particular we advocate a sharper distinction between the heritage of problems and the heritage of tools for contemporary Evo-devo. We provide brief overviews of the work of N. J. Berrill and D. T. Anderson to illustrate comparative evolutionary embryology in the 20th century, which provides an appropriate contextualization for a conceptual review of our research on the sea urchin genus Heliocidaris over the past two decades. We conclude that keeping research questions rather than experimental capabilities at the forefront of Evo devo may be an antidote to any repeat of the stagnation experienced by the first group of evolutionary developmental biologists over one hundred years ago and acknowledges Kowalevsky's legacy in evolutionary embryology. PMID- 14760653 TI - Trochophora larvae: cell-lineages, ciliary bands, and body regions. 1. Annelida and Mollusca. AB - The trochophora concept and the literature on cleavage patterns and differentiation of ectodermal structures in annelids ("polychaetes") and molluscs are reviewed. The early development shows some variation within both phyla, and the cephalopods have a highly modified development. Nevertheless, there are conspicuous similarities between the early development of the two phyla, related to the highly conserved spiral cleavage pattern. Apical and cerebral ganglia have almost identical origin in the two phyla, and the cell-lineage of the prototroch is identical, except for minor variations between species. The cell-lineage of the metatrochs is almost unknown, but the telotroch of annelids and the "telotroch" of the gastropod Patella originate from the 2d-cell, as does the gastrotroch in the few species which have been studied. The segmented annelid body, i.e. the region behind the peristome, develops through addition of new ectoderm from a ring of 2d-cells just in front of the telotroch. This whole region is thus derived from 2d-cells. Conversely, the mollusc body is covered by descendants of cells from both the C and D quadrants and a growth zone is not apparent. This supports the notion that the molluscs are not segmented like the annelids, and that the repeated structures seen in polyplacophorans and monoplacophorans do not represent a segmentation homologous to that of the annelids. PMID- 14760654 TI - Evo-devo aspects of classical and molecular data in a historical perspective. AB - We discuss the interplay between evolution and development as reflected in data and concepts since about 1800. Darwin and his "continental apostle" Haeckel put the striking similarity between early vertebrate embryos in an evolutionary context. Haeckel's partly illicit generalizations discredited evolutionary thinking among early experimental embryologists who moreover noted riddles incompatible with contemporary concepts of homology and evolution. Relevant solutions were suggested by the more recent concept of ontogenetic networks that embody complex regulatory properties and genes with partly overlapping functions. Molecular data on development increasingly reveal evolutionary opportunism, for instance when a widespread signaling chain involved in primitive immune defense was apparently recruited later on for dorso-ventral axis determination in some evolutionarily advanced insect groups. Recently, Rickettsia-related bacteria colonizing many arthropod species were found to "manipulate" the first steps of host development to the advantage of their own propagation, but by ways that could also promote host speciation. Molecular genetics can now document evolutionary steps in ontogenetic networks. In the fruit fly for instance, the novel bicoid gene has superseded a crucial patterning function within a pre existing network--a case of "molecular caenogenesis." The expression patterns of conserved genes that antagonistically determine dorso-ventral polarity support a literal revolution envisioned almost 200 years ago. This is the dorso-ventral inversion of the body plan in some metazoans--ascribed then to the Articulata, now to the Chordata. The final section posits that the opportunistic character of evolutionary innovations is detrimental to parsimony in development. PMID- 14760655 TI - Rupert Riedl and the re-synthesis of evolutionary and developmental biology: body plans and evolvability. AB - This paper reviews the scientific career of Rupert Riedl and his contributions to evolutionary biology. Rupert Riedl, a native of Vienna, Austria, began his career as a marine biologist who made important contributions to the systematics and anatomy of major invertebrate groups, as well as to marine ecology. When he assumed a professorship at the University of North Carolina in 1968, the predominant thinking in evolutionary biology focused on population genetics, to the virtual exclusion of most of the rest of biology. In this atmosphere Riedl developed his "systems theory" of evolution, which emphasizes the role of functional and developmental integration in limiting and enabling adaptive evolution by natural selection. The main objective of this theory is to account for the observed patterns of morphological evolution, such as the conservation of body plans. In contrast to other "alternative" theories of evolution, Riedl never denied the importance of natural selection as the driving force of evolution, but thought it necessary to contextualize natural selection with the organismal boundary conditions of adaptation. In Riedl's view development is the most important factor besides natural selection in shaping the pattern and processes of morphological evolution. PMID- 14760656 TI - Liver transplantation for hepatocellular carcinoma. PMID- 14760657 TI - Establishing a national tissue bank for surgically harvested cancer tissue. PMID- 14760658 TI - Reflux after oesophagectomy. AB - BACKGROUND: Reflux of gastric and duodenal content after oesophagectomy with gastric conduit reconstruction is a common problem and largely considered an inevitable consequence of surgery. Cervical burning and regurgitation, often more pronounced when supine, can be troublesome and even disabling, interfering substantially with quality of life. The aim of this study was to identify the factors contributing to reflux after oesophagectomy and evaluate measures to prevent or control it. METHODS: A Medline search using the terms 'gastro oesophageal reflux', 'oesophagectomy' and 'antireflux surgery' was conducted. Additional references and search pathways were sourced from the bibliographies of articles located. RESULTS AND CONCLUSION: Reflux after oesophagectomy is a significant problem, with both clinical and pathological consequences. Simple measures to facilitate gastric emptying, such as creating a gastric tube, performing a pyloric drainage procedure and using gastric motility agents, may produce a reduction in symptoms but do not alone control reflux itself. A variety of surgical reconstructions have been used, many of which are either difficult to fashion or not suitable when a radical resection has been performed. A modified fundoplication at the anastomosis seems to be the simplest technique and may be relatively effective in controlling symptoms. The impact of strategies to reduce reflux on quality of life and on pathological sequelae of reflux in the oesophageal remnant remains to be evaluated. PMID- 14760659 TI - Carcinoma showing thymic-like elements--a rare malignancy of the thyroid gland. AB - BACKGROUND: Carcinoma showing thymic-like elements (CASTLE) is a rare tumour of the thyroid of thymic origin. The histological appearance of this tumour may be similar to that of squamous cell carcinoma of the thyroid, but outcome associated with CASTLE is more favourable. METHODS: A systematic literature review was conducted for case reports on CASTLE. A text word search of the Medline database was made with a manual search of the citations from these references. Twenty-two case reports were found. RESULTS: In five patients with tumour-negative lymph nodes no local or distant recurrence was observed. Seventeen patients had unknown or involved lymph nodes. Two patients were excluded from further study: one had no follow-up and one was treated by irradiation only. Of the remaining 15, six had local, three had distant and two had local and distant recurrence. In patients with involved or unknown lymph node status, local recurrence was noted in one of five patients treated by surgery and irradiation, and in seven of ten patients treated by surgery alone. Irradiation or systemic chemotherapy was given to four patients with recurrent tumours, with variable response. CONCLUSION: CASTLE with tumour-negative lymph nodes has a low risk of recurrence and surgery without adjuvant therapy is sufficient. Radiotherapy seems indicated when lymph nodes are tumour positive and can be effective for recurrent tumours. In selected patients surgery for recurrent tumour can improve quality of life and outcome. PMID- 14760660 TI - Randomized clinical trial of virtual reality simulation for laparoscopic skills training. AB - BACKGROUND: This study examined the impact of virtual reality (VR) surgical simulation on improvement of psychomotor skills relevant to the performance of laparoscopic cholecystectomy. METHODS: Sixteen surgical trainees performed a laparoscopic cholecystectomy on patients in the operating room (OR). The participants were then randomized to receive VR training (ten repetitions of all six tasks on the Minimally Invasive Surgical Trainer-Virtual Reality (MIST-VR)) or no training. Subsequently, all subjects performed a further laparoscopic cholecystectomy in the OR. Both operative procedures were recorded on videotape, and assessed by two independent and blinded observers using predefined objective criteria. Time to complete the procedure, error score and economy of movement score were assessed during the laparoscopic procedure in the OR. RESULTS: No differences in baseline variables were found between the two groups. Surgeons who received VR training performed laparoscopic cholecystectomy significantly faster than the control group (P=0.021). Furthermore, those who had VR training showed significantly greater improvement in error (P=0.003) and economy of movement (P=0.003) scores. CONCLUSION: Surgeons who received VR simulator training showed significantly greater improvement in performance in the OR than those in the control group. VR surgical simulation is therefore a valid tool for training of laparoscopic psychomotor skills and could be incorporated into surgical training programmes. PMID- 14760661 TI - Randomized clinical trial comparing an oral carbohydrate beverage with placebo before laparoscopic cholecystectomy. AB - BACKGROUND: Preoperative oral carbohydrate can attenuate postoperative insulin resistance and catabolism, and may have the potential to improve postoperative recovery. There are no data from randomized studies on postoperative clinical outcome after specific surgical procedures. This study evaluated the clinical effects of a preoperative carbohydrate beverage in patients undergoing laparoscopic cholecystectomy. METHODS: Ninety-four patients undergoing laparoscopic cholecystectomy were included in a randomized clinical trial. Patients were randomized to receive 800 ml of an iso-osmolar 12.5 per cent carbohydrate-rich beverage the evening before operation (100 g carbohydrate) and another 400 ml (50 g carbohydrate) 2 h before initiation of anaesthesia, or the same volume of a placebo beverage. The primary endpoint was general well-being the day after operation. Patients were evaluated from 5 days before to 5 days after operation. Daily scores of general well-being, fatigue, appetite and pain, computerized measurements of physical activity and sleep (actigraphy), and subjective sleep quality were recorded. Nausea and vomiting were assessed twice within the first 24 h after surgery. RESULTS: Data from 86 patients were available for statistical analysis, 43 in each treatment group. No significant intergroup differences in general well-being or any other outcome variable were found. CONCLUSION: A preoperative carbohydrate beverage did not improve clinical outcome after laparoscopic cholecystectomy. PMID- 14760662 TI - Experimental comparison of four methods of end-to-side anastomosis with expanded polytetrafluoroethylene. AB - BACKGROUND: Four established techniques of distal end-to-side anastomosis (direct anastomosis, Linton patch, Taylor patch and Miller cuff) were compared to investigate the local distribution of anastomotic intimal hyperplasia. The study aimed to elucidate whether mechanical factors or flow alterations are mainly responsible for the improved patency rates reported for vein cuff interposition techniques in infrainguinal arterial reconstructions using prosthetic graft material. METHODS: Thirty-two expanded polytetrafluoroethylene (ePTFE) femoropopliteal bypass grafts were implanted in 16 sheep using the four anastomotic techniques. After 6 months the grafts were explanted and examined histologically. The local distribution of intimal hyperplasia was determined, particularly for areas of material transition and of high and low shear stress. RESULTS: The mean amount and distribution of intimal hyperplasia were similar for all anastomotic types. Intimal hyperplasia was greatest along all transitions between ePTFE and venous patches, and between ePTFE and recipient artery. It was lower along the transitions between venous patches and artery, and was lowest at the host artery floor. CONCLUSION: Vein interposition did not reduce anastomotic intimal hyperplasia and did not change the distribution patterns of hyperplasia, which were influenced mainly by mechanical factors. The effect of vein interposition is to move areas of maximum intimal hyperplasia away from the small recipient artery up to the more capacious graft-patch anastomosis. PMID- 14760663 TI - Population-based study of carotid endarterectomy in Western Australia. AB - BACKGROUND: Previous studies reported an increase in the rates of operation following the publication of major trials that demonstrated the benefit of carotid endarterectomy in reducing stroke. The aim of this study was to determine whether carotid endarterectomy rates have continued to rise despite the reducing trend in most manifestations of atherosclerotic cardiovascular disease. METHODS: Record linkage was used to select patients who had a carotid endarterectomy during the interval from 1988 to 2001. Incidence rates were age-standardized and trends were examined with Poisson regression. RESULTS: The rate increased by 13.8 per cent per year between 1988 and 1998; however, from 1999 onwards the rate of carotid surgery fell by 15.8 per cent per year. In octogenarians, the rate increased steadily from 0.9 to 5.1 per 100,000 person-years between 1992 and 2000. The proportion of octogenarians also increased significantly from 0.9 per cent in 1988-1990 to 19.5 per cent in 2000-2001 (chi2=60.11, 4 d.f., P<0.001). CONCLUSION: For the first time a recent decline has been observed in the rate of carotid endarterectomy, most likely owing to a combination of the deceasing incidence of atherosclerosis and more widespread use of effective drugs in the treatment of cardiovascular disease. The rate and proportion of operations in patients aged 80 years or older has increased steadily. PMID- 14760664 TI - Late complications after ligation and bypass for popliteal aneurysm. AB - BACKGROUND: Ligation and bypass is standard treatment for popliteal aneurysm. This technique does not abolish collateral circulation to the aneurysm, which may continue to expand and/or rupture. This study assessed whether complete thrombosis of the aneurysm sac occurs after operation and examined the long-term clinical outcome. METHODS: The records of all patients who underwent popliteal aneurysm repair in a university hospital over 10 years were reviewed. Patients who had undergone ligation and bypass were recalled for clinical and ultrasonographic examination to determine the fate of the aneurysm sac. RESULTS: Persistent blood flow in the aneurysm sac was present in 12 of 36 legs a median of 48 months after operation. This was associated with symptomatic enlargement of the aneurysm in six patients. The incidence of sac enlargement was lower in bypassed aneurysms with no intrasac flow on duplex examination. CONCLUSION: Ligation and bypass does not always abolish blood flow in the sac of a popliteal aneurysm. It may be associated with continued expansion and late complications. PMID- 14760665 TI - Electrolytic liver ablation is not associated with evidence of a systemic inflammatory response syndrome. AB - BACKGROUND: Local ablation has been proposed for treatment of liver tumours. Cryoshock, a variant of the systemic inflammatory response syndrome (SIRS), is a potentially fatal complication of cryoablation caused by systemic release of necrotic breakdown products from ablated liver. The proinflammatory cytokines tissue necrosis factor (TNF) alpha and interleukin (IL) 1 are important mediators of this response. This study assessed the risk of SIRS complicating electrolytic liver ablation by measuring circulating levels of inflammatory cytokines, other inflammatory markers and clinical markers of organ function. METHODS: Electrolytic liver ablation was performed in 16 pigs and four pigs served as controls. Platelet count, and serum levels of urea, creatinine, liver enzymes, C reactive protein (CRP), TNF-alpha and IL-1beta were measured before treatment and for 72 h after the procedure. RESULTS: There were significant dose-related increases in CRP and alanine aminotransferase levels with liver electrolysis. There was no significant derangement in renal function or platelet count following ablation. A rise in serum TNF-alpha and IL-1beta levels was not associated with liver electrolysis. CONCLUSION: There was no evidence of organ failure or significantly raised levels of proinflammatory cytokines as a result of liver electrolysis, suggesting that this is a safe procedure for liver ablation. PMID- 14760666 TI - The hydroxyl radical scavenger MCI-186 protects the liver from experimental cold ischaemia-reperfusion injury. AB - BACKGROUND: Oxidative stress contributes to hepatic ischaemia-reperfusion (IR) injury in a biphasic pattern. In addition to direct cytotoxic effects, oxidative stress also initiates the signal transduction processes that promote second-phase liver injury. The present study investigated the effects of the hydroxyl radical scavenger MCI-186 on the biphasic process of hepatic cold IR injury. METHODS: After cold preservation for 16 h, rat livers were reperfused on an isolated liver perfusion system for 120 min with oxygenated Krebs-Henseleit bicarbonate buffer. Perfusate samples were obtained serially, and portal flow rates were also recorded. To determine whether MCI-186 affected cytokine levels that control the second-phase injury, levels of interleukin (IL) 10 and tumour necrosis factor (TNF) alpha were measured in the perfusate. RESULTS: Addition of MCI-186 1 mg/l into the perfusate significantly improved portal flow (P<0.050), hepatic enzyme release into the perfusate (P=0.038), total bile production (P=0.029) and malondialdehyde concentration (P=0.038). Furthermore, treatment with MCI-186 led to a substantial increase in IL-10 release (P=0.032). TNF-alpha levels were not affected. CONCLUSIONS: MCI-186, an agent ready for clinical use, appears to have direct and indirect protective effects against hepatic cold IR injury. PMID- 14760667 TI - Hyperlipidaemia is positively correlated with lymph node metastasis in men with early gastric cancer. AB - BACKGROUND: Although increased dietary fat or cholesterol has been reported to be a risk factor for the development of certain cancers, the effect of serum lipid levels on tumour metastasis is not clearly understood. METHODS: The association between lymph node metastasis and preoperative serum levels of total cholesterol (TC) and triglyceride (TG) as well as various pathological findings for tumours was examined in 353 patients with early gastric cancer who underwent gastrectomy with classical lymphadenectomy. RESULTS: The rate of lymph node metastasis was significantly higher in patients with early gastric cancer who had hypercholesterolaemia (TC 220 mg/dl or greater) or hypertriglyceridaemia (TG 150 mg/dl or greater). The tendency was more prominent in men, and multivariate analysis showed that hypertriglyceridaemia was an independent risk factor for nodal metastasis in men, in addition to pathological invasion to the submucosal layer or to lymphatic vessels. In contrast, neither hypercholesterolaemia nor hypertriglyceridaemia showed a significant association with nodal status in women with early gastric cancer. CONCLUSION: Raised serum lipid levels might favour the development of lymph node metastasis in men with early-stage gastric cancer. In patients with early gastric cancer serum lipid levels should be checked before operation, and the use of minimal local treatments must be considered carefully in male patients with hyperlipidaemia. PMID- 14760668 TI - Use of oesophagogastroscopy to assess the response of oesophageal carcinoma to neoadjuvant therapy. AB - BACKGROUND: Approximately 25 per cent of patients with oesophageal cancer who undergo neoadjuvant chemoradiotherapy have no evidence of tumour in the resected specimen (complete pathological response). Those who do not respond have a poor 5 year survival compared with complete responders, regardless of whether or not they undergo surgery. Selecting for surgery only those who have a response to neoadjuvant therapy has the potential to improve overall survival as well as to rationalize the management of non-responders. This study assessed the accuracy of oesophagogastroscopy in this setting. METHODS: A prospective database of 804 patients undergoing oesophageal resection for carcinoma was reviewed. Endoscopic assessment of the response to neoadjuvant therapy in 100 consecutive patients was compared with the pathological assessment of response. The survival for each level of response was compared. RESULTS: At endoscopy 30 patients were considered to have had a complete response. This was confirmed pathologically in 15 patients. Survival was improved in those with a pathologically confirmed complete response (3-year survival rate 62.4 (s.e. 12.9) per cent) compared with non responders (16.3 (s.e. 6.6) per cent). Those with microscopic residual disease also had an improved 3-year survival rate (46.3 (s.e. 12.2) per cent); however, oesophagogastroscopy failed to identify this subset. CONCLUSION: Oesophagogastroscopy may be useful in the assessment of tumour response to neoadjuvant therapy. However, owing to its poor accuracy patients should not be excluded from further therapeutic intervention on the basis of this assessment alone. PMID- 14760669 TI - Matched study of three methods for palliation of malignant pyloroduodenal obstruction. AB - BACKGROUND: The traditional management of pyloroduodenal obstruction is open gastrojejunostomy (OGJ). More recently laparoscopic gastrojejunostomy (LGJ) and endoscopic stenting (ES) have been introduced. The aim of this study was to evaluate the three approaches to the palliation of malignant pyloroduodenal obstruction. METHODS: All patients who underwent surgery (open and laparoscopic) for malignant pyloroduodenal obstruction at Auckland City Hospital between 1989 and 2002 inclusive were identified from International Classification of Diseases (ICD) 10 codes and from the Otago Surgical Audit Database. Patients who had an endoscopic stent were identified from the Endoscribe database. A review of medical records was conducted and data recorded in a structured pro forma. There were 181 patients with malignant pyloroduodenal obstruction of whom 56 patients had OGJ, 14 had LGJ and 16 had ES. Patients in the LGJ and ES groups were matched with those who underwent OGJ with respect to American Society of Anesthesiologists (ASA) grade (I-V), age (within 10 years) and level of obstruction (pylorus, first part of duodenum D1, D2, D3 and D4). The primary outcomes compared between the groups were time to starting free oral fluids and light diet, length of stay and survival. RESULTS: There were no significant differences in age, sex, ASA grade and level of obstruction between the matched OGJ (n=16), LGJ (n=14) and ES (n=16) groups. There was a significant reduction in time to starting free oral fluids and light diet, and length of stay after the procedure, in the ES group. Patients who underwent surgical palliation of the obstruction had significantly more complications than those who underwent stenting (P=0.016). There were no significant differences in requirement for biliary drainage either before or after the procedure between the three groups. Survival was shortest in the ES group. CONCLUSION: This matched study showed significant advantages for ES compared with OGJ and LGJ in the palliation of malignant pyloroduodenal obstruction. PMID- 14760670 TI - Inadvertent perforation during rectal cancer resection in Norway. AB - BACKGROUND: Inadvertent perforation of the bowel or tumour is a relatively common complication during resection of rectal cancer. The purpose of this study was to examine intraoperative perforation following the introduction of mesorectal excision as a standard surgical technique in Norway. METHODS: This was a prospective national cohort study of 2873 patients undergoing major resection of rectal carcinoma at 54 Norwegian hospitals from November 1993 to December 1999. RESULTS: The overall perforation rate was 8.1 per cent (234 of 2873 patients). In a multivariate analysis, the risk of perforation was significantly greater in patients undergoing abdominoperineal resection (odds ratio (OR) 5.6 (95 per cent confidence interval (c.i.) 3.5 to 8.8)) and in those aged 80 years or more (OR 2.0 (95 per cent c.i. 1.2 to 3.5)). The 5-year local recurrence rate was 28.8 per cent following perforation, compared with 9.9 per cent in patients with no perforation (P<0.001); survival rates were 41.5 and 67.1 per cent respectively (P<0.001). CONCLUSION: The risk of intraoperative perforation was significantly greater in patients with rectal cancer undergoing abdominoperineal resection and in those aged 80 years or more. The high local recurrence rates and reduced survival following perforation call for increased attention to avoid this complication. PMID- 14760671 TI - Percutaneous radiofrequency ablation of pulmonary metastases in patients with colorectal cancer. AB - INTRODUCTION: This study aimed to assess the safety and efficacy of imaging guided percutaneous radiofrequency ablation (RFA) for local control of lung metastases from colorectal cancer (CRC). METHODS: Twenty patients with lung metastases from CRC were treated with a RITA Starburst XL electrode and RITA 1500 generator using temperature control and impedance monitoring. Patients received intravenous sedation and analgesia, or local anaesthetic, and stayed in hospital for at least 24 h after treatment. RFA was assessed with computed tomography (CT) at 1 week and 1 month, and then every 3 months. RESULTS: Forty-four CRC lung metastases in 19 patients were treated successfully at 25 treatment sessions. Five of 19 patients were retreated for new lesions. There were 13 pneumothoraces following the 25 treatments, and six patients required drainage. The median length of follow-up was 730 (range 148-924) days. Six months after treatment CT demonstrated that three lesions had progressed, 25 metastases were stable or smaller, and 11 were no longer visible. At 12 months five metastases had progressed, 11 were smaller or stable, and nine were not visible. CONCLUSION: Percutaneous imaging-guided RFA was associated with modest morbidity. RFA can achieve local control of CRC lung metastases: nine of 25 metastases were not visible on CT at 12 months after treatment. PMID- 14760672 TI - Mechanism of action of botulinum toxin on the internal anal sphincter. AB - BACKGROUND: Botulinum toxin is an effective treatment for anal fissure. Manometric studies support an apparent action of botulinum toxin on the internal anal sphincter (IAS). This aim of this study was to establish the underlying mechanism. METHODS: Porcine IAS strips were suspended in a superfusion organ bath and allowed to equilibrate. Electrical field stimulation (EFS) was applied with parameters that induced nitrergic relaxation followed by noradrenaline-mediated contraction. These responses were compared before and after addition of botulinum toxin. RESULTS: All strips developed myogenic tone, which was slightly increased following the addition of botulinum toxin. EFS-induced nitrergic relaxation was unaffected by toxin treatment. However, EFS-induced contraction was significantly reduced by toxin treatment. 1,1-dimethyl-4-phenylpiperazinium iodide (DMPP), a nicotinic agonist, caused muscle strip contraction, which was blocked by guanethidine, implying the presence of sympathetic ganglia within the IAS. Botulinum toxin significantly attenuated DMPP-induced contraction. CONCLUSION: In the treatment of anal fissure the major effect of botulinum toxin on the IAS is blockade of sympathetic (noradrenaline mediated) neural output. This is probably a postganglionic action, involving a reduction in noradrenaline release at the neuromuscular junction. Botulinum toxin has no significant effect on nitrergic transmission, which is probably not vesicular in nature. PMID- 14760673 TI - Differential effect of nitric oxide synthase inhibition on sigmoid colon longitudinal and circular muscle responses to nicotine and nerve stimulation in vitro. AB - BACKGROUND: Nicotine has been shown to release nitric oxide from nerves in human sigmoid colon. This effect has been used to investigate the innervation and functional relationship of the longitudinal and circular muscle layers. METHODS: Strips of longitudinal and circular muscle were obtained from 19 patients with colorectal cancer. The strips from ten patients were subjected to electrical field stimulation (EFS) in vitro using stimulus parameters for selective stimulation of nerves. The effect of nicotine 1-10 micromol/l on EFS responses was then measured in the presence and absence of a nitric oxide synthase inhibitor, nitro-L-arginine methyl ester (L-NAME) 200 micromol/l. The effect of nicotine on spontaneous activity was investigated in the muscle strips from the other nine patients. RESULTS: Both longitudinal and circular strips responded to EFS with contraction. The time to achieve a peak contractile response (time to peak; TTP) was significantly longer (P<0.001) in circular strips. L-NAME reduced the mean(s.e.m.) TTP in circular muscle from 23.3(2.0) to 17.2(1.5) s (P=0.007) and altered its pattern of response to resemble that of longitudinal muscle. Nicotine 10 micromol/l reduced the contraction to EFS in circular (P<0.001) but not in longitudinal (P=0.347) muscle. The nicotine-induced reduction in circular muscle contraction was blocked by L-NAME 200 micromol/l (P=0.005). CONCLUSION: These findings suggest that nitric oxide release on neural stimulation is greater in circular than in longitudinal muscle. PMID- 14760674 TI - Surgical revision after failure of laparoscopic adjustable gastric banding. AB - BACKGROUND: This paper reports the outcome of surgical revision for complications following laparoscopic placement of an adjustable silicone gastric band (AGB) to treat morbid obesity. METHODS: Seventy-three (19.1 per cent) of 382 patients who underwent laparoscopic AGB placement between January 1996 and March 2001 presented with complications within 6 years after operation. Revision was carried out with the intention of reinstating the functional device in all patients. RESULTS: Successful surgical revision or gradual balloon deflation was performed in 53 patients (29 port-related complications, 14 pouch dilatations, 12 band leakages, three oesophageal dilatations, two symptomatic hernias, one late migration, one intracerebral bleed). Of these patients, 51 (96.2 per cent) had a successful outcome according to the Bariatric Analysis and Reporting Outcome System following significant additional postinterventional weight loss. AGB removal was carried out in 20 patients (13 early or late migrations, five pouch dilatations, three port-related complications, two psychiatric disorders, one band leakage). The final failure rate for complicated AGB procedures was 30.1 per cent. CONCLUSION: AGB placement is associated with a variety of complications. In most cases surgical complications can be treated with minimally invasive surgery, which should allow further weight loss and improvement of quality of life during long-term follow-up. Alternative bariatric procedures should be reserved for patients with poor outcome after surgical revision of the AGB. PMID- 14760675 TI - Serum vascular endothelial growth factor as a tumour marker in soft tissue sarcoma. AB - BACKGROUND: Vascular endothelial growth factor (VEGF) is a potent tumour-produced angiogenic factor. In this study serum levels of VEGF were measured before treatment and during follow-up in patients undergoing primary treatment for suspected soft tissue sarcoma (STS) to assess the value of serum VEGF as a tumour marker. METHODS: Between April 2001 and September 2002, serum VEGF levels were analysed prospectively in 144 patients undergoing primary treatment (surgery, 123; cytotoxic chemotherapy, ten; oral imatinib, eight; radiotherapy, three) for suspected soft tissue sarcoma. Serum VEGF was measured by immunoassay before treatment, in the immediate postoperative interval in patients undergoing surgery, and during follow-up. Serum VEGF concentrations were also measured in 15 healthy volunteers. RESULTS: Median pretreatment serum VEGF levels were significantly raised in patients with grade 2 and grade 3 sarcomas compared with concentrations in patients with benign lesions (413 and 467 versus 233 pg/ml respectively; P=0.007 and P=0.003 respectively). In patients with tumours that had a high level of VEGF expression before treatment, follow-up measurements reflected disease status after treatment. CONCLUSION: Serum VEGF expression correlated with grade in soft tissue sarcoma and reflected response to treatment. PMID- 14760676 TI - Arterioportal shunting as an alternative to microvascular reconstruction after hepatic artery resection. AB - BACKGROUND: Portal vein and hepatic artery resection and reconstruction may be required in radical surgery for biliary cancer. Microvascular reconstruction requires special equipment and training, and may be difficult to accomplish when the arterial stump is small, when there are multiple vessels or when the stump lies deep within the wound. This study examined the feasibility and safety of arterioportal shunting as an alternative to arterial reconstruction. METHODS: Over 30 months, ten patients with biliary cancer (six bile duct and four gallbladder carcinomas) underwent radical surgery with en bloc resection of the hepatic artery and end-to-side arterioportal reconstruction between the common hepatic or gastroduodenal artery and the portal trunk. RESULTS: No patient died. Complications included bile leakage in two patients and liver abscess in one. Routine angiography performed 1 month after surgery revealed shunt occlusion in three patients. Once the existence of hepatopetal arterial collaterals had been confirmed in the remaining patients, the shunt was occluded by coil embolization. CONCLUSION: Arterioportal shunting appears to be a safe alternative to microvascular reconstruction after hepatic artery resection. However, the safety of the procedure and its potential to increase the cure rate require further assessment in a larger series with a longer follow-up. PMID- 14760677 TI - Gastrointestinal stromal tumours (Br J Surg 2003; 90: 1178-1186). PMID- 14760678 TI - Value of endoscopic ultrasonography as a predictor of long-term survival in oesophageal carcinoma (Br J Surg 2003; 90: 1367-1372). PMID- 14760679 TI - Value of endoscopic ultrasonography as a predictor of long-term survival in oesophageal carcinoma (Br J Surg 2003; 90: 1367-1372). PMID- 14760680 TI - Influence of suture material and surgical technique on risk of reoperation after non-mesh open hernia repair (Br J Surg 2003; 90: 1004-1008). PMID- 14760681 TI - Patient to surgeon infections--fact or fiction (Br J Surg 2003; 90: 1315-1316). PMID- 14760682 TI - Patient to surgeon infections--fact or fiction (Br J Surg 2003; 90: 1315-1316). PMID- 14760683 TI - Effect of neoadjuvant chemoradiotherapy on angiogenesis in oesophageal cancer (Br J Surg 2003; 90: 1373-1378). PMID- 14760684 TI - Decline in admission rates for acute appendicitis in England (Br J Surg 2003; 90: 1586-1592). PMID- 14760686 TI - High-level accumulation of a recombinant antibody fragment in the periplasm of Escherichia coli requires a triple-mutant (degP prc spr) host strain. AB - During production of a humanized antibody fragment secreted into the periplasm of Escherichia coli, proteolytic degradation of the light chain was observed. In order to determine which protease(s) were responsible for this degradation, we compared expression of the F(ab')(2) antibody fragment in several E. coli strains carrying mutations in genes encoding periplasmic proteases. Analysis of strains cultured in high cell density fermentations showed that the combination of mutations in degP prc spr was necessary for the cells to produce high levels of the desired recombinant antibody fragment. In order to eliminate the possible effects of mutations in other genes, we constructed E. coli strains with protease mutations in isogenic backgrounds and repeated the studies in high cell density fermentations. Extensive light chain proteolysis persisted in degP strains. However, light chain proteolysis was substantially decreased in prc and prc spr strains, and was further decreased with the introduction of a degP mutation in prc and prc spr mutant strains. These results show that the periplasmic protease Prc (Tsp) is primarily responsible for proteolytic degradation of the light chain during expression of a recombinant antibody fragment in E. coli, and that DegP (HtrA) makes a minor contribution to this degradation as well. The results also show that spr, a suppressor of growth defects in prc strains, is required for a prc mutant to survive throughout high cell density fermentations. PMID- 14760687 TI - Milking microalga Dunaliella salina for beta-carotene production in two-phase bioreactors. AB - A new method was developed for production of beta-carotene from Dunaliella salina. Cells were grown in low light intensity and then transferred to a production bioreactor illuminated at a higher light intensity. It was a two-phase bioreactor consisting of an aqueous and a biocompatible organic phase. Mixing of the cells and extraction were performed by recirculation of the organic phase. Two experiments were performed. In the first experiment, bioreactors were operated at two different solvent recirculation rates of 150 and 200 mL min(-1). The beta-carotene extraction rate increased significantly at the higher recirculation rate, without exerting any influence on cell number and viability. A second experiment was carried out at a recirculation rate (200 mL min(-1)) appropriate for the study of long-term production of beta-carotene. The results show that D. salina at high light intensity remained viable for a long period (>47 days) in the presence of a biocompatible organic phase; however, cell growth was very slow. beta-Carotene could be continuously extracted to the organic phase; the cells continued to produce beta-carotene and the extracted molecules were continuously reproduced. As a result, beta-carotene was continuously removed ("milked") from the cells. beta-Carotene extraction efficiency in this system was >55%, and productivity was 2.45 mg m(-2) day(-1), much higher than that of commercial plants. PMID- 14760688 TI - Mosquito and mammalian cells grown on microcarriers for four-serotype dengue virus production: variations in virus titer, plaque morphology, and replication rate. AB - Dengue (DEN) viruses consisting of four distinct serotypes cause diseases such as dengue fever, dengue hemorrhagic fever, and dengue shock syndrome in humans. Most of the dengue viruses can be effectively propagated in some mosquito and mammalian cell lines. In this study, we applied microcarrier cell culture technology to study two relevant aspects involving dengue virus, one on biotechnology of cell growth and virus production, and the other on virus biology concerning genetic variation of a virus population. We investigated the growth of C6/36 mosquito cells and Vero cells grown on Cytodex 1 microcarriers. High-titer DEN virus production can be achieved in C6/36 and Vero cells infected at low cell inoculation density, in the lag-phase cell stage, and at low multiplicity of infection (MOI). The maximum titers produced for DEN-1, DEN-3, and DEN-4 viruses were approximately 10- to 10,000-fold lower than for DEN-2 virus produced in C6/36 and Vero cells grown on microcarriers. The DEN-2 virus produced in C6/36 cells displayed far more extensive plaque heterogeneity than in Vero cells. Microcarrier C6/36 mosquito cell culture appeared to be the most effective system for four-serotype DEN virus production. Interestingly, some selected variants of DEN virus may outgrow in Vero cells when using a T-flask culture. These results may provide useful information for DEN vaccine development. PMID- 14760689 TI - Removal of Mn(II) ions from aqueous neutral media by manganese-oxidizing fungus in the presence of carbon fiber. AB - A manganese-oxidizing fungus was isolated from a hot spring in Japan. The fungus was increasingly effective at oxidizing Mn(II) ions as the concentration of organic carbon sources in the growth medium was decreased. The fungus oxidized 50 ppm of Mn(II) ions within 160 h in a pH 7.3 medium at 25 degrees C. The presence of carbon fiber shortened the time to 80 h, and promoted steady oxidation. The oxidation products were identified by XPS and XRD to be poorly crystallized and amorphous MnO(2), both with and without the fiber. These results suggest that the fiber participates in kinetically limited oxidation. The fungus was entangled with and clung to the fibers, and the oxidized Mn species accumulated on the fungus. Similarly shaped polyethylene telephthalate fiber did not enhance the oxidation, nor was adhesion of the fungus observed. Although the mechanism is still unknown, the present work shows that removal of Mn from solution through the precipitation and accumulation of Mn-oxides on the fungus in the presence of carbon fiber is a promising improvement for water treatment. PMID- 14760690 TI - Serial flux mapping of Corynebacterium glutamicum during fed-batch L-lysine production using the sensor reactor approach. AB - Using our recently developed sensor reactor approach, lysine-producing, nongrowing Corynebacterium glutamicum MH20-22B cells were subjected to serial (13)C-labeling experiments for flux analysis during the leucine-limited fed-batch production phase in a 300-L bioreactor. Based on two-dimensional (2D) nuclear magnetic resonance (NMR) measurements of (13)C-labeling patterns of cytoplasmic free metabolites, metabolic flux distributions in the central metabolism were successfully determined. Focusing on the highly concentrated metabolite L glutamate, the working hypothesis was validated that the equilibration of labeling patterns in intracellular pools was much faster (up to 9.45 min) than the labeling period (3 h) used in the experiments. Analysis of anaplerotic reactions revealed that highly selective lysine production was accompanied by a significant reduction of decarboxylating reactions from 10 mol% to only 2 mol%, whereas PEP/pyruvate-carboxylating fluxes remained constant at about 40 mol% of consumed glucose. These results support the conclusion that an optimized C. glutamicum L-lysine producer should possess increased PEP carboxylase and/or pyruvate carboxylase activity combined with downregulated, decarboxylating fluxes consuming oxaloacetate/malate. The findings also illustrate the usefulness of the sensor reactor approach in the study of industrial fermentations. PMID- 14760691 TI - Rabies virus production in high vero cell density cultures on macroporous microcarriers. AB - The purpose of the study was to investigate the rabies virus multiplication in Vero cell cultures performed on porous microcarriers, MCs (cellulose-Cytopore and gelatin-Cultispher G), which provide higher available surface area compared with solid (nonporous) MCs (DEAE-Cytodex 1). In a set of experiments performed at the same MC concentration (MCs per milliliter), cell densities regularly obtained in porous MC cultures were comparable, but almost twice as high as those in solid MC cultures. In addition, 41.1 +/- 3.9-, 35.2 +/- 2-, and 19.6 +/- 5.8-fold increases in cell concentration, relative to the initial cell number, along with maximum rabies virus titers of 6.3 +/- 0.3 x 10(4), 5 +/- 0.1 x 10(4), and 4.3 +/ 0.2 x 10(4) FFD(50)/mL were observed in Cytopore, Cultispher G, and Cytodex 1 MC cultures, respectively. When higher concentrations of MCs were employed, lower performances of virus production and MC-cell occupation (cells per MC or cells per square millimeter) were observed. Cell attachment to MCs was shown to be faster for Cytopore MCs and Cytodex 1 MCs than for Cultispher G MCs. Concerning the kinetics of cell multiplication on MCs, exponential cell growth, at similar specific cell growth rates, took place on Cytopore, Cultispher G, and Cytodex 1 MCs. In addition, cell densities as high as 2.1 +/- 0.2 x 10(6) cells/mL on Cytopore MCs, 1.8 +/- 0.1 x 10(6) cells/mL on Cultispher G MCs, and 1 +/- 0.3 x 10(6) cells/mL on Cytodex 1 MCs were regularly obtained in batch cultures. Optical as well as scanning and transmission electron microscopy studies carried out to analyze MC structure, MC cell occupation, and cell permissivity to virus infection demonstrated that there was uniform cell distribution in the external and internal areas of the MCs, suggesting an efficiency of virus synthesis. Our results indicate the usefulness of these supports for rabies virus antigen production, as well as possibilities for further optimization. PMID- 14760692 TI - High-humidity performance of paperboard after treatment with xylanase, endoglucanse, and their combination. AB - For paperboard used to produce packaging, treatment of its fiber constituents with commercial enzymes can improve its compressive strength under cycling and high-humidity conditions. Xylanase that selectively removes pulp xylan (ca. 2% of the pulp by mass) yielded most of the observed beneficial effects, which did not appear related to the packing of the fiber network in the sheet or to the uptake of moisture at high humidity. There was also a marginal increase in the drainability of the pulp slurry, which may increase the rate of papermaking. Although endoglucanase activity also enhances certain pulp properties, there was little benefit in adding it to the xylanase treatment. PMID- 14760693 TI - Restructuring upstream bioprocessing: technological and economical aspects for production of a generic microbial feedstock from wheat. AB - Restructuring and optimization of the conventional fermentation industry for fuel and chemical production is necessary to replace petrochemical production routes. Guided by this concept, a novel biorefinery process has been developed as an alternative to conventional upstream processing routes, leading to the production of a generic fermentation feedstock from wheat. The robustness of Aspergillus awamori as enzyme producer is exploited in a continuous fungal fermentation on whole wheat flour. Vital gluten is extracted as an added-value byproduct by the conventional Martin process from a fraction of the overall wheat used. Enzymatic hydrolysis of gluten-free flour by the enzyme complex produced by A. awamori during fermentation produces a liquid stream rich in glucose (320 g/L). Autolysis of fungal cells produces a micronutrient-rich solution similar to yeast extract (1.6 g/L nitrogen, 0.5 g/L phosphorus). The case-specific combination of these two liquid streams can provide a nutrient-complete fermentation medium for a spectrum of microbial bioconversions for the production of such chemicals as organic acids, amino acids, bioethanol, glycerol, solvents, and microbial biodegradable plastics. Preliminary economic analysis has shown that the operating cost required to produce the feedstock is dependent on the plant capacity, cereal market price, presence and market value of added-value byproducts, labor costs, and mode of processing (batch or continuous). Integration of this process in an existing fermentation plant could lead to the production of a generic feedstock at an operating cost lower than the market price of glucose syrup (90% to 99% glucose) in the EU, provided that the plant capacity exceeds 410 m(3)/day. Further process improvements are also suggested. PMID- 14760694 TI - Effect of doxycycline-regulated calnexin and calreticulin expression on specific thrombopoietin productivity of recombinant Chinese hamster ovary cells. AB - In an attempt to increase the specific thrombopoietin (TPO) productivity (q(TPO)) of recombinant Chinese hamster ovary (rCHO) cells (CHO-TPO), the effect of expression level of calnexin (CNX) and calreticulin (CRT) on q(TPO) was investigated. To control both CNX and CRT expression levels simultaneously, the Tet-Off system was first introduced in CHO-TPO cells, and stable Tet-Off cells (TPO-Tet-Off) were screened by luciferase assay. The doxycycline-regulated CNX and CRT expression system in rCHO cells (TPO-CNX/CRT) was established by cotransfection of CNX and CRT expression vector and pTK-Hyg vector into TPO-Tet Off cells and subsequent screening by Western blot analysis of CNX and CRT. The expression levels of CNX and CRT in TPO-CNX/CRT cells could be tightly controlled by adding different concentrations of doxycycline to a culture medium. Compared with the basal level (2 microg/mL doxycyline), a 2.9-fold increase in CNX expression and a 2.8-fold increase in CRT expression were obtained in the absence of doxycycline. This, in turn, resulted in a 1.9-fold increase in q(TPO), not inhibiting cell growth or changing in vivo biological activity of TPO. Taken together, these results demonstrate that a simultaneous overexpression of CNX and CRT can increase the q(TPO) of rCHO cells. PMID- 14760696 TI - Combinatorial formulation of biocatalyst preparations for increased activity in organic solvents: salt activation of penicillin amidase. AB - A combinatorial experimental technique was used to identify salts and salt mixtures capable of activating penicillin amidase in organic solvents for the transesterification of phenoxyacetate methyl ester with 1-propanol. Penicillin amidase was lyophilized in the presence of various chloride and acetate salts within 96-deep-well plates and catalytic rates measured to determine lead candidates for highly salt-activated preparations. The kinetics of the most active formulations were then further evaluated. These studies revealed that a formulation consisting of 98% (w/w) of a 1:1 KAc:CsCl salt mixture, 1% (w/w) enzyme, and 1% (w/w) potassium phosphate buffer was approximately 35,000-fold more active than the salt-free formulation in hexane, as reflected in values of V(max)/K(m). This extraordinary activation could be extended to more polar solvents, including tert-amyl alcohol, and to formulations with lower total salt contents. A correlation was found between the kosmotropic/chaotropic behavior of the salts (as measured by the Jones-Dole B coefficients) and the observed activation. Strongly chaotropic cations combined with strongly kosmotropic anions yielded the greatest activation, and this is likely due to the influence of the ions on protein-water and protein-salt interactions. PMID- 14760695 TI - Flow calorimetry and dielectric spectroscopy to control the bacterial conversion of toxic substrates into polyhydroxyalcanoates. AB - The microbial conversion of toxic substrates into valuable products in continuous culture requires the equivalent of a tight rope walk between formation of the desired product and intoxication of the microbial catalyst. The condition of the latter is reflected immediately by changes in heat flow rate and beta-dispersion in an electrical RF field. Therefore, these were applied to the example of the continuous growth-associated synthesis of polyhydroxyalcanoates (PHA) from phenol by the bacterial strain Variovorax paradoxus DSM 4065. By controlling the supply of phenol to the chemostat, the rates of degradation, biomass formation, and synthesis of target product, respectively, were increasingly elevated until the onset of poisoning the organisms. The boundary between the maximum rates and the initiation of intoxication coincided with a sudden change in the heat flux. Using this occurrence, it was possible to develop a control strategy and test it successfully for a time period of 80 h. After 40 h the process stabilized at mean values, i.e., at rates of 92% phenol degradation, 100% biomass formation, and 70 75% of PHA formation compared with the situation shortly before poisoning the organisms. Using a moving-average technique to filter the raw dielectric spectroscope data, changes were followed in biomass concentration of approximately 100 mg/L. However, this technique was not sensitive or rapid enough to control the process. PMID- 14760697 TI - Thermal and enzymatic pretreatment of sludge containing phthalate esters prior to mesophilic anaerobic digestion. AB - The present study aimed at investigating the effect of thermal pretreatment of sludge at 70 degrees C on the anaerobic degradation of three commonly found phthalic acid esters (PAE): di-ethyl phthalate (DEP), di-butyl phthalate (DBP), and di-ethylhexyl phthalate (DEHP). Also, the enzymatic treatment at 28 degrees C with a commercial lipase was studied as a way to enhance PAE removal. Pretreatment at 70 degrees C of the sludge containing PAE negatively influenced the anaerobic biodegradability of phthalate esters at 37 degrees C. The observed reduction of PAE biodegradation rates after the thermal pretreatment was found to be proportional to the PAE solubility in water: the higher the solubility, the higher the percentage of the reduction (DEP > DBP > DEHP). PAE were slowly degraded during the pretreatment at 70 degrees C, yet this was probably due to physicochemical reactions than to microbial/biological activity. Therefore, thermal pretreatment of sludge containing PAE should be either avoided or combined with a treatment step focusing on PAE reduction. On the other hand, enzymatic treatment was very efficient in the removal of PAE. The enzymatic degradation of DBP, DEP, and DEHP could be one to two orders of magnitude faster than under normal mesophilic anaerobic conditions. Moreover, the enzymatic treatment resulted in the shortest half-life of DEHP in sludge reported so far. Our study further showed that enzymatic treatment with lipases can be applied to raw sludge and its efficiency does not depend on the solids concentration. PMID- 14760698 TI - Plant proteome analysis. AB - Proteome analysis is becoming a powerful tool in the functional characterization of plants. Due to the availability of vast nucleotide sequence information and based on the progress achieved in sensitive and rapid protein identification by mass spectrometry, proteome approaches open up new perspectives to analyze the complex functions of model plants and crop species at different levels. In this review, an overview is given on proteome studies performed to analyze whole plants or specific tissues with particular emphasis on important physiological processes such as germination. The chapter on subcellular proteome analysis of plants focuses on the progress achieved for plastids and mitochondria but also mentions the difficulties associated with membrane-bound proteins of these organelles. Separate chapters are dedicated to the challenging analysis of woody plants and to the use of proteome approaches to investigate the interaction of plants with pathogens or with symbiotic organisms. Limitations of current techniques and recent conceptual and technological perspectives for plant proteomics are briefly discussed in the final chapter. PMID- 14760699 TI - Application of proteomics in phylogenetic and evolutionary studies. AB - There are few papers that deal specifically with evolutionary studies and proteomics. However, applying proteomics to these studies promises to open new perspectives apropos the construction of phylogenetic trees and the detection of evolutionary changes. Principles and methods of phylogenetic systematics could be used to compare and evaluate proteomes. This would permit the detection and characterization of specific proteins that have evolutionary value in defining monophyly, paraphyly, and polyphyly. PMID- 14760700 TI - Determination of protein markers in human serum: Analysis of protein expression in toxic oil syndrome studies. AB - Toxic oil syndrome (TOS) is a disease that appeared in Spain in 1981. It affected more than 20 000 people and produced over 300 deaths in the first 2 years. In this paper, a prospective study on the differences in gene expression in sera between a control versus a TOS-affected population, both originally exposed to the toxic oil, is presented. Differential protein expression was analyzed by two dimensional electrophoresis (2-DE). Several problems related with serum analysis by 2-DE were addressed in order to improve protein detection in the gel images. Three new commercial systems for albumin depletion were tested to optimize the detection of minor proteins that can be obscured by the presence of a few families of high abundance proteins (albumin, immunoglobulins). Other factors, such as the use of nonionic reductants or the presence of thiourea in the gels, were also tested. From these optimized images, a group of 329 major gel spots was located, matched and compared in serum samples. Thirty-five of these protein spots were found to be under- or overexpressed in TOS patients (> three-fold increase or decrease). Proteins in the differential spots were identified by matrix-assisted laser desorption/ionization-time of flight peptide map fingerprinting and database search. Several haptoglobin isoforms were found to be differentially expressed, showing expression phenotypes that could be related with TOS affection. Haptoglobin phenotypes have been previously reported to have important biological and clinical consequences and have been described as risk factors for several diseases. PMID- 14760701 TI - Identification and characterization of the expression of the translation initiation factor 4A (eIF4A) from Drosophila melanogaster. AB - We have identified the initiation factor 4A (eIF4A) in a two-dimensional protein database of Drosophila wing imaginal discs. eIF4A, a member of the DEAD-box family of RNA helicases, forms the active eIF4F complex that in the presence of eIF4B and eIF4H unwinds the secondary structure of the 5'-UTR of mRNAs during translational initiation. Two-dimensional gel electrophoresis and microsequencing allowed us to purify eIF4A, and generate specific polyclonal antibodies. A combination of immunoblotting and labelling with [(35)S]methionine + [(35)S]cysteine revealed the existence of a single eIF4A isoform encoded by a previously reported gene that maps to chromosome 2L at 26A7-9. Expression of this gene yields two mRNA species, generated by alternative splicing in the 3' untranslated region. The two mRNAs contain the same open reading frame and produce the identical eIF4A protein. No expression was detected of the eIF4A related gene CG7483. We detected eIF4A protein expression in the wing imaginal discs of several Drosophila species, and in haltere, leg 1, leg 2, leg 3, and eye antenna imaginal discs of D. melanogaster. Examination of eIF4A in tumor suppressor mutants showed significantly increased (> 50%) expression in the wing imaginal discs of these larvae. We observed ubiquitous expression of eIF4A mRNA and protein during Drosophila embryogenesis. Yeast two-hybrid analysis demonstrated the in vivo interaction of Drosophila eIF4G with the N-terminal third of eIF4A. PMID- 14760702 TI - Snake venomics: characterization of protein families in Sistrurus barbouri venom by cysteine mapping, N-terminal sequencing, and tandem mass spectrometry analysis. AB - The protein composition of the crude venom of Sistrurus barbouri was analyzed by two-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis. Proteins were separated by reversed phase high-performance liquid chromatography and characterized by N-terminal sequence analysis. The molecular mass and number of cysteine residues of the purified proteins were determined by matrix associated laser desorption/ionization-time of flight mass spectrometry. Selected protein bands were subjected to in-gel tryptic digestion and peptide mass fingerprinting. Analysis of the tandem mass spectrometry spectra of selected doubly-charged peptide ions was done by collision-induced dissociation in a quadrupole-linear ion trap instrument. Our results show that the venom proteome of the pigmy rattlesnake S. barbouri is composed of proteins belonging to a few protein families, which can be structurally characterized by their disulfide bond contents. PMID- 14760703 TI - Proteomic identification of brain proteins that interact with dynein light chain LC8. AB - Cytoplasmic dynein is a large minus end-directed microtubule motor that translocates cargos towards the minus end of microtubules. Light chain 8 of the dynein machinery (LC8) has been reported to interact with a large variety of proteins that possess K/RSTQT or GIQVD motifs in their sequence, hence permitting their transport in a retrograde manner. Yeast two-hybrid analysis has revealed that in brain, LC8 associates directly with several proteins such as neuronal nitric oxide synthase, guanylate kinase domain-associated protein and gephyrin. In this work, we report the identification of over 40 polypeptides, by means of a proteomic approach, that interact with LC8 either directly or indirectly. Many of the neuronal proteins that we identified cluster at the post-synaptic terminal, and some of them such as phosphofructokinase, lactate dehydrogenase or aldolase are directly involved in glutamate metabolism. Other pool of proteins identified displayed the LC8 consensus binding motif. Finally, recombinant LC8 was produced and a library of overlapping dodecapeptides (pepscan) was employed to map the LC8 binding site of some of the proteins that were previously identified using the proteomic approach, hence confirming binding to the consensus binding sites. PMID- 14760704 TI - Application of proteome analysis to seafood authentication. AB - Part of our work aims at studying the modifications that proteins suffer in foods and use them as markers to estimate the origin and history of the product. Proteomics is a powerful approach to do this: comparison of the two-dimensional (2-D) maps of the intact and treated samples would permit to identify marker spots so that in the future it may be possible to estimate the treatment a foodstuff has suffered by examining its 2-D protein map or just the selected markers. This work summarizes some of our previous studies showing the application of proteomics to the (i) identification of species and muscle tissues, (ii) characterization of post-mortem changes in arctic and tropical species, and (iii) study of the effect of some additives during the processing of fish muscle. PMID- 14760705 TI - A proteomic approach to study Salmonella typhi periplasmic proteins altered by a lack of the DsbA thiol: disulfide isomerase. AB - Two-dimensional electrophoresis (2-DE) was used to analyze the pleiotropic effects of a deficiency in DsbA, a periplasmic disulfide-bond oxidoreductase, in Salmonella typhi. With this aim, the dsbA gene was cloned and assayed for activity in a dsbA-null mutant of Escherichia coli. A dsbA/chloramphenicol acetylase construct was then used to disrupt the wild-type gene of S. typhi. The resultant dsbA-null mutant of S. typhi, like the E. coli mutant, exhibited a lack of flagellation and of glucose-1-phosphatase activity. Periplasmic extracts from the parental and mutant strains were analyzed by 2-DE using standard denaturing and nondenaturing conditions. Differences in protein expression were more marked in nondenaturing conditions. Ninety-nine protein spots were analyzed by peptide mass fingerprinting, and 65 spots were identified by searching a S. typhi database. Twenty-five spots were exclusively detected in the wild-type strain, 10 were found only in the mutant strain, and 21 were common to both strains. We observed a lack of DsbA, glucose-1-phosphatase and flagellin in the dsbA-null mutant, which explains two of the observed phenotypes. The AI-2 autoinducer producing protein LuxS, which is involved in quorum-sensing signalling was also absent. PMID- 14760706 TI - Dissecting regulatory networks by means of two-dimensional gel electrophoresis: application to the study of the diauxic shift in the yeast Saccharomyces cerevisiae. AB - Using a proteomic approach based on the two-dimensional (2-D) gel analysis of synthesized proteins, we investigated the involvement of the Snf1 kinase pathway in the regulation of gene expression during the diauxic shift in Saccharomyces cerevisiae. For this purpose, we used a mutant strain deleted for SNF4, the gene coding for the activator subunit of Snf1p. The levels of synthesis of 82 spots were found to be affected by the absence of Snf4p at the diauxic shift. Half of the proteins which exhibit a reduced synthesis in the mutant strain are proteins whose genes are controlled by the transcriptional activator Cat8p, a target of Snf1p. Proteins with an increased level of synthesis in the mutant strain were also observed. Among them are glycolytic enzymes whose synthesis is strongly reduced when wild-type cells enter the diauxic shift. This observation suggests that Snf1p exerts a negative control on the expression of glycolytic genes during the diauxic transition. The results obtained in this study were compiled with those previously obtained by similar proteomic approach with other regulatory factors involved in the diauxic shift. This compilation illustrates how 2-D gel electrophoresis can be used to elucidate the network of regulators participating to complex biological process. PMID- 14760707 TI - Two-dimensional reference map of Candida albicans hyphal forms. AB - We have set up an in vitro model of culture of Candida albicans fungal cells that mimics the physiological conditions found in human beings in order to carry out studies of host-pathogen interaction. Under these conditions, C. albicans undergoes dimorphic transition (yeast-hyphae); this morphological change has been proposed as a virulence factor for this fungus. C. albicans cytoplasmic extract from hyphae cells was analyzed by two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) and a reference map was obtained. Protein identification was carried out by peptide mass fingerprinting or sequence tagging using a matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) or a MALDI-TOF/TOF mass spectrometer. A total of 106 spots, excised from 2-D gels, were analyzed. This resulted in the identification of 43 proteins involved in metabolism, 13 involved in transcription, protein synthesis, and fate, 8 involved in cell rescue, virulence, and defense, and 2 proteins of unknown function. This reference map is an important tool for future studies of protein differential expression after host-pathogen interaction. PMID- 14760708 TI - The defense response of germinating maize embryos against fungal infection: a proteomics approach. AB - Pathogen attack on plants results in numerous host-specific biochemical responses, the activation of some of them being critical for the ability of the plant to withstand disease. We have used high-resolution two-dimensional gel electrophoresis (2-DE) and mass spectrometry to identify proteins that are differentially expressed in response to fungal infection in maize embryos. Differential spots corresponding to induced or repressed proteins were apparent in silver stained 2-DE gels of proteins extracted from sterile and fungal infected germinating embryos. Selected spots were subjected to tryptic digestion followed by identification using matrix-assisted laser desorption/ionization-time of flight-mass spectrometry and nanospray ion-trap tandem mass spectrometry. Among the proteins induced in response to infection are proteins involved in protein synthesis, or in protein folding and stabilization, as well as proteins involved in oxidative stress tolerance. Additionally, the accumulation of specific pathogenesis-related proteins in tissues of the fungal-infected germinating embryos was studied by 2-DE and immunoblotting. PMID- 14760709 TI - Large-scale characterization of integral proteins from Arabidopsis vacuolar membrane by two-dimensional liquid chromatography. AB - We developed a method to characterize different classes of membrane proteins within a single experiment and using simple matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS) analysis. After membrane solubilization with the nondenaturing detergent n-dodecyl-beta-D maltoside, proteins were separated successively by gel filtration and ion exchange chromatography and finally by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). This procedure allowed to characterize 70 proteins from a membrane fraction enriched in plant vacuolar membrane (Arabidopsis), including integral proteins like the V0 complex of the H(+)-ATPase, the H(+) pyrophosphatase or the glutathione S-conjugate ATPase AtMRP1, and peripheral proteins like the subunits of the catalytic V1 complex of the H(+)-ATPase. Approximately 60% of identified proteins were predicted to possess at least two trans-membrane domains. Furthermore, proteins, with molecular masses ranging between 20 and 200 kDa were distributed into two populations with maximum frequencies at pI 5.3 and 8.9. Finally, this procedure appeared to allow the identification of proteins known to be minor in whole-cell extracts like signaling or vesicular trafficking proteins. Almost 50% of the proteins identified were functionally unknown whereas the others confirmed that the plant vacuole is a multipurpose compartment. PMID- 14760710 TI - Identification of specific plant nucleolar phosphoproteins in a functional proteomic analysis. AB - The soluble fraction of nuclear proteins is a functionally significant fraction, since it has been shown that it contains ribonucleoproteins active in nuclear RNA metabolism. The aim of this work was to detect variations associated with cell proliferation, by comparing two-dimensional proteomes obtained from the soluble fractions of onion nuclei isolated from actively proliferating root meristematic cells versus nonmeristematic root cells. In particular, we have studied the physicochemical features of the major nucleolar protein NopA100, a highly phosphorylated, nucleolin-like protein. A total of 384 spots were quantified in meristematic nuclei, while only 209 were detected in nonmeristematic nuclei. The comparison of both proteomes resulted in the determination of specific spots for each proliferative state and those which were common to both cases. Furthermore, among these latter, we could discriminate quantitative differences. Interestingly, well-known nucleolar proteins, such as RNA polymerase I, B23 and the nucleolin-like protein NopA100, were significantly increased in proliferating cells. Western blots with anti-NopA100 antibody demonstrated 26 spots in the meristematic sample. All the spots detected were clustered at 100 kDa and were distributed through an isoelectric point (pI) range of 4.3-6.6. In contrast, only seven spots were found in the extract from nonmeristematic nuclei, and the pI range was shortened to 4.8-6.1. These results indicate that the state of NopA100 phosphorylation correlates with the degree of nucleolar activity, i.e. the protein is more highly phosphorylated in cycling cells. We have also analyzed the bidimensional silver staining of the nucleolar organizing region (Ag-NOR) pattern of the soluble nuclear fraction in order to identify plant cell phosphoproteins that are considered to be markers of proliferation. These experiments demonstrated that NopA100, the onion, nucleolin-like protein, is an Ag-NOR protein. In addition we found that the plant homologue of the vertebrate nucleolar phosphoprotein B23 migrated as two clusters of acidic spots, 43 and 42 kDa respectively in molecular mass. The differences between these features and those described for mammalian cells is discussed. Our results demonstrate that the use of protein fractionation procedures with functional significance and the location of candidate spots by indirect techniques are advantageous, complementary methods to random selection procedures for proteomic studies involving further mass spectrometry analysis. PMID- 14760711 TI - Proteomic analysis of the oxygen-evolving complex of photosystem II under biotec stress: Studies on Nicotiana benthamiana infected with tobamoviruses. AB - We have previously shown that tobamovirus infection induces an inhibition of photosystem II electron transport, disturbing the oxygen-evolving complex (OEC). In the infected plants, the OEC polypeptide pattern was modified when compared to healthy plants, the levels of the PsbP and PsbQ extrinsic proteins being lowered to different extents. In this work we have further investigated by two dimensional polyacrylamide gel electrophoresis (2-DE) the changes on the OEC protein pattern of thylakoid membranes isolated from Nicotiana benthamiana Domin plants infected with the Spanish strain of pepper mild mottle virus. When the thylakoid membranes from healthy plants were analyzed for the presence of PsbO and PsbP proteins by 2-DE (pI range 4-7) and further immunoassayed by using specific-antisera against these two proteins, it was observed that four polypeptides cross-reacted with each antiserum. These data, along with the N terminal amino acid sequence determined for the eight polypeptides, indicate that the N. benthamiana PsbO and PsbP proteins correspond to protein families. In the silver-stained 2-DE gels of thylakoid membranes isolated at different days postinoculation from virus-infected plants, it was observed that the content of PsbP polypeptides decreased dramatically with respect to those of PsbO, during the progress of the infection. Interestingly, there was a differential decrease of the different PsbP proteins, indicative of a distinct regulation of their expression. PMID- 14760712 TI - Applicability of discovery science approach to determine biological effects of mobile phone radiation. AB - We argue that the use of high-throughput screening techniques, although expensive and laborious, is justified and necessary in studies that examine biological effects of mobile phone radiation. The "case of hsp27 protein" presented here suggests that even proteins with only modestly altered (by exposure to mobile phone radiation) expression and activity might have an impact on cell physiology. However, this short communication does not attempt to present the full scientific evidence that is far too large to be presented in a single article and that is being prepared for publication in three separate research articles. Examples of the experimental evidence presented here were designed to show the flow of experimental process demonstrating that the use of high-throughput screening techniques might help in rapid identification of the responding proteins. This, in turn, can help in speeding up of the process of determining whether these changes might affect human health.* PMID- 14760713 TI - Isolation of circulating human monocytes with high purity for proteomic analysis. AB - We describe a simple method for isolation of human blood monocytes with the high purity (95-98%) required for proteomic analysis, which avoids contamination by other blood cells (platelets and lymphocytes) and the most abundant plasma proteins (albumin and immunoglobulins). Blood monocytes were purified by gradient centrifugation followed by positive selection with specific monoclonal antibodies coupled to paramagnetic beads. The elution conditions of the positive selection step were modified to avoid contamination with albumin. This method is compatible with flow cytometry which was used to assess the purity of the cell population. From 28 mL of blood, 10(7) monocytes with > 96% purity are routinely obtained. From the isolated monocytes 200-250 microg of protein could be recovered. The whole method can be performed in three hours. Similar results were obtained using a negative selection step but with lower purity (92%), increased cost and longer time. After solubilization of monocytes, the proteins were analyzed by two dimensional gel electrophoresis (2-DE) in the 3-10, 4-7, 6-9 and 6-11 pH range. DNA was the main contaminant that interfered with the 2-DE and it was removed by treatment with DNAse. Image analysis of gels allowed the reproducible detection and quantification of 1500 spots in the 4-7 pH range and more than 2000 spots in total by combining (overlapping) 2-D gels in the 4-7, 6-9 and 6-11 pH range. This method is useful for clinical studies of monocytes from a large number of patients due to its rapidity and reproducibility, which permits comparative analysis of normal versus pathological samples and which allows follow up of the expressed proteins of monocytes from each patient. PMID- 14760714 TI - Improved resolution of the human adipose tissue proteome at alkaline and wide range pH by the addition of hydroxyethyl disulfide. AB - Polycystic ovary syndrome is a very common disease affecting 6.5% of women. It is frequently associated with obesity with the link between the two disorders being insulin resistance. From morbidly obese women submitted to surgical treatment of obesity, we obtain intra-abdominal adipose tissue samples in order to compare protein patterns between women with and without polycystic ovary syndrome by two dimensional gel electrophoresis (2-DE) analysis. Adipose tissue is a complex material extremely rich in lipids. To improve protein solubilization a lysis buffer containing 8.4 M urea, 2.4 M thiourea, 5% 3-((3-cholamidoproyl)dimethyl amino)-1-propanesulfonate and 50 mM dithiothreitol was used and samples were centrifuged to remove fat. Hydroxyethyl disulfide was added to increase resolution in the alkaline region (Olsson, I., Larsson, K., Palmgren, R., Bjellqvist, B., Proteomics 2002, 2, 1630-1632) and it also improved resolution in a wide pH range (3-10). Our work shows for the first time 2-DE maps of human adipose tissue and identifies some of the proteins by mass spectrometry. This information will aid studies on metabolic diseases such as polycystic ovary syndrome, obesity, hypertension and type 2 diabetes. PMID- 14760715 TI - Proteomic analysis of lung biopsies: Differential protein expression profile between peritumoral and tumoral tissue. AB - The cellular proteome shows a dynamic profile and is subjected to changes in response to various stimuli and disease progression. Lung cancer remains the leading cause of cancer death in industrialized countries. In an attempt to find new disease markers, patients suffering from lung carcinoma have been selected to achieve differential protein expression patterns between normal and neoplasic tissue. After two-dimensional electrophoresis, the spots of interest were digested and identified by matrix-assisted laser desorption/ionization (MALDI) peptide mass fingerprinting. This information will provide a more comprehensive understanding of the disease progression and might constitute a method to complement histopathological diagnosis. PMID- 14760716 TI - Systematic screening of reactive cysteine proteomes. AB - Redox signalling constitutes a topic within the field of cellular signal transduction which is attracting increasing interest. A major challenge is to identify the components of redox signalling pathways. Proteins containing cysteines that may reversibly form disulphides are principal candidates as transmitters of redox signals. Thioredoxins are small proteins containing a highly reactive dithiol. Here we present a simple procedure to isolate and separate proteins that contain redox active cysteines using a site-directed, histidine-tagged mutant of thioredoxin, which forms stable mixed disulphides with its targets. PMID- 14760717 TI - A technical trick for studying proteomics in parallel to transcriptomics in symbiotic root-fungus interactions. AB - We have developed a protocol in which proteins and mRNA can be analyzed from single root samples. This experimental design was validated in arbuscular mycorrhiza by comparing the proteins profiles obtained with those from a classical protein extraction process. It is a step forward to make simultaneous proteome and transcriptiome profiling possible. PMID- 14760718 TI - Screening for N-glycosylated proteins by liquid chromatography mass spectrometry. AB - In the last few years mass spectrometry has become the method of choice for characterization of post-translationally modified proteins. Whereas most protein chemical modifications are binary in the sense that only one change can be associated with a given residue, many different oligosaccharides can be attached to a glycosylation site residue. The detailed characterization of glycoproteins in complex biological samples is extremely challenging. However, information on N glycosylation can be gained at an intermediary level. Here we demonstrate a procedure for mapping N-glycosylation sites in complex mixtures by reducing sample complexity and enriching glycoprotein content. Glycosylated proteins are selected by an initial lectin chromatography step and digested with endoproteinase Lys-C. Glycosylated peptides are then selected from the digest mixture by a second lectin chromatography step. The glycan components are removed with N-glycosidase F and the peptides digested with trypsin before analysis by on line reversed-phase liquid chromatography mass spectrometry. Using two different lectins, concanavalin A and wheat germ agglutinin, this procedure was applied to human serum and a total of 86 N-glycosylation sites in 77 proteins were identified. PMID- 14760719 TI - A strategy to improve peptide mass fingerprinting matches through the optimization of matrix-assisted laser desorption/ionization matrix selection and formulation. AB - Peptide mass fingerprinting (PMF) is a powerful technique in which experimentally measured m/z values of peptides that result from a protein digest form the basis for a characteristic fingerprint of the intact protein. Due to its propensity to generate singly-charged ions, along with its relative insensitivity to salts and buffers, matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) is the MS method of choice for PMF. The qualitative features of a MALDI-MS mass spectrum can be selectively tuned by varying the matrix and the solvent system used to prepare the matrix. The selective tuning of MALDI-MS mass spectra in order to optimize PMF results is addressed in this paper. Carbonic anhydrase, hemoglobin alpha- and beta-chain, and myoglobin were digested with trypsin, and the digest was analyzed with MALDI-MS. 2,5 Dihydroxybenzoic acid (2,5-DHB) and alpha-cyano-4-hydroxycinnamic acid were prepared, using five different solvent systems: (A) 99% acetone; (B) 50% acetonitrile (ACN), 0.1% trifluoroacetic acid (TFA); (C) 75% ACN, 0.1% TFA; (D) formic acid:H(2)O: 2-propanol (1:3:2); and (E) H(2)O:MeOH (2:1). Each protein was found to have a different optimum solvent system for PMF. Generally, better PMF results were obtained with 2,5-DHB. The best PMF results were obtained when all of the mass spectral data for a particular protein digest were convolved. PMID- 14760720 TI - Result-driven strategies for protein identification and quantitation--a way to optimize experimental design and derive reliable results. AB - Uni- or multidimensional microcapillary liquid chromatography (microLC) matrix assisted laser desorption/ionization (MALDI) tandem mass spectrometry (MS/MS) approaches have gained significant attention for quantifying and identifying proteins in complex biological samples. The off-line coupling of microLC with MS quantitation and MS/MS identification methods makes new result-dependent workflows possible. A relational database is used to store the results from multiple high performance liquid chromatography runs, including information about MALDI plate positions, and both peptide and protein quantitations, and identifications. Unlike electrospray methodology, where all the decisions about which peptide to fragment, must be made during peptide fractionations, in the MALDI experiments the samples are effectively "frozen in time". Therefore, additional MS and MS/MS spectra can be acquired, to promote more accurate quantitation or additional identifications until reliable results are derived that meet experimental design criteria. In the case of what can be designated the expression-dependent workflow, quantitation can be detached from identification and only peak pairs with biological relevant expression changes can be selected for further MS/MS analyses. Alternatively, additional MS/MS data can be acquired to confirm tentative peptide mass fingerprint hits in what is designated a search result-dependent workflow. In the MS data-dependent workflow, the goal is to collect as many meaningful spectra as possible by judiciously adjusting the acquisition parameters based on characteristics of the parent masses. This level of sophistication requires the development of innovative algorithms for these three result-dependent workflows that make MS and MS/MS analysis more efficient and also add confidence to experimental results. PMID- 14760721 TI - Common interchange standards for proteomics data: Public availability of tools and schema. AB - The Proteomics Standards Initiative (PSI) aims to define community standards for data representation in proteomics and to facilitate data comparision, exchange and verification. To this end, a Level 1 Molecular Interaction XML data exchange format has been developed which has been accepted for publication and is freely available at the PSI website (http.//psidev.sf.net/). Several major protein interaction databases are already making data available in this format. A draft XML interchange format for mass spectrometry data has been written and is currently undergoing evaluation whilst work is ongoing to develop a proteomics data integration model, MIAPE. PMID- 14760722 TI - Proteomic analysis on structural proteins of Severe Acute Respiratory Syndrome coronavirus. AB - Recently, a new coronavirus was isolated from the lung tissue of autopsy sample and nasal/throat swabs of the patients with Severe Acute Respiratory Syndrome (SARS) and the causative association with SARS was determined. To reveal further the characteristics of the virus and to provide insight about the molecular mechanism of SARS etiology, a proteomic strategy was utilized to identify the structural proteins of SARS coronavirus (SARS-CoV) isolated from Vero E6 cells infected with the BJ-01 strain of the virus. At first, Western blotting with the convalescent sera from SARS patients demonstrated that there were various structural proteins of SARS-CoV in the cultured supernatant of virus infected Vero E6 cells and that nucleocaspid (N) protein had a prominent immunogenicity to the convalescent sera from the patients with SARS, while the immune response of spike (S) protein probably binding with membrane (M) glycoprotein was much weaker. Then, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS PAGE) was used to separate the complex protein constituents, and the strategy of continuous slicing from loading well to the bottom of the gels was utilized to search thoroughly the structural proteins of the virus. The proteins in sliced slots were trypsinized in-gel and identified by mass spectrometry. Three structural proteins named S, N and M proteins of SARS-CoV were uncovered with the sequence coverage of 38.9, 93.1 and 28.1% respectively. Glycosylation modification in S protein was also analyzed and four glycosylation sites were discovered by comparing the mass spectra before and after deglycosylation of the peptides with PNGase F digestion. Matrix-assisted laser desorption/ionization mass spectrometry determination showed that relative molecular weight of intact N protein is 45 929 Da, which is very close to its theoretically calculated molecular weight 45 935 Da based on the amino acid sequence deduced from the genome with the first amino acid methionine at the N-terminus depleted and second, serine, acetylated, indicating that phosphorylation does not happen at all in the predicted phosphorylation sites within infected cells nor in virus particles. Intriguingly, a series of shorter isoforms of N protein was observed by SDS-PAGE and identified by mass spectrometry characterization. For further confirmation of this phenomenon and its related mechanism, recombinant N protein of SARS-CoV was cleaved in vitro by caspase-3 and -6 respectively. The results demonstrated that these shorter isoforms could be the products from cleavage of caspase-3 rather than that of caspase-6. Further, the relationship between the caspase cleavage and the viral infection to the host cell is discussed. PMID- 14760723 TI - Proteomic analysis of the effect of heat stress on hexaploid wheat grain: characterization of heat-responsive proteins from non-prolamins fraction. AB - The effect of heat stress on hexaploid wheat grain proteome was recently analyzed in our previous works. Proteomic tools allowed the characterization of heat responsive proteins of total endosperm, composed mainly of prolamins. The present work completes this study; our aim was to analyze the effect of heat stress on the water-soluble fraction, composed essentially of albumins and globulins. These proteins were separated by two-dimensional electrophoresis (2-DE), visualized by Coomassie Brilliant Blue (CBB) staining and analyzed by Melanie-3 software. Of the 43 heat-changed proteins, 24 were found to be up-regulated whereas 19 spot proteins were down-regulated. All of these proteins were subjected to matrix assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF MS) followed by database searching which allowed the identification of 42 spots. Of these, some were enzymes involved in different metabolic pathways of plants, such as granule-bound starch synthase and glucose-1-phosphate adenyltransferase, involved in the starch synthesis pathway; beta-amylase, involved in carbohydrate metabolism, and the ATP synthase beta-chain that was related to four heat decreased proteins. Moreover, five heat up-regulated proteins showed similarities with small heat shock proteins while three other spots were related to elongation factors or eucaryotic translation initiation factors. Proteins involved in abiotic stresses or in plant defense mechanism were also identified and are discussed. PMID- 14760724 TI - Proteomic analysis of diet-induced hypercholesterolemic mice. AB - We report here a proteomic analysis of differentially expressed liver proteins of both C57BL/6J (B6, atherosclerosis-susceptible strain) and C3H/HeJ mice (C3H, atherosclerosis-resistant strain), which were fed either control or a high-fat enriched atherogenic diet for eight weeks. We observed differential patterns of plasma lipids between the two strains when both were fed atherogenic diets. That is, although low density lipoprotein cholesterol level was highly elevated in both, the levels of total cholesterol and triglyceride in B6 mice were much lower than those in C3H mice when they were fed atherogenic diets. However, the high density lipoprotein cholesterol level was increased in the latter but decreased in the former. Histopathological observation revealed that more prominent lipid droplets were present in B6 mice than in C3H mice, when they were maintained on the atherogenic diets. Proteomic analysis of liver tissues of these two strains showed that a total of 30 proteins were significantly changed in the livers obtained from both strains after being fed the atherogenic diet. Of these, 14 protein spots including carbonic anhydrase III, senescence marker protein 30 and selenium binding protein 2 were differentially changed only in B6 mice, which was also confirmed in part by Western blotting. An additional 16 protein spots including glutathione S-transferase subclass, apolipoprotein E and chaperonin proteins were changed in both strains. We also identified 28 proteins that were differentially expressed in the livers of both B6 and C3H mice, regardless of diet feeding condition. Of these, 4 protein spots in B6 mice and 11 protein spots in C3H mice were up-regulated. Thirteen strain specific protein spots including antioxidant protein 2, apolipoprotein E and apolipoprotein A-I were also detected in different positions in two-dimensional electrophoresis. These results suggest a clear distinction in differential expression of oxidative stress proteins and lipid metabolism related proteins between the two strains in response to atherogenic diet feeding, which might account for their difference in susceptibility to atherogenesis. PMID- 14760725 TI - Copper-associated liver disease: a proteomics study of copper challenge in a sheep model. AB - Sheep display a variant phenotype with respect to their susceptibility to copper and derivative pathology. The North Ronaldsay sheep are acutely sensitive to environmental copper while the Cambridge breed is much more copper-tolerant. A study of protein expression in the liver of the two different breeds of sheep as a result of copper challenge would aid in the understanding of their differing pathophysiologies and contribute to knowledge of copper toxicosis in man. In this initial study, Cambridge breed sheep were challenged with oral copper and liver proteins were analyzed by two-dimensional (2-D) gel electrophoresis. Proteins whose expression pattern was modified by copper exposure were then identified by peptide mass fingerprinting using matrix assisted laser desorption/ionization time of flight (MALDI-TOF) mass spectrometry. In conclusion, the pattern of changes in protein expression were consistent with an early adaptive response to oxidative challenge. This was followed by evidence of an impaired ability of the liver to compensate as copper loading increased, accompanied by oxidative stress induced injury. PMID- 14760726 TI - Proteome analysis reveals elevated serum levels of clusterin in patients with preeclampsia. AB - Preeclampsia is a pregnancy-specific syndrome and a major cause of maternal mortality. The pathophysiology of preeclampsia is unknown, and no proteome analysis of preeclampsia has been reported. We sought to identify proteins associated with preeclampsia using a proteomic technique and performed two dimensional electrophoresis (2-DE) on sera from six patients with preeclampsia and six normal pregnant women, followed by comparison of the SYPRO Ruby-stained 2 DE profiles. A group of overexpressed spots was identified in the limited study set. Overexpressed spots were identified as clusterin by matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS) followed by peptide mass fingerprinting, a protein database search, and Western blot analysis. Additionally, sera of 80 preeclamptic women and 80 normal pregnant women were processed by immunoassay methods to confirm changes in clusterin concentrations quantitatively. Immunoassays showed that clusterin levels in the 80 preeclamptic women were significantly higher than those in the 80 controls (mean +/- SD; 1.62 +/- 0.46 times reference level in preeclamptic women vs. 1.30 +/- 0.46 times reference level in controls, P < 0.001). Proteomic analysis of serum proteins is a promising tool for studying preeclampsia pathophysiology and identifying proteins associated with preeclampsia. PMID- 14760733 TI - Protein Design at the Crossroads of Biotechnology, Chemistry, and Evolution. Proceedings of the 86th International Titisee Conference. Titisee, Germany, 23-27 October 2002. PMID- 14760735 TI - Lipase-specific foldases. AB - Lipases represent the most important class of enzymes used in biotechnology. Many bacteria produce and secrete lipases but the enzymes originating from Pseudomonas and Burkholderia species seem to be particularly useful for a wide variety of different biocatalytic applications. These enzymes are usually encoded in an operon together with a second gene which codes for a lipase-specific foldase, Lif, which is necessary to obtain enzymatically active lipase. A detailed analysis based on amino acid homology has suggested the classification of Lif proteins into four different families and also revealed the presence of a conserved motif, Rx1x2FDY(F/C)L(S/T)A. Recent experimental evidence suggests that Lifs are so-called steric chaperones, which exert their physiological function by lowering energetic barriers during the folding of their cognate lipases, thereby providing essential steric information needed to fold lipases into their enzymatically active conformation. PMID- 14760736 TI - Functional changes in the family of type 3 copper proteins during evolution. PMID- 14760737 TI - Coiled coil domains: stability, specificity, and biological implications. PMID- 14760738 TI - Combinatorial approaches to novel proteins. PMID- 14760739 TI - Consensus design of repeat proteins. AB - Consensus design is a valuable protein-engineering method that is based on statistical information derived from sequence alignments of homologous proteins. Recently, consensus design was adapted to repeat proteins. We discuss the potential of this novel repeat-based approach for the design of consensus repeat proteins and repeat protein libraries and summarize recent results from such experiments. PMID- 14760740 TI - Construction of an artificial receptor protein ("anticalin") based on the human apolipoprotein D. AB - Human apolipoprotein D (ApoD) is a prominent member of the lipocalin family of proteins and transports arachidonic acid and progesterone in various body fluids. Lipocalins share a structurally conserved beta barrel as their central folding unit, which supports a set of four hypervariable loops that form the entrance to the ligand pocket. Based on this structural pattern ApoD was employed as a scaffold for the combinatorial design of artificial receptor proteins termed anticalins. After randomization of 24 amino acids located within the loop region, several ApoD variants were selected against hemoglobin, a biochemically well characterized model target, by using bacterial phagemid display and colony screening. One variant, dubbed HbgA, was further investigated by surface plasmon resonance interaction analysis and found to complex hemoglobin specifically and with a dissociation constant of about 2 microM. While our previous work on the structurally related insect bilin-binding protein was focused on the generation of binding activity towards low-molecular-weight ligands, this study demonstrates for the first time that a lipocalin can also be tailored to recognize a protein target. The fact that even a human member of this protein family has now been successfully recruited for anticalin construction opens the possibility for future application of such engineered lipocalins as target-recognition vehicles in medical therapy. PMID- 14760741 TI - Addressing the challenge of changing the specificity of RNase T1 with rational and evolutionary approaches. AB - Although ribonuclease T1 (RNase T1) is one of the best-characterized proteins with respect to structure and enzymatic action, numerous attempts at altering the specificity of the enzyme to cleave single-stranded RNA at the 3'-side of adenylic instead of guanylic residues by rational approaches have failed so far. Recently we generated and characterized the RNase T1 variant RV with a 7200-fold increase in adenylyl-3',5'-cytidine (ApC)/guanylyl-3',5'-cytidine (GpC) preference, with the guanine-binding loop changed from 41-KYNNYE-46 (wt) to 41 EFRNWN-46. Now we have introduced the asparagine residue at position 46 of the wild-type enzyme as a single-point mutation in variant E46N and in combination with the Y45W exchange also occurring in RV. Both variants show an improved ApC/GpC preference with a 1450-fold increase for E46N and a 2100-fold increase for Y45W/E46N in comparison to wild-type activity. We also addressed the challenge of altering enzyme specificity with an evolutionary approach. We have randomly introduced point mutations into the RNase T1 wild-type gene and into the gene of the variant RV with different mutation rates. Altogether we have screened about 100,000 individual clones for activity on RNase indicator plates; 533 of these clones were active. A significant change in substrate specificity towards an ApC preference could not be observed for any of these active variants; this demonstrated the magnitude of the challenge to alter the specificity of this evolutionary perfected enzyme. PMID- 14760742 TI - Chimeras of the homing endonuclease PI-SceI and the homologous Candida tropicalis intein: a study to explore the possibility of exchanging DNA-binding modules to obtain highly specific endonucleases with altered specificity. AB - Homing endonucleases are extremely specific endodeoxyribonucleases. In vivo, these enzymes confer mobility on their genes by inducing a very specific double strand cut in cognate alleles that lack the cooling sequence for the homing endonuclease; the cellular repair of the double-strand break with the endonuclease-containing allele as a template leads to integration of the endonuclease gene, completing the homing process. As a result of their extreme sequence specificity, homing endonucleases are promising tools for genome engineering. For this purpose, it is desirable to design enzymes with defined new specificities. To analyse which DNA-binding elements are potential candidates for use in the design of enzymes with modified or even new specificity, we produced several chimeric proteins derived from the Saccharomyces cerevisiae VMA1 intein (PI-SceI) and the related Candida tropicalis VMA1 intein. Although the mature Candida intein is devoid of endonucleolytic activity, the exchange of two DNA binding modules of PI-SceI with the homologous elements from the Candida intein results in an active endonuclease. The low sequence homology in these modules indicates that different protein-DNA contacts are responsible for the recognition of related DNA sequences. This flexibility in DNA recognition should, in principle, allow endonucleases to be produced with new specificities useful for genome engineering. PMID- 14760743 TI - Learning from directed evolution: theoretical investigations into cooperative mutations in lipase enantioselectivity. AB - Molecular modeling with classical force-fields has been used to study the reactant complex and the tetrahedral intermediate in lipase-catalyzed ester hydrolysis in 20 enzyme/substrate combinations. The R and S enantiomers of alpha methyldecanoic acid ester served as substrates for the wild-type lipase from Pseudomonas aeruginosa and nine selected mutants. After suitable preparation of initial structures from an available wild-type crystal structure, each system was subjected to 1 ns CHARMM force-field molecular dynamics simulations. The resulting geometric and energetic changes allow interpretation of some experimentally observed effects of mutations, particularly with regard to the "hot spots" at residues 155 and 162. The replacement S155F enhances S enantiopreference through a steric relay involving Leu162. The double mutation S53P + L162G improves S enantioselectivity by creating a new binding pocket for the S enantiomer with an additional stabilizing hydrogen bond to His83. The simulations provide insight into remote and cooperative effects of mutations. PMID- 14760744 TI - Modulating functional loop movements: the role of highly conserved residues in the correlated loop motions. AB - Loop flexibility in enzymes plays a vital role in correctly positioning catalytically important residues. This strong relationship between enzyme flexibility and function provides an opportunity to engineer new substrates and inhibitors. It further allows the design of site-directed mutagenesis experiments to explore enzymatic activity through the control of flexibility of a functional loop. Earlier, we described a novel mechanism in which a small loop triggers the motions of a functional loop in three enzymes (beta-1,4-galactosyltransferase, lipase, and enolase) unrelated in sequence, structure, or function. Here, we further address the question of how the interactions between various flexible loops modulate the movements of the functional loop. We examine beta-1,4 galactosyltransferase as a model system in which a Long loop undergoes a large conformational change (moves in space up to 20 A) upon substrate binding in addition to a small loop (Trp loop) that shows a considerably smaller conformational change. Our molecular-dynamics simulations carried out in implicit and explicit solvent show that, in addition to these two loops, two other neighboring loops are also highly flexible. These loops are in contact with either the Long loop or the Trp loop. Analysis of the covariance of the spatial displacement of the residues reveals that coupled motions occur only in one of these two loops. Sequence analysis indicates that loops correlated in their motions also have highly conserved residues involved in the loop-loop interactions. Further, analysis of crystal structures and simulations in explicit water open the possibility that the Trp loop that triggers the movement of the Long loop in the unbound conformation may also play the same role in the substrate-bound conformation through its contact with the conserved and correlated third loop. Our proposition is supported by the observation that four of the five conserved positions in the third loop are at the interface with the Trp loop. Evolution appears to select residues that drive the functional Long loop to a large conformational change. These observations suggest that altering selected loop-loop interactions might modulate the movements of the functional loop. PMID- 14760745 TI - Adaptation of class-13 alpha-amylases to diverse living conditions. AB - There are currently 35 available nonredundant molecular structures of class-13 alpha-amylases (EC 3.2.1.1), mostly from microbial organisms living under a wide range of environmental conditions. One of the most recent additions has been the first alpha-amylase structure of a hyperthermophilic archaeon [Linden et al., J. Biol. Chem. 2003, 278, 9875-9884]. The structure has been used for comparative analyses with a representative set of three alpha-amylases from thermophilic, mesophilic and psychrophilic sources to identify molecular parameters for environmental adaptation. Our analysis supports generally observed trends such as an increase in structural compactness as well as an increase in salt bridges in order to cope with high-temperature conditions. The two representative thermophilic structures used in this comparative study have independently evolved di-metal centres--not present in the mesophilic and psychrophilic structures--in the vicinity of the active site. These observations may provide impetus for the design of alpha-amylases with improved molecular properties to enhance their utility in biotechnological processes. PMID- 14760746 TI - European Symposium of Bio-Organic Chemistry 2003 (ESBOC): the evolution of catalysis. PMID- 14760749 TI - Induction of tryptase and histamine release from human colon mast cells by IgE dependent or independent mechanisms. AB - AIM: To investigate the tryptase and histamine release ability of human colon mast cells upon IgE dependent or independent activation and the potential mechanisms. METHODS: Enzymatically dispersed cells from human colons were challenged with anti-IgE or calcium ionophore A23187, and the cell supernatants after challenge were collected. Both concentration dependent and time course studies with anti-IgE or calcium ionophore A23187 were performed. Tryptase release was determined with a sandwich ELISA procedure and histamine release was measured using a glass fibre-based fluorometric assay. RESULTS: Both anti-IgE and calcium ionophore were able to induce dose dependent release of histamine from colon mast cells with up to approximately 60% and 25% net histamine release being achieved with 1 microg/mL calcium ionophore and 10 microg/mL anti-IgE, respectively. Dose dependent release of tryptase was also observed with up to approximately 19 ng/mL and 21 ng/mL release of tryptase being achieved with 10 microg/mL anti-IgE and 1 microg/mL calcium ionophore, respectively. Time course study revealed that both tryptase and histamine release from colon mast cells stimulated by anti-IgE initiated within 10 sec and reached their maximum release at 6 min following challenge. Pretreatment of cells with metabolic inhibitors abolished the actions of anti-IgE as well as calcium ionophore. Tryptase and histamine release, particularly that induced by calcium ionophore was inhibited by pretreatment of cells with pertussis toxin. CONCLUSION: Both anti-IgE and calcium ionophore are able to induce significant release of tryptase and histamine from colon mast cells, indicating that this cell type is likely to contribute to the pathogenesis of colitis and other mast cell associated intestinal diseases. PMID- 14760750 TI - Modulation of tryptase secretion from human colon mast cells by histamine. AB - AIM: To investigate the ability of histamine to modulate tryptase release from human colon mast cells and the potential mechanisms. METHODS: Enzymatically dispersed cells from human colons were challenged with histamine, anti-IgE or calcium ionophore A23187 (CI), and the cell supernatants after challenge were collected. Tryptase release was determined with a sandwich ELISA procedure. RESULTS: Histamine at concentrations from 1 ng/mL was able to induce a "bell" shape dose related release of tryptase from colon mast cells. The maximum release of tryptase was approximately 3.5 fold more than spontaneous release. As little as 10 ng/mL histamine showed a similar potency to 10 microg/mL anti-IgE in induction of tryptase release. Histamine induced release of tryptase initiated at 10 s when histamine (100 ng/mL) was added to cells, gradually increased thereafter, and completed at 5 min. Both pertussis toxin or metabolic inhibitors were able to inhibit histamine induced tryptase release. When histamine and anti IgE were added to colon mast cells at the same time, the quantity of tryptase released was similar to that induced by anti-IgE alone. The similar results were observed with CI. However, when various concentrations of histamine were incubated with cells for 20 min before adding anti-IgE or CI, the quantity of tryptase released was similar to that was induced by histamine alone. CONCLUSION: Histamine is a potent activator of human colon mast cells, which represents a novel and pivotal self-amplification mechanism of mast cell degranulation. PMID- 14760751 TI - Activation of human colon mast cells through proteinase activated receptor-2. AB - AIM: To investigate the ability of agonists of PAR-2 to stimulate release of tryptase and histamine from human colon mast cells and the potential mechanisms. METHODS: Enzymatically dispersed cells from human colons were challenged with tc LIGRLO, tc-OLRGIL, SLIGKV, VKGILS, trypsin, anti-IgE or calcium ionophore A23187, and the cell supernatants after challenge were collected. Tryptase release was determined with a sandwich ELISA procedure and histamine release was measured using a glass fibre-based fluorometric assay. RESULTS: Both PAR-2 agonists tc LIGRLO-NH2 and SLIGKV-NH2 were able to induce dose dependent release of tryptase and histamine from colon mast cells. More than 2.5 fold increase in both tryptase and histamine release was provoked by 100 micromol/mL tc-LIGRLO-NH2, in comparison with only 2.0 fold increase being stimulated by SLIGKV-NH2. The reverse peptides tc-OLRGIL-NH2 and VKGILS -NH2 at the concentrations tested had no effect on the release of these two mediators. The maximum tryptase release elicited by tc-LIGRLO-NH2 was similar to that induced by anti-IgE (10 microg/mL) or calcium ionophore (1 microg/mL), though the latter was a more potent stimulus for histamine release. Both histamine and tryptase release in response to tc LIGRLO-NH2 were completed within 3 min. Trypsin at concentrations from 1.0 to 100 microg/mL was capable of provoking a dose dependent release of tryptase as well as histamine with a maximum of 16 ng/mL tryptase and 14 ng/mL histamine release being achieved. An approximately 80% and 70% inhibition of trypsin induced release of tryptase and histamine were observed with SBTI, respectively. Pretreatment of cells with metabolic inhibitors or pertussis toxin abolished the actions of tc-LIGRLO-NH2, SLIGKV-NH2 and trypsin. CONCLUSION: The agonists of PAR 2 and trypsin are potent secretagogues of human colon mast cells, which are likely to contribute to the development of inflammatory disorders in human gut. PMID- 14760748 TI - Key role of mast cells and their major secretory products in inflammatory bowel disease. AB - Historically, mast cells were known as a key cell type involved in type I hypersensitivity. Until last two decades, this cell type was recognized to be widely involved in a number of non-allergic diseases including inflammatory bowel disease (IBD). Markedly increased numbers of mast cells were observed in the mucosa of the ileum and colon of patients with IBD, which was accompanied by great changes of the content in mast cells such as dramatically increased expression of TNFalpha, IL-16 and substance P. The evidence of mast cell degranulation was found in the wall of intestine from patients with IBD with immunohistochemistry technique. The highly elevated histamine and tryptase levels were detected in mucosa of patients with IBD, strongly suggesting that mast cell degranulation is involved in the pathogenesis of IBD. However, little is known of the actions of histamine, tryptase, chymase and carboxypeptidase in IBD. Over the last decade, heparin has been used to treat IBD in clinical practice. The low molecular weight heparin (LMWH) was effective as adjuvant therapy, and the patients showed good clinical and laboratory response with no serious adverse effects. The roles of PGD2, LTC4, PAF and mast cell cytokines in IBD were also discussed. Recently, a series of experiments with dispersed colon mast cells suggested there should be at least two pathways in man for mast cells to amplify their own activation-degranulation signals in an autocrine or paracrine manner. The hypothesis is that mast cell secretogogues induce mast cell degranulation, release histamine, then stimulate the adjacent mast cells or positively feedback to further stimulate its host mast cells through H1 receptor. Whereas released tryptase acts similarly to histamine, but activates mast cells through its receptor PAR-2. The connections between current anti-IBD therapies or potential therapies for IBD with mast cells were discussed, implicating further that mast cell is a key cell type that is involved in the pathogenesis of IBD. In conclusion, while pathogenesis of IBD remains unclear, the key role of mast cells in this group of diseases demonstrated in the current review implicates strongly that IBD is a mast cell associated disease. Therefore, close attentions should be paid to the role of mast cells in IBD. PMID- 14760752 TI - Inhibition of tryptase release from human colon mast cells by protease inhibitors. AB - AIM: To investigate the ability of protease inhibitors to modulate tryptase release from human colon mast cells. METHODS: Enzymatically dispersed cells from human colon were challenged with anti-IgE or calcium ionophore A23187 in the absence or presence of tryptase and chymase inhibitors, and tryptase release was determined. RESULTS: IgE dependent tryptase release from colon mast cells was inhibited by up to approximately 37%, 40% and 36.6% by chymase inhibitors Z-Ile Glu-Pro-Phe-CO2Me (ZIGPFM), N-tosyl-L-phenylalanyl-chloromethyl ketone (TPCK), and alpha1-antitrypsin, respectively. Similarly, the inhibitors of tryptase leupeptin, N-tosyl-L-lysine chloromethyl ketone (TLCK) and lactoferrin were also able to inhibit anti-IgE induced tryptase release by a maximum of 39.4%, 47.6% and 36.6%, respectively. The inhibitory actions of chymase inhibitors, but not tryptase inhibitors on colon mast cells were enhanced by preincubation of them with cells for 20 min before challenged with anti-IgE. At a concentration of 10 microg/mL, protamine was able to inhibit anti-IgE and calcium ionophore induced tryptase release. However, at 100 microg/mL, protamine elevated tryptase levels in supernatants. A specific inhibitor of aminopeptidase amastatin had no effect on anti-IgE induced tryptase release. The significant inhibition of calcium ionophore induced tryptase release was also observed with the inhibitors of tryptase and chymase examined. The inhibitors tested by themselves did not stimulate tryptase release from colon mast cells. CONCLUSION: It was demonstrated for the first time that both tryptase and chymase inhibitors could inhibit IgE dependent and calcium ionophore induced tryptase release from dispersed colon mast cells in a concentration dependent of manner, which suggest that they are likely to be developed as a novel class of anti-inflammatory drugs to treat chronic of colitis in man. PMID- 14760753 TI - Modulation of histamine release from human colon mast cells by protease inhibitors. AB - AIM: To investigate the ability of protease inhibitors to modulate histamine release from human colon mast cells. METHODS: Enzymatically dispersed cells from human colon were challenged with anti-IgE or calcium ionophore A23187 in the absence or presence of tryptase and chymase inhibitors, and histamine release was determined. RESULTS: IgE dependent histamine release from colon mast cells was inhibited by up to approximately 37%, 26% and 36.8% by chymase inhibitors Z-Ile Glu-Pro-Phe-CO2Me (ZIGPFM), N-Tosyl-L-phenylalanyl-chloromethyl ketone (TPCK), and alpha1-antitrypsin, respectively. Similarly, inhibitors of tryptase leupeptin, N-tosyl-L-lysine chloromethyl ketone (TLCK), lactoferrin and protamine were also able to inhibit anti-IgE induced histamine release by a maximum of some 48%, 37%, 40% and 34%, respectively. Preincubation of these inhibitors with cells for 20 min before challenged with anti-IgE had small effect on the inhibitory actions of these inhibitors on colon mast cells. A specific inhibitor of aminopeptidase amastatin had no effect on anti-IgE induced histamine release. The significant inhibition of calcium ionophore induced histamine release was also observed with the inhibitors of tryptase and chymase examined. Apart from leupeptin and protamine, the inhibitors tested by themselves did not stimulate colon mast cells. CONCLUSION: It was demonstrated that both tryptase and chymase inhibitors could inhibit IgE dependent and calcium ionophore induced histamine release from dispersed colon mast cells in a concentration dependent of manner, which suggest that they are likely to be developed as a novel class of anti inflammatory drugs to treat chronic of colitis in man. PMID- 14760754 TI - Cloning and expression of human colon mast cell carboxypeptidase. AB - AIM: To clone and express the human colon mast cell carboxypeptidase (MC-CP) gene. METHODS: Total RNA was extracted from colon tissue, and the cDNA encoding human colon mast cell carboxypeptidase was amplified by reverse-transcription PCR (RT-PCR). The product cDNA was subcloned into the prokaryotic expression vector pMAL-c2x and eukaryotic expression vector pPIC9K to construct prokaryotic expression vector pMAL/human MC-CP (hMC-CP) and eukaryotic pPIC9K/hMC-CP. The recombinant fusion protein expressed in E. coli was induced with IPTG and purified by amylose affinity chromatography. After digestion with factor Xa, recombinant hMC-CP was purified by heparin agarose chromatography. The recombinant hMC-CP expressed in Pichia pastoris (P. pastoris) was induced with methanol and analyzed by SDS-PAGE, Western blot, N-terminal amino acid sequencing and enzyme assay. RESULTS: The cDNA encoding the human colon mast cell carboxypeptidase was cloned, which had five nucleotide variations compared with skin MC-CP cDNA. The recombinant hMC-CP protein expressed in E. coli was purified with amylose affinity chromatography and heparin agarose chromatography. SDS-PAGE and Western blot analysis showed that the recombinant protein expressed by E. coli had a molecular weight of 36 kDa and reacted to the anti-native hMC-CP monoclonal antibody (CA5). The N-terminal amino acid sequence confirmed further the product was hMC-CP. E. coli generated hMC-CP showed a very low level of enzymatic activity, but P. pastoris produced hMC-CP had a relatively high enzymatic activity towards a synthetic substrate hippuryl-L-phenylalanine. CONCLUSION: The cDNA encoding human colon mast cell carboxypeptidase can be successfully cloned and expressed in E. coli and P. pastoris, which will contribute greatly to the functional study on hMC-CP. PMID- 14760755 TI - Apoptosis of human gastric adenocarcinoma cells induced by beta-ionone. AB - AIM: To investigate the effect of beta-ionone on the growth and apoptosis of gastric adenocarcinoma cell line SGC-7901. METHODS: Using MTT, fluorescence dye (Hoechst-33258), transmission electron microscopy and the TUNEL assay, we examined growth and apoptosis of SGC-7901 cells treated with beta-ionone at various concentrations (i.e. 25, 50, 100 and 200 micromol/L) for 24 h, 48 h. RESULTS: The growth of SGC-7901 cells was inhibited by beta-ionone. Seven days after treatment with beta-ionone at four concentrations, the inhibition rates were 12.04%, 30.59%, 78.25% and 94.15%, respectively. The IC(50) value of beta ionone for SGC-7901 cells was estimated to be 89 micromol/L. The apoptotic morphology was demonstrated in SGC-7901 cells treated with beta-ionone by Hoechst 33258 staining and electron microscopy. Apoptosis was also shown in beta-ionone treated SGC-7901 cells by the TUNEL assay. CONCLUSION: beta-ionone can inhibit cell proliferation and induce apoptosis of SGC-7901 cells. However, the mechanism needs to be further investigated. PMID- 14760756 TI - Expression and significance of VEGF-C and FLT-4 in gastric cancer. AB - AIM: To investigate the expression of pathological factors of VEGF-C and its receptor FLT-4 in primary gastric cancer and adjacent normal tissues. METHODS: The expression of VEGF-C and FLT-4 was studied in 80 primary gastric cancers and adjacent normal tissues from the same patients by semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) and immumohistochemistry. RESULTS: Both primary gastric cancer and adjacent normal tissue could express VEGF-C and FLT-4, and FLT-4 expression was also detected in endothelial cells of stromal blood vessels and lymphatic vessels. There was a significant difference in expression of VEGF-C between primary tumor and adjacent normal tissue samples (P=0.01), and a statistical correlation between VEGF-C and FLT-4 expression in tumors (P=0.00886). With regard to VEGF-C expression, there was a significant difference between moderate-poor differential type and high differential type (P=0.032), and a significant difference between positive and negative lymph node metastases (P=0.024). However, there was no significant difference between positive and negative serosal invasions (P=0.219). CONCLUSION: VEGF-C and its receptor FLT-4 play a role in the development of gastric cancer, and the tumors with expression of VEGF-C and FLT-4 are more likely to have lymph node metastasis. PMID- 14760757 TI - Activating mechanism of transcriptor NF-kappaB regulated by hepatitis B virus X protein in hepatocellular carcinoma. AB - AIM: To investigate the mechanism and significance of NF-kappaB activation regulated by hepatitis B virus X protein (HBx) in hepatitis B virus (HBV) associated hepatocellular carcinoma (HCC). METHODS: The expression levels of HBx, p65, IkappaB-alpha and ubiquitin were detected by immunohistochemistry in HCC tissue microarrays (TMA) respectively, and IkappaB-alpha was detected by Western blot in HCC and corresponding liver tissues. RESULTS: The percentage of informative TMA samples was 98.8% in 186 cases with a total of 367 samples. Compared with corresponding liver tissues (60.0%), the HBx expression was obviously decreased in HBV-associated HCC (47.9%, u=2.24, P<0.05). On the contrary, the expressions of p65 (20.6% vs 45.3%, u=4.85, P<0.01) and ubiquitin (8.9% vs 59.0%, u=9.68, P<0.01) were notably elevated in HCC. In addition, IkappaB-alpha had a tendency to go up. Importantly, positive relativity was observed between HBx and p65 (chi2=10.26, P<0.01), p65 and IkappaB-alpha (chi (2)=16.86, P<0.01), IkappaB-alpha and ubiquitin (chi2=8.90, P<0.01) in HCC, respectively. CONCLUSION: Both active and non-active forms of NF-kappaB are increased in HBV-associated HCC. Variant HBx is the major cause of the enhancement of NF-kappaB activity. The activation always proceeds in nucleus and the proteasome complexes play an important role in the activation. PMID- 14760758 TI - Purification of heat shock protein 70-associated tumor peptides and their antitumor immunity to hepatoma in mice. AB - AIM: To purify the heat shock protein (HSP) 70-associated tumor peptides and to observe its non-MHC-I molecule restrictive antitumor effect. METHODS: By ConA sepharose affinity chromatography, ADP-agarose affinity chromatography, and DEAE anion exchange chromatography, we were able to purify HSP70-associated peptides from mouse hepatoma (HCaF) cells treated in heat shock at 42 degrees. Specific active immunization and adoptive cellular immunization assay were adopted to observe the immunoprotective effect elicited by HSP70-associated peptide complexes isolated from HcaF. RESULTS: The finally purified HSP-associated peptides had a very high purity and specificity found by SDS-PAGE and Western blot. Mice immunized with HSP70-associated peptide complexes purified from HCaF cells were protected from HCaF living cell challenge. This effect was dose dependent. Adoptive immunization of immune spleen cells of mice immunized with HSP70-associated peptide complexes could elicit immunity against HCaF challenge, and the tumor-free mice could resist repeated challenges. This effect could be continuously enhanced by repeated challenge with HCaF living cells. The tumor free mice could tolerate the challenge for as high as 1 x 10(7) HCaF cells. The mice immunized once with spleen cells pulsed with HSP70-associated peptide complexes in vitro could also result in a certain adoptive immunity against HCaF. CONCLUSION: High purity and specificity of HSP70-associated peptides could be achieved from tumor cells by the low-pressure affinity chromatography method used in this study. HSP70-associated peptide complexes derived from the HCaF can elicit non-MHC-I molecule restrictive immunoprotective effect against HCaF. This effect can be transferred by adoptive immunization to mice and enhanced by repeated challenge with HCaF live cells. PMID- 14760759 TI - Antisense oligonucleotide targeting at the initiator of hTERT arrests growth of hepatoma cells. AB - AIM: To evaluate the inhibitory effect of antisense phosphorothioate oligonucleotide (asON) complementary to the initiator of human telomerase catalytic subunit (hTERT) on the growth of hepatoma cells. METHODS: The as-hTERT was synthesized by using a DNA synthesizer. HepG2.2.15 cells were treated with as hTERT at the concentration of 10 micromol/L. After 72 h, these cells were obtained for detecting growth inhibition, telomerase activity using the methods of MTT, TRAP-PCR-ELISA, respectively. BALB/c(nu/nu) mice were injected HepG2.2.15 cells and a human-nude mice model was obtained. There were three groups for anti tumor activity study. Once tumors were established, these animals in the first group were administered as-hTERT and saline. Apoptosis of tumor cells was detected by FCM. In the 2nd group, the animals were injected HepG2.2.15 cells together with as-hTERT. In the third group, the animals were given as-hTERT 24 hours postinjection of HepG2.2.15 cells. The anti-HBV effects were assayed with ELISA in vitro and in vivo. RESULTS: Growth inhibition was observed in cells treated with as-hTERT in vitro. A significant different in the value of A570-A630 was found between cells treated with as-hTERT and control (P<0.01) by MTT method. The telomerase activity of tumor cells treated with as-hTERT was reduced, the value of A450 nm was 0.42 compared to control (1.49) with TRAP-PCR-ELISA. The peak of apoptosis in tumor cells given as-hTERT was 21.12%, but not seen in saline-treated control. A prolonged period of carcinogenesis was observed in the second and third group animals. There was inhibitory effect on the expression of HBsAg and HBeAg in vivo and in vitro. CONCLUSION: As-hTERT has an anti-tumor activity, which may be useful for gene therapy of tumors. PMID- 14760760 TI - Nuclear and mitochondrial DNA microsatellite instability in hepatocellular carcinoma in Chinese. AB - AIM: To study the nuclear microsatellite instability (nMSI) at BAT26 and mitochondrial microsatellite instability (mtMSI) in the occurrence and development of hepatocellular carcinoma and the relationship between nMSI and mtMSI. METHODS: nMSI was observed with PCR and mtMSI with PCR-SSCP in 52 cases of hepatocellular carcinoma. RESULTS: mtMSI was detected in 11 out of the 52 cases of hepatocellular carcinoma (21.2%). Among the 11 cases of hepatocellular carcinoma with mtMSI, 7 occurred in one locus and 4 in 2 loci. The frequency of mtMSI in the 52 cases of hepatocellular carcinoma showed no correlation to sex, age, infection of hepatitis B, liver cirrhosis as well as positive AFP of the patients (P>0.05). In addition, nMSI was detected in 3 out of 52 cases of hepatocellular carcinoma (5.8%) and there was no correlation of the incidence of mtMSI to that of nMSI (P>0.05). CONCLUSION: mtMSI may be involved in the coccurrence and development of hepatocellular carcinoma and it is independent of nMSI. PMID- 14760761 TI - Hemizygous deletion and hypermethylation of RUNX3 gene in hepatocellular carcinoma. AB - AIM: To analyze the genetic and epigenetic alterations of RUNX3 gene, a potential putative tumor suppressor gene, in hepatocellular carcinoma (HCC). METHODS: PCR based loss of heterozygosity (LOH) detection, analysis of mutation with PCR single strand conformational polymorphism (SSCP) and sequencing, and methylation study with methylation specific PCR (MSP) were performed on RUNX3 gene in a series of 62 HCCs along with their matched normal tissues. RESULTS: Mutation of RUNX3 gene was not found, but one single nucleotide polymorphism with T to A transversion at the second nucleotide of the 18th codon was found. Nine of 26 informative cases (34.6%) showed allelic loss on the polymorphic site and 30 cases (48.4%) revealed hypermethylation of RUNX3 gene in promoter CpG islands. Furthermore, of the 9 cases with LOH, 8 (88.9%) also had hypermethylation. CONCLUSION: Our findings indicate that inactivation of RUNX3 gene through allelic loss and promoter hypermethylation might be one of the major mechanisms in hepatocellular carcinogenesis. PMID- 14760762 TI - Values of high-resolution computed tomography and pulmonary function tests in managements of patients with chronic hepatitis C virus infection. AB - AIM: To investigate pulmonary involvement via pulmonary function tests (PFT) and high-resolution computed tomography (HRCT) in patients with chronic hepatitis C virus (HCV) infection. METHODS: Thirty-four patients with chronic HCV infection without diagnosis of any pulmonary diseases and 10 healthy cases were enrolled in the study. PFT and HRCT were performed in all cases. RESULTS: A decrease lower than 80% of the predicted value was detected in vital capacity in 9/34 patients, in forced expiratory volume in one second in 8/34 patients, and in forced expiratory flow 25-75 in 15/34 patients, respectively. Carbon monoxide diffusing capacity (DLCO) was decreased in 26/34 patients. Findings of interstitial pulmonary involvement were detected in the HRCT of 16/34 patients. Significant difference was found between controls and patients with HCV infection in findings of HRCT (chi2=4.7, P=0.003). Knodell histological activity index (KHAI) of 28/34 patients in whom liver biopsy was applied was 9.0+/-4.7. HRCT findings, PFT values and DLCO were not affected by KHAI in patients with HCV infection. In these patients, all the parameters were related with age. CONCLUSION: We suggest that chronic hepatitis C virus infection may cause pulmonary interstitial involvement without evident respiratory symptoms. PMID- 14760763 TI - Expression of platelet-derived growth factor-BB in liver tissues of patients with chronic hepatitis B. AB - AIM: To study the relationship between expression of platelet-derived growth factor-BB (PDGF-BB) and fibrogenesis in chronic hepatitis B. METHODS: Hepatic tissues from 43 patients with chronic hepatitis B were embedded in paraffin. The sections were stained with HE and picric acid-sirius red to determine inflammatory activity and fibrosis stages. PDGF-BB expression was detected by immunohistochemistry and assessed semiquantatively. Levels of serum hyaluronic acid (HA), pro-collagen III (PCIII), collagen IV (IV-C) and laminin (LN) were examined by radioimmunoassay (RIA). RESULTS: The expression level of PDGF-BB was found to be positively correlated with inflammatory activity, fibrosis stage and grade of histological findings (tau=0.58, 0.55, 0.55, P<0.01). The positive correlation was also observed between tissue level of PDGF-BB expression and contents of HA, PCIII, IV-C and LN in the circulation (r=0.52, 0.32, 0.40, 0.33, P<0.05). CONCLUSION: PDGF-BB may play some role in the development and progression of liver fibrosis. PMID- 14760764 TI - Viral replication modulated by synthetic peptide derived from hepatitis B virus X protein. AB - AIM: A strategy for viral vaccine design is the use of conserved peptides to overcome the problem of sequence diversity. At present it is still unclear whether conserved peptide is safe as a candidate vaccine. We reported it here for the first time not only to highlight the biohazard issue and safety importance for viral peptide vaccine, but also to explore the effect of a fully conserved peptide on HBV replication within the carboxyl terminus of HBx. METHODS: We synthesized the fully conserved peptide (CP) with nine residues, FVLGGCRHK. HBV producing 2.2.15 cells were treated with or without 3.5 microM CP for 36 hours. Quantitative detection of viral DNA was performed by real-time PCR. HBV antigens were determined by enzyme-linked immunoadsorbent assay (ELISA). Quantitative analyses of p53 and Bax proteins were based on immunofluorescence. Flow cytometry was performed to detect cell cycle and apoptosis. RESULTS: Both extracellular and intracellular copies of HBV DNA per ml were significantly increased after incubation with 3.5 microM of CP. HBsAg and HBeAg in the cultured medium of CP treatment cells were as abundant as untreated control cells. CP influenced negatively the extracellular viral gene products, and 3.5 microM CP could significantly inhibit intracellular HBsAg expression. In response to CP, intracellular HBeAg displayed an opposite pattern to that of HBsAg, and 3.5 microM CP could efficiently increase the level of intracellular HBeAg. Flow cytometric analyses exhibited no significant changes on cell cycle, apoptosis, p53 and Bax proteins in 2.2.15 cells with or without CP. CONCLUSION: Together with the results generated from the synthetic peptide, we address that the conserved region, a domain of HBx, may be responsible for modulating HBV replication. As conserved peptides from infectious microbes are used as immunogens to elicit immune responses, their latent biological hazard for human beings should be evaluated. PMID- 14760765 TI - Antiangiogenic effect of somatostatin receptor subtype 2 on pancreatic cancer cell line: Inhibition of vascular endothelial growth factor and matrix metalloproteinase-2 expression in vitro. AB - AIM: To investigate the anti-angiogenic effect of somatostatin receptor subtype 2 (SSTR2) gene transfer into pancreatic cancer cell line PC-3, and the mechanisms involved in this effect. METHODS: The full length human SSTR2 cDNA was introduced into pancreatic cancer cell line PC-3 by lipofectamine-mediated transfection. Positive clones were screened by G418 and stable expression of SSTR2 was detected by immunohistochemistry SABC methods and RT-PCR. Enzyme-linked immunosorbent assay (ELISA) was used to detect vascular endothelial growth factor (VEGF) levels in the cell culture supernatants of SSTR2-expressing cells, vector control and mock control cells. Furthermore, the expressions of VEGF and matrix metalloproteinase-2 (MMP-2) were detected by immunohistochemistry SABC methods and RT-PCR in these cells. RESULTS: VEGF levels in the cell culture supernatants were significantly reduced in the SSTR2-expressing cells (first week, 172.63+/ 21.2 ng/L and after two months, 198.85+/-26.44 ng/L) compared with the vector control (first week, 790.39+/-86.52 ng/L and after two months, 795.69+/-72.35 ng/L) and mock control (first week, 786.42+/-90.62 ng/L and after two months, 805.32+/-84.36 ng/L) (P<0.05). The immunohistochemical assay showed a significant reduction of the integral optical density of VEGF and MMP-2 in the SSTR2 expressing cells (42.25+/-8.6 and 70.5+/-6.25, respectively) compared with the vector control (85.75+/-12.9 and 110.52+/-13.5, respectively) and mock control (82.6+/-9.28 and 113.56+/-9.62, respectively) (P<0.05). Conversely, the average gray value of VEGF and MMP-2 was significantly increased in the SSTR2-expressing cells (121.56+/-8.43 and 134.46+/-19.95, respectively) compared with the vector control (55.72+/-5.6 and 62.26+/-12.68, respectively) and mock control cells (58.48+/-6.2 and 65.49+/-9.16, respectively) (P<0.05). Moreover, the expressions of VEGF mRNA and MMP-2 mRNA were significantly reduced in the SSTR2-expressing cells (0.1384+/-0.017 and 0.2343+/-0.070, respectively) compared with the vector control (1.024+/-0.117 and 0.806+/-0.119, respectively) and mock control (1.085+/ 0.105 and 0.714+/-0.079, respectively) (P<0.05). CONCLUSION: The expression of reintroduced human SSTR2 gene exerts its antiangiogenic effects by down regulating the expressions of the factors involved in tumor angiogenesis and metastasis, suggesting SSTR2 gene transfer as a new strategy of gene therapy for pancreatic cancer. PMID- 14760766 TI - Herpes simplex virus thymidine kinase and ganciclovir suicide gene therapy for human pancreatic cancer. AB - AIM: To investigate the in vitro effects of suicide gene therapy system of herpes simplex virus thymidine kinase gene (HSV-TK) in combination with the treatment of nucleotide analog-ganciclovir (GCV) on human pancreatic cancer, and to provide a novel clinical therapeutic method for human pancreatic cancer. METHODS: We used a replication defective recombinant retrovirus vector GINaTK (bearing HSV-TK gene) to make packaging cell PA317 produce progeny virions. We then transferred the HSV TK gene to target cells SW1990 using these progeny virions, and treated these gene-modified tumor cells with GCV to study the sensitivity of the cells to GCV and their bystander effects by routine MTT-method. RESULTS: Packaging cell PA317/TK was successfully constructed, and we acquired SW1990/TK through virus progeny infection. These gene-modified pancreatic cancer cells were sensitive to the treatment of GCV compared with unmodified tumor cells (t=4.15, n=10, P<0.0025). We also observed a remarkable bystander effect by mixing two kinds of cells at different ratio. CONCLUSION: Our data demonstrate that HSV-TK/GCV suicide gene therapy system is effective for treating experimental human pancreatic cancer, which is largely resistant to the common therapies, so the suicide gene therapy system may be a potential treatment approach for pancreatic cancer. PMID- 14760767 TI - Striking elevation in incidence and prevalence of inflammatory bowel disease in a province of western Hungary between 1977-2001. AB - AIM: An investigation into inflammatory bowel disease and colorectal cancer in Veszprem Province was conducted from 1977 to 2001. METHODS: Both hospital and outpatient records were collected and reviewed comprehensively. The majority of patients were followed up regularly. RESULTS: The population of the province was decreased from 386,000 to 376,000 during the period. Five hundred sixty new cases of ulcerative colitis (UC), 212 of Crohn's disease (CD), and 40 of indeterminate colitis (IC) were diagnosed. The incidence rates increased from 1.66 to 11.01 cases per 100,000 persons for UC, from 0.41 to 4.68 for CD and from 0.26 to 0.74 for IC. The prevalence rate at the end of 2001 was 142.6 for UC and 52.9 cases per 100,000 persons for CD. The peak onset age in UC patients was between 30 and 40 years, in CD between 20 and 30 years. A family history of IBD was present in 3.4 % in UC and 9.9 % in CD patients. Smoking increased the risk for CD (OR=1.98) while it decreased the risk for UC (OR=0.25). Twelve colorectal carcinomas were observed in this cohort, the cumulative colorectal cancer risk after 10 years in UC was 2%, after 20 years 8.8%, after 30 years 13.3%. CONCLUSION: The incidence and prevalence rates of IBD have increased steadily in Veszprem Province, now equivalent to that in Western European countries. Rapid increase in incidence rates supports a probable role for environmental factors. The rate of colorectal cancers in IBD is similar to that observed in Western countries. PMID- 14760768 TI - Comparison of different intervention procedures in benign stricture of gastrointestinal tract. AB - AIM: To determine the most effective intervention procedure by evaluation of mid and long-term therapeutic efficacy in patients of stricture of the gastrointestinal tract (GIT). METHODS: Different intervention procedures were used to treat benign stricture of GIT in 180 patients including pneumatic dilation (group A, n=80), permanent (group B, n=25) and temporary (group C, n=75) placement of expandable metallic stents. RESULTS: The diameters of the strictured GIT were significantly greater after the treatment of all procedures employed (P<0.01). For the 80 patients in group A, 160 dilations were performed (mean, 2.0 times per patient). Complications in group A included chest pain (n=20), reflux (n=16), and bleeding (n=6). Dysphagia relapse occurred in 24 (30%) and 48 (60%) patients respectively during 6-and-12 month follow-up periods in group A. In group B, 25 uncovered or partially covered or antireflux covered expandable metallic stents were placed permanently, complications included chest pain (n=10), reflux (n=15), bleeding (n=3), and stent migration (n=4), and dysphagia relapse occurred in 5 (20%) and 3 patients (25%) during the 6-and-12 month follow up periods, respectively. In group C, the partially covered expandable metallic stents were temporarily placed in 75 patients and removed after 3 to 7 days via gastroscope, complications including chest pain (n=30), reflux (n=9), and bleeding (n=12), and dysphagia relapse occurred in 9 (12%) and 8 patients (16%) during the 6-and-12 month follow-up periods, respectively. The placement and withdrawal of stents were all successfully performed. The follow-up of all patients lasted for 6 to 96 months (mean 45.3+/-18.6 months). CONCLUSION: The effective procedures for benign GIT stricture are pneumatic dilation and temporary placement of partially-covered expandable metallic stents. Temporary placement of partially-covered expandable metallic stents is one of the best methods for benign GIT strictures in mid and long-term therapeutic efficacy. PMID- 14760769 TI - Survivin expression induced by doxorubicin in cholangiocarcinoma. AB - AIM: To study the role of survivin expression induced by chemotherapy agent (doxorubicin) in the development and anti-chemotherapy of cholangiocarcinoma. METHODS: Expression of survivin was detected by SP immunohistochemical technique in 33 cases of cholangiocarcinoma, 28 cases of adjacent noncancerous bile duct, and 5 cases of benign bile duct lesions. Low concentration of doxorubicin (0.05 mg/l) was added in cultured cholangiocarcinoma cell line (QBC939). The expression of survivin was detected by RT-PCR and Western blot at 24 h and 48 h after adding doxorubicin. RESULTS: Survivin was expressed in 24 of 33 cholangiocarcinoma cases (72.7%). In contrast, no expression of survivin in adjacent noncancerous and benign bile duct lesions was observed (P<0.01). No correlation was found between survivin expression and clinical features. Doxorubicin could markedly (P<0.001) up-regulate survivin mRNA and protein expression of QBC939 cells. CONCLUSION: Overexpression of survivin in cholangiocarcinomas may play an important role in the development of cholangiocarcinoma, its relationship with prognosis of cholangiocarcinoma deserves further investigation. Higher expression of survivin is induced by doxorubicin in QBC939. Survivin expression may resist apoptosis induced by chemotherapy agents. PMID- 14760770 TI - Effects of gastric pacing on gastric emptying and plasma motilin. AB - AIM: To investigate the effects of gastric pacing on gastric emptying and plasma motilin level in a canine model of gastric motility disorders and the correlation between gastric emptying and plasma motilin level. METHODS: Ten healthy Mongrel dogs were divided into: experimental group of six dogs and control group of four dogs. A model of gastric motility disorders was established in the experimental group undergone truncal vagotomy combined with injection of glucagon. Gastric half-emptying time (GEt(1/2)) was monitored with single photon emission computerized tomography (SPECT), and the half-solid test meal was labeled with an isotope-(99m)Tc sulfur colloid. Plasma motilin concentration was measured with radioimmunoassay (RIA) kit. Surface gastric pacing at 1.1-1.2 times the intrinsic slow-wave frequency and a superimposed series of high frequency pulses (10-30 Hz) was performed for 45 min daily for a month in conscious dogs. RESULTS: After surgery, GEt(1/2) in dogs undergone truncal vagotomy was increased significantly from 56.35+/-2.99 min to 79.42+/-1.91 min (P<0.001), but surface gastric pacing markedly accelerated gastric emptying and significantly decreased GEt(1/2) to 64.94+/-1.75 min (P<0.001) in animals undergone vagotomy. There was a significant increase of plasma level of motilin at the phase of IMCIII (interdigestive myoelectrical complex, IMCIII) in the dogs undergone bilateral truncal vagotomy (baseline vs vagotomy, 184.29+/-9.81 pg/ml vs 242.09+/-17.22 pg/ml; P<0.01). But plasma motilin concentration (212.55+/-11.20 pg/ml; P<0.02) was decreased significantly after a long-term treatment with gastric pacing. Before gastric pacing, GEt(1/2) and plasma motilin concentration of the dogs undergone vagotomy showed a positive correlation (r=0.867, P<0.01), but after a long-term gastric pacing, GEt(1/2) and motilin level showed a negative correlation (r=-0.733, P<0.04). CONCLUSION: Surface gastric pacing with optimal pacing parameters can improve gastric emptying parameters and significantly accelerate gastric emptying and can resume or alter motor function in a canine model of motility disorders. Gastric emptying is correlated well with plasma motilin level before and after pacing, which suggests that motilin can modulate the mechanism of gastric pacing by altering gastric motility. PMID- 14760771 TI - Surgical salvage therapy of anal cancer. AB - AIM: To evaluate the results of salvage resection in the management of persistent or locally recurrent anal canal cancer. METHODS: Details of all patients with anal canal cancer treated from 1978 to 1994 at Cancer Hospital of Chinese Academy of Medical Sciences (CAMS) were reviewed retrospectively. Sixteen patients who presented with persistent or locally recurrent anal canal cancer received salvage surgery. Before surgery all of the patients had received radiotherapy alone as their primary treatments. RESULTS: Of the 16 patients, 14 received salvage abdominoperineal resection (APR) and two had transanal local excision. There were no deaths attributable to operation. Delayed healing of the perineal wound occurred in eight patients. Complications unrelated to the perineal wound were found in five patients. The median follow-up time was 120 (range 5-245) months after salvage surgery. Nine patients died of disease progression, with a median survival time of 16 (range 5-27) months. Six patients had a long-term survival. CONCLUSION: Salvage resection after radiotherapy can yield a long-time survival in selected patients with anal canal cancer. However it offers little hope to patients with T4 and/or N(2-3) tumors. PMID- 14760772 TI - Utility of serum CA19-9 in diagnosis of cholangiocarcinoma: in comparison with CEA. AB - AIM: The diagnosis of cholangiocarcinoma is often difficult, making management approaches problematic. A reliable serum marker for cholangiocarcinoma would be a useful diagnostic test. The aims of our study were to evaluate the usefulness of a serum CA19-9 determination in the diagnosis of cholangiocarcinoma. METHODS: We prospectively measured serum CA19-9 and CEA concentrations in patients with cholangiocarcinoma (n=35), benign biliary diseases (n=92), and healthy individuals (n=15). Serum CA19-9 and CEA concentrations were measured by an immunoradiometric assay without knowledge of the clinical diagnosis. RESULTS: The sensitivity of a CA19-9 value >37 KU/L(-1) and a CEA value >22 microg/L(-1) in diagnosing cholangiocarcinoma were 77.14% and 68.57%, respectively. When compared with the benign biliary diseases group, the true negative rates of serum CA19-9 and CEA were 84.78% and 81.52%, respectively. The false positive rates of serum CA19-9 and CEA were 15.22% and 18.48%, whereas the accuracy of serum CA19-9 and CEA were 82.68% and 77.95%, respectively. Serum CA19-9 and CEA concentrations were significantly elevated (P<0.001 and P<0.05) in patients with cholangiocarcinoma (290.31+/-5.34 KU/L(-1) and 36.46+/-18.03 microg/L(-1)) compared with patients with benign biliary diseases (13.38+/-2.59 KU/L(-1) and 13.84+/-3.85 microg/L(-1)) and healthy individuals (12.78+/-3.69 KU/L(-1) and 11.48+/-3.37 microg/L(-1)). In 15 patients undergoing curative resection of cholangiocarcinoma, the mean serum CA19-9 concentration was decreased from a preoperative level of 286.41+/-4.36 KU/L(-1) to a postoperative level of 62.01+/ 17.43 KU/L(-1) (P<0.001), and the mean serum CEA concentration from 39.41+/-24.35 microg/L(-1) to 28.69+/-11.03 microg/L(-1) (P<0.05). In patients with cholangiocarcinoma, however, no correlation was found between serum CEA and CA19 9 concentrations (r=0.036). CONCLUSION: These data suggest that the serum CA19-9 determination is a useful addition to the available tests for the differential diagnosis of cholangiocarcinoma. Serum CA19-9 is an effective tumor marker in diagnosing cholangiocarcinoma, deciding whether the tumor has been radically resected and monitoring effect of treatment. PMID- 14760773 TI - Extraction of protoporphyrin disodium and its inhibitory effects on HBV-DNA. AB - AIM: To explore an ideal method for extracting protoporphyrin disodium (PPN) from unanticoagulated animal blood, and to study the inhibitory effects of PPN on HBV DNA duplication and its cytotoxicity to 2.2.15 cell strain. METHODS: Protoporphyrin methyl ester and other intermediate products were prepared with protoheme separated from protein hydrolysates of coagulated animal blood, which were finally made into PPN and detected quantitatively with an ultraviolet fluorescent analyzer. Ten microg/ml, 20 microg/ml, 40 microg/ml, 80 microg/ml and 160 microg/ml of PPN-aqueous solution were added into culture medium for 2.2.15 cells respectively. Eight days later, the drug concentration in supernatant from the culture medium was detected when inhibition rate of HBeAg, cell survival rate when inhibition rate of HBeAg was 50% (ID50), and when survival cells in experimental group were 50% of those in control group (CD50), and the therapeutic index (TI) was also detected. PPN with different concentration of 10 microg/ml, 20 microg/ml, 40 microg/ml, 80 microg/ml and 160 microg/ml was respectively mixed and cultivated with HepG2 2.2.15 cell suspension, and then the inhibition of PPN against HBV-DNA was judged by PCR. RESULTS: The extract of henna crystal was identified to be PPN. When the concentrations of PPN were 160 microg/ml and 80 microg/ml, the inhibition rates of HBeAg were 89.8% and 82.4%, and the cell survival rates were 98.7% and 99.2%. CONCLUSION: It is suggested that PPN can be extracted from unanticoagulated animal blood. PPN can inhibit HBV-DNA expression and duplication in vitro, and has no cytotoxicity to liver cells. Further study and application of PPN are warranted. PMID- 14760774 TI - Mechanism of intrauterine infection of hepatitis B virus. AB - AIM: To explore the possible mechanism of intrauterine infection of hepatitis B virus (HBV). METHODS: HBV DNA was detected in vaginal secretion and amniotic fluid from 59 HBsAg-positive mothers and in venous blood of their newborns by PCR. HBsAg and HBcAg in placenta were determined by ABC immunohistochemistry. RESULTS: The rate of HBV intrauterine infection was 40.1% (24/59). HBV DNA was detected in 47.5% of amniotic fluid samples and 52.5% of vaginal secretion samples respectively. HBsAg and HBcAg were detected in placentas from HBsAg positive mothers. The concentration of the two antigens decreased from the mother's side to the fetus's side, in the following order: maternal decidual cells > trophoblastic cells > villous mesenchymal cells > villous capillary endothelial cells. However, in 4 placentas the distribution was in the reverse order. HBsAg and HBcAg were detected in amniotic epithelial cells from 32 mothers. CONCLUSION: The main route of HBV transmission from mother to fetus is transplacental, from the mother side of placenta to the fetus side. However, HBV intrauterine infection may take place through other routes. PMID- 14760775 TI - Multiple biomarkers of colorectal tumor in a differential diagnosis model: a quantitative study. AB - AIM: To evaluate the multiple biomarkers of colorectal tumor and their potential usage in early diagnosis of colorectal cancers. METHODS: Multiple biomarkers (DNA contents, AgNOR, PCNA, p53, c-erbB-2) in 10 normal colorectal mucosae, 37 colorectal adenomas and 55 colorectal cancers were analyzed quantitatively in the computed processing imaging system. Discrimination patterns were employed to evaluate the significance of single and multiple indices in diagnosis of colorectal cancers. RESULTS: The mean values of the analyzed parameters increased in order of the normal mucosa, adenoma and adenocarcinoma, and this tendency reflected the progression of colorectal malignancy. The parameters including DNA index, positive rates, densities of AgNOR, c-erbB-2, and p53, shape and density of nucleus were relatively valuable for diagnoses. Then a diagnostic discrimination model was established. The samples were confirmed with the model, the sensitivity rates in cancer group and adenoma group were 96.36% and 89.19%, respectively. The value of proliferating cell nuclear antigen (PCNA) in early diagnosis of colorectal cancers was uncertain. CONCLUSION: The quantitative evaluation of some parameters for colorectal tumor can provide reproducible data for differential diagnosis. The established diagnostic discrimination model may be of clinicopathological value, and can make the early diagnosis of colorectal cancer possible. PMID- 14760776 TI - Effect of antisense oligodeoxynucleotide of telomerase RNA on telomerase activity and cell apoptosis in human colon cancer. AB - AIM: To explore the effect of antisense oligodeoxynucleotide (As-ODN) of telomerase RNA on telomerase activity and cell apoptosis in human colon cancer. METHODS: As-ODN was transfected into SW480 cells by liposomal transfection reagent. Telomerase activity of SW480 cells was examined by telomeric repeat amplification protocol (TRAP) and enzyme-linked immunosorbent assay (ELISA). Apoptosis was analyzed by morphology and flow cytometry. RESULTS: The telomerase activity in SW480 cells transfected with 1.0 micromol/L of As-ODN for 2-5 days, was significantly decreased in a time-dependent manner, and the cells underwent apoptosis. The missense ODN (Ms-ODN) and the control group transfected with SW480 cells did not show these changes. CONCLUSION: As-ODN can specifically inhibit the telomerase activity of SW480 cells and induce apoptosis. PMID- 14760777 TI - Clinical and epidemiological features of patients with clonorchiasis. AB - AIM: To study the clinical and epidemiological features of patients with clonorchiasis so as to provide scientific evidences for the diagnosis and prevention of clonorchiasis. METHODS: Stools from 282 subjects suspected of having clonorchiasis were examined for helminth eggs with modified Kato's thick smear and sedimentation methods, and their sera were tested for HAV-DNA, HBV-DNA, HCV-RNA, HDV-RNA and HEV-RNA with polymerase chain reaction (PCR). Clinical symptoms of patients with clonorchiasis only were analyzed, and their blood samples were tested for circulating antigen (CAg) with Dot-ELISA, eosinophilic granulocyte count, and alanine aminotransferase (ALT). Meanwhile, they were asked to provide data of occupation, eating habit, hygienic habit and knowledge of clonorchiasis. In addition, the ecosystem of the environment in epidemic areas was surveyed. RESULTS: Among the 282 patients, 61 (21.43%) were infected with clonorchis sinensis only, 97 (34.64%) were co-infected with clonorchis sinensis and other pathogens, 92 (32.86%) were infected with hepatitis virus only and 31 (11.07%) neither with clonorchis sinensis nor hepatitis virus. Among the 61 patients with clonorchiasis only, there were 14 (22.95%) subjects with discomfort over hepatic region or epigasfrium, 12 (19.67%) with general malaise or discomfort and inertia in total body, 6 (9.84%) with anorexia, indigestion and nausea, 4 (6.56%) with fever, dizziness and headache (6.56%), and 25 (40.98%) without any symptoms; sixty one (100%) with CAg (+), 98.33% (59/60) with eosinophilic granulocytes increased and 65.00% (39/60) with ALT increased. B-mode ultrasonography revealed 61 cases with dilated and thickened walls of intrahepatic bile duct, and blurred patchy echo acoustic image in liver. Twenty six cases had stones in the bile duct, 39 cases had slightly enlarged liver with diffuse coarse spots in liver parenchyma. Twenty cases had enlarged gallbladder with thickened coarse wall and image of floating plagues, 9 cases had slightly enlarged spleen. By analysis of epidemiological data, we found that the ecologic environment was favorable for the epidemiology of clonorchiasis. Most patients with clonorchiasis were lack of knowledge about the disease. Their living environment, hygienic habits, eating habits and their occupations were the related factors that caused the prevalence of the disease. CONCLUSION: The clinical symptoms of clonorchiasis are non-specific, and the main evidences for diagnosis of clonorchiasis should be provided by etiologic examination, B-mode ultrasonography and clinical history. The infection of clonorchis sinensis is related to occupations, bad eating habits and lack of knowledge about prevention of the disease. PMID- 14760778 TI - Effects of Cd(+2), Cu(+2), Ba(+2) and Co(+2) ions on Entamoeba histolytica cysts. AB - AIM: The effects of cobalt, copper, cadmium and barium ions on the cysts of Entamoeba histolytica (E. histolytica), an amebic dysentery agent, cultured in Robinson medium were investigated. METHODS: E. histolytica cysts and trophozoites isolated from a patient with amebiasis were cultivated in the medium, incubated at 37 degrees for a period of 4 days and 40 x 10(4)/ml amebic cysts were then transferred to a fresh medium. At the second stage, 0.05, 0.1 and 0.2 mM of selected metal ions were added to the medium, and the effects of these ions on parasitic reproduction compared with the control group were observed. RESULTS: It was determined that the number of living parasites in all the groups containing metal ions decreased significantly starting from 30 minutes (P<0.01). CuCl2 showed the highest lethal effect on E. histolytica cysts, whereas the lowest lethal effect was observed with CoCl2. It was also seen that the number of living cells was decreased as the ion concentration and exposure time were increased, and that there were no living parasites in the medium at the end of 24 h (P<0.01). CONCLUSION: It may be stated that the effect of ever-increasing contamination of the environment with metal waste materials on parasites should be investigated further. PMID- 14760779 TI - Incidence of toxoplasmosis in patients with cirrhosis. AB - AIM: It is known that toxoplasmosis rarely leads to various liver pathologies, most common of which is granulomatose hepatitis in patients having normal immune systems. Patients who have cirrhosis of the liver are subject to a variety of cellular as well as humoral immunity disorders. Therefore, it may be considered that toxoplasmosis can cause more frequent and more severe diseases in patients with cirrhosis and is capable of changing the course of the disease. The aim of this study was to investigate the frequency of toxoplasmosis in patients with cirrhosis. METHODS: Serum samples were taken from 108 patients with cirrhosis under observation in the Hepatology Polyclinic of the Gastroenterology Clinic, and a control group made up of 50 healthy blood donors. IFAT and ELISA methods were used to investigate the IgG and IgM antibodies, which had developed from these sera. RESULTS: Toxoplasma IgG and IgM antibody positivity was found in 74 (68.5%) of the 108 cirrhotic patients and 24 (48%) of the 50 people in the control group. The difference between them was significant (P<0.05). CONCLUSION: In conclusion, it was found that the toxoplasma sero-prevalence in the cirrhotic patients in this study was higher. Cirrhotic patients are likely to form a toxoplasma risk group. More detailed studies are needed on this subject. PMID- 14760780 TI - Probiotics inhibit TNF-alpha-induced interleukin-8 secretion of HT29 cells. AB - AIM: To study the effect of probiotics on interleukin-8 secretion in intestinal epithelia when stimulated by proinflammatory cytokines. METHODS: Colonic adenocarcinoma HT29 cells were cultured and divided into four groups: control, TNF-alpha (group T in short), bifidobacterium (group B), lactobacillus (group L). B. Longum and L. bulgaricus were suspended in culture medium with a concentration of 1 x 10(8) cfu/ml and added into 24 wells respectively. One hour later TNF alpha (10 ng/ml) was added into each well of groups T, B, L. The supernatants were collected and measured for IL-8 after 3 hours, nuclear factor-kappaB (NF kappaB) p65 was also examined by Western blotting. RESULTS: There was less interleukin-8 secretion in HT29 cells when preincubated with B. Longum or L. bulgaricus compared with group T. Less p65 appeared in nuclei in groups B and L compared with group T, as detected by Western blot. CONCLUSION: Probiotics can suppress interleukin-8 secretion in intestinal epithelia when stimulated by proinflammatory cytokines, which is most likely mediated by NF-kappaB. PMID- 14760781 TI - Dysentery caused by Balantidium coli in a patient with non-Hodgkin's lymphoma from Turkey. AB - Balantidium coli is the only parasitic ciliate of man. It is a flattened oval organism covered with cilia, and a gullet at the anterior end. It is infrequently pathogenic for man, although epidemic buds in tropical zones have been described. The infection fundamentally affects the colon and causes variable clinic pictures, from asymptomatic to serious dysenteric forms. We present a case of parasitologically diagnosed as causes of diarrhea in a patient with non-Hodgkin's lymphoma from Turkey. In order to find out the causative etiologic agent of diarrhea, stool samples were examined by native, lugol and flotation methods and we detected moving trophozoites, which were approximately 60 microm long and 35 microm wide. These bodies were diagnosed as Balantidium coli. This case underlines that Balantidium coli should also be considered as a possible pathogen in immunocompromised patients with diarrhea. PMID- 14760782 TI - Unusual manifestations of gastric inflammatory fibroid polyp in a child. AB - AIM: Inflammatory fibroid polyp (IFP) is a rare benign lesion that may occur throughout the digestive tract. IFP is more commonly found in the antrum of the stomach in particular. It mostly affects adults at the average age of 60 years. These polyps are able to cause abdominal pain, gastrointestinal bleeding, intestinal obstruction or intussusception. In this paper we report a case of gastric IFP with unusual presenting features. METHODS: A child with gastric IFP was described and the literature was reviewed. RESULTS: A 4-year-old girl presented with fever for 2 months, arthralgia of knees and ankles, iron deficiency anemia, and hypoalbuminemia. Her stool examination was positive for occult blood. The upper gastrointestinal study demonstrated a large lobulated mass at the upper part of gastric body. Partial gastrectomy en bloc with this 5 cm x 8 cm mass was subsequently performed. Pathological examination was consistent with IFP. Following the mass excision, her fever abruptly declined and disappeared together with anemia and arthralgia. She remained asymptomatic and the abdominal ultrasonography performed at the 24-month follow-up demonstrated no recurrence of the tumor. CONCLUSION: The etiopathogenesis of IFP still remains unclear. The presence of IFP throughout the gastrointestinal tract and its variable clinical appearances make it difficult to diagnose. The inflammatory symptoms found in this patient support the hypothesis of inflammatory benign lesions of IFP. PMID- 14760783 TI - Translating research into practice: acetaminophen in osteoarthritis revisited. PMID- 14760784 TI - Rheumatological creativity and leflunomide. PMID- 14760785 TI - Trials are short, disease long: measuring drug utility beyond clinical trials. PMID- 14760786 TI - The effect of oral contraceptives and estrogen replacement therapy on the risk of rheumatoid arthritis: a population based study. AB - OBJECTIVE: Epidemiologic evidence for a protective effect of exogenous female sex hormones on the development of rheumatoid arthritis (RA) is contradictory. We examined whether exposure to either oral contraceptives (OC) or postmenopausal estrogen replacement therapy (ERT) is associated with the development of RA in women. METHODS: We separately examined the relationship between use of OC and ERT on the risk of RA in a population based case-control study. Case patients, including all female residents of Rochester, Minnesota, > or = 18 years of age, who first fulfilled 1987 American College of Rheumatology criteria for RA between 1955 and 1994 (n = 445), were compared with age matched female controls from the community. Multivariable conditional logistic regression models were used to determine whether OC or ERT exposure had an effect on RA development after controlling for potential confounders. RESULTS: We observed an inverse association between ever-use of OC and the risk of RA, which persisted after adjusting for potential confounders in multivariate analyses (OR 0.56, 95% CI 0.34, 0.92). Earlier calendar-year of first exposure to OC was associated with lower OR for RA. We found no evidence of a significant association of ERT with RA risk (adjusted OR 1.11, 95% CI 0.69, 1.78). CONCLUSION: Exposure to OC, but not ERT, significantly reduces the risk of development of RA. The risk of developing RA is lower when OC exposure occurred in earlier years, which suggests that the higher doses of estrogens and progestins contained in earlier OC preparations may have a stronger protective effect against developing RA. While this protective effect is strong, it only explains a small portion of the observed decrease in RA incidence over the past few decades because the proportion of Rochester women exposed to OC is quite small. PMID- 14760787 TI - The influence of sex on rheumatoid arthritis: a prospective study of onset and outcome after 2 years. AB - OBJECTIVE: This prospective study analyzed influence of patient's sex on early rheumatoid arthritis (RA) within one year of disease onset and after 2 years' followup. METHODS: A total of 844 consecutive patients, 538 women, with RA of less than 12 months were studied. Standardized clinical and radiographic assessments were performed at study entry and after 2 years. The association of several variables at study entry with the outcome variables Disease Activity Score (DAS28), functional disability measured by the Health Assessment Questionnaire (HAQ), and, in 390 patients, Larsen score at the 2-year followup were analyzed in men and women separately. RESULTS: At study entry the women were younger compared with the men and the sexes showed different age distributions. The women had higher DAS28 and HAQ scores. However, women below 50 years of age at study entry had milder disease than older women and close to that of men. At 2 year followup the women still had higher DAS28 and HAQ scores compared to men, who had achieved remission in a higher frequency. Larsen score showed no sex difference either at study entry or after 2 years. Presence of rheumatoid factor (RF) was associated with lower age at study entry and higher DAS28 at followup in men only. Higher DAS28 and HAQ scores at entry were more strongly correlated with severe disease at followup in women than in men. Presence of the "shared epitope" was not associated with age or the outcome variables DAS28 and Larsen score in either sex. CONCLUSION: The disease phenotype in early RA was significantly different between men and women, particularly concerning age, disease activity, and functional capacity. There were differences between the sexes concerning early disease characteristics associated with outcome at 2 years of followup. PMID- 14760788 TI - A web-compatible instrument for measuring self-reported disease activity in arthritis. AB - OBJECTIVE: To describe a Web-based computer health assessment survey for patients with rheumatoid arthritis (RA) and to evaluate the survey in comparison with current paper versions. METHODS: Utilizing data from a study on RA, we compared results from 43 patients attending a university-based clinic who were each given a paper and a demonstration computer version of a patient self-assessment questionnaire including multiple-choice questions from a multi-dimensional Health Assessment Questionnaire (HAQ); visual analog scales (VAS) for pain, fatigue, and global disease severity; and a tender and swollen joint count reporting tool. Patients were given optional followup surveys to determine their opinion of the computer program. RESULTS: High correlations (intraclass correlation coefficient > 0.9) were seen across methods for the 10-item HAQ and psychological distress scores and the VAS scores for pain and global disease severity. Moderate correlation was observed for the self-efficacy scores, the VAS scores for fatigue, and tender joint counts. The data also revealed a small shift in the mean scores for the HAQ and self-efficacy questions, with patients reporting slightly higher scores on the computer instrument. Overall, patient opinions of the uniquely designed joint count tool were good, with 71% of responding patients answering favorably. CONCLUSION: Web-based computer versions of patient self assessment surveys in RA are comparable to paper versions, and their use in clinics or over the Internet could dramatically facilitate the ability of physicians to monitor patients' health. PMID- 14760789 TI - Patient questionnaires and formal education level as prospective predictors of mortality over 10 years in 97% of 1416 patients with rheumatoid arthritis from 15 United States private practices. AB - OBJECTIVE: To prospectively analyze patient questionnaire scores concerning functional disability as well as formal education level as potential predictors of premature mortality over 10 years in 1416 patients with rheumatoid arthritis (RA) from 15 private practice rheumatology settings in 11 diverse cities in the United States. METHODS: At baseline in 1985 and periodically over 10 years, patients completed mailed self-report multidimensional health assessment questionnaires (MDHAQ) that included functional disability scores, formal education level, and other demographic and clinical data. Vital status was determined 10 years after baseline. Potential predictors of 10 year mortality were analyzed using descriptive statistics and Cox proportional hazards models. RESULTS: Vital status was accounted for in 1378 patients, 97.3% of the cohort. The standard mortality ratio was 1.6, similar to most reported series of patients with RA, as 401 patients died versus 251 expected over 10 years. Evidence of "dose-response" relations was seen for age, formal education level, functional disability scores, and helplessness scores as predictors of mortality. In Cox proportional hazards models, age, sex, formal education level, functional disability, and helplessness scores remained significant independent predictors of 10 year mortality. CONCLUSION: Functional disability and low formal education level are significant predictors of premature mortality in people with RA under care in US private practice settings, as in most reported cohorts of patients with RA. This study shows that it is possible to account for more than 95% of patients over 10 years using mailed questionnaires to monitor patient status. PMID- 14760790 TI - Rheumatoid arthritis treatment with weekly leflunomide: an open-label study. AB - OBJECTIVE: To assess the safety and effectiveness of leflunomide (LNF) using 100 mg/week in patients with rheumatoid arthritis (RA). METHODS: Patients who were clinically active using the American College of Rheumatology criteria for RA were enrolled. They received a loading dose of 100 mg of LFN for 3 days, followed by 100 mg of LFN weekly. Efficacy and adverse events (AE) were recorded. RESULTS: Fifty patients were enrolled; 46 (93.6%) were women with a mean age of 45.6 years (range: 24 to 83). Disease duration was 3.7 years (range: 0.5 to 12). Twenty patients (40.8%) had previously taken disease modifying antirheumatic drugs. Outcomes achieved after 24 weeks of treatment were as follows: ACR20 (74%), ACR50 (64%), and ACR70 (28%). Five patients were withdrawn due to AE: 2 due to urticaria, 2 patients had elevated liver enzymes, and one had thrombocytopenia. Six patients (12%) were lost to followup. No severe AE were seen. CONCLUSION: The results in our preliminary report indicate that using a 100 mg/week dose achieves a similar benefit to the LFN 20 mg/day treatment, and there were no severe AE. In addition, a single LFN weekly dose has better treatment compliance. A secondary important benefit is the reduction of the monthly cost of medication. Comparative and blind trials are necessary in order to confirm longterm improvement and benefits on this regimen. PMID- 14760791 TI - Effect of repeated infliximab therapy on serum matrix metalloproteinases and tissue inhibitors of metalloproteinases in patients with rheumatoid arthritis. AB - OBJECTIVE: Matrix metalloproteinases (MMP) are involved in the articular tissue destruction processes in the pathogenesis of rheumatoid arthritis (RA). We investigated the effects of multiple infusions of infliximab, a chimeric anti tumor necrosis factor-a (anti-TNF-a) antibody, on concentrations of serum MMP and tissue inhibitors of metalloproteinases (TIMP) in patients with active RA. METHODS: Patients with RA were scheduled to receive 9 infusions of infliximab (3 mg/kg) at Weeks 0, 2, 6, and every 8 weeks thereafter. The therapy was combined with methotrexate (MTX) (7.5-20 mg/week). Serum concentrations of interstitial collagenase (MMP-1), stromelysin-1 (MMP-3), gelatinase B (MMP-9), TIMP-1, and TIMP-2 were measured by ELISA prior to infusion at Weeks 0, 2, 6, 14, 38, and 62. RESULTS: The initial infusion of infliximab downregulated serum levels of MMP-1 (p < 0.001), MMP-3 (p < 0.001), MMP-9 (p < 0.001), TIMP-1 (p < 0.01), and TIMP-2 (p < 0.05). The second drug administration caused even more remarkable reduction of measured MMP (p < 0.001 in all cases) but not of TIMP levels. These changes were accompanied by decreased ratios of measured MMP to TIMP. Further infliximab therapy also significantly suppressed serum MMP levels, but was less effective. Before the first infliximab infusion serum concentrations of MMP and TIMP correlated with markers of RA activity such as the Disease Activity Score and C reactive protein levels. After further drug administrations such associations, although less significant, were also noted. CONCLUSION: Anti-TNF-a antibody therapy combined with MTX resulted in rapid clinical improvement and reduced serum MMP concentrations in patients with RA. Further infusions of infliximab maintained the decrease of MMP, although to a lesser extent than the first and second doses. PMID- 14760792 TI - Efficacy and safety of tacrolimus (FK506) in treatment of rheumatoid arthritis: a randomized, double blind, placebo controlled dose-finding study. AB - OBJECTIVE: To evaluate the efficacy and safety of tacrolimus (FK506) in patients with active rheumatoid arthritis (RA) exhibiting resistance to disease modifying antirheumatic drug (DMARD) therapy, and to determine the optimal dosage. METHODS: A total of 212 patients with DMARD-resistant RA were enrolled in this double blind, multicenter, randomized, placebo controlled study and allocated to 3 groups. Patients were administered tacrolimus at a dosage of 1.5 mg/day (68 patients) or 3 mg/day (70 patients), or placebo (74 patients), for 16 weeks. They were allowed to continue taking prednisolone (< or = 5 mg/day) and/or one nonsteroidal antiinflammatory drug (NSAID) during the study. Clinical assessment was based on the American College of Rheumatology (ACR) 20% criteria. RESULTS: ACR 20% response rates were higher in both tacrolimus groups (3 mg: 48.3%; 1.5 mg: 24.6%) than in the placebo group (14.1%), with the rate in the 3 mg group significantly higher. There were no significant differences between the tacrolimus groups and placebo group in the incidence of adverse events. The main adverse events in the tacrolimus groups, especially in the 3 mg group, were renal function abnormalities and gastrointestinal symptoms. However, no significant differences were observed among the 3 groups in the incidence of any adverse event except decrease in serum Mg level. CONCLUSION: Our findings demonstrate excellent dose-dependent efficacy of tacrolimus in patients with DMARD-resistant RA and strongly suggest the usefulness of tacrolimus for treatment of RA. The optimal dosage appears to be 3 mg/day in terms of efficacy and safety. PMID- 14760793 TI - Autoimmune response to proteasome activator 28alpha in patients with connective tissue diseases. AB - OBJECTIVE: To determine the autoimmune response against proteasome activator 28alpha (PA28alpha) in patients with various connective tissue diseases, and to compare the immunoreactivity between anti-PA28alpha and anti-Ki antibodies. METHODS: Serum samples were obtained from 219 patients with various connective tissue diseases. cDNA encoding full-length human PA28alpha and Ki were produced by polymerase chain reaction. Antigens were expressed as glutathione S transferase (GST) fusion proteins. The immunoreactivity of serum for PA28alpha and Ki was studied by Western blotting. An inhibition test was performed by ELISA using purified Ki antigen. RESULTS: Anti-PA28alpha> antibody was detected in serum from 23% of patients with systemic lupus erythematosus (SLE) and 24% with Sjogren's syndrome (SS). These rates were significantly higher than those for the other rheumatic diseases. Since both PA28alpha and Ki are elements of the PA28 complex and their amino acid sequences share 40.2% homology, immunoreactivity to PA28alpha was studied further. Among 27 anti-Ki positive serum samples, 13 samples (48%) also reacted with PA28alpha, suggesting a relationship between anti PA28alpha and anti-Ki antibodies. To investigate whether this finding was due to the presence of cross-reacting epitopes for PA28alpha and Ki antigens, an inhibition test was performed by ELISA. The reactivity to purified Ki antigen was not inhibited by preincubation with recombinant PA28alpha. CONCLUSION: Detection of anti-PA28alpha antibody was significantly higher in serum from patients with SLE and SS. The relationship between anti-PA28alpha and Ki antibodies suggests the importance of an antigen-driven system in the induction of an autoimmune response to PA28 complex. PMID- 14760794 TI - Effects of disease activity, pain, and distress on activity limitations in patients with systemic lupus erythematosus. AB - OBJECTIVE: To evaluate the extent to which disease activity, pain, and psychological distress predict activity limitations in persons with systemic lupus erythematosus (SLE). METHODS: A sample of 93 persons with SLE completed medical and psychosocial evaluations at one study visit. Sets of measures were chosen to represent constructs of Disease Activity, Pain, Distress, and Activity Limitations. Confirmatory factor analysis was conducted to determine if the measures fit the intended constructs, and structural equation modeling was used to evaluate the direct effects of Disease Activity, Pain, and Distress on Activity Limitations, as well as indirect effects of Disease Activity as mediated by Pain and Distress, and indirect effects of Pain as mediated by Distress. RESULTS: The confirmatory factor analysis indicated a good fit of the measures to their intended constructs. The overall model predicting Activity Limitations based on Disease Activity, Pain, and Distress accounted for 68% (p < 0.001) of the variance in perceived Activity Limitations. Severity of Pain was the only construct that was directly associated with Activity Limitations (r = 0.70, p < 0.001). The effects of Disease Activity on Activity Limitations were primarily indirect via its influence on Pain. Distress was not significantly associated with Activity Limitations. When Distress was trimmed from the model, the remaining constructs accounted for 66% of the variance in Activity Limitations (p < 0.001). CONCLUSION: Disease Activity and Pain accounted for a substantial proportion of the variance in Activity Limitations. Pain Severity was the strongest predictor of Activity Limitations. This study highlights the importance of adequate pain management for maintaining quality of life in persons with SLE. PMID- 14760795 TI - Macrophage migration inhibitory factor in systemic lupus erythematosus. AB - OBJECTIVE: To examine associations between serum macrophage migration inhibitory factor (MIF) and disease-related variables and corticosteroid use in patients with systemic lupus erythematosus (SLE). METHODS: Serum MIF concentration was measured by ELISA in 90 female patients with SLE and 279 healthy controls. Univariate and multivariate regression analyses were used to examine the associations between serum MIF concentration and disease-related indices of SLE and corticosteroid use. RESULTS: Serum MIF concentrations were positively associated with SLE disease damage (SLICC/ACR index), and indices of disease damage were greater in SLE patients with serum MIF concentrations above the normal median value. Serum MIF concentration was also observed to be significantly greater in patients with SLICC/ACR damage index (DI) scores >/= 3. Serum MIF was also positively associated with current corticosteroid dose. Significantly higher SLICC/ACR DI scores were observed in patients with values of serum MIF above the normal median, and this remained significant after adjusting for corticosteroid dose. Serum MIF concentration was also predictive of SLICC/ACR index after 3 years of followup, but this association was partly confounded by corticosteroid dose. Serum MIF was also negatively associated with serum creatinine concentration, independent of disease damage and corticosteroid dose. CONCLUSION: MIF is overexpressed in patients with SLE. While this can be partly explained by corticosteroid use, there is evidence of an association between MIF and lupus-related disease damage. PMID- 14760796 TI - Close association of herpes zoster reactivation and systemic lupus erythematosus (SLE) diagnosis: case-control study of patients with SLE or noninflammatory musculoskeletal disorders. AB - OBJECTIVE: To investigate the prevalence of infections, particularly the frequency of shingles and the timing of varicella zoster virus (VZV) reactivation, and antibiotic use, vaccinations, and joint trauma prior to and at diagnosis of systemic lupus erythematosus (SLE). METHODS: We sent questionnaires to patients with SLE (n = 93) and controls with noninflammatory musculoskeletal disorders (MSK; n = 353) including osteoarthritis, fibromyalgia, and tendonitis. We matched SLE patients to controls for sex (up to 1:3). RESULTS: The response rate in SLE was 66% and in controls 69% (p < 0.53). Four of 61 SLE patients and 12 of 173 controls were men. The mean disease duration in the SLE group was 8 +/- 1 years compared to 10 +/- 1 years in controls (p < 0.23). SLE patients were significantly younger than controls (mean age of SLE patients 49 +/- 2 vs 57 +/- 1 years for controls; p < 0.0004), and results were adjusted for age. A significantly higher proportion of SLE participants had a history of VZV (shingles) (19% vs 7%, respectively; OR 2.98, p < 0.003), whereas rubella was reported less in SLE (23% vs 42%; OR 0.43, p < 0.03). VZV infections were clustered just prior to or after diagnosis in SLE but were more widely spaced temporally in the controls (1 +/- 4.5 years after the diagnosis of SLE vs -14.7 +/- 4 years before the diagnosis of noninflammatory MSK disorder; p < 0.003). Diagnosis of shingles was observed in 6 of 11 SLE patients within +/- 2 years of SLE diagnosis, whereas only 2 of 15 controls had shingles within +/- 2 years of diagnosis (OR 7.2, p < 0.03). Only 2 patients with SLE were taking immunosuppressive drugs or steroids at time of shingles, so immunosuppressive therapy was not usually concomitant at time of VZV reactivation. Common infections (respiratory, urinary tract, ear, and eye) in the SLE group exceeded controls, but not significantly (23% vs 9%; OR 2.98, p < 0.06) and SLE patients were more likely to have been vaccinated since 18 years of age with any type of vaccine (69% vs 51%; OR 2.21, p < 0.04). SLE patients were less likely than controls to report joint trauma within one year prior to their diagnosis (25% vs 40%; OR 0.49, p < 0.04). There were no differences with respect to streptococcal throat infection (p < 0.96), diarrhea/vomiting (p < 0.84), rash with fever (p < 0.07), parvovirus infection (p < 0.16), infection after surgery (p < 0.58), respiratory tract infection (p < 0.71), or ear (p < 0.09) and eye infection (p < 0.68) one year prior to diagnosis. A higher proportion of SLE patients had a history of urinary tract infections (46% vs 25%), but this was not significant (p < 0.17), nor was it significant one year prior to diagnosis (p < 0.63). Overall, the likelihood of having any infection one year prior to diagnosis was not significantly higher in the SLE group (p < 0.56). There were no differences one year prior to diagnosis in travel history (p < 0.69), hospitalizations (p < 0.47), use of antibiotics (p < 0.54), history of rheumatic fever, positive TB skin test, or hepatitis A, B or C infection. CONCLUSION: Varicella reactivation as shingles is increased in patients with SLE and clusters around diagnosis. Vaccinations are increased in those with SLE compared to controls. Common infections are not significantly increased in SLE patients prior to onset of symptoms. We cannot determine if VZV infections are causally associated with SLE in some people, are from an abnormal immune system response due to the lupus itself or from the use of steroids or other immunosuppressive drugs to control the disease, or are spurious. PMID- 14760797 TI - Transverse myelitis as the first manifestation of systemic lupus erythematosus or lupus-like disease: good functional outcome and relevance of antiphospholipid antibodies. AB - OBJECTIVE: Transverse myelitis (TM) is a rare complication of systemic lupus erythematosus (SLE). Although usually a late manifestation of SLE, it can occur at presentation. We investigated the clinical presentation, treatment and outcome of 15 patients with TM as the presenting manifestation of SLE or lupus-like disease. METHODS: All patients received corticosteroids, while 13 also received immunosuppressive therapy. Five patients were fully anticoagulated with warfarin. RESULTS: A sensory level with spastic lower limb weakness and sphincter disturbance was the most common presentation: 14/15 patients had a thoracic or cervical sensory level. Cerebrospinal fluid examination showed high protein concentrations in 3 patients and oligoclonal bands in 8. Eleven of the 15 (73%) had antiphospholipid antibodies (aPL). Of the 15 patients, 3 had complete resolution of the symptoms, 6 had good functional improvements, 5 had good to fair outcome with some functional deficit, and one patient who received corticosteroids alone later died from pneumonia. CONCLUSION: We describe 15 patients with TM as the presenting manifestation of SLE or lupus-like disease with a high prevalence of aPL. Our data support the view that early diagnosis and immunosuppressive therapy may be superior to corticosteroids alone in improving functional outcome. In those patients with aPL, antiplatelet agents and/or warfarin should also be considered. PMID- 14760799 TI - Interleukin 1beta gene polymorphism association with severe renal manifestations and renal sequelae in Henoch-Schonlein purpura. AB - OBJECTIVE: To assess the influence of the interleukin (IL)-1beta gene (-511 C/T) in the incidence of Henoch-Schonlein purpura (HSP) and determine its possible implication in severe systemic complications of HSP, in particular severe renal involvement and permanent renal dysfunction (renal sequelae). METHODS: Patients from Northwest Spain with primary cutaneous vasculitis classified as HSP according to proposed criteria were studied. All patients were required to have had at least 2 years' followup. Patients and ethnically matched controls were genotyped for IL-1beta gene (-511 C/T) polymorphism by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP). RESULTS: Forty-nine Caucasian patients (38 of them younger than 21 years) who fulfilled classification criteria for HSP and 148 controls were examined. No allele or genotype differences between the whole group of HSP and controls were observed. However, all 5 patients who developed severe nephropathy during the course of disease carried the rare T allele compared with only 16 of the remaining 44 patients (pcorr = 0.01). A significant association between carriage of the 511(IL-1beta) T allele and renal sequelae (pc = 0.02; OR: 3.6, 95% CI: 1.3-10.0) was also found. CONCLUSION: In unselected patients with cutaneous vasculitis who fulfill classification criteria for HSP, carriage of IL-1beta (-511) T allele appears to influence severity of renal involvement. PMID- 14760798 TI - Nailfold capillary microscopy can suggest pulmonary disease activity in systemic sclerosis. AB - OBJECTIVE: To evaluate the association of capillaroscopic alterations with pulmonary disease activity in systemic sclerosis (SSc). METHODS: Ninety-one patients with SSc were studied by means of interview, physical examination, nailfold capillary microscopy (NCM), serology, pulmonary function tests, esophageal transit scintigraphy, Doppler echocardiography, and pulmonary high resolution computed tomography (HRCT). Pulmonary disease activity was diagnosed by the observation of ground-glass opacities on pulmonary HRCT. Capillary loss on NCM was evaluated using the avascular score: patients with mean score > or = 1 or mean number of megacapillaries per finger > or = 1 were considered to have severe capillaroscopic alterations. RESULTS: Patients with higher skin scores, longer disease duration, signs of peripheral ischemia, esophageal dysfunction, antitopoisomerase I antibodies, and ground-glass opacities had higher mean avascular scores (p < or = 0.05 in all tests). The association between ground glass opacities and higher avascular scores was particularly strong in patients with disease duration < or = 5 years. Among these patients, ground-glass opacities were present in 14 of 19 patients with severe NCM alterations, but were absent in all patients (n = 8) with mild or no NCM alterations (p < 0.001). ROC curves confirmed the ability of NCM to discriminate between patients with and without ground-glass opacities among those with disease duration < or = 5 years. However, NCM could not predict the presence of reduced pulmonary diffusing capacity. CONCLUSION: The severity of NCM abnormalities is associated with lung disease activity in SSc, particularly when the disease duration is relatively short. PMID- 14760800 TI - Lack of association between endothelial nitric oxide synthase polymorphisms and Henoch-Schonlein purpura. AB - OBJECTIVE: To assess the influence of endothelial nitric oxide synthase (eNOS) polymorphisms in the susceptibility and clinical expression of patients with cutaneous vasculitis fulfilling classification criteria for Henoch-Schonlein purpura (HSP). METHODS: Fifty patients from Northwest Spain with primary cutaneous vasculitis classified as HSP were studied. Patients and ethnically matched controls (n = 117) were genotyped by polymerase chain reaction techniques for a variable-number tandem-repeat polymorphism in intron 4, a T/C polymorphism at position -786 in the promoter region, and a polymorphism in exon 7 (298Glu/Asp or 5557G/T) of the eNOS gene. RESULTS: No differences in allele or genotype frequencies for any of the individual eNOS polymorphisms were observed between patients fulfilling HSP classification criteria and controls, or when patients were stratified for the presence of nephritis or joint or gastrointestinal manifestations. In the HSP group no linkage disequilibrium between these polymorphisms was found. No significant difference in haplotype frequencies was observed between patients and controls. CONCLUSION: Our results do not support a role for these polymorphisms in the susceptibility to HSP. PMID- 14760801 TI - Galanin in adjuvant arthritis in the rat. AB - OBJECTIVE: To study the concentration changes of galanin in the ankles and spinal cord and to detect the distribution of galanin in different tissues in arthritic rats. METHODS: Adjuvant arthritis was induced by intradermal injection of Mycobacterium butyricum in Freund's incomplete adjuvant at the base of the tail. The concentrations of galanin were measured by radioimmunoassay (RIA) and the distributions of galanin were detected by immunoelectron microscopy (iEM). RESULTS: Measurements were taken on Day 28 after injection. RIA results showed that the concentration of galanin was significantly lower in the ankles and spinal cords of rats with adjuvant arthritis compared to controls. Our iEM results showed a heterogeneous distribution of galanin labelling in different cells and tissue compartments. In arthritic rats, we observed decreased galanin labelling in the sciatic nerve and in macrophage-like cells in the synovial membrane and increased labelling in monocyte lineage cells, polymorphonucleated lineage cells in the bone marrow, fibroblasts in the periosteum, osteoclasts and osteocytes, and lower labelling in osteoblasts compared to controls. CONCLUSION: Galanin is involved in the response to inflammation in adjuvant arthritis and might play a role in the regulation of inflammation and nociception. These findings are in accordance with a biological role of galanin in the development of inflammatory arthritis. PMID- 14760802 TI - A randomized comparative study of short term response to blind injection versus sonographic-guided injection of local corticosteroids in patients with painful shoulder. AB - OBJECTIVE: Local corticosteroid injections, commonly accepted by rheumatologists to be effective treating painful shoulder, have shown controversial results. High frequency ultrasonography is an accurate and safe imaging modality for guiding musculoskeletal injections. We prospectively compared the short term response to randomized blind injection versus sonographic-guided injection of local corticosteroid in patients with painful shoulder. METHODS: We studied 41 consecutive patients with painful shoulder. Patients with previous trauma or chronic inflammatory arthritis were excluded. No patient had received previous physiotherapy or local steroid injection in the shoulder. Patients were randomized to receive either a blind subacromial injection of 20 mg triamcinolone (Group 1, n = 20) or a sonographic guided injection of 20 mg triamcinolone (Group 2, n = 21) by the same rheumatologist blinded to the clinical evaluation. In both groups we recorded shoulder abnormalities and the location of the steroid postinjection by ultrasound. Each patient was clinically assessed within 5 days before injection and 6 weeks after injection by another rheumatologist without knowledge of the injection technique performed. Clinical assessment included demographic and clinical data, a visual analog scale (VAS) for pain (0-100), the Shoulder Function Assessment (SFA) scale (0-70), and postinjection adverse effects. No patient received physical therapy during the followup period. Initially, demographic, clinical, and ultrasonographic findings in both groups showed no significant differences. RESULTS: Six weeks after injection, the VAS and the SFA score showed a significantly greater improvement in Group 2 compared with Group 1 (mean VAS score change 34.9 for Group 2 vs 7.1 for Group 1, p < 0.001; and mean SFA score change 15 for Group 2 vs 5.6 for Group 1, p = 0.012). One patient in Group 1 reported mild postinjection adverse effects. CONCLUSION: We suggest that sonographic-guided corticosteroid injections should be indicated, at least, in patients with poor response to previous blind injection to ensure accurate medication placement in order to improve therapeutic effectiveness. PMID- 14760803 TI - ADAMTS-1, a gene product of articular chondrocytes in vivo and in vitro, is downregulated by interleukin 1beta. AB - OBJECTIVE: Osteoarthritic (OA) cartilage degeneration and cartilage destruction in rheumatoid arthritis depends significantly on enzymatic degradation of cartilage proteoglycan aggrecan. A member of the ADAMTS family of proteases, ADAMTS-1 was described to have "aggrecanase" activity. We investigated the quantitative expression and distribution of ADAMTS-1 in healthy and OA cartilage and in cultured articular chondrocytes with and without stimulation by interleukin 1beta (IL-1beta) and insulin-like growth factor-I (IGF-I). METHODS: Conventional and online polymerase chain reaction (PCR) technology was used to determine ADAMTS-1 mRNA expression levels of ADAMTS-1. Protein was localized using immunostaining with different polyclonal antibodies. RESULTS: Conventional and online semiquantitative PCR showed significant levels of ADAMTS-1 mRNA expression in normal and OA chondrocytes in vivo and in vitro. Only a slight increase was observed in OA cartilage. After stimulation with IL-1beta a downregulation of ADAMTS-1 was observed, whereas IGF did not appear to change mRNA expression levels in vitro. The in vivo mRNA expression results were confirmed by the presence of significant protein staining with antibodies for ADAMTS-1 in normal and OA chondrocytes as well as Western blotting analysis. Whereas a significantly stronger stain was seen in normal articular cartilage in the upper zones, in OA cartilage as well the middle zone showed enhanced staining. CONCLUSION: Our results confirm the expression and presence of ADAMTS-1 in articular cartilage. However, they also point out that ADAMTS-1 appears to be constitutively expressed by adult articular chondrocytes and overall is not strongly upregulated in OA. Thus our data suggest that ADAMTS-1 is the first matrix-degrading enzyme downregulated by the catabolic factor IL-1beta in vitro. PMID- 14760804 TI - Effect of radiosynovectomy with holmium-166 ferric hydroxide macroaggregate on adult equine cartilage. AB - OBJECTIVE: To analyze the effect of radiosynovectomy with holmium-166 ferric hydroxide macroaggregate (166Ho-FHMA) on articular cartilage in 6 adult horses. METHODS: Arthritic changes and mechanical properties of articular cartilage were evaluated with arthroscopy and postmortem microscopic analyses. Glycosaminoglycan content was measured by safranin-O staining combined with digital densitometry, uronic acid analyses, and dimethylene blue binding assay. 35S-sulfate labeling and autoradiography were used to localize proteoglycan synthesis and to characterize proteoglycan structures using SDS-agarose gel electrophoresis. Northern hybridizations were performed to measure the mRNA levels for aggrecan and pro-a1(II) collagen in cartilage samples. RESULTS: Histological signs of degeneration were present in the articular cartilage of both control and radiosynovectomized equine joints. Radiosynovectomy did not aggravate degenerative changes or significantly alter the matrix glycosaminoglycan content. A slightly decreased size of proteoglycan monomers was observed 2 months after 166Ho-FHMA radiosynovectomy. Tissue analysis of extracted proteoglycans revealed lower 35S incorporation after radiosynovectomy, but corresponding changes could not be observed in aggrecan mRNA levels. Transient downregulation of pro-a1(II) collagen mRNA transcription was observed 5 days after 166Ho-FHMA radiosynovectomy. CONCLUSION: 166Ho-FHMA treatment did not markedly affect the composition or morphology of adult articular cartilage showing mild degeneration. However, minor degradation of proteoglycan monomers and transient downregulation of pro-a1(II) collagen mRNA were observed. PMID- 14760805 TI - Bone scintigraphy is not a better predictor of progression of knee osteoarthritis than Kellgren and Lawrence grade. AB - OBJECTIVE: To determine the predictive value of bone scintigraphy with respect to joint space narrowing (JSN) in patients with knee osteoarthritis (OA), based on quantitative estimates of uptake of a bone-seeking radiopharmaceutical and fluoroscopically standardized knee radiography. METHODS: Our study group included 86 obese women, 45-64 years of age, with unilateral knee OA. Uptake of technetium medronate (99mTc-MDP) in late-phase bone scans was measured at baseline in 5 regions of interest (ROI: lateral femur, lateral tibia, medial femur, medial tibia, and patellofemoral joint) and was adjusted for uptake (i.e., expressed as a ratio to uptake) in a ROI in the shaft of the tibia, which served as an internal standard. Each subject underwent a fluoroscopically standardized radiograph of the knees (semiflexed anteroposterior view) at baseline, 16, and 30 months. Magnification-corrected minimum joint space width in the medial tibiofemoral compartment was measured by digital image analysis. RESULTS: Followup was available for 79 patients (92%) at 16 months and from 73 patients (85%) at 30 months. On average, 99mTc-MDP uptake in each ROI and in the whole knee (average of 4 tibiofemoral ROI) was 170-240% of that in the tibial shaft. Uptake in the medial tibia and in the whole knee was significantly correlated with JSN at 16 and 30 months (r = 0.22-0.30, p < 0.05). However, after controlling for age, body mass index, and radiographic severity of OA, the associations between adjusted uptake and JSN were not significant. The rate of JSN in knees of patients with OA who were in the lower tertile with respect to adjusted 99mTc-MDP uptake in the medial tibia was significantly less rapid than in patients in whom uptake was in the middle and upper tertiles (0.04 mm/yr vs 0.18 mm/yr; p < 0.05). However, after controlling for overall radiographic severity at baseline, the difference in 30-month JSN in knees of patients with OA in the lower versus middle/upper tertiles was not significant. CONCLUSION: The predictive utility of bone scintigraphy is confirmed by these data. However, its practical value is considerably diminished, insofar as similarly predictive information may be obtained by routine radiographic examination, without the radiation exposure and cost of scintigraphy. PMID- 14760806 TI - A randomized, single-blind comparison of the efficacy and tolerability of hylan G F 20 and triamcinolone hexacetonide in patients with osteoarthritis of the knee. AB - OBJECTIVE: To assess prospectively the efficacy and tolerability of hylan G-F 20 (HG-F 20; Synvisc) and intraarticular triamcinolone hexacetonide (TH; Aristospan) for treatment of osteoarthritis (OA) knee pain in a 26 week, randomized, multicenter, evaluator-blind study. METHODS: Patients with OA were treated with typical regimens of HG-F 20 (n = 113) and TH (n = 102). Primary assessments were the WOMAC question A1 (pain walking on a flat surface), and a 100 mm visual analog scale (VAS) for patient and investigator overall assessments. Total WOMAC and WOMAC domain C (function) scores were also assessed. The intent-to-treat population was analyzed using a last-observation carried forward approach. RESULTS: Maximum pain relief occurred at 1-2 weeks for TH and at Week 12 for HG-F 20. At Weeks 12 and 26, HG-F 20 was significantly better than TH for the WOMAC question A1 responses (p = 0.0071 and p = 0.0129, respectively), and patient VAS (p < 0.0001 and p < 0.0001) and investigator VAS (p < 0.0300 and p = 0.0004) assessments. Similar significant (p < 0.01) results were observed at Weeks 12 and 26 for total WOMAC and domain C scores. While 15 TH-treated patients discontinued the study due to lack of efficacy, none did so with HG-F 20 treatment (p < 0.01). Both agents were well tolerated with similar adverse event profiles. CONCLUSION: Viscosupplementation with HG-F 20 resulted in a longer duration of effect than TH with a comparable tolerability profile. These data support the preferential use of HG-F 20 over TH for treatment of chronic OA knee pain. PMID- 14760809 TI - Clinical profile of rheumatic disease patients referred to a multidisciplinary pain center. AB - OBJECTIVE: Good pain control is a prerequisite for success in the management of many rheumatological diseases. However, some rheumatology patients may present challenges in terms of pain management and be subsequently referred to a specialized pain clinic. We examined the characteristics and assessed the outcome of patients with rheumatic diseases who were referred to a tertiary care pain center. METHODS: All new patients with a primary rheumatological diagnosis referred over a 9 year period to the McGill University Pain Centre were studied. Patients were identified through a computer search according to both diagnoses and symptoms. Demographic information, clinical and pain characteristics, and subsequent management and final outcome were assessed. RESULTS: Out of a total of 1120 new patients, 60 (5%) had a primary rheumatologic diagnosis to account for pain and referral. The diagnoses were as follows: fibromyalgia in 26 (43%), inflammatory arthritis 17 (28%), degenerative arthritis 9 (15%), and soft tissue rheumatism 8 (13%). The median age at presentation was 52 years and 47 (78%) were female. The median duration of pain was 5 years. The mean pain scores according to the McGill Pain Questionnaire and the visual analog scale were 27 +/- 15 and 7 +/- 2, respectively. Patients were followed a mean duration of 10.6 +/- 15 months. Seventy-two percent were assessed by a psychologist and 52% by a physiotherapist or occupational therapist. New pharmacologic treatments were prescribed for 47 (78%) patients, with 47% receiving opioids, 37% antidepressants, 12% nonsteroidal antiinflammatory drugs, 8% tranquillizers, and 18% other medications. Final outcome was described as follows: improved in 55%, no change in 43%, and worsened in 2%. CONCLUSION: Although patients with a primary rheumatologic process to account for pain constituted a small proportion of patients evaluated, improvement was considerable in over half. Further study should address the selection of patients that are most likely to benefit from referral to multidisciplinary pain centers and the longterm outcome of such interventions. PMID- 14760807 TI - Nonsteroidal antiinflammatory drugs or acetaminophen for osteoarthritis of the hip or knee? A systematic review of evidence and guidelines. AB - OBJECTIVE: The interpretation of available evidence on the relative efficacy of nonsteroidal antiinflammatory drugs (NSAID) and acetaminophen in osteoarthritis (OA) has recently been debated. This systematic review summarizes the available evidence on the efficacy of NSAID compared to acetaminophen, and compares the quality and content of clinical guidelines regarding the pharmacological treatment of OA. METHODS: Published reports of randomized controlled trials (RCT) and clinical guidelines were identified by a systematic search of bibliographic databases and relevant websites. The quality of RCT was assessed by 2 reviewers independently using a standardized checklist. Data from these RCT were used to calculate pooled differences between groups for pain and disability. The methodology of identified guidelines was appraised using the AGREE (Appraisal of Guidelines for Research and Evaluation) instrument. RESULTS: The search strategy resulted in the identification of 5 RCT. Statistical pooling of data from 3 trials with adequate methods and sufficient data presentation resulted in a pooled standardized mean difference for general pain of 0.33 (95% CI 0.15 to 0.51), indicating a small effect in favor of NSAID. Pooled estimates for other outcome measures were smaller. Three of the 9 identified guidelines satisfied more AGREE criteria than others, particularly regarding rigor of development. Stakeholder involvement, applicability, and editorial independence were poorly described in most guidelines. The content of recommendations regarding the use of NSAID or acetaminophen was fairly consistent. CONCLUSION: Acetaminophen is often effective in OA and is associated with fewer adverse reactions than NSAID. Available evidence supports the recommendations of recent guidelines to use acetaminophen as initial therapy for OA in addition to nonpharmacological interventions. Further research is needed to establish the efficacy of NSAID or acetaminophen in relevant subgroups of patients. We agree with guidelines that it is important that treatment is tailored to individual patients taking into account the severity of symptoms, previous use of acetaminophen, and the patient's knowledge, expectations, and preferences. PMID- 14760808 TI - Longer use of COX-2-specific inhibitors compared to nonspecific nonsteroidal antiinflammatory drugs: a longitudinal study of 3639 patients in community practice. AB - OBJECTIVE: To compare COX-2-specific inhibitor therapy with conventional nonspecific nonsteroidal antiinflammatory drugs (NS NSAID), and investigate the effect of demographic and disease factors on NSAID duration of use. METHODS: A total of 3639 patients with rheumatoid arthritis (RA), osteoarthritis, and fibromyalgia starting therapy of celecoxib, rofecoxib, naproxen, or ibuprofen were surveyed at 6-month intervals for up to 2.5 years. Detailed demographic and disease severity variables were also measured. Time to discontinuation, discontinuation rates, and effect of covariates were determined by Weibull parametric survival analyses, controlling for a wide variety of demographic and disease severity factors. RESULTS: The median duration of use for celecoxib, rofecoxib, naproxen, and ibuprofen was 15, 13, 10, and 10 months, respectively. Duration of use of celecoxib and rofecoxib, as measured by survival times, was significantly longer than those of naproxen and ibuprofen. The celecoxib survival time was significantly longer than the rofecoxib survival time (p = 0.005). Disease severity was not associated with survival times, but survival was related to younger age and male sex. In addition, ulcer diagnosis was a strong predictor of early termination. After adjustment for severity, survival times for RA and non-RA patients were the same. CONCLUSION: COX-2-specific inhibitors have a longer duration of use than NS NSAID. Among the COX-2-specific inhibitors, celecoxib has a longer survival time than rofecoxib. In addition, COX-2-specific inhibitors also have longer survival times than noted in the literature of NS NSAID in RA community practice. Duration of use can be an indicator of treatment effectiveness and/or drug acceptability, and provides additional interpretation beyond the results of clinical trials. PMID- 14760811 TI - Aspects of diurnal rhythmicity in pain, stiffness, and fatigue in patients with fibromyalgia. AB - OBJECTIVE: To determine diurnal rhythm characteristics of pain, stiffness, and fatigue in self-ratings performed by patients with fibromyalgia (FM). METHODS: Twenty-one women with FM made self-measurements of pain, stiffness, and fatigue on 100 mm horizontal visual analog scales at 6 prespecified timepoints at home for 10 consecutive days. Linear and multiple regressions were performed on the original data and the 24-hour means vs FM classifiers (age, disease duration, tender points, dolorimetry score, Fibromyalgia Impact Questionnaire score), respectively. Data were analyzed for 24-hour and 7-day time-effects by ANOVA and for diurnal and weekly rhythms by the cosinor technique. RESULTS: Individual ratings for pain, stiffness, and fatigue correlated highly with each other throughout the day and over the days of the week. Of the FM classifiers, dolorimetry score was found to be inversely related to the pain, stiffness, and fatigue scores. For the group of subjects with a low dolorimetry score (< 2.25 kg), a significant diurnal rhythm was found in each self-rated variable, with greater pain, stiffness, and fatigue observed in the morning and least in the late afternoon. No rhythm in pain or stiffness was observed in those subjects with a higher threshold for pain (dolorimetry score > 2.25 kg), while fatigue showed the same significant diurnal pattern as in the first group. For the group as a whole, the possible presence of a weekly variation was found with ratings for pain, stiffness, and fatigue higher on Sunday and Monday and lower on Friday. CONCLUSION: Ratings of pain, stiffness, and fatigue in FM are significantly correlated, and show diurnal and possibly weekly rhythmicity, especially when pain threshold is low (dolorimetry score < 2.25 kg), and are thus predictive of each other over these time spans. This has important implications for scheduling activities of daily living, for measurement in clinical trials, and possibly for timing the administration of medications. PMID- 14760810 TI - Functional imaging of pain in patients with primary fibromyalgia. AB - OBJECTIVE: To examine the function of the nociceptive system in patients with fibromyalgia (FM) using functional magnetic resonance imaging (fMRI). METHODS: Two groups of women, 9 with FM and 9 pain-free, volunteered to participate. In Experiment 1, we assessed psychophysical responses to painful stimuli and prepared participants for fMRI testing. For Experiment 2, subjects underwent fMRI scanning while receiving painful and nonpainful heat stimuli. Conventional and functional MR images were acquired using a 1.5 T MR scanner. Scanning occurred over 5 conditions. Condition 1 served as a practice session (no stimuli). Conditions 2 and 5 consisted of nonpainful warm stimuli. Conditions 3 and 4 consisted of an absolute thermal pain stimulus (47 degrees C) and a perceptually equivalent pain stimulus delivered in counterbalanced order. RESULTS: Experiment 1 indicated that subjects with FM were significantly more sensitive to experimental heat pain than controls (p < 0.001). In Experiment 2, fMRI data indicated that the FM group exhibited greater activity than controls over multiple brain regions in response to both nonpainful and painful stimuli (p < 0.01). Specifically, in response to nonpainful warm stimuli, FM subjects had significantly greater activity than controls in prefrontal, supplemental motor, insular, and anterior cingulate cortices (p < 0.01). In response to painful stimuli, FM subjects had greater activity in the contralateral insular cortex (p < 0.01). Data from the practice session indicated brain activity in pain-relevant areas for the FM group but not for controls. CONCLUSION: Our results provide further evidence for a physiological explanation for FM pain. PMID- 14760812 TI - International League of Associations for Rheumatology classification of juvenile idiopathic arthritis: second revision, Edmonton, 2001. PMID- 14760813 TI - Delayed diagnosis of Takayasu's arteritis: total abdominal aorta occlusion treated with axillo-bifemoral bypass. AB - Takayasu's arteritis is an inflammatory vasculitis that primarily affects the aorta and its major brances. Delayed diagnosis is common, and is largely attributed to the variable and nonspecific presentation of the initial symptoms. Involvement of the obdominal aorta causes severe claudication of the lower extremities, and bypass surgery is required in some cases. We describe a case in which diagnosis was delayed. Total occlusion of the obdominal aorta was successfully treated with axillobifemoral bypass. PMID- 14760814 TI - Fatal occlusive vessel disease in a patient with systemic juvenile idiopathic arthritis. PMID- 14760815 TI - No significant sex differences in temporal arteritis. PMID- 14760818 TI - Disease modifying antirheumatic drugs and pregnancy. PMID- 14760820 TI - "Soft" neurological signs in systemic lupus erythematosus. PMID- 14760822 TI - Validity of the scleroderma functional assessment questionnaire. PMID- 14760823 TI - Lack of association between ICAM-1 gene polymorphisms and biopsy-proven erythema nodosum. PMID- 14760824 TI - [Model core curriculum for dental education]. PMID- 14760825 TI - [Pharmacological education--current status and objectives in nursing education]. PMID- 14760826 TI - [Pelvic bone crest harvesting]. PMID- 14760827 TI - Recent publications in hematological oncology. PMID- 14760829 TI - Recent references. PMID- 14760830 TI - Current readings in nuclear medicine. PMID- 14760828 TI - Where are all the iron men and women? PMID- 14760831 TI - How do I choose a supplemental oxygen delivery device? PMID- 14760833 TI - Regarding"predictive factors and clinical consequences of proximal aortic neck dilatation in 230 patients undergoing abdominal aorta aneurysm repair with self expandable stent-grafts". PMID- 14760834 TI - Regarding"magnetic resonance angiography minimizes need for arteriography after inadequate carotid duplex ultrasound scanning". PMID- 14760832 TI - Regarding "ischemic intestinal involvement in a patient with Buerger disease: case report and literature review". PMID- 14760835 TI - Regarding"factors that predict prolonged length of stay after aortic surgery". PMID- 14760840 TI - [Hernia treatment]. PMID- 14760836 TI - Blasting off into e-business. PMID- 14760837 TI - The Dr. Jack E. Wells Memorial Dedication to Dentistry Award goes to Dr. Robert H. Montgomery. PMID- 14760838 TI - Re: "Surgical education: in need of a shift in paradigm". PMID- 14760839 TI - Re: "Inhibition of medullary thyroid carcinoma (MTC) cell proliferation and RET phosphorylation by tyrosine kinase inhibitors". PMID- 14760841 TI - [Surgical infection of soft tissues]. PMID- 14760842 TI - [A review of the study on Schefflera plants: chemical constituents and physiological activities]. PMID- 14760843 TI - [Advances in the study of amygdalin]. PMID- 14760845 TI - Preparation and characteristics of sol-gel-coated calix[4]arene fiber for solid phase microextraction. AB - 5,11,17,23-Tetra-tert-butyl-25,27-diethoxy-26,28-dihydroxycalix[4]arene/hydroxy terminated silicone oil coated fiber was first prepared and applied for solid phase microextraction (SPME) with sol-gel technology. The possible sol-gel mechanism was discussed and confirmed by IR spectra. It showed wonderful selectivity and sensitivity to polar (aromatic amines), nonpolar (benzene derivatives, polycyclic aromatic hydrocarbons) and high boiling point compounds (phthalates) and the extraction equilibria were reached quite fast. The coating has high thermal stability (380 degrees C) and solvent stability (organic and inorganic), thus its lifetime is longer than conventional fibers. In addition, it has surprising fiber-to-fiber and batch-to-batch reproducibility. The detection limits were quite low and the linear ranges were pretty broad for all analytes. PMID- 14760844 TI - Overview of the status and applications of capillary electrophoresis to the analysis of small molecules. AB - The status of capillary electrophoresis (CE) in the analysis of small molecules is reviewed and summarised with the illustrative use of recent literature references. Examples are cited in this review which demonstrate that CE is now a recognised and established technique in many industries, law courts and government regulatory agencies. Each of the principal areas of CE application in small molecule analysis are covered in sections which highlight the recent developments and possibilities within that area. Application areas include the analysis of pharmaceuticals, agrochemicals, chiral separations, and forensics is covered. This is an update to a previous review article [J. Chromatogr. A 856 (1999) 443] and covers papers published between 1999 and 2002. Technical developments and improvements, such as the advent of capillary array instrumentation for increased sample throughput, and improved detection options are described. Overall it is concluded that CE has become a recognised and established technique in many areas and is still within a period of development of both instrumentation and application which will continue to expand usage. PMID- 14760846 TI - Liquid chromatographic determination of trimethylamine in water. AB - A method for the selective determination of trimethylamine (TMA) in aqueous matrices by liquid chromatography is reported. The proposed procedure is based on the derivatization of the analyte with 9-fluorenylmethyl chloroformate (FMOC) in a precolumn (Hypersil C18, 30 microm, 20 mm x 2.1 mm i.d.) connected on-line to the analytical column (LiChrosphere 100 RP18, 5 microm, 125 mm x 4 mm i.d.). Gradient elution was performed with a mixture of acetonitrile-water-0.05 M borate buffer (pH 9.0). The method has been applied to the direct determination of TMA in water within the 0.25-10.0 microg/ml concentration interval, and can also be adapted to the determination of TMA over the range 0.05-1.0 microg/ml by incorporating a preconcentration stage with C18 solid-phase extraction (SPE) cartridges. Good linearity, reproducibility and accuracy was achieved within the tested concentration intervals. The limits of detection at 262 nm were 50 and 5 ng/ml for the direct method and for the method involving preconcentration, respectively. The proposed conditions allowed the selective determination of TMA in the presence of other primary and secondary short-chain aliphatic amines. The utility of the described procedure has been tested by determining TMA in different water samples. PMID- 14760848 TI - Thermodynamic studies of pressure-induced retention of peptides in reversed-phase liquid chromatography. AB - The pressure-induced retention of peptides on reversed-phase HPLC was studied by systematically changing organic solvent composition and temperature at both low (19 bar) and high (318 bar) pressures using a homologous series of hydrophobic poly-L-phenylalanine (n = 2-7) as the model compound. Based on van' t Hoff plots under different organic solvent compositions and pressures, the enthalpy change for the solute (deltaH) was determined. Moreover, both the enthalpy and entropy change for each phenylalanine residue (deltadeltaH and deltadeltaS), which corresponds to solute retention on a microenvironment along the depth of C18 chain, were also calculated by direct subtractions. Results indicate that under acetonitrile (ACN) compositions above 35%, the pressure caused deltadeltaS value to change from a negative to a positive value and both deltaH and deltadeltaH to change from a negative to a less negative value, all leading to a thermodynamic state closer to those under 35% acetonitrile composition. This implies that the pressure-induced retention observed in this study was an entropy-favored but enthalpy-unfavored process and was explained by pressure-induced desorption of solvent molecules that were associated with the stationary phase or with the peptide solute. Under 35% acetonitrile composition, however, it was found that neither deltadeltaH nor deltadeltaS value was significantly changed by the pressure. Whereas, both deltaH value and the intercept of van't Hoff plots under 35% acetonitrile composition were increased by pressure. This indicates that under low organic solvent composition, 35%, most of the acetonitrile molecules adsorbed on the surface of the stationary phase and only little solvent molecules were dissolved in the bulk stationary phase where the phenylalanine residues were partitioned. This study has provided new thermodynamic insights to the pressure induced retention for peptides and proteins. PMID- 14760847 TI - Single drop microextraction or solid phase microextraction-gas chromatography mass spectrometry for the determination of iodine in pharmaceuticals, iodized salt, milk powder and vegetables involving conversion into 4-iodo-N,N dimethylaniline. AB - A rapid sequence of oxidation and iodination using 2-iodosobenzoate as an oxidizing agent and N,N-dimethylaniline as an iodine scavenger at pH 6.4, when 4 iodo-N,N-dimethylaniline is formed, has been used for the determination of iodide by GC-MS. Solid phase microextraction (SPME) and single drop microextraction (SDME) have been used for the extraction of the iodo-derivative and their relative efficiencies compared. Pharmaceutical samples were subjected to solid phase extraction (SPE) for cleanup and the eluate analyzed for iodide. Iodate in salt samples was reduced to iodide with ascorbic acid. Milk powder and dried vegetables were wet combusted with peroxydisulfate to liberate covalently bound iodine as iodate which was reduced before derivatization. A rectilinear calibration graph was obtained for 0.1 microg-10 mg l(-1) iodide by both extraction methods, the correlation coefficient and limit of detection (LOD) were 0.9995 and 25 ng l(-1) iodide by SPME method, and 0.9998 and 10 ng l(-1) iodide by SDME method, respectively. SDME appeared to be more efficient technique than SPME for the present system. From the pooled data, the average recovery of spiked iodide to real samples was 100.7% (range 96.5-107.0%) with an average R.S.D. of 3.1% (range 2.6-4.5%). PMID- 14760849 TI - Chemical characterisation of different separation media based on agarose by static time-of-flight secondary ion mass spectrometry. AB - In this paper, the novel application of time-of-flight secondary ion mass spectrometry (TOF-SIMS) for qualitative and semi-quantitative investigation of the surface chemistry of separation media based on beaded agarose is reported. Five different media were studied: DEAE Sepharose Fast Flow, Q Sepharose Fast Flow, SP Sepharose Fast Flow, Phenyl Sepharose Fast Flow at ligand densities between 7 and 33% (w/w) and the base matrix Sepharose 6 Fast Flow. The obtained TOF-SIMS spectra reveal significant chemical information regarding the ligands (DEAE, Q, SP and Phenyl) which are covalently attached to the agarose-based matrix Sepharose 6 Fast Flow. For the anion-exchange media (DEAE and Q Sepharose Fast Flow), the positive TOF-SIMS spectra yielded several strong characteristic fragment peaks from the amine ligands. Structural information was obtained, e.g. from the peak at m/z 173.20, originating from the ion structure [(C2H5)2NCH2CH2NH(C2H5)2l+, which shows that the ligand in DEAE Sepharose Fast Flow is composed of both tertiary and quaternary amines. The positive spectrum of Phenyl Sepharose Fast Flow contained major fragments both from the base matrix and the ligand. The cation-exchanger (SP Sepharose Fast Flow) gave rise to a positive spectrum resembling that of the base matrix (Sepharose 6 Fast Flow) but with a different intensity pattern of the matrix fragments. In addition, peaks with low intensity at m/z 109.94, 125.94 and 139.95 corresponding to Na2SO2+, Na2SO3+ and Na2SO3CH2+, respectively, were observed. The positive TOF-SIMS spectrum of Sepharose 6 Fast Flow contains a large number of fragments in the mass range up to m/z 200 identified as CxHyOz and CxHy structures. The results clearly show that positive TOF-SIMS spectra of different media based on Sepharose 6 Fast Flow are strongly influenced by the ligand coupled to the matrix. The negative TOF-SIMS spectra contained several ligand-specific, characteristic peaks for the cation-exchanger, having sulphonate as the ion-exchange group. Negative fragments such as S-, SO-, SO2-, SO3-, C2H3SO3-, C3H5SO3- and OC3H5SO3- were observed. Phenyl Sepharose Fast Flow, which has an uncharged group (Phenyl) coupled to the agarose matrix yielded one ligand-related peak corresponding to the C6H5O- fragment. DEAE and Q ligands could only be identified by the appearance of the fragments CN- and CNO- in the negative spectrum. However, a strong peak corresponding to the counter ion (Cl-) was observed. TOF-SIMS analysis can also be used for the investigation of residues from the coupling procedure that bonds the ligands to the matrix. One example is the observation of bromine peaks in the negative spectrum of Q Sepharose Fast Flow. Furthermore, it has also been shown that different ligand concentrations of Phenyl Sepharose Fast Flow can easily be detected by TOF-SIMS analysis. Information regarding the difference between the ligand density on the surface of the beads and in the bulk can also be obtained. However, spectra registered on the outermost surface and on the pore surface (crushed beads) of DEAE Sepharose Fast Flow clearly show that the agarose and the DEAE groups are homogeneously distributed in the beads. PMID- 14760850 TI - Quantification of ochratoxin A in foods by a stable isotope dilution assay using high-performance liquid chromatography-tandem mass spectrometry. AB - A stable isotope dilution assay (SIDA) was developed for quantification of the mycotoxin ochratoxin A (OTA) by using [2H5]-OTA as internal standard. The synthesis of labelled OTA was accomplished by acid hydrolysis of unlabelled OTA and subsequent coupling one of the products, ochratoxin alpha, to [2H5]-L phenylalanine. The mycotoxin was quantified in foods by LC-tandem MS after extraction with buffers containing [2H5]-OTA and clean-up by immuno affinity chromatography or by solid phase extraction on silica. The method showed a sufficient sensitivity with a low detection and quantification limit of 0.5 and 1.4 microg/kg, respectively, and good precision in inter-assay studies showing a CV (n = 3) of 3.6%. The analysis of certified reference materials resulted in a low bias of 2.1% from the certified values and revealed excellent accuracy of the new method. To prove the suitability of SIDA. OTA was quantified in a number of food samples and resulted mainly in not detectable OTA contents. However, three samples of raisins exceeded the legal limit of 10 microg/kg and highlighted the need for further controlling the contamination with the mycotoxin. PMID- 14760851 TI - Evaluation of different liquid chromatography-electrospray mass spectrometry systems for the analysis of heterocyclic amines. AB - Three liquid chromatography-electrospray ionisation (LC-ESI) MS systems are evaluated for the analysis of heterocyclic amines (HAs). The electrospray sources and analysers (ion trap, single quadrupole and triple quadrupole) have been compared in terms of performance and quality parameters. In all cases, a C8 reversed-phase column and (acetic acid-ammonium acetate 30 mM pH 4.5) acetonitrile (ACN) as mobile phase were used. Ionisation source parameters, post column addition and working conditions for each acquisition mode (full scan, product ion scan, selected ion monitoring, and multiple reaction monitoring) were optimised for each instrument. The MS-MS spectra obtained with the ion trap and the triple quadrupole systems were very similar in both fragment ions and relative abundances, except for carbolines that showed adduct formation in the ion trap. Quality parameters were established and good precision (relative standard deviations (R.S.D.) < 12%) and very low limits of detection were obtained, mainly when using the triple quadrupole (< 9 pg injected). The content of HAs in a lyophilised beef extract was determined using the three instruments in order to compare their applicability for routine HAs analysis. PMID- 14760852 TI - Multidimensional analysis of denatured milk proteins by hydrophobic interaction chromatography coupled to a dynamic surface tension detector. AB - Multidimensional analysis of denatured milk proteins is reported using high performance liquid chromatography (HPLC) combined with dynamic surface tension detection (DSTD). A hydrophobic interaction chromatography (HIC) column (a TSK Gel Phenyl-5PW column, TosoBiosep), in the presence of 3.0 M guanidine hydrochloride (GdmHCl) as denaturing agent is employed as the mobile phase. Dynamic surface tension is measured through the differential pressure across the liquid-air interface of repeatedly growing and detaching drops. Continuous surface tension measurement throughout the entire drop growth (50 ms to 4 s) is achieved, for each eluting drop of 4 s length, providing insight into both the kinetic and thermodynamic behavior of molecular orientation processes at the liquid-air interface. An automated calibration procedure and data analysis method is applied with the DSTD system, which allows two unique solvents to be used, the HIC mobile phase for the sample and a second solvent (water for example) for the standard, permitting real-time dynamic surface tension data to be obtained. Three dimensional data is obtained, with surface tension as a function of drop time first converted to surface pressure, which is plotted as a function of the chromatographic elution time axis. Experiments were initially performed using flow injection analysis (FIA) with the DSTD system for investigating commercial single standard milk proteins (alpha-lactalbumin, beta-lactoglobulin, alpha-, beta-, kappa-casein and a casein mixture) denatured by GdmHCl. These FIA-DSTD experiments allowed the separation and detection conditions to be optimized for the HIC-DSTD experiments. Thus, the HIC-DSTD system has been optimized and successfully applied to the selective analysis of surface-active casein fractions (alpha s1- and beta-casein) in a commercial casein mixture, raw milk samples (cow's, ewe's and goat's milk) and other diary products (yogurt, stracchino, mozzarella, parmesan cheese and chocolate cream). The different samples were readily distinguished based upon the selectivity provided by the HIC-DSTD method. The selectivity advantage of using DSTD relative to absorbance detection is also demonstrated. PMID- 14760853 TI - A new multi-residue method for analysis of pesticide residues in fruit and vegetables using liquid chromatography with tandem mass spectrometric detection. AB - A new multi-residue method for determination of pesticide residues in a wide variety of fruit and vegetables, using the National Food Administration (NFA) ethyl acetate extraction and determination by means of LC-MS/MS, is presented. The method includes pesticides normally detected by LC-UV or LC-fluorescence such as benzimidazoles, carbamates, N-methylcarbamates and organophosphorus compounds with an oxidisable sulphide group as well. After extraction with ethyl acetate, the extract is concentrated and an aliquot of the extract is evaporated to dryness and redissolved in methanol before injection on LC-MS/MS. The method has been validated for 57 different pesticides and metabolites. Representative species from different commodity groups were chosen as matrices in order to study the influence from different matrices on recoveries. The fortification levels studied were 0.01-0.5 mg kg(-1). Matrix effects were tested for all matrices by means of standard addition to blank extracts. The matrix effect, expressed as signal in solvent compared to signal in matrix, was in general found to be small. The obtained recoveries are, with a few exceptions, in the range 70-100%. The proposed method is quick and straightforward and no additional clean-up steps are needed. The method can be used for the analysis of all 57 pesticides in one single determination step at 0.01 mg kg(-1). PMID- 14760854 TI - Ion chromatographic determination of sulfide and cyanide in real matrices by using pulsed amperometric detection on a silver electrode. AB - The determination of free sulfide and cyanide by pulsed amperometric detection (PAD) at a silver-working electrode was improved through a deep de-oxygenation (at least 10 min) of both standard and real solutions containing the two analytes and adopting a two-potential waveform able to eliminate Ag working electrode fouling. The waveform stepped around the oxidation of Ag in the presence of 0.1 0.4 M hydroxyl ion, from -0.1 to 0.1 V versus saturated calomel electrode (SCE). The eluent composition (0.4 M NaOH plus 7.5 mM oxalate solution) allowed a very good column efficiency and selectivity. The presence of a polysulfide species was hypothesized in sulfide solutions that had not been de-oxygenated and aged. The polysulfide eluted just before sulfide and was confirmed by a chemical test with SO3(2-) producing the elimination of the polysulfide peak. Detection limits, according to the Hubaux-Vos method, were 1.0 and 2.0 microg/l for S2- and CN , respectively. We demonstrated good performance of the optimized method by repeatedly injecting standard solutions and by analyzing different real matrices. The method exhibited very good accuracy and repeatability (10 microg/l and a 500 microl injection loop, had a repeatability better than 3% for sulfide and 100 microg/l had a repeatability better than 1% for cyanide). The two-potential waveform ensured long-term stability of the electrode surface that required no manual polishing procedure for at least 1 month (20 analysis per day). PMID- 14760855 TI - Evaluation of a structure-driven retention model for temperature-programmed gas chromatography. AB - The solvation parameter model is suitable for describing the retention properties of compounds of varied structure in temperature-programmed gas chromatography. An empirical second-order model provides a good account of the change in system constants as a function of program rate. These relationships codify the reduction in retention time at higher program rates and changes in elution order (selectivity) with program rate. The prediction of retention times from structure, while quite good, is probably adversely affected by descriptor quality and the possibility of a mixed retention mechanism on polar stationary phases. Plots of experimental against predicted temperature-programmed retention times for varied compounds are linear but generally contain a small bias from an ideal model (slope of one and an intercept of zero). The average absolute deviation in temperature-programmed retention times on three columns (DB-210, DB-1701 and EC Wax) varied from 0.15 to 0.89 min with the best results obtained at higher program rates on the columns of lower polarity. PMID- 14760856 TI - Stability test and improvement of hydrogen analyzer with trace reduction detector. AB - We previously developed an analyzer able to detect hydrogen concentrations of less than 50 cm3/1000 m3. The analyzer uses a carrier gas purifier and a low temperature separation column to remove impurities preventing measurement of low concentrations from the carrier and sample gases. It uses a trace reduction detector with a mercuric oxide bed to detect the concentration of hydrogen based on the reduction reaction of mercuric oxide with hydrogen. We have now evaluated the performance of the analyzer by carrying out a series of tests that measured the spectrum peak and the retention time. We used three sample gases with hydrogen concentrations of 5, 20, and 50 cm3/1000 m3 in nitrogen dilution gas. The measured peak was stable (it was within a relative standard deviation of less than 10%), and there was a linear relationship between the peak and hydrogen concentration. However, the retention time gradually shortened as the measurements were repeated. The shortening was reduced by warming the low temperature separation column used in the analyzer; it was not observed when we used a hydrogen sample gas diluted by helium instead of nitrogen. Using nitrogen as a dilution gas apparently shortens the retention time. We thus added an MS-5A separation column and a thermal conductivity detector. The nitrogen and hydrogen in the sample/carrier gas are separated, and the nitrogen is efficiently removed by switching the pass line to a release line after the hydrogen has been sent to the low temperature separation column. An analyzer using this "after-cut method" was able to stably measure infinitesimal hydrogen concentrations and was not affected by nitrogen in the sample gas. PMID- 14760857 TI - Comparative study for separation of aquatic humic substances by capillary zone electrophoresis using uncoated, polymer coated and gel-filled capillaries. AB - Several comparative capillary zone electrophoresis (CZE) experiments were carried out by means of uncoated, polyvinyl alcohol (PVA) and polyacrylamide (PAA) coated silica open tubular capillaries and gel-filled capillaries (linear non-cross linked polyacrylamide, PAGE, by a pre-coated PAA capillary) using different kinds of background electrolytes (BGEs) and organic modifiers for characterization of aquatic dissolved humic matter (DHM). Organic compounds, such as acetic acid, acetate buffer, methanol, ethylene glycol, acetonitrile, dimethylsulphoxide, 5 M urea and sodium dodecyl sulphate (SDS) were tested as sample modifiers to improve the separative power. The fractionation mode by a PVA coated open tubular capillary using 40 mM phosphate buffer at pH 6.8 and 5 M urea-water as the sample modifier turned out to be fairly practical as well as its PAA homologue. Linear non-cross-linked PAGE with 10% gel concentration and 5 M urea-water as the sample modifier using 40 mM phosphate buffer at pH 6.8 produced the most reliable results as to the adaptation of physical gels, especially if the interactions of humic solutes with the gel matrix are not critical. The addition of SDS in the linear PAGE gel increased the interaction of humic solutes with the gel matrix but also improved the separative power and strengthened the chaotropic effect of the urea modifier. PMID- 14760858 TI - In-channel dual-electrode amperometric detection in electrophoretic chips with a palladium film decoupler. AB - An electrophoretic microchip integrated with a Pd-film decoupler and a series dual electrode was proven practical (200-800 V/cm) for routine amperometric detection. In fluidic systems, amperometric enhancement of parallel-opposed dual electrode detection is due to redox cycling of analytes between the electrodes. We, however, found that the oxidation current of catecholamines was enhanced significantly (1.9-3.8 folds) by switching from the single electrode mode to dual series mode. This novel finding was unexpected because the unidirectional flow characteristic of the microfluidic system should eliminate the possibility for analytes physically migrating back and forth between the upstream and downstream electrodes. We attribute the enhancement to turbulence generated by impinging of the flow onto the edge of the downstream electrode. The linear range, sensitivity, limit of detection (S/N = 3) and number of theoretical plates for DA and CA are, respectively, 0.5-50 microM, 47 pA/microM, 0.25 microM, 7000 m(-1) and 1.0-100 microM, 28 pA/microM, 0.49 microM, 15,000 m(-1). PMID- 14760859 TI - On-column complexation capillary electrophoretic separation of Fe2+ and Fe3+ using 2,6-pyridinedicarboxylic acid coupled with large-volume sample stacking. AB - On-column complexation of Fe2+ and Fe3+ with 2,6-pyridinedicarboxylic acid (2,6 PDCA) formed anionic complexes, which were then separated by capillary zone electrophoresis with direct UV detection at 214 nm. To achieve reasonable separation selectivity and on-column complexation, the conditions such as pH, the concentration of 2,6-PCDA and the EOF modifiers in the electrolyte were examined. The electrolyte contained 5.0 mM 2,6-PDCA, 0.25 mM tetradecyltrimethlammonium bromide (TTAB) and 5% (v/v) acetonitrile at pH 4.0 was optimised for on-column complexation and the separation of Fe[PCDA]2(2-) and Fe[PCDA]2(-). To enhance the detection sensitivity, large-volume sample stacking (LVSS) was used for the on line preconcentration of Fe[PCDA]2(2-) and Fe[PCDA]2(-). Under the optimised conditions, satisfactory working ranges (0.5-50 microM), lower detection limits (less than 0.1 microM) and good repeatability of the peak areas (R.S.D.: 5.2 7.8%, n = 5) was achieved using LVSS (300 s). With LVSS, the detection sensitivity was enhanced more than 50-fold compared to conventional hydrodynamic injection. The proposed method was used successfully for the determination of Fe2+ and Fe3+ in water samples. PMID- 14760860 TI - Preparative high-speed counter-current chromatography for purification of shikonin from the Chinese medicinal plant Lithospermum erythrorhizon. AB - The bioactive compound shikonin was successfully isolated and purified from the crude extract of the traditional Chinese medicinal plant Lithospermum erythrorhizon Sieb. et Zucc. by preparative high-speed counter-current chromatography (HSCCC). The preparative HSCCC was performed using a two-phase solvent system composed of n-hexane-ethylacetate-ethanol-water (16:14:14:5 (v/v)). A total amount of 19.6 mg of shikonin at 98.9% purity was obtained from 52 mg of the crude extract (containing 38.9% shikonin) with 96.9% recovery. The preparative isolation and purification of shikonin by HSCCC was completed in 200 min in a one-step separation. PMID- 14760861 TI - Structure-activity relationship for alkylating dipeptide nitrogen mustard derivatives. AB - The strategy of using small peptides for effective targeting of tumor cells in chemotherapy has proven beneficial. Recently we showed that J1 (L-melphalanyl-p-L fluorophenylalanine ethyl ester), an alkylating nitrogen mustard-containing dipeptide, exhibited strong cytotoxic activity in fresh human tumor samples in addition to rapid and pronounced inhibition of macromolecular syntheses and cellular respiration in the human tumor lymphoma cell line U-937 GTB. In this study, an additional series of 17 nitrogen mustard-containing dipeptides has been synthesized and analyzed for cytotoxic activity in a panel of 10 human tumor cell lines. The results were compared to the single amino acid mustard derivative melphalan and its ethyl and isopropyl esters. Also P2 (L-prolyl-m-L-sarcolysyl-p L-fluorophenylalanine ethyl ester), a tripeptide that previously has shown impressive effects in human tumor cells, was used as reference. The tested compounds displayed various activities in the different cell lines but also showed a high correlation, indicating a similar mechanism of action. Factors like amino acid composition, amino acid sequence, modifications of the C- and N termini, and to a minor extent the lipophilicity of the dipeptide derivatives appear to influence the in vitro activity. The results indicate that the activity of these compounds not only relies on their chemical reactivity, but also on active biological interactions such as transport across membranes and/or enzymatic liberation of reactive molecular entities. PMID- 14760862 TI - Shifting gene expression profiles during ex vivo culture of renal tumor cells: implications for cancer immunotherapy. AB - The use of cultured tumor cells rather than original tumor tissue for the preparation of therapeutic cancer vaccines represents an obvious solution to the problem of availability of adequate quantities of autologous tumor. In this study we investigated possible changes in gene expression accompanying the transition of renal cell carcinoma cells from the original tissue to cell populations in culture. In our study we employed cDNA microarray technology to compare the gene expression pattern of ex vivo cultured renal carcinoma cells to that of the original solid tumor tissue from which the cells were derived. Using this approach we detected changes in the expression of many genes mostly related to the cell lines' physiological properties. Some of the products of those genes showing differential expression between tumor-derived cell line and original tumor are known human autoantigens or tumor-associated antigens. Furthermore, analysis of overexpressed genes revealed the presence of several transcripts with restricted normal tissue distribution, representing self-antigens with potential to elicit autoimmunity. Our results suggest that adapting tumor tissue to culture can result in changes in the level of transcripts specific for known antigens and that more information regarding the composition of tumor cells and their byproducts used in vaccine trials is needed before the efficacy and safety of such procedures can truly be determined. PMID- 14760863 TI - In vitro antitumor structure-activity relationships of threo/trans/threo mono tetrahydrofuranic acetogenins: correlations with their inhibition of mitochondrial complex I. AB - In this study we evaluated a mono-tetrahydrofuranic subgroup of natural acetogenins that had shown in previous enzyme inhibition studies different potency trends compared with the bis-tetrahydrofuranic acetogenin subgroup. The compounds were tested against colon, breast, lung, liver, and ovarian tumor cell lines. A drug-resistant ovarian cell line was also included in the panel. In general the compounds were more potent than doxorubicin. The goal was to determine how well the mitochondrial complex I inhibition correlates with the in vitro antitumor potency of these natural mono-tetrahydrofuranic acetogenins and of some derivatives. The results indicate that both the reduction of the terminal gamma-lactone after its translactonization and the introduction of an hydroxylimine group in the alkyl chain, near the mono-tetrahydrofuranic moiety, increased the antitumor activity, even against the doxorubicin-resistant cell line. PMID- 14760864 TI - Pleural mesothelial cell (PMC) defense mechanisms against malignancy. AB - Tumors such as ovarian, lung, and breast have been found to have a predilection for the pleura. Pleural mesothelial cells (PMCs) play an active role in pleural inflammation via release of cytokines. However, mechanisms whereby PMCs defend themselves against invading malignant cells are unknown. In the present study, we hypothesized that PMCs release the antiangiogenic factor endostatin and inhibit malignant cell invasion. We evaluated the endostatin levels in malignant (MAL) and congestive heart failure (CHF) pleural fluids (PF). Endostatin expression by PMC was also demonstrated by Western analysis and confocal microscopy. Our results demonstrate that CHF PF contained significantly higher levels of endostatin when compared with MAL PF. PMCs alone released a significantly greater amount of endostatin when compared with ovarian cancer cells (OCCs). When the PMC were cocultured with OCCs without contact, there was an increase in the endostatin production. However, when the PMCs were cocultured in direct contact with OCCs the endostatin levels significantly decreased. Endostatin production was upregulated in the presence of tumor cells but not when OCCs were adherent to underlying PMC monolayer. Immunofluorescent staining of PMCs for endostatin correlated with endostatin release. These findings suggest that PMCs play a key role in the antiangiogenesis process by producing endostatin in the pleural space, and thus preventing tumor spread and metastasis in the pleura. PMID- 14760865 TI - Expression of c-Met proto-oncogene in metastatic macrophage x melanoma fusion hybrids: implication of its possible role in MSH-induced motility. AB - It was shown previously that a majority of hybrids produced by in vitro fusion of normal macrophages with Cloudman S91 melanoma cells displayed macrophage-specific glycosylation, especially increased GnT-V activity, beta1,6 branch formation in glycoproteins, accompanied by enhanced metastatic potential in vivo and motility in vitro. These hybrids also express upregulated melanocortin-1 receptor (MC1-R) activity and exhibit increased motility after melanocyte-stimulating hormone (MSH) treatment. In this report, we show that MSH-mediated stimulation of motility is mediated through enhanced expression of c-Met proto-oncogene. In metastatic hybrids c-Met expression is induced by MSH, and addition of c-Met neutralizing antibody to cells inhibits MSH-induced motility but not the basal motility of the cells. Furthermore, abrogation of the chemoattractant gradient concentration by addition of hepatocyte growth factor (HGF) recombinant protein, a cognate ligand of c-Met receptor, reduces the MSH-induced effect on motility. A similar result was also obtained by the addition of blocking anti-alphaHGF antibody in the chemoattractant chamber. Again, the metastatic hybrids, but not the nonmetastatic hybrids or parental melanoma cells, showed significant motile response to rHGF chemoattractant, and that motility is further induced when cells were stimulated with MSH/isobutylmethyl xanthine (IBMX). Synergistic stimulation on motility was also observed with those hybrids treated with MSH/IBMX and when rHGF and fibronectin (FN), in combination, were used as chemoattractants. These indicate that MSH/IBMX-induced motility might involve c-Met pathways as well as extracellular matrix (ECM)/integrin pathways in a cooperative fashion. Ets-1, a transcription factor involved in the expression of c-Met, is also found to be induced in metastatic hybrids after exposure to MSH/IBMX. Implication of the result is discussed in light of the role of c-Met and its interacting proteins in the development of metastatic phenotypes and its therapeutic intervention. PMID- 14760866 TI - Autonomy, subject-relativity, and subjective and objective theories of well-being in bioethics. AB - Among the different approaches to questions of biomedical ethics, there is a view that stresses the importance of a patient's right to make her own decisions in evaluative questions concerning her own well-being. This approach, the autonomy based approach to biomedical ethics, has usually led to the adoption of a subjective theory of well-being on the basis of its commitment to the value of autonomy and to the view that well-being is always relative to a subject. In this article, it is argued that these two commitments need not lead to subjectivism concerning the nature of well-being. PMID- 14760867 TI - Rawls and research on cognitively impaired patients: a reply to Maio. AB - In his paper, "The Relevance of Rawls' Principle of Justice for Research on Cognitively Impaired Patients" (Theoretical Medicine and Bioethics 23 (2002): 45 53), Giovanni Maio has developed a thought-provoking argument for the permissibility of non-therapeutic research on cognitively impaired patients. Maio argues that his conclusion follows from the acceptance of John Rawls's principles of justice, specifically, Rawls's "liberty principle". Maio has misinterpreted Rawls's "liberty principle"--correctly interpreted it does not support non therapeutic research on cognitively impaired patients. Three other 'Rawlsian' arguments are suggested by Maio's discussion--two "self-respect" arguments and a "presumed consent" argument--but none of them are convincing. However, an alternative argument developed from Rawls's discussion of "justice in health care" in his most recent book, Justice as Fairness: A Restatement, may justify certain kinds of non-therapeutic research on some cognitively impaired patients in special circumstances. We should not expect anything more permissive from a liberal theory of justice. PMID- 14760870 TI - What can we learn by looking for the first code of professional ethics? AB - The first code of professional ethics must: (1) be a code of ethics; (2) apply to members of a profession; (3) apply to all members of that profession; and (4) apply only to members of that profession. The value of these criteria depends on how we define "code", "ethics", and "profession", terms the literature on professions has defined in many ways. This paper applies one set of definitions of "code", "ethics", and "profession" to a part of what we now know of the history of professions, thereby illustrating how the choice of definition can alter substantially both our answer to the question of which came first and (more importantly) our understanding of professional codes (and the professions that adopt them). Because most who write on codes of professional ethics seem to take for granted that physicians produced the first professional code, whether the Hippocratic Oath, Percival's Medical Ethics, the 1847 Code of Ethics of the American Medical Association (AMA), or some other document, I focus my discussion on these codes. PMID- 14760869 TI - Hermeneutics of medicine in the wake of Gadamer: the issue of phronesis. AB - The relevance of the Aristotelian concept of phronesis--practical wisdom--for medicine and medical ethics has been much debated during the last two decades. This paper attempts to show how Aristotle's practical philosophy was of central importance to Hans-Georg Gadamer and to the development of his philosophical hermeneutics, and how, accordingly, the concept of phronesis will be central to a Gadamerian hermeneutics of medicine. If medical practice is conceived of as an interpretative meeting between doctor and patient with the aim of restoring the health of the latter, then phronesis is the mark of the good physician, who through interpretation comes to know the best thing to do for this particular patient at this particular time. The potential fruitfulness of this hermeneutical appropriation of phronesis for the field of medical ethics is also discussed. The concept can be (and has been) used in critiques of the conceptualization of bioethics as the application of principle-based theory to clinical situations, since Aristotle's point is exactly that problems of praxis cannot be approached in this way. It can also point the way for alternative forms of medical ethics, such as virtue ethics or a phenomenological and hermeneutical ethics. The latter alternative would have to address the phenomena of health and the good life as issues for medical practice. It would also have to map out in detail the terrain of the medical meeting and the acts of interpretation through which phronesis is exercised. PMID- 14760871 TI - Clinical and laboratory diagnosis of acute renal failure. AB - Acute renal failure (ARF) is defined in general terms as an abrupt decrease in renal function sufficient enough to result in retention of nitrogenous waste and disrupt fluid and electrolyte homeostasis. There is no consensus regarding a quantifiable definition of ARF. Prompt evaluation of ARF is vital because ARF can be the end result of diverse processes which can often be reversed or attenuated through therapy directed at the underlying condition. Evaluation begins with careful review of the patient's history, previous medical records, physical examination, urinalysis, and available laboratory data. Routine urine chemical indices, calculation of the fractional excretion of sodium, and examination of the urine sediment are valuable in characterizing the cause of ARF. When this evaluation fails to yield a diagnosis, further testing may be required to evaluate intravascular volume status or diagnose a systemic disorder or glomerular cause of ARF. Response to therapeutic trials may provide a diagnosis. When a diagnosis cannot be made with reasonable certainty through this evaluation renal biopsy should be considered. PMID- 14760872 TI - Pathophysiology of ischaemic acute renal failure. AB - This chapter summarizes the pathophysiology of ischaemic acute renal failure from both the experimental and clinical points of view. Traditionally, the abrupt fall in glomerular filtration rate (GFR) is thought to be due to an interplay of haemodynamic and tubular abnormalities. The intrarenal haemodynamic alterations include renal vasoconstriction, leukocyte-endothelium interactions and loss of blood flow and GFR autoregulation. During recent years it has become evident that pronounced outer medulary ischaemia makes an important contribution. In severe and prolonged ischaemia, the tubular epithelial cells can undergo either sublethal or lethal cell damage. Cell death occurs by necrosis and apoptosis. The different mechanisms of post-ischaemic cell damage are discussed. The post ischaemic kidney also shows a dramatic capacity for recovery. During this recovery phase some of the damaged cells undergo de-differentiation--which is an important step in regeneration of the tubular epithelium. Recent evidence points to the possibility that infiltration of the kidney with bone-marrow-derived stem cells contributes to the repair process. The molecular mechanisms and the effect of growth factors are summarized. PMID- 14760873 TI - Acute toxic renal failure. AB - Acute renal failure (ARF) is a common problem in intensive care medicine. Even modest degrees of ARF not requiring dialysis treatment increase the risk of death approximately fivefold. Despite the widespread appreciation of the role of nephrotoxic drugs in their contribution to ARF, these drugs continue to have an ongoing aetiological role. Potentially nephrotoxic drugs include non-steroidal anti-inflammatory drugs, radiocontrast agents, antimicrobial and anaesthetic agents. Endogenous compounds such as myoglobin and haemoglobin may furthermore cause toxic nephropathy. Tubular injury initiated by toxins often results from a combination of acute renal vasoconstriction and direct cellular toxicity due to intracellular accumulation of the toxin, or, alternatively, may be mediated immunologically in case of interstitial nephritis. Patients with reduced renal functional reserve, cardiovascular co-morbidity, diabetes mellitus, and advanced age are at increased risk. Awareness of the range of toxins on the one hand and simple measures such as adequate pre-hydration of the patient and drug monitoring on the other hand may be sufficient to avoid drug-induced ARF or minimize its clinical severity in susceptible patients. PMID- 14760874 TI - Vasoactive drugs and the kidney. AB - Protection of renal function and prevention of acute renal failure (ARF) are important goals of resuscitation in critically ill patients. Beyond fluid resuscitation and avoidance of nephrotoxins, little is known about how such prevention can be achieved. Vasoactive drugs are often administered to improve either cardiac output or mean arterial pressure in the hope that renal blood flow will also be improved and, thereby, renal protection achieved. Some of these drugs (especially low-dose dopamine) have even been proposed to have a specific beneficial effect on renal blood flow. However, when all studies dealing with vasoactive drugs and their effects on the kidney are reviewed, it is clear that none have been demonstrated to achieve clinically important benefits in terms of renal protection. It is also clear that, with the exception of low-dose dopamine, there have been no randomized controlled trials of sufficient statistical power to detect differences in clinically meaningful outcomes. In the absence of such data, all that is available is based on limited physiological gains (changes in renal blood flow or urine output) with one or another drug in one or another subpopulation of patients. Furthermore, given our lack of understanding of the pathogenesis of ARF, it is unclear whether haemodynamic manipulation is an appropriate avenue to achieve renal protection. There is a great need for large randomized controlled trials to test the clinical, instead of physiological, effects of vasoactive drugs in critical illness. PMID- 14760875 TI - Should we use diuretics in acute renal failure? AB - Because oliguria is a bad prognostic sign in patients with acute renal failure (ARF), diuretics are often used to increase urine output in patients with or at risk of ARF. From a pathophysiological point of view there are several reasons to expect that loop diuretics also could have a beneficial effect on renal function. However, clinical trials on the prophylactic use of loop diuretics rather point to a deleterious effect on parameters of kidney function. In patients with established ARF loop diuretics have been shown to increase urine output, which may facilitate patient management. A beneficial effect on renal function has, however, not been demonstrated. On the other hand, such an effect cannot be excluded because the available trials lack statistical power. Possible explanations for the absence of a renoprotective effect are discussed. The evidence for a renoprotective effect of mannitol is restricted to the setting of renal transplantation. PMID- 14760876 TI - Prevention of perioperative acute renal failure: what works? AB - Perioperative acute renal failure (ARF) is associated with increased morbidity and mortality. Patients undergoing cardiac, vascular and major abdominal surgery and those with pre-operative renal insufficiency are at increased risk for developing post-operative ARF. The aetiologies of perioperative ARF are multi factorial. However, pre-renal azotaemia and ischaemic acute tubular necrosis (ATN) are the predominant causes. Preventive strategies involve identifying patients at risk, optimizing intravascular volume as well as renal function with perioperative haemodynamic monitoring, and avoiding nephrotoxins. Various pharmacological agents have been used to optimize renal perfusion and tubular function. Unfortunately, none has been shown to be effective in randomized placebo-controlled trials. In this chapter, we discuss the prophylactic use of fluids, vasoactive drugs, diuretics and other agents, as well as modification of surgical techniques to reduce the incidence of perioperative ARF. PMID- 14760878 TI - Perioperative management of patients with chronic kidney disease or ESRD. AB - The perioperative management of patients with chronic kidney disease (CKD) or dialysis-dependent end-stage renal disease (ESRD) is complicated by both the underlying renal dysfunction, with associated disturbances of fluid and electrolyte homeostasis and altered drug clearance, and the presence of associated co-morbid conditions, including diabetes mellitus, chronic hypertension and cardiovascular and cerebrovascular disease. The impact of CKD on fluid and electrolyte management, haematological and cardiovascular complications and drug management in the perioperative period are reviewed. Special issues related to the management of haemodialysis and peritoneal dialysis patients in the perioperative period are also reviewed. PMID- 14760877 TI - Acid-base balance in acute renal failure and renal replacement therapy. AB - The approach to acid-base balance based on the concept of strong ions, initially proposed by Stewart, is briefly overviewed. The anion gap and the strong anion gap are both discussed. Comments are made on the strong ion difference of fluids administered to patients and their impact on acid-base status will be commented. Renal failure patients have an altered acid-base balance; most commonly, a mixed type of metabolic acidosis (hyperchloraemic, and of a high anion gap) is observed. The consequences of renal metabolic acidosis are described. Finally, the impact of renal replacement therapy on acid-base balance is exposed; different modalities of renal replacement are considered in regard to their alkalinizing performance. PMID- 14760879 TI - Renal replacement therapy in acute renal failure. AB - Acute renal failure (ARF) is a common condition in hospitalized patients. The aetiology and physiological characteristics differ from those of chronic renal failure (CRF) and both conditions should be approached differently. At present, the approach to the management of ARF is very heterogeneous. Attempts have therefore been made to improve consensus and to standardize treatment in the Acute Dialysis Quality Initiative (ADQI). Technology for the treatment of ARF is expanding. Traditional intermittent haemodialysis (IHD) is still a major treatment modality but continuous renal replacement therapies (CRRT) and slow, low-efficiency daily dialysis (SLEDD) are commonly used alternatives. Each modality has advantages and disadvantages, but to date no evidence exists for the superiority of one over the other. On the other hand, the availability of multiple options allows us to provide tailor-made treatment: the best modality is chosen depending on local expertise and the individual clinical scenario. Practice guidelines based on the best available evidence and the author's opinion are suggested. PMID- 14760880 TI - Vascular access for dialysis in the intensive care unit. AB - Management of the vascular access (VA) for renal replacement therapy (RRT) in acute renal failure (ARF) patients is faced with a twofold problem: first, the creation of an angio-access that is adequate for RRT in the acute setting; second, the preservation of the patient's vascular network in order not to preclude further use of the vessel in the event of evolution to chronic renal failure. Central venous catheters are the preferred VA for RRT in the intensive care setting. Semi-rigid double-lumen polyurethane catheters may be considered for short-time use (up to 2-3 weeks). Soft silicone double-lumen or twin catheters, preferably with subcutaneous tunnelling, are highly desirable for prolonged RRT (over 3 weeks). The femoral route is the first option in the presence of associated risk factors (respiratory failure, pulmonary oedema, bleeding...). The internal jugular route should be considered for mid-term use in order to facilitate the patient's mobilization and to reduce the risk of infection. The subclavian route should be avoided because of the risk of stenosis and/or thrombosis of the outflow vein. Catheter insertion must be performed by a trained physician with ultrasound guidance using either skin mapping or continuous vein guidance. Catheter handling and care should comply with best practice guidelines and should be part of a continuous quality improvement programme in order to reduce catheter-related morbidity. Preservation of the upper limb vascular network of the patient consists of sparing the native vessels (artery and vein) of the patient and preserving the functionality of the permanent VA in chronic renal failure patient. This 'lifeline' of chronic renal failure patients may be maintained by preventing inflammation, infection and thrombosis of the superficial vessels of the arm and forearm of patient. PMID- 14760881 TI - Influence of renal replacement therapy on pharmacokinetics in critically ill patients. AB - Critical illness has a great impact on many pharmacokinetic parameters. An increased volume of distribution often results in drug underdosing, whereas organ impairment may lead to drug accumulation and overdosing. Renal replacement therapy (RRT) in critically ill patients with renal failure may significantly increase drug clearance, requiring drug-dosing adjustments. Drugs significantly eliminated by the kidney are likely to experience substantial removal during RRT, and a supplemental dose--corresponding to the amount of drug removed by RRT- should be administered. Mechanisms of drug removal during RRT are reviewed together with methods for measuring or estimating RRT drug clearances. Approaches for drug-dosing adjustments are suggested and, at the end, the pharmacological principles for antibiotic prescription in the critically ill are discussed. PMID- 14760882 TI - Effect of mechanical ventilation on the kidney. AB - Mechanical ventilation is a standard component of intensive care unit management of critically ill patients and is widely used for respiratory support. Recent animal and clinical studies have shown that positive pressure ventilation can worsen pre-existing lung injury and produce ventilator-induced lung injury, which has been linked with the development of systemic inflammation and multi-system organ dysfunction, including renal failure. Although the physiological consequences of mechanical ventilation on pulmonary and cardiovascular function have been extensively studied, its effects on renal function are not as well defined. Previous experimental studies and few clinical reports have shown a significant effect of mechanical ventilation on renal function. Interestingly, recent data are emerging which suggest that renal dysfunction also has a direct, adverse effect on pulmonary function. This chapter reviews the information in these areas and provides a framework for future investigation in this field. PMID- 14760883 TI - Nucleoside and nucleotide analogue reverse transcriptase inhibitors: a clinical review of antiretroviral resistance. AB - Although advances in highly active antiretroviral therapy (HAART) have made long term suppression of HIV an achievable goal of therapy, a substantial proportion of first-line regimens will eventually fail. Successful longterm treatment requires consideration of downstream treatment options at the time of initiating or changing regimens. An understanding of the patterns and interactions of resistance mutations, and the appropriate use of genotypic and phenotypic testing is an important component of successful drug sequencing. Resistance to multiple nucleoside reverse transcriptase inhibitors (NRTIs) may result from several genotypically distinct pathways, including the Q151M (151 complex), the 69 insertion complex, two distinct thymidine analogue mutational pathways and the K65R mutation. Knowledge of the clinical implications of these and other resistance pathways, as well as the antagonism or synergy between mutations, helps guide individualized treatment choices from initial therapy in the treatment-naive patient to salvage therapy in the highly treatment-experienced individual. The development of effective sequencing strategies will depend upon the continued understanding of drug resistance mutation patterns and their associations with specific HAART combinations. This review summarizes research advances that further the understanding of nucleoside and nucleotide analogue resistance mutations, and their interplay. PMID- 14760887 TI - The potentiating effect of ribavirin on interferon in the treatment of hepatitis C: lack of evidence for ribavirin-induced viral mutagenesis. AB - The recent finding that ribavirin has a mutagenic capacity in a poliovirus replicon model pushing the virus into error catastrophe, provides a possible explanation for the remarkable synergistic effect of ribavirin when combined with interferon in the treatment of chronic hepatitis C virus (HCV)-infected patients. However, ribavirin-induced hypermutation resulting in loss of vital genetic information and viral clearance, does not occur during treatment of HCV-infected patients, as can be inferred from the lack of viral inhibition when treating HCV infected patients with ribavirin alone. We therefore hypothesized that ribavirin induces mutations in the C-terminal part of the viral NS5A gene, a region found to be correlated with interferon sensitivity. Ribavirin-induced mutations resulting in the appearance of viral variants more sensitive to interferon would explain the synergistic effect of ribavirin when combined with interferon. To test this hypothesis we retrospectively analysed sequences of the C-terminal half of the NS5A gene before and during treatment in six HCV genotype 1-infected patients who had been treated with combination therapy after initial failure to respond permanently to interferon alone. Our results show that during the early treatment phase mutation rate is not enhanced during combination therapy and that, at least in the major variant, shifts in the NS5A domain resulting in the occurrence of viral variants, which are more interferon-sensitive, do not occur. PMID- 14760885 TI - Sustained response to interferon-ribavirin combination therapy predicted by a model of hepatitis C virus dynamics using both HCV RNA and alanine aminotransferase. AB - BACKGROUND AND AIMS: Standard models simulate the dynamics of hepatitis C virus (HCV) infection using HCV RNA kinetics and show a correlation between the clearance of infected hepatocytes and response to interferon. However, they are unable to predict whether the response will be maintained. The aim of our work was to identify criteria by which sustained responses may be predicted and defined. METHODS: In our model the clearance rate of infected cells is a function of their number and the baseline rate is computed by the alanine aminotransferase (ALT) kinetics during the first month of therapy. We simulated the variations of viral and infected cell loads in 31 consecutive patients treated with IFN-alpha2b 3-5 MU every other day, with or without ribavirin, for 6 or 12 months depending on HCV genotype. RESULTS: At baseline the computed (in 28 of 31 cases) percentage of infected cells was lower in seven sustained responders (mean: 11.7%, range: 1 24.6%) than in 14 transient responders (mean: 28.2%, range: 7.4-75.5%) and in seven non-responders (mean: 41%, range: 8.8-86%) (P=0.036). At the end of therapy the computed infected cell number was <100 cells/ml of extracellular fluid in all sustained responders (mean: 18.4, range: 1.7-48), in three transient responders (mean: 3500, range: 1.52-17500) and in none non-responders (mean: 28500, range: 1200-96000) (P=0.003). Overall of 10 patients with less than 100 infected cells/ml at the end of therapy seven (six had combination therapy) showed sustained response. All three relapsers received interferon monotherapy. CONCLUSION: The analysis of infected cells dynamics using the new model might be useful to tailor treatment duration in patients with combination therapy. PMID- 14760884 TI - A randomized trial of nelfinavir and abacavir in combination with efavirenz and adefovir dipivoxil in HIV-1-infected persons with virological failure receiving indinavir. PMID- 14760886 TI - Hepatitis B virus genotypes B and C do not affect the antiviral response to lamivudine. AB - To date, there have been no studies examining the role of hepatitis B virus (HBV) genotypes on the response to lamivudine therapy and the development of YMDD mutations. The present study aimed at determining any differences in the antiviral response and risk of YMDD mutations between lamivudine-treated patients with HBV genotype B and genotype C. Eighty-two patients receiving lamivudine were recruited. HBV genotypes at baseline and YMDD mutations at week 52 were determined by line probe assays (LiPA). HBV DNA levels were determined by the Cobas Amplicor HBV Monitor Test. Seventeen (20.7%) and sixty-four (78%) patients had single genotypes of B and C, respectively. At both week 24 and 52 there were no differences in the median reduction of HBV DNA levels (median 4 logs drop), the median reduction of alanine aminotransferase (ALT) levels, and the proportion with normalization of ALT [8/8 (100%) vs 26/37 (70.3%), P=0.19] between patients with genotypes B and C. The rate of HBeAg seroconversion [3/17 (17.6%) vs 6/64 (9.4%), P=0.39] and the chance of YMDD mutation development [3/17 (17.6%) vs 12/64 (18.8%), P=1.0] at week 52 were also similar between patients with genotype B and C, respectively. In conclusion, there was no difference in the antiviral response and the rate of development of YMDD mutations in Chinese patients with genotype B and C after 1 year of lamivudine. Determination of HBV genotypes before lamivudine therapy was probably not an important pretreatment investigation to predict antiviral responses in Chinese patients. PMID- 14760888 TI - Unique keratinocyte-specific effects of interferon-gamma that protect skin from viruses, identified using transcriptional profiling. AB - Interferon (IFN)-gamma, is a multifunctional, immunomodulatory cytokine with cell type-specific antiviral activities, particularly important in skin, where it is implicated in many diseases ranging from warts to psoriasis and cancer. Since epidermis is our first line of defence against many viruses, we investigated the molecular processes regulated by IFN-gamma in keratinocytes using DNA microarrays. We identified the IFN-gamma-regulated keratinocyte-specific genes in keratinocytes, IFN-gamma-induced tight junction proteins, presumably to deny viruses paracellular routes of infection. Furthermore, differing from published data, we find that IFN-gamma suppressed the expression of keratinocytes differentiation markers including desmosomal proteins, cornified envelope components and suprabasal cytokeratins. Inhibition of differentiation may interfere with the epidermal tropism of viruses that require differentiating cells for growth, for example, papillomaviruses. As in other cell types, IFN gamma induced HLA, cell adhesion and proteasome proteins, facilitating leukocyte attraction and antigen-presentation by keratinocytes. IFN-gamma also induced chemokine/cytokines specific for mononuclear cells. IFN-gamma suppressed the expression of over 100 genes responsible for cell cycle, DNA replication and RNA metabolism, thereby shutting down many nuclear processes and denying viruses a healthy cell in which to replicate. Thus, uniquely in keratinocytes, IFN-gamma initiates a well-organized molecular programme boosting host antiviral defences, obstructing viral entry, suppressing cell proliferation and impeding differentiation. PMID- 14760889 TI - Proteasome inhibitors: a novel tool to suppress human cytomegalovirus replication and virus-induced immune modulation. AB - Recently, we like others, demonstrated that systemic inflammation is the most important mechanism involved in (re)activation of human cytomegalovirus (HCMV) in both immunocompetent patients. By in vitro studies the eukaryotic transcription factor NF-kappaB could be identified as the key mediator of TNF-alpha- and IE1 dependent stimulation of the HCMV IE1/2 enhancer/promoter activity, which is crucial for initiation of viral gene expression during reactivation from latency as well as productive infection. The enzymatic proteasome complex plays a central role in regulating intracellular processes, including the activation of NF kappaB. As present antiviral strategies target mainly late events in HCMV replication (DNA replication, virus assembly) that do not completely prevent virus mediated immunopathogenesis, we wondered whether proteasome inhibitors might be a novel tool for targeting the interaction between inflammation and HCMV (re)activation. Here, proteasome inhibitors like MG132, PSI, II and III (MG262) have been shown to block both TNF-alpha-associated up-regulation of the HCMV IE1/2 enhancer/promoter in monocytic cells in an in vitro transient transfection system and HCMV replication in permissive embryonal fibroblasts. Importantly, ganciclovir-resistant HCMV strains are sensitive to proteasome inhibitors. The effect of proteasome inhibitors on HCMV replication was found to be specific as replication of other herpes viruses, like HSV-1 and HSV-2, under identical experimental conditions was not influenced. Inhibition of HCMV replication correlated with a delayed and significantly reduced expression of IE proteins, particularly of the IE2 protein, suggesting that MG132 blocks HCMV replication at an immediate early stage of infection. Early and late protein synthesis as shown exemplary for the pp52 (DNA-binding protein) and p68 (structural protein) protein production and viral DNA synthesis were also inhibited. Suppression of HCMV replication could be correlated with an increased cytosolic accumulation of IkappaB as well as a reduced NF-kappaB binding activity in nuclear extracts of MG132-treated cells, which mainly regards NF-kappaB p50. MG132 also reduced the immune modulatory activity of the virus by abrogating virus-induced up-regulation of cellular ICAM-1. These data suggest that short-term therapy with proteasome inhibitors might be an alternative strategy to prevent (re)activation, replication and immune modulatory activity of HCMV in patients with systemic inflammation. PMID- 14760890 TI - Age, adherence and injection drug use predict virological suppression among men and women enrolled in a population-based antiretroviral drug treatment programme. AB - OBJECTIVES: To characterize 1-year virological response to antiretroviral therapy and its determinants by sex. METHODS: This is a population-based analysis of antiretroviral therapy naive HIV-positive adult men and women. Factors associated with sex and with plasma HIV RNA viral load suppression to below 500 copies/ml were examined using non-parametric tests and logistic regression analyses. RESULTS: A total of 739 subjects (92 women and 647 men) were eligible. Female participants were younger (34 vs 37 years; P < 0.001), less likely to have AIDS (6.5 vs 14.4%; P = 0.039), more frequently injection drug users (44.6 vs 25.2%; P = 0.001) and were less likely to be adherent to therapy (34.8 vs 62.9%; P < 0.001) than male participants. There was no difference in baseline median CD4 count (P = 0.424) or HIV RNA levels (P = 0.140), physician experience (P = 0.057), or with respect to antiretroviral regimens containing protease inhibitors or non-nucleoside reverse transcriptase inhibitors (P = 0.911). With treatment, 46.7% (43/92) of women and 64.8% (419/647) of men (P = 0.001) suppressed HIV RNA viral load to below 500 copies/ml at 1 year. In a multivariate analysis, the association of sex with HIV RNA response to antiretroviral therapy fell from statistical significance (odds ratio 1.18; 95% CI: 0.72-1.95) after adjusting for adherence, injection drug use and age. CONCLUSION: Our data indicate that in this population-based setting, sex differences in 1-year virological response to antiretroviral therapy are explained by age, adherence and injection drug use. PMID- 14760891 TI - Phenotype or virtual phenotype for choosing antiretroviral therapy after failure: a prospective, randomized study. AB - BACKGROUND: Resistance testing is useful in the management of virological failure patients, although the best method to be used in clinical practice has not been determined. METHODS: A prospective, randomized, double-blind, multicentre, controlled clinical trial was performed to compare the usefulness of drug resistance testing with a recombinant viral phenotype method or with a virtual phenotype, a genotyping interpretation system. Planned 300 HIV-infected adults failing their current antiretroviral therapy (HIV RNA > 1000 copies/ml) were centrally randomized 1:1 to resistance testing with a recombinant viral phenotype method or with a virtual phenotype, after stratifying according to previous drug exposure (one or two versus three drug classes). Percent of patients with HIV RNA suppression (% < 400 copies/ml) after 24 weeks was the primary outcome variable. Median HIV RNA concentration and change from baseline in HIV RNA concentration were also used to compare effectiveness. An extended analysis was performed at week 48. RESULTS: Of the 300 patients enrolled, a total of 276 patients could be analysed; 139 patients were randomized to the phenotype group and 137 patients were randomized to the virtual phenotype group. After 24 weeks of follow-up, 46.8 and 56.2% of patients had HIV RNA < 400 copies/ml (P = 0.1) in the phenotype and virtual phenotype, respectively. Mean decrease from baseline in viral load was 1.0 and 1.3 log copies/ml in the phenotype and virtual phenotype groups, respectively (P = 0.017). In a multivariate linear regression analysis, after adjusting for baseline HIV RNA and adherence to treatment, the virtual phenotype was associated with a greater mean decrease in plasma HIV RNA (P = 0.0063). The results observed at week 48 were similar. CONCLUSIONS: Virtual phenotype is at least as effective as phenotype when used to select an optimized treatment for patients who have failed one or more antiretroviral regimens. PMID- 14760892 TI - Impact of early versus late adherence to highly active antiretroviral therapy on immuno-virological response: a 3-year follow-up study. AB - OBJECTIVE: To assess the impact of different patterns of adherence to highly active antiretroviral therapy (HAART), in particular, the relative impact of early and late adherence, on long-term immuno-virological response in HIV infected individuals started on a protease inhibitor-containing regimen. DESIGN: Clinical, immuno-virological and self-reported adherence data were collected at 4 (M4), 12 (M12), 20 (M20), 28 (M28) and 36 (M36) months after HAART initiation in the French APROCO cohort. METHODS: A standardized self-administered questionnaire classified patients as non-adherent, moderately or highly adherent at each visit. Stable viral suppression at both M28 to M36, and a CD4 cell increase > 200 between M0 and M36 were used as outcome measures. RESULTS: Of the 582 patients followed regularly through M36, 360 patients had complete adherence data. Although 59.2% were highly adherent at M4, only 25.8% maintained consistent high adherence throughout the follow-up. High adherence at M4 was independently associated with both stable viral suppression at M28-M36 [OR (95% CI): 2.8 (1.4 5.5)] and a CD4 cell increase > 200 during the same period [OR (95% CI): 3.9 (1.7 9.7)]. However, 'moderately adherent' patients between M12 and M36 had the same likelihood [OR (95% CI): 1.9 (1.1-3.2)] as patients who were always high adherent [OR (95% CI): 1.9 (1.1-3.2)] of achieving stable viral load suppression, relative to those who reported non-adherence episodes. CONCLUSION: Optimizing adherence in the early months of treatment is crucial to ensure long-term immuno-virological high adherence during follow-up have a less negative impact. Priority should be given to interventions aimed to improve adherence in the early months of HAART. PMID- 14760895 TI - The effect of number of mutations and of drug-class sparing on virological response to salvage genotype-guided antiretroviral therapy. AB - OBJECTIVE: To assess on longitudinal data the impact of number of drug-associated mutations at genotype resistance testing (GRT) and history of previous exposure to antiretrovirals on the virological response to genotype-guided antiretroviral therapy. METHODS: Subjects that failed HAART who underwent GRT between June 1999 and March 2002 were enrolled. GRT was performed by Viroseq-2 with expert advice offered to physicians. Main outcome was reaching undetectable (< 80 copies/ml) HIV-1 RNA level after GRT and maintaining undetectable viraemia for at least 6 months. The number of mutations conferring resistance to each class of antiretrovirals was categorized and their effect on virological outcome investigated. Mutations considered in the analysis were those reported by the IAS USA in 2002. Multivariate analysis was performed by Cox proportional hazard model. RESULTS: Four-hundred-and-seventy consecutive subjects were enrolled and followed-up for a median of 14 (IQR 9-19) months after GRT. Sustained undetectable viraemia was reached by 80 of 449 subjects (18%). Using as end-point reaching and maintaining for at least 6 months < 400 copies/ml after GRT, 103 out of 447 subjects (23%) reached the outcome. For each single protease inhibitor (PI)-, nucleoside reverse transcriptase inhibitor (NRTI)-and non-nucleoside reverse transcriptase inhibitor (NNRTI)-associated mutation, there was a reduction of, respectively, 11% (P = 0.008), 12% (P = 0.001) and 39% (P = 0.005) in the likelihood of reaching virological outcome. Subjects carrying > or = 6 mutations to NRTIs, > or = 7 mutations to PIs and > or = 2 mutations to NNRTIs were less likely to reach the virological success compared with those carrying 0 1 (NRTI and PI) or 0 (NNRTI) mutations [HR = 0.25 (95% CI: 0.10-0.65); HR = 0.33 (950% CI: 0.16-0.67); HR = 0.33 (95% CI: 0.14-0.77)], respectively. However, at multivariate analysis the probability of reaching a favourable virological outcome in patients with > or = 7 mutations to PIs, if naive for NNRTIs [HR = 1.74 (0.69-4.36)], and in subjects with > or = 2 mutations for NNRTIs if naive for PIs [HR = 1.23 (0.22-6.80)], was comparable to those observed in patients with none or one mutation. CONCLUSIONS: Our data showed a non-linear association between resistance-conferring mutations and virological outcome. GRT-guided therapy still provided remarkable chances of durable virological success even in subjects with > or = 7 mutations to PIs and in subjects with > or = 2 mutations to NNRTIs, when the subjects did not have a three-class exposure or if GRT showed no evidence of mutations for a drug class. GRT and as-long-as-possible sparing of a drug class could be a convenient strategy for long-term management of drug failing patients. PMID- 14760894 TI - Efficacy and safety of ritonavir/indinavir 100/400 mg twice daily in combination with two nucleoside analogues in antiretroviral treatment-naive HIV-infected individuals. AB - OBJECTIVE: To evaluate the efficacy and tolerability of indinavir/ritonavir (IDV/RTV) 400/100 mg twice daily in combination with two nucleoside reverse transcriptase inhibitors in antiretroviral-naive patients. DESIGN AND METHODS: Antiviral therapy-naive patients with plasma HIV-1 RNA > 5000 copies/ml were enrolled in this pilot, single-arm study. CD4 cell count and viral load were evaluated at weeks (W) 4, 12, 24 and every 3 months until W48. The primary end point was the percentage (%) of patients with viral load < 400 copies/ml at W48. Intent-to-treat (ITT) (missing values or change in treatment equalled failure) and on-treatment (OT) analyses were performed. RESULTS: Forty patients were enrolled. Baseline median viral load was 5.36 log10 copies/ml, median CD4 count was 84 cells/mm3. At W48 by ITT analysis, the % patients with viral load < 400 copies/ml was 65% (95% CI: 48-79) and 50% (95% CI: 35-65) with viral load < 50 copies/ml, and 96% (26/27) (95% CI: 89-100) and 74% (95% CI: 57-91], respectively, by OT analysis. The median decrease in viral load at W48 was -3.83 log10 copies/ml (-0.1; -5.19) and the median increase in CD4 was +167 cells/mm3 (6-474 cell/mm3). At W4 (34/40), the median IDV C(min) was 500 ng/ml (range 5 8100) with 91% of patients with an adequate IDV C(min) > 150 ng/ml. Ten patients discontinued the study treatment before W48: adverse events (eight), patient's will (one) and simplification of therapy (one). Three patients were lost to follow-up. Only one virological failure occurred and was associated with poor compliance and sub-optimal concentrations of IDV/RTV. CONCLUSIONS: IDV/RTV 400/100 mg twice daily is an effective and safe first-line antiretroviral therapy. The simplicity and the low cost of IDV/RTV is of major interest particularly in countries with limited resources. PMID- 14760896 TI - Contribution of nucleoside-analogue reverse transcriptase inhibitor therapy to lipoatrophy from the population to the cellular level. AB - OBJECTIVES: It has been proposed that the contribution of nucleoside-analogue reverse transcriptase inhibitor (NRTI) therapy to subcutaneous fat wasting involves adipose tissue-specific mitochondrial DNA toxicity. We have investigated the relationships between NRTI therapy, adipocyte mitochondrial DNA content, evidence of toxicity in adipose tissue and fat wasting in Caucasian male Western Australian HIV Cohort study participants. METHODS: Longitudinal mixed effects analysis of fat wasting was undertaken in individuals receiving initial stavudine or zidovudine-containing highly active antiretroviral measurements). Adipocyte mitochondrial DNA (mtDNA) depletion was also assessed according to current NRTI therapy in 92 subcutaneous fat biopsies from 69 HIV-positive individuals and seven healthy controls, and results were correlated with fat wasting among a subset of patients with biopsy data receiving initial stavudine- or zidovudine containing HAART (n = 22, 103 DEXA measurements). Confocal microscopy was performed in 22 obtained after initiating/switching NRTI therapy. RESULTS: Stavudine therapy was associated with more severe adipocyte mitochondrial DNA depletion (P < 0.001) and fat wasting over time (P = 0.002) compared with zidovudine therapy in independent analyses. Among patients with concurrent biopsy and longitudinal DEXA data, fat wasting was associated with duration of NRTI therapy (P = 0.001) and adipocyte mtDNA copies/cell (P = 0.01). In this analysis, the significant association between choice of stavudine versus zidovudine and fat wasting (P = 0.03) was lost after adjustment for the effect of mtDNA depletion (P = 0.13). Confocal analysis provided direct evidence of a relationship between severity of adipose tissue toxicity and mitochondrial DNA depletion. No significant effects of HIV protease inhibitor therapy were detected in these analyses. CONCLUSIONS: Severity of subcutaneous fat wasting is primarily determined by choice of NRTI therapy (stavudine versus zidovudine) and by duration of exposure to the relevant NRTI. At the cellular level, evidence is provided that this effect manifests through NRTI-induced mitochondrial DNA depletion. PMID- 14760897 TI - Monitoring the emergence of hepatitis B virus polymerase gene variants during lamivudine therapy in human immunodeficiency virus coinfected patients: performance of CLIP sequencing and line probe assay. AB - Sera from 12 patients infected with human immunodeficiency virus and hepatitis B virus (HBV), on lamivudine as part of an antiretroviral therapy, were retrospectively analysed for the presence of HBV polymerase mutations by the line probe assay, INNO-LiPA HBV DR, and by the direct sequencing assay, TRUGENE HBV genotyping kit. Results at codons 180, 204 and 207 were compared for 44 samples. Full concordance was observed for 81.4% of the 129 analysed codons. Discordance involved only mixed populations: LiPA detected additional species in 19 codons and TRUGENE in five. Viral breakthrough occurred in seven patients, 12-33 months after lamivudine initiation. In five cases with close sampling available, both assays detected mutations before the rise in viral load, although earlier by LiPA for three patients. The time interval between the first mutant detection and viral escape ranged from 2 to 22 months. Mutations were detected in four of the five remaining patients: 1) at therapy initiation in a primary non-responder; 2) after 37 months, but replication became undetectable after tenofovir introduction; 3) transiently at 6 months by LiPA but treatment was ceased thereafter; 4) after 23 months but replication levels remained low during a 5 year follow-up. Interestingly, TRUGENE sequencing identified on late samples from three patients a variant carrying rtV173L plus rtL180M plus M204V mutations, having the in vitro characteristics of 'vaccine escape' mutants. Both assays appear to be valuable tools for the early detection of mutated HBV strains. The detection of genotypic therapeutic decision-making, although clinical or other virological factors may determine the rapidity of the viral breakthrough. PMID- 14760898 TI - Predictors of tonsillar tissue HIV-1 viral burden at baseline and after 1 year of antiretroviral therapy. AB - In 81 antiretroviral-navie HIV-1 chronic-infected patients, we found a correlation among tonsillar tissue viral load, and virological and immunological measures in blood at baseline. No correlation was observed after 1 year of antiretroviral therapy. A protease inhibitor-containing regimen was the best predictor of good tonsillar tissue virological response. PMID- 14760893 TI - A randomized trial comparing initial HAART regimens of nelfinavir/nevirapine and ritonavir/saquinavir in combination with two nucleoside reverse transcriptase inhibitors. AB - BACKGROUND: A triple-class HAART regimen may be associated with a better virological effect than conventional regimens, but may also lead to toxicity and more profound resistance. METHODS: Randomized, controlled, open-label trial of 233 protease inhibitor- and non-nucleoside reverse transcriptase inhibitor-naive HIV-infected patients allocated to a regimen of nelfinavir and nevirapine (1250/200 mg twice daily; n = 118) or ritonavir and saquinavir (400/400 mg twice daily; n = 115), both in combination with two nucleoside reverse transcriptase inhibitors. The primary end-point was HIV RNA < or = 20 copies/ml after 48 weeks (missing value = failure). Patients remained under follow-up also in case of switch from the randomized therapy. RESULTS: At baseline, the median CD4 cell counts were 126 (range: 0-942) (nelfinavir/nevirapine) and 150 (0-642) (ritonavir/saquinavir) cells/mm3, and HIV RNA measurements 5.0 copies/ml (1.3 6.4) in both groups. A total of 102 (86%) and 101 (88%) were antiretroviral naive. 44% discontinued randomized therapy; P = 0.13. Of these, 80 and 73% switched therapy due to adverse events; P = 0.99. At week 48, 69 and 56%, respectively, had a HIV RNA < or = 20 copies/ml; P = 0.037. CONCLUSION: A regimen of nelfinavir/nevirapine had a favourable virological effect and tolerability over a 48-week period compared with ritonavir/saquinavir, when administered in combination with two nucleoside reverse transcriptase inhibitors. However, more extensive follow-up is required to determine the long-term consequences of triple class HAART regimens, including the development of broad drug resistance. PMID- 14760900 TI - Anatomy of the pulp-chamber floor. AB - Locating the number and position of orifices on pulp-chamber floors can be difficult. This is especially true when the tooth being treated is heavily restored, malposed, or calcified. After evaluating 500 pulp chambers of extracted teeth, new laws for finding pulp chambers and root-canal orifices are proposed. The use of these laws can aid in the determination of the pulp-chamber position and the exact location and number of root canals in any individual tooth. PMID- 14760899 TI - The outcome of endodontic retreatment: a 2-yr follow-up. AB - Retreatment is common in endodontics. The purpose of this article was to classify the different clinical situations encountered in retreatment cases and relate them to the outcome after an observation period of 24 months. A total of 425 patients (452 teeth) from 451 patients, consecutively admitted for root-canal retreatment, were monitored during a 24-month period. All teeth (254 molars, 107 premolars, and 91 single-root anterior teeth) were divided into two major categories: teeth with modified anatomy from previous endodontic treatment (root canal-morphology altered) and teeth in which no significant anatomical changes were made by the former endodontic treatment (root-canal-morphology respected). Although the overall success was 69.03%, the success in the root-canal-morphology respected group was 86.8% and in the root-canal-morphology-altered group 47% (Mann-Whitney U test p < 0.0001). The clinical success of an endodontic retreatment seems to depend on whether alterations in the natural course of the root canals were caused by previous root-canal treatment. PMID- 14760903 TI - Attachment and morphological behavior of human periodontal ligament fibroblasts to mineral trioxide aggregate: a scanning electron microscope study. AB - The attachment and morphology of human periodontal ligament fibroblasts to mineral trioxide aggregate (MTA) was evaluated using a scanning electron microscope. The material was placed at an apical cavity of 30 single-rooted slices of extracted human teeth. The specimens were divided into two groups of 15 root slices each (freshly mixed and set state). For each experimental group, five root slices were used per observation period (4, 8, and 24 h). A set of two glass slides was used per observation period for the control group. The experiments were performed in tissue-culture cluster 96-well plates in which 1 ml of human periodontal ligament fibroblast cell suspension was placed over the MTA filling and the control glass slides. For the positive-control group, 0.5 ml of methyl methacrylate 2% (vol/vol) was added to the cell suspensions before being dispensed into the wells. Results showed the normal cell morphology in the negative controls. Few round cells with less smooth surfaces and many rough blebs were seen in the positive control, and most of these cells did not show any attachment to the substratum. Similar observations were seen with the freshly prepared-MTA group. In the set-MTA group, cells were round and flattened, displayed smooth surfaces, and appeared to be tightly attached to MTA. It was concluded that the quality and quantity of cell attachment to the retrofilling material could be used as a criterion to evaluate material's toxicity. This research (FN#1077) is registered with the College of Dentistry research center, King Saud University, Riyadh, Saudi Arabia. The author thanks the administration of the King Faisal Specialist Hospital and Research Center, Riyadh, Saudi Arabia, and in particular Dr. M. N. Al-Ahdal for providing the use of the Molecular Virology and Infectious Disease Laboratory, Mr. Yunus Siddiqui for his support, and Dr. Saad AL-Nazhan for his assistance in preparing the manuscript. PMID- 14760902 TI - Hard-tissue healing after application of fresh or set MTA as root-end-filling material. AB - The purpose of this study was to compare the effect of fresh mineral trioxide aggregate (MTA) with set MTA on hard-tissue healing after periradicular surgery. The root canals of 24 mandibular premolars in four 2-yr-old beagle dogs were filled with MTA. Two weeks later the root ends of half of the samples were surgically exposed and resected to the set MTA within the canals. After exposing and resecting the other 12 root ends, they were prepared with ultrasonic instrumentation and preparations were filled with fresh MTA. After 4 months, the animals were killed. Hard-tissue healing was analyzed histomorphometrically. The results indicated that although freshly placed MTA resulted in a significantly higher incidence of cementum formation (12 of 12 versus 8 of 12, p = 0.028), there is no significant difference in the quantity of cementum or osseous healing associated with freshly placed or set MTA when used as root-end-filling material. PMID- 14760901 TI - The effect of calcium sulfate on hard-tissue healing after periradicular surgery. AB - The purpose of this study was to determine the effect of calcium sulfate (CS) on cementum deposition and osseous healing after periradicular surgery. The root canals of 24 mandibular premolars in four 2-yr-old beagle dogs were endodontically treated, followed 2 weeks later by periradicular surgery. Mineral trioxide aggregate (MTA) was used as root-end-filling material. The right or left side was assigned at random to receive CS alpha-hemihydrate or no material in the osteotomy sites before closure. The animals were killed after 4 months. Hard tissue healing was analyzed histomorphometrically. All samples displayed evidence of cementum deposition adjacent to the root-end fillings and bone regeneration in the osteotomy sites. The data was analyzed using the Mann-Whitney U test, comparing the scores for cementum and osseous healing of the two groups at significance level of alpha = 0.05. The results indicated that placement of CS in osteotomy sites after periradicular surgery does not significantly affect periradicular healing. PMID- 14760905 TI - Comparative study of sealing ability of two root-end-filling materials with and without the use of dentin-bonding agents. AB - The purpose of this study was to examine microleakage of two root-end-filling materials with and without the use of bonding agents. For microleakage measurement a fluid transport model was used. Sixty single-rooted teeth were used in this study. They were prepared with step-back technique and then an apicoectomy was performed. Then the teeth were divided into four groups. Group A were filled with Fuji II LC glass-ionomer cement, group B were filled with Fuji II LC and a new bonding agent, Fuji Bond, group C were filled with Admira composite resin, and group D were filled with Admira and Admira Bond, a new bonding agent. At 24 h, 1 month, and 2 months after filling the leakage along these filling materials was determined under a low pressure of 0.1 atm. At all experimental times, glass-ionomer groups showed significantly less microleakage than resins groups. Between Admira and Admira Bond groups, significantly less leakage was observed in the root sections with Admira Bond at 24 h. PMID- 14760904 TI - Diffusion of Ca(OH)2 associated with different vehicles: chromatographic study (high-performance liquid chromatography). AB - Using high-performance liquid chromatography (HPLC), small amounts of liquid samples in which 25 premolar human teeth were immersed were evaluated. Each tooth was immersed separately in 800-ml flasks with distilled ultra-pure deionized water and remained there for 1678 h after the filling of their canals with Ca(OH)2 associated with different vehicles: group 1: polyethylene glycol and colophon (Calen); group 2: glycerin and camphorated paramonochlorophenol; group 3: camphorated paramonochlorophenol; group 4: glycerin and tricresol formol; and group 5: anesthetic solution (Citanest). Five polyethylene tubes were filled with each of these pastes and placed unsealed in similar flasks. At the end of this period, HPLC analyses of the aqueous medium related to each group were performed to detect other substances that had diffused from the pastes used in the canals of the teeth other than calcium and hydroxyl ions. Although the groups presented different maximum peaks when there was no barrier, they all showed higher values than when the tooth was present. At least 15 substances other than Ca2+ and OH- were detected in the aqueous medium of group 4. Analyzing the HPLC graphs, we concluded that not only Ca2+ and OH-, but also a considerable quantity of other components of the pastes diffused through the dentine and reached the external root surface. PMID- 14760906 TI - A comparative study of crack development in stainless-steel hedstrom files used with step-back or crown-down techniques. AB - The purpose of this study was to evaluate the effect of step-back (SB) and crown down (CD) techniques on crack development of stainless-steel Hedstrom files (H files). Ninety-seven mesial root canals of freshly extracted human mandibular molars were selected. Six sets of H-files (#15, #20, and #25 ISO size each) were used with SB and CD techniques for the chemomechanical preparation of 8, 16, and 24 root canals using 2.5% NaOCl as an irrigant and EDTA gel as a chelating agent. Files were sterilized in an autoclave between successive uses. A seventh unused set served as control. After macroscopic examination all files were embedded in epoxy resin, ground, polished, and studied under a metallographic microscope. The maximum crack size of each file was measured by means of standard image-analysis procedures. Macroscopically the files presented no signs of plastic deformation, apart from a file #15, used with SB technique, which fractured after 16 root canal preparations. The microscopic examination showed no cracks: on control files, on files used with the CD technique for 8 and 16 cycles, on file #20 used with the SB technique for 8 and 16 cycles, as well as on file 25 used with the SB technique for 8 cycles. Cracks were found on H-file #15 used with the SB technique for 8 and 16 cycles, on file 25 used with the SB technique for 16 cycles, and on all files used for 24 cycles with both SB and CD techniques. Under the conditions of this study it was concluded that the instrumentation technique is deeply implicated in the crack development. Crack propagation was accelerated when SB technique was used. PMID- 14760908 TI - Effects of the hydroxyl radical and hydrogen peroxide on tooth bleaching. AB - The mechanisms of bleaching of discolored coronal teeth using hydrogen peroxide (H2O2) were investigated. In a scanning-electron-microscopy study, the intertubular dentin and peritubular dentin were dissolved by high concentrations of H2O2, which is used for bleaching. The X-ray diffraction study showed that hydroxyapatite was not influenced by H2O2. In an electron-spin-resonance study, more hydroxyl radical (* OH) was detected as the H2O2 concentration was increased. When amino acids that are core components of dentin proteins, such as proline and alanine, were added to H2O2, the generation of * OH decreased, but there was no change when glycine was added. A nuclear-magnetic-resonance study showed that proline was degraded completely by H2O2, the structure of alanine changed slightly, and glycine was not affected by H2O2. It is suggested that H2O2 and * OH do not influence the inorganic tissue of dentin but attack the organic component of dentin. These facts suggest that * OH has the main role in tooth bleaching with H2O2. PMID- 14760907 TI - Microorganisms growth in endodontic citric-acid solutions with and without microbiological stabilizer. AB - The purpose of this study was to determine if citric-acid solutions for endodontic use can become contaminated and if the addition of a microbiological stabilizer to the acid would modify this situation. Five types of microorganisms were inoculated in tubes containing 10% citric-acid solutions, pH 1.8 (group A1), and 10% solution, pH 1.8, combined with 0.1% sodium benzoate (group A2). The results demonstrated that solutions of group A1 were contaminated with Candida albicans in 100% of the tubes, Escherichia coli in 80%, and Enterococcus faecalis in 50%. When stabilizer was added (group A2), C. albicans grew in only 30% of tubes and all bacterial species were neutralized. PMID- 14760909 TI - Microhardness of packable composites used as posts condensed with different methods. AB - The purpose of this in vitro study was to evaluate the microhardness of packable composite compacted with hand or ultrasonic pluggers in post spaces. A total of 168 extracted human anterior teeth were prepared with Gates Glidden and ParaPost drills after obturation. A primer and a dentin-bonding agent were applied to the etched surface. Half of the specimens were compacted with an ultrasonic tip and the other half with a mechanical hand compactor and all specimens were polymerized for each increment. The roots were then sectioned horizontally from 0, 2.5, 3, 3.5, 4, 4.5, and 5 mm starting from the coronal and sliced 2.5-mm long. The microhardness test was applied to each specimen. There were statistically significant differences between ultrasonically and hand-condensed groups in whole specimens (p < 0.001). There were no significant differences between groups of 0, 2.5, and 3 mm until 3.5 mm was reached. After 3.5-mm depth, there were significant differences between the groups (p < 0.001). Ultrasonic condensation of packable composites provided mechanical advantage over hand condensation in root canals measuring microhardness in different depths. However, when microhardness was measured at different depths, values decreased after the depth of 3.5 mm in both condensation groups. PMID- 14760910 TI - Effect of gutta-percha solvents on mineral contents of human root dentin using ICP-AES technique. AB - The purpose of this in vitro study was to evaluate the mineral contents of root canal dentin before and after treatment with two commonly used gutta-percha solvents: chloroform and halothane. Twenty extracted human premolars, whose crowns and apical thirds had been removed, were used. Pulp tissues were removed and the teeth were randomly divided into two groups including 10 teeth each. Root canals were enlarged with Gates Glidden burs (#1, 2, and 3). Dentin chips were obtained and saved in plates to serve as a control. Root-canal-dentin walls were then treated with chloroform or halothane for 15 min. Dentin chips were again obtained using Gates Glidden burs (#4, 5, and 6). The levels of five elements, calcium, phosphorus, magnesium, potassium, and sulfur, in each specimen were analyzed using ICP-AES (inductively coupled plasma atomic emission spectrometry) technique. Changes in the levels of the chemical elements were recorded. Differences between the groups were statistically analyzed using Mann-Whitney U test. There was a significant decrease in Ca level and significant increase in Mg level after treatment with halothane (p < 0.05). There was a significant increase in Mg level after treatment with chloroform. The changes in other elements levels after treatment with gutta-percha solvents were minimal and statistically not significant (p > 0.05). As a result it was concluded that gutta-percha solvents have effect on mineral contents of root dentin. PMID- 14760914 TI - One-dimensional rabbit sinoatrial node models: benefits and limitations. AB - INTRODUCTION: Cardiac multicellular modeling has traditionally focused on ventricular electromechanics. More recently, models of the atria have started to emerge, and there is much interest in addressing sinoatrial node structure and function. METHODS AND RESULTS: We implemented a variety of one-dimensional sinoatrial models consisting of descriptions of central, transitional, and peripheral sinoatrial node cells, as well as rabbit or human atrial cells. These one-dimensional models were implemented using CMISS on an SGI Origin 2000 supercomputer. Intercellular coupling parameters recorded in experimental studies on sinoatrial node and atrial cell-pairs under-represent the electrotonic interactions that any cardiomyocyte would have in a multidimensional setting. Unsurprisingly, cell-to-cell coupling had to be scaled-up (by a factor of 5) in order to obtain a stable leading pacemaker site in the sinoatrial node center. Further critical parameters include the gradual increase in intercellular coupling from sinoatrial node center to periphery, and the presence of electrotonic interaction with atrial cells. Interestingly, the electrotonic effect of the atrium on sinoatrial node periphery is best described as opposing depolarization, rather than necessarily hyperpolarizing, as often assumed. CONCLUSION: Multicellular one-dimensional models of sinoatrial node and atrium can provide useful insight into the origin and spread of normal cardiac excitation. They require larger than "physiologic" intercellular conductivities in order to make up for a lack of "anatomical" spatial scaling. Multicellular models for more in-depth quantitative studies will require more realistic anatomico-physiologic properties. PMID- 14760912 TI - Predictable Therma-fil removal technique using the system-B heat source. AB - A clinical tip is suggested to assist in the removal of Therma-fil obturators during conventional endodontic retreatment. Using a heat source such as the System-B, the plastic carriers that are normally an obstacle to retreatment can be efficiently removed using the technique described. PMID- 14760913 TI - Should we or shouldn't we? PMID- 14760915 TI - Mechanisms of termination of atrial fibrillation by Class I antiarrhythmic drugs: evidence from clinical, experimental, and mathematical modeling studies. AB - Sodium channel blocking drugs (Class I antiarrhythmic agents) have been used for the termination of atrial fibrillation (AF) and for sinus rhythm maintenance for almost 100 years. Despite this long history, the mechanisms that underlie their efficacy in AF remain poorly understood. Classic notions about the determinants of cardiac reentry, as embodied in leading circle theory, and of AF, as reflected in the multiple wavelet hypothesis, suggest that cardiac conduction slowing should promote, rather than prevent, AF. This article reviews the evidence (both clinical and experimental) for the efficacy and mechanisms of action of Class I antiarrhythmic agents in AF. Application of mathematical models of AF to the evaluation of Class I mechanisms is discussed, and recent insights into the latter are presented. A better understanding of the ways in which Na+ channel blockers affect AF will be useful, not only for new antiarrhythmic drug development but also for gaining insight into the mechanisms of the arrhythmia. PMID- 14760911 TI - In vitro antibacterial activities of root-canal sealers by using two different methods. AB - The purpose of this in vitro study was to evaluate the antibacterial activity of five different root-canal sealers (RoekoSeal, Ketac-Endo, AH Plus, Sealapex, Sultan). With the use of Enterococcus faecalis as a test organism, both the agar diffusion test (ADT) and direct-contact test (DCT) were performed. For DCT, sealers were mixed and placed on the sidewall of microtiter plate wells. A 10 microl bacterial suspension was placed on the tested material samples. Bacteria were allowed to directly contact to the sealers for 1 h at 37 degrees C. Bacterial growth was then spectrophotometrically measured through every 30 min for 19 h by using an Anthos Labtec HT 2. For ADT, a 200-microl bacterial suspension was spread on brain-heart infusion agar plates. Freshly mixed sealers were poured into uniform wells punched in the agar. After periods of incubation at 37 degrees C for 24 h and 7 days in humid atmosphere, the zones of inhibition of bacterial growth on agar plates were observed and measured. Ketac-Endo, Sultan, and AH Plus had similar results for DCT. These sealers were more potent bacterial-growth inhibitors than Sealapex and RoekoSeal. According to ADT, RoekoSeal showed no antibacterial activity. There was no significant difference among AH Plus, Sealapex, and Sultan (p > 0.05). Ketac-Endo demonstrated lower antimicrobial activity than these sealers (p < 0.05). Time had no effect on the antibacterial activity of the tested sealers (p > 0.05). The antibacterial efficiency of the materials varied according to the tests used. It was concluded that the technique, time, and ingredients of the tested material can affect the results of the microbiological studies. PMID- 14760916 TI - The electrical restitution curve revisited: steep or flat slope--which is better? AB - The electrical restitution curve (ERC) traditionally describes the recovery of action potential duration (APD) as a function of the interbeat interval or, more correctly, the diastolic interval (DI). Often overlooked in modeling studies, the normal ventricular ERC is triphasic, starting with a steep initial recovery at the shortest DIs, a transient decline, and a final asymptotic rise to a plateau phase reached at long DIs. Recent studies have proposed that it would be advantageous to lower the slope of the ERC by drug intervention, as this might reduce the potential for electrical alternans and ventricular fibrillation. This review discusses the pros and cons of a flat versus steep slope of the ERC and draws attention to mechanisms thatjustify the (physiologically) steep slope, rather than a flat slope, as a better design against arrhythmias. Five potential mechanisms are discussed, which allows for a different interpretation of the effect of the slope on arrhythmogenicity. The most important appears to be the physiologic rate adaptive shortening of APD that, by reciprocal lengthening of the DI, allows the subsequent APD to move more quickly from the steep initial ERC phase onto the flat phase. A less steep initial ERC phase would protract the transition toward more fully recovered APD and, in fact, may perpetuate electrical alternans. The triphasic ERC time course in normal myocardium cannot be explained by or fitted to single exponentials or single ion channel recovery kinetics. A simple tri-ionic model is suggested that may help explain the shape of the ERC at various repolarization levels and place APD recovery into perspective with intracellular calcium recycling and recovery of contractile force. PMID- 14760917 TI - Left atrial conduction along the coronary sinus during ectopic atrial tachycardia and atrial fibrillation: a study using correlation function analysis. AB - INTRODUCTION: Correlation function analysis was applied to endocardial electrograms to investigate conduction patterns along the coronary sinus (CS) during sinus rhythm (SR) and atrial tachycardias. METHODS AND RESULTS: Eighteen recordings were obtained from 14 patients with supraventricular tachycardias. Five atrial fibrillation (AF) recordings were compared to 10 SR recordings and 3 ectopic atrial tachycardia (EAT) recordings. The maximum correlation coefficient was used to assess similarity between signals, i.e., if they originate from the same wavefront. The cumulative time delay, calculated as pairwise summation of interelectrode time delays, was used as an indicator of activation sequence along the CS. Method validation using SR showed right-to-left conduction with high correlations in 8 of 10 recordings indicating one single wavefront. EAT recordings showed consistent left-to-right conduction with left atrial foci and right-to-left with right atrial focus and lower correlations than SR. All 5 AF recordings showed predominantly left-to-right conduction direction, also with correlations lower than SR. CONCLUSION: (1) Correlation function analysis can be used to assess agreement between signals and direction of activation spread. (2) Due to the position of CS, the results can be used to derive mechanisms of interatrial conduction. (3) Consistency in electrical activity propagation along CS is common in AF. PMID- 14760919 TI - Echocardiographic and electrocardiographic predictors for atrial fibrillation recurrence following cardioversion. AB - INTRODUCTION: Identification of suitable candidates for cardioversion currently is not based on individual electrical and mechanical atrial remodeling. Therefore, this study analyzed the meaning of atrial fibrillatory rate obtained from the surface ECG (as a measure of electrical remodeling) and left atrial size (as measure of mechanical remodeling) for prediction of early atrial fibrillation (AF) recurrence following cardioversion. METHODS AND RESULTS: Forty-four consecutive patients (26 men and 18 women, mean age 62 +/- 11 years, no antiarrhythmic medication at baseline) with persistent AF were studied. Fibrillatory rate was obtained from high-gain, high-resolution surface ECG using digital signal processing (filtering, QRST subtraction, Fourier analysis) before electrical cardioversion. Univariate and multivariate regression analysis revealed larger systolic left atrial area (Beta = 0.176, P = 0.031) obtained by precardioversion echocardiogram from the apical four-chamber view and higher atrial fibrillatory rate (Beta = 0.029, P = 0.021) to be independent predictors for AF recurrence (n = 13). Stratification based on the regression equation (electromechanical index [EMI] = 0.176 systolic left atrial area + 0.029 fibrillatory rate - 17.674) allowed identification of groups at low, intermediate, or high risk. No patient with an EMI < -1.85 had early AF recurrence, as opposed to 78% with an EMI > -0.25. Intermediate results (40% recurrence rate) were obtained when the calculated EMI ranged between -1.85 and 0.25 (P < 0.001). CONCLUSION: Fibrillatory rate obtained from the surface ECG and systolic left atrial area obtained by echocardiography may predict early AF recurrence in patients with persistent AF. These parameters might be useful in identifying candidates with a high likelihood of remaining in sinus rhythm after cardioversion. PMID- 14760920 TI - Atrial excitation assuming uniform propagation. AB - INTRODUCTION: We investigated the spread of the excitation wave over the atria following initiation in a given focus in an atrial model containing its overall geometry only, i.e., without atrial bundles. METHODS AND RESULTS: The propagation velocity of the excitation wave was taken to be uniform, and the wall thickness was discarded. The timing of excitation of any point on the atrium thus becomes directly proportional to its shortest distance over the atrial wall to the focus. Despite these gross simplifications, the general nature of the excitation sequence found corresponded closely to clinical data reported in the literature. This suggests that the complex overall geometry of the atria dominates the timing of the excitation. A highly intriguing observation from this study was that, when looking at the pathways from the sinus node to all other points on the atrium, prominent routes became visible even though no such pathways formed part of the model of the atrial geometry used. The locations of these prominent routes coincide with those of various distinct bundles in the atria. Possible inferences of these observations are discussed. CONCLUSION: Based upon comparison with data from other studies, it is concluded that, during stable heart rhythms, propagation of the atrial excitation wave is well approximated by an assumption of uniform velocity, even though no atrial bundles were included in the model. The overall geometry seems to be the dominant factor in the spread of excitation. PMID- 14760922 TI - The dominant T wave and its significance. AB - INTRODUCTION: The shapes of the T waves as observed in different leads placed on the thorax are very similar. The dominant T wave is introduced as a means to characterize this general signal shape. Its relationship to the transmembrane potentials of cardiac myocytes is discussed. METHODS AND RESULTS: The source description of a biophysical model that previously was shown to yield realistic T waveforms was analyzed in order to exploit its relation to the transmembrane potentials of the cardiac myocytes at the surface bounding the myocardium. The product of this analysis is the dominant T wave: a waveform that describes the slope of the transmembrane potential. It is shown that the dominant T wave can be estimated easily from the matrix of sampled lead potentials. The timing of its peak reveals the mean of the repolarization times of the involved transmembrane potentials. The amplitude of the peak is the maximum downward slope of the transmembrane potential. This amplitude is independent of the volume conductor effects of the tissues surrounding the heart. The estimate of the dominant T wave retains this property. CONCLUSION: The dominant T wave reflects the derivative of the recovery phase of a generalized transmembrane potential. Its amplitude is independent of the volume conductor properties of the tissues surrounding the heart. This is a unique feature that greatly facilitates the interpretation and application of the other signal features of the dominant T wave. PMID- 14760918 TI - Frequency measures obtained from the surface electrocardiogram in atrial fibrillation research and clinical decision-making. AB - INTRODUCTION: Frequency analysis of fibrillation (FAF) and time-frequency analysis (TFA) were developed recently in order to quantify atrial electrical remodeling in atrial fibrillation (AF) from the surface ECG. This article describes the experience with these two different frequency analysis techniques in consecutive AF patients and discusses possible applications in AF research and clinical decision-making. METHODS AND RESULTS: Baseline 2-minute, high-gain, high resolution ECG recordings using three bipolar leads were obtained from 80 consecutive patients with AF lasting > 24 hours. A power spectrum was obtained using Fourier analysis following spatiotemporal QRST cancellation. The dominant fibrillatory rate (in fibrillations per minute [fpm]) was derived (FAF). Stability of the instantaneous fibrillatory rate measured in overlapping 1-second segments was expressed as the segment proportion with consecutive rate differences < 6 fpm (TFA). An adequate power spectrum that could be submitted for determination of fibrillatory rate was obtained in all patients. Dominant atrial rates ranged between 288 and 534 fpm and showed a high correlation (R = 0. 878 0.911, P < 0.001) when assessed from the three different leads. The average instantaneous fibrillatory rate was inversely related with its stability (R = 0.417, P < 0.001). It was closely related with the dominant fibrillatory rate obtained from FAF (R = 0.948, P < 0.001). A literature review revealed that pharmacologic or electrical cardioversion and AF pace termination success rates were highly dependent on fibrillatory rate. CONCLUSION: Atrial fibrillatory rate and its variability can be reliable obtained from the surface ECG in AF patients. These parameters exhibit a significant interindividual variability allowing individual quantification of the atrial electrical remodeling process and might prove useful for predicting therapy efficacy. PMID- 14760923 TI - Timing of depolarization and contraction in the paced canine left ventricle: model and experiment. AB - INTRODUCTION: For efficient pump function, contraction of the heart should be as synchronous as possible. Ventricular pacing induces asynchrony of depolarization and contraction. The degree of asynchrony depends on the position of the pacing electrode. The aim of this study was to extend an existing numerical model of electromechanics in the left ventricle (LV) to the application of ventricular pacing. With the model, the relation between pacing site and patterns of depolarization and contraction was investigated. METHODS AND RESULTS: The LV was approximated by a thick-walled ellipsoid with a realistic myofiber orientation. Propagation of the depolarization wave was described by the eikonal-diffusion equation, in which five parameters play a role: myocardial and subendocardial velocity of wave propagation along the myofiber cm and ce; myocardial and subendocardial anisotropy am and ae; and parameter k, describing the influence of wave curvature on wave velocity. Parameters cm, ae, and k were taken from literature. Parameters am and ce were estimated by fitting the model to experimental data, obtained by pacing the canine left ventricular free wall (LVFW). The best fit was found with cm = 0.75 m/s, ce = 1.3 m/s, am = 2.5, ae = 1.5, and k = 2.1 x 10(-4) m2/s. With these parameter settings, for right ventricular apex (RVA) pacing, the depolarization times were realistically simulated as also shown by the wavefronts and the time needed to activate the LVFW. The moment of depolarization was used to initiate myofiber contraction in a model of LV mechanics. For both pacing situations, mid-wall circumferential strains and onset of myofiber shortening were obtained. CONCLUSION: With a relatively simple model setup, simulated depolarization timing patterns agreed with measurements for pacing at the LVFW and RVA in an LV. Myocardial cross-fiber wave velocity is estimated to be 0.40 times the velocity along the myofiber direction (0.75 m/s). Subendocardial wave velocity is about 1.7 times faster than in the rest of the myocardium, but about 3 times slower than as found in Purkinje fibers. Furthermore, model and experiment agreed in the following respects. (1) Ventricular pacing decreased both systolic pressure and ejection fraction relative to natural sinus rhythm. (2) In early depolarized regions, early shortening was observed in the isovolumic contraction phase; in late depolarized regions, myofibers were stretched in this phase. Maps showing timing of onset of shortening were similar to previously measured maps in which wave velocity of contraction appeared similar to that of depolarization. PMID- 14760921 TI - Study of unipolar electrogram morphology in a computer model of atrial fibrillation. AB - INTRODUCTION: Electrograms exhibit a wide variety of morphologies during atrial fibrillation (AF). The basis of these time courses, however, is not completely understood. In this study, data from computer models were studied to relate features of the signals to the underlying dynamics and tissue substrate. METHODS AND RESULTS: A computer model of entire human atria with a gross fiber architecture based on histology and membrane kinetics based on the Courtemanche et al. atrial model was developed to simulate paced activation and simulated AF. Unipolar electrograms were computed using a current source approximation at 256 sites in right atrium, to simulate a mapping array. The results show the following: (1) In a homogeneous and isotropic tissue, the presence of highly asymmetric electrograms is rare (<2%), although there is a marked variability in amplitude and symmetry. (2) The introduction of anisotropy increases this variability in symmetry and amplitude of the, electrograms especially for propagation across fibers. The percentage of highly asymmetric electrograms increases to 12% to 15% for anisotropy ratios greater than 3:1. (3) Multiphasic and fractionated electrograms are rarely seen in the model with uniform properties but are more common (15%-17%) in a model including regions with abrupt changes in conductivity. Beat-to-beat variations in the occurrence of multiphasic signals are possible with fixed anatomic heterogeneity, due to beat-to-beat variations in the direction of the wavefront relative to the heterogeneity. CONCLUSION: Analysis of the amplitude and symmetry of unipolar atrial electrograms can provide information about the electrophysiologic substrate maintaining AF. PMID- 14760925 TI - Altered T wave dynamics in a contracting cardiac model. AB - INTRODUCTION: The implications of mechanical deformation on calculated body surface potentials are investigated using a coupled biophysically based model. METHODS AND RESULTS: A cellular model of cardiac excitation-contraction is embedded in an anatomically accurate two-dimensional transverse cross-section of the cardiac ventricles and human torso. Waves of activation and contraction are induced by the application of physiologically realistic boundary conditions and solving the bidomain and finite deformation equations. Body surface potentials are calculated from these activation profiles by solving Laplace's equation in the passive surrounding tissues. The effect of cardiac deformation on electrical activity, induced by contraction, is demonstrated in both single-cell and tissue models. Action potential duration is reduced by 7 msec when the single cell model is subjected to a 10% contraction ramp applied over 400 msec. In the coupled electromechanical tissue model, the T wave of the ECG is shown to occur 18 msec earlier compared to an uncoupled excitation model. To assess the relative effects of myocardial deformation on the ECG, the activation sequence and tissue deformation are separated. The coupled and uncoupled activation sequences are mapped onto the undeforming and deforming meshes, respectively. ECGs are calculated for both mappings. CONCLUSION: Adding mechanical contraction to a mathematical model of the heart has been shown to shift the T wave on the ECG to the left. Although deformation of the myocardium resulting from contraction reduces the T wave amplitude, cell stretch producing altered cell membrane kinetics is the major component of this temporal shift. PMID- 14760924 TI - Relationship between regional shortening and asynchronous electrical activation in a three-dimensional model of ventricular electromechanics. AB - INTRODUCTION: Asynchronous electrical activation can cause abnormalities in perfusion and pump function. An electromechanical model was used to investigate the mechanical effects of altered cardiac activation sequence. METHODS AND RESULTS: We used an anatomically detailed three-dimensional computational model of the canine ventricular walls to investigate the relationship between regional electrical activation and the timing of fiber shortening during normal and ventricular paced beats. By including a simplified Purkinje fiber network and anisotropic impulse conduction in the model, computed electrical activation sequences were consistent with experimentally observed patterns. Asynchronous time courses of regional strains during beats stimulated from the left or right ventricular epicardium showed good agreement with published experimental measurements in dogs using magnetic resonance imaging tagging methods. When electrical depolarization in the model was coupled to the onset of local contractile tension development by a constant time delay of 8 msec, the mean delay from depolarization to the onset of systolic fiber shortening was 14 msec. However, the delay between the onset of fiber tension and initial shortening varied significantly; it was as late as 60 msec in some regions but was also as early as -50 msec (i.e., 42 msec before depolarization) in other regions, particularly the interventricular septum during free-wall pacing. CONCLUSION: The large variation in delay times was attributable to several factors including local anatomic variations, the location of the site relative to the activation wavefront, and regional end-diastolic strain. Therefore, we conclude that these factors, which are intrinsic to three-dimensional ventricular function, make the regional sequence of fiber shortening an unreliable surrogate for regional depolarization or electromechanical activation in the intact ventricles. PMID- 14760926 TI - Quantitative reconstruction of cardiac electromechanics in human myocardium: assembly of electrophysiologic and tension generation models. AB - INTRODUCTION: Myocytes from normal and failing myocardium show significant differences in electromechanical behavior. Mathematical modeling of the behavior provides insights into the underlying physiologic and pathophysiologic mechanisms. Electromechanical models of cardiomyocytes exist for various species, but models of human myocytes are lacking. METHODS AND RESULTS: A mathematical model of electromechanics in normal and failing cardiac myocytes in humans was created by assembly and adaptation of parameters of an electrophysiologic model at the level of single cells and a force development model at the level of the sarcomere. The adaptation was performed using data from recent studies of ventricular myocytes and myocardium. The model was applied to quantitatively reconstruct measurement data from different experimental studies of normal and failing myocardium. Several simulations were performed to quantify the transmembrane voltage Vm, intracellular concentration of calcium[Ca2+]i, the [Ca2+]i-force relationship, and force transients. Furthermore, frequency dependencies and restitution of action voltage duration to 90% recovery APD90, peak [Ca2+]i, duration to 50% force recovery FD50, and peak force were determined. CONCLUSION: The presented mathematical model was capable of quantitatively reconstructing data obtained from different studies of electrophysiology and force development in normal and failing myocardium of humans. In future work, the model can serve as a component for studying macroscopic mechanisms of excitation propagation, metabolism, and electromechanics in human myocardium. PMID- 14760927 TI - Quantitative reconstruction of cardiac electromechanics in human myocardium: regional heterogeneity. AB - INTRODUCTION: Regional heterogeneity of electrophysiologic properties within the human ventricles is based on changes in ion channel kinetics and density inside the wall. The heterogeneity not only influences the electrophysiologic properties but also cellular force development. In this study, the influence of heterogeneity was investigated using mathematical models. METHODS AND RESULTS: An overview of measurements of the heterogeneity of electrophysiology and force development is presented. This knowledge is transferred to an electromechanical heart model composed of a human ionic cell model describing electrophysiologic properties and a model for the development of forces. Heterogeneity is included in the ionic model by changing ion channel kinetics and density. The characteristics and dependencies of the electromechanical model are demonstrated in a single-cell environment and a multicell environment. In the single-cell environment, the effects of heterogeneity on electrical activity are demonstrated. The notch in the action potential decreases from epicardium to endocardium, and action potential duration is longest in the mid-myocardium. The developed forces are largest in the subendocardial cells and decrease continuously toward the epicardium. The multicell environment describes a transmural line of cells in the left ventricular free wall using a bidomain approach. The transmural ECG shows typical characteristics with a positive monophasic T wave. CONCLUSIONS: This work demonstrates the need to incorporate regional heterogeneity in order to model human cardiac electromechanics. The results of electrophysiologic simulations correspond to measured data. The dependencies of regional heterogeneity on force development need to be validated in experiments, because little is known about the influence of heterogeneity on electromechanical coupling. PMID- 14760929 TI - Effects of lidocaine on shock-induced vulnerability. AB - INTRODUCTION: Lidocaine is known to increase the defibrillation threshold (DFT) of monophasic shocks (MS) and have no effect on DFT of biphasic shocks (BS). The aim of this study was to enhance our understanding of the mechanisms of vulnerability and defibrillation through the investigation of this difference. METHODS AND RESULTS: We studied the effect of 15 microM lidocaine on shock induced vulnerability using fluorescent imaging of Langendorff-perfused rabbit hearts. Vulnerability was assessed as vulnerable window with shock strengths of 15 to 150 V and vulnerable period (VP) with shock delivery phase of 0% to 100% of action potential duration (% APD). With MS, lidocaine caused a significant increase in both the upper limit of vulnerability (ULV, 71 +/- 17 V vs 120 +/- 1.5 V, P < 0.01) and upper limit of VP (91 +/- 8.0% APD vs 110 +/- 4.2% APD, P < 0.01). With BS, lidocaine had no effect on ULV (40 +/- 3.4 V vs 45 +/- 4.5 V) and did not increase the upper limit of VP (78 +/- 8.9% APD vs 96 +/- 12% APD, P < 0.01). Lidocaine caused reduction of the conduction velocity during pacing (0.58 +/- 0.08 m/s vs 0.44 +/- 0.05 m/s, P < 0.01), shock-induced break excitation (0.82 +/- 0.17 m/s vs 0.30 +/- 0.07 m/s, P < 0.01), and postshock reentry (0.34 +/- 0.07 m/s vs 0.19 +/- 0.08 m/s, P < 0.01). Lidocaine had no effect on shock induced virtual electrode polarization. CONCLUSION: Lidocaine increased MS ULV due to slowing of shock-induced break-excitation wavefronts, which resulted in enhanced probability of survival of virtual electrode induced phase singularity. Lidocaine had no effect on BS ULV because no break excitation was induced by BS. Reduction of conduction velocity by lidocaine resulted in increased dispersion of repolarization and led to upper limit of VP increase for both MS and BS. PMID- 14760928 TI - Spontaneous initiation and termination of complex rhythms in cardiac cell culture. AB - INTRODUCTION: Complex cardiac arrhythmias often start and stop spontaneously. These poorly understood behaviors frequently are associated with pathologic modification of the structural heterogeneity and functional connectivity of the myocardium. To evaluate underlying mechanisms, we modify heterogeneity by varying the confluence of embryonic chick monolayer cultures that display complex bursting behaviors. A simple mathematical model was developed that reproduces the experimental behaviors and reveals possible generic mechanisms for bursting dynamics in heterogeneous excitable systems. METHODS AND RESULTS: Wave propagation was mapped in embryonic chick myocytes monolayers using calcium sensitive dyes. Monolayer confluence was varied by plating cultures with different cell densities and by varying times in culture. At high plating densities, waves propagate without breaks, whereas monolayers plated at low densities display spirals with frequent breaks and irregular activation fronts. Monolayers at intermediate densities display bursting rhythms in which there is paroxysmal starting and stopping of spiral waves of activity. Similar spatiotemporal patterns of activity were also observed as a function of the time in culture; irregular activity dominates the first 30 hours, followed by repetitive bursting dynamics until 54 hours, after which periodic target patterns or stable spirals prevail. In some quiescent cultures derived from older embryos, it was possible to trigger pacemaker activity following a single activation. We are able to reproduce all of these behaviors by introducing spatial heterogeneity and varying neighborhood size, equivalent to cell connectivity, in a spontaneous cellular automaton model containing a rate-dependent fatigue term. CONCLUSION: We observe transitions from irregular propagating waves, to spiral waves that spontaneously start and stop, to target waves originating from localized pacemakers in cell culture and a simple theoretical model of heterogeneous excitable media. The results show how physiologic properties of spontaneous activity, heterogeneity, and fatigue can give rise to a wide range of different complex dynamic behaviors similar to clinically observed cardiac arrhythmias. PMID- 14760931 TI - Delayed success in termination of three-dimensional reentry: role of surface polarization. AB - INTRODUCTION: Defibrillation shocks slightly stronger than cardioversion threshold may defibrillate not immediately but after a transient period of postshock activity (delayed success). The effect of a defibrillation shock is that it polarizes the tissue, primarily at the surfaces; therefore, surface polarization may play an important role at near-threshold shock intensities. METHODS AND RESULTS: We numerically investigate the effect of a monophasic transmural electrical shock on a three-dimensional (3D) reentrant wave (scroll wave). For simplicity, we assume uniform polarization of the epicardial and endocardial surfaces. We demonstrate that the effect of surface polarization alone is sufficient to induce delayed termination of self-sustained activity (3-4 beats after the shock). In agreement with experimental observations, both successful and failed shocks cause prolongation of the action potentials on the depolarized side and shortening on the hyperpolarized side, while at the same time inducing a shift from a reentrant to a focal activation pattern. Our simulations suggest that the outcome of the shock is determined by its effect on the shape of the scroll wave's center of rotation (filament). We propose a simple rule to predict the postshock filament shape that allows us to make accurate predictions of success and failure of a termination attempt. CONCLUSION: Surface polarization due to an electrical shock can terminate a reentrant scroll wave. This mechanism may explain the phenomenon of delayed success in defibrillation. PMID- 14760930 TI - Postshock arrhythmogenesis in a slice of the canine heart. AB - INTRODUCTION: Recent evidence has demonstrated that defibrillation shocks terminate or reset reentrant activity in the myocardium through the generation of virtual electrode polarization (VEP). Previous research has revealed that the shock establishes phase singularities (PSs) in the tissue via the VEP mechanism. The aim of this study was to examine, as a function of shock strength and electrode configuration, the relationship between end-shock PSs and the reentrant circuits established after failed defibrillation attempts. METHODS AND RESULTS: The study uses a complex three-dimensional finite-element bidomain model of a slice of the canine heart characterized by realistic geometry and fiber architecture and undergoing a single scroll wave. Defibrillation shocks of increasing strength are delivered through three different electrode configurations. The results demonstrated that >98% of all PSs have a lifetime of half a reentrant cycle or less. Stronger shocks result in a faster rate of annihilation of postshock PSs. For failed shocks, the surviving PSs underlie the activity of one or more scroll waves, which remain stationary in the slice. For all electrode configurations tested, the increase in shock strength leads to a rapid initial increase in the number of postshock reentries followed by a slower decrease; similar behavior is observed with regard to end-shock PSs. CONCLUSION: These results present new evidence regarding the mechanisms underlying failure of defibrillation shocks. PMID- 14760933 TI - Influence of ionic and non-ionic radiographic contrast media on leukocyte adhesion molecules. AB - BACKGROUND: Many papers have focused on the importance of granulocytes in the process of reperfusion and ischemia. Most of the clinical studies measured several parameters of this process during and after coronary angiography, without taking into account the effect of the radiographic contrast media (RCM) used during this procedure. MATERIALS AND METHODS: We performed a randomized patient study (n = 37) to evaluate the effect of ionic and non-ionic RCM on granulocyte adhesion during coronary angiography. We also evaluated the influence of the ionicity and osmolarity of the different substances on granulocyte adhesion molecules in in vitro experiments. RESULTS: The osmolarity of patient serum samples increased from 302 +/- 1 to 309 +/- 1 mOsm/kg (p < 0.05) after infusion of RCM. The CD11b expression in the samples of the non-ionic RCM treated group increased from 221 +/- 21 MFI to 377 +/- 30 MFI (p < 0.05) measured as the absolute mean fluorescence intensity (MFI), yet did not alter significantly in the ionic RCM group. In contrast, the in vitro experiments showed a reduction of the CD11b expression from 360 +/- 70 MFI to 149 +/- 30 MFI (p < 0.05) in the ionic RCM group. CONCLUSIONS: The upregulation of adhesion molecules was significantly reduced in vivo with ionic RCM, while ionic substances caused opposite effects in vitro. This effect should be taken into account when performing leukocyte functional analysis of samples taken during angiography. PMID- 14760934 TI - Regulation of nuclear factor-kappaB in intestinal epithelial cells in a cell model of inflammation. AB - BACKGROUND: Interleukin-1 (IL-1), an inflammatory cytokine whose levels are elevated in inflamed mucosa, causes part of its effect on intestinal epithelial cells (IEC) through inducing ceramide production. AIM: To study the role of nuclear factor-kappaB (NF-kappaB), a pro-inflammatory and anti-apoptotic factor, in IL-1-treated IEC. METHODS: NF-kappaB activity and levels of apoptotic proteins were assessed by electrophoretic mobility shift assay and RNA-protection assay, respectively. RESULTS: IL-1 and ceramide, which have been shown to partially mediate IL-1 effects on IEC, activated NF-kappaB levels significantly. This activation was due to a decrease in IkappaB-alpha and IkappaB-beta protein levels. Moreover, the ratio of mRNA levels of anti-apoptotic to pro-apoptotic proteins was significantly increased in IL-1-treated IEC. CONCLUSION: NF-kappaB may play a key role in the regulation of the expression of pro-inflammatory and/or apoptotic genes in inflammatory bowel disease, making this protein an attractive target for therapeutic intervention. PMID- 14760932 TI - Cytokine expression in allergic inflammation: systematic review of in vivo challenge studies. AB - BACKGROUND: Allergic inflammatory responses are driven by cells of the immune system that rely on cytokines to regulate the activity of other immune and structural cells. OBJECTIVE: To review published studies to (1) identify cytokines consistently increased after allergen challenge in atopic patients and (2) investigate temporal variation in cytokine expression. METHODS: A PUBMED systematic search was used to extract data from studies involving analysis of cytokine expression in fluids or biopsies following in vivo allergen challenge in atopic patients. RESULTS: Data were extracted from 82 studies. There were no consistent reports of cytokine protein increase in fluids of patients at 0-1 h after challenge. At 4-12 h, the chemokines eotaxin, macrophage inflammatory protein-1alpha, RANTES (regulated on activation normal T cell expressed and secreted) and interleukin (IL)-8 have all been consistently reported to be up regulated. At 18-24 h after challenge, the lymphokines IL-4, IL-5 and IL-13, as well as the pro-inflammatory cytokines granulocyte-macrophage colony-stimulating factor, tumour necrosis factor-alpha and IL-6 are consistently increased when compared with the respective control value. There were no reports of up regulation in interferon-gamma protein and mRNA and in IL-2 mRNA. CONCLUSION: The expression of granulocyte-macrophage colony-stimulating factor is consistently increased in tissues at 4-12 h after challenge. The influence of this cytokine on antigen capture and presentation by dendritic cells should be further investigated. Additionally, allergen challenge studies are needed that investigate the expression of macrophage-derived chemokine and thymus-regulated and activation-regulated chemokine in tissues of atopic patients. Blocking the effects of these lymphocyte-specific chemokines might provide new therapeutic approaches for the control of allergic inflammation. PMID- 14760935 TI - T helper cell polarisation as a measure of the maturation of the immune response. AB - BACKGROUND: T helper cell polarisation is important under chronic immune stimulatory conditions and drives the type of the evolving immune response. Mice treated with superantigens in vivo display strong effects on Th subset differentiation. The aim of the study was to detect the intrinsic capacity of T cells to polarise under various ex vivo conditions. METHODS: Purified CD4+ T cells obtained from super-antigen-treated mice were cultured under Th polarising conditions in vitro. By combining intracellular cytokine staining and subsequent flow cytometric analysis with quantitative cytokine measurements in culture supernatants by enzyme-linked immunosorbent assay (ELISA), the differential Th polarising capacity of the treatment can be detected in a qualitative and quantitative manner. RESULTS AND CONCLUSIONS: BALB/c mice were shown to be biased to develop strong Th2 polarised immune responses using Th0 stimulation of purified CD4+ T cells from phosphate-buffered saline-treated mice. Nevertheless, our analysis methodology convincingly showed that even in these mice, Toxic Shock Syndrome Toxin-1 treatment in vivo resulted in a significantly stronger Th1 polarising effect than control treatment. Our results indicate that populations of Th cells can be assessed individually for their differential Th1 or Th2 maturation capacity in vivo by analysing robust in vitro polarisation cultures combined with intracellular cytokine staining and ELISA. PMID- 14760937 TI - Histamine mediates the pro-inflammatory effect of latex of Calotropis procera in rats. AB - INTRODUCTION: Calotropis procera is known to produce contact dermatitis and the latex of this plant produces intense inflammation when injected locally. However, the precise mode of its pro-inflammatory effect is not known. In present study we have pharmacologically characterized the inflammation induced by latex of C. procera in a rat paw edema model and determined the role of histamine in latex induced inflammation. METHODS: Inflammation was induced in the hind paw of rats by injecting different doses of dried latex (DL) of C. procera. The inhibitory effect of phenylbutazone, dexamethasone, celecoxib, cyproheptadine, chlorpheniramine and compound 48/80 on edema volume was evaluated and compared with that against carrageenan. The histamine content of DL was measured fluorometrically. RESULTS: DL produced dose-dependent inflammation of the rat paw. Cyproheptadine and chlorpheniramine effectively inhibited DL-induced inflammation (90%; p < 0.01), while anti-inflammatory drugs phenylbutazone, dexamethasone and celecoxib were more effective against carrageenan-induced inflammation. Depletion of mast cell histamine by compound 48/80 produced a significant decrease in DL-induced inflammation as compared with carrageenan (500% versus 25%). DL was also found to contain about 6 microg/g of histamine. CONCLUSIONS: Thus, our study shows that the biogenic amines play a significant role in C. procera latex-induced inflammation and antihistaminic drugs could be effectively used to inhibit inflammatory response elicited by exposure to latex. PMID- 14760936 TI - Vascular endothelial growth factor and its soluble receptors VEGFR-1 and VEGFR-2 in the serum of patients with systemic lupus erythematosus. AB - We investigated the serum concentration of vascular endothelial growth factor (VEGF) and its two soluble receptors, sVEGFR-1 and sVEGFR-2, in a group of 60 patients with systemic lupus erythematosus (SLE), and 20 healthy controls, using an enzyme-linked immunosorbent assay. We examined a possible association between serum levels of these proteins and certain clinical and laboratory parameters as well as SLE activity. VEGF, sVEGFR-1 and sVEGFR-2 were detectable in all patients with SLE and in all normal individuals. The VEGF level was higher in active SLE (mean, 300.8 pg/ml) than in inactive SLE (mean, 165.9 pg/ml) (p < 0.05) or in the control group (mean, 124.7 pg/ml) (p < 0.04). The highest sVEGFR-1 concentrations were also detected in active SLE patients (mean, 42.2 pg/ml) and the lowest in inactive disease (mean, 32.0 pg/ml) (p < 0.01). In contrast, the levels of sVEGFR 2 were lower in SLE (mean, 12557.6 pg/ml) than in the control group (mean, 15025.3 pg/ml) (p < 0.05). We found a positive correlation between sVEGFR-1 concentration and the SLE activity score p = 0.375 (p < 0.004) and a negative, but statistically insignificant correlation between sVEGFR-2 and SLE activity (p = -0.190, p > 0.05). Treatment with steroids and cytotoxic agents did not influence VEGF or its soluble receptors levels. In conclusion, in SLE patients the levels of VEGF and sVEGFR-1 are higher in patients with active SLE than in inactive disease or healthy persons. In contrast, the level of sVEGFR-2 is lower in active SLE than in inactive disease. The imbalance between VEGF and its soluble receptors may be important in SLE pathogenesis. PMID- 14760938 TI - Tumour necrosis factor-alpha polymorphism as one of the complex inherited factors in pemphigus. AB - The aim of our study was to analyse a significance of tumour necrosis factor (TNF)-alpha promoter gene polymorphisms in relation to the HLA-DR locus in genetic predisposition to pemphigus. TNF-alpha gene polymorphisms in position 238 and -308 were identified using a modified polymerase chain reaction restriction fragment length polymorphism method in 53 patients with pemphigus (38 with pemphigus vulgaris, 15 with pemphigus foliaceus) and 87 healthy controls. The HLA-DRB1 locus was typed using the polymerase chain reaction SSO method in all the patients and 152 population controls. Carriers of the TNF-alpha polymorphic -308 A allele were found to be more frequent in the pemphigus foliaceus group in comparison with the control group (odds ratio (OR) = 8.12; p = 0.0005). A significant association between HLA-DRB1*04 (OR = 3.86; pcor = 0.0001) and DRB1*14 (OR = 8.4; pcor = 0.0001) and pemphigus vulgaris was found. In this group of patients a decreased frequency of HLA-DRB1*07 (OR = 0.08; pcor = 0.006) was also identified. We have shown for the first time a positive association of TNF-alpha polymorphism in position -308 with pemphigus foliaceus. PMID- 14760939 TI - Simultaneous measurement of multiple Th1 and Th2 serum cytokines in psoriasis and correlation with disease severity. AB - BACKGROUND: Psoriatic plaques have been shown to contain increased levels of proinflammatory cytokines. Serum levels of interleukin (IL)-6, IL-7, IL-8, and interferon (IFN)-gamma have been reported elevated in psoriatic patients. AIM: To evaluate serum cytokine profiles in psoriasis patients by improved enzyme-linked immunosorbent assay (ELISA) technique and to correlate these levels with disease severity. METHODS: We analyzed single serum samples from 10 patients with active untreated psoriasis, two patients with active treated psoriasis, and five healthy volunteers for major T helper type 1 and T helper type 2 cytokines using the LINCOplex ELISA multi-analyte detection system that permits simultaneous detection of multiple cytokines from a single sample. The disease severity, including erythema, induration, scale, and surface area, was assessed. RESULTS: IFN-gamma was markedly elevated in all sera from psoriasis patients, 33.8 +/- 1.3 pg/ml (mean +/- standard error) versus 8 +/- 1.5 pg/ml for normal controls (p < 0.01), and positively correlated with all indices of disease severity (Spearman r > 0.6). IL-8 was also increased in psoriasis patients (24.4 +/- 1.8 pg/ml) versus normal controls (3.6 +/- 1.2 pg/ml) (p < 0.05) and positively correlated with the degree of erythema (Spearman r > 0.6). Mean IL-12 levels were decreased in sera from psoriasis patients (8.5 +/- 1.2 pg/ml) compared with normal controls (42.2 +/- 5.3 pg/ml) (p < 0.01). Also, serum IL-10 levels were below detection levels in psoriatics compared with controls (6.4 +/- 1.3 pg/ml). CONCLUSIONS: This new ELISA system allowed rapid and reliable detection of numerous cytokines in single serum samples from patients with psoriasis. We observed that IFN-gamma and IL-8 cytokines were elevated in psoriatics and correlated with parameters of disease severity while IL-10 and IL-12 were decreased. PMID- 14760940 TI - Gram-negative, but not Gram-positive, bacteria elicit strong PGE2 production in human monocytes. AB - Gram-positive and Gram-negative bacteria induce different cytokine patterns in human mononuclear cells. We have seen that Gram-positives preferentially induce IL-12 and TNF-alpha, whereas Gram-negatives induce more IL-10, IL-6, and IL-8. In this study, we compared the capacity of these two groups of bacteria to induce PGE2. Monocytes stimulated with Gram-negative bacterial species induced much more PGE2 than did Gram-positive bacteria (5600 +/- 330 vs. 1700 +/- 670 pg/mL, p < 0.001). Blocking of COX-2 by NS398 abolished PGE2 production, but did not alter the cytokine patterns induced by Gram-positive and Gram-negative bacteria. We suggest that Gram-positive and Gram-negative bacteria may stimulate different innate effector functions; Gram-positive bacteria promoting cell-mediated effector functions whereas Gram-negative bacteria inducing mediators inhibiting the same. PMID- 14760941 TI - Modulation of macrophage activity by proteolytic enzymes. Differential regulation of IL-6 and reactive oxygen intermediates (ROIs) synthesis as a possible homeostatic mechanism in the control of inflammation. AB - Inflammatory foci are rich in proteases released by neutrophils (serine proteases) and macrophages (metalloproteases). These enzymes can degrade extracellular matrix proteins and cell membrane bound proteins thus contributing to the development and progression of inflammatory reaction. In this study we have investigated the influence of collagenase (metalloprotease) and trypsin (serine protease) on murine resident and oil-induced peritoneal macrophages (Mf). Short in vitro treatment of Mf, not affecting cell viability, significantly reduced the release of reactive oxygen intermediates (ROIs) and at the same time triggered the increase of IL-6 production and to lesser extent of TNF-alpha production. Both these effects were dependent on enzyme concentration used and were particularly well pronounced in resident macrophages. In addition both enzymes cleaved a number of cell-membrane molecules, including CD23, CD14, CD95L, and Mac-3. We hypothesize that the enzymatic digestion of certain Mf surface receptor proteins in inflammatory foci may be responsible for modification of cell behaviour either by preventing the generation of specific signal or alternatively by delivering a mock substitute signal to the cell interior. In effect inhibition of ROIs production limits their destructive effects and the increase in the secretion of IL-6 stimulates the synthesis of acute phase proteins and triggers other anti-inflammatory mechanisms thus directing Mf present in inflammatory foci into regulatory pathway rather than allowing them to perform solely the effector function. PMID- 14760942 TI - Syntheses of prostaglandin E2 and E-cadherin and gene expression of beta-defensin 2 by human gingival epithelial cells in response to Actinobacillus actinomycetemcomitans. AB - The interaction between epithelial cells and microorganisms is the most important step in bacterial infections. Actinobacillus actinomycetemcomitans was suggested to play a significant role in the initiation of periodontitis because of its bacteriological characteristics. Prostaglandins (PG) mediate the inflammatory response. Human beta-defensin-2 (hBD-2) is an antimicrobial peptide and contributes to innate immunity. E-cadherin is responsible for an epithelial intercellular junction. In this study, we investigated the syntheses of PGE2 and E-cadherin and the expression of hBD-2 in human gingival epithelial cells (HGEC) following exposure to A. actinomycetemcomitans. The levels of PGE2 and cyclooxygenase-2, which are responsible for an increase in PGE2, were increased depending on bacteria exposure time. hBD-2 mRNA was induced by A. actinomycetemcomitans, while HGEC exposed to A. actinomycetemcomitans showed a decrease in E-cadherin levels. Etodolac, a selective cyclooxygenase-2 inhibitor reinforced the increase in hBD-2 mRNA levels by A. actinomycetemcomitans. Furthermore, the etodolac suppressed the decrease in E-cadherin levels. Thus, endogenous PGE2 is involved in the hBD-2 and E-cadherin responses of HGEC to A. actinomycetemcomitans. These findings suggest that the inflammatory and antimicrobial response of gingival epithelial cells to A. actinomycetemcomitans is involved in the initiation of periodontal inflammation. A. actinomycetemcomitans may destroy the mechanical epithelial barrier by destroying E-cadherin. PMID- 14760945 TI - Oligoclonal bands in Devic's neuromyelitis optica and multiple sclerosis: differences in repeated cerebrospinal fluid examinations. AB - We studied repeated cerebrospinal fluids of patients with Devic's neuromyelitis optica (NMO) and multiple sclerosis (MS). Variations of oligoclonal bands (OBs) had opposite trends in the two groups. In MS, OBs were detected in 399 of 411 patients (97%) and never disappeared. In NMO, OBs were detected in three of 11 patients (27%) and always disappeared. The hypothesis that NMO and MS follow distinct pathogenetic pathways is supported by our findings, which can be useful for the differentiation of NMO from MS. PMID- 14760943 TI - Estradiol treatment ameliorates acetic acid-induced gastric and colonic injuries in rats. AB - To evaluate the role of estrogen treatment on the healing of acetic acid-induced gastric or colonic injury, rats were given 17beta estradiol benzoate (0.001, 0.1, and 10 mg/kg) or vehicle for 7 days (following the induction of ulcer) or 4 days (following the induction of colitis) until they were decapitated. Food intake and fecal output were decreased by estradiol treatment but gastric emptying rate was not changed. Estradiol (10 mg/kg) reduced gastric ulcer index and colonic damage score compared to vehicle-treated groups. SEM and light microscopy demonstrated a significant reduction in the severity of ulcers and colitis by estradiol treatment. Gastric microscopic score was not changed by estradiol treatment, whereas in the colonic tissue score was significantly reduced. Elevated gastric MPO levels were reduced in gastric but not in colonic tissues as compared with corresponding vehicle groups. In conclusion, exogenous estradiol treatment at pharmacological doses improves the healing of both gastric and colonic injury induced by acetic acid in rats. PMID- 14760944 TI - Identification and subcellular localization of neuronal calcium sensor-1 (NCS-1) in human neutrophils and HL-60 cells. AB - Secretion in neutrophils is thought to be regulated in different ways for the different granule types. Specific granules are endowed with proteins which are related to docking and fusion events and are absent on azurophilic granules. Furthermore, even if secretion of content from all neutrophil granules is a Ca(2+)-dependent process, a higher concentration of cytosolic calcium is required for azurophilic than for specific granule secretion. In this paper we show that human neutrophils and promyelocitic cells express neuronal calcium sensor-1 (NCS 1), a calcium binding protein involved in exocytosis in various cell types. Both mRNA and protein were found in mature cells and precursors. NCS-1 is shown to be mainly associated with azurophilic granules and, therefore could play an instrumental role in the calcium-dependent secretion of azurophilic granules. PMID- 14760946 TI - Concordance for disease course and age of onset in Scandinavian multiple sclerosis coaffected sib pairs. AB - BACKGROUND: Investigation of coaffected sib pairs is one method to determine the genetic influence on the clinical presentation of many complex diseases, such as multiple sclerosis (MS). Investigation of the clinical concordance in coaffected sib pairs may be a prerequisite to identify genes that modify the clinical outcome. The aim of this study was to investigate a possible genetic influence on selected demographic and clinical variables among familial Scandinavian MS cases. MATERIAL AND METHODS: We identified 136 Caucasian Scandinavian families with MS coaffected sib pairs from Denmark, Norway and Sweden. Cohen's kappa coefficient and the intraclass correlation coefficient were used to assess concordances in sib pairs. Furthermore, clinical features and HLA-DR2 carrier status were compared among the probands of sib pairs. RESULTS: We found significant concordance of the disease course (kappa = 0.28, P < 0.001) and adjusted age of onset (r = 0.23, P = 0.028). Among probands of sib pairs, HLA-DR2 carrier patients had a younger age of onset (P = 0.024). CONCLUSION: Analyses of Scandinavian coaffected sib pairs suggest that disease course and age of onset are partly under genetic control. Furthermore, HLA-DR2 in probands of sib pairs suggests importance for age of onset. PMID- 14760947 TI - Diffusion tensor imaging of early relapsing-remitting multiple sclerosis with histogram analysis using automated segmentation and brain volume correction. AB - Diffusion tensor magnetic resonance imaging (DTI) reveals measurable abnormalities in normal-appearing brain tissue (NABT) in established multiple sclerosis (MS). However, it is unclear how early this occurs. Recent studies have employed whole brain histogram analysis to improve sensitivity, but concern exists regarding reliability of tissue/cerebrospinal fluid segmentation and possible intersubject brain volume differences, which can introduce partial volume error: To address this, 28 early relapsing-remitting MS subjects [median disease duration 1.6 years; median Expanded Disability Status Scale (EDSS) score 1.5] and 20 controls were compared with whole brain histogram analysis using an automated segmentation algorithm to improve reproducibility. Brain parenchymal volumes (BPV) were estimated for each subject in the analysis. The mean, peak height and peak location were calculated for DTI parameters [fractional anisotropy (FA), mean diffusivity and volume ratio]. An increased FA peak height in MS subject NABT was observed (P = 0.02) accounting for age, gender and BPV. Removing BPV revealed additional abnormalities in NABT. The main conclusions are i) FA peak height is increased in NABT in early MS, ii) partial volume edge effects may contribute to apparent NABT histogram abnormalities, and iii) correction for brain volume differences should reduce potential partial volume edge effects. PMID- 14760948 TI - Multiple sclerosis: expression of CD1a and production of IL-12p70 and IFN-gamma by blood mononuclear cells in patients on combination therapy with IFN-beta and glatiramer acetate compared to monotherapy with IFN-beta. AB - Current therapy of multiple sclerosis (MS) with interferon-beta (IFN-beta) or glatiramer acetate (GA) has modest effects on the course of MS. Both compounds affect several immune variables, like expression of cell surface molecules and cytokine levels. Here we compared untreated MS, therapy with IFN-beta alone and combined with GA, and healthy controls (HC), regarding expression on HLA-DR+ blood mononuclear cells (MNC) of CD1a that is a cell surface molecule with capacity to present glycolipids to T cells, and of CD80 and CD86 which are costimulatory molecules that activate Th1 and Th2 responses. Cytokine production by MNC was also measured. Flow cytometry and ELISA were used. Cross-sectional comparisons revealed that untreated MS patients had higher CD1a+ HLA-DR+ MNC and lower IL-10 production compared to patients treated with IFN-beta or IFN-beta+GA or HC. Untreated MS patients also had higher spontaneous IFN-gamma and IL-12p70 production compared to MS patients treated with IFN-beta+GA or HC, but not when compared to MS patients on monotherapy with IFN-beta. Low CD1a+ HLA-DR+ MNC and low spontaneous production of IL-12p70 and IFN-gamma were more pronounced in patients treated with IFN-beta+GA than with IFN-beta alone. In order to clarify whether these changes reflect disease activity or treatment effects, we performed a follow up study. Nineteen MS patients with disease progression, despite monotherapy with IFN-beta for more than one year; were re-examined after one to three and four to six months of treatment with IFN-beta+GA. This combination therapy was associated with normalization of CD1a+ HLA-DR+ MNC, IL-12p70 and IFN gamma. It remains to be shown whether these immunological changes imply a clinical benefit. Follow up studies of immune variables versus clinical effects during combined therapy of MS with IFN-beta+GA are ongoing. PMID- 14760949 TI - Distinct roles for IP-10/CXCL10 in three animal models, Theiler's virus infection, EAE, and MHV infection, for multiple sclerosis: implication of differing roles for IP-10. AB - Theiler's murine encephalomyelitis virus (TMEV) causes demyelination with inflammation of the central nervous system (CNS) in mice and is used as an animal model for multiple sclerosis (MS). Interferon-gamma inducible protein-10 kDa (IP 10) is a CXC chemokine and a chemoattractant for CXCR3+ T cells. IP-10 mRNA is expressed in the CNS during TMEV infection. However, administration of anti-IP-10 serum caused no difference in clinical signs, inflammation, demyelination, virus persistence or anti-virus antibody response in TMEV infection, while levels of virus specific and autoreactive lymphoproliferation increased. This likely reflects a difference in the pathogenesis of TMEV infection from that of two other animal models for MS, mouse hepatitis virus infection and experimental allergic encephalomyelitis (EAE), where blocking of IP-10 resulted in clinical and histological improvement with suppression of antigen specific lymphoproliferation. In this review, we compare and contrast the roles of IP-10 between the three animal models for MS, and discuss the relevance to MS patients with different clinical courses. PMID- 14760950 TI - Alcohol and drug abuse among persons with multiple sclerosis. AB - OBJECTIVE: To examine the one-month prevalence and impact of substance abuse in a large community sample of persons with multiple sclerosis (MS). METHOD: Members of the Multiple Sclerosis Society of King County were surveyed by mail. This multifaceted health survey included questions pertaining to substance abuse. Seven hundred and thirty-nine out of 1374 potential participants (54%) returned the survey, while 708 reported a medically confirmed diagnosis of MS and provided sufficient data. RESULTS: Fourteen per cent of the sample screened positive for possible alcohol abuse or dependence, and 7.4% reported misusing illicit drugs or prescription medications within the previous month. Possible alcohol abuse and drug misuse were associated with younger age, less severe MS related disability and being employed, as well as greater self-reported depressive symptomatology. Most persons with alcohol problems indicated interest in learning more about ways to stop or cut down. CONCLUSIONS: Substance abuse may be present in up to 19% of this sample and contribute to high rates of depression. There may be greater risk of harm due to substance abuse in people with MS because of the potential magnification of motor and cognitive impairments. Comprehensive MS care should include substance abuse screening and advice to cut down or abstain. PMID- 14760951 TI - Outcome measures for multiple sclerosis clinical trials: relative measurement precision of the Expanded Disability Status Scale and Multiple Sclerosis Functional Composite. AB - We compared the relative measurement precision (RMP) of the Expanded Disability Status Scale (EDSS) and the Multiple Sclerosis Functional Composite (MSFC) for discriminating between groups of patients known to differ in their extent of multiple sclerosis (MS). A total of 133 patients were rated with the EDSS and MSFC and had magnetic resonance imaging (MRI) scans. Patients were grouped on the basis of MRI appearances (T1- and T2-weighted lesion loads, parenchymal and ventricular fractions--T1LL, T2LL, PF, VF, respectively) and RMP was determined using the method of group differences. For each MRI parameter, the total sample was arranged in ascending order of magnitude and divided into two, three, four and five similar sized groups. For each division (two, three, four or five groups), EDSS and MSFC scores for the groups were compared using parametric (paired samples t-tests, one-way ANOVA) and nonparametric (Wilcoxon's rank-sum test, Kruskal-Wallis analysis of variance) statistical methods and RMP was estimated. The EDSS and MSFC were correlated substantially (r = -0.64). Relative to the MSFC, the EDSS had inferior measurement precision regardless of the number of groups into which the total sample was divided, or the statistical method. However, the RMP of the EDSS compared with the MSFC varied from 2% to 86%. Results suggest the MSCF is better than the EDSS for detecting differences between groups of patients, defined by these MRI markers of MS. However, the finding that both scales correlated weakly with MRI markers, indicated that they are limited as predictors of MS pathology as defined by MRI. An explanation for this well-established clinical-MRI paradox is that rating scales and MRI measure fundamentally different manifestations of MS. PMID- 14760952 TI - A systematic approach to analyse health-related quality of life in multiple sclerosis: the GEDMA study. AB - OBJECTIVE: To describe a holistic and comprehensive approach to the assessment of sufferer's perceptions of health-related quality of life (HRQoL) in a cohort of multiple sclerosis (MS) patients. METHODS: The GEDMA (Grupo de Enfermedades Desmielinizantes de Madrid, in Spanish) study is an ongoing longitudinal survey using quantitative and qualitative methodologies. The baseline cohort consisted of a large sample of MS patients recruited from 13 hospitals in Madrid, Spain. Using a standardized protocol we collected data concerning the sociodemographic and health status characteristics of patients, as well as implementing a modified Spanish version of the Functional Assessment of Multiple Sclerosis quality of life instrument Primary caregivers were interviewed using a specific protocol combined with the Zarit Burden Interview. RESULTS: The index cohort comprised 371 MS patients (68.7% female) of mean age 38.9+/-0.9 years. Age, sex and clinical form distribution were similar to other MS population-based surveys. There were 258 (69.5%) relapsing-remitting (RR) MS patients and 113 (30.5%) progressive MS patients. More than one-third of the married patients with progressive MS and almost a quarter of the RRMS patients separated or divorced following a diagnosis of MS; 71.3% of the progressive MS patients as well as 65.8% of the RRMS patients were unemployed as a consequence of the disease. Qualitative analysis showed that friendship and family relationships and occupational status were the most significant dimensions influenced by MS. On the other hand, the speech analysis of primary caregivers showed that emotional burden was related to patients' physical disability. Furthermore, primary caregivers described the influence of MS on their own occupational status, their nonacceptance of the disease, a perception of a lack of support by other members of the family as well as a 'selfish and intransigent' attitude of the patients themselves. CONCLUSIONS: The analysis of the GEDMA cohort provides valuable information that helps clarify the impact of MS on patients' HRQoL. PMID- 14760953 TI - The patient's perception of a (reliable) change in the Multiple Sclerosis Functional Composite. AB - OBJECTIVE: To prospectively characterize the relation between two-year changes in functional impairment as measured by the Multiple Sclerosis Functional Composite (MSFC) and changes in patient perceived disability as measured by the Guy's Neurological Disability Scale (GNDS). METHODS: One hundred and eighty-eight patients with multiple sclerosis (MS) were recruited at our outpatient clinic. Impairment and disability were assessed using the MSFC and GNDS at baseline and follow-up. Longitudinal correlations were studied between changes in MSFC and GNDS and their corresponding components. We also studied changes in GNDS in relation to what can be classified as a reliable change in MSFC; for example, 20% change in each MSFC component or a change of 0.5 in total MSFC score. In addition, we studied the change in total number of GNDS subcategories with a score of 3 or higher in relation to the predefined MSFC changes, these subcategories being indicative of the requirement for help by another person. RESULTS: Despite good cross-sectional correlations between MSFC and GNDS, no significant correlation was found between longitudinal changes in MSFC and GNDS. Analysing the change in GNDS in relation to the predefined MSFC changes shows that GNDS changes are nicely rank ordered when more stringent definitions of reliable change were applied. In addition, analysing the number of GNDS subcategories scored 3 or higher indicate that there is a profile of worsening on the MSFC being associated with increase in the amount of help required from others. CONCLUSION: Our longitudinal data suggest that a reliable change is associated with a likewise change in patient perceived disability, the smallest reliable change being identified by at least 20% change in each MSFC component. PMID- 14760954 TI - Self-efficacy predicts self-reported health status in multiple sclerosis. AB - Self-efficacy is a belief that one can competently cope with a challenging situation. If self-efficacy is a strong predictor of health status in multiple sclerosis (MS), it may be an important area to target in clinical practice, as such beliefs may be modifiable. The aim of this study was to examine the predictive value of self-efficacy on self-reported health status in MS. Eighty nine people with MS completed the Multiple Sclerosis Self-efficacy Scale (MSSE function and control scales), the Multiple Sclerosis Impact Scale (MSIS-29), and the Multiple Sclerosis Walking Scale (MSWS-12) at two time points: 1) admission to an inpatient rehabilitation unit (n = 43) or for steroid treatment for relapses (n = 46); and 2) discharge (rehabilitation group) or six weeks later (steroid group). Multiple regression analyses examined whether baseline and changes in self-efficacy predict changes in self-reported health status. Both baseline and changes in self-efficacy were strong and independent predictors of changes in health status (P-values ranged from 0.025 to < 0.001). That is, pretreatment self-efficacy scores and increases in self-efficacy scores from baseline to follow-up (improvement), were significantly associated with decreases (improvement) in perceived walking ability and physical and psychological impact of MS. The findings suggest that self-efficacy predicts improvement in health status and that self-efficacy would be an important domain to measure and manage actively in education and rehabilitation programs. PMID- 14760955 TI - Cognitive strategies application of multiple sclerosis patients. AB - When performing a novel task, people need to generate and apply a working strategy. The application of an appropriate working strategy enables patients with cognitive impairment (CI) to perform tasks efficiently, which in turn makes it easier to meet the challenges of daily life tasks. We investigated the strategy application abilities (SAA) of multiple sclerosis (MS) patients and compared these results with data based on healthy subjects' performance. Seventy six patients performed a Strategy Application Test (SAT) along with other cognitive tests, and completed depression, fatigue, and activity of daily living (ADL) questionnaires. Our results indicated that 76% of the MS patients included had impaired SAA, and that this impairment was not correlated with their depression, fatigue, Expanded Disability Severity Scale (EDSS), or ADL. These findings may have important implications for the understanding of the capability of MS patients to cope with nonroutine tasks, as well as for the potential of future implementation of cognitive rehabilitation in improving the SAA of patients with MS or other cognitive disorders. PMID- 14760956 TI - Analyses of nursing home residents with multiple sclerosis at admission and one year after admission. AB - This research compares profiles of residents with multiple sclerosis (MS) at admission to the nursing facility with profiles of these same residents one year later using the Minimum Data Set (MDS) to determine how their health and care received changed after one year in the facility. We matched MDS admission assessments with their first annual assessment for 1309 residents with MS. These residents with MS demonstrated deterioration in cognitive performance, communication skills, motor performance and bladder/bowel continence after one year in the facility. However, the proportion of residents with urinary tract infections and the proportion of residents with pressure ulcers declined after one year. A significantly larger proportion of these MS residents had a diagnosis of depression one year after admission, with significantly increased use of antipsychotic and antidepressant medications but little and declining use of psychological therapy after one year in the facility. In addition, there was a significant decline after one year in the use of physical, occupational, and speech therapies. These longer stay residents with MS may benefit from support services directed toward mental health and increased use of physical, occupational and psychological therapies provided in the nursing facility. PMID- 14760957 TI - Peer support groups in multiple sclerosis: current effectiveness and future directions. AB - Peer support programs have become a common method of providing support for patients with chronic illness. Utilizing peers as resources has been proposed as an effective means for coping with a stressful life experience and for gaining support from others who share a common factor, although data are somewhat mixed on the efficacy of peer support. The aim of the present study was to evaluate the effectiveness of eight weeks of a standard form of peer support in improving quality of life and reducing depressive symptoms in 44 patients with multiple sclerosis (MS). One person from each of six groups participated in a training course in order to learn basic principles of peer support. Eight weekly sessions were held and patients completed self-administered questionnaires pre- and post treatment assessing quality of life and depression. Results showed that support groups do not provide consistent improvement in quality of life or depression in patients with MS and suggest that patients who have better mental health functioning could be at risk for deterioration in support groups. PMID- 14760958 TI - Scleromyxedema in a patient with multiple sclerosis and monoclonal gammopathy on interferon beta-1a. AB - BACKGROUND: Animal studies have shown that some human monoclonal antibodies promote myelin repair in models of demyelinating disease. Scleromyxedema is a dermatologic disorder associated with a monoclonal gammopathy and neurologic manifestations. The reason for occurrence of cutaneous reactions in interferon treated patients is unknown. CASE DESCRIPTION: A 37-year-old woman was started on weekly interferon beta-1a (IFN beta-1a) following a diagnosis of multiple sclerosis (MS). After having been on interferon therapy for three years, she developed skin lesions secondary to scleromyxedema. Her IFN beta-1a was discontinued and intravenous immunoglobulin therapy was started for her scleromyxedema. At a six-month follow up, her skin lesions improved and there was no recurrence of neurologic symptoms. CONCLUSIONS: This is the first report of occurrence of scleromyxedema in a patient with MS. While this could be a chance association, it does raise the question if her neurologic manifestations could be secondary to scleromyxedema. Further research into the mechanism of IFN related cutaneous side effects is needed. Evidence regarding the remyelinating nature of human monoclonal antibodies raises interest in the potential therapeutic role these antibodies may have. PMID- 14760959 TI - No evidence for genetic linkage between development of multiple sclerosis and components of the IFN system and the JAK-STAT pathway. AB - Multiple sclerosis (MS) is an inflammatory, demyelinating disease of the central nervous system (CNS). Several observations suggest that the interferon system may be of interest in the study of MS development To investigate whether polymorphism in components of the IFN system and the JAK-STAT pathway influence susceptibility to MS, we performed a linkage analysis between polymorphic loci in or close to the IFN gamma, IFN gamma receptor, IFN alpha/beta receptor, JAK 1, STAT 1 and STAT 3 genes in 27 Swedish families with at least two members having MS. Tests for transmission disequilibrium and nonparametric linkage analysis gave negative results. We found no evidence for linkage between MS and any of these loci. PMID- 14760960 TI - A randomized, double-blind, placebo-controlled pilot study of i.v. immune globulins in combination with i.v. methylprednisolone in the treatment of relapses in patients with MS. AB - BACKGROUND: Some patients with multiple sclerosis (MS) do not show a clear improvement of acute relapses after treatment with intravenous methylprednisolone (IVMP). We compared the efficacy of the combination of intravenous immunoglobulins (IVIg) and IVMP with the standard treatment of IVMP alone in promoting recovery from moderate to severe acute relapses in MS. METHODS: Patients with clinically definite MS having a relapse with at least a one point increase in Kurtzke's expanded disability status scale (EDSS) in comparison to the preattack EDSS were randomized to IVMP-IVIg or IVMP-placebo treatment. The primary outcome criterion was the EDSS grade at four weeks. A preplanned interim analysis was performed after inclusion of 19 consecutive MS patients to evaluate the sample size necessary for a larger trial. FINDINGS: Both groups had improved one point on the EDSS four weeks after start of treatment (P = 0.81) and one of the stopping rules of the interim analysis was fulfilled. There were also no differences in secondary outcomes: EDSS at eight and 12 weeks, time to improve > or = 1 EDSS points, difference in Scripps score and ambulation index. Five patients in the IVMP-IVIg group and two in the IVMP group had a new relapse in the six month follow-up. INTERPRETATION: Our study could not show superiority of IVMP-IVIg in the treatment of moderate to severe acute relapses in MS. PMID- 14760961 TI - Therapy related acute myeloblastic leukaemia after mitoxantrone treatment in a patient with multiple sclerosis. PMID- 14760962 TI - The Chinese in Canada: a study in ethnic change with emphasis on gender roles. AB - The authors investigated the impact of ethnic change experienced by Chinese Canadian couples on gender-role attitude, household task-role expectations and performance. The authors presented acculturation and Chinese ethnic identification as the two discriminant facets of ethnic change. Results indicated a nonsignificant role of acculturation in bringing about modifications of the gender-role attitudes of husbands and of their household task-role expectations. In contrast, the acculturation of Chinese Canadian wives proved to be a significant factor in promoting more modern (less traditional) gender-role attitudes, which in turn led to role expectations that they should contribute less to the performance of the tasks that traditionally fall in the female domain whereas their husbands should contribute more. Subsequent results also revealed that the acculturation of wives was directly linked to the role expectation that they should assume a greater share of responsibility in taking care of the traditionally husband-responsible tasks whereas their husbands should contribute a smaller share. Moreover, Chinese ethnic identification emerged as a significant determinant of husbands' gender-role attitudes and influenced their role expectation that husbands should contribute more to the performance of the tasks that traditionally fall in the male domain whereas their wives should contribute less. PMID- 14760964 TI - Gender and culture differences in touching behavior. AB - The authors used gender and culture to examine the theory that touching behavior is an expression of dominance. Participants were 120 men and women from Italy, the Czech Republic, and the United States. The authors examined both hand touches and nonhand touches. For hand touches, there was a significant gender-by-culture interaction in that Czech men as a group touched more than any of the other groups. For nonhand touches, Czech and Italian women and Italian men as groups touched significantly more than any of the other groups. Taken in cultural context, these results seem to support the dominance theory for touches with the hand but not for nonhand touches. The authors discussed implications and future directions. PMID- 14760963 TI - Stereotype accuracy of ballet and modern dancers. AB - The authors recorded preprofessional ballet and modern dancers' perceptions of the personality traits of each type of dancer and self-reports of their own standing, to test the accuracy of the group stereotypes. Participants accurately stereotyped ballet dancers as scoring higher than modern dancers on Fear of Negative Evaluation and Personal Need for Structure and accurately viewed the groups as equal on Fitness Esteem. Participants inaccurately stereotyped ballet dancers as lower on Body Esteem; the groups actually scored the same. Sensitivity correlations across traits indicated that dancers were accurate about the relative magnitudes of trait differences in the two types of dancers. A group of nondancers reported stereotypes that were usually in the right direction although of inaccurate magnitude, and nondancers were sensitive to the relative sizes of group differences across traits. PMID- 14760965 TI - The effect of accent evaluation and evidence on a suspect's perceived guilt and criminality. AB - The authors investigated the effect of accent evaluation, evidence, and crime type on participants' perceptions of guilt and criminality. British student raters (n = 199) listened to a tape-recorded exchange between a male criminal suspect and a male policeman. The authors manipulated this exchange to produce a 2 (accent type: English regional or standard) x 2 (evidence type: strong or weak) x 2 (crime type: blue collar or white collar) factorial design. Dependent measures consisted of participants' ratings of the suspect's guilt and criminality. Contrary to previous research, accent did not significantly influence attributions of guilt either as a main effect or in interaction with the contextual variables. However, independent of evidence presented and crime type, the regional-accented suspect was evaluated as more typically criminal and more likely to be reaccused of a crime than the standard-accented suspect. PMID- 14760966 TI - Effects of defendant age on severity of punishment for different crimes. AB - After reading a murder or theft vignette in which the perpetrator was a 20-, 40-, or 60-year-old man, 95 undergraduates gave sentence and parole recommendations. Punishment was harsher for the murder than for the theft. For murder, participants treated the 20- and 60-year-old men less harshly than the 40-year old man, which confirms previous archival findings. However, this inverted U shaped function occurred for murder only. The authors discussed the results in the context of the just-desert and utilitarian rationales that guide sentencing and gave suggestions for future research. PMID- 14760967 TI - Effect of shifting standards on evaluations of white subtype members. PMID- 14760968 TI - Gender role concerns in Estonian and Australian young adults. PMID- 14760969 TI - Zentralblatt fur Bakteriologie--100 years ago. The search for typhoid bacilli in drinking and river waters. PMID- 14760970 TI - Housekeeping enzymes as virulence factors for pathogens. AB - Housekeeping enzymes are ubiquitously present in almost all living beings to perform essential metabolic functions for the purpose of survival. These enzymes have been characterized in detail for many years. In recent years, there has been a number of reports indicating that some of these enzymes perform a variety of other functions. In case of many pathogens, certain enzymes play a role to enhance virulence. To perform such a function, enzymes must be located on the surface of pathogens. Although they do not have the typical signal sequence or membrane anchoring mechanisms, they do get secreted and are displayed on the surface, probably by their reassociation. Once on the surface, these enzymes interact with host components, such as fibronectin and plasminogen, or interact directly with the host cells, to trigger signal transduction and thereby enable the pathogens to colonize, persist and invade the host tissue. Therefore, certain housekeeping enzymes may act as putative virulence factors and targets for the development of new strategies to control the infection by using agents that can block their secretion and/or reassociation. PMID- 14760971 TI - Up-regulation of inducible nitric oxide synthase in Helicobacter pylori associated gastritis may represent an increased risk factor to develop gastric carcinoma of the intestinal type. AB - The enzyme inducible nitric oxide synthase (iNOS) is part of the host innate defense system against bacterial infection. During chronic inflammation, like that seen with a Helicobacter pylori infection, constant nitric oxide production may lead to tissue and DNA damage, thus increasing the patient's risk for developing cancer. Several investigations on iNOS expression in H. pylori associated gastritis have resulted in conflicting data. Therefore, we investigated the association between chronic H. pylori infection and iNOS expression in samples from stomach carcinoma patients as well as in antral biopsies from patients with H. pylori-associated gastritis. iNOS expression was analyzed by means of reverse transcriptase (RT)-PCR and quantified by competitive RT-PCR. To study in situ localization of iNOS in biopsy samples, immunohistochemistry was performed. iNOS enzyme activity was quantified using an arginine/citrulline assay. A significant increase in iNOS mRNA signal was only present in one-third of the analyzed patient biopsies with H. pylori-associated gastritis. These biopsies showed a 90% association with intestinal metaplasia and a 100% association with CagA-positive H. pylori. Intestinal metaplasia is discussed to be one step in the carcinogenesis of stomach cancer. Quantitation of iNOS transcripts and iNOS enzyme activity in non-cancerous mucosa of gastric cancer patients revealed a significant increase in iNOS transcripts and iNOS activity only in the mucosa of patients with stomach cancer of the intestinal type but not in the diffuse type. Our results support the hypothesis that CagA positive H. pylori strains are associated with the expression and activity of iNOS, and therefore might contribute to the development of intestinal metaplasia leading to gastric cancer of the intestinal type. PMID- 14760972 TI - Translocation of protein kinase-C with IP3-induced calcium mobilization by heat stable enterotoxin of Vibrio cholerae non-O1 in isolated rat enterocytes. AB - The activity of the calcium- and phospholipid-dependent enzyme protein kinase C (PKC) in response to heat-stable enterotoxin (NAG-ST) of Vibrio cholerae non-O1 was examined in isolated rat enterocytes. Optimal stimulation of the membrane bound PKC activity (about 4.3-fold) was observed after 1 min of incubation of cells with 10 ng/ml toxin; and the effects were dose dependent. Following NAG-ST treatment an increase in PKC activity in the membrane fraction was found with a concomitant decrease in the cytosolic fraction suggesting the redistribution of the enzyme. The pronounced enzyme activity in presence of a classical pseudosubstrate and its complete inhibition by Go 6976 suggested the involvement of a calcium-dependent isoform of PKC (PKC-alpha). A time course study employing an immunoblot assay provided evidence that NAG-ST led to almost complete translocation of PKC-alpha to the membrane. A 65% inhibition of enzyme activity in the membrane fraction and inhibition of its translocation to some extent by dantrolene treatment further suggested that the enzyme was translocated with the rise of intracellular calcium ([Ca2+]i). The phosphorylation of three membrane proteins by toxin-induced PKC in vitro and abolition of this phosphorylation by Go 6976 demonstrated that phosphorylation of these membrane proteins was PKC alpha mediated and might be involved in the alteration of membrane functions. PMID- 14760973 TI - Early detection of Borrelia burgdorferi sensu lato infection in Balb/c mice by co feeding Ixodes ricinus ticks. AB - In Europe, Borrelia burgdorferi is transmitted by Ixodes ricinus to animals and human. When infected and uninfected ticks co-feed on a host, spirochetes are transmitted from ticks to animal and also to uninfected ticks. Here, we used uninfected ticks to co-feed with infected ticks on mice to evaluate this method to detect early infection in mice. A total of 128 mice were challenged by infected nymphs placed in capsules glued on the back of the mice. Three days later uninfected larvae were added in the capsule to co-feed with infected nymphs and were examined for Borrelia infection after natural detachment. Infection in mice was also determined by xenodiagnosis and by spirochete isolation from ear skin biopsy and back skin biopsy taken at the tick attachment site one month after infection. A total of 111 mice were found to be infected by at least one of these four methods. Borrelia infection was observed in 95% of mice by the co feeding method, in 92% of mice by xenodiagnosis, in 69% and in 68% of mice by cultivation of ear and back skin biopsies, respectively. Our results demonstrate that the co-feeding method is a very sensitive method which can be used to detect very early infection in mice infected by tick bites. PMID- 14760974 TI - Mutations are involved in emergence of aminoglycoside-induced small colony variants of Staphylococcus aureus. AB - Staphylococcus aureus small colony variants (SCVs) occur frequently after local treatment with aminoglycosides and cause persistent as well as recurrent infections. So far, the molecular mechanism of the emergence of SCVs is not understood and regulatory as well as genetic mechanisms seem conceivable. To screen for possible mutations, the hemin biosynthetic gene cluster of a gentamicin-induced SCV was sequenced and was found to contain a deletion in the gene hemH. To further assess the influence of a high mutation rate on the development of SCVs, we tested the emergence of SCVs in a strain that had been inactivated in the DNA proofreading enzyme MutS. In the mutant, spontaneous SCVs emerged 556-fold more frequently than in the parent strain. By incubation in the presence of subinhibitory concentrations of gentamicin, the SCV frequency in the parent strain could be increased to 9.7 x 10(-6), whereas it remained rather stable in the mutant (1.8 x 10(-5)). Eighty percent of the gentamicin-induced SCVs were hemin auxotrophic in contrast to only 20% of the spontaneous SCVs which may explain the large proportion of hemin auxotrophs among clinical SCVs from patients previously treated with aminoglycosides. Additionally, a clinical S. aureus SCV isolate with a mutator phenotype, indicated by the generation of rifampicin-resistant mutants at a 16-fold higher frequency than in the reference strain S. aureus NCTC 8325, was characterized. The results demonstrate that a high mutation rate favours the emergence of SCVs, and suggest that mutations in general play an important role in the development of SCVs. PMID- 14760975 TI - Isolation of Helcococcus kunzii from a post-surgical foot abscess. AB - We report here a case in which a strain of Helcococcus kunzii was isolated from a foot abscess, in an immunocompetent patient, after surgery for metatarsal fracture. Correct species identification of the strain was readily obtained on the basis of morphological and biochemical features, together with 16S rRNA analysis. This case demonstrates that this species should be considered a potential pathogen in patients with no underlying disease. PMID- 14760976 TI - Chronic granulomatous lung infection caused by the dimorphic fungus Emmonsia sp. AB - A 64-year old farmer developed cough, dyspnoea on exertion, and recurrent febrile episodes. X-ray and CT scan revealed bilateral lower lobe opacities in his lungs. A transbronchial biopsy was performed and histopathological findings were interpreted as consistent with a pulmonary necrotizing clear-cell carcinoma and later as a Pneumocystis carinii pneumonia. Due to persistence of symptoms, six months later another lung biopsy was performed and a mould was cultured which was identified by 18S rDNA sequencing as Emmonsia sp. The patient showed some improvement under itraconazole treatment. This is the first description of a human infection with Emmonsia sp. in Germany. PMID- 14760977 TI - A review of conceptual models for assistive technology outcomes research and practice. AB - Conceptual models provide a theoretical basis for advancing scientific knowledge and improving professional practice. Although numerous assistive technology related models have appeared in the literature, there has been no systematic effort to assess them. Six conceptual models are reviewed here: Cook and Hussey's Human-Activity-Assistive Technology model; the World Health Organization's International Classification of Functioning, Disability, and Health; Scherer's Matching Person and Technology model; Gitlin's model of an AT user's "career"; social cognition decision-making theories; and Rogers' Perceived Attributes Theory. The models are reviewed in terms of six domains: background and goals; descriptive characteristics; indication of outcome measures; predictive characteristics; validation in the literature; and utility to assistive technology practitioners, developers, and consumers. The salient strengths and limitations are highlighted for each. Application of the models to advance theory, research, and practice is discussed. PMID- 14760978 TI - Cost analyses in assistive technology research. AB - Economic evaluation has become one of the principal methodologies in outcomes research within the health care field in general and specifically in assistive technology. Efforts to define and develop a consistent methodology for assistive technology economic evaluations have been hampered by lack of familiarity with the various terms and concepts associated with cost analysis, an essential aspect of economic evaluations. Adapting these concepts, which were constructed to suit a medical model, to the needs of the assistive technology field has been a challenge as well. This article outlines terms and concepts basic to cost analysis. The authors then consider five studies that relate costs to outcomes in order to illustrate the challenges, choices, and trade-offs researchers make when adapting this methodology to assistive technology. The article concludes by seeking to stimulate further discussion of the complexity inherent in assessing costs in assistive technology outcomes research and calling for the development of a standardized and consistent economic evaluation methodology. PMID- 14760979 TI - Sources of information about how to obtain assistive technology: findings from a national survey of persons with disabilities. AB - A modified discriminant function analysis was performed to determine the interaction between the source of information for assistive technology used by persons with disabilities. In the sample of 1,412 such persons, 901 were found to use some form of assistive technology in their daily lives. Ten distinct sources of information were specified. Respondents were able to mention up to three sources of information for each example of assistive technology used. A total of 930 sources were mentioned. Overall, the most mentioned information source was a physician or other health care professional, accounting for 53% of all sources mentioned. The only other frequently mentioned sources were family and friends (15%) and vocational rehabilitation counselors (13%). We found that physicians and health care professionals provide information to all groups without significant variance--this source of information is clearly nondiscriminant. Respondents who use family and friends as a source of information tend to be older, poorer, unemployed, more severely impaired, non-White, and, most significantly, render a more negative opinion of the amount and utility of information they have received about assistive technology, as well as the helpfulness of assistive technology in general. Persons obtaining information from vocational rehabilitation counselors tend to be better educated, non-White, unemployed, and have a more positive opinion about assistive technology. The limited size of the national population-based sample (n = 1,412) and very low number of responses indicating a state-based agency as the source of information about assistive technology did not allow state-by-state comparisons; alternative approaches, however, suggest a slight decline in the usage of the primary source (physician or other health care professional) with decreasing population, and a larger increase in the usage of family and friends as a source of information. PMID- 14760980 TI - Stroke rehabilitation: assistive technology devices and environmental modifications following primary rehabilitation in hospital--a therapeutic perspective. AB - The aim of this article is to describe the need for assistive devices and environmental modifications among long-living stroke survivors and to investigate if the need is continued and growing over time. The study sample of 155 consecutive stroke patients with stroke-related impairment, discharged home from three hospitals in Copenhagen from 1996 through 1998, constituted 20% of the total population of stroke survivors in this area. The results showed that 75% of these patients were provided with assistive devices and/or environmental modifications at discharge. Six months after discharge the proportion was 81%. The aids most frequently prescribed were bath seats, aids for mobility, grab bars, and removal of door thresholds. At follow-up 3-5 years later, 74% of the survivors were seen (76 patients). Almost all of the survivors were dependent on assistive devices and/or environmental modifications, most frequently wheelchairs and aids for walking and bathing. In addition there was a significant increase in aids for cooking/eating and reading/hearing/writing adaptations. Of those devices abandoned, most were aids for the household. These findings suggest that home visits by therapists should be required in order to target stroke survivors' changing needs for assistive devices and environmental modifications. PMID- 14760981 TI - Reliability of bench tests of interface pressure. AB - Determination of an appropriate wheelchair cushion to optimize loading on buttock tissue is crucial to pressure ulcer prevention. Standardized test methods aim to simplify selection by helping clinicians and users identify a class or category of cushions that will meet the important medical need of adequate pressure distribution. The objective of this project was to determine the test-retest reliability of interface pressure measurements taken using bench tests as opposed to human subject tests. Ten wheelchair cushions were tested following the methods for interface pressure measurement as defined in a draft International Organization for Standardization document. Dispersion index, contact area, percent force in the ischial regions, peak pressure index, and seating pressure index-standard deviation are reliable measures. Average pressure is reliable but not very volatile between cushions. The data also indicate that peak pressure, seating pressure index-skew (SPI-sk), and the other five percent force regions are not reliable. Certain bench interface pressure variables were found to have adequate intralaboratory repeatability. Interlaboratory reliability must also be tested. If a bench interface pressure test is used to indicate cushion performance, its validity should also be studied. Research is underway to relate interface pressure variables to clinical measurements of wheelchair users. Once validity is shown, standardized test results can then be used by clinicians to simplify and improve the wheelchair cushion selection process. PMID- 14760982 TI - Powered tilt/recline systems: why and how are they used? AB - Prolonged static sitting can lead to discomfort, pain, pressure sores, spinal curvatures, and loss of functional independence. In order to counteract these harmful effects, adjustable tilt and/or recline systems are often prescribed. Considering the current context of assistive technology service delivery and budget cuts, it is essential to have a better knowledge of the use of these technical aids and user's satisfaction with them. The purpose of this study was to characterize the use of powered tilt and recline systems. A questionnaire was developed for this purpose, and 40 subjects were interviewed at home. They were asked to identify, from a list of 25 objectives, the reasons for which they used their repositioning system and to rank these reasons in order of importance. For each objective, they were also asked to identify the frequency and range of use as well as their satisfaction level with their system. Results revealed that 97.5% of the subjects were using their powered tilt and recline system everyday, and their satisfaction was high. The main objectives for using this type of assistive technology were to increase comfort and to promote rest. Although mainly descriptive, results are of clinical relevance and can be helpful when selecting wheelchairs. PMID- 14760983 TI - Accomplishing technology transfer: case-based lessons of what works and what does not. AB - This paper presents lessons drawn from technology transfer case studies that address the persistent question: "What works, what does not, and why?" Each lesson highlights critical factors determining success or failure and is substantiated by case studies that exemplify the lesson. The case examples involve either the commercialization of prototype inventions (supply-push technology transfer) or the acquisition of desired technologies from other fields of application (demand-pull technology transfer). The cases present the chronology of events as they actually occurred, including supporting information from the other participants. Applying the lessons should help avoid common mistakes while increasing the likelihood of accomplishing the desired outcomes. PMID- 14760984 TI - Efficacy of endoventricular circular patch plasty: will this procedure improve the prognosis of patients with ischemic cardiomyopathy? AB - OBJECTIVES: We report hemodynamic and clinical results of our series of endoventricular circular patch plasty (Dor operation) and consider some advantages of this procedure for patients with ischemic cardiomyopathy. METHODS: Between 1996 and 2001, 23 consecutive patients with left ventricular aneurysm and/ or ischemic cardiomyopathy after myocardial infarction who underwent Dor operation were included in this study. Hemodynamic and clinical results of Dor operation were analyzed periodically. Patients were divided into two groups according to the extent of asynergy, more than 60% or not, i.e., an ischemic cardiomyopathy group (ICM group) or a simple left ventricular aneurysm group (sLVA group). RESULTS: Hospital mortality was 4.4%. Postoperative New York Heart Association functional class was improved in all survivors to class I or II. Postoperative ejection fraction (EF) increased and postoperative left ventricular (LV) volume decreased in all survivors. In both groups, early and 1 year postoperative EF increased significantly. Additionally, end-diastolic and end systolic volumes decreased significantly in the early postoperative period. Postoperative LV volume had re-enlarged in the cases in which preoperative left ventricular end-systolic volume index was more than 90 mL/m2, though left ventricular ejection fraction was maintained or rather improved at 1 year postoperatively. The survival rates after 3 years of the operation in the sLVA and ICM groups were 85.7% and 81.3%. CONCLUSION: Though patients with ischemic cardiomyopathy with severe LV dysfunction may benefit the most from Dor operation, postoperative LV re-dilatation may deteriorate late mortality. At operation, whether optimal LV size and shape can be reconstructed and the timing of operation are the important issues because they affect prognoses. PMID- 14760985 TI - Redo coronary artery bypass grafting: early and mid-term results. AB - OBJECTIVES: Redo coronary artery bypass grafting (CABG) has been gradually increasing in Japan. We prospectively collected redo-CABG data and evaluated these the early and remote results. METHODS: Between 01/01/1994 and 06/30/2002, a total of 71 patients underwent isolated redo-CABG in our hospital group. The interval between operations was 7.8 +/- 6.1 years. Previous surgery was CABG in all patients. Perioperative, early angiographic, and follow-up results were analyzed. RESULTS: The mean number of grafts was 2.9 +/- 1.2. There were 4 incidences of injury to the heart or graft during sternal re-entry or during dissection of the heart. There was 1 hospital death (2.8%) and 19 major complications (26.8%), including 7 patients (9.9%) with postoperative congestive heart failure and 2 (2.8%) with postoperative myocardial infarction. Postoperative angiography was obtained in 47 patients and their overall stenosis free patency rate was 93.9%. Follow-up was completed for all hospital survivors with a mean follow-up of 3.9 +/- 2.2 years. The event-free and survival rates at 5 years were 76.4% and 83.9%, respectively. CONCLUSION: In our limited experience, redo-CABG was performed with acceptable risks and its long-term results were satisfactory. PMID- 14760986 TI - Noonan syndrome associated with anomalous coronary artery and other cardiac defects. AB - This case-report describes an 11-month-old infant, who had been diagnosed with Noonan syndrome with a variety of associated anomalies. Multiple cardiac anomalies were present, consisting of dysplastic pulmonary valve, symmetric biventricular hypertrophy, atrial septal defect and right ventricular outflow tract (RVOT) obstruction, in which the pressure gradient measured 73 mmHg, and anomalous coronary artery. Systolic anterior motion of the mitral valve (SAM) was present, without remarkable clinical significance. RVOT transannular repair with non-cusped xenograft along with resection of hypertrophied right ventricular outflow myocardium. Left ventricular outflow tract (LVOT), which had no clinical sign of obstruction, was left untouched, expecting the RVOT repair also effectively release LVOT dynamic obstruction. The postoperative echocardiography revealed residual SAM without significant pressure gradient through LVOT. PMID- 14760988 TI - Mitral valve replacement through right thoracotomy after previous coronary artery bypass grafting: the usefulness of brachial artery cannulation, perfused ventricular fibrillation with moderate hypothermia, and minimal dissection techniques. AB - It has been reported by several authors that a right thoracotomy for mitral valve surgery can be useful after previous coronary aortery bypass grafting (CABG). A 76-year-old man with mitral valve regurgitation after previous CABG underwent mitral valve replacement with some modified techniques. Cardiopulmonary bypass was established with right brachial artery cannulation and right femoral venous cannulation with the aid of vacuum-assisted venous drainage. Ventricular fibrillation (VF) was induced by rapid pacing of the ventricle, and mitral valve replacement was performed under perfused VF with moderate hypothermia. The patient's postoperative course was uneventful. This method appears to be a safe and easy alternative mitral valve surgery for complicated cases of this type. PMID- 14760987 TI - Lung adenocarcinoma with coexisting pulmonary cryptococcoma. AB - A 73-year-old female was referred to our hospital in June 2000 for the evaluation of an abnormal shadow in the left upper lobe of her lung and two other shadows in the left lower lobe. A computed tomography examination revealed an ill-defined tumor with ground glass opacity in the left upper division segment, suggesting a primary lung adenocarcinoma, and two well-defined nodules in the left lower lobe, suggesting metastatic carcinomas. A transbronchial lung biopsy of the tumor in the upper division segment confirmed the presence of an adenocarcinoma but failed to provide a histological diagnosis for the nodules in the lower lobe. A partial resection of the left lower lobe using video-assisted thoracoscopy revealed granulomatous tissue with no signs of malignancy. A curative resection of the adenocarcinoma in the left upper division segment was therefore performed. Histologically, the resected specimens obtained from the left lower lobe were diagnosed as cryptococcomas. The patient was diagnosed as having a stage IA lung adenocarcinoma and two cryptococcomas. The patient is presently well and has not experienced any recurrences or relapse for 3 years since the resection. PMID- 14760989 TI - Primary thymic adenocarcinoma with production of carbohydrate antigen 19-9 and carcinoembryonic antigen. AB - A 59-year-old male, whose chest X-ray showed an abnormal shadow, visited us for further study. Laboratory examination showed the elevated level for both carbohydrate antigen 19-9 (CA19-9) and carcinoembryonic antigen (CEA) in serum, and the chest X-ray and computed tomography showed an anterior mediastinal mass. Under a diagnosis of thymic malignancy, extended thymectomy with combined resection of the pleura and pericardium was performed. Histopathological findings showed a well-differentiated adenocarcinoma of the thymus, in which CA19-9 and CEA were positive immuno-histochemically. The level of serum CA19-9 and CEA returned to normal ranges postoperatively, however, the tumor recurred in local site with re-elevation of these tumor markers at the 20th month after surgery. He died at 4th month after the first recurrence despite the intensive chemotherapy. We report an extremely rare case of primary thymic adenocarcinoma with the production of CA19-9 and CEA. PMID- 14760990 TI - Pneumothorax as a presenting manifestation of early sarcoidosis. AB - Pneumothorax rarely develops sarcoidosis. A 21-year-old man with early sarcoidosis presenting as pneumothorax is reported. The patient came to our institute with severe chest pain and dyspnea. Plain chest roentgenograms revealed pneumothorax in the left lung. A chest tube was inserted to inflate the lung. Subsequent computed tomography demonstrated subpleural blebs in the upper lobe of the left lung. Continuous treatment with tube drainage was performed. However, surgical intervention was needed since long-term tube drainage turned out to be unsuccessful. Thoracoscopic partial extirpation on the left upper lobe was performed. The histology of the obtained lung tissue showed non-caseating granulomas composed of epithelioid cells and occasional giant cells. A diagnosis of sarcoidosis was made on the basis of the histological report. Although early sarcoidosis presenting as pneumothorax is rare in young patients, the possibility of a sarcoidosis should be considered. PMID- 14760991 TI - Synchronous double cancers developing from the wall of bullae in the bilateral lungs. AB - A rare case of synchronous double lung cancers having developed from bilateral bullous disease is reported. A 51-year-old man was admitted because of severe cough. Imaging studies revealed a left apical bulla measuring 10 cm, and another bulla measuring 8 cm containing viscous fluid on the right apex. In the next year, chest computed tomography showed increased size of the mass in the right apical bulla. Upper right lobectomy and left bullectomy were performed. Histological examination of the resected specimens revealed a large cell carcinoma having developed the wall of the bulla in the right apex, and a moderately differentiated papillary adenocarcinoma having developed in the wall of the left bulla. The patient had an uneventful recovery and has been in good health without recurrence for 3 years since surgery. We emphasize the need to be aware of the potential development of lung cancer in patients with bullous disease. PMID- 14760992 TI - Metastasis of thyroid cancer to primary lung cancer. AB - We present a case of a 65-year-old woman whose thyroid cancer metastasized to the lesion of primary lung cancer. Ten years after total thyroidectomy for thyroid cancer, chest radiograph by medical check-up demonstrated three nodular lesions in the bilateral lung fields. Segmental resection of the left S6, partial resection of right S4 and left S10 were performed to remove those lesions. Histologically, small nodules in the right S4 and S10 were diagnosed as a metastatic tumor of thyroid and well differentiated adenocarcinoma, respectively. Left S6 lesion 1.5 cm in diameter was also diagnosed as well-differentiated adenocarcinoma (Noguchi type C), however, small metastatic foci of papillary adenocarcinoma was identified within the lesion which revealed to be "cancer in cancer metastasis". Metastasis of cancer to another primary cancer is a rare event. We discuss interesting phenomenon of cancer in cancer metastasis with a review of the literature. PMID- 14760993 TI - Subepicardial aneurysm following ventriculotomy closure of ventricular septal rupture due to acute myocardial infarction. AB - We report an unusual case of subepicardial aneurysm (SEA) of the left ventricle. An 82-year-old man had undergone patch closure of a ventricular septal rupture due to anterior acute myocardial infarction. A postoperative left ventriculogram showed the presence of contrast medium outside the left ventricle, and urgent surgery was performed. The lesion was diagnosed as SEA, and was repaired before rupture. PMID- 14760994 TI - Positive end-expiratory pressure depressed cardiovascular autonomic nervous system activity in acute brain damaged rabbits under general anesthesia. AB - Artificial ventilation with positive end-expiratory pressure (PEEP) is commonly applied for brain damaged patients. However, the effect of the ventilation on brain function, including cardiovascular autonomic nervous system (CANS) activity, is not well elucidated. In order to investigate the effect of 5 cmH2O PEEP on CANS activity in brain damaged rabbits under general anesthesia, we produced acute brain damage by intracranial balloon inflation. Measurements were made before (control) and after application of PEEP, and after inflation with incremental volume of the balloon. Power spectral analyses of heart rate variability (HRV) and systolic arterial pressure variability (SAPV) were used for the assessment of CANS activity. Spectral powers in the low-frequency range of 0.04 to 0.40 Hz (LF) and high-frequency range of 0.75 to 1.40 Hz (HF) were computed, and their ratio LF/HF was assessed as the neural balance of CANS. The animals in group P were ventilated with 5 cmH2O PEEP, while those in group Z were ventilated with zero end-expiratory pressure. Colored microsphere counting was used for the assessment of brain circulation. In the results, PEEP had no effect on HRV and SAPV parameters before induction of brain damage. After inflation with incremental volume of the balloon, log (HF) and log (LF) in group P were lower than in group Z in HRV analysis, and log (LF) in group P was lower than in group Z in SAPV analysis. Microsphere counting revealed that brain blood flow was reduced during the progression of brain damage and showed a significant difference after application of PEEP between the groups. We concluded that 5 cmH2O PEEP depressed CANS activity during the progression of brain damage in rabbits and that this was partly due to aggravated brain function induced by PEEP. PMID- 14760995 TI - Need for health promotion based on evidence in cross sectional school health data. AB - With the purpose of evaluating the health condition of children in public primary schools in the province of Cordoba in Argentina, cross sectional data were collected from the School Health Program of the Ministry of Health of Cordoba Province in 2001. To conduct an analysis, a total of 25,701 pupils of the first and the seventh grades from one hundred and fifty primary schools were studied. The data consisted of variables in three groups: i.e., nutrition, vaccination and disease groups. Descriptive statistics revealed a higher prevalence in scoliosis, lower weight, no scar from BCG vaccination and skin transmission diseases in urban and rural areas than in interurban areas, while male genital diseases showed a high prevalence in the three areas. Dental caries had a high prevalence in rural areas. Factor analysis was carried out and six factors were extracted out of 27 variables to cover 64 % of total variance. Regression analyses revealed that nutritional variables such as lower weight and the absence of BCG vaccination were significantly related to scoliosis, and that lower weight significantly affected skin transmission diseases. We considered the necessity of modifying conduct in the school health program from the viewpoints of health promotion and community participation. PMID- 14760996 TI - Some approaches to treatment of patients with thyroid nodular diseases in the Semipalatinsk region of Kazakhstan. AB - It was determined by experts that in the region adjacent to the Semipalatinsk Nuclear Test Site thyroid nodular prevalence was significantly associated with radiation dose from nuclear weapons testing. The medical rehabilitation of patients is of special practical and scientific importance in this region. Some patients have contraindications for surgery and radioiodine or refuse them for various reasons. Percutaneous intranodular injection therapy by "Paoscle" (PIITP) was used for treatment of patients with benign thyroid nodular diseases. The study group included a total of 107 patients (mainly "pretoxic" and "compensated" nodules). Seventy-four patients received PIITP on an out-patient basis. Seventy two of them were women and 2 men, mean age, 52.9 +/- 1.3 years; range 26 to 77 years. Thyroid ultrasound examination, fine-needle aspiration biopsy, thyroid function tests and cytopathology were performed in all patients to evaluate the effectiveness of treatment. The nodule volume reduction rate for patients with thyroid adenomas was 56 % on average and 60.2 % in the case of colloid nodules. The reduction rate of thyroid adenomas with necrotic and cystic degeneration was higher and reached 72 %. The suggested method is indicated for cases of benign nodular thyroid diseases (cysts, adenomas, adenomas with necrotic and cystic degeneration, colloid nodules, polynodular goiter). We did not reveal any complications or thyroid test abnormalities after the treatment. A tendency to normalization of the blood serum thyroglobulin level and antibodies to thyroglobulin was documented. Analysis of the treatment results revealed that this approach is effective, inexpensive, safe, well tolerated and can be used on an outpatient basis. PMID- 14760997 TI - A case of venous angioma with arteriovenous shunts--case report. AB - A 35-year-old man presented with a sudden headache and disturbance of consciousness. On admission, his consciousness level was Japan Coma Scale 100. Computed tomography disclosed a subarachnoid hemorrhage (SAH) and right cerebellar hematoma. Angiography was performed and, at first, arteriovenous malformation of the posterior fossa was diagnosed. Then external decompression of the posterior fossa and ventricular drainage were performed, followed by barbiturate therapy. Repeat angiography revealed that the lesion was a venous angioma with arteriovenous shunts. On day 37, subtotal removal of the lesion was performed. Intraoperatively, acute brain swelling emerged and partial internal decompression of the right cerebellar hemisphere was performed. The postoperative course was comparatively good and the patient was discharged with very mild ataxia. The patient is now being followed up in our outpatient clinic. PMID- 14760998 TI - The intertwining of politics and science in psychoanalytic history. PMID- 14760999 TI - The good fight: psychoanalysis in the age of managed care. PMID- 14761000 TI - Standards and standardization. PMID- 14761001 TI - Thoughts on authority and leadership. PMID- 14761002 TI - Psychoanalytic discourse at the turn of our century: a plea for a measure of humility. PMID- 14761003 TI - Can we integrate the diverse theories and practices of psychoanalysis? PMID- 14761004 TI - A plea for constructive dialogue. PMID- 14761006 TI - Psychoanalytic theory as a form of countertransference. PMID- 14761007 TI - The forbidden quest and the slippery slope: roots of authoritarianism in psychoanalysis. PMID- 14761008 TI - The lawsuit revisited. PMID- 14761009 TI - The history of lay analysis: emendations. PMID- 14761010 TI - The lawsuit from the plaintiffs' perspective. PMID- 14761011 TI - Toward a psychoanalytic politics. PMID- 14761012 TI - A brief history of psychoanalysis in Argentina. PMID- 14761013 TI - IMRT may be used to excess because of its higher reimbursement from Medicare. For the proposition. PMID- 14761014 TI - IMRT may be used to excess because of its higher reimbursement from Medicare. Against the proposition. PMID- 14761015 TI - Grangeat-type helical half-scan computerized tomography algorithm for reconstruction of a short object. AB - Currently, cone-beam computerized tomography (CT) and micro-CT scanners are under rapid development for major biomedical applications. Half-scan cone-beam image reconstruction algorithms assume only part of a scanning turn, and are advantageous in terms of temporal resolution and image artifacts. While the existing half-scan cone-beam algorithms are in the Feldkamp framework, we have published a half-scan algorithm in the Grangeat framework for a circular trajectory [Med. Phys. 30, 689-700 (2003)]. In this paper, we extend our previous work to a helical case without data truncation. We modify the Grangeat's formula for utilization and estimation of Radon data. Specifically, we categorize each characteristic point in the Radon space into singly, doubly, triply sampled, and shadow regions, respectively. A smooth weighting strategy is designed to compensate for data redundancy and inconsistency. In the helical half-scan case, the concepts of projected trajectories and transition points on meridian planes are introduced to guide the design of weighting functions. Then, the shadow region is recovered via linear interpolation after smooth weighting. The Shepp Logan phantom is used to verify the correctness of the formulation, and demonstrate the merits of the Grangeat-type half-scan algorithm. Our Grangeat type helical half-scan algorithm is not only valuable for quantitative and/or dynamic biomedical applications of CT and micro-CT, but also serves as an intermediate step towards solving the long object problem. PMID- 14761016 TI - Magnetic collimation and metal foil filtering for electron range and fluence modulation. AB - We investigated the use of magnetically collimated electron beams together with metal filters for electron fluence and range modulation. A longitudinal magnetic field collimation method was developed to reduce skin dose and to improve the electron beam penumbra. Thin metal foils were used to adjust the energies of magnetically collimated electrons. The effects for different types of foils such as Al, Be, Cu, Pb, and Ti were studied using Monte Carlo calculations. An empirical pencil beam dose calculation model was developed to calculate electron dose distributions under magnetic collimation and foil modulation. An optimization method was developed to produce conformal dose distributions for simulated targets such as a horseshoe-shaped target. Our results show that it is possible to produce an electron depth dose enhancement peak using similar techniques of producing a spread-out Bragg peak. In conclusion, our study demonstrates new aspects of using magnetic collimation and foil filtration for producing fluence and range modulated electron dose distributions. PMID- 14761017 TI - Search for lesions in mammograms: statistical characterization of observer responses. AB - We investigate human performance for visually detecting simulated microcalcifications and tumors embedded in x-ray mammograms as a function of signal contrast and the number of possible signal locations. Our results show that performance degradation with an increasing number of locations is well approximated by signal detection theory (SDT) with the usual Gaussian assumption. However, more stringent statistical analysis finds a departure from Gaussian assumptions for the detection of microcalcifications. We investigated whether these departures from the SDT Gaussian model could be accounted for by an increase in human internal response correlations arising from the image-pixel correlations present in 1/f spectrum backgrounds and/or observer internal response distributions that departed from the Gaussian assumption. Results were consistent with a departure from the Gaussian response distributions and suggested that the human observer internal responses were more compact than the Gaussian distribution. Finally, we conducted a free search experiment where the signal could appear anywhere within the image. Results show that human performance in a multiple-alternative forced-choice experiment can be used to predict performance in the clinically realistic free search experiment when the investigator takes into account the search area and the observers' inherent spatial imprecision to localize the targets. PMID- 14761018 TI - Measurement of renal extraction fraction using contrast-enhanced computed tomography. AB - Renal extraction fraction (EF) is the percentage of plasma entering the glomerulus which is filtered. Contrast agents which are freely filtered and neither secreted nor reabsorbed, may be used as markers for renal filtration, allowing EF to be calculated from computed tomography (CT) measurements of systemic vessels and renal veins. CT scans of 10 adult patients having no known renal disease were studied in this manner, giving EF values averaging 12.6% and 12.3% for the right and left kidneys, respectively, compared to the accepted value of 15%-20%. EF measurement using CT may provide noninvasive evaluation of renal function, complementing CT-derived morphologic information. PMID- 14761019 TI - Image filtering for improved dose resolution in CT polymer gel dosimetry. AB - X-ray computed tomography (CT) has been established as a feasible method of performing dosimetry using polyacrylamide gels (PAGs). A small density change occurs in PAG upon irradiation that provides contrast in PAG CT images. However, low dose resolution limits the clinical usefulness of the technique. This work investigates the potential of using image filtering techniques on PAG CT images in order to reduce image noise and improve dose resolution. CT image noise for the scanner and protocol used for the gel images is analyzed and found to be Gaussian distributed and independent of the contrast level in the images. As a result, several filters for reducing spatially invariant noise are investigated: mean, median, midpoint, adaptive mean, alpha-trimmed mean, sigma mean, and a relatively new filter called SUSAN (smallest univalue segment assimilating nucleus). All filters are applied, using 3x3, 5x5, and 7x7 pixel masks, to a CT image of a PAG irradiated with a stereotactic radiosurgery dose distribution. The dose resolution within 95% confidence (D(delta)95%) is calculated and compared for each filtered image, as well the unfiltered image. In addition, the ability of the filters to maintain the spatial integrity of the dose distribution is evaluated and compared. Results clearly indicate that the filters are not equal in their ability to improve D(delta)95% or in their effect on the spatial integrity of the dose distribution. In general, increasing mask size improves D(delta)95% but simultaneously degrades spatial dose information. The mean filter provides the greatest improvement in D(delta)95%, but also the greatest loss of spatial dose information. The SUSAN, mean adaptive, and alpha-trimmed mean filters all provide comparable, but slightly poorer dose resolution. In addition, the SUSAN and adaptive filters both excel at maintaining the spatial distribution of dose and overall are the best performing filters for this application. The midpoint filter, normally useful for Gaussian noise, is poor all-round, dramatically distorting the dose distribution for masks greater than 3x3. The median filter, a common edge preserving noise reduction filter, performs moderately well, but artificially increases high dose gradients. The sigma filter preserves the spatial distribution of dose very well but is least effective at improving dose resolution. In summary, dose resolution can be significantly improved in CT PAG dosimetry through postprocessing of CT images using spatial noise reduction filters. However, such filters are not equal in their ability to improve dose resolution or to maintain the spatial integrity of the dose distribution and an appropriate filter must be chosen depending on clinical demands of the application. PMID- 14761020 TI - Comparison of p-type commercial electron diodes for in vivo dosimetry. AB - This paper compares the characteristics of three types of commercial p-type electron diodes specially designed for in vivo dosimetry (Scanditronix EDD2, Sun Nuclear QED 111200-0 and PTW T60010E diodes coupled with a Therados DPD510 dosimeter) in electron fields with energies from 4.5 to 21 MeV, and in conditions similar to those encountered in radiotherapy. In addition to the diodes, a NACP plane parallel ionization chamber and film dosimeters have been used in the experiments. The influence of beam direction on the diode responses (directional effect) was investigated. It was found to be the greatest for the lowest electron beam energy. At 12 MeV and an incidence of +/- 30 degrees, the variation was found to be less than 1% for the Scanditronix and Sun Nuclear diodes and less than 3% for the PTW one. The three diodes exhibited a variation in sensitivity with dose-per-pulse of less than 1% over the range 0.18-0.43 mGy/pulse. The temperature dependence was also studied. The response was linear for the three diodes between 22.2 and 40 degrees C and the sensitivity variations with temperature were (0.25+/-0.01)%/degree C, (0.28+/-0.01)%/degree C, and (0.02 +/ 0.01)%/degree C for Scanditronix, Sun Nuclear, and PTW diodes, respectively. Finally the perturbation to the irradiation field induced by the presence of diodes placed at the surface of a homogeneous phantom was investigated and found to be significant, both at the surface and at the depth of maximum dose (several tens of percent) for all three diode types. There is an increase of dose right underneath the diode (close to the surface) and a dose shadow at the depth of maximum. The study shows that electron diodes can be used for in vivo dosimetry provided their characteristics are carefully established before use and taken into consideration at the time of interpretation of the results. PMID- 14761021 TI - Positronium: review of symmetry, conserved quantities and decay for the radiological physicist. AB - The PET-CT scanner, which overlays a high spatial resolution CT image on a relatively low resolution physiologic PET image, has greatly increased the efficacy of the PET imaging modality. With the resulting increase in the presence of PET imaging in the radiologic and medical physics communities, medical physicists can expect professional responsibilities in acceptance testing, quality assurance, radiation safety and resident teaching of this new modality. With this in mind we present a review of a subtle aspect of PET: the formation, symmetry properties and decay of positronium; the hydrogen like positron-electron complex which forms prior to annihilation. PMID- 14761022 TI - Suppression of intensity transition artifacts in statistical x-ray computer tomography reconstruction through radon inversion initialization. AB - Statistical reconstruction (SR) methods provide a general and flexible framework for obtaining tomographic images from projections. For several applications SR has been shown to outperform analytical algorithms in terms of resolution-noise trade-off achieved in the reconstructions. A disadvantage of SR is the long computational time required to obtain the reconstructions, in particular when large data sets characteristic for x-ray computer tomography (CT) are involved. As was shown recently, by combining statistical methods with block iterative acceleration schemes [e.g., like in the ordered subsets convex (OSC) algorithm], the reconstruction time for x-ray CT applications can be reduced by about two orders of magnitude. There are, however, some factors lengthening the reconstruction process that hamper both accelerated and standard statistical algorithms to similar degree. In this simulation study based on monoenergetic and scatter-free projection data, we demonstrate that one of these factors is the extremely high number of iterations needed to remove artifacts that can appear around high-contrast structures. We also show (using the OSC method) that these artifacts can be adequately suppressed if statistical reconstruction is initialized with images generated by means of Radon inversion algorithms like filtered back projection (FBP). This allows the reconstruction time to be shortened by even as much as one order of magnitude. Although the initialization of the statistical algorithm with FBP image introduces some additional noise into the first iteration of OSC reconstruction, the resolution-noise trade-off and the contrast-to-noise ratio of final images are not markedly compromised. PMID- 14761023 TI - Boron neutron capture therapy of skin melanomas at the RA-6 reactor: a procedural approach to beam set up and performance evaluation for upcoming clinical trials. AB - This article reports on the progress of the modeling and experimental characterization of the RA-6 reactor neutron beam, designed for the upcoming BNCT clinical trials of skin melanoma, and presents the first theoretical analysis of such beam performance. The aspects relating to surface source modeling and assessment, beam dosimetry, treatment planning system calibration, and treatment planning optimization are presented herein. Several methods and criteria were established in order to provide guidance for future clinical studies conducted in this facility. Following a realistic model, the theoretical analysis was based on a clinical case of malignant melanoma in extremities. Owing to the complex geometry of the tumor, this particular clinical case represents one of the most difficult lesions to be treated. This article discusses the thorough evaluation stage that has led to the optimization of the treatment planning procedure. Two candidate plans were proposed, and dose-volume distributions in the target volume were evaluated on the basis of the application of a series of criteria that define the critical normal structures which limit the dose delivered. In spite of the complexity of the clinical case under review, results showed that only 4% of the tumor volume is underdosed in cases of mean blood 10B concentration values, even in the most unfavorable analysis. The overall results suggest that this BNCT facility is prepared to rigorously explore the clinical efficacy of the RA-6 beam and the BNCT treatment modality for peripheral melanomas. PMID- 14761024 TI - Estimating three-class ideal observer decision variables for computerized detection and classification of mammographic mass lesions. AB - We are using Bayesian artificial neural networks (BANNs) to classify mammographic masses in schemes for computer-aided diagnosis, and we are extending this methodology to a three-class classification task. We investigated whether a BANN can estimate ideal observer decision variables to distinguish malignant, benign, and false-positive computer detections. Five features were calculated for 63 malignant and 29 benign computer-detected mass lesions, and for 1049 false positive computer detections, in 440 mammograms randomly divided into a training and testing set. A BANN was trained on the training set features and applied to the testing set features. We then used a known relation between three-class ideal observer decision variables and that used by a two-class ideal observer when two of three classes are grouped into one class, giving one decision variable for distinguishing malignant from nonmalignant detections, and a second for distinguishing true-positive from false-positive computer detections. For comparison, we grouped the training data into two classes in the same two ways and trained two-class BANNs for these two tasks. The three-class BANN decision variables were essentially identical in performance to the specifically trained two-class BANNs, with the average difference in area under the ROC curves being less than 0.0035 and no differences in area being statistically significant. Thus, the BANN outputs obey the same theoretical relationship as do the three class and two-class ideal observer decision variables, which is consistent with the claim that the three-class BANN output can provide good estimates of the decision variables used by a three-class ideal observer. PMID- 14761025 TI - Comparison of beam characteristics of a gold x-ray target and a tungsten replacement target. AB - A new W-Cu target was designed to replace the existing Au target on a linear accelerator model in common use in radiotherapy. This work shows that targets of different material composition can be designed to produce beams with the same dosimetric character over a wide range of beam energies without adjusting the beam energy. The target design objective was to improve mechanical robustness, replacing water in the beam path with a Cu heat sink, without altering the beam properties for the nominal clinical energy range of 4-25 MV. The W-Cu could then be installed in place of the Au target without recommissioning. The effect of the target swap was measured in the test cells for 11 different beams ranging in nominal energy from 4 to 25 MV, with focus on open field dose distributions, including diagonal profiles taken for the largest (40x40 cm) field, measured at 4 different gantry angles. Depth dose curves agreed to 0.4% or better, profiles to 1.2% or better. Monte Carlo simulations of the treatment head were done for representative energies of 6 and 18 MV. Calculated and measured dose distributions generally matched within 1%, although dose measured in the build-up region of large fields was significantly more than in the simulations. Calculated spectral distributions on the central axis and angular distributions of energy fluence matched for the two targets, whereas angular distributions of fluence were significantly different. Matching energy fluence gave a more favorable match of dose profiles than matching fluence. The target was further tested on several machines operating in a radiotherapy clinic. Measurements were made for a wide range of open field sizes and with selected wedges and blocks. Dose distributions for the two targets agreed to 1.4% or better, including the dose in wedged fields. Wedge factors changed by no more than 0.5%, transmission through a 4.4 HVL block no more than 1.5%. The response of the monitor chamber was found to change, generally by 1%-2%. Therefore, when the W-Cu target was used to replace the Au target, the output of the machine was measured and adjusted appropriately, but there was no requirement for recommissioning. PMID- 14761026 TI - A-Si:H/CsI(Tl) flat-panel versus computed radiography for chest imaging applications: image quality metrics measurement. AB - Amorphous silicon (a-Si:H) flat-panel (FP) imaging systems have recently become commercially available for both chest and mammographic imaging applications. It has been shown that this new detector technology offers better image quality and various operational advantages over the computed radiography (CR) which to date has been the most widely implemented and used digital radiography technique. However, most image quality measurements reported on flat-panel systems have been performed on prototype systems in laboratories while those for CR systems were typically independently performed and reported on in separate studies. To directly compare the two technologies, we have measured the image properties for a commercial amorphous silicon/cesium iodide [a-Si:H/CsI(Tl)] flat-panel based digital chest system and a commercial CR system under clinical imaging conditions. In this paper, measurements of image quality metrics, including the modulation transfer functions (MTFs), noise power spectra (NPSs), and detective quantum efficiencies (DQEs), for the FP and CR systems are presented and compared. Methods and issues related to these measurements are discussed. The results show that the flat-panel system has slightly lower MTF but significantly higher DQEs than the CR system. The DQEs of the flat-panel system were found to increase with the exposure while those of the CR system decrease slightly with the exposure. PMID- 14761027 TI - Determining optimal two-beam axial orientations for heart sparing in left-sided breast cancer patients. AB - BACKGROUND AND PURPOSE: The optimal intensity fluence profile of a beam depends on the profiles of other beams but most optimizations assume fixed beam orientations, a priori. Breast cancer radiotherapy attempts to cover the target and to spare critical structures such as the heart and lungs. The study aims are (1) to determine and document the optimal two-beam orientation that best spares the heart for left-sided breast cancer patients and (2) to investigate the influence of the treatment technique (i.e., conformal versus intensity modulation) on the optimal objective cost function. MATERIAL AND METHODS: Ten left-sided breast cancer patients were planned using a conformal (3DCRT) and a simplified intensity modulated (sIMRT) technique using predefined segments and different two-beam orientations. Optimal segment weights were determined exhaustively for all axial two-beam combinations, in 5 degree increments, by minimizing a quadratic objective cost function. The resulting objective cost function was analyzed with respect to target geometry and treatment technique. RESULTS: The sIMRT plans are generally less sensitive to beam orientation compared to 3DCRT plans. Optimal two-beam orientations for 3DCRT and sIMRT plans exist and they correspond to a hinge angle of approximately 188 degrees and 160 degrees or 210 degrees (the latter is bimodal), respectively. CONCLUSIONS: The optimization software is a useful tool that can test many different beam combinations and estimate their associated objective cost values. Afterwards, the most promising beam orientations could be re-optimized under the TPS to fine-tune and verify the dose distributions. Optimal uniform two-beam orientations for the breast consist of opposing tangential medial and lateral beams. Optimal nonuniform two-beam orientations for left-sided breast cancers are bimodal, containing hinge angles around 160 degrees and 210 degrees. Nonuniform beam techniques are less sensitive to beam orientation compared to uniform beam techniques and result in significantly improved heart sparing but at a cost of slightly compromised planning target volume coverage. PMID- 14761028 TI - Comparison between manual and automatic segment generation in step-and-shoot IMRT of prostate cancer. AB - PURPOSE: To compare two methods to generate treatment plans for intensity modulated radiotherapy (IMRT) of prostate cancer, delivered in a step-and-shoot mode. The first method uses fluence optimization (inverse planning) followed by conversion of the fluence weight map into a limited number of segments. In the second method, segments are manually assigned using a class solution (forward planning), followed by computer optimization of the segment weights. METHODS: Treatment plans for IMRT, utilizing a simultaneous integrated boost, were created. Plans comprise a five-field technique to deliver 78 Gy to the prostate plus seminal vesicles. Five patients were evaluated. Optimization objectives of both planning approaches concerned dose coverage of the target volumes and the dose distribution in the rectal wall. The two methods were evaluated by comparing dose distributions, the complexity of the resulting plan and the time expenditure to generate and to deliver the plan. RESULTS: For both planning approaches 99% of the target volumes received 95% of the prescribed dose, which complies with our planning objectives. Inverse planning resulted in more conformal dose distributions than forward planning (conformity index: 1.37 versus 1.51). Inverse planning reduced the dose to the rectal wall compared to a manually designed plan, albeit to a small extent. The theoretical probability of severe rectal proctitis and/or stenosis was reduced on average by 1.9% with inverse planning. Maximal sparing of the rectal wall was achieved with inverse planning for a patient whose target volume was partly wrapped around the rectum. The number of segments generated with inverse planning ranged between 33 and 52, and between 9 and 13 segments for manually created segments. CONCLUSION: Dose coverage of the planning target volumes is adequate for both approaches of planning. Inverse planning results in slightly better dose distributions with respect to the rectal wall compared to manual planning, at the cost of an increase of the number of segments by a factor of 3. PMID- 14761029 TI - Quantitative image quality evaluation of pixel-binning in a flat-panel detector for x-ray fluoroscopy. AB - X-ray fluoroscopy places stringent design requirements on new flat-panel (FP) detectors, requiring both low-noise electronics and high data transfer rates. Pixel-binning, wherein data from more that one detector pixel are collected simultaneously, not only lowers the data transfer rate but also increases x-ray counts and pixel signal-to-noise ratio (SNR). In this study, we quantitatively assessed image quality of image sequences from four acquisition methods; no binning and three types of binning; in synthetic images using a clinically relevant task of detecting an extended guidewire in a four-alternative forced choice paradigm. Binning methods were conventional data-line (D) and gate-line (G) binning, and a novel method in which alternate frames in an image sequence used D and G binning. Two detector orientations placed the data lines either parallel or perpendicular to the guide wire. At a low exposure of 0.6 microR (1.548 x 10(-10) C/kg) per frame, irrespective of detector orientation, D binning with its reduced electronic noise was significantly (p<0.1) better than the other acquisition methods. On average, alternate binning performed better than G binning. At a higher exposure of 4.0 microR (10.32 x 10(-10) C/kg) per frame, with data lines parallel to the guidewire, detection with D binning was significantly (p<0.1) better than G binning. However, with data lines perpendicular to the guidewire, G binning was significantly (p<0.1) better than D binning because the partial area effect was reduced. Alternate binning was the best binning method when results were averaged over both orientations, and it was as good as the best binning method at either orientation. In addition, at low and high exposures, alternate binning gave a temporally fused image with a smooth guidewire, an important image quality feature not assessed in a detection experiment. While at high exposure, detection with no binning was as good, or better, than the best binning method, it might be impractical at fluoroscopy imaging rates. A computational observer model based on signal detection theory successfully fit data and was used to predict effects of similar acquisition methods. Results from this study suggest the use of exposure-dependent detector binning in fluoroscopy that switches between D binning and alternate binning at low and high exposures, respectively. PMID- 14761030 TI - Evaluation of the first commercial Monte Carlo dose calculation engine for electron beam treatment planning. AB - The purpose of this study is to perform a clinical evaluation of the first commercial (MDS Nordion, now Nucletron) treatment planning system for electron beams incorporating Monte Carlo dose calculation module. This software implements Kawrakow's VMC++ voxel-based Monte Carlo calculation algorithm. The accuracy of the dose distribution calculations is evaluated by direct comparisons with extensive sets of measured data in homogeneous and heterogeneous phantoms at different source-to-surface distances (SSDs) and gantry angles. We also verify the accuracy of the Monte Carlo module for monitor unit calculations in comparison with independent hand calculations for homogeneous water phantom at two different SSDs. All electron beams in the range 6-20 MeV are from a Siemens KD-2 linear accelerator. We used 10,000 or 50,000 histories/cm2 in our Monte Carlo calculations, which led to about 2.5% and 1% relative standard error of the mean of the calculated dose. The dose calculation time depends on the number of histories, the number of voxels used to map the patient anatomy, the field size, and the beam energy. The typical run time of the Monte Carlo calculations (10,000 histories/cm2) is 1.02 min on a 2.2 GHz Pentium 4 Xeon computer for a 9 MeV beam, 10 x 10 cm2 field size, incident on the phantom 15 x 15 x 10 cm3 consisting of 31 CT slices and voxels size of 3 x 3 x 3 mm3 (total of 486,720 voxels). We find good agreement (discrepancies smaller than 5%) for most of the tested dose distributions. We also find excellent agreement (discrepancies of 2.5% or less) for the monitor unit calculations relative to the independent manual calculations. The accuracy of monitor unit calculations does not depend on the SSD used, which allows the use of one virtual machine for each beam energy for all arbitrary SSDs. In some cases the test results are found to be sensitive to the voxel size applied such that bigger systematic errors (>5%) occur when large voxel sizes interfere with the extensions of heterogeneities or dose gradients because of differences between the experimental and calculated geometries. Therefore, user control over voxelization is important for high accuracy electron dose calculations. PMID- 14761031 TI - Temporal resolving power of perfusion- and BOLD-based event-related functional MRI. AB - Functional magnetic resonance imaging (fMRI) based on both perfusion and blood oxygenation level-dependent (BOLD) contrasts has been widely applied in spatiotemporal mapping of the human brain function. Temporal resolving power of fMRI is limited by the smoothed hemodynamic response function dispersed from the neuronal activity. In this study, temporal modulation transfer functions were utilized to quantify the resolving powers of perfusion and BOLD fMR signals in time domain. The impulse response function was determined using brief visual stimulations and event-related image acquisition schemes. An important feature of arterial spin labeling techniques is that quantitative perfusion and BOLD signals could be simultaneously acquired. This simultaneous BOLD response may arise from signals that are more proximal to capillary beds, and its temporal resolution may be different from that of the typical BOLD response. Therefore, we assessed and compared the temporal resolving capabilities of perfusion, simultaneous BOLD, and the typical BOLD response obtained from the gradient echo EPI pulse sequence. Full-width-at-half-maximums of perfusion and simultaneous BOLD measurements were significantly smaller than that of BOLD ones (4.3+/- 0.6 s vs 5.5 +/- 0.9 s, p<0.02 and 4.7 +/- 1.3 s vs 5.5 +/- 0.9 s, p<0.01, respectively). The corresponding temporal resolving powers of perfusion and simultaneous BOLD signals were statistically better than that of BOLD signals (0.23 +/- 0.03 Hz vs 0.17 +/- 0.02 Hz, p<0.01 and 0.21 +/- 0.04 Hz vs 0.17 +/- 0.02 Hz, p<0.01, respectively). Our results showed that the typical BOLD response was significantly smoothed from the perfusion response, thus resulting in a degraded temporal resolving power. However, results from the simultaneous BOLD and perfusion measurements were not significantly different. Biophysical implications of the experimental outcomes were further investigated using a computer simulation based on the Balloon model. By fitting the measured data into the model, an apparently longer transit time was obtained for the typical BOLD signal (1.7 s), comparing to that for the simultaneous BOLD one (1.2 s). Therefore, the simultaneous BOLD signal was regarded as less susceptible to the variations from local draining veins. Combining the simulation result with the significantly discrepant resolving powers between the two BOLD signals, we speculated that the blurred effects from large vessels played a predominant role that further reduced the temporal resolution of the BOLD-based fMRI from the perfusion response. PMID- 14761032 TI - Verification of IMRT fields by film dosimetry. AB - In intensity modulated radiation therapy (IMRT) the aim of an accurate conformal dose distribution is obtained through a complex process. This ranges from the calculation of the optimal distribution of fluence by the treatment planning system (TPS), to the dose delivery through a multilamellar collimator (MLC), with several segments per beam in the step and shoot approach. The above-mentioned consideration makes mandatory an accurate dosimetric verification of the IM beams. A high resolution and integrating dosimeter, like the radiographic film, permits one to simultaneously measure the dose in a matrix of points, providing a good means of obtaining dose distributions. The intrinsic limitation of film dosimetry is the sensitivity dependence on the field size and on the measurement depth. However, the introduction of a scattered radiation filter permits the use of a single calibration curve for all field sizes and measurement depths. In this paper the quality control procedure developed for dosimetric verification of IMRT technique is reported. In particular a system of film dosimetry for the verification of a 6 MV photon beam has been implemented, with the introduction of the scattered radiation filter in the clinical practice that permits one to achieve an absolute dose determination with a global uncertainty within 3.4% (1 s.d.). The film has been calibrated to be used both in perpendicular and parallel configurations. The work also includes the characterization of the Elekta MLC. Ionimetric independent detectors have been used to check single point doses. The film dosimetry procedure has been applied to compare the measured absolute dose distributions with the ones calculated by the TPS, both for test and clinical plans. The agreement, quantified by the gamma index that seldom reaches the 1.5 value, is satisfying considering that the comparison is performed between absolute doses. PMID- 14761033 TI - Draft 16 of a working document for a proposed standard to be entitled: IEEE standard definitions of terms associated with ferroelectric and related materials. AB - Prior to acceptance of a standard by IEEE, it is important that the document be widely reviewed by experts in the field. This draft version of the standard is being published here to solicit that input. Readers are encouraged to submit any suggestions or corrections to the IEEE ferroelectrics standards committee for inclusion in the final version. PMID- 14761034 TI - Analysis of aging of piezoelectric crystal resonators. AB - Aging of piezoelectric (quartz crystal) resonator has been identified as one of the most important quality control problems of quartz crystal products. Aging is defined as frequency change with time. Aging in quartz resonators can be due to several sources: mass transfer due to contamination inside the resonator enclosure, stress-strain in the resonator blank, quartz defect, etc. In this study, the stress-strain effect, which has been believed as a dominant factor contributing to aging, is studied. The stress-strain effect is caused mainly by the long-term viscoelastic properties of bonding adhesive that attach quartz crystal plate to the ceramic base package. With the available accelerating testing method under elevated temperatures, the stress-strain induced aging in the quartz crystal resonators can be investigated. Because of the miniaturized size of the resonator, a digital image analysis method called image intensity matching technique (IIMT) is applied to obtain deformation patterns in the quartz blank due to thermal load. Our preliminary results showed that the unsymmetric thermal deformations may be a dominant contributing factor to aging. For simulation purposes, finite-element analysis is used to investigate the deformation patterns (i.e., stress-strain distributions) and corresponding natural frequency shift in the piezoelectric resonators. The viscoelastic behavior of mounting adhesives is incorporated into the analysis to show the dominant effect of long-term behavior of stress-strain developed in the crystal resonators. Also, some geometrical aspects-such as uneven mounting supports due to distances, volumes and heights of the adhesives-are simulated in the model. PMID- 14761035 TI - A parametric quartz crystal oscillator. AB - Parametric oscillators have been well studied but currently are not used often. Nevertheless, they could be a low-phase noise solution, at least outside the frequency bandwidth of the resonant circuit. The theoretical aspect of parametric oscillations is briefly reviewed in this paper. Indeed, the basic theory of a simple resistance-inductor-capacitor (RLC) circuit working in parametric conditions easily can be extended toward a resonant loop that includes a quartz crystal resonator. Then, as an application, this study is transposed to a quartz crystal oscillator that has been modeled and tested as a first ptototype. Simulation results are compared with those actually obtained. PMID- 14761036 TI - Cryogenic monolithic sapphire-rutile temperature compensated resonator oscillator. AB - We have tested a new temperature-compensated sapphire resonator as frequency determining element for high-stability microwave oscillator. Temperature compensation has been obtained by coating the sapphire resonator with a thin rutile film. A 2-microm rutile thickness is sufficient to reach turnover temperature higher than 40 K, and a 2 X 10(-12) short-term frequency stability has been obtained. PMID- 14761037 TI - Analysis of a disk-type piezoelectric ultrasonic motor using impedance matrices. AB - The dynamic behavior and the performance characteristics of the disk-type traveling wave piezoelectric ultrasonic motors (USM) are analyzed using impedance matrices. The stator is divided into three coupled subsystems: an inner metal disk, a piezoelectric annular actuator with segmented electrodes, and an outer metal disk with teeth. The effects of both shear deformation and rotary inertia are taken into account in deriving an impedance matrix for the piezoelectric actuator. The impedance matrices for each subsystem then are combined into a global impedance matrix using continuity conditions at the interfaces. A comparison is made between the impedance matrix model and the three-dimensional finite element model of the piezoelectric stator, obtaining the resonance and antiresonance frequencies and the effective electromechanical coupling factors versus circumferential mode numbers. Using the calculated resonance frequency and the vibration modes for the stator and a brush model with the Coulomb friction for the stator and rotor contact, stall torque, and no-load speed versus excitation frequencies are calculated at different preloads. Performance characteristics such as speed-torque curve and the output efficiency of the USM also are estimated using the current impedance matrix and the contact model. The present impedance model can be shown to be very effective in the design of the USM. PMID- 14761038 TI - Force-frequency effect of Y-cut langanite and Y-cut langatate. AB - Most recently, langasite and its isomorphs (LGX) have been advanced as potential substitutes for quartz, owing to their extremely high-quality (Q) factors. At least twice higher Q value of LGX than that of quartz has been reported. High Q translates into potentially greater stability. In order to make such materials practical, the environmental sensitivities must be addressed. One of such sensitivities is the force-frequency effect, which relates the sensitiveness of a resonator to shock and vibration via the third-order (non-Hookean) elastic constants. In this paper, we report measured force-frequency coefficients of a Y cut langanite (LGN) resonator and a Y-cut langatate (LGT) resonator as a function of the azimuthal angle, which is the angle between the crystalline x-axis of a resonator plate and the direction of in-plane diametric force applied to the periphery of the resonator. It was found that the LGN and the LGT behave like AT cut quartz in the polarity of the frequency changes and the existence of zero coefficient angle. The maximum magnitudes of the coefficients of the LGN and the LGT are five and seven times smaller than that of stress-compensated cut (SC-cut) quartz, respectively (or, 7 and 10 times smaller comparing to AT-cut quartz). The coefficients of planar-stress, which represent the superposition of a continuous distribution of periphery stresses, also were obtained as 0.52 X 10(-15) m x s/N and 0.38 X 10(-15) m x s/N for the LGN and the LGT, respectively. PMID- 14761039 TI - Amplitude-frequency effect of Y-cut langanite and langatate. AB - Amplitude-frequency effect of a Y-cut langanite (LGN) resonator and a Y-cut langatate (LGT) resonator were measured. The frequency shifts from the baseline frequency with 1 mA were measured as a function of drive currents up to 28 mA. High-drive current shifted the frequency, but it also heated the crystal locally, causing temperature-related frequency changes. The local heat transfer and its influence on the frequency were analyzed. The amplitude-frequency shift was effectively measured, and was not affected by the temperature-related frequency changes. The 3rd, 5th, and 7th overtones (OT's) were found to behave as soft springs, i.e., resonant frequency decreases as drive current increases. The drive sensitivity coefficients of the 3rd and 5th OT's are in the vicinity of -2 ppb/mA2 for both resonators. The 7th OT's are higher than the other OT's: -5 approximately -7 ppb/mA2. The lowest drive sensitivity is -1.2 ppb/mA2 on the 5th OT of the LGT. PMID- 14761040 TI - Forward planar projection through layered media. AB - A planar forward projection algorithm is combined with ray theory to describe longitudinal propagation through an arbitrary number of randomly oriented isotropic layers. This method first measures the space-time pressure field in a plane, then uses wavevector frequency-domain methods to project the field through layered media and to an arbitrary new plane, not necessarily parallel to the initial plane. The approach is valid for longitudinal propagation through liquid layers and in solids, such as soft tissues, that can be approximated as viscous liquids. The algorithm is verified by propagating the field from a 0.5 MHz planar transducer through a combination of rubber, plastic, and water layers. Hydrophone measurements indicate correlation between measured and simulated fields for angles below the longitudinal critical angles of the layered materials. PMID- 14761041 TI - Volume scattering of distributed microbubbles and its influence on blood flow estimation. AB - In recent years, microbubble contrast agents have become a potential adjunct in Doppler ultrasound diagnosis. In this paper, we show that volume scattering makes the effective band in Doppler spectrum shift downward after injection of microbubbles. Because the insonified volume comprises a collection of distributed microbubbles, the statistical properties such as the autocorrelation function and ensemble average power spectrum of the echoes from a collection of distributed microbubbles were derived first. It can be observed that, beyond a critical frequency, the theoretical volume backscattering cross section derived from the ensemble average power spectrum of microbubbles decreases with frequency. On the contrary, the volume backscattering cross section of red cells increases with frequency. Using two-dimensional (2-D) Fourier transform, the variation in Doppler spectrum caused by different volume backscattering cross section can be demonstrated, and the consequential downward shifts of the estimated Doppler parameters (e.g., the mean and maximum Doppler shifts, and the variance of Doppler power spectrum) after microbubble injection are shown. In addition, it can be observed that the variation gets larger as the transmitted bandwidth increases. And, the variations in Doppler parameters estimated with experimental data are presented to verify the theoretical deviations. PMID- 14761042 TI - Ultrasonic techniques for imaging and measurements in molten aluminum. AB - In order to achieve net shape forming, processing of aluminum (Al) in the molten state is often necessary. However, few sensors and techniques have been reported in the literature due to difficulties associated with molten Al, such as high temperature, corrosiveness, and opaqueness. In this paper, development of ultrasonic techniques for imaging and measurements in molten Al using buffer rods operated at 10 MHz is presented. The probing end of the buffer rod, having a flat surface or an ultrasonic lens, was immersed into molten Al while the other end with an ultrasonic transducer was air-cooled to room temperature. An ultrasonic image of a character "N", engraved on a stainless steel plate immersed in molten Al, and its corrosion have been observed at 780 degrees C using the focused probe in ultrasonic pulse-echo mode. Because cleanliness of molten Al is crucial for part manufacturing and recycling in Al processing, inclusion detection experiments also were carried out using the nonfocused probe in pitch-catch and pulse-echo modes. Backscattered ultrasonic signals from manually added silicon carbide particles, with an average diameter of 50 microm, in molten Al have been successfully observed at 780 degrees C. For optimal image quality, the spatial resolution of the focused probe was crucial, and the high signal-to-noise ratio of the nonfocused probe was the prime factor responsible for the inclusion detection sensitivity using backscattered ultrasonic signals. In addition, it was found that ultrasound could provide an alternative method for evaluating the degree of wetting between a solid material and a molten metal. Our experimental results showed that there was no ultrasonic coupling at the interface between an alumina rod and molten Al up to 1000 degrees C; therefore, no wetting existed at this interface. Also because ultrasonic velocity in alumina is temperature dependent, this rod proved to be able to be used as an in-line temperature monitoring sensor under 1000 degrees C in molten Al. PMID- 14761043 TI - IGBT-based kilovoltage pulsers for ultrasound measurement applications. AB - Two high-voltage pulser designs are presented that offer advantages in some ultrasound measurement applications, such as driving thick ultrasonic source transducers used for broadband measurements of attenuation or hydrophone frequency response and directivity. The pulsers use integrated gate bipolar transistors (IGBTs) as the switching devices, and in one design an output voltage pulse is produced that has a peak amplitude nearly twice that of the supply voltage. The pulsers are inexpensive and relatively easy to construct. The power supply need only provide the average current to charge the capacitors, as opposed to the much higher peak pulse current. With a 1200 V supply and a pulse repetition frequency of 200 Hz, the nondoubling and doubling pulsers provided peak voltages of greater than 1100 V and 2200 V, respectively, into loads ranging from 50 omega to 500 omega. For a 50 omega load, slewing rates of 38 V/ns and 23 V/ns were measured for the nondoubling and doubling pulsers, respectively. For a 500 omega load these values were 56 V/ns and 36 V/ns. PMID- 14761044 TI - A P-matrix-based model for the analysis of SAW transversely coupled resonator filters, including guided modes and a continuum of radiated waves. AB - When designing transversely coupled resonator filters, unexpected spurii are often observed on the high-frequency side of the transfer function. These spurii cannot be described using only the classical waveguide model. Discrete transverse modes inside the grating can be identified if one assumes that the modes have exponential decay outside the grating: however, a continuum of solutions exist in the case of propagating waves outside the grating. A large part of the source excitation may be coupled to these radiated waves. To include this phenomena in the model, a decomposition on the above mentioned continuum was performed. We describe our P-matrix-based model for transversely coupled structures. This model takes into account all guided modes and the continuum. It allows the use of an arbitrary number of acoustical layers and electrical ports. A comparison of the measured and simulated frequency responses is presented for different filters and different metallization thickness showing an excellent agreement. PMID- 14761045 TI - Invariants of electromechanical coupling coefficients in piezoceramics. AB - The relationships between coefficients of electromechanical coupling (CEMC) of various types of piezoceramic resonator (PR) vibrations are considered. Being constant for a given piezoceramic state, the range of variation of piezoceramics dielectric permittivity from a mechanically "free" condition at relatively low frequencies up to an "overall clamped" condition at high frequencies is determined by a consecutive "clamping", caused by a complex of CEMCs of various particular vibrational modes peculiar to the resonator. As the difference between "free" and "overall clamped" permittivities is always determined by the maximal piezomaterial ki3 coupling coefficient, the difference does not depend on the path that was gone through the low-high frequency range, which includes all the vibrational modes possible for a particular PR. The influence of the piezoelectric and elastic anisotropy of lead-zirconate-titanate (PZT) piezoceramic materials on relative CEMC variations was experimentally investigated. PMID- 14761046 TI - An approach for reducing adjacent element crosstalk in ultrasound arrays. AB - A method is presented for active cancellation of crosstalk effects in ultrasonic arrays. The approach makes use of the programmable transmitter waveform generators that are now being used with growing prevalence in diagnostic ultrasound systems. The array's transmit mode transfer function is represented by a transfer function matrix. Elements of this matrix are determined by exciting a single, central element with a wideband waveform and determining the resulting pressure output from the central element and adjacent elements. The desired output then is defined (e.g., finite output from a single, central element) and zero output from all other elements. The transfer function matrix equation can be solved to determine the required excitation functions on both the central array element and its neighbors. These excitation functions will result in reduced evidence of crosstalk on the output signals. Therefore, the single-element, angular-response function is improved. Using superposition, the approach can be extended to beamformed array excitation. A variety of theoretical and experimental results are shown. The method also can be used in the receive mode but with a less satisfactory solution. A transmitting mode experiment based on a prototype five-element transducer has provided results indicating that sidelobes in the angular response can be reduced using this technique. PMID- 14761047 TI - The energy density and power flow of acoustic waves propagating in piezoelectric materials. AB - It is shown that, for plane bulk acoustic wave propagating in arbitrary piezoelectric media, the densities of mechanoelectrical and electromechanical energies are always equal in absolute value and have opposite signs. However, in general, the mechanoelectrical and electromechanical power flows of such wave calculated by traditional expression for Poynting vector do not compensate each other, although the total density of these energies is always equal to zero. Discovered discrepancy based on the dissymmetry of piezoelectric constants with respect to the electrical and mechanical indexes may cause difficulties for calculation of important parameters for practical applications such as energy transport velocity of acoustic waves in piezoelectric materials. PMID- 14761048 TI - The evolution of environmental and genetic sex determination in fluctuating environments. AB - Twenty years ago, Bulmer and Bull suggested that disruptive selection, produced by environmental fluctuations, can result in an evolutionary transition from environmental sex determination (ESD) to genetic sex determination (GSD). We investigated the feasibility of such a process, using mutation-limited adaptive dynamics and individual-based computer simulations. Our model describes the evolution of a reaction norm for sex determination in a metapopulation setting with partial migration and variation in an environmental variable both within and between local patches. The reaction norm represents the probability of becoming a female as a function of environmental state and was modeled as a sigmoid function with two parameters, one giving the location (i.e., the value of the environmental variable for which an individual has equal chance of becoming either sex) and the other giving the slope of the reaction norm for that environment. The slope can be interpreted as being set by the level of developmental noise in morph determination, with less noise giving a steeper slope and a more switchlike reaction norm. We found convergence stable reaction norms with intermediate to large amounts of developmental noise for conditions characterized by low migration rates, small differential competitive advantages between the sexes over environments, and little variation between individual environments within patches compared to variation between patches. We also considered reaction norms with the slope parameter constrained to a high value, corresponding to little developmental noise. For these we found evolutionary branching in the location parameter and a transition from ESD toward GSD, analogous to the original analysis by Bulmer and Bull. Further evolutionary change, including dominance evolution, produced a polymorphism acting as a GSD system with heterogamety. Our results point to the role of developmental noise in the evolution of sex determination. PMID- 14761049 TI - How are deleterious mutations purged? Drift versus nonrandom mating. AB - Accumulation of deleterious mutations has important consequences for the evolution of mating systems and the persistence of small populations. It is well established that consanguineous mating can purge a part of the mutation load and that lethal mutations can also be purged in small populations. However, the efficiency of purging in natural populations, due to either consanguineous mating or to reduced population size, has been questioned. Consequences of consanguineous mating systems and small population size are often equated under "inbreeding" because both increase homozygosity, and selection is though to be more efficient against homozygous deleterious alleles. I show that two processes of purging that I call "purging by drift" and "purging by nonrandom mating" have to be distinguished. Conditions under which the two ways of purging are effective are derived. Nonrandom mating can purge deleterious mutations regardless of their dominance level, whereas only highly recessive mutations can be purged by drift. Both types of purging are limited by population size, and sharp thresholds separate domains where purging is either effective or not. The limitations derived here on the efficiency of purging are compatible with some experimental studies. Implications of these results for conservation and evolution of mating systems are discussed. PMID- 14761050 TI - Contrasting patterns of radiation in African and Australian Restionaceae. AB - The floras of the Mediterranean-climate areas of southern Africa and southwestern Australia are remarkably species rich. Because the two areas are at similar latitudes and in similar positions on their respective continents, they have probably had similar Cenozoic climatic histories. Here we test the prediction that the evolution of the species richness in the two areas followed a similar temporal progression by comparing the rates of lineage accumulation for African and Australian Restionaceae. Restionaceae (Poales) are typical and often dominant elements in the fynbos vegetation of the Cape Floristic Region of southern Africa and the kwongan vegetation of the Southwestern Floristic Province of Western Australia. The phylogeny of the family was estimated from combined datasets for rbcL and trnL-F sequences and a large morphological dataset; these datasets are largely congruent. The monophyly of Restionaceae is supported and a basal division into an African clade (approximately 350 species) and an Australian clade (146 species) corroborated. There is also support for a futher subdivision of these two large sister-clades, but the terminal resolution within the African clade is very weak. Fossil pollen records provided a minimum age of the common ancestor of Australian and African Restionaceae as 64-71 million years ago, and this date was used to calibrate a molecular clock. A molecular clock was rejected by a likelihood ratio test; therefore, rate changes between the lineages were smoothed using nonparametric rate smoothing. The rate-corrected ages were used to construct a plot of lineages through time. During the Palaeogene the Australian lineage diversity increased consistent with the predictions of the constant birthrate model, while the African lineage diversity showed a dramatic increase in diversification rate in the Miocene. Incomplete sampling obscures the patterns in the Neogene, but extending the trends to the modern extant diversity suggests that this acceleration in the speciation rate continued in the African clade, whereas the Australian clade retained a constant diversification rate. The substantial morphological and anatomical similarity between the African and Australian Restionaceae appear to preclude morphological innovations as possible explanations for the intercontinental differences. Most likely these differences are due to the greater geographical extent and ecological variation in temperate Australia than temperate Africa, which might have provided refugia for basal Restionaceae lineages, whereas the more mountainous terrain of southern Africa might have provided the selective regimes for a more rapid, recent speciation. PMID- 14761051 TI - A multilocus genealogical approach to phylogenetic species recognition in the model eukaryote Neurospora. AB - To critically examine the relationship between species recognized by phylogenetic and reproductive compatibility criteria, we applied phylogenetic species recognition (PSR) to the fungus in which biological species recognition (BSR) has been most comprehensively applied, the well-studied genus Neurospora. Four independent anonymous nuclear loci were characterized and sequenced from 147 individuals that were representative of all described outbreeding species of Neurospora. We developed a consensus-tree approach that identified monophyletic genealogical groups that were concordantly supported by the majority of the loci, or were well supported by at least one locus but not contradicted by any other locus. We recognized a total of eight phylogenetic species, five of which corresponded with the five traditional biological species, and three of which were newly discovered. Not only were phylogenetic criteria superior to traditional reproductive compatibility criteria in revealing the full species diversity of Neurospora, but also significant phylogenetic subdivisions were detected within some species. Despite previous suggestions of hybridization between N. crassa and N. intermedia in nature, and the fact that several putative hybrid individuals were included in this study, no molecular evidence in support of recent interspecific gene flow or the existence of true hybrids was observed. The sequence data from the four loci were combined and used to clarify how the species discovered by PSR were related. Although species-level clades were strongly supported, the phylogenetic relationships among species remained difficult to resolve, perhaps due to conflicting signals resulting from differential lineage sorting. PMID- 14761052 TI - Reproductive isolation and phylogenetic divergence in Neurospora: comparing methods of species recognition in a model eukaryote. AB - We critically examined methods for recognizing species in the model filamentous fungal genus Neurospora by comparing traditional biological species recognition (BSR) with more comprehensive applications of both BSR and phylogenetic species recognition (PSR). Comprehensive BSR was applied to a set of 73 individuals by performing extensive crossing experiments and delineating biological species based on patterns of reproductive success. Within what were originally considered two species, N. crassa and N. intermedia, we recognized four reproductively isolated biological species. In a concurrent study (Dettman et al. 2003), we used genealogical concordance of four independent nuclear loci to recognize phylogenetic species in Neurospora. Overall, the groups of individuals identified as species were similar whether recognized by reproductive success or by phylogenetic criteria, and increased genetic distance between parents was associated with decreased reproductive success of crosses, suggesting that PSR using genealogical concordance can be used to reliably recognize species in organisms that are not candidates for BSR. In one case, two phylogenetic species were recognized as a single biological species, indicating that significant phylogenetic divergence preceded the development of reproductive isolation. However, multiple biological species were never recognized as a single phylogenetic species. Each of the putative N. crassa x N. intermedia hybrids included in this study was confidently assigned to a single species, using both PSR and BSR. As such, no evidence for a history of hybridization in nature among Neurospora species was observed. By performing reciprocal mating tests, we found that mating type, parental role, and species identity of parental individuals could all influence the reproductive success of matings. We also observed sympatry-associated sexual dysfunction in interspecific crosses, which was consistent with the existence of reinforcement mechanisms. PMID- 14761053 TI - Pollen transfer by hummingbirds and bumblebees, and the divergence of pollination modes in Penstemon. AB - We compared pollen removal and deposition by hummingbirds and bumblebees visiting bird-syndrome Penstemon barbatus and bee-syndrome P. strictus flowers. One model for evolutionary shifts from bee pollination to bird pollination has assumed that, mostly due to grooming, pollen on bee bodies quickly becomes unavailable for transfer to stigmas, whereas pollen on hummingbirds has greater carryover. Comparing bumblebees and hummingbirds seeking nectar in P. strictus, we confirmed that bees had a steeper pollen carryover curve than birds but, surprisingly, bees and birds removed similar amounts of pollen and had similar per-visit pollen transfer efficiencies. Comparing P. barbatus and P. strictus visited by hummingbirds, the bird-syndrome flowers had more pollen removed, more pollen deposited, and a higher transfer efficiency than the bee-syndrome flowers. In addition, P. barbatus flowers have evolved such that their anthers and stigmas would not easily come into contact with bumblebees if they were to forage on them. We discuss the role that differences in pollination efficiency between bees and hummingbirds may have played in the repeated evolution of hummingbird pollination in Penstemon. PMID- 14761054 TI - Tracking a genetic signal of extinction-recolonization events in a neotropical tree species: Vouacapoua americana Aublet in French Guiana. AB - Drier periods from the late Pleistocene and early Holocene have been hypothesized to have caused the disappearance of various rainforest species over large geographical areas in South America and restricted the extant populations to mesic sites. Subsequent improvement in climatic conditions has been associated with recolonization. Changes in population size associated with these extinction recolonization events should have affected genetic diversity within species. However, these historical hypotheses and their genetic consequences have rarely been tested in South America. Here, we examine the diversity of the chloroplast and nuclear genomes in a Neotropical rainforest tree species, Vouacapoua americana (Leguminosae, Caesalpinioideae) in French Guiana. The chloroplast diversity was analyzed using a polymerase chain reaction-restriction fragment length polymorphism method (six pairs of primers) in 29 populations distributed over most of French Guiana, and a subset of 17 populations was also analyzed at nine polymorphic microsatellite loci. To determine whether this species has experienced extinction-recolonization, we sampled populations in areas supposedly not or only slightly affected by climatic changes, where the populations would not have experienced frequent extinction, and in areas that appear to have been recently recolonized. In the putatively recolonized areas, we found patches of several thousands of hectares homogeneous for chloroplast variation that can be interpreted as the effect of recolonization processes from several geographical origins. In addition, we observed that, for both chloroplast and nuclear genomes, the populations in newly recolonized areas exhibited a significantly smaller allelic richness than others. Controlling for geographic distance, we also detected a significant correlation between chloroplast and nuclear population differentiation. This result indicates a cytonuclear disequilibrium that can be interpreted as a historical signal of a genetic divergence between fragmented populations. In conclusion, the spatial genetic structure of contemporary V. americana populations shows evidence that this species has experienced large extinction-recolonization events, which were possibly caused by past climatic change. PMID- 14761055 TI - Costs and benefits of evolving under experimentally enforced polyandry or monogamy. AB - Reproduction has classically been viewed as a predominantly cooperative process. However, over the last 20 years this concept has steadily yielded ground to one of continual conflict in which the interests of the sexes are typically discordant. Within this framework, males and females are seen to be locked into a perpetual arms race, each adaptation by one sex promoting the evolution of countermeasures in the other sex. However, under strict genetic monogamy, the interests of the sexes become congruent, and hence antagonistic coevolution does not occur. We subjected the fly Sepsis cynipsea, a species with conspicuous sexual conflict, to experimentally enforced monogamy or polyandry for 29 generations and evaluated the microevolutionary consequences. We found that there were longevity costs to females consistent with sexually antagonistic coevolution. However, our measure of female fitness, offspring emergence, did not differ between treatments, even though life-history characters such as fertility and fecundity did. Results are discussed in terms of costs and benefits of sexual selection and sexual conflict. PMID- 14761056 TI - Allometric and nonallometric components of Drosophila wing shape respond differently to developmental temperature. AB - Phenotypic plasticity of wing size and shape of Drosophila simulans was analyzed across the entire range of viable developmental temperatures with Procrustes geometric morphometric method. In agreement with previous studies, size clearly decreases when temperature increases. Wing shape variation was decomposed into its allometric (24%) and nonallometric (76%) components, and both were shown to involve landmarks located throughout the entire wing blade. The allometric component basically revealed a progressive, monotonous variation along the temperature. Surprisingly, nonallometric shape changes were highly similar at both extremes of the thermal range, suggesting that stress, rather than temperature per se, is the key developmental factor affecting wing shape. PMID- 14761057 TI - Population genetics of accessory gland proteins and sexual behavior in Drosophila melanogaster populations from Evolution Canyon. AB - Evolution Canyon in Lower Nahal Oren, Mount Carmel, Israel has been identified as a location promoting sympatric speciation. Several previous studies on Drosophila melanogaster populations from the two disparate slopes of the canyon suggest that these two populations are experiencing incipient speciation. However, recent microsatellite data did not reveal the expected level of population differentiation. Given the importance of this system for studying speciation, we set out to test two predictions of the incipient speciation hypothesis--genetic differentiation and sexual isolation. We sequenced six different Acp genes from isofemale lines from the south-facing slope (11 lines) and north-facing slope (nine lines) of Evolution Canyon. We found no evidence of genetic differentiation between the two slopes (F(ST) = -0.03). We also conducted mate choice tests, using intraslope F1 hybrids between different isofemale lines. There was no significant departure from random mating in mixtures of flies from the two slopes. Our results provide further indication that it is unlikely these two populations are experiencing incipient speciation. We discuss our results in light of the discrepancies that have been published on this enigmatic D. melanogaster system from Evolution Canyon. PMID- 14761058 TI - Mosquito mortality and the evolution of malaria virulence. AB - Several laboratory studies of malaria parasites (Plasmodium sp.) and some field observations suggest that parasite virulence, defined as the harm a parasite causes to its vertebrate host, is positively correlated with transmission. Given this advantage, what limits the continual evolution of higher parasite virulence? One possibility is that while more virulent strains are more infectious, they are also more lethal to mosquitoes. In this study, we tested whether the virulence of the rodent malaria parasite P. chabaudi in the laboratory mouse was correlated with the fitness of mosquitoes it subsequently infected. Mice were infected with one of seven genetically distinct clones of P. chabaudi that differ in virulence. Weight loss and anemia in infected mice were monitored for 16-17 days before Anopheles stephensi mosquitoes were allowed to take a blood meal from them. Infection virulence in mice was positively correlated with transmission to mosquitoes (infection rate) and weakly associated with parasite burden (number of oocysts). Mosquito survival fell with increasing oocyst burden, but there was no overall statistically significant relationship between virulence in mice and mosquito mortality. Thus, there was no evidence that more virulent strains are more lethal to mosquitoes. Both vector survival and fecundity depended on parasite clone, and contrary to expectations, mosquitoes fed on infections more virulent to mice were more fecund. The strong parasite genetic effects associated with both fecundity and survival suggests that vector fitness could be an important selective agent shaping malaria population genetics and the evolution of phenotypes such as virulence in the vector. PMID- 14761059 TI - Hierarchical analysis of colony and population genetic structure of the eastern subterranean termite, Reticulitermes flavipes, using two classes of molecular markers. AB - Termites (Isoptera) comprise a large and important group of eusocial insects, yet, in contrast to the eusocial Hymenoptera (ants, bees, wasps), the breeding systems of termites remain poorly understood. In this study, I inferred the breeding system of the subterranean termite Reticulitermes flavipes based on colony and population genetic structure as determined from microsatellite and mitochondrial DNA markers. Termites were sampled from natural wood debris from three undisturbed, forested sites in central North Carolina. In each site, two transects separated by 1 km were sampled at approximately 15-m intervals. A total of 1272 workers collected from 57 collection points were genotyped at six microsatellite loci, and mitochondrial DNA haplotype was determined for a subset of these individuals using either restriction fragment length polymorphism or sequence variation in the AT-rich region. Colonies appeared to be localized: workers from the 57 collection points represented 56 genetically distinct colonies with only a single colony occupying two collection points located 15 m apart. Genetic analysis of family structure and comparisons of estimates of F statistics (F(IT), F(IC), F(CT)) and coefficients of relatedness (r) among nestmate workers with results of computer simulations of potential breeding systems suggested that 77% of all colonies were simple families headed by outbred monogamous pairs, whereas the remaining colonies were extended (inbred) families headed by low numbers of neotenics (about two females and one male) who were the direct offspring of the colony founders. There was no detectable isolation by distance among colonies along transects, suggesting that colony reproduction by budding is not common and that dispersal of reproductives during mating flights is not limited over this distance. Higher-level analysis of the microsatellite loci indicated weak but significant differentiation among sites (F(ST) = 0.06), a distance of 16-38 km, and between transects within sites (F(ST) = 0.06), a distance of 1 km. No significant differentiation at either the transect or site level was detected in the mitochondrial DNA sequence data. These results indicate that the study populations of R. flavipes have a breeding system characterized by monogamous pairs of outbred reproductives and relatively low levels of inbreeding because most colonies do not live long enough to produce neotenics, and those colonies that do generate neotenics contain an effectively small number of them. PMID- 14761060 TI - Evolution of subterranean diving beetles (Coleoptera: Dytiscidae: Hydroporini, Bidessini) in the arid zone of Australia. AB - Calcrete aquifers in arid inland Australia have recently been found to contain the world's most diverse assemblage of subterranean diving beetles (Coleoptera: Dytiscidae). In this study we test whether the adaptive shift hypothesis (ASH) or the climatic relict hypothesis (CRH) is the most likely mode of evolution for the Australian subterranean diving beetles by using a phylogeny based on two sequenced fragments of mitochondrial genes (CO1 and 16S-tRNA-ND1) and linearized using a relaxed molecular clock method. Most individual calcrete aquifers contain an assemblage of diving beetle species of distantly related lineages and/or a single pair of sister species that significantly differ in size and morphology. Evolutionary transitions from surface to subterranean life took place in a relatively small time frame between nine and four million years ago. Most of the variation in divergence times of the sympatric sister species is explained by the variation in latitude of the localities, which correlates with the onset of aridity from the north to the south and with an aridity maximum in the Early Pliocene (five mya). We conclude that individual calcrete aquifers were colonized by several distantly related diving beetle lineages. Several lines of evidence from molecular clock analyses support the CRH, indicating that all evolutionary transitions took place during the Late Miocene and Early Pliocene as a result of aridification. PMID- 14761061 TI - Initial stages of reproductive isolation in two species of the endangered Sonoran topminnow. AB - Long-term geographic isolation can result in reproductive incompatibilities due to forces such as mutation, genetic drift, and differential selection. In the Sonoran topminnow, molecular genetic studies of mtDNA, microsatellites, and MHC genes have shown that the endangered Gila and Yaqui topminnows are substantially different, suggesting that divergence took place approximately two million years ago. Here we examined hybrid crosses and backcrosses between these two allopatric taxa to evaluate the accumulation of postmating barriers to reproduction. These results are then compared with results from a previous study where male topminnows were shown to mate assortatively with conspecific females. Despite their preference for conspecific mates, both types of interspecific crosses successfully produced offspring. There was evidence of reduced hybrid fitness, including smaller mean brood size and male-biased sex ratio, for some classes of backcrosses. Brood sizes and interbrood intervals varied significantly when hybrids were subdivided into different cross categories. Our results illustrate the importance of distinctly defining hybrid classes in studies of reproductive isolation. To our knowledge, this is the first such detailed evolutionary analysis in endangered fish taxa. PMID- 14761062 TI - A test of alternative hypotheses for the evolution of reproductive isolation between spadefoot toads: support for the reinforcement hypothesis. AB - How do species that interbreed become reproductively isolated? If hybrids are less fit than parental types, natural selection should promote reproductive isolation by favoring the evolution of premating mechanisms that prevent hybridization (a process termed reinforcement). Although reinforcement should generate a decline in hybridization over time, countervailing forces of gene flow and recombination are thought to preclude natural selection from enhancing and finalizing reproductive isolation. Here, I present recent estimates of hybridization frequency between two species of spadefoot toad, Spea multiplicata and S. bombifrons. I compare these recent measures of hybrid frequency with previously published estimates and show that hybridization between these species has declined precipitously over the past 27 years. Although previous studies suggest that reinforcement possibly accounts for this decline in hybrids over time, three alternative hypotheses also can explain the observed decrease in hybridization. First, if one of the two interacting species becomes rare, opportunities for and incidence of hybridization may decrease. Second, if one of the two interacting species is initially rare, hybridization may be initially common if the rare species has difficulty locating conspecific mates. Third, if hybrids are produced only in particular environments, hybrid frequency may decline if habitat changes result in loss of those environments that promote hybrid formation. I found no support for these three alternative explanations of the decline in hybrids. Instead, reinforcement appears to best account for the evolution of enhanced reproductive isolation between these species. Moreover, the finding that hybridization declined precipitously in only 27 years suggests that many systems that have undergone reinforcement may be overlooked because reproductive isolation between the interacting populations or species may already be complete. PMID- 14761063 TI - Reconciling actual and inferred population histories in the house finch (Carpodacus mexicanus) by AFLP analysis. AB - The house finch (Carpodacus mexicanus) is a native songbird of western North America that was introduced to the eastern United States and Hawaiian Islands in historic times. As such, it provides an unusually good opportunity to test the ability of molecular markers to recover recent details of a known population history. To investigate this prospect, genetic variation in 172 individuals from 16 populations in the western and eastern United States, southeastern Canada, Hawaiian Islands, and Mexico, as well as genetic variation in the closely related purple finch (Carpodacus purpureus) and Cassin's finch (Carpodacus cassinii) was studied by a semi-automated fluorescence-labeled amplified fragment length polymorphism (AFLP) marker system. A total of 363 markers were generated, of which 258 (71.2%) were polymorphic among species, 166 (61.4%) polymorphic among house finch subspecies, and 157 (60.2%) polymorphic among populations within the frontalis subspecies complex. Heterozygosities and interpopulation divergences revealed by the analysis appeared relatively low at all taxonomic levels, but there are few similar studies in avian populations with which to compare results. Whereas the known population history predicts that both eastern and Hawaiian finches should have been derived from within western populations, tree analysis using both populations and individuals as units suggests weak monophyly of eastern populations and indicates that Hawaiian populations are not clearly derived from California populations. However, the genetic distinctiveness of native and recently founded populations was disclosed by analyses of molecular variance as well as by a model-based assignment approach in which 98%, 94%, and 99% individuals from western, Hawaiian, and eastern regions, respectively, were assigned correctly to their populations without using prior information on population of origin, suggesting that these recent introductions have resulted in detectable differentiation without substantial loss of AFLP diversity. Our results indicate that AFLPs are a useful tool for population genetic and evolutionary studies of birds, particularly as a prelude to finding molecular markers linked to traits subjected to recent adaptive evolution. PMID- 14761064 TI - Genetic differentiation at nuclear and mitochondrial loci among large white headed gulls: sex-biased interspecific gene flow? AB - We measured genetic differentiation among species of large white-headed gulls using mitochondrial (cytochrome b haplotypes) and nuclear (microsatellites) markers. Additional information was added using a previously published study of allozymes on the same species. Levels of differentiation among species at nuclear markers are much lower than would be expected for avian species and are not concordant with the level of differentiation in mitochondrial markers. This discrepancy is best explained by a combination of recent species origin and interspecific gene flow after speciation. The data also suggest that female mediated gene flow is reduced compared to male-mediated gene flow, either due to behavioral bias or due to stronger counterselection of female hybrids in accordance with Haldane's rule for ZW species. Whatever the reasons for the low differentiation of the species' nuclear gene pools, the extensive similarity of their nuclear genome demonstrates that selection on a limited number of characters is an important factor in establishing and maintaining clear-cut phenotypic differences between these species and suggests that the number of loci involved in this process is quite low. This situation may not be exceptional in birds, indeed a number of studies have found similarly low level of differentiation in nuclear markers among congeneric bird species, although usually based on a single set of markers. Because hybridization is a widespread phenomenon in birds, many of these cases might be due to interspecific gene flow. PMID- 14761065 TI - Estimating selection on neonatal traits in red deer using elasticity path analysis. AB - Van Tienderen recently published a method that links selection gradients between a phenotypic trait and multiple fitness components with the effects of these fitness components on the population growth rate (mean absolute fitness). The method allows selection to be simultaneously estimated across multiple fitness components in a population dynamic framework. In this paper we apply the method to a population of red deer living in the North Block of the Isle of Rum, Scotland. We show that (1) selection on birth date and birth weight can operate through multiple fitness components simultaneously; (2) our estimates of the response to selection are consistent with the observed change in trait values that we cannot explain with environmental and phenotypic covariates; (3) selection on both traits has fluctuated over the course of the study; (4) selection operates through different fitness components in different years; and (5) no environmental covariates correlate with selection because different fitness components respond to density and climatic variation in contrasting ways. PMID- 14761067 TI - Information about transmission opportunities triggers a life-history switch in a parasite. AB - Many microbial pathogens can switch to new hosts or adopt alternative transmission routes as environmental conditions change, displaying unexpected flexibility in their infection pathways and often causing emerging diseases. In contrast, parasitic worms that must develop through a fixed series of host species appear less likely to show phenotypic plasticity in their transmission pathways. Here, I demonstrate experimentally that a trematode parasite, Coitocaecum parvum, can accelerate its development and rapidly reach precocious maturity in its crustacean intermediate host in the absence of chemical cues emanating from its fish definitive host. Juvenile trematodes can also mature precociously when the mortality rate of their intermediate hosts is increased. Eggs produced by precocious adults hatch into viable larvae, capable of pursuing the parasite's life cycle. In the absence of chemical cues from fish hosts, the size of eggs released by precocious trematodes in their intermediate hosts becomes more variable, possibly indicating a bet-hedging strategy. These results illustrate that parasitic worms with complex life cycles have development and transmission strategies that are more plastic than commonly believed, allowing them to skip one host in their cycle when they perceive limited opportunities for transmission. PMID- 14761066 TI - Energy and the rate of evolution: inferences from plant rDNA substitution rates in the western Pacific. AB - In this study, we compare rDNA substitution rates for a group of closely related plant species in the western Pacific that exist in different biomes. The results of this comparison indicate higher rates of substitution for species living in habitats with greater biologically available energy. We interpret that finding as potentially important in understanding evolution because of its implication that substitution rate may be a function of biologically available energy and its correlate, productivity. The relevance of this research is twofold. First, contrasting closely related species across different biomes allows for a comparison between rates of molecular evolution across different energetic/productivity regimes while controlling for phylogenetically influenced variation. Second, the research indicates some of the design parameters for future studies that are required to explore the importance of this relationship among different groups of related organisms. If higher rates of molecular evolution where there is greater available energy are found to be widespread this might bring an additional dimension to the understanding of macroevolutionary pattern and process. PMID- 14761068 TI - Correlated evolution of conspicuous coloration and body size in poison frogs (Dendrobatidae). AB - Conspicuous coloration is often used in combination with chemical defenses to deter predators from attacking. Experimental studies have shown that the avoidance inducing effect of conspicuous prey coloration increases with increasing size of pattern elements and with increasing body size. Here we use a comparative approach to test the prediction from these findings, namely that conspicuous coloration will evolve in tandem with body size. In our analysis, we use a previously published mitochondrial DNA-based phylogeny and comparative analysis of independent contrasts to examine if evolutionary shifts in color pattern have been associated with evolutionary changes in body size in aposematic poison frogs (Anura: Dendrobatidae). Information on body size (snout to vent length) and coloration were obtained from the literature. Two different measures of conspicuousness were used, one based on rankings by human observers and the other based on computer analysis of digitized photographs. The results from comparative analyses using either measure of coloration indicated that avoidance inducing coloration and body size have evolved in concert in poison frogs. Results from reconstruction of character change further indicate that the correlated evolution of size and coloration has involved changes in both directions within each of the different clades of the phylogenetic tree. This finding is consistent with the hypothesis that selection imposed by visually guided predators has promoted the evolution of larger body size in species with conspicuous coloration, or enhanced evolution of conspicuous coloration in larger species. PMID- 14761069 TI - Inbreeding and interbreeding in Darwin's finches. AB - Studies of inbreeding and interspecific hybridization are generally pursued separately with different metrics. There is a need to integrate them because they have the common goal of seeking an understanding of the genetic and ecological basis of fitness variation in populations. We use mean expected heterozygosity as an axis of variation on which to compare the fitness of inbreeding and hybridizing Darwin's finches (Geospiza scandens and G. fortis) relative to the fitness of matched outbred controls. We find that relative fitness of inbred finches is less than one in the 1991 cohorts of both species. Inbreeding depression is stronger in the species (G. scandens) with the lower genetically effective population size. Relative fitness of hybrids (backcrosses) in the same cohort of G. scandens is greater than one. Evidence of heterosis in G. fortis is mixed. Thus the two interbreeding species displayed somewhat different fitness patterns under the same set of environmental conditions. Hybridization may enhance fitness to different degrees by counteracting the effects of inbreeding depression, by other additive and nonadditive genetic effects, and by producing phenotypes well suited to exploit particular ecological conditions. PMID- 14761070 TI - Management of hypertension--more focus on treatment targets. PMID- 14761071 TI - Left ventricular hypertrophy regression with antihypertensive treatment: focus on Candesartan. PMID- 14761072 TI - Doxazosin GITS versus hydrochlorothiazide as add-on therapy in patients with uncontrolled hypertension. AB - The objective of this prospective, randomized, open-label, parallel-arm comparative study, with a 4-month follow-up, was to assess the antihypertensive efficacy, tolerability and metabolic safety of doxazosin GITS (gastrointestinal therapeutic system) 4-8 mg/day vs hydrochlorothiazide (HCTZ) 12.5-25 mg/day as add-on therapy in patients not controlled with monotherapy with other drugs. Ninety-eight patients completed the study (mean age 57.4 +/- 15 years, 53% female). Mean systolic/diastolic blood pressure reduction was 8.2/4.5 mmHg in the HCTZ group and 8.9/5.0 mmHg in the doxazosin GITS group, and a strict blood pressure control was achieved in 79% and 83% of the patients, respectively. The incidence rates of adverse events were low and similar in both groups. However, metabolic differences were seen between the groups, doxazosin GITS vs HCTZ, respectively: total cholesterol (mg/dl) 210 +/- 53 vs 231 +/- 62 (p < 0.05), low density lipoprotein (LDL) cholesterol (mg/dl) 139 +/- 40 vs 161 +/- 57 (p < 0.01), high-density lipoprotein (HDL) cholesterol (mg/dl) 58 +/- 16 vs 48 +/- 13 (p < 0.01), HDL/total cholesterol ratio 27.6 +/- 8 vs 21.2 +/- 7 (p < 0.001), plasma uric acid (mg/dl) 5.3 +/- 2.6 vs 6.8 +/- 3.1 (p < 0.05) and serum potassium (mEq/l) 4.1 +/- 1.3 vs. 3.7 +/- 1.2 (p < 0.01). In conclusion, doxazosin GITS has a tolerability and efficacy profile similar to low doses of thiazide diuretics, with a better evolution of metabolic and electrolyte parameters. Therefore, in patients not controlled with monotherapy, doxazosin GITS can be considered an alternative to the addition of thiazide diuretics. PMID- 14761073 TI - The effects of replacing dihydropyridine calcium-channel blockers with angiotensin II receptor blocker on the quality of life of hypertensive patients. AB - Hypertension is a major risk factor for cardiovascular events and the goal of treating hypertension is to prevent complications due to these events. However, some other properties, including few side-effects and improvement of the quality of life (QOL), are desirable in a drug as well as its antihypertensive effect. Dehydropydine calcium-channel blockers (DCCBs) are the most frequently used antihypertensive agents in Japan. The antihypertensive effect of DCCBs is satisfactory, but side-effects, e.g. nocturia, flushing and palpitations, are a problem. The aim was to evaluate the effects of a change of treatment from DCCBs on the QOL of hypertensive patients. An open study was performed to evaluate the effects of switching treatment from DCCBs to angiotensin II receptor blocker (ARB) therapy on the QOL of hypertensive patients. The ARBs have been reported to be effective and well-tolerated antihypertensive drugs. Candesartan cilexetil was selected because it is the most frequently used ARB in Japan. One hundred patients with mild to moderate hypertension, being treated with DCCBs, were randomly selected to receive candesartan cilexetil (8-12 mg once a day). The patients were followed for 3 months, while blood pressure (BP), side-effects and QOL were monitored. BP was equally well controlled before and after the change of antihypertensive therapy. The candesartan cilexetil-treated patients exhibited improvement of several aspects of QOL, including general symptoms, physical symptoms and well-being, work and satisfaction and sleep scale. Emotional state and cognitive function also improved. Patients aged 65 years or younger achieved significant improvement of sexual function. Changing treatment from DCCBs to ARB therapy achieved equal BP control with a lower drug dose. Moreover, the change to cadesartan cilexetil had a positive impact on the QOL. PMID- 14761074 TI - Effect of the angiotensin II antagonist valsartan on sexual function in hypertensive men. AB - The present open and prospective study was performed to investigate the effect of the angiotensin II receptor blocker (ARB) valsartan on sexual function in hypertensive males. Patients who were either newly treated or who were switched from other treatment regimens received valsartan 80-160 mg/day. Blood pressure decreased from an average of 158/94 mmHg to 136/82 mmHg during the 6 months of treatment (p < 0.001). The patients' sexual function was assessed before valsartan and after 6 months of treatment using the International Index of Erectile Function (IIEF), an internationally validated 15-item questionnaire. The IIEF addresses the relevant domains of male sexual function, i.e. erectile function, orgasmic function, sexual desire, intercourse satisfaction and overall satisfaction. At baseline, 75.4% of the total group of 3502 patients investigated and 65.0% of the subgroup of patients without previous antihypertensive treatment (n = 952) could be diagnosed as having erectile dysfunction (ED) according to the IIEF. Valsartan therapy markedly reduced ED in these groups to 53% and 45% (p < 0.0001), respectively. Improved ED was associated with highly significant improvements in orgasmic function, intercourse and overall satisfaction both in the total and previously untreated groups. In addition, sexual desire averaged 5.64 +/- 1.99 IIEF units in the total and 5.99 +/- 2.03 in the group without antihypertensive treatment at baseline. Valsartan markedly increased these numbers to 6.82 +/- 1.72 and 7.06 +/- 1.68 (p < 0.0001), respectively. The results of our open study suggest that the ARB valsartan improves sexual function in hypertensive males. PMID- 14761075 TI - Efficacy and tolerability profile of nebivolol vs atenolol in mild-to-moderate essential hypertension: results of a double-blind randomized multicentre trial. AB - The objective of this 12-week double-blind randomized multicentre study was to compare the efficacy and tolerability of nebivolol, a recently developed beta blocking agent with vasodilating properties, to the classical beta-blocker atenolol. After a placebo run-in phase, 205 mild-to-moderate middle-age essential hypertensives were randomized to either nebivolol 5 mg daily (n = 105) or atenolol 100 mg daily (n = 100) over a period of 12 weeks. The primary endpoint of the study was the change in sitting systolic and diastolic blood pressure (SBP and DBP respectively) from baseline to week 12 of treatment. The two drugs induced similar significant antihypertensive effects, the SBP and DBP reduction amounting to -18.2 +/-14.0 and -14.6 +/-7.9 mmHg (mean +/- SD) for atenolol and 19.1 +/-12.9 and -14.8 +/- 7.1 for nebivolol (p < 0.01 for all). This was the case also for standing blood pressure. Sitting and standing heart rate values were significantly reduced by both drugs, the bradicardic response induced by nebivolol treatment being significantly less than atenolol. Distribution of responders and non- responders was similar for nebivolol and atenolol, while the former drug showed a better tolerability profile and a lower incidence of side effects. These data provide evidence, that, for the same antihypertensive effects, nebivolol shows a better tolerability profile than atenolol and a lower incidence of adverse effects. PMID- 14761076 TI - Efficacy and tolerability of the combination valsartan/hydrochlorothiazide compared with amlodipine in a mild-to-moderately hypertensive Brazilian population. AB - Most hypertensive patients need more than one drug to reach recommended blood pressure targets. We investigated the effects on 24-h ambulatory blood pressure (ABP) of the angiotensin-receptor blocker, valsartan, in combination with hydrochlorothiazide (HCTZ), compared with the calcium-channel blocker amlodipine in a Brazilian population in a multicentre, double-blind, double-dummy, parallel group, controlled study in 373 patients with essential hypertension. After a 2 week washout period, patients with a mean sitting systolic blood pressure (SBP) of 160-190 mmHg were randomized to receive either valsartan 160 mg o.d., or amlodipine 5 mg o.d. for 2 weeks and subsequently force-titrated to valsartan 160 mg/HCTZ 25 mg o.d. or amlodipine 10 mg o.d. This regimen was continued until the end of the study at week 8. The primary efficacy parameter was the change from baseline to week 8 in mean 24-h SBP. Secondary endpoints were change in mean 24-h diastolic blood pressure (DBP), tolerability and safety of treatments. Valsartan/HCTZ achieved a mean reduction in systolic ABP of -19.1 +/- 11.3 mmHg compared with -20.7 +/- 12.0 mmHg with amlodipine (p = 0.324 for the comparison) and in diastolic ABP by -11.1 +/- 7.4 mmHg vs -11.6 +/- 7.2 mmHg by amlodipine (p = 0.853 for the comparison). The valsartan/HCTZ group exhibited markedly lower rates of adverse events and discontinuations than the amlodipine group. Peripheral oedemas were far more frequent with amlodipine than with valsartan/ HCTZ (1.6% with valsartan/HCTZ; 16.8% with amlodipine). Thus, the valsartan 160 mg/HCTZ 25 mg combination appears to be as efficacious as amlodipine 10 mg in this patient population but better tolerated. PMID- 14761077 TI - The leading object. PMID- 14761078 TI - Challenges of accurately defining the nutrient requirements of heat-stressed poultry. AB - The number of factors that influence the nutrient requirements of heat-stressed poultry, through alterations in metabolism and biological responses, are much greater than those that influence the nutrient requirements of poultry at thermoneutral temperatures. Therefore, multiple dietary nutrient specifications adjusted for different intensities of heat stress experienced in different geographical locations, and for different management options used to combat such conditions, are likely to be required by poultry nutritionists. Financial constraints on the construction of suitable research facilities have limited the accumulation of knowledge concerning dietary nutrient specifications for heat stressed poultry. Thus, input from commercial enterprises has contributed significantly to the accumulation of existing knowledge. Knowledge of the energy and amino acid requirements of birds during heat stress are best known, whereas vitamins and trace minerals remain among the least-researched groups of nutrients. PMID- 14761079 TI - Effects of bird age, density, and molt on behavioral profiles of two commercial layer strains in cages. AB - Two commercial strains, Hy-Line W-36 and DeKalb XL, were moved to a laying house at 18 wk of age. They were housed 6 hens/layer cage at 2 densities (361 and 482 cm2/bird) with 2 replications each per strain/density combination. The high density treatment contained 24 hens/replication and the low-density treatment contained 18 hens/replication for a total of 168 hens. Production parameters were measured during the first egg production cycle, the molt period, and the first 4 wk of the second lay cycle (20 to 68 wk of age). Behavioral observations were taken during 2 consecutive d at 26, 34, 43, 51, 62, 64, and 68 wk of age to examine behavioral patterns. Modified Hansen's tests were conducted concurrently to provide indication of the fearfulness levels of hens at the various stages of production. The production characteristics were similar for both strains. The hens kept at the higher density had lower (P < 0.01) hen-day production and (P < 0.05) daily egg mass. Appetitive behaviors were not affected by strain or density but were affected by the age of the hen and by molting. During the molt, feeding and drinking behavioral acts were fewer (P < 0.05) at 0.018 and 0.013 acts per bird/min, respectively, and standing behavior was highest. The results indicated that the frequencies of pecking inedible objects during the molt period were similar to the frequencies at 26 and 34 wk. Hens performed more acts of standing, and crouching and had lower frequency of movement during the molt. Those kept at a low density performed more movement acts. Feather pecking decreased as hens aged and increased when they molted but was not affected by strain or density. The frequency of aggression and submissive acts was significantly lower during the molt period. Behaviors were affected by strain, density, bird age, and molting; however, the patterns and number of aggressive acts did not increase to compromise the welfare status of the hens. Behaviors during the molt appeared consistent with mechanisms for conservation of body reserves. PMID- 14761080 TI - Effects of high zinc diets using zinc propionate on molt induction, organs, and postmolt egg production and quality in laying hens. AB - This study was conducted to determine the ability of an alternative salt form of 1% Zn, Zn propionate, to induce molt in 66-wk-old hens. The hens were randomly assigned to 4 treatment groups of 27 or 28 birds each: a) molted conventionally by feed withdrawal, b) 1% Zn as Zn acetate, c) 1% Zn as Zn propionate, or d) nonmolted control for 9 d. Feed intake was (P < 0.05) depressed in Zn acetate and Zn propionate hens when compared with nonmolted control hens during the 9 d. Ovary weights of hens undergoing feed withdrawal, Zn acetate, or Zn propionate were not (P > 0.05) different from each other, but all were (P < 0.05) lighter than the ovary weights of nonmolted control hens. Zinc concentrations in the kidney and liver were (P < 0.05) increased in Zn acetate and Zn propionate molted hens when compared with nonmolted hens on the control diet or hens molted by feed withdrawal. Bone ash values were (P < 0.05) increased for Zn acetate and Zn propionate molted hens or nonmolted control hens as compared with molted hens on feed withdrawal. Over the entire 3-mo postmolt period, there were no significant differences in interior egg qualities, but egg weights from hens fed Zn propionate were (P < 0.05) heavier than those from hens on feed withdrawal. The data of the current study demonstrated that feeding a Zn propionate (1% zinc) supplemented diet can induce molt. PMID- 14761081 TI - Effects of dietary zinc supplementation on broiler performance and nitrogen loss from manure. AB - An experiment was conducted to evaluate the effects of ZnSO4 or ZnO supplementation of broiler diets on growth performance and loss of uric acid N and total N from manure. A total of 240, 1-d-old broiler males were used for this experiment. Each dietary treatment was replicated 3 times with 10 birds per replicate. Chicks were fed a control diet for the first 6 d and then treatment diets for the next 12 d. There were 8 dietary treatments: the control, CuSO4-20, ZnSO4-500, ZnSO4-1,000, ZnSO4-1,500, ZnO-500, ZnO-1,000, and ZnO-1,500 containing 0, 0, 500, 1,000, 1,500 ppm supplemental Zn as ZnSO4 and 500, 1,000, and 1,500 ppm supplemental Zn as ZnO, respectively. A 300-g sample of the broiler manure from each treatment was incubated in a pan for 3 wk at room temperature. After incubation, samples were collected for the measurement of total N and uric acid N. Weight gain, feed consumption, and feed efficiency of chicks fed the diets supplemented with 1,500 ppm Zn as ZnSO4 were significantly lower than those of the other treatments, whereas the ZnO treatments had no negative effects on growth performance. After the 21-d incubation, the uric acid-N levels of manure from chicks fed the ZnO-1,000 treatment were significantly higher than those of manure from chicks fed the ZnSO4-500. The manure from chicks fed the Zn supplemented diets had significantly less total N loss compared with that from chicks fed the control. The manure from chicks fed ZnO-1,500 had significantly less total N loss than that from chicks fed the other treatment diets. This study indicated that the Zn treatments significantly reduced nitrogen loss in poultry manure, and ZnO could be a better Zn source to prevent nitrogen loss to the atmosphere without any detrimental effect on growth performance. PMID- 14761082 TI - An alternative to antibiotic-based drugs in feed for enhancing performance of broilers grown on Eimeria spp.-infected litter. AB - Three trials were conducted to evaluate the effects of lasalocid, an anticoccidial feed additive (90.7 kg/ton); bacitracin, a growth-promoter (50 g/ton); and yeast culture residue (YCR) (1 kg/ton) on the performance of broiler chicks reared to 42 d of age on recycled litter. Recycled litter consisted of pine wood shavings containing droppings from chicks infected with 3 select strains of coccidia (Eimeria tenella, Eimeria maxima, and Eimeria acervulina). Response variables (BW, intestinal tract and litter coliform counts, cecal and liver relative weights, and litter moisture content) were recorded biweekly. Mean BW of chicks fed the diet supplemented with YCR was higher than that of the controls (P < 0.05) and comparable to that of the lasalocid-treated birds in all 3 trials. Mean BW of chicks in all treatment groups decreased uniformly as the litter aged and moisture content increased. The mean intestinal coliform population from YCR-treated chicks was lower (P < 0.05) than those of the control and lasalocid populations. The coliform count was consistently lower than that in chicks on a bacitracin-supplemented diet. Coliform counts from the control and lasalocid-treated birds did not differ. The litter coliform counts increased with increased use of the litter. Cecal and liver relative weights calculated from the chicks in trial 3 showed that only the liver was significantly affected by treatments. YCR appeared to be a viable alternative to bacitracin and lasalocid medication in enhancing growth of broiler chicks reared on recycled litter. PMID- 14761083 TI - Characterization and electrotransformation of Lactobacillus crispatus isolated from chicken crop and intestine. AB - Lactic acid bacteria originating in the intestine have recently undergone intensive study for their potential probiotic properties. Here partial 16S rRNA gene sequencing of 8 Lactobacillus strains proved them to be Lactobacillus crispatus. Fatty acid analysis confirmed strains being closely related. These strains and type strain ATCC33820 were characterized for genetic engineering potential, thus determining aerobic growth, erythromycin sensitivity, and glycine tolerance. Out of 5 plasmids, a 2.9-kb plasmid (pLEB579) was successfully introduced into 4 chicken-originated wild-type L. crispatus strains. Transformation frequency was approximately 30 transformants per microgram of DNA, the first reported electrotransformation into chicken-originated L. crispatus. In spite of its low frequency, transformation enables bioengineering of these strains to improve the probiotic function in feed adsorption, chicken health, and food safety. PMID- 14761084 TI - Sequential feeding programs for broiler chickens: twenty-four- and forty-eight hour cycles. AB - Sequential feeding programs (Seq) of 2 feeds, 1 protein rich-energy poor and 1 energy rich-protein poor, during the same day or every other day were compared with a control complete diet for male broiler growth and body composition from 15 d to market weight. In experiment 1, BW gain and breast meat yield were significantly lower than those of controls for 24-h-cycle Seq with extreme protein content difference between the 2 feeds. BW gain and breast meat yield were higher than those of controls when feeds with moderate differences [feed moderately rich in protein (PM) = 26% CP; feed moderately rich in energy (EM) = 16% CP] were fed. Feeding periods that were half as long but changed twice as frequently gave less favorable results. In experiment 2, effects of allowing access to feed for different lengths of time were measured. The treatments in a 24-h cycle were a constant control diet, 50% PM-50% EM, 40% PM-60% EM, or 80% PM 20% EM decreasing to 40% PM-60% EM. Chickens fed 40% PM-60% EM had a 6% lower BW gain and a 3% higher feed/gain ratio and were fatter than those from other treatments. In experiment 3, male chickens fed a 48-h-cycle Seq of EM and PM every other day had the same BW gain, feed intake, and feed/gain ratio as controls. In experiment 4, five 48-h-cycle Seq with varying differences of energy and protein contents between EM and PM gave similar or slight reduction of performances compared with controls. A field trial with 8 flocks of broilers confirmed that feeding high and low protein feeds on alternate days resulted in performance similar to that from feeding a complete feed despite large day-to-day variations in lysine intake. Converse to shorter phases, Seq for 48-h cycles offers new opportunities for practical application and also for studies of short term regulation of protein and lipid metabolism in chickens. PMID- 14761085 TI - Effect of microbial phytase on ileal digestibility of phytate phosphorus, total phosphorus, and amino acids in a low-phosphorus diet for broilers. AB - The study aimed to assess the effect of a commercially available microbial phytase added to a corn-soybean meal diet on phytate P and total P in terminal ileal digesta as well as on true ileal amino acid digestibility. Three low-P diets containing 0, 500, or 750 U/kg of microbial phytase were fed to 21-d-old broiler chickens. Titanium dioxide was used as an indigestible marker. Ileal contents were collected from euthanized birds and analyzed, along with the diets, for total P, phytate P, and amino acids. Endogenous P determined at the terminal ileum was 446 +/- 59 mg/kg food dry matter (mean +/- SE). Endogenous ileal amino acids ranged from 219 +/- 33 mg/kg food dry matter for tryptophan to 1,255 +/- 166 mg/kg food dry matter for glutamic acid. Supplementation with microbial phytase resulted in a significantly (P < or = 0.05) greater phytate P disappearance (11% greater disappearance vs. unsupplemented control) from the terminal ileum. Similarly, true ileal total P digestibility was (P < or = 0.05) higher (10 to 12%) when microbial phytase was added. True ileal amino acid digestibility was significantly (P < or = 0.05) greater in the presence of microbial phytase for all the amino acids examined with the exception of methionine, tyrosine, histidine, and tryptophan. The mean increase in true ileal amino acid digestibility was 3.4%. The effect of phytase on true ileal phytate P, total P, and amino acid digestibility was similar for the 2 phytase inclusion levels tested. Microbial phytase improved phytate P and total P digestibility, as well as true ileal amino acid digestibility, for a corn-soybean diet. PMID- 14761086 TI - Microencapsulated short-chain fatty acids in feed modify colonization and invasion early after infection with Salmonella enteritidis in young chickens. AB - Short-chain fatty acids (SCFA) are widely used as feed additives in poultry for the control of pathogenic bacteria, such as Salmonella enteritidis. Recently, a new range of products was developed in which SCFA are encapsulated in mineral carriers, resulting in a slow release during the transport of these carriers through the intestinal tract. To test the efficacy of this type of products against early colonization after Salmonella infection in poultry, a challenge experiment with S. enteritidis was performed. Five groups of 20 chickens were given feed with no supplement or feed supplemented with acetic acid (0.24%), formic acid (0.22%), or propionic acid (0.27%) as film-coated microbeads or butyric acid (0.15%) as spray-cooled microcapsules. The 5 groups were challenged with 5 x 10(3) cfu S. enteritidis at d 5 and 6 posthatch, and samples of ceca, liver, and spleen were taken at d 8 and analyzed for the number of colony-forming units of Salmonella per gram of tissue. Feed supplementation with acetic acid, and to a lesser extent formic acid, resulted in an increase of colonization of ceca and internal organs. Birds receiving propionic acid-coated microbeads as feed supplement were colonized with Salmonella to the same extent as controls. Butyric acid-impregnated microbeads in the feed, however, resulted in a significant decrease of colonization by S. enteritidis in the ceca but not in liver and spleen. PMID- 14761087 TI - The use of low-protein, low-phosphorus, amino acid- and phytase-supplemented diets on laying hen performance and nitrogen and phosphorus excretion. AB - Two experiments were conducted to determine whether, by using a low-protein, amino acid-supplemented diet or a low-protein, amino acid-supplemented diet in conjunction with low-P, phytase-supplemented diet, the excretion of N and P could be reduced without affecting the productive performance of laying hens. Eight dietary treatments were assigned to Babcock B300 hens in each of 2 experiments that involved a positive control (16 to 16.5% CP) and a negative control (13% CP) with and without supplementation with the limiting essential amino acids. In experiment 1, supplementing the negative control with lysine, methionine, and tryptophan resulted in performance comparable to that obtained with the positive control, with the exception that egg weight was heavier for the negative control supplemented with amino acids. Supplementing the negative control with additional essential amino acids improved the performance higher than the positive control indicating that the positive control was deficient in one or more essential amino acids. In experiment 2, supplementing the negative control containing 0.2% nonphytate P (NPP) with all the limiting amino acids plus phytase resulted in performance comparable to the positive control group, which was fed 0.4% NPP without phytase. The results of a digestibility assay indicated that daily total P and N excretions of the negative control containing 0.2% NPP and supplemented with limiting amino acids and phytase were reduced by 48 and 45% of the positive control group, respectively, without compromising laying performance. PMID- 14761088 TI - The effects of supplementation of humate and probiotic on egg production and quality parameters during the late laying period in hens. AB - This study was designed to investigate whether inclusions of humate and probiotic into diets of hens during the late laying period increases egg production and improves egg quality. Hisex Brown layers (n = 300), 54 wk of age, were fed a control diet, 0.1% humate, 0.2% humate, 0.1% probiotic, or 0.2% probiotic for 75 d. Active ingredients of humate and probiotic were polymeric polyhydroxy acids (humic, fulvic, ulmic, and humatomelanic acids) and bacterial cultures (Lactobacillus, Bifidobacterium, Streptococcus, and Enterococcus spp.), respectively. Egg production and feed intake were measured daily, and egg weight was measured biweekly. Also, a sample of 12 eggs from each group was collected randomly to determine egg quality every 25 d. The data were analyzed as repeated measures with time as subplot. There were no effects of dietary treatments on feed intake and egg weight. Egg production for hens supplemented with humate and probiotic was not different but was greater than for control hens. Egg production increased linearly and mortality and feed conversion efficiency (weight of feed/weight of eggs) decreased linearly with increasing levels of supplemental humate and probiotic. There were no effects of treatments on egg quality. In conclusion, supplementation of humate and probiotic during the late laying period increased egg production, reduced mortality, and improved feed conversion efficiency but did not improve egg quality. PMID- 14761090 TI - Microbial contamination in inoculated shell eggs: II. Effects of layer strain and egg storage. AB - Three Ottawa control strains and a current commercial laying stock were reared and housed in the same environment. Eggs were collected at 5 different hen ages throughout the 2 production cycles of the flock. The eggs were inoculated with Salmonella Enteritidis (SE), Pseudomonas fluorescens (PF), a combination of the 2, or sterile buffered peptone water and stored up to 5 wk. After storage at room temperature, contamination levels were determined for the exterior surface, air cell, egg contents, and within the shell. Interior, egg contents, and shell contamination levels of SE and PF increased with storage time. There were no apparent increases in the infectivity of SE or PF in the presence of the other organism. PF was a poor survivor on the shell surface under these storage conditions. Throughout the 5-wk storage, eggs from control strain 10 maintained their microbial integrity more effectively. Eggs from control strain 5 and the current commercial stock were more easily contaminated than the other strains. These data suggest that genetic selection has altered microbiological defenses of the eggs produced. PMID- 14761089 TI - Determination of cooling rates and carbon dioxide uptake in commercially processed shell eggs using cryogenic carbon dioxide gas. AB - The ability to rapidly cool shell eggs to 7 degrees C is important in the prevention of Salmonella Enteritidis (SE) growth. In addition, quality may also be maintained longer from rapid cooling of shell eggs. A commercial cryogenic CO2 egg cooling unit was designed and installed in a commercial egg processing facility. This unit was installed on a packer head to rapidly cool eggs individually prior to packaging. The objective of this study was to determine cooling rates and CO2 gas changes that result from rapidly cooling eggs using this commercial cryogenic egg cooling system and subsequent storage for 15 wk. Results indicated that cryogenic CO2 cooling quickly cooled shell eggs in approximately 45 min, whereas traditional cooling required from 19 to 116 h. CO2 uptake into the albumen was greater in cryogenically cooled eggs (2.11 mg/g) than in traditionally cooled eggs (1.81 mg/g) immediately after processing. No differences were observed in albumen CO2 content after 2 wk of storage; at 10 wk statistically greater CO2 content remained in the cryogenically cooled eggs (1.75 mg/g) compared with the traditionally cooled eggs (1.60 mg/g). These results suggest that a large amount of CO2 enters the egg during the cryogenic cooling process but is quickly lost during storage. Beyond 10 wk of storage, the albumen CO2 content in cryogenically cooled eggs was higher than in the traditionally cooled eggs suggesting chemical changes may have occurred in the albumen. PMID- 14761091 TI - Principal component analysis of physical, color, and sensory characteristics of chicken breasts deboned at two, four, six, and twenty-four hours postmortem. AB - The effects of various postchill deboning times on functional, color, yield, and sensory attributes of broiler breast meat were determined. Broiler breast muscles were deboned at 2, 4, 6, and 24 h postmortem, and pH, color change, cooking yield, shear force values, and sensory traits of the breast meat were recorded. Data were examined by multivariate data analysis, namely principal component analysis (PCA). Averages of 13 variables (pH, delta a*, shear force, and sensory attributes of cardboardy, wet feathers, springiness, cohesiveness, hardness, moisture release, particle size, bolus size, chewiness, and metallic aftertaste afterfeel) decreased gradually as deboning time increased from 2 to 24 h, especially for shear values after 4 h of postmortem aging. Univariate correlation coefficients among 24 variables indicated several significant correlations. Warner-Bratzler shear force had high positive correlations with 5 sensory texture attributes (cohesiveness, hardness, particle size, bolus size, and chewiness). The parameters of pH, delta L*, delta a*, delta b*, and cooking yield were not obviously correlated with shear force values or any of the 18 sensory characteristics. PCA score plot showed no clear separation of the breast muscles deboned at different postmortem times, but it was still possible to differentiate them. The loading biplot suggested that 18 variables were effective in sample differentiation, including delta L*, shear force, cooking yield, 6 sensory flavor attributes (brothy, cardboardy, wet feathers, blood/serumy, salty, and sour), all sensory texture attributes except springiness, and all afterfeel-aftertaste properties. PMID- 14761092 TI - Relating induced in situ conditions of raw chicken breast meat to pinking. AB - Our objective was to simulate the pink color defect in cooked chicken breast meat with treatment combinations that would induce measurable changes in the conditions of raw meat. In addition, the feasibility of using induced raw meat conditions to develop a logistic regression model for prediction of pinking was studied. Approximately 960 breast fillets from 2 plants with 2 replications were used for inducing in situ conditions with 16 combinations of sodium chloride, sodium tripolyphosphate, sodium erythorbate, and sodium nitrite (present and not present). Muscles in all treatments were subjected to individual injections, followed by tumbling, cooking, and chilling. Raw samples were analyzed for pH, oxidation-reduction potential, and pigment evaluation. Results indicated a significant role of induced in situ conditions of raw meat in the occurrence of pinking. Presence of 1 ppm or more of sodium nitrite in raw meat produced significant pinking of cooked meat. The light muscle color group was least affected and the dark group was most affected by induced pH, oxidation-reduction potential conditions, and metmyoglobin and nitrosopigment content. The predictive ability of the logistic model was more than 90% with nitrosopigment, pH, and reducing conditions being the most important factors. Moreover, validation of the model was confirmed by close association between observed pink samples and those predicted as pink. PMID- 14761093 TI - Moisture retention by water- and air-chilled chicken broilers during processing and cutup operations. AB - The objective of this study was to evaluate effects of storage and cutting on moisture retention by air- and water-chilled broiler chickens. Sixty-four broilers were slaughtered, chilled by cold air or immersion in water, stored over night, cut into fore- and hindquarters, and then stored an additional 24 h. Air chilling conditions were 4 degrees C with air velocity of 2.2 m3/min. Water chilling conditions were 1 degree C with mechanical agitation. Moisture absorption and retention were observed as weight changes throughout the process. Air-chilled carcasses lost an average of 0.68% of their postslaughter weight in storage prior to cutting but lost no more during cutting or postcutting storage. The water-chilled carcasses absorbed 11.7% moisture in chilling but retained 6.98% through precutting storage, 6.00% through cutting and 3.90% through postcutting storage. These data offer baseline values for use in complying with new USDA processing standards. PMID- 14761094 TI - Composition, color, and texture of Thai indigenous and broiler chicken muscles. AB - Chemical compositions and physical properties of mixed-sex Thai indigenous (Gallus domesticus) and broiler (commercial breed, CP707) chicken biceps femoris and pectoralis muscles were determined. Indigenous chicken muscles contained higher protein contents but lower fat and ash contents compared to broiler muscles (P < 0.001). The amino acid profile of the indigenous chicken muscles was similar to that of the broiler muscles except they were slightly richer in glutamic acid (P < 0.05). The indigenous chicken muscles contained more saturated and less polyunsaturated fatty acids than the broiler muscles. There were no differences in the monounsaturated fatty acid contents between the breeds. The total collagen contents of indigenous pectoralis and biceps femoris muscles were 5.09 and 12.85 mg/g, respectively, which were higher than those found in broiler pectoralis (3.86 mg/g) and biceps femoris muscles (8.70 mg/g) (P < 0.001). Soluble collagen contents were lower for indigenous pectoralis and biceps femoris muscles, 22.16 vs. 31.38% and 26.06 vs. 33.87%, respectively. The CIE system values of lightness (L*), redness (a*), and yellowness (b*) of indigenous chicken muscles were higher than those of broiler muscles. The shear values of indigenous chicken muscles either raw or cooked were higher than those of broiler muscles (P < 0.05). After cooking, the shear values decreased for broiler biceps femoris and pectoralis muscles (P < 0.05), whereas no change was observed for indigenous chicken biceps femoris muscle (P > 0.05). Shear values increased for indigenous chicken pectoralis muscle (P < 0.05). PMID- 14761095 TI - Effect of vacuum on moisture absorption and retention by marinated broiler fillets. AB - Effects of vacuum on moisture retention and quality characteristics of aged chicken breast fillets were evaluated. One hundred twenty-eight broilers (2 replicates of 64 birds each) were manually slaughtered, chilled in ice water, placed in unsealed plastic bags, and then aged overnight at 4 degrees C. Both pectoralis major muscles were harvested from each carcass. Left muscles were marinated for 30 min en vacuo with 20% (vol/wt) of a 10% NaCl (wt/vol) solution containing 4% (wt/vol) commercial food-grade polyphosphate. Right fillets were marinated similarly but without vacuum. Moisture absorption, cooked yield, pH change during marination, and shear values of vacuum-marinated fillets were compared to those on fillets marinated without vacuum. Use of vacuum during marination increased moisture absorption during marination, but after cooking, yields were similar. Nor did vacuum effect pH or shear values. Under the conditions of this study, use of vacuum during marination appeared to offer no significant advantage over marination at atmospheric pressure. PMID- 14761096 TI - What's a doctor to do? Helping patients decide about prostate cancer screening. PMID- 14761097 TI - Experienced doctors, be careful what you say. PMID- 14761098 TI - Food for thought. PMID- 14761099 TI - Certification in Latvia. Turning to family medicine. PMID- 14761102 TI - Doctor-patient communication: introduction to series. PMID- 14761100 TI - Risks of untreated depression during pregnancy. AB - QUESTION: One of my patients who was taking an antidepressant for major depression is now pregnant and does not wish to take it any more. I believe she needs to continue her medication. She, however, is adamant about stopping it because she believes it would put her baby at risk. Is there evidence that not treating depression during pregnancy puts babies at risk? ANSWER: A growing body of literature investigating the effects of not treating depression on mother and developing fetus suggests that untreated depression is associated with adverse fetal outcomes and a higher risk of maternal morbidity, including suicide ideation and attempts, and postpartum depression. PMID- 14761101 TI - Dermacase. Pityriasis rosea. PMID- 14761103 TI - The medical interview: setting, nonverbal language, and social roles. PMID- 14761104 TI - Leisure activity and risk of dementia scrabble, anyone? PMID- 14761105 TI - Current status of PSA screening. Early detection of prostate cancer. AB - OBJECTIVE: To update current evidence for prostate-specific antigen (PSA) screening for prostate cancer and to give readers some practical information to discuss with patients. QUALITY OF EVIDENCE A MEDLINE: search revealed only three randomized studies, two of which are incomplete. Several controlled non randomized studies were found. MAIN MESSAGE: Two ongoing studies have not yet reported survival data, but have added to evidence for screening intervals. One Canadian randomized study has been criticized for its design and conclusions. Non randomized studies suggest that screening effectively identifies serious cancers and leads to earlier diagnosis. Mortality from prostate cancer has been falling in most western countries since 1992. This cannot be explained by PSA screening, which would probably not produce survival benefit until at least 10 years after its unofficial introduction in about 1990. CONCLUSION: Indirect evidence suggests that all men older than 45 with at least a 10-year life expectancy should be informed of the potential benefits and drawbacks of PSA screening so they can make an informed decision on whether to have the test. PMID- 14761107 TI - Case report: stroke following an airline flight. PMID- 14761106 TI - What's a man to do? Treatment options for localized prostate cancer. AB - OBJECTIVE: To describe treatments for localized prostate cancer: surgery, external radiation therapy, and brachytherapy; watchful waiting might also be appropriate. Patients trying to decide about treatment ask family physicians for advice. This article sets out a framework to aid patients (and physicians) in the decision. QUALITY OF EVIDENCE: Only two randomized studies comparing different treatments were identified. Because of the paucity of level I or II evidence, suggestions in this review are largely based on expert opinion and consensus statements. MAIN MESSAGE Risk-grouping and nomograms are useful for assessing treatments and estimating outcomes of treatment. Where treatments are equivalent, decisions can be based on perception of toxicity and convenience. Effects on patients'lives and on sexual, urinary, and bowel function vary by treatment modality. CONCLUSION: Men with low-risk prostate cancer should decide on treatment based on their perception of how treatment will affect their lives. Men with higher-risk cancers might accept adverse effects on their quality of life in return for longer survival. PMID- 14761108 TI - Assessing antenatal psychosocial health. Randomized controlled trial of two versions of the ALPHA form. AB - OBJECTIVE: To determine whether participants preferred a provider-completed or self-reported antenatal psychosocial health assessment (ALPHA) form, to evaluate the forms' effectiveness in facilitating disclosure of psychosocial issues, and to determine whether different providers gathered different information. DESIGN: Randomized controlled study. SETTING: Offices of family physicians and public health nurses (PHNs) in three health regions on Prince Edward Island. PARTICIPANTS: Physicians, PHNs, and 76 pregnant women. INTERVENTIONS: Participants completed one form and a questionnaire on theirexperience. Providers were also interviewed. MAIN OUTCOME MEASURES: Suitability and effectiveness of the forms and frequency of issues disclosed by type of form and provider. RESULTS: Most participants would recommend routine use of the ALPHA form for all pregnant women. Of the 238 psychosocial issues disclosed, significantly more were disclosed to physicians than to PHNs. CONCLUSION: Both forms were acceptable to women and providers (no clear preference emerged) and were effective at gathering information. Physicians gathered significantly more information than PHNs. PMID- 14761109 TI - Domestic violence. Incidence and prevalence in a northern emergency department. AB - OBJECTIVE: To examine the incidence and prevalence of domestic violence (DV) against women presenting to emergency departments. DESIGN: Prospective cohort study to determine health status and exposure to DV. SETTING: Hospital emergency department in urban northern Canada. PARTICIPANTS: Random sample of women older than 16 presenting to the emergency department for any reason. MAIN OUTCOME MEASURES: Demographic variables, exposure to DV. RESULTS: Of 1800 potential subjects, 577 (32%) did not fit inclusion criteria. Of the remaining 1223, 983 (80%) agreed to participate. Mean age was 41, 135 of participants (14%) were aboriginal, and 546 (56%) were married. Overall, 725 (74%) had current partners. Incidence of DV resulting in emergency department presentation on the day of assessment was 2%. Of women with partners, 66 (9%) had previously been threatened or injured by those partners. Lifetime prevalence of DV was 51%; physical DV was experienced by 40%. One-year prevalence was 26%. CONCLUSION: Incidence of DV was lower than expected; prevalence of DV was high. PMID- 14761110 TI - Combining the biologic with the psychosocial. Why family physicians conducting research should be interested. PMID- 14761111 TI - Medicine in India. Sarita's story. PMID- 14761112 TI - Looking forward, looking back. PMID- 14761113 TI - Oral treatment with SRP299 (killed Mycobacterium vaccae) inhibits experimental periodontal disease in Wistar rats. AB - OBJECTIVE: Mycobacterium vaccae injected subcutaneously was previously shown to prevent and treat ligature-induced periodontal disease (PD) in Wistar rats (Breivik & Rook 2000, 2002). Since mycobacteria are readily taken up via Peyer's patches in the intestine, we have now tested the ability of oral SRP299 (killed M. vaccae) to prevent ligature-enhanced PD in Wistar rats, and to modulate the accompanying cytokine and corticosterone responses. MATERIAL AND METHODS: A single oral dose of SRP299 (1 mg) was given 14 days before the application of ligatures. PD was assessed when the ligatures had been in place for 56 days. RESULTS: Oral SRP299 reduced bone loss (p = 0.036, X-ray; p = 0.061, histometry) and fibre loss, both on the ligatured (p = 0.0047) and control (p = 0.005) sides, and also reduced the level of TNF-alpha (p = 0.0137) and corticosterone (p = 0.048) induced by intraperitoneal endotoxin lipopolysaccharide (LPS). CONCLUSIONS: SRP299 administered by the oral route diminishes ligature-induced bone and fibre loss in this model. This effect may be attributable to the known ability of SRP299 to evoke regulatory T cells, although the mechanism could not be investigated in this study. Treated rats also had less excitable hypothalamo pituitary-adrenal (HPA) axes. HPA axis overactivity is a known risk factor in human PD. Trials of SRP299 in human PD are now justified. PMID- 14761114 TI - Expression of RNAs encoding for alpha and beta integrin subunits in periodontitis and in cyclosporin A gingival overgrowth. AB - BACKGROUND: Variation of integrin expression in healthy and diseased gingiva revealed a potential biological role for these cell matrix receptors during gingival remodeling. AIM: Here we determined the level of RNA and tissue localization of different integrin subunits in periodontitis and cyclosporin A induced gingival overgrowth. METHODS: The level of expression was determined by Reverse Transcriptase Polymerase Chain Reaction in 12 periodontitis-affected patients, four patients exhibiting severe cyclosporin A-induced gingival overgrowth and seven healthy patients as controls. RESULTS: The RNA encoding for beta1, alpha2 and alpha5 integrin subunits were reduced in periodontitis gingiva. The reduction observed was stronger in cyclosporin A-treated patients as compared to the healthy controls, while RNA encoding for alpha1 subunit was increased. The RNA encoding for alpha6 integrin was only reduced in cyclosporin A-treated gingiva. Immunohistochemistry showed that i) integrin alpha2 expression is restricted to the gingival epithelium of cyclosporin A-treated patients, ii) the reduction of alpha6 integrin expression in cyclosporin A-treated gingiva is due to loss of expression at focal contacts and iii) beta1 integrin is evenly distributed in the three populations with an intensity decrease in periodontitis and cyclosporin A-treated gingiva. CONCLUSION: Taken together these results showed a role for the integrin receptors in periodontal diseases and cyclosporin A-induced gingival overgrowth. PMID- 14761115 TI - Longitudinal evaluation of GCF IFN-gamma levels and periodontal status in HIV+ patients. AB - BACKGROUND/AIM: Loss of periodontal support and related tooth loss is a common finding among HIV+ patients. The etiology of this destruction may be an increase in the levels of pro-inflammatory cytokines and subsequent increase in periodontal disease activity. The purpose of this study was to investigate the associations between gingival crevicular fluid interferon gamma (GCF IFN-gamma) and clinical measures of periodontal disease in HIV+ individuals. We monitored GCF IFN-gamma and periodontal status of selected sites in 33 HIV+ subjects over a 6-month period. METHOD: Clinical measurements including gingival index, plaque index, bleeding on probing, probing depth, attachment loss (AL), and GCF samples were taken from four lower incisors and the upper right posterior sextant of each patient at baseline and 6-month visits by means of sterile paper strips. GCF levels of IFN-gamma were determined by sandwich ELISA assays. A progressing site was defined as a site that had 2 mm or more AL during the 6-month study period. RESULTS: Twenty-five of the 264 examination sites showed 2 mm or more clinical AL during the 6-month study period. Significantly higher GCF levels of IFN-gamma were found at progressing sites than in nonprogressing sites (p < 0.001). GCF levels of IFN-gamma were highly correlated with clinical measurements taken at baseline and 6-month visits (0.001 0.05). The uronic acid content of GCF and gingiva showed significant overlaps between MP and SP sites and uronic acid levels did not present any constant correlation with the clinical parameters (p > 0.05). In a similar manner, uronic acid content of GCF and gingival tissue was not correlated (p > 0.05). CONCLUSION: The lack of a significant correlation between the uronic acid content of gingival tissue and GCF may suggest that the passage of PG metabolites from gingiva to GCF is likely to be under the influence of multifactorial interactions rather than being linear. As a general measure of PG metabolism, uronic acid levels do not seem to be related with the severity of periodontal destruction and tend to act as different measures when compared to traditional clinical parameters. PMID- 14761119 TI - Periodontal conditions in patients with juvenile idiopathic arthritis. AB - OBJECTIVE: Our aim was to compare the periodontal conditions in a group of juvenile idiopathic arthritis (JIA) patients with those in a control group of healthy subjects (CTR). MATERIAL AND METHODS: Thirty-two patients with JIA and 24 controls were selected. The measurements used to diagnose periodontal disease included plaque and bleeding scores, probing depths (PDs) and clinical attachment loss (CAL). Laboratory indicators of JIA activity included the erythrocyte sedimentation rate (ESR) and capsule-reactive protein (CRP). The Mann-Whitney test was used to evaluate the data (alpha = 0.05). RESULTS: The mean ages were 15.9 (+/- 2.7) years and 14.7 (+/- 2.3) years for groups JIA and CTR, respectively. The median ESR was 42 mm/h 13 mm/h in the CTR group (p = 0.032) and the median CRP was 1.9 and 0.4 mg/l, respectively (p = 0.001). The prevalence of patients with a proximal attachment loss of 2mm or more in the JIA group was 25% and in controls it was 4.2%. The mean percentages of visible plaque and marginal bleeding were similar in the JIA (54 +/- 22 and 30 +/- 16, respectively) and CTR groups (44 +/- 18 and 29 +/- 11, respectively). The mean percentages of sites with PD > or = 4 mm were significantly higher in the JIA group (3 +/- 4.7) than in the CTR group (0.4 +/- 1.7) (p = 0.012). The mean percentages of sites with proximal CAL > or = 2 mm were 0.7 (+/- 1.4) in the JIA group and 0.001 (+/- 0.2) in the CTR group (p = 0.022). CONCLUSION: Adolescents with JIA present more periodontal attachment loss than healthy controls, in spite of similar plaque and marginal bleeding levels. PMID- 14761121 TI - Perceived risk of deteriorating periodontal conditions. AB - BACKGROUND: Interpretation of risk for periodontitis is critical for treatment planning. How periodontists assess risk for periodontitis is unclear. PURPOSE: To study (1) what factors periodontists use when assessing the risks for worsening periodontal conditions anticipating that no treatment would be provided, and (2) if risk assessment is consistent and independent of specialty background training. MATERIAL AND METHODS: Medical history, clinical dental data, full-mouth intra-oral radiographs, and slide pictures were obtained from each of 51 subjects, and the information was provided to 23 examiners. RESULTS: The mean age of the study subjects was 51.5 years (SD +/- 17.7, range 23-81), with 28 women included. In 10 of the subjects, only gingivitis was identified, while 22 subjects had advanced chronic periodontitis. Risk scores assigned for 2 and 4 years differed significantly between European- and US-trained periodontists (p < 0.001) and between graduate students in training and periodontists from either the US or Europe (p < 0.01) (Wilcoxon n-pair test), with European periodontists scoring the lowest risks. Risk scores were correlated between groups (p < 0.01 with rho range 0.82-0.89) (Spearman's rank correlation). The best-fit model (r2 = 0.86) to assess perceived risk for worsening periodontal conditions based on data from all examiners combined included the following variables: (1) overall horizontal alveolar bone loss (p < 0.000), (2) age-adjusted proportional radiographic bone height score for the worst site (p < 0.000), and (3) proportion of pocket probing depths > or = 6.0 mm. CONCLUSIONS: Differences exist on the scale of risk values based on specialty training. Consistency in scoring patterns exists. The examiners based their assigned risk scores almost exclusively on measures of existing disease severity, including radiographic bone loss and numbers of periodontal pockets > or = 6.0 mm, and excluding most known risk factors such as smoking, diabetes, and poor oral hygiene. PMID- 14761120 TI - Clinical evaluation of the Er:YAG laser in combination with an enamel matrix protein derivative for the treatment of intrabony periodontal defects: a pilot study. AB - OBJECTIVES: The aim of the present study was to compare the combination therapy of deep intrabony periodontal defects using an Er:YAG laser (ERL) and enamel matrix protein derivative (EMD) to scaling and root planing+ ethylenediaminetetraacetic acid (EDTA)+EMD. MATERIAL AND METHODS: Twenty-two patients with chronic periodontitis, each of whom displayed 1 intrabony defect, were randomly treated with access flap surgery and defect debridement with an Er:YAG (160 mJ/pulse, 10 Hz) plus EMD (test) or with access flap surgery followed by scaling and root planing (SRP) with hand instruments plus EDTA and EMD (control). The following clinical parameters were recorded at baseline and at 6 months: plaque index, gingival index, bleeding on probing (BOP), probing depth (PD), gingival recession, and clinical attachment level (CAL). No differences in any of the investigated parameters were observed at baseline between the two groups. RESULTS: Healing was uneventful in all patients. At 6 months after therapy, the sites treated with ERL and EMD showed a reduction in mean PD from 8.6 +/- 1.2 mm to 4.6 +/- 0.8 mm and a change in mean CAL from 10.7 +/- 1.3 mm to 7.5 +/- 1.4 mm (p < 0.001). In the group treated with SRP+EDTA+EMD, the mean PD was reduced from 8.1 +/- 0.8 mm to 4.0 +/- 0.5 mm and the mean CAL changed from 10.4 +/- 1.1 mm to 7.1 +/- 1.2 mm (p < 0.001). No statistically significant differences in any of the investigated parameters were observed between the test and control group. CONCLUSION: Within the limits of the present study, it may be concluded that both therapies led to short-term improvements of the investigated clinical parameters, and the combination of ERL and EMD does not seem to improve the clinical outcome of the therapy additionally compared to SRP+EDTA+EMD. PMID- 14761122 TI - Bacterial colonization during de novo plaque formation. AB - OBJECTIVE: To determine microbial changes that occur during plaque formation in a dentition free of gingival inflammation. MATERIAL AND METHODS: Ten subjects were recruited. The study included one preparatory period (2 weeks) and a plaque accumulation period (4 days). The volunteers exercised proper tooth cleaning methods, were scaled and received repeated professional mechanical tooth cleaning during the preparatory period. During the plaque accumulation period, the participants abstained from plaque control measures. Plaque was scored on the approximal surfaces of maxillary and mandibular premolars on Days 0, 1, 2 and 4 using a scale from 0 to 5 and according to the criteria of the Quigley and Hein Plaque Index (QHI). Supragingival plaque samples were obtained from the same intervals and surfaces and evaluated using a checkerboard DNA-DNA hybridization technique. RESULTS: The mean QHI increased from 0 to 1.6 (Day 4). The total number of organisms on Day 0 averaged 140 x 10(5) and increased to about 210 x 10(5) after 4 days without oral hygiene. The most dominant species on Day 0 were members of the genus Actinomyces. These organisms comprised almost 50% of the microbiota evaluated. None of the Actinomyces species increased significantly during the 4 days. Some Streptococcus species increased significantly over time as well as species of the genera Capnocytophaga, Campylobacter, Fusobacteria and Actinomyces actinomycetemcomitans. CONCLUSION: In the present investigation, the preparatory phase established a situation with minimal gingival inflammation and close to zero amounts of dental plaque. The Day 0 plaque samples exhibited high proportions of Actinomyces species. During the 4 days of no oral hygiene, there was a small increase in total numbers of organisms as well as a modest increase in the proportion of "disease-associated" taxa such as species of the "orange complex" species. PMID- 14761123 TI - Effect of smoking on gingival crevicular fluid cytokine profile during experimental gingivitis. AB - BACKGROUND: Cigarette smoking is a significant risk factor in the pathogenesis of periodontal disease, able to influence both the subgingival microbiota and host responses. AIM: The aim of the present study was to determine the influence of smoking on the amount of IL-1beta, IL-4 and IL-8 in gingival crevicular fluid (GCF) during experimental gingivitis in humans. MATERIAL AND METHODS: Twenty-two healthy subjects, 10 smokers and 12 non-smokers, participated in the study. After professional cleaning, they performed optimal hygiene to reach perfect clinical gingival health. Oral hygiene measures were ceased for a period of 10 days. Clinical indices, including plaque index (PI), gingival index (GI), probing pocket depth (PPD) and bleeding on probing (BOP), were assessed 2 days before (day -2), at the beginning (day 0) and at the end of the experimental gingivitis period (day 10). At the same time, GCF was collected from 12 sites in each patient, by means of durapore filter membranes. Total amounts of IL-1beta, IL-4 and IL-8 were determined by enzyme-linked immunoadsorbent assay. RESULTS: Clinical data revealed that both smokers and non-smokers showed an increase in PI, GI and BOP scores during the experiment. Although no differences were noted with regard to PI at day 10, the GI and BOP were significantly less pronounced in smokers than non-smokers (p < 0.005). Non-smokers showed higher total amounts of IL-4 but lower amounts of IL-8 than smokers, throughout the experiment. Total amounts of IL-1beta and IL-8 increased significantly during plaque accumulation in both groups. IL-4 remained stable for the smoker group and decreased for the non-smoker group. CONCLUSIONS: The present results indicate that smoking interferes with cytokine production. When performing studies regarding the pathogenesis of periodontitis, the smoking status of the participants needs to be taken into consideration. PMID- 14761124 TI - Relationship between periodontal pocket sulfide levels and subgingival species. AB - BACKGROUND: Many species implicated in the pathogenesis of periodontal disease produce volatile sulfur compounds (VSC). This investigation examined the relationship between levels of sulfide and subgingival bacterial species in the same periodontal pockets. MATERIAL AND METHODS: Twenty chronic periodontitis subjects were measured clinically at six sites per tooth for plaque, gingivitis, bleeding on probing, suppuration, pocket depth and attachment level. Subgingival plaque samples, taken from the mesial aspect of each tooth, were individually analyzed for their content of 40 bacterial species using checkerboard DNA-DNA hybridization. Sulfide levels were measured at the same sites using a Diamond Probe/Perio 2000 system. Clinical and microbiological data were averaged for sulfide-positive and -negative sites separately in each subject and then averaged across subjects. Significance differences in clinical and microbial parameters between sulfide-positive and -negative sites were sought using the Wilcoxon signed ranks test. RESULTS: Mean total DNA probe counts (x10(5), +/-SEM) at sulfide-negative and -positive sites were 44.0 +/- 9.9 and 65.0 +/- 13.3, respectively (p < 0.01). Seventeen species were found at significantly higher levels in sulfide-positive than -negative sites. These included abundant producers of VSC such as members of the genera Fusobacterium, Campylobacter, Prevotella, Treponema and Eubacterium, and Bacteriodes forsythus, Selenomonas noxia and Propionibacterium acnes. Prevotella intermedia, Bacteriodes forsythus, Prevotella nigrescens, Fusobacterium nucleatum ss vincentii and Treponema denticola exhibited the greatest difference in mean counts between sulfide negative and -positive sites. Orange and red complex species were at higher counts at shallow (< 4 mm) sulfide-positive than shallow sulfide-negative sites. Although not statistically significant, mean clinical parameters were somewhat higher at sulfide-positive than sulfide-negative sites. CONCLUSIONS: Intra-pocket sulfide levels reflect the levels of sulfide-producing species and may provide useful diagnostic information. PMID- 14761125 TI - Acellular dermal matrix allograft used to gain attached gingiva in a case of epidermolysis bullosa. AB - BACKGROUND: Epidermolysis bullosa (EB) is an acquired disease or inherited as either autosomal dominant or recessive with an incidence of 1/50,000. The prominent clinical characteristic of the disease is the development of bullae or vesicles in mucosa or skin in response to minor trauma. AIM: A female patient with a dystrophic type of EB had been put in a maintenance regimen after completion of the initial phase of periodontal therapy and followed for 7 years. The purpose of this report is to document acellular dermal matrix allograft application to increase the width of the attached gingiva in this patient experiencing difficulty in chewing and performing plaque control due to the dramatic loss of attached gingiva after 7 years of supportive periodontal therapy. METHODS: Under local anaesthesia and antibiotic coverage, the acellular dermal matrix allograft was applied in the anterior region of the upper jaw in order to increase the width of attached gingiva, thereby improving patient comfort. RESULTS: The healing was uneventful and a significant gain in attached gingiva dimensions was observed 9 months after the periodontal surgery. The procedure avoided a second surgical site, provided satisfactory results from an aesthetic point of view, and improved patient comfort. CONCLUSION: Acellular dermal matrix allograft may be regarded as an alternative in the treatment of EB cases to increase the width of attached gingiva and facilitate maintenance of the dentition. PMID- 14761126 TI - Treatment of bullous pemphigoid: what's new? PMID- 14761127 TI - Incidence of psychiatric disorders in dermatological patients. PMID- 14761128 TI - The efficacy and safety of terbinafine in children. AB - Terbinafine is an allylamine antifungal agent that has been effective and safe in the treatment of superficial and some deep mycotic infections in adults. An increasing amount of data is available where terbinafine has been used in the paediatric population to treat superficial fungal infections, in particular tinea capitis. The data suggest that terbinafine is effective and safe using treatment regimens that involve short duration therapy, leading to an increased compliance and providing a cost-effective means of treating paediatric superficial fungal infections such as tinea capitis. Terbinafine has been approved for the treatment of tinea capitis in many countries worldwide, and provides good efficacy rates for Trichophyton tinea capitis using shorter regimens than the gold standard griseofulvin. The adverse events profile for children is similar to that in adults with few adverse effects associated with its use. The evidence favours the use of terbinafine in the treatment of superficial infections in children. PMID- 14761129 TI - Influence of intravenous immunoglobulin therapy on autoantibody titres to BP Ag1 and BP Ag2 in patients with bullous pemphigoid. AB - BACKGROUND: Bullous pemphigoid (BP) is a subepidermal autoimmune blistering disease, which is characterized by blisters on the skin. Autoantibodies to components of the basement membrane zone are usually observed in the sera of patients with BP. Autoantibodies to the bullous pemphigoid antigens (BP Ag1, 230 kDa desmoplakin protein, and BP Ag2, 180-kDa hemidesmosomal protein) are present in the sera of BP patients. OBJECTIVE: The objective of this study was to report the influence of intravenous immunoglobulin (IVIg) therapy on autoantibody titres to BP Ag1 and BP Ag2. METHODS: In this prospective study, we measured autoantibody titres to both BP Ag1 and BP Ag2, in 10 patients with severe BP, over a period of 18 consecutive months on each patient, using an immunoblot assay. RESULTS: Prior to the initiation of IVIg therapy, the sera of nine patients demonstrated the presence of high autoantibody titres to both BP Ag1 and BP Ag2. One patient had autoantibodies to BP Ag1 only. A statistically significant decline in the autoantibody titres to both BP Ag1 and BP Ag2 was observed after 3 months of receiving the first cycle of IVIg therapy. This gradual decline in autoantibody titres continued until patients were observed to have non-detectable titres to BP Ag1 after 11 months and to BP Ag2 after 10 months of receiving IVIg therapy. Once patients achieved non-detectable titres, these patients were considered to be in a serological remission. This serological remission was sustained for an additional 7 months of observation. CONCLUSION: Autoantibody titres to BP Ag1 and BP Ag2 can be used to monitor the serological response to treatment in patients with BP. Patients with severe BP who are treated with IVIg therapy, as described in our protocol, achieve a long-term serological remission. PMID- 14761130 TI - In vitro effects of ultraviolet A on histamine release from human basophils. AB - BACKGROUND: As long-wave ultraviolet (UV) radiation penetrates the dermis, connective tissue cellular components and circulating blood cells can be possible targets for solar UVA. Basophils, involved in the effector phase of the inflammatory response, play a part in skin diseases such as chronic urticaria, psoriasis, atopic dermatitis, fixed drug eruption, allergic contact dermatitis, urticaria pigmentosa, systemic sclerosis and bullous pemphigoid. OBJECTIVE: The evaluation of the in vitro effect of UVA on histamine release from human basophils. METHODS: Basophils from healthy human volunteers were irradiated, respectively, with UVA at doses of 2.5, 5, 7.5, 10, 20 and 50 J/cm2 and then incubated with an anti-IgE serum. A fluorimetric technique was employed to determine histamine release from samples: (i) incubated with 2% HClO4 (complete lysis of basophils); (ii) irradiated with increasing doses of UVA; and (iii) unirradiated (controls). RESULTS: Histamine release was: 100% for HClO4 incubated basophils, 30% for unirradiated and anti-IgE incubated cells (controls) and 27%, 24%, 34%, 41%, 60% and 70% for basophils irradiated with UVA doses, respectively, of 2.5, 5, 7.5, 10, 20 and 50 J/cm2 and incubated with anti-IgE. Histamine releasability from irradiated samples was statistically significant (P < 0.05), in comparison with controls, at UVA doses equal to 5, 10, 20 and 50 J/cm2. CONCLUSIONS: UVA exerts, at least in vitro, a biphasic dose-dependent action on histamine release from human basophils incubated with an anti-IgE serum: at the lowest irradiation doses (< 5 J/cm2) it exerts an inhibitory effect and at the highest doses (> or = 10 J/cm2) histamine release increases significantly. PMID- 14761132 TI - Intertriginous lesions in pityriasis versicolor. AB - BACKGROUND: The well-recognized sites of involvement in pityriasis versicolor are the upper part of the trunk, the neck and the upper arms. There is little documentation regarding pityriasis versicolor lesions in flexural areas. OBJECTIVES: To study the occurrence and morphology of lesions of pityriasis versicolor in flexural areas. METHODS: One hundred and ten cases of pityriasis versicolor were studied. Sites of lesions and other clinical features were determined. RESULTS: Flexural lesions in pityriasis versicolor were not uncommon. They were not correlated to age, sex, duration or relapses of the infection. CONCLUSION: Thorough inspection and examination of the skin by Wood's light before establishing treatment is prudent in cases of pityriasis versicolor located in typical sites. PMID- 14761131 TI - Assessment of information and education about topical corticosteroids in dermatology outpatient departments: experience from Turkey. AB - BACKGROUND: Topical corticosteroids are commonly used in dermatological practice. The aim of this study was to assess the baseline level of knowledge about topical steroids in a group of dermatology patients and to evaluate the impact of educational materials on patients' level of information. METHODS: A 10-item questionnaire was developed, and 126 patients who were prescribed topical steroids for several dermatologic diseases were asked mostly about side-effects of the drugs. Three different educational methods were used in parallel to the questionnaire: verbal (n = 42), written (n = 42) or verbal and written (n = 42). The same questions were asked again 1 or 2 weeks later. RESULTS: Only 7.1% of the study group mentioned that they were informed by doctors about the side-effects of topical steroids before they were administered. The baseline information scores about topical corticosteroids were higher in high school and college graduates and in those who noted side-effects or who were informed about side effects before administration. A statistically significant difference was detected between pre- and posteducation scores of patients in the whole group and within the three groups of different educational methods (P < 0.001). Posteducation scores of the combined verbal- and written-educated group were higher than the written-educated group or the verbal-educated group. CONCLUSION: The present study indicated that patient knowledge is enhanced by any kind of educational method, and particularly written information supported by verbal instructions given in dermatology outpatient departments. Moreover, we conclude that the 10-item questionnaire can be used in Turkey to assess patients' knowledge about topical corticosteroids. PMID- 14761133 TI - Antioxidant activity, lipid peroxidation and skin diseases. What's new. AB - Due to its interface function between the body and the environment, the skin is chronically exposed to both endogenous and environmental pro-oxidant agents, leading to the harmful generation of reactive oxygen species (ROS). There is compelling evidence that oxidative stress is involved in the damage of cellular constituents, such as DNA, cell membrane lipids or proteins. To protect the skin against the over-load of oxidant species, it contains a well-organised system of both chemical and enzymatic antioxidant which are able to work in a synergistic manner. Skin antioxidant network protects cells against oxidative injury and prevent the production of oxidation products, such as 4-hydroxy-2-nonenal or malonaldehyde, which are able to induce protein damage, apoptosis or release of pro-inflammatory mediators, such as cytokines. When oxidative stress overwhelms the skin antioxidant capacity the subsequent modification of cellular redox apparatus leads to an alteration of cell homeostasis and a generation of degenerative processes. Topical application or oral administration of antioxidants has been recently suggested as preventive therapy for skin photoaging and UV-induced cancer. The recognition that ROS can act as second messengers in the induction of several biological responses, such as the activation of NF-kB or AP-1, the generation of cytokines, the modulation of signalling pathways, etc., has led many researchers to focus on the possible effects of antioxidants in many pathological processes. The recent demonstration that the peroxisome proliferators-activated receptors, whose natural ligands are polyunsaturated fatty acids and theirs oxidation products, have a central role in the induction of some skin diseases, such as psoriasis or acne, has indicated new links between free radicals and skin inflammation. Based on these findings, the review summarises the possible correlations between antioxidant imbalance, lipid oxidative breakage and skin diseases, from both a pathological and therapeutic points of view. PMID- 14761135 TI - Fabry disease: enzyme replacement therapy. AB - Fabry disease is a multisystem disorder associated with wide variability in clinical expression. Fabry disease is an X-linked lysosomal storage disorder caused by a deficiency of alpha-galactosidase A. The enzyme defect leads to the systemic accumulation of glycosphingolipids with alpha-galactosyl moieties consisting predominantly of globotriaosylceramide, galabiosylceramide and two additional glycosphingolipids. Four hemizygotes patients with a family history of Fabry disease and deficiency of the enzyme alpha-galactosidase A were selected. Each patient received purified alpha-galactosidase by intravenous infusion (0.2 mg/kg). The infusion was administered every 2 weeks, for 40 min, for a total of 12 months. Outcome measures include neurological manifestations (acroparaesthesia, hypohidrosis, vasomotion), kidney function, cardiac manifestations, angiokeratomas, and corneal dystrophy. alpha-Galactosidase A prepared from human fibroblast is safe and well tolerated. After 12 months of therapy the mean creatinine clearance increased, there was significant improvement in the acroparaesthesias and in the hypohidrosis. Physical stigmata, such as angiokeratomas in the skin, and characteristic benign corneal abnormalities remained stable. Enzyme replacement therapy would therefore represent a significant advance in treatment of patients with Fabry disease. Enzyme replacement therapy is safe and likely to improve the prognosis of Fabry disease. PMID- 14761134 TI - Stressful life events, anxiety, depression and coping mechanisms in patients with Behcet's disease. AB - BACKGROUND: Behcet's disease is a systemic immunoinflammatory disease of young adults characterized by systemic vasculitis of arteries and veins. Although many studies have been published since its discovery in 1937, the etiopathogenesis of this unique disorder is still unclear. OBJECTIVE: To assess the relationship between stress factors, psychological and somatic symptoms, and coping mechanisms in patients with Behcet's disease. METHOD: Thirty-four patients with Behcet's disease and 43 control subjects were compared by using sociodemographic data collection forms, a psychosocial and environmental problems list, the Beck Anxiety Inventory (BAI), Hamilton Depression Rating Scale (HAM-D) and Toronto Alexithymia Scale (TAS). RESULTS: Twenty-four patients (70.6%) defined stress factors in the first stage of the disease. Twenty-seven (79.4%) out of 34 patients stated that the recurrence period of the disease was related to the stress factors. Fear was expressed by 10 (29.4%) patients, sadness by 11 (32.3%), and fear plus sadness by 13 (38.2%) when they first learnt the diagnosis. While coping with these emotions 14 (41.2%) revealed active-reliance strategy. A statistically significant difference was present between the Behcet's patients and control subjects regarding TAS (P < 0.05), HAM-D (P < 0.001) and BAI (P < 0.001) scores. CONCLUSION: It seems that stressful life events have important implications in both relapsing and remission periods of Behcet's disease via secondary problems. PMID- 14761136 TI - A new surgical approach for the treatment of severe epithelial skin sun-induced damage. AB - BACKGROUND: Cutaneous photoageing is a complex biological process affecting all layers of the skin. Skin damage resulting from intrinsic ageing and extrinsic photoageing may trigger skin cancer. In patients with advanced photoageing and/or diffuse actinic damage, local therapy is often inadequate and the possibility of combined therapy needs to be assessed. SUBJECTS: Here we report three cases of patients over 75 years of age with advanced diffuse epithelial skin damage of photoexposed areas consisting of several superficial actinic keratoses, ipermelanotic lesions and multiple skin cancers. METHODS: Neoplastic lesions and damaged skin were removed by superficial erbium laser ablation and the epidermis reconstructed with autologous epidermal sheets expanded in vitro from healthy cells obtained from unexposed areas of the body. RESULTS: Our initial studies show that this procedure is very effective in the short term for treating and preventing the UV-induced skin cancer and precancerous lesions, and also suggest good long-term control of the disease with very interesting aesthetic results. PMID- 14761137 TI - Vulval lymphangiomata mimicking genital warts. AB - Genital warts can mimic a variety of conditions. We report a middle-aged pregnant woman with lymphangiectatic lesions on the vulva resembling warts. PMID- 14761138 TI - Granuloma annulare in a site of healed herpes zoster: Wolf's isotopic response. AB - A case of granuloma annulare that developed in a site of healed herpes zoster is reported. Polymerase chain reaction failed to detect VZV DNA in a biopsy specimen. The occurrence of different types of cutaneous reactions in a body area previously affected by herpes virus infection is known as Wolf's isotopic response. Pathogenesis may be due to a local neuroimmune dysregulation set off by herpesvirus-induced lesions of dermal sensory nerve fibres. PMID- 14761139 TI - Dissecting cellulitis of the scalp with associated spondylarthropathy: case report and review. AB - Arthritis is a well-recognized but uncommon accompaniment to several chronic cutaneous inflammatory conditions in which severe acne is one component. We report the case of a man with dissecting cellulitis of the scalp who developed severe peripheral and axial arthritis. PMID- 14761140 TI - Red fingers syndrome and recurrent panniculitis in a patient with chronic hepatitis C. AB - Cases of red fingers syndrome and nodular vasculitis in patients with hepatitis C have rarely been described. Both red fingers syndrome and nodular vasculitis are characterized by microscopic vasculitis, and it has been hypothesized that hepatitis C plays a role in the pathogenesis of cutaneous disorders such as vasculitis, especially in cryoglobulinaemic vasculitis. We describe the case of a 75-year-old woman diagnosed with chronic hepatitis C who presented with nodular lesions on her right thigh and red fingers syndrome. A skin biopsy taken from a nodule showed infiltration of the vessel walls by mononuclear cells and septal and lobular panniculitis. The diagnosis of nodular vasculitis was established. In our opinion, red fingers syndrome and nodular vasculitis might be related to a vascular reaction of immune-mediated mechanisms induced by hepatitis C. PMID- 14761141 TI - Rheumatoid nodulosis: report of two cases. AB - We present two patients with subcutaneous nodular lesions of several years' duration. These lesions were well defined and painless and were located next to bony surfaces or in the pulps of fingers and elbows. No other symptoms were observed except for frequent attacks of palindromic rheumatism with no evidence of arthritis or systemic manifestations suggestive of rheumatoid arthritis. Other studies performed were normal with negative results for rheumatoid factor and antinuclear antibodies. Radiographs of affected sites showed no signs of arthritis while the histopathologic study revealed some findings compatible with rheumatoid nodule. One of the patients also presented desquamate papular lesions at the level of elbows while histologically a process of transepidermal elimination of fibrinoid could be observed. Our patients fulfil all the diagnostic criteria for rheumatoid nodulosis as they present multiple rheumatoid nodules confirmed on biopsy, symptoms of palindromic rheumatism, no evidence of arthritis or radiological alterations and negative rheumatoid factor. PMID- 14761142 TI - Erythema multiforme ID reaction in atypical dermatophytosis: a case report. AB - BACKGROUND: Erythema multiforme (EM) is an uncommon mucocutaneous disorder, characterized by widespread erythematous-oedematous targetoid lesions, considered as a reaction pattern to many different stimuli. OBSERVATION: Presentation is made of a classical EM eruption concomitant to an inflammatory, atypical dermatophytosis of the ala nasi, observed in a 37-year-old male at the Dermatology Department of Cagliari, Italy. METHODS: Laboratory investigations revealed normal complete blood count and biochemistry profile. A direct microscopic examination with 20% potassium hydroxide of the nasal skin scales and of the vibrissae indicated the presence of dermatophyte hyphae and spores invading the rhinothrix. Culture confirmed a Trycophyton mentagrophytes infection. Specific determination of antiviral immunity in the blood (TORCH) was negative for recent infections, while the cell-mediated response, investigated with a delayed skin multitest, showed a marked positivity for the tricophyton antigen. Systemic therapy with terbinafine (250 mg/day) led to complete clinical and mycological recovery of the atypical ringworm in 30 days. Multiforme lesions rapidly improved and completely disappeared in 7 days. CONCLUSIONS: ID reaction to a distant focus of dermatophyte infection very rarely presents as EM. The presented case fits all the essential criteria required for the diagnosis of an ID reaction. The atypical clinical presentation of the ringworm infection, with marked involvement of the intranasal hairs, seems to have induced an inflammatory allergic response, similar to that evoked by sycosis. PMID- 14761143 TI - Langerhans' cell histiocytosis and haemophagocytic lymphohistiocytosis in an elderly patient. AB - We present a case of a 78-year-old man suffering from a chronic psoriasiform eruption, with rapid deterioration over the previous 8 weeks. Langerhans' cell histiocytosis with skin and bone involvement was diagnosed, and there was evidence of liver and lung dysfunction. The patient was treated with prednisolone and etoposide, and initially experienced a partial improvement. Three weeks later, haemophagocytic lymphohistiocytosis and subsequently a large pulmonary abscess with sepsis attributed to opportunistic gram-negative enterobacteriaceae Serratia marcescens developed, and the patient died. The present case of Langerhans' cell histiocytosis is of particular interest because of the previously unreported development of haemophagocytic lymphohistiocytosis in the elderly population. PMID- 14761144 TI - Lupus vulgaris: unusual presentations over the face. AB - Lupus vulgaris (LV) is the most common morphological variant of cutaneous tuberculosis. However, the occurrence of bizarre clinical presentations over atypical sites often leads to misdiagnosis and inappropriate treatment causing significant morbidity. This report seeks to highlight two unusual cases of lupus vulgaris occurring on the face of immunocompetent women and remarkably mimicking periorbital cellulitis and basal cell carcinoma, respectively. The diagnosis was confirmed by histopathology, an enzyme-linked immunosorbent assay (ELISA) test for Mycobacterium tuberculosis and polymerase chain reaction (PCR). With four drug antitubercular therapy, both patients had a dramatic response. PMID- 14761145 TI - Job's syndrome--a case report. AB - Job's syndrome is a rare immune disorder characterized by atopic dermatitis-like skin lesions, elevated serum IgE-levels, repeated occurrence of skin and respiratory tract infections, and skeletal abnormalities. We report on a 12-year old boy with Job's syndrome from Gujarat State, India. He disclosed the characteristic face, eczematous skin reactions and skin and lung infections. Long term chemoprophylaxis was realized with oral penicillins that dramatically improved the course of his disease. Other treatment options are discussed but not all meet the needs of developing countries. PMID- 14761146 TI - Undifferentiated connective tissue disease and its cutaneous manifestations. AB - Undifferentiated connective tissue disease (UCTD, also named UCT syndrome, latent lupus or incomplete lupus) is regarded as an autoimmune disorder in which signs and symptoms are widely variable and evocative for connectivitis but not sufficiently evolved to fulfil any of the accepted classification criteria for the defined connective tissue diseases. In this paper we describe the case of a 47-year-old woman affected by UCTD according to the preliminary classification criteria supplied by Mosca et al. in 1999. PMID- 14761147 TI - Familial urticaria pigmentosa associated with thrombocytosis as the initial symptom of systemic mastocytosis and Down's syndrome. AB - Most cases of urticaria pigmentosa are confined to the skin, but visceral involvement and/or haematological abnormalities have been observed. It is still a matter of debate whether all forms of mastocytosis are true neoplasias or reactive hyperplasias. Familial inheritance of urticaria pigmentosa is rare. We report on a fraternal set with urticaria pigmentosa as part of a systemic mastocytosis. The first patient additionally revealed persistent thrombocytosis and splenomegaly. His brother developed urticaria pigmentosa, intermittent diarrhoea, hepatomegaly and asthma bronchiale associated with trisomy 21 (Down's syndrome). The association of mastocytosis with thrombocytosis has seldom been described. In our patient it preceded the development of systemic mastocytosis. The association with Down's syndrome has not been reported until now. PMID- 14761148 TI - Disseminated molluscum contagiosum and pulmonary cryptococcosis coexisting in an HTLV-1 seropositive patient. PMID- 14761149 TI - Physical and mental health survey of 20 patients with Sezary syndrome using the medical outcomes study 36-item short-form health survey. PMID- 14761150 TI - IgA pemphigus of the subcorneal pustular dermatosis type associated with monoclonal IgA gammopathy. PMID- 14761151 TI - Lupus erythematosus profundus involving the ear lobe. PMID- 14761152 TI - Dystrophic calcinosis cutis circumscripta. PMID- 14761153 TI - Mucosal involvement in childhood discoid lupus erythematosus. PMID- 14761154 TI - Toxic shock syndrome due to Staphylococcus aureus sepsis following diagnostic laparotomy for Hodgkin's disease. PMID- 14761155 TI - PICS VII and ENTICHS I: 7th Pediatric Interventional Cardiac Symposium & Emerging New Technologies in Congenital Heart Surgery, Orlando, Florida. PMID- 14761156 TI - Right coronary artery arising from the left ventricular outflow tract: a rare congenital anomaly of the coronary arteries. PMID- 14761157 TI - Past, present and future of combinatorial chemistry in Spain. PMID- 14761159 TI - Solid-phase synthesis of 2-substituted 4-amino-7-oxo-5,6,7,8-tetrahydropyrido[2,3 d]pyrimidines. AB - An efficient solid-phase synthesis of 2-substituted 4-aminopyrido[2,3 d]pyrimidines 15 is reported. The procedure started by solid supporting a p hydroxybenzaldehyde 8 to the Wang resin by using the Mitsunobu protocol. The resulting aldehyde 17 was treated with a substituted acid methyl malonate 10 to afford the corresponding alpha, beta-unsaturated ester 18, which was converted to the Michael adduct 21 by reaction with malononitrile. Cyclization of 21 with an amidine system 13 yielded the solid supported pyridopyrimidine 22, which afforded the corresponding 2-substituted 4-aminopyrido[2,3-d]pyrimidine 15 upon treatment with TFA:DCM. Compounds 15 present three diversity centers R1, R2 and R3. Having validated the chemistry on solid support, a 32-membered combinatorial library was obtained using this protocol. PMID- 14761158 TI - Exploring solid-phase approaches for the preparation of new beta-lactams from amino acids. AB - Two solid-phase approaches, involving the base-assisted intramolecular alkylation of N-chloroacetyl-Phe derivatives anchored to appropriate solid supports, were investigated for the preparation of novel beta-lactams. When a BAL-type strategy was used, the resin-bound azetidinones were easily formed, as established by MAS NMR, but final compounds could not be removed from the resin, unless a suitable two linkers system was used. In the second approach, in which the Phe residue is anchored to a Wang-type resin through the carboxylate group, the corresponding 1,4,4-trisubstituted 2-azetidinone was obtained in moderate to good yield and high purity. PMID- 14761160 TI - Immobilizing a single pybox ligand onto a library of solid supports. AB - Chiral pyridinebis(oxazoline) (pybox) ligands can be efficiently immobilized onto silica through position 4 of the pyridine ring. The crucial intermediate in this strategy is 4-chloropybox, which is prepared in good yield from chelidamic acid. 4-Chloropybox reacts with p-hydroxybenzaldehyde and p-aminophenol to give two intermediates (pybox-CHO and pybox-NH2) that allow to introduce the formyl and amino groups able to link to spacers with triethoxysilyl groups. The modified ligands and their ruthenium complexes are immobilized by grafting onto preformed silicas or, alternatively, the support is created by sol-gel synthesis using the functionalized chiral ligand as a silica monomer. In this way it is possible to create a library where the variation involves the support rather than the catalyst. The aim of this approach is to study the influence of different parameters, such as the textural properties of the support and the immobilization method, on the functionalization and catalytic performance. Some of the immobilized complexes are compared as heterogeneous catalysts in the cyclopropanation reaction of styrene with ethyl diazoacetate. PMID- 14761161 TI - Virtual generation of agents against Mycobacterium tuberculosis. A QSAR study. AB - A QSAR approach based on the use of various topological indices as new theoretical molecular descriptors was applied to the study of a set of 64 anti tuberculosis agents involving the substituted benzoxazines and phenylquinazolines. In order to evaluate the reliability of the proposed linear QSAR model, several statistical tests were proposed. The resulting model was subsequently applied to a wider virtual molecular library, which, together with the original set of 64 molecules with known activities contained another 512 molecules for which the predictions were made. Based on this prediction some new structures were proposed as especially promising candidates for active anti tuberculotic drugs. PMID- 14761162 TI - Analysis of selection methodologies for combinatorial library design. AB - We have implemented and adapted in Pralins (Program for Rational Analysis of Libraries in silico), the most popular sparse (cherry picking) and full array (sublibrary) selection algorithms: hierarchical clustering, k-means clustering, Optimum Binning, Jarvis Patrick, Pral-SE (partitioning techniques) and MaxSum, MaxMin, MaxMin averaged, DN2, CTD (distance-based methods). We have validated the program with an already synthesized three-component combinatorial library of FXR partial agonists characterized by standard computational chemistry descriptors as case study. This has let us analyze the goodness of both the partitioning techniques for space division and all the selection methodologies with respect to representativity in terms of population and space coverage for different selection sizes. Within the chemical space analyzed, both hierarchical clustering and Optimum Binning division strategies are found to be the most advantageous reference space divisions to be used in the subsequent population and space coverage studies. Complete hierarchical clustering appears also to be the preferred selection methodology for both sparse and full array problems. The full array restriction fulfillment can easily be overcome by convenient optimization algorithms that allow optimal reagent selection preserving > 90% of the population coverage. PMID- 14761163 TI - Use of alignment-free molecular descriptors in diversity analysis and optimal sampling of molecular libraries. AB - The selection of a sample of diverse compounds is a common strategy for exploring large molecular libraries. However, the success of such approach depends on the selection of relevant molecular descriptors and the use of appropriate sampling methods. In the context of pharmaceutical research, the molecular descriptors should be based on physicochemical properties related with the pharmacological behaviour of the compounds. In this sense, the alignment-free GRIND and VolSurf molecular descriptors are promising candidates since they have been successfully used in the modelling of both pharmacodynamic and pharmacokinetic properties of drugs. This work describes the use of such descriptors in the diversity sampling of a library of primary amines and compares the results with those obtained in a previous study that used quantum-mechanical descriptors. As in the previous work, principal component (PC) analysis was applied to reduce the dimensionality and remove redundant information of the original descriptors, and the compounds were sampled on the basis of k-means clustering on the space of the selected PCs. The results of the present study show that VolSurf and GRIND provide similar quality sampling regarding global features of the molecules such as hydrophilicity, however the topology of the compounds is considered differently. The similarity between particular compounds strongly depends on the original descriptors used. However all the sample selections done in the PC space after k-means clustering provide the same apparent diversity in comparison to the whole dataset. The results indicate that there is no best set of descriptors on a diversity basis. The selection of descriptors must be based on the drug features to be investigated. PMID- 14761164 TI - Combinatorial chemistry of beta-hairpins. AB - Combinatorial chemistry is expanding rapidly both in terms of chemistry development and application to the synthesis of compound libraries for lead discovery and optimization. Combinatorial technologies continue evolving and developing, in fact they are being used as basic research tools in different fields that include peptide/protein folding. This review examines the use of combinatorial chemistry in the design of peptides and protein domains that adopt beta-sheet conformations. In particular, the use of conformationally restricted peptide libraries has allowed the identification of linear peptides that are folded in a beta-hairpin structure in plain aqueous solutions. PMID- 14761165 TI - Use of combinatorial chemistry to develop photocurable thermoplastic polyurethane elastomers (TPUs). AB - Combinatorial chemistry was used to develop photocurable thermoplastic polyurethane elastomers through the incorporation of photoreactive diacetylene diols as chain extenders. The methodology applied allowed, in 36 experiments, the choice of the best compromise between mechanical properties and lack of colour. The combinations chosen were scaled up and their properties were evaluated in terms of mechanical properties. The combinatorial approach reduced the estimated time tenfold in developing such type of materials. PMID- 14761167 TI - A useful source for the drug-dry mouth relationship. PMID- 14761166 TI - 1H NMR spectroscopy with internal and external standards for the quantification of libraries. AB - A convenient and easy method based on 1H NMR spectroscopy with both external and internal standards is described for the quantification of members of libraries. PMID- 14761168 TI - Developing a pediatric oral health therapist to help address oral health disparities among children. AB - Oral Health in America: A Report of the Surgeon General documented the profound and significant disparities that exist in the oral health of children in the United States. Recently, the country has been issued a National Call to Action to Promote Oral Health, under the leadership of the Office of the Surgeon General. Among the significant factors contributing to the disparities problem is the access to oral health care by disadvantaged populations. There are inadequate numbers of dentists able and willing to treat children, particularly poor and minority children. In the early part of the twentieth century, New Zealand faced a significant problem with oral disease among its children and introduced a School Dental Service staffed by allied dental professionals, known as "school dental nurses," who had received two years training in caring for the teeth of children. A number of other countries have since adopted this model. This article reviews attempts to develop a comparable approach in the United States. Furthermore, it justifies and advocates the development of pediatric oral health therapists in the United States as a means of addressing the disparities problem that exists in this nation. These pediatric oral health therapists would be trained in a two-year program to provide dental care services to children. The article concludes by asserting that such action is a practical and cost-effective way for dentistry to fulfill its professional obligation to care for the oral health of all children, thus ensuring justice in oral health for America's children. PMID- 14761169 TI - General dentists' perceptions of educational and treatment issues affecting access to care for children with special health care needs. AB - This study analyzed a data subset of a national survey of general dentists conducted in 2001 to determine their overall care of children with special health care needs (CSHCN). In the survey, dentists were asked to respond to questions in the following areas: did they provide care for CSHCN (children with cerebral palsy, mental retardation, and those who are medically compromised); what were their perceptions of the training they received in dental school related to CSHCN; what was their interest in additional training for CSHCN; and what factors influenced their willingness to provide care for CSHCN? Only about 10 percent see CSHCN often or very often, and only one in four respondents had hands-on experience with these patients in dental school. Postgraduate education in general practice or advanced general dentistry residency had no effect on willingness to care for CSHCN. Older dentists, those accepting Medicaid for all children, and those practicing in small communities were more likely to see CSHCN. Dentists willing to see CSHCN also were more likely to perform procedures associated with special needs and underserved child populations including pharmacologic management and stainless steel crowns. Dentists with hands-on educational experiences in dental schools with CSHCN were less likely to consider such factors as level of disability and patient behavior as obstacles to care and were more likely to desire additional education in care of CSHCN. PMID- 14761170 TI - Comparisons of National Board Part II and NERB's written examination for outcomes and redundancy. AB - The purpose of this study was to compare students' in-school academic performance with their scores on the North East Regional Board (NERB) of Dental Examiners' Dental Simulated Clinical Examination (DSCE; written) and the written examination given by the National Board of Dental Examiners, Part II (NBDE Part II). A second purpose was to evaluate whether requiring the DSCE in addition to the NBDE II is of benefit in assessing the qualifications of candidates for licensure. Results on the first attempt at passing the two examinations were compared for 836 graduates (1994-2002) of one dental school. Mean scores on the examinations and mean grade point average (GPA) of those who passed the two examinations did not differ. Mean GPAs of those who failed the two examinations also were not significantly different. Scores on the two exams and GPA significantly correlated with each other. Correlation coefficients were higher between GPA and scores on the NBDE II than between GPA and scores on the DSCE. The failure rates on first attempts were 6 percent on the DSCE and 7 percent on the NBDE II. The results of this study suggest that the DSCE does not materially assist in the licensure decision for candidates who passed the NBDE Part II, as the results on the two exams are very similar. PMID- 14761172 TI - Preserving the pipeline: a model dental curriculum for research non-intensive institutions. AB - Current advanced degree and research training programs no longer attract adequate numbers of students. The present system of dental education severely limits the appeal of these programs due to overcrowded curricula and clinical components that operate in an environment segregated from the academic/research enterprise. To make research-oriented education/training programs more accessible and increase the number of interested students, the culture of dental schools and dental education must change. Programs for future dental researchers and academicians must be supported by curricula that foster an appreciation of research/discovery, an interest in academic/research careers, and the application of biomedical/clinical advances to practice. The Marquette University School of Dentistry has designed a comprehensive new curriculum that supports student research and scholarly activity throughout all four years of dental education. The curriculum minimizes discipline-based courses and is structured into interdisciplinary content tracks that integrate biomedical, behavioral, and clinical sciences while emphasizing the application of science to patient care. A specific research/scholarly track represents dedicated didactic time that exposes all dental students to material not traditionally included in dental curricula. This track includes mentored research/scholarly experiences at local and national sites that are individually structured for each student. Customized student schedules facilitate participation in these experiences without hardship or penalty. This curriculum structure may serve as a model for research non intensive institutions seeking to increase student interest in academic and research careers. PMID- 14761171 TI - Outcomes assessment related to professional growth and achievements of baccalaureate dental hygiene graduates. AB - The goals of the entry-level baccalaureate dental hygiene (DH) program at the University of California, San Francisco, include the preparation of scholars who will pursue additional study and research and the development of DH educators and leaders in the field of DH. The objective of this study was to evaluate whether the goals of the program were being achieved by assessing the professional growth and achievements of the graduates. Questionnaires were mailed to graduates of classes from 1960 to 2000, and the response rate was 66 percent (N = 468). Results indicated the following: 21 percent of the respondents completed a graduate or professional program, with most degrees being master's related to education (53 percent) and dental degrees (28 percent); 20 percent have held a faculty position in a DH educational program, with 26 percent holding leadership positions; 60 percent have been regular participants in DH professional organizations, with 36 percent of them holding leadership positions; and 61 percent have been active in a community organization, with 60 percent of them holding leadership positions. This outcomes assessment indicates that many UCSF DH graduates have become scholars, educators, and leaders in DH, and thus, the current program is successful at achieving its educational goals. Future studies should evaluate graduates of other DH programs on similar outcomes measures. PMID- 14761173 TI - Evaluation of a fissure sealant program as part of community-based teaching and training. AB - Since 1995 the Department of Community Dentistry of the University of Pretoria has been involved in the rendering of mobile primary oral health care services to children in the Hammanskraal area of Gauteng, South Africa, as part of their students' community-based training. Mokonyama Primary School was identified as the first school where a primary oral health care service could be rendered. The objective of this study was to evaluate the impact (outcomes) of a fissure sealant program on the dentition status of the school children. Seven years after the implementation of the program, the dentition status of children at Mokonyama was compared with that of a comparable group of children from the same area who were not exposed to the program. The results showed that the decayed, missing, and filled teeth in the primary dentition (dmft) in the six-year-old group in Mokonyama (1.74) did not differ significantly from the dmft (1.43) of the control group (p = 0.49). The decayed, missing, and filled teeth in the permanent dentition (DMFT) of 0.59 for the fifteen-year-old group in Mokonyama, however, differed significantly (p = 0.0001) from the DMFT of the control group (2.38). Fifteen-year-old children in Mokonyama had 75.2 percent fewer caries than their counterparts in the control group. PMID- 14761174 TI - The work covenant: a management strategy for department chairs and program directors. AB - Learning contracts have been used in adult education to ensure a positive and successful learning process. By means of a learning contract, the instructor can support the learner in a manner that best suits the learner's individual requirements. Like the learning environment, the workplace finds individuals with varying skill sets, areas of interest, and degrees of motivation. Understanding and applying the skills of the worker to appropriate tasks and interests have important ramifications for how workers perceive their jobs and the ultimate effectiveness of the organization. Studies show that the quality of the work environment is critical for the job satisfaction and retention of employees in the public and private sector as well as for the dental academician. Recent literature on faculty development has focused on adapting business-based motivation, mentoring, and performance counseling strategies to the academic setting. By leading and managing faculty development through the use of such strategies, the quality of the work environment can be improved. This article briefly summarizes the commonality between retention strategies and learning contracts within a human resource context and provides a model document that can be used by chairs and program directors to organize and focus their faculty development efforts. PMID- 14761175 TI - Application of rapid prototyping to operative dentistry curriculum. AB - Computers are becoming an integral part of dental education and dental practice. Rapid prototyping is a technique for the production of solid objects from computer models. The variety of rapid prototyping devices and materials that can be processed is increasing all the time. In this article, the authors present a discriminative learning device and associated exercises for developing visual recognition skills and examine the potential for the application of rapid prototyping to the operative dentistry curriculum. Other rapid prototyping technologies as applied in dentistry are also discussed. PMID- 14761176 TI - Problem-based learning and research at the Harvard School of Dental Medicine: a ten-year follow-up. AB - Dental students at the Harvard School of Dental Medicine (HSDM) are required to pursue and complete research projects in order to obtain the doctor of dental medicine (D.M.D.) degree. The purpose of this study was to evaluate the effect of curriculum changes on a set of measurable outcomes related to research pursuits. This study was designed as a retrospective analysis of outcomes data from the period 1992-2002. The predictor variable was program format (non-PBL vs. PBL); the outcome variables were the percentage of students listed as authors on an IADR/AADR abstract during their tenure at HSDM and the percentage of students listed as first and/or presenting authors. Univariate statistics were computed for each class, and independent samples t-tests were used to compare the study groups with regard to our outcomes. For the majority of our outcomes, there was no statistically significant difference between the PBL and non-PBL groups, possibly due to sample size limitations. While the implementation of PBL at HSDM has not universally increased student productivity in research, it has not had an adverse effect either. It is possible that students in the PBL curriculum are nurturing other interests, such as community service, while retaining productivity in research. PMID- 14761177 TI - Effect of grade point average and enrollment in a dental hygiene National Board review course on student performance on the National Board Examination. AB - Passing the National Board Dental Hygiene Examination is a requirement for licensure in all but one state. There are a number of preparation courses for the examination sponsored by corporations and dental hygiene programs. The purpose of this study was to determine if taking a board review course significantly affected student performance on the board examination. Students from the last six dental hygiene classes at Baylor College of Dentistry (n = 168) were divided into two groups depending on whether they took a particular review course. Mean entering college grade point averages (GPA), exiting dental hygiene program GPAs, and National Board scores were compared for the two groups using a t-test for independent samples (p < 0.05). No significant differences were found between the two groups for entering GPA and National Board scores. Exiting GPAs, however, were slightly higher for those not taking the course compared to those taking the course. In addition, a strong correlation (0.71, Pearson Correlation) was found between exiting GPA and National Board score. Exiting GPA was found to be a strong predictor of National Board performance. These results do not appear to support this program's participation in an external preparation course as a means of increasing students' performance on the National Board Dental Hygiene Examination. PMID- 14761178 TI - Perceived competency at graduation among dental alumni of the University of the West Indies. AB - The aims of this study were to describe levels of self-rated competency of dental graduates from the University of the West Indies (UWI) and to investigate relationships with gender and the effect of curriculum change. A thirty-two item self-reported postal questionnaire was sent to UWI dental alumni (1994-2002). The questionnaire included twenty-eight competencies that could be rated on a 5-point scale: 1 (not at all competent) to 5 (very competent). Overall preparedness for practice could also be rated from 1 (not at all prepared) to 5 (very prepared). The response rate was 77.4 percent, with a mean age of 29.3 years. Items with the highest mean scores were taking an adequate medical history (4.49), recognizing and treating dental caries (4.46), oral examination (4.36), and giving dental health education (4.35). Those with the lowest rating were designing and undertaking clinical research (2.29), dealing with practice management issues (2.52), designing and delivering crown and bridge work (3.33), and recognizing pathologic occlusions (3.33). Overall preparedness for practice was rated as 3.27. Female graduates rated four competencies significantly higher than males. Graduates exposed to the new curriculum perceived greater overall preparedness for general dental practice, suggesting the change to a competency-based curriculum was effective. PMID- 14761179 TI - Survey of Dental Student Financial Assistance, 2001-02. AB - The American Dental Education Association's 2001-02 Survey of Dental Student Financial Assistance obtained data by which to report, in aggregate and by type of school, the amount of financial assistance being received by dental students, in the form of loans, grants and scholarships, and work-study programs. Over 90 percent of the dental students received financial assistance through one or more federal, state, and/or school source. The average amount of assistance per student was dollar 35,100, ranging from an average of dollar 27,700 at public dental schools to dollar 51,100 at private dental schools. Loan programs accounted for almost 88 percent of all financial assistance; grants and scholarships, for 12 percent; work-study programs, for 0.2 percent. Overall, financial assistance exceeded average tuition and fees by 102 percent. With such levels of reliance on financial assistance, it remains imperative that students, even at the undergraduate level, receive the counseling, monitoring, and advice that will help them judiciously seek and manage appropriate types and amounts of financial assistance as they obtain their dental education. PMID- 14761180 TI - Inhibitors of serine proteases as potential therapeutic agents: the road from thrombin to tryptase to cathepsin G. PMID- 14761181 TI - Synthesis of a 5-methylindolyl-containing macrocycle that displays ultrapotent Grb2 SH2 domain-binding affinity. AB - The growth factor receptor-bound protein 2 (Grb2) is an SH2 domain-containing docking module that represents an attractive target for anticancer therapeutic intervention. Here, a ring-closing metathesis approach is utilized to synthesize a 5-methylindolyl-containing tetrapeptide mimetic (6) that exhibits unprecedented in vitro Grb2 SH2 domain-binding affinity (K(d) = 93 pM). Key to the preparation of 6 is the enantioselective synthesis of (2S)-2-(3-(5-methylindolyl)methyl)pent 4-enylamine (12) as one of two ring-closing segments. PMID- 14761183 TI - Virtual screening with flexible docking and COMBINE-based models. Application to a series of factor Xa inhibitors. AB - A two-step, fully automatic virtual screening procedure consisting of flexible docking followed by activity prediction by COMparative BINding Energy (COMBINE) analysis is presented. This novel approach has been successfully applied, as an example with medicinal chemistry interest, to a recently reported series of 133 factor Xa (fXa)(1) inhibitors whose activities encompass 4 orders of magnitude. The docking algorithm is linked to the COMBINE analysis program and used to derive independent regression models of the 133 inhibitors docked within three different fXa structures (PDB entries 1fjs, 1f0r, and 1xka), so as to explore the effect of receptor conformation on the overall results. Reliable docking conformations and predictive regression models requiring eight latent variables could be derived for two of the fXa structures, with the best model achieving a Q(2) of 0.63 and a standard deviation of errors of prediction (SDEP) of 0.51 (leave-one-out). The two-step procedure was then employed to screen a designed virtual library of 112 ligands, containing both active and inactive compounds. While docking energies alone could show a good performance for selecting hits, including structurally diverse ones, inclusion of COMBINE analysis regression models provided improved rankings for the identification of structurally related molecules in external sets. In our best case, a recognition rate of approximately 80% of known binders at approximately 15% false positives rate was achieved, corresponding to an enrichment factor of approximately 450% over random. PMID- 14761182 TI - In vitro structure-activity relationship and in vivo studies for a novel class of cyclooxygenase-2 inhibitors: 5-aryl-2,2-dialkyl-4-phenyl-3(2H)furanone derivatives. AB - 5-Aryl-2,2-dialkyl-4-phenyl-3(2H)furanone derivatives were studied as a novel class of selective cyclooxygenase-2 inhibitors with regard to synthesis, in vitro SAR, antiinflammatory activities, pharmacokinetic considerations, and gastric safety. 1f, a representative compound for methyl sulfone derivatives, showed a COX-2 IC(50) comparable to that of rofecoxib. In case of 20b, a representative compound for sulfonamide derivatives, a potent antiinflammatory ED(50) of 0.1 mg kg(-1) day(-1) was observed against adjuvant-induced arthritis by a preventive model, positioning 20b as one of the most potent COX-2 inhibitors ever reported. Furthermore, 20b showed strong analgesic activity as indicated by its ED(50) of 0.25 mg/kg against carrageenan-induced thermal hyperalgesia in the Sprague-Dawley rat. 3(2H)Furanone derivatives showed due gastric safety profiles as selective COX-2 inhibitors upon 7-day repeat dosing. A highly potent COX-2 inhibitor of the 3(2H)furanone scaffold could be considered suitable for a future generation COX-2 selective arthritis medication with improved safety profiles. PMID- 14761184 TI - Prediction of HIV-1 integrase/viral DNA interactions in the catalytic domain by fast molecular docking. AB - This study details the separate analyses of binding specificity of HIV-1 integrase (IN) and viral B-DNA forms through ligand-receptor docking studies by means of a fast molecular docking method. The application of solvated electrostatics with the University of Houston Brownian Dynamics Program (UHBD) and configurational sampling by the Daughter of Turnip (DOT) docking program resulted in the computation of energies of more than 113 billion configurations for each ligand-receptor docking study, a procedure considered computationally intractable a few years ago. A specific binding pattern of viral DNA to the IN catalytic domain region has been predicted as a result of these calculations. In a representative docked configuration, we observe the 3'-hydroxyl of the conserved deoxyadenosine to be close to one of the two divalent metal ions that are necessary for catalysis. A superimposition of our energy-minimized docked complex on representative structures from a molecular dynamics (MD) simulation of a crystallographically resolved IN/inhibitor complex revealed an overlap of viral DNA with the inhibitor, indicating that the bound inhibitor might operate by blocking substrate binding. The DOT docking calculation also identified a second, adjacent DNA-binding site, which we believe is the nonspecific host DNA binding site. The binding pattern predicted by DOT complements previous electrostatics, MD simulation, photo-cross-linking, and mutagenesis studies and also provides a further refinement of the IN/viral DNA binding interaction as a basis for new structure-based design efforts. PMID- 14761185 TI - Explorations of peptide and oligonucleotide binding sites of tyrosyl-DNA phosphodiesterase using vanadate complexes. AB - Tyrosyl-DNA phosphodiesterase (Tdp1) catalyzes the hydrolysis of a phosphodiester bond between a tyrosine residue and a DNA 3' phosphate and functions as a DNA repair enzyme that cleaves stalled topoisomerase I-DNA complexes. We previously determined a procedure to crystallize a quaternary complex containing Tdp1, vanadate, a DNA oligonucleotide, and a tyrosine-containing peptide that mimics the transition state for hydrolysis of the Tdp1 substrate. Here, the ability of vanadate to accept a variety of different ligands is exploited to produce several different quaternary complexes with a variety of oligonucleotides, and peptides or a tyrosine analogue, in efforts to explore the binding properties of the Tdp1 DNA and peptide binding clefts. Eight crystal structures of Tdp1 with vanadate, oligonucleotides, and peptides or peptide analogues were determined. These structures demonstrated that Tdp1 is able to bind substituents with limited sequence variation in the polypeptide moiety and also bind oligonucleotides with sequence variation at the 3' end. Additionally, the tyrosine analogue octopamine can replace topoisomerase I derived peptides as the apical ligand to vanadate. The versatility of this system suggests that the formation of quaternary complexes around vanadate could be adapted to become a useful method for structure-based inhibitor design and has the potential to be generally applicable to other enzymes that perform chemistry on phosphate esters. PMID- 14761186 TI - Undersulfated and glycol-split heparins endowed with antiangiogenic activity. AB - Tumor neovascularization (angiogenesis) is regarded as a promising target for anticancer drugs. Heparin binds to fibroblast growth factor-2 (FGF2) and promotes the formation of ternary complexes with endothelial cell surface receptors, inducing an angiogenic response. As a novel strategy to generate antiangiogenic substances exploiting binding to FGF2 while preventing FGF receptor (FGFR) activation, sulfation gaps were generated along the heparin chains by controlled alkali-catalyzed removal of sulfate groups of iduronic acid 2-O-sulfate residues, giving rise to the corresponding epoxide derivatives. A new class of heparin derivatives was then obtained by opening the epoxide rings followed by oxidative glycol-splitting of the newly formed (and the preexisting) nonsulfated uronic acid residues. In vitro these heparin derivatives prevent the formation of FGFR/FGF2/heparan sulfate proteoglycan ternary complexes and inhibit FGF2 stimulated endothelial cell proliferation. They exert an antiangiogenic activity in the chick embryo chorioallantoic membrane assay, where the parent heparin is inactive. Low and very low molecular weight derivatives of a prototype compound, as well as its glycine and taurine derivatives obtained by reductive amination of glycol-split residues, retained the angiostatic activity. A significant relationship was found between the extent of glycol-splitting and the FGF2 antagonist/angiostatic activities of these heparin derivatives. Molecular dynamics calculations support the assumption that glycol-split residues act as flexible joints that, while favoring 1:1 binding to FGF2, disrupt the linearity of heparin chains necessary for formation of active complexes with FGFRs. PMID- 14761187 TI - Antitumor agents. 3. Design, synthesis, and biological evaluation of new pyridoisoquinolindione and dihydrothienoquinolindione derivatives with potent cytotoxic activity. AB - New antiproliferative compounds, the 1-aryl-3-ethoxycarbonyl-pyrido[2,3 g]isoquinolin-5,10-diones (PIQDs, 1-7), were designed on the basis of a molecular model obtained by aligning the common quinolinquinone substructure of 5H pyrido[3,2-a]phenoxazin-5-one (PPH) and some known anticancer agents. A Diels Alder reaction between quinolin-5,8-dione (QD) and a 2-azadiene, formed by demolition of 2-aryl-1,3-thiazolidine ethyl esters (T compounds), was used to produce 1-7 and the isomeric 1-aryl-3-ethoxycarbonylpyrido[3,2-g]isoquinolin-5,10 diones (8-14). Two other compounds, the 3-amino-3-ethoxycarbonyldihydrothieno[2,3 g]quinolin-4,9-dione (15) and the 3-amino-3-ethoxycarbonyldihydrothieno[3,2 g]quinolin-4,9-dione (16), arising from a 1,4 Michael reaction of QD with a thiolate species formed by opening of T compounds, were recovered from the reaction mixture. The antiproliferative activity of 1-16 was evaluated against representative human liquid and solid neoplastic cell lines. The IC(50) of these compounds had median values in the range 2.00-0.01 microM, with 2-4 and 15 exhibiting significantly higher in vitro cytotoxic activity. Compound 2, also evaluated against KB subclones (KB(MDR), KB(7D), and KB(V20C)), was shown to be scarcely subject to the MDR1/P-glycoprotein drug efflux pump responsible for drug resistance. The noncovalent DNA-binding properties of PIQDs were examined using UV-vis and (1)H NMR spectroscopy experiments. Accordingly, these compounds were confirmed to have an ability to intercalate into double-stranded DNA by topoisomerase I superhelix unwinding assay. Interesting structure-activity relationships were found. Three important features seem to contribute to the cytotoxic activity of these anticancer ligands: (i) the DNA intercalating capability of the three-cyclic quinonic system, typical of this class of compounds, (ii) the position of the pendant phenyl ring that, according to the superimposition model, must occupy the same area of the corresponding benzo-fused ring A of PPH, and (iii) the effect of electron-withdrawing substituents on the phenyl ring, which can contribute improving the pi-pi stacking interactions between ligand and DNA base pairs. Besides, a mechanism of action suspected to involve topoisomerases could be hypothesized to interpret the antiproliferative activity of the thienoquinolindione 15, which can be regarded as a cyclic cysteine derivative. PMID- 14761188 TI - A selective AT2 receptor ligand with a gamma-turn-like mimetic replacing the amino acid residues 4-5 of angiotensin II. AB - Three angiotensin II (Ang II) analogues encompassing a benzodiazepine-based gamma turn-like scaffold have been synthesized. Evaluation of the compounds in a radioligand binding assay showed that they had no affinity to the rat liver AT(1) receptor. However, one of the compounds displayed considerable affinity to the pig uterus AT(2) receptor (K(i) = 3.0 nM) while the other two lacked affinity to this receptor. It was hypothesized that the reason for the inactivity of one of these analogues to the AT(2) receptor was that the guanidino group of the Arg(2) residue and/or the N-terminal end of the pseudopeptide could not interact optimally with the receptor. To investigate this hypothesis, a conformational analysis was performed and a comparison was carried out with the monocyclic methylenedithioether analogue cyclo(S-CH(2)-S)[Cys(3,5)]Ang II which is known to bind with high affinity to the AT(2) receptor (K(i) = 0.62 nM). This comparison showed that, in the compounds with high AT(2) receptor affinity, the guanidino group of the Arg(2) residue and the N-terminal end could access common regions of space that were not accessible to the inactive compound. To examine the importance of the guanidino group for binding, the Arg side chain was removed by substituting Arg(2) for Ala(2) in the analogue having the high affinity. This analogue lacked affinity to AT(2) receptors, which supports the role of the guanidino group in receptor binding. PMID- 14761189 TI - Discovery of a new class of anilinoquinazoline inhibitors with high affinity and specificity for the tyrosine kinase domain of c-Src. AB - Deregulated activity of the nonreceptor tyrosine kinase c-Src is believed to result in signal transduction, cytoskeletal and adhesion changes, ultimately promoting a tumor-invasive phenotype. We report here the discovery of a new class of anilinoquinazoline inhibitors with high affinity and specificity for the tyrosine kinase domain of the c-Src enzyme. Special attention was directed toward finding inhibitors selective against KDR tyrosine kinase in order to ensure that the in vivo profile of a specific Src inhibitor could be determined. The 4 aminobenzodioxole quinazoline series gave compounds with excellent potency and selectivity. The most interesting compounds were evaluated in vivo and displayed good pharmacokinetics following oral dosing. Compounds such as the aminobenzodioxoles were shown to be potent inhibitors of tumor growth in a c-Src transformed 3T3 xenograft model in vivo, resulting in more than 90% growth inhibition at doses as low as 6 mg/kg po once daily. Src tyrosine kinase inhibitors such as these may provide a novel therapeutic modality for targeting cancer invasion and metastasis. PMID- 14761190 TI - Recognition of privileged structures by G-protein coupled receptors. AB - Privileged structures are ligand substructures that are widely used to generate high-affinity ligands for more than one type of receptor. To explain this, we surmised that there must be some common feature in the target proteins. For a set of class A GPCRs, we found a good correlation between conservation patterns of residues in the ligand binding pocket and the privileged structure fragments in class A GPCR ligands. A major part of interior surface of the common ligand binding pocket of class A receptors, identified in many GPCRs, is lined with variable residues that are responsible for selectivity in ligand recognition, while other regions, typically located deeper into the binding pocket, are more conserved and retain a predominantly hydrophobic and aromatic character. The latter is reflected in the chemical nature of most GPCR privileged structures and is proposed to be the common feature that is recognized by the privileged structures. Further, we find that this subpocket is conserved even in distant orthologs within the class A family. Three pairs of ligands recognizing widely different receptor types were docked into receptor models of their target receptors utilizing available structure- activity relationships and mutagenesis data. For each pair of ligands, the ligand-receptor complexes reveal that the nature of the privileged structure binding pocket is conserved between the two complexes, in support of our hypothesis. Only part of the privileged structures can be accommodated within the conserved subpocket. Some contacts are established between the privileged structure and the nonconserved parts of the binding pocket. This implies that any one particular privileged structure can target only a subset of receptors, those complementary to the full privileged structure. Our hypothesis leads to a valuable novelty in that ligand libraries can be designed without any foreknowledge of the structure of the endogenous ligand, which in turn means that even orphan receptors can in principle now be addressed as potential drug targets. PMID- 14761191 TI - 5-Fluorinated L-lysine analogues as selective induced nitric oxide synthase inhibitors. AB - 5(S)-Fluoro-N6-(iminoethyl)-l-lysine (14), an analogue of the potent, selective induced nitric oxide synthase (iNOS) inhibitor iminoethyl-l-lysine (1), was synthesized and found to be a selective iNOS inhibitor. PMID- 14761192 TI - Conformer- and alignment-independent model for predicting structurally diverse competitive CYP2C9 inhibitors. AB - A conformer- and alignment-independent three-dimensional structure-activity relationship (3D-QSAR) model has been derived that is based on flexible molecular interaction fields calculated in GRID and the subsequent description of these fields by use of alignment-independent descriptors derived in ALMOND. The training set consisted of 22 diverse and flexible competitive inhibitors of the drug-metabolizing enzyme CYP2C9 and generated a model with r(2) of 0.81 and q(2) of 0.62. The predicitive capacity of the model was externally evaluated with a test set of 12 competitive inhibitors and 11 out of 12 were predicted within 0.5 log unit. The most relevant points of interaction in the model correlated well to the amino acids involved in CYP2C9-substrate/inhibitor binding in the active site of a CYP2C9 homology model, further validating the mechanistic sense of our model. This approach offers the possibility to derive predicitve 3D-QSAR models without the need for an alignment rule for chemically diverse ligands and in the absence of target protein crystal structure information. PMID- 14761193 TI - Effects of C2-alkylation, N-alkylation, and N,N'-dialkylation on the stability and estrogen receptor interaction of (4R,5S)/(4S,5R)-4,5-bis(4-hydroxyphenyl)-2 imidazolines. AB - (4R,5S)/(4S,5R)-4,5-Bis(4-hydroxyphenyl)-2-imidazolines bearing 2,2'-H (3a), 2,2' Cl (3b), 2,2',6-Cl (3c), and 2,2'-F (3d) substituents in the aromatic rings were C2-alkylated (5a-i), N-alkylated (7, 7a-c), and N,N'-dialkylated (9a-c). The synthesis started from the diastereomerically pure (1R,2S)/(1S,2R)-1,2-diamino 1,2-bis(4-methoxyphenyl)ethanes 1a-d, which were cyclized to the imidazolines 2a d and 4a-i with triethylorthoesters or iminoethers. Ether cleavage with BBr(3) yielded the (4R,5S)/(4S,5R)-4,5-bis(4-hydroxyphenyl)-2-imidazolines 3a-d and 5a i. The N-alkylation and N,N'-dialkylation of 2b, employed for obtaining 7a-c and 9a-c, were performed prior to the ether cleavage with alkyl iodine in dry THF. By use of HPLC, the influence of the substitution patterns in the aromatic rings and alkyl chains at the C2- or N-atoms on the hydrolysis rate of the imidazolines was studied under in vitro conditions. It appeared that only imidazolines with C2- or N-alkyl substituents show sufficient stability to interact as heterocycles with the estrogen receptor (ER). The resulting gene activation was monitored in a luciferase assay using ERalpha-positive MCF-7-2a breast cancer cells stably transfected with the plasmid ERE(wtc)luc. It is interesting to note that C2 alkylation led to a strong reduction or even a complete loss of activity whereas N-alkylation improved the estrogenic profile. The (4R,5S)/(4S,5R)-N-ethyl-4,5 bis(2-chloro-4-hydroxyphenyl)-2-imidazoline 7b has proven to be the most active compound in this structure-activity relationship study (EC(50) = 0.015 microM). PMID- 14761194 TI - Computer-aided design, synthesis, and anti-HIV-1 activity in vitro of 2 alkylamino-6-[1-(2,6-difluorophenyl)alkyl]-3,4-dihydro-5-alkylpyrimidin-4(3H) ones as novel potent non-nucleoside reverse transcriptase inhibitors, also active against the Y181C variant. AB - Dihydro-alkoxy-benzyl-oxopyrimidines (DABOs) are a family of potent NNRTIs developed in the past decade. Attempts to improve their potency and selectivity led to thio-DABOs (S-DABOs), DATNOs, and difluoro-thio-DABOs (F(2)-S-DABOs). More recently, we reported the synthesis and molecular modeling studies of a novel conformationally constrained subtype of the S-DABO series characterized by the presence of substituents on the methylene linkage connecting the pyrimidine ring to the aryl moiety (Mai, A., et al. J. Med. Chem. 2001, 44, 2544-2554). Now we report the computer-aided design, synthesis, and biological evaluation of four new DABO prototypes (5-alkyl-2-cyclopentylamino-6-[1-(2,6-difluorophenyl)alkyl] 3,4-dihydropyrimidin-4(3H)-ones, F(2)-NH-DABOs) in which the sulfur atom of the related F(2)-S-DABOs is replaced by an amino group. For these studies, we used as a reference model the cocrystallized MKC-442/RT complex. Docking studies with Autodock of the newly designed F(2)-NH-DABOs on the ligand-derived RT confirmed the findings previously described for the F(2)-S-DABOs. The F(2)-NH-DABO binding mode resembles that reported for F(2)-S-DABOs, with the difference that the NH moiety at the C-2 position represents a new anchor site for ligand/enzyme complexation. The predicted inhibition constant (K(i)) values by the internal scoring function of Autodock, and the predicted IC(50) values by the application of a VALIDATE II/HIV-RT model strongly suggested the synthesis of the designed amino-DABOs. F(2)-NH-DABOs were shown to be highly active in both anti-RT and anti-HIV biological assays with IC(50) and EC(50) comparable with that of the reference compound MKC-442. Interestingly, 2-cyclopentylamino-6-[1-(2,6 difluorophenyl)ethyl]-3,4-dihydro-5-methyl pyrimidin-4(3H)-one (9d) was active against the Y181C HIV-1 mutant strain at submicromolar concentration, with a resistance value similar to that of efavirenz, the last FDA-approved NNRTI for AIDS therapy, and 2-fold lower than that of its 2-cyclopentylthio counterpart 8d. The introduction in 9d of a new anchor point (pyrimidine C-2 NH group), with the formation of a new hydrogen bond with Lys101, could compensate for the lack of positive hydrophobic ligand/NNBP interactions due to the Tyr181 to Cys181 mutation. PMID- 14761195 TI - Structural basis for the synthesis of indirubins as potent and selective inhibitors of glycogen synthase kinase-3 and cyclin-dependent kinases. AB - Pharmacological inhibitors of glycogen synthase kinase-3 (GSK-3) and cyclin dependent kinases have a promising potential for applications against several neurodegenerative diseases such as Alzheimer's disease. Indirubins, a family of bis-indoles isolated from various natural sources, are potent inhibitors of several kinases, including GSK-3. Using the cocrystal structures of various indirubins with GSK-3beta, CDK2 and CDK5/p25, we have modeled the binding of indirubins within the ATP-binding pocket of these kinases. This modeling approach provided some insight into the molecular basis of indirubins' action and selectivity and allowed us to forecast some improvements of this family of bis indoles as kinase inhibitors. Predicted molecules, including 6-substituted and 5,6-disubstituted indirubins, were synthesized and evaluated as CDK and GSK-3 inhibitors. Control, kinase-inactive indirubins were obtained by introduction of a methyl substitution on N1. PMID- 14761196 TI - Ligand-based structural hypotheses for virtual screening. AB - The majority of drug targets for small molecule therapeutics are proteins whose three-dimensional structure is not known to sufficient resolution to permit structure-based design. All three-dimensional QSAR approaches have a requirement for some hypothesis of ligand conformation and alignment, and predictions of molecular activity critically depend on this ligand-based binding site hypothesis. The molecular similarity function used in the Surflex docking system, coupled with quantitative pressure to minimize overall molecular volume, forms an effective objective function for generating hypotheses of bioactive conformations of sets of small molecules binding to their cognate proteins. Results are presented, assessing utility of the method for ligands of the serotonin, histamine, muscarinic, and GABA(A) receptors. The Surflex similarity module (Surflex-Sim) was able, in each case, to distinguish true ligands from random compounds using models constructed from just two or three known ligands. True positive rates of 60% were achieved with false positive rates of 0-3%; the theoretical enrichment rates were over 150-fold compared with random screening. The methods are practically applicable for rational design of ligands and for high-throughput virtual screening and offer competitive performance to many structure-based docking algorithms. PMID- 14761197 TI - Synthesis, SAR, crystal structure, and biological evaluation of benzoquinoliziniums as activators of wild-type and mutant cystic fibrosis transmembrane conductance regulator channels. AB - Chloride channels play important roles in homeostasis and regulate cell volume, transepithelial transport, and electrical excitability. Despite recent progress made in the genetic and molecular aspect of chloride channels, their pharmacology is still poorly understood. The cystic fibrosis transmembrane conductance regulator (CFTR) is a cAMP-regulated epithelial chloride channel for which mutations cause cystic fibrosis. Here we have synthesized benzo[c]quinolizinium and benzo[f]indolo[2,3-a]quinolizinium salts (MPB) and performed a SAR to identify the structural basis for activation of the CFTR chloride channel. Synthesized compounds were evaluated on wild-type CFTR and on CFTR having the glycine-to-aspartic acid missense mutation at codon 551 (G551D-CFTR), using a robot and cell-based assay. The presence of an hydroxyl group at position 6 of the benzo[c]quinolizinium skeleton associated with a chlorine atom at position 10 or 7 and an alkyl chain at position 5 determined the highest activity. The most potent product is 5-butyl-7-chloro-6-hydroxybenzo[c]quinolizinium chloride (8u, MPB-104). 8u is 100 times more potent than the parent compound 8a (MPB-07). PMID- 14761198 TI - Formation of water-soluble pincer silver(I)-carbene complexes: a novel antimicrobial agent. AB - Silver(I)-2,6-bis(ethanolimidazolemethyl)pyridine hydroxide (4a) and silver(I) 2,6-bis(propanolimidazolemethyl)pyridine hydroxide (4b) are water-soluble silver(I)-carbene complexes that were synthesized in high yield by reacting silver(I) oxide with N-substituted pincer ligands 3 (a = 2,6 bis(ethanolimidazoliummethyl)pyridine diiodide, b = 2,6 bis(propanolimidazoliummethylpyridine)pyridine dibromide). The X-ray crystal structure of 4a is a one-dimensional linear polymer, whereas the mass spectroscopy confirms a monomer in the gas phase. A change in the anion of 4a from a hydroxide to a hexafluorophosphate formed a silver(I)-carbene complex 4c that is dimeric in structure and insoluble in water. The bactericidal activities of the water-soluble silver(I)-carbene complexes were found to be improved over that of silver nitrate. PMID- 14761199 TI - Synthesis of mono- and bisdihydrodipyridopyrazines and assessment of their DNA binding and cytotoxic properties. AB - Aminoalkyl-substituted monomeric and dimeric dihydrodipyridopyrazines have been synthesized and evaluated as antitumor agents. Potent cytotoxic compounds were identified in both series. Biochemical and biophysical studies indicated that all these compounds strongly stabilized the duplex structure of DNA and some of them elicited a selectivity for GC-rich sequences. Sequence recognition by of the dimeric dihydrodipyridopyrazines is reminiscent of that of certain antitumor bisnaphthalimides. Compared to monomers, corresponding dimeric derivatives showed higher affinity for DNA. This property was attributed to a bisintercalative binding to DNA. This assumption was indirectly probed by electric linear dichroism and DNA relaxation experiments. DNA provides a bioreceptor for these dihydrodipyridopyrazine derivatives, but no poisoning of human topoisomerases I or II was detected. Most of the compounds efficiently inhibited the growth of L1210 murine leukemia cells and perturbed the cell cycle progression (with a G2/M block in most cases). A weak but noticeable in vivo antitumor activity was observed with one of the dimeric compounds. This studies identifies monomeric and dimeric dihydrodipyridopyrazines as a new class of DNA-targeted antitumor agents. PMID- 14761200 TI - Jatrophane diterpenes as modulators of multidrug resistance. Advances of structure-activity relationships and discovery of the potent lead pepluanin A. AB - From the whole plant of Euphorbia peplus L., five new diterpenes based on a jatrophane skeleton (pepluanins A-E, 1-5) were isolated, together with two known analogues (6 and 7), which served to divulge in detail the structure-activity relationships within this class of P-glycoprotein inhibitors. The results revealed the importance of substitutions on the medium-sized ring (carbons 8, 9, 14, and 15). In particular, the activity is collapsed by the presence of a free hydroxyl at C-8, while it increases with a carbonyl at C-14, an acetoxyl at C-9, and a free hydroxyl at C-15. The most potent compound of the series, pepluanin A, showed a very high activity for a jatrophane diterpene, outperforming cyclosporin A by a factor of at least 2 in the inhibition of Pgp-mediated daunomycin transport. PMID- 14761201 TI - Design, synthesis, and biological characterization of metabolically stable selective androgen receptor modulators. AB - A series of nonsteroidal ligands were synthesized as second-generation agonists for the androgen receptor (AR). These ligands were designed to eliminate metabolic sites identified in one of our first-generation AR agonists, which was inactive in vivo due to its rapid metabolism to inactive constituents. The binding affinity of these compounds was evaluated using AR isolated from rat ventral prostate. These second-generation compounds bound the AR in a high affinity and stereoselective manner, with K(i) values ranging from about 4 to 130 nM. The ability of these ligands to stimulate AR-mediated transcriptional activation was examined in cells transfected with the human AR and a hormone dependent luciferase reporter gene. Although some compounds were unable to stimulate AR-mediated transcription, several demonstrated activity similar to that of dihydrotestosterone (DHT, an endogenous steroidal ligand for the AR). We also evaluated the in vivo pharmacologic activity of selected compounds in castrated male rats. Three compounds were identified as selective androgen receptor modulators (SARMs), exhibiting significant anabolic activity while having only moderate to minimal androgenic activity in vivo. PMID- 14761202 TI - Synthesis of flexible sulfur-containing heteroarotinoids that induce apoptosis and reactive oxygen species with discrimination between malignant and benign cells. AB - Regulation of growth, differentiation, and apoptosis by synthetic retinoids can occur through mechanisms that are dependent and independent of their ability to bind and activate nuclear retinoic acid receptors. The objective of this study was to determine if increasing flexibility of the heteroarotinoid structure would affect the specificity of the synthetic retinoids for the receptors and for their regulation of cancerous and nonmalignant cells. Methods were developed to produce the first examples of heteroarotinoids 15a-15h, which contain urea and/or thiourea linking groups between two aryl rings. Substituents at the para position of the single phenyl ring were either an ester, a nitro group, or a sulfonamide group. Ovarian cancer cell lines Caov-3, OVCAR-3, SK-OV-3, UCI-101, and 222 were utilized, and the inhibitory prowess of the heteroarotinoids was referenced to that of 4-HPR (25). Similar to 4-HPR (25), the heteroarotinoids inhibited growth of all cell lines at micromolar concentrations. Although the heteroarotinoids did not activate retinoic acid receptors, the agents induced potent growth inhibition against the cancer cells with weak activity against normal and benign cells. The growth inhibition was associated with cell loss and induction of reactive oxygen species. PMID- 14761203 TI - Novel heteroarotinoids as potential antagonists of Mycobacterium bovis BCG. AB - A series of 15 heteroarotinoids has been prepared and evaluated for activity against Mycobacterium bovis BCG with the thiourea-containing isoxyl (7) (0.5 microg/mL) as the standard. 2,2,4-Trimethyl-2H-chromen-7-yl 4 (methoxycarbonyl)benzoate (8) displayed the most significant activity (2.0-4.0 microg/mL) in terms of the lowest concentration (microg/mL) (MIC, minimum inhibitory concentration) required to produce a 99% reduction in the number of colonies on a plate as compared to that system free of the agent at the same dilution of the culture suspension. Ethyl 4-[[N-(2,2,4,4-tetramethylchroman-6 yl)thiocarbamoyl]amino]benzoate (9) and [[(1E,3Z,5E)-1-aza-4-methyl-6-(1,2,2,4 tetramethyl(1,2-dihydroquinolyl))hexa-1,3,5-trienyl]amino]aminomethane-1-thione (10) exhibited activity at 5.0-10.0 and 10.0-20.0 microg/mL, respectively, while the other examples had MIC values of 20 microg/mL or greater. The inhibitory ability of 8 may occur via the inhibition of mycolic acid synthesis in a like manner as found with 7, but this requires further study. The heteroarotinoids are the first examples to exhibit inhibitory ability against the growth of Mycobacterium bovis BCB. PMID- 14761204 TI - Prediction of ligand binding affinity and orientation of xenoestrogens to the estrogen receptor by molecular dynamics simulations and the linear interaction energy method. AB - Exposure to environmental estrogens has been proposed as a risk factor for disruption of reproductive development and tumorigenesis of humans and wildlife (McLachlan, J. A.; Korach, K. S.; Newbold, R. R.; Degen, G. H. Diethylstilbestrol and other estrogens in the environment. Fundam. Appl. Toxicol. 1984, 4, 686-691). In recent years, many structurally diverse environmental compounds have been identified as estrogens. A reliable computational method for determining estrogen receptor (ER) binding affinity is of great value for the prediction of estrogenic activity of such compounds and their metabolites. In the presented study, a computational model was developed for prediction of binding affinities of ligands to the ERalpha isoform, using MD simulations in combination with the linear interaction energy (LIE) approach. The linear interaction energy approximation was first described by Aqvist et al. (Aqvist, J.; Medina, C.; Samuelsson, J. E. A new method for predicting binding affinity in computer-aided drug design. Protein Eng. 1994, 7, 385-391) and relies on the assumption that the binding free energy (DeltaG) depends linearly on changes in the van der Waals and electrostatic energy of the system. In the present study, MD simulations of ligands in the ERalpha ligand binding domain (LBD) (Shiau, A. K.; Barstad, D.; Loria, P. M.; Cheng, L.; Kushner, P. J.; Agard, D. A.; Greene, G. L. The structural basis of estrogen receptor/coactivator recognition and the antagonism of this interaction by tamoxifen. Cell 1998, 95, 927-937), as well as ligands free in water, were carried out using the Amber 6.0 force field (http://amber.scripps.edu/). Contrary to previous LIE methods, we took into account every possible orientation of the ligands in the LBD and weighted the contribution of each orientation to the total binding affinity according to a Boltzman distribution. The training set (n = 19) contained estradiol (E2), the synthetic estrogens diethylstilbestrol (DES) and 11beta-chloroethylestradiol (E2-Cl), 16alpha-hydroxy-E2 (estriol, EST), the phytoestrogens genistein (GEN), 8-prenylnaringenin (8PN), and zearalenon (ZEA), four derivatives of benz[a]antracene-3,9-diol, and eight estrogenic monohydroxylated PAH metabolites. We obtained an excellent linear correlation (r(2) = 0.94) between experimental (competitive ER binding assay) and calculated binding energies, with K(d) values ranging from 0.15 mM to 30 pM, a 5 000 000 fold difference in binding affinity. Subsequently, a test set (n = 12) was used to examine the predictive value of our model. This set consisted of the synthetic estrogen 5,11-cis-diethyl-5,6,11,12-tetrahydrochrysene-2,8-diol (THC), daidzein (DAI), equol (EQU) and apigenin (API), chlordecone (KEP), progesterone (PRG), several mono- and dihydroxylated PAH metabolites, and two brominated biphenyls. The predicted binding affinities of these estrogenic compounds were in very good agreement with the experimental values (average deviation of 0.61 +/- 0.4 kcal/mol). In conclusion, our LIE model provides a very good method for prediction of absolute ligand binding affinities, as well as binding orientation of ligands. PMID- 14761205 TI - Preparation, properties, reactions, and adenosine receptor affinities of sulfophenylxanthine nitrophenyl esters: toward the development of sulfonic acid prodrugs with peroral bioavailability. AB - Many currently known antagonists for P2 purinergic receptors are anionic molecules bearing one or several phenylsulfonate groups. Among the P1 (adenosine) receptor antagonists, the xanthine phenylsulfonates are a potent class of compounds. Due to their high acidity, phenylsulfonates are negatively charged at physiologic pH values and do not easily penetrate cell membranes. The present study was aimed at developing lipophilic, perorally bioavailable prodrugs of sulfonates by converting them into chemically stable nitrophenyl esters. Initial stability tests at different pH values using nitrophenyl tosylates as model compounds showed that m-nitrophenyl esters were stable over a wide pH range, while the ortho and para isomers were less stable under strongly acidic or basic conditions. A series of m- and p-nitrophenyl esters of p-sulfophenylxanthine derivatives were synthesized as model compounds. The target xanthine derivatives were obtained in high yields by condensation of the appropriate 5,6 diaminouracils with 4-(nitrophenoxysulfonyl)benzoic acids in the presence of a carbodiimide, followed by ring closure with polyphosphoric acid trimethylsilyl ester. The chemical and enzymatic stability of the m-nitrophenyl esters was investigated in vitro by means of capillary electrophoresis. High stability in aqueous solution, in artificial gastric acid, and in serum was observed. However, compound 5d, used as a prototypic xanthine m-nitrophenylsulfonate, was hydrolyzed by rat liver homogenate indicating an enzymatic pathway of hydrolysis. Thus, nitrophenyl esters of sulfonic acids have a potential as peroral prodrugs of drugs bearing a sulfonate group. The nitrophenyl esters of sulfophenylxanthines were additionally investigated for their adenosine receptor affinities. They showed high affinity at A(1), A(2A), and A(2B), but not at A(3) ARs. One of the most potent compounds was 1-propyl-8-[4-[[p-nitrophenoxy]sulfonyl]phenyl]xanthine (9d), a mixed A(1)/A(2B) antagonist (K(i)A(1) 3.6 nM, K(i)A(2B) 5.4 nM) selective versus the other subtypes. As a further result of this study, the m-nitrophenoxy group was found to be a suitable protecting group for sulfonates in organic synthesis due to its high lipophilicity and stability; it can be split off under strongly basic conditions. This new protection strategy allowed for the upscaling of the synthesis of 1-propyl-8-p-sulfophenylxanthine (PSB-1115), a selective A(2B) antagonist. PMID- 14761206 TI - Inhibition of adenosine deaminase by novel 5:7 fused heterocycles containing the imidazo[4,5-e][1,2,4]triazepine ring system: a structure-activity relationship study. AB - As part of a program to explore structure-activity relationships for the extremely tight binding inhibition characteristics of coformycins to adenosine deaminase, a series of analogues (1a-1h) containing the imidazo[4,5 e][1,2,4]triazepine ring system has been synthesized and screened in vitro against a mammalian adenosine deaminase for inhibitory activity. While compounds 1a and 1b were synthesized in five steps starting from 4-nitroimidazole, others were derived from 1a through simple exchange reactions with the appropriate alcohols. The observed kinetics profiles and K(i) values suggest that the target compounds are competitive inhibitors that bind 6-9 orders of magnitude less tightly to the enzyme. Compounds 1c and 1d were the most active in the series with K(i)'s ranging from 12 to 15 microM. PMID- 14761207 TI - Macrocyclic polyamines deplete cellular ATP levels and inhibit cell growth in human prostate cancer cells. AB - In solid tumors, when O(2) partial pressure drops below 10 mmHg, ATP levels rapidly decrease due to the Warburg effect. It is known that certain macrocyclic polyamines catalyze the chemical hydrolysis of ATP with release of inorganic phosphate. Since tumor cells have diminished ATP levels as compared to normal cells, we attempted to deplete cellular ATP with macrocyclic polyamines in an effort to inhibit tumor cell proliferation. Five macrocyclic polyamines, related to the budmunchamine family of alkaloids, were prepared by total synthesis. They were the [17]-N(4) macrocycle 1, the [16]-N(4) macrocycle 20, the [18]-N(4) macrocycle 13, the [20]-N(5) macrocycle 8, and the [13]-N(3) macrocycle 17. Each one of them hydrolyzed ATP in vitro with release of P(i); the largest ring macrocycle 8 was the most efficient catalyst, while the smallest ring macrocycle 17 was the least efficient (P(i) released in these runs was on the order of 40 100 microM). The linear polyamine spermine had no hydrolytic effect on ATP. The macrocycles were found to be cytotoxic when assessed by means of a MTT assay against two human prostate cell lines, DuPro and PC-3, with resultant ID(50) values ranging between 0.5 and 1.8 microM. Colony forming efficiency (CFE) assays performed on DuPro cells, where the macrocycles were used in a concentration range of 1-8 microM, confirmed the cytotoxic effect of each macrocycle. Each killed 3-4 log of DuPro cells. The smallest ring 17 was the least cytotoxic after 24 h of incubation, although after 144 h of incubation it showed significant cytotoxicity at 8 microM. The macrocycles were equally efficient in depleting the intracellular ATP pools; after a 24 h incubation with each macrocycle other than 17 at 1-8 microM concentrations, cellular ATP concentrations were decreased by 3 orders of magnitude. The decrease in ATP levels was more pronounced after a 72 h incubation, when even 17 reduced ATP by 2 orders of magnitude. A linear pentamine of established cytotoxicity was without effect on the ATP pools. The macrocycles depleted almost entirely the intracellular pools of polyamines and were efficiently taken up by cells. A rough correlation could be established between the cytotoxic effect of the macrocyclic polyamines and their ATP-ase like activity in the DuPro cell line. As ATP is a scarce metabolite in cancer cells, where it can only be replenished through the very ATP-inefficient glycolytic pathway; macrocyclic polyamines appear to be promising new anticancer agents. PMID- 14761208 TI - Phe-Gly dipeptidomimetics designed for the di-/tripeptide transporters PEPT1 and PEPT2: synthesis and biological investigations. AB - A series of five Phe-Gly dipeptidomimetics containing different amide bond replacements have been synthesized in a facile way from the readily available unsaturated ketoester 1, and their affinities for the di-/tripeptide transporters hPEPT1 (Caco-2 cells) and rPEPT2 (SKPT cells) were tested. The compounds contained the amide bond isosteres ketomethylene (2a), (R)- and (S) hydroxyethylidene (3a and 4a), and (R)- and (S)-hydroxyethylene (5a and 6a) to provide information on the conformational and stereochemical requirements for hPEPT1 and rPEPT2 affinity. The affinity studies showed that for rPEPT2 there is no significant difference in affinity between the ketomethylene isostere 2a and the natural substrate Phe-Gly (K(i) values of 18.8 and 14.6 microM, respectively). Also the affinities for hPEPT1 are in the same range (K(i) values of 0.40 and 0.20 mM, respectively). This corroborates earlier findings that the amide bond as such is not essential for binding to PEPTX, but the results also reveal possible differences in the binding of ketomethylene isosteres to hPEPT1 and rPEPT2. The trans-hydroxyethylidene and hydroxyethylene isosteres proved to be poor substrates for PEPTX. These results provide new information about the importance of flexibility and of the stereochemistry at the C(4)-position for this class of compounds. Furthermore, the intracellular uptake of 2a-4a in Caco-2 cells was investigated, showing a 3-fold reduction of the uptake of 2a in the presence of the competetive inhibitor Gly-Pro, indicating contribution from an active transport component. No active uptake of 3a and 4a was observed. Transepithelial transport studies also indicated active transport of 2a across Caco-2 monolayers. PMID- 14761209 TI - Importance of phenolic address groups in opioid kappa receptor selective antagonists. AB - In vitro characterization and comparison of JDTic, its dehydroxy analogue and nor BNI, and its dehydroxy analogue demonstrates that the N-substituted 3,4-dimethyl (3-hydroxyphenyl)piperidine-derived antagonist, JDTic, relies more heavily on its phenol address group for affinity and antagonist activity relative to the corresponding naltrexone derived antagonists, nor-BNI. The structural flexibility of the former class of compound relative to the latter is postulated to underlie the difference. PMID- 14761237 TI - Developing countries do not have the same needs as developed countries for AIDS control: interest of future vaccines. PMID- 14761239 TI - Therapeutic vaccines for substance dependence. AB - Immunotherapies are under development as a new approach to the treatment of substance dependence. The drugs of abuse currently being tested using this new approach are nicotine, cocaine, phencyclidine and methamphetamine. In laboratory animal models, a range of immunotherapies, including vaccines, monoclonal antibodies and catalytic antibodies, have been shown to reduce drug seeking. In human clinical trials, cocaine and nicotine vaccines have been shown to induce antibody titers while producing few side effects. Studies in humans determining how these vaccines interact in combination with their target drug are underway. Overall, immunotherapy offers a range of potential treatment options: drug treatment, as well as the treatment of overdose, prevention of brain or cardiac toxicity and fetal protection in pregnant drug abusers. PMID- 14761240 TI - MMR vaccine and autism: an update of the scientific evidence. AB - An hypothesis published in 1998 suggested that measles-mumps-rubella vaccine may cause autism as a result of persistent measles virus infection of the gastrointestinal tract. Results of early studies were not supportive and in 2001 a review by the Institute of Medicine concluded that the evidence favors the rejection of a causal relationship at the population level between measles-mumps rubella vaccine and autistic spectrum disorder. Studies published since the Institute of Medicine report have continued not to find an increased risk of autistic spectrum disorder associated with measles-mumps-rubella. The vaccine also has not been found to be associated with a unique syndrome of developmental regression and gastrointestinal disorders. The evidence now is convincing that the measles-mumps-rubella vaccine does not cause autism or any particular subtypes of autistic spectrum disorder. PMID- 14761241 TI - Antibody regulation of Tcell immunity: implications for vaccine strategies against intracellular pathogens. AB - Intracellular microbial pathogens cause a plethora of diseases that pose a huge public health challenge. Efficacious prophylactic vaccines are needed to protect the population from this myriad of infectious diseases. Contemporary approaches to vaccine design are guided by the immunobiological paradigm that extracellular pathogens are controlled principally by humoral immunity, involving specific antibodies, whereas host protection against intracellular pathogens requires effectors of cell-mediated immunity. However, this distinct T-helper (Th) type 1 and 2 paradigm of host defense has encountered a major challenge due to the reality that most antigens or vaccines induce mixed immune responses comprising of both humoral and CMI effectors. Besides, the true functional independence of antibodies and T-cells under in vivo physiologic conditions is uncertain. Recent findings have revealed that antibodies exert a significant immunoregulatory effect on T-cell immunity. Thus, a robust and protective T-cell memory response against microbial pathogens such as Chlamydia and Mycobacteria require an effective primary humoral immune response characterized by specific antibody isotypes whose role is to modulate Th1 activation via Fc receptors (FcR) by facilitating a rapid uptake, processing and presentation of pathogen-derived antigens for an enhanced T-cell response. These findings have crystallized into a paradigm shift in host defense wherein different components of the apparently disparate mixed immune responses elicited against a microbial pathogen function concertedly to maximize the principal effector mechanism. This review focuses on the essential role of both arms of the immune system in controlling intracellular microbial pathogens, especially the regulatory role of FcR-mediated antibody function in optimizing the induction of a protective Th1 response. The immunobiological implications are discussed in the context of vaccine design, delivery and evaluation against intracellular microbial pathogens of bacteria, fungi and parasitic origin. PMID- 14761242 TI - The threat and prospects for control of an influenza pandemic. AB - Influenza constitutes the most widespread and significant respiratory infectious disease in the world, resulting in increased morbidity, mortality and economic loss each epidemic year. Pandemic influenza is a worldwide epidemic usually caused by a new virus variant to which the majority of the population has no immunity. As demonstrated in the devastating pandemic of 1918 to 1919, a pandemic virus may infect 30 to 50% of the worlds population and kill 1 to 2% of those infected. Pandemic control must be a concerted and co-ordinated world strategy and under the auspices of the World Health Organization, pandemic preparedness plans have been formulated, including: intensified surveillance for more rapid identification of new reassortant viruses with potential human virulence and infectivity, laboratory characterization of the new viruses so that vaccine may be prepared, development of techniques for more rapid vaccine production and the manufacture and stock piling of antiviral drugs. The H5N1 outbreak of virulent chicken influenza in 1997 in Hong Kong which resulted in the deaths of six of 18 infected persons serves as a wake-up call. Should such a virus attain high transmissibility in humans, a pandemic of tragic proportions might ensue. Even though the timing of onset of the next pandemic cannot be precisely predicted, world governments must understand the urgency of the problem and increase funding for influenza pandemic control. PMID- 14761243 TI - Lipid core peptide technology and group A streptococcal vaccine delivery. AB - The antiphagocytic surface M protein of group A streptococcus has been widely studied as the major candidate antigen for a vaccine to prevent group A streptococcus infection. Approaches that have proven to be effective in animal models include the use of multi-epitope vaccines incorporating highly variable amino terminal serotypic determinants, those based on the carboxy terminal conserved region and combination vaccines incorporating both serotypic and conserved region determinants of the M protein. The use of lipid core peptide technology is at the forefront of this research in the quest to develop a broad strain protective vaccine that can be delivered via the mucosal route, stimulating mucosal and systemic immunity. This review aims to cover the various strategies and technologies that have been investigated with regard to group A streptococcus vaccine design and development. PMID- 14761244 TI - Genome-derived vaccines. AB - Vaccine research entered a new era when the complete genome of a pathogenic bacterium was published in 1995. Since then, more than 97 bacterial pathogens have been sequenced and at least 110 additional projects are now in progress. Genome sequencing has also dramatically accelerated: high-throughput facilities can draft the sequence of an entire microbe (two to four megabases) in 1 to 2 days. Vaccine developers are using microarrays, immunoinformatics, proteomics and high-throughput immunology assays to reduce the truly unmanageable volume of information available in genome databases to a manageable size. Vaccines composed by novel antigens discovered from genome mining are already in clinical trials. Within 5 years we can expect to see a novel class of vaccines composed by genome predicted, assembled and engineered T- and Bcell epitopes. This article addresses the convergence of three forces--microbial genome sequencing, computational immunology and new vaccine technologies--that are shifting genome mining for vaccines onto the forefront of immunology research. PMID- 14761245 TI - Serologic correlates of protection for evaluating the response to meningococcal vaccines. AB - Meningococci cause serious disease worldwide and the organism remains the most common cause of bacterial meningitis in children and young adults. The only effective means of controlling disease is through vaccination. Although polysaccharide vaccines have been available for serogroup A, C, Y and W135 for many years, serogroup C polysaccharide-protein conjugate vaccines have only recently been licensed in many countries. Conjugate vaccines for combinations of serogroup A, C, Y and W135 are progressing through clinical trials and major efforts are being made to develop a safe and efficacious vaccine against serogroup B. To assess the quality of the immune response after vaccination, laboratory correlates of protection are needed. For serogroups A and C, serum bactericidal antibody is a well established predictor for protection but for serogroup B, other mechanisms besides serum bactericidal antibody may also be involved in conferring protection against disease. The serologic correlates of protection for evaluating the response to meningococcal vaccines are described in this review. PMID- 14761246 TI - Toxicity and potency evaluation of pertussis vaccines. AB - Current methods for determining the potency and toxicity of pertussis vaccines are outdated and require improvement. The intracerebral challenge test is effective for determining the potency of whole-cell vaccines but is objectionable on animal welfare and technical grounds. The same applies to its modification for assaying acellular pertussis vaccines. Respiratory challenge methods offer an interim solution pending establishment of validated in vitro correlates of protection, for example nitric oxide induction. Their evaluation is being promoted by the World Health Organization through the Pertussis Vaccines Working Group. Current toxicity assays based on weight gain and histamine sensitization of mice are imprecise and need replacement. Limits need to be established for specific toxin content of both acellular and whole-cell vaccines and should be supported by specific assays. More precise methods based on determination of ribosyltransferase activity in tandem with receptor-binding assays are under evaluation. Genome sequence data and the use of gene microarrays to screen responses triggered by vaccine components may also provide leads to improved methods for assessing both toxicity and immunogenicity. PMID- 14761247 TI - Increased arterial stiffening and thickening in the paretic lower limb in patients with hemiparesis. AB - Atherosclerosis has two key components, thickening and stiffening of arterial wall. These parameters are quantified ultrasonographically by IMT (intima-media thickness) and PWV (pulse wave velocity). In the present study, we determined the FA IMT (IMT of the bilateral femoral artery) and PWV of femoral-ankle (PWV fa) and brachial-ankle (PWV ba) segments in order to examine whether the degree of atherosclerosis is different between paretic and non-paretic lower limbs in 24 patients with hemiparesis. The values of PWV fa, PWV ba and FA IMT were all significantly greater on the paretic than the non-paretic side. Furthermore, significant decreases in masses of muscle, bone and fat, determined by dual energy X-ray absorptiometry, were observed in paretic lower limbs compared with the non-paretic side. PWV fa correlated significantly and negatively with muscle mass ( r =-0.488, P =0.0004) and tended to correlate negatively with BMC (bone mineral content; r =-0.264, P =0.069) when statistical analyses were performed with the paretic and non-paretic sides together. Multiple regression analysis elucidated that the muscle mass was associated significantly with PWV fa and PWV ba, independent of age, duration after cerebrovascular accident, gender, bone and fat mass and FA IMT. The muscle mass was still associated with increased PWV fa and PWV ba when multivariate analysis was conducted independently in the paretic and non-paretic sides. In summary, our results indicated that arterial thickening and stiffening were greater on the paretic than the non-paretic side and suggested that a decrease of muscle mass might be associated with increased arterial stiffening in the paretic lower limb. PMID- 14761248 TI - An efficient isocratic separation of hydroxycinnamates and their corresponding benzoates from microbial and plant sources by HPLC. AB - A rapid HPLC-based separation method was developed to analyse phenolic flavour components. In the present study, ferulic acid, 4-coumaric acid, 4-hydroxybenzoic acid, 4-hydroxybenzaldehyde, vanillic acid and vanillin were chromatographed on various C(18) columns (Prodigy ODS2, Synergi Hydro-RP, Lichrosorb and Columbus). A dual-wavelength UV detector was used for the precise identification of the eluted components. An isocratic elution with aqueous trifluoroacetic acid (1 mM)/methanol (17:8) at a flow rate of 1.0 ml/min separated all the above six phenolic compounds within 21 min on a C(18) reverse-phase column (Synergi Hydro RP) with stable baseline resolution. PMID- 14761250 TI - Regional distribution of fatiguing illnesses in the United States: a pilot study. AB - BACKGROUND: Chronic fatigue syndrome (CFS) is a debilitating illness with no known cause or effective therapy. Population-based epidemiologic data on CFS prevalence are critical to put CFS in a realistic context for public health officials and others responsible for allocating resources. METHODS: We conducted a pilot random-digit-dialing survey to estimate the prevalence of fatiguing illnesses in different geographic regions and in urban and rural populations of the United States. This report focuses on 884 of 7,317 respondents 18 to 69 years old. Fatigued (440) and randomly selected non-fatigued (444) respondents completed telephone questionnaires concerning fatigue, other symptoms, and medical history. RESULTS: We estimated 12,186 per 100,000 persons 18 to 69 years of age suffered from fatigue lasting for at least 6 months (chronic fatigue), and 1,197 per 100,000 described an illness that, though lacking clinical evaluation, met criteria for CFS (CFS-like). Chronic fatigue and CFS-like illness were more common in rural than in urban populations, although the differences were not significant. The prevalence of these fatiguing illnesses did not differ meaningfully among the four regions surveyed, and no significant geographic trends were observed. CONCLUSIONS: This investigation estimated that nearly 2.2 million American adults suffer from CFS-like illness. The study also suggested the need to focus future investigations of fatigue on populations with lower incomes and less education. There was no evidence for regional differences in the occurrence of fatiguing illnesses. PMID- 14761251 TI - A biocompatible magnetic film: synthesis and characterization. AB - BACKGROUND: Biotechnology applications of magnetic gels include biosensors, targeted drug delivery, artificial muscles and magnetic buckles. These gels are produced by incorporating magnetic materials in the polymer composites. METHODS: A biocompatible magnetic gel film has been synthesized using polyvinyl alcohol. The magnetic gel was dried to generate a biocompatible magnetic film. Nanosized iron oxide particles (gamma-Fe2O3, ~7 nm) have been used to produce the magnetic gel. RESULTS: The surface morphology and magnetic properties of the gel films were studied. The iron oxide particles are superparamagnetic and the gel film also showed superparamagnetic behavior. CONCLUSION: Magnetic gel made out of crosslinked magnetic nanoparticles in the polymer network was found to be stable and possess the magnetic properties of the nanoparticles. PMID- 14761252 TI - Clinical and Molecular Allergy: a new open access journal that addresses rapidly evolving information in the field of allergy and immunology. PMID- 14761253 TI - Vasa vasorum in plaque angiogenesis, metabolic syndrome, type 2 diabetes mellitus, and atheroscleropathy: a malignant transformation. AB - BACKGROUND: Vascularization is an exciting and complex mechanism involving angiogenesis and arteriogenesis. The metabolic syndrome (MS) and type 2 diabetes mellitus (T2DM) are associated with multiple metabolic toxicities, which result in reactive oxygen species (ROS) due to an elevated tension of oxidative-redox stress and an accelerated atherosclerosis termed atheroscleropathy. RESULTS: This atheroscleropathy is associated with accelerated angiogenesis within the vulnerable, thin-cap fibro-atheroma, prone to rupture resulting in acute coronary syndromes (ACS). The resulting intimopathy with its neovascularization due to angiogenesis of the adventitial vasa vasorum (Vv) is prone to intraplaque hemorrhage (IPH). These IPH are associated with destabilization of the vulnerable plaques resulting in plaque erosion and plaque rupture resulting in ACS. In atheroscleropathy the adventitial Vv invades the plaque in a malignant-like fashion and concurrently is associated with chronic inflammation, as macrophages are being deposited within the shoulder regions of these vulnerable plaques. These angiogenic Vv provide a custom delivery vascular network for multiple detrimental substrates, which further accelerates the growth of these vulnerable plaques and atheroscleropathy. There exists a vascularization paradox in MS and T2DM, in that, angiogenesis within the plaque is induced and arteriogenesis is impaired. CONCLUSION: This review will attempt to provide a database of knowledge regarding the vascularization process (angiogenesis and arteriogenesis) and its mechanisms to better understand the increased cardiovascular risk and the increased morbidity and mortality associated with MS and T2DM. PMID- 14761255 TI - Simulating market dynamics: interactions between consumer psychology and social networks. AB - Markets can show different types of dynamics, from quiet markets dominated by one or a few products, to markets with continual penetration of new and reintroduced products. In a previous article we explored the dynamics of markets from a psychological perspective using a multi-agent simulation model. The main results indicated that the behavioral rules dominating the artificial consumer's decision making determine the resulting market dynamics, such as fashions, lock-in, and unstable renewal. Results also show the importance of psychological variables like social networks, preferences, and the need for identity to explain the dynamics of markets. In this article we extend this work in two directions. First, we will focus on a more systematic investigation of the effects of different network structures. The previous article was based on Watts and Strogatz's approach, which describes the small-world and clustering characteristics in networks. More recent research demonstrated that many large networks display a scale-free power-law distribution for node connectivity. In terms of market dynamics this may imply that a small proportion of consumers may have an exceptional influence on the consumptive behavior of others (hubs, or early adapters). We show that market dynamics is a self-organized property depending on the interaction between the agents' decision-making process (heuristics), the product characteristics (degree of satisfaction of unit of consumption, visibility), and the structure of interactions between agents (size of network and hubs in a social network). PMID- 14761256 TI - World-size global markets lead to economic instability. AB - Economic and cultural globalization is one of the most important processes humankind has been undergoing lately. This process is assumed to be leading the world into a wealthy society with a better life. However, the current trend of globalization is not unprecedented in human history, and has had some severe consequences in the past. By applying a quantitative analysis through a microscopic representation we show that globalization, besides being unfair (with respect to wealth distribution), is also unstable and potentially dangerous as one event may lead to a collapse of the system. It is proposed that the optimal solution in controlling the unwanted aspects and enhancing the advantageous ones lies in limiting competition to large subregions, rather than making it worldwide. PMID- 14761257 TI - Iterated learning: a framework for the emergence of language. AB - Language is culturally transmitted. Iterated learning, the process by which the output of one individual's learning becomes the input to other individuals' learning, provides a framework for investigating the cultural evolution of linguistic structure. We present two models, based upon the iterated learning framework, which show that the poverty of the stimulus available to language learners leads to the emergence of linguistic structure. Compositionality is language's adaptation to stimulus poverty. PMID- 14761258 TI - Evolution of an optimal lexicon under constraints from embodiment. AB - Research in language evolution is concerned with the question of how complex linguistic structures can emerge from the interactions between many communicating individuals. Thus it complements psycholinguistics, which investigates the processes involved in individual adult language processing, and child language development studies, which investigate how children learn a given (fixed) language. We focus on the framework of language games and argue that they offer a fresh and formal perspective on many current debates in cognitive science, including those on the synchronic-versus-diachronic perspective on language, the embodiment and situatedness of language and cognition, and the self-organization of linguistic patterns. We present a measure for the quality of a lexicon in a population, and derive four characteristics of the optimal lexicon: specificity, coherence, distinctiveness, and regularity. We present a model of lexical dynamics that shows the spontaneous emergence of these characteristics in a distributed population of individuals that incorporate embodiment constraints. Finally, we discuss how research in cognitive science could contribute to improving existing language game models. PMID- 14761259 TI - Emergent patterns of mate choice in human populations. AB - We present a model of human mate choice that shows how realistic population-level patterns of assortative mating can self-organize and emerge from the behavior of individuals using simple mate search rules. In particular, we model plausible psychological mechanisms for mate search and choice in a realistic social ecology. Through individual interactions, patterns emerge that match those observed in typical human societies, particularly with regard to correlated quality levels within couples, distributions of the ages at which couples mate, and effects of skewed sex ratios on these mating age distributions. PMID- 14761260 TI - Emergent properties in small-scale societies. AB - A multi-agent simulation is used to explore the relationship between the micro and the macro levels in small-scale societies. The simulation demonstrates, using an African hunter-gatherer group (the !Kung san) as a case study, the way in which population stability may arise from culturally framed, micro-level decision making by women about spacing of births. According to the simulation, population stability as an emergent property has different implications, depending on resource density. Data on Australian hunter-gatherer groups are presented that support the implications of the simulation. !Kung san micro-level cultural rules on incestuous marriages are shown to have macro-level consequences in the form of marriages between residential camps. Between-camp marriages have significant implications for access to resources and thereby for population dynamics of the group as a whole. PMID- 14761261 TI - The evolution of social behavior in the prehistoric American southwest. AB - Long House Valley, located in the Black Mesa area of northeastern Arizona (USA), was inhabited by the Kayenta Anasazi from circa 1800 B.C. to circa A.D. 1300. These people were prehistoric precursors of the modern Pueblo cultures of the Colorado Plateau. A rich paleoenvironmental record, based on alluvial geomorphology, palynology, and dendroclimatology, permits the accurate quantitative reconstruction of annual fluctuations in potential agricultural production (kg maize/hectare). The archaeological record of Anasazi farming groups from A.D. 200 to 1300 provides information on a millennium of sociocultural stasis, variability, change, and adaptation. We report on a multi agent computational model of this society that closely reproduces the main features of its actual history, including population ebb and flow, changing spatial settlement patterns, and eventual rapid decline. The agents in the model are monoagriculturalists, who decide both where to situate their fields and where to locate their settlements. PMID- 14761270 TI - [Studies on etiology and vaccines of hepatitis E]. PMID- 14761271 TI - [Significance of serological markers and virological marker for hepatitis E in rhesus monkey model]. AB - OBJECTIVE: To evaluate the serological markers and biological marker in the diagnosis of hepatitis E infection in a rhesus monkey model. METHODS: 86 rhesus monkeys had been infected with different doses of HEV. Hence, they were taken sequential blood samples at intervals up to 86 weeks for 4 hepatitis E virus (HEV) specific antibody assays (E2-IgM, E2-IgG, GL-IgG, and YES-IgG), and nucleic acid assay. RESULTS: All the animals produced E2-IgG and all but one also produced E2-IgM and excreted the virus in stool, whereas positive rate of GL-IgG and YES IgG were low and correlated with virus level. Hepatitis occurred over a period of 4 weeks (between 3 an 7 weeks) after infection. Virological marker occurred mainly during incubation period and declined rapidly after onset of hepatitis. Seroconversion of E2-IgM occurred before onset of hepatitis in 70% monkeys and declined rapidly up to 50% of peak value after 4 weeks. E2-IgM seroconversion was closely paralleled by E2-IgG; however, E2-IgG persisted in all animals for the entire duration of experiment of up to 86 weeks. Production of GL IgG and YES-IgG was delayed by one week after the E2 antibodies, these antibodies showed a transient occurrence and seroprevalence declined to 50% of the peak value over a period of 12 weeks. CONCLUSION: E2-IgM might be used as a suitable acute hepatitis E marker, and E2-IgG as a suitable epidemiological marker. The seroconversion or titer elevation of GL-IgG and YES-IgG antibodies probably used to confirm the infection. The viral markers are optional for early diagnosis. PMID- 14761272 TI - [Investigation of sub-clinical infection of hepatitis E virus in blood donors]. AB - OBJECTIVE: To understand the prevalence of sub-clinical infection of hepatitis E virus (HEV) among blood donors. METHODS: A cluster sampling strategy was used to sample all blood donors from July to August in 2002 in Beijing. Their blood was tested for IgM and IgG antibody against HEV. RESULTS: The prevalence of anti-HEV IgM among blood donors in Beijing was 1.74%. The rate of abnormal alanine aminotransferase (ALT) in anti-HEV IgM positive donors is significantly higher than anti-HEV IgM negative donors. Among all ALT abnormal donors, 2.68% can be associated with HEV sub-clinical infection. The percentage is similar with HBV but higher than HCV. CONCLUSION: There are sub-clinical infection of HEV among blood donors, which is one of the cause of abnormal ALT in the donors. PMID- 14761273 TI - [Detection of hepatitis E virus on a blood donor and its infectivity to rhesus monkey]. AB - OBJECTIVE: To understand the infectivity and pathogenicity of the plasma of hepatitis E virus (HEV) viremia to primate animals. METHODS: RNA fragment of HEV genotype IV was detected on one healthy donor who was positive for anti-HEV IgM and negative for anti-HEV IgG. Then 10 ml plasma from above donor was transfused to rhesus monkey to observe its infectivity and pathogenicity. RESULTS: Acute hepatitis E was developed in rhesus monkey who accept HEV RNA positive plasma. It was confirmed by virological, immunological, biochemical and histopathological data. CONCLUSION: Acute hepatitis E can be induced by plasma transfusion of HEV viremia, which indicate the possibility of transfusion transmitted hepatitis E PMID- 14761274 TI - [Side-effects and management of continuous MARS artifical liver for patients]. PMID- 14761275 TI - [Evaluation of the reliability of three different hepatitis E diagnosis reagents on the clinical acute hepatitis E]. AB - OBJECTIVE: To evaluate the reliability of different hepatitis E diagnosis reagent tests on the acute hepatitis E. METHODS: Three acute hepatitis E diagnosis tests, E2-IgM (Wantai, China), GL-IgM and GL-IgG (Genelabs, Singapore) were compared for their reliability in a sera panel composed by 273 healthy individuals and 525 hepatitis. RESULTS: The specificity of E2-IgM on the diagnosis of acute hepatitis E was 100.0%, it was significantly higher than GL-IgM (96.7%) and GL-IgG (85.4%). The sensitivity of E2-IgM and GL-IgG were 97.9% and 93.8% respectively, both significantly higher than GL-IgM (72.9%). Among 65 acute hepatitis cases being positive on GL-IgM test but negative on E2-IgM, 58 (89.2%) cases were found to be positive with anti-hepatitis A virus IgM, it indicated that the GL-IgM test might be interfered by other IgM antibodies on serum. CONCLUSION: E2-IgM is a good test for the diagnosis of acute hepatitis E. PMID- 14761276 TI - [An investigation on the transmission routes and early diagnosis of intrauterine infection induced by hepatitis B virus]. AB - OBJECTIVES: To analyze the relationship between the fetus infection and HBV M, HBV DNA in amniotic fluid, umbilical cord blood, maternal blood and placenta, and to explore the mechanism of vertical transmission of HBV. METHODS: Immunonetric assay and nucleic acid amplification hybri-comb were used. Both HBV M and HBV DNA were detected in amniotic fluid, vein blood, umbilical cord blood for each of 65 HBV-positive women in their different gestational periods, while immunohistochemical analysis was carried out on the tissue of placenta, liver, lung or heart from each abortive fetus/dead infant in the case. RESULTS: For all of the 65 HBsAg-positive women in their different gestational periods, the detected positive rate of HBsAg was 21.50% in amniotic fluid, and 20.00% in umbilical blood. The positive rate of HBsAg, HBeAg, Anti-HBc and HBV DNA detected in blood, amniotic fluid and umbilical blood was 6.15%. The cases with positive HBsAg, Anti-HBe, Anti-HBc and negative HBV DNA were in a percentage of 13.85%. Immunohistochemical analysis on placentas after birth/abortion as well as the tissues of livers, lungs, hearts of the fetuses/dead infants in 4 cases of pregnant women with positive HBsAg, HBeAg, Anti-HBc or HBV DNA in blood, amniotic fluid or umbilical blood showed that HBsAg, HBcAg positive cells in the scope could be seen in every layer of the tissue of placenta, in the hepatic/pulmonary tissue, but not in the cardiac tissue. CONCLUSION: The infection in amniotic fluid or placenta relates to HBV infection in fetus; intrauterine HBV may result in infection in organs such as blood, liver, or lung of a fetus; infection in the amniotic fluid may be another key route of the intrauterine infection of fetus, and the detection on HBV M or HBV DNA in amniotic may be used as one of diagnostic proofs of HBV infection of fetus in its early stage. PMID- 14761277 TI - [Susceptibility change to HBV in primary culture of first trimester human fetal hepatocytes]. AB - OBJECTIVES: By culturing primary early (8 to 12-weeks-old) human fetal hepatocytes with different conditions, to study the status of cell susceptibility to HBV. METHODS: During primary culture of 10-weeks-old human fetal hepatocytes with serum-free medium adding different differentiation-induced ingredients, to inoculate cell with HBV at certain time. Cell shape, function and markers of HBV infection are measured. RESULTS: 6 days after seeding, markers of mature hepatocytes are observed in cells cultured with 2.5mmol/L phenobarbital sodium, and these cells show susceptibility to HBV. Other ingredients cannot render hepatocytes susceptible to HBV. CONCLUSION: Phenobarbital sodium induces differentiation and susceptibility to HBV in primary culture of early human fetal hepatocytes. PMID- 14761278 TI - [Study on the replication of Hepatitis B Virus in primary hepatocytes from heterogenous species]. AB - OBJECTIVES: By studying the possibility and degree of the replication and expression of human hepatitis B virus (HBV) genes in normal liver cells from heterogenous species, such as primary duck hepatocytes (PDH) and primary rat hepatocytes (PRH), to investigate the species-specificity of HBV infection and replication. METHODS: PDH and PRH isolated by in situ perfusion with low concentration collagenase were transfected with complete HBV genome by electroporation (transfection group, about 1.19 10(12) copies of linear HBV DNA/1 10(7) PRH/PDH). From 1 day to 15 days after transfection, HBsAg and HBeAg in the supernatants and lysates of PRH/PDH were measured with IMX System, HBcAg was assayed with western blotting, Immunol dot blotting and Immunocytochemistry. Moreover, HBV S-mRNA and X-mRNA were tested with RT-PCR. Meanwhile, replicative intermediates of HBV DNA were analyzed by Southern blotting and Dot blotting. PRH/PDH electroporated only was used as control group. RESULTS: HBsAg in the lysates of transfected PDH group were 15.24 (1day), 14.55 (3 days) and 5.13 ( 5 days; P/N values, positive>or=2.1), HBeAg all was negative (<2.1), and both were negative in the supernatants of transfected group. Viral antigen production in transfected rat hepatocytes: HBsAg in the lysates of transfected hepatocytes was positive, P/N values ranging from 2.17 to 93.41, The average P/N values was 14.74+/-31.82, and could be maintained for 15 days after transfection. Whereas, HBsAg in the supernatants of transfected group was only found positive on 1 day after transfection, which P/N value was 6.66. HBeAg and HBcAg in the lysates of PRH of transfection group were positive during the first 3 days following transfection, P/N values was around 2.8. The total amount of HBV DNA in the transfected PDH and PRH groups was strongly positive by dot blotting, whereas that of the control group was negative. Southern blot analysis of intracellular total HBV DNA indicated that there were relaxed circular (RC), covalently closed circular (ccc) and single-stranded (SS) HBV DNA replicative intermediates in the transfected PDH and PRH groups, there was no integrated HBV DNA in the cellular genome. Control groups were negative at all. CONCLUSION: Expression of HBV genes and production can occur in hepatocytes from nonmammalian species or mammalian species, which strongly supports the idea that replication of HBV has no critical species-specificity, and yet it depends on the endoenvironment of hepatocyte. PMID- 14761279 TI - [A study on the mutation of P gene region including YMDD motif in hepatitis B virus]. AB - OBJECTIVE: To investigate the primary structure and heterogenenity of P gene region including YMDD motif in hepatitis B virus. METHODS: From serum samples collected from 4 patients who had never been treated with anti-viral drugs, DNA fragments of 1057bp long of P gene were amplified and cloned into pUC19. Twenty positive clones were chosen randomly from each sample. The YMDD motif mutation was detected by mismatched PCR-RFLP. Finally last ten positive clones of two samples were sequenced. RESULTS: Nucleotide mutation rates among clones of Sample 1 and 2 were 0.3% - 1.1%, 0.4% - 1.7%, respectively. Among 80 clones, the variations from YMDD to YMGD were revealed in two clones. CONCLUSION: There are HBV quasispecies in the P gene region including YMDD motif of hepatitis B virus and a novel mutation of YMDD motif in the sera of patients without being therapied by anti-viral drugs. PMID- 14761280 TI - [Perindopril attenuates the progression of CCl4-inducing rat hepatic fibrosis]. AB - OBJECTIVES: The aim of the present study was to determine the effects of angiotensin-converting enzyme inhibitor, perindopril, on the progression of rat hepatic fibrosis induced by CCl4. METHODS: Male wistar rats weighting about 250g were treated with perindopril (2mg/kg, daily gavage), except for model group and control group. After 4, 6 weeks, morphological examination was based on microscopy. RT-PCR was utilized to detect gene expression of angiotensin II type 1 receptor (AT1 receptor) in the liver. Meanwhile, the protein expressions of AT1 receptor, transforming growth factor beta 1 (TGF-beta1) and platelet-derived growth factor-BB (PDGF-BB) in liver tissue were examined by Western blot. The activity of matrix metalloproteinase-2 (MMP-2) was assessed by zymography. Serum laminin (LN) and hyaluronic acid (HA) were measured using radio-immunity technique. RESULTS: RT-PCR and Western blot revealed that there was a up regulation in AT1 receptor expression in model group compared with control group. Perindopril treatment significantly reduced mean fibrosis score, messenger RNA and protein levels of AT1 receptor, protein levels of TGF-beta1 and PDGF-BB, Serum levels of HA and LN, and MMP-2 activity. CONCLUSION: These results suggest that angiotensin II may play an important role in fibrosis of liver. Perindopril may have a inhibiting effect on CCl4-induced hepatic fibrogenesis of rat. PMID- 14761281 TI - [Inhibitory effects of antisense oligonucleotides on VEGF gene expression by human hepatocellular carcinoma cells]. AB - OBJECTIVE: To investigate the inhibitory effects of antisense oligonucleotides to different sequences on VEGF gene expression by human hepatoma cells. METHODS: SMMC7721 cells were cultured under normoxic or hypoxic conditions for 24 h, followed by being transfected with different antisense oligonucleotides (A06513 to cap structure, A06514 to translation initiation, A06515 to Exon-3 and A06516 to translation terminal). The total RNAs from the cells were extracted and the VEGF expression were examined with RT-PCR. The relative concentrations of VEGF transcripts in SMMC772 cells from different groups were determined using GAPDH (glyceraldehyde-3-phosphate dehydrogenase) cDNA as internal standard. RESULTS: In response to the hypoxic challenge, SMMC7721 cells upregulated VEGF mRNA; Comparative to the control (no oligonucleotides), A06513, A06514, A06515, and A06516 had obvious sequence-specific inhibitory effect on VEGF gene expression, with the ratio of VEGF over GAPDH of 0.49+/-0.08, 0.71+/-0.12, 0.72+/-0.11 and 0.86+/-0.12, respectively (F=12.21, P< 0.05). A06513 showed the strongest inhibitory effect (P<0.01). CONCLUSION: The antisense oligonucleotides complementary to VEGF cap structure, may become a potential alternative for antisense gene therapy of HCC. PMID- 14761282 TI - [Transitional CK19 positive cells-a new possible marker of hepatic precancerous lesion]. AB - OBJECTIVE: To investigate the relationship between Transitional CK19 positive cells and hepatic precancerous lesion in chronic hepatitis B patients. METHODS: We observed the expression of CK19 in liver tissue of chronic hepatitis B patients by LSAB immunohistochemical staining, and examined serum AFP and ultrasonography one time per 3 months for one year. RESULTS: We observed a population of CK19 positive cells-with size and structure between those of human oval cells and mature hepatocytes-that usually occurred along with oval-cell proliferation. It was suggested that these transitional cells may partly account for the elevation of serum AFP. One patient occurred hepatic carcinoma, another patient had low-echogenic nodules in liver parenchyma within the 1 year follow-up period. CONCLUSION: Transitional CK19 positive cells could be regarded as a new possible pathological marker of hepatic precancerous lesion. PMID- 14761283 TI - [Interleukin-18 antisense oligodeoxynucleotide promotes hepatocyte regeneration of partial liver allograft]. AB - OBJECTIVE: To study the effect of interleukin (IL)-18 ASPODN on regeneration of allogeneic partial liver graft in rats. METHODS: Ninety donor SD rats and ninety recipient LEW rats were randomly divided into 3 groups: 50% partial liver transplantation group (PLT group); PLT+IL-18 antisense phosphorothioate oligodeoxynucleotide (ASPODN) treatment group (IL-18 ASPODN group) and PLT+IL-18 SPODN treatment group (IL-18 SPODN group) in which liposomes encapsulated IL-18 ASPODN or IL-18 SPODN were intravenous injection every day after PLT. BrdU labeling of hepatocytes, expression of IL-18 protein and IFN-gamma mRNA in liver graft, and serum level of IFN-gamma were measured with immunohistochemistry analysis, Western blotting, semi-quantification RT-PCR, and ELISA, respectively. RESULTS: Although regeneration of liver graft from each group peaked 72 hour after transplantation, BrdU labeling of hepatocytes in IL-18 ASPODN group (58.3%+/-7.5%) were significantly higher than those of PLT group (31.6%+/-6.7%) (t=6.503, P<0.001) and IL-18 SPODN group (33.4%+/-5.5%) (t=6.558, P<0.001). Expression of IL-18 protein and IFN-gamma mRNA in liver graft, and serum level of IFN-gamma in IL-18 ASPODN group from 48 hour, 72 hour and 96 hour after transplantation were significantly suppressed compared with PLT group (IL 18protein: t=2.950, t=5.916, t=7.947, P<0.05, P<0.001; INF-gamma mRNA: t=2.558, t=6.292, t=8.925, P<0.05, P<0.001; IFN-gamma level: t=16.998, t=15.483, t=54.723, P<0.001) and IL-18 SPODN group (IL-18 protein: t=2.845, t=6.062, t=6.973, P<0.05, P<0.001; INF-gamma mRNA: t=3.117, t=6.154, t=8.738, P<0.05, P<0.001; IFN-gamma level: t=14.531, t=18.139, t=46.924, P<0.001). CONCLUSION: IL-18 ASPODN could promote hepatocyte regeneration of allogeneic partial liver graft by the suppression of IL-18 and IFN-gamma production. PMID- 14761284 TI - [Relationship between hepatitis B virus markers and quantitative HBV DNA in serum]. PMID- 14761285 TI - [Effect of T-cell vaccination in murine experimental autoimmune hepatitis]. AB - OBJECTIVE: To investigate the effect of T-cell vaccination in murine experimental autoimmune hepatitis (EAH). METHODS: To induce the EAH model, the syngeneic S-100 antigen emulsified in complete Freud's adjuvant was injected intraperitoneally to C57Bl/6 at day 1 and day 7. For T-cell vaccination, splenocytes were removed from animal 2 weeks after induction of EAH and from control animals, and activated in vitro by mitogen stimulation with Concanavalin A (Con A), then inactivated by mitomycin and injected at 5 10(7) cells per animal as T-cell vaccination at 14 and 7 days before first induction of EAH. RESULTS: The histological grade and serum ALT level of the mice who received T-cell vaccination were decrease significantly, compared with that of model group (1.44+/-0.88 vs. 2.33+/-0.87, t=2.24, P<0.05; 63.0U/L+/-23.4U/L vs. 115.0U/L1+/-39.6U/L, t=2.37, P<0.01, respectively); there was no significant change in mice who received irrelevant T cell vaccination. CONCLUSION: T-cell vaccination with T cells from EAH animals, but not with irrelevant T cells, was able to protect animals from EAH. PMID- 14761286 TI - [Experience of prophylaxis and treatment of hepatitis B recurrence in liver transplantation recipients]. PMID- 14761287 TI - [The effect of Hepatitis C virus nonstructural protein 5A on cell cycling]. PMID- 14761288 TI - [Clinical futures and diagnosis of viral hepatitis E]. PMID- 14761289 TI - [Evaluate of the therapeutic efficiency of the Chinese traditional medicine Gansu granule on chronic hepatitis B]. PMID- 14761290 TI - [Clinical value of the histological quantative method of hepatic fibrosis]. PMID- 14761291 TI - [Expression of peroxisome proliferator-activated receptor alpha in human normal liver and hepatocellular carcinoma]. PMID- 14761292 TI - [An improved method of isolation and purification for newborn rat's hepatocytes and the study for preservation and revivfication]. PMID- 14761293 TI - [The expression and their significance of epidermal growth factor, transforming growth factor alpha and epidermal growth factor receptor during the intrahepatic cholangiocarcinoma carcinogenesis]. PMID- 14761294 TI - [Effects of Oxymatrine on the expression of tissue inhibitor of metalloproteinase 1 and alpha-smooth muscle actin in the livers of rats with hepatic fibrosis]. PMID- 14761295 TI - [Confocal laser microscopy: a new method to quantitative analysis the liver tissue type I, III collagen]. PMID- 14761296 TI - [The current progress of autoimmune hepatitis]. PMID- 14761297 TI - [Adefovir dipivoxil-A new drug for hepatitis B]. PMID- 14761298 TI - Quantitative analysis of nucleic acids--the last few years of progress. AB - DNA and RNA quantifications are widely used in biological and biomedical research. In the last ten years, many technologies have been developed to enable automated and high-throughput analyses. In this review, we first give a brief overview of how DNA and RNA quantifications are carried out. Then, five technologies (microarrays, SAGE, differential display, real time PCR and real competitive PCR) are introduced, with an emphasis on how these technologies can be applied and what their limitations are. The technologies are also evaluated in terms of a few key aspects of nucleic acids quantification such as accuracy, sensitivity, specificity, cost and throughput. PMID- 14761299 TI - Global genetic analysis. AB - The introduction of molecular markers in genetic analysis has revolutionized medicine. These molecular markers are genetic variations associated with a predisposition to common diseases and individual variations in drug responses. Identification and genotyping a vast number of genetic polymorphisms in large populations are increasingly important for disease gene identification, pharmacogenetics and population-based studies. Among variations being analyzed, single nucleotide polymorphisms seem to be most useful in large-scale genetic analysis. This review discusses approaches for genetic analysis, use of different markers, and emerging technologies for large-scale genetic analysis where millions of genotyping need to be performed. PMID- 14761300 TI - Structure-based functional discovery of proteins: structural proteomics. AB - The discovery of biochemical and cellular functions of unannotated gene products begins with a database search of proteins with structure/sequence homologues based on known genes. Very recently, a number of frontier groups in structural biology proposed a new paradigm to predict biological functions of an unknown protein on the basis of its three-dimensional structure on a genomic scale. Structural proteomics (genomics), a research area for structure-based functional discovery, aims to complete the protein-folding universe of all gene products in a cell. It would lead us to a complete understanding of a living organism from protein structure. Two major complementary experimental techniques, X-ray crystallography and NMR spectroscopy, combined with recently developed high throughput methods have played a central role in structural proteomics research; however, an integration of these methodologies together with comparative modeling and electron microscopy would speed up the goal for completing a full dictionary of protein folding space in the near future. PMID- 14761301 TI - Post-translational modifications and their biological functions: proteomic analysis and systematic approaches. AB - Recently produced information on post-translational modifications makes it possible to interpret their biological regulation with new insights. Various protein modifications finely tune the cellular functions of each protein. Understanding the relationship between post-translational modifications and functional changes ("post-translatomics") is another enormous project, not unlike the human genome project. Proteomics, combined with separation technology and mass spectrometry, makes it possible to dissect and characterize the individual parts of post-translational modifications and provide a systemic analysis. Systemic analysis of post-translational modifications in various signaling pathways has been applied to illustrate the kinetics of modifications. Availability will advance new technologies that improve sensitivity and peptide coverage. The progress of "post-translatomics", novel analytical technologies that are rapidly emerging, offer a great potential for determining the details of the modification sites. PMID- 14761302 TI - Protein-protein interaction networks: from interactions to networks. AB - The goal of interaction proteomics that studies the protein-protein interactions of all expressed proteins is to understand biological processes that are strictly regulated by these interactions. The availability of entire genome sequences of many organisms and high-throughput analysis tools has led scientists to study the entire proteome (Pandey and Mann, 2000). There are various high-throughput methods for detecting protein interactions such as yeast two-hybrid approach and mass spectrometry to produce vast amounts of data that can be utilized to decipher protein functions in complicated biological networks. In this review, we discuss recent developments in analytical methods for large-scale protein interactions and the future direction of interaction proteomics. PMID- 14761303 TI - Chemical genomics and medicinal systems biology: chemical control of genomic networks in human systems biology for innovative medicine. AB - With advances in determining the entire DNA sequence of the human genome, it is now critical to systematically identify the function of a number of genes in the human genome. These biological challenges, especially those in human diseases, should be addressed in human cells in which conventional (e.g. genetic) approaches have been extremely difficult to implement. To overcome this, several approaches have been initiated. This review will focus on the development of a novel "chemical genetic/genomic approach" that uses small molecules to "probe and identify" the function of genes in specific biological processes or pathways in human cells. Due to the close relationship of small molecules with drugs, these systematic and integrative studies will lead to the "medicinal systems biology approach" which is critical to "formulate and modulate" complex biological (disease) networks by small molecules (drugs) in human bio-systems. PMID- 14761304 TI - Nutriproteomics: identifying the molecular targets of nutritive and non-nutritive components of the diet. AB - The study of whole patterns of changes in protein expression and their modifications, or proteomics, presents both technological advances as well as formidable challenges to biological researchers. Nutrition research and the food sciences in general will be strongly influenced by the new knowledge generated by the proteomics approach. This review examines the different aspects of proteomics technologies, while emphasizing the value of consideration of "traditional" aspects of protein separation. These include the choice of the cell, the subcellular fraction, and the isolation and purification of the relevant protein fraction (if known) by protein chromatographic procedures. Qualitative and quantitative analyses of proteins and their peptides formed by proteolytic hydrolysis have been substantially enhanced by the development of mass spectrometry technologies in combination with nanoscale fluidics analysis. These are described, as are the pros and cons of each method in current use. PMID- 14761305 TI - Bioinformatics in the post-genome era. AB - Recent years saw a dramatic increase in genomic and proteomic data in public archives. Now with the complete genome sequences of human and other species in hand, detailed analyses of the genome sequences will undoubtedly improve our understanding of biological systems and at the same time require sophisticated bioinformatic tools. Here we review what computational challenges are ahead and what are the new exciting developments in this exciting field. PMID- 14761306 TI - From the sequence to cell modeling: comprehensive functional genomics in Escherichia coli. AB - As a result of the enormous amount of information that has been collected with E. coli over the past half century (e.g. genome sequence, mutant phenotypes, metabolic and regulatory networks, etc.), we now have detailed knowledge about gene regulation, protein activity, several hundred enzyme reactions, metabolic pathways, macromolecular machines, and regulatory interactions for this model organism. However, understanding how all these processes interact to form a living cell will require further characterization, quantification, data integration, and mathematical modeling, systems biology. No organism can rival E. coli with respect to the amount of available basic information and experimental tractability for the technologies needed for this undertaking. A focused, systematic effort to understand the E. coli cell will accelerate the development of new post-genomic technologies, including both experimental and computational tools. It will also lead to new technologies that will be applicable to other organisms, from microbes to plants, animals, and humans. E. coli is not only the best studied free-living model organism, but is also an extensively used microbe for industrial applications, especially for the production of small molecules of interest. It is an excellent representative of Gram-negative commensal bacteria. E. coli may represent a perfect model organism for systems biology that is aimed at elucidating both its free-living and commensal life-styles, which should open the door to whole-cell modeling and simulation. PMID- 14761307 TI - Yeast as a touchstone in post-genomic research: strategies for integrative analysis in functional genomics. AB - The new complexity arising from the genome sequencing projects requires new comprehensive post-genomic strategies: advanced studies in regulatory mechanisms, application of new high-throughput technologies at a genome-wide scale, at the different levels of cellular complexity (genome, transcriptome, proteome and metabolome), efficient analysis of the results, and application of new bioinformatic methods in an integrative or systems biology perspective. This can be accomplished in studies with model organisms under controlled conditions. In this review a perspective of the favourable characteristics of yeast as a touchstone model in post-genomic research is presented. The state-of-the art, latest advances in the field and bottlenecks, new strategies, new regulatory mechanisms, applications (patents) and high-throughput technologies, most of them being developed and validated in yeast, are presented. The optimal characteristics of yeast as a well-defined system for comprehensive studies under controlled conditions makes it a perfect model to be used in integrative, "systems biology" studies to get new insights into the mechanisms of regulation (regulatory networks) responsible of specific phenotypes under particular environmental conditions, to be applied to more complex organisms (e.g. plants, human). PMID- 14761309 TI - Transient receptor potential ion channels and animal sensation: lessons from Drosophila functional research. AB - Ion channels of the transient receptor potential (TRP) superfamily are non selective cationic channels with six transmembrane domains. The TRP channel made its first debut as a light-gated Ca2+ channel in Drosophila. Recently, research on animal sensation in Drosophila disclosed other members of the TRP family that are required for touch sensation and hearing as well as the sensation of painful stimuli. PMID- 14761308 TI - Functional genomic approaches using the nematode Caenorhabditis elegans as a model system. AB - Since the completion of the genome project of the nematode C. elegans in 1998, functional genomic approaches have been applied to elucidate the gene and protein networks in this model organism. The recent completion of the whole genome of C. briggsae, a close sister species of C. elegans, now makes it possible to employ the comparative genomic approaches for identifying regulatory mechanisms that are conserved in these species and to make more precise annotation of the predicted genes. RNA interference (RNAi) screenings in C. elegans have been performed to screen the whole genome for the genes whose mutations give rise to specific phenotypes of interest. RNAi screens can also be used to identify genes that act genetically together with a gene of interest. Microarray experiments have been very useful in identifying genes that exhibit co-regulated expression profiles in given genetic or environmental conditions. Proteomic approaches also can be applied to the nematode, just as in other species whose genomes are known. With all these functional genomic tools, genetics will still remain an important tool for gene function studies in the post genome era. New breakthroughs in C. elegans biology, such as establishing a feasible gene knockout method, immortalized cell lines, or identifying viruses that can be used as vectors for introducing exogenous gene constructs into the worms, will augment the usage of this small organism for genome-wide biology. PMID- 14761310 TI - Functional genomics approach using mice. AB - The rapid development and characterization of the mouse genome sequence, coupled with comparative sequence analysis of human, has been paralleled by a reinforced enthusiasm for mouse functional genomics. The way to uncover the in vivo function of genes is to analyze the phenotypes of the mutant animals. From this standpoint, the mouse is a suitable and valuable model organism in the studies of functional genomics. Therefore, there have been enormous efforts to enrich the list of the mutant mice. Such a trend emphasizes the random mutagenesis, including ENU mutagenesis and gene-trap mutagenesis, to obtain a large stock of mutant mice. However, since various mutant alleles are needed to precisely characterize the role of a gene in vivo, mutations should be designed. The simplicity and utility of transgenic technology can satisfy this demand. The combination of RNA interference with transgenic technology will provide more opportunities for researchers. Nevertheless, gene targeting can solely define the in vivo function of a gene without a doubt. Thus, transgenesis and gene targeting will be the major strategies in the field of functional genomics. PMID- 14761311 TI - Proteomic studies in plants. AB - Proteomics is a leading technology for the high-throughput analysis of proteins on a genome-wide scale. With the completion of genome sequencing projects and the development of analytical methods for protein characterization, proteomics has become a major field of functional genomics. The initial objective of proteomics was the large-scale identification of all protein species in a cell or tissue. The applications are currently being extended to analyze various functional aspects of proteins such as post-translational modifications, protein-protein interactions, activities and structures. Whereas the proteomics research is quite advanced in animals and yeast as well as Escherichia coli, plant proteomics is only at the initial phase. Major studies of plant proteomics have been reported on subcellular proteomes and protein complexes (e.g. proteins in the plasma membranes, chloroplasts, mitochondria and nuclei). Here several plant proteomics studies will be presented, followed by a recent work using multidimensional protein identification technology (MudPIT). PMID- 14761312 TI - [Raise the level of studies on childhood epilepsy in China]. PMID- 14761313 TI - [Clinical features and diagnostic points of influenza in children]. PMID- 14761314 TI - [Understanding the hemorrhage due to late onset vitamin K deficiency]. PMID- 14761315 TI - [Characteristics of clinical manifestations and EEG of Lennox-Gastaut syndrome]. AB - OBJECTIVE: Lennox-Gastaut syndrome (LGS) is one of the most severe and refractory form of childhood epilepsy. The purpose of this study was to investigate the clinical and EEG characteristics of patients with LGS. METHODS: Sixty-two patients with LGS, including 37 males and 25 females, were followed-up regularly per three months or per six months, therapy was adjusted according to the changes in seizures and EEG, and the clinical data were analyzed in detail. RESULTS: The onset occurred between the age of 8 months and 12 years, with the peak at 1-4 years of age, accounting for 61%; a late onset which occurred after 8 years of age, was unusual. Furthermore, one patient who developed LGS at the age of 13 years and remained to have all the features of seizures and EEG at 35 years of age was identified as adult's LGS. Forty-three patients were classified as symptomatic, perinatal events were the predominant factors in this group. The others were cryptogenic. It was noted that 11 cases had a history of West syndrome. A transformation process from West syndrome to LGS was observed in another 7 cases. Every patient had two or more seizure types during the course of the disease; tonic seizure, atypical absence seizure, head drop or sudden falls were the characteristic types. The degree of mental deficit was variable from slight to profound deterioration, but mental and behavioral disturbances existed in every case as a rule. In all cases electroencephalogram (EEG) background was abnormal and consisted of diffuse slow spike-and-waves (1-1.5CPS), predominant in frontal and temporal regions. Twenty-four cases had the polyspike-wave. Bursts of fast rhythms (10-14CPS) were observed in 29 patients during sleep. The choice of antiepileptic drugs (AEDs) was based on the seizure types; routinely, 2 or more kinds of AEDs were used in combination, the classic drugs, valproate and clonazepam were firstly recommended; the other drugs, such as lamotrigine and topiramate that are used as add-on therapy were further consideration. Although the total effect was not satisfactory, the severity and frequency of seizures in almost all cases had lessened to some extent. CONCLUSION: LGS shows diverse manifestations; comprehensive diagnosis is crucial, active and efficacious treatment can improve the mental and behavioral development and prognosis as a whole. PMID- 14761316 TI - [Clinical and sleep EEG monitoring characteristics and long-term follow-up study on narcolepsy]. AB - OBJECTIVE: Narcolepsy is a sleep disorder characterized by excessive daytime sleepiness, cataplexy, hypnagogic hallucination and sleep paralysis, with abnormal characteristics of shorter rapid eye movement (REM) sleep latency. The management of the patients is very important. The present study focused on the clinical characteristics, diagnostic methods and long-term prognosis of this particular syndrome. METHODS: The clinical data of 39 narcoleptic children were analyzed. Sleep EEG monitoring was performed in all patients. Among the 39 cases, 23 were followed up. RESULTS: All the patients manifested with excessive daytime sleepiness, with disrupted nocturnal sleep occurring in 35 cases. Cataplexy appeared in 36 cases, and sleep paralysis in 9, hypnagogic hallucination in 19, and automatic behavior in 6 cases, respectively. Sleep EEG monitoring demonstrated a short mean sleep latency (< 5 minutes) and two or more sleep onset REM periods (SOREMPs) in 38 cases. Twenty-three of the 39 cases were followed-up. Seventeen cases were followed-up for over one year. The longest follow-up duration was 14 years. Methylphenidate was administered in 10 cases. The excessive daytime sleepiness had been improved in 7 cases (70%). No obvious adverse effects were found. Psychosocial and academic problems appeared in most cases. CONCLUSION: Narcolepsy is a chronic neurological disorder. A definite diagnosis is established when the symptoms of cataplexy and excessive daytime sleepiness occur in association with the characteristic findings on sleep EEG monitoring. Appropriate drug therapy and psychosocial management are of help for such patients. Stimulant medication is an important component of the overall treatment program. A comprehensive approach is necessary to meet the needs of children with narcolepsy. Family education and emotional support are key elements in the management plan. The overall goal for managing childhood narcolepsy is to assist the child and family in achieving optimal quality of life. PMID- 14761317 TI - [Clinical observation and long-term follow-up of benign infantile epilepsy]. AB - OBJECTIVE: To investigate clinical characteristics, EEG changes and therapeutic response of benign infantile epilepsy and to study the early diagnostic methods. METHODS: Clinical observation and Video-EEG monitoring were carried out in babies with convulsions at 3 - 24 months of age. In these children, febrile convulsion, symptomatic epilepsies and developmental abnormalities were excluded, and the therapeutic effect and long-term outcome were followed up. RESULTS: Forty-two babies were diagnosed to have benign infantile epilepsy by two-year follow-up. Three of them had familial history of benign infantile convulsions. Nineteen percent had mild diarrhea during the onset of convulsions, cluster seizures occurred during a short period in 67% of cases and no status epilepticus occurred. Video-EEG monitoring confirmed seizures originating from temporal, occipital or multifocal areas separately in 3 patients with partial seizures. Interictal EEG background was normal and there were Rolandic small spikes during sleep in 24% of patients. Thirty-nine patients were treated with single antiepileptic drugs and the mean treatment course was 9 months. Three cases did not take medicine. All the patients were seizure free within a year. CONCLUSION: Benign infantile epilepsy should be considered when the following characteristics occur in early stage of the disease: (1) convulsions occurring between 3 to 12 month of age and not later than 24 months of age with or without familial history of benign infantile convulsion; (2) normal psychomotor development before and after convulsion occurs; (3) no evoked factors or only mild diarrhea; (4) majority of cases have partial seizures, or secondary generalized seizures. There are often cluster convulsions during the onset stage, but no status epilepticus; (5) normal EEG background and there may be Rolandic small spikes during sleep; (6) normal neuroimaging. PMID- 14761318 TI - [Malnutrition increases hippocampal neurogenesis in the immature rat after status epilepticus]. AB - OBJECTIVE: Neurogenesis in the dentate gyrus of hippocampus persists in brain of the immature and adult mammalian including human and it can be regulated by physiological and pathological events including nutritional status and seizures. The present study was designed to investigate the potential effects of malnutrition followed by status epileptics on hippocampal neurogenesis in the immature rat. METHODS: Rat pups were divided into 4 groups: malnourished (M), nourished (N), malnourished plus seizures (MS) and nourished plus seizures (NS). The rat pups of group M and group MS were maintained on a starvation regimen from postnatal day 2 (P2) to P18. The status epilepticus of the rat pups in group MS and group NS was elicited by unilateral microinfusion of kainic acid (KA) into the amygdula at P15. Rat pups of the 4 groups were given bromodeoxyuridine (BrdU) intraperitoneally twice daily for 2 days beginning at P17. At P19, the rat pups were killed and the brains were processed for BrdU mitotic labeling combined with double-label immunohistochemistry using early neuron- or glia-specific markers TuJ1 (beta III tubulin) or GFAP (glial fibrillary acidic protein). RESULTS: There were no significant differences in the latent time of seizure between group M and group N [(12.4 +/- 2.6) min vs. (12.1 +/- 2.9) min, P < 0.05]. Histological assessment did not reveal any evidence of hippocampal cell loss after status epilepticus in either group. BrdU-labeled cells were significantly higher in the rats of group MS (374 +/- 18) than group M (303 +/- 20), group NS (312 +/- 24) than group N (269 +/- 18), respectively (P < 0.01). There was also significant difference between group M and group N, group MS and group NS, respectively (P < 0.01). No significant difference was seen between the rats of group NS and group M (P > 0.05). Approximately 60% of BrdU-labeled cells coexpressed TuJ1, and 5% approximately 10% of those co-expressed GFAP. CONCLUSION: Early malnutrition do not alter KA seizure susceptibility and the behavioral manifestations of seizures at P15. Although malnutrition and status epilepticus can increase the proliferation of newly developed cells in the immature rat respectively, malnutrition followed by status epilepticus further increases this proliferation. Furthermore, most of newly developed cells differentiate into early neurons. PMID- 14761319 TI - [Effects of recurrent audiogenic seizures on hippocampal structure and seizure behavior of P77PMC rats]. AB - OBJECTIVE: To investigate the functional role of hippocampal mossy fiber sprouting in the pathophysiologic mechanism of initiation and propagation of epilepsy. METHODS: The authors examined hippocampal mossy fiber synaptic reorganization and the changes of hippocampal neurons in P77PMC rats at different stages in the course of recurrent seizures using Timm's method of silver sulfide staining and Nissl staining and observed the effects of recurrent audiogenic seizures (AGSs) on seizure behavior of P77PMC rats. RESULTS: Frequent recurrent AGSs could cause neuronal loss in CA(1) region of hippocampus and hippocampal mossy fiber sprouting got into the inner molecular layer of dentate gyrus in P77PMC rats, and could decrease the latency of IV/V grade of AGSs, increase the durations of AGSs. The mean A of CA(1) region of hippocampus in Nissl staining after 50 times of AGSs was 35.3 +/- 0.8, which was markedly lower than that of the control (44.1 +/- 0.5; F = 333.89, P < 0.001). The mean A of the inner molecular layer of dentate gyrus in Timm's staining after 50 times of AGSs was 49.3 +/- 4.6, which was markedly higher than that of the control (26.8 +/- 1.7; F = 76.83, P < 0.001). After 30 and 50 times of AGSs, the latent periods of IV/V grade of AGSs were 12 +/- 8 (t = 3.805; P < 0.02) and 17 +/- 7 (t = 5.927; P < 0.002) seconds shorter than the initial period of stimulation respectively on average, and the durations of AGSs were 19 +/- 18 (t = 2.644; P < 0.05) and 10 +/ 7 (t = 3.780; P < 0.02) seconds longer. CONCLUSION: Hippocampal mossy fiber sprouting and neuronal loss not only presents in limbic seizure, but also in AGS, the seizure can be initiated in brainstem but rapidly generalized;in AGS-prone rats, recurrent AGSs can cause mossy fiber synaptic reorganization and neuronal loss in hippocampus, and can also enhance seizure susceptibility of P77PMC rats. In the course of recurrent AGSs, enhanced seizure susceptibility happened before hippocampal mossy fiber sprouting. Their temporal relationships indicate that the anatomical changes may be preceded by functional changes of elevated excitability in the brain. PMID- 14761321 TI - [Cortical neuron injury occurs following recurrent epileptiform discharges induced by magnesium-free treatment in vitro]. AB - OBJECTIVE: To study cortical neuron injury following recurrent epileptiform discharges induced by magnesium-free treatment in vitro. METHODS: Cultured embryo cortical neurons were exposed to magnesium-free media for 3 h, then they were returned to regular media containing normal level magnesium. At different time after Mg(2+)-free treatment, trypan blue staining and determination of LDH activity were used to determine the cell viability, flow cytometry was applied to measure neuronal apoptosis, and MTT assay to study metabolic rate. RESULTS: (1) Neuronal morphology on light microscopy following Mg(2+)-free treatment showed that there were no prominent alterations. (2) At different time (6, 12, 72 h) after Mg(2+)-free treatment, neuronal viability by trypan blue staining and LDH activity showed modest changes compared with time-matched control in different culture days (6, 12, 17 d) (P > 0.05). (3) Cell apoptosis increased mildly at different time after Mg(2+)-free treatment in neurons cultured for different days, but the increase was not significant (P > 0.05). (4) Metabolic rate decreased at 6 h after Mg(2+)-free treatment (P < 0.05) in neurons cultured for 6 d, and was 86.4% of that of the control; while the rate at 24 h in neurons cultured for 12 d and 17 d also decreased (P < 0.05), being 78.7% and 70.9%, respectively, of that of the control. CONCLUSIONS: These findings demonstrated that the injury occurred on cultured cortical neurons caused by magnesium-free treatment-induced recurrent epileptiform discharges was mainly functional and relatively mature neurons displayed more severe and much later mitochondrial function impairment than immature neurons. PMID- 14761323 TI - [Clinical manifestations of 30 cases with Lyme disease]. AB - OBJECTIVE: To understand the clinical characteristics of Lyme disease in children. METHODS: Case records of 30 children with Lyme disease who were admitted to the pediatric department of Forest Hospital, Inner Mongolia from May 1995 to Oct. 2000, were reviewed. Twenty-one cases were male and 9 were female. Their age ranged from 2 to 14 years. The possible factors associated with the onset, incubation period, clinical manifestations of the organ systems and response to treatment were analyzed. RESULTS: Each of the 30 cases had a history of tick bite for one time or more. The incubation period was 1 to 14 days. All of them were hospitalized for typical erythema migrans. Among the 30 cases, 21 (70%) showed symptoms of acute upper respiratory tract infection; 4 (13%) showed circulatory system symptom and manifestations of myocardial lesion; 3 (10%) had joint pain; 1 case (3%) had eye symptom and another case (3%) had nervous system symptoms, such as headache, vomiting, drowsiness, neck stiffness and so on. EEG and spinal fluid test showed some abnormal results. All the 30 cases were positive for IgG antibody to Lyme spirochetes with titers higher than 1:128. Penicillin or cefazolin was effective in treatment of the patients. CONCLUSION: The incubation period of the disease observed among this series of patients was 1 14 d. Lyme disease in this group of patients involved multiple organ systems while erythema migrans and arthralgia were the most frequently seen manifestations. Most of the patients had a favorable outcome with no sequelae. PMID- 14761324 TI - [Clinical characteristics of obstructive sleep apnea syndrome in children]. AB - OBJECTIVE: To explore the clinical features and diagnosis of obstructive sleep apnea syndrome (OSAS) in children. METHODS: Sixty children with OSAS were reviewed, every patient was monitored with polysomnography (PSG) for 7 hours at night for 11 parameters, including the longest apnea time (LAT), apnea and hypopnea index (AHI), the lowest oxygen saturation (SaO(2)), and snore index etc., the parameters of the 2 groups were compared. Meanwhile, tonsillectomy and adenoidectomy were performed for 40 cases of OSAS, and the parameters obtained before and after operation were analyzed. RESULTS: Adenotonsillar hypertrophy and the loud snoring during sleep were found in all cases. The mean values of the PSG parameters were as follows: the longest apnea time was 53 (8-178) seconds (s); the total time of apnea was 310.5 (26-5,260) s; the time of apnea was 26 (3-240) s; the longest hypopnea time was 41 (5-94) s; the total times of hypopnea was 170 (5-2,860) s; the time of hypopnea was 10 (1-85); the apnea index was 4.1 (0.5 25.9); the hypopnea index was 1.4 (0-16.1); the apnea and hypopnea index (AHI) was 6.8 (0.5-38.2); the snore index was 81.7 (1.3-414.8); the lowest saturation of oxygen was 0.78 (0.25-0.93). There was not statistically significant difference in the parameters between 2-7 year group and over 7 year group (P > 0.05). The parameters of postoperation group: the mean value of the longest apnea time was 15.5 (0-60) s; the total time of apnea was 56.4 (60-205) s; the time of apnea was 10.33 (0-40); the longest hypopnea time was 13.25 (0-30) s; the total times of hypopnea was 44.6 (0-73); the hypopnea time was 4.32 (0 - 30) s; the apnea index was 0.6 (0-12); the hypopnea index was 0.62 (0-4); the apnea and hypopnea index (AHI) was 1.25 (0.1-12); the snore index was 30.08 (1.8-102); the lowest oxygen saturation was 93.5% (64%-97%). Compared with preoperation groups there was a statistically significant difference (P < 0.01). Clinically effective rate of the surgeries was over 90%. CONCLUSION: Adenotonsillar hypertrophy seemed to be an important cause of OSAS in children. Snoring, dyspnea, apnea and low ventilation are the major clinical characteristics of OSAS in children. Confirmed diagnosis of the syndrome in children requires PSG recordings. PMID- 14761325 TI - [Genotype and phenotype correlation in Chinese patients with Wilson's Disease]. AB - OBJECTIVE: To determine distribution and mutation pattern of type P ATP7B gene mutation and to explore genotype and phenotype correlation in patients with Wilson's disease (WD). METHODS: Sixty patients with WD from 57 no kinship families, 37 male and 23 female, were enrolled in this study. The age of onset ranged from 4.6 - 39 years, < or = 16 years in 55 patients. Some exons of ATP7B gene mutation were analyzed in patients with WD by using biochemical methods, polymerase chain reaction-single strand configuration polymorphism (PCR-SSCP), restriction fragment and DNA sequence analysis. Totally 778 coding regions were identified with restriction enzyme Msp I. The activity of Cu-ATPase was assessed by measuring inorganic phosphorus in 3 patients with known genotype. RESULTS: Fifty-two of 60 patients (86%) had presented with hepatic manifestations, 30 of them had only hepatic manifestations, 12/52 patients had hepatic and neurological manifestations at the same time; 10/52 patients had hepatic and other symptom; 7/60 patients had only neurological symptom, one patient had no symptom. Eleven mutations were detected by DNA sequencing, including five missense mutations (R778L, V1140A,G943S, V1106I and V1216M), one deletion (1384del17) and five polymorphisms (IVS4-5T/C, A2495G, C2310G, IVS18 + 6C/T and IVS20 + 5A/G) were identified. R778L mutation was identified 52/114 alleles (45.6%). R778L occurred in 38/52 patients with hepatic manifestation (73%), homozygosis of R778L was demonstrated in 14 patients and heterozygosity of R778L in 24 patients. V1106I mutation was 1.7%, G943S, V1140A, and V1216M was 0.86% respectively in this study. Two patients with delayed onset of neurological symptoms occurred V1106I mutation of ATP7B. Cu-ATPase activity of 3 patients with known mutation (R778L/V1106I, R778L/V1216M and R778L/R778L) declined by 44.55%, 88.23% and 69.49%, respectively, compared with normal control. CONCLUSION: The 1384del17bp and V1106I are two novel mutations found in patients with WD. R778L was common mutation of ATP7B gene with frequency of 45.6% in this study. The mutation in exon 8 of WD gene may play an important role in pathogenesis of WD in Chinese. Carriage of R778L mutation seems to be correlated with hepatic manifestation. PMID- 14761326 TI - [Resting energy expenditure in healthy newborn infants]. AB - OBJECTIVE: To establish the value of the resting energy expenditure (REE) in healthy newborns and evaluate relative factors of REE. METHODS: One hundred and fifty-four healthy newborns (75 boys, 79 girls; birth-weight 2,500-3,999 g) were enrolled in this study. The Apgar score at the 5th minute was equal to or more than 8; the postnatal age was equal to or more than 5 days. The newborns had no apparent defect. The mothers had no history of metabolic and endocrine diseases. REE was measured by Deltatrac II in child canopy mode for 30-45 minutes during asleep or quiet awake status. RESULTS: The average REE was (201.8 +/- 25.4) kJ/(kg.d), which was significantly lower than the predicted REE by Schofield formula[(226.1 +/- 4.8) kJ/(kg.d), P = 0.000], the predicted REE was 12.04% higher than the measured REE. There were no differences in sex and ways of delivery. The newborns whose birth-weight was between 2,500-2,999 g were measured in two modes: baby and child mode, and the REE values were significantly higher (122.6 +/- 25.0) kJ/(kg.d) and (208.8 +/- 26.4) kJ/(kg.d), respectively (P = 0.000). CONCLUSIONS: The prediction formula might be improper for calculating the REE in newborn infants. The indirect calorimetry was the better way to know the actual REE of newborns. The authors recommended that in child mode the measurement of REE in newborns would be the indirect calorimetry, and the REE in healthy newborns was (201.8 +/- 25.4) kJ/(kg.d). PMID- 14761327 TI - [Huperzine A attenuates cognitive deficits and brain injury after hypoxia ischemic brain damage in neonatal rats]. AB - OBJECTIVE: To investigate the protective effects of Huperzine A, a potent acetylcholinesterase inhibitor, against the hypoxic ischemic brain damage (HIBD) of the cognitive and morphology in the neonatal rats. METHODS: Postnatal 7 days old rats were given vehicle or Huperzine A (0.05 mg/kg or 0.1 mg/kg, i.p.) following HIBD (unilateral carotid artery ligation followed by hypoxia) or sham operation, and then tested the learning ability and memory in the Morris water maze (MWM) from 36 to 40 postnatal days. The performance in MWM (escape latency, probe time) were recorded to evaluate the learning and memory dysfunction. At the end of MWM trials, the rats were decapitated and their brains were histologically analyzed. The tissue loss in different brain regions including striatum, cortex, and hippocampus were analyzed by image analysis system. The CA(1) subfield neurons numbers were counted to evaluate the brain damage. The acetylcholinesterase histochemistry staining was used to determine the activity of acetylcholinesterase in different brain regions. RESULTS: Compared with sham operated group, HIBD rats with the vehicle treatment displayed significant tissue losses in the hippocampus (including CA(1) neurons), cortex, and striatum, as well as severe spatial memory deficits (escape latency: 44 s vs 30 s, P < 0.05, probe time: 14 s vs 40 s, P < 0.01). Huperzine A treatment (0.1 mg/kg) resulted in significant protection against both HI-induced brain tissue losses and spatial memory impairments (mean escape latency: 34 s vs 44 s, P < 0.05, probe time: 35 s vs 14 s,P < 0.01). However, Huperzine A treatment (0.05 mg/kg) did not show any significant improvement of spatial memory impairments (mean escape latency: 45 s vs 44 s, P > 0.05, probe time: 17 s vs 14 s, P > 0.05), but moderate to severe brain tissue losses. There was a pronounced reduction of CA(1) neuron density in ipsilateral hemisphere of vehicle-treated group and 0.05 mg/kg Huperzine A group compared with contralateral hemisphere or ipsilateral hemisphere of sham-operated group and 0.1 mg/kg Huperzine A group (72 vs 232, P < 0.01, 72 vs 229, P < 0.01, respectively). There was a close linear correlation between the CA(1) neurons cell number and the mean escape latency for 5 d acquisition trials (r = 0.777, P < 0.01). CONCLUSION: The unilateral HI brain injury in a neonatal rat model was associated with cognitive deficits, and that Huperzine A treatment may be protective against both brain injury and spatial memory impairment. Huperzine A showed a therapeutic potential for the treatment of hypoxic-ischemic encephalopathy (HIE) caused by the perinatal asphyxia. PMID- 14761328 TI - [Diagnosis and prognosis of systemic juvenile rheumatoid arthritis]. PMID- 14761329 TI - [Association of intelligence structures in cerebral palsy children and high risk factors]. PMID- 14761330 TI - [Cerebral protection effects of 7-nitroso-indazole on ischemic-hypoxic neonatal rats brain]. PMID- 14761331 TI - [Long term learning memory and long term potential of hippocampus in cerebral injury of ischemic hypoxic neonatal rats]. PMID- 14761332 TI - [Comparison of pulmonary function of asthma children determined with impulse oscillometry and maximal expiratory flow-volume curve]. PMID- 14761333 TI - [Effects of cefathiamidine on the treatment of children with acute bacterial infectious disease]. PMID- 14761334 TI - [Association between gastroduodenal diseases and cagA, vacA gene expressions of Helicobacter pylori]. PMID- 14761335 TI - [Childhood epilepsy with occipital paroxysms of panayiotopoulos type, report of 8 cases]. PMID- 14761336 TI - [Dopa-responsive dystonia in children]. PMID- 14761337 TI - [Application of bronchoalveolar lavage in the treatment of atelectasis caused by extubation after endotracheal intubation]. PMID- 14761338 TI - [Making the scoring card based on Pediatric Critical Illness Score]. PMID- 14761339 TI - [Report of a case with Wilson-Mikity syndrome]. PMID- 14761340 TI - [The 9th International Child Neurology Congress and the 7th Asian and Oceanian Congress of Child Neurology held in Beijing]. PMID- 14761341 TI - [Effects and treatment strategy of epilepsy form discharge on the cortex high function]. PMID- 14761342 TI - [Progresses in clinical studies on treatment of childhood asthma with corticosteroid inhalation]. PMID- 14761343 TI - [Diagnosis, treatment and prevention of anthrax]. PMID- 14761344 TI - [Clinical applications of tidal breathing analysis in infants and young children]. PMID- 14761346 TI - [Toxic effects of ammonium perchlorate on thyroid of rats]. AB - OBJECTIVE: To study the effects of toxicity of ammonium perchlorate (AP) on thyroid of rats. METHODS: Eighty-eight Wistar rats were treated orally with different dosages of AP. Three treated groups received 129, 257, 514 mg.kg(-1).d( 1) of AP respectively and one control group drunk water for 13 weeks. Another 3 groups received 1.2, 46.5, 465.0 mg.kg(-1).d(-1) of AP respectively and one control group drunk water for 36 weeks. The behavior and change of body weight in rats were observed. The levels of thyroid hormones in serum were measured and the pathological changes of thyroid tissue were observed as well. RESULTS: There were no differences in behavior and change of body weight between different AP exposure time. When the rats were treated with AP 514 mg for 13 weeks, free triiodothyronine (FT3, 2.48 pmol/L), free thyroxin (FT4, 13.33 pmol/L) were lower than those in control group (3.24, 20.92 pmol/L respectively, P<0.05). Thyroid stimulating hormone (TSH, 0.375 mIU/L), thyroglobulin (TG, 3.37 microg/L) were higher than those in control group (0.29 mIU/L, 2.00 microg/L respectively, P<0.05). When the rats were treated with AP 465 mg for 36 weeks, FT3 (2.65 pmol/L) was lower than that in control group (4.97 pmol/L, P<0.01). FT4 in 46.5, 465 mg groups (10.63, 2.17 pmol/L respectively) were lower than that in control group (15.74 pmol/L, P<0.05, P<0.01). TSH in 465 mg group (0.34 mIU/L) was higher than that in control group (0.14 mIU/L, P<0.05). Histopathologic examination showed that follicle proliferation, no colloid in follicle, gore, follicular diminishing or atresia were found in 46.5, 465 mg groups with a dose-effect relationship. CONCLUSIONS: The toxic effects of AP on the growth of rats were not found, but those on the thyroid of rats were found significantly. Thyroid is the target organ of AP. It is considered that none effect dose of AP for rat thyroid may be 1.2 mg.kg(-1).d(-1), its threshold dose may be 46.5 mg.kg(-1).d(-1). PMID- 14761347 TI - [Effect of lead acetate on the nerve growth factor protein expression and the regulation of thyroid hormone]. AB - OBJECTIVES: To study the effect of lead acetate on the expression of nerve growth factor (NGF) protein in rat brain and the regulation of thyroid hormone. METHODS: Lead acetate was given to SD rats intraperitoneally ip. at the dosage of 25, 50 and 100 mg/kg respectively. 6-n-propyl-2-thiouracil (PTU) was used to make a hypothyroid model and then lead acetate was given at the dosage of 50 mg/kg body weight through i.p. The NGF protein expression in rat brain was observed by immunohistochemistry Triiodothyronine (T3), thyroxin (T4), TSH in serum and T3, T4 in brain tissue were determined by radio immunoassays (RIAs). RESULTS: The average gray value of NGF protein in cerebral cortex of 50 mg, 100 mg treated groups (180.49 +/- 10.33, 169.72 +/- 19.75, respectively) were lower than the control (200.75 +/- 3.27, P<0.01). The area density of NGF protein in hippocampus of three treated groups (0.08 +/- 0.14, 0.12 +/- 0.02, 0.13 +/- 0.04, respectively) were significantly different from the control (0.025 +/- 0.015, P<0.05). The area density and the average gray value of NGF protein in lead acetate treated hypothyroid rat brain were of no significant changes. The levels of serum T3 in three treated groups [(0.68 +/- 0.02), (0.57 +/- 0.04), (0.54 +/- 0.02) microg/L respectively] and T4 [(28.30 +/- 1.83), (27.35 +/- 2.55), (24.00 +/- 3.01) microg/L] in serum were significantly lower while TSH [(6.34 +/- 1.13), (7.74 +/- 0.79), (9.16 +/- 0.77) IU] higher than those in the control [T3 (0.97 +/- 0.14) microg/L, T4 (54.50 +/- 3.70) microg/L and TSH (4.62 +/- 2.16) IU], and there was a good dose-response relationship. The levels of T3 in cerebral cortex of three treated groups [(13.26 +/- 0.81), (11.49 +/- 0.10), (10.42 +/- 1.19) pg/mg pro respectively] and T4 [(0.50 +/- 0.03), (0.49 +/- 0.13), (0.42 +/- 0.01) ng/mg pro] were significantly lower than those in control [(20.85 +/- 11.01) pg/mg pro, (0.76 +/- 0.14) ng/mg pro, P<0.05, P<0.01]. CONCLUSION: Lead could increase the NGF protein expression in rat brain, which may be regulated by thyroid hormone. PMID- 14761348 TI - [The improvement of taurine in learning and memory ability of rats exposed to lead]. AB - OBJECTIVE: To investigate the improvement of taurine (Tau) in learning and memory ability of rats exposed to lead. METHODS: Forty Wistar rats were randomly divided into control group: treated with distilled water; lead group: treated with lead acetate (40 mg.kg(-1).d(-1)); lead-taurine group 1, 2, 3: lead acetate (40 mg.kg( 1).d(-1)) + different concentrations of taurine (100, 400, 800 mg.kg(-1).d(-1)). The ability of learning and memory of rats were measured weekly by spatial water maze test from the 5th to 8th week. At the end of the experiment, the rats were killed, the samples of blood and brain were taken for test. RESULTS: (1) The time of seeking anchorage of lead-Tau 800 mg group in the 6th, 7th, 8th week and that of lead-Tau 400 mg group in the 6th week were significantly lower than that of lead group (P<0.05). (2) Blood lead contents in lead-Tau 100 mg and lead-Tau 400 mg group [(510.9 +/- 57.56) microg/L, (485.40 +/- 98.85) microg/L] were different from those in lead group (P<0.05). (3) The content of malondialdehyde (MDA) and the activities of superoxide dismutase (SOD), nitric oxide synthase (NOS), acetylcholinesterase (AChE) in brain of lead-Tau 800 mg group and lead-Tau 400 mg group were also different from those in lead group (P<0.05 or P<0.01). The content of GSH and the activity of GSH-Px in lead-Tau 800 mg group were different from those in lead group (P<0.05) as well. CONCLUSION: Taurine could improve learning and memory ability of rats exposed to lead and may play a protective role in brain. PMID- 14761350 TI - [Induction of aryl hydrocarbon receptor and CYP1A1 mRNA by 2,3,7,8 tetrachlorodibenzo-p-dioxin in rat liver]. AB - OBJECTIVE: To explore the toxic mechanism of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) by studying the induction of cytochrome P4501A1 (CYP1A1) and aryl hydrocarbon receptor (AHR) mRNA in liver of TCDD-treated SD rats. METHODS: Thirty female SD rats were randomly divided into control group and 5 exposure groups, every group had 5 rats. The animals were treated i.p. with 0.01, 0.1, 1, 10, 50 microg TCDD/kg BW. AHR and CYP1A1 mRNA expression were analyzed by RT-PCR after 24 h. RESULTS: The contents of AHR and CYP1A1 mRNA were increased in all exposure groups except the 0.01 microg TCDD/kg BW group. AHR mRNA content was significantly increased in 50 microg TCDD/kg BW group (P<0.05); CYP1A1 mRNA contents were significantly increased in all exposure groups (P<0.05) but not 0.01 microg TCDD/kg BW group. There were dose-response relationship between TCDD doses and AHR, CYP1A1 gene expression. CONCLUSION: Both AHR and CYP1A1 gene in liver of TCDD-treated SD rats can be induced 24 h after exposure and CYP1A1 gene is more inducible than AHR gene. PMID- 14761351 TI - [Clinical manifestation and ultrasonic characteristics of five patients with acute arsenic poisoning]. AB - OBJECTIVE: To study the clinical manifestation and ultrasonic characteristics of liver, kidney and heart of five patients with acute arsenic poisoning. METHODS: The activity of serum myocardial enzymes, function of liver and kidney, and urinary As concentrations were measured. HDI 3000 Enhanced, and Toshiba 38A two dimensional ultrasound was used to examine the ultrasonic echogram of heart, liver, kidney of the patients. RESULTS: (1) The arsenic concentrations in the urine (1.9 approximately 15.6 micromol/L) were higher than the normal value (1.17 micromol/L) in these patients (blood dialytic fluid of one patient with anuria was measured); (2) Four of them had increased WBC, or anemia, and abnormal urine routine to various degree; (3) The activities of serum myocardial enzymes (CK, AST, LDH and HBDH) in 4 patients were at least 2 items increased; (4) Serum bilirubin and urea nitrogen in all patients were increased; (5) The ultrasonic echogram of liver and kidney in these 5 patients showed abnormality to various degree, one of them had slight enlargements in left atrium and ventricle as well as a little pericardial fluid. CONCLUSION: The clinical manifestation and ultrasonic characteristics of liver, kidney, and heart were consistent with the pathologic changes in acute arsenic poisoning. Early blood dialysis may reduce visceral damage. PMID- 14761352 TI - [DNA damage and changes of antioxidative enzymes in chronic benzene poisoning mice]. AB - OBJECTIVE: To study the damage effect of benzene on DNA and its mechanism and the changes of antioxidative enzymes in vivo. METHODS: DNA break in bone marrow cells and peripheral blood lymphocytes of mice exposed to benzene by 4 h static inhalation per day at different concentrations for two months were analyzed with single cell gel electrophoresis (SCGE). Meanwhile, the activity of SOD, GSH-Px and the level of MDA in liver, spleen and brain were detected. RESULTS: In low and high dosage groups, the rate of DNA migration of bone marrow cells (83.56% +/ 10.28%, 92.54% +/- 15.93%) and peripheral blood lymphocytes (41.27% +/- 6.03%, 65.79% +/- 11.62%) were higher than those in control (4.13% +/- 0.52% and 2.21% +/- 0.31% respectively, P<0.05]. The activity of SOD in liver [(754.33 +/- 116.30), (694.26 +/- 116.30) U/mg pro] and GSH-Px [(22.52 +/- 3.31), (18.56 +/- 4.97) U/mg pro] were lower than those in control [(999.92 +/- 188.24) and (35.31 +/- 6.63) U/mg pro respectively, P<0.05, P<0.01]. But there was no significant difference between the two dosage groups. The activity of GSH-Px in spleen of both groups [(31.38 +/- 2.71), (25.30 +/- 7.44) U/mg pro] were lower than that of control [(37.11 +/- 3.42) U/mg pro, P<0.05] and there was significant difference between the two dosage groups. The activity of GSH-Px in brain of both groups [(5.70 +/- 0.84), (5.24 +/- 1.19) U/mg pro, P<0.05] were lower than that of control [(7.10 +/- 0.46) U/mg pro, P<0.05], but there was no significant difference between the two dosage groups. The level of MDA in brain of high dosage group [(3.99 +/- 1.15) nmol/mg pro] was higher than that of control [(2.58 +/- 0.53) nmol/mg pro, P<0.05]. CONCLUSION: Chronic benzene poisoning may result in DNA break in bone marrow cells and peripheral blood lymphocytes and decrease in the activity of antioxidative enzymes. PMID- 14761353 TI - [Urinary excretion pattern of 2-thio-thiazolidine-4-carboxylic acid in workers exposed to carbon disulfide]. AB - OBJECTIVE: To study the urinary excretion pattern for of 2-thio-thiazolidine-4 carboxylic acid (TTCA) in workers exposed to CS2, so as to provide experimental basis for working out biomonitoring measures for short-term exposure to CS2. METHODS: Sixty-nine subjects were divided into three groups: (1) fourteen volunteers who had not been exposed to CS2 before were exposed to CS2 for 2 hours, their urine samples were collected and analyzed at different time points; (2) The urine samples of 15 occupational exposure workers were collected on pre shift, mid-shift, post-shift; (3) The relationship between 8 h time weighted average CS2 exposure concentrations (PC-TWA) and TTCA levels of post-shift urine was studied among 40 workers chronically exposed to CS2. RESULTS: (1) In the 1st group, urine TTCA level reached the peak [(1.03 +/- 0.72) mg/gCr] 4 h after exposure; (2) In the 2nd group, urine TTCA level on pre-shift [(0.37 +/- 0.28) mg/gCr] was lower than that on mid-shift [(1.23 +/- 0.71) mg/gCr, P<0.01] and post-shift [(1.31 +/- 0.78) mg/gCr, P<0.01]; (3) In the 3rd group, there was a linear relationship between the post-shift urine TTCA level and 8 h CS2 exposure concentrations dose (PC-TWA). The regression equation is Y(TTCA mg/gCr)=1.163 6X(CS2 mg/m3)-5.411 6. CONCLUSION: The post-shift urine TTCA levels may be regarded as a bio-monitoring index for workers exposed to CS2. PMID- 14761354 TI - [An immunohistochemical study of the effects of excessive fluoride on type I collagen in rat developmental dentine]. AB - OBJECTIVE: To study the effects of excessive fluoride on type I collagen in rat developmental dentine. METHODS: Eighty SD rats, 5 days old, were divided into experimental and control groups, 40 in each group. The experimental group received subcutaneous injection of 0.2% NaF every 4 days (the dose was 2 mg NaF per kg body wt). The same volume of 0.9% NaCl was used in the control. Twenty rats in each group were killed 4 days after the second and the seventh injection respectively. The expression of type I collagen was assayed with immunohistochemical technique. RESULTS: 4 out of 20 rats after two injections showed abnormal distribution of type I collagen (dense stain of collagen in the odontoblast, aggregation of collagen in the dentine and disordered arrangement of collagen in the predentine; All 20 rats after seven injections showed abnormal distribution of type I collagen. CONCLUSION: Excessive fluoride may affect the metabolism of type I collagen in rat developmental dentine. PMID- 14761355 TI - [Renal dysfunction in workers exposed to arsenic and cadmium]. AB - OBJECTIVE: To examine the nephrotoxicity induced caused by combined effect of arsenic and cadmium in exposed workers. METHODS: Urinary cadmium and arsenic were used as the exposure biomarkers of cadmium and arsenic. Urinary beta2 microglobulin (Ubeta2-MG), albumin (UALB) and N-acetyl-beta-D-glucosaminidase (UNAG) were measured as the effective biomarkers of tubular and glomerular dysfunction induced by cadmium and arsenic. RESULTS: The combination of cadmium and arsenic induced more severe renal injury than that caused by either of the chemicals given alone. There were positive correlations and significant dose effect among the concentrations of urinary cadmium, arsenic and levels of Ubeta2 MG, UALB, UNAG (P<0.05, P<0.01). CONCLUSION: Cadmium combined with arsenic may have additive effect on renal dysfunction in workers exposed to arsenic and cadmium. PMID- 14761356 TI - [The level of serum tumor necrosis factor-alpha and epidermal growth factor in patients with medicament-like dermatitis by trichloroethylene and its clinical significance]. AB - OBJECTIVE: To investigate the level of tumor necrosis factor alpha (TNF-alpha) and epidermal growth factor (EGF) in patients with medicament-like dermatitis by trichloroethylene (DMLT). METHODS: Using radioimmunoassay methods, serum TNF alpha, EGF were measured in 39 patients with DMLT and in 20 controls. RESULTS: The levels of serum TNF-alpha, EGF in patients with DMLT [(0.278 +/- 0.092) ng/L, (6.71 +/- 2.28) microg/L, respectively] were significantly higher than those of the controls [(0.128 +/- 0.029) ng/L, (4.31 +/- 1.13) microg/L respectively, P<0.05, P<0.01]. CONCLUSION: The increased levels of serum TNF-alpha, EGF in patients with DMLT may be related to the following causes: (1) Trichloroethylene and its metabolite may irritate the macrophagocyte and monocyte in human body to release TNF-alpha into blood stream; (2) Severe damage of epithelial tissue in patients with DMLT may promote more EGF synthesis to accelerate the regeneration and repair of epithelial tissue. PMID- 14761358 TI - [Detection of DNA damage induced by carbon disulfide in mice sperm with single cell gel electrophoresis assay]. AB - OBJECTIVE: To study the genotoxicity of carbon disulfide by detecting DNA damage in mice sperm with single-cell gel electrophoresis assay (SCGE). METHODS: SCGE was used to detect sperm DNA damage. The index of DNA damage, tail length and tail moment were used to evaluate the extent of DNA damage. RESULTS: In three dosage groups, the rate of DNA damage (67.14%, 84.29% and 91.00%, respectively), index of DNA damage intensity (507, 656 and 745, respectively), tail length (5.87, 8.81 and 13.49 microm, respectively) and tail moment (1.30, 1.63, 2.66 microm, respectively) were significantly increased, while the percentage of head of the comet was significantly decreased (84.55%, 73.84% and 55.71%, respectively). A significant changes were clearly observed in all dosage groups compared to those of the control group (P<0.05). CONCLUSION: SCGE which is a quick and sensitive method to detect DNA damage induced by CS2 may be used to monitor carcinogen and mutagen. PMID- 14761388 TI - [Damage and its protection from Electromagnetic Radiation]. PMID- 14761389 TI - [Effects of electromagnetic pulse on contents of amino acids in hippocampus of rats]. AB - OBJECTIVE: To investigate the relationship between the changes of amino acids contents in hippocampus of rats and electromagnetic pulse (EMP) exposure. METHODS: Rats were decapitated and hippocampus were removed after EMP (6 x 10(4) V/m, rise time 20 ns, pulse width 30 micro s, 5 pulses in 2 minutes) irradiation, and contents of amino acids were detected with high performance liquid chromatograpy (HPLC). RESULTS: The contents of aspartic acid (Asp) and glutamic acid (Glu) increased significantly 0, 3, 6 h after irradiation. The peak values of Asp [(17.25 +/- 1.63) pmol/ micro l] and Glu [(13.67 +/- 0.95) pmol/ micro l] were higher than those of control [(10.56 +/- 1.50), (6.94 +/- 1.10) pmol/ micro l respectively, P < 0.05]. Then both decreased gradually and reached the normal level 24 - 48 h after irradiation. The contents of glycine (Gly), taurine (Tau) and gamma-aminobutyric acid (GABA) also rose after exposure, the peak value of them [(4.51 +/- 0.60), (29.85 +/- 2.70), (5.14 +/- 0.73) pmol/ micro l respectively] were higher than those of control group [(2.18 +/- 0.31), (9.88 +/- 1.47), (2.84 +/- 0.67) pmol/ micro l, P < 0.05], then recovered 48 h after irradiation. The value of Glu/GABA increased immediately after exposure (3.45 +/- 0.25, P < 0.05), then decreased 24 h (1.62 +/- 0.23, P < 0.05) and recovered 48 h after exposure. CONCLUSION: The toxic effect of excess excitatory amino acids may be partly responsible for the early retardation (within 24 h) of learning of rats. PMID- 14761390 TI - [Effects of low frequency pulsed magnetic field on the proliferation and differentiation of HepG2 cells]. AB - OBJECTIVE: To study the effects of low frequency pulsed magnetic field on the proliferation and differentiation of HepG2 cells. METHODS: 3-(4,5-dimethylthiazol 2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) colorimetry method and ELISA assay of alpha-fetoprotein (AFP) were used to determine the cell proliferation and differentiation after the cells were exposed to pulsed magnetic fields with different frequency but the same field intensity. RESULTS: There were no significant differences in cell proliferation between sham and treated groups exposed to the field of 80 Hz, 1.55 mT for 1, 4, 8, 12, 24 h (P > 0.05). There were also no significant differences in cell proliferation and AFP secretion between sham and treated groups exposed to 16 Hz, 1.55 mT pulsed magnetic fields for 1, 4, 8, 24 h (P > 0.05). CONCLUSION: There were no "window effects" found in HepG2 cells proliferation or AFP secretion at 16 Hz and 80 Hz pulsed magnetic fields. PMID- 14761391 TI - [Effect of electromagnetic pulse irradiation on mice reproduction]. AB - OBJECTIVE: To evaluate the effect of electromagnetic pulse (EMP) irradiation on mice reproduction. METHODS: Female/male Kunming mice, 6 - 8 weeks old, prior to mating, or female after pregnancy were treated with whole body irradiation by 6 x 10(4) V/m electromagnetic pulse (EMP) for five times. The pregnant mice were killed on the 18th days, and teratological markers were analysed. RESULTS: EMP irradiation caused no significant changes in most of female organ weight and organ/body weight ratio. But it caused significant shortening in tail length of live foetus in the female mice before conception (prior to mating) or after pregnancy (P < 0.05), and obvious decrease in male offspring ratio (0.85 +/- 0.09 vs 1.09 +/- 0.17, P < 0.05). The male offspring ratio also significantly decreased (0.76 +/- 0.18 vs 1.09 +/- 0.17, P < 0.01) after male mice irradiated by EMP. The tail length of live foetus was shortened and male offspring sex ratio was increased after both male and female mice were irradiated by EMP. EMP irradiation also caused a significantly higher fetal death rate than normal control (P < 0.05). The embryo absorption rate was increased after irradiation except that was decreased in male mice. CONCLUSION: EMP irradiation has effect on pregnancy and offspring development in both male and female mice before mating and in female mice after pregnancy. PMID- 14761392 TI - [Effect of extremely low frequency magnetic fields on intracellular free calcium in HepG2 cells]. AB - OBJECTIVE: To study the effect of extremely low frequency magnetic fields on intracellular calcium concentration ([Ca(2+)]i). METHODS: Fura-2 loaded HepG2 cells were exposed to 1.55 mT (average value), 16 Hz pulsed magnetic fields for 60 min and to 300 mT, 2 Hz rotating magnetic fields for 5 min, and then [Ca(2+)]i was measured by fluorescence spectrophotometer. [Ca(2+)]i of HepG2 cells was also measured when they were exposed to 0.9 mT [root mean square (rms)], 16 Hz sinusoidal magnetic fields in real time. RESULTS: The R values (F(340) nm/F(380) nm) of the control and the exposed group were 2.4519 +/- 0.2378 and 2.5266 +/- 0.2915 respectively after HepG2 cells were exposed to 1.55 mT, 16 Hz magnetic fields, 1.365 0 +/- 0.0626 and 1.3602 +/- 0.0771 respectively to 300 mT, 2 Hz rotating magnetic fields. The ratios of the trendline slope [r((501 - 1,000)) / r((0 - 500))] from the data of R values were 1.1213 +/- 0.4559 and 1.0727 +/- 0.1971 respectively (P > 0.05), and the ratios of the intercept [b((501 - 1,000)) / b((0 - 500))] from the trendline were 0.9912 +/- 0.0098 and 0.9979 +/- 0.0060 (P > 0.05) when HepG2 cells were exposed to the 0.9 mT, 16 Hz sinusoidal magnetic fields. CONCLUSION: The effect of extremely low frequency magnetic fields on [Ca(2+)]i of HepG2 cells under the experimental condition has not been found. PMID- 14761393 TI - [Effects of extremely low frequency electromagnetic fields on the level of c-fos mRNA in brain and liver of mouse]. AB - OBJECTIVE: To study the effects of extremely low frequency electromagnetic fields (ELF EMFs) on c-fos gene expression in mouse brain and liver tissues. METHODS: Mice were exposed to 50 Hz sinusoidal 0.2 mT or 6.0 mT electromagnetic field for 2 weeks or 4 weeks. Competitive RT-PCR method was used to measure c-fos mRNA level. RESULTS: After exposure to 0.2 mT or 6.0 mT field for 2 weeks, c-fos mRNA levels in brain tissue [(0.0178 +/- 0.0076) amol/120 ng cDNA and (0.0092 +/- 0.0042) amol/120 ng cDNA respectively] were higher than that of control level [(0.0012 +/- 0.0005) amol/120 ng cDNA] (P < 0.05). In liver tissue c-fos mRNA levels [(0.0117 +/- 0.0055) amol/120 ng cDNA and (0.0148 +/- 0.0162) amol/120 ng cDNA respectively] were also higher than that of control level [(0.0005 +/- 0.0005) amol/120 ng cDNA] (P < 0.05). After exposure to 0.2 mT or 6.0 mT field for 4 weeks, c-fos mRNA levels in brain tissue [(0.0100 +/- 0.0054) amol/120 ng cDNA and (0.0198 +/- 0.0079) amol/120 ng cDNA respectively] were higher than that of control level [(0.0015 +/- 0.0008) amol/120 ng cDNA] (P < 0.05). In liver tissue the exposure induced much higher expression level [(0.0173 +/- 0.0122) amol/120 ng cDNA and (0.0133 +/- 0.0090) amol/120 ng cDNA respectively] while no expression was found in the control. CONCLUSION: Exposure to 50 Hz electromagnetic fields may induce up-regulation of c-fos transcription in mouse brain and liver tissue. PMID- 14761394 TI - [Effects of extremely low frequency electromagnetic fields on apoptosis and cell cycle of mouse brain and liver cells]. AB - OBJECTIVE: To study the effects of extremely low frequency electromagnetic fields (ELF EMFs) on apoptosis and cell cycle of mouse brain and liver cells. METHODS: Mice were exposed to 50 Hz, 0.2 mT or 6.0 mT electromagnetic fields for 2 weeks. TUNEL and flow cytometric methods were used to analyze apoptosis and cell cycle of brain and liver cells. RESULTS: After exposure to 0.2 mT and 6.0 mT ELF EMFs for 2 weeks, apoptosis rates of brain cells [(5.60 +/- 1.47)% and (4.73 +/- 0.48)% respectively] were higher than that of control [(2.90 +/- 0.75)%], and apoptosis rates of liver cells [(4.19 +/- 2.08)% and (3.38 +/- 0.65)% respectively] were higher than that of control [(1.84 +/- 0.76)%]. G0/G1 cell percentage of brain cells [(80.21 +/- 1.68)% and (79.54 +/- 0.56)% respectively] were higher than that of control [(76.85 +/- 0.83)%], and those of liver cells [(79.42 +/- 1.80)% and (80.47 +/- 1.79)% respectively] were higher than that of control [(73.36 +/- 3.10)%]. The above differences were all statistically significant as P < 0.05. At the same time S and G2 + M cell percentage of brain and liver cells were significantly decreased. CONCLUSION: Exposure to 50 Hz EMFs may alter cell cycle and induce apoptosis of mouse brain and liver cells. PMID- 14761395 TI - [Effects of extremely low frequency electromagnetic fields on male reproduction in mice]. AB - OBJECTIVE: To investigate the effects of extremely low frequency electromagnetic fields (ELF EMFs) on male reproduction in mice. METHODS: 94 adult male mice were exposed to 50 Hz sinusoidal electromagnetic fields of 0.2, 3.2 or 6.4 mT for 2 weeks or 4 weeks. Testicular histology and weight, sperm amount, sperm motility and morphology were measured. The percentages of different ploidy cells and cell phases, and DNA content of testis cells were estimated by flow cytometry. The micronucleus rate of bone-marrow cell was also observed. RESULTS: The testicular weight of the mice exposed to 6.4 mT for 4 weeks [(76.06 +/- 32.25) mg] was significantly lower than that of the control [(111.44 +/- 19.99) mg, P < 0.05]; no significant histopathological changes were observed on the testis in EMFs exposed mice;the sperm amount was decreased after EMFs exposure for 4 weeks, and those of the mice exposed to 0.2 mT and 6.4 mT for 4 weeks [(4.87 +/- 0.94) x 10(6)/ml and (4.30 +/- 1.89) x 10(6)/ml respectively] were significantly lower than that of the control [(6.67 +/- 0.70) x 10(6)/ml, P < 0.05]; the rates of sperm motility also showed a decline. After 0.2, 3.2 or 6.4 mT EMFs exposure for 2 weeks, the deformity rates of sperm [(7.416 +/- 3.352)%, (6.862 +/- 2.947)% and (8.112 +/- 4.615)% respectively] were significantly higher than that of the control [(4.098 +/- 2.028)%, P < 0.01]. Similarly, those of the mice exposed for 4 weeks [(10.267 +/- 3.836)%, (11.027 +/- 7.059)%, (8.814 +/- 3.678)% respectively] were higher than that of the control [(3.714 +/- 1.830)%]. After 6.4 mT exposure for 2 weeks, the percentages of 1C testis cells [(69.56 +/- 4.07)%] was significantly lower than that of the control [(73.45 +/- 3.10)%, P < 0.05]. There were not any remarkable changes in those of 2C, 4C cells. DNA content in different ploidy cells of the mice exposed to 6.4 mT was decreased. Moreover, the cell percentage in S phase was increased significantly (P < 0.01). CONCLUSION: ELF EMFs exposure may have some adverse effects on reproduction in mice. PMID- 14761396 TI - [Effect of low-intensity microwave radiation on proliferation of cultured epithelial cells of rabbit lens]. AB - OBJECTIVE: To compare the effects of different doses of microwave on the proliferative activity and cell cycle of cultured epithelial cells of rabbit lens, and to investigate the limit tolerant of microwave exposure. METHODS: Cultured epithelial cells of rabbit lens were exposed to microwave radiation with frequency of 2,450 MHz and power density of 0.10, 0.25, 0.50, 1.00, 2.00 mW/cm(2) for 8 h in vitro. HE staining was used to observe the morphological changes of lens epithelial cells, the proliferative activity and cell cycle were measured by MTT assay and PI fluorescent staining. RESULTS: 8 h after radiation, 0.50, 1.00 and 2.00 mW/cm(2) microwave could decrease the proliferation of lens epithelial cells, make the cells disordered arrangement, shrinkage, detachment, and inhibit the synthesis of cell DNA. The percentage of G(0)/G(1) phase cells were 71.95% +/ 2.12%, 75.68% +/- 3.35% and 82.40% +/- 8.68% respectively, which were higher than that in control group (61.68% +/- 5.76%, P < 0.05 or P < 0.01). The percentage of S phase cells were 19.32% +/- 3.07%, 16.08% +/- 4.91% and 12.98% +/ 8.08% respectively, which were lower than that in control group (28.05% +/- 5.12%, P < 0.05 or P < 0.01). No obvious changes could be detected in 0.10, 0.25 mW/cm(2) microwave groups (P > 0.05). CONCLUSION: Microwave exceeding 0.50 mW/cm(2) may make injury to lens epithelial cells after 8 hour radiation, which may be related to the effect of microwave radiation on cell cycle. PMID- 14761397 TI - [Changes of pulmonary tissue area density and their mechanisms in rats with hypoxic pulmonary hypertension]. AB - OBJECTIVE: To investigate the changes of pulmonary tissue area density and their mechanisms in rats with hypoxic pulmonary hypertension (HPH). METHODS: 54 Wistar rats were divided into hypoxia 10 d (n = 12), 20 d (n = 12), 30 d (n = 12) groups and control group (n = 18). The rats in hypoxia groups were exposed to a simulated hypoxia environment at a height of 5 km above sea level to establish HPH models. The changes in pulmonary tissue area density and pathological morphology were determined by image analysis, optical microscope, electron microscope and histochemistry. RESULTS: After hypoxia, the pulmonary tissue area density markedly increased on 10 d (27.08% +/- 1.29%, P < 0.05), especially on 20 d (31.33% +/- 0.27%) and 30 d (31.10% +/- 1.95%) while that in control group was 22.78% +/- 1.17% (P < 0.01). The area density on 20 d was higher than that on 10 d (P < 0.05) but there wasn't significantly different between 10 d and 30 d (P = 0.057) after hypoxia. Pathological examination showed: edema and collapse of pulmonary tissue, swelling and degeneration of type II alveolus epithelial cells (PII); congestion, accumulation of polymorphonuclear neutrophils and platelets in capillaries; swelling and degeneration of endothelial cells, thickening of basement membranes. CONCLUSIONS: Hypoxia can induce increase in pulmonary tissue area density and decrease in aerial exchange area in alveoli. These changes may be related to the pulmonary collapse caused by the damage of PII and pulmonary surfactant system, structural remodeling of small pulmonary arteries, increase in blood cells and protein granules in alveolar cavity. PMID- 14761398 TI - [Measurement of sound pressure level at outlet of external auditory canal and eardrum]. AB - OBJECTIVE: To evaluate the sound pressure level at outlet of external auditory canal and eardrum, and to observe the range of maximum frequency of sound effect of outlet ear canal, so as to provide exact data for designing personal noise rating instrument and studying the effect of environmental noise on human body, and to provide basis for calibration of noise sound pressure level. METHOD: Sound pressure level at eardrum and outlet of external auditory canal in 30 young people (60 ears) were measured by Audio Scan RM500 at 50 dB SPL narrow band noise with different frequencies (0.25 - 8.00 kHz). RESULTS: At the same frequency, sound pressure levels at eardrum were less than (0.6 +/- 2.6) dB SPL, at outlet of outlet ear canal less than (0.2 +/- 1.8) dB SPL, there were no significant differences between both sound pressure levels (P > 0.05). There were significant differences in sound pressure levels between both places at 1.50 - 8.00 kHz (P < 0.01). The maximum difference was (10.5 +/- 3.4) dB SPL at 2.00 kHz, and the minimum was (0.5 +/- 6.2) dB SPL at 0.50 kHz. CONCLUSION: The effects of increased sound pressure should be considered in the evaluation of environmental noise and the design for individual noise rating meter when the noise frequency was >or= 1.50 kHz. PMID- 14761399 TI - [Analysis of the sound-insulation effects of control rooms in workshops with noise]. AB - OBJECTIVE: To study the main factors affecting sound-insulation effects of control rooms in workshops with noise, so as to improve the protection. METHODS: The sound-insulation effects of 467 control rooms were determined, and different building materials, structures of door and window, airtight states etc. were analyzed. RESULTS: The affecting factors contributed to the sound-insulation effects (Eta(2)) were in the order: airtight states (0.168), building materials (0.080), structures of window and door (0.030, 0.029), sound pressure levels and frequency spectrum's characteristics (0.008, 0.006). Under airtight state, the sound-insulation effects of different building materials of the rooms were as follows: double bricks [(19.6 +/- 3.5) dB(A)]; single brick [(15.4 +/- 3.4) dB(A)]; plank [(13.1 +/- 1.6) dB(A)] or aluminum alloy plate with glass [(13.4 +/ 2.5) dB(A)] (P < 0.01). Of 4 group rooms, with the same structure of doors but double or single bricks of windows. 3 groups with dormant window had higher sound insulation effects [(15.9 +/- 2.8), (18.7 +/- 3.6), (19.3 +/- 2.5) dB(A)] than those with casement window [(14.1 +/- 2.4), (14.9 +/- 2.3), (16.5 +/- 2.4) dB(A)] (P < 0.01 or P < 0.05); 2 groups with dehydrated window [(18.7 +/- 3.3), (22.6 +/ 3.8) dB(A)] higher than those with dormant window [(15.9 +/- 2.8), (19.9 +/- 3.0) dB(A)] (P < 0.05). Of 6 group rooms, with the same structure of windows but double or single bricks of doors, only in 1 group with double-layer door had higher sound-insulation effect [(18.7 +/- 3.6) dB(A)] than that with single-layer door [(15.9 +/- 2.8) dB(A)] (P < 0.01). CONCLUSION: The control room should be designed rationally, kept airtight, according to the sound pressure levels and the condition of the workshop. PMID- 14761400 TI - [Co-detection of P21, P53 and HSP70 and their possible role in diagnosis of polycyclic aromatic hydrocarbons (PAHs)-related lung cancer]. AB - OBJECTIVE: To explore the biomarkers of early diagnosis in patients with polycyclic aromatic hydrocarbons (PAHs)-related lung cancer for the application to detection of occupational lung cancer or related lung cancer. METHODS: Western dot blotting was used to explore the expression of ras, p53 and heat stress protein 70 (HSP70) in 29 patients with PAHs-related lung cancer (LC), and 28 patients with non-cancerous pulmonary disease, and 30 healthy controls. RESULTS: The positive detection rates of P21, P53, and HSP70 in LC group (58.62%, 34.48%, 41.38% respectively) were higher than those in non-cancerous pulmonary disease group (14.29%, 7.14%, 10.71% respectively, P < 0.01). The sensitivity of P21, P53 and HSP70 were 58.62%, 34.48% and 41.38% respectively, negative predictive value (NPV) were 68.42%, 78.05% and 63.04% respectively. The co-detection of the three proteins mentioned above produced a sensitivity of 82.76% with a NPV of 78.26% (P < 0.05). Of 18 cases of LC with negative cytology, 13 (72.22%) were found HSP21, P53 or HSP70 positive. CONCLUSIONS: Co-detection of the P21, P53, and HSP70 may be used as the screening marker for diagnosis of PAHs-related lung cancer, and may supplement the diagnostic value of conventional cytology. PMID- 14761401 TI - [The expression of cytochrome P450 1A1 of endothelial cells induced by benzo(a)pyrene]. AB - OBJECTIVE: To explore the effect of benzo(a)pyrene (BaP) on the expression and the activities of cytochrome P450 1A1 (CYP1A1) of porcine aortic endothelial cells. METHODS: Porcine aortic endothelial cells were cultured in vitro, and treated with different concentrations of BaP (0, 0.5, 1.0, 5.0, 10.0 micro mol/L) for 24 hours, CYP1A1 expression was determined by Western blot and immunohistochemistry. At the same time, the ethoxyresorufin-o-deethylase (EROD) activities were measured by spectrofluorometer. RESULTS: By Western blot, the expression of CYP1A1 of control cells was not found, but the expression of CYP1A1 of cells treated with BaP was found; By immunohistochemistry, only part of endothelial cells treated with BaP had positive expression of CYP1A1. The peak activities of EROD induced by BaP was at the concentration of 0.5 - 1.0 micro mol/L. CONCLUSION: BaP could induce part of endothelial cells to synthesize CYP1A1. BaP of 0.5 - 1.0 micro mol/L could induce peak activities of EROD. PMID- 14761402 TI - [Spectral analysis of electromyography of low back muscle fatigue induced by simulated driving]. AB - OBJECTIVE: To evaluate the effects of different vibration frequency on the back muscle fatigue during simulated driving. METHODS: Thirty-six subjects performed three simulated driving experiments under three vertical vibration frequencies which were 1.8, 4.0, 6.0 Hz respectively and the driving time was 90 minutes. At the same time the electromyography of low back was recorded. RESULTS: The median frequencies calculated from the power spectrum were decreased exponentially under three vertical vibration frequencies, especially under 4.0 Hz vibration frequency. CONCLUSION: The 4.0 Hz vibration frequency has the most important effect on the back muscle fatigue under simulated driving condition. PMID- 14761411 TI - [Molecular basis of UVA-induced skin aging and arotinoid ethyl ester protection]. PMID- 14761423 TI - [The inhibitory effect of Lu-Duo-Wei on carbon disulfide-induced generation of hydroxyl radicals]. AB - OBJECTIVE: To study the scavenging effects of Lu-Duo-Wei, thiourea, superoxide dismutase, and sodium azide on carbon disulfide-induced generation of hydroxyl radicals. METHODS: Phenanthroline-CuSO(4)-Vit C-H(2)O(2) chemiluminescence system (PHEN system) containing alcohol was established to probe the influence of various concentrations of carbon disulfide on hydroxyl radicals emission intensity and the scavenging effects of Lu-Duo-Wei and other antioxidants on carbon disulfide-induced hydroxyl radicals were observed. RESULTS: The average emission intensity of PHEN system containing alcohol appeared lower luminescence [91.03 x 10(3) (cp6s)] and longer time (75 s) to get the peak than the system without alcohol [96.11 x 10(3) (cp6s), 55 s]. The specific scavenger of hydroxyl radical, thiourea, showed clear inhibitory effect on the system. Carbon disulfide in the range of 40 - 160 mmol/L promoted the generation of hydroxyl radical, however, this effect could be efficiently inhibited by thiourea. 160 mmol/L carbon disulfide in PHEN system without copper seemed as an activator to promote the luminescence, while in PHEN system withdrawing phenanthroline appeared some weak action of luminescence agent at low concentration. Meanwhile, Lu-Duo-Wei may efficiently scavenge hydroxyl radicals induced by carbon disulfide in PHEN system but superoxide dismutase and sodium azide had little effects on the system. CONCLUSION: Carbon disulfide may induce PHEN system to generate hydroxyl radicals and Lu-Duo-Wei may efficiently scavenge these free radicals and play an important role in protection against oxidative injury induced by carbon disulfide. PMID- 14761424 TI - [The treatment with allogeneic peripheral blood stem cell transplantation for benzene-induced severe aplastic anemia]. AB - OBJECTIVE: To evaluate the efficacy of treatment with allogeneic peripheral blood stem cell transplantation for benzene-induced severe aplastic anemia. METHODS: HLA-compatible sibling (pregnancy) allogeneic peripheral blood stem cell transplantation (Allo-PBSCT) was successfully performed for a patient with severe aplastic anemia caused by benzene poisoning. 9.41 x 10(8) nucleated cells/kg, 12.49 x 10(6) CD(34) positive cells/kg and CFU-GM 8.2 x 10(5)/kg were infused. The patient was treated with cyclophosphamide (120 mg/kg), total body radiation (8 Gy) and anti-lymphocyte globulin (60 mg/kg) before transplantation. Donor buffy coat cells (9.02 x 10(8) nucleated cells/kg, 10.62 x 10(6) CD(34) positive cells/kg, 6.3 x 10(5) CFU-GM/kg) were infused again on day 18 after transplantation to prevent from graft failure. Graft versus host disease prophylaxis consisted of both methotrexate and cyclosporin A. RESULTS: The lowest ANC was 0, the lowest platelet was 3 x 10(9)/L after transplantation. The patient achieved an ANC of greater than 0.5 x 10(9)/L from 21st day, and the platelet of greater than 50 x 10(9)/L from 28th day after transplantation. Grade I cGVHD was found the fourth month after grafting. Examination of recipient's bone marrow cells showed a normal 46, XX (presumably marrow donor) karyotype. Blood group changed from B to O. CONCLUSION: This is the first case reported in China showing a successful treatment of benzene-induced severe aplastic anemia with allo-PBSCT. Allo-PBSCT may be an effective remedy for this kind of patients. PMID- 14761426 TI - [Effect of immunocyte therapy on benzene-induced bone marrow haemopoietic dysfunction]. AB - OBJECTIVE: To explore the effect of treatment with immunocyte therapy on benzene induced haemopoietic dysfunction. METHODS: Mono-nuclear cells (MNC) were separated from 40 - 50 ml peripheral blood in patients and mixed with interleukin 2 and granulocyte macrophage colony stimulating factor (GM-CSF) for six day cultivation. The new formed immunocytes were collected and transfused into the patients. Bone marrow aspiration and biopsy were taken before and after therapy for all patients with severe benzene poisoning. Blood samples were stained by flow cytometry for detecting CD(4) and CD(8) positive cells. RESULTS: Of 20 patients with chronic benzene poisoning, 9 were severe benzene poisoning. All examination including blood count, bone marrow biopsy and T cell subpopulation restored to normal after immunocyte therapy. Laboratory tests (liver and kidney function, and myocardial enzymes) were observed periodically and showed normal during therapy. Follow-up study (the longest time was more than 15 months) showed that bone marrow haemopietic function of all treated patients were in normal range. CONCLUSION: Bone marrow haemopoietic dysfunction caused by benzene poisoning may be closely related to disorder of immune function. Immunocyte therapy may significantly improve bone marrow haemopoietic dysfunction induced by benzene poisoning. PMID- 14761430 TI - [Effects of fluoride on proliferation and differentiation of rat osteoblasts in vitro and the antagonistic action of vitamin C]. AB - OBJECTIVE: To study the effects of fluoride on the proliferation and differentiation of osteoblasts in sucking rats and the antagonism of vitamin Cin vitro. METHODS: The enzyme digesting method was used to isolate the rat osteoblasts; the proliferative response was determined by the percents of reduced alamarBlue; the activity of alkaline phosphatase (ALP) was measured by ELISA method. RESULTS: The proliferation of sucking rat osteoblasts was increased at 0.10 - 1.00 mmol/L of NaF, whereas inhibited at >or= 2.00 mmol/L. ALP activity was increased at 0.01 - 0.05 mmol/L of NaF, and decreased at >or= 0.10 mmol/L. The inhibition on proliferation and differentiation at 2 mmol/L NaF was antagonized by vitamin C. CONCLUSION: Fluoride had a two-phase effect on osteoblasts, vitamin C could antagonize the inhibitory effect of higher concentration of fluoride on proliferation and differentiation of osteoblasts. PMID- 14761428 TI - [Detection of myocardial lesion in patients with acute ammonia poisoning]. AB - OBJECTIVE: To find a better index for detecting myocardial lesion in patients with acute ammonia poisoning. METHODS: Cardiac troponin T (cTnT), creatine kinase (CK), creatine kinase-MB isoenzyme (CK-MB), and electrocardiogram (ECG) were determined and compared in patients with acute ammonia poisoning and healthy controls. RESULTS: In severe ammonia poisoning patients, the levels of cTnT [(1.285 +/- 2.650) micro g/L], CK [(257.636 +/- 362.719) IU/L], CK-MB [(20.909 +/ 19.770) IU/L] were significantly higher than those in healthy control [(0.035 +/ 0.014) micro g/L, (82.050 +/- 36.302) IU/L, (8.350 +/- 2.455) IU/L respectively, P < 0.05]. The levels of cTnT, CK and CK-MB in mild ammonia poisoning patients [(0.039 +/- 0.016) micro g/L, (78.200 +/- 28.401) IU/L and (8.467 +/- 2.669) IU/L respectively], and moderate ammonia poisoning patients [(0.054 +/- 0.043) micro g/L, (99.786 +/- 71.941) IU/L and (9.429 +/- 3.857) IU/L were not significantly different from those of healthy controls (P > 0.05)]. cTnT positive detection rate (68.2%) was significantly higher than CK (36.4%), CK-MB (31.2%) and ECG (31.2%) (P < 0.05). CONCLUSION: cTnT is a better index to detect myocardial lesion in severe ammonia poisoning. PMID- 14761431 TI - [Publication of a sortable and searchable database on CD (Database of occupational medicine: regulation*standard*guideline)]. PMID- 14761432 TI - [Effect of differentiation inducer and heat stress on the expression of JWA protein and Hsp70 of K562 cells]. AB - OBJECTIVE: To study the expression of JWA protein and heat shock protein (Hsp70), and to explore these relationship and the possible mechanism of JWA gene involved in induced differentiation and heat stress (42 degrees C) of K562 cells. METHODS: The models of differentiation and heat stress of K562 cells were established. Western blot was used for detecting expressed proteins of JWA gene, Hsp70, heat shock factor (HSF1 and HSF2). RESULTS: (1) Under the condition of differentiations induced by TPA (100 ng/ml), hemin (3 x 10(-5) mol/L), Ara-C (80 ng/ml), adriamycin (4 x 10(-8) mol/L), ATRA (1 x 10(-6) mol/L) and As(2)O(3) (1 x 10(-6) mol/L) for 48 h respectively, the expression of JWA protein and Hsp70 were more significantly increased than control; the level of HSF2 protein was increased by inductions of hemin, Ara-C and adriamycin, respectively. (2) After heat exposure to 42 degrees C for 10, 20, 30, 45, 60, 90 min, and heat exposure to 39 degrees C, 42 degrees C, 45 degrees C, the trend of changing in expression of Hsp70 was similar to that of JWA protein, and HSF1 was expressed in earlier stage. CONCLUSION: The expression of JWA protein and Hsp70 were upregulated in induced differentiation and in heat stress, and the change of expression of JWA protein were similar to that of Hsp70, but the intracellular transduction signal pathways involved may be various. JWA might not be specifically related with both HSF1 and HSF2. PMID- 14761433 TI - [A study on the genital system injury induced by nickel sulfate in male rats]. AB - OBJECTIVE: To study the mechanism of the genital system damage by nickel sulfate in male rats in order to provide the laboratory theoretical evidence for the prevention and cure of nickel genital toxicity. METHODS: Three groups of rats were injected intraperitoneally with nickel sulfate at dose of 1.25, 2.50, 5.00 mg/kg respectively for two weeks. The content of testicle nickel and blood serum testosterone (T), follicle-stimulating hormone (FSH), luteinizing hormone (LH) were assessed with atomic absorption spectrum and radioimmuno-assay, meanwhile the activity of nitric oxide synthase (NOS) and the content of nitric oxide (NO) were measured by enzyme method. RESULTS: The contents of testicle nickel [(0.22 +/- 0.03), (0.34 +/- 0.04), (0.41 +/- 0.02) micro g/g respectively] were increased, but the content of T, TSH, LH in blood serum were reduced; the activities of NOS in testicle tissue [(33.65 +/- 2.93), (26.53 +/- 9.52), (10.20 +/- 2.74) U/g respectively] were inhibited by nickel sulfate and the contents of NO [(0.26 +/- 0.03), (0.18 +/- 0.05), (0.15 +/- 0.02) mmol/g respectively] were decreased (P < 0.01). CONCLUSION: Nickel-induced genital system injury of male rats may be related to the decrease in the contents of T, TSH, LH, and the inhibition on NOS, as well as the fall of NO content. PMID- 14761434 TI - [Investigation of benzene component induced indoor air pollution]. PMID- 14761435 TI - [The effect of overtraining on human sperm chromatin structure]. AB - OBJECTIVE: To identify the effects of overtraining on human sperm DNA. METHODS: Molecular epidemiological investigation of 249 men from different groups (training and non-training) was carried out by using flow cytometer to detect the integrity and damage of in situ DNA of sperm nucleus, and sperm chromatin structure assay was performed. RESULTS: The average COMPalpha(t) in training group was 11.02% while that in control group was 5.90% (P < 0.01). COMPalpha(t) was significantly correlated with sperm activity (r = 0.41, P < 0.05). CONCLUSION: Overtraining could induce sperm DNA injury and affect sperm activity, thus to decrease the potentiality of reproduction. PMID- 14761436 TI - [Effects of lead exposure on nitric oxide synthase activity in different brain regions of developmental rat]. AB - OBJECTIVE: To observe the influence of lead exposure on the activity of nitric oxide synthase (NOS) in different brain regions of rat. METHODS: By establishing a series of rat models exposed to different low levels of lead (drinking water containing 0.025%, 0.050%, 0.075% of lead acetate) during developing period, NOS activities in hippocampus, cerebellum, cerebral cortex and brain stem were studied. RESULTS: On the 21st day after birth, NOS activities in hippocampus of three levels of lead exposed groups [(1.53 +/- 0.20), (1.66 +/- 0.23), (1.88 +/- 0.32) U/mg pro respectively], and in cerebellum [(0.87 +/- 0.24), (0.85 +/- 0.09), (0.91 +/- 0.18) U/mg pro respectively] were significantly lower than those of control group [(2.36 +/- 0.18), (1.41 +/- 0.18) U/mg pro, respectively, P < 0.01]. NOS activities in cerebral cortex of 0.075% group [at 7, 14, 21 d of age [(1.29 +/- 0.14), (1.03 +/- 0.15), (0.69 +/- 0.10) U/mg pro] were significantly lower than those in control group [(2.54 +/- 0.31), (1.64 +/- 0.22), (1.24 +/- 0.14) U/mg pro respectively], and 0.025% group [(2.42 +/- 0.19), (1.59 +/- 0.17), (1.27 +/- 0.12) U/mg pro respectively], and 0.050% group [(2.56 +/- 0.53), (1.77 +/- 0.19), (1.24 +/- 0.10) U/mg pro respectively, P < 0.05]. There were no significant differences among control, 0.025%, and 0.050% groups (P > 0.05). Lead exposure had no influence on NOS activity in brain stem at the same age (P > 0.05). CONCLUSION: NOS activities in hippocampus, cerebellum and cerebral cortex were inhibited by low level lead exposure and the degree of the effect was related to Pb exposure time and/or level of Pb exposed. PMID- 14761438 TI - [Analysis of severe accidents in chemical factory from 1983-1999 in China]. PMID- 14761437 TI - [Effects of 2,450 MHz microwave on DNA damage induced by three chemical mutagens in vitro]. AB - OBJECTIVE: To study the combined damage-effects of low-intensity 2,450 MHz microwave (MW) with three chemical mutagens on human lymphocyte DNA. METHODS: DNA damage of lymphocytes exposed to microwave and(or) with chemical mutagens were observed at different incubation time (0 h or 21 h) with comet assay in vitro. Three combination-exposure ways of MW with chemicals were used: MW irradiation before chemical exposures, simultaneously exposed to MW and chemicals and MW irradiation after chemical exposures. The three chemical mutagens were mitomycin C (MMC, DNA crosslinker), bleomycin (BLM, radiometric agent), methyl methanesulfonate (MMS, alkylating agent). The exposure time of MW and chemical mutagens were 2 h and 3 h respectively. RESULTS: The differences of comet tail length between MW group and control group were not significant when lymphocytes were incubated for 0 h or 21 h (P > 0.05). However, when lymphocytes were incubated for 21 h with 30.00 micro mol/L of MMC, the comet tail lengths of MW + MMC group, MW-MMC group and MMC + MW group were (18.00 +/- 5.96), (21.79 +/- 11.47) and (22.32 +/- 8.10) micro m respectively; while with 3.00 micro mol/L of MMC, the comet tail lengths were (8.99 +/- 3.75), (12.40 +/- 5.35) and (14.00 +/- 5.38) micro m respectively, which were significantly higher than those of corresponding MMC groups [(9.42 +/- 3.34) and (6.50 +/- 2.89) micro m, P < 0.01 or P < 0.05]. The DNA damage of MW plus BLM groups and MW plus MMS groups were not significantly different from the corresponding BLM and MMS groups (P < 0.05). CONCLUSION: 2 450 MHz MW (5 mW/cm(2)) did not induce DNA damage directly, but could enhance the DNA damage effects induced by MMC. The synergistic effects of 2 450 MHz MW with BLM and MMS were not obvious. PMID- 14761439 TI - [Investigation of payments for disabled workers in Ben-Xi metal foundry from 1992 2000 ]. PMID- 14761440 TI - [Investigation of air pollution poisoning and air quality in factories in Zhu hai]. PMID- 14761441 TI - [Observation of lymphocyte ultrastructure in x-ray diagnostic technicians]. PMID- 14761442 TI - [Study of correlation between vitiligo and the rubber industry]. PMID- 14761443 TI - [Gene mutation and sister chromatid exchange induced by propylene oxide]. PMID- 14761444 TI - [Study of muscle strength and spinal curvature in young soldiers with back pain]. PMID- 14761445 TI - [Acrylonitrile induced poisoning effects on male reproductive system from occupational exposure]. PMID- 14761446 TI - [Comparison of amount of protein in urine before and after centrifuging]. PMID- 14761447 TI - [Effect of chlorine gas pulmonary emphysema on IL-8 and TNF in rats]. PMID- 14761448 TI - [Study of the use of 3-d targeted radiotherapy to control silicosis related local lung cancer]. PMID- 14761449 TI - [Application of hemofiltration in acute poisoning therapy]. PMID- 14761451 TI - [8 cases of carbon monoxide induced cardiac infarction]. PMID- 14761450 TI - [Clinical study of 22 cases of acute ammonia poisoning]. PMID- 14761452 TI - [One case of drug poisoning]. PMID- 14761453 TI - [Successful treatment in 46 cases of acute poisoning ]. PMID- 14761454 TI - [Study of CK,AST levels and ECG in 18 cases of acute poisoning ]. PMID- 14761455 TI - [Mushroom acute poisoning in 5 cases ]. PMID- 14761456 TI - [Evaluation of safety features after renovation of an X-ray facility]. PMID- 14761457 TI - [Comparison of breathing and different methods of dust collection]. PMID- 14761458 TI - [Aluminum detection in urine by direct absorption spectroscopy ]. PMID- 14761459 TI - [Amitraz detection in workplace air by gas chromatography]. PMID- 14761460 TI - [Detection of lead in workplace air using hydroxyl reaction by atomic fluorescent spectrum]. PMID- 14761461 TI - [Gas chromatographic detection of dicyclopentadiene in workplace air]. PMID- 14761462 TI - [Study of gel electrophoresis storage time for a single cell using alcohol desiccation]. PMID- 14761463 TI - [Detection method for N,N-DMF in the air by gas chromatography in microtubule]. PMID- 14761464 TI - [Sulphuric acid detection in the workplace by atomic fluorescent spectrum ]. PMID- 14761465 TI - [Trends of International coordination in risk evaluation and management of chemical compounds]. PMID- 14761466 TI - [Status of prevention and treatment of pesticide poisoning in the general public]. PMID- 14761467 TI - [Tetramine poisoning clinical characteristics and current treatment in China]. PMID- 14761468 TI - [Evaluation of risks of volatile organic chemicals as detergent substitute in industry]. PMID- 14761469 TI - [Current status of industrial chemical exposure level management]. PMID- 14761470 TI - [Strategy considerations in public health in China]. PMID- 14761471 TI - [Occupational health of medical personnel]. PMID- 14761472 TI - [Epidemiological study of gem worker's silicosis]. AB - OBJECTIVE: To study the hazards of gem dusts and the epidemiological features of the occurrence of gem worker's silicosis (GWS). METHODS: Epidemiological investigation in a gem factory in Guangdong province was carried out. RESULTS: The average dusts concentration in workshop was 2.3 mg/m(3), average free silica content was 94.64%. There were 47 patients diagnosed as silicosis. The prevalence of GWS was 4.58% (47/ 1,027). The age of patients suffering from silicosis and the duration of dust exposure were (27.04 +/- 3.25) years and (4.67 +/- 1.17) years respectively. Chest radiographic examination showed mainly small round opacities "q". The incidences of GWS complicated by pulmonary tuberculosis, spontaneous pneumothorax and pulmonary emphysema were 10.64%, 10.64%, 44.68% respectively. CONCLUSION: The hazards of gem dust to workers might be serious. Higher incidence, younger age suffered from silicosis, shorter duration of dust exposure, more complications, and poorer prognosis in gem workers appeared to be the main epidemiological features of GWS in Guangdong. PMID- 14761473 TI - [Case of occupational autoimmune alveolar pneumonia]. PMID- 14761474 TI - [Study on the expression of C-erbB-2 gene in coal miners with pneumoconiosis complicated by pulmonary cancer]. AB - OBJECTIVE: To study the difference in the expressions of C-erbB-2 gene between the coal miners with pneumoconiosis complicated by pulmonary cancer and the controls with single pulmonary cancer, and its relation to clinical pathology. METHOD: Measuring the expressions of C-erbB-2 in 32 cases of pneumoconiosis complicated by pulmonary cancer and those in 30 cases of pulmonary cancer by means of immunohistochemistry. RESULTS: The positive expression rate in 32 cases of pneumoconiosis complicated by pulmonary cancer was 53.13% whereas that in 30 cases of single pulmonary cancer was 26.67% (P < 0.05); the positive expressions of C-erbB-2 in patients with lymph node metastasis (70.59% in pneumoconiosis group, 50.00% in controls) were significantly different from those without lymph node metastasis (33.33% in pneumoconiosis group, 11.11% in controls) (P < 0.05). The prognosis on patients with positive expressions of C-erbB-2 was poor, and was not related to pathologic category. CONCLUSION: C-erbB-2 gene may be an important regulating gene in the coal miners with pneumoconiosis complicated by pulmonary cancer, and as a reference index to determine lymph node metastasis and prognosis. PMID- 14761475 TI - [31 cases of pneumoconiosis complicated with acute myocardial infarction in coal miners]. PMID- 14761476 TI - [Study on the pulmonary fibrogenetic effect induced by rush-mat dust in rats]. AB - OBJECTIVE: To investigate the fibrogenetic effects induced by rush-mat dust in rats. METHODS: SD rats were treated with 50 mg of rush-mat dust per rat by intra tracheal instillation, sacrificed 3, 6, and 12 months respectively after exposure. The lung tissue and lung lymph-node were taken out for pathological and electron microscopic examination. The content of collagen and ceruloplasmin (CP) in lung tissues were also determined. RESULTS: After treatment for 12 months, fresh wet lung weight in rush-mat dust group [(2.69 +/- 0.22) g] was higher than those in saline group [(1.87 +/- 0.25) g], TiO(2) group [(2.25 +/- 0.26) g], but lower than that in SiO(2) group [(11.41 +/- 1.63) g]; dry lung weight in rush-mat dust group [(0.47 +/- 0.03) g] was higher than those in saline group [(0.32 +/- 0.03) g], TiO(2) group [(0.41 +/- 0.08) g], but lower than that in SiO(2) group [(2.06 +/- 0.28) g]; lung collagen content in rush-mat dust group [(103.08 +/- 14.79) mg] was higher than those in saline group [(75.96 +/- 13.91) mg, TiO(2) group [(85.84 +/- 17.62) mg], but lower than that in SiO(2) group [(497.50 +/- 100.80) mg]; CP content in rush-mat dust group [(18.03 +/- 1.87) U/L] was higher than those in saline group [(15.05 +/- 2.24) U/L], TiO(2) group [(16.92 +/- 1.67) U/L], but lower than that in SiO(2) group [(25.37 +/- 3.58) U/L], P < 0.05 or P < 0.01. Pathological examination showed lung macrophage alveolitis, broadening of alveolar interval, one to two grade of silicotic nodes and increased amount of type II epithelial cell in alveolar as well as slight collagenous fibrosis in lung tissue of rush-mat dust group. Under electron microscope, primary and secondary lysosome and medullary sheath-like phagocytic residual body were found in lung tissue of rush-mat dust group, meanwhile the amount of type II alveolar epithelial cell and collagen fiber were slightly increased but these changes were less than those of quartz group. CONCLUSION: The rush-mat dusts have slight pulmonary fibrogenetic effect on rat. PMID- 14761477 TI - [Morphological transformation of mouse NIH3T3 cells induced by nickel refining dusts in vitro]. AB - OBJECTIVE: To study the biological effects of nickel-refining dust. METHODS: The cell phagocytosis, transformation activity, and cytotoxicity of the mouse NIH3T3 cells treated with nickel-refining dusts from two nickel-refining factories in China were observed, and the risk of carcinogenicity was studied. RESULTS: (1) Two samples of nickel-refining dusts could be phagocytosed by mouse NIH3T3 cells with different phagocytizing rates of 69.0% and 39.0% at 100.000 micro g/ml, and 78.0% and 47.0% at 200.000 micro g/ml respectively. The relative clone formation rates at 12.500 micro g/ml to 100.000 micro g/ml were 71.1% to 3.9% and 84.4% to 9.1%, respectively. The cytotoxicity expressed by clone formation rate was similar to that of Ni(2)O(3), but higher than that of TiO(2) and lower than the positive control of N-methyl-N'-nitro-N-nitroso-guanidine (MNNG). (2) MNNG, Ni(2)O(3) and the two samples of nickel-refining dusts could induce morphological transformation in NIH3T3 cells. The transformation rate at 12.500 micro g/ml to 50.000 micro g/ml were 1.9% to 3.6% and 0.9% to 2.5% respectively in a dose dependent manner. (3) The NIH3T3 cells treated by MNNG and nickel-refining dusts could induce Con A agglutination, and may form as clone in soft agar. This finding proved the reliability of the transformed clone. CONCLUSIONS: The present study for the first time demonstrate that nickel-refining dusts have cell transformation activity. The findings provide a new experimental evidence for the carcinogenic risk of nickel-refining dusts, and for the aetiology of lung cancer in nickel-refining workers. PMID- 14761478 TI - [The effect of silicon dioxide on the activation of nuclear factor-kappaB in THP 1 cells]. AB - OBJECTIVE: To study the effect of silicon dioxide (SiO(2)) on the activation of nuclear factor-kappaB (NF-kappaB) in THP-1 cell line. METHODS: THP-1 cells were incubated with a series of doses of SiO(2) (0, 100, 200 micro g/ml). The location of NF-kappaB p65 subunit (NF-kappaB/p65) in THP-1 cells was detected by immunofluorescence and laser scanning confocal microscope (LSCM). The expression of NF-kappaB/p65 in nuclei was measured by Western blot analysis. RESULTS: The majority of fluorescein isothiocyanate (FITC)-labelled NF-kappaB/p65 located in the nuclei 30 min after stimulation by 100 micro g/ml SiO(2), whereas the FITC labelled NF-kappaB/p65 were mainly seen in the plasma of normal control cells. The expression of NF-kappaB/p65 in THP-1 nuclear protein was low in control group (0 micro g/ml SiO(2)) while it increased after stimulation by 100 micro g/ml SiO(2) and 200 micro g/ml SiO(2) for 15 min and 30 min. The level of NF kappaB/p65 was comparatively increased with the increasing of doses and time. Lipopolysaccharides (LPS), an activator of NF-kappaB, had similar effect as SiO(2) on the activation of NF-kappaB/p65 in THP-1 cells. CONCLUSION: SiO(2) could activate and internalize NF-kappaB in the THP-1 cell line. PMID- 14761479 TI - [Immunohistochemical method for the detecting expression of transforming growth factor-beta1 in lung tissues of silica-treated mice]. AB - OBJECTIVE: To investigate the protein expression of transforming growth factor beta(1) (TGF-beta(1)) in lung tissues of silica-treated mice. METHODS: The experimental mice were divided into control and silica groups. 0.2 g/kg body weight of silica was injected intratracheally in mice of silica group. Samples of lung tissue were collected 1, 3, 5, 7, 14 and 28 d after injection. The immunohistochemical method was used to analyze the protein expression of TGF beta(1). RESULTS: In control mice, the expression of TGF-beta(1) in lung tissue was slightly positive while it was markedly increased in silica-treated mice. The expression was significantly elevated from the 7th day to 14th day. The expression in alveolar macrophages reached the peak on the 5th day [(93.4% +/- 2.8%) vs (42.2% +/- 12.0%), P < 0.01]. CONCLUSION: TGF-beta(1) may play an important role in early development of silicosis. PMID- 14761480 TI - [RT-PCR method for detecting the expression of transforming growth factor-beta1 gene in lung tissues of silica-treated mice]. AB - OBJECTIVE: To investigate the gene expression of transforming growth factor beta(1) (TGF-beta(1)) in lung tissues of silica-treated mice. METHODS: The experimental mice were divided into control and silica group. 0.2 g/kg body weight of silica was injected intratracheally in silica group. Samples of lung tissue were collected 1, 3, 5, 7, 14 and 28 d after injection. RT-PCR method was used to analyze the gene expression of TGF-beta(1) in lung tissue of silica treated mice. RESULTS: The expression of TGF-beta(1) gene in lung tissue elevated from the 3rd day (1.20 +/- 0.15) and the peak value was on the 7th day (1.74 +/- 0.19). Then the expression decreased from the 14th to 28th day. But there was still higher than control until the 28th day. CONCLUSION: TGF-beta(1) may play an important role in silica-induced pulmonary fibrosis. PMID- 14761481 TI - [Delta-aminolevulinic acid urine level in people with no lead contact in China ]. PMID- 14761482 TI - [Effect of paraoxon and chlorpyrifos on the nicotinic autoreceptor function in rat cortical synaptosomes]. AB - OBJECTIVE: To explore the effect of organophosphorus insecticides (OPs) on the nicotinic autoreceptor function (NAF) in rat cortical synaptosomes and to understand alternative target of the OPs for human and other animals. METHODS: In vitro experiment, synaptosomes from the rats were incubated with [(3)H] choline and then superfused with physiological buffer. The [(3)H] acetylcholine release from the synaptosomes after the addition of paraoxon or chlorpyrifos to the superfusion system was recorded and the changes of NAF were calculated. In vivo experiment, NAF and acetylcholinesterase (AChE) activity in the cortical synaptosomes in the adult rats dosed with chlorpyrifos were also determined 96 h after OPs treatment. RESULTS: Paraoxon caused slight effect on NAF (average inhibition rate: < 23%) while chlorpyrifos oxon caused > 100% of inhibition on NAF in vitro. Chlorpyrifos markedly reduced NAF by 66% 96 h after treatment and inhibited the AChE activity by 91% in vivo. CONCLUSION: The OPs may have different effects on the NAF of rat cortical synaptosomes while chlorpyrifos may certainly inhibit the NAF in cortical synaptosomes of adult rats. PMID- 14761483 TI - [Case of CO poisoning of cement factory workers ]. PMID- 14761484 TI - [Potential biochemical mechanisms of neuromuscular junction transmission dysfunction induced by organophosphorus insecticides]. AB - OBJECTIVE: To explore the potential biochemical mechanisms of neuromuscular junction transmission (NMJT) dysfunction induced by organophosphorus insecticides. METHODS: Ten rats were dosed with phoxim (1,144 mg/kg) and 5 of them developed myasthenia. The NMJT function was evaluated by the mean consecutive differences (MCD) measured by stimulation single fiber electromyography (SSFEMG) with the frequency of stimuli at 20 Hz. The activity of Ca(2+)-ATPase, Na(+), K(+)-ATPase, cAMP dependent protein kinase (PKA), Ca(2+)/phospholipid dependent protein kinase (PKC), and tyrosine protein kinase (TPK) were determined. RESULTS: In comparison with the control and non-myasthenic rats, the results in myasthenic rats showed that: (1) the MCDs increased; (2) the activities of Ca(2+)-ATPase, Na(+), K(+)-ATPase and PKA decreased and were negatively correlated with MCD; (3) the activities of PKC and TPK increased, and were positively correlated with MCD; (4) the phosphorylation of serine residuals in sarcolemma was weaker and the phosphorylation of tyrosine residuals was stronger. CONCLUSIONS: The NMJT dysfunction is likely associated with the decrease in Ca(2+)-ATPase and Na(+), K(+)-ATPase activity. The acceleration of desensitization and prolongation of resensitization of nicotinic acetylcholine receptors occur following the increase in tyrosine phosphorylation and the decrease in serine phosphorylation induced by OPs poisoning. PMID- 14761485 TI - [Effects of deltamethrin on cell survival rate and intracellular free Ca2+ concentration in primary cultured astrocytes of rat]. AB - OBJECTIVE: To study the effects of deltamethrin (DM) on cell survival rate and intracellular Ca(2+) ([Ca(2+)]i) concentration in primary cultured astrocytes of rat. METHODS: The cell survival rate was measured by Typan Blue assay; the intracellular [Ca(2+)]i concentration was determined by the fluorescent Ca(2+) indicator Fura-2/AM. RESULTS: The survival rate of astrocytes was decreased to 91.9% after astrocytes were incubated with 1 x 10(-5) mol/L DM for 72 h (P < 0.05). The cell survival rates were 89.0%, 84.8%, 81.2% and 79.2% respectively when astrocytes were administered with 1 x 10(-4) mol/L DM for 4, 12, 24 and 72 h, which were remarkably lower than control groups (P < 0.01). Comparing with controls and before DM treatment, sharp increases in [Ca(2+)]i concentration [(451.4 +/- 42.3), (536.9 +/- 47.5) and (870.9 +/- 100.5) nmol/L respectively] were observed when astrocytes were incubated with 1 x 10(-7), 1 x 10(-6) and 1 x 10(-5) mol/L DM for 5 minutes (P < 0.01). After astrocytes were treated with 1 x 10(-8), 1 x 10(-7), 1 x 10(-6), 1 x 10(-5) mol/L DM for 15 minutes, the [Ca(2+)]i concentrations were decreased to (124.3 +/- 6.0), (131.3 +/- 19.1), (118.9 +/- 1.4), (136.6 +/- 3.8) nmol/L respectively, which were significantly different from those of controls and before treatment. And this situation was almost keeping stable to 30 min. CONCLUSION: The cell survival rate was decreased and the [Ca(2+)]i concentration was temporarily increased when astrocytes were treated with DM. PMID- 14761487 TI - [Effects of organophosphates on cell proliferation and 45Ca uptake of neuroblastoma SH-SY5Y cells]. AB - OBJECTIVE: To explore the mechanism of cytotoxic effects of the organophosphates (OPs) with delayed neurotoxicity on human neuroblastoma cells. METHODS: The proliferation of neuroblastoma SH-SY5Y cells was determined by MTT spectrometry. (45)Ca uptake was determined by adding (45)CaCl(2) and tri-o-cresyl phosphate (TOCP) or methamidophos into the cultured medium for the SH-SY5Y cells. The cells were incubated and then lysed and finally counted in a Beckman LS 6000 liquid scintillation spectrometer. RESULTS: Methamidophos stimulated the cell proliferation of SH-SY5Y at its lower concentrations (7 x 10(-7) mol/L to 7 x 10( 6) mol/L), with an increase by 28% at 7 x 10(-7) mol/L; however, it inhibited the proliferation at higher ones (7 x 10(-4) mol/L to 7 x 10(-3) mol/L) with 62% inhibition at 7 x 10(-3) mol/L. TOCP only inhibited the cell proliferation at high concentration (with 34% inhibition at 7 x 10(-3) mol/L) and markedly inhibited calcium uptake of the cells up to 55% at higher concentrations (1 x 10( 6) mol/L to 1 x 10(-4) mol/L); while the uptake was stimulated by OPs up to 241% of increase at lower concentrations (1 x 10(-9) mol/L to 1 x 10(-7) mol/L). CONCLUSION: The interference of growth in nerve cells and disturbance of calcium homeostasis may be involved in the mechanisms of neurotoxicity of OPs. PMID- 14761486 TI - [Effects of pyrethroids on the activity of gamma-aminobutyric acid transferase in rat brain]. AB - OBJECTIVE: To study the effects of pyrethroids on the activity of gamma aminobutyric acid transferase (GABAT) in rat brain. METHOD: The coupled enzyme ultraviolet spectrophotography was applied to observe the effects of deltamethrin (DM) and permethrin (PM) on the activities of GABAT in rat cerebral cortex, hippocampus, corpus striatum and cerebellum in vitro and in vivo. RESULTS: In vitro, DM and PM had no significant effects on the activities of GABAT in rat cerebral cortex, hippocampus, corpus striatum and cerebellum at the final concentration of 10(-9) - 10(-4) mol/L. When 37.5 mg/kg DM and 600 mg/kg PM were orally administrated to the rats at one time, the activities of GABAT in rat cerebral cortex, hippocampus and cerebellum in the DM group [(2.96 +/- 0.43), (2.13 +/-0.44), (5.12 +/- 1.36) nmol x mg pro(-1) x min(-1), respectively] were lower than those in the control group [(3.43 +/- 0.41), (2.68 +/- 0.47), (6.74 +/ 1.64) nmol x mg pro(-1) x min(-1)] (P < 0.05), and the activities of GABAT in rat cerebral cortex and hippocampus in the PM group [(4.57 +/- 0.30), (4.18 +/- 0.63) nmol.mg pro(-1) x min(-1), respectively] were higher than those in the control group (P < 0.05). When 12.5 mg/kg DM and 200 mg/kg PM were orally administrated to the rats once a day for consecutive five days, the two pesticides had no significant effects on the activities of GABAT in rat cerebral cortex, hippocampus, corpus striatum and cerebellum (P > 0.05). CONCLUSIONS: In vitro, DM and PM had no significant effects on the activity of GABAT in rat brain; in vivo, DM and PM may have different effects on the activity of GABAT in rat brain, which deserve further study. PMID- 14761488 TI - [Tobacco "pneumoconiosis"]. PMID- 14761489 TI - [Effects of treatment with redox agent, blood transfusion and scopolamine on 200 patients with severe chlorphenamidine poisoning]. AB - OBJECTIVE: To verify the effects of treatment with blood transfusion and scopolamine on severe chlorphenamidine poisoning (SCP). METHODS: 400 patients with severe oral chlorphenamidine poisoning were randomly divided into two groups. 200 patients (Group I) were treated with the traditional combined therapy including gastrolavage, purgation and taking redox agent (methylene blue and vitamin C) while the other 200 patients (Group II) in addition to the above mentioned therapy, received blood transfusion and scopolamine injection. RESULTS: The cure rate of Group II was 99.5% and significantly higher than that of Group I (91.0%, P < 0.01). The average time of improving in health in Group II [(8.71 +/- 1.49) h] was obviously shorter than those in Group I [(10.65 +/- 1.72) h, P < 0.01]. Blood methemoglobin concentrations in Group II at 3, 7, 12, 24 h after admission [(43.58 +/- 2.69), (34.21 +/- 2.30), (20.60 +/- 4.03), (13.50 +/- 1.65) g/L respectively] were obviously lower than those in Group I [(54.42 +/- 12.79), (42.17 +/- 22.34), (30.66 +/- 17.67), (19.01 +/- 0.61) g/L respectively, P < 0.01]. CONCLUSION: Blood transfusion and scopolamine had distinctive therapeutic effect on SCP to makeup the deficiency of redox agent. Combination of three therapies may potentiate the detoxication for chlorphenamidine. PMID- 14761490 TI - [Application of T grain technique to the diagnosis of lung disease and analysis of its image quality]. AB - OBJECTIVE: To compare the image quality of T grain green sensitive film (TML-1) and Lanex Gd(2)O(2)S rare earth intensifying screen with that of XK-1 blue sensitive film and calcium tungstate (CaWO(4)) intensifying screen, and to study the application of T grain technic to the diagnosis of lung diseases. METHODS: 160 coal miners were randomly selected to take both TML-1 and XK-1 chest film of high kV radiographs at the same time. Silver halide granule, fluorescence of intensifying screen, radiographic parameters, the density at different points in the lung and chest radiographs were observed. RESULTS: Silver grains in TML-1 film were more homogeneous in distribution than in XK-1 film. Luminous intensity of Lanex Gd(2)O(2)S rare earth intensifying screen was brighter than CaWO(4) intensifying screen in the same exposure. The exposure doses of TML-1 film was reduced to one third of XK-1 film. The density of chest radiographs was 0.24 to 2.74 in TML-1 film, and 0.30 to 2.60 in XK-1 film. There were greater exposure latitude and more informations in TML-1 film. By apertured-disc observation, the fine structure of lung in TML-1 film was clearer than in XK-1 film, the shape was more concrete and reliable, visualizability was stronger. CONCLUSION: T grain technique may obviously improve the clearness and resolution of image, and enhance the transmission of information, as well as increase the diagnostic informations. PMID- 14761491 TI - [Effect of modified ricin on the reduction of hepatotoxicity in mice]. AB - OBJECTIVE: To observe the significance of ricin (RT) with chemical nidification to reduce the hepatotoxicity in mice and its anticancer effect. METHODS: Mice were exposed to RT and RT-PDP [ricin chemically modified by N-succinimidyl 3-(2 pyridyldithio)-propionate, (SPDP)] respectively, and their serum activity of glutathione-S-transferase (GST) and liver glutathione (GSH) content were determined. The ultramicro-structure under electron microscope was also observed. RESULTS: The GST activity increased with doses, and the increase in ricin group was higher than that in RT-PDP group; the activities of GST in RT group at 12.5, 15.0 micro g/kg [(93.65 +/- 12.30), (153.71 +/- 26.64) IU/L respectively] were higher than those in RT-PDP group [(62.97 +/- 11.22), (78.20 +/- 15.71) IU/L] (P < 0.05). The contents of GSH were decreased with doses; but the contents of GSH in RT-PDP group at 2.5, 5.0, 7.5, 10.0, 15.0 micro g/kg [(6.34 +/- 1.43), (4.14 +/- 1.82), (3.54 +/- 0.64), (2.73 +/- 1.82), (1.82 +/- 0.62) micro mol/L respectively] were still higher than those in RT group [(3.53 +/- 0.95), (2.12 +/ 0.54), (1.82 +/- 0.71), (1.52 +/- 0.34), (0.81 +/- 0.36) micro mol/L] (P < 0.01). Electron microscopic examination showed that the injury of liver cells in RT group was more severe than that in RT-PDP group. CONCLUSION: The hepatotoxicity of ricin in mice may be reduced by chemical modification. PMID- 14761492 TI - [The mechanism of JWA gene involved in oxidative stress of cells]. AB - OBJECTIVE: To study the mechanism of gene JWA involved in oxidative stress under hydrogen peroxide (H(2)O(2)) exposure. METHODS: Both MCF-7 (human breast cancer cell line) and WI-38 (human embryo lung fibroblast cell line) cells were treated with 1 mmol/L of H(2)O(2) with or without pre-incubation of taurine (tau). Malondialdehyde (MDA) and glutathione (GSH) contents in supernatant of cell culture were measured; RT-PCR and Western blotting were carried out for evaluation of the expressions of JWA mRNA and protein respectively. Heat shock proteins (HSP27, HSP70 and HSP90) were also analyzed. RESULTS: The contents of MDA before and after H(2)O(2) treatment in MCF-7 cells were (0.531 +/- 0.038), (0.674 +/- 0.410) mmol/L respectively, (P < 0.01), while those in WI-38 cells were (0.572 +/- 0.035), (0.683 +/- 0.028) mmol/L respectively, (P < 0.01). The contents of GSH before and after H(2)O(2) treatment in MCF-7 cells were (0.053 +/ 0.002), (0.044 +/- 0.002) g/L respectively, (P < 0.01), while those in WI-38 cells were (0.058 +/- 0.002), (0.050 +/- 0.002) g/L respectively, (P < 0.01). The expression of JWA mRNA was down regulated, at 6 h it decreased by 68.4%, while in WI-38 cells no obvious change found. JWA protein and HSP27 showed markedly increased after H(2)O(2) treatment in both cells but not in similar extent. CONCLUSION: Oxidative stress signal pathways of JWA gene varied between cancer and non-cancer cell lines; JWA protein may have a similar function as HSP27 and act as an important signal molecule in H(2)O(2) induced cell injury. PMID- 14761493 TI - [Recent study in relationship of cytokines to etiology of Pneumoconiosis ]. PMID- 14761494 TI - [Progresses in studies of cellular mechanism of pulmonary fibrosis]. PMID- 14761495 TI - [Mechanism and effect of taurine on pulmonary fibrosis]. PMID- 14761496 TI - [Investigation of pneumoconiosis in a factory ]. PMID- 14761497 TI - [Blood lipids and lipoproteins in acute organophosphorus insecticide poisoning patients]. PMID- 14761499 TI - [Acute poisoning in freight workers]. PMID- 14761498 TI - [Ginkgo biloba extract protection in acute paraquat poisoning of rat lung tissue ]. PMID- 14761500 TI - [Effects of organophosphate insecticide on blood nitric oxide in rabbits]. PMID- 14761501 TI - [Effect of sea water submersion on heart in rats ]. PMID- 14761502 TI - [Investigation of hearing in noise exposed workers in a factory]. PMID- 14761503 TI - [Experiment on chromosome aberration in human lymphocytes due to arsenic exposure]. PMID- 14761504 TI - [4 cases of pneumonectomy in silicosis misdiagnosed as lung cancer]. PMID- 14761505 TI - [Effect of salmeterol on function of bronchi in pneumoconiosis of coal miners]. PMID- 14761506 TI - [Intravenous immunoglobulin infusion treatment for pneumoconiosis complicated with severe asthma in coal miners ]. PMID- 14761507 TI - [11 cases of treatment of acute food poisoning ]. PMID- 14761508 TI - [Blood transfusion for the treatment of poisoning]. PMID- 14761509 TI - [46 cases of aplastic anemia caused by benzene]. PMID- 14761510 TI - [Regulation of acute occupational poisoning conditions in 27 companies ]. PMID- 14761511 TI - [Study of methods of determining amount of airborne thionyl chloride in the workplace]. PMID- 14761514 TI - [The relationship between excitatory amino acids and acute intoxicated encephalopathy induced by 1,2-dichloroethane]. AB - OBJECTIVE: To explore the relationship between excitatory amino acids (EAAs) and acute in toxicated encephalopathy induced by 1,2-dichloroethane (1,2-DCE). METHODS: SD rats were randomly divided into 7 groups: 1 control group, 3 exposure (5.0, 10.0, 20.0 g/m(3)) groups and 3 duration (2, 4, 6 h after 10.0 g/m(3) exposure) groups. Rats were exposed to 1,2-DCE for 12 h by continual static inhalation. Water content of cerebral cortex and medulla was measured by wet-dry method; The contents of glutamate (Glu), aspartate (Asp), glycine (Gly) and gamma aminobutyric acid (GABA) in brain tissues were determined by high performance liquid chromatography (HPLC). RESULTS: Water content of cerebral cortex in 3 exposure groups (76.10% +/- 1.41%, 76.45% +/- 0.75%, 79.95% +/- 1.45% respectively) were higher than that of control group (74.22% +/- 1.77%, P < 0.05, P < 0.01). That of medulla was increased significantly merely at 20.0 g/m(3) (71.77% +/- 3.07%, P < 0.05). Water content of cortex in 3 duration groups (79.36% +/- 2.10%, 79.48% +/- 1.21%, 80.64% +/- 1.93% respectively) were higher than that of 10.0 g/m(3) instant exposure group (P < 0.05). The content of Asp [(4.83 +/- 0.35) micro mol/g, (7.17 +/- 0.40) micro mol/g, (10.52 +/- 0.39) micro mol/g], Glu [(23.86 +/- 0.62) micro mol/g, (31.21 +/- 2.50) micro mol/g, (28.23 +/- 1.58) micro mol/g] and Gly [(5.59 +/- 1.01) micro mol/g, (6.06 +/- 0.83) micro mol/g, (7.26 +/- 1.34) micro mol/g] in exposure groups were higher than those of corresponding control groups [(3.72 +/- 0.48) micro mol/g, (21.09 +/- 1.20) micro mol/g, (3.83 +/- 0.44) micro mol/g, P < 0.05, P < 0.01]. Compared to 10.0 g/m(3) instant group, Asp content was increased at 2, 4, 6 h (P < 0.01), Glu content at 2, 4 h (P < 0.05), and peak value of Glu appeared at 4 h [(35.40 +/- 2.40) micro mol/g] while Gly content was significantly decreased (P < 0.01). GABA did not show evident changes in both exposure or duration groups (P > 0.05). CONCLUSION: EAAs appears to be related with the development of acute intoxicated encephalopathy induced by 1,2-DCE, and its damage to neuron might be one of the mechanisms of brain edema. PMID- 14761515 TI - [Effect of the polymorphism of myeloperoxidase gene on the risk of benzene poisoning]. AB - OBJECTIVE: To study the relationship between the polymorphism myeloperoxidase (MPO) gene and the genetic susceptibility to benzene toxicity in workers exposed to benzene and in patients with benzene poisoning. METHODS: Using polymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLP) techniques, the genotypes' polymorphism of MPO gene in 35 patients with chronic benzene poisoning, 46 workers exposed to benzene from the same workplace (as exposed control) and 26 controls were analyzed. RESULT: There were three (G/G, G/A and A/A) genotypes in the region of 463 bp upstream of MPO gene. The distribution frequency in G/G wild-type genotype in patients was 27.4% more than that in the exposed workers. The risk of benzene-hematotoxicity in those with G/G genotype was 2.8-fold higher than G/A + A/A genotype (OR = 2.835, 95% CI: 1.065 - 7.549, P < 0.05). The polymorphism of myeloperoxidase was not associated with gender specific. CONCLUSION: In the same benzene-exposed environment, the subjects with MPO-463 G/G genotype may be more susceptible to benzene toxicity. PMID- 14761516 TI - [Study of T lymphocyte rDNA transcription activity in chronic benzene poisoning patients]. AB - OBJECTIVE: To observe the changes in peripheral blood T lymphocyte rDNA transcription activity and to study the significance of immune monitoring for patients with chronic benzene poisoning. METHODS: Venous blood samples were withdrawn from 39 patients with chronic benzene poisoning, 20 patients with malignant disease and 22 healthy controls. Cell culture, argyrophil staining method, I-CLQ cell image analysis system were used in this study. rDNA transcription activity which was expressed by the ratio of integrated area (IA) of nucleolus to that of nucleus, and the ratio of integrated optical density (IOD) of argyrophilic nucleolus to that of argyrophilic nucleus. RESULTS: (1) The value of IA and IOD in chronic benzene poisoning patients (7.95% +/- 1.13% and 7.15% +/- 1.15% respectively) were lower than those in controls (9.59% +/- 1.26% and 8.92% +/- 1.18% respectively), P < 0.01. The value of IA and IOD in chronic moderate benzene poisoning patients (6.54% +/- 0.88%) and (5.47% +/- 0.80%) were lower than those to be observed (7.98% +/- 1.06% and 7.13% +/- 0.96% respectively) as well as in mild poisoning patients (8.19% +/- 1.06% and 7.44% +/ 1.06% respectively), P < 0.05. (2) The value of IA and IOD in malignant group (4.10% +/- 1.50%, 3.67% +/- 1.42%) were much more lower than those in controls and in chronic benzene poisoning patients (P < 0.01). CONCLUSION: rDNA transcription activity may be an index to monitor the cellular immune function of chronic benzene poisoning patients. PMID- 14761518 TI - [A study on biomarkers of styrene]. AB - OBJECTIVE: To study the biomarkers of styrene and to provide theoretical basis for bio-monitoring of styrene. METHODS: Urinary mandalic acid (MA), phenylglyoxalic acid (PGA) and mercapturic acid (MUA) of styrene were examined by high performance liquid chromatography (HPLC). RESULTS: The correlation regression equations between exposure dose and MA, PGA and MUA level in morning urinary samples were: y = 2.58x + 70.82; y = 1.66x + 37.42; y = 0.05x + 0.55 respectively. The correlation regression equations between exposure dose and MA, PGA and MUA level in post-shift urinary samples were: y = 1.85x + 89.02; y = 1.33x + 4.32; y = 0.04x + 0.68 respectively. All showed close dose-response relationship. CONCLUSIONS: The level of MA, PGA and MUA in morning or post-shift urinary samples may be used as bio-monitoring indexes of styrene. PMID- 14761520 TI - [Effects of acrylamide on DNA damage in human keratinocytes]. AB - OBJECTIVE: To investigate the toxic and DNA damaging effect of acrylamide (AA) on human keratinocytes and its mechanism. METHODS: (1) After the keratinocyte cell line HaCaT cells were exposed to AA with different concentrations for 44 hours, cell survival rate was detected by MTT method. (2) The effects of DNA damage of exposed cells were detected by comet assay. (3) After treating the cells with 2.00 mmol/L of AA plus 0.50 mmol/L of 1-aminobenzotriazole (1-ABT), an inhibitor of cytochrome P-450 enzymes (CYP-450), for 4 hours, the relationship between DNA damage and CYP-450 was studied. RESULTS: (1) Cytotoxicity measurement of AA showed that cell survival rate decreased significantly after 44-hour treatment. (2) Cytotoxicity was not detected after 4-hour AA treatment, but significant DNA damage was observed in all treatment groups, and the degree of damage increased with the concentration of AA. Moreover, the tail lengths of comet cells were in dose-effect relationship. As for cells treated by 1-ABT with 2 mmol/L AA, comet rate and tail length were 15.4% and (8.2 +/- 2.0) micro m respectively, which were decreased significantly (P < 0.01) when compared with 2 mmol/L AA treatment group [80.6% and (44.3 +/- 4.0) micro m]. CONCLUSIONS: Acrylamide has significant cytotoxicity and genotoxicity on HaCaT cells. AA-induced DNA damage may be related to the oxidative metabolite(s) of AA through CYP-450. PMID- 14761521 TI - [Study on mitochondrial DNA damage in peripheral blood nucleate cells of the workers exposed to acrylonitrile]. AB - OBJECTIVE: To study the potential aging effect on workers exposed to acrylonitrile (ACN). METHODS: The deletion rates of mitochondrial DNA (mtDNA) in peripheral blood nucleate cells of 47 exposed workers and 47 non-exposed workers (as control), as well as 12 old people and 12 young people were measured with polymerase chain reaction (PCR). RESULTS: The positive rates of mtDNA deletion in peripheral blood nucleate cells were 17.02% in the workers exposed to ACN and 25.00% in group of old people. However, the mtDNA deletion was not detected in the control group and young people. CONCLUSIONS: ACN could induce mtDNA deletion in peripheral blood nucleate cells of the exposed workers. There may be a potential molecular effect of occupational ACN exposure on workers' aging. PMID- 14761523 TI - [Studies on fluoride concentration and cholinesterase activity in rat hippocampus]. AB - OBJECTIVE: To study the accumulation of fluoride in rat hippocampus and its effect on cholinesterase activity. METHODS: Rats were subchronically exposed to NaF, and fluoride concentration and cholinesterase activity in rat hippocampus were determined. RESULTS: Fluoride concentration in rat hippocampus was significantly correlated with the dosage of fluoride, and there were significant differences among high dosage group [(13.03 +/- 1.79) micro g/g], low dosage group [(9.83 +/- 0.92) micro g/g] and control [(8.27 +/- 1.11) micro g/g], P < 0.01. Acetylcholinesterase activities among three groups [(0.111 +/- 0.031) micro mol/mg, (0.143 +/- 0.025) micro mol/mg, (0.183 +/- 0.027) micro mol/mg] were also significantly different (P < 0.01), which was negatively correlated with fluoride concentration in rat hippocampus (r = -0.700, P < 0.01). The activity of butylcholinesterase in high dosage group [(0.041 +/- 0.010) micro mol/mg] was different from that of control [(0.067 +/- 0.025) micro mol/mg, P < 0.05], but the activity was not significantly related with fluoride concentration in rat hippocampus (r = -0.317, P = 0.094). CONCLUSION: Fluoride may go through the blood-brain barrier and accumulate in rat hippocampus, and inhibit the activity of cholinesterase. PMID- 14761525 TI - [Construction of double-strand break repair protein hKu70 deficient cell strain and its biologic characters]. AB - OBJECTIVE: To construct DNA double-strand break (DSB) repair protein hKu70 deficient cell strain and to observe its biological characters for studying the functions of hKu70 gene and the effects of occupational harmfulness factors on DSB repair. METHODS: Human lung fibroblasts (HLF) were transfected with the eukaryotic expression plasmids of hKu70 gene antisense RNA (pEGFP-C1-K) to construct hKu70 protein deficient cells (named as "HLFK"). The protein expression levels of hKu70 gene in HLFC and HLFK were detected by the Western blotting to estimate the effects of antisense inhibition. Morphology, growth character and growth status in soft agar of transfected HLFK were observed. RESULTS: pEGFP-C1-K vector was successfully expressed in HLF. The protein expression level of hKu70 gene in HLFK was decreased by 42% as compared with that in HLFC. No obvious changes of the biologic characters were observed in HLFK. CONCLUSION: The hKu70 protein deficient cell strain was successfully constructed. The hKu70 protein deficiency alone didn't induce obvious changes of the biological characters in HLFK. PMID- 14761526 TI - [Effect of L-methionine on trace elements in lead-intoxicated mice]. AB - OBJECTIVE: To investigate the effect of L-methionine (L-Met) on the content of Zn, Cu, Mn, Fe in liver, brain, spleen and kidney of lead intoxicated mice. METHODS: Distilled water was given to 10 mice (normal control group) and lead acetate solution of 400 micro g/ml Pb(2+) to 20 mice to serve as drinking water for 10 days. The lead administration was then withdrawn and lead exposed mice were randomly divided into two groups: the lead control group took distilled water as drinking water for 4 weeks to serve as positive control, the other one took L-Met solution (0.5 mg/ml) as drinking water for 4 weeks (Pb + L-Met group) to serve as the treatment group. All the animals were sacrificed on the 1st day after 4 weeks, and the contents of Zn, Cu, Fe, Mn, Pb in liver, brain, spleen and kidney were measured by Inductively Coupled Plasma (ICP) Emission Spectrometry. RESULTS: Lead contents in liver, brain, spleen and kidney of Pb control group [(1.490 +/- 1.654) micro g/g, (3.470 +/- 2.757) micro g/g, (4.975 +/- 2.993) micro g/g, (0.066 +/- 0.001) micro g/g respectively], were higher than those in normal control group [(0.015 +/- 0.001) micro g/g, (0.009 +/- 0.007) micro g/g, (0.027 +/- 0.002) micro g/g, (0.006 +/- 0.015) micro g/g, P < 0.05] while Zn contents in liver, brain, spleen and Fe and Mn content in liver, brain, spleen and kidney in Pb control group were lower than those in normal control group (P < 0.05). Pb contents of brain, spleen and Cu content of kidney in Pb + L-Met group were higher than those in normal control group (P < 0.05). Zn contents of liver, brain, spleen, Fe contents of liver, brain, spleen, kidney, and Mn contents of brain, spleen in Pb + L-Met group were lower than those in normal control group (P < 0.05). Fe contents of liver, brain, Zn content of spleen, Cu content of kidney and Mn contents of liver, brain, spleen in the Pb + L-Met group were higher than those in the Pb control group (P < 0.05). The lead levels of four organs in the Pb + L-Met group were lower than those in the Pb control group (P < 0.05). CONCLUSION: Lead could be eliminated by L-Met, which may affect the distribution and metabolism of trace elements in mice. PMID- 14761528 TI - [Effects of lead on thyroid function of occupationally exposed workers]. AB - OBJECTIVE: To explore the effects of lead on the thyroid function of occupationally exposed workers. METHOD: 157 workers occupationally exposed to lead in a smelting factory were investigated. The concentration of lead in air at workshop was measured by flame atomic absorption spectrophotometry (FAAS) and the levels of blood lead (PbB) by atomic absorption spectrophotometry (AAS), zinc protoporphyrin (ZPP) by ZnPP meter, as well as the indexes of thyroid function, thyroid-stimulating hormone (TSH), triiodothyronine (T(3)), thyroxin (T(4)), free T(3) (FT(3)), and free T(4) (FT(4)) in serum by radioimmunoassay. RESULTS: The workers with higher level of blood lead (> 2.88 micro mol/L) showed lower levels of T(3) [(1.54 +/- 0.39) nmol/L] and FT(3) [(5.50 +/- 1.26) pmol/L] than those with lower blood lead level [PbB: (1.92 approximately 2.88) micro mol/L group, T(3): (1.71 +/- 0.45) nmol/L, FT(3): (6.12 +/- 1.64) pmol/L, P < 0.05]. There was no obvious effect of length of service on thyroid hormone of exposed workers. CONCLUSION: Higher level of blood lead may cause certain damage to thyroid function by inhibiting deiodination of T(4). No obvious relation between length of service and thyroid function was found. PMID- 14761530 TI - [Effect of aberrant DNA methylation on the expression of cancer-related genes in Cd-transformed cells]. AB - OBJECTIVE: To study aberrant DNA methylation potentially resulting in changes in the expression of cancer-related genes as a possible epigenetic mechanism for cadmium carcinogenesis. METHODS: Genomic DNA isolated from CdCl(2)-transformed BALB/c-3T3 cells was digested with Mse1 (methylation non-sensitive) alone or with Mse1 and BstU1 (methylation sensitive). The resulting DNA was analyzed for aberrant methylation using PCR-based technique-Methylation-Sensitive Restriction Fingerprinting (MSRF). Several DNA fragments differentially methylated in the transformed cells identified by MSRF were confirmed by Southern hybridization analysis using the aberrantly methylated DNA fragments as the probes. RESULTS: Aberrant DNA methylation was identified in the transformed cells. DNA sequencing and sequence similarity analysis identified one of the aberrantly methylated DNA fragments as the p16 tumor suppressor gene. CONCLUSION: DNA hypermethylation is known to result in gene silencing, it appears that hypermethylation of p16 gene may represent a possible epigenetic mechanism for Cd-induced cell transformation and carcinogenesis. PMID- 14761531 TI - [Preliminary study on structure and chemical characteristics of deoxyguanoside benzoquinone adducts]. AB - OBJECTIVE: To detect the structure and chemical characteristics of the adduct from the reaction of p-benzoquinone (BQ) with deoxyguanoside (dGMP). METHODS: dGMP and calf thymus DNA were reacted with BQ in buffered solutions with neutral pH, the reaction products were separated and purified by high performance liquid chromatograph (HPLC), and then characterized by UV spectroscopy and mass spectrometry. RESULTS: The reaction of BQ with dGMP yielded two adduct products (Ad(1) and Ad(2) respectively). The characterized results of Ad(1) suggested that BQ reacted at the N-1 and N(2) position of dGMP by losing one H(2)O molecule, the molecular weight of Ad(1) was 437, and the molecular formula was C(16)H(16)O(8)N(5)P. Ad(1) could also be detected from calf thymus DNA reacted with BQ in vitro, which possessed the same elution profile by HPLC analysis. Meanwhile, Ad(2) was detected in the experimental condition. It was proposed that Ad(2) was formed by BQ reacted at the N-9 position of dGMP by losing one molecule of deoxyribose, the molecular weight was 241, and the molecular formula was C(11)H(7)O(2)N(5). CONCLUSION: The structure of one major adduct from reaction of BQ with DNA is (3'-OH)-1, N(2)-C(6)H(5)CH-2'-deoxyguanosine-5'-monophosphate. PMID- 14761533 TI - [Effects of noise on antioxidant enzymes of cochlea in guinea pigs]. AB - OBJECTIVE: To investigate the effect of noise on the antioxidant enzymes of cochleae. METHODS: 16 male pigmented guinea pigs (250 - 300 g) were randomly divided into 2 groups, control group and noise group. Each group had 8 animals. The animals in noise group were performed auditory evoked brainstem responses (ABR) recording before and after exposure to a continuous noise (4 kHz, octave band, 100 dB, SPL) 8 h/d for 3 consecutive days. Immediately at the end of the third day's noise exposure after ABR recording, guinea pigs were decapitated. Both the right and the left cochlea with the bony capsule removed were homogenized, and the supernatants were prepared for assays. Reactive oxygen species (ROS), superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) were measured. RESULTS: ROS level of the noise group [(281.2 +/- 3.5) U/mg pro] was significantly higher than that of the control group [(273.0 +/- 3.2) U/mg pro, P < 0.05] and SOD, CAT and GSH-Px activities of the noise group [(206.5 +/- 5.1) NU/mg pro, (47.0 +/- 9.0) U/g pro, (14.1 +/- 2.5) U/mg pro respectively] were significantly lower than that of the control group [(221.8 +/- 4.8) NU/mg pro, (60.8 +/- 9.9) U/g pro, (21.1 +/- 3.1) U/mg pro respectively, P < 0.05]. CONCLUSION: Noise may damage the defensive system of antioxidant enzymes in cochlea. PMID- 14761534 TI - [Protective effect of Gastrodia elata and E-gelatin on lead-induced damage to the structure and function of rat hippocampus]. AB - OBJECTIVE: To observe the counteraction of Gastrodia elata and E-gelatin on the effect of subchronic lead poisoning on the ability of learning and memory and the ultrastructure in hippocampus. METHODS: Subchronic lead acetate exposure was given to rats (0.2 g x kg(-1) x d(-1)). Single and combined administration of Gastrodia elata (4 g x kg(-1) x d(-1)) and E-gelatin (1 g x kg(-1) x d(-1)) were conducted at the same time. Pb concentration in blood, and the ability of learning and memory (Y-maze test) of rats were measured. Ultrastructure of CA3 pyramidal cells in hippocampus under transmission electron microscope was observed. RESULTS: Blood Pb concentrations in each group (Pb group: 690.6 micro g/L, Pb + Gastodiae eleta group: 688.8 micro g/L, Pb + E-gelatin group: 663.8 micro g/L, Pb + combined group: 667.2 micro g/L) were higher than that in the control (28.24 micro g/L, P < 0.01). But there was no significant difference (P > 0.05) among these groups. In Y-maze test, the number of electric stroke in Pb group is higher than that in control (P < 0.01). Gastrodia elata or E-gelatin used singly could significantly reduce the number of electric stroke of lead exposed groups (P < 0.05 in the first month and P < 0.01 in the second and third month). And the effect of combined use of them was more efficient than single use (P < 0.01). Under electron microscope, no anomaly was seen in the pyramidal cells of CA3 area in hippocampus of control group. But there was significant anomaly such as neucleus separation in the cells of Pb exposed group. In Pb-Gastrodia elata group, there were some stress response phenomena such as the occurrence of huge mitochondria while in Pb-E-gelatin group, the anomaly was mild. The cells in the CA3 area in hippocampus of the pb-Gastrodia-E-gelatin group were almost normal, and also showed some stress response phenomena. CONCLUSION: Gastrodia elata and E-gelatin may protect the neurons in CA3 area of hippocampus against damage induced by lead to improve the ability of learning and memory, and they have synergistic effect. PMID- 14761536 TI - [The antagonistic action of Gastrodia elata combined with E-gelatin on lead induced down regulation of c-fos expression in rat brain]. AB - OBJECTIVE: To observe the effect of lead on the expression of c-fos mRNA in hippocampus and cerebellum and the ability of learning and memory in rat, and to observe the antagonistic action of Gastrodia elata and E-gelatin on the effect mentioned above. METHODS: Rats were exposed to lead acetate (0.2 g x kg(-1) x d( 1) and 0.1 g x kg(-1) x d(-1)). The single and combined administration of Gastrodia elata (4 g x kg(-1) x d(-1)) and E-gelatin (1 g x kg(-1) x d(-1)) were conducted at the same time. Blood lead concentration in rats were measured. The ability of learning and memory by Y-maze test were examined. The expression of c fos mRNA in hippocampus and cerebellum during Y-maze test were observed by in situ hybridization (ISH). RESULTS: (1) Blood lead concentrations were significantly increased in both high and low doses of lead-exposed rats (P < 0.01). But there were no differences among high dose groups or low dose groups (P > 0.05). (2) In Y-maze test, the number of electric stroke during learning in lead-exposed group was significantly increased (P < 0.01) while that in Gastrodia elata and E-gelatin groups was significantly decreased (P < 0.05), and that in combined use group was more significantly decreased (P < 0.01). (3) There were much more deep-colored c-fos positive cells in CA3 area of hippocampus in low dose of Pb-exposed with Gastrodia elata + E-gelatin group. These cells were also found in high Pb with combined use or single use group, but which were not so densely distributed as the former. And they were hardly found in high and low doses of Pb alone groups. The changes of expression of c-fos cells in cerebellam were similar to those in hippocampus. CONCLUSION: The down regulation of c-fos expression may be one of the molecular mechanism of lead-induced impairment of learning and memory. Gastrodia elata and E-gelatin may antagonize the effect of lead on c-fos expression, and combined use of both drugs may potentiate the antagonism. PMID- 14761537 TI - [Effects of power frequency magnetic field on gap junction intercellular communication of astrocytes]. AB - OBJECTIVE: In order to explore if power frequency magnetic field (PFMF) can act as cancer promoter or be synergistic with phorbol 12-myristate 13-acetate (TPA) in cancer promotion, the effects of 50 Hz MF on gap junction intercellular communication (GJIC) of astrocytes were observed. METHODS: Fluorescence redistribution after photobleaching (FRAP) was adopted to observe the recovery of fluorescence intensity in the bleached cells thus to estimate intercellular communication by gap junction. Comparative fluorescence intensity recovery rate (CFIRR) was as evaluation index. The effects of 50 Hz MF alone or with TPA on GJIC of astrocytes were studied. RESULTS: After 3 ng/ml TPA treatment for 1 hour, M(d) of CFIRR was 4.53%/min, whereas that in the control group was 9.74%/min (H = 12.084, P < 0.005). After exposure to 0.8 and 1.6 mT magnetic field for 24 hours respectively, M(d) of CFIRR was 8.25%/min and 6.68%/min respectively, no significant difference from that of control (H = 32.617, P > 0.05). After exposure to 0.8 and 1.6 mT magnetic field for 23 hours then combined with 3 ng/ml TPA treatment for 1 hour, M(d) of CFIRR was 3.32%/min and 2.85%/min respectively, also no significant difference from that in the group treated with 3 ng/ml TPA alone (H = 2.589, P > 0.05). CONCLUSION: 50 Hz MF (within 0 - 1.6 mT) alone could not inhibit GJIC of astrocytes; with TPA, could not enhance the inhibition of TPA on GJIC of astrocytes. But with MF intensity increasing, the inhibition of MF on GJIC showed elevated tendency. PMID- 14761538 TI - [Study of platelet membrane glycoprotein expression in mice with decompression sickness]. AB - OBJECTIVE: To investigate the role of expression of platelet membrane glycoprotein CD31, CD61 and CD62p in the pathogenesis of decompression sickness. METHODS: Mice were randomly divided into decompression sickness group and normal control group. The animals in decompression sickness group were exposed to 600 kPa compressed air for 60 minute, then they were rapidly decompressed to normal pressure in one minute. At 60th minute after reducing to normal pressure, the expression of CD31, CD61 and CD62p on platelet membrane in mice was measured by flow cytometry. RESULTS: The mean fluorescence intensity of CD31, CD61 and positive percentage of CD62p on platelet membrane [(18.64 +/- 1.01), (271.06 +/- 24.25), (4.48% +/- 0.43%) respectively] in decompression sickness group were significantly increased compared with normal control group [(16.89 +/- 1.69), (234.09 +/- 15.96), (3.00% +/- 0.66%) respectively] (P < 0.05, P < 0.01). CONCLUSION: Inadequately rapid decompression may induce up regulation of platelet membrane glycoprotein CD31, CD61 and CD62p expression in mice, which may lead to thrombosis. PMID- 14761557 TI - [Reliability and validity of the Occupational Stress Scale for Chinese offshore oil platform workers]. AB - OBJECTIVE: To evaluate the validity and reliability of the Occupational Stress Scale (OSS) for Chinese offshore oil platform workers. METHODS: A 51-item self administered questionnaire developed in the light of Cooper's questionnaire and company's special situation was used to investigate 561 subjects. RESULTS: 51 occupational stress items relating to offshore oil production were subjected to factor analysis, and nine latent factors were identified, which explained 62.5% of the total variance. According to the contents described by the items included in each factor, they were respectively defined as: "the interface between job and family/social life (factor 1)", "career and achievement (factor 2)", "safety (factor 3)", "management problem and relationship with others at work (factor 4)", "physical factors of workplace (factor 5)", "platform living environment (factor 6)", "role in management (factor 7)", "ergonomics (factor 8)" and "organization structure (factor 9)". Significant difference in the score of five factors was observed among 12 different job categories by analysis of variance. After adjusting for potential confounding factors (age, educational level), hierarchical multiple regression analysis indicated that the score of the OSS was significantly and positively correlated with the poor mental health of the workers (P < 0.01). The consistent test between OSS and each factor showed that Cronbach's alpha were 0.72 - 0.91. CONCLUSION: The OSS is a valid and reliable tool for measuring occupational stress, and can be used to explore occupational stress and its influence on health and safety problems in offshore oil workers. PMID- 14761559 TI - [A study of the effect of occupational stress on glucocorticoid receptor and immune function in dispatchers]. AB - OBJECTIVE: To study the effect of chronic occupational stress on the glucocorticoid receptor (GR) and immune function. METHODS: A cross-sectional study was conducted in 112 railway station dispatchers. Perceived job stress was assessed by means of the Chinese version of the Job Content Questionnaire. The subjects were divided into high, medium and low strain groups according to the job strain score of the questionnaire. The number of GR, percentage of T lymphocyte subpopulations, concentrations of cortisol and interleukin 2 (IL-2) in blood were measured. RESULTS: The concentrations of serum cortisol in high and medium strain groups [(295.43 +/- 79.06) and (274.34 +/- 70.08) ng/ml respectively] were higher than that in low strain group [(181.01 +/- 53.41) ng/ml, P < 0.05]. GR binding capacity in both groups (4,330.0 +/- 1,001.0, 3,971.6 +/- 966.8 specific binding/cell respectively) were smaller than that in low strain group (5,141.3 +/- 1,068.5 specific binding/cell, P < 0.05). The percentage of CD(3) T lymphocyte in high strain group was higher than that in low strain group [(50.21 +/- 10.30)% vs (56.87 +/- 15.36)%, P < 0.05], and CD(4) T lymphocyte in high and medium strain groups were significantly smaller than that in low strain group [(23.27 +/- 10.01)%, (27.06 +/- 7.47)% vs (33.31 +/- 7.77)%, P < 0.05]. In contrast, the percentage of CD(8) T lymphocytes in high and medium strain groups were significantly higher than that in low strain group [(28.16 +/- 6.47)%, (25.54 +/- 6.70)% vs (21.91 +/- 5.93)%, P < 0.05]. The levels of serum IL 2 in high and medium groups were smaller than that in low strain group [(0.77 +/- 0.05), (0.80 +/- 0.07) vs (1.05 +/- 0.12) ng/ml, P < 0.05]. Correlation analysis showed that serum cortisol level was negatively correlated with CD(8) percentage (r = -0.612, P < 0.01). CONCLUSION: Chronic occupational stress may induce rise of glucocorticoid, down-regulation of GR and inhibition on immune function. PMID- 14761560 TI - [The relationship between occupational stress and cardiovascular disease risk factor]. AB - OBJECTIVE: To explore the relationship between the occupational stress and cardiovascular disease risk factors. METHODS: The subject was 93 male freight train dispatchers, the occupational stressors, personalities, and occupational strain response were measured by using questionnaires. Blood pressure, heart rate, the concentrations of the cholesterol, triglyceride, and glycosylated hemoglobin in blood serum were also measured. RESULTS: Social support score were significantly positively related to systolic pressure (r = 0.22) and diastolic pressure (r = 0.30) (P < 0.05), while job satisfaction negatively related to them and concentration of triglyceride (r = -0.37, -0.47 and -0.28 respectively, P < 0.05, P < 0.01), and self-esteem negatively to systolic pressure (r = -0.21, P < 0.05). The differences in diastolic pressure [(78.5 +/- 13.1), (83.6 +/- 8.2), (88.1 +/- 12.3), (85.8 +/- 9.8) mm Hg, P < 0.05] among groups of social support score, body mass index (BMI) (P < 0.01) among groups of job difficulty the differences in systolic pressure [(124.9 +/- 14.4), (129.7 +/- 13.1), (118.4 +/- 16.4), (133.5 +/- 23.1) mm Hg] and diastolic pressure [(85.5 +/- 11.3), (87.0 +/- 9.8), (80.1 +/- 10.1), (88.9 +/- 12.0) mm Hg] and cholesterol level in serum [(4.96 +/- 1.18), (5.39 +/- 0.85), (5.00 +/- 1.15), (4.34 +/- 0.91) mmol/L] among groups of vulnerability to stress (P < 0.05), as well as the difference in systolic pressure and glycosylated hemoglobin level in serum among groups of competition score (P < 0.05) were all statistically significant. Stepwise regression analysis showed that job time demands and negative coping affected the change of cholesterol (R(2) > 0.05); the job relation decision latitude, social support, job difficulty, personality (self-esteem and anxiety trait) and negative coping were the predictors of smoking (R(2) > 0.05). Heart rate was related to home income and competition factor of Type A Behavior (R(2) = 0.06). CONCLUSION: The psychosocial stress aspects of work may be related to some cardiovascular risk factors. PMID- 14761561 TI - [Effect of occupational stress on mental health]. AB - OBJECTIVE: To study the effect of job psychological demands and job control on mental health and their interaction. METHODS: 93 male freight train dispatchers were evaluated by using revised Job Demand-Control Scale and 7 strain scales. Stepwise regression analysis, Univariate ANOVA, Kruskal-Wallis H and Modian methods were used in statistic analysis. RESULTS: Kruskal-Wallis H and Modian methods analysis revealed the difference in mental health scores among groups of decision latitude (mean rank 55.57, 47.95, 48.42, 33.50, P < 0.05), the differences in scores of mental health (37.45, 40.01, 58.35), job satisfaction (53.18, 46.91, 32.43), daily life strains (33.00, 44.96, 56.12) and depression (36.45, 42.25, 53.61) among groups of job time demands (P < 0.05) were all statistically significant. ANOVA showed that job time demands and decision latitude had interaction effects on physical complains (R(2) = 0.24), state anxiety (R(2) = 0.26), and daytime fatigue (R(2) = 0.28) (P < 0.05). Regression analysis revealed a significant job time demands and job decision latitude interaction effect as well as significant main effects of the some independent variables on different job strains (R(2) > 0.05). CONCLUSION: Job time demands and job decision latitude have direct and interactive effects on psychosomatic health, the more time demands, the more psychological strains, the effect of job time demands is greater than that of job decision latitude. PMID- 14761563 TI - [Effect of occupational stress on cardiovascular function of different vocational population]. AB - OBJECTIVE: To study the effect of occupational stress on cardiovascular function of different vocational population. METHODS: The occupational stressors, risk factors of cardiovascular diseases were investigated by questionnaire in 839 people with 4 kinds of jobs. Blood pressure, sugar, and lipid were detected at the same time. RESULTS: Blood pressure were higher in the groups of old age, long standing and teachers, and the abnormal rate of blood pressure was 21.69%. There was no difference in abnormal ECG among ages, standing and occupation, and the abnormal rate of ECG was 19.07%. Job control, job demands, job responsibility, role in a job and shift work were the main stress factors affecting systolic and diastolic blood pressure. More conflict in job, less chance of participation, severe job loads were the risk factors of primary hypertension. Accident due to job responsibility, job responsibility, role in a job were the main risk factors of abnormal electrocardiograph. Self-respect and activity beyond work were the good modifiers of heart function. CONCLUSION: Occupational stress has certain effect on cardiovascular function. PMID- 14761564 TI - [The effects of three kinds of biochemical indices on evaluation of occupational stress]. AB - OBJECTIVE: To study the feasibility of monoamine neurotransmitters, serum glucose, serum glycerinate and cholesterol as objective indices for evaluating occupational stress. METHODS: Job stressors, modifiers, job strains in 844 people with four kinds of occupation were investigated, and the concentration of monoamine neurotransmitters, glucose, glycerinate and cholesterol in blood were detected at the same time. The methods of multiple stepwise regression and covariance analysis were used to analyze the data. RESULTS: There was close relationship between monoamine neurotransmitters and job stressors, the forecast of the equation of 5-hydroxytryptamine (5-HT), dopamine (DA) and norepinephrine (NE) was 0.7238, 0.5703, 0.4438 respectively, the critical values of them were 804.00, 226.00 and 275.00 ng/ml respectively. There was a little contribution of job stressors to the equation of glucose, glycerinate and cholesterol, the critical values were 6.40, 2.51 and 5.92 mmol/L respectively. CONCLUSION: Monoamine neurotransmitters may be a direct objective evaluating indices. Sugar, glycerinate and cholesterol may be an indirect objective indices. PMID- 14761566 TI - [The psychological effect of minesweeping infrasonic field]. AB - OBJECTIVE: To investigate the effect of infrasonic field of minesweeper on psychology of minesweeper crews. METHODS: An experimental ship was selected to conduct infrasonic minesweeping, a control ship of the same type was selected to operate likewise with the infrasound generator turned off, and another group of coast-servicemen was chosen as blank control. Attention-span test, digit memory test, two-digital numbers list and profile of mood states (POMS) were used to test the ship crews and coast-servicemen. RESULTS: There were no significant differences in psychological parameters between the two ship crews before the experiment. But the scores of two-digit figures list in ship crews were significant lower than that in coast-servicemen, (31.2 +/- 11.8, 36.4 +/- 14.5 respectively vs 45.8 +/- 13.9, P < 0.05). POMS showed that the scores of anger hostility, fatigue-inertia, and confusion-bewilderment in both ship crew groups [(15.5 +/- 6.4, 18.3 +/- 6.8), (12.1 +/- 5.0,12.3 +/- 4.9), (11.6 +/- 4.4, 12.5 +/- 4.8), respectively] were higher than those in coast-servicemen (13.9 +/- 7.0, 7.6 +/- 4.1, 8.2 +/- 4.3, respectively, P < 0.05 or 0.01), while vigor-activity in experimental group (15.0 +/- 5.9) was lower than that in coast-servicemen (19.7 +/- 4.7). After the experiment, the scores of experimental crews in digit memory test (14.5 +/- 5.0), vigor-activity (12.2 +/- 5.8), fatigue-inertia (15.8 +/- 6.2) were significantly different from those of control crews (17.3 +/- 4.8, 16.5 +/- 4.6, 11.3 +/- 5.1). CONCLUSIONS: The infrasonic field from minesweeper may injure the crew's psychological health and some neurobehavioral function. PMID- 14761568 TI - [Effect of hyperthermia combined with trauma on serum nitric oxide and mean arterial pressure in rabbits]. AB - OBJECTIVE: To study the early change of serum nitric oxide (NO) after acute heat exposure with trauma and the effect of NO on mean arterial pressure (MAP), thus to provide theoretical basis for studying the mechanism of NO effect in acute stress. METHODS: The rabbit model of acute heat exposure combined with trauma was established. The animals were divided into four groups, including control, trauma, hyperthermia and hyperthermia combined with trauma. The levels of NO were measured at different time points: 0 h, 1 h, 2 h and MAP was monitored throughout the whole experiment. RESULTS: The concentration of NO declined at first and then increased at 1 h or so after acute heat exposure and trauma. The levels of NO in hyperthermia with trauma group at 1 h, 2 h were (42.75 +/- 8.24), (59.54 +/- 9.05) micro mol/L respectively (P < 0.05), while those in control group were (56.63 +/- 3.79) and (55.22 +/- 7.15) micro mol/L, the difference at 1h between two groups was significant (P < 0.05). Under the circumstance of hyperthermia and trauma, the level of MAP declined to the lowest point at 60 - 70 min and then showed a transient rise, after that, the level declined rapidly. CONCLUSIONS: At the early stage of acute heat exposure and trauma, the concentration of serum NO declined at first and then increased, and had certain relationship with the change of MAP. PMID- 14761569 TI - [A cohort study on the relationship between blood pressure levels and the mortality of cerebro-cardiovascular diseases in Guangzhou workers]. AB - OBJECTIVE: To study the relative risk (RR) of mortalities of cerebro cardiovascular diseases (CVD) in Guangzhou workers with hypertension. METHOD: Prospective cohort study was conducted in 78,379 workers, aged >or= 35, from 399 factories. Cox regression model were mainly used for data analysis. RESULTS: (1) There were 48,705 male and 29,674 female workers in the cohort. All workers were followed-up for 8 years. 363 CVD deaths (male 305, female 58) mainly died of stroke, coronary heart disease and hypertension. The crude mortalities were 78.58/100,000 person years in male and 24.55/100,000 person years in female. (2) Compared with the optimal or normal blood pressure (ONBP), the RR (95%) of CVD deaths with high blood pressure (HBP) were 6.19 (4.85 - 7.91) in male and 2.78 (1.59 - 4.85) in female. In male, compared with ONBP, the RR of CVD deaths without illness but with 1st-grade HBP at baseline, and of those suffered non-CVD but with 1st-grade HBP at baseline, and of those suffered CVD with 2nd-grade HBP at baseline were 3.98, 3.25 and 3.15 respectively (P < 0.01). (3) After stratifying of age, smoking, drinking, educational levels and occupational exposure, the RR of CVD deaths was higher in those who were younger, or ever smoking, non-drinking, higher educational level, exposed to occupational hazards and with hypertension. CONCLUSION: There is relationship between BP levels and CVD mortality. High BP may affect CVD deaths at younger age. Comprehensive measures should be used to reduce the risk of CVD deaths. PMID- 14761570 TI - [Study on the changes in circulating endothelial cells and hemorheology of lung in rats with acute lung injury by chemicals]. AB - OBJECTIVE: To investigate the mechanism of pulmonary circulation obstacle in chemicals-induced acute lung injury and its clinical significance. METHOD: Pulmonary arterial intubation, circulating endothelial cells (CEC) isolation and hemorheology detection technique were used to observe the changes of CEC numbers and hemorheology in rat pulmonary vascular system during oleic acid-induced acute lung injury (ALI). RESULTS: The numbers of CEC were obviously increased even in the early phase of ALI [from (2.06 +/- 0.48)/0.9 micro l to (5.50 +/- 0.54)/0.9 micro l]; there was no obvious change in whole blood viscosity under high shear rate (200 s(-1), 30 s(-1)) but the whole blood viscosity and hematocrit were remarkably increased in pulmonary artery blood at low shear rate (5 s(-1), 1 s( 1)) (P < 0.05). Erythrocytes had increasing tendency, whereas platelets were also decreased but there was no statistical significance. CONCLUSION: The deterioration of pulmonary circulation may be the key point in the pathogenesis of ALI; the injury and dysfunction of pulmonary capillary endothelial cells (PCEC) may be the common starting phase in the pathological processes of ALI; the detection of CEC may offer a new valuable and sensitive index for diagnosis of ALI. PMID- 14761571 TI - [A study on lymphocyte DNA damage in traffic policemen in Guangzhou]. AB - OBJECTIVE: To study the effect of occupational exposure to traffic exhaust and smoking on DNA damage in traffic policemen. METHODS: 812 traffic policemen (741 men and 71 women, 130 of office-work and 682 of outside work) from 8 districts in Guangzhou were investigated. Blood samples were taken by venipuncture and lymphocytes were collected by using lymphocyte separation medium and centrifugation. The comet assay was used to measure DNA damage. RESULTS: The office-work policemen [(37.7 +/- 9.5) years] were older than the outside-work ones [(32.3 +/- 8.1) years, P < 0.001]. No significant difference was observed in sex (P = 0.08) and age (P = 0.45). Comet assay showed that occupational exposure to traffic exhaust significantly increased tail length [4.20 micro m, 95% CI: (3.98 - 4.42) micro m vs 3.23 micro m, 95% CI: (2.82 - 3.7) micro m, P < 0.001]. Smokers had longer tail length [4.66 micro m, 95% CI: (4.37 - 4.97) micro m] than ex-smokers [3.28 micro m, 95% CI: (2.57 - 4.17) micro m] and nonsmokers [3.47 micro m, 95% CI: (3.21 - 3.75) micro m, P < 0.001]. In nonsmokers, significant increase in tail length was observed by passive smoking at home (P = 0.004) but not at work (P = 0.22). When out-door nonsmokers were excluded, passive smoking at work also significantly increased tail length (P = 0.007). Analysis of covariance showed that occupational exposure to traffic exhaust, tobacco smoking, and female had independent effect on lymphocyte DNA damage (P < 0.001) after these factors were adjusted. Passive smoking and age had no effect on lymphocyte DNA damage. CONCLUSIONS: Occupational exposure to traffic exhaust and tobacco smoking respectively increase lymphocyte DNA damage. Female traffic policemen may have more severe DNA damage than male. PMID- 14761573 TI - [Effect of handportable mobiletelephone microwave radiation on rat central neuron apoptosis]. AB - OBJECTIVE: To investigate the effect of handportable mobiletelephone microwave radiation on rat central nervous system by setting up rat model. METHODS: 80 healthy male SD rats (weighed about 200 g) were divided into 4 groups at random: control, radiation, decranium, decranium + radiation. TUNEL method was adopted used to detect the apoptosis of neurons after irradiation, then immunohistochemistry was used to detect Bcl-2, Bax expression in all brain tissue. RESULTS: TUNEL positive rate, Bax and Bcl-2 positive cell numbers could be found in decranium + radiation group [(26.45 +/- 9.27)%, (23.5 +/- 3.58), (11.1 +/- 2.55) respectively]. There were significant differences among control [(9.59 +/- 2.55)%, 14.2 +/- 2.46, 7.0 +/- 1.14 respectively], decranium group [(9.52 +/- 1.93)%, 15.5 +/- 1.77, 7.4 +/- 1.76], radiation group [(10.04 +/- 3.62)%, 15.9 +/- 2.02, 7.2 +/- 1.07] (P < 0.01). But the difference was not found in the ratio of Bax to Bcl-2. CONCLUSION: Microwave radiation did not affect the intact rat, but did promote the occurrence of neuron apoptosis in cranial defect rat. Bax, Bcl-2 gene participated in regulation of apoptosis. The intact cranium may be an important factor to protect the neurons against handportable mobiletelephone microwave radiation to some extent. PMID- 14761574 TI - [Plasma polypeptide hormone levels in rats with gastric ulcer after exposure to intense noise]. AB - OBJECTIVE: To observe changes of plasma polypeptide hormone levels in rats with gastric ulcer after exposure to intense noise, and to discuss their mechanism. METHODS: 80 Wistar rats were used in the study. Plasma levels of rat gastrin (GAS), motilin (MTL), osteocalcin (BGP), substance P (SP), neurotensin (NT) and somatostatin (SS) in rats were measured by radioimmunoassay. RESULTS: (1) In non noise-exposure but with gastric ulcer group, the plasma MTL [(160.70 +/- 40.34) pg/ml] and BGP [(27.63 +/- 13.13) pg/ml] levels on 10 d after gastric ulcer model operation were remarkably higher than those in control group [(89.21 +/- 49.94) pg/ml, (9.10 +/- 1.38) pg/ml respectively] (P < 0.05 and P < 0.01), while the GAS level was remarkably descended [(107.00 +/- 21.75) vs (158.48 +/- 20.92) pg/ml] (P < 0.01). (2) In noise-exposure but without gastric ulcer group, the plasma MTL [(312.80 +/- 207.42) pg/ml] and BGP [(17.76 +/- 12.33) pg/ml] levels on 10 d were also significantly increased as compared with the control group (P < 0.01 and P < 0.05 respectively), while the GAS levels didn't change. (3) In noise-exposure + gastric ulcer group, the areas of gastric ulcer on 10 d and 40 d after noise and operation [(15.33 +/- 7.26) and (15.11 +/- 12.45) mm(2) respectively] were significantly larger than those of the control [(8.22 +/- 6.66), (3.67 +/- 9.90) mm(2)] (P < 0.05). The plasma MTL levels on 10 d and 40 d [(244.44 +/- 68.11) and (191.20 +/- 60.50) pg/ml respectively] were higher than those in control group [(160.70 +/- 40.34) and (93.10 +/- 52.90) pg/ml respectively] (P < 0.01). CONCLUSION: Intense noise exposure may make the rat gastric ulcer worsened and induce negative effect on healing of it. The gastrointestinal endocrine would be disturbed by combined effect of intense noise exposure with gastric ulcer in rats. PMID- 14761575 TI - [Changes of myocardial enzymes in patients with acute carbon monoxide poisoning]. AB - OBJECTIVE: To study the clinical significance of changes of serum myocardial enzymes in patients with acute carbon monoxide poisoning. METHODS: To determine the dynamic changes of the activity of myocardial enzymes and ECG in 62 patients with acute CO poisoning. RESULTS: In patients with acute CO poisoning 5 kinds of myocardial enzymes begin to increase within 24 hours, the activities of aspartate aminotransferase (AST), creatine phosphokinase (CPK), lactic dehydrogenase (LDH), alpha-hydroxybutyrate dehydrogenase (alpha-HBDH), CPK isoenzyme (CK-MB) were (20.2 +/- 12.3), (151.6 +/- 91.8), (146.8 +/- 50.4), (154.8 +/- 47.7), (13.8 +/- 8.1) U/L respectively, while those in control group were (12.1 +/- 6.7), (90.6 +/ 17.3), (118.7 +/- 13.5), (89.9 +/- 27.9), (5.9 +/- 3.3) U/L respectively. There was significant difference between two groups (P < 0.01); 3 d later, the activities of 5 enzymes were still increased [(21.3 +/- 12.3), (105.8 +/- 51.4), (144.8 +/- 51.4), (159.8 +/- 35.4), (16.2 +/- 9.1) U/L respectively]. 7 and 12 d later, the activities of alpha-HBDH and CK-MB were still higher than those of control (P < 0.01). LDH(1) and LDH(2) increased to peak value in 24 h after poisoning (35.3 +/- 5.8), (43.8 +/- 5.7) U/L vs (24.8 +/- 3.9), (36.9 +/- 4.3) U/L, P < 0.01. The abnormal rate of serum LDH(1) was 78.7%, LDH(2) 58.3%, LDH 45.2%, CK-MB 37.1%, alpha-HBDH 33.6% and the abnormal rate of ECG was less than 10%. CONCLUSION: Acute carbon monoxide poisoning may cause myocardial injury. Determination of serum myocardial enzymes may contribute to showing myocardial injury, early diagnosis and treatment, results of treatment and prognosis. PMID- 14761576 TI - [The effect of local vibration on blood-lipids and whole blood viscosity]. AB - OBJECTIVE: To study the effects of local vibration on blood-lipids and whole blood viscosity. METHODS: The total cholesterol (TC), triglyceride (TG), high density lipoprotein (HDL), low density lipoprotein (LDL), whole blood viscosity, apolipoprotein (Apo-), red blood cell (RBC), platelet (PLT), mean corpuscular volume (MCV), serum-protein, postprandial blood sugar (PBS), and serum-protein of experimental and control workers were detected. The difference of the means and abnormal rates of two groups were compared. RESULTS: The means of TG, TC, HDL in exposed group [(1.01 +/- 0.85), (3.25 +/- 0.61), (1.14 +/- 0.20) mmol/L respectively] were significantly lower than that of control group [(1.89 +/- 1.47), (3.87 +/- 0.82), (1.22 +/- 0.26) mmol/L, respectively, P < 0.01 or P < 0.05]. Apo-A was also decreased [(1.13 +/- 0.29) g/L vs (1.23 +/- 0.16) g/L, P < 0.01]. The mean of whole blood viscosity were significantly increased in exposed group [(2.76 +/- 0.42) mPa.s vs (2.54 +/- 0.33) mPa.s, P < 0.01]. The abnormal rate of Apo-A was significantly higher in exposed group (23.30%) than that in control (4.50%, P < 0.01). CONCLUSION: Local vibration may induce decrease in blood lipids, increase in blood viscosity and changes in some other blood parameters. PMID- 14761578 TI - [Construction of eukaryotic expression vector of hMTH1 gene antisense RNA]. AB - OBJECTIVE: To construct pEGFP-C1-T vector, an eukaryotic expression plasmid of hMTH1 gene antisense RNA. METHODS: The conservative region of hMTH1 gene was amplified by RT-PCR after total RNA being extracted from human embryo lung fibroblast (HLF) and then cloned into pGEM-T vector. After the recombinant plasmid was certified by DNA sequencing, the conservative region of hMTH1 gene was inserted into pEGFP-C1 vector reversedly and pEGFP-C1-T vector was constructed. The efficiency of antisense inhibition was verified by Western blotting after cell transfection. RESULTS: 423 bp fragment including conservative region of hMTH1 gene was obtained by RT-PCR. After cloned by pGEM-T vector and certified by DNA sequencing, pEGFP-C1-T vector was successfully constructed by means of recombinant DNA technology. Additionally pEGFP-C1-T vector could efficiently decrease hMTH1 protein level by 46%. CONCLUSION: The efficient expression vector of hMTH1 gene antisense RNA, pEGFP-C1-T has been constructed successfully. PMID- 14761593 TI - [Pathological features of light chain nephropathy]. AB - OBJECTIVE: To investigate the pathologic features and diagnostic algorithm of light chain nephropathy (LCN). METHODS: Seven cases of LCN were studied by light microscopy, electron microscopy and immunolabeling of light chains (kappa, lambda) by immunofluorescence and immunoelectron microscopy. RESULTS: The histopathology of 7 cases by light microscopy was variable, with 3 cases showing nodular glomerulosclerosis, 1 case showing mild to moderate mesangial proliferation, and 3 cases showing cast nephropathy with minimal glomerular change. Immunofluorescence study revealed positive staining of a single type of light chain in mesangium (nodular pattern) or along glomerular basement membrane (linear), along tubular basement membrane and around arteriolar walls in all the 7 cases. Ultrastructurally, electron-dense granular deposits were identified in mesangium, subendothelial aspect of glomerular basement membrane, outer aspect of tubular basement membrane and arteriolar walls. Immunogold labeling of light chains showed distinct labeling of a single type light chain in the granular electron-dense materials (5 cases being kappa-positive and 2 being lambda positive). CONCLUSIONS: LCN typically shows nodular glomerulosclerosis. The ultrastructural change is characteristic and important for diagnosis. Immunolabeling of light chains by immunofluorescence and immunoelectron microscopy carries further diagnostic value, especially in cases with minimal light microscopic change. PMID- 14761594 TI - [Juxtaglomerular cell tumor of the kidney: a clinicopathologic analysis of five cases]. AB - OBJECTIVE: To study the morphologic characteristics and immunophenotype of juxtaglomerular cell tumor of the kidney (JGCT), with discussion on its diagnostic clues and possible histogenesis. METHODS: The clinical, pathologic and immunohistochemical features of 5 cases of JGCT were evaluated. In addition, 5 cases of hemangiopericytoma and 5 cases of cutaneous glomus tumor were selected for comparative immunohistochemical analysis. RESULTS: The JGCT cases came from 4 females and 1 male (mean age at diagnosis = 32 years). All of them manifested symptoms of systemic hypertension. Four of the patients received partial nephrectomy and the remaining patient was treated by radial nephrectomy. All of them were followed up for a period of 4 to 66 months (average = 27 months). There was no evidence of local recurrence or distant metastases. On gross examination, these JGCTs were well-circumscribed and situated in the renal cortex and measured 4.4 cm in greatest dimension on average. Histologically, the tumor was characterized by the following three features: (1) solid sheets of relatively uniform polygonal to round cells with lightly eosinophilic cytoplasm, sometimes containing PAS-positive intracytoplasmic granules; (2) absence of or very scanty mitotic figures; (3) interstitium rich in thin-walled capillaries, associated with focal hyaline change and hemangiopericytoma-like architectural pattern. Under electron microscopy, characteristic rhomboid-shaped renin granules were found in the cytoplasm. All JGCTs were immunoreactive for renin, CD34, actin, and calponin. In contrast, all glomus tumors were negative for renin and all hemangiopericytomas were negative for actin. CONCLUSIONS: JGCT is a rare benign renal neoplasm typically found in young adults and manifests as systemic hypertension. The tumor cells may be originated from modified vascular smooth muscle cells. The identification of renin granules by electron microscopy and demonstration of the characteristic immunophenotype is the key to correct pathologic diagnosis. PMID- 14761595 TI - [Morphological study of severe acute respiratory syndrome (SARS)]. AB - OBJECTIVE: Seven cases of autopsy from SARS patients are studied to investigate the pathogenesis and the pathologic changes of the major organs. METHODS: Detailed gross and microscopic examination of the autopsy specimen is performed, including lung, heart, liver, kidney, spleen and lymph nodes. RESULTS: All of the lungs are markedly enlarged and consolidated. Microscopically, pulmonary edema is a prominent finding, especially at the early stage of the disease (5 days after the onset). The alveolar spaces are filled with fibrinous exudates and lined with hyaline membrane. In 5 cases that undergo over 3 weeks of the course, the main pattern is organization of intra-alveolar deposit, along with fibroblastic proliferation in the alveolar septa, which leads to obliteration of alveolar space and pulmonary fibrosis. All of the lungs show bronchopneumonia, scattered hemorrhage, and proliferation of alveolar epithelial cells with desquamation. Microthrombi are seen in 6 cases. Fungal infection is noted in 2 cases. One of them is disseminative, involving bilateral lungs, heart, and kidney; the other one is diagnosed in hilar lymph nodes. In immune system, hilar and abdominal lymph nodes are usually congested and hemorrhagic, with depletion of lymphocytes, and accompanied with subcapsular sinus histiocytosis. One of the cases shows enlargement of abdominal lymph nodes, which have reduced number of germinal centers. Spleen exhibits atrophy of white pulps, and even lost of white pulps in some areas. The red pulp is markedly congested and hemorrhagic. In 5 cases, cardiomegale is prominent. Thrombosis (2 cases), focal myocarditis (1 case), and fungal myocarditis (1 case) are observed. In addition, liver shows massive necrosis (1 case) and nodular cirrhosis (1 case). CONCLUSIONS: Lung is the major organ affected by SARS, demonstrated as diffuse alveolar damage. It is postulated that viral infection induces severe damage of alveolar epithelial and capillary endothelial cells, leads to pulmonary edema, intra-alveolar fibrin deposit, and hyaline membrane formation. Consequently, intra-alveolar organization and alveolar septal fibrosis causes loss of alveolar spaces, eventually, pulmonary fibrosis and atelectasis. The immune system is often affected, and presented as depletion of lymphoid tissue in lymph nodes and spleen. Secondary infection is a common complication, which should be paid close attention in the management of SARS patients. PMID- 14761596 TI - [A clinicopathological study of Castleman's disease]. AB - OBJECTIVE: To study the pathogenesis and the differential diagnosis of Castleman's disease. METHODS: Histopathology, immunohistochemical staining and clinical courses of 26 cases of Castleman's disease (CD) were studied with follow up study of 16 cases. RESULTS: The present study included 6 cases of multicentric type, 20 cases of localized type in the clinical aspects and 19 cases with hyaline vascular type, 4 cases with plasma cell type, 3 cases with mixed type in the histologic aspect. The Multicentric type presented systemic lymphadenopathy, anemia, hyperglobulinemia, hepatosplenomegaly, skin changes, and lung disorder and kidney disfunction, of which 1 case died of respiratory and renal insufficiency. 13 of the 20 localized cases were of the hyaline vascular type, and with good prognosis. 7 of the 20 cases showed paraneoplastic pemphigus associated with hyperglobulinemia (4/7) and lung disease (5/7). The pathologic features composed of proliferation of the mantle zone B cell, follicular dendritic cell, plasma cell and small vessels. In immunohistochemical staining, kappa and lambda light chains were detected in each CD case. CONCLUSIONS: Many diseases are similar to CD clinicopathologically. It is important to make differential diagnosis through pathological study. Castleman's disease is a lymphoproliferative disorder. The pathogenesis of this multicentric disorder may be associated with autoimmune disease. PMID- 14761597 TI - [The diagnosis and differential diagnosis of histiocytic necrotizing lymphadenitis. A study of histology, immunohistochemistry and electron microscopy]. AB - OBJECTIVE: To study the diagnosis and differential diagnosis of histiocytic necrotizing lymphadenitis (Kikuchi disease, KD). METHODS: Histologic analysis and immunohistochemical study (EnVision method) was carried out in 46 cases of KD, 5 cases of nonspecific lymphadenitis (NLD), 5 cases of non-Hodgkin's lymphoma (NHL), 5 cases of Hodgkin's lymphoma (HD), 5 cases of cat-scratch disease (CSD) and 5 cases of tuberculous lymphadenitis (TBL). Electron microscopy was also performed in 6 cases of KD and 2 cases of NHL. RESULTS: Three histologic (proliferative, necrotizing and xanthomatous) patterns were noted in KD. In any of these patterns, there were some basic histologic findings: a wedge-shaped pale area at the edge of the lymph node or paracortical nodules associated with an increase in apoptotic cells or karyorrhectic debris, crescentic histiocytes, proliferative mononuclear histiocytes and absence of or very scanty neutrophils. Immunohistochemical study demonstrated focal occurrence of histiocytes expressing both CD68 and MPO. Electron microscopy confirmed the presence of apoptotic bodies, proliferative mononuclear histiocytes, crescentic histiocytes and dispersed T cells in the lesional areas. CONCLUSIONS: In general, there should not be much difficulty in differentiating KD from other types of lymphadenopathy. Sometimes, problems arise mainly because of the diversity of KD histology. Correct diagnosis can be made if one pays attention to the described characteristic morphology, peculiar immunophenotype of the histiocytes and possibly ultrastructural features. PMID- 14761598 TI - [Expression of cytokeratins and ret in thyroid papillary carcinoma]. AB - OBJECTIVE: To investigate the expression of cytokeratins and ret in thyroid papillary carcinoma (TPC) and their diagnostic value. METHODS: During the period of October 1999 to March 2002, 69 cases of TPC (42 cases with adjacent normal thyroid tissue) and 14 cases of nodular goiter with papillary hyperplasia were enrolled into the study. Immunohistochemistry for CK19, CK17, CK8, CK20 and ret was performed in all cases using EnVision and LSAB methods respectively. RESULTS: The positive rates for CK19 and ret in TPCs were 85.5% and 68.1% respectively, which were significantly (P < 0.01) higher than those in nodular goiter and normal thyroid tissue (25.0% and 5.4% respectively). The expression of CK17 was also observed in a few cases of TPCs (11/69, 15.9%), which was mainly localized in areas of squamous metaplasia, poorly differentiated carcinoma and/or in the small infiltrative foci. The positive rates for CK8 were 75.4% and 26.8% in TPCs and benign thyroid tissue respectively. All cases were negative for CK20. CONCLUSIONS: CK19, CK17 and ret expressions are significantly higher in TPCs than benign thyroid tissue; and this characteristic can have important diagnostic value. PMID- 14761599 TI - [Analysis of Epstein-Barr virus with BamHI "f" variant and XhoI-loss of LMP1 gene in nasopharyngeal carcinoma]. AB - OBJECTIVE: To investigate the genomic variation of Epstein-Barr virus (EBV) and its significance in nasopharyngeal carcinogenesis. METHODS: Forty nasopharyngeal carcinoma (NPC) biopsy tissues were used for detection of EBV BamHI f variant and LMP1 XhoI-loss by polymerase chain reaction (PCR), nested PCR, and RFLP (restriction fragment length polymorphism). Forty-eight samples of peripheral blood mononuclear cells (PBMC) taken from apparently healthy adult individuals were used for detection of LMP1 XhoI-loss. Three samples of amplified LMP1 exon 1 DNA from B95-8 cell line and 2 NPC tissues (one having XhoI-loss and the other having Wt-XhoI/XhoI-loss) were sequenced. RESULTS: Thirty out of the 40 NPC cases (30/40, 75%) harbored EBV BamHI f variant and the remaining 10 (10/40, 25%) harbored BamHI F prototype. Thirty out of the 39 NPCs (30/39, 76.9%) showed single EBV LMP1 XhoI-loss, 7 (7/39, 18.0%) showed single LMP1 Wt-XhoI (presence of a XhoI site in exon 1 of LMP1 gene, as in B95-8 cell line), and 2 (2/39, 5.1%) showed both LMP1 Wt-XhoI and XhoI-loss. Thirty-eight of the 39 NPCs (97.4%) showed EBV LMP1 XhoI-loss or/and BamHI F variant. In the NPC tissue (1 case only) showing the prototype of Wt-XhoI/BamHI "f", there were several base substitutions, including 5 missense mutations and 2 silent mutations present in LMP1 exon 3, on DNA sequencing. On the other hand, 10 out of the 48 samples of PBMC taken from apparently healthy individuals could be amplified successfully by nested PCR for detection of LMP1 XhoI site. All of these 10 samples carried the prototype of EBV LMP1 Wt-XhoI. CONCLUSIONS: The majority of EBV present in neoplastic cells of NPC is of BamHI "f" variant and/or possesses LMP1 XhoI-loss, as compared with that in healthy individuals. This genomic variation of EBV may bear some roles in the development and progression of NPC. PMID- 14761600 TI - [Study on vasculogenic mimicry in malignant melanoma]. AB - OBJECTIVE: To investigate the mode of angiogenesis between highly invasive malignant melanoma and poorly invasive malignant melanoma by immunohistochemistry and periodic acid-Schiff stain (PAS) and to discuss whether the tumor cells in highly invasive malignant melanoma carry vasculogenic mimicry through self metamorphosis, thus acquiring blood supply to sustain their growth. METHODS: Thirty cases of highly invasive malignant melanoma and 30 cases of poorly invasive malignant melanoma were retrieved and reprocessed as tissue microarray for further investigations. The tissue microarray sections were then stained with CD34 and PAS; and the positivity rates were compared. RESULTS: There was a significant difference between CD34 and PAS staining in highly invasive malignant melanoma (P < 0.01). The difference was not statistically significant in poorly invasive malignant melanoma (P > 0.05). CONCLUSION: Vasculogenic mimicry exists in some cases of highly invasive malignant melanoma. It is possible that the tumor cells can acquire blood supply to sustain growth and metastasize via this mechanism. PMID- 14761601 TI - [Changes of matrix metalloproteinase-2 and tissue inhibitor of metalloproteinase 2 expressions on cultured rat mesangial cells transfected with Smad 7 vector]. AB - OBJECTIVE: To investigate matrix metalloproteinase-2 (MMP-2) and tissue inhibitor of metalloproteinase-2 (TIMP-2) expressions in the cultured rat mesangial cells (MsC) transfected with Smad 7 vector and to elucidate the mechanism of Smad 7 in blocking tissue fibrosis. METHODS: Lipofectin method was used to transfect Smad 7 vector into MsC. Western blot and RT-PCR analyses were then used to detect Smad 7 protein and mRNA expression levels. The expressions of MMP-2 and TIMP-2 were determined by Western blot, RT-PCR and zymography assay. RESULTS: Two MsC clones (S-22, S-26) with Smad 7 overexpression were successfully established. The two clones showed an increased expression of MMP-2 protein and enhanced enzyme activity. The expressions of TIMP-2 protein and mRNA however were suppressed. CONCLUSIONS: It is possible that Smad 7 can alleviate the development of tissue fibrosis by upregulating the expression of MMP-2 and downregulating the expression of TIMP-2 in mesangial cells. PMID- 14761602 TI - [Expression of monocyte chemoattractant protein-1 in experimental rat glomerulonephritis is mediated by NF-kappaB/IkappaB signal pathway]. AB - OBJECTIVE: To investigate the role of NF-kappaB/IkappaB signal pathway in mediating the expression of monocyte chemoattractant protein-1 (MCP-1) in experimental rat glomerulonephritis. METHODS: Nephrotoxic serum nephritis (NTN) was induced by injection of anti-GBM antibody into the tail veins of rats. Electrophoretic mobility shift assay (EMSA) and Western Blot were used to detect the activation of NF-kappaB, nuclear translocation of p65 subunit and degradation of IkappaBalpha and IkappaBbeta in rat renal tissue. MCP-1 expression in glomeruli and renal tubules was also assessed by immunohistochemistry and ribonuclease protection assay. This was further correlated with the activation of NF-kappaB. RESULTS: There was an obvious expression of MCP-1 in glomeruli and renal tubules. Significant up-regulation of NF-kappaB activation, nuclear translocation of p65 subunit, and degradation of IkappaBalpha and IkappaBbeta were also observed in NTN rat renal tissue, as compared to the control group. A positive correlation was noted between NF-kappaB activation and MCP-1 expression. CONCLUSIONS: NF-kappaB/IkappaB signal pathway may play an important pathogenetic role in glomerulonephritis, with mediating the expression of MCP-1. PMID- 14761603 TI - [Effect of transforming growth factor beta1/Smad signaling pathway on the expression and enzymatic activity of matrix metalloproteinase-2 and tissue inhibitor of metalloproteinase-2 in cultured rat mesangial cells]. AB - OBJECTIVE: To explore the effect of transforming growth factor (TGF) beta1/Smad signaling pathway on the expression and enzymatic activity of matrix metalloproteinase-2 (MMP-2) and tissue inhibitor of metalloproteinase-2 (TIMP-2) in cultured rat mesangial cells (MsC). METHODS: Lipofectin method was used to transfect Smad 2, Smad 3 and Smad 7 vectors into MsC; and immunofluorescence, RT PCR and Western blot analysis were used to detect their transfection efficiency. The expression and enzymatic activity of MMP-2 and TIMP-2 were determined by Western blot, zymography or reverse zymography assay. RESULTS: MsC transfected with Smad 2 gene showed slightly increased expression and enzymatic activity of both MMP-2 and TIMP-2, which was more obvious upon stimulation by TGF-beta1. MsC transfected with Smad 3 gene showed a slight upregulation of TIMP-2 expression and its enzymatic activity, which was enhanced after TGF-beta1 stimulation. There was however no change in MMP-2 expression and its enzymatic activity. On the other hand, MsC transfected with Smad 7 gene showed a decrease in MMP-2 and TIMP 2 expression and enzymatic activity, which was especially obvious after stimulation by TGF-beta1. CONCLUSIONS: TGF-beta1/Smad signaling pathway may play an important role in the pathogenesis of glomerulosclerosis, probably via MMP-2 and TIMP-2 expression and the associated enzymatic activity. PMID- 14761605 TI - [Reversal of multidrug resistance property of carcinoma cells by down-regulating transcription of mdr-1]. AB - OBJECTIVE: To reverse the multidrug resistance (MDR) property of carcinoma cells by blocking transcription of activating sites of mdr-1. METHODS: Breast carcinoma cells were transinfected with several antisense oligonucleotide (ASODN) complementary to mdr-1 by lipofectin. RT-PCR was used to detect the production of mdr-1mRNA. The expression of P-glycoprotein (gp) was then detected by immunohistochemistry and the function of P-gp was detected by rhodamine123 retention. RESULTS: Forty-eight hours after transfection, mdr-1 index of cells treated by ASODN complementary to MA zone (major initiation start zone), MI (minor initiation start zone), C zone (CAAT box), G zone (GC box) of mdr-1 gene was 1.4, 1.9, 1.6 and 2.1 respectively. The rate of P-gp protein expression in treated cells was 14%, 43%, 26% and 39% respectively. The intracellular Rh123 retention in treated cells was 125%, 83%, 102% and 77% respectively. There was significant difference between cells treated by ASODN complementary to MA zone and C zone and drug-resistant cells. CONCLUSIONS: The ASODN complementary to MA zone and C zone of mdr-1 gene can reverse MDR of drug-resistant cells to various extent, amongst which the former is more effective. Down-regulating transcription of mdr-1 by blocking transcription activating sites can reduce the expression of mdr-1mRNA and P-gp, and thus reversing MDR of carcinoma cells. The ASODN complementary to MI zone, G zone of mdr-1 however do not significantly reverse the MDR property. PMID- 14761604 TI - [Expression of early growth response gene-1 in macrophages stimulated by silicon dioxide]. AB - OBJECTIVE: To study the expression and localization of early growth response gene 1 (Egr-1) in macrophages after stimulation by silicon dioxide in vivo and in vitro and to discuss the role of Egr-1 in the development of silicosis. METHODS: The expression of Egr-1 in animal model of silicosis was analyzed by using immunohistochemistry. Western-blot, immunofluorescence and RT-PCR analysis were used to detect the expression and localization of Egr-1 protein and the dynamic changes of Egr-1 mRNA in cultured macrophages RAW264.7, after stimulation by silicon dioxide. RESULTS: In animal model with induced silicosis, there was an increased expression of Egr-1 in pulmonary macrophages. The expression levels peaked at the 14th day. In vitro, the transcription of Egr-1 increased in RAW264.7 macrophages during 15 to 240 minutes after the administration of silicon dioxide. The response peaked at 15 minutes and diminished to a minimal level at 480 minutes. Nuclear translocation was most apparent at 60 minutes, lasted till 120 minutes and diminished gradually. During the period from 60 to 120 minutes, the expression of Egr-1 protein also reached a peak. CONCLUSIONS: Silicon dioxide can activate the nuclear transcription factor Egr-1 in vivo and in vitro in macrophages. Egr-1 may thus play an important pathogenetic role in the development of silicosis. PMID- 14761606 TI - [CIK cells acquired multidrug resistance and maintained cytotoxic activity to tumor cells after mdr1 gene transfection]. AB - OBJECTIVE: To investigate whether the cytokine-induced killer (CIK) cells could acquire multidrug resistance and maintain the original cytotoxic activity after multidrug resistance (mdr1) genes transfection. METHODS: CIK cells were generated from peripheral blood cultured with IFN-gamma, CD(3) monoclonal antibody, IL-2, IL-1 and transfected with a plasmid (pHamdr) containing human mdr1 gene via electroporation. RT-PCR method was used to assay mRNA expression of mdr1 gene in transfected CIK cells, flow cytometry with anti-P-gp monoclonal antibody to detect P-glycoprotein (P-gp) expression on CIK cells membrane, and MTT assay to compare both the multidrug resistance to doxorubicin and colchicines and cytotoxic activity to human mammary cancer cell line MCF7 between transfected and non-transfected CIK cells. RESULTS: mdr1 expression was detected in the transfected CIK cells. There was a strong expression of P-gp on the transfected CIK cells and the percentages of P-gp positive cells were 21% - 37% (average 27%). The IC(50) of transfected CIK cells to doxorubicin was 22.3 - 45.8 times and 6.7 - 11.35 times to colchicines of those of non-transfected CIK cells. The cytotoxic activity to MCF7 remained unchanged (P > 0.05). CONCLUSION: It demonstrated that CIK cells transfected with mdr1 gene via electroporation could express multidrug resistance successfully without changes of cytotoxic activity. PMID- 14761607 TI - [Topoisomerase II inhibitors etoposide and doxorubicin induced rearrangement and fusion of AML1 gene]. AB - OBJECTIVE: To investigate the relationship between topoisomerase II inhibitors and t(8;21) chromosomal translocation. METHODS: The rearrangements of AML1 and ETO genes were detected by Southern Blot and the AML1-ETO fusion gene by nested RT-PCR combined with sequencing in K562 cells treated with etoposide (Vp16) and doxorubicin (DOX). RESULTS: The rearrangements of AML1 gene were detectable after DOX treatment at concentrations of 10, 50 and 100 micro mol/L for 16 h, AML1-ETO fusion gene appeared after 50 micro mol/L DOX treatment for 48 h. CONCLUSION: Induction of AML1 gene rearrangement and fusion by topoisomerase II inhibitors, represents one of the molecular mechanisms of t(8;21) chromosomal translocation. PMID- 14761608 TI - [Comparative proteomics research of apoptosis initiation induced by homoharringtonine in HL-60 cells]. AB - OBJECTIVE: To study the related proteins of apoptosis initiation induced by homoharringtonine (HHT) in HL-60 cells. METHODS: After establishment of an apoptosis initiation model induced by HHT in HL-60 cells, proteins of untreated and HHT treated HL-60 cells were extracted, and the two-dimensional polyacrylamide gel electrophoresis (2-DE) maps of the extracted proteins were established by using the immobilized pH gradient (IPG) two-dimensional electrophoresis respectively. The alteration protein spots were identified with assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) and database searching. RESULTS: Proteomics analysis showed that proteins including MHC class I antigen, calbindin D-28K, chloride channel protein 6, oncoprotein 18, zinc finger protein Helios and apoptosis inhibitor like protein 2 were involved in apoptosis initiation induced by HHT. CONCLUSION: The present study might conduce to the researches of HL-60 cells carcinogenesis and pave the way to exploit drug precursor related to HHT and initiation of apoptosis in HL-60 cells. PMID- 14761609 TI - [Indomethacin-induced HL-60 cell apoptosis is associated with inhibition of beta catenin/c-myc signal transduction pathway]. AB - OBJECTIVE: To investigate the effects of indomethacin on HL-60 cell proliferation and apoptosis, and elucidate partly the molecular mechanism about the anti leukemia effect of indomethacin by studying beta-catenin signal transduction pathway. METHODS: HL-60 cells were treated with indomethacin at different concentrations (0, 25, 50, 100, 200, 400 micro mol/L). Cells viability and proliferation were determined by Trypan blue staining and cell counting respectively. DNA ladder pattern and cell morphology were used to identify cell apoptosis. The expression and cleavage of caspase-3, beta-catenin, c-myc were detected by Western blot techniques. RESULTS: Indomethacin could significantly inhibit HL-60 cells proliferation and induce cells apoptosis with a time and dose dependent manner. An up-regulating expression and cleavage of caspase-3 were observed. Indomethacin could inhibit beta-catenin expression and induce its degradation, c-myc protein exhibited a down-regulation with a concentration dependent manner. CONCLUSION: Indomethacin could inhibit HL-60 cell proliferation and induce cells apoptosis. Anti-leukemia effect of indomethacin was associated with the inhibition of "beta-catenin --> c-myc" signal pathway. PMID- 14761610 TI - [Experimental study of the specific immunoresponse induced by p53 modified dendritic cells]. AB - OBJECTIVE: To explore the specific cytotoxic T lymphocyte (CTL) induced by dendritic cells (DC), which were transfected by the plasmid pC53-SN3 encoding p53 gene. METHODS: DC derived from HLA-A2(+) mononuclear cells of the 24-lung cancer patients was transfected with the plasmid pC53-SN3 by lipofectamine and then co cultured with auto-unpurified T cells to induce potent CTL (T-pC53-SN3). The cytolysis of specific CTL against Calu-6, a HLA-A2(+) human lung cancer cell line, was measured by using lactate dehydrogenase (LDH) releasing assay. RESULTS: The expression of CD(1a) and CD(83), the correlative markers of DC, increased apparently after transfected with plasmid pC53-SN3, the expression rate was (5.45 +/- 0.89)% and (3.26 +/- 0.47)% versus (52.15 +/- 11.56)% and (25.78 +/- 12.35)%. CD(14) decreased apparently, but other DC correlative markers of CD(1a), CD(40), CD(86), and HLA-DR remained almost the same as that before transfection. Compared with T-IL-2, the CTL derived from PBMNC stimulated by IL-2 (100 U/ml), the cytolytic activity of T-pC53-SN3 against Calu-6 cell line showed a significant increase, but cytolytic activity was 56.79 +/- 15.67 and 39.33 +/- 9.88, respectively, when effect cells: target cells was 10:1. The expression of the CD(8), CD(69), and CD(45)RO/CD(8) of T-pC53-SN3 cells increased significantly, but that of CD(3), CD(4), CD(86), ect, was not significantly different from those of T-pCMV-neo. CONCLUSIONS: It showed that DC transfected by p53 gene could induce potent HLA-A(2) restrictive CTL to kill tumor cell efficiently. PMID- 14761611 TI - [Inhibition effect of CIITA anti-sense cDNA on the expression of MHCII molecules of Hela cells]. AB - OBJECTIVE: To investigate the inhibiting effect of classII major histocompatibility complex transactivator (CIITA) anti-sense cDNA on MHCII molecules expression. METHODS: CIITA antisense cDNAs (arII1, arII2 and arII3) were obtained from Raji cell by RT-PCR, and then inserted into the pcDNA3.1B plasmid. The stable transfectants of Hela cell were tested for CIITA protein by immunohistochemistry, and classic MHCII molecules (HLA-DR, -DP, -DQ) induction through recombinant human IFN-gamma by flow cytometry. mRNAs of CIITA, invariant chain and classic MHCII molecules were detected by RT-PCR. RESULTS: The effect of arII2 was the best. When induced with IFN-gamma, the expression of CIITA protein was inhibited by 87.23%, and that of HLA-DR, -DP and -DQ was almost totally inhibited on arII2 positive Hela cells. The mRNA contents of CIITA, HLA-DR, -DP, DQ and invariant chain were decreased significantly (P < 0.05). CONCLUSIONS: arII2 inhibits CIITA and thus the family of MHCIImolecules regulated by CIITA, therefore, it provides a novel approach to graft versus host disease study in hematopoietic stem cell transplantation. PMID- 14761612 TI - [New insight into the mechanism of p53 inducing leukemia cell apoptosis]. AB - OBJECTIVE: To investigate the expression changes of intrinsic cytokines TGF beta(1) and TNF-alpha, telomerase activity and bcl-2 during ongoing apoptosis of HL-60 and K562 cells induced by p53. METHODS: pN53cG (Val135), a temperature sensitive p53 mutant, which behaved like wild type p53 (wt-p53) at 32.5 degrees C, were introduced into p53-null HL-60 and K562 cells respectively by lipofectin. In the presence of G418, HL-60-pN53cG and K562 pN53cG clones expressing p53 protein were selected. The ongoing expression of intrinsic cytokines (TGF-beta(1) and TNF-alpha), bcl-2 oncogene and hTERT mRNA during the apoptosis of HL-60 and K562 cells induced by p53 and the effects of exogenous p53 gene, TGF-beta(1) and TNF-alpha antisense PS-ODNS on the apoptosis of HL-60 and K562 cells and the expression of bcl-2 were studied by RT-PCR, quantitative RT-PCR, DNA fragmentation, TdT-mediated dUTP nick end labeling (TUNEL) and flow cytometery. The levels of secreted TGF-beta(1) and telomerase activity were detected by ELISA and PCR-ELISA, respectively. RESULTS: (1) The expressions of intrinsic TGF beta(1) and TNF-alpha mRNA were up-regulated, while that of bcl-2 and hTERT down regulated. The levels of TGF-beta(1) in the supernatant of HL-60 and K562 cells were increased, and the level of telomerase activity decreased. (2) Antisense PS ODNS of TGF-beta(1) and TNF-alpha could obviously inhibit the p53 inducing cell apoptosis, and restore bcl-2 mRNA and protein to pre-treated level. CONCLUSIONS: Exogenous p53 induces leukemia cell apoptosis via up-regulating the expression of intrinsic TGF-beta(1) and TNF-alpha and down-regulating the expression of hTERT and bcl-2. PMID- 14761613 TI - [Activating effects of protein transduction domain mediated BCR/ABL protein on CML T cells]. AB - OBJECTIVE: To study the activating effect of protein transduction domain (PTD) mediated BCR/ABL protein on T cells from CML patients. METHODS: The plasmid containing PTD and b3a2 bcr/abl of CML was constructed by genetic engineering and expressed in E. coli. The peripheral blood mononuclear cells from CML patients were stimulated in vitro with purified PTD-BCR/ABL protein and the expression of the early activation antigen CD(69) on CD(8)(+) and CD(4)(+) T cells was detected by flow cytometry (FCM). RESULTS: The optimal concentration of PTD-BCR/ABL protein for activating CD(8)(+) T cells in vitro was 100 micro g/ml, CD(69) expression peaked in three days stimulation. CD(8)(+) T cells were activated in 10 of 15 CML patients, the expression rate of CD(69) was (15.01 +/- 3.75)%. CD(4)(+) T cells were activated in 4 of 15 patients, the expression rate of CD(69) was (10.32 +/- 3.08)%. Both CD(8)(+) and CD(4)(+) T cells were activated simultaneously in 3 of them. However, neither CD(4)(+) nor CD(8)(+) T cells was activated by stimulation with BCR/ABL protein in all 15 specimens, the expression rate of CD(69) on CD(8)(+) and CD(4)(+) T cells was (1.36 +/- 0.31)% and (1.41 +/ 0.43)%, respectively. There was no difference compared with that of PBS control group (P > 0.05). CONCLUSION: By using a PTD-mediated antigen delivering system, exogenous BCR/ABL protein can be delivered into APC, processed and presented onto surface of APC to activate Ag-specific CD(8)(+) and CD(4)(+) T cells in vitro. PMID- 14761614 TI - [Effect of cyclin G1 antisense oligodeoxynucleotides (ASON) on the growth of HL 60 cells in nude mice]. AB - OBJECTIVE: To study the inhibition effect of cyclin G(1) antisense oligodeoxynucleotides (ASON) on the growth of HL-60 cells in nude mice. METHODS: (1) Nude mice were divided into control group, sense oligodeoxynucleotides (SON) group and ASON group. After (60)Co radiation, with HL-60 cells SON group and ASON group were subcutaneously innoculated; (2) The weight and volume of tumors were continually measured; (3) The morphology of tumor cells was observed by microscope; (4) The protein and mRNA expression levels of cyclin G(1) were determined by flow cytometry (FCM) and reverse transcription polymerase chain reaction (RT-PCR); (5) The cell apoptosis was detected by electron microscopy and FCM. RESULTS: (1) The inhibition rate of tumor in ASON group was 69.4%. In ASON group, the wight and volume of tumor were significantly lower than those in SON group and control group. (2) The HL-60 cells in ASON group showed morphologically smaller nuclei, less mitosis, less heteromorphosis and apoptosis. CONCLUSION: The cyclin G(1) ASON can inhibit the growth of HL-60 cells in nude mice and induce apoptosis. PMID- 14761615 TI - [Isolation and characterization of human placenta derived adherent cells and their hematopoietic growth factor expression]. AB - OBJECTIVE: To isolate and culture human placenta derived adherent cells (hPDAC) and assay their hematopoietic growth factor expression. METHODS: By enzyme digestion, hPDAC were isolated from human placenta tissue and cultured, and their biological characteristics were studied. The hematopoietic growth factor (HGF) mRNA expression of hPDAC was assayed by RT-PCR. RESULTS: hPDAC was successfully isolated from human placenta tissue, which was further confirmed as mesenchymal stem cell-like cells. HGF including SCF, FL, G-CSF, GM-CSF, M-CSF and IL-6 were expressed in hPDAC. CONCLUSION: hPDAC could be used as feeder layer for umbilical cord blood CD(34)(+) cells ex vivo expansion. PMID- 14761616 TI - [A cross-sectional study on the usage condition of hypertension related examinations and antihypertensive drugs among 40 district and community based hospitals in Beijing]. AB - OBJECTIVE: To study the use of hypertension related physical examinations, laboratory tests and anti-hypertension drugs among district and community based hospitals in Beijing and to analyze the possible hardware (examination, test, drug) problems under the application of the "China Hypertension Guide" in those hospitals. METHODS: The survey was carried out in 40 hospitals in Chaoyang and Haidian districts, including 9 district level hospitals, and the rest at community level. Questionnaire survey and in-depth interview were used to collect information from the leaders of related departments in those hospitals. RESULTS: (1) The hardware condition of district hospital satisfied the implicit requirements of hypertension evaluation and treatment in "China hypertension Guide". (2) 64.5% of the community level hospitals had the basic equipments and routine laboratory tests for diagnosis and treatment on hypertension, but 35.5% of them lack of tests on blood chemistry (potassium, sodium, creatinine) and urine protein. (3) 71.0% of community level hospitals could not judge the patient's "target organ damage" independently. (4) Both district and community level hospitals had all major types of anti-hypertension drugs in there pharmacy except ARB. CONCLUSION: When necessary laboratory tests for both district and community level hospitals were provided, they could accomplish the tasks of hypertension treatment and management. However, the community level hospitals should cooperate with hospitals at higher level to have a comprehensive clinical understanding of patients with high blood pressure. PMID- 14761617 TI - [An analysis on the cost of hypertensive outpatient in the community hospital in Shanghai]. AB - OBJECTIVE: To study the cost of the hypertensive outpatients. METHODS: The study randomly selected 460 insured patients with hypertension and investigated their cost on each case in the out-patient department through 2002, based on the electronic system of medical insurance. RESULTS: As a whole, the distribution of hypertensive outpatient expenditure takes on the positively skewed, with the median of 1 567.9 Yuan RMB. With the increase of age, the average expenses in each age group increased accordingly. In the study, the average number of outpatient attendances per patient was 19.5, the average expenses per visit was 115.4 Yuan RMB. In age groups 40 - 49 and 50 - 59, expenses of outpatient in male and female groups are obviously different in 2002 (Wilcoxon W(40 - 49) = 36, P(40 - 49) = 0.037; Wilcoxon W(50 - 59) = 374, P(50 - 59) = 0.023), as well as the number of out-patients (Wilcoxon W(40 - 49) = 52.5, P(40 - 49) = 0.007; Wilcoxon W(50 - 59) = 379, P(50 - 59) = 0.028). When considering the factors of gender and age at one time, the outpatient expenditures in the male group were significantly different between the different age groups (chi(2) = 22.3, P < 0.001), as well as the number of outpatients (chi(2) = 25.4, P < 0.001). In addition, the expenditure of drugs, which took a large proportion of the total expenditure of hypertensive outpatients (about 83.6 percent), was divided into three parts according to the degree of correlation with hypertension: direct expenses related to the with disease, the indirect expenses and the irrespective. The proportions of each part were 19.9 percent, 32.3 percent and 47.8 percent respectively. CONCLUSION: When economic evaluation of community prevention is carried out, the cost and cost-benefit analysis based on the analysis of outpatient expenditure and the proportion of expenses on hypertension should be taken into account. Additionally, to provide appropriate mode of medicare, to impact the behaviors and expenditure of patients, and to provide low-cost but good effective drug are also essential and important factors. PMID- 14761618 TI - [Study on the influence of knowledge about hypertension and clinical management competence among physicians treating hypertensives in district and community hospitals]. AB - OBJECTIVE: To analyze the influence of knowledge about hypertension and clinical competence among physicians in district and community hospitals on management of hypertensives. METHODS: Questionnaire investigation was used in 9 district and community hospitals in Chaoyang and Haidian district, including 181 physicians and 204 patients with hypertension. RESULTS: (1) The hospitals involved were divided into two groups according to our evaluation on the knowledge of hypertension and clinical competence of physicians. Four hospitals were graded as high-score group and 5 hospitals as low-score group. (2) There was no significant difference on physicians' evaluation between district and community hospitals. There was higher proportion of hypertensives with instructed physical exercises, reducing salt ingestion, psychological balance and weight reduction in district hospitals than those in community ones. (3) The proportion of hypertensives who were examined with funduscopy, ambulatory pressure and instructed with physical exercises, reducing salt ingestion and weight reduction in high-score group was obviously higher than that in low-score group. The control rates of blood pressure, on the days of examination during lastest check-up or the past three months, were significantly higher in high-score group than in low-score group (P < 0.05). CONCLUSION: Knowledge of hypertension and clinical management competence among physicians in district and community hospitals did influence the management of hypertension and education of physicians and thus should be increased. PMID- 14761619 TI - [Relationship between the patients' knowledge on hypertension prevention and control and the rate on blood pressure control]. AB - OBJECTIVE: To study the relationship between blood pressure control status and patients' knowledge on hypertension prevention and control among hypertensive patients. METHODS: A total of 726 hypertensives were selected from four community health service centers (2 urban and 2 rural) in Beijing. All subjects were investigated by questionnaires and their blood pressures were measured at the same time. RESULTS: The rate for blood pressure under control (< 140/90 mm Hg, 1 mm Hg = 0.133 kPa) in the rural and urban patients were 46.4% and 23.9% respectively. The control rate increased with the increase of patients' knowledge on prevention and control of hypertension in both urban and rural patients. The cumulative effect of knowledge on hypertension control status could contribute 30.0% to the difference in hypertension control rate between rural and urban patients. CONCLUSION: Patients' knowledge on hypertension control was significantly related to the rate on hypertension control. Health education should be helpful to improve the rate on hypertension control. PMID- 14761620 TI - [A cross-sectional study on knowledge and the ability of hypertension treatment among physicians in district and community hospitals]. AB - OBJECTIVE: To estimate the knowledge level and clinical ability of hypertension treatment among physicians in district and community hospitals in big and moderate cities. METHODS: 43 district and community hospitals were selected by non-random sampling in Beijing and Baotou inner-Mongolia municipality. A total of 754 physicians in those hospitals were investigated, through an examination. RESULTS: The correct rates on blood pressure stages, risk stratification and treatment strategy were 64%, 28% and 54%, respectively. The knowledge level on non-pharmacologic treatments was insufficient among physicians. The knowledge level of special indications on the common antihypertensive drugs (such as diuretics, beta-blockers, ACE inhibitors) was even worse. CONCLUSION: Physicians in district and community hospitals did not have enough knowledge and ability to fulfill the task of hypertension treatment and management. PMID- 14761621 TI - [An epidemiological study on essential hypertension in northern and western areas of China]. AB - OBJECTIVE: To understand the epidemiological characteristics of essential hypertension in the northern and western areas of China. METHODS: A community based sampling survey. RESULTS: Rates of awareness, treatment and control were 78.6%, 59.7% and 5.9%, respectively. About half of the diagnosed and treated patients took antihypertensive medicine irregularly before the survey was carried out. CONCLUSION: It is necessary to carry out education, prevention and control on hypertension and to establish a series of standards for the management and treatment on cases of hypertension. PMID- 14761622 TI - [Efficiency of the quarantine system during the epidemic of severe acute respiratory syndrome in Beijing, 2003]. AB - OBJECTIVE: An epidemic of severe acute respiratory syndrome (SARS) hit Beijing, China, between March and July 2003 with an attack rate of 1.9 per 10 thousand. (2,521 cases). To control the epidemic of SARS, a total of 30,173 residents were quarantined either in their residence or in quarantine sites. In order to understand the personal need of being quarantined and to estimate the risk of developing SARS during the quarantine period, a survey on the quarantined residents of Haidian District, Beijing, China was carried out. METHODS: 33 precincts in Haidian District divided into five groups (7 in north, 6 in south, 7 in west, 6 in east and 7 in central of Haidian District) according to the location of the precincts were involved. The director of Center of Disease Control and Prevention of Haidian District was asked to select 1 precinct from each group according to the workload of the precinct quarantine officers. From those 5 precincts we obtained lists of all quarantined persons from the precinct quarantine officers. All quarantinees were asked to complete a self-administered questionnaire. The SARS patients were diagnosed and verified according to the diagnosis criteria released by Chinese Ministry of Health which was equivalent to the SARS 'probable case' definition of WHO. All SARS cases had been reported to the relative authorities. RESULTS: By May 23, 2003, 5,186 persons had been quarantined in Haidian district, accounting for 0.23% of all residents. 1,028 of sampled quarantined residents completed the questionnaire. Of those who completed the questionnaire, 2.3% (95% CI: 1.5% - 3.5%) developed SARS while under quarantine. The median quarantine period was 14 days (range 1 day to 28 days). 61% of the quarantinees had a direct contact history with SARS patients, and all secondary SARS patients developed through contact to these quarantinees. The remaining 39% quarantinees who did not have a direct contact history with SARS patients had not developed SARS during the period under quarantine. 37% of the quarantees had direct contact during original patients' symptomatic period with an AR of 6.3%. Persons who looked after the illed SARS patient(s) during their symptomatic period, had an highest attack rate of 31% (95% CI: 20% - 44%). 63% (95% CI: 60% - 66%) of the total quarantined persons did not have direct contact with a SARS patient during his/her symptomatic period, with an attack rate of 0% (95% CI: 0% - 0.73%). CONCLUSION: Only those persons having direct contact with ill SARS patients need to be under quarantine. The overall cost for quarantine on SARS prevention could be reduced by as much as 63% if the quarantine program was limited to this group. No evidence was found that SARS patient was infective during the incubation period. PMID- 14761623 TI - [Epidemiological features of severe acute respiratory syndrome in Beijing]. AB - OBJECTIVE: To describe the epidemiologic features of severe acute respiratory syndrome (SARS) in Beijing. METHOD: Database of the 2 521 probable cases of SARS in Beijing Center for Disease Prevention and Control was used. RESULTS: The course of SARS epidemic in Beijing could be divided into five phases: import and spreading-from 1 to 31 in March, rising-from April 1 to April 15, peak-from April 16 to May 4, declining-from 5 to 18 in May, terminating-from 19 to 28 in May. The proportions of portable cases of SARS in each phase were 2.7%, 13.6%, 71.0%, 11.6% and 1.1%, respectively. Totally, 2 521 portable cases were diagnosed and verified according to the diagnostic criteria of SARS issued by the Ministry of Health. Among them, 192 died from SARS. The incidence and mortality rates of SARS were 18.57 per 100,000 and 1.41 per 100,000 with the fatality of 7.6%. The ratio of male to female with SARS was 1:0.97. The highest incidence rate of SARS was in the group of 20 - 29 years (30.85 per 100,000), and the lowest was in the group of 0 - 14 years (2.54 per 100,000). People aged 20 - 49 accounted for 72.3% of all SARS cases. The incidence rates in urban, suburb and far-suburb were 32.25/100,000, 20.57/100,000 and 8.90/100,000, respectively, decreasing according to the population density. Health care providers (17.3%), staff (12.9%), retirees (11.4%), workers (9.7%) and house-hold unemployees (8.8%) appeared to be at the five top risk populations being infected. The fatality increased significantly with age. CONCLUSION: Beijing was the most severe epidemic region of SARS in the world, but the fatality was the lowest. PMID- 14761624 TI - [Epidemiological study of rotavirus diarrhea in Beijing, China - a hospital-based surveillance from 1998 - 2001]. AB - OBJECTIVE: To provide information on epidemiology of rotavirus infection in Beijing, China. METHODS: An ongoing hospital-based surveillance was conducted among children < 5yr old with acute diarrhea according to WHO generic protocol (CID-98). During a 3-year study (Apr. 1998 to Mar. 2001), a total of 484 stool samples were collected from 1 457 patients, including 275 samples from 1 048 outpatients and 209 samples from 409 inpatients. RESULTS: The overall detection rate of rotavirus infection was 25.4%. Rotavirus was responsible for 27.3% of diarrhea inpatients on a yearly base, and 46.2% during rotavirus season. Two peaks of diarrhea were observed each year, one in the summer (June-Sep.) due to bacterial dysentery (16.7%) and another in fall winter (Oct.-Dec.) due to rotavirus infection (23.0%). The detection rate on rotavirus was the highest in age group of 6 - 11 months (38.2%), followed by 1 - 2 years old (28.5%). Ninety six point eight percentage of children were infected under 3 years of age. The number of deaths, possibly caused by rotavirus diarrhea were accounted for 40% of all diarrhea deaths and 11.1% of the total deaths. Serotyping of 123 rotavirus isolates showed that serotype G1 (55.3%) was predominant, followed by G2 (26.8%), G3 (9.8%), G4 (0.8%), and 10 isolates (8.1%) remained non-typeable. Mixed infections (0.8%) seemed to be rare. CONCLUSION: Rotavirus diarrhea was an important infectious disease among children in Beijing. Safe and effective rotavirus vaccines for the prevention of severe diarrheas and the reduction of treatment costs are of significant importance to China. PMID- 14761625 TI - [The prevalence of mild cognitive impairment among residents aged 55 or over in Chengdu area]. AB - OBJECTIVE: To study the prevalence of mild cognitive impairment (MCI) in the urban and the rural areas in Chengdu, Southwest China. METHODS: Residents aged 55 or over were selected by stratified random cluster sampling from 19 districts, cities, and counties of Chengdu area in Sichuan province. A two-stage survey was carried out. In the first stage, CMMSE, CES-D were used as screening instruments. In the second stage, Diagnostic questionnaires of dementia and CDR were used as diagnostic instruments. The diagnostic criteria of mild cognitive impairment adopted from Petersen's were: (1) memory complaint; (2) normal activities of daily living; (3) normal general cognitive function; (4) memory impairment incompatible with age; (5) not demented; (6) CDR = 0.5 and (7) exclusion of the reversible cognitive impairment caused by other factors (i.e. depression). RESULTS: Three thousand, nine hundred and ten subjects were examined. The prevalence rates of MCI was 2.4%. The MCI prevalence rates in the urban and the rural areas were 1.5%, 2.5% respectively, without significant difference. The MCI prevalence in males and females were 1.8%, 2.9% respectively. Prevalence rate in female was higher than in males with significant difference. Prevalence of illiteracy (4.0%) was the highest among different educational levels. The accumulated prevalence increased with age. CONCLUSION: The prevalence of MCI (2.4%) was slightly higher than the prevalence of AD (2.05%) in the same areas of Chengdu. MCI seemed to be a high risk factor for AD which should to be followed up. Early intervention in MCI might be helpful in the prevention of AD. PMID- 14761626 TI - [Study on the hemoglobin levels of children under the age of three years and the prevalence of anemia at high altitude in Tibet of China]. AB - OBJECTIVE: To study the hemoglobin status of children under the age of three years and the prevalence of anemia at high altitude. METHODS: Cross-sectional study and randomly sampling were used. Blood was collected on tip of finger and the level of hemoglobin was measured using B-Hemoglobin photometer. We adjusted the hemoglobin based on altitude, using three methods of CDC, Direen's and Dallman's and then estimated the prevalence of anemia. RESULTS: (1) 1 127 children were examined. The findings indicated that the average hemoglobin was about 120 g/L. There was no significant difference for Hb between boys and girls. Urban children had a higher hemoglobin level (123.3 g/L) than rural children (119.9 g/L). Analysis of covariance indicated that, after controlling the factor of altitude, the mean content of hemoglobin of rural children was lower than that of urban children and hemoglobin varied among districts. (2) Different method used for correction of hemoglobin caused different prevalence rates of anemia but they were higher than un-corrected prevalences: 90.4% for CDC method, 72.3% for Dirren's method and 65.0% for Dallman's method respectively. CONCLUSION: Higher altitude seemed to affect the levels of hemoglobin significantly. The prevalence of anemia in children might be higher, but current hemoglobin correction methods might not be suitable for correcting hemoglobin of Tibetan children. So we suggested that it was imperative to establish a relationship between altitude and hemoglobin of Tibetan children. PMID- 14761627 TI - [Study on the epidemiology and risk factors of renal calculi in special economic zone of Shenzhen city]. AB - OBJECTIVE: To study the epidemiological characters and risk factors of renal calculi in Shenzhen for future intervention. METHODS: Risk factors of renal calculi were analyzed by factor analysis and linear structural relation model, followed by fitting and evaluating the model. RESULTS: The prevalence rates of renal calculi were 8.00% and 5.12% in males and females. The results of LISEREL showed that the total effect of age and sex affecting the development of renal calculi was 0.4614, with directly effect 0.3600 and indirect 0.1014. The direct effects of blood uric acid, blood calcium and body mass index, blood cholesterol, blood sugar were 0.3400 and 0.3200 respectively. The indirect effect of education, exercise and dietary habit affected the development of renal calculi through index of biochemistry and obesity, were -0.0416 and 0.1882. CONCLUSION: Sex, age, obesity and high blood cholesterol, high blood sugar, high blood uric acid, high blood calcium were the direct influencing factors to renal calculi. At the same time, education, exercise and dietary habit were also associated with the disease. PMID- 14761628 TI - [Study on the relationship between smoking, alcohol intake and hyperlipidemia in fishermen]. AB - OBJECTIVE: To identify the relationship between smoking, alcohol intake and hyperlipidemia in fishermen. METHODS: 115 fishermen were randomly recruited and divided into case and control groups according to the result of blood lipoprotein. A questionnaire was used to record general information and the history of smoking and alcohol intake. Statistics were gathered to compare the difference of lipoprotein and apolipoprotein level between exposure and control groups and to calculate the OR value of smoking and alcohol intake. RESULTS: The OR of smoking was 3.417 (95% CI: 1.132 - 10.308), with significant dosage-effect relationship between smoking index and hyperlipidemia. The serum low density lipoprotein-cholesterol (LDL-C) and apolipoprotein (apo) B levels in smoking group was higher than that of control group. The OR value of alcohol intake at early age (early than 20) were 3.275 (95% CI: 1.249 - 8.580) and 4.016 (95% CI: 1.475 - 10.952) respectively. The LDL-C, apoB, the serum total cholesterol (TC)/high density lipoprotein-cholesterol (HDL-C) levels in alcohol abuse group were higher than that of control group. CONCLUSION: Smoking and alcohol abuse were important risk factors of hyperlipidemia, through changing the level of LDL C and apoB. There was synergistic action between smoking and alcohol abuse in the development of hyperlipidemia. PMID- 14761629 TI - [Epidemiological study of human caliciviruses among children with acute diarrhea in Lulong county, 1999 - 2001]. AB - OBJECTIVE: To investigate the epidemiological characteristus of human caliciviruses (HuCVs) among children under 5 years of age with acute diarrhea and to estimate the disease burden in Lulong county. METHODS: HuCVs were detected by enzyme linked immunosorbent assay (ELISA) and reverse transcription-polymerase chain reaction (RT-PCR). Some PCR amplicons were cloned and sequenced. Phylogenetic tree was constructed for strain characterization. The rate of HuCVs attributed hospitalization was estimated according to the positive rate of HuCVs detection in fecal specimens collected from hospitalized diarrhea patients. RESULTS: Between July 1999 and June 2001, 708 fecal specimens were collected, of which 393 rotavirus-negative and 5 rotavirus-positive specimens were detected for HuCVs. Thirty-one point six percentage of fecal specimens from patients with diarrhea was HuCVs positive. Among inpatients, HuCVs positive rate was 17.5%. HuCVs detection was mainly distributed in 3 - 17 mouth-old children, in winter. All 11 strains belonged to NLV GII in which 6 strains GII-3, 2 strains GII-4 and 3 strains GII-7, and they shared 55.1% - 100% nucleotide identity. NLV GII-4 and GII-7 were identified in 2000, while NLV GII-3 and GII-7 in 2001. The preliminary estimate of HuCVs-attributed hospitalization rate was 3.6 per thousand. CONCLUSION: Human caliciviruses with different genotypes circulated among children in Lulong county with GII NLVs were the prevalent strains. The disease burden of HuCVs was second to rotavirus. PMID- 14761630 TI - [Distribution of virulence associated genes among strains of Leptospira]. AB - OBJECTIVE: To analyze factors related to the virulence associated genes of Leptospires. METHODS: Twelve putative virulence associated genes were detected by polymerase chain reaction (PCR) method in 38 reference strains, 81 field strains of Leptospira interrogans isolated from patients or animals, and 12 avirulent strains of Leptospira biflexa. RESULTS: These putative virulent genes were widely distributed among the strains of Leptospira interrogans, but only few of them were detected in Leptospira biflexa. Gene lipL32 was detected in all strains of Leptospira interrogans. Distribution of gene lipL36 was varied significantly with detected rates from 0 to 90.91%. Gene la1608 had a positive rate of 87.50% for strains of serogroup Icterohaemorrhagiae, but was only detected in few strains of other serogroups with a range from 0 to 25.00%. Rate of detection on gene sphA was 17.65% in Leptospira interrogans, and was absent in serovar hardjo reference strain. CONCLUSION: Results indicated that these genes might be of importance for the virulence and pathogenicity of Leptospira interrogans, while gene lipL32 might be one of the common antigens. Gene lipL36 might be involved in serogroup specificity with genetic diversity, but gene la1608 was as one of the genes with specificity for serogroup Icterohaemorrhagiae. However, serovar hadjo might hold quite different genetic characteristics when compared with the other serovars of Leptospires. PMID- 14761631 TI - [A new type of spotted fever group Rickttsiaes detected in the area of Changbai mountain, Jilin province]. AB - OBJECTIVE: In order to find out the current situation of tick-borne spotted fever in the area of Changbai mountain, Jilin province. METHODS: In this study, a polymerase chain reaction (PCR) method was developed with primers R. rOmpA 190.70p and R. rOmpA 190.701n designed on the basis of rOmpA gene, which is specific for examining spotted fever group Rickttsiaes (SFGR). Six hundred nighty three ticks were tested and a positive PCR product amplified from D. silvarum specimen (named JL-02) was cloned and sequenced. RESULTS: The SFGR DNA was detected from D. silvarum, Haemaphysalis concinna with the positive rates were 53.81% and 7.41% respectively. Its nucleotide sequence of 587 bp rOmpA and derived amino-acids showed 100.00% similarity with nucleotide sequence of DnS 14 and 99.00% with DnS 28 from the Former Soviet Union according to the result of BLUST and CLUSTAL, which was differential from the DNA sequences of strains previously detected in China. CONCLUSION: The natural focus of tick-borne spotted fever did exist in the area of Changbai mountain. The DNA sequence of SFGR was similar to that of DnS 14, which was first reported in China. PMID- 14761632 TI - [Study on ribotyping of Lyme borreliosis spirochete in Guizhou province]. AB - OBJECTIVE: To define the main genotypes in Guizhou agricultural areas by molecular epidemiologic investigation of 21 Borrelia burgdorferi sensu lato of Lyme disease spirochetes and to provide the scientific bases for formulating a preventive policy. METHODS: Polymerase chain reaction (PCR) technique was used to amplify the 23S(rrl)-5S(rrf) intergenic spacer, and amplified products were analyzed by restriction fragment length polymorphism (RFLP) and nucleotide sequencing. RESULTS: There were two genospecies in the strains: 20 strains belong to Borrelia valaisiana, 1 strain is Borelia sp. CONCLUSION: Borrelia valaisiana was the main genotype in Guizhou agricultural areas. The harmness of B. valaisiana to human being has been confirmed. In order to efficiently prevent the harmness of agent to the people in Guizhou agriculture areas, we should study the risk further. PMID- 14761633 TI - [Quality of life of patients with arthritis in China]. AB - OBJECTIVE: To evaluate the quality of life of arthritis patients in China. METHODS: Patients (n = 1 344) were selected by stratified random sampling according to the economic conditions and the ranking of hospitals, and asked to complete the arthritic self-reported questionnaire of quality of life. RESULTS: The scores of quality of life were not different between male arthritis and female arthritis in different age groups (P > 0.05). The lower scores of quality of life were reported more often among the women older than 65 years. Those of lower educational level and lower income got fewer scores. Compared to the married and unmarried patients, the divorcees and those who were bereft of their spouses gained low scores (t = 9.310, P = 0.001). Among different occupation groups, the labors and farmers also got lower scores than others (P < 0.05). On the other hand, if the arthritis with better physical conditions, it was more possible that the scores were higher, but to pressed pain index, there were no different scores shown between the second and third grade (mean difference = 4.910, P = 0.765), and to joint swell index, there are no different scores between either the 0 and 1 grade (mean difference = 16.308, P = 0.079) or the 2 and 3 grade (mean difference = 6.643, P = 0.533). CONCLUSION: To improve the quality of life of all arthritis patients, enhancing the quality of medical technology and enriching the medical knowledge are evidently necessary. However, it is also important to prefect social security system, to improve the country education and pay attention to life of elderly in order to improve the quality of life of all arthritis patients. PMID- 14761634 TI - [Study on the prevalence of arthritis and relevant factors in Shanghai]. AB - OBJECTIVE: This study aimed to understand the prevalence rate, epidemiological characteristics and relevant factors of arthritis in Shanghai. METHODS: A sample of 7 575 residents aged 15 years and above was drawn from 6 communities under multiple stage cluster sampling. A household survey with questionnaire was carried out to differentiate both undiagnosed patients and those with definite arthritis. Those who had not been diagnosed before were asked to carry further clinical examinations by a rheumatologist. RESULTS: The prevalence rate of arthritis was 6.11%, including osteoarthritis (OA) 4.18%, rheumatoid arthritis (RA) 0.52%, gout 0.28%, ankylosing spondylitis (AS) 0.28%, rheumatic arthritis 0.49% and other types arthritis 0.82%. Arthritis was significantly related to cardiovascular disease, pulmonary disease and gastrointestinal disease. Age, female and obesity might serve as risk factors for arthritis. Physical labors and living in rural area might have protecting effects. CONCLUSION: Elderly and female seemed to be at high risk for arthritis. Weight control and more exercise should be encouraged to reduce the risks. For arthritis patients, treatment to other chronic diseases should not be ignored. PMID- 14761635 TI - [Quantitative assessment of risks on cerebral vascular diseases in urban residents in Sichuan]. AB - OBJECTIVE: To set a quantitative criteria for determining the risks on cerebral vascular disease (CVD) so to identify that potential risk of an individual dying from CVD and to predict the individual risk of CVD. METHODS: Data on case-control and cohort studies published during 1978 to 2003 was collected through retrieval of literatures, and data on surveillance of behavior exposure was provided by Chengdu Municipal Center for Disease Control and Prevention. Pooled odds ratio (OR) and relative risk (RR) of all risk factors for CVD were estimated using software for meta-analysis to enable the varied levels of risk factors be converted into risk fractions by statistical models. RESULTS: A risk score conversion table (quantitative criteria for assessment) of main risk factors for CVD was developed for men and women aged 35 - 69 at an interval of five years, including smoking, passive smoking, hypertension, high blood cholesterol levels, body mass index, lack of physical activity, alcohol drinking, dietary fat consumption, milk intake, oral contraceptive use, past history of diabetes and CVD, family history of CVD etc. Individuals with all these risk factors had a risk score beyond 1.00, but was equal to or below 1.00 when without. The risk score would increase along with the rise of one's risk level. CONCLUSION: Estimation of risk of dying from CVD was based on risk score conversion table of risk factors for CVD, which could be used to predict individual potential risk of dying from CVD in the following 10 years. Our data provides evidence that education to be strengthened to persuade people to change their unhealthy lifestyles and behaviors. PMID- 14761636 TI - [Reliability of measurement and the methods of estimating reliability]. AB - OBJECTIVE: To explore the methods of estimating measurement reliability and their limitations. METHODS: According to the given examples, reliabilities of measurement were calculated and limitations of the methods of estimating reliability were analyzed. RESULTS: The Kappa value of interobserver reliability was 0.793 between two pathologists. Between the two populations with different prevalence rates, the values of Kappa were 0.800 and 0.137 respectively, and with big difference. Cronbach's alpha coefficient of compositive index for A type behavior was 0.55. CONCLUSION: The Kappa index and alpha coefficient were both inherently population-specific. Before generalizing to different populations, the reliability needs to be measured. PMID- 14761637 TI - [Clinical observation of the effect of tangential excision within 24 postburn hours on the patients with deep partial thickness burn]. AB - OBJECTIVE: To investigate the safety and the clinical effect of the tangential excision of the patients with deep partial thickness burn within 24 postburn hours (PBHs). METHODS: Twelve patients with deep partial thickness burn with the indication for tangential excision, the operation was carried out within 24 PBHs. These patients were designated as group A. Another group of fourteen patients with similar conditions undergoing tangential excision during 4 - 6 PBDs were designated as group B. The amount of fluid infusion during shock stage, the shock signs, the vital signs during recovery period, the urine output, as well as the healing time were compared between the two groups. RESULTS: There was no obvious difference in the amount of fluid infusion and the shock signs between the two groups. There were evident differences in the body temperature and heart rate during recovery period, with the urine output increased dramatically during shock stage in group A when compared with those in group B (P < 0.05-0.01). The average wound healing days in group A were less than that in group B (P < 0.01). CONCLUSION: Tangential excision within 24 PBHs was applicable and safe for the patients with deep partial thickness burn, and wound healing time was thus shortened. PMID- 14761638 TI - [An experimental study on the repair of the necrotic tubular bone with pedicled vascular periosteum grafting in rabbits]. AB - OBJECTIVE: To investigate the mechanism of the repair of the necrotic tubular bone with pedicled vascular periosteum grafting in rabbits. METHODS: Twenty mature New Zealand rabbits aged 8 months were randomly divided into test and control groups by placebo-controlled protocol. A segment of radius measuring 1.2 cm in length with pedicled vascular periosteum of the rabbits in the test group was obtained from the middle distal part of the bone, and boiled for 30 minutes. The cooled bone was then re-grafted and wrapped with vascularized periosteum and immobilized, while those in the control group, no periosteal grafting was done. X ray examinations of the bones were carried out at 2, 4, 6, 8 approximately 10 post operation weeks (POWs). Meanwhile 4-5 rabbits were sacrificed for the harvesting of the radius on both sides for histological examination. RESULTS: It was revealed by X-ray and histological study that the periosteal hyperplasia could be observed 2-4 weeks after operation in the test group. The sign of union of the fracture and the formation and reconstitution of the trabeculae could be observed 6-8 weeks after operation. The reconstitution of reticulated bone, the formation of the lamellar bone and new haversian system could be observed during 8-10 POW. The structure of the new formed bones became more mature and the fracture healed at 12 POW. In contrast, there was no changes in the necrotic bones in the control group at 2 POW and no signs of fracture healing at 4-6 POW, and the defect of the bones could be observed at 8 POW. The necrotic bones were totally dissolved and absorbed, and the bone defects could be observed clearly filled with fibrotic tissues at 12 POW. CONCLUSION: The necrotic bones could be replaced by regeneration of new bone originated from the pedicled vascular periosteum, so that the necrotic tubular bones could be repaired. But the necrotic bones in the control group were eventually absorbed, resulting in bone defects. PMID- 14761639 TI - [In vitro amplification and identification of immature dendritic cells from murine bone marrow]. AB - OBJECTIVE: To establish a method for in vitro amplification of immature dendritic cells from murine bone marrow, and to identify it with morphological, immunological phenotype determination, and functional examination. METHODS: Dendritic cells from murine bone marrow were cultured with different dosage of rmGM-CSF. The suspending cells were examined with scanning electronic microscope, and the non-sensitized T lymphocyte proliferation was observed by mixed lymphocyte reaction. RESULTS: Dendritic cells (DC) cultured in lower dosage of rmGM-CSF (Gm(low)DC) exhibited typical characteristics of DCs with high expression of CD11c and low expression of CD40 and I-A/I-E, and non-expression of B7-1 on the surface of the cells. The capacity of Gm(low)DC to stimulate the proliferation of non-sensitized T lymphocyte in vitro was weaker than that of Gm(high)DC. CONCLUSION: Gm(low)DC exhibited typical characteristics of DC, immature in cell phenotype and cell functions, suggesting that our methods of immature DCs culturing was feasible. The dosage of rm GM-CSF has direct relationship with the maturation degree of DC. Generally speaking, mature DC was mainly induced by high dosage of rmGM-CSF, while immature DC by low dosage. PMID- 14761640 TI - [Effects of basic fibroblast growth factor on the matrix metalloproteinase-2,7 and its tissue inhibitor on deep partial thickness burn wounds in rats]. AB - OBJECTIVE: To observe the changes in matrix metalloproteinase-2,7 (MMP-2,7) and tissue inhibitor of metalloproteinase-2 (TIMP-2) in deep partial thickness burn during the process of wound healing, and the effects of bFGF on wound healing. METHODS: The rats inflicted by 30% TBSA deep partial thickness burn were randomly divided into simple scald and bFGF treatment groups. Biopsies from wound skin were harvested at 3 and 6PBHs and 1, 3, 7, 14 PBDs for the detection of the epithelialization rate and collagen content. The above indices were also detected in the skin of another 6 normal rats as normal control. RESULTS: (1) The epithelialization rate in bFGF treatment group was higher than that in simple scald group during 3PBH to 14 PBD. (2) The collagen contents in both bFGF treatment group and simple scald group were continually decreased during 3 PBH to 3 PBD, and increased from 7 to 14 PBD, but still lower than that in normal control (P < 0.05). (3) The expression of MMP-2,7 and TIMP-2 in simple scald group enhanced from 1 to 14 PBD, and peaked on 7 PBD. (4) The expression of MMP 2,7 in bFGF treatment group was similar to that in simple scald group from 3 to 6 PBH, while the expressions of MMP-2,7 and TIMP-2 was higher than those in simple scald group from 1 to 14 PBD. CONCLUSION: The collagen deposition would be affected by the activities of extracellular matrix in scald wound in rats. Changes in MMP-2,7 and TIMP-2 expressions were an important process of wound repair, which was closely related to the acceleration of wound healing by the application of bFGF. PMID- 14761641 TI - [Comparison of the effects of rhEGF with rhbFGF on the acceleration of wound healing]. AB - OBJECTIVE: To investigate the mechanism and the accelerating effect of rhEGF and rhbFGF on wound healing. METHODS: Twelve New Zealand rabbits with 72 incised wounds on ventral side of 24 ears were randomly divided into two therapeutic groups (rhEGF of 10 ug/cm(2) and rhbFGF of 100 AU/cm(2)) and a control group (1% silver sulfadiazine cream, SD-Ag). The general conditions of the wound healing was observed grossly. Biopsies were harvested at different time points for the pathomorphological examination, the electron microscopic examination, and for assessment of integrin beta1 mRNA expression by in situ hybridization. RESULTS: The expressions of integrin beta 1 mRNA in two therapeutic groups were significantly higher than that of control group. The quality of the wound healing was improved in therapeutic group with its healing time shortened when compared with that in control group (P < 0.05). There was an obvious difference in the number of fibroblasts and capillary gemmules between the therapeutic and control groups (P < 0.05). CONCLUSION: The wound healing and quality could be improved by both rhEGF and rhbFGF, but rhbFGF seemed better to be employed during the early and middle stages of the wound repair for the growth of granulation tissue, while rhEGF should be applied at the late stage of wound repair to accelerate the re epithelialization of the wound. Combined application of rhEGF with rhbFGF according to time effect could be more beneficial to the wound repair. PMID- 14761642 TI - [A preliminary study on the identification and distribution of epidermal stem cells in different degrees of burn wounds in scalded rats]. AB - OBJECTIVE: To investigate the distribution of epidermal stem cells (ESCs) in different degrees of burn wounds in scalded rats. METHODS: Thirty-two Sprague Dawley (SD) rats were employed in the study. First degree (I), shallow (shallow II) and deep partial thickness (deep II) and full thickness burn wounds (III) were created on the rat skin. Burn wound samples were harvested at 24 postburn hours (PBHs) from all the wounds and were processed to tissue slices. The tissue slices were stained by immunohistochemistry technique. The expression and distribution of ESCs in different degrees of burn wounds were observed with integrins alpha 2 beta 1 and keratin 10 (K10) as first antibodies. RESULTS: K10 positive cells were found to distribute in the strata spinosum, granulosum and lucidum in the first degree burn wound (I) with large amounts of integrins alpha 2 beta 1 positive cells in the residual basal layer and skin appendages (hair follicles) in shallow partial thickness burn wound (shallow II degree), and there were less integrins alpha 2 beta 1 positive cells in the remaining skin appendages in deep dermis in deep partial thickness burn wound (deep II degree). Finally, integrins alpha 2 beta 1 positive cells were sparsely found in the III degree burn wound. CONCLUSION: The distribution of ESCs in burn wounds was closely related to the depth of burn wound. The residual ESCs might be the origin of burn wound regeneration and reepithelization. PMID- 14761643 TI - [Postburn change in the plasma level of neuron specific enolase in burned patients with cerebral malfunction]. AB - OBJECTIVE: To explore the relationship between the change in neuron specific enolase (NSE) and brain malfunction in burned patients. METHODS: The serum samples of 11 burned patients with brain dysfunction were collected for the development of the serum level of neuron specific enolase with radioimmunoassay, and the correlation between condition of systemic inflammation and the levels of neuron specific enolase was assessed. RESULTS: The level of NSE in burn patients with cerebral malfunction was obviously higher than that in control, and the level was correlated with the systemic inflammation. CONCLUSION: The change in the level of serum NSE could reflect the damage degree of central nervous system to some extent. PMID- 14761644 TI - [Early repair of full layer eyelid defect caused by chemical burn]. AB - OBJECTIVE: To investigate the optimal time and method of the early repair of the full layer eyelid defect caused by chemical burn. METHODS: Free nasal septum mucosal cartilage flap with muscle flap, skin grafting, or skin flap were performed in 18 cases (19 eyelids) with chemical burn within 4 postburn weeks. Eyelid reconstruction and corneal transplantation were performed at the same time in 4 patients. RESULTS: All the reconstructed eyelids and transplanted cornea survived. The incidence of severe complications, such as exposure keratitis, corneal ulcer and eyeball perforation decreased. CONCLUSION: Full layer eyelid defect caused by chemical burn should receive early reconstruction and repair, including timely reconstruction of eyelid for the sake of protecting the eyesight and of alleviating the inflammatory reactions, and the corneal transplantation should be done at the same time to avoid corneal perforation. Nasal septum mucosal cartilage flap could be ideal for the eyelid reconstruction. PMID- 14761645 TI - [Clinical study on the immunoregulation effects of cytokines on the acellular xenogenic dermal matrix]. AB - OBJECTIVE: To investigate the relationship between some cytokines and early outcome of grafts in burn patients receiving xeno-/allo-ADM grafting. METHODS: Nineteen xeno-ADMs were grafted onto the wounds of 12 patients after escharectomy in extremities, and 18 allo-ADMs were grafted onto the escharectomy wounds of 15 patients in extremities. All the grafts were covered with thin split thickness skin autografts. Six patients grafted with split thickness skin autografts (auto TTS) were employed as control. After the grafts survived for 4 to 8 weeks, immunohistochemistry and ELISA methods were employed to determine the contents of IL-1 beta, IL-4, IL-6, TNF-alpha and IFN-gamma in the exudation fluid from wounds after the rejection of xeno-ADM, local skin and peripheral blood. RESULTS: It was exhibited by immunochemistry staining that the positive cellular density and staining intensity of the IL-1 beta, IL-4, IL-6, TNF-alpha and IFN-gamma in the grafts were ranked as following: Xeno-ADM > allo-ADM > auto-TTS (P < 0.05 - 0.01). The levels of IL-4, IL-6, TNF-alpha and IFN-gamma in the exudation fluid during the rejection of xeno-ADM were obviously higher than those in the blood of the patients as determined by ELISA, while the serum levels of IL-4 and IFN-gamma in xeno-ADM group were lower and higher than those in xeno-ADM when without rejection respectively. The serum levels of IL-4, TNF-alpha and IFN-gamma in xeno ADM group were significantly higher than those in allo-ADM and auto-TTS groups (P < 0.05 - 0.01). CONCLUSION: Local detection of high levels of IL-1 beta, IL-4, IL 6, TNF-alpha and IFN-gamma might be related to the immune augmentation mediated by cytotoxic lymphocytes and cytokines. The dynamic changes of these cytokines might be helpful to the explanation of the bad outcome of xeno-ADMs. PMID- 14761646 TI - [Study on the injurious effect of a self designed micro-skin machine on the epithelia]. AB - OBJECTIVE: To observe the injury on micro-skin induced by a self designed micro skin machine. METHODS: Micro-skin was produced either with the machine or by hand. Cells at the edge of micro-skin were observed by transmission electron microscope. succinic dehydrogenase activity in supernatant of cultivated cells was analyzed, and the cell proliferation of micro-skin was assessed by (3)H-TdR. Twenty patients were enrolled in the study for the observation of the wound healing time between the two groups of micro-skin after being grafted. RESULTS: Transmission electron microscope examination revealed that the cellular injury at the edge of the micro-skin in machine-made group was mild compared with that in man-made group. (3)H-TdR rate was elevated but the activity of succinic dehydrogenase in the supernatant of cultured cells decreased in supernatant of cultured cells of machine produced micro-skin. Wound healing time was shortened in machine made group. (P < 0.05). CONCLUSION: The cellular injury at the edge of micro-skin in the machine made group was mild when compared with that in the man made group with cell proliferation accelerated and wound healing time shortened. PMID- 14761647 TI - [Repair of murine full skin loss with composite skin of collagen scaffold containing living cells]. AB - OBJECTIVE: To construct composite skin containing living cells and to observe its significance in the repair of full skin loss in mice. METHODS: The dermal substitute was formed by culturing heterogeneous fibroblasts on the acellular chondrocyte collagen scaffold for 3 days, and then cultured on the epithelial membrane for another 10 days, to form the composite skin containing living cells. The composite skin was grafted onto full layer skin defect. The growth condition was observed and biopsies were harvested for histologic examination. RESULTS: Both fibroblasts and stratified epithelium grew well in the collagen scaffold. The composite skin adhered tightly to wounds of the mice, with obvious vascularization one week after grafting. The grafts began to merge with the wound margin at 6 post operation weeks without obvious signs of rejection. CONCLUSION: Full skin loss could be repaired by composite skin formed by acellular chondrocyte collagen scaffold as a skin substitute. PMID- 14761648 TI - [Repair of contracture deformities of the patients with extremely limited donor skin at late postburn stage]. AB - OBJECTIVE: To explore new methods to repair postburn contracture deformities in patients with extremely limited donor skin. METHODS: Five severely burned patients with extremely limited donor skin but severe deformities were enrolled in the study. The mature and the pliable scarred skin was utilized as the donor site for reconstruction of the postburn deformities. Split-thickness scarred skin was harvested for repair of postburn deformities after the scarred skin was expanded by expander, while thin razor-thin scarred skin with allo-acellular dermal matrix (ADM) was employed for the repair of postburn deformities when skin expansion was not feasible. RESULTS: All the expanded scarred skin and composite skin grafts survived completely with good function and configuration. The long term follow-up result was satisfactory, and the grafted skin was similar to that with split-thickness skin grafting. CONCLUSION: It is feasible to employ various split-thickness scarred skin for the reconstruction of postburn deformities. This technique is a new effective procedure for the reconstruction of postburn deformities, especially for those with extreme scarcity of donor site. PMID- 14761649 TI - High mobility of vesicles supports continuous exocytosis at a ribbon synapse. AB - BACKGROUND: Most synapses release neurotransmitter as transient pulses, but ribbon synapses of sensory neurons support continuous exocytosis in response to maintained stimulation. We have investigated how the movement and retrieval of vesicles might contribute to continuous exocytosis at the ribbon synapse of retinal bipolar cells. RESULTS: Using a combination of total internal reflection fluorescence microscopy and fluorescence recovery after photobleaching, we found that the great majority of vesicles within 50-120 nm of the plasma membrane move in a random fashion with an effective diffusion coefficient of approximately 1.5 x 10(-2) microm(2) s(-1). Using confocal microscopy, we found that vesicles are similarly mobile across the whole terminal and that this motion is not altered by calcium influx or the actin cytoskeleton. We calculated that the cytoplasmic reservoir of approximately 300,000 vesicles would generate about 900 vesicle collisions/s against ribbons and 28,000 collisions/s against the surface membrane. The efficient resupply of vesicles to ribbons was confirmed by electron microscopy. A 1 min depolarization, releasing 500-1000 vesicles/s, caused a 70% reduction in the number of vesicles docked at the active zone without reducing the number of vesicles attached to ribbons or remote areas of the plasma membrane. These sites were not repopulated by retrieved vesicles because 80-90% of the recycled membrane was taken up into cisternae that pinched off from the surface. CONCLUSIONS: These results indicate that the random motion of cytoplasmic vesicles provides an efficient supply to the ribbon and plasma membrane and allows the maintenance of high rates of exocytosis without an equally rapid recycling of vesicles. The selective depletion of vesicles docked under ribbons suggests that the transfer of vesicles to the active zone limits the rate of exocytosis during maintained stimulation. PMID- 14761650 TI - The anterior-posterior axis emerges respecting the morphology of the mouse embryo that changes and aligns with the uterus before gastrulation. AB - BACKGROUND: When the anterior-posterior axis of the mouse embryo becomes explicit at gastrulation, it is almost perpendicular to the long uterine axis. This led to the belief that the uterus could play a key role in positioning this future body axis. RESULTS: Here, we demonstrate that when the anterior-posterior axis first emerges it does not respect the axes of the uterus but, rather, the morphology of the embryo. Unexpectedly, the emerging anterior-posterior axis is initially aligned not with the long, but the short axis of the embryo. Then whether the embryo develops in vitro or in utero, the anterior-posterior axis becomes aligned with the long axis of embryo just prior to gastrulation. Of three mechanisms that could account for this apparent shift in anterior-posterior axis orientation-cell migration, spatial change of gene expression, or change in embryo shape-lineage tracing studies favor a shape change accompanied by restriction of the expression domain of anterior markers. This property of the embryo must be modulated by interactions with the uterus as ultimately the anterior-posterior and long axes of the embryo align with the left-right uterine axis. CONCLUSIONS: The emerging anterior-posterior axis relates to embryo morphology rather than that of the uterus. The apparent shift in its orientation to align with the long embryonic axis and with the uterus is associated with a change in embryo shape and a refinement of anterior gene expression pattern. This suggests an interdependence between anterior-posterior gene expression, the shape of the embryo, and the uterus. PMID- 14761651 TI - Initiation of gastrulation in the mouse embryo is preceded by an apparent shift in the orientation of the anterior-posterior axis. AB - BACKGROUND: It is generally assumed that the migration of anterior visceral endoderm (AVE) cells from a distal to a proximal position at embryonic day (E)5.5 breaks the radial symmetry of the mouse embryo, marks anterior, and conditions the formation of the primitive streak on the opposite side at E6.5. Transverse sections of a gastrulating mouse embryo fit within the outline of an ellipse, with the primitive streak positioned at one end of its long axis. How the establishment of anterior-posterior (AP) polarity relates to the morphology of the postimplantation embryo is, however, unclear. RESULTS: Transverse sections of prestreak E6.0 embryos also reveal an elliptical outline, but the AP axis, defined by molecular markers, tends to be perpendicular to the long axis of the ellipse. Subsequently, the relative orientations of the AP axis and of the long axis change so that when gastrulation begins, they are closer to being parallel, albeit not exactly aligned. As a result, most embryos briefly lose their bilateral symmetry when the primitive streak starts forming in the epiblast. CONCLUSIONS: The change in the orientation of the AP axis is only apparent and results from a dramatic remodeling of the whole epiblast, in which cell migrations take no part. These results reveal a level of regulation and plasticity so far unsuspected in the mouse gastrula. PMID- 14761652 TI - Awareness modifies the skill-learning benefits of sleep. AB - Behind every skilled movement lies months of practice. However, practice alone is not responsible for the acquisition of all skill; performance can improve between, not just within, practice sessions. An important principle shaping these offline improvements may be an individual's awareness of learning a new skill. New skills, such as a sequence of finger movements, can be learned unintentionally (with little awareness for the sequence, implicit learning) or intentionally (explicit learning). We measured skill in an implicit and explicit sequence-learning task before and after a 12 hr interval. This interval either did (8 p.m. to 8 a.m.) or did not (8 a.m. to 8 p.m.) include a period of sleep. Following explicit sequence learning, offline skill improvements were only observed when the 12 hr interval included sleep. This overnight improvement was correlated with the amount of NREM sleep. The same improvement could also be observed in the evening (with an interval from 8 p.m. to 8 p.m.), so it was not coupled to retesting at a particular time of day and cannot therefore be attributed to circadian factors. In contrast, in the implicit learning task, offline learning was observed regardless of whether the 12 hr interval did or did not contain a period of sleep. However, these improvements were not observed with only a 15 min interval between sessions. Therefore, the practice available within each session cannot account for these skill improvements. Instead, sufficient time is necessary for offline learning to occur. These results show a behavioral dissociation, based upon an individual's awareness for having learned a sequence of finger movements. Offline learning is sleep dependent for explicit skills but time dependent for implicit skills. PMID- 14761653 TI - Migration of the plastid genome to the nucleus in a peridinin dinoflagellate. AB - Dinoflagellate algae are important primary producers and of significant ecological and economic impact because of their ability to form "red tides". They are also models for evolutionary research because of an unparalleled ability to capture photosynthetic organelles (plastids) through endosymbiosis. The nature and extent of the plastid genome in the dominant perdinin-containing dinoflagellates remain, however, two of the most intriguing issues in plastid evolution. The plastid genome in these taxa is reduced to single-gene minicircles encoding an incomplete (until now 15) set of plastid proteins. The location of the remaining photosynthetic genes is unknown. We generated a data set of 6,480 unique expressed sequence tags (ESTs) from the toxic dinoflagellate Alexandrium tamarense (for details, see the Experimental Procedures in the Supplemental Data) to find the missing plastid genes and to understand the impact of endosymbiosis on genome evolution. Here we identify 48 of the non-minicircle-encoded photosynthetic genes in the nuclear genome of A. tamarense, accounting for the majority of the photosystem. Fifteen genes that are always found on the plastid genome of other algae and plants have been transferred to the nucleus in A. tamarense. The plastid-targeted genes have red and green algal origins. These results highlight the unique position of dinoflagellates as the champions of plastid gene transfer to the nucleus among photosynthetic eukaryotes. PMID- 14761654 TI - Kinesin II mediates Vg1 mRNA transport in Xenopus oocytes. AB - The subcellular localization of specific mRNAs is a widespread mechanism for regulating gene expression. In Xenopus oocytes microtubules are required for localization of Vg1 mRNA to the vegetal cortex during the late RNA localization pathway. The factors that mediate microtubule-based RNA transport during the late pathway have been elusive. Here we show that heterotrimeric kinesin II becomes enriched at the vegetal cortex of stage III/IV Xenopus oocytes concomitant with the localization of endogenous Vg1 mRNA. In addition, expression of a dominant negative mutant peptide fragment or injection of a function-blocking antibody, both of which impair the function of heterotrimeric kinesin II, block localization of Vg1 mRNA. We also show that exogenous Vg1 RNA or Xcat-2, another RNA that can use the late pathway, recruits endogenous kinesin II to the vegetal pole and colocalizes with it at the cortex. These data support a model in which kinesin II mediates the transport of specific RNA complexes destined for the vegetal cortex. PMID- 14761655 TI - Heparan sulfate proteoglycan syndecan promotes axonal and myotube guidance by slit/robo signaling. AB - Slit, the ligand for the Roundabout (Robo) receptors, is secreted from midline cells of the Drosophila central nervous system (CNS). It acts as a short-range repellent that controls midline crossing of axons and allows growth cones to select specific pathways along each side of the midline. In addition, Slit directs the migration of muscle precursors and ventral branches of the tracheal system, showing that it provides long-range activity beyond the limit of the developing CNS. Biochemical studies suggest that guidance activity requires cell surface heparan sulfate to promote binding of mammalian Slit/Robo homologs. Here, we report that the Drosophila homolog of Syndecan (reviewed in ), a heparan sulfate proteoglycan (HSPG), is required for proper Slit signaling. We generated syndecan (sdc) mutations and show that they affect all aspects of Slit activity and cause robo-like phenotypes. sdc interacts genetically with robo and slit, and double mutations cause a synergistic strengthening of the single-mutant phenotypes. The results suggest that Syndecan is a necessary component of Slit/Robo signaling and is required in the Slit target cells. PMID- 14761656 TI - Independent origins of Indian caste and tribal paternal lineages. AB - The origins of the nearly one billion people inhabiting the Indian subcontinent and following the customs of the Hindu caste system are controversial: are they largely derived from Indian local populations (i.e. tribal groups) or from recent immigrants to India? Archaeological and linguistic evidence support the latter hypothesis, whereas recent genetic data seem to favor the former hypothesis. Here, we analyze the most extensive dataset of Indian caste and tribal Y chromosomes to date. We find that caste and tribal groups differ significantly in their haplogroup frequency distributions; caste groups are homogeneous for Y chromosome variation and more closely related to each other and to central Asian groups than to Indian tribal or any other Eurasian groups. We conclude that paternal lineages of Indian caste groups are primarily descended from Indo European speakers who migrated from central Asia approximately 3,500 years ago. Conversely, paternal lineages of tribal groups are predominantly derived from the original Indian gene pool. We also provide evidence for bidirectional male gene flow between caste and tribal groups. In comparison, caste and tribal groups are homogeneous with respect to mitochondrial DNA variation, which may reflect the sociocultural characteristics of the Indian caste society. PMID- 14761657 TI - Diverse substrate recognition mechanisms for rhomboids; thrombomodulin is cleaved by Mammalian rhomboids. AB - The rhomboids are a recently discovered family of intramembrane proteases that are conserved across evolution. Drosophila was the first organism in which they were characterized, where at least Rhomboids 1-3 activate EGF receptor signaling by releasing the active forms of EGF-like growth factors. Subsequent work has begun to shed light on the role of these proteases in bacteria and yeast, but nothing is known about the function of rhomboids in vertebrates beyond evidence that the subclass of mitochondrial rhomboids is conserved. Here, we report that the anticoagulant cell-surface protein thrombomodulin is the first mammalian protein to be a rhomboid substrate in a cell culture assay. The thrombomodulin transmembrane domain (TMD) is cleaved only by vertebrate RHBDL2-like rhomboids. Thrombomodulin TMD cleavage is directed not by sequences within the TMD, as is the case with Spitz but by its cytoplasmic domain, which, at least in some contexts, is necessary and sufficient to determine cleavage by RHBDL2. These data suggest that thrombomodulin could be a physiological substrate for rhomboid. Moreover, the discovery of a second mode of substrate recognition by rhomboids implies mechanistic diversity in this family of intramembrane proteases. PMID- 14761658 TI - Age-specific acceleration of cancer. AB - One of the great challenges of cancer research is to explain the epidemiological patterns of cancer incidence based on the molecular processes that lead to uncontrolled cellular proliferation. The epidemiological data demonstrate that the age-specific incidence of many cancers increases in an approximately linear way with age when plotted on a log-log scale, with different slopes for different cancers. However, those epidemiological data also show that cancers of various tissues depart from log-log linearity in particular ways. Here, I illustrate those departures from log-log linearity by introducing plots of the age-specific acceleration of cancer. I then develop a very general model of cancer progression, which I use to explain the observed differences between tissues in age-specific acceleration. In one application of the model, I show that the spectacular rise and fall in age-specific acceleration observed in prostate cancer may be explained by multiple rounds of clonal expansion. In a second application, I demonstrate that the steady decline in age-specific acceleration of breast cancer may occur because precancerous mutations accumulate in many cellular lineages. PMID- 14761659 TI - Stabilized structure from motion without disparity induces disparity adaptation. AB - 3D structures can be perceived based on the patterns of 2D motion signals. With orthographic projection of a 3D stimulus onto a 2D plane, the kinetic information can give a vivid impression of depth, but the depth order is intrinsically ambiguous, resulting in bistable or even multistable interpretations. For example, an orthographic projection of dots on the surface of a rotating cylinder is perceived as a rotating cylinder with ambiguous direction of rotation. We show that the bistable rotation can be stabilized by adding information, not to the dots themselves, but to their spatial context. More interestingly, the stabilized bistable motion can generate consistent rotation aftereffects. The rotation aftereffect can only be observed when the adapting and test stimuli are presented at the same stereo depth and the same retinal location, and it is not due to attentional tracking. The observed rotation aftereffect is likely due to direction-contingent disparity adaptation, implying that stimuli with kinetic depth may have activated neurons sensitive to different disparities, even though the stimuli have zero relative disparity. Stereo depth and kinetic depth may be supported by a common neural mechanism at an early stage in the visual system. PMID- 14761660 TI - Consolidation of dynamic motor learning is not disrupted by rTMS of primary motor cortex. AB - Motor skills, once learned, are often retained over a long period of time. However, such learning first undergoes a period of consolidation after practice. During this time, the motor memory is susceptible to being disrupted by the performance of another motor-learning task. Recently, it was shown that repetitive transcranial magnetic stimulation (rTMS) over the primary motor cortex could disrupt the retention of a newly learned ballistic task in which subjects had to oppose their index finger and thumb as rapidly as possible. Here we investigate whether the motor cortex is similarly involved during the consolidation that follows learning novel dynamics. We applied rTMS to primary motor cortex shortly after subjects had either learned to compensate for a dynamic force field applied to their index finger or learned a ballistic finger abduction task. rTMS severely degraded the retention of the learning for the ballistic task but had no effect on retention of the dynamic force-field learning. This suggests that, unlike learning of simple ballistic skills, learning of dynamics may be stored in a more distributed manner, possibly outside the primary motor cortex. PMID- 14761661 TI - The ventriloquist effect results from near-optimal bimodal integration. AB - Ventriloquism is the ancient art of making one's voice appear to come from elsewhere, an art exploited by the Greek and Roman oracles, and possibly earlier. We regularly experience the effect when watching television and movies, where the voices seem to emanate from the actors' lips rather than from the actual sound source. Originally, ventriloquism was explained by performers projecting sound to their puppets by special techniques, but more recently it is assumed that ventriloquism results from vision "capturing" sound. In this study we investigate spatial localization of audio-visual stimuli. When visual localization is good, vision does indeed dominate and capture sound. However, for severely blurred visual stimuli (that are poorly localized), the reverse holds: sound captures vision. For less blurred stimuli, neither sense dominates and perception follows the mean position. Precision of bimodal localization is usually better than either the visual or the auditory unimodal presentation. All the results are well explained not by one sense capturing the other, but by a simple model of optimal combination of visual and auditory information. PMID- 14761664 TI - Regulation of cardiac myocyte apoptosis by the GATA-4 transcription factor. AB - Apoptosis of cardiac muscle cells plays important roles in the development of various heart diseases including myocardial infarction and anthracycline-induced cardiomyopathy. Understanding the regulatory mechanisms of cardiac myocyte apoptosis and survival is important for establishing therapeutic strategies against heart disease. Our recent experiments demonstrate that the GATA-4 transcription factor not only mediates cardiac hypertrophy, but also regulates apoptosis and survival of adult cardiac muscle cells. Apoptosis induced by anthracyclines is associated with decreased expression of GATA-4, while the restoration of GATA-4 levels via ectopic expression attenuated the apoptosis. Survival factors of cardiac myocytes such as hepatocyte growth factor and endothelin-1 activate GATA-4, and this signal transduction mechanism at least in part serves to protect the heart against oxidative stress. PMID- 14761665 TI - Anti-inflammatory activity of tea (Camellia sinensis) root extract. AB - Pharmacological studies were carried out with methanol-water (1:1) extract of dried tea (Camellia sinensis) root extract (TRE). TRE was found to possess anti inflammatory, analgesic and antipyretic activities at 1/10th of its LD50 dose of 100 mg/kg i.p. It was found that TRE inhibited the arachidonic acid-induced paw oedema in rats which indicated that TRE produced the anti-inflammatory activity by inhibiting both the cyclooxygenase and lypooxygenase pathways of arachidonic acid metabolism. TRE also enhanced peritoneal cell count and the number of macrophages in normal mice. It is plausible that the saponins present in TRE may be responsible for these activities of TRE. PMID- 14761666 TI - Effects of subacute treatment with benzopyranopyrimidines in hemostasis and experimental thrombosis in mice. AB - The antithrombotic activity of a series of benzopyranopyrimidine derivatives was investigated in platelet-dependent and independent pulmonary thromboembolism in mice. Intraperitoneal subacute treatment with 2-morpholino derivative 3c significantly prevented paralysis due to collagen plus epinephrine-induced pulmonary thrombosis while 2-piperidino substituted derivative 3h significantly protected mice from paralysis caused by thrombin-induced intravascular fibrin formation at dosage not affecting bleeding time. These compounds, previously proved to be effective as antiplatelet agents in vitro, were in vivo more potent as antithrombotics than lysine acetylsalicylate and possessed lower prohemorrhagic activity than the reference drug. Although their ineffectiveness on clotting times, PT and APTT, allows the involvement of coagulation pathways to be ruled out, the mechanisms underlying the favourable benefit risk ratio for these two compounds remain to be further clarified. PMID- 14761667 TI - Maternal adrenalectomy affects development of adrenal medulla. AB - This work investigates the effects of maternal adrenalectomy (ADX) on the development of the adrenal medulla. Adrenal catecholamines (AC) were measured at postnatal day (PN) 1, 8, 12 and 22 in rat offspring of ADX dams and in pups of control dams. The pups of ADX rats showed a reduction in AC concentrations in the adrenal medulla at PN 1, 12 and 22, although these were higher than in the pups of sham dams at PN 8. Further, in the pups of control mothers, there was an increase in ACs during the first two weeks of life whereas pups of ADX mothers only showed increases in noradrenaline, dopamine and adrenaline levels at day 8. These results suggest that maternal absence of corticosterone affects the medulla catecholamine content during development. These data support the idea that a maternal glucocorticoids are involved in the differentiation or/and maturation of the adrenal medulla. PMID- 14761668 TI - Nucleic acid synthesis in cancerous cells under the effect of gnidilatimonoein from Daphne mucronata. AB - Cytotoxicity evaluation of gnidilatimonoein, the most active isolated diterpene ester from Daphne mucronata [Sadeghi H, Mianabadi M, Yazdanparast R, (2002) Journal of Tropical. Medicinal Plant1 3: 169-173], revealed the strong antiproliferative activity among several different human cancer cell lines (K562, CCRF-CEM, HL-60 and MOLT-4 leukemia cell lines, LNCaP-FGC-10 a prostate cancer cell line) and a mouse BALB/C fibrosarcoma cell line (WEHI-164). Using flow cytometry technique, it was found that treatment of the most responsive cells (K562) with gnidilatimonoein inhibited the progression of cells through G1 phase by almost 15% compared to the untreated cells. The population of the treated cells in the S and G2 phases also reduced by 8.3% and 5.4%, respectively. Based on the extent of [3H]-thymidine and [3H]-uridine incorporation into DNA and RNA, respectively, the major metabolic effects of gnidilatimonoein were found to be mainly on DNA and to a less extent on RNA synthesis. Additionally, the activity of inosine-5'-monophosphate dehydrogenase (IMPDH), under the effects of genidilatimonoein, was reduced in the treated cells by 44%. These data strongly suggest that the purine biosynthetic pathway is significantly affected by gnidilatimonoein. PMID- 14761669 TI - Combination of metformin and thiazolidindiones restore insulin signalling in insulin-resistant cultured myotubes. AB - We examined the effect of combination of thiazolidinediones (TZDs) and metformin on insulin-resistant skeletal muscle cells. The combined use of TZDs and metformin resulted in maximum tyrosine phosphorylation of insulin receptor (IR) and insulin receptor substrate-1 (IRS-1) at 12.5 microM of TZDs and 100 microM of metformin as compared to the maximum tyrosine phosphorylation of IR and IRS-1 achieved at 50 microM of TZDs or 400 microM of metformin. The glucose uptake was significantly high at the combination of lower concentration (12.5 microM of TZDs and 100 microM of metformin) as compared to the combination of higher concentration (50 microM of TZDs and 400 microM of metformin). Results demonstrated that (1) Additive effect on insulin sensitization can be achieved by a combination of TZDs and metformin at lower concentration; (2) combination of TZDs and metformin act on insulin signaling molecules in insulin resistance; (3) in vitro system has the potentiality to determine possible target molecule(s) and mechanism of action of drugs. PMID- 14761670 TI - The effect of taurine on cholesterol degradation in mice fed a high-cholesterol diet. AB - The hypocholesterolemic effect of taurine was examined in mice fed a high cholesterol diet containing 1% cholesterol and 0.25% sodium cholate. Male C57BL/6 mice were divided into 3 groups: control group (HC), 1% taurine-supplemented group (HCT+), and taurine-deficient group (HCT-) produced by supplying 0.5% guanidinoethyl sulfonate (GES) solution ad libitum instead of water. After they were fed with the respective diet or drinking water for 4 weeks, the liver taurine level was reduced 80% in the HCT- group compared with that in the HC group, although there was no difference in the serum taurine amount between the two groups. The formation ratio of cholesterol gallstones increased from 71% to 100% by taurine deficiency, and decreased to 0% by taurine supplementation. Compared with the HC group, serum and liver cholesterol significantly decreased, and the excretion of fecal bile acid notably rose in the HCT+ group but tended to lower in the HCT- group. There were no differences in LDL receptor protein level among the three groups. In the subsequent experiment, triglycerides (TG) secretion rate was determined and found to be significantly suppressed by taurine supplementation. In conclusion, it is suggested that taurine does not up-regulate LDL receptor protein level, and the decrease in cholesterol in the circulation is mainly due to its suppressive effect on TG secretion from the liver. PMID- 14761671 TI - Differentially expressed genes in hypertensive rats developing cerebral ischemia. AB - The molecular events occurring after cerebral ischemia in hypertension may include de novo expression of numerous genes. Receptor genes are predominantly involved in the process of cell death, neuroprotection and reconstruction after ischemic injury. Ischemic stroke was observed in the non-genetic, non-surgical model of hypertension, the cold-induced hypertensive rat. In hypertensive rats suppression subtractive hybridization analysis was used to identify differentially expressed receptor genes in stroke-tissue compared to normal rat brain. We found 76 genes predominantly expressed in hypertensive rat stroke tissue. These predominantly expressed genes included genes involved in energy metabolism, signal transduction/cell regulation, and replication/transcription/translation. For example, the T3 receptor alpha was predominantly expressed in stroke-tissue, indicating that regeneration of nerves in stroke tissue may be facilitated by increased T3 receptor alpha expression. PMID- 14761672 TI - Monkey corticotropin-releasing factor1 receptor: Complementary DNA cloning and pharmacological characterization. AB - The full-length complementary DNA (cDNA) of monkey corticotropin-releasing factor type 1 (CRF1) receptor was isolated from a rhesus monkey (Macaca mulatta) amygdala cDNA library. The cloned monkey CRF1 receptor cDNA has 2,374 bp with an open reading frame encoding a 415-amino acid protein. The sequence of the monkey CRF1 receptor cDNA showed a high degree of sequence identity with other species of CRF1 receptors, and being 99.5% identical to human CRF1 receptors. When monkey CRF1 was expressed into COS-7 cells, high specific binding of [125I]-ovine CRF was observed. CRF and CRF-related peptides inhibited [125I]-ovine CRF binding in a concentration-dependent manner. IC50 values of ovine CRF, human/rat CRF, sauvagine and urotensin I were 23.5 +/- 7.4, 22.7 +/- 10.8, 27.5 +/- 12.3 and 14.2 +/- 7.0 nM, respectively. CRF1 receptor specific antagonists, such as CP 154,526, SC241 and CRA1000, also inhibited the [125I]-ovine CRF binding, with IC50 values of 3.9 +/- 0.4, 43.5 +/- 8.0 and 19.8 +/- 2.0 nM, respectively. GTP and its nonhydrolyzed analogue, GTPgammaS, reduced [125I]-ovine CRF binding, while ATP had a negligible effect, thereby indicating that the monkey CRF1 receptor belongs to a family of G-protein coupled receptors. CRF and its related peptides increased cyclic AMP formation concentration-dependently in COS-7 cells transiently expressing the monkey CRF1 receptor. Monkey CRF1 was expressed abundantly in the pituitary, cerebral cortex, hippocampus, amygdala and cerebellum. Thus the monkey CRF1 receptor and the human CRF1 receptor have similar molecular and pharmacological characteristics. PMID- 14761673 TI - Postnatal dexamethasone and long term learning and memory functions in developing rats: Effect of postnatal age and gender. AB - In this study, we investigated the effect of dexamethasone on the long-term learning and memory functions in developing rats. In Sprague-Dawley rat pups, we administered a daily dose of dexamethasone (0.5 mg/kg/day) for three consecutive days in three groups of animals: the "ultra-early" group received steroids on postnatal days (PND) 1-3; the "early" group received the drug on PNDs 8-10, and the "late" group received the drug on PNDs 28-30. The control group was not given any medication. All animals underwent structured CNS examinations beginning on PND 15, and continued through PND 20. The pups were tested for spatial learning and memory functions using the Morris Water Maze (MWM) on PNDs 31 through 35, 45 through 49, and 59 through 63. They were also tested for reward-based learning and memory functions using Radial Arm Maze (RAM) on PNDs 70 through 72. We analyzed the effect of dexamethasone, postnatal age, and sex on neurological milestones, and learning and memory functions. We found that neurological examination findings were similar in all groups, as were the results of the reward-based learning using RAM. However, in the MWM, the total distance of swimming and the total time to find the hidden platform showed considerable difference among the groups. Although these functions improved with postnatal age, the female pups in all three steroid groups, and the male pups in the late steroid group lagged significantly in learning and memory functions compared to the controls, and such lags were transient. However, the interaction terms between dexamethasone, age, and sex were also significant in MWM test results. Steroids administered postnatally may have transient, retarding effect on learning and memory functions, and that animal age and sex may modify such effects. Such lags are not global, but specific to the types of memory tests used, implicating different neural circuitries in the pathogenesis of such abnormalities. Although transient, if such adverse effects occur at critical phases during brain maturation, the implications for poor, long-term outcomes may be more significant. The mechanisms underlying such changes need to be explored. PMID- 14761674 TI - Acidic FGF enhances functional regeneration of adult dorsal roots. AB - It has been well documented that the regeneration of sensory axons severed in the dorsal roots into the spinal cord is largely inhibited in adult mammals. We investigated whether peripheral nerve grafts combined with acidic fibroblast growth factor (aFGF) could induce the regeneration of transected dorsal roots in adult rats, as evaluated by cortical somatosensory evoked potentials (SEPs). Median nerve (forelimb) stimuli produced consistent responses in the primary somatosensory cortex of normal rats, but these were completely eliminated after the transection of cervical 6th - 8th roots. The dorsal root stumps were immediately anastomosed to the cord with intercostal nerve grafts. Subsequently, aFGF in fibrin glue was administered to the grafted area. Four to twenty weeks after rhizotomy, six of the seven rats receiving such reconstruction had recovery of SEPs. The reappearing SEPs typically showed similar waveforms and latencies as normal ones. They were eliminated by retransection of the repaired roots, thus verifying their source as the regenerated roots. We present here substantial evidence that aFGF enhances the functional restoration of cut dorsal roots. Cortical SEPs is considered a useful tool in evaluating such regeneration. These results may offer therapeutic potential in the treatment of dorsal root injuries. PMID- 14761675 TI - Crataegus special extract WS 1442 improves cardiac function and reduces infarct size in a rat model of prolonged coronary ischemia and reperfusion. AB - In Germany, hydroalcoholic extracts from hawthorn (Crataegus spp.) leaves with flowers are approved drugs for the treatment of mild forms of heart insufficiency. Besides cardiotonic effects these herbal remedies have been shown to possess cardioprotective properties. We now evaluated if treatment of rats with the Crataegus special extract WS 1442 also improves cardiac function and prevents myocardial infarction during prolonged ischemia and reperfusion lasting for 240 and 15 min, respectively. Oral administration of WS 1442 (10 or 100 mg x kg(-1) x day(-1)) for 7 days before ligation of the left coronary artery dose dependently suppressed the decrease of the pressure rate product. WS 1442 treatment also attenuated the elevation of the ST-segment in the ECG, diminished the incidence of ventricular fibrillations (control: 67%; 10 mg x kg(-1): 64%; 100 mg x kg(-1): 27%) and reduced the mortality rate (control: 47%; 10 mg.kg(-1): 27%; 100 mg x kg(-1): 9%). Furthermore, the area of myocardial infarction within the ischemic zone was significantly smaller in treated rats (10 mg x kg(-1): 64.3 +/- 5.1%; 100 mg x kg(-1): 42.8 +/- 4.1%) when compared with controls (78.4 +/- 2.6%). It is suggested that these pharmacological effects are accounted for by the combined antioxidative, leukocyte elastase inhibiting and endothelial nitric oxide (NO) synthesis enhancing properties of WS 1442. PMID- 14761676 TI - 2,3,7,8-tetrachlorodibenzo-p-dioxin augments the modulation of gene expression mediated by the thyroid hormone receptor. AB - Previously, we reported on genes whose expression was highly modulated by T3 in the HeLaTR cells that stably expressed the thyroid hormone receptor (TR). In this study, we examined the effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on TR-mediated gene expression. In the HeLaTR cells, T3 induced the expression of the reporter gene in a thyroid hormone responsible element (TRE)-dependent manner. When the cells were cultured in the presence of T3, the addition of TCDD but not 4-hydroxy-2',3,4',5,6'-pentachlorobiphenyl (PCB-OH), bisphenol A (BPA), or di(2-ethylhexyl)phthalate (DEHP) to the culture media further enhanced the T3 induced expression of the reporter gene. RT-PCR revealed that mRNA levels of 4 1BB, fmfc, PSCA, PSG7, RANTES, and TRAF1, which were highly increased by T3, were further elevated in cells exposed to T3 and TCDD. Also, the mRNA level of BMP6, which was decreased by T3, further declined in the cells exposed to both T3 and TCDD. In contrast to the effect of TCDD, PCB-OH suppressed the modulation of these gene expressions by T3. Neither TCDD nor PCB-OH alone affected the expression of 4-1BB, fmfc, PSCA, PSG7, RANTES, TRAF1, or BMP6. These results indicate that TCDD augments the cellular responses to T3 by hyperactivating TR mediated gene expression, whereas PCB-OH suppresses cellular responses to T3 by negatively regulating it. Based on these findings, enzyme-linked immunosorbent assay (ELISA) for the PSCA protein in the HeLaTR cells was established. Such assays will be useful to monitor the effects of endocrine disrupting chemicals (EDCs) on TR-mediated gene expression. PMID- 14761677 TI - Postnatal remodeling of the neural components of the epithelial-mesenchymal trophic unit in the proximal airways of infant rhesus monkeys exposed to ozone and allergen. AB - Nerves and neuroendocrine cells located within the airway epithelium are ideally situated to sample a changing airway environment, to transmit that information to the central nervous system, and to promote trophic interactions between epithelial and mesenchymal cellular and acellular components. We tested the hypothesis that the environmental stresses of ozone (O(3)) and house dust mite allergen (HDMA) in atopic infant rhesus monkeys alter the distribution of airway nerves. Midlevel bronchi and bronchioles from 6-month-old infant monkeys that inhaled filtered air (FA), house dust mite allergen HDMA, O(3), or HDMA + O(3) for 11 episodes (5 days each, 0.5 ppm O(3), 8 h/day followed by 9 days recovery) were examined using immunohistochemistry for the presence of Protein gene product 9.5 (PGP 9.5), a nonspecific neural indicator, and calcitonin gene-related peptide (CGRP). Along the axial path between the sixth and the seventh intrapulmonary airway generations, there were small significant (P < 0.05) decrements in the density of epithelial nerves in monkeys exposed to HDMA or O(3), while in monkeys exposed to HDMA + O(3) there was a greater significant (P < 0.05) reduction in epithelial innervation. In animals exposed to O(3) or HDMA + O(3) there was a significant increase in the number of PGP 9.5 positive/CGRP negative cells that were anchored to the basal lamina and emitted projections in primarily the lateral plain and often intertwined with projections and cell bodies of other similar cells. We conclude that repeated cycles of acute injury and repair associated with the episodic pattern of ozone and allergen exposure alter the normal development of neural innervation of the epithelial compartment and the appearance of a new population of undefined PGP 9.5 positive cells within the epithelium. PMID- 14761678 TI - A ginseng saponin metabolite-induced apoptosis in HepG2 cells involves a mitochondria-mediated pathway and its downstream caspase-8 activation and Bid cleavage. AB - 20-O-(beta-D-glucopyranosyl)-20(S)-protopanaxadiol (IH901), an intestinal bacterial metabolite of ginseng saponin formed from ginsenosides Rb1, Rb2, and Rc, is suggested to be a potential chemopreventive agent. Here, we show that IH901 induces apoptosis in human hepatoblastoma HepG2 cells. IH901 led to an early activation of procaspase-3 (12 h posttreatment), and the activation of caspase-8 became evident only later (18 h posttreatment). Caspase activation was a necessary requirement for apoptosis because caspase inhibitors significantly inhibited cell death by IH901. Treatment of HepG2 cells with IH901 also induced the cleavage of cytosolic factors such as Bid and Bax and translocation of truncated Bid (tBid) to mitochondria. A time-dependent release of cytochrome c from mitochondria was observed, which was accompanied by activation of caspase-9. A broad-spectrum caspase inhibitor, N-benzyloxycarbonyl-Val-Ala-Asp fluoromethylketone (zVAD-fmk), and a specific inhibitor for caspase-8, N benzyloxycarbonyl-Ile-Glu-Thr-Asp-fluoromethylketone (zIETD-fmk), abrogated Bid processing and translocation, and caspase-3 activation. Cytochrome c release was inhibited by zVAD-fmk, however, the inhibition by zIETD-fmk was not complete. The activation of caspase-8 was inhibited not only by zIETD-fmk but also by zVAD-fmk. The results, together with the kinetic change of caspase activation, indicate that activation of caspase-8 occurred downstream of caspase-3 and -9. Our data suggest that the activation of caspase-8 after early caspase-3 activation might act as an amplification loop necessary for successful apoptosis. Primary hepatocytes isolated from normal Sprague-Dawley rats were not affected by IH901 (0-60 microM). The very low toxicity in normal hepatocytes and high activity in hepatoblastoma HepG2 cells suggest that IH901 is a promising experimental cancer chemopreventive agent. PMID- 14761679 TI - In vivo erythroid recovery following paclitaxel injury: correlation between GATA 1, c-MYB, NF-E2, Epo receptor expressions, and apoptosis. AB - Paclitaxel (Px) is a cancer chemotherapeutic agent that causes bone marrow (BM) cytotoxicity by microtubule stabilization and by modifications in the expression of several genes. Hematopoietic progenitors show severe alterations following Px injury. Erythropoietic recovery should be accompanied by changes in the expression of transcription factors such as c-MYB, GATA-1, NF-E2, Bcl-x(L), and erythropoietin receptor (Epo-R). The aim of this work was to study the in vivo recovery of erythropoiesis and to correlate transcription factors, Bcl-x(L), and Epo-R expressions to apoptosis and changes in proliferation of murine erythroid progenitors following a single dose of Px (29 mg/kg, i.p.). BM total and differential cellularities, apoptosis (TdT-mediated dUTP Nick-End Labeling [TUNEL] assay), clonogenic assays, and immunoblots for transcription factors, Epo R, and Bcl-x(L) were performed each day for 5 days post-injury. Apoptosis (24 +/- 0.81%, P < 0.01), inhibition of colony growth (burst-forming units-erythroid [BFU E] and granulocyte-erythroid-macrophage [GEM]), and decrease in BM cellularities (28 +/- 4.2% of control) were maximal at 24 h following Px. The highest apoptosis was concomitant with the lowest BM cellularities. Apoptosis returned to normal values (3.08 +/- 0.61%) by day 3 post-Px. Up-regulation of c-MYB, GATA-1, Epo-R, and Bcl-x(L) expressions were observed between 24 and 48 h following Px. Correlations among c-MYB, GATA-1, Bcl-x(L), and Epo-R were extremely significant. Maximal expression of NF-E2 was observed on day 3 concomitant with the rise (threefold) of early erythroid precursors (BFU-E). Thus, cells that survive injury seem to be stimulated to produce early (24-48 h) erythroid-related and antiapoptotic proteins. Therefore, the results suggest an in vivo interplay between specific transcription factors and Bcl-x(L) during progenitor cell survival and proliferation; mechanisms triggered to restore size and composition of the erythroid compartment. PMID- 14761680 TI - Protective effects of ursodeoxycholic acid against 2,3,7,8-tetrachlorodibenzo-p dioxin-induced testicular damage in mice. AB - The protective effect of ursodeoxycholic acid (UDCA), a biliary component found in bears, on 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)-induced testicular damage in mice was investigated. Fifty C57BL/6J mice were equally divided into five groups. The mice in the control group received the vehicle and standard chow. The single TCDD treatment group received 27.5 microg/kg of TCDD subcutaneously. The UDCA-included treatment group received pulverized chow containing 0.125%, 0.25% and 0.5% UDCA, respectively, for 70 days starting 10 days before TCDD injections. The body and testicular weights were shown to be decreased in the single TCDD treatment group, while the decrease was prevented by UDCA added to the chow. In addition, the decrease in the serum-luteinizing hormone (LH) or the follicle stimulating hormone (FSH) secondary to a TCDD injection was not observed in the UDCA-included treatment group. Contrary to the single TCDD treatment group, the germinal epithelium and intercellular space were relatively well preserved in the UDCA-included treatment group. Adding UDCA also normalized TCDD-induced irregular ultrastructural changes such as development of phagolysosomes, inflated smooth endoplasmic reticulum (SER), dilated and altered mitochondria, necrosis and completely damaged seminiferous tubules. Moreover, in the experiment for Arnt expression, UDCA added to the chow suppressed the TCDD-induced relocation of Arnt from the cytoplasm to the nuclei. In conclusion, TCDD-induced testicular toxicity was effectively protected by UDCA. There was almost complete recovery of the testes in the UDCA-included treatment group. Thus, UDCA may be useful for the prevention and treatment of TCDD-induced testicular damage. PMID- 14761681 TI - The nematode Caenorhabditis elegans as a model of organophosphate-induced mammalian neurotoxicity. AB - Fifteen organic phosphate pesticides were tested by computer tracking for their acute behavioral toxicity with the nematode Caenorhabditis elegans. Thirteen of these 15 chemicals are used as insecticides and are anticholesterase agents. The other two chemicals are used as herbicides. EC50 values for each chemical were compared to the corresponding LD50 acute lethality value in rats and mice. Order of toxicity was found to be significantly correlated in comparisons of C. elegans to both rats and mice. Mechanistic investigations were conducted by assaying 8 of the 15 chemicals for anticholinesterase activity in C. elegans. Significant cholinesterase inhibition was confirmed for five chemicals that had displayed high behavioral toxicity, while three chemicals of low behavioral toxicity showed no significant decrease in cholinesterase activity. Toxicity for two chemicals that do not inhibit cholinesterase in mammals was linked to pH effects. Detailed comparison of individual chemicals and metabolic issues are discussed. These results have positive implications for the use of C. elegans as a mammalian neurological model and support the use of C. elegans in early rounds of chemical toxicity screening. PMID- 14761682 TI - Molecular multiple endpoint embryonic stem cell test--a possible approach to test for the teratogenic potential of compounds. AB - The embryonic stem cell test (EST) examines the cytotoxicity of chemical compounds on embryonic stem (ES) cells and 3T3.A31 fibroblasts. Additionally, the EST measures the ability of ES cells to differentiate into contracting cardiomyocytes following drug exposure. In this study, we introduce new endpoints to obtain a molecular multiple endpoint EST (mme-EST), enabling the identification of potential chemical effects on osteogenic, chondrogenic and neural differentiation in addition to the traditional endpoint of cardiomyocyte differentiation. Six compounds in three classes with known teratogenic in vivo potential were assayed with the mme-EST in a pilot study: penicillin G (non teratogenic), 5-fluorouracil and retinoic acid (strongly teratogenic), diphenylhydantoin, valproic acid and thalidomide (moderately teratogenic). While the traditional EST measures a morphological endpoint, we included molecular markers of differentiation as endpoints. With the mme-EST, every compound could be classified correctly according to its known teratogenic potential in vivo. Penicillin G, 5-fluorouracil and diphenylhydantoin inhibited differentiation of all endpoints equally. Interestingly, valproic acid showed the strongest inhibition of neural differentiation, while thalidomide specifically inhibited osteogenic development. Retinoic acid, on the other hand, supported neural but inhibited chondrogenic and osteogenic differentiation concentration-dependently. Valproic acid and thalidomide, classified incorrectly with the established EST model, were classified correctly with the mme-EST according to their effects on specific endpoints. This pilot study indicates that the predictive value of the EST may be enhanced by including further differentiation endpoints. PMID- 14761683 TI - Relevance of the palatal protein kinase A pathway to the pathogenesis of cleft palate by secalonic acid D in mice. AB - Secalonic acid-D (SAD) is a teratogenic mycotoxin inducing cleft palate (CP) in the offspring of the exposed mice by reducing palatal shelf size secondary to reduced proliferation of the palatal mesenchymal (PM) cells. Co-administration of dimethylsulfoxide (DMSO) reversed the CP-inducing effect of SAD. Although SAD has been shown to affect both protein kinases A (PKA) and C (PKC) pathways, the relevance of each of these pathways to its CP induction is unknown. The present studies were designed to test the hypothesis that the protective effect of DMSO is mediated by its specific reversal of the effect(s) of SAD on one of these two pathways using ELISA-based activity assays, Western blot analysis, electrophoretic mobility shift assays (EMSA), and murine embryonic PM (MEPM) cell growth in culture. Within the PKA pathway, SAD inhibited the activity of the catalytic subunit of PKA and its migration into the nucleus, elevated phosphorylated cyclic AMP (cAMP) response element (CRE)-binding protein (pCREB) level, and reduced the binding of CREB to CRE. In the PKC pathway, SAD reduced the activity of PKC and the binding of transcription factors (TF) to 12-O tetradecanoate-13 phorbol acetate-response element (TRE). SAD also inhibited MEPM cell growth and the expression of the CRE- and TRE-containing gene, proliferating cell nuclear antigen (PCNA). Reversal, by DMSO, of the effects of SAD on MEPM cell growth, on PCNA expression and on all components of the PKA, but not of PKC, pathway suggests that the perturbation of the PKA pathway by SAD is relevant to its induction of CP in mice. PMID- 14761685 TI - Physiological role of mGSTA4-4, a glutathione S-transferase metabolizing 4 hydroxynonenal: generation and analysis of mGsta4 null mouse. AB - The lipid peroxidation product 4-hydroxynon-2-enal (4-HNE) is a strong electrophile that forms covalent adducts with proteins and, to a lesser extent, nucleic acids and phospholipids. The generation of 4-HNE appears to be an inevitable consequence of aerobic metabolism. The metabolism of 4-HNE is mainly, although not entirely, conjugative, and proceeds via Michael addition of glutathione to the double bond of 4-HNE. This reaction is catalyzed by specialized glutathione S-transferases (GSTs) exemplified by the murine mGSTA4-4. To study the (patho)physiological effects of 4-HNE in an intact organism, we disrupted the mGsta4 gene in the mouse. The resulting mGsta4 null mouse expressed no mGsta4 mRNA and no corresponding protein, had a reduced ability to conjugate 4 HNE, and had an increased steady-state level of this aldehyde in tissues. The residual conjugating activity for 4-HNE (23-64% depending on the tissue) is probably attributable to isoforms of glutathione S-transferases which have low catalytic efficiency for 4-HNE but are more abundant than mGSTA4-4, or are upregulated upon mGsta4 gene disruption. Mice homozygous for the disrupted mGsta4 allele were viable and appeared normal except for lower litter size, higher fat content in bones, and greater susceptibility to bacterial infection. The null mice had a significantly lower survival time than wild-type controls when chronically treated with relatively low doses of paraquat, a finding consistent with a role of mGSTA4-4 in the defense against oxidative stress. The mouse model should be useful for the study of degenerative conditions in which 4-HNE is postulated to be a contributing factor. PMID- 14761684 TI - Neuronal nitric oxide synthase and N-methyl-D-aspartate neurons in experimental carbon monoxide poisoning. AB - We measured changes in nitric oxide (NO) concentration in the cerebral cortex during experimental carbon monoxide (CO) poisoning and assessed the role for N methyl-d-aspartate receptors (NMDARs), a glutamate receptor subtype, with progression of CO-mediated oxidative stress. Using microelectrodes, NO concentration was found to nearly double to 280 nM due to CO exposure, and elevations in cerebral blood flow, monitored as laser Doppler flow (LDF), were found to loosely correlate with NO concentration. Neuronal nitric oxide synthase (nNOS) activity was the cause of the NO elevation based on the effects of specific NOS inhibitors and observations in nNOS knockout mice. Activation of nNOS was inhibited by the NMDARs inhibitor, MK 801, and by the calcium channel blocker, nimodipine, thus demonstrating a link to excitatory amino acids. Cortical cyclic GMP concentration was increased due to CO poisoning and shown to be related to NO, versus CO, mediated guanylate cyclase activation. Elevations of NO were inhibited when rats were infused with superoxide dismutase and in rats depleted of platelets or neutrophils. When injected with MK 801 or 7 nitroindazole, a selective nNOS inhibitor, rats did not exhibit CO-mediated nitrotyrosine formation, myeloperoxidase (MPO) elevation (indicative of neutrophil sequestration), or impaired learning. Similarly, whereas CO-poisoned wild-type mice exhibited elevations in nitrotyrosine and myeloperoxidase, these changes did not occur in nNOS knockout mice. We conclude that CO exposure initiates perivascular processes including oxidative stress that triggers activation of NMDA neuronal nNOS, and these events are necessary for the progression of CO-mediated neuropathology. PMID- 14761686 TI - Thyroid hormone, glucocorticoids, and prolactin at the nexus of physiology, reproduction, and toxicology. AB - A symposium at the 2003 Annual Meeting of the Society of Toxicology brought together an expert group of endocrinologists to review how non-reproductive hormones can affect the endocrine system. This publication captures the essence of those presentations. Paul Cooke and Denise Holsberger recapitulate the evidence of how thyroid hormones affect male and female reproduction, and reproductive development. Ray Witorsch summarizes the many effects of glucocorticoids on the reproductive system. Finally, Paul Sylvester reviews the mechanism of action of prolactin, and reminds us that this ancient hormone has many functions beyond lactation. PMID- 14761687 TI - Recombinant luminescent bacterial sensors for the measurement of bioavailability of cadmium and lead in soils polluted by metal smelters. AB - Environmental hazard of heavy metals in soils depends to a large extent on their bioavailability. The approach used in this study enables the determination of bioavailable metals in solid-phase samples. Two recombinant bacterial sensors, one responding specifically to cadmium and the other to lead and cadmium by increase of luminescence (firefly luciferase was used as a reporter) were used to determine the bioavailability of these metals in soil-water suspensions (a contact assay) and respective particle-free extracts. Fifty agricultural soils sampled near zinc and lead smelters in the Northern France containing up to (mg/kg) 20.1 of Cd, 1050 of Pb and 1390 of Zn were analysed. As the soil matrix interferes with the assay, recombinant luminescent control bacteria lacking the metal recognizing protein and corresponding promoter (thus, being not metal inducible) but otherwise comparable to the sensor bacteria (the same host bacterium and plasmid encoding luciferase) were used in parallel to take into account the possible quenching and/or stimulating effects of the sample on the luminescence of the sensor bacteria. Both, chemical and sensor analysis showed that only microg/l levels of metals were extracted from the soil into the water phase (0.1% of the total Cd, 0.07% of Pb and 0.5% of Zn). However, 115-fold more Cd and 40-fold more Pb proved bioavailable if the sensor bacteria were incubated in soil suspensions (i.e., in the contact assay). The bioavailability of metals in different soils varied (depending probably on soil type) ranging from 0.5% to 56% for cadmium and from 0.2% to 8.6% for lead. PMID- 14761689 TI - Effects of chronic exposure of 2,4-dichlorophenol on the antioxidant system in liver of freshwater fish Carassius auratus. AB - There were few reports on the antioxidant response of aquatic organisms exposed to 2,4-dichlorophenol (2,4-DCP). This research explored the hepatic antioxidant responses of fish to long-term exposure of 2,4-DCP for the first time. Freshwater fish Carassius auratus were chosen as experimental animals. The fish were exposed to six different concentrations of 2,4-DCP (0.005-1.0 mg/l) for 40 days and then liver tissues were separated for determination. As shown from the results, 40 days afterwards, the activities of catalase (CAT) and selenium-dependent glutathione peroxidase (Se-GPx) and the content of oxidized glutathione (GSSG) were induced significantly on the whole compared to control group; superoxide dismutase (SOD) responded to 2,4-DCP exposure at only 0.005 mg/l; the content of reduced glutathione (GSH) was suppressed continuously except Group 7; the activity of glutathione reductase was inhibited initially and then restored to control level from Group 4 on; glutathione S-transferase had only slight responses in Groups 3 and 4. Total glutathione (tGSH) and GSH/GSSG ratio were also calculated to analyze the occurrence of oxidative stress. Besides, good dose effect relations, which cover most of the exposure concentration range, were found between 2,4-DCP level and CAT activity, GSSG content, Se-GPx activity, respectively. In conclusion, SOD and Se-GPx may be potential early biomarkers of 2,4-DCP contamination in aquatic ecosystems, and further studies will be necessary. PMID- 14761688 TI - Calculating pesticide sorption coefficients (Kd) using selected soil properties. AB - Pesticide soil/solution distribution coefficients ( Kd values), commonly referred to as pesticide soil sorption values, are utilized in computer and decision aid models to predict soil mobility of the compounds. The values are specific for a given chemical in a given soil sample, normally taken from surface soil, a selected soil horizon, or at a specific soil depth, and are normally related to selected soil properties. Pesticide databases provide Kd values for each chemical, but the values vary widely depending on the soil sample on which the chemicals were tested. We have correlated Kd values reported in the literature with the reported soil properties for an assortment of pesticides in an attempt to improve the accuracy of a Kd value for a specific chemical in a soil with known soil properties. Mathematical equations were developed from regression equations for the related properties. Soil properties that were correlated included organic matter content, clay mineral content, and/or soil pH, depending on the chemical properties of the pesticide. Pesticide families for which Kd equations were developed for 57 pesticides include the following: Carboxy acid, amino sulfonyl acid, hydroxy acid, weakly basic compounds and nonionizable amide/anilide, carbamate, dinitroaniline, organochlorine, organophosphate, and phenylurea compounds. Mean Kd values for 32 additional pesticides, many of which had Kd values that were correlated with specific soil properties but for which no significant Kd equations could be developed are also included. PMID- 14761690 TI - Distribution pattern of ambient cadmium in wetland ponds distributed along an industrial complex. AB - Water and sediment samples collected from 18 wetland ponds within and outside industrial areas were examined for cadmium concentration and water quality parameters during the period of January to July 1996. The Cd contents in gill, liver, mantle and shell of freshwater mussel (Lamellidens marginalis) as well as leaves and roots of water hyacinth Eichhornia those occurred in these ponds were also estimated. Cd concentration ranged from 0.006 to 0.7025 mg/l in water and from 7 to 77 microg/gdw in sediments of all the ponds investigated. The amount of Cd occurring in water and sediment was much higher in concentrations in the ponds located in Captain Bheri and Mudiali farm close to industrial areas, compared to remaining ponds located outside the industrial belt. Lamellidens marginalis procured from Mudiali and Captain Bheri ponds showed regardless of size, tissue and season of collection significantly higher Cd concentration than did those from other ponds. Likewise, tissue Cd in Eichhornia collected from Mudiali pond was as high as 125-152 microg/gdw in root and 21-63 microg/gdw in leaves compared to 40-108 microg/gdw in root and 9-43 microg/gdw in leaves in the remaining ponds. Seasonal variability of Cd was clear-cut; the concentration was relatively higher in water and sediment in all ponds during summer than during monsoon season or winter. Size-wise, smaller groups showed the highest concentrations of Cd in all tissues of Lamellidens compared with medium and large size groups. Concentration factor for all tissues of Lamellidens regardless of size and season, was inversely proportional with the ambient Cd concentrations. Concentration factor estimated for all tissues in all ponds and all seasons was in the order: liver>gill>shell>mantle. As all ponds located outside the industrial belt showed Cd concentrations ranging from 0.006 to 0.049 mg/l, it is suggested that these wetlands do not pose serious risk to the environment. PMID- 14761691 TI - Bisphenol A induces yolk-sac oedema and other adverse effects in landlocked salmon (Salmo salar m. sebago) yolk-sac fry. AB - Accumulation and toxicity of waterborne bisphenol A were studied in landlocked salmon (Salmo salar m. sebago) yolk-sac fry. In a short-term (96 h) exposure to five bisphenol A concentrations yolk-sac fry had higher accumulation rates and bioconcentration factors (BCF96) than earlier studies have shown for salmon eggs. Furthermore, the conditional uptake rate constant tended to decrease as exposure concentration increased. Fry were also exposed to bisphenol A for 42 days at three concentrations (10, 100 and 1000 microg/l), and changes in behaviour, morphology and histological structure were observed. After 6 days of exposure, the highest concentration (1000 microg/l) of bisphenol A caused fluid accumulation (oedema) in the yolk sac and haemorrhages in the front part of the yolk sac and in the head around the gill arches. Later on, the fry at 1000 microg/l showed phlegmatic behaviour and had darker skin coloration than the fry in the other treatments. At the two highest concentrations (100 and 1000 microg/l) histological changes were seen in liver cell nuclei, where strongly stained fragments were observed. In the control fry and the fry exposed to 10 microg/l the nucleolus was clearly visible and spherical in shape and no strongly stained fragments were present. This study shows that high concentrations of bisphenol A may have both morphological and histological effects on salmon yolk sac fry. PMID- 14761692 TI - Total bacterial and fungal population after chlorpyrifos and quinalphos treatments in groundnut (Arachis hypogaea L.) soil. AB - Short-term inhibitory effect on the total bacterial population was observed after chlorpyrifos and quinalphos applications in the groundnut fields, which recovered within 60 days after seed treatment and by 45 days of soil treatment. The fungal population was significantly enhanced after chlorpyrifos treatment whereas quinalphos inhibited the fungal population during the initial days of treatment but no effect was observed after 60 days of treatment. The residues of chlorpyrifos and quinalphos in the treated soil were not persistent and their half-lives ranged from 7.0 to 9.2 days and 13.2 to 20.6 days, respectively. PMID- 14761693 TI - Treatment and detoxification of a sanitary landfill leachate. AB - The leachate from an old sanitary landfill (Gramacho Metropolitan Landfill, Rio de Janeiro) was characterized and submitted to coagulation and flocculation treatment followed by ozonation and ammonia stripping. The performance of the treatment was assessed by monitoring the removal of organic matter (COD and TOC), ammonium nitrogen and metals. Detoxification was assessed by determining acute toxicity, using the following organisms: Vibrio fisheri, Daphnia similis, Artemia salina and Brachydanio rerio. Membrane fractionation was employed to infer the range of molecular masses of the pollutants found in the effluent, as well as the toxicity associated to these fractions. Of the techniques under investigation, coagulation and flocculation followed by ammonia stripping were the most effective for toxicity and ammonium nitrogen removal. Membrane fractionation was effective for COD removal; however, acute toxicity was almost the same in all the fractionated samples. Ozonation was moderately effective for COD removal, but significant toxicity removal was only attained when high ozone doses were used. PMID- 14761694 TI - Adsorption of sugar beet herbicides to Finnish soils. AB - Three sugar beet herbicides, ethofumesate, phenmedipham and metamitron, are currently used on conventional sugar beet cultivation, while new varieties of herbicide resistant (HR) sugar beet, tolerant of glyphosate or glufosinate ammonium, are under field testing in Finland. Little knowledge has so far been available on the adsorption of these herbicides to Finnish soils. The adsorption of these five herbicides was studied using the batch equilibrium method in 21 soil samples collected from different depths. Soil properties like organic carbon content, texture, pH and partly the phosphorus and oxide content of the soils were tested against the adsorption coefficients of the herbicides. In general, the herbicides studied could be arranged according to their adsorption coefficients as follows: glyphosate>phenmedipham>ethofumesate approximately glufosinate-ammonium>metamitron, metamitron meaning the highest risk of leaching. None of the measured soil parameters could alone explain the adsorption mechanism of these five herbicides. The results can be used in model assessments of risk for leaching to ground water resulting from weed control of sugar beet in Finland. PMID- 14761695 TI - Impact assessment of treated/untreated wastewater toxicants discharged by sewage treatment plants on health, agricultural, and environmental quality in the wastewater disposal area. AB - Studies were undertaken to assess the impact of wastewater/sludge disposal (metals and pesticides) from sewage treatment plants (STPs) in Jajmau, Kanpur (5 MLD) and Dinapur, Varanasi (80 MLD), on health, agriculture and environmental quality in the receiving/application areas around Kanpur and Varanasi in Uttar Pradesh, India. The raw, treated and mixed treated urban wastewater samples were collected from the inlet and outlet points of the plants during peak (morning and evening) and non-peak (noon) hours. The impact of the treated wastewater toxicants (metals and pesticides) on the environmental quality of the disposal area was assessed in terms of their levels in different media samples viz., water, soil, crops, vegetation, and food grains. The data generated show elevated levels of metals and pesticides in all the environmental media, suggesting a definite adverse impact on the environmental quality of the disposal area. The critical levels of the heavy metals in the soil for agricultural crops are found to be much higher than those observed in the study areas receiving no effluents. The sludge from the STPs has both positive and negative impacts on agriculture as it is loaded with high levels of toxic heavy metals and pesticides, but also enriched with several useful ingredients such as N, P, and K providing fertilizer values. The sludge studied had cadmium, chromium and nickel levels above tolerable levels as prescribed for agricultural and lands application. Bio monitoring of the metals and pesticides levels in the human blood and urine of the different population groups under study areas was undertaken. All the different approaches indicated a considerable risk and impact of heavy metals and pesticides on human health in the exposed areas receiving the wastewater from the STPs. PMID- 14761696 TI - Studies on lethal concentrations and toxicity stress of some xenobiotics on aquatic organisms. AB - Three widely used xenobiotics pentachlorophenol (PCP), 2,4-dichlorophenoxyacetic acid (2,4-D) and 2-chloro-2,6-diethyl-N-(butoxymethyl) acetanilide (Butachlor) are evaluated for acute toxicity and stress behavior on freshwater fish (Heteropneustes fossilis, Clarias batrachus, Channa punctatus) and mosquito larvae (Culex pipiens fatigans). The experiment was carried out by medium treatment using intermittent flow-through system. Median lethal concentrations (LC50) were calculated by probit analysis. The LC50 values and 95% confidence intervals showed variable range for tested chemicals. Mosquito larvae generally appeared resistant than fish, while H. fossilis was found to be most sensitive. Stress signs in the form of behavioral changes are also observed. Both types of organisms are recommended as good bioindicator for the risk assessment of aquatic environment due to chemicals tested. PMID- 14761697 TI - Organic pollutant burden of the giant mussels Choromytilus chorus from the south central Chilean coast. AB - A comparative quantitative analysis was made on the contents of organochlorines (OCh) and polycyclic aromatic hydrocarbons (PAH) in tissues of the giant mussel Choromytilus chorus from three bays in south-central Chile exposed to various degrees of pollutant input. Mussels from the three bays contained levels of OCh near the detection limits of method, with a tendency to be slightly higher in the polluted bay (San Vicente) than in the other bays (Corral and Yaldad). PAH levels and types in Corral and Yaldad were well below of carcinogenicity permitted in mussels for human consumption. PMID- 14761698 TI - Effects of pH on the mortality and accumulation of copper in tissues of Oreochromis niloticus. AB - In the present study, effect of pH on the mortality and accumulation of copper in various tissue and organs of Oreochromis niloticus were tested at varying concentrations of copper in the medium and over different periods of time. Experimental animals were exposed to pH 5.5, 7.8 and 9.5 and 0.1, 0.5, 1.0 and 5.0 ppm copper over periods of 7, 15 and 30 days in liver, gills and muscle were determined using atomic absorption spectrophotometric techniques. The rate of mortality at 1.0 and 5.0 ppm Cu was 100% after 7 days of exposure at pH 5.5 while at 5.0 ppm Cu was 66% after 30 days of exposure at pH 7.8. No mortality was observed in any of the copper concentrations tested at pH 9.5. In all pH levels, tissue accumulation of copper increased with increasing concentrations of copper in the medium at a given exposure period. In all pH values tested, highest levels of copper were found in the liver of O. niloticus, followed by the gills and muscle tissues. Accumulation of copper in all tissues were higher at pH 5.5 compared with the other pH values in all the conditions tested. PMID- 14761699 TI - Recovery of acetylcholine esterase activity of Drawida willsi (Oligochaeta) following application of three pesticides to soil. AB - The recovery of acetylcholine esterase (AChE) activity of a dominant crop field earthworm (Drawida willsi, Michaelsen) was investigated under laboratory conditions following the application of two recommended agricultural (single and double) doses of butachlor (1.1 and 2.2 mga.i.kg(-1) dry soil), malathion (2.2 and 4.4 mga.i.kg(-1) dry soil) and carbofuran (1.1 and 2.2 mga.i.kg(-1) dry soil) to the soil. A sharp decline in the AChE activity of D. willsi was observed up to 9 and 12 days following treatment of carbofuran and malathion in both single and double doses, respectively, whereas very little inhibition was noticed in case of butachlor. D. willsi worms took 45 and 75 days to resume normal AChE activity after exposure to both single and double doses of malathion and carbofuran, respectively. Earlier [Soil Biol. Biochem. 31 (1999) 363-366], [Ph.D. thesis, Sambalpur University, Orissa, India, 2003] and [Pedobiologia (spl. issue), in press] reported that D. willsi takes 75-90 days and 90-105 days to resume normal growth and reproduction following application of both single and double agricultural doses of malathion and carbofuran, respectively. On the basis of the present and previous studies, we strongly suggest that the time gap between the first and second application of malathion, irrespective of single and double dosage, should be at least 90 days, whereas it should be at least 105 days for carbofuran. Butachlor was found to be very toxic, suppressing growth, sexual maturation and cocoon production of D. willsi at both single and double doses [Ph.D. thesis, Sambalpur University, Orissa, India, 2003]. We therefore suggest that application of organochlorine pesticides like butachlor should be avoided as far as possible to ensure maintenance of good soil health. PMID- 14761701 TI - The role of pregnane neurosteroids in ethanol withdrawal: behavioral genetic approaches. AB - Within the last 20 years, rapid nongenomic actions of steroid hormones have been demonstrated to occur via an interaction with ligand-gated ion channels. For example, the pregnane neurosteroid allopregnanolone (ALLOP) is a potent positive modulator of gamma-aminobutyric acid(A) (GABA(A)) receptors. The physiological significance of fluctuations in endogenous ALLOP levels has been investigated with regard to disease states and the effect of therapeutic agents on ALLOP levels. Because the pharmacological profile of ALLOP is similar to that of ethanol (EtOH), the modulatory effect of pregnane neurosteroids on EtOH dependence and withdrawal will be the focus of this review. Data on the effects of chronic EtOH exposure and withdrawal on pregnane neurosteroid levels, biosynthetic enzymes, and changes in neurosteroid sensitivity will be summarized. Results from genetic animal models indicate that seizure-prone animals have a persistent decrease in endogenous ALLOP levels during EtOH withdrawal in conjunction with tolerance to ALLOP's anticonvulsant effect. Manipulation of endogenous ALLOP levels with finasteride also markedly reduced the severity of chronic EtOH withdrawal. Gene mapping studies provide a hint for an interaction between genes for GABA(A) receptor subunits and the biosynthetic enzyme 5alpha reductase. Overall, the results are suggestive of a relationship between endogenous pregnane neurosteroid levels and behavioral changes in excitability during EtOH withdrawal, consistent with recent findings in humans. While the findings with ALLOP emphasize the therapeutic potential of neurosteroid treatment during EtOH withdrawal, the gene mapping studies suggest that pregnane neurosteroid biosynthesis may represent a target for therapeutic intervention in the treatment of alcohol dependence. PMID- 14761702 TI - Autocrine and paracrine actions of natriuretic peptides in the heart. AB - The natriuretic peptides, atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP), and C-type natriuretic peptide (CNP), are a family of polypeptide mediators exerting numerous actions in cardiovascular homeostasis. ANP and BNP are cardiac derived, being secreted and up-regulated in myocardium in response to many pathophysiological stimuli. CNP is an endothelium-derived mediator. The classical endocrine effects of ANP and BNP on fluid homeostasis and blood pressure, especially in conditions characterised by left ventricular dysfunction, are well recognised and extensively researched. However, there is accumulating evidence that, in addition to endocrine actions, ANP and BNP exhibit important autocrine and paracrine functions within the heart and coronary circulation. These include regulation of myocyte growth, inhibition of fibroblast proliferation and extracellular matrix deposition, a cytoprotective anti ischaemic (preconditioning-like) function, and influences on coronary endothelium and vascular smooth muscle proliferation and contractility. Most if not all of these actions can be ascribed to particulate guanylyl cyclase activation because the ANP/BNP receptor, natriuretic peptide receptor (NPR)-A, has an intracellular guanylyl cyclase domain. Subsequent elevation of the intracellular second messenger cGMP may exert diverse physiological effects through activation of cGMP dependent protein kinases (cGK), predominantly cGK-I. However, there appear to be other contributory mechanisms in several of these actions, including the augmentation of nitric oxide synthesis. These diverse actions may represent counterregulatory mechanisms in the pathophysiology of many cardiovascular diseases, not just those typified by left ventricular dysfunction. Ultimately, insights from the autocrine/paracrine actions of natriuretic peptides may provide routes to therapeutic application in cardiac diseases of natriuretic peptides and drugs that modify their availability. PMID- 14761703 TI - Hallucinogens. AB - Hallucinogens (psychedelics) are psychoactive substances that powerfully alter perception, mood, and a host of cognitive processes. They are considered physiologically safe and do not produce dependence or addiction. Their origin predates written history, and they were employed by early cultures in a variety of sociocultural and ritual contexts. In the 1950s, after the virtually contemporaneous discovery of both serotonin (5-HT) and lysergic acid diethylamide (LSD-25), early brain research focused intensely on the possibility that LSD or other hallucinogens had a serotonergic basis of action and reinforced the idea that 5-HT was an important neurotransmitter in brain. These ideas were eventually proven, and today it is believed that hallucinogens stimulate 5-HT(2A) receptors, especially those expressed on neocortical pyramidal cells. Activation of 5-HT(2A) receptors also leads to increased cortical glutamate levels presumably by a presynaptic receptor-mediated release from thalamic afferents. These findings have led to comparisons of the effects of classical hallucinogens with certain aspects of acute psychosis and to a focus on thalamocortical interactions as key to understanding both the action of these substances and the neuroanatomical sites involved in altered states of consciousness (ASC). In vivo brain imaging in humans using [(18)F]fluorodeoxyglucose has shown that hallucinogens increase prefrontal cortical metabolism, and correlations have been developed between activity in specific brain areas and psychological elements of the ASC produced by hallucinogens. The 5-HT(2A) receptor clearly plays an essential role in cognitive processing, including working memory, and ligands for this receptor may be extremely useful tools for future cognitive neuroscience research. In addition, it appears entirely possible that utility may still emerge for the use of hallucinogens in treating alcoholism, substance abuse, and certain psychiatric disorders. PMID- 14761704 TI - The genetic basis of high-altitude pulmonary oedema. AB - High-altitude pulmonary oedema (HAPE) is a potentially fatal condition affecting fit and previously well individuals at altitudes in excess of 3000 m. This article discusses the mechanisms of HAPE, considers the contribution of hypoxic pulmonary vasoconstriction and alterations in sodium transport to the pathological process. It discusses the various biochemical mediators such as nitric oxide (NO), endothelin-1 (ET-1), and the renin-angiotensin-aldosterone system (RAS) that may be involved and considers possible oxygen-sensing mechanisms involved in hypoxic adaptation such as hypoxia-inducible factor-1 (HIF 1). Those who have had HAPE once run an unpredictable but significant risk of recurrence; therefore, there may be a constitutional or genetic component in its aetiology. This paper considers the possible involvement of genes that may be involved in physiological adaptation to hypoxia (e.g., angiotensin-1 [AT(1)] converting enzyme [ACE], tyrosine hydroxylase, serotonin transporter [5-HTT], and endothelial NO synthase [eNOS] genes). As yet, no formal association has been identified between an identified genetic polymorphism and HAPE, but genetic variation provides a possible mechanism to explain interindividual variation in response to hypoxia and enhanced or reduced performance at altitude. PMID- 14761705 TI - [Current topics on marine medicinal products]. PMID- 14761706 TI - [Rotavirus nosocomial infection in pediatric units. A multicentric observation study]. AB - OBJECTIVE: Rotavirus nosocomial infection (RNI) is frequent in pediatric units. This study was designed to determine the incidence and the main risk factors of RNI in children aged 3 months-3 years and admitted for at least 48 hours days during the epidemic period. PATIENTS AND METHODS: A stool sample was obtained within the 24 hours of admission. An additional sample was collected from rotavirus-negative children either the day of discharge, or when they developed abnormal clinical signs. Parents were contacted by phone after discharge. Children initially rotavirus-negative and positive 2 days or more after admission were considered as certain nosocomial cases. In the absence of the second sample, possible nosocomial cases were considered if new symptoms (i.e.; fever and or digestive symptoms) occurred 2 days or more after the first negative sample. RESULTS: One hundred and seventeen children were included. The incidence was 11.1% for certain NRI, 16.8% for possible hospital-acquired cases and 19.4% for the whole cases. Possible risk factors were the low number of nurses during the weekend, the great number of medicine students in the unit, and no use of individual material. CONCLUSION: NRI have a high incidence, whose reality can only be approximated by taking into account the possible NRI occurring at home after hospital-discharge. PMID- 14761707 TI - [Rubella infection in children and teenagers in Tunisia]. AB - Rubella is a worlwide common infection; its importance in public health relates to the risk of malformation when primary infection occurs during pregnancy. This serosurvey was conducted to assess the kinetics of rubella infection in Tunisian children and teenagers and to determine the proportion of girls who remain seronegative at childbearing age. The studied population included 2481 individuals aged seven (N =1136), 13 (N =711) and 19 years (N =634), this sample was collected in 1996 and is representative of all geographical regions of the country. Our results indicate that 42% of tunisians are infected before seven years, 73% before 13 and 89% before 19 years of age. These rates are lower than those previously reported in the country. The proportion of seronegatives at 19 years of age was higher in costal regions than in the rest of the country: 14 vs 5% (p =0,0008). This difference should be due to the higher socio-economic level of the population living in costal regions. Our study indicates that primary infection with rubella virus in Tunisia is progressively shifting to older ages, which may increase the risk of congenital rubella syndrome. The introduction of rubella vaccination in the national program of vaccination may be considered, however only very high coverage levels will have a positive effect. Beside the reduction of the risk of congenital rubella syndrome, rubella vaccination will reduce the incidence of febrile rush cases and thus facilitate the surveillance activities conducted as part of the national program of measles elimination. PMID- 14761708 TI - [Nosocomial infections due to adenovirus in a paediatric unit]. AB - OBJECTIVE: We carried out a retrospective analysis of an outbreak of adenovirus (AdV) infections in a paediatric unit. The aim of the study was to analyse cases, determine the route of transmission and to evaluate the efficacy of the prevention measures. PATIENTS AND METHODS: The study was performed by recollection of AdV infection cases during a period of 1 year and the results were compared with the list of clinical cases recorded during the epidemic. The clinical files of children with a positive specimen were retrospectively analysed. During that period, five members of the medical staff showed clinical signs and symptoms of AdV infection. A throat swab was collected from a subset of the staff. RESULTS: Among nine patients with positive AdV detection, six were infected with an Adv type 2. Six were nosocomially-acquired, the other two were only probable nosocomial infections. The index case was a child presenting a febrile diarrhoea 48 h prior to being admitted to the hospital. Nosocomial transmission was associated with the prolonged shedding of the virus with faeces of the infected cases. The specimens collected from the staff remained negative. The outcome was favourable for all children. CONCLUSIONS: Prevention measures, implemented when the epidemic was characterised, allowed the control of the nosocomial outbreak. PMID- 14761709 TI - [Virulence factors of clinical Aeromonas caviae isolates]. AB - Aeromonas caviae, an ubiquitous aquatic organism, has long been considered to be of low pathogenicity, and its virulence mechanisms are still not clearly understood. Twenty-eight A. caviae isolates of clinical origin, most often monomicrobic, were identified in our university hospital over a four year period. Patients, mostly immunocompromised, were: eight diarrhoeal infants, 13 diarrhoeal adults, seven bacteraemic adults. Adults were frequently suffering from underlying intestinal malignancy, hepatobiliary disease, gastrectomy. Virulence factors were investigated. Adherence, studied by use of tissue culture HEp-2 cells, and staining of characteristic lateral flagella, were observed in diarrhoeal strains. Extracellular hemolytic activity was tested on rabbit erythrocytes suspensions at 25 and 37 degrees C. One blood culture isolate showed an important hemolytic activity at 25 degrees C, but none at 37 degrees C. Treatment with furin activated the aerolysin precursor and resulted in significant hemolysis at 37 degrees C, and fluid accumulation in rabbit ileal loops similar to that of A. hydrophila as control. The presence of the hemolysin gene was confirmed in this strain by PCR. In conclusion, A. caviae was shown to be a pathogen isolated from diarrhoea and bacteraemia in immunocompromised patients with malignancies and low gastric acidity as favouring factors. Virulence including the ability to adhere to cells and the production of lateral flagella was observed in diarrhoeal strains. The expression and the production of extracellular hemolytic activity and enterotoxicity at 37 degrees C depended on the activation of the pore forming toxin aerolysin precursor by furin. In vivo the protoxin is probably processed to its mature form by host proteases. PMID- 14761710 TI - [Accuracy of four agar diffusion methods and the Vitek 2 automated system for the detection of the methicillin resistance in coagulase negative staphylococci]. AB - AIM OF THE STUDY: Technical difficulties may occur in the detection of methicillin resistance in coagulase negative staphylococci. Phenotypic methods, such as disk diffusion testing, sometimes fail to detect methicillin resistance because of its poor expression. Four of these methods were compared with the VITEK 2 system. MATERIALS AND METHODS: The accuracy of the VITEK 2 system and the agar diffusion methods according to the current CASFM and NCCLS guidelines were assessed for methicillin susceptibility testing of 70 coagulase negative staphylococci isolates harboring the mecA gene for which the detection of the methicillin resistance was particularly difficult: the CASFM recommendations applied in 1998 failed to categorize them as methicillin-resistant. RESULTS: Among the different methods currently suggested by the CASFM, those using a heavy inoculum and a hypersaline medium give better results, but they remain however less efficient than the NCCLS recommendations testing (sensitivity: 71-86% vs. 94 94% respectively). The VITEK 2 was more efficient (sensitivity: 91%) than the disk diffusion methods recommended by the CASFM and slightly less efficient than the method recommended by the NCCLS. CONCLUSION: The VITEK 2 automated system, compared with the disk diffusion methods, showed adequate accuracy for detection of oxacillin resistance in coagulase negative staphylococci. PMID- 14761711 TI - [Recovery method of serotypable character in non serotypable pseudomonas aeruginosa strains]. AB - Serotyping is one of the most used techniques for typing Pseudomonas aeruginosa strains. During chronic infections, and especially in cystic fibrosis, the decrease of lipopolysaccharide production is responsible for difficulties in determining O antigens. The possibility of serotyping can be simply restored by using a primary culture broth containing amikacin (1/6 of the strain MIC for this antibiotic); this is due to the ability of this antibiotic to inhibit alginate production. This technique allowed us to determine the serotype of 108 non serotypable strains of P. aeruginosa isolated in 14 different hospitals. Among these isolates, serotype O:1 and O:13, had a high prevalence; the origin is a deficiency in D-glucose and L-rhamnose, required for the synthesis of lipopolysaccharide. In contrast, these sugars are not present in lipopolysaccharide of O:12, and these strains are always serotypable. The main protein is Alg C; this bifunctional enzyme is required in the exopolysaccharide and lipopolysaccharide production, according stress conditions in the bacterial cells' environment. Determination of the serotype, as Antibiogram, is essential for genotypic inquiries. PMID- 14761712 TI - [Role of anti-CD20 monoclonal antibody in association with immunomodulatory agents]. AB - Chimeric monoclonal anti-CD20 antibody (Rituximab) has been associated with immunomodulatory agents such as interferon alpha, interleukin-2, interleukin-12, G-CSF, GM-CSF and anti-CD22 humanized monoclonal antibody (Epratuzumab). Synergy with interferon is clearly demonstrated increasing complete response rate and response duration. Other associations are promising but must be tested in randomized prospective trials versus rituximab alone, probably in indolent lymphomas where chemotherapy could be avoided. PMID- 14761713 TI - Biological markers for the follow-up of athletes throughout the training season. AB - During the training season, a state of fatigue known as overtraining may occur, resulting from an excessive load of training, both in volume and intensity. Even now, difficult to predict the risk of overtraining, although this syndrome has been the subject of numerous studies. A lot of biological markers have been propounded. Taken alone, none of them have an absolute significance. This paper aims to review these markers, considering their biological interest, the ease with which they can be measured and the cost, from the simplest (body weight daily recording) to the most up to date markers (e.g. anti-oxidant status). They are grouped into three categories: non-invasive behavioural and biological markers, biochemical markers, and hormonal and immunological markers. PMID- 14761714 TI - [Transforming growth factor-betas: smad signaling and roles in physiopathology]. AB - Transforming growth factor-beta (TGF-beta) family members are multifunctional peptide growth factors that regulate cell growth, differentiation, extracellular matrix production and cell migration and embryonic development. Knock-out experiments for the three mammalian isoforms of TGF-beta in mice have demonstrated their importance in regulating inflammation and tissue repair. Also, TGF-beta has been implicated in the pathogenesis of human diseases, including tissue fibrosis and carcinogenesis. In the latter case, it may exert both tumor suppressor and pro-oncogenic activities depending on the stage of the tumor. Smads proteins constitute the core components of the intracellular signaling cascade initiated by TGF-beta receptors, as they carry signals from the cell surface directly to the nucleus; where they act as transcription factors. PMID- 14761715 TI - Verification of false-positive blood culture results generated by the BACTEC 9000 series by eubacterial 16S rDNA and panfungal 18S rDNA directed polymerase chain reaction (PCR). AB - A small but significant proportion of blood cultures processed by the BACTEC 9000 series systems is signaled positive, while subsequent Gram's stain and culture on solid media yield no pathogens. In this study, 15 "false-positive" vials (7 aerobes, 8 anaerobes) from 15 patients were investigated for the presence of bacteria and fungi by eubacterial 16S rDNA and panfungal 18S rDNA amplification, respectively. All samples turned out negative by both methods. Most patients (7) had neutropenia, which does not support the theory that high leukocyte counts enhance the generation of false-positive results. In conclusion, the results of this study indicate that false-negative results generated by the BACTEC 9000 series are inherent to the automated detection and not due to the growth of fastidious organisms. PMID- 14761716 TI - Antimicrobial susceptibility patterns, beta-lactamases, and biochemical identification of Yokenella regensburgei strains. AB - Yokenella regensburgei is an opportunistic human pathogen that phenotypically resembles Hafnia alvei. The susceptibility of 10 Y. regensburgei strains to 75 antimicrobial agents was examined, applying a microdilution procedure in cation adjusted Mueller-Hinton broth (CAMHB) and IsoSensitest broth (ISB). beta Lactamases were characterized phenotypically with beta-lactamase activity and induction assays. Genotypically, PCR experiments applying degenerated primer pairs for the detection of AmpC beta-lactamase genes were performed. Examining the phenotypic properties of Yokenella and 76 H. alvei strains with commercial identification systems and conventional tests, a database for an accurate biochemical separation of Y. regensburgei from H. alvei was established. In CAMHB, all tested yokenellae were resistant or at least of intermediate susceptibility to penicillin G, oxacillin, amoxicillin, amoxicillin-clavulanate, cefaclor, cefazoline, loracarbef, cefoxitin, all tested macrolides, lincosamides, streptogramins, ketolides, fusidic acid, glycopeptides, linezolid, and rifampicin. All Yokenella strains were sensitive to several beta-lactams, all tested aminoglycosides, chloramphenicol, folate-pathway inhibitors, fosfomycin, nitrofurantion, quinolones, and tetracyclines. In ISB, the minimum inhibitory concentration (MIC) values of several beta-lactams were one to four MIC doubling dilution steps lower than those found in CAMHB (depending on the beta-lactam). All yokenellae yielded specific amplification products for ampC, and all of these strains expressed beta-lactamases that were strongly inducible. Hydroxyproline amidase, maltosidase, tri-peptidase, proline deaminase, catalase reaction, Voges Proskauer test, and fermentation of glycerol, melibiose and myo-inositol were suitable parameters to separate Y. regensburgei from H. alvei. PMID- 14761717 TI - Clinical experience with Staphylococcus lugdunensis bacteremia: a retrospective analysis. AB - We report a retrospective review of all cases of Staphylococcus lugdunensis bacteremia identified during a period of more than 10 years (January 1990 to May 2002) in a large medical center. Twenty-one cases occurred over that period; 20 cases were available for review. In six instances, the organism appeared to be clinically significant and was most commonly related to a vascular line infection. No deaths, secondary suppurative complications, prolonged bacteremia, or prolonged fever appeared to be caused by S. lugdunensis in our experience. Twenty-five percent of our isolates were resistant to multiple antibiotics, including oxacillin. PMID- 14761718 TI - Low-density addressable array for the detection and typing of the human papillomavirus. AB - We have developed a low-density DNA array for the detection and typing of human papillomavirus (HPV) DNA. The gene chemistry strategy involves using a combination of the polymerase chain reaction (PCR) with the consensus oligonucleotide primers MY09/MY11 followed by a ligase detection reaction (LDR). Fluorochrome-labeled HPV-specific primers are joined to a common primer modified with a unique anchoring sequence called a zip code on its 3' end. The result is a series of 60-70 base pair and single-stranded ligation products that are then hybridized to their respective zip code complements affixed to glass slide based arrays. Nine separate zip codes were assigned, one for each HPV type (6,11,16,18, 31, 33, 35, and 53) and one for a beta-globin internal control marker. Two additional zip-codes were reserved for a pair of consensus HPV LDR products: the cLDR1 and cLDR2 primers hybridize to a conserved sequence within the HPV L1 open reading frame internal to the MY09/MY11 fragment. These consensus primers were shown to detect over 40 different HPV types. The purpose of this study was to evaluate the analytic performance of this low-density microarray based assay for HPV, as well as to introduce our simplified read-out instrumentation, shown here to be a low cost and highly efficient way to detect and genotype HPV for clinical testing. PMID- 14761719 TI - Distribution and antifungal susceptibility of Candida species causing candidemia from 1996 to 1999. AB - Susceptibilities to amphotericin B and fluconazole of 383 Candida species isolated from blood were determined. Candida albicans was the most common species (55.6%), followed by Candida parapsilosis (17.5%), Candida tropicalis (16.5%), Candida glabrata (5.2%), Candida guilliermondii (2.3%), and others (2.9%). All but three isolates, Candida ciferrii, C. tropicalis, and C. glabrata, one each, were susceptible to amphotericin B. A total of 367 (95.8%) and 15 (4.2%) isolates were susceptible and susceptible-dose dependent to fluconazole, respectively. Only one isolate, a C. glabrata, was resistant to fluconazole. Few patients (13%) having prior fluconazole treatments may explain the low rate of resistance to fluconazole in this study. PMID- 14761720 TI - Evaluation of four DNA extraction methods for the detection of Mycobacterium avium subsp. paratuberculosis by polymerase chain reaction. AB - Polymerase chain reaction (PCR) has been widely used due to its high specificity, sensitivity, and rapid turn-around time. However, inhibitory factors may be co extracted with the target nucleic acid that will hinder the performance of PCR. In this study, DNA extraction methods for Mycobacterium avium subsp. paratuberculosis were evaluated including rapid lysis, organic extraction, silica based and magnetic particle-based (MagaZorb) technologies on bacterial cells, and spiked bovine feces. Efficiency of the extraction was determined by PCR end point titration with primers targeting the insertion sequence, IS900. Results of the end point titrations are identical for bacterial cells and spiked feces. Inhibition was observed in PCR with DNA isolated from spiked feces, and a 1/100 dilution was able to alleviate this problem with DNA extracted by MagaZorb. A 1/1000 dilution was required for the other three methods. MagaZorb proved to be more efficient at removing inhibitory factors and required the least labor and completion time. Further evaluation is required for its utilization in other clinical specimens. PMID- 14761721 TI - Detection of rifampin-resistant Mycobacterium tuberculosis strains by using a specialized oligonucleotide microarray. AB - DNA microarray represents one of the major advances in diagnostic sequencing of polymerase chain reaction (PCR) products. Until now, arrays have been relatively expensive, complex to perform, and difficult to interpret, limiting their wide application in the clinical laboratory. A moderate-density oligonucleotide microarray that can rapidly identify Mycobacterium tuberculosis rifampin resistant strains was developed. The method is based on the detection of point mutations and other rearrangements in the rpoB gene region determining rifampin resistance. Rifampin resistance was determined by hybridizing fluorescently labeled, amplified genetic material generated from bacterial colonies to the array. Fifty-three rifampin-resistant M. tuberculosis and 15 rifampin-susceptible M. tuberculosis were tested and results were concordant with those based on culture drug susceptibility testing and sequencing. Rifampin-resistant clinical isolates were detected in as little as 1.5 hours after PCR amplification with visual results. It is demonstrated that oligonucleotide microarray is an efficient, specialized technique to implement and can be used as a rapid method for detecting rifampin resistance to complement standard culture-based method. PMID- 14761722 TI - Disk diffusion quality control guidelines for NVP-PDF 713: a novel peptide deformylase inhibitor. AB - NVP-PDF713 is a peptide deformylase inhibitor that has emerged as a candidate for treating Gram-positive infections and selected Gram-negative species that commonly cause community-acquired respiratory tract infections. This report summarizes the results of a multi-center (seven participants) disk diffusion quality control (QC) investigation for NVP PDF-713 using guidelines of the National Committee for Clinical Laboratory Standards and the standardized disk diffusion method. A total of 420 NVP-PDF 713 zone diameter values were generated for each QC organism. The proposed zone diameter ranges contained 97.6-99.8% of the reported participant results and were: Staphylococcus aureus ATCC 25923 (25 35 mm), Streptococcus pneumoniae ATCC 49619 (30-37 mm), and Haemophilus influenzae ATCC 49247 (24-32 mm). These QC criteria for the disk diffusion method should be applied during the NVP-PDF 713 clinical trials to maximize test accuracy. PMID- 14761723 TI - The significance of combining World Health Organization and Center for Disease Control criteria to resolve indeterminate human immunodeficiency virus type-1 Western blot results. AB - A total of 466 repeatedly reactive HIV-1 blood samples evidenced by enzyme immunoassay were analyzed by the Western blot method and interpreted according to WHO and CDC criteria. Discordant and indeterminate samples were further analyzed by PCR. When the Western blot result was classified as indeterminate, according to both WHO and CDC criteria, the PCR test was always negative. These findings suggest that samples with double-indeterminate status should be reported as negative. PMID- 14761724 TI - Recurrent Listeria monocytogenes aortic graft infection: confirmation of relapse by molecular subtyping. AB - Based on molecular typing methods, we identified a rare case of a recurrent L. monocytogenes infection resulting from an infected aortic prosthesis as detected by 18-F-Fluoro-d-deoxyglucose positron emission tomography (FDG PET). Our case highlights the usefulness of molecular typing and nuclear imaging methods for understanding L. monocytogenes pathogenesis and epidemiology. PMID- 14761725 TI - In vitro interactions of licensed and novel antifungal drugs against Fusarium spp. AB - We have studied the in vitro interactions of amphotericin B (AMB) with terbinafine (TBF), itraconazole, voriconazole (VCZ), albaconazole, and ravuconazole (RVZ), as well as TBF combined with the same azoles, against 11 isolates of Fusarium spp. using the fractional inhibitory concentration index. The highest percentage of synergistic interactions was observed for the combinations AMB-RVZ, TBF-VCZ, and TBF-RCZ. PMID- 14761727 TI - [French pediatric oncology: past and future]. PMID- 14761726 TI - In vitro activity of BAL9141 against clinical isolates of gram-negative bacteria. AB - The minimum inhibitory concentration and minimum bactericidal concentration of BAL9141 against 150 clinical isolates of gram-negative bacteria were determined and compared to those of cefepime and ceftriaxone. BAL9141 exhibited comparable activity to cefepime and ceftriaxone against Haemophilus influenzae, Klebsiella pneumoniae, and extended-spectrum beta-lactamase nonproducing Enterobacter cloacae, and comparable activity to cefepime against Pseudomonas aeruginosa. PMID- 14761728 TI - [A comparative study of nocturnal sleep duration and timetable of children between five- and ten-years-old according to their age and socio-economic environment]. AB - AIM: This study was conducted on a sample of 644 pupils between the ages of five and ten years at school in the Orleans-Tours education authority with the aim of studying the nocturnal sleep duration and timetable of young children according to age and socio-economic environment. METHOD: In order to find out about children's sleeping habits (duration, time of going to bed and getting up, and weekly variations) a standard grid was used to carry out a cross-sectional survey in the children's families. This was filled in each day from Monday to Sunday during the same school week for all the children. RESULTS: Sleep duration decreased with age from maternelle to CM2 (nursery to last year of primary education). Data relating to sleep duration from CE2 (third year of primary school - 8-year-old -) showed differences according to whether the school belonged to an Educational Priority Zone (EPZ) or not. It was noticed that between five- and ten-year-old children from EPZ lost 62 minutes of sleep, whereas those not from EPZ only lost 29 minutes. These results would suggest that in addition to developmental factors, environmental factors also play a role in sleep duration. The differences observed were due to later bed times for children from EPZ. Weekly variations in sleep were generally very similar for all the children. At the weekend all the children tended to go to bed later, however this was more noticeable Saturday night for children not living in EPZ. Children slept the longest on Tuesday night due to the fact they got up later Wednesday morning (Wednesday is a day off in the majority of French schools). However, children from the age of nine (CM1 - forth year of primary education -) in EPZ did not benefit from this recuperation time, as they went to bed later but still got up early the next morning. CONCLUSION: This study showed that in addition to the physiological and developmental factors that influence children's sleep, the socio-economic context also plays a role. These results as a whole highlight the importance that practitioners and families should pay to maintaining a regularity in the child's routine and in the amount of sleep necessary at each age. PMID- 14761729 TI - [Sleep problems and bedtime routines in infants in a cross cultural perspective]. AB - Intercultural studies induced many controversies in the characteristics associated with sleep problems. OBJECTIVES: The aim of this study was three fold: (1) to compare sleep problem frequency in two culturally different populations; (2) to analyze if some particular bedtime routines ("co-sleeping", "distal" or "proximal" parent-infant interaction) are associated to trouble; (3) to show influence of the cultural context to trouble. POPULATION: This study examines bedtime routines in French infants aged 12 to 24 months in maternal and infant's protect center (PMI) in Lyon and la Reunion island, for a sleep - cross cultural approach. Three hundred infants were enrolled in the survey. RESULTS: We showed prominent sleep problems in la Reunion's group (35%) in comparison with the group in Lyon (17%) (P <0.005). Regression analysis allowed to draw relations between several factors and the sleep problems. Hence, proximal bedtime routines and interactions (CR =0.530, P =0.05), "co-sleeping" (CR =0.989, P =0.005) and parental presence at bedtime until infant falls asleep (CR =0.595, P =0.04) are significantly associated with sleep problems. These attitudes are more frequent in la Reunion's group. CONCLUSION: Our study confirms literature data and asks question about cultural meaning for these inadequate behaviors in relation to mother-infant separation distress. Acculturation notion is called forth in la Reunion's group. PMID- 14761730 TI - [Vincristine treatment for function- and life-threatening infantile hemangioma]. AB - AIM: To evaluate the efficacy of vincristine treatment for function- and life threatening hemangiomas. PATIENTS AND METHOD: Nine infants, eight girls and one boy, received vincristine treatment (VCR) for endangering hemangiomas. In six cases, the hemangiomas involved head and neck in a segmental unilateral or bilateral distribution (3/6 also had laryngeal and 2/6 tracheal location causing respiratory distress, 5/6 had eyelid and orbital involvement); one infant had disseminated neonatal hemangiomatosis (skin, liver, kidney); two infants had liver hemangiomas with cardiac failure. VCR was prescribed after failure of high dosage corticosteroid treatment in six, and of both corticosteroids and interferon alpha 2b (5 months) in one; two infants received VCR as first line treatment. RESULTS: A dosage of 1 mg/m(2) IV injection was delivered, with weekly injections first, and then tapering, increasing the interval between injections, depending on the clinical response. The nine infants received from 5 to 25 injections (average: 16), for a length of treatment of 1.5-8 months (average: 5.5 months). In seven patients a clear clinical response was observed at the end of the first month of treatment, while a slow protracted response was noted in two. Transient mild side effects were present in four patients. DISCUSSION: Corticosteroid treatment, although a worldwide recognized treatment of problematic hemangiomas, cannot always control the growth of alarming hemangiomas. Interferon alpha 2a and 2b have proven a 90% effectiveness: treatment for cortico-resistant, function- and life-threatening, hemangiomas. PMID- 14761731 TI - [Acute paediatric care and unplanned consultations - a survey in health care facilities in the "Grand Lyon" area]. AB - With the increasing number of emergency paediatric consultations, two surveys "on a given day" were performed in the "Communaute Urbaine de Lyon", with the aim of analyzing the causes, circumstances, and relevance of these unplanned consultations, as well as a better understanding of the itinerary of these patients. METHODS: Unplanned consultations concerned children less than 18 years old seen in consultation on Saturday April 21, and Thursday December 13, 2001, in all medical facilities capable of delivering emergency paediatric care. Data collection was performed by filling out a questionnaire given to the family and the physicians (general practitioner who were chosen randomly, or paediatricians volunteering for the study), working in general medicine sectors, at the outpatient emergency consultation of the "Groupement des Pediatres du Lyonnais", and in all the emergency departments to which children could be addressed (public hospitals and private clinics). "SOS Medecins" did not participate in this study but communicated its activity a posteriori. Six hundred and eighty three consultations on the Saturday and 1183 on the Thursday were analyzed. RESULTS: An estimation of the total number of consultations was performed taking into account the proportion of practitioners participating in the survey, with a total number of 1813 consultations on the Saturday and 4576 on the Thursday. Consultations in the private setting (by practitioners or organized emergency centers) accounted for 82% on the Saturday (general practitioners 70%, paediatricians 12%) and 93% on the Thursday (general practitioners 75%, paediatricians 18%), public hospitals 13% and 4% and private clinics 5% and 2%, respectively. Parents considered the problem to be serious in 10-40% of the cases, depending on the setting. The reason justifying consultation was generally medical (pain or discomfort, fear of complication, less frequently feeling of imminent danger). Non-medical reasons (proximity of the week-end, personal reasons, absence or unavailability of usual practitioner) concerned a fourth of the Thursday consultations and up to half of the Saturday consultations. An urgent consultation was estimated not be justified in 13% of the cases according to the physicians. The main reason for going to a hospital was that "everything would be available on site". On Saturday the absence of the usual practitioner was the most cited reason. The global itinerary was deemed satisfactory by the physicians in 82% of the cases. CONCLUSION: Unplanned consultations are found in three situations: routine and accepted activity of medical office (consultation without an appointment), urgent medical problem (or estimated to be urgent), and the result of a dysfunction between the medical possibilities of the health care system and its use by the parents. This last point opens two possibilities of action which are the information and education of families and the networking of physicians involved in emergency consultations with the aim of reducing hospital consultations by 15-20%. PMID- 14761732 TI - [Cerebro-costo-mandibular syndrome. A case-report]. AB - The cerebro-costo-mandibular syndrome is a rare and serious congenital disorder characterized by the association of posterior rib malformations, micrognatia and mental deficiency. Death due to respiratory failure is usual during the first days or months of life. The various clinical and radiological aspects of this disease are described based on a recent observation. PMID- 14761733 TI - [Mitochondrial cytopathy: an unusual infantile cause of total villous atrophy]. AB - Villous atrophy in an infant immediately suggests food intolerance. We report a case with an unusual cause. This female infant was first examined at 5 months for anorexia and failure to thrive. Intestinal biopsy showed total villous atrophy. A diet excluding gluten and cow milk proteins failed to improve her condition and the infant was hospitalized at 7 months for further investigations. The infant was hypotonic with a head lag. No other clinical sign was noted. Serum transaminases were increased 5- to 10-fold and CSF proteins concentration was increased. Metabolic investigations revealed hyperlactacidaemia and an increased lactate/pyruvate ratio during fasting and feeding, suggesting a mitochondrial cytopathy. Respiratory chain enzymatic activity measurements confirmed the diagnosis and showed severely decreased activities of complexes I, III and IV in both the liver and muscle. Molecular analysis demonstrated depletion of mitochondrial DNA in the liver (75%) and in muscle (97%). The infant was discharged under continuous enteral nutrition. Improvement was of short duration and the infant died at 1 year of age of massive hepatic failure. This is the first report of a mitochondrial DNA depletion with total villous atrophy and malabsorption as early clinical onset. A mitochondrial cytopathy should be considered in such conditions when food exclusion diets fail. PMID- 14761734 TI - [Streptococcus pyogenes endocarditis following varicella: a case report]. AB - Although varicella is most often a benign and self-limited disease of childhood, it can be associated with a variety of serious and potential lethal complications. Especially, the incidence of severe infectious complications caused by group A streptococci has been increasing over the last years. We report the case of a previously healthy young boy with an aortic bicuspidy who developed a varicella complicated by endocarditis due to group A streptococcus, and a haemophagocytic syndrome. A favorable outcome was obtained after an early valvular replacement and 6 weeks of intravenous antibiotics. PMID- 14761735 TI - [Mixed connective tissue disease revealed by chronic lymphocytic meningitis in an infant]. AB - Mixed connective tissue disease (MTCD) is a systemic inflammatory disorder individualised by Sharp et al. in 1972. This entity is rare in children. CASE REPORT: We report an exceptional case of MTCD revealed by lymphocytic meningitis in a two-month-and-a-half-old infant. The disease was diagnosed at the age of nine months when clinical symptomatology was completed by common signs of the illness (Raynaud's phenomenon, swollen hands), systemic lupus erythematosus-like symptoms (lymphadenopathy, squamous erythema of the limbs, hepato-splenomegaly, pleuritis and ascites) and polymyositis-like findings (muscle weakness with increased serum levels of myogenic enzymes). Laboratory investigations showed an important inflammatory syndrome and the presence of speckled anti-nuclear and anti-U(1)RNP antibodies. Specific antibodies of the other connective tissue diseases were also positive (anti-DNA, anti-Sm, anti-SSA and SSB, anti-Scl 70 and JO1) pleading for the mixed feature of the illness. The follow-up after corticosteroid treatment was marked by clinical and biological improvement. But after five months, the patient died following a severe infectious complication. CONCLUSION: Chronic lymphocytic meningitis can be part of Sharp's syndrome even in infants. However, the diagnosis relies on the evidence of characteristic clinical and biological abnormalities of MTCD. PMID- 14761736 TI - [Radiological quiz of the month: a case of vomiting in a patient treated for kyphoscoliosis by orthopaedic corset]. PMID- 14761737 TI - [Acute pancreatitis and Mycoplasma pneumoniae infection]. PMID- 14761738 TI - [Obesity of child in two Dakar hospitals: epidemiological aspects and problems of management]. PMID- 14761739 TI - [Tumoral pathology in foreign children in Abidjan]. PMID- 14761740 TI - [Post-traumatic lower limb pain in children]. AB - Injuries to the lower extremity are frequent. They present as pain, reduction of joint function, alteration in gait. Most of the orthopaedic emergencies for teenagers are related to trauma. For younger children, a history of recent fall is prevalent but the non traumatic orthopaedic conditions are especially important to investigate when the reported injury is minor or when onset of symptoms is delayed. PMID- 14761741 TI - [The pediatrician's emergency box]. AB - Every private pediatrician must be available to respond at pediatrics emergencies over thinking action in acute situations, which can quickly worsen. With his knowledge and a vigorous experience on field, he must possess minimum material and drugs, put in an emergency box to talking face up to distress's situations really or feeling like that by parents or in close circle of child. Private pediatrician is a link of a care chain and/or assistance, to be waiting that wounded arrive to hospital emergency unit or in a specialized service, when he will be taken in charge by SMUR team. PMID- 14761742 TI - [Cardiac syncope in children]. AB - Syncope is a frequent problem in childhood; generally, it is an isolated event and the common causes are benign. However, in some circumstances, syncope can herald a potentially lethal problem, especially when occurring during exercise. Routine evaluation includes history, physical examination and a 12-lead standard ECG should be performed in all cases. Worrying features which should be an indication for further investigation include syncope during exercise, collapse in a swimming pool, history of familial sudden death, and abnormalities on clinical exam or ECG. Structural cardiac abnormalities that may cause syncope and sudden death include aortic stenosis, hypertrophic cardiomyopathy and coronary malformations. All children with unrepaired or repaired congenital heart disease who experienced a syncope should be referred to a specialist. Primary arrhythmias that are easily diagnosed on ECG are the long QT syndrome, complete atrio ventricular block and Wolff-Parkinson-White syndrome; ST elevation in V1-V3 may reveal a Brugada syndrome. Another arrhythmia which is known to be potentially fatal if undiagnosed is the catecholaminergic ventricular tachycardia; the baseline ECG is normal but the arrhythmia is easily reproduced during exercise testing. Finally, vasovagal syncope is the most likely cause of syncope in the young and it usually easily recognized. PMID- 14761743 TI - [Does selective digestive decontamination prevent nosocomial infections?]. AB - Paediatric intensive care and haematological units are ideal sites for the development of nosocomial infections. These infections remain a significant source of mortality and morbidity and increase length of stay and costs. Selective digestive decontamination (SDD) includes topical antibiotics during the entire intensive care unit (ICU) stay, parenteral antibiotic administered for three to five days, hand hygiene and surveillance cultures of throat and rectum. Its use is based on the observation that resistant bacteria are often imported by the patients themselves, and the fact that transmission via the hands of carers could be responsible only for infections occurring after one week. In adult patients, seven meta-analyses have demonstrated that SDD reduces the odds ratio for lower airway infections, and sometimes mortality (particularly in surgical and trauma patients). The main criticism against SDD is the possible emergence of antibiotic resistant bacteria, which is a growing problem in Europe and United States of America. Only four studies on SDD in children have been reported in the literature: due to methodological weaknesses and small size of samples, definitive conclusion cannot be drawn. However, one study in a 20 bed paediatric intensive care unit has demonstrated that SDD prevent both infections and the emergence of resistant bacteria. Furthermore, it has been demonstrated that more than 50% of children carrying resistant bacteria are detected within 24 hours of admission, suggesting that they import the resistant strains onto the intensive care unit. Factors that predict facility, administration of i.v. antibiotics within the past 12 months, previous intensive care unit admission and hospitalization of a household contact within the past 12 months. As suggested by several authors, the term selective should mean selection of appropriate patient groups (those at high risk of nosocomial infection, e.g. patients mechanically ventilated for at least 48 hours) and units (excluding those where multiresistance is endemic). Obviously, surveillance of patient and unit bacterial ecology and improvement of antibiotic policy must be reinforced. PMID- 14761744 TI - [Tuberculosis]. PMID- 14761750 TI - Recovery of aluminium, nickel-copper alloys and salts from spent fluorescent lamps. AB - This study explores a combined pyro-hydrometallurgical method to recover pure aluminium, nickel-copper alloy(s), and some valuable salts from spent fluorescent lamps (SFLs). It also examines the safe recycling of clean glass tubes for the fluorescent lamp industry. Spent lamps were decapped under water containing 35% acetone to achieve safe capture of mercury vapour. Cleaned glass tubes, if broken, were cut using a rotating diamond disc to a standard shorter length. Aluminium and copper-nickel alloys in the separated metallic parts were recovered using suitable flux to decrease metal losses going to slag. Operation variables affecting the quality of the products and the extent of recovery with the suggested method were investigated. Results revealed that total loss in the glass tube recycling operation was 2% of the SFLs. Pure aluminium meeting standard specification DIN 1712 was recovered by melting at 800 degrees C under sodium chloride/carbon flux for 20 min. Standard nickel-copper alloys with less than 0.1% tin were prepared by melting at 1250 degrees C using a sodium borate/carbon flux. De-tinning of the molten nickel-copper alloy was carried out using oxygen gas. Tin in the slag as oxide was recovered by reduction using carbon or hydrogen gas at 650-700 degrees C. Different valuable chloride salts were also obtained in good quality. Further research is recommended on the thermodynamics of nickel copper recovery, yttrium and europium recovery, and process economics. PMID- 14761751 TI - Field trial of a new aeration system for enhancing biodegradation in a biopile. AB - The influence of a new aeration system on the biopile performance was investigated. The purpose was to increase biodegradation efficiency by optimising airflow through the pile. During a 1-month field trial, the performance of a new system using two perforated vertical pipes with wind-driven turbines was compared with that of a standard pile configuration with two horizontal perforated pipes. Both piles were composed of a similar mix of diesel-contaminated soils, woodchips, compost and NPK fertiliser. Hydrocarbons were recovered using solvent extraction, and determined both gravimetrically and by gas chromatography. Total heterotrophs, pH and moisture content were also assessed. Air pressure measurements were made to compare the efficiency of suction in the pipes. Results at the end of the experiment showed that there was no significant difference between the two piles in the total amount of hydrocarbon biodegradation. The normalised degradation rate was, however, considerably higher in the new system than in the standard one, suggesting that the vertical venting method may have improved the efficiency of the biological reactions in the pile. The pressure measurements showed a significant improvement in the suction produced by the new aeration system. However, many factors other than the airflow (oxygen supply) may influence and limit the biodegradation rates, including moisture content, age of contaminants and the climatic conditions. Additional experiments and modelling need to be carried out to explore further the new aeration method and to develop criteria and guidelines for engineering design of optimal aeration schemes in order to achieve maximum biodegradation in biopiles. PMID- 14761752 TI - Effects of basaltic mineral fines on composting. AB - A by-product of the construction aggregate industry is fines or dust that contain trace elements such as zinc and copper and significant amounts of iron, aluminum, silica and potassium. Beneficial uses for these materials have been proposed such as replenishing depleted soils and amendment in mixtures of organic byproducts prior to composting. To evaluate the beneficial uses in composting, outdoor bin studies were conducted using a beef cattle manure, straw and wood chip mixture amended with and without basaltic mineral fines. Temperature differences in composting mixtures of equal volumes, equal moisture and relatively equal material content are considered an indication of differing biological activities [Haug, Compost Engineering Principles and Practice. Ann Arbor Science, Ann Arbor, MI. (1980)]. Temperatures were lower in the mineral fine-treated manure mixture initially. After turning the piles at six weeks, temperatures tended to be higher in the mineral fine amended mixture. Overall, temperatures were not significantly different suggesting that mineral fines amendment does not significantly increase temperature and activity in composting mixtures. PMID- 14761753 TI - Alternatives for treatment and disposal cost reduction of regulated medical wastes. AB - Many hospitals or health care facilities have faced financial difficulties and thus they have attempted to find cost-effective treatment and disposal methods of their regulated medical wastes (RMWs). This study investigated generation volume and sources, composition, and treatment and disposal methods for RMWs obtained from three out of the five typical city hospitals in Massachusetts for which we could obtain relevant data on medical waste. Also, this study compared the generation patterns and amounts of RMWs between the hospital and the medical school. The yearly operational treatment and disposal costs of RMWs based on different treatment and disposal methods were analyzed for one hospital. The most cost-effective option of four different treatment and disposal options studied was to combine on-site incineration and microwave technologies. Finally, this study identified measures for the effective waste characterization methods for the reduction of treatment and disposal costs of RMWs. By careful exclusion of non-RMW from RMW waste streams, hospitals can reduce the RMW volume that requires special treatment and reduce disposal costs. PMID- 14761754 TI - Use of rubber and bentonite added fly ash as a liner material. AB - In many countries regulations require all hazardous waste disposal facilities to be lined with suitable impermeable barriers to protect against contamination. In this study, a series of laboratory tests on rubber and bentonite added fly ash were conducted. The aim of the tests was to evaluate the feasibility of utilizing fly ash, rubber and bentonite as a low hydraulic conductivity liner material. Type C fly ash was obtained from Soma thermal power plant in Turkey; rubber in pulverized form was waste from the retreading industry. To investigate the properties of rubber and bentonite added fly ash, hydraulic conductivity, leachate analysis, unconfined compression, split tensile strength, one dimensional consolidation, swell and freeze/thaw cycle tests were performed. The overall evaluation of results have revealed that rubber and bentonite added fly ash showed good promise and a candidate for construction of a liner. PMID- 14761755 TI - Preliminary assessment of three new European leaching tests. AB - Leaching tests are used for characterization purposes and are indispensable for characterization of waste- and building materials. In several countries the use of leaching tests is settled by legislation. Since Europe is becoming a more united community, legislation becomes more and more harmonized as a logical next step in a more European approach towards environmental issues. In this study a comparison is made between three leaching tests, which are being developed in CEN technical committee 292. The percolation test is basically a column filled with granulated material, which is percolated with simulated rainwater for 3 weeks. The amount of water corresponds to 10 times the amount of solid; the eluate is sampled in several fractions. The second test is the pH-stat test in which pulverized material is leached for 24 h at eight different pH-values with the same liquid/solid ratio of 10. The third test reported is the shake test, in which the maximum leachable amount is being investigated at the materials own pH value. For this purpose pulverized material was used and the total amount, of water had again the same liquid/solid ratio of 10. A comparison was made in exchangeability of produced data. The results of the leaching experiments were comparable within limits of +20 to -43%. Other matters such as running time, information generated about leaching behavior and cost are also discussed. The pH stat test gives the most information, but is expensive. The percolation test generates detailed information, whereas the shake test is a relatively inexpensive alternative for the percolation test. PMID- 14761756 TI - Pyrolysis of virgin and waste polypropylene and its mixtures with waste polyethylene and polystyrene. AB - A comparison of waste and virgin polypropylene (PP) plastics under slow pyrolysis conditions is presented. Moreover, mixtures of waste PP with wastes of polyethylene (PE) and polystyrene (PS) were pyrolyzed under the same operating conditions. Not only the impact of waste on degradation products but also impacts of the variations in the mixing ratio were investigated. The thermogravimetric weight loss curves and their derivatives of virgin and waste PP showed differences due to the impurities which are dirt and food residues. The liquid yield distribution concerning the aliphatic, mono-aromatic and poly-aromatic compounds varies as the ratio of PP waste increases in the waste plastic mixtures. In addition to this, the alkene/alkane ratio of gas products shows variations depending on the mixing ratio of wastes. PMID- 14761757 TI - An investigation of halogens in Izmit hazardous and clinical waste incinerator. AB - In the combustion facilities, halogens (Cl, F, Br, I) should be considered with regard to the control of the compounds such as polychlorinated dibenzodioxins (PCDD), polychlorinated dibenzofurans (PCDF), halogenated polyaromatic hydrocarbons (PAH), polychlorinated biphenyls (PCB) and volatile heavy metals formed as a result of incomplete combustion and caused adverse environmental effects. In this study halogens were observed in Izmit Hazardous and Clinical Waste Incinerator (IZAYDAS). Halogen contents of the combustion menu, flue gas, fly ash, bottom ash and filter cake were measured and their distributions in these exit streams were determined. Results showed that the major part of the halogens was partitioned to solid residues, i.e., bottom ash and filter cake which represents the removal by wet scrubbers. Fly ash and flue gas fractions of halogens were much lower due to the reduced formation of volatile compounds. PMID- 14761758 TI - Fate of heavy metals and radioactive metals in gasification of sewage sludge. AB - The fates of radioactive cadmium, strontium, cesium, cobalt, arsenic, mercury, zinc, and copper spiked into sewage sludge were determined when the sludge was gasified by a process that maximizes production of char from the sludge (ChemChar process). For the most part the metals were retained in the char product in the gasifier. Small, but measurable quantities of arsenic were mobilized by gasification and slightly more than 1% of the arsenic was detected in the effluent gas. Mercury was largely mobilized from the solids in the gasifier, but most of the mercury was retained in a filter composed of char prepared from the sludge. The small amounts of mercury leaving the gasification system were found to be associated with an aerosol product generated during gasification. The metals retained in the char product of gasification were only partially leachable with 50% concentrated nitric acid. PMID- 14761759 TI - The hydration characteristics and utilization of slag obtained by the vitrification of MSWI fly ash. AB - This study investigated the effects of slag composition on the hydration characteristics of slag blended cement (SBC) pastes. Synthetic slag samples were prepared by melting CaO-modified and Al(2)O(3)-modified municipal solid waste incinerator (MSWI) fly ash. MSWI fly ash was mixed with 5% CaO and 5% Al(2)O(3) (by weight), respectively, resulting in two fly ash mixtures. These mixtures were then melted at 1400 degrees C for 30 min to produce two types of slag with different contents, designated at C-slag and A-slag. Both the C-slag and A-slag samples exhibited a pozzolanic activity index higher than the unmodified slag sample. The results show that the synthetic slags all met the Taiwan EPA's current regulatory thresholds. These synthetic slags were then blended with ordinary Portland cement (OPC) at various weight ratios ranging from 10 to 40%. The 28-day strength of the C1 paste was higher than that developed by the OPC paste, suggesting that the C-slag contributed to the earlier strength of the SBC pastes. At curing times beyond 28 days, the strength of the A1 paste samples approached that of the OPC paste samples. It can be seen from this that increasing the amount of calcium and aluminum oxide increases the early strength of SBC. The C-slag blended cement paste samples showed an increase in the number of fine pores with the curing time, showing that the C-slag enhanced the pozzolanic reactions, filling the pores. Also, the incorporation of a 10% addition of C-slag also tended to enhance the degree of hydration of the SBC pastes during the early ages (3-28 days). However, at later ages, no significant difference in degree of hydration between the OPC pastes and the SBC pastes was observed with the 10% C-slag addition. However, the incorporation of A-slag did decreased the degree of hydration. A slag blend ratio of 40% significantly decreased the hydration degree. PMID- 14761760 TI - Cement-clay pastes for stabilization/solidification of 2-chloroaniline. AB - Immobilization of a model liquid organic pollutant, i.e. the 2-chloroaniline (2 CA), into a cement matrix using organoclays as pre-sorbent agents was investigated. Five cement-clay pastes were prepared with different nominal water to-cement ratios (w/c=0.40, 0.25 and 0.15 wt/wt) and various amounts of waste (waste-to-cement o/c=0.20, 0.60 and 1.00 wt/wt); for comparison, a neat cement paste was also prepared. Dynamic leach tests were performed on solidified monoliths in order to assess the successful immobilization of the 2-CA. In monoliths at constant w/c ratio (0.40) the total amount of pollutant released increases with its initial content, and ranges from 15 to 35% with respect to it. By lowering w/c from 0.40 to 0.15 at constant o/c, the performances improved (<25% released). The microstructure of the hardened cement-clay pastes was characterized by quantitative X-ray diffraction (QXRD) and electronic microscopy (SEM-EDS) techniques; hydration degree was estimated by means of thermogravimetric analysis (TGA) in addition to QXRD. No evidence of any chemical reaction between 2-CA and cement phases was found. Moreover, it was shown that the most important factors affecting the cement hydration process were the total water content, i.e. the one taking also into account the water contained in the wet polluted clay, and the amount of 2-CA not firmly sorbed by the organoclay, and then freely dispersed in the paste. PMID- 14761761 TI - Use of boron waste as an additive in red bricks. AB - In boron mining and processing operations, large amounts of clay containing tailings have to be discarded. Being rich in boron, the tailings do not only cause economical loss but also pose serious environmental problems. Large areas have to be allocated for waste disposal. In order to alleviate this problem, the possibility of using clayey tailings from a borax concentrator in red brick manufacturing was investigated. Up to 30% by weight tailings addition was found to improve the brick quality. PMID- 14761762 TI - Analysis of survivors more than 10 years after heart transplantation in the cyclosporine era: Stanford experience. AB - BACKGROUND: Truly long term survival post heart transplantation has become increasingly frequent over the past two decades. METHODS: We analyzed multiple clinical outcomes in the cohort of 140 patients in the Stanford database who underwent heart transplantation after the introduction of cyclosporine-based immunosuppression in 1980 and survived >10 years after transplantation. RESULTS: We found generally excellent functional status in these patients, but a high incidence of hypertension, renal dysfunction, and graft CAD as well as malignancy. CONCLUSION: With continued improvement in post-transplant survival rates, providing complex care for such long-term recipients as these will assume increasing clinical importance in the everyday practice of transplant medicine and these data highlight the problems to be anticipated. PMID- 14761764 TI - Abdominal aortic aneurysms after heart transplantation. AB - BACKGROUND: In recent years abdominal aortic aneurysms were diagnosed in several heart transplant recipients at our center. Only case reports or small series have been reported previously and little is known about abdominal aortic aneurysms after heart transplantation. Therefore, the goals of this study were to estimate the incidence of this condition after heart transplantation, to identify risk factors for its development, and to assess its clinical consequences. METHODS: Our investigation was a retrospective, single-center cohort study of 368 consecutive patients transplanted between 1984 and 1999. RESULTS: During a mean follow-up of 75 +/- 49 months, 37 of the 368 (10%) transplant recipients and 36 of 202 (18%) of the sub-group with a history of ischemic heart disease were found to have an abdominal aortic aneurysm. All patients were male, and all except 1 had a history of ischemic heart disease. A history of ischemic heart disease prior to heart transplantation was the sole independent risk factor for developing an aneurysm by multivariate analysis. Aneurysm-related events occurred earlier and more frequently in the 7 transplant recipients who already had a dilated abdominal aorta prior to transplantation. The abdominal aortic aneurysm was the direct or indirect cause of death in at least 9 patients. CONCLUSIONS: Abdominal aortic aneurysms are relatively frequent after heart transplantation, occur at a younger age than in the general population, and have serious clinical consequences. Close ultrasonographic follow-up of patients with a history of ischemic heart disease or with an abnormal abdominal aorta prior to heart transplantation seems indicated. PMID- 14761763 TI - Bacterial mediastinitis after heart transplantation: clinical presentation, risk factors and treatment. AB - BACKGROUND: The incidence of mediastinitis after heart transplantation has been reported to be between 2.5% and 7.5%. Most previous reports from the transplant literature have assessed patients who had not received induction therapy. METHODS: From December 1996 to January 2002, a total of 230 heart transplants were performed using induction therapy with rabbit anti-thymocyte globulin at La Pitie Salpetriere Hospital (Paris, France). Mediastinitis developed in 15 patients (6.5%). A case-control study was performed to characterize the clinical presentation, microbiology, risk factors and therapy of mediastinitis after heart transplantation. RESULTS: Only 4 patients (26%) had a temperature of >38 degrees C and 6 patients (40%) had a white blood cell count of >10,000 cells/mm(3). Septicemia (46%) and positive temporary epicardial pacing wires culture (60%) were frequently observed. Staphylococcus aureus (5 of 15), Staphylococcus epidermidis (5 of 15) and gram-negative bacteria (5 of 15) were the causative organisms cultured intra-operatively. Mean duration of mechanical ventilation (2.4 vs 1.6 days; p < 0.03) and use of ventricular assistance (20% vs 0%; p < 0.04) were different between cases and controls. The mortality rate at hospital discharge was 6.7% (1 of 15). CONCLUSIONS: In the context of immunosuppression after heart transplantation, a high degree of suspicion is necessary to make the diagnosis of mediastinitis. Positive blood and temporary epicardial pacing wires cultures can be helpful in suggesting the presence of mediastinitis. Using vancomycin and an aminoglycoside as prophylaxis has to be considered because of the high prevalence of methilcilin-resistant S epidermidis and gram-negative bacteria. Conservative therapy (sternal debridement without muscle flap closure, and closed-chest drainage) showed excellent results in this series. PMID- 14761765 TI - Risk factors for recurrent rejection in pediatric heart transplantation: a multicenter experience. AB - BACKGROUND: Cardiac allograft rejection remains as a primary cause of death in the first 3 years after pediatric heart transplantation. Multiple episode of acute rejection in adult heart transplant recipients may accelerate the development of graft vasculopathy. We sought to quantify the time-related probability of recurrent rejection and identify risk factors for the development of recurrent rejection. METHODS: We analyzed data from 847 pediatric recipients who underwent transplantation between January 1, 1993 and December 31, 1998 at 22 centers in the Pediatric Heart Transplant Study (PHTS). Recurrent rejection and risk factors were evaluated using univariate and multivariate analyses. RESULTS: Five hundred fifty two patients had 1,072 rejection events and were the subject of the analyses. The highest risk of recurrent rejection occurs within 1 month after resolution of a previous rejection episode. Risk factors for recurrent rejection include the number of previous rejection events, the elapsed time since a previous rejection episode, and subjects of either Hispanic or African-American descent. Rejection associated with hemodynamic compromise and late rejection is associated with higher mortality. CONCLUSIONS: Recurrent rejection is a risk factor for mortality, especially in the presence of hemodynamic compromise. This association appears independent of the time post-transplantation. Use of surveillance biopsies appears warranted throughout the life of the transplant individual. Retransplantation should be considered among these subjects with recurrent rejection. PMID- 14761766 TI - In vitro characterization of aortic retrograde and antegrade flow from pulsatile and non-pulsatile ventricular assist devices. AB - BACKGROUND: Many advances have been made in left ventricular assist device (LVAD) development including the introduction of smaller, non-pulsatile pumps. However, controversy exists over the potential significance of non-pulsatile blood flow. In addition, some newer LVADs incorporate descending aortic anastomosis (and therefore retrograde ascending aortic flow) for outflow rather than the traditional ascending aortic anastomosis. This, combined with non-pulsatile flow, may significantly increase the risks of ascending aortic thrombus formation, especially if native cardiac function is negligible and the aortic valve remains closed. The purpose of this study was to compare pulsatile and non-pulsatile flow generated by LVADs with outflow to the ascending aorta and descending aorta. METHODS: An in vitro mock circulatory loop, driven by either a pulsatile or a non pulsatile LVAD, was anastomosed to transparent aortic models at either the ascending or descending aortic position. The aortic valve was kept closed, modeling no native cardiac output. Normal saline was used as a blood analog. Methylene blue dye was injected into the ascending aorta and aortic arch to demonstrate flow patterns. Dye washout time (in seconds) was used as a marker of flow stagnation and potential thrombogenicity. LVAD flow, rate, after-load and coronary flow were measured. RESULTS: Dye washout times at a flow rate of 5 liters/min were 1.7 +/- 0.75, 2.1 +/- 0.71, 4.7 +/- 0.82 and 9.9 +/- 4.4 seconds for pulsatile ascending (PA), non-pulsatile ascending (NPA), pulsatile descending (PD) and non-pulsatile descending flow (NPD), respectively. Coronary flow averaged 294 ml/min over all set-ups. Dye washout times at a flow rate of 4 liters/min were 3.0 +/- 1.0, 3.0 +/- 0.8, 14.0 +/- 3.8 and 25.0 +/- 9.1 seconds for PA, NPA, PD and NPD, respectively. Coronary flow averaged 227 ml/min over all set-ups. Ascending aortic anastomoses were associated with shorter dye washout times compared with descending aortic anastomoses, regardless of flow type (p < 0.001). There was no difference in washout time between pulsatile and non pulsatile flow in the ascending aortic position (p = 0.23 and 0.12 for 5 and 4 liters/min, respectively). Pulsatile flow in the descending aorta had shorter washout times than non-pulsatile flow in the descending aorta (p < 0.001 and p = 0.004 for 5 and 4 liters/min, respectively). CONCLUSIONS: LVAD descending aortic anastomosis and retrograde aortic flow is associated with increased flow stagnation in the ascending aorta. This may increase the risk for thrombus formation in patients relying solely on retrograde aortic flow, especially if cardiac function and antegrade blood flow returns. PMID- 14761767 TI - Cyclosporine treatment preserves coronary resistance artery function in rat cardiac allografts. AB - BACKGROUND: A marked decline in vascular myogenic response occurs during the course of rat cardiac allograft rejection. Two important contributory features are an inducible nitrous oxide synthase (iNOS)-catalyzed, NO-mediated vasodilation and a loss of smooth muscle function. In this study, we examine the effect of cyclosporine immunosuppressive therapy on the alleviation of arterial dysfunction of coronary resistance arteries in allografts using pressure myography. METHODS: Rats receiving heterotopic abdominal cardiac transplantation were treated with cyclosporine (5 mg/kg), Cremophore or distilled water. Coronary septal arteries (internal diameter 200 microm) were dissected from isograft (Lewis to Lewis) and allograft (Fisher to Lewis) rat hearts at Day 21 post transplantation and mounted on a pressure myograph. Pressure-induced vasoconstriction was measured before and after iNOS inhibition with aminoguanidine (AG; 100 micromol/liter). Both endothelium-based (ACh-induced) and endothelium-independent (sodium nitroprusside-induced) vasorelaxation were also recorded in each group. RESULTS: Pressure-induced myogenic contraction was reduced in allograft coronary arteries at Day 21 post-transplantation compared with matched isografts (p < 0.05). AG potentiated myogenic tone in allograft arteries, but had no effect on untreated Day 21 isograft vessels, indicating the presence of iNOS-based relaxation only in allograft vessels. Depolarization induced vasoconstriction was lower in allograft compared with isograft arteries (p < 0.05). Cyclosporine therapy also improved depolarization-induced constriction in allograft vessels compared with untreated groups (p < 0.05). Furthermore, cyclosporine therapy preserved endothelium-based and endothelium independent vasorelaxation in allograft arteries at Day 21 post-transplantation. CONCLUSIONS: Cyclosporine immunosuppressive therapy has a significant effect on the alleviation of early endothelial and smooth muscle dysfunction in coronary allograft arteries. PMID- 14761768 TI - Pre- and post-transplant anti-myosin and anti-heat shock protein antibodies and cardiac transplant outcome. AB - BACKGROUND: The purpose this study was to investigate the relationship of anti myosin and anti-heat shock protein immunoglobulin G (IgG) serum antibodies to the original heart disease of cardiac transplant recipients, and also to rejection and patient survival after cardiac transplantation. METHODS: Anti-myosin and anti heat shock protein (anti-hsp) IgG antibodies were evaluated in pre-transplant sera from 41 adult cardiac allograft recipients and in sequential post-transplant serum samples from 11 recipients, collected at the time of routine endomyocardial biopsies during the first 6 months after transplantation. In addition, the levels of these antibodies were determined from the sera of 28 healthy blood donors. RESULTS: Higher anti-myosin antibody levels were observed in pre-transplant sera than in sera from normal controls. Moreover, patients with chronic Chagas heart disease showed higher anti-myosin levels than patients with ischemic heart disease, and also higher levels, although not statistically significant, than patients with dilated cardiomyopathy. Higher anti-hsp levels were also observed in patients compared with healthy controls, but no significant differences were detected among the different types of heart diseases. Higher pre-transplant anti myosin, but not anti-hsp, levels were associated with lower 2-year post transplant survival. In the post-transplant period, higher anti-myosin IgG levels were detected in sera collected during acute rejection than in sera collected during the rejection-free period, whereas anti-hsp IgG levels showed no difference between these periods. CONCLUSIONS: The present findings are of interest for post-transplant management and, in addition, suggest a pathogenic role for anti-myosin antibodies in cardiac transplant rejection, as has been proposed in experimental models of cardiac transplantation. PMID- 14761769 TI - Detection of regulatory cells as an assay for allograft tolerance in miniature swine. AB - BACKGROUND: There is currently a great need for an in vitro assay to assess the presence of tolerance following allotransplantation to determine whether immunosuppressive medications can be discontinued. Our laboratory has recently developed an assay involving coculture inhibition of cell-mediated lympholysis that correlates with tolerance to allografts in swine leukocyte antigen (SLA) Class I-mismatched miniature swine. The potential for clinical application of this assay may depend on 2 important factors: (1) whether the assay can be used in the presence of immunosuppression; and (2) whether frozen-stored naive responder cells can be utilized. METHODS: Long-term tolerant MGH miniature swine that had accepted SLA Class I-mismatched kidney transplants after a 12-day course of cyclosporine or tacrolimus were studied. Two long-term tolerant and 2 naive control animals were treated with a clinically relevant dose of cyclosporine for 2 weeks (trough level 100 to 400 ng/ml) to simulate the ongoing "chronic" immunosuppression used in human recipients of allografts. Cells from tolerant or naive, recipient-matched animals were stimulated for 6 days with donor or third party SLA. These primed cells were then cocultured with naive unstimulated recipient major histocompatibility complex (MHC)-matched responders and irradiated stimulators. Responder cells were tested both fresh and frozen. RESULTS: Suppression of cytotoxic responses of naive responder cells was observed in all coculture assays using cells from tolerant animals primed against donor antigen in vitro, but not in assays using similarly primed cells from naive animals. Responder cells from tolerant animals receiving immunosuppression had a suppressive activity similar to that from cells of the same animals not receiving immunosuppression. Similar suppression was also observed in coculture assays using either fresh or frozen naive responder cells. CONCLUSIONS: This coculture assay appears to correlate with the presence of tolerance under conditions applicable to the clinical setting. The assay appears to identify peripheral regulatory mechanisms of tolerance in allogeneic transplant recipients, and therefore may provide an approach for determining an appropriate timepoint at which to test withdrawal of immunosuppressive medications. PMID- 14761770 TI - Increased matrix metalloproteinase-2 and membrane type 1 matrix metalloproteinase activity and expression in heterotopically transplanted murine tracheas. AB - BACKGROUND: Bronchiolitis obliterans syndrome (BOS) is the most common long-term cause of morbidity and mortality after heart-lung or lung transplantation. One pathologic feature of BOS is infiltration of fibroblasts and connective tissue products into the airway lumen, which form a fibrous, collagen-rich occlusion. Heterotopically transplanted allogeneic murine tracheal stenosis resemble BOS in the development of obliterans airway disease. Matrix metalloproteinases (MMPs) are key enzymes involved in tissue remodeling and, clinically, have several roles in pulmonary diseases. Among the MMP family, type IV collagenases, MMP-2 and MMP 9, have high gelatinolytic activity and are thought to play a role in several pulmonary diseases. Membrane type 1 MMP (MT1-MMP) activates the zymogen of MMP-2 (proMMP-2, 72 kd), and activated MMP-2 (active MMP-2, 62 kd) degrades type IV collagen and plays an important role in clinical pulmonary disease. In this study, we examine the expression of MMP-2, its activator MT1-MMP and MMP-9 in BOS using murine trachea transplantation models. METHODS: Rats were divided into 5 experimental groups (n = 10 in each group). Group I was a control group with intact tracheas. Animals with tracheal grafts underwent heterotopically syngeneic (Groups II and III) or allogeneic (Groups IV and V) transplantation. The recipient rats were killed 7 days (Groups II and IV) or 28 days (Groups III and V) after transplantation. The harvested tracheal grafts were examined histologically. MMP activity was assessed using gelatin zymography analysis, and MMP-2 and MT1-MMP gene expression was examined by quantitative real-time polymerase chain reaction analysis. Distribution of gelatinolytic activity was studied using in situ zymography. RESULTS: There was little histologic change in the intact trachea (Group I) and in all isografts (Groups II and III). Fibrotic tissues in Group V significantly occluded the tracheal lumen, and there was severe lymphocyte infiltration in Group IV. According to gelatin zymography, proMMP-9 was faint at 7 days, but activated MMP-9 was not present in all groups. The MMP-2 gelatinolytic bands were predominant; the activation in Group V was significantly greater than that in Group IV, and in Group III it was significantly greater than that in Group II. Gene expression of both MMP-2 and MT1-MMP were significantly higher in Group V than in the other groups (p < 0.01), and MMP-2 was clearly activated. Gelatinolytic activity was localized in the fibrotic tissues or lymphocytes of thickening lumen after destruction of the epithelium by stenosis. CONCLUSIONS: These results demonstrate that MMP-2, together with its activator MT1-MMP, may have an important role in the development of BOS, which is associated with destruction of the tracheal epithelium, leading to fibrosis. PMID- 14761771 TI - Increased toll-like receptor 4 in the myocardium of patients requiring left ventricular assist devices. AB - BACKGROUND: Cytokine activation in the myocardium of deteriorating patients with heart failure who undergo left ventricular assist-device (LVAD) implantation has been documented, but the underlying mechanisms remain poorly understood. We hypothesized the innate immune system is activated with expression of Toll-like receptor 4 (TLR4), leading to cytokine activation in these patients. METHODS: We used quantitative real-time reverse-transcriptase polymerase chain reaction to measure TLR4, interleukin-1 (IL-1) receptor, IL-1 beta, IL-6, and tumor necrosis factor alpha (TNF-alpha) mRNA expression in myocardial samples from 36 patients. We compared 18 patients who underwent LVAD implantation with 18 patients with less severe heart failure who underwent elective heart transplantation. RESULTS: Toll-like receptor 4 expression was 1.69-fold greater (p < 0.05) and IL-1 receptor expression was 3.64-fold greater (p < 0.0001) in the deteriorating patients who required LVADs. Myocardial TNF-alpha (1.71-fold, p < 0.05), IL-6 (2.57-fold, p < 0.005), and IL-1 beta (9.78-fold, p < 0.001) also were increased in the LVAD candidates. Toll-like receptor 4 expression correlated strongly with IL-1 receptor expression (r= 0.75, p < 0.0001) and with IL-1 beta expression in individual patients (r = 0.7, p < 0.0001). Interleukin-1 receptor expression also correlated with IL-1 beta expression (r = 0.78, p < 0.0001) within patients. We found no correlation between TLR4 and either TNF-alpha or IL-6 expression. CONCLUSIONS: Patients who required LVAD support showed evidence of innate immune system activation, indicated by an increase in the key effector molecule TLR4 associated with a specific pattern of cytokine expression in the myocardium. PMID- 14761772 TI - Right ventricle-sparing heart transplantation effective against iatrogenic pulmonary hypertension. AB - BACKGROUND: Right heart failure is the predominant cause of death following heart transplantation, occurring with disturbingly high frequency in patients with severe antecedent pulmonary hypertension. We have recently reported a novel technique of heart transplantation that spares the recipient right ventricle, excising only the recipient left ventricle. The resulting model has 2 right hearts and 1 left heart. The aim is to preserve the recipient's right ventricle, which is already conditioned to pulmonary hypertension. The hope is that, in this way, death due to right heart failure can be prevented in humans. Our prior report was a feasibility study in normal dogs. This study challenges this new technique by creating iatrogenic pulmonary hypertension in the recipient animals. METHODS: Iatrogenic pulmonary hypertension was created in 4 recipient canines by intravenous injection of the pulmonary toxin monocrotaline pyrrole (single bolus of 3.5 to 4.5 mg/kg intravenously [i.v.]). RESULTS: Within 6 weeks of monocrotaline administration, relative pulmonary hypertension occurred (mean pulmonary artery [PA] pressure 20 mm Hg vs 10 mm Hg for controls [p < 0.01]) (pulmonary vascular resistance [PVR] 4.2 vs 1.5 Wood units [P < 0.01]), and right ventricular (RV) hypertrophy developed (RV thickness 11 mm vs 2 mm [P < 0.04]). Histologic examination confirmed severe muscle infiltration and thickening of the media of the pulmonary arterioles. RV-sparing heart transplantation was performed successfully in all 4 animals with pulmonary hypertension. In all cases, the animals were weaned without difficulty from cardiopulmonary bypass, despite the ambient pulmonary hypertension, on low-dose epinephrine, maintaining systolic blood pressure of 104 mm Hg at right atrial pressure of 7 mm Hg. Both right hearts contracted well without dilation or strain. A single "control" traditional orthotopic transplant experiment in an animal with monocrotaline-induced pulmonary hypertension resulted in immediate death from right heart failure. CONCLUSIONS: Right ventricle-sparing heart transplantation ("one-and-one-half heart model") can handle pulmonary hypertension without difficulty. This evidence adds impetus for further pursuing of right ventricle-sparing heart transplantation to decrease the incidence of death from right heart failure in recipients with severe antecedent pulmonary hypertension. PMID- 14761774 TI - Heart transplantation in a patient with chloroquine-induced cardiomyopathy. AB - We present the first report of a patient who underwent heart transplantation (HT) after endomyocardial biopsy (EMB) and revealed chloroquine-induced cardiomyopathy (CIC). This patient, who was treated with chloroquine for 6 years, developed a restrictive cardiomyopathy that progressed to congestive heart failure (CHF) resistant to medical management. PMID- 14761773 TI - Partial liquid ventilation with perfluorodecalin following unilateral canine lung allotransplantation in non-heart-beating donors. AB - BACKGROUND: The purpose of this study was to evaluate canine lungs obtained from non-heart-beating donors after unilateral lung transplantation subjected to partial liquid ventilation with perfluorodecalin. METHODS: Twelve donor dogs were killed and kept under mechanical ventilation for 3 hours. Heart-lung blocks were harvested after retrograde pulmonary hypothermic flush with Perfadex. Left lung grafts were randomly transplanted into 12 weight-matched recipient animals. Animals were divided into 2 groups: control (standard mechanical ventilation, n = 6) and PLV (partial liquid ventilation, n = 6). Forty-five minutes after transplantation, the animals in the PLV group received perfluorodecalin (15 ml/kg) via orotracheal tube. All animals received volume-controlled ventilation (FIO2) 1.0, PEEP 5 cm H(2)O) over 6 consecutive hours. Thereafter, blood-gas analysis, ventilatory mechanics and hemodynamics were registered at 30-minute intervals. After 6 hours of reperfusion the animals were killed and the transplanted lungs were extracted to obtain the wet/dry weight ratio. RESULTS: There were significant differences in pulmonary arterial pressure, which were higher in control group animals (p < 0.009). The control animals also showed higher arterial PaO(2) than those in the PLV group (p < 0.00001), but lower PaCO(2) (p < 0.008). The peak and plateau pressures were higher in the PLV group (p < 0.00001). Neither static compliance nor wet/dry weight ratios were different in between groups. CONCLUSIONS: PLV with perfluorodecalin yields functional results compatible with life in this model. Nonetheless, pulmonary gas exchange and mechanics were superior after reperfusion in animals given conventional mechanical ventilation up to 6 hours after left lung allotransplantation. PMID- 14761775 TI - Serial dobutamine stress echocardiography with Doppler assessment of the left internal thoracic artery graft after minimally invasive bypass for a patient with an orthotopic heart transplant. AB - Select patients who have undergone orthotopic heart transplantation with proximal left anterior disease may be candidates for minimally invasive direct coronary artery bypass surgery. Combining left internal thoracic artery transthoracic Doppler flow assessment with wall motion assessment during dobutamine stress echocardiography adds to the utility of this test by focusing attention on the graft's status as well as detecting ischemia due to cardiac allograft vasculopathy. PMID- 14761776 TI - Dobutamine-related asthma in a patient awaiting cardiac transplantation: the eosinophilic dilemma. AB - Long-term continuous intravenous inotrope infusion is frequently used as a pharmacologic bridge to cardiac transplantation in patients with end-stage congestive heart failure. We report a case of severe eosinophil-associated asthma after 6 weeks of therapy with the inotrope dobutamine in a patient awaiting cardiac transplantation. Review of the laboratory data revealed a steady increase in eosinophils during the period of treatment with dobutamine. Once the patient was switched to an alternative inotrope, milrinone, the eosinophil count returned to normal, and the asthma exacerbation resolved. PMID- 14761779 TI - Mechanism of dystrophic calcification in rabbit aortas: temporal and spatial distributions of calcifying vesicles and calcification-related structural proteins. AB - INTRODUCTION: We have previously demonstrated the accumulation of calcifying vesicles in the thoracic aorta undergoing dystrophic calcification. Whether the distributions of other structural proteins related to calcification such as collagen and elastin fibers undergo coordinated modifications has not been established. METHODS: Young rabbits with various degrees of aortic calcification induced by cholesterol dietary interventions were used as an animal model to study the correlations. RESULTS: Rabbits fed a diet enriched in cholesterol for 3 months accumulated calcifying vesicles in the ascending thoracic aortas but did not develop histologically identifiable calcification. There were concomitant marked thickenings of the intima with focal deposition of collagen and disruption of the internal elastic fibers at this stage. By the 6th month, calcification was predominantly present in the intimal area adjacent to the media. At this calcified stage, calcifying activity of vesicles was higher than earlier stages. Concomitantly, collagen deposition in the lesions was intensified and the internal elastic fibers were completely disintegrated. These changes were found to be more profound in the proximal than in the distal portion of the aortas. CONCLUSION: The coordinated changes in the accumulation of collagen, disintegration of internal elastic fibers, and the appearance of calcifying vesicles in the lesions before calcification may set the stage for aortic calcification. PMID- 14761780 TI - Myocardial hypertrophy and enhanced left ventricular contractility in Zucker diabetic fatty rats. AB - Heart failure is known to be a complication of insulin-dependent (IDDM) and noninsulin-dependent diabetes mellitus (NIDDM) even in the absence of coronary heart disease or hypertension. The mechanisms leading to diabetic cardiomyopathy are unknown. The aim of the study was to characterize structural and functional alterations in hyperinsulinemic Zucker diabetic fatty (ZDF) rats treated with or without insulin. Diabetic animals showed a twofold increase in cardiomyocyte volume with increased left ventricular ANP but not BNP mRNA levels in spite of a reduced plasma renin activity (PRA) 2 months after onset of diabetes compared to nondiabetic littermates. These changes were associated with an increase in left ventricular performance as assessed by echocardiography. Insulin treatment led to a significant increase in body weight (BW), total heart weight, myocardial protein content, and left ventricular mass (LVM). Perivascular fibrosis and laminin thickness were significantly augmented in diabetic rat myocardium irrespective of insulin treatment, whereas interstitial collagen I and fibronectin were similarly found in diabetic and control myocardium. Initial stages of diabetic cardiomyopathy in hyperinsulinemic rats are characterized by cardiomyocyte hypertrophy and enhanced cardiac contractility. It is suggested that hyperinsulinemia may be involved in cardiac hypertrophy. PMID- 14761781 TI - Intimal hyperplasia in the distal ulnar artery; Influence of gender and implications for the hypothenar hammer syndrome. AB - INTRODUCTION: The distal ulnar artery in the hand is subjected to trauma when the hypothenar eminence is used as a hammer. The clinical presentation of arterial obstruction or aneurysm formation at this site with a history of trauma to the hypothenar eminence is termed the hypothenar hammer syndrome (HHS) and is most commonly observed in the right hands of men. Since the trauma-inducing activities associated with the HHS are common, the histopathology of this arterial segment in our autopsy population was evaluated. METHODS: The distal ulnar artery in the hand was examined in 21 randomly selected autopsies. Formalin-fixed paraffin embedded cross sections of artery were assessed using hematoxylin and eosin (H&E), trichrome and elastic stains as well as immunohistochemistry for CD34 and alpha-smooth muscle actin. RESULTS: Intimal hyperplasia is common at this site in the vasculature. The degree of intimal hyperplasia, as assessed by the intima/media ratio, is significantly greater in the right hands of men than in either their corresponding left hands or in women. None of the arterial segments examined contained foam cells or extracellular lipid deposits indicative of atherosclerosis. CONCLUSIONS: In the injury-prone segment of the distal ulnar artery, intimal hyperplasia is common but is particularly severe in the right hands of men, correlating with the demographics of the HHS. Although, this arterial segment frequently develops intimal hyperplasia, it is resistant to the development of atherosclerosis. PMID- 14761782 TI - Structural and molecular pathology of the heart in Carvajal syndrome. AB - BACKGROUND: Carvajal syndrome is a familial cardiocutaneous syndrome consisting of woolly hair, palmoplantar keratoderma, and heart disease. It is caused by a recessive deletion mutation in desmoplakin, an intracellular protein that links desmosomal adhesion molecules to intermediate filaments of the cytoskeleton. The pathology of Carvajal syndrome has not been described. METHODS: Here, we report the first description of the structural and molecular pathology of the heart in Carvajal syndrome. We characterized gross and microscopic pathology and identified changes in expression and distribution of intercalated disk and intermediate filament proteins in ventricular myocardium. RESULTS: We identified a unique cardiomyopathy characterized by ventricular hypertrophy and dilatation, focal ventricular aneurysms, and distinct ultrastructural abnormalities of intercalated disks, but no evidence of fibrofatty infiltration or replacement of myocardium. We also observed markedly decreased amounts of specific immunoreactive signal for desmoplakin, plakoglobin, and the gap junction protein, connexin43, at intercalated disks. The intermediate filament protein, desmin, which is known to bind desmoplakin, showed a normal intracellular pattern of distribution but failed to localize at intercalated disks. CONCLUSIONS: The desmoplakin mutation in Carvajal syndrome produces a cardiomyopathy with unique pathologic features. Altered protein-protein interactions at intercalated disks likely cause both contractile and electrical dysfunction in Carvajal syndrome. PMID- 14761783 TI - Allograft arteriopathy: pathogenesis update. AB - Allograft arteriopathy is a vascular intimal proliferative process that occurs in all solid organ transplants and stands as the single most significant obstacle to successful long-term solid organ transplantation; it shares a number of pathologic features with restenosis lesions and atherosclerosis. This article will review some of the newer developments in our understanding of the immunological and vascular biology underpinnings of the disease, including the roles played by cytokine and chemokine mediators in recruiting and activating both inflammatory cells, as well as smooth muscle cell precursors. PMID- 14761784 TI - Cerivastatin-induced apoptosis of human aortic smooth muscle cells through partial inhibition of basal activation of extracellular signal-regulated kinases. AB - BACKGROUND: The beneficial effects of cerivastatin including hypolipidemic properties have been demonstrated to involve nonlipid as well as lipid mechanisms. In the present study, we examined the mechanisms underlying cerivastatin-induced growth inhibition of human aortic smooth muscle (ASM) cells. METHODS: Human ASM cells were cultured in 96-well plates with or without cerivastatin in the presence or absence of mitogen-activated protein (MAP) kinase (MAPK)/extracellular signal-regulated kinase (ERK) kinase MEK1/MEK 2 inhibitor PD98059. Cell growth was assessed by colorimetric quantitation of NADH, and cell viability was determined by trypan blue dye exclusion method. The induction of apoptosis was determined by propidium iodide (PI) staining method using flow cytometer. The activation of ERKs or c-Jun N-terminal kinases (JNKs) was determined by Western blotting using antibodies (Abs) specific for phospho-ERKs or phospho-JNKs. RESULTS: Treatment of the ASM cells with cerivastatin prevented cell growth in a concentration-dependent manner through at least induction of apoptosis. The cerivastatin-induced apoptosis was reversed by coincubation with isoprenoid [mevalonate, geranylgeranyl pyrophosphate (GGPP), and farnesyl pyrophosphate (FPP)] suggesting a role for isoprenoid in the cerivastatin-induced apoptosis. The cerivastatin cooperated with a MEK1/MEK2 inhibitor PD98059 to induce apoptosis, which appeared to correlate with down-regulation of ERK activation (phospho-ERKs expression) induced by the combination. CONCLUSION: Cerivastatin-induced blockade of ERK activation in ASM cells might result in growth inhibition including apoptosis, which might explain some aspects of the beneficial effects of cerivastatin on coronary artery disease. PMID- 14761785 TI - Cardiac plasmacytoma. AB - A 69-year-old white woman presented with a 3-month history of progressive dyspnea, orthopnea, fatigue and weakness. Clinical, diagnostic imaging and echocardiographic investigations suggested an occult primary cancer with metastasis to the heart. The patient's condition deteriorated gradually, and she died 2 months later. At autopsy, a malignant tumor encasing the heart and a 1-cm solitary tumor nodule in the lower lobe of the left lung were found. Histologic and electron microscopic studies revealed a plasmacytoma predominantly involving the epicardium and a small solitary plasmacytoma located in the left lung. The two tumors were further confirmed by immunohistochemical studies that showed monoclonal IgG expression and kappa light chain restriction. PMID- 14761786 TI - Coarctation of the aorta in the adult complicated by poststenotic fusiform aneurysm. AB - We describe a case of adult coarctation of the aorta, which presented unusually as a poststenotic fusiform aneurysm in a 48-year-old female. She was normotensive and had a history of back pain and palpitations. Under partial cardiopulmonary bypass, the stenosis and the aneurysm were resected and replaced with a Dacron tube graft. PMID- 14761787 TI - Cardiac thromboemboli complicating a stab wound to the heart. AB - A 27-year-old man was noted to have neurologic deficit 4 days following cardiorrhaphy to repair a penetrating cardiac injury. Cerebral computed tomography scan showed multiple embolic infarcts and two-dimensional echocardiography revealed the source as thrombus in the left ventricle. Although this entity has been described following blunt chest trauma and cardiac contusion, it has not been noted in association with penetrating injury. Intracardiac thrombus with systemic emboli should be considered as a possible complication following cardiorrhaphy for penetrating trauma. PMID- 14761788 TI - Cryopreserved pulmonary homograft: postimplant changes. PMID- 14761789 TI - Comparative sequence analysis of Sordaria macrospora and Neurospora crassa as a means to improve genome annotation. AB - One of the most challenging parts of large scale sequencing projects is the identification of functional elements encoded in a genome. Recently, studies of genomes of up to six different Saccharomyces species have demonstrated that a comparative analysis of genome sequences from closely related species is a powerful approach to identify open reading frames and other functional regions within genomes [Science 301 (2003) 71, Nature 423 (2003) 241]. Here, we present a comparison of selected sequences from Sordaria macrospora to their corresponding Neurospora crassa orthologous regions. Our analysis indicates that due to the high degree of sequence similarity and conservation of overall genomic organization, S. macrospora sequence information can be used to simplify the annotation of the N. crassa genome. PMID- 14761790 TI - Image analysis of hyphal morphogenesis in Saprolegniaceae (Oomycetes). AB - Because of their wide range of apical morphology, several members of saprolegniaceous fungi (Oomycetes) were chosen to examine concordance with the vesicle supply center (VSC) model of hyphal morphogenesis. Two computer routines were devised to measure diameter changes over long stretches of hyphae and to test compatibility with the theoretical hyphoid shape, y = xcot(xV/N). In all four genera examined, the apex followed closely the contour described by the hyphoid equation; divergences became evident in the subapex. The hyphae of Saprolegnia parasitica showed maximum concordance with the VSC model, i.e., their profile matched a hyphoid curve from the apex to the entire length of the mature hyphal tube. In Aphanomyces and Leptolegnia, growth in the subapical region subsided becoming less than that specified by the hyphoid equation. In Achlya bisexualis, the reverse was true, the subapical region expanded beyond that specified by the hyphoid equation. The two divergent subapical tendencies gave the hyphal tips a cylindroid or conoid appearance, respectively. Since the hyphal apex of all four species conformed to the curvature dictated by the hyphoid equation, we concluded that a basic VSC mechanism operates in all of these oomycetous fungi. Accordingly, we suggest that the shape of an oomycetous hypha is generated by a VSC-driven gradient of wall formation, which is subject to additional modification in the subapex to produce a range of hyphal tip morphologies. The mathematical basis for generating a conoid hyphal tip by elongating the VSC is described in Appendix A. PMID- 14761791 TI - MATE transposable elements in Aspergillus nidulans: evidence of repeat-induced point mutation. AB - The sequences of five MATE transposable elements were retrieved from the Aspergillus nidulans genome sequence. These elements are 6.1 kb in length and are characterized by 9-10 bp target site duplications, paired approximately 40 bp palindromes close to each end, and in the unmutated elements, 57 clustered Spe motifs (RWCTAGWY) scattered through their length. Short open reading frames have no known homology. Two of the MATE elements have numerous C --> T transitions on both DNA strands relative to the remaining three elements. These mutations have all the characteristics of repeat-induced point mutation (RIP) previously described in Neurospora crassa, but not experimentally demonstrated in A. nidulans. Ninety-eight percent of mutated cytosines are in CpG and CpA doublets, the former mutating at higher frequency. PMID- 14761792 TI - cAMP regulation of "pathogenic" and "saprophytic" fungal spore germination. AB - We report on the elucidation of two separate pathways of spore germination in a plant pathogenic fungus Colletotrichum gloeosporioides f. sp. aeschynomene. Conidia of the fungus can germinate either from one side or from both sides, depending on external conditions. In shake culture that includes an extract made up from fresh peas, the unicellular conidium divides and one of the two cells develops a germ tube. On a solid surface this germ tube differentiates an appressorium. In rich medium without pea extract, germination is highly similar to Aspergillus spore germination: the conidium swells, forms a single germ tube and then divides and forms a second germ tube. Conidia that germinate in a rich medium do not form appressoria even on a solid surface and are non-pathogenic. In rich medium, cAMP stimulates germination in rich liquid cultures and induces appressoria formation on a hard surface. In pea extract cAMP induces swelling and formation of irregular germ tubes and appressoria. Our results suggest that plant surface signals induce pathogenic-specific spore germination in a cAMP independent manner. cAMP is required for saprophytic germination and for appressorium formation. PMID- 14761793 TI - Proteomic analysis of rutin-induced secreted proteins from Aspergillus flavus. AB - Few studies have been conducted to identify the extracellular proteins and enzymes secreted by filamentous fungi, particularly with respect to dispensable metabolic pathways. Proteomic analysis has proven to be the most powerful method for identification of proteins in complex mixtures and is suitable for the study of the alteration of protein expression under different environmental conditions. The filamentous fungus Aspergillus flavus can degrade the flavonoid rutin as the only source of carbon via an extracellular enzyme system. In this study, a proteomic analysis was used to differentiate and identify the extracellular rutin induced and non-induced proteins secreted by A. flavus. The secreted proteins were analyzed by two-dimensional electrophoresis and MALDI-TOF mass spectrometry. While 15 rutin-induced proteins and 7 non-induced proteins were identified, more than 90 protein spots remain unidentified, indicating that these proteins are either novel proteins or proteins that have not yet been sequenced. PMID- 14761794 TI - Functional analysis of tvsp1, a serine protease-encoding gene in the biocontrol agent Trichoderma virens. AB - Serine proteases are highly conserved among fungi and considered to play a key role in different aspects of fungal biology. These proteases can be involved in development and have been related to pathogenesis or biocontrol processes. A gene (tvsp1) encoding an extracellular serine protease was cloned from Trichoderma virens, a biocontrol agent effective against soilborne fungal pathogens. The gene was expressed in Escherichia coli and a polyclonal antibody was raised against the recombinant protein. The expression pattern of tvsp1 was determined and its physiological role was addressed by mutational analysis. Strains of T. virens in which tvsp1 was deleted (PKO) or constitutively overexpressed (POE) were not affected in growth rate, conidiation, extracellular protein accumulation, antibiotic profiles nor in their ability to induce phytoalexins in cotton seedlings. Tvsp1 overexpression, however, significantly increased the ability of some strains to protect cotton seedlings against Rhizoctonia solani. Our data show that Tvsp1 is not necessary for the normal growth or development of T. virens, but plays a role in the biocontrol process. PMID- 14761795 TI - Gene expression during Ustilago maydis diploid filamentous growth: EST library creation and analyses. AB - Ustilago maydis is an important model system for the plant pathogenic smut and rust fungi. Critical to the continued development of this model is establishing genomic resources. We have constructed a cDNA library from a forced diploid culture of U. maydis growing as filaments and have generated 7455 ESTs that are assembled into 3074 contiguous sequences. This represents as much as 46% of the coding capacity predicted for U. maydis. BLAST searches with a similarity cutoff of E Ca.G=Ca.A. CONCLUSIONS: 8-OxoG-derived lesions in DNA such as spiroiminodihydantoin (Sp), guanidinohydantoin (Gh), oxaluric acid (Oa), oxazolone (Oz) and Ca are substrates of base excision repair DNA N-glycosylases. Most of them, Sp, Gh, Oa and Oz, are substrates of the oxidized bases-specific enzymes such as Nth or Fpg. In contrast, Ca is substrate of the human methylpurine DNA N-glycosylase (Mpg). PMID- 14761848 TI - Human and rodent cell lines showing no differences in the induction but differing in the repair kinetics of radiation-induced DNA base damage. AB - PURPOSE: To compare the induction and repair of radiation-induced base damage in human and rodent cell lines. MATERIAL AND METHODS: Experiments were performed with two human (normal fibroblasts HSF1 and tumour HeLa cells) and two rodent (mouse L929 and hamster CHO-K1) cell lines. Base damage was determined with the alkaline comet assay combined with the repair enzyme formamidopyrimidine glycosylase (Fpg). Proteins were detected by Western blot. RESULTS: The induction of Fpg-sensitive sites was measured in human and rodent cell lines for doses up to 8 or 5 Gy, respectively. Comets were analysed in terms of tail moments, which were transformed into Gy-equivalents. The amount of Fpg-sensitive sites increased linearly with doses up to 4 Gy, whereby the ratio of single-strand breaks (ssb) to Fpg-sensitive sites was nearly identical for human and rodent cells with ssb:Fpg-sensitive sites=1:0.41+/-0.07 and 1:0.45+/-0.05, respectively. For doses exceeding 4 Gy, the amount of Fpg-sensitive sites did not increase further, indicating a dose limit up to which the comet assay can be used to detect Fpg sensitive sites. Repair of Fpg-sensitive sites was studied for an X-ray dose of 4 Gy. For all four cell lines, the repair was measured to be completed 24 h after irradiation, but with pronounced differences in the kinetics. In both rodent cell lines, 50% of Fpg-sensitive sites were removed after t((1/2))=25+/-10 min in contrast to t((1/2))=80+/-20 min in the two human cell lines. The two species also differed in the level of polymerase ss with, on average, a three- to fivefold higher level in rodent cells compared with human cells. CONCLUSIONS: Repair of radiation-induced Fpg-sensitive sites was much faster in rodent than in human cells, which might result from the higher level of polymerase ss found in rodent cells. PMID- 14761849 TI - Radiation-induced apoptosis of oligodendrocytes and its association with increased ceramide and down-regulated protein kinase B/Akt activity. AB - PURPOSE: Oligodendrocytes are cells responsible for myelination in the central nervous system and have been shown to undergo radiation-induced apoptosis. The roles of ceramide and protein kinase B/Akt (PKB/Akt) were assessed in radiation induced apoptosis of oligodendrocytes in vitro. MATERIALS AND METHODS: Primary cultures of oligodendrocytes were established from neonatal rat brains and cell identity was assessed by immunohistochemistry. Apoptosis was assessed histologically according to its specific morphologic features using 4',6 diaminido-2-phenylindole, and by transferase-mediated deoxynucleotidyl transferase nick end-labelling staining. The ceramide level was measured using a diacyglycerol kinase assay, and PKB/Akt activity was determined using immunoblotting and a protein kinase assay. RESULTS: Ionizing radiation, C2 ceramide or wortmannin induced apoptosis in oligodendrocytes but not astrocytes. A rapid increase in ceramide was observed in oligodendrocytes after ionizing radiation. Monensin, an inhibitor of acid sphingomyelinase, reduced the apoptotic response in oligodendrocytes after ionizing radiation. Fumonisin B1, an inhibitor of ceramide synthase, showed no such effect in the cells. Radiation-induced apoptosis of oligodendrocytes was associated with a decrease in PKB activity, similar to that observed after treatment with C2-ceramide or wortmannin, but not after dihydro-C2-ceramide. Confocal microscopy revealed a loss of phosphorylated PKB immunostaining in the nucleus of apoptotic oligodendrocytes after ionizing radiation or C2-ceramide treatment. The level of phosphorylated FKHRL1, a transcription factor phosphorylated by PKB, decreased in irradiated oligodendrocytes. CONCLUSIONS: A ceramide-PKB-mediated signalling pathway might play a role in radiation-induced apoptosis of oligodendrocytes. PMID- 14761850 TI - Role of p53 mutations, protein function and DNA damage for the radiosensitivity of human tumour cells. AB - PURPOSE: The tumour suppressor protein p53 is considered to have an impact on the radiosensitivity of tumour cells. However, this concept does not easily translate to the tumour sensitivity in the clinics. The aim of the present study was to determine whether a functional or dysfunctional p53 is associated with a sensitive or resistant phenotype. It was further studied whether DNA damage might be an additive factor by which p53 has impact on cell survival. MATERIALS AND METHODS: Nine human tumour cell lines were studied for p53 mutation by direct sequencing of exons 4-9. Regulation of p53 and p21(cip1/waf1) protein was assessed by immunoblotting and cell cycle effects by combining 5 bromodeoxyuridine incorporation and flow cytometry. RESULTS AND CONCLUSION: Three strains (RT112, Du145, SCC4451) were found to have a missense-mutation in the core domain and one did not express p53 at all (HeLa), presumably due to HPV18 infection. Immunoblots of these cells showed neither a regulated p53 nor p21 expression. The cells did not arrest in G1 phase after X-irradiation but did arrest in G2/M. All cells expressing wild-type protein (LNCaP, T47D-B8, MCF-7 and sublines BB and Bus) showed an intact p53 and p21 regulation and a modest arrest in both G1 and G2/M. Thus, in contrast to other studies, all tumour cells investigated showed either a typical p53wt or mutant (mut) pattern. Protein function was compared with cell survival and DNA damage, as assessed previously. p53 wild-type cells were on average 1.3-times (n.s.) more radiosensitive than mutant cells, but there was a considerable overlap between both groups. Further, the 1.3-fold enhanced resistance of cells lacking wild-type p53 was paralleled by a 1.3-fold lower number of induced double-strand breaks. The results suggest that p53 could have impact on chromatin compaction and thus effect DNA damage induction and radiosensitivity of tumour cells. PMID- 14761851 TI - Radiation-induced variations in urothelial expression of intercellular adhesion molecule 1 (ICAM-1): association with changes in urinary bladder function. AB - PURPOSE: To assess the effect of single-dose irradiation on intercellular adhesion molecule 1 (ICAM-1) expression in the urothelium of mouse urinary bladder and to correlate ICAM-1 variations with fluctuations in storage capacity during the early and late radiation response. MATERIALS AND METHODS: Groups of female C3H mice were subjected to irradiation with either 20 or 0 Gy. The intensity of immunohistochemical ICAM-1 staining in the urothelium was assessed in a semiquantitative way applying an arbitrary score (0-5). Changes in bladder storage function were assessed by transurethral cystometry. RESULTS: For the early radiation response phase, a reduction in bladder capacity by >50%, i.e. a positive functional radiation response, was seen in 40% of the irradiated animals between days 0 and 15, and in 64% of animals during days 16-30. During the late response phase, 71% of the animals sacrificed after day 180 developed a positive functional response. Urothelial cells were found to express ICAM-1 constitutively. Irradiation resulted in an early rise in staining signal by day 2, with a maximum on day 4 and a return to control values on day 13. A permanent increase in ICAM-1 staining signal was observed in the late phase, from day 90 to 360 after irradiation. The expression of ICAM-1 in animals with a positive late response was 4.2+/-1.2 (mean+/-standard deviation), compared with 2.6+/-1.0 in non-responders (p=0.0009). CONCLUSION: Irradiation induces significant acute and chronic changes in urothelial ICAM-1 expression indicating that the urothelium contributes to the pathogenesis of both acute and late radiation effects in the urinary bladder. PMID- 14761852 TI - Feasibility and limits of bone marrow mononuclear cell expansion following irradiation. AB - PURPOSE: To define the ability of bone marrow mononuclear cells (BMMNC) to expand after irradiation and to determine the amount of apoptosis in irradiated expanded cells. MATERIALS AND METHODS: Non-human primate BMMNC were irradiated in vitro at doses ranging from 0 to 4 Gy and were cultured during 1 week in the presence of interleukin 3, interleukin 6, stem cell factor, thrombopoietin and fms-like tyrosine kinase-3 ligand. The expansion yield of BMMNC, colony-forming cells and CD34(+) cells were compared with non-irradiated control cultures. Apoptosis in expanded cells was also defined by annexin V/propidium iodine staining. RESULTS: Irradiation of BMMNC up to 1 Gy did not modify the ability of haematopoietic cells to expand. At higher doses, expansion of haematopoietic cells is reduced as compared with non-irradiated cultures but it remains significant. This reduction in expansion of BMMNC was related to radiation-induced apoptosis. CONCLUSION: The results suggest that it is possible to expand haematopoietic cells after irradiation doses at least up to 2 Gy. This suggests a possible use of cell therapy for the treatment of radiation accident victims. PMID- 14761853 TI - Workshop on networking radiation sciences in health, safety and the environment. PMID- 14761860 TI - Identification of a novel region in the proximal promoter of the mouse renin gene critical for expression. AB - An enhancer at -2.6 kb and a HOX.PBX-binding site at -60 bp have been demonstrated to be critical to expression of the mouse renin gene (Ren-1(c)) in As4.1 cells. In this report, we show that a region (-197 to -70) immediately 5' to the HOX.PBX-binding site is also critical for Ren-1(c) expression. Deletion of this region in a construct containing 4.1 kb of the Ren-1(c) 5'-flanking sequence resulted in a 99% reduction in Ren-1(c) promoter activity in As4.1 cells, suggesting the pivotal role for the region in the regulation of the mouse renin gene. Electrophoretic mobility shift and supershift assays have identified two nuclear factor I-binding sites and a Sp1/Sp3-binding site within the distal portion of the region (-197 to -103). Mutation of these three sites caused a 90% decrease in Ren-1(c) promoter activity. Mutational analysis and electrophoretic mobility shift assays have also identified three additional transcription factor binding sites within the region from -103 to -69, each of which contributes to high-level expression of Ren-1(c) in As4.1 cells. Finally, we have shown that the Ren-1(c) enhancer is the target for endothelin-1 (ET-1)-induced inhibition of Ren 1(c) expression and the transcription factor-binding sites in the proximal promoter are required for the maximal ET-1 inhibitory effect. PMID- 14761861 TI - Effect of 2'-phosphophloretin on renal function in chronic renal failure rats. AB - Hyperhosphatemia and secondary hyperparathyroidism are common and severe complications of chronic renal failure. Therapies to reduce serum phosphate have been shown to reduce serum parathyroid hormone (PTH) and slow the progression of renal failure. The effect of the inhibitor of intestinal phosphate absorption, 2' phosphophloretin (2'-PP), on serum and urine chemistry, renal histology, and cardiac structure in the uremic rat model of renal failure, 5/6 nephrectomy (5/6 NX), was examined. The effect of 2'-PP on serum phosphate, serum PTH, serum total Ca(2+), and ionized Ca(2+), Ca(2+) x P(i) product, urine protein, urine osmolality, and creatinine clearance in 5/6 NX rats was examined. Uremic rats in chronic renal failure were gavaged daily with 25 microM 2'-PP. Over the course of a 5-wk experiment, serum chemistry in untreated uremic rats, 2'-PP-treated uremic rats, and age-matched control rats with normal renal function was determined twice a week. Urine creatinine, urine osmolality, urine phosphate, and urine protein were determined once a week from 24-h collections. 2'-PP reduced serum phosphate 40 +/- 3% compared with a 17% increase in untreated uremic control rats. 2'-PP did not alter total serum Ca(2+). During 5-wk experiments, serum PTH increased 65 +/- 25% in untreated uremic rats and decreased 70 +/- 7% in uremic rats treated with 25 microM 2'-PP. Creatinine clearance decreased 20% in untreated uremic rats compared with a 100% increase in 2'-PP-treated uremic rats. Urine protein decreased and urine osmolality increased in uremic rats treated with 2'-PP. The mechanism of the effect of 2'-PP on serum phosphate was inhibition of intestinal phosphate absorption. 2-PP inhibited intestinal phosphate absorption 50% without altering dietary protein absorption or intestinal Ca(2+) absorption. Over the course of the 5-wk treatment with 2'-PP, uremic animals treated with 2'-PP had a 2-4% weight gain/wk, similar to the weight gain seen in age-matched control rats with normal renal function. PMID- 14761862 TI - Chronic hyperaldosteronism in a transgenic mouse model fails to induce cardiac remodeling and fibrosis under a normal-salt diet. AB - Primary aldosteronism causes severe hypertension in humans (Conn's syndrome) with cardiac hypertrophy, characterized by a fibrosis more severe than the one observed in patients with essential hypertension. This suggests that aldosterone by itself may have specific and direct effects on cardiac remodeling through the activation of the cardiac mineralocorticoid receptor. Experimental evidence obtained in studying uninephrectomized rats treated with aldosterone or deoxycorticosterone (DOC) together with salt loading has led to similar conclusions. To examine the direct consequences of chronically elevated aldosterone levels on cardiac pathophysiology, we analyzed a mouse model (alpha epithelial Na channel -/-Tg) that is normotensive under normal-salt diet but exhibits chronic hyperaldosteronism. Sixteen-month-old transgenic rescue mice that were kept under a regular salt diet that contains a small amount of sodium (0.3% Na(+)) displayed a compensated PHA-1 phenotype with normal body weight, normal kidney index, normal blood pressure, but 6.3-fold elevated plasma aldosterone levels compared with the age-matched control group. Peripheral resistance of distal colon to aldosterone was shown by a significant decrease of the amiloride-sensitive rectal potential difference, and its diurnal cyclicity was blunted. Despite chronically high plasma aldosterone levels, these animals do not show any evidence of cardiac hypertrophy, remodeling, or fibrosis, using collagen staining and anti-alpha-skeletal and alpha-smooth actin immunochemical labeling of heart sections. Cardiac fibrosis as seen in DOC- or aldosterone/salt treated animal models is therefore likely to be due to the synergistic effect of salt, aldosterone, and other confounding factors rather than to the elevated circulating aldosterone levels alone. PMID- 14761863 TI - Gamma-MSH, sodium metabolism, and salt-sensitive hypertension. AB - Alpha-, beta-, and gamma-melanocyte stimulating hormones (MSHs) are melanotropin peptides that are derived from the ACTH/beta-endorphin prohormone proopiomelanocortin (POMC). They have been highly conserved through evolutionary development, although their functions in mammals have remained obscure. The identification in the last decade of a family of five membrane-spanning melanocortin receptors (MC-Rs), for which the melanotropins are the natural ligands, has permitted the characterization of a number of important actions of these peptides, although the physiological function(s) of gamma-MSH have remained elusive. Much evidence indicates that gamma-MSH stimulates sympathetic outflow and raises blood pressure through a central mechanism. However, this review focuses on newer cardiovascular and renal actions of the peptide, acting in most cases through the MC3-R. In rodents, a high-sodium diet (HSD) increases the pituitary abundance of POMC mRNA and of gamma-MSH content and results in a doubling of plasma gamma-MSH concentration. The peptide is natriuretic and acts through renal MC3-Rs, which are also upregulated by the HSD. Thus the system appears designed to participate in the integrated response to dietary sodium excess. Genetic or pharmacologic induction of gamma-MSH deficiency results in marked salt-sensitive hypertension that is corrected by the administration of the peptide, probably through a central site of action. Deletion of the MC3-R also produces salt-sensitive hypertension, which, however, is not corrected by infusion of the hormone. These observations in aggregate suggest the operation of a hormonal system important in blood pressure control and in the regulation of sodium excretion. The relationship of these two actions to each other and the significance of this system in humans are important questions for future research. PMID- 14761864 TI - Methods for detection of reactive metabolites of oxygen and nitrogen: in vitro and in vivo considerations. AB - Facile detection of reactive oxygen and nitrogen species in biologic systems is often problematic. This is a result of the numerous cellular mechanisms, both enzymatic and nonenzymatic involved in their catabolism/decomposition, the complex and overlapping nature of their reactivities, as well as the often limited intracellular access of detector systems. This review describes approaches to the direct and indirect measurement of different reactive metabolites of oxygen and nitrogen. Particular attention to a method's applicability for in vivo determinations will be addressed. PMID- 14761865 TI - Estrogen receptors and central osmotic regulation. PMID- 14761866 TI - Medullary pathways regulating sympathetic outflow: the need for more lateral thinking. PMID- 14761868 TI - Short-term fluoxetine treatment enhances baroreflex control of sympathetic nervous system activity after hindlimb unloading. AB - Data in humans indicate that individuals with orthostatic hypotension that are refractory to other traditional forms of therapy are responsive to selective serotonin reuptake inhibitor (SSRI) treatment. We tested the hypothesis that SSRI administration would help correct the attenuated baroreflex control of sympathetic nervous system activity in the hindlimb-unloaded (HU) rat model of cardiovascular deconditioning. An initial study was conducted to determine the time course of effects of fluoxetine (Flu) administration on baroreflex control of lumbar sympathetic nerve activity (LSNA) in conscious, chronically instrumented rats. Animals received either vehicle (Veh, sterile water) or 10 mg/kg Flu for 1, 4, or 16 days of treatment. Data indicate that while 1-day and 16-day Flu administration did not affect baroreflex function, baroreflex control of LSNA was enhanced after 4-day (short term) Flu administration. HU rats were then treated with Flu for 4 days and compared with HU rats receiving Veh and to casted control rats maintained in the normal posture that received either Veh or short-term Flu treatment. Similar to pilot data, short-term Flu treatment enhanced baroreflex control of LSNA in both HU rats and control rats. These data taken together indicate that baroreflex control of sympathetic nervous system activity is a possible mechanism responsible for the successful treatment of orthostatic intolerance with Flu. PMID- 14761867 TI - Dissecting the JGA: new functions for JG cells? PMID- 14761869 TI - Leukocyte inflammatory mediators and lung pathophysiology: an update. PMID- 14761870 TI - Weird and weirder: how circulating chemokines coax neutrophils to the lung. PMID- 14761871 TI - p21Waf1/Cip1 and the prevention of oxidative stress. PMID- 14761872 TI - Time and pressure dependence of transvascular Clara cell protein, albumin, and IgG transport during ventilator-induced lung injury in mice. AB - We compared the transport of three proteins with different hydrodynamic radii with ultrastructural changes in lungs of intact mice ventilated at peak inflation pressures (PIP) of 15, 35, 45, and 55 cmH(2)O for 2 h and PIP of 55 cmH(2)O for 0.5 and 1 h. After 2 h of ventilation, significant increases were observed in plasma Clara cell secretory protein (1.9 nm radius) at 35 cmH(2)O PIP and in bronchoalveolar lavage fluid albumin (3.6 nm radius) at 45 cmH(2)O PIP and IgG (5.6 nm radius) at 55 cmH(2)O PIP. Increased concentrations of all three proteins and lung wet-to-dry weight ratios were significantly correlated with PIP and ventilation time. Clara cell secretory protein and albumin increased significantly after 0.5 h of 55 cmH(2)O PIP, but IgG increased only after 2 h. Separation of endothelium or epithelium to form blebs was apparent only in small vessels (15-30 microm diameter) at 45 cmH(2)O PIP and after 0.5 h at 55 cmH(2)O PIP but became extensive after 2 h of ventilation at 55 cmH(2)O PIP. Junctional gaps between cells were rarely observed. Ultrastructural lung injury and protein clearances across the air-blood barrier were related to ventilation time and PIP levels. Protein clearances increased in relation to molecular size, consistent with increasing dimensions and frequency of transmembrane aqueous pathways. PMID- 14761873 TI - In vivo regulation of growth hormone-stimulated gene transcription by STAT5b. AB - The long-term effects of growth hormone (GH) are mediated through coordinated changes in gene expression that are the outcome of interactions between hormone activated signal transduction pathways and specific feedback loops. Recent studies in mice have implicated the transcription factor STAT5b as part of the GH regulated somatic growth pathway, because mice lacking this protein showed diminished growth rates. To assess the role of Stat5b in GH-stimulated gene expression, we have delivered modified versions of the protein to the liver of pituitary-deficient male rats by quantitative adenovirus-mediated gene transfer. In pilot studies in cell culture, both constitutive-active and dominant-negative STAT5b showed appropriate binding properties toward a specific DNA response element. After in vivo expression, neither protein prevented nuclear accumulation of STATs 1 and 3 in the liver. Dominant-negative STAT5b completely inhibited GH stimulated transcription of genes encoding the growth-promoting proteins IGF-I, IGF-binding protein-3 (IGFBP-3), and acid-labile subunit (ALS), which comprise the major circulating IGF-I complex, and blocked expression of the GH inhibitors SOCS-1, SOCS-2, and CIS, but had little effect on induction of SOCS-3. Constitutive-active STAT5b stimulated robust transcription of IGF-I, ALS, and IGFBP-3 in the absence of hormone but did little to modify GH-mediated activation of SOCS family genes. An adenovirus encoding EGFP was without effect. These results, in addition to establishing STAT5b as one of the key agents of GH stimulated gene transcription, demonstrate the feasibility of using in vivo gene transfer to target and dissect the functions of distinct components of complex hormone-activated signal transduction pathways. PMID- 14761874 TI - cAMP-positive regulation of angiotensinogen gene expression and protein secretion in rat adipose tissue. AB - The adipose renin-angiotensin system (RAS) has been assigned to participate in the control of adipose tissue development and in the pathogenesis of obesity related hypertension. In adipose cells, the biological responses to beta adrenergic stimulation are mediated by an increase in intracellular cAMP. Because cAMP is known to promote adipogenesis and because an association exists between body fat mass, hypertension, and increased sympathetic stimulation, we examined the influence of cAMP on angiotensinogen (ATG) expression and secretion in rat adipose tissue. Exposure of primary cultured differentiated preadipocytes to the cAMP analog 8-bromoadenosine 3',5'-cyclic monophosphate (8-BrcAMP) or cAMP stimulating agents (forskolin and IBMX) results in a significant increase in ATG mRNA levels. In adipose tissue fragments, 8-BrcAMP also increases ATG mRNA levels and protein secretion, but not in the presence of the protein kinase A inhibitor H89. The addition of isoproterenol, known to stimulate the synthesis of intracellular cAMP via beta-adrenoreceptors, had the same stimulatory effect on ATG expression and secretion. These results indicate that cAMP in vitro upregulates ATG expression and secretion in rat adipose tissue via the protein kinase A-dependent pathway. Further studies are required to determine whether this regulatory pathway is activated in human obesity, where increased sympathetic tone is frequently observed, and to elucidate the importance of adipose ATG to the elevated blood pressure observed in this pathological state. PMID- 14761875 TI - Postnatal ontogeny of glucose homeostasis and insulin action in sheep. AB - Glucose tolerance declines with maturation and aging in several species, but the time of onset and extent of changes in insulin sensitivity and insulin secretion and their contribution to changes in glucose tolerance are unclear. We therefore determined the effect of maturation on glucose tolerance, insulin secretion, and insulin sensitivity in a longitudinal study of male and female sheep from preweaning to adulthood, and whether these measures were related across age. Glucose tolerance was assessed by intravenous glucose tolerance test (IVGTT, 0.25 g glucose/kg), insulin secretion as the integrated insulin concentration during IVGTT, and insulin sensitivity by hyperinsulinemic-euglycemic clamp (2 mU insulin.kg(-1).min(-1)). Glucose tolerance, relative insulin secretion, and insulin sensitivity each decreased with age (P < 0.001). The disposition index, the product of insulin sensitivity, and various measures of insulin secretion during fasting or IVGTT also decreased with age (P < 0.001). Glucose tolerance in young adult sheep was independently predicted by insulin sensitivity (P = 0.012) and by insulin secretion relative to integrated glucose during IVGTT (P = 0.005). Relative insulin secretion before weaning was correlated positively with that in the adult (P = 0.023), whereas glucose tolerance, insulin sensitivity, and disposition indexes in the adult did not correlate with those at earlier ages. We conclude that glucose tolerance declines between the first month of life and early adulthood in the sheep, reflecting decreasing insulin sensitivity and absence of compensatory insulin secretion. Nevertheless, the capacity for insulin secretion in the adult reflects that early in life, suggesting that it is determined genetically or by persistent influences of the perinatal environment. PMID- 14761876 TI - Regulation of neonatal liver protein synthesis by insulin and amino acids in pigs. AB - The high efficiency of protein deposition during the neonatal period is driven by high rates of protein synthesis, which are maximally stimulated after feeding. Infusion of amino acids, but not insulin, reproduces the feeding-induced stimulation of liver protein synthesis. To determine whether amino acid stimulated liver protein synthesis is independent of insulin in neonates, and to examine the role of amino acids and insulin in the regulation of translation initiation in neonatal liver, we performed pancreatic glucose-amino acid clamps in overnight-fasted 7-day-old pigs. Pigs (n = 9-12/group) were infused with insulin at 0, 10, 22, and 110 ng.kg(-0.66).min(-1) to achieve 0, 2, 6, and 30 microU/ml insulin, respectively. At each insulin dose, amino acids were maintained at fasting or fed levels or, in conjunction with the highest insulin dose, allowed to fall to below fasting levels. Insulin had no effect on the fractional rate of protein synthesis in liver. Amino acids increased fractional protein synthesis rates in liver at each dose of insulin, including the 0 microU/ml dose. There was a dose-response effect of amino acids on liver protein synthesis. Amino acids and insulin increased protein S6 kinase and 4E-binding protein 1 (4E-BP1) phosphorylation; however, only amino acids decreased formation of the inactive 4E-BPI.eukaryotic initiation factor-4E (eIF4E) complex. The results suggest that amino acids regulate liver protein synthesis in the neonate by modulating the availability of eIF4E for 48S ribosomal complex formation and that this response does not require insulin. PMID- 14761877 TI - Regulation of prostate-specific antigen by activin A in prostate cancer LNCaP cells. AB - Activins are multifunctional growth and differentiation factors and stimulate FSH beta gene expression and FSH secretion by the pituitary gonadotropes. Follistatins bind activin, resulting in the neutralization of activin bioactivity. The activin/follistatin system is present in the prostate tissue. Prostate-specific antigen (PSA) plays an important role in male reproductive physiology as well as being very important as a tumor marker for prostate cancer. Thus the regulation of PSA has important clinical implications. Previous studies showed that PSA is primarily regulated by androgens. In the present study, we evaluated the direct effects of activin A on the proliferation and PSA production of prostate cancer LNCaP cells, which express functional activin receptors and androgen receptor and PSA. LNCaP cells were treated with activin A and 5alpha dihydrotestosterone (DHT) with or without their antagonists (follistatin or the nonsteroidal anti-androgen bicalutamide). Activin A decreased cell growth of LNCaP cells in a dose-dependent manner, whereas DHT increased it in a biphasic manner. In contrast to their opposing actions on cell growth, both activin A and DHT upregulated PSA gene expression and increased PSA secretion by LNCaP cells. The effects of activin A and DHT to increase PSA production were synergistic or additive. Follistatin or bicalutamide was without effect on cell growth or PSA production. The effects of activin A on LNCaP cells were blocked by follistatin, not by bicalutamide, whereas effects of DHT were prevented by bicalutamide, not by follistatin. Activin A upregulates PSA production, and the effect is through an androgen receptor-independent pathway. The activin/follistatin system can be a physiological modulator of PSA gene transcription and secretion in the prostate tissue, and activins may cooperate with androgen to upregulate PSA in vivo. PMID- 14761878 TI - Regulation of metabolic genes in human skeletal muscle by short-term exercise and diet manipulation. AB - Changes in dietary macronutrient intake alter muscle and blood substrate availability and are important for regulating gene expression. However, few studies have examined the effects of diet manipulation on gene expression in human skeletal muscle. The aim of this study was to quantify the extent to which altering substrate availability impacts on subsequent mRNA abundance of a subset of carbohydrate (CHO)- and fat-related genes. Seven subjects consumed either a low- (LOW; 0.7 g/kg body mass CHO) or high- (HIGH; 10 g/kg body mass CHO) CHO diet for 48 h after performing an exhaustive exercise bout to deplete muscle glycogen stores. After intervention, resting muscle and blood samples were taken. Muscle was analyzed for the gene abundances of GLUT4, glycogenin, pyruvate dehydrogenase kinase-4 (PDK-4), fatty acid translocase (FAT/CD36), carnitine palmitoyltransferase I (CPT I), hormone-sensitive lipase (HSL), beta-hydroxyacyl CoA dehydrogenase (beta-HAD), and uncoupling binding protein-3 (UCP3), and blood samples for glucose, insulin, and free fatty acid (FFA) concentrations. Glycogen depleting exercise and HIGH-CHO resulted in a 300% increase in muscle glycogen content (P < 0.001) relative to the LOW-CHO condition. FFA concentrations were twofold higher after LOW- vs. HIGH-CHO (P < 0.05). The exercise-diet manipulation exerted a significant effect on transcription of all carbohydrate-related genes, with an increase in GLUT4 and glycogenin mRNA abundance and a reduction in PDK-4 transcription after HIGH-CHO (all P < 0.05). FAT/CD36 (P < 0.05) and UCP3 (P < 0.01) gene transcriptions were increased following LOW-CHO. We conclude that 1) there was a rapid capacity for a short-term exercise and diet intervention to exert coordinated changes in the mRNA transcription of metabolic related genes, and 2) genes involved in glucose regulation are increased following a high carbohydrate diet. PMID- 14761879 TI - Drug discovery in academia. AB - Drug discovery and development is generally done in the commercial rather than the academic realm. Drug discovery involves target discovery and validation, lead identification by high-throughput screening, and lead optimization by medicinal chemistry. Follow-up preclinical evaluation includes analysis in animal models of compound efficacy and pharmacology (ADME: administration, distribution, metabolism, elimination) and studies of toxicology, specificity, and drug interactions. Notwithstanding the high-cost, labor-intensive, and non-hypothesis driven aspects of drug discovery, the academic setting has a unique and expanding niche in this important area of investigation. For example, academic drug discovery can focus on targets of limited commercial value, such as third-world and rare diseases, and on the development of research reagents such as high affinity inhibitors for pharmacological "gene knockout" in animal models ("chemical genetics"). This review describes the practical aspects of the preclinical drug discovery process for academic investigators. The discovery of small molecule inhibitors and activators of the cystic fibrosis transmembrane conductance regulator is presented as an example of an academic drug discovery program that has yielded new compounds for physiology research and clinical development. PMID- 14761880 TI - Has HGF met other partners? Met-independent epithelial morphogenesis induced by HGF. focus on "Hepatocyte growth factor induces MDCK cell morphogenesis without causing loss of tight junction functional integrity". PMID- 14761881 TI - K+ channels in the secretory membrane of the parietal cell. focus on "Gastric parietal cell secretory membrane contains PKA- and acid-activated Kir2.1 K+ channels". PMID- 14761882 TI - Rapamycin plays a new role as differentiator of vascular smooth muscle phenotype. focus on "The mTOR/p70 S6K1 pathway regulates vascular smooth muscle differentiation". PMID- 14761883 TI - Extracellular matrix controls myosin light chain phosphorylation and cell contractility through modulation of cell shape and cytoskeletal prestress. AB - The mechanism by which vascular smooth muscle (VSM) cells modulate their contractility in response to structural cues from extracellular matrix remains poorly understood. When pulmonary VSM cells were cultured on increasing densities of immobilized fibronectin (FN), cell spreading, myosin light chain (MLC) phosphorylation, cytoskeletal prestress (isometric tension in the cell before vasoagonist stimulation), and the active contractile response to the vasoconstrictor endothelin-1 all increased in parallel. In contrast, MLC phosphorylation did not increase when suspended cells were allowed to bind FN coated microbeads (4.5-microm diameter) or cultured on micrometer-sized (30 x 30 microm) FN islands surrounded by nonadhesive regions that support integrin binding but prevent cell spreading. Cell spreading and MLC phosphorylation also both decreased in parallel when the mechanical compliance of flexible FN substrates was raised. MLC phosphorylation was inhibited independently of cell shape when cytoskeletal prestress was dissipated using a myosin ATPase inhibitor in fully spread cells, whereas it increased to maximal levels when microtubules were disrupted using nocodazole in cells adherent to FN but not in suspended cells. These data demonstrate that changes in cell-extracellular matrix (ECM) interactions modulate smooth muscle cell contractility at the level of biochemical signal transduction and suggest that the mechanism underlying this regulation may involve physical interplay between ECM and the cytoskeleton, such that cell spreading and generation of cytoskeletal tension feed back to promote MLC phosphorylation and further increase tension generation. PMID- 14761885 TI - Control of Na+-K+-ATPase beta 1-subunit expression: role of 3'-untranslated region. AB - Using in vitro translation and cell transfection assays, we previously demonstrated that the Na+ -K+ -ATPase beta1 mRNA species containing its longest 3'-untranslated region (UTR) exhibited the lowest translational efficiency. Here, employing deletions and in vivo expression assays, using direct injection of plasmids into rat ventricular myocardium, we identified a 143-nt segment located in the distal 3'-UTR of beta1 mRNA that was associated with decreased luciferase expression; interestingly, this segment contains three AUUUA motifs. Using RNA protein binding assays and UV cross-linking of cRNA with cytosolic proteins of rat heart, we identified an approximately 38-kDa protein that specifically bound to the cRNA encoding the 143-nt segment of beta1 mRNA 3'-UTR. Mutation of three nucleotides located in the middle region of the 143-nt segment, which was predicted to greatly disrupt a putative stem-loop structure of the cRNA in this region, was associated with reduced binding of the mutated cRNA to the protein migrating at approximately 38 kDa. The cRNA encoding a segment of cyclooxygenase 2 mRNA 3'-UTR containing six AUUUA sequences did not bind the protein migrating at approximately 38 kDa and did not compete with the binding of the wild-type 143 nt beta(1) cRNA to the protein. The above results suggest that the 143-nt segment in the distal segment of the 3'-UTR of beta1 mRNA may play an important role in the control of beta1-subunit expression. PMID- 14761884 TI - Tumor necrosis factor activates CRE-binding protein through a p38 MAPK/MSK1 signaling pathway in endothelial cells. AB - Tumor necrosis factor (TNF) promotes immunity and modulates cell viability, in part, by promoting alterations of cellular gene expression. The mechanisms through which TNF communicates with the nucleus and alters gene expression are incompletely understood. Incubation of human umbilical vein endothelial cells (HUVEC) with TNF induces phosphorylation of the CRE-binding protein (CREB) transcription factor on serine 133 and increases CREB DNA binding and transactivation. Dominant negative CREB, an antagonist antibody directed against the type 1 TNF receptor, or pharmacological inhibition of p38 MAPK signaling blocked TNF-induced CREB activation as determined by phosphorylation and gene reporter assays. From among the kinases that can activate CREB, we found that downstream of p38 MAPK, MSK1 is activated by TNF to promote CREB activation. These observations show that CREB is activated by TNF/TNFR1 signaling through a p38MAPK/MSK1 signaling pathway. PMID- 14761886 TI - Characterization of G proteins involved in activation of nonselective cation channels and arachidonic acid release by norepinephrine/alpha1A-adrenergic receptors. AB - We demonstrated recently that norepinephrine activates Ca2+ -permeable nonselective cation channels (NSCCs) in Chinese hamster ovary cells stably expressing alpha1A-adrenergic receptors (CHO-alpha1A). Moreover, extracellular Ca2+ through NSCCs plays essential roles in norepinephrine-induced arachidonic acid release. The purpose of the present study was to identify the G proteins involved in the activation of NSCCs and arachidonic acid release by norepinephrine. For these purposes, we used U73122, an inhibitor of phospholipase C (PLC), and dominant negative mutants of G12 and G13 (G12G228A and G13G225A, respectively). U73122 failed to inhibit NSCCs activation by norepinephrine. The magnitudes of norepinephrine-induced extracellular Ca2+ influx in CHO-alpha1A microinjected with G13G225A were smaller than those in CHO-alpha1A. In contrast, the magnitudes of norepinephrine-induced extracellular Ca2+ influx in CHO-alpha1A microinjected with G12G228A were similar to those in CHO-alpha1A. In addition, neither a Rho-associated kinase (ROCK) inhibitor nor a phosphoinositide 3-kinase inhibitor affected norepinephrine-induced extracellular Ca2+ influx. G13G225A, but not G12G228A, also inhibited arachidonic acid release partially. These results demonstrate that 1) the Gq/PLC-pathway is not involved in NSCCs activation by norepinephrine, 2) G13 couples with CHO-alpha1A and plays important roles for norepinephrine-induced NSCCs activation, 3) neither ROCK- nor PI3K dependent cascade is involved in NSCCs activation, and 4) G13 is involved in norepinephrine-induced arachidonic acid release in CHO-alpha1A. PMID- 14761887 TI - Evidence for functional estrogen receptors alpha and beta in human adipose cells: regional specificities and regulation by estrogens. AB - Adipocytes are estrogen-responsive cells, but the quantitative expression and transcriptional regulation of the estrogen receptors (ER-alpha and ER-beta) in human adipocytes and their precursor cells are unclear. Using real-time quantitative PCR, we have demonstrated that both ER-alpha and ER-beta mRNA are expressed in human mature adipocytes with a large predominance of ER-alpha mRNA. Moreover, ER-alpha mRNA is identically expressed whatever the anatomic origin (intraabdominal and subcutaneous) of the adipocytes and the gender. ER-beta mRNA levels are higher in women compared with men, without regional differences. 17beta-Estradiol in vitro upregulates expression of both ER-alpha and ER-beta mRNA in subcutaneous adipocytes from women but only the ER-alpha mRNA in subcutaneous and intra-abdominal adipocytes from men. In preadipocytes, only the ER-alpha subtype was present. In the latter cells, estrogens in vitro had no influence on ER-alpha expression (mRNA and protein). The present study also shows that estrogens in vitro increase the AP-1, SP-1, and estrogen response element DNA binding activities in differentiated but not in confluent preadipocytes, suggesting that ER become functional during the course of adipogenesis. On the whole, these data are consistent with a predominant role of the ER-alpha subtype in mediating the effects of estrogens on human adipose tissue development and metabolism. PMID- 14761888 TI - M-cadherin transcription in satellite cells from normal and denervated muscle. AB - Satellite cells (SC) in adult muscle are quiescent in the G0 phase of the cell cycle. In the present study we determined whether SC after denervation upregulate M-cadherin, an adhesion molecule that is upregulated with differentiation and fusion. We also monitored primary cultures of SC from denervated muscle for expression of the transcription factors of the MyoD family to determine whether SC from denervated muscle can be activated in vitro. Hindlimb muscles of rats were denervated under anesthesia, and rats were killed after 2-28 days. The SC of the denervated limbs were pooled and either assessed for M-cadherin mRNA by using real-time RT-PCR or cultured in vitro. The cultures were processed for RT-PCR or immunofluorescence for expression of the transcription factors of the MyoD family. Hindlimb muscles of M-cadherin knockout mice were denervated under anesthesia, mice were killed after 2-28 days, and cells were stained for beta galactosidase activity by X-gal histochemistry. In vitro, primary SC cultures from rat muscle denervated for 2-28 days expressed transcripts of myf5, MyoD, myogenin, and MRF4 as SC from normal innervated muscle. In vivo, M-cadherin transcription was not upregulated in SC from denervated rat muscle when compared with normal muscle. Moreover, beta-galactosidase activity was not detected in denervated mouse muscle. The finding that SC do not upregulate M-cadherin after denervation supports the notion that they remain in the G(0) phase of the cell cycle in vivo. However, the cells retain the capacity to pass through the proliferative and differentiative program when robustly stimulated to do so in vitro. PMID- 14761889 TI - Differential membrane potential and ion current responses to different types of shear stress in vascular endothelial cells. AB - Vascular endothelial cells (ECs) distinguish among and respond differently to different types of fluid mechanical shear stress. Elucidating the mechanisms governing this differential responsiveness is the key to understanding why early atherosclerotic lesions localize preferentially in arterial regions exposed to low and/or oscillatory flow. An early and very rapid endothelial response to flow is the activation of flow-sensitive K(+) and Cl(-) channels that respectively hyperpolarize and depolarize the cell membrane and regulate several important endothelial responses to flow. We have used whole cell current- and voltage-clamp techniques to demonstrate that flow-sensitive hyperpolarizing and depolarizing currents respond differently to different types of shear stress in cultured bovine aortic ECs. A steady shear stress level of 10 dyn/cm(2) activated both currents leading to rapid membrane hyperpolarization that was subsequently reversed to depolarization. In contrast, a steady shear stress of 1 dyn/cm(2) only activated the hyperpolarizing current. A purely oscillatory shear stress of 0 +/- 10 dyn/cm(2) with an oscillation frequency of either 1 or 0.2 Hz activated the hyperpolarizing current but only minimally the depolarizing current, whereas a 5-Hz oscillation activated neither current. These results demonstrate for the first time that flow-activated ion currents exhibit different sensitivities to shear stress magnitude and oscillation frequency. We propose that flow-sensitive ion channels constitute components of an integrated mechanosensing system that, through the aggregate effect of ion channel activation on cell membrane potential, enables ECs to distinguish among different types of flow. PMID- 14761890 TI - Electron microprobe analysis of ouabain-exposed ciliary epithelium: PE-NPE cell couplets form the functional units. AB - Aqueous humor is secreted by the bilayered ciliary epithelium. Solutes and water enter the pigmented ciliary epithelial (PE) cell layer, cross gap junctions into the nonpigmented ciliary epithelial (NPE) cell layer, and are released into the aqueous humor. Electrical measurements suggest that heptanol reduces transepithelial ion movement by interrupting PE-NPE communication and that gap junctions may be a regulatory site of aqueous humor formation. Several lines of evidence also suggest that net ciliary epithelial transport is strongly region dependent. Divided rabbit iris-ciliary bodies were incubated in chambers under control and experimental conditions, quick-frozen, cryosectioned, and freeze dried. Elemental intracellular contents of NPE and PE cells were determined by electron probe X-ray microanalysis. With or without heptanol, ouabain produced concentration- and time-dependent changes more markedly in anterior than in posterior epithelium. Without heptanol, there were considerable cell-to-cell variations in Na gain and K loss. However, contiguous NPE and PE cells displayed similar changes, even when nearby cell pairs were little changed by ouabain in aqueous, stromal, or both reservoirs. In contrast, with heptanol present, ouabain added to aqueous or both reservoirs produced much larger changes in NPE than in PE cells. The results indicate that 1) heptanol indeed interrupts PE-NPE junctions, providing an opportunity for electron microprobe analysis of the sidedness of modification of ciliary epithelial secretion; 2) Na and K undergo faster turnover in anterior than in posterior epithelium; and 3) PE-NPE gap junctions differ from PE-PE and NPE-NPE junctions in permitting ionic equilibration between adjoining ouabain-stressed cells. PMID- 14761891 TI - An LQT mutant minK alters KvLQT1 trafficking. AB - Cardiac I(Ks), the slowly activated delayed-rectifier K(+) current, is produced by the protein complex composed of alpha- and beta-subunits: KvLQT1 and minK. Mutations of genes encoding KvLQT1 and minK are responsible for the hereditary long QT syndrome (loci LQT1 and LQT5, respectively). MinK-L51H fails to traffic to the cell surface, thereby failing to produce effective I(Ks). We examined the effects that minK-L51H and an endoplasmic reticulum (ER)-targeted minK (minK-ER) exerted over the electrophysiology and biosynthesis of coexpressed KvLQT1. Both minK-L51H and minK-ER were sequestered primarily in the ER as confirmed by lack of plasma membrane expression. Glycosylation and immunofluorescence patterns of minK-L51H were qualitatively different for minK-ER, suggesting differences in trafficking. Cotransfection with the minK mutants resulted in reduced surface expression of KvLQT1 as assayed by whole cell voltage clamp and immunofluorescence. MinK-L51H reduced current amplitude by 91% compared with wild type (WT) minK/KvLQT1, and the residual current was identical to KvLQT1 without minK. The phenotype of minK-L51H on I(Ks) was not dominant because coexpressed WT minK rescued the current and surface expression. Collectively, our data suggest that ER quality control prevents minK-L51H/KvLQT1 complexes from trafficking to the plasma membrane, resulting in decreased I(Ks). This is the first demonstration that a minK LQT mutation is capable of conferring trafficking defects onto its associated alpha-subunit. PMID- 14761892 TI - Collagen I upregulates extracellular matrix gene expression and secretion of TGF beta 1 by cultured human mesangial cells. AB - Progressive renal diseases are characterized by an increased synthesis of extracellular matrix (ECM) components. The mechanisms involved in the development of these alterations are not completely known, but a crucial role for TGF-beta 1 has been suggested. Moreover, the ability of the ECM to modulate the phenotypic expression of different cell types has been widely described. In experiments presented here, human mesangial cells (HMC) were grown on collagen type I (COL I) or IV (COL IV). ECM protein and TGF-beta 1 mRNA expression were evaluated by Northern blot analysis, and TGF-beta 1 secretion was evaluated by ELISA. The involvement of tyrosine kinase and serine-threonine kinase pathways was studied by Western blot analysis, immunofluorescence, and in vitro kinase assays. HMC cultured on COL I showed an increased mRNA expression of COL I and COL IV, fibronectin, and TGF-beta 1. Both tyrosine phosphorylation and integrin-linked kinase (ILK) activity increased when HMC were cultured on COL I, and blockade of these pathways inhibited the increased secretion of TGF-beta 1. In conclusion, the present results support a role for extracellular COL I in the regulation of TGF-beta 1 synthesis during progressive renal sclerosis and fibrosis and the subsequent increase in newly synthesized ECM proteins. In addition, ILK, along with the tyrosine kinases, participates in the genesis of this effect. PMID- 14761893 TI - Vacuolar H+-ATPase in human breast cancer cells with distinct metastatic potential: distribution and functional activity. AB - Tumor cells thrive in a hypoxic microenvironment with an acidic extracellular pH. To survive in this harsh environment, tumor cells must exhibit a dynamic cytosolic pH regulatory system. We hypothesize that vacuolar H(+)-ATPases (V ATPases) that normally reside in acidic organelles are also located at the cell surface, thus regulating cytosolic pH and exacerbating the migratory ability of metastatic cells. Immunocytochemical data revealed for the first time that V ATPase is located at the plasma membrane of human breast cancer cells: prominent in the highly metastatic and inconspicuous in the lowly metastatic cells. The V ATPase activities in isolated plasma membranes were greater in highly than in lowly metastatic cells. The proton fluxes via V-ATPase evaluated by fluorescence spectroscopy in living cells were greater in highly than in lowly metastatic cells. Interestingly, lowly metastatic cells preferentially used the ubiquitous Na(+)/H(+) exchanger and HCO(3)(-)-based H(+)-transporting mechanisms, whereas highly metastatic cells used plasma membrane V-ATPases. The highly metastatic cells were more invasive and migratory than the lowly metastatic cells. V-ATPase inhibitors decreased the invasion and migration in the highly metastatic cells. Altogether, these data indicate that V-ATPases located at the plasma membrane are involved in the acquisition of a more metastatic phenotype. PMID- 14761894 TI - Modulation of vascular smooth muscle cell migration by calcium/ calmodulin dependent protein kinase II-delta 2. AB - Previous studies demonstrated a requirement for multifunctional Ca(2+)/calmodulin dependent protein kinase II (CaMKII) in PDGF-stimulated vascular smooth muscle (VSM) cell migration. In the present study, molecular approaches were used specifically to assess the role of the predominant CaMKII isoform (delta(2) or delta(C)) on VSM cell migration. Kinase-negative (K43A) and constitutively active (T287D) mutant forms of CaMKII delta(2) were expressed using recombinant adenoviruses. CaMKII activities were evaluated in vitro by using a peptide substrate and in intact cells by assessing the phosphorylation of overexpressed phospholamban on Thr(17), a CaMKII-selective phosphorylation site. Expression of kinase-negative CaMKII delta(2) inhibited substrate phosphorylation both in vitro and in the intact cell, indicating a dominant-negative function with respect to exogenous substrate. However, overexpression of the kinase-negative mutant failed to inhibit endogenous CaMKII delta(2) autophosphorylation on Thr(287) after activation of cells with ionomycin, and in fact, these subunits served as a substrate for the endogenous kinase. Constitutively active CaMKII delta(2) phosphorylated substrate in vitro without added Ca(2+)/calmodulin and in the intact cell without added Ca(2+)-dependent stimuli, but it inhibited autophosphorylation of endogenous CaMKII delta(2) on Thr(287). Basal and PDGF stimulated cell migration was significantly enhanced in cells expressing kinase negative CaMKII delta(2), an effect opposite that of KN-93, a chemical inhibitor of CaMKII activation. Expression of the constitutively active CaMKII delta(2) mutant inhibited PDGF-stimulated cell migration. These studies point to a role for the CaMKII delta(2) isoform in regulating VSM cell migration. An inclusive interpretation of results using both pharmacological and molecular approaches raises the hypothesis that CaMKII delta(2) autophosphorylation may play an important role in PDGF-stimulated VSM cell migration. PMID- 14761896 TI - THE BIOFILM CONCEPT: CONSEQUENCES FOR FUTURE PROPHYLAXIS OF ORAL DISEASES? AB - Biofilm control is fundamental to oral health. Existing oral prophylactic measures, however, are insufficient. The main reason is probably because the micro-organisms involved organize into complex biofilm communities with features that differ from those of planktonic cells. Micro-organisms have traditionally been studied in the planktonic state. Conclusions drawn from many of these studies, therefore, need to be revalidated. Recent global approaches to the study of microbial gene expression and regulation in non-oral micro-organisms have shed light on two-component and quorum-sensing systems for the transduction of stimuli that allow for coordinated gene expression. We suggest interference with two component and quorum-sensing systems as potential novel strategies for the prevention of oral diseases through control of oral biofilms. Information is still lacking, however, on the genetic regulation of oral biofilm formation. A better understanding of these processes is of considerable importance. PMID- 14761895 TI - Extracellular pressure stimulates macrophage phagocytosis by inhibiting a pathway involving FAK and ERK. AB - We hypothesized that changes in extracellular pressure during inflammation or infection regulate macrophage phagocytosis through modulating the focal adhesion kinase (FAK)-ERK pathway. Undifferentiated (monocyte-like) or PMA-differentiated (macrophage-like) THP-1 cells were incubated at 37 degrees C with serum-opsonized latex beads under ambient or 20-mmHg increased pressure. Pressure did not affect monocyte phagocytosis but significantly increased macrophage phagocytosis (29.9 +/- 1.8 vs. 42.0 +/- 1.6%, n = 9, P < 0.001). THP-1 macrophages constitutively expressed activated FAK, ERK, and Src. Exposure of macrophages to pressure decreased ERK and FAK-Y397 phosphorylation (77.6 +/- 7.9%, n = 7, P < 0.05) but did not alter FAK-Y576 or Src phosphorylation. FAK small interfering RNA (SiRNA) reduced FAK expression by >75% and the basal amount of phosphorylated FAK by 25% and significantly increased basal macrophage phagocytosis (P < 0.05). Pressure inhibited FAK-Y397 phosphorylation in mock-transfected or scrambled SiRNA transfected macrophages, but phosphorylated FAK was not significantly reduced further by pressure in cells transfected with FAK SiRNA. Pressure increased phagocytosis in all three groups. However, FAK-SiRNA-transfected cells exhibited only 40% of the pressure effect on phagocytosis observed in scrambled SiRNA transfected cells so that phagocytosis inversely paralleled FAK activation. PD 98059 (50 microM), an ERK activation inhibitor, increased basal phagocytosis (26.9 +/- 1.8 vs. 31.7 +/- 1.1%, n = 15, P < 0.05), but pressure did not further increase phagocytosis in PD-98059-treated cells. Pressure also inhibited ERK activation after mock transfection or transfection with scrambled SiRNA, but transfection of FAK SiRNA abolished ERK inhibition by pressure. Pressure did not increase phagocytosis in MonoMac-1 cells that do not express FAK. Increased extracellular pressure during infection or inflammation enhances macrophage phagocytosis by inhibiting FAK and, consequently, decreasing ERK activation. PMID- 14761897 TI - CELLS AND EXTRACELLULAR MATRICES OF DENTIN AND PULP: A BIOLOGICAL BASIS FOR REPAIR AND TISSUE ENGINEERING. AB - Odontoblasts produce most of the extracellular matrix (ECM) components found in dentin and implicated in dentin mineralization. Major differences in the pulp ECM explain why pulp is normally a non-mineralized tissue. In vitro or in vivo, some dentin ECM molecules act as crystal nucleators and contribute to crystal growth, whereas others are mineralization inhibitors. After treatment of caries lesions of moderate progression, odontoblasts and cells from the sub-odontoblastic Hohl's layer are implicated in the formation of reactionary dentin. Healing of deeper lesions in contact with the pulp results in the formation of reparative dentin by pulp cells. The response to direct pulp-capping with materials such as calcium hydroxide is the formation of a dentinal bridge, resulting from the recruitment and proliferation of undifferentiated cells, which may be either stem cells or dedifferentiated and transdifferentiated mature cells. Once differentiated, the cells synthesize a matrix that undergoes mineralization. Animal models have been used to test the capacity of potentially bioactive molecules to promote pulp repair following their implantation into the pulp. ECM molecules induce either the formation of dentinal bridges or large areas of mineralization in the coronal pulp. They may also stimulate the total closure of the pulp in the root canal. In conclusion, some molecules found in dentin extracellular matrix may have potential in dental therapy as bioactive agents for pulp repair or tissue engineering. PMID- 14761898 TI - ORAL ADVERSE DRUG REACTIONS TO CARDIOVASCULAR DRUGS. AB - A great many cardiovascular drugs (CVDs) have the potential to induce adverse reactions in the mouth. The prevalence of such reactions is not known, however, since many are asymptomatic and therefore are believed to go unreported. As more drugs are marketed and the population includes an increasing number of elderly, the number of drug prescriptions is also expected to increase. Accordingly, it can be predicted that the occurrence of adverse drug reactions (ADRs), including the oral ones (ODRs), will continue to increase. ODRs affect the oral mucous membrane, saliva production, and taste. The pathogenesis of these reactions, especially the mucosal ones, is largely unknown and appears to involve complex interactions among the drug in question, other medications, the patient's underlying disease, genetics, and life-style factors. Along this line, there is a growing interest in the association between pharmacogenetic polymorphism and ADRs. Research focusing on polymorphism of the cytochrome P450 system (CYPs) has become increasingly important and has highlighted the intra- and inter-individual responses to drug exposure. This system has recently been suggested to be an underlying candidate regarding the pathogenesis of ADRs in the oral mucous membrane. This review focuses on those CVDs reported to induce ODRs. In addition, it will provide data on specific drugs or drug classes, and outline and discuss recent research on possible mechanisms linking ADRs to drug metabolism patterns. Abbreviations used will be as follows: ACEI, ACE inhibitor; ADR, adverse drug reaction; ANA, antinuclear antigen; ARB, angiotensin II receptor blocker; BAB, beta-adrenergic blocker; CCB, calcium-channel blocker; CDR, cutaneous drug reaction; CVD, cardiovascular drug; CYP, cytochrome P450 enzyme; EM, erythema multiforme; FDE, fixed drug eruption; I, inhibitor of CYP isoform activity; HMG CoA, hydroxymethyl-glutaryl coenzyme A; NAT, N-acetyltransferase; ODR, oral drug reaction; RDM, reactive drug metabolite; S, substrate for CYP isoform; SJS, Stevens-Johnson syndrome; SLE, systemic lupus erythematosus; and TEN, toxic epidermal necrolysis. PMID- 14761899 TI - BIOLOGICAL RISKS OF RESIN-BASED MATERIALS TO THE DENTIN-PULP COMPLEX. AB - Over the past 30 years, restorative dentistry has seen a revolution in materials, restorative techniques, and patient priorities. This revolution has been made possible with the development of new resin-based materials which can be bonded to the tooth structure. Not all of these changes have been without controversy or concern, and some have raised questions about the biological safety of these new materials and techniques. It is the purpose of this review to present recent and relevant information about the biological risks and consequences of resin-tooth bonding and how these risks are affected by the material, its clinical properties, and its manipulation by the practitioner. These biological risks are complex and interactive, and are still incompletely defined. In broad terms, these risks can be divided into those stemming from the toxicological properties of the materials themselves (direct biological risks) and those stemming from microbiological leakage (indirect biological risks). PMID- 14761901 TI - Tracking women's awareness of heart disease: an American Heart Association national study. AB - BACKGROUND: Cardiovascular disease (CVD) is the leading cause of mortality in men and women in the United States, yet prior research has shown a lack of awareness of risk among women. The purpose of this study was to assess the contemporary awareness, knowledge, and perceptions related to CVD risk among American women and to evaluate trends since 1997, when the American Heart Association initiated a national campaign to improve awareness of CVD among women. METHODS AND RESULTS: A telephone survey of a nationally representative random sample of women was conducted in June and July 2003, with an oversampling of black and Hispanic women; results were compared with those of similar surveys in 2000 and 1997. The present survey included 1024 respondents age > or =25 years; 68% were white, 12% black, 12% Hispanic, and 8% other ethnicities. Awareness, knowledge, and perceptions about heart disease were evaluated by use of a standard interviewer assisted questionnaire. A shift in awareness of heart disease as the leading killer of women has occurred since 1997. In 2003, 46% of respondents spontaneously identified heart disease as the leading cause of death in women, up from 30% in 1997 (P<0.05) and 34% in 2000 (P<0.05). In contrast, the percentage of women citing cancer as leading cause of death has significantly decreased. Black, Hispanic, and younger women (<45 years old) had lower awareness of heart disease as their leading cause of death than did white and older women. Nearly all women reported comfort in discussing prevention with healthcare providers, but only 38% of women reported that their doctors had ever discussed heart disease with them. CONCLUSIONS: Awareness of CVD has increased, although a significant gap between perceived and actual risk of CVD remains. Educational interventions to improve awareness and knowledge are needed, particularly for minority and younger women. PMID- 14761900 TI - Evidence-based guidelines for cardiovascular disease prevention in women. PMID- 14761902 TI - The role of human parietal cortex in attention networks. AB - The parietal cortex has been proposed as part of the neural network for guiding spatial attention. However, it is unclear to what degree the parietal cortex contributes to the attentional modulations of activities of the visual cortex and the engagement of the frontal cortex in the attention network. We recorded behavioural performance and haemodynamic responses using functional MRI from a patient with focal left parietal damage in covert visual orienting tasks requiring detection of targets at the attended or unattended locations. While the patient's reaction times to left visual field stimuli were speeded by valid relative to invalid cues, attention to LVF stimuli was associated with enhanced activities in the right extrastriate cortex, right parietal and cingulate cortices, and bilateral frontal cortices. However, the patient's behavioural and neural responses to right visual field stimuli were not influenced by cue validity. The results are discussed in terms of the role of human parietal cortex in the neural network underlying voluntary attentional control. PMID- 14761903 TI - What's in a name: voxel-based morphometric analyses of MRI and naming difficulty in Alzheimer's disease, frontotemporal dementia and corticobasal degeneration. AB - Confrontation naming is impaired in neurodegenerative conditions like Alzheimer's disease (AD), frontotemporal dementia (FTD) and corticobasal degeneration (CBD). Some behavioural observations suggest a common source of impaired naming across these patient groups, while others find partially unique patterns of naming difficulty. We hypothesized that a large-scale neural network underlies naming, and that patterns of impaired naming in AD, FTD and CBD reflect cortical atrophy that interrupts this network in a manner that is partially shared and partially unique across these patient groups. We tested this hypothesis by correlating naming impairments with voxel-based morphometric (VBM) analyses of cortical atrophy in structural MRIs of 50 patients. We found significant naming deficits in all patient groups. Naming also correlated with lexical retrieval in all patient groups, including subgroups of patients with FTD. VBM analyses showed significant cortical atrophy, which was shared across AD, FTD and CBD patients in the left lateral temporal cortex; this area correlated with naming accuracy in all groups. Left lateral temporal atrophy thus appears to interfere with a lexical retrieval component of naming in AD, FTD and CBD. Impaired naming also correlated with semantic memory and visual perceptual-spatial functioning in specific groups of patients and, correspondingly, naming correlated with cortical atrophy in partially distinct neuroanatomical distributions in AD, FTD, CBD and subgroups of patients with FTD. These partially unique correlation profiles appear to reflect selective interruption of other components of the naming process, including semantic and visual perceptual-spatial functioning. These findings are consistent with the hypothesis that a large-scale neural network supports naming, and that this network is interrupted in several distinct ways in patients with neurodegenerative diseases. PMID- 14761904 TI - Calpain inhibition protects against Taxol-induced sensory neuropathy. AB - Taxol is a highly effective anticancer agent that causes peripheral neuropathy as its major toxic side effect. The neuropathy is characterized by degeneration of sensory axons that may be severe enough to be dose limiting. Axonal degeneration involves the activation of the calcium-activated proteases calpains, and here we tested whether systemic inhibition of calpains with the peptide alpha-ketoamide calpain inhibitor AK295 can reduce the clinical and pathological effects of Taxol in a rodent model of Taxol neuropathy. In mice with Taxol neuropathy, AK295 reduced the degree of axonal degeneration in sensory nerve roots, and improved clinical measures of neuropathy, including behavioural and electrophysiological function. These findings were consistent for both 3- and 6-week models of neuropathy. In vitro, Taxol caused activation of both calpains and caspases in PC12 cells. AK295 inhibited the activation of calpains but did not interfere with the antimitotic effects of Taxol on microtubules, nor did it inhibit caspase mediated cell death. These data implicate calpains in the pathogenesis of Taxol neuropathy, and demonstrate that AK295 can prevent axonal degeneration and clinical neuropathy in mice. In addition, AK295 did not interfere with the primary antineoplastic effects of Taxol on microtubules and cell death, suggesting that systemic calpain inhibition may be a good strategy for preventing neuropathy in patients being treated with Taxol. PMID- 14761905 TI - Turning a blind eye: the success of blinding reported in a random sample of randomised, placebo controlled trials. AB - OBJECTIVE: To examine the reporting and success of double blinding in a sample of randomised, placebo controlled trials from leading general medicine and psychiatry journals. METHODS: Identification of placebo controlled, randomised controlled trials from prespecified general medical and psychiatric journals indexed on Medline between 1 January 1998 and 1 October 2001, from which a random sample of 200 randomised clinical trials was chosen, of which 191 trials were evaluated. RESULTS: Only seven of the 97 (7%) general medicine trials provided evidence on the success of blinding, with five reporting that the success of blinding was imperfect. In trials from psychiatric journals, the success of blinding was reported in eight of the 94 trials, with four reporting that the blinding was imperfect. Overall, only four of the 191 (2%) trials assessed blinding in the participants and either the outcome assessors or the investigators. CONCLUSIONS: The current lack of reporting on the success of blinding provides little evidence that success of blinding is maintained in placebo controlled trials. Trialists and editors should make a concerted effort to incorporate, report, and publish such information and its potential effect on study results. PMID- 14761906 TI - Cognitive ability in childhood and cognitive decline in mid-life: longitudinal birth cohort study. AB - OBJECTIVE: To examine the association between cognitive ability in childhood and mid-life cognitive decline in the normal population. DESIGN: Longitudinal, population based, birth cohort study. PARTICIPANTS: 2058 men and women born in 1946. MAIN STUDY MEASURES: Ability in childhood measured by AH4 and test of verbal comprehension at age 15 years. Ability in adulthood measured by the national adult reading test (NART) at age 53 years. Outcome measures were decline in memory (word list learning) and speed and concentration (timed visual search) from age 43 to 53 years. RESULTS: Ability in childhood was significantly and negatively associated with decline in memory (beta = 0.09, P = 0.005, for men; 0.10, P < 0.001, for women) and search speed (beta = 0.13, P < 0.001, for men; 0.08, P = 0.01, for women), independent of educational attainment, occupational social class, and a range of health indicators. The adult reading test was also significantly and negatively associated with decline in these outcomes (for memory beta = 0.21, P < 0.001, for men; 0.17, P < 0.001, for women; and for search speed beta = -0.05 for men; 0.10, P = 0.008 for women) independent of educational attainment, social class, and childhood ability. CONCLUSIONS: Ability in childhood can protect against cognitive decline in mid-life and beyond. Results for the adult reading test indicate that the protective effect of ability may also be acquired in adulthood. PMID- 14761908 TI - Society of Surgical Oncology presidential address: volume, outcome, and surgical specialization. PMID- 14761909 TI - Are minimally invasive techniques for ablation of breast cancer ready for "Prime Time"? PMID- 14761910 TI - From bad to worse: prognostic factors in pancreatic cancer. PMID- 14761911 TI - Is there a role for isolated limb perfusion with tumor necrosis factor in patients with melanoma? PMID- 14761912 TI - Peritoneal carcinomatosis or liver metastases from colorectal cancer: similar standards for a curative surgery? PMID- 14761914 TI - Early-stage gastric cancer: a highly treatable disease. PMID- 14761913 TI - Are we overtreating rectal cancer: time for another trial? PMID- 14761915 TI - Stereotactic body radiation therapy: an ablative treatment option for primary and secondary liver tumors. AB - Only a subset of patients with primary and secondary liver tumors are eligible for surgical resection because of either the presence of extrahepatic disease, increased number of hepatic lesions, the anatomical distribution of tumors within the liver, and/or general medical inoperability. Nonsurgical, ablative tumor treatment may benefit selected patients by preserving normal liver function. This review presents the concept and technology of stereotactic body radiation therapy and summarizes available clinical data describing applications in the treatment of malignant liver tumors. We present predominantly peer-reviewed data but also summarize recent clinical developments along with discussions of current ongoing and planned multicenter studies. PMID- 14761916 TI - Focused microwave phased array thermotherapy for ablation of early-stage breast cancer: results of thermal dose escalation. AB - BACKGROUND: Tumor ablation as a means of treating breast cancer is being investigated. Microwave energy is promising because it can preferentially heat high-water-content breast carcinomas, compared to adipose and glandular tissues. METHODS: This is a prospective, multicenter, nonrandomized dose-escalation study of microwave treatment. Thermal dose was measured as (1) thermal equivalent minutes (cumulative equivalent minutes; CEM) of treatment relative to a temperature of 43 degrees C and (2) peak tumor temperature. Microwaves were guided by an antenna-temperature sensor placed percutaneously into the tumor. Outcomes measured were pathologic response (tumor necrosis) side effects. RESULTS: Twenty-five patients (mean age, 57 years) were enrolled. The mean tumor diameter was 1.8 cm. Tumoricidal temperatures (>43 degrees C) were reached in 23 patients (92%). Tumor size was unchanged after thermotherapy (P = not significant). Pathologic necrosis was achieved in 17 (68%) patients. Complete necrosis of the invasive component was achieved in two patients. One hundred forty CEM is predictive of a 50% tumor response, and 210 CEM is predictive of a 100% tumor response (P =.003). Univariate linear regression predicts that peak tumor temperatures of 47.4 degrees C and 49.7 degrees C cause a 50% tumor response and a 100% tumor response, respectively. CONCLUSIONS: Thermotherapy causes tumor necrosis and can be performed safely with minimal morbidity. The degree of tumor necrosis is a function of the thermal dose. Future studies will evaluate the impact of high doses of thermotherapy on margin status and complete tumor ablation. PMID- 14761917 TI - Intratumoral IL-12 and TNF-alpha-loaded microspheres lead to regression of breast cancer and systemic antitumor immunity. AB - BACKGROUND: Local, sustained delivery of cytokines at a tumor can enhance induction of antitumor immunity and may be a feasible neoadjuvant immunotherapy for breast cancer. We evaluated the ability of intratumoral poly-lactic-acid encapsulated microspheres (PLAM) containing interleukin 12 (IL-12), tumor necrosis factor alpha (TNF-alpha), and granulocyte-macrophage colony stimulating factor (GM-CSF) in a murine model of breast cancer to generate a specific antitumor response. METHODS: BALB/c mice with established MT-901 tumors underwent resection or treatment with a single intratumoral injection of PLAM containing IL 12, TNF-alpha, or GM-CSF, alone or in combination. Two weeks later, lymph nodes and spleens were harvested, activated with anti-CD3 monoclonal antibodies (mAb) and rhIL-2, and assessed for antitumor reactivity by an interferon gamma (IFNgamma) release assay. Tumor-infiltrating lymphocyte (TIL) analysis was performed on days 2 and 5 after treatment by mechanically processing the tumors to create a single cell suspension, followed by three-color fluorescence activated cell sorter (FACS) analysis. RESULTS: Intratumoral injection of cytokine-loaded PLAM significantly suppressed tumor growth, with the combination of IL-12 and TNF-alpha leading to increased infiltration by polymorphonuclear cells and CD8+ T-cells in comparison with controls. The induction of tumor specific reactive T-cells in the nodes and spleens, as measured by IFN-gamma production, was highest with IL-12 and TNF-alpha. This treatment resulted in resistance to tumor rechallenge. CONCLUSIONS: A single intratumoral injection of IL-12 and TNF-alpha-loaded PLAM into a breast tumor leads to infiltration by polymorphonuclear cells and CD8+ T-cells with subsequent tumor regression. In addition, this local therapy induces specific antitumor T-cells in the lymph nodes and spleens, resulting in memory immune response. PMID- 14761918 TI - Survival and recurrence after breast cancer in BRCA1/2 mutation carriers. AB - BACKGROUND: Genetic mutation is responsible for approximately 10% of breast cancers. The purpose of this study was to compare breast cancer survival and recurrence rates between BRCA1/2 mutation carriers and noncarriers. METHODS: Using the Columbia Presbyterian breast cancer database, we collected the tissue blocks of all patients younger than 65 years of age and of Jewish descent. The patients were contacted and the data updated. DNA was extracted from the tissue blocks and tested for the common mutations. The results of the genetic mutation and updated database were anonymized and merged. The survival and recurrence rates were compared between mutation carriers and noncarriers. RESULTS: A total of 739 breast cancer cases in 715 patients were identified. We were able to test 487 patients. We identified 30 BRCA1 and 21 BRCA2 mutation carriers, for an incidence of 10.36%. The median follow-up for the patients tested was 50 months. BRCA1 patients more frequently had estrogen- and progesterone-negative tumors and had a higher incidence of positive nodes. BRCA1 patients received chemotherapy more frequently. The incidence of in situ disease was similar for mutation and non-mutation carriers. BRCA1/2 mutation carriers had a higher incidence of bilateral disease. There was no difference in 5- or 10-year overall and breast cancer-specific survival between mutation and non-mutation carriers. CONCLUSIONS: Breast cancer patients with BRCA1/2 mutations have a similar outcome as non mutation carriers. PMID- 14761919 TI - Increased expression of valosin-containing protein (p97) is associated with lymph node metastasis and prognosis of pancreatic ductal adenocarcinoma. AB - BACKGROUND: Valosin-containing protein (VCP, also known as p97) exhibits antiapoptotic function and metastasis by activation of nuclear factor kappa-B signaling pathway. Our previous study showed that VCP expression level correlated with prognosis of hepatocellular and gastric carcinoma. In the present study, association of VCP expression with lymph node metastasis and prognosis of pancreatic ductal adenocarcinoma (PDAC) was examined. METHODS: VCP expression in 83 patients (46 males and 37 females) of ages ranging from 43 to 80 (median, 66) years who had undergone curative surgery for primary PDAC was analyzed by immunohistochemistry, in which staining intensity in tumor cells was categorized as weaker or equal to (low expression) or stronger (high expression) than that in noncancerous ductal tissue. RESULTS: Thirty-two tumors (38.6%) and 51 tumors (61.4%) were classified as low-VCP-expressing and high-VCP-expressing tumors, respectively. VCP expression correlated significantly with lymph node metastasis (P <.01) but not with various clinicopathologic factors, including age, gender, and histologic differentiation. Multivariate analysis revealed VCP expression as an independent prognosticator for both disease-free and overall survival, along with histologic differentiation, T stage of pathologic tumor-node-metastasis (pTNM) classification, and lymph node metastasis. Furthermore, VCP expression was a prognosticator for disease-free and overall survival in each relatively early stage (I or II) and advanced stage (III) group of pTNM classification. CONCLUSIONS: Our results indicate the potential usefulness of VCP expression as a marker of metastasis and overall prognosis of PDAC. PMID- 14761920 TI - Hyperthermic isolated limb perfusion with low-dose tumor necrosis factor-alpha and melphalan for bulky in-transit melanoma metastases. AB - BACKGROUND: Melphalan (L-PAM) hyperthermic isolated limb perfusion (HILP) is currently considered the standard treatment for patients with in-transit metastases from cutaneous melanoma. We here report on the results of L-PAM and low-dose tumor necrosis factor (TNF)alpha HILP in patients with bulky disease. METHODS: Twenty patients underwent TNFalpha (1 mg) and L-PAM (10 mg/L) HILP. Perfusion was performed for 90 minutes, and systemic leakage was strictly monitored. Locoregional toxicity was evaluated according to Wieberdink's criteria, whereas tumor response was evaluated with physical examination and ultrasound scan with or without fine-needle aspiration of any suspected recurrence. RESULTS: In all cases, systemic leakage was <5%. No postoperative deaths occurred, and locoregional toxicity was mild (grade 1 or 2) in 95% of patients. A complete tumor response was obtained in 14 patients (70%), and partial responses were obtained in 5 patients (25%). After a median follow-up of 18 months, six patients are alive and disease free, seven are alive with local or distant recurrence or both, and seven have died of disease. CONCLUSIONS: Low-dose TNFalpha HILP can achieve tumor responses comparable with those reported with higher doses of cytokine. Moreover, this drug regimen is associated with acceptable local toxicity, carries a smaller risk of systemic toxicity, and incurs lower costs. PMID- 14761921 TI - Cytoreductive surgery and intraperitoneal hyperthermic chemotherapy with mitomycin C for peritoneal carcinomatosis from nonappendiceal colorectal carcinoma. AB - BACKGROUND: Cytoreductive surgery (CS) and intraperitoneal hyperthermic chemotherapy (IPHC) are efficacious in patients with disseminated mucinous tumors of the appendix. We reviewed our experience using this approach for nonappendiceal colorectal cancer (NACC). METHODS: We performed a retrospective chart review of a prospective database for patients undergoing CS and IPHC with mitomycin C for peritoneal carcinomatosis from colorectal primary lesions between December 1991 and April 2002. RESULTS: There were 77 patients, with a median age of 54 years. Peritoneal carcinomatosis was synchronous and metachronous in 27% and 73% patients, respectively. Seventy-five percent of patients (n = 58) had received chemotherapy prior to IPHC. Complete resection of all gross disease was accomplished in 37 patients (48%). The mean carcinoembryonic antigen level decreased from a preoperative value of 31.2 to a postoperative value of 6.9 (P <.0001). Overall survival (OS) at 1, 3, and 5 years was 56%, 25%, and 17%, respectively. With a median follow-up of 15 months, the median OS was 16 months. Perioperative morbidity and mortality were 30% and 12%, respectively. Hematologic toxicity occurred in 15 patients (19%). Cox regression analysis identified poor performance status (P =.018), bowel obstruction (P =.001), malignant ascites (P =.001), and incomplete resection of gross disease (P =.011) as independent predictors of decreased survival. Patients with complete resection of all gross disease had a 5-year OS of 34%, with a median OS of 28 months. CONCLUSIONS: CS and IPHC with mitomycin C can improve outcomes for select patients with peritoneal spread from NACC. One third of patients who undergo complete resection of gross disease have long-term survival. PMID- 14761922 TI - Residual mesorectal lymph node involvement following neoadjuvant combined modality therapy: rationale for radical resection? AB - BACKGROUND: In order to evaluate the impact of preoperative radiation and chemotherapy (combined modality therapy, or CMT) on primary rectal cancer and mesorectal lymph nodes (MLNs), middle and lower third rectal cancers were resected with total mesorectal excision (TME) and assessed for frequency of MLN retrieval and residual MLN involvement. METHODS: Between 1990 and 2001, 187 consecutive patients underwent abdominoperineal resection (APR) or low anterior resection (LAR) for locally advanced (endorectal ultrasound [ERUS] stage, T3-4) mid and distal rectal cancer following preoperative CMT. Sphincter preservation was possible in 150 patients (80%). The mean number of retrieved MLNs was 10.6. Pre-CMT ERUS stage was compared with final pathologic stage. RESULTS: Comparison of pre-CMT ERUS stage with pathologic stage revealed a decrease in T stage in 93 patients (49%), as well as a decrease in the percentage of individuals with positive MLNs, from 54% to 27% (P <.0001). The overall incidence of positive MLN involvement was 27%, and incidence paralleled pathologic T stage (pT): pT0 = 7%, pT1 = 8%, pT2 = 22%, pT3 = 37%, and pT4 = 67%. CONCLUSIONS: Following preoperative CMT, the incidence of residual MLN involvement remains significant and parallels increasing pT stage. Therefore, the standard of care for locally advanced distal rectal cancer should continue to include formal rectal resection (TME). PMID- 14761923 TI - Deleted in oral cancer-1 expression upregulates proapoptosis elements in microsatellite-unstable human colorectal cancer. AB - BACKGROUND: We previously reported differential expression of the growth suppressor, deleted in oral cancer-1 (DOC-1), in microsatellite-unstable (MSI+) versus microsatellite-stable colorectal cancer (CRC) cell lines. MSI+ CRC cell lines demonstrated decreased DOC-1 expression and decreased apoptosis. Transfection of wild-type DOC-1 into an MSI+ cell line (SW48) resulted in increased apoptosis. We undertook our current experiment to identify specific elements modulated by DOC-1 expression that result in increased apoptosis. METHODS: SW48 is an MSI+ CRC cell line that does not constitutively express DOC 1. SW48 was suspended in culture medium and incubated to 60% confluence. Half the plates were transfected with cytomegalovirus (CMV)-DOC-1. At 30 hours, RNA and protein were isolated with Trizol. Complementary DNA microarray was performed to compare SW48(CMV-DOC-1) with SW48, which lacks DOC-1. Signal intensity was analyzed by GenePix Pro 3.0 software. Expression ratios / 1.5 were considered significant. Poor-quality spots were flagged and excluded from analysis. Real-time polymerase chain reaction was performed to determine DOC-1 levels in both cell lines. RESULTS: Successful transfection of DOC-1 was confirmed by real-time polymerase chain reaction and by Western blot. Microarray revealed significant differential expression of DOC-1, as expected. Increased DOC 1 expression in SW48(CMV-DOC-1) was associated with significantly increased expression of proapoptosis components of the caspase cascade (CASP7, CASP9) and bcl2/bax pathway (BNIP3, BNIP3L, BID). CONCLUSIONS: DOC-1 expression promotes apoptosis by upregulation of specific elements of the caspase cascade and bcl2/bax pathways. DOC-1 therefore deserves further study as a candidate for the therapeutic modulation of apoptosis in MSI+ CRC. PMID- 14761924 TI - Pilot study using a humanized CC49 monoclonal antibody (HuCC49DeltaCH2) to localize recurrent colorectal carcinoma. AB - BACKGROUND: CC49 is a monoclonal antibody directed against a pancarcinoma antigen (TAG-72) expressed by colorectal cancers. The use of murine CC49 in radioimmunoguided surgery (RIGS) was problematic because of the human anti-mouse antibodies (HAMA) generated. This study was designed to assess the clearance, safety, and effectiveness of localization of a complimentarity determining region (CDR)-grafted humanized domain-deleted antitumor CC49 antibody (HuCC49DeltaCH2). METHODS: After thyroid blockade, 1 mg of HuCC49DeltaCH2 radiolabeled with 2 mCi of iodine-125 was administered. All patients subsequently underwent traditional exploration followed by a survey with the gamma-detecting probe. In five patients, exploration was performed 10 to 24 days after injection, when precordial counts were sufficiently low (<30 counts per 2 seconds [cp2s]). Traditionally suggestive and probe-positive tissue was biopsied or excised and examined for the presence of carcinoma, when considered appropriate by the operating surgeon. Serum was assessed for HAMA. RESULTS: Seventeen sites were identified as suggestive of carcinoma on traditional exploration and 21 by RIGS. Of these, pathologic correlation was obtained in 15. The sensitivity of RIGS was 92%, and the positive predictive value was 100%. None of the patients expressed significant HAMA. CONCLUSIONS: This initial study indicates that the HuCC49DeltaCH2 monoclonal antibody, when used with RIGS, is safe and sensitive in detecting recurrent intra-abdominal colon cancer. PMID- 14761925 TI - Comparative evaluation of gastric carcinoma staging: Japanese classification versus new american joint committee on cancer/international union against cancer classification. AB - BACKGROUND: The TNM and Japanese classifications of regional lymph node spread (N categories) for gastric cancer differ, whereas the classifications of local extent (T categories) are identical. This study was designed to compare these staging systems and devise a more rational system for gastric carcinoma. METHODS: A series of 1244 patients with gastric cancer were enrolled in the study. Survival rates were evaluated to clarify which aspects of each staging system (feasibility, reproducibility, and accuracy of prognostic stratification) were superior. RESULTS: The TNM and Japanese classification systems differ in their categorizations of lymph node spread. A significant difference in survival rate was observed in lymph node metastasis classified as N1 and N2 by the Japanese classification and then subclassified by the TNM classification, although there was no significant difference in the survival in cases of lymph node metastasis classified by TNM into pN1 and pN2 and then subclassified by the Japanese classification. Among patients with M1 metastasis (number 16 a2 and b1 in Japanese classification) in the TNM classification, there was a significant difference in survival. A new classification that included the para-aortic lymph nodes (number 16 a2 and b1) as regional lymph nodes within the TNM classification provided homogeneity and an improvement in staging. CONCLUSIONS: TNM classification was more rational and homogenous than Japanese classification. New classification could lead to worldwide uniformity in the description of patients and make possible uniform interinstitutional comparisons of surgical results. PMID- 14761926 TI - Percutaneous imaging-guided radiofrequency ablation in patients with colorectal pulmonary metastases: 1-year follow-up. AB - BACKGROUND: We assessed the safety and evidence of efficacy of radiofrequency ablation (RFA) for colorectal lung metastases with follow-up to 1 year. METHODS: Twenty-three patients had percutaneous RFA for 52 colorectal pulmonary metastases under fluoro-computed tomography (CT). Patients received intravenous conscious sedation and local analgesia with routine hospitalization and monitoring for 24 hours after RFA. Patients had CT scanning at 1 month and then every 3 months, with serum carcinoembryonic antigen assessment monthly and every 3 months. RESULTS: All ablations were technically successful. Tumor diameter ranged from.3 to 4.2 cm. Pneumothorax occurred in 43% (10 of 23) of patients. Six patients required intercostal chest drain placement. Six patients had a second RFA, four for new lesions and two for re-treatment of a previously treated lesion. The median admission was 2.0 days (range, 1-9 days). The median follow-up was 428 days (range, 173-829 days); data are reported to 1 year in this article. Five patients died at 5, 6, 8, 8, and 12 months after RFA from extrapulmonary (n = 1) or widespread (n = 4) disease. One patient developed a malignant pleural effusion at 6 months after RFA. Cavitation was seen in nine treated lesions (17%); all resolved with scar tissue contraction by 12 months. Eighteen patients with CT scan follow-up at 1 year have 40 lesions classified as disappeared (n = 17), decreased (n = 5), stable/same size (n = 4), or increased (n = 14). CONCLUSIONS: Percutaneous imaging-guided RFA of multiple colorectal pulmonary metastases is a minimally invasive treatment option with modest morbidity. A significant proportion of patients show good evidence of successful local control at 1 year. PMID- 14761927 TI - Improved staging of cervical metastases in clinically node-negative patients with head and neck squamous cell carcinoma. AB - BACKGROUND: The management of the N0 neck in oral and oropharyngeal cancer is often determined by the risk of metastases related to features of the primary tumor. Where the risk of metastases is >20%, elective neck dissection (END) has been advocated. This study reviewed clinical staging, surgical staging, pathologic staging, and histopathologic parameters to determine the prediction of nodal metastases and micrometastases in patients with head and neck squamous cell carcinoma. METHODS: A prospective series of 61 clinically neck node-negative patients undergoing surgical resection of a T1/2 intraoral or oropharyngeal invasive squamous cell carcinoma and surgical staging of the neck, with sentinel node biopsy (SNB) alone or SNB-assisted END, between June 1998 and March 2002 were included in this study. RESULTS: Pathologic upstaging of the clinically N0 neck occurred in 27 (44%) of 61 patients. Routine pathology with hematoxylin and eosin upstaged disease in 22 of 27 patients (sensitivity of 81%). Five patients with micrometastasis were staged pN1mi after stepped serial sectioning and immunohistochemistry. Tumor thickness, a noncohesive invasive front, and perineural and bone invasion were all histological predictors for cervical metastases. Five patients with micrometastases were staged pN1mi. CONCLUSIONS: Both clinical staging and routine pathologic staging underestimate the presence of nodal metastases. Staging with either SNB alone or SNB-assisted END shows promise in the management of the N0 neck by identifying patients with micrometastases (pN1mi). PMID- 14761928 TI - Perioperative morbidity and mortality in elderly gynecological oncological patients (>/= 70 Years) by the American Society of Anesthesiologists physical status classes. AB - BACKGROUND: We evaluated the morbidity and mortality associated with American Society of Anesthesiologists (ASA) classes III and IV versus ASA classes I and II in elderly women (>/= 70 years) undergoing gynecological oncological surgery. METHODS: From 1986 to 2000, we retrospectively collected patients >/= 70 years of age undergoing oncological gynecological surgery. The study population consisted of 121 ASA class III and IV patients. The control group consisted of the same number of patients with ASA classes I and II, and these were matched to study patients (1:1) by clinical and surgical data. The morbidity and mortality of patients with ASA status III and IV were analyzed before and after 1992. RESULTS: In ASA class III and IV patients, compared with ASA class I and II, a higher rate of severe morbidity (P =.000) occurred, whereas the median postoperative stay was similar (8 days). No differences between patients with ASA class III and IV and ASA class I and II for median operative time, transfusion rate, or median blood loss were found. Mortality was 3% in ASA classes III and IV. CONCLUSIONS: Our study suggests that surgery in elderly gynecological oncological patients aged >/= 70 years with ASA class III or IV results in an acceptable perioperative morbidity and mortality rate. PMID- 14761929 TI - Operative surgical education: results of a society of surgical oncology fellowship survey and proposal for an operative database. AB - BACKGROUND: Preparation of surgical trainees for oncological operative procedures is challenging. The purpose of this survey was to focus on identifying methods and resources used by trainees to prepare for procedures and to ascess the need for additional educational tools. METHODS: A 34-item survey was mailed electronically to 97 surgical oncology fellows at 14 Society of Surgical Oncology approved training programs. General surgery residents at an affiliate training program (n = 65) and residents attending an American Board of Surgery In-Training Examination review course (n = 129) were polled via hard-copy mailings. The survey was distributed with the Dillman method. Self-education practices, factors influencing operative education, and strengths/weaknesses of available resources were identified. RESULTS: Response rates were 56% and 78% for fellows and residents, respectively. Trainees prepare for more than 50% of cases they perform (82%; 169 of 205), devoting up to 1 hour (87%; 178 of 205) in review the evening before a procedure (64%; 131 of 205). Time availability and attending of record were dominant factors influencing resident preparation, whereas case complexity was the most important variable motivating fellows. Surgical atlases, texts, anatomical references, and case discussion with attending staff were the most useful and available resources rated by trainees. Skills stations were recognized as the least valuable. Critical assessment of six educational resources identified no one particular area for improvement. CONCLUSIONS: There is a need for contemporary operative educational tools, incorporating time-sensitive and procedure-specific needs of surgical trainees preparing for oncological operative procedures. PMID- 14761930 TI - The story so far: Molecular regulation of the heme oxygenase-1 gene in renal injury. AB - Heme oxygenases (HOs) catalyze the rate-limiting step in heme degradation, resulting in the formation of iron, carbon monoxide, and biliverdin, the latter of which is subsequently converted to bilirubin by biliverdin reductase. Recent attention has focused on the biological effects of product(s) of this enzymatic reaction, which have important antioxidant, anti-inflammatory, and cytoprotective functions. Two major isoforms of the HO enzyme have been described: an inducible isoform, HO-1, and a constitutively expressed isoform, HO-2. A third isoform, HO 3, closely related to HO-2, has also been described. Several stimuli implicated in the pathogenesis of renal injury, such as heme, nitric oxide, growth factors, angiotensin II, cytokines, and nephrotoxins, induce HO-1. Induction of HO-1 occurs as an adaptive and beneficial response to these stimuli, as demonstrated by studies in renal and non-renal disease states. This review will focus on the molecular regulation of the HO-1 gene in renal injury and will highlight the interspecies differences, predominantly between the rodent and human HO-1 genes. PMID- 14761931 TI - Microalbuminuria as a marker of cardiovascular and renal risk in type 2 diabetes mellitus: a temporal perspective. AB - Microalbuminuria is a marker for diabetic nephropathy. It also signifies cardiovascular disease, as well as nephropathy, in type 2 diabetes (DM2). Microalbuminuria may precede DM2, occurring with the insulin resistance syndrome and its components, including obesity and hypertension. Other indicators of cardiovascular risk, such as markers of inflammation, are associated with microalbuminuria in populations of patients with and without diabetes. With the rising prevalence of DM2 in minority youth, especially in Native Americans, a marker for future disease risk would allow earlier prevention strategies to be tested. Before microalbuminuria can be used in a prevention strategy, more needs to be known about the mechanism(s) of the association between elevated excretion, its relationship to glucose intolerance, and its relative contribution to cardiovascular and renal disease. These questions are especially applicable as we begin to observe the long-term complications of diabetes in youth. PMID- 14761932 TI - Reduced GRK2 level in T cells potentiates chemotaxis and signaling in response to CCL4. AB - Chemokine receptors belong to the family of G-protein-coupled receptors (GPCR). Phosphorylation of GPCR by GPCR kinases (GRKs) is considered to play an important role in desensitization of these receptors. We have recently shown in patients with rheumatoid arthritis that the level of GRK2 in lymphocytes is reduced by approximately 50%. However, the physiological relevance of reduced GRK2 levels in lymphocytes is not known. Here, we investigated whether reduced GRK2 expression changes the chemotactic response of T cells to the chemokines CCL3, CCL4, and CCL5. Activated T cells from GRK2+/- mice, which have a 50% reduction in GRK2 protein levels, showed a significant 40% increase in chemotaxis toward the CCR5 ligand CCL4. In addition, chemotaxis toward the CCR1 and CCR5 ligands CCL3 and CCL5 was also increased. Binding of CCL4 to activated T cells from GRK2+/- and wild-type (WT) mice was similar, but agonist-induced CCR5 phosphorylation was attenuated in GRK2+/- cells. Moreover, the calcium response and phosphorylation of protein kinase B and extracellular-regulated kinase in response to CCL4 were significantly increased in GRK2+/- T cells, showing that signaling is increased when the level of GRK2 is reduced. GRK2+/- and WT cells do become refractory to restimulation with CCL4. In conclusion, a 50% decrease in T cell GRK2 expression results in increased responsiveness to CCL3, CCL4, and CCL5, suggesting that the 50% reduction in lymphocyte GRK2 level as observed during inflammation can have functional consequences for the response of these cells to chemokines. PMID- 14761933 TI - Identification of caspase-10 in human neutrophils and its role in spontaneous apoptosis. AB - In the present study, we investigated the molecular mechanisms of spontaneous and tumor necrosis factor alpha (TNF-alpha)-mediated apoptosis of human polymorphonuclear neutrophils (PMN). Whereas TNF-alpha-mediated apoptosis was almost absent in the presence of the caspase-8 inhibitor Z-Ac-Ala-Glu-Val-Asp-7 fluoromethyl ketone (Z-AEVD-FMK), the inhibitor had no effect on spontaneous apoptosis, suggesting that spontaneous apoptosis was independent of caspase-8. Subsequently, we identified different isoforms of caspase-10 in human PMN and found high expression of caspase-10/b and/or -10/d and low expression of caspase 10/a and -10/c at the mRNA level. At the protein level, freshly isolated PMN showed high expression of caspase-10/b and -10/d as well as moderate expression of caspase-10/a and -10/c. Upon spontaneous apoptosis, caspase-10/b was down regulated, which was accompanied by the appearance of a specific 47-kDa caspase 10/b cleavage product and an increased caspase-10 activity. In contrast, no down regulation of caspase-10/a, -10/c, or -10/d was observed, suggesting that spontaneous apoptosis was associated with a differential activation of caspase 10/b. This was confirmed by the finding that spontaneous apoptosis was inhibited in the presence of Z-Ile-Glu-Thr-Asp (Z-IETD)-FMK, which blocks caspase-10. However, no down-regulation of caspase-10 isoforms was observed in the presence of TNF-alpha, suggesting that caspase-10 was not involved in TNF-alpha-induced apoptosis. Taken together, our study demonstrates that spontaneous and TNF-alpha mediated apoptosis of PMN have different molecular requirements. Whereas TNF alpha-mediated apoptosis depends on the activation of caspase-8, spontaneous apoptosis requires the activation of caspase-10/b. This finding may reveal that PMN apoptosis in different (patho-) physiological settings results from distinct molecular mechanisms. PMID- 14761934 TI - Mannose-binding lectin deficiency alters the development of fungal asthma: effects on airway response, inflammation, and cytokine profile. AB - Aspergillus fumigatus is a major fungal pathogen that may be fatal to immunocompromised individuals and causes airway hyperreactivity and remodeling in sensitized individuals. Herein, we examined the role of mannose-binding lectin (MBL), a complement-activating plasma protein, during pulmonary innate and allergic immune responses directed against A. fumigatus spores or conidia. Neither group of nonsensitized MBL-A-sufficient (MBL-A+/+) nor -deficient (MBL-A /-) mice challenged with an intravenous or intratracheal (i.t.) bolus of A. fumigatus spores experienced fungus-induced mortality, but marked airway remodeling was observed in MBL-A-/- mice challenged i.t. with conidia. In a model of chronic fungal asthma, MBL-A+/+ and MBL-A-/- A. fumigatus-sensitized mice were examined at days 4 and 28 after an i.t. challenge with A. fumigatus conidia. Airway hyperresponsiveness in sensitized MBL-A-/- mice was significantly decreased at both times after conidia challenge compared with the sensitized MBL A+/+ group. In the sensitized MBL-A-/- mice, whole lung T helper cell type 2 cytokine levels were significantly decreased at day 4 after conidia, and whole lung interferon-gamma levels were significantly increased at day 28 after conidia when compared with controls. However, histological evidence showed similar airway remodeling at day 28 after conidia (i.e., subepithelial fibrosis and goblet cell metaplasia) in the two groups of mice. Thus, these findings show that MBL-A is not required for mouse survival following exposure to A. fumigatus conidia, and this murine collectin isoform contributes to the development and maintenance of airway hyperresponsiveness but not chronic airway remodeling during chronic fungal asthma. PMID- 14761935 TI - Differential gene expression profile of human tonsil high endothelial cells: implications for lymphocyte trafficking. AB - Lymphocyte recirculation is dependent on the interactions of adhesion and signaling molecules expressed on lymphocytes and their partners on high endothelial cells (HEC). Many of the events in this process have yet to be molecularly characterized. To identify novel HEC-specific proteins with potential function in the recruitment cascade, we sequenced a normalized human tonsil HEC cDNA library (generated from an inflamed tonsil) from which lymphocyte and human umbilical vein endothelial cell cDNAs had been subtracted. One-thousand forty nine sequences were analyzed. All but three mapped to known cDNAs or genomic DNAs. The two most abundant transcripts encoded alpha2-macroglobulin and hevin. The next-abundant transcripts encoded several other protease inhibitors, making this protein class the most prominent in HEC. Several endothelial-specific transcripts were also identified, including those encoding E-selectin, vascular cell adhesion molecule-1, vascular endothelial-junctional adhesion molecule, and platelet-endothelial cell adhesion molecule-1. The library contains a great diversity of transcripts, and studies of the encoded proteins will provide further insight into the complex biology of these specialized endothelial cells. PMID- 14761936 TI - Immature dendritic cells reduce proinflammatory cytokine production by a coculture of macrophages and apoptotic cells in a cell-to-cell contact-dependent manner. AB - We have demonstrated that phagocytosis of late apoptotic cells by mouse macrophages leads to the production of proinflammatory cytokines, notably macrophage-inflammatory protein (MIP-2), and therefore, a yet-unknown mechanism(s) should keep our body free of inflammation. In this study, we examined the effect of the addition of immature dendritic cells (iDCs) to a coculture of macrophages and apoptotic cells on MIP-2 production and phagocytosis by macrophages. The addition of iDCs to the coculture reduced MIP-2 production significantly but unexpectedly enhanced the phagocytosis by macrophages. Further study revealed that the reduction of MIP-2 production was dependent on cell-to cell contact partly involving the beta(2) integrin family Mac-1. In addition, anti-inflammatory cytokines, interleukin-10 and transforming growth factor-beta, were involved in the reduction of MIP-2 production, as antibodies against these cytokines recovered MIP-2 production. Both cytokines were expressed by iDCs more significantly than macrophages at the mRNA levels, although they were hardly detected in the supernatant at the protein levels, suggesting that minute amounts of these anti-inflammatory cytokines were produced mainly by iDCs to block MIP-2 production in a cell-to-cell contact-dependent manner. Thus, this study reveals a new mechanism by which MIP-2 production by macrophages phagocytosing apoptotic cells is prevented. PMID- 14761937 TI - Mechanisms of glucocorticoid-induced down-regulation of neutrophil L-selectin in cattle: evidence for effects at the gene-expression level and primarily on blood neutrophils. AB - One anti-inflammatory action of glucocorticoids is down-regulation of surface L selectin on circulating neutrophils. However, it is unclear if this is a result of release of affected bone marrow neutrophils or if the steroid has direct effects on L-selectin expression in existing blood neutrophils. We recently demonstrated that circulating neutrophils from cattle with high blood concentrations of endogenous glucocorticoid had reduced L-selectin mRNA, suggesting that the steroid interrupted L-selectin gene expression. In the current study, dexamethasone (DEX) was administered to cattle in vivo, and blood and bone marrow neutrophils were studied simultaneously within the animal to determine which pool of cells responds to glucocorticoids with inhibited L selectin expression. Purified blood neutrophils were also treated with DEX +/- RU486 in vitro, and glucocorticoid effects on L-selectin expression were determined. Our results indicate that glucocorticoid-induced suppression of L selectin, which accompanies neutrophilia, is likely mediated by direct effects of glucocorticoid receptor activation on intracellular reservoirs of L-selectin mRNA and protein in cattle, predominantly in blood neutrophils. PMID- 14761938 TI - Estrogen replacement, aging, and cell-mediated immunity after injury. AB - We recently demonstrated that aged mice are less likely to survive following traumatic injury and are more immunosuppressed than young mice who sustain comparable injuries. Immunosuppression in severely injured patients and in rodent models of burn injury is associated with a marked elevation in proinflammatory cytokines, including interleukin-6 (IL-6). We reported that after sustaining a moderate-size scald injury, aged mice have higher circulating levels of IL-6 than young, injured mice. As proestrus levels of estrogen have been reported to boost immune responses and attenuate IL-6 production, in the present study, we went on to determine if estrogen replacement in aged female mice restored cellular immunity and proinflammatory cytokine production. After injury, in placebo treated, aged animals, there was a >75% suppression in the delayed-type hypersensitivity response relative to placebo-treated, sham-injured, aged mice (P<0.05). In contrast, estrogen supplementation before injury yielded a partial recovery in this response, such that the mice were suppressed by only 40% relative to sham-injured, aged mice (P<0.05). There was a fourfold increase in the circulating level of IL-6 in burn-injured, aged mice who received placebo hormone replacement relative to sham-injured mice given placebo (P<0.05). This level of cytokine was lowered by nearly 50% in aged, estrogen-treated mice. Most remarkably, estrogen replacement improved survival from 42% (in the absence of estrogen) to 70% in aged, burn-injured mice. Further investigation will be needed to determine if age- and gender-specific therapies are needed for the treatment of all trauma patients. PMID- 14761939 TI - Binding of destabilized betaB2-crystallin mutants to alpha-crystallin: the role of a folding intermediate. AB - Age-related changes in protein-protein interactions in the lens play a critical role in the temporal evolution of its optical properties. In the relatively non regenerating environment of the fiber cells, a critical determinant of these interactions is partial or global unfolding as a consequence of post translational modifications or chemical damage to individual crystallins. One type of attractive force involves the recognition by alpha-crystallins of modified proteins prone to unfolding and aggregation. In this paper, we explore the energetic threshold and the structural determinants for the formation of a stable complex between alpha-crystallin and betaB2-crystallin as a consequence of destabilizing mutations in the latter. The mutations were designed in the framework of a folding model that proposes the equilibrium population of a monomeric intermediate. Binding to alpha-crystallin is detected through changes in the emission properties of a bimane fluorescent probe site-specifically introduced at a solvent exposed site in betaB2-crystallin. alpha-Crystallin binds the various betaB2-crystallin mutants, although with a significantly lower affinity relative to destabilized T4 lysozyme mutants. The extent of binding, while reflective of the overall destabilization, is determined by the dynamic population of a folding intermediate. The existence of the intermediate is inferred from the biphasic bimane emission unfolding curve of a mutant designed to disrupt interactions at the dimer interface. The results of this paper are consistent with a model in which the interaction of alpha-crystallins with substrates is not solely triggered by an energetic threshold but also by the population of excited states even under favorable folding conditions. The ability of alpha-crystallin to detect subtle changes in the population of betaB2 crystallin excited states supports a central role for this chaperone in delaying aggregation and scattering in the lens. PMID- 14761940 TI - The Rsp5 ubiquitin ligase binds to and ubiquitinates members of the yeast CIN85 endophilin complex, Sla1-Rvs167. AB - Sla1 and Rvs167 are yeast proteins required for receptor internalization and organization of the actin cytoskeleton. Here we provide evidence that Sla1 and Rvs167 are orthologues of the mammalian CIN85 and endophilin proteins, respectively, which are required for ligand-stimulated growth factor receptor internalization. Sla1 is similar in domain structure to CIN85 and binds directly to the endophilin-like Rvs167. Akin to CIN85, Sla1 interacts with synaptojanins and a ubiquitin ligase that regulates endocytosis. This ubiquitin ligase, Rsp5, binds directly to both Sla1 and Rvs167. The interaction between Rsp5 and Rvs167 is mediated through Rsp5 WW domains and PXY motifs in the central Gly-Pro-Ala rich domain of Rvs167. Rvs167 PXY motifs are required for Rsp5-dependent monoubiquitination of Rvs167 on Lys481 in the Src homology 3 (SH3) domain. Mutation of Lys481 --> Arg causes cells to grow slowly on medium containing 1 M NaCl, although this phenotype is not due to the defect in ubiquitination caused by the K481R mutation. We propose that Rsp5 interaction with Sla1-Rvs167 promotes Rvs167 ubiquitination and regulates activity of this protein complex. Rvs167 ubiquitination is not required for general function of Rvs167, but may control specific Rvs167 SH3 domain-protein interactions or negatively regulate SH3 domain activity. PMID- 14761941 TI - The Yersinia pseudotuberculosis cytotoxic necrotizing factor (CNFY) selectively activates RhoA. AB - The cytotoxic necrotizing factors (CNF)1 and CNF2 from pathogenic Escherichia coli strains activate RhoA, Rac1, and Cdc42 by deamidation of Gln63 (RhoA) or Gln61 (Rac and Cdc42). Recently, a novel cytotoxic necrotizing factor termed CNFY was identified in Yersinia pseudotuberculosis strains (Lockman, H. A., Gillespie, R. A., Baker, B. D., and Shakhnovich, E. (2002) Infect. Immun. 70, 2708-2714). We amplified the cnfy gene from genomic DNA of Y. pseudotuberculosis, cloned and expressed the recombinant protein, and studied its activity. Recombinant GST-CNFY induced morphological changes in HeLa cells and caused an upward shift of RhoA in SDS-PAGE, as is known for GST-CNF1 and GST-CNF2. Mass spectrometric analysis of GST-CNFY-treated RhoA confirmed deamidation at Glu63. Treatment of RhoA, Rac1, and Cdc42 with GST-CNFY decreased their GTPase activities, indicating that all of these Rho proteins could serve as substrates for GST-CNFY in vitro. In contrast, RhoA, but not Rac or Cdc42, was the substrate of GST-CNFY in culture cells. GST CNFY caused marked stress fiber formation in HeLa cells after 2 h. In contrast to GST-CNF1, formation of filopodia or lamellipodia was not induced with GST-CNFY. Accordingly, effector pull-down experiments with lysates of toxin-treated cells revealed strong activation of RhoA but no activation of Rac1 or Cdc42 after 6 h of GST-CNFY-treatment. Moreover, in rat hippocampal neurons, GST-CNFY results in the retraction of neurites, indicating RhoA activation. In contrast, no activation of Rac or Cdc42 was found. Altogether, our data suggest that CNFY from Y. pseudotuberculosis is a strong, selective activator of RhoA, which can be used as a powerful tool for constitutive RhoA activation without concomitant activation of Rac1 or Cdc42. PMID- 14761942 TI - The effect of disease-associated mutations on the folding pathway of human prion protein. AB - Propagation of transmissible spongiform encephalopathies is believed to involve the conversion of cellular prion protein, PrP(C), into a misfolded oligomeric form, PrP(Sc). An important step toward understanding the mechanism of this conversion is to elucidate the folding pathway(s) of the prion protein. We reported recently (Apetri, A. C., and Surewicz, W. K. (2002) J. Biol. Chem. 277, 44589-44592) that the folding of wild-type prion protein can best be described by a three-state sequential model involving a partially folded intermediate. Here we have performed kinetic stopped-flow studies for a number of recombinant prion protein variants carrying mutations associated with familial forms of prion disease. Analysis of kinetic data clearly demonstrates the presence of partially structured intermediates on the refolding pathway of each PrP variant studied. In each case, the partially folded state is at least one order of magnitude more populated than the fully unfolded state. The present study also reveals that, for the majority of PrP variants tested, mutations linked to familial prion diseases result in a pronounced increase in the thermodynamic stability, and thus the population, of the folding intermediate. These data strongly suggest that partially structured intermediates of PrP may play a crucial role in prion protein conversion, serving as direct precursors of the pathogenic PrP(Sc) isoform. PMID- 14761943 TI - The binding of C10 oligomers to Escherichia coli transcription termination factor Rho. AB - The binding of C10 RNA oligomers to wild type and mutant Escherichia coli transcription termination factor Rho provides a model for the enzyme-RNA interactions that lead to transcription termination. One surprising finding is that wild type Rho binds between five and six C10 oligomers per hexamer with KD = 0.3 microm, and five to six additional C10 molecules with KD = 7 microm. Previously, approximately half this number of oligomer-binding sites was reported (Wang, Y., and von Hippel, P. H. (1993) J. Biol. Chem. 268, 13947-13955); however, the E155K mutant form of Rho, thought at the time to be wild type, was used in that work. The present results with E155K Rho agree with the earlier work. C10 binding with mutant forms of Rho that are altered in RNA interactions, bearing amino acid changes F62S, G99V, F232C, T286A, or K352E, indicate that the higher affinity binding sites constitute what has been termed the primary RNA site, and the lower affinity sites constitute the secondary sites. The binding data together with the crystal structures for wild type Rho (Skordalakes, E., and Berger, J. M. (2003) Cell 114, 135-146) support structurally distinct locations on Rho for the two classes of C10-binding sites. The results are consistent with participation of residues 33 A apart in secondary site RNA interactions. The data further indicate that not all RNA sites on Rho must be filled for full ATPase and transcription termination activity, and suggest a model in which RNA binding to the higher affinity sites leads to a protein conformation change that exposes the previously hidden lower affinity sites. PMID- 14761944 TI - TSG101 interacts with apoptosis-antagonizing transcription factor and enhances androgen receptor-mediated transcription by promoting its monoubiquitination. AB - Apoptosis-antagonizing transcription factor (AATF), also termed Che-1, was identified as interacting protein of Dlk/ZIP kinase and RNA polymerase II, respectively. Che-1 has additionally been shown to bind Rb, thereby activating transcription factor E2F and promoting cell cycle progression. Moreover, AATF enhances steroid receptor-mediated transactivation in a hormone- and dose dependent manner (Leister, P., Burgdorf, S., and Scheidtmann, K. H., (2003) Signal Transduction 3, 18-25). These data suggest that AATF exerts its functions through interaction with different transcription factors. In search of novel interaction partners of AATF, we identified the tumor susceptibility gene product TSG101, which had also been recognized as a co-regulator of nuclear hormone receptors. Interestingly, TSG101 and AATF functioned as cooperative coactivators in androgen receptor-mediated transcription. Because TSG101 was also shown to play a role in regulation of ubiquitin conjugation, we asked whether its coactivating function might be linked to ubiquitination. Indeed, TSG101 enhanced monoubiquitination of the androgen receptor in a ligand-dependent manner, and this correlated with enhanced transactivating capacity. Furthermore, a dominant negative mutant of ubiquitin preventing polyubiquitination also stimulated androgen receptor-mediated transcription, which in this case could not be enhanced by TSG101. We propose that TSG101 activates androgen receptor-induced transcription by transient stabilization of the monoubiquitinated state, thus revealing a novel regulatory mechanism for nuclear receptors. PMID- 14761946 TI - Identification of binding sites in the nicotinic acetylcholine receptor for [3H]azietomidate, a photoactivatable general anesthetic. AB - To identify binding domains in a ligand-gated ion channel for etomidate, an intravenous general anesthetic, we photolabeled nicotinic acetylcholine receptor (nAChR)-rich membranes from Torpedo electric organ with a photoactivatable analog, [(3)H]azietomidate. Based upon the inhibition of binding of the noncompetitive antagonist [(3)H]phencyclidine, azietomidate and etomidate bind with 10-fold higher affinity to nAChRs in the desensitized state (IC(50) = 70 microm) than in the closed channel state. In addition, both drugs between 0.1 and 1 mm produced a concentration-dependent enhancement of [(3)H]ACh equilibrium binding affinity, but they inhibited binding at higher concentrations. UV irradiation resulted in preferential [(3)H]azietomidate photoincorporation into the nAChR alpha and delta subunits. Photolabeled amino acids in both subunits were identified in the ion channel domain and in the ACh binding sites by Edman degradation. Within the nAChR ion channel in the desensitized state, there was labeling of alphaGlu-262 and deltaGln-276 at the extracellular end and deltaSer 258 and deltaSer-262 toward the cytoplasmic end. Within the acetylcholine binding sites, [(3)H]azietomidate photolabeled alphaTyr-93, alphaTyr-190, and alphaTyr 198 in the site at the alpha-gamma interface and deltaAsp-59 (but not the homologous position, gammaGlu-57). Increasing [(3)H]azietomidate concentration from 1.8 to 150 microm increased the efficiency of incorporation into amino acids within the ion channel by 10-fold and in the ACh sites by 100-fold, consistent with higher affinity binding within the ion channel. The state dependence and subunit selectivity of [(3)H]azietomidate photolabeling are discussed in terms of the structures of the nAChR transmembrane and extracellular domains. PMID- 14761945 TI - Role of group V phospholipase A2 in zymosan-induced eicosanoid generation and vascular permeability revealed by targeted gene disruption. AB - Conclusions regarding the contribution of low molecular weight secretory phospholipase A2 (sPLA2) enzymes in eicosanoid generation have relied on data obtained from transfected cells or the use of inhibitors that fail to discriminate between individual members of the large family of mammalian sPLA2 enzymes. To elucidate the role of group V sPLA2, we used targeted gene disruption to generate mice lacking this enzyme. Zymosan-induced generation of leukotriene C4 and prostaglandin E2 was attenuated approximately 50% in peritoneal macrophages from group V sPLA2-null mice compared with macrophages from wild-type littermates. Furthermore, the early phase of plasma exudation in response to intraperitoneal injection of zymosan and the accompanying in vivo generation of cysteinyl leukotrienes were markedly attenuated in group V sPLA2-null mice compared with wild-type controls. These data provide clear evidence of a role for group V sPLA2 in regulating eicosanoid generation in response to an acute innate stimulus of the immune response both in vitro and in vivo, suggesting a role for this enzyme in innate immunity. PMID- 14761947 TI - Extracellular ATP-mediated signaling for survival in hyperoxia-induced oxidative stress. AB - Respiratory failure is a serious consequence of lung cell injury caused by treatment with high inhaled oxygen concentrations. Human lung microvascular endothelial cells (HLMVEC) are a principal target of hyperoxic injury (hyperoxia). Cell stress can cause release of ATP, and this extracellular nucleotide can activate purinoreceptors and mediate responses essential for survival. In this investigation, exposure of endothelial cells to an oxidative stress, hyperoxia, caused rapid but transient ATP release (20.03 +/- 2.00 nm/10(6) cells in 95% O(2) versus 0.08 +/- 0.01 nm/10(6) cells in 21% O2 at 30 min) into the extracellular milieu without a concomitant change in intracellular ATP. Endogenously produced extracellular ATP-enhanced mTOR-dependent uptake of glucose (3467 +/- 102 cpm/mg protein in 95% oxygen versus 2100 +/- 112 cpm/mg protein in control). Extracellular addition of ATP-activated important cell survival proteins like PI 3-kinase and extracellular-regulated kinase (ERK-1/2). These events were mediated primarily by P2Y receptors, specifically the P2Y2 and/or P2Y6 subclass of receptors. Extracellular ATP was required for the survival of HLMVEC in hyperoxia (55 +/- 10% surviving cells with extracellular ATP scavengers [apyrase + adenosine deaminase] versus 95 +/- 12% surviving cells without ATP scavengers at 4 d of hyperoxia). Incubation with ATP scavengers abolished ATP-dependent ERK phosphorylation stimulated by hyperoxia. Further, ERK activation also was found to be important for cell survival in hyperoxia, as treatment with PD98059 enhanced hyperoxia-mediated cell death. These findings demonstrate that ATP release and subsequent ATP-mediated signaling events are vital for survival of HLMVEC in hyperoxia. PMID- 14761948 TI - Viral evolution as a tool to improve the tetracycline-regulated gene expression system. AB - We present viral evolution as a novel and powerful method to optimize non-viral proteins. We used this approach to optimize the tetracycline (Tc)-regulated gene expression system (Tet system) for its function in mammalian cells. The components of the Tet system were incorporated in the human immunodeficiency virus (HIV)-1 virus such that viral replication is controlled by this regulatory system. Upon long term replication of this HIV-rtTA virus in human T cells, we obtained a virus variant with an enhanced replication potential resulting from an improved rtTA component of the introduced Tet system. We identified a single amino acid exchange, F86Y, which enhances the transcriptional activity and doxycycline (dox) sensitivity of rtTA. We generated a new rtTA variant that is 5 fold more active at high dox levels than the initial rtTA, and 25-fold more sensitive to dox, whereas the background activity in the absence of dox is not increased. This new rtTA variant will be very useful in biological applications that require a more sensitive or active Tet system. Our results demonstrate that the viral evolution strategy can be used to improve the activity of genes by making them an integral and essential part of the virus. PMID- 14761949 TI - Hijacking of the human alkyl-N-purine-DNA glycosylase by 3,N4-ethenocytosine, a lipid peroxidation-induced DNA adduct. AB - Lipid peroxidation generates aldehydes, which react with DNA bases, forming genotoxic exocyclic etheno(epsilon)-adducts. E-bases have been implicated in vinyl chloride-induced carcinogenesis, and increased levels of these DNA lesions formed by endogenous processes are found in human degenerative disorders. E adducts are repaired by the base excision repair pathway. Here, we report the efficient biological hijacking of the human alkyl-N-purine-DNA glycosylase (ANPG) by 3,N(4)-ethenocytosine (epsilonC) when present in DNA. Unlike the ethenopurines, ANPG does not excise, but binds to epsilonC when present in either double-stranded or single-stranded DNA. We developed a direct assay, based on the fluorescence quenching mechanism of molecular beacons, to measure a DNA glycosylase activity. Molecular beacons containing modified residues have been used to demonstrate that the epsilonC.ANPG interaction inhibits excision repair both in reconstituted systems and in cultured human cells. Furthermore, we show that the epsilonC.ANPG complex blocks primer extension by the Klenow fragment of DNA polymerase I. These results suggest that epsilonC could be more genotoxic than 1,N(6)-ethenoadenine (epsilonA) residues in vivo. The proposed model of ANPG mediated genotoxicity of epsilonC provides a new insight in the molecular basis of lipid peroxidation-induced cell death and genome instability in cancer. PMID- 14761950 TI - Casein kinase 1 delta phosphorylates tau and disrupts its binding to microtubules. AB - Tau hyperphosphorylation precedes neuritic lesion formation in Alzheimer's disease, suggesting it participates in the tau fibrillization reaction pathway. Candidate tau protein kinases include members of the casein kinase 1 (CK1) family of phosphotransferases, which are highly overexpressed in Alzheimer's disease brain and colocalize with neuritic and granulovacuolar lesions. Here we characterized the contribution of one CK1 isoform, Ckidelta, to the phosphorylation of tau at residues Ser202/Thr205 and Ser396/Ser404 in human embryonic kidney 293 cells using immunodetection and fluorescence microscopy. Treatment of cells with membrane permeable CK1 inhibitor 3-[(2,3,6 trimethoxyphenyl)methylidenyl]-indolin-2-one (IC261) lowered occupancy of Ser396/Ser404 phosphorylation sites by >70% at saturation, suggesting that endogenous CK1 was the major source of basal phosphorylation activity at these sites. Overexpression of Ckidelta increased CK1 enzyme activity and further raised tau phosphorylation at residues Ser202/Thr205 and Ser396/Ser404 in situ. Inhibitor IC261 reversed tau hyperphosphorylation induced by Ckidelta overexpression. Co-immunoprecipitation assays showed direct association of tau and Ckidelta in situ, consistent with tau being a Ckidelta substrate. Ckidelta overexpression also produced a decrease in the fraction of bulk tau bound to detergent-insoluble microtubules. These results suggest that Ckidelta phosphorylates tau at sites that modulate tau/microtubule binding, and that the expression pattern of Ckidelta in Alzheimer's disease is consistent with it playing an important role in tau aggregation. PMID- 14761951 TI - Hif1 is a component of yeast histone acetyltransferase B, a complex mainly localized in the nucleus. AB - Hat1 is the catalytic subunit of the only type B histone acetyltransferase known (HAT-B). The enzyme specifically acetylates lysine 12, and to a lesser extent lysine 5, of free, non-chromatin-bound histone H4. The complex is usually isolated with cytosolic fractions and is thought to be involved in chromatin assembly. The Saccharomyces cerevisiae HAT-B complex also contains Hat2, a protein stimulating Hat1 catalytic activity. We have now identified by two-hybrid experiments Hif1 as both a Hat1- and a histone H4-interacting protein. These interactions were dependent on HAT2, indicating a mediating role for Hat2. Biochemical fractionation and co-immunoprecipitation assays demonstrated that Hif1 is a component of a yeast heterotrimeric HAT-B complex, in which Hat2 bridges Hat1 and Hif1 proteins. In contrast to Hat2, this novel subunit does not appear to regulate Hat1 enzymatic activity. Nevertheless, similarly to Hat1, Hif1 influences telomeric silencing. In a localization analysis by immunofluorescence microscopy on yeast strains expressing tagged versions of Hat1, Hat2, and Hif1, we have found that all three HAT-B proteins are mainly localized in the nucleus. Thus, we propose that the distinction between A- and B-type enzymes should henceforth be based on their capacity to acetylate histones bound to nucleosomes and not on their location within the cell. Finally, by Western blotting assays, we have not detected differences in the in vivo acetylation of H4 lysine 12 (acK12H4) between wild-type and hat1Delta, hat2Delta, or hif1Delta mutant strains, suggesting that the level of HAT-B-dependent acK12H4 may be very low under normal growth conditions. PMID- 14761952 TI - A conformation-specific interhelical salt bridge in the K+ binding site of gastric H,K-ATPase. AB - Homology modeling of gastric H,K-ATPase based on the E2 model of sarcoplasmic reticulum Ca2+-ATPase (Toyoshima, C., and Nomura, H. (2002) Nature 392, 835-839) revealed the presence of a single high-affinity binding site for K+ and an E2 form-specific salt bridge between Glu820 (M6) and Lys791 (M5). In the E820Q mutant this salt bridge is no longer possible, and the head group of Lys791, together with a water molecule, fills the position of the K+ ion and apparently mimics the K+-filled cation binding pocket. This gives an explanation for the K+ independent ATPase activity and dephosphorylation step of the E820Q mutant (Swarts, H. G. P., Hermsen, H. P. H., Koenderink, J. B., Schuurmans Stekhoven, F. M. A. H., and De Pont, J. J. H. H. M. (1998) EMBO J. 17, 3029-3035) and, indirectly, for its E1 preference. The model is strongly supported by a series of reported mutagenesis studies on charged and polar amino acid residues in the membrane domain. To further test this model, Lys791 was mutated alone and in combination with other crucial residues. In the K791A mutant, the K+ affinity was markedly reduced without altering the E2 preference of the enzyme. The K791A mutation prevented, in contrast to the K791R mutation, the spontaneous dephosphorylation of the E820Q mutant as well as its conformational equilibrium change toward E1. This indicates that the salt bridge is essential for high affinity K+ binding and the E2 preference of H,K-ATPase. Moreover, its breakage (E820Q) can generate a K+-insensitive activity and an E1 preference. In addition, the study gives a molecular explanation for the electroneutrality of H,K-ATPases. PMID- 14761953 TI - Anti-cooperative oxidation of ubiquinol by the yeast cytochrome bc1 complex. AB - We have investigated the interaction between monomers of the dimeric yeast cytochrome bc(1) complex by analyzing the pre-steady and steady state activities of the isolated enzyme in the presence of antimycin under conditions that allow the first turnover of ubiquinol oxidation to be observable in cytochrome c(1) reduction. At pH 8.8, where the redox potential of the iron-sulfur protein is approximately 200 mV and in a bc(1) complex with a mutated iron-sulfur protein of equally low redox potential, the amount of cytochrome c(1) reduced by several equivalents of decyl-ubiquinol in the presence of antimycin corresponded to only half of that present in the bc(1) complex. Similar experiments in the presence of several equivalents of cytochrome c also showed only half of the bc(1) complex participating in quinol oxidation. The extent of cytochrome b reduced corresponded to two b(H) hemes undergoing reduction through one center P per dimer, indicating electron transfer between the two cytochrome b subunits. Antimycin stimulated the ubiquinol-cytochrome c reductase activity of the bc(1) complex at low inhibitor/enzyme ratios. This stimulation could only be fitted to a model in which half of the bc(1) dimer is inactive when both center N sites are free, becoming active upon binding of one center N inhibitor molecule per dimer, and there is electron transfer between the cytochrome b subunits of the dimer. These results are consistent with an alternating half-of-the-sites mechanism of ubiquinol oxidation in the bc(1) complex dimer. PMID- 14761954 TI - Regulation of the type 1 inositol 1,4,5-trisphosphate receptor by phosphorylation at tyrosine 353. AB - The inositol 1,4,5-trisphosphate receptor (IP3R) plays an essential role in Ca2+ signaling during lymphocyte activation. Engagement of the T cell or B cell receptor by antigen initiates a signal transduction cascade that leads to tyrosine phosphorylation of IP3R by Src family nonreceptor protein tyrosine kinases, including Fyn. However, the effect of tyrosine phosphorylation on the IP3R and subsequent Ca2+ release is poorly understood. We have identified tyrosine 353 (Tyr353) in the IP3-binding domain of type 1 IP3R (IP3R1) as a phosphorylation site for Fyn both in vitro and in vivo. We have developed a phosphoepitope-specific antibody and shown that IP3R1-Y353 becomes phosphorylated during T cell and B cell activation. Furthermore, tyrosine phosphorylation of IP3R1 increased IP3 binding at low IP3 concentrations (<10 nm). Using wild-type IP3R1 or an IP3R1-Y353F mutant that cannot be tyrosine phosphorylated at Tyr353 or expressed in IP3R-deficient DT40 B cells, we demonstrated that tyrosine phosphorylation of Tyr353 permits prolonged intracellular Ca2+ release during B cell activation. Taken together, these data suggest that one function of tyrosine phosphorylation of IP3R1-Y353 is to enhance Ca2+ signaling in lymphocytes by increasing the sensitivity of IP3R1 to activation by low levels of IP3. PMID- 14761955 TI - Human Sgt1 binds HSP90 through the CHORD-Sgt1 domain and not the tetratricopeptide repeat domain. AB - Sgt1 has been identified as a subunit of both core kinetochore and SCF (Skp1-Cul1 F-box) ubiquitin ligase complexes and is also implicated in plant disease resistance. Sgt1 has two putative HSP90 binding domains, a tetratricopeptide repeat and a p23-like CHORD and Sgt1 (CS) domain. Using NMR spectroscopy, we show that only the CS domain of human Sgt1 physically interacts with HSP90. The tetratricopeptide repeat domain does not bind to either HSP90 or HSP70. Determination of the three-dimensional structure showed that the Sgt1-CS domain shares the same beta-sandwich fold as p23 but lacks the last highly conserved beta-strand in p23. Analysis of the structures of Sgt1-CS and p23 revealed a similar charge distribution on one of two opposing surfaces that suggests that it is the binding region for HSP90 in Sgt1. Although ATP is absolutely required for p23 binding to HSP90, Sgt1 binds to HSP90 also in the absence of the non hydrolyzable analog ATPgammaS. Our findings suggest the CS domain is a binding module for HSP90 distinct from p23-like domains, which implies that Sgt1 and related proteins function in recruiting heat shock protein activities to multiprotein assemblies. PMID- 14761956 TI - Ectodomain shedding of the neural recognition molecule CHL1 by the metalloprotease-disintegrin ADAM8 promotes neurite outgrowth and suppresses neuronal cell death. AB - The neural cell adhesion molecule "close homologue of L1," termed CHL1, has functional importance in the nervous system. CHL1 is expressed as a transmembrane protein of 185 kDa, and ectodomain shedding releases soluble fragments relevant for its physiological function. Here we describe that ADAM8, a member of the family of metalloprotease disintegrins cleaves a CHL1-Fc fusion protein in vitro at two sites corresponding to release of the extracellular domain of CHL1 in fibronectin (FN) domains II (125 kDa) and V (165 kDa), inhibited by batimastat (BB-94). Cleavage of CHL1-Fc in the 125-kDa fragment was not detectable under non reducing conditions arguing that cleavage resulting in the 165-kDa fragment is more relevant in releasing soluble CHL1 in vivo. In cells transfected with full length ADAM8, membrane proximal cleavage of CHL1 was similar and not stimulated by phorbol ester 12-O-tetradecanoylphorbol-13-acetate and pervanadate. No cleavage of CHL1 was observed in cells expressing either inactive ADAM8 with a Glu330 to Gln exchange (EQ-A8), or active ADAM10 and ADAM17. Consequently, processing of CHL1 was hardly detectable in brain extracts of ADAM8-deficient mice but enhanced in a neurodegenerative mouse mutant. CHL1 processed by ADAM8 in supernatants of COS-7 cells and in co-culture with cerebellar granule neurons was very potent in stimulating neurite outgrowth and suppressing neuronal cell death, not observed in cells co-transfected with CHL1/EQ-A8, CHL1/ADAM10, or CHL1/ADAM17. Taken together, we propose that ADAM8 plays an important role in physiological and pathological cell interactions by a specific release of functional CHL1 from the cell surface. PMID- 14761957 TI - Sodium channel beta1 subunit-mediated modulation of Nav1.2 currents and cell surface density is dependent on interactions with contactin and ankyrin. AB - Voltage-gated sodium channels are composed of a pore-forming alpha subunit and at least one auxiliary beta subunit. Both beta1 and beta2 are cell adhesion molecules that interact homophilically, resulting in ankyrin recruitment. In contrast, beta1, but not beta2, interacts heterophilically with contactin, resulting in increased levels of cell surface sodium channels. We took advantage of these results to investigate the molecular basis of beta1-mediated enhancement of sodium channel cell surface density, including elucidating structure-function relationships for beta1 association with contactin, ankyrin, and Nav1.2. beta1/beta2 subunit chimeras were used to assign putative sites of contactin interaction to two regions of the beta1 Ig loop. Recent studies have shown that glutathione S-transferase fusion proteins containing portions of Nav1.2 intracellular domains interact directly with ankyrinG. We show that native Nav1.2 associates with ankyrinG in cells in the absence of beta subunits and that this interaction is enhanced in the presence of beta1 but not beta1Y181E, a mutant that does not interact with ankyrinG. beta1Y181E does not modulate Nav1.2 channel function despite efficient association with Nav1.2 and contactin. beta1Y181E increases Nav1.2 cell surface expression, but not as efficiently as wild type beta1. beta1/beta2 chimeras exchanging various regions of the beta1 Ig loop were all ineffective in increasing Nav1.2 cell surface density. Our results demonstrate that full-length beta1 is required for channel modulation and enhancement of sodium channel cell surface expression. PMID- 14761958 TI - State-specific monoclonal antibodies identify an intermediate state in epsilon protein kinase C activation. AB - Evaluation of the activation state of protein kinase C (PKC) isozymes relies on analysis of subcellular translocation. A monoclonal antibody, 14E6, specific for the activated conformation of epsilonPKC, was raised using the first variable (V1) domain of epsilonPKC as the immunogen. 14E6 binding is specific for epsilonPKC and is greatly increased in the presence of PKC activators. Immunofluorescence staining by 14E6 of neonatal rat primary cardiac myocytes and the NG108-15 neuroblastoma glioma cell line, NG108-15/D2, increases rapidly following cell activation and is localized to new subcellular sites. However, staining of translocated epsilonPKC with 14E6 is transient, and the epitope disappears 30 min after activation of NG-108/15 cells by a D2 receptor agonist. In contrast, subcellular localization associated with activation, as determined by commercially available polyclonal antibodies, persists for at least 30 min. In vitro, epsilonRACK, the receptor for activated epsilonPKC, inhibits 14E6 binding to epsilonPKC, suggesting that the 14E6 epitope is lost or hidden when active epsilonPKC binds to its RACK. Therefore, the 14E6 antibody appears to identify a transient state of activated but non-anchored epsilonPKC. Moreover, binding of 14E6 to epsilonPKC only after activation suggests that lipid-dependent conformational changes associated with epsilonPKC activation precede binding of the activated isozyme to its specific RACK, epsilonRACK. Further, monoclonal antibody 14E6 should be a powerful tool to study the pathways that control rapid translocation of epsilonPKC from cytosolic to membrane localization on activation. PMID- 14761959 TI - HFA1 encoding an organelle-specific acetyl-CoA carboxylase controls mitochondrial fatty acid synthesis in Saccharomyces cerevisiae. AB - The Saccharomyces cerevisiae gene, HFA1, encodes a >250-kDa protein, which is required for mitochondrial function. Hfa1p exhibits 72% overall sequence similarity (54% identity) to ACC1-encoded yeast cytoplasmic acetyl-CoA carboxylase. Nevertheless, HFA1 and ACC1 functions are not overlapping because mutants of the two genes have different phenotypes and do not complement each other. Whereas ACC1 is involved in cytoplasmic fatty acid synthesis, the phenotype of hfa1Delta disruptants resembles that of mitochondrial fatty-acid synthase mutants. They fail to grow on lactate or glycerol, and the mitochondrial cofactor, lipoic acid, is reduced to <10% of its normal cellular concentration. Other than Acc1p, the N-terminal sequence of Hfa1p comprises a canonical mitochondrial targeting signal together with a matrix protease cleavage site. Accordingly, the HFA1-encoded protein was specifically assigned by Western blotting of appropriate cell fractions to the mitochondrial compartment. Removal of the mitochondrial targeting sequence abolished the competence of HFA1 DNA to complement hfal null mutants. Conversely and in contrast to the intact HFA1 sequence, the signal sequence-free HFA1 gene complemented the mutational loss of cytoplasmic acetyl-CoA carboxylase. Expression of HFA1 under the control of the ACC1 promoter restored cellular ACC activity in ACC1-defective yeast mutants to wild type levels. From this finding, it is concluded that HFA1 encodes a specific mitochondrial acetyl-CoA carboxylase providing malonyl-CoA for intraorganellar fatty acid and, in particular, lipoic acid synthesis. PMID- 14761960 TI - Interaction of estrogen receptor alpha with 3-methyladenine DNA glycosylase modulates transcription and DNA repair. AB - Estrogen receptor alpha (ERalpha) interacts with basal transcription factors, coregulatory proteins, and chromatin modifiers to initiate transcription of the target genes. We have identified a novel interaction between ERalpha and the DNA repair protein 3-methyladenine DNA glycosylase (MPG) thereby providing a functional link between gene expression and DNA repair. Interestingly, the ERalpha-MPG interaction was enhanced by the presence of estrogen response element (ERE)-containing DNA. In vitro pull-down assays indicated that the interaction of ERalpha with MPG was direct and occurred through the DNA- and ligand-binding domains and the hinge region of the receptor. More importantly, endogenously expressed ERalpha and MPG from MCF-7 cells coimmunoprecipitated with ERalpha- and MPG-specific antibodies. The ERalpha-MPG interaction had functional consequences on the activities of both proteins. ERalpha increased MPG acetylation, stabilized the binding of MPG with hypoxanthine-containing oligos, and enhanced MPG catalyzed removal of hypoxanthine from DNA. In turn, MPG dramatically stabilized the interaction of ERalpha with ERE-containing oligos, decreased p300-mediated acetylation of the receptor, and reduced transcription of simple and complex ERE containing reporter plasmids in a dose-dependent manner. Our studies suggest that recruitment of MPG to ERE-containing genes influences transcription and plays a role in maintaining integrity of the genome by recruiting DNA repair proteins to actively transcribing DNA. PMID- 14761961 TI - Domain analysis of the calcium-activated potassium channel SK1 from rat brain. Functional expression and toxin sensitivity. AB - Two small conductance, calcium-activated potassium channels (SK channels), SK2 and SK3, have been shown to contribute to the afterhyperpolarization (AHP) and to shape the firing behavior in neurons for example in the hippocampal formation, the dorsal vagal nucleus, the subthalamic nucleus, and the cerebellum. In heterologous expression systems, SK2 and SK3 currents are blocked by the bee venom toxin apamin, just as well as the corresponding neuronal AHP currents. However, the functional role and pharmacological profile of SK1 channels from rat brain (rSK1) is still largely unknown, as so far rSK1 homomeric channels could not be functionally expressed. We have performed a domain analysis to elucidate the pharmacological profile and the molecular determinants of rSK1 channel expression by using channel chimeras in combination with immunocytochemistry, immunoblot analysis, and electrophysiology. Our results reveal that the rSK1 subunit is synthesized in cells but does not form functional homomeric channels. Exchanging the carboxyl terminus of rSK1 for that of hSK1 or rSK2 is sufficient to rescue the functional expression of rSK1 channels. Additionally, transplantation of both amino and carboxyl termini of rSK1 onto hSK1 subunits, normally forming functional homomeric channel, hinders their functional expression, while hSK1 channels containing only the rSK1 carboxyl terminus are functional. These results suggest that the lack of functional expression of rSK1 channels is probably due to problems in their assembly and tetramerization but not in their calmodulin-dependent gating. Finally, we show that chimeric channels containing the core domain (S1-S6) of rSK1, unlike hSK1, are apamin-insensitive. PMID- 14761962 TI - NAD(P)H oxidase-derived H(2)O(2) signals chloride channel activation in cell volume regulation and cell proliferation. AB - Cellular swelling triggers the activation of Cl(-) channels (volume-sensitive outwardly rectifying (VSOR) Cl(-) channels) in many cell types. Ensuing regulatory volume decrease has been considered the primary function of these channels. However, Cl(-) channels, which share functional properties with volume sensitive Cl(-) channels, have been shown to be involved in other physiological processes, including cell proliferation and apoptosis, raising the question of their physiological roles and the signal transduction pathways involved in their activation. Here we report that exogenously applied H(2)O(2) elicited VSOR Cl(-) channel activation. Furthermore, activation of these channels was found to be coupled to NAD(P)H oxidase activity. Also, epidermal growth factor, known to increase H(2)O(2) production, activated Cl(-) channels with properties identical to swelling-sensitive Cl(-) channels. It is concluded that NAD(P)H oxidase derived H(2)O(2) is the common signal transducing molecule that mediates the activation of these ubiquitously expressed anion channels under a variety of physiological conditions. PMID- 14761963 TI - Positive regulation of apoptosis signal-regulating kinase 1 by hD53L1. AB - Apoptosis signal-regulating kinase 1 (ASK1) is a mitogen-activated protein kinase kinase kinase family member that plays a central role in cytokine- and stress induced apoptosis by activating c-Jun N-terminal kinase and p38 signaling cascades. ASK1-induced apoptotic activity is up-regulated by two cellular factors, Daxx and TRAF2, through direct protein-protein interactions. Daxx and TRAF2 are death receptor-associated proteins in Fas and tumor necrosis factor alpha pathways, respectively. Recent studies suggest that calcium signaling may regulate ASK1 pathway. Here we report that human D53L1, a member of the tumor protein D52 family involved in cell proliferation and calcium signaling, up regulates the ASK1-induced apoptosis. The human D53L1 physically interacts with the C-terminal regulatory domain of ASK1 and promotes ASK1-induced apoptotic activity by activating caspase signaling in mammalian cells. In luciferase reporter assays, hD53L1 activates c-Jun N-terminal kinase-mediated transactivation in the presence of ASK1. Expression of hD53L1 enhances autophosphorylation and kinase activity of ASK1 but has no effect on ASK1 oligomerization that is necessary for kinase activity and on binding of ASK1 to MKK6, a downstream factor of ASK1. Taken together, these results suggest that activation of ASK1 by hD53L1 may provide a novel mechanism for ASK1 regulation. PMID- 14761964 TI - Characterization of proteins binding to E-box/Ku86 sites and function of Ku86 in transcriptional regulation of the human xanthine oxidoreductase gene. AB - We reported previously that E-box and TATA-like elements repress human xanthine oxidoreductase gene (hXOR) expression. In the present investigation, we determined the means by which the E-box site functions in this basal repression. DNA affinity purification demonstrated that at least five proteins are involved in the nuclear protein complex binding to the E-box and adjacent Ku86-binding sites. Amino acid sequence analysis demonstrated that three proteins, DNA-PK catalytic subunit, Ku86, and Ku70 are components of DNA-dependent protein kinase (DNA-PK). By electrophoretic mobility shift assays, gel-shift, and site-directed mutagenesis, we confirmed Ku86 binding to the Ku86 site. Studies indicated that the other two proteins of the complex are AREB6-like proteins binding to the E box. Pull-down and immunoprecipitation analyses demonstrated the binding of Ku86 to AREB6-like proteins. The functional loss of Ku86 increases hXOR promoter activity and transcript expression. Based on the findings, we propose that DNA PK/AREB6-like proteins play a central role in repression of basal hXOR activity. AREB6-like proteins specifically bind to the E-box, whereas Ku86 binds an adjacent site and recruits DNA-PK catalytic subunit and Ku70 proteins. A working model is presented to account for the role of DNA-PK and AREB6-like proteins in regulating hXOR activity. PMID- 14761965 TI - Regulation of a gamma-aminobutyric acid transporter by reciprocal tyrosine and serine phosphorylation. AB - A feature of the rat brain gamma-aminobutyric acid transporter GAT1, and other members of the neurotransmitter transporter family, is its regulated redistribution between intracellular locations and the plasma membrane. Recent studies have focused upon defining the signaling molecules that facilitate this redistribution. Agents that promote direct tyrosine phosphorylation of GAT1 promote a relative increase in surface GAT1 levels, and this results from a slowing of the transporter internalization rate. Agents that act to increase protein kinase C (PKC) activity promote a relative decrease in surface GAT1 levels; whether this effect is caused by direct transporter phosphorylation is unknown. The opposing actions of tyrosine kinase activity and PKC activity raise the possibility that the subcellular distribution of GAT1 is associated with mutually exclusive transporter phosphorylation events. The present experiments show that GAT1 is phosphorylated on serine residues in a PKC-dependent manner, but this state is only revealed when GAT1 tyrosine phosphorylation is eliminated or greatly reduced. The relative levels of serine phosphorylation and tyrosine phosphorylation are negatively correlated. The amount of serine phosphorylation is regulated by agents that affect tyrosine phosphorylation, and vice versa. In addition, the ability of agents that affect tyrosine kinase activity to regulate GAT1 serine phosphorylation requires a change in its tyrosine phosphorylation state. These data support the ideas that GAT1 can exist in either of two mutually exclusive phosphorylation states and that the relative abundance of these states determines in part the relative subcellular distribution of the transporter. PMID- 14761966 TI - Formation of protein tyrosine ortho-semiquinone radical and nitrotyrosine from cytochrome c-derived tyrosyl radical. AB - Oxidative alteration of mitochondrial cytochrome c (cyt c) has been linked to disease pathophysiology and is one of the causative factors for pro-apoptotic events. Hydrogen peroxide induces a short-lived cyt c-derived tyrosyl radical as detected by the electron spin resonance (ESR) spin-trapping technique. This investigation was undertaken to characterize the fate and consequences of the cyt c-derived tyrosyl radical. The direct ESR spectrum from the reaction of cyt c with H(2)O(2) revealed a single-line signal with a line width of approximately 10 G. The detected ESR signal could be prevented by pretreatment of cyt c with iodination, implying that the tyrosine residue of cyt c was involved. The ESR signal can be enhanced and stabilized by a divalent metal ion such as Zn(2+), indicating the formation of the protein tyrosine ortho-semiquinone radical (ToQ.). The production of cyt c-derived ToQ. is inhibited by the spin trap, 2 methyl-2-nitrosopropane (MNP), suggesting the participation of tyrosyl radical in the formation of the ortho-semiquinone radical. The endothelium relaxant factor nitric oxide is well known to mediate mitochondrial respiration and apoptosis. The consumption of NO by cyt c was enhanced by addition of H(2)O(2) as verified by inhibition electrochemical detection using an NO electrode. The rate of NO consumption in the system containing cyt c/NO/H(2)O(2) was decreased by the spin traps 5,5-dimethyl pyrroline N-oxide and MNP, suggesting NO trapping of the cyt c derived tyrosyl radical. The above result was further confirmed by NO quenching of the ESR signal of the MNP adduct of cyt c tyrosyl radical. Immunoblotting analysis of cyt c after exposure to NO in the presence of H(2)O(2) revealed the formation of 3-nitrotyrosine. The addition of superoxide dismutase did not change the cyt c nitration, indicating that it is peroxynitrite-independent. The results of this study may provide useful information in understanding the interconnection among cyt c, H(2)O(2), NO, and apoptosis. PMID- 14761967 TI - Docking of endothelial nitric oxide synthase (eNOS) to the mitochondrial outer membrane: a pentabasic amino acid sequence in the autoinhibitory domain of eNOS targets a proteinase K-cleavable peptide on the cytoplasmic face of mitochondria. AB - Despite growing evidence for a mitochondrial localization of nitric oxide (NO) synthase and a broadening spectrum of NO actions on mitochondrial respiration and apoptosis, the basis for interaction between the enzyme and the organelle remain obscure. Here we investigated mitochondrial localization of endothelial nitric oxide synthase (eNOS) in human umbilical vein endothelial cells and human embryonic kidney cells transfected or infected with eNOS expression vectors. Copurification of eNOS with mitochondria was observed in both human umbilical vein endothelial cells and eNOS-expressing human embryonic kidney cells. Immunodetectable eNOS was cleaved from mitochondria by proteinase K treatment, suggesting eNOS association with the outer mitochondrial membrane. Localization of eNOS to a proteinase K-cleavable site on the cytoplasmic face of the outer membrane was confirmed by immunogold labeling of non-permeabilized mitochondria. Markers for mitochondrial subfractions ruled out the possibility of eNOS association with an intramitochondrial site or inverted mitochondrial particles. Denaturation of eNOS did not attenuate association with mitochondria. Mutant eNOS lacking a pentabasic amino acid sequence within the autoinhibitory domain (residues 628-632 of the bovine eNOS) showed dramatically reduced binding to the mitochondrial but not to the plasma membrane, which was associated with increased oxygen consumption. Collectively, these findings argue in favor of eNOS localization to the outer mitochondrial membrane in endothelial cells and identify elements of a novel anchoring mechanism. PMID- 14761968 TI - NH3 is involved in the NH4+ transport induced by the functional expression of the human Rh C glycoprotein. AB - Renal ammonium (NH3 + NH4+) transport is a key process for body acid-base balance. It is well known that several ionic transport systems allow NH4+ transmembrane translocation without high specificity NH4+, but it is still debated whether NH3, and more generally, gas, may be transported by transmembrane proteins. The human Rh glycoproteins have been proposed to mediate ammonium transport. Transport of NH4+ and/or NH3 by the epithelial Rh C glycoprotein (RhCG) may be of physiological importance in renal ammonium excretion because RhCG is mainly expressed in the distal nephron. However, RhCG function is not yet established. In the present study, we search for ammonium transport by RhCG. RhCG function was investigated by electrophysiological approaches in RhCG-expressing Xenopus laevis oocytes. In the submillimolar concentration range, NH4Cl exposure induced inward currents (IAM) in voltage-clamped RhCG-expressing cells, but not in control cells. At physiological extracellular pH (pHo) = 7.5, the amplitude of IAM increased with NH4Cl concentration and membrane hyperpolarization. The amplitude of IAM was independent of external Na+ or K+ concentrations but was enhanced by alkaline pHo and decreased by acid pHo. The apparent affinity of RhCG for NH4+ was affected by NH3 concentration and by changing pHo, whereas the apparent affinity for NH3 was unchanged by pHo, consistent with direct NH3 involvement in RhCG function. The enhancement of methylammonium-induced current by NH3 further supported this conclusion. Exposure to 500 microm NH4Cl induced a biphasic intracellular pH change in RhCG-expressing oocytes, consistent with both NH3 and NH4+ enhanced influx. Our results support the hypothesis of a specific role for RhCG in NH3 and NH4+ transport. PMID- 14761969 TI - Epigenetic control of mouse Oct-4 gene expression in embryonic stem cells and trophoblast stem cells. AB - The first cell differentiation event in mammalian embryogenesis segregates inner cell mass lineage from the trophectoderm at the blastocyst stage. Oct-4, a member of the POU family of transcription factors, is necessary for the pluripotency of the inner cell mass lineage. Embryonic stem (ES) cells, which contribute to all of embryonic lineages, express the Oct-4 gene. Trophoblast stem (TS) cells, which have the ability to differentiate into trophoblast lineage in vitro, never contribute to embryonic proper tissues in chimeras and differentiate only into trophoblastic cells in the placenta. Expression of the Oct-4 gene was undetectable and severely repressed in trophoblastic lineage, including the stem cells. We found that the culture of TS cells with 5-aza-2'-deoxycytidine or trichostatin A caused the activation of the Oct-4 gene. Analysis of the DNA methylation status of mouse Oct-4 gene upstream region revealed that Oct-4 enhancer/promoter region was hypomethylated in ES cells but hypermethylated in TS cells. Furthermore, in vitro methylation suppressed Oct-4 enhancer/promoter activity in reporter assay. In the placenta of Dnmt1(n/n) mutant mice, most of the CpGs in the enhancer/promoter region were unmethylated, and Oct-4 gene expression was aberrantly detected. Chromatin immunoprecipitation assay revealed that Oct-4 enhancer/promoter region was hyperacetylated in ES cells compared with TS cells, thus demonstrating that DNA methylation status is closely linked to the chromatin structure of the Oct-4 gene. Here we propose that the epigenetic mechanism, consisting of DNA methylation and chromatin remodeling, underlies the developmental stage- and cell type-specific mechanism of Oct-4 gene expression. PMID- 14761970 TI - The Id2 transcriptional repressor is induced by follicle-stimulating hormone and cAMP. AB - Id (inhibitor of DNA binding/differentiation) proteins repress differentiation and promote cell division by dimerizing with and inhibiting the action of basic helix-loop-helix transcription factors including those that bind to E-box motifs. Of the four characterized Id proteins, only Id2 is found in the nucleus of Sertoli cells that support the development of spermatozoa in the testis. Differential display analysis of rat primary Sertoli cell mRNA identified Id2 as being inducible by forskolin, a stimulator of cAMP production. Northern blot analysis confirmed that Id2 mRNA expression peaked in Sertoli cells 6-12 h after stimulation with forskolin or follicle-stimulating hormone (FSH), the major physiological stimulator of cAMP in Sertoli cells. Similarly, Id2 promoter activity in Sertoli cells was induced after forskolin or FSH stimulation as well as by overexpression of protein kinase A. Forskolin induction of the Id2 promoter required sequences located between positions -122 and -82. Protein(s) of 40-45 kDa were found to bind two activated transcription factor/cAMP-response element like sites and a GATA motif within the regulatory region. The induction of the Id2 gene by FSH corresponded with a decrease in protein binding to an E-box consensus motif and decreased E-box-mediated transcription. Together, these findings raise the possibility that FSH-mediated induction of Id2 and resultant inhibition of basic helix-loop-helix transcription factor-regulated genes in Sertoli cells may contribute to the regulation of spermatogenesis. PMID- 14761971 TI - CELF6, a member of the CELF family of RNA-binding proteins, regulates muscle specific splicing enhancer-dependent alternative splicing. AB - We previously described a family of five RNA-binding proteins: CUG-binding protein, embryonic lethal abnormal vision-type RNA-binding protein 3, and the CUG binding protein and embryonic lethal abnormal vision-type RNA-binding protein 3 like factors (CELFs) 3, 4, and 5. We demonstrated that all five of these proteins specifically activate exon inclusion of cardiac troponin T minigenes in vivo via muscle-specific splicing enhancer (MSE) sequences. We also predicted that a sixth family member, CELF6, was located on chromosome 15. Here, we describe the isolation and characterization of CELF6. Like the previously described CELF proteins, CELF6 shares a domain structure containing three RNA-binding domains and a divergent domain of unknown function. CELF6 is strongly expressed in kidney, brain, and testis and is expressed at very low levels in most other tissues. In the brain, expression is widespread and maintained from the fetus to the adult. CELF6 activates exon inclusion of a cardiac troponin T minigene in transient transfection assays in an MSE-dependent manner and can activate inclusion via multiple copies of a single element, MSE2. These results place CELF6 in a functional subfamily of CELF proteins that includes CELFs 3, 4, and 5. CELF6 also promotes skipping of exon 11 of insulin receptor, a known target of CELF activity that is expressed in kidney. PMID- 14761972 TI - Regulation of TRPC6 channel activity by tyrosine phosphorylation. AB - Various hormonal stimuli and growth factors activate the mammalian canonical transient receptor potential (TRPC) channel through phospholipase C (PLC) activation. However, the precise mechanism of the regulation of TRPC channel activity remains unknown. Here, we provide the first evidence that direct tyrosine phosphorylation by Src family protein-tyrosine kinases (PTKs) is a novel mechanism for modulating TRPC6 channel activity. We found that TRPC6 is tyrosine phosphorylated in COS-7 cells when coexpressed with Fyn, a member of the Src family PTKs. We also found that Fyn interacts with TRPC6 and that the interaction is mediated by the SH2 domain of Fyn and the N-terminal region of TRPC6 in a phosphorylation-independent manner. In addition, we demonstrated the physical association of TRPC6 with Fyn in the mammalian brain. Moreover, we showed that stimulation of the epidermal growth factor receptor induced rapid tyrosine phosphorylation of TRPC6 in COS-7 cells. This epidermal growth factor-induced tyrosine phosphorylation of TRPC6 was significantly blocked by PP2, a specific inhibitor of Src family PTKs, and by a dominant negative form of Fyn, suggesting that the direct phosphorylation of TRPC6 by Src family PTKs could be caused by physiological stimulation. Furthermore, using single channel recording, we showed that Fyn modulates TRPC6 channel activity via tyrosine phosphorylation. Thus, our findings demonstrated that tyrosine phosphorylation by Src family PTKs is a novel regulatory mechanism of TRPC6 channel activity. PMID- 14761973 TI - Functional molecular mapping of archaeal translation initiation factor 2. AB - Eukaryotic and archaeal initiation factors 2 (e/aIF2) are heterotrimeric proteins (alphabetagamma) carrying methionylated initiator tRNA to the small subunit of the ribosome. The three-dimensional structure of aIF2gamma from the Archaea Pyrococcus abyssi was previously solved. This subunit forms the core of the heterotrimer. The alpha and beta subunits bind the gamma, but do not interact together. aIF2gamma shows a high resemblance with elongation factor EF1-A. In this study, we characterize the role of each subunit in the binding of the methionylated initiator tRNA. Studying various aminoacyl-tRNA ligands shows that the methionyl group is a major determinant for recognition by aIF2. aIF2gamma alone is able to specifically bind Met-tRNAiMet, although with a reduced affinity as compared with the intact trimer. Site-directed mutagenesis confirms a binding mode of the tRNA molecule similar to that observed with the elongation factor. Under our assay conditions, aIF2beta is not involved in the docking of the tRNA molecule. In contrast, aIF2alpha provides the heterotrimer its full tRNA binding affinity. Furthermore, the isolated C-domain of aIF2alpha is responsible for binding of the alpha subunit to gamma. This binding involves an idiosyncratic loop of domain 2 of aIF2gamma. Association of the C-domain of aIF2alpha to aIF2gamma is enough to retrieve the binding affinity of tRNA for aIF2. The N terminal and central domains of aIF2alpha do not interfere with tRNA binding. However, the N-domain of aIF2alpha interacts with RNA unspecifically. Based on this property, a possible contribution of aIF2alpha to formation of a productive complex between aIF2 and the small ribosomal subunit is envisaged. PMID- 14761974 TI - Crystal structures of glutamine:phenylpyruvate aminotransferase from Thermus thermophilus HB8: induced fit and substrate recognition. AB - The following three-dimensional structures of three forms of glutamine:phenylpyruvate aminotransferase from Thermus thermophilus HB8 have been determined and represent the first x-ray analysis of the enzyme: the unliganded pyridoxal 5'-phosphate form at 1.9 A resolution and two complexes with 3 phenylpropionate and alpha-keto-gamma-methylthiobutyrate at 2.35 and 2.6 A resolution, respectively. The enzyme shows high activity toward phenylalanine, tyrosine, tryptophan, kynurenine, methionine, and glutamine. The enzyme is a homodimer, and each subunit is divided into an N-terminal arm and small and large domains. Based on its folding, the enzyme belongs to fold type I, aminotransferase subclass Ib. The subclass I aminotransferases whose structures have so far been determined exhibit a large movement of the small domain region upon binding of a substrate. Similarly, the glutamine:phenylpyruvate aminotransferase undergoes a large movement in part of the small domain to close the active site. The active-site pocket has a shape and size suitable to enclose the side chain of an aromatic amino acid or that of methionine. The inner side of the pocket is mostly hydrophobic, but also has polar sites. The kynurenine complex generated by computer modeling fits the pocket of the enzyme and its hydrophilic groups interact with the polar sites of the pocket. PMID- 14761975 TI - The tetrahydropyranopterin structure of the sulfur-free and metal-free molybdenum cofactor precursor. AB - The molybdenum cofactor (Moco), a highly conserved pterin compound coordinating molybdenum (Mo), is required for the activity of all Mo-dependent enzymes with the exception of nitrogenase. Moco is synthesized by a unique and evolutionary old multi-step pathway with two intermediates identified so far, the sulfur-free and metal-free pterin derivative precursor Z and molybdopterin, a pterin with an enedithiolate function essential for Mo ligation. The latter pterin component is believed to form a tetrahydropyranopterin similar to the one found for Moco in the crystal structure of Mo as well as tungsten (W) enzymes. Here we report the spectroscopic characterization and structure elucidation of precursor Z purified from Escherichia coli overproducing MoaA and MoaC, two proteins essential for bacterial precursor Z synthesis. We have shown that purified precursor Z is as active as precursor Z present in E. coli cell extracts, demonstrating that no modifications during the purification procedure have occurred. High resolution electrospray ionization mass spectrometry afforded a [M + H]+ ion compatible with a molecular formula of C10H15N5O8P. Consequently 1H NMR spectroscopy not allowed structural characterization of the molecule but confirmed that this intermediate undergoes direct oxidation to the previously well characterized non-productive follow-up product compound Z. The 1H chemical shift and coupling constant data are incompatible with previous structural proposals and indicate that precursor Z already is a tetrahydropyranopterin system and carries a geminal diol function in the C1' position. PMID- 14761976 TI - PIKE (phosphatidylinositol 3-kinase enhancer)-A GTPase stimulates Akt activity and mediates cellular invasion. AB - Akt/PKB is a crucial regulator of diverse cellular processes and contributes to cancer progression. Activation of Akt is essentially dependent on phosphatidylinositol (PI) 3-kinase signaling. Here, we describe a novel mediator of Akt that is independent of PI 3-kinase. This mediator, PIKE-A, is a PIKE isoform and contains GTPase, pleckstrin homology, ArfGAP, and ankyrin repeats domains. PIKE-A directly binds to activated Akt but not PI 3-kinase in a guanine nucleotide-dependent way and stimulates the kinase activity of Akt. Overexpression of PIKE-A enhances Akt activity and promotes cancer cell invasion, whereas dominant-negative PIKE-A and PIKE-A knockdown markedly inhibit these processes. Our results demonstrate that PIKE-A is a physiologic regulator of Akt and an oncogenic effector of cell invasion. PMID- 14761977 TI - MDM2 is a negative regulator of p21WAF1/CIP1, independent of p53. AB - The MDM2 oncogene has both p53-dependent and p53-independent activities. We have previously reported that antisense MDM2 inhibitors have significant anti-tumor activity in multiple human cancer models with various p53 statuses (Zhang, Z., Li, M., Wang, H., Agrawal, S., and Zhang, R. (2003) Proc. Natl. Acad. Sci. U. S. A. 100, 11636-11641). We have also provided evidence that MDM2 has a direct role in the regulation of p21, a cyclin-dependent kinase inhibitor. Here we provide evidence supporting functional interaction between MDM2 and p21 in vitro and in vivo. The inhibition of MDM2 with anti-MDM2 antisense oligonucleotide or Short Interference RNA targeting MDM2 significantly elevated p21 protein levels in PC3 cells (p53 null). In contrast, overexpression of MDM2 diminished the p21 level in the same cells by shortening the p21 half-life, an effect reversed by MDM2 antisense inhibition. MDM2 facilitates p21 degradation independent of ubiquitination and the E3 ligase function of MDM2. Instead, MDM2 promotes p21 degradation by facilitating binding of p21 with the proteasomal C8 subunit. The physical interaction between p21 and MDM2 was demonstrated both in vitro and in vivo with the binding region in amino acids 180-298 of the MDM2 protein. In summary, we provide evidence supporting a physical interaction between MDM2 and p21. We also demonstrate that, by reducing p21 protein stability via proteasome mediated degradation, MDM2 functions as a negative regulator of p21, an effect independent of both p53 and ubiquitination. PMID- 14761979 TI - Ald6p is a preferred target for autophagy in yeast, Saccharomyces cerevisiae. AB - Macroautophagy is the process of intracellular bulk protein degradation induced by nutrient starvation and is generally considered to be a nonselective degradation of cytosolic enzymes and organelles. However, it remains a possibility that some proteins may be preferentially degraded by autophagy. In this study, we have performed a systematic analysis on the substrate selectivity of autophagy in yeast, Saccharomyces cerevisiae, using two-dimensional PAGE. We performed a differential screen on wild-type and Deltaatg7/apg7 autophagy deficient cells and found that cytosolic acetaldehyde dehydrogenase (Ald6p) decreased under nitrogen starvation. As assessed by immunoblot, Ald6p was reduced by greater than 82% after 24 h of nitrogen starvation. This reduction was dependent on Atg/Apg proteins and vacuolar proteases but was not dependent on the proteasome or the cytoplasm to vacuole targetting (Cvt) pathway. Using pulse chase and subcellular fractionation, we have also demonstrated that Ald6p was preferentially transported to vacuoles via autophagosomes. Deltaatg7 Deltaald6 double mutant cells were able to maintain higher rates of viability than Deltaatg7 cells under nitrogen starvation, and Ald6p-overexpressing cells were not able to maintain high rates of viability. Furthermore, the Ald6p(C306S) mutant, which lacks enzymatic activity, had viability rates similar to Deltaald6 cells. Ald6p enzymatic activity may be disadvantageous for survival under nitrogen starvation; therefore, yeast cells may preferentially eliminate Ald6p via autophagy. PMID- 14761978 TI - Dual role of Rac in the assembly of NADPH oxidase, tethering to the membrane and activation of p67phox: a study based on mutagenesis of p67phox-Rac1 chimeras. AB - NADPH oxidase activation involves the assembly of membrane-localized cytochrome b559 with the cytosolic components p47phox, p67phox, and the small GTPase Rac. Assembly is mimicked by a cell-free system consisting of membranes and cytosolic components, activated by an anionic amphiphile. We reported that a chimeric construct, consisting of residues 1-212 of p67phox and full-length Rac1, activates the oxidase in vitro in an amphiphile-dependent manner, and when prenylated, in the absence of amphiphile and p47phox. We subjected chimera p67phox-(1-212)-Rac1 to mutational analysis and found that: 1) replacement of a single basic residue at the C terminus of the Rac1 moiety by glutamine is sufficient for loss of activity by the non-prenylated chimera; replacement of all six basic residues by glutamines is required for loss of activity by the prenylated chimera. 2) A V204A mutation in the activation domain of the p67phox moiety leads to a reduction in activity. 3) Mutating residues, known to participate in the interaction between free p67phox and Rac1, in the p67phox (R102E) or Rac1 (A27K, G30S) moieties of the chimera, leads to a marked decrease in activity, indicating a requirement for intrachimeric bonds, in addition to the engineered fusion. 4) Chimeras, inactive because of mutations A27K or G30S in the Rac1 moiety, are reactivated by supplementation with exogenous Rac1-GTP but not with exogenous p67phox. This demonstrates that Rac has a dual role in the assembly of NADPH oxidase. One is to tether p67phox to the membrane; the other is to induce an "activating" conformational change in p67phox. PMID- 14761980 TI - The Pro-451 to Leu polymorphism within the C-terminal tail of P2X7 receptor impairs cell death but not phospholipase D activation in murine thymocytes. AB - The P2X family of ATP receptors (P2XR) are ligandgated channels that have been proposed to regulate cell death of immature thymocytes. However, the nature of the P2XR subtype involved has been controversial until recently. In agreement with previous studies, we found that extracellular ATP (ATPe) induces a caspase dependent apoptosis of BALB/c thymocytes, as observed by DNA fragmentation. Additionally, ATPe induces a predominant caspase-independent thymocytes lysis characterized by plasma membrane disruption. Both responses to ATPe can be induced by a potent P2X7R agonist, benzoylbenzoyl-ATP, whereas P2X7R antagonists, oxidized ATP and pyridoxalphosphate-6-azophenyl-2',4'-disulfonic acid, inhibited the effect of ATPe. These results are further supported by observations where disruption of the P2X7R gene (P2X7R(-/-) mice) completely abolishes thymocytes death induced by ATPe. Interestingly, the natural P451L mutation in the C terminal tail of P2X7R present in C57BL/6 mice, which impairs ATPe-dependent pore formation in T lymphocytes, significantly reduces thymocytes death triggered by ATPe. Furthermore, we found that P2X7R from BW5147 thymoma cells also harbors this point mutation, accounting for their insensitivity to ATPe-induced cell death. Concentrations of ATPe effective in inducing cell death also increase phosphatidylcholine-hydrolyzing phospholipase D (PC-PLD) activity in BALB/c thymocytes through the stimulation of P2X7R. However, in contrast to ATPe-induced cell death, PC-PLD activation is totally Ca(2+)-dependent. Moreover, the stimulation of PC-PLD by ATPe is not affected by the P451L mutation present in C57BL/6 thymocytes and BW5147 cells, suggesting that cell death and PC-PLD activity are regulated through distinct domains of the P2X7R. Finally, the inhibition of ATPe-induced PC-PLD stimulation does not affect thymocytes death. Altogether, these data suggest that P2X7R-induced thymocytes death is independent of the stimulation of PC-PLD activity. PMID- 14761981 TI - Oncogenic activation of c-Myb correlates with a loss of negative regulation by TIF1beta and Ski. AB - The c-myb proto-oncogene product (c-Myb) regulates proliferation of hematopoietic cells by inducing the transcription of a group of target genes. Removal or mutations of the negative regulatory domain (NRD) in the C-terminal half of c-Myb leads to increased transactivating capacity and oncogenic activation. Here we report that TIF1beta directly binds to the NRD and negatively regulates the c-Myb dependent trans-activation. In addition, three corepressors (Ski, N-CoR, and mSin3A) bind to the DNA-binding domain of c-Myb together with TIF1beta and recruit the histone deacetylase complex to c-Myb. Furthermore, the Drosophila TIF1beta homolog, Bonus, negatively regulates Drosophila Myb activity. The Ski corepressor competes with the coactivator CBP for binding to c-Myb, indicating that the selection of coactivators and corepressors is a key event for c-Myb dependent transcription. Mutations or deletion of the NRD of c-Myb and the mutations found in the DNA-binding domain of v-Myb decrease the interaction with these corepressors and weaken the corepressor-induced negative regulation of Myb activity. These observations have conceptual implications for understanding how the nuclear oncogene is activated. PMID- 14761982 TI - Influence of lateral association on forced unfolding of antiparallel spectrin heterodimers. AB - Protein extensibility appears to be based broadly on conformational changes that can in principle be modulated by protein-protein interactions. Spectrin family proteins, with their extensible three-helix folds, enable evaluation of dimerization effects at the single molecule level by atomic force microscopy. Although some spectrin family members function physiologically only as homodimers (e.g. alpha-actinin) or are strictly monomers (e.g. dystrophin), alpha- and beta spectrins are stable as monomeric forms but occur physiologically as alpha,beta heterodimers bound laterally lengthwise. For short constructs of alpha- and beta spectrin, either as monomers or as alpha,beta-dimers, sawtooth patterns in atomic force microscopy-forced extension show that unfolding stochastically extends repeats approximately 4-5-fold greater in length than native conformations. For both dimers and monomers, distributions of unfolding lengths appear bimodal; major unfolding peaks reflect single repeats, and minor unfolding peaks at twice the length reflect tandem repeats. Cooperative unfolding thus propagates through helical linkers between serial repeats (1, 2). With lateral heterodimers, however, the force distribution is broad and shifted to higher forces. The associated chains in a dimer can stay together and unfold simultaneously in addition to unfolding independently. Weak lateral interactions do not inhibit unfolding, but strong lateral interactions facilitate simultaneous unfolding analogous to serial repeat coupling within spectrin family proteins. PMID- 14761983 TI - Sigma factors and global gene regulation in Mycobacterium tuberculosis. PMID- 14761984 TI - Functional genomics of gram-positive microorganisms. PMID- 14761985 TI - H-NS represses Salmonella enterica serovar Typhimurium dsbA expression during exponential growth. AB - Disulfide bond formation catalyzed by disulfide oxidoreductases occurs in the periplasm and plays a major role in the proper folding and integrity of many proteins. In this study, we were interested in elucidating factors that influence the regulation of dsbA, a gene coding for the primary disulfide oxidoreductase found in Salmonella enterica serovar Typhimurium. Strains with mutations created by transposon mutagenesis were screened for strains with altered expression of dsbA. A mutant (NLM2173) was found where maximal expression of a dsbA::lacZ transcriptional fusion occurred in the exponential growth phase in contrast to that observed in the wild type where maximal expression occurs in stationary phase. Sequence analysis of NLM2173 demonstrated that the transposon had inserted upstream of the gene encoding H-NS. Western immunoblot analysis using H-NS and StpA antibodies showed decreased amounts of H-NS protein in NLM2173, and this reduction in H-NS correlated with an increase of StpA protein. Northern blot analysis with a dsbA-specific probe showed an increase in dsbA transcript during exponential phase of growth. Direct binding of H-NS to the dsbA promoter region was verified using purified H-NS in electrophoretic mobility shift assays. Thus, a reduction in H-NS protein is correlated with a derepression of dsbA in NLM2173, suggesting that H-NS normally plays a role in suppressing the expression of dsbA during exponential phase growth. PMID- 14761986 TI - Dynamics of fruiting body morphogenesis. AB - Myxobacteria build their species-specific fruiting bodies by cell movement and then differentiate spores in specific places within that multicellular structure. New steps in the developmental aggregation of Myxococcus xanthus were discovered through a frame-by-frame analysis of a motion picture. The formation and fate of 18 aggregates were captured in the time-lapse movie. Still photographs of 600 other aggregates were also analyzed. M. xanthus has two engines that propel the gliding of its rod-shaped cells: slime-secreting jets at the rear and retractile pili at the front. The earliest aggregates are stationary masses of cells that look like three-dimensional traffic jams. We propose a model in which both engines stall as the cells' forward progress is blocked by other cells in the traffic jam. We also propose that these blockades are eventually circumvented by the cell's capacity to turn, which is facilitated by the push of slime secretion at the rear of each cell and by the flexibility of the myxobacterial cell wall. Turning by many cells would transform a traffic jam into an elliptical mound, in which the cells are streaming in closed orbits. Pairs of adjacent mounds are observed to coalesce into single larger mounds, probably reflecting the fusion of orbits in the adjacent mounds. Although fruiting bodies are relatively large structures that contain 10(5) cells, no long-range interactions between cells were evident. For aggregation, M. xanthus appears to use local interactions between its cells. PMID- 14761987 TI - Inhibition of membrane-bound methane monooxygenase and ammonia monooxygenase by diphenyliodonium: implications for electron transfer. AB - Diphenyliodonium (DPI) is known to irreversibly inactivate flavoproteins. We have found that DPI inhibits both membrane-bound methane monooxygenase (pMMO) from Methylococcus capsulatus and ammonia monooxygenase (AMO) of Nitrosomonas europaea. The effect of DPI on NADH-dependent pMMO activity in vitro is ascribed to inactivation of NDH-2, a flavoprotein which we proposed catalyzes reduction of the quinone pool by NADH. DPI is a potent inhibitor of type 2 NADH:quinone oxidoreductase (NDH-2), with 50% inhibition occurring at approximately 5 micro M. Inhibition of NDH-2 is irreversible and requires NADH. Inhibition of NADH dependent pMMO activity by DPI in vitro is concomitant with inhibition of NDH-2, consistent with our proposal that NDH-2 mediates reduction of pMMO. Unexpectedly, DPI also inhibits pMMO activity driven by exogenous hydroquinols, but with approximately 100 micro M DPI required to achieve 50% inhibition. Similar concentrations of DPI are required to inhibit formate-, formaldehyde-, and hydroquinol-driven pMMO activities in whole cells. The pMMO activity in DPI treated cells greatly exceeds the activity of NDH-2 or pMMO in membranes isolated from those cells, suggesting that electron transfer from formate to pMMO in vivo can occur independent of NADH and NDH-2. AMO activity, which is known to be independent of NADH, is affected by DPI in a manner analogous to pMMO in vivo: approximately 100 micro M is required for 50% inhibition regardless of the nature of the reducing agent. DPI does not affect hydroxylamine oxidoreductase activity and does not require AMO turnover to exert its inhibitory effect. Implications of these data for the electron transfer pathway from the quinone pool to pMMO and AMO are discussed. PMID- 14761988 TI - Integration of environmental signals controls expression of Bordetella heme utilization genes. AB - The Bordetella pertussis heme utilization gene cluster hurIR bhuRSTUV encodes regulatory and transport functions required for assimilation of iron from heme and hemoproteins. Expression of the bhu genes is iron regulated and heme inducible. The putative extracytoplasmic function (ECF) sigma factor, HurI, is required for heme-responsive bhu gene expression. In this study, transcriptional activation of B. pertussis bhu genes in response to heme compounds was shown to be dose dependent and specific for heme; protoporphyrin IX and other heme structural analogs did not activate bhu gene expression. Two promoters controlling expression of the heme utilization genes were mapped by primer extension analysis. The hurI promoter showed similarity to sigma(70)-like promoters, and its transcriptional activity was iron regulated and heme independent. A second promoter identified upstream of bhuR exhibited little similarity to previously characterized ECF sigma factor-dependent promoters. Expression of bhuR was iron regulated, heme responsive, and hurI dependent in B. pertussis, as shown in a previous study with Bordetella bronchiseptica. Further analyses showed that transcription originating at a distal upstream site and reading through the hurR-bhuR intergenic region contributes to bhuR expression under iron starvation conditions in the absence of heme inducer. The pattern of regulation of the readthrough transcript was consistent with transcription from the hurI promoter. The positions and regulation of the two promoters within the hur-bhu gene cluster influence the production of heme transport machinery so that maximal expression of the bhu genes occurs under iron starvation conditions only in the presence of heme iron sources. PMID- 14761989 TI - Proteomic analysis of the sarcosine-insoluble outer membrane fraction of Helicobacter pylori strain 26695. AB - Helicobacter pylori causes gastroduodenal disease, which is mediated in part by its outer membrane proteins (OMPs). To identify OMPs of H. pylori strain 26695, we performed a proteomic analysis. A sarcosine-insoluble outer membrane fraction was resolved by two-dimensional electrophoresis with immobilized pH gradient strips. Most of the protein spots, with molecular masses of 10 to 100 kDa, were visible on the gel in the alkaline pI regions (6.0 to 10.0). The proteome of the OMPs was analyzed by matrix-assisted laser desorption ionization-time-of-flight mass spectrometry. Of the 80 protein spots processed, 62 spots were identified; they represented 35 genes, including 16 kinds of OMP. Moreover, we identified 9 immunoreactive proteins by immunoblot analysis. This study contributes to the characterization of the H. pylori strain 26695 proteome and may help to further elucidate the biological function of H. pylori OMPs and the pathogenesis of H. pylori infection. PMID- 14761990 TI - The sufR gene (sll0088 in Synechocystis sp. strain PCC 6803) functions as a repressor of the sufBCDS operon in iron-sulfur cluster biogenesis in cyanobacteria. AB - The suf operon is composed of four genes (sufB, sufC, sufD, and sufS) and is highly conserved in the genomes of cyanobacteria. Open reading frame sll0088 in Synechocystis sp. strain PCC 6803 is located near the 5' end of the suf operon but is transcribed in the direction opposite that of the suf operon. We previously reported the isolation of two independent suppressor strains of C14S(PsaC) that mapped to sll0088 and restored photoautotrophic growth. The protein encoded by sll0088 has two significant features: (i) a DNA-binding domain near the N terminus and (ii) four highly conserved cysteine residues near the C terminus. The protein has high sequence similarity to transcription regulatory proteins with a conserved DNA-binding domain and can be classified in the DeoR family of helix-loop-helix proteins. The protein falls into a further subclass that contains a C-X(12)-C-X(13)-C-X(14)-C motif near the C terminus, which may represent a metal-binding site. The expressed Sll0088 protein harbored an iron sulfur cluster as shown by optical and electron paramagnetic resonance spectroscopy. Compared to the wild type, expression levels of the sufBCDS genes were elevated when cells were grown under conditions of oxidative and iron stress and were even higher in a null mutant of Synechococcus sp. strain PCC 7002 in which the sll0088 homolog was insertionally inactivated. In agreement with the proposed role of the sufBCDS genes in iron metabolism, the growth rate of the null mutant was significantly higher than that of the wild type under iron limiting conditions. We propose that the protein encoded by sll0088 is a transcriptional repressor of the suf operon, and we name the gene sufR. PMID- 14761991 TI - Architecture of the Bacteroides cellulosolvens cellulosome: description of a cell surface-anchoring scaffoldin and a family 48 cellulase. AB - A large gene downstream of the primary Bacteroides cellulosolvens cellulosomal scaffoldin (cipBc, now renamed scaA) was sequenced. The gene, termed scaB, contained an N-terminal leader peptide followed by 10 type I cohesins, an "X" domain of unknown structure and function, and a C-terminal S-layer homology (SLH) surface-anchoring module. In addition, a previously identified gene in a different part of the genome, encoding for a dockerin-borne family 48 cellulosomal glycoside hydrolase (Cel48), was sequenced completely, and a putative cellulosome-related family 9 glycosyl hydrolase was detected. Recombinant fusion proteins, comprising dockerins derived from either the ScaA scaffoldin or Cel48, were overexpressed. Their interaction with ScaA and ScaB cohesins was examined by immunoassay. The results indicated that the ScaB type I cohesin of the new anchoring protein binds selectively to the ScaA dockerin, whereas the Cel48 dockerin binds specifically to the type II ScaA cohesin 5. Thus, by virtue of the 11 type II ScaA cohesins and the 10 type I ScaB cohesins, the relatively simple two-component cellulosome-integrating complex would potentially incorporate 110 enzyme molecules onto the cell surface via the ScaB SLH module. Compared to previously described cellulosome systems, the apparent roles of the B. cellulosolvens cohesins are reversed, in that the type II cohesins are located on the enzyme-binding primary scaffoldin, whereas the type I cohesins are located on the anchoring scaffoldin. The results underscore the extensive diversity in the supramolecular architecture of cellulosome systems in nature. PMID- 14761992 TI - Genetic and structural characterization of the core region of the lipopolysaccharide from Serratia marcescens N28b (serovar O4). AB - The gene cluster (waa) involved in Serratia marcescens N28b core lipopolysaccharide (LPS) biosynthesis was identified, cloned, and sequenced. Complementation analysis of known waa mutants from Escherichia coli K-12, Salmonella enterica, and Klebsiella pneumoniae led to the identification of five genes coding for products involved in the biosynthesis of a shared inner core structure: [L,D-HeppIIIalpha(1-->7)-L,D-HeppIIalpha(1-->3)-L,D-HeppIalpha(1-->5) KdopI(4<--2)alphaKdopII] (L,D-Hepp, L-glycero-D-manno-heptopyranose; Kdo, 3-deoxy D-manno-oct-2-ulosonic acid). Complementation and/or chemical analysis of several nonpolar mutants within the S. marcescens waa gene cluster suggested that in addition, three waa genes were shared by S. marcescens and K. pneumoniae, indicating that the core region of the LPS of S. marcescens and K. pneumoniae possesses additional common features. Chemical and structural analysis of the major oligosaccharide from the core region of LPS of an O-antigen-deficient mutant of S. marcescens N28b as well as complementation analysis led to the following proposed structure: beta-Glc-(1-->6)-alpha-Glc-(1-->4))-alpha-D-GlcN-(1 ->4)-alpha-D-GalA-[(2<--1)-alpha-D,D-Hep-(2<--1)-alpha-Hep]-(1-->3)-alpha-L,D Hep[(7<--1)-alpha-L,D-Hep]-(1-->3)-alpha-L,D-Hep-[(4<--1)-beta-D-Glc]-(1-->5) Kdo. The D configuration of the beta-Glc, alpha-GclN, and alpha-GalA residues was deduced from genetic data and thus is tentative. Furthermore, other oligosaccharides were identified by ion cyclotron resonance-Fourier-transformed electrospray ionization mass spectrometry, which presumably contained in addition one residue of D-glycero-D-talo-oct-2-ulosonic acid (Ko) or of a hexuronic acid. Several ions were identified that differed from others by a mass of +80 Da, suggesting a nonstoichiometric substitution by a monophosphate residue. However, none of these molecular species could be isolated in substantial amounts and structurally analyzed. On the basis of the structure shown above and the analysis of nonpolar mutants, functions are suggested for the genes involved in core biosynthesis. PMID- 14761993 TI - Novel roles of the master transcription factors Spo0A and sigmaB for survival and sporulation of Bacillus subtilis at low growth temperature. AB - Spore development and stress resistance in Bacillus subtilis are governed by the master transcription factors Spo0A and sigma(B), respectively. Here we show that the coding genes for both regulatory proteins are dramatically induced, during logarithmic growth, after a temperature downshift from 37 to 20 degrees C. The loss of sigma(B) reduces the stationary-phase viability of cold-adapted cells 10- to 50-fold. Furthermore, we show that sigma(B) activity is required at a late stage of development for efficient sporulation at a low temperature. On the other hand, Spo0A loss dramatically reduces the stationary-phase viability of cold adapted cells 10,000-fold. We show that the requirement of Spo0A for cellular survival during the cold is independent of the activity of the key transition state regulator AbrB and of the simple loss of sporulation ability. Furthermore, Spo0A, and not proficiency in sporulation, is required for the development of complete stress resistance of cold-adapted cells to heat shock (54 degrees C, 1 h), since a loss of Spo0A, but not a loss of the essential sporulation transcription factor sigma(F), reduced the cellular survival in response to heat by more than 1,000-fold. The overall results argue for new and important roles for Spo0A in the development of full stress resistance by nonsporulating cells and for sigma(B) in sporulation proficiency at a low temperature. PMID- 14761994 TI - Myxococcus xanthus chemotaxis homologs DifD and DifG negatively regulate fibril polysaccharide production. AB - The extracellular matrix fibrils of Myxococcus xanthus are essential for the social lifestyle of this unusual bacterium. These fibrils form networks linking or encasing cells and are tightly correlated with cellular cohesion, development, and social (S) gliding motility. Previous studies identified a set of bacterial chemotaxis homologs encoded by the dif locus. It was determined that difA, difC, and difE, encoding respective homologs of a methyl-accepting chemotaxis protein, CheW, and CheA, are required for fibril production and therefore S motility and development. Here we report the studies of three additional genes residing at the dif locus, difB, difD, and difG. difD and difG encode homologs of chemotaxis proteins CheY and CheC, respectively. difB encodes a positively charged protein with limited homology at its N terminus to conserved bacterial proteins with unknown functions. Unlike the previously characterized dif genes, none of these three newly studied dif genes are essential for fibril production, S motility, or development. The difB mutant showed no obvious defects in any of the processes examined. In contrast, the difD and the difG mutants were observed to overproduce fibril polysaccharides in comparison with production by the wild type. The observation that DifD and DifG negatively regulate fibril polysaccharide production strengthens our hypothesis that the M. xanthus dif genes define a chemotaxis-like signal transduction pathway which regulates fibril biogenesis. To our knowledge, this is the first report of functional studies of a CheC homolog in proteobacteria. In addition, during this study, we slightly modified previously developed assays to easily quantify fibril polysaccharide production in M. xanthus. PMID- 14761995 TI - Transcriptional and posttranscriptional control of cable pilus gene expression in Burkholderia cenocepacia. AB - Burkholderia cenocepacia is an important member of the Burkholderia cepacia complex, a group of closely related bacteria that inhabits a wide variety of environmental niches in nature and that also colonizes the lungs of compromised humans. Certain strains of B. cenocepacia express peritrichous adherence organelles known as cable pili, thought to be important in the colonization of the lower respiratory tract. The genetic locus required for cable pilus biogenesis is comprised of at least five genes, designated cblB, cblA, cblC, cblD, and cblS. In this study a transcriptional analysis of cbl gene expression was undertaken. The principal promoter, located upstream of the cbl locus, was identified and characterized. By using lacZ transcriptional fusions, the effects of multiple environmental cues on cbl gene expression were examined. High osmolarity, temperature of 37 degrees C, acidic pH, and low iron bioavailability were found to induce cbl gene expression. Northern hybridization analysis of the cbl locus identified a single, stable transcript corresponding to cblA, encoding the major pilin subunit. Transcriptional fusion studies combined with reverse transcription-PCR analysis indicated that the stable cblA transcript is the product of an mRNA processing event. This event may ensure high levels of expression of the major pilin, relative to other components of the assembly pathway. Our findings lend further insight into the control of cable pilus biogenesis in B. cenocepacia and provide evidence for regulation of cbl gene expression on both the transcriptional and posttranscriptional levels. PMID- 14761996 TI - OppA, the substrate-binding subunit of the oligopeptide permease, is the major Ecto-ATPase of Mycoplasma hominis. AB - Most ATPases, involved in energy-driven processes, act in the cytoplasm. However, external membrane-bound ATPases have also been described in parasites and eukaryotic cells. In Mycoplasma hominis, a bacterium lacking a cell wall, the surface-exposed substrate-binding protein OppA of an oligopeptide permease (Opp) contains an ATP binding P-loop structure in the C-terminal region. With ATP affinity chromatography and tryptic digestion in the presence or absence of ATP, the functionality of the Mg(2+)-dependent ATP binding site is demonstrated. In addition to ATP, ADP also could bind to OppA. The presence of an ATPase activity on the surface of M. hominis is indicated by the inactivation of ATP hydrolyzing activity of intact mycoplasma cells by the impermeable ATPase inhibitor 4',4' diisothiocyanostilbene-2',2'-disulfonic acid and influenced by the ATP analog 5' fluorosulfonyl-benzoyladenosine. Comparing equimolar amounts of OppA in intact mycoplasma cells and in the purified form indicated that more than 80% of the surface-localized ATPase activity is derived from OppA, implying that OppA is the main ATPase on the surface of mycoplasma cells. Together, these data present the first evidence that the cytoadhesive substrate binding protein OppA of the oligopeptide permease also functions as an ecto-ATPase in Mycoplasma hominis. PMID- 14761997 TI - Molecular characterization of a high-affinity xylobiose transporter of Streptomyces thermoviolaceus OPC-520 and its transcriptional regulation. AB - Streptomyces thermoviolaceus OPC-520 secretes two types of xylanases (StxI and StxII), an acetyl xylan esterase (StxIII), and an alpha-L-arabinofuranosidase (StxIV) in the presence of xylan. Xylan degradation products (mainly xylobiose) produced by the action of these enzymes entered the cell and were then degraded to xylose by an intracellular beta-xylosidase (BxlA). A gene cluster involved in xylanolytic system of the strain was cloned and sequenced upstream of and including a BxlA-encoding gene (bxlA). The gene cluster consisted of four different open reading frames organized in the order bxlE, bxlF, bxlG, and bxlA. Reverse transcriptase PCR analysis revealed that the gene cluster is transcribed as polycistronic mRNA. The deduced gene products, comprising BxlE (a sugar binding lipoprotein), BxlF (an integral membrane protein), and BxlG (an integral membrane protein), showed similarity to components of the bacterial ATP-binding cassette (ABC) transport system; however, the gene for the ATP binding protein was not linked to the bxl operon. The soluble recombinant BxlE protein was analyzed for its binding activity for xylooligosaccharides. The protein showed high-level affinity for xylobiose (K(d) = 8.75 x 10(-9) M) and for xylotriose (K(d) = 8.42 x 10(-8) M). Antibodies raised against the recombinant BxlE recognized the detergent-soluble BxlE isolated from S. thermoviolaceus membranes. The deduced BxlF and BxlG proteins are predicted to be integral membrane proteins. These proteins contained the conserved EAA loop (between the fourth and the fifth membrane-spanning segments) which is characteristic of membrane proteins from binding-protein-dependent ABC transporters. In addition, the bxlR gene located upstream of the bxl operon was cloned and expressed in Escherichia coli. The bxlR gene encoded a 343-residue polypeptide that is highly homologous to members of the GalR/LacI family of bacterial transcriptional regulators. The purified BxlR protein specifically bound to a 4-bp inverted sequence overlapping the -10 region of the bxl operon. The binding of BxlR to the site was inhibited specifically by low concentrations of xylobiose. This site was also present in the region located between stxI and stxIV and in the upstream region of stxII. BxlR specifically bound to the regions containing the inverted sequence. These results suggest that BxlR might act as a repressor of the genes involved not only in the uptake system of xylan degradation products but also in xylan degradation of S. thermoviolaceus OPC-520. PMID- 14761998 TI - The lgtABCDE gene cluster, involved in lipooligosaccharide biosynthesis in Neisseria gonorrhoeae, contains multiple promoter sequences. AB - Biosynthesis of the variable core domain of lipooligosaccharide (LOS) in Neisseria gonorrhoeae is mediated by glycosyl transferases encoded by lgtABCDE. Changes within homopolymeric runs within lgtA, lgtC, and lgtD affect the expression state of these genes, with the nature of the LOS expressed determined by the functionality of these genes. However, the mechanism for modulating the amount of multiple LOS chemotypes expressed in a single cell is not understood. Using mutants containing polar disruptions within the lgtABCDE locus, we determined that the expression of this locus is mediated by multiple promoters and that disruption of transcription from these promoters alters the relative levels of simultaneously expressed LOS chemotypes. Expression of the lgtABCDE locus was quantified by using xylE transcriptional fusions, and the data indicate that this locus is transcribed in trace amounts and that subtle changes in transcription result in phenotypic changes. By using rapid amplification of 5' cDNA ends, transcriptional start sites and promoter sequences were identified within lgtABCDE. Most of these promoters possessed 50 to 67% homology with the consensus gearbox promoter sequence of Escherichia coli. PMID- 14761999 TI - The ytkD (mutTA) gene of Bacillus subtilis encodes a functional antimutator 8-Oxo (dGTP/GTP)ase and is under dual control of sigma A and sigma F RNA polymerases. AB - The regulation of expression of ytkD, a gene that encodes the first functional antimutator 8-oxo-dGTPase activity of B. subtilis, was studied here. A ytkD-lacZ fusion integrated into the ytkD locus of wild-type B. subtilis 168 revealed that this gene is expressed during both vegetative growth and early stages of sporulation. In agreement with this result, ytkD mRNAs were detected by both Northern blotting and reverse transcription-PCR during both developmental stages. These results suggested that ytkD is transcribed by the sequential action of RNA polymerases containing the sigma factors sigma(A) and sigma(F), respectively. In agreement with this suggestion, the spore-associated expression was almost completely abolished in a sigF genetic background but not in a B. subtilis strain lacking a functional sigG gene. Primer extension analysis mapped transcriptional start sites on mRNA samples isolated from vegetative and early sporulating cells of B. subtilis. Inspection of the sequences lying upstream of the transcription start sites revealed the existence of typical sigma(A)- and sigma(F)-type promoters. These results support the conclusion that ytkD expression is subjected to dual regulation and suggest that the antimutator activity of YtkD is required not only during vegetative growth but also during the early sporulation stages and/or germination of B. subtilis. While ytkD expression obeyed a dual pattern of temporal expression, specific stress induction of the transcription of this gene does not appear to occur, since neither oxidative damage (following either treatment with paraquat or hydrogen peroxide) nor mitomycin C treatment or sigma(B) general stress inducers (sodium chloride, ethanol, or heat) affected the levels of the gene product produced. PMID- 14762000 TI - Evolution of Staphylococcus aureus by large chromosomal replacements. AB - Conjugative transfer and replacement of hundreds of kilobases of a bacterial chromosome can occur in vitro, but replacements in nature are either an order of magnitude smaller or involve the movement of mobile genetic elements. We discovered that two lineages of Staphylococcus aureus, including a pandemic methicillin-resistant lineage, were founded by single chromosomal replacements of at least approximately 244 and approximately 557 kb representing approximately 10 and approximately 20% of the chromosome, respectively, without the obvious involvement of mobile genetic elements. The replacements are unprecedented in natural populations of bacteria because of their large size and unique structure and may have a dramatic impact on bacterial evolution. PMID- 14762001 TI - Real-time monitoring of intracellular Staphylococcus aureus replication. AB - A high-throughput system to rapidly assess the intracellular replication of Staphylococcus aureus has been developed utilizing S. aureus transformed with a dual gfp-luxABCDE reporter operon under the control of a growth-dependent promoter. Replication of tagged bacteria internalized into bovine mammary epithelial cells (MAC-T) could be measured by monitoring fluorescence and bioluminescence from the reporter operon following removal of extracellular bacteria from the plates. Bacterial replication inside cells was confirmed by a novel ex vivo time-lapse confocal microscopic method. This assay of bacterial replication was used to evaluate the efficacy of antibiotics which are commonly used to treat staphylococcal infections. Not all antibiotics tested were able to prevent intracellular replication of S. aureus and some were ineffective at preventing replication of intracellular bacteria at concentrations above the MIC determined for bacteria in broth culture. Comparison of the fluorescence and bioluminescence signals from the bacteria enabled effects on protein synthesis and metabolism to be discriminated and gave information on the entry of compounds into the eukaryotic cell, even if bacterial replication was not prevented. Elevated resistance of S. aureus to antibiotics inside host cells increases the likelihood of selecting S. aureus strains which are resistant to commonly used antimicrobial agents within the intracellular niche. The approach presented directly assesses intracellular efficacy of antibiotics and provides an evidence based approach to antibiotic selection for prescribing physicians and medical microbiologists. PMID- 14762002 TI - Alpha-helix E of Spo0A is required for sigmaA- but not for sigmaH-dependent promoter activation in Bacillus subtilis. AB - At the onset of endospore formation in Bacillus subtilis, the DNA binding protein Spo0A activates transcription from two types of promoters. The first type includes the spoIIG and spoIIE promoters, which are used by sigma(A)-RNA polymerase, whereas the second type includes the spoIIA promoter, which is used by RNA polymerase containing the secondary sigma factor sigma(H). Previous genetic analyses have identified specific amino acids in alpha-helix E of Spo0A that are important for activation of Spo0A-dependent, sigma(A)-dependent promoters. However, these amino acids are not required for activation of the sigma(H)-dependent spoIIA promoter. We now report the effects of additional single-amino-acid substitutions and the effects of deletions in alpha-helix E. The effects of alanine substitutions revealed one new position (239) in Spo0A that appears to be specifically required for activation of the sigma(A)-dependent promoters. Based on the effects of a deletion mutation, we suggest that alpha helix E in Spo0A is not directly involved in interaction with sigma(H)-RNA polymerase. PMID- 14762003 TI - Structural and functional characterization of gene clusters directing nonribosomal synthesis of bioactive cyclic lipopeptides in Bacillus amyloliquefaciens strain FZB42. AB - The environmental strain Bacillus amyloliquefaciens FZB42 promotes plant growth and suppresses plant pathogenic organisms present in the rhizosphere. We sampled sequenced the genome of FZB42 and identified 2,947 genes with >50% identity on the amino acid level to the corresponding genes of Bacillus subtilis 168. Six large gene clusters encoding nonribosomal peptide synthetases (NRPS) and polyketide synthases (PKS) occupied 7.5% of the whole genome. Two of the PKS and one of the NRPS encoding gene clusters were unique insertions in the FZB42 genome and are not present in B. subtilis 168. Matrix-assisted laser desorption ionization-time of flight mass spectrometry analysis revealed expression of the antibiotic lipopeptide products surfactin, fengycin, and bacillomycin D. The fengycin (fen) and the surfactin (srf) operons were organized and located as in B. subtilis 168. A large 37.2-kb antibiotic DNA island containing the bmy gene cluster was attributed to the biosynthesis of bacillomycin D. The bmy island was found inserted close to the fen operon. The responsibility of the bmy, fen, and srf gene clusters for the production of the corresponding secondary metabolites was demonstrated by cassette mutagenesis, which led to the loss of the ability to produce these peptides. Although these single mutants still largely retained their ability to control fungal spread, a double mutant lacking both bacillomycin D and fengycin was heavily impaired in its ability to inhibit growth of phytopathogenic fungi, suggesting that both lipopeptides act in a synergistic manner. PMID- 14762004 TI - Differential expression of two paralogous genes of Bacillus subtilis encoding single-stranded DNA binding protein. AB - The Bacillus subtilis genome comprises two paralogous single-stranded DNA binding protein (SSB) genes, ssb and ywpH, which show distinct expression patterns. The main ssb gene is strongly expressed during exponential growth and is coregulated with genes encoding the ribosomal proteins S6 and S18. The gene organization rpsF ssb-rpsR as observed in B. subtilis is found in many gram-positive as well as some gram-negative bacteria, but not in Escherichia coli. The ssb gene is essential for cell viability, and like other SSBs its expression is elevated during SOS response. In contrast, the paralogous ywpH gene is transcribed from its own promoter at the onset of stationary phase in minimal medium only. Its expression is ComK dependent and its gene product is required for optimal natural transformation. PMID- 14762005 TI - ISSa4-based differentiation of Streptococcus agalactiae strains and identification of multiple target sites for ISSa4 insertions. AB - A collection of 113 epidemiologically unrelated Streptococcus agalactiae strains were studied (group B streptococcus; GBS): they belonged to different serotypes and were isolated from pregnant women in China and Russia. The insertion sequence ISSa4 was found in 21 of 113 strains (18,6%). All of the strains with ISSa4 belonged to serotypes II and II/c and were characterized by the presence of IS1381 and IS861 as well as the absence of IS1548 and GBSi1. All of the strains with ISSa4 possessed both bca and bac virulence genes coding for alpha and beta antigens, respectively. Among 21 ISSa4-positive strains, 13 different HindIII patterns (D1 to D13) hybridizing with an ISSa4 probe were found. One of them (D13) contained a single HindIII hybridization fragment 6.5 kb in size that was found to be specific for all ISSa4-positive GBS strains. Multiple target sites for insertions of ISSa4 were identified and included a putative pathogenicity island, "housekeeping" genes, and intergenic regions, as well as the genes for hypothetical proteins. No significant similarity was observed in the sequences of the target genes for ISSa4 insertions, in the relative location of the target genes on the chromosome, or the biological functions of the encoded proteins. The possible significance of ISSa4-based differentiation of the strains and the presence of possible "hot spots" for insertions of ISSa4 in GBS genome are discussed. PMID- 14762006 TI - Interactions among CotB, CotG, and CotH during assembly of the Bacillus subtilis spore coat. AB - Spores formed by wild-type Bacillus subtilis are encased in a multilayered protein structure (called the coat) formed by the ordered assembly of over 30 polypeptides. One polypeptide (CotB) is a surface-exposed coat component that has been used as a vehicle for the display of heterologous antigens at the spore surface. The cotB gene was initially identified by reverse genetics as encoding an abundant coat component. cotB is predicted to code for a 43-kDa polypeptide, but the form that prevails in the spore coat has a molecular mass of about 66 kDa (herein designated CotB-66). Here we show that in good agreement with its predicted size, expression of cotB in Escherichia coli results in the accumulation of a 46-kDa protein (CotB-46). Expression of cotB in sporulating cells of B. subtilis also results in a 46-kDa polypeptide which appears to be rapidly converted into CotB-66. These results suggest that soon after synthesis, CotB undergoes a posttranslational modification. Assembly of CotB-66 has been shown to depend on expression of both the cotH and cotG loci. We found that CotB 46 is the predominant form found in extracts prepared from sporulating cells or in spore coat preparations of cotH or cotG mutants. Therefore, both cotH and cotG are required for the efficient conversion of CotB-46 into CotB-66 but are dispensable for the association of CotB-46 with the spore coat. We also show that CotG does not accumulate in sporulating cells of a cotH mutant, suggesting that CotH (or a CotH-controlled factor) stabilizes the otherwise unstable CotG. Thus, the need for CotH for formation of CotB-66 results in part from its role in the stabilization of CotG. We also found that CotB-46 is present in complexes with CotG at the time when formation of CotB-66 is detected. Moreover, using a yeast two-hybrid system, we found evidence that CotB directly interacts with CotG and that both CotB and CotG self-interact. We suggest that an interaction between CotG and CotB is required for the formation of CotB-66, which may represent a multimeric form of CotB. PMID- 14762007 TI - Mechanism of transcription activation at the comG promoter by the competence transcription factor ComK of Bacillus subtilis. AB - The development of genetic competence in Bacillus subtilis is regulated by a complex signal transduction cascade, which results in the synthesis of the competence transcription factor, encoded by comK. ComK is required for the transcription of the late competence genes that encode the DNA binding and uptake machinery and of genes required for homologous recombination. In vivo and in vitro experiments have shown that ComK is responsible for transcription activation at the comG promoter. In this study, we investigated the mechanism of this transcription activation. The intrinsic binding characteristics of RNA polymerase with and without ComK at the comG promoter were determined, demonstrating that ComK stabilizes the binding of RNA polymerase to the comG promoter. This stabilization probably occurs through interactions with the upstream DNA, since a deletion of the upstream DNA resulted in an almost complete abolishment of stabilization of RNA polymerase binding. Furthermore, a strong requirement for the presence of an extra AT box in addition to the common ComK binding site was shown. In vitro transcription with B. subtilis RNA polymerase reconstituted with wild-type alpha-subunits and with C-terminal deletion mutants of the alpha-subunits was performed, demonstrating that these deletions do not abolish transcription activation by ComK. This indicates that ComK is not a type I activator. We also show that ComK is not required for open complex formation. A possible mechanism for transcription activation is proposed, implying that the major stimulatory effect of ComK is on binding of RNA polymerase. PMID- 14762008 TI - Assembly of multiple CotC forms into the Bacillus subtilis spore coat. AB - We report evidence that the CotC polypeptide, a previously identified component of the Bacillus subtilis spore coat, is assembled into at least four distinct forms. Two of these, having molecular masses of 12 and 21 kDa, appeared 8 h after the onset of sporulation and were probably assembled on the forming spore immediately after their synthesis, since no accumulation of either of them was detected in the mother cell compartment, where their synthesis occurs. The other two components, 12.5 and 30 kDa, were generated 2 h later and were probably the products of posttranslational modifications of the two early forms occurring directly on the coat surface during spore maturation. None of the CotC forms was found either on the spore coat or in the mother cell compartment of a cotH mutant. This indicates that CotH serves a dual role of stabilizing the early forms of CotC and promoting the assembly of both early and late forms on the spore surface. PMID- 14762009 TI - The Bacillus subtilis extracytoplasmic-function sigmaX factor regulates modification of the cell envelope and resistance to cationic antimicrobial peptides. AB - Bacillus subtilis contains seven extracytoplasmic-function sigma factors that activate partially overlapping regulons. We here identify four additional members of the sigma(X) regulon, pbpX (penicillin-binding protein), ywnJ, the dlt operon (D-alanylation of teichoic acids), and the pss ybfM psd operon (phosphatidylethanolamine biosynthesis). Modification of teichoic acids by esterification with D-alanine and incorporation of phosphatidylethanolamine into the cell membrane have a common consequence: in both cases positively charged amino groups are introduced into the cell envelope. The resulting reduction in the net negative charge of the cell envelope has been previously implicated as a resistance mechanism specific for cationic antimicrobial peptides. Consistent with this notion, we find that both sigX and dltA mutants are more sensitive to nisin than wild-type cells. We conclude that activation of the sigma(X) regulon serves to alter cell surface properties to provide protection against antimicrobial peptides. PMID- 14762010 TI - ArgR and AhrC are both required for regulation of arginine metabolism in Lactococcus lactis. AB - The DNA binding proteins ArgR and AhrC are essential for regulation of arginine metabolism in Escherichia coli and Bacillus subtilis, respectively. A unique property of these regulators is that they form hexameric protein complexes, mediating repression of arginine biosynthetic pathways as well as activation of arginine catabolic pathways. The gltS-argE operon of Lactococcus lactis encodes a putative glutamate or arginine transport protein and acetylornithine deacetylase, which catalyzes an important step in the arginine biosynthesis pathway. By random integration knockout screening we found that derepression mutants had ISS1 integrations in, among others, argR and ahrC. Single as well as double regulator deletion mutants were constructed from Lactococcus lactis subsp. cremoris MG1363. The three arginine biosynthetic operons argCJDBF, argGH, and gltS-argE were shown to be repressed by the products of argR and ahrC. Furthermore, the arginine catabolic arcABD1C1C2TD2 operon was activated by the product of ahrC but not by that of argR. Expression from the promoter of the argCJDBF operon reached similar levels in the single mutants and in the double mutant, suggesting that the regulators are interdependent and not able to complement each other. At the same time they also appear to have different functions, as only AhrC is involved in activation of arginine catabolism. This is the first study where two homologous arginine regulators are shown to be involved in arginine regulation in a prokaryote, representing an unusual mechanism of regulation. PMID- 14762011 TI - Surface-associated flagellum formation and swarming differentiation in Bacillus subtilis are controlled by the ifm locus. AB - Knowledge of the highly regulated processes governing the production of flagella in Bacillus subtilis is the result of several observations obtained from growing this microorganism in liquid cultures. No information is available regarding the regulation of flagellar formation in B. subtilis in response to contact with a solid surface. One of the best-characterized responses of flagellated eubacteria to surfaces is swarming motility, a coordinate cell differentiation process that allows collective movement of bacteria over solid substrates. This study describes the swarming ability of a B. subtilis hypermotile mutant harboring a mutation in the ifm locus that has long been known to affect the degree of flagellation and motility in liquid media. On solid media, the mutant produces elongated and hyperflagellated cells displaying a 10-fold increase in extracellular flagellin. In contrast to the mutant, the parental strain, as well as other laboratory strains carrying a wild-type ifm locus, fails to activate a swarm response. Furthermore, it stops to produce flagella when transferred from liquid to solid medium. Evidence is provided that the absence of flagella is due to the lack of flagellin gene expression. However, restoration of flagellin synthesis in cells overexpressing sigma(D) or carrying a deletion of flgM does not recover the ability to assemble flagella. Thus, the ifm gene plays a determinantal role in the ability of B. subtilis to contact with solid surfaces. PMID- 14762012 TI - clpB, a novel member of the Listeria monocytogenes CtsR regulon, is involved in virulence but not in general stress tolerance. AB - Clp-HSP100 ATPases are a widespread family of ubiquitous proteins that occur in both prokaryotes and eukaryotes and play important roles in the folding of newly synthesized proteins and refolding of aggregated proteins. They have also been shown to participate in the virulence of several pathogens, including Listeria monocytogenes. Here, we describe a member of the Clp-HSP100 family of L. monocytogenes that harbors all the characteristics of the ClpB subclass, which is absent in the closely related gram-positive model organism, Bacillus subtilis. Transcriptional analysis of clpB revealed a heat shock-inducible sigma(A)-type promoter. Potential binding sites for the CtsR regulator of stress response were identified in the promoter region. In vivo and in vitro approaches were used to show that expression of clpB is repressed by CtsR, a finding indicating that clpB is a novel member of the L. monocytogenes CtsR regulon. We showed that ClpB is involved in the pathogenicity of L. monocytogenes since the DeltaclpB mutant is significantly affected by virulence in a murine model of infection; we also demonstrate that this effect is apparently not due to a defect in general stress resistance. Indeed, ClpB is not involved in tolerance to heat, salt, detergent, puromycin, or cold stress, even though its synthesis is inducible by heat shock. However, ClpB was shown to play a role in induced thermotolerance, allowing increased resistance of L. monocytogenes to lethal temperatures. This work gives the first example of a clpB gene directly controlled by CtsR and describes the first role for a ClpB protein in induced thermotolerance and virulence in a gram positive organism. PMID- 14762013 TI - Identification of genes controlled by the essential YycG/YycF two-component system of Staphylococcus aureus. AB - The YycG/YycF essential two-component system (TCS), originally identified in Bacillus subtilis, is very highly conserved and appears to be specific to low-G+C gram-positive bacteria, including several pathogens such as Staphylococcus aureus. By studying growth of S. aureus cells where the yyc operon is controlled by an isopropyl-beta-D-thiogalactopyranoside (IPTG)-inducible promoter, we have shown that this system is essential in S. aureus during growth at 37 degrees C and that starvation for the YycG/YycF regulatory system leads to cell death. During a previous study of the YycG/YycF TCS of B. subtilis, we defined a potential YycF consensus recognition sequence, consisting of two hexanucleotide direct repeats, separated by five nucleotides [5'-TGT(A/T)A(A/T/C)-N(5) TGT(A/T)A(A/T/C)-3']. A detailed DNA motif analysis of the S. aureus genome indicates that there are potentially 12 genes preceded by this sequence, 5 of which are involved in virulence. An in vitro approach was undertaken to determine which of these genes are controlled by YycF. The YycG and YycF proteins of S. aureus were overproduced in Escherichia coli and purified. Autophosphorylation of the YycG kinase and phosphotransfer to YycF were shown in vitro. Gel mobility shift and DNase I footprinting assays were used to show direct binding in vitro of purified YycF to the promoter region of the ssaA gene, encoding a major antigen and previously suggested to be controlled by YycF. YycF was also shown to bind specifically to the promoter regions of two genes, encoding the IsaA antigen and the LytM peptidoglycan hydrolase, in agreement with the proposed role of this system in controlling virulence and cell wall metabolism. PMID- 14762014 TI - Transcriptional regulation of the phoPR operon in Bacillus subtilis. AB - When Bacillus subtilis is subjected to phosphate starvation, the Pho regulon is activated by the PhoP-PhoR two-component signal transduction system to elicit specific responses to this nutrient limitation. The response regulator, PhoP, and its cognate histidine sensor kinase, PhoR, are encoded by the phoPR operon that is transcribed as a 2.7-kb bicistronic mRNA. The phoPR operon is transcribed from two sigma(A)-dependent promoters, P(1) and P(2). Under conditions where the Pho regulon was not induced (i.e., phosphate-replete conditions or phoR-null mutant), a low level of phoPR transcription was detected only from promoter P(1). During phosphate starvation-induced transition from exponential to stationary phase, the expression of the phoPR operon was up-regulated in a phosphorylated PhoP (PhoP approximately P)-dependent manner; in addition to P(1), the P(2) promoter becomes active. In vitro gel shift assays and DNase I footprinting experiments showed that both PhoP and PhoP approximately P could bind to the control region of the phoPR operon. The data indicate that while low-level constitutive expression of phoPR is required under phosphate-replete conditions for signal perception and transduction, autoinduction is required to provide sufficient PhoP approximately P to induce other members of the Pho regulon. The extent to which promoters P(1) and P(2) are activated appears to be influenced by the presence of other sigma factors, possibly the result of sigma factor competition. For example, phoPR is hyperinduced in a sigB mutant and, later in stationary phase, in sigH, sigF, and sigE mutants. The data point to a complex regulatory network in which other stress responses and post-exponential-phase processes influence the expression of phoPR and, thereby, the magnitude of the Pho regulon response. PMID- 14762015 TI - Physical and enzymological interaction of Bacillus subtilis proteins required for de novo pyridoxal 5'-phosphate biosynthesis. AB - Bacillus subtilis synthesizes pyridoxal 5'-phosphate, the active form of vitamin B(6), by a poorly characterized pathway involving the yaaD and yaaE genes. The pdxS (yaaD) mutant was confirmed to be a strict B(6) auxotroph, but the pdxT (yaaE) mutant turned out to be a conditional auxotroph depending on the availability of ammonium in the growth medium. The PdxS and PdxT proteins copurified during affinity chromatography and apparently form a complex that has glutaminase activity. PdxS and PdxT appear to encode the synthase and glutaminase subunits, respectively, of a glutamine amidotransferase of as-yet-unknown specificity essential for B(6) biosynthesis. PMID- 14762016 TI - PriA is essential for viability of the Escherichia coli topoisomerase IV parE10(Ts) mutant. AB - The parE10(Ts) mutation, which renders Escherichia coli thermosensitive for growth by inactivation of the essential E. coli topoisomerase topo IV, is lethal at all temperatures when PriA, the main replication restart protein, is absent. This lethality is suppressed by the activation of a PriA-independent replication restart pathway (dnaC809 mutation). This result suggests that topo IV acts prior to full-chromosome replication completion. PMID- 14762017 TI - Benzoate decreases the binding of cis,cis-muconate to the BenM regulator despite the synergistic effect of both compounds on transcriptional activation. AB - Fluorescence emission spectroscopy was used to investigate interactions between two effectors and BenM, a transcriptional regulator of benzoate catabolism. BenM had a higher affinity for cis,cis-muconate than for benzoate as the sole effector. However, the presence of benzoate increased the apparent dissociation constant (reduced the affinity) of the protein for cis,cis-muconate. Similar results were obtained with truncated BenM lacking the DNA-binding domain. High level transcriptional activation may require that some monomers within a BenM tetramer bind benzoate and others bind cis,cis-muconate. PMID- 14762018 TI - A temperature-sensitive mutation in the dnaE gene of Caulobacter crescentus that prevents initiation of DNA replication but not ongoing elongation of DNA. AB - A genetic screen for cell division cycle mutants of Caulobacter crescentus identified a temperature-sensitive DNA replication mutant. Genetic complementation experiments revealed a mutation within the dnaE gene, encoding the alpha-catalytic subunit of DNA polymerase III holoenzyme. Sequencing of the temperature-sensitive dnaE allele indicated a single base pair substitution resulting in a change from valine to glutamic acid within the C-terminal portion of the protein. This mutation lies in a region of the DnaE protein shown in Escherichia coli, to be important in interactions with other essential DNA replication proteins. Using DNA replication assays and fluorescence flow cytometry, we show that the observed block in DNA synthesis in the Caulobacter dnaE mutant strain occurs at the initiation stage of replication and that there is also a partial block of DNA elongation. PMID- 14762019 TI - Gamma-glutamyltranspeptidase, but not YwrD, is important in utilization of extracellular glutathione as a sulfur source in Bacillus subtilis. AB - gamma-Glutamyltranspeptidase (EC 2.3.2.2) of Bacillus subtilis, which is an extracellular enzyme, hydrolyzes the gamma-glutamyl linkage of glutathione. YwrD, which is homologous to gamma-glutamyltranspeptidase, was speculated to have a similar physiological role. It was shown that gamma-glutamyltranspeptidase, but not YwrD, is important in utilizing glutathione as the sole sulfur source in Bacillus subtilis. PMID- 14762020 TI - InvB is required for type III-dependent secretion of SopA in Salmonella enterica serovar Typhimurium. AB - The Salmonella effector protein SopA is translocated into host cells via the SPI 1 type III secretion system (TTSS) and contributes to enteric disease. We found that the chaperone InvB binds to SopA and slightly stabilizes it in the bacterial cytosol and that it is required for its transport via the SPI-1 TTSS. PMID- 14762021 TI - What now, mad cow? Experts put risk to US public in perspective. PMID- 14762022 TI - Ebola vaccines tested in humans, monkeys. PMID- 14762023 TI - Cancer vaccine research inches forward. PMID- 14762027 TI - Mechanisms and treatment of obstructive sleep apnea. PMID- 14762028 TI - Mechanisms and treatment of obstructive sleep apnea. PMID- 14762029 TI - Exogenous insulin and hypoglycemia as prognostic factors in critically ill patients. PMID- 14762030 TI - Exogenous insulin and hypoglycemia as prognostic factors in critically ill patients. PMID- 14762031 TI - Exogenous insulin and hypoglycemia as prognostic factors in critically ill patients. PMID- 14762032 TI - Sex, lies, and Niagra. PMID- 14762033 TI - Regulation of dietary supplements. PMID- 14762034 TI - Chemical composition of Chinese star anise (Illicium verum) and neurotoxicity in infants. PMID- 14762035 TI - Lowering homocysteine in patients with ischemic stroke to prevent recurrent stroke, myocardial infarction, and death: the Vitamin Intervention for Stroke Prevention (VISP) randomized controlled trial. AB - CONTEXT: In observational studies, elevated plasma total homocysteine levels have been positively associated with ischemic stroke risk. However the utility of homocysteine-lowering therapy to reduce that risk has not been confirmed by randomized trials. OBJECTIVE: To determine whether high doses of folic acid, pyridoxine (vitamin B6), and cobalamin (vitamin B12), given to lower total homocysteine levels, reduce the risk of recurrent stroke over a 2-year period compared with low doses of these vitamins. DESIGN: Double-blind randomized controlled trial (September 1996-May 2003). SETTING AND PARTICIPANTS: 3680 adults with nondisabling cerebral infarction at 56 university-affiliated hospitals, community hospitals, private neurology practices, and Veterans Affairs medical centers across the United States, Canada, and Scotland. INTERVENTIONS: All participants received best medical and surgical care plus a daily multivitamin containing the US Food and Drug Administration's reference daily intakes of other vitamins; patients were randomly assigned to receive once-daily doses of the high dose formulation (n = 1827), containing 25 mg of pyridoxine, 0.4 mg of cobalamin, and 2.5 mg of folic acid; or the low-dose formulation (n = 1853), containing 200 microg of pyridoxine, 6 microg of cobalamin and 20 microg of folic acid. MAIN OUTCOME MEASURES: Recurrent cerebral infarction (primary outcome); coronary heart disease (CHD) events and death (secondary outcomes). RESULTS: Mean reduction of total homocysteine was 2 micromol/L greater in the high-dose group than in the low-dose group, but there was no treatment effect on any end point. The unadjusted risk ratio for any stroke, CHD event, or death was 1.0 (95% confidence interval [CI], 0.8-1.1), with chances of an event within 2 years of 18.0% in the high-dose group and 18.6% in the low-dose group. The risk of ischemic stroke within 2 years was 9.2% for the high-dose and 8.8% for the low-dose groups (risk ratio, 1.0; 95% CI, 0.8-1.3) (P =.80 by log-rank test of the primary hypothesis of difference in ischemic stroke between treatment groups). There was a persistent and graded association between baseline total homocysteine level and outcomes. A 3- micromol/L lower total homocysteine level was associated with a 10% lower risk of stroke (P =.05), a 26% lower risk of CHD events (P<.001), and a 16% lower risk of death (P =.001) in the low-dose group and a nonsignificantly lower risk in the high-dose group by 2% for stroke, 7% for CHD events, and 7% for death. CONCLUSIONS: In this trial, moderate reduction of total homocysteine after nondisabling cerebral infarction had no effect on vascular outcomes during the 2 years of follow-up. However, the consistent findings of an association of total homocysteine with vascular risk suggests that further exploration of the hypothesis is warranted and longer trials in different populations with elevated total homocysteine may be necessary. PMID- 14762036 TI - Antiphospholipid antibodies and subsequent thrombo-occlusive events in patients with ischemic stroke. AB - CONTEXT: The presence of antiphospholipid antibodies (aPL) has been associated with vascular occlusive events. However, the role of aPL in predicting ischemic events, particularly recurrent ischemic stroke, is controversial. OBJECTIVE: To evaluate the effect of baseline aPL positivity (ie, positivity for anticardiolipin antibodies [aCL], lupus anticoagulant antibodies [LA], or both) on subsequent thrombo-occlusive events, including recurrent stroke. DESIGN, SETTING, AND PARTICIPANTS: The Antiphospholipid Antibodies and Stroke Study (APASS), a prospective cohort study within the Warfarin vs Aspirin Recurrent Stroke Study (WARSS), a randomized double-blind trial (N = 2206) conducted at multiple US clinical sites from June 1993 through June 2000 and comparing adjusted-dose warfarin (target international normalized ratio, 1.4-2.8) and aspirin (325 mg/d) for prevention of recurrent stroke or death. APASS participants were 1770 (80%) WARSS participants who consented to enroll in the APASS, with usable baseline blood samples drawn prior to randomization to the WARSS and analyzed for aPL status within 90 days of index stroke by a central independent laboratory. Quality assurance was performed on approximately 10% of samples by a second independent laboratory. MAIN OUTCOME MEASURE: Two-year rate of the composite end point of death from any cause, ischemic stroke, transient ischemic attack, myocardial infarction, deep vein thrombosis, pulmonary embolism, and other systemic thrombo-occlusive events. The primary analysis assessed the outcome associated with aPL positivity within each WARSS treatment group separately, after risk-factor adjustment (since these aPL-positive vs aPL negative comparisons were not randomized). RESULTS: Of the 1770 APASS patients, 720 (41%) were classified as aPL-positive and 1050 (59%) as aPL-negative. There was no increased risk of thrombo-occlusive events associated with baseline aPL status in patients treated with either warfarin (relative risk [RR], 0.99; 95% confidence interval [CI], 0.75-1.31; P =.94), or aspirin (RR, 0.94; 95% CI, 0.70 1.28; P =.71). The overall event rate was 22.2% among aPL-positive and 21.8% among aPL-negative patients. There was no treatment x aPL interaction (P =.91). Patients with baseline positivity for both LA and aCL antibodies tended to have a higher event rate (31.7%) than did patients who tested negative for both antibodies (24.0%) (unadjusted RR, 1.36; 95% CI, 0.97-1.92; P =.07). Classification and regression tree analyses did not identify a specific LA test or aCL isotype or titer that was associated with increased risk of thrombo occlusive event. CONCLUSIONS: The presence of aPL (either LA or aCL) among patients with ischemic stroke does not predict either increased risk for subsequent vascular occlusive events over 2 years or a differential response to aspirin or warfarin therapy. Routine screening for aPL in patients with ischemic stroke does not appear warranted. PMID- 14762037 TI - C-reactive protein and the risk of incident colorectal cancer. AB - CONTEXT: Inflammation may play a role in the pathogenesis of colorectal cancer; however, epidemiological evidence supporting this hypothesis in average-risk persons is sparse. OBJECTIVE: To determine the risk of incident colon and rectal cancer associated with elevated baseline plasma concentrations of C-reactive protein (CRP). DESIGN, SETTING, AND PARTICIPANTS: Prospective, nested case control study of a cohort of 22 887 adults (>18 years and Washington County, Maryland, residents) enrolled between May and October 1989 and followed up through December 2000. A total of 172 colorectal cancer cases were identified through linkage with the Washington County and Maryland State Cancer registries. Up to 2 controls (n = 342) were selected from the cohort for each case and matched by age, sex, race, and date of blood draw. MAIN OUTCOME MEASURE: Odds ratio (OR) of incident colon and rectal cancer. RESULTS: Plasma CRP concentrations were higher among all colorectal cases combined than controls (median CRP, 2.44 vs 1.94 mg/L; P =.01). The highest concentration was found in persons who subsequently developed colon cancer vs matched controls (median CRP, 2.69 vs 1.97 mg/L; P<.001). Among rectal cancer cases, CRP concentrations were not significantly different from controls (median CRP, 1.79 vs 1.81 mg/L; P =.32). The risk of colon cancer was higher in persons in the highest vs lowest quartile of CRP (OR, 2.55; 95% confidence interval [CI], 1.34-4.88; P for trend =.002). In nonsmokers, the corresponding association was stronger (OR, 3.51; 95% CI, 1.64-7.51; P for trend<.001). A 1-SD increase in log CRP (1.02 mg/L) was associated with an increased risk of colon cancer after adjusting for potential confounders and excluding cases occurring within 2 years of baseline (OR, 1.35; 95% CI, 1.05-1.74) or excluding those with late-stage colon cancer at the time of diagnosis (OR, 1.38; 95% CI, 0.99-1.91). CONCLUSIONS: Plasma CRP concentrations are elevated among persons who subsequently develop colon cancer. These data support the hypothesis that inflammation is a risk factor for the development of colon cancer in average-risk individuals. PMID- 14762038 TI - Resident-to-resident violent incidents in nursing homes. AB - CONTEXT: Little is known about nursing home residents' injuries that are inflicted by other residents. OBJECTIVE: To assess risk factors for violent injury to nursing home residents by other residents. DESIGN, SETTING, AND SUBJECTS: Case-control study using data from the Massachusetts Department of Public Health's Complaint and Incident Reporting System and from Minimum Data Set assessments for Massachusetts nursing home residents. Cases had an injury sustained from an incident with another nursing home resident between January 1, 2000, and December 31, 2000, which left visible evidence (ie, fracture, dislocation, bruise or hematoma, laceration, and reddened area) (median age, 81 years). Controls were randomly selected from all residents who had a Minimum Data Set assessment completed in 2000 (n = 101 429) and no injury report (median age, 83 years). A total of 1994 controls were included in the analyses. MAIN OUTCOME MEASURES: Injury type and risk of being injured by resident-to-resident aggressive physical behaviors based on the specific characteristics of the injured resident. RESULTS: During the first incident, 294 residents sustained fractures (n = 39), dislocations (n = 6), bruises or hematomas (n = 105), lacerations (n = 113), and reddened areas (n = 31). Injured residents (cases) were more likely to be cognitively impaired, exhibit symptoms of wandering, be verbally abusive, and have socially inappropriate behavior than the controls. Residents who were classified as needing extensive assistance (adjusted odds ratio [AOR], 0.3; 95% confidence interval [CI], 0.2-0.6) and being severely dependent (AOR, 0.12; 95% CI, 0.05-0.27) had a significant reduction in being injured. Residents in an Alzheimer disease unit were almost 3 times as likely to be injured than those living in other units (AOR, 3.2; 95% CI, 1.4-7.5). CONCLUSIONS: Injured residents were more likely, perhaps unknowingly, to "put themselves in harm's way," be verbally aggressive, and be cognitively impaired. Interventions to prevent these incidents should focus on the behavior of the injured persons. PMID- 14762039 TI - Ethical issues concerning research in complementary and alternative medicine. AB - The use of complementary and alternative medicine (CAM) has grown dramatically in recent years, as has research on the safety and efficacy of CAM treatments. Minimal attention, however, has been devoted to the ethical issues relating to research on CAM. We argue that public health and safety demand rigorous research evaluating CAM therapies, research on CAM should adhere to the same ethical requirements for all clinical research, and randomized, placebo-controlled clinical trials should be used for assessing the efficacy of CAM treatments whenever feasible and ethically justifiable. In addition, we explore the legitimacy of providing CAM and conventional therapies that have been demonstrated to be effective only by virtue of the placebo effect. PMID- 14762041 TI - The new antiepileptic drugs: clinical applications. AB - In the past decade, 8 new antiepileptic drugs have been approved for use in the United States, offering many new treatment options to patients with epilepsy. With expanding use of these newer agents, primary care clinicians are challenged with understanding the roles that each new agent plays in the treatment of patients with epilepsy as well as possible interactions with other pharmacological therapies. Each new medication provides a unique profile of pharmacokinetics, adverse effects, and mechanisms of action, making an appreciation of how these agents are best utilized even more difficult. Despite well-performed trials evaluating the safety and efficacy of specific antiepileptic drugs, the lack of head-to-head comparisons among them makes it difficult to endorse a single therapeutic regimen. Limited studies have compared the new antiepileptic drugs with more traditional medications and found similar efficacy but improved tolerability of the newer agents. There remains no well established guidelines for choosing a particular antiepileptic drug or for choosing a newer agent over a traditional one. However, careful consideration of seizure type, patient comorbidities, and specific medication toxicities aids in prescribing the most appropriate medication. This article aims to familiarize the general practitioner with the appropriate roles and effective uses of the new antiepileptic drugs in specific clinical scenarios. PMID- 14762040 TI - The new antiepileptic drugs: scientific review. AB - CONTEXT: The past decade has brought many advances to the treatment of epilepsy, including many new pharmacological agents. Primary care physicians often care for patients with epilepsy and therefore should be familiar with the new options available. OBJECTIVE: To review data regarding the efficacy and tolerability of antiepileptic drugs introduced in the past decade. DATA SOURCES: A search of the Cochrane Central Register of Controlled Trials was performed to identify all published human and English-language randomized controlled trials evaluating the efficacy and tolerability of the antiepileptic drugs that have been approved for use in the United States since 1990. Additional reports evaluating pharmacokinetic properties were identified through a MEDLINE search as well as review of article bibliographies. STUDY SELECTION AND DATA EXTRACTION: Search terms included felbamate, gabapentin, lamotrigine, topiramate, tiagabine, levetiracetam, oxcarbazepine, and zonisamide. Studies were selected if efficacy and tolerability were reported as major outcome measures. Included studies (n = 55) enrolled a minimum of 20 adult subjects and had a treatment period of at least 6 weeks. DATA SYNTHESIS: Eight new antiepileptic drugs have been approved for use in the United States in the past decade. Each new antiepileptic drug is well tolerated and demonstrates statistically significant reductions in seizure frequency over baseline. No randomized controlled trials have compared the new antiepileptic drugs with each other or against the traditional antiepileptic drugs. Although there is no evidence to suggest that the newer medications are more efficacious, several studies have demonstrated broader spectrum of activity, fewer drug interactions, and overall better tolerability of the new agents. CONCLUSIONS: New antiepileptic drugs offer many options in the treatment of epilepsy, each with unique mechanisms of action as well as adverse effect profiles. The new antiepileptic drugs are well tolerated with few adverse effects, minimal drug interactions, and a broad spectrum of activity. PMID- 14762042 TI - The challenge of stroke prevention. PMID- 14762043 TI - Is C-reactive protein an inflammation opsonin that signals colon cancer risk? PMID- 14762044 TI - STUDENTJAMA. Physician health and lifestyle. PMID- 14762045 TI - STUDENTJAMA. Physician burnout. PMID- 14762046 TI - STUDENTJAMA. Career satisfaction among physicians. PMID- 14762047 TI - STUDENTJAMA. Career satisfaction in female physicians. PMID- 14762048 TI - STUDENTJAMA. Residency training and pregnancy. PMID- 14762049 TI - STUDENTJAMA. Physician health and patient care. PMID- 14762050 TI - JAMA patient page. Epilepsy. PMID- 14762051 TI - Effect of srtA and srtB gene expression on the virulence of Staphylococcus aureus in animal models of infection. AB - OBJECTIVE: The role that the surface proteins anchored by the srtA and srtB gene products play in the ability of Staphylococcus aureus bacteria to establish infection was investigated in several animal models. METHODS: Wild-type and corresponding mutants with deletions of the srtA and/or srtB genes were used in murine acute lethal infection, septic arthritis, kidney infection and rat endocarditis models. RESULTS: The LD(50) of the wild-type and srtB- knockout were comparable and approximately two- to four-fold lower than the required inoculum of the srtA- and srtA-B- strains. This difference was exhibited as a two-fold greater mortality at the highest inoculum. The wild-type strain established arthritic inflammation in over 90% of the animals with a maximum arthritic index of 6.5 by days 17-21. The srtB- knockout was able to cause inflammation in 70-80% of the mice, but with a lower index of 3.0. Both the srtA- and srtA-B- strains appeared to be less virulent in this model with arthritic indices of around 0.5 and only 20% of the animals with inflammation. Strains with the srtA mutation achieved statistically significant lower titres than wild-type in kidneys of mice after intravenous infection. Mean bacterial counts in cardiac vegetations were significantly higher for the wild-type and srtB- strain compared with the srtA- and srtA-B- strains. CONCLUSION: Results from this study substantiate the role of the srtA gene product in the establishment of infections and further studies are warranted to define and exploit this as a target for antimicrobial chemotherapy. PMID- 14762052 TI - Survey of antibiotic knowledge amongst final year medical students. PMID- 14762053 TI - A vancomycin-resistant Enterococcus faecium isolate from a Danish healthy volunteer, detected 7 years after the ban of avoparcin, is possibly related to pig isolates. PMID- 14762054 TI - Methicillin-resistant Staphylococcus aureus bacteraemia diagnosed at hospital admission: distinguishing between community-acquired versus healthcare-associated strains. AB - BACKGROUND: Methicillin-resistant Staphylococcus aureus (MRSA) infections diagnosed at hospital admission are often referred to as community-acquired. This designation may include MRSA strains previously acquired in a healthcare setting (healthcare-associated) as well as those that have emerged from community-based S. aureus strains. METHODS: To understand further the epidemiology of MRSA from the community, a case-control study was performed. During 1997-2002, 254 patients with and without MRSA bacteraemia at hospital admission were studied. RESULTS: All patients with MRSA bacteraemia in the first 24 h of hospital admission had a recent exposure to a healthcare setting: true community-acquired MRSA was not detected. Independent risk factors for healthcare-associated MRSA bacteraemia, defined as MRSA bacteraemia in the first 24 h of hospital admission among patients with a recent exposure to a healthcare setting or intervention, included previous MRSA infection or colonization (OR = 17, P < 0.001), cellulitis (OR = 4, P = 0.006), presence of a central venous catheter (OR = 3, P < 0.001) and skin ulcers (OR = 3, P = 0.007). CONCLUSIONS: In this study, MRSA bacteraemia diagnosed in the first 24 h of hospital admission represented healthcare associated MRSA strains and not true community-acquired strains. The clinical characteristics associated with healthcare-associated MRSA bacteraemia can assist clinicians in targeting measures to prevent cross-transmission and may help to streamline empirical vancomycin therapy. PMID- 14762055 TI - Anti-staphylococcal activity and mode of action of clofazimine. AB - OBJECTIVES: Infections caused by Staphylococcus aureus might be treated with agents whose primary indications are for other infections. Clofazimine, an established anti-mycobacterial drug, could be such a candidate. However, the anti staphylococcal properties of clofazimine have not been fully described and its mode of action, possibly involving inhibition of both RNA polymerase and a membrane-located target, has not been explored in detail. We have now conducted experiments to address these issues. METHODS: Using established procedures, we examined the activity of clofazimine against a range of clinical isolates of S. aureus and determined whether it was bactericidal, exhibited a post-antibiotic effect (PAE), or interacted synergically with other agents. The potential for emergence of clofazimine-resistant mutants was also examined. Mode of action studies involved macromolecular synthesis assays, cross-screening against rifampicin-resistant mutants, susceptibility of RNA polymerase to clofazimine in vitro and several methods to detect drug-induced membrane damage. RESULTS: Clofazimine demonstrated good anti-staphylococcal activity encompassing MSSA, MRSA and GISA. It was bactericidal and resistant mutants could not be isolated. Clofazimine did not exhibit a PAE and failed to act synergically with other drugs. No evidence for specific inhibition of RNA polymerase was obtained. Clofazimine caused non-specific inhibition of DNA, RNA and protein synthesis, consistent with membrane-damaging activity that was detected in three independent assays for membrane disrupting agents. CONCLUSIONS: Clofazimine is a potent anti staphylococcal agent. It appears to be a membrane-disrupting agent and does not inhibit RNA polymerase. PMID- 14762056 TI - A randomized trial to investigate the recycling of stavudine and didanosine with and without hydroxyurea in salvage therapy (RESTART). AB - BACKGROUND: Treatment failure during highly active antiretroviral therapy (HAART) is ultimately common and associated with the development of resistance mutations to both the specific drug in question and cross-resistance to other available treatment options. In heavily pre-treated patients, the recycling of antiretroviral agents that have been utilized previously may, however, be associated with antiviral efficacy. We therefore conducted an investigation into the concept of recycling stavudine (d4T, Zerit) and didanosine (ddI, Videx) with and without hydroxyurea, in the management of heavily pre-treated HIV-1 infected individuals requiring salvage therapy (RESTART). METHODS: We randomized 21 individuals with treatment failure to receive stavudine and didanosine or stavudine, didanosine and hydroxyurea, for 12 weeks prior to optimizing therapy. Viral load, immunological parameters, genotypic information and the virtual phenotypes were obtained at baseline and at the end of the study. RESULTS: Significant decreases in viral loads were observed in both groups during a 12 week study period (P = 0.04), the addition of hydroxyurea conferring no additional benefit. This was not predicted by information from genotypes and virtual phenotypes, and these did not reveal sensitive or specific phenotypic cut offs for those individuals who responded to recycling. CONCLUSIONS: Salvage therapy with didanosine and stavudine can decrease viral loads in heavily pre treated individuals. Genotypic and virtual phenotype profiles provide little additional information in this setting. PMID- 14762057 TI - Effect of the efflux pump inhibitor Phe-Arg-beta-naphthylamide on the MIC values of the quinolones, tetracycline and chloramphenicol, in Escherichia coli isolates of different origin. PMID- 14762058 TI - A motif co-occurrence approach for genome-wide prediction of transcription-factor binding sites in Escherichia coli. AB - Various computational approaches have been developed for predicting cis regulatory DNA elements in prokaryotic genomes. We describe a novel method for predicting transcription-factor-binding sites in Escherichia coli. Our method takes advantage of the principle that transcription factors frequently coregulate gene expression, but without requiring prior knowledge of which groups of genes are coregulated. Using position weight matrices for 49 known transcription factors, we examined spacings between pairs of matrix hits. These pairs were assigned probabilities according to the overrepresentation of their separation distance. The functions of many open reading frames (ORFs) downstream from predicted binding sites are unknown, and may correspond to novel regulon members. For five predictions, knockouts with mutated replacements of the predicted binding sites were created in E. coli MG1655. Quantitative real-time PCR (RT-PCR) indicates that for each of the knockouts, at least one gene immediately downstream exhibits a statistically significant change in mRNA expression. This approach may be useful in analyzing binding sites in a variety of organisms. PMID- 14762060 TI - The genome sequence of Mycoplasma mycoides subsp. mycoides SC type strain PG1T, the causative agent of contagious bovine pleuropneumonia (CBPP). AB - Mycoplasma mycoides subsp. mycoidesSC (MmymySC)is the etiological agent of contagious bovine pleuropneumonia (CBPP), a highly contagious respiratory disease in cattle. The genome of Mmymy SC type strain PG1(T) has been sequenced to map all the genes and to facilitate further studies regarding the cell function of the organism and CBPP. The genome is characterized by a single circular chromosome of 1211703 bp with the lowest G+C content (24 mole%)and the highest density of insertion sequences (13% of the genome size)of all sequenced bacterial genomes. The genome contains 985 putative genes, of which 72 are part of insertion sequences and encode transposases. Anomalies in the GC-skew pattern and the presence of large repetitive sequences indicate a high genomic plasticity. A variety of potential virulence factors was identified, including genes encoding putative variable surface proteins and enzymes and transport proteins responsible for the production of hydrogen peroxide and the capsule, which is believed to have toxic effects on the animal. PMID- 14762061 TI - A comprehensive analysis of allelic methylation status of CpG islands on human chromosome 21q. AB - Approximately half of all human genes have CpG islands (CGIs)around their promoter regions. Although CGIs usually escape methylation, those on Chromosome X in females and those in the vicinity of imprinted genes are exceptions: They have both methylated and unmethylated alleles to display a "composite" pattern in methylation analysis. In addition, aberrant methylation of CGIs is known to often occur in cancer cells. Here we developed a simple HpaII-McrBC PCR method for discrimination of full, null, incomplete, and composite methylation patterns, and applied it to all computationally identified CGIs on human Chromosome 21q. This comprehensive analysis revealed that, although most CGIs (103 out of 149)escape methylation, a sizable fraction (31 out of 149)are fully methylated even in normal peripheral blood cells. Furthermore, we identified seven CGIs showing the composite methylation, and demonstrated that three of them are indeed methylated monoallelically. Further analyses using informative pedigrees revealed that two of the three are subject to maternal allele-specific methylation. Intriguingly, the other CGI is methylated in an allele-specific but parental-origin-independent manner. Thus, the cell seems to have a broader repertoire of methylating CGIs than previously thought, and our approach may contribute to uncover novel modes of allelic methylation. PMID- 14762062 TI - Evolutionary strata on the mouse X chromosome correspond to strata on the human X chromosome. AB - Lahn and Page previously observed that genes on the human X chromosome were physically arranged along the chromosome in "strata," roughly ordered by degree of divergence from related genes on the Y chromosome. They hypothesized that this ordering results from a historical series of suppressions of recombination along the mammalian Y chromosome, thereby allowing formerly recombining X and Y chromosomal genes to diverge independently. Here predictions of this hypothesis are confirmed in a nonprimate mammalian order, Rodentia, through an analysis of eight gene pairs from the X and Y chromosomes of the house mouse, Mus musculus. The mouse X chromosome has been rearranged relative to the human X, so strata were not found in the same physical order on the mouse X. However, based on synonymous evolutionary distances, X-linked genes in M. musculus fall into the same strata as orthologous genes in humans, as predicted. The boundary between strata 2 and 3 is statistically significant, but the boundary between strata 1 and 2 is not significant in mice. An analysis of smaller fragments of Smcy, Smcx, Zfy, and Zfx from seven species of Mus confirmed that the strata in Mus musculus were representative of the genus Mus. PMID- 14762063 TI - Patterns of evolutionary constraints in intronic and intergenic DNA of Drosophila. AB - We develop methods to infer levels of evolutionary constraints in the genome by comparing rates of nucleotide substitution in noncoding DNA with rates predicted from rates of synonymous site evolution in adjacent genes or other putatively neutrally evolving sites, while accounting for differences in base composition. We apply the methods to estimate levels of constraint in noncoding DNA of Drosophila. In introns, constraint (the estimated fraction of mutations that are selectively eliminated) is absolute at the 5' and 3' splice junction dinucleotides, and averages 72% in base pairs 3-6 at the 5'-end. Constraint at the 5' base pairs 3-6 is significantly lower in the lineage leading to Drosophila melanogaster than in Drosophila simulans, a finding that agrees with other features of genome evolution in Drosophila and indicates that the effect of selection on intron function has been weaker in the melanogaster lineage. Elsewhere in intron sequences, the rate of nucleotide substitution is significantly higher than at synonymous sites. By using intronic sites outside splice control regions as a putative neutrally evolving standard, constraint in the 500 bp of intergenic DNA upstream and downstream regions of protein-coding genes averages approximately 44%. Although the estimated level of constraint in intergenic regions close to genes is only about one-half of that of amino acid sites, selection against single-nucleotide mutations in intergenic DNA makes a substantial contribution to the mutation load in Drosophila. PMID- 14762064 TI - Mammalian overlapping genes: the comparative perspective. AB - It is believed that 3.2 billion bp of the human genome harbor approximately 35000 protein-coding genes. On average, one could expect one gene per 300000 nucleotides (nt). Although the distribution of the genes in the human genome is not random,it is rather surprising that a large number of genes overlap in the mammalian genomes. Thousands of overlapping genes were recently identified in the human and mouse genomes. However,the origin and evolution of overlapping genes are still unknown. We identified 1316 pairs of overlapping genes in humans and mice and studied their evolutionary patterns. It appears that these genes do not demonstrate greater than usual conservation. Studies of the gene structure and overlap pattern showed that only a small fraction of analyzed genes preserved exactly the same pattern in both organisms. PMID- 14762065 TI - High-resolution analysis of DNA copy number using oligonucleotide microarrays. AB - Genomic copy number alterations are a feature of many human diseases including cancer. We have evaluated the effectiveness of an oligonucleotide array, originally designed to detect single-nucleotide polymorphisms, to assess DNA copy number. We first showed that fluorescent signal from the oligonucleotide array varies in proportion to both decreases and increases in copy number. Subsequently we applied the system to a series of 20 cancer cell lines. All of the putative homozygous deletions (10) and high-level amplifications (12; putative copy number >4) tested were confirmed by PCR (either qPCR or normal PCR) analysis. Low-level copy number changes for two of the lines under analysis were compared with BAC array CGH; 77% (n = 44) of the autosomal chromosomes used in the comparison showed consistent patterns of LOH (loss of heterozygosity) and low-level amplification. Of the remaining 10 comparisons that were discordant, eight were caused by low SNP densities and failed in both lines. The studies demonstrate that combining the genotype and copy number analyses gives greater insight into the underlying genetic alterations in cancer cells with identification of complex events including loss and reduplication of loci. PMID- 14762066 TI - Digital detection of genetic mutations using SPC-sequencing. AB - Deletion or insertion mutations lead to a frameshift that causes misalignment between wild-type and mutated allele sequences, making it difficult to identify such mutations unambiguously by using electrophoresis-based DNA sequencing. We have previously established the feasibility of an accurate DNA sequencing method using solid-phase capturable (SPC) dideoxynucleotides and MALDI-TOF mass spectrometry on synthetic templates, an approach we refer to as SPC-sequencing. Here, we report the application of SPC-sequencing in characterizing frameshift mutations by using the detection of the BRCA1 gene mutations 185delAG and 5382insC as examples. In this method, Sanger DNA sequencing fragments are generated in one tube by using biotinylated dideoxynucleotides. The sequencing fragments carrying a biotin moiety at the 3' end are captured on a streptavidin coated solid phase to eliminate excess primer, primer dimers, and false stops. Only correctly terminated DNA fragments are captured, subsequently released, and analyzed by mass spectrometry to obtain digital DNA sequencing data. This method produces distinct doublet mass peaks at each point in the mass spectrum beyond the mutation site, facilitating the accurate characterization of the mutation. We have compared SPC-sequencing with electrophoresis-based sequencing in characterizing the above BRCA1 mutations, demonstrating the significant advantage offered by SPC-sequencing for the accurate identification of frameshift mutations. PMID- 14762067 TI - Chemical, physical, and sensorial characteristics of "Terrincho" ewe cheese: changes during ripening and intravarietal comparison. AB - The objectives of this study were to monitor the changes in chemical [moisture, acidity, pH, and water activity (a(w))] and physical (color and texture) parameters of "Terrincho" ewe cheese during 60 d of ripening, and to determine the correlations between the changes in instrumental texture and color parameters and the ripening time of the product. Intravarietal comparison of Terrincho ewe cheese from 5 different dairy plants was performed by evaluation of mechanical parameters from texture profile analysis (TPA) and color parameters in terms of CIELAB color space (L*, a*, and b*). In addition to mechanical and color tests, composition analyses and sensory tests were performed. The results were evaluated with statistical methods (single valued and multivariate analysis). During the first 20 d of ripening, an increase in hardness, fracturability, gumminess, chewiness, and yellowness occurred. Simultaneously, adhesiveness, resilience, L* (inside cheese, "i" and external "e"), and cohesiveness decreased. After 20 d of ripening hardness, fracturability, gumminess, and chewiness decreased and cohesiveness increased. The ripening time of Terrincho cheeses can be estimated with 6 variables: L* (external, e), L* (i), b* (inside cheese, i), hardness, a* (i), chewiness, and a constant. The estimation error was 4.2 d. Evaluation of composition, pH, texture profile analyses, color, and related sensory characteristics of Terrincho cheeses from 5 different dairy plants (with 30 d of ripening) revealed correlations between these parameters. PMID- 14762059 TI - Comparative genomics of gene expression in the parasitic and free-living nematodes Strongyloides stercoralis and Caenorhabditis elegans. AB - Although developmental timing of gene expression is used to infer potential gene function, studies have yet to correlate this information between species. We analyzed 10,921 ESTs in 3311 clusters from first- and infective third-stage larva (L1, L3i) of the parasitic nematode Strongyloides stercoralis and compared the results to Caenorhabditis elegans, a species that has an L3i-like dauer stage. In the comparison of S. stercoralis clusters with stage-specific expression to C. elegans homologs expressed in either dauer or nondauer stages, matches between S. stercoralis L1 and C. elegans nondauer-expressed genes dominated, suggesting conservation in the repertoire of genes expressed during growth in nutrient-rich conditions. For example, S. stercoralis collagen transcripts were abundant in L1 but not L3i, a pattern consistent with C. elegans collagens. Although a greater proportion of S. stercoralis L3i than L1 genes have homologs among the C. elegans dauer-specific transcripts, we did not uncover evidence of a robust conserved L3i/dauer 'expression signature.' Strikingly, in comparisons of S. stercoralis clusters to C. elegans homologs with RNAi knockouts, those with significant L1 specific expression were more than twice as likely as L3i-specific clusters to match genes with phenotypes. We also provide functional classifications of S. stercoralis clusters. PMID- 14762068 TI - Purification and characterization of three different types of bile salt hydrolases from Bifidobacterium strains. AB - Bile salt hydrolases were purified to electrophoretic homogeneity from Bifidobacterium bifidum ATCC 11863, Bifidobacterium infantis KL412, Bifidobacterium longum ATCC 15708, Bifidobacterium longum KL507, and Bifidobacterium longum KL515. Three different types (A, B, and C) of bile salt hydrolase (BSH) were revealed during the purification study, exhibiting the type specific characteristics in their electrophoretic migration and elution profiles from anion exchange and hydrophobic interaction chromatographic columns. The subunit molecular mass estimated by sodium dodecylsulfate (SDS)-polyacrylamide gel electrophoresis (PAGE) was around 35 kDa, and the native molecular mass in all five Bifidobacterium strains was estimated to be between 130 and 150 kDa by gel filtration chromatography, indicating that all BSH enzymes have tetrameric structure. From the isoelectric focusing, an isoelectric point value of 4.45 was obtained with BSH (type B) from B. bifidum ATCC 11863 and the other BSH (types A and C) showed the similar pI values around 4.65. N-Terminal amino acid sequencing for the proteins of types A and C revealed that 6 out of 20 amino acid residues were different, and highly conserved residues were identified in both N-terminal sequences of types A and C. All BSH enzymes from five strains hydrolyzed six major human bile salts, and they showed a better deconjugation rate on glycine conjugated bile salts than on taurine-conjugated forms. PMID- 14762069 TI - Effect of temperature and pore size on the fractionation of fresh and reconstituted buttermilk by microfiltration. AB - The objective of this research was to evaluate the effect of temperature (7, 25, and 50 degrees C) and pore size (0.1, 0.8, and 1.4 micro m) on the separation of proteins and lipids (neutral lipids and phospholipids) during microfiltration (MF) of fresh or reconstituted buttermilk. Buttermilk was subjected to MF using a pilot-scale unit mounted with ceramic membranes. The MF runs were carried out in a uniform transmembrane pressure (UTP) mode. Changes in processing temperature had no significant impact on protein transmission, whereas increasing temperature reduced both lipid and phospholipid transmission. A maximum concentration factor (CF) for lipids was reached at 25 degrees C, as protein CF remained essentially unaffected by temperature. The use of the smaller pore size (0.1 microm) resulted in low lipid (10%) and protein (approximately 20%) transmission. Larger pore sizes (0.8 and 1.4 microm) resulted in higher levels of protein, lipid, and phospholipid transmission (>50%), but gave high permeation fluxes. Transmission of both proteins and lipids was markedly different when using fresh buttermilk as opposed to reconstituted buttermilk. This study showed that MF temperature, pore size, and buttermilk type influence fractionation but that MF alone cannot achieve optimal separation of lipids and proteins for the production of novel ingredients from buttermilk. PMID- 14762070 TI - Combined use of chymosin and protease from Cryphonectria parasitica for control of meltability and firmness of cheddar cheese. AB - The combined use of chymosin and Cryphonectria parasitica protease was evaluated for manufacturing Cheddar cheese at different chymosin-to-C. parasitica protease ratios of 1:0, 0:1, 67:33, and 33:67. The degree of proteolysis over time was affected by the coagulant type. Proteolysis was thought to be the main cause of changes in functional properties of Cheddar cheeses. The meltability and hardness of cheese made with 100% C. parasitica enzyme was the highest, but it was high in bitterness. The chymosin-to-C. parasitica ratio between 0:1 to 67:33 was found suitable to independently control Cheddar cheese meltability and hardness without a significant level of bitterness. PMID- 14762071 TI - Components detected by means of solid-phase microextraction and gas chromatography/mass spectrometry in the headspace of artisan fresh goat cheese smoked by traditional methods. AB - The study of the headspace components of fresh smoked goat cheese, was carried out by means of solid-phase microextraction using a polyacrylate fiber followed by gas chromatography/mass spectrometry. The samples studied were six artisan Palmero cheeses manufactured following traditional methods and smoked using pine needles. The cheese regions studied were exterior, interior, and a cross section. In total, more than 320 components were detected, the exterior region being the richest in components, among which were acids, alcohols, esters, hydrocarbons, aldehydes, ketones, furan and pyran derivatives, terpenes and sesquiterpenes, nitrogen derivatives, phenol, guaiacol and syringol derivatives, ethers, and others. In addition to typical cheese components, typical smoke components were also detected; these latter were present especially in the headspace of the exterior region and only those in significant concentrations in the exterior region were also detected in the interior. The main components were acids and phenolic derivatives. These latter compounds play an important role in the flavor of this cheese, and their relative proportions together with the presence of specific smoke components derived from pine leaves may be considered of interest in order to distinguish this cheese from others smoked with different vegetable matter. PMID- 14762072 TI - The effect of Lactococcus lactis starter cultures on the oxidative stability of liquid whey. AB - The oxidative stability of liquid Cheddar cheese whey was evaluated using 2 Lactococcus lactis starter cultures in combination and alone along with a control, utilizing glucono-delta-lactone for acid development. Fresh and stored whey were evaluated for volatile composition, free fatty acids, and flavor by descriptive sensory analysis. A significant increase in volatile lipid oxidation products, most notably, hexanal, occurred during storage, and a corresponding decline in the free fatty acid linoleic acid was found. The flavor and aroma characteristic, cardboardy, was correlated to the increase in volatile lipid oxidation products and the decline in linoleic acid. Evidence strongly suggested that lipid oxidation was initiated during whey production and escalated during storage and that the starter cultures significantly influenced the level of volatile lipid oxidation products. Further understanding of the impact of starter cultures on whey may allow for the production of higher quality whey ingredients with wider food application. PMID- 14762073 TI - Contribution of native pasture to the sensory properties of Ragusano cheese. AB - Ragusano is a Protected Denomination of Origin cheese produced in the Hyblean area of Sicily. Sixteen samples of Ragusano cheese from two different treatments [pasture and total mixed ration (TMR)] were evaluated after 4 and 7 mo of aging. The color of the cheeses produced from milk of cows consuming fresh native pasture plants was much more yellow than cheeses from TMR fed cows (i.e., higher Hunter b value). This was due to transfer of beta-carotene and related compounds from the diet and demonstrated that compounds from native pasture plants changed the sensory characteristics of Ragusano cheese. To avoid a "halo" effect in a trained panel, quantitative descriptive analysis sensory evaluation of these cheeses for odor, taste, consistency, and mouth structure, color differences among cheeses were masked. A unique approach in sensory analysis was developed using sunglasses with lenses designed to block light at the specific wavelengths at which panelists would detect differences in color among samples. Testing was conducted every 2-wk period (15-d increments) with two tests per week using 11 trained panelists. All the panelists tasted all the products. Panelists were able to detect significant differences in the sensory characteristics of cheeses produced from milk of cows consuming native pastures versus TMR even when the color difference was masked. PMID- 14762074 TI - Growth responses of Escherichia coli to immunoglobulin G from cows immunized with ferric citrate receptor, FecA. AB - Effects of purified immunoglobulin (Ig) G from cows immunized with ferric citrate receptor, FecA, on the in vitro growth of Escherichia coli were investigated. Twenty-one cows were assigned to one of 3 treatments: 1) FecA immunization, 2) E. coli J5 bacterin immunization, and 3) unimmunized control. FecA was derived from E. coli UT5600/pSV66. Immunoglobulin G was purified from pooled colostral whey for each treatment group. The IgG from FecA immunized cows had higher titers against FecA compared with other treatment groups. Bacterial isolates tested were 14 E. coli from intramammary infections and E. coli UT5600/pSV66. Iron depletion decreased the growth of E. coli compared with growth in Fe-replete medium. The presence of IgG further decreased the growth compared with the growth under iron restriction alone. Bacterial growth did not differ among IgG sources nor between IgG concentrations. Replenishing media with exogenous iron overrode the inhibitory effects of the Fe-depletion and IgG. Vaccinating cows with FecA had little effect on the growth inhibitory properties of IgG toward E. coli mastitis isolates cultured in Fe-deplete media. PMID- 14762075 TI - Gamma-glutamyl transpeptidase inhibition suppresses milk protein synthesis in isolated ovine mammary cells. AB - The membrane spanning enzyme gamma-glutamyl transpeptidase (gamma-GT; EC 2.3.2.2) catalyses the breakdown of the tripeptide glutathione and uses free amino acids (AA) to form gamma-glutamyl transpeptidase (GT) AA that become transported into cells and converted back into free AA. gamma-Glutamyl transpeptidase activity has been shown to be important for mammary AA uptake in rodent systems, and while gamma-GT activity is high in lactating bovine mammary tissue, the role of this enzyme in milk protein synthesis of the ruminant has not been defined. The present study shows that gamma-GT activity in the ovine mammary gland, like that of rodents, increases during pregnancy and peaks early in lactation. Acivicin, a well-known inhibitor of gamma-GT, decreased gamma-GT activity in acini isolated from the ovine mammary gland and did not have secondary toxicity effects on cell viability or the uptake of radiolabeled amino-isobutyric acid. Isolated ovine acini were incubated in the presence of radiolabeled leucine, and incorporation of label into secreted protein increased during incubation. Incubation of acini with acivicin decreased milk protein secretion by 75%, indicating that gamma-GT plays an important role in milk protein production in the ruminant. Acivicin did not inhibit secretion of specific caseins but caused a global decrease in individual proteins suggesting that gamma-GT may be responsible for providing a complement of AA for milk protein synthesis. PMID- 14762076 TI - Risk of disease from wildlife reservoirs: badgers, cattle, and bovine tuberculosis. AB - Livestock face complex foraging options associated with optimizing nutrient intake while being able to avoid areas posing risk of parasites or disease. Areas of tall nutrient-rich swards around fecal deposits may be attractive for grazing, but might incur fitness costs from parasites. We use the example of dairy cattle and the risks of tuberculosis transmission posed to them by pastures contaminated with badger excreta to examine this trade-off. A risk may be posed either by aerosolized inhalation through investigation or by ingestion via grazing contaminated swards. We quantified the levels of investigation and grazing of 150 dairy cows at badger latrines (accumulations of feces and urine) and crossing points (urination-only sites). Grazing behavior was compared between strip-grazed and rotation-grazed fields. Strip grazing had fields subdivided for grazing periods of <24 h, whereas rotational grazing involved access to whole fields for 1 to 7 d each. A higher proportion of the herd investigated badger latrines than crossing points or controls. Cattle initially avoided swards around badger latrines but not around crossing points. Avoidance periods were shorter in strip compared with rotation-grazing systems. In rotation-grazing management, latrines were avoided for longer times, but there were more investigative contacts than with strip-grazing management. If investigation is a major route of tuberculosis transmission, the risk to cattle is greatest in extensive rotation-grazing systems. However, if ingestion of fresh urine is the primary method of transmission, strip-grazing management may pose a greater threat. Farming systems affect the level and type of contact between livestock and wildlife excreta and thus the risks of disease. PMID- 14762077 TI - Infusion of glucose directs circulating amino acids to the mammary gland in well fed dairy cows. AB - The effect of intestinal glucose supply on mammary utilization of amino acids (AA) was studied in four lactating dairy cows. Glucose (0, 443, 963, and 2398 g/d) was continuously infused in the duodenum over 14-d periods using a Latin square design. A grass silage-based diet was formulated so that treatments (diet + infusions) were isoenergetic and isonitrogenous and met 100 and 110% of energy and protein requirements, respectively. Mammary AA uptake was determined by arteriovenous difference and continuous blood flow measurement. The milk protein yield tended to be quadratically increased (to +88 g/d for 963 g of glucose) by glucose infusion, but milk protein content was not significantly affected. Treatments did not change significantly arterial concentrations of urea and glucogenic AA. Mammary arterial fluxes of essential AA increased linearly with glucose infusion, whereas fluxes of nonessential and glucogenic AA were not significantly affected. Mammary arteriovenous differences and extraction rates were roughly unchanged by treatments. Mammary uptake of all essential AA, excluding Arg and Val, increased linearly with increasing supply of glucose. Ratio of blood AA uptake to milk protein output increased significantly for His, Met, and Leu. For the highest infused dose of glucose, all AA except for His were taken up in excess relative to their secretion in milk. Based on evolution of extraction rate and ratio of uptake to output, His and Leu could have limited the milk protein yield response to glucose infusions. PMID- 14762078 TI - Hyperketonemia impairs glucose metabolism in pregnant and nonpregnant ewes. AB - The present study was undertaken to test the hypothesis that high ketone body concentrations suppress endogenous production of glucose and in pregnant sheep facilitate development of pregnancy toxemia. Rates of endogenous glucose production [mmol.min(-1)], and rate constants of glucose turnover [min(-1)] were measured in seven 12-h fasted sheep in the presence of normo- and hyperketonemia by use of D-2-[(3)H]-glucose. The measurements were carried out in the same sheep during the nonpregnant nonlactating state, during late pregnancy (10 +/- 7 d antepartum) and during lactation (19 +/- 6 d postpartum). Hyperketonemia (5 to 7 mmol.L(-1)), similar to that present in spontaneous ovine pregnancy toxemia, was induced by continuous intravenous 4-h infusions of DL-beta-hydroxybutyrate (DL BHB). Glucose turnover [mmol.min(-1)] in the same 7 nonpregnant nonlactating, late pregnant, and lactating sheep was significantly greater during normoketonemia (0.80, 1.16, 1.76) than during hyperketonemia (0.66, 0.92, 1.16, respectively). The rate constants of glucose turnover were not altered by elevation of the BHB concentration. The results demonstrated that high BHB concentrations significantly suppressed endogenous glucose production but showed no effect on glucose utilization. The suppressive effect of hyperketonemia on hepatic glucose production resulted in a significant reduction of plasma glucose concentration and was qualitatively the same in all three reproductive states. The results indicate that hyperketonemia, which is regularly present in late twin pregnant hypoglycemic sheep contributes significantly to the reduction of available glucose. This effect of hyperketonemia may invoke sustained hypoglycemia and may render the ewe into a vicious cycle that probably makes the animal refractory to treatment in most cases. PMID- 14762079 TI - Effect of low level monensin supplementation on the production of dairy cows fed alfalfa silage. AB - Effectiveness of low level monensin supplementation on N utilization in lactating dairy cows fed alfalfa silage was assessed using 48 multiparous Holsteins. Cows were fed a covariate diet [% of dry matter (DM): 56% alfalfa silage, 39% ground high moisture corn, 3% soybean meal, 1% ground corn, 1% vitamin-mineral supplements] for 2 wk, then grouped by days in milk into blocks of 4. Cows were randomly assigned within blocks to 1 of 4 diets that were fed for 10 wk: 1) control (covariate diet), 2) control plus 3% fish meal (replacing DM from high moisture corn), 3) monensin (10 mg/kg DM), and 4) monensin plus 3% fish meal. Diets 1 and 3 averaged 16.7% crude protein (25% from free AA in alfalfa silage); diets 2 and 4 averaged 18.5% crude protein. Monensin intake averaged 16 mg/d on diets 1 and 2 (due to contamination) and 248 mg/d on diets 3 and 4. There was no effect of fish meal or monensin on DM intake. However, weight gain and yield of milk, protein, and SNF increased with fish meal feeding, indicating metabolizable protein limited production. Feeding monensin increased blood glucose but reduced yield of 3.5% fat-corrected milk, milk fat content and yield, and milk protein content and yield. Apparent N efficiency was greatest on monensin (diet 3) but lowest on monensin plus fish meal (diet 4). Fish meal reduced blood glucose concentration and apparent N efficiency, and increased concentrations of milk and blood urea. Monensin increased ruminal propionate concentration and decreased concentration of acetate and butyrate and acetate:propionate in ruminally cannulated cows fed the experimental diets. However, these changes were small, suggesting that too little monensin was fed. Fish meal reduced ruminal total amino acid (AA) but monensin did not alter ruminal NH(3) or total AA. Both fish meal and monensin increased NH(3) formation from casein AA using ruminal inoculum from the cannulated cows. There was no evidence from this trial that feeding 250 mg of monensin per day to lactating cows improved N utilization by reducing ruminal catabolism of the large amounts of free AA in alfalfa silage. PMID- 14762080 TI - Endogenous synthesis of cis-9, trans-11 conjugated linoleic acid in dairy cows fed fresh pasture. AB - New Zealand Holstein-Friesian cows (n = 4) were used to quantify the importance of endogenous synthesis of cis-9, trans-11 conjugated linoleic acid (CLA) via Delta(9)-desaturase in cows fed a fresh pasture diet. The experiment was a 4 x 4 Latin square design with treatments arranged in a 2 x 2 factorial. Treatments lasted 4 d and were pasture only, pasture plus sterculic oil, pasture plus sunflower oil, and pasture plus sunflower oil plus sterculic oil. Abomasal infusion of sterculic oil inhibited Delta(9)-desaturase and decreased the concentration of cis-9, trans-11 CLA in milk fat by 70%. Using the changes in cis 9 10:1, cis-9 12:1 and cis-9 14:1 to correct for incomplete inhibition of Delta(9)-desaturase, a minimum estimate of 91% of cis-9, trans-11 CLA in milk fat was produced endogenously in cows fed fresh pasture. Dietary supplementation of a pasture diet with sunflower oil increased the proportion of long chain fatty acids in milk fat; however, the increase in vaccenic acid concentration was small (18%) and there was no increase in cis-9, trans-11 CLA concentration. Overall, results show that endogenous synthesis is responsible for more than 91% of the cis-9, trans-11 CLA secreted in milk fat of cows fed fresh pasture. However, the failure of plant oil supplements to increase the concentration of cis-9, trans-11 CLA in milk fat from pasture-fed cows requires further investigation. PMID- 14762081 TI - Apparent magnesium absorption in dry cows fed at 3 levels of potassium and 2 levels of magnesium intake. AB - In vitro experiments with isolated rumen epithelium have shown that the relationship between the ruminal K concentration and either the apical membrane potential difference or the mucosal-to-serosal Mg flux reach plateau values at high ruminal K concentrations. Hence, it may be hypothesized that the inhibitory effect of supplemental K on Mg absorption becomes smaller at high initial K intakes. To test our hypothesis, 6 ruminally fistulated, nonpregnant dry cows were fed 6 experimental diets in a 6 x 6 Latin square design with a 2 x 3 factorial arrangement of treatments. Four cows were of a Friesian-Holstein x Holstein-Friesian cross, and the 2 remaining cows were of a Meuse-Rhine-IJssel x Holstein-Friesian cross. The diets provided either 40.6 or 69.1 g of Mg per day and contained 20.7, 48.0, or 75.5 g of K per kilogram of dry matter. The dietary variables were obtained by mixing KHCO(3) and MgO into the basal concentrate. Absorption of Mg and the urinary Mg excretion was significantly decreased by supplemental K and significantly increased after the intake of supplemental Mg. In contrast to apparent Mg absorption, the urinary excretion of Mg was not affected by the dietary K x Mg interaction. Postfeeding ruminal K and Mg concentrations were increased with increasing K and Mg intakes. Postfeeding ruminal K concentrations and the urinary excretion of Mg showed a linear negative correlation; the slope was not significantly affected by Mg intake. Therefore, our hypothesis was rejected. Furthermore, these data indicate that supplemental Mg can effectively counteract the suppressant effect of K on Mg absorption in cows. PMID- 14762082 TI - Evaluation of milk urea nitrogen as a diagnostic of protein feeding. AB - An evaluation of milk urea nitrogen (MUN) as a diagnostic of protein feeding in dairy cows was performed using mean treatment data (n = 306) from 50 production trials conducted in Finland (n = 48) and Sweden (n = 2). Data were used to assess the effects of diet composition and certain animal characteristics on MUN and to derive relationships between MUN and the efficiency of N utilization for milk production and urinary N excretion. Relationships were developed using regression analysis based on either models of fixed factors or using mixed models that account for between-experiment variations. Dietary crude protein (CP) content was the best single predictor of MUN and accounted for proportionately 0.778 of total variance [MUN (mg/dL) = -14.2 + 0.17 x dietary CP content (g/kg dry matter)]. The proportion of variation explained by this relationship increased to 0.952 when a mixed model including the random effects of study was used, but both the intercept and slope remained unchanged. Use of rumen degradable CP concentration in excess of predicted requirements, or the ratio of dietary CP to metabolizable energy as single predictors, did not explain more of the variation in MUN (R(2) = 0.767 or 0.778, respectively) than dietary CP content. Inclusion of other dietary factors with dietary CP content in bivariate models resulted in only marginally better predictions of MUN (R(2) = 0.785 to 0.804). Closer relationships existed between MUN and dietary factors when nutrients (CP to metabolizable energy) were expressed as concentrations in the diet, rather than absolute intakes. Furthermore, both MUN and MUN secretion (g/d) provided more accurate predictions of urinary N excretion (R(2) = 0.787 and 0.835, respectively) than measurements of the efficiency of N utilization for milk production (R(2) = 0.769). It is concluded that dietary CP content is the most important nutritional factor influencing MUN, and that measurements of MUN can be utilized as a diagnostic of protein feeding in the dairy cow and used to predict urinary N excretion. PMID- 14762083 TI - Effect of supplemental L-lysine-HCL and corn source on rumen fermentation and amino acid flow to the small intestine. AB - Four lactating Jersey cows fitted with ruminal and duodenal cannulae were used in a 4 x 4 Latin square design trial to determine the effect of supplemental lysine in diets containing dry ground (GC) or steam-flaked (SFC, 360 g/L) corn on ruminal fermentation and amino acid (AA) flow to the duodenum. Supplemental L lysine-HCL provided 10 g/d of additional Lys to the total mixed rations. There were no interactions between supplemental Lys and corn source. Supplemental Lys increased Lys intake, but did not alter nutrient intake and digestibility or N flow to the duodenum. Intake of dry matter (DM), organic matter (OM), and neutral detergent fiber (NDF) and ruminal digestibility of starch tended to be higher, whereas ruminal digestibility of DM, OM, acid detergent fiber, and NDF was lower for diets supplemented with SFC compared with GC. Whole-tract digestibility was similar for both corn supplements. Ruminal pH and molar proportions of volatile fatty acids were not affected by supplemental Lys or corn source; however, ruminal NH(3) concentrations were lowest when SFC was fed. Intake of N tended to be higher and the flow of total N and individual AA to the duodenum was higher for diets supplemented with SFC. There was a trend for increased flow of microbial N for diets supplemented with SFC. Supplemental L-lysine-HCL did not alter ruminal fermentation, flow of amino acid to the small intestine, or nutrient digestibility, but feeding SFC reduced ruminal fiber digestion and increased microbial protein synthesis and flow of amino acid to the duodenum. PMID- 14762084 TI - Effects of silage species and supplemental vitamin E on the oxidative stability of milk. AB - Two experiments were conducted to study the effects of feeding legume silages and providing supplemental vitamin E in concentrates on the oxidative stability of milk. In experiment 1, six multiparous Holstein-Friesian cows were offered 1 of 6 silage treatments in a cyclical changeover-design experiment, with four 4-wk periods. The silages were grass, red clover, white clover, alfalfa, grass and red clover mixture (50:50 on a DM basis), and grass and white clover mixture (50:50 on a DM basis). In experiment 2, 8 cows were used in a changeover-design experiment with three 4-wk periods. The 4 treatments were a factorial combination of forages (grass silage or red clover silage) and supplemental vitamin E in the form of all-rac-alpha-tocopheryl acetate (29 or 290 IU/kg of DM in the concentrate). All forages were offered ad libitum and a flat rate of concentrates (8 kg/d) was fed in both experiments. Red clover silage led to significantly higher forage intakes, milk yields, and milk protein percentage in experiment 2, which was in agreement with results from experiment 1. There was no effect of vitamin E on feed intake, milk production, or milk fat and protein percentage. Red clover silage also led to significant changes in milk fatty acid profiles, particularly increased levels of polyunsaturated fatty acids. Milk samples were stored at 4 degrees C and 20 degrees C and analyzed for alpha-tocopherol and thiobarbituric acid reactive substances at intervals to determine oxidative stability. Diets based on red clover and alfalfa silages were associated with more rapid loss of alpha-tocopherol and increased production of thiobarbituric acid reactive substances during the storage of milk in comparison with diets based on grass silage. The increased oxidative deterioration of milk produced from cows fed red clover silage was avoided by vitamin E supplementation. PMID- 14762085 TI - Portal drained visceral flux, hepatic metabolism, and mammary uptake of free and peptide-bound amino acids and milk amino acid output in dairy cows fed diets containing corn grain steam flaked at 360 or steam rolled at 490 g/L. AB - Objectives were to measure net fluxes of free (FAA) and peptide bound amino acids (AA) (PBAA) across portal-drained viscera (PDV), liver, splanchnic, and mammary tissues, and of milk AA output of lactating Holstein cows (n = 6, 109 +/- 9 d in milk) as influenced by flaking density of corn grain. Cows were fed alfalfa-based total mixed ration (TMR) containing 40% steam-flaked (SFC) or steam-rolled corn (SRC) grain. The TMR were offered at 12-h intervals in a crossover design. Six sets of blood samples were obtained from indwelling catheters in portal, hepatic, and mammary veins and mesenteric or costoabdominal arteries every 2 h from each cow and diet. Intake of dry matter (18.4 +/- 0.4 kg/d), N, and net energy for lactation were not altered by corn processing. Milk and milk crude protein yields (kg/12-h sampling) were 14.2 vs. 13.5 and 0.43 vs. 0.39 for cows fed SFC or SRC, respectively. The PDV flux of total essential FAA was greater (571.2 vs. 366.4 g/12 h, SEM 51.4) in cows fed SFC. The PDV flux of total essential PBAA was 69.3 +/- 10.8 and 51.5 +/- 13.2 g/12 h for cows fed SFC and SRC, respectively, and differed from zero, but fluxes of individual PBAA rarely differed between treatments. Liver flux of essential FAA was greater in cows fed SRC, but only the PBAA flux in cows fed SRC differed from zero. Splanchnic flux of FAA and PBAA followed the pattern of PDV flux, but variation was greater. Mammary uptake (g/12 h) of total essential FAA was greater in cows fed SFC than SRC (224.6 vs. 198.3, SEM 7.03). Mammary uptake of essential PBAA was 25.0 vs. 15.1, SEM 5.2, g/12 h for cows fed SFC or SRC, respectively, and differed from zero in half of the PBAA. Milk output of EAA was 187.8 vs 175.4, SEM 4.4 g/12 h in cows fed SFC and SRC, respectively, and output of most essential AA consistently tended to be greater in cows fed SFC. It is apparent that PBAA comprise a portion of total AA flux across PDV and are affected by grain processing. Further, this pool supplies an important component of AA taken up by the mammary gland. Quantifying the contribution of PBAA may improve diet formulation with respect to intestinal absorption and mammary uptake of AA. PMID- 14762086 TI - The DGAT1 K232A mutation is not solely responsible for the milk production quantitative trait locus on the bovine chromosome 14. AB - The gene, acyl-CoA:diacylglycerol acyltransferase1 (DGAT1), was recently identified as the one underlying the quantitative trait locus (QTL) for milk production traits in the centromeric region of the bovine chromosome 14. Until now, 2 alleles, the lysine variant (increasing fat yield, fat and protein percentage) and the alanine variant (increasing protein and milk yield), were postulated at DGAT1. This study investigated whether the diallelic DGAT1 polymorphism is responsible for all the genetic variation at the centromeric region of this chromosome for milk, fat, and protein yield and fat and protein percentage. A statistical model was applied to a granddaughter design to analyze 16 German Holstein families. The model included the diallelic DGAT1 effect and the QTL transition probability estimated for each chromosomal position by a multiple marker approach. Because the regression coefficient of this probability was corrected for the diallelic DGAT1 polymorphism, it represented a putative conditional QTL effect. The effect of the DGAT1 gene was always highly significant. The conditional QTL effect was significant genomewise for fat percentage at the proximal end of the chromosome and for protein percentage at a more distal chromosomal region. Additional chromosomewise significance was found for fat and protein yield. Our results suggest an additional source of genetic variance on this chromosome for these traits; either one or more additional alleles segregating at DGAT1 that were not previously detected, a second quantitative trait locus affecting these traits, or both. PMID- 14762087 TI - Quantitative trait Loci for health traits in Finnish Ayrshire cattle. AB - A whole-genome scan was conducted to search for quantitative trait loci (QTL) affecting health traits in Finnish Ayrshire dairy cattle. The mapping population consisted of 12 bulls and their 491 sons in a granddaughter design. A total of 150 markers were typed covering all 29 autosomes. The traits under study were somatic cell score, mastitis, and a group of other veterinary treatments. Effects of the QTL and positions were estimated with the regression method. When carrying out interval mapping on each chromosome, cofactors were used to adjust for QTL identified at other chromosomes. Empirical P-values were obtained by permutation. Altogether 17 QTL were detected with genomewise significant P-values in the across family analysis. Quantitative trait loci affecting SCS were identified on chromosomes 1, 3, 11, 18, 21, 24, 27, 29, and QTL for mastitis on chromosomes 14, 18. Quantitative trait loci for other veterinary treatments were found on chromosomes 1, 2, 5, 8, 15, 22, and 23. The allele substitution effects were from 0.5 to 1.7 genetic standard deviations. The positions of these health QTL did not overlap with milk QTL detected in previous studies of the same population. PMID- 14762088 TI - Mapping quantitative trait loci affecting dairy conformation to chromosome 27 in two Holstein grandsire families. AB - Preliminary marker association results for quantitative trait loci affecting conformation traits using the granddaughter design and 8 large US Holstein grandsire families revealed strong associations in two families between the predicted transmitting abilities for dairy conformation and marker genotypes on bovine chromosome 27. Those results were based on single marker-trait associations in a genome-scan to identify broad chromosomal regions potentially containing genes affecting traits of interest. Results presented here describe continued study of quantitative trait loci on chromosome 27 for eventual incorporation into a marker-assisted selection program. Tests of marker associations for family 8 (91 sons) indicated an association with a microsatellite marker located near the telomere of chromosome 27. Interval analysis performed using additional marker genotypes generated for family 8 yielded further evidence for a quantitative trait locus in this region. No evidence was found for associations with milk production traits in this family in this region. An association was also detected in family 2 (240 sons) with a microsatellite marker located approximately 21 cM from the centromere of chromosome 27. Interval analysis performed for family 2 yielded evidence for a quantitative trait locus for dairy conformation near BMS689 with evidence of associations with fat percentage in the same region. Identification of quantitative trait loci affecting dairy conformation and fat components supports results reported by other groups, providing additional evidence that genes affecting fat metabolism are located on bovine chromosome 27. PMID- 14762089 TI - Genetic and phenotypic correlations between milk coagulation properties, milk production traits, somatic cell count, casein content, and pH of milk. AB - Genetic and phenotypic correlations between milk coagulation properties (MCP: coagulation time and curd firmness), milk yield, fat content, protein content, ln(somatic cell count) (SCS), casein content, and pH of milk and heritability of these traits were estimated from data consisting of milk samples of 4664 Finnish Ayrshire cows sired by 91 bulls. In addition, differences in average estimated breeding values (EBV) for the above traits between the cows with noncoagulating (NC) milk and those with milk that coagulated (CO samples) were examined. The estimations were carried out to study the possibilities of indirect genetic improvement of MCP by use of the above characteristics. The genetic and phenotypic correlations between MCP and the milk production traits were low or negligible. The genetic associations between desirable MCP and low SCS were rather strong (-0.45 to 0.29). Desirable MCP correlated both genetically and phenotypically with low pH of milk (-0.51 to 0.50). The rather high heritability estimates for curd firmness in different forms (0.22 to 0.39), and the wide variation in the proportion of daughters producing NC milk between the sires (0 to 47%) suggested that noncoagulation of milk is partly caused by additive genetic factors. Based on the genetic correlations between curd firmness and SCS and the high EBV for SCS obtained for the cows with NC-milk, it is possible that the loci causing noncoagulation of milk and increasing somatic cell count of milk are closely linked or partly the same. One means to genetically improve MCP and to reduce the occurrence of NC milk could thus be selection for low somatic cell count of milk. PMID- 14762090 TI - Detection of quantitative trait loci affecting milk production, health, and reproductive traits in Holstein cattle. AB - We report putative quantitative trait loci affecting female fertility and milk production traits using the merged data from two research groups that conducted independent genome scans in Dairy Bull DNA Repository grandsire families to identify quantitative trait loci (QTL) affecting economically important traits. Six families used by both groups had been genotyped for 367 microsatellite markers covering 2713.5 cM of the cattle genome (90%), with an average spacing of 7.4 cM. Phenotypic traits included PTA for pregnancy rate and daughter deviations for milk, protein and fat yields, protein and fat percentages, somatic cell score, and productive life. Analysis of the merged dataset identified putative quantitative trait loci that were not detected in the separate studies, and the pregnancy rate PTA estimates that recently became available allowed detection of pregnancy rate QTL for the first time. Sixty-one putative significant marker effects were identified within families, and 13 were identified across families. Highly significant effects were found on chromosome 3 affecting fat percentage and protein yield, on chromosome 6 affecting protein and fat percentages, on chromosome 14 affecting fat percentage, on chromosome 18 affecting pregnancy rate, and on chromosome 20 affecting protein percentage. Within-family analysis detected putative QTL associated with pregnancy rate on six chromosomes, with the effect on chromosome 18 being the most significant statistically. These findings may help identify the most useful markers available for QTL detection and, eventually, for marker-assisted selection for improvement of these economically important traits. PMID- 14762091 TI - A complete genome scan of the Israeli Holstein population for quantitative trait loci by a daughter design. AB - Eleven Israeli Holstein families including 5221 cows were analyzed by a daughter design for eight economic traits: milk, fat and protein production, fat and protein percentage, somatic cell score (SCS), herd-life, and female fertility. The cows were genotyped for 73 microsatellites with maximum spacing between markers of 53 cM. There were 86,304 informative genotypes. Preliminary analysis was by ANOVA of each trait, with the marker effect nested within sire. Significance was determined by controlling the false discovery rate at 0.4, after excluding markers with genome-wide significance for at least a single trait, and traits without any significant effects at this level. Thus, four markers on chromosomes 6 and 14 and female fertility were excluded. There remained 40 significant marker-trait combinations, and it is expected that 24 of these are true effects. To perform interval mapping for the families with significant contrasts, 21 additional markers were genotyped on chromosomes 2, 7, and 27. The bootstrap confidence intervals for gene effect did not include zero for protein percent on chromosome 2 and fat yield, protein yield, and SCS on chromosome 7. Quantitative trait locus heterozygosity was 33%, which is consistent with the hypothesis that only two alleles are segregating with unequal allele frequency. PMID- 14762092 TI - Quantitative trait loci mapping for dairy cattle production traits using a maximum likelihood method. AB - A maximum likelihood method was developed for QTL mapping in half-sib designs and compared to the regression method in analyses of both field and simulated data. The field data consisted of milk production evaluations of 433 progeny tested sons of 6 sires and 64 microsatellite markers distributed over 12 chromosomes. Based on permutation tests, 5 significant QTL were detected in the field data by the regression method compared with 10 by the maximum likelihood method (P < 0.05). In field data analysis, the maximum likelihood method detected more significant QTL and had a smaller residual variance than the regression method. The simulation included 9 scenarios differing in number of families, family size, QTL variance, and marker density, each replicated 100 times. The simulation results suggested that, as for the regression method, the precision of estimating QTL from the maximum likelihood method improves with increasing number of sons per sire, increasing the ratio of QTL to phenotypic variance, and decreasing marker interval. The maximum likelihood method had a smaller dispersion of estimated QTL positions than the regression method in 6 of 9 scenarios simulated. Overall, the maximum likelihood method shows potential advantage in QTL detection over the regression method, especially in the situations with less favorable conditions for QTL detection. PMID- 14762093 TI - Genotype x environment interaction for grazing versus confinement. I. Production traits. AB - The objective of this study was to investigate the possible existence of a genotype x environment interaction (GxE) for production traits of US Holsteins in grazing versus confinement herds. Grazing herds were defined as those that utilized grazing for at least 6 mo and were enrolled in dairy herd improvement (DHI). Control herds were confinement DHI herds of comparable size in similar regions. The performance of daughters in grazing herds and control herds was examined using linear regression of mature equivalent milk, fat, and protein yield on the November 2000 USDA-DHI predicted transmitting abilities (PTA) of their sires for those traits. Heritabilities and genetic correlations were estimated using restricted maximum likelihood in a bivariate animal model that considered the same trait in different environments as different traits. Product moment and rank correlations were calculated between sires' estimated breeding values, estimated separately in both environments. For grazing herds, the coefficient of regression of milk, fat and protein on PTA were 0.78, 0.76, and 0.78, respectively. Corresponding coefficients in the control herds were 0.99, 0.96, and 0.98. Estimates of heritability for the traits ranged from 0.2 to 0.25, and differences between grazing and control environments were small. Estimates of the genetic correlations for the traits in both environments were 0.89, 0.88, and 0.91 for milk, fat, and protein, respectively. Within-quartile analyses revealed a lower correlation for milk and protein between the upper and lower grazing quartiles, while the same quartiles for the control herds did not differ from unity. Rank correlation coefficients between sire estimated breeding values from the 2 environments were 0.59, 0.63, and 0.66 for milk, fat, and protein, respectively. The mean rank change for the top 100 sires between the two environments was 27. The regression coefficients indicate that expected daughter differences may be overstated by current sire PTA in grazing herds. Genetic correlations less than unity suggests that there is, at least, some reranking among sires in both environments, while the rank correlations indicate the possibility of sire reranking when evaluations were performed within management system. However, differences are not so large as to justify separate genetic evaluations for each system. PMID- 14762094 TI - Genotype x environment interaction for grazing vs. confinement. II. Health and reproduction traits. AB - Continual selection for increased milk yield for more than 40 yr, combined with the antagonistic relationship between increasing yield, somatic cell count, and fertility, have resulted in sires that may not be optimal for producing daughters for grazing systems where seasonal calving is very important. The objective of this study was to investigate the possible existence of a genotype x environment interaction (G x E) in grazing vs. confinement herds within the United States for lactation average somatic cell score (LSCS), days open (DO), days to first service (DFS), and number of services per conception (SPC). Grazing herds were defined as those that utilized grazing for at least 6 mo each year and were enrolled in Dairy Herd Improvement (DHI). Control herds were confinement DHI herds of similar size in the same states. For LSCS, the performance of daughters in grazing and control herds was examined using linear regression of LSCS on the November 2000 USDA-DHIA sire predicted transmitting abilities (PTA) for SCS. Genetic parameters for all traits were estimated using REML in a bivariate animal model that treated the same trait in different environments as different traits. Rank correlations were calculated between sires' estimated breeding values for LSCS, calculated separately for sires in both environments. The coefficient of regression of daughter LSCS on sire PTA was less in grazing herds than in control herds. The coefficient of regression for control herds was closer to expectation. Estimates of heritability were approximately 0.12 for LSCS, and less than 0.05 for the reproduction traits. Heritabilities for DO, DFS, and SPC were slightly higher for control herds. Estimates of genetic correlation for each reproductive trait between the 2 environments were high and not significantly different from unity. Generally, these traits appear to be under similar genetic control, but a lower coefficient of regression of LSCS on sire PTA for SCS in grazing herds suggests differences in daughter performance in grazing herds may be overstated based on current PTA for SCS. PMID- 14762095 TI - Effects of dry cow grouping strategy and prepartum body condition score on performance and health of transition dairy cows. AB - Multiparous Holstein cows (n = 337) on two commercial dairy farms were used to determine the effects of feeding a close-up diet for 21 (treatment S) or 60 d (treatment L). Milk yield was not affected by treatment; however, cows fed treatment S tended to have increased yields of fat, 3.5% fat-corrected milk, and protein during the first 5 mo of their subsequent lactation compared to treatment L. Cows fed treatment L gained more body condition score (BCS) during the dry period and had longer days to first service. As a secondary objective, relationships of BCS at dry off and subsequent performance were evaluated. Cows with initial BCS < or =3.0 (thinner) tended to produce more milk during early lactation than cows with initial BCS > or = 3.25 (fatter). A trend for an interaction of treatment and initial BCS existed for milk yield such that thinner cows fed treatment S produced the most milk and fatter cows fed treatment S produced the least amount of milk; cows fed treatment L regardless of BCS produced an intermediate amount of milk. Subsequent reproductive performance was similar among thinner and fatter cows. These data indicate that 2 group nutritional strategies for dry cows are preferred, and BCS at dry off should be considered when determining grouping and nutritional strategies for dry cows. Furthermore, moderately thin cows at dry off do not have impaired performance during their subsequent lactation compared to cows of greater BCS. PMID- 14762096 TI - Using activity and milk yield as predictors of fresh cow disorders. AB - The objective was to determine whether daily walking activity and milk yields could be used as predictors of metabolic and digestive disorders early in lactation. Data were collected from 1996 through 1999 from 1445 dairy cows in 3 Florida herds. Walking activity, milk yield, and other measures were collected from a computerized dairy management system. Mixed models analysis was used for data on cows before their first detected estrus, as identified by difference in activity. Healthy cows were defined as those without any metabolic or digestive disorder during the prebreeding stage, whereas a sick cow had an occurrence of those disorders at any time during the prebreeding stage. Metabolic disorders were ketosis, retained placenta, and milk fever. Digestive disorders included displaced abomasum, indigestion, reduced feed intake, traumatic gastritis, acidosis, and bloat. Data from cows with known cases of ketosis, left displaced abomasum, and digestive disorders were analyzed to determine changes in activity and milk yield before those specific disorders were clinically diagnosed. Although walking activity was generally lower among sick cows, cows with ketosis, left displaced abomasum, and digestive disorders had higher than average activity 8, 9, and 8 d, respectively, before each diagnosed disorder. Daily milk yields of sick cows were approximately 15 kg/d less than milk yields of healthy cows. Milk yields were lower by 6, 7, and 5 d, respectively, before diagnoses of ketosis, left displaced abomasum, and digestive disorders. Cows with ketosis, left displaced abomasum, and general digestive disorders could possibly be detected about 5 to 6 d earlier than clinical diagnoses based on changes in daily walking activity and milk yield. PMID- 14762097 TI - Protein kinase C epsilon and gamma: involvement in formalin-induced nociception in neonatal rats. AB - The central nervous system undergoes dynamic changes as it matures. However, until recently, very little was known about the impact of these changes on pain and analgesia. This study tested the hypothesis that the epsilon and gamma isozymes of protein kinase C (PKC) contribute to formalin-induced nociception in an age-dependent manner. Expression of epsilon and gamma PKC and the contributions of these isozymes in formalin-induced nociception was examined in postnatal day 7, 15, and 21 rats. epsilonPKC expression in dorsal root ganglion neurons and gammaPKC expression in lamina II of the spinal cord increased from the first to the third postnatal week. Coupling immunohistochemical and Western analysis, translocation of epsilonPKC followed intraplantar formalin in all ages. In contrast, formalin-induced gammaPKC translocation was observed only in postnatal day 21 rats. Behaviorally, intrathecal administration of the epsilonPKC specific inhibitor (epsilonV1-2) attenuated phase 1 and phase 2 formalin behaviors at all ages. In contrast, intrathecal administration of the gammaPKC specific inhibitor (gammaV5-3) attenuated only phase 2 responses in postnatal day 15 and 21 rats. Functionally, inhibition of epsilonPKC decreased capsaicin stimulated release of glutamate and calcitonin gene-related peptide in spinal cords isolated from postnatal day 7 rats. These results suggest that epsilonPKC age independently mediates inflammatory pain produced by intraplantar formalin. In contrast, gammaPKC contributes to formalin-induced nociception in an age dependent manner. Identifying the molecular mechanisms responsible for age specific patterns of nociception is necessary for the rational development of novel therapeutic strategies for treating pediatric pain. PMID- 14762098 TI - Enhancement of fibrinolysis by EF6265 [(S)-7-amino-2-[[[(R)-2-methyl-1-(3 phenylpropanoylamino)propyl]hydroxyphosphinoyl] methyl]heptanoic acid], a specific inhibitor of plasma carboxypeptidase B. AB - Plasma procarboxypeptidase B, also known as thrombin-activatable fibrinolysis inhibitor (TAFI), is converted by thrombin into the active enzyme, carboxypeptidase B (CPB)/activated TAFI. Plasma CPB down-regulates fibrinolysis by removing carboxy-terminal lysines, the ligands for plasminogen and tissue-type plasminogen activator (tPA), from partially degraded fibrin. To target thrombosis in a new way, we have identified and optimized a phosphinic acid-containing inhibitor of CPB, EF6265 [(S)-7-amino-2-[[[(R)-2-methyl-1-(3 phenylpropanoylamino) propyl]hydroxyphosphinoyl]methyl]heptanoic acid] and determined both the pharmacological profile and pathophysiological role of CPB in rat thrombolysis. EF6265 specifically inhibited plasma CPB activity with an IC(50) (50% inhibitory concentration) of 8.3 nM and enhanced tPA-mediated clot lysis in a concentration-dependent manner. EF6265 decreased detectable thrombi (percentage of glomerular fibrin deposition; control, 98 +/- 1.1; EF6265, 0.1 mg/kg, 27 +/- 9.1) that had been generated by tissue factor in a rat microthrombosis model with concomitant increases in plasma D-dimer concentration (control, <0.5 microg/ml; EF6265, 0.1 mg/kg, 15 +/- 3.5 microg/ml). EF6265 reduced plasma alpha2-antiplasmin activity to a lesser extent than tPA. In an arteriovenous shunt model, EF6265 (1 mg/kg) enhanced exogenous tPA-mediated thrombolysis under the same conditions that neither EF6265 nor tPA (600 kIU/kg) alone reduced thrombi. EF6265 (1 and 30 mg/kg) did not affect the bleeding time in rats. Moreover, it did not prolong the bleeding time evoked by tPA (600 kIU/kg). These results confirm that circulating procarboxypeptidase B functions as a fibrinolysis inhibitor's zymogen and validates the use of CPB inhibitors as both an enhancer of physiological fibrinolysis in microcirculation and as a novel adjunctive agent to tPA for thromboembolic diseases while maintaining a small effect on primary hemostasis. PMID- 14762099 TI - Mouse reduced in osteosclerosis transporter functions as an organic anion transporter 3 and is localized at abluminal membrane of blood-brain barrier. AB - The "reduced in osteosclerosis" transporter (Roct), which shows decreased expression in the osteosclerosis (oc) mutant mouse, has high homology with rat and human organic anion transporter 3 (OAT3). However, its transport properties and involvement in bone turnover are poorly understood. Here, we examined Roct mediated transport using a Xenopus laevis oocyte expression system. Roct expressing oocytes exhibited uptake of [(3)H]estrone sulfate, [(3)H]p aminohippuric acid, [(3)H]benzylpenicillin, [(3)H]estradiol 17beta-glucronide, [(3)H]indoxyl sulfate, [(14)C]indomethacin, [(3)H]homovanillic acid, [(3)H]cimetidine, [(14)C]glutarate, [(14)C]salicylic acid, and [(3)H]methotrexate. Furthermore, the uptake of [(3)H]benzylpenicillin by Roct coexpressed with Na(+)-dicarboxylate cotransporter was trans-stimulated by glutarate preloading, and [(3)H]estrone sulfate uptake showed a similar tendency, suggesting that Roct is a dicarboxylate exchanger. [(3)H]Benzylpenicillin uptake by Roct was inhibited by OAT3 substrates and inhibitors, and by sulfate or glucuronide conjugates, and compounds involved in bone turnover. Roct mRNA is expressed abundantly in the kidney and was also detected in the brain, choroid plexus, and eye. Immunohistochemical analysis revealed that Roct is localized in brain capillary endothelial cells. These results indicate that the transport properties and tissue distribution of Roct are similar to those of OAT3, suggesting that Roct functions as mouse OAT3. Because Roct is expressed in the kidney and at the blood-brain barrier, it may play a role in the excretion of substrates such as conjugates and bone turnover factors. PMID- 14762100 TI - Cooperation between aspirin-triggered lipoxin and nitric oxide (NO) mediates antiadhesive properties of 2-(Acetyloxy)benzoic acid 3-(nitrooxymethyl)phenyl ester (NCX-4016) (NO-aspirin) on neutrophil-endothelial cell adherence. AB - 2-(Acetyloxy)benzoic acid 3-(nitrooxymethyl)phenyl ester (NCX-4016) is a nitric oxide (NO)-releasing derivative of aspirin that inhibits cyclooxygenase (COX) activity and releases NO. Acetylation of COX-2 by aspirin activates a transcellular biosynthetic pathway that switches eicosanoid biosynthesis from prostaglandin E(2) to 15-epi-lipoxin (LX)A(4) or aspirin-triggered lipoxin (ATL). Here, we demonstrate that exposure of neutrophil (PMN)/human umbilical vein endothelial cell (HUVEC) cocultures to aspirin and NCX-4016 triggers ATL formation and inhibits cell-to-cell adhesion induced by endotoxin (LPS) and interleukin (IL)-1beta by 70 to 90%. However, although selective and nonselective COX-2 inhibitors (celecoxib, rofecoxib, and naproxen) or N-t butoxycarbonylmethionine-leucine-phenylalanine (Boc-1), an LXA(4) receptor antagonist, reduced the antiadhesive properties of aspirin by approximately 70%, antiadhesive effects of NCX-4016 were only marginally affected ( approximately 30%) by COX inhibitors and Boc-1, implying that COX-independent mechanisms mediate the antiadhesive properties of NCX-4016. Indeed, NCX-4016 causes a long lasting (up to 12 h) release of NO and cGMP accumulation in HUVEC. Scavenging NO with 10 mM hemoglobin, in the presence of celecoxib, reduced the antiadhesive properties of NCX-4016 by approximately 80%. Confirming a role for NO, the NO donor diethylenetriamine-NO also inhibited PMN/HUVEC adhesion by approximately 80%. NCX-4016, but not aspirin, decreased DNA binding of nuclear factor-kappaB (NF-kappaB) on gel shift analysis and HUVEC's overexpression of CD54 and CD62E induced by LPS/IL-1beta. Reduction of binding of the two NF-kappaB subunits p50 p50 and p50-p65 was reversed by dithiothreitol, implying S-nitrosylation as mechanism of inhibition. In summary, our results support that ATL and NO are formed at the PMN/HUVEC interface after exposure to NCX-4016 and mediate the antiadhesive properties of this compound. PMID- 14762101 TI - Ethanol antagonist peptides: structural specificity without stereospecificity. AB - Increasing evidence suggests that ethanol damages the developing nervous system partly by disrupting the L1 cell adhesion molecule. Ethanol inhibits L1-mediated cell adhesion, and compounds that antagonize this action also prevent ethanol induced embryotoxicity. Two such compounds are the small peptides NAPVSIPQ (NAP) and SALLRSIPA (SAL). We showed previously that NAP and SAL antagonize ethanol inhibition of L1 adhesion at femtomolar to picomolar concentrations. Here we demonstrate that, despite this extraordinary potency, both NAP and SAL lack stereospecificity. d-NAP, a peptide composed entirely of d-amino acids, was an effective ethanol antagonist in NIH/3T3 cells transfected with human L1 and in the NG108-15 neural cell line. Interestingly, Ala-substituted derivatives of d NAP demonstrate the same structure-activity relation as the corresponding derivatives of l-NAP. The Ser-Ile-Pro motif was important for the ethanol antagonist activity of d-NAP, l-NAP, and l-SAL, with Ile being the most critical element in all three. Like l-NAP, d-NAP effectively reduced ethanol-induced growth retardation in mouse whole embryo culture. The potential resistance of d peptides to proteases makes d-NAP a potentially attractive agent for the prevention of fetal alcohol syndrome. PMID- 14762102 TI - Multidrug resistance protein (MRP) 4- and MRP 5-mediated efflux of 9-(2 phosphonylmethoxyethyl)adenine by microglia. AB - The pathogenesis of human immunodeficiency virus (HIV)-associated dementia has been linked to microglial responses after infection. We have recently confirmed expression of several ATP-dependent efflux transporters in microglia, namely, multidrug resistance protein 1 (MRP1) and P-glycoprotein (P-gp). In the present study, we investigated whether cultured rat microglia express two additional MRP family members, rMRP4 and rMRP5. Using reverse transcriptase-polymerase chain reaction, rMRP4 and rMRP5 mRNA was detected in primary cultures of microglia and in a rat microglia cell line, MLS-9. Western blot analysis further confirmed protein expression of the two MRP isoforms in MLS-9 cells. Bis(pivaloxymethyl)-9 (2-phosphonylmethoxyethyl)adenine [bis(POM)PMEA], a lipophilic ester prodrug of the well characterized MRP4 and 5 substrate 9-(2-phosphonylmethoxyethyl)adenine (PMEA), was chosen to examine transport characteristics in MLS-9. Using thin layer chromatography, we verified that more than 90% of radioactivity recovered in MLS-9 loaded with 1 microM [(3)H]bis(POM)PMEA for 1 h under ATP-depleting conditions was converted to PMEA. Efflux of PMEA by MLS-9 cell monolayers was ATP dependent, glutathione-independent, and significantly inhibited by several MRP inhibitors (i.e., sulfinpyrazone, genistein, indomethacin, and probenecid) as well as the antiretroviral drug azidothymidine-monophosphate. Similar results were not observed in MRP1- or P-gp-overexpressing cell lines, suggesting that PMEA is not a substrate for either P-gp or MRP1. These studies provide further evidence that microglia express multiple subfamilies of ATP-binding cassette transporters (i.e., P-gp, MRP1, MRP4, and MRP5) that could restrict permeation of several different classes of antiretroviral drugs in a brain cellular target of HIV-1 infection. PMID- 14762103 TI - Ethanol causes inflammation in the airways by a neurogenic and TRPV1-dependent mechanism. AB - Ethanol (EtOH) stimulates peptidergic primary sensory neurons via the activation of the transient receptor potential vanilloid-1 (TRPV1). EtOH is also known to trigger attacks of asthma in susceptible individuals. Our aim was to investigate whether EtOH produces airway inflammation via a TRPV1-dependent mechanism and to verify whether this effect is produced via a mechanism distinct from that of acetaldehyde (AcH). EtOH caused a Ca(2+)-dependent release of neuropeptides from guinea pigs airways, an effect that was inhibited by both capsaicin pretreatment and the TRPV1 antagonist capsazepine (CPZ). Furthermore, EtOH contracted isolated guinea pig bronchi, showing efficacy similar to that of carbachol: this effect of EtOH was sensitive to capsaicin pretreatment, tachykinin receptor blockade, and TRPV1 antagonism. The EtOH metabolite AcH also contracted isolated guinea pig bronchi, but this action was not affected by capsaicin pretreatment, tachykinin receptor, or TRPV1 antagonism. EtOH by intravenous or intragastric route of administration caused bronchoconstriction and increased plasma extravasation in the guinea pig airways, effects that were abolished selectively by CPZ. In conclusion, we have demonstrated that EtOH stimulates peptidergic primary sensory neurons in the guinea pig airways by TRPV1 activation. This excitatory effect of EtOH, distinct from that of AcH, results in neurogenic inflammatory responses that may contribute to the mechanism of EtOH-induced asthma. PMID- 14762104 TI - Structural determinants of the pharmacological properties of the GABAA receptor alpha6 subunit. AB - GABA(A) receptors are responsible for fast inhibitory neurotransmission in the mammalian brain and are the targets for many clinical drugs that act as antiepileptics, anxiolytics, and sedatives. The pharmacological characteristics of the receptor are largely determined by its subunit composition. Compared with all other alpha subtypes, the alpha6 subtype confers unique pharmacological properties. In particular, alpha6-containing receptors are more sensitive to both the agonist GABA and the antagonist amiloride. Results from chimeric constructs of the alpha1 and alpha6 subunits suggested that structural differences within the extracellular N-terminal domain were responsible for both these characteristics. Within this domain, we examined 15 amino acid residues unique to the alpha6 subtype. Each of these sites was individually mutated in the alpha6 subunit to the corresponding residue of the alpha1 subunit. The mutated subunits were expressed in human embryonic kidney-293T cells along with wild-type beta3 and gamma2L subunits and sensitivity to GABA and amiloride determined with whole cell electrophysiological recordings. Serine83 in the alpha6 subunit influenced sensitivity to both GABA and amiloride. Leucine174 and tyrosine175 were also found to contribute to inhibition by amiloride but did not affect GABA sensitivity. These structural differences are at least partly responsible for the unique pharmacological properties associated with the alpha6 subunit. PMID- 14762105 TI - Cdk5 regulates axonal transport and phosphorylation of neurofilaments in cultured neurons. AB - Phosphorylation has long been considered to regulate neurofilament (NF) interaction and axonal transport, and, in turn, to influence axonal stability and their maturation to large-caliber axons. Cdk5, a serine/threonine kinase homologous to the mitotic cyclin-dependent kinases, phosphorylates NF subunits in intact cells. In this study, we used two different haptenized NF subunits and manipulated cdk5 activity by microinjection, transfection and pharmacological inhibition to monitor the effect of Cdk5-p35 on NF dynamics and transport. We demonstrate that overexpression of cdk5 increases NF phosphorylation and inhibits NF axonal transport, whereas inhibition both reduces NF phosphorylation and enhances NF axonal transport in cultured chicken dorsal-root-ganglion neurons. Large phosphorylated-NF 'bundles' were prominent in perikarya following cdk5 overexpression. These findings suggest that Cdk5-p35 activity regulates normal NF distribution and that overexpression of Cdk5-p35 induces perikaryal accumulation of phosphorylated-NFs similar to those observed under pathological conditions. PMID- 14762106 TI - Specific in vivo phosphorylation sites determine the assembly dynamics of vimentin intermediate filaments. AB - Intermediate filaments (IFs) continuously exchange between a small, depolymerized fraction of IF protein and fully polymerized IFs. To elucidate the possible role of phosphorylation in regulating this equilibrium, we disrupted the exchange of phosphate groups by specific inhibition of dephosphorylation and by specific phosphorylation and site-directed mutagenesis of two of the major in vivo phosphorylation sites determined in this study. Inhibition of type-1 (PP1) and type-2A (PP2A) protein phosphatases in BHK-21 fibroblasts with calyculin-A, induced rapid vimentin phosphorylation in concert with disassembly of the IF polymers into soluble tetrameric vimentin oligomers. This oligomeric composition corresponded to the oligopeptides released by cAMP-dependent kinase (PKA) following in vitro phosphorylation. Characterization of the (32)P-labeled vimentin phosphopeptides, demonstrated Ser-4, Ser-6, Ser-7, Ser-8, Ser-9, Ser-38, Ser-41, Ser-71, Ser-72, Ser-418, Ser-429, Thr-456, and Ser-457 as significant in vivo phosphorylation sites. A number of the interphase-specific high turnover sites were shown to be in vitro phosphorylation sites for PKA and protein kinase C (PKC). The effect of presence or absence of phosphate groups on individual subunits was followed in vivo by microinjecting PKA-phosphorylated (primarily S38 and S72) and mutant vimentin (S38:A, S72:A), respectively. The PKA-phosphorylated vimentin showed a clearly decelerated filament formation in vivo, whereas obstruction of phosphorylation at these sites by site-directed mutagenesis had no significant effect on the incorporation rates of subunits into assembled polymers. Taken together, our results suggest that elevated phosphorylation regulates IF assembly in vivo by changing the equilibrium constant of subunit exchange towards a higher off-rate. PMID- 14762107 TI - The surface of articular cartilage contains a progenitor cell population. AB - It is becoming increasingly apparent that articular cartilage growth is achieved by apposition from the articular surface. For such a mechanism to occur, a population of stem/progenitor cells must reside within the articular cartilage to provide transit amplifying progeny for growth. Here, we report on the isolation of an articular cartilage progenitor cell from the surface zone of articular cartilage using differential adhesion to fibronectin. This population of cells exhibits high affinity for fibronectin, possesses a high colony-forming efficiency and expresses the cell fate selector gene Notch 1. Inhibition of Notch signalling abolishes colony forming ability whilst activated Notch rescues this inhibition. The progenitor population also exhibits phenotypic plasticity in its differentiation pathway in an embryonic chick tracking system, such that chondroprogenitors can engraft into a variety of connective tissue types including bone, tendon and perimysium. The identification of a chondrocyte subpopulation with progenitor-like characteristics will allow for advances in our understanding of both cartilage growth and maintenance as well as provide novel solutions to articular cartilage repair. PMID- 14762108 TI - The N-myristoylated Rab-GTPase m-Rabmc is involved in post-Golgi trafficking events to the lytic vacuole in plant cells. AB - We report on the sub-cellular localisation and function of m-Rab(mc), a N myristoylated plant-specific Rab-GTPase previously characterised at the molecular level and also by structural analysis in Mesembryanthemum crystallinum. By confocal laser scanning microscopy, we identified m-Rab(mc) predominantly on the prevacuolar compartment of the lytic vacuole but also on the Golgi apparatus in various plant cell types. Two complementary approaches were used immunocytochemistry and cyan fluorescent protein (CFP)/yellow fluorescent protein (YFP)-fusion proteins. Co-localisation studies of m-Rab(mc) with a salinity stress modulated integral calcium-ATPase suggest involvement of m-Rab(mc) in a plant-specific transport pathway to the prevacuolar compartment of the lytic vacuole. This hypothesis was strengthened by the inhibition of the transport of aleurain fused to green fluorescent protein (GFP), a marker of the lytic vacuole, in the presence of the dominant negative mutant m-Rab(mc)(N147I) in Arabidopsis thaliana protoplasts. The inhibitory effect of m-Rab(mc)(N147I) was specific for the transport pathway to the lytic vacuole, since the transport of chitinase-YFP, a marker for the neutral vacuole, was not hindered by the mutant. PMID- 14762109 TI - Cascade pathway of filopodia formation downstream of SCAR. AB - The protrusion of two distinct actin-containing organelles, lamellipodia and filopodia, is thought to be regulated by two parallel pathways: from Rac1 through Scar/WAVEs to lamellipodia, and from Cdc42 through N-WASP to filopodia. We tested this hypothesis in Drosophila, which contains a single gene for each WASP subfamilies, SCAR and WASp. We performed targeted depletion of SCAR or WASp by dsRNA-mediated interference in two Drosophila cultured cell lines expressing lamellipodial and filopodial protrusion. Knockdown was verified by laser capture microdissection and RT-PCR, as well as western blotting. Morphometrical, kinetic and electron microscopy analyses of the SCAR-depleted phenotype in both cell types revealed strong inhibition of lamellipodial formation and cell spreading, as expected. More importantly, filopodia formation was also strongly inhibited, which is not consistent with the parallel pathway hypothesis. By contrast, depletion of WASp did not produce any significant phenotype, except for a slight inhibition of spreading, showing that both lamellipodia and filopodia in Drosophila cells are regulated predominantly by SCAR. We propose a new, cascade pathway model of filopodia regulation in which SCAR signals to lamellipodia and then filopodia arise from lamellipodia in response to additional signal(s). PMID- 14762110 TI - Overexpression of latent transforming growth factor-beta binding protein 1 (LTBP 1) in dioxin receptor-null mouse embryo fibroblasts. AB - The aryl hydrocarbon receptor (AhR) is a transcriptional regulator of genes involved in xenobiotic metabolism. Increasingly clear is also the role of the AhR in the control of cell growth and proliferation. By analyzing differential patterns of gene expression between wild-type (AhR+/+) and null (AhR-/-) mouse embryo fibroblasts (MEF), we have identified latent transforming growth factor beta binding protein 1 (LTBP-1) as a negatively AhR-regulated gene in the absence of xenobiotics. Ltbp-1 mRNA and protein expression were markedly increased in AhR /- MEF. Furthermore, secreted LTBP-1 was elevated in the culture medium and the extracellular matrix of AhR-null MEF. Actinomycin D inhibited Ltbp-1 mRNA overexpression, suggesting regulation at the transcriptional level. AhR activation by dioxin (TCDD) downregulated Ltbp-1, again suggesting an AhR regulated mechanism. Treatment of AhR+/+ MEF with transforming growth factor beta(TGF-beta) downregulated AhR and, simultaneously, increased Ltbp-1, further supporting the role of this receptor in LTBP-1 expression. AhR-/- conditioned medium had higher levels of active and total TGF-beta activity, suggesting a role for LTBP-1 in maintaining extracellular TGF-beta concentrations. TGF-beta did not appear to directly regulate Ltbp-1 given that addition of TGFbeta neutralizing antibody or TGFbeta protein to AhR-/- MEF had no effect on Ltbp-1 expression. AhR /- MEF had lower levels of matrix metalloproteinase 2 (MMP-2) activity, which could not be attributable to MMP-2 mRNA downregulation or MMP-inhibitors Timp-1 and Timp-2 overexpression. These data identify LTBP-1 as one of the few AhR regulated genes not involved in xenobiotic metabolism and also support the implication of the AhR in controlling TGFbeta activity and cell proliferation. PMID- 14762112 TI - Endocytic trafficking in actively resorbing osteoclasts. AB - Endocytosis and the subsequent intracellular trafficking of the endocytosed material are important determinants of cellular function. Osteoclasts, cells of the monocyte/macrophage family, are specialized for the internalization and processing of bone matrix. Transcytosis of endocytosed material has been observed in osteoclasts but the precise mechanism controlling this process is unclear. Here, we investigate the regulation of these trafficking events. To establish the directionality and kinetics of trafficking events in resorbing osteoclasts, we devised a system using fluorescent low-molecular-weight markers as probes to follow the route taken by the digested bone matrix. We demonstrate that this route is largely distinct from the pathway followed by proteins taken up by receptor-mediated endocytosis at the basolateral plasma membrane. Endocytosis and transcytosis from the ruffled border are fast processes, with a half-life of the endocytosed material inside the cells of 22 minutes. We demonstrate the crucial role of the microtubule network in transport from the ruffled-border area and provide evidence for a role of the cytoskeleton in the overall efficacy of trafficking. Moreover, we analyse the effect of the V-ATPase inhibitor bafilomycin A1 on endocytic uptake, which gives insight into the pH-dependent regulation of membrane trafficking and resorption in osteoclasts. PMID- 14762111 TI - Molecular cross-talk between the transcription, translation, and nonsense mediated decay machineries. AB - It is widely believed that translation occurs only in the cytoplasm of eukaryotes, but recent results suggest some takes place in nuclei, coupled to transcription. Support for this heterodoxy comes from studies of the nonsense mediated decay (NMD) pathway; this pathway probably uses ribosomes to proofread messenger RNAs. We find components of the machineries involved in transcription, translation and NMD colocalise, interact and copurify, and that interactions between them are probably mediated by the C-terminal domain of the catalytic subunit of RNA polymerase II. These results are simply explained if the NMD machinery uses nuclear ribosomes to translate - and so proofread - newly made transcripts; then, faulty transcripts and any truncated peptides produced by nuclear translation would be degraded. PMID- 14762113 TI - The amount of neurofilaments aggregated in the cell body is controlled by their increased sensitivity to trypsin-like proteases. AB - Neurofilaments are synthesised and assembled in neuronal cell bodies, transported along axons and degraded at the synapse. However, in several pathological situations they aggregate in cell bodies or axons. To investigate their turnover when separated from their normal site of degradation, we used a previously described transgenic model characterised by perikaryal retention of neurofilaments, and compared the basic features of both neurofilament synthesis and degradation with that observed in normal mice. Despite the massive perikaryal aggregates, neurofilament transcript levels were found to be unchanged, whereas the total accumulation of neurofilament proteins was markedly reduced. Neurofilaments isolated from transgenic samples are more sensitive to both trypsin and alpha-chymotrypsin mediated proteolysis. Consistent with their greater in vitro sensitivity, trypsin immunolabeling of cell bodies was stronger in transgenic mice. These results show a novel mechanism to regulate the amount of neurofilaments when they abnormally aggregate. PMID- 14762114 TI - Evidence for structural and functional diversity among SDS-resistant SNARE complexes in neuroendocrine cells. AB - The core complex, formed by the SNARE proteins synaptobrevin 2, syntaxin 1 and SNAP-25, is an important component of the synaptic fusion machinery and shows remarkable in vitro stability, as exemplified by its SDS-resistance. In western blots, antibodies against one of these SNARE proteins reveal the existence of not only an SDS-resistant ternary complex but also as many as five bands between 60 and >200 kDa. Structural conformation as well as possible functions of these various complexes remained elusive. In western blots of protein extracts from PC12 cell membranes, an antibody against SNAP-25 detected two heat-sensitive SDS resistant bands with apparent molecular weights of 100 and 230 kDa. A syntaxin antibody recognized only the 230 kDa band and required heat-treatment of the blotting membrane to detect the 100 kDa band. Various antibodies against synaptobrevin failed to detect SNARE complexes in conventional western blots and detected either the 100 kDa band or the 230 kDa band on heat-treated blotting membranes. When PC12 cells were exposed to various extracellular K(+) concentrations (to evoke depolarization-induced Ca(2+) influx) or permeabilized in the presence of basal or elevated free Ca(2+), levels of these SNARE complexes were altered differentially: moderate Ca(2+) rises (anterior (preferred) direction with mRNAs from the retina stimulated in the anterior-->posterior (null) direction. Acyl-CoA-binding protein (ACBP) mRNA was one of four mRNAs selected by this screen, the proteins of which interact with GABA receptors. HOKS in the preferred direction increased ACBP mRNA transcription and ACBP protein expression. ACBP was localized to Muller glial cells by hybridization histochemistry and by immunohistochemistry. ACBP interacts with the alpha1 subunit of the GABA(A) receptor, as determined by a yeast two-hybrid technique. This interaction was confirmed by coimmunoprecipitation of ACBP and the alpha1 subunit of the GABA(A) receptor using an antibody to GABA(A)alpha1. The interaction was also confirmed by a "pull-down" assay in which histidine-tagged ACBP was used to pull down the GABA(A)alpha1. ACBP does not cross the blood-brain barrier. However, smaller truncated proteolytic fragments of ACBP do, increasing the excitability of central cortical neurons. Muller cells may secrete ACBP in the inner plexiform layer, thereby decreasing the sensitivity of GABA(A) receptors expressed on the surface of ganglion cell dendrites. Because retinal directional sensitivity is linked to GABAergic transmission, HOKS-induced expression of ACBP could provide a molecular basis for adaptation to HOKS and for the genesis of OKAN-II. PMID- 14762121 TI - A new rat model of the human serial reaction time task: contrasting effects of caudate and hippocampal lesions. AB - There is often little correspondence between human and animal examples of nondeclarative memory. The serial reaction time task (SRT) is a sequence learning example of human nondeclarative memory that may be suitable for development as an animal model. The SRT is believed to be impaired by basal ganglia, not limbic system damage, but there is uncertainty whether limbic system pathology does in fact leave the SRT unimpaired. We therefore developed a new rat model that closely approximated the human SRT, using intracranial self-stimulation to promote rapid continuous responding to four adjacent nose pokes in a single test session. Intact rats that experienced repeated sequences demonstrated robust interference effects when switched to a random sequence of cued responses (at 4-, 8-, and 12-sequence lengths), unlike intact controls that experienced the random sequences only. The interference effect in the human task is a key measure for nondeclarative sequence learning. Rats with dorsal caudate lesions that experienced massed sequence repetitions showed an interference effect at the four sequence length only. By contrast, rats with dorsal hippocampal lesions showed an interference effect at all sequence lengths. This new rat SRT model clarifies the basal ganglia-limbic system dichotomy suggested by human work. PMID- 14762122 TI - Dopaminergic modulation of prefrontal cortical input to nucleus accumbens neurons in vivo. AB - Dopaminergic transmission in the nucleus accumbens has been proposed to modulate the effects of converging excitatory inputs from the cortex, hippocampus, and amygdala. Here, we used in vivo intracellular recording in anesthetized rats to examine the response of nucleus accumbens neurons to stimulation of the prefrontal cortex (PFC) and the ventral tegmental area (VTA). The EPSP elicited in accumbens neurons by PFC stimulation was attenuated by VTA train stimulation in a pattern mimicking dopamine cell burst firing. PFC-elicited EPSPs were smaller in amplitude and faster to decay after VTA stimulation. These changes could not be explained by membrane depolarization alone, because EPSPs evoked during the sustained depolarization after VTA stimulation were significantly smaller than EPSPs evoked during spontaneously occurring up states. Furthermore, no attenuation of PFC-elicited responses was observed during depolarization produced by positive current injection through the recording electrode. Administration of a D1 antagonist (SCH 23390; 0.5 mg/kg, i.p.) had no effect on the VTA reduction of PFC-elicited responses, whereas administration of a D2 antagonist (eticlopride; 0.5 mg/kg, i.p.) reversed the reduction of PFC inputs when the analysis was limited to comparisons with spontaneous up states. These results suggest that the ability of PFC inputs to drive accumbens neurons is dampened by dopamine acting primarily at D2 receptors. Along with previous reports of dopaminergic attenuation of limbic afferents to the accumbens, these findings support the hypothesis that dopamine mediates the selection and integration of excitatory inputs and thus shapes information processing in accumbens output neurons. PMID- 14762123 TI - Intracranial self-administration of ethanol within the ventral tegmental area of male Wistar rats: evidence for involvement of dopamine neurons. AB - Previous work from our laboratory indicated that female Wistar rats will self administer ethanol (EtOH) directly into the posterior ventral tegmental area (VTA). These results suggested that VTA dopamine (DA) neurons might be involved in mediating the reinforcing actions of EtOH within this region. The objectives of this study were to determine (1) the dose-response effects for the self administration of EtOH into the VTA of male Wistar rats, and (2) the involvement of VTA DA neurons in the reinforcing actions of EtOH within the VTA. Adult male Wistar rats were implanted stereotaxically with guide cannulas aimed at the posterior or anterior VTA. After 1 week, rats were placed in standard two-lever (active and inactive) experimental chambers for a total of seven to eight sessions. The first experiment determined the intracranial self-administration of EtOH (0-400 mg%) into the posterior and anterior VTA. The second experiment examined the effects of coadministration of the D2/3 agonist quinpirole on the acquisition and maintenance of EtOH self-infusions into the posterior VTA. The final experiment determined the effects of a D2 antagonist (sulpiride) to reinstate self-administration behavior in rats given EtOH and quinpirole to coadminister. Male Wistar rats self-infused 100-300 mg% EtOH directly into the posterior, but not anterior, VTA. Coadministration of quinpirole prevented the acquisition and extinguished the maintenance of EtOH self-infusion into the posterior VTA, and addition of sulpiride reinstated EtOH self-administration. The results of this study indicate that EtOH is reinforcing within the posterior VTA of male Wistar rats and suggest that activation of VTA DA neurons is involved in this process. PMID- 14762124 TI - Ventral pallidal representation of pavlovian cues and reward: population and rate codes. AB - We recorded neural activity in the ventral pallidum (VP) while rats learned a pavlovian reward association. Rats learned to distinguish a tone that predicted sucrose pellets (CS+) from a different tone that predicted nothing (CS-). Many VP units became responsive to CS+, but few units responded to CS-. When two CS+ were encountered sequentially, the earliest predictor of reward became most potent. Many VP units were also activated when the sucrose reward was received [unconditioned stimulus (UCS)]. These VP units for UCS remained responsive to sucrose reward after learning, even when sucrose was already predicted by CS+. Neural representation of reward learning and reward itself was characterized by population codes. The population of units that responded to CS+ increased with learning, whereas the population that responded to UCS did not change. A relative firing rate code also represented the identities of conditioned stimuli and UCS. Firing rate differences among stimuli were acquired early and remained stable during subsequent training, whereas population codes and behavioral conditioned responses continued to develop during subsequent training. Thus, the VP makes use of dynamic CS population and rate codes to encode pavlovian reward cues in reward learning and uses stable UCS population and firing codes to encode sucrose reward itself. PMID- 14762125 TI - Ephrin-A5 exerts positive or inhibitory effects on distinct subsets of EphA4 positive motor neurons. AB - Eph receptor tyrosine kinases and ephrins are required for axon patterning and plasticity in the developing nervous system. Typically, Eph-ephrin interactions promote inhibitory events; for example, prohibiting the entry of neural cells into certain embryonic territories. Here, we show that distinct subsets of motor neurons that express EphA4 respond differently to ephrin-A5. EphA4-positive LMC(l) axons avoid entering ephrin-A5-positive hindlimb mesoderm. In contrast, EphA4-positive MMC(m) axons extend through ephrin-A5-positive rostral half sclerotome. Blocking EphA4 activation in MMC(m) neurons or expanding the domain of ephrin-A5 expression in the somite results in the aberrant growth of MMC(m) axons into the caudal half-sclerotome. Moreover, premature expression of EphA4 in MMC(m) neurons leads to a portion of their axons growing into novel ephrin-A5 positive territories. Together, these results indicate that EphA4-ephrin-A5 signaling acts in a positive manner to constrain MMC(m) axons to the rostral half sclerotome. Furthermore, we show that Eph activation localizes to distinct subcellular compartments of LMC(l) and MMC(m) neurons, consistent with distinct EphA4 signaling cascades in these neuronal subpopulations. PMID- 14762126 TI - L and M cone contributions to the midget and parasol ganglion cell receptive fields of macaque monkey retina. AB - Analysis of cone inputs to primate parvocellular ganglion cells suggests that red green spectral opponency results when connections segregate input from long wavelength (L) or middle wavelength (M) sensitive cones to receptive field centers and surrounds. However, selective circuitry is not an obvious retinal feature. Rather, cone receptive field surrounds and H1 horizontal cells get mixed L and M cone input, likely indiscriminately sampled from the randomly arranged cones of the photoreceptor mosaic. Red-green spectral opponency is consistent with random connections in central retina where the mixed cone ganglion cell surround is opposed by a single cone input to the receptive field center, but not in peripheral retina where centers get multiple cone inputs. The selective and random connection hypotheses might be reconciled if cone type selective circuitry existed in inner retina. If so, the segregation of L and M cone inputs to receptive field centers and surrounds would increase from horizontal to ganglion cell, and opponency would remain strong in peripheral retina. We measured the relative strengths of L and M cone inputs to H1 horizontal cells and parasol and midget ganglion cells by recording intracellular physiological responses from morphologically identified neurons in an in vitro preparation of the macaque monkey retina. The relative strength of L and M cone inputs to H1 and ganglion cells at the same locations matched closely. Peripheral midget cells were nonopponent. These results suggest that peripheral H1 and ganglion cells inherit their L and M cone inputs from the photoreceptor mosaic unmodified by selective circuitry. PMID- 14762127 TI - Linearity of cortical receptive fields measured with natural sounds. AB - How do cortical neurons represent the acoustic environment? This question is often addressed by probing with simple stimuli such as clicks or tone pips. Such stimuli have the advantage of yielding easily interpreted answers, but have the disadvantage that they may fail to uncover complex or higher-order neuronal response properties. Here, we adopt an alternative approach, probing neuronal responses with complex acoustic stimuli, including animal vocalizations. We used in vivo whole-cell methods in the rat auditory cortex to record subthreshold membrane potential fluctuations elicited by these stimuli. Most neurons responded robustly and reliably to the complex stimuli in our ensemble. Using regularization techniques, we estimated the linear component, the spectrotemporal receptive field (STRF), of the transformation from the sound (as represented by its time-varying spectrogram) to the membrane potential of the neuron. We find that the STRF has a rich dynamical structure, including excitatory regions positioned in general accord with the prediction of the classical tuning curve. However, whereas the STRF successfully predicts the responses to some of the natural stimuli, it surprisingly fails completely to predict the responses to others; on average, only 11% of the response power could be predicted by the STRF. Therefore, most of the response of the neuron cannot be predicted by the linear component, although the response is deterministically related to the stimulus. Analysis of the systematic errors of the STRF model shows that this failure cannot be attributed to simple nonlinearities such as adaptation to mean intensity, rectification, or saturation. Rather, the highly nonlinear response properties of auditory cortical neurons must be attributable to nonlinear interactions between sound frequencies and time-varying properties of the neural encoder. PMID- 14762128 TI - Nuclear factor E2-related factor 2-dependent antioxidant response element activation by tert-butylhydroquinone and sulforaphane occurring preferentially in astrocytes conditions neurons against oxidative insult. AB - Binding of the transcription factor nuclear factor E2-related factor 2 (Nrf2) to the antioxidant response element (ARE) in neural cells results in the induction of a battery of genes that can coordinate a protective response against a variety of oxidative stressors. In this study, tert-butylhydroquinone (tBHQ) and sulforaphane were used as activators of this pathway. Consistent with previous studies, treatment of primary cortical cultures from ARE reporter mice revealed selective promoter activity in astrocytes. This activation protected neurons from hydrogen peroxide and nonexcitotoxic glutamate toxicity. tBHQ treatment of cultures from Nrf2 knock-out animals resulted in neither ARE activation nor neuroprotection. By reintroducing Nrf2 via infection with a replication-deficient adenovirus (ad), both the genetic response and neuroprotection were rescued. Conversely, infection with adenovirus encoding dominant-negative (DN) Nrf2 (ad-DN Nrf2) or pretreatment with the selective phosphatidylinositol-3 kinase inhibitor LY294002 inhibited the tBHQ-mediated promoter response and corresponding neuroprotection. Interestingly, the adenoviral infection showed a high selectivity for astrocytes over neurons. In an attempt to reveal some of the cell type-specific changes resulting from ARE activation, cultures were infected with adenovirus encoding green fluorescent protein (GFP) (ad-GFP) or ad-DN-Nrf2 (containing GFP) before tBHQ treatment. A glia-enriched population of GFP infected cells was then isolated from a population of uninfected neurons using cell-sorting technology. Microarray analysis was used to evaluate potential glial versus neuron-specific contributions to the neuroprotective effects of ARE activation and Nrf2 dependence. Strikingly, the change in neuronal gene expression after tBHQ treatment was dependent on Nrf2 activity in the astrocytes. This suggests that Nrf2-dependent genetic changes alter neuron-glia interactions resulting in neuroprotection. PMID- 14762129 TI - Fast remapping of sensory stimuli onto motor actions on the basis of contextual modulation. AB - Higher organisms can establish complex associations between sensory events and motor responses. More remarkable than their complexity, however, is that the resulting sensory-motor maps can be selectively interchanged. For example, a person who speaks English and Spanish can read aloud "con once, sin once," going effortlessly from one language to the other. What is the neural basis of this capacity? Here, a network model is presented in which multiple maps between sensory stimuli and motor actions are possible, but only one of them, depending on behavioral context, is implemented at any given time. The key is a nonlinear representation in which the gain of sensory responses is regulated by context information. Neuronal responses can indeed show variations in gain, as has been documented in the case of proprioceptive signals such as eye and head position, which can modulate visually triggered activity. However, in contrast to these, the contextual cues used here need not bear any relationship to the physical attributes of the stimuli; in particular, spatial location is irrelevant. The model thus postulates the existence of sensory neurons that are nonlinearly modulated by arbitrary context signals, a plausible and testable prediction. The proposed mechanism allows a network of neurons to effectively change the functional connectivity between its inputs and outputs and may partially explain how animals can quickly adapt their behavior to varying environmental conditions. PMID- 14762130 TI - Intracellular association of glycine receptor with gephyrin increases its plasma membrane accumulation rate. AB - Gephyrin, a tubulin-binding protein, is the core of inhibitory postsynaptic scaffolds stabilizing glycine receptors (GlyRs) and/or GABA(A) receptors. Previous ultrastructural studies in vivo and in vitro have reported a localization of gephyrin to intracellular cisternas during development or after glycinergic denervation (Seitanidou et al., 1992; Colin et al., 1996, 1998). These data were compatible with a traffic of this cytoplasmic, but membrane associated, protein together with membrane proteins such as GlyR after exocytosis and/or endocytosis pathways. We have now investigated the consequences of a GlyR gephyrin interaction on the localization and the dynamics of these two molecules in African green monkey kidney cells (COS-7) cells and in neurons transfected with green fluorescent protein-tagged-gephyrin and myc-tagged GlyR alpha1 subunits. In these experiments, myc-tagged GlyR alpha1 contained, or did not contain, the gephyrin-binding sequence (betagb) of the GlyR beta subunit. We report here that GlyR-gephyrin interaction localizes gephyrin to GlyR-containing organelles. Videomicroscopy and nocodazole treatment indicate that the movements of these vesicles are microtubule dependent. Expressing GlyR alpha1 with a thrombin cleavage site between the myc-tag and the N terminal of the GlyR alpha1 subunit (Rosenberg et al., 2001) allowed monitoring of newly inserted receptors in the cell surface. Using temperature changes to block GlyR in, and then release it from, the trans-Golgi network, we show that gephyrin accelerates the accumulation of GlyR at the cell surface. Therefore, our data strongly suggest that some GlyR clusters are associated with gephyrin on their way to the cell surface and that this association increases the accumulation of GlyR at the plasma membrane. PMID- 14762131 TI - Differential responses in human striatum and prefrontal cortex to changes in object and rule relevance. AB - Event-related functional magnetic resonance imaging was used to measure blood oxygenation level-dependent responses in 16 young healthy human volunteers during performance of an attentional switching task. The task allowed the separate investigation of lower-order switching between concrete objects and higher-order switching between abstract task rules. Significant signal change in the ventral striatum was demonstrated on trials when subjects switched between objects but not when subjects switched between abstract task rules. In contrast, signal change in the lateral prefrontal cortex (PFC) was observed during all switch trials. The switch-related responses were not contaminated by task difficulty, because the greatest signal change was observed during the relatively easy switch trials, which required both lower-order and higher-order switching at the same time. The present data suggest that mechanisms of inhibitory response control in frontostriatal systems are organized according to distinct levels of abstraction. Specifically, the response selection computation carried by the ventral striatum, which projects to the orbitofrontal cortex and the medial PFC, is restricted to the transformation of concrete stimulus exemplar information into motor responses, whereas the adaptive function of the lateral PFC extends to the transformation of abstract task-rule representations into action. PMID- 14762133 TI - Regulation of dopamine D1 receptor function by physical interaction with the NMDA receptors. AB - Functional interactions between dopamine D1-like receptors and NMDA subtype glutamate receptors have been implicated in the maintenance of normal brain activity and neurological dysfunction. Although modulation of NMDA receptor functions by D1 receptor activation has been the subject of extensive investigation, little is known as to how the activation of NMDA receptors alters D1 function. Here we report that NMDA receptors regulate D1 receptor function via a direct protein-protein interaction mediated by the carboxyl tail regions of both receptors. In both cotransfected cells and cultured hippocampal neurons the activation of NMDA receptors increases the number of D1 receptors on the plasma membrane surface and enhances D1 receptor-mediated cAMP accumulation via a SNARE dependent mechanism. Furthermore, overexpression of mini-genes encoding either NR1 or D1 carboxyl tail fragments disrupts the D1-NR1 direct protein-protein interaction and abolishes NMDA-induced changes in both D1 cell surface expression and D1-mediated cAMP accumulation. Our results demonstrate that the D1-NR1 physical interaction enables NMDA receptors to increase plasma membrane insertion of D1 receptors and provides a novel mechanism by which the activation of NMDA receptors upregulates D1 receptor function. Understanding the molecular mechanisms by which D1 and NMDA receptors functionally interact may provide insight toward elucidating the molecular neurobiological mechanisms involved in many neuropsychiatric illnesses, such as schizophrenia. PMID- 14762134 TI - Dynamics of precise spike timing in primary auditory cortex. AB - Although single units in primary auditory cortex (A1) exhibit accurate timing in their phasic response to the onset of sound (precision of a few milliseconds), paradoxically, they are unable to sustain synchronized responses to repeated stimuli at rates much beyond 20 Hz. To explore the relationship between these two aspects of cortical response, we designed a broadband stimulus with a slowly modulated spectrotemporal envelope riding on top of a rapidly modulated waveform (or fine structure). Using this stimulus, we quantified the ability of cortical cells to encode independently and simultaneously the stimulus envelope and fine structure. Specifically, by reverse-correlating unit responses with these two stimulus dimensions, we measured the spectrotemporal response fields (STRFs) associated with the processing of the envelope, the fine structure, and the complete stimulus. A1 cells respond well to the slow spectrotemporal envelopes and produce a wide variety of STRFs. In over 70% of cases, A1 units also track the fine-structure modulations precisely, throughout the stimulus, and for frequencies up to several hundred Hertz. Such a dual response, however, is contingent on the cell being driven by both fast and slow modulations, in that the response to the slowly modulated envelope gates the expression of the fine structure. We also demonstrate that either a simplified model of synaptic depression and facilitation, and/or a cortical network of thalamic excitation and cortical inhibition can account for major trends in the observed findings. Finally, we discuss the potential functional significance and perceptual relevance of these coexistent, complementary dynamic response modes. PMID- 14762132 TI - The glutamate transporter GLT1a is expressed in excitatory axon terminals of mature hippocampal neurons. AB - GLT1 is the major glutamate transporter of the brain and has been thought to be expressed exclusively in astrocytes. Although excitatory axon terminals take up glutamate, the transporter responsible has not been identified. GLT1 is expressed in at least two forms varying in the C termini, GLT1a and GLT1b. GLT1 mRNA has been demonstrated in neurons, without associated protein. Recently, evidence has been presented, using specific C terminus-directed antibodies, that GLT1b protein is expressed in neurons in vivo. These data suggested that the GLT1 mRNA detected in neurons encodes GLT1b and also that GLT1b might be the elusive presynaptic transporter. To test these hypotheses, we used variant-specific probes directed to the 3'-untranslated regions for GLT1a and GLT1b to perform in situ hybridization in the hippocampus. Contrary to expectation, GLT1a mRNA was the more abundant form. To investigate further the expression of GLT1 in neurons in the hippocampus, antibodies raised against the C terminus of GLT1a and against the N terminus of GLT1, found to be specific by testing in GLT1 knock-out mice, were used for light microscopic and EM-ICC. GLT1a protein was detected in neurons, in 14-29% of axons in the hippocampus, depending on the region. Many of the labeled axons formed axo-spinous, asymmetric, and, thus, excitatory synapses. Labeling also occurred in some spines and dendrites. The antibody against the N terminus of GLT1 also produced labeling of neuronal processes. Thus, the originally cloned form of GLT1, GLT1a, is expressed as protein in neurons in the mature hippocampus and may contribute significantly to glutamate uptake into excitatory terminals. PMID- 14762135 TI - Functional magnetic resonance imaging examination of two modular architectures for switching multiple internal models. AB - An internal model is a neural mechanism that can mimic the input-output properties of a controlled object such as a tool. Recent research interests have moved on to how multiple internal models are learned and switched under a given context of behavior. Two representative computational models for task switching propose distinct neural mechanisms, thus predicting different brain activity patterns in the switching of internal models. In one model, called the mixture-of experts architecture, switching is commanded by a single executive called a "gating network," which is different from the internal models. In the other model, called the MOSAIC (MOdular Selection And Identification for Control), the internal models themselves play crucial roles in switching. Consequently, the mixture-of-experts model predicts that neural activities related to switching and internal models can be temporally and spatially segregated, whereas the MOSAIC model predicts that they are closely intermingled. Here, we directly examined the two predictions by analyzing functional magnetic resonance imaging activities during the switching of one common tool (an ordinary computer mouse) and two novel tools: a rotated mouse, the cursor of which appears in a rotated position, and a velocity mouse, the cursor velocity of which is proportional to the mouse position. The switching and internal model activities temporally and spatially overlapped each other in the cerebellum and in the parietal cortex, whereas the overlap was very small in the frontal cortex. These results suggest that switching mechanisms in the frontal cortex can be explained by the mixture-of experts architecture, whereas those in the cerebellum and the parietal cortex are explained by the MOSAIC model. PMID- 14762137 TI - Olfactory bulb glomeruli: external tufted cells intrinsically burst at theta frequency and are entrained by patterned olfactory input. AB - Glomeruli, the initial sites of synaptic processing in the olfactory system, contain at least three types of neurons collectively referred to as juxtaglomerular (JG) neurons. The role of JG neurons in odor processing is poorly understood. We investigated the morphology, spontaneous, and sensory-evoked activity of one class of JG neurons, external tufted (ET) cells, using whole-cell patch-clamp and extracellular recordings in rat olfactory bulb slices. ET cells have extensive dendrites that ramify within a single glomerulus or, rarely, in two adjacent glomeruli. All ET neurons exhibit spontaneous rhythmic bursts of action potentials (approximately 1-8 bursts/sec). Bursting is intrinsically generated; bursting persisted and became more regular in the presence of ionotropic glutamate and GABA receptor antagonists. Burst frequency is voltage dependent; frequency increased at membrane potentials depolarized relative to rest and decreased during membrane potential hyperpolarization. Spontaneous bursting persisted in blockers of calcium channels that eliminated low-threshold calcium spikes (LTS) in ET cells. ET cells have a persistent sodium current available at membrane potentials that generate spontaneous bursting. Internal perfusion with a fast sodium channel blocker eliminated spontaneous bursting but did not block the LTS. These results suggest that persistent sodium channels are essential for spontaneous burst generation in ET cells. ET cell bursts were entrained to ON stimuli delivered over the range of theta frequencies. Thus, ET cells appear to be tuned to the frequency of sniffing. PMID- 14762138 TI - Macaque ventral premotor cortex exerts powerful facilitation of motor cortex outputs to upper limb motoneurons. AB - The ventral premotor area (F5) is part of the cortical circuit controlling visuomotor grasp. F5 could influence hand motor function through at least two pathways: corticospinal projections and corticocortical projections to primary motor cortex (M1). We found that stimulation of macaque F5, which by itself evoked little or no detectable corticospinal output, could produce a robust modulation of motor outputs from M1. Arrays of fine microwires were implanted in F5 and M1. During terminal experiments under chloralose anesthesia, single stimuli delivered to M1 electrodes evoked direct (D) and indirect (I1,I2, and I3) corticospinal volleys. In contrast, single F5 shocks were ineffective; double shocks (3 msec separation) evoked small I waves but no D wave. However, when the test (T) M1 shock was conditioned (C) by single or double F5 shocks, there was strong facilitation of I2 and I3 waves from M1, with C-T intervals of <1 msec. Intracellular recordings from 79 arm and hand motoneurons (MNs) revealed no postsynaptic effects from single F5 shocks. In contrast, these stimuli produced a robust facilitation of I2 and I3 EPSPs evoked from M1 (60% of MNs); this was particularly marked in hand muscle MNs (92%). Muscimol injection in M1 reduced I waves from F5 and abolished the F5-induced facilitation of late I waves from M1, and of EPSPs associated with them. Thus, some motor effects evoked from F5 may be mediated by corticocortical inputs to M1 impinging on interneurons generating late corticospinal I waves. Similar mechanisms may allow F5 to modulate grasp related outputs from M1. PMID- 14762136 TI - Unique neural circuitry for neonatal olfactory learning. AB - Imprinting ensures that the infant forms the caregiver attachment necessary for altricial species survival. In our mammalian model of imprinting, neonatal rats rapidly learn the odor-based maternal attachment. This rapid learning requires reward-evoked locus ceruleus (LC) release of copious amounts of norepinephrine (NE) into the olfactory bulb. This imprinting ends at postnatal day 10 (P10) and is associated with a dramatic reduction in reward-evoked LC NE release. Here we assess whether the functional emergence of LC alpha2 inhibitory autoreceptors and the downregulation of LC alpha1 excitatory autoreceptors underlie the dramatic reduction in NE release associated with termination of the sensitive period. Postsensitive period pups (P12) were implanted with either LC or olfactory bulb cannulas, classically conditioned with intracranial drug infusions (P14), and tested for an odor preference (P15). During conditioning, a novel odor was paired with either olfactory bulb infusion of abeta-receptor agonist (isoproterenol) to assess the target effects of NE or direct LC cholinergic stimulation combined with alpha2 antagonists and alpha1 agonists in a mixture to reinstate neonatal levels of LC autoreceptor activity to assess the source of NE. Pups learned an odor preference when the odor was paired with either olfactory bulb isoproterenol infusion or reinstatement of neonatal LC receptor activity. These results suggest that LC autoreceptor functional changes rather than olfactory bulb changes underlie sensitive period termination. PMID- 14762139 TI - A novel cAMP-dependent pathway activates neuronal integrin function in retinal neurons. AB - Retinal neurons lose the ability to attach to and extend neurites on substrata of laminin-1 (LN-1) during late embryogenesis, in a time frame that corresponds to target innervation. Although this developmental loss correlates with a modest downregulation of integrin expression, we have shown previously that these neurons use the same laminin-binding integrins for outgrowth on other laminin isoforms to which responsivity has not been lost (Ivins et al., 1998), suggesting that integrin functional states may be a critical point of regulation. Consistent with this view, expression of an activated mutant of R-ras, an activator of integrin function, restores integrin-dependent outgrowth of late embryonic retinal neurons on LN-1 (Ivins et al., 2000). Because cyclic nucleotides have been implicated in the regulation of integrin function in non-neuronal cells, as well as in the regulation of growth cone responses to various axon growth inhibitors, we asked whether raising cAMP levels in late embryonic retinal neurons could activate neuronal integrin function and restore neurite outgrowth on LN-1. We find that, similar to R-ras expression, raising cAMP levels in these neurons promotes alpha6beta1 integrin-dependent neurite outgrowth. Surprisingly, these effects of cAMP are independent of protein kinase A and the EPAC (exchange protein directly activated by cAMP)/Rap pathway and suggest the existence of a novel cAMP-dependent mechanism. PMID- 14762140 TI - Dopamine and glutamate control area-restricted search behavior in Caenorhabditis elegans. AB - Area-restricted search (ARS) is a foraging strategy used by many animals to locate resources. The behavior is characterized by a time-dependent reduction in turning frequency after the last resource encounter. This maximizes the time spent in areas in which resources are abundant and extends the search to a larger area when resources become scarce. We demonstrate that dopaminergic and glutamatergic signaling contribute to the neural circuit controlling ARS in the nematode Caenorhabditis elegans. Ablation of dopaminergic neurons eliminated ARS behavior, as did application of the dopamine receptor antagonist raclopride. Furthermore, ARS was affected by mutations in the glutamate receptor subunits GLR 1 and GLR-2 and the EAT-4 glutamate vesicular transporter. Interestingly, preincubation on dopamine restored the behavior in worms with defective dopaminergic signaling, but not in glr-1, glr-2, or eat-4 mutants. This suggests that dopaminergic and glutamatergic signaling function in the same pathway to regulate turn frequency. Both GLR-1 and GLR-2 are expressed in the locomotory control circuit that modulates the direction of locomotion in response to sensory stimuli and the duration of forward movement during foraging. We propose a mechanism for ARS in C. elegans in which dopamine, released in response to food, modulates glutamatergic signaling in the locomotory control circuit, thus resulting in an increased turn frequency. PMID- 14762141 TI - Ca2+ syntillas, miniature Ca2+ release events in terminals of hypothalamic neurons, are increased in frequency by depolarization in the absence of Ca2+ influx. AB - Localized, brief Ca2+ transients (Ca2+ syntillas) caused by release from intracellular stores were found in isolated nerve terminals from magnocellular hypothalamic neurons and examined quantitatively using a signal mass approach to Ca2+ imaging. Ca2+ syntillas (scintilla, L., spark, from a synaptic structure, a nerve terminal) are caused by release of approximately 250,000 Ca ions on average by a Ca2+ flux lasting on the order of tens of milliseconds and occur spontaneously at a membrane potential of -80 mV. Syntillas are unaffected by removal of extracellular Ca2+, are mediated by ryanodine receptors (RyRs) and are increased in frequency, in the absence of extracellular Ca2+, by physiological levels of depolarization. This represents the first direct demonstration of mobilization of Ca2+ from intracellular stores in neurons by depolarization without Ca2+ influx. The regulation of syntillas by depolarization provides a new link between neuronal activity and cytosolic [Ca2+] in nerve terminals. PMID- 14762142 TI - KCNQ2 is a nodal K+ channel. AB - Mutations in the gene encoding the K+ channel KCNQ2 cause neonatal epilepsy and myokymia, indicating that KCNQ2 regulates the excitability of CNS neurons and motor axons, respectively. We show here that KCNQ2 channels are functional components of axon initial segments and nodes of Ranvier, colocalizing with ankyrin-G and voltage-dependent Na+ channels throughout the CNS and PNS. Retigabine, which opens KCNQ channels, diminishes axonal excitability. Linopirdine, which blocks KCNQ channels, prolongs the repolarization of the action potential in neonatal nerves. The clustering of KCNQ2 at nodes and initial segments lags that of ankyrin-G during development, and both ankyrin-G and KCNQ2 can be coimmunoprecipitated in the brain. KCNQ3 is also a component of some initial segments and nodes in the brain. The diminished activity of mutant KCNQ2 channels accounts for neonatal epilepsy and myokymia; the cellular locus of these effects may be axonal initial segments and nodes. PMID- 14762143 TI - Efficacy of rehabilitative experience declines with time after focal ischemic brain injury. AB - To maximize the effectiveness of rehabilitative therapies after stroke, it is critical to determine when the brain is most responsive (i.e., plastic) to sensorimotor experience after injury and to focus such efforts within this period. Here, we compared the efficacy of 5 weeks of enriched rehabilitation (ER) initiated at 5 d (ER5), ER14, or ER30 after focal ischemia, as judged by functional outcome and neuromorphological change. ER5 provided marked improvement in skilled forelimb reaching ability and ladder-rung- and narrow-beam-walking tasks and attenuated the stroke-induced reliance on the unaffected forepaw for postural support. ER14 provided improvement to a somewhat lesser extent, whereas recovery was diminished after ER30 such that motor function did not differ from ischemic animals exposed to social housing. To examine potential neural substrates of the improved function, we examined dendritic morphology in the undamaged motor cortex because our previous work (Biernaskie and Corbett, 2001) suggested that recovery was associated with enhanced dendritic growth in this region. ER5 increased the number of branches and complexity of layer V neurons compared with both social housing and control animals. Dendritic arbor after ER14 (although increased) and ER30 did not differ from those exposed to social housing. These data suggest that the poststroke brain displays heightened sensitivity to rehabilitative experience early after the stroke but declines with time. These findings have important implications for rehabilitation of stroke patients, many of whom experience considerable delays before therapy is initiated. PMID- 14762145 TI - Recasting the smooth pursuit eye movement system. AB - Primates use a combination of smooth pursuit and saccadic eye movements to stabilize the retinal image of selected objects within the high-acuity region near the fovea. Pursuit has traditionally been viewed as a relatively automatic behavior, driven by visual motion signals and mediated by pathways that connect visual areas in the cerebral cortex to motor regions in the cerebellum. However, recent findings indicate that this view needs to be reconsidered. Rather than being controlled primarily by areas in extrastriate cortex specialized for processing visual motion, pursuit involves an extended network of cortical areas, and, of these, the pursuit-related region in the frontal eye fields appears to exert the most direct influence. The traditional pathways through the cerebellum are important, but there are also newly identified routes involving structures previously associated with the control of saccades, including the basal ganglia, the superior colliculus, and nuclei in the brain stem reticular formation. These recent findings suggest that the pursuit system has a functional architecture very similar to that of the saccadic system. This viewpoint provides a new perspective on the processing steps that occur as descending control signals interact with circuits in the brain stem and cerebellum responsible for gating and executing voluntary eye movements. Although the traditional view describes pursuit and saccades as two distinct neural systems, it may be more accurate to consider the two movements as different outcomes from a shared cascade of sensory motor functions. PMID- 14762144 TI - Pax6 and engrailed 1 regulate two distinct aspects of renshaw cell development. AB - Many of the interneuron cell types present in the adult spinal cord contribute to the circuits that control locomotion and posture. Little is known, however, about the embryonic origin of these cell types or the molecular mechanisms that control their differentiation. Here we provide evidence that V1 interneurons (INs), an embryonic class of interneurons that transiently express the En1 transcription factor, differentiate as local circuit inhibitory interneurons and form synapses with motor neurons. Furthermore, we show that a subset of V1 INs differentiates as Renshaw cells, the interneuronal cell type that mediates recurrent inhibition of motor neurons. We analyze the role that two V1 IN-related transcription factor genes play in Renshaw cell development. Pax6 (paired box gene 6) is necessary for an early step in Renshaw cell development, whereas Engrailed 1 (En1), which is genetically downstream of Pax6, regulates the formation of inhibitory synapses between Renshaw cells and motor neurons. Together, these results show that Pax6 and En1 have essential roles in establishing the recurrent inhibitory circuit between motor neurons and Renshaw cells. PMID- 14762146 TI - Short-term adaptation of auditory receptive fields to dynamic stimuli. AB - Short-term adaptation and recovery from adaptation have a strong impact on the processing of dynamic stimuli. Adaptive effects on neuronal activity have been studied most commonly for changes in first-order statistics of stimuli such as stepwise increments or decrements in stimulus amplitude. However, changes in higher moment statistics, such as the variance of the amplitude distribution in visual stimuli, also can invoke pronounced adaptation behavior. We demonstrate here that neurons in the inferior colliculus (ICC) of the cat show adaptation to dynamic auditory stimuli that differ in the variance of their modulation depth distribution. In addition, it is shown that neurons show adaptation to other higher moment statistics (e.g., kurtosis) of the modulation envelope. The time course of adaptation is specific for the altered stimulus property and the direction of parameter change. The use of dynamic stimuli allows an estimate of the effects of the adaptation on the temporal response properties of the neurons. We demonstrate that temporal receptive fields of neurons undergo change during the course of adaptation. We show that adaptation to variance in the ICC has many similarities to that in the retina and suggest that adaptation to variance is a general property of sensory systems that allows them to effectively deal with a nonstationary environment. PMID- 14762147 TI - Nitric oxide and histamine induce neuronal excitability by blocking background currents in neuron MCC of Aplysia. AB - Nitric oxide (NO) and histamine are important neurotransmitters and neuromodulators. We investigated their ability to modulate the membrane ionic currents and excitability of the metacerebral cell (MCC) of Aplysia using voltage clamp techniques. MCC is a serotonergic modulator of the feeding neural circuit. It receives powerful long-lasting excitatory synaptic input mediated by NO and histamine. NO donors reduced a background outward current at and above the resting potential, associated with decreased membrane conductance. This produced a substantial steady-state inward current that was relatively insensitive to cesium or cobalt. The NO response appears to be due to the reduction of a background potassium current and a small increase in persistent inward sodium current. Treatment with 8-bromoguanosine-3'5'-cyclic monophosphate mimics this response, suggesting it is mediated primarily by the NO-guanylyl cyclase-cGMP pathway. In some MCCs, NO blocked an additional potassium current that resulted in current reversal near the potassium equilibrium potential in current-voltage plots. Histamine also reduced a background outward current at and above the resting potential. However, treatment with cobalt, which blocks calcium and calcium-dependent currents, blocked the histamine response, suggesting that histamine decreases calcium activated potassium currents. Although nifedipine (L type calcium channel blocker) and tetraethylammonium reduced some calcium and calcium-dependent potassium currents, they had only a slight effect on the NO and histamine responses. Both NO and histamine decreased steady-state membrane currents, and thereby depolarized MCC and increased its excitability, but different ionic currents and second messenger pathways are involved, allowing complex state and time dependent modulation of MCC's activity. PMID- 14762148 TI - Dynamics of population rate codes in ensembles of neocortical neurons. AB - Information processing in neocortex can be very fast, indicating that neuronal ensembles faithfully transmit rapidly changing signals to each other. Apart from signal-to-noise issues, population codes are fundamentally constrained by the neuronal dynamics. In particular, the biophysical properties of individual neurons and collective phenomena may substantially limit the speed at which a graded signal can be represented by the activity of an ensemble. These implications of the neuronal dynamics are rarely studied experimentally. Here, we combine theoretical analysis and whole cell recordings to show that encoding signals in the variance of uncorrelated synaptic inputs to a neocortical ensemble enables faithful transmission of graded signals with high temporal resolution. In contrast, the encoding of signals in the mean current is subject to low-pass filtering. PMID- 14762149 TI - Role of persistent sodium and calcium currents in motoneuron firing and spasticity in chronic spinal rats. AB - After chronic spinal injury, motoneurons spontaneously develop two persistent inward currents (PICs): a TTX-sensitive persistent sodium current (sodium PIC) and a nimodipine-sensitive persistent calcium current (calcium PIC). In the present paper, we examined how these PICs contributed to motoneuron firing. Adult rats were spinalized at the S(2) sacral level, and after 2 months intracellular recordings were made from sacrocaudal motoneurons in vitro. The PICs and repetitive firing were measured with slow triangular voltage and current ramps, respectively. The sodium PIC was examined after blocking the calcium PIC with nimodipine (20 microM; n = 12). It was always activated subthreshold, and during current ramps in nimodipine, it produced a sodium plateau that assisted in initiating and maintaining firing (self-sustained firing). The sodium PIC oscillated off and on during firing and helped initiate each spike, and near threshold this caused abnormally slow firing (2.82 +/- 1.21 Hz). A low dose of TTX (0.5 microM) blocked the sodium PIC, sodium plateau, and very slow firing prior to affecting the spike itself. The calcium PIC was estimated as the current blocked by nimodipine or current remaining in TTX (2 microM; n = 13). In 59% of motoneurons, the calcium PIC was activated subthreshold to firing and produced a plateau that assisted in initiating and sustaining firing because nimodipine significantly increased the firing threshold current and decreased the self sustained firing. In the remaining motoneurons (41%), the calcium PIC was activated suprathreshold to firing and during current ramps did not initially affect firing but eventually was activated and caused an acceleration in firing followed by self-sustained firing, which were blocked by nimodipine. The frequency-current (F-I) slope was 3.0 +/- 1.0 Hz/nA before the calcium PIC activation (primary range), 6.3 +/- 3.6 Hz/nA during the calcium PIC onset (secondary range; acceleration), and 2.1 +/- 1.3 Hz/nA with the calcium PIC steadily activated (tertiary range). Nimodipine eliminated the secondary and tertiary ranges, leaving a linear F-I slope of 3.7 +/- 1.0 Hz/nA. A single low threshold shock to the dorsal root evoked a many-second-long discharge, the counterpart of a muscle spasm in the awake chronic spinal rat. This long-lasting reflex was caused by the motoneuron PICs because when the activation of the voltage-dependent PICs was prevented by hyperpolarization, the same dorsal root stimulation only produced a brief excitatory postsynaptic potential (<1 s). Both the calcium and sodium PIC were involved because nimodipine only partly reduced the reflex and there remained very slow firing mediated by the sodium PIC. PMID- 14762150 TI - Neuronal activity in the primate substantia nigra pars reticulata during the performance of simple and memory-guided elbow movements. AB - The basal ganglia participate in motor functions and are implicated in the pathophysiology of movement disorders. It has been shown in primates that the activity of many neurons in one of the basal ganglia output nuclei, the internal segment of the globus pallidus, changes with active or passive movements. The involvement of the second major output nucleus, the substantia nigra pars reticulata (SNr), in movement is less well established. In this study, the electrophysiologic activity of SNr neurons was studied in two awake Rhesus monkeys while the animals were examined and while they performed elbow movements in two different motor tasks (n = 261 cells). Responses to examination were uncommon and subtle. Twenty-one percent of neurons responded to the target step in a step tracking task, mostly with anticipatory responses, although some cells showed directional, movement-related activity. In a delayed-response task, 17% of cells showed anticipatory activity to an instruction cue preceding the target jump, 11% responded directly to the cue, and 11% showed long-lasting postcue activity. Movement-related responses were seen in 21% in this task. Reward responses occurred in 10% of neurons. Responses to more than one event were common. The results demonstrate that few neurons in the SNr respond directly to passive or active movements, but a large proportion shows responses that may be related to memory, attention, or movement preparation. While internal pallidal segment neurons may be preferentially concerned with controlling elemental movement parameters, neurons in the SNr may be more involved in higher motor functions or nonmotor aspects of behavior. PMID- 14762151 TI - Activation of ionotropic glutamate receptors on peripheral axons of primary motoneurons mediates transmitter release at the zebrafish NMJ. AB - The development and function of the vertebrate neuromuscular junction (NMJ) is continually being redefined. Previous studies have indicated that glutamate may play a role in the development or function of the NMJ by associating with presynaptic receptors. We have used larval zebrafish (Danio rerio) to investigate the presence of presynaptic ionotropic glutamate receptors (iGluRs) at the NMJ in vivo. In whole-mount zebrafish larvae, antibody staining directed to NR2A subunits colocalized with specific staining of motoneuron axon tracts. Whole cell voltage-clamp recordings of miniature endplate currents (mEPCs) from axial white muscle were performed during application of iGluR agonists and antagonists. Local perfusion of the NMJ with iGluR agonists resulted in significant increases in the frequency of spontaneous acetylcholine (ACh) release. These increases were blocked by the N-methyl-d-aspartate (NMDA) receptor antagonist d-(-)-2-amino-5 phosphonopentanoic acid (50 microM) and by the non-NMDA receptor antagonist 6 cyano-7-nitroquinoxalene-2,3-dione (50 microM). Further pharmacological investigation revealed no effect of the kainate receptor-specific antagonist (2S,4R)-4-methylglutamate (10 microM) on kainate-induced rises in the frequency of spontaneous ACh release. However, these were blocked with the AMPA receptor specific antagonist 1-(4-aminophenyl)-4-methyl-7,8-methylenedioxy-5H-2,3 benzodiazepine (50 microM). Application of glutamate (1 mM) in the presence of the glutamate uptake inhibitor d-threo-beta-benzyloxyaspartate(200 microM) resulted in a significant increase in the frequency of mEPCs. These results suggest the presence of AMPA and NMDA receptors in association with motoneuron axons of larval zebrafish. PMID- 14762152 TI - Nicotinic acetylcholine receptors in mouse and rat optic nerves. AB - Receptor-mediated calcium signaling in axons of mouse and rat optic nerves was examined by selectively staining the axonal population with a calcium indicator. Nicotine (1-50 microM) induced an axonal calcium elevation that was eliminated when calcium was removed from the bath, suggesting that nicotine induces calcium influx into axons. The nicotine response was blocked by d-tubocurarine and mecamylamine but not alpha-bungarotoxin, indicating the presence of calcium permeable, non-alpha7 nicotinic acetylcholine receptor (nAChR) subtype. Agonist efficacy order for eliciting the axonal nAChR calcium response was cytisine approximately nicotine >> acetylcholine. The nicotine-mediated calcium response was attenuated during the process of normal myelination, decreasing by approximately 10-fold from P1 (premyelinated) to P30 (myelinated). Nicotine also caused a rapid reduction in the compound action potential in neonatal optic nerves, consistent with a shunting of the membrane after opening of the nonspecific cationic nicotinic channels. Voltagegated calcium channels contributed little to the axonal calcium elevation during nAChR activation. During repetitive stimulations, the compound action potential in neonatal mouse optic nerves underwent a gradual reduction in amplitude that could be partially prevented by d-tubocurarine, suggesting an activity-dependent release of acetylcholine that activates axonal AChRs. We conclude that mammalian optic nerve axons express nAChRs and suggest that these receptors are activated in an activity-dependent fashion during optic nerve development to modulate axon excitability and biology. PMID- 14762153 TI - Silent synapses in the immature visual cortex: layer-specific developmental regulation. AB - Central glutamatergic synapses are thought to initially form as immature, so called silent synapses showing exclusively N-methyl-d-aspartate receptor-mediated synaptic transmission. Postsynaptic insertion of alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionic acid receptors during further development leads to a conversion into functional, mature synapses. Here, we tested the hypothesis that, according to the "inside first-outside last" pattern of neocortical layer formation and synaptogenesis, pyramidal cells in the superficial layers might show a higher fraction of silent synapses compared with pyramidal cells in the deep layers. We performed an electrophysiological analysis of glutamatergic synapses in acute rat visual cortex slices during postnatal development. In layer VI pyramidal neurons the incidence of silent synapses was high during the first postnatal week and strongly declined during further development. Surprisingly, in superficial cortical plate pyramidal neurons (immature layers II/III), the fraction of silent synapses was initially very low and increased up to the second postnatal week. Thereafter, a similar decline as found in layer VI pyramidal neurons was observed. Thus the developmental regulation of silent synapses was clearly different in pyramidal neurons from different neocortical layers. The almost complete absence of silent synapses at early stages in layer II/III pyramidal neurons indicates that an initially formed subset of synapses is constitutively functional. This might be important to enable spontaneous activity and latter activity-dependent maturation of synapses. PMID- 14762154 TI - Two types of C nociceptors in human skin and their behavior in areas of capsaicin induced secondary hyperalgesia. AB - Peripheral nociceptor sensitization is accepted as an important mechanism of cutaneous primary hyperalgesia, but secondary hyperalgesia has been attributed to central mechanisms since evidence for sensitization of primary afferents has been lacking. In this study, microneurography was used to test for changes in sensitivity of C nociceptors in the area of secondary hyperalgesia caused by intradermal injection of capsaicin in humans. Multiple C units were recruited by electrical stimulation of the skin at 0.25 Hz and were identified as discrete series of dots in raster plots of spike latencies. Nociceptors slowed progressively during repetitive stimulation at 2 Hz for 3 min. According to their response to mechanical stimulation, nociceptors could be classified as either mechano-sensitive (CM) or mechano-insensitive (CM(i)). These two nociceptor subtypes had different axonal properties: CM(i) units slowed by 2% or more when stimulated at 0.25 Hz after a 3-min pause, whereas CM units slowed by <1%. This stimulation protocol was used before capsaicin injection to identify nociceptor subtype without repeated probing, thus avoiding possible mechanical sensitization. Capsaicin, injected 10-50 mm away from the site of electrical stimulation, had no effect on any of 29 CM units, but induced bursts of activity in 11 of 15 CM(i) units, after delays ranging from 0.5 to 18 min. The capsaicin injections also sensitized a majority of the CM(i) units, so that 11 of 17 developed immediate or delayed responsiveness to mechanical stimuli. This sensitization may contribute a peripheral C fiber component to secondary hyperalgesia. PMID- 14762155 TI - Monkey hand postural synergies during reach-to-grasp in the absence of vision of the hand and object. AB - Understanding how the CNS controls reach-to-grasp will require behavioral and neurophysiological studies of reach-to-grasp in the monkey, including the evaluation of whole-hand grasp with explicit force requirements. In this study, monkeys performed a reach-to-grasp task in which the size, shape, and orientation of the objects were varied. The monkeys were required to grasp each object at five force levels based on visual feedback. Seventeen positions on the wrist and hand were monitored to quantify kinematics. Hand shaping began with initiation of reach and continued throughout the reach, matching object properties even without vision of the hand or object. Grasp aperture scaled to object size. Singular value decomposition analysis of the marker positions identified two dominant hand postures. The first eigenvector or "eigenposture" consisted of an open hand configuration midway between flexion and extension that explained >93% of the variance. The second eigenposture consisted of hyperextension of all joints that accounted for another 4-5% of the variance. The two eigenpostures were similar across force levels and between monkeys. Reach kinematics consisted of a U-shaped hand path with a bell-shaped velocity profile. Trajectory and speed were independent of grasp force and object properties. In summary, hand shaping during the reach occurred without vision of the hand or object, and hand kinematics were not dependent on grasp force. Furthermore, the reach was independent of grasp force and object properties. These observations imply that the kinematics of reach-to-grasp and grasp force are controlled independently. Similar to humans, monkeys may use a simplifying strategy to reduce the degrees of freedom of the hand during reach-to-grasp. PMID- 14762156 TI - Mechanisms for the modulation of native glycine receptor channels by ethanol. AB - Previously, we showed that ethanol increases synaptic glycine currents, an effect that depends on ethanol concentration and developmental age of the preparation. Glycine receptor (GlyR) subunits undergo a shift from alpha2/beta to alpha1/beta from neonate to juvenile ages, with synaptic glycine currents from neonate hypoglossal motoneurons (HMs) being less sensitive to ethanol than those from juvenile HMs. Here we investigate whether these dose and developmental effects are also present in excised membrane patches containing GlyRs and if ethanol changes response kinetics. We excised outside-out patches from rat HM somata and applied glycine using either a picospritzer or piezo stack translator. Ethanol (100 mM) increased the response to glycine (200 microM) of patches from neonate and juvenile HMs. However, 30 mM ethanol increased the response from only juvenile HM patches. Using a lower concentration of glycine (30 microM) to observe single channel openings, we found that 100 mM ethanol increased the number of GlyRs that open in response to glycine and decreased first latency to channel opening. To investigate GlyR kinetic properties, we rapidly applied 1 mM glycine for 1 ms and found that glycine currents were increased by ethanol (100 mM) at both ages. For patches from juvenile HMs, ethanol consistently decreased response rise-time and increased response decay time. Using kinetic modeling, we determined that ethanol's potentiation of the glycine response arises from an increase in the glycine association (k(on)) and a decrease in the dissociation (k(off)) rate constants, resulting in increased glycine affinity of the GlyR. PMID- 14762157 TI - Influence of contrast on orientation and temporal frequency tuning in ferret primary visual cortex. AB - Neurons in primary visual cortex are highly sensitive to the contrast, orientation, and temporal frequency of a visual stimulus. These three stimulus properties can be varied independently of one another, raising the question of how they interact to influence neuronal responses. We recorded from individual neurons in ferret primary visual cortex to determine the influence of stimulus contrast on orientation tuning, temporal-frequency tuning, and latency to visual response. Results show that orientation-tuning bandwidth is not affected by contrast level. Thus neurons in ferret visual cortex display contrast-invariant orientation tuning. Stimulus contrast does, however, influence the structure of orientation-tuning curves as measures of circular variance vary inversely with contrast for both simple and complex cells. This change in circular variance depends, in part, on a contrast-dependent change in the ratio of null to preferred orientation responses. Stimulus contrast also has an influence on the temporal-frequency tuning of cortical neurons. Both simple and complex cells display a contrast-dependent rightward shift in their temporal frequency-tuning curves that results in an increase in the highest temporal frequency needed to produce a half-maximum response (TF(50)). Results show that the degree of the contrast-dependent increase in TF(50) is similar for cortical neurons and neurons in the lateral geniculate nucleus (LGN) and indicate that subcortical mechanisms likely play a major role in establishing the degree of effect displayed by downstream neurons. Finally, results show that LGN and cortical neurons experience a contrast-dependent phase advance in their visual response. This phase advance is most pronounced for cortical neurons indicating a role for both subcortical and cortical mechanisms. PMID- 14762158 TI - Ocular tracking of moving targets: effects of perturbing the background. AB - Primates are able to track a moving target with their eyes, even when the target is seen against a stationary textured background. In this situation, the tracking eye movement induces motion of the background images on the retina (reafference) that competes with the motion of the target's retinal image, potentially disrupting the tracking of the target. Previous work on humans reported that brief perturbations of the background in the opposite direction to pursuit were much less disruptive than perturbations in the same direction as pursuit. Furthermore, if the background moved together with the pursuit target--so as to effectively eliminate the reafference--then the effects of a subsequent background perturbation showed less dependence on direction. This suggested that the direction selectivity to background perturbations during pursuit against a stationary background was due, at least in part, to the prior motion of the background secondary to the pursuit. We now report similar findings in monkeys, and in addition, have investigated the effect of moving the background while the animal was fixating a stationary target. In this situation, the ocular tracking responses to subsequent brief perturbations of the moving background were weaker when the perturbations were in the same direction as the prior background motion than when in the opposite direction. This suggests that the selective insensitivity to the reafferent visual input associated with pursuit across a stationary background is, at least in part, independent of pursuit per se and attributable to a progressive reduction in the sensitivity to sustained background motion. PMID- 14762159 TI - Enhanced long-term potentiation during aging is masked by processes involving intracellular calcium stores. AB - The contribution of Ca(2+) release from intracellular Ca(2+) stores (ICS) for regulation of synaptic plasticity thresholds during aging was investigated in hippocampal slices of old (22-24 mo) and young adult (5-8 mo) male Fischer 344 rats. Inhibition of Ca(2+)-induced Ca(2+) release by thapsigargin, cyclopiazonic acid (CPA), or ryanodine during pattern stimulation near the threshold for synaptic modification (5 Hz, 900 pulses) selectively induced long-term potentiation (LTP) to CA1 Schaffer collateral synapses of old rats. Increased synaptic strength was specific to test pathways and blocked by AP-5. Intracellular recordings demonstrated that ICS plays a role in the augmentation of the afterhyperpolarization (AHP) in old rats. The decrease in the AHP by ICS inhibition was reversed by the L-channel agonist, Bay K8644. Under conditions of ICS inhibition and a Bay K8644-mediated enhancement of the AHP, pattern stimulation failed to induce LTP, consistent with the idea that the AHP amplitude shapes the threshold for LTP induction. Finally, ICS inhibition was associated with an increase in the N-methyl-d-aspartate (NMDA) receptor component of synaptic transmission in old animals. This increase in the synaptic response was blocked by the calcineurin inhibitor FK506. The results reveal an age-related increase in susceptibility to LTP-induction that is normally inhibited by ICS and suggest that the age-related shift in Ca(2+) regulation and Ca(2+)-dependent synaptic plasticity is coupled to changes in cell excitability and NMDA receptor function through ICS. PMID- 14762160 TI - Reorganization of reflex responses mediated by different afferent sensory fibers after spinal cord transection. AB - Adult rats were submitted to a complete spinal cord transection at T9 level to address peripheral and spinal reflex changes in the caudal lumbar segments. Compound muscle and nerve action potentials decreased in amplitude and increased their duration between 14 and 30 days but recovered to near to normal values thereafter. The H wave amplitude increased during follow-up, resulting in significantly higher H/M ratio in tibialis anterior (223%), gastrocnemius (160%), and plantar (304%) muscles with respect to preoperatory values (P < 0.01). Sixty minutes after spinal cord transection, component C1 (conveyed by Aalphabeta afferents) disappeared in the crossed but not in the ipsilateral withdrawal reflex. Components C2 (Adelta) and C3 (C afferents) were abolished on both. C1 and C3 reappeared for both reflexes in all injured animals, while C2 reappeared in a few cases. C1 ipsilateral component became highly facilitated (209% of presurgery values, P < 0.01), whereas C3 (82%) and C2 (24%) recovered partially. Crossed reflex component C1 attained in all animals similar to normal values (85%) but with longer duration. C3 increased with time although it remained significantly lower than the original (67%) whereas C2 reappeared in only 2/8 animals. In conclusion, spinal cord injury induces a transient disability of caudal spinal cord segments that progressively reverts along time. Ipsilateral reflex components mediated by thick Aalphabeta fibers (H reflex and C1) but not those mediated by thin fibers (C2 and C3) remained present after injury showing long-lasting facilitation whereas contralateral reflex components were abolished after injury and showed limited recovery. PMID- 14762161 TI - Contextual modulation of central thalamic delay-period activity: representation of visual and saccadic goals. AB - This study examines the influence of behavioral context on the activity of visuomotor neurons in primate central thalamus. Neurons that combine information about sensory stimuli and their behavioral relevance are thought to contribute to the decision mechanisms that link specific stimuli to specific responses. We reported in a previous study that neurons in central thalamus carry spatial information throughout the instructed delay period of a visually guided delayed saccade task. The goal of the current study was to determine whether the delay period activity of thalamic neurons is modulated by behavioral context. Single neurons were evaluated during performance of visually guided and memory-guided variants of a saccadic choice task in which a cue designated the response field stimulus as the target of a rewarded saccade or as an irrelevant distracter. The relative influence of the physical stimulus and context on delay-period activity suggested a minimum of 3 neural groups. Some neurons signaled the locations of visible stimuli regardless of behavioral relevance. Other neurons preferentially signaled the locations of current saccadic goals and did so even in the absence of the physical stimulus. A third group signaled only the locations of currently visible saccadic goals. For the latter 2 groups, activity was the product of both stimulus and context, suggesting that central thalamic neurons play a role in the context-dependent linkage of sensory signals and saccadic commands. More generally, these data suggest that the anatomical substrate of sensorimotor decision making may include the cortico-subcortical loops for which central thalamus serves as the penultimate synapse. PMID- 14762162 TI - Results of a PNAS author survey on an open access option for publication. PMID- 14762163 TI - Preferential oxidation of the second phosphatase domain of receptor-like PTP alpha revealed by an antibody against oxidized protein tyrosine phosphatases. AB - Protein tyrosine phosphatases (PTPs) constitute a large enzyme family with important biological functions. Inhibition of PTP activity through reversible oxidation of the active-site cysteine residue is emerging as a general, yet poorly characterized, regulatory mechanism. In this study, we describe a generic antibody-based method for detection of oxidation-inactivated PTPs. Previous observations of oxidation of receptor-like PTP (RPTP) alpha after treatment of cells with H(2)O(2) were confirmed. Platelet-derived growth factor (PDGF)-induced oxidation of endogenous SHP-2, sensitive to treatment with the phosphatidylinositol 3-kinase inhibitor LY294002, was demonstrated. Furthermore, oxidation of RPTPalpha was shown after UV-irradiation. Interestingly, the catalytically inactive second PTP domain of RPTPalpha demonstrated higher susceptibility to oxidation. The experiments thus demonstrate previously unrecognized intrinsic differences between PTP domains to susceptibility to oxidation and suggest mechanisms for regulation of RPTPs with tandem PTP domains. The antibody strategy for detection of reversible oxidation is likely to facilitate further studies on regulation of PTPs and might be applicable to analysis of redox regulation of other enzyme families with active-site cysteine residues. PMID- 14762164 TI - Inducible, reversible, and stable RNA interference in mammalian cells. AB - RNA interference is a powerful genetic approach for efficiently silencing target genes. The existing method of gene suppression by the constitutive expression of short hairpin RNAs (shRNAs) allows analysis of the consequences of stably silencing genes but limits the analysis of genes essential for cell survival, cell cycle regulation, and cell development. We have developed an inducible U6 promoter for synthesis of shRNAs in both human and murine cells. Cells containing stably integrated shRNA expression constructs demonstrate stringent dosage- and time-dependent kinetics of induction with undetectable background expression in the absence of the inducer ecdysone. Inducible suppression of human p53 in glioblastoma cells shows striking morphological changes and defects in cell cycle arrest caused by DNA damage, as expected. Remarkably, the inducibility is reversible after withdrawal of the inducer, as observed by reappearance of the protein and a restoration of the original cell phenotype. Inducible and reversible regulation of RNA interference has broad applications in the areas of mammalian genetics and molecular therapeutics. PMID- 14762165 TI - A capsid protein of nonenveloped Bluetongue virus exhibits membrane fusion activity. AB - The outer capsid layer of Bluetongue virus, a member of the nonenveloped Reoviridae family, is composed of two proteins, a receptor-binding protein, VP2, and a second protein, VP5, which shares structural features with class I fusion proteins of enveloped viruses. In the replication cycle of Bluetongue virus VP5 acts as a membrane permeabilization protein that mediates release of viral particles from endosomal compartments into the cytoplasm. Here, we show that VP5 can also act as a fusion protein and induce syncytium formation when it is fused to a transmembrane anchor and expressed on the cell surface. Fusion activity is strictly pH-dependent and is triggered by short exposure to low pH. No cell-cell fusion is observed at neutral pH. Deletion of the first 40 amino acids, which can fold into two amphipathic helices, abolishes fusion activity. Syncytium formation by VP5 is inhibited in the presence of VP2 when it is expressed in a membrane anchored form. The data indicate an interaction between the outer capsid protein VP2 and VP5 and show that VP5 undergoes pH-dependent conformational changes that render it capable of interacting with cellular membranes. More importantly, our data show that a membrane permeabilization protein of a nonenveloped virus can evolve into a fusion protein by the addition of an appropriate transmembrane anchor. The results strongly suggest that the mechanism of membrane permeabilization by VP5 and membrane fusion by viral fusion proteins require similar structural features and conformational changes. PMID- 14762166 TI - IL-15 enhances the in vivo antitumor activity of tumor-reactive CD8+ T cells. AB - IL-15 and IL-2 possess similar properties, including the ability to induce T cell proliferation. However, whereas IL-2 can promote apoptosis and limit CD8(+) memory T cell survival and proliferation, IL-15 helps maintain a memory CD8(+) T cell population and can inhibit apoptosis. We sought to determine whether IL-15 could enhance the in vivo function of tumor/self-reactive CD8(+) T cells by using a T cell receptor transgenic mouse (pmel-1) whose CD8(+) T cells recognize an epitope derived from the self/melanoma antigen gp100. By removing endogenous IL 15 by using tumor-bearing IL-15 knockout hosts or supplementing IL-15 by means of exogenous administration, as a component of culture media or as a transgene expressed by adoptively transferred T cells, we demonstrate that IL-15 can improve the in vivo antitumor activity of adoptively transferred CD8(+) T cells. These results provide several avenues for improving adoptive immunotherapy of cancer in patients. PMID- 14762168 TI - Crossmaps: visualization of overlapping relationships in collections of journal papers. AB - A crossmapping technique is introduced for visualizing multiple and overlapping relations among entity types in collections of journal articles. Groups of entities from two entity types are crossplotted to show correspondence of relations. For example, author collaboration groups are plotted on the x axis against groups of papers (research fronts) on the y axis. At the intersection of each pair of author group/research front pairs a circular symbol is plotted whose size is proportional to the number of times that authors in the group appear as authors in papers in the research front. Entity groups are found by agglomerative hierarchical clustering using conventional similarity measures. Crossmaps comprise a simple technique that is particularly suited to showing overlap in relations among entity groups. Particularly useful crossmaps are: research fronts against base reference clusters, research fronts against author collaboration groups, and research fronts against term co-occurrence clusters. When exploring the knowledge domain of a collection of journal papers, it is useful to have several crossmaps of different entity pairs, complemented by research front timelines and base reference cluster timelines. PMID- 14762167 TI - B cell development and immunoglobulin transcription in Oct-1-deficient mice. AB - The POU domain transcription factors Oct-1 and Oct-2 interact with the octamer element, a motif conserved within Ig promoters and enhancers, and mediate transcription from the Ig loci. Inactivation of Oct-2 by gene targeting results in normal B cell development and Ig transcription. To study the role of Oct-1 in these processes, the lymphoid compartment of RAG-1(-/-) animals was reconstituted with Oct-1-deficient fetal liver hematopoietic cells. Recipient mice develop B cells with levels of surface Ig expression comparable with wild type, although at slightly reduced numbers. These B cells transcribe Ig normally, respond to antigenic stimulation, undergo class switching, and use a normal repertoire of light chain variable segments. However, recipient mice show slight reductions in serum IgM and IgA. Thus, the Oct-1 protein is dispensable for B cell development and Ig transcription. PMID- 14762169 TI - Small potassium ion channel proteins encoded by chlorella viruses. AB - Kcv, a 94-aa protein encoded by Paramecium bursaria chlorella virus 1, is the smallest known protein to form a functional potassium ion channel and basically corresponds to the "pore module" of potassium channels. Both viral replication and channel activity are inhibited by the ion channel blockers barium and amantadine but not by cesium. Genes encoding Kcv-like proteins were isolated from 40 additional chlorella viruses. Differences in 16 of the 94 amino acids were detected, producing six Kcv-like proteins with amino acid substitutions occurring in most of the functional domains of the protein (N terminus, transmembrane 1, pore helix, selectivity filter, and transmembrane 2). The six proteins form functional potassium selective channels in Xenopus oocytes with different properties including altered current kinetics and inhibition by cesium. The amino acid changes together with the different properties observed in the six Kcv-like channels will be used to guide site-directed mutations, either singularly or in combination, to identify key amino acids that confer specific properties to Kcv. PMID- 14762170 TI - Estrogen receptors ER alpha and ER beta in proliferation in the rodent mammary gland. AB - Most evidence supports the view that ER alpha is responsible for estrogen (ovarian estradiol, E(2))-induced proliferation in the epithelial cells of the mammary gland, but despite this, proliferating epithelial cells do not express ER alpha. We have examined this apparent paradox by studying the role of ER alpha and ER beta in E(2)-induced proliferation in mammary glands (measured by BrdUrd incorporation into DNA) in mice with intact ER beta (WT mice) and those in which the ER beta gene has been inactivated (ER beta(-/-) mice). On treatment of ER beta(-/-) mice with E(2) or ovariectomized WT mice with E(2), tamoxifen, or a specific ER beta agonist (BAG), the number of BrdUrd-labeled cells in mammary glands increased from 3.4% in controls to 28-38% in the treated mice. This indicates that both ER alpha and ER beta can mediate E(2)-induced proliferation independently of each other. With specific antibodies, ER beta was found in both epithelial and stromal cells, whereas ER alpha was strictly epithelial. Within 4 h of a single dose of E(2), ER alpha was lost from the nuclei of epithelial cells. In WT mice, ER alpha reappeared by 24 h, but in ER beta(-/-) mice, return to the nucleus was delayed by 24 h. At 4 h after E(2), neither ER alpha nor progesterone receptor was detectable in BrdUrd-labeled nuclei but by 48 h after E(2), 29% of the BrdUrd-labeled cells expressed ER alpha, and 21-38% expressed progesterone receptor. During 3 weeks of continuous E(2) treatment, ER beta remained in the nucleus, but there was no detectable ER alpha. With tamoxifen treatment, ER alpha remained in the nucleus, but ER beta was lost. From these results, we conclude that ER alpha receives the proliferation signal from E(2), initiates DNA synthesis, and is then lost from cells. The subsequent steps in proliferation can proceed in the absence of either ER alpha or ER beta. ER beta facilitates the return of ER alpha to the nucleus and restores responsiveness to E(2). By down-regulating ER beta, tamoxifen may prolong refractoriness to E(2) in mammary epithelium. PMID- 14762171 TI - Cofilin peptide homologs interfere with immunological synapse formation and T cell activation. AB - The formation of supramolecular activation clusters within the immunological synapse, crucial for sustained signaling and T lymphocyte activation, requires costimulation-dependent reorganization of the actin cytoskeleton. Here we have identified the actin-remodeling protein cofilin as a key player in this process. Cell-permeable peptides that block costimulation-induced cofilin/F-actin interactions in untransformed human T lymphocytes impair receptor capping and immunological synapse formation at the interface between T cells and antigen presenting cells. As a consequence, T cell activation, as measured by cytokine production and proliferation, is inhibited. PMID- 14762172 TI - Cholesterol feeding of mice expressing cholesterol 7alpha-hydroxylase increases bile acid pool size despite decreased enzyme activity. AB - Dietary cholesterol regulation of cholesterol 7alpha-hydroxylase (Cyp7a1), the rate-limiting enzyme in the classical pathway of bile acid synthesis, has been implicated in plasma cholesterol responsiveness. In the current study, the effects of 0.0% and 0.5% cholesterol diets were examined in Cyp7a1 knockout (KO), heterozygous Cyp7a1 KO (Het), and human Cyp7a1 transgenic mice on the mouse Cyp7a1 KO background (Tg+KO). We confirmed previous findings that dietary cholesterol increased mouse Cyp7a1 activity in Het mice but decreased human Cyp7a1 activity in Tg+KO mice. However, in both Het and Tg+KO mice, dietary cholesterol increased bile acid pool size (36% and 72%, respectively) and fecal bile acid excretion (2.2- and 3.6-fold, respectively). The expression of cholesterol 27-hydroxylase (Cyp27), the major enzyme of the alternative pathway of bile acid synthesis, was not significantly different in cholesterol-fed KO, Het, or Tg+KO mice. Furthermore, dietary cholesterol had comparable effects on total plasma cholesterol and non-high-density lipoprotein cholesterol in KO, Het, and Tg+KO mice. Thus, in Tg+KO mice, dietary cholesterol regulates bile acid pool size, fecal bile acid excretion, and plasma cholesterol independently of Cyp7a1 activity. These results challenge the notion that dietary cholesterol regulation of Cyp7a1 is a major determinant of plasma cholesterol responsiveness. PMID- 14762173 TI - Modification of Helicobacter pylori outer membrane protein expression during experimental infection of rhesus macaques. AB - Clinical isolates of Helicobacter pylori show marked diversity, which may derive from genomic changes that occur during the often lifelong association of the bacterium with its human host. We used the rhesus macaque model, together with DNA microarrays, to examine genomic changes in H. pylori that occur early during experimental infection. Microarray analysis showed that H. pylori recovered from challenged macaques had deleted babA, a member of a large family of paralogous outer membrane proteins (OMPs) that mediates attachment of H. pylori to the Lewis B blood group antigen on gastric epithelium. In some cases the babA gene was replaced by babB, an uncharacterized OMP that is closely related to babA. In other cases the babA gene was present but was not expressed because of alteration in dinucleotide CT repeats in the 5' coding region. In either case, strains lacking babA did not adhere to Lewis B, which is expressed on macaque gastric epithelium. Absence of babA and duplication of babB was also seen in H. pylori isolates derived from human clinical samples, suggesting that this gene conversion event is not unique to experimentally infected rhesus monkeys. These results demonstrate in real time with a relevant animal model that H. pylori regulates OMP expression in vivo by using both antigenic variation and phase variation. We suggest that changes in babA and babB after experimental infection of macaques represent a dynamic response in the H. pylori outer membrane that facilitates adherence to the gastric epithelium and promotes chronic infection. PMID- 14762174 TI - A comprehensive nonredundant expressed sequence tag collection for the developing Rattus norvegicus heart. AB - Congenital heart defects affect approximately 1,000,000 people in the United States, with 40,000 new births contributing to that number every year. A large percentage of these defects can be attributed to septal defects. We assembled a nonredundant collection of over 12,000 expressed sequence tags (ESTs) from a total of 30,000 ESTs, with the ultimate goal of identifying spatially and/or temporally regulated genes during heart septation. These ESTs were compiled from nonnormalized, normalized, and serially subtracted cDNA libraries derived from two sets of tissue samples. The first includes microdissected rat hearts from embryonic (E) days E13, E15, and E16.5-E18.5 and adult heart. The second includes hearts from embryonic days E17, E19, and E21 and postnatal (P) days P1, P12, P74, and P200. Over 6,000 novel ESTs were identified in the libraries derived from these two sets of tissues, all of which have been contributed to the NCBI rat UniGene collection. It is anticipated that such EST and cDNA clone resources will prove invaluable to gene expression studies aimed at the understanding of the molecular mechanisms underlying heart septation defects. PMID- 14762175 TI - Starvation response in mouse liver shows strong correlation with life-span prolonging processes. AB - We have monitored global changes in gene expression in mouse liver in response to fasting and sugar-fed conditions using high-density microarrays. From approximately 20,000 different genes, the significantly regulated ones were grouped into specific signaling and metabolic pathways. Striking changes in lipid signaling cascade, insulin and dehydroepiandrosterone (DHEA) hormonal pathways, urea cycle and S-adenosylmethionine-based methyl transfer systems, and cell apoptosis regulators were observed. Since these pathways have been implicated to play a role in the aging process, and since we observe significant overlap of genes regulated upon starvation with those regulated upon caloric restriction, our analysis suggests that starvation may elicit a stress response that is also elicited during caloric restriction. Therefore, many of the signaling and metabolic components regulated during fasting may be the same as those which mediate caloric restriction-dependent life-span extension. PMID- 14762176 TI - Molecular heterogeneity of calcium channel beta-subunits in canine and human heart: evidence for differential subcellular localization. AB - Multiple Ca2+ channel beta-subunit (Ca(v)beta) isoforms are known to differentially regulate the functional properties and membrane trafficking of high-voltage-activated Ca2+ channels, but the precise isoform expression pattern of Ca(v)beta subunits in ventricular muscle has not been fully characterized. Using sequence data from the Human Genome Project to define the intron/exon structure of the four known Ca(v)beta genes, we designed a systematic RT-PCR strategy to screen human and canine left ventricular myocardial samples for all known Ca(v)beta isoforms. A total of 18 different Ca(v)beta isoforms were detected in both canine and human ventricles including splice variants from all four Ca(v)beta genes. Six of these isoforms have not previously been described. Western blots of ventricular membrane fractions and immunocytochemistry demonstrated that all four Ca(v)beta subunit genes are expressed at the protein level, and the Ca(v)beta subunits show differential subcellular localization with Ca(v)beta1b, Ca(v)beta2, and Ca(v)beta3 predominantly localized to the T-tubule sarcolemma, whereas Ca(v)beta1a and Ca(v)beta4 are more prevalent in the surface sarcolemma. Coexpression of the novel Ca(v)beta2c subunits (Ca(v)beta(2cN1), Ca(v)beta(2cN2), Ca(v)beta(2cN4)) with the pore-forming alpha1C (Ca(v)1.2) and Ca(v)alpha2delta subunits in HEK 293 cells resulted in a marked increase in ionic current and Ca(v)beta2c isoform-specific modulation of voltage-dependent activation. These results demonstrate a previously unappreciated heterogeneity of Ca(v)beta subunit isoforms in ventricular myocytes and suggest the presence of different subcellular populations of Ca2+ channels with distinct functional properties. PMID- 14762177 TI - Weighing optimal graft survival through HLA matching against the equitable distribution of kidney allografts. PMID- 14762178 TI - Black mornings, yellow sunsets--a day with paroxysmal nocturnal hemoglobinuria. PMID- 14762179 TI - Bovine spongiform encephalopathy in the United States--an epidemiologist's view. PMID- 14762180 TI - The artificial heart. PMID- 14762181 TI - Effect of changing the priority for HLA matching on the rates and outcomes of kidney transplantation in minority groups. AB - BACKGROUND: HLA typing and the time a patient has spent on the waiting list are the primary criteria used to allocate cadaveric kidneys for transplantation in the United States. Candidates with no HLA-A, B, and DR mismatches are given top priority, followed by candidates with the fewest mismatches at the HLA-B and DR loci; this policy contributes to a higher transplantation rate among whites than nonwhites. We hypothesized that changing this allocation policy would affect graft survival and the racial balance among transplant recipients. METHODS: We estimated the relative rates of kidney transplantation according to race resulting from the current allocation policy and racial differences in HLA antigen profiles, using a Cox model for the time from placement on the waiting list to transplantation. Another model, also adjusted for HLA-B and DR antigen profiles, estimated the relative rates of kidney transplantation that would result if the distribution of these antigen profiles were identical among the racial and ethnic groups. We also investigated the effect of HLA matching on the risk of graft failure, using a Cox model for the time from the first transplantation to graft failure. The results of the two analyses were used to estimate the change in the racial balance of transplantation and graft-failure rates that would result from the elimination of HLA-B matching or HLA-B and DR matching as a means of assigning priority. RESULTS: Eliminating the HLA-B matching as a priority while maintaining HLA-DR matching as a priority would decrease the number of transplantations among whites by 4.0 percent (166 fewer transplantations over a one-year period), whereas it would increase the number among nonwhites by 6.3 percent and increase the rate of graft loss by 2.0 percent. CONCLUSIONS: Removing HLA-B matching as a priority for the allocation of cadaveric kidneys could reduce the existing racial imbalance by increasing the number of transplantations among nonwhites, with only a small increase in the rate of graft loss. PMID- 14762182 TI - Effect of eculizumab on hemolysis and transfusion requirements in patients with paroxysmal nocturnal hemoglobinuria. AB - BACKGROUND: Paroxysmal nocturnal hemoglobinuria (PNH) arises from a somatic mutation of the PIG-A gene in a hematopoietic stem cell and the subsequent production of blood cells with a deficiency of surface proteins that protect the cells against attack by the complement system. We tested the clinical efficacy of eculizumab, a humanized antibody that inhibits the activation of terminal complement components, in patients with PNH. METHODS: Eleven transfusion dependent patients with PNH received infusions of eculizumab (600 mg) every week for four weeks, followed one week later by a 900-mg dose and then by 900 mg every other week through week 12. Clinical and biochemical indicators of hemolysis were measured throughout the trial. RESULTS: Mean lactate dehydrogenase levels decreased from 3111 IU per liter before treatment to 594 IU per liter during treatment (P=0.002). The mean percentage of PNH type III erythrocytes increased from 36.7 percent of the total erythrocyte population to 59.2 percent (P=0.005). The mean and median transfusion rates decreased from 2.1 and 1.8 units per patient per month to 0.6 and 0.0 units per patient per month, respectively (P=0.003 for the comparison of the median rates). Episodes of hemoglobinuria were reduced by 96 percent (P<0.001), and measurements of the quality of life improved significantly. CONCLUSIONS: Eculizumab is safe and well tolerated in patients with PNH. This antibody against terminal complement protein C5 reduces intravascular hemolysis, hemoglobinuria, and the need for transfusion, with an associated improvement in the quality of life in patients with PNH. PMID- 14762183 TI - Association of cystic fibrosis with abnormalities in fatty acid metabolism. AB - BACKGROUND: Patients with cystic fibrosis have altered levels of plasma fatty acids. We previously demonstrated that arachidonic acid levels are increased and docosahexaenoic acid levels are decreased in affected tissues from cystic fibrosis-knockout mice. In this study we determined whether humans with mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene have a similar fatty acid defect in tissues expressing CFTR. METHODS: Fatty acids from nasal- and rectal-biopsy specimens, nasal epithelial scrapings, and plasma were analyzed from 38 subjects with cystic fibrosis and compared with results in 13 obligate heterozygotes, 24 healthy controls, 11 subjects with inflammatory bowel disease, 9 subjects with upper respiratory tract infection, and 16 subjects with asthma. RESULTS: The ratio of arachidonic to docosahexaenoic acid was increased in mucosal and submucosal nasal-biopsy specimens (P<0.001) and rectal-biopsy specimens (P=0.009) from subjects with cystic fibrosis and pancreatic sufficiency and subjects with cystic fibrosis and pancreatic insufficiency, as compared with values in healthy control subjects. In nasal tissue, this change reflected an increase in arachidonic acid levels and a decrease in docosahexaenoic acid levels. In cells from nasal mucosa, the ratio of arachidonic to docosahexaenoic acid was increased in subjects with cystic fibrosis (P<0.001), as compared with healthy controls, with values in obligate heterozygotes intermediate between these two groups (P<0.001). The ratio was not increased in subjects with inflammatory bowel disease. Subjects with asthma and those with upper respiratory tract infection had values intermediate between those in subjects with cystic fibrosis and those in healthy control subjects. CONCLUSIONS: These data indicate that alterations in fatty acids similar to those in cystic fibrosis-knockout mice are present in CFTR-expressing tissue from subjects with cystic fibrosis. PMID- 14762184 TI - Deficiency of the circulating insulin-like growth factor system associated with inactivation of the acid-labile subunit gene. PMID- 14762186 TI - Parvovirus B19. PMID- 14762185 TI - The Budd-Chiari syndrome. PMID- 14762187 TI - Images in clinical medicine. Parvovirus B19 infection. PMID- 14762188 TI - Clinical problem-solving. Occam's razor versus Saint's Triad. PMID- 14762189 TI - Fatty acid metabolism in cystic fibrosis. PMID- 14762190 TI - The pharmaceutical industry versus Medicaid--limits on state initiatives to control prescription-drug costs. PMID- 14762191 TI - Getting a handle on graft-versus-host disease. PMID- 14762192 TI - Ximelagatran versus warfarin after total knee replacement. PMID- 14762193 TI - Ximelagatran for secondary prevention of venous thromboembolism. PMID- 14762194 TI - First-trimester screening for Down's syndrome. PMID- 14762195 TI - Fondaparinux in pulmonary embolism. PMID- 14762196 TI - Effects of estrogen plus progestin on health-related quality of life. PMID- 14762197 TI - Potts shunt in patients with pulmonary hypertension. PMID- 14762198 TI - Tissue matching before the era of HLA typing. PMID- 14762199 TI - Images in clinical medicine. Congenital lymphedema. PMID- 14762200 TI - Two-dimensional conformation-dependent electrophoresis (2D-CDE) to separate DNA fragments containing unmatched bulge from complex DNA samples. AB - DNA fragments containing mispaired and modified bases, bulges, lesions and specific sequences have altered conformation. Methods for separating complex samples of DNA fragments based on conformation but independent of length have many applications, including (i) separation of mismatched or unmatched DNA fragments from those perfectly matched; (ii) simultaneous, diagnostic, mismatch scanning of multiple fragments; (iii) isolation of damaged DNA fragments from undamaged fragments; and (iv) estimation of reannealing efficiency of complex DNA samples. We developed a two-dimensional conformation-dependent electrophoresis (2D-CDE) method for separating DNA fragments based on length and conformation in the first dimension and only on length in the second dimension. Differences in migration velocity due to conformation were minimized during second dimension electrophoresis by introducing an intercalator. To test the method, we constructed 298 bp DNA fragments containing cytosine bulges ranging from 1 to 5 nt. Bulge-containing DNA fragments had reduced migration velocity in the first dimension due to altered conformation. After 2D-CDE, bulge-containing DNA fragments had migrated in front of an arc comprising heterogeneous fragments with regular conformation. This simple and robust method could be used in both analytical and preparative applications involving complex DNA samples. PMID- 14762201 TI - Effects on RNAi of the tight structure, sequence and position of the targeted region. AB - RNA interference (RNAi) is a gene-silencing phenomenon that involves the double stranded RNA-mediated cleavage of mRNA, and small interfering RNAs (siRNAs) can cause RNAi in mammalian cells. There have been many attempts to clarify the mechanism of RNAi, but information about the relationship between the sequence and structure, in particular, a tight structure, of the target RNA and the activities of siRNAs are limited. In the present study, we examined this relationship by introducing the TAR element, which adopts a very stable secondary structure, at different positions within target RNAs. Our results suggested that the activities of siRNAs were affected by the tight stem-loop structure of TAR. In contrast, the position of the target within the mRNA, the binding of the Tat protein to the TAR, and the location of the target within a translated or a noncoding region had only marginal effects on RNAi. When the target sequence was placed in two different orientations, only one orientation had a significant effect on the activities of siRNA, demonstrating that the presence of certain nucleotides at some specific positions was favorable for RNAi. Systematic analysis of 47 different sites within 47 plasmids under identical conditions indicated that it is the target sequence itself, rather than its location, that is the major determinant of siRNA activity. PMID- 14762202 TI - High-throughput protein analysis integrating bioinformatics and experimental assays. AB - The wealth of transcript information that has been made publicly available in recent years requires the development of high-throughput functional genomics and proteomics approaches for its analysis. Such approaches need suitable data integration procedures and a high level of automation in order to gain maximum benefit from the results generated. We have designed an automatic pipeline to analyse annotated open reading frames (ORFs) stemming from full-length cDNAs produced mainly by the German cDNA Consortium. The ORFs are cloned into expression vectors for use in large-scale assays such as the determination of subcellular protein localization or kinase reaction specificity. Additionally, all identified ORFs undergo exhaustive bioinformatic analysis such as similarity searches, protein domain architecture determination and prediction of physicochemical characteristics and secondary structure, using a wide variety of bioinformatic methods in combination with the most up-to-date public databases (e.g. PRINTS, BLOCKS, INTERPRO, PROSITE SWISSPROT). Data from experimental results and from the bioinformatic analysis are integrated and stored in a relational database (MS SQL-Server), which makes it possible for researchers to find answers to biological questions easily, thereby speeding up the selection of targets for further analysis. The designed pipeline constitutes a new automatic approach to obtaining and administrating relevant biological data from high throughput investigations of cDNAs in order to systematically identify and characterize novel genes, as well as to comprehensively describe the function of the encoded proteins. PMID- 14762203 TI - Transcription regulation of TNF-alpha-early response genes by poly(ADP-ribose) polymerase-1 in murine heart endothelial cells. AB - Poly(ADP-ribose) polymerase-1 (PARP-1) has been involved in endothelial cell dysfunction associated with various pathophysiological conditions. The intrinsic mechanism of PARP-1-mediated endothelial cell dysfunction could be related to PARP-1 overactivation, NAD(+) consumption and ATP depletion. An alternative way could involve transcription regulation. By using high-density microarrays, we examined early tumor necrosis factor alpha (TNF-alpha)-stimulated gene expression profiles in PARP-1(+/+) and PARP-1(-/-) murine heart endothelial cells. TNF-alpha modulated a significant number of genes in both cell types. We have identified a set of genes whose expression in response to TNF-alpha is modulated by PARP-1, whereas the expression of others is PARP-1-independent. Up-regulation of several genes involved in the inflammatory response is hampered in the absence of PARP-1. Moreover, NF-kappaB-dependent transcriptional activation is partially inhibited in PARP-1(-/-) compared to PARP-1(+/+) cells. However, we found that PARP-1 might also silence transcription of several NF-kappaB target genes. Overall, our results show that PARP-1 is regulating the expression of genes by the endothelial cells both in a positive and a negative fashion, with the final effects depending on the gene. Individual studies of these genes are now necessary to clarify the intrinsic mechanism by which PARP-1 is controlling transcription and thereby finding out different therapeutic approaches involving PARP-1. PMID- 14762204 TI - Involvement of Hus1 in the chain elongation step of DNA replication after exposure to camptothecin or ionizing radiation. AB - DNA damage-induced S phase (S) checkpoint includes inhibition of both replicon initiation and chain elongation. The precise mechanism for controlling the two processes remains unclear. In this study, we showed that Hus1-deficient mouse cells had an impaired S checkpoint after exposure to DNA strand break-inducing agents such as camptothecin (CPT) (>or=1.0 micro M), or ionizing radiation (IR) (>or=15 Gy). The Hus1-dependent S checkpoint contributes to cell resistance to CPT. This impaired S checkpoint induced by CPT or IR in Hus1-deficient cells reflected mainly the chain elongation step of DNA replication and was correlated with the reduction of dissociation of PCNA from DNA replication foci. Although Hus1 is required for Rad9 phosphorylation following exposure of cells to CPT or IR, Hus1-deficient cells showed normal activation of ATR/CHK1 and ATM kinases at doses where the checkpoint defects were manifested, suggesting that Hus1 is not a component of the sensor system for activating these pathways in S checkpoint induced by CPT or IR. PMID- 14762205 TI - A programmed -1 ribosomal frameshift signal can function as a cis-acting mRNA destabilizing element. AB - Nonsense-mediated mRNA decay (NMD) directs rapid degradation of premature termination codon (PTC)-containing mRNAs, e.g. those containing frameshift mutations. Many viral mRNAs encode polycistronic messages where programmed -1 ribosomal frameshift (-1 PRF) signals direct ribosomes to synthesize polyproteins. A previous study, which identified consensus -1 PRF signals in the yeast genome, found that, in contrast to viruses, the majority of predicted -1 PRF events would direct translating ribosomes to PTCs. Here we tested the hypothesis that a -1 PRF signal can function as a cis-acting mRNA destabilizing element by inserting an L-A viral -1 PRF signal into a PGK1 reporter construct in the 'genomic' orientation. The results show that even low levels of -1 PRF are sufficient to target the reporter mRNA for degradation via the NMD pathway, with half-lives similar to messages containing in-frame PTCs. The demonstration of an inverse correlation between frameshift efficiency and mRNA half-lives suggests that modulation of -1 PRF frequencies can be used to post-transcriptionally regulate gene expression. Analysis of the mRNA decay profiles of the frameshift signal- containing reporter mRNAs also supports the notion that NMD remains active on mRNAs beyond the 'pioneer round' of translation in yeast. PMID- 14762206 TI - VITO-1 is an essential cofactor of TEF1-dependent muscle-specific gene regulation. AB - The expression of several muscle-specific genes is partially or completely regulated by MCAT elements, which bind members of the TEF family of transcription factors. TEF1 itself is unable to activate reporter plasmids bearing TEF1-binding sites, suggesting that additional bridging or co-activating factors are necessary to allow interaction of TEF1 with the transcriptional machinery. In addition, none of the known TEF genes are exclusively expressed in the cardiac or skeletal muscle lineage to account for the muscle-specific expression of MCAT-dependent genes. Here we describe that VITO-1, a new SID (scalloped interaction domain) containing protein, binds to TEF1 in vitro and strongly stimulates transcription of a MCAT reporter plasmid together with TEF-1. Since VITO-1 is predominantly expressed in the skeletal muscle lineage, it might serve as an essential transcriptional intermediary factor to promote muscle-specific expression via MCAT cis-regulatory elements. Although VITO-1 alone is not sufficient to initiate myogenic conversion of 10T1/2 fibroblastic cells, it enhanced MyoD-mediated myogenic conversion. In addition, interference with VITO-1 expression by siRNA attenuated differentiation of C2C12 muscle cells and MyoD-dependent myogenesis in 10T1/2 cells. We conclude that VITO-1 is a crucial new cofactor of the muscle regulatory programme. PMID- 14762207 TI - Recent horizontal intron transfer to a chloroplast genome. AB - Evidence is presented for the recent, horizontal transfer of a self-splicing, homing group II intron from a cyanobacteria to the chloroplast genome of Euglena myxocylindracea. The psbA gene of E.myxocylindracea was found to contain a single 2566 nt group II intron with a gene in domain 4 for a 575 amino acid maturase. The predicted secondary structure and tertiary interactions of the group II intron, as well as the derived maturase primary sequence, most closely resemble the homing intron of the cyanobacterium Calothrix and the rnl introns of Porphyra purpurea mitochondria, while being only distantly related to all other Euglena plastid introns and maturases. All main functional domains of the intron-encoded proteins of known homing introns are conserved, including reverse transcriptase domains 1-7, the zinc finger domain and domain X. The close relationship with cyanobacterial introns was confirmed by phylogenetic analysis. Both the full length psbA intron and a Delta-maturase variant self-splice in vitro in two independent assays. The psbA intron is the first example of a self-splicing chloroplast group II intron from any organism. These results support the conclusion that the psbA intron is the result of a recent horizontal transfer into the E.myxocylindracea chloroplast genome from a cyanobacterial donor and should prompt a reconsideration of horizontal transfer mechanisms to account for the origin of other chloroplast genetic elements. PMID- 14762208 TI - The sea urchin stem-loop-binding protein: a maternally expressed protein that probably functions in expression of multiple classes of histone mRNA. AB - Following the completion of oogenesis and oocyte maturation, histone mRNAs are synthesized and stored in the sea urchin egg pronucleus. Histone mRNAs are the only mRNAs that are not polyadenylated but instead end in a stem-loop which has been conserved in evolution. The 3' end binds the stem-loop-binding protein (SLBP), and SLBP is required for histone pre-mRNA processing as well as translation of the histone mRNAs. A cDNA encoding a 59 kDa sea urchin SLBP (suSLBP) has been cloned from an oocyte cDNA library. The suSLBP contains an RNA binding domain that is similar to the RNA-binding domain found in SLBPs from other species, although there is no similarity between the rest of the suSLBP and other SLBPs. The suSLBP is present at constant levels in eggs and for the first 12 h of development. The levels of suSLBP then decline and remain at a low level for the rest of embryogenesis. The suSLBP is concentrated in the egg pronucleus and is released from the nucleus only when cells enter the first mitosis. SuSLBP expressed by in vitro translation does not bind the stem-loop RNA, suggesting that suSLBP is modified to activate RNA binding in sea urchin embryos. PMID- 14762209 TI - Probing alternative foldings of the HIV-1 leader RNA by antisense oligonucleotide scanning arrays. AB - Scanning arrays of antisense DNA oligonucleotides provide a novel and systematic means to study structural features within an RNA molecule. We used this approach to probe the structure of the untranslated leader of the human immunodeficiency virus type 1 (HIV-1) RNA genome. This 335 nt RNA encodes multiple important replication signals and adopts two mutually exclusive conformations. The poly(A) and the dimer initiation signal (DIS) sequences of the leader RNA are base-paired in the long-distance interaction (LDI) conformation, but both domains form distinct hairpins in the branched multiple hairpins (BMH) conformation. An RNA switch mechanism has been proposed to regulate the activity of the DIS dimerization signal that is masked in one, yet exposed in the other conformation. The two RNA conformations demonstrate discrete differences in the array-based hybridization patterns. LDI shows increased hybridization in the poly(A) region and decreased hybridization in the DIS region when compared with BMH. These results provide additional evidence for the structure models of the two alternative leader RNA conformations. We also found a correlation between the efficiency of oligonucleotide hybridization and the accessibility of the RNA structure as determined by chemical and enzymatic probing in previous studies. The array approach therefore provides a very sensitive method to detect structural differences in related transcripts. PMID- 14762210 TI - 2'-O-[2-[(N,N-dimethylamino)oxy]ethyl]-modified oligonucleotides inhibit expression of mRNA in vitro and in vivo. AB - Synthesis and antisense activity of oligonucleotides modified with 2'-O-[2-[(N,N dimethylamino)oxy] ethyl] (2'-O-DMAOE) are described. The 2'-O-DMAOE-modified oligonucleotides showed superior metabolic stability in mice. The phosphorothioate oligonucleotide 'gapmers', with 2'-O-DMAOE- modified nucleoside residues at the ends and 2'-deoxy nucleosides residues in the central region, showed dose-dependent inhibition of mRNA expression in cell culture for two targets. 'Gapmer' oligonucleotides have one or two 2'-O-modified regions and a 2' deoxyoligonucleotide phosphorothioate region that allows RNase H digestion of target mRNA. To determine the in vivo potency and efficacy, BalbC mice were treated with 2'-O-DMAOE gapmers and a dose-dependent reduction in the targeted C raf mRNA expression was observed. Oligonucleotides with 2'-O-DMAOE modifications throughout the sequences reduced the intercellular adhesion molecule-1 (ICAM-1) protein expression very efficiently in HUVEC cells with an IC(50) of 1.8 nM. The inhibition of ICAM-1 protein expression by these uniformly modified 2'-O-DMAOE oligonucleotides may be due to selective interference with the formation of the translational initiation complex. These results demonstrate that 2'-O-DMAOE- modified oligonucleotides are useful for antisense-based therapeutics when either RNase H-dependent or RNase H-independent target reduction mechanisms are employed. PMID- 14762211 TI - Role of the RNA polymerase alpha subunits in CII-dependent activation of the bacteriophage lambda pE promoter: identification of important residues and positioning of the alpha C-terminal domains. AB - The bacteriophage lambda CII protein stimulates the activity of three phage promoters, p(E), p(I) and p(aQ), upon binding to a site overlapping the -35 element at each promoter. Here we used preparations of RNA polymerase carrying a DNA cleavage reagent attached to specific residues in the C-terminal domain of the RNA polymerase alpha subunit (alphaCTD) to demonstrate that one alphaCTD binds near position -41 at p(E), whilst the other alphaCTD binds further upstream. The alphaCTD bound near position -41 is oriented such that its 261 determinant is in close proximity to sigma(70). The location of alphaCTD in CII dependent complexes at the p(E) promoter is very similar to that found at many activator-independent promoters, and represents an alternative configuration for alphaCTD at promoters where activators bind sites overlapping the -35 region. We also used an in vivo alanine scan analysis to show that the DNA-binding determinant of alphaCTD is involved in stimulation of the p(E) promoter by CII, and this was confirmed by in vitro transcription assays. We also show that whereas the K271E substitution in alphaCTD results in a drastic decrease in CII dependent activation of p(E), the p(I) and p(aQ) promoters are less sensitive to this substitution, suggesting that the role of alphaCTD at the three lysogenic promoters may be different. PMID- 14762212 TI - Disruption of type III secretion in Salmonella enterica serovar Typhimurium by external guide sequences. AB - The type III secretion system involved in Salmonella enterica serovar Typhimurium invasion of host cells has been disrupted using inducibly expressed oligonucleotide external guide sequences (EGSs) complementary to invB or invC mRNA. These EGSs direct single site cleavage in these mRNAs by endogenous RNase P, and their expression in Salmonella results in invC mRNA and InvC protein depletion, decreased type III secretion and interference with host cell invasion. Comparison of these effects with those from studies of Salmonella invB and invC mutants suggests that invB EGSs have polar effects on invC mRNA. PMID- 14762214 TI - Improved detection of hydrophilic phosphopeptides using graphite powder microcolumns and mass spectrometry: evidence for in vivo doubly phosphorylated dynamin I and dynamin III. AB - A common strategy in proteomics to improve the number and quality of peptides detected by mass spectrometry (MS) is to desalt and concentrate proteolytic digests using reversed phase (RP) chromatography prior to analysis. However, this does not allow for detection of small or hydrophilic peptides, or peptides altered in hydrophilicity such as phosphopeptides. We used microcolumns to compare the ability of RP resin or graphite powder to retain phosphopeptides. A number of standard phosphopeptides and a biologically relevant phosphoprotein, dynamin I, were analyzed. MS revealed that some phosphopeptides did not bind the RP resin but were retained efficiently on the graphite. Those that did bind the RP resin often produced much stronger signals from the graphite powder. In particular, the method revealed a doubly phosphorylated peptide in a tryptic digest of dynamin I purified from rat brain nerve terminals. The detection of this peptide was greatly enhanced by graphite micropurification. Sequencing by tandem MS confirmed the presence of phosphate at both Ser-774 and Ser-778, while a singly phosphorylated peptide was predominantly phosphorylated only on Ser-774. The method further revealed a singly and doubly phosphorylated peptide in dynamin III, analogous to the dynamin I sequence. A pair of dynamin III phosphorylation sites were found at Ser-759 and Ser-763 by tandem MS. The results directly define the in vivo phosphorylation sites in dynamins I and III for the first time. The findings indicate a large improvement in the detection of small amounts of phosphopeptides by MS and the approach has major implications for both small- and large-scale projects in phosphoproteomics. PMID- 14762213 TI - Fission yeast global repressors regulate the specificity of chromatin alteration in response to distinct environmental stresses. AB - The specific induction of genes in response to distinct environmental stress is vital for all eukaryotes. To study the mechanisms that result in selective gene responses, we examined the role of the fission yeast Tup1 family repressors in chromatin regulation. We found that chromatin structure around a cAMP-responsive element (CRE)-like sequence in ade6-M26 that is bound by Atf1.Pcr1 transcriptional activation was altered in response to osmotic stress but not to heat and oxidative stresses. Such chromatin structure alteration occurred later than the Atf1 phosphorylation but correlated well with stress-induced transcriptional activation at ade6-M26. This chromatin structure alteration required components for the stress-activated protein kinase (SAPK) cascade and both subunits of the M26-binding CREB/ATF-type protein Atf1.Pcr1. Cation stress and glucose starvation selectively caused chromatin structure alteration around CRE-like sequences in cta3(+) and fbp1(+) promoters, respectively, in correlation with transcriptional activation. However, the tup11Delta tup12Delta double deletion mutants lost the selectivity of stress responses of chromatin structure and transcriptional regulation of cta3(+) and fbp1(+). These data indicate that the Tup1-like repressors regulate the chromatin structure to ensure the specificity of gene activation in response to particular stresses. Such a role for these proteins may serve as a paradigm for the regulation of stress response in higher eukaryotes. PMID- 14762215 TI - Targeted therapy in breast cancer: the HER-2/neu gene and protein. AB - The HER-2/neu oncogene, a member of the epidermal growth factor receptor or erb gene family, encodes a transmembrane tyrosine kinase receptor that has been linked to prognosis and response to therapy with the anti-HER-2-humanized monoclonal antibody, trastuzumab (Herceptin, Genentech, South San Francisco, CA) in patients with advanced metastatic breast cancer. HER-2/neu status has also been tested for its ability to predict the response of breast cancer to other therapies including hormonal therapies, topoisomerase inhibitors, and anthracyclines. This review includes an analysis of 80 published studies encompassing more than 25,000 patients designed to consider the relative advantages and disadvantages of the various methods of measuring HER-2/neu in clinical breast cancer specimens. Southern blotting, PCR amplification detection, and fluorescence in situ hybridization assays designed to detect HER-2/neu gene amplification are compared with HER-2/neu protein overexpression assays performed by immunohistochemical techniques applied to frozen and paraffin-embedded tissues and enzyme immunoassays performed on tumor cytosols. The significance of HER 2/neu overexpression in ductal carcinoma in situ and the HER-2/neu status in uncommon female breast conditions and male breast cancer are also considered. The role of HER-2/neu testing for the prediction of response to trastuzumab therapy in breast cancer is reviewed along with the current studies designed to test whether HER-2/neu status can predict the response to standard and newer hormonal therapies, cytotoxic chemotherapy, and radiation. The review will also evaluate the status of serum-based testing for circulating HER-2/neu receptor protein and its ability to predict disease outcome and therapy response. PMID- 14762216 TI - Clinical infrastructures to support proteomic studies of tissue and fluids in breast cancer. AB - The Danish Breast Cancer Cooperative Group (DBCG) was established in 1977 with the aim to ensure optimal breast cancer diagnostics and therapeutic modalities on a nationwide basis. DBCG was organized in such a way so it represents a broad interdisciplinary collaboration with established clinical databases and biobanks. This review summarizes the infrastructures, such as those of the DBCG, that are required to facilitate translational research studies aiming at further diagnostic and therapeutic improvements through interactions directed at prevention, early diagnosis, and treatment of primary breast cancer. PMID- 14762217 TI - Stimulation of cells using EGF-coated magnetic beads. AB - The ability to determine the spatial localization of signals can assist in defining the regulation of specific signaling pathways within the cell. Localized application of a stimulus, followed by imaging of various proteins or their phosphorylated products, allows visualization of the propagation scales of responses to the stimulus. To obtain a localized stimulus of epidermal growth factor (EGF), we bound the ligand to magnetic beads, which could be rapidly pulled onto cell surfaces by a magnet. This method allows the spatial analysis of EGF-induced signaling pathways. PMID- 14762218 TI - G protein signaling: insights from new structures. AB - A large body of experimental evidence exists that links heterotrimeric guanosine triphosphate-binding protein (G protein) structure to function. The determination of the crystal structures of G proteins in various activational states and, more recently, in complexes with effectors and other signaling partners highlights the varied mechanisms involved in G protein regulation. Signaling complexes, such as the recently solved complex of Gbetagamma and G protein receptor kinase 2 (GRK2), provide new insights into the mechanisms underlying the regulation of these highly conserved signaling molecules. In this Review, we discuss the latest findings and their implications for G protein-signaling paradigms. PMID- 14762220 TI - Placing bets. National Institute on Aging committee chooses three compounds for definitive longevity testing. PMID- 14762221 TI - Tugging at heartstrings. Protein associated with heart abnormalities and premature aging syndrome transmits mechanical stress to the nucleus. PMID- 14762223 TI - Detection of Pneumocystis carinii by DNA amplification in patients with connective tissue diseases: re-evaluation of clinical features of P. carinii pneumonia in rheumatic diseases. AB - OBJECTIVES: We evaluated the polymerase chain reaction (PCR) detection of Pneumocystis carinii DNA in induced sputum of patients with connective tissue diseases and assessed the clinical features of patients positive for P. carinii. METHODS: Sputum was induced by nebulization in 29 in-patients with various connective tissue diseases who presented with symptoms suggestive of P. carinii pneumonia (PCP), and was examined by PCR. RESULTS: Detection of P. carinii DNA by PCR was significantly more sensitive than cytology; 54.5% patients were positive by PCR and only 4.5% by cytology. The prevalence of PCP was higher than previously considered and was especially high in patients receiving > 30 mg/day prednisolone with or without other immunosuppressants. P. carinii-positive patients had significant lymphocytopenia and a low serum IgG level compared with P. carinii-negative patients. P. carinii disappeared within 7-10 days after therapy with trimethoprim/sulfamethoxazole. CONCLUSION: We propose that the use of PCR for detection of P. carinii using induced sputum is a useful and non invasive method that has high sensitivity and specificity for the early diagnosis of PCP. PMID- 14762222 TI - From genes to societies. AB - Research on model organisms has substantially advanced our understanding of aging. However, these studies collectively lack any examination of the element of sociality, an important feature of human biology. Social insects present a number of unique possibilities for investigating social influences on aging and potentially detecting new mechanisms for extremely prolonged, healthy life spans that have evolved naturally. Social evolution has led to life spans in reproductive females that are much longer (up to over 100-fold) than those of males or of nonreproductive worker castes. These differences are particularly dramatic because they are due to environmental influences, as all individuals develop from the same genomes. Social insect colonies consist of semi-autonomous individuals, and the relationship between the colony and the individual creates many interesting predictions in the light of the common theories of aging. Furthermore, the variety of lifestyles of social insects creates the potential for crucial comparative analyses across distinct social systems. PMID- 14762224 TI - Comparison of the activities of multinucleated bone-resorbing giant cells derived from CD14-positive cells in the synovial fluids of rheumatoid arthritis and osteoarthritis patients. AB - OBJECTIVE: To investigate the morphology and function of multinucleated bone resorbing giant cells derived from CD14-positive cells in the synovial fluids (SF) of patients with rheumatoid arthritis (RA) or osteoarthritis (OA). METHODS: CD14-positive cells were obtained by magnetic-activated cell sorting of primary cultures of mononuclear cells from the SF. Multinucleated bone-resorbing giant cells were induced from the CD14-positive cells in the presence or absence of cytokines. We examined various characteristics, including osteoclast markers, fusion index and bone-resorption activities of the multinucleated giant cells. RESULTS: Multinucleated giant cells were induced from the CD14-positive cells in the SF of the RA and OA patients by the addition of interleukin (IL)-3, IL-5 and IL-7, or granulocyte-macrophage colony-stimulating factor (GM-CSF), respectively. These multinucleated giant cells were positive for tartrate-resistant acid phosphatase (TRAP), carbonic anhydrase II, actin, vitronectin receptor and the calcitonin receptor. However, the average values for the number of nuclei, fusion index and bone-resorption functions of the SF cells from the RA patients were significantly higher than those derived from the OA patients. CONCLUSION: These results suggest that the induction and activities of multinucleated bone resorbing giant cells may play a pivotal role in bone destruction, and that these processes may be enhanced significantly in RA patients. PMID- 14762225 TI - Epidemiology of hip and knee pain and its impact on overall health status in older adults. AB - OBJECTIVES: To obtain prevalence rates of hip and knee pain in elderly people and compare combinations of symptoms with overall health status. METHODS: We performed a cross-sectional postal survey of a random sample of 5500 Oxfordshire residents aged 65 yr and older. Prevalence estimates were based on the screening question: 'During the past 12 months, have you had pain in or around either of your hip/knee joints on most days for one month or longer?' Overall health status was assessed with the SF-36 questionnaire. RESULTS: The response rate was 66.3% (3341/5039 eligible people), and was highest (approximately reverse similar 72%) for the 65-74 yr age-group. The percentage reporting hip pain was 19.2% [95% confidence interval (CI) 17.9-20.6], and 32.6% (95% CI 31.0-34.3) reported knee pain. The percentage reporting hip and knee pain was 11.3%, and 40.7% reported hip or knee pain. Less than half (48%) of the symptomatic respondents had unilateral problems affecting one hip or knee joint only. SF-36 scores worsened as the number of symptomatic hip and knee joints increased (P<0.001 for physical function, physical role limitation and bodily pain). CONCLUSIONS: Patterns of hip and knee symptoms are complex in older people. Amongst the symptomatic, most have more than one hip/knee affected. This has implications for treatment and health status measurement. In the absence of hip and knee symptoms, general health status scores of elderly people are similar to those of people aged under 65 yr. PMID- 14762226 TI - Characterization of plasmacytoid dendritic cells in inflammatory arthritis synovial fluid. AB - OBJECTIVE: To examine the phenotype of dendritic cell subsets in synovial fluid and peripheral blood from patients with rheumatoid arthritis (RA) or spondyloarthropathy (SpA). METHODS: Multiparameter flow cytometry was used to identify and characterize dendritic cells in mononuclear cell populations isolated from synovial fluid and peripheral blood. RESULTS: Synovial fluid contained two subsets of dendritic cells (DC), myeloid and plasmacytoid. These subsets could also be identified in peripheral blood, but there were lower numbers of DC in peripheral blood compared with synovial fluid. Plasmacytoid DC were distinguished from the myeloid subset by high expression of CD123 and lack of expression of CD11c. In comparison with myeloid dendritic cells, the plasmacytoid subset were less mature, similar to those in peripheral blood. They failed to express CD83 and DC-LAMP, and had relatively low levels of CD40 and CD86. Comparison of dendritic cells in synovial fluid from RA and SpA patients showed increased numbers of the plasmacytoid subset in SpA. CONCLUSIONS: This is the first demonstration of the plasmacytoid subset of dendritic cells in synovial fluid. Since these cells are major producers of type I interferons, their increased numbers in SpA might be relevant to pathogenesis, but the immature phenotype in SpA synovial fluid may also indicate that conditions for maturation of this subset do not pertain in SpA synovium. PMID- 14762227 TI - Celecoxib 200 mg q.d. is efficacious in the management of osteoarthritis of the knee or hip regardless of the time of dosing. AB - OBJECTIVES: The primary objective was to demonstrate equivalence between a.m. and p.m. dosing of celecoxib 200 mg q.d. An equivalence assessment of q.d. vs b.i.d. dosing was a secondary objective. METHODS: In this randomized, double-blind study, patients with symptomatic osteoarthritis of the knee or hip were randomized to receive celecoxib 200 mg q.d. a.m., celecoxib 200 mg q.d. p.m. or celecoxib 100 mg b.i.d. The primary outcome variable, measured at week 12 on a 0- to 10-point integrated scale, was patient satisfaction assessment (pain relief, walking/bending, and willingness to continue medication). Equivalence was declared if the 95% confidence interval (CI) of the difference (a.m. q.d. vs p.m. q.d., b.i.d. vs q.d.) fell within the interval of -2 to +2. RESULTS: A total of 697 patients were enrolled in this trial. For the a.m. vs p.m. comparison, the 95% CIs were within the prespecified equivalence criteria for all three measures of patient satisfaction: pain relief, mean -0.2, 95% CI -0.53 to 0.68; ability to walk and bend, mean -0.2, 95% CI -0.54 to 0.64; willingness to continue medication, mean -0.7, 95% CI -0.98 to 0.49. The 95% CIs for the q.d. vs b.i.d. comparison were also within the -2 to +2 interval. CONCLUSION: Regardless of the time of day at which celecoxib 200 mg q.d. is administered, patients are equally satisfied with the pain relief, ability to walk and bend, and willingness to continue medication. PMID- 14762228 TI - Patterns of joint pain: lessons from epidemiology. PMID- 14762229 TI - Good idea or politically correct nonsense? PMID- 14762231 TI - Datapoints: adolescents with substance abuse: are health plans missing them? PMID- 14762232 TI - Economic grand rounds: the economic burden of bipolar disorder. PMID- 14762233 TI - Personal accounts: a fragile China doll. PMID- 14762234 TI - Psychopharmacology: atypical antipsychotic dosing: the effect of co-medication with anticonvulsants. PMID- 14762235 TI - Best practices: are financial incentives and best practices compatible? PMID- 14762236 TI - Should patients and their families contribute to the DSM-V process? AB - The authors consider arguments for and against the formal inclusion of mentally ill patients and their families in the deliberative processes leading to DSM-V. These discussions involve six key issues: the scientific status of psychiatric classifications; public policy and political considerations; the practical implications of widening the review process; the capacities of lay members of the workgroups; freedom of expression and the openness of the review process; and the uniqueness of consumer perspectives. The authors conclude that involving patients and families in the DSM review process is supportable on both scientific and public policy grounds. PMID- 14762237 TI - Therapeutic limit setting and six-month outcomes in a Veterans Affairs assertive community treatment program. AB - OBJECTIVE: This study examined the relationship of limit-setting interventions and six-month outcomes in assertive community treatment. METHODS: Case managers from 40 Veterans Affairs assertive community treatment teams at 40 different sites documented their use of 25 limit-setting activities with 1564 clients during the first six months of treatment. Five scales were constructed representing different types of limit-setting activities: withholding certain types of assistance until the client curtailed certain behaviors; behavioral contracting in which specific goals were identified and linked to reinforcers if the goals were achieved; invocation of external authorities, such as a probation officer; seeking a declaration of incompetence to manage funds or initiation of a request for a payee; and forced hospitalization through civil commitment. Structured interviews conducted at baseline and six months documented changes in clinical status and community adjustment. Multiple regression analysis was used to examine the relationship between limit-setting interventions and outcomes at both the level of the individual client and at the level of the team, adjusting for potentially confounding baseline client characteristics. RESULTS: All five measures of specific limit-setting activities were associated with poorer outcomes on four to six of the eight outcome measures. The site-level comparison of outcomes showed more violent behavior at sites that made more extensive use of these interventions but also greater employment. CONCLUSIONS: After the analysis controlled for potentially confounding factors, clients exposed to limit-setting interventions had poorer outcomes than others on many measures, suggesting that within the limits of a nonexperimental study, such interventions do not appear to prevent adverse outcomes. PMID- 14762238 TI - Comparison of offenders with mental illness only and offenders with dual diagnoses. AB - OBJECTIVE: The study compared offenders who had severe mental illness only and offenders who had severe mental illness and substance abuse problems-dual diagnoses-to determine whether these groups differed. Offenders with dual diagnoses who were involved with the criminal justice system at different levels were compared to explore their profiles and experiences after release. METHODS: Secondary data collected on offenders who had diagnoses of severe mental illness and of substance abuse in Massachusetts were used to examine sociodemographic, clinical characteristics, and criminal justice characteristics, service needs, and community reentry experiences in the first three months postrelease of 265 offenders with major mental illness and 436 with dual diagnoses. RESULTS: Offenders with dual diagnoses were more likely to be female and to have a history of being on probation and of using mental health services. On release from correctional custody, they had more immediate service needs than offenders with mental illness alone, including a need for housing and sex offender treatment, and they were more likely to require an assessment for dangerousness. They were also more likely to return to correctional custody. CONCLUSIONS: The data do not suggest that offenders with dual diagnoses have a distinct clinical background, but rather that substance abuse is an important feature that affects their real or perceived level of functioning, engagement with the criminal justice system, and dependence on social service institutions in the community. PMID- 14762239 TI - Ethnicity and prescription patterns for haloperidol, risperidone, and olanzapine. AB - OBJECTIVE: Patients with schizophrenia may respond better to second-generation antipsychotics than to older antipsychotics because of their superior efficacy and safety profiles. However, the reduced likelihood among ethnic minority groups of receiving newer antipsychotics may be associated with reduced medication adherence and health service use, potentially contributing to poor response rates. This study examined whether ethnicity helped predict whether patients with schizophrenia were given a first- or a second-generation antipsychotic, haloperidol versus risperidone or olanzapine, and what type of second-generation antipsychotic was prescribed, risperidone or olanzapine, when other factors were controlled for. METHODS: Texas Medicaid claims were analyzed for persons aged 21 to 65 years with a diagnosis of schizophrenia or schizoaffective disorder who started treatment with olanzapine (N=1875), risperidone (N=982), or haloperidol (N= 726) between January 1, 1997 and August 31, 1998. The association between antipsychotic prescribing patterns among African Americans, Mexican Americans, and whites was assessed by using logistic regression analysis. Covariates included other patient demographic characteristics, region, comorbid mental health conditions, and medication and health care resource use in the 12 months before antipsychotic initiation. RESULTS: The results of the first- versus second generation antipsychotic analysis indicated that African Americans were significantly less likely than whites to receive risperidone or olanzapine. Although not statistically significant, the odds ratio indicated that Mexican Americans were also less likely to receive risperidone or olanzapine. Ethnicity was not associated with significant differences in the prescribing patterns of risperidone versus olanzapine. CONCLUSIONS: When other factors were controlled for, African Americans were significantly less likely to receive the newer antipsychotics. Among those who received the newer antipsychotics, ethnicity did not affect medication choice. PMID- 14762240 TI - Trauma history screening in a community mental health center. AB - OBJECTIVE: This study assessed the lifetime prevalence of traumatic events among consumers of a community mental health center by using a brief trauma screening instrument. This study also examined the relationship between trauma exposure and physical and mental health sequelae and determined whether the routine administration of a trauma screening measure at intake would result in increased diagnoses of posttraumatic stress disorder (PTSD) and in changes in treatment planning in a practice setting. METHODS: A 13-item self-report trauma screening instrument, a shortened version of the Trauma Assessment of Adults instrument, was incorporated into the intake assessment process at a community mental health center (CMHC). A total of 505 out of 515 consumers who presented to the CMHC consecutively were surveyed from May 1, 2001, to January 31, 2002. Data from the initial assessment on trauma exposure and on rate of PTSD diagnosis were examined, and a chart review was conducted on 97 cases (19 percent) to determine the extent to which CMHC services addressed trauma-related problems. RESULTS: Data indicated that 460 consumers (91 percent) had been exposed to one or more traumatic life experiences. The number of traumatic events was negatively correlated with physical and mental health functioning on the 12-item Short-Form Health Survey (SF-12). Subjects with a history of sexual abuse scored significantly higher on the SF-12, reflecting poorer physical and mental health. Although the rate of PTSD diagnosis increased after implementation of the trauma screening instrument, the rates of actual PTSD treatment services provided did not change. CONCLUSIONS: This study strongly suggests that screening for trauma history should be a routine part of mental health assessment and may significantly improve the recognition rate of PTSD. However, much work remains to be done in implementing appropriate treatment. PMID- 14762241 TI - Psychiatric emergencies after the terrorist attacks of September 11, 2001. AB - OBJECTIVE: Population surveys suggest that the events of September 11, 2001, resulted in psychiatric emergencies in U.S. communities. This study tested the extent of such emergencies in San Francisco. METHOD: S: Interrupted time-series designs were applied to counts of emergency calls to the police during the 424 day period beginning January 1, 2001, and of voluntary and coerced admissions to psychiatric emergency services during the 1620-day period beginning July 1, 1997. RESULTS: The number of men and women who were coerced into treatment increased significantly on Thursday, September 13, but the number of voluntary admissions was as expected. The number of telephone calls from citizens that police dispatchers judged to be mental health related increased significantly on Wednesday, September 12, and remained elevated through September 13. Several additional analyses were conducted to test the stability of the findings, and the results were essentially unchanged. CONCLUSIONS: The events of September 11 may not have induced emergent mental illness in U.S. communities at relatively great distance from the attacks. However, it is possible that persons with severe mental illness were either more evident to or less tolerated by the community. PMID- 14762242 TI - Ethnic differences in substance abuse treatment retention, compliance, and outcome from two clinical trials. AB - OBJECTIVE: This study examined the results of two previous studies that evaluated African Americans and whites who were undergoing treatment for cocaine dependence to determine whether the groups differed in pretreatment characteristics, treatment retention, compliance, and cocaine use outcome. METHODS: Data were taken from two trials (N=111 in each), in which patients were randomly assigned to groups that used different behavioral treatments (cognitive-behavioral treatment and 12-step facilitation) and pharmacotherapies (desipramine and disulfiram). RESULTS: Few differences between African Americans and whites were found in terms of demographic characteristics, reasons for seeking treatment, or expectations of treatment. In both studies African Americans and whites did not differ significantly with respect to cocaine use outcomes, but African-American participants completed significantly fewer days of treatment than white participants. In study 2, which was not placebo controlled, African Americans who received disulfiram remained in treatment significantly longer than African Americans who did not receive disulfiram. However, in study 1, in which patients took either desipramine or a placebo, no interactions of ethnicity by medication were found. Among patients who expected improvement to take a month or longer in study 1, African Americans remained in treatment for fewer days than whites. CONCLUSIONS: The behavioral therapies evaluated did not significantly differ in effectiveness for African Americans and whites, suggesting that they are broadly applicable across these ethnic groups. Findings also suggest possible strategies for improving retention of African Americans in treatment. Such strategies might include offering treatment with a medication component and better addressing participants' treatment expectations. PMID- 14762243 TI - Medication adherence among psychotic patients before admission to inpatient treatment. AB - OBJECTIVE: Nonadherence to antipsychotic regimens is considered to be one of the main reasons for hospital readmission. This study assessed the extent of nonadherence in the month before inpatient treatment. METHODS: Ninety-five consecutive patients with an ICD-10 diagnosis of a psychotic disorder (F20) were surveyed about adherence at admission to psychiatric inpatient treatment at a hospital in Linz, Austria. Adherence was assessed on the basis of self-report and by interviewing multiple informants. Episodes of inpatient treatment during the year after discharge were assessed prospectively. RESULTS: Fifty-four patients (57 percent) were rated as partially or fully nonadherent. Compared with patients with good adherence, those who were nonadherent had lower mean scores on the Global Assessment of Functioning (41.4 compared with 47.2), more often received compulsory treatment (52 percent compared with 22 percent), more often had impaired insight into their illness at admission (63 percent compared with 24 percent) and discharge (41 percent compared with 13 percent), and had more days of inpatient treatment in the year after discharge from the index episode (mean of 44.8 days compared with 20.6 days). Adherence was significantly better among patients in regular contact with their psychiatrist and among younger patients. Nonadherent patients who gained insight during treatment had significantly fewer days of inpatient treatment during the next year than those whose insight was still low at discharge (mean of 19.2 days compared with 73.2 days). CONCLUSIONS: Nonadherence is an important contributor to the need for inpatient treatment and is associated with a less favorable course of treatment. The best predictor of further inpatient treatment is insight into illness at discharge. PMID- 14762244 TI - Private health insurance coverage for substance abuse and mental health services, 1995 to 1998. AB - Four years of data from the National Household Survey on Drug Abuse were combined to examine the characteristics of underinsurance in a sample of privately insured Americans aged 18 to 64. Among these adults, 38 percent (45 million) reported not having behavioral health coverage or not knowing their coverage. Young adults aged 18 to 25, Hispanics, Asians, adults in the lowest income level, and less educated adults were more likely to be underinsured. Untreated addictive and psychiatric problems are costly to society. Underinsurance among socially disadvantaged subgroups deserves greater attention from researchers and policy makers. PMID- 14762245 TI - Characteristics of violent behavior in a large state psychiatric hospital. AB - Violent behavior is a significant problem in psychiatric hospitals. The authors reviewed hospital incident reports to identify the characteristics of violent behavior in a large state psychiatric hospital. They found that a very small percentage of patients accounted for a majority of violent episodes, that rates of violent behavior varied among hospital units, that assaultive behavior was more common than self-harm in the long-term units, and that most commonly the assault victims were other patients. The data support earlier studies demonstrating that a small number of patients are responsible for a majority of violent episodes in a hospital setting. PMID- 14762246 TI - Factors associated with stigmatization of persons with mental illness. AB - Stigmatization of individuals with mental illnesses is widespread and serves as a major barrier to treatment. In a survey of 116 undergraduates, the authors examined the impact of diagnosis, attitudes about treatment, and psychiatric terminology on stigma associated with mental illness. Stigmatization of schizophrenia was significantly higher than stigmatization of depression. More positive attitudes toward treatment were associated with significantly less stigma. However, psychiatric terminology had no impact on attitudes toward mental illness. Significantly less stigmatization of mental illness was found among females than among males. Reducing the stigmatization of mental illness continues to be an important goal for mental health professionals. PMID- 14762247 TI - The impact of an employees' strike on a community mental health center. PMID- 14762249 TI - A single funding source for public-sector care. PMID- 14762250 TI - Borderpath for cluster B personality disorder? PMID- 14762251 TI - Referral to aftercare and rehospitalization. PMID- 14762253 TI - Bridging psychiatric services between Asia and America. PMID- 14762254 TI - Unethical to speculate? PMID- 14762255 TI - Studying host immune responses against duck hepatitis B virus infection. PMID- 14762256 TI - Measurement of cell-mediated immune response in woodchucks. PMID- 14762257 TI - Study of liver-specific expression of cytokines during woodchuck hepatitis virus infection. PMID- 14762258 TI - Induction of anti-hepatitis B virus immune responses through DNA immunization. PMID- 14762259 TI - Monitoring gene expression using DNA microarrays during hepatitis B virus infection. PMID- 14762260 TI - Determination of hepatitis B virus-specific CD8+ T-cell activity in the liver. PMID- 14762261 TI - Determining the precursor frequency of HBV nucleocapsid antigen-specific T cells. PMID- 14762262 TI - Detection and characterization of virus-specific CD8+ T cells using the tetramer approach. PMID- 14762263 TI - In vitro analysis of hepatitis B virus specific CD4+ T cells. PMID- 14762264 TI - Induction of humoral and cellular immune responses to hepatitis delta virus through DNA immunization in BALB/c mice. PMID- 14762265 TI - Infection of primary chimpanzee hepatocytes with recombinant hepatitis D virus particles: a surrogate model for hepatitis B virus. PMID- 14762266 TI - Study of the endocytosis and intracellular localization of subviral particles of hepatitis B virus in primary hepatocytes. PMID- 14762267 TI - The tupaia model for the study of hepatitis B virus: direct infection and HBV genome transduction of primary tupaia hepatocytes. PMID- 14762268 TI - Delivery of hepatitis B virus therapeutic agents using asialoglycoprotein receptor-based liver-specific targeting. PMID- 14762269 TI - Woodchuck hepatitis virus hepatocyte culture models. PMID- 14762270 TI - Duck hepatitis B virus primary hepatocyte culture model. PMID- 14762271 TI - Enhancement of infection of HepG2 cells in culture by predigestion of hepadnavirus with V8 protease. PMID- 14762272 TI - Construction of recombinant adenoviruses that produce infectious hepatitis B virus. PMID- 14762273 TI - Baculovirus-mediated gene transfer for the study of hepatitis B virus. PMID- 14762274 TI - Transgenic hepatitis B virus mouse model in the study of chemotherapy. PMID- 14762275 TI - Transplantation of human hepatocytes in immunodeficient UPA mice: a model for the study of hepatitis B virus. PMID- 14762276 TI - Duck hepatitis B virus model in the study of hepatitis B virus. PMID- 14762277 TI - Hepatitis B virus transgenic severe combined immunodeficient mouse model of acute and chronic liver disease. PMID- 14762278 TI - The chimpanzee model: contributions and considerations for studies of hepatitis B virus. PMID- 14762279 TI - Hepatitis B in liver transplant recipients as a special model of antiviral drug development. PMID- 14762280 TI - Hepatitis delta virus transfection for the mouse in vivo model. PMID- 14762281 TI - Hepatitis delta virus RNA transfection for the cell culture model. PMID- 14762282 TI - Analysis of hepatitis B virus dynamics and its impact on antiviral development. PMID- 14762283 TI - Genotyping anti-hepatitis B virus drug resistance. PMID- 14762284 TI - A parsimonious method for screening drug combinations for antihepadnaviral activity using a parametric dose-response surface approach. PMID- 14762285 TI - Hepatitis B virus: where are we and where are we going? PMID- 14762286 TI - Designing studies to evaluate anti-hepatitis B virus therapies: from the perspective of studies for the registration of pharmaceutical products. PMID- 14762287 TI - Specific considerations in the design of hepatitis B virus clinical studies in the far East. PMID- 14762288 TI - Studying the treatment of chronic hepatitis B viral infection in special populations. PMID- 14762289 TI - Designing clinical development programs for anti-hepatitis B virus drugs. PMID- 14762290 TI - Testing antivirals against hepatitis delta virus: farnesyl transferase inhibitors. PMID- 14762291 TI - Detection of HBV DNA in serum using a PCR-based assay. PMID- 14762292 TI - Detection of HBV DNA by oligonucleotide probing. PMID- 14762293 TI - Detection of HBV RNA in serum of patients. PMID- 14762294 TI - Quantification of HBV covalently closed circular DNA from liver tissue by real time PCR. PMID- 14762295 TI - In situ hybridization for the detection and localization of HBV DNA in liver sections. PMID- 14762296 TI - Quantitative assay of hepatitis B surface antigen in serum or plasma using laurell electrophoresis. PMID- 14762297 TI - Detection of hepatitis B virus X antigen by immunohistochemistry and Western blotting. PMID- 14762298 TI - Nonradioisotopic in situ hybridization for HDV RNA. PMID- 14762299 TI - Detection and characterization of small and large HDV antigens. PMID- 14762300 TI - Immunohistochemical detection of hepatitis delta antigen. PMID- 14762301 TI - HBV vaccine-escape variants. PMID- 14762302 TI - HBV genotyping and analysis for unique mutations. PMID- 14762303 TI - A one-filter-three-probe assay for defective interference (DI) effects of naturally occurring core internal deletion (CID) variants of human hepatitis B virus. PMID- 14762304 TI - Detection of hypermodified middle-envelope (M) proteins secreted from naturally occurring HBV variants containing a preS2 internal deletion. PMID- 14762305 TI - Nucleoprotein transport of HBV capsid particles. PMID- 14762306 TI - Phosphorylation analysis of hepatitis B virus core protein in mammalian cells. PMID- 14762307 TI - Study of HBV replication capacity in relation to sequence variation in the precore and core promoter regions. PMID- 14762308 TI - A cis/trans genetic test for pleiotropic phenotypes associated with a frequent naturally occurring mutation at amino acid 97 of HBV core protein. PMID- 14762309 TI - Studying DHBV polymerase by in vitro transcription and translation. PMID- 14762310 TI - Expression and purification of functional hepatitis B virus polymerase in the baculovirus insect cell system. PMID- 14762312 TI - Endogenous polymerase assay for the analysis of hepatitis B virus in transgenic mice. PMID- 14762311 TI - Localization of duck hepatitis B virus polymerase within cells. PMID- 14762313 TI - Transcriptional control of hepatitis B virus. PMID- 14762314 TI - In vitro reconstitution of epsilon-dependent duck hepatitis B virus replication initiation. PMID- 14762315 TI - Hepatitis B viruses : a triple threat for malignant transformation of hepatocytes. PMID- 14762316 TI - How Europe is facing up to antibiotic resistance. PMID- 14762317 TI - Emergence of a new community acquired MRSA strain in Germany. AB - Analysis of community-acquired methicillin-resistant Staphylococcus aureus (c MRSA) from Germany producing the Panton-Valentine leukocidin revealed a unique SmaI-macrorestriction pattern, different from epidemic nosocomial strains. This molecular pattern corresponds to those shown in c-MRSA strains from other countries in the European Union. All isolates exhibited resistance to fusidic acid, which is coded by the far-1 gene. From data on geographical dissemination and time of occurrence, this strain appears to have emerged in Germany in the second half of 2002, and so an already wider dissemination is likely. The emergence of MRSA with resistance to fusidic acid is a first sign of the emergence of a PVL-positive MRSA clone. PMID- 14762318 TI - The European Community strategy against antimicrobial resistance. AB - In 2001 the European Commission presented a 'Community strategy against Antimicrobial Resistance'. In previous years, the problem was addressed through an increasing number of isolated measures, but in this strategy the Commission outlined a comprehensive European Community approach across all sectors. The strategy consists of fifteen actions in four key areas: surveillance, prevention, research and product development, and international cooperation. An important part of this strategy is the 'Council Recommendation on the prudent use of antimicrobial agents in human medicine'. The Recommendation provides a detailed set of public health actions to contain antimicrobial resistance. This paper presents the eleven points of action of the strategy that are directly related to human medicine, and discusses related European Community activities. Under the new public health programme as well as under the research programme of the European Union, antimicrobial resistance is a key priority. PMID- 14762319 TI - Methods for sentinel virological surveillance of influenza in Europe - an 18 country survey. AB - The European Influenza Surveillance Scheme (EISS) is based on an integrated clinical and virological surveillance model. To assess the comparability of virological data, a questionnaire was sent to participants in June 2002 enquiring about specimen collection, laboratory diagnosis of influenza and tests for other respiratory infections. The results showed differences, but also uniformity in virological data collection methods. Similarities were reported for the specimen collection procedures; the type of swab and the transport conditions were comparable. The diagnostic methods were diverse; differences were seen in the (sub)typing methods, with PCR used most often in western countries. The findings will be helpful for the interpretation of virological data collected by sentinel physicians and for the creation of a Community Network of Reference Laboratories for Human Influenza in Europe. Important objectives of the Community Network include the harmonisation of virological methods and the application of quality assurance assessments for the national reference laboratories. PMID- 14762320 TI - Latinos' health care access: financial and cultural barriers. AB - This study aimed at investigating how income, culture, and language affect health care access. Data from a structured questionnaire administered to a random sample of 206 Latinos was analyzed using multivariate logistic regression. Qualitative data served to explain quantitative results. Point estimates for various access measures were similar to national data. In multivariate logistic regression, income and education determined having health insurance (OR 6.8 and 7.4; 95% CI 2.7-17.3 and 2.9-19.0, respectively). Time in the U.S. and health insurance determined having a regular source of care (OR 4.6 and 5.8; 95% CI 1.7-12.8 and 2.1-16.0, respectively). Having a source of care and being female determined visit to the doctor in the past year (OR 6.14 and 6.73; 95% CI 2.3-16.5 and 2.4 19.3, respectively). Language and culture showed no statistically significant effect on access measures, but qualitative data showed they were related to health care barriers. PMID- 14762321 TI - Patterns of health services utilization by recent immigrants. AB - This study was undertaken to analyze how the way young, recently immigrated, families utilize health services evolves over time. Twenty families participated in the study. They all included at least one child of preschool age, had immigrated less than 8 years previously, and had used primary healthcare services since their arrival. A triphasic pattern of utilization was observed, consisting of contact with one or more health services, selection of specific services from those available, and consolidation of choices. Families relied upon a variety of information sources in each of these phases. The primary attributes upon which the families based their evaluation, selection, and adoption of health services were geographical and temporal accessibility, interpersonal and technical quality of services, and language spoken by health professionals and staff. Perception of health services' attributes is influenced by the families' sociocultural referents and preemigration experience. Results indicate that utilization of primary healthcare services progressively changes over time, evolving from the ad hoc use of walk-in services to the adoption of regular sources of care. PMID- 14762323 TI - Cancer incidence patterns among Chinese immigrant populations in Alberta. AB - This study examines the incidence of cancer among Chinese immigrants to Alberta in comparison to the incidence in Canadian-born Alberta residents and in people of Chinese origin still living in China. Cancer cases among Chinese immigrants and Canadian-born Alberta residents were identified from the Alberta Cancer Registry (1974-1993). Incidence rates for Shanghai (1975-1992) were obtained from the International Agency for Research on Cancer (IARC) publications. Direct age standardized incidence rates (ASIRs) were calculated using the "world standard population." Descriptive analysis and Poisson regression modelling were employed to obtain the rate ratios for certain cancer sites among the three populations. For males, the overall incidence of cancer (excluding non-melanoma skin cancer) was lowest in Chinese immigrants while being similar in Canadian-born Alberta residents and Chinese living in Shanghai (197 vs. 224 and 232/100,000). For females, the overall incidence in Chinese immigrants was lower than Canadian-born Alberta residents but similar to that in Chinese living in Shanghai (154 vs 200 and 150/100,000). For cancers that are common in China (stomach and esophagus), the incidence rates for Chinese immigrants were more similar to those for Canadian-born residents than to rates for Shanghai. However, the incidence of liver cancer was very high in the immigrants, suggesting the possible presence of an initiating event during childhood or early adulthood. For cancers that are traditionally uncommon in China (breast and prostate), rates for immigrants were mid-way between those of the two comparison groups. This study supports observations that the risk of cancer in immigrants tends towards the risk of people in the new host country. PMID- 14762324 TI - [Pathophysiology and treatment of intractable sinusitis]. PMID- 14762322 TI - Ethical issues in conducting migrant farmworker studies. AB - Migrant farmworkers should be considered a vulnerable population because they work in a hazardous industry, are often members of an ethnic minority, have known difficulty in accessing health care, and are often of lower socioeconomic status. For these reasons, too, it is extremely important to conduct health-related research with this often-underserved group. However, because migrant farmworkers are vulnerable, investigators must be especially vigilant in protecting them from the potential harms of research and in ensuring that the special ethical issues that arise in research with this population are identified and addressed for every project. In response to the National Cancer Institute's concerns about the feasibility of conducting epidemiologic studies among migrant farmworkers, researchers undertook four feasibility studies near the Texas-Mexico border. Each study raised different, complex ethical questions that challenged the investigators, but whose resolution turned out to be crucial to the success of the studies. PMID- 14762325 TI - [Allergic ocular diseases; from mechanisms through treatments]. PMID- 14762326 TI - [Human antibodies to cedar pollen secreted by human peripheral lymphocytes transformed by Epstein-Barr virus]. AB - To establish immortalized human B-cells secreting antibodies to cedar pollen allergens, peripheral blood lymphocytes from 13 donors were transformed with Epstein-Barr virus. Of 5000 micro culture wells with transformed cell growth, supernatants from 88 wells were found to contain antibodies to pollen allergens. Fourteen supernatants reacted with a cedar allergen Cry j1 and 10 reacted with Cry j2. IgM class antibodies were predominant. PMID- 14762327 TI - [Effect of pranlukast on histamine release and leukotriene c4 (ltc4) generation from human peripheral basophils]. AB - Human basophils may play a role in bronchial asthma. The effect of pranlukast on histamine release and LTC4 generation from human peripheral basophils was examined. Histamine release induced by FMLP and C5a was inhibited by pranlukast in concentration-dependent manner, whereas anti-IgE Ab-induced histamine release was not affected. Both anti-IgE Ab-and FMLP-induced LTC4 were inhibited by pranlukast. These results suggest that pranlukast may improve symptoms of bronchial asthma by inhibiting basophil function in addition to antagonizing Cys LT receptor. PMID- 14762328 TI - [Immediate type food hypersensitivity associated with atopic dermatitis in children]. AB - Food allergy is frequently associated with atopic dermatitis (AD) in children. Appropriate elimination diet is necessary in the case of immediate food hypersensitivity, regardless it causes worsening of the chronic eczema or not. Here we report the prevalence of immediate type food allergy diagnosed by oral food challenge or the episodes of apparent acute allergic reaction in the AD patients (n=182, average age 4.9+/-5.1), who visited our clinic within one year. The prevalence of food allergy in the AD patients was 85.7% in age 0 years, 75.6% in age 1,65.4% in age 2, and declined to 13.9% in age 7 years old or more. The offending foods were egg, milk, wheat, fish and so on. The symptoms of food allergy included skin, gastrointestinal or respiratory manifestations, and also anaphylaxis. In conclusion, immediate type food allergy is frequently associated with childhood AD, and appropriate elimination of the offending food is necessary to avoid the acute allergic reaction including anaphylaxis. PMID- 14762329 TI - [A case of latex allergy suspected drug allergy]. AB - A 30-year-old female with paroxysmal supraventricular tachycardia (PSVT) underwent catheter ablation. About 30 minutes later, urticaria and dyspnea occurred suddenly. Blood pressure decreased to 62/41 mmHg, and she fell into the state of anaphylactic shock. She recovered within one hour following treatment. We initially suspected the onset of anaphylaxis was caused by either the local anesthetic or the intravenous antibiotic administered. Following thorough investigation (skin tests and challenge tests), we concluded that the anaphylaxis was not drug induced. Subsequently, we suspected latex allergy. Skin prick test showed a positive reaction to rubber gloves. The specific test for IgE antibody against latex was positive at 10.8 UA/ml. From these results, anaphylactic shock caused by latex (probably medical gloves) was diagnosed. Doctors should take preventive measures against latex allergy not only in operating rooms but also during minor treatments. It is possible that latex allergy is responsible for some cases of anaphylaxis of unknown origin. PMID- 14762330 TI - [A case of the latex-induced anaphylaxis by contact with barium enema catheter]. AB - The subject was a 34-year-old female with asthma and atopic dermatitis who had previously undergone a Cesarean section. In December 1995, the patient had an acute abdominal pain, so she underwent a barium enema examination. During the procedure, severe anaphylactic shock developed, and the examination was stopped. The patient responded well to appropriate emergency therapy, and her symptoms were resolved. However, the woman visited our outpatient clinic because the symptoms of her atopic dermatitis got worse. She indicated that she felt itchy when using rubber gloves. Also, at the age of thirty, she had urticaria and dyspnea after drinking a glass of fruit juice. Subsequently radioallergosorbent testing demonstrated the presence of specific IgEs against latex, banana, kiwi, grapefruit, and avocado. Skin prick tests were positive for banana, grapefruit, avocado, and latex extract at a dilution of 1:1000. A scratch test was positive to an extract of a balloon catheter which included 0.3 microg/g latex proteins by the LEAP method. In conclusion, the patient was diagnosed with an anaphylaxis to the latex contained in a balloon catheter used for the barium enema. PMID- 14762331 TI - Child passenger deaths involving drinking drivers--United States, 1997-2002. AB - Motor-vehicle crashes are the leading cause of death among children aged >/=1 year in the United States, and one in four crash-related deaths among child passengers aged or = 5% following revascularization was considered clinically significant. RESULTS: Dobutamine stress echocardiography revealed that 489 (37%) of the 1329 dysfunctional segments were viable. A total of 61 (52%) patients had a substantial amount of viable myocardium (> or = 4 viable segments). In these 61 patients the global function was expected to recover > or = 5% after revascularization. However, left ventricular ejection fraction did not improve in 20 (33%) of 61 patients despite the presence of substantial viability. Clinical characteristics and echocardiographic data were comparable between patients with and without improvement. However, patients without improvement had considerably larger end systolic volumes (153 +/- 41 mL vs 133 +/- 46 mL, P =.007). The likelihood of recovery of global function decreased proportionally with the increase of end systolic volume (P <.001, R = 0.43, n = 61). Receiver operating characteristic curve analysis demonstrated that an end systolic volume > or = 140 mL had the highest sensitivity/specificity to predict the absence of global recovery. CONCLUSIONS: In patients with ischemic cardiomyopathy not only the amount of dysfunctional but viable myocardium but also the extent of left ventricular remodeling determines the improvement in function following myocardial revascularization. Patients with a high end systolic volume due to left ventricular remodeling have a decreased likelihood of improvement of global function. PMID- 14762344 TI - Endotracheal calcineurin inhibition ameliorates injury in an experimental model of lung ischemia-reperfusion. AB - OBJECTIVES: We previously demonstrated that calcineurin inhibitors given intravenously ameliorate experimental lung ischemia-reperfusion injury. This study evaluates whether these effects can be achieved when these agents are delivered endotracheally. METHODS: Left lungs of Long Evans rats were rendered ischemic for 90 minutes and reperfused for up to 4 hours. Treated animals received tacrolimus endotracheally at doses of 0.2, 0.1, or 0.025 mg/kg 60 minutes before ischemia. Injury was quantitated in terms of vascular permeability. Additional animals treated at a dose of 0.1 mg/kg were assessed for lung tissue myeloperoxidase content and bronchoalveolar lavage leukocyte content. Bronchoalveolar lavage fluid was assessed for cytokine and chemokine content by enzyme-linked immunosorbent assay. Tissue samples were processed for nuclear factor-kappaB activation, and blood levels of tacrolimus were measured in treated animals. RESULTS: Left lung vascular permeability was reduced in treated animals in a dose-dependent fashion compared with controls. The protective effects correlated with a 47% (0.50% +/- 0.06% vs 0.27% +/- 0.08%, respectively) reduction in tissue myeloperoxidase content (P <.004) and marked reductions in bronchoalveolar lavage leukocyte accumulation. This protection was also associated with decreased nuclear factor-kappaB activation and diminished expression of proinflammatory mediators. Blood tacrolimus levels in treated animals at 4 hours of reperfusion were undetectable. CONCLUSIONS: Tacrolimus administered endotracheally is protective against lung ischemia-reperfusion injury in our model. This protection is associated with a decrease in nuclear factor-kappaB activation. This route of tacrolimus administration broadens its potential clinical use and decreases concerns about systemic and renal toxicity. It may be a useful therapy in lung donors to protect against lung ischemia reperfusion injury. PMID- 14762346 TI - Aortic root surgery in Marfan syndrome: Comparison of aortic valve-sparing reimplantation versus composite grafting. AB - OBJECTIVE: The objective of this study was to compare the results of aortic valve sparing reimplantation and aortic root replacement with mechanical valve conduits in patients with Marfan syndrome undergoing operation for aortic root aneurysms. Patients and methods Between March 1979 and April 2002, 119 patients with clinical evidence of Marfan syndrome underwent composite graft replacement with mechanical valve conduits (n = 74) or aortic valve-sparing reimplantation according to David (n = 45). The underlying causes were aortic dissection type A (43 patients) and aneurysms (76 patients). RESULTS: Patients undergoing aortic valve reimplantation were younger compared with patients undergoing composite grafting (28 vs 35 years, P =.002) and had longer intraoperative aortic crossclamp times (125 vs 78 minutes, P <.0001) and extracorporeal circulation times (162 vs 124 minutes, P <.0001). Early postoperative mortality was 6.8% (n = 5) in patients undergoing composite grafting and 0% in patients undergoing aortic valve reimplantation (P =.15). Mean follow-up was 30 months for patients undergoing aortic valve reimplantation and 114 months for patients undergoing composite grafting. Freedom from reoperation and death after 5 years postoperatively was 92% and 89% in patients undergoing composite grafting and 84% and 96% in patients undergoing aortic valve reimplantation (P =.31; P =.54), respectively. Thromboembolic complications or late postoperative bleeding occurred in 17 patients undergoing composite grafting, and an early postoperative event occurred in 1 patient undergoing aortic valve reimplantation. CONCLUSIONS: The results of aortic valve reimplantation and composite grafting of the aortic valve and ascending aorta with mechanical valve conduits are similar with regard to early and mid-term postoperative mortality and to the incidence of late reoperations in patients with Marfan syndrome. The low risk of thromboembolic or bleeding complications favors aortic valve reimplantation in these patients. PMID- 14762347 TI - Decellularization protocols of porcine heart valves differ importantly in efficiency of cell removal and susceptibility of the matrix to recellularization with human vascular cells. AB - OBJECTIVE: We compared 3 different decellularization protocols in porcine heart valves for efficiency of complete cell removal and potential for recellularization. METHODS: Porcine aortic and pulmonary roots were treated with trypsin, sodium-dodecyl-sulphate, or a new method using 0.25% tert-octylphenyl polyoxyethylen in combination with sodium-deoxycholate. After a subsequent ribonuclease digestion, specimens were seeded with in vitro expanded human saphenous vein endothelial cells and myofibroblasts. RESULTS: After treatment with trypsin and subsequent ribonuclease digestion, endothelial attachment took place; however, xenogenic cells were still visible within the matrix. Unexpectedly, when human cells were seeded onto specimens that had been decellularized with sodium-dodecyl-sulphate, the matrices were surrounded by nonviable endothelial cell fragments, indicating a toxic influence of the ionic detergent; 0.25% tert-octylphenyl-polyoxyethylen together with sodium deoxycholate completely removed porcine cells and enabled host recellularization. CONCLUSION: Compared with trypsin and sodium-dodecyl-sulphate involving decellularization procedures, reported to be effective in cell removal and susceptible to recellularization with human cells, only the porcine matrix treated with a new detergent-based decellularization method using 0.25% tert octylphenyl-polyoxyethylen/sodium-deoxycholate followed by nuclease digestion presented an excellent scaffold for recellularization with human cells. PMID- 14762348 TI - Propensity case-matched analysis of off-pump coronary artery bypass grafting in patients with atheromatous aortic disease. AB - OBJECTIVE: Atheromatous aortic disease is a risk factor for excessive mortality and stroke in patients undergoing coronary artery bypass grafting. Outcomes of off-pump coronary artery bypass grafting and coronary artery bypass grafting with cardiopulmonary bypass in patients with severe atheromatous aortic disease were compared by propensity case-match methods. METHODS: Routine intraoperative transesophageal echocardiography identified 985 patients undergoing isolated coronary artery bypass grafting with severe atheromatous disease in the aortic arch or ascending aorta. Off-pump coronary artery bypass grafting was performed in 281 patients (28.5%). Propensity matched-pairs analysis was used to match patients undergoing off-pump coronary artery bypass grafting (n = 245) with patients undergoing coronary artery bypass grafting with cardiopulmonary bypass. RESULTS: Univariate analysis revealed decreased hospital mortality (16/245, 6.5% vs 28/245, 11.4%; P =.058) and stroke prevalence (4/245, 1.6% vs 14/245, 5.7%; P =.03) in off-pump coronary artery bypass grafting compared with coronary artery bypass grafting with cardiopulmonary bypass. Freedom from any postoperative complication was higher in off-pump coronary artery bypass grafting compared with coronary artery bypass grafting with cardiopulmonary bypass (226/245, 92.2% vs 196/245, 80.0%; P <.001). Multivariable analysis of preoperative risk factors showed that increased hospital mortality was associated with coronary artery bypass grafting with cardiopulmonary bypass (odds ratio = 2.7; P =.01), fewer grafts (P =.05), acute myocardial infarction (odds ratio = 11.5; P <.001), chronic obstructive pulmonary disease (odds ratio = 2.4; P =.03), previous cardiac surgery (odds ratio = 10.2, P =.05), and peripheral vascular disease (odds ratio = 2.1; P =.05). Cardiopulmonary bypass was the only independent risk factor for stroke (odds ratio = 3.6, P =.03). At 36 months' follow-up, comparable survival was observed in the off-pump coronary artery bypass grafting and coronary artery bypass grafting with cardiopulmonary bypass groups (74% vs 72%). Multivariable analysis revealed that renal disease (P <.001), advanced age (P <.001), previous myocardial infarction (P =.03), and lower number of grafts (P =.02) were independent risks for late mortality. CONCLUSIONS: Patients with severe atherosclerotic aortic disease who undergo off-pump coronary artery bypass grafting have a significantly lower prevalence of hospital mortality, perioperative stroke, and overall complications than matched patients who underwent coronary artery bypass grafting with cardiopulmonary bypass. Routine intraoperative transesophageal echocardiography identifies severe atheromatous aortic disease and directs the choice of surgical technique. PMID- 14762349 TI - Increased morbidity and mortality in patients with antiphospholipid syndrome undergoing valve replacement surgery. AB - OBJECTIVE: Heart valve abnormalities are commonly found in patients with antiphospholipid syndrome but experience with valve replacement in such patients is limited. We analyzed the results of valve replacement in patients with this condition at our institution. METHODS: Between 1989 and 2002, 10 patients with antiphospholipid syndrome (8 women, 2 men; aged 38-73 years, mean 49 years) with severe mitral valve disease (n = 7), aortic valve disease (n = 2), or combined mitral-aortic disease (n = 1) underwent valve replacement. We reviewed retrospectively their clinical data, operative and postoperative courses, and the long-term results. Pathological reassessment was performed in all cases. RESULTS: Procedures performed included mitral valve replacement in 7 patients, aortic valve replacement in 2 patients, and combined aortic valve replacement plus mitral valve replacement in 1 patient. In addition, 2 patients underwent tricuspid annuloplasty. The immediate mortality was 20% (2 patients). Major complications occurred in 2 other cases. During a follow-up period of up to 8 years, 2 patients required repeat operation for valve-related complications (1 death). An additional patient died of cardiac causes 13 months after surgery. One patient had major thromboembolic events 3 and 10 months after the operation. The late outcome was uneventful in only 4 patients. CONCLUSION: Valve replacement in patients with antiphospholipid syndrome may carry significant early and late mortality and morbidity, particularly when such patients are referred with advanced valvular heart disease. PMID- 14762350 TI - Clinical features and long-term outcome of type A and type B intramural hematoma of the aorta. AB - OBJECTIVE: Most previous reports on intramural hematoma of the aorta have focused on the initial episode. The purpose of this study was to clarify the long-term outcome of intramural hematoma of the aorta. METHODS: Ninety-four cases of intramural hematoma of the aorta (41 type A and 53 type B) were reviewed. There were 69 male and 25 female patients, and their mean age was 66.7 +/- 8.7 years (range, 46-88 years). RESULTS: Eleven (27%) of the patients with type A hematoma and 1 (2%) of the patients with type B hematoma underwent early surgical intervention. Others were treated medically, and the overall hospital mortality was 7% for patients with type versus 2% for patients with type B intramural hematomas of the aorta (P =.315). Twenty-three patients, 9 (22%) with type A and 14 (26%) with type B intramural hematomas of the aorta, underwent late surgical intervention during the follow-up period, and there were no hospital deaths. A total of 23 patients died during the follow-up period, including 6 of intramural hematoma of the aorta-related deaths (3 in the type A group and 3 in the type B group). The estimated freedom from intramural hematoma of the aorta-related events at 1 and 5 years was 70% +/- 8% and 54% +/- 11% for the type A group versus 73% +/- 6% and 58% +/- 8% for the type B group, respectively (P =.972). After excluding the nonintramural hematoma of the aorta-related deaths, the survival rates at 5 and 10 years were 80% +/- 9% and 80% +/- 9% for the type A group and 91% +/- 8% and 81% +/- 11% for the type B group (P =.211). CONCLUSIONS: Intramural hematoma of the aorta-related events occur equally in both types of intramural hematoma of the aorta. We recommend close follow-up for at least 5 years because most intramural hematoma of the aorta-related events occur during this period. PMID- 14762351 TI - Coronary artery bypass with ventricular restoration is superior to coronary artery bypass alone in patients with ischemic cardiomyopathy. AB - BACKGROUND: Coronary artery bypass is an acceptable therapy in patients with ischemic cardiomyopathy. However, it has been demonstrated that patients with increased left ventricular volume have a worse outcome than patients with normal ventricular volume. Our hypothesis was that ventricular restoration plus coronary artery bypass provides improved outcome compared with coronary artery bypass alone in ischemic cardiomyopathy with ventricular enlargement. METHODS: A retrospective analysis was performed of patients with ischemic cardiomyopathy (ejection fraction <30%) who underwent operation between 1998 and 2002. Patients with enlarged ventricles (end-diastolic dimension > or =6.0 cm) who underwent either coronary artery bypass alone or coronary artery bypass with ventricular restoration were compared. Preoperative and postoperative ejection fraction, morbidity, mortality, and freedom from heart failure (hospitalization secondary to heart failure) were assessed. RESULTS: Ninety-five patients were included in the study. Thirty-nine patients had coronary artery bypass alone, whereas 56 patients had ventricular restoration with coronary artery bypass. Both groups demonstrated an improved postoperative ejection fraction; however, the improvement was significantly greater in the ventricular restoration plus coronary artery bypass group (P <.01). There were no hospital deaths in either group; however, late mortality was higher in the coronary artery bypass group. Freedom from heart failure was achieved in all but 2 of the ventricular restoration plus coronary artery bypass patients (2/56, or 3.6%) versus 7 in the coronary artery bypass group (7/39, or 18%). The combined outcomes of freedom from failure and late mortality were significantly improved in the ventricular restoration plus coronary artery bypass group (P <.05). CONCLUSIONS: Ventricular restoration affords significant improvement in ejection fraction compared with coronary artery bypass alone, without added mortality. Most importantly, left ventricular restoration reduces late morbidity and mortality compared with coronary artery bypass alone in patients with large ventricles. PMID- 14762352 TI - Effect of surgical revascularization of a right coronary artery tributary of an infarcted nonischemic territory on the outcome of patients with three-vessel disease: a prospective randomized trial. AB - BACKGROUND: We evaluated the in-hospital and long-term effects of surgical grafting of a dominant graftable right coronary artery tributary of an infarcted nonischemic territory in patients with triple-vessel disease who were undergoing coronary artery bypass grafting. METHODS: Of 303 consecutive patients undergoing coronary artery bypass grafting with 3-vessel coronary disease and a dominant right coronary artery tributary of an infarcted nonischemic territory, 154 were randomized to right coronary artery revascularization and 149 to no right coronary artery grafting. In all cases, standard on-pump surgical myocardial revascularization was performed. RESULTS: Overall hospital mortality was 2 of 154 versus 1 of 149 (P =.97); no difference in in-hospital outcome was observed between the 2 groups. At follow-up, cardiac event-free survival was 84 of 152 in the right coronary artery grafting series and 62 of 148 in the non-right coronary artery grafting group (P =.20). However, when the analysis was limited to surviving patients without new scintigraphic evidence of ischemia (to avoid confounding factors derived from ischemia in the left coronary system or right coronary artery graft malfunction), we found that patients who received a right coronary artery graft had fewer cardiac events, a lower incidence of arrhythmia, and less left ventricular dilatation than did the non-right coronary artery revascularized series. CONCLUSIONS: Surgical grafting of a right coronary artery tributary of an infarcted nonischemic territory in patients with 3-vessel coronary artery disease submitted to coronary artery bypass grafting improved late electric stability, ventricular geometry, and event-free survival but did not affect in-hospital or 10-year survival. PMID- 14762353 TI - Neochordal repair of the posterior mitral leaflet. AB - BACKGROUND: Myxomatous mitral valve insufficiency is traditionally repaired by posterior leaflet quadrangular resection and reconstruction. A simplified repair technique without leaflet resection is described, and our initial experience is reviewed. METHODS: Thirty-nine consecutive patients with significant mitral regurgitation underwent repair since January 2000 by placement of expanded polytetrafluoroethylene sutures between the leading (coapting) edge of the posterior leaflet and the corresponding papillary muscle. An annuloplasty ring was placed, and no leaflet tissue was resected. Patient medical records were obtained and retrospectively reviewed. RESULTS: Twenty-five men and 14 women (median age, 61 years; range, 40-88 years) had their mitral valve repaired by a variety of surgical approaches, including robotic (18 patients), right thoracotomy (6 patients), and sternal (15 patients). Three patients have required valve replacement: 1 at the initial operation, 1 because of dehiscence of the annuloplasty ring, and 1 after subsequent rupture of a previously normal native chorda. At follow-up (median, 12 months), 92% (33/36) of the remaining patients had an intact mitral repair with no to mild regurgitation, 8.3% (3/36) of patients had moderate regurgitation, and 92% of all patients (36/39) were in New York Heart Association class I. There were no deaths. CONCLUSIONS: Myxomatous mitral regurgitation due to posterior leaflet insufficiency can be repaired without leaflet resection by placement of neochordae. This repair technique is effective and is readily accomplished by traditional and minimally invasive surgical approaches. PMID- 14762354 TI - Hypoplasia of the small pulmonary arteries in total anomalous pulmonary venous connection with obstructed pulmonary venous drainage. AB - OBJECTIVE: Preoperative pulmonary venous obstruction has been reported to be a risk factor negatively impacting survival in total anomalous pulmonary venous connection. We examined lung tissue from total anomalous pulmonary venous connection patients with pulmonary venous obstruction and demonstrated hypoplasia of small pulmonary arteries to elucidate the mechanism underlying the poor outcome. METHODS: Ten total anomalous pulmonary venous connection patients with preoperative pulmonary venous obstruction between the ages of 2 days and 10 months were studied. As histological parameters, we assessed the size of small pulmonary arteries in relation to the size of accompanying bronchioles to identify small pulmonary artery underdevelopment. Other parameters, such as the radial alveolar count, which reflects alveolar maturity, intimal lesions, lymphangiectasia, and the medial thickness of small pulmonary arteries and small pulmonary veins, were also examined. As a control group, we examined 24 autopsy cases with no congenital heart or pulmonary disease. RESULTS: When the radius of the accompanying bronchiole was 100 microm, the radius of small pulmonary artery in the control group was found to enlarge for the first 2 months and then remain stable at approximately 80 microm from 2 to 10 months. In total anomalous pulmonary venous connection with preoperative pulmonary venous obstruction, the radius was significantly lower than in the control (47.0 +/- 21.8 microm versus 75.9 +/- 9.8 microm, P <.001), and the difference between dead and surviving patients was significant at P <.001 (33.0 +/- 14.6 microm versus 68.2 +/- 9.2 microm). Examination of the alveoli yielded an radial alveolar count of 4.6 +/- 1.5 in the control group and 4.4 +/- 0.8 in the total anomalous pulmonary venous connection patients, and the difference was not significant (P =.71). CONCLUSIONS: The small pulmonary arteries of total anomalous pulmonary venous connection patients with preoperative pulmonary venous obstruction were underdeveloped compared with controls but their alveolae were not hypoplastic. These results suggested that the small pulmonary artery hypoplasia may be responsible for the poor outcome of these patients. PMID- 14762355 TI - Occurrence and management of atrial arrhythmia after long-term Fontan circulation. AB - OBJECTIVES: In patients after the Fontan operation, we determined risk factors for late failure and for intra-atrial re-entrant tachycardia at 15 to 20 years' follow-up. Midterm results after electrophysiologic ablation therapy for these tachycardias were also evaluated. METHODS: Current follow-up was available in 162 patients (2005 patient-years) with a wide range of underlying diagnoses operated on between February 1978 and May 1995. Risk factor analysis included patient related and procedure-related variables, with late failure and the incidence of re-entrant tachycardia as outcome parameters. RESULTS: Forty late failures were observed (2.0 per 100 patient-years). At 15 years, Kaplan-Meier estimated survival was significantly (P =.007) better for patients with tricuspid atresia (93%) compared with that for patients with complex congenital malformation (71%). The sole multivariable risk factor for Fontan failure was the type of underlying diagnosis. At 20 years' follow-up, overall freedom from tachycardia was estimated to be 46% +/- 12%. Acute success of electrophysiologic ablation was seen in 25 (83%) of 30 patients, and Kaplan-Meier estimated freedom from recurrent tachycardia was 81% +/- 10% at 3 years. Multivariate analysis identified duration of Fontan circulation as the sole risk factor for re-entrant tachycardias. CONCLUSION: After the modified Fontan operation, long-term survival in patients with tricuspid atresia was significantly better compared with that in patients with complex congenital malformations. As first-choice therapy for atrial re entrant tachycardias, we recommend electrophysiologic ablation therapy. PMID- 14762356 TI - Tricuspid valve repair in hypoplastic left heart syndrome. AB - OBJECTIVES: Currently, the survival for the Norwood procedure for hypoplastic left heart syndrome is approximately 90% in selected centers. However, the development of tricuspid regurgitation remains a significant obstacle to successful staged repair in a subset of these patients. The results of tricuspid valve repair in this challenging patient population remain largely unknown. METHODS: Twenty-eight patients with significant (3-4+) tricuspid regurgitation after the Norwood procedure required tricuspid valve repair from August 1995 through December 2002. The clinical and Doppler-echocardiographic data were reviewed to determine the efficacy of repair and patient outcome. RESULTS: Follow up was 96% complete (27/28). Patients were divided into 2 groups on the basis of tricuspid regurgitation at late follow-up: those with a successful late outcome (0-2+) and those with a poor outcome (3-4+). There were 17 (63%) patients with a successful result and 10 (37%) with an adverse outcome. Age, weight, follow-up duration, valve anatomy, and stage of palliation were not significantly different between groups. Early postoperative 0 to 2+ regurgitation was associated with a durable result (P =.012) and preserved ventricular function (P =.04). Need for repair other than a partial annuloplasty was predictive of a poor outcome (P =.04). Overall survival was 67% (18/27). Survival was 94% (16/17) for patients with a successful late result versus 20% (2/10) for those with a poor outcome (P =.0002). CONCLUSIONS: Tricuspid valve repair can be accomplished in this challenging patient population with excellent results. Successful tricuspid valve repair is predictive of continued good valve function and preserved right ventricular function. Successful valve repair at late follow-up predicts excellent late survival. PMID- 14762357 TI - Redefining the impact of oxygen and hyperventilation after the Norwood procedure. AB - OBJECTIVE: Postoperative management after the Norwood procedure is aimed at optimizing systemic oxygen delivery and mixed venous oxygen saturation. High levels of fraction of inspired oxygen and hyperventilation may increase pulmonary blood flow at the expense of systemic flow. This study determines the effects of these interventions on mixed venous saturation and systemic oxygen delivery in postoperative neonates. METHODS: We prospectively studied the effects of 100% fraction of inspired oxygen and hyperventilation in 14 neonates (median age 8 days) 1 to 3 days after the Norwood procedure, while they were sedated, paralyzed, and mechanically ventilated. After establishment of baseline conditions (fraction of inspired oxygen = 29% +/- 2%, normal ventilation), patients were exposed to each of the 2 interventions in random order. Mixed venous saturation was measured through a transthoracic line in the superior vena cava. Oxygen excess factor (Omega = systemic oxygen delivery/oxygen consumption) was used as an indicator of systemic oxygen delivery. RESULTS: High levels of fraction of inspired oxygen produced significant increases from baseline in systemic saturation (90% +/- 1% vs 80% +/- 1%, P <.01), mixed venous saturation (54% +/- 3% vs 44% +/- 2%, P <.01), and oxygen excess factor (2.6% +/- 0.2% vs 2.3 +/- 0.2%, P <.01), but there was no change in arteriovenous saturation difference or blood pressure. Hyperventilation resulted in no changes in systemic or mixed venous saturation, arteriovenous saturation difference, oxygen excess factor, or blood pressure. CONCLUSIONS: High levels of fraction of inspired oxygen can improve mixed venous oxygen saturation and systemic oxygen delivery after the Norwood procedure. Hyperventilation does not change either mixed venous saturation or oxygen delivery. Management protocols aimed at minimizing the fraction of inspired oxygen and carefully controlling ventilation may not be warranted. PMID- 14762358 TI - Extended septal myectomy for hypertrophic obstructive cardiomyopathy with anomalous mitral papillary muscles or chordae. AB - OBJECTIVES: Transaortic left ventricular septal myectomy yields excellent results for most severely symptomatic patients with hypertrophic obstructive cardiomyopathy. However, associated anomalies of the mitral subvalvular apparatus may prevent complete relief of obstruction, and mitral valve replacement has been advocated. We reviewed our results of procedures designed to relieve obstruction with preservation of the mitral valve. METHODS: Among 291 patients undergoing septal myectomy from 1975 to 2002, 56 (ages 2-77 years) had anomalous mitral subvalvular apparatus including anomalous chordae (n = 28) and papillary muscles with direct insertion into mitral leaflets (n = 13) or fusion to septum (n = 31) or free wall (n = 12); 82% of patients were in New York Heart Association class III or IV. Operation included resection of anomalous chordae (28 patients), relief of papillary muscle fusion (36 patients), and extended septal myectomy, wider at the apex than the base. RESULTS: There were no early deaths and no patients required mitral valve replacement. Mean peak pressure gradients decreased from 70 +/- 28 to 4.9 +/- 8.4 mm Hg and mean mitral regurgitation grade decreased from 2.3 to 1.0 (P <.001). Mean follow-up was 2.8 +/- 2.6 years. Freedom from reoperation at 4 years was 95%. There were 3 late noncardiac deaths; 98% of patients were in New York Heart Association class I or II. CONCLUSIONS: Hypertrophic obstructive cardiomyopathy associated with anomalous mitral papillary muscles or chordae can be successfully treated without mitral valve replacement by surgical relief of the anomalies and an extended septal myectomy; early mortality is low, obstruction and mitral regurgitation are significantly reduced, and late results are excellent. PMID- 14762359 TI - Neonatal vulnerability to ischemia and reperfusion: Cardioplegic arrest causes greater myocardial apoptosis in neonatal lambs than in mature lambs. AB - OBJECTIVES: Apoptosis is a mechanism for deletion of injured or obsolete cells that is distinct from necrosis and mediated by mitochondrial release of cytochrome c caspase activation. Because myocardial apoptosis is a part of normal fetal and postnatal maturation, we hypothesize that neonatal myocardium is more vulnerable to undergo myocardial apoptosis than mature myocardium after cardioplegic arrest. METHODS: Newborn and mature lambs (n = 5 in each group) underwent cardiopulmonary bypass, antegrade crystalloid hyperkalemic cardioplegic arrest for 60 minutes, and a 6-hour recovery period. Myocardium was examined by using terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate digoxigenin nick end labeling (TUNEL), Western blotting, in vitro kinase assays, and fluorometric assays of the activity of caspases 3, 8, and 9. Myocardium from nonoperated control subjects (n = 5 in each age group) was also obtained. RESULTS: More TUNEL-positive nuclei were present in the newborn postcardioplegic myocardium (P =.04). Caspase 3, 8, and 9 activities were 1.6-fold, 1.5-fold, and 1.4-fold greater in the newborn postcardioplegic myocardium (P =.04, P =.01, and P =.01, respectively). The Bax/Bcl-2 ratio was higher in the newborn postcardioplegic myocardium (P =.04). Apoptosis signal-regulating kinase 1 activity and cleaved caspase 3 levels were higher in the newborn postcardioplegic myocardium (P =.02 and P =.009). Mitochondrial release of cytochrome c was greater in the newborn postcardioplegic myocardium (P =.009). CONCLUSIONS: The increased Bax/Bcl-2 ratio in the newborn myocardium suggests a proapoptotic state that is manifested by greater TUNEL staining, cytochrome c release, and cleavage of caspase 3. Increased apoptosis signal-regulating kinase 1 activity suggests greater oxidative stress, immature mechanisms to ameliorate oxidative stress, or both in the neonatal myocardium. Mitochondrial release of cytochrome c suggests that apoptosis-related mitochondrial dysfunction might contribute to early postoperative myocardial dysfunction in the neonate. PMID- 14762360 TI - Stapled coronary anastomosis with minimal intraluminal artifact: The S2 Anastomotic System in the off-pump porcine model. AB - OBJECTIVE: A reliable, easy-to-use, 1-shot anastomotic device will significantly push the barrier for less invasive coronary bypass surgery. The current study was designed to test the safety, efficacy, and early patency of a novel distal anastomotic device. METHODS: The S2 Anastomotic System (iiTech BV, Amsterdam, The Netherlands) was used in 10 consecutive pigs (73 kg) on a mild antiplatelet regimen. In each animal, the device was used to create an internal thoracic artery to left anterior descending bypass on the beating heart. The anastomoses were evaluated intraoperatively (n = 10), at 2 days (n = 2), and at 5 weeks (n = 8) by functional flow measurements, postmortem angiography, and histomorphologic examination. RESULTS: In all pigs, the S2 Anastomotic System rapidly created successful anastomoses at the first attempt (graft loading and coronary ischemia time: 1.2 +/- 0.3 minutes and 3.0 +/- 0.6 minutes) on target vessels of 1.6 to 2 mm inner diameter. There were no technical failures or anastomotic leaks requiring additional sutures. Both intraoperatively and at the time of death, ischemically induced peak hyperemic flow responses demonstrated widely patent bypasses, which were confirmed by postmortem angiography (FitzGibbon grade A, n = 10) and macroscopic evaluation (anastomotic orifice: 2 mm). Histomorphologic evaluation showed a normal healing response with negligible neointima covering the connector and limited streamlining repair tissue formation between the staple like elements of the connector. CONCLUSIONS: The S2 Anastomotic System consistently created automated, fast, and reliable internal thoracic to coronary artery anastomoses on the porcine beating heart with excellent graft patency and healing characteristics at the 5-week follow-up. PMID- 14762361 TI - Robotic totally endoscopic coronary artery bypass: program development and learning curve issues. AB - BACKGROUND: The introduction of new procedures in heart surgery is a critical phase that includes learning curves and the risk of increased mortality or morbidity. Totally endoscopic coronary artery bypass grafting using robotic techniques represents such an innovative procedure. The aim of this report is to demonstrate the safe introduction of totally endoscopic coronary artery bypass grafting using a stepwise and modular approach. METHODS: From June 2001 until December 2002, 50 procedures were performed using the da Vinci telemanipulator system. After baseline training the following procedure modules were carried out in a stepwise manner: robotically assisted endoscopic left internal thoracic artery harvesting and completion of the procedure as conventional coronary artery bypass grafting, minimally invasive direct coronary artery bypass, or off-pump coronary artery bypass (n = 19), robotically assisted suturing of left internal thoracic artery to left anterior descending anastomoses during conventional coronary artery bypass grafting (n = 15), totally endoscopic coronary artery bypass grafting on the arrested heart using remote access perfusion and aortic endocclusion coronary bypass grafting (n = 15). One patient was excluded intraoperatively from a robotic procedure due to pleural adhesions. RESULTS: A significant learning curve was observed for left internal thoracic artery takedown time, y(min) = 181 - 39 x ln(x) (x = procedure number) (P <.001), and total operative time in totally endoscopic coronary artery bypass grafting, y(min) = 595 - 87 x ln(x) x = (procedure number) (P =.028). The conversion rate in totally endoscopic coronary artery bypass grafting was 2/15. Intensive care unit stay correlated significantly with total operative time (r =.427, P =.002). There was no hospital mortality. CONCLUSION: Totally endoscopic coronary artery bypass grafting can be safely implemented into a heart surgery program. Learning curves are steep for robotic left internal thoracic artery takedown and for performance of totally endoscopic coronary artery bypass grafting. Long operative times translate into prolonged intensive care unit stay in specific cases but not into increased mortality. PMID- 14762362 TI - Ventral cardiac denervation reduces the incidence of atrial fibrillation after coronary artery bypass grafting. AB - OBJECTIVES: Because the autonomic nervous system is an important determinant in the appearance of atrial fibrillation, we have assessed the role of ventral cardiac denervation for its prevention. METHODS: Patients undergoing low-risk coronary artery surgery were enrolled. No routine antiarrhythmic drugs were administered before or after the operation. Ventral cardiac denervation was performed in 207 patients, and 219 patients were used as control subjects. Denervation was performed before cardiopulmonary bypass. The groups were comparable regarding demographic, clinical, and operative variables. RESULTS: The additional time for the denervation was 5 +/- 2 minutes, and there were no associated complications. Postoperative atrial fibrillation was present in 15 (7%) patients undergoing ventral cardiac denervation (95% confidence interval, 4% 12%) and in 56 (27%) control subjects (95% confidence interval, 18%-35%). Patients submitted to ventral cardiac denervation had fewer and less severe episodes of atrial fibrillation, and no patient had atrial fibrillation after discharge. Ventral cardiac denervation was the most significant predictor of postoperative atrial fibrillation (odds ratio, 0.42; confidence interval, 0.23 0.78; P =.006). Age of greater than 65 years (odds ratio, 1.67; confidence interval, 0.96-2.9; P =.067) was a highly suggestive predictor. The analysis of the effect of ventral cardiac denervation correlated with the patient's age showed a more pronounced effect in patients younger than 70 years (odds ratio, 0.43; confidence interval, 0.22-0.86; P =.022) CONCLUSIONS: Ventral cardiac denervation is a fast and low-risk procedure. Its use significantly reduces the incidence and severity of atrial fibrillation after routine coronary artery bypass surgery. Patients younger than 70 years of age are expected to have a higher success rate than those older than 70 years. PMID- 14762363 TI - Cardiac denervation after clinical transmyocardial laser revascularization: short term and long-term iodine 123-labeled meta-iodobenzylguanide scintigraphic evidence. AB - OBJECTIVES: This study was designed to investigate whether transmyocardial laser revascularization induces myocardial denervation and to correlate this with myocardial perfusion and clinical status. METHODS: Transmyocardial laser revascularization was performed with a Holmium:YAG (n = 3) or xenon chloride excimer laser (n = 5). Preoperative and postoperative iodine 123-labeled meta iodobenzylguanide SPECT scintigraphy to assess cardiac innervation and perfusion scintigraphy were also performed. Furthermore, New York Heart Association functional angina class and quality of life were assessed. RESULTS: In all patients postoperative iodine 123-labeled meta-iodobenzylguanide SPECT showed significantly decreased uptake and therefore sympathetic myocardial denervation at up to 16 months' follow-up (average preoperative and postoperative summed defect scores of 14.8 +/- 5.3 and 24.5 +/- 4.2, respectively; P =.00005). In 86% of segments, the decreased meta-iodobenzylguanide uptake could be correlated to the treated area. In all patients angina was reduced by 2 or more classes at 12 months' follow-up, and quality of life improved significantly. CONCLUSIONS: Transmyocardial laser revascularization-induced improvement of angina and quality of life can be explained by destruction of nociceptors or cardiac neural pathways, changing the perception of anginal pain. PMID- 14762364 TI - Can the use of methylprednisolone, vitamin C, or alpha-trinositol prevent cold induced fluid extravasation during cardiopulmonary bypass in piglets? AB - OBJECTIVE: Hypothermic cardiopulmonary bypass is associated with capillary fluid leakage, resulting in edema and occasionally organ dysfunction. Systemic inflammatory activation is considered responsible. In some studies methylprednisolone has reduced the weight gain during cardiopulmonary bypass. Vitamin C and alpha-trinositol have been demonstrated to reduce the microvascular fluid and protein leakage in thermal injuries. We therefore tested these three agents for the reduction of cold-induced fluid extravasation during cardiopulmonary bypass. METHODS: A total of 28 piglets were randomly assigned to four groups of 7 each: control group, high-dose vitamin C group, methylprednisolone group, and alpha-trinositol-group. After 1 hour of normothermic cardiopulmonary bypass, hypothermic cardiopulmonary bypass was initiated in all animals and continued to 90 minutes. The fluid level in the extracorporeal circuit reservoir was kept constant at the 400-mL level and used as a fluid gauge. Fluid needs, plasma volume, changes in colloid osmotic pressure in plasma and interstitial fluid, hematocrit, and total water contents in different tissues were recorded, and the protein masses and the fluid extravasation rate were calculated. RESULTS: Hemodilution was about 25% after start of normothermic cardiopulmonary bypass. Cooling did not cause any further changes in hemodilution. During steady-state normothermic cardiopulmonary bypass, the fluid need in all groups was about 0.10 mL/(kg.min), with a 9-fold increase during the first 30 minutes of cooling (P <.001). This increased fluid need was due mainly to increased fluid extravasation from the intravascular to the interstitial space at a mean rate of 0.6 mL/(kg.min) (range 0.5-0.7 mL/[kg.min]; P <.01) and was reflected by increased total water content in most tissues in all groups. The albumin and protein masses remained constant in all groups throughout the study. CONCLUSION: Pretreatment with methylprednisolone, vitamin C, or alpha trinositol was unable to prevent the increased fluid extravasation rate during hypothermic cardiopulmonary bypass. These findings, together with the stability of the protein masses throughout the study, support the presence of a noninflammatory mechanism behind the cold-induced fluid leakage seen during cardiopulmonary bypass. PMID- 14762365 TI - Adenoviral-mediated transfer of vascular endothelial growth factor 121 cDNA enhances myocardial perfusion and exercise performance in the nonischemic state. AB - BACKGROUND: Angiogenic gene therapy has been demonstrated to enhance perfusion to ischemic tissues, but it is unknown whether the administration of angiogenic growth factors will increase blood flow to nonischemic tissues. This study investigates whether enhanced myocardial perfusion can be mediated by adenovirus mediated transfer of vascular endothelial growth factor 121 cDNA to nonischemic myocardium. METHODS: New Zealand White rabbits received adenovirus (5 x 10(10) particle units) encoding for vascular endothelial growth factor 121 (n = 14) or a control vector without a transgene (n = 13) or saline solution (n = 9) via direct myocardial injection. Fluorescent microsphere perfusion studies and histologic analyses were performed 4 weeks later. In a parallel study, exercise treadmill testing was performed to assess the functional effects of this therapy in Sprague Dawley rats. RESULTS: Microsphere assessment of myocardial perfusion in rabbits 4 weeks after adenovirus-encoding vascular endothelial growth factor administration was greater than that for rats injected with control vector without a transgene or saline solution (3.2 +/- 0.5 vs 2.7 +/- 0.7 and 2.4 +/- 0.4, respectively; P <.03). The endothelial cell count per high power field was increased in animals injected with adenovirus-encoding vascular endothelial growth factor versus animals injected with control vector without a transgene or saline solution (147 +/- 27 vs 123 +/- 14 and 125 +/- 16 cells, respectively), although this did not reach statistical significance (P =.12). Rats treated with adenovirus-encoding vascular endothelial growth factor also demonstrated prolonged exercise tolerance compared with rats injected with control vector without a transgene or saline solution (exhaustion time: 26 +/- 5 minutes vs 19 +/- 2 minutes and 20 +/- 3 minutes, respectively; P =.006). CONCLUSIONS: Adenovirus encoding-mediated transfer of vascular endothelial growth factor 121 induces an enhancement in regional perfusion in nonischemic myocardium that corresponds to changes in exercise tolerance. Adenovirus-encoding vascular endothelial growth factor therapy may be useful for inducing angiogenesis in the nonischemic state, such as for prophylactic therapy of early coronary artery disease. PMID- 14762366 TI - Tumor necrosis factor-alpha from resident lung cells is a key initiating factor in pulmonary ischemia-reperfusion injury. AB - OBJECTIVE: A central role of macrophages in initiating lung ischemia-reperfusion injury is emerging. Tumor necrosis factor-alpha is a proinflammatory cytokine secreted mainly by macrophages under various conditions. We hypothesized that tumor necrosis factor-alpha from resident lung cells is a key initiating factor in pulmonary ischemia-reperfusion injury. METHODS: We used an isolated, buffer perfused lung system to explore the role of tumor necrosis factor-alpha production by resident lung cells in pulmonary ischemia-reperfusion injury. Lungs from wild-type mice and tumor necrosis factor-alpha-deficient mice were subjected to 60 minutes of ischemia followed by 60 minutes of reperfusion. Histologic injury scores and measurements of lung compliance, airway resistance, mean pulmonary artery pressure, vascular reactivity, and wet lung weight index were obtained and compared using repeated-measures analysis of variance. RESULTS: Lungs from tumor necrosis factor-alpha-deficient mice showed significantly less injury in all physiologic parameters throughout the entire 60 minutes of reperfusion compared with lungs from wild-type mice (P <.001). The most notable effects were observed in pulmonary artery pressure and airway resistance. Vascular reactivity (acute vasoconstrictive episodes per 60 minutes) was also blunted in the lungs from tumor necrosis factor-alpha-deficient mice compared with the lungs from wild-type mice (5.8 responses/hour vs 1.2 responses). Histologic injury scores and wet lung weight index were significantly reduced in lungs from tumor necrosis factor-alpha-deficient mice. CONCLUSIONS: By using the advantages of a nonblood-perfused system, we have focused our investigation on resident lung cells. Our results demonstrate that resident cell-produced tumor necrosis factor-alpha is a key initiating factor in acute lung ischemia reperfusion injury. PMID- 14762367 TI - Preoperative detection and management of immune heparin-induced thrombocytopenia in patients undergoing heart surgery with iloprost. AB - OBJECTIVE: The objective of this study was to evaluate our protocol for the identification and management of patients with immune heparin-induced thrombocytopenia undergoing cardiac surgery. METHODS: Among 1518 patients who underwent cardiac surgery between June 1998 and May 2001, 32 (2.1%) presented with platelet counts less than 150,000/mm3 preoperatively or a history of prolonged (>3 days) intravenous exposure to heparin or both. These 32 patients were evaluated with an enzyme-linked immunosorbent assay for antibodies against heparin-platelet factor 4 complex. Platelets of patients with detected antibodies were tested with the prostacyclin analog iloprost for inhibition of heparin aggregation and determination of the inhibiting concentration and corresponding intravenous infusion rate of iloprost. Patients with antibodies received heparin after complete platelet inhibition with iloprost infusion. Hypotension was prevented or treated with intravenous noradrenaline. Ten randomly selected patients with similar preoperative characteristics, no previous extended exposure to heparin, and normal platelet counts served as controls. RESULTS: Ten of the 32 patients (group A, 31.3%) and none of the controls had antibodies against heparin platelet factor 4 complex. Patients in group A underwent surgery with iloprost (6 24 ng.kg(-1).min(-1)) and had their blood pressure maintained at greater than 95 mm Hg with norepinephrine infusion (1-4 microg.kg(-1).min(-1)). Operative mortality was zero. There were no thrombotic complications or bleeding requiring exploration. One patient in group A bled 1310 mL/6 hours but did not need exploration. There was no difference in postoperative blood loss and morbidity between groups. Platelet counts were reduced by 12.5% +/- 8.7% (group A) and 38.1% +/- 15.2% (control) (P <.001) 1 hour postoperatively and reached preoperative values by the fifth postoperative day. CONCLUSIONS: Immune heparin induced thrombocytopenia can be detected preoperatively among patients with a low platelet count or a history of prolonged heparin exposure or both. Cardiac surgery can be safely undertaken using iloprost-induced platelet inhibition during heparinization. PMID- 14762369 TI - The technique of unilateral double lobar lung transplantation in a canine model. AB - OBJECTIVE: Bilateral living-donor lobar lung transplantation has become an accepted alternative to cadaveric lung transplantation. Because only one lobe is implanted in each chest cavity, this procedure seems to be best suited for children and small adults. The purpose of this study was to develop a technique of unilateral double lobar lung transplantation that can be applied to large adult patients. METHODS: Unilateral double lobar lung transplantation was performed in 6 weight-matched pairs of dogs. In donor animals the right middle, lower, and cardiac lobes were separated as a right graft, and the left lower lobe was separated as a left graft. In recipient animals these 2 grafts were implanted in the right hemithorax after right pneumonectomy. The left graft was implanted as a right upper lobe, having been rotated 180 degrees along the vertical axis and then 180 degrees along the horizontal axis. The right graft was implanted in the natural anatomic position. Function of the transplanted grafts was assessed for 3 hours after ligation of the left main pulmonary artery while the animals were ventilated with 100% oxygen. RESULTS: Morphologic adaptation of the 2 grafts in the right hemithorax was found to be excellent. All 6 animals survived the assessment period with excellent pulmonary function. At the end of the 3-hour assessment period, the arterial oxygen tension was 519 +/- 31 mm Hg, and the mean pulmonary artery pressure was 30.5 +/- 1.7 mm Hg. CONCLUSIONS: Unilateral double lobar lung transplantation was technically possible and associated with satisfactory early pulmonary function in a canine experimental model. PMID- 14762368 TI - Heart transplantation in patients with previous Fontan operations. AB - OBJECTIVE: The clinical features and outcomes of patients undergoing heart transplantation after a failed Fontan operation are still debated. The aim of this study was to retrospectively evaluate our experience in 14 patients undergoing heart transplantation after previous Fontan-type operations. METHODS: From 1990 to 2002, 14 patients underwent heart transplantation in our institution after a previous Fontan procedure. The mean age at the time of the Fontan operation and at transplantation was 7.3 +/- 2.8 and 17.2 +/- 6.3 years, respectively. The indication for transplantation was protein-losing enteropathy in 7 patients, arrhythmia with ventricular dysfunction in 5 patients, and heart failure in 2 patients. All patients received basic immunosuppressive therapy with cyclosporine (INN: ciclosporin) and azathioprine without induction therapy or maintenance steroids. RESULTS: Two hospital deaths occurred: one patient died on the fifth postoperative day of graft failure, and the second died on the 17th postoperative day after an acute neurologic event. Two patients died later, one 23 months after transplantation of acute rejection and the other after 90 months of chronic rejection and endocarditis. One patient underwent successful reintervention 2 years after heart transplantation for pulmonary vein obstruction. The 10 surviving patients are in New York Heart Association class I, with a mean follow-up of 64.5 +/- 42 months. One of them was delivered of a healthy baby 5 years after transplantation. Patients with protein-losing enteropathy reached a normal protein level within a mean of 10 months (range, 6 18 months) after transplantation. Four patients required a temporary administration (3-6 months) of oral steroid therapy for recurrent rejection episodes. Currently, 7 patients are taking cyclosporine, and 3 are taking cyclosporine and azathioprine. The actuarial survival at 1, 5, and 10 years was 86% +/- 9%, 77% +/- 12%, and 62% +/- 17%, respectively. CONCLUSION: Heart transplantation is a good option for patients with a failing Fontan operation. We documented the reversibility of protein-losing enteropathy in all patients. No mortality caused by surgical complications was observed. PMID- 14762370 TI - Clinical-pathologic conference in general thoracic surgery: Disseminated nocardiosis presenting as Pancoast syndrome. PMID- 14762371 TI - Partial atrioventricular canal defect with cor triatriatum sinister: report of three cases. PMID- 14762372 TI - Isolated pulmonary arteriovenous malformations requiring anatomic resection. PMID- 14762373 TI - Combined aortic arch repair and arterial switch without coronary reimplantation. PMID- 14762374 TI - Repair of aortic arch coarctation and innominate artery aneurysm with branched Dacron graft. PMID- 14762375 TI - Rescue of a case of ventricular septal defect with Eisenmenger syndrome using an extracorporeal membrane oxygenator. PMID- 14762376 TI - Thymic carcinoid combined with myasthenia gravis. PMID- 14762377 TI - Transposition of the lower pulmonary vein for further mobilization in carinal reconstruction after induction therapy for lung cancer. PMID- 14762378 TI - Pulmonary mucormycosis in a patient with chronic obstructive pulmonary disease: diagnosis by fine needle aspiration cytology. PMID- 14762379 TI - Pulmonary artery pseudoaneurysm in a patient with Behcet disease. PMID- 14762380 TI - Methylene blue for lithium-induced refractory hypotension in off-pump coronary artery bypass graft: report of two cases. PMID- 14762381 TI - Pathologic findings of aortic redissection after glue repair of proximal aorta. PMID- 14762382 TI - Left atrial dissection after mitral operation mimicking severe mitral regurgitation. PMID- 14762383 TI - Penetrating injury of the heart by a nail gun. PMID- 14762384 TI - An unusual complication after chordal sparing mitral valve replacement. PMID- 14762385 TI - Aortic dissection without Marfan's syndrome in ankylosing spondylitis. PMID- 14762386 TI - Total magnetic vascular coupling for arterial revascularization. PMID- 14762387 TI - Limitations on the role of vacuum-assisted closure in cardiac surgery. PMID- 14762389 TI - Heterotopic right heart transplantation. PMID- 14762388 TI - Reporting of clinical trials of analgesia. PMID- 14762390 TI - The radial artery: neither gold, nor silver, but bronze? PMID- 14762391 TI - Angiotensin-converting enzyme inhibitors predispose to hypotension refractory to norepinephrine but responsive to methylene blue. PMID- 14762392 TI - Modified Fontan procedure in adults. PMID- 14762393 TI - Mitral valve surgery with extensive calcification of the annulus. PMID- 14762395 TI - Orthopaedics: gene therapy's dark horse. PMID- 14762394 TI - Early failure of bovine jugular vein conduits. PMID- 14762396 TI - Gene therapy progress and prospects: cancer gene therapy using tumour suppressor genes. AB - Targeting tumour suppressor gene pathways is an attractive therapeutic strategy in cancer. Since the first clinical trial took place in 1996, at least 20 other trials have investigated the possibility of restoring p53 function, either alone or in combination with chemotherapy, but with limited success. Other recent clinical trials have sought to harness abnormalities in the p53 pathway to permit tumour-selective replication of adenoviral vectors such as dl1520 (Onyx-015). Other tumour suppressor genes, such as retinoblastoma (Rb) and PTEN (phosphatase, tensin homologue, deleted on chromosome 10), are the targets for imminent clinical trials, while microarray technologies are revealing multiple new genes that are potential targets for future gene therapy. PMID- 14762397 TI - Reciprocal regulation of the nuclear factor of activated T cells and HIV-1. AB - The human immunodeficiency virus type 1 (HIV-1) has evolved to coordinate its replication with the activation state of the host CD4T cell. To this end, it taps into major host cell signaling pathways and their associated transcription factors. Of these, T-cell activation and the transcription factor NF-kappaB, respectively, have become the best-studied examples. The past several years have revealed compelling evidence that another transcription factor family involved in T-cell activation, the nuclear factor of activated T cells (NFAT), plays an important role in the regulation of HIV-1. Major advances have been made in our understanding of the interaction of HIV-1 with this intriguing transcription factor. The duplicated NF-kappaB binding sites in the HIV-1 enhancer surprisingly also bind NFAT proteins and appear to be the most important targets for NFAT transactivation of the HIV-1 long terminal repeat. The crystal structure of NFAT1 bound to one of these duplicated sites was solved recently. Interestingly, it showed that NFAT1 binds to this site as a homodimer and occupies the core of the NF-kappaB site, suggesting mutually exclusive binding and alternate transactivation by these two factors. NFAT also regulates HIV-1 infection indirectly, as it can relieve a block to reverse transcription in quiescent T cells. In turn, HIV-1, and particularly its Tat and Nef gene products, can upregulate NFAT expression and activity. This reciprocal regulation between virus and transcription factor potentially creates a positive feedback loop, which may facilitate the establishment of early HIV-1 infection and, later, the transition from latent to productive infection. The immunosuppressive drug cyclosporin A (CsA) inhibits NFAT activity and thus represents a potential treatment for HIV-1 infection. Recent small-scale clinical trials have yielded optimistic results, suggesting roles for CsA after organ transplantation in HIV-1+ individuals and as adjunct treatment in stable early HIV-1 infection. PMID- 14762398 TI - Dissociation of disease susceptibility, inflammation and cytokine profile in lmr1/2 congenic mice infected with Leishmania major. AB - Severity of disease caused by Leishmania major depends on the genetics of the host. Early induction of T helper cell type 1 (Th1)-type responses in resistant C57BL/6 mice and T helper cell type 2 (Th2) in susceptible BALB/c mice is thought to determine cure or disease respectively. We have mapped three loci that confer susceptibility or resistance upon congenic mice on the C57BL/6 or BALB/c backgrounds. Here we examine the histopathology and production of interleukin 4 (IL-4) and interferon gamma (IFN-gamma) in the skin and draining lymph nodes in the congenic and parental mice. We show an evolving granuloma with a staged infiltration of inflammatory cells, but no difference between the groups. As an indication of an early-polarised Th1/Th2 response we measured IFN-gamma and IL-4 in the lymph nodes and found no difference between any of the mice during the first 48 h. During infection, the level of IL-4 correlated with the lesion size, indicating that IL-4 reflects the disease severity rather than controls it. Considering this effect, B6.C(lmr1,lmr2) mice had similar cytokine levels to the parental C57BL/6 mice despite increased susceptibility and C.B6(lmr1,lmr2) were similar to BALB/c despite increased resistance. We conclude that the lmr loci affect disease severity by a mechanism independent of conventional helper T-cell responses. PMID- 14762399 TI - A hand to hold: communication during cataract surgery. PMID- 14762400 TI - Growth factors and protein kinase C inhibitors as novel therapies for the medical management diabetic retinopathy. AB - Diabetic retinopathy is a leading cause of acquired visual loss. Current treatment modalities are not effective in all cases and may have side effects. Investigation of the biochemical basis of diabetic retinopathy suggests that future treatments may reverse or halt the progression of diabetic retinopathy, or actually prevent the development of diabetic retinopathy. Pharmacological manipulation of protein kinase C and various growth factors may form the basis of future treatments for diabetic retinopathy. PMID- 14762401 TI - Management of paediatric traumatic cataract by epilenticular intraocular lens implantation: long-term visual results and postoperative complications. AB - PURPOSE: This study was carried out to evaluate the postoperative visual results and complications following epilenticular intraocular lens implantation in paediatric traumatic cataracts. MATERIALS AND METHODS: A total of 30 patients undergoing epilenticular intraocular lens implantation for traumatic pediatric cataract were evaluated prospectively for visual results and complications. After cataract formation was the prime area of interest. Follow-up of patients ranged from 2 to 8 years (mean 5 years). In all, 90% of eyes had a visual acuity of 6/60 or worse at the time of presentation. RESULTS: In total, 80% of our patients achieved a visual acuity of 6/12 or better postoperatively after a mean follow-up of 5 years. None of our patients had after-cataract formation and the visual axis remained clear until the last follow-up in all the patients. CONCLUSION: Epilenticular intraocular lens implantation in paediatric traumatic cataracts is associated with favourable anatomic and visual results. PMID- 14762402 TI - Hepatitis C virus-associated keratitis. AB - AIM: To study the association between chronic hepatitis C virus (HCV) and Mooren's type keratitis. METHOD: A total of 50 patients with chronic HCV were screened for any evidence of corneal ulceration. Detailed ocular examination was conducted by slit-lamp biomicroscopy. Patients with history of trauma to the eye or previous herpetic keratitis were excluded from the study. RESULTS: There were 37 males and 13 females. The age of the patients ranged from 10 to 70 years. There was no evidence of Mooren's ulcer in any of our patients. CONCLUSION: No association between chronic HCV and Mooren's ulcer was found in our study. Screening therefore in such cases is not necessary. PMID- 14762403 TI - Accuracy of the clinical diagnosis of chalazion. AB - PURPOSE: A chalazion, localized lipogranulomatous inflammation of the eyelid, may simulate various eyelid lesions. This study was conducted to determine the accuracy of the clinical diagnosis of chalazion and demonstrate the importance of histopathological confirmation of the diagnosis. METHODS: Histopathological diagnoses of 1060 cases with the clinical diagnosis of chalazion, submitted to the Henry C Witelson Ophthalmic Pathology Laboratory and Registry between September 1993 and December 2001, were retrospectively evaluated. Discrepancies between clinical and histopathological diagnoses were classified. RESULTS: A total of 1033 (97.4%) of the 1060 cases were clinically diagnosed as primary and the remaining 27 (2.6%) as recurrent chalazions. Agreement was noted between clinical and histopathological diagnoses in 992 (93.6%) cases. Of the 68 (6.4%) clinically misdiagnosed cases, 15 (1.4%) were found to be malignant, two (0.2%) premalignant, and 51 (4.8%) benign conditions. Sebaceous cell carcinoma was the most commonly missed malignancy (12 cases, 1.1%) followed by basal cell carcinoma (three cases, 0.3%). Premalignant lesions, which masqueraded as chalazion, were chronic inflammation with cellular atypia and mitotic figures (two cases, 0.2%). Of these 17 cases with premalignant and malignant histopathologies, only six (35.3%) had a clinical diagnosis of recurrent chalazion, whereas the others (64.7%) were primary cases. Of the various benign conditions that were misdiagnosed as chalazion, different types of chronic inflammation (24 cases, 2.2%) were the most frequent. CONCLUSIONS: A number of different benign, premalignant, and malignant conditions may clinically masquerade as a chalazion. Delayed diagnosis and treatment of sebaceous cell carcinoma, which is the most frequently missed malignancy, may be life threatening for the patient. Therefore, all chalazion specimens, primary or recurrent, should be submitted for histopathological examination. PMID- 14762404 TI - Efficacy of photodynamic therapy in circumscribed choroidal haemangioma. AB - PURPOSE: To report efficacy of photodynamic therapy (PDT) in the treatment of three cases of juxtafoveal circumscribed choroidal haemangioma. METHODS: Data on three patients (two primary, and one failed TTT) treated with verteporfin, 6 mg/m2 given as i.v. infusion over 10 min. Diode laser (690 nm) with an intensity of 600 mW/cm2 for 83 s (50 mJ/cm2) was applied 5 min after completion of infusion. Overlapping multiple spots (2500 microm) were applied to cover the entire surface of the tumour. The mean pretreatment tumour size was 7 mm (base) x 2.2 (thickness) mm. Periodic follow-up with ophthalmoscopy, ultrasonography, and angiographic studies was performed. RESULTS: All three cases showed complete regression of the tumour with resolution of subretinal fluid, flattening of tumour, and absence of choroidal vasculature on ICG. The visual acuity either improved or remained stable in all three cases. The overlying retinal vessels remained unaffected. There were no ocular or systemic complications. CONCLUSIONS: PDT is an effective treatment for management of juxtafoveal circumscribed choroidal haemangioma. PMID- 14762405 TI - Heidelberg retina tomograph in ocular Behcet's disease. AB - PURPOSE: To compare the optic disc topography of patients with ocular Behcet's disease (BD) with age-matched controls, using Heidelberg retina tomograph (HRT). METHODS: This study included 32 patients (51 eyes) with ocular BD (active and/or inactive), 38 patients (74 eyes) with nonocular BD, and 62 normal subjects (62 eyes). All patients and control group underwent complete ophthalmologic evaluation. Intraocular pressure was less than 22 mmHg in patients and in the control group. The optic nerve heads of all subjects were imaged with the HRT (software 2.01a-M). The following stereometric parameters were calculated for each patient: disc area, cup area, cup/disc area ratio, rim area, height variation contour, cup volume, rim volume, mean cup depth, cup shape measure, mean RNFL thickness, and RNFL cross-sectional area. Differences among the groups were evaluated by Kruskal-Wallis variance analysis. When the Kruskal-Wallis test revealed a significant difference between groups, multiple comparison tests were used to find out which groups differed from which others. RESULTS: The mean disc area was significantly smaller (P<0.05) in patients with ocular and nonocular BD. The mean cup area, mean cup depth, and mean cup volume were significantly smaller (P<0.05) in patients with ocular BD. No significant differences were found between the groups in terms of the other stereometric parameters (P>0.05). CONCLUSION: A small disc and cup may be an additional risk factor for retinal vaso-occlusion in ocular Behcet's disease. PMID- 14762406 TI - Patient communication during cataract surgery. AB - PURPOSE: Most cataract surgery is now performed under local anaesthesia on a day case basis. As patients are fully conscious during the procedure, it is important that they remain still. There are a variety of reasons why patients may need to move, and it is important that the surgeon is made aware that this may happen. Some centres offer a nurse's hand as a means of perioperative patient communication. We sought to study the safety and efficacy of using an electronic patient-controlled alert device. METHOD: We compared hand-holding with the use of a patient alert device, and with both communication methods at the same time, on 150 subjects undergoing cataract surgery under local anaesthesia. Assessment of pre- and postoperative state anxiety was undertaken and patients' satisfaction with the communication strategies was assessed. RESULTS: There was a significant difference between pre- and postoperative state anxiety for each group (P<0.001) but no significant differences in pre-, peri-, or postoperative state anxiety between groups. There were no significant differences in confidence, pain, understanding, satisfaction, memory, and reassurance between the three groups. A total of 46% of all patients reported experiencing one or more of the potential problems enquired about, during the operation. Significant correlations were also identified between some of the psychological variables investigated. CONCLUSIONS: An electronic patient alert device is as effective a means of perioperative patient communication as holding a nurse's hand, during cataract surgery under local anaesthesia. It is safe, reassuring and it allows patients to communicate directly with the surgeon. PMID- 14762407 TI - Incidence of eye cancer in Taiwan: an 18-year review. AB - PURPOSE: To describe the incidence and histologic patterns of eye cancers in Chinese in Taiwan. METHODS: Beginning in 1979, cases of cancer in Taiwan were reported to the Taiwan National Cancer Registry. Information on all Chinese patients diagnosed with eye malignancies under the International Classification of Disease, Ninth Revision, site 190, was retrieved for analysis from the data bank of the Taiwan National Cancer Registry. RESULTS: In all, 733 cases with primary eye cancers were identified from 1979 to 1996, an 18-year period. The average annual age-standardized incidence of eye cancers was 2.46 per million population (2.57 for male and 2.33 for female). For cases less than 15 years of age, the most common eye malignancy was retinoblastoma (86.0%), followed by rhabdomyosarcoma (3.9%) and lymphoma (2.8%). For cases 15 years of age or older, the most common eye malignancy was melanoma (28.6%), followed by squamous cell sarcoma (21.0%) and lymphoma (20.8%). CONCLUSIONS: The time trends of the incidence of eye cancers were relatively stable over the 18-year period in Taiwan. Retinobalstoma, melanoma, and lymphoma were the three most common eye cancers in this Chinese population. PMID- 14762408 TI - Aberrant facial nerve regeneration (AFR): an under-recognized cause of ptosis. AB - INTRODUCTION: Aberrant facial nerve regeneration (AFR) following facial nerve palsy may give rise to ptosis because of increased orbicularis tone. We describe a series of patients presenting with ptosis where the underlying aetiology of AFR was often not recognized by the referring clinicians. METHODS: Retrospective case review. RESULTS: A total of 15 cases with ptosis, secondary to AFR, were seen at the Royal Adelaide Hospital Oculoplastic Clinic between 2000 and 2002. Of these, 10 (67%) were referred by general ophthalmologists. Ptosis was the only reason for referral in 11 patients (73%) and features of AFR or a past history of facial nerve palsy were not mentioned in seven referrals (overall 46%). All patients reported a previous facial palsy. The palpebral aperture was reduced on the affected side with reduction in both upper and lower margin reflex distance (MRD) by a mean of 1.5+/-0.7 mm (P<0.001) and 1.0+/-0.3 mm (P<0.001), respectively. The orbicularis tone was increased and strength reduced on the affected side in all patients. However, none had lagophthalmos. Signs of AFR were demonstrated in all patients with either an increase in ptosis or eyelid closure on the affected side during cheek puffing. CONCLUSION: Patients with AFR following facial nerve palsy may present with ptosis without recognition of the underlying aetiology. Signs of AFR ptosis include a decreased palpebral aperture with a reduced upper and lower MRD. The diagnosis can be established with demonstration of an increase in ptosis during cheek puffing. Recognition of AFR is important in these cases owing to the implications for management and post-operative patient satisfaction. PMID- 14762409 TI - Effect of cataract extraction and intraocular lens implantation on nerve fibre layer thickness measurements by scanning laser polarimeter (GDx) in glaucoma patients. AB - PURPOSE: To assess the influence of visually significant cataract on the measurement of nerve fibre layer thickness by scanning laser polarimetry (GDx) in glaucoma patients undergoing phacoemulsification cataract extraction. METHOD AND SUBJECTS: All subjects with primary glaucoma participating in a prospective trial of glaucoma surgery who subsequently underwent cataract extraction were eligible. A single trained observer using the GDx nerve fibre layer analyser (LDT) performed pre- and post-operative measurements of nerve fibre layer thickness (NFLT). NFLT parameters, best-corrected LogMAR visual acuity, and automated visual fields were assessed before and after phacoemulsification cataract extraction with implantation of an acrylic intraocular lens. RESULTS: A total of 49 subjects were assessed: 22 (45%) had POAG and 29 (55%) PACG; all were Asian (36 (73%) were Chinese), with mean age 67.1 (+/-7.6 SD) and mean 'LOCS III' lens opacity grading 11.4 (+/-3.1 SD). Visual acuity significantly improved (mean LogMAR 0.5 vs 0.15, P<0.0001). Corrected pattern standard deviation (6.1 vs 6.4, P=0.2) and mean deviation (-17.7 dB vs -17.0 P=0.91) were little changed after cataract removal. Pseudo-phakic measurements of NFLT were significantly different from pre-op values. Measures of absolute thickness (including the average thickness, ellipse, ellipse average, superior and inferior averages, superior integral) were significantly greater than preoperative values (all P<0.01), whereas ratios and measures of symmetry (symmetry, superior/nasal) were unchanged (all P>0.1) and 'the number' was smaller (P=0.04). Differences in measured NFLT were most strongly correlated with posterior subcapsular cataract (average thickness, P=0.01). CONCLUSIONS: Removal of cataract resulted in greater absolute measurements of NFLT but ratio values were unchanged. Scanning laser polarimetry measurements can change significantly after cataract extraction. New baseline measurements may be required. PMID- 14762410 TI - Effect of video display on the grading of diabetic retinopathy. AB - PURPOSE: Digital images are increasingly being used in ophthalmology. These may be viewed either on thin-film transistor (TFT) or on cathode-ray tube (CRT) displays. However, there is little data showing which is superior. In this study, we compared the performance of CRT and TFT displays for grading of both compressed and uncompressed images of diabetic retinopathy. METHODS: A total of 49 35 mm transparencies of diabetic retinopathy were scanned and compressed. The images comprised 17 with no retinopathy, eight with background, five with preproliferative, and 19 with proliferative retinopathy. Four levels of compression were used: 0, 70, 80, and 90%. A total of 196 randomised images were presented to two masked graders using both TFT and CRT displays under uniform lighting conditions, 2 months apart. The grade of retinopathy was assessed. Statistical analysis of grading accuracy was performed using receiver operator characteristic curves of sensitivity and specificity and the Stuart-Maxwell test for paired, nonparametric data. RESULTS: Both displays showed high sensitivity and specificity for the detection of any retinopathy. For the specific grade of retinopathy, the CRT performed slightly better with a sensitivity of 0.80 for uncompressed images, compared with 0.75 using the TFT. Compression reduced these sensitivities to 0.73 on the CRT and 0.63 on the TFT. Grading of uncompressed images magnified to four times their original size was more accurate on the TFT. CONCLUSIONS: Grading on both displays met sensitivity and specificity criteria proposed by Diabetes UK (formerly British Diabetic Association) for screening of diabetic retinopathy. The CRT generally performed slightly better than the TFT in relation to the detection of the specific grade of retinopathy. PMID- 14762411 TI - Comparison between standard Goldmann applanation prism and disposable applanation prism in tonometry. AB - BACKGROUND: Disposable devices are increasingly becoming the preferred choice where possible in contact medical equipment. AIM: To evaluate the accuracy of the disposable applanation tonometer head as a potential substitute to the standard Goldmann applanation head. METHODS: The study was prospective. The intraocular pressure recordings in 80 eyes of 42 patients were compared using the disposable and standard Goldmann applanator heads. The Bland and Altman method of assessing agreement between two methods of clinical measurement was used in the analysis. RESULTS: The difference in the readings between the two types of tonometer heads was highly variable (mean difference=0.78 mm Hg, range=-1 to 11 mm Hg). This was because of the distortions on the applanating surface of the disposable device. When the readings associated with the defective heads were excluded, very strong agreement was obtained (mean=0.07 mm Hg, range=-1 to 2 mm Hg). CONCLUSION: Good agreement with standard Goldmann applanation is achieved with the disposable heads except where surface distortions induce significant errors. Careful inspection to ensure well-structured disposable units is imperative in disposable applanation tonometry. PMID- 14762412 TI - Management of endophthalmitis with retained intraocular foreign body. AB - AIMS: To assess the impact of primary injection of intravitreal antibiotics and delayed pars plana vitrectomy with removal of intraocular foreign body (IOFB) in patients with clinical features of bacterial endophthalmitis and retained IOFB. METHODS: Retrospective review of all patients with clinical features of infective endophthalmitis and a retained IOFB who had immediate injection of intravitreal antibiotics and delayed pars plana vitrectomy with removal of IOFB in two vitreo retinal centres during 1995-2001. Nine patients were identified and minimum follow-up was 3 months. RESULTS: Four of the nine patients had a final visual outcome of 6/18 or better. One patient developed total retinal detachment. CONCLUSIONS: The current series suggests that immediate injection of intravitreal antibiotics with delayed removal of IOFB in eyes with clinical features of infective endophthalmitis and a retained IOFB is a possible alternative to immediate removal of IOFB. This management may be associated with preservation of the eye and restoration of useful visual acuity. PMID- 14762413 TI - Illuminated artificial orbit for the training of vitreoretinal surgery in vitro. AB - PURPOSE: Demonstrations and practical experiments are important constituents of the teaching of surgical principles and techniques to residents in ophthalmology. The objective was to develop an eye-support device for experimental and didactic use during vitreoretinal microsurgery in vitro. METHODS: Various eye-support devices with different light sources and illumination intensities were designed and tested. The main components of the final prototype are a high-intensity halogen lamp, a flexible light guide, a cylinder with a mirror inside, and a replaceable, transparent eye support with a ring-adapter. Light from the point source is reflected via the light guide into the cylinder and through the eye support transsclerally into the eye. RESULTS: The device illuminates the retina and the vitreous body sufficiently. It accentuates the transparent appearance of the retina and makes the three-dimensional structure more visible, for example, in case of retinal detachment. Subretinal tissue can be visualized and differentiated more precisely. CONCLUSIONS: The device is useful, in the teaching of residents and in experimental vitreoretinal surgery. PMID- 14762414 TI - Endoscopic visualisation to aid deep anterior lamellar keratoplasty. AB - AIMS: The aim of this study was to assess the value of endoscopy during deep anterior lamellar keratoplasty (DALK) by visualising the posterior cornea. This allows the surgeon to determine whether air injection had succeeded in stripping Descemet's membrane and endothelium from the posterior corneal stroma. METHODS: Four whole globes for research were obtained from the Florida eye bank with consent. A 2 mm incision was placed at the limbus and the endoscope was introduced through this into the anterior chamber. A 26-gauge needle was introduced into the cornea with the bevel positioned as deep as possible and air injected into the corneal stroma. Air was injected until the whole cornea became opaque and repeated air injections were made even after an opaque cornea was noted. The endoscopic camera was used to visualise the posterior corneal surface during this procedure. RESULTS: The view of the posterior corneal surface was clear and introduction of the probe did not interfere with the air dissection. In all four eyes, despite ease of air injection and diffuse corneal air infiltration, no large air bubble dissection of Descemet's membrane from adjacent stroma occurred. Instead multiple blistering of the posterior corneal surface could be seen. CONCLUSIONS: Endoscopy provides an effective tool to visualise the posterior corneal surface during DALK, using air dissection. This technique may become a standard adjunctive procedure during DALK. PMID- 14762415 TI - A lash foreign body complicating subtenon's anaesthesia. PMID- 14762416 TI - YAG laser capsulotomy, an unusual complication. PMID- 14762417 TI - Refractive error following cataract extraction with the implantation of a standard power intraocular lens in a rural African blindness prevention programme. PMID- 14762418 TI - Subretinal Pseudomonas abscess in a patient with bronchiectasis. PMID- 14762419 TI - Regression of aneurysmal dilatations in a case of idiopathic retinal vasculitis, aneurysms and neuroretinitis (IRVAN) associated with allergic fungal sinusitis. PMID- 14762420 TI - Pseudoxanthoma elasticum and nonarteritic anterior ischaemic optic neuropathy. PMID- 14762421 TI - Atypical acute retinal necrosis (ARN) responding to late treatment. PMID- 14762422 TI - Bilateral rhegmatogenous retinal detachment secondary to retinal dialyses associated with multiple retinal breaks. PMID- 14762423 TI - Acute bilateral blindness in meningeal carcinomatosis. PMID- 14762424 TI - Use of pilocarpine following hyphaema-related ocular hypertension. PMID- 14762425 TI - Abnormal lens shape on CT in a patient with Aniridia. PMID- 14762426 TI - Drug-induced leukopenia in a diabetic patient with larvate endophthalmitis. PMID- 14762427 TI - Traumatic and postoperative hyphaema in a patient with sickle cell trait. PMID- 14762428 TI - Bilateral periocular swelling in Sweet's syndrome. PMID- 14762429 TI - Optic disc haemorrhage following frontal head trauma. PMID- 14762430 TI - Calcification in Schwannoma of the lacrimal gland region. PMID- 14762431 TI - Ocular tuberculosis with angle granuloma. PMID- 14762432 TI - Traumatic rupture of the lateral rectus. PMID- 14762433 TI - Bilateral endophthalmitis due to self-mutilation. PMID- 14762434 TI - Exudative retinal detachment following central retinal vein occlusion. PMID- 14762435 TI - One-stop cataract clinics: feasible but flawed? PMID- 14762436 TI - Retinal tear following closed trauma with a toy balloon. PMID- 14762437 TI - Postoperative infection and subtenon anaesthesia. PMID- 14762438 TI - Progressive glaucoma in patients with Alzheimer's disease. PMID- 14762439 TI - Smad4-independent regulation of p21/WAF1 by transforming growth factor-beta. AB - The transforming growth factor-beta (TGF-beta)-Smad signaling pathway inhibits the growth of human epithelial cells and plays a role in tumor suppression. The Smad4 gene is mutated or deleted in 50% of pancreatic cancers. In this study, the Smad4-null pancreatic cancer cell line BxPC-3 was transfected with either the Smad4 expression vector or the empty vector and incubated in the presence or absence of TGF-beta. The cells were analysed using a cDNA microarray, which included 2280 named genes to screen for target genes regulated by TGF-beta in either a Smad4-dependent or -independent manner. The microarray and subsequent quantitative RT-PCR analysis demonstrated that the Smad4-independent and dependent signaling pathways driven by TGF-beta upregulated only one of the 2280 genes, respectively, suggesting that Smad4-independent signaling downstream of TGF-beta might be as widespread as Smad4-dependent signaling. In this study, we demonstrated that the cyclin-dependent kinase inhibitor p21/WAF1, which has been considered the major effector of the Smad-dependent growth inhibitory signal of TGF-beta, is upregulated in a Smad4-independent manner. The upregulation occurs through Smad2/3-dependent transcriptional activation of the p21/WAF1 promoter region. These results suggest a novel mechanism of gene regulation, that is, a novel signal mediator other than Smad4. PMID- 14762440 TI - Overexpression of estrogen receptor-alpha gene suppresses gap junctional intercellular communication in endometrial carcinoma cells. AB - Stimulation of the endometrium by estrogens without the differentiating effect of progestins is the primary etiological factor associated with the development of endometrial hyperplasia and adenocarcinoma. However, the correlation between sex steroids and gap junctional intercellular communication (GJIC), which is considered to play an important role in the control of cell growth and differentiation, is not well known in endometrial carcinoma. In this study, we focused on the influence of estrogen and its receptor in connexin (Cx) expression and GJIC in endometrial carcinoma cells, established stable clone IK-ER1 overexpressing ER-alpha to transfect the expression vector and analysed them in various hormonal conditions. The growth of IK-ER1 was accelerated by 17beta estradiol and the acceleration of the 5-bromo-25-deoxyuridine labeling index was observed. GJIC was assayed by scoring the number of dye-coupled cells after microinjection of single cells with Lucifer-Yellow, and subcellular localization of Cx26 and Cx32 was analysed by immunocytochemistry. In the presence of estradiol, dye-coupled cells of IK-ER1 were significantly reduced compared to those without estradiol and the reduction was completely inhibited by adding ICI182.780, a pure antiestrogen substrate. Cxs were detected as only small spots by immunocytochemistry, and Western blotting showed that the expression was decreased. These results suggest that activation of ER-alpha by estrogen results in tumor progression by stimulating cell growth and suppressing GJIC via suppression of the expression of Cxs in endometrial carcinogenesis. PMID- 14762441 TI - IRF-1 expression induces apoptosis and inhibits tumor growth in mouse mammary cancer cells in vitro and in vivo. AB - Interferon regulatory factor-1 (IRF-1) is a nuclear transcription factor that mediates interferon and other cytokine effects and appears to have antitumor activity in vitro and in vivo in cancer cells. We have constructed a recombinant adenoviral vector (Ad-IRF-1) that infects mammary cells with high efficiency and results in high levels of functional IRF-1 protein in transfected cells. Overexpression of IRF-1 in two mouse breast cancer cell lines, C3-L5 and TS/A, resulted in apoptosis in these cell lines as assessed by Annexin V staining. The involvement of caspases was confirmed by significant inhibition of apoptosis by a caspase inhibitor, and by demonstration of caspase-3 activity, cleavage of caspase-3, and PARP cleavage. Interestingly, the growth of nonmalignant breast cell lines C127I and NMuMG did not appear to be inhibited by IRF-1 overexpression. Suppression of growth for breast cancer cell lines in vivo was demonstrated by both preinfection of breast cancer cells ex vivo and by intratumoral injection of Ad-IRF-1 into established tumors in their natural hosts. The mechanism of apoptosis may involve the transcriptional upregulation of bak, caspase-8, and caspase-7 expression. These data support the antitumor potential of IRF-1 and the use of agents that increase IRF-1 in breast cancer. PMID- 14762443 TI - Incidence and prognostic value of respiratory events in acute leukemia. AB - Acute respiratory failure and infectious pneumonia are the major causes of death during induction chemotherapy of acute leukemia. However, the causes, incidence and prognostic value of all respiratory events (REs) occurring in this context have never been assessed prospectively. We recruited 65 consecutive patients with newly diagnosed acute leukemia into a 1-year prospective study (December 2000 November 2001) to evaluate the incidence and prognostic value of these events. REs were frequent: 38 were recorded in 30 patients. There was a significant relationship between REs and pre-existing respiratory disease and/or smoking. REs were caused by infection in 34% of cases, by an established cause other than infection in 42% and had an undetermined cause in 24%. Poor early outcome (death within 45 days of starting induction chemotherapy) in patients experiencing an RE was independently associated with a >25/min respiratory rate (P=0.003) and the nonachievement of complete remission (CR) (P<0.0001). Predictors of overall survival in the entire patient population were the absence of CR (P<0.0001), REs (P=0.02) and a > or =2 performance status (P=0.03). In conclusion, REs are frequent during induction chemotherapy of acute leukemia and represent an independent prognostic factor of poor outcome, regardless of their cause. PMID- 14762442 TI - Notch and Schwann cell transformation. AB - Benign plexiform neurofibromas in NF1 patients can transform spontaneously into malignant peripheral nerve sheath tumors (MPNSTs). Although mutations in the p53 gene have been found in a subset of MPNSTs and mouse models support a role for p53 mutations in malignant conversion, we found that each of three Schwann cell lines derived from human MPNSTs possessed active p53. One of the lines expressed the Notch intracellular domain (NICD), indicative of ongoing Notch signaling. Consistent with a role in malignancy, NICD was able to transform primary rat Schwann cells. Transformation was robust--NICD-transduced cells generated tumors in nude rats--and was associated with the loss of markers associated with Schwann cell differentiation. These data suggest that aberrant Notch signaling may contribute to the conversion of benign neurofibromas to MPNSTs. PMID- 14762444 TI - Decision-making and quality of life in older adults with acute myeloid leukemia or advanced myelodysplastic syndrome. AB - Older patients with acute myeloid leukemia (AML) and advanced myelodysplastic syndrome (aMDS) must decide between receiving intensive induction chemotherapy (IC) or nonintensive chemotherapy/best supportive care (NIC). Little information exists about what factors influence treatment decisions and what quality of life (QOL) is associated with treatment choices. We prospectively examined 43 patients 60 years or older who were interviewed at diagnosis and periodically over 1 year. IC choice was associated with younger age (66 vs 76 years, P=0.01) and AML diagnosis, but not with performance status, comorbidities, or QOL. In total, 63% of all patients reported not being offered other treatment options despite physician documentation of alternatives. Patient and physician estimates of cure differed significantly: 74% of patients estimated their chance of cure to be 50% or greater, yet for 89% of patients physician estimates of cure were 10% or less. IC patients experienced decreased QOL at 2 weeks, but rebounded to baseline and to NIC levels by 6 weeks. Initial QOL is not associated with treatment choice in older AML and aMDS patients. Regardless of treatment choice, patients report not being offered treatment options and overestimate their chances of cure. In IC patients, QOL decreases during hospitalization but rebounds after discharge. PMID- 14762445 TI - Minimal residual disease cells in AML patients have an apoptosis-sensitive protein profile. PMID- 14762446 TI - CD34+ progenitors from MDS patients are unresponsive to SDF-1, despite high levels of SDF-1 in bone marrow plasma. PMID- 14762447 TI - Does umbilical cord care in preterm infants influence cord bacterial colonization or detachment? AB - OBJECTIVE: The purpose of the present study was to compare alcohol versus natural drying for umbilical cord care in preterm infants and to examine its effects on bacterial colonization and cord detachment. STUDY DESIGN AND METHODS: Eligible infants <34 weeks gestation admitted to the NICU were randomized to receive either umbilical cleansing with 70% isopropyl alcohol at each diaper change or natural drying. Umbilical stump cultures were performed at 12 to 24 hours, 72 hours, 7 days, and 14 days of age. RESULTS: A total of 109 infants were enrolled; 102 completed the study. Analyses revealed that the median cord detachment time was significantly shorter in the natural drying group compared to the alcohol group (13.0 versus 16.0 days; p=0.003). There were no cases of local umbilical infection in either group. CONCLUSION: Based on the present study, it appears that natural drying is a safe and effective means of umbilical cord care in preterm infants. PMID- 14762448 TI - Prenatal care in the Paso del Norte border region. AB - OBJECTIVES: The impact of a Medicaid-managed care system on access to prenatal care was investigated. STUDY DESIGN: Postpartum interviews and medical chart abstractions were conducted among 493 Hispanic women who reside on the El Paso Texas/Juarez Mexico border (the Paso del Norte region). Descriptive analysis identified barriers and facilitators to prenatal care. Logistic regression identified the impact of social and demographic characteristics on selected maternal and infant outcomes. RESULTS: The factors reported by these women as barriers to timely entry and sustaining enrollment in prenatal care were related to the availability of social support networks and affiliation with the Mexican/Hispanic culture (acculturation). Having Medicaid-managed care or other insurance was associated with receiving more adequate levels of prenatal care. Women who crossed the border to seek perinatal services were more likely to have infants who received higher levels of neonatal care (odds ratio 0.500; 95% CI [0.264, 0.946]). CONCLUSIONS: The promotion of preconceptional, prenatal, and family planning services is strongly recommended as a strategic, regional, public health intervention. PMID- 14762449 TI - An amniotic fluid index < or =5 cm within 7 days of delivery in the third trimester is not associated with decreasing umbilical arterial pH and base excess. AB - OBJECTIVE: To determine if an amniotic fluid index (AFI) < or =5.0 cm within 7 days of delivery in the third trimester is associated with decreasing umbilical arterial pH and base excess. STUDY DESIGN: Cases for this retrospective cohort study were all pregnancies > or =26 weeks with intact membranes and an AFI < or =5.0 cm within 7 days of delivery between 11/99 and 7/02. Multiple gestations, aneuploid, and anomalous fetuses were excluded. Controls with an AFI >5.0 cm within 7 days of delivery were matched to each case within 1 week by gestational age. For a control group with a mean+/-SD umbilical arterial pH of 7.26+/-0.07 and alpha=0.05, a sample size of 100 would have a power of 99% to detect a difference with a study group whose mean was 7.20. Data were compared using paired Student's t-test, Mann-Whitney, Fisher's exact, chi(2) and risk ratios with 95% confidence intervals. RESULTS: In all, 131 neonates with an AFI < or =5.0 cm were matched to 131 controls with an AFI >5 cm. There was no difference in gestational age (37.6+/-3.0, 37.7+/-3.0 weeks) or birth weight (2897+/-810, 2762+/-788 g). There was no difference in umbilical artery pH (7.25+/-0.07, 7.26+/-0.07) or base excess (-3.32+/-2.59, -2.83+/-2.45 mmol/l), even in small for gestational age (SGA) infants in both groups. There was no difference in the number of SGA neonates, 5-minute Apgar <7, respiratory distress syndrome, necrotizing enterocolitis, or neurologic morbidity. Linear regression showed no correlation between AFI and either umbilical arterial pH (r=-0.00047, SE=0.001, p=0.63) or base excess (r=-0.029, SE=0.037, p=0.428). CONCLUSION: An AFI < or =5.0 cm measured within 7 days of delivery in the third trimester is not associated with decreasing umbilical arterial pH and base excess. PMID- 14762450 TI - Determinants of intrapartum fetal death in a remote and indigent population in China. AB - OBJECTIVE: To explore the reasons for the high rate of intrapartum fetal death observed in a remote and indigent population in China. STUDY DESIGN: We conducted an epidemiologic analysis of determinants of intrapartum fetal death in a sample of 20,891 births in 18 hospitals participating in the Qingyuan Perinatal Surveillance System from January 1, 1997 to June 30, 1998. The main determinant examined was cesarean delivery; other determinants included mother's insurance status, residence, maternal age, infant's gender, parity, gestational age, birth weight, and obstetric complications. Rates of intrapartum fetal death within categories of various maternal and infant factors were first calculated and compared; adjusted odds ratios for intrapartum fetal death were then estimated by multiple logistic regression analysis. RESULTS: The intrapartum fetal death rate in this population was 5 per 1000 total births, which accounted for about one third of all fetal deaths. Compared with vaginal delivery, elective cesarean delivery was associated with a 100% (i.e., no intrapartum fetal death among 1572 elective cesarean deliveries) and emergency cesarean delivery with a 88% reduction, in intrapartum fetal death. Other significant determinants were related to access to obstetric care (i.e., insurance status and residence). CONCLUSION: Lack of access to quality obstetric care is the major determinant of intrapartum fetal death in this population. PMID- 14762451 TI - Congenital complete heart block: fetal management protocol, review of the literature, and report of the smallest successful pacemaker implantation. AB - Fetuses with complete heart block have an increased mortality with most deaths occurring in utero or during infancy. The cardiac evaluation of these fetuses is difficult since the ventricular rate is low and the heart is dilated. We have implemented a strategy that includes the biophysical profile, which assesses fetal well-being, in combination with the cardiovascular profile that assesses cardiac function and the circulation. We present two cases of fetal complete heart block in which early delivery was recommended due to worsening cardiovascular profile scores. Biophysical profile scores were normal. Both babies were successfully treated, despite having risk factors that predicted poor outcomes. We hypothesize that our management protocol initiated intervention before fetal compromise, hydrops, and myocardial damage occurred. We recommend an evaluation of heart function in addition to an assessment of fetal well-being in fetuses with complete heart block. Early delivery should be considered if there is evidence of distress and/or deteriorating cardiac function. PMID- 14762452 TI - Arterial oxygen fluctuation and retinopathy of prematurity in very-low-birth weight infants. AB - OBJECTIVE: To determine the influence of arterial oxygen fluctuation on development of threshold ROP. STUDY DESIGN: Retrospective study of 231 infants, < or =1500 g birth weight, who were admitted to Arkansas Children's Hospital NICU from January 1993 to June 1995. Fluctuation in partial pressure of dissolved arterial oxygen (PaO(2)) was expressed as coefficient of variation (CoV) for each infant. We investigated the relationship between CoV at three intervals and the risk of developing threshold ROP. RESULTS: The odds ratio (OR) of developing threshold ROP versus prethreshold ROP or less associated with a 10% increase in the CoV during the first 5 days of oxygen therapy was 1.44, and during the first 10 days was 1.51. When analysis was restricted to infants receiving 30 days of therapy, the OR during the first 5 days of therapy was 1.67, during the first 10 days was 1.82, and during days 11-30 was 1.68. CONCLUSIONS: Very-low-birth-weight infants experiencing fluctuating PaO(2) are at higher risk of threshold ROP. PMID- 14762453 TI - Cytomegalovirus-associated necrotizing enterocolitis in a preterm twin after breastfeeding. AB - Cytomegalovirus (CMV) in breast milk is transmitted to infants and may be associated with disease especially in preterm infants. We present a preterm twin with postnatally acquired CMV infection and evidence of CMV-associated necrotizing enterocolitis. PMID- 14762454 TI - High-frequency partial liquid ventilation in two infants. AB - Two infants on high-frequency oscillatory ventilation for chronic lung disease and severe respiratory failure, received a bolus of warmed and oxygenated perfluorodecalin up to residual functional capacity, followed by a continuous infusion of 6 ml/kg/hour. Our aim was to improve gas exchange without increasing ventilatory-induced lung injury. Heart rate, oxygen saturation, blood pressure, and TcPO(2)/TcPCO(2) were continuously monitored during treatment. Arterial blood gas was evaluated every 3 hours. Both patients showed improvement of gas exchange with a 13.6 and 12.5% reduction of oxygenation index, respectively. High frequency partial liquid ventilation is an experimental ventilation technique that could be considered as rescue treatment, to improve oxygenation in subjects with critical respiratory failure. This method could probably produce less damage, than other ventilation modes, to severely injured lungs. PMID- 14762455 TI - Iopanoic acid in the management of neonatal Graves' disease. AB - OBJECTIVE: Traditionally, neonatal thyrotoxicosis has been managed with antithyroid drugs and/or iodine as well as sedatives, propranol and digitalis when necessary. The purpose of this study was to evaluate the management of neonatal thyrotoxicosis using the radio-contrast agent iopanoic acid. METHODS: We managed five cases of neonatal thyrotoxicosis. All infants were treated initially with propranolol (1.7 mg/kg/day) and iopanoic acid 250 to 500 mg every third or fourth day. RESULTS: In all cases, clinical signs improved and T(3) and T(4) levels decreased dramatically within 24 to 72 hours. No toxic side effects were noted. CONCLUSION: Neonatal thyrotoxicosis can be managed successfully using iopanoic acid. Iopanoic acid is essentially free of side effects and need only be administered every 3 to 4 days. When administered until (transplacental) maternal TSI has been metabolized by the neonate, iopanoic acid maintains euthyroid status with no risk of hypothyroidism. With conventional therapy, propylthiouracil (PTU) must be administered three times a day. PTU also carries a significant risk of toxic side effects and a week or more of therapy is required to correct the hyperthyroid state and may induce hypothyroidism. PMID- 14762456 TI - Trisomy 14 mosaicism: a case report and review of the literature. AB - Trisomy 14 mosaicism is a rare chromosomal abnormality with distinct and recognizable clinical features. We describe two previously unreported abnormalities in this condition and delineate physical and psychomotor features and concerns for medical management. Trisomy 14 mosaicism should be suspected in individuals who have the features described herein, thus prompting cytogenetic evaluation of blood, and possibly other tissues for diagnosis. PMID- 14762457 TI - Embracing open access. PMID- 14762458 TI - Death in the family: the Medical Library Center of New York, 1960-2003. PMID- 14762459 TI - Being there: the library as place. AB - The value of the library as place is examined in this Janet Doe Lecture. The lecture, which is intended to focus on the history or philosophy of health sciences librarianship, presents an overview of the library as a place in society from ancient times to the present. The impact of information technology and changes in the methods of scholarly publication from print to digital are addressed as well as the role of the library as the repository of the written historical record of cultures. Functions and services of libraries are discussed in light of the physical library facility of the future. Finally, librarians are asked to remember the enduring values of librarianship in planning libraries of the future. PMID- 14762460 TI - Evaluating the effectiveness of clinical medical librarian programs: a systematic review of the literature. AB - OBJECTIVE: This study was undertaken to determine if a systematic review of the evidence from thirty years of literature evaluating clinical medical librarian (CML) programs could help clarify the effectiveness of this outreach service model. METHODS: A descriptive review of the CML literature describes the general characteristics of these services as they have been implemented, primarily in teaching-hospital settings. Comprehensive searches for CML studies using quantitative or qualitative evaluation methods were conducted in the medical, allied health, librarianship, and social sciences literature. FINDINGS: Thirty five studies published between 1974 and 2001 met the review criteria. Most (30) evaluated single, active programs and used descriptive research methods (e.g., use statistics or surveys/questionnaires). A weighted average of 89% of users in twelve studies found CML services useful and of high quality, and 65% of users in another overlapping, but not identical, twelve studies said these services contributed to improved patient care. CONCLUSIONS: The total amount of research evidence for CML program effectiveness is not great and most of it is descriptive rather than comparative or analytically qualitative. Standards are needed to consistently evaluate CML or informationist programs in the future. A carefully structured multiprogram study including three to five of the best current programs is needed to define the true value of these services. PMID- 14762461 TI - Research on the value of medical library services: does it make an impact in the health care literature? AB - OBJECTIVE: To evaluate the impact in the health care literature of research articles that provided evidence of the value of library services (including MEDLINE) as an element of quality health care. DATA SOURCES/SELECTION: Four research articles on the relationship between use of library services and quality health care were selected as "primary articles" from a MEDLINE search using appropriate Medical Subject Heading. Primary articles met the following criteria: written in English, reported research, related to clinical care, and published before 1995. DATA EXTRACTION: The technique of citation analysis was used to measure the impact of the primary articles on the subsequent literature. The number, authorship, type, and publication venue of articles citing the primary articles were determined using ISI Web of Science, MEDLINE, other electronic resources, and the citing articles themselves. For the 146 English-language citing articles, the article type (i.e., advocacy, instructional, research) was noted; and, for those that reported research, the use to which the author put the cited material was determined. RESULTS: The primary articles were cited more often than the average articles published that year in the same journals. At the time of the study each article had been cited almost every year since publication. Of the 146 citing articles written in English, 43% were written by librarians, 38% by physicians, 12% by librarians with physicians. The majority were published in medical journals, followed in order of decreasing frequency by the Bulletin of the Medical Library Association, information science journals, and health administration journals. CONCLUSIONS: The results of this study demonstrate that published research on the value of medical library services has an impact on the literature. These articles are read and cited and continue to be of value. PMID- 14762462 TI - Identifying and communicating the contributions of library and information services in hospitals and academic health sciences centers. AB - OBJECTIVE: This article introduces a systematic approach to identifying and communicating the value of library and information services (LIS) from the perspective of their contributions to achieving organizational goals. METHODS: The contributions of library and information services (CLIS) approach for identifying and communicating the value of LIS draws on findings from a multimethod study of hospitals and academic health sciences centers. RESULTS: The CLIS approach is based on the concept that an individual unit's value to an organization can be demonstrated by identifying and measuring its contributions to organizational goals. The CLIS approach involves seven steps: (1) selecting appropriate organizational goals that are meaningful in a specific setting; (2) linking LIS contributions to organizational goals; (3) obtaining data from users on the correspondence between LIS contributions and LIS services; (4) selecting measures for LIS services; (5) collecting and analyzing data for the selected measures; (6) planning and sustaining communication with administrators about LIS contributions; and (7) evaluating findings and revising selected goals, contributions, and services as necessary. CONCLUSIONS: The taxonomy of LIS contributions and the CLIS approach emerged from research conducted in hospitals and academic health sciences centers and reflect the mission and goals common in these organizations. However, both the taxonomy and the CLIS approach may be adapted for communicating the value of LIS in other settings. PMID- 14762463 TI - Information needs of gay, lesbian, bisexual, and transgendered health care professionals: results of an Internet survey. AB - OBJECTIVES: To obtain basic facts and considered opinions from health care professionals and students (nonlibrarian and librarian) about the information needs of gay, lesbian, bisexual, and transgendered (GLBT) health care professionals and their interactions with medical librarians. METHODS: The survey instrument was a Web-based questionnaire. A nonrandom sample of health care professionals and students (librarian and nonlibrarian) was obtained by posting messages to several large Internet electronic discussion groups (GLBT and general) and to randomly selected members of the Gay and Lesbian Medical Association. A total of 152 forms were analyzed with about 50% of the participants being GLBT persons. RESULTS: GLBT people have specific health information needs and concerns. More than 75% of medical librarians and students believed that GLBT persons have special information needs, with similar response rates by nonlibrarian health professionals and students. The delivery of services needs to be done with privacy and respect for the feelings of the patron. Major areas of need include the topics of health care proxy, cancer, adolescent depression and suicide, adoption, sexual health and practices, HIV infection, surrogate parenting, mental health issues, transgender health issues, intimate partner violence, and intimate partner loss. CONCLUSIONS: Most GLBT health care professionals desire GLBT-friendly health information services. Making GLBT oriented health information resources available on a library Web page and making an effort to show acceptance of cultural diversity through signs or displays would be helpful. Education directed toward instilling an awareness of GLBT persons may also be advisable. Most survey participants make some use of medical reference services and many find medical librarians to be very helpful and resourceful. PMID- 14762464 TI - Evaluation of five full-text drug databases by pharmacy students, faculty, and librarians: do the groups agree? AB - OBJECTIVES: The purpose of this study is to assess the usefulness of five full text drug databases as evaluated by medical librarians, pharmacy faculty, and pharmacy students at an academic health center. Study findings and recommendations are offered as guidance to librarians responsible for purchasing decisions. METHODS: Four pharmacy students, four pharmacy faculty members, and four medical librarians answered ten drug information questions using the databases AHFS Drug Information (STAT!Ref); DRUGDEX (Micromedex); eFacts (Drug Facts and Comparisons); Lexi-Drugs Online (Lexi-Comp); and the PDR Electronic Library (Micromedex). Participants noted whether each database contained answers to the questions and evaluated each database on ease of navigation, screen readability, overall satisfaction, and product recommendation. RESULTS: While each study group found that DRUGDEX provided the most direct answers to the ten questions, faculty members gave Lexi-Drugs the highest overall rating. Students favored eFacts. The faculty and students found the PDR least useful. Librarians ranked DRUGDEX the highest and AHFS the lowest. The comments of pharmacy faculty and students show that these groups preferred concise, easy-to-use sources; librarians focused on the comprehensiveness, layout, and supporting references of the databases. CONCLUSION: This study demonstrates the importance of consulting with primary clientele before purchasing databases. Although there are many online drug databases to consider, present findings offer strong support for eFacts, Lexi-Drugs, and DRUGDEX. PMID- 14762465 TI - From both sides now: librarians' experiences at the Rocky Mountain Evidence-Based Health Care Workshop. AB - The Colorado Health Outcomes (COHO) Department of the School of Medicine at the University of Colorado Health Sciences Center (UCHSC) coordinates the Rocky Mountain Evidence-Based Health Care (EBHC) Workshop, which has been held annually since 1999. The goals of the workshop include helping participants-physicians, pharmacists, health care policy makers, journalists and librarians-learn and apply skills for critically appraising medical research literature and for effective use of evidence-based information resources. Participants are encouraged to share ideas and to plan local services and instruction for those working in clinical settings. Each year, librarians from UCHSC Denison Memorial Library participate as faculty by teaching searching skills (PubMed, Cochrane Library, ACP Journal Club, etc.), providing support to small groups, and staffing two computer labs. In 2002, Denison Library received a National Network of Libraries of Medicine (NN/LM) MidContinental Region Impact Award to fund the attendance of three health sciences librarians from the MidContinental Region, an academic education librarian, a clinical medical librarian, and a department librarian. In this paper, the participating librarians share the lessons they learned about how health care practitioners approach evidence-based practice. The participating librarians also share how they incorporated these lessons into their support of evidence-based practice related to teaching about evidence-based resources, assisting health care practitioners with developing answerable questions, enhancing the clinician-librarian partnership, and assisting practitioners in selecting evidence-based resources for quick answers to clinical questions. PMID- 14762467 TI - Inventory of research methods for librarianship and informatics. AB - This article defines and describes the rich variety of research designs found in librarianship and informatics practice. Familiarity with the range of methods and the ability to make distinctions between those specific methods can enable authors to label their research reports correctly. The author has compiled an inventory of methods from a variety of disciplines, but with attention to the relevant applications of a methodology to the field of librarianship. Each entry in the inventory includes a definition and description for the particular research method. Some entries include references to resource material and examples. PMID- 14762466 TI - Information needs at the end of life: a content analysis of one person's story. AB - During the last phase of life, a person may need a variety of information to help her or him cope with dying and death. This article describes the nature of information needed during this stage. A content analysis was done of a book of conversations between a husband who was dying and his wife who is a grief counselor to determine his information needs. Four categories of needs were proposed, including physical, emotional, spiritual, and financial. Information needs germane to each category were identified. More research needs to be done by library and information science professionals to determine the information needs of people who are dying, as well as those of their families and the health professionals who care for them. PMID- 14762468 TI - Journal-citation-seeking behavior at two health sciences libraries. PMID- 14762469 TI - In a new element: medical librarians making patient education rounds. PMID- 14762470 TI - An examination of PubMed's ability to disambiguate subject queries and journal title queries. PMID- 14762471 TI - Proceedings, 103rd Annual Meeting Medical Library Association, Inc. San Diego, California, May 2-7, 2003. PMID- 14762473 TI - [The pharyngoesophageal transition and its mistery]. PMID- 14762474 TI - [Laryngopharyngeal structural analysis and its morphofunctional correlation with cricopharyngeal myotomy, botulinum toxin injection and balloon dilation]. AB - BACKGROUND: The cricopharyngeal muscle is of the skeletal type and, in this way, unable to sustain continuous contraction for long periods. Despite of this it has been considered as the responsible by the high pressure area, registered by manometry into the pharyngoesophageal transition. For this reason, it has been the object of therapeutics that promote the rupture of its integrity. AIMS: To give the anatomical bases to define the limits of participation of the cricopharyngeal muscle in the pharyngoesophageal transition function. To consider a morphological and functional alternative to explain the high pressure area on pharyngoesophageal transition and the implications of the myotomy, use of the botulinum toxin and balloon dilatation on pharyngoesophageal transition function. MATERIAL AND METHOD: Study of the laryngopharyngeal region in their morphologic characteristics and relationships on 24 pieces obtained from adults' corpses of both sexes fixed in 10% formaldehyde solution. RESULTS: The cricopharyngeal muscle presenting its anterior-lateral insertion, with a C-shaped outline, on the posterior-lateral edge of the cricoid cartilage. This kind of morphology blocks the possibility to generate a predominant anterior and posterior high pressure during its contraction like that we find at the pharyngoesophageal transition. The observation of this kind of pressure has its explanation in a tweezers-like relationship exerted on one side by the vertebral body and on the other side by the posterior contour of the cricoid cartilage. CONCLUSIONS: The muscular organization of the laryngopharyngeal segment allowed us to sustain that a large myotomy of the pharyngoesophageal transition, that takes more than just the cricopharyngeal transversal fasciculus, hinders the ejection function in a region where the dimension do not need any parietal sectioning. Myotomy that encompasses only the transversal fasciculus can contribute to improve the pharyngoesophageal flux by a decrease of the local resistance. The efficiency of this myotomy depends mostly on some residual pharyngeal ejection force and also on a slight hyolaryngeal displacement. The transversal fasciculus of the cricopharyngeal muscle is a narrow strip of muscular mass to be injected by percutaneous way with solution of botulinum toxin; maybe endoscopically. For this reason, dose, dilution and injection sites have an important meaning in the cricopharyngeal therapeutics using botulinum toxin. The efficiency of this procedure, like myotomy, depends on some residual pharyngeal ejection force and on, at least, some hyolaryngeal displacement. The dilation of the pharyngoesophageal transition with pneumatic balloon does not seem to be an adequate procedure for a region that does not present a narrow lumen determined by fibrosis. For anatomical characteristics of the TFE region, mean pressure as registered by the manometric method does not evaluate either the effectiveness or inadequacy of surgical myotomy, denervation or dilation using pneumatic balloon. PMID- 14762475 TI - [Pancreatic insulinomas: diagnosis and treatment]. AB - BACKGROUND: Despite its rarity, the insulinoma is the most common pancreatic neuroendocrine tumor. OBJECTIVE: Analyze clinical and immunohistochemical data from surgical resection of the pancreas insulinoma. METHOD: Twelve cases are described, concerning surgical aspects, complications and medium-long term outcome of patients. They underwent surgical treatment due to clinical suspicion and biochemical diagnosis of hypoglycemia and hyperinsulinism. RESULTS: The insulinoma was identified preoperatively in seven patients, while intraoperative ultrasonography and palpation were necessary for diagnosis in the other cases. Eight patients underwent pancreatic resection and pancreatic leak was observed in seven cases. Two patients developed diabetes mellitus and no mortality occurred in the current series. CONCLUSION: All patients presented satisfactory outcome and remained asymptomatic with normal glicemia levels. PMID- 14762476 TI - Transjugular liver biopsy: histological diagnosis success comparing the trucut to the modified aspiration Ross needle. AB - BACKGROUND: Transjugular liver biopsy is an alternative procedure for patients who present contraindications to standard percutaneous procedure. AIM: To compare the rate of histological diagnosis obtained on transjugular liver biopsy with an automated trucut needle and with a modified Ross needle. PATIENTS / METHOD: Eighty-five patients with suspicion of chronic liver diseases and presenting contraindications for percutaneous liver biopsy (coagulopathy, massive ascites, morbid obesity, or chronic renal problems) were submitted to 89 transjugular liver biopsies between March 1994 and April 2001 at "Hospital Sao Jose, Irmandade da Santa Casa de Misercordia", Porto Alegre, RS, Brazil. Thirty-five patients underwent 36 biopsies with an automated trucut needle, and 50 patients underwent 53 biopsies with a modified Ross needle. RESULTS: Histological diagnosis was reached in 32/35 subjects submitted to transjugular liver biopsy with the trucut needle (91%) and in 35/50 (70%) submitted to biopsy with the modified Ross needle. Specimens obtained with the trucut needle were significantly larger and less fragmented than those obtained with the Ross needle. CONCLUSION: Transjugular liver biopsy with the automated trucut needle allowed a higher rate of histological diagnosis when compared to the modified Ross needle in patients with suspicion of chronic liver diseases. PMID- 14762477 TI - [Nutritional status and dietary assessment of patients with gastrectomy]. AB - BACKGROUND: Nutrition is a crucial factor in gastric resection surgery and the most suitable alimentary canal reconstruction method must be considered in order to reduce the risk of malnutrition. The cause of postgastrectomy malnutrition has not been clearly determined, but the mechanisms behind malnutrition are evidently multifactorial. AIM: To evaluate the nutritional status of patients who underwent different reconstructive procedures after total or subtotal gastrectomy. PATIENTS AND METHODS: Fifty patients who have undergone gastrectomy for 0.5-39 years were assessed. The surgical procedures used were Billroth I in 7, Billroth II in 26, Henley in 3 and Roux-en-Y in 14 of the patients. Twenty one of them have followed gastrectomy for cancer. The nutritional status was evaluated by subjective global assessment, dietary recall and anthropometry. RESULTS: According to subjective global assessment, 6 of 50 patients were mild malnourished. The mean body mass index was 22 4.75 kg/m2, the average daily calorie intake was 1624 477 Kcal. Of the patients operated for cancer, those who underwent subtotal gastrectomy followed by Roux-en-Y presented higher body mass index. No relationship between the period of time since surgery with body mass index or with calorie intake was found. CONCLUSIONS: Among patients operated for cancer, subtotal gastrectomy with Roux-en-Y reconstruction is associated with better nutritional status. Factors other than low calorie intake are the cause of weight loss in patients with gastrectomy. PMID- 14762478 TI - [Extracolonic manifestations of familial adenomatous polyposis: incidence and impact on the disease outcome]. AB - BACKGROUND: Familial adenomatous polyposis is a hereditary disease with autossomic and dominant features, frequently associated to many extracolonic manifestations. AIM: To report extracolonic manifestations incidence and to analyze its impact on the disease's outcome. PATIENTS AND METHODS: Revision of patient charts treated from 1977 to 2001, relating associated extracolonic manifestations and its complications. RESULTS: Among 59 familial adenomatous polyposis patients, 23 (38,9%) presented some extracolonic manifestations at diagnosis or during follow-up. There were registered 37 different extracolonic manifestations (1.6 per patient). The most common manifestations were osteomas and congenital hypertrophy of the retinal pigment epithelium, diagnosed in 25% and 20% of the investigated patients, respectively. Other extracolonic manifestations were represented by upper digestive adenomas, epidermoid cysts, desmoid tumor (seven each), gastric cancer (three) and tireoid cancer (two). Desmoid-associated complications were reported in six patients, being intestinal obstruction in four and hidronephrosis in two; two deaths (28,5%) were registered. CONCLUSIONS: Extracolonic manifestations incidence is high (40%) and may affect disease's outcome and patient's quality of life. For these reasons, the research, prevention and adequate treatment of extracolonic manifestations turn into vital importance in familial adenomatous polyposis patients. PMID- 14762479 TI - [High resolution chromoendoscopy in the differential diagnosis of neoplastic and non-neoplastic polyps]. AB - BACKGROUND: Magnifying colonoscopy brought the possibility of precise histologic diagnosis of colorectal lesions through their surface appearance. Despite the high accuracy of magnifying colonoscopy it is a specialized and expensive equipment not available in most medical centers. Due to these reasons the use of conventional colonoscopy with chromoscopy has been raised because this produce can reproduce most of the information previously obtained by magnifying colonoscopy. AIM: To determine the role of high resolution colonoscopy and indigo carmine chromoscopy for differential diagnosis between neoplastic and non neoplastic colorectal lesions through measurements of accuracy, sensitivity, specificity, positive and negative predictive values. PATIENTS / METHODS: It was performed a prospective study. Seventy-four colorectal polyps were evaluated in 54 patients. A high resolution Olympus Exera CFQ 160L colonoscope was used. After the identification of the lesions, they were dyed with indigo carmine 0,2% and classified according to Kudo's classification by a single observer. After resection, the polyps were submitted to histopathological examination. RESULTS: The endoscopic findings were compared to histopathologic results. The accuracy of the method was 79,7%, sensibility of 88,8%, specificity of 55%, positive predictive value of 84,2% and a negative predictive value of 64,7%. CONCLUSION: We can conclude that we must be careful to apply high resolution colonoscopy and chromoscopy because adenomatous lesions can be misdiagnosed as non-neoplastic. PMID- 14762480 TI - [The effect of bacterial lipopolysaccharide on the gastric emptying of rats: a pretreatment evaluation using dexamethasone and methylene blue]. AB - BACKGROUND: The nitric oxide might be a putative mediator of the decrease in gastric emptying induced by bacterial lipopolysaccharide in rats. AIM: For that, we evaluated the effect of the pretreatment intravenous with dexamethasone and methylene blue in the retardation process of gastric emptying induced by intravenous application of lipopolysaccharide in rats. Dexamethasone has been shown to inhibit the induction of NOS II (induced NO-synthase) while the methylene blue, that blocks the soluble guanylyl cyclase, inhibits nitric oxide synthases and, in addition, inactivates nitric oxide directly. MATERIAL AND METHODS: Male Wistar rats, specific patogenic free, were used after a 24 hour fast and 1 hour-water withdrawn. The pretreatment was performed using dexamethasone solutions (3 and 6 mg/kg), methylene blue (2 mg/kg) or sterile vehicle. The treatment consisted in the application of lipopolysaccharide (50 mug/kg) or vehicle. The time period between the pretreatment and treatment was 10 minutes, excluding the study with dexamethasone 6 mg/kg that was 1 hour. The gastric emptying was evaluated 1 hour after the lipopolysaccharide application, except for two studies with dexamethasone 3 mg/kg in which the time periods were 2 and 8 hours. A saline solution containing phenol red was used as the test meal. The gastric emptying was determined by measuring gastric retention 10 minutes after the orogastric infusion of the test meal. RESULTS: The pretreatment with dexamethasone or methylene blue and treatment with vehicle did not have effect in the gastric emptying comparing to the control group. We found that pretreatment with dexamethasone in the studies for 1 hour and 2 hours did not interfere in the retardation of the gastric emptying produced by endotoxin. Nevertheless, in the eighth period study with this drug there was a significant reduction of gastric retention in the endotoxin-treated animals in relation to the unpretreated ones. Meanwhile, the pretreatment with the methylene blue completely blocked the action of endotoxin on the gastric emptying in rats. CONCLUSION: These results suggest a possible involvement of nitric oxide on the effect of lipopolysaccharide in rat gastric emptying. PMID- 14762481 TI - Magnifying endoscopy of the duodenum with dye scattering method in a case with celiac disease. AB - AIM: To know the more detailed findings of the small intestinal mucosa with the use of a magnifying endoscope and a vital dye, and the efficacy of the both tools. PATIENT AND METHODS: A 54-year old female patient with celiac disease. The duodenal mucosa downward as far as the descending portion was observed with a magnifying endoscope (Olympus GIF HM) before and after spraying the mucosa with 0.1% indigo carmine. RESULTS: The endoscopy clarified the atrophy and edema of each villus, and scattering of the dye revealed shorter villi with the relatively longer villi remaining in islands. CONCLUSION: The combination of magnifying endoscopy and the dye scattering method is useful for closer observation of the intestinal mucosa in celiac diseases. PMID- 14762482 TI - Familial gastric cancer. AB - BACKGROUND: Familial aggregation of gastric cancer has pointed out to a possible hereditary and genetic factor involved in the carcinogenesis of this disease. The diffuse type gastric cancer patients are frequently younger and the tumor has locally infiltrative growth pattern early in its development. Observation of families with frequent early onset gastric cancer has led to the identification of a novel gene implicated in gastric cancer susceptibility: CDH1/E-cadherin. Diffuse familiar gastric cancer is defined as any family presenting: two first degree relatives with diffuse gastric cancer, one of them with age under 50 years or at least 3 first-degree relatives irrespective age of onset. CASE REPORT: The family reported by us does not fit in any of the classification proposed. The precise identification of these families by clinical and molecular tools is of great importance. The case reported is an example of a family that probably is a form of hereditary gastric cancer not yet fully understood. CONCLUSION: Soon there will be new criteria, possibly including genetic and molecular characteristics. PMID- 14762483 TI - [Small intestine transplantation]. AB - BACKGROUND: Significant progress has been made in clinical small bowel transplantation over the last decade mainly due advances in biotechnology and new immunosuppressive regiments. This transplantation has now been indicated to treat special cases of intestinal failure. AIM: This review highlights recent developments in the area of small bowel transplantation. MATERIAL AND METHODS: Over 600 reports on clinical and experimental small bowel transplantation were reviewed. Aspects concerning research development, different immunosuppressive strategies, patient and graft monitoring, and improvements in surgical techniques are discussed. RESULTS: About 700 small bowel transplantation were performed in 55 transplant centers, 44% intestine-liver, 41% isolated intestinal graft and 15% multivisceral transplantation. Rejection and infection are the main limitation of this procedure. Actual 5 years post transplantation graft survival of the total international experience is 46% for isolated intestinal graft, 43% for combined intestine-liver and nearly 30% for multivisceral transplantation. Higher graft and patient survival are seen at the more experienced centers. In a series of 165 intestinal transplantation at University of Pittsburgh, PA, USA, actuarial patient survival was reported to be over 75% at one year, 54% at 5 years and 42% at 10 years. Over 90% patients from Pittsburgh program resume an unrestricted oral diet. CONCLUSION: Small bowel transplantation has advanced from an experimental strategy to a feasible alternative for patients with permanent intestinal failure. Further refinements in graft acceptance, immunosuppressive regiments, infection management and prophylaxis, surgical techniques as well as appropriated patient referral and selection are crucial to improve outcomes. PMID- 14762484 TI - [Comparison between idiopathic achalasia and achalasia caused by Chagas' disease: a review on the publications about the subject]. AB - BACKGROUND: Although idiopathic achalasia and achalasia caused by Chagas' disease have the same clinical manifestations and treatment, both with destruction of the esophageal myenteric plexus, it is possible that there are differences in the alterations of esophageal motility between the two diseases, caused by different grades of impairment of the excitatory and inhibitory esophageal neurons. AIMS: We performed a review of papers with results about the pathophysiology and esophageal motility alterations in idiopathic achalasia and Chagas' disease. DATE SOURCES: We reviewed papers which included data about the characteristics of idiopathic achalasia and Chagas' disease. DATA SYNTHESIS: Impairment of inhibitory esophageal neurons was shown in the two diseases. The results of the studies of the effects of atropine, edrophonium and botulin toxin suggested that the excitatory innervation is more intensely impaired in Chagas' disease than in idiopathic achalasia, explaining the increase in the lower esophageal sphincter pressure found in achalasia. The patients with Chagas' disease have more circulating muscarinic cholinergic receptor M2 autoantibodies than patient with idiopathic achalasia. The duration of the contractions in the esophageal body is longer in idiophatic achalasia than in Chagas' disease. CONCLUSIONS: The papers that studied Chagas' disease and idiopathic achalasia, mainly those which studied both diseases with the same methods, suggested that there are different grades of esophageal involvement by the two diseases, mainly the most important involvement of excitatory innervation in Chagas' disease than in idiopathic achalasia. PMID- 14762485 TI - The main indications and techniques for vascular exclusion of the liver. AB - BACKGROUND: The purpose of vascular clamping during the course of liver resection is to reduce bleeding and subsequent complications. AIM: To show both step-by step surgical techniques for vascular exclusion of the liver and their indications. METHODS: It is described the following techniques: clamping of the hepatic pedicle, "Pringle" maneuver; intermittent clamping of the hepatic pedicle; intermittent vascular exclusion of the liver, without vena cava clamping, and hepatic vascular exclusion with vena cava clamping. Also metabolic and homodynamic consequences as well as the technical failure of the application of each of them are discussed. CONCLUSIONS: The choice of technique to use for clamping during hepatectomy depends on the surgeon's judgment. Dogmatic or systematic attitude, is prejudiciable for the patient and liver surgeon must be able to use all kinds of clamping. PMID- 14762486 TI - Transplants: bioethics and justice. AB - Bioethics, as a branch of philosophy that focuses on questions relative to health and human life, is closely tied to the idea of justice and equality. As such, in understanding the concept of equality in its original sense, that is, in associating it to the idea to treat "unequals" (those who are unequal or different, in terms of conditions or circumstances) unequally (differentially), in proportion to their inequalities (differences), we see that the so-called "one and-only waiting list" for transplants established in law no. 9.434/97, ends up not addressing the concept of equality and justice, bearing upon bioethics, even when considering the objective criteria of precedence established in regulation no. 9.4347/98, Thus, the organizing of transplants on a one-and-only waiting list, with a few exceptions that are weakly applicable, without a case by case technical and grounded analysis, according to each particular necessity, ends up institutionalizing inequalities, condemning patients to happenstance and, consequently, departs from the ratio legis, which aims at seeking the greatest application of justice in regards to organ transplants. We conclude, therefore, that from an analysis of the legislation and of the principles of bioethics and justice, there is a need for the creation of a collegiate of medical experts, that, based on medical criteria and done in a well established manner, can analyze each case to be included on the waiting list, deferentially and according to the necessity; thus, precluding that people in special circumstances be treated equal to people in normal circumstances. PMID- 14762487 TI - Peri-intraventricular hemorrhage in newborns weighing less than 1500 grams: comparative analysis between 2 institutions. AB - PURPOSE: This study aims to characterize the peri-intraventricular hemorrhages in the neonatal period in very low birth weight newborns in 2 institutions that provide neonatal tertiary assistance. METHOD: This was a comparative and observational study in 2 neonatal intensive care units, the Maternity Hospital of Campinas and the "Centro de Aten o Integrada Sa de da Mulher" of the State University of Campinas, from December 01, 1998 to November 30, 1999. We examined 187 newborns for peri-intraventricular hemorrhages, using transfontanel ultrasound (76 and 11 respectively at the first and second unit), and classified them into 4 grades. We observed their gender, intrauterine growth, weight, and gestational age at birth. RESULTS: We diagnosed 34 cases of peri-intraventricular hemorrhages (13 and 21, respectively), and both groups differed as to the birth weight and the adequacy of weight to the gestational age at birth. There was no difference in the prevalence or extent of peri-intraventricular hemorrhages among cases. There was a statistically significant occurrence of lower birth weight at gestational ages of less than 30 weeks. CONCLUSIONS: The prevalence of peri intraventricular hemorrhages in our study was compared to that reported in the world literature. Although the cases of the second institution had a smaller mean birth weight, the prevalence of peri-intraventricular hemorrhages was similar to that at the first institution, probably because in the first one, 69% of the gestational ages of the neonates with hemorrhage were less than 30 weeks as compared to 48% in the second one. We stress the importance of the ultrasonographic method for diagnosing peri-intraventricular hemorrhages in very low birth weight newborns. PMID- 14762488 TI - Results of microsurgical anastomosis in men with seminal tract obstruction due to inguinal herniorrhaphy. AB - The incidence of vasal injury during inguinal herniorrhaphy is estimated at 0.5%. We sought to assess the patency rates and long-term fertility outcome after microsurgical repair of vasal obstruction related to prior inguinal herniorrhaphy. METHODS: Twenty procedures were performed on 13 men diagnosed with infertility and vasal injury secondary to previous inguinal herniorrhaphy. Eight of these men had undergone bilateral and 5 unilateral inguinal herniorrhaphy. Twelve procedures were vasovasostomies, 3 were crossover vasovasostomies, 2 were vasoepididymostomies, and 3 were crossover vasoepididymostomies. Eight patients were azoospermic, 2 were severely oligospermic (<1 M/mL), 1 was oligospermic, and 2 were asthenospermic. Patency data was obtained on all 13 patients, and pregnancy data was available for 10 couples (77%), with a mean follow-up of 69.5 months. RESULTS: The overall patency rate was 65%. In the vasovasostomy group, the patency rate was 60% (9/15), and in the vasoepididymostomy group it was 80% (4/5). Among the azoospermic patients, 13 procedures were performed. The patency rate was 42.9% for the vasovasostomy (3/7), and 100% for the vasoepididymostomy procedure (4/4). The overall pregnancy rate was 40%. Of the men who underwent vasoepididymostomy, 80% (4/5) established a pregnancy. CONCLUSIONS: Microsurgical vasovasostomy after inguinal vas injury results in a reasonable patency rate but a lower pregnancy rate than that after vasectomy reversal. When microsurgical vasoepididymostomy was possible, it resulted in high patency and pregnancy rate. Crossover vasoepididymostomy, when appropriate, can be a useful alternative to inguinal vasovasostomy. PMID- 14762489 TI - C-cells in colloid goiter. AB - PURPOSE: The aim of this investigation was to quantitatively evaluate C-cells in colloid goiters, analyzing 36 thyroids that were obtained through thyroidectomy from 24 patients with goiter and 12 normal glands from adult patients without thyroid disease, which were used as the control group. MATERIAL AND METHODS: On average, 6 different thyroid areas were sampled and labeled by immunohistochemistry with a monoclonal anticalcitonin antibody, utilizing the avidin-biotin-peroxidase complex. C-cells were counted in fields measuring 1 square centimeter, and the mean number of cells per field was then calculated. Data were statistically analyzed using the Mann-Whitney test. RESULTS: In the colloid goiter group, the number of C-cells ranged from 0 to 23 per field, while in normal controls they ranged from 20 to 148 per field. CONCLUSIONS: These results demonstrate a significant decrease of C-cell number in the colloid goiter group compared with control group, indicating that the hyperplastic process is restricted to follicular cells, to the detriment of C-cells, which probably cease to receive trophic stimuli. PMID- 14762490 TI - Prevalence of antinuclear autoantibodies in the serum of normal blood dornors. AB - OBJECTIVE: To examine the presence of serum antinuclear autoantibodies in a healthy population. METHODS: Serum of 500 normal blood donors between 18 and 60 years of age were tested for the presence of autoantibodies. Antinuclear antibodies were detected by indirect immunofluorescence technique using HEp-2 epithelial cells as the substrate. The presence of dnaN was detected by indirect immunofluorescence technique using Critidia lucillae as the substrate. Anti-SSA (RO), anti-SSB (LA), anti-Sm, and anti-RNP were determined by double radial immunodiffusion. RESULTS: In the evaluation of the presence of serum antibodies, antinuclear antibodies were detected in 22.6% of the sera. The presence of other antibodies was not significant. The majority of the titers were 1:40. CONCLUSION: The presence of autoantibodies is not necessarily pathologic and has to be related to the age group, gender, and clinical condition of the patient. PMID- 14762491 TI - Hemolytic disease of the newborn due to anti-U. AB - Anti-U is a rare red blood cell alloantibody that has been found exclusively in blacks. It can cause hemolytic disease of the newborn and hemolytic transfusion reactions. We describe the case of a female newborn presenting a strongly positive direct antiglobulin test due to an IgG antibody in cord blood. Anti-U was recovered from cord blood using acid eluate technique. Her mother presented positive screening of antibodies with anti-U identified at delivery. It was of IgG1 and IgG3 subclasses and showed a titer of 32. Monocyte monolayer assay showed moderate interaction of Fc receptors with maternal serum with a positive result (3.1%). The newborn was treated only with 48 hours of phototherapy for mild hemolytic disease. She recovered well and was discharged on the 4th day of life. We conclude that whenever an antibody against a high frequency erythrocyte antigen is identified in brown and black pregnant women, anti-U must be investigated. PMID- 14762492 TI - A critical review of the possible benefits associated with homeopathic medicine. AB - OBJECTIVE: To evaluate the recent scientific research progress on homeopathy. METHODOLOGY: Homeopathy was evaluated in terms of its clinical research; in vitro research, and physical foundations. The Medline database was the main reference source for the present research, concerning data of approximately the last 10 years. Secondary references (not available in this database) were obtained by means of direct requests to authors listed in the primary references. RESULTS: Clinical studies and in vitro research indicate the inefficacy of homeopathy. Some few studies with positive results are questionable because of problems with the quality and lack of appropriate experimental controls in these studies. The most recent meta-analyses on the topic yielded negative results. One of the few previous meta-analyses with positive results had serious publication bias problems, and its results were later substantially reconsidered by the main authors. The sparse in vitro homeopathic research with positive results has not been replicated by independent researchers, had serious methodological flaws, or when replicated, did not confirm the initial positive results. A plausible mechanism for homeopathic action is still nonexistent, and its formulation, by now, seems highly unlikely. CONCLUSIONS: As a result of the recent scientific research on homeopathy, it can be concluded that ample evidence exists to show that the homeopathic therapy is not scientifically justifiable. PMID- 14762493 TI - Ecstasy intoxication: the toxicological basis for treatment. AB - Youngsters are increasingly using 3,4 methylenedioxymethamphetamine, known as ecstasy, because it is wrongly believed that it does not induce harm. However, there are many reports of adverse effects, including acute intoxication, abuse potential, and possible neurotoxic effects. Therefore, health care providers need to promptly recognize the symptoms of systemic intoxication in order to initiate early treatment. The drug is used by the oral route for long hours during crowded dance parties. Acutely, ecstasy increases the release of serotonin and decreases its reuptake, leading to hypertension, hyperthermia, trismus, and vomiting. There is debate on whether recreational doses of ecstasy cause permanent damage to human serotonergic neurons. Ecstasy users showed a high risk of developing psychopathological disturbances. The prolonged use of ecstasy might induce dependence, characterized by tolerance and hangover. Acute ecstasy intoxication needs emergency-type treatment to avoid the dose-dependent increase in adverse reactions and in severity of complications. There are no specific antidotes to be used during acute intoxication. Supportive measures and medical treatment for each one of the complications should be implemented, keeping in mind that symptoms originate mainly from the central nervous system and the cardiovascular system. PMID- 14762494 TI - Letters to the editor. PMID- 14762495 TI - Editor'S note. PMID- 14762496 TI - A randomized, controlled, crossover trial of oral midazolam and hydroxyzine for pediatric dental sedation. AB - The effectiveness of oral midazolam in pediatric dentistry is controversial. This randomized, controlled, crossover, double blind clinical trial was conducted in order to study the effect of midazolam, used either alone or in association with hydroxyzine, during child dental treatment. Thirty seven dental sedation sessions were carried out on 11 ASA I uncooperative children less than five years-old. In each appointment children were randomly assigned to groups: P - placebo, M - midazolam (1.0 mg/kg), or MH - midazolam (0.75 mg/kg) plus hydroxyzine (2.0 mg/kg). Vital signs (blood pressure, breathing rate, pulse and oxygen saturation) and behavior parameters (consciousness, crying, movement, overall behavior) were evaluated every 15 minutes. Friedman and Wilcoxon statistical tests were used to compare groups and different moments in the same group. Normal values of vital signs were usually registered. Heart rate increased in groups P and M as the session went on. Group M presented less crying and movement at the first 15 minutes of treatment. Group MH caused more drowsiness at the beginning of the session. Overall behavior was better in group M than in groups P or MH. Group M produced effective sedation in 77% of the cases, and group MH did so in 30.8%. It was concluded that midazolam was effective and safe, and its association with hydroxyzine did not lead to additional advantages in pediatric dental sedation. PMID- 14762497 TI - Evaluation of the permeability of the furcation area of deciduous molars conditioned with Er:YAG laser and cyanoacrylate. AB - The purpose of this study was to evaluate in vitro the dentin permeability of the deciduous pulp chamber floor after employing 2-octyl cyanoacrylate and Er:YAG laser. Twenty four deciduous molars were used, divided into four groups. After chemical-surgical preparation each group received a different treatment: Group 1 control, without treatment; Group 2 - the floor of the pulp chamber was covered with a fine layer of 2-octyl cyanoacrylate; Group 3 - the floor of the pulp chamber was irradiated with Er:YAG laser (250 mJ, 10 Hz for 30 seconds, 80 J of energy and 320 pulses), and covered with a fine layer of 2-octyl cyanoacrylate; and Group 4 - the floor of the pulp chamber was irradiated with Er:YAG laser set at the parameters already described. After that the specimens received application of 0.5% methylene blue, for 15 minutes. The teeth were cut, photographed, and the digitalized images were analyzed using the ImageLab program. The results obtained were submitted to statistical analysis. Group 4 (Er:YAG) presented the largest averages in percentage of dye penetration area (19.5%), followed by Group 1 (11.1%), Group 3 (1.4%) and Group 2 (0.2%). The experimental model allowed to conclude that the specimens conditioned with 2 octyl cyanoacrylate (Group 2) and Er:YAG laser associated to 2-octyl cyanoacrylate (Group 3) presented a decrease in permeability, and the specimens treated with Er:YAG laser (Group 4) presented an increase in permeability of the analyzed area. PMID- 14762498 TI - Yeasts from the oral cavity of children with AIDS: exoenzyme production and antifungal resistance. AB - The oral fungal microbiota of 30 children with AIDS, of both genders, aged from two to six years, receiving outpatient treatment, was evaluated and compared with that of a control group composed of 30 healthy subjects with matching ages and genders. Virulence factors, such as exoenzyme production, and susceptibility to five antifungal agents using an E-Test kit were evaluated. C. albicans predominated over other species in the AIDS group, showing a higher production of proteinase and phospholipase when compared with that observed in the control group. In this study few clinical manifestations of and low selectivity for C. albicans (23.3%) were observed in the AIDS group. The enzymatic studies showed that 53.8% of the AIDS strains were strongly positive whereas only 33.3% of the non-AIDS strains were positive. Amphotericin B was the most effective drug among the antifungal agents tested against C. albicans. The frequency, selectivity and level of exoenzyme production by C. albicans suggest a higher pathogenicity in the AIDS children than in the control children. PMID- 14762499 TI - Fluorine content of several brands of chocolate bars and chocolate cookies found in Brazil. AB - Chocolate bars and chocolate cookies are foodstuffs highly appreciated by children. The possibility of having fluorine (F) among their components, associated with an excessive consumption, may make them decisive contributors to the total daily F intake. Thus, they could participate in the establishment of dental fluorosis. The aim of this study was to analyze the fluorine concentration [F] of the chocolates bars (CB) Baton, Confeti, Garoto Ball, Kinder Ovo, M&M s, Milkybar, Nescau, Nescau Ball, Surpresa, Surpresa Bichos, Tortuguita; and of the chocolate cookies (CC) Danyt s, Hipop , Nescau, Passatempo, Pokemon, S tio do Pica-Pau Amarelo and Trakinas. Samples were purchased in Bauru, Sao Paulo, Brazil. Three grams of each product were previously ashed at 525 C (CB and cookies fillings) and at 550 C (cookies dough), during 4 hours. Fluorine was separated from the ash by hexamethyldisiloxane (HMDS)-facilitated diffusion. Fluorine analysis was carried out with the specific electrode. Mean [F]s SD and amplitude (unit mg/g) were: CB = 0.30 0.45 (0.07 - 1.60, n = 12) and CC = 1.08 2.64 (0.04 - 7.10, n = 7). It was concluded that some of the analyzed foods may be important contributors to the total daily F intake. As for the product that had the highest [F] (Danyt s), when only 3 units are consumed just once a day, they may supply up to 40% of the maximum recommended daily F intake (0.07 mg/kg body weight) for a 2-year-old child (12 kg). The [F] in these products should be informed on their labels. PMID- 14762501 TI - Viability of autogenous bone grafts obtained by using bone collectors: histological and microbiological study. AB - The use of autogenous bone grafts is considered to be the best choice for reconstructive surgery. In the periodontal literature, the utilization of osseous coagulum was suggested by the end of the sixties. The purpose of this study is to consider the use of bone collectors (bone traps) as an alternative method for obtaining material to fill small bone imperfections, such as fenestrations and dehiscences. Thirty samples were obtained from bone drilling during fixture installation in patients (13 men and 17 women, with an average age of 54 years) requiring treatment at the Department of Periodontology and Implant Dentistry, University of Santo Amaro. These samples were fixed in 10% neutral formaldehyde for 24 hours and subjected to histological preparation, in order to evaluate the presence of viable osteoblasts. In addition, the material was placed in a fluid thioglycolate medium and incubated for 24 hours at 36 1 C in aerobiosis and anaerobiosis. Bacterial growth evaluation was made by using six different culture media (MacConkey agar, blood agar base, mannitol salt agar, Anaerokit LTD medium, Anaerokit LTD - bile medium, Anaerinsol). The results show that, if proper care is taken to prevent saliva contamination during the surgical procedure, this method of collecting autogenous bone may be useful in situations where small amounts of bone are required. PMID- 14762500 TI - Effect of a 0.5% chlorhexidine gel on dental plaque superinfecting microorganisms in mentally handicapped patients. AB - A randomized clinical trial was conducted to investigate the effect of a 0.5% chlorhexidine (CHX) gel on dental plaque superinfecting microorganisms in mentally handicapped patients. Thirty inmates from the institution "Casas Andre Luiz" were assigned to either test group (CHX gel, n = 15) or control group (placebo gel, n = 15). The gel was administered over a period of 8 weeks. Supragingival plaque samples were collected at baseline, after gel use (8 weeks) and 16 weeks after baseline. The presence of Gram-negative Enterobacteriaceae, Staphylococcus and yeasts was evaluated. No significant growth of any superinfecting microorganism was observed in the CHX group, when compared to the placebo group. The results indicated that the 0.5% chlorhexidine gel did not produce an undesirable shift in these bacterial populations. PMID- 14762502 TI - A modified pH-cycling model to evaluate fluoride effect on enamel demineralization. AB - Since in vitro pH-cycling models are widely used to study dental caries, they should allow evaluations of fluoride effect on early stages of caries development. Therefore, acid etching on enamel surface must be avoided, enabling surface microhardness (SMH) analysis. In the present study, the pH-cycling model originally described by Featherstone et al.9 (1986) was modified to preserve the enamel surface and to produce early carious lesions that could be evaluated using SMH and cross-sectional microhardness (CSMH) measurements. In order to validate this modified model, a dose-response evaluation with fluoride was made. Human enamel blocks with known SMH were submitted to such regimen with the following treatments: distilled deionized water (DDW; control) and solutions containing 70, 140 and 280 ppm F. Data from %SMH change and deltaZ (mineral loss) showed a statistically significant negative correlation between F concentration in treatment solutions and mineral loss. In conclusion, the modified pH-cycling model allowed the evaluation of changes on the outermost enamel layer during caries development, and a dose-response effect of fluoride reducing enamel demineralization was observed. PMID- 14762503 TI - Evaluation of the fluoride stability of dentifrices sold in Manaus, AM, Brazil. AB - The minimum requirement for the anti-caries effect of a dentifrice is the presence of available and stable fluoride in the formulation. The concentration of available fluoride in the major dentifrices sold in Brazil has been reported, but few data have been published about its stability, which is temperature dependent. Thus, the aim of this study was to evaluate the concentration and stability of fluoride in dentifrices sold in Manaus, AM, Brazil, which is a typical tropical city. The concentrations of total fluoride, total soluble fluoride, MFP, and of insoluble fluoride of six Brazilian dentifrices and an imported one were analyzed. The analyses were made when the dentifrices were purchased and during a year of storage at room temperature (28.9 1.16 C) and under refrigeration (26.3 0.88 C). The analyses were performed using an ion specific electrode Orion 96-09. At the time of purchase, all the dentifrices analyzed showed more than 1,000 ppm (mug F-/g; w/w) of soluble fluoride. However, in most of them, this form of fluoride was not shown to be stable. The highest loss of soluble fluoride was found for storage at room temperature, reaching up to 40%. Although all dentifrices comply with the Brazilian guidelines with regard to the concentration of total fluoride (maximum of 0.15%), the instability of soluble fluoride observed in some of them can impair their anti-caries effect and this condition is not contemplated in the Governmental rules. PMID- 14762504 TI - The influence of handling on the elasticity of addition silicone putties. AB - The handling of vinyl polysiloxane (addition silicone) impression putties with latex gloves is said to interfere with the setting of these impression materials. The aim of this study is to evaluate the effect of handling techniques on the setting of vinyl polysiloxane impression putties using several types of gloves. The setting of these materials was evaluated by means of an elasticimeter. Four vinyl polysiloxane putty impression materials and five brands of gloves (one made of vinyl, one of synthetic rubber, and three of natural rubber) were studied. Based on the type of glove, they were previously washed or not, and a spatula was used or not for initial mixing (before handmixing). The vinyl, the synthetic and one of the natural rubber gloves did not require the previous washing procedure and/or the use of a spatula for initial mixing. Two other natural rubber gloves - depending on the silicone -, showed satisfactory results only when the initial mixing was performed with a spatula. It was concluded that setting inhibition depends on the kind of vinyl polysiloxane impression material and the kind of gloves used, but when the initial mixing was performed with the spatula this setting inhibition was overcome. The results of this study also showed that it is possible to associate cross-contamination control and satisfactory performance of addition silicone putty materials. When doubts arise from the compatibility between vinyl polysiloxane impression putties and gloves, the initial mixing should be performed with a spatula. PMID- 14762505 TI - The effect of long-term storage on the microleakage of composite resin restorations: qualitative and quantitative evaluation. AB - The aim of this study was to evaluate the influence of storage periods of 24 hours and 3 months on the microleakage of class II cavities. Two methods of assessing microleakage were also compared. Class II cavities were prepared in sound human molars. MO cavities were restored using ABF experimental (Kuraray Medical Inc.) + Z250 composite resin (3M ESPE), and DO cavities were restored using Single Bond (3M ESPE) + Z250. After different storage periods, specimens were thermocycled, immersed in a dye (0.5% methylene blue solution for 4 h) and longitudinally sectioned. Dye penetration was scored according to a 0-4 scale. The extent of microleakage was measured using the ImageLab 2000 program. A statistically significant correlation was verified between both evaluation methods (r = 0.978, p < 0.001). ANOVA revealed a statistically significant difference between the tested adhesive systems regarding microleakage (p < 0.001), although it was not influenced by the different storage periods. PMID- 14762506 TI - Evaluation of the bending strength of rigid internal fixation with absorbable and metallic screws in mandibular ramus sagittal split osteotomy: in vitro study. AB - The aim of this study was to evaluate, in vitro, the bending strength of internal fixation with absorbable and metallic screws in mandibular ramus sagittal split osteotomy in sheep hemimandibles. The screws were inserted as lag screws, with an inverted "L" configuration, and the set was submitted to bending strength tests. The load and displacement of the peak and final load averages were, respectively, 18.45 kgf, 8.19 mm and 14.38 kgf for Group I, and 16.67 kgf, 6.73 mm and 13.98 kgf for Group II. The results were submitted to statistical analysis by Student s t test and by the Pearson correlation analysis. The groups showed no statistically significant differences, indicating the feasibility of both for osteosynthesis in mandibular ramus sagittal split osteotomies. PMID- 14762508 TI - In vitro evaluation of the effects of the interaction between irrigating solutions, intracanal medication and Er:YAG laser in dentin permeability of the endodontic system. AB - The purpose of this study was to evaluate in vitro the effects of different associations between irrigating solutions (EDTA-T and citric acid), intracanal medicament (NDP), and Er:YAG laser irradiation on dentin permeability. Fifty-one extracted single-rooted teeth were instrumented and divided into seven groups. Groups GI and GII had final irrigation with a demineralizing solution only (EDTA T and citric acid, respectively). Groups GIII and GIV had final irrigation with EDTA-T and citric acid, respectively, plus an association of irrigating solution and Er:YAG laser. Groups GV and GVI had final irrigation with EDTA-T and citric acid, respectively, plus an association of intracanal medication and Er:YAG laser. Group GVII (control group) had final irrigation with distilled water. All root canals were filled with NDP associated with rhodamine B dye. After the experimental period, the samples were transversely cut into six 2.0 mm thick slices for subsequent reading using the ImageLab software. Analysis of the results allowed us to conclude that there were statistically significant differences (p < 0.05) between the groups as to the penetration of the dye intracanal medication solution. Groups III and IV presented smaller values of dentinal permeability when compared to the other groups. The best results were obtained with the interaction between a demineralizing irrigating solution and the association of intracanal medicament and laser Er:YAG (groups V and VI). In these groups the observed penetration of the intracanal medicament plus dye solution in the apical third was, on average, 29% greater than in the other groups. PMID- 14762507 TI - Polymerization shrinkage evaluation of three packable composite resins using a gas pycnometer. AB - Modern restorative dentistry has been playing an outstanding role lately since composite resins, allied to adhesive systems, have been widely applied on anterior and posterior teeth restorations. The evolution of composite resins has mostly been verified due to the improvement of their aesthetic behavior and the increase in their compressive and abrasive strengths. In spite of these developments, the polymerization shrinkage inherent to the material has been a major deficiency that, so far, has been impossible to avoid. Using a gas pycnometry, this research investigated the polymerization shrinkage of three packable composite resins: Filtek P60 (3M), Prodigy Condensable (Kerr), and SureFil (Dentsply/Caulk), varying the distance from the light source to the surface of the resins (2 mm or 10 mm). The pycnometer Accupyc 1330 (Micromeritics, USA) precisely records helium displacement, allowing fast and reliable measurements of the volume of composite resin immediately before and after polymerization, without interference of temperature or humidity. Results were not found to be statistically different for the three tested resins, either for 2 mm or 10 mm-distance from the light source to the composite surface. PMID- 14762509 TI - Centric relation registration: intra- and interexaminer agreement after a calibration program. AB - Centric relation (CR) has been considered a maxillomandibular position of choice for some dental and prosthetic procedures. Although regarded as a fully reproducible relation, there is great controversy about its clinical use and recording technique, especially in patients with signs and symptoms of temporomandibular disorders (TMD). This study aimed at evaluating the effectiveness of a training program on intra- and interexaminer agreement when determining the clinical record of the CR position. Forty individuals constituted the sample, divided into symptomatic (TMD) and asymptomatic groups. Three previously calibrated examiners performed the initial assessment and the second evaluation after 30 days in a blind design, using Dawson s bilateral manipulation technique with and without an anterior stop. The amount of frontal and sagittal deviations in relation to intercuspal position, the presence of pain and discomfort during manipulation, and the first occlusal contact in CR were analyzed. Kendall and Kappa tests with a 5% level of significance were used to determine agreement. Values for both intra- and interexaminer agreement were deemed good. The best results were obtained for frontal deviations and assessment of pain (or absence of it) during manipulation. Sagittal deviations showed the lowest agreement in both examinations. The authors concluded that a calibration program could be effective for intra- and interexaminer agreement when recording centric relation. However, caution is recommended when analyzing some isolated items. PMID- 14762510 TI - Leishmanial infection: analysis of its first steps. A review. AB - The first steps in leishmaniasis are critical in determining the evolution of the disease. Major advances have recently been done in understanding this crucial moment. Fundamental research in parasite-vector interaction, parasite biology, insect saliva, and vertebrate host response have shed new light and uncovered a most fascinating and complex moment in leishmaniasis. We review here some of these aspects and we try to connect them in a logical framework. PMID- 14762511 TI - Prevalence of human papillomavirus types in women with pre-neoplastic and neoplastic cervical lesions in the Federal District of Brazil. AB - As a contribution to the public health authorities in planning prophylactic and therapeutic vaccine strategies, we describe the prevalence of human papillomavirus (HPV) types in women presenting abnormal cytological results in Pap smear screening tests in the Federal District, Central Brazil. We studied 129 cervical scraping samples from women whose cytological tests showed either pre neoplastic or neoplastic lesions. Amplification of HPV DNA was performed by polymerase chain reaction using consensus primers MY09 and MY11 followed by identification of isolates by restriction fragment length polymorphism. We detected HPV DNA in 62% of the samples, including HPV-16 in 43.8%, HPV-58 in 12.5%, HPV-31 in 10%, HPV-53 in 6.3%, each of HPV-18 and HPV-33 in 3.8% of the isolates. Other types (HPV-35, -52, -66, -CP8304, -6, -11, and -CP8061) were less frequent (= or < 2.5% each). The prevalence of HPV-58 was relatively higher in this population than in data in South America, but similar to results obtained in other studies in Latin America, Europe, and Eastern Asia. Case-control studies need to be carried out to establish the association between the prevalence of HPV types specially the less frequent high-risk types and cervical cancer. PMID- 14762512 TI - [Source of Chagas disease in Arcadia, state of Rio de Janeiro, Brazil]. AB - Following the report of triatomine nymphs in a house in Arcadia, Miguel Pereira, state of Rio de Janeiro, Brazil, the infested dwelling was checked. Several eggs and 46 specimens of Triatoma vitticeps (Stal, 1859) were collected. Among them, adults and nymphal instars accounted for 43.5% and 56.5%, respectively. Analysis of blood meals showed the ecletism of this species; 24 (52.2%) were single feeds, 18 insects (39.1%) fed on two hosts and 4 (8.7%) on three hosts. Trypanosoma cruzi infection rate of examined specimens was 13%. Finally one of the residents of the house was positive for anti-T. cruzi antibodies using indirect immunofluorescence. PMID- 14762513 TI - First report of Lymnaea columella Say, 1817 (Pulmonata: Lymnaeidae) naturally infected with Fasciola hepatica (Linnaeus,1758) (Trematoda: Digenea) in Argentina. AB - We report the first evidence of natural infection of Lymnaea columella with Fasciola hepatica in Argentina. A sample of 601 snails was collected in May 2003 in northeastern Corrientes, a province bounded on the north by Paraguay, on the east by Brazil and on the southeast by Uruguay. Among 500 examined snails, 44 (8.8%) were exclusively infected with F. hepatica. Parasite identification was based on morphological features of cercariae from snails, and of eggs and adult flukes from Wistar rats. We discuss the events suggesting that an enzootic transmission cycle of F. hepatica has been recently established in northeastern Corrientes. PMID- 14762514 TI - Angiostrongylus costaricensis and experimental infection of Sarasinula marginata II: elimination routes. AB - Angiostrongylus costaricensis intermediate hosts are terrestrial mollusks mostly belonging to the Veronicellidae family. In the present investigation we focused on the mechanisms of larval expulsion from Sarasinula marginata infected with A. costaricensis. Twenty-five mollusks were individually infected with 5000 L1 and sacrificed at 30 min and 1, 2, 4, 6, and 8 h post-infection and at days 1, 2, 4, 5, 6, 8, 10, 11, 12, 14, 15, 16, 20, 21, 22, 25, 26, 28, and 30 post-infection; the mollusks were then fixed and stained. Diverse organs involved throughout the course of the migratory routes of larvae from oral penetration on were specified and the mechanisms of larval access to the fibromuscular layer through the kidney, rectum, and vascular system were defined. The elimination of L3, derived from oral and/or cutaneous infections, appears to depend on granulomas located close to the excretory ducts of mucous cells. PMID- 14762515 TI - Feeding, defecation, and development times of Meccus longipennis Usinger, 1939 (Hemiptera: Reduviidae: Triatominae) under laboratory conditions. AB - Aspects related to hatching, time-lapse between presenting the blood-meal and beginning of feeding, feeding time, postfeed defecation delay, mortality, and fecundity for each stage of Meccus longipennis life-cycle were evaluated. The bugs were maintained in a dark incubator at 27 1 C and 80 5% rh, were fed weekly and checked daily for ecdysis or death. The hatching rate observed for 300 eggs was 76.7% and the average time of hatching was 19.8 days. Mean time-lapse between presentation of the blood meal and the beginning of feeding was under 5 min in nymphal stages and postfeed defecation delay was under 10 min in most stages, except in fourth and fifth stages. Mean feeding time was longer than 10 min in most stages, except in fourth stage. One hundred thirty-one nymphs (N) (65.5%) completed the cycle and the average time from NI to adult was 192.6 34.8 days. The average span in days for each stage was 18.1 for NI, 21.4 for NII, 29.5 for NIII, 45.5 for NIV and 55.9 for NV. The number of bloodmeals at each nymphal stage varied from 1 to 5. The mortality rate was 3.29 for NI, 6.8 for NII, 2.92 for NIII 3.76 for NIV, and 10.16 for NV nymphs. The average number of eggs laid per female in a 9-month period was 615.6. Based on our results, we conclude that M. longipennis has some biological and behavioral characteristics which influence its capacity of becoming infected and transmitting Trypanosoma cruzi to human populations in those areas of Mexico where it is currently present. PMID- 14762516 TI - Ultrastructural study of the in vitro interaction between Biomphalaria glabrata hemocytes and Schistosoma mansoni miracidia. AB - Biomphalaria glabrata and Schistosoma mansoni relationship was studied by light microscopy (LM) and freeze-fracture replica technique (FFR). We observed very thin cytoplasmic extensions of hemocytes in the LM, which then surround immobilize the miracidia. FFR images showed that the contact site between hemocytes cytoplasmic extensions and the external tegumentary coat involved only superficial layers of miracidia. Numerous vacuoles and filopodia were observed in the hemocyte cytoplasm, the latter binding with those from neighboring cells. In spite of the close interfilopodia contact, no cellular junctions were seen at these sites nor between filopodia-miracidia contact areas. The observed migration of hemocytes and their disposition in layers surrounding the miracidia in vitro correspond to previous studies. PMID- 14762517 TI - Flight metabolism in Panstrongylus megistus (Hemiptera: Reduviidae): the role of carbohydrates and lipids. AB - The metabolism of lipids and carbohydrates related to flight activity in Panstrongylus megistus was investigated. Insects were subjected to different times of flight under laboratory conditions and changes in total lipids, lipophorin density and carbohydrates were followed in the hemolymph. Lipids and glycogen were also assayed in fat body and flight muscle. In resting insects, hemolymph lipids averaged 3.4 mg/ml and significantly increased after 45 min of flight (8.8 mg/ml, P < 0.001). High-density lipophorin was the sole lipoprotein observed in resting animals. A second fraction with lower density corresponding to low-density lipophorin appeared in insects subjected to flight. Particles from both fractions showed significant differences in diacylglycerol content and size. In resting insects, carbohydrate levels averaged 0.52 mg/ml. They sharply declined more than twofold after 15 min of flight, being undetectable in hemolymph of insects flown for 45 min. Lipid and glycogen from fat body and flight muscle decreased significantly after 45 min of flight. Taken together, the results indicate that P. megistus uses carbohydrates during the initiation of the flight after which, switching fuel for flight from carbohydrates to lipids. PMID- 14762518 TI - An alternative technique to reveal polysaccharides in Toxoplasma gondii tissue cysts. AB - Ultrathin sections of tissue cysts isolated from the brain of Toxoplasma gondii infected mice were submitted to two different methodologies derived from the periodic acid - Schiff's reagent (PAS) technique. The use of osmium tetroxide vapor as a developing agent of the aldehyde oxidation to reveal polysaccharides with periodic acid resulted in positive reaction in amylopectin granules in bradyzoites, as well as in the wall and matrix of the cysts, with excellent increment of the ultrastructural morphology. This technique can be used for study of T. gondii-host cell intracellular cycle, the differentiation tachyzoite bradyzoite, and also for the formation of cysts into the host cells. PMID- 14762519 TI - Host nutritional status as a contributory factor to the remodeling of schistosomal hepatic fibrosis. AB - Weaning Swiss mice were percutaneously infected with 30 cercariae of Schistosoma mansoni and submitted to a shifting either from a deficient to a balanced diet or vice-versa, for 24 weeks. The nutritional status was weekly evaluated by measurements of growth curves and food intake. Hepatic fibrosis and periovular granulomas were studied by histological, morphometric and biochemical methods. All mice fed on a deficient diet failed to develop periportal "pipestem" fibrosis after chronic infection. An unexpected finding was the absence of pipestem fibrosis in mice on normal diet, probably related to the sample size. The lower values for nutritional parameters were mainly due to the deficient diet, rather than to infection. Liver/body weight ratio was higher in "early undernutrition" group, after shifting to the balanced diet. Volume density and numerical density of egg granulomas reached lowest values in undernourished animals. The amount of collagen was reduced in undernourished mice, attaining higher concentrations in well-fed controls and in "late undernutrition" (balanced diet shifted to a deficient one), where collagen deposition appeared increased in granulomas. That finding suggested interference with collagen degradation and resorption in "late" undernourished animals. Thus, host nutritional status plays a role in connective tissue changes of hepatic schistosomiasis in mice. PMID- 14762520 TI - Some coccidial parasites of the lizard Amphisbaena alba (Reptilia: Amphisbaenia: Amphisbaenidae). AB - Five parasites are described in the lizard Amphisbaena alba (Amphisbaenidae) from the state of Par , North Brazil. Mature oocysts of Choleoeimeria amphisbaenae n. sp., are passed already mature in the faeces. They are ellipsoidal-cylindrical, average 33.7 x 22.8 m and are devoid of micropyle, oocyst residuum or polar body. The colourless wall is smooth and of 2 layers. The 4 dizoic sporocysts have no Stieda body and average 13 x 9.3 m. Endogenous stages develop in the epithelial cells of the gall-bladder in the manner described for the genus and may cause extensive tissue damage. Sporulation of Isospora capanemaensis n. sp., is completed 3 days after the oocysts are voided in the faeces. They average 14.8 x 14.5 m and have no micropyle, oocyst residuum or polar body. The 2 tetrazoic sporocysts are pear-shaped, average 8.6 x 6.6 and have an inconspicuous Stieda body. Endogenous development is in the epithelial cells of the ileum, and heavy infections cause considerable tissue destruction. Multisporocystic oocysts passed in the faeces of one A. alba possibly originated from an invertebrate host ingested by the lizard. A globidium-like cyst in the digestive tract of A. alba measured 105 x 85 m and contained many hundreds of merozoites. A stained kidney smear of the same lizard revealed the presence of an unidentified parasite producing multinucleate cyst-like stages. PMID- 14762521 TI - Hemilucilia segmentaria (Fabricius, 1805) (Diptera: Calliphoridae) as new biological vector of eggs of Dermatobia hominis (Linnaeus Jr., 1781) (Diptera: Oestridae) in Reserva Biologica do Tingua, Rio de Janeiro, Brazil. AB - The aim of this note was to record for the first time the finding of Hemilucilia segmentaria acting as biological vector of Dermatobia hominis, during a study of the diversity of Calliphoridae at Reserva Biol gica do Tingua, Rio de Janeiro, Brazil. The insects were captured using traps baited with chicken vicera, for a period of 28-30 h twice per month. In the period of one year, 1987 insects were collected, 7.5% of which belonged to the H. segmentaria; of these a female was captured in May 2001, carrying a mass of 20 eggs on the left side of its abdomen. PMID- 14762522 TI - [Morphometric study of the genus Panstrongylus Berg, 1879 (Hemiptera, Reduviidae, Triatominae)]. AB - The subfamily Triatominae is actually represented by 137 species distributed among 6 tribes and 19 genera. Within this subfamily, the genus Panstrongylus, Berg 1879, is composed by 13 species widespread in sylvatic, peridomestic, and domestic habitats of Neotropical regions. These species are vectors of Chagas disease and consequently are found associated with its main hosts, such as birds and mammals. Interest in species of this genus has been increasing in the last few years. Reports of several authors indicate these Triatominae to invade and colonize houses, increasing their epidemiological significance. Morphometry was used in this study to investigate correlations among possible closely related species. We measured 224 specimens among 13 species through a set of metric variables of the head. The results indicated that the genus Panstrongylus seems to be homogeneous since 10 of the 14 species were shown to be closely related. PMID- 14762523 TI - Differential expression of adhesion moleculesshaping the T-cell subset prevalence during the early phase of autoimmune and Trypanosoma cruzi-elicited myocarditis. AB - The participation of cell adhesion molecules (CAMs) in the establishment of autoimmune and infectious myocarditis is an important matter of investigation and may have therapeutic implication. Trypanosoma cruzi infection induces a CD8 mediated myocarditis in patients with severe cardiomyopathy and experimental animals. Previously, we have proposed that this predominance of CD8+ T-cells is, at least in part, consequence of the differential expression of CAMs on circulating CD8+ lymphocytes. In the present study we investigated the participation of CAMs in shaping the phenotypic nature of the autoimmune CD4 mediated myosin-induced and the CD8-mediated T. cruzi-elicited myocarditis. We provide evidence that the prevalence of a certain T-cell subset inside the inflamed heart reflects the differential profile of the adhesion molecules VLA-4, LFA-1, and ICAM-1 displayed on a large proportion of this particular T-cell population in peripheral blood during the early phase of inflammation. Further, the expression of VCAM-1, ligand for VLA-4, and ICAM-1, counter-receptor for LFA 1, was up-regulated on vascular endothelium and paralleled the entrance of inflammatory cells into the cardiac tissue. Thus, this up-regulated expression of receptors-counter-receptors that regulate T-cell transmigration through the vascular endothelium may have an important role in the pathogenesis of the early phase of both autoimmune and infectious myocarditis. PMID- 14762524 TI - Parameters affecting cellular invasion and escape from the parasitophorous vacuole by different infective forms of Trypanosoma cruzi. AB - In this study we have examined certain aspects of the process of cell invasion and parasitophorous vacuole escape by metacyclic trypomastigotes and extracellular amastigote forms of Trypanosoma cruzi (G strain). Using Vero (and HeLa) cells as targets, we detected differences in the kinetics of vacuole escape by the two forms. Alcalinization of intercellular pH influenced both invasion as well as the escape from the parasitophorous vacuole by metacyclic trypomastigotes, but not the escape kinetics of extracellular amastigotes. We used sialic acid mutants as target cells and observed that the deficiency of this molecule facilitated the escape of both infective forms. Hemolysin activity was only detected in extracellular amastigotes and neither form presented detectable transialidase activity. Invasion of extracellular amastigotes and trypomastigotes in Vero cells was affected in different ways by drugs that interfere with host cell Ca2+ mobilization. These results are in line with previous results that indicate that metacyclic trypomastigotes and extracellular amastigote forms utilize mechanisms with particular features to invade host cells and to escape from their parasitophorous vacuoles. PMID- 14762525 TI - Antimicrobial activity in vitro of plumbagin isolated from Plumbago species. AB - Plumbagin is a naturally occurring naphthoquinone isolated from roots of Plumbago scandens. The plant was collected at the Campus of Fundacao Oswaldo Cruz, Rio de Janeiro, Brazil. P. scandens is used as a traditional medicine for the treatment of several diseases. The antimicrobial activity of plumbagin was evaluated using the macrodilution method. The compound exhibited relatively specific activity against bacteria and yeast. The minimum inhibitory concentration test showed the growth inhibiton of Staphylococcus aureus at a concentration of 1.56 g/ml and of Candida albicans at a concentration of 0.78 g/ml. These results suggest the naphthoquinone plumbagin as a promising antimicrobial agent. PMID- 14762526 TI - Antimicrobial activity of Hyptis ovalifolia towards dermatophytes. AB - The essential oil and the aqueous, hexane and methanolic fractions from Hyptis ovalifolia leaves were evaluated for their antifungal activity in vitro against 60 strains of dermatophytes: 10 strains of Microsporum canis, 10 of M. gypseum, 20 of Trichophyton rubrum and 20 of T. mentagrophytes. The extracts inhibited growth of the dermatophytes tested at different concentrations. The most biologically active was the essential oil from the leaves which inhibited 57 isolates (95%) at a concentration of 500 g/ml. PMID- 14762527 TI - Screening of Brazilian basidiomycetes for antimicrobial activity. AB - A total of 103 isolates of basidiomycetes, representing 84 species from different Brazilian ecosystems, were evaluated for their antifungal and antibacterial activity in a panel of pathogenic and non-pathogenic microorganisms. Tissue plugs of the fruiting bodies were cultivated in liquid media and the whole culture extracted with ethyl acetate. Crude extracts from Agaricus cf. nigrecentulus, Agrocybe perfecta, Climacodon pulcherrimus, Gloeoporus thelephoroides, Hexagonia hydnoides, Irpex lacteus, Leucoagaricus cf. cinereus, Marasmius cf. bellus, Marasmius sp., Nothopanus hygrophanus, Oudemansiella canarii, Pycnoporus sanguineus, Phellinus sp., and Tyromyces duracinus presented significant activity against one or more of the target microorganisms. Eight isolates were active only against bacteria while three inhibited exclusively the growth of fungi. Two extracts presented wide antimicrobial spectrum and were active against both fungi and bacteria. Differences in the bioactivity of extracts obtained from isolates from the same species were observed. PMID- 14762528 TI - Comparative evaluation of pyrethroid insecticide formulations against Triatoma infestans (Klug): residual efficacy on four substrates. AB - We investigated the residual efficacy of four insecticide formulations used in Chagas disease vector control campaigns: cyfluthrin 12.5% suspension concentrace (SC), lambda-cyhalothrin 10% wettable powder (WP), deltamethrin 2.5% SC, and 2.5% WP on four types of circular blocks of wood, straw with mud, straw with mud painted with lime, and mud containing 5% of cement. Three concentrations of these insecticides were tested: the LC90 (previously determined on filter paper), the double of the LC90, and the recommended operational dose. For each bioassay test, 15 third-stage nymphs of Triatoma infestans (Klug) (Hemiptera: Reduviidae) were exposed for 120 h to each treatment at 24 h, 30, 60, 90, and 180 days post spraying. Mortality rates, moulting history and behaviour were recorded at 24, 48, 72, and 120 h of exposure. Mortality rates were highest during the first 30 days post-spraying. Highest mortality rates (above 50%) were observed for deltamethrin 2.5% SC and lambda-cyhalothrin 10% WP on wood blocks up to three months post-spraying. Mud was the substrate on which treatments showed lowest persistence, with the other two substrates showing intermediate residual efficacy of all treatments. During the first 30 days WP formulations were not as effective as SC flowable formulations but, overall in the longer term, WP gave grater mortality rates of T. infestans nymphs exposed at up to six months post-spraying. Porous surfaces, especially mud, showed most variability presumably due to absorption of the insecticide. In contrast the less porous surfaces (i.e. wood and lime-coated mud) kept mortality rates high for longer post-treatment, irrespective of the insecticide concentration used. PMID- 14762529 TI - Activity of Euphorbia splendens var. hislopii N.E.B. (Euphorbiaceae) latex against Lymnaea columella (Say, 1817) (Pulmonata: Lymnaeidae), intermediate host of Fasciola hepatica, Linnaeus, 1758 (Trematoda: Fasciolidae). 2: limited field testing. AB - The molluscicidal evaluation of Euphorbia splendens var. hislopii (Crown of thorns) against Lymnaea columella snails, intermediate host of Fasciola hepatica, in irrigation ditches of the Pisciculture Station at Universidade Federal Rural do Rio de Janeiro, was studied under limited field conditions. An aqueous solution of the latex at 5 mg/l was tested in two irrigation ditches (experimental and control ditches), after initial sampling of the snail population present. Twenty-four hours after application of the product, it was verified that 97.4% of free L. columella snails and 100% of snails of the same species captive in cages and used as sentinels at three points equidistant from the application site in the experimental ditch, died. For Biomphalaria tenagophila and Melanoides tuberculata snails, present in the experimental ditch, the mortality was 100%, for the species Pomacea spp. the mortality was 40%. No mortality was verified in the free mollusks, or in the sentinels in the ditch used as control. E. splendens var. hislopii latex is thus an efficient natural molluscicide, which may be used as an alternative control agent against L. columella. PMID- 14762530 TI - [Not Available]. PMID- 14762531 TI - [A molecular public health?]. AB - This article focuses on the relationship between Public Health and Molecular Genetics. Both the notion of Public Health and correlate expressions are discussed in an attempt to establish their respective objects of investigation and fields of practice. In addition, a brief description is given of the precarious health conditions in Brazil. The author remarks on the limited social effectiveness of the role ascribed to public health professionals in Brazil. Techniques and concepts developed by Molecular Genetics are presented and their importance for Public Health is analyzed. Genetic risk is discussed and compared to the idea of inherited disposition to disease, with special emphasis on ethical and epistemological points of view. The notion of expert is considered in terms of its adequacy for training health professionals to deal with issues pertaining to Molecular Biology and Human Genetics in Public Health matters. Lastly, the author argues for the role of the state in defining social priorities in health. PMID- 14762541 TI - [Women's opinions on circumstances under which hospitals should perform abortions]. AB - Little is known about the opinion of Brazilian women on induced abortion. One objective of a study carried out in the region of Campinas, Sao Paulo State, was to learn whether women agreed as to specific circumstances under which hospitals should provide abortions. A total of 1838 women of childbearing age and who had been pregnant at least once were interviewed. Single women and those who had had at least one induced abortion were the ones who most agreed with hospitals performing abortions under the circumstances presented. The circumstances most accepted were rape, the woman's life being at risk, and fetal malformation. A smaller percentage of women were in favor of abortion when the reasons affected the woman but could not be observed in a direct and objective way by others. PMID- 14762542 TI - [Action-oriented participant research as a strategy for training and evaluation in health education]. AB - In this article we present the results of a study on educational practices at the "Escuela de Malariologia y Saneamiento Ambiental 'Dr. Arnoldo Gabaldon'" (EMSA), a pioneering institution in training health personnel, pertaining to the Ministry of Health and Social Assistance in Venezuela. This study was developed as an experiment in action-oriented participant research, i. e. authorities, teachers, and students were committed to an evaluation of the educational practices in which they were involved. The main results of this cooperative inquiry were: a theoretical reconstruction of the models of health education employed by EMSA; a critical analysis of those models; and the design and testing of an alternative model centered on community participation. PMID- 14762543 TI - [Reliability of data from Authorization Forms for Hospital Admittance, Rio de Janeiro, Brazil]. AB - This article presents data from a study on quality of information from the data bank on the form used in the Brazilian health care system to authorize hospital admittance (AIH). The form pertains to the Unified Health System (SUS) and is used by hospitals to be reimbursed for the health care provided to patients, with public funds. The AIH data bank is the largest source of information in Brazil on production by hospitals. This study analyzes a sample of forms filled out by private hospitals under contract with the city of Rio de Janeiro in 1986. Study design was based on reliability between interviewers, and agreement was analyzed utilizing the kappa test. Agreement was analyzed between data from AIH forms filled out by administrative personnel and information from medical files for patients, focusing on demographic, administrative, and clinical variables. The first two types of variables were generally more reliable than the third. Reliability for the principal diagnosis was worse than for the prescribed procedure, despite the latter being the reference for unit reimbursement by the Unified Health System. Furthermore, where there was disagreement, hospitals were more likely to adopt a procedure with higher reimbursement value as compared to the data derived from the clinical file. The article proposes measures to improve data quality. PMID- 14762544 TI - [Indicators of quality for medical records and ambulatory care in Maringa (state of Parana, Brazil), 1991: an exercise in evaluation]. AB - This study sought to establish health-care quality indicators based on data from routine clinical records produced by the Municipal Health Department in Maringa, Parana State. Records for 7813 patients seen in the month of October 1991 at three Integrated Health Centers (IHC) were assessed to determine whether the recorded information allowed for the formulation of quality indicators. Indicators of data quality, demand, coverage, resolubility, and geographic distribution of the clientele by IHC were formulated. Some 49.7% of records had no diagnostic hypothesis, and 14.8% of recorded diagnoses were illegible, 78% of visits were by residents of the IHC's cachement areas; there was a low estimated coverage of pre-natal care of 37%; an estimated 70% of children under one year of age residing in the area were seen during the month; patients seen in October 1991 had a mean of 3 physician visits per person during the preceding year and 4.3% of records specified a referral to another service. Patient records in the centers studied are precarious as information services for either routine internal use or external evaluation. In spite of these deficiencies, it was possible to extract some indicators of quality; however, significant improvements in recording are needed to support more in-depth evaluation. PMID- 14762545 TI - [Generic drugs: an alternative for the Brazilian market]. AB - The analysis of the main characteristics of the pharmaceutical industry worldwide and market aspects of that industry in Brazil leads us to consider a high share of external dependency and oligopolization. This situation is not peculiar to our country, but is rather a global trend. Recent Government initiatives in Brazil, aiming at reducing that dependency, associated with the critical aspects regarding the prices and costs of drugs as compared to international prices, are the background that support the proposal of evaluating generic drugs as an alternative for Brazilian market. One of the most striking aspect is the verification of overpricing of raw materials as demonstrated by "transfer prices", a practice that has been observed in several countries. Analysing international concepts dealing with generic drugs, generic names and pharmaceutical equivalence, based on World Health Organization recommendations and evaluating the experiences of different countries, several actions are proposed in order to gradually enforce a policy of generic drugs, bioequivalent and interchangeable, as a possible regulatory mechanisms for Brazil. PMID- 14762546 TI - [Occurrence of Vibrio parahaemolyticus in mussels (Perna perna, Linnaeus, 1758) from a natural coastal bed in the municipality of Palhoca, Santa Catarina, Brazil]. AB - A study was carried out on the occurrence of Vibrio parahaemolyticus in forty samples of mussels (Perna perna, Linnaeus, 1758) from a natural bed at Pinheira Beach, Municipality of Palhoca, Santa Catarina (Brazil), during a three-month period. The most probable number technique was used for isolation methodology with prior enrichment of samples in alkaline peptone water and subsequent planting on thiosulfate citrate bile salts sucrose agar. Vibrio parahaemolyticus was found in 52.5% of samples of mussels with a most probable number range of <3 to 93 organisms/g. Serotyping was performed on 61 isolates and our findings indicate that 36.1% of these isolates were serologically non-typable; 54.1% of isolates displayed only flagellate antigenic structures and 8.2% had both antigenic structures. None of the isolates were Kanagawa positive. PMID- 14762547 TI - [Economic development, inequality, and health]. AB - The author refers to the social, economic, individual, and collective dimensions of the relationship between health and disease. He considers that individuals and populations are not equally exposed to protective and risk factors. He highlights the uneven development between nations and Brazilian regions, and communities, as well as discussing the relationship between development, wealth, health, and social justice. The article analyzes development theories and makes a distinction between economic development and growth. The concepts of development trap and social refugees are coined. The author defines a healthy community, points to the need for a shift in the current development model, and delineates the health sector's role and limitations in dealing with social inequality. PMID- 14762549 TI - [Not Available]. PMID- 14762548 TI - [Nutrition and development]. AB - Discussions on development and nutrition frequently have a double focus: one that considers nutritional status as a reflection of development, and another that admits that nutrition strongly influences the level and rhythm of development. The authors view nutrition as a decisive factor for adequate social and economic development, specifying the fact that nutritional interventions might contribute to significant improvements in individual classroom performance, productivity at the workplace, and resistance to infections. The implications of malnutrition for the economy are thus evident: either by limitations in income expectations, increased costs deriving from burdens assumed by the population to protect themselves, or considerable hidden social costs--absenteeism, work accidents, and rotation of employees. Given modernization and a competitive economy, there is an unquestionable need for nutrition to head the agenda of national priorities, thus promoting human resource development and the preservation of this human capital, one of the chief goals of a socially responsible economy. PMID- 14762555 TI - [Not Available]. PMID- 14762556 TI - Sustaining community living of the very old: medical and service issues. AB - The unprecedented prolongation of life in the United States has been accompanied by increased numbers of disabled people in their 80s and 90s, who have high needs for health care and social services. The paper reports longitudinal findings on 150 individuals, 85 years and older. Over three years, their functioning on activities of daily living significantly declined at the same time that they continued to described their health as good and to report contentment about their life. Families were active in sustaining community living of the oldest old, but those most at risk of institutionalization were the childless. Over time, 48% of the survivors stayed functionally fit, while 28% became increasingly disabled and dependent and 24% remained stable at a high level of disability. The means by which the oldest old coped with their disabilities include practical steps in simplifying their environment and narrowing their social world. They also used cognitive regulation by modifying their health beliefs and delimiting those areas over which they could exert a sense of control. PMID- 14762557 TI - [Smoking habits among students at the Pelotas Medical School: prevalence in 1986 and 1991]. AB - A cross-sectional survey of smoking habits was carried out among students at the School of Medicine in Pelotas, State of Rio Grande do Sul, Brazil, in 1986 and 1991. The overall participation rate for the originally selected sample was 96% and 98% respectively. In 1986, regular smokers comprised 21.6% of the students and 14.6% in 1991; former smokers were 7.3% and 10.9%, respectively. There was no association between sex, parental smoking, and smoking habits of the students. Students in their last two years of medical training showed an increased prevalence in smoking habits (1991). Respiratory symptoms were associated with smoking habits. It was concluded that the prevalence of smoking is decreasing among students of the School of Medicine in Pelotas. PMID- 14762558 TI - [Conceptual aspects of educational practices for HIV/AIDS prevention]. AB - The importance and complexity of educational work related to HIV prevention demand a continuing discussion of its contents. Regarding the ideologic features, it must be assumed that, with the AIDS epidemic, medical science reaches a dimension of the life relatively preserved from its previous hegemonic position: sexual behavior. In this field, a coercive attitude on the part of health professionals may lead to negative reactions. To avoid this, individual and community participation and self-responsibility are central tools for the improvement of the efficacy of education practices. The author proposes three components that educational planners should consider: information and its effects on knowledge, attitudes, and perception related to HIV infection; response facilitation; and support for individual and collective reflection. PMID- 14762559 TI - [Representations, attitudes, and practices related to Cutaneous Leishmaniasis in people from Acosta County, San Jose province, Costa Rica. An exploratory anthropological study]. AB - An exploratory anthropological study on representations, attitudes, and practices related to Cutaneous Leishmaniasis (CL) among the rural population of Acosta County, Costa Rica, aimed to estimate the applicability of epidemiologically based control measures. Open interviews with a small sample of individuals from both case and control households provided the basic for a Propositional Discourse Analysis (PDA). Results are that Acosta people consider CL a distinct nosologic entity, but they are mainly interested in its clinical manifestations in children (who are mainly affected), as well as in their own capacity to act on the disease using folk remedies. The idea of control measures on reservoirs, on vectors, or on the spatial and temporal context of contact does not arise spontaneously in people's thinking. Nevertheless, CL is perceived as a disruption in the safe domiciliary and peridomiciliary space, so that control measures intervening there could have a chance for success. PMID- 14762560 TI - [Autonomy and work in medicine]. AB - We discuss autonomy in medical work as resulting from two basic moments: as social work consuming and producing goods and services, reproducing the dominant mode of production and its ethical foundations; and as a service in which autonomy, limited as social work, expresses some attributes acquired by medical work (technical independence and self-regulation) as a result of the monopoly in the use of science to deal with individually perceived health-related needs. These moments are expressed in the doctor-patient relationship, which makes uncertainties surrounding that relationship more evident. From this understanding about medical work and its autonomous condition, we discuss some theoretical concepts related to the study of labor and/or services in contemporary society and which provide us with an epistemological basis for more adequate approaches to scientific research concerning that object. PMID- 14762561 TI - The bioecology of snail vectors for schitosomiasis in Brazil. AB - This paper describes the ecology of two species of Biomphalaria: B. glabrata and B. straminea. These species have been intensively studied in Brazil and in other countries since the 1950s. The literature in this area can be broadly subdivided into three categories, linked to three distinct historical periods: 1) an accumulated store of early studies carried out in the laboratory and in the field; 2) the development of quantitative ecological models; and 3) the development of an alternative methodology for breeding snails under seminatural conditions. PMID- 14762562 TI - [Noise-induced hearing loss and its association with cumulative working time among urban bus workers]. AB - Through regression analysis the relationship between noise-induced hearing loss and variables "total cumulative working time", "arterial blood pressure" and "age" was examined. The study was carried out among 278 bus drivers and ticket takers, users of an occupational health center in Campinas, Sao Paulo State (Brazil). The main results were a positive association between hearing loss and cumulative working time, as well as an interaction between this variable and the bus workers' age. PMID- 14762563 TI - [Mosquito control based on larvicides in the State of Rio Grande do Sul, Brazil: choice of the control agent]. AB - A comparison between chemical and biological larvicides in routine operations against mosquitoes in Rio Grande do Sul State was carried out in this study. In laboratory bioassays against Culex quinquefasciatus, biological formulations Vectobac 12 AS and Teknar 3000 (Bacillus thuringiensis israelensis) as well as ABG 6262 (B. sphaericus 2362), both in liquid and powder form, were highly effective. Locally produced B. thuringiensis israelensis, formulations also yielded good results. Among chemical larvicides, pyrethroid compounds Pirisa and K-Othrine yielded better results than the organophosphates Lebaycid and Abate. These last formulations yielded responses ten weaker than predicted in other studies. Under field conditions, a dose of 1250 mg/m2 for biological formulations was considered adequate for routine application because at this level it is possible to overcome physical influences on results. Only B. sphaericus preparations caused important disruption of mosquito colonization in active breeding sites. Up to 39 weeks were tabulated without complete colonization in natural conditions and one month in artificially colonized tanks. This study suggests that biological alternatives should be considered in mosquito control programs. They may be a solution to such problems as resistance to larvicides, elimination of natural enemies, and short-lasting effects of applications. PMID- 14762564 TI - [Nutritional strategies for controlling iron deficiency anemia]. AB - This review was elaborated in order to contribute to the planning of strategies for controlling iron deficiency anemia in developing countries. The need for intervention should be determined by the degree of iron deficiency in the individual group and knowledge of its effects on quality of life, morbidity, and mortality. The most frequent approach is to provide iron supplementation during pregnancy, lactation, and early childhood as a basic primary healthcare measure. Fortification and dietary modification are complementary approaches, and should be developed. PMID- 14762565 TI - Inequality, violence, and ecology in Brazil. AB - This paper studies the articulation of three subjects--ecology, inequality, and violence--as interconnected components of reality, in such a way that each of the three interferes in the whole. At a time of increased ecological awareness, this paper shows that poverty, destination, and violence not only affect the environment, but that they are also responsible for the destruction of culture and life in society. Furthermore, as a cry for Nature, this increased awareness represents a struggle for life with dignity for all. PMID- 14762566 TI - [Brazilian risk map: limited applicability of a worker model]. AB - This paper shows the limitations identified in Brazilian risk map applications from a legal and methodological perspective. First, the conception, origin, and value of the risk-map methodology are presented as perceived under the Italian Health Reform. Next, the authors report on introduction of the methodology into Brazil in the early 1980s and the reactions by companies when it became a legal requirement for those which had Accident Prevention Committees. Finally, a brief discussion on the effectiveness of its use by workers in preventing health risks is included. PMID- 14762567 TI - Environmental and health education for school-age children: a transdisciplinary approach. AB - A brief critical analysis and suggestion for guidelines concerning environmental and health education in the elementary school are presented from the viewpoint of emerging pedagogic experiences and theoretical philosophical reflections. The major points discussed are: the importance of transdisciplinarity, the enhancement of participatory education where technical knowledge and popular wisdom meet, the planning and execution of concrete experiences which stimulate the sensations of pleasure and of admiration for nature and life, the analysis and the search for a solution to problems affecting the local reality of pupils without losing sight of global issues. A major priority is investment in teacher training, stimulating acquisition of knowledge through creative practice and formation of a critical awareness, essential for the school to clearly show its commitment to a future of greater social equality and harmony in our relationship to the environment. PMID- 14762568 TI - [Social movements and health: notes for a debate]. AB - This text is a contribution to the debate about the participation of social scientists in the field of health. The difficulties and perspectives involved in the incorporation of a specific theme such as social movements are treated. When criticizing the heavy emphasis traditionally given to "politics" in Brazilian social analysis, in reality attention is being called to the problem of the decrease in reflective power on a theoretically and politically relevant issue, the constitution of collective subjects in a specific period of time. This article represents an effort at overcoming the dilemma. PMID- 14762569 TI - Glutathione peroxidase induction protects Saccharomyces cerevisiae sod1deltasod2delta double mutants against oxidative damage. AB - Saccharomyces cerevisiae mutants deficient in superoxide dismutase genes (sod1delta, sod2delta and the double mutant) were subjected to H2O2 stress in the stationary phase. The highest sensitivity was observed in the sod2delta mutant, while the sod1deltasod2delta double mutant was not sensitive. Sod mutants had lower catalase activity (44%) than wild-type cells, independent of H2O2 stress. Untreated cells of sod1deltasod2delta double mutants showed increased glutathione peroxidase activity (126%), while sod1delta had lower activity (77%) than the wild type. Glutathione levels in sod1delta were increased (200-260%) after exposure to various H2O2 concentrations. In addition, the highest malondialdehyde levels could be observed without H2O2 treatment in sod1delta (167%) and sod2delta (225%) mutants. In contrast, the level of malondialdehyde in the sod1deltasod2delta double mutant was indistinguishable from that of the wild type. These results suggest that resistance to H2O2 by sod1deltasod2delta cells depends on the induction of glutathione peroxidase and is independent of catalase, and that glutathione is a primary antioxidant in the defense against H2O2 in stationary phase sod1delta mutants. PMID- 14762571 TI - Mathematical modeling of electro-rotation spectra of small particles in liquid solutions: application to human erythrocyte aggregates. AB - Electro-rotation can be used to determine the dielectric properties of cells, as well as to observe dynamic changes in both dielectric and morphological properties. Suspended biological cells and particles respond to alternating-field polarization by moving, deforming or rotating. While in linearly polarized alternating fields the particles are oriented along their axis of highest polarizability, in circularly polarized fields the axis of lowest polarizability aligns perpendicular to the plane of field rotation. Ellipsoidal models for cells are frequently applied, which include, beside sphere-shaped cells, also the limiting cases of rods and disks. Human erythrocyte cells, due to their particular shape, hardly resemble an ellipsoid. The additional effect of rouleaux formation with different numbers of aggregations suggests a model of circular cylinders of variable length. In the present study, the induced dipole moment of short cylinders was calculated and applied to rouleaux of human erythrocytes, which move freely in a suspending conductive medium under the effect of a rotating external field. Electro-rotation torque spectra are calculated for such aggregations of different length. Both the maximum rotation speeds and the peak frequencies of the torque are found to depend clearly on the size of the rouleaux. While the rotation speed grows with rouleaux length, the field frequency nu(p) is lowest for the largest cell aggregations where the torque shows a maximum. PMID- 14762570 TI - Heterofucans from Dictyota menstrualis have anticoagulant activity. AB - Fucan is a term used to denote a family of sulfated L-fucose-rich polysaccharides which are present in the extracellular matrix of brown seaweed and in the egg jelly coat of sea urchins. Plant fucans have several biological activities, including anticoagulant and antithrombotic, related to the structural and chemical composition of polysaccharides. We have extracted sulfated polysaccharides from the brown seaweed Dictyota menstrualis by proteolytic digestion, followed by separation into 5 fractions by sequential acetone precipitation. Gel electrophoresis using 0.05 M 1,3-diaminopropane-acetate buffer, pH 9.0, stained with 0.1% toluidine blue, showed the presence of sulfated polysaccharides in all fractions. The chemical analyses demonstrated that all fractions are composed mainly of fucose, xylose, galactose, uronic acid, and sulfate. The anticoagulant activity of these heterofucans was determined by activated partial thromboplastin time (APTT) using citrate normal human plasma. Only the fucans F1.0v and F1.5v showed anticoagulant activity. To prolong the coagulation time to double the baseline value in the APTT, the required concentration of fucan F1.0v (20 g/ml) was only 4.88-fold higher than that of the low molecular weight heparin Clexane (4.1 g/ml), whereas 80 g/ml fucan 1.5 was needed to obtain the same effect. For both fucans this effect was abolished by desulfation. These polymers are composed of fucose, xylose, uronic acid, galactose, and sulfate at molar ratios of 1.0:0.8:0.7:0.8:0.4 and 1.0:0.3:0.4:1.5:1.3, respectively. This is the fist report indicating the presence of a heterofucan with higher anticoagulant activity from brown seaweed. PMID- 14762572 TI - Lipid peroxidation and total radical-trapping potential of the lungs of rats submitted to chronic and sub-chronic stress. AB - Exposure to stress induces a cluster of physiological and behavioral changes in an effort to maintain the homeostasis of the organism. Long-term exposure to stress, however, has detrimental effects on several cell functions such as the impairment of antioxidant defenses leading to oxidative damage. Oxidative stress is a central feature of many diseases. The lungs are particularly susceptible to lesions by free radicals and pulmonary antioxidant defenses are extensively distributed and include both enzymatic and non-enzymatic systems. The aim of the present study was to determine lipid peroxidation and total radical-trapping potential (TRAP) changes in lungs of rats submitted to different models of chronic stress. Adult male Wistar rats weighing 180-230 g were submitted to different stressors (variable stress, N = 7) or repeated restraint stress for 15 (N = 10) or 40 days (N = 6) and compared to control groups (N = 10 each). Lipid peroxidation levels were assessed by thiobarbituric acid reactive substances (TBARS), and TRAP was measured by the decrease in luminescence using the 2-2'-azo bis(2-amidinopropane)-luminol system. Chronic variable stress induced a 51% increase in oxidative stress in lungs (control group: 0.037 +/- 0.002; variable stress: 0.056 +/- 0.007, P < 0.01). No difference in TBARS was observed after chronic restraint stress, but a significant 57% increase in TRAP was presented by the group repeatedly restrained for 15 days (control group: 2.48 +/- 0.42; stressed: 3.65 +/- 0.16, P < 0.05). We conclude that different stressors induce different effects on the oxidative status of the organism. PMID- 14762573 TI - Prolactin induces adrenal hypertrophy. AB - Although adrenocorticotropic hormone is generally considered to play a major role in the regulation of adrenal glucocorticoid secretion, several reports have suggested that other pituitary hormones (e.g., prolactin) also play a significant role in the regulation of adrenal function. The aim of the present study was to measure the adrenocortical cell area and to determine the effects of the transition from the prepubertal to the postpubertal period on the hyperprolactinemic state induced by domperidone (4.0 mg kg-1 day-1, sc). In hyperprolactinemic adult and young rats, the adrenals were heavier, as determined at necropsy, than in the respective controls: adults (30 days: 0.16 +/- 0.008 and 0.11 +/- 0.007; 46 days: 0.17 +/- 0.006 and 0.12 +/- 0.008, and 61 days: 0.17 +/- 0.008 and 0.10 +/- 0.004 mg for treated and control animals, respectively; P < 0.05), and young rats (30 days: 0.19 +/- 0.003 and 0.16 +/- 0.007, and 60 days: 0.16 +/- 0.006 and 0.13 +/- 0.009 mg; P < 0.05). We selected randomly a circular area in which we counted the nuclei of adrenocortical cells. The area of zona fasciculata cells was increased in hyperprolactinemic adult and young rats compared to controls: adults: (61 days: 524.90 +/- 47.85 and 244.84 +/- 9.03 microm2 for treated and control animals, respectively; P < 0.05), and young rats: (15 days: 462.30 +/- 16.24 and 414.28 +/- 18.19; 60 days: 640.51 +/- 12.91 and 480.24 +/- 22.79 microm2 ; P < 0.05). Based on these data we conclude that the increase in adrenal weight observed in the hyperprolactinemic animals may be due to prolactin-induced adrenocortical cell hypertrophy. PMID- 14762574 TI - Immunological effects of donor lymphocyte infusion in patients with chronic myelogenous leukemia relapsing after bone marrow transplantation. AB - Allogeneic bone marrow transplantation (alloBMT) is the only curative therapy for chronic myelogenous leukemia (CML). This success is explained by the delivery of high doses of antineoplastic agents followed by the rescue of marrow function and the induction of graft-versus-leukemia reaction mediated by allogeneic lymphocytes against host tumor cells. This reaction can also be induced by donor lymphocyte infusion (DLI) producing remission in most patients with CML who relapse after alloBMT. The immunological mechanisms involved in DLI therapy are poorly understood. We studied five CML patients in the chronic phase, who received DLI after relapsing from an HLA-identical BMT. Using flow cytometry we evaluated cellular activation and apoptosis, NK cytotoxicity, lymphocytes producing cytokines (IL-2, IL-4 and IFN-gamma), and unstimulated (in vivo) lymphocyte proliferation. In three CML patients who achieved hematological and/or cytogenetic remission after DLI we observed an increase of the percent of activation markers on T and NK cells (CD3/DR, CD3/CD25 and CD56/DR), of lymphocytes producing IL-2 and IFN-gamma, of NK activity, and of in vivo lymphocyte proliferation. These changes were not observed consistently in two of the five patients who did not achieve complete remission with DLI. The percent of apoptotic markers (Fas, FasL and Bcl-2) on lymphocytes and CD34-positive cells did not change after DLI throughout the different study periods. Taken together, these preliminary results suggest that the therapeutic effect of DLI in the chronic phase of CML is mediated by classic cytotoxic and proliferative events involving T and NK cells but not by the Fas pathway of apoptosis. PMID- 14762575 TI - Cytotoxicity of chlorhexidine digluconate to murine macrophages and its effect on hydrogen peroxide and nitric oxide induction. AB - Chlorhexidine, even at low concentrations, is toxic for a variety of eukaryotic cells; however, its effects on host immune cells are not well known. We evaluated in vitro chlorhexidine-induced cytotoxicity and its effects on reactive oxygen/nitrogen intermediate induction by murine peritoneal macrophages. Thioglycollate-induced cells were obtained from Swiss mice by peritoneal lavage with 5 ml of 10 mM phosphate-buffered saline, washed twice and resuspended (10(6) cells/ml) in appropriate medium for each test. Cell preparations contained more than 95% macrophages. The cytotoxicity was determined by the 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide assay and the presence of hydrogen peroxide (H2O2) and nitric oxide (NO) by the horseradish peroxidase dependent oxidation of phenol red and Griess reaction, respectively. The midpoint cytotoxicity values for 1- and 24-h exposures were 61.12 +/- 2.46 and 21.22 +/- 2.44 microg/ml, respectively. Chlorhexidine did not induce synthesis or liberation of reactive oxygen/nitrogen intermediates. When macrophages were treated with various sub-toxic doses for 1 h (1, 5, 10, and 20 microg/ml) and 24 h (0.5, 1, and 5 microg/ml) and stimulated with 200 nM phorbol myristate acetate (PMA) solution, the H2O2 production was not altered; however, the NO production induced by 10 microg/ml lipopolysaccharide (LPS) solution varied from 14.47 +/- 1.46 to 22.35 +/- 1.94 micromol/l and 13.50 +/- 1.42 to 20.44 +/- 1.40 micromol/l (N = 5). The results showed that chlorhexidine has no immunostimulating activity and sub-toxic concentrations did not affect the response of macrophages to the soluble stimulus PMA but can interfere with the receptor-dependent stimulus LPS. PMID- 14762576 TI - Production and characterization of monoclonal antibodies to a Brazilian bovine herpesvirus type 5. AB - Antigens of a bovine herpesvirus type 5 (BHV-5), isolated from a cow with a neurological infection in Rio Grande do Sul State, Brazil, were used to immunize BALB/c mice to produce monoclonal antibodies (mAbs). Eleven hybridomas secreting mAbs directed at BHV-5 antigens were obtained after two fusions and screening of 356 hypoxanthine-aminopterin-thymidine-resistant clones. The mAbs reacted at dilutions up to 1:500 (hybridoma culture supernatant) and up to >1:10,000 (ascitic fluid) in an indirect fluorescent antibody assay (IFA) and in immunoperoxidase staining of BHV-5-infected cells. Four mAbs (1D12, 2E2, 2G10 and 4E4) showed virus-neutralizing activity against the parental BHV-5 isolate. Five mAbs (1F3, 2A6, 2F9, 2G10 and HB24L) reacted in Western immunoblotting with a protein of approximately 90 kDa. Three other mAbs (2E2, 3D6 and 4E4) reacted in IFA with antigens of a BHV-1 mutant glycoprotein C- negative strain, demonstrating that they are directed at a viral antigen other than glycoprotein C. The eleven mAbs tested reacted with 20 BHV-5 field isolates and nine mAbs reacted with 10 BHV-1 isolates. Two mAbs (1F3 and 2F9) failed to react with BHV-1 field isolates, although they displayed a weak and nonreproducible reaction with the BHV-1 reference strain Los Angeles. These mAbs may be very useful in distinguishing between BHV-1 and BHV-5 infections since most of the traditional reagents and techniques are unable to do so. One mAb (2F9) was shown to bind to viral antigens by immunohistochemistry of histological sections of the brain of a BHV-5-infected calf. These results demonstrate that the mAbs produced here are suitable for use in a variety of immunological techniques and therefore may be useful for diagnostic and research purposes. PMID- 14762577 TI - Comparison of different monoclonal antibodies against immunosuppressive proteins of Ascaris suum. AB - The extract of Ascaris suum suppresses the humoral and cellular immune responses to unrelated antigens in the mouse. In order to further characterize the suppressive components of A. suum, we produced specific monoclonal antibodies which can provide an important tool for the identification of these proteins. The A. suum immunosuppressive fractions isolated by gel filtration from an extract of adult worms were used to immunize BALB/c mice. Popliteal lymph node cells taken from the immunized animals were fused with SP2/O myeloma cells and the cloned hybrid cells obtained were screened to determine the specificity of secreted antibodies. Three monoclonal antibodies named MAIP-1, MAIP-2 and MAIP-3 were selected and were shown to react with different epitopes of high molecular weight proteins from the A. suum extract. All antibody molecules have kappa-type light chains but differ in heavy chain isotype. MAIP-1 is a mouse IgM, MAIP-2 is an IgA immunoglobulin and MAIP-3 is an IgG1 immunoglobulin and they recognize the antigen with affinity constants of 1.3 x 10(10) M-1, 7.1 x 10(9) M-1 and 3.8 x 10(7) M-1, respectively. The proteins recognized by these monoclonal antibodies (PAS-1, PAS-2 and PAS-3) were purified from the crude extract by affinity chromatography and injected with ovalbumin in BALB/c mice in order to determine their suppressive activity on heterologous antibody production. It was demonstrated that these three proteins are able to significantly suppress anti ovalbumin antibody secretion, with PAS-1 being more efficient than the others. PMID- 14762578 TI - Contrast sensitivity to angular frequency gratings is not higher than to Cartesian gratings. AB - When contrast sensitivity functions to Cartesian and angular gratings were compared in previous studies the peak sensitivity to angular stimuli was reported to be 0.21 log units higher. In experiments carried out to repeat this result, we used the same two-alternative forced-choice paradigm, but improved experimental control and precision by increasing contrast resolution from 8 to 12 bits, increasing the screen refresh rate from 30 Hz interlaced to 85 Hz non-interlaced, linearizing the voltage-luminance relation, modulating luminance in frequencies that minimize pixel aliasing, and improving control of the subject's exposure to the stimuli. The contrast sensitivity functions to Cartesian and angular gratings were similar in form and peak sensitivity (2.4 cycles per visual degree (c/deg) and 32 c/360 degrees, respectively) to those reported in a previous study (3 c/deg and 32 c/360 degrees, respectively), but peak sensitivity to angular stimuli was 0.13 log units lower than that to Cartesian stimuli. When the experiment was repeated, this time simulating the experimental control level used in the previous study, no difference between the peak sensitivity to Cartesian and angular stimuli was found. This result agrees with most current models that assume Cartesian filtering at the first visual processing stage. The discrepancy in the results is explained in part by differences in the degree of experimental control. PMID- 14762579 TI - Measurement of Ki-67 antigen in 159 pituitary adenomas using the MIB-1 monoclonal antibody. AB - Pituitary adenomas sometimes show rapid growth and recurrence, and about one third invade the structures surrounding the sella turcica. In an attempt to determine aggressive behavior at an early stage, we used the MIB-1 antibody to identify the Ki-67 antigen. The present study was designed to evaluate pituitary adenomatous tissue in terms of secretion and proliferation and to correlate the Ki-67 index with hormone phenotype and invasive behavior. Material from 159 patients submitted to one or more resections of pituitary adenomas was evaluated. Forty-two non-secretory adenomas and 43 adenomas immunoreactive for growth hormone, 19 for prolactin, 18 for growth hormone and prolactin, 16 for adrenocorticotropic hormone (ACTH), and 21 cases of plurihormonal/gonadotropin adenomas were detected by immunohistochemistry. The MIB-1 antibody was positive in 139 samples and the Ki-67 index ranged from 0.16 to 15.48% (mean = 1.22 +/- 2.09%), with no significant difference between genders, age groups, or secretory and non-secretory status. The Ki-67 index was higher in ACTH-secreting adenomas. Invasive pituitary adenomas had a significantly higher Ki-67 index (2.01 +/- 3.15%) than macroadenomas with or without supra-sellar extension (1.12 +/- 1.87%; P = 0.02). The index was not significantly different in the subgroup of adenomas with invasion of the cavernous sinus compared to groups with other types of invasion. We conclude that tumoral proliferative activity evaluated by the detection of the Ki-67 antigen is significantly higher in invasive than noninvasive adenomas, information which can be useful in therapeutic postoperative management since index cut-off values associated with aggressive behavior can be established. PMID- 14762580 TI - The compulsive-like aspect of the head dipping emission in rats with chronic electrolytic lesion in the area of the median raphe nucleus. AB - Head dipping (HD) is a behavioral pattern considered to have a risk assessment or an exploratory role and is used as a complementary parameter to evaluate anxiety in experimental animals. Since rats with electrolytic lesion in the area of the median raphe nucleus displayed high frequencies of HD in a previous study, the present investigation was undertaken to confirm this observation and to determine its anxiety-related origin. HD episodes were counted in adult male Wistar rats (270-350 g) with electrolytic lesion (N = 11) and sham-lesioned controls (N = 12). When HD was measured for 60 min on an elevated open platform, lesioned rats emitted 13 times more HD than controls (264.7 +/- 93.3 vs 20.3 +/- 7.6 episodes), with the difference being statistically significant (P < 0.05). HD counts during 10-min sessions held 7, 14, 21, 27, and 63 days after lesion showed significantly higher means (range: 28.14 +/- 5.38 to 62.85 +/- 9.48) compared to sham-lesioned controls (range: 7.37 +/- 1.13 to 8.5 +/- 1.45). Normal rats stepped down into their home cages when the vertical distance between them and the cage was short (16 cm), and the step-down latencies increased with increasing depths (36.7 +/- 7.92 to 185.87 +/- 35.44 s). Lesioned rats showed a similar behavior when facing the shortest depth, but had a significantly increased number (23.28 +/- 2.35 episodes) and latency (300 +/- 0.00 s) of HD compared to normal rats (9.25 +/- 1.37 episodes and 185.87 +/- 35.44 s) when facing the greatest depth (30 cm). This suggests that HD may be a depth-measuring behavior related to risk assessment. PMID- 14762581 TI - Clinical features of panic patients sensitive to hyperventilation or breath holding methods for inducing panic attacks. AB - Our aim was to compare the clinical features of panic disorder (PD) patients sensitive to hyperventilation or breath-holding methods of inducing panic attacks. Eighty-five PD patients were submitted to both a hyperventilation challenge test and a breath-holding test. They were asked to hyperventilate (30 breaths/min) for 4 min and a week later to hold their breath for as long as possible, four times with a 2-min interval. Anxiety scales were applied before and after the tests. We selected the patients who responded with a panic attack to just one of the tests, i.e., those who had a panic attack after hyperventilating (HPA, N = 24, 16 females, 8 males, mean age +/- SD = 38.5 +/- 12.7 years) and those who had a panic attack after breath holding (BHPA, N = 20, 11 females, 9 males, mean age +/- SD = 42.1 +/- 10.6 years). Both groups had similar (chi(2) = 1.28, d.f. = 1, P = 0.672) respiratory symptoms (fear of dying, chest/pain discomfort, shortness of breath, paresthesias, and feelings of choking) during a panic attack. The criteria of Briggs et al. [British Journal of Psychiatry, 1993; 163: 201-209] for respiratory PD subtype were fulfilled by 18 (75.0%) HPA patients and by 14 (70.0%) BHPA patients. The HPA group had a later onset of the disease compared to BHPA patients (37.9 +/- 11.0 vs 21.3 +/- 12.9 years old, Mann-Whitney, P < 0.001), and had a higher family prevalence of PD (70.8 vs 25.0%, chi(2) = 19.65, d.f. = 1, P = 0.041). Our data suggest that these two groups--HPA and BHPA patients--may be specific subtypes of PD. PMID- 14762582 TI - Ionic radiocontrast inhibits endothelium-dependent vasodilation of the canine renal artery in vitro: possible mechanism of renal failure following contrast medium infusion. AB - To determine if radiocontrast impairs vascular relaxation of the renal artery, segments (4-5 mm in length) of canine renal artery were suspended in vitro in organ chambers to measure isometric force (95% O2/5% CO2, at 37 C). Arterial segments with and without endothelium were placed at the optimal point of their length-tension relation and incubated with 10 microM indomethacin to prevent synthesis of endogenous prostanoids. The presence of nonionic radiocontrast (iohexol, Omnipaque 350, 1 ml in 25 ml control solution, 4% (v/v)) did not alter endothelium-dependent relaxation to acetylcholine in rings precontracted with both norepinephrine and prostaglandin F2alpha (N = 6). When the rings were precontracted with prostaglandin F2alpha, the presence of ionic contrast did not inhibit the relaxation of the arteries. However, in canine renal arteries contracted with norepinephrine, the presence of ionic radiocontrast (diatrizoate meglumine and diatrizoate sodium, MD-76, 1 ml in 25 ml control solution, 4% (v/v)) inhibited relaxation in response to acetylcholine, sodium nitroprusside (N = 6 in each group), and isoproterenol (N = 5; P < 0.05). Rings were relaxed less than 50% of norepinephrine contraction. Following removal of the contrast, vascular relaxation in response to the agonists returned to normal. These results indicate that ionic radiocontrast nonspecifically inhibits vasodilation (both cAMP-mediated and cGMP-mediated) of canine renal arteries contracted with norepinephrine. This reversible impairment of vasodilation could inhibit normal renal perfusion and act as a mechanism of renal failure following radiocontrast infusion. In the adopted experimental protocol the isoproterenol-induced relaxation of renal arteries precontracted with norepinephrine was more affected, suggesting a pivotal role of the cAMP system. PMID- 14762583 TI - Protection of plasmid DNA by a Ginkgo biloba extract from the effects of stannous chloride and the action on the labeling of blood elements with technetium-99m. AB - Ginkgo biloba extract (EGb) is a phytotherapeutic agent used for the treatment of ischemic and neurological disorders. Because the action of this important extract is not fully known, assays using different biological systems need to be performed. Red blood cells (RBC) are labeled with technetium-99m (Tc-99m) and used in nuclear medicine. The labeling depends on a reducing agent, usually stannous chloride (SnCl2). We assessed the effect of different concentrations of EGb on the labeling of blood constituents with Tc-99m, as sodium pertechnetate (3.7 MBq), and on the mobility of a plasmid DNA treated with SnCl2 (1.2 microg/ml) at room temperature. Blood was incubated with EGb before the addition of SnCl2 and Tc-99m. Plasma (P) and RBC were separated and precipitated with trichloroacetic acid, and soluble (SF-P and SF-RBC) and insoluble (IF-P and IF RBC) fractions were isolated. The plasmid was incubated with Egb, SnCl2 or EGb plus SnCl2 and agarose gel electrophoresis was performed. The gel was stained with ethidium bromide and the DNA bands were visualized by fluorescence in an ultraviolet transilluminator system. EGb decreased the labeling of RBC, IF-P and IF-RBC. The supercoiled form of the plasmid was modified by treatment with SnCl2 and protected by 40 mg/ml EGb. The effect of EGb on the tested systems may be due to its chelating action with the stannous ions and/or pertechnetate or to the capability to generate reactive oxygen species that could oxidize the stannous ion. PMID- 14762584 TI - Photophysical studies of zinc phthalocyanine and chloroaluminum phthalocyanine incorporated into liposomes in the presence of additives. AB - The photophysical properties of zinc phthalocyanine (ZnPC) and chloroaluminum phthalocyanine (AlPHCl) incorporated into liposomes of dimyristoyl phosphatidylcholine in the presence and absence of additives such as cholesterol or cardiolipin were studied by time-resolved fluorescence, laser flash photolysis and steady-state techniques. The absorbance of the drugs changed linearly with drug concentration, at least up to 5.0 M in homogeneous and heterogeneous media, indicating that aggregation did not occur in these media within this concentration range. The incorporation of the drugs into liposomes increases the dimerization constant by one order of magnitude (for ZnPC, 3.6 x 10(4) to 1.0 x 10(5) M-1 and for AlPHCl, 3.7 x 10(4) to 1.5 x 10(5) M-1), but this feature dose does not rule out the use of this carrier, since the incorporation of these hydrophobic drugs into liposomes permits their systemic administration. Probe location in biological membranes and predominant positions of the phthalocyanines in liposomes were inferred on the basis of their fluorescence and triplet state properties. Both phthalocyanines are preferentially distributed in the internal regions of the liposome bilayer. The additives affect the distribution of these drugs within the liposomes, a fact that controls their delivery when both are used in a biological medium, retarding their release. The addition of the additives to the liposomes increases the internalization of phthalocyanines. The interaction of the drugs with a plasma protein, bovine serum albumin, was examined quantitatively by the fluorescence technique. The results show that when the drugs were incorporated into small unilamellar liposomes, the association with albumin was enhanced when compared with organic media, a fact that should increase the selectivity of tumor targeting by these phthalocyanines (for ZnPC, 0.71 x 10(6) to 1.30 x 10(7) M-1 and for AlPHCl, 4.86 x 10(7) to 3.10 x 10(8) M 1). PMID- 14762585 TI - Chronic experimental myocardial infarction produces antinatriuresis by a renal nerve-dependent mechanism. AB - The present study focused on the role of sympathetic renal nerve activity, in mediating congestive heart failure-induced sodium retention following experimental chronic myocardial infarction. Groups of male Wistar rats (240-260 g) were studied: sham-operated coronary ligation (CON3W, N = 11), coronary ligation and sham-operated renal denervation (INF3W, N = 19), 3 weeks of coronary ligation and sympathetic renal nerve denervation (INF3WDX, N = 6), sham-operated coronary ligation (N = 7), and 16 weeks of coronary ligation (INF16W, N = 7). An acute experimental protocol was used in which the volume overload (VO; 5% of body weight) was applied for 30 min after the equilibration period of continuous iv infusion of saline. Compared to control levels, VO produced an increase (P < 0.01, ANOVA) in urine flow rate (UFR; 570%) and urinary sodium excretion (USE; 1117%) in CON3W. VO induced a smaller increase (P < 0.01) in USE (684%) in INF3W. A similar response was also observed in INF16W. In INF3WDX, VO produced an immediate and large increase (P < 0.01) in UFR (547%) and USE (1211%). Similarly, in INF3W VO increased (P < 0.01) UFR (394%) and USE (894%). Compared with INF3W, VO induced a higher (P < 0.01) USE in INF3WDX, whose values were similar to those for CON3W. These results suggest that renal sympathetic activity may be involved in sodium retention induced by congestive heart failure. This premise is supported by the observation that in bilaterally renal denervated INF3WDX rats myocardial infarction was unable to reduce volume expansion-induced natriuresis. However, the mechanism involved in urinary volume regulation seems to be insensitive to the factors that alter natriuresis. PMID- 14762586 TI - Intermittent diazepam and continuous phenobarbital to treat recurrence of febrile seizures: a systematic review with meta-analysis. AB - Convulsions triggered by fever are the most common type of seizures in childhood, and 20% to 30% of them have recurrence. The prophylactic treatment is still controversial, so we performed a systematic review to find out the effectiveness of continuous phenobarbital and intermittent diazepam compared to placebo for febrile seizure recurrence. METHOD: Only randomized, double-blind, placebo controlled trials were analyzed. The recurrence of febrile seizure was assessed for each drug. RESULTS: Ten eligible clinical trials were included. Febrile seizure recurrence was smaller in children treated with diazepam or phenobarbital than in placebo group. Prophylaxis with either phenobarbital or diazepam reduces recurrences of febrile seizures. The studies were clinical, methodological, and statistically heterogeneous. CONCLUSION: The effectiveness of phenobarbital and diazepam could not be demonstrated because clinical trials were heterogeneous, and the recommendation for treatment recurrence should rely upon the experience of the assistant physician yet. PMID- 14762587 TI - Influence of intracranial hemorrhage and neonatal seizures on the neurological and psychomotor development of premature infants at Hospital de Clinicas de Porto Alegre, Brazil. AB - This study compared the results of neurodevelopmental examination at 6 months' corrected age of premature infants with neonatal seizures and/or intracranial hemorrhage and normal premature infants. There was a statistically significant correlation (p=0.000007) between intracranial hemorrhage and seizures in the group of 68 premature infants seen in the neurodevelopmental outpatient service at Hospital de Clinicas de Porto Alegre, Brazil. Intracranial hemorrhage was significantly associated with multiparity (p=0.02). The neurodevelopmental examination at 6 months' corrected age revealed that patients who suffered neonatal intracranial hemorrhage and/or seizures had inappropriate muscle tone, strength and reflexes, as well as delay in head control. CONCLUSION: We compared the results of neurodevelopmental examinations of two groups of premature infants at 6 months' corrected age. The difference in neurological development at 6 months' corrected age was statistically significant when comparison was corrected for premature infants who had neonatal seizures and periventricular hemorrhage. PMID- 14762588 TI - Brainstem auditory evoked response in normal term neonates. AB - Brainstem auditory evoked response (BAER) is a reliable test for neonatal auditory and neurological dysfunction and it permits early diagnosis and rehabilitation. The purpose of this study is to demonstrate latencies of BAER in normal term neonates in order to obtain reference values in a university hospital. BAER was performed in the second day of life in 47 normal newborns (25 male, 22 female) which gestational ages were higher than 37 and lower than or equal to 40 weeks that did not present familial history of deafness. The exam was performed with 80 dBHL alternating polarity 10/sec clicks presented monaurally. Two thousand stimulus trials were averaged and duplicated for each ear. Mean wave latencies in msec was: I, 1.79 (SD 0.20); II, 2.88 (SD 0.28); III, 4.54 (SD 0.31); IV, 5.86 (SD 0.36); V, 6.75 (SD 0.38); inter-peak latencies (IPL) I-III, 2.75 (SD 0.36); IPL III-V, 2.22 (SD 0.22); and IPL I-V, 4.97 (SD 0.43). PMID- 14762589 TI - Rett syndrome: clinical and epidemiological aspects in a Brazilian institution. AB - Rett syndrome (RS) is a neurodevelopmental disorder, preferentially found in females and specifically involving the functions on which intelligence and its expression depend--learning, hand use and speech--leaving many others intact. Mutations have been identified at Xq28 on the MECP2 gene (methyl-CpG 2), which selectively silences the expression of other genes whose location is still unknown. This is a study on clinical, diagnostic and epidemiological aspects of RS in a Brazilian sample. It included 33 female patients with chronic encephalopathy without known etiology. RS was diagnosed in 24 patients (72.7%): 17 (70.8%) had classical RS; 5 (20.8%), atypical RS and 2 (8.4%), potential RS. In 9 girls clinical data and/or laboratory studies excluded diagnosis of RS. Among the atypical RS patients, 4 were form fruste and one, congenital form. Among the girls with other encephalopathies, cerebral malformation was the most frequent finding. PMID- 14762590 TI - Brazilian standardised norms for a set of pictures are comparable with those obtained internationally. AB - Snodgrass & Vanderwart (1980) standardized a set of 260 pictures in the USA for use in studies of cognitive processes that employ pictured objects as laboratory analogues of object themselves. Since then similar norms for this set were obtained in Britain, Spain, Japan and Iceland and a larger set of 400 pictures (including the original 260: Cycowicz et al., 1997) was studied in France and Brazil. The present article provides a comparison of the norms obtained in Brazil and internationally. The pattern of correlations among the Brazilian and other standardizations were equivalent to that previously observed: despite pictures being judged to be of similar familiarity and visual complexity (high positive correlations), name agreement was less correlated, possibly due to differences in the languages spoken in each country and/or in the sample size used in each study. Results confirm the adequacy of the Brazilian norms. PMID- 14762591 TI - Memory complaints and test performance in healthy elderly persons. AB - In order to compare the use of a structured self-report questionnaire with direct questioning about memory problems, 71 healthy and independent aged individuals (63 women) from the community without risk factors for cognitive deficits were objectively asked about subjective memory complaints (SMC), given the Memory Complaint Questionnaire (MAC-Q) and then submitted to the Rey Auditory Verbal Learning Test (RAVLT). SMC positively correlated with higher scores on MAC-Q, although a significant percentage of the sample had SMC and lower scores on MAC-Q and also no SMC and higher scores on MAC-Q. Performance on RAVLT was significantly worse (p<0.05) for the group presenting SMC but not for the group with higher scores on the MAC-Q. We conclude that direct questioning maybe more clinically significant than a self report questionnaire, at least for elderly persons from the community without risk factors for cognitive decline or depression. PMID- 14762592 TI - Prevalence of potentially reversible dementias in a dementia outpatient clinic of a tertiary university-affiliated hospital in Brazil. AB - The importance of investigating the etiology for dementia lies in the possibility of treating potentially reversible dementias. The aims of this retrospective study are to determine the prevalence of potentially reversible dementias among 454 outpatients seen at the Cognitive and Behavioral Neurology Unit, Hospital das Clinicas, Sao Paulo University School of Medicine-Brazil, between the years of 1991 and 2001, and observe their evolution in follow-up. Among the initial 454 patients, 275 fulfilled the DSM-IV criteria for dementia. Alzheimer's disease was the most frequent diagnosis (164 cases; 59.6%). Twenty-two cases (8.0%) of potentially reversible dementia were observed, the most frequent diagnoses being neurosyphilis (nine cases) and hydrocephalus (six cases). Full recovery was observed in two patients and partial recovery in 10 patients. Two cases were not treated and eight cases were lost on follow-up. The prevalence found in the present study falls within the range reported in previous studies (0-30%). PMID- 14762593 TI - Frontotemporal dementia with severe thalamic involvement: a clinical and neuropathological study. AB - Frontotemporal dementia (FTD) is the third-leading cause of cortical dementia after Alzheimer's disease and Lewy body dementia, and is characterized by a dementia where behavioral disturbances are prominent and appear early in the course of the disease. We report the case of a 58 year-old man affected by dementia with behavioral disturbances, in addition to rigid-hypokinetic and a lower motor neuron syndrome that were present at later stages of the illness. Neuroimaging studies showed frontotemporal atrophy. Neuropathological studies revealed intense thalamic neuronal loss and astrocytic gliosis, as well as moderate frontotemporal neuronal loss, astrocytosis and spongiform degeneration. Thalamic degeneration has previously been described among the wide group of neuropathological features of FTD. The aim of the present study is to show the clinical and neuropathological aspects of thalamic degeneration in FTD, along with its role in behavioral disturbances, a common finding in this condition. PMID- 14762594 TI - Nightmares without atonia as an early symptom of diffuse Lewy bodies disease. AB - A male 70 years old patient with diffuse or "pure" Lewy body disease is described. The diagnosis was made based on clinical features of nightmares with no atonia, attention deficits with fluctuation in cognitive function, incapacity to find his way around the neighbourhood and other formerly familiar environments and mild neuropsychiatric symptoms. Neuropsychological assessment showed memory deficits, visuospatial and visuo-constructive disturbances. He had neither parkinsonism nor recurrent visual hallucinations typically well formed and detailed. Neuroimaging (computed tomography and magnetic resonance spectroscopy) showed mild diffuse cortical atrophy, mostly on the left temporal lobe and a decrease of N-acetyl-aspartate levels. A cholinesterase inhibitor was prescribed to this patient during 6 months with clinically relevant behavioral effect. Diagnosis confirmation was made by post-mortem neuropathological findings. Macroscopical features were mild atrophy on the frontal, parietal and temporal lobes, notedly on the frontal lobes. Microscopically, there was neuronal loss and diffuse classic Lewy bodies. Brainstem (substantia nigra, raphe nucleus, locus coeruleus, pedunculopontine nucleus), limbic cortex, and neocortex (frontal, parietal and temporal) were the areas of predilection for Lewy bodies. Hematoxylin-eosin and Bielschowsky staining did not show neuronal swelling (ballooned cell), argyrophilic inclusion (Pick's bodies), neurofibrillary tangles nor senile plaques. Immunohistochemical staining for anti-tau, anti-beta-amyloid, and anti-prion protein were negative. Antiubiquitine reaction was positive for Lewy body in the cerebral cortex and brainstem. PMID- 14762595 TI - Corpora amylacea in temporal lobe epilepsy associated with hippocampal sclerosis. AB - Hippocampal sclerosis (HS) is the commonest pathology in epileptic patients undergoing temporal lobe epilepsy surgery. Beside, there are an increased density of corpora amylacea (CA) founded in 6 to 63% of those cases. OBJECTIVE: verify the presence of CA and the clinical correlates of their occurrence in a consecutive series of patients undergoing temporal surgery with diagnosis of HS. METHOD: We reviewed 72 hippocampus specimens from January 1997 to July 2000. Student's t test for independent, samples, ANOVA and Tukey test were performed for statistical analysis. RESULTS: CA were found in 35 patients (49%), whose mean epilepsy duration (28.7 years) was significantly longer than that group of patients without CA (19.5 years, p=0.001). Besides, when CA were found, duration was also significantly correlated with distribution within hippocampus: 28.7 years with diffuse distribution of CA, 15.4 with exclusively subpial and 17.4 years with distribution subpial plus perivascular (p=0.001). CONCLUSION: Our findings corroborate the presence of CA in patients with HS and suggest that a longer duration of epilepsy correlate with a more distribution of CA in hippocampus. PMID- 14762596 TI - Myopathy of distal lower limbs: the clinical variant of Miyoshi. AB - Miyoshi distal dystrophy is a rare myopathy characterized by an autosomal recessive pattern of inheritance and it is prevalent in Japan. Onset of disease is in early adult life with weakness and atrophy of the leg muscles. Recently gene linkage to chromosome 2p12-14 has been established. We report three sisters, born of consanguineous parents. All of them noticed weakness and atrophy of leg muscles, and could not walk on their heels. In all of them the creatine kinase concentrations were very high. The electromyography showed myopathic patterns and the muscle biopsy disclosed dystrophic changes and an absence of dysferlin. There are few cases reported of Miyoshi distal dystrophy in Latin America. The Miyoshi myopathy may be distinct among the hereditary distal myopathies. PMID- 14762597 TI - [A retrospective study of Creutzfeldt-Jakob disease in North of Portugal 1993 2002: demographic, clinical and neuropathological features]. AB - OBJECTIVE: Description of the demographic, clinical and neuropathological features of 11 cases of Creutzfeldt-Jakob disease (CJD). METHOD: Review of the clinical and neuropathological features of patients with CJD diagnosed in hospitals in the North of Portugal between 1993 and 2002. RESULTS: Eleven patients were identified, 4 females: mean age of onset of symptoms--64 years, mean duration of disease--8 months. All presented with a syndrome of progressive dementia with myoclonus, with four patients presenting with cerebellar signs. Neuropathological examination of brain at autopsy showed spongiosis and reactive gliosis associated with neuronal loss. In eight cases immunocytochemistry for prion protein (PrP) was carried out and was positive. CONCLUSION: The group of patients described represents the heterogeneity of the clinical phenotypes possible in CJD. Neuropathological examination is still indispensable to make the definitive diagnosis of the disease. PMID- 14762598 TI - [Anatomical and functional abnormalities of central nervous system in autistic disorder: a MRI and SPECT study]. AB - We present a study of anatomical and functional abnormalities of central nervous system (CNS) from patients with autistic disorder (AD); magnetic resonance imaging (MRI) and single photon emission computed tomography (SPECT) were used for the investigation. The population studied was composed of 24 patients, 15 (62.5%) males and 9 (17.5%) females, mean age 9 years. MRI was performed in all patients and SPECT was performed in 19 patients; 75% (n=18) of patients had anatomical abnormalities and all patients that realized SPECT had functional abnormalities. Anatomical abnormalities were preferentially noted in corpus callosum (25%), septum pellucidum (15.63%), cerebral ventricles (12.55%), cerebellum (9.38%), temporal lobes (6.25%), occipital lobes (6.25%) and hippocampus (6.25%). Functional abnormalities predominated in frontal lobes (53.13%), temporal lobes (28.13%), parietal lobes (15.63%) and basal ganglia (3.13%). However, anatomical and functional abnormalities of CNS are not priorities for diagnosis, which should have always clinical validation. PMID- 14762599 TI - Effect of acetyl-L-carnitine on VIP-ergic neurons in the jejunum submucous plexus of diabetic rats. AB - The effect of the treatment with acetyl-L-carnitine (ALC) on neurons releasing the vasoactive intestinal polypeptide (VIP) of the submucous plexus in the jejunum of diabetic rats was the purpose of our investigation. Diabetes (DM) was induced by injecting streptozotocin endoveneously (35 mg/kg). After sacrificing the animals, the jejunum was collected and processed for VIP detection. Four groups were used: C (non-diabetic), CC (non-diabetic treated with ALC), D (diabetic), DC (diabetes treated with ALC). We analyzed the immunoreactivity and the cellular profile of 126 cell bodies. The treatment with ALC improved some aspects of DM. However, it promoted a small increase in the area of neurons from group CC, suggesting a possible neurotrophic effect. Neurons from groups D and DC showed a large increase in their cellular profile and immunoreactivity when compared to C and CC, suggesting a larger concentration of this neurotransmitter within the neurons that produce it. This observation constitutes a recurrent finding in diabetic animals, suggesting that ALC does not interfere in the pathophysiological mechanisms that unchain a higher production and/or neurotransmitter accumulation and increase the profile of the VIP-ergic neurons. PMID- 14762600 TI - [Determination of autoantibody for myelin antigens in the serum of patients HLA DQB1*0602 with multiple sclerosis]. AB - Multiple sclerosis (MS) is a chronic inflammatory demyelinating disease of the human central nervous system (CNS) mediated by autoimmune Th1 lymphocytes. We determined the serum levels of autoantibodies for myelin basic protein (MBP), proteolipid (PLP) and myelin oligodendrocyte glycoprotein sequence MOG 92-106 in a group of 54 healthy individuals and 26 MS patients expressing or not HLA DQB1*0602. Regardless expression of the susceptibility allele DQB1*0602, MS patients presented marked (p<0.0001) IgG antibody production for MBP and MOG92 106. Yet, significant (p<0.0001) IgA antibody levels were mainly observed for PLP and MOG antigens. Our results suggest that other HLA class II alleles may be conferring susceptibility to MS in this population and influencing the pattern of immune recognition of encephalitogen antigens. Furthermore, distinct IgG and/or IgA autoantibody production may be contributing to the control or maintenance of the CNS inflammatory reaction. PMID- 14762601 TI - [Benign childhood epilepsy with centrotemporal spikes: high and low central focus]. AB - Twenty children with benign childhood epilepsy with centrotemporal spikes were studied. Anamnesis, neurological exam, "Columbia" Scale (1993) application and digital electroencephalogram were carried out. The digital electroencephalogram was recorded with electrodes according to 10-20 international system, and a supplementary electrode, between C3 and T3 or C4 and T4-C5 or C6, respectively, at the side with a higher number of spikes. The averaging of the spikes was carried. Localization and the laterality of the spikes were analyzed. Seizures involving a superior limb occurred in a higher proportion of cases with spikes of maximum negativity in C3, C4. Independent epileptiform activity, different from centrotemporal, was more frequent in the cases were the maximum negativity of the spikes occurred in C3 or C4 regions. There are clinical and electroencephalographic differences according to the localization of the discharges PMID- 14762602 TI - [Sleep disorders in temporal lobe epilepsy]. AB - The objective of this study was to evaluate sleep macrostructure and sleep disturbance in a group of 39 patients with temporal lobe epilepsy (TLE). Patients completed questionnaires to evaluate their sleep and subjective daytime sleepiness (Epworth Sleepiness Scale [ESS]) and undergone Polysomnography and Multiple Sleep Latency Test (MSLT). Daytime sleepiness was the most frequent complaint (85%), followed by wakefulness during sleep, history of seizures during sleep (75%) and initial insomnia (26%). Parasomnias (67%), obstructive sleep apneas (13%), restless legs syndrome (15%) and periodic limb movements (5%) were the most frequent sleep disorders. The most frequent changes of sleep patterns were: sleep architecture fragmentation (100%), decreased amount of REM sleep (92%) and increase in time awake after sleep onset (77%). There were significative correlations between the ESS and the MSLT (p<0,05). In conclusion, TLE patients have fragmented sleep with increased sleep stages shifts, increased number of awakenings and in time awake after sleep onset. REM sleep was decreased. Daytime sleepiness was the most frequent complaint in TLE patients. PMID- 14762603 TI - [Vigabatrin in the treatment of epilepsy in patients with West syndrome and tuberous sclerosis]. AB - PURPOSE: To report the efficacy of vigabatrin in seizures control, as well as the electroencephalographic abnormalities in children with tuberous sclerosis and West syndrome. METHOD: Retrospective study, with clinical, neuroimaging, and electroencephalographic data. RESULTS: Seven patients were followed, and the median time of follow-up was 10 years. Four of them were females and all were white. The mean age of seizures onset was 3.4 months. All patients used antiepileptic drugs associations, at least 2 drugs each therapeutic scheme, each one of the patients have used at least two different schemes. Vigabatrin as monotherapy or adjuvant was started in a mean age of seven years or 4 years after the onset of symptoms. Five from seven patients on vigabatrin became seizure free. CONCLUSION: Vigabatrin was efficient in seizures control, leading to a better prognosis. PMID- 14762604 TI - [Depressive symptoms and anxiety in patients with chronic and episodic tension type headache]. AB - BACKGROUND: Tension-type headache (TTH) is recognized as the most prevalent type of headache. Despite this, there is a limited understanding of the entity's physiology, epidemiology, and clinical presentation. Anxiety and depression are recognized comorbidities present among patients with TTH. OBJECTIVE: To quantify the prevalence of anxiety and depression among patients with episodic and chronic TTH. METHOD: fifty patients with episodic TTH and fifty patients with chronic TTH completed Beck's anxiety and depression surveys. Only patients presenting with moderate to severe scores were considered. RESULTS: among the patients with episodic TTH, anxiety and depression were observed in 30 (60%) and 16 (32%) patients respectively. Among the patients with chronic TTH, anxiety was observed in 22 (44%) patients, and depression was observed in 20 (40%). CONCLUSION: both comorbidities are important among patients with episodic and chronic TTH. Neglecting this association may result in failure of symptomatic and prophylactic treatment ultimately leading to lost quality of life. PMID- 14762605 TI - [Acoustic analysis of prosody in females with Parkinson's disease: effect of L dopa]. AB - BACKGROUND: Untreated Parkinson's disease (PD) patients display speech abnormalities characterized by narrow range of fundamental frequency (Fo) variation and higher amplitude. The role of dopaminergic systems in the pathogenesis of these changes as well as the therapeutic effect of L-dopa have not been clearly determined. The aim of this study is to assess the effect of L dopa on the prosodic features of the speech of female PD patients. METHOD: We have studied 8 PD female patients (63.5 +/- 6.8 years). The PD patients (H-Y stage 2, seven subjects; stage 2.5, four patients; stage 1, one patient) were examined when off and on. The fundamental frequency, intensity and duration of five affirmative sentences were analysed with the software WinPitch 1.8 (Philippe Martin). The analysed Fo parameters were: usual Fo (Fo us), maximal Fo (Fo max), minimal Fo (Fo min), Fo max of the prenuclear tonic (Fo max PNT), Fo min of the prenuclear tonic (Fo min PNT), Fo max of the nuclear tonic (Fo max NT), Fo min of the nuclear tonic (Fo min TN), velocity of melodic variation of the PNT (VPNT), velocity of melodic variation of the NT (VNT), amplitude of melodic variation of the PNT (APNT), and amplitude of melodic variation of the NT (ANT). Duration of the statement (D), duration of the PNT (DPNT), duration of the NT (DNT) and number of syllables per second (NSS) were the duration parameters investigated. The following intensity variables were studied: mean intensity (MI), maximal intensity (I max), minimal intensity (I min), intensity of the PNT (IPNT) and intensity of the NT (INT). Differences were statistically significant if p<0.05. RESULTS: When on, PD patients had significantly higher Fo max PNT (201.9 +/- 27.5 Hz), higher APNT (22.1 +/- 18.7 Hz), and smaller MI (32 +/- 6.2 dB) in comparison with off period (respectively: 168.5 +/- 83.5 Hz, 20.4 +/- 12.1 Hz, 32.7 +/- 5.6 dB). CONCLUSION: Our findings show that L-dopa replacement therapy slightly improves the Fo range and the intensity of speech of Brazilian female PD patients. These results suggest that non-dopaminergic systems are probably involved in the pathogenesis of prosody changes in PD. They also indicate that therapies other than dopaminergic agents are warranted to treat speech abnormalities in PD. PMID- 14762606 TI - [Acoustic analysis of prosody in females with Parkinson's disease: comparison with normal controls]. AB - BACKGROUND: Voice abnormalities, collectively labeled as hypokinetic dysarthria, have been well characterized and occur in 90% of Parkinson's disease (PD) patients. However, studies of speech in PD, particularly of patients whose native language is other than English, are rarely found. The aim of this study is to compare the prosodic features of the speech of female PD patients and gender- and age-matched controls using acoustic analysis. METHOD: We have studied 8 PD female patients (68.4 +/- 6.4 years) and 8 female controls (63.5 +/- 6.8 years). The PD patients (H-Y stage 2, one subject; stage 2.5, four patients; stage 3, three patients), although treated with L-dopa, were examined when off. The fundamental frequency (Fo), intensity and duration of the speech were analysed with the software WinPitch 1.8 (Philippe Martin). The analysed Fo parameters were: usual Fo (Fo us), maximal Fo (Fo max), minimal Fo (Fo min), Fo max of the prenuclear tonic (Fo max PNT), Fo min of the prenuclear tonic (Fo min PNT), Fo max of the nuclear tonic (Fo max NT), Fo min of the nuclear tonic (Fo min TN), velocity of melodic variation of the PNT (VPNT), velocity of melodic variation of the NT (VNT), amplitude of melodic variation of the PNT (APNT), and amplitude of melodic variation of the NT (ANT). Duration of the statement (D), duration of the PNT (DPNT), duration of the NT (DNT) and number of syllables per second (NSS) were the duration parameters investigated. The following intensity variables were studied: mean intensity (MI), maximal intensity (I max), minimal intensity (I min), intensity of the PNT (IPNT) and intensity of the NT (INT). Differences were statistically significant if p<0.05. RESULTS: PD patients had significantly lower Fo max (232.1 +/- 32.9 Hz), lower Fo max PNT (168.5 +/- 83.5 Hz), lower Fo max NT (182.1 +/- 29.5 Hz), lower VNT (0.2 +/- 0.1 Hz), smaller APNT (20.4 +/- 12.1 Hz), smaller ANT (26 +/- 12.6 Hz), smaller NSS (4.8 +/- 0.1), higher IM (32.7 +/- 5.6 dB), higher I max (37.1 +/- 2.9 dB), higher I min (23.2 +/- 6.1 dB) and higher IPNT (35.4 +/- 2.9 dB) than controls (respectively: 248.7 +/- 23.9 Hz, 216.4 +/- 25.6 Hz, 197.5 +/- 35.8 Hz, 0.3 +/- 0.2 Hz, 22.7 +/- 15.9 Hz, 36.3 +/- 21.8 Hz, 5.3 +/- 0.8, 30.4 +/- 6.1 dB, 20.2 +/- 6.6 dB, 33.1 +/- 2.9 dB, and 30.9 +/- 4.3 dB. CONCLUSION: In comparison with controls, Brazilian female PD patients have a slower speech characterized by a narrow range of Fo variation (which likely accounts for the poor vocal tessitura of parkinsonian speech) and higher intensity. The latter is consistent with a compensatory attempt to overcome the latter. PMID- 14762607 TI - [Clinical and therapeutic aspects of prolactinoma in men]. AB - Macroprolactinomas predominate in males in comparison to microprolactinomas, with greater trend to invasiveness than in females. The clinical treatment has been the first option to prolactinomas, in both macro and microadenomas, irrespective the sex. We compared clinical presentation, prolactin levels, neuroradiologic invasiveness and prolactinemia response of 23 men with prolactinomas subjected to clinical therapy (group 1) with 19 who went also through surgical and/or radiotherapeutic treatment (group 2). The statistical analysis was done by the tests of chi-square or exact of Fisher, in order to compare proportions, and by t of Student or Mann-Whitney, in order to compare means. The level of significance adopted was 5% (p<0.05). The two groups were similar regarding age (p=0.23), period between start of the first symptom and diagnosis (p=0.82), prolactin levels before treatment (p=0.41) and invasive macroadenomas proportion (p=0.096). There was significantly greater percentage of headache (p=0.009) and visual deficit (p=0.025) in group 2, as well as the drug usage (p=0.007) and observation (p=0.0005) periods were superior in this group. The variations of prolactin levels before and after therapy (p=0.49) as well as the percentage of prolactin normalization (p=0.20) did not show any significant difference when comparing the two groups. We conclude, emphasizing the relevance of precocious prolactinoma diagnostic in men, because of the demonstrated morbidity. We strengthen the use of dopamine agonist as the first therapeutic option irrespective the adenoma size. PMID- 14762608 TI - [Chronic subdural hematoma: study of 161 patients and the relationship with coagulation abnormalities]. AB - The chronic subdural hematoma represents one of the most frequent types of intracranial hemorrhage. One hundred sixty one patients operated in the Clinical Hospital of State University of Campinas-SP (UNICAMP) from April 1994 to May 2000, were analyzed retrospectively in order to characterize the population and to study the importance of the pre-operative coagulation analysis in the outcome. The majority of the population was male (86%), white (85.1%) and in the fifth decade (median age: 57 years). A high mortality index in the postoperative period was found in patients with INR (international normalized ratio) values greater than 1.25 and/or thrombocytopenia (p<0.001 and p=0.004 respectively). Patients with previous head injury history (76%) showed a lower mortality (p=0.044). The results stand out the importance of the pre-operative evaluation with coagulation studies in order to correct possible abnormalities. PMID- 14762609 TI - [Ischaemic stroke in two children with HIV-1]. AB - Cerebral ischaemia caused by inflammatory vasculopathies has been described as a complication of human immunodeficiency virus (HIV) infection. The goal of our study is to report two cases of pediatric human immunodeficiency virus infection and cerebrovascular manifestations. We describe two pre-school boys, from a group of 204 outpatients, who presented fever, seizures, hemiparesis and impairment of conscience level as a first symptom of HIV-1 infection. The serial imaging studies revealed infarction of middle cerebral artery in both cases. The first one child had a severe spastic tetraparesis and partial epilepsy and died four years later without any improvement despite of the antiretroviral therapy. The second patient had a right hemiparesis and global aphasia totally recovered two years later with antiretroviral and rehabilitation therapies. HIV infection should be included in differential diagnosis of children who present with seizures, mental status change or focal neurological deficits and seizures. PMID- 14762610 TI - Sudden sensorineural hearing loss: a case report supporting the immunologic theory. AB - Sudden sensorineural hearing loss (SSNHL) is one of the autoimmune diseases of the inner ear (AIED), which is characterized by a hearing loss of above 30 decibels in at least three contiguous audiometric frequencies over a time course of 72 hours or shorter. Its cause can be found in only 10% to 15% of patients. Histopathologic findings have reported retrograde neuronal degeneration and atrophy of Corti's organ and of the vascular stria. This paper describes a case of a middle-aged female patient undergoing a treatment for hyperthyroidism who developed bilateral SSNHL. The patient was treated with methylprednisone (1 mg/kg/day) for three days with considerable hearing improvement. This treatment was followed by lung and kidney tuberculosis. The immune mechanism of this entity and the possibility of interconnected participation of the antigen type, of an autoimmune disease and of bacterial infection are discussed. PMID- 14762611 TI - Hemophilia and child abuse as possible causes of epidural hematoma: case report. AB - INTRODUCTION: Head trauma is an important consequence of child abuse. Specific pathophysiological mechanisms in child abuse are responsible for the "whiplash shaken-baby syndrome", which would favour the occurrence of intracranial hemorrhages. CASE REPORT: We report the case of a child who developed epidural hematoma following minor-intensity head trauma. Initial diagnosis of child abuse was made, but subsequent investigation led to the diagnosis of hemophilia A. CONCLUSION: Even though epidural hematoma is not closely associated with child abuse, this etiology must always be considered when the reported trauma mechanism is out of proportion to the magnitude of the encountered lesions. PMID- 14762612 TI - Cerebral sinovenous thrombosis in a nephrotic child. AB - Nephrotic syndrome in infancy and childhood is known to be associated with a hypercoagulable state and thromboembolic complications, but cerebral sinovenous thrombosis (CST) is a very rare and serious one, with only a few isolated reports in the literature. A case is presented of a 9-year-old boy with nephrotic syndrome that acutely developed signs and symptoms of intracranial hypertension syndrome. CST was diagnosed on cranial CT and MRI and he gradually recovered after treatment with anticoagulants. The diagnosis of CST should be considered in any patient with nephrotic syndrome who develops neurologic symptoms. The discussion of this case, coupled with a review of the literature, emphasizes that early diagnosis is essential for institution of anticoagulation therapy and a successful outcome. This report also illustrates the difficulties that may be encountered in managing such a patient. PMID- 14762613 TI - Peritoneal dissemination from central neurocytoma: case report. AB - OBJECTIVE: Central neurocytoma is a low grade tumor of neuroglial origin and a relatively new histological entity. Only a few cases have been reported and its biological behavior is still uncertain. Some cases have shown an aggressive behavior (local recurrence, malignant dedifferentiation or CSF dissemination) and challenged the initial view of its relative benignity. A case of central neurocytoma with peritoneal dissemination is presented. CASE: A six years-old boy with recurrent neurocytoma of III ventricle and left thalamus showed fast growth of tumor rest and ascites three and a half years after subtotal removal of the lesion. Tumor cells were identified in the ascitic fluid and implanted in the peritoneum. Chemotherapy was initiated immediately after diagnosis of peritoneal dissemination (etoposide, carboplatin, doxorubicin and cyclophosphamide). The patient developed metabolic imbalance and respiratory failure due to rapid formation of ascitic fluid and died 3 days after the diagnosis of peritoneal dissemination was established. CONCLUSION: Central neurocytoma is a low grade tumor with low values of the proliferative index in the majority of cases. In spite of that, some tumors may present a very aggressive behavior and extraneural dissemination. Evaluation of proliferative index may be a guideline parameter for planning adjuvant therapies after surgical treatment in selected cases. Extraneural dissemination may occur in some cases specially in patients with ventriculoperitoneal shunt. PMID- 14762614 TI - Cellular schwannoma: a rare spinal benign nerve-sheath tumor with a pseudosarcomatous appearance: case report. AB - We report a case of cellular schwannoma, a rare benign nerve-sheath tumor in a 27 year-old woman. It was presented as a voluminous lesion in the paraspinal region that caused lumbar vertebral body destruction. These features, in association to the microscopic aspects of a hypercellular, pleomorphic neoplasm may lead to a false impression of a malignant tumor. Therefore, it is important to have an accurate examination to confirm the benign nature of this tumor thus avoiding unnecessary therapy. PMID- 14762615 TI - Traumatic bilateral intraorbital (subperiosteal) hematoma associated with epidural hematoma: case report. AB - Extradural hematoma (EDH) is a frequent lesion, with an incidence varying from 0.2 to 6% in patients admitted to hospital due to traumatic head injury. The higher incidence is found in patients with more severe injuries. The association of EDH with subperiosteal intraorbital hematomas is rarely reported, and we were not able to find in the literature any report of traumatic bilateral intraorbital hematomas and EDH. We report this case of a 32 year-old man with bilateral intraorbital (subperiosteal) hematoma associated with unilateral EDH. The lesions were treated surgically, but unfortunately with an unfavorable outcome. PMID- 14762616 TI - [Cerebral and systemic venous thrombosis associated to prothrombin G20210 mutation: case report]. AB - Cerebral venous thrombosis is a clinical condition of difficult diagnosis, and poor prognosis when treatment is not started early. There is a long list of causes, and hereby we describe a case associated to prothrombin G20210 mutation. A 53-year-old man, white, was admitted with status epilepticus. After seizures control, he developed intracranial hypertension, with headache and vomiting, and bilateral papilledema. His past medical and familial history were unremarkable. He was a nonsmoker, no drug and alcohol user. CT scan and MRI showed right temporal and parietal infarct with hemorrhagic transformation. Spinal tap with opening pressure of 500 mmH2O showed normal CSF examination. MRI angiography disclosed superior sinus, right transverse and sigmoid sinus complete thrombosis. He was started with heparin and oral warfarin. In spite of anticoagulation, two months later he developed deep right inferior limb thrombosis. All the initial tests were normal, and test for prothrombin G20210 mutation was positive. He needed a much higher than conventional daily dose of warfarin to keep him asymptomatic. PMID- 14762617 TI - [Treatment of epilepsy: consensus of the Brazilian specialists]. AB - Epilepsy is a frequent condition in the world. Recently a study in Brazil showed prevalence of 18/1000 inhabitants in Sao Jose do Rio Preto, Sao Paulo State. In the last decade, new therapeutic options were discovered or developed. The main therapeutic decision method is based on randomized clinical trials. This method represents the higher level of evidence. However, even these studies have limitations and in some cases the treatment of choice remains controversial. In these instances, the epilepsy experts' opinions become helpful. In 2001 a similar study had been conducted in USA. The aim of this study is to create guidelines for epilepsy treatment based on the opinion of the Brazilian experts. These guidelines can be used to create manuals and strategies for the treatment of some epileptic syndromes according to Brazilian experts. As compared to the North American guidelines our study better reflects the resources available in our country. PMID- 14762618 TI - [Landmarks to the cranial approaches]. PMID- 14762619 TI - [Complication of ventriculoperitoneal shunting: inguinal hernia with scrotal migration of catheter]. PMID- 14762626 TI - Cryptococcosis: a review of the Brazilian experience for the disease. AB - Cryptococcosis is a systemic mycosis caused by Cryptococcus neoformans. The disease occurs in patients with cellular immunodeficiency. The incidence of cryptococcosis arises with aids, and mycosis is one of the opportunistic infections that defines AIDS. After the HAART era the occurrence of cryptococcosis decreased all over the world, but it still continues to be a prevalent disease in Brazil. Thus, we consider this paper to be very important as a result of our reviewing of Brazilian literature regarding some relevant aspects of that disease. PMID- 14762628 TI - Pancreatic involvement in fatal human leptospirosis: clinical and histopathological features. AB - Hyperamylasemia has been reported in more than 65% of patients with severe leptospirosis, and the true diagnosis of acute pancreatitis is complicated by the fact that renal failure can increase serum amylase levels. Based on these data we retrospectively analyzed the clinical and histopathological features of pancreas involvement in 13 cases of fatal human leptospirosis. The most common signs and symptoms presented at admission were fever, chills, vomiting, myalgia, dehydratation, abdominal pain and diarrhea. Trombocytopenia was evident in 11 patients. Mild increased of AST and ALT levels was seen in 9 patients. Hyperamylasemia was recorded in every patient in whom it was measured, with values above 180 IU/L (3 cases). All patients presented acute renal failure and five have been submitted to dialytic treatment. The main cause of death was acute respiratory failure due to pulmonary hemorrhage. Pancreas fragments were collected for histological study and fat necrosis was the criterion used to classify acute pancreatitis. Histological pancreatic findings were edema, mild inflammatory infiltrate of lymphocytes, hemorrhage, congestion, fat necrosis and calcification. All the patients infected with severe form of leptospirosis who develop abdominal pain should raise the suspect of pancreatic involvement. PMID- 14762630 TI - Immunoprecipitation techniques and Elisa in the detection of anti-Fonsecaea pedrosoi antibodies in chromoblastomycosis. AB - Chromoblastomycosis (CBM) is a chronic subcutaneous infection caused by several dematiaceous fungi. The most commonly etiological agent found in Brazil is Fonsecaea pedrosoi, which appears as thick walled, brownish colored cells with transverse and longitudinal division in the lesions, called "muriform cells". This disease is found worldwide but countries like Madagascar and Brazil have highest incidence. Diagnosis is made by clinical, direct and histopathologic examination and culture of specimens. Serological tests have been used to identify specific antibodies against Fonsecaea pedrosoi antigens, as well as immunotechniques have been used for CBM serological identification and diagnosis. In the present study double immunodiffusion (DID), counterimmunoelectrophoresis (CIE) and immunoenzymatic test (ELISA) have been used to evaluate humoral immune response in patients with CBM caused by F. pedrosoi. Metabolic antigen was used for immunoprecipitation tests (DID and CIE) while somatic antigen for ELISA. Our results demonstrated 53% sensitivity and 96% specificity for DID, while CIE presented 68% sensitivity and 90.5% specificity. ELISA demonstrated 78% sensibility and 83% specificity. Serological tests can be a useful tool to study different aspects of CBM, such as helping differential diagnosis, when culture of the pathogenic agent is impossible. PMID- 14762631 TI - Assessment of the rapid test based on an immunochromatography technique for detecting anti-Treponema pallidum antibodies. AB - A rapid test based on an immunochromatography assay - Determine Syphilis TP (Abbott Lab.) for detecting specific antibodies to Treponema pallidum was evaluated against serum samples from patients with clinical, epidemiological and serological diagnosis of syphilis, patients with sexually transmitted disease other than syphilis, and individuals with negative serology for syphilis. The Determine test presented the sensitivity of 93.6%, specificity of 92.5%, and positive predictive value and negative predictive value of 95.2% and 93.7%, respectively. One serum sample from patient with recent latent syphilis showed a prozone reaction. Determine is a rapid assay, highly specific and easy to perform. This technique obviates the need of equipment and its diagnostic features demonstrate that it may be applicable as an alternative assay for syphilis screening under some emergency conditions or for patients living in remote localities. PMID- 14762632 TI - Evaluation of an in-house specific immunoglobulin G (IgG) avidity ELISA for distinguishing recent primary from long-term human cytomegalovirus (HCMV) infection. AB - This article describes the standardization and evaluation of an in-house specific IgG avidity ELISA for distinguishing recent primary from long-term human cytomegalovirus (HCMV) infection. The test was standardized with the commercial kit ETI-CYTOK G Plus (Sorin Biomedica, Italy) using 8 M urea in phosphate buffered saline to dissociate low-avidity antibodies after the antigen-antibody interaction. The performance of the in-house assay was compared to that of the commercial automated VIDAS CMV IgG avidity test (bioM rieux, France). Forty-nine sera, 24 from patients with a recent primary HCMV infection and 25 from patients with a long-term HCMV infection and a sustained persistence of specific IgM antibodies, were tested. Similar results were obtained with the two avidity methods. All 24 sera from patients with recently acquired infection had avidity indices compatible with acute HCMV infection by the VIDAS method, whereas with the in-house method, one serum sample had an equivocal result. In the 25 sera from patients with long-term infection, identical results were obtained with the two methods, with only one serum sample having an incompatible value. These findings suggest that our in-house avidity test could be a potentially useful tool for the immunodiagnosis of HCMV infection. PMID- 14762633 TI - Assessment of chloroquine single dose treatment of malaria due to Plasmodium vivax in Brazilian Amazon. AB - Malaria regions of the Amazon basin have been characterized by difficult access and non-compliance of the patients to treatment. In an attempt to assess the schizonticide efficacy of chloroquine in a single dose of 600 mg, the authors realized a double-blind, placebo-controlled trial in 132 outpatients with vivax malaria. Patients were distributed into two groups: group CPLA, given chloroquine 600 mg (single dose) on the first day of treatment, and two doses of placebo on second and third days. Group CHLO, given chloroquine 600 mg on first day and 450 mg on second and third day. Geometric means of the parasite density during the follow-up was similar in both groups. No differences were observed in the parasitological cure between the two groups (p = 0.442). There was clinical and parasitological efficacy in treatment of patients given a single-dose of chloroquine. This suggests that its restricted use could be indicated in remote areas of Brazilian Amazon Region, nevertheless the inadequate response of three patients indicates the need for further studies. PMID- 14762635 TI - Cerebral mass lesion due to cytomegalovirus in a patient with AIDS: case report and literature review. AB - Cytomegalovirus (CMV) disease in acquired immunodeficiency syndrome (AIDS) patients most commonly presents as chorioretinitis and gastro-intestinal infection. Neurological involvement due to CMV may cause several clinical presentations: polyradiculitis, myelitis, encephalitis, ventriculo-encephalitis, and mononeuritis multiplex. Rarely, cerebral mass lesion is described. We report a 39 year-old woman with AIDS and previous cerebral toxoplasmosis. She presented with fever, seizures, and vulval ulcers. Her chest X-ray showed multiple lung nodules, and a large frontal lobe lesion was seen in a brain computed tomography scan. She underwent a brain biopsy through a frontal craniotomy, but her condition deteriorated and she died in the first postoperative day. Histopathological studies and immunohistochemistry disclosed CMV disease, and there was no evidence of cerebral toxoplasmosis, bacterial, mycobacterial or fungal infection. CMV disease should be considered in the differential diagnosis of cerebral mass lesion in AIDS patients. High suspicion index, timely diagnostic procedures (surgical or minimally invasive), and proper utilization of prophylactic and therapeutic medication could improve outcome of these patients. PMID- 14762636 TI - Comparison between immunomagnetic separation, coupled with immunofluorescence, and the techniques of Faust et al. and of Lutz for the diagnosis of Giardia lamblia cysts in human feces. AB - In the present study, the performance of Immunomagnetic Separation technique, coupled with Immunofluorescence (IMS-IFA), was compared with the FAUST et al. and Lutz parasitological techniques for the detection of Giardia lamblia cysts in human feces. One hundred and twenty-seven samples were evaluated by the three techniques at the same time showing a rate of cyst detection of 27.5% by IMS-IFA and 15.7% by both Faust et al. and Lutz techniques. Data analysis showed a higher sensitivity of IMS-IFA for the detection of G. lamblia cysts in comparison with the techniques of FAUST et al. and Lutz. The use of this methodology as a routine procedure enables the processing of many samples simultaneously, in order to increase recovery rate of G. lamblia cysts and reduce the time of sample storage. PMID- 14762637 TI - Stiffness and postural stability in adults with Down syndrome. AB - The purpose of this study was to characterize postural sway in quiet standing under eyes-open and eyes-closed conditions, and to obtain a measure of postural stiffness during quiet standing in adults with Down syndrome (DS) versus control subjects. We obtained descriptive measures from centre-of-pressure (COP) data and analysed and compared COP trajectories and postural stiffness estimates from two stochastic models, the "pinned polymer" (PP) and "inverted pendulum" (IP) models. These estimates were correlated with clinical measures of muscle tone. Our results showed that overall, estimated values for postural stiffness from both models were larger for the DS group than for normal controls. In addition, average stiffness measures were greater under the eyes-closed condition than under the eyes-open condition for the DS group. The IP model detected significant trends over trials whereas the PP model did not. Clinical assessment of muscle tone for the DS group ranged from low to high-normal and there was no significant correlation with the postural stiffness measures obtained from either model. These results suggest that individuals with DS have the ability to modulate their underlying "stiffness" under conditions of quiet standing. Furthermore, there appears to be no strong relationship between clinical measures of muscle tone and postural stiffness measures under dynamic conditions. PMID- 14762639 TI - Feedforward activity of the cervical flexor muscles during voluntary arm movements is delayed in chronic neck pain. AB - The objective of this study was to compare onset of deep and superficial cervical flexor muscle activity during rapid, unilateral arm movements between ten patients with chronic neck pain and 12 control subjects. Deep cervical flexor (DCF) electromyographic activity (EMG) was recorded with custom electrodes inserted via the nose and fixed by suction to the posterior mucosa of the oropharynx. Surface electrodes were placed over the sternocleidomastoid (SCM) and anterior scalene (AS) muscles. While standing, subjects flexed and extended the right arm in response to a visual stimulus. For the control group, activation of DCF, SCM and AS muscles occurred less than 50 ms after the onset of deltoid activity, which is consistent with feedforward control of the neck during arm flexion and extension. When subjects with a history of neck pain flexed the arm, the onsets of DCF and contralateral SCM and AS muscles were significantly delayed ( p<0.05). It is concluded that the delay in neck muscle activity associated with movement of the arm in patients with neck pain indicates a significant deficit in the automatic feedforward control of the cervical spine. As the deep cervical muscles are fundamentally important for support of the cervical lordosis and the cervical joints, change in the feedforward response may leave the cervical spine vulnerable to reactive forces from arm movement. PMID- 14762638 TI - A detailed analysis of the planning and execution of prehension movements by three adolescents with spastic hemiparesis due to cerebral palsy. AB - Assuming that primary symptoms of motor disorders can best be distinguished from signs of adaptation through behavioral analyses on an individual basis, the present study provides a detailed analysis of the prehension movements of three adolescents with mild spastic hemiparesis of different etiology. We investigated the extent to which the hemiparetic participants took their movement limitations into account when planning and performing sequences of prehension movements. We examined three indices of flexibility in grip planning in conjunction with an analysis of arm-joint coordination patterns as the movements unfolded. Participants were asked to repeatedly grasp a square object of which the position was gradually changed leftwards or rightwards. In half the trials the goal of the task was to lift the object, in the other half it had to be rotated back-and forth. Trunk, arm, and hand movements were recorded with two synchronized 3-D motion-tracking systems. The movements of the hemiparetic participants were compared with the average performance of 11 control participants of which the collective data were taken to represent a typical control participant. Whereas one hemiparetic participant (GV) maintained a single grasping pattern throughout the experiment, the other two (CV and LC) only partially persevered in previously adopted grasping patterns. The shoulder contributed more and the wrist contributed less to this perseverance. No effects of task goal were found on grip selection. However, two hemiparetic participants (CV and LC) did tune their hand displacement to the task that followed the grasps. Taken collectively, the results show that the hemiparetic participants took their limitations into account when performing movements, but not when planning movements. PMID- 14762640 TI - Airborne chemistry: acoustic levitation in chemical analysis. AB - This review with 60 references describes a unique path to miniaturisation, that is, the use of acoustic levitation in analytical and bioanalytical chemistry applications. Levitation of small volumes of sample by means of a levitation technique can be used as a way to avoid solid walls around the sample, thus circumventing the main problem of miniaturisation, the unfavourable surface-to volume ratio. Different techniques for sample levitation have been developed and improved. Of the levitation techniques described, acoustic or ultrasonic levitation fulfils all requirements for analytical chemistry applications. This technique has previously been used to study properties of molten materials and the equilibrium shape()and stability of liquid drops. Temperature and mass transfer in levitated drops have also been described, as have crystallisation and microgravity applications. The airborne analytical system described here is equipped with different and exchangeable remote detection systems. The levitated drops are normally in the 100 nL-2 microL volume range and additions to the levitated drop can be made in the pL-volume range. The use of levitated drops in analytical and bioanalytical chemistry offers several benefits. Several remote detection systems are compatible with acoustic levitation, including fluorescence imaging detection, right angle light scattering, Raman spectroscopy, and X-ray diffraction. Applications include liquid/liquid extractions, solvent exchange, analyte enrichment, single-cell analysis, cell-cell communication studies, precipitation screening of proteins to establish nucleation conditions, and crystallisation of proteins and pharmaceuticals. PMID- 14762643 TI - Prompt-gamma neutron activation analysis facility for in vivo body composition studies in small animals. AB - The design, calibration, dosimetry and performance evaluation of a prompt-gamma neutron activation analysis facility for in vivo body composition studies in small animals (i.e. rats or rabbits) is discussed. The system design was guided by Monte Carlo transport calculations using MCNP-4C code. A system was built and performance evaluation was made using a 185-GBq Pu-Be neutron source. Prompt gamma rays produced by neutron capture reactions were detected by a combination of a NaI(Tl) scintillation and a HPGe semiconductor detectors. Nitrogen and chlorine were quantified by analysis of the 10.83-MeV and 6.11-MeV peaks, respectively. Appropriate corrections for the animal body size were determined. The facility described allows the in vivo determination of protein and extracellular space in sets of experimental animals. PMID- 14762645 TI - An automated monitoring system for VOC ozone precursors in ambient air: development, implementation and data analysis. AB - An automated system for the monitoring of volatile organic compound (VOC) ozone precursors in ambient air is described. The measuring technique consists of subambient preconcentration on a cooled trap followed by thermal desorption and GC/FID analysis. First, the technical development, which permits detection limits below 0.05 ppbv to be reached, proceeded in two steps: (1). the determination of optimum sampling parameters (trap composition and conditioning, outlet split, desorption temperature); (2). the development of a reliable calibration method based on a highly accurate standard. Then, a 4-year field application of the hourly measuring chain was carried out at two urban sites. On the one hand, quality control procedures provided the best VOC identification (peak assignment) and quantification (reproducibility, blank system control). On the other hand, the success and performances of the routine experience (88% of the measurements covered more than 40 target compounds) indicated the high quality and suitability of the instrumentation which is actually applied in several French air quality monitoring networks. Finally, an example of data analysis is presented. Data handling identified important organic compound sources other than vehicle exhaust. PMID- 14762644 TI - On-line coupling of microporous membrane liquid-liquid extraction and gas chromatography in the analysis of organic pollutants in water. AB - A method for the determination of hydrophobic pollutants in surface waters was developed. The pretreatment was done with microporous membrane liquid-liquid extraction, the extract was eluted to a sample loop in the large-volume injector valve of the gas chromatograph and the extract was injected on-line to the gas chromatograph. The method was optimised using standard compounds and the linearity, the limits of detection and quantification of the method were studied. The method allowed the determination of hydrophobic pesticides and PAHs at the ng L(-1) level. The RSD values for the repeatability of the method varied from 4.2% to 25.6%, being on average 9.5%. Surface water samples from Finnish lakes and rivers were analysed. PMID- 14762646 TI - Liquid chromatography with triple-quadrupole and quadrupole-time-of-flight mass spectrometry for the determination of micro-constituents - a comparison. AB - The potential of liquid chromatography with triple-quadrupole mass spectrometry (LC-QqQ MS) was compared to that of quadrupole time-of-flight mass spectrometry (LC-Q-ToF MS) for the determination of microconstituents. Three applications were studied: (1) the ng/l quantification of five human drugs in surface water and waste-water effluents; (2) the quantification and confirmation of three corticosteroids in bovine urine at concentrations of 1-100 microg/l, and; (3) the confirmation of nicotine in rat plasma. In all cases, the criteria of the EU Commission Decision 2002/657/EC were followed (for confirmation analysis two MS/MS ions were monitored, and the ratio of their abundances were calculated and compared with those of standards). With both techniques fully satisfactory results were obtained in almost all instances. That is, unequivocal confirmation according to the most stringent EU criteria, those for "illegal compounds", was possible.One main advantage of LC-Q-ToF MS is that for identification and confirmation purposes, full MS/MS spectra are available after a single injection: no second injection, as required with QqQ MS, is needed. As well as the increased efficiency, the enhanced selectivity due to the impressive mass selectivity of LC Q-ToF MS must be emphasized, which allows accurate masses of fragment ions to be calculated. Method characteristics such as linear dynamic range and repeatability were found to be essentially the same for both techniques, but LC-QqQ MS has the advantage that its detection limits are somewhat lower. PMID- 14762647 TI - Air sampling of organophosphate triesters using SPME under non-equilibrium conditions. AB - Solid phase micro-extraction (SPME) was used to collect air samples of semi volatile organophosphate triesters, a group of compounds that are commonly used as flame retardants/plasticisers and have therefore become ubiquitous indoor air pollutants. SPME is a simple sampling technique with several major advantages, including time-efficiency and low solvent consumption. Analyte losses also tend to be relatively low. In quantitative SPME, measurements are normally taken after the analyte has reached partitioning equilibrium between the fibre and the sample matrix. However, equilibrium sampling of semi-volatile compounds in air with SPME often takes several hours. Clearly, time-weighted average (TWA) sampling using SPME under non-equilibrium conditions could be considerably faster. So, in this study, the possibility of sampling organophosphate triesters under non equilibrium conditions was tested. The most important variables proved to be the fibre coating and the air velocity during sampling. The highest uptake rate was obtained with polydimethylsiloxane (PDMS, 100 microm). The rate for this fibre was 150-fold higher than obtained with PDMS/DVB and Carbowax/DVB, both 65 microm. Contrary to theoretical expectations, the uptake rate appeared to be constant for all tested air velocities over the fibre surface >7 cm/s. These findings suggest that the uptake rate for non-equilibrium SPME sampling is independent of the sampling flow above this flow rate, which would considerably enhance the robustness and flexibility of the method. Applying this method for TWA sampling, with sampling periods of 1 h, detection limits lower than 2 ng/m(3) for individual organophosphate esters were obtained. PMID- 14762648 TI - Synthetic polymers adsorbing bisphenol A and its analogues prepared by covalent molecular imprinting using bisphenol A dimethacrylate as a template molecule. AB - Synthetic polymers which can adsorb bisphenol A (BPA) and related compounds were prepared by a covalent molecular imprinting technique. BPA dimethacrylate, used as template molecule, was polymerized with a crosslinker, triethylene glycol dimethacrylate (TEGDMA) or trimethylol propane trimethacrylate (TRIM). After the polymerization treatment with dilute NaOH was used to cleave BPA from the polymers. For high recovery of BPA with low polymer matrix degradation, the hydrolysis conditions were determined to be treatment with 1.0 mol L(-1) NaOH for 48 h. The binding sites generated by the hydrolysis were evaluated by determination of the retentivity of BPA, BPA analogues, and other endocrine disruptors. The polymers strongly adsorbed compounds with two hydroxyl groups at the 4,4'-positions. Generally the TEGDMA-based polymers had stronger affinity than the TRIM-based polymers, although the TRIM-based polymer adsorbed steroidal hormones with two hydroxyl groups, for example 17 alpha-estradiol and 17 beta estradiol, more strongly than the TEGDMA-based polymer, meaning that the crosslinkers affected the properties of the binding sites and, depending upon the target molecules, suitable crosslinkers should be chosen in this system. PMID- 14762649 TI - Comparison of different extraction techniques for the determination of chlorinated pesticides in animal feed. AB - The performances of Soxhlet extraction, dive-in Soxhlet extraction, microwave assisted extraction (MAE), accelerated solvent extraction (ASE), fluidized-bed extraction (FBE), and ultrasonic extraction (UE) for the analysis of organochlorine pesticides in animal feed have been investigated. ASE and MAE provided significantly better extraction efficiency than Soxhlet extraction. The concentrations were 126.7 and 114.8%, respectively, of the values obtained by classical Soxhlet extraction, whereas the results from FBE and dive-in Soxhlet were comparable with those from the standard Soxhlet procedure. The reproducibility of FBE was the best, with RSDs ranging from 0.3 to 3.9%. Under the investigated operation conditions UE was not efficient, with the recoveries of target compounds being about 50% less than Soxhlet. Additionally, the performances of Soxhlet, dive-in Soxhlet, MAE, ASE and FBE were validated by determination of the certified reference material BCR-115. The results from the extraction techniques were in good agreement with the certified values. PMID- 14762652 TI - Oral glucocorticoid use is associated with an increased risk of fracture. AB - Oral glucocorticoids (GCs) are widely used and despite their adverse effects on bone mineral density, the risk of sustaining osteoporotic fractures is not well addressed. The objective of this retrospective, cohort study was to assess fracture risk in patients exposed to oral GCs. Patients from an administrative claims database who were prescribed oral GCs and were enrolled 1 year before and 1 year after the initial oral GC claim were matched with a comparison population on age, sex, and date of first claim. Measurements of exposure included amount, duration, and pattern of oral GC use. The osteoporosis-related risk of fracture was based on the ratio of hazard functions estimated using a Cox proportional hazards model. The adjusted relative risk (RR) estimates (and 95% CI) for fractures were hip 1.87 (95% CI, 1.2 to 2.9), vertebral 2.92 (95% CI, 2.0 to 4.3), wrist/forearm 1.03 (95% CI, 0.8 to 1.4), nonvertebral 1.68 (95% CI, 1.5 to 1.9), any fracture 1.75 (95% CI, 1.6 to 1.9). A dose dependence of fracture risk was observed for hip, vertebral, nonvertebral, and any fractures. Long duration and continuous pattern of GC use demonstrated a significant 5-fold increased risk of hip and 5.9-fold increased risk of vertebral fracture. The combined effect of higher dose, longer duration, and continuous pattern further increased RR estimates to 7-fold for hip and 17-fold for vertebral fractures. This study confirms previous observations that suggest oral GCs have a rapid deleterious effect on trabecular-rich bone. The emerging relationship between amount, duration, and pattern of oral GC exposure and fracture risk should be considered in clinical practice and managed care settings to avoid the debilitating effects of fractures in patients. PMID- 14762653 TI - Evaluation of Medical Outcomes Study Short Form-36 Taiwan version in assessing elderly patients with hip fracture. AB - The Medical Outcomes Study Short Form-36 (SF-36) is a widely used measure of generic health related quality of life. The purpose of this study is to establish the validity and reliability of the SF-36, Taiwan Version, when applied to a sample of elderly patients with hip fracture in Taiwan. Data from two samples were used, the first sample (n = 87) from a prospective descriptive study for testing psychometric scaling assumptions, scale responsiveness and criterion validity, and the second sample (n = 69) from a clinical trial for examining the validity of the differences in the group. The SF-36 Taiwan version demonstrated good evidence of supporting the scaling assumption. Cronbach's alpha coefficients above 0.70 for all scales support the internal consistency. The Physical Function (PF) scale had an effect size of 0.88 from months 1 to 3, and 0.59 from months 3 to 6 after discharge, which appears to have the best responsiveness to clinical changes. Notable floor and ceiling effects (> 15%) for Role Emotion (RE), Role Physical (RP) and PF scales were found. High correlation of 0.62 between the PF and measures of activities of daily living (ADLs), and between RP and instrumental activities of daily living (IADLs) (0.63) supports the construct validity. Significantly higher performance in most SF-36 scales in elders without risk for depression than those who were at risk supported the validity of the group differences. In its current form, the SF-36 Taiwan version demonstrated good reliability and validity as applied to patients with hip fracture. PMID- 14762654 TI - Glucagon-like peptide-1 regulates proliferation and apoptosis via activation of protein kinase B in pancreatic INS-1 beta cells. AB - AIMS/HYPOTHESIS: The incretin hormone glucagon-like peptide-1 augments islet cell mass in vivo by increasing proliferation and decreasing apoptosis of the beta cells. However, the signalling pathways that mediate these effects are mostly unknown. Using a clonal rat pancreatic beta cell line (INS-1), we examined the role of protein kinase B in mediating beta-cell growth and survival stimulated by glucagon-like peptide-1. METHODS: Immunoblot analysis was used to detect active (phospho-) and total protein kinase B. Proliferation was assessed using (3)H thymidine incorporation, while apoptosis was quantitated using 4'-6-diamidino-2 phenylindole staining and APO percentage apoptosis assay. Kinase-dead and wild type protein kinase B was introduced into cells using adenoviral vectors. RESULTS: Glucagon-like peptide-1 rapidly activated protein kinase B in INS-1 cells (by 2.7+/-0.7-fold, p<0.05). This effect was completely abrogated by inhibition, with wortmannin, of the upstream activator of protein kinase B, phosphatidylinositol-3-kinase. Glucagon-like peptide-1 also stimulated INS-1 cell proliferation in a dose-dependent manner (by 1.8+/-0.5-fold at 10(-7) mol/l, p<0.01), and inhibited staurosporine-induced apoptosis (by 69+/-12%, p<0.05). Both of these effects were also prevented by wortmannin treatment. Ablation of protein kinase B by adenovirus-mediated overexpression of the kinase-dead form of protein kinase Balpha prevented protein kinase B phosphorylation and completely abrogated both cellular proliferation ( p<0.05) and protection from drug-induced cellular death ( p<0.01) induced by glucagon-like peptide-1. CONCLUSIONS/INTERPRETATION: These results identify protein kinase B as an essential mediator linking the glucagon-like peptide-1 signal to the intracellular machinery that modulates beta-cell growth and survival. PMID- 14762655 TI - Effects of a long-term treatment with raloxifene on insulin sensitivity in postmenopausal women. AB - AIMS/HYPOTHESIS: Our aim was to investigate the effect of long-term administration of raloxifene, a selective estrogen receptor modulator, on insulin sensitivity, glucose tolerance and plasma lipid concentrations in a group of postmenopausal women. METHODS: A total of 24 women with postmenopausal osteoporosis were consecutively enrolled and randomly assigned to take raloxifene, 60 mg/day for 12 months or placebo. At baseline and after 6 and 12 months, in each subject insulin sensitivity (M-index) was assessed by means of an euglycaemic hyperinsulinaemic clamp. Plasma concentrations of total cholesterol, triglycerides and HDL-cholesterol were also measured and glucose tolerance was evaluated. RESULTS: In the raloxifene-treated group, the M index decreased after 6 and 12 months with respect to the placebo group (-21%, p=0.042 and -23%, p=0.018, respectively). Neither fasting plasma glucose nor glucose tolerance changed in the raloxifene-treated group, compared to the placebo group. Low density lipoprotein cholesterol concentrations decreased at 12 months (-13%, p=0.047). CONCLUSION/INTERPRETATION: A long-term treatment with raloxifene in osteoporotic, otherwise healthy post-menopausal women can reduce insulin sensitivity without affecting glucose tolerance. PMID- 14762656 TI - Insulin enhances vascular cell adhesion molecule-1 expression in human cultured endothelial cells through a pro-atherogenic pathway mediated by p38 mitogen activated protein-kinase. AB - AIMS/HYPOTHESIS: Although hyperinsulinaemia in Type 2 diabetes in states of insulin resistance is a risk factor for atherosclerotic vascular disease, underlying mechanisms are poorly understood. We tested the hypothesis that insulin increases monocyte-endothelial interactions, which are implicated in atherosclerosis. METHODS: We treated human umbilical vein endothelial cells with insulin (10(-10) to 10(-7) mol/l) for 0 to 24 h. To dissect potentially implicated signal transduction pathways, we treated endothelial cells with known pharmacological inhibitors of two distinct insulin signalling pathways: the phosphatidylinositol-3'-kinase (PI3'-kinase) inhibitor wortmannin (3 x 10(-8) to 10(-6) mol/l), involved in insulin-induced endothelial nitric oxide synthase stimulation, and the p38 mitogen-activated protein (p38MAP) kinase inhibitor SB 203580 (10(-7) to 2 x 10(-6) mol/l). We measured adhesion molecule expression by cell surface enzyme immunoassays and U937 monocytoid cell adhesion in rotational adhesion assays. RESULTS: At pathophysiological concentrations (10(-9) to 10(-7) mol/l), insulin concentration-dependently induced vascular cell adhesion molecule (VCAM)-1 (average increase: 1.8-fold) peaking at 16 h. By contrast, the expression of intercellular adhesion molecule-1 and E-selectin were unchanged. The effect on VCAM-1 was paralleled by increased U937 cell adhesion. In the absence of cytotoxicity, wortmannin significantly potentiated the effect of insulin alone on VCAM-1 surface expression and monocytoid cell adhesion, whereas SB-203580 (10(-6) mol/l) completely abolished such effects. CONCLUSIONS/INTERPRETATION: These observations indicate that insulin promotes VCAM-1 expression in endothelial cells through a p38MAP-kinase pathway, amplified by the PI3'-kinase blockage. This could contribute to explaining the increased atherosclerosis occurring in subjects with hyperinsulinaemia, or in states of insulin resistance, which feature a defective PI3'-kinase pathway. PMID- 14762658 TI - [Participation of radiotherapy in interdisciplinary palliative care units- challenge and chance]. AB - BACKGROUND: In Germany, a sufficient system of palliative care does not exist. Possibilities for participation of radiooncologists in the further development of this promising part of medical action are reported. MATERIAL AND METHODS: Experiences from interdisciplinary work in the field of palliative care are described. This experience is communicated for use in the actual discussion about the future of palliative care in Germany, especially in the field of radiooncology. RESULTS: A palliative care unit can only work in a team of different professions, which means different physicians, but also nurses, social workers, psychologists or pastors. A palliative care unit will benefit from working with radiooncologists as well as radiooncologists will do from working in the field of palliative care. CONCLUSION: In times of growing interest in and need for palliative care, radiooncologists should actively participate in the development of palliative care units in Germany. The aim of this participation should be to reasonably arrange the treatment of incurably ill patients with the chances of modern radiotherapy. Another aim should be to improve the treatment of "classic" radiation oncology patients by ideas of palliative care. The further development of palliative care in Germany should not take place without the participation of radiooncologists. This will meet the interests of palliative care and radiotherapy and-most importantly-the patients' interests. PMID- 14762657 TI - The epidemiology of Type 1 diabetes mellitus is not the same in young adults as in children. AB - AIMS/HYPOTHESIS: This prospective study examined the epidemiology of Type 1 diabetes in young adults in Europe. METHODS: We ascertained incident cases of Type 1 diabetes in the 15 to 29 years (both inclusive) age group throughout Europe over a period of 2 years. Diabetes registries in nine countries, in which incidence rates for Type 1 diabetes in the 0 to 14 age group were available, took part. Incidence rates were estimated per 100000 person years and standardised for sex and age. Cumulative incidences per 1000 from birth to age 30 were estimated. Heterogeneity between centres was tested with a Poisson regression model. RESULTS: A total of 2112 diabetes cases were ascertained in 1996 and 1997, of which 61.4% were considered to be Type 1 diabetes. Completeness of ascertainment varied from 70 to 90%. Standardised incidence varied from 4.8 per 100000 person years to 13.4 per 100000 person years. The male-female ratio was estimated to be one or more, and in the 25 to 29 age group 1.5 or more in all countries. Cumulative incidences for males and females indicate that the former exceeds the latter from age 24. In the two centres with highest childhood incidence, this applied already from 14 years of age. CONCLUSIONS/INTERPRETATION: The incidence of Type 1 diabetes in adults is lower than in children and the range of incidence is also reduced, with a less than threefold variation in adults, against an eightfold variation in children. There is a male excess in incidence, especially in the age group 25 to 29 years. PMID- 14762659 TI - Impaired quality of life in patients commencing radiotherapy for cancer. AB - BACKGROUND: This study tested a three-item questionnaire to measure global quality of life (QOL) and pain in patients commencing radiotherapy, based on items from the European Organization for Research and Treatment of Cancer (EORTC) QLQ-C30 instrument. PATIENTS AND METHODS: In a pretest, the EORTC QLQ-C30 and the three-item short questionnaire were administered to 100 patients, yielding similar global QOL and pain scores. After the pretest, the three-item questionnaire was administered to 1,837 patients prior to first radiotherapy treatment. RESULTS: 254 (13.8%) patients with impaired QOL were identified. These patients had a mean global QOL score of 32.6 compared to 72.4 (p < 0.001) found in patients with satisfactory QOL. Patients with impaired QOL were also more likely < 60 years and treated for lung, gastrointestinal or head and neck cancer or advanced, metastatic cancer. CONCLUSION: This brief questionnaire addresses important aspects of QOL, is feasible to use in a clinical setting and therefore represents a potentially useful tool for detecting those patients who may benefit from further evaluation and/or psychosocial support. PMID- 14762660 TI - Decreased local control following radiation therapy alone in early-stage glottic carcinoma with anterior commissure extension. AB - PURPOSE: To assess the patterns of failure in the treatment of early-stage squamous cell carcinoma of the glottic larynx. PATIENTS AND METHODS: Between 1983 2000, 122 consecutive patients treated for early laryngeal cancer (UICC T1N0 and T2N0) by radical radiation therapy (RT) were retrospectively studied. Male-to female ratio was 106 : 16, and median age 62 years (35-92 years). There were 68 patients with T1a, 18 with T1b, and 36 with T2 tumors. Diagnosis was made by biopsy in 104 patients, and by laser vaporization or stripping in 18. Treatment planning consisted of three-dimensional (3-D) conformal RT in 49 (40%) patients including nine patients irradiated using arytenoid protection. A median dose of 70 Gy (60-74 Gy) was given (2 Gy/fraction) over a median period of 46 days (21-79 days). Median follow-up period was 85 months. RESULTS: The 5-year overall, cancer specific, and disease-free survival amounted to 80%, 94%, and 70%, respectively. 5-year local control was 83%. Median time to local recurrence in 19 patients was 13 months (5-58 months). Salvage treatment consisted of surgery in 17 patients (one patient refused salvage and one was inoperable; total laryngectomy in eleven, and partial laryngectomy or cordectomy in six patients). Six patients died because of laryngeal cancer. Univariate analyses revealed that prognostic factors negatively influencing local control were anterior commissure extension, arytenoid protection, and total RT dose < 66 Gy. Among the factors analyzed, multivariate analysis (Cox model) demonstrated that anterior commissure extension, arytenoid protection, and male gender were the worst independent prognostic factors in terms of local control. CONCLUSION: For early-stage laryngeal cancer, outcome after RT is excellent. In case of anterior commissure extension, surgery or higher RT doses are warranted. Because of a high relapse risk, arytenoid protection should not be attempted. PMID- 14762661 TI - Long-term results after external radiotherapy in age-related macular degeneration. A prospective study. AB - PURPOSE: To prospectively evaluate the short- and long-term efficacy of external radiotherapy (RT) in patients with age-related macular degeneration (AMD) by comparing two different dose schedules. PATIENTS AND METHODS: In this prospective, nonrandomized, comparative study including 80 patients, the efficacy of external RT with a total dose of 14.4 Gy (group A, n = 40) and 25.2 Gy (group B, n = 40) was compared. Patients of group A were irradiated between September 1995 and July 1996, patients of group B between August 1996 and November 1997. 67 patients presented with occult choroidal neovascularization (CNV), 13 with classic subfoveal lesions. Complete ophthalmologic investigation was performed before RT, at intervals of 3 months during the 1st year after RT, and of 6 months thereafter. RESULTS: 12 months after RT, vision deteriorated in 85% (14.4 Gy) and 65% (25.2 Gy) of patients. Central visual field decreased with both dose schedules. There was no morphological benefit in neovascular changes. After 48 months, complete follow-up was possible in 46 patients who showed a significant loss of vision similar to the natural course of AMD. CONCLUSION: External RT of AMD with 14.4 Gy as well as with the escalated dose of 25.2 Gy showed a poor beneficial outcome after 6 and 12 months, respectively. After a follow-up of 4 years, visual outcome in irradiated patients was similar to the natural course of the disease. A conspicuous efficacy of RT in prevention of blindness could not be demonstrated. PMID- 14762662 TI - Proton therapy for head and neck malignancies at Tsukuba. AB - PURPOSE: To evaluate the effectiveness and feasibility of proton therapy for head and neck cancers. PATIENTS AND METHODS: From 1983 to 2000, 33 patients with head and neck malignancies but no history of surgical resection were treated with 250 MeV protons with or without X-ray irradiation. This study retrospectively evaluated local control, survival, and treatment sequelae of these patients. The median total target dose using protons with or without X-rays was 76 Gy (range: 42-99 Gy) and the median proton dose per fraction 2.8 Gy (range: 1.5-6.0 Gy). RESULTS: Overall 5-year survival and local control rates were 44% and 74%, respectively. One (3%) and six patients (18%) suffered from treatment-related acute and late toxicity > grade 3 (RTOG/EORTC acute and late radiation morbidity scoring criteria). One patient with a history of radiotherapy suffered from acute toxicity > grade 3. CONCLUSION: Proton therapy appeared to offer high local control rates with few toxicities relative to conventional radiotherapy. However, late toxicity was seen in areas where large radiation doses had been given. PMID- 14762663 TI - Development and evaluation of a skin organ model for the analysis of radiation effects. AB - BACKGROUND AND PURPOSE: The reaction of tissues to ionizing radiation involves alterations in cell-cell and cell-matrix interactions mediated by cellular adhesion molecules. The aim of this study was to develop and evaluate an artificial skin organ model for the analysis of radiation effects. MATERIAL AND METHODS: A human co-culture system consisting of the spontaneously immortalized keratinocyte cell line HaCaT and primary HDFa fibroblasts embedded into a collagen sponge was established. This skin organ model has been characterized and evaluated for its suitability for radiobiological investigations. For that purpose, expression of beta(1)-integrin following irradiation was compared in the skin organ model and in HaCaT monolayer cells (FACScan and immunohistochemistry). Furthermore, the influence of ionizing radiation on DNA fragmentation was investigated in the skin organ model (TUNEL assay). RESULTS: The novel skin organ model showed characteristics of human skin as demonstrated by cytokeratin and Ki 67 immunoreactivity and by electron microscopy. A single dose of 5 Gy X irradiation induced an upregulation of beta(1)-integrin expression both in the skin organ model and in HaCaT cells. Following irradiation, beta(1)-integrin immunoreactivity was intensified in the upper layers of the epidermis equivalent whereas it was almost absent in the deeper layers. Additionally, irradiation of the skin organ model also caused a marked increase of DNA fragmentation. CONCLUSION: These results demonstrate that the novel skin organ model is suitable to investigate cellular radiation effects under three-dimensional conditions. This allows to investigate radiation effects which cannot be demonstrated in monolayer cell cultures. PMID- 14762664 TI - Increased deposition of von Willebrand factor in the rat heart after local ionizing irradiation. AB - BACKGROUND AND PURPOSE: Von Willebrand factor (vWf), a glycoprotein involved in blood coagulation, is synthesized by endothelial cells. Increased amounts of vWf in blood plasma or tissue samples are indicative of damaged endothelium. In the present study, mRNA expression and localization of vWf were determined in irradiated rat heart tissue. MATERIAL AND METHODS: Sprague-Dawley rats received local heart irradiation with a single dose of 0, 15, or 20 Gy. Hearts were dissected at different time points (up to 16 months) after irradiation. In a second experiment, rats were injected with the radioprotector amifostine (160 mg/kg, i. p.) 15-20 min before irradiation and sacrificed after 6 months. Immunohistochemistry was performed using a polyclonal anti-vWf antibody. Serial sections were subjected to a general rat endothelial cell immunostaining (RECA-1) or a collagen staining (picrosirius red). mRNA expression was determined by using PCR. RESULTS: In control tissue, all endothelial cells lining the lumen of the endocardium and coronary arteries, but not capillary endothelial cells, were stained for vWf. 1 month after irradiation with both 15 and 20 Gy, myocardial capillaries became immunoreactive. From 3 months onward, staining was observed also within the extracellular matrix (ECM) of fibrotic areas. At mRNA level, no changes in vWf could be observed at all time points after irradiation, suggesting that vWf deposition was not due to increased biosynthesis of the protein. In sections of amifostine-treated rat hearts, vWf staining was increased to a lesser extent. CONCLUSION: These dose- and time-dependent increases in deposition of vWf indicate the presence of damaged endothelium in the irradiated rat heart. These increases in vWf accumulation precede development of fibrosis in the subendocardial layer and myocardium of the left ventricles, right ventricles, and atria. PMID- 14762665 TI - Reproducibility of patient positioning for fractionated extracranial stereotactic radiotherapy using a double-vacuum technique. AB - BACKGROUND AND PURPOSE: Precise reproducible patient positioning is a prerequisite for conformal fractionated radiotherapy. A fixation system based on double-vacuum technology is presented which can be used for conventional as well as hypofractionated stereotactic extracranial radiotherapy. MATERIAL AND METHODS: To form the actual vacuum mattress, the patient is pressed into the mattress with a vacuum foil which can also be used for daily repositioning and fixation. A stereotactic frame can be positioned over the region of interest on an indexed base plate. Repositioning accuracy was determined by comparing daily, pretreatment, orthogonal portal images to the respective digitally reconstructed radiographs (DRRs) in ten patients with abdominal and pelvic lesions receiving extracranial fractionated (stereotactic) radiotherapy. The three-dimensional (3 D) vectors and 95% confidence intervals (CI) were calculated from the respective deviations in the three axes. Time required for initial mold production and daily repositioning was also determined. RESULTS: The mean 3-D repositioning error (187 fractions) was 2.5 +/- 1.1 mm. The largest single deviation (10 mm) was observed in a patient treated in prone position. Mold production took an average of 15 min (10-30 min). Repositioning times are not necessarily longer than using no positioning aid at all. CONCLUSION: The presented fixation system allows reliable, flexible and efficient patient positioning for extracranial stereotactic radiotherapy. PMID- 14762666 TI - High-dose-rate interstitial brachytherapy for the treatment of penile carcinoma. AB - BACKGROUND: Interstitial low-dose-rate (LDR) brachytherapy allows conservative treatment of T1-T2 penile carcinoma. High dose-rate (HDR) is often considered to be dangerous for interstitial implants because of a higher risk of complications, but numerous reports suggest that results may be comparable to LDR. Nevertheless, there are no data in the literature available regarding HDR interstitial brachytherapy for carcinoma of the penis. CASE REPORT: A 64-year-old man with T1 N0 M0 epidermoid carcinoma of the glans is reported. Interstitial HDR brachytherapy was performed using the stainless hollow needle technique and a breast template for fixation and good geometry. The dose delivered was 18 x 3 Gy twice daily. RESULTS: After 232 days from brachytherapy, the patient was without any evidence of the tumor, experienced no serious radiation-induced complications, and had a fully functional organ. CONCLUSION: HDR interstitial brachytherapy is feasible in selected case of penis carcinoma, when careful planning and small single fractions are used. PMID- 14762667 TI - Effect of different vibration frequencies on heart rate variability and driving fatigue in healthy drivers. AB - OBJECTIVE: This investigation was to assess the effect of different vibration frequencies on heart rate variability (HRV) and driving fatigue in healthy subjects during simulated driving, by the use of power spectrum analysis and subjective evaluation. MATERIALS AND METHODS: Sixty healthy subjects (29.6+/-3.3 years) were randomly divided into three groups, A, B and C, and the subjects of each group participated in the simulated driving for 90 min with vertical sinusoidal vibration (acceleration 0.05 g) of 1.8 Hz (group A), 6 Hz (group B) and no vibration (group C), respectively. Low-frequency (LF) and high-frequency (HF) components of HRV, reflecting sympathetic and parasympathetic activities, and the LF:HF ratio, indicating sympathovagal balance, were measured throughout all periods. All indices of HRV were calculated in the pre-experiment period, mid experiment period and end-experiment period, and were analyzed by repeated measures analysis of variance. Subjective responses to a questionnaire were obtained after the simulated task for the three groups. RESULTS: Significant differences in all indices of HRV were observed between different experiment periods and between any two groups. The ratings of subjective fatigue exhibited significant differences between any two groups. CONCLUSION: The drivers' fatigue ratings were associated with vibration frequencies in simulated driving. The study quantitatively demonstrated that different effects on autonomic nerve activities were induced by different vibration frequencies. PMID- 14762668 TI - Solid cancer incidence among the Chernobyl emergency workers residing in Russia: estimation of radiation risks. AB - An analysis is presented of solid cancer incidence during 11 years of follow-up (1991-2001) of Chernobyl emergency workers residing in Russia. The analysis is based on data from the cohort of male emergency workers from 6 regions in Russia including 55718 persons with documented external radiation doses in the range of 0.001-0.3 Gy who worked within the 30 -km zone in 1986-1987. The mean age at exposure for these persons was 34.8 years and the mean external radiation dose 0.13 Gy. In the cohort 1370 cases of solid cancer were diagnosed and 3 follow-up periods were considered: 1991-1995, 1996-2001 and 1991-2001. The second follow-up period was chosen to allow for a minimum latency period of 10 years being characteristic of solid cancers. For risk assessment two control groups have been introduced, the first 'external' one representing incidence rates for corresponding ages in Russia in general, the second 'internal' one consisting of emergency workers. The risk estimates were based on spontaneous incidence rates of solid cancer. The estimated standardized incidence ratio (SIR) is in good agreement (95% CI) with that of the control. The values of excess relative risk per unit dose (ERR/Gy) for solid malignant neoplasms have been estimated to be 0.33 (95% CI: -0.39, 1.22) (internal control) for the follow-up period 1991-2001 and 0.19 (95% CI: -0.66, 1.27) for 1996-2001. PMID- 14762669 TI - The content of free amino acids in the stratum corneum is increased in senile xerosis. AB - Xerosis is one of the characteristics of aged skin. Xerosis may be caused by a decrease in the stratum corneum free amino acids which are natural moisturizing factors derived from filaggrin. In aged skin, filaggrin is immunohistochemically decreased compared with the levels in young skin. However, the differences in stratum corneum amino acids between aged and young skin have not been analyzed quantitatively. Therefore, in this study we determined the stratum corneum amino acids per 1000 stratum corneum cells in aged and young skin by high-performance liquid chromatography. The amount of filaggrin mRNA in the epidermis was also compared between aged and young skin using RT-PCR. The total amount of amino acids in the stratum corneum was larger in aged senile xerosis skin than in young skin. Only a few amino acids were found in the stratum corneum of ichthyosis vulgaris patients (control skin). The expression of filaggrin mRNA in aged skin was, however, similar to that in young skin. These findings suggest that the immunohistochemical decrease in filaggrin in aged skin may be caused by promotion of filaggrin proteolysis in the upper layers of the stratum spinulosum. PMID- 14762670 TI - Evaluation of the muscles around the knee in rabbits whose anterior cruciate and/or medial collateral ligaments were dissected. AB - INTRODUCTION: The biological response of the muscles around the knee in chronic ligamentous instability was investigated in an animal study. MATERIALS AND METHODS: There were four groups of 6- to 9-month-old adult New Zealand albino rabbits (2500-3300 g). The animals were divided into groups according to the ligament that was surgically sectioned: group A anterior cruciate ligament (ACL), group B medial collateral ligament (MCL), group C both ACL and MCL, and group D served as the control group undergoing no surgical intervention. Three months after surgery, biopsy specimens of the vastus lateralis, rectus femoris, biceps femoris, extensor digitorum longus, and gastrocnemius muscles of the rabbits were obtained. Electron-microscopic cross-sections of the biopsy specimens were evaluated using the new predetermined atrophy parameters. RESULTS: Atrophy was found in the biopsy specimens of the quadriceps muscles in groups A and C (p<0.005). Unimportant changes were seen in the hamstrings, extensor digitorum longus, and gastrocnemius muscles (p>0.05). Only in the group undergoing MCL dissection were no changes observed in the muscles (p>0.05). CONCLUSION: It is concluded that ACL lesions affect the biomechanics of the knee negatively and this situation causes atrophy, especially in the quadriceps muscle. An MCL lesion alone does not cause an important problem in the surrounding musculature, probably because of its spontaneous healing capacity. New criteria for assessment of atrophy in the muscles employing electron-microscopic evaluation are suggested. PMID- 14762671 TI - Clinical evaluation of ulnar nerve repair at wrist level. AB - INTRODUCTION: In this study, the results of ulnar nerve repair were analyzed. The relation between the functional outcome scores and clinical findings were investigated to find out whether any clinical finding could be predictive of the outcome. MATERIALS AND METHODS: Seventeen patients who underwent ulnar nerve repair formed the study group. Average follow-up lasted 45.5 months (range 39-48 months), and average age of the study group was 31.7 years (range 26-42 years). The same operative technique was applied to all patients by one of the authors (HG). Follow-up checks were done at 3, 6, and 12 months postoperatively. The patients who did not attend the last follow-up were excluded from the study group. The Seddon classification was used as the functional scoring system. Wound healing, Tinel sign, interosseous atrophy, atrophy of the first web space, clawing, and protective sensation were the clinical findings examined at the follow-ups. Wound healing was classified as either normal scar formation or hypertrophic scar-keloid (HsC) formation. RESULTS: Good results in 4 and fair results in 13 were obtained according to the Seddon classification. Statistically, there was no difference between the clinical findings at the 3, 6, and 12 month follow-ups. CONCLUSION: Presence of HsC and clawing can be regarded as a predictive sign for fair results in nerve repairs. PMID- 14762672 TI - Failure of prosencephalic unfolding and neuronal migration in acardia. AB - Acardia is a fatal complication of twin pregnancy. It is caused by a retrograde flow of arterial blood from a "pump" into an acardiac twin through placental arterial and venous connections. The heart function of the recipient twin is either blocked or insufficient to support perfusion of the upper body. Severe developmental anomalies ensue. While most acardiacs are anencepahalic, a few twins with a rudimental heart ("hemicardiac") are able to support a variably complex brain. Here we report three cases of hemicardiac twinning with neuronal migrational defects. The most severe abnormalities affected the supratentorial compartment. They can be conceptually divided in two groups. The first, which we tentatively linked to agenesis of the choroid plexus, can be described as a failure of prosencephalic unfolding. The resulting defects included collapsed neocortex, agenesis of the hippocampi and scrambled basal ganglia and diencephalon. The second group of lesions can be theoretically deduced to a result of disruption of the glia-pial boundary with subsequent formation of leptomeningeal heterotopia and zona cerebrovasculosa. Our observations highlight that even in the milieu of a normal genetic background, severe restriction of brain perfusion could lead to neuronal migration defects. Our data also show that adequate unraveling of the brain architecture is crucially dependent on both the parenchymal vascularization and production of the cerebrospinal fluid by the choroid plexus. PMID- 14762673 TI - Activation of vascular endothelial cells and perivascular cells by systemic inflammation-an immunohistochemical study of postmortem human brain tissues. AB - We investigated postmortem human brain tissues to determine whether systemic inflammation causes activation of vascular endothelial cells and perivascular cells. In some cases, we used serum concentrations of an acute-phase reactant, C reactive protein (CRP), as an index of systemic inflammation. Since the serum concentration of CRP at the agonal stage was available only in a limited number of patients, we estimated the degree of systemic inflammation by the intensity of immunohistochemical staining of the residual blood in brain tissue for CRP. Expressions of intercellular adhesion molecule (ICAM)-1, CD40 and cyclooxygenase (COX)-2 were used as markers for activation of vascular endothelial cells. Activation of perivascular cells was estimated by the occurrence of HLA-DR- and CD68-positive perivascular cells. In cases without brain lesions, activation of vascular endothelial cells and perivascular cells was related to the degree of systemic inflammation. In cases with brain lesions, these cells are often activated even in the absence of systemic inflammation. We suggest that inflammatory stimuli derived from the peripheral blood and the brain parenchyma adjunctly activate vascular cells. Under such circumstances, low-grade inflammation in the pre-existing brain lesions might enhance inflammatory signaling to the brain parenchyma from the periphery. The results of this study could explain the vulnerability of neurological patients to delirium caused by systemic inflammatory conditions. PMID- 14762674 TI - Angiolymphoid hyperplasia with eosinophilia involving the cubital nerve. AB - A tumor involving cubital nerve was resected and studied; it was classified as an angiolymphoid hyperplasia with eosinophilia (ALHE). Immunohistochemical and molecular study was done both to confirm the reactive nature of the process and rule out the presence of clonal T or B cell rearrangement. This lesion has been designated as epitheloid hemangioma [Coindre (1994) Ann Pathol 14:426]. Typically, ALHE occurs in the skin and the subcutaneous tissue, and extracutaneous involvement is rare. No cases of ALHE affecting a nerve have been described, but a case of Kimura's disease, the lesions of which have repeatedly been confused with ALHE, has been reported involving median nerve. PMID- 14762675 TI - Frontotemporal dementia with ubiquitinated neuronal inclusions presenting with primary lateral sclerosis and parkinsonism: clinicopathological report of an autopsy case. AB - We report a case displaying upper motor sign, parkinsonism, and behavioral abnormality, with marked degeneration of the precentral cortex, neostriatum and frontotemporal lobes, as well as ubiquitinated neuronal inclusions. The patient was a 66-year-old male at the time of death. At age 57, he noticed progressive difficulties in speaking and swallowing. At age 60, he was severely anarthric and displayed emotional lability and incontinence. Neurologically, very poor movement of tongue was observed, but without atrophy or fasciculation. Deep tendon reflexes were hyperactive. Grasp reflex and snout reflex were also positive. Needle electromyography revealed no abnormalities. A diagnosis of primary lateral sclerosis and character change was made. At age 62, he developed bradykinesia and rigidity of the neck and all extremities. Treatment with carbidopa-levodopa was initiated, but resulted in minimal improvement. At age 65, he was bed-ridden, and had repeated occurrences of aspiration pneumonia; he died of pneumonia. Neuropathological examination revealed marked atrophy of the frontal and temporal lobes with Betz cells completely absent and moderate atrophy of the neostriatum. The spinal cord and nerve roots appeared normal. Immunohistochemically, ubiquitin positive but tau-negative intraneuronal inclusions were found in the frontal and temporal cortices, including the precentral cortex and the hippocampal dentate gyrus, and the neostriatum. This case could be included with inclusion-associated disorders such as frontotemporal dementia or amyotrophic lateral sclerosis with dementia, and furthermore, predominant upper motor sign and parkinsonism could represent phenotypes of clinical manifestations with such inclusions. PMID- 14762676 TI - Expression of ubiquitin-binding protein p62 in ubiquitin-immunoreactive intraneuronal inclusions in amyotrophic lateral sclerosis with dementia: analysis of five autopsy cases with broad clinicopathological spectrum. AB - Amyotrophic lateral sclerosis with dementia (ALSD), corresponding to the motor neuron disease type of frontotemporal dementia, is neuropathologically characterized by depletion of the motor neurons, degeneration of the extra-motor cerebral cortices and formation of ubiquitin-immunoreactive (not argyrophilic, tau-negative, alpha-synuclein-negative) intraneuronal inclusions. Recently, immunoreactivity for ubiquitin-binding protein p62 has been reported in several ubiquitin-containing intraneuronal or intraglial inclusions (e.g. neurofibrillary tangles, Pick bodies, Lewy bodies, glial cytoplasmic inclusions) in various neurodegenerative diseases. We examined p62 immunoreactivity in ubiquitin immunoreactive intraneuronal inclusions in five ALSD cases with a broad clinicopathological spectrum. p62 immunoreactivity in ubiquitin-immunoreactive intraneuronal inclusions was seen in all cases. The mean proportion of p62 immunoreactive inclusions to the total number of ubiquitin-immunoreactive inclusions (p62/Ub ratio) in the dentate gyrus was 27.5 +/- 16.6% (range 6.3 47.3%). There was no correlation between p62/Ub ratio and the severity of dementia, duration of illness or neuropathological severity. Although the main constituent of these inclusions is unknown, our study suggests that p62 contributes to the formation of the inclusions via the same mechanism as in other previously reported neurodegenerative diseases. Since p62 is believed to have a neuroprotective role, the formation of these inclusions may represent a non harmful, rather protective effect against the neuronal degeneration in ALSD. PMID- 14762677 TI - Simplified placement and management of cutting setons in the treatment of transsphincteric anal fistula: technical note. AB - BACKGROUND: We report one safe and standardized technique of seton placement and management. CONCLUSIONS: A simplified way to manage cutting setons helps to minimize manipulation and may reduce pain. In most patients our technique can be used with no additional anesthesia and in an outpatient setting. PMID- 14762679 TI - History, anatomic forms, and pathogenesis of Chiari I malformations. AB - INTRODUCTION: Chiari I malformations constitute a group of entities of congenital or acquired etiology that have in common descent of the cerebellar tonsils into the cervical spinal canal. In recent years, since the advent of magnetic resonance imaging, an increasing number of asymptomatic, doubtfully symptomatic, and minimally symptomatic patients with Chiari I malformations have been diagnosed. This has resulted in controversy about the multiple therapeutic strategies indicated for these problems. OBJECT: With the intention of updating the knowledge that we currently have on the Chiari I malformation and the related syringomyelia we review the literature and discuss the historical background, pathogenesis, anatomic forms, clinical presentation, and diagnostic procedures of these abnormalities. PMID- 14762680 TI - Management of hydrocephalus in posterior fossa tumors: how, what, when? AB - OBJECT: The aim of this work is to analyze the current management of hydrocephalus associated with posterior fossa (PF) tumors. METHODS: The personal perspectives of experienced pediatric neurosurgeons were presented at a virtual round table. DISCUSSION: Preoperative hydrocephalus has been reported in about 80% of patients with PF tumors and postoperative treatment is required for persistent or progressive hydrocephalus in about 25-30% of the cases. Preoperative management includes external ventricular drainage (EVD), endoscopic third ventriculostomy (ETV), shunt insertion, and no treatment at all, while ETV and ventriculo-peritoneal (V-P) shunt are recommended as treatment after PF craniotomy. CONCLUSION: There is no consensus on the way hydrocephalus should be managed before, during, and after PF surgery. While awaiting prospective multicenter trials of various management schemes the perioperative management of hydrocephalus in the context of posterior fossa tumors should be considered as options. PMID- 14762681 TI - Recurrent spinal cord astrocytoma with intraventricular seeding. AB - PATIENT: We report on an unusual case of a recurrent and progressive spinal pilocytic astrocytoma with metastatic spreading to the hypothalamus in a 14-year old boy. TREATMENT AND RESULTS: The patient underwent resection of an intramedullary atypical pilocytic astrocytoma classified as WHO grade II at the level of Th11/12 in 1997 and received local photon beam irradiation. Three years later, a second operation was necessary for a recurrent tumour at the same level. Seventeen months later, a second recurrent tumour with spinal seeding as well as an intracranial tumour in the third ventricle and hypothalamus was detected. He was shunted for an occlusive hydrocephalus and a stereotactic biopsy of the hypothalamic lesion was performed. The tumour was classified as anaplastic pilocytic astrocytoma (WHO grade III). He received chemotherapy with ifosfamide, cisplatin and etoposide (HIT-GBM-C-protocol), craniospinal radiation, and is still alive 60 months after the first operative intervention without neurological deficits. PMID- 14762683 TI - HRCT in children: technique and indications. AB - High-resolution computed tomography (HRCT) is a very important diagnostic tool, which improves our understanding of many lung diseases in children. However, the technique requires great care in managing the child and attention in using the lowest radiation dose possible. HRCT provides important diagnostic information on pediatric lung disorders for both airway and interstitial lung diseases. In this review we describe in detail the most appropriate technique to be used on children including patient preparation and sedation, discuss indications, and analyze the HRCT appearance of a variety of diseases. PMID- 14762682 TI - Neuromodulation for behavior in the locust frontal ganglion. AB - Neuromodulators orchestrate complex behavioral routines by their multiple and combined effects on the nervous system. In the desert locust, Schistocerca gregaria, frontal ganglion neurons innervate foregut dilator muscles and play a key role in the control of foregut motor patterns. To further investigate the role of the frontal ganglion in locust behavior, we currently focus on the frontal ganglion central pattern generator as a target for neuromodulation. Application of octopamine, a well-studied insect neuromodulator, generated reversible disruption of frontal ganglion rhythmic activity. The threshold for the modulatory effects of octopamine was 10(-6) mol l(-1), and 10(-4) mol l(-1) always abolished the ongoing rhythm. In contrast to this straightforward modulation, allatostatin, previously reported to be a myoinhibitor of insect gut muscles, showed complex, tri-modal, dose-dependent effects on frontal ganglion rhythmic pattern. Using a novel cross-correlation analysis technique, we show that different allatostatin concentrations have very different effects not only on cycle period but also on temporal characteristics of the rhythmic bursts of action potentials. Allatostatin also altered the frontal ganglion rhythm in vivo. The analysis technique we introduce may be instrumental in the study of not fully characterized neural circuits and their modulation. The physiological significance of our results and the role of the modulators in locust behavior are discussed. PMID- 14762684 TI - Plant regeneration from protoplasts isolated from embryogenic calli of the forage legume Astragalus melilotoides Pall. AB - An efficient and reproducible protocol is described for the regeneration of Astragalus melilotoides protoplasts isolated from hypocotyl-derived embryogenic calli. Maximum protoplast yield (11.74 +/- 0.6x10(5)/g FW) and viability (87.07 +/- 2.8%) were achieved using a mixture of 2% (w/v) Cellulase Onozuka R10, 0.5% (w/v) Cellulase Onozuka RS, 0.5% (w/v) Macerozyme R10, 0.5% (w/v) Hemicellulase, and 1% (w/v) Pectinase, all dissolved in a cell protoplast wash (CPW) salt solution with 13% (w/v) sorbitol. First divisions occurred 3-7 days following culture initiation. The highest division frequency (9.86 +/- 0.68%) and plating efficiency (1.68 +/- 0.05%) were obtained in solid-liquid medium (KM8P) supplemented with 1.0 mg/l 2,4-dichlorophenoxyacetic acid, 0.5 mg/l 6 benzylaminopurine (BA), 0.2 mg/l kinetin, 0.2 M glucose, 0.3 M mannitol and 500 mg/l casein hydrolysate. Upon transfer to MS medium with 0.5 mg/l alpha naphthaleneacetic acid and 1-2 mg/l BA, the protoplast-derived calli produced plantlets via somatic embryogenesis (56.3 +/- 4.1%) and organogenesis (21.6 +/- 0.6%). Somatic embryos or adventitious shoots developed into well-rooted plantlets on MS medium without any plant growth regulators or supplemented with 3.0 mg/l indole-3-butyric acid, respectively. About 81% of the regenerants survived in soil, and all were normal with respect to morphology and growth characters. PMID- 14762685 TI - Epigenetic dysregulation of the Jak/STAT pathway by frequent aberrant methylation of SHP1 but not SOCS1 in acute leukaemias. AB - SOCS1 and SHP1 are negative regulators of the Jak/STAT signalling pathway that is implicated in leukaemogenesis. We studied if aberrant methylation of SOCS1 and SHP1 might be involved in the pathogenesis and prognostication of acute leukaemias by methylation-specific polymerase chain reaction (MSP). At diagnosis, methylation of SHP1 occurred more frequently in acute myeloid leukaemia (AML) (n=26, 52%) than acute lymphoblastic leukaemia (ALL) (n=6, 24%) (p=0.02). Methylation of SOCS1 was absent in both AML and ALL patients. SHP1 methylation was not associated with specific clinicopathologic features and had no prognostic impact on AML patients. Frequent methylation of SHP1, but not SOCS1, may be important in the pathogenesis, but not prognosis, of acute leukaemias. PMID- 14762693 TI - Capsular neuronal elements and their relation to pain reduction and functional improvement following total hip replacement. AB - We studied changes of pain intensity and functional impairment in 22 patients with osteoarthrosis undergoing total hip replacement. Using a visual analogue scale, the mean scores for pain and disability before surgery were 71.7 and 70.9 respectively. Both scores showed gradual improvement during a 1-year follow-up period, with more than 90% of the total improvement occurring within the first 3 months. After 1 year, the scores for pain and disability were 11.9 and 4.1 respectively. The hip joint capsule was studied using immunohistochemistry to detect neurofilaments. Neurofilament immunoreactivity was observed in 16/22 cases and was correlated with pain and disability scores. However, there were no correlations between pre- and postoperative pain scores, the score changes, and the quantity of capsular neurofilaments. Thus, other factors than capsular neurofilaments influence the scores of pain and disability in osteoarthritis. PMID- 14762694 TI - Correction of relapsed or neglected clubfoot using a simple Ilizarov frame. AB - We present the results of using a simple Ilizarov fixator frame in treatment of 66 feet in 52 patients (mean age 8.5 years) of 58 relapsed and eight neglected clubfeet with grade III or IV severity with a mean follow-up of 40 (26-58) months. Our frame, in spite of being simple and easy for surgeons and patients to handle, achieved satisfactory correction comparable to the literature. PMID- 14762695 TI - CD44v6: a target for antibody-based cancer therapy. AB - The human CD44 gene encodes type 1 transmembrane glycoproteins involved in cell cell and cell-matrix interactions. The structural heterogeneity of the gene products is caused primarily by alternative splicing of at least 10 out of 20 exons. Certain CD44 variant isoforms, in particular those containing CD44 variant domain 6 (CD44v6), have been implicated in tumourigenesis, tumour cell invasion and metastasis. Here we will give an overview of immunohistochemically determined CD44v6 expression in human malignancies (primary epithelial and nonepithelial tumours as well as metastases) and normal tissues, and review several examples of the clinical use of CD44v6-specific antibodies. In nonmalignant tissues, CD44v6 expression is essentially restricted to a subset of epithelia. Intense and homogeneous expression of CD44v6 was reported for the majority of squamous cell carcinomas and a proportion of adenocarcinomas of differing origin, but was rarely seen in nonepithelial tumours. This expression pattern has made CD44v6 an attractive target for antibody-guided therapy of various types of epithelium derived cancers. PMID- 14762696 TI - Sequence-specific DNA strand cleavage by 111In-labeled peptide nucleic acids. AB - Peptide nucleic acids (PNAs) bind tightly and sequence-specifically to single- and double-stranded nucleic acids, and are hence of interest in the design of gene-targeted radiotherapeutics that could deliver the radiodamage to designated DNA and/or RNA sites. As a first step towards this goal, we developed a procedure for incorporation of Auger electron-emitting radionuclide (indium-111) into PNA oligomers and studied the efficiency of PNA-directed cleavage of single-stranded DNA targets. Accordingly, diethylene triamine penta-acetic acid (DTPA) was conjugated to the lysine-appended mixed-base PNAs and sequence-homologous DNA oligomer with a proper linker for comparative studies. By chelation of PNA-DTPA and DNA-DTPA conjugates with (111)In(3+) in acidic aqueous solutions, (111)In labeled PNA and DNA oligomers were obtained. Targeting of single-stranded DNA with PNA-DTPA-[(111)In] conjugates yielded highly localized DNA strand cleavage; the distribution of breaks along the target DNA strand has two maxima corresponding to both termini of PNA oligomer. After 10-14 days, the overall yield of breaks thus generated within the PNA-targeted DNA by (111)In decay was 5 7% versus < or =2% in the case of control oligonucleotide DNA-DTPA-[(111)In]. The estimated yield of DNA strand breaks per nuclear decay is ~0.1 for the PNA directed delivery of (111)In, which is three times more than for the DNA-directed delivery of this radionuclide. This in vitro study shows that (111)In-labeled PNAs are much more effective than radiolabeled DNA oligonucleotides for site specific damaging of DNA targets. Accordingly, we believe that PNA oligomers are promising radionuclide delivery tools for future antisense/antigene radiotherapy trials. PMID- 14762697 TI - Diagnosis and staging of children's lymphoma using the technetium-labelled somatostatin analogue, 99mTc-depreotide. AB - Somatostatin receptor scintigraphy (SRS) is useful in diagnosing tumours with increased expression of somatostatin receptors. Lymphoma cells are known to express somatostatin receptors; however, the application of indium-labelled analogues in children is limited. The aim of this study was to investigate whether the technetium-labelled somatostatin analogue, depreotide, may be useful in diagnosing and staging malignant lymphoma in children. Fifteen children (mean age 13.8 years) with malignant lymphoma were studied (eight with Hodgkin's lymphoma and seven with non-Hodgkin's lymphoma). All children were investigated for verification of staging established by other modalities. Imaging was performed 3-5 h after administration of 300-550 MBq (99m)Tc-depreotide. All patients underwent whole-body scan and single-photon emission tomography of the chest and/or abdomen. Images were assessed visually. In all patients, foci of increased tracer uptake were found. The neck and thorax were the most frequent lesion localisations. Abdominal lesions were found in four patients, and bone lesions in three. In two patients, diffusely increased uptake was observed throughout the skeleton (verified as representing bone marrow involvement). In 11 patients, the number of abnormal sites detected by SRS was greater than that detected by CT. Based on radionuclide examination, three children were upstaged and none were downstaged. It is concluded that (99m)Tc-depreotide shows increased accumulation in pathological sites in malignant lymphoma in children. SRS with (99m)Tc-depreotide provides a single-day imaging method with high sensitivity in lymphoma and with all the advantages of a technetium-labelled compound. Further studies are required on the specificity and the possible value of (99m)Tc depreotide in the follow-up of these patients. PMID- 14762699 TI - Iodine-131 treatment and high-resolution CT: results in patients with lung metastases from differentiated thyroid carcinoma. AB - Between 1984 and 2002, pulmonary metastases were detected in 42 (4%) out of 1,023 patients with differentiated thyroid carcinoma (DTC) in our department. The age at diagnosis ranged from 6 to 77 years. Lung metastases were diagnosed by both increased thyroglobulin (Tg) levels and positive uptake of iodine-131 on scans, and/or positive radiological findings. The primary tumours were histologically classified as papillary (30 patients), follicular (nine patients) and poorly differentiated (two tall cell, one insular carcinoma). The duration of follow-up ranged from 24 to 228 months. The end-results of the (131)I therapy were evaluated. The treatment of choice was (131)I therapy of metastases after total thyroidectomy plus lymph node dissection (if lymph node metastases were present). Applied single and total (131)I activities were 1.8-10.4 GBq and 5.5-43.7 GBq, respectively. Lung metastases were present at the time of diagnosis in 30 patients and developed during the follow-up period in the remaining 12. Twelve patients with extensive metastases died of thyroid carcinoma and another died due to secondary malignancy (malignant mesothelioma). Ten patients with lung metastases remain completely free of disease and are probably cured, while another seven were stable at the time of study. Three- and five-year survival rates were 86% (36/42) and 76% (32/42), respectively. To define the diagnostic value of high-resolution computed tomography (HRCT) and identify the distinctive features of lung metastases from DTC, 22 patients were further examined with HRCT within 2 weeks of the initial diagnosis of lung metastases and the results were compared with chest X-ray findings. HRCT detected metastases in 10 out of 14 patients with a normal chest X-ray and confirmed metastases in all patients with positive (n=5) and suspicious (n=3) chest X-ray. HRCT did not show any abnormalities in four patients with positive lung uptake on (131)I whole-body images. Stage of disease, existence of distance metastases other than to the lung, and HRCT characteristics were significant prognostic variables. Lung metastases from DTC can be cured with (131)I therapy in a considerable number of patients, especially when they are not associated with other distant metastases; they should therefore be treated at an early stage. HRCT clearly improved diagnostic ability in the evaluation of lung metastases compared with chest X-ray and should be the primary method when radiological correlation is needed. The main, and new, finding of the study is that HRCT not only improves diagnostic ability but is also of prognostic value. PMID- 14762698 TI - Response prediction by FDG-PET after neoadjuvant radiochemotherapy and combined regional hyperthermia of rectal cancer: correlation with endorectal ultrasound and histopathology. AB - Accurate response assessment after neoadjuvant therapy is essential in patients with rectal cancer. The aim of this study was to assess the value of fluorine-18 fluorodeoxyglucose positron emission tomography (FDG-PET) in predicting response of locally advanced rectal cancer to preoperative multimodal treatment. Twenty two consecutive patients with locally advanced (uT3/4) primary rectal cancer were entered in this prospective pilot study. FDG-PET was performed before and after neoadjuvant radiochemotherapy (RCT) with combined regional hyperthermia (RHT). Treatment consisted of external-beam radiotherapy (45 Gy), chemotherapy (folinic acid and 5-fluorouracil) and regional pelvic hyperthermia followed by curative tumour resection 6-8 weeks later. Semi-quantitative measurements (SUV) of tumour FDG uptake were made before and 2-4 weeks after completion of neoadjuvant treatment. Two patients who did not receive post-therapeutic restaging by FDG-PET were excluded from the analysis. Results were correlated with findings on endorectal ultrasound (EUS, n=17 patients) and histopathology. Histopathological evaluation of the resected tumour revealed complete response in one patient, partial response in 12 and stable disease in seven. SUV reduction in tumours was significantly greater in responders than in non-responders [60% (+/-15%) vs 30% (+/-18%), P=0.003, CI=95%). Using a minimum post-therapeutic SUV reduction of 36% to define response, FDG-PET revealed a sensitivity of 100% (EUS: 33%) and a specificity of 86% (EUS: 80%) in response prediction; the corresponding positive and negative predictive values were 93% (EUS: 80%) and 100% (EUS: 33%), respectively. FDG-PET results were statistically significant (P<0.001, CI=95%). FDG-PET has great potential in the assessment of tumour response to neoadjuvant RCT in combination with RHT and is superior to EUS for this purpose. PMID- 14762701 TI - Lincomycin, clindamycin and their applications. AB - Lincomycin and clindamycin are lincosamide antibiotics used in clinical practice. Both antibiotics are bacteriostatic and inhibit protein synthesis in sensitive bacteria. They may even be bactericidal at the higher concentrations that can be reached in vivo. Clindamycin is usually more active than lincomycin in the treatment of bacterial infections, in particular those caused by anaerobic species; and it can also be used for the treatment of important protozoal diseases, e.g. malaria, most effectively in combination with primaquine. Resistance to lincomycin and clindamycin may be caused by methylation of 23S ribosomal RNA, modification of the antibiotics by specific enzymes or active efflux from the periplasmic space. PMID- 14762700 TI - Has PET become an important clinical tool in paediatric imaging? PMID- 14762703 TI - An equation for calculating the volumetric ratios required in a ligation reaction. AB - The ligation of two DNA fragments to create a new plasmid DNA molecule is a key reaction in molecular biology. Where the fragment lengths and concentrations are known, existing equations allow the desired relative molar ratio to be calculated, but this must then be related to the required volumes. Further calculations are then necessary if the maximum available volume is to consist of DNA solutions. The equation presented here allows the simple calculation of volumes of DNA solutions required to obtain a desired molar insert-to-vector ratio, and these can comprise all of the available volume in a ligation if required, thus maximising the yield of the recombinant plasmid. PMID- 14762702 TI - Quinone-respiration improves dechlorination of carbon tetrachloride by anaerobic sludge. AB - The impact of humic acids and the humic model compound, anthraquinone-2,6 disulfonate (AQDS), on the biodegradation of carbon tetrachloride (CT) by anaerobic granular sludge was studied. Addition of both humic acids and AQDS at sub-stoichiometric levels increased the first-order rate of conversion of CT up to 6-fold, leading to an increased production of inorganic chloride, which accounted for 40-50% of the CT initially added. Considerably less dechlorination occurred in sludge incubations lacking humic substances. By comparison, very limited dechlorination occurred in sterile controls with autoclaved sludge. Accumulation of chloroform (1-10%) and dichloromethane (traces) also accounted for the CT converted. The accumulation of a chlorinated ethene, perchloroethylene (up to 9% of added CT), is also reported for the first time as an end-product of CT degradation. A humus-respiring enrichment culture (composed primarily of a Geobacter sp.) derived from the granular sludge also dechlorinated CT, yielding products similar to the AQDS-supplemented granular sludge consortium. The dechlorination of CT by the Geobacter enrichment was dependent on the presence of AQDS or humic acids, which were reduced during the assays. The reduced form of AQDS, anthrahydroquinone-2,6-disulfonate, was shown to cause the chemical reduction of CT when incubated in sterile medium. The results taken as a whole indicate that the formation of reduced humic substances by quinone-respiring microorganisms can contribute to the reductive dechlorination of CT. PMID- 14762704 TI - Current-voltage-time records of ion translocating enzymes. AB - Membrane currents, as non-linear functions of membrane voltage, V, and time, t, can be recorded quickly by triangular V protocols. From the differences, d I(V, t), of these relationships upon addition of a putative substrate of a charge translocating membrane protein, the I(V, t) relationships of the transporter itself can be determined. These relationships likely comprise a steady-state component, I(a)(V), of the active transporter, and a dynamic component, p(a)(V, t), of its V- and time-dependent activity, p(a). Here, the steady-state component is modeled by a central reaction cycle, which senses a fraction delta(tr) of the total V, whereas 1-delta(tr) can be assigned to an inner and outer pore section with delta(i) and delta(o), respectively (delta(i)+delta(tr)+delta(o) = 1). For the enzymatic cycle, fast binding/debinding is assumed, plus V-sensitive and insensitive reaction steps which may become rate limiting for charge translocation. At given substrate concentrations, I(a)(V) is defined by eight independent system parameters, including a coefficient for the barrier shape of charge translocation. In ordinary cases, the behavior of p(a)(V, t) can be described by two rate constants (for activation and inactivation) and their respective V-sensitivity coefficients. Here, the effects of the individual system parameters on I(V, t) from triangular V-clamp experiments are investigated systematically. The results are illustrated by panels of typical curve shapes for non-gated and gated transporters to enable a first classification of mechanisms. We demonstrate that all system parameters can be determined fairly well by fitting the model to "experimental" data of known origin. Applicability of the model to channels, pumps and cotransporters is discussed. PMID- 14762705 TI - Resolving the molecular structure of microtubules under physiological conditions with scanning force microscopy. AB - We have imaged microtubules, essential structural elements of the cytoskeleton in eukaryotic cells, in physiological conditions by scanning force microscopy. We have achieved molecular resolution without the use of cross-linking and chemical fixation methods. With tip forces below 0.3 nN, protofilaments with approximately 6 nm separation could be clearly distinguished. Lattice defects in the microtubule wall were directly visible, including point defects and protofilament separations. Higher tip forces destroyed the top half of the microtubules, revealing the inner surface of the substrate-attached protofilaments. Monomers could be resolved on these inner surfaces. PMID- 14762706 TI - Calculating biological behaviors of epigenetic states in the phage lambda life cycle. AB - The biology and behavior of bacteriophage lambda regulation have been the focus of classical investigations of molecular control of gene expression. Both qualitative and quantitative aspects of this behavior have been systematically characterized experimentally. Complete understanding of the robustness and stability of the genetic circuitry for the lysis-lysogeny switch remains an unsolved puzzle. It is an excellent test case for our understanding of biological behavior of an integrated network based on its physical, chemical, DNA, protein, and functional properties. We have used a new approach to non-linear dynamics to formulate a new mathematical model, performed a theoretical study on the phage lambda life cycle, and solved the crucial part of this puzzle. We find a good quantitative agreement between the theoretical calculation and published experimental observations in the protein number levels, the lysis frequency in the lysogen culture, and the lysogenization frequency for mutants of O(R). We also predict the desired robustness for the lambda genetic switch. We believe that this is the first successful example in the quantitative calculation of robustness and stability of the phage lambda regulatory network, one of the simplest and most well-studied regulatory systems. PMID- 14762707 TI - Carbamoylphosphonate-based matrix metalloproteinase inhibitor metal complexes: solution studies and stability constants. Towards a zinc-selective binding group. AB - Overactive matrix metalloproteinases (MMPs) are associated with a variety of disease states. Therefore, their inhibition is a highly desirable goal. Yet, more than a decade of worldwide activity has not produced even one clinically useful inhibitor. Because of the crucial role of zinc in the activity of the enzyme, the design of inhibitors is usually based upon a so-called zinc binding group (ZBG). Yet, many of the hitherto synthesized potent inhibitors failed clinically, presumably because they bind stronger to metals other than zinc. We have developed in vivo potent inhibitors based on the carbamoylphosphonic group as a putative ZBG. In this paper we report stability constants for Ca(II), Mg(II), Zn(II) and Cu(II) complexes of two potent, in vivo active, MMP inhibitors, cyclopentylcarbamoylphosphonic acid (1) and 2-( N, N dimethylamino)ethylcarbamoylphosphonic acid (2). Precipitation prevented the determination of stability constants for iron(III) complexes of1 and2. For comparison with carbamoylphosphonates1 and2, we synthesized 2-cyclohexyl-1,1 difluoroethylphosphonic acid (3), which does not inhibit MMP, and determined the stability constants of its complexes with Mg(II), Ca(II) and Zn(II). Comparison with the values obtained from the complexes of1 and2 with those from3 indicates participation of the C=O group in the metal binding of the former compounds. The complex stability orders for both1 and2 are Ca(II)8 the dimethylamino group of compound2 can also participate in the binding of the transition metals Cu and Zn. On the other hand, the amino group in carbamoylphosphonic acid2 lowers the stability of the complexes with metals favoring oxygen ligands (Ca, Mg and Fe) and increases the selectivity towards Zn. These results are helpful for rationalizing the results observed on our MMP inhibitors hitherto examined, and are expected to be useful for the design of new selective inhibitors. PMID- 14762708 TI - Dropped head syndrome in mitochondriopathy. AB - In a 63-year-old, 165-cm-tall woman with a history of repeated tick bites, dilative cardiomyopathy, osteoporosis, progressive head ptosis with neck stiffness and cervical pain developed. The family history was positive for thyroid dysfunction and neuromuscular disorders. Neurological examination revealed prominent forward head drop, weak anteflexion and retroflexion, nuchal rigidity, weakness of the shoulder girdle, cogwheel rigidity, and tetraspasticity. The lactate stress test was abnormal. Electromyograms of various muscles were myogenic. Muscle biopsy showed non-specific myogenic abnormalities and generally weak staining for cytochrome oxydase. Mitochondriopathy with multi organ involvement was suspected. The response to anti-Parkinson medication was poor. In conclusion, dropped head syndrome (DHS) may be due to multi-organ mitochondriopathy, manifesting as Parkinsonism, tetraspasticity, dilative cardiomyopathy, osteoporosis, short stature, and myopathy. Anti-Parkinson medication is of limited effect. PMID- 14762709 TI - Development of two bone-derived cell lines from the marine teleost Sparus aurata; evidence for extracellular matrix mineralization and cell-type-specific expression of matrix Gla protein and osteocalcin. AB - A growing interest in the understanding of the ontogeny and mineralization of fish skeleton has emerged from the recent implementation of fish as a vertebrate model, particularly for skeletal development. Whereas several in vivo studies dealing with the regulation of bone formation in fish have been published, in vitro studies have been hampered because of a complete lack of fish-bone-derived cell systems. We describe here the development and the characterization of two new cell lines, designated VSa13 and VSa16, derived from the vertebra of the gilthead sea bream. Both cell types exhibit a spindle-like phenotype and slow growth when cultured in Leibovitz's L-15 medium and a polygonal phenotype and rapid growth in Dulbecco's modified Eagle medium (D-MEM). Scanning electron microscopy and von Kossa staining have revealed that the VSa13 and VSa16 cells can only mineralize their extracellular matrix when cultured in D-MEM under mineralizing conditions, forming calcium-phosphate crystals similar to hydroxyapatite. We have also demonstrated the involvement of alkaline phosphatase, a marker of bone formation in vivo, and Gla proteins (osteocalcin and matrix Gla protein, MGP) in the process of mineralization. Finally, we have shown that VSa13 and VSa16 cell lines express osteocalcin and MGP in a mutually exclusive manner. Thus, both cell lines are capable of mineralizing in vitro and of expressing genes found in chondrocyte and osteoblast cell lineages, emphasizing the suitability of these new cell lines as valuable tools for analyzing the expression and regulation of cartilage- and bone-specific genes. PMID- 14762710 TI - Tyrosine kinase receptor expression in thymomas. AB - PURPOSE: Promising new therapies for neoplasia include tyrosine kinase receptor antagonists. Tyrosine kinase oncogenes present an appealing anti-tumor drug target since they play an integral role in a variety of cellular responses including cell proliferation and differentiation. We previously demonstrated a high rate of epidermal growth factor receptor (EGFR) expression in advanced-stage thymic epithelial tumors. More recently, we have examined c-KIT (CD117) expression in a similar series of tumors. METHODS: Tumor from 35 patients seen at our institution for treatment of advanced-stage thymoma was available. Twenty thymomas and 15 thymic carcinomas were assessed for c-KIT expression. Tissue sections of tumor were stained immunohistochemically with anti-c-KIT (Oncogene). Either cytoplasmic or membrane staining was considered positive. Appropriate controls were performed. Positive staining for c-KIT was present in 12 tumors (11 thymic carcinomas and 1 thymoma). RESULTS: In distinction to EGFR, c-KIT is expressed more commonly in thymic carcinomas (73% of carcinomas) than in thymoma (5% of thymomas). CONCLUSIONS: An EGFR negative/c-KIT positive staining pattern is typical of thymic carcinoma, whereas thymomas are generally EGFR positive/c KIT negative. Possible therapeutic implications of these observations remain to be determined. PMID- 14762711 TI - The use of a hand-held metal detector for localisation of ingested metallic foreign bodies - a critical investigation. AB - Ingested metallic foreign bodies (MFBs) are usually diagnosed by taking X-ray films of the neck, chest and/or abdomen. This study evaluates the use of a hand held metal detector (HHMD) for the diagnosis and localisation of MFBs. In a prospective study, 53 consecutive paediatric patients with history of a swallowed MFB were examined with X-rays and HHMD. In 47 children, the MFB could be verified radiologically. Coins were most frequently swallowed. The HHMD could detect and locate all coins but only 47% of other MFBs. There were no false-positive results. A HHMD is an effective tool for screening the location of suspected ingested coins. This method is easy, inexpensive and free of radiation. Very small MFBs cannot be reliably detected. CONCLUSION: If an innocuous metallic foreign body is clearly identified with a hand-held metal detector in the stomach or lower gastrointestinal tract of an asymptomatic child, additional radiological confirmation is not required. PMID- 14762712 TI - Severe encephalopathy with epilepsy in an infant caused by subclinical maternal pernicious anaemia: case report and review of the literature. AB - Vitamin B(12) deficiency is one of the major causes of megaloblastic anaemia with or without neurological symptoms. We report on a patient manifesting acute encephalopathy, epilepsy, microcephaly and megaloblastic anaemia at the age of 4 months. Vitamin B(12) deficiency in the patient was due to subclinical pernicious anaemia of the mother who exhibited neither haematological nor neurological symptoms. Mother and child both had elevated methylmalonic acid in their urine which is a sensitive parameter of vitamin B(12) deficiency. Vitamin B(12) therapy resulted in arrest of convulsions within 24 h. There were no further seizures although the patient showed moderate mental retardation at the age of 7 years but a normal head circumference. Long-term MRI follow-up, performed at the age of 7 years, showed moderate enlargement of the ventricles with reduction of myelin and hypoplasia of the corpus callosum. CONCLUSION: Vitamin B(12) deficiency due to maternal pernicious anaemia should always be considered in the differential diagnosis of neurological symptoms in infants and especially in combination with megaloblastic anaemia. Since the age of onset and the duration of neurological symptoms may contribute to the development of long-term symptoms, early diagnosis and treatment is important for vitamin B(12) deficient children. PMID- 14762713 TI - Regeneration in chronic sialadenitis: an analysis of proliferation and apoptosis based on double immunohistochemical labelling. AB - The understanding of regeneration in salivary glands as a finely tuned balance of cellular proliferation, differentiation and apoptosis has been limited by the difficulty of identifying proliferating cells. This has been overcome in the present investigation by double immunohistochemical labelling for the proliferation-associated antigen Ki67 and for different cell-type-specific antigens applied to 8 specimens of normal parotids and 16 specimens of chronic parotid sialadenitis with particular reference to acini and intercalated ducts. In comparison with low baseline rates of proliferation in normal parotids, proliferative indices were significantly increased in chronic sialadenitis in mature acinar cells, intercalated ductal cells and myoepithelial cells without evidence of proliferation by an additional population of cells. In accordance with findings in glands of experimental animals, the present data do not support the previously postulated concept of regeneration of acini and intercalated ducts by a hypothetical population of uncommitted ductal stem cells. The demonstration of a profound capacity for intrinsic glandular regeneration from differentiated cells represents a biological basis for the good results obtained from conservative therapy of chronic sialadenitis and offers hope for novel therapies designed to reconstitute impaired salivary flow. PMID- 14762714 TI - Nitric oxide generation and poly(ADP ribose) polymerase activation precede beta cell death in rats with a single high-dose injection of streptozotocin. AB - Streptozotocin (STZ) is widely used for the induction of diabetes in animals by causing destruction of pancreatic beta cells. This experiment was designed to elucidate the sequential process of beta-cell destruction in rats with a single high-dose injection of STZ. At 0, 2, 5, 8 and 24 h after injection, rats were perfused with Krebs-Ringer buffer with dichlorofluorescein diacetate (DCF-DA), a marker for free radicals, and the pancreata were pathologically analyzed. Injection of STZ rapidly elicited an increase in fluorescence of DCF-DA in beta cells at 2 h after the injection. The fluorescence was diminished by carboxy PTIO, a specific scavenger of nitric oxide (NO), but not by L-NAME, an inhibitor of NO synthase. During this process, an inducible form of NO synthase was not detected. Thereafter, upregulated expression of poly(ADP ribose) polymerase (PARP) and massive beta-cell death were detected at 5-8 h after injection. Migration of macrophages into the islet was conspicuous at 24 h, clearing up the debris of destroyed beta cells. Nicotinamide, a PARP inhibitor, significantly inhibited beta-cell death without apparent suppression of NO generation at 2 h. The current study documented serial processes of STZ-induced beta-cell death, starting with NO generation and PARP activation followed by a clearance with macrophages, where the activation of PARP plays a central role in beta-cell death. PMID- 14762715 TI - Melanotic oncocytic metaplasia of the nasopharynx: a report of seven cases and review of the literature. AB - We describe seven cases of melanotic oncocytic metaplasia of the nasopharynx and review five other cases in the literature. It is usually a small, brown to black lesion that occurs around the Eustachian tube opening, where abundant seromucinous glands and lymphoid tissue are present. Multiple or bilateral lesions are sometimes seen. All 12 reported cases are of Asian origin. Melanotic oncocytic metaplasia occurs predominantly in men (male:female=11:1), with a mean age of 68 years. Simple excisional biopsy appears to be curative. Microscopically, melanotic oncocytic metaplasia is a combination of oncocytic metaplasia of the epithelium of the gland and melanin pigmentation in its cytoplasm. Fontana-Masson staining and immunohistochemical staining of S-100 protein revealed numerous melanocytes with conspicuous dendrites in the glands and stroma, which probably transfer melanin to adjacent glands. The exact pathogenesis of melanotic oncocytic metaplasia is unknown, but we postulate that the lesion could be related to the oncocytic metaplasia of the seromucinous glands around the Eustachian tube, which is followed by the local production and/or acquisition of the melanin pigment, under the influence of certain neuropeptides in the vicinity. The recognition of melanotic oncocytic metaplasia is of clinical importance, as it may be misdiagnosed as a malignancy to the unwary. PMID- 14762716 TI - What's at the top in the top-down control of action? Script-sharing and 'top-top' control of action in cognitive experiments. AB - The distinction between bottom-up and top-down control of action has been central in cognitive psychology, and, subsequently, in functional neuroimaging. While the model has proven successful in describing central mechanisms in cognitive experiments, it has serious shortcomings in explaining how top-down control is established. In particular, questions as to what is at the top in top-down control lead us to a controlling homunculus located in a mythical brain region with outputs and no inputs. Based on a discussion of recent brain imaging experiments, we argue for the need to factor the interaction between the experimenter and the experimental participant into a realistic understanding of top-down control. We suggest these interactions involve a 'sharing of scripts' for perception and action that may be described as 'top-top processes.' We thereby expand the understanding of the homunculus to include elements of social cognition. This conceptual reconfiguration may grant some sort of asylum for a- not very omnipotent--homunculus. PMID- 14762720 TI - Simulation and parameter estimation of dynamics of synaptic depression. AB - Synaptic release was simulated using a Simulink sequential storage model with three vesicular pools. Modeling was modular and easily extendable to the systems with greater number of vesicular pools, parallel input, or time-varying parameters. Given an input (short or long tetanic trains, patterned or random stimulation) and the storage model, the vesicular release, the replenishment of various vesicular pools, and the vesicular content of all pools could be simulated for the time-invariant and time-varying storage systems. From the input stimuli and either a noiseless or a noisy output, the parameters of such storage systems could also be estimated using the optimization technique that minimizes in the least square sense the error between the observed release and the predicted release. All parameters of the storage model could be evaluated with sufficiently long input-output data pairs. Not surprisingly, the parameters characterizing the processes near the release locus, such as the fractional release and the size of the immediately available pool and its coupling to the small store, as well as the state variables associated with the immediately available pool, such as its vesicular content and replenishment, could be determined with fewer stimuli. The possibility of estimating parameters with random inputs extends the applicability of the method to in vivo synapses with the physiological inputs. The parameter estimation was also possible under the time-variant, but slowly changing, conditions as well as for open systems that are part of larger vesicular storage systems but whose parameters can either not be reliably determined or are of no interest. The quality of parameter estimation was monitored continuously by comparing the observed and predicted output and/or estimated parameters with the true values. Finally, the method was tested experimentally using the rat phrenic-diaphragm neuromuscular junction. PMID- 14762718 TI - Hypoxia up-regulates triosephosphate isomerase expression via a HIF-dependent pathway. AB - The glycolytic enzyme triosephosphate isomerase (TPI) catalyses the reversible conversion of dihydroxyacetone phosphate into glyceraldehyde-3-phosphate. We report here that the expression of TPI at both the mRNA and protein levels is increased by hypoxia in vivo and in vitro. The temporal pattern of hypoxic TPI induction is very similar to that of genes triggered by the hypoxia-inducible transcription factor (HIF) and is mimicked characteristically by cobalt and by deferoxamine, but is absent in cells with a defective aryl hydrocarbon receptor nuclear translocator (ARNT, here HIF-1beta) and in cells lacking HIF-1alpha protein. We conclude from these findings that the expression of TPI is regulated via the HIF pathway and thus belongs to the family of classic oxygen-regulated genes. The physiological meaning of an increased expression of TPI in hypoxygenated tissues is probably to increase the flow of triosephosphates through the glycolytic cascade thus leading to an increase of anaerobic energy generation. PMID- 14762721 TI - Information encoding in the inferior temporal visual cortex: contributions of the firing rates and the correlations between the firing of neurons. AB - The encoding of information by populations of neurons in the macaque inferior temporal cortex was analyzed using quantitative information-theoretic approaches. It was shown that almost all the information about which of 20 stimuli had been shown in a visual fixation task was present in the number of spikes emitted by each neuron, with stimulus-dependent cross-correlation effects adding for most sets of simultaneously recorded neurons almost no additional information. It was also found that the redundancy between the simultaneously recorded neurons was low, approximately 4% to 10%. Consistent with this, a decoding procedure applied to a population of neurons showed that the information increases approximately linearly with the number of cells in the population. PMID- 14762722 TI - Quantum Zeno features of bistable perception. AB - A generalized quantum theoretical framework, not restricted to the validity domain of standard quantum physics, is used to model the dynamics of the bistable perception of ambiguous visual stimuli such as the Necker cube. The central idea is to treat the perception process in terms of the evolution of an unstable two state system. This gives rise to a "Necker-Zeno" effect, in analogy to the quantum Zeno effect. A quantitative relation between the involved time scales is theoretically derived. This relation is found to be satisfied by empirically obtained cognitive time scales relevant for bistable perception. PMID- 14762723 TI - The world from a cat's perspective--statistics of natural videos. AB - The mammalian visual system is one of the most intensively investigated sensory systems. However, our knowledge of the typical input it is operating on is surprisingly limited. To address this issue, we seek to learn about the natural visual environment and the world as seen by a cat. With a CCD camera attached to their head, cats explore several outdoor environments and videos of natural stimuli are recorded from the animals' perspective. The statistical analysis of these videos reveals several remarkable properties. First, we find an anisotropy of oriented contours with an enhanced occurrence of horizontal orientations, earlier described in the "oblique effect" as a predominance of the two cardinal orientations. Second, contrast is not elevated in the center of the images, suggesting different mechanisms of fixation point selection as compared to humans. Third, analyzing a sequence of images we find that the precise position of contours varies faster than their orientation. Finally, collinear contours prevail over parallel shifted contours, matching recent physiological and anatomical results. These findings demonstrate the rich structure of natural visual stimuli and its direct relation to extensively studied anatomical and physiological issues. PMID- 14762724 TI - Stability in legged locomotion. AB - Stability is a key element in a gait synthesis. Static stability margins are widely adopted in crawlers, while no similar approach has been developed for dynamically stable systems. Utilizing an analytical approach, we developed a set of easy-to-calculate stability indices to describe instantaneous static and dynamic (In)stability for a certain group of walking systems. The analysis is based on a thorough analysis of the interaction between ground reaction forces and the walking system. The indices are applicable to walking systems regardless of the number of legs or mechanical/biological design. We show that static and dynamic stability are independent of each other. We suggest a possible categorization of gait modes based on stability. Stability characteristics are analyzed in a healthy and highly pathological human gait. Finally, we discuss the applicability of the proposed methods. PMID- 14762725 TI - Intrinsic bursting activity in the pre-Botzinger complex: role of persistent sodium and potassium currents. AB - Computational models of single pacemaker neuron and neural population in the pre Botzinger Complex (pBC) were developed based on the previous models by Butera et al. (1999a,b). Our modeling study focused on the conditions that could define endogenous bursting vs. tonic activity in single pacemaker neurons and population bursting vs. asynchronous firing in populations of pacemaker neurons. We show that both bursting activity in single pacemaker neurons and population bursting activity may be released or suppressed depending on the expression of persistent sodium ( I(Na) P) and delayed-rectifier potassium ( I(K)) currents. Specifically, a transition from asynchronous firing to population bursting could be induced by a reduction of I(K) via a direct suppression of the potassium conductance or through an elevation of extracellular potassium concentration. Similar population bursting activity could be triggered by an augmentation of I(Na) P. These findings are discussed in the context of the possible role of population bursting activity in the pBC in the respiratory rhythm generation in vivo vs. in vitro and during normal breathing in vivo vs. gasping. PMID- 14762726 TI - [Homocysteine: vascular risk factor?]. PMID- 14762727 TI - [Antioxidant enzymes in acute cerebral infarction]. AB - BACKGROUND AND PURPOSE: Oxidative modification of DNA, protein and lipids by reactive oxygen peroxides plays an important role in acute cerebral infarction. The biological antioxidant defence system against oxidative stress is an integrated array of enzymes such as superoxide dismutase (SOD), catalase (CAT) and glutathion peroxidase (GPx). The purpose of this work was: 1. To investigate the evolution of SOD and CAT activities as possible markers of oxidative activity in patients with acute cerebral infarction. 2. To establish the relationship between the type of stroke and the activity of these two enzymes. 3. To analyze the relationship between the degree of neurological deficits of patients with stroke and SOD and CAT activities. PATIENTS AND METHODS: This is a prospective study comprising 45 patients with acute cerebral infarction (24 females and 21 males), aged from 48 to 96. The infarcts were classified in atherotrombotics and cardioembolics. SOD and CAT activities were measured on days 1, 3, 6 and 15. RESULTS: Activities of SOD and CAT in patients on admission were lower than those of controls and showed a gradual increase over time. The activity mean of SOD was significantly higher in cardioembolic infarctions than in atherotrombotic infarctions (p < 0.05). SOD activity was significantly lower for Mathew score > 60, while CAT activity was higher. CONCLUSIONS: Our findings suggest that there is a progressive increase in SOD and CAT activity during ictus evolution, a different activity of SOD and CAT in cardioembolic and atherotrombotic infarctions, and that there is a relationship between activity of antioxidant enzymes and Mathew's score. PMID- 14762728 TI - [Role of homocysteine in the acute phase of stroke]. AB - INTRODUCTION: Hyperhomocysteinemia is a well known vascular risk factor. However its action mechanism and its role in the acute phase of stroke have not been determined. OBJECTIVES: To study plasmatic homocysteine levels in the acute stroke of different etiologies and evaluate its role as a prognostic factor. METHODS: We determined plasmatic homocysteine in 136 consecutive patients with stroke. Mean time from symptoms onset was 4.6 hours. RESULTS: Higher than normal homocysteine levels were found in ischemic stroke (n= 119 13.1 micromol/l; 43 % of patients > normality rank) and intracranial hemorrhage (n= 17 micromol/l; 24 % of patients > normality rank). The etiological distribution of ischemic stroke was the following: 42 cardioembolic (29.4%; median Hcy: 13.1 micromol/l), 20 atherothrombotic (14 %; Hcy: 12.7 micromol/l), 33 lacunar (23.1 %; Hcy: 11.8 micromol/l) and 24 undetermined (16.8%; Hcy:15.2 micromol/l). No significant differences were found between these groups (p=0.19). Those patients who presented early neurological deterioration presented higher homocysteine levels than those who remained stable or improved (median 13.3 n=16 v 11.3 n=113; p=0.061). CONCLUSIONS: High homocysteine levels are associated to all etiologic stroke subtypes and intracranial hemorrhage. Patients who experienced early neurological worsening tend to have higher homocysteine levels. PMID- 14762729 TI - [Mitochondrial disorders: a classification for the 21st century]. AB - Mitochondrial encephalomyopathies were described in the 60's when accumulation of mitochondria were observed in patients with exercise intolerance. The electronic transport chain is located in the mitochondria and forms the oxidative phosphorylation (OXPHOS) system that produces ATP in the cell. The electronic transport chain is coded by two different genomes, nuclear and mitochondrial, with different genetic characteristics. The main differences between nuclear and mitochondrial genetics are heteroplasmy and threshold effect, mitotic segregation and maternal inheritance. Mitochondrial diseases are due to defects in the genes encoding proteins involved in some mitochondrial pathways. Those genes may be ARNts, structural proteins of the electronic transport chain or associated proteins of the mitochondria (transporters or assembly proteins). Alterations in those genes may be point mutations, deletions or duplications in the mitochondrial DNA and alterations of the genomic signaling between nucleus and mitochondria. PMID- 14762730 TI - [Image of the Month. Lateral bulbar infarction with involvement of the spinal trigeminal nucleus]. PMID- 14762731 TI - [Ilioinguinal neuropathy: usefulness of conduction studies]. AB - Ilioinguinal neuropathy is an under-recognized etiology of pelvic pain, that is frequently misdiagnosed with alternative etiologies of pelvic pain. This is partially due to the limited usefulness of neurophysiological studies. Indeed, electromyography of the lower abdominal musculature identifies slightly more than half of the cases. In spite of an available conduction technique described in normal subjects, the usefulness of nerve conduction studies in ilioinguinal neuropathy is uncertain because their use has not been validated with patients. We describe the case of a patient with left inguinal pain following left inguinal herniorraphy. He underwent repeated surgeries and several analgesic treatments, without amelioration of pain. Conduction studies were consistent with ilioinguinal neuropathy. Anesthetic block relieved symptoms temporarily, and the symptoms disappeared upon section of the ilioinguinal nerve. This case describes the usefulness of conduction studies in the diagnosis of ilioinguinal neuropathy. PMID- 14762732 TI - [Buttock mass and malignant sciatic nerve tumor]. AB - Malignant peripheral nerve sheath tumors (MPNST) are rare. We report a case of a MPNST of the proximal sciatic nerve in the thigh, unassociated with pre-existing type 1 neurofibromatosis or history of radiation therapy. A 71-year-old man had a 6 month history of constant, severe, burning pain affecting the sole of the left foot. One month after the onset, the pain radiated to the left calf, posterior aspect of the thigh and buttock, and distal leg weakness followed. Three months prior to admission, the patient developed a large and painful mass in the buttock, that occupied the entire left gluteal region on examination. There was severe weakness of ankle and toe dorsiflexion and plantarflexion, decreased sensation on the lateral and posterior aspect of the left leg as well as on the dorsal and plantar surfaces of the foot, and absent ankle jerk. EMG showed denervation and motor unit loss in the short head of biceps femoris and muscles supplied by tibial and peroneal nerves on the left side. Magnetic resonance imaging revealed a 10-cm enhancing mass of the left sciatic nerve from the upper thigh to the greater sciatic notch. In surgery, a large MPNST with a high Ki67 labeling index (> 60 %) was subtotally removed from the sciatic nerve, and adjuvant radiation therapy was administered. In the ensuing months the tumor invaded the entire pelvic region. A high sciatic malignant tumor can present with a rapidly growing buttock mass and unilateral, neuropathic foot pain imitating the tarsal tunnel syndrome. PMID- 14762733 TI - [Joubert's syndrome. Presentation of two adult siblings with favorable evolution]. AB - We present the case of two siblings with Joubert's syndrome. They had breathing problems of unknown origin during the neonatal period. Their evolution has been followed-up from the first years of age until the present. They are now 26 and 22 years old, respectively. Both patients have a "borderline" mental level, are apparently normal, but have had progressive evolution and psychic and motor improvement. They can read, write and use the four operations in mathematics and can work with the computer. Both siblings are working in the familial agriculture, can play football at an elemental level with other young people of their village, and ride a bicycle and motorcycle. Both patients show normal gait and have difficulty running. Their language is normal, although somewhat slow. They perform their personal self-care that includes independent bathing, independent dressing, independent toileting and independent shaving. Their contact with other people is good. They are shy with women. They manage money. They have good social integration. One patient has bilateral strabismus and the other has unilateral ptosis. PMID- 14762734 TI - [Unemployment among neurologists. Notes for reflection and debate]. PMID- 14762735 TI - [Syphilitic cerebral arteritis: complete remission after adequate treatment]. PMID- 14762737 TI - Radioimmunotherapy with yttrium 90 ibritumomab tiuxetan (Zevalin): the role of the nuclear medicine physician. AB - Radioimmunotherapy is an emerging clinical treatment option for non-Hodgkin's lymphoma (NHL) that combines the target specificity of monoclonal antibodies with the cytotoxicity of targeted radiation, thereby avoiding the toxicity to normal organs associated with external beam radiotherapy. Yttrium 90 ibritumomab tiuxetan (Zevalin: Biogen Idec Inc, Cambridge, MA), the first radioimmunotherapeutic agent approved by the US Food and Drug Administration for the treatment of cancer, is indicated for the treatment of patients with relapsed or refractory low-grade, follicular, or transformed B-cell NHL, including patients with rituximab-refractory follicular NHL. It is likely that radioimmunotherapy will become a more common treatment option in NHL, necessitating the participation of nuclear medicine physicians on radioimmunotherapy treatment teams. The nuclear medicine physician plays a major role on the multidisciplinary treatment team, and is responsible for overseeing the administration of the radioimmunoconjugates and for interpreting the imaging scans to determine the biodistribution of the radioimmunoconjugate. It is important that nuclear medicine physicians be familiar with their role in administering radioimmunotherapy, the efficacy and safety of this treatment, and its logistics and radiation safety issues. PMID- 14762738 TI - Safety and efficacy of radioimmunotherapy with yttrium 90 ibritumomab tiuxetan (Zevalin). AB - Patients with low-grade, follicular non-Hodgkin's lymphoma usually present with advanced disease, which is not considered curable with conventional therapies. New approaches are needed to improve the outcomes in patients with this disease. Yttrium 90 ibritumomab tiuxetan (Zevalin; Biogen Idec Inc, Cambridge, MA), is highly effective, with overall response rates of 73% to 83% and complete response rates of 15% to 51%, with a median duration of response in complete responders of 23 months. The response rates tend to be higher in patients who have been treated with fewer prior therapies, and (90)Y ibritumomab tiuxetan may be suitable for use early in the course of therapy. Delayed myelosuppression is the most common adverse effect, and it is predictable, reversible, and manageable. Yttrium 90 ibritumomab tiuxetan has less nonhematologic toxicity than chemotherapy, with only minimal alopecia, mucositis, nausea, or vomiting, and a lower incidence of infections. The ibritumomab tiuxetan regimen is routinely and safely given in an outpatient setting and is completed in 7 to 9 days, and is thus more convenient for patients than chemotherapy. PMID- 14762739 TI - Imaging and dosing in radioimmunotherapy with yttrium 90 ibritumomab tiuxetan (Zevalin). AB - Yttrium 90 ibritumomab tiuxetan consists of the murine monoclonal antibody ibritumomab securely bound to the second-generation chelator tiuxetan, which attaches the high-energy pure beta emitter (90)Y, for therapy, or the gamma emitter indium 111, for imaging. The biodistribution of the therapeutic dose of (90)Y ibritumomab tiuxetan can be predicted by using an imaging dose of the antibody labeled with (111)In. Calculation of the therapeutic dose is simple and is based on patient weight and baseline platelet count: for patients with a platelet count of 150 x 10(9)/L or greater the dose of is 0.4 mCi/kg; for patients with mild thrombocytopenia (platelet count 100 x 10(9)/L to 149 x 10(9)/L) the dose is reduced to 0.3 mCi/kg; and the total dose should not exceed 32 mCi. Patients with platelet counts of less than 100 x 10(9)/L should not be treated with (90)Y ibritumomab tiuxetan. Imaging with (111)In ibritumomab tiuxetan is performed only to assess biodistribution of the radioimmunoconjugate. Uptake in sites of pathologic adenopathy, as well as other areas of lymphomatous involvement, is frequently seen on the images, but visualization of tumor uptake is not required to proceed with the therapeutic dose of (90)Y ibritumomab tiuxetan. PMID- 14762740 TI - Logistics of radioimmunotherapy with yttrium 90 ibritumomab tiuxetan (Zevalin). AB - Radioimmunotherapy is a promising new therapeutic option for the treatment of B cell non-Hodgkin's lymphoma. Several monoclonal antibody and radionuclide conjugates, including I-131 tositumomab and Y-90 ibritumomab, have been investigated in clinical trials. Yttrium 90 ibritumomab tiuxetan (Zevalin; Biogen Idec Inc, Cambridge, MA) is indicated for the treatment of patients with relapsed or refractory low-grade, follicular, or transformed B-cell non-Hodgkin's lymphoma, including patients with rituximab-refractory follicular non-Hodgkin's lymphoma. The ibritumomab tiuxetan regimen requires coordination of a multidisciplinary team on which the nuclear medicine physician (or radiation oncologist) plays a major role. The nuclear medicine physician (or radiation oncologist) is responsible for overseeing the administration of the radioimmunoconjugates and for interpreting the imaging scans to determine the expected or altered biodistribution of the radioimmunoconjugate. PMID- 14762741 TI - Radiation safety considerations with yttrium 90 ibritumomab tiuxetan (Zevalin). AB - Radioimmunotherapy is a recently approved treatment modality for non-Hodgkin's lymphoma that enables physicians to target cytotoxic localized radiation to tumor sites without using external-beam sources. Because it uses a pure beta emitter for therapy, (90)Y ibritumomab tiuxetan can be safely and routinely administered in an outpatient procedure, with few discharge instructions, and minimal risk of radiation exposure to a patient's family and acquaintances. Safety precautions for medical professionals administering (90)Y ibritumomab tiuxetan are universal precautions, with the addition of acrylic shielding for the administration of radiolabeled doses. The risk of radiation exposure to healthcare workers and family members is minimal. The primary route for biologic elimination of (90)Y ibritumomab tiuxetan is through the urine. While radiation safety instructions are not required according to the relevant patient release criteria, basic instructions to the patient and family may be valuable to further minimize the risk of radiation exposure and help alleviate patient and family concerns. PMID- 14762742 TI - New foundations in understanding osteonecrosis of the jaws. PMID- 14762743 TI - Masticatory motion after surgical or nonsurgical treatment for unilateral fractures of the mandibular condylar process. AB - PURPOSE: We sought to compare mandibular motion during mastication in patients treated in either an open or a closed fashion for unilateral fractures of the mandibular condylar process. PATIENTS AND METHODS: Eighty-one male patients with unilateral condylar process fractures were treated either with (n = 37) or without (n = 44) surgical reduction and rigid fixation of their condylar process fractures. At 6 weeks, 6 months, 1 year, and 2 years after treatment, the subjects' chewing cycles were recorded using a magnetic sensor array (Sirognathograph; Siemens Corp, Bensheim, Germany) while chewing Gummi-Bears (HARIBO, Bonn, Germany) unilaterally on the same side as the fracture and on the opposite side. The chewing cycles were analyzed using a custom computer program, and the duration, excursive ranges, and 3-dimensional cycle shape were compared between the 2 treatment groups at each time interval using multilevel linear modeling statistics. RESULTS: The 2 treatment groups did not differ significantly for any measure of cycle duration or any excursive range (except lateral excursions at 1 year post-treatment) at any of the time intervals. However, the 3 dimensional cycle shapes of the 2 groups did differ significantly at all time intervals. CONCLUSION: Surgical correction of unilateral condylar process fractures has relatively little effect on the more standard measures (duration and excursive ranges) of masticatory function. However, surgical correction better normalizes opening incisor pathways during mastication on the side opposite the fracture. PMID- 14762745 TI - Basal cell carcinoma of the maxillofacial region: site distribution and incidence rates in Arab/Jordanians, 1991 to 2000. AB - PURPOSE: We sought to estimate the incidence rates and disease trends for basal cell carcinoma (BCC) of the maxillofacial region in the civilian population in the north of Jordan. METHODS: A retrospective study was undertaken of all BCCs of the maxillofacial region diagnosed at the Regional Histopathology Department during the period 1991 to 2000. RESULTS: The age-adjusted rate for BCC in 1992 was 6.3:100,000, and in 2000, it was 8.8:100,000. However, several disease trends were observed during the study period. Age- and gender-specific incidence rates apparently increased during the study period. Lesions were most commonly seen on the nose, facial skin, and periorbital skin. The proportions of BCC in the various anatomic sites that were reviewed remained fairly constant over the 10 year period. CONCLUSIONS: In comparison with populations from other geographic areas, the incidence of BCC in the north of Jordan is low. Further studies are needed to identify behavioral and environmental factors that contribute to the development of BCC and to determine whether changes in diagnostic practices could account for the apparent rise in the incidence of this tumor in this population in recent years. PMID- 14762744 TI - Risk of osteoradionecrosis after extraction of impacted third molars in irradiated head and neck cancer patients. AB - PURPOSE: This study was performed to compare the risk of osteoradionecrosis (ORN) in head and neck cancer patients in whom 1 or more impacted third molars were extracted before radiotherapy with patients whose impacted third molars were left intact. PATIENTS AND METHODS: Eighty-one patients were selected from the medical records from 1989 to 1998. Patients had at least 1 impacted third molar and received radiotherapy for a head and neck cancer. These patients were divided into 2 groups on the basis of preirradiation extraction: group 1, patients who had impacted third molars extracted before radiotherapy (n = 55), and group 2, patients whose impacted third molars were left intact before radiotherapy (n = 38). In 12 patients of combined groups 1 and 2, at least 1 but not all of the impacted third molars were extracted before radiotherapy. RESULTS: Before radiotherapy, a total of 99 impacted third molars were extracted from the 55 patients in group 1 and a total of 55 impacted third molars were left intact in the 38 patients in group 2. After radiotherapy, a total of 7 impacted third molars were removed from 5 patients as treatment for infection (5 lower molars) or discomfort (2 upper molars). A total of 4 patients (2 from group 1 and 2 from group 2) developed ORN in the mandible. Of these 4 cases of ORN, 1 from group 1 appeared to be related to a dry socket that developed after preirradiation extraction of a lower impacted third molar, 1 from group 2 seemed to be related to infection of a lower impacted third molar after radiotherapy, and the remaining 2 cases appeared to be unrelated to an impacted third molar. CONCLUSION: Because few patients in this study developed ORN, the study failed to demonstrate whether preirradiation extraction versus retention of impacted third molars affects the risk for ORN. PMID- 14762746 TI - Ultrasound versus computed tomography in the imaging of orbital floor fractures. AB - PURPOSE: The aim of the study was to investigate whether orbital ultrasonography (US) with a curved-array transducer could be an alternative imaging method to computed tomography (CT) to detect orbital wall fractures and fractures of the infraorbital rim. MATERIALS AND METHODS: Fifty-eight patients with the clinical ophthalmologic or radiologic diagnosis of an orbital trauma were investigated prospectively by US and CT. The reference method was the intraoperative findings. RESULTS: CT evaluation of the infraorbital rim yielded a sensitivity of 79%, a specificity of 90%, and an accuracy of 94%. The positive predictive value (PPV) and the negative predictive value (NPV) of the infraorbital rim reached 69% and 83%, respectively. CT evaluation of the orbital floor showed a sensitivity of 96%, a specificity of 71%, and an accuracy of 96%. PPV and NPV resulted in 71% and 93%, respectively. US investigation of the infraorbital rim yielded a sensitivity of 77%, a specificity of 89%, and an accuracy of 97%, whereas PPV and NPV reached 65% and 83%, respectively. US investigation of the orbital floor reached a sensitivity of 94%, a specificity of 57%, and an accuracy of 96%, whereas PPV and NPV yielded 57% and 91%, respectively. No significant difference was found between US and CT in the investigation of the infraorbital rim (P =.809) and the orbital floor (P =.729). CONCLUSIONS: US with a curved-array transducer appears to be a useful alternative method in the investigation of orbital floor fractures. Further studies have to be conducted to reduce the presence of false-negative results. PMID- 14762747 TI - Mandibular condyle fractures: determinants of treatment and outcome. AB - PURPOSE: The objective of this study was to analyze the principal variables that determine the choice of the method of treatment and the outcome in condylar fractures. MATERIALS AND METHODS: We conducted a retrospective analysis of 104 mandibular condyle fractures to analyze and determine the relation between the principal clinical variables and the postoperative results. All patients underwent a clinic-radiologic investigation focusing on fracture remodeling, evolution, dental occlusion, and symmetry of the mandible. We analyzed the influence of the preoperative clinical variables (level of fracture, treatment, postoperative physical therapy, displacement and dislocation, comminution, loss of ramus height, patient age, gender, etiology, occlusion, status of dentition, and presence of facial and mandibular fractures) over the postoperative results and outcome. RESULTS: The principal factors that determined the treatment decision were the level of the fracture and the degree of displacement. The level of the fracture influenced the degree of preoperative coronal and sagittal displacement (neck fractures had greater medial and anterior displacement than head and subcondylar fractures) and the treatment applied. The functional improvement obtained by open methods was greater than that obtained by closed treatment. Open treatment increased the incidence of postoperative condylar deformities and mandibular asymmetry. CONCLUSION: The variables that influenced the method of treatment and predicted the prognosis are the level of fracture, degree and direction of displacement of the fractured segments, age, medical status of the patient, concomitant injuries, and status of dentition. PMID- 14762748 TI - Outcome assessment of patients undergoing maxillofacial procedures for the treatment of sleep apnea: comparison of subjective and objective results. AB - PURPOSE: The purpose of this study was to compare the subjective findings of the Epworth Sleepiness Scale (ESS) to the objective findings of the overnight sleep study (OSS) in 57 patients who underwent phase I and phase II surgery for the correction of obstructive sleep apnea (OSA). PATIENTS AND METHODS: Forty-two patients in phase I category (hyoid suspension, palatal surgery, and/or genioglossus advancement) and 15 patients in phase II category (maxillomandibular advancement) were examined. All patients had an OSS and completion of an ESS preoperatively and at a minimum of 8 weeks postoperatively. The results of each test were evaluated to examine any relationship between the improvements of the findings of the OSS to the changes in the ESS. RESULTS: Using accepted criteria, phase I surgery produced an 80% success rate and phase II surgery produced a greater than 95% success rate in both the respiratory disturbance index and the ESS. CONCLUSIONS: 1) Both phase I and phase II procedures are effective in treating OSA. 2) Phase II appears to be more effective in treating OSA using both objective and subjective evaluations. 3) Improvement in ESS scores and excessive daytime sleepiness seems to parallel the improvement in OSS scores in patients undergoing surgical correction of OSA. PMID- 14762749 TI - Stability of skeletal class III malocclusion after combined maxillary and mandibular procedures: rigid internal fixation versus wire osteosynthesis of the mandible. AB - PURPOSE: The aim of this study was to evaluate skeletal stability after double jaw surgery for correction of skeletal Class III malocclusion to assess whether there were any differences between wire and rigid fixation of the mandible. PATIENTS AND METHODS: Thirty-seven Class III patients had Le Fort I osteotomy stabilized with plate and screws for maxillary advancement. Bilateral sagittal split osteotomy for mandibular setback was stabilized with wire osteosynthesis and maxillomandibular fixation for 6 weeks in 20 patients (group 1) and with rigid internal fixation in 17 patients (group 2). Lateral cephalograms were taken before surgery, immediately after surgery, 8 weeks after surgery, and 1 year after surgery. RESULTS: Before surgery, both groups were balanced with respect to linear and angular measurements of craniofacial morphology. One year after surgery, maxillary sagittal stability was excellent in both groups, and bilateral sagittal split osteotomy accounted for most of the total horizontal relapse observed. In group 1, significant correlations were found between maxillary advancement and relapse at the posterior maxilla and between mandibular setback and postoperative counterclockwise rotation of the ramus and mandibular relapse. In group 2, significant correlations were found between mandibular setback and intraoperative clockwise rotation of the ramus and between mandibular setback and postoperative counterclockwise rotation of the ramus and mandibular relapse. No significant differences in postoperative skeletal and dental stability between groups were observed except for maxillary posterior vertical position. CONCLUSIONS: Surgical correction of Class III malocclusion after combined maxillary and mandibular procedures appears to be a fairly stable procedure independent of the type of fixation used to stabilize the mandible. PMID- 14762750 TI - Clinical outcome of third molars in adults followed during 18 years. AB - PURPOSE: The aim of the present study was to follow the clinical changes in third molar status during an 18-year period in patients aged 20 to 38 years. PATIENTS AND METHODS: The series consisted of 118 subjects (37 men and 81 women). In the beginning of the study, the mean age was 20.2 years (SD, +/-0.6 year), and at the end, it was 38.6 years (SD, +/-0.6 year). Panoramic radiographs were taken at baseline and at age 38. All of the subjects were clinically examined at baseline and at the end of the study. A portion of the subjects (n = 69) were also examined at age 32. RESULTS: Most of the initially unerupted third molars were removed during the follow-up period (73%, maxilla and mandible together). More than half of the initially partially erupted third molars were removed during the follow-up period (64%, maxilla and mandible together). The percentage of erupted third molars found in the mouth at age 38 increased significantly depending on the initial status. Of the initially unerupted, partially erupted, or erupted third molars, 10%, 33%, and 50%, respectively, were erupted at age 38 (maxilla and mandible together). Changes in the status of third molars continued from age 32 to age 38, although to a lesser extent (8%). The 3 third molars with advanced eruption were all maxillary teeth in men. CONCLUSION: Third molars undergo continuous clinical change on a reduced scale at least up to the age of 38 years. PMID- 14762751 TI - Comparative study of locking plates in mandibular reconstruction after ablative tumor surgery: THORP versus UniLOCK system. AB - PURPOSE: This retrospective study compared 2 types of rigid locking plates (THORP and UniLOCK; Stratec Medical, Oberdorf, Switzerland) in 107 patients undergoing reconstruction for bridging mandibular defects following ablative tumor surgery. PATIENTS AND METHODS: From January 1993 to December 2000, 107 patients were reconstructed following ablative tumor surgery with 57 THORP plates and 50 UniLOCK plates. Study follow-up ranged from 18 to 87 months (average, 32 months). Complications were categorized into delayed wound healing, infection, plate exposure, and plate fracture, taking into account the type and timing of reconstruction. RESULTS: Overall type and number of complications show no statistically significant differences between THORP and UniLOCK groups. Infection was the most frequent type of complication (THORP, 30; UniLOCK, 32). Others included delayed wound healing (THORP, 13; UniLOCK, 12), plate exposure (THORP, 8; UniLOCK, 7), and plate fracture (THORP, 5; UniLOCK, 1). Plates were removed in 22 THORP and 11 UniLOCK plates. The most frequent reason for plate removal in both groups was infection. Other reasons for plate removal include tumor recurrence, plate fracture, plate exposure, or a combination of reasons. CONCLUSIONS: THORP and UniLOCK plates do not present statistically significant differences in the parameters studied. Nevertheless, the UniLOCK group had slightly better results. Considering that the THORP system is much bulkier and its screws bigger, our results lead to the conclusion that bridging osteosynthesis with a 2.4 UniLOCK system is adequate for plate reconstruction of mandibular defects. PMID- 14762752 TI - Ultrasound to stimulate mandibular bone defect healing: a placebo-controlled single-blind study in rats. AB - PURPOSE: Because of the limitations of the body to heal large maxillofacial bone defects, an attempt was made to stimulate mandibular defect healing with low intensity pulsed ultrasound in rats. This ultrasound consists of a 1.5-MHz pressure wave administered in pulses of 200 microsec, with an average intensity over space and time of 30 mW. cm(-2). MATERIALS AND METHODS: In 72 rats, a 5.0-mm diameter circular mandibular defect was created. Three groups were studied: an ultrasound treatment group, a placebo treatment group, and a control group. Ultrasound and placebo treatment involved a daily treatment for 20 minutes at the site of the defect under general anesthesia. At 2 and 4 weeks, the region of bone growth within the defect was measured using microradiographs and the amount of defect healing was expressed as the percentage of defect closure. RESULTS: At 2 and 4 weeks, there was no statistical significant difference in the percentage of defect closure between the groups. CONCLUSION: Low-intensity pulsed ultrasound does not stimulate bone defect healing in the case of a large mandibular defect in the rat. PMID- 14762753 TI - Histomorphometric evaluation of bone regeneration using allogeneic and alloplastic bone substitutes. AB - PURPOSE: The purpose of this investigation was to assess bone regeneration in critical sized defects in the rabbit calvarium using allogeneic and alloplastic bone substitutes. MATERIALS AND METHODS: Thirty New Zealand White rabbits were divided into 3 groups of 10 animals each. Bilateral 15 mm x 17 mm calvarial defects were made in the parietal bones of each animal. Group 1 had demineralized bone matrix (DBM) gel placed in one defect, while the other defect was left unfilled and served as the control. Group 2 had one defect filled with calcium hydroxide (CaOH)-treated DBM gel and the other defect filled with DBM gel. Group 3, the calcium-phosphate cement group, had Norian CRS (Norian Corp, Cupertino, CA) placed on one side and Bone Source (Howmedica Leibinger, Dallas, TX) placed on the contralateral side. Five animals in each group were killed at 6 and 12 weeks. Data analysis included qualitative assessment of the calvarial specimens and radiographic evaluation. Histomorphometric analysis was used to quantify the amount of new bone within the defects. RESULTS: Histomorphometric analysis showed that DBM gel-treated defects had significantly more new bone at 12 weeks compared with all other groups. There was no significant difference between defects filled with CaOH-treated DBM gel and those filled with DBM gel at 12 weeks. In group 3, Norian CRS- and Bone Source-treated defects were not statistically different from the unfilled controls. CONCLUSION: DBM gel was an effective allogeneic bone substitute that showed reliable osseous healing of critical size defects in the rabbit calvarium. The addition of CaOH to DBM gel did not significantly improve the bone regenerative capacity of the DBM gel. Both Norian CRS and Bone Source did not promote bone regeneration in this animal model. PMID- 14762755 TI - Clinical applications of fibrin sealants. PMID- 14762754 TI - Rapidly growing mass in mandibular gingiva. PMID- 14762756 TI - Massive osteolysis (Gorham disease) of the maxillofacial skeleton: report of 2 cases. PMID- 14762757 TI - Invasive fungal infection of the midfacial and orbital complex due to Scedosporium apiospermum and mucormycosis. PMID- 14762758 TI - Injectable calcium-phosphate bone cement (Norian) for reconstruction of a large mandibular defect: a case report. PMID- 14762760 TI - Recurrent temporomandibular joint dislocation treated with botulinum toxin: report of 3 cases. PMID- 14762759 TI - Acute hypertensive crisis resulting in pulmonary edema and myocardial ischemia during orthognathic surgery. PMID- 14762761 TI - Head and neck injury from a leopard attack: case report and review of the literature. PMID- 14762762 TI - Degradation rate of delta (resorbable) internal fixation: report of 2 cases. PMID- 14762763 TI - Pyriform rim sandwich osteotomy: a new regional osteotomy for correction of para alar deficiency. PMID- 14762764 TI - Comparison of 2 temporomandibular joint total joint prosthesis systems. PMID- 14762765 TI - Comparison of 2 temporomandibular joint total joint prosthesis systems. PMID- 14762771 TI - Mixed potential impact on endoscopy centers of Medicare bill. PMID- 14762773 TI - The NIH roadmap for medical research: bypassing detours. PMID- 14762774 TI - Image of the month. Diverticulitis of the sigmoid colon. PMID- 14762775 TI - Use of 5-fluorouracil and survival in patients with microsatellite-unstable colorectal cancer. AB - BACKGROUND & AIMS: 5-Fluorouracil improves mortality in stage III colorectal cancer patients. In vitro studies suggest that microsatellite instability influences cell survival after 5-fluorouracil treatment. We investigated the survival influence of 5-fluorouracil in patients with microsatellite instability high tumors. METHODS: We collected data and tumors on 204 consecutive stage II and III colorectal cancer patients from registries at the University of California and Veterans Administration hospitals in San Diego, California, from 1982 to 1999. Archival DNA was extracted, and microsatellite instability was assessed by National Cancer Institute-recommended markers. Cox proportional hazard modeling was used to determine survival associations for microsatellite instability and 5-fluorouracil treatment status. RESULTS: We identified 36 microsatellite instability-high (17.6%) and 168 non-microsatellite instability high tumors (82.4%). Microsatellite instability-high tumors were significantly associated with proximal colon location, presence of mucin, and surrounding lymphoid reaction. Univariate and multivariate analyses showed no survival difference between microsatellite instability-high and non-microsatellite instability-high groups (hazard ratio, 1.04; P = 0.88). Dichotomized by use of 5 fluorouracil, there was increased risk of death in patients who received no adjuvant chemotherapy (hazard ratio, 2.02; P = 0.02). However, the benefit of 5 fluorouracil was different between microsatellite instability-high and non microsatellite instability-high groups. Patients with non-microsatellite instability-high tumors who received 5-fluorouracil had better survival compared with patients who were not treated (P < 0.05). Conversely, patients with microsatellite instability-high tumors who were treated with 5-fluorouracil had no survival difference compared with patients without treatment (P = 0.52). CONCLUSIONS: There is improved survival in patients with non-microsatellite instability-high tumors after 5-fluorouracil-based chemotherapy that does not extend to patients with microsatellite instability-high tumors. The microsatellite instability status of a patient's colorectal cancer may indicate differences in 5-fluorouracil-based chemosensitivity; this is consistent with in vitro studies. PMID- 14762776 TI - Comparison of scheduled and episodic treatment strategies of infliximab in Crohn's disease. AB - BACKGROUND & AIMS: This analysis of Crohn's disease patients treated with infliximab in ACCENT I compared episodic and scheduled treatment strategies under conditions that simulate clinical practice. METHODS: After 5 mg/kg infliximab at week 0, 573 patients were randomized to infusions at weeks 2 and 6 and every 8 weeks until week 46 of placebo (episodic), infliximab 5 mg/kg at weeks 2 and 6 followed by 5 mg/kg (5 mg/kg scheduled) every 8 weeks, or infliximab 5 mg/kg at weeks 2 and 6 followed by 10 mg/kg (10 mg/kg scheduled) every 8 weeks. At or after week 14, treatment could be given with a dose of infliximab 5 mg/kg higher upon loss of response. RESULTS: The efficacy of scheduled infliximab therapy was better than episodic treatment. Crohn's Disease Activity Index (CDAI) scores were consistently significantly better in the 10 mg/kg scheduled maintenance group from weeks 10 to 54, and response and remission rates (combined scheduled) were significantly higher from weeks 10 to 30. A greater proportion of patients achieved complete mucosal healing at week 54 (P = 0.041). A lower proportion developed antibodies to infliximab in the scheduled groups than in the episodic group (9% [5 mg/kg], 6% [10 mg/kg], 28% [episodic], respectively). Scheduled strategy patients had fewer Crohn's disease-related hospitalizations (P = 0.014) and surgeries (P = 0.01) than episodic strategy patients. CONCLUSIONS: The scheduled infliximab groups, particularly the 10 mg/kg group, had better CDAI and Inflammatory Bowel Disease Questionnaire (IBDQ) responses than those in the episodic group. Both scheduled groups had fewer hospitalizations, higher rates of mucosal healing, and fewer developed antibodies than those in the episodic group, with no increase in side effects. PMID- 14762777 TI - Association of antibody responses to microbial antigens and complications of small bowel Crohn's disease. AB - BACKGROUND & AIMS: Crohn's disease patients can be characterized by antibody responses against Crohn's disease-related bacterial sequence, Escherichia coli outer membrane porin C, Saccharomyces cerevisiae (oligomannan), and neutrophil nuclear antigens. Our aim was to determine whether expression of antibodies against Crohn's disease-related bacterial sequence and Escherichia coli outer membrane porin C is associated with distinct phenotypic manifestations. METHODS: Sera from 303 patients were tested for antibodies to the Crohn's disease-related bacterial sequence (I2), anti-Escherichia coli outer membrane porin C, anti Saccharomyces cerevisiae, and perinuclear antineutrophil cytoplasmic antibodies and for 3 Crohn's disease-associated variants of the NOD2 gene (R702W, G908R, and 1007fs) and compared with clinical data. RESULTS: Patients expressing I2 were more likely to have fibrostenosing Crohn's disease (64.4% vs. 40.7%; P < 0.001) and to require small bowel surgery (62.2% vs. 37.4%; P < 0.001). Patients with anti-Escherichia coli outer membrane porin C were more likely to have internal perforating disease (50.0% vs. 30.7%; P = 0.001) and to require small bowel surgery (61.4% vs. 44.2%; P = 0.003). Anti-Crohn's disease-related bacterial sequence was independently associated with fibrostenosis (P = 0.027) and small bowel surgery (P = 0.01), whereas anti-Escherichia coli outer membrane porin C was independently associated with internal perforations (P < 0.006). Patients positive for I2, anti-Escherichia coli outer membrane porin C, and anti Saccharomyces cerevisiae were the most likely to have undergone small bowel surgery (72.0%; odds ratio, 8.6; P < 0.001) compared with patients without reactivity (23.0%). When the presence and magnitude of antibody responses were considered, 90% of patients with small bowel disease who required surgery had high levels of I2, Escherichia coli outer membrane porin C, and oligomannan antibodies, compared with only 18.2% with low-titer responses (P < 0.001). CONCLUSIONS: I2 and anti-Escherichia coli outer membrane porin C are associated with Crohn's disease phenotypes, and patients with the highest level of serum reactivity toward an increasing number of microbiota have the greatest frequency of strictures, internal perforations, and small bowel surgery. PMID- 14762780 TI - Independent influences of body mass and gastric volumes on satiation in humans. AB - BACKGROUND & AIMS: We assessed the association of body mass and gastric volumes (fasting and postprandial) with satiation and postprandial symptoms. METHODS: Healthy obese and nonobese subjects underwent measurement of caloric intake at maximum satiation; postprandial symptoms were measured with visual analogue scales 30 minutes after a meal. Gastric volume during fasting and after 300 mL of Ensure was measured with technetium-99m single-photon emission computed tomography imaging. We used multiple regression analysis to assess the associations among variables. RESULTS: Among 134 participants (81 women and 53 men), the median age was 26 years (range, 12-58 years), and the median body mass index was 24 kg/m(2) (range, 17-48 kg/m(2)). Increased body mass index, but not height, was associated with delayed satiation (P < 0.003, adjusted for sex). Overweight and obese subjects ingested, on average, 225 +/- 57 more kilocalories (945 +/- 239 kJ) at maximum satiation compared with normal weight individuals. Increased fasting gastric volume was not associated with body mass index or height, but it was significantly associated with delayed satiation (P = 0.001, adjusted for body mass index and sex). An increase of 50 mL in the fasting gastric volume was associated with 114 +/- 32 kcal (479 +/- 134 kJ) more ingested at maximum satiation. Increased body mass index was associated with lower fullness scores 30 minutes after a meal (P = 0.0012, adjusted for sex and volume of Ensure ingested). In contrast, scores of postprandial bloating and pain were higher with increased body mass index (both P < 0.05, adjusted for sex and volume of Ensure ingested). CONCLUSIONS: Greater body mass index and fasting gastric volume are associated with reduced satiation. Increased body mass index or height was not associated with greater gastric volumes. PMID- 14762779 TI - Prostanoids, ornithine decarboxylase, and polyamines in primary chemoprevention of familial adenomatous polyposis. AB - BACKGROUND & AIMS: Familial adenomatous polyposis because of germline mutation of the adenomatous polyposis coli gene is characterized by development of colorectal adenomas and, ultimately, colorectal cancer. The usefulness of colorectal mucosal compounds to predict the effect on adenoma development of primary chemoprevention with the nonsteroidal anti-inflammatory drug sulindac was evaluated. METHODS: A randomized, double-blind, placebo-controlled study of 41 subjects genotypically affected with familial adenomatous polyposis but phenotypically unaffected was conducted. Patients received either sulindac or placebo for 48 months, and development of new adenomas was evaluated. The levels of 5 prostanoids, ornithine decarboxylase, and polyamines were measured serially in normal-appearing rectal mucosa. RESULTS: There were no statistically significant differences between treatment groups in baseline levels of prostanoids, ornithine decarboxylase, or polyamines. At conclusion of the study, 4 of 5 prostaglandin levels were statistically significantly lower in the sulindac group than in the placebo group. Among the subset of patients taking sulindac, 3 of 5 prostaglandin levels were statistically significantly lower in patients who were polyp free than in those who developed polyps. By contrast, there were no statistically significant differences in ornithine decarboxylase or polyamines between treatment groups or in those on sulindac who were polyp free compared with those who developed polyps. CONCLUSIONS: Colorectal mucosal prostaglandin levels, but not ornithine decarboxylase or polyamines, may be valuable biomarkers to assess appropriate drug dosage and medication compliance in patients undergoing primary chemoprevention therapy with sulindac. Reduction of mucosal prostaglandin levels may be necessary to achieve chemopreventive benefit from this agent. PMID- 14762781 TI - Addition of a second endoscopic treatment following epinephrine injection improves outcome in high-risk bleeding ulcers. AB - BACKGROUND & AIMS: Endoscopic therapy reduces the rebleeding rate, the need for surgery, and the mortality in patients with peptic ulcer and active bleeding or visible vessel. Injection of epinephrine is the most popular therapeutic method. Guidelines disagree on the need for a second hemostatic procedure immediately after epinephrine; although it seems to reduce further bleeding, its effects on morbidity, surgery rates, and mortality remain unclear. The aim of this study was to perform a systematic review and meta-analysis to determine whether the addition of a second procedure improves hemostatic efficacy and/or patient outcomes after epinephrine injection. METHODS: An extensive search for randomized trials comparing epinephrine alone vs. epinephrine plus a second method was performed in MEDLINE and EMBASE and in the abstracts of the AGA Congresses between 1990 and 2002. Selected articles were included in a meta-analysis. RESULTS: Sixteen studies including 1673 patients met inclusion criteria. Adding a second procedure reduced the further bleeding rate from 18.4% to 10.6% (Peto odds ratio 0.53, 95% CI: 0.40-0.69) and emergency surgery from 11.3% to 7.6% (OR: 0.64, 95% CI: 0.46-0.90). Mortality fell from 5.1% to 2.6% (OR: 0.51, 95% CI: 0.31-0.84). Subanalysis showed that the risk of further bleeding decreased regardless of which second procedure was applied. In addition, the risk was reduced in all subgroups, although reduction was more evident in high-risk patients and when no scheduled follow-up endoscopies were performed. CONCLUSIONS: Additional endoscopic treatment after epinephrine injection reduces further bleeding, need for surgery, and mortality in patients with bleeding peptic ulcer. PMID- 14762782 TI - Severity of inflammation is a risk factor for colorectal neoplasia in ulcerative colitis. AB - BACKGROUND & AIMS: Patients with ulcerative colitis are at increased risk of colorectal cancer. It is widely believed that this is secondary to colonic inflammation. However, the severity of colonic inflammation has never been shown to be a risk factor. METHODS: We devised a case-control study of patients with long-standing extensive ulcerative colitis to examine various potential risk factors for neoplasia. All cases of colorectal neoplasia detected from our surveillance program between January 1, 1988, and January 1, 2002, were studied (n = 68). Each patient was matched with 2 control patients from the same surveillance population (n = 136). Matching was for sex, colitis extent, age at onset, duration of colitis, and year of index surveillance colonoscopy. Segmental colonoscopic and histological inflammation was recorded by using a simple score (0, normal; 1, quiescent/chronic inflammation; and 2, 3, and 4, mild, moderate, and severe active inflammation, respectively). Other data collected included history of primary sclerosing cholangitis, family history of colorectal cancer, and smoking and drug history (mesalamine 5-aminosalicylic acid, azathioprine, and folate). RESULTS: Univariate analysis showed a highly significant correlation between the colonoscopic (odds ratio, 2.5; P = 0.001) and histological (odds ratio, 5.1; P < 0.001) inflammation scores and the risk of colorectal neoplasia. No other factors reached statistical significance. On multivariate analysis, only the histological inflammation score remained significant (odds ratio, 4.7; P < 0.001). CONCLUSIONS: In long-standing extensive ulcerative colitis, the severity of colonic inflammation is an important determinant of the risk of colorectal neoplasia. Endoscopic and histological grading of inflammation could allow better risk stratification for surveillance programs. PMID- 14762783 TI - Diabetes increases the risk of chronic liver disease and hepatocellular carcinoma. AB - BACKGROUND & AIMS: An association between diabetes and chronic liver disease has been reported. However, the temporal relationship between these conditions remains unknown. METHODS: We identified all patients with a hospital discharge diagnosis of diabetes between 1985 and 1990 using the computerized records of the Department of Veterans Affairs. We randomly assigned 3 patients without diabetes for every patient with diabetes. We excluded patients with concomitant liver disease. The remaining cohort was followed through 2000 for the occurrence of chronic nonalcoholic liver disease (CNLD) and hepatocellular carcinoma (HCC). Hazard rate ratios (HRR) were determined in Cox proportional hazard survival analysis. RESULTS: The study cohort comprised 173,643 patients with diabetes and 650,620 patients without diabetes. Most were men (98%). Patients with diabetes were older (62 vs. 54 years) than patients without diabetes. The incidence of chronic nonalcoholic liver disease was significantly higher among patients with diabetes (incidence rate: 18.13 vs. 9.55 per 10,000 person-years, respectively, P < 0.0001). Similar results were obtained for HCC (incidence rate: 2.39 vs. 0.87 per 10,000 person-years, respectively, P < 0.0001). Diabetes was associated with an HRR of 1.98 (95% CI: 1.88 to 2.09, P < 0.0001) of CNLD and an HRR of 2.16 (1.86 to 2.52, P < 0.0001) of hepatocellular carcinoma. Diabetes carried the highest risk among patients with longer than 10 years of follow-up. CONCLUSIONS: Among men with diabetes, the risk of CNLD and HCC is doubled. This increase in risk is independent of alcoholic liver disease, viral hepatitis, or demographic features. PMID- 14762784 TI - Improved clinical outcome using polytetrafluoroethylene-coated stents for TIPS: results of a randomized study. AB - BACKGROUND & AIMS: A 50% dysfunction rate at 1 year is one of the main drawbacks of the transjugular intrahepatic portosystemic shunt procedure. Preliminary experimental and clinical studies suggest that the use of stents covered with polytetrafluoroethylene could tremendously decrease this risk. METHODS: Eighty patients with cirrhosis and uncontrolled bleeding (n = 23), recurrent bleeding (n = 25), or refractory ascites (n = 32) were randomized to be treated by transjugular intrahepatic portosystemic shunts with either a polytetrafluoroethylene-covered stent (group 1; 39 patients) or a usual uncovered prosthesis (group 2; 41 patients). Follow-up Doppler ultrasound was scheduled at day 7, at 1 month, and then every 3 months for 2 years. Angiography and portosystemic pressure gradient measurements were performed 6, 12, and 24 months after the transjugular intrahepatic portosystemic shunt procedure and whenever dysfunction was suspected. Dysfunction was defined as a >50% reduction of the lumen of the shunt at angiography or a portosystemic pressure gradient >12 mm Hg. RESULTS: After a median follow-up of 300 days, 5 patients (13%) in group 1 and 18 (44%) in group 2 experienced shunt dysfunction (P < 0.001). Clinical relapse occurred in 3 patients (8%) in group 1 and 12 (29%) in group 2 (P < 0.05). Actuarial rates of encephalopathy were 21% in group 1 and 41% in group 2 at 1 year (not significant). Estimated probabilities of survival were 71% and 60% at 1 year and 65% and 41% at 2 years in groups 1 and 2, respectively (not significant). CONCLUSIONS: The use of polytetrafluoroethylene-covered prostheses improves transjugular intrahepatic portosystemic shunt patency and decreases the number of clinical relapses and reinterventions without increasing the risk of encephalopathy. PMID- 14762785 TI - Altered control of gastric acid secretion in gastrin-cholecystokinin double mutant mice. AB - BACKGROUND & AIMS: Three pathways control gastric acid secretion: the gastrin enterochromaffin-like (ECL) cell axis, the vagus-parietal cell axis, and the cholecystokinin (CCK)-D cell axis. Mice lacking gastrin or both gastrin and CCK were examined to determine the role of the hormones. METHODS: Acid was measured after pylorus ligation, and biopsies from gastrin knockout (KO), gastrin-CCK double-KO, and wild-type (WT) mice were collected for biochemical, immunocytochemical, and electron-microscopic examination. RESULTS: The ECL cells were inactive in both groups of mutant mice but the cell number was unaffected. Both parietal cell number and level of H(+)/K(+)-ATPase messenger RNA (mRNA) were reduced in the mutant strains, but gastrin-CCK double-KO mice displayed more active parietal cells and larger acid output than the gastrin KO mice. The acid response to histamine in double-KO mice was unchanged whereas that to gastrin was diminished, but it could be restored by infusion of gastrin. Oxyntic D-cell density was the same in both mutant strains, but the D cells were more active in the gastrin KO than in the double-KO mice. CCK infusion in gastrin-CCK double-KO mice raised the somatostatin mRNA level and inhibited acid secretion to the level seen in gastrin KO mice. Vagotomy and atropine abolished acid secretion in all 3 groups of mice. CONCLUSIONS: Lack of gastrin impairs the gastrin-ECL axis, whereas lack of gastrin and CCK impairs both hormonal pathways. In the gastrin CCK double-KO mice, acid secretion is only controlled by cholinergic vagal stimulation, which normalizes the acid output. PMID- 14762786 TI - High-molecular-weight polyethylene glycol prevents lethal sepsis due to intestinal Pseudomonas aeruginosa. AB - BACKGROUND & AIMS: During stress, erosion of protective intestinal mucus occurs in association with adherence to and disruption of the intestinal epithelial barrier by invading opportunistic microbial pathogens. The aims of this study were to test the ability of a high-molecular-weight polyethylene glycol compound, polyethylene glycol 15-20, to protect the intestinal epithelium against microbial invasion during stress. METHODS: The ability of polyethylene glycol 15-20 to protect the intestinal epithelium against the opportunistic pathogen Pseudomonas aeruginosa was tested in cultured Caco-2 cells. Bacterial virulence gene expression, bacterial adherence, and transepithelial electrical resistance were examined in response to apical inoculation of P. aeruginosa onto Caco-2 cells. Complementary in vivo studies were performed in a murine model of lethal sepsis due to intestinal P. aeruginosa in which surgical stress (30% hepatectomy) was combined with direct inoculation of P. aeruginosa into the cecum. RESULTS: High molecular-weight polyethylene glycol (polyethylene glycol 15-20) conferred complete protection against the barrier-dysregulating effects of P. aeruginosa in Caco-2 cells. Intestinal application of polyethylene glycol 15-20 in stressed mice protected against the lethal effects of intestinal P. aeruginosa. Mechanisms of this effect seem to involve the ability of polyethylene glycol 15-20 to distance P. aeruginosa from the intestinal epithelium and render it completely insensate to key environmental stimuli that activate its virulence. CONCLUSIONS: High-molecular-weight polyethylene glycol has the potential to function as a surrogate mucin within the intestinal tract of a stressed host by inhibiting key interactive events between colonizing microbes and their epithelial cell targets. PMID- 14762787 TI - Leptin reduces the development of the initial precancerous lesions induced by azoxymethane in the rat colonic mucosa. AB - BACKGROUND & AIMS: Recent studies suggest that leptin, a hormone involved in food intake regulation, released into the circulation and gastrointestinal juice, may be a growth factor for intestine and may be involved in carcinogenesis; however, data are contradictory. This study investigates in rat colonic mucosa (1) the effects of hyperleptinemia on epithelial cell proliferation and development of aberrant crypts, earliest preneoplastic lesions, and (2) whether luminal leptin affects cell proliferation. METHODS: Leptin (1 mg/kg/d) or vehicle was administered systemically by miniosmotic pump in Fischer 344 rats either for 7 days (BrdU-labeling indices study) or 23 days (azoxymethane-induced colonic lesions study). The effects of injections or continuous infusion of leptin into the colon were also studied. RESULTS: In systemic leptin-treated rats, plasma leptin levels were 4- to 5-fold increased (P < 0.008 to P < 0.001); labeling indices were higher in proximal colon than in pair-fed control rats (P = 0.006) but unaffected in distal colon. Unexpectedly, in azoxymethane-treated rats, leptin significantly inhibited aberrant crypt foci formation in the middle and distal colon compared with controls (P = 0.006). Under these conditions, plasma insulin levels were reduced by 41%-58%, but gastrin levels were unchanged. In controls, luminal immunoreactive leptin reached the colon. A 3.6-fold increase in intraluminal leptin had no effect on epithelial cell proliferation. CONCLUSIONS: This study provides the first evidence that leptin reduces the development of chemically induced precancerous lesions in colon, perhaps through decreased insulinemia, and thus does not support an important role for leptin in carcinogenesis promotion. Moreover, the study indicates that leptin is not a potent growth factor for normal intestine. PMID- 14762788 TI - Prevention of toxin-induced intestinal ion and fluid secretion by a small molecule CFTR inhibitor. AB - BACKGROUND & AIMS: The cystic fibrosis transmembrane conductance regulator (CFTR) provides an important apical route for Cl(-) secretion across intestinal epithelia. A thiazolidinone-type CFTR blocker (CFTR(inh)-172) reduced cholera toxin-induced fluid accumulation in mouse intestinal loops. Here, we characterize the efficacy and pharmacodynamics of CFTR(inh)-172 in blocking cAMP and cGMP induced Cl(-)/fluid secretion in rodent and human intestine. METHODS & RESULTS: CFTR(inh)-172 inhibited cAMP and cGMP agonist induced short-circuit current by >95% in T84 colonic epithelial cells (K(I) approximately 3 micromol/L) and in mouse and human intestinal sheets (K(I) approximately 9 micromol/L). A single intraperitoneal injection of CFTR(inh)-172 (200 microg) blocked intestinal fluid secretion in a rat closed-loop model by >90% for cholera toxin and >70% for STa Escherichia coli toxin. In mice, CFTR(inh)-172 (20 microg) inhibited cholera toxin-induced intestinal fluid secretion by 90% (persistence t(1/2) approximately 10 hours, K(I) approximately 5 microg) and STa toxin by 75% (K(I) approximately 10 microg). Tissue distribution and pharmacokinetic studies indicated intestinal CFTR(inh)-172 accumulation facilitated by enterohepatic circulation. An oral CFTR(inh)-172 preparation reduced fluid secretion by >90% in a mouse open-loop cholera model. CONCLUSIONS: A small molecule CFTR blocker markedly reduced intestinal ion and fluid secretion caused by cAMP/cGMP-dependent bacterial enterotoxins. CFTR inhibition may thus reduce fluid secretion in infectious secretory diarrheas. PMID- 14762789 TI - Toll-like receptor 9 signaling mediates the anti-inflammatory effects of probiotics in murine experimental colitis. AB - BACKGROUND & AIMS: We tested whether the attenuation of experimental colitis by live probiotic bacteria is due to their immunostimulatory DNA, whether toll-like receptor (TLR) signaling is required, and whether nonviable probiotics are effective. METHODS: Methylated and unmethylated genomic DNA isolated from probiotics (VSL-3), DNAse-treated probiotics and Escherichia coli (DH5 alpha) genomic DNA were administered intragastrically (i.g.) or subcutaneously (s.c.) to mice prior to the induction of colitis. Viable or gamma-irradiated probiotics were administered i.g. to wild-type mice and mice deficient in different TLR or in the adaptor protein MyD88, 10 days prior to administration of dextran sodium sulfate (DSS) to their drinking water and for 7 days thereafter. RESULTS: Intragastric and s.c. administration of probiotic and E. coli DNA ameliorated the severity of DSS-induced colitis, whereas methylated probiotic DNA, calf thymus DNA, and DNase-treated probiotics had no effect. The colitis severity was attenuated to the same extent by i.g. delivery of nonviable gamma-irradiated or viable probiotics. Mice deficient in MyD88 did not respond to gamma-irradiated probiotics. The severity of DSS-induced colitis in TLR2 and TLR4 deficient mice was significantly decreased by i.g. administration of gamma-irradiated probiotics, whereas, in TLR9-deficient mice, gamma-irradiated probiotics had no effect. CONCLUSIONS: The protective effects of probiotics are mediated by their own DNA rather than by their metabolites or ability to colonize the colon. TLR9 signaling is essential in mediating the anti-inflammatory effect of probiotics, and live microorganisms are not required to attenuate experimental colitis because nonviable probiotics are equally effective. PMID- 14762790 TI - Hepatitis C virus core and nonstructural proteins induce fibrogenic effects in hepatic stellate cells. AB - BACKGROUND & AIMS: The mechanisms by which hepatitis C virus (HCV) induces liver fibrosis are unknown. Hepatocytes secrete HCV proteins, which may interact with hepatic stellate cells (HSCs). Our aims were to investigate whether HCV proteins induce fibrogenic effects on HSCs. METHODS & RESULTS: Human-activated HSCs expressed messenger RNA (mRNA) for the putative HCV receptors CD81, LDL receptor, and C1q receptor as assessed by RT-PCR. Incubation of activated but not quiescent human HSCs with recombinant core and NS3 protein increased intracellular calcium concentration and reactive oxygen species production, as well as stimulated intracellular signaling pathways. Adenoviruses encoding core and nonstructural proteins (NS3-NS5) were used to express HCV proteins in HSCs. Expression of core protein increased cell proliferation in a Ras/ERK and PI3K/AKT dependent manner. In contrast, NS3-NS5 protein expression preferentially induced proinflammatory actions, such as increased chemokine secretion and expression of intercellular cell adhesion molecule type 1 (ICAM-1) through the NF-kappa B and c-Jun N terminal kinase pathways. These effects were attenuated by antioxidants. Infection of freshly isolated rat HSCs with adenovirus-encoding core protein resulted in accelerated cell activation, as assessed by alpha-smooth muscle actin expression. Moreover, adenovirus-encoding core and NS3-NS5 proteins increased the secretion of bioactive TGF beta 1 and the expression of procollagen alpha1(I) in early cultured rat HSCs, as assessed by ELISA and RNase protection assay, respectively. CONCLUSIONS: HCV core and nonstructural proteins regulate distinct biologic functions in HSCs. A direct interaction between HCV proteins and HSCs may contribute to HCV-induced liver fibrosis. PMID- 14762791 TI - Trafficking of the bile salt export pump from the Golgi to the canalicular membrane is regulated by the p38 MAP kinase. AB - BACKGROUND & AIMS: Bile secretion depends on the delivery and removal of transporter proteins to and from the canalicular membrane. Trafficking of the bile salt export pump (BSEP) to the canalicular membrane was investigated in HepG2 cells and rat hepatocytes. METHODS: Subcellular localization of BSEP was determined by confocal laser scanning microscopy using different BSEP antibodies. RESULTS: Ten percent of untreated HepG2 cells developed pseudocanaliculi, but only 15% of these pseudocanaliculi contained BSEP, which largely colocalized with the Golgi marker GM130. Cycloheximide, an inhibitor of protein translation, induced a microtubule- and p38(MAP) kinase-dependent decrease of Golgi-associated BSEP, accompanied by a more than 2-fold increase in BSEP-positive pseudocanaliculi. Also, tauroursodeoxycholate (TUDC), which activates p38(MAP) kinase (p38(MAPK), increased BSEP-positive pseudocanaliculi by more than 50% in rat sodium taurocholate cotransporting peptide (Ntcp)-transfected but not in untransfected HepG2 cells. The TUDC-dependent increase was sensitive to inhibitors of p38(MAPK) and microtubules and involved Ca(2+)-independent protein kinase C isoforms as suggested by its sensitivity to Go6850 but insensitivity to Go6976. In isolated rat hepatocytes with intact bile secretion, no colocalization of rat isoforms of the bile salt export pump (Bsep) and Golgi was found, but colocalization occurred after inhibition of p38(MAPK) and PKC, suggesting that Bsep trafficking to the canalicular membrane depends on the basal activity of these kinases in polarized cells. CONCLUSIONS: p38(MAPK) regulates BSEP trafficking from the Golgi to the canalicular membrane, and the Golgi may serve as a BSEP pool in certain forms of cholestasis or when p38(MAPK) activity is inhibited. Activation of p38(MAPK) by TUDC can recruit Golgi-associated BSEP in line with its choleretic action. PMID- 14762792 TI - Phosphatidylinositide 3-kinase gamma regulates key pathologic responses to cholecystokinin in pancreatic acinar cells. AB - BACKGROUND & AIMS: Early events in the pancreatic acinar cell critical for development of pancreatitis include activation of the transcription factor nuclear factor kappa B (NF-kappa B), abnormal Ca(2+) responses, and trypsinogen activation. Mechanisms underlying these responses, which can be studied in isolated pancreatic acini stimulated with supraphysiologic doses of cholecystokinin (CCK-8), remain poorly understood. We here report that these responses are regulated by phosphatidylinositide 3-kinase (PI3K) gamma. METHODS: To inactivate PI3K, we used mice deficient in the catalytic PI3K gamma subunit p110 gamma as well as the PI3K inhibitors LY294002 and wortmannin. We measured Ca(2+) responses by using Fura-2, NF-kappa B-binding activity by electromobility shift assay, I kappa B degradation by Western blotting, and trypsinogen activation by fluorogenic assay. RESULTS: CCK-induced intracellular Ca(2+) mobilization, Ca(2+) influx, trypsinogen, and NF-kappa B activation were all diminished in pancreatic acini isolated from p110 gamma(-/-) mice. Both in mouse and rat acini, these responses were inhibited by the PI3K inhibitors. The Ca(2+) signal and trypsinogen activation were similarly reduced in acini isolated from p110 gamma(-/-) and p110 gamma(+/-) mice compared with wild-type mice. By contrast, NF-kappa B activation was inhibited in p110 gamma(-/-) acini but not in p110 gamma(+/-) acini. These differences indicate that the mechanism of NF-kappa B regulation by PI3K gamma differs from those for the Ca(2+) and trypsinogen responses. CCK-induced responses in p110 gamma(-/-) acini were all further inhibited by LY294002, indicating the involvement of other PI3K isoform(s), in addition to PI3K gamma. CONCLUSIONS: The results show that key pathologic responses of the pancreatic acinar cell are regulated by PI3K gamma and suggest an important role for this PI3K isoform in pancreatitis. PMID- 14762793 TI - Intestinal metaplasia at the gastroesophageal junction. PMID- 14762794 TI - Gastrointestinal cancers and neurofibromatosis type 1 features in children with a germline homozygous MLH1 mutation. AB - Heterozygous germline DNA mismatch repair gene mutations are typically associated with hereditary nonpolyposis colorectal cancer. The molecular hallmark of this syndrome is high-frequency microsatellite instability in the tumors. Rare childhood cases with homozygous or compound heterozygous DNA mismatch repair gene mutations have a described predisposition to leukemia, lymphoma, and brain tumors but not to gastrointestinal cancer. We have now characterized a family in which 2 children with a homozygous germline DNA mismatch repair gene mutation developed early-onset gastrointestinal cancers. The 11-year-old proband had cafe-au-lait macules and developed metastatic duodenal adenocarcinoma that arose in a tubulovillous adenoma. His 9-year-old sister with cafe-au-lait macules and axillary freckling presented with malignant colon polyps. A 6-year-old sister with cafe-au-lait macules, hairy nevi, and a plexiform neurofibroma of the tongue has no malignancies to date. The family history did not fulfill the Amsterdam criteria for hereditary nonpolyposis colorectal cancer, but 2 relatives in their 60s had gastric cancer and colorectal cancer, whereas the parents, who are first cousins, remain cancer free. The proband's metastatic duodenal cancer and his sister's malignant colon polyps had high-frequency microsatellite instability but had detectable MLH1, MSH2, and MSH6 proteins by immunohistochemistry. Because some germline DNA mismatch repair gene deficiencies are associated with apparently intact immunohistochemical DNA mismatch repair gene expression in tumors, we proceeded to DNA sequencing, which showed that all 3 children had a germline homozygous MLH1 missense mutation (exon 18, codon 687, CGG-->TGG), whereas both parents were heterozygous for this mutation. PMID- 14762795 TI - Steatosis and hepatitis C virus: mechanisms and significance for hepatic and extrahepatic disease. AB - Nonalcoholic fatty liver disease (NAFLD) and hepatitis C virus (HCV)-related liver disease are common in the general population, but their concurrence is 2- to 3-fold higher than would be expected by chance alone. In patients with chronic HCV infection, steatosis is attributable to a variable combination of the mechanisms considered to play a role in the pathogenesis of NAFLD--insulin resistance in the obese and in the lean subject--along with a direct effect of HCV on hepatic lipid metabolism that leads to triglyceride accumulation through inhibition of export proteins that are required for very low density lipoprotein (VLDL) assembly and secretion. Accumulating evidence suggests that steatosis contributes to the progression of fibrosis in HCV-related disease in a pattern similar to that observed in NAFLD. Potential mechanisms of this effect include the increased sensitivity of steatotic livers to oxidative stress and cytokine mediated injury. Steatosis-related hepatic insulin resistance may also play a role through the profibrogenic effects of the compensatory hyperinsulinemia and provides a potential explanation for the association between HCV and type 2 diabetes mellitus. Indeed, an appreciation of the importance of fat in HCV has recently led to trials of adjuvant therapy for HCV directed at steatosis associated disease mechanisms, with encouraging results reported for various modalities, including weight loss and antioxidants. Future therapy should be aimed at exploiting the interactions of HCV with host insulin and lipid metabolism, particularly in nonresponders to standard antiviral schedules. PMID- 14762796 TI - Episodic retreatment versus scheduled maintenance therapy of crohn's disease with infliximab: not so far apart. PMID- 14762797 TI - Antibody responses in Crohn's disease. PMID- 14762798 TI - What every hepatologist should know about endocrinology: obesity, diabetes, and liver disease. PMID- 14762799 TI - Genetic dissection of the secretory machinery in the stomach. PMID- 14762800 TI - Anti-reflux surgery for Barrett's esophagus? PMID- 14762801 TI - IBD researchers reach for the "tabloid". PMID- 14762802 TI - Molecular hijacking and intracellular chaos. PMID- 14762804 TI - Hepcidin in HFE-associated hemochromatosis: another piece of the "iron" puzzle. PMID- 14762825 TI - Selective alkane C-H-bond functionalizations utilizing oxidative single-electron transfer and organocatalysis. AB - Alkane C-H-bond functionalization methods not utilizing metal-catalysis are discussed based on experimental and computational data, beginning with molecule induced homolysis (reactions of alkanes with dioxiranes). Electrophilic reactions are elaborated next with an emphasis on mechanistic details that reveal that many so-called electrophilic C-H or C-C-bond insertions can be rationalized by electron-transfer reactions (inner sphere, H-coupled, and outer sphere). Finally, radical functionalizations utilizing carbon-centered (relatively stable) radicals generated under organocatalytic (phase-transfer catalysis, PTC) conditions are presented as valuable alternatives to other radical-chain alkane functionalizations. The remarkable chemo- and regioselectivities of these PTC radical reactions and the tolerance of high strain in certain aliphatic hydrocarbons make them particularly useful for laboratory-scale halogenations, in particular, iodinations of unactivated alkane C-H-bonds. PMID- 14762826 TI - Lessons learned from macrolide synthesis. AB - The highlights of three macrolide syntheses recently completed in our laboratory are described. In the epothilone B synthesis we developed the "early epoxide approach," which resulted in the first completely stereocontrolled synthesis of this natural product. In the laulimalide synthesis our contribution was the auxiliary controlled ene-macrocyclization and Sharpless' kinetic resolution for achieving a regioselective epoxidation of two pseudo-enantiomorphic allylic alcohol subunits. The tartrolon B synthesis was the first to be completed. In this case, a substrate controlled aldol addition was used to assemble the entire carbon skeleton of the compound. PMID- 14762827 TI - Metal-catalyzed reactions of diborons for synthesis of organoboron compounds. AB - Metal-catalyzed borylation of alkenes, alkynes, arenes, and organic halides with B-B or H-B compounds has been developed for the synthesis of organoboron compounds from simple organic substrates. The platinum(0)-catalyzed addition of bis(pinacolato)diboron to alkenes and alkynes provided a method for the stereoselective synthesis of cis-bis(boryl)alkanes or cis-bis(boryl)alkenes. The addition of diboron to 1,3-dienes with platinum(0) complexes provided a new access to cis-1,4-bis(boryl)-2-butene derivatives, which are versatile reagents for diastereoselective allylboration of carbonyl compounds. The first one-step procedure for the syntheses of aryl-, vinyl-, and allylboronates was achieved via crosscoupling reactions of diborons with aryl and 1-alkenyl halides or triflates and allyl acetates. Direct C-H borylation of arenes catalyzed by a transition metal complex was studied as an economical protocol for the synthesis of a variety of arylboron derivatives. Ir-catalyzed C-H borylation of arenes, heteroarenes, and benzylic positions of alkylarenes by bis(pinacolato)diboron or pinacolborane furnished aryl-, heteroaryl-, and benzylboron compounds. This article discusses the mechanisms of these reactions and their synthetic applications. PMID- 14762829 TI - Photoswitching of ferroelectric liquid crystals using chiral thioindigo dopants: The development of a photochemical switch hitter. AB - By virtue of its spontaneous polarization (PS), a ferroelectric SmC* liquid crystal can be switched between two states corresponding to opposite molecular tilt orientations using an electric field, thus producing an ON-OFF light shutter between crossed polarizers. Considerable efforts have been made over the past decade to develop photonic FLC light shutters because of their potential uses in dynamic holography and optical data storage. The ON-OFF switching of a FLC light shutter can be triggered by light via a photoinversion of PS using a photochromic dopant. The spontaneous polarization is a chiral bulk property that can be left handed (negative) or right-handed (positive), depending on the absolute configuration of the chiral component of the SmC* phase. In the approach described herein, the magnitude of PS is modulated via the photoisomerization of a chiral thioindigo dopant that undergoes a large increase in transverse dipole moment upon trans-cis photoisomerization. The sign of PS is photoinverted using an "ambidextrous" thioindigo dopant containing a chiral 2-octyloxy side chain that is coupled to the thioindigo core and induces a positive PS, and a chiral 2,3-difluorooctyloxy side chain that is decoupled from the core and induces a negative PS. In the trans form, the 2,3-difluorooctyloxy side chain predominates and the net PS induced by the dopant is negative. However, upon trans-cis photoisomerization, the increase in transverse dipole moment of the 2 octyloxy/thioindigo unit raises its induced PS over that of the decoupled 2,3 difluorooctyloxy side chain, and thus inverts the net sign of PS induced by the dopant from negative to positive. PMID- 14762828 TI - Unique sugar metabolism and novel enzymes of hyperthermophilic archaea. AB - Hyperthermophiles are a group of microorganisms that have their optimum growth temperature above 80 degrees C. More than 60 species of the hyperthermophiles have been isolated from marine and continental volcanic environments. Most hyperthermophiles belong to Archaea, the third domain of life, and are considered to be the most ancient of all extant life forms. Recent studies have revealed the presence of unusual sugar metabolic processes in hyperthermophilic archaea, for example, a modified Embden-Meyerhof pathway, that has so far not been observed in bacteria and eucarya. Several novel enzymes, such as ADP-dependent glucokinase, ADP-dependent phosphofructokinase, glyceraldehyde-3-phosphate ferredoxin oxidoreductase, phosphoenolpyruvate synthase, pyruvate : ferredoxin oxidoreductase, and ADP-forming acetyl-CoA synthetase, have been found to be involved in a modified Embden-Meyerhof pathway of the hyperthermophilic archaeon Pyrococcus furiosus. In addition, a unique mode of ATP regeneration has been postulated to exist in the pathway of P. furiosus. The metabolic design observed in this microorganism might reflect the situation at an early stage of evolution. PMID- 14762830 TI - Differences and similarities in the approach to hepatocellular carcinoma between Eastern and Western institutions. PMID- 14762831 TI - Focus on dysplastic nodules and early hepatocellular carcinoma: an Eastern point of view. AB - Although increasing numbers of equivocal nodular lesions have been detected in patients with liver cirrhosis with the development of various diagnostic imaging modalities, the pathological diagnosis of small, well-differentiated hepatocellular carcinoma (HCC) in the early stage and of high-grade dysplastic nodules (DNs) is a controversial issue among both Japanese and Western pathologists. In particular, many of the vaguely nodular HCCs of well differentiated HCC diagnosed by Japanese pathologists tend to be interpreted as high-grade DNs rather than HCC by Western pathologists. In contrast, many of the high-grade DNs diagnosed by Western pathologists are interpreted as well differentiated HCC by Japanese pathologists. The reasons for the discrepancy between Japanese and Western pathologists can be explained by the following: for Western pathologists, most information comes from the study of HCC and advanced cirrhosis explanted at liver transplantation without detailed clinical information about the nodules; for Japanese pathologists, most information comes from the examination of surgical and biopsy materials together with detailed clinical information that includes meticulous follow-up data on the clinical course of the nodular lesions. To resolve the diagnostic confusion concerning equivocal nodular lesions in the cirrhotic liver, it is necessary to promote the active exchange of clinicopathologic information between Japan and Western countries. PMID- 14762832 TI - Hepatocellular nodules in cirrhosis: focus on diagnostic criteria on liver biopsy. A Western experience. AB - The spectrum of so-called space-occupying small (0.5-2.5 cm) sizable nodules arising in the cirrhotic liver includes a series of hyperplastic (large regenerative), dysplastic (low- and high-grade dysplastic), and malignant hepatocellular (well-differentiated hepatocellular carcinoma, HCC) nodules. Major progress in their classification and understanding was achieved through image analysis techniques and careful histological dissection of explanted native livers. Needless to say, the actual understanding of their natural history is crucial to a proper histological classification. The differential diagnosis of these hepatocellular nodules is difficult, particularly on biopsy specimens of focal liver lesions revealed by ultrasound (US), taken during the follow-up of cirrhotic patients. In this study we attempted to summarize, on the basis of our experience, essential clinicopathological features useful to distinguish the different nodules on needle biopsy. Synoptic tables of differential diagnosis and figures of elementar lesions, which have to be looked for, are provided. Only the continuous integration of clinical features, image analysis information of pathological findings, and follow-up data allows establishing the autonomy of these polymorphic and controversial entities and the boundaries between them. PMID- 14762833 TI - Clinical significance of early hepatocellular carcinoma. AB - Early hepatocellular carcinoma (HCC) is defined as a well-differentiated cancer containing Glisson's triad, but it remains unknown whether this lesion is curable by surgery. We studied 70 patients who had a single HCC smaller than 2 cm in diameter (Stage T1) and who underwent curative hepatectomy and long-term follow up. Based on our typing system, the tumors were assigned as early HCC (n=15), overt HCC (n=52), and non-HCC tumor (n=3). The rate of microscopic regional spread was lower in early HCCs than in overt HCCs (7% vs. 42%; P=.01). After a median follow-up of 6.3 years, both overall survival and recurrence-free survival in the early HCC group were significantly better than those in the overt HCC group (P=.01; P=.001, respectively): the 5-year rates of overall survival were 93% and 54% and those of recurrence-free survival were 47% and 16%, respectively. The early HCC group was at a lower risk of recurrence (relative risk, 0.31; 95% confidence interval, 0.15-0.65; P=.002) and death (0.26; 0.09-0.73; P=.01) than was the overt HCC group. Early HCC is a distinct clinical entity with a high rate of surgical cure. PMID- 14762834 TI - The current status of imaging diagnosis of hepatocellular carcinoma. AB - Early detection of hepatocellular carcinoma (HCC) is important, since the most effective treatment for HCC is surgical resection or local ablation therapy when the tumor is small. Fortunately, recent advances in liver imaging techniques have facilitated the detection of small HCCs. Recent progression of noninvasive imaging technology includes various techniques of harmonic ultrasound (US) imaging with several kinds of US contrast agents, multislice helical computed tomography (CT), and rapid high-quality magnetic resonance (MR) technique with new, tissue-specific contrast agents. These techniques seem to have a strong potential to improve detection and characterization of HCC. In this review, the technique and efficacy of new imaging methods including contrast enhanced US, multislice CT, and recent MR techniques with new contrast agents for detecting and diagnosing HCC will be described. PMID- 14762835 TI - Fine needle biopsy of focal liver lesions: the hepatologist's point of view. AB - Guided biopsy of hepatocellular carcinoma has been recently discussed again due to the progress of imaging techniques and the risk of malignant seeding after the procedure. Ultrasound is probably still the most accurate imaging modality for early detection of nodules arising on cirrhosis, even when compared with more advanced imaging techniques. It can be easily employed in the surveillance of high-risk cirrhotic patients. Ultrasound-guided biopsy has very high sensitivity and almost absolute specificity, which allows the appropriate treatment to start after a positive diagnosis. It also allows correct diagnosis of lymphomatous nodules, the incidence of which is increased in hepatitis C virus-related cirrhosis. The risk of seeding appears limited according to the currently available epidemiological data; this should be considered against the risk of false-positive diagnosis of malignancy based on imaging studies alone. Ultrasound guided biopsy is a valuable tool also for the diagnosis of small nodules (less than 10 mm in diameter). The best accuracy in the sampling of hepatocellular carcinoma nodules is obtained by combining smear cytology and microhistology. This can be achieved by a single biopsy with a fine cutting needle that furnishes pathologic material suitable for both examinations, reducing risks and costs. PMID- 14762836 TI - Indication and contraindication for hepatic resection for liver tumors without fine-needle biopsy: validation and extension of an Eastern approach in a Western community hospital. AB - Fine-needle biopsy (FNB) is associated with problems, such as tumor seeding, which are not negligible. The aim of this study was to validate prospectively the accuracy of our diagnostic work-up without FNB, not just to address but also to rule out from a surgical program patients with focal liver lesions (FLLs). From September 2001 to July 2003, 89 patients were seen at an outpatient clinic for FLLs. Nine patients were excluded because of previous FNB and 18 were excluded because carrier of advanced disease. Sixty-two patients with 101 FLLs were included. Preoperative diagnoses were established by means of clinical histories, serum tumor marker levels, ultrasonography (US), and spiral computed tomography (CT). Other imaging modalities were carried out when it was considered necessary. Forty-eight patients underwent surgery, with histological confirmation of the preoperative diagnosis. The remaining 14 patients underwent a close follow-up. The preoperative diagnoses of 47 of the 48 patients who underwent surgery were confirmed (97.9%). All of the 14 patients ruled out for surgical treatment did not show FLL progression at 6-24 months of follow-up. Of the 9 patients who had FNB previously in other centers, 2 had a wrong histological diagnosis. In view of these results, a diagnostic work-up without FNB seems adequate either to include or to exclude patients with potentially resectable FLL from the surgical program and once more highlight the fact that the use of FNB should be drastically limited. PMID- 14762837 TI - Contrast-enhanced intraoperative ultrasonography in surgery for hepatocellular carcinoma in cirrhosis. AB - Intraoperative ultrasonography (IOUS) is the most accurate diagnostic technique for staging hepatocellular carcinoma (HCC), but has low accuracy in differentiating the new nodules detected in the cirrhotic liver. The aim of this preliminary report is to evaluate whether contrast-enhanced intraoperative ultrasonography (CE-IOUS) could provide additional information to IOUS in patients with HCC. From August 2002 to July 2003, a prospective validation cohort study was conducted. For this purpose, 16 consecutive patients underwent liver resection for HCC using IOUS and CE-IOUS. Intraoperatively, in all patients 4.8 mL of SonoVue was injected intravenously through a peripheral vein. IOUS depicted 16 new focal liver lesions: 10 with no enhancement peculiar to HCC at CE-IOUS pattern and at histology (4) or imaging follow-up (6) proved to be benign; the remaining 6 had enhancement peculiar to HCC and histology confirmed this diagnosis. Two different patterns of enhancement were also recognized at CE-IOUS in those HCC nodules depicted preoperatively: one had no similarity to that observed at CT. CE-IOUS added findings to those of unenhanced IOUS in 50% of patients. These results show that IOUS accuracy and specificity is improved by CE IOUS, with a great impact on surgical strategy and oncological radicality. Furthermore, a wider experience with vascular enhancement patterns with CE-IOUS could provide a new classification for HCC nodules. PMID- 14762838 TI - Hepatectomy for hepatocellular carcinoma: patient selection and postoperative outcome. AB - Hepatic resection and liver transplantation are considered the only curative treatments for hepatocellular carcinoma (HCC). Liver transplantation for HCCs < or =5 cm in diameter has been shown to produce favorable survival results, but its application is limited by the lack of donors. Hepatic resection remains the treatment of choice for patients who are not transplantation candidates because of large tumor, macroscopic vascular invasion, or advanced age. For small HCCs associated with Child's A cirrhosis, hepatic resection should still be considered the first-line treatment, but salvage transplantation for intrahepatic recurrence may be a feasible strategy. Recent improvement in surgical techniques and perioperative care has increased the safety and expanded the indication of hepatic resection for HCC to include large tumors that require extended hepatectomy in cirrhotic patients. Selection of appropriate candidates for hepatectomy depends on careful assessment of the tumor status and liver function reserve. Evaluation of the general fitness of patients is also critical because comorbid illness is an important cause of postoperative mortality, even if the patients have good liver function reserve. With careful patient selection and surgical expertise, the current operative mortality of hepatectomy for HCC is about 5% or less in major centers. Improved long-term survival results after resection of HCC have also been reported recently, with an overall 5-year survival rate of about 50%. The improved perioperative and long-term survival results have strengthened the role of hepatectomy as the mainstay of treatment for HCC despite the availability of a number of other treatment options for localized HCC. PMID- 14762839 TI - The surgical approach to HCC: our progress and results in Japan. AB - Due to the prevalence of hepatitis virus infection, the incidence of hepatocellular carcinoma (HCC) is very high in Japan. Many techniques have been devised by Japanese surgeons to reduce the mortality rate after hepatectomy for HCC: preoperative precise evaluation of hepatic functional reserve, portal venous embolization as preoperative preparation, anatomical and nonanatomical limited resections using intraoperative ultrasonography, and intermittent inflow occlusion during liver transection. Several challenging surgical procedures are also being tried for advanced HCC: HCC with portal and hepatic venous tumor thrombus, multiple and/or recurrent HCC, and HCC in the caudate lobe. As a result, the latest national survey of HCC revealed that operative mortality was 0.9% and the 5-year survival rate after surgery was 52%. Living-donor liver transplantation for adult patients with HCC is another surgical treatment developed in Japan. After the success of adult-to-adult living donor liver transplant using a left liver graft in 1993, a right liver graft, a left liver graft with caudate lobe, and a right lateral sector graft were developed. Indications for reconstructing the middle hepatic vein tributaries in right liver grafts were also proposed. Consequently, in our series of 36 patients with HCC who underwent living-donor liver transplantation, operative mortality was 3%, and the 2-year survival rate was 84%. PMID- 14762840 TI - Pedicle clamping with ischemic preconditioning in liver resection. AB - Hepatic pedicle clamping (HPC) is widely used to control intraoperative bleeding during hepatectomy; intermittent HPC is better tolerated but is associated with blood loss during each period of reperfusion. Recently, it has been shown that ischemic preconditioning (IP) reduces the ischemia-reperfusion damage for up to 30 minutes of continuous clamping in healthy liver. We evaluated the safety of IP for more prolonged periods of continuous clamping in 42 consecutive patients with healthy liver submitted to hepatectomy. IP was used in 21 patients (group A); mean +/- SD of liver ischemia was 54 +/- 19 minutes (range, 27-110; in 7 cases >60 minutes). In the other 21 patients, continuous clamping alone was used (Group B); liver ischemia lasted 36 +/- 14 minutes (range, 13-70; in 2 cases >60 minutes). Two patients in Group A (9.5%) and 3 in Group B (14.2%) received blood transfusions. In spite of the longer duration of ischemia (P=.001), patients with IP had lower aspartate aminotransferase (AST; P=.03) and alanine aminotransferase (ALT; P=not significant) at postoperative day 1, with a similar trend at postoperative day 3. This was reconfirmed by multiple regression analysis, which showed that although postoperative transaminases increased with increasing duration of ischemia and of the operation in both groups, the increases were significantly smaller (P<.001) with the use of preconditioning. In conclusion, the present study confirms that IP is safe and effective for liver resection in healthy liver and is also better tolerated than continuous clamping alone for prolonged periods of ischemia. This technique should be preferred to continuous clamping alone in healthy liver. Additional studies are needed to assess the role of IP in cirrhotic liver and to compare IP with intermittent clamping. PMID- 14762841 TI - Surgical resection of hepatocellular carcinoma. Post-operative outcome and long term results in Europe: an overview. AB - A multicenter retrospective review of 1467 patients treated by liver resection (LR) for hepatocellular carcinoma (HCC) in Europe over a 13-year period showed a mean mortality rate of 10.6%, which was correlated with the extent of LR, the etiology of cirrhosis and the study period with an improvement during the last years. Improved 5-year overall survival (20-51%) and disease-free survival (20 33%) reached similar rates in cirrhotic than in non-cirrhotic patients. Overall results were similar to those reported in Asian series as far as patients and tumor characteristics were comparable. PMID- 14762842 TI - Major liver resections for hepatocellular carcinoma on cirrhosis: early and long term outcomes. AB - Since the lack of donors, liver resections continue to be the treatment of choice for cirrhotic patients with good liver function and resectable hepatocellular carcinoma (HCC). Moreover, over the past 2 decades, an increasing number of major hepatic resections have been performed. The aim of this study is to evaluate short- and long-term outcomes of 55 cirrhotic patients undergoing major hepatic resection with particular attention to the survival of the patients with gross portal vein invasion or large size tumors. Twenty-two patients (40%) required intra- or post-operative blood transfusion. Medium tumor size was 66.6 +/- 29.2 mm; 7 patients had large size (>10 cm) HCCs. A single node was present in 38 cases (69.1%). There was a gross portal vein tumor thrombus (PVTT) in 13 patients (23.6%). Resection was non-curative in 4 cases. In-hospital mortality and morbidity rates were 5.5% and 30.9%, respectively. The overall and disease-free survival rates were 36.2% and 42.8%, respectively. Overall 5-year survival rates of patients with large size tumors was 17.1%. Ten patients with a gross PVTT had an R0 resection with a 26.6% 5-year survival rate. In conclusion, major hepatic resections for HCC can be performed with low mortality and morbidity rates. HCCs with PVTT or greater than 10 cm in size have very limited options of treatment; the favorable long-term results of our study suggest that they should undergo surgery if a radical resection can be achieved. PMID- 14762843 TI - Obesity and diabetes as a risk factor for hepatocellular carcinoma. AB - Ten percent of patients who undergo resection for hepatocellular carcinoma (HCC) associated with chronic liver disease have no detectable cause for this underlying liver disease. Recent studies have shown that patients with cryptogenic chronic liver disease frequently have risk factors for nonalcoholic fatty liver disease (NAFLD). This study examines the incidence of risk factors for NAFLD in patients with chronic liver disease who underwent resection for HCC. Among 210 patients with chronic liver disease who underwent resection for HCC, 18 (8.6%) had no identifiable cause for the underlying liver disease. These patients were assessed for obesity, diabetes mellitus, and histological features of the tumor and the adjacent liver parenchyma. Comparisons were made with matched patients with alcohol- and chronic-viral-hepatitis-related HCC. The prevalence of obesity (50% vs. 17% vs. 14%), diabetes (56% vs. 17% vs. 11%), aspartate aminotransferase/alanine aminotransferase ratio<1 (50% vs. 19% vs. 17%), and steatosis>20% (61% vs. 17% vs. 19%) was significantly higher in patients with cryptogenic liver disease than in patients with alcohol abuse and chronic viral hepatitis (P<0.0001 for each). Well-differentiated tumors were significantly more common in patients with cryptogenic liver disease (89% vs. 64% in patients with alcohol-related HCC vs. 55% in patients with chronic viral hepatitis-related HCC, P<0.0001). In conclusion, the hypothesis that obesity and diabetes mellitus may be important risk factors for cryptogenic chronic liver disease in patients with HCC is supported by the analysis of surgically treated patients. Whether HCC is primarily related to obesity and diabetes mellitus or secondarily to a NAFLD-like parenchymal lesions remains to be clarified. PMID- 14762844 TI - Surgical treatment of hepatocellular carcinoma: similar long-term results despite geographic variations. AB - Recently, the International Cooperative Study Group for hepatocellular carcinoma (HCC) proposed a new staging based on data from multiple centers across the world. The new TNM staging has been shown to be more accurate in the prognostic classification of patients after resection for HCC. This staging is the basis for the new TNM for HCC approved by the AJCC (American Joint Committee on Cancer) and UICC (Union Internationale Contre le Cancer). Although the general applicability of the new staging system has been confirmed, there still remains a marked geographic variation in the clinicopathologic factors of HCC patients based on their country of origin. Tumor size, rates of hepatitis, and degree of underlying liver damage all vary significantly among countries. Despite these geographic variations, recent data reveal similar long-term survival in Western and Eastern centers when these clinicopathologic factors are accounted for. Uniform criteria that account for these clinicopathologic differences need to be developed to assist in stratifying patients across hepatobiliary centers. PMID- 14762845 TI - Liver transplantation in patients with hepatocellular carcinoma. AB - The preferred therapy for hepatocellular carcinoma (HCC) apparently confined to the liver is surgical removal of the tumor. If the location of the tumor and the functional status of the liver are such that resection with an adequate margin can be achieved with low likelihood of subsequent hepatic failure, liver resection is the preferred approach. When HCC apparently localized to the liver is diagnosed in a patient who, by virtue of tumor characteristics or diminished hepatic reserve, is not a candidate for liver resection, liver transplantation becomes a consideration. This work outlines the approach at The Mount Sinai Hospital to the diagnosis, evaluation, preoperative management, transplantation, and posttransplant follow-up in patients with unresectable HCC. The allocation of livers to patients with HCC is reviewed, and predictors of tumor recurrence and results of liver transplantation for HCC are discussed. Finally, the impact of viral hepatitis and of immunosuppression on transplant outcome are discussed. PMID- 14762846 TI - Underestimation of the influence of satellite nodules as a risk factor for post transplantation recurrence in patients with small hepatocellular carcinoma. AB - Liver transplantation offers good results in patients with small hepatocellular carcinoma. However, 3 to 15% of patients still have recurrence, suggesting that factors other than the size and number of nodules are implicated. The aim of our study was to identify predictive factors of recurrence in patients with small hepatocellular carcinoma. Seventy consecutive patients fulfilling Milano criteria and who were transplanted for hepatocellular carcinoma were studied. Forty-six patients had pretransplantation adjuvant local therapy. The size and number of tumors, the clinical and biological characteristics of the patients were recorded before liver transplantation, and histological analysis was performed on the explanted liver. Overall survival rates at 1 and 3 years were 81% and 66%, respectively. Recurrence-free survival rates at 1 and 3 years were 80% and 65%, respectively. Seven patients had tumor recurrence with 1- and 3-year recurrence rates of 5% and 10%, respectively. Satellite nodules on the explanted liver were the only statistically significant predictor of recurrence (P=.0003). None of the patients who did not have satellite nodules had recurrence. There was a significant correlation between satellite nodules and microvascular invasion. Patients with pretransplantation adjuvant therapy had significantly more tumor necrosis, but did not have less satellite nodules. In conclusion, microscopic satellite nodules are a significant predictive factor of tumor recurrence in patients transplanted for small hepatocellular carcinoma. PMID- 14762847 TI - Percutaneous ablation of hepatocellular carcinoma: state-of-the-art. AB - Percutaneous ablation is considered the best treatment option for patients with early-stage hepatocellular carcinoma (HCC) who are not candidates for surgical resection or liver transplantation. Several methods have been developed, including intratumoral injection of ethanol or acetic acid, and thermal ablation with radiofrequency, laser, microwaves, or cryosurgery. Percutaneous ethanol injection (PEI) has been the most widely used technique. Several series have provided indirect evidence that PEI improves the natural history of HCC. Patients with Child-Pugh class A cirrhosis and either a single tumor smaller than 5 cm or as many as three lesions each smaller than 3 cm may achieve a 5-year survival of 50%. The major limitation of PEI is the high local recurrence rate, which may reach 33-43%. Radiofrequency (RF) ablation has emerged as the most powerful alternate method for percutaneous ablation. Recent studies have shown that RF ablation can achieve more effective local tumor control than PEI with fewer treatment sessions. In a randomized trial, local recurrence-free survival rates were significantly higher in patients who received RF ablation than in those treated by PEI, and treatment allocation was confirmed as an independent prognostic factor by multivariate analysis. RF ablation could therefore be considered as the percutaneous treatment of choice for patients with early-stage tumors. Further investigation is warranted to clarify whether current RF technology could offer improved results in patients with intermediate-stage HCC. PMID- 14762848 TI - Multimodal image-guided tailored therapy of early and intermediate hepatocellular carcinoma: long-term survival in the experience of a single radiologic referral center. AB - The best treatment policy for some patients with hepatocellular carcinoma (HCC) and compensated cirrhosis is still controversial. The aim of this study was to evaluate the long-term survival and related prognostic factors of patients with early and intermediate HCC (Liver Unit of Barcelona classification) treated in a radiologic referral center by a multimodal image-guided tailored therapy (MIGTT), applied over time, choosing the procedure patient by patient according to the presentation of the disease. Between May 1996 and May 2003, 374 patients (210 with early and 164 with intermediate HCC) were treated with MIGTT. Radiofrequency ablation was considered the first choice; ethanol injection was preferred for nodules at risk for radiofrequency; and selective chemoembolization was preferred for nodules not recognizable at ultrasound examination, those not retreatable after an unsuccessful ablation technique, or for satellites. The rate of perioperative mortality and major complications was 0.2% and 4.5%, respectively. The 1-, 3-, and 5-year survival rates were 90%, 69%, and 49% and 83%, 43%, and 28% for patients with early and intermediate HCC, respectively. At the univariate analysis, the indicators of a poor prognosis were Child's class B, portal hypertension, abnormal bilirubin, infiltrating tumor, and abnormal serum alpha fetoprotein (AFP) level. In conclusion, within the limits of historical comparisons, in early HCC, MIGTT should be considered an appropriate option for unresectable patients or for resectable patients presenting with adverse prognostic factors. In intermediate HCC, the 3-year survival rate obtained with MIGTT was better than the best survival rate reported with conventional chemoembolization; however, the benefit for patients presenting with poor prognostic factors remains unclear. PMID- 14762849 TI - Hepatic arterial 90Yttrium glass microspheres (Therasphere) for unresectable hepatocellular carcinoma: interim safety and survival data on 65 patients. AB - Hepatocellular carcinoma (HCC) generally arises in a cirrhotic liver and, in most cases, is multifocal and bilobar. Although trans-hepatic artery chemoembolization (TACE) can be highly affective in shrinking tumors, it is limited by virtue of the damage that it can cause to the liver that is already damaged by chronic disease. A high priority in HCC research, after primary prevention and early detection, is to find new treatment modalities that are both effective and non toxic to the underlying cirrhotic liver. A cohort of 65 patients with biopsy proven unresectable HCC have been treated with hepatic arterial 90Yttrium microspheres (Therasphere), and the interim results are reported here. Only 1 cycle of Therasphere treatment ever was performed on 46 patients, 17 patients had 2 cycles, and 2 patients had 3 cycles of therapy. The median dose delivered was 134 Gy, typically as either 5 or 10 GBq (2-4 million microspheres). Clinical toxicities include 9 episodes of abdominal pain and 2 episodes of acute cholecystitis, requiring cholecystectomy. A main lab toxicity was elevated bilirubin which increased by more than 200% in 25 patients (30.5%) during 6 months of therapy, although 18 of these patients had only transient elevation. A prominent finding was prolonged and profound (>70%) lymphopenia in more than 75% of the patients, but without clinical significance. Forty-two patients (64.6%) had a substantial decrease in tumor vascularity in response to therapy, and 25 patients (38.4%) had a partial response, by computed tomography scan. Median survival for Okuda stage I patients (n=42) was 649 days (historical comparison 244) and for Okuda stage II patients (n=23) was 302 days (historical comparison 64 days). All patients were followed after therapy for a minimum of 6 months. There were 42 deaths, 21 due to liver failure, 6 from HCC progression, and 3 from metastases. Therasphere appears to be a relatively safe and effective therapy for advanced-stage unresectable HCC. PMID- 14762850 TI - Prevention and treatment of hepatocellular carcinoma. AB - Viral hepatitis, by either hepatitis C virus (HCV) or hepatitis B virus (HBV), is the dominant cause of hepatocellular carcinoma (HCC). This is to say that HCC may be prevented by controlling viral infection. Horizontal transmission of HCV has become obsolete owing to the discovery of the virus. Vertical transmission of HBV during delivery has been effectively prevented by vaccination and immunization of neonates. The efficacy of interferon therapy against HCV was recently much improved. We now possess several powerful antiviral drugs against HBV. There has been progress also in the treatment of HCC, and together with advances in diagnostics facilitating HCC detection at an early stage, tumor nodules can often be completely removed either by medical ablation or surgical resection. Nevertheless, recurrence of HCC after apparently curative treatment is extraordinarily frequent, since the remaining liver is still at a particularly high risk of HCC. An effective treatment of HCC should include measures to control de novo carcinogenesis. PMID- 14762851 TI - The Barcelona approach: diagnosis, staging, and treatment of hepatocellular carcinoma. AB - Hepatocellular carcinoma (HCC) is the fifth most common neoplasm in the world, and the third most common cause of cancer-related death. It affects mainly patients with cirrhosis of any etiology. Patients with cirrhosis are thus usually included in surveillance plans aiming to achieve early detection and effective treatment. Only patients who would be treated if diagnosed with HCC should undergo surveillance, which is based on ultrasonography and alpha-fetoprotein every 6 months. Upon diagnosis, the patients have to be staged to define tumor extent and liver function impairment. Thereafter, the best treatment option can be indicated and a prognosis estimate can be established. The present manuscript depicts the Barcelona-Clinic Liver Cancer Group diagnostic and treatment strategy. This is based on the analysis of several cohort and randomized controlled studies that have allowed the continuous refinement of treatment indication and application. Surgical resection is considered the first treatment option for early stage patients. It is reserved for patients with solitary tumors without portal hypertension and normal bilirubin. If these conditions are not met, patients are considered for liver transplantation (cadaveric or live donation) or percutaneous ablation if at an early stage (solitary < or =5 cm or up to 3 nodules < or =3 cm). These patients will reach a 5-year survival between 50 and 75%. If patients are diagnosed at an intermediate stage and are still asymptomatic and have preserved liver function, they may benefit from chemoembolization. Their 3-year survival will exceed 50%. There is no effective treatment for patients with advanced disease and thus, in such instances, the patients have to be considered for research trials with new therapeutic options. Finally, patients with end-stage disease should receive only palliative treatment to avoid unnecessary suffering. PMID- 14762852 TI - Post-liver-transplant anemia: etiology and management. AB - Anemia is common after liver transplantation, with the incidence ranging from 4.3% to 28.2% depending on the criteria used to define anemia. The cause of anemia is unidentified in the majority of patients, and it is likely to be multifactorial. Immunosuppressive-medication-induced bone marrow suppression is perhaps the most common cause of unexplained anemia. Chronic blood loss, iron deficiency, hemolysis, and renal insufficiency are other potential causes of chronic anemia. Rare causes, somewhat unique to transplantation, include aplastic anemia, graft-versus-host disease (GVHD), and lymphoproliferative disease. Anemia due to immunosuppressive medication is challenging, since almost all drugs currently used for this purpose cause anemia, but the renal-sparing property of sirolimus may benefit the subgroup in which renal insufficiency is contributing to anemia. Aplastic anemia is seen in young patients transplanted for non-A, non B, non-C, fulminant hepatic failure. It is thought to be immunologically mediated, secondary to an unknown viral infection, and is associated with a grave prognosis. GVHD is another infrequent (approximately 1% of transplant recipients) but serious cause of severe anemia that carries a dismal prognosis. Lymphoproliferative disorder, too may rarely rare cause anemia and it may respond to reduction of immunosuppression. Recipients of solid-organ transplants do not mount a significant increase in erythropoietin in response to anemia. In conclusion, though there are no data on the response of anemia to erythropoietin in liver transplant recipients, it appears to benefit other solid-organ transplant recipients with anemia. PMID- 14762853 TI - Hepatopulmonary syndrome and portopulmonary hypertension: a report of the multicenter liver transplant database. AB - Hepatopulmonary syndrome (HPS) and portopulmonary hypertension (PortoPH) are pulmonary vascular consequences of advanced liver disease associated with significant mortality after orthotopic liver transplantation (OLT). Data from 10 liver transplant centers were collected from 1996 to 2001 that characterized the outcome of patients with either HPS (n = 40) or PortoPH (n = 66) referred for OLT. Key variables (PaO2 for HPS, mean pulmonary artery pressure [MPAP], pulmonary vascular resistance [PVR], and cardiac output [CO] for PortoPH) were analyzed with respect to 3 definitive outcomes (those denied OLT, transplant hospitalization survivors, and transplant hospitalization nonsurvivors). OLT was denied in 8 of 40 patients (20%) with HPS and 30 of 66 patients (45%) with PortoPH. Patients with HPS who were denied OLT had significantly worse PaO2 compared with patients who underwent transplantation (47 vs. 52 mm Hg, P <.005). Transplant hospitalization survival was associated with higher pre-OLT PaO2 (55 vs. 37 mm Hg; P <.005). MPAP was significantly higher (53 vs. 45 mm Hg; P <.015) and PVR was significantly worse (614 vs. 335 dynes. s. cm(-5); P <.05) in patients with PortoPH who were denied OLT compared with patients who underwent transplantation. Transplant hospitalization mortality was 16% (5/32) in patients with HPS and 36% (13/36) in patients with PortoPH. All of the deaths in patients with PortoPH occurred within 18 days of OLT; 5 of the 13 deaths in patients with PortoPH occurred intraoperatively. We concluded that patients with HPS (based on a combination of low PaO2 and nonpulmonary factors) and patients with PortoPH (based on pulmonary hemodynamics) were frequently denied OLT because of pre-OLT test results and comorbidities. For patients who subsequently underwent OLT, transplant hospitalization mortality remained significant for both those with HPS (16%) and PortoPH (36%). PMID- 14762854 TI - Switching monitoring of emulsified cyclosporine from trough level to 2-hour level in stable liver transplant patients. AB - After orthotopic liver transplantation (OLT) many patients use emulsified cyclosporine. Recent data showed that blood levels 2 hours after dosing (C-2) better reflect systemic exposure to the drug (area under the blood concentration time curve) than trough levels (C-0) do. We investigated difference in dosage, creatinine clearance (CrCl), blood pressure (BP), freedom from rejection, and relation of C-2, C-0, and AUC while switching 31 stable patients more than 6 months after OLT from C-0 to C-2 monitoring. With C-0 between 90 and 150 ng/mL we collected 24-hour urine, while blood samples were taken at t = 0, 1, 2, 3, 4, 6, and 8 hours after dosing to measure cyclosporine, creatinine, liver tests, and blood pressure and calculated AUC and CrCl. Target AUC was calculated based on C 0. Then the dose was adjusted to two subsequent C-2 values of 600 ng/mL +/- 15%, the above was repeated, and the differences were assessed. Cyclosporine dose was reduced in 21/31 patients (68%) and remained unchanged in 10/31 patients (32%) after conversion. Mean lowering was 69 mg daily (26.9 %, P < 0.0001). After dose reduction the mean increase of CrCl was 7.93 ml/min (11.6%, P = 0.016). Only systolic and mean morning BP decreased slightly but significantly. C-2 correlated better with AUC0-12 (r2 = 0.75) than C-0 (r2 = 0.64). However, 13/21 patients had a second AUC below target AUC and 2 of these 13 patients developed rejection after conversion to C-2 levels. In conclusion, while C-0 monitoring frequently results in overdosing and more renal dysfunction, C-2 monitoring may lead to episodes of underdosing and rejection. Therefore better ways of monitoring cyclosporine dosing need to be devised. PMID- 14762855 TI - Neoral C2 monitoring in maintenance liver transplant patients: a step forward? PMID- 14762856 TI - Effect of sirolimus on infection incidence in liver transplant recipients. AB - Sirolimus is a new immunosuppressive agent that lacks the nephrotoxicity and neurotoxicity associated with calcineurin inhibitors. The addition of sirolimus to immunosuppressive protocols may thus allow sparing of calcineurin inhibitors and reduction or elimination of associated toxicities. Between January 2000 and July 2001, sirolimus was administered to 55 of 116 consecutive liver recipients. The remaining 61 patients served as the comparison group in the retrospective analysis. In the sirolimus group, perioperative steroids were reduced, and calcineurin inhibitor initiation was delayed. All infectious episodes that occurred within 60 days of liver transplantation were evaluated but were limited to 1 per patient for statistical analysis of sepsis. Demographic variables were comparable between groups. Patients receiving sirolimus experienced more infection (47.2% vs. 18.03%, P<0.001), and this effect persisted across high and low dosage ranges and sirolimus levels. A trend toward increased length of stay was noted (P=0.07). No difference between groups was found in acute rejection rates (17.5% vs. 22.5%), 1-year graft (81% vs. 89%), patient survival (86% vs. 89%), or hepatic artery thrombosis. In conclusion, despite reduction of other immunosuppressants, patients receiving even low doses of sirolimus experienced increased sepsis rates. This agent may have greater usefulness for patients with threatened renal function or patients with chronic rejection after wound healing has occurred. PMID- 14762857 TI - Sustained viral response to interferon and ribavirin in liver transplant recipients with recurrent hepatitis C. AB - Recurrent hepatitis C infection is an important cause of progressive fibrosis, cirrhosis, and graft loss following orthotopic liver transplantation. Treatment for posttransplant recurrence of hepatitis C with interferon-based therapy is difficult but results in loss of detectable virus in up to 30% of patients. However, the durability of viral clearance and the associated histologic response in this setting is unknown. The aim of this study was to determine whether viral loss in response to antiviral therapy is durable and associated with improvement in liver histology. All liver transplant recipients who received interferon-based treatment for recurrent hepatitis C virus (HCV) at the University of Florida from 1991 to 2002 were included in this study. Patients who lost detectable HCV after treatment with interferon alone or in combination with ribavirin were followed to assess the durability of viral response and its impact on liver histology. One hundred nineteen transplant recipients were treated with interferon or combination therapy. Twenty-nine (20 men, 9 women; mean age, 54 yrs [range, 42-74 yrs]) lost detectable HCV RNA and remained virus negative for at least 6 months after discontinuing therapy (sustained viral response[SVR]). The mean follow-up after discontinuing therapy was 24.7 months (range, 6-70 mos). Our study cohort included one patient with SVR following interferon monotherapy and 28 patients with SVR following combination therapy with interferon plus ribavirin. All patients remained HCV RNA negative (assessed by polymerase chain reaction or branched-DNA assay) during follow-up of up to 5 years. Liver histology assessed 2 years after treatment showed less inflammation compared with before treatment in 50% and showed no change in 38%. By 3 to 5 years post-treatment (n = 15 recipients), inflammation was reduced in 60% and remained unchanged in 33%. Fibrosis stage at 2 years improved by > or = 1 stage in 27 %, remained unchanged in 38 %, and worsened in 35% despite viral clearance. At 3 to 5 years, the fibrosis stage had improved in 67%, remained unchanged in 13%, and worsened in 20%. Both grade of inflammation and fibrosis stage improved by 3 to 5 years posttreatment compared with baseline histology (p < 0.05). In conclusion, loss of HCV after treatment of recurrent chronic hepatitis C with interferon and ribavirin is durable, and the durability of the SVR is associated with improvement in hepatic inflammation and regression of fibrosis. PMID- 14762858 TI - E2 quasispecies specificity of hepatitis C virus association with allografts immediately after liver transplantation. AB - It is unknown whether all hepatitis C virus (HCV) quasispecies variants found within patient serum have equal capacity to associate with the liver after transplantation; however, in vitro models of HCV infection suggest that variations in the hypervariable region 1 (HVR1) of the second envelope protein (E2) may be important in infectivity. The hypothesis of the current study is that the two hypervariable regions (HVR1 and HVR2) within E2 are important in the initial virus-liver interaction, and, therefore, certain HCV quasispecies variants will be isolated from the liver after reperfusion. In 8 patients with end-stage liver disease secondary to HCV infection, HCV envelope quasispecies were determined from intraoperative serum samples obtained before the anhepatic phase of transplantation and from liver biopsies 1.5 to 2.5 hours after the transplanted liver was perfused. Explanted (native) liver biopsies were taken as a control. Sequence analysis was performed on clones of specific HCV reverse transcriptase-polymerase chain reaction products spanning HVR1 and HVR2 of the E2 protein. HVR1 was more variable than HVR2 for all samples. Quasispecies isolated from postperfusion liver differed more from serum than did explanted liver quasispecies at HVR1 (P = 0.03) but not at HVR2 (P = 0.2). Comparison of HVR1 sequences from postperfusion liver versus serum revealed significantly less HVR1 genetic complexity and diversity (P = 0.02 and P = 0.04, respectively). Immediately after transplantation but before actual infection, liver allografts select out from the infecting serum inoculum a less heterogeneous, more closely related population of quasispecies variants. PMID- 14762859 TI - Genetic variability of hepatitis C virus NS3 protein in human leukocyte antigen A2 liver transplant recipients with recurrent hepatitis C. AB - The association between the severity of chronic hepatitis C and the variability of the hepatitis C virus (HCV) genome remains controversial, but to our knowledge few data are available to date regarding T-cell epitope coding regions in transplant patients. In the current study, we identified 21 human leukocyte antigen (HLA)-A2-positive Spanish patients with chronic hepatitis C, 14 immunosuppressed liver transplant recipients, and 7 immunocompetent controls. Alanine aminotransferase, aspartate aminotransferase, viral load, and rate of fibrosis progression were determined. Genetic distances of HCV isolates and variations in epitopes of the HCV nonstructural 3 protein (NS3-1393 LIFCHSKKK and NS3-1406 KLVALGINAV) were compared between patients with slow or fast progression of fibrosis. Isolates from transplant patients with fast progression were found to be more divergent (P =.03), had a higher mean value of synonymous (dS) variations (P =.02), and some were differentiated in a phylogenetic tree, compared with isolates from patients with slow progression. The HLA-A2-restricted NS3-1406 epitope was found to be more variable (20 of 21 isolates differed from the prototype) compared with the A3-restricted NS3-1392 epitope (19% vs. 1.25% variation). A shift in the viral peptide was not detected in a subset of transplant patients, but was evident in two of three nontransplant patients with follow-up. There was no correlation noted between a particular amino acid variation and fibrosis progression (slow or fast) in either transplant or nontransplant patients. The results of the current study suggest that 1) there may be different HCV-1b strains in our geographic area, 2) immunosuppression appears to have little effect in amino acid variation at the HCV NS3-1406 epitope, and 3) variations over time might be more frequent in nonimmunosuppressed patients. PMID- 14762860 TI - Psychosocial adjustment to orthotopic liver transplantation in 266 recipients. AB - Although the survival rate of patients undergoing orthotopic liver transplantation (OLT) is highly satisfactory, one of the most important objectives for liver transplantation teams at the present time is to achieve the best possible quality of life and psychosocial functioning for these patients after transplantation. We present the preliminary results of a study designed to determine which domains of psychosocial functioning are most affected in liver transplant recipients, and to examine the factors associated with poorer adjustment after OLT, using a utility-based standardized measure. Patients who had undergone liver transplant more than 12 months previously were eligible. They were administered the Psychosocial Adjustment to Illness Scale (PAIS), and they provided the answers themselves. Multivariate regression models showed that attitudes toward health care were poorer in women (beta = 0.916, P <.001), in patients who were employed at the moment of transplantation (beta = 0.530, P =.032), and in patients of lower social class (beta = 0.722, P =.026) than in men, unemployed patients, and patients of higher social class. Sexual functioning was worse in women (beta = 0.907, P =.001) and older patients (beta = 0.999, P <.001) than in men or younger patients. Psychological distress was higher in women (beta = 0.981, P =.001) than in men, and lower in currently employed patients (beta = -0.937, P =.001) than in the unemployed. Only gender remained significantly associated with the total PAIS score (beta = 0.969, P <.001), with women showing a poorer overall psychosocial adjustment to OLT. In conclusion, there seems to be no doubt that liver transplantation improves quality of life, but special attention should be paid to female recipients, who seem to have more difficulty than their male counterparts in adjusting to the psychosocial consequences of the procedure. PMID- 14762861 TI - Outcome of liver transplantation in adult recipients: influence of neighborhood income, education, and insurance. AB - Poor socioeconomic status (SES) may be associated with lower survival after liver transplantation. In a previous study, we showed that African-American race was an independent predictor of poor survival, and one of the major criticisms of our study was that we had not adjusted the survival for SES as a confounding variable. The objective of the present study was to determine the posttransplant outcome of adult liver transplant recipients based on neighborhood income, education, and insurance using the United Network for Organ Sharing (UNOS) database from 1987 to 2001. Patients (n = 29,481) were divided into 5 groups based on median income as determined by zip code: <30,000 dollars, 30,001-40,000 dollars, 40,001-50,000 dollars, 50,001-60,000 dollars, and >60,000 dollars). Patients (n = 14,814) were divided into 4 groups based on level of education: higher than bachelor's degree; college attendance or technical school; high school education (grades 9-12); less than high school education. Insurance payer status (n = 23,440) was divided into Medicaid, Medicare, government agency, HMO/PPO, and private. Cox regression analysis was used to adjust the survival for other known independent predictors such as age, race, UNOS status, diagnosis, and creatinine. Results showed that neighborhood income had no effect on graft or patient survival either in the entire cohort or within different racial groups. Education had only marginal influence on the outcome; survival was lower in those with a high school education than in those with graduate education. Patients with Medicaid and Medicare had lower survival when compared to those with private insurance. African-Americans had a lower 5-year survival when compared to white Americans after adjusting for SES and other confounding variables. In conclusion, neighborhood income does not influence the outcome of liver transplantation. Education had minimal influence, but patients with Medicare and Medicaid had lower survival compared to those with private insurance. PMID- 14762862 TI - Intermittent inflow occlusion in living liver donors: impact on safety and remnant function. AB - Clamping of the portal triad accomplishes complete inflow occlusion. This maneuver is commonly used during liver surgery to minimize blood loss but is not widely used in living donors undergoing resection for liver transplantation. We compared outcomes in living donors who underwent resection with and without inflow occlusion. We reviewed data on 2 nonsimultaneous living liver donor cohorts. The first 20 donors (group 1) underwent resection without inflow occlusion. In the next 15 donors (group 2), inflow occlusion was used during parenchymal transection, using cycles of 10-15 minutes occlusion and 6 minutes reperfusion. In donors, we recorded type of resection; ischemia times; blood loss; transfusions; peak ALT, AST, bilirubin, and INR in the first 5 days; hospital length of stay (LOS), and major complications. In recipients, we recorded peak ALT. In group 1, 19 of 20 donors underwent right hepatectomy. In group 2, 8 donors underwent right hepatectomy, and 7 donors had left lobectomies. Total ischemic time ranged from 16-49 minutes (mean, 31 +/- 9 minutes). In group 1, two donors received a total of 5 U of allogeneic blood. In group 2, no donor required transfusion. Mean peak ALT was significantly higher in group 1 (521 +/- 336 U/L) than group 2 (322 +/- 162 U/L; P = 0.03). Mean INR was significantly higher in group 1 (1.8 +/- 0.2) vs. group 2 (1.5 +/- 0.2; P = 0.001). There were 4 major complications in group 1 (incisional hernia, transient liver failure, biliary stricture, and biliary leak) and no major intraoperative or postoperative complications in group 2. Mean LOS was significantly longer in group 1 (7.9 +/- 2.9 days) than group 2 (6.2 +/- 1.1 days; P = 0.04). Mean peak ALT in recipients trended lower in group 2. In conclusion, inflow occlusion was associated with reduced blood loss and less ischemic injury to hepatic remnants in the donors and the grafts in the recipients. These benefits were associated with a diminished incidence of major complications and shorter LOS. Inflow occlusion should be an essential part of living donor hepatectomy. PMID- 14762863 TI - 3DCT angiography for detection of vascular complications in pediatric liver transplantation. AB - Catheter angiography for early diagnosis of vascular complications in pediatric liver transplant yields excellent results but remains an extremely invasive examination for younger children, precluding its routine use. We assessed the efficacy of three-dimensional multislice computed tomographic angiography (3DCTA) as an alternative option in these patients. METHODS: Twenty children suspected of vascular complications on clinical grounds, laboratory findings, or Doppler ultrasound underwent 3DCTA between April 2000 and April 2003. Interventional procedures via conventional angiography were subsequently performed in 5 cases, thrombolytic therapy in 4, surgical in 1, and conservative treatment in 10. RESULTS: Two hepatic artery stenosis,1 hepatic artery thrombosis, 5 hepatic vein stenosis, 4 portal vein occlusion, 1 portal vein stenosis, and 7 non-vascular lesions were detected, all of which paralleled the findings of catheter angiography, Doppler ultrasound, and operations. The diagnostic accuracy for vascular complication was 90%. The sensitivity and specificity were 86.7% and 100%, respectively. The positive and negative predictive values were 100% and 71.4%, respectively. To date 19 patients are alive, with a median follow-up period of 24.8 months. In conclusion, 3DCTA is accurate and efficient in the identification of pathological vascular insults and offers essential information for major decision on further management of the vascular complications in pediatric recipients of liver transplant. PMID- 14762864 TI - Porcine partial liver transplantation: a novel model of the "small-for-size" liver graft. AB - Increasing shortage of cadaveric grafts demands the utilization of living donor and split liver grafts. The purpose of this study was to 1) define the "small-for size" graft in a pig liver transplant model 2) evaluate pathological changes associated with small-for-size liver transplantation. Pigs were divided into four groups based on the volume of transplanted liver: (a) control group (n=4), 100% liver volume (LV) (b) group I (n=8), 60% LV (c) group II (n=8), 30% LV (d) group III (n=15), 20% LV. Tacrolimus and methyl prednisone were administered as immunosuppression. Animals were followed for 5 days with daily serum biochemistry, liver biopsies on day 3 and 5 for light microscopy, and tissue levels of thymidine kinase (TK) and ornithine decarboxylase (ODC). Liver grafts were weighed pretransplant and at sacrifice. All the recipients of 100%, 60%, and 30% grafts survived. Transplantation of 20% grafts (group III) resulted in a 47% mortality rate. Group III animals showed significantly prolonged prothrombin times (p<0.05), elevated bilirubin levels (p<0.05), and ascites. The rate of regeneration, as indicated by TK activity and graft weight was inversely proportional to the size of the transplanted graft. The severity of the microvascular injury was inversely proportional to graft size and appeared to be the survival-limiting injury. Frank rupture of the sinusoidal lining, parenchymal hemorrhage, and portal vein injury were prominent in group III animals 1 hour following reperfusion. This study established a reproducible large animal model of partial liver grafting; it defined the small-for-size syndrome in this model and described the associated microvascular injury. PMID- 14762865 TI - Technique for expanding the donor liver pool: heat shock preconditioning in a rat fatty liver model. AB - Fatty liver is a common predisposing risk factor for postoperative liver failure and accounts for most discarded livers during triage of donors. We investigated the effect of heat shock preconditioning (HPc) on recipient survival in a rat fatty liver transplantation model. Fatty liver donor rats were exposed to brief whole-body hyperthermia (10 minutes at 42.5 degrees C) and allowed to recover. HPc induced heat shock proteins (HSPs) (HSP72, HSP90, and heme oxygenase [HO]-1) in donor livers, with levels peaking 12 to 48 hours after HPc. Subsequently, donor livers were harvested 24 hours after HPc, placed in cold storage for 10 hours, and transplanted into normal rats. At 3 hours posttransplantation, HPc reduced serum liver enzymes in the recipients and almost completely suppressed the release of tumor necrosis factor (TNF)-alpha and interleukin (IL)-10. Histologic evaluation 3 and 24 hours after transplantation showed that HPc significantly reduced hepatic inflammation and hepatocellular necrosis without affecting the steatotic appearance of hepatocytes. One week after transplantation, control non-heat-shocked and heat-shocked fatty liver recipients exhibited survival rates of less than 10% and more than 80%, respectively. The evaluation of the survival of recipients receiving fatty livers at different times after HPc showed that the protective effect of HPc was significant when donor livers were transplanted 3 to 48 hours after HPc, with the maximum effect seen 6 to 48 hours after HPc. In conclusion, HPc is a promising avenue to salvage rejected donor fatty livers and enhance the survival rate of fatty liver recipients. We estimate that this technique could increase the annual donor pool by 600 livers. PMID- 14762866 TI - Complement membrane attack complex and hemodynamic changes during human orthotopic liver transplantation. AB - Hemodynamic changes and elevation of intracellular calcium following reperfusion in human liver transplantation occur rapidly and do not match the time course of cytokine expression, therefore, we postulate involvement of other, pre-formed substances, such as complement. We studied 40 adult patients undergoing liver transplantation. Blood was drawn for estimation of C3, C4, C3 degradation product, membrane attack complex, and CH100 levels and elastase (a marker of neutrophil activation) at induction of anesthesia, 5 minutes before reperfusion, 5 minutes and 60 minutes after reperfusion. Cardiac output was measured by thermodilution and systemic vascular resistance was calculated at these same time points. There was a significant rise in C5b-9 membrane attack complex (P =.0012) with a corresponding fall in C3 (P =.0013) and C4 (P =.0002) levels and a rise in C3 degradation product levels (P =.0006). There was no significant change in CH100. These changes very closely followed the hemodynamic changes of a significant fall in systemic vascular resistance index (P =.0024) and increase in cardiac index (P =.0005). Elastase rose from 356 +/- 53 to 557 +/- 40 microg/L (P <.0001). There is complement activation and neutrophil activation at reperfusion in liver transplantation. Dilution alone cannot explain the fall in C3 and C4 levels as there is a corresponding increase in membrane attack complex and C3 degradation product levels with time. As both C3 and C4 are consumed, the classical pathway must be active, though alternative and lectin activated pathways may also be involved. These findings may, at least in part, explain the hemodynamic changes typically seen at reperfusion in liver transplantation. PMID- 14762867 TI - The prophylactic use of tranexamic acid and aprotinin in orthotopic liver transplantation: a comparative study. AB - The efficacy of tranexamic acid (TA) and aprotinin (AP) in reducing blood product requirements in orthotopic liver transplantation (OLT) was compared in a prospective, randomized and double-blind study. One hundred and twenty seven consecutive patients undergoing OLT were enrolled; TA was administered to 64 OLT patients at a dose of 10mg /kg/h and aprotinin was administered to 63 OLT patients at a loading dose of 2 x 10(6) KIU followed by an infusion of 500,000 KIU/h. The portocaval shunt could not be performed in 14 OLT patients in the TA group and in 13 OLT patients in the AP group. However, all OLT patients that received either drug were included in the analysis. Perioperative management was standardized. Hemogram, coagulation tests, and blood product requirements were recorded during OLT and during the first 24 hours. No differences in diagnosis, Child score, preoperative coagulation tests, and intraoperative data were found between groups. No significant differences were observed in hemogram and intraoperative coagulation tests with the exception of activated partial thromboplastin time (aPTT). Similarly, there were no intergroup differences in transfusion requirements. Thromboembolic events, reoperations and mortality were similar in both groups. In conclusion, administration of regular doses of TA and AP during OLT did not result in large differences between the two groups. PMID- 14762868 TI - Antifibrinolytics in liver transplantation: they are effective, but what about the risk-benefit ratio? PMID- 14762869 TI - Thromboelastogram monitoring in the perioperative period of hepatectomy for adult living liver donation. AB - Living donor liver transplantation (LDLT) is becoming a widespread procedure. However, the risk of surgical and medical complications in healthy donors is still a major concern. Hypercoagulability contributes to thromboembolic complications after surgery, but alterations of hemostasis after liver resection are difficult to predict. This study aims to define the perioperative coagulation profile of living liver donors by the use of both routine tests and thromboelastogram (TEG). Ten subjects undergoing right hepatectomy for LDLT were studied. A complete coagulation screening was performed before operation. The coagulation profile was evaluated by platelet count, prothrombin time international normalized ratio (PT-INR), activated partial thromboplastin time (aPTT), and TEG at the beginning and at the end of surgery, and on days 1, 3, 5, and 10 after operation, while the donors were under low molecular weight heparin (LMWH) prophylaxis. At preoperative screening, no subject showed evidence of a prothrombotic state. In all cases, TEG was normal at the beginning of surgery. In the postoperative period, despite decreased platelet counts, increased PT-INR, and normal aPTT values, TEG evidenced the progressive development of hypercoagulability in 4 subjects on day 5 and in 6 subjects on day 10. One donor with a definitely hypercoagulable TEG on day 5 experienced deep venous thrombosis (DVT) on day 8, which was resolved with therapeutic doses of LMWH. In conclusion, despite routine tests suggesting hypocoagulability and LMWH prophylaxis, TEG monitoring showed the unexpected occurrence of hypercoagulability in the majority of the subjects after hepatectomy for LDLT. TEG monitoring could be useful in the perioperative management of donors to guide antithrombotic treatment and increase the safety of the procedure. PMID- 14762870 TI - Fatal disseminated Kaposi's sarcoma following human herpesvirus 8 primary infections in liver-transplant recipients. AB - Human herpesvirus 8 (HHV-8) is associated with the development of Kaposi's sarcoma (KS) and rare lymphoproliferative disorders in immunosuppressed patients. The risk of HHV-8 transmission by liver transplantation and the clinical manifestations of primary infection in this setting have yet to be determined. In order to evaluate this risk, we measured the seroprevalence of HHV-8 among 122 liver donors and their respective recipients before and after transplantation. Molecular methods and immunohistochemical analyses were performed to study the features of HHV-8 infection. Antibodies to HHV-8 were detected in sera of 4 donors before transplantation (3.3%) and of 3 recipients (2.4%). None of the 3 recipients, who were HHV-8 seropositive before transplantation, developed a KS during the follow-up. Four primary HHV-8 infections were detected among the 4 HHV 8 seronegative recipients who received a liver from an HHV-8 positive donor. Among these 4 recipients, 2 particularly immunosuppressed patients developed symptomatic diseases and died a few months after transplantation, harboring disseminated KS and HHV-8 positive lymphoproliferation. In these 2 patients, HHV 8 DNA genome sequences were detectable in peripheral blood mononuclear cells and other tissues with high viremia levels before and at the beginning of HHV-8 related diseases. In conclusion, in liver transplantation recipients, HHV-8 primary infection can be associated with fatal outcome. This study raises the question of screening liver donors for HHV-8--even in low HHV-8 infection prevalence countries--not systematically to exclude the graft but to monitor, clinically and biologically, patients who received a graft from an HHV-8-infected donor. PMID- 14762871 TI - Estimation of glomerular filtration rates before and after orthotopic liver transplantation: evaluation of current equations. AB - The ability to estimate rather than measure the glomerular filtration rate (GFR) in patients before and after liver transplantation would be helpful in estimating risk, dosing drugs, and assessing long-term toxicity of calcineurin inhibitors. Currently available equations for estimating the GFR have not been validated in either the pre- or post-liver transplant population. We have evaluated the performance of currently used formulas for the estimation of the GFR in this setting. Data were collected prospectively on patients who underwent liver transplantation between 1984 and 2001. GFR per 1.73 m2 was measured by I125 iothalamate in patients at the pretransplant evaluation and at 3 months, 1 year, and yearly posttransplant thereafter. GFR estimated by the Cockcroft-Gault equation, the Nankivell equation, and the equations from the Modification of Diet in Renal Disease (MDRD) Study (6, 5, and 4 variables) was compared with the measured GFR. Pretransplant GFR was available in 1,447 patients. The mean GFR was 90.7 +/- 40.5 mL/min. Values for r and r2 were highest for the MDRD Study 6 variable equation (0.70 and 0.49, respectively). Only 66% of estimates were within 30% of the measured GFR. At 3 months, 1 year, and 5 years posttransplant, the mean GFR was 59.5 +/- 27.1 mL/min, 62.7 +/- 27.8 mL/min, and 55.3 +/- 26.1 mL/min, respectively. Values for r and r(2) for the MDRD Study 6-variable equations at 1 and 5 years posttransplant were 0.74 (0.55) and 0.76 (0.58), respectively. At these time points, however, only 67% and 64% of the estimated GFR were within 30% of the measured GFR. MDRD Study equations had greater precision than other equations, but the precision was lower than reported for MDRD estimation of GFR in other populations. Better methods for estimating the GFR are required for evaluation of renal function before and after liver transplantation. PMID- 14762872 TI - Intravascular thrombosis and thromboembolism during liver transplantation: antifibrinolytic therapy implicated? AB - This case report describes a patient who underwent orthotopic liver transplantation and developed extensive hyperacute venous and arterial intravascular thromboses and thromboemboli intraoperatively. The patient was receiving antifibrinolytic therapy with aprotinin. The safety of routine aprotinin therapy in liver transplantation is examined. The value of the thrombelastograph (TEG) as a qualitative assessment of the coagulation system is emphasized. PMID- 14762873 TI - Protective anti-donor IgM production after crossmatch positive liver-kidney transplantation. AB - The mechanism by which a liver transplantation might protect a simultaneous kidney transplant in a crossmatch-positive recipient is unknown. Flow cytometry crossmatch (FCXM) has increased the sensitivity of donor-specific antibody (DSA) detection compared with complement-dependant cytotoxicity (CDC). Here we compare the outcome of a liver-kidney transplantation (LKT), which was CDC and FCXM positive, to the mate-isolated kidney transplantation (KT), which was CDC negative but FCXM positive, from the same donor. Immunoglobulin G (IgG) and immunoglobulin M (IgM) DSAs were measured by FCXM using splenocytes and purified T cells. The KT graft was hyperacutely rejected and removed, but the LKT graft survived without episodes of rejection. Both the KT and the LKT recipients had high levels of circulating antidonor IgG, but not IgM, before transplantation. By day 3, antidonor IgG and IgM in the LKT recipient increased 2 and 7 fold respectively, whereas the KT recipient maintained the high IgG level but did not increase IgM. Histology of the KT graft showed IgG and complement (C1q) deposition, but in the LKT grafts, IgM was deposited without IgG or C1q. Circulating IgG and IgM DSAs returned to background by day 10 and were still at background on day 100. We report a crossmatch-positive LKT where posttransplantation production of IgM DSA, which failed to fix complement, appeared to protect the grafts. PMID- 14762874 TI - De novo sarcoma of donor origin in a liver allograft determined by microsatellite analysis: a short report. AB - This case report describes a patient who underwent liver transplantation for HBV cirrhosis for hepatocellular carcinoma [corrected]. At 3.5 years post transplant, 6 cm tumor was found with CT scanning. With microsatellite analysis it was determined that the tumor was of donor origin. The patient underwent successful right hepatectomy of the tumor that proved to be sarcoma. The patient is doing well without recurrence 1.5 years after resection and 5 years post transplant. PMID- 14762875 TI - Images in liver transplantation. Spontaneous portosystemic shunting several years following liver transplantation: successful treatment via percutaneous embolization. PMID- 14762876 TI - Steroid-free liver transplantation using rabbit antithymocyte globulin and early tacrolimus monotherapy. PMID- 14762878 TI - Predictive values for aspiration after endoscopic laser resections of malignant tumors of the hypopharynx and larynx. AB - BACKGROUND: CO2-laser surgery is a relatively new treatment for selected carcinomas of the upper aerodigestive tract. The purpose of our study was to evaluate prospectively the functional results for swallowing after CO2-laser resections. METHODS: The sample was composed of 210 consecutive patients with malignancies of the larynx and hypopharynx treated with CO2 laser between February 1998 and January 2002. Endoscopic resections included all T1 and T2 tumors and selected T3 and T4 tumors. T1 glottic tumors were not included in the analysis. We assessed the need for a feeding tube and the period the tube remained in place, aspiration pneumonia, tracheotomy secondary to aspiration, the need for a permanent or temporary gastrostomy, and total laryngectomy secondary to aspiration. RESULTS: The nasogastric feeding tube was used in 23.2% of small tumors (2.5 +/- 8.04 days) and in 63% of locally advanced tumors (13.95 +/- 22.55 days). Frequency and period of storage of the feeding tube were higher in locally advanced tumors (p=.0001). Twelve patients (5.7%) had postoperative pneumonia and 59 (28.1%) had temporary postoperative cough during oral intake. Aspiration symptoms correlated with location (p=.001) and locally advanced tumors (p=.016). Eight patients (3.8%) needed a postoperative tracheotomy for severe swallowing difficulties; six (2.9%) of them were definitive and two (0.95%) temporary. Thirteen gastrostomies (6.2%) were performed to avoid severe aspirations; five of them were definitive. The need for gastrostomy correlated significantly with location (p=.002), pT3 and pT4 tumors (p=.002), age (p=.02), and postoperative radiotherapy (p=.04). No correlation was found with the period of feeding tube (p=.38), or aspiration pneumonia (p=.24). CONCLUSIONS: Endoscopic resection of laryngeal and hypopharyngeal tumors is associated with good recovery of deglutition. Many tracheotomies are avoided, the need for a feeding tube is usually reduced, and organ preservation is often feasible even in locally advanced tumors. PMID- 14762879 TI - Single-stage surgical repair of benign laryngotracheal stenosis in adults. AB - BACKGROUND: Benign laryngotracheal stenosis causes considerable morbidity. In a retrospective study, we describe the results of our surgical treatment. METHODS: Between June 1999 and June 2002, 14 adults with laryngotracheal stenosis were referred to our hospital. Stenosis resulted from mechanical ventilation in 11 patients, from Wegener's granulomatosis in 2 patients, and from strangulation in 1 patient. Eleven patients had a tracheotomy. One patient was found unfit for surgery. Nine patients underwent cricotracheal resection (CTR) with end-to-end anastomosis, and four patients underwent single-stage laryngotracheoplasty (SS LTP) without stenting. RESULTS: There were no perioperative deaths. Patients were extubated after mean of 3 days (range, 0-10 days; CTR 2.3 days vs SS-LTP 3.5 days, p=.45). There were in-hospital complications in five patients. Mean hospital stay was 19 days (range, 8-53 days; after CTR 24 days vs SS-LTP 9 days, p=.015). With regard to airway patency and voice recovery, 10 patients (77%) had good results, including 1 patient with two readmissions, and 3 (23%) had satisfactory results, including 1 patient with 11 additional nonsurgical interventions. CONCLUSIONS: Benign laryngotracheal stenosis in the adult patient can be repaired successfully using a strategy of two single-stage surgical procedures. All patients had good or satisfactory functional results. A multidisciplinary approach was essential to achieve these good results. PMID- 14762880 TI - A concurrent chemoirradiation with cisplatin followed by adjuvant chemotherapy with ifosfamide, 5-fluorouracil, and leucovorin for stage IV nasopharyngeal carcinoma. AB - BACKGROUND: To evaluate the toxicity and efficacy of concurrent chemoirradiation with cisplatin followed by adjuvant ifosfamide, 5-fluorouracil and leucovorin in patients with stage IVb nasopharyngeal carcinoma (NPC) PATIENTS AND METHODS: Between October 1998 and August 2001, 35 Chinese patients with stage IVb NPC (N3a:12, N3b:23) were treated with by concurrent chemoirradiation using cisplatin 100 mg/m2 on days 1, 22, and 43 of radiotherapy, followed by adjuvant chemotherapy with 1.4 g/m2, ifosfamide, 450 mg/m2 5-fluorouracil, and 20 mg/m2 leucovorin daily for 5 days and repeated every 3 weeks for three cycles. Radiotherapy was given using standard fractionation at 2 Gy/day to a total of 68 Gy to the nasopharynx and 66 Gy to the neck. RESULTS: All patients completed the prescribed radiotherapy. Twenty-three patients (66%) completed all scheduled cycles of chemotherapy. The compliance rate for concurrent and adjuvant chemotherapy was 71% and 80%, respectively. Grade 3 mucositis occurred in 37%, and grade 3 dermatitis occurred in 11.5% during radiotherapy. Grade 3 neutropenia occurred in 17% during concurrent chemotherapy, and grade 3-4 neutropenia occurred in 48.5% during adjuvant chemotherapy. There were no treatment-related deaths. With a median follow-up of 31 months, the 3-year relapse-free rate was 60%, and the 3-year overall survival rate was 74%. Locoregional control was excellent, with a 3-year local and nodal relapse-free rate of 91% and 83%, respectively. Eleven patients (31%) had developed distant metastases, and the 3 year distant metastasis-free rate was 66%. CONCLUSIONS: The chemotherapy regimen tested is practical with an acceptable compliance rate. Despite having a more advanced stage disease, the observed outcome of our patients seems to be comparable with other series using platinum-based adjuvant chemotherapy. Further investigation to confirm the benefit of using the study regimen in advanced stage NPC is warranted. PMID- 14762881 TI - The treatment of early laryngeal cancers (T1-T2 N0): surgery or irradiation? AB - BACKGROUND: Carcinoma of the larynx is the most common cancer affecting the head and neck region. In Northern Europe, early laryngeal cancer is almost universally treated by irradiation, but elsewhere it is treated by surgery. The main aim of this study was to determine whether there was any difference in survival between the two main therapeutic options. The secondary aim was to assess speech and voice quality in a small, randomized sample of patients from each treatment group. METHODS: The subjects investigated were 488 patients with T1-2, N0 squamous cell carcinoma of the larynx. The patients form an unselected sequential group of our institution's experience with treating this disease over three decades. Four hundred nineteen patients were treated by irradiation, and 69 were treated with surgery. Most surgical patients were treated earlier in the series, whereas radiotherapy later became the treatment of choice. The primary outcome measures were recurrence at the primary site, recurrence in the neck, and tumor specific survival. The secondary outcome measure was speech and voice quality. Statistical analysis was by univariate and multivariate analysis of association and survival. Surgery included horizontal or vertical partial laryngectomy and various minor procedures on the glottis, including cordectomy. Over a 30-year period, radiotherapy was administered to a dose of 60-66 Gy given over 30-33 daily fractions. RESULTS: Surgery tended to be performed early on in the series and radiotherapy thereafter. Surgery was more likely to be carried out for supraglottic disease. These differences apart, the radiotherapy and surgery groups of patients were well matched. The 5-year tumor-specific survival for those treated by irradiation was 87% and for surgery it was 77% (p=.1022). Glottic cancer and T1 disease were associated with high 5-year survivals: 90% and 91%, respectively. Supraglottic site and T2 disease both had a poorer prognoses: 79% and 69%, respectively. The differences for both sets of data were significant. There was no significant difference in primary site recurrence rates for the two treatment modalities, but regional recurrence was higher in the surgery group. Further analysis demonstrated that this was not a function of surgery per se but rather of the unit's policy toward the N0 neck at the time surgery was carried out. Regarding speech and voice quality, radiotherapy was far superior to surgery. All patients in the radiotherapy group but only 3 of 10 in the surgery group were judged to have a good or normal voice (p=.0017). CONCLUSIONS: Both surgery and irradiation are equally effective at treating early laryngeal carcinoma. Speech and voice were highly significantly better in patients treated by irradiation than in those treated by surgery. PMID- 14762882 TI - Prognostic factors in sinonasal tumors involving the anterior skull base. AB - BACKGROUND: Anterior craniofacial resection is a standardized procedure for the treatment of ethmoid and frontal orbital tumors with intracranial invasion. METHODS: A retrospective review of 100 patients with sinonasal tumors involving the anterior skull base who underwent combined craniofacial surgery at the Hospital Central de Asturias. RESULTS: The most frequent pathologic entity was adenocarcinoma (53 cases) and other epithelial tumors (29 cases). Five-year actuarial survival according to the Kaplan-Meier method was 40%. Factors such as involvement of surgical margins, orbital periosteum involvement, frontal sinus invasion, or spread into the dura had no significant effect on survival. Survival, however, was affected by the histologic findings of the tumor (p=.03), brain involvement (p=.04), deep soft tissue involvement of the orbit (p=.003), involvement of the sphenoid sinus (p=.001), previous treatment (p=.05), and postoperative recurrence (p=.0000). Neither the INT staging system nor the UICC system showed statistical prognostic significance. After multivariate analysis and Cox regression analysis, only recurrence after craniofacial resection, involvement of soft tissues of the orbit, and invasion of the sphenoid sinus significantly influenced survival. CONCLUSIONS: Standard staging systems did not show statistical prognostic significance. Only involvement of some critical areas was reliable as predictor of an unfavorable outcome. PMID- 14762883 TI - Tailored endoscopic surgery for the treatment of sinonasal inverted papilloma. AB - BACKGROUND: This retrospective study was designed to evaluate the efficacy of tailored endoscopic surgery. Tailored endoscopic surgery aims at resecting the inverted papilloma completely with a customized surgical approach, especially when an en-bloc excision cannot be comprehensively or routinely achieved because of the immense extent of the tumor. METHODS: Between November 1991 and March 2002, 43 patients with sinonasal inverted papillomas were treated by tailored endoscopic surgery. The average duration of follow-up for this population was 25.3 months (range, 9-150 months). A staging system developed by Krouse was adopted for tumor grading. On the basis of tailored endoscopic surgery, 15 localized lesions and 12 smaller extensive lesions (Krouse stages 1 and 2) were treated by ordinary endoscopic resection, whereas 16 larger extensive lesions (Krouse stages 3 and 4) in which the tumors were immense were subjected to sequential segmental endoscopic surgery (SSES). Seven of these 16 larger extensive lesions combined with endoscopic medial maxillectomy because of extensive encroachment of maxillary sinus antrum. RESULTS: Four patients (9.3%) had residual disease, each requiring one revision surgery. All tumors were successfully resected. No patient required lateral rhinotomy or midfacial degloving procedure. No major complications were encountered in any of the patients. None of the patients had residual disease at the time of this writing. CONCLUSIONS: Tailored endoscopic surgery is a safe and effective treatment that obviates the need for more extensive surgery for the management of inverted papilloma. Proper preoperative evaluations, intraoperative determination of extent and attachment of the tumor, close endoscopic follow-up, and expert application of endoscopic techniques are the keys to the successful use of tailored endoscopic surgery. PMID- 14762884 TI - Radiotherapy alone or combined with surgery for adenoid cystic carcinoma of the head and neck. AB - BACKGROUND: The purpose of this study was to analyze the results of radiotherapy (RT) alone or combined with surgery for adenoid cystic carcinoma. METHODS: Between September 1966 and November 2001, 101 previously untreated patients were treated with curative intent with RT alone or combined with surgery. Follow-up ranged from 0.4 to 30.6 years (median, 6.6 years). All living patients had follow up for at least 1 year. RESULTS: The 5- and 10-year rates of local control were as follows: RT alone, 56% and 43%; surgery and RT, 94% and 91%; and overall, 77% and 69%. Multivariate analysis of local control revealed that T stage (p=.0101) and treatment group (p=.0008) significantly influenced this endpoint. The 5- and 10-year rates of distant metastases-free survival were 80% and 73%. The 5- and 10 year absolute survival rates were as follows: RT alone, 57% and 42%; surgery and RT, 77% and 55%; and overall, 68% and 49%. Multivariate analysis of absolute survival revealed that T stage (p=.0043) and clinical nerve invasion (p=.0011) significantly influenced this endpoint. The 5- and 10-year cause-specific survival rates were as follows: RT alone, 65% and 48%; surgery and RT, 81% and 71%; and overall, 74% and 61%. Multivariate analysis revealed that T stage (p=.0008) and clinical nerve invasion (p=.0005) significantly influenced cause specific survival. CONCLUSIONS: The optimal treatment for patients with adenoid cystic carcinoma is surgery and adjuvant RT. A significant proportion of patients with incompletely resectable disease are cured after RT alone. Improvements in locoregional control are offset, in part, by the relatively high incidence of distant metastases. PMID- 14762885 TI - Prioritizing treatment outcomes: head and neck cancer patients versus nonpatients. AB - BACKGROUND: Treatment decisions in head and neck cancer (HNC) might involve consideration of uncertain tradeoffs of one late effect against another or increasing toxicity or residual impairment for increased chance of survival. Understanding how patients prioritize potential outcomes, as well as whether these preferences are similar to those of nonpatients, is important to informed decision making and treatment planning. METHODS: Two hundred forty-seven newly diagnosed HNC patients from nine institutions and 131 nonpatients rank ordered a set of 12 potential treatment outcomes (eg, cure; being able to swallow; normal voice) from highest (1) to lowest (12). RESULTS: Patients and nonpatients were similar with respect to the three items most frequently ranked in the top three, that is, "being cured of cancer," "living as long as possible," and "having no pain" in that order. In contrast, patients more frequently ranked "cure" (90% vs 80%) and less frequently ranked "no pain" (34% vs 52%) in the top three. CONCLUSIONS: Survival seems to be of paramount importance to both patient and nonpatient groups, overshadowing associated toxicities and potential dysfunction. At the same time, patients might be more willing than nonpatients to undergo aggressive treatments and endure acute distress in the interest of potential long term gains (ie, cure or longer survival). PMID- 14762886 TI - Prostatic adenocarcinoma metastatic to the head and neck and the workup of an unknown epithelioid neoplasm. AB - BACKGROUND: Epithelioid tumors in the head and neck are common and include both primary and metastatic lesions. For metastatic lesions, clinical factors, tumor location, and ancillary immunohistochemical studies must be taken into consideration to help the clinician and the pathologist determine the site of origin. One unusual, but important, primary tumor that can metastasize to the head and neck is carcinoma of the prostate (CAP). METHODS: The files of the University of Pennsylvania Department of Pathology were searched for cases of metastatic CAP. All slides were examined, and clinical information was obtained from the referring physician's patient charts. RESULTS: We describe 14 cases of metastatic CAP to the head and neck. Six patients had no history of CAP at the time of biopsy of the head and neck metastasis, and only eight patients had other widespread metastatic disease. Histologically, most of the tumors had epithelioid cells with prominent nucleoli and cribriform, solid, or infiltrating single cell growth patterns. PSA and PSAP immunohistochemical stains were positive in all cases. Seven of 12 patients with known follow-up are alive after radiation or hormonal therapy. CONCLUSIONS: These results demonstrate a fair prognosis with possible prolonged survival with metastatic CAP to the head and neck after appropriate diagnosis and subsequent hormone and radiation therapy. Given this survival advantage with treatment, it is critical to consider the diagnosis of metastatic CAP when evaluating a metastatic malignant epithelioid tumor in the head and neck of an elderly man. PMID- 14762887 TI - Technique of endoscopic retrograde puncture and dilatation of total esophageal stenosis in patients with radiation-induced strictures. AB - BACKGROUND: Complete esophageal stenosis can occur after external beam radiation therapy for malignancies. Treatment for this complication has traditionally involved surgery. METHODS: A new technique to reestablish luminal patency is described. This minimally invasive technique involves retrograde endoscopy by means of gastrostomy tube tract and puncture of the stenotic occlusion followed by stricture dilatation. The procedure is performed under combined endoscopic and laryngoscopic guidance. RESULTS: Five consecutive patients who had complete esophageal stenoses develop after radiation therapy for malignant disease underwent retrograde endoscopy by way of gastrostomy tube tracts. Stenoses were punctured under endoscopic and laryngoscopic guidance with guide wires. Strictures were dilated with wire-guided balloons or polyvinyl dilators. Luminal patency was established in all patients using this technique without procedural complications. CONCLUSIONS: Endoscopic retrograde puncture and dilatation of total esophageal stenoses is safe, effective, and useful to reestablish luminal patency for radiation-induced strictures. This technique should be attempted before more invasive treatments. PMID- 14762888 TI - Verrucous carcinoma of the paranasal sinuses: case report and clinical update. AB - BACKGROUND: Verrucous carcinoma is a low-grade malignancy that has been reported to occur in all anatomic sites of the head and neck. Fourteen cases of verrucous carcinoma of paranasal sinus origin have been reported to date in the English literature. METHODS: Case report and retrospective review of all cases of verrucous carcinoma of the paranasal sinuses in the English literature. All authors were contacted to provide missing data and long-term follow-up. RESULTS: Five of the eight authors contacted responded, and the most current data from all 15 cases was compiled. Eleven of the 15 patients (73%) were men and ranged in age from 35 to 81 years (median, 68 years). The maxillary sinus is the most common paranasal sinus involved (93%). Presentation often occurred late, with 12 of 15 (80%) initially being seen at stage T3 or higher. Surgical excision was the treatment of choice, and median disease-free survival was 54 months. CONCLUSIONS: Verrucous carcinoma of the paranasal sinuses is a rare but potentially curable disease. Treatment is surgical, and prognosis is good with early intervention. PMID- 14762889 TI - Parapharyngeal lymph node metastasis: an unusual presentation of papillary thyroid carcinoma. AB - BACKGROUND: Parapharyngeal space nodal metastases are usually secondary to malignancies of the pharynx and sinonasal tract, although localization of lymphomas is also possible. Parapharyngeal metastases arising from thyroid papillary carcinoma are instead an exceedingly rare event, with only 10 cases reported up to now in the literature. METHODS: We describe two cases of parapharyngeal metastasis from thyroid papillary carcinoma in a man and a woman, aged 40 and 52 years, respectively. RESULTS: Both patients had a lesion that clinically appeared to be located in the parapharyngeal space; they underwent CT and MRI, which detected a cystic mass in the poststyloid compartment. In the first patient, fine-needle aspiration cytology failed in identifying the histologic nature of the lesion, which was excised through a transcervical approach. A diagnosis of metastatic thyroid papillary carcinoma was rendered and therefore the patient underwent total thyroidectomy. In the second patient, a total thyroidectomy, previously scheduled for multinodular goiter, was performed along with the removal of the parapharyngeal mass. Definitive histologic findings revealed that the two parapharyngeal masses were cystic metastases from a thyroid papillary carcinoma. Both patients received postoperative 131I treatment. Twenty four months after surgery, the first patient is free of disease, whereas the second one has clear signs of abnormal 131I uptake in the lungs. CONCLUSIONS: The differential diagnosis of a parapharyngeal poststyloid mass should also include metastasis from thyroid papillary carcinoma. When the lesion displays a cystic appearance on imaging, it is advisable to rule out a thyroid primary by ultrasonographic examination. The occurrence of a metastasis in such unusual site, even though rarely reported, does not seem to significantly affect the prognosis of the disease. PMID- 14762890 TI - A case of radiation recall mucositis associated with docetaxel. AB - BACKGROUND: Radiation recall reactions, in particular dermatitis, are well documented in the literature. However, radiation recall mucositis is a rare clinical phenomenon. METHODS: We report a case of a 45-year-old man diagnosed with squamous cell carcinoma of the base of tongue. He was treated with surgery followed by chemotherapy and radiation therapy. Several months after completing treatment, he had a recurrence develop outside of the previously irradiated field. He was offered radiation therapy concurrent with docetaxel as salvage therapy. RESULTS: During salvage therapy, acute recall mucositis developed corresponding to his previously irradiated fields. His chemotherapy with docetaxel was withheld, and his symptoms rapidly improved. CONCLUSIONS: This case describes radiation recall mucositis associated with docetaxel, a rare but potentially serious clinical situation. Given the potential severity of the reaction and increasing use of docetaxel as second-line treatment of recurrent head and neck cancers, it is important to be aware of this phenomenon. PMID- 14762891 TI - Polymorphism of NCX4016, an NO-releasing derivative of acetylsalicylic acid. AB - NCX4016 [2-acetoxybenzoic acid 3'-(nitrooxymethyl)phenyl ester] is a recently developed nitrooxy-derivative of aspirin with improved antiinflammatory, analgesic, and antithrombotic activity as well as increased gastrointestinal safety. Systematic polymorphic screening performed with different solvents and preparation methods resulted in the identification of two polymorphs, designated Forms I and II. They were characterized by scanning electron microscopy, powder X ray diffraction, thermal analyses, and infrared spectroscopy; the crystal structure of polymorph I was solved by single-crystal X-ray analysis and compared with that of aspirin. Finally, intrinsic dissolution rate studies and calculations according to the melting data method were performed to assess the thermodynamic relationship between the two polymorphs. PMID- 14762892 TI - Degradation of NSC-281612 (4-[bis[2-[(methylsulfonyl)oxy]ethyl]amino]-2-methyl benzaldehyde), an experimental antineoplastic agent: effects of pH, solvent composition, (SBE)7m-beta-CD, and HP-beta-CD on stability. AB - NSC-281612 (4-[bis[2-[(methylsulfonyl)oxy]ethyl]amino]-2-methyl-benzaldehyde, 1), is a chemically unstable, poorly water soluble, experimental antineoplastic agent. The saturated solubility in water at 25 degrees C was determined as approximately 30 microg/mL. In the pH range 2-11, 1 displayed pH-independent stability (t(50) was around 24 hr). However, an increase in the degradation rate was observed at pH 12. The hydrolysis of the methane sulfonate groups to the corresponding hydroxyl groups was the major degradation pathway in water in the absence of buffers and added halide ions. In phosphate buffer solutions without sodium chloride, phosphate degradants appear to be formed in addition to the mono and dihydroxy degradants. Additional degradants, the mono- and dichloro degradation products, were formed when the ionic strength of the solution was adjusted with sodium chloride. When bromide and iodide ions were added, the corresponding mono- and dihalides were formed. The chloro compounds subsequently underwent further degradation to the hydroxy products. A deuterium kinetic solvent isotope effect study showed that water was minimally involved in the rate determining step. The addition of either (SBE)(7m)-beta-cyclodextrin (CD) or HP beta-CD resulted in a significant enhancement in drug solubility and stability. The apparent binding constants for HP-beta-CD and (SBE)(7m)-beta-CD were 1,486 and 2,740 M(-1), respectively. The stability of 1 in the presence of 0.1 M HP beta-CD and (SBE)(7m)-beta-CD was enhanced 9- and 15-fold, respectively. Thus, (SBE)(7m)-beta-CD displayed better solubilization and stabilization efficacy than HP-beta-CD. PMID- 14762893 TI - In vitro nimesulide absorption from different formulations. AB - In light of improving the bioavailability of poorly water-soluble drugs, this work focused on the comparison among different nimesulide formulations resorting to in vitro absorption experiments through everted rat intestine. The performance of a nimesulide ethanol-triacetin solution, an activated system made up by cogrinding nimesulide/polyvinylpyrrolidone and simple solid nimesulide were compared with that of a reference nimesulide solution. Although ethanol-triacetin solution showed a better performance than the solid nimesulide because wettability problems connected with nimesulide were completely zeroed, the activated system showed a better performance than the reference solution one. This was due to the fact that the activated system allowed to overcome both the wettability and solubility problems connected with nimesulide. Moreover, as proved by intestinal pictures taken before and after permeation experiments, we observed the adhesion of polymeric particles to intestinal villi, this giving origin to a thin layer, surrounding the intestine, characterized by a nimesulide concentration higher than that in the release environment bulk. A proper mathematical model, based on Fick's second law, was developed to model drug absorption in the case of solution and activated system. In this manner, we could calculate nimesulide permeability through the intestinal wall, and we could better define the nature of the above-mentioned thin layer surrounding the intestine. Finally, the mathematical model was used to verify the theoretical correctness of the widely employed technique consisting in data correction for dilution when sample withdrawal and replacement were needed to measure drug concentration in the receiver environment. PMID- 14762894 TI - Extension of the isobolographic approach to interactions studies between more than two drugs: illustration with the convulsant interaction between pefloxacin, norfloxacin, and theophylline in rats. AB - This work proposes a model to characterize the additivity or the nonadditivity of combinations of more than two agents. Using a Bayesian framework, we modeled the variability between experimental subjects, the errors that occurred during data collection, and the relationship between effects and concentrations of agents at the effect site. The model was used to characterize the additivity (or non additivity) of norfloxacin, pefloxacin, and theophylline in causing maximal seizures in male Sprague Dawley rats. Animals received the drugs separately or in various combinations. Drug infusion was stopped at the onset of maximal seizures, and cerebrospinal fluid samples were collected for determination of drug concentration by high-performance liquid chromatography. The model was fitted to concentration data using Markov Chain Monte Carlo techniques. Results showed that induction of seizures by mixtures of theophylline and pefloxacin were additive. Seizure induction by mixtures of norfloxacin and pefloxacin or norfloxacin and theophylline were not additive and, given the model, these drugs interacted negatively. There was no triple interaction effect between the drugs. This study demonstrates the ease with which mixtures of more than two drugs can be analyzed with the proposed model. PMID- 14762895 TI - Ritonavir-PEG 8000 amorphous solid dispersions: in vitro and in vivo evaluations. AB - Ritonavir is a large, lipophilic molecule that is practically insoluble in aqueous media and exhibits an exceedingly slow intrinsic dissolution rate. Although it has favorable lipophilicity, in vitro permeability studies have shown that ritonavir is a substrate of P-glycoprotein. Thus, the oral absorption of ritonavir could be limited by both dissolution and permeability, thereby making it a Class IV compound in the Biopharmaceutics Classification System. Because formulations rarely exert direct influence on local intestinal permeability, the effect of enhanced dissolution rate on oral absorption was explored. More specifically, poly(ethylene glycol) (PEG)-amorphous ritonavir solid dispersions were prepared with different drug loadings, and the in vitro and in vivo performances of the dispersions were evaluated. In vitro dissolution was conducted in 0.1N HCl with a USP Apparatus I. A crossover design was used to evaluate the oral bioavailability of amorphous dispersions relative to crystalline drug in beagle dogs. Intrinsic dissolution measurements of the two solid phases indicated a 10-fold improvement in intrinsic dissolution rate for amorphous ritonavir compared with the crystalline counterpart. In vitro dissolution of ritonavir depended on the solid phase as well as drug loading of the dispersion. In vivo study results indicate that amorphous solid dispersions containing 10-30% drug exhibited significant increases in area under the curve of concentration versus time (AUC) and maximum concentration (C(max)) over crystalline drug. For example, 10% amorphous dispersion exhibited increases of 22 and 13.7-fold in AUC and C(max), respectively. However, both in vitro dissolution and bioavailability decreased with increasing drug load, which led to the construction of a multiple Level C in vitro-in vivo relationship for this Class IV compound. The established relationship between in vitro dissolution and in vivo absorption can help guide formulation development. PMID- 14762896 TI - Interpretation and optimization of the dissolution specifications for a modified release product with an in vivo-in vitro correlation (IVIVC). AB - This article considers the in vivo significance attached to in vitro dissolution testing. Almost invariably, the in vitro dissolution test is interpreted in terms of bioequivalence. The literature that describes methods for setting in vitro dissolution specifications is reviewed. The most common interpretation of these specifications is a deterministic one, that is, those batches passing the dissolution specifications would be bioequivalent with the reference if tested in vivo and those failing the dissolution specifications would not be bioequivalent if tested in vivo. Due to random variation, the deterministic interpretation is not appropriate. Instead, we need to consider the conditional probability that a batch that has passed the in vitro dissolution test would demonstrate bioequivalence if tested in vivo, and that a batch known to have failed the in vitro dissolution test would demonstrate bioinequivalence if tested in vivo. One way to estimate these probabilities is by means of a simulated experiment in which the production and testing (in vivo and in vitro) of a large number of batches is computer simulated. Such a simulation can only be performed if the relationship between the in vitro dissolution characteristics and the in vivo performance of the product has been modeled. These models are generally referred to as in vivo-in vitro correlations (IVIVC). The results of one such experiment are described. The above-mentioned conditional probabilities are shown to depend on the choice of dissolution specifications. This result leads to the notion of optimal dissolution specifications. However, both of the conditional probabilities cannot be maximized simultaneously. The probability of making a correct decision on the basis of the in vitro dissolution test is introduced as a possible optimality criterion. This probability is a linear combination of the two conditional probabilities of interest. Using this criterion, the optimal dissolution specifications can be found by searching over the multidimensional space defined by the half width of each interval used in the specifications to find the combination that maximizes this probability. This process is demonstrated using the Nelder-Mead search routine. The choice of dissolution specifications has profound implications for the routine production of the product because if the specifications were very narrow the probability of a batch passing would be low, resulting in a low hit rate. The same computer program used to perform the simulation experiment can be used to estimate the hit rate. Furthermore, it can be used to explore the magnitude of changes required in the parameters describing the test product (particularly variability) to increase a low hit rate to an acceptable level. PMID- 14762897 TI - Effect of relative humidity on the photocatalytic activity of titanium dioxide and photostability of famotidine. AB - Titanium dioxide (TiO2) has been widely used as a pharmaceutical excipient and is also known to be a strong photocatalyst. An investigation into the relationship between the photocatalytic activity of TiO2 and the photostability of famotidine, which is known as an H2-blocker, is presented. The photocatalytic activity of the anatase form of TiO2, as measured by the four-probe method, is approximately 1.5 times higher than that of the rutile form. Discoloration of famotidine in a binary system containing TiO2 depends significantly on both the wavelength of the irradiating light and the crystal form of the TiO2, with the degree of discoloration of anatase higher than that of the rutile form. Discoloration of famotidine also depends on relative humidity. The relationship between discoloration rate constant and water vapor pressure is linear. These results demonstrate that famotidine is easily discolored by the photocatalytic activity of TiO2 and suggest that the solid-state photocatalytic activity of TiO2 is strongly affected by relative humidity. PMID- 14762898 TI - Influence of feeding and analytical method on the bioequivalence of a racemic drug undergoing enantioselective enterohepatic recycling. AB - Two crossover bioequivalence trials of an enantioselectively enterohepatic recycled drug, carprofen, were conducted in dogs with the same racemic oral formulation to determine: (i) the influence of feeding patterns, and (ii) the effect of the analytical method (enantioselective vs non-enantioselective) on the statistical power of the trials. The first trial was conducted with a standard feeding protocol and the second with a special feeding protocol selected to ensure constant biliary flow into the duodenum. Using a non-enantioselective technique, 90% confidence intervals provided conclusions of bioequivalence in 100% of the cases for both area under the plasma concentration versus time curve (AUC) and maximum plasma drug concentration (Cmax) with the special feeding protocol, but only 50% for AUC and 13% for Cmax with the standard feeding protocol, suggesting that a feeding pattern that diminishes plasma drug concentration rebound for an enterohepatically recycled drug increases the power of a bioequivalence trial. Whatever the feeding protocol, an enantioselective method decreased the power of the trials for AUC but increased the power of the trials for Cmax. For an enterohepatically recycled drug, feeding pattern can influence the power of a bioequivalence trial, and the analytical technique that provides the greatest power depends on the assessed bioequivalence parameter and the feeding pattern. PMID- 14762899 TI - Preparation and crystal characterization of a polymorph, a monohydrate, and an ethyl acetate solvate of the antifungal fluconazole. AB - The preparation and solid-state characterization of three crystalline modifications of the antifungal agent fluconazole [2-(2,4-difluorophenyl)-1,3-bis (1H-124-triazol-1-yl)-propan-2-ol] are reported. Recrystallization of fluconazole from propan-2-ol yielded a polymorph (Form III), whereas the solvents water and ethyl acetate yielded the solvated products fluconazole monohydrate and fluconazole. (ethyl acetate)(0.25), respectively. These species were analyzed by thermogravimetry (TGA), differential scanning calorimetry (DSC), FTIR spectroscopy, powder X-ray diffractometry (PXRD), and single crystal X-ray diffraction. Availability of the hitherto unknown crystal structures facilitated interpretation of the thermal data and clarified previous findings relating to the polymorphism of this compound. Fluconazole was found to exist as a centrosymmetric hydrogen bonded dimer in Form III. For the solvated phases, the solvent locations within the drug host matrices were established as isolated sites for water molecules and constricted channels for ethyl acetate molecules. Desolvation of the monohydrate and ethyl acetate solvate yielded polymorphic Form I. Reference PXRD patterns computed from the refined single-crystal X-ray data for the title compounds are presented. PMID- 14762900 TI - Enhanced oral bioavailability of poorly absorbed drugs. I. Screening of absorption carrier for the ceftriaxone complex. AB - In this study, we attempted to improve the oral absorption of ceftriaxone (CTX) by using an absorption carrier and the CTX complex together. After the CTX-Ca carrageenan gel complex was prepared, several kinds of compounds (Capmul MCM C10, Gelucire 44/14, glycyrrhizin) were screened as potential oral enhancers for our experiment and the intestinal morphologies in rats were examined. Of these compounds, the mono- and diglyceride mixtures, Capmul MCM C10 greatly enhanced the gastrointestinal absorption of CTX when this carrier was coadministered with the complex in rats. Percent bioavailability was attained in rats by the enteral route ranging from 55 to 79% when this complex was administered with various doses of Capmul MCM C10. Furthermore, the surface morphology of the complex has a highly smooth appearance as seen under scanning electron microscopy after preparation. No treatment-related signs of any damage were observed on the administrations intestinal membrane when morphologies were examined in rats after the complex and the absorption carrier were coadministered. The results of the observation suggest that Capmul MCM C10 is a promising carrier, having a good balance between bioavailability enhancing activity and safety, for the oral delivery of CTX when it is coadministered with complex. PMID- 14762901 TI - Image analysis of the shape of granulated powder grains. AB - This study presents and evaluates two new form factors for the characterization of pharmaceutical microparticles using image analysis techniques. The first factor, denoted Vr, is mean percentage variation in radial chord length (for a large number of radial chords drawn at small angular intervals) with respect to mean radial chord length. The second factor, denoted Vp, is percentage deviation of measured perimeter from the perimeter of a circle with radius equal to the mean radial chord length of the particle. Considering both ideal shapes and real pharmaceutical particle populations, these factors are compared with other form factors widely used in pharmaceutical technology. Our results indicate that Vr and Vp allow effective assessment of whether the particles of a given population show pharmaceutically significant deviations from sphericity. The two factors additionally facilitate identification of the basic shapes of particle outlines (notably ellipsoid, rectangular, and irregular). These factors may thus be of value for the characterization and monitoring of pharmaceutical pelleting processes. PMID- 14762902 TI - Measurement of low-dose active pharmaceutical ingredient in a pharmaceutical blend using frequency-domain photon migration. AB - Frequency-domain photon migration (FDPM) measurements of time-dependent light propagation are conducted to provide the powder absorbance for quantitative prediction of terazosin as the active pharmaceutical ingredient (API) in a low dose (0.72 wt %) oral tablet formulation. Calibration of the FDPM-derived powder absorbance at discrete wavelengths of 514, 650, 687, and 785 nm was performed for API contents ranging between 0 and 1.5 wt % in mixtures showing maximum sensitivity at 650 nm. The relative standard deviation (RSD) of FDPM absorption coefficient measurement at 650 nm in a well-mixed 1.08 wt % terazosin blend was <1.6%, of which no more than 0.12% arose from FDPM instrumental error and the remainder was attributable to the complete-random-mixture model. The applicability of FDPM as an on-line sensor for powder-blending operations was further evaluated by analyzing grab samples taken directly from five locations of a 2-cu-ft Gallay blender at intervals of 5 min within the blending process. FDPM results indicate that homogeneity was largely achieved in the first 10 min, during which the RSD of API content across five sampling locations decreased from 27% to 8%, and the RSD decreased to 5% after 25 min of blending. Evolution of homogeneity within the blending process assessed through FDPM measurements was fit to the first-order model of particle blending further evidencing applicability for monitoring powder-blending processes. PMID- 14762903 TI - A novel method for deriving true density of pharmaceutical solids including hydrates and water-containing powders. AB - True density is commonly measured using helium pycnometry. However, most water containing powders, for example, hydrates, amorphous drugs and excipients, and most tablet formulations, release water when exposed to a dry helium atmosphere. Because released water brings significant errors to the measured true density and drying alters the nature of water-containing solids, the helium pycnometry is not suitable for those substances. To overcome this problem, a novel method has been developed to accurately calculate powder true density from compaction data. No drying treatment of powder samples is required. Consequently, the true density thus obtained is relevant to tableting characterization studies because no alteration to the solid is induced by drying. This method involves nonlinear regression of compaction pressure-tablet density data based on a modified Heckel equation. When true density values of water-free powders derived by this novel method were plotted against values measured using pycnometry, a regression line with slope close to unity and intercept close to zero was obtained. Thus, the validity of this method was supported. Using this new method, it was further demonstrated that helium pycnometry always overestimates true densities of water containing powders, for example, hydrates, microcrystalline cellulose (MCC), and tablet formulations. The calculated true densities of powders were the same for different particle shapes and sizes of each material. This further suggests that true density values calculated using this novel method are characteristic of given materials and independent of particulate properties. PMID- 14762904 TI - Thermodynamics of sublimation, crystal lattice energies, and crystal structures of racemates and enantiomers: (+)- and (+/-)-ibuprofen. AB - Thermodynamic differences between ibuprofen (IBP) racemate and the (+)-enantiomer were studied by X-ray diffraction, thermoanalysis, and crystal energy calculations. The thermodynamic functions of sublimation (as a measure of crystal lattice energy) were obtained by the transpiration method. The sublimation enthalpies (DeltaH(sub)) of (+/-)-IBP and (+)-IBP are 115.8 +/- 0.6 and 107.4 +/- 0.5 kJ. mol(-1), respectively. Using the temperature dependency of the saturated vapor pressure, the relative fractions of enthalpy and entropy of the sublimation process were calculated, and the sublimation process for both the racemate and the enantiomer was found to be enthalpy driven (62%). Two different force fields, Mayo et al. (M) and Gavezzotti (G), were used for comparative analysis of crystal lattice energies. Both force fields revealed that the van der Waals term contributes more to the packing energy in (+)-IBP than in (+/-)-IBP. The hydrogen bonding energy, however, contributes at 29.7 and 32.3% to the total crystal lattice energy in (+)-IBP and (+/-)-IBP (M), respectively. Furthermore, different structure fragments of the IBP molecule were analyzed with respect to their contribution to nonbonded van der Waals interactions. The effect of the C-H distance on the van der Waals term of the crystal lattice energy was also studied. PMID- 14762905 TI - Rationalizing the structural properties of bupivacaine base--a local anesthetic- directly from powder X-ray diffraction data. AB - Bupivacaine belongs to a family of 1-alkyl-2',6'-pipecoloxylidides, which has shown promise as reversible action potential blockers that can introduce prolonged local anesthetic effects. The crystal structure of the free-base form of bupivacaine has been determined directly from powder X-ray diffraction data using the Genetic Algorithm technique for structure solution, followed by Rietveld refinement. This work further emphasizes the scope and utility of ab initio structure solution directly from powder X-ray diffraction data for tackling structural problems within the biomedical field, leading to opportunities for the investigation of structure-property relationships. PMID- 14762906 TI - Chain length-dependent effects of alkylmaltosides on nasal absorption of enoxaparin. AB - The purpose of this study was to investigate whether the hydrophobic chain length of alkylmaltosides affects their efficacy as absorption promoters for nasally administered low-molecular-weight heparin and to study whether these agents enhance nasal absorption in a time-dependent manner without causing irreversible damage to the nasal epithelial membrane. For the nasal absorption studies, enoxaparin formulated with different alkylmaltosides was administered nasally to anesthetized rats and absorption of the drug was determined by measuring plasma anti-factor Xa activity. Reversibility studies were performed by administering enoxaparin at different time points after administration of alkylmaltosides. The AUC(0-360) for plasma anti-factor Xa-time curves increased with the increase in alkylmaltoside concentration in the formulations. Absolute and relative bioavailability of enoxaparin were increased by two-fold when the alkyl chain length of maltosides was increased from 8 to 14 carbons. Alkylmaltosides therefore increase nasal absorption of enoxaparin in a dose- and chain length dependent manner. Of the alkylmaltosides tested, tetradecylmaltoside is the most potent enhancer of nasal absorption of enoxaparin. Longer chain alkylmaltosides produce a more prolonged effect on nasal mucosa compared with those with shorter alkyl chain. PMID- 14762908 TI - Acid-base characteristics of bromophenol blue-citrate buffer systems in the amorphous state. AB - In this study, we have examined the acid-base characteristics of various citrate buffer systems alone and in the presence of the pH indicator dye, bromophenol blue, in aqueous solution, and after lyophilization to produce amorphous material. Fourier transform Raman and solid-state nuclear magnetic resonance spectroscopy have been used to monitor the ratio of ionized to un-ionized citric acid under various conditions, as a function of initial pH in the range of 2.65 4.28. Ultraviolet-visible spectrophotometry was used to probe the extent of proton transfer of bromophenol blue in the citrate buffer systems in solution and the amorphous state. Spectroscopic studies indicated greater ionization of citric acid and bromophenol blue in solution and the solid state with increasing initial solution pH, as expected. Fourier transform Raman measurements indicated the same ratio of ionized to un-ionized citrate species in solution, frozen solution, and the amorphous state. It is shown that the ratio of species at any particular initial pH is primarily determined by the amount of sodium ion present so as to maintain electroneutrality and not necessarily to the fact that pH and pK(a) remain unchanged during freezing and freeze drying. Indeed, for bromophenol blue, the relative ultraviolet-visible intensities for ionized and un-ionized species in the amorphous sample were different from those in solution indicating that the extent of protonation of bromophenol blue was significantly lower in the solid samples. It is concluded that under certain conditions there can be significant differences in the apparent hydrogen activity of molecules in amorphous systems. PMID- 14762907 TI - Optimizing storage stability of lyophilized recombinant human interleukin-11 with disaccharide/hydroxyethyl starch mixtures. AB - Optimal storage stability of a protein in a dry formulation depends on the storage temperature relative to the glass transition temperature (T(g)) of the dried formulation and the structure of the dried protein. We tested the hypothesis that optimizing both protein structure and T(g)--by freeze-drying recombinant human interleukin-11 (rhIL-11) with mixtures of disaccharides and hydroxyethyl starch (HES)--would result in increased storage stability compared with the protein lyophilized with either disaccharide or hydroxyethyl starch alone. The secondary structure of the protein in the dried solid was analyzed immediately after lyophilization and after storage at elevated temperatures by infrared spectroscopy. After rehydration, aggregation was monitored by size exclusion chromatography. Oxidation levels and cleavage products were quantified by reversed-phase chromatography. For the formulation with HES alone, which has a relatively high T(g), storage stability of rhIL-11 was poor, because HES failed to inhibit lyophilization-induced unfolding. The sugar formulations inhibited unfolding, and had intermediate T(g) values and storage stabilities. Addition of hydroxyethyl starch to sucrose or trehalose increased T(g) without affecting the capacity of the sugar to inhibit protein unfolding during lyophilization. Optimal storage stability of lyophilized rhIL-11 was achieved by using a mixture of disaccharide and polymeric carbohydrates. PMID- 14762909 TI - Incorporation of lipophilic drugs in sugar glasses by lyophilization using a mixture of water and tertiary butyl alcohol as solvent. AB - In this study, a new and robust method was evaluated to prepare physically stable solid dispersions. Trehalose, sucrose, and two inulins having different chain lengths were used as carrier. Diazepam, nifedipine, Delta(9) tetrahydrocannabinol, and cyclosporine A were used as model drugs. The sugar was dissolved in water and the drug in tertiary butyl alcohol (TBA). The two solutions were mixed in a 4/6 TBA/water volume ratio and subsequently freeze dried. Diazepam could be incorporated at drug loads up to 63% w/w. DSC measurements showed that, except in some sucrose dispersions, 97-100% of the diazepam was amorphous. In sucrose dispersions with high drug loads, about 10% of the diazepam had crystallised. After 60 days of exposure at 20 degrees C and 45% relative humidity (RH), diazepam remained fully amorphous in inulin dispersions, whereas in trehalose and sucrose crystallization of diazepam occurred. The excellent physical stability of inulin containing solid dispersions can be attributed to the high glass transition temperature (T(g)) of inulin. For the other drugs similar results were obtained. The residual amount of the low toxic TBA was only 0.1-0.5% w/w after freeze drying and exposure to 45% RH and 20 degrees C. Therefore, residual TBA will not cause any toxicity problems. This study provides a versatile technique, to produce solid dispersions. Inulin glasses are preferred because they provide an excellent physical stability of the incorporated amorphous lipophilic drugs. PMID- 14762910 TI - Stabilization and HPLC analysis of betamethasone sodium phosphate in plasma. AB - The analysis of corticosteroid prodrugs in pharmacokinetic (PK) studies poses the risk of overestimation of corticosteroid concentrations due to in vitro hydrolysis of prodrugs after sample collection. This study tests the effectiveness of enzyme inhibitors as stabilizers for betamethasone sodium phosphate (BSP) in pregnant sheep plasma samples collected during PK studies with betamethasone (BET) and provides simultaneous high-performance liquid chromatography analysis of BSP and BET. A rapid, sensitive, and specific ion paired reversed-phase high-performance liquid chromatography assay for simultaneous measurement of BET and BSP in plasma was developed. This assay was used for analyzing samples from an in vitro prodrug hydrolysis study. Enzyme inhibitors tested were sodium arsenate (Na(2)HAsO(4)) and ethylenediaminetetraacetic acid. The BSP was administered intramuscularly to three pregnant sheep to assess in vivo PK. Samples were split with part treated with Na(2)HAsO(4) and part left natural. In vitro hydrolysis of BSP in plasma to BET could be completely inhibited by Na(2)HAsO(4), but not by ethylenediaminetetraacetic acid. The PK study showed lower concentrations of BET in samples with Na(2)HAsO(4) compared with natural samples. This study demonstrates that artifacts in PK profiles of corticosteroids due to in vitro prodrug hydrolysis can be prevented by sample treatment with enzyme inhibitors. PMID- 14762911 TI - Partial characterization of different mixtures of solids by measuring the optical nonlinear response. AB - We report on the theoretical basis and first experimental results of a new method based on optical nonlinearity, for characterising crystallinity and polymorphism of pharmaceuticals in the solid state. Once the theoretical basis of optical nonlinearity of crystalline structures is established, a new and rapid method based on this physical theory can be developed to quantitatively determine polymorphism or crystallinity. An apparatus was set up to measure the second harmonic response of powdered samples when irradiated with a pulsed laser source. The response of quartz-glass, enalapril maleate forms I-II and enalapril maleate form II-PVP mixtures were measured and modeled. It was found that the quartz glass system showed high sensitivity to the presence of quartz and was well predicted by our theoretical model. The response of enalapril maleate polymorph mixtures was also sensitive to changes in the polymorph ratio. The theoretical predictions of the polymorph mixtures agreed quantitatively with the experimental results. The response of enalapril maleate form II-PVP mixtures agreed quantitatively with the physical model and showed extremely low noise and high sensitivity, giving very promising limits of detection (LOD) and quantification (LOQ) of 0.12 and 0.41%, respectively. This rapid, novel technique has potential for industrial monitoring of pharmaceutical manufacturing processes. PMID- 14762912 TI - Transport of octreotide and evaluation of mechanism of opening the paracellular tight junctions using superporous hydrogel polymers in Caco-2 cell monolayers. AB - The purpose of this study was to investigate the mechanism of opening of tight junctions in Caco-2 cell monolayers using superporous hydrogel (SPH) and SPH composite (SPHC) polymers as permeation enhancers for peptide drug delivery. Moreover, the transport of octreotide across Caco-2 cell monolayers was assessed by application of SPH and SPHC polymers on Caco-2 cell monolayers. In these experiments, N,N,N-trimethyl chitosan chloride with 60% quaternization (TMC60) was used as a positive control for opening of tight junctions. Transepithelial electrical resistance (TEER) studies showed that all three polymers (TMC60, SPH, and SPHC) were able to decrease TEER values to approximately 30% of the initial values, indicating the ability of these polymers to open the tight junctions. Recovery TEER studies showed that the effects of the polymers on Caco-2 cell monolayers were reversible, indicating viability of the cells after incubation with polymers. Both SPH and SPHC (compared with TMC60) were able to increase the paracellular transport of octreotide by their mechanical pressures on tight junctions. The mechanistic studies showed that junctional proteins, including actin, occludin, and claudin-1, were influenced by application of SPH and SPHC polymers to the Caco-2 cell monolayers. SPH and SPHC induced clear changes in the staining pattern of all three proteins compared with the control, indicating that the expression of these proteins in the tight junctions was increased, most likely due to the mechanical pressure of the polymers on the junctional proteins. PMID- 14762913 TI - The influence of relative humidity on the cohesion properties of micronized drugs used in inhalation therapy. AB - The influence of relative humidity (RH) on the cohesion properties of three drugs: salbutamol sulphate (SS), triamcinolone acetonide (TAA), and disodium cromoglycate (DSCG) was investigated using the atomic force microscope (AFM) colloidal probe technique. Micronized drug particles were mounted in heat sensitive epoxy resin for immobilization. Multiple AFM force-distance curves were conducted between each drug probe and the immobilized drug particulates at 15, 45, and 75% RH using Force-Volume imaging. Clear variations in the cohesion profile with respect to RH were observed for all three micronized drugs. The calculated force and energy of cohesion to separate either micronized SS or DSCG increased as humidity was raised from 15 to 75% RH, suggesting capillary forces become a dominating factor at elevated RH. In comparison, the separation force and energy for micronized TAA particles decreased with increased RH. This behavior may be attributed to long-range attractive electrostatic interactions, which were observed in the approach cycle of the AFM force-distance curves. These observations correlated well with previous aerosolization studies of the three micronized drugs. PMID- 14762914 TI - Mechanistic studies of branched-chain alkanols as skin permeation enhancers. AB - As part of a long-term effort to understand the structure/function relationship between chemical permeation enhancers and skin permeation enhancement, the present study examined the influence of hydrocarbon chain branching on the effectiveness of skin permeation enhancers of the type that possesses a polar group (e.g., the hydroxyl group) attached to a hydrocarbon chain(s). The effects of x-hexanol, x-heptanol, x-octanol, and x-nonanol (where x is the position of the hydroxyl group ranging from 1 up to 5) on the transport of a probe permeant, corticosterone, 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, across the lipoidal pathway of stratum corneum (SC). The isoenhancement concentrations of 2-alkanol, 3-alkanol, 4-alkanol, and 5-alkanol to induce E = 10 were approximately 1.9-, 2.6-, 3.1-, and 3.9-fold higher, respectively, than those of the 1-alkanols of the same molecular formula. This suggested that the branched-chain alkanols have lower enhancer potency than the 1-alkanols of the same molecular formula; the potency decreases as the hydroxyl group moves from the end of the chain towards the center of the enhancer alkyl chain. To further investigate the mechanism(s) of action of the branched-chain alkanols as skin permeation enhancers, the equilibrium uptake of the enhancers into the hairless mouse skin stratum corneum (HMS SC) from aqueous enhancer solutions of E = 10 was determined. The data from these experiments provided a direct measure of the "intrinsic" potency of the enhancer. In the same experiments, the equilibrium partitioning (distribution) of a surrogate permeant, estradiol (E2beta), into the HMS SC was also determined and compared to the partitioning from PBS (no enhancer present). The uptake amounts (micromole/mg SC) for 1-alkanols into the intercellular lipids of the SC were found to be essentially the same at their isoenhancement concentrations. However, at their isoenhancement concentrations, the uptake amounts of the branched-chain alkanols into the intercellular lipids of HMS SC were higher than those of the 1-alkanols. These results support the view that: (1) the intrinsic potencies of the 1-alkanols are essentially the same and independent of their 1-alkyl chain length at their isoenhancement concentrations, (2) the intrinsic potencies of the branched-chain alkanols are lower than those of the normal alkanols, and (3) branching of the alkyl chain reduces the ability of the enhancer to effect lipid fluidization in the SC lipid lamellae at the target site(s). The enhancement effects of the branched-chain alkanols and the 1-alkanols at their isoenhancement concentrations upon E2beta partitioning into the SC intercellular lipids were found to be approximately the same and in the range of five- to eight-fold enhancement. The constancy of this enhancement for E2beta partitioning suggests that the mechanism of enhancement action for the branched-chain alkanols and the 1-alkanols are the same. Additionally, a good correlation of the intercellular lipid/PBS partition coefficients of both the branched-chain alkanols and the 1-alkanols with the n octanol/PBS partition coefficients was found. This supports the view that the chemical microenvironment of the polar head group and the alkyl group of the studied enhancers at the site of skin permeation enhancer action in the SC lipid lamellae can be represented by water-saturated n-octanol for both the branched chain alkanols and the 1-alkanols. PMID- 14762915 TI - The characterization of fluidization behavior using a novel multichamber microscale fluid bed. AB - In the preformulation stage, there is a special need to determine the process behavior of materials with smaller amounts of samples. The purpose of this study was to assemble a novel automated multichamber microscale fluid bed module with a process air control unit for the characterization of fluidization behavior in variable conditions. The results were evaluated on the basis of two common computational methods, the minimum fluidization velocity, and the Geldart classification. The materials studied were different particle sizes of glass beads, microcrystalline cellulose, and silicified microcrystalline cellulose. During processing, the different characteristic fluidization phases (e.g., plugging, bubbling, slugging, and turbulent fluidization) of the materials were observed by the pressure difference over the bed. When the moisture content of the process air was increased, the amount of free charge carriers increased and the fine glass beads fluidized on the limited range of velocity. The silicification was demonstrated to improve the fluidization behavior with two different particle sizes of cellulose powders. Due to the interparticle (e.g., electrostatic) forces of the fine solids, the utilization of the computational predictions was restricted. The presented setup is a novel approach for studying process behavior with only a few grams of materials. PMID- 14762916 TI - Physical stability of salmon calcitonin spray-dried powders for inhalation. AB - The effects of excipient crystallinity and water content on the physical stability of salmon calcitonin (sCT) in a spray-dried powder for inhalation have been investigated. sCT was dissolved in water with and without mannitol and then spray dried using a Buchi 190 spray dryer. The spray dried powders were stored for 5 days at 0, 29, 51, 58, 69, and 84% relative humidity at ambient temperature. The crystalline content, water content, secondary structure, and aggregation rates were determined for each powder immediately following spray drying and after storage at various relative humidities. In addition, the water sorption isotherms and reactivity to water vapor were determined using DVS and isothermal calorimetry, respectively. No sCT aggregation occurred during the spray drying process. Crystallinity depended on the amount of mannitol in the formulation. Powders containing up to 50% mannitol were fully amorphous, and those containing 70 and 90% mannitol contained some crystalline polyol. The powders remained aggregate free for over 2 years when stored below the critical RH (e.g., <20% for the powder containing 30% mannitol). Above this RH, sCT aggregation increased as a function of time. The amount of aggregate observed correlates with the amount of intermolecular beta-sheet formed, determined by FTIR. The sCT aggregation rate in powders containing 70% mannitol was significantly lower than that in powders containing 30% mannitol at all RH tested, presumably because of a higher ratio of amorphous mannitol to sCT, which inhibits the formation of beta-sheet structure. Moisture-induced crystallization of mannitol was observed in all powders stored at RH >50%. The moisture induced thermal activity trace (MITAT) offers a useful description on the physical stability of the spray dried powders. In conclusion, spray drying sCT and sCT/mannitol mixtures yields dry powders that contain physically intact peptide. In addition, sCT aggregation and mannitol crystallization in spray dried powders can be prevented during long-term storage if stored in low humidity environments, which can be easily assessed by MITAT. PMID- 14762919 TI - Evaluations of bioactivity and mechanical properties of poly (epsilon caprolactone)/silica nanocomposite following heat treatment. AB - A composite material consisting of poly(epsilon-caprolactone) (PCL) and silica was prepared and evaluated as a bioactive bone substitute. The composite was synthesized by the co-condensation of tetraethyl orthosilicate and PCL and end capped with triethoxysilane (Si-PCL). The as-prepared specimens were subjected to an initial heat treatment of 2 days at 60 degrees C, followed by further heat treatments at 100 degrees C, 150 degrees C, and 200 degrees C for 24 h. The tensile mechanical properties of the heat-treated specimens were determined, and additional specimens were exposed to a simulated body fluid (SBF) for different periods of time. The SBF exposure led to the deposition of a layer of apatite crystals on the surface of the composites. It was found that increasing the second heat-treatment temperature produced an increase in tensile strength and Young's modulus of the composite but a decrease in the initial rate of apatite formation. These phenomena are explained in terms of the condensation reaction that takes place between the silanol groups in the silica and Si-PCL as the heat treatment temperature is increased. PMID- 14762920 TI - Evaluation of biocompatibility of polypyrrole in vitro and in vivo. AB - In this study, the biocompatibility of the electrically conductive polymer polypyrrole (PPy) with nerve tissue was evaluated in vitro and in vivo. The extraction solution of PPy powder, which was synthesized chemically, was tested for acute toxicity, subacute toxicity, pyretogen, quantitative measure of cell viability, hemolysis, allergen, and micronuclei. The PPy membrane was synthesized electrochemically on the indium tin oxide conductive borosilicate glass. The dorsal root ganglia from 1-3-day-old Sprague-Dawley rats were cultured above PPy membrane and observed by light or scanning electron microscopy. The PPy-silicone tube (PPy membrane on the inner surface of the silicone tube) also synthesized electrochemically was used to bridge across 10-mm sciatic nerve gap in rats. Twenty-four weeks after the operation to rats, the regenerated tissues were observed by electrophysiological and histological techniques. PPy extraction solution showed no evidence of acute and subacute toxicity, pyretogen, hemolysis, allergen, and mutagenesis, and the Schwann cells from the PPy extraction solution group showed better survival rate and proliferation rate as compared with the saline solution control group. The migration of the Schwann cells and the neurite extension from dorsal root ganglia on the surface of PPy membrane-coated glass was better than those of bare glass. There was only lightly inflammation during 6 months of the postoperation, when the PPy-silicone tube bridged across the gap of the transected sciatic nerve. The regeneration of nerve tissue in the PPy silicone tube was slightly better than that in the plain silicone tube by means of electrophysiological and histological examination. The results of this study indicate that PPy has a good biocompatibility with rat peripheral nerve tissue and that PPy might be a candidate material for bridging the peripheral nerve gap. PMID- 14762921 TI - Ongoing foreign body reaction to subcutaneous implanted (heparin) modified Dacron in rats. AB - Dacron-containing heart valve repair devices trigger chronic inflammation characterized by the presence of activated macrophages, foreign body giant cells, and capsule formation. Upon blood contact, proinflammatory proteins adsorb to the material and provide a substrate for monocyte binding and differentiation. Various heparin-coated polymers have been shown to reduce adsorption of proinflammatory proteins in vitro and in vivo. In this study, the effect of knitted, heparin-coated Dacron on the foreign body reaction was tested subcutaneously in rats. We hypothesized that the anti-inflammatory effect of heparin would reduce monocyte recruitment and differentiation and therefore limit the inflammatory reaction. An ongoing foreign body reaction, characterized by the presence of foreign body giant cells and high vascularization, was observed in uncoated as well as (heparin-)coated Dacron at up to 180 days of implantation. Also, a thin capsule was formed around each material up to this time. In conclusion, although heparin coatings might have an effect on the acute inflammatory response, we were not able to show a difference between heparin coated and uncoated Dacron after 180 days' implantation in rats. Further research needs to be conducted to assess the difference in proinflammatory protein adsorption between the tested materials and the effect this has on the long-term foreign body reaction. PMID- 14762922 TI - Effect of biomaterial properties on bone healing in a rabbit tooth extraction socket model. AB - In this work we sought to understand the effect of biomaterial properties upon healing bone tissue. We hypothesized that a hydrophilic polymer gel implanted into a bone tissue defect would impede the healing process owing to the biomaterial's prevention of protein adsorption and thus cell adhesion. To test this hypothesis, healing bone was investigated within a rabbit incisor extraction socket, a subcritical size bone defect that resists significant soft tissue invasion by virtue of its conformity. After removal of the incisor teeth, one tooth socket was left as an empty control, one was filled with crosslinked polymer networks formed from the hydrophobic polymer poly(propylene fumarate) (PPF), and one was filled with a hydrogel formed from the hydrophilic oligomer oligo(poly(ethylene glycol) fumarate) (OPF). At five different times (4 days as well as 1, 2, 4, and 8 weeks), jaw bone specimens containing the tooth sockets were removed. We analyzed bone healing by histomorphometrical analysis of hematoxylin and eosin stained sections as well as immunohistochemically stained sections. The proposed hypothesis, that a hydrophilic material would hinder bone healing, was supported by the histomorphometrical results. In addition, the immunohistochemical results reflect molecular signaling indicative of the early invasion of platelets, the vascularization of wound-healing tissue, the differentiation of migrating progenitor cells, and the formation and remodeling of bone tissue. Finally, the results emphasize the need to consider biomaterial properties and their differing effects upon endogenous growth factors, and thus bone healing, during the development of tissue engineering devices. PMID- 14762923 TI - Responsive and recognitive hydrogels using star polymers. AB - Polymeric networks that have inherent capabilities to recognize different molecules and chemical changes in their environment are the next generation of materials that will aid in the diagnosis and treatment of diseases. We have prepared new networks based on star polymers that were designed to be responsive and recognitive. Using molecular imprinting with D-glucose and crosslinking with poly(ethylene glycol) dimethacrylate with an ethylene glycol chain of nominal molecular weight 600, we prepared star polymer networks, which exhibited over 300% more uptake for D-glucose compared to D-fructose. Using copolymerization with methacrylic acid, we prepared star polymer networks with pH-sensitivity, which showed a sharp transition in swelling around a pH of 4.5. PMID- 14762924 TI - Anti-inflammatory effects of a titanium-peroxy gel: role of oxygen metabolites and apoptosis. AB - Polymorphonuclear neutrophils (PMN) are among the first inflammatory cells to arrive at an implant interface, where they encounter with the foreign material and may produce reactive oxygen species (ROS). During the interaction between titanium and ROS, titanium-peroxy (Ti-peroxy) compounds may be formed. We used a Ti-peroxy gel, made from titanium and hydrogen peroxide, to study the effects of Ti-peroxy compounds on PMN. In the absence of serum, the Ti-peroxy gel decreased the oxidative response of PMN to yeast and PMA and reduced PMN apoptosis without inducing necrosis. These effects could not be ascribed to the release of hydrogen peroxide from the Ti-peroxy gel, because a steady-state hydrogen peroxide producing system failed to mimic the effects of the gel. The effects were similarly unaffected when PMN were preincubated with beta(2)-integrin antibodies, questioning the involvement of adhesion molecules. Nevertheless, when a filter was used to separate the Ti-peroxy gel from the cells, the gel effect on PMN life span was abolished, pointing to a contact-dependent mechanism. In the presence of serum, the Ti-peroxy gel had no effect on the PMN oxidative response and life span, but appeared rather inert. In summary, this study demonstrates that the Ti peroxy gel has potentially anti-inflammatory properties through a combined peroxide and physical contact effect, supporting the notion that interactions between titanium and inflammatory cells are responsible for the good performance of titanium in vivo. PMID- 14762925 TI - RGD-containing peptide GCRGYGRGDSPG reduces enhancement of osteoblast differentiation by poly(L-lysine)-graft-poly(ethylene glycol)-coated titanium surfaces. AB - Osteoblasts exhibit a more differentiated morphology on surfaces with rough microtopographies. Surface effects are often mediated through integrins that bind the RGD motif in cell attachment proteins. Here, we tested the hypothesis that modulating access to RGD binding sites can modify the response of osteoblasts to surface microtopography. MG63 immature osteoblast-like cells were cultured on smooth (Ti sputter-coated Si wafers) and rough (grit blasted/acid etched) Ti surfaces that were modified with adsorbed monomolecular layers of a comb-like graft copolymer, poly-(L-lysine)-g-poly(ethylene glycol) (PLL-g-PEG), to limit nonspecific protein adsorption. PLL-g-PEG coatings were functionalized with varying amounts of an integrin-receptor-binding RGD peptide GCRGYGRGDSPG (PLL-g PEG/PEG-RGD) or a nonbinding RDG control sequence GCRGYGRDGSPG (PLL-g-PEG/PEG RDG). Response to PLL-g-PEG alone was compared with response to surfaces on which 2-18% of the polymer sidechains were functionalized with the RGD peptide or the RDG peptide. To examine RGD dose-response, peptide surface concentration was varied between 0 and 6.4 pmol/cm(2). In addition, cells were cultured on uncoated Ti or Ti coated with PLL-g-PEG or PLL-g-PEG/PEG-RGD at an RGD surface concentration of 0.7 pmol/cm(2), and free RGDS was added to the media to block integrin binding. Analyses were performed 24 h after cultures had achieved confluence on the tissue culture plastic surface. Cell number was reduced on smooth Ti compared to plastic or glass and further decreased on surfaces coated with PLL-g-PEG or PLL-g-PEG/PEG-RDG, but was restored to control levels when PLL g-PEG/PEG-RGD was present. Alkaline phosphatase specific activity and osteocalcin levels were increased on PLL-g-PEG alone or PLL-g-PEG/PEG-RDG, but PLL-g-PEG/PEG RGD reduced the parameters to control levels. On rough Ti surfaces, cell number was reduced to a greater extent than on smooth Ti. PLL-g-PEG coatings reduced alkaline phosphatase and increased osteocalcin in a manner that was synergistic with surface roughness. The RDG peptide did not alter the PLL-g-PEG effect but the RGD peptide restored these markers to their control levels. PLL-g-PEG coatings also increased TGF-beta1 and PGE(2) in conditioned media of cells cultured on smooth or rough Ti; there was a 20x increase on rough Ti coated with PLL-g-PEG. PLL-g-PEG effects were inhibited dose dependently by addition of the RGD peptide to the surface. Free RGDS did not decrease the effect elicited by PLL g-PEG surfaces. These unexpected results suggest that PLL-g-PEG may have osteogenic properties, perhaps correlated with effects that alter cell attachment and spreading, and promote a more differentiated morphology. PMID- 14762926 TI - Macroporous poly(epsilon-caprolactone) with antimicrobial activity obtained by iodine polymerization. AB - The most serious problem usually encountered in the field of implanted biomedical devices is infectious morbidity as a primary source of mortality. In this work, the synthesis and characterization of a macroporous iodine-based sanitizer (iodophor), poly(caprolactone)-iodine (PCL-I(2)), are presented. Characterization methods include nuclear magnetic resonance spectroscopy, gel permeation chromatography, nitrogen adsorption-desorption, and scanning electron microscopy. The in vitro cytotoxicity to CHO cells based on cell viability with Chinese hamster ovary cells (CHO) and antimicrobial activities against Escherichia coli and Staphylococcus aureus were examined. The obtained macropore PCL-I(2) structures had a rather narrow size distribution. The PCL-I(2) iodophor was noncytotoxic to Chinese hamster ovary cells. The antimicrobial activities of the PCL-I(2) were assessed against E. coli and S. aureus. The tested PCL-I(2) showed better antimicrobial activity against E. coli than against S. aureus. PMID- 14762927 TI - Vitronectin adsorption on surfaces visualized by tapping mode atomic force microscopy. AB - Vitronectin (Vn) is an abundant glycoprotein present in plasma and the extracellular matrix of most tissue and is an important medium required for bone cells to attach and spread on biomaterial surfaces. Hence, Vn adsorption is an initial and key step in implantation. In this study, adsorption of Vn molecules on mica and oxidized titanium substrates in Milli-Q water (pH 5.6) and in simulated body fluid (SBF, pH 7.4) has been investigated using tapping mode atomic force microscopy. Conformation of the adsorbed proteins was determined from the images. The effect of adsorption time was investigated on mica surfaces. The protein first attaches to the surface, then immediately spreads out with different lateral dimensions of adsorbed protein and becomes aggregated. After 15 min of adsorption, globular Vn molecules form clusters of aggregates in extended chains on the mica surface. Globular Vn molecules appear flatter (i.e., oblate ellipsoids) and coalesce on the ridges of the uneven oxidized titanium surface. From cross-sectional analyses of the Vn images, it was found that the contact areas of Vn molecules associated with Ti surfaces are larger than those with mica surfaces. This suggests that the different surface properties of substrates contribute to this different conformation of adsorbed Vn molecules. The larger contact areas of Vn associated with Ti substrates indicates higher affinity for the biomaterial oxidized titanium surface than for mica. In SBF, Vn interacted weakly with the mica surfaces and Vn molecules were easily desorbed during the imaging process. On oxidized titanium substrates, after 3 h of incubation time in Vn-SBF solution, images can be obtained showing adsorbed Vn aggregates on the oxidized titanium surface. In this case, the protein colloids are strongly adherent, with increased lateral dimensions compared with the adsorption on mica. PMID- 14762928 TI - Polymeric sustained local drug delivery system for the prevention of vascular intimal hyperplasia. AB - Anastomotic intimal hyperplasia (IH) is a major cause of both autologous vein and synthetic vascular graft failure. We have previously published data suggesting that cyclosporin may reduce the development of IH in a canine model. However, systemic administration of cyclosporin could create serious adverse effects. Therefore, it is our long-term goal to test the hypothesis that the controlled local release of cyclosporin from a polymeric vascular wrap will prevent the development of IH. To test this hypothesis, we developed a controlled release vascular wrap (sheet/ring) using a poly(ethylene glycol) (PEG) hydrogel. Sterilization of the polymers was performed using the ethylene oxide and hydrogen peroxide sterilization methods. It was found that except for one combination (8000 molecular weight and 1:1 crosslinking ratio), the differences in the swelling ratios for the sterilized and unsterilized hydrogels were not statistically significant. Release studies from unsterilized and ethylene oxide sterilized PEG hydrogels were conducted. It was found that release lasted for approximately 50 h for sterilized as well as unsterilized PEG hydrogels. Acute animal studies, to test the deployment of both the polymeric sheets and rings to the adventitial surface of native arteries and veins, were completed successfully. PMID- 14762929 TI - Small changes in the polymer structure influence the adsorption behavior of fibrinogen on polymer surfaces: validation of a new rapid screening technique. AB - Numerous studies conclude that the selective adsorption of plasma proteins on materials contacting blood or tissue affects all subsequent interactions related to the biocompatibility of artificial surfaces. However, there are only a few studies available, which clearly demonstrate that there is a correlation between surface chemistry and selective protein adsorption. Detailed knowledge of such correlations would facilitate the design of biocompatible materials. In this study, a rapid, fluorescence-based, screening technique using a 384-well format for polymer-protein interactions was developed. The screening assay was used to measure the adsorption of human fibrinogen on 46 test polymers (44 polyarylates selected from a combinatorial library of tyrosine-derived polyarylates, and two lactide-based polymers). In this library of polyarylates, structural changes are generated by variations in either the polymer backbone or the polymer pendent chain. Although no overall trend between polymer hydrophobicity and fibrinogen adsorption could be identified using the entire set of test polymers (R(2) = 0.43), fibrinogen adsorption was clearly correlated with variations in the pendent chain structure. Thus, when the test polymers were grouped by backbone composition, increased hydrophobicity of the pendent chain was significantly correlated with reduced fibrinogen adsorption. The following R(2) coefficients within the polymer backbone groups were determined: 0.87 (diglycolates); 0.98 (glutarates); 0.73 (adipates); 0.87 (suberates); 0.67 (3-methyl-adipates). Our results demonstrate that it is possible to screen for protein-material interactions in a cost-effective fashion using a miniaturized immunofluorescence technique. Further, we demonstrate that small changes in chemical composition can significantly influence the adsorption of human fibrinogen on polymer surfaces. The lactide-based polymers were among those polymers exhibiting the highest tendency to adsorb fibrinogen. This information may be useful when polymers have to be selected for specific biomaterial applications. PMID- 14762930 TI - Systematic variation in osteoblast adhesion and phenotype with substratum surface characteristics. AB - Time-varying interactions of human fetal osteoblastic cells (hFOB 1.19) with materials of diverse chemical composition and surface energy, including biodegradable lactide/glycolide-based polymers, were assessed using a combination of assays sensitive to different phases of cell-substratum compatibility. Short term (minutes to hours) cell-attachment-rate assays were used to measure the earliest stages of cell-surface interactions leading to adhesion. Proliferation rate assays quantifying viability of attached cells were applied as a measure of medium-term (hours to days) cytocompatibility. Both attachment- and proliferation rate assays were found to strongly correlate with material surface energy, with the exception of a reproducible and significant adhesion preference for fully water-wettable quartz over glass. No such adhesion/proliferation preference was observed for hydrophobized counterparts, and attachment to water-wettable glass was significantly less than that to control tissue culture polystyrene. These results suggest that the amorphous SiO(x) surface was inhibitory to hFOB 1.19 growth whereas putatively crystalline quartz stimulated bioadhesion. Alkaline phosphatase activity was evaluated as a marker for long-term (days) differentiation of hFOB 1.19 cells and did not strongly correlate with surface energy or, in the case of biodegradable polymers, chemical composition. PMID- 14762931 TI - In vivo cancellous bone remodeling on a strontium-containing hydroxyapatite (sr HA) bioactive cement. AB - The purpose of this study was to investigate the in vivo bone response to the strontium-containing hydroxyapatite (Sr-HA) bioactive bone cement injected into the cancellous bone. Sr-HA cement was injected into the iliac crest of rabbits for 1, 3, and 6 months. Active bone formation and remodeling were observed after 1 month. Newly formed bone was observed to grow onto the bone cement after 3 months. Thick osteoid layer with osteoblasts formed along the bone and guided over the bone cement surface reflected the stimulating effect of Sr-HA. From scanning electron microscopy (SEM) and energy-dispersive X-ray (EDX) analysis, high calcium and phosphorus levels were detected at the interface with a thick layer of 70 microm in width, and fusion of Sr-HA with the bone was observed. Blood vessels were found developing in remodeling sites. The affinity of bone on Sr-HA cement was increased from 73.55 +/- 3.50% after 3 months up to 85.15 +/- 2.74% after 6 months (p < 0.01). In contrast to Sr-HA cement, poly(methyl methacrylate) (PMMA) bone cement was neither osteoconductive nor bioresorbable. Results show that the Sr-HA cement is biocompatible and osteoconductive, which is suitable for use in treating osteoporotic vertebral fractures. PMID- 14762932 TI - Dissolution control and cellular responses of calcium phosphate coatings on zirconia porous scaffold. AB - Different types of calcium phosphates [hydroxyapatite (HA), fluorapatite (FA), tricalcium phosphate (TCP), and their composites (HA + FA, HA + TCP)] were coated on a zirconia (ZrO(2)) porous scaffold using a powder slurry method. The ZrO(2) porous scaffold was intended for a load-bearing implant, and the apatite layers were coated to improve osteoconductivity. The insertion of an FA intermediate layer between the coating layer and ZrO(2) scaffold effectively suppressed the reaction between the calcium phosphate and ZrO(2) and maintained the coating layer at the initial powder composition. The obtained coating layer, of a thickness of approximately 30 microm, was relatively microporous and firmly adherent to the ZrO(2) scaffold. Dissolution tests in physiological solution showed typical differences depending on the coating layers, with the dissolution rate increasing in the order TCP > HA + TCP > HA > HA + FA > FA. This result suggests the functional coating of the calcium phosphates in view of tailoring the solubility. Osteoblast-like cells, MG63 and HOS, responded similarly in terms of cell growth, morphology, and proliferation rate regardless of the coating types, indicating favorable and comparable cell viability. However, the alkaline phosphatase (ALP) activity of the cells on the pure HA and HA composite coatings (HA + FA and HA + TCP) expressed at higher levels compared to those on pure FA and pure TCP coatings for both MG63 and HOS cells, suggesting a selective cell activity depending on the coating types. All the calcium phosphate-coated-ZrO(2) scaffolds showed higher ALP levels compared to pure ZrO(2) scaffold. PMID- 14762933 TI - Effect of cyclic mechanical stretch and titanium particles on prostaglandin E2 production by human macrophages in vitro. AB - Early implant instability has been proposed as a critical factor in the onset and progression of aseptic loosening and periprosthetic osteolysis in total joint arthroplasties. Previous in vitro studies have reported that macrophages stimulated with cyclic mechanical strain release inflammatory mediators. Little is known, however, about the response of these cells to mechanical strain with particles, which is often a component of the physical environment of the cell. We therefore studied the production of prostaglandin E(2) (PGE(2)), an important mediator in aseptic loosening and periprosthetic osteolysis in total joint arthroplasties, for human macrophages treated with mechanical stretch alone, titanium particles alone, and mechanical stretch and particles combined. A combination of mechanical stretch and titanium particles resulted in a statistically synergistic elevation of levels of PGE(2) compared with the levels found with either stretch or particles alone. Exposure of human macrophages to mechanical stretch with particles upregulated the expression of cyclooxygenase (COX)-2 mRNA but not COX-1 mRNA, this expression resulting in a 97-fold increase in PGE(2) production compared to the nonstimulated cells. The current study is the first to investigate the effects of mechanical stretch with particles on cultured macrophages and include an investigation of the time course of PGE(2) production and COX-2 mRNA expression. Our results suggest that, while mechanical strain may be one of the primary factors responsible for macrophage activation and periprosthetic osteolysis, mechanical strain with particles load may contribute significantly to the osteolytic potential of macrophages in vitro. The synergistic effect observed between mechanical stretch and particles could accelerate implant loosening and implies that reduction in either cyclic mechanical strain or wear debris load would lead to a reduction of osteolysis. PMID- 14762934 TI - Transforming growth factor-beta1 accelerates bone bonding to a blended calcium phosphate ceramic coating: a dose-response study. AB - In vivo strain measurements can facilitate the study of the bone remodeling response to loading and load changes. Calcium phosphate ceramic (CPC) coatings have been used to attach strain gauges to bone for extended periods of time, but require up to 12 weeks for adequate CPC-to-bone bonding. Transforming growth factor beta 1 (TGF-beta1), an osteoinductive growth factor, was used as a surface enhancement to accelerate bone growth and bonding to CPC particles. The aim of this study was to find an optimal dosage of TGF-beta1 to accelerate the attachment process. CPC-coated strain gauges were enhanced with doses of 0.5, 1.0, or 2.0 microg of TGF-beta1 per gauge. Gauges were placed on the femora of dogs, which were exercised daily and fed ad libitum. After 3, 6, and 12 weeks, gauge attachment was quantitatively assessed using mechanical testing and histomorphometry. Gauge attachment was also qualitatively assessed using back scatter electron microscopy. Agreement of the mechanical test results with both the back scatter electron microscopy images and histomorphometry results showed that the 1.0 microg per gauge dose of TGF-beta1 is an optimal dose to accelerate bone formation and attachment to CPC-coated strain gauges. PMID- 14762935 TI - Scaled interfacial activity of proteins at the liquid-vapor interface. AB - A principal conclusion drawn from observations of time- and concentration dependent liquid-vapor (LV) interfacial tension gamma(lv) of a diverse selection of proteins ranging from albumin to ubiquitin spanning nearly three decades in molecular weight (MW) is that concentration scaling substantially alters perception of protein interfacial activity as measured by reduction in gamma(lv). Proteins appear more similar than dissimilar on a weight/volume basis, whereas molarity scaling reveals a "Traube-rule" ordering by MW, suggesting that adsorption is substantially driven by solution concentration rather than diversity in protein amphilicity. Scaling as a ratio-to-physiological concentration demonstrates that certain proteins exhibit the full possible range of interfacial activity at and well-below physiological concentration, whereas others are only weakly surface active within this range, requiring substantially higher solution concentration to achieve reduction in gamma(lv). Important among this latter category of proteins are the blood factors XII and XIIa, assumed by the classical biochemical mechanism of plasma coagulation to adsorb to procoagulant surfaces, even in the presence of overwhelming concentrations of other blood constituents such as albumin and immunoglobulin that are shown by this work to be among the class of highly surface-active proteins at physiologic concentration. A comparison of pendant drop and Wilhelmy balance tensiometry as tools for assessing protein interfacial activity shows that measurement conditions employed in the typical Wilhelmy plate approach fails to achieve the steady-state adsorption condition that is accessible to pendant drop tensiometry. PMID- 14762936 TI - AFM observation of collapse and expansion of phosphoric acid-demineralized dentin. AB - The objective of this study is to provide additional data regarding morphological changes that occur to dentin matrices following demineralization with etchants. Our understanding of the mechanism of diffusion of comonomers into the demineralized substrate is very limited. It has been hypothesized that certain water-soluble polyelectrolytes (acidic proteins) and neutral proteins in dentin can influence the collapse of demineralized dentin when it is air dried. Some of these solubilized substances are thought to aggregate by the action of Ca cations, which become dissolved during H(3)PO(4) etching, ultimately resulting in some degree of collapse. In the current study, dentin surfaces were examined by atomic force microscopy (AFM) before and after treatment by 10% H(3)PO(4)containing increasing concentrations of CaHPO(4). Reversal of matrix collapse by aqueous 30% 2-hydroxyethyl methacrylate (HEMA) was evaluated by AMF for 60 min. The results demonstrate two forms of matrix collapse; we speculate that one form is induced by acidic noncollagenous polyelectrolytes and the other by neutral peptides. Our data indicate that further evaluation of the influence of endogenous noncollagenous proteins must be studied to understand the mechanism of the collapse and reexpansion dynamics of demineralized dentin networks. PMID- 14762937 TI - Improved wet bonding of methyl methacrylate-tri-n-butylborane resin to dentin etched with ten percent phosphoric acid in the presence of ferric ions. AB - The objective of this study was to determine the influence of dissolved dentinal substances in demineralized dentin on the hybridization of resin for bonding to dentin. It was hypothesized that these substances, including polyelectrolytes, significantly change the substrates, which could then be assessed by the addition of Na(+), Ca(2+), or Fe(3+) in 10% phosphoric acid. Bovine dentin specimens were etched for 10 s with a solution of 10% phosphoric acid (control) or of 22.0 mM dissolved sodium chloride (10P-Na), calcium chloride (10P-Ca), or ferric chloride (10P-Fe). The specimens were then rinsed, blot-dried, and primed three times with 5% 4-methacryloyloxyethyl trimellitate anhydride in acetone for 60 s. Methyl methacrylate-tri-n-butylborane resin was then applied. The tensile bond strength of each of the dumbbell-shaped specimens was then measured. The fractured surfaces and modified cross-sections were examined by scanning electron microscopy. The cross-sections were soaked in 6N HCl for 10 s and then in 1% sodium hypochlorite for 30 min to determine the resin content in the hybridized specimens. Shrinkage of the demineralized dentins upon drying was assessed by atomic force microscopy. The tensile bond strengths were 10.8 +/- 4.5 (control), 15.0 +/- 7.0 (10P-Na), 19.3 +/- 5.5 (10P-Ca), and 27.8 +/- 8.1 (10P-Fe) MPa. The atomic force microscopy studies showed that Fe(3+) minimized the shrinkage by drying for 10 s but Ca(2+) and Na(+) did not decrease the shrinkage the same as the control. The results support the hypothesis that the monomer permeability of wet demineralized dentin is effectively improved by dissolving ferric ions in the phosphoric acid, resulting in a greater bond strength and higher resin content in the hybridized dentin. The dissolved dentinal substances, including the polyelectrolytes, had a significant influence on the characteristics of the demineralized dentin, changing the degree of hybridization and bonding. PMID- 14762938 TI - Effects of end boundary conditions and specimen geometry on the viscoelastic properties of cancellous bone measured by dynamic mechanical analysis. AB - The viscoelastic properties of cancellous bone can be measured nondestructively in compression testing using a dynamic mechanical analyzer. In this study, we examined the effects of end boundary conditions and specimen geometry on the viscoelastic properties of cancellous bone measured by dynamic mechanical analysis. During dynamic compression testing, the cancellous bone specimens may be mechanically fixed (e.g., glued) to the loading platens or they may be free to expand across the platen surface. When specimens of cancellous bone were tested between platens with gluing, the dependence of loss tangent on frequency was not consistent with previously observed strain-rate-dependent mechanical behavior of cancellous bone. When long specimens of cancellous bone (length = 10 mm, diameter = 8 mm) were tested without gluing, the relationship between loss tangent and frequency depended on the level of load applied. For short specimens (length = 5 mm, diameter = 8 mm) tested without gluing, however, the frequency dependence of loss tangent agreed with existing data reported for the strain-rate-dependent behavior of cancellous bone and also with the frequency dependence of cortical bone viscoelasticity. Therefore, we recommend that short cancellous bone cylinders with a length of 5 mm and a diameter of 8 mm should be used without gluing in the dynamic mechanical analysis of cancellous bone. This is consistent with the American Society for Testing and Materials testing recommendations for plastics, but different from current practice for unimodal mechanical testing of cancellous bone. PMID- 14762939 TI - Biocompatibility of chemoenzymatically derived dextran-acrylate hydrogels. AB - The biocompatibility of chemoenzymatically generated dextran-acrylate hydrogels has been evaluated in vitro, using human foreskin fibroblasts, and in vivo, by subcutaneous and intramuscular implantation in Wistar rats for up to 40 days. In vitro tests show that hydrogel extracts only minimally reduced (<10%) the mitochondrial metabolic activity of fibroblasts. Direct contact of the hydrogels with cells induced a cellular proliferation inhibition index (CPII) of 50-80%, compared with a control, whereas through indirect contact, the CPII values were <16%, suggesting that the high CPII values achieved in the direct assay test were likely due to mechanical stress or limitations in oxygen diffusion. Hence, the hydrogels were noncytotoxic. Moreover, cell-material interaction studies show that these hydrogels were nonadhesive. Finally, histologic evaluation of tissue response to subcutaneous and intramuscular implants showed acceptable levels of biocompatibility, as characterized by a normal cellular response and the absence of necrosis of the surrounding tissues of the implant. In the first 10 days, the foreign-body reaction in the intramuscular implantation was more severe than in subcutaneous implantation, becoming identical after 30 days. In both cases, dextran hydrogels did not show signs of degradation 6 weeks postimplantation and were surrounded by a thin fibrous capsule and some macrophages and giant cells. This response is typical with a number of nondegradable biocompatible materials. These results indicate that dextran hydrogels are biocompatible, and may have suitable applications as implantable long-term peptide/protein delivery systems or scaffolds for tissue engineering. PMID- 14762940 TI - Management of positional skull deformities: who needs a helmet? PMID- 14762941 TI - Management of positional skull deformities: who needs a helmet? PMID- 14762942 TI - Indiana uses 'assemble' approach to tackle chronic disease. PMID- 14762943 TI - Comprehensive approach to depression management is put to the test. AB - With depression topping the list of behavioral health issues affecting its members, a large Midwestern insurer has turned to a newly developed approach to depression that includes risk-stratification, several levels of intervention, and a philosophy that helps members feel more empowered to help themselves. There are early signs this approach is delivering solid clinical improvements, and developers are hoping these will translate into a strong ROI as well. PMID- 14762944 TI - New, state-of-the-art center has a laser-beam focus on prevention. AB - How often do you see patients who appear headed for a chronic disease, but you feel powerless to intervene? A new center has opened its doors to provide alternatives to providers who want to establish their patients on a path toward prevention rather than disease. The approach combines state-of-the-art technology with the latest science on prevention, and it comes at a price. However, developers believe it is a price individual consumers should be willing to pay. PMID- 14762945 TI - New approach adds risk-reduction, lifestyle management to traditional DM services. PMID- 14762946 TI - Recovery position. PMID- 14762947 TI - Clinical management. Where medicine meets management. Destination unknown. AB - A south-east London GP co-operative has expanded beyond provision of out-of-hours care. Skill mixing is to be introduced to free up GPs' time. Pilots in north-east London are working on continuity of care. PMID- 14762948 TI - Organisational theory. All fall down. PMID- 14762949 TI - What works. The youth vote. PMID- 14762950 TI - Mental health. Growing pains. PMID- 14762951 TI - Law. For medicinal use. PMID- 14762952 TI - HSJ people. Back in the loop. PMID- 14762953 TI - Medical gas pipeline systems--HTM 2022 revision. PMID- 14762954 TI - True partnership approach excels. AB - Partnerships and alliances will be essential for success in delivery of lower value projects as well as high-value ones, Graham Elderfield, chief executive of the Isle of Wight Healthcare NHS Trust, maintains. He described, at the latest Healthcare Estates Conference, the highly successful, rapid provision of a new acute mental health unit on the Island, and the development is reviewed in this article. PMID- 14762955 TI - Championing innovation and excellence. PMID- 14762957 TI - Glass-fronted focal point. PMID- 14762956 TI - Delivering success in Milton Keynes. AB - The national roll out of the new procurement route, for publicly funded health schemes over 1 million Pound, has recently been implemented and NHS Trusts are quite naturally looking to the pilot regions for a demonstration of what can be achieved. One such example is the successful delivery of the new Treatment Centre for Milton Keynes General Hospital NHS Trust by Norwest Hoist Construction and its supply chain. PMID- 14762958 TI - IT provision faces partnership challenges. PMID- 14762959 TI - Supplies must be sustainable and economical. PMID- 14762960 TI - Equipment expertise extended. PMID- 14762961 TI - In-built protection can help cut HAIs. PMID- 14762962 TI - Baptist Memorial addresses national shortage of emergency room space. Campaign by Memphis, Tenn., system points to minor med facilities. AB - At Baptist Memorial Health Care, Memphis, Tenn., patients expressed some discontent with emergency room procedures. Baptist's public relations team addressed issue, along with the national crisis in emergency room overcrowding. They developed a public service campaign to help the public make correct choices. The campaign also included an education program about St. Mary's ER procedures and the development of a specialized ER volunteer corps, all aimed at having positive long-term results. PMID- 14762963 TI - Sutter Health and PacifiCare work together for co-op advertising. First-time collaboration for hospital and insurer. AB - When Sutter Health, based in Sacramento, Calif., teamed up with PacifiCare Health Systems, the result was a strong combination of the strengths of both. PMID- 14762964 TI - Elegant dinner raises $500,000 for St. Mary's endowment fund. Green Bay Packers attend first event at restored Lambeau Field. AB - A fundraiser for St. Mary's Hospital Medical Center, Green Bay, Wis., received a huge public response. Inspired and facilitated by Madeline Harlan, wife of Green Bay Packers president and CEO, Bob Harlan, it was the first event in the Packers' new Lambeau Field. PMID- 14762965 TI - Sky Ridge Medical Center--first in a fast-growing area. The 'dream job' of promoting a new medical center. AB - The first hospital in rapidly-growing Douglas County, Colo., Sky Ridge Medical Center, Lonetree, Colo., offers luxurious amenities ... and a publicist's dream. PMID- 14762966 TI - Henry Ford Health System's web site has comprehensive information. AB - Henry Ford Health System, Detroit, has won awards for its web site. We bring you some of the highlights of this comprehensive, attractive site. PMID- 14762967 TI - 'We kept caring. And so did you' Crouse's media/PR campaign. Announcing exit from Chapter 11 protection. AB - Crouse Hospital, Syracuse, N.Y., survived a bankruptcy filing and resumed operation on a firm financial basis October, 2003. Read how it publicized its recovery and thanked its loyal employees, staff and volunteers in a comprehensive media/public relations campaign. PMID- 14762968 TI - Marietta Memorial features fine services close to home. Annual report also has community, fund-raising functions. PMID- 14762969 TI - No evidence for an impact of selenium supplementation on environment associated health disorders--a systematic review. AB - In addition to vitamin C (and other vitamins/antioxidants), clinical ecologists (functional medicine) recommend selenium supplementation as a fundamental therapeutic remedy for the treatment of environment associated health disorders. This recommendation is based on the postulation that the trace element selenium inhibits oxidative stress generated during endogenous detoxification of xenobiotics (phase 1) by increasing selenium-dependent glutathione peroxidase activity, and that it counteracts heavy metal toxicity by forming inert metal complexes. The objective of this review was to investigate whether there are any valid studies providing reliable evidence of the therapeutic benefits of selenium supplementation in potentially environment associated health disorders. A systematic review was conducted based on the rigorous and well-defined methods developed by the Cochrane Collaboration. To achieve the demanding standards for systematic review set by the Cochrane Collaboration, study selection, quality assessment and data abstraction were performed independently and in duplicate using a standardized protocol. Overall, 1290 studies were identified as being eligible for inclusion. Twelve of these met the inclusion criteria and their quality was evaluated individually. None of the studies included in the analysis provided evidence of the therapeutic benefits of selenium supplementation in environment associated health disorders. PMID- 14762970 TI - Internal exposure of nursery-school children and their parents and teachers to di(2-ethylhexyl)phthalate (DEHP). AB - Di(2-ethylhexyl)phthalate (DEHP) is the main plasticizer for polyvinyl chloride (PVC) products. It has become widely spread in our environment and among people. DEHP is suspected to be responsible for endocrine-disruptor-like effects in mankind. Children are probably most susceptible to these endocrine effects. In this study we determined the internal exposure of nursery school children (aged 2 6 years) to DEHP and compared it to their parents' and teachers' exposure. The DEHP-metabolites mono(2-ethyl-5-hydroxyhexyl)phthalate (5OH-MEHP), mono(2-ethyl-5 oxo-hexyl)phthalate (5oxo-MEHP) and mono(2-ethylhexyl)phthalate (MEHP) were determined in first morning urine. The sum of the three DEHP metabolites in children's and in adults' urine was 90.0 and 59.1 micrograms/l respectively (median values; p = 0.074). Concentrations of the secondary metabolites 5OH-MEHP (median: 49.6 vs. 32.1 micrograms/l; p = 0.038) and 5oxo-MEHP (median: 33.8 vs. 19.6 micrograms/l; p = 0.015) were significantly higher in children than in adults. MEHP concentrations were low both in adults and children (median: 6.6 micrograms/l vs. 9.0 micrograms/l). Creatinine adjusted values should more accurately reflect the dose taken up with respect to body weight when comparing children with adults. Total creatinine adjusted DEHP metabolites in urine were significantly higher in children than in adults (median values: 98.8 vs. 50.9 micrograms/g creatinine; p < 0.0001). This also applied to the concentrations of both secondary metabolites 5OH-MEHP (55.8 vs. 28.1 micrograms/g creatinine; p < 0.0001) and 5oxo-MEHP (38.3 vs. 17.2 micrograms/g creatinine; p < 0.0001). Creatinine corrected concentrations for the monoester MEHP in children and adults were very similar (8.7 vs. 8.6 micrograms/g creatinine; p = 0.908). Based on the sum of the three determined metabolites we estimated the DEHP dose (in microgram/kg body-weight) taken up by children to be about twice as high as the dose taken up by adults. Routes of the ubiquitous exposure to DEHP remain indistinct. In children's urine the mean relative ratios of MEHP to 5OH-MEHP to 5oxo-MEHP were 1 to 7.1 to 4.9, in adults they were 1 to 3.4 to 2.1. This might indicate an enhanced oxidative metabolism in children. To date no information on the biological activity and toxicity of oxidative metabolites of DEHP is available. Since these are the major metabolites of DEHP toxicological data on these metabolites is urgently needed. PMID- 14762971 TI - Asthmatic disease among urban preschoolers: an observational study. AB - Asthma is of increasing concern especially in industrialized countries. This cross-sectional study was to assess the influence of spatial and temporal variations in the urban air pollution profile on asthmatic disease. The prevalences presented are based on physician-diagnosed asthmatic and allergic disease data, collected between 1993 and 1995. Seven hundred and thirty-six preschool children (age 2 to 7, mean 5.7 years) of 37 daycare centres in the City of Leipzig participated in the study. Variations were observed in the lifetime prevalences of asthma and allergy with differences in a residential area's ambient pollution profile. Depending on the level of traffic (high or low), children residing in areas with a dominant coal-heating emission profile had more frequently a diagnosis of asthma, 17.5% and 8.8% (95% confidence intervals [CI]: 10.8...23.5 and 5.8...11.6, respectively), as compared to those, living in centrally heated areas 13.4% and 5.8% (CI: 6.6...19.3 and 1.2...9.6, respectively). Allergic disorders occurred more often in areas with a predominantly traffic-associated pollution profile, 14.3% and 9.6% vs. 5.8% and 3.7% (CI: 7.4...20.3 and 6.4...12.5; 1.2...9.6 and 0.2...6.5, respectively). Interestingly, asthmatic disease was not necessarily associated with a clinical history of allergies. Of the children with physician-diagnosed asthma, 83.7% were not reported to have a concurrent diagnosis of allergies nor to show clinical symptoms. This suggests that environmental exposures (i.e., complex pollution mixtures associated with residential coal-heating and/or traffic) may have differentially influenced the phenotypic expression of asthma. A qualitative discussion is presented on the occurrence of "asthma without reported allergies" in Leipzig. PMID- 14762972 TI - Interdisciplinary diagnostics in environmental medicine--findings and follow-up in patients with chronic medically unexplained health complaints. AB - PROBLEM: In patients attributing their chronic, medically unexplained complaints to environmental factors the greatest challenge is to overcome their disabling belief in toxicogenic explanations. METHOD: Patients presenting with health complaints that they attributed to environmental causes in an environmental outpatient department (EOPD) within a university medical center in Germany were studied. An interdisciplinary review of previously diagnosed medical conditions, current clinical consultations, personal risk communication and therapeutic advice is presented. Additionally, patient contentedness, complaint development, and belief in environmental attribution in a follow-up interview are given. RESULTS: The open, prospective study comprises 51 patients reporting more than one complaint. Symptoms had lasted for more than 3 years in 63% of the cases. Seventy percent attributed their complaints to more than one environmental cause. The clinical diagnostic procedure reduced the number of prediagnosed clinical conditions by 50%. Numerous foregoing environmental laboratory analyses had overestimated toxicologically relevant findings. These were not confirmed in 80% (8/10) of the cases. In 8% (n = 4) of the patients a relevant environmental or occupational medical condition was found. A mental or behavioral condition was not considered to have first priority in explaining all complaints in 43% (22/51) of the patients. Among these, mostly respiratory or skin-related diseases were found. All patients contacted participated in a follow-up study after a minimum of 21 months. Sixty-seven percent reported having felt that they were taken seriously, 38% felt better after the beginning of the study, and 45% were no longer certain about the importance of the environmental attribution. Since 83% of the patients with a preceding residential diagnosis of MCS or SBS still believed in environmental causes of their complaints in the follow-up study, we conclude that these prediagnoses appear to be a risk for persisting attribution of the environmental factor. About one third (37%) of these patients with complaints that had not been medically explained by an organic condition during interdisciplinary diagnostics had meanwhile consulted a psychotherapist. CONCLUSIONS: Interdisciplinary diagnostics and scientifically based risk assessment in a specialized clinical center were effective and mostly well accepted by the patients and resulted in reduced attribution of complaints to environmental conditions. No indication was found that patients with complaints not medically explained by organic conditions were managed less successfully by this approach. Considering the high costs that these patients have previously caused, it appears valuable to apply an interdisciplinary diagnostic strategy. PMID- 14762973 TI - Exposure to road traffic enhances allergic skin wheal responses and increases plasma neuropeptides and neurotrophins in patients with atopic eczema/dermatitis syndrome. AB - The effect of exposure to road traffic was studied by sitting on chairs for 30 min beside a road with heavy wheeled traffic. Exposure to road traffic enhanced allergen-induced, but not histamine-induced, skin wheal responses in 26 patients with atopic eczema/dermatitis syndrome, while it had no effect on skin wheal responses in 26 normal subjects. Exposure to road traffic also increased plasma levels of substance P, vasoactive intestinal peptide, nerve growth factor, and neurotrophin-3 in patients with atopic eczema/dermatitis syndrome, while it had no effect on these plasma parameters in normal subjects. Collectively, exposure to road traffic may aggravate allergic diseases by enhancing allergic responses with concomitant increase in plasma levels of neuropeptides and neurotrophins. PMID- 14762974 TI - Qualitative and (semi)quantitative characterization of nasal and skin methicillin resistant Staphylococcus aureus carriage of hospitalized patients. AB - The objective of this study was to systematically investigate the carriage pattern and load of newly identified methicillin-resistant Staphylococcus aureus (MRSA) colonized or infected patients before any decolonization took place. Cultures of wounds (38%), of sputum (16%) or throat (10%) and of urine (10%) most frequently gave the initial positive MRSA result. Samples from nose, forehead, neck, axilla, and groin were obtained to determine the extent of nasal and extranasal colonization. Fifty-six (69%) of the screened patients proved to be MRSA positive at one or more of these sample sites, and 53 (65%) were extranasal carriers. The proportions positive for cultures of the nares, forehead, groin, neck and axilla were 54%, 51%, 38%, 35%, and 28%, respectively. The most sensitive screening method (96% sensitivity) was to take a combination of cultures from the nares, forehead and groin. Out of the 56 patients (100% sensitivity) this combination revealed 10 more MRSA-carriers than testing the nose alone (79% sensitivity). But the number of study patients was relative small. Therefore we cannot give general recommendations for MRSA screening on the basis of these results. For our hospital we concluded to take a combination of three screening samples to detect MRSA-carriers. Beside the MRSA-carriage pattern we report about the quantitative whole-body colonization. Out of 41 patients colonized on the forehead, a median of 20 MRSA/24 cm2 was obtained on contact agar plates. On the neck (n = 28), an identical value was found. The median MRSA levels for the nose (n = 44), the groin (n = 31), and the axilla (n = 23) were 80, 50 and 50 cfu/swab streak. The MRSA load varied widely from 1 to more than 100 colonies per culture. Further studies must show whether the individual number of MRSA cultured from different body sites is relevant for transmission, for acquiring infections or for decolonization efficacy. PMID- 14762975 TI - Lessons learned from "the skeptical environmentalist": an environmental health perspective. AB - Few books about the environment have generated as much heated debate as Bjorn Lomborg's 'The Skeptical Environmentalist: Measuring the Real State of the World', published by Cambridge University Press in 2001. A flavor of the controversy can be gleaned from a series of reviews and rebuttals published in 'Scientific American' (Rennie 2002). In general, most positive reviews appeared in the popular press (e.g., 'The Economist', 'Washington Post Book Review', 'The Wall Street Journal') and most negative reviews appeared in the scientific press (e.g., 'Science', 'Nature', 'Bioscience'). Although 'The Skeptical Environmentalist' (TSE) addresses a number of environmental health issues, voices from the environmental health community have not been prominent among the participants in this debate. Now that the dust from the initial stampede to praise and condemn the book has settled, we will explore lessons to be learned from TSE and the associated debate from an environmental health perspective. PMID- 14762976 TI - Revised and new reference values for some trace elements in blood and urine for human biomonitoring in environmental medicine. AB - Reference values for environmental pollutants related to the German population are established continuously by the Human Biomonitoring Commission of the German Federal Environmental Agency. The reference values for arsenic, cadmium, lead, mercury and platinum in blood or urine were derived from the German Environmental Survey 1998 (adults aged 18-69 years). The reference value for lead in blood was lowered for females from 90 to 70 micrograms/l and for males from 120 to 90 micrograms/l, while the values for cadmium of 1.0 and for mercury of 2.0 micrograms/l in blood remained unchanged. For cadmium in urine the reference value was lowered from 1.5 to 0.8 micrograms/l and for mercury in urine from 1.4 to 1.0 micrograms/l. New reference values were derived for arsenic (15 micrograms/l) and platinum in urine (0.01 microgram/l). Additionally, for nickel in urine a new reference value of 3.0 micrograms/l based on data from the literature was established. Reference values for estimation of the selenium status were summarized from the literature. For aluminium in blood or urine no reference values were derived and the use of human biomonitoring to estimate aluminium exposure in environmental medicine is not recommended. PMID- 14762977 TI - Commentary regarding the article by Koch et al.: an estimation of the daily intake of di(2-ethylhexyl)phthalate (DEHP) and other phthalates in the general population. Int. J. Hyg. Environ. Health, 206, 77-83 (2003). PMID- 14762978 TI - The pathogenetic significance of intestinal Candida colonization. PMID- 14762980 TI - Psychotropic drugs and fatal pulmonary embolism. AB - PURPOSE: To examine the association between the use of psychotropic drugs and fatal pulmonary embolism. METHODS: We conducted a national case-control study of fatal pulmonary embolism. Cases were 75 New Zealand men and women aged 15-59 years who died between 1 January 1990 and 31 December 1998, where the underlying cause of death was certified as codes 415.1, 451 or 453 of the International Classification of Diseases (9th Revision). Four controls, matched for sex and age, were selected from the general practice to which each case had belonged. Information was abstracted from the records of general practitioners, family planning clinics and psychiatric services. Odds ratios and 95% confidence intervals (95% CI) were estimated using conditional logistic regression. The key analyses were restricted to cases (n = 62) and controls (n = 243) without major risk factors for venous thromboembolism. RESULTS: Compared to non-use, the adjusted odds ratio for current use of antipsychotic drugs was 13.3 (95% CI: 2.3 76.3). Low potency antipsychotics appeared to carry the highest risk (odds ratio: 20.8 [95% CI: 1.7-259.0]). The main drug involved was thioridazine. The odds ratio for current use of antidepressants was also increased, at 4.9 (95% CI: 1.1 22.5). CONCLUSIONS: Our results for conventional antipsychotics are consistent with previous studies of non-fatal venous thromboembolism. The finding for antidepressants needs to be replicated in other studies. PMID- 14762979 TI - Adverse pregnancy outcomes associated with maternal enalapril antihypertensive treatment. AB - BACKGROUND: Adverse pregnancy outcomes following the use of angiotensin converting enzyme (ACE) inhibitors, including enalapril, have been reported in descriptive studies. However, no analytical studies on the relationship between the adverse outcomes and enalapril gestational exposures are available. OBJECTIVES: To explore the association between enalapril exposure and adverse outcomes in pregnancy, taking into account other possible risk factors. METHODS: We analyzed a series of all usable cases reported to the FDA between 1986 and 2000 in which enalapril was a suspect drug for the observed adverse outcomes (N = 110). Parameters of exposure and reported outcomes as well as information on potentially confounding variables were systematically abstracted from this series by a single physician. Because exposure to ACE inhibitors after the first trimester of pregnancy had been associated with adverse outcomes in the existing literature, we divided the cases into those exposed in the first trimester only (considered as the baseline group) and cases exposed beyond or after this time. Frequency of reported adverse outcomes in the second group was compared with those in the baseline group; odds ratios were computed, taking account of potentially confounding variables by logistic regression where appropriate. RESULTS: Exposure to enalapril after the first trimester of pregnancy was strongly associated with oligohydramnios and specific adverse outcomes thought to be secondary to reduced amniotic fluid volume (limb deformities, cranial ossification deficits, lung hypoplasia), as well as with neonatal renal failure. The relationship did not change after taking numerous potential confounders into account, including duration of exposure, concomitant drug use, maternal age, concurrent disease, neonatal gender, and gestational age at birth. Such a pattern of abnormalities is considered to be a consequence of the effect of ACE inhibition on fetal renal function that develops after the first trimester. CONCLUSION: The specificity and temporality of the observed adverse manifestations suggest a causal relationship to enalapril exposure. PMID- 14762981 TI - Risk assessment of drugs, biologics and therapeutic devices: present and future issues. AB - PURPOSE: The current US system for detecting adverse effects of therapeutics (drugs, devices and biological products) is suboptimal. This report presents the results of an expert workshop on assessing therapeutic risks. This is the second of five workshops coordinated by the Centers for Education and Research on Therapeutics (CERTs) to address the management of therapeutic risks relative to potential benefits. METHODS: The workshop included academic, industry, government and constituency-based leaders. The focus was on the post-approval phase and procedures in the US, but relevant international issues and attendees were included. RESULTS: Substantial deficiencies in the current US system for risk assessment were delineated. Improving the system will involve research into methods to improve risk assessment, enhancement and consolidation of data handling systems, education of healthcare workers, allocation of financial resources and building of constituencies. CONCLUSIONS: We need leadership on multiple levels for global coordination of risk assessment. We can then begin to fill gaps and produce benefits for industry, health authorities, government agencies, healthcare providers, and most important, the public. PMID- 14762982 TI - Do doctors rely on pharmaceutical industry funding to attend conferences and do they perceive that this creates a bias in their drug selection? Results from a questionnaire survey. AB - PURPOSE: To determine the sources of funding for doctors attending conferences and meetings and the doctors' perception on whether their involvement with the pharmaceutical industry created a conflict of interest or bias in their drug selection. METHOD: A postal questionnaire was distributed to 622 hospital doctors and 515 general practitioners (GPs) working in the Edinburgh area in Scotland, UK. RESULTS: The pharmaceutical industry funded approximately half of the meetings and conferences attended by doctors. Less than 20% of the doctors funded themselves. One-third of the meetings would not have been attended if funding from the industry had not been available. Hospital doctors and GPs had similar views on conflict of interest and bias. A minority of doctors (40%) thought that industry involvement created a conflict of interest but the majority of doctors (86%) thought that it did not create a bias in their own drug selection. CONCLUSIONS: If continuing medical education (CME) for doctors is going to rely on pharmaceutical industry funding in the future, then we need more explicit codes of conduct. It is of concern that while many doctors recognise the potential for the industry to influence their prescribing habits, few recognise that they themselves are susceptible. PMID- 14762983 TI - A population-based study of drug use in the very old living in a rural district of Sweden, with focus on cardiovascular drug consumption: comparison with an urban cohort. AB - PURPOSE: To describe drug use among the elderly, with focus on cardiovascular drugs and regional differences. METHODS: Cross-sectional data from a Swedish population-based study on ageing and dementia were used. In rural Nordanstig, drug-use data for 918 participants, 75 years and older (N1), were collected during the period 1995-1998. The data for 335 participants, 84 years and older (N84+), were compared with 418 subjects of the same age group in urban Kungsholmen (K5), data collected 1997-1998. RESULTS: Over 90% of the participants were using drugs regularly or 'as needed'. The most common were cardiovascular drugs, nervous system drugs and drugs for the alimentary tract and metabolism. Polypharmacy (five drugs or more) was common, especially among the oldest, 46% (N84+) and 50% (K5). ACE-inhibitors were used by only 24% of the N1 participants with heart failure. Significantly fewer of cognitively impaired participants were treated with ACE-inhibitors (OR: 0.44) and beta-blockers (OR: 0.50). Significant regional differences among the oldest old were found, with more cardiovascular (OR: 2.72) and less antithrombotic drugs (OR: 0.43) in the rural 84+ group. CONCLUSIONS: The extensive drug consumption, high prevalence of polypharmacy and regional differences stress the need for rigorous monitoring of drug use in the elderly. The data also indicate undertreatment of some cardiovascular diseases in the elderly with cognitive impairment. PMID- 14762984 TI - Patient approach and experience regarding complementary medicine: survey among hospitalized patients in a university hospital. AB - PURPOSE: To characterize the local population that turns to complementary medicine. METHODS: Participants were selected randomly: Medicine [A] (201 patients); Surgery [B] (100 patients); Control [C] (128 patients). The answers to the first two questions determined whether the questionnaire was to be completed. RESULTS: More females sought complementary medicine treatment, with no significant demographic differences among the groups. Nearly 20% of the subjects had academic education while 43% had completed high school. The results of the 16 significant questions were statistically different when comparing groups C and A (p = 0.025) and B (p = 0.011) respectively. A total of 16%, 12.4% and 9% of the subjects respectively preferred a physician MD as their complementary medicine practitioner and 49% [C], 34% [A] and 29% [B] respectively, favored teaching complementary medicine in medical and nursing schools. CONCLUSIONS: A total of 37% [C], 21% [B] and 27% [A] of the subjects experienced complementary medicine intervention at least once. The percentage of individuals holding academic degrees was higher in the group utilizing complementary medicine than those who did not. Only 21% of C and A groups and 12% of B group knew about adverse reactions to complementary therapies. More than 30% favored 'legalizing' complementary medicine. PMID- 14762985 TI - Adverse drug reactions in a south Indian hospital--their severity and cost involved. AB - PURPOSE: The study was aimed to assess the pattern of occurrence of adverse drug reactions (ADRs) in the local population, severity of reported ADRs and additional financial resource utilisation associated with ADRs. METHODS: This was a prospective, spontaneous reporting study conducted over a period of 7 months by clinical pharmacists. The WHO definition of an ADR was adopted. Each ADR was assessed for its causality by using the WHO Probability Scale. The severity of each reported ADR was assessed using the criterion developed by Hartwig et al. The average cost incurred in treating the ADRs was calculated. RESULTS: A total of 270 suspected ADRs were reported and evaluated from 164 patients. A total of 3.7% of the hospitalised patients experienced an ADR, 0.7% of the admissions were due to ADRs and 1.8% had a fatal ADR. The gastrointestinal system (36.3%) was most commonly involved with an ADR. The drug class most commonly implicated with ADRs was cardiovascular (18.3%). Majority (47%) of the reactions were 'moderate' in severity. The total cost incurred in managing all the reported ADRs was Rs 76,564 (US$ 1595) with an average cost of Rs 690 (US$ 15) per ADR. CONCLUSION: Detection and prevention of ADRs at the earliest is very important as they can cause not only morbidity and mortality but also involve high health care cost in their management. Well-trained pharmacists in the area of ADR detection, reporting and monitoring could prove as an asset in providing better patient care. PMID- 14762986 TI - Benefit-risk analysis: examples using quantitative methods. PMID- 14762987 TI - Pharmacoepidemiology and drug safety. PMID- 14762988 TI - Fly-by-wire. PMID- 14762989 TI - Pediatric exertional heatstroke. PMID- 14762990 TI - Minimizing the dark. PMID- 14762991 TI - Thinking at the speed of sound during a transport: lessons from the wild blue yonder. PMID- 14762992 TI - Gunshot wounds of the cranium or torso: implications for spinal immobilization and airway management. PMID- 14762993 TI - Blood transportation for medical retrieval services. AB - INTRODUCTION: Medical retrieval services resuscitate and retrieve the critically ill and injured. Lifesaving treatment includes the transfusion of red blood cells (RBCs). To ensure patient safety and maximum therapeutic benefit for the patient, the transportation of RBCs for transfusion must occur under optimal conditions. Blood shippers currently in use in Australia are principally polystyrene containers, often selected for low cost, although performance validation is becoming increasingly important. The quality of the blood to the consumer may be compromised by the use of inappropriate shippers and transport systems. METHODS: A prototype of a user-friendly soft pack blood shipper was developed and tested in various climatic conditions over 24 hour periods with the RBC temperature monitored. RESULTS: The blood shipper was validated to maintain blood temperature within an acceptable range (2 degrees to 10 degrees C (35.6 degrees-50 degrees F) for up to 16 hours during transport of ambient temperatures ranging from 8 degrees to 35 degrees C. (46.4 degrees-95 degrees F). CONCLUSION: The blood shipper provides adequate storage for blood products under a range of climatic conditions and time periods adequate for most retrieval purposes. This allows optimal blood transport system for the medical retrieval service, helping to ensure maximum patient safety and therapeutic benefit from transfusion. The validated blood shipper is also important to blood banks and blood consumers for nonretrieval blood transportation. PMID- 14762995 TI - [Lakartidningen will be needed for at least 100 more years! A reliable source which sifts, condense and sums up data]. PMID- 14762994 TI - Comparison of intubation attempts and completion times before and after the initiation of a rapid sequence intubation protocol in an air medical transport program. AB - The purpose of this study was to test the hypothesis that the initiation of a rapid sequence intubation (RSI) protocol would decrease the number of intubation attempts and completion times. In the summer of 2001, an RSI protocol was implemented using succinylcholine and etomidate. Before the RSI protocol, patients were sedated with versed and fentanyl for intubation and then paralyzed with succinylcholine to complete intubation, if needed. Retrospectively reviewed patient transport charts yielded 70 patients in the pre-RSI group and 70 patients in the RSI group. The number of attempts in the RSI group significantly dropped in the pre-RSI group, and the time from medication administration to intubation statistically decreased. In this study, the implementation of an RSI protocol resulted in fewer intubation attempts and more rapid intubations. PMID- 14762996 TI - [A risky start for Lakartidningen]. PMID- 14762997 TI - [An ill-matched couple started Lakartidningen--a sensitive cultural personality and a polemist born to lead]. PMID- 14762998 TI - [Time before 1965. A free forum--no"government organ"]. PMID- 14762999 TI - [Lakartidningen as organ of the Medical Society. Formation of opinion and gradual expansion of news after 1965]. PMID- 14763000 TI - [The great change-over in 1965. From a members' paper to a journalistic product]. PMID- 14763001 TI - [The advertisement is a mirror of history. Progress and curiosities blended]. PMID- 14763002 TI - [The cover is Lakartidningen's "face"]. PMID- 14763003 TI - [A mirror of the time for medicine]. PMID- 14763004 TI - [The national medical journals have cultural obligations, too]. PMID- 14763005 TI - [Lakartidningen as bearer of culture: ethics, history of medicine, humanities]. PMID- 14763006 TI - [The ideal medical journal in the age of IT]. PMID- 14763007 TI - [The concepts of diseases during the 20th century. From chlorosis and focal infection to electricity hypersensitivity, fibromyalgia and burnout]. PMID- 14763008 TI - [100 articles about stomach ulcer during 100 years]. PMID- 14763009 TI - [Great changes in the panorama of infections during the 20th century. Better living conditions more important than vaccines and antibiotics]. PMID- 14763010 TI - [Family planning in Sweden during 100 years. Abortion and contraceptive agents- from law-breaking to reproductive right]. PMID- 14763011 TI - [Treatment of infertility--an unheard-of success. Fertilization in vitro started an almost unbelievable development]. PMID- 14763012 TI - [Development of neonatology: from amusement park entertainment to fetal medicine]. PMID- 14763013 TI - [Affective disorders. Diagnostics, epidemiology and neurobiology--three fields which played an important role for the development of effective therapeutic strategies]. PMID- 14763014 TI - [Case reports of Borrelia infection in Lakartidningen in 1909]. PMID- 14763015 TI - [Quackery--a heated discussion during the whole century]. PMID- 14763016 TI - [Psychiatric legislation must consider the reality. The large group of persons with schizophrenia is stigmatized by the crimes only a few commit]. PMID- 14763017 TI - [Health industry and car industry--cuckoo in the nest]. PMID- 14763018 TI - [Administration in the curriculum!]. PMID- 14763019 TI - [Maternal health services must give cause for and explain routine questions concerning violence and abuse]. PMID- 14763020 TI - [Longevity statistics and PSA in primary health care]. PMID- 14763022 TI - [Do we really sell our soul so easily?]. PMID- 14763021 TI - [Stroke--risk factors and primary prevention. Further studies on the role of air pollutants are needed]. PMID- 14763023 TI - [Volunteers in health care?]. PMID- 14763024 TI - Viability of, and basic fibroblast growth factor secretion by, human peritoneal mesothelial cells cultured with various components of peritoneal dialysis fluid. AB - In patients on long-term continuous ambulatory peritoneal dialysis (CAPD), peritoneal dysfunction is considered to be due to the loss of peritoneal mesothelial cells and to subsequent peritoneal fibrosis and neovascularization. Our aim in the present study was to clarify the role of various components of peritoneal dialysis fluid in the occurrence of peritoneal dysfunction in CAPD patients. We used a cell counting assay and ELISA to study the viability of human peritoneal mesothelial cells and their secretion of basic fibroblast growth factor (bFGF)--which induces peritoneal fibrosis and neovascularization--by cells cultured with various components of peritoneal dialysis fluid. The viability of cultured cells, ranked from highest to lowest by solution type, was bicarbonate (40 mEq/L) > lactate (15 mEq/L) + bicarbonate (25 mEq/L) > lactate (40 mEq/L). Viability also showed a concentration-dependent decrease in the presence of advanced glycation end-products of bovine serum albumin. The bFGF level in the supernatant showed a concentration-dependent increase in the presence of glucose and glycated albumin; bFGF level decreased as the bicarbonate concentration increased. Low levels of glucose, lactate, and glycated albumin, and a high concentration of bicarbonate may preserve the viability of peritoneal mesothelial cells and prevent bFGF secretion. PMID- 14763025 TI - Diffusion correction of sodium sieving applicable in a peritoneal equilibration test. AB - Sodium sieving is a measure of free water transport. However its assessment is disturbed when a large difference exists between sodium concentrations in plasma and in dialysate--that is, when the diffusion rate is high. Based on previous findings concerning similarity in the mass transfer area coefficients (MTACs) of sodium and urate, we developed a model that corrects for high diffusion. The model enabled us to predict the dialysate sodium concentration resulting from diffusion alone at any time point during a dwell. The correction was based on knowledge of the intraperitoneal volume at any time point during the dwell, which can be calculated by using a volume marker (reference method). However, in a peritoneal equilibration test (PET), only the drained volume after 4 hours is available, and urate concentration is not routinely measured. Therefore, our objective in the present study was to investigate whether a diffusion correction using the MTAC of creatinine and the drained volume at the end of the dwell would be as accurate in estimating maximum sodium sieving as the reference method is. We analyzed standardized 4-hour dwells in 28 patients, 19 with stable PD and 9 with ultrafiltration failure. The dialysate consisted of a 3.86% glucose containing solution to which dextran 70 was added as a volume marker. The correlation coefficient between the PET correction method and the reference method was 0.92 in all patients [0.90 in stable patients and 0.95 in the patients with ultrafiltration failure (p < 0.01 for all)]. We conclude that a diffusion correction for sodium can be made using PET data. A diffusion correction yields a better estimate of sodium sieving than does the sole use of the lowest dialysate to-plasma (D/P) sodium irrespective of diffusion rate. PMID- 14763027 TI - Glucose and mannitol have different effects on peritoneal morphology in chronically dialyzed rats. AB - In previous studies, we showed that glucose reduces the morphologic changes in rat peritoneum caused by chronic intraperitoneal administration of 0.9% NaCl solution. In the present study, we set out to determine if the observed results were attributable to hyperosmolarity or to the metabolic effect of the glucose. Intraperitoneal catheters were implanted in 19 rats. The animals were then intraperitoneally exposed twice daily for 30 days to 20 mL 0.9% saline supplemented with either 250 mmol/L glucose (GLU, n = 9) or 250 mmol/L mannitol (MAN, n = 10). Control rats did not undergo catheter implantation or the dialysis procedure (CON, n = 6). At the end of the study, a 1-hour peritoneal equilibration test (PET) using Dianeal 3.86% (Baxter Healthcare SA, Castlebar, Ireland) was performed in every dialyzed rat to analyze peritoneal transport. Afterward, the rats were humanely killed by bleeding, and a semi-quantitative scale was used to evaluate adhesions in the peritoneal cavity. Imprints of the visceral mesothelium and samples of the visceral peritoneum (liver) were then taken and analyzed by light microscopy. The PET results for glucose, urea, creatinine, and total protein were comparable in both experimental groups. We found that intraperitoneal adhesions were more severe in the MAN group (6 rats with adhesions graded > 3) than in the GLU group (only 1 such rat). No difference in peritoneal thickness was observed between the experimental groups (MAN: 54.9 +/- 17.8 microns; GLU: 51.2 +/- 14.5 microns); however, in both experimental groups, the thickness was greater than in the CON group (3.9 +/- 0.6 microns). The density of the peritoneal blood vessels tended to be greater in the MAN group than in the GLU group (0.158 +/- 0.072 vessels/1000 microns 2 vs. 0.085 +/- 0.067 vessels/1000 microns 2, p = 0.0541). No visible blood vessels were evident in the CON group. The density of mesothelial cells was higher in the MAN group than in the GLU group (2456 +/- 333 cells/mm 2 vs. 2090 +/- 322 cells/mm 2, p < 0.05), and, in both experimental groups, the cell density was higher than in the CON group (817 +/- 100 cells/mm 2, p < 0.01). The nucleus: cytoplasm area ratio in mesothelial cells was comparable in the MAN and GLU groups (0.206 +/- 0.039 and 0.176 +/- 0.045), but that ratio was higher in both experimental groups than in the CON group (0.086 +/- 0.010, p < 0.01). We conclude that glucose-induced changes in the peritoneum of rats exposed to chronic peritoneal dialysis depend on both osmotic and metabolic effects. PMID- 14763026 TI - Effects of antihypertensive drugs on peritoneal vessels in hypertensive dogs with mild renal insufficiency. AB - The transport capacity of any membrane depends on its surface area and permeability. In addition, peritoneal capillaries are probably barriers to solute transport. Although no decisive use of antihypertensive drugs has been reported in continuous ambulatory peritoneal dialysis (CAPD) patients with hypertension, those drugs are known to have various effects on vessels. In the present study, we used a charge-coupled-device (CCD) camera in renovascular hypertensive dogs with mild renal insufficiency to investigate the effects of various antihypertensive drugs on the peritoneal capillaries. Renovascular hypertension was induced in the dogs by placing silver clips on both renal arteries to create 90% occlusion. After confirmation of elevation of blood pressure (usually 20 days after the operation), each dog's abdomen was opened while the animal was under general anesthesia. Using a CCD camera, the diameters of the small arteries of the peritoneum were measured after 3 days' oral administration of a placebo (n = 5); or of 8 mg CS866, a selective angiotensin II type 1 receptor blocker (n = 5); or of 10 mg benazepril, an angiotensin-converting enzyme inhibitor (n = 5); or of 10 mg amlodipine, a calcium antagonist (n = 5). In dogs receiving CS866, blood pressure decreased to 128 +/- 6 mmHg from 160 +/- 6 mmHg (p < 0.01). A similar decrease in blood pressure was observed with the use of the other drugs. The diameter of the small vessels increased by 28% +/- 6% in dogs receiving CS866 and by 24% +/- 5% in dogs receiving benazepril, as compared with 3% +/- 3% in dogs receiving the calcium antagonist. These data clearly demonstrate that blockade of the renin-angiotensin system produces an increase in solute clearance in hypertensive dogs with mild renal insufficiency and that such blockade may be applicable as therapy for hypertensive patients on CAPD. PMID- 14763028 TI - Morphologic changes in the liver of dialyzed rats--preliminary observations. AB - Peritoneal dialysis affects both the quantity and quality of connective tissue in the visceral peritoneum. In the present study, we report the alterations observed in the morphology of the superficial liver lobuli of dialyzed rats. The studies were performed in male Wistar rats weighing 250-350 g. Rats were exposed intraperitoneally to 0.9% NaCl, phosphate-buffered saline (PBS), or commercial dialysis solutions containing 3.86% glucose (Dianeal 3.86%: Baxter Healthcare SA, Castlebar, Ireland; CAPD3 and CAPD3-Balance: Fresenius Medical Care, Bad Homburg, Germany) twice daily for 4-6 weeks. At the end of the study, samples of the liver were taken and stained for light microscopy (hematoxylin and eosin, van Gieson). The results obtained in dialyzed rats were compared with those from the control, non dialyzed animals. In control animals, the surface of the hepatic parenchyma was smooth. In all dialyzed rats, irrespective of the solution used, folding of the surface of the liver parenchyma was found owing to penetration of connective tissue elements between the hepatocytes. In effect, folds of hepatocytes within the liver capsule became detached and isolated from the remaining cells of the lobules. The distinctive feature of that pathologic change was that its severity increased with the thickness of the peritoneum. The change was seen in rats exposed to any of the experimental solutions, and therefore appeared to be attributable to a non-specific reaction caused by exposure of the peritoneum to dialysis fluid and not to specific components of the fluid. The observed alterations in morphology seem to suggest disturbed function within the affected lobules. Further studies are necessary to confirm this hypothesis. PMID- 14763029 TI - The effect of dwell time on dialysate cancer antigen 125 appearance rates in patients on continuous ambulatory peritoneal dialysis. AB - The dialysate concentration of cancer antigen 125 (CA125) can be considered a reflection of mesothelial cell mass or turnover in stable continuous ambulatory peritoneal dialysis (CAPD) patients. The effect of dwell times exceeding 4 hours on CA125 appearance rate (CA125AR) is not known. Therefore, our objective in the present study was to analyze the effect of dwell time on CA125AR in stable CAPD patients. In 43 stable CAPD patients, we analyzed standard peritoneal permeability analyses (SPAs) performed with a 3.86% glucose dialysate, and night dwell effluents from the night dwell prior to the SPA. Dialysate CA125 concentration was measured by radioimmunoassay (RIA II: Fujirebio Diagnostics, Malvern, PA, U.S.A.). Night-dwell CA125 correlated with the duration of the dwell (r = 0.32, p = 0.04) and with the CA125 concentration in the 4-hour dwell (r = 0.83, p < 0.001). The mean CA125AR in the SPA effluent was 97.8 +/- 46.3 U/min; in the overnight effluent, it was 108.8 +/- 73.7 U/min (nonsignificant). A good correlation was present between the CA125AR in the 4-hour dwells and in the overnight dwells (r = 0.82, p < 0.001). We conclude that using night dwells to regularly assess dialysate CA125--for instance, at every out-patient visit--is possible in CAPD patients, provided that appearance rate is calculated. PMID- 14763030 TI - Intraperitoneal and subcutaneous pharmacokinetics of low molecular weight heparin in continuous ambulatory peritoneal dialysis patients. AB - Today, low molecular weight heparins (LMWHs) are more and more commonly used. They are about to replace standard heparin in certain circumstances. The pharmacokinetics of intraperitoneal standard heparin are well known in continuous ambulatory peritoneal dialysis (CAPD), but data concerning LMWHs are lacking. The present study investigated the pharmacokinetics of intraperitoneal LMWHs in a single dose and compared them with the subcutaneous route in CAPD patients. The study enrolled 8 CAPD patients with a mean age of 47 +/- 14.14 years. All patients had 40 mg enoxaparin added to their night exchange on one day. Blood samples were drawn just before instillation and at 2, 4, 8, 12, 18, and 24 hours after instillation for determination of plasma antifactor Xa activity. After two days of washout, the same patients were given enoxaparin 40 mg subcutaneously, and blood samples were drawn at the same time points. Although no plasma factor Xa activity was seen after intraperitoneal administration, subcutaneous administration resulted in increased plasma factor Xa activity. We conclude that a single dose of intraperitoneal enoxaparin did not cause any change in plasma anti-factor Xa activity. That finding may be due either to an insufficient dose or to nonabsorption. PMID- 14763031 TI - Clinical experience with tamoxifen in peritoneal fibrosing syndromes. AB - Peritoneal sclerosis is one of the most important complications of peritoneal dialysis (PD) treatment. Encapsulating peritoneal sclerosis (EPS) represents the most advanced stage of that disease and has a high mortality. No therapy of choice has been established for sclerosing peritonitis, although many have been proposed, with variable results. Tamoxifen has been successfully used in the treatment of patients with fibrosing diseases, mainly retroperitoneal fibrosis. Our purpose in the present study was to investigate whether treatment with tamoxifen in PD patients with peritoneal sclerosis has a beneficial effect. Among more than 450 patients treated in our program since 1980, 23 were diagnosed with peritoneal sclerosis. Of those 23.9 were treated with tamoxifen [20 mg every 12 hours: tamoxifen group (TG)] for a mean period of 14.5 +/- 7 months (range: 6-30 months). The other 14 patients received no treatment and were considered the control group (CG). Both groups were similar in demography and peritoneal antecedents. Follow-up was longer in CG than in TG (mean: 47 months vs. 29 months), but the difference did not reach statistical significance. Mild thrombopenia in 1 patient was the only toxic effect observed with the use of tamoxifen. In CG, 4 patients developed EPS and died--3 of them during the first 6 months after diagnosis. No patient treated with tamoxifen developed EPS. Overall mortality was significantly higher in CG (71% vs. 22%, p = 0.03). Although follow up was longer in CG, half the patients in that group died during the first 2 years after diagnosis. Our experience suggests that treatment with tamoxifen of patients diagnosed with peritoneal sclerosis diminishes the related complications and significantly reduces mortality, at least in the short- to mid-term. However, a prospective therapeutic trial is required to confirm our results. PMID- 14763032 TI - Volume expansion and sodium balance in peritoneal dialysis patients. Part I: Recent concepts in pathogenesis. AB - Cardiovascular disease is a leading cause of death in patients with end-stage renal disease. Uncontrolled hypertension and volume expansion contribute to alternations in left ventricular geometry and are independent predictors of poor survival in dialysis patients. Excessive salt intake is a major handicap with loss of residual renal function. Sodium removal becomes inadequate in the face of declining residual renal function. Continued salt intake and inadequate sodium removal lead to volume expansion, which aggravates arterial hypertension. In part I of this two-part review, we consider information on dietary salt intake and its relationship to blood pressure and volume control in peritoneal dialysis (PD) patients. In addition, we review recently published studies on the use of various PD modalities to remove sodium, emphasizing the significance of volume expansion and uncontrolled hypertension in PD patients. Part II reviews the various measures available to enhance sodium and fluid removal in PD patients. PMID- 14763033 TI - Volume expansion and sodium balance in peritoneal dialysis patients. Part II: Newer insights in management. AB - As noted in part I of this article, emerging evidence indicates that overt and (more commonly) subclinical volume expansion is frequent in patients on peritoneal dialysis (PD). That expansion in turn leads to hypertension. With loss of residual renal function, the hypertension becomes difficult to control even with increasing doses and varieties of antihypertensive drugs. Poor volume and blood pressure control aggravates already existing left ventricular hypertrophy and leads to increased cardiovascular morbidity and mortality. Indeed, cardiovascular events are the leading cause of death in PD patients. Part II of this article reviews various strategies available to manage fluid overload and hypertension. Also, we discuss sodium removal with various PD modalities and clinical studies related to use of low-sodium dialysis solutions. PMID- 14763034 TI - Short peritoneal equilibration test: impact of preceding dwell time. AB - In the original peritoneal equilibration test (PET), which established standard values for membrane categorization in 1987, the dwell time of the preceding exchange was approximately 8 hours. Basing the test on an 8-hour prior exchange was convenient when almost all patients were on continuous ambulatory peritoneal dialysis (CAPD); however, many patients are now on some form of automated peritoneal dialysis (PD), and an 8-hour exchange before the PET requires a change in the dialysis schedule. Our goal in the present study was to determine whether the dialysate-to-plasma ratio (D/P) of solutes and the final dialysate to initial dialysate ratio (D/D0) of glucose in the short PET (2-hour equilibration time) are similar whether a 3-hour exchange or an 8-hour exchange precedes the test. The PETs were performed in 9 stable PD patients using 2 L of 2.5% dextrose dialysis solution (Baxter Healthcare, Deerfield, IL, U.S.A.). Before the equilibration exchange, the patients were on a randomly selected long (approximately 8-hour) or short (3-hour) dwell. During the equilibration exchange, dialysate samples were collected at 0.30, and 60 minutes of dwell time and from the drain bag (120-minute sample). Within a week, the PET was repeated in each patient after the alternate-length exchange. The values for D/P creatinine and urea and for D/D0 glucose were almost identical throughout the 2 hour PET after either the long or the short exchange. The D/P protein values tended to be higher in the PET after the long exchange. The short PET can be used for clinical purposes, and the creatinine and glucose results can be used for membrane categorization. Any dwell time between 3 and 12 hours is acceptable for the preceding exchange, and the equilibration test may be performed with either a 2-hour or a 4-hour dwell. The protein values obtained after a 3-hour prior dwell differ from those obtained after a long prior dwell. Protein values must therefore be standardized in a larger number of patients. PMID- 14763036 TI - Dialysate interleukin-15 concentration and ultrafiltration capacity in patients undergoing peritoneal dialysis. AB - Our aim in the present study was to simultaneously determine, in peritoneal dialysis (PD) patients, interleukin-15 (IL-15) concentrations in serum and dialysate, peritoneal transport parameters, and ultrafiltration (UF). The study was performed in 42 patients who had been treated with PD for 24.5 +/- 20.1 months. Concentration of IL-15 was measured (ELISA: IBL, Hamburg, Germany) during a peritoneal equilibration test. Of 42 patients, 69% had serum IL-15 concentrations above normal values (< 13.0 pg.mL). The mean dialysate-to-plasma ratio (D/P) for IL-15 exceeded 1. We observed a negative relationship between dialysate IL-15 and effluent volume. Patients with UF < or = 200 mL or < 100 mL had higher dialysate IL-15 concentration, higher D/P for IL-15, higher mass transfer area coefficient (KBD) for creatinine, and higher dialysate creatinine concentration than patients with more UF. The KBD for creatinine correlated negatively with dialysate effluent volume. We conclude that, in PD patients, elevated serum and dialysate IL-15 concentrations confirm the existence of systemic and local inflammation; the source of IL-15 in dialysate is not only serum, but also local generation in the peritoneum; and dialysate IL-15 concentration may be a marker of UF capacity. PMID- 14763035 TI - A selective angiotensin receptor antagonist, Valsartan, produced regression of left ventricular hypertrophy associated with a reduction of arterial stiffness. AB - We investigated whether a selective angiotensin II receptor blocker (ARB) would have a regressive effect on left ventricular hypertrophy (LVH) in patients on continuous ambulatory peritoneal dialysis (CAPD). In a double-blind study, 24 CAPD patients with LVH [left ventricular mass index (LVMi) > 110 g/m2 for women and LVMi > 137 g/m2 for men] were randomized to 12 months' administration of either the ARB valsartan (n = 14) or a placebo (n = 10). The target blood pressure (BP) was 140/90 mmHg or lower in both groups. The following parameters were measured before and at the end of the study: aortic and large-artery compliance and arterial wave reflections [pulse wave velocity (PWV) and augmentation index (AI) application tonometry] and cardiac echocardiography. Periodically recorded were body weight, BP (mercury sphygmomanometer), serum creatinine, electrolytes, complete blood cell counts, urine volume, drainage volume, and weekly creatinine clearance. Two-way analysis of variance for repeated measurements was used for statistical analysis. Systolic and diastolic BP were both reduced in patients treated with ARB. The LVMi was significantly reduced in patients treated with ARB (to 121 +/- 4 from 145 +/- 5) but not in those receiving placebo (to 137 +/- 3 from 152 +/- 3, p < 0.05). The decrease in LVMi was associated with a reduction in PWV and AI. In CAPD patients with LVH, ARB reduced LVMi in association with alterations in arterial hemodynamics. PMID- 14763037 TI - A partial explanation for the surprising ADEMEX result. AB - Among the many factors contributing to mortality and morbidity in dialysis patients, nutrition is one of the most important. The ADEMEX (Adequacy of Peritoneal Dialysis in Mexico) study suggests that increasing the amount of daily dialysis to compensate for loss of residual renal function (RRF) does not change mortality or morbidity in peritoneal dialysis (PD) patients. Our purpose in the present study was to determine whether a gradual increase in daily dialysis volume to compensate for loss of RRF interferes with the nutrition of patients. We studied the correlation between normalized protein catabolic rate (nPCR) and daily dialysis volume in 150 PD patients. The Student t-test was used to discover if the correlation was statistically significant. We found that, as the daily dialysis volume increases to replace lost RRF, nPCR declines significantly. This reverse relationship was statistically significant at a p value of 0.007. Replacement of lost RRF by an increase in daily dialysis volume in PD patients contributes significantly to their state of protein malnutrition. The large quantity of carbohydrate acquired through dialysis interferes with the patients' intake of protein. The resulting condition of malnutrition probably plays a significant role in mortality and morbidity in those patients. PMID- 14763038 TI - Long-term automated peritoneal dialysis in patients with refractory congestive heart failure. AB - Patients with severe congestive heart failure (CHF), mainly class IV on the New York Heart Association (NYHA) scale, became refractory to standard medical therapy. The factor that favored that evolution was renal insufficiency with inadequate renal perfusion. Our objectives in the present study were to make a preliminary assessment of the usefulness of automated peritoneal dialysis (APD) for the treatment of volume overload in those patients. Our study was carried out in the peritoneal dialysis unit of the clinical hospital of Pontificia Universidad Catolica de Chile. We studied 3 non uremic patients with refractory CHF secondary to ischemic cardiomyopathy, severe secondary lung hypertension (> 70 mmHg), and associated moderate renal insufficiency. The patients (2 women and 1 man) ranged in age from 55 years to 68 years. A Tenckhoff double-cuff peritoneal catheter was placed in each patient, and peritoneal dialysis was carried out using the nightly intermittent peritoneal dialysis (NIPD) modality. The treatment used was appropriate to obtain a gradual removal of fluids: dialysate dextrose at 1.5% and 2.5%, 4-5 cycles, and total volume of 10-12 L per night. All three patients were able to remain at home, with no signs of hypervolemia. The frequency and length of hospitalizations decreased on average from 59 days pre-APD to 37 days post-APD. No hospitalizations for cardiac problems occurred, and the mean survival was 11 months (range: 6-22 months). This preliminary observation suggests that APD could be offered as an effective treatment for helping to remove fluids in patients with refractory CHF, reducing the number and length of hospitalizations, and improving quality of life. PMID- 14763039 TI - Cost savings from peritoneal dialysis therapy time extension using icodextrin. AB - Previous retrospective studies have reported that icodextrin may prolong peritoneal dialysis (PD) treatment time in patients with refractory fluid overload (RFO). Because the annual cost of PD therapy is lower than that of hemodialysis (HD) therapy in Australia, we prospectively investigated the ability of icodextrin to prolong PD technique survival in patients with RFO. We used a computer model to estimate the savings associated with that therapeutic strategy, based on annual therapy costs determined in a regional PD and HD costing exercise. Patients who met standard criteria for RFO and who were otherwise to be converted immediately to HD, were asked to consent to an open-label assessment of the ability of icodextrin to delay the need to start HD. Time to conversion to HD was measured. The study enrolled 39 patients who were followed for a mean period of 1.1 years. Icodextrin significantly increased peritoneal ultrafiltration by a median value of 368 mL daily. It prolonged technique survival by a mean period of 1.21 years [95% confidence interval (CI): 0.80-1.62 years]. Extension of PD treatment time by icodextrin was particularly marked for patients who had ultrafiltration failure (UFF, n = 20), defined as net daily peritoneal ultrafiltration < 1 L daily (mean extension time: 1.70 years; 95% CI: 1.16-2.25 years). Overall, annualized savings were US$3,683 per patient per year. If just the patients with UFF were considered, the savings increased to US$4,893 per year. Icodextrin prolongs PD technique survival in patients with RFO, permitting them to continue on their preferred therapy. In Australia, that practice is highly cost-effective, particularly in individuals with UFF. PMID- 14763040 TI - Improving adequacy of peritoneal dialysis in ESRD Network 5 (Maryland; Washington, DC; Virginia; West Virginia). AB - The goal of the quality improvement project reported here was to increase the proportion of peritoneal dialysis (PD) patients receiving adequate dialysis as defined by the National Kidney Foundation-Dialysis Outcomes Quality Initiative (NKF-DOQI) guidelines. Our approach was to increase the frequency with which we measured PD adequacy and changed prescriptions in response to low adequacy values. We developed 3 indicators, each one subdivided to reflect differences by type of PD and time on PD. Our improvement goal was to achieve the midpoint between baseline performance on those indicators and 100%, equating to a 50% reduction in failure rate (RFR). At baseline, the project included 122 facilities with 1,517 patients (data from October 1999 to March 2000). At re-measurement, we had 117 facilities with 1,372 patients (data from January 2001 to June 2001). In addition to obtaining facility-specific feedback reports, we conducted educational intervention workshops to which all PD providers were invited. After the workshops, "mandatory intervention" facilities submitted improvement plans that were reviewed by the PD Adequacy Work Group to determine if improvement actions were appropriately focused on identified root causes. Not all intervention facilities attended the workshops, and some facilities attended voluntarily. Overall, the Network 5 results showed a statistically significant improvement in measuring PD adequacy (84% baseline to 92% re-measurement), with a corresponding 51% RFR. Improvement in desired levels of adequacy was also statistically significant (55% baseline to 64% re-measurement), with a 21% RFR. Analysis by the intervention group showed that "mandatory intervention" facilities improved more than did "voluntary" facilities in measuring adequacy and in reaching desired levels, and that the differences were statistically significant. Quality improvement efforts that focused on improving the adequacy of PD in Network 5 were accomplished. PMID- 14763041 TI - Incremental dialysis with automated peritoneal dialysis. AB - Since December 2000, we have proposed incremental dialysis to our patients with a glomerular filtration rate (GFR) of 8-9 mL/min/1.73 m2. The incremental dialysis uses automated peritoneal dialysis (APD) 3-4 days per week ["low frequency APD" (LF-APD)], with or without a full abdomen during the period between the nighttime sessions. From December 1, 2000, to September 30, 2001, LF-APD was selected by 5 patients (2 men, 3 women). Of the 5 patients, 4 worked (age: 53 +/- 12 years; range: 40-69 years), and 1 (age: 78 years) needed assistance from a partner who was available only on certain days of the week. Only 1 patient had diabetic nephropathy. The LF-APD was performed by 4 patients on alternate days ("standard LF-APD"), and by 1 patient 3 times per week (owing to limited availability of the partner). At the beginning of treatment, mean nightly LF-APD was 8.9 +/- 0.9 hours with a volume of 13.4 +/- 3.1 L. In 3 patients, icodextrin was used for the dwell between subsequent APD sessions (1-1.5 L). The other 2 patients kept an empty abdomen (owing to body image). Residual renal function (RRF) was measured every 3 months; adequacy and peritoneal equilibration test (PET), every 6 months. Total nightly APD volume was modified based on the results of the PET. At December 31, 2002, 3 patients with follow-ups of 25, 22, and 15 months were still on LF-APD because their GFR had remained unchanged. In another patient, the frequency of the APD sessions was increased after 5 months to 5 times per week from 3.5 times per week following a clinical assessment of underdialysis; however, that patient had had no significant change in GFR. Following a rapid decline in the diabetic patient's GFR (to 3.6 mL/min/1.73 m2 from 7.9 mL/min/1.73 m2 in 18 months) and refusal by that patient to increase the number of days of APD, the diabetic patient was transferred to hemodialysis. No episodes of peritonitis occurred in a total LF-APD follow-up of 84 patient-months. Compliance, assessed using the HomeChoice Pro system and comparing prescribed and recorded sessions, was total (no missed treatments). In conclusion, LF-APD can be a valid form of incremental dialysis and can help patients to choose PD as their first renal replacement therapy. PMID- 14763042 TI - Longitudinal changes of peritoneal function calculated by personal dialysis capacity in a patient after long-term continuous ambulatory peritoneal dialysis. AB - In the present study, we investigated longitudinal changes in peritoneal function, as calculated by the personal dialysis capacity (PDC) test, after patient withdrawal from 17 years of continuous ambulatory peritoneal dialysis (CAPD). In July 1982, a 42-year-old female was started on CAPD because of chronic renal failure. She performed CAPD without any trouble for 17 years. In July 1999, hemodialysis was introduced because of ultrafiltration failure. The CAPD catheter remained in place, and the patient subsequently performed intra-abdominal lavage, using a standard peritoneal dialysis (PD) solution, for 3 years. A PDC test was performed every 6 months before and after withdrawal from CAPD. In July 1999, the PDC test indicated a membrane area of 59,748 cm/1.73 m2, absorption of 2.72 mL/min/1.73 m2, plasma loss of 0.049 mL/min/1.73 m2, and ultrafiltration volume (UFV) of -666 mL/24 h. The dialysate-to-plasma ratio (D/P) of creatinine after 4 hours was 0.96. An abdominal computed tomography (CT) scan showed calcification of the peritoneum. In March 2000, the PDC showed an area of 44,929 cm/1.73 m2, absorption of 1.43 mL/min/1.73 m2, plasma loss of 0.092 mL/min/1.73 m2, and alpha UFV of 213 mL/24 h. In August 2000 (1 year after CAPD withdrawal), the area was 38,492 cm/1.73 m2; the absorption, 1.74 mL/min/1.73 m2; the plasma loss, 0.053 mL/min/1.73 m2; and the UFV, 348 mL/24 h. Long-term intra-abdominal lavage induced a gradual reduction in membrane area, which returned to the normal range (19,028 cm/1.73 m2) after 3 years. Ultrafiltration volume showed a gradual increase and reached its peak level (403 mL/24 h) in July 2002. After 3 years, the D/P creatinine was 0.82. However, an abdominal CT scan showed no change in the calcification of the peritoneum. From those data, we conclude that, in long term CAPD patients, intra-abdominal lavage can induce improvement in peritoneal function, but not in calcification of the peritoneum. PMID- 14763043 TI - Re-evaluation of adequate dose in Japanese peritoneal dialysis patients. AB - Our objective in the present study was to evaluate the dialysis dose required to maintain Japanese peritoneal dialysis (PD) patients in good condition. From 32 local hospitals, we selected 100 stable patients without diabetes mellitus who were on continuous ambulatory peritoneal dialysis (CAPD) or automated peritoneal dialysis (APD), and we performed the peritoneal function test using the newly developed PD NAVI software (JMS, Hiroshima, Japan). Weekly total Kt/V and creatinine clearance (CCr) were calculated and plotted against body weight (BW) to evaluate the PD dose. In CAPD patients (n = 58), we found an inverse linear correlation between total Kt/V and BW (r = -0.576). In the same patients, total Kt remained essentially constant (60 L/week). Those results imply that most patients were being treated with the same PD regimen (2 L, 4 times daily), and that smaller patients were generally receiving a greater PD dose relative to body size than were the larger patients. In APD patients (n = 21), total Kt/V and total CCr normalized to 1.73 m2 did not change with BW. Total Kt and non normalized CCr gradually increased with BW, although the correlation was not significant. Those findings suggest that most APD patients received prescriptions that were more closely based on body size. In conclusion, smaller patients generally receive a greater PD dose than do larger patients, and targeting a single value of Kt/V or CCr may not always be relevant for adequate dialysis. PMID- 14763044 TI - Successful development and application of a strategic dialysis planning model (RENPLAN). AB - Strategic planning for chronic dialysis services to allow cost-effective organization of resources and workforce planning is a difficult but essential task. The planning process within an individual unit involves consideration of both generic issues and issues specific to the unit. We used the Balance of Care approach to planning, which takes into account the affect that the dependency level of patients has on the therapeutic options and on how those options can be delivered. In a workshop format, we developed a spreadsheet computer model (RENPLAN) and applied it to facilitate discussion between clinical and non clinical health care professionals, dialysis unit managers, and renal patient representatives. Within the unit concerned, we used the model to explore a variety of planning options, including reducing the number of patients receiving automated peritoneal dialysis (proposed as a cost-reduction measure by managers). The model precisely calculates future resource requirements (staff numbers, hemodialysis stations, disposables costs), indicates deficits in clinical audit data, and facilitates education of non clinical managers regarding the major issues in dialysis planning. The RENPLAN model is freely available to all units and is adaptable for analysis of local planning issues. PMID- 14763045 TI - Nightly intermittent peritoneal dialysis to initiate peritoneal dialysis. AB - Nightly intermittent peritoneal dialysis (NIPD) with no day dwell has the potential to enhance peritoneal host defenses. We evaluated outcomes in 24 patients who had been placed on NIPD at the start of peritoneal dialysis as compared with outcomes of 24 control patients on standard continuous cycling peritoneal dialysis (CCPD). As compared with patients on CCPD, patients on NIPD had a lower peritonitis rate (0.34 episodes/year vs. 0.59 episodes/year, p < 0.03) and fewer hospital admissions (1.0 admission/year vs. 1.6 admissions/year, p = 0.003). The weekly initial KtV was similar in NIPD and CCPD, but NIPD had significantly higher creatinine clearances owing to higher glomerular filtration rates (GFRs) among the patients (7.8 mL/min vs. 4.8 mL/min, p < 0.02). Technique and patient survival were similar in both groups after controlling for the difference in initial GFR. We conclude that NIPD is a good modality choice for patients starting PD with residual renal function and that it present a low peritonitis risk. As GFR declines, a last fill can be added to maintain adequate clearances. PMID- 14763046 TI - Health literacy affects peritoneal dialysis performance and outcomes. AB - Health literacy (HL) is the ability to perform the basic reading, writing, and numerical skills required to function in a health care setting. Patients with adequate HL are able to read, interpret, and respond to health care information provided by health care providers and health plans. Several means of assessing HL are available for English- and Spanish-speaking patients. A review of the English language literature on HL indicated that no prior studies included a subset of peritoneal dialysis (PD) patients. I administered the Rapid Estimate of Adult Literacy in Medicine (REALM) assessment tool to PD patients. I also asked patients for information about their highest education level completed. Following completion of the REALM, patients were classified as having adequate, marginal, or inadequate HL. As other studies have shown, patients with lower levels of education have inadequate HL. Patients with some college education or higher have adequate HL. However, at the average education level of patients, most patients have marginal HL. Relative lack of HL affects a patient's ability to make decisions regarding care as part of a home self-management program for end-stage renal disease (ESRD) and other chronic illnesses. Consequently, relative HL level affects the method of instruction and the time required for instruction during training of PD patients. PMID- 14763047 TI - Questionnaire to peritoneal dialysis patients undergoing combined therapy (peritoneal dialysis and hemodialysis). AB - Combined therapy [peritoneal dialysis (PD) and hemodialysis (HD)] has been used for ultrafiltration failure and inadequacy of dialysis dose in PD patients. To support the transition to, and continuation of, combined therapy, we investigated patient feelings about combined therapy. We performed a questionnaire survey of 23 combined-therapy patients (mean age: 48.9 years; mean PD duration: 67.9 months; mean combined therapy duration: 21.2 months). The questionnaire asked about feelings before and after combined therapy, physical improvements after combined therapy, and the patient's hopes about the method of dialysis therapy in the future. One patient did not answer, and one questionnaire was removed from the analysis owing to invalid responses. Before combined therapy, 15 patients (71.4%) had negative feelings toward combined therapy. After combined therapy, more than half of the patients experienced improvement in edema, general condition, weight control, complexion, and fatigue. Seventeen patients (81.0%) were satisfied with combined therapy, but 15 patients (71.4%) were uncomfortable with HD. For the future, 15 patients (71.4%) wanted to continue combined therapy, and 5 patients (23.8%) hoped to return to PD only. Patients accepted combined therapy because of the physical improvements, but were uncomfortable with HD because of hospital visits and pain from puncture. Before introducing PD, we need to provide accurate information about PD, combined therapy, HD, and the process of dialysis therapy. PMID- 14763048 TI - Telemedicine system using a cellular telephone for continuous ambulatory peritoneal dialysis patients. AB - We developed a new telemedicine system to monitor the condition of continuous ambulatory peritoneal dialysis (CAPD) patients by using a cellular telephone and an Internet Web site. All data for the CAPD patients--blood pressure, heart rate, body weight, ultrafiltration volume, and urine volume--are collected and sent directly by cellular telephone to a data server that was constructed at the NTT DoCoMo Company data center. The system is directly connected to Internet by application service provider (ASP) technology. Anywhere, at any time, each patient can confirm changes in their data in graph form by using a cellular telephone or a computer connection to an Internet Web site. The average of each type of data is calculated and shown at the Web site. All data collected by cellular telephone are calculated and, in real time, sent directly to the treating physician's office over the Internet. Abnormal data are sent directly to the treating physician's office and shown in the host computer with an emergency signal (emergency alarm system). In addition, CAPD patients can easily contact the medical staff in the Kidney and Dialysis Center of Saitama Medical School (main hospital) using the same telemedicine system. We are using this telemedicine system for 46 CAPD patients being treated by Saitama Medical School. The cost of using the system is just US$3.00 or less per month for each patient. This newly developed system has great advantages for CAPD patients, especially elderly and handicapped patients. The system can be expanded into a network that serves all CAPD patients and all hospitals in Japan. PMID- 14763049 TI - Complications of peritoneal dialysis related to increased intra-abdominal pressure. AB - Peritoneal dialysis (PD) is associated with a number of complications, some of which can be attributed to raised intra-abdominal pressure. Intra-abdominal pressure is highest during coughing or straining--activities which, fortunately, are transient. High pressure, primarily due to high volumes of PD solution, can predispose patients to hernias, dialysate leaks, and back pain; it can also cause altered mechanics of breathing. This article reviews those various complications and their management. PMID- 14763050 TI - Gastric angiodysplasia in patients undergoing maintenance dialysis. AB - We successfully used argon plasma coagulation (APC) to treat two cases of dialysis patients with hemorrhagic gastric angiodysplasia. Gastric angiodysplasia is recognized as an important cause of gastrointestinal bleeding. Angiodysplastic lesion confined to the gastric antrum was first described in 1953 and named gastric antral vascular ectasia (GAVE). The condition is characterized as submucosal capillary dilatation and fibromuscular hyperplasia. The typical finding of GAVE is the so-called watermelon stomach, attributable to vasodilatation. In case 1, a 69-year-old man was introduced continuous ambulatory peritoneal dialysis (CAPD) in July 1997 because of chronic renal failure due to nephrosclerosis. He was hospitalized for severe anemia in December 1997. Gastrointestinal fiberscopy (GIF) showed oozing in the antrum, and gastritis and esophagitis with sliding hernia. Famotidine was started and recombinant human erythropoietin (rHuEPO) was used for anemia. However, the severe anemia did not improve. The patient was hospitalized again for severe anemia and hematemesis. Another GIF showed typical watermelon stomach, which corresponded with GAVE. An APC was performed without complications. Three months later, the anemia was improved, and the dose of rHuEPO was reduced. In case 2, a 57-year-old woman was introduced to hemodialysis in 1998 for uremia due to nephrosclerosis. In October 2000, she was hospitalized for rHuEPO-resistant anemia. A GIF showed oozing in the antrum with diffuse vasodilation in the antrum; GAVE was diagnosed. An APC was carried out without complications. Three months later, anemia was improved. Recently, gastric angiodysplasia was reported to be an important complication in dialysis patients and was recognized as an important cause of rHuEPO-resistant anemia. Argon plasma coagulation is an effective treatment for gastric angiodysplasia in patients on dialysis. PMID- 14763051 TI - Once-weekly hemodialysis helps continuous ambulatory peritoneal dialysis patients who have insufficient solute removal. AB - The most common cause of drop-out from continuous ambulatory peritoneal dialysis (CAPD) therapy is an insufficient dose of dialysis. Several reports and the Dialysis Outcomes Quality Initiative (DOQI) guidelines recommend maintaining a weekly creatinine clearance (CCr) of at least 60 L/1.73 m2. Previously, at our center, we found that many patients switched from CAPD to hemodialysis (HD) owing to insufficient solute clearance (less than 50 L/1.73 m2). We attempted to determine whether once-weekly HD would improve solute clearance. We treated 7 cases (6 men, 1 woman; average age: 54.3 +/- 4.5 years; mean duration of CAPD: 4.3 +/- 1.1 years) with once-weekly HD therapy (3.5 hours; 200 mL/hour). The average CCr was 45 +/- 2 L/1.73 m2. No ultrafiltration failure was found. Addition of once-weekly HD therapy improved CCr to 66 +/- 7 L/1.73 m2. That improvement was attributable to not only to the addition of HD therapy but also to an increase in peritoneal CCr for 3 consecutive days after completion of once weekly HD therapy. Creatinine clearance and ultrafiltration were both significantly increased. Other clinical parameters such as blood pressure control, weight control, and dose of erythropoietin were significantly improved after introduction of once-weekly HD therapy. Moreover, uremic symptoms such as pruritus and depression were markedly improved. In conclusion, once-weekly HD therapy in conjunction with regular CAPD therapy improves solute clearance and symptoms related to uremia in CAPD patients with an insufficient dialysis dose. PMID- 14763052 TI - Peritoneal dialysis versus hemodialysis: a five-year comparison of survival and effects on the cardiovascular system, erythropoiesis, and calcium metabolism. AB - The influence that the mode of dialysis has on the prognosis of patients with renal disease is controversial. The controversy arises at least in part because of the heterogeneity of patient populations, who may be receiving either continuous ambulatory peritoneal dialysis (CAPD) or hemodialysis (HD). In the absence of randomized trials, epidemiologic investigations present the best method for studying the problem. To determine the influence of the mode of dialysis on prognosis and on the cardiovascular system, erythropoiesis, and calcium metabolism, we selected 36 patients undergoing CAPD and 36 patients undergoing HD for a 3-year follow-up study. Patients were matched for age, sex, and cause of renal disease. Among the HD patients, 8 deaths occurred from congestive heart failure, 1 death from cerebrovascular accident, and 2 deaths from severe infection. In the HD group, the average age was 63 +/- 3 years. Among the CAPD patients, 6 were transferred to HD because of recurrent peritonitis or elevation of serum creatinine. Patients on CAPD had lower blood pressures, and patients on HD had lower total cholesterol levels. Other parameters were not significantly different between the two groups, including dose of erythropoietin and calcium supplements administered. Our study provides evidence that clinical outcome in renal failure may depend to some extent on the mode of dialysis. Our results suggest that blood pressure level and serum cholesterol should be taken into account for patients treated with either CAPD or HD. Blood pressure and cholesterol level are both likely to be important contributors to mortality and morbidity in renal patients. PMID- 14763053 TI - Attitudes of Chinese chief nephrologists toward dialysis modality selection. AB - Peritoneal dialysis (PD) is an effective renal replacement therapy; however, its penetration rate in China is rather low. In the present study, we investigated the opinions and attitudes of Chinese chief nephrologists about dialysis modality selection. We asked 51 chief nephrologists from various university hospitals to answer 8 questions about their attitudes toward PD and about the factors that affect their selection of dialysis modality. Hemodialysis (HD) was available in all of the hospitals where the surveyed chief nephrologists worked. Peritoneal dialysis was available in 68.6% of the hospitals, and renal transplantation was available in 54.9%. Of the surveyed nephrologists, 5.9% believed that PD was more advantageous than HD, and 29.4% of them believed that HD was better. However, most of the nephrologists (64.7%) thought that the advantages of each modality depended on the individual patient and therefore that modality selection should be individualized accordingly. About 70.6% of the nephrologists expressed confidence in PD, and 88% were interested in PD. Overall, 90.2% of the nephrologists said that they would start or expand a PD program in the future. More than half of the nephrologists (52.9%) thought that the proportion of PD in China should be 30%-50%. [The actual percentage is less than 10% in most ares (58.8%) of China.] Most nephrologists indicated that the major reasons for such low use of PD were the low profit from PD treatment and insufficient patient and professional education about PD. A few nephrologists said that peritonitis, the high PD dropout rate, and patient preference also affected the penetration of PD. Our findings suggest that many of the chief nephrologists in China are confident about PD treatment. The major obstacles to PD expansion in China are the lack of incentive (profit) for PD treatment and insufficient education for professionals and patients alike. PMID- 14763054 TI - The BDI and the SF36: which tool to use to screen for depression? AB - Recent studies have suggested a relationship between depression--as assessed by the Beck Depression Inventory (BDI)--and mortality in end-stage renal disease (ESRD) patients. A recent study from the Dialysis Outcomes and Practice Patterns Study (DOPPS) indicated an association between mortality in a large cohort of hemodialysis patients and the patients' responses in the preceding 4 weeks to two questions on the Kidney Disease Quality of Life, Short Form (KDQOL-SF36): "Have you felt downhearted and blue?" and "Have you felt so down in the dumps that nothing could cheer you?" A BDI score > or = 11 and a score < or = 3 for the two questions on the SF36 were considered to suggest the presence of depressive symptoms; both scores have been associated with increased mortality in hemodialysis patients. We aimed to examine the relationship of the two SF36 questions with depressive symptoms as assessed by the BDI. All patients on chronic peritoneal dialysis (CPD) therapy and daily hemodialysis therapy in our units between June 2000 and January 2002 were asked to complete a BDI and an SF36. We recorded 135 tests in 80 CPD patients, and 76 tests in 17 daily hemodialysis patients. Correlation coefficients (r2 values) of the responses to the two questions on the SF36 and the BDI scores demonstrated a significant relationship between the scores. The r2 values for the CPD patients' two SF36 responses and the BDI scores were -0.622 and -0.506; the r2 values for the daily hemodialysis patients were -0.363 and -0.317. The sensitivity and specificity for each SF36 response to be < or = 3 when the BDI was > or = 11 were 82.4% and 68.6% for the "downhearted and blue" question and 65% and 67% for the "down in the dumps" question. Whether the two questions on the SF36 that suggest depression can replace the BDI as a screening tool requires further study. Furthermore, it is unclear if the two questions on the SF36 are predictive of mortality because of their association with clinical depression or because of other issues. PMID- 14763055 TI - Promoting use of home dialysis. AB - Despite overwhelming evidence of enhanced survival and quality of life among end stage renal disease (ESRD) patients treated with home hemodialysis (HHD) and home peritoneal dialysis (PD), use of those two modalities is decreasing. Our analysis of data obtained over the past 6 years reveals that, at the same time as the national ESRD population has grown by more than 33%, the fraction of those patients on home dialysis has decreased by 36%. Similar trends are observable in the State of Texas. At our institution, the percentage of ESRD patients receiving home dialysis dropped by 52% in 6 years. Our capture rate during the same interval ranged from 1% to 9% (mean +/- standard deviation: 3.66% +/- 2.64%). To improve the capture rate and to strengthen our home dialysis program, we adopted these measures: Nephrologists participated in dialysis education and explained renal replacement therapy (RRT) options to each patient and to accompanying family members. The home dialysis coordinator later met with the patients individually. The new patients were then encouraged to meet with patients already enrolled in the home dialysis program. Finally, patients choosing home dialysis were given a questionnaire to rank their reasons for selecting that modality. From April 2001 to July 2002, we initiated 136 patients onto dialysis. Of those patients, 118 selected in-center hemodialysis and 18 chose home dialysis, representing a capture rate of 13% and resultant growth of 117% in our home dialysis program. Our survey revealed that the prime reason for selection of home dialysis was independence (31%), followed by physician guidance, coordinator education, and work schedule (17% each). Familial assistance, familial employment, and privacy were less important (7%, 7%, and 3% respectively). We conclude that, by devoting more time to patient education and discussion of RRTs, nephrologists and dialysis coordinators can significantly increase home dialysis enrollment. PMID- 14763056 TI - Histologic criteria for diagnosing encapsulating peritoneal sclerosis in continuous ambulatory peritoneal dialysis patients. AB - To establish histologic criteria for a diagnosis of encapsulating peritoneal sclerosis (EPS), we investigated 69 peritoneal biopsy specimens histologically and immunohistochemically. The specimens included cases of EPS (n = 12), suspected cases of EPS without later manifestation (n = 5), cases of infectious peritonitis (n = 20), cases of ultrafiltration failure (n = 25), and peritoneum at the start of peritoneal dialysis (n = 7). For each specimen, we evaluated these histologic parameters: fibrin deposition, mesothelial denudation, interstitial fibrosis, peritoneal fibroblast swelling, perivascular bleeding capillary angiogenesis, microvascular sclerosis, and interstitial mononuclear cell infiltration. We also evaluated these immunohistochemical markers: macrophage migration inhibitory factor (MIF), fibroblast growth factor (FGF), FGF receptor 2 (FGFR2), alpha smooth muscle actin (alpha SMA), MIB1, and BCL2. The most characteristic histologic findings for EPS were fibrin deposition and fibroblast swelling. The presence of capillary angiogenesis and mononuclear cell infiltration were also associated with EPS. Expression of FGF, FGFR2, MIF, MIB1, and BCL2 in peritoneal fibroblasts was frequently observed in EPS. Our results suggest that fibrin deposition and peritoneal fibroblast activation or proliferation (or both) are useful findings for the early diagnosis of EPS. Careful histologic observation of the peritoneal biopsy after withdrawal of peritoneal dialysis is required for the early diagnosis and prevention of EPS. PMID- 14763057 TI - Encapsulating peritoneal sclerosis-like findings in a hemodialysis patient without a history of peritoneal dialysis. AB - Encapsulating peritoneal sclerosis (EPS) is recognized as a serious complication of peritoneal dialysis (PD). Involvement of inflammation is indispensable as a cause of EPS. Here, we present a case of EPS that occurred in a 46-year-old man with alcoholic liver cirrhosis and ascites who was treated solely by hemodialysis (HD). We managed this patient surgically, with total intestinal enterolysis. Thick capsules had formed surrounding the ascites. Capsules had also covered the parietal peritoneum and intestinal surface and compressed the intestine. Examination of the ascites suggested the involvement of inflammation and the coagulation-fibrinolysis system. Inflammation of a deteriorated peritoneum causes EPS, resulting in encapsulation subsequent to an accumulation of inflammatory products such as fibrin. Even without deterioration of the peritoneum, chronic inflammation and continual irritation caused EPS-like findings with encapsulation. PMID- 14763058 TI - Role of adhesion molecules in the progression of peritoneal sclerosis. AB - To investigate the role of adhesion molecules [intracellular adhesion molecule 1 (ICAM-1), vascular cell adhesion molecule 1 (VCAM-1), and integrin alpha 5 beta 1] in the progression of encapsulating peritoneal sclerosis (EPS) under peritoneal dialysis, we examined changes in the expression of those adhesion molecules in Wistar-Kyoto (WKY) rats treated with acidic dialysis solution with or without angiotensin II type 1a receptor blocker (ARB). We divided 48 WKY rats into 4 groups and dialyzed them with various solutions as follows: (1) pH 7 1.5% glucose dialysis solution (control group, n = 12); (2) pH 3.5 1.5% glucose dialysis solution (EPS group, n = 12); (3) pH 3.5 1.5% glucose dialysis solution, plus oral administration of CS866 5 mg/kg daily (ARB group, n = 12); and (4) pH 3.5 1.5% glucose dialysis solution, plus oral administration of amlodipine (CA group, n = 12). We injected the dialysis solutions into the abdominal cavity and administered the ARB and CA daily for 42 days. On days 3, 7, 14, and 42, three rats in each group were humanely killed by decapitation, and we studied the expression of adhesion molecules in peritoneum by the immunofluorescence method. In the EPS rats, expression of adhesion molecules was observed in peritoneum on day 3 after start of acidic solution treatment, in conjunction with an increment of interleukin 6 (IL-6) in the dialysate. The peritoneum of EPS rats showed peritoneal fibrosis with interstitial cell infiltration. Treatment with ARB significantly suppressed expression of adhesion molecules in the peritoneum and suppressed peritoneal fibrosis. Treatment with a neutral solution induced no peritoneal fibrosis nor expression of adhesion molecules in the peritoneum. Our results suggest that adhesion molecules play an important role in the progression of peritoneal fibrosis and resultant EPS. Treatment with ARB prevents the progression of peritoneal fibrosis and suppresses expression of adhesion molecules in the peritoneum. PMID- 14763059 TI - Peritoneal effluent markers of inflammation in patients treated with icodextrin based and glucose-based dialysis solutions. AB - Chronic exposure to peritoneal dialysis (PD) solutions is associated with a low grade local inflammatory state of the peritoneum. The occurrence of culture negative peritonitis in some PD patients treated with icodextrin focused our interest on subclinical inflammation in icodextrin-treated patients without peritonitis. The aim of the present study was to compare signs of inflammation in icodextrin-treated patients with the same signs in patients using glucose/lactate based (GL) dialysis solutions only. Overnight PD effluents from 19 patients treated with icodextrin and 19 patients treated with GL were investigated for leukocyte count (LC) and differentiation (LD), and for dialysate concentrations of cancer antigen 125 (CA125, the marker of mesothelial cell mass) and hyaluronan (marker of inflammation and tissue remodeling in the peritoneal cavity). Blood cell counts and serum dextran antibodies (DA) were also determined. Total LC in the GL group was significantly lower than that in the icodextrin group. The LD was not different between the two groups, except for the percentage of eosinophils. The blood cell count did not differ between the groups. The median value of DA was similar in both groups. The hyaluronan concentration was markedly higher in the icodextrin group. No significant difference was found for dialysate CA125. In conclusion, the higher effluent cell count, higher percentage of eosinophils, and higher effluent hyaluronan levels in icodextrin-treated patients are consistent with a greater degree of subclinical inflammation during icodextrin treatment than during GL treatment. PMID- 14763060 TI - Severe peritoneal mononucleosis associated with icodextrin use in continuous ambulatory peritoneal dialysis. AB - Icodextrin is a glucose polymer obtained from starch hydrolysis. It is used as an osmotic agent at 7.5% for peritoneal dialysis (PD). Its use in PD has been associated with several side effects separate from the one reported here, the most frequent being sterile peritonitis. Recently, three mechanisms have been proposed to explain the occurrence of sterile peritonitis: allergy to dextrin, production of anti-dextran antibodies, and impurities introduced during manufacture. Here, we report a peritoneal mononucleosis outbreak that is highly suggestive of being a consequence of the last-mentioned mechanism. During the period December 2001 to May 2002, a group of 8 Spanish hospitals whose individual PD programs regularly share information and activity reported 29 cases of sterile peritonitis associated with icodextrin use in continuous ambulatory peritoneal dialysis (CAPD) patients [mean age: 60.7 +/- 14.47 years; 8 women (27.59%), 21 men (72.41%); mean time on PD: 25.21 +/- 35.31 months; mean time on icodextrin: 15.17 +/- 11.03 months]. Of the 29 patients, 51.8% showed no symptoms. The remainder presented with mild abdominal discomfort and anorexia. Only 2 patients showed general malaise, severe nausea, fever, and abdominal pain. The initial white cell count in peritoneal effluent was 512 +/- 386 cells/mL (45.0% +/- 28% neutrophils, 44.92% +/- 32.6% mono-nuclear cells, 7.75% +/- 12% eosinophils). In 5 of the patients, we performed an immunophenotype (CD14) study, demonstrating the monocyte nature of 60%-80% (mean: 70.6%) of the cells. Microbiology cultures were always negative. A rechallenge with the same batches of PD fluid was tried. In 100% of the patients, the clinical and cellular patterns relapsed. No short term changes in peritoneal function have been observed. The manufacturer informed us that the icodextrin was contaminated with a peptidoglycan. In this sterile peritonitis outbreak with a simultaneous, similar clinical presentation in a group of patients treated with icodextrin solution (presumably contaminated with peptidoglycan), clinical outcome was, for the most part, mild-to-moderate. Symptoms disappeared immediately after icodextrin withdrawal and relapsed after rechallenge with the relevant fluid batches. Monocyte cell counts predominated during the episode. Although we cannot rule out an allergic cause, the massive peritoneal mononuclear cell recruitment suggests a particular mechanism. This is a new mechanism for peritoneal cell recruitment in PD. PMID- 14763061 TI - Strategies for intervention in and prevention of biofilm-related infection in peritoneal dialysis patients. AB - Bacteria from the exit site of the peritoneal dialysis (PD) catheter and from contaminated dialysis fluids can grow into microcolonies in biofilm. Those bacteria are implicated in recurrent peritonitis in patients undergoing treatment with PD. The present article highlights new strategies that are designed to alter the major factors regulating biofilm formation and that thereby reduce biofilm related infections in PD patients. PMID- 14763062 TI - The effects of weekly mupirocin application on infections in continuous ambulatory peritoneal dialysis patients. AB - Application of mupirocin to the nares or catheter exit site and frequency of mupirocin administration in continuous ambulatory peritoneal dialysis (CAPD) patients remain controversial. The objective of our study was to evaluate, using a historical control group, the efficacy on CAPD-related infections of once weekly application of mupirocin at the catheter exit site. We instructed 18 CAPD patients, who did not initially use prophylactic antibiotic treatment, about once weekly application of mupirocin ointment to the exit site as part of their exit site care. We recorded the incidence of catheter-related infections, the causative micro-organisms, and the rate of catheter loss. We observed 17 acute exit-site infections (AESIs: 0.45 episodes/patient-year) before mupirocin treatment and 2 AESIs (0.06 episodes/patient-year) after treatment. The relative rate of AESI reduction was 86%. Before application of mupirocin, 52% of AESIs were attributable to Staphylococcus-aureus; after mupirocin administration, no AESIs were staphylococcal. Peritonitis episodes were also reduced from 21 before mupirocin treatment (0.56 episodes/patient-year), to 9 after mupirocin administration (0.29 episodes/patient-year). The relative rate of peritonitis reduction was 48%. Once-weekly application of mupirocin to the exit site resulted in a reduction in exit-site infections and peritonitis episodes comparable to those obtained with daily application. PMID- 14763063 TI - Fivefold reduction in peritonitis using a multifaceted continuous quality initiative program. AB - Despite recent advances in peritoneal dialysis (PD) systems, peritonitis is a significant clinical problem in patients on PD. Risk factors for peritonitis are identifiable and modifiable and require focused intervention. During a baseline period in 1998, we observed consistent differences in peritonitis rates among patients using various PD connection systems. In January 1999, motivated by a need to reduce peritonitis, we initiated a multifaceted continuous quality initiative (CQI) program that included retraining all current patients and all new patients 6 months after initiation and then annually; changing from plastic to titanium adapters between the catheter and the transfer set; and using equipment from a single PD manufacturer for all new patients and for current patients with high peritonitis rates. Furthermore, all patients using HomeChoice cyclers (Baxter Healthcare Corporation, McGaw Park, IL, U.S.A.) were taught to use the Compact Exchange Device II to avoid contamination when spiking solution bags. Peritonitis rates improved from 1 episode per 7.5 patient-months (over 512 patient-months) in 1998 to 1 episode per 36.5 patient-months (over 292 patient months) as of September 2002. Further analysis also showed a significant difference in peritonitis rates between equipment produced by various manufacturers. There was a statistically significant difference in peritonitis for automated peritoneal dialysis systems. Patients using the Freedom Cycler PD+ (Fresenius Medical Care, Frankfurt, Germany) had an average peritonitis rate of 1 episode per 6.9 patient-months as compared with patients using the HomeChoice cycler (Baxter Healthcare), who experienced 1 episode of peritonitis per 23.9 patient-months on average (p < 0.0001). For continuous ambulatory peritoneal dialysis patients using UltraBag (Baxter Healthcare), the peritonitis rate was 1 episode per 26 patient-months as compared with the Premier Double Bag (Fresenius Medical Care), for which a peritonitis rate of 1 episode per 6.3 patient-months was seen (p < 0.0001). Comparison of the UltraBag (1 episode per 26.0 patient months) with the Disposable Freedom Set, a single-bag "Y" system (Fresenius Medical Care; 1 episode per 7.5 patient-months) yielded similar results (p < 0.0001). We conclude that ongoing CQI efforts can significantly reduce peritonitis rates. Our efforts included aggressive patient retraining, protocol changes, use of a titanium adapter between the catheter and the transfer set, and careful choice of connectology systems (possible supplier-dependent effect). PMID- 14763064 TI - Normalization of protein nitrogen appearance by various size indicators in patients on continuous peritoneal dialysis. AB - Protein nitrogen appearance (PNA) estimated from urea nitrogen appearance (UNA) reflects dietary protein intake and is an important index of nutrition in peritoneal dialysis (PD). Normalization of PNA by several body size indicators has resulted in discrepancies between normalized PNA (nPNA) values and other nutrition indices in PD patients with varying weight status. To test whether a particular size indicator produces nPNA values that eliminate the discrepancies, we normalized PNA values obtained at the first clearance study in 925 PD patients by weight (W), by fat-free mass (FFM) obtained from body water, by lean body mass (LBM) derived from creatinine kinetics, by desirable weight (DW), by standard weight (SW), and by adjusted edema-fee body weight (aBW). We classified patients into three groups according to W/DW: an underweight group [group I: W/DW < 0.9, n = 147 (15.9%)], a normal-weight group [group II: 0.9 < or = W/DW < 1.2, n = 506 (54.7%)], and an obese group [group III: W/DW > or = 1.2, n = 272 (29.4%)]. The UNA and PNA increased progressively from group I to group III. The W, FFM, LBM, SW, and aBW also increased progressively, and the corresponding nPNA values decreased progressively from group I to group III--all at p < 0.001. The DW did not differ significantly between the three groups. The PNA values normalized by DW were 0.91 +/- 0.21 (group I), 0.96 +/- 0.24 (group II), and 1.07 +/- 0.25 (group III), p < 0.001. Of the six size indicators tested, only DW produced values of nPNA consistent with the nutrition status of PD patients with varying weight status. Our findings indicate that DW is the proper size indicator for normalizing PNA in PD patients. PMID- 14763065 TI - Body composition evaluation in peritoneal dialysis patients using anthropometric formulas estimating body water. AB - In adult patients on peritoneal dialysis (PD), estimates of body water (V), fat free mass (FFM), and body fat (BF) can be obtained by using anthropometric formulas for estimating V. Estimates of V, FFM, and BF can also be obtained by using reference (standard) methods in which V is evaluated by a standard dilution method. To test whether the estimates obtained by the various methods agree, we analyzed published studies that measured V by a standard method in adult PD patients. We then calculated V, FFM, and BF by the Watson, Hume, Sahlgrenska, Chumlea, Lee, and Chertow formulas for the "average subject" in the published studies. We compared the standard and the anthropometric estimates using the limits-of-agreement method of Bland and Altman. Our analysis included six studies involving a total of 262 patients (89 women, 173 men). The six studies measured V by a reference method and allowed calculation of anthropometric V, FFM, and BF for the average patient. We calculated these values: VStandard, 36.8 +/- 4.7 L; VSahlgrenska, 36.9 +/- 4.3 L [p = nonsignificant (NS); Bland-Altman limits of agreement: -3.1 L to 3.3 L]; FFMStandard 50.9 +/- 5.2 kg; FFMSahlgrenska, 50.6 +/ 4.3 kg (Bland-Altman limits of agreement: -5.4 kg to 4.8 kg); BFStandard, 20.3 +/- 3.0 kg; BFSahlgrenska, 20.6 +/- 2.3 kg (Bland-Altman limits of agreement: 4.8 kg to 5.4 kg). Results obtained using the Watson, Hume, Chumlea, and Lee formulas also agreed with the standard estimates. The Chertow formula produced results that systematically overestimated the standard V and FFM values and that underestimated the standard BF. The Watson, Hume, Sahlgrenska, Chumlea, and Lee anthropometric formulas used to calculate V provide estimates of body composition that approximate those obtained using standard methods in PD patients with average body composition. Anthropometric estimates may cause large systematic errors in PD patients whose body composition varies from the average, particularly obese or overhydrated patients. PMID- 14763066 TI - Weight status classification of patients on continuous peritoneal dialysis. AB - We tested the agreement between classifications of the weight status of patients on peritoneal dialysis (PD) by body mass index (BMI) and by body fat (BF) content (BF/W, where W = actual weight), when BF was computed as BF = W - V/0.73 from the Sahlgrenska, Watson, or Hume anthropometric formulas estimating body water (V) in 933 patients on PD and 7,737 outpatients without hydration disorders. We used currently accepted cut-off values for classifying subjects as underweight, normal weight, overweight, and obese by BMI and BF/W. We obtained these values: BMI: men on PD (n = 555), 25.5 +/- 4.3; men with normal renal function [NRF (n = 5,906)], 27.7 +/- 5.1; women on PD (n = 378), 25.9 +/- 6.1; women with NRF (n = 1,831), 28.3 +/- 6.5; BFSahlgrenska/W--men on PD, 0.238 +/- 0.063; men with NRF, 0.274 +/ 0.052; women on PD, 0.342 +/- 0.89; women with NRF, 0.366 +/- 0.075. We obtained these regressions: Women on PD BMI = 12.0832 + 38.9550 (BFSahlgrenska/W) -92.9252 (BFSahlgrenska/W)2 +254.0675 (BFSahlgrenska/W)3, r2 = 0.917; Men on PD BMI = 19.4729 - 29.1310 (BFSahlgrenska/W) +213.7045 (BFSahlgrenska/W)2, r2 = 0.888. From those regressions, the BMI value corresponding to the BFSahlgrenska/W cut off for underweight was similar to the National Institutes of Health (NIH) BMI cut-off for underweight. The BMI value corresponding to the BFSahlgrenska/W cut off for obesity was substantially lower than the NIH BMI cut-off for obesity. The kappa ratios of the classifications of weight status by BMI and BFSahlgrenska/W varied between 0.142 and 0.304 (poor agreement), with more than 50% of the subjects classified in a more obese weight category by BFSahlgrenska/W than by BMI. Classification of the subjects by BMI and by BFSahlgrenska/W in quintiles or quartiles led to much higher kappa ratios, particularly in women. The results were similar in subjects with NRF and with the use of the Watson and Hume formulas to estimate BF. The use of arbitrary cut-off values of BMI or anthropometric BF/W to classify PD patients or patients without edematous states as underweight, normal-weight, overweight, or obese leads to substantial disagreement between the two classifications. Classification of weight status by BMI or BF/W in quintiles or quartiles improves substantially the agreement between the two classifications and should be preferred. PMID- 14763067 TI - Differences in clinical and laboratory data of peritoneal dialysis patients selected according to body mass index. AB - The aim of the present study was to compare parameters of dietary intake, nutritional status, dialysis adequacy, and laboratory data between continuous ambulatory peritoneal dialysis (CAPD) patients having a normal body mass index (BMI) and those having an increased BMI. Two groups were selected: BMI = 20-25 (group I, n = 29) and BMI = 25-30 (group II, n = 19). No difference in age or peritoneal dialysis duration were observed between the study groups. Among parameters of dietary intake, only vitamin C ingestion was higher in group II than in group I. As compared with the patients in group I, the patients in group II showed significantly higher total body mass, lean body mass, fat body mass, and total body water. The Kt/V was lower in group II than in group I. In group II, values of serum iron concentration and transferrin saturation were both lower than those seen in group I. No differences were observed between the study groups for the other examined parameters. Those results show that overweight CAPD patients have higher anthropometric parameters and a lower Kt/V owing to a higher urea distribution volume. Disorders in iron parameters are more frequent in overweight patients. The observed differences do not influence duration of peritoneal dialysis treatment. PMID- 14763068 TI - The effect of intraperitoneal 22-oxacalcitriol on secondary hyperparathyroidism in continuous ambulatory peritoneal dialysis patients (IPOX study). AB - Secondary hyperparathyroidism is a major complication in uremic patients undergoing dialysis. Various active metabolites of vitamin D are used as oral treatment; however, in some patients, parathyroid hormone (PTH) is not ideally suppressed. Intravenous injection of an active form of vitamin D inhibits PTH more effectively than does oral administration. However, intravenous administration is often restricted by the complication of hypercalcemia. In the calcitriol analog 22-oxacalcitriol (OCT), the 22nd carbon atom is replaced by oxygen. The OCT analog has been reported to have a lesser hypercalcemic effect than does calcitriol. The present study was planned to determine whether intraperitoneal administration of OCT would be a more effective treatment of secondary hyperparathyroidism than intravenous administration is in CAPD patients who manage themselves at home. The results showed that OCT is stable in dialysis solution and that its intraperitoneal administration was effective for suppressing PTH in patients with secondary hyperparathyroidism. PMID- 14763069 TI - Severe hyperparathyroidism despite paricalcitol therapy: one-year follow-up. AB - Paricalcitol [Zemplar: Abbott Laboratories, Abbott Park, IL, U.S.A.] is efficient for treating secondary hyperparathyroidism in patients on maintenance hemodialysis (HD). Zemplar is thought to be more potent than calcitriol and has been reported to cause less hypercalcemia and hyperphosphatemia. Here, we report a 1-year follow-up on patients from one inner-city dialysis unit. We reviewed the charts of 100 patients and collected data for 1 year. Patients were stratified into four groups depending upon their intact parathormone (iPTH) levels. Hemoglobin (Hb) and erythropoietin (EPO) doses were determined. More than 50% of patients had iPTH levels greater than 300 pg/mL. Mean Ca and PO4 levels were not significantly different, but Zemplar doses were significantly different in all groups. None of the patients had symptomatic bone disease. Seven patients were changed to low-Ca dialysate (1.0 mEq/L) secondary to hypercalcemia (Ca > 11.5 mg/dL) and severe secondary hyperparathyroidism. Interestingly, patients with low iPTH (< 100 pg/microL) showed relative EPO resistance, and patients with high iPTH (> 600 pg/microL) required smaller EPO doses. An inverse relationship was observed between Zemplar and EPO dose. The effect of Zemplar on EPO responsiveness needs to be confirmed in a larger study. Our data suggest that severe secondary hyperparathyroidism is frequent despite aggressive paricalcitol therapy in our inner-city HD population. To control severe secondary hyperparathyroidism in these patients, dietary and medication compliance may need to be supplemented with more effective non-calcium phosphate binders or calcimimetic agents, or both. PMID- 14763070 TI - Midpeliq reduces glucose load. AB - The rate of technique failure is still high in Japan for peritoneal dialysis (PD) patients. Of the dropouts who have been treated with PD for more than 6 years, about half suffer from ultrafiltration failure. That condition is supposedly related to the bioincompatible aspects of conventional acidic PD solutions. In 2001, a neutral-pH, lactate-buffered PD solution with low glucose degradation products (GDPs), Midpeliq (Terumo Corporation, Tokyo, Japan), was developed and began to be used in Japan. After switching 3 patients from conventional acidic PD solution to Midpeliq, we observed that 2 patients could then use lower-glucose PD solutions. Case 1 was a 42-year-old woman with a 10-year history of PD. In February 2001, she was switched from Peritoliq (Terumo) to Midpeliq. One month later, she complained of dizziness, and her blood pressure was found to be down to 96/60 mmHg. Post-change fluid removal increased to 1,481 mL from 1,238 mL (p < 0.02). Before the solution switch, this patient exchanged 4 times daily, using 2 L of 2.5% Peritoliq each time. From 3 months after the solution switch, she exchanged 3 times daily using 2 L of 2.5% Midpeliq and 1 time daily using 2 L of 1.35% Midpeliq. Fluid volume removal stayed almost the same. Case 2 was a 52-year old man with a 9-year history of PD. In June 2002, he was switched from Dianeal 4 (Baxter Healthcare, Tokyo, Japan) to Midpeliq. After the change, his daily drainage volume increased from approximately 1,500 mL to 2,000 mL. He began to use 2 L of 1.35% Midpeliq 4 times daily instead of 2 L of 1.5% Dianeal 3 times daily and 2 L of 2.5% Dianeal 1 time daily. At 1 month after the solution switch, his drainage volume was still approximately 1,500 mL daily. Our observations suggest that new, neutral-pH PD solutions such as Midpeliq might reduce the glucose load in addition to having low GDPs and fewer toxic effects on the peritoneum. PMID- 14763071 TI - Malnutrition-inflammation syndrome is associated with endothelial dysfunction in peritoneal dialysis patients. AB - Endothelial dysfunction with atherosclerosis is a recognized complication of uremic patients. The hypoalbuminemia of peritoneal dialysis (PD) patients can induce a hypercoagulable and atherogenic state. In this study, we investigated the role played by malnutrition-inflammation syndrome on endothelial function markers in PD patients. We measured markers of nutrition [normalized protein catabolic rate (nPCR), albumin, prealbumin, insulin-like growth factor 1 (IGF-1), transferrin, and cholesterol], markers of endothelial damage and function [tissue type plasminogen activator (tPA), thrombomodulin (TM), von Willebrand factor (vWF), and NO3 (representing NO)], markers of a coagulable state [fibrinogen and plasminogen activator inhibitor 1 (PAI-1)], markers of inflammation [tumor necrosis factor alpha (TNF alpha) and C-reactive protein (CRP)], and other endothelial injury factors [lipoprotein(a) [Lp(a)] and homocysteine]. We also performed an endothelial stimulation test consisting of right-arm venous occlusion (VO) for 10 minutes. The patients were divided into four groups according to their clinical atherosclerotic score (CAS). We studied 45 clinically stable PD patients. At baseline, statistically significant negative linear correlations were found between albumin and age (r = -0.54, p < 0.05), albumin and vWF post-VO (r = -0.54, p < 0.05), and albumin and TM (r = -0.36, p < 0.05), which are endothelial damage markers and prothrombotic factors. A positive linear correlation was seen between albumin and NO3 post-VO (r = 0.48, p < 0.05), indicating a high vasodilatation capacity. C-Reactive protein and TNF alpha showed a positive linear correlation (r = 0.5, p < 0.01). Similarly, TNF alpha showed a positive linear correlation with cardiovascular risk markers such as fibrinogen (r = 0.79, p < 0.01), PAI-1 (r = 0.44, p < 0.05), and homocysteine (r = 0.37, p < 0.05). Creatinine clearance showed a negative linear correlation with TM (r = -0.36, p < 0.05). Patients with albumin < 4 g/dL showed a lower tPA ratio, lower NO3, and a higher CRP, TNF alpha, and Lp(a) than did patients with albumin > 4 g/dL [tPA ratio: 2.1 +/- 1.56 (n = 29) vs. 2.6 +/- 2.3 (n = 16), p < 0.05; NO3: 47 +/- 27 micrograms/mL vs. 69 +/- 33 micrograms/mL, p < 0.05; CRP: 1.8 +/- 3 mg/dL vs. 1.1 +/- 1.6 mg/dL, p < 0.05; TNF alpha: 44.4 +/- 16 pg/mL vs. 36.6 +/- 21.4 pg/mL, p < 0.05; Lp(a): 55 +/- 39 mg/dL vs. 33 +/- 21 mg/dL, p < 0.05]. Patients with a worse CAS showed higher homocysteine levels and lower albumin values. Those relationships were maintained in both periods of the study. We found no relationships between dialysis dose and endothelial function markers. In conclusion, malnutrition-inflammation syndrome may contribute to endothelial dysfunction and, consequently, to prothrombotic and proatherogenic processes in PD patients. PMID- 14763072 TI - Serum concentration of haptoglobin, adequacy of peritoneal dialysis, and nutrition status of patients with and without diabetes on peritoneal dialysis. AB - The aim of the present study was to establish a relationship between serum haptoglobin (HTG) concentration, peritoneal dialysis (PD) adequacy, and nutrition status in PD patients with and without diabetes. We measured serum concentrations of HTG, albumin, iron, and cholesterol; platelet count; transferrin saturation (TSAT); weekly Kt/V; and total weekly creatinine clearance (CCr) in 60 patients with and without diabetes who were being treated with continuous ambulatory PD or automated PD. The mean serum HTG concentration in PD patients without diabetes (2.5 +/- 1.2 g/L) was elevated and differed significantly from that in PD patients with diabetes (2.0 +/- 1.1 g/L). In patients without diabetes the correlation of serum HTG concentration with serum albumin level was r = -0.330 (p < 0.030), with platelet count was r = 0.320 (p < 0.040), with serum iron concentration was r = -0.450 (p < 0.002), with TSAT was r = -0.4200 (p < 0.005), and with age was r = 0.337 (p = 0.003). No such relationships were seen in patients with diabetes. In both subgroups, no dependence was seen between serum HTG concentration and weekly Kt/V, total weekly CCr, or serum cholesterol concentration. Serum HTG concentration in PD patients without diabetes may be a valid inflammatory marker. The HTG serum level displays a significant statistical dependence on age, platelet count, and markers of nutrition such as serum albumin level, iron, and TSAT. It does not depend on markers of dialysis adequacy (weekly Kt/V, total weekly CCr) or on serum cholesterol concentration. The serum HTG concentration in PD patients with diabetes is lower than that in patients without diabetes, and it is not related to examined factors of inflammation, nutrition, or adequacy of dialysis. PMID- 14763073 TI - Analysis of factors predicting survival of a second peritoneal dialysis catheter. AB - Many studies have examined the survival of the first peritoneal dialysis (PD) catheter. However, data are scarce about the factors that influence the function and survival time of a second PD catheter. The purpose of the present study was to calculate the survival time of the second PD catheter and to examine factors that predict removal of that catheter. We conducted a retrospective study of second PD catheters inserted at our institution over a 10-year period from May 1992 to April 2002. The endpoint consisted of removal of the second catheter. Voluntary change to hemodialysis, kidney transplantation, transfer to another center, and death with a functioning catheter were censored observations. Catheter survival was analyzed by the Kaplan-Meier method. During the study period, 106 patients (59 men, 47 women) received a second catheter. The mean age of the patients was 55 +/- 14.9 years. One third of the patients had diabetes. The reasons for removal of the first catheter and insertion of the second one were peritonitis (n = 50), catheter malfunction (n = 20), catheter leak (n = 11), exit site or tunnel infection (n = 21), and failed kidney transplantation with resumption of PD (n = 4). The median survival of the second catheter was 48 months (95% confidence interval: 38 months to 59 months). On univariate analysis, increasing patient age and peritonitis as cause for removal of the first catheter were associated with an increased risk of removal of the second catheter. However, on multivariate analysis, only increasing patient age was associated with a greater risk to survival of the second catheter. PMID- 14763074 TI - Is it safe to simultaneously remove and replace infected peritoneal dialysis catheters? Review of the literature and suggested guidelines. AB - Since the introduction of Y-connector technology and the subsequent reduction in the frequency of peritonitis, catheter-related infections have become the primary infectious complication in patients on peritoneal dialysis (PD). Such infections may lead to prolonged morbidity, recurrent peritonitis, and catheter failure. Despite appropriate treatment of catheter-related infections, removal of the catheter is sometimes necessary. The timing of catheter removal and replacement has been the focus of significant discussion. The International Society for Peritoneal Dialysis recommends a 3-week interval, but also allows for individualized timing. Long staging periods present problems that simultaneous removal and replacement (SRR) of the catheter may obviate. Here, we review a body of literature on SRR and present guidelines as to when SRR of an infected PD catheter may be considered a safe alternative to a staged procedure. PMID- 14763075 TI - Does endoluminal brushing eliminate the need for catheter removal in peritoneal dialysis patients with persistent peritonitis? AB - Recurrent and persistent peritonitis episodes are exhausting problems in children on chronic peritoneal dialysis (PD) and can lead to discontinuation of treatment. In the present report, we describe our clinical experience with endoluminal brushing (EB) in 3 pediatric patients with refractory peritonitis episodes maintained on chronic PD. The EB was performed on 4 occasions in 3 patients. Peritonitis resolved in 2 of the patients. The remaining patient required removal of the PD catheter. No adverse events have since been observed. Endoluminal brushing should be considered an option for the management of persistent peritonitis before catheter removal. PMID- 14763076 TI - The peritoneal membrane: a dynamic dialysis membrane in children. AB - Peritoneal dialysis prescription in children should be individualized--based not only on numerical targets (Kt/Vurea, Kcreat), but also on consideration of the peritoneal membrane, a dynamic dialysis membrane. In fact, the effective peritoneal surface area is at least a triple entity: an anatomic area, a contact area, and an exchange area. The anatomic area appears to be twice as large in infants as in adults if expressed per kilogram of body weight (BW), although the area is independent of age if expressed per square meter of body surface area (BSA). Therefore, scaling of the intraperitoneal fill volume (IPV) by BSA in square meters is necessary to avoid a low IPV/area ratio, which results in a functionally "hyperpermeable" peritoneal exchange. The contact area (the wetted membrane) is only a fraction of the anatomic area--that is, 30%-60% in humans (by computed tomography). Contact area depends on a variety of factors, such as posture and fill volume, that affect the degree of recruitment of membrane contact area. The exchange area is influenced by both the anatomic are and the contact area. However, it is mainly governed by the specific vascular area as determined by the peritoneal vascular perfusion and the capillaries available for exchange. Vascular area is dynamically affected by a variety of factors, such as the composition of the peritoneal dialysis fluid, the fill volume, and possible inflammatory agents. PMID- 14763077 TI - Outcome in children on chronic peritoneal dialysis. AB - In this retrospective study, we evaluated the causative factors, outcomes, and complications of therapy in 35 patients (16 girls, 19 boys) started on chronic peritoneal dialysis (CPD) between 1997 and 2002. Average age at initiation of CPD was 9.3 +/- 4.4 years. All patients started on continuous ambulatory peritoneal dialysis (CAPD). Nine patients switched to ambulatory peritoneal dialysis (APD) during the follow-up period. The most common cause leading to end-stage renal disease (ESRD) in the patients was reflux nephropathy (22.9%). The major complication during therapy was peritonitis, with 41 episodes seen in 17 patients (1 episode per 18 patient-months). Of the children on APD, 7 developed 17 episodes of peritonitis (1 episode per 8.3 patient-months); of the children on CAPD, 10 developed 24 peritonitis attacks (1 episode per 24.9 patient-months). The other complications were inguinal hernia in 3 patients, subcutaneous leak in 4 patients, dialysate leak in 2 patients, pericardial effusion in 1 patient, umbilical hernia in 1 patient, hydrothorax in 1 patient, and cuff protrusion in 3 patients. During the follow-up period, 4 patients died owing to sepsis or cardiopulmonary complications. Only 1 patient was transferred to hemodialysis (owing to persistent Candida peritonitis). We think that CPD therapy is a good choice of treatment modality in the management of children with ESRD. PMID- 14763078 TI - Adequacy and nutrition in pediatric peritoneal dialysis. AB - Outcomes for pediatric peritoneal dialysis (PD) patients are closely related to dialysis adequacy and nutrition, which need to be measured frequently using a number of laboratory parameters. Although the critical meaning of adequacy and nutrition in the long-term prognosis of dialyzed children is well-documented, PD prescriptions are still largely empirical. Our objective was to evaluate nutritional and dialytic parameters in PD children (urea, creatinine, and albumin excretion in dialysate and urine, and daily protein intake); to measure peritoneal equilibration test (PET) results, Kt/V, normalized equivalent of protein nitrogen appearance (nPNA) and nitrogen balance; and to study the correlations between those variables. We performed 59 prospective laboratory measurements in 15 stable PD patients (7 boys; mean age: 6.7 years; age range: 1.1-14.8 years) during 6 months of follow-up. Creatinine, urea, total protein, and albumin were measured in plasma, urine, and dialysate. We calculated PET, Kt/V, daily dietary protein intake (DPI), protein catabolic rate (PCR), and nPNA. All statistical comparisons used the paired t-test, and correlations were calculated by two-way analysis of variance for repeated measures. A value of p < 0.05 was considered significant. The mean 4-hour dialysate-to-plasma ratio (D/P) of creatinine was 0.78 +/- 0.02 at month 0 and 0.74 +/- 0.13 at month 6 [p = nonsignificant (NS)]. The mean final-dialysate-to-initial-dialysate ratio (D/D0) of glucose was 0.35 +/- 0.11 and 0.34 +/- 0.08 at the same intervals (p = NS). The D/P creatinine showed an inverse correlation with patient age and body surface area, and the D/D0 glucose ratio showed a positive correlation with both of those parameters (p < 0.05). Weekly total and residual Kt/V urea were 3.41 +/- 0.86 and 1.49 +/- 1 respectively. The daily DPI was 3.32 +/- 1.05 g/kg, and the daily PCR was 1.32 +/- 0.47 g/kg, showing a positive net protein balance (DPI-PCR = +2 g/kg daily), which was negatively correlated with age and body surface area (p < 0.001). The mean daily nPNA was 0.94 +/- 0.33 g/kg, which was negatively correlated with age and body surface area (p < 0.05, r = -0.51), and positively correlated with daily DPI and total and residual Kt/V (p < 0.0001). Our patients could be classified as high-average transporters, with low-average ultrafiltration. The high transport state was associated with greater peritoneal albumin losses, a point of concern at younger ages. Total Kt/V and nPNA were higher for the youngest patients, suggesting a favorable nutrition status, but more studies are needed to determine the best value for both parameters in clinical practice. PMID- 14763079 TI - [Medical doctors in private practice in the Greater Paris area. General presentation and overview of results]. AB - The French physician workforce, after decades of growth, will be soon entering a decreasing era. The demographic revolution will affect all parts of the country, including the region Ile de France which has the highest physician/population ratio. An opinion survey conducted among a sample of 1700 physicians gathered 770 replies in March 2003. The difficulties most frequently mentioned by the surveyed doctors are heavy taxes and social contributions (mentioned by 77%), problems for finding locum tenens (52%), traffic jams (48%), irrelevant requests from patients (45%), problems for finding nurses to care patients (28%), conflicts with the sickness fund (24%), lack of security (23%). It is noteworthy that the problems related to locum tenens and nurses are directly associated to the demographic situation of French health workforce. When asked about the risks associated to medical practice, 62% of the surveyed physicians quote the difficulty to combine harmoniously professional practice and family life whereas 60% mention low occupational income. But the highest risk after the surveyed doctors is their legal responsibility (68%). Certain modifications of French regulation concerning medical practice to cope with the forthcoming decrease in the numbers of doctors are approved. In particular, the possibility for a doctor in private practice to hire the services of an other doctor (approved by 41%) and that of having several medical offices (31%). PMID- 14763080 TI - [Characteristics of private practice physicians in the Greater Paris area (Ile de France)]. PMID- 14763081 TI - [Difficulties of professional practice]. PMID- 14763082 TI - [Facing the future, overcoming challenges]. PMID- 14763083 TI - [The physicians from the Greater Paris area (Ile de France) have the floor]. PMID- 14763084 TI - [New European rules for clinical trials benefit Sweden when it comes to competition...a dream country for drug trials]. PMID- 14763085 TI - [People of the whole world have a right to health. Functioning infrastructure and adequate health care are the most important requirements]. PMID- 14763086 TI - [Strongly increasing incidence of adenocarcinoma of the esophagus and gastric cardia]. AB - The objective of this retrospective study was to see whether there was an increasing incidence of adenocarcinoma of the oesophagus and gastric cardia in the Swedish population 1970-1997. If there was, could it be explained as a period or cohort phenomenon? The data were compared with the incidence of squamous cell carcinoma and gastric adenocarcinoma with the gastric cardia excluded. Age standardised incidence for each sex was calculated using the age distribution of the world population as a reference. For the combined group of adenocarcinoma in the oesophagus and gastric cardia incidence gradually increased during the study period. The median increase between adjacent five-year intervals was 14% in men and 20% in women. Previously described risk factors are gastro-oesophageal reflux, obesity and smoking. This study suggests that the increasing incidence also can be explained as a shift in classification from squamous cell carcinoma to adenocarcinoma after 1985. PMID- 14763087 TI - [Difficult to measure results and quality of surgical interventions but length of stay is an important variable]. AB - No generally accepted measures exist to describe the quality of surgical care. Measures derived from surgical care processes are relatively easy to improve but are often too local to allow comparison with other surgical departments. The final outcome of an operation is often difficult to record objectively or can only be determined after observation. Improving items of the process is often insufficient to markedly improve outcome because process and outcome are logarithmically related. Therefore, a department's surgical balance sheet remains stable over several years. The introduction of new techniques or new concepts of treatment with sufficient potential for change can improve that balance sheet if effectively implemented. Quality of care implies a rating of the outcome relative to a standard or in comparison with the achievement of other surgical departments. It is argued that postoperative hospital stay is an objective, verifiable, and uniformly recorded outcome measure that reflects the combined achievement of the surgical, organisational, and social processes for each patient. Laparoscopic cholecystectomy, a new technique, tension-free inguinal hernia repair, and fast-track surgery, a new concept, all had the potential to shorten the hospital stay. The mode of change seems to be diminished surgical and anaesthetic trauma and therefore fewer complications. The ultimate quality achievement is to be able to operate without causing physiological derangement of the patient, which should eventually allow day-surgery for most elective procedures. PMID- 14763089 TI - [Heat shock protein--not only in heat stroke...]. PMID- 14763088 TI - [A rapid development of Achilles tendon rupture following quinolone treatment]. PMID- 14763090 TI - [Complementary food to breastfed infants. Introduction can wait until the age of six months but not longer]. PMID- 14763091 TI - [New map of world health is needed. North and South is changed to healthy and ill and West to rich and poor]. PMID- 14763092 TI - [Health--50 years as a human right. Health care based on rights and non discrimination promotes public health both nationally and globally]. PMID- 14763093 TI - [Primary health care reforms on the international agenda for 50 years. Health for all--now with the focus on results and cost effectiveness]. PMID- 14763094 TI - [Ester Henning--a magic expressionist who created her own world in a psychiatric hospital]. PMID- 14763095 TI - [Collective affiliation to the Medical Society is old-fashioned--time for renewal!]. PMID- 14763096 TI - [ALLHAT--remaining questions await answers]. PMID- 14763097 TI - [Choice of antihypertensive agent--insignificant role of doxazosin (Alfadil). How are evidence-based recommendations implemented?]. PMID- 14763098 TI - [Organ donation and utilitarianism: medical ethics more important than medical efficiency]. PMID- 14763099 TI - Linking team competences to organisational capacities in health care. AB - Palliative care is a complex environment in which teams of healthcare professionals are constantly challenged to match the configuration of care delivery to suit the dynamics of the patient's bio-medical, social and spiritual situations as they change during the end-of-life process. In such an environment these teams need to engage in ongoing interaction between different professional disciplines, incremental improvement in care delivery, learning and radical innovation. This is aimed at combining operational effectiveness, strategic flexibility, exploitation and exploration, in a way that ensures the best possible care for the patient. This paper examines previous research on the management competences and the organisational capabilities necessary for continuous innovation, and analyses evidence emerging from a study of palliative care. Work on the relationships between innovation capacities, organisational capabilities and team-based competence is drawn together. Evidence is presented from research into the management of innovation in palliative care. PMID- 14763100 TI - Care pathways: the road to better health services? AB - This paper describes the development and implementation of care pathways in two orthopaedic units in Scotland. Although originally developed as a tool of project management, care pathways have been promoted internationally as a response to concerns for patient safety, variability in care and increasing costs. Generally, care pathways can be seen as an example of clinician led rather than management led reform. However, it does reflect a wider shift towards process and away from hierarchical approaches to management. Within the UK care pathways have been promoted as a response to the modernization initiative of the Labour Government. While the initiative was a success in both units it was more difficult to implement care pathways in a trauma rather than an elective unit. In conclusion, it is questionable whether care pathways are a universal response to the requirement for modernization and service redesign in the NHS. PMID- 14763101 TI - Achievements of the first wave personal medical services (PMS) pilots in England. A health authority perspective. AB - Personal medical services pilots were introduced in England in 1998 to provide increased flexibility to general practitioners practising in deprived areas, to improve service provision and reduce inequalities. The aim of this study was to identify health authority perspectives of the achievements of their pilots. Less than half of the health authorities agreed that their pilots' original objectives had been completely achieved. Support, commitment and enthusiasm from within and outside the pilots, and the ability to be flexible, were helpful in promoting change management. Obstacles were financial difficulties and a lack of understanding of personal medical services. The opinion was that personal medical services had made a highly regarded contribution to the local health economy, especially in the provision of new services and the promotion of new staff roles. The results provide lessons for primary care organisations in England and elsewhere in terms of the factors required to successfully implement change. PMID- 14763102 TI - Developing the effectiveness of primary care organisations in the UK National Health Service. A case study. AB - This paper aims to explore the early experiences of a new primary care organisation in the NHS. It reports the findings of a longitudinal qualitative case study of one primary care group in its first year of operation. It concludes and makes recommendations in four key areas relevant to the development of the primary care group: the experiences of individuals and their readiness for change; clarity and consensus about roles and responsibilities in the new organisation; the process of change, and the impact of culture/power structures; and developing relationships with internal and external stakeholders. PMID- 14763103 TI - Do star ratings really reflect hospital performance? AB - The publication of performance ratings for NHS Trusts (star ratings) is an important part of NHS policy. There are significant benefits to a Trust in being awarded the top rating of three stars. The methodology for awarding stars to acute hospitals is explained. The most significant areas of performance in determining a hospital's star rating are identified. The Department of Health claimed that performance in 2002 was better than in 2001, against standards that are getting tougher. This claim is assessed but no clear evidence is found to substantiate it. An alternative performance rating system using the same data is developed and compared with the official version to illustrate the importance of methodology--this system used only the outcome measures of hospital services included within the star ratings system, and not process measures. Only 41 per cent of hospitals would have received the same number of stars as their official rating. Some implications of the system of performance ratings are discussed. PMID- 14763104 TI - Securing the IT infrastructure. Protection can be solid and ongoing, even on a conservative budget. PMID- 14763105 TI - Emerging technologies. Tools considered fanciful just a few years ago have matured and are ready to break forth. PMID- 14763106 TI - U.S. health insurance forecast. Today's earnings are very strong but unsustainable. PMID- 14763107 TI - The Healthcare Collaborative Network. Its worth can be calculated in lives saved. PMID- 14763108 TI - On the heart beat. Needs are complex, and so are solutions. PMID- 14763109 TI - Enterprise change management. Today's healthcare enterprises need advanced technologies to manage the development environment. PMID- 14763110 TI - Toward a theology of sponsorship. PMID- 14763111 TI - Called to lead at Ascension Health. PMID- 14763112 TI - Building patient volume through "runway" management. PMID- 14763113 TI - Budgeting as theological reflection. True stewardship of resources requires realism, generosity of spirit, and commitment to the common good. PMID- 14763114 TI - Balancing act. Conscientious budgeting helps an organization carry its heritage into the future. PMID- 14763115 TI - Health care and the church's mission. An Australian bishop reflects upon the health care ministry's role in the larger work of the Catholic Church. PMID- 14763116 TI - Improving response to life-threatening illness. The nationwide CALL Care project involved 11 Catholic organizations. PMID- 14763117 TI - Making "caring connections". A new program enhances end-of-life care in a retirement community in Ohio. PMID- 14763118 TI - Ethical decisions in health care. A seven-step ethical discernment process can help organizational leaders make wise choices. PMID- 14763119 TI - Accountability and action. Affirming the five key elements of the board-medical staff relationship. PMID- 14763120 TI - A pathway to Franciscan excellence. A Louisiana health care system creates a process for developing leadership across the work force. PMID- 14763121 TI - To err is human; to report it is a quandary. PMID- 14763122 TI - Hasta la vista for Calif. nursing ratios? PMID- 14763123 TI - Mandatory compliance: never fun, sometimes inevitable. PMID- 14763124 TI - Dealing with death: HMGB1 as a novel target for cancer therapy. AB - High mobility group B1 (HMGB1) and its counter-receptor, receptor for advanced glycation end products (RAGE), represent suitable targets for investigation, integrating many aspects of modern biology, particularly that associated with chronic diseases involving inflammation, dysregulated cell death and cancer. Also known as amphoterin, HMGB1 was initially identified over 25 years ago as a transcriptional regulatory molecule causing DNA bending, and facilitating binding of several transcriptional complexes, in particular members of the nuclear hormone receptor family. Although loosely bound to chromatin, it is released from necrotic cells but not apoptotic cells and is actively secreted by activated macrophages in a partially tumor necrosis factor-dependent manner. It serves as a late mediator of septic death present within the serum and inflammatory sites of patients with arthritis, correlating with the inflammatory response, to signal tissue injury, causes sickness behavior, and acts as an endogenous pyrogen. Although known to interact with RAGE on endothelial cells causing activation and leukocyte recruitment, RAGE itself has most recently been shown to serve as a counter-receptor for leukocyte integrins, suggesting that signaling through this molecule is potentially important for cell adhesion and clustering as well as recruitment of inflammatory cells. Targeting the HMGB1 ligand or its receptor represents an important potential application in cancer therapeutics, given its widespread overexpression, as well as that of its receptor in virtually every tumor type carefully examined. This, coupled with its ability to accelerate tumor growth in immunodeficient murine models, suggests that it is a possible therapeutic target in patients with cancer. PMID- 14763125 TI - Strategies for treating cancers caused by multiple genome abnormalities: from concepts to cures? AB - The epidemiology, genetics/genomics and molecular biology of cancer all point to the involvement of a large number of genes in the malignant progression of the vast majority of human cancers. Our current conceptual models of cancer are discussed here and are integrated with an assessment of the strategies required for treating and potentially curing human cancers driven by multiple genome abnormalities. There are settings in which excellent responses will be seen in cancers driven primarily by single genomic abnormalities, e.g., imatinib in chronic myeloid leukemia and gastrointestinal stromal tumors. Other multigenic cancers will require drug cocktails or single drugs acting on multiple downstream targets. PMID- 14763126 TI - Multidrug resistance in cancer therapy: role of the microenvironment. AB - Despite advances in the design of chemotherapeutic agents, and although many of these effective agents are now available for clinical use, most human cancers are resistant to therapy at presentation or become resistant after an initial response. The effectiveness of chemotherapy is compromised by several microenvironmental factors that affect the bioavailability and efficacy of chemotherapeutic agents. These factors vary from one tumor to another, and from one location to another, within the same tumor. A better comprehension of microenvironmental multidrug resistance mechanisms would lead to a clearer understanding of the reason for chemotherapeutic failure. This improved knowledge will permit a more rational utilization of chemotherapy. PMID- 14763127 TI - HDAC inhibitors for the treatment of cancer. AB - Histone deacetylase (HDAC) inhibitors are a new class of cancer chemotherapeutics in clinical development that target the family of enzymes that catalyze the removal of the acetyl modification on lysine residues of proteins, including the core nucleosomal histones H2A, H2B, H3 and H4. The balance of nucleosomal histone acetylation is maintained through the opposing actions of histone acetyltransferases (HATs) and HDACs, and plays an important regulatory role in gene transcription. Alterations in both HATs and HDACs have been identified in tumor cells and may contribute to the altered gene expression found in many cancers. Inhibitors of HDAC activity induce cell cycle arrest, differentiation or apoptosis in tumor cells, and inhibit tumor growth in a variety of rodent models of cancer. Several structurally diverse HDAC inhibitors have entered clinical trials and are demonstrating encouraging antitumor activity in a variety of cancer types. As we learn more about these enzymes and the biological processes that they regulate, a strong rationale is emerging for the development of HDAC inhibitors as anticancer agents. PMID- 14763128 TI - Farnesyltransferase inhibitors as anticancer agents: current status. AB - Farnesyltransferase (FTase) is a major molecular target for the development of novel anticancer drugs that are designed to disrupt the aberrant signal transduction circuitry of tumor cells. The discovery more than a decade ago that farnesylation of Ras, one of the most prevalent oncoproteins in human cancers, is required for its cancer-causing activity was the initial stimulus to target FTase. However, it is now clear that yet to be identified farnesylated proteins other than Ras also play a pivotal role as targets for FTase inhibitors (FTIs). In this review, important mechanistic issues on the antiproliferative, pro apoptotic and anti-angiogenic activities of FTIs, with an emphasis on potential FTI targets, will be discussed. Important clinical issues associated with translational aspects of hypotheses-driven clinical trials will also be discussed. PMID- 14763129 TI - Raf pathway inhibitors in oncology. AB - Recognition of the importance of the Raf pathway in the proliferation and survival of tumor cells recently increased with the discovery of activating BRAF mutations in human tumors. Therefore, in addition to a role in controlling tumors with Ras mutations and activated growth factor receptors, inhibitors of the Raf pathway may harbor therapeutic potential in tumors carrying a BRAF oncogene. A variety of agents have been discovered that interfere with the Raf pathway, including antisense oligonucleotides and small molecules. These inhibitors block the expression of Raf protein, block Ras/Raf interaction, block its kinase activity, or block the kinase activity of the Raf target protein mitogen activated protein kinase kinase. Raf pathway inhibitors that are currently undergoing clinical evaluation show promising signs of anticancer efficacy with a very tolerable safety profile. Indeed, the Raf inhibitor BAY-43-9006 recently entered phase III clinical trials. Here, we review the current development status of potential Raf pathway therapeutics. PMID- 14763130 TI - Decitabine: development of a DNA methyltransferase inhibitor for hematological malignancies. AB - Decitabine (DAC) is a small molecule nucleotide analog that is incorporated into DNA and traps human DNA methyltransferases. Although initially developed as a cytotoxic agent, low-dose DAC is enjoying a revival as a specific inhibitor of hypermethylation in cancer. DAC has activity in several hematological diseases, especially myelodysplastic syndrome, chronic myelogenous leukemia and acute myeloid leukemia. Clinical and preclinical advances are presented in this review. PMID- 14763131 TI - ErbB family targeting. AB - Drugs for specific molecular targets have generated a great deal of excitement for their potential in cancer treatment, particularly with respect to our molecular understanding of cancer in recent years. The clinical utility of antibodies and small molecule kinase inhibitors has been demonstrated. The ErbB family of receptors is at the forefront of targets that are the subject of clinical trials. However, the activities of epidermal growth factor receptor antagonists have not been impressive as single agents. One of the lessons learned with this class of targets is that we currently do not know how to optimally apply them to the treatment of cancer. This review will discuss the issues contributing to this situation and the approaches that are currently being launched to resolve these issues. PMID- 14763132 TI - CKD-602. Chong Kun Dang. AB - Chong Kun Dang is developing the camptothecin analog CKD-602 for the potential treatment of cancer. By August 2000, phase II trials of CKD-602 were underway. PMID- 14763133 TI - CMT-3. CollaGenex. AB - CMT-3 is an orally active matrix metalloprotease inhibitor, and one of a series of inhibitors of multiple proteases and cytokines, under development by CollaGenex. The compound is currently undergoing phase II trials for the potential treatment of cancer, in particular metastatic cancer and HIV-related Kaposi's sarcoma. PMID- 14763134 TI - ZD-6474. AstraZeneca. AB - ZD-6474, one of a series of inhibitors of vascular endothelial growth factor receptor tyrosine kinase, which also has activity against the epidermal growth factor receptor tyrosine kinase, is under development by AstraZeneca for the potential treatment of solid tumors. Phase II trials in non-small-cell lung cancer, small-cell lung cancer and myeloma were ongoing in January 2003. PMID- 14763135 TI - MEN-10755. Menarini. AB - Menarini is developing the anthracycline aminodisaccharide MEN-10755 for the potential treatment of solid tumors. Six phase II trials began in 2000 to verify the therapeutic activity of the drug in a variety of tumors, including sarcoma and those of the ovary, breast, prostate and lung. PMID- 14763136 TI - Clofarabine. Bioenvision/ILEX. AB - Clofarabine is a purine nucleoside antimetabolite under development by Bioenvision (under license from the Southern Research Institute) and ILEX for the potential treatment of solid tumors, acute myelogenous leukemia, non-Hodgkin's lymphoma, and acute lymphoblastic and chronic lymphocytic leukemia. In September 2003, Bioenvision initiated a phase II trial in Europe in pediatric acute lymphoblastic leukemia, and in October 2003, ILEX submitted the first part of a rolling NDA to the FDA for the treatment of acute leukemia in children. PMID- 14763137 TI - [HER-ErbB family of receptors and their ligands: mechanisms of activation, signals and deregulation in cancer]. AB - The topic of this review is first to analyze in normal conditions the signal transduction pathways induced by members of the HER-ErbB receptor family and their ligands, and second, to decipher some deregulations occurring in various cancer types. As a result, new therapeutic opportunities will be mentioned. PMID- 14763138 TI - [Epidermal growth factor receptors: status evaluation methods and tumor expression]. AB - The development of targeted antitumor agent aiming the HER family of tyrosine kinase transmembrane receptors, focused on the issue of target detection. For HER2, the techniques used to detect an overexpression are well established and none has, to date, shown its superiority. These are immunohistochemistry (protein) and in situ fluorescent hybridization (DNA). For EGFR, the diversity of the activation means (amplification, mutation, enhanced transcription, ligands...) leads to technical caveats. Immunohistochemistry appears to be the most appropriate test for clinical use but standardized assays and scoring systems are mandatory. The EGFR levels of expression are very high in some normal tissues such as oral mucosae. In tumors, EGFR levels are variable. The highest expression is found in head and neck epithelial tumors. EGFR expression is also elevated in oral and lung dysplasia. EGFR testing may be required before targeted treatment. An exciting endpoint would be the functional and dynamic evaluation of EGFR and downstream proteins for patients, before and during treatment. PMID- 14763139 TI - [Prognostic value of epidermal growth factor receptor]. AB - The activation of the epidermal growth factor receptor (REGF) participates in oncogenesis by inducing cell proliferation, cell mobility and angiogenesis, and inhibiting apoptosis. This activation might be due to numerous abnormalities, including increased expression of its ligand. Although based on retrospective analyses with no standard technique of evaluation, the level of REGF expression is a prognosis factor for several tumors. It appears to be an indicator of poor prognosis which might influence treatments in head and neck tumors, cancers of the cervix, oesophagus, bladder and ovary. Its prognostic value is not observed in non-small cell lung cancer and remains to be demonstrated in adenocarcinomas such as colorectal tumors and beast cancer. Because of its role in oncogenesis and its prognostic value, REGF might in the future become a therapeutic target. PMID- 14763140 TI - [Pharmacological consequences of the targeting of epidermal growth factor receptor]. AB - Through clearly identified molecular mechanisms, EGFR targeting leads to cell physiology modifications as cell proliferation, apoptosis and DNA repair. These mechanisms can explain the proper activity of EGFR targeting drugs but also represent a strong rational basis on which several experimental studies have been designed with combinations of EGFR targeting and cytotoxic agents (chemotherapeutic drugs and radiotherapy). These data led to the development of clinical trials combining EGFR targeting agents and anticancer drugs. There is however a lack of experimental data concerning the importance of the drug sequences and above all the impact of EGFR tumoral expression on the effects of combinations. The lack of knowledge on resistance mechanisms to EGFR targeting is also a difficulty for an optimal development of EGFR targeting in the clinics. PMID- 14763141 TI - [Radiotherapy and inhibitors of epidermal growth factor receptor: preclinical findings and preliminary clinical trials]. AB - Recent studies have demonstrated that ionizing radiation activate existing cellular response pathways involving protein kinases. These pathways mediate the cytotoxic and cytoprotective responses of cell death and cell survival, respectively. Cytoprotective responses involve dominantly mitogen-activated protein kinase (MAPK) through radiation-induced activation of EGF receptors and may stimulate cell proliferation if radiation-induced damage is successfully repaired. Similarly, overexpression of EGF receptor family members or their activation by ligands expressed at normal levels may also confer radioresistance. Recent encouraging results indicate that EGF receptor inhibitors such as antibodies or small molecule tyrosine-kinase inhibitors may be effective radiosensitizers in tumors. Within the antibody class of EGF receptor inhibitors are monoclonal antibodies such as cetuximab and trastuzumab. These agents have a common target of the extracellular domain of the EGF receptor. Striking synergistic antitumor effects on human epidermoid and on adenocarcinoma cancer cell xenografts have been observed when cetuximab treatment is combined with radiotherapy. Promising results have also been obtained from the first clinical trial with cetuximab and radiotherapy in squamous-cell carcinoma of the head and neck. Trastuzumab has been poorly studied in combination with radiotherapy but showed an increased radiosensitivity of HER2-overexpressing breast cancer cells as measured by in vitro colony-forming assays. The mechanism of radiosensitization appears to involve DNA repair. There are well over a dozen agents in the small molecule tyrosine-kinase inhibitor category but the preclinical studies in combination with radiotherapy exist only for ZD1839 and CI1033. Preliminary studies confirm the capacity of ZD1839 and radiotherapy to produce a highly significant increase in tumor growth inhibition when compared to treatment with either modality alone. Another member of the quinazoline class of small molecule tyrosine-kinase inhibitors (CI1033) has recently been examined for its impact in conjunction with radiation in a series of HER-overexpressing breast cancer cell lines. This molecule inhibits tyrosine-kinase activity in all four members of the HER family, and preclinical studies showed a synergistic interaction of CI1033 with ionizing radiation. Finally, EGF receptor family member inhibitors may themselves be effective radiosensitizers and their use in future clinical investigations are based on a solid radiobiological rational. PMID- 14763142 TI - [Overview of epidermal growth factor inhibitors (with the exclusion of ZD1839 and C225)]. AB - The road of EGFR is an important road of initiation and progression disease. These receptors are highly expressed in the majority of tumours and this expression is associated with a poor prognostic, more aggressiveness, a poor response to treatment, and poor survival. Inhibition of EGFR is an interesting therapeutic approach. Out of ZD1839 and C225 many EGFR inhibitive agents are being evaluated in phase I, II and III trials. These agents that target the extracellular ligand-binding of the receptor include monoclonal antibodies (C225, EMD7200...) and complex ligand-toxins, others that target the intracellular ligand-binding of the receptor include small molecule tyrosine inhibitors (OS1774, ZD1839...). The results of phase I and II trials of the majority of these new agents are encouraging with a higher therapeutic index and lower toxicity than cytotoxic agents. It is necessary to confirm these results with phase II and III trials witch are now underway particularly for OS1774, EMD7200, ABX-EGF, PKI166, MD447 and ICR62. PMID- 14763143 TI - [Targeting of epidermal growth factor receptor and applications in ORL cancer]. AB - EGFR (epidermal growth factor receptor) is a transmembrane glycoprotein highly expressed in head and neck squamous cell carcinoma (HNSCC). Once triggered by ligands, tyrosine kinase located at their inner part is phosphorylated, initiating signal transduction pathways towards the nucleus. Two categories of EGFR inhibitors are affordable: the former group includes monoclonal antibodies whereas the latter regards tyrosine kinase inhibitors (ITK). Acting more as cytostatic than cytotoxic agents, they may potentiate both chemotherapy (CT) and radiation therapy (RT). Characterized by a spectrum of toxicity that does not overlap that of CT or RT, they may be associated with these treatments. First clinical trials have demonstrated the feasibility of their administration. Side effects merely consist of skin reactions and digestive symptoms; their intensity is generally mild and they resolve at the completion of treatment. As of yet, response rates are sometimes astounding but are still disparate. Randomized studies are ongoing. A better definition of EGFR status is warranted. Other data regarding interactions between her-family members, ligands parameters and the cascade regulation of signal transduction would certainly enable to better define the clinical applications of this new therapeutical approach. PMID- 14763144 TI - [Inhibitors of epidermal growth factor receptor and colorectal cancer]. AB - Colorectal cancers (CRC) express the epidermal growth factor receptor (EGF-R), a type I transmembrane receptor with tyrosine kinase activity. EGF-R signaling inhibition is a promising target for cancer therapy. ZD1839 (Iressa, AstraZeneca) and OSI-774 (Tarceva, Roche) are small molecular weight molecules with selective and reversible tyrosine kinase inhibition properties directed to EGF-R. Orally administered, these molecules induce sustained tumor stabilizations in previously treated metastatic CRC patients. The most frequent treatment-related toxicities are fatigue, diarrhea and acne-like follicular rash. The addition in the clinic of 5-FU, lOHP or CPT-11 to ZD1839 or OSI-774 does not seem to increase the own toxicity of each cytotoxic agents. Cetuximab (Erbitux, Merck) is an intravenously administered humanized monoclonal antibody which bind with high affinity with the extracellular domain of the EGF-R. The most frequent treatment-related toxicities are diarrhea, fatigue, nausea and cutaneous toxicity (allergic or acne-like follicular rashes, folliculitis). Most, if not all of these adverse events are mild. Partial responses were observed with cetuximab either alone (RR: 10%) or in combination with CPT-11 (RR: 22%) in patients with CPT-11 refractory advanced CRC which expressed EGF-R. The combination of cetuximab to folinic acid, 5-FU and CPT 11 seems tolerable at the cost of a slight increase of severe diarrhea and neutropenia. Finally, the promising activity of these EGF-R inhibitors has to be confirmed throughout randomized studies. PMID- 14763145 TI - [Therapeutic implications of epidermal growth factor receptor in lung cancer]. AB - Lung cancer is very frequent and associated with a high mortality. In the last 25 years therapeutic progress have been limited and do not allow a 5 year global survival rate exceeding to 13-14%. Tumor biology permits a better comprehension of cancerization mechanisms and offers hope of new treatments with targeted therapies which would be specific of cancer cells and so more efficient and less toxic. Epidermal growth factor (EGF) pathway and its receptor (EGFR) expressed by most lung cancer cells is the most successfully completed example. The bond of EGF with its receptor stimulates tyrosine kinase domain of EGFR and allows transduction of an activating signal. Inhibition of this signaling pathway stops tumor growth. Several agents are in development, from preclinical studies to phase III trials. It is a matter of humanized monoclonal antibodies, such as C225 (cetuximab), targeted against EGFR, or small molecules inhibiting tyrosine kinase activity of EGFR including ZD1839 (Iressa), OS1774 (Tarceva) or CI1033, and last antisense oligonucleotides. Antibodies and small molecules are well tolerated and are responsible for limited amount of side effects, mostly cutaneous toxicity and diarrhoea. Antitumor activity has been observed in monotherapy reaching up to 25% of clinical responses in the best series. EGFR inhibition seems to be also promising in combination with chemotherapy according to the synergy observed in preclinical studies and response rate up to 50% have been reported. But phase III studies have been disappointing and additional studies are warranted before consideration for a current daily practice, mostly that severe secondary effects were reported with pulmonary toxicities. In particular it remains to explain why clinical responses do not appear correlated with EGFR expression. PMID- 14763146 TI - [Expression of epidermal growth factor receptor and its ligands in other malignant tumor pathologies]. AB - The aim of this review was to describe expression of epithelial growth factor receptor (EGFR) and of its ligands in a panel of tumours (which are not specified elsewhere in this special issue of Bulletin du Cancer): squamous cell carcinomas, adenocarcinomas, sarcomas, brain and germ line tumours. The role of EGFR and its ligands in the carcinogenesis and the progression of these tumours, as well as the relevance of targeting EGFR. PMID- 14763147 TI - [Anti-epidermal growth factor receptor treatment: a new paradigm for conducting therapeutic trials]. AB - Agents which modify biological properties of tumour tissue can target many tenths of functions over- or underexpressed in human tumours. In general these agents are cytostatic rather than cytotoxic and will affect only that fraction of human tumours where the target plays and important and unique role for the viability of the tumour tissue. Alternatively it is expected that acute toxicity will not be observed at active dose-time exposure; rather subacute or chronic toxicity can be observed with these agents. Clinical studies will have to follow the following rules: characterisation of the pharmacological target and of its functional role on tumour tissue; definition of an optimal biological dose rather than a maximum tolerated dose; importance of validated pharmacodynamic endpoints; importance and thus need for early studies of combination regimens. It is still too early to define general guidelines for the study of these different therapeutic families. Nevertheless, studies already conducted with agents interfering with EGF mediated signalization have already permitted preliminary indications on pharmacodynamics, target assessment, level of activity and conduct of clinical trials with combination regimens. PMID- 14763148 TI - [Epidermal growth factor receptor and chemoprevention of epithelial tumor: rationale and development methods]. AB - Among the many pathways dysregulated during the carcinogenic process, REGF seems to be one of the most promising pathways to target in order to achieve chemopreventive and anticancer effects. Indeed, chemoprevention, the use of natural or synthetic compounds in order to reverse, suppress or prevent the carcinogenic process, aims at the cellular level at regulating the growth and sensitivity to apoptosis of premalignant and malignant clones. REGF activation leads to uncontrolled cellular proliferation, resistance to apoptosis, angiogenesis and invasion. Furthermore, REGF is frequently overexpressed in many epithelial tumors. This review will focus on the rationale and the ongoing research areas related to chemopreventive approaches targeting REGF. PMID- 14763149 TI - [Targeting epidermal growth factor receptor in cancer of the breast]. AB - The erbB receptor family is part of the receptor tyrosine kinase superfamily and consists of four members erbB. The erbB receptor family has been shown to play an important role in both the development of the normal breast and the pathogenesis and progression of breast cancer. Receptor overexpression has also been shown to be a negative prognostic indicator and to correlate with both tumor invasiveness and a lack of responsiveness to standard treatment, both chemotherapy and hormonotherapy. The targeting of EGFR mainly resides in two approaches: tyrosine kinase inhibition and monoclonal antibodies blocking ligand fixation. Many experimental data support the potential role of targeting EGFR in breast cancer. Particularly tyrosine kinase inhibitors demonstrates activity as single agent or in association with hormonotherapy, chemotherapy and trastuzumab. The association of Iressa with hormonotherapy points out that theses agents may prevent or differ hormonoresistance. Moreover studies in situ carcinoma suggest that tyrosine kinase inhibitors may play a role in chemoprevention. So, targeting EGFR may be indicated in a large spectrum of breast tumors from early to advanced stages, hormone negative or positive breast tumors. However the complexity of erbB network requires the targeting of multiple molecular sites within the network and the characterization of tumor profiles in order to optimally select patients for these therapies. PMID- 14763150 TI - [The potential for clinical guidelines to influence court decisions regarding the legal standard of medicine]. PMID- 14763151 TI - [New reversing agents that target membrane transporters]. PMID- 14763152 TI - [New method for analysis of target molecule in gastrointestinal carcinoma--from the choice of ATP7B as a target molecule for overcoming drug resistance through the development of an inhibitor against ATP7B]. PMID- 14763153 TI - [A case of esophageal achalasia with flat elevated lesion in the esophagus]. PMID- 14763154 TI - [A case of delayed traumatic diaphragmatic hernia presented with gastric volvulus 42 years after a blunt injury]. PMID- 14763155 TI - [A case of the patient with jejunal tube through the percutaneous endoscopic gastrostomy who vomited gastric juice frequently after the high fat enteral formula]. PMID- 14763156 TI - [Malignant lymphoma with submucosal invasion in the terminal ileum diagnosed with colonoscopy and examined by endoscopic ultrasonography]. PMID- 14763157 TI - [A long-term survival case of hepatic metastasis of rectal carcinoid in which etoposide was effective]. PMID- 14763158 TI - [A case of Tsutsugamushi disease complicated with cholecystitis]. PMID- 14763159 TI - Apoptosis regulators. AB - Over the last decade, a great deal of attention has been directed at elucidating the role of apoptosis regulators in governing survival decisions in neoplastic cells, particularly those of hematopoietic origin. A major focus of this work has involved investigation of the function of pro- and anti-apoptotic members of the BCL-2 family, and the relationship between these proteins and mitochondrial integrity. Currently, these proteins can be classified into two broad categories: those that modulate mitochondrial function and those that regulate the activation of caspases responsible for activation and execution of the apoptotic cascade. Within the first category, certain proteins (e.g., BCL-2, BCL-xL) act to preserve mitochondrial integrity by preventing loss of mitochondrial membrane potential and/or release of pro-apoptotic proteins such as cytochrome C into the cytosol. Other proapoptotic proteins (e.g., BAX, BAK, BIM) promote release of cytochrome C. These proteins are therefore primarily involved in regulation of the intrinsic, mitochondrial apoptotic pathway. Within the second category, proteins such as the inhibitors of apoptosis proteins (e.g., XIAP) or FLIP block the activation of caspases, particularly those involved in engagement of the receptor related, extrinsic apoptotic pathway. Cross-talk between the intrinsic and extrinsic pathways exists. For example, the BH3-domain only protein BID is cleaved by the activation of pro-caspase-8 through the extrinsic pathway, and translocates to the mitochondrion to promote cytochrome C release. Apoptosis is also regulated by various signal transduction pathways, possibly through post translational modifications in BCL-2 family proteins. For example, phosphorylation of BCL-2 through a JNK-dependent mechanism has been postulated to contribute to apoptosis induced by the taxane class of cytotoxic agents. Finally, attempts to modulate apoptotic pathways with small molecules have recently received much attention. For example, small molecule inhibitors of BCL-2 or mimetics of SMAC/DIABLO, which opposes the actions of XIAP, have recently been shown to promote the antineoplastic activity of conventional cytotoxic agents. It is likely that an improved understanding of apoptosis regulation will lead to new insights into neoplastic transformation, and may also provide important leads for the development of novel antileukemic strategies. PMID- 14763160 TI - Targeting aberrant transcriptional repression in acute myeloid leukemia. AB - Histone acetyltransferase (HAT) and histone deacetylase (HDAC) activities determine the acetylation status of histones, and have the ability to regulate gene expression through chromatin remodeling. Aberrant histone acetylation is known to play a key role in leukemogenesis. A common biologic feature, shared by genetically heterogeneous acute myeloid leukemias (AML), is a block of hematopoietic differentiation by the fusion proteins produced by chromosomal translocations. In many cases, a DNA binding fusion protein, which abnormally interacts with transcriptional co-regulators and increases local concentration of HDAC complexes, imposes a transcriptional repressive state on target gene promoters, which may become refractory to physiologic stimuli. To target this transcriptional repression, HDAC inhibitors (HDACI) have been developed, which are thought to derepress a set of genes whose transcriptional activation induces cell-cycle arrest, apoptosis and cellular differentiation and thus anti-tumoral activity. Therefore, HDACI might be utilized as effective antileukemic agents, and are currently under clinical trials. PMID- 14763161 TI - The mitogen-activated protein kinase signaling module as a therapeutic target in hematologic malignancies. AB - Important insights into the cellular and molecular biology of cancer in general and of hematologic malignancies in particular have been gained in the past two decades. The genes, and their protein products, involved in the transformation of cells from normal to neoplastic, as well as in the progression of neoplastic cells to a more aggressive, therapy-resistant phenotype, are being elucidated in great detail. However, most hematologic malignancies, particularly adult acute leukemias, remain associated with high mortality rates and new therapeutic approaches are urgently needed. The challenge is therefore to carefully and efficiently translate the information gained into effective therapeutic strategies. The clinical success achieved by tyrosine kinase inhibitors, such as ST1571 in chronic myelogenous leukemia, has stimulated interest for kinase-based signaling pathways as therapeutic targets in hematologic malignancies. The mitogen-activated protein kinase (MAPK) pathway is a common point of convergence of many different mitogenic and anti-apoptotic signal transduction pathways in hematopoietic, as well as epithelial, cancer cells and can now be clinically targeted by highly selective small molecule inhibitors. The mounting preclinical evidence of anti-leukemic activity of MAPK inhibitors, alone or in combination with pro-apoptotic small molecules or with conventional chemotherapeutic agents provides rationale that MAPK inhibition-based treatment strategies could soon enrich our therapeutic armamentarium against human leukemias. PMID- 14763162 TI - Targeting proteasome inhibition in hematologic malignancies. AB - Proteasome inhibitors represent potential novel anti-cancer therapy. These agents inhibit the degradation of multi-ubiquitinated target proteins mediating cell cycle progression, apoptosis, NF-kappa B activation, inflammation, cell cycle regulatory proteins such as cyclins and cyclin dependent kinase inhibitors, as well as immune surveillance; and regulate anti-apoptosis and cell cycle progression. Proteasome inhibitors also directly induce caspase-dependent apoptosis of tumor cells, despite the accumulation of p21 and p27 and irrespective of the p53 wild type or mutant status. Recent studies demonstrate that PS-341, peptide boronate, has remarkable anti-tumor activity in preclinical and clinical studies, not only in multiple myeloma but also in other malignancies. PMID- 14763163 TI - Targeting PI3K-AKT pathway for cancer therapy. AB - The phosphatidilinositol 3-kinase/protein kinase B (PI3K-AKT) pathway presents an exciting new target for molecular therapeutics. While exhibiting great promise, additional preclinical and clinical studies will be required to determine how best to target this pathway to improve patient outcome. A number of questions need to be answered prior to the implementation into patient care practices. As described below, the PI3K-AKT pathway regulates a broad spectrum of cellular processes, some of which are necessary to maintain normal physiological functions, which potentially contribute to the toxicity of the drugs targeting the pathway. Elucidation of the precise function of the PI3K-AKT isoforms, could promote the development of isoform specific approaches to provide a selective action on tumor cells. However, whether this will be possible due to conservation of structural domains is not yet clear. Inhibition of the PI3K-AKT pathway at multiple sites or a combination with inhibitors of different signaling pathways may allow the development of an acceptable therapeutic index for cancer management. Further, inhibition of the PI3K-AKT pathway combined with conventional chemotherapy or radiation therapy may provide a more effective strategy to improve patient outcome. As molecular therapeutics target the underlying defects in patient tumors, molecular diagnostics are required to identify patients with particular genetic aberrations in the pathway. It will be critical to provide adequate therapeutic strategies tailored to each patient. In addition, patients with different genetic backgrounds or in different health conditions could respond adversely to particular therapeutics. Therefore, identification of patients for particular drugs based on the underlying genetic defects in the tumor as well as the characteristics of the host would be of benefit for improving patient outcome. Linking the targeted therapeutics to molecular imaging approaches will determine appropriate biologically relevant dose for patients. It will also define expected tumor responsiveness and eventually will improve efficacy and decrease toxicity. In this regard, personalized molecular medicine is likely to soon provide effective cancer treatment. PMID- 14763164 TI - FIGO (International Federation of Gynecology and Obstetrics) annual report on the results of treatment in gynecological cancer. PMID- 14763167 TI - Carcinoma of the vulva. PMID- 14763168 TI - Carcinoma of the vagina. PMID- 14763169 TI - Carcinoma of the cervix uteri. PMID- 14763170 TI - Carcinoma of the corpus uteri. PMID- 14763171 TI - Carcinoma of the Fallopian tube. PMID- 14763172 TI - Carcinoma of the ovary. PMID- 14763173 TI - Gestational trophoblastic diseases. PMID- 14763174 TI - Gestational trophoblastic neoplasia, FIGO 2000 staging and classification. PMID- 14763175 TI - Cancer control in women. Update 2003. AB - The global cancer burden in women appeared to be increasing quickly at the end of the twentieth century with notable increases in the absolute numbers of cases of breast, cervix, lung and colorectal cancer of concern. However, prospects for cancer control in women appear to be good within our current knowledge and deserve close attention. Rates of lung cancer in women are increasing substantially in many countries and seem set to overtake breast cancer as the commonest form of cancer death in women in many parts of the world. These changes are due to the effects of cigarette smoking, a habit which women widely embraced during the second half of the last century. The high levels of smoking current in young women, which have yet to have their full inpact on death rates, constitute an important hazard not only for future cancer risks but for several other important causes of death. Although the breast is the commonest form of cancer in women in most western countries, the etiology of this disease remains elusive and preventable causes remain to be identified. Endogenous hormones also appear to have a role in cancer risk in women: oral contraceptives seem to increase slightly the risk of breast cancer in users in the use, and in the immediate post-use, period, but ten years after cessation the risk returns to that of never users. Oral contraceptive usage also appears to be protective against ovarian and endometrial cancer. The use of Hormonal Replacement Therapy (HRT) appears to increase the risk of endometrial cancer and a positive association with breast cancer risk appears to exist. Within the current knowledge of the epidemiology of cancer in women, the most important Cancer Control strategy is the prevention of cigarette smoking and the increase in the prevalence of adult women quitting smoking. Screening has also shown to be effective in reducing incidence and mortality of cervix cancer and mortality from breast and colorectal cancer. Although more work is needed, it is becoming clear that there could be an important role of HPV testing to further enhance cervix cancer screening. PMID- 14763176 TI - Promising directions for the diagnosis and management of gynecological cancers. AB - Diagnosis and management of cancer requires tools with both high sensitivity and specificity. The minimally invasive cervical smear has demonstrated how a test, even one with low specificity, can change the public health profile of a cancer from a late stage deadly disease to early diagnosis with rare tumor-related deaths. The benefit of such a test is best demonstrated by the low frequency of cervix cancer and its good outcome in countries where this test is readily available and used with appropriate secondary follow up. Early and specific symptoms, and identification and prevention for high risk groups has had similar impact for endometrial cancer. Neither a robust test, nor reliable or specific early symptoms are available for ovarian cancer, making clinical and scientific advances in this area a critical world-wide need. Current approaches testing one protein or gene marker at a time will not address this crisis expeditiously. New sensitive, specific, accurate, and reliable technologies that can be implemented using high throughput mechanisms are needed at as low a cost as possible. Ideally, these technologies should be focused on readily available patient resources, such as blood or urine, or as in the case of cervix cancer, minimally invasive informative approaches such as cervical smears. Techniques that allow data mining from a large input database overcome the slow advances of one protein one gene investigation, and further address the multi-faceted carcinogenesis process occurring even in germ line mutation-associated malignancy. Proteomics, the study of the cellular proteins and their activation states, has led the progress in biomarker development for ovarian and other cancers and is being applied to management assessment. Amenable to high throughput, internet interface, and representative of the proteome spectrum, proteomic technology is the newest and most promising direction for translational developments in gynecologic cancers. PMID- 14763177 TI - Quality of life in patients receiving treatment for gynecologic malignancies: special considerations for patient care. AB - Advances in the treatment of gynecologic cancer have extended the duration of survival of many patients. However, these patients frequently experience a variety of treatment- and disease-related side effects that diminish their quality of life (QOL) during and after treatment; among these are pain, nausea and vomiting, anemia, fatigue, peripheral neuropathy, emotional distress, and sexual dysfunction. Given the gains in survival time, patient care is being expanded to include enhancement or preservation of QOL in addition to early diagnosis and disease treatment, thus treating the whole person. In parallel with this evolution in cancer care, supportive measures are being increasingly recognized as crucial to effective patient management. This paper reviews some of the potential causes of diminished QOL in gynecologic cancer patients and basic treatment strategies for their control, with a focus on short-term QOL issues. It is important that clinicians monitor QOL during the course of the disease and its treatment, utilize procedures and therapeutic agents that take patient preferences and QOL into account, and proactively prevent and treat relevant symptoms. PMID- 14763179 TI - Quinacrine sterilization: reports on 40,252 cases. PMID- 14763180 TI - Quinacrine sterilization: a retrospective. AB - OBJECTIVE: To trace development of quinacrine sterilization (QS). METHODS: Review of published reports. RESULTS: The high prevalence of septic abortion among high parity women in Santiago, Chile, motivated Zipper to find a safe, inexpensive method of non-surgical female sterilization. Various cytotoxic drugs were tried in rats. Because quinacrine was already accepted for intrapleural injection it was chosen for the first clinical trial. A slurry consisting of quinacrine and xylocaine was instilled into the uterine cavity with a transcervical syringe. Reasonable efficacy was noted and a limited scar of the intramural tube demonstrated. However, a side effect of cortical excitation and reports of 3 deaths ended this approach. Zipper and Wheeler hypothesized that the difficulty was due to rapid absorption of quinacrine under pressure and designed a pellet form that dissolves slowly and could be delivered transcervically using a modified IUD inserter. A standard protocol of 252 mg in seven 36 mg pellets placed at the uterine fundus on two occasions a month apart has now been widely used with considerable evidence for safety and efficacy. Indeed, protection is greater than 98% at 2 years of use. CONCLUSION: QS is ready for widespread use, especially where surgical sterilization is not safely available or when women are poor candidates for surgery or have such a fear of surgery that they will not seek surgical sterilization. PMID- 14763181 TI - Quinacrine sterilization (QS): the ethical issues. AB - QS has generated debates that are ultimately grounded in various principles, norms, and values. Through a careful analysis of opposing arguments, this paper focuses on two ethical principles claimed by both sides, namely: respect for life and beneficence. Though issues surrounding QS are complex, from the common ground of these two principles, this paper proposes a course of action that addresses many of the concerns from both points of view. PMID- 14763182 TI - 25 years of quinacrine sterilization experience in Chile: review of 2,592 cases. AB - OBJECTIVES: To assess short-term side effects, long-term risks and efficacy of quinacrine sterilization (QS) in Chile. METHODS: Review experience of 2,592 cases sterilized with 2 or 3 transcervical insertions of 252 mg quinacrine as pellets since 1977; review the Chilean pre-clinical experience and epidemiological studies on cervical, endometrial and other cancers. RESULTS: Among 2,592 women who underwent QS, the total number of pregnancies was 119 (4.6%); 59 (49.5%) were carried to term with no birth defects related to QS. Nine cases were ectopic pregnancies. The ectopic pregnancy risk per 1,000 woman-years was 0.41, similar to that for surgical sterilization. The cumulative life-table pregnancy rates per 100 women at 10 years varied from 5.2 to 6.9. Mild and transient side effects were reported in 13.5% of quinacrine intrauterine insertions and pelvic inflammatory disease was diagnosed in 4 cases (0.15%). Long-term follow-up of quinacrine-sterilized patients shows no increased risk of cervical, endometrial or other cancer. CONCLUSIONS: QS efficacy at 10 years is comparable to widely accepted tubal clip and single point bipolar electrocoagulation laparoscopic procedures. QS has a low risk of serious, immediate side effects. No long-term risks have been identified after 25 years of use. PMID- 14763183 TI - Quinacrine nonsurgical female sterilization in Baroda, India: 23 years of follow up of 84 women. AB - OBJECTIVES: Evaluate the long-term effectiveness and safety of transcervical insertion of quinacrine hydrochloride pellets for nonsurgical female sterilization (QS). METHODS: During the period June 1979 through January 1980, 84 women were admitted to a study at the Baroda Medical College and Hospital, Baroda, India. Our protocol called for three transcervical insertions of 252 mg of quinacrine hydrochloride to be deposited in the uterus of each patient. Follow up was scheduled at 6, 12 and 48 months after the last administration. RESULTS: These women were 25 to 39 years of age at the time of the QS procedure and now, 23 years later, have completed their reproductive years. There were 4 pregnancies subsequent to the completion of QS, all prior to their 4-year follow-up. Thus, the life-time failure rate for these women was 3.7%. Complaints were minor, especially when compared to surgical sterilization. There were no long-term effects suspected of being attributable to QS. CONCLUSIONS: QS appears to be a reasonably effective method that is much safer than surgical sterilization. PMID- 14763184 TI - The rate of ectopic pregnancy for 24,589 quinacrine sterilization (QS) users compared to users of other methods and no method in 4 provinces in Vietnam, 1994 1996. AB - OBJECTIVE: To determine the rates of ectopic pregnancy with the use of quinacrine sterilization (QS) compared to other methods and no method (non-users). METHODS: Four provinces were selected for their above average numbers of women who had undergone QS: Nam Dinh, Nam Ha, Hai Duong and Hung Yen. Case histories related to surgical treatment of all ectopic pregnancies in these 4 provinces from 1994 through 1996 were collected from all hospitals by researchers from the Ministry of Health in June 1997. Using a questionnaire designed for this study, 120 physicians interviewed every woman in her home who had had an ectopic pregnancy during this period. If deceased, a family member was consulted. All interviews were completed in September 1998. The numbers of users of each method and nonusers were calculated from service statistics and demographic data. RESULTS: Based on 2,551,355 woman-years of exposure, the rate of ectopic pregnancy among users per 1000 woman-years was calculated to be: 0.26 with QS; 0.42 with surgical sterilization (TL) and IUD; 0.45 with the Pill; 0.50 with condoms; 0.78 relying on withdrawal; and 1.18 among non-users. CONCLUSION: Ectopic pregnancy rates for QS, TL, IUD and the Pill were similar and much lower than the rate for non-users of contraception. PMID- 14763185 TI - An FDA phase I clinical trial of quinacrine sterilization (QS). AB - OBJECTIVE: To review the significance of a United States Food and Drug Administration (FDA) approved Phase I clinical trial of a new use for an old drug, quinacrine. To discover whether ultrasound may have utility in quinacrine sterilization (QS). METHOD: This clinical trial began on 16 September 2000 at the Women's and Children's Hospital of Buffalo (WCHOB) in Buffalo, New York. Ten patients volunteered to have QS. These subjects were carefully followed with regularly scheduled examinations, including extensive laboratory blood tests. In addition, each patient had a trans-abdominal ultrasound examination six weeks or later past the date of the second insertion of quinacrine. The trial was completed on 30 April 2003. RESULTS: Laboratory results fell within normal limits, thus providing additional evidence to affirm the lack of toxic effects of QS. With ultrasound, we were able to see scars in both oviducts on all of our patients. One patient with a small scar as seen on ultrasound became pregnant. CONCLUSION: QS was found to be safe and effective. Ultrasound holds the promise of reducing the failure rate. PMID- 14763186 TI - A comparison of quinacrine sterilization (QS) and surgical sterilization (TL) in 600 women in Guizhou Province, China. AB - OBJECTIVES: Compare the safety and efficacy of quinacrine sterilization (QS) and surgical sterilization, also known as tubal ligation (TL). METHODS: 300 women accepted QS in Guiyang, China during the period from July 1995 to September 1997. Each patient was scheduled for follow-up at 3, 6, 12 and 24 months. In March 1998, a comparison group of 300 women electing TL during the same time period was systematically chosen. Researchers visited the village of every woman and conducted a structured interview. Each candidate was given a general health and pelvic exam at a clinic in her village. All interviews and exams were completed in August 1998. RESULTS: Of the 289 QS patients interviewed (a follow-up rate of 96.3%), 265 had had 2 insertions. There were 3 pregnancy failures for a cumulative life table failure rate of 1.2 per 100 women at 24 months. The 299 TL patients (a follow-up rate of 99.7%) had a similar rate of 0.7. There were no life-threatening side effects or deaths in either groups. QS was less disruptive, more easily tolerated, required fewer resources and was viewed more favorably than TL by women and their spouses. CONCLUSIONS: Both methods were found safe and very effective. However, QS was considered to be more acceptable than TL. PMID- 14763187 TI - Quinacrine female nonsurgical sterilization (QS): endometrial assessment by vaginal ultrasonography in 128 women. AB - OBJECTIVE: Investigate effectiveness, safety and endometrial pattern after QS. METHOD: This study began in March 1999 and ended March 18, 2003; 128 women received transcervical insertions of quinacrine. Follow-up visits with ultrasound were scheduled at 1, 3, 6, 12-month intervals. RESULTS: Two pregnancies occurred, one at 25 months, the other at 37. Adverse events (AE) were: yellow vaginal discharge, headache, mild abdominal pain, vaginal pruritus, nausea and transient decrease in endometrial thickness. One patient had allergic reaction. A third insertion was done in case of vaginal bleeding (16.4%). One year after QS 10% still had amenorrhea, which may be the result of the fact that 73% of our patients had received DMPH. Once inside the uterus, the dissolved quinacrine could be seen within seconds, via ultrasound as a "Lake of Quinacrine" which stays for up to two hours. Frequently, a transverse vaginal ultrasonographic view of the uterine cavity showed plug-like echogenic points at the cornua. CONCLUSION: Quinacrine sterilization is safe and effective. The echogenic points need to be more thoroughly studied in order to affirm whether ultrasonography may identify the blockage of the tubes. Since early pregnancy is due to imperfect tubal closure, the use of ultrasound may prevent failure. However, pregnancy due to later recanalization cannot be avoided. PMID- 14763188 TI - The effect of special training for quinacrine sterilization (QS) in Faisalabad, Pakistan: a report on an 1833-women subset of 11,000 cases. AB - OBJECTIVE: To determine the impact of retrained clinicians on the efficacy of transcervical quinacrine sterilization. METHOD: Retraining of clinicians in the accepted insertion technique was conducted in 1996. From 1 January 1997 through 2001, they performed 1089 quinacrine sterilizations in 11 MCH clinics of the Mother & Child Welfare Association at Faisalabad, Pakistan. Of these, 885 women had a recorded follow-up visit (81.4%) by 31 December 2001. RESULT: Reported pregnancy failures declined after retraining from 5.4% (SE 2.3) for one year of use to 1.1% (SE 0.4) for 5 years of use. The rates at 4 years of use showed the expected increase in failures for women 30 years old or younger of 1.5% (SE 0.7) compared to 0.9% (SE 0.4) for those over 30; a lower rate of 0.8% (SE 0.4) for breastfeeding subjects and 2.2% (SE 1.1) for mothers not breastfeeding; but rates were similar for uterine length and post insertion traumatic bleeding. CONCLUSION: Quinacrine sterilization appears safe with acceptable efficacy. PMID- 14763189 TI - Quinacrine sterilization (QS) among high-risk women: a study of 134 cases. AB - OBJECTIVE: To determine if quinacrine sterilization (QS) is safe and effective in women at high risk for surgery. METHODS: A trial was initiated at the Government Medical College in Patiala, India, in December 1993. Patient intake was terminated in July 1999 and the cut-off date for this analysis was March 31, 2003. Using a modified IUD inserter, seven 252 mg quinacrine pellets with 50 mg of diclofenac were transcervically inserted into the uterus. DMPA 150 mg was administered IM at the time of the first insertion as a back-up contraceptive. This same combination was inserted a month later. A total of 134 women underwent QS. Of these, 92 were considered to be at high risk for surgery, 27 were afraid of surgery or voluntarily opted for QS, and 15 had had failed surgical sterilization or surgery was found not to be technically feasible. Follow-up was scheduled for 1, 3, 6 and 12 months, and then annually after the second insertion or whenever side effects or complications were experienced. RESULTS: Mean follow up was 7.2 years. No pregnancies or serious complications were experienced. CONCLUSION: QS is a safe and effective option for women at high risk of surgical complications. PMID- 14763190 TI - The acceptability, efficacy and safety of quinacrine non-surgical sterilization (QS), tubectomy and vasectomy in 5 provinces in the Red River Delta, Vietnam: a follow-up of 15,190 cases. AB - OBJECTIVES: To compare the safety, efficacy and acceptability of quinacrine sterilization (QS), tubectomy and vasectomy in Vietnam. METHODS: This study was initiated in January 1998 and completed in February 2000. A sample of 9 districts in 5 provinces, where the prevalence of QS was known to be high, was selected. Every person sterilized in these 9 districts between January 1, 1988 and March 31, 1998 was identified and systematically interviewed by family planning clinicians who had received special training for this project. RESULTS: A total of 15,982 sterilization users were identified and 15,190 were interviewed and examined, including a gynecologic exam, if needed: a follow-up rate of 95%. Of those interviewed, 9,753 used tubectomy, 3,734 used QS and 1,703 used vasectomy. All three methods were found to be safe, although morbidity associated with tubectomy was more serious than with QS or vasectomy. No deaths were reported. After more than 5 years of follow-up, tubectomy had the lowest failure rate: 1.0%, followed by 4.1% with vasectomy. A pregnancy rate of 13.2% was reported with quinacrine, although only a small fraction of these failures were confirmed. A strong preference for QS was found. CONCLUSION: QS has an important role to play in sterilization services in Vietnam. PMID- 14763191 TI - A 22-year experience with quinacrine sterilization in a rural private clinic in Midnapore, India: a report on 5 protocols and 1838 cases. AB - OBJECTIVES: Evaluate the safety and effectiveness of quinacrine for non-surgical female sterilization in five different protocols. METHODS: The 5 trials were conducted sequentially. The first and largest, with 985 cases, tested the use of a curved inserter to place a 50 mg dose of quinacrine near each tubal ostia. The next 3 trials were carried out to determine the effect of adjunct procedures on the efficacy of the standard recommended protocol. The three adjuncts were 75 mg of intrauterine diclofenac, 10 mg medroxyprogesterone IM and either 10 mg of atropine IM or 20 mg of hyoscine butylbromide IM The final trial focused on the currently recommended protocol. RESULTS: The 100 mg dose placed at the tubal ostia with the curved inserter resulted in a failure rate of 9.0% at 20 years. Diclofenac or medroxyprogesterone did not improve efficacy over quinacrine alone. Atropine or hyoscine butylbromide substantially diminished the effectiveness of the quinacrine. The failure rate with the standard protocol in our series of 122 cases was 0.8% at 3.5 years. Side effects were minor. There were no deaths nor serious complications with any of these protocols. DISCUSSION: All 5 protocols appeared to be safe and the standard one was the most effective. PMID- 14763192 TI - Quinacrine sterilization (QS) in Iran and the use of HSG as a measure of success. AB - OBJECTIVES: To establish the safety, effectiveness and acceptability of quinacrine sterilization (QS) in Iran. To determine whether the hysterosalpingogram (HSG) performed under low pressure can be used to demonstrate success of the QS procedure rather than waiting for a pregnancy to occur in order to demonstrate failure. METHODS: This study was initiated in September 1990 in a private family planning clinic in Tehran, Iran. Patient intake for this analysis was completed 31 December 1998 and the cut-off date for follow-up data to be included in this analysis was 30 July 2002. During this period, 268 women received QS. From inception until April 1994, 160 women entered the study. The first 62 women received 3 insertions and the remainder received 2. Short-term side effects were closely followed in these 160 women. From 18 February 1994 until the patient intake cut-off date, 131 women entered the study and 46 of them received an HSG. RESULTS: With 4 to 12 years of follow-up there have been 7 pregnancies for a gross pregnancy rate of 2.6%. However, the use of the HSG tripled the risk of pregnancy for women who underwent the procedure. Furthermore, HSG, even when performed under minimal pressure, indicated failure of the QS procedure about 6% of the time when in fact both tubes would have closed had there been no intervention. Side effects were minor when compared to the complications of surgical sterilization. CONCLUSIONS: QS was found to be safe, effective and preferred over surgical sterilization by Iranian women. HSG understated the number of patients with bilateral tubal closure, or with tubes that would have closed given a little more time. PMID- 14763193 TI - Quinacrine sterilization of 1997 women in Daharpur, Midnapore, West Bengal, India: a comparison of 3 protocols. AB - OBJECTIVES: Determine the efficacy of two different dosage regimens of quinacrine placed at each cornual angle, employing a curved inserter, and for fundal placement of doses from 252 mg to 360 mg of quinacrine, depending on the age of the woman. METHODS: 1. The first trial involved 3 double insertions, a month apart, of 50 mg of quinacrine at each cornual angle. This trial was initiated on 14 August 1979 and completed on 26 June 1984 with 418 subjects admitted. 2. The second was a single double insertion of 100 mg at each cornual angle. This trial, initiated on 30 November 1984, was completed on 11 June 1985 with 100 subjects admitted. 3. The third trial began 2 January 1995, was completed 26 January 1998 and included 1479 subjects. There were 2 insertions, a month apart, with fundal placement of all pellets. The dose depended on the woman's age and ranged from 252 mg in the oldest to 360 mg in the youngest. The cut-off date for this trial was 23 January 2003. RESULTS: Only relatively minor side effects or complications were seen. None required hospitalization. Failure rates with multiple low dose or single dose cornual placement of pellets were unacceptably high. When higher doses of fundal placement of quinacrine were used at two visits, one month apart, no failures occurred. CONCLUSIONS: The third protocol shows great promise. PMID- 14763194 TI - Hysteroscopic and hysterosalpingographic study after intrauterine insertion of quinacrine pellets for non-surgical sterilization: results in 180 women. AB - OBJECTIVES: Document the effects on the tube and uterus of one, two and three doses of 252 mg of quinacrine. METHOD: The study included 180 fertile women seeking permanent contraception at the Shatby Family Planning Clinic in Alexandria, Egypt, in 1988. All cases received three applications of seven 36 mg quinacrine hydrochloride pellets during the proliferative phase of three consecutive menstrual cycles. The patients were randomly divided into groups A, B and C. Hysterosalpingography (HSG) was performed on the 6th day of menstruation and hysteroscopy on the 10th day of the same cycle after the first application in group A, the second, in group B and the third, in group C. The study was concluded in 1999. RESULTS: HSG showed 52 cases of bilateral obstruction, four of bilateral patency, and four of unilateral patency in group A. All in groups B and C elicited bilateral tubal obstruction. Cornual obstruction was seen in 33%, 65% and 85% in group A, B and C, respectively. Intramural obstruction was found in 50%, 33% and 10% in the three groups. Isthmic tubal obstruction was detected in 8%, 2.5% and 5% in groups A, B and C, respectively. Four types of ostial appearances could be recognized hysteroscopically. Type 0 (patent tubes), Type I (distal tubal blockage), Type II (intramural obstruction) and Type III (cornual obstruction). In group A, Type 0 was evident in 10%, Type I in 8%, Type II in 50% and Type III in 33% of cases. The respective figures in group B were 0%, 2.5%, 33% and 65%, while in group C, they were 0%, 5%, 10% and 85%. Hysteroscopy showed no abnormal endometrial findings in group A, but 35% and 85% of cases in group B and C showed some changes. CONCLUSIONS: Two applications of quinacrine were 100% effective. The side effects of quinacrine pellet applications were minimal and well tolerated by all the users. The possibility of reversal of the procedure is outlined. PMID- 14763195 TI - Female sterilization with quinacrine using hysterosalpingography (HSG) as an endpoint after a single-insertion protocol in Caracas, Venezuela. AB - OBJECTIVE: To evaluate the intrauterine insertion of quinacrine as an alternative nonsurgical female sterilization method by confirming bilateral occlusion of the fallopian tubes using HSG in a group of women who desire permanent sterilization. METHODS: After doing hysterosalpingography to confirm patency of both fallopian tubes, 324 mg of quinacrine were introduced with a modified IUD inserter in 30 patients who came to Concepcion Palacios Maternity Hospital seeking permanent sterilization, between June 2000 and September 2001. Follow-up with HSG was done 3 months later to verify occlusion of the fallopian tubes. RESULTS: 26 of 30 patients (86%) had bilateral tubal occlusion as determined by HSG. There were minor side effects such as: pain (66.7%), yellow discharge (100%) and menstrual abnormalities (13.3%). One woman became pregnant after HSG showed bilateral occlusion. HSG may interfere with the action of the quinacrine. CONCLUSION: QS is a simple and safe alternative to surgical sterilization with few side effects. PMID- 14763197 TI - Quinacrine sterilization (QS) in a private practice in Daytona Beach, Florida: a preliminary report. AB - OBJECTIVES: To affirm that QS can be performed safely and effectively in a U.S. private office practice. METHODS: The U.S. FDA Modernization Act of 1997 Pharmacy Compounding Provisions made it possible for American physicians to begin offering QS to their patients. These provisions became effective November 21, 1998. This series was initiated in October 2000. The standard protocol recommended by the International Federation for Family Health (IFFH) is followed. Information on patients is recorded on forms suggested by IFFH to accomplish good post-marketing surveillance. The potential role of uterine septae in QS failures is of particular interest to this investigator. RESULTS: Seven cases have been completed. There have been no failures. Side effects have been minor. Women have been exceptionally happy with this method. The Florida Agency for Health Care Administration has examined QS and found it to be an acceptable off-label use of quinacrine. CONCLUSION: Preliminary results have been similar to those reported by QS researchers around the world. PMID- 14763196 TI - Quinacrine sterilization in Libya: 200 cases. AB - OBJECTIVE: Document the safety, effectiveness and acceptability of quinacrine sterilization (QS) as an alternative to surgical sterilization in Libyan women. METHODS: This study was initiated 1 October 1998 at the Misurata Central Hospital and Lamis Clinic. Patient intake was completed 30 September 2002. The cut-off date for this analysis was 31 December 2002. A total of 200 women were given 2 doses, each consisting of 252 mg of quinacrine hydrochloride in the form of 7 pellets inserted one month apart. They were placed at the uterine fundus during the proliferative phase of the menstrual cycle using a modified IUD inserter. Women were asked to report any unusual observations or side effects and instructed to use a barrier method or safe period for one month from the time of the first insertion. Follow-up was scheduled at 3, 6 and 12 months after the date of the second insertion and every 6 months thereafter. RESULTS: Sixty-six women have been monitored for up to 3 years and follow-up of all patients continues. There has been no loss to follow-up. No side effects of any consequence have been reported. Thus far, no pregnancies have been reported for this protocol. CONCLUSIONS: Findings in this study are consistent with those seen in other countries. QS has been shown to be safe, effective and acceptable among Libyan women. PMID- 14763198 TI - Quinacrine sterilization (QS) experience in the Philippines: a preliminary report. AB - OBJECTIVE: The first clinical trial of Quinacrine Sterilization (QS) in the Philippines was undertaken in Cebu City on January 10, 2000, to evaluate the acceptability, safety, effectiveness and side effects of this technology. We intend to recruit 500 patients to utilize this technique for limiting family size. For the purposes of this report, our cut-off date is April 11, 2003. METHODS: Over more than two years, QS was performed on 36 volunteer patients. After careful explanation of the procedure and given the opportunity to ask questions, they had signed an informed consent. The trial involved transcervical insertion of 252 mg quinacrine in the form of pellets, and placed at the tip of the uterine fundus on two occasions, a month apart. Condoms were routinely provided to all patients except those on oral contraceptive pills and DMPA after the first insertion to be used for six weeks after the second one. As the numbers are small, no statistical evaluation was called for. RESULTS: The accumulated experience was 515 woman-months. There were no pregnancies, neither ectopic nor intrauterine: Adverse events (AE) were mild. Some patients complained of a yellow discharge and itching. Fifty percent experienced mild abdominal discomfort which was easily managed with mefenamic acid. CONCLUSIONS: Although this is a small study, we believe that QS is both safe and effective and we are strongly encouraged to continue to offer this nonsurgical sterilization method to our patients. PMID- 14763200 TI - 8-year follow-up in a randomized trial of one vs two transcervical insertions of quinacrine pellets for sterilization in Indonesia. AB - OBJECTIVE: To evaluate the efficacy of one vs two insertions of quinacrine and the long-term safety of quinacrine sterilization (QS) 8 years after the procedure in Indonesia. METHODS: Between March 1993 and September 1995, a randomized trial was conducted in 6 academic centers in Indonesia. In February 2003, a follow-up study was undertaken in Bandung, one of those centers. This survey required a home visit of each woman. A questionnaire was designed to elicit information regarding current general health status, method failure, pregnancy outcomes and other contraceptive methods now used by women who experienced failures. Among the 70 patients receiving a single insertion of quinacrine pellets, 14.3% had become pregnant. There were no pregnancies among the 30 who received 2 insertions. All the women were found to be in good health. No long-term side effects or complications were identified. CONCLUSION: The two-insertion protocol is unmistakably superior to the single insertion. This study provides further evidence that QS is a safe contraceptive method. PMID- 14763199 TI - Marie Stopes Society, Pakistan: 1000 cases of quinacrine sterilization (QS). AB - OBJECTIVES: To evaluate the safety, efficacy and acceptability of QS in Karachi, Pakistan. METHODS: 1000 women who had chosen sterilization during the 4-year period 1994 to 1997 inclusive were offered QS at both stationary clinics and in a mobile van at 23 sites in the outskirts of Karachi. The protocol involved transcervical insertion to the uterine fundus of 252 mg quinacrine in 7 pellets and 55 mg of ibuprofen in 3 pellets through an IUD inserter, during the proliferative phase of the menstrual cycle. Two doses were administered one month apart. A temporary method of contraception was provided for 3 cycles, usually DMPA. Follow-up was scheduled: monthly for 3 months, quarterly for 1 year and then every 6 months for 4 years. RESULTS: The crude pregnancy rate after 4 years was 2.0%. Minor complications and complaints were reported by 59% of the patients. There was one ectopic pregnancy and no major complications. CONCLUSIONS: QS was found to be safe and effective and has become the most popular method of sterilization in our area of Pakistan. PMID- 14763201 TI - Quinacrine sterilization (QS) in Syria: a preliminary report on 297 cases. AB - OBJECTIVES: To evaluate the safety, efficacy and acceptability of quinacrine sterilization (QS) in Syria. METHODS: From July 2001 to December 2002, 297 women who requested permanent sterilization volunteered for QS either in my private practice or my local family planning center in Aleppo, Syria. The standard protocol was used: 252 mg of quinacrine in the form of 7 pellets are deposited at the uterine fundus with a modified CuT IUD inserter during the proliferative phase of the menstrual cycle. This procedure is repeated 4 weeks later. DMPA was injected at the time of the first insertion for temporary contraception. Every sterilized woman has had a monthly checkup visit until the cut-off date for this report, including a beta HCG pregnancy test. All procedures were performed by the author. The cut-off date for this report was June 11, 2003. RESULTS: The single pregnancy was ectopic. Four women (1.3%) complained of severe pain. Moderate pain was experienced by 13.1% while the remaining women felt mild pain, all easily treated. The remaining side effects were minor and also easily treated. Oligomenorrhea and amenorrhea affected 29% of the women and lasted for several months. Immediate side effects are similar to reports from other researchers. CONCLUSIONS: Results thus far regarding efficacy are encouraging. QS has proven to be acceptable. PMID- 14763202 TI - 10-year follow-up of women who elected quinacrine sterilization (QS) in Wonosobo, Central Java, Indonesia. AB - OBJECTIVES: To evaluate the safety and efficacy of quinacrine sterilization (QS) in Indonesia. METHODS: During the period, August 1992 to October 1993, 200 women who had requested surgical sterilization volunteered for QS at the Wonosobo Regency Hospital, Central Java Province, Indonesia. The protocol called for transcervical insertion of 252 mg of quinacrine in the form of 7 cylindrical pellets and 55.5 mg of ibuprofen with a CuT-IUD (Kimia Farma) inserter during the proliferative phase of the menstrual cycle. A second procedure was done 4 weeks later. The technique used is essentially the same as inserting a CuT-IUD. Follow up was scheduled at 6, 12, 24 and 48 months after the last insertion. In March 2003 additional monitoring was completed. RESULTS: The 10-year cumulative pregnancy rate was 4.3 per 100 women with a follow-up rate of 93%. No pregnancies had occurred among these women since the 4-year follow-up. No long-term side effects or complications were reported. CONCLUSIONS: After 10 years of use, QS was found to be safe and reasonably effective. PMID- 14763203 TI - Quinacrine sterilization (QS) in Costa Rica: 694 cases. AB - OBJECTIVES: To evaluate the safety, efficacy and acceptability of quinacrine sterilization (QS) in Costa Rica. METHODS: From 1989 though August 1993, 694 women volunteered for QS in my private practice. All were referred by a family planning clinic or a local hospital obstetric service. The protocol used involved the transcervical insertion of 216 mg of quinacrine hydrochloride in the form of 6 pellets. A second dose was given 4 weeks later. All insertions were done in the first 14 days of the menstrual cycle. The procedure was similar to the CuT IUD placement. Temporary contraception was recommended for 3 months after the last insertion. The cut-off date for this analysis was April 1994. RESULTS: With 7 months to 5 years of follow-up, the gross pregnancy rate was 2.5%. Side effects were relatively minor, none requiring hospitalization. CONCLUSION: QS was found to be safe, effective and acceptable. PMID- 14763204 TI - Quinacrine sterilization (QS): informed consent. AB - Informed consent is a basic human right for any medical procedure. It is particularly important that women know what is involved in any sterilization method, and how it will affect their health and their emotional life. Over 140,000 QS procedures have been performed in 34 countries. In no country has there been any formal effort to advance the acceptance of this method. Instead, satisfied users have been the promoters. Thoroughly informed consent is vital to patient satisfaction. A working group undertook an initiative to create an ideal consent form. The product of that initiative is presented. PMID- 14763205 TI - Treating distressed couples with coexisting mental and physical disorders: directions for clinical training and practice. AB - Relationship difficulties frequently coexist with significant emotional, behavioral, and physical health problems in one or both partners. Moreover, couple therapists report individual emotional and behavioral disorders as among the most difficult problems that they encounter in clinical practice. In this article, we summarize empirical evidence regarding the association between relationship distress and the onset, course, and treatment of mental and physical health problems. Based on these findings, we propose five critical implications for clinical training and the practice of couple therapy. PMID- 14763206 TI - Close relationships research: a resource for couple and family therapists. AB - This article describes the relatively new field of close relationships research, offering a representative list of topics studied by relationship researchers. Some of the common interests shared by both close relationships researchers and couple and family therapists are described, with the shared emphasis on relationships as an anchor for both fields. Some representative love theories are discussed, and Love Styles theory and research are presented in considerable detail. A clinical case example indicates how love styles research may be employed to advantage by couple therapists, and the utility of other close relationships theories and measures for therapy is briefly discussed. PMID- 14763207 TI - The Intimate Justice Scale: an instrument to screen for psychological abuse and physical violence in clinical practice. AB - This article describes development of the Intimate Justice Scale (IJS) and reports on a clinical study of the validity, reliability, and clinical usefulness of the instrument. Rather than measuring specific acts of abuse, the IJS measures ethical dynamics of couple relationships, which are evident in patterns of action and attitude expressed over the course of the relationship. Ethical dynamics appear to correlate with partner abuse. The study suggests that the IJS may reliably identify victims of abuse and may discriminate between minor and severe levels of abuse. The IJS can be completed and scored in less than 10 min and may be useful for screening in mental health, medical, and social service agencies. Clinical guidelines and a case example are presented. PMID- 14763208 TI - Witnessing the effects of political violence in families: mechanisms of intergenerational transmission and clinical interventions. AB - In this era of globalization, when news about political violence can haunt anyone, anywhere, those whose families have suffered political violence in the past are particularly vulnerable to current distress. Skilled in understanding transgenerational processes, family therapists need to be familiar with the mechanisms by which children are exposed to the effects of political violence suffered by their elders-that is, the ways in which they become their witnesses. This article presents a framework for understanding how the trauma of political violence experienced in one generation can "pass" to another that did not directly experience it, and proposes a model to guide clinical intervention. PMID- 14763209 TI - Trends in family therapy supervision: the past 25 years and into the future. AB - This investigation repeats a survey of American Association for Marriage and Family Therapy (AAMFT) Approved Supervisors done in 1976 and 1986. Despite negative contemporary contextual influences, the AAMFT Approved Supervisor credential is still attractive to the same percentage of the AAMFT clinical membership as formerly and reflects in general the demographic characteristics of marriage and family therapists (MFTs) in clinical practice. Across the last quarter century there has been a steady increase in female AAMFT Approved Supervisors and those with terminal master's degrees. These and other trends are discussed with a view toward what supervision is becoming and what it needs to become. PMID- 14763210 TI - A place for marriage and family services in employee assistance programs (EAPs): a survey of EAP client problems and needs. AB - Marriage and family services have not been widely recognized as part of employee assistance programs (EAP), although family and relational problems are widely cited as sources of problems on the job. EAP clients (N = 800, 97% self-referred) indicated how much family, psychological/emotional, drug, alcohol, employment related, legal, and medical problems troubled them and the need for services in each area. Psychological/emotional (66%) and family (65%) problem areas frequently were rated "considerable" or "extreme." Both areas were rated as "considerable" or "extreme" by 48.6% of participants. In view of the evidence that marriage and family services can be effective with both family and psychological/emotional problems, professionals who are competent to provide such services have much to offer EAP programs. PMID- 14763211 TI - Talking about race using critical race theory: recent trends in the Journal of Marital and Family Therapy. AB - This study uses critical race theory as an interpretive lens to critique recent race related articles in the Journal of Marital and Family Therapy (JMFT). Our primary goal is to contribute to and inspire dialogue about the perspectives marriage and family therapists (MFTs) are taking in relationship to race. We situate our exploration within the broader context of continuing professional education. We describe the main themes of critical race theory and use them as the conceptual framework. Analyzing 127 articles, we found that only topics related to couples and divorce occurred more frequently than race and social justice. Within the articles on race, evidence suggests that issues of race and racism are emerging as key informants of MFT practice. We point to areas for consideration in future MFT research and practice. PMID- 14763212 TI - Breaking down the barriers to clinical service delivery: walk-in family therapy. AB - This article provides a description of a unique walk-in family therapy service designed to overcome barriers to clinical delivery, as well as a survey of client experiences with the therapy. This therapy modality is aimed at providing an immediately accessible, affordable, nonstigmatizing, single-session-focused resource. The service operates from a systemic, collaborative, consumer oriented, pragmatic, non-pathology perspective. Follow-up telephone interviews were conducted with 43 clients 3 to 6 months after they received treatment. Generally, former clients reported satisfaction with the service. The majority (67%) indicated some level of improvement, and 43% of participants found their single session sufficient to address their concerns. At the time of the session, the majority (86%) of clients were rated by their therapists as "customers" regarding their motivational readiness for change. PMID- 14763213 TI - On the 100th birthday of Prof. Dr. med. Juan Manuel Tato: pioneer in advancing otorhinolaryngology into modern neurootology. PMID- 14763214 TI - Computerized ultrasonographic craniocorpography and abnormal psychomotor activity in psychiatric patients. AB - A new version of craniocorpography (CCG), called computerized ultrasonographic CCG (Comp-USCCG), has been clinically applied for objective recording, documentation, and quantitative evaluation of abnormal psychomotor activity in psychiatric patients. Implications of this completely new approach to psychopathology are discussed. An original representation of Comp-USCCG data (introducing the time dimension as a new CCG parameter) is used to illustrate better the atypical abnormal stepping Comp-USCCG movement patterns in psychotic patients, some of which have not been described in neurootological patients to date. These atypical abnormal stepping Comp-USCCG movement patterns are prolonged longitudinal or shortened or backward longitudinal displacement; dysrhythmic longitudinal or lateral sway; and longitudinal or lateral directional changes. Reflecting the abnormal psychomotor activity, Comp-USCCG also provides for possible indirect evaluation of the underlying subjective psychotic experience. The contribution of the approach could be defined as an application of a known neurootological method into a new field of medicine (psychiatry) with a new purpose (to record and measure abnormal psychomotor activity). Our conclusion is that Comp-USCCG could become the first objective and quantitative method available for use in the field of clinical psychiatry. PMID- 14763215 TI - Quantitative electroencephalography: preliminary report--tinnitus. AB - This preliminary report is an account of 21 consecutive patients who had tinnitus of the severe disabling type and were examined with quantitative electroencephalography (QEEG). A multimetric analysis of the raw data was highlighted by an abnormal incidence of significant central nervous system electrical dysfunction identified in each patient (21 of 21). Relative power was increased or decreased in the temporal region in 10 of 21 patients. Relative power was reported to be increased or decreased in temporal frontal regions in 20 of 21 patients. Coherence irregularity was identified in all 21 patients. QEEG preliminary data support the hypothesis of a final common pathway for tinnitus and the significant role of the temporal and temporofrontal regions of interest in patients with tinnitus of the severe disabling type. QEEG with multimetric analysis is considered a significant addition to the medical audiological tinnitus patient protocol as one of a battery of electrophysiological tests for the clinical identification of a predominantly central type of tinnitus. PMID- 14763216 TI - Neurofeedback and quantitative electroencephalography. AB - This study was conducted in an attempt to determine the efficacy of neurofeedback (NFB) in the treatment of patients suffering from vertigo or tinnitus. Results indicated that after NFB, power for delta and theta bands was reduced; however, an increase of power was noted for the alpha bands. Furthermore, normalization was observed for the vestibular evoked potentials (VestEP). After NFB, a normalization of the VestEP was also demonstrated in a patient suffering from a bilateral tinnitus. A follow-up study (12 months after NFB) demonstrated that the VestEP were normal. PMID- 14763217 TI - Effects of noise on the intensity of distortion product otoacoustic emissions. AB - Distortion product otoacoustic emission (DPOAE) is a widely used differential diagnostic method for diagnosing inner ear disorders. It provides us information on the condition of the outer hearing cells (OHCs). Otoacoustic emission occurs only when the OHCs are functioning normally. Changes in thresholds of DPOAE curves can provide us important information on the activity of the OHCs. The inner ear shows nonlinear properties if the OHCs are functioning normally. If OHCs are injured and thus function improperly, the system stops showing nonlinear properties. If we have a system with periodic excitation and, with the addition of white noise, the signal-to-noise ratio on the output increases (at least for small noise intensities), we have witnessed what is called phenomenon stochastic resonance. Our goal was to elucidate how white noise influences the intensity of DPOAE. If there is emission, that specific ear surely exhibits nonlinear behavior, which in turn is the basic property needed for stochastic resonance. PMID- 14763218 TI - Computerized analysis of established craniocorpography. AB - This study reports 21 patients who suffered from vertigo and underwent a complete otorhinolaryngological evaluation and otoneurological tests to establish a peripheral labyrinthine disorder. All patients were given the Unterberger stepping test in a motion analysis laboratory. Their results were compared to those of a population of 21 normal individuals who were analyzed in the same motion laboratory. A precise description of sway, angular deviation, and test duration was obtained from the VICON motion analysis program. As expected, angular deviation was statistically greater in the group with vestibular disorders. PMID- 14763219 TI - Evaluation of oculomotor tests in patients with tinnitus. AB - Tinnitus as a symptom remains a serious multidisciplinary problem. Vertigo or dizziness is not noticed frequently in tinnitus patients, so vestibular function is not often studied. Because they are in close proximity to one another, the vestibular and hearing organs may influence each other. We decided to evaluate the results of the oculomotor reflex in tinnitus patients. We carried out clinical examinations and audiological and oculomotor tests in 50 tinnitus patients and 30 healthy persons. Such oculomotor tests as smooth-pursuit eye movement, optokinetic tests, and saccadic eye movement and gaze fixation were performed. Tinnitus was unilateral in 37 tinnitus patients (74%) and bilateral in 13 (26%). Twenty-two tinnitus patients (44%) reported previous remote episodes of vertigo and dizziness. Gaze-evoked nystagmus was absent in all cases. Morphological abnormalities and gain decreases in smooth-pursuit tests were recorded in 19 tinnitus patients (38%). Findings in the optokinetic test were incorrect in 10 (20%). The saccadic eye movement test showed disturbances in 18 tinnitus patients (36%). In 11 (22%), abnormal recordings were found in two tests and, in 10 (20%), abnormal recordings were found in all three tests. Our study suggests that computerized, quantitative electronystagmographic analysis in tinnitus patients should be interpreted carefully. A persistence of abnormal oculomotor recordings can suggest subclinical central vestibular system impairment. PMID- 14763220 TI - Cognitive disorders: diagnosis and treatment. AB - During the last half century, medical developments have expedited an increase of life expectancy all over the world. Owing to this extension of life, some pathological processes that formerly were considered to exhibit low prevalence now have become top priorities for public health programs. Among these pathological processes, cognitive diseases stand out, the two most frequent being Alzheimer's disease and cognitive disorders of vascular etiology, for which humans are at increased risk beyond age 65. We studied 120 adults with cognitive disorders representing a variety of etiologies. We performed a comparative statistical study between members of this group of patients and a group of general patients by means of different neurophysiological examinations: acoustically evoked potentials, P300, brain electrical activity mapping both in the resting state and under complex stimulation, and Doppler ultrasonographic extracranial and transcranial assessment. The comparative study between both populations revealed significant differences in results of the various tests. Neurotransmitter equilibrium (dopamine, acetylcholine, gamma-aminobutyric acid [GABA], etc.) in the areas of the cerebral cortex related to cognitive disorders is essential in the diagnosis and treatment of these diseases. PMID- 14763221 TI - How does cochlear implantation affect the contralateral vestibular system? AB - Cochlear implantation has been performed for 16 years by investigators at Semmelweis University. During this period, different types of cochlear implants have been used and, in 30% of cases, hearing was observed to be restored in the nonimplanted ear. In addition to contralateral hearing improvement, significant improvement was observed in the caloric responsiveness of the nonoperated labyrinth. The preoperative median value of the average slow-phase velocity of the caloric test increased, and the increase was statistically significant on the contralateral side. The reason for this caloric response improvement is unclear, although possible explanations are brain plasticity or presently obscure trophic influence on the vestibular system. Whereas the role of brainstem function in the improvement of the contralateral ear's caloric response remains unclear it is also possible that hearing impulses affect the labyrinth. Clearly, the influence of cochlear implants on vestibular function requires further investigation to explain the improvement of contralateral vestibular responsiveness. PMID- 14763222 TI - Intratympanic dexamethasone treatment for control of subjective idiopathic tinnitus: our clinical experience. AB - In this report, we summarize our clinical experience with intratympanic dexamethasone treatment (IDT) for control of tinnitus. From March 2000 through February 2001, we observed 54 patients (23 women, 31 men; mean age, 49.6 +/- 7.2 years; range, 24-71 years) suffering from subjective idiopathic tinnitus (SIT). After common audiological tests had been performed; all patients underwent specific topodiagnostic tests to verify the cochlear SIT genesis. The 50 subjects with positive results from a furosemide test and negative results from caraverine and carbamazepine tests were selected for the IDT, consisting of transtympanic perfusion of 4 mg dexamethasone to the round window via the middle ear. The treatment was repeated three times daily for 3 consecutive months. Its short-term effects were evaluated 2 weeks after the last perfusion. In 17 of 50 of these patients (34%), the SIT disappeared; 20 of the 50 (40%) reported a significant decrease of the symptom; and the remaining 13 of the 50 (26%) did not experience any improvement. Therefore, we believe that IDT represents an effective drug delivery system for SIT control, as long as the condition arises from inner ear disorders only and treatment occurs within 3 months of symptom onset. PMID- 14763223 TI - Fixed combination of cinnarizine and dimenhydrinate versus betahistine dimesylate in the treatment of Meniere's disease: a randomized, double-blind, parallel group clinical study. AB - In a randomized, double-blind clinical study, we evaluated the efficacy and tolerability of the fixed combination of cinnarizine, 20 mg, and dimenhydrinate, 40 mg (Arlevert [ARL]) in comparison to betahistine dimesylate (12 mg) in 82 patients suffering from Meniere's disease for at least 3 months and showing the characteristic triad of symptoms (paroxysmal vertigo attacks, cochlear hearing loss, and tinnitus). The treatment (one tablet three times daily) extended to 12 weeks, with control visits at 1, 3, 6, and 12 weeks after drug intake. The study demonstrated for both the fixed-combination ARL and for betahistine a highly efficient reduction of vertigo symptoms in the course of the 12 weeks of treatment; however, no statistically significant difference between the two treatment groups could be established. Similar results were found for tinnitus (approximately 60% reduction) and for the associated vegetative symptoms (almost complete disappearance). Vestibulospinal reactions, recorded by means of craniocorpography, also improved distinctly, with a statistically significant superiority of ARL versus betahistine (p < .042) for the parameter of lateral sway (Unterberger's test). The caloric tests (electronystagmography) showed only minor changes for both treatment groups in the course of the study. A statistically significant improvement of hearing function of the affected ear (p = .042) was found for the combination preparation after 12 weeks of treatment. The tolerability was judged by the vast majority of patients (97.5%) in both groups to be very good. Only one patient (betahistine group) reported a nonserious adverse event, and two betahistine patients did not complete the study. In conclusion, the combination preparation proved to be a highly efficient and safe treatment option for Meniere's disease and may be used both in the management of acute episodes and in long-term treatment. Efficacy and safety were found to be similar to the widely used standard therapy with betahistine. PMID- 14763224 TI - Doppler ultrasonography in tinnitus patients. AB - Tinnitus can be functionally measured and localized through sensorimotor and neurosensory tests. According to our Neurofisiologia Otooftalmologica data bank, 60% of patients requiring a consultation have a history of cardiocirculatory disorders. This figure has moved us to study cerebrovascular processes in those patients seeking consultation for a tinnitus symptom only. The sudden appearance of the tinnitus symptom alone independent of patient age, leads us to consider a pathology of vascular origin, which should be evaluated and treated immediately. A mistake or delay in the diagnosis could cause the symptom to become chronic, owing to the appearance of microlesions at any level of the auditory pathway, as has been demonstrated in former investigations. PMID- 14763225 TI - Vertigo as a prognostic sign in sudden sensorineural hearing loss. AB - Several prognostic indicators of favorable outcome in idiopathic sudden sensorineural hearing loss (ISSHL) have been proposed: an initial profound hearing loss, a down-sloping audiometric curve, advanced age, and the presence of vertigo. The latter has been disputed in the literature. The present study addressed the correlation between vertigo and outcome of ISSHL. Sixty-seven patients with ISSHL, aged 52 years on average, were treated with tapered doses of oral prednisone and complete bed rest. Factors found to have a statistically significant prognostic value as indicators of outcome in ISSHL were tinnitus on admission (positive indicator, p < .041) and a descending type of audiometric curve (negative indicator, p < .009). The presence of vertigo was also found to be significantly correlated with the lack of improvement in hearing, but only at the 8-kHz frequency. Dizziness was reported by 27% of the patients. The correlation between the presence of vertigo and persistent high-frequency ISSHL may be explained by the anatomic contiguity of the basal turn of cochlea and the vestibule. PMID- 14763226 TI - Temporomandibular joint dysfunction in whiplash injuries: association with tinnitus and vertigo. AB - Temporomandibular joint dysfunction in whiplash injuries is usual. The author describes the mechanism of this joint dysfunction and the physiopathology of tinnitus and associated balance disorders. PMID- 14763227 TI - Importance of the Neurootological and Equilibriometric Society. PMID- 14763228 TI - Response of human skull to bone-conducted sound in the audiometric-ultrasonic range. AB - Some new therapies for tinnitus employ bone-conducted sound in the high audio and ultrasonic frequencies, but there has been little previous research on sound transmission through the head at these frequencies. The vibrational characteristics of two dry skulls, in comparison to a live human head, were measured in the 2- to 52-kHz range. White noise was played and received through piezoelectric transducers and was Fourier-analyzed. Complex resonances and antiresonances were found in both the dry skulls and the live head and varied with small changes in the position of the transducers. There were also pronounced differences between the skulls. In comparison to the skulls, the live head showed greater attenuation and less prominent resonances and antiresonances, reflecting greater damping. The attenuation of the skulls and the head did not increase consistently with frequency but was dominated by resonances at a variety of frequencies. For designing high audio and ultrasonic tinnitus maskers and hearing aids, these results suggest that wide bandwidth must be used to compensate for the unpredictability of the resonances. PMID- 14763229 TI - Measurement of bone conduction levels for high frequencies. AB - For assessment of safety, it is necessary to measure the maximum possible force exerted by a bone conduction device coupled to the human head. Calibration of bone conduction hearing aids and vibrators in the audiometric range is based on measurement of acceleration and force using an artificial mastoid. Extending the measurement to the high audio range was accomplished using a live head. To assess safety of the UltraQuiet tinnitus treatment system, as an example, acceleration was measured from 5 to 20 kHz on a live human head as compared with calibrated levels at 6 kHz on an artificial mastoid and the live head. Using head acceleration and anchoring it to established calibration levels is a means of establishing clinical safety. Stimulation in the high audio frequencies at low levels was found to be safe. In contrast, stimulation with ultrasound requires more energy (approximately 75-90 dB re 6 kHz), which may increase the risk of damage to the car. PMID- 14763230 TI - Effects of discotheque music on audiometric results and central acoustic evoked neuromagnetic responses. AB - Audiograms and auditory evoked magnetic fields (AEFs) were observed in young male and female adults at different ages before and after being exposed to discotheque music for 4 hours. Sound pressure levels (SPLs) ranged from 95 dB (SPL) up to 130 dB (SPL). After exposure, subjects had temporary threshold shifts up to 20-25 dB, which almost disappeared after 2 hours. The majority of the subjects suffered from tinnitus that lasted approximately as long as the temporary threshold shift. Correspondingly, a transient delay and prolongation of the main component of the acoustically evoked magnetic field (AEF) negative wave, occurring 100 msec after stimulus (N100 m), was seen after this exposure; other components of the AEF (positive wave, occurring 50, 160, and 200 msec after stimulus [P50 m, P160 m, and P200 m, respectively]) occurred less often as compared to nonexposed controls. Because effects of vigilance on the AEF could be excluded, these changes can be related to the loud music, indicating an influence of noise on central auditory processing. The transient tinnitus could be caused by acoustic microinjuries (hidden acoustic predamage) of outer hair cells, leading to the persistent hearing threshold shifts from which many young adults aged 20-24 years are suffering. Occurrence of tinnitus closely coincides with the changes in hearing threshold and AEF, thus, a limitation of loudness in discotheques is needed to prevent this kind of hearing hazard. PMID- 14763231 TI - Tinnitus in childhood. AB - All of 1,420 children seen for clarification of a hearing disorder or to follow up for known difficulty in hearing were questioned as to whether they experienced tinnitus. The interview was carried out after a hearing test was conducted, which was based on play audiometry or normal pure-tone threshold audiometry, depending on the age of the child. When being interviewed, 102 children reported that tinnitus had appeared or was still present. Seventy-five children (73.5%) demonstrated difficulty in hearing in one or both ears, whereas 27 children (26.5%) had normal hearing in both ears. The most frequently obtained information (29.4%) was the progression of an existing hearing loss. Meningitis is an important cause of hearing loss and of tinnitus and could be identified in 20% of our patients. We also considered as a cause of tinnitus skull or brain trauma, acute hearing loss, and stapes surgery. However, the mechanisms of tinnitus development were not immediately clear in a large proportion of the children studied: Problems included central sensory perception (14.7%) and emotional factors (11.8%). No additional information that might lead to an understanding of the hearing loss was available for 14.7% of the patients studied. Tinnitus is a frequent symptom in childhood and, because children seldom complain about their tinnitus, such hearing problems that they report must always be taken seriously. The diagnosis should exclude metabolic disturbances, possible damage to the sensory level of the central nervous system, and circulatory disturbances. In addition, the physician should always consider emotional problems and disturbances of perception. PMID- 14763232 TI - The early kinetics of gentamicin uptake into the inner ear. AB - Transtympanic gentamicin administration has become a popular modality in the treatment of Meniere's disease. This modality and other inner-ear medical therapy are gaining increased clinical and scientific attention. We previously described the kinetics and effects of gentamicin uptake into the inner ear after delivery of the medicine into the middle ear using a variety of different techniques and sustained-release modalities [1]. In our previous work, we reported an early peak perilymph concentration and the presence of intracellular gentamicin at the 4 hour time point. We also demonstrated the activation of inner-ear damage pathways at this early time point. In this report, we examine the kinetics of gentamicin at very early time points, 1 and 2 hours after administration. Healthy adult chinchillas underwent implantation of middle-ear sustained-release devices (one to each ear) containing gentamicin. The animals then were maintained in a neutral position and underwent perilymph gentamicin sampling at the two predetermined time points. This technique allowed us to assess accurately very early time point inner-ear gentamicin kinetics and to compare the activity. The samples then were run for concentration using mass spectrometry. The information gained from this study may increase our scientific understanding about the effects of gentamicin on the inner ear and may allow clinicians to treat patients more effectively for inner-ear disorders. PMID- 14763234 TI - Characteristics of tinnitus and etiology of associated hearing loss: a study of 123 patients. AB - The aim of this study was to highlight the clinical characteristics of tinnitus and to attempt a quantitative assessment in relation to any underlying etiologies. We undertook to study a population of 123 patients attending a tinnitus clinic between 1998 and 2000. Their answers on a questionnaire allowed detailed evaluation of the characteristics of tinnitus, including such variables as the circumstances in which the tinnitus was first noticed and evaluation of its intensity and frequency. The patients each underwent a full neurootological examination with the aim of diagnosing an etiology. The great majority of tinnitus patients had an endocochlear deafness and, among these patients, acoustic trauma, endolymphatic hydrops, and presbyacusis were the commonest diagnoses (32%, 32%, and 23%, respectively). Of these patients, 93.7% with noise trauma an 86.9% with presbyacusis described their tinnitus as a stable, high pitched whistling. Those patients with active Meniere's disease or Meniere's-like syndrome described a low-pitched buzzing tinnitus. Analysis of those patients with a stable high-pitched tinnitus associated with a high-frequency hearing loss shows a statistically significant correlation between the elevation of the audiometric thresholds and the loudness of the tinnitus. For a large majority of patients with tinnitus, therefore, audiometry provides an indirect test for evaluating the tinnitus. PMID- 14763233 TI - GABAA-benzodiazepine-chloride receptor-targeted therapy for tinnitus control: preliminary report. AB - Our goal was to attempt to establish neuropharmacological tinnitus control (i.e., relief) with medication directed to restoration of a deficiency in the gamma aminobutyric acid-benzodiazepine-chloride receptor in tinnitus patients with a diagnosis of a predominantly central type tinnitus. Thirty tinnitus patients completed a medical audiological tinnitus patient protocol and brain magnetic resonance imaging and single-photon emission computed tomography of brain. Treatment with GABAergic and benzodiazepine medication continued for 4-6 weeks. A maintenance dose was continued when tinnitus control was positive. Intake and outcome questionnaires were completed. Of 30 patients, 21 completed the trial (70%). Tinnitus control lasting from 4-6 weeks to 3 years was reported by 19 of the 21 (90%). The trial was not completed by 9 of the 30 (30%). No patient experienced an increase in tinnitus intensity or annoyance. Sequential brain single-photon emission computed tomography in 10 patients revealed objective evidence of increased brain perfusion. Patients with a predominantly central type tinnitus experience significant tinnitus control with medication directed to the gamma-aminobutyric acid-benzodiazepine-chloride receptor. PMID- 14763235 TI - Psychiatric disturbances and psychotropic drugs in tinnitus patients. AB - At the ear, nose, and throat clinic of the Third Medical Faculty at Charles University and at the Psychiatry Department of the Institute for Further Education of Physicians in Prague, we examined 25 patients with tinnitus accompanied by psychiatric disturbances and followed them up for at least 6 months. Psychological changes in tinnitus patients occur in a characteristic chronological succession and manifest in three stages: defense, the search for adaptive mechanism, and relative recovery or a development of psychiatric disturbance (or both). Some psychotropic drugs probably diminish tinnitus by the mechanism of improving general psychiatric comfort and direct central influence. Subjective heaviness of tinnitus is defined by the authors as all the difficulties that exist along with tinnitus. It encompasses subjective evaluation of tinnitus loudness, frequency of tinnitus episodes, and psychiatric disturbances arising from the individual discomfort caused by tinnitus. The authors selected the visual analog scale as an easily used instrument appropriate for the complex evaluation of subjective heaviness of tinnitus. PMID- 14763236 TI - Clonazepam in the pharmacological treatment of vertigo and tinnitus. AB - We carried out a retrospective survey of 25 years of clinical experience with the use of clonazepam as a vestibular and tinnitus suppressant in the pharmacological treatment of vestibular or cochleovestibular disorders due to different causes. We reviewed the medical records of 3,357 outpatients treated with a 0.5 or 1.0-mg daily dosage of oral clonazepam during 60-180 days. Complete or substantial control of vertigo or nonvertiginous dizziness was achieved in 77.4% of the vertigo patients. Tinnitus was improved in 32.0% of the tinnitus patients. Light or mild drowsiness, depression, nightmares, or lowering of libido, reported by 16.9% of the patients as adverse side effects, tended to subside with continued therapy. We concluded that clonazepam is a very useful and safe drug for the symptomatic treatment of patients suffering from cochleovestibular disorders. PMID- 14763238 TI - Slow auditory evoked potentials: the end of malingering in audiology. AB - The application of slow vertex response audiometry (cortical evoked response audiometry), mainly in the diagnosis of pseudohypoacusis, is reported. This procedure is of interest in forensic audiology. PMID- 14763237 TI - A new therapeutic procedure for treatment of objective venous pulsatile tinnitus. AB - Pulsatile tinnitus usually means a turbulent flow within the vessels. We describe a 54-year-old man with a disabling objective pulsatile tinnitus due to a diverticulum of the sigmoid sinus toward the ipsilateral mastoid. We performed a surgical intervention via the endovascular route using coils to obliterate the diverticulum and a stent to avoid coil migration. The patient had a complete and immediate remission of the pulsatile tinnitus. This procedure has been described only in arterial circulation. PMID- 14763239 TI - Interaction between rhinitis and asthma: state of the art. AB - Rhinitis and asthma are very prevalent allergic disorders with comorbid features, similar risk factors, and environmental triggers. Pathophysiological processes are linked via tissue histopathology, immunologic pathway, and inflammatory mediators. Allergen challenge of the upper airway can increase lower-airway responsiveness and allergen challenge of the lower airway can lead to upper airway inflammation. Both allergic rhinitis and asthma exert a high social and economic burden in significant loss of work and school days as well as impairment for children and adults. PMID- 14763240 TI - The asthma and allergic rhinitis link. AB - During the past 10 years, our understanding of asthma and allergic rhinitis (AR) has evolved. The historic perspective of these allergen-induced disorders as distinct and separate entities is being displaced by current thinking that they are described better as a continuum of inflammation involving one common airway. Therefore, traditional therapies originally indicated for AR and asthma are being reassessed to explore their potential value in both upper- and lower-airway diseases. Recently, there has been a renewed interest in the role that histamines play in lower-airway disease, and interest is increasing in the leukotrienes (LTs), which are far more potent inflammatory mediators than histamines, and the role they play in upper-airway disease. Given the pivotal role that LTs play as potent inflammatory mediators in the pathophysiological state of inflammation of both airways, LT receptor antagonists recently have emerged as important therapeutic advances that have potential clinical value in both asthma and allergic rhinitis. The prevalence of asthma and AR is increasing in the general population, and a high proportion of new patients have coexisting upper- and lower-airway disease. Estimates show that 60-78% of patients who have asthma have coexisting AR. 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 may maximize benefits and outcomes for patients with concomitant asthma and AR. PMID- 14763241 TI - Therapy in the management of the rhinitis/asthma complex. AB - Allergic rhinitis is a risk factor for the development of asthma, and, conversely, asthma often is present in patients with rhinitis (17-25% in children and 20-50% in adults). Up to 80% of patients with asthma have allergic, nonallergic, or mixed rhinitis. Gastroesophageal reflux can be identified in 25 50% of patients with asthma and may be asymptomatic. Topical nasal corticosteroids typically reduce rhinitis symptoms more effectively than oral or topically administered histamine 1 antagonists but are similar in terms of ocular symptom reduction. The leukotriene D4 antagonist montelukast, as well as loratadine (29%), has been found to reduce nasal symptoms (27%) but the combination (33%) provided little additional benefit. Subcutaneous injections with a monoclonal anti-immunoglobulin E antibody for ragweed or birch allergic rhinitis have produced few anaphylactic reactions but when reactions occur, they appear 90-120 minutes after the injection. In the patients who received 300 mg of omalizumab every 3 or 4 weeks for ragweed allergic rhinitis, there were 23% fewer mean nasal symptoms than in placebo-treated subjects. In that study, antihistamines but not nasal corticosteroids were used during the study period. Overall, 70.7% of patients reported treatment as good or excellent compared with 40.8% in placebo-treated patients. The impact of omalizumab or other anti immunoglobulin E therapies on rhinitis and asthma is being investigated. In patients experiencing acute, purulent, rhinosinusitis, treatment with a nasal corticosteroid helps relieve symptoms sooner than antibiotic and decongestant therapy alone. Treatment of rhinitis or rhinosinusitis and gastroesophageal reflux should be part of the management of patients with asthma. PMID- 14763242 TI - Immune deficiency: office evaluation and treatment. AB - Congenital deficiencies of the immune system occur in children or adults and can cause severe or recurrent infections. The overall incidence of these immunodeficiency diseases is estimated at approximately 1 in 10,000, excluding selective immunoglobulin A deficiency, but this estimation is based on population studies, not hospital or clinic populations. The majority of immune defects involve antibody production; these immune deficiencies are found more often in adults than infants and children. In an allergy practice, recurrent infections are common, and determining if an immune defect is likely to be present can be problematic. Some guidelines concerning the clinical presentation and laboratory evaluation and treatment options can aid the practicing clinician. PMID- 14763243 TI - The idiopathic hypereosinophilic syndrome. AB - Idiopathic hypereosinophilic syndrome (IHES) is characterized by sustained (> 6 months) peripheral eosinophilia (> 1500 cells/mm3), organ damage, and exclusion of other causes. IHES is clinically and pathogenetically heterogenous, and several potential disease mechanisms have been described. The prognosis for patients with IHES has improved with targeted therapies and aggressive management of complications. We report a case of IHES and review the clinical features, evaluation, and options for treatment. PMID- 14763244 TI - Identification of masqueraders of autoimmune disease in the office. AB - There are several rheumatologic and autoimmune disorders that can masquerade as allergic disease. Identification of these conditions in an office setting can be a challenge for the practicing allergist-immunologist. These conditions include rheumatoid and juvenile arthritis, Sjogren's syndrome, systemic lupus erythematosus, Behcet's and antiphospholipid syndromes, systemic sclerosis, vasculitis, sarcoidosis, chronic fatigue syndrome, and fibromyalgia. The article will address these topics and include clinical uses of immunologic tests for diagnosis. PMID- 14763245 TI - Specific inflammatory cell types and disease severity as predictors of postsurgical outcomes in patients with chronic sinusitis. AB - Chronic sinusitis is a common condition that is frequently refractory to medical and surgical intervention. It has not been well defined as to which patient subgroups benefit from surgical intervention and which do not. Our purpose is to determine whether the presence of certain inflammatory cell types is predictive of postoperative outcome in sinus surgery for chronic sinusitis. We performed a retrospective chart review of 75 patients who underwent endoscopic sinus surgery or polypectomy between 1994 and 1996. Of these patients, 15 met inclusion criteria for chronic sinusitis, asthma requiring inhaled steroids, and 1 year preoperative and postoperative management by the allergist or otolaryngologist at Northwestern Medical Faculty Foundation. Ten of these 15 patients had a complete set of data allowing immunohistochemical analysis. Sections of sinus tissue obtained at surgery were hematoxylin and eosin stained and evaluated for the presence of lymphocytes, plasma cells, eosinophils, and macrophages. Immunostains for T lymphocytes, B lymphocytes, and macrophages were performed also. The total number of antibiotic courses patients received were enumerated into 6-month blocks for 1 year preoperatively and 1 year postoperatively as markers of disease activity. Postoperatively, four patients improved, three patients worsened, and three patients remained unchanged, as defined by the number of antibiotic courses required 1 year postoperatively compared with 1 year preoperatively. We found no difference in the magnitude or specific type of inflammatory cells present at the time of surgery between the groups. However, when the difference between the preoperative versus postoperative antibiotic courses was compared between patients who improved versus patients who did not improve, the improved group required markedly fewer courses relative to the nonimproved group (p < 0.009). Neither total magnitude of inflammation nor specific inflammatory cell types correlated with surgical outcome in this group. The patients who did not improve postoperatively had a statistically significantly lower number of preoperative antibiotics than the patients who improved. These findings suggest that patients with less severe disease may be less likely to benefit from sinus surgery. PMID- 14763247 TI - Strategies to reduce hospitalizations for asthma in adults. PMID- 14763246 TI - Risk factors associated with hospitalizations for asthma attacks in Turkey. AB - The detection of factors associated with hospitalizations for asthma attacks should have a great value in the development of intervention strategies. However, these factors are unknown in Turkey. Our aim was to investigate the factors associated with hospital admissions by comparing hospitalized patients with the community control asthma patients and the relationships between serum eosinophilic cationic protein (ECP) levels and the disease severity. Eighty-one subjects hospitalized with asthma (69 women and 12 men) and 300 community control asthma patients (227 women and 73 men) were enrolled in this cross-sectional study. A questionnaire including detailed demographic and clinical data was compiled by all patients. Serum ECP levels were measured in 76, 14, and 9 patients of community control, hospitalized asthma patients, and healthy controls, respectively. Hospitalized patients were older and had longer asthma duration (p < 0.001). The significant risk factors for hospital admission for acute asthma attacks were previous severe asthma (odds ratio [OR], 12.26; 95% confidence interval [CI], 5.17-29.0), aspirin (acetylsalicylic acid) and nonsteroidal anti-inflammatory drug intolerance (OR, 3.63; 95% CI, 1.70-7.74), chronic rhinosinusitis (OR, 2.24; 95% CI, 1.16-4.33), lower educational level (OR, 2.24; 95% CI, 1.33-4.18), and lower atopy ratio (OR, 1.99; 95% CI, 1.13 3.50). These parameters were similar in patients who were hospitalized and in patients who had severe asthma of the community control. ECP levels were significantly higher in hospitalized and severe asthma patients compared with healthy controls. In conclusion, the factors associated with hospitalizations were advanced age, prolonged asthma duration, presence of severe asthma, "nonatopy," acetylsalicylic acid-nonsteroidal anti-inflammatory drug intolerance, sinusitis, and lower educational level. Further intervention strategies are needed to address these markers to prevent hospitalizations from asthma attacks. PMID- 14763248 TI - Asthma hospitalizations in children: what is the solution? PMID- 14763249 TI - [Spastic syndrome and main directions of its treatment]. PMID- 14763250 TI - [Gender dimorphism in Parkinson's disease]. AB - The article addresses the gender differences in Parkinson's disease. Neurohumoral mechanisms of protective action of estrogen are discussed. An analysis of 159 patients with parkinsonism revealed definite differences in symptoms presentation and autonomic appearances in men and women that suggested a role of gender dimorphism in the development and course o Parkinson's disease. PMID- 14763251 TI - [Mental disturbances in heroin addicts in pre-recurrence period]. AB - Using a questionnaire of J. Cacciola et al (1977), 80 heroin addicts aged 16-45 years (65 males, 15 female) admitted to hospital in intoxication phase or in withdrawal syndrome, were examined. A statistical analysis of the results confirmed the existence of some mental disorders, the development of which indicated high probability of breaking the abstinence by a patient and disease relapse. These are depressive syndrome appearances, anxiety disorders, dysphoria and asthenia disorders. Using Shennon criterion, an extent of informativeness (significance) for some symptoms, relating to recurrence of driving for heroin, was determined. In most cases, symptom-targeted therapy reduced the driving thus preventing a disease relapse. PMID- 14763252 TI - [Efficacy of combined kineso- and psychotherapy in the treatment of patients with migraine]. AB - The study aimed at determining the efficacy of combined kineso- and psychotherapy in the treatment of patients with migraine in between-attacks periods. Forty-one patients with migraine (3 males, 38 females, mean age 38.2 +/- 1.5 years) were examined. The patients were randomly divided into 2 age-matched groups. No significant between-group differences in the intensity and duration of migraine attacks, autonomic and psychological status and level of tolerability to physical loading were found. The patients of group 1 (23) recurred individual kineso- and psychotherapy, and patients of group 2 (18) received a course of standard physical therapy and explanatory psychotherapy. After the treatment course, a significant decrease in intensity of headaches of tension, increased bicycle exercise tolerance, equalizing of the ratio between activities of ergo- and trophotropic systems, sympathic and parasympathic tones evaluated by mathematical analysis of heart rhythm variability, a decrease of the level of emotional tension measured by psycho diagnostic testing were observed in both groups. In the patients of group 1, global estimation of therapeutic effect (2.7 +/- 0.2) was significantly higher (p < 0.003), comparing to those of group 2 (1.8 +/- 0.2). Two months after treatment finishing, there was a reduction of migraine attacks frequency in both groups without significant between-group differences in regression extent of this index. The results obtained indicate efficacy of combined physical therapy and psychocorrection in the treatment of migraine patients. Moreover, individual approach to kineso- and psychotherapy improves the immediate results of the treatment. PMID- 14763253 TI - [The influence of the hormones of hypothalamo-pituitary-adrenal, renin angiotensin, and thyroid hormonal systems on formation of vascular encephalopathy]. AB - The aim of the study was to investigate the concentration changes of free (f) thyroxine (fT4), triiodothyronine (fT3), thyrotrophic hormone (TTH), adrenocorticotrophic hormone (ACTH), aldosterone and renin in the blood plasma of patients with vascular encephalopathy (VE) and to evaluate their influence on formation of chronic blood circulation deficiency. Thirty-nine patients with VE of stages I and II, aged 45-73 years, without current acute or chronic (in exacerbation stage) other somatic diseases have been studied. The results revealed that a different extent of diffuse lesion of brain tissue was accompanied by an own pattern of thyroid homeostasis: 1) significantly higher TTH levels with intact regulatory negative feedback "fT3-TTH" in patients with VE, stage 1; 2) significant TTH concentration decrease, with pronounced suppression of thyroxine conversion to triiodothyronine and interaction disturbance in the "fT3--pituitary gland" system, in patients with VE in stage II. A distinct trend towards plasma cortisol level increase as well as significantly lower active plasma renin levels were found in the patients with VE in stage II comparing to those in stage I. Correlation analysis demonstrated the absence of an interaction between patient's age and hormonal homeostasis state and an extent of vascular stenosis. The data suggest involvement of the hormones of hypothalamopituitary adrenal, thyroid and renin-angiotensin systems in VE formation thus enabling using of the results of their testing as an additional criterion for estimation of brain diffusive lesion severity. PMID- 14763254 TI - [Neuroimmunopathological mechanisms of preclinical epileptogenesis and their correction by cycloferon]. AB - The peculiarities of immunologic processes and immunocorrecting potential of Cyclopheron were studied in 62 patients with preclinical epileptogenesis. Using clinical, EEG and biochemical examination, expression of epileptogenesis was estimated as regressed, compensated, increasing, decompensated. Preclinical stage of epilepsy was accompanied by disturbance of immunological brain reactivity the expression of which was correlated with a level of compensative epileptogenesis manifestation. The decrease of the latter is followed by intensive disintegration of neuroimmune interactions characterized by immunodeficiency of mixed type with significant presentation of neuroimmune disturbances. Therefore, a use of Cyclopheron, as a medication with multilevel immuno-correcting action, is expedient at initial stage of epilepsy. PMID- 14763255 TI - [Amphetamine action on morphochemical brain organization]. AB - The activity of enzymatic systems--monoamine oxidase, types A and B, and acetylcholinesterase involved in neuromediator utilisation and their levels correlation in the cortex and caudate nucleus brain tissue subfractions in control and during long (3 weeks) amphetamine (psychostimulator) injections in dosage 2.5 mg/kg were studied. Wistar and August rats different by behavioral characteristics were used. Differences of enzyme activity on subcellular level both in controls and in rats with dopaminergic dysfunction, caused by amphetamine, were found. Comparing to Wistar rats, in August ones characterized by lower motor activity and elevated stress sensitivity, amphetamine caused a significant MAO A and acetylcholinesterase activity inhibition, the changes being more pronounced in the caudate nucleus. It is suggested that the differences of enzymatic activity, reflecting the state of brain neuromediator metabolism and emerging in pathological conditions, may underlie a mechanism of CNS psasticity. PMID- 14763256 TI - [The tendencies of populations' psychological health in a large agri-industrial region]. PMID- 14763257 TI - [Current approaches to the treatment of vertigo in children]. PMID- 14763258 TI - [Personality features of patients with systemic rheumatic disorders]. PMID- 14763259 TI - [Efficacy of depakine chrono and lithium combination in the treatment of manic states]. PMID- 14763260 TI - [Topamax in the treatment of symptomatic frontal epilepsy]. PMID- 14763261 TI - [Hysteria and multiple sclerosis]. PMID- 14763262 TI - [Galantamine (reminyl) in the treatment of Alzheimer's disease and vascular dementia]. PMID- 14763263 TI - [Diabetes mellitus and depression]. PMID- 14763264 TI - [Is psychosurgery expected back?]. PMID- 14763265 TI - [On the article by I. I. Mikhailova, V. S. Iastrebov and S. N. Enikolopov "Clinico-psychological and social factors influencing stigmatization of the mentally ill patients from different nosologic groups"]. PMID- 14763267 TI - [The 1st Russian International Congress: Cerebrovascular Pathology and Stroke]. PMID- 14763266 TI - [All-Russian conference "Modern problems of biological psychiatry and narcology"]. PMID- 14763268 TI - HMOs would be wise to investigate alternative ways to improve health. PMID- 14763269 TI - Do the Medicare math, then make quick decision. PMID- 14763270 TI - Compensation monitor. Opportunities abound for physician executives. PMID- 14763271 TI - Health plan 2009. The enduring problem of rising costs. PMID- 14763272 TI - Health plan 2009. Consumer-directed health care won't fly. PMID- 14763273 TI - Health plan 2009. Technology's good and ill effects. PMID- 14763274 TI - Health plan 2009. No end in sight to medical inflation. PMID- 14763275 TI - Health plan 2009. Rise of nonphysician clinicians. PMID- 14763276 TI - Health plan 2009. Health care needs to catch up. PMID- 14763277 TI - Health plan 2009. 'A final underwriting death spiral'. PMID- 14763278 TI - Cardiologists call collaboration heart of effort to improve care. PMID- 14763279 TI - Transparency called key to uniting cost control, quality improvement. PMID- 14763280 TI - Reinsurers offer services to keep client costs down. PMID- 14763281 TI - The formulary files. Increasing drug copayments deters compliance. PMID- 14763282 TI - Logic flies when EMRs debated. PMID- 14763283 TI - Choking off a tumor's blood supply. PMID- 14763284 TI - Major legislative reform not in store for 2004. PMID- 14763285 TI - The path of enlightenment. Independent contracting may be the answer. PMID- 14763286 TI - Building a better world. PPW's sojourn to South America. PMID- 14763287 TI - From the nurse's mouth. One clinician's view on reprocessing. PMID- 14763288 TI - [Maternal and infant prognosis of emergency cesarean section: prospective study of the Principal Hospital in Dakar, Senegal]. AB - The prognosis of emergency cesarean section is poor for both the mother and child in developing countries. The respective impact of obstetrical and surgical factors has rarely been analyzed. This prospective study was carried out in 370 women (mean age, 30.5 years) who underwent emergency cesarean section at Principal Hospital in Dakar, Senegal, between January 1 and December 31, 1997. Fifty percent of these women had been transferred from an outside maternity clinic. Indications related to the mother (75% of cases) or fetus (25% of cases) were divided into two groups according to degree of emergency: absolute (n = 163) and relative (n = 207). Placental hematoma (n = 64) and fetus-pelvis size mismatching (n = 49) were the main indications in both groups. The technique chosen for initial anesthesia performed by a specialized nurse in most cases was either spinal anesthesia if there were no contraindications (50.8%) or general anesthesia (49.2%). There were 5 complications including 1 that was fatal (aspiration during intubation for general anesthesia). The postoperative maternal morbidity rate was low (n = 7) and outcome was favorable. A total of 7 patients (1.9%) died due to anesthesia-related events in 1 case and obstetrical factors in 6. Mortality in the absolute emergency group was significantly higher for women who were transferred from other clinics (p < 0.02). Child mortality (n = 87) occurred prior to delivery in two thirds of cases and after delivery in one third. Child mortality was significantly higher in the absolute emergency group (RR = 5.4; IC95% = 3.2-8.9, p < 10(-6)). Mother and child mortality rates were correlated with the severity of obstetrical manifestations and delay of care. Findings also showed that a well-organized care system lowers the operative risk of emergency cesarean section even in developing countries. PMID- 14763289 TI - [Maternal mortality in developing countries: statistical data and improvement in obstetrical care]. AB - Since launching of the safe motherhood initiative in 1987, much work has been undertaken, understanding of the situation in developing countries has improved, and numerous health programs have been designed. However the end result of action has been considered disappointing more often than encouraging especially in Sub Saharan Africa. What is the true picture? The purpose of this article is to review the means available for studying all facets of maternal mortality and methodological precautions that must be applied in the interpretation of statistical data. Perusal of recent reports on maternal mortality reveals that estimated incidences in different populations vary widely from 85 to 1000 per 100,000 live births, that rural zones are more affected than urban areas, that reductions have been achieved in the major cities, that the most common direct obstetrical causes are postpartum hemorrhage, dystocia with uterine rupture, eclampsia, and sepsis, and that 70% of deaths are avoidable, i.e., due to absent or insufficient care. Although currently underused, qualitative study methods are gradually being implemented and will identify the health care sectors requiring priority improvement. Based on previous experience, it is unlikely that technical or obstetrical measures and action on the part of medical professionals alone will achieve any reduction in maternal mortality without the commitment of political authorities. PMID- 14763290 TI - Perinatal mortality and morbidity in developing countries. A global view. AB - Global perinatal mortality figures show that of the 132 million births per year, there are between 6 and 7 million perinatal deaths. While 90% of these births are in less developed countries, perinatal deaths take 98% of the global share. These statistics show on average the rates as they were in England during the 1930s. The most common recorded medical causes of perinatal deaths are also similar in the less developed countries, and the common denominators are early childbearing, poor maternal health and above all, the lack of appropriate and quality services. Although life-saving practices for most infants have been known for decades, currently a third of mothers still have no access to services during pregnancy, and almost half do not have access to services for childbirth. There are enormous variations both among and within countries. It takes innovation to find the best fit between the needs of women and infants and resources. A health worker with excellent knowledge and skills is the key resource and the best investment. The cost is moderate, and the investment pays a high dividend in improved health of both the mother and her baby, and better health for the next generation at lower cost. PMID- 14763291 TI - [Malaria during pregnancy: consequences and interventional perspectives]. AB - The impact of malaria during pregnancy varies greatly according to the intensity of transmission. Severe acute complications including cerebral malaria or materno fetal death seem to be confined to areas of unstable transmission where malaria is uncommon except during epidemics. In areas of stable endemicity, the main consequences are maternal anemia and intra-uterine growth retardation resulting in low birthweight (LBW) particularly after first pregnancies. Recent studies have demonstrated that frequency and severity of placental malaria are greater in pregnant women with concurrent HIV infection. Since 1964 several controlled trials have been conducted to evaluate chemoprophylaxis in pregnant women mainly in tropical Africa where malaria transmission is stable. Findings have usually demonstrated an increase in mean birthweight after prophylaxis especially among primigravidae. Prophylaxis also had beneficial effects on anemia. Another finding of these trials was that prevention is less effective for women with HIV co infection and that higher doses may therefore be required in such cases. In our opinion prophylaxis should be actively promoted as a routine public health measure for pregnant women in endemic areas. Current recommendations call for the use of a sulfadoxine-pyrimethamine twice or three times during pregnancy in antenatal clinics. This combination is more effective as a result of strong resistance of parasites to chloroquine. High cost and possible adverse effects in pregnant women prohibit routine use of mefloquine in developing countries. Integration of malaria prophylaxis into antenatal care services with nutrition and immunization measures should enhance the overall efficacy of prevention in outlying clinical facilities. Recent identification of molecular receptors involved in the cytoadherence of parasitized red blood cells to the placenta may lead to the development of new therapeutic or vaccinal approaches for pregnant women. PMID- 14763292 TI - [HIV-1 transmission from mother to child and its prevention]. AB - Mother-to-child transmission of HIV-1 can occur during pregnancy, labor/delivery, or breastfeeding. Without intervention HIV infection occurs in about 35% of infants born to HIV-infected women: 10% during pregnancy, 15% during labor/delivery, and 10% during breastfeeding. As early as 1994, the PACTG 076 ANRS 024 study documented the remarkable effectiveness of AZT in reducing mother to-child HIV transmission from 26% to 8% in women who did not breastfeed. Since then, AZT or multiple antiretroviral therapies has been widely used in pregnant HIV-infected women in industrialized countries resulting in decline of the transmission rate to less than 3%. In developing countries where most perinatal transmissions occur, preventive treatment is more difficult to implement. Abbreviated treatment using AZT or nevirapine (NVP) in a single dose to the mother during delivery and to the newborn reduces perinatal transmission but to a lesser extent in comparison with standard treatment. Combination treatment using AZT + 3TC or AZT + NVP is more effective. Elective cesarean section has also been proposed but is not recommended in developing countries. To date the only alternative of proven efficacy for reducing transmission during breastfeeding is formula feeding, but this method may be hazardous if sanitary conditions are poor. Studies are currently under way to test the efficacy of antiretrovirals administered to the mother and/or infant in reducing HIV transmission during breastfeeding. PMID- 14763293 TI - [Maternal mortality in developing countries: what strategies to adopt?]. AB - Despite an international consensus on the strategies necessary to achieve a massive reduction of maternal mortality and related neonatal mortality, many countries have made no progress in these areas. The main reason for this failure is that this aspect of public health and the basic human right to bear children under acceptably safe and respectable conditions have received neither sufficient attention from governments in developing countries nor long-term technical and financial support from rich countries. Yet a sound health care system that is accessible to the poorest classes is prerequisite for durable socio-economic development. Implementation of the UN Millennium Development Goals (MDG) provides an excellent opportunity to reaffirm the need for massive support of programs undertaken in this domain by developing countries and for implementation of an effective strategy to enhance access to quality care for the poorest classes. The purpose of this article is to review the main points in a strategy to reduce maternal mortality, i.e., use of practices with documented effectiveness; access to qualified personal during pregnancy and delivery; availability of health services and underlying facilities; the role of individuals, families, and communities; and the political and legal framework. This article also stresses the fact that programs designed to enhance maternal and newborn health can significantly strengthen the health care system for the community as a whole: maternal health offers a gateway for strengthening health care services in general. PMID- 14763294 TI - [Breastfeeding in developing countries: update and current recommendations]. AB - Data from numerous surveys conducted by UNICEF using standardized methods in most developing countries over the last 10 years now enable evaluation of the status of breastfeeding in the world. The purpose of this report is to present findings concerning the duration of exclusive breastfeeding, appropriate complementary feeding, and continued breastfeeding after 6 months. Analysis of these data indicates that progress is being made but that various problems remain. In an effort to improve the situation, the WHO/UNICEF has undertaken a number of initiatives and issued several recommendations. PMID- 14763295 TI - [Progress in the world health status of children during the period 1990-2000]. AB - Multiple Indicator Cluster Surveys (MICS) conducted by UNICEF using standardized methods in 66 countries have provided data for evaluating the status of child health during the period between 1990 and 2000. This report presents findings on mortality in children less than 5 years of age, malnutrition indicators, immunization status and integrated management of childhood illness. It is concluded that children at the dawn of the 21st century are exposed to new risks related to poverty, war, violence, urbanization, and VIH/AIDS. PMID- 14763296 TI - [Development of African children: influence of the home environment]. AB - The purpose of this report was to show how special features of the African home environment affect child development. Special emphasis was placed on the traditional social structure and the roles of various players. The different stages of development are described in relation to the major social events in African families and communities. Psychomotor development in African children presents several particularities in relation to children in the industrialized world but differences tend to decrease as a result of ongoing social upheaval. Modernization or globalization has had a profound effect on family organization and child care. These changes must be taken into account by officials in charge of child-psychiatry services. Preventive initiatives should taken into account and, insofar as possible, preserve traditional familial resources. PMID- 14763297 TI - [Environment and child health: from health transition to shared risk?]. AB - Children under the age of 18 account for almost half of the world's population, with most living in developing countries. Young people are especially sensitive to acute and chronic environmental conditions and 43% of environmental diseases occur in the 12% of the world's population under age 5. The main environmental threats to the health of children in developing countries are inadequate access to clean water for drinking and hygiene, exposure to air pollution: primarily indoors and secondarily outdoors, risk of accidents and wounds, and poisoning due to toxic products. Recent data suggest that the number and diversity of environmental risk factors affecting child health is increasing as a result of increasing malnutrition, pollution, and violence and consequently that the level of health and quality of life of future generations will decrease. Due to the complexity of the interactions between environmental factors and socio-economic determinants, the epidemiological transition model is poorly suited to analyzing and predicting the concurring risks of infectious disease and chronic disease (diabetes, cancer...). This article presents a number of recommendations for training health professional, developing environmental reference centers, implementing risk assessment, coordinating decentralized activities and policy, and involving parents and children in the decisional process with emphasis on divulgating study findings and developing interfaces between the various stakeholders. PMID- 14763298 TI - [Neonatal and pediatirc intensive care in developing countries. Myth or reality? Luxury or necessity? From theory to practice]. AB - Neonatal and pediatric intensive care poses a major challenge in developing countries where the socio-economic level is low and health care resources are limited. Given the large size of the pediatric population as well as of great socio-cultural and symbolic importance of the child, there is a natural, compelling need for management of serious diseases in newborns and infants. The lack of timely disease prevention and treatment accounts in part for the frequency and severity of cases. Thus the status of intensive care units can exist in this setting is a pertinent question. The purpose of this study was to attempt to answer this question by surveying neonatal and pediatric care in developing countries based on experience in Morocco over the last 25 years. PMID- 14763299 TI - [Parasitic diarrhea in eutrophic and malnourished children]. AB - Almost all children living in endemic zones are infected by gastrointestinal parasites. However only 3 to 5% develop diarrhea directly related to parasite infection. Entamoeba hystolytica and Entamoeba dispar coexist in many areas. In the past Entamoeba dispar was called non-pathogenic ameba. The vegetating forms are microscopically identical and detection of wall differences using biochemical tests is unreliable. Thus since it is rarely possible to determine whether or not a vegetating ameba found in stools is hematophagous treatment using metronidazole is the only alternative. Failure of such treatment indicates that dysentery is probably due to a cause other than amibiasis, e.g., bacterial infection in most cases. Another protozoan commonly found in endemic areas is Giardia. Giardia can cause diarrhea and this is frequently the case in undernourished children. Giardia infection leads to severe atrophic villosity requiring appropriate specific treatment. In children cryptosporidioses may be asymptomatic or lead to diarrhea especially in cases associated with malnutrition or immunodeficiency related in particular to AIDS. Helminths are a rare cause of significant diarrhea except Anguillula in undernourished children. In children presenting severe malnutrition, anguilluliasis can lead to serious consequences and requires immediate treatment using ivermectin. To avoid severe diarrhea in children presenting immunodeficiency induced by corticotherapy or chemotherapy for cancer, prophylaxis is mandatory against anguilluliasis using ivermectin and usually against giardiasis using metronidazole. PMID- 14763300 TI - [Childhood malaria in endemic areas: epidemiology, acquired immunity and control strategies]. AB - Malaria is a major cause of childhood mortality and morbidity in endemic areas. The incidence and severity of malaria depends on various entomological, parasitological, environmental, and human factors. Clinical presentation and epidemiologic features vary according to genetic factors, personal behavior, and immune status. Populations exposed to frequent infection may develop partial labile protective malarial immunity at the price of high morbidity and mortality due to ineffectiveness of antimalarials. Malarial immunity affects not only mortality and severity of malaria but also uncomplicated malarial attacks and plasmodium infection. The number of determining factors involved in relatively limited but, like a game of chess, their possible combinations are numerous. A better understanding of malaria is necessary to plan and implement effective control strategies. That is the aim of this review. PMID- 14763301 TI - [HIV-1 infection in children in African countries]. AB - The prognosis of HIV infection is dramatic for children living in poor countries. Over 50% die within two years. The World Health Organization has estimated that 1500 children will be infected daily until large-scale national programs for prevention of mother-to-child transmission are implemented. A better understanding of the causes underlying early morbidity could lead to a substantial reduction in mortality pending use of antiretroviral drugs which have demonstrated promising results in preliminary tests on children in poor countries. PMID- 14763302 TI - [Tuberculosis in African children: epidemiologic, clinical and therapeutic aspects (corrected)]. AB - In the last 10 years since world health assembly declared re-emergence of tuberculosis to be a worldwide emergency, most countries have initiated control strategies based on the recommendations of the World Health Organization. Implementation of national control programs has not only been encouraged but also become a necessity in the face of the constantly increasing number of cases and HIV epidemic. Diagnosis of tuberculosis in children is difficult and reliable estimation of prevalence is difficult. The purpose of this study was to analyze diagnostic criteria in 1128 children in Algiers, Algeria. Short-course chemotherapy was also evaluated. A prospective study in 733 children showed that 6-month regimes administered in the framework of a national program are effective and led to few complications. Short-course treatment also promotes better patient compliance. Consideration was also given to prophylactic treatment for contact children and adverse reactions to BCG. PMID- 14763303 TI - [Purulent meningitis in children: special considerations for developing countries]. AB - Bacterial meningitis is frequent in tropical zones as a result of recurring epidemics, meningococcal meningitis and near-total absence of anti-haemophilus type b vaccination. Recently the situation has been further complicated by the unexplained appearance of meningococcal W135 epidemics in Burkina-Faso. With regard to pneumococcal meningitis, the appearance of antibiotic strains is a worldwide phenomenon that has greatly increased the cost of antimicrobial treatment. Hopefully in the future children in tropical areas will be able to benefit from immunization using a conjugate vaccine combining anti-haemophilus b, meningococcal type a and W135 vaccination. In this regard the currently available 7 valence pneumococcal vaccine is poorly suited to tropical epidemiology and that an additional 4 serotypes would be needed to achieve 85% coverage. PMID- 14763304 TI - [Update on oral rehydration salt solutions used for treatment of childhood diarrhea]. AB - After 20 years of research to improve oral rehydration salts (ORS), a new formula has been developed and is now recommended by the World Health Organization (WHO) and the United Nations International Children's Emergency Fund (UNICEF). The new formula that contains slightly less sodium and glucose than previously not only decreases the volume of diarrhea and vomiting in children presenting acute non cholera-related diarrhea but also, and above all, significantly reduces the need for intravenous fluid treatment. This new so-called reduced osmolarity formulation has been included in the WHO's list of essential drugs. PMID- 14763305 TI - [Antiparasitic treatments in pregnant women and in children in 2003]. AB - Like antibacterial agents, antiparasite drugs for pregnant women and children must be chosen in function of the stage of pregnancy, age of the child, and expected benefit-risk ratio. While no agent is totally safe, there are few absolute contraindications. Most zones of serious endemic parasite disease are located in developing countries where parasite, bacterial, or viral conditions combined with poor nutrition treatment make it necessary to treat disease in a complex pathogenic environment that weakens pregnant women and children with multiple parasite infections. In both temperate and tropical zones, there have been few real therapeutic advances involving release of new products on the market or development of new indications for existing products. Constant appearance and extension of hematozoa resistance to conventional and even more recent antimalarial agents have prompted research to find new active drugs and long-lasting treatment combinations. Real therapeutic breakthroughs have resulted from the need to develop safe drugs without substantial side-effects for single dose use in control programs against endemic parasite diseases in mass populations including pregnant women and young children in tropical zones. There are several notable examples in the field of major verminous diseases. Ivermectin is a versatile drug that can be used against filariasis as well as for management of intestinal worms or ectoparasitosis in temperate and tropical countries. Praziquantel is an important advance in platyhelminthiasis, especially bilharziais. Triclabendazole, the latest addition to the benzimidazole family, has shown promise as a substitute for bithionol, that is difficult to procure and not recommended in pregnant women, for treatment distomiasis occurring in pregnant women and children. Other examples include albendazole against giardiasis, nitazoxamide against cryptosporidiosis, artemisinine against bilharziasis, and paramomycine, not recommended in pregnant women, against leishmaniasis. PMID- 14763306 TI - [Maternal and childhood immunization in developing countries: success, problems and new approaches]. AB - A quarter century after implementation of the Expanded Program of Immunization by the WHO, partner organizations and member states, significant progress has been made towards eradication of poliomyelitis, elimination of neonatal tetanus, and control of measles. Problems remain due to difficulties involved in extending immunization coverage to some regions (Africa), integrating new vaccines into routine immunization schedules and securing sufficient funding for programs. Injection safety is also a major problem that should be resolved by utilization and proper disposal of single-use auto-disabling syringes. The forthcoming availability of new vaccines on the market and the action of the Global Alliance for Vaccines and Immunization hold reasonable hope for the future. PMID- 14763307 TI - [Evaluation of long-term outcome of Senegalese children sent abroad for cardiac surgery]. AB - The incidence of childhood heart disease in developing countries is high, but access to cardiac surgery is limited. This mismatch has given rise to numerous humanitarian programs aimed at sending children abroad for surgical treatment. However little is available about the long-term outcome of these interventions. In 1999 we conducted a retrospective study of 168 Senegalese children undergoing follow-up at the Principal Hospital in Dakar after being transferred to Europe or the Ivory Coast for surgical treatment thanks to the Terre des Hommes Association. A total of 85 children presented congenital heart disease (CHD) and 83 presented acquired heart disease (AHD). Fifteen patients did not undergo surgery due to either contraindications or preoperative death. At the end of study, 23 children had been lost to follow-up mostly from the CHA group and presumably some were cured. Outcome was verifiable in the remaining 145 patients with a median follow-up of 5.6 years. Ninety-seven patients were cured or undergoing surveillance. Quality of life was better in the CHD group (p = 0.047). Forty-eight patients died including 16 in the CHD group and 32 in the AHD group. Perioperative mortality (n = 19) was lower and late mortality (n = 29) was higher in the AHD group (p = 0.005). In the AHD group compliance with surveillance was better for children with valve prostheses. In children treated for isolated mitral valve insufficiency, late mortality was higher after valve replacement than valve repair (p = 0.04). In absence of comparative study data, high mortality was due in part to the long delay between the decision to send the patient abroad and the actual evacuation. These findings support humanitarian action to promote cardiac surgery in developing countries. PMID- 14763308 TI - [Management of children with sickle cell disease in Africa: experience in a cohort of children at the Royal Albert Hospital in Dakar]. AB - Management of major sickle hemoglobinopathies in industrialized countries has improved significantly over the last few years thanks to strict application of the preventive and curative measures developed as a result of a better understanding of the underlying pathophysiological mechanisms. However patients in Africa have benefited little from progress in the field due to the lack of human and physical resources in sharp contrast with the high prevalence of the disease. The purpose of this study was to analyze problems involved in management of childhood sickle cell disease in Africa based on our experience in a cohort of 556 cases treated over a period of 12 years. The main problems were the same as those encountered in other black African nations, i.e., delayed diagnosis due to a lack of routine neonatal diagnostic screening, difficulty implementing anti infectious prophylaxis due not only to the high cost of recommended vaccinations not covered by the Expanded Program on Immunization but also to poor compliance with antiobioprophylaxis, and insufficient transfusion facilities hindering application of long-term transfusion protocols when indicated. In addition the high prevalence of digestive-tract parasitosis and malaria raise the need to combine standard preventive measures with routine parasiticidal treatment and malarial prophylaxis adapted to each geographical area. The high frequency of associated iron deficiency requires systematic laboratory testing to identify and treat resulting manifestations during follow-up. An important prerequisite for widespread implementation of appropriate preventive and curative measures in Africa is recognition of sickle cell disease as a priority in public health care policy. PMID- 14763309 TI - [Epidemiology and prognosis of childhood cancers in Dakar, Senegal]. AB - Childhood cancer has often been considered as a problem mainly affecting industrialized countries. In reality more than half of cases occur in developing countries where management and diagnosis are major issues. This retrospective study includes 130 children (0.3% of admissions) between the ages of 0 and 15 years hospitalized for malignant disease at the Principal Hospital in Dakar, Senegal between January 1, 1990 and December 31, 2000. Mean age was 97 months and M/F sex ratio was 1.2. Mean delay for admission was 3 months. The five most frequent cancers, accounting for 75% of cases, were leukemia (n = 28), lymphoma (n = 21), nephroblastoma (n = 21), retinoblastoma (n = 16) and osteochondrosarcoma (n = 10). Treatment was completed in 18% of cases. Half of patient were lost from follow-up. The cure rate was 10% overall and 50% for patients receiving complete treatment. The highest cure rate was achieved for nephroblastoma, i.e., 58% of cases treated. Management of childhood cancer in Africa is confronted with numerous problems, namely, paucity of specialized staff, absence of expert centers, shortage of anticancer drugs, lack of financial resources, and delay in treatment. These factors associated with frequent malnutrition and recurrent infectious diseases, greatly lower cure rates in comparison with industrialized countries. PMID- 14763310 TI - [Management of severe malnutrition in children in tropical climates]. AB - Severe malnutrition is defined as a weight for height index less than-3 standard deviations (SD) from the NCHS reference or the presence of edema. Without appropriate treatment, the risk of death is high. The WHO now proposes a standardized treatment protocol that consists of first treating initial complications and then two-phased feeding with two therapeutic formulas (milk F75 and milk F100) made of dried skimmed milk, oil and sugar highly supplemented with vitamins and minerals but containing no iron. Milk F75 featuring moderate protein and energy content is used for the first few days until associated infections are under control and anorexia disappears. Milk F100 featuring higher energy and protein content that allows high weight gain is subsequently used for nutritional rehabilitation. Milk F100 is gradually replaced by solid diets for home based nutritional rehabilitation. PMID- 14763311 TI - [Epidemiology and prognosis of childhood injuries in Dakar, Senegal]. AB - The impact of accidental injury on childhood morbidity and mortality in Africa is underestimated. The frequency and severity of accidents is high. This retrospective study includes 381 children (3% of admission) between the ages of 0 and 15 years hospitalized for accidental injury at the Principal Hospital in Dakar, Senegal, between January 1, 1997 and December 31, 2000. The annual incidence of childhood injury doubled over the 4-year study period. Mean age was 48 months and the M/F sex ratio was 1.7. The causes were trauma (n = 184) due to domestic accidents (n = 109 including 91 falls) or road accidents (n = 75), poisoning (n = 129 including 38 caustic soda burns), foreign body aspiration or ingestion (n = 30), burns (n = 18), and miscellaneous accidents (n = 20). Most fatalities (9%) were due to road accidents and falls. Rapid uncontrolled urbanization and problems of displaced rural populations in adapting to city living may account for the high frequency of falls from high buildings and road accidents. As in industrialized countries road accidents, though not the most frequent, are the most lethal. Lack of emergency services accounts for severity. Vigorous information campaigns and prevention action will be needed to lower the high morbidity and mortality of childhood injury in developing countries. PMID- 14763312 TI - [Self-destructive behavior in African adolescents]. AB - The purpose of this study was to analyze self-destructive behavior in adolescents in Senegal. Based on four case reports the authors stress the central importance of family breakdown in the process leading to this behavior as opposed to the first descriptions implicating an offense against the honor of the group. The vulnerability of teenagers in the absence of structural initiation is also advanced. Systematic intervention may be recommended in such cases. PMID- 14763313 TI - The devil is in the details. More than a decade later, the ADA is still a tricky law to follow. PMID- 14763314 TI - On resilience. PMID- 14763315 TI - Development and implementation of a nurse-managed anticoagulation program. AB - Nurse-managed anticoagulation is an emerging trend in primary care with the potential to improve the quality, efficiency, and responsiveness of outpatient anticoagulation services. Research indicates that specially trained nurses, using portable international normalized ratio (INR) monitors and decision-support systems, safely and effectively perform independent warfarin-dose adjustment. This article describes the development, implementation, and preliminary evaluation of a nurse-managed anticoagulation program as well as key lessons learned in the process. Factors for success included (a) availability of portable INR monitors, (b) designation of specially trained anticoagulation nurses, (c) use of an effective decision support system, and (d) support from staff and management. PMID- 14763316 TI - Development of relationship-centered care. AB - A review of the literature (1995-2001) was conducted on the relationship-centered care model first proposed in Health professions education and relationship centered care: A report of the Pew-Fetzer task force on advancing psychosocial education. The report describes a developmental process that occurs in four dimensions of relationship-centered care: patient-practitioner, practitioner practitioner, practitioner-community, and self-awareness. Publications were read and sorted by professional affiliation of the first authors, dimension of relationship-centered care citations from the Pew-Fetzer Task Force report, and references to activities of regional relationship-centered care networks affiliated with the Fetzer Institute. The Pew-Fetzer relationship-centered care model, when included in healthcare professional education, will improve consumer and provider satisfaction. PMID- 14763317 TI - Prevention of postoperative mediastinitis: a clinical process improvement model. AB - In a prospective study of more than 4,000 consecutive patients who underwent any cardiovascular procedure requiring sternotomy incision in a 650-bed tertiary care hospital, a gradual increase in deep sternal infections from 0.8% in 1995 to 2.1% in 1999 was noted. By using a Plan-Do-Check-Act process improvement model, several interventions to decrease the infection rate were planned and implemented based on hypotheses generated from the characteristics of infected patients. These interventions included chlorhexidine preoperative shower, discontinuation of shaving, administration of antibiotics in the holding area, segregation of instruments, and implementation of an insulin protocol. Findings included a decrease in deep sternal and leg infections after implementation of these interventions. Deep sternal infection rates decreased from 2.1% to 1.5% and leg infection rates, from 1.93% to 0.47%. Results were not statistically significant but were clinically relevant. Furthermore, a total of $200,000 was achieved in cost savings in 1 year. PMID- 14763318 TI - Continuous quality improvement for the clinical decision unit. AB - Clinical decision units (CDUs) are a relatively new and growing area of medicine in which patients undergo rapid evaluation and treatment. Continuous quality improvement (CQI) is important for the establishment and functioning of CDUs. CQI in CDUs has many advantages: better CDU functioning, fulfillment of Joint Commission on Accreditation of Healthcare Organizations mandates, greater efficiency/productivity, increased job satisfaction, better performance improvement, data availability, and benchmarking. Key elements include a database with volume indicators, operational policies, clinical practice protocols (diagnosis specific/condition specific), monitors, benchmarks, and clinical pathways. Examples of these important parameters are given. The CQI process should be individualized for each CDU and hospital. PMID- 14763319 TI - The effect of computer-assisted evaluation of labor on cesarean rates. AB - Dystocia, or slow labor, is the leading cause of first-time cesarean sections. Current diagnostic guidelines for dystocia are vague, and there is no clear postoperative confirmatory evidence to assess the correctness of this diagnosis. For several decades, various professional organizations have indicated that cesarean rates could be lowered safely and have recommended levels that are far below national averages. The three major factors, of roughly equal importance, associated with cesarean for slow labor are the baby's weight, the mother's height, and the threshold at which the physician believes it is reasonable to intervene. The last is the only modifiable factor, and quality programs are a major part of changing medical behavior. By using two study designs, the effect of a mathematical method for evaluating labor progress on the rate of cesarean section was measured. In the prospective randomized clinical trial, the relative risk of cesarean in the experimental group was unchanged at 1.04. In the pretest posttest analysis, the rates fell from 19.54% to 17.04% at 6 months and 16.62% at 12 months. PMID- 14763320 TI - Hospital accreditation and patient satisfaction: testing the relationship. AB - This article describes a study that examines the relationship between two principal measures of institutional healthcare quality: accreditation scores and independently measured patient-satisfaction ratings. This study involved a retrospective review and comparison of summative and selected categorical hospital accreditation scores from the Joint Commission on Accreditation of Healthcare Organizations and independently measured patient satisfaction ratings. A total of 41 acute care, 200-plus bed, not-for-profit hospitals in New Jersey and eastern Pennsylvania were included. Correlation and multiple-regression statistical methods were employed. The results revealed no relationship between these quality indicators on a summative level and no meaningful pattern categorical relationships. This finding suggests a disassociation between these two quality indicators, thus supporting the use of a balanced scorecard approach to hospital quality management. The study also revealed certain shortcomings in these two quality indicators, relating to insufficient score variability, which should be considered by those using such data to manage quality outcomes. PMID- 14763321 TI - International Tinnitus Forum, twentieth anniversary meeting. San Diego, California, September 21, 2002. PMID- 14763322 TI - Gene-based diagnostic and treatment methods for tinnitus. AB - The etiology of tinnitus combines hereditary and environmental factors. To help develop optimal therapies for tinnitus, it is necessary to characterize the genetic contributors to the pathophysiology and to design treatments at the level of the gene. Inner ear gene therapy involves delivery of genes into the vestibular or auditory portions of the inner ear for preventive or reparative therapies at the level of the sensory epithelium or the eighth nerve neurons. BDNF and GDNF are among the neurotrophic factors shown to be overexpressed with gene therapy and to protect the inner ear against trauma. Combined treatment with Ad.GDNF and electrical stimulation provided enhanced preservation of denervated spiral ganglion neurons. The use of viral vectors for gene therapy may involve side effects, including immune response to the viral proteins. Treatment with immunosuppressive medications can reduce the negative consequences of adenovirus mediated gene therapy. PMID- 14763323 TI - Tinnitus suppression with threshold and subthreshold sound stimuli. AB - In this preliminary report, we present the results from our investigation of 34 tinnitus patients for tinnitus suppression with frequency-specific sound stimuli within the auditory spectrum. Of this number, 22 (64.7%) experienced suppression, 5 (14.7%) had partial suppression, and 7 (20.6%) were nonresponders. Suppression of peripheral tinnitus may result when mechanosensitive outer hair cells are recruited by sound stimuli that can remain at subthreshold level. The suppression mechanism is possibly explained by the electromodel of the auditory system. This physiological model could be the basis of tinnitus suppression therapy in which a low-intensity, frequency-specific and tinnitus-suppressing sound stimulus is introduced instead of a wide-band masking noise. PMID- 14763324 TI - Audiological evaluation of twenty patients receiving pentoxifylline and prednisone after sudden deafness: prospective study. AB - Idiopathic sudden deafness is defined as sudden sensorineural hearing loss of undetermined etiology. As a consequence, various treatments have been developed for this disorder. Our study evaluated the effectiveness of pentoxifylline and prednisone in such treatment. We analyzed this treatment's results in our patients through conventional audiograms, and speech audiometry was performed in the acute stage and during the treatment. We diagnosed idiopathic sudden hearing loss in 20 patients (8 female and 12 male). The left ear was involved in 9 patients and the right in 11. All patients had been examined by us within 15 days from the onset of hearing loss. We compared the hearing threshold results in the different periods in this prospective study. PMID- 14763325 TI - Balance platform: mathematical modeling for clinical evaluation. AB - This study describes from a technological and mathematical point of view a systematic method for daily patient equilibrium evaluation during clinical work. We present the hardware commonly used for center-of-gravity determination and the basic tasks of the soft ware that runs in a personal computer, including an original procedure for rehabilitation measure. Using a simple approach, the method allows quantification of amelioration of the patient's condition and provides very useful (and, in practice, effective) distribution percent ages for visual, somatosensory, and vestibular contributions to stability or sensory equilibrium organization. PMID- 14763326 TI - Tinnitus outcome profile and tinnitus control. AB - This reports recommends the consideration of development of additional outcome measures to be used as a battery of subjective self-assessment questionnaires for patients with tinnitus. The goal is improved overall care for the tinnitus patient. Five existing outcome measurements have been incorporated into this profile, which is called the tinnitus outcome profile: the tinnitus intensity index, the tinnitus annoyance index, the tinnitus stress test, the tinnitus handicap inventory, and the measurement of depression scale. Frequently, there is an inconsistency between the self-report outcomes and the patient's subjective report to the doctor. This study reports outcomes based on the tinnitus outcome profile for 19 patients who were seen for evaluation and treatment of subjective idiopathic tinnitus of the severe disabling type and who completed a treatment protocol called receptor-targeted therapy directed to the GABAA receptor. The drugs used were gabapentin and clonazepam. Using the 0.05 significance level, the tinnitus intensity index, tinnitus annoyance index, and tinnitus stress test scores all declined significantly over time. In contrast, there was no significant change in either the tinnitus handicap inventory or the measurement of depression scale. The percent of tinnitus control reported to the doctor by the patient did not appear to be correlated with the degree of change measured on the tinnitus outcome profile. PMID- 14763327 TI - Use of high-frequency and muscle vibration in the treatment of tinnitus. AB - Although tinnitus is defined as an internal auditory sensation, external auditory stimuli can mask tinnitus under some circumstances. High-frequency vibration delivered as bone conduction stimulation is effective in masking high-pitched tinnitus. In this preliminary report, somatosensory stimulation in the form of low-frequency muscle vibration can also mask high-frequency tinnitus. Somatosensory stimulation provides fast, immediate relief, whereas high-frequency vibration provides longer-lasting benefit. Either modality can stand alone or can be used in conjunction for tinnitus treatment. A clinically feasible technique has been identified for more wide-scale evaluation. PMID- 14763328 TI - Palatal and middle-ear myoclonus: a cause for objective tinnitus. AB - The cause of objective tinnitus provoked by palatal and middle-ear myoclonus may be identified by magnetic resonance imaging of the central nervous system in the Guilliain-Mollaret triangle or it may be idiopathic. The idiopathic cases are rare. From the literature, one can perceive the following unanswered questions: (1) Are palatal and middle-ear myoclonus different entities or do they usually appear together? (2) Are the clicks being produced by the palatal myoclonus (walls of the eustachian tube slapping together) or by the tensor tympani muscle contractions or by both? (3) Is stapedius muscle myoclonus important in the production of the clicks? (4) What should be the treatment for palatal and middle ear myoclonus? We present two cases of idiopathic palatal and middle-ear myoclonus and discuss these cases in light of the literature and of some ideas of our own. PMID- 14763329 TI - Psychological manifestations of vertigo: a pilot prospective observational study in a Portuguese population. AB - We sought to ascertain the importance of psychological manifestations of vertigo and psychogenic vertigo among a Portuguese population. Sixty patients complaining of vertigo and imbalance were studied over a 2-year period. At each assessment, the patients underwent a general examination, a neurootological evaluation, psychiatric interviews, and psychopathological assessments conducted by a multidisciplinary team. Overall, 38 patients (63.4%) were given diagnoses of some form of psychopathological complaint. A more detailed analysis revealed panic disorder in 9, moderate depressive episode in 42, and mixed anxiety and depressive disorder in 7, whereas the remaining patients (2) suffered from subclinical symptoms of anxiety or depression. Patients with vertigo demonstrate a high incidence of psychopathological complaints, the most common being anxiety disorders. Somatization, obsessive-compulsive behaviors, and depression tend to lessen over time, whereas few changes are seen in anxiety. PMID- 14763330 TI - Electronystagmographic and caloric investigation data about vascular-vestibular dysfunction among patients with vertebrobasilar insufficiency. AB - Audiovestibular symptoms are the first and often the only clinical signs of vertebrobasilar insufficiency. The aim of our investigation was to analyze the results of caloric and electronystagmographic investigation of vascular vestibular disorders among patients with vertebrobasilar insufficiency. We examined a total of 55 men (mean age, 51.8 +/- 8.2 years)--a primary group of 35 men with proven vascular-vestibular dysfunction and vertebrobasilar insufficiency and a control group of 20 clinically healthy men. Standard and electronystagmographic ("Tonnies" type) caloric tests were used to evaluate the human vestibular analyzer. Our data from this investigation show that in both rotatory and caloric tests, the slow-phase speed is the most important index about function of the vestibular analyzer. PMID- 14763331 TI - Bilateral tinnitus due to middle-ear myoclonus. AB - Tinnitus is a common otological symptom. Usually it is subjective (perceived only by the patient); very rarely is it objective (heard by both the patient and the examiner) Objective tinnitus due to middle-ear myoclonus is extremely rare, with only a few case reports published in the literature. We present three cases of objective tinnitus caused by middle-ear myoclonus. All patients were cured by tympanotomy with stapedial and tensor tympani tendon section. PMID- 14763332 TI - Hearing aid fitting: application of telemedicine in audiology. AB - Telemedicine is used as a tool for therapeutic performances with the aid of telematics. The term telematics, a compound of the words telecommunication and informatics, means (in general) electronic transfer of digital data. We describe an application of telemedicine in the field of audiology: the initial fitting, setup, fine tuning, and follow-up fitting of all kinds of digital programmable and digital signal-processing devices and the fitting of hearing aids in real soundscapes. Against a backdrop of limited resources and continuous innovations, telemedicine offers the possibility of increasing the efficiency of audiological methods and decreasing expenses at the same time. PMID- 14763333 TI - Neural response telemetry in cochlear implant users. AB - Programming of multichannel cochlear implants requires subjective responses to a series of sophisticated psychophysical percepts. It is often difficult for cochlear implant patients (especially young prelinguistically deaf children) to provide adequate responses for device fitting. However, the neural response telemetry (NRT) system renders possible the measurement of the compound action potential threshold. We performed NRT examinations in 27 cochlear implant users with Nucleus 24-channel cochlear implants. Measurements were obtained from five electrodes (3, 5, 10, 15, and 20) in each patient. Our goal was to look for correlation between behavioral subjective thresholds and compound action potentials. The action potentials could be elicited in 23 patients in all measured electrodes. The NRT threshold values were highly correlated with electrical threshold levels obtained through subjective responses. Our results suggest that the electrically elicited neural responses may yield very important information for device fitting in patients with cochlear implants. PMID- 14763334 TI - [How should an immunodeficiency be explored?]. AB - When immunodeficiency is suspected, simple investigations allow to direct the diagnosis. Blood cell count brings essential elements: the lymphopenia, signing a deficit of cellular immunity, is often ignored because of the physiological hyperlymphocytosis of the young child. Only a neutropenia lower than 500/mm3 could be responsible for obvious clinical signs. The level of the serum immunoglobulins IgG, IgA, and IgM becomes interpretable only after the age of 4-5 months. In most of the cases, all the elements brought by these simple laboratory investigations jointly to those brought by the medical history and the clinical examination make it possible to target the more specific investigations required in a second time, according to the type of evoked immunodeficiency. PMID- 14763335 TI - [New hereditary immunodeficiencies and genetic predisposition to infective diseases in children]. AB - Since 10 years severe pediatric infections which were idiopathic have now molecular explanation, because new primary immunodeficiencies responsible of these severe infections were identified. These children presented a new kind of hereditary immunodeficiency with severe and/or recurrent infections caused by only one microorganisms family, in opposition to other patients with "classic" primary immunodeficiency. Standard immunologic explorations for example white blood counts, lymphocyte counts, vaccine serology, immunoglobulin levels and complement were normal. However, these children presented a vulnerability, sometimes lethal, caused by one type of microorganism. The aim of this review is to describe 3 new syndromes with a genetic predisposition of infectious diseases: IL-12-IFN gamma axis deficiency (Mendelian susceptibility to mycobacterial disease), STAT1 deficiency (predisposition to viral disease) and NEMO and IRAK-4 deficiencies (predisposition to infections caused by pyogenic bacteria). PMID- 14763336 TI - [Hemophagocytic syndromes and primary immunodeficiencies]. AB - Lymphohistiocytic activation syndromes are characterized by activation and proliferation of T cells and macrophages usually reflecting an inappropriate response of the host to an infection. The clinical manifestations are often alarming symptoms suggestive of severe sepsis. Most patients have a known underlying disease (hemopathy, lupus, Still's disease, HIV infection, bone marrow or organ transplantation). In the few cases that occur apparently without any risk factors, investigations should be done to look for a predisposing inherited disease such as familial lymphohistiocytosis or Purtilo syndrome in boys. The treatment relies on immunosuppressive agents followed by bone marrow transplantation, which can provide a definitive cure in genetics forms. PMID- 14763337 TI - [Evaluation of neutropenia in children]. AB - Many circumstances lead to discover a neutropenia in paediatric practice. In most of the cases, it is an acquired, transient neutropenia, related to a viral or a bacterial infection, a malignant haemopathy, or an acquired auto-immune neutropenia, also called benign chronic neutropenia. Constitutional disorder with neutropenia is more exceptional. Many complex genetic diseases include a neutropenia, among which several immunologic disorders that could be easily diagnosed by immunological tests. Other complex genetic diseases include Shwachman-Diamond syndrome, associating an external pancreatic insufficiency with bone and skin abnormalities; Glycogen storage disease type Ib, with metabolic disorder. Primary constitutional neutropenias are limited to very few entities. Kostmann's disease is a permanent isolated neutropenia, usually associated with a bone marrow granulopoeisis blockage; cyclic neutropenia is characterised by recurrent oscillations (every 21 days) of neutrophil count. Elastase 2 gene mutations have been observed in both diseases. Treatment and prevention of severe infections are a major concern in the management of chronic neutropenia and could be achieved by prophylactic antibiotics (like sulfamethoxazole-trimethoprime) and also G-CSF, which is regularly effective, but could facilitate various side effects. Constitutional neutropenias, especially Kostmann's disease and Shwachman Diamond syndrome, are associated with an increased leukemic risk. PMID- 14763338 TI - Moving towards integrated working in health and social care in Scotland: from maze to matrix. AB - The development of integrated working across health and social care has featured strongly in recent policy directives in both England and Scotland. This is part of a wider agenda of partnership and collaboration, with a range of options from the creation of unified structures as in the care trusts in England to localised arrangements for joint working between individual professionals. This article presents a detailed matrix of drivers and barriers to integrated working which has been developed through a number of case studies of community care practice pursued as part of work undertaken for the Joint Future Group of the Scottish Executive. Drivers and barriers in three key areas are highlighted: national policy frameworks, the local planning context, and operational factors. It is anticipated that the matrix should provide a useful framework for the detailed scrutiny and operationalisation of integrated working. PMID- 14763339 TI - Relative professional roles in antenatal care: results of a survey in Scottish rural general practice. AB - There is evidence of variation and some ambiguity about self-perceived relative professional roles in antenatal care in the UK. There is little information about models of antenatal care provision in UK rural areas. In rural areas, in particular, women have limited choice in accessing health care professionals or alternative primary care delivery settings. In the light of a recent review of Scottish maternity services, it is important and timely to examine models of care and interprofessional working in antenatal care in rural areas. This study explores midwives' and GPs' perceptions about their relative professional roles in remote and rural general practice in Scotland. A questionnaire survey involving all 174 Scottish remote and rural general practices (using one definition of rurality) was conducted, followed by 20 interviews. At least one professional returned a completed questionnaire from 91% of rural practices. A number of areas of dissonance were noted between GPs' and midwives' perceptions of their roles in maternity care and, given the context of service provision, these may impact upon rural patients. Findings are relevant to wider debates on extending the primary care team and strengthening inter-disciplinary working, particularly in rural areas. PMID- 14763340 TI - Building empowering partnerships for interprofessional care. AB - While partnership approaches have the potential to achieve cost-effective quality health care, several attributes of the current context make partnerships difficult to achieve. This paper provides an analysis of the socio-cultural, structural and human challenges to building partnerships at both personal and organizational levels, together with an empowering interdisciplinary approach for overcoming these barriers. Premised on empirical evidence, 'flexible client driven care', currently being tested in the home care sector in Canada, encompasses structures and processes that promote relationship-building and conscientious critical application of individual and collective potential for achieving health care. Strategies for implementing empowering partnership building at both personal and organizational levels are elaborated, together with the challenges encountered. The practical issues addressed afford insights and ideas for others who may be attempting to achieve similar partnership aims. PMID- 14763341 TI - Anger behaviour among professionals in a Norwegian hospital: antecedents and consequences for interprofessional cooperation. AB - The article is an examination of anger behaviour among professionals in a Norwegian hospital, how it develops in the interaction that takes place between the health professionals and the consequences of anger behaviour on work cooperation. The data comes from an ethnographic study of a Norweigian urban hospital and was gathered through interviews, observations and existing documents that were made available. Anger behaviour, in particular from doctors, is a major stress factor in the work day of nurses and has a negative impact on their work environment and professional cooperation and may even reduce the quality of patient care. To a large extent anger behaviour may be understood as an expression of strained interpersonal relationship where contextual factors serve to lower the threshold for keeping such feelings private. PMID- 14763342 TI - Interprofessional collaboration with young people in residential care: some professional perspectives. AB - The article discusses interprofessional collaboration with young people experiencing psychosocial problems living in residential care in Norway. The professionals involved (n = 23) were social workers, psychologists, teachers, doctors/psychiatrists, unqualified graduates and other staff. The aim was to explore the professionals' contributions and grasp a sense of the wholeness of the collaboration process. A grounded theory approach was applied. During the analysis five categories emerged regarding professionals contributions; knowledge of own and others' agency/service, problem perception, priority, commitment and space for action. Three categories emerged regarding interprofessional interaction; building networks, developing trust and using flexibility. The core category was identified as 'readiness to act'. The findings show an apparent contradiction between health and social policy that encourages the standardisation of services and responding flexibly to the needs of young people for 'tailor made' solutions through access to a range of services. A further finding was extensive use of flexibility and willingness to go beyond boundaries leading to the distinction between routinized and radical coordination. PMID- 14763343 TI - In-service interprofessional education improves inpatient care and patient satisfaction. PMID- 14763344 TI - Evaluation of a multi-professional mentoring scheme in primary health care. PMID- 14763345 TI - Evaluation of the Joint Universities Multiprofessional Programme. PMID- 14763346 TI - Experiences of clinical education: a Swedish perspective. PMID- 14763347 TI - [NSAIDs and digestive complications]. PMID- 14763348 TI - [NSAIDs digestive risk: epidemiologic data]. AB - A HIGH RISK: Conventional non-steroidal anti-inflammatory drugs (NSAIDs) produce, by a factor of about 4, a major increase of the secondary GI adverse effect risk (ulcer and bleeding). According to published data, the number of patients to be treated with an NSAID for a duration of more than 2 months, a duration which can result in death, is 1220 (909 to 2500). A LONG TERM RISK: The GI risk induced by an NSAID appears constant over time. Upon discontinuation of treatment with an NSAID, an elevated risk is observed during the first 3 months following the end of treatment. PMID- 14763349 TI - [Risk factors for gastrointestinal bleeding associated with low-dose aspirin]. AB - INTRODUCTION: Low-dose aspirin is the cause of gastro-intestinal and bleeding complications. Different factors may increase the gastro-intestinal (GI) toxicity associated with aspirin. The aim of this review is to attempt to describe these factors. METHODOLOGY: This review is based on an analysis of 24 randomized GI, cardiovascular and neurological studies and 8 case-control and cohort, follow-up studies evaluating GI and bleeding risk factors associated with the ingestion of aspirin at doses of less than 500 mg/day. These studies have been selected based on specific criteria from all works published from 1983 to 2003 (PubMed). RESULTS: The GI toxicity of low-dose aspirin appears to be dose-dependent, apparently starting with 10 mg/day, but this observation has not been confirmed by all studies. The "protective" effect of gastro-resistant coating is debatable. The GI risk is enhanced in the elderly and in particular appears related to concomitant disorders, specifically cardiovascular and neurological disorders, but few studies make it possible to confirm this. A previous history of ulcer or GI bleeding are identified risk factors for GI bleeding, but few studies have been conducted on patients treated with low-dose aspirin to confirm this. Other platelet-inhibiting and anti-coagulant agents potentiate the gastro-intestinal lesional and bleeding risk associated with low-dose aspirin. GI complications are more frequent if aspirin is given in combination with conventional NSAIDs or steroids. In the case of concomitant prescription of low-dose aspirin and an NSAID, the use of a coxib may be considered, but studies are still necessary to evaluate the usefulness of their combination use with low-dose aspirin. In the future, other better tolerated NSAIDs (NO releasing NSAIDs, zwitterion complexes) will be developed. Ongoing studies evaluate the protective risk of misoprostol and proton pump inhibitors in patients at risk for a GI adverse effects, receiving low-dose aspirin. CONCLUSION: The GI risk induced by low-dose aspirin exists and may be increased by different factors related, first to the patient, and second to certain concomitant treatments. Patients at risk must be screened in order to plan a prophylactic strategy. However to date, no compound has been registered for prophylactic treatment of gastro-intestinal risk induced by low dose aspirin. PMID- 14763351 TI - [Is it possible to rate digestive toxicities of NSAIDs?]. AB - EPIDEMIOLOGICAL STUDIES: Most often case-control studies, based on serious GI complications reported in hospitalization statistics are available to compare the gastro-intestinal (GI) safety and tolerability of the various NSAIDs currently on the market. Based on pooled data regarding 5 medicinal products among the most widely used and best documented (ibuprofen, diclofenac, naproxen, indomethacin and piroxicam), the relative result consistency of these studies is striking, thus these 5 NSAIDs may be legitimately classified from the least gastro-toxic (ibuprofen) to the most gastrotoxic (piroxicam). MISSING DATA: But epidemiological survey results may be questioned. Indeed, the volume of prescriptions written, individual dosages and the route of administration are not well documented. Above all, the disease justifying the prescription of an NSAID is never specified. Yet, it is likely that some major anti-inflammatory agents are prescribed more often, at higher doses in patients more seriously and chronically affected, and thus more likely to develop serious gastro-intestinal adverse effects. Data for recently marketed NSAIDs are less comprehensive, with the number of subjects exposed to these new NSAIDs being low, and the validity of the results reported, limited. Regarding currently marketed specific cyclo oxygenase 2 inhibitors, recent cohort studies demonstrating that the relative risk of ulcer is about half compared to subjects treated with traditional NSAIDs are available. IN SUMMARY: The epidemiological survey results can only be extrapolated to individual clinical practice with great caution. PMID- 14763350 TI - [Digestive and hemorrhage complications of low-dose aspirin]. AB - INTRODUCTION: The gastro-intestinal (GI) toxicity associated with high dose aspirin has been fully demonstrated, but remains poorly elucidated at low doses i.e., less than 500 mg/day. Such toxicity is relatively difficult to study because lesional and/or bleeding GI complications are not always well described in studies. The objective of this review is to compile a documented inventory of GI complications induced by low-dose aspirin. METHODOLOGY: This review is based on a detailed review of randomized studies, case-control and cohort studies which aim to study the GI toxicity of low-dose aspirin. These studies have been selected based on specific criteria from works published from 1983 to 2003 (PubMed). RESULTS: In 8 randomized, placebo-controlled or group-controlled studies, erosions, in particular gastric erosions, are more frequent in the elderly than in younger subjects. On the other hand, the prevalence of gastric and duodenal ulcers does not appear to be significantly increased except in specific cases. The most significant complications are GI bleeding described only in 3% of cases. Among randomized studies which have evaluated the cardiovascular and neurological usefulness of low-dose aspirin, 16 have evaluated GI safety and tolerability as a secondary end-point. These studies demonstrated a non significant increase in esophageal, gastric and duodenal ulcers during treatment with low-dose aspirin (Odds Ratio = 1.22, P = 0.08), but a significant increase in bleeding ulcers (Odds Ratio = 1.77, P = 0.04). These complications seem to occur in rare cases (less than 3% of patients) and often seem minor. Nevertheless, ulcers have been less studied than bleeding because few endoscopic analyses have been performed in these studies. The gastro-duodenal bleeding related to erosions or ulcers are significantly more frequent with Odds Ratios between 1.3 and 3.3 (P < 0.05). There are 8 case-controlled studies and 1 cohort follow-up study which confirm the increased GI risk with low-dose aspirin. GI bleeding adverse effects related mainly to gastro-duodenal ulcers are more frequent in case of the regular use of aspirin. CONCLUSION: The GI risk exists, starting with the lowest doses and appears to be dose-dependent. The lesional complications consist mainly of erosive lesions, most often gastric, and rarely true ulcers. Cases of bleeding appear more frequent, but generally are minor. This risk should be taken into account by the prescribing physician and the patient should be informed when treatment with low-dose aspirin is initiated. PMID- 14763352 TI - [What treatments can reduce the digestive complications of NSAIDs]. AB - OBJECTIVE: The aim is to compile a systematic review of literature data on the efficacy of GI protective agents in the prevention of upper GI lesions in a subject receiving long-term therapy with an NSAID. METHOD: We have selected solely experimental studies which have received a very high score in terms of methodology, whether in internal validity or clinical significance. Thus, 35 randomized, controlled studies have been reviewed. RESULTS: Only one large long term study examined the prevention of serious GI risks and misoprostol (800 micrograms/day) showed that it was capable of reducing the incidence of ulcer complications (perforations and bleeding) by 40% compared to placebo. No such data are available for PPI and H2-receptor antagonists (H2-RA). In summary Controlled endoscopic study results for patients with joint diseases receiving NSAID therapy for a period greater than 3 weeks demonstrated that misoprostol, PPI and H2-RA at double-dose are effective in reducing the risk both of gastric and duodenal ulcer. The standard doses of H2-RA are not effective in reducing the risk of gastric ulcer. There are very few studies comparing two gastro-intestinal protective agents and they seem to indicate that misoprostol and proton pump inhibitors are comparable in reducing the risk of gastric ulcer and are better than standard doses of H2-RA and that omeprazol is better than misoprostol at reducing the risk of duodenal ulcer. Tolerability of PPI and H2-RA are appreciably better than misoprostol which induces intestinal disorders. PMID- 14763353 TI - [Prescription of NSAIDs in patients treatment with platelet inhibitors or anticoagulants]. AB - PLATELET INHIBITION: Conventional NSAIDs and aspirin have platelet-inhibiting properties via platelet cyclo-oxygenase-1 (Cox-1) isoenzyme inhibition. Among the NSAIDs which induce reversible inhibition of Cox-1, naproxen seems to have a potent platelet-inhibiting action associated with a decrease in cardiovascular clinical events in secondary prevention. Flurbiprofen is indicated in the secondary prevention after myocardial infarction. Conversely, no conventional NSAID is indicated and none have demonstrated efficacy in primary prevention. GI BLEEDING: The platelet-inhibiting action of conventional NSAIDs and aspirin is a key factor in explaining the increased risk of bleeding reported with these medicinal products. Digestive system bleeding is the main risk. Few data are available regarding bleeding other than gastro-intestinal. COX-2 SPECIFIC INHIBITION PROPERTIES: Conversely, Cox-2 specific inhibitors have no platelet inhibiting properties: the pharmacological profile of Cox-2 SI is therefore different from that of conventional NSAIDs. These medicinal products are anti inflammatory, analgesic, platelet-inhibiting agents which are not harmful to the GI mucosa and which probably induce less bronchospasms. In patients with cardiovascular risks, Cox-2 SI can and should be used in combination with platelet-inhibiting agents, in particular aspirin, because they do not alter the aspirin effect on platelets, contrary to what is observed with conventional NSAIDs. PRESCRIPTION WITH ANTI-COAGULANTS: The concomitant prescription of conventional NSAIDs with anti-coagulants is not recommended because, in addition to the possible potentiation of oral anti-coagulants, NSAIDs increase the risk of serious bleeding adverse events due to their irritative effect on the GI mucosa and their platelet-inhibiting effect. This risk might be lower with Cox-2 specific inhibitors. PMID- 14763354 TI - [Justification and indications for coxib therapy combined with gastro-protective agents]. AB - IN THEORY: Combination use of a coxib with a gastro-protective agent does not appear justified since results of studies conducted on patients of all types demonstrated that the upper-digestive tract safety and tolerability profile of coxibs is good. IN SUMMARY: On the other hand, prescription-related descriptive data demonstrate that in real-life situations such a combination occurs in approximately 25% of cases. Two cases should be differentiated: in about half of cases, patients are receiving long-term therapy with a proton pump inhibitor (PPI) (for reflux oesophagitis, oesophagitis or dyspepsia) and when an NSAID is necessary, the prescribing physician logically turns to a coxib; in the other half of cases, it involves a true simultaneous co-prescription of a coxib and a gastro-protective agent (most often by far a PPI). Thus, it is only in 10-15% of patients that prescription of a coxib leads to prescription of a PPI. A SAFETY ATTITUDE: This approach can be justified by the physician's desire to be as cautious as possible in dealing a patient at high risk of GI disorders (elderly patients, previous history of digestive bleeding, co-prescription of aspirin and/or a steroid, co-morbidity) or in patients for whom the occurrence of GI bleeding would be very serious (i.e. a patient in poor condition due to multiple disorders, treatment with an oral anti-coagulant or heparin). In both cases, if the prescription of an NSAID cannot be avoided, it is therefore logical first to choose a coxib and second to prescribe simultaneously a gastro-protective agent. PMID- 14763355 TI - [Physician and patient knowledge of NSAID digestive complication risks]. AB - REQUIREMENT: Information given to the patient is a topic which is especially of current interest, and which has been taken into account in the recent law of March 4, 2002 on patient's rights. But what about non-steroidal anti-inflammatory agents, that is, one of the most widely prescribed classes of medicinal products worldwide and which induce the highest number of sometimes fatal adverse effects (adverse drug reactions)? FREQUENT MISREADING: The aim of this update is thus to review once again the rare studies published on this topic, a review necessary prior to the formulation of recommendations regarding information to be given to the patient before prescribing him/her an NSAIDs. A review of the literature illustrates the common misunderstanding that patients have (nearly 1/4 to 1/2) regarding the potential serious gastro-intestinal adverse effects of NSAIDs. RECOMMENDATIONS: We propose that the subjects at the very least, be informed first of the risk of dyspepsia, which is the most common side-effect but usually not predictive of a serious adverse effect; then we would point out the risk of gastro-duodenal ulcer whose complications certainly are rare but can be fatal; and lastly the existence of prophylactic strategies, including the recognition of main risk factors and some medical options, such as use of synthetic prostaglandins, proton pump inhibitors or the preferential use of coxibs. PMID- 14763356 TI - [Are dyspepsia or gastroesophageal reflux predictive of serious digestive events during treatment with NSAIDs? How should they be treated?]. AB - The prevalence of dyspepsia is high in the general population. Its incidence varies depending on the study and the definition chosen for dyspepsia. Treatment with NSAIDs increases the risk of dyspepsia. The absence of dyspepsia does not eliminate a potential serious GI event during treatment with an NSAID, and patients with dyspepsia do not have an increased risk of GI complications. What about gastro-oesophageal reflux? Symptoms of gastro-oesophageal reflux are frequent during treatment with an NSAID, but the NSAID-related gastro-oesophageal reflux-inducing effect has not been demonstrated. On the contrary, this causal relationship has been established in the occurrence and worsening of lesions of oesophagitis. From the standpoint of management, the existence of dyspepsia or gastro-oesophageal reflux before prescription of an NSAID should be treated and investigated if necessary (patient older than 50). In the event of chronic gastro oesophageal reflux, endoscopy is necessary to screen for and treat oesophagitis before initiating treatment with NSAID. In case of dyspepsia or gastro oesophageal reflux developing during treatment with an NSAID, endoscopy is not a systematic procedure to be performed in a first phase. It is indicated in case of non-response to a symptomatic therapy and in case of a GI lesion risk (patient older than 50). PMID- 14763357 TI - [Risk factors for upper digestive tract toxicity with NSAIDs (excluding Helicobacter pylori)]. AB - AN ABSOLUTELY CERTAIN RISK: The non-selective non-steroidal anti-inflammatory drugs increase the risk of gastro-duodenal ulcer by 3 to 5 and the incidence of the symptomatic ulcers is 1 per 100 patients treated per year History before anti Cox-2. HISTORY BEFORE ANTI COX-2: The past history of ulcer increases the risk of relapse by 14 to 17 with non-steroidal anti-inflammatory drugs intake compared to patients without previous ulcer. The severity of the ulcers induced by non steroidal anti-inflammatory drugs increases with the age. The ulcer risk is constant with the time and thus the prevalence increases with the duration of the treatment. The concomitant treatments by anticoagulants or aspirin increase the risk of digestive events by 2 to 3. A heavy consumption of alcohol is a factor of risk. Smoking is a minor factor. WITH ANTI COX-2: The incidence of the symptomatic ulcers is reduced by 60% among patients treated by Cox-2 specific antagonists. This reduction is identical that the patients have or not risk factors, and thus, the digestive complications are not suppressed when giving non specific antagonists to the patients with risk factors. Their incidence cannot be negligible among patients cumulating at least two factors and treated by Cox-2 specific antagonists (from 5 to 16% per year). The benefit related to the IN SUMMARY: Cox-2 specific antagonists seems dramatically reduced by the concomitant aspirin intake. Among the patients who have a symptomatic ulcer induced by non steroidal anti-inflammatory drugs, 81% did not have previous symptoms or risk factors. PMID- 14763358 TI - [Role of Helicobacter pylori in drug-induced gastrointestinal bleeding]. AB - TWO CO-FACTORS: Helicobacter pylori (Hp) infection and ingestion of NSAIDs are two factors which play a major role in the occurrence of gastric and duodenal ulcers, simple or complicated by perforation and GI bleeding. Hp increases the risk of ulcer in patients receiving an NSAID and the effects of Hp and NSAIDs are additive. Following GI bleeding in a patient receiving a conventional NSAID, the eradication of Hp does not make it possible to protect the patient from another episode of GI bleeding and proves to be a much less effective therapy than continuous therapy with proton pump inhibitors. ROLE OF COX-2 INHIBITORS: COX-2 inhibitors (celecoxib, rofecoxib) are associated with a major decrease in the number of symptomatic and/or complicated ulcers compared to a conventional NSAID. Two randomized studies using doses four times higher than those recommended, have demonstrated that serious GI complications associated with NSAIDs (perforation, GI bleeding) were decreased by half compared to those by use of a conventional NSAID at the usual therapeutic dosage. Considering the patient's previous history of GI disorders prior to initiation of treatment and Hp(+) serology, it appears that decreasing risk of symptomatic or complicated gastro-duodenal ulcer was higher in treatment with Rofexocib in patients who did not have a previous history of ulcer, whatever their Hp status. On the other hand, in the group with a history of GI disorders, the decrease in the risk of ulcer was lower than in patients who were Hp(+). ROLE OF ASPIRIN: The causal relationship between intake of aspirin and Hp appear to be more complex. Thus, it appears that eradication of Hp in patients who presented with GI bleeding and scheduled to resume low-dose aspirin had the same efficacy as a daily dose of 20 mg of omeprazole for a 6 month period. PMID- 14763360 TI - Pathophysiology and diagnosis of asthma. AB - The process of asthma includes swelling, bronchoconstriction, and excess mucus production driven by the redundant nature of the inflammatory process. Untreated or undertreated, this process has the potential to cause airway remodeling and the permanent loss of pulmonary function. It is essential that all health care professionals involved in the care of patients with asthma have as their major goal the treatment and prevention of the inflammatory process. PMID- 14763361 TI - Pulmonary function testing in asthma: nursing applications. AB - Spirometry has become the most widely used assessment of pulmonary function for diagnostic and prognostic purposes. This article reviews the indications for spirometry in persons who have asthma, the parameters measured, acceptable testing techniques, acceptability, quality assurance criteria, and basic interpretation of results in evaluating the person who has asthma. Understanding basic spirometry is an invaluable aid to the nurse and nurse practitioner who provide care to children and adults who have asthma. PMID- 14763362 TI - General principles of asthma management: symptom monitoring. AB - The first step in optimal asthma care is identifying and properly diagnosing patients who have asthma then classifying asthma severity carefully and accurately to ensure proper treatment. Objective monitoring of pulmonary function using spirometry and peak flow monitoring, subjective assessment using symptom identification, and physicians' acceptance of National Asthma Education Program's Expert Panel guidelines are needed to diminish the consequences of undertreatment of asthma. Persistent asthma requires continuous long-term controller therapy. Erratic and insufficient use of medication must be addressed. Clinicians should encourage patients to manage their asthma using routine peak flow monitoring and symptom assessment then intervene according to their asthma action plan. These efforts allow patients to take charge of their asthma instead of the asthma taking charge of them. PMID- 14763363 TI - General principles of asthma management: pharmacotherapy. AB - Asthma, a disease of urbanized societies, has been increasing at an alarming rate on a worldwide basis. In recognition of this problem, the NHLBI developed the NAEPP in 1991. An expert panel was appointed that developed guidelines for the diagnosis and management of asthma. The original report has been updated several times, most recently in 2002. In this article the author focuses on the general principles of asthma pharmacotherapy in accordance with the conceptual framework that characterizes drugs as relievers or controllers. The preeminent use of ICS in the management of asthma sufferers who are characterized as having more than mild intermittent disease is emphasized, as is the principle of matching therapy with severity of disease. PMID- 14763364 TI - General principles of asthma management: environmental control. AB - Environmental control is one of the major goals of asthma management. House dust mite environmental control and the reduction of other indoor environmental allergen triggers, such as animal dander, cockroaches, and mold spores, are important for individuals with asthma. Patients who have sensitivities to the allergens they produce should receive advice on environmental control reduction measures to prevent asthma symptoms. Nurses are in a key position to make proper environmental control recommendations to patients. PMID- 14763365 TI - General principals of management: education. AB - Asthma education is an essential part of the treatment of this disease. Health care professionals must establish a partnership with the patient with asthma and the patient's family to devise a plan of care with which the patient voluntarily will comply. The partnership with the patient begins at the first encounter and continues throughout the therapeutic relationship. Each member of the health care team can be instrumental in reinforcing the crucial information the patient must know to be an informed participant in his or her care. Nursing professionals are in a particularly advantageous position to foster this partnership because of their patient-focused outlook and the quality of time spent with patients. When the partnership is based on mutual trust and cooperation, the clinician can direct asthma care that is consistent with current expert guidelines. Educational interventions should be meaningful to the patient, learner centered to incorporate the patient's needs, and sensitive to the patient's cultural influences. The patient and his or her significant social and family support should be actively involved. The clinician should be alert to the patient's readiness to learn and tailor the message to suit the setting in which it is delivered. The patient should receive information that allows his or her participation in goal setting for treatment. Essentials to be included are the significance of the diagnosis, basics about inflammation as the primary cause of symptoms, the difference between controllers and relievers, how to use the medications and monitoring devices, how to reach the provider, and the need for continuous ongoing interaction with the clinician. Goals set in the partnership are objectified in the asthma action plan or guided self-management plan. The success of the treatment can be assessed from the patient's improved asthma control and reduced reliance on emergency treatment. Every health care provider is a potential wealth of patient education. Every patient encounter is an opportunity to reinforce knowledge and proficiency in asthma management. Nursing professionals can play a fundamental and crucial role in asthma education by maintaining the focus of the medical treatment on the priorities in asthma care- the learning needs and goals of the patient. PMID- 14763366 TI - Asthma management across the life span: the child with asthma. AB - Childhood asthma has an adverse impact on children, families, and society. Treatment of asthma presents special challenges related to diagnosis, ongoing symptom monitoring, and treatment when the patient is a child. To be effective, treatment needs to be medically sound and developmentally appropriate. PMID- 14763367 TI - Asthma education: special applications for the school-age child. AB - School-age children have learning needs unique to their psychosocial and cognitive developmental stage. Teaching that requires the learner to master abstract concepts is particularly challenging. This article describes how the idea for an interactive doll/model evolved among a group of health care professionals at a day camp for children with asthma. How this group of educators designed, developed, and field-tested the model in clinic and classroom environments are presented and discussed. Strategies related to a class on key concepts of asthma education illustrate the various ways the model can be used to facilitate learning. Field testing has shown that this teaching aid enhances knowledge retention and has a high degree of learner satisfaction. PMID- 14763368 TI - Asthma management across the life span: the childbearing woman with asthma. AB - Asthma can pose a serious threat to the pregnant mother and her fetus if not treated appropriately. Studies analyzing the causes of death in severe asthma have shown that most occur outside of the hospital with neither the physician nor the patient appreciating the seriousness of the symptoms [31]. For this reason, a pregnant woman with a diagnosis of asthma must be evaluated thoroughly and managed aggressively by an obstetrician and pulmonary specialist or a maternal fetal medicine specialist trained in high-risk pregnancies. Any evaluation or assessment of the pregnant asthmatic must be interpreted in light of pregnancy induced changes. Treatment must be focused on set goals to correct maternal hypoxia, relieve bronchospasm, ensure adequate ventilation, and optimize uteroplacental exchange preventing fetal hypoxia. PMID- 14763369 TI - Asthma management across the life span: applications for the adult and older adult. AB - Asthma is one of the most common and chronic diseases of adults and creates substantial health problems. The disease must be diagnosed appropriately, its severity assessed, and treatment prescribed that matches the level of severity. Patient education in self-management techniques and attention to the problems of adherence are essential for long-term management. Comorbid conditions should be suspected and treated when asthma becomes difficult to control. Asthma in the elderly is a challenging but frequent problem that requires particular attention to controlling the causes of excessive morbidity and mortality. All health care professionals have an important role in controlling this common disease. PMID- 14763370 TI - Exercise-induced asthma. AB - EIB is a common manifestation in diagnosed asthmatics and is a sensitive warning sign of worsening asthma. EIB may occur as an isolated entity in the absence of signs and symptoms of chronic asthma. Careful diagnosis is crucial in determining if this condition represents a preasthmatic state or simply a transient condition resulting from individual sensitivity to hyperventilation. The pathophysiology of EIB is hypothesized to result from rapid airway rewarming after hyperventilation. Effective treatment modalities include behavioral techniques and pharmacologic agents. Appropriate treatment regimens consisting of pharmacologic or behavioral modalities should prevent serious sequelae and should enable the patient to participate at all levels of athletics. The nurse is an essential member of the team identifying, preventing, and treating exercise-induced bronchospasm or asthma. PMID- 14763371 TI - Asthma, allergy, and upper airway disease. AB - The practitioner must be aware of the strong interrelationship between asthma, allergy, eczema, otitis media, viral upper respiratory infection, and sinusitis. When allergy, eczema, otitis media (particularly after the age of 2 years), viral upper respiratory infection, or sinusitis is present in a patient, asthma must be considered as possible sequelae. If a patient has symptoms of intermittent or persistent asthma, consideration must be given to the presence of allergy, eczema, otitis media, viral upper respiratory infection, or sinusitis as comorbid conditions or possible triggers for the asthma symptoms. Failure to evaluate the entire upper and lower airway for these interrelated conditions will result in incomplete treatment and incomplete symptom relief [61]. This article reviews the strong association between diseases of the upper and lower airway [62,63]. Treatment and control of upper airway symptoms is essential for control of asthma symptoms. Control of allergy, eczema, otitis, sinusitis, and viral symptoms result in improved outcomes for asthma patients. Reduction of these asthma triggers and comorbid conditions is also likely to reduce asthma medication requirements while improving symptoms status. PMID- 14763372 TI - Management of asthma emergencies. AB - The patient with asthma experiencing an acute exacerbation has been described clinically via case scenarios. Exacerbation severity has been defined, and the specific factors that differentiate the exacerbation have been identified. Home and hospital management has been delineated, and algorithms identifying pattern recognition and management that assist in clarifying the standard for care of an exacerbation have been provided. The nurse's role in asthma care includes administering therapy according to evidenced-based guidelines and providing targeted education to the patient and family that results in trigger avoidance and control of asthma. PMID- 14763373 TI - Asthma epidemiology: risk factors, case finding, and the role of asthma coalitions. AB - Asthma has no cure, and given that clinicians still do not understand what causes it in the first place, it is not preventable. With implementation of environmental control measures to minimize exposure to allergens and irritants, appropriate pharmacologic therapy, and patient and health care provider education, the disease can be managed successfully. Health care access and appropriate case management are essential components of any plan to address asthma as a public health problem. Asthma is now one of the most important diseases of childhood in developed countries. In the International Study of Asthma and Allergies in Childhood (ISAAC) study, the highest asthma prevalence was observed in westernized English-speaking countries (e.g., the United Kingdom, Australia, and New Zealand), with much lower prevalence rates in Eastern Europe, India, China, other countries in Asia, and Africa [18]. This observation has led to the belief that the rapid increases in asthma prevalence are more likely to be attributable to environmental than genetic factors. The rising prevalence in the United States and worldwide seems to be correlated with modern industrialization, suggesting that changes in the ambient environment may contribute to this increase in morbidity and mortality. Indoor factors that have been suggested to influence asthma pathogenesis include allergy to house dust mite and cockroaches, poor heating and ventilation, and exposure to cigarette smoke and other irritants. Scientific evaluation of the links between indoor and outdoor air pollution and asthma is incomplete, however. Much work remains to be done in defining the environmental factors that may cause asthma and that may trigger asthma exacerbations in individuals with the disease. Until then, clinicians must focus on developing a hard-hitting, comprehensive, interdisciplinary asthma program and work toward its successful and broad-based implementation. It is the only way to win the "war against asthma." PMID- 14763374 TI - Symptom perception and respiratory sensation: clinical applications. AB - This article has described symptom perception and its relation to asthma management. Underestimation of airway obstruction in persons with asthma is a common and serious problem that is linked to fatalities in persons with asthma. Strategies to identify and manage the poor perceiver have been suggested. PMID- 14763375 TI - Living with asthma. AB - Living with asthma is a challenging task. Learning to live with asthma using a self-management plan helps patients to achieve control of their asthma. Self management and control tend to improve quality of life. This article relates one nurse's experience of living with asthma to self-management, partnership with the health care team, quality of life, and control of asthma. PMID- 14763376 TI - [Urology imaging: intravenous urography]. PMID- 14763377 TI - [Urology imaging: renal arteriography]. PMID- 14763378 TI - [Urology imaging: lymphography-cavography]. PMID- 14763379 TI - [Urology imaging: RPU-antegrade opacification-MRUC]. PMID- 14763380 TI - [Urology imaging: ultrasonography]. PMID- 14763381 TI - [Urology imaging: tomodensitometry]. PMID- 14763382 TI - [Urology imaging: MRI of the urinary tract]. PMID- 14763383 TI - [Urology imaging: positron emission imaging]. PMID- 14763384 TI - [Urology imaging: interventional imaging in urology]. PMID- 14763385 TI - [Urology imaging: risk of exposure to X rays]. PMID- 14763386 TI - [Urology imaging: iodinated contrast agents]. PMID- 14763387 TI - [Urology imaging: contribution of imaging in prostatic cancer]. PMID- 14763388 TI - [Urology imaging: renal cancer]. PMID- 14763389 TI - [Urology imaging: imaging of adrenal incidentaloma]. PMID- 14763390 TI - [Urology imaging: contribution of imaging in upper urinary tract tumors]. PMID- 14763391 TI - [Urology imaging: contribution of imaging in bladder tumors]. PMID- 14763392 TI - [Urology imaging: imaging of germ cell tumors of the testis]. PMID- 14763393 TI - [Urology imaging: contribution of imaging in penile tumors]. PMID- 14763394 TI - [Urology imaging: imaging and urinary calculi]. PMID- 14763396 TI - [Urology imaging: contribution of imaging in benign prostatic hypertrophy]. PMID- 14763395 TI - [Urology imaging: contribution of imaging in urinary tract infections]. PMID- 14763397 TI - [Urology imaging: imaging of vascular ports for hemodialysis]. PMID- 14763398 TI - [Urology imaging: pre-transplantation assessment]. PMID- 14763399 TI - [Urology imaging: contribution of imaging in kidney transplantation]. PMID- 14763400 TI - [Urology imaging: imaging of female pelvic prolapse]. PMID- 14763401 TI - [Urology imaging: contribution of medical imaging in renal and ureteral injuries]. PMID- 14763402 TI - [Urology imaging: contribution of imaging in bladder injuries]. PMID- 14763403 TI - [Urology imaging: radiologic evaluation of urethral injuries]. PMID- 14763404 TI - [Urology imaging: testicular and penile injuries]. PMID- 14763405 TI - [Urology imaging: imaging and erectile dysfunction]. PMID- 14763406 TI - [Urology imaging: imaging and Peyronie's disease]. PMID- 14763407 TI - [Urology imaging: integral digital environment in urology. PACS]. PMID- 14763408 TI - Approach to arthritis: clinical guideline 2003. PMID- 14763409 TI - Hyperuricaemia and gout: clinical guideline 2003. PMID- 14763410 TI - Osteoarthritis: clinical guideline 2003. PMID- 14763411 TI - Rheumatoid arthritis: clinical guideline 2003. PMID- 14763412 TI - [Patients with terminal uro-oncologic diseases]. PMID- 14763413 TI - [Penile fracture]. AB - OBJECTIVES: Rupture of the corpora cavernosum is an uncommon pathology requiring immediate treatment. We performed a bibliographic search, and analyzed different diagnostic and therapeutic methods. METHODS/RESULTS: Two cases of traumatic rupture of the corpus cavernosum are described. The clinical features, penile ultrasound findings and immediate surgical treatment are discussed. CONCLUSIONS: Penile fracture is an uncommon pathology. Generally, history and physical examination are enough to establish the diagnosis. Penile ultrasound can help in case of doubt. Most urologists deem immediate surgery the standard therapy. PMID- 14763414 TI - [Iatrogenic ureteral injury]. AB - OBJECTIVES: To determine the most frequent causes of ureteral injury, and the way to correct severe damage of the ureter. METHODS: We reviewed the medical records of 25 patients who suffered ureteral injuries during abdominal or pelvic surgery in a five-year period; endoscopic injuries were excluded. RESULTS: The most common presentation included pain, urinary incontinence and fever. Hysterectomy was the most frequent operation causing ureteral injuries. The diagnosis was made by intravenous urography and ultrasound. Ureteral reimplantation using a Boari's flap, without other complications, was the preferred treatment due to ureteral damage. Right side and distal third of the ureter where the most frequently affected sites. CONCLUSIONS: In cases of iatrogenic injuries of the distal third of the ureter with severe lesions, ureteral reimplantation with a Boari's flap may be the option. PMID- 14763415 TI - [Technical principles of prostatic cryosurgery (Part 1)]. AB - OBJECTIVES: Cryosurgery is defined as in situ freezing of tissues. Criosurgery in the treatment of prostate cancer aims to: 1- Eliminate all the glandular tissues and completely destroy the adenocarcinoma, 2- With a minimum number of complications. a) Preserving neighbour structures: bladder neck, striated sphincter, and rectum. b) Preserving urethral mucosa to reduce the elimination of sloughing tissues. METHODS: We describe the technique of prostatic cryosurgery including all innovations introduced during the last ten years. The map with the number and situation of the 6-8 crioprobes should be done in such a way that the iceball created around each of them overlaps the contiguous ones. Probe-mapping and icing strategy together get the prostate included in a unique, solid mass, in which temperature descents homogeneously and spreads out of its limits, so that intraprostatic and extraprostatic temperatures (sensors at the apex, Denonvilliers fascia and both neurovascular bundles) are below -40 degrees C. RESULTS: The standardized technique is based on: Substitution of liquid nitrogen by argon. Use of 6 to 8 cryoprobes. The probes are properly inserted into the perineum percutaneously under transrectal ultrasound guidance. Ice ball evolution control by biplane transrectal ultrasound. Separation of the two layers of Denonvilliers' fascia by injection of saline solution. Real-time measurement of temperature descent. Minimal temperature lower than -40 degrees C Commercial ureteral heating system. Double cycle of freezing-unfreezing Administration of hormonal therapy during time enough to avoid the gland size to exceed 45-50 cc. CONCLUSIONS: We describe the most advanced technique for prostatic freezing, highlighting the basic requirements to achieve the objectives pursued. It has improved oncologic results, in relation to PSA and percentage of negative biopsies, and has dramatically reduced complications. PMID- 14763416 TI - [Bladder calculi. Is extracorporeal shock wave lithotripsy the first choice treatment?]. AB - OBJECTIVES: To evaluate the effectiveness of extracorporeal shock wave lithotripsy (ESWL) treatment of bladder stones using a retrospective study. METHODS: Between February 1991 and June 2002, 45 patients with bladder stones were treated (41 males and 4 females). Age ranged from 23 to 87 years. 63% had previous renal-ureteral lithiasis and 29% had undergone upper urinary tract ESWL. We used the Storz Modulith SL 10/SL 20. Treatments were performed using intravenous analgesia on an outpatient basis. RESULTS: 83 sessions of ESWL were performed on 53 stones. Complete fragmentation and elimination was achieved in 55% of the patients after a single session, 26.7% of the patients after 2 sessions, 6.7% after 3 sessions, 8.8% four, and 2.2% five. Mean number of shock waves was 3196.3 with an average 7-8 Kv. 8.5% had stone recurrence due to residual lithiasis, whereas 79% achieved total elimination. 13% required endoscopic procedures to evacuate stone fragments impacted in urethra. 6.6% required transurethral prostatic resection after ESWL. CONCLUSIONS: ESWL therapy is an effective option for the treatment of patients with bladder stones, non invasive, with low morbidity, without need for anesthesia, and outpatient. The effectiveness is high (79% of the patients stone free) and even higher when treating smaller stones. PMID- 14763417 TI - [Measuring the degree of patient satisfaction treated at an extracorporeal lithotripsy unit]. AB - OBJECTIVES: To determine by questionnaires the level of satisfaction of patients diagnosed of urinary lithiasis treated by extracorporeal shock wave lithotripsy (ESWL) in our Renal Stone Centre. METHODS: A total of 100 patients with urinary lithiasis who underwent ESWL, 54 males and 46 females with an average age of 50 years, answered a Lickert type questionnaire (score 1 to 5) consisting of 6 questions to evaluate the patient satisfaction regarding the treatment received. The relationship between answers to each question and patient's clinical characteristics were analyzed applying chi-square and linear regression to determine the level of significance of these relationships. RESULTS: General satisfaction level was high, with a mean score of 4.33 for the six questions. The most valued question was the one referred to assistance received in relation to the planned medical one. Questions referred to treatment suitability, expected timing, and absence of complexity were also highly valued. The least valued question was the one referred to pain level during treatment, which level was kept in a good approval degree with a mean score of 3.81 despite the fact that some patients said they suffered more pain than expected. CONCLUSIONS: ESWL resulted in a high level of satisfaction among patients treated for urinary lithiasis, revealed not only by its suitability, easy planning and performance, but also by its low level of complexity. Pain experienced by these patients during treatment was well-tolerated, so that general use of anesthesia was not required. PMID- 14763418 TI - [Non-obstructive azoospermia: 2 new cases of Sertoli cell only syndrome]. AB - OBJECTIVES: We report 2 cases of Sertoli cell only syndrome and perform a bibliographic review. METHODS: Clinical and pathologic study of 2 male patients 33 and 37-year-old respectively who presented with infertility. RESULTS: Testicular biopsy, the most important diagnostic test in the study of infertile men, showed bilateral Sertoli cell only syndrome. CONCLUSIONS: The Sertoli cell only syndrome is an entity in which seminiferous tubules show a total absence of germ cells with Sertoli cells alone. Etiology is unknown in most cases. Testicular biopsy is performed in these azoospermic men to rule out tubular obstruction or abnormalities of the spermatogenesis. Assisted reproduction techniques are a therapeutic option, making genetic counselling necessary due to the risk of transmission of the genetic defects that cause male infertility. PMID- 14763419 TI - [Iontophoresis in Peyronie's disease. Our experience]. AB - Peyronie's disease is a type of acquired penile incurvation of unknown origin yet, which generates from the difficulty of symmetric expansion of the corpora cavernosum produced by fibrous plaques in the middle line of the albuginea. OBJECTIVES: To define the results of iontophoresis among the wide spectrum of treatments used for this disease. METHODS: 61 patients treated by iontophoresis with orgotein in weekly sessions. RESULTS: Pain improvement 42.6%, incurvation improvement 6.1%, improvement of both 21.3%. CONCLUSIONS: In patients with Peyronie's disease iontophoresis with orgotein diminishes pain with erections, and, to a lesser extent, penile incurvation. PMID- 14763420 TI - [Magnetic resonance urogram as alternative to i.v. urography. Report of 3 cases. Review of the literature]. AB - OBJECTIVES: Intravenous urography (IVU) has been classically considered as an essential examination in the great majority of urologic diseases due to the great amount of information that supplies both functional and morphological, mainly for ureteral abnormalities where other exams do not reach enough definition. Nevertheless, there are cases in which it is not possible to perform an intravenous urography due to allergy to iodine contrasts or renal failure with impossibility of contrast excretion. METHODS: We report three cases which represent the example of the indications of MRI urography as an alternative to IVU in cases in which this latter cannot or should not be performed. RESULTS: MRI urography is completely accepted as a diagnostic technique for urologic diseases as an alternative to IVU. It has advantages, such as obtaining images in multiple planes, avoiding the use of ionizing radiation, that there is no evidence of it causing cell damage, that it does not require glomerular elimination of any substance so it can be used in patients with renal failure and allergy to iodine contrasts. CONCLUSIONS: The use of MRI urography results in an advantage for patients with renal failure, iodine contrasts allergy, or greater risk of contrast nephrotoxicity as it is the case of renal transplant patients. PMID- 14763421 TI - [Primary urothelial carcinoma of the bulbomembranous urethra. Histologic and immunohistochemical study of a case]. AB - OBJECTIVE: We report a case of a primary urothelial carcinoma of the bulbomembranous urethra, with special emphasis on histopathological and immunohistochemical findings. METHODS/RESULTS: A 63-year-old man presented urethral obstruction symptoms. A radical phalectomy was performed and a 4.5 x 4 cm bulbomembranous urethral tumor was observed. Histopathological analysis disclosed an urothelial carcinoma, that showed positive immunostaining for cytokeratin AE1/AE3, cytokeratin 7, carcinoembrionic antigen and epithelial membrane antigen. The patient received radiotherapy and adjuvant chemotherapy and is currently free of disease. CONCLUSION: Posterior male urothelial carcinoma of the urethra is a rare neoplasm that usually is mistaken clinically for a benign lesion. The diagnostic is generally made in advanced stages with a poor therapeutical response. PMID- 14763422 TI - [A chicken bone as cause of Fournier gangrene]. AB - OBJECTIVES: Fournier's gangrene is a synergistic infective necrotizing fascitis, which involves perianal, perineal and genital regions, with rapid evolution and severe prognosis. We perform a literature review and report a recent case. METHODS: We report the case of a male patient who presented with scrotal pain and history of excretion of a chicken bone in a bowel movement. RESULTS: Despite aggressive surgery and wide spectrum antibiotic, the prognosis severe and the mortality is high. CONCLUSIONS: Rapid and accurate diagnosis remains the key to achieving a successful outcome. Early, wide and repeated debridement procedures reduce the mortality. PMID- 14763423 TI - [Neurogenic bladder caused by hemorrhagic necrotizing myelitis]. AB - OBJECTIVES: To report one case of flank pain irradiated to lower extremities, progressive paresthesias and functional difficulty leading to functional impotence of the lower limbs, and difficult voiding. METHODS: Neurologic physical examination showed abolished osteotendinous reflexes, indifferent plantar, L1 hypoesthesia, L3 anesthesia, and lower limbs hypotony. Multiple complementary exams were performed to get the final diagnosis of hemorrhagic necrotizing myelitis by MRI, after differential diagnosis with cavernous haemangioma. We performed a bibliographic search in Pub-Med (MEDLINE) using the terms "bladder, neurogenic (MESH) and myelitis (MESH)". Most publications correspond to case reports and prospective studies of cases of acute myelitis of different etiologies (arteriovenous malformations, multiple sclerosis, neoplasia, sarcoidosis, HIV infection, spinal cord compression,...) reviewing the diagnostic and therapeutic procedures. RESULTS: Neurologic examination, imaging studies and analytical determinations in cerebrospinal fluid (CSF) and blood were conclusive to confirm the level of the lesion and the most probable etiology in both the present case and those provided by the literature. Urodynamic study showed a neurologic bladder both at 4 and 24 months, being this the main sequel in our patient. The clinical repercussion derived from the level of the lesion improved in relation to walking and muscular balance, but the voiding condition persisted with bladder hyperreflexia and dyssynergia. CONCLUSIONS: The clinical picture alerts about spinal involvement. Radiological tests, supported by analysis, allow to identify the level of the lesion, and in many cases the cause. Bladder dysfunction is common in acute myelopathies, and urodynamic studies help to label the picture and manage it. PMID- 14763424 TI - [Multicystic mesothelioma of the testicular tunica vaginalis]. AB - OBJECTIVES: To report an exceptional case of multicystic mesothelioma of the testicular tunica vaginalis. METHODS: A 72-year-old male was referred for study of a scrotal mass. Physical examination, blood tests, and ultrasound were performed prior to surgical excision and pathologic study of the lesion. RESULTS: The ultrasound study showed a multilobar cystic lesion near the spermatic cord. Pathology reported multiple 3-4 mm cystic formations, with rudimentary papillae covered by a hyperchromatic epithelium and vimentin (+), CD 34 (+) immunophenotype. CONCLUSIONS: Multicystic mesothelioma is a rare form of mesothelioma, easy to recognize but infrequent. This tumor generally affects the peritoneal surface of the pelvis and abdomen; although other less frequent locations have been described testicular location is exceptional. We report the case of a patient presenting with a multicystic mesothelioma of the testicular tunica vaginalis and review the diagnosis, pathology and treatment options for this type of tumor. PMID- 14763425 TI - [Lower massive hematuria deferred by arteriovenous fistula following percutaneous nephrostomy]. AB - OBJECTIVES: To report one case of severe lower hematuria 48 hours after urinary diversion by percutaneous nephrostomy for obstructive uropathy. METHODS: The clinical picture, its severity, and previous procedure probably causing it made the study by arteriography essential to identify the existence of an arteriovenous fistula after stabilization of the acute anaemia; it was confirmed and selective embolization was undertaken with an optimal result. We performed a bibliographic search on Pub-Med (MEDLINE) using the terms "percutaneous nephrostomy", "complications", and "arteriovenous fistula". Most published papers are case reports and series of nephrostomies analyzing their complications and the management of them. RESULTS: Complications attributable to percutaneous nephrostomy are very rare varying from pneumothorax/hydrothorax, infection, perforation and injuries to neighbour organs, allergic reactions to contrasts, urinomas, to pictures of mild to severe hematuria due to vessel laceration and arteriovenous fistula, either high or through bladder. In the case we report as well as in those from the literature, initial treatment of hematuria was watchful waiting; only severity or persistence for more than 3-4 days should make to suspect the existence of arteriovenous fistula, and then arteriography is mandatory to obtain confirmation and if possible treat it by selective embolization; there are various materials to do it. Clinical repercussion was severe but thanks to radiological interventional techniques the case was solved with preservation of the renal unit. CONCLUSIONS: Complications after percutaneous nephrostomy are very rare, and the clinical picture alerts about the possibility of some of them. Radiological tests allow to identify the great majority of them and to perform an effective treatment. PMID- 14763426 TI - [Soft lithiasis ("mous")]. AB - OBJECTIVES: To describe the most relevant features of matrix lithiasis, an infrequent type of lithiasis seldom reported since the generalization of extracorporeal shock wave lithotripsy. METHODS: We report a clinical case and analyze the most important bibliography found by MEDLINE search (1950-1999). We will focus on the physiopathology and etiology of these stones to understand its development and the clinical presentation showed by these patients. RESULTS: Matrix lithiasis is a very rare entity, described almost one century ago. It is mainly composed of lithiasis matrix, and the percentage of mineral content is really low in comparison with usual stones. Persistent urinary tract infection is another constant in these patients, which to some extent conditions the modification of the usual urinary components, creating a series of the disturbances within the urinary tract that favour the aggregation of the components of the lithiasis matrix up to the formation of a stone, without need of aggregation of mineralized crystals. CONCLUSIONS: A high degree of suspicion is necessary for the diagnosis of matrix lithiasis, because it does not show the usual clinical picture of a renal stone. Exhaustive lavage of caliceal cavities and elimination of bacteriuria are essential elements for treatment, in order to avoid recurrence of the disease. PMID- 14763427 TI - A spontaneous giant urinoma: a cause of abdominal mass. AB - OBJECTIVES: To report a case of unilateral ureteral obstruction associated with giant retroperitoneal urinoma which was treated successfully by percutaneous drainage. METHODS: A 19G ultrasound guided needle was introduced into the mass and after dilatation procedure, a 14 Fr nephrostomy catheter was placed and 5.5 of liquid were drained. RESULTS: Drainage from the nephrostomy catheter stopped 24 hours later. Patient was discharged with the nephrostomy tube. One week later, on control tomography, absence of left renal suprahilar posteromedial focal cortical contour was seen. CONCLUSIONS: A minor cortical or forniceal rupture with no urinary extravasation on radiographic examination is thought to be the cause of this urinoma. Percutaneous drainage may be considered as an alternative treatment option to surgical intervention in the treatment of urinomas, especially when there is no persistent ureteral obstruction or communication between the collecting system and the urinoma. PMID- 14763428 TI - [Traumatic emergency. How is our progress?]. PMID- 14763429 TI - [Thin layer preparations in thyroid fine-needle aspiration: study of 200 cases]. AB - INTRODUCTION: Trying to increase the specificity and demonstrated sensibility of thyroid fine needle aspiration (TFNA) in the diagnosis of the thyroid nodule, and to improve the cytomorphologic diagnosis, we used the approach of realization of thin prep (TP), adapting the technique to the methods within our possibilities. MATERIALS AND METHODS: The sample corresponded to a prospective study of 200 cases of material of thyroid nodules, obtained by means of ultrasonography guide fine needle aspiration, over a 30 month period. The study was carried out by means of conventional smear (CS) and TP, cytologic preparations were stained with hematoxilin-eosine (H-E), Papanicolaou (PAP) and Tint 15. The analysis of these same samples was carried out according to sex, age, characteristics of the nodules and cytomorphologic features. RESULTS: The samples were satisfactory in 184 cases (92%) and unsatisfactory in 16 cases (8%). The prevalence in samples corresponded to the female sex (88.5%). The pre-emptive diagnoses corresponded to: Multinodular Goiter: 41 (22.29%)--Hashimoto's thyroiditis: 4 (2.17%)- Follicular lesions I: 86(46.74%), II: 34 (18.47%), III: 6 (3.26%)--Hurthle cell neoplasm: 4 (2.17%)--Papillary carcinoma: 9 (4.90%). The patient study was carried out according to the modified algorithm proposed by Gharib in 1997 and the Department of Cytology of the Karolinska Hospital of Stockholm (Sweden). The cytological diagnosis was compared to the final histological results. CONCLUSIONS: The present work has demonstrated that even with the available methods, we have been able to carry out TP that were acceptable for its evaluation and with the same limitations of those obtained by CS, with respect to preparations dried in direct air and in the cases of non-conclusive diagnoses. We obtained a sensibility of 94.44% and a specificity of 98.79%. PMID- 14763430 TI - Sildenafil citrate for the treatment of patients with cardiovascular diseases with exclusion of coronary artery disease and hypertrophic subaortic stenosis. Its beneficial effect on patients chronic Chagas's and diabetic cardioneuromyopathies, hypertensive and hypertrophic cardiomyopathies, with or without chronic congestive heart failure. AB - Sildenafil citrate is a potent donor of nitric oxide that has been proved to be effective for the treatment of male erectile dysfunction, but it has been contraindicated in patients with cardiovascular diseases, because of sudden death occurred to some of them. Based on the known vasodilator effect of nitrix oxide, effect that should be beneficial for some cardiomyopathies, this work was carried out in order to prove the cardiovascular effects of sildenafil citrate on: 1) heart rate, rhythm and repolarization changes on the ecg; 2) systolic and diastolic arterial blood pressure; 3) left ventricular systolic function and 4) right and left ventricular diastolic function in 26 patients suffering from the following cardiomyopathies: chronic Chagas's and diabetic cardioneuromyopathies, hypertensive and/or hypertrophic cardiomyopathies with or without chronic congestive heart failure. RESULTS: sildenafil citrate 50 mgr after a single oral dose: 1) improved the ecg findings in some patients, worsening the basal ecg in none of the studied patients; 2) significantly reduced systolic and diastolic arterial blood pressure being such reduction very strong in those with basal high level 3) significantly improved left ventricular systolic function in those patients with basal reduced function and 4) Right ventricular diastolic function evaluation: Sildenafil citrate 50 mgr significantly modified to normal pattern the E/A basal altered ratio in those patients with the inverted pattern as well as in those with restrictive pattern of tricupid diastolic influx to the right ventricle during the echo-duplex interrogation (p < 0.0001) 5) Left ventricular diastolic function evaluation: Sildenafil citrate 50 mgr significantly modified to normal pattern the E/A basal altered ratio in those patients with the inverted pattern as well as in those with restrictive pattern of mitral diastolic influx to the left ventricle, during the echo-duplex interrogation (p 0 <.0001). CONCLUSIONS: Based on the above findings it is feasible to propose the use of sildenafil citrate to treat patients with cardiovascular diseases, with exclusion of severe obstructive coronary artery disease, hypertrophic subaortic stenosis and patients with funduscopic alterations that may be affected by a significant and acute increase of flow within the ophthalmic arteries. The right ventricular diastolic changes observed with Sildenafil Citrate may be useful in patients with abnormal right ventricular compliance such as pulmonary stenosis or hypertension. PMID- 14763431 TI - [Semester direct cost by rheumatoid arthritis in patients in a university hospital]. AB - INTRODUCTION: There are no medical publications with economic analysis of rheumatoid arthritis patients (RA) from Argentina are lacking. The objective of the present study is to determine the direct cost and its breakdown in patients with RA. MATERIAL AND METHODS: Fifty-two patients who met the American College of Rheumatology RA criteria were included. Direct cost was calculated over a follow up period of 6 months during year 2001. Variables were analyzed with Student's T test, Mann-Whitney U Test, c' or ANOVA as corresponded. P values < 0.05 were considered significant. RESULTS: The mean monthly home income was $426.6 SD 272. The mean half-yearly direct costs was $677.5 SD 376.2. The components of the direct cost were identified and the mean for medication cost was $606.7 (89%), for lab tests was $45.5 (7%), for medical attention $12.5 (2%) and other costs $2.4. No differences in total cost or in medication cost were found when compared considering age, evolution time of RA or HAQ scores. CONCLUSION: Half-yearly direct cost in RA is excessively high considering the monthly mean income of the patients being analyzed. The cost of medication was the principal component of the direct cost. PMID- 14763432 TI - [Prevalence of pediculus humanus capitis infestation in schoolchildren at Despenaderos, Cordoba Province]. AB - BACKGROUND: Approximately 80% of Argentine children have pediculosis, making it imperious to detect the defects in the prevention and control actions for this parasitosis. OBJECTIVES: To study the infestation level by P. h. capitis, its temporal variation, and the association between infestation and host characteristics. MATERIALS AND METHODS: A monthly census in children of 6-8 years old from 2 primary schools (state and private) was carried out, recording the parasite stages found, children characteristics (sex, colour and length of hair), and infestation level. Data were analysed by means of contingency tables, difference of proportions, quotient of chances, and rank correlation coefficient. The independence of variables was analysed by c' test. RESULTS: The prevalence and chances of becoming infested were higher at the state school, where also all infestation levels were recorded, which did not occur at the private institute. The frequency of children infested was significantly lower when they had short hair. The correlation among infestation degree and hair length (short-long) was only significant for boys. Pediculosis cases were recorded at the beginning of the school year in both educative institutions, which demonstrated that infestation was acquired not only at school but also at home. CONCLUSIONS: Pediculosis is a social problems and the whole community must participate in preventive and control actions, for which sanitary education is essential, and the myth that the school is the only place of contagion must be eradicated. PMID- 14763433 TI - [Infection outbreak due to resistant Enterococcus faecium to glycopeptides in a Hospital of Rosario, Argentina]. AB - Enterococci causes serious illness in immunocompromised patients and in severely ill, hospitalized patients. They are commonly isolated from the human gastrointestinal tract, and resistance to vancomycin has increased in frequency during the past few years. We report three cases of nosocomial vancomycin resistant Entrococcus faecium (VRE) infections from September to December of 2000. A 54-year-old man presented post surgical wound infection due to VRE after 2 months of hospitalization. The second is a 65-year-old neutropenic patient with a medical history of acute myeloid leukemia and esophageal carcinoma. In this case, central venous catheter was colonized with EVR. The third, a 65-year-old woman who received therapy with ceftriaxone, ciprofloxacin and clindamicin had a urinary tract infection due to EVR. The three isolates were resistant to vancomycin (MIC value > 256 micrograms/ml). The DNA amplification pattern obtained by OD-PCR were similar in the three cases. PMID- 14763434 TI - [Present state in the surgical treatment of acute cecal diverticulitis]. AB - BACKGROUND: Cecal acute diverticulitis is a rare disease with symptoms similar to acute appendicitis and is therefore difficult to diagnose. OBJECTIVES: To evaluate therapeutical conduct according to clinical for ms of presentation and the intraoperative findings. LOCATION: Emergency Service of Clinical National Hospital. U.N.C. (Cordoba National University), and the ABC group. DESIGN: Retrospective observation. MATERIALS AND METHODS: Twenty-four patients with cecal diverticulitis were treated surgically, of which seventeen were men and seven were women with an average age of 47 years. The first group was formed with 16 patients and the second with 8. All patients presented abdominal pain, fever, nausea and vomiting when they were admitted, but one showed enterorrhagia and 2 abdominal distention and ileus. Physical examination produced pain and tenderness in RLQ, but in 6 peritoneum was not involved and in 4 a tumor was palpable in that region. Two patients, had a perforating acute abdominal condition. RESULTS: Sixteen right hemicolectomies were performed. In the remaining eight a cecostomy was decided. One patient died due to TEPA (Acute pulmonary thromboembolism). Abdominal wall abscesses appeared in six patients, and in four of them an incisional middle hernia appeared in postoperative period. CONCLUSIONS: We consider that in those patients presenting serious problems in deciding whether the inflammatory condition was benign or malignant, or when perforated cecal diverticulum with vascular compromise and subsequent cecal wall necrosis has been suspected, we believe that the resection is the best treatment. PMID- 14763435 TI - [Acute carotid thrombosis in penetrating traumatism: is revascularization necessary in the patient without central neurological deficit?]. AB - INTRODUCTION: Carotid lesions require priority in both evaluation and treatment due to their high morbidity and mortality. Controversy about therapeutic behavior in these patients with or without central neurological deficit is still under in discussion. OBJECTIVES: To present a patient with acute carotid thrombosis due to a shotgun wound and discuss its therapeutic behavior. SETTING: Hospital de Urgencias in Cordoba city. MATERIAL AND METHODS: A 15-year-old male patient is presented with a "point-blank" shotgun wound in the soft parts of the left cervical region, and a left carotid thrombosis with no central neurological deficit. RESULTS: Wound toilette and carotid revascularization by means of resection and venous by-pass with external carotid ligature was performed. The procedure was finished by delaging for plastic reconstruction of the cervical injury. Carotid postoperative angiographic control showed good permeability with no carotid flow alteration. CONCLUSION: Penetrating carotid injuries should be resolved, if technically possible, with revascularization of the carotid sector. This procedure has to be aborted if the patient is in coma or the lesion is difficult to repair, in such a case ligature should be carried out. PMID- 14763436 TI - [Pelvic inflammatory disease associated with Neisseria meningitis bacteremia]. AB - The Pelvic Inflammatory disease (PID) caused by Neisseria meningitidis (Nm), as an extraneurological manifestation considered as a case study with few precedents. It is so far known that the occasional transposition of ecological niches occurs. There are several bibliographical reports that show an increase in the colonization of the genitourinary tract due to Nm. These may or may not cause disease in individuals during colonization. In adults, the symptoms and any complications which may arise are similar to those caused by Neisseria gonorrhoeae (Ng). It is highly necessary to correctly identify the agents of Neisseria gender which are obtained from the genital tract. This permits an accurate diagnosis to be made and results in a better understanding of the epidemiology. Morbimortality arising from Nm infection is worth studying to produce a better knowledge of the pathogenia of the infection caused by this microorganism. PMID- 14763438 TI - [Thyroid diseases]. PMID- 14763437 TI - Mitotracker green is a P-glycoprotein substrate. AB - P-glycoprotein has a widespread expression on normal tissues. The protein has also been strongly associated with the multidrug resistance phenotype (MDR) on tumor cells. The employment of flow cytometry and confocal microscopy has contributed to the discovery and application of new particular fluorescent dyes. Nevertheless, several studies are being performed in different cellular types neglecting the expression/activity of MDR proteins. Because many fluorochromes have been reported as P-glycoprotein substrates, an especial attention must be given to the properties of new dyes in the presence of MDR proteins. Flow cytometric analyzes of Mitotracker Green (MTG) fluorescence profile were performed in a human erythroleukemic cell line and its resistant counterpart. In this report we demonstrated that MTG, a probe used to evaluate the mitochondrial mass, is a P-glycoprotein substrate and its staining profile is dependent on the activity of this protein. In vitro studies on a human erythroleukemic cell line and its resistant counterpart revealed that MDR modulators (Cyclosporin A, Verapamil, and Trifluoperazine) alter the MTG fluorescence pattern on a resistant cell line. The findings suggest that attention should be given to the expression of P-glycoprotein when performing an evaluation of mitochondria properties with MTG. PMID- 14763443 TI - "Spreading sanitary enlightenment": Race, identity, and the emergence of a creole medical profession in British Guiana. PMID- 14763444 TI - An insidious nosocomial outbreak due to Salmonella enteritidis. AB - OBJECTIVES: To investigate an increase in the number of Salmonella enteritidis isolates detected in a large hospital to ascertain whether it was due to a nosocomial source, to identify the mechanisms of transmission, and to institute effective control measures to prevent future episodes. DESIGN: Observational study, survey of all microbiological samples positive for S. enteritidis detected in the hospital, outbreak investigation, and review of the literature. SETTING: A tertiary-care teaching hospital for adults in Barcelona, Spain. RESULTS: During a 7-month period from May to November 1998, we identified 22 inpatients with S. enteritidis infection for whom nosocomial acquisition was strongly suspected. The attack rate was 0.138 per 1,000 patient-days. All affected patients were immunosuppressed and overall mortality was 41% (9 of 22). A sample of a meal cooked in the kitchen was culture positive for S. enteritidis. All isolates shared the same antibiotic susceptibility pattern and all except one shared the same pulsed-field gel electrophoresis (PFGE) pattern, but PFGE could not differentiate between outbreak-related and control strains. After compliance with kitchen hygiene procedures was emphasized and cleansing was intensified, no more cases were detected. CONCLUSIONS: Apparently, sporadic cases of S. enteritidis may be part of an outbreak with a low attack rate. A small but persistent inoculum affecting only individuals with special predisposition for Salmonella infection might account for this. Suspicion should be raised in hospitals and institutions with a highly susceptible population. PMID- 14763445 TI - [Renal toxicity of Aristolochia and its prevention]. PMID- 14763446 TI - [Advances in the study of Ilex pubescens Hook. et Arm: chemical constituents and pharmacological action]. PMID- 14763447 TI - Matrix metalloprotenase-9 in acute pyelonephritis. PMID- 14763449 TI - Bibliography of toxinology. PMID- 14763448 TI - Building user involvement in motor neurone disease: key lessons. PMID- 14763450 TI - Endocrine-related resources from the National Institutes of Health. PMID- 14763452 TI - Faculty matters. Karen H. Morin. PMID- 14763453 TI - What do Internet-based alcohol treatment websites offer? AB - The Internet was searched for websites that advertised or provided treatment or help for alcohol problems. Websites were evaluated for the types of treatment offered and whether the treatment had an empirical basis. While a wide range of treatments were advertised, very few websites offered online services. In addition, very few sites provided or advertised alcohol treatment programs that were empirically based. Recommendations for future Internet-based health care and treatment are offered. PMID- 14763454 TI - [Standards of visceral organ procurement]. AB - Due to poor willingness in the population to donate organs and increasing number of patients on the waiting list for organ transplantation there is an imminent need on cadaveric organs. Although efforts have been to optimize the prerequisites for organ donation, less than 50% of possible organ harvesting procedures take place. In our country, only 13.1 organ donations .per 1 million inhabitants were realized in 2001. These data force the medical community to maximize the utility of the donor pool. A refined surgical technique, increased efficacy of solutions for graft conservation, an advanced perioperative management and new potent immunosuppressive drugs contributed to a post transplant 1-year graft and patient survival of up to 80 to 90%. The quality of a graft has a significant impact on the outcome of transplantation. An optimal graft function will not only be achieved with an adequate surgical and preservation technique but also with an appropriate management of the brain-dead donor on the intensive care unit. Basic knowledge of brain death definition, insight into the organization of organ donation, management of the donor, and modern surgical harvesting techniques are presented. PMID- 14763455 TI - [Hans Erhard Bock (on his 100th birthday on 12/31/2003)]. PMID- 14763456 TI - [H. E. Bock and the German Society of Internal Medicine]. PMID- 14763457 TI - Nurses advocate for nurses. PMID- 14763458 TI - [Evolution of pharmaceutical regulations from the origins to Germinal law]. AB - The rising of pharmaceutical regulations took place in the civilisations located around the Mediterranean Sea. Egypt, Mesopotamia, Greco-Roman world, Byzantine Empire, were followed by Bagdad, where sayadila, inspections and grabadins appeared. The creation of Universities, during the XIIth and XIIIth centuries, plaid a role in the rising of apothecaries communities in Occident. Melfi Constitutions (Constititiones Melfiae, 1231) and new constitutions (1241) inspired most of the regulations of apothecaries communities in Europe. Declaration of the King of France in April 1777 announced the famous "Loi de Germinal", Germinal law, which organized the modern Pharmacy, in 1803. PMID- 14763459 TI - [Psychiatry and thoughts of a novelist]. PMID- 14763460 TI - Cardiac troponin in dialysis patients. PMID- 14763461 TI - EAC training and medical support for International Space Station astronauts. AB - The operation of the International Space Station (ISS) will be a global multilateral endeavour. Each International Partner will be responsible for the operation of its elements and for providing a crew complement proportional to its share of the overall resources. The preparations of the European Astronaut Centre to furnish training and medical support for the ISS astronauts are described. PMID- 14763462 TI - Ten years of the European Astronaut Centre (EAC). AB - The European Astronaut Centre, the home base of ESA's Astronaut Corps, celebrated its 10th anniversary on 17 May 2000 with a media event highlighting the past, present and future of the Agency's manned space programme. PMID- 14763463 TI - Clinical study on the metastasis to the eyes from breast cancer. AB - In our hospital, 24 patients who underwent surgery for breast cancer during 1980 to 2001 were diagnosed with metastasis to the eye. Metastasis to the choroid was found most frequently, making the choroid the most common site of metastasis. A few patients had metastasis to the orbit. Decreased visual acuity and tunnel vision were frequently found in patients who had metastasis to the choroid, and ocular floaters and blurred vision were also found in a few patients. Patients with metastasis to the orbit showed diplopia caused by ocular dyskinesia and eyelid swelling. The mean postoperative period until the diagnosis with metastasis to the eye was 3 years and 2 months, with most cases diagnosed between 20 and 40 months postoperatively, a relatively long period. We performed radiotherapy in 21 of the 24 patients, and more than half of the patients showed improvement. The mean survival period after diagnosis with metastasis to the eye was 10 months, and some of them already had recurrence to other organs such as the bones or lungs. Examination with consideration of metastasis to the eyes is required to improve the quality of life of cancer patients,. PMID- 14763464 TI - Opisthorchis viverrini and opisthorchiasis: The 21st Century review. Proceedings of the Congress of Opisthorchiasis and Cholangiocarcinoma. May 28-30, 2002. Khon Kaen, Thailand. PMID- 14763465 TI - Development of a questionnaire to assess subjective vision score in myopes seeking refractive surgery. AB - PURPOSE: To assess vision in patients with myopia and myopic astigmatism before and after refractive surgery. METHODS: A prospective controlled study of visual quality amongst myopes and astigmatic myopes. Focus groups, ophthalmic surgeons, and questionnaire experts devised a Subjective Vision Questionnaire (SVQ), modified after a pilot trial. Participants were administered the SVQ before clinical evaluation. Items answered by over 95%, with factor loadings > 0.55 were included. Test-retest reliability was assessed by repeat testing. Factor analysis identified groups of questions measuring particular dimensions of data. RESULTS: Sixty-seven items were answered by 128 patients and reduced to 24 items in a final questionnaire. Factor analysis identified six types of questions within the questionnaire, the most important of which was related to driving. CONCLUSION: The simplicity, low cost, and psychometric properties of the Subjective Vision Questionnaire support its use clinically and in research. PMID- 14763466 TI - Refractive lens exchange versus iris-claw Artisan phakic intraocular lens for hyperopia. AB - PURPOSE: To study a paired-match comparison between refractive lens exchange with pseudophakic IOL implant (RLE) and Artisan phakic IOL for high hyperopia. METHODS: Nineteen eyes (12 patients, 20 to 41 years old) with an Artisan phakic IOL (Model 203: 1.00-D increment) for hyperopia from +2.75 to +9.25 D were matched to 19 eyes (15 patients, 26 to 46 yr) with hyperopia from +2.75 to +7.50 D, who had refractive lens exchange (pseudophakic IOL implantation; lenses: 0.50 D increment). Average paired-match difference was 1.13 D and 7.7 years of age. RESULTS: At 1 month after surgery, 84% of refractive lens exchange/pseudophakic IOL eyes and 94% of Artisan phakic IOL eyes had a spherical equivalent refraction within +/- 1.00 D of emmetropia; 58% and 68% of eyes, respectively, were within +/- 0.50 D (P = .97). No eye lost lines of best spectacle-corrected visual acuity (BSCVA) and no significant changes in BSCVA were found in any eye at 1 month after surgery (P = .17). The percentage of eyes with uncorrected visual acuity (UCVA) of 20/40 or better improved from 79% to 89% of eyes at 1 to 2 months after phakic IOL; it remained at 89% to 82% of eyes from 1 to 2 months after refractive lens exchange/ pseudophakic IOL. The coefficient of correlation showed statistically better accuracy (intended vs. achieved refraction; P = .035) for the Artisan phakic IOL (R = 0.83) than for refractive lens exchange/ pseudophakic IOL (R = 0.50). CONCLUSIONS: Spherical equivalent refraction outcome and BSCVA after surgery were similar for both procedures. The Artisan phakic IOL in carefully selected patients provided a better overall outcome for young patients with high hyperopia whose accommodation was preserved, as compared to refractive lens exchange. PMID- 14763467 TI - Laser in situ keratomileusis improves visual acuity in some adult eyes with amblyopia. AB - PURPOSE: To report the results of laser in situ keratomileusis (LASIK) in a series of adult patients with amblyopia. METHODS: A retrospective noncomparative review was performed on patients with amblyopia who underwent LASIK for correction of ametropia, using the Summit Krumeich Barraquer microkeratome and the Nidek EC-5000 excimer laser. Data were collected on uncorrected visual acuity, best spectacle-corrected visual acuity, manifest refraction, anterior segment evaluation, intraocular pressure, corneal topography, and dilated fundus examination (preoperative and postoperatively on day 1, months 2 and 6). RESULTS: Eight eyes of seven patients were included, with a mean patient age of 30 +/- 10 years (range 21 to 49 yr). Mean preoperative spherical equivalent refraction was 4.70 D (range -12.62 to +4.71 D) and the best spectacle-corrected visual acuity varied from 20/32 to 20/80. At 2 months after LASIK, mean spherical equivalent refraction was -0.37 +/- 0.60 D (range -1.37 to + 0.60 D), uncorrected visual acuity ranged between 20/20 to 20/30, and a mean gain of 3 Snellen lines (range 2 to 4 lines) was observed. All patients reported significant subjective improvement in their perception of vision. The visual acuity and subjective improvement were maintained throughout 6 months postoperatively. CONCLUSIONS: LASIK with the Nidek EC-5000 laser for correction of ametropia in adult amblyopic eyes provided encouraging results for visual acuity improvement beyond correction of the refractive error. PMID- 14763468 TI - Comparison of four corneal pachymetry technique;s in corneal refractive surgery. AB - PURPOSE: To assess the effectiveness of optical and ultrasonic pachymetry when examining corneas of patients before and after laser in situ keratomileusis (LASIK). METHODS: We conducted a prospective study of 25 patients (50 eyes) who had LASIK for myopia or myopic astigmatism (mean spherical equivalent refraction 4.80 +/- 3.60 D; range -1.75 to -12.00 D). Corneal thickness was measured using 1) conventional ultrasonic pachymetry (20 MHz probe) in the central cornea, 2) ASL confocal microscopy (CMTF analysis), 3) Orbscan II Analyzer, and 4) ultrasound biomicroscopy (UBM) (50 MHz probe) preoperatively and 3 months postoperatively. RESULTS: Preoperative pachymetry values measured with the Orbscan II Analyzer (551.3 +/- 27.3 microm) were almost identical to those measured with the 20 MHz ultrasonic probe (553.7 +/- 25.7 microm; t-test P = .652). The mean preoperative pachymetry for UBM was 566.7 +/- 38.8 microm and for CMTF was 553.2 +/- 15.5 microm. The optic and ultrasonic system measurements differed in operated eyes; 20 MHz ultrasonic pachymetry yielded an average 478.7 +/- 23.5 microm and the UBM, 482.4 +/- 40.3 microm. These measurements differed from the Orbscan II (Tukey test; P = .023) measurement of 461.1 +/- 37.5 microm but not from the confocal microscopy measurement of 475.2 +/- 41.8 microm. UBM registered greater values that were more variable than those obtained with ultrasonic pachymetry. CONCLUSIONS: In non-operated eyes, corneal thickness was similar using conventional ultrasound, Orbscan II, or confocal techniques, with ultrasound biomicroscopy giving thicker readings. PMID- 14763469 TI - Wavefront-guided laser in situ keratomileusis with the Bausch & Lomb Zyoptix system. AB - PURPOSE: To evaluate the clinical results of wavefront-guided laser in situ keratomileusis (LASIK) with the Zyoptix system. METHODS: Twelve patients (24 eyes) underwent wavefront-guided LASIK with the Bausch & Lomb Zyoptix system. Uncorrected and best spectacle-corrected visual acuity and manifest refraction were measured at postoperative day 1, week 1, and months 1 and 3. A subjective vision quality questionnaire evaluated light sensitivity, dryness, tearing, glare, halos, ghost images, and difficulties in night driving, preoperatively and 3 months postoperatively. RESULTS: Preoperatively, mean sphere was -3.70 +/- 2.33 D (range -0.50 to -8.00 D), mean cylinder was -0.90 +/- 0.98 D (range 0 to -3.00 D), and mean spherical equivalent refraction was -4.15 +/- 2.16 D (range -1.38 to -8.25 D). Three-month postoperative spherical equivalent refraction was within +/ 0.50 D of emmetropia in 17 eyes (70.8%) and within +/- 1.00 D in 22 eyes (91.7%). At 3 months postoperatively, no eyes lost any lines of BSCVA and eight eyes (33.3%) gained 2 lines. The ratio of postoperative BSCVA to preoperative BSCVA (safety) was 1.05 +/- 0.09 (range 1.00 to 1.20) at 1 month and 1.07 +/- 0.10 (range 1.00 to 1.29) at 3 months. The ratio of postoperative UCVA to preoperative BSCVA (efficacy) was 0.96 +/- 0.12 (range 0.80 to 1.20) at 1 month and 0.95 +/- 0.12 (range 0.8 to 1.2) at 3 months. The subjective vision quality questionnaire revealed less tearing, fewer halos, and less difficulty in night driving after wavefront-guided LASIK. Comparison of higher order optical aberrations before and after surgery was not done. CONCLUSIONS: Wavefront-guided LASIK with the Bausch & Lomb Zyoptix system was safe and effective in correcting low to moderate myopic refractive error. PMID- 14763471 TI - Retreatment to enlarge small excimer laser optical zones using combined myopic and hyperopic ablations. AB - PURPOSE: We describe a retreatment technique using a combination of large diameter myopic and hyperopic excimer laser ablations of a near equivalent diopter value to enlarge previous small ablation zones without altering the refractive result obtained by the initial surgery. METHODS: Eight eyes of six patients were retreated with a Technolas Keracor 217 laser in order to enlarge the optical zone. All patients had a good refractive result after the initial surgery, but reported halos under low light conditions. Retreatment consisted of a combination of myopic and hyperopic ablations of near equivalent value, eg, 1.00 D and +1.00 D treatment. These opposite value ablations should neutralize their respective refractive effect, but by removing a larger diameter of tissue, actually displace the transition zones further toward the corneal periphery. Although the initial treatment used photorefractive keratectomy in four of the eight eyes, LASIK was used for retreatment in all eyes. Improvement was assessed subjectively by the patient and was measured objectively with a Technomed C-Scan ray-tracing program. RESULTS: In six of eight eyes, patients experienced moderate or marked improvement. This improvement was objectively correlated on postoperative corneal topography. CONCLUSION: Combined larger diameter myopic and hyperopic retreatment seems to be safe and effective for enlarging previous small treatment zones, reducing symptoms such as halos in selected patients. PMID- 14763470 TI - Effect of suturing on latrogenic keratectasia after laser in situ keratomileusis. AB - PURPOSE: To evaluate the influence of corneal suturing on the clinical course of iatrogenic keratectasia after laser in situ keratomileusis (LASIK), in order to improve quality of vision and to avoid progressive deterioration of visual acuity. METHODS: Three patients (four eyes) who had a perforation during LASIK or became keratectatic after LASIK received corneal suturing with 10-0 nylon immediately or up to 11 days after LASIK. Sutures were left in place for 1 to 4 months. Patients were followed for 20 to 23 months after suturing. RESULTS: All eyes had a significant improvement in uncorrected and best spectacle-corrected visual acuity after suturing. The improvements in visual acuity for all patients remained unchanged after suture removal. In patient 1, keratectatic changes were not detected in the sutured eye although they were detected in the contralateral unsutured eye. In patient 2, both eyes were sutured, one immediately and the other 11 days after LASIK; no keratectatic changes were noted in either eye. Slight progression of corneal protrusion was observed in patient 3, who had corneal suturing after development of prominent keratectasia 2 weeks after LASIK. This eye showed stable but moderate keratectatic change 9 months after suture removal. CONCLUSIONS: Flap suturing in patients during the course of keratectasia after LASIK improved both aided and unaided visual acuity. Prompt suturing seemed to provide a means of preventive management for iatrogenic keratectasia in anticipation of keratoplasty. PMID- 14763472 TI - Diode laser thermal keratoplasty to correct hyperopia. AB - PURPOSE: We investigated long-term efficacy, predictability, stability, and safety of diode laser thermal keratoplasty (DTK) to correct hyperopia. METHODS: DTK was performed on 24 eyes (18 patients). Eight eyes with high hyperopia (mean +4.75 +/- 0.63 D; range +3.50 to +5.50 D) received 12 pairs of coagulation spots at 6-mm and 7-mm treatment zone diameters; eight eyes with low hyperopia (mean +2.25 +/- 0.40 D; range +1.50 to +2.75 D) received eight coagulation spots at 8 mm, and eight eyes with low hyperopia (mean +1.50 +/- 0.46 D; range +1.25 to +2.25 D) were treated to induce mild myopia (-1.50 D) in the non-dominant eye for monovision using eight pairs of spots at 7 and 8-mm diameters. Minimum follow-up was 18 months. RESULTS: Mean decrease in cycloplegic refraction at 18 months was 5.00 +/- 0.38 D in the high hyperopia group, 1.75 +/- 0.19 D in the low hyperopia group, and 3.25 +/- 0.27 D in the presbyopia group. Mean increase in uncorrected visual acuity (UCVA) at 18 months was 8.125 +/- 2.1 Snellen lines in the high hyperopia group, 6.625 +/- 0.744 lines for low hyperopia; decrease of 1.00 +/- 1.85 line occurred in the presbyopia group. Near UCVA in the presbyopia group improved by 3.875 +/- 0.83 Jaeger lines. Best spectacle-corrected visual acuity (BSCVA) was restored by 3 months in all eyes. CONCLUSION: DTK was an effective and fairly safe procedure, with reasonable predictability and stability. Nomograms for laser energy level, treatment zone diameter, and number of spots need improvement. PMID- 14763473 TI - Management of the ocular surface and tear film before, during, and after laser in situ keratomileusis. AB - PURPOSE: To identify evidence-based, best practice strategies for managing the ocular surface and tear film before, during, and after laser in situ keratomileusis (LASIK). METHODS: After a comprehensive review of relevant published literature, evidence-based recommendations for best practice management strategies are presented. RESULTS: Symptoms of ocular irritation and signs of dysfunction of the integrated lacrimal gland/ocular surface functional gland unit are common before and after LASIK. The status of the ocular surface and tear film before LASIK can impact surgical outcomes in terms of potential complications during and after surgery, refractive outcome, optical quality, patient satisfaction, and the severity and duration of dry eye after LASIK. Before LASIK, the health of the ocular surface should be optimized and patients selected appropriately. Dry eye before surgery and female gender are risk factors for developing chronic dry eye after LASIK. Management of the ocular surface during LASIK can minimize ocular surface damage and the risk of adverse outcomes. Long term management of the tear film and ocular surface after LASIK can reduce the severity and duration of dry eye symptoms and signs. CONCLUSIONS: Strategies to manage the integrated ocular surface/lacrimal gland functional unit before, during, and after LASIK can optimize outcomes. As problems with the ocular surface and tear film are relatively common, attention should focus on the use and improvement of evidence-based management strategies. PMID- 14763474 TI - Risk factors for development of diffuse lamellar keratitis after laser in situ keratomileusis. AB - PURPOSE: To investigate the risk factors and mechanisms of diffuse lamellar keratitis (DLK) after laser in situ keratomileusis (LASIK). METHODS: In 5708 eyes of 2927 patients who underwent LASIK, the patient's background, clinical findings, and surgical data were retrospectively evaluated and compared with patients who had DLK and those who did not (non-DLK group). RESULTS: DLK occurred in 46 of 5708 eyes (0.81%). DLK occurred more frequently in eyes operated with the MK-2000 microkeratome (1.1%) than with the LSK-One microkeratome (0.6%; P = .039). Corneal endothelial cell density (2686.8 +/- 235.3/mm2) was significantly lower in eyes that developed DLK than in eyes without DLK (2783.8 +/- 275.5/mm2; P = .017). The width of palpebral fissure in a normal state and the maximum opening position were significantly larger in the DLK group (10.3 +/- 1.9 mm and 18.1 +/- 7.2 mm, respectively) than the non-DLK group (8.3 +/- 1.6 mm with P < .001; 11.5 +/- 2.0 mm with P = .012). CONCLUSIONS: The type of microkeratome, lower corneal endothelial cell density, and larger palpebral fissure were potential risk factors for DLK after LASIK. These factors may be associated with delayed wound healing of the corneal flap margin, resulting in poor sealing of the flap, which may induce the influx of inflammatory cells. PMID- 14763475 TI - Prospective, randomized trial of diclofenac and ketorolac after refractive surgery. AB - PURPOSE: To compare the effectiveness of diclofenac and ketorolac in relieving corneal pain after refractive surgery, and determine if there is a difference in stinging on instillation. METHODS: Thirty patients were randomized prospectively to postoperative diclofenac in one eye and ketorolac in the other. Patients and surgeon did not know which medications were used. Ocular postoperative pain and discomfort on instillation of medication were measured after radial keratotomy with a visual analog scale and a questionnaire. RESULTS: Both medications were highly effective in relieving pain. There was no significant difference in pain relief, or stinging on instillation (P = .29). CONCLUSION: There was no statistical difference in the effectiveness of the medications on pain relief, or in stinging on instillation. PMID- 14763476 TI - Laser intrastromal keratoplasty--case report. AB - PURPOSE: To evaluate the feasibility of correcting high hyperopia by means of intrastromal implantation of a laser shaped corneal lenticule prepared from a human donor eye. METHODS: A female patient with high hyperopia and irregular astigmatism resulting from multiple laser in situ keratomileusis procedures and lamellar keratoplasty underwent laser intrastromal keratoplasty. Her preoperative uncorrected visual acuity (UCVA) was 20/300 and best spectacle-corrected visual acuity (BSCVA) was 20/100 with a refraction of +8.00 -1.00 x 130 degrees. Corneal topography showed a highly irregular corneal surface. Central corneal thickness was 398 microm. Lenticule preparation included mechanical de-epithelialization of a human donor eye, keratectomy with a microkeratome, user-designed software combining a photorefractive keratectomy (PRK) treatment for +8.00 D sphere, an ablation zone of 7.0 mm, and a circumferential cut (internal diameter of 6.5 mm) for tissue ablation. Implantation involved re-lifting the flap, positioning the lenticule onto the corneal bed, and repositioning of the flap. RESULTS: The operation was uneventful as was the early postoperative follow-up. BSCVA improved to 20/50 with +1.00 -2.25 x 120 degrees at 2 months postoperatively. Corneal topography showed a more regular cornea with increased curvature in all meridians. Central corneal thickness increased to 600 microm. CONCLUSION: Laser intrastroma keratoplasty may be an option for correcting high hyperopa and irregular astigmatism in eyes with a thin corneal bed. PMID- 14763477 TI - Macular hole after LASIK. PMID- 14763478 TI - More to the mysterious tale: the search for the cause of 100+ cases of diffuse lamellar keratitis. PMID- 14763479 TI - Improved alcohol solution for LASEK. PMID- 14763480 TI - Pupil dilation and wavefront aberration. PMID- 14763481 TI - Microkeratome-induced reduction of astigmatism after RK. PMID- 14763482 TI - Laser epithelial keratomileusis (LASEK) for myopia in patients with a thin cornea. PMID- 14763483 TI - Late onset decreased vision in a steroid responder after LASIK associated with interface fluid. PMID- 14763484 TI - Artificial lung: bench toward bedside. PMID- 14763485 TI - The artificial lung/inflammatory interface: a refocus of the problem. PMID- 14763486 TI - Effects of basic fibroblast growth factor and transforming growth factor-beta on maturation of human pediatric aortic cell culture for tissue engineering of cardiovascular structures. AB - Optimal in vitro conditions are necessary for the development of a strong, well structured, and functional tissue engineered cardiovascular structure eventually designed for implantation. To further optimize in vitro conditions for cell proliferation and extracellular matrix formation in tissue engineering of cardiovascular structures, in this study, ascorbic acid and growth factors as additives to standard cell culture medium were evaluated for their effect on tissue development in vitro. Biodegradable polymer patches [polyglycolic acid (PGA) coated with poly-4-hydroxybutyrate (P4HB)] were seeded with human pediatric aortic cells and cultured for 7 and 28 days. Group A was cultured with standard medium (DMEM with 10% fetal calf serum and 1% antibiotics) supplemented with ascorbic acid; group B was cultured with standard medium plus ascorbic acid and basic fibroblast growth factor (bFGF); group C was cultured with standard medium adding ascorbic acid and transforming growth factor (TGF). Analysis of the cell seeded polymer constructs included DNA assay, collagen assay, and histologic and immunohistochemical examination for cell proliferation and collagen formation. After 7 and 28 days of culture, group B and group C showed a significantly higher DNA content compared with group A. The addition of bFGF (group B) led to a markedly higher collagen synthesis after 28 days of culture compared with the additives in groups C and A. The histologic and immunohistochemical examination also revealed a more dense, organized tissue development with pronounced matrix protein formation in the tissue engineered structures in group B after 28 days of culture. When seeded on to the polymeric scaffold, human vascular cells proliferate and form organized cell tissue after 28 days of culture. The addition of bFGF and ascorbic acid to the standard medium enhances cell proliferation and collagen synthesis on the biodegradable polymer, which leads to the formation of more mature, well organized tissue engineered structures. PMID- 14763487 TI - Hemodynamic and pressure-volume responses to continuous and pulsatile ventricular assist in an adult mock circulation. AB - This study investigated the hemodynamic and left ventricular (LV) pressure-volume loop responses to continuous versus pulsatile assist techniques at 50% and 100% bypass flow rates during simulated ventricular pathophysiologic states (normal, failing, recovery) with Starling response behavior in an adult mock circulation. The rationale for this approach was the desire to conduct a preliminary investigation in a well controlled environment that cannot be as easily produced in an animal model or clinical setting. Continuous and pulsatile flow ventricular assist devices (VADs) were connected to ventricular apical and aortic root return cannulae. The mock circulation was instrumented with a pressure-volume conductance catheter for simultaneous measurement of aortic root pressure and LV pressure and volume; a left atrial pressure catheter; a distal aortic pressure catheter; and aortic root, aortic distal, VAD output, and coronary flow probes. Filling pressures (mean left atrial and LV end diastolic) were reduced with each assist technique; continuous assist reduced filling pressures by 50% more than pulsatile. This reduction, however, was at the expense of a higher mean distal aortic pressure and lower diastolic to systolic coronary artery flow ratio. At full bypass flow (100%) for both assist devices, there was a pronounced effect on hemodynamic parameters, whereas the lesser bypass flow (50%) had only a slight influence. Hemodynamic responses to continuous and pulsatile assist during simulated heart failure differed from normal and recovery states. These findings suggest the potential for differences in endocardial perfusion between assist techniques that may warrant further investigation in an in vivo model, the need for controlling the amount of bypass flow, and the importance in considering the choice of in vivo model. PMID- 14763488 TI - Collected nondimensional performance of rotary dynamic blood pumps. AB - "Nonpulsatile" or "continuous flow" blood pumps are a relatively new application of the rotary dynamic blood pumping principle. They fall outside the normal envelop of pumps, considering their small size, viscosity of the fluid pumped, need for particularly good internal flow patterns, and desire for high efficiency. This article establishes the state of the art in the field of blood pump performance. Trends in efficiency, shut off pressure coefficient, and nondimensional power behavior as a function of nondimensional flow are identified. Blood pumps show agreement with the published effects of low Reynolds numbers in conventional pumps. PMID- 14763489 TI - Is center specific implantation volume a predictor of clinical outcomes with mechanical circulatory support? AB - This study examined the relationship between implant center volumes and survival rate for implantation of mechanical circulatory support (MCS) devices. Using the Novacor left ventricular assist system (LVAS) Registry, the study cohort consisted of 1,348 patients with established outcomes from 97 centers stratified by the implant center volume: 1-10 implants (n = 199, 65 centers), 11-25 implants (n = 189, 12 centers), 26-50 implants (n = 445, 13 centers), and more than 51 implants (n = 515, 7 centers). Regression and correlation analyses were performed. Regression analysis found a negative impact on survival for centers performing 1-10 implants, with an odds ratio of 1.73 (95% confidence interval, 1.28-2.34; p < 0.001). Composite results from the first 10 implants of each larger volume center were then compared with the group with 1-10 implants, demonstrating that centers with larger volumes had superior results, even in the early patient experience (61% versus 46% transplanted/weaned, p < 0.001). However, when annualized outcomes (i.e., outcomes by calendar year) were determined for each center, no significant correlation was found between the outcomes and annualized frequency of implantation (R2 = 0.003, n = 422). Although the total number of implants performed at a specific center appeared to impact clinical outcomes, no correlation was found between annualized frequency of implant and clinical outcome. PMID- 14763490 TI - Characterization of an adult mock circulation for testing cardiac support devices. AB - A need exists for a mock circulation that behaves in a physiologic manner for testing cardiac devices in normal and pathologic states. To address this need, an integrated mock cardiovascular system consisting of an atrium, ventricle, and systemic and coronary vasculature was developed specifically for testing ventricular assist devices (VADs). This test configuration enables atrial or ventricular apex inflow and aortic outflow cannulation connections. The objective of this study was to assess the ability of the mock ventricle to mimic the Frank Starling response of normal, heart failure, and cardiac recovery conditions. The pressure-volume relationship of the mock ventricle was evaluated by varying ventricular volume over a wide range via atrial (preload) and aortic (afterload) occlusions. The input impedance of the mock vasculature was calculated using aortic pressure and flow measurements and also was used to estimate resistance, compliance, and inertial mechanical properties of the circulatory system. Results demonstrated that the mock ventricle pressure-volume loops and the end diastolic and end systolic pressure-volume relationships are representative of the Starling characteristics of the natural heart for each of the test conditions. The mock vasculature can be configured to mimic the input impedance and mechanical properties of native vasculature in the normal state. Although mock circulation testing systems cannot replace in vivo models, this configuration should be well suited for developing experimental protocols, testing device feedback control algorithms, investigating flow profiles, and training surgical staff on the operational procedures of cardiovascular devices. PMID- 14763491 TI - Myocardial hemodynamics, physiology, and perfusion with an axial flow left ventricular assist device in the calf. AB - The Jarvik 2000 axial flow left ventricular assist device (LVAD) is used clinically as a bridge to transplantation or as destination therapy in end-stage heart disease. The effect of the pump's continuous flow output on myocardial and end-organ blood flow has not been studied experimentally. To address this, the Jarvik 2000 pump was implanted in eight calves and then operated at speeds ranging from 8,000 to 12,000 rpm. Micromanometry, echocardiography, and blood oxygenation measurements were used to assess changes in hemodynamics, cardiac dimensions, and myocardial metabolism, respectively, at different speeds as compared with baseline (pump off, 0 rpm) in this experimental model. Microsphere studies were performed to assess the effects on heart, kidney, and brain perfusion at different speeds. The Jarvik 2000 pump unloaded the left ventricle and reduced end-diastolic pressures and left ventricular dimensions, particularly at higher pump speeds. The ratio of myocardial oxygen consumption to coronary blood flow and the ratio of subendocardial to subepicardial blood flow remained constant. Optimal adjustment of pump speed and volume status allowed opening of the aortic valve and contribution of the native left ventricle to cardiac output, even at the maximum pump speed. Neither brain nor kidney microcirculation was adversely affected at any pump speed. We conclude that the Jarvik 2000 pump adequately unloads the left ventricle without compromising myocardial metabolism or end-organ perfusion. PMID- 14763492 TI - Numerical simulation of a systemic flow test rig. AB - In this study, numerical simulation was carried out to investigate the dynamic response of a systemic flow test rig that is widely used for in vitro study of prosthesis in the cardiovascular system. In the system the physiological impedance of systemic circulation was modeled as a resistance-capacitance resistance type. The system analysis was directly based on differential equations describing the system dynamics, and numerically solved using the fourth-order Runge-Kutta method. Results showed that pressure in the systemic circulation test rig could be successfully simulated with the developed model. From the numerical experiment, it was found that the maximum stroke of the driving mechanism, the flow coefficients and opening of the control valves, and the initial volume of air in the compliance strongly affect the dynamic performance of the test rig. The numerical method developed is a useful tool in the design and optimization of the system configuration. PMID- 14763493 TI - Critically low hormone and catecholamine concentrations in the primed extracorporeal life support circuit. AB - The first hours of extracorporeal life support (ECLS) are commonly marked by new hemodynamic instability without a known etiology. We measured hormone and catecholamine concentrations in six ECLS primed circuits immediately before joining the patient's circulation to assess a potential role of these agents in this condition. The following hormones were significantly below the lower end of the normal range for the first week of life (data are presented as mean +/- SEM): cortisol 1.95 +/- 0.15 microg/dl (p < 0.001), aldosterone 3.73 +/- 0.74 ng/dl (p < 0.05), free thyroxine 1.2 +/- 0.1 ng/dl (p < 0.05), free triiodothyronine 0.53 +/- 0.03 pg/ml (p < 0.001), thyroid stimulating hormone 0.31 +/- 0.05 microU/ml (p < 0.001), growth hormone (GH) 0.09 +/- 0.01 ng/ml (p < 0.001), estradiol 38.3 +/- 3.72 pg/ml (p < 0.001), IGF-BP1 0.95 +/- 0.1 ng/ml (p < 0.001), glucagon 26 +/- 1.2 pg/ml (p < 0.001), epinephrine 17.3 +/- 3.7 pg/ml (p < 0.001), and norepinephrine 127 +/- 27 pg/ml (p < 0.05). No dopamine was detected. Normal hormone concentrations included IGF-I, IGF-BP3, insulin, parathyroid hormone, leptin, and testosterone. Critically low concentrations of cortisol, thyroid hormones, GH, IGF-BP1, glucagon and catecholamines were measured in the ECLS circuit even though it was primed with fresh frozen plasma. These concentrations may cause significant and precipitous dilutional reductions in the patient's circulating levels immediately after connection to the ECLS circuit and hence contribute to hemodynamic instability. PMID- 14763494 TI - Comparison of static airway pressures during total liquid ventilation while applying different expiratory modes and time patterns. AB - To compare pump driven (active) and gravity-siphon (passive) expiration modes during perfluorocarbon total liquid ventilation (TLV), a liquid ventilator was developed capable of providing either expiration mode. In a prospective, controlled laboratory study, 90 rabbits (3.2 +/- 0.1 kg) were anesthetized, tracheotomized, killed. After prefill with 12 ml/kg perflubron and TLV for 90 minutes (tidal volume 12 ml/kg, I:E ratio 1:2), randomly using passive (height 40 or 80 cm) or active expiration, respiratory rates were 4, 8, or 12/min. Static peak inspiratory and end-expiratory intratracheal pressures were measured at 5 minute intervals. Peak inspiratory and end-expiratory were constant in active groups, and increases in all 40 cm and 80 cm passive groups were significant. Differences between groups were significant for expiratory mode but not for respiratory rates. Only passive groups showed significant increases in body weight after TLV. Percentage of fluorothoraces was 10% using active and 85% using passive expiration. Based upon the stability of intrapulmonary pressures and volumes and a reduced rate of fluorothoraces, active expiration is more efficient than passive drainage during TLV. PMID- 14763495 TI - Continuous venovenous hemofiltration without anticoagulation. AB - We conducted a prospective observational study to assess the efficacy of continuous venovenous hemofiltration (CVVH) with no anticoagulation. A standard anticoagulation protocol for CVVH, which prescribed no anticoagulation for patients at risk of bleeding, was applied to 48 critically ill patients treated with CVVH. Circuit life was prospectively observed, and the following data were obtained for each circuit: heparin use and dose, protamine use, daily prothrombin time-international normalized ratio, activated partial thromboplastin time, and platelet count. Out of 300 consecutive circuits, 143 (47.6%) received no anticoagulation, 31 (10.3%) received regional anticoagulation, and 126 received low dose heparin. No patients experienced bleeding complications secondary to CVVH. Platelet count was significantly lower in the no anticoagulation group (73 x 10(3)/microl) compared with the low dose heparin group (119 x 10(3)/microl) and the protamine group (104 x 10(3)/microl) (p < 0.01 for both comparisons). There was no significant difference in mean circuit life among the three groups (heparin, 20.9 hours; no anticoagulation, 19.3 hours; protamine, 21.2 hours; not significant). In conclusion, for a group of patients deemed to be at risk of bleeding, CVVH without anticoagulation achieved an acceptable circuit life, which was similar to that obtained in other patients with low dose heparin anticoagulation or regional anticoagulation with heparin/protamine. PMID- 14763496 TI - Improved removal of small proteins using continuous venovenous hemofiltration to treat acute renal failure. AB - Continuous venovenous hemodialysis (CVVHD) or hemofiltration conducted with pre- (CVVHpre) or post- (CVVHpost) dilution modes are recommended to treat patients with acute renal failure (ARF) and cardiovascular instability. The efficiency of the three techniques was compared in a study including 18 critically ill patients with ARF. Their mean age was 62.1 +/- 16.7 years, and their mean SAPS II score was 59.5 +/- 14.3. They were treated sequentially with the three techniques for periods of 24 hours each (randomized assignment to one technique the first 24 hours followed by the two others). The PRISMA device and M 100 (AN69S) membrane were used in all instances. Blood and replacement (or dialysis) flow rates were kept at 150 and 25 ml/min, respectively. Urea, creatinine, uric acid, inorganic phosphorus, beta2 microglobulin (beta2m), and retinol binding protein (RBP) were measured every 12 hours in plasma and in 12 hours filtrate collection for 3 days. The results are expressed as filtrate/mean plasma (F/P) ratio for the 12 hour period. Removal of small molecules was 16% higher using CVVHD and CVVHpost than CVVHpre. For beta2m and RBP, CVVHpre was, respectively, 43% and 26% more efficient than CVVHD. CVVHpost gave higher but statistically different removal than CVVHpre only for beta2m. CVVHpost was the most efficient technique for removal of small proteins, but this advantage could be easily counterbalanced using higher volume substitution. PMID- 14763497 TI - Dialyzer performance in the HEMO Study: in vivo K0A and true blood flow determined from a model of cross-dialyzer urea extraction. AB - Inlet and outlet blood urea concentrations (Cin and Cout) can be used to directly measure dialyzer performance if simultaneous blood flow measurements (Qb) are available. Dialyzer clearance, for example, is the product of the urea extraction ratio [ER = (Cin - Cout)/Cin] and Qb. Urea concentrations are measured routinely in all hemodialysis clinics, but Qb is usually reported as the product of the pump rotational speed and pump segment stroke volume, which can be inaccurate at high flow rates. Dialyzer urea extraction is also a function of Qb, dialysate flow (Qd), and the membrane permeability-area coefficient (K0A) for urea. To determine true in vivo values for Qb and K0A in the absence of direct flow measurements, we developed a model based on an existing mathematical equation for hemodialyzer ER under conditions of countercurrent flow. Qb, K0A, and other variables were adjusted to fit the modeled ER to ER measured in 1,285 patients treated with Qb that ranged from 200 to 450 ml/min during the HEMO Study. Fitting was performed by least squares nonlinear regression using parametric and nonparametric methods for estimating true flow. As Qb rose above 250 ml/min, both methods for estimating actual Qb showed increasing deviations from the flow reported by the blood pump meter. Modeled values for K0A differed significantly among dialyzer models, ranging from 71% to 96% of the in vitro values. The previously described 14% increase in K0A, as Qd increased in vitro from 500 to 800 ml/min, was much less in vivo, averaging only 5.5 +/- 1.5% higher. Dialyzer reprocessing was associated with a 6.3 +/- 1.0% reduction in K0A and an approximate 2% fall in urea clearance per 10 reuses (p < 0.001). Multiple regression analysis showed a small but significant dialysis center effect on ER but no independent effects of other variables, including the ultrafiltration rate, diabetic status, race, ethnicity, sex, method of reuse, treatment time, access recirculation, and use of central venous accesses. The new algorithm allowed a more accurate determination of true Qb and in vivo K0A in the absence of direct flow measurements in a large population treated with a wide range of blood flow rates. Application of this technique for more than 1000 patients in the HEMO Study confirmed that in vitro measurements using simple crystalloid solutions cannot readily substitute for in vivo measurements of dialyzer function, and permitted a more accurate calculation of each patient's prescribed dialysis dose and urea volume. PMID- 14763498 TI - Blood flow and recirculation rates in tunneled hemodialysis catheters. AB - Long-term catheters are widely used in some dialysis units. Because of higher dialysis dose targets, high flow catheters have been made available. We measured blood flow (Qb in ml/min) and recirculation rate (R%) in two types of tunneled dialysis catheters using ultrasound-dilution technology (Transonic). Thirty-seven catheters were evaluated (27 Opti-Flow, 10 High-Flow), as inserted in jugular or subclavian veins. Real Qb and R were measured at increasing pump blood flows (250, 300, 350, 400, and 450 ml/min) in absence of ultrafiltration. For all, real Qb was similar to pump Qb (261 vs. 250, 304 vs. 300, 349 vs. 350, 389 vs. 400, and 431 vs. 450 ml/min, respectively). Catheters with reversed lines were all recirculating (R between 18% and 24%). Sixteen nonreversed catheters had no R at all Qb, whereas four nonreversed catheters had minimal R (between 7% and 11%); R did not increase significantly with the rise in pump Qb. The two types of tunneled catheters deliver high Qb without high R if ports are not reversed. The relative decrease in treatment efficiency should be accounted for in dialysis prescription if such tunneled catheters are used as long-term access, especially if lines have to be reversed. PMID- 14763499 TI - Extended reuse of polysulfone hemodialysis membranes using citric acid and heat. AB - The concomitant use of citric acid and prolonged exposure to heat (CAH) is an increasingly common alternative to purely chemical means of reusing dialyzers. However, there are no data on the effects of reprocessing dialyzers with CAH beyond 15 uses. Increasing the number of reuses with CAH cannot be systematically undertaken unless its safety is documented. We hypothesized that discarding polysulfone dialyzers after the 25th rather than the 15th use would result in increased clearance of beta2-microglobulin (beta2MG) without clinically significant changes in small solute clearance or albumin loss. We studied 15 Fresenius F80B polysulfone dialyzers in five chronic hemodialysis patients. Dialyzers were reprocessed using 1.5% citric acid solution heated to 95 degrees C. Representative fractional collection and 10 minute timed collections of dialysate were performed at baseline and during uses 5, 10, 15, 20, and 25 for each dialyzer. Dialysate-side urea, creatinine, and beta2MG clearances were calculated, and total albumin was measured in dialysate. We used a mixed model to adjust for repeated measures (both within a given dialyzer and for the multiple dialyzers per patient). Of the 15 dialyzers studied, 3 (20%) failed before the 25th use. There was no significant change in urea or creatinine clearance with additional reuse (overall p values 0.20 and 0.60, respectively). A sustained increase in beta2MG clearance was observed after the fifth treatment compared with the first use (p < 0.001). Fractional collection showed that dialysate albumin loss increased significantly with additional reuses (p < 0.001) but did not increase significantly above baseline until treatment 25. Reprocessing of polysulfone dialyzers with CAH 25 times significantly increased albumin loss and beta2MG clearance but did not appear to affect urea or creatinine clearance. Increasing the maximum number of uses to 20 may permit cost savings compared with current practice without additional risk. PMID- 14763501 TI - Cell Transplantation: The Regenerative Medicine Journal. A biennial analysis of publications. PMID- 14763500 TI - Impact of high volume hemofiltration on hemodynamic disturbance and outcome during septic shock. AB - The purpose of this study was to evaluate the effect of high volume continuous venovenous hemofiltration (HVCVVH) on hemodynamic and outcome in patients with septic shock. The primary end point was mortality at 28 days. Study design was a prospective case series, and study setting was a 12 bed intensive care unit at a university hospital. A total of 24 consecutive patients with septic shock were included, with dysfunction of more than two organs. All patients were treated by HVCVVH with ultrafiltration rate between 40 ml x kg(-1) x hr(-1) and 60 ml x kg( 1) x hr(-1) for 96 hours. In all patients, the increase in hemodynamic parameters was statistically significant (p < 0.05), with a significant linear decrease in norepinephrine doses (p < 0.05). The predicted 28 day mortality by three different severity scores was more than 70%, and the mortality in the hemofiltration group was 46% (p < 0.075). In the present study of septic shock patients with organ dysfunction, the hemodynamic parameters increased regularly during treatment by HVCVVH. This study suggests a beneficial effect of HVCVVH on 28 day mortality (46% vs. 70%), and further studies with larger cohorts are required. PMID- 14763502 TI - Neural precursor cells as carriers for a gene therapeutical approach in tumor therapy. AB - Conventional therapeutical approaches such as surgery, radiotherapy, or chemotherapy have been shown to be rather unsuccessful in the treatment of infiltrative growing tumors such as the malignant glioblastoma multiforme. Thus, new therapeutical strategies have to be developed that are suitable for inducing cell death also in migrating tumor cells. These new therapeutical stategies include cell and/or gene therapeutical approaches. We demonstrate that glial restricted progenitor cells as well as embryonic stem cell-derived neural stem cells belong to cell populations applicable to such therapeutical concepts. Both cell types can be efficiently transduced using a third-generation high-capacity "gutless" adenoviral vector, and show a tropism for the F98 glioma cells by migrating towards a spheroid of F98 glioma cells with a tendency to form a barrier around the tumor spheroid in an in vitro tumor confrontation model. Moreover, in a migration assay, secretion products of glial-restricted precursor cells have shown a potency to inhibit the migratory activity of glioma cells in vitro. In vivo, F98 glioma cell-derived tumor formation in the right striatum resulted in migration of glial as well as neural precursor cells towards the tumor area when cotransplanted in the corpus callosum of the contralateral hemisphere. After arrival, both cell types surround the tumor mass and even invade the experimentally induced tumor. These data indicate that glial restricted as well as embryonic stem cell-derived neural precursor cells are good candidates as carriers for an ex vivo gene therapeutical approach in tumor therapy. PMID- 14763503 TI - Expression of neuronal markers in differentiated marrow stromal cells and CD133+ stem-like cells. AB - Bone marrow stromal cells, which normally give rise to bone, cartilage, adipose tissue, and hematopoiesis-supporting cells, have been shown to differentiate in vitro and in vivo into neural-like cells. In this study, we examined the expression of neuronal and glial markers in human marrow stromal cells under culture conditions appropriate for neural stem cells, and compared the unsorted cell population to bone marrow CD133+ stem-like cells using immunofluorescence, Western blot, and functional patch-clamp analysis. Overall, the expression of the early neuronal marker beta3-tubulin was most pronounced in the presence of DMEM/F12 and neurotrophin 3 (NT3) or brain-derived neurotrophic factor (BDNF), when marrow stromal cells were cultured onto fibronectin. Electrophysiological examination, however, could not show fast sodium currents or functional neurotransmitter receptors in differentiated marrow stromal cells. CD133+ mesenchymal stem-like cells, but not CD34+/CD133- cells, generally showed a higher expression of neuronal markers than did unsorted marrow stromal cells, and differentiated CD133+ cells more resembled neuron-like cells. PMID- 14763504 TI - Insulin-induced normoglycemia reduces islet number needed to achieve normoglycemia after allogeneic islet transplantation in diabetic mice. AB - The Edmonton protocol established that insulin independence could be reached with the transplantation of an appropriate number of islet cells. However, to effect a cure, islets from two or three pancreases are needed. The aim of this study was to examine whether normoglycemia, with insulin treatment before and after transplantation, reduces the islet number needed to achieve normoglycemia in allogeneic islet transplantation. Swiss mice were used as donors and recipients. Diabetes was induced by i.p. administration of streptozotocin (180 mg/kg BW). Diabetic mice were transplanted with 300 (n = 16), 400 (n = 16), or 500 (n = 16) islets under the left kidney capsule. For every group, half the animals were kept normoglycemic with insulin treatment from day 4 before transplantation to day 10 after transplantation. At the end of the study, all normoglycemic mice were given an i.p. glucose tolerance test (IPGTT). For statistical analysis, paired or unpaired Student's t-test or ANOVA was used. Only insulin-treated mice achieved normoglycemia by the end of the study (37.5% of animals transplanted with 400 islets and 50% transplanted with 300 or 500 islets). At the end of the study, normoglycemic mice transplanted with 300 allogeneic islets showed better glycosylated hemoglobin (HbA1C) than did normoglycemic mice transplanted with 500 islets (300 islets: 2.7 +/- 0.2%; 500 islets: 3.6 +/- 0.2%; p < 0.05). After the IPGTT, insulin-treated mice transplanted with 500 islets showed abnormal glucose tolerance; however, insulin-treated mice transplanted with 300 or 400 islets showed normal glucose tolerance. Insulin treatment reduced the islet number needed to achieve normoglycemia in allogeneic islet transplantation. The HbA1C and IPGTT results suggest that transplanting smaller numbers of allogeneic islets improves beta-cell function; some studies suggest that this may be due to lower immunogenicity, hypoxia, and inflammation. PMID- 14763505 TI - Hyperthermic preconditioning protects pig islet grafts from early inflammation but enhances rejection in immunocompetent mice. AB - The induction of heat shock proteins (HSP) protects isolated islet cells against the cytotoxicity of inflammatory mediators in vitro. Very little information is available about the effect of HSP overexpression on function of preconditioned islet grafts. The present study investigated the function of heat-exposed pig islets after transplantation into immunocompetent mice in comparison with in vitro resistance against inflammatory mediators. Pig islets were preconditioned at 43 degrees C or sham treated prior to subcapsular transplantation into diabetic C57/Bl6j mice. Nondiabetic mice simultaneously receiving preconditioned and control islets were subjected to bilateral nephrectomy for determination of pig insulin. Resistance against H2O2, NO, human Il-1beta, IFN-gamma, or TNF-alpha was assessed by trypan blue exclusion and insulin determination. Heat-induced protein expression was confirmed by Western blot analysis. Graft preconditioning increased resistance against H2O2, NO, or cytokines (p < 0.05) but decreased survival in nondiabetic mice (p < 0.05) and function in diabetic mice (p < 0.01). Upregulation of caspase-3 activity as well as Bax, Fas, FasL, and DFF expression (p < 0.05) indicated simultaneous induction of apoptosis. The coexpression of HSP and proapoptotic proteins reveals the dual character of the stress response simultaneously starting mechanisms for protection and apoptosis. In vitro assays seem to reflect only insufficiently the situation of islets after transplantation. PMID- 14763507 TI - Improvement of pancreatic islet isolation outcomes using glutamine perfusion during isolation procedure. AB - During procurement, isolation, and transplantation, islets are exposed to high levels of oxidative stress triggering a variety of signaling pathways that can ultimately lead to cell death. Glutamine is an important cellular fuel and an essential precursor for the antioxidant glutathione. The aim of this study was to examine the role of intraductal glutamine administration in facilitating recovery of isolated rat islets from pancreases subjected to a clinically relevant period of warm ischemia. Islets were isolated in Sprague-Dawley (SD) rats (n = 18 per group). Pancreata in groups 1 and 2 were procured immediately while groups 3 and 4 were subjected to 30-min warm ischemia. Groups 2 and 4 were treated intraductally with 5 mM glutamine prior to pancreatectomy. Exposure to 30-min warm ischemia significantly reduced islet yield [groups 1 & 2 (nonischemia): 503 +/- 29 islets/rat vs. groups 3 & 4 (ischemia): 247 +/- 26 islets/rat; p < 0.05]. Intraductal glutamine treatment significantly improved islet yield when pancreata were subjected to 30-min warm ischemia [144 +/- 16 islets/rat without glutamine (group 3) vs. 343 +/- 36 islets/rat with glutamine (group 4), p < 0.05]. Glutamine also significantly improved islet viability (values were 50 +/- 4% in group 4 vs. 27 +/- 3% in group 3, p < 0.05). Similarly, glutathione (reduced) levels were significantly elevated in both glutamine-treated groups; however, this increase was greatest in tissues exposed to ischemia (2.76 +/- 0.04 nmol/mg protein in group 4 vs. 1.66 +/- 0.04 nmol/mg protein in group 3, p < 0.05). Intraductal glutamine administration considerably improves the islet yield, viability, and augments endogenous glutathione levels in pancreata procured after a clinically relevant period of ischemia. Intraductal administration of glutamine at the time of digestive enzyme delivery into the harvested pancreas may represent a simple yet effective tool to improve islet yields in clinical isolations. PMID- 14763506 TI - Response of encapsulated rat pancreatic islets to hypoxia. AB - Hypoxia contributes to encapsulated pancreatic islet graft failure. To gain insight into the mechanisms that lead to hypoxia-induced graft failure, encapsulated islet function, vitality, and cell replication were assessed after 2 and 5 days of hypoxic (1% O2) and normoxic (20% O2) culture. The mRNA expression levels of Bcl-2, Bax, inducible nitric oxide synthase (iNOS), and monocyte chemoattractant protein 1 (MCP-1) were assessed, as well as the amount of nitrite and MCP-1 in the culture medium. Hypoxia was associated with loss of encapsulated islet function and vitality, but not with an increase in islet cell replication. Loss of vitality was due to necrosis, and only modestly due to apoptosis. Hypoxia was not associated with changes in the Bcl-2/Bax mRNA ratio, but it did increase the expression of iNOS and MCP-1 mRNA. The increased mRNA levels were, however, not associated with elevated concentrations of nitrite nor with elevated levels of MCP-1 protein. The increased iNOS mRNA levels imply a role for NO in the completion of cell death by hypoxia. The increased MCP-1 mRNA levels suggest that encapsulated islets in vivo contribute to their own graft failure by attracting cytokine-producing macrophages. The discrepancy between iNOS mRNA and nitrite is explained by the longer half-life of NO during hypoxia. MCP-1 protein levels are underestimated as a consequence of the lower number of vital cells in combination with a higher proteolytic activity due to necrosis. Thus, strategies to eliminate hypoxia may not only improve islet function and vitality, but may also reduce the attraction of macrophages by encapsulated islets. PMID- 14763508 TI - A simple and cost-effective method for the isolation of islets from nonhuman primates. AB - Recent advances in islet cell transplantation have led to insulin independence in a majority of islet transplant recipients. However, there exists a need to overcome the shortage of donor tissue and the necessity for life-long immunosuppression. Preclinical studies in large animal models are necessary to evaluate the safety and efficacy of alternative approaches for clinical islet transplantation. The nonhuman primate serves as an appropriate animal model for such investigations; however, a major impediment in performing such preclinical research has been the difficulty in isolating islets of sufficient quantity and quality. The current study describes a simple and cost-effective method to isolate nonhuman primate islets to support preclinical islet transplantation research. The results of islet isolations from 54 cynomolgus monkeys and 4 baboons are reported. The pancreas was infused with Liberase HI and subjected to static digestion. The digested tissue was shaken, filtered through a mesh screen, applied to a discontinuous gradient, and centrifuged in much the same manner as with conventional rodent islet isolations. Islets were collected from the two interfaces, washed, and transplanted. Following purification, cynomolgus monkey islet isolation yields were 50,100 +/- 3120 IE total or 8760 +/- 420 IE/g pancreas with the percent purity and viability of 90.8 +/- 0.9 and 90.7 +/- 0.7, respectively. Total insulin content of the isolated islets was 405 +/- 53 microg insulin with DNA content being and 976 +/- 117 microg DNA, corresponding to a ratio of 0.57 microg insulin/microg DNA. STZ-induced diabetes was reversed in both mouse and nonhuman primate recipients, which possessed significant levels of c-peptide following transplantation and well-granulated islet grafts. The technique yields sufficient numbers of pure and viable islets to support preclinical research to develop improved strategies to prevent the immune destruction of the transplanted islet graft. PMID- 14763509 TI - Hepatic cells via cava vein can influence allogenic islet rat transplantation. AB - We have reported, previously, some effect of allogenic hepatic cells for islet tolerance when they are injected mixed (hepatic cells and islets) in different proportions via portal vein, in diabetic Wistar rats. Now we have studied the role of allogenic hepatic cells injected sequentially 15 min before islets, comparing via the portal vein (A and B groups) and via the cava vein (C and D groups) with a control group of islets alone. The allogenic islets were always injected via portal vein, in similar conditions, while the ratio of hepatic cells/islets was 100:1 (A, C groups) or 200:1 (B, D groups). Islets and hepatic cells were obtained from several different rats. The transplanted rats were observed during 30 days and results compared among the different rat groups: porta-porta (P/P), cava-porta (C/P), and control group. Statistically, a significant interaction between type of transplant and proportion of hepatic cells was observed. Also, C plus D groups showed statistical difference with the control group (p < 0.017) and also all the groups together (p < 0.047). These results suggest that hepatic cells can induce, in some cases, islet graft prolongation in Wistar rats. Better results were obtained when hepatic cells were injected via the cava vein than via the portal vein. Because we used a liver cell suspension integrated for several kinds of cells, the study does not clarify if this effect can be related to some specific hepatic cell subpopulation. To confirm the results and to determine if the hypothetical mechanism can be attributed to a block of the immune system or to some factor secreted by hepatic cells, more studies must be performed. PMID- 14763510 TI - In vitro expansion of human hepatocytes is restricted by telomere-dependent replicative aging. AB - Currently, different techniques to expand human hepatocytes in vitro are being investigated to generate enough cells for liver-directed cell therapies. However, based on observations in fibroblasts and other cell types, telomere attrition limits the proliferative capacity of normal somatic cells. Therefore, we explored whether telomere-dependent replicative aging restricts the in vitro proliferation of human hepatocytes. Subpopulations of cells isolated from a neonatal liver and characterized as hepatocyte derived by RT-PCR and flow cytometry started to proliferate 5-7 days after plating and were termed proliferating human hepatocytes (PHH). Following retroviral-mediated transduction of the catalytic telomerase subunit, telomerase reverse transcriptase (hTERT), telomerase activity increased from almost undetectable levels to levels as high as in HepG2 and other telomerase-positive cell lines. As expected, untransduced PHH progressively lost telomeric repeats and arrested after 30-35 cell divisions with telomeres of less than 5 kilo bases. In comparison, telomerase-reconstituted PHH maintained elongated telomeres and continued to proliferate as shown by colorimetric assays and cell counts. In this study, telomere stabilization extended the proliferative capacity of in vitro proliferating human neonatal hepatocytes. Therefore, telomere attrition needs to be addressed when developing techniques to expand human hepatocytes. PMID- 14763511 TI - p27Kip1 inactivation provides a proliferative advantage to transplanted hepatocytes in DPPIV/Rag2 double knockout mice after repeated host liver injury. AB - Studies were conducted to develop a new DPPIV(-/-)/Rag2(-/-) mouse model for hepatocyte transplantation by allogeneic and xenogeneic cells and to compare the proliferative capacity of p27 null hepatocytes versus normal hepatocytes in this system. Dipeptidyl peptidase IV (DPPIV) gene knockout mice, in which wild-type (wt) DPPIV+ donor hepatocytes can be readily identified by enzyme histochemistry, were bred with Rag2 null mice to prepare immunotolerant DPPIV(-/-)/Rag2(-/-) double knockout mice. DPPIV(-/-)/Rag(-/-) mice were transplanted with wt hepatocytes or p27 null mouse hepatocytes, which show enhanced cell cycle activity due to disruption of the Kip1 cyclin kinase inhibitor gene, and liver repopulation was assessed under nonproliferative versus proliferative experimental conditions. After their initial engraftment, transplanted wt hepatocytes did not proliferate in untreated livers or increase significantly in response to an acute liver regenerative stimulus. p27 null hepatocytes engrafted with the same efficiency as wt hepatocytes, but showed a noticeable, although not statistically significant, increase in proliferation in response to partial hepatectomy or acute CCl4 administration. Repeated treatments with CCl4 substantially increased proliferation and liver repopulation by p27 null hepatocytes but not by wt hepatocytes. These results suggest that p27 gene inactivation does not overcome proliferative restrictions imposed on hepatocytes by the normal liver, but that after repeated episodes of toxic liver injury, the augmented proliferative capacity of p27 null hepatocytes leads to significant liver repopulation compared with wt hepatocytes. These properties of p27 deficient hepatocytes could prove useful as a target for liver repopulation in patients with intermittent or a low level of chronic liver injury. PMID- 14763512 TI - Detection of Streptococcus anginosus and 8-hydroxydeoxyguanosine in saliva. AB - Several studies have demonstrated a close association between Streptococcus (S.) anginosus infection and head and neck cancer. Accumulation of 8-hydroxy deoxyguanosine (8-OHdG), which may result from the continuous generation of reactive oxygen species associated with chronic inflammation, has been reported in human preneoplastic lesions and in cancerous tissues. The purpose of the present investigation was to assess the salivary levels of S. anginosus and 8 OHdG in patients with periodontitis. Salivary levels of S. anginosus were measured by real-time PCR. S. anginosus was detected in 28 out of 38 (73.7%) of subjects. The 8-OHdG level was significantly higher in patients positive for S. anginosus than in patients negative for the bacterium. A significant decrease in S. anginosus and 8-OHdG levels was observed after initial periodontal treatment. Our findings indicate that, although the levels of S. anginosus are relatively low, there is a correlation between the salivary level of S. anginosus and 8 OHdG, and that periodontal treatment can decrease the levels of these hazard factors. PMID- 14763513 TI - GABA(A) receptors in the nucleus accumbens core modulate turning behavior induced by dopamine receptor stimulation. AB - The role of GABA(A) and GABA(B) receptors in the core of the nucleus accumbens in turning behavior of rats was investigated. Unilateral injections into the core of the nucleus accumbens of the GABA(A) receptor agonist (muscimol, 50 ng) and antagonist (bicuculline, 200 ng), and the GABA(B) receptor agonist (baclofen, 100 ng) and antagonist (2-hydroxysaclofen, 2 microg) did not produce turning behavior. In rats pretreated with unilateral injections of the dopamine D1 like/D2-like receptor antagonist, cis(Z)-flupentixol (10 microg), into the ventrolateral striatum and saline into the nucleus accumbens core of contralateral side, systemic injection of a mixture of dopamine D1-like (SKF 38393, 3 mg/kg) and D2-like (quinpirole, 1 mg/kg) receptor agonists has been found to elicit contraversive pivoting, namely pivoting away from the side of the core injection. This dopamine D1-like/D2-like receptor-mediated pivoting was significantly inhibited by injections into the core of the nucleus accumbens of muscimol (50 ng), but not bicuculline (200 ng). In contrast, the dopamine D1 like/D2-like receptor-mediated pivoting was suppressed by either baclofen (100 ng) or 2-hydroxysaclofen (2 microg) injected into the nucleus accumbens core. It is therefore concluded that neither GABA(A) nor GABA(B) receptor stimulation in the core of the nucleus accumbens produces turning behavior, and that GABA(A), but not GABA(B), receptors in the nucleus accumbens core may modulate dopamine D1 like/D2-like receptor-mediated pivoting. PMID- 14763514 TI - Fluoride availability and stability of Japanese dentifrices. AB - The decline of dental caries in the industrialized countries can be attributed to widespread use of fluorides. The Japanese market share of dentifrices containing fluorides has increased from 12% in 1985 to 77% in 2000. For a fluoride dentifrice to be effective in the control of dental caries, an adequate concentration of the fluoride must be soluble. Several Japanese fluoride dentifrices have in their formulations calcium phosphate as an abrasive, which may react with fluoride. This study was designed to evaluate the availability and stability of fluoride in the most consumed dentifrices in Japan. The analyses were made when the dentifrices were purchased (fresh samples) and after one year of storage (aged samples) at room temperature (21.8 +/- 3.6 degrees C). Total fluoride and soluble fluoride was determined using an ion specific electrode. All dentifrices showed similar concentrations of total fluoride in fresh and aged samples in accordance with the Japanese Legislation (content of less than 1,000 ppm F). Some dentifrices, with dicalcium phosphate as abrasive, showed decreasing concentrations of total soluble fluoride and increasing amounts of insoluble fluoride over time. Although most of the Japanese fluoridated dentifrices evaluated in this study contain unstable fluoride, they were found to have sufficient concentrations of soluble fluoride to be effective in preventing dental caries. PMID- 14763515 TI - The anti-adherence effect of Piper betle and Psidium guajava extracts on the adhesion of early settlers in dental plaque to saliva-coated glass surfaces. AB - The aqueous extracts of Piper betle and Psidium guajava were prepared and tested for their anti-adherence effect on the adhesion of early plaque settlers (Strep. mitis, Strep. sanguinis and Actinomyces sp.). The saliva-coated glass surfaces were used to simulate the pellicle-coated enamel surface in the oral cavity. Our results showed that the anti-adherence activities of Piper betle and Psidium guajava extracts towards the bacteria were different between the bacterial species. Psidium guajava was shown to have a slightly greater anti-adherence effect on Strep. sanguinis by 5.5% and Actinomyces sp. by 10% and a significantly higher effect on Strep. mitis (70%) compared to Piper betle. The three bacterial species are known to be highly hydrophobic, and that hydrophobic bonding seemed to be an important factor in their adherence activities. It is therefore suggested that the plant extracts, in expressing their anti-adherence activities, could have altered the hydrophobic nature of the bonding between the bacteria and the saliva-coated glass surfaces. PMID- 14763516 TI - Investigation of the bone tissue response to glass-ionomer microimplants in the canine maxillary alveolar ridge. AB - This paper reports the results of experimental use of glass-ionomer microimplants in the augmentation of the maxillary alveolar ridge in dogs. The study included ten adult mongrel dogs 5 years of age, weighing between 50 and 70 pounds (25-30 kg), divided into 2 groups of 5 dogs each. In both groups, the maxillary 4th premolar and 1st molar were removed after the elevation of a buccal mucoperiosteal flap. The alveolar bone adjacent to the extracted teeth was also removed. In the experimental group (5 dogs), Ionogran a glass-ionomer microimplants (GIMIs) (Ionogran particle size of 0.5-1.0 mm, IONOS Medizinische Produkte GmbH & Co. KG, D-8031 Seefeld, Gemany) were used for augmentation and were inserted in the created defects. The extraction sockets and bone defects were augmented with an average amount of 2 g of GIMIs. In the control group, the bone defects were left unfilled as a control for bone healing. Histological examination showed that the glass-ionomer microimplants were extremely osteoconductive and inert materials. Stimulation of growth of new bone tissue in contact with the glass-ionomer microimplants was evident. No inflammatory cells were detected on or adjacent to the GIMIs. In the control group, incomplete bone healing with fibrous scar tissue and inflammatory cells was noted. These results indicate that glass-ionomer microimplants represent highly osteoconductive and biocompatible materials for use in bone surgery. PMID- 14763517 TI - Histogenesis and disappearance of the teeth of the Mekong giant catfish, Pangasianodon gigas (Teleostei). AB - Juveniles of the Mekong giant catfish, Pangasianodon gigas (Teleostei), have 3 sorts of tooth-upper and lower jaw teeth, palatal teeth, and pharyngeal teeth- but adults are toothless. To investigate the histogenesis and disappearance of the teeth, we made serial sections of the mouth and teeth of juvenile fish at 10 developmental stages (from ca. 8.5 to ca. 30 cm in total length) and examined them under scanning electron microscope and light microscope. Observations of teeth and surrounding tissues in the serial sections revealed the process of tooth resorption by active odontoclast-like cells. Numbers of jaw and palatal teeth decreased with age. When the fish reached ca. 14 cm in total length, the numbers of functional upper jaw teeth and successional tooth germs decreased rapidly, and the developmental rate of successional tooth germs slowed. When the fish reached ca. 24 cm, no teeth existed in the upper jaw. It is clear that tooth disappearance results from the shedding of functional teeth and the lack of replacement tooth germs. PMID- 14763518 TI - Calcium pyrophosphate dihydrate arthropathy with condylar destruction of the temporomandibular joint. AB - We report a case of calcium pyrophosphate dihydrate (CPPD) arthropathy with condylar destruction of the temporomandibular joint. Diagnosis was made on the basis of clinical findings and x-ray evidence of severe destruction of the condyle. The most likely diagnosis was considered to be neoplasm and a secondary infection of the left TMJ. Review of sections of the condyle taken at operation confirmed the diagnosis of left TMJ pseudogout. The postoperative course was uneventful with improvement in the clinical symptoms. PMID- 14763519 TI - Treatment planning in a case of restoration of the maxilla and mandible using osseointegrated implants with four types of bone graft. AB - A case is reported of a 66-year-old woman who could not use a conventional, full upper denture because of a gag reflex. In the maxillary alveolar ridge, restoration was performed on a moderately atrophied, edentulous anterior area and a small defect in the right-side posterior area. In the mandibular alveolar ridge, restoration was performed on a moderate osseous defect in each molar area resulting from tooth extraction due to severe periodontal disease. Based on careful treatment planning, four types of bone graft were used with previously designed osseointegrated implants. The atrophied maxillary alveolar ridge was restored with veneer iliac bone grafts to avoid fenestration during implant placement, while alveolar process deficiency was restored using inlay and sinus bone grafts as placements for long implant fixtures. The defects in the mandibular alveolar bone were filled with corticocancellous bone chips at the implant placement sites. A combination of immediate and secondary placement of Branemark fixtures was used. Bone-anchored bridge-type implant prostheses were fitted approximately twelve months after surgery. Three years later, there had been no failure of implant fixtures and satisfactory functional and cosmetic restoration had been maintained. PMID- 14763520 TI - Is there evidence of a sphincter system in Wharton's duct? Etiological factors related to sialolith formation. AB - The exact cause of the formation of sialoliths is unknown. Detailed knowledge of the pathogenesis of sialolithiasis is necessary to define new therapeutic procedures. The possible presence of a sphincter system in Wharton's duct has been described frequently in the context of diagnostic sialendoscopy. Serial histological examination of the entire Wharton's duct in four samples revealed no anatomical correlation for the presence of a sphincter. Secretory disturbances and viscous secretions as well as microlith formation and ductal obstruction cannot fully explain the genesis of sialoliths. The coaction of those factors with participation of bacteria leads to the development of sialoliths. PMID- 14763522 TI - Intramedullary nailing. PMID- 14763521 TI - Hyperbaric oxygen use. PMID- 14763523 TI - Orthopedics and online manuscript submission. PMID- 14763524 TI - Radiologic case study. Gouty tophus involving the distal quadriceps tendon. AB - This case depicts an unusual presentation of gout, which could be mistaken for sarcoma. The radiographic and MRI findings suggest gout whereas the CT appearance is most specific. Therefore, CT should be considered when MRI demonstrates low to intermediate T2 signal in a heterogeneously enhancing soft-tissue mass around a joint, tendon, or bursa, especially when radiographs show adjacent erosion with a sclerotic margin. Gouty tophus is not excluded by a normal serum uric acid level. Computed tomography or ultrasound can be used to guide needle biopsy to provide diagnostic tissue. PMID- 14763525 TI - Modified Weaver-Dunn procedure for acromioclavicular joint dislocations. PMID- 14763526 TI - ACL reconstruction using quadriceps tendon. PMID- 14763527 TI - The octogenarian total knee arthroplasty. AB - Sixty-five patients aged > 80 years were compared to 65 patients aged between 60 and 70 years to assess total knee arthroplasty (TKA) outcome using the octogenarian postoperative Knee Society knee and function scores. Knee Society knee scores showed excellent outcomes with no statistically significant difference in the younger cohort (preoperative score, P = .7156; 5-year score, P = -.0677). Knee Society function scores also showed good outcomes with no statistically significant difference between the groups (preoperative score, P = .1147; 5-year score, P = .1348). Average length of stay increased by 3 days for octogenarians. Octogenarians had more pre-existing medical conditions and suffered more postoperative confusion. All but 1 patient rated the result as good/excellent. Patients maintained their independence for approximately 3.5 years before requiring more social input. With careful preoperative planning and counseling, TKA is recommended for the octogenarian. PMID- 14763528 TI - Incidence of heterotopic ossification following total hip arthroplasty in patients with prior stroke. AB - This study reviewed the incidence of heterotopic ossification and the functional limitations in a cohort of consecutive patients with prior stroke who underwent primary total hip arthroplasty (THA). Thirty-one primary THAs were performed in 22 patients who had a cerebrovascular accident prior to THA. Mean follow-up 35 months. The overall incidence of heterotopic ossification was 36%, with significant Brooker class III and IV heterotopic ossification reported in 22% of patients. The data suggest that prior cerebrovascular accident may pose an increased risk of significant heterotopic ossification following primary THA. Consideration of prophylaxis in this subset of patients may be warranted. PMID- 14763529 TI - Surgical treatment for cervical myelopathy in patients aged > 80 years. AB - The surgical outcomes of 13 patients who were diagnosed with cervical spondylotic myelopathy were reviewed retrospectively. Mean patient age at surgery was 83 years. The severity of cervical spondylotic myelopathy was evaluated using the Japanese Orthopaedic Association score. Daily activities were evaluated using the Barthel index. The preoperative JOA score and Barthel index were 7.8 and 63.5, respectively. The mean JOA score and Barthel index maximum recovery rate were 35% and 24%, respectively. The results of this study imply that surgery for patients with cervical spondylotic myelopathy aged > 80 years is warranted. PMID- 14763530 TI - Thoracoscopic sympathectomy in the management of vasomotor disturbances and complex regional pain syndrome of the hand. AB - Complex regional pain syndrome, vasospastic disorders, and hyperhidrosis are chronic and debilitating upper extremity problems. Twenty-nine consecutive patients treated with thoracoscopic sympathectomy are presented. Diagnoses included complex regional pain syndrome, hyperhidrosis, Buerger's disease, Raynaud's disease, and peripheral vascular disease. All patients with hyperhidrosis had complete symptom resolution. Patients with Buerger's and Raynaud's disease had excellent/good results. Six patients with complex regional pain syndrome had excellent or good relief; the remaining six patients had varying degrees of recurrence. A statistically significant association was noted between duration of complex regional pain syndrome prior to sympathectomy and outcome. Thoracoscopic sympathectomy is an effective treatment for hyperhidrosis and vasospastic disorders. Although the results for complex regional pain syndrome are not uniformly excellent, this technique offers promise in the treatment of this difficult problem. PMID- 14763531 TI - Evaluation of a new power-operated PMMA vacuum mixing and delivery system for cemented femoral stem insertion. AB - Variations exist in the manual preparationand delivery of cement during primary total hip arthroplasty. The incorporation of a power source may standardize cement mantle manufacturing. This study prospectively compared the use of a new power-operated polymethylmethacrylate vacuum mixing and delivery system to a commonly used manually powered industry standard. The study did not reveal a significant difference in the quality of cement mantle grades when comparing the use of this power-operated system to an industry standard manual system. However, the benefit of the new preparation and delivery system may be in consistent porosity reduction and not in the improvement of cement mantle grading. PMID- 14763532 TI - Hybrid thin wire external fixation: an effective, minimally invasive, modular surgical tool for the stabilization of periarticular fractures. AB - Hybrid thin wire external fixation is an effective, minimally invasive treatment for the stabilization of periarticular, supracondylar, and pilon fractures. The extent of bone and soft-tissue loss, high risk of infection, and further damage to the soft tissues precludes open reduction and internal fixation as a safe treatment method. External fixation preserves the soft-tissue envelope with minimal damage and allows fracture stabilization, early loading, and mobilization, which promote bone healing. PMID- 14763533 TI - Femoral chondrosarcoma complicating Paget's disease of bone. PMID- 14763534 TI - Injection-induced contracture of the quadriceps femoris muscle in children. PMID- 14763535 TI - Giant-cell reparative granuloma of the hands and feet. PMID- 14763536 TI - Traumatic triple tendon rupture in secondary hyperparathyroidism. PMID- 14763537 TI - Current overview of neurovascular structures in hip arthroplasty: anatomy, preoperative evaluation, approaches, and operative techniques to avoid complications. AB - Total hip arthroplasty is a common and relatively safe procedure with consistently good results. Despite its popularity and excellent results, THA is a major operation with several major neurovascular structures within reach of retractors, scalpel blades, drills, screws, and reamers. A thorough knowledge of their anatomic location and proximity to the operative field, along with a basic understanding of the principles of vascular surgery can help avoid potentially devastating consequences. Specifically, the surgeon should avoid placement of screws in the anterior-superior quadrant, be vigilant when placing retractors, and avoid excessive tension on the sciatic nerve. PMID- 14763538 TI - The use of an injectable bone graft substitute in tibial metaphyseal fractures. AB - Eight consecutive patients with severely comminuted proximal or distal tibial metaphyseal fractures were surgically treated with a novel injectable, in situ hardening calcium sulfate material to provide temporary intraoperative structural support until instrumentation was appropriately placed. The calcium sulfate bone substitute was also used as a bone void filler to provide an osteoconductive environment following stabilization. Bone regrowth was observed in all patients and, in seven of the eight patients, 90%-100% bone formation was observed within 3 months. The bone substitute had almost completely resorbed by 3 months in all patients. One patient with extensive initial bone loss required a second bone graft. Although the patient sample size was small, the success rate of treating severely comminuted fractures was significantly better than in reports using only internal fixation. Injectable bone substitute for minimally invasive surgery to repair metaphyseal fractures. PMID- 14763539 TI - Successful treatment of a persistent humeral shaft nonunion using open reduction internal fixation, electrical stimulation, and AlloMatrix Custom Bone Putty. PMID- 14763540 TI - Effects of altered crystalline structure and increased initial compressive strength of calcium sulfate bone graft substitute pellets on new bone formation. AB - A new, modified calcium sulfate has been developed with a different crystalline structure and a compressive strength similar to many calcium phosphate materials, but with a resorption profile only slightly slower than conventional surgical grade calcium sulfate. A canine bilateral defect model was used to compare restoration of defects treated with the modified calcium sulfate compared to treatment using conventional calcium sulfate pellets after 6, 13, and 26 weeks. The modified calcium sulfate pellets were as effective as conventional calcium sulfate pellets with regard to the area fraction and compressive strength of newly formed bone in the treated bone defects. Mechanical testing demonstrated that the initial compressive strength of the modified material was increased nearly three-fold compared to that of conventional surgical-grade calcium sulfate. This increase potentially allows for its use in a broader range of clinical applications, such as vertebral and subchondral defects. PMID- 14763541 TI - Evaluation of a synthetic bone defect test model to aid in the selection of materials for use in vertebral body compression fracture repair. AB - A synthetic test model was developed to assist in screening injectable cements with a focus on mechanical strength for vertebral body compression fracture repair. The two-part defect model consisted of a polyurethane foam cube to simulate trabecular bone and a defect to which various injectable cements could be introduced. In addition, a finite element analysis model was developed and the results were compared to laboratory testing. Agreement was found between the finite element analysis and test results. Once the finite element analysis model was validated with experimental data, an additional finite element analysis was conducted to study various parameters affecting mechanical performance such as simulated bone and cement stiffness. Finite element analysis models were also created using orthotropic bone properties typical of healthy trabecular bone and were compared to various foam stiffnesses. The foam model was a good in vitro representation of actual trabecular bone found in vertebral bodies and is a valid model to evaluate the mechanical strength of injectable cements for percutaneous vertebral body fracture repair. PMID- 14763542 TI - Use of cancellous bone chips and demineralized bone matrix in the treatment of acetabular osteolysis: preliminary 2-year follow-up. AB - Twenty patients (20 hips) who had cementless acetabular revision arthroplasty and were treated with cancellous bone chips mixed with demineralized bone matrix to fill the cavitary defects were studied. At 2-years' follow-up, the grafts were fully incorporated in 18 patients. In addition, the mean Harris Hip Score increased to 89 points from a mean preoperative score of 29 points for the surviving hips. These preliminary findings suggest that the prepackaged demineralized bone matrix and cancellous chips can provide acceptable radiographic and clinical results as an adjunct in the treatment of contained osteolytic acetabular defects. PMID- 14763543 TI - Repair of a severely comminuted distal femur fracture using surgical-grade calcium sulfate bone graft substitute. PMID- 14763544 TI - Minimally invasive treatment of non-steroid induced knee osteonecrosis of the lateral femoral condyle. PMID- 14763545 TI - Treatment of benign bone lesions with an injectable calcium sulfate-based bone graft substitute. AB - The treatment of benign bone lesions can be challenging due to the limited quantity of autogenous graft available for harvest and grafting of the defect. The use of an injectable calcium bone graft substitute, Minimally-Invasive Injectable Graft (MIIG) (Wright Medical Technology, Inc, Arlington, Tenn), material for treatment of these bone lesions is advantageous. Calcium sulfate has a long-standing history in the treatment of bone voids secondary to trauma, infection, or neoplastic processes. Minimally-Invasive Injectable Graft injectable calcium sulfate graft may be injected into surgically created osseous defects or bone defects secondary to traumatic injury. After hardening, the paste acts as a temporary intraosseous support through which internal fixation may be placed. Fifteen patients with benign space-occupying lesions were treated with curettage and grafting with an injectable, surgical-grade, calcium sulfate bone graft substitute. Bone healing was assessed postoperatively and defined as resorption of the bone graft substitute material and replacement with new bone formation. The grafted defects in 14 of the 15 patients showed complete incorporation of the graft material at an average of 8 weeks. One patient required incision and drainage for a deep infection 5 weeks postoperatively. The average follow-up was 6 months (range: 3-15 months), and Musculoskeletal Tumor Society functional outcome scores averaged 83%. This injectable, minimally invasive graft material is easy to use, provides a temporary structural support when grafting bone defects, and results in predictable rapid bone healing. PMID- 14763546 TI - Use of percutaneous intramedullary decompression and medical-grade calcium sulfate pellets for treatment of unicameral bone cysts of the calcaneus in children. PMID- 14763547 TI - Functional outcomes of bone graft substitutes for benign bone tumors. AB - Benign bone tumors are frequently treated surgically by intralesional excision with grafting. Due to the significant morbidity associated with autogenous graft, there is interest in bone graft substitutes. Ninety-eight consecutive patients with benign bone tumors treated by intralesional excision with bone graft substitutes were retrospectively reviewed. Functional scores using the International Symposium of Limb Salvage (ISOLS) system were obtained at most recent follow-up. The mean functional score was 28.3 out of 30 (94%). Three patients suffered fractures that ultimately healed with management. There were no local recurrences. Based on results of this study, bone graft substitutes are a reasonable alternative to autogenous grafting with a successful functional outcome. PMID- 14763548 TI - Effective management of major lower extremity wounds using an acellular regenerative tissue matrix: a pilot study. AB - Wound healing is a significant problem in orthopedics. Graftjacket tissue matrix (Wright Medical Technology, Inc, Arlington, Tenn), a novel acellular regenerative tissue matrix, has been designed to aid wound closure. A prospective, randomized study was initiated to determine the efficacy of this tissue product in wound repair compared with conventional treatment. Lower extremity wounds are refractile to healing in patients with diabetes mellitus. Therefore, researchers used diabetic foot ulcers to evaluate the efficacy of GraftJacket tissue matrix in wound repair. Only a single administration of the tissue matrix was required. After 1 month of treatment, preliminary results demonstrate that this novel tissue matrix promotes faster healing at a statistically significant rate over conventional treatment. Because wounds in this series of patients are deep and circulation around the wound is poor, the preliminary results suggest that this tissue matrix will be applicable to other types of orthopedic wounds. PMID- 14763549 TI - GraftJacket augmentation of chronic Achilles tendon ruptures. PMID- 14763550 TI - An injectable calcium sulfate-based bone graft putty using hydroxypropylmethylcellulose as the plasticizer. AB - The addition of a plasticizer to synthetic bone graft substitutes can improve handling characteristics, injectability, and the ability to uniformly fill defects. Restoration of large medullary bone defects using an injectable calcium sulfate-based putty using hydroxypropylmethylcellose as the plasticizer was compared to conventional calcium sulfate paste in a canine model. Beginning 2 weeks following implantation, serial clinical and specimen radiographs demonstrated a similar progressive resorption of the implanted materials and replacement with new bone for both the putty and paste forms of calcium sulfate. The area fraction of new bone and remaining implant material in bone defects treated with the putty were not significantly different from defects treated with conventional calcium sulfate paste after 13 and 26 weeks. In addition to its handling characteristics, the putty was biocompatible and as effective as conventional calcium sulfate paste in achieving substantial bony restoration of a large, critical-size bone defect. PMID- 14763551 TI - Measurement of bone morphogenetic proteins and other growth factors in demineralized bone matrix. AB - Osteoinductivity of demineralized bone matrix has been attributed to bone morphogenetic proteins (BMP). Other growth factors, including insulin-like growth factor-1 (IGF-1) and transforming growth factor-beta1 (TGF-beta1), have also been detected in demineralized bone matrix. Success of bone graft substitutes containing demineralized bone matrix has been assumed to be closely associated with osteoinductivity of the demineralized bone matrix. Because of differences in bone characteristics between donors and tissue banks, confirmation and measurement of osteoinductivity may play a crucial role in predicting the success of the bone graft substitute. In the current studies, BMP-2, BMP-4, TGF-beta1, and IGF-1 were measured in demineralized bone matrix. A strong association was noted between BMP-2 and TGF-beta1 levels. A strong association was also found between BMP-2 and new bone formation in an ectopic nude rat model. PMID- 14763552 TI - Cellular interactions and bone healing responses to a novel porous tricalcium phosphate bone graft material. AB - The use of a porous tricalcium phosphate bone void filler (Cellplex TCP, Wright Medical Technology, Inc, Arlington, Tenn) as an alternative to autograft in bone grafting was studied in benchtop, in vitro cell culture, and in vivo preclinical studies. The experimental design included material property quantification, scaffold seeding with mesenchymal stem cells, and implantation in a rabbit segmental defect model. Measured material properties denoted appropriate composition, porosity, and strengths as compared to the literature. Fluid uptake studies and mesenchymal stem cell affinity revealed the scaffold's capabilities as a suitable host for osteoprogenitor cells. In a 1-cm rabbit diaphyseal segmental defect stabilized with an external fixator, tricalcium phosphate was compared to intact autograft, crushed autograft, and open defects. Torsional strengths and stiffnesses of tricalcium phosphate-treated tibia were greater than or equivalent to both intact and crushed autograft controls. Tricalcium phosphate pores exhibited complete bony infiltration histologically. Collectively, the tricalcium phosphate material properties, cell seeding capabilities, and in vivo biological responses give evidence of this implant's functionality as a potential alternative for autograft. PMID- 14763553 TI - EI-2128-1, a novel interleukin-1beta converting enzyme inhibitor produced by Penicillium sp. E-2128. AB - EI-2128-1, a novel interleukin-1beta converting enzyme (ICE) inhibitor, was isolated from the culture broths of Penicillium sp. E-2128. EI-2128-1 selectively inhibited human recombinant ICE activity with IC50 value of 0.59 microM, without inhibiting elastase and cathepsin B. EI-2128-1 also inhibited mature interleukin 1beta secretion from THP-1 cells induced by LPS with IC50 value of 0.28 microM. PMID- 14763554 TI - Oximidine III, a new antitumor antibiotic against transformed cells from Pseudomonas sp. I. Taxonomy, fermentation, isolation, physico-chemical properties and biological activity. AB - Our screening for antitumor antibiotics against transformed cells resulted in the isolation of a new active metabolite, oximidine III, from Pseudomonas sp. QN05727. This substance selectively inhibited the growth of rat 3Y1 fibroblasts transformed with various oncogenes. In ras- or src-transformed cells, oximidine III arrested the cell cycle at G1 phase and increased the expression of p21WAF1. PMID- 14763555 TI - Oximidine III, a new antitumor antibiotic against transformed cells from Pseudomonas sp. II. Structure elucidation. AB - The structure of oximidine III, a new antitumor antibiotic against transformed cells from Pseudomonas sp. QN05727, was determined to be a benzolactone enamide containing an O-methyloxime moiety as shown in Fig. 1 by NMR spectral analysis including a variety of two-dimensional techniques. PMID- 14763556 TI - R176502, a new bafilolide metabolite with potent antiproliferative activity from a novel Micromonospora species. AB - During the course of a screening program intended to identify new antiproliferative agents, a new bafilolide metabolite was discovered. R176502 (1) was isolated from the liquid fermentation cultures of a novel Micromonospora species found in African river bottom sediment. It was purified from ethyl acetate extracts using a series of countercurrent chromatographic steps. The structure was determined using 1- and 2-D NMR experiments. Three previously described bafilomycins (bafilomycins A1 (2), B1 (3), and B2 (4)) were also isolated (from other microbial strains). R176502 exhibited potency for inhibition of tumor cell proliferation in the nM range of concentrations. PMID- 14763557 TI - New quinones and hydroquinones from Malbranchea cinnamomea HKI 286 and HKI 296 and interaction with Tax/CREB expression system in yeast. AB - In addition to malbranicin (1) and dihydromalbranicin (5), new substituted quinones 2, 3, 6 and hydroquinone 4 were isolated from the culture brothes of two strains of Malbranchea cinnamomea. The chemical constitutions of new metabolites 2, 3, 4 and 6 were elucidated by optical spectroscopy, mass spectrometry and 1D/2D NMR spectroscopy. 2 (7-methoxymalbranicin) at a concentration of 42 microM inhibited by 67% Tax/CREB-mediated expression of beta-galactosidase in a recombinant strain of Saccharomyces cerevisiae. PMID- 14763558 TI - Acremonidins, new polyketide-derived antibiotics produced by Acremonium sp., LL Cyan 416. AB - Acremonidins A to approximately E (1 to approximately 5) were produced by fermentation of Acremonium sp., LL-Cyan 416, in heterogeneous phases. The structures of these compounds, containing a bridging keto group, were determined by spectroscopic analysis. Acremonidins A and B showed moderate activity against Gram-positive bacteria, including the methicillin-resistant staphylococci and vancomycin-resistant enterococci. Selective acylations of acremonidin B afforded ester derivatives 6 to approximately 9 that exhibited improved antibacterial activity. PMID- 14763559 TI - Bhimamycin A to approximately E and bhimanone: isolation, structure elucidation and biological activity of novel quinone antibiotics from a terrestrial Streptomycete. AB - From the ethyl acetate extract of a terrestrial Streptomycete isolate, five new quinone antibiotics, bhimamycin A (2a), B (2b), C (3c), D (5a), E (7) and the new tetralone bhimanone (8) were isolated together with the known microbial products chrysophanol (1a), aloesaponarin II (1b), 3,8-dihydroxy-1-methylanthraquinone-2 carboxylic acid (1c), adenosine, 2'-deoxyadenosine, phenylacetamide, and 2-(p hydroxyphenyl)ethanol. The structures of these natural products were deduced from the spectral data and confirmed by comparison with related compounds from the literature and by synthesis. PMID- 14763560 TI - Himalomycin A and B: isolation and structure elucidation of new fridamycin type antibiotics from a marine Streptomyces isolate. AB - In our screening of marine Streptomycetes for bioactive compounds, in addition to the known metabolites rabelomycin (1), fridamycin D (2b), N-benzylacetamide and N (2'-phenylethyl)acetamide, two new anthracycline antibiotics designated as himalomycin A (2c) and B (2d) were isolated from the culture broth of the marine Streptomyces sp. isolate B6921. The structure of the new antibiotics was determined by comparison of the NMR data with those of fridamycin D (2b) and by detailed interpretation of mass, 1D and 2D NMR spectra. PMID- 14763561 TI - pTOYAMAcos, pTYM18, and pTYM19, actinomycete-Escherichia coli integrating vectors for heterologous gene expression. AB - A novel shuttle integration cosmid vector (pTOYAMAcos), based on pKU402, and shuttle integration vectors (pTYM18 and pTYM19) were constructed for the cloning of actinomycete DNA and its heterologous expression. These vectors contain oriT of an IncP transmissible plasmid in order to transfer genes by conjugation from Escherichia coli to actinomycetes, and they also contain int derived from actinophage phiC31 in order to integrate site-specifically into the chromosomal DNA. pTOYAMAcos contains the lambdacos site to promote packaging of vectors containing 35 to approximately 45-kb DNA fragments into lambda particles. pTYM18 and pTYM19 contain kanamaycin and thiostrepton resistance genes, respectively, and have multiple cloning sites including EcoRI and HindIII sites, which are available for blue/white screening in E. coli. To demonstrate the utility of these vectors, we expressed the entire gene cluster for rebeccamycin biosynthesis from Lechevalieria aerocolonigenes using pTOYAMAcos and detected rebeccamycin production in transformed S. lividans. In addition, we demonstrated the utility of pTYM 19 in a gene-disruption complementation test. L. aerocolonigenes deltarebC strain, which is defective in rebeccamycin production because of a rebC deletion, was restored to rebeccamycin production by complemention by rebC cloned in pTYM 19. PMID- 14763562 TI - A relationship between the mevalonate pathway and isoprenoid production in actinomycetes. AB - Most Streptomyces strains are equipped with only the 2-C-methyl-D-erythritol 4 phosphate (MEP) pathway for the formation of isopentenyl diphosphate. In addition to this pathway, some Strepromyces strains have the mevalonate pathway to produce terpenoid antibiotics. We have previously shown that a gene cluster for biosynthesis of terpentecin, a diterpene antibiotic, was located in adjacent the mevalonate pathway gene cluster. In this study, a mevalonate pathway gene cluster was cloned from Actinoplanes sp. strain A40644, an isoprenoid antibiotic BE-40644 producer, to examine whether the mevalonate pathway genes and isoprenoid biosynthetic genes are clustered in genomic DNA. By sequencing flanking regions a probable BE-40644 biosynthetic gene cluster was found in the downstream region of the mevalonate pathway gene cluster. Heterologous expression of a 9-kb fragment confirmed that a set of the BE-40644 biosynthetic genes was involved in the fragment. This result suggested that the presence of the mevalonate pathway might be a good landmark to detect the production of isoprenoid compounds by actinomycetes. PMID- 14763564 TI - A new bicyclic guanidine alkaloid, Sch 575948, from a marine sponge, Ptilocaulis spiculifer. PMID- 14763563 TI - Inhibitory activity of diacylglycerol acyltransferase by cochlioquinones A and A1. PMID- 14763565 TI - Reconstruction of the cervical trachea. 1964. PMID- 14763566 TI - Suspension laryngoscopy revisited. AB - Every critical advancement in direct laryngoscopic surgical technique has enhanced its precision. Among the most notable was Killian's seminal description of suspension laryngoscopy 90 years ago, which allowed for bimanual direct laryngoscopic surgery. Because of the technical difficulties encountered while performing suspension laryngoscopy, Brunings and Seiffert designed fulcrum laryngoscope holder-stabilizers for spatula laryngoscopes from Killian's original instrument design. Their devices, which were easier to use and better tolerated by patients, were supported from the laryngeal cartilage framework or chest wall. Laryngoscope holder-stabilizers were retrofitted to tubular laryngoscope specula in the 1940s and 1950s, whereupon they became very popular. Suspension laryngoscopy should have become more common subsequent to the introduction of general endotracheal anesthesia with paralysis in the 1960s. However, laryngoscope holder-stabilizers were entrenched as the device preferred by most, and they remain so today. This entrenchment occurred despite the fact that suspension laryngoscopy allows for positioning a larger examining speculum, which in turn allows for enhanced exposure and endolaryngeal procedural precision. The applied vector forces on the mandible, maxilla, oral cavity, pharynx, and larynx associated with suspension laryngoscopy are preferable to those associated with holder-stabilizers. A prospective assessment of 120 cases revealed effective use of suspension laryngoscopy in all. We believe that only a minority of surgeons has actually seen true suspension laryngoscopy and that its merits are worthy of reexamination. PMID- 14763567 TI - Markers of wound healing in vocal fold secretions from patients with laryngeal pathology. AB - This study seeks to determine the ability of enzyme-linked immunosorbent assays of vocal fold secretions to detect and describe the inflammatory response in the vocal folds. Vocal fold and palatal secretions were collected during operation from patients with a range of vocal fold disorders and from control patients. The secretions were subjected to assays for interleukin-1beta, prostaglandin E2, and transforming growth factor beta. The results indicate a differential expression of mediators associated with the wound healing cascade in the vocal folds. The prostaglandin E2 levels clearly differentiated vocal fold secretions associated with laryngeal disease versus control sites. Furthermore, the interleukin-1beta concentrations were significantly elevated in subjects with epithelial lesions of the vocal folds as opposed to lesions of the lamina propria. Although still in its infancy, such analysis may ultimately hold scientific and clinical utility in the study and management of patients with vocal fold disease. PMID- 14763568 TI - Endoscopic management of anterior skull base encephaloceles. AB - Encephaloceles are relatively rare phenomena produced by the protrusion of brain and dura through an anterior skull base defect. Although they can occur as congenital defects, encephaloceles can also present after trauma. The diagnosis is usually made with nasal endoscopy and imaging studies. This report reviews our recent experience repairing 5 encephaloceles in 4 patients. The diagnostic approach and the technical aspects of surgical management are discussed. Although encephaloceles are a rarity, this diagnosis should be considered as part of the differential diagnosis in evaluating a patient with a unilateral polypoid nasal mass, particularly in the setting of recurrent meningitis or cerebrospinal fluid rhinorrhea. PMID- 14763569 TI - Tissue maturation during the growth of juvenile nasopharyngeal angiofibroma. AB - Juvenile nasopharyngeal angiofibroma involves a specific age group and changes from vascular tissue to fibrous tissue. The goal of this study was to investigate the correlation among tumor extension, the histologic characteristics of the tumor, and the patient's age. We performed a prospective study of 43 male patients with untreated juvenile nasopharyngeal angiofibroma (average age, 15.42 years). We correlated the patient's age with the extension of the tumor (analyzing the computed tomographic scan) and the immunohistologic characteristics of the tumor (morphological and morphometric analysis of the central portion of the tumor, the sphenopalatine foramen region). We observed that the larger the tumor, the smaller the number of vessels and cells, but the larger the fibrous component and the higher the grade of maturation of the tissue. The patient's age was not correlated with tumor extension or the histologic characteristics of the tumor. However, the grade of maturation of the tissue in the region of the sphenopalatine foramen was higher in larger tumors. PMID- 14763570 TI - Lymphocele after neck dissection. AB - The current report documents a case of lymphocele after neck dissection and reviews the management and treatment options. PMID- 14763571 TI - Resolution of hoarseness after endovascular repair of thoracic aortic aneurysm: a case of Ortner's syndrome. AB - We report the case of a 75-year-old man with a 6-month history of hoarseness due to a left recurrent laryngeal nerve palsy. Investigations revealed a thoracic aortic aneurysm compressing the left recurrent nerve; thus, the diagnosis of Ortner's syndrome, ie, cardiovocal syndrome, could be established. The aortic aneurysm was repaired by implantation of an endovascular stent graft under local anesthesia. The patient was discharged 5 days later, and at the 1-year follow-up visit the hoarseness had resolved completely. This case demonstrates for the first time the reversal of Ortner's syndrome after endoluminal aneurysm repair. PMID- 14763572 TI - Cricopharyngeal myotomy in a patient with situs inversus totalis. AB - Otolaryngologists must remain vigilant in understanding the complex anatomy of the head and neck and must always be cognizant of deviations from normal anatomy. This text discusses an interesting case that should remind surgeons of the variable course that the recurrent laryngeal nerve takes on either side of the neck and the special consideration that must be taken in performing surgery near the nerve. PMID- 14763573 TI - Factors associated with postoperative delirium after major head and neck surgery. AB - Postoperative delirium (POD) is an acute change in cognitive status characterized by fluctuating consciousness and is associated with high incidences of morbidity, high complication rates, and long hospitalizations. This study was performed to determine the incidence of POD and the perioperative risk factors in order to predict which patients have an increased risk and thus to prevent POD after major head and neck surgery. The authors retrospectively evaluated 341 patients who underwent laryngectomy or the Commando (combined operation of mouth, mandible, and neck dissection) procedure at Pusan National University Hospital from January 1986 through July 2001. Postoperative delirium developed in 13.8% of the patients who underwent laryngectomy (42 of 304) and 13.5% of the patients who underwent the Commando procedure (5 of 37). A multivariate analysis showed that older age, hypertension, low postoperative O2 saturation, and decreased postoperative hemoglobin levels were risk factors for POD (p < .05). Postoperative delirium is preventable, and its incidence can be decreased by predicting these risk factors during the preoperative and postoperative periods. PMID- 14763574 TI - Sialolithiasis of an accessory parotid gland. AB - We report a case of sialolithiasis of an accessory parotid gland in the cheek demonstrated by computed tomography and sialography. The accessory parotid gland was located anterolateral to the masseter muscle and was isolated from the main parotid gland. The calculus developed from this accessory parotid gland, and the main parotid gland was free of sialolithiasis and inflammation. To our knowledge, this is the first report concerning sialolithiasis in an accessory parotid gland. The calculus was removed without facial nerve injury or salivary fistula via a peroral approach. PMID- 14763575 TI - Small cell undifferentiated carcinoma of the submandibular gland with neuroendocrine features. AB - This article reports the clinical, histopathologic, and immunohistochemical findings in a case of small cell undifferentiated carcinoma of the submandibular gland. The tumor was composed of anaplastic cells slightly larger than lymphocytes without ductal differentiation. On immunohistochemical analysis, the tumor contained cells that reacted positively with antibodies to cytokeratin, neuron-specific enolase, synaptophysin, and chromogranin. The present case supports the hypothesis that small cell undifferentiated carcinomas of the salivary glands arise from presumed multipotential ductal stem cells. When this tumor entity is located on the salivary glands, it appears to behave less aggressively than when it is a primary tumor of the bronchial tree. PMID- 14763577 TI - Indications for mastoidectomy in acute mastoiditis in children. AB - The objective of this study was to identify clinical features of acute mastoiditis in children that are indicative of the need for mastoidectomy. We performed a retrospective chart review of 40 children (20 male, 20 female) between 2 months and 12 years 9 months of age with a diagnosis of acute mastoiditis who were managed in our institution between July 1998 and June 2002. All patients received intravenous antibiotics; this was the only treatment in 14 patients (35%). Tympanostomy tubes were inserted in 22 patients, together with postauricular needle aspiration in 12 (30%), and incision and drainage of subperiosteal abscess in 10 (25%). Mastoidectomy was performed in 4 cases (10%), and cholesteatoma was found in 3. One other child was subsequently found to have cholesteatoma. We conclude that children who present with acute mastoiditis should undergo mastoidectomy if cholesteatoma is clinically suspected, or if extratemporal suppurative complications have occurred. PMID- 14763576 TI - Comparison of human, canine, and ovine laryngeal dimensions. AB - Geometric measurement of the laryngeal skeleton is a fundamental step in laryngeal studies, especially in biomechanical modeling. Traditionally, canine larynges have been used as models of the human larynx because of their similarity in size and gross structure, but the search continues for an alternative model because of the diminishing availability of the canine species for research in the United States. In this report, a revised method for defining and labeling laryngeal framework parameters is proposed. Ovine laryngeal cartilages were measured, and the measurements were compared to those of human and canine cartilages previously reported in the literature. The ovine cricoid, thyroid, and arytenoid cartilages were significantly different from the human and canine cartilages. Also, the lack of a definite border between the true and false vocal folds revealed that the ovine model may not always be suitable for a direct comparison to the human larynx in phonation. PMID- 14763578 TI - Sudden sensorineural hearing loss after rapid reduction of blood pressure in malignant hypertension. AB - Although circulatory impairment of the cochlea would be intuitively a possible cause of sudden sensorineural hearing loss (SSHL), definite proof is lacking. A 46-year-old man developed bilateral SSHL immediately after rapid reduction of the blood pressure in malignant hypertension. Vertigo and left-sided hearing impairment and tinnitus resolved spontaneously a few hours after onset. The right sided hearing loss and tinnitus persisted, and the hearing loss improved only 17 dB after 3 months of treatment. This case may provide an in vivo human model of SSHL caused by ischemia. Bilateral involvement is possible in SSHL if the circulatory disturbance is systemic. In addition, a poorer prognosis is anticipated on the side with the more severe initial attack. PMID- 14763580 TI - Chondroid chordoma at the jugular foramen causing retrolabyrinthine lesions in both the cochlear and vestibular branches of the eighth cranial nerve. PMID- 14763579 TI - Tolerability of N-chlorotaurine in the guinea pig middle ear: a pilot study using an improved application system. AB - The tissue tolerance of N-chlorotaurine (NCT), a mild endogenous antimicrobial oxidant, has been investigated by application to the guinea pig middle ear. The animals were implanted with a novel cannula system that allows chronic external drug delivery to the round window niche. In the first part of the study, 3 animals each received 100 microL of 0.1% NCT (5.5 mmol/L) and 1% NCT, respectively, in aqueous solution twice daily for 8 days. In the second part, NCT was dissolved in phosphate-buffered saline solution to 300 milliosmolar (isotonic), and 27 microL was injected in 3 additional animals twice daily for 7 days. The guinea pigs injected with 100 microL of NCT developed immediate dizziness and nystagmus and did not thrive. Other reactions included mucosal thickening in the middle ear, rupture of the tympanic membrane, and blood and gelatinous material in the cochlea accompanied by hair cell loss and a 10- to 90 dB elevation of the hearing threshold as determined by auditory brain stem responses. The effects seemed to be dose-dependent, but the rate of variability was high across animals. In contrast, the guinea pigs treated with 27 microL of isotonic NCT showed no signs of discomfort, no or only moderate thickening of the middle ear mucosa, no shift of the hearing threshold, and no hair cell loss. Positive control animals injected with 10% neomycin sulfate developed extensive hair cell loss. Provided that the membranes of the inner ear are intact and that low single-dose volumes are used to avoid increased middle ear pressure, isotonic NCT seems to be well tolerated in the tympanic cavity. The new drug delivery system proved to be advantageous for ototoxicity studies. PMID- 14763582 TI - January blahs and gray days. PMID- 14763583 TI - Working together to ensure quality patient care. PMID- 14763584 TI - Assertiveness training to prevent verbal abuse in the OR. AB - THE HIGH INCIDENCE OF VERBAL ABUSE directed toward perioperative nurses by surgeons has been the subject of recent research studies. REPEATED INCIDENTS of verbal abuse in the OR contribute to increased incidence of errors, low morale, and high turnover among nursing staff members. ASSERTIVENESS TRAINING that focuses on conflict resolution and communication skills is an effective method of coping with verbal abuse. EACH MEMBER is a vital part of the perioperative team. Any disruption to the team, including verbal abuse, can compromise patient safety. Collaboration among team members is critical to ensure safe patient outcomes. PMID- 14763585 TI - Safe use of lasers in the operating room-what perioperative nurses should know. AB - SAFETY HAZARDS are inherent in laser use, but they can be eliminated or significantly reduced with adherence to proper procedures. THIS ARTICLE reviews beam-related and nonbeam-related safety hazards, including eye injuries, fire and thermal injuries, smoke plume, and electrical hazards, as well as safety measures to avoid these hazards. EDUCATION IN LASER SCIENCE and safety is key to recognizing potential hazards and ensuring that safety parameters are followed. PMID- 14763586 TI - Using activity-based costing in surgery. AB - ACTIVITY-BASED COSTING is an accounting technique that allows organizations to determine actual costs associated with their services based on the resources they consume. THIS TECHNIQUE can be used in a variety of ways, including targeting high-cost activities, forecasting financial baselines, and supporting resource allocation. FOUR STEPS should be followed when applying activity-based costing to surgical procedures. THIS ARTICLE explores how Froedtert Memorial Lutheran Hospital, Milwaukee, used activity-based costing. PMID- 14763587 TI - Nursing reflections from journaling during a perioperative internship. AB - AN IMPORTANT CONCERN in nursing practice and education is the difficulties nurses experience as they transition into a new clinical area. THIS STUDY compared the reflective journals of 26 experienced and inexperienced nurses participating in a nine-week perioperative internship. THE STUDY examined self-regulated learning strategies used to enhance metacognitive critical thinking abilities. PMID- 14763588 TI - It was huge! Nursing students' first experience at AORN Congress. AB - AN EXPERIENTIAL KNOWLEDGE of mentoring through nursing students' perspectives may enhance AORN's ability to recruit students to perioperative nursing and aid future planning for student involvement in the Association. IN 2003, four first year nursing students attended the AORN Congress in Chicago with their nursing instructor and mentor. The students' experiences were captured using a thematic analysis to analyze their journals. THE FIVE COMMON THEMES identified were "it was huge," "exhibits," "student program," "exploring the city," and "suggestions for future planning." PMID- 14763589 TI - Learning from stories--a pathway to patient safety. PMID- 14763590 TI - Conflict of interest. PMID- 14763591 TI - Achilles tendon rupture: an alternative treatment. AB - We have found percutaneous repair of the Achilles tendon to be a straightforward operation that can be performed under local anesthesia on an outpatient basis. Our experience leaves us with no doubt that the results in terms of wound complications and cosmesis are superior to those obtained with open operative treatment. Repeat rupture rates are comparable to the rates seen with open procedures, and patient satisfaction is excellent. Although this technique does not replace the more traditional open operative repair, it certainly has become an acceptable treatment option, and it is gaining more popularity among orthopedic surgeons. We recommend this technique without reservation. PMID- 14763592 TI - Infections of the spine: what the orthopedist needs to know. AB - Infections of the spine usually present with back pain in the absence of constitutional symptoms. Diagnosis requires a high index of suspicion and appropriate radiological and laboratory studies. Most infections can be managed conservatively with organism-specific antibiotics and bracing with good success. Surgery may be indicated to isolate the offending organism and to manage neurological and structural complications. PMID- 14763593 TI - Occult spinal cord abnormalities in children referred for orthopedic complaints. AB - This study reviews spinal cord abnormalities found in children initially referred for an orthopedic problem. Over a 5-year period in an academic pediatric orthopedic referral clinic, 167 children aged 3 months to 18 years (average, 9.4 years) underwent spine magnetic resonance imaging (MRI) scans and had records available for review. The patients were divided into 7 major groups based on the primary indication for the MRI. The frequency of spinal cord pathology was as follows: 3 of 35 patients with atypical idiopathic scoliosis, 1 of 19 with neuromuscular scoliosis, 6 of 18 with congenital scoliosis, 1 of 50 with unexplained back pain, 3 of 17 with gait abnormality, 5 of 14 with limb pain or weakness, and 4 of 8 with rigid or recurrent foot deformity. Spine MRI was not very helpful in evaluating children who had some degree of back pain without neurological signs or symptoms. However, the spine MRI was helpful in evaluating children with atypical idiopathic scoliosis or congenital scoliosis, gait abnormality, limb pain or weakness, or rigid or recurrent foot deformities. Given the high frequency of occult spinal cord abnormality in children with severe foot deformity, the use of screening spine MRI may be especially useful in this group. PMID- 14763594 TI - Pseudarthrosis after lumbar spine fusion: nonoperative salvage with pulsed electromagnetic fields. AB - We studied 100 patients in whom symptomatic pseudarthrosis had been established at more than 9 months after lumbar spine fusion. All patients were treated with a pulsed electromagnetic field device worn consistently 2 hours a day for at least 90 days. Solid fusion was achieved in 67% of patients. Effectiveness was not statistically significantly different for patients with risk factors such as smoking, use of allograft, absence of fixation, or multilevel fusions. Treatment was equally effective for posterolateral fusions (66%) as with interbody fusions (69%). For patients with symptomatic pseudarthrosis after lumbar spine fusion, pulsed electromagnetic field stimulation is an effective nonoperative salvage approach to achieving fusion. PMID- 14763595 TI - Clinical presentation and radiographic findings of distal biceps tendon degeneration: a potentially forgotten cause of proximal radial forearm pain. AB - For patients who present insidiously with proximal-radial forearm pain, the differential diagnosis should include distal biceps tendon degeneration. If radial tunnel and pronator syndromes, lateral epicondylitis, and radiocapitellar arthritis are eliminated as potential causes, tenderness over the proximal radius, supination weakness, and characteristic radiographic or magnetic resonance imaging findings may facilitate correct diagnosis of distal biceps tendon degeneration. In this article, we describe the clinical presentation and radiographic findings for 5 patients who presented with distal biceps degeneration. PMID- 14763596 TI - Double-layered patella: marker for multiple epiphyseal dysplasia. PMID- 14763597 TI - Nonunion of 5th metatarsal tuberosity fracture (dancer's fracture) associated with gout. PMID- 14763598 TI - Biplanar Kapandji intrafocal pinning of distal radial fractures. PMID- 14763599 TI - Minimally invasive subcutaneous fasciotomy for chronic exertional compartment syndrome of the lower extremity. AB - Chronic exertional compartment syndrome of the lower extremity can be debilitating in the active population. Open fasciotomy typically provides a cure or significant improvement in up to 90% of patients, provided that the compartments are adequately released. Cosmetic deformity from the surgery may be a major concern, especially in the young female athlete. A technique for minimally invasive subcutaneous fasciotomy is described that allows adequate compartment release and a favorable cosmetic result. PMID- 14763600 TI - Osteochondritis dissecans. PMID- 14763601 TI - The genetics of obesity in Mexican Americans: the evidence from genome scanning efforts in the San Antonio family heart study. AB - Recent estimates indicate that approximately 18% of the population in the United States can be considered obese (defined as a body mass index [BMI] > or = 30), and this rate is even higher among ethnic populations such as Mexican Americans. This figure becomes very significant given the strong evidence for obesity as a major risk factor for a variety of chronic diseases including type 2 diabetes mellitus and coronary heart disease. The search for genes involved in the expression of obesity has been one of the focal points of the San Antonio Family Heart Study (SAFHS), a large, family-based study to examine the genetics of risk for atherosclerosis in Mexican Americans. To date, our genome scanning effort has reported two quantitative trait loci (QTLs) with pronounced effects on the expression of a variety of obesity--related phenotypes (e.g., leptin levels, fat mass, and BMI) located on chromosomes 2 and 8. We are currently working to further refine these signals and to identify the genes and allelic variants involved. Here, we summarize the latest results from our ongoing efforts to identify obesity genes in the San Antonio Family Heart Study. PMID- 14763602 TI - Mitochondrial DNA variations in Russian and Belorussian populations. AB - The sequence of the first hypervariable segment (HVS-I) of mitochondrial DNA (mtDNA) was determined in 251 individuals from three eastern Slavonic populations, two Russian and one Belorussian. Within HVS-I, 78 polymorphic positions were revealed. Within-population diversity of HVS-I varies slightly among three samples; its estimates do not differ strongly from those for European populations. Haplotype diversity for three populations calculated in this study is 0.949; mean pairwise differences estimate is 3.59. To assign mtDNA sequences to major phylogenetic clusters, haplogroup-specific restriction polymorphisms were selectively typed in most samples. The haplogroup distribution in the total Eastern Slavonic sample is similar to that reported for the European sample. However, the separate consideration of three Slavonic samples reveals the complicated structure of the mitochondrial gene pool in the Eastern European area. Data of this study support the proposed model of the origin of modern Eastern Slavs, which implies the admixture of ancient Slavonic tribes with pre Slavonic populations of Eastern Europe. These data should contribute to general studies of mitochondrial DNA variations in Europe. PMID- 14763603 TI - Molecular and temporal characteristics of human retropseudogenes. AB - One of the primary forces driving genome evolution is retrotranscription. In addition to creating new genetic material from which new genes with new functions arise, retrotranscription leaves traces of its action in the form of retropseudogenes. These loci, which are intronless, retrotransposed copies of mature mRNAs from functional antecedent genes, are layered throughout genomes as a molecular fossil record of genome evolution. A survey of 138 functional source genes in the human genome has revealed more than three hundred retropseudogenes. Analysis of the characteristics of the source genes shows that, on average, their size, G/C content, and expression patterns fit the canonical features of source genes reported elsewhere. Details of insertion site duplications for these loci are consistent with a model of retropseudogene formation involving endogenous retrotranscription and enzymatic mobilization and retroposition. Retrotranscription event age estimates reveal a pattern in which the highest densities appear after major phylogenetic events in primate history and then decline. This temporal pattern suggests that the processes forging genome evolution are most active during periods of speciation and adaptive radiation and then steadily diminish until the next burst of activity. PMID- 14763604 TI - Microsatellite diversity among three endogamous Tamil populations suggests their origin from a separate Dravidian genetic pool. AB - The genetic profiles based on 15 autosomal microsatellite markers were analyzed among three socially distinct endogamous Dravidian populations: Tanjore Kallar, Vanniyar, and Pallar of Tamil Nadu, southern India, in order to understand their origin and the extent of genetic affinity and diversity among them. All loci were highly polymorphic and followed Hardy-Weinberg expectations except for loci D13S317 in Tanjore Kallars and D7S820 in Vanniyars. The SK2 criterion test showed no evidence of association among the 15 loci in the studied populations. The extent of gene differentiation among the three populations was low (G(ST) = 0.012), suggesting proximity between them. The phylogenetic dendrogram based on allele frequencies places them in a separate cluster, away from other compared Indo-European populations. The fit of the Harpending and Ward model of regression was found to be good and consistent with the extent of endogamy followed by the respective populations. These findings support a separate origin of the Dravidians and reveal an overall genetic unity among the studied Tamil populations belonging to different strata of the social hierarchy. The extent of diversity found among them probably resulted from the strict endogamous practices that they follow. PMID- 14763605 TI - Further studies of the influence of apolipoprotein B alleles on glucose and lipid metabolism. AB - The effect of five genetic polymorphisms in the apolipoprotein B gene on parameters of lipid and glucose metabolism was assessed in 564 Danish mono- and dizygotic twins. Genotypes in apolipoprotein B T71I (ApaLI RFLP), A591V (AluI RFLP), L2712P (MvaI RFLP), R3611Q (MspI RFLP), and E4154K (EcoRI RFLP) were established using polymerase chain reaction and restriction enzyme digests. The effect of genotypes on lipid levels and on glucose, insulin, and HOMA (i.e., calculated parameters of beta-cell function and insulin resistance) was assessed by multivariate analyses of variance correcting for the effect of gender, age, glucose tolerance status, and body mass index. The effect of genotype on the risk of having impaired glucose metabolism was calculated by logistic regression analysis. Finally, linkage between allele sharing and physiological parameters was calculated by the new Haseman-Elston method. The allele frequencies of all five polymorphisms were similar to those previously reported for Caucasian populations. The L2711P (MvaI RFLP) polymorphism influenced LDL-cholesterol and LDL-to-HDL measures (p = 0.04 and 0.03, respectively), while the R3611Q (MspI RFLP) polymorphism had an effect on the insulin-to-glucose ratio (p = 0.04), and E4154K (EcoRI RFLP) influenced HOMAbeta (p = 0.04). Significant interactions were observed between genotype in T71I (ApaLI RFLP), A591V (AluI RFLP), R3611Q (MspI RFLP), and E4154K (EcoRI RFLP) and glucose tolerance on lipid-related parameters (0.03 < p < 0.004), and between genotype in L2712P (MvaI RFLP) and E4154K (EcoRI RFLP) and gender on lipid and glucose-related parameters (0.02 < p < 0.003). No genotypes were significantly associated with impaired glucose tolerance measured by logistic regression. Likewise, no effect of allele sharing in the five polymorphisms was seen in the dizygotic twins. The effect of the polymorphisms on lipid and glucose parameters could be mediated through linkage to genes with known effect on glucose metabolism or through free fatty acids exerting their effect on glucose metabolism. PMID- 14763606 TI - More than a leap of faith: the impact of biological and religious correlates on reproductive behavior. AB - Using a conceptual model that integrates both social and biomedical factors of causation, this paper tries to delineate the pathways through which the reproductive characteristics of a multidenominational community are characterized. In total, 5513 historical entries from family reconstitution were available. Selection of data was guided by the inclusion of information about religious affiliation. Only married couples with children as well as single mothers with the relevant information were considered. Of these, 1855 entries were of Roman Catholic (C), 1143 of Lutheran/Protestant (L/P2), and 609 of Reformed Calvinist (R) denomination. The analysis documented differential nuptiality and fertility patterns, which at first glance may be interpreted along religious lines. However, the paper attempts to show that these various sociocultural patterns associated with religious behavior are merely proximate determinants, while the ultimate causes are biological in nature (i.e., differential parental age at marriage or birth, different parity progression regimes, differences in median interpregnancy interval, as well as highly variable sibship size within the denominational groups). PMID- 14763607 TI - Fertility and mortality differentials among the population groups of the Himalayas. AB - Selection potential based on differential fertility and mortality has been computed for 24 Himalayan populations classified into four groups: Himalayan populations with Asian affinities (HPA), Brahmans, Rajputs, and Scheduled Castes (Shilpkars). Irrespective of the methodology followed, the total index of selection was found to be highest (0.794) in Barbatiyas (HPA) and lowest (0.428) in Rajputs. The relative contribution of fertility component (If) to the index of total selection (It) is higher than the corresponding mortality component (Im) in all 24 groups. An analysis of correlation and regression on different components of the indices of selection performed among these 24 Himalayan populations revealed that the contributions of If in determining It are stronger than Im. Further, both If and Im are strongly associated with It and account for 76% and 67% of total variability in It, respectively. Examination of the relationship of the selection potential with the differential altitude and social categories showed a decrease in the index from high altitude to low altitude. Similarly, an inverse relationship was found between various indices and social categories. Himalayan populations with Asian affinities (HPA) in the present study reveal higher values of selection potential. Interestingly, these populations invariably reside in high altitude areas where health and communication infrastructures are poorly developed. Thus, the differential pattern of fertility and mortality among the Himalayan populations indicates that they are passing through a stage of transition, as is evident from the decrease in various selection indices. It is also apparent that groups that are less developed socioeconomically, as well as those inhabiting high altitude regions, are lagging behind in this process. PMID- 14763608 TI - A modified locally weighted method for developing reference standards for height, weight, and body mass index of boys and girls aged 4 to 18 in Taiwan. AB - The objectives of this study are to modify the locally weighted method to obtain reference values for height, weight, and body mass indices (BMI): to describe the median growth velocity of children aged 4 to 18 in Taiwan; and to compare those values with data from other countries. We modified the LOCAL method to generate empirical percentiles and used the locally weighed regression to smooth the percentiles. We examined the smoothed percentiles against original data to ensure that smoothed percentiles were within 1% of errors. We used numerical differentiation of the 50th percentiles of height and weight to obtain the velocity of growth in height and weight, respectively. We applied our method to the data of the Nutrition and Health Survey in Taiwan, 1993-1996, which was a multistaged stratified sample. The sample included 2351 boys and 2355 girls aged 4 to 18. Our results indicate that (a) the proposed method, which utilizes the original data, guarantees the percentiles within 1% errors; (b) before puberty, the height, weight, and BMI of Taiwanese children are not much different from those of their counterparts in Western countries; and (c) after puberty and at the stage of young adulthood, the height, weight, and BMI of Taiwanese youth are below the levels of those in Western young adults. Genetic makeup and dietary patterns may contribute to these phenomena. PMID- 14763609 TI - Restriction isotyping of apolipoprotein E among populations of Punjab, northwestern India. AB - The molecular polymorphism displayed by apolipoprotein E (apoE, protein; APOE, gene) has been listed as a risk factor for susceptibility to various disorders, such as those associated with lipid metabolism and arteriosclerosis. Data from many population groups are available. The present study endeavors to add to the world population database for alleles encountered at this locus. One hundred sixty-five individuals representing four castes and a mixed group from Punjab, a state in northwestern India, were analyzed for APOE isotyping. Intercaste group comparisons of allele frequencies revealed statistically insignificant differences, pointing to homogeneity at this locus among Punjabi caste groups, which can be considered as one Punjabi population. A further comparison of this Punjabi sample with other populations of the world revealed the Punjabi population to be closer to some European populations than to either African or Asian populations, a pointer to the ethnic origins of the Punjabi population. PMID- 14763610 TI - Notice of plagiarism. PMID- 14763611 TI - Apology from N. Samani and M. Pasandi. PMID- 14763612 TI - Field evaluation of emergency well disinfection for contamination events. PMID- 14763613 TI - The journey from safe yield to sustainability. AB - Safe-yield concepts historically focused attention on the economic and legal aspects of ground water development. Sustainability concerns have brought environmental aspects more to the forefront and have resulted in a more integrated outlook. Water resources sustainability is not a purely scientific concept, but rather a perspective that can frame scientific analysis. The evolving concept of sustainability presents a challenge to hydrologists to translate complex, and sometimes vague, socioeconomic and political questions into technical questions that can be quantified systematically. Hydrologists can contribute to sustainable water resources management by presenting the longer term implications of ground water development as an integral part of their analyses. PMID- 14763615 TI - Influence of calibration methodology on ground water flow predictions. AB - We constructed a numerical model of transient ground water flow and solute transport for a portion of the Biscayne Aquifer in Florida, and calibrated the model with three different combinations of data from a 193-day period: head (h) data alone, data on h and ground water discharge to a canal (q), and data on h, q, and ground water chloride concentration (C). We used each of the three calibrated models to predict h and q during a 182-day test period separate from the calibration period. All three calibrated models predicted h equally well during the test period (r = 0.95, where r = 1 indicates perfect agreement between measured and simulated values), though the model calibrated on h alone had significantly different parameter values than the other two models. Predictions of q during the test period depended on calibration methodology; models calibrated with multiple targets simulated q more accurately than the model calibrated on h alone (r = 0.79 compared to r = 0.49). Based on the results of these simulations, we conclude: (1) Post-calibration prediction is important in assessing the value of different data types in automated calibration; (2) inverse solution uniqueness is not a requirement for accurate h predictions; (3) relatively simple models can predict with reasonable accuracy transient ground water flow in a complex aquifer, and parameters governing this prediction can be estimated by nonlinear regression methods that incorporate both h and q data; (4) addition of C data to the calibration did not improve model predictive capacity because the information in the C data was similar to that in the q data, from the perspective of model calibration (the subsurface chemical signal in question was controlled mainly by seepage of high-chloride canal water into the low-chloride ground water system). PMID- 14763614 TI - Sources of ground water salinity on islands using 18O, 2H, and 34S. AB - Stable isotopes of 18O and 2H in water, and 34S and 18O in dissolved SO4, are used to verify the interpretation of the chemical evolution and proposed sources of salinity for two islands that have undergone postglacial rebound. Results for delta18O and delta34S in dissolved SO4 on the Gulf Islands, southwest British Columbia, Canada, suggest a three-component mixing between (1) atmospheric SO4 derived largely from recharge of meteoric origin, (2) modern marine SO4 associated with either modern-day salt water intrusion or Pleistocene age sea water, and (3) terrestrial SO4. The age of the marine SO4 is uncertain based on the geochemistry and SO4 isotopes alone. Two options for mixing of saline ground waters are proposed--either between current-day marine SO4 and atmospheric SO4, or between older (Pleistocene age) marine SO4 and atmospheric SO4, delta18O and delta2H compositions are relatively consistent between both islands, with a few samples showing evidence of mixing with water that is a hybrid mixture of Fraser River water and ocean water. The isotopic composition of this hybrid water is approximately delta18O = 10 per thousand. delta18O and delta2H values for many saline ground waters plot close to the global meteoric water line, which is distinctly different from the local meteoric water line. This suggests a meteoric origin for ground waters that is different from the current isotopic composition of meteoric waters. It is proposed these waters may be late Pleistocene in age and were recharged when the island was submerged below sea level and prior to rebound at the end of the last glaciation. PMID- 14763616 TI - Multiple-aquifer characterization from single borehole extensometer records. AB - Measurement and analysis of aquifer-system compaction have been used to characterize aquifer and confining unit properties when other techniques such as flow modeling have been ineffective at adequately quantifying storage properties or matching historical water levels in environments experiencing land subsidence. In the southeastern coastal plain of Virginia, high-sensitivity borehole pipe extensometers were used to measure 24.2 mm of total compaction at Franklin from 1979 through 1995 (1.5 mm/year) and 50.2 mm of total compaction at Suffolk from 1982 through 1995 (3.7 mm/year). Analysis of the extensometer data reveals that the small rates of aquifer-system compaction appear to be correlated with withdrawals of water from confined aquifers. One-dimensional vertical compaction modeling indicates measured compaction is the result of nonrecoverable hydrodynamic consolidation of the fine-grained confining units and interbeds, as well as recoverable compaction and expansion of coarse-grained aquifer units. The calibrated modeling results indicate that nonrecoverable specific storage values decrease with depth and range from 1.5 x 10(-5)/m for aquifer units to 1.5 x 10( 4)/m for confining units and interbeds. The aquifer and Potomac system recoverable specific storage values were all estimated to be 4.5 x 10(-6)/m, while the confining units and interbeds had values of 6.0 x 10(-6)/m. The calibrated vertical hydraulic conductivity values of the confining units and interbeds ranged from 6.6 x 10(-4) m/year to 2.0 x 10(-3) m/year. These parameter values will be useful in future management and modeling of ground water in the Virginia Coastal Plain. PMID- 14763617 TI - Geophysical monitoring and evaluation of coastal plain aquifers. AB - We use time domain electromagnetic (TDEM) soundings to monitor ground water conditions beneath the coastal plain in eastern North Carolina. The TDEM method measures the earth's response to an induced electromagnetic field. The resulting signal is converted, through a complex inversion process, to apparent resistivity values, which can be directly correlated to borehole resistivity logs. TDEM soundings are used to map the interface between fresh and salt water within coastal aquifers, and estimate depth to basement when siting new monitoring wells. Focused TDEM surveys have identified areas of salt water encroachment caused by high volumes of discharge from local supply wells. Electromagnetic sounding, when used in tandem with the state's network of monitoring wells, is an accurate and inexpensive tool for evaluating fresh water/salt water relationships on both local and regional scales within coastal plain aquifers. PMID- 14763618 TI - Use of an electromagnetic seepage meter to investigate temporal variability in lake seepage. AB - A commercially available electromagnetic flowmeter is attached to a seepage cylinder to create an electromagnetic seepage meter (ESM) for automating measurement of fluxes across the sediment/water interface between ground water and surface water. The ESM is evaluated through its application at two lakes in New England, one where water seeps into the lake and one where water seeps out of the lake. The electromagnetic flowmeter replaces the seepage-meter bag and provides a continuous series of measurements from which temporal seepage processes can be investigated. It provides flow measurements over a range of three orders of magnitude, and contains no protruding components or moving parts. The ESM was used to evaluate duration of seepage disturbance following meter installation and indicated natural seepage rates resumed approximately one hour following meter insertion in a sandy lakebed. Lakebed seepage also varied considerably in response to lakebed disturbances, near-shore waves, and rainfalls, indicating hydrologic processes are occurring in shallow lakebed settings at time scales that have largely gone unobserved. PMID- 14763619 TI - Modeling the buildup of iron oxide encrustations in wells. AB - Ferric iron encrustations are a common problem that seriously affects the performance of wells and drains. Their formation is induced by the mixing of reduced ground water containing ferrous iron with oxic shallow ground water and exposure to air. The process of ferrous iron oxidation is a kinetically controlled reaction. The reaction rate has a quadratic dependency on pH. The precipitating oxides have an autocatalytic effect that further enhances reaction progress. This paper describes the application of kinetic models to the problem of encrustation formation. Influences of pH, residence time, and autocatalysis were modeled. The overall influence of the autocatalytic reaction path is particularly strong when initial amounts of iron oxides are present. Autocatalysis provides a good explanation on the development of well yield commonly measured in the field. Encrustation precipitation is slow at first, but speeds up after a sufficient amount of iron oxide has built up. An incomplete removal of iron oxide encrustations during rehabilitation leads to a renewed increase in catalytic efficiency and encrustation precipitation. PMID- 14763620 TI - Use of temperature profiles and stable isotopes to trace flow lines: Nagaoka area, Japan. AB - In this study, we use borehole temperature data and stable isotopes to delineate the flow system and estimate the effect of urbanization in the Nagaoka area of Japan. Temperature profiles were measured four times in observation wells during the period 2000-2001 and compared with those measured in the same wells during the period 1977-1983 (Taniguchi 1986). Water was sampled in both observation and pumping wells during the same period. The temporal and spatial variability in temperature indicate clearly the effect of urban warming and heavy pumping on the ground water system. Urban warming caused higher temperatures recently as compared to the older values, and pumping caused induced recharge from the river to the ground water. The stable isotope data show the ground water flow system is divided into shallow, intermediate, and deep systems, and that land use and infiltration rate are affecting the shallow flow system. PMID- 14763621 TI - Semianalytical solutions for stream depletion in partially penetrating streams. AB - In the analysis of streamflow depletion, the Hunt (1999) solution has an important advantage because it considers a partially penetrating stream. By extending the Hunt drawdown solution, this paper presents semianalytical solutions for gaining streams that evaluate the induced stream infiltration and base flow reduction separately. Simulation results show that for a given deltah (the initial hydraulic head difference between stream and aquifer beneath the channel), the base flow reduction is in direct proportion to the product of streambed leakage (lambda) and the distance between pumping well and stream (L), and the induced stream infiltration is in inverse proportion to lambdaL. Deltah has a significant effect on the ratio of stream infiltration to base flow reduction. The results from the semianalytical solutions agree well with those from MODFLOW simulations. The semianalytical solutions are useful in the verification of numerical simulations and in the analysis of stream-aquifer interactions where water quantity or quality is concerned. PMID- 14763622 TI - Quantifying ground water recharge at multiple scales using PRMS and GIS. AB - Management of ground water resources requires a method to calculate demonstrably accurate recharge rates at local to regional scales using readily available information bases. Many methods are available to calculate recharge, but most are unable to satisfy all these conditions. A distributed parameter model is shown to meet the stated needs. Such models are input intensive, however, so a procedure to define most inputs from GIS and hydrogeological sources is presented. It simplifies the PRMS calibration observed streamflow hydrographs by reducing degrees of freedom from dozens to four. For seven watersheds (60 to 500 km2), the GIS-aided calibrations have average errors of 5% on recharge and 2% on total streamflow, verifying the accuracy of the process. Recharge is also calculated for 63 local-scale subwatersheds (average size 37 km2). For the study area, calculated recharges average 11 cm/yr. Soil and rock conductivity, porosity, and depth to the water table are shown to be the physical properties which dominate the spatial variability of recharge. The model has been extended to uncalibrated watersheds where GIS and climatic information are known. It reproduces total annual discharge and recharge to within 9% and 10%, respectively, indicating the process can also be used to calculate recharge in ungauged watersheds. It has not been tested outside the study area, however. PMID- 14763623 TI - Small-scale electrical resistivity tomography of wet fractured rocks. AB - This paper describes a series of experiments that tested the ability of the electrical resistivity tomography (ERT) method to locate correctly wet and dry fractures in a meso-scale model. The goal was to develop a method of monitoring the flow of water through a fractured rock matrix. The model was a four by six array of limestone blocks equipped with 28 stainless steel electrodes. Dry fractures were created by placing pieces of vinyl between one or more blocks. Wet fractures were created by injecting tap water into a joint between blocks. In electrical terms, the dry fractures are resistive and the wet fractures are conductive. The quantities measured by the ERT system are current and voltage around the outside edge of the model. The raw ERT data were translated to resistivity values inside the model using a three-dimensional Occam's inversion routine. This routine was one of the key components of ERT being tested. The model presented several challenges. First, the resistivity of both the blocks and the joints was highly variable. Second, the resistive targets introduced extreme changes the software could not precisely quantify. Third, the abrupt changes inherent in a fracture system were contrary to the smoothly varying changes expected by the Occam's inversion routine. Fourth, the response of the conductive fractures was small compared to the background variability. In general, ERT was able to locate correctly resistive fractures. Problems occurred, however, when the resistive fracture was near the edges of the model or when multiple fractures were close together. In particular, ERT tended to position the fracture closer to the model center than its true location. Conductive fractures yielded much smaller responses than the resistive case. A difference-inversion method was able to correctly locate these targets. PMID- 14763624 TI - Predictability of surface mine spoil hydrologic properties in the Appalachian Plateau. AB - Prevention of acid mine drainage at surface coal mines in the Appalachian region relies to an extent on minimizing ground water contact with acid-forming materials, and maximizing ground water contact with alkalinity-yielding materials. Acid-forming materials are often selectively handled to minimize or prevent contact with ground water. Controlling ground water contact with acidic or alkaline materials depends on forecasting the level and range of fluctuation of the postmining water table within the mine backfill. Physical measurements and aquifer testing of more than 120 wells from 18 reclaimed mines in Kentucky, Ohio, Pennsylvania, and West Virginia have led to improved forecasting of the postmining ground water system. Factors that influence the ground water regime include spoil lithology and particle size, age of reclamation, spoil thickness, distance from the final highwall, and pit floor dip angle and direction. Spoil hydraulic conductivity (K) exhibits a 95% confidence interval range of six orders of magnitude about a mean K of 1.7 x 10(-5) m/sec. Spoil aquifer saturated thickness is related to the overall thickness of the spoil, the lithology of the spoil, dip of the pit floor, and distance to the highwall. Saturated spoil thickness has a 95% confidence interval of 2.2 to 3.6 m about the mean of 2.9 m. The predicted saturated zone averages 19% of the total spoil thickness. PMID- 14763625 TI - Transient analysis of the source of water to wells: Cape Cod, Massachusetts. AB - A transient flow modeling analysis for potential public-supply wells on western Cape Cod, Massachusetts, demonstrates the difference between transient and steady state recharge areas can have important implications for wellhead protection. An example of a single pumping well illustrates that commonly used steady-state time related capture areas do not represent the recharge area and travel times of water being pumped from the well until sufficient time has elapsed for steady state flow conditions to be established. Until that time, transient recharge areas are needed to account for the portion of water discharging from the well that entered the aquifer before pumping started. An example of two pumping wells demonstrates the same area at the water table cannot supply water to more than one well under steady-state flow conditions. Transient recharge areas to multiple wells can overlap, however, until steady-state flow conditions are established. The same area can, therefore, be a source of water to more than one well during early pumping times, and the water pumped from a given well may derive from source areas, including contaminated areas, that do not lie within the well's steady-state recharge area. PMID- 14763626 TI - "Methods to determine storativity of infinite confined aquifers from a recovery test," by Djaouida Chenaf and Robert P. Chapuis, July-August 2002 issue, v. 40, no. 4: 385-389. PMID- 14763627 TI - "Implications of observed and simulated ambient flow in monitoring wells," by Alper Elci, Fred J. Molz III, and W. R. Waldrop, November-December 2001 issue, v. 39, no. 6: 853-862. PMID- 14763628 TI - "Analytical model for aquifer response incorporating distributed stream leakage," by Garey A. Fox, Paul DuChateau, and Deana S. Durnford, July-August 2002 issue, v. 40, no. 4: 378-384. PMID- 14763629 TI - Personal reflections on Saratoga Springs, New York: hydrogeological and horse racing 'hot spot' of the East. PMID- 14763630 TI - The proof is in the process. PMID- 14763631 TI - A vision for our future. PMID- 14763632 TI - Documenting outcomes of occupational therapy: the center for outcomes research and education. AB - This article presents two themes related to the development and implementation of sound outcomes research that emerged from a 3-year dialogue among fellows of the University of Illinois Center for Outcomes Research and Education (CORE). These themes are: that outcomes research needs to be an interrelated multi-method system of investigation, and it should be developed in collaboration with its key stakeholders (consumers and practitioners). Examples from the literature and from a cluster of papers published in this focus issue are used to exemplify the themes. PMID- 14763633 TI - Randomized controlled trials to investigate occupational therapy research questions. AB - The randomized controlled trial (RCT) has become the most widely accepted research design for testing the efficacy of health care interventions. The purposes of this paper are to (a) introduce the essential elements of RCTs, and (b) discuss some of the special problems faced by occupational therapists in conducting and interpreting RCTs. The CONSORT (Consolidated Standards of Reporting Trials) statement is recommended as an introduction to the components of RCT quality. Problems pertinent to the study of the efficacy of occupational therapy and related interventions include the importance of: theory, background, and rationale; treatment fidelity; theory-based outcomes; management of non masked (non-blinded) interventionists and participants; and multiplicity of statistical analyses. This paper can help practitioners judge the quality of an RCT, and it can help the profession work toward the development of a cadre of qualified researchers who can adapt the well-established methods of RCTs to the study of occupation-based interventions. PMID- 14763634 TI - Quality of life and symptom severity for individuals with chronic fatigue syndrome: findings from a randomized clinical trial. AB - OBJECTIVE: Chronic fatigue syndrome is a profoundly disabling condition characterized by severe, unrelenting fatigue and a number of other physical and cognitive symptoms. Currently, there is no cure or widely accepted treatment for chronic fatigue syndrome, and few rehabilitation programs exist to address quality of life issues in chronic fatigue syndrome. In the present randomized clinical trial, the effects of an integrative, consumer-driven rehabilitation program on quality of life and symptom severity for individuals with chronic fatigue syndrome were examined. METHOD: Forty-seven participants were randomly assigned to either an immediate program group (n = 23) or a delayed program control group (n = 24) and assessed with the Chronic Fatigue Syndrome Symptom Rating Scale and the Quality of Life Index before the program, after program participants completed the group phase, and after program participants completed the one-on-one phase. It was hypothesized that the program would lead to improvements in quality of life and an overall reduction in symptom severity. RESULTS: Linear growth models were estimated comparing program and control conditions over time using random-effects regression analyses. Significant condition by time interactions were observed for the main outcomes of symptom severity and overall quality of life. Effect sizes for these interactions involving symptom severity (Cohen's d = 0.71) and overall quality of life (Cohen's d = .66) were moderate. CONCLUSIONS: Findings indicate that consumer driven programs such as this one can have a positive impact on symptom severity and quality of life over time for individuals with chronic fatigue syndrome. PMID- 14763635 TI - Impact of pediatric rehabilitation services on children's functional outcomes. AB - OBJECTIVES: Relatively little is known about the treatment effectiveness and functional outcomes of pediatric rehabilitation therapies. This study was conducted to gain knowledge of the type and quantity of inpatient rehabilitation services provided to children who received acute inpatient rehabilitation and compare functional gains by age and diagnosis. METHOD: A retrospective cohort design was used. Records of rehabilitation therapies and functional assessments of 814 pediatric patients who received inpatient rehabilitation during 1996, 1997, and 1998 were collected. The admission and discharge item ratings of the WeeFIM instrument were first transformed into interval-level measures of self care, mobility, and cognition. Parametric analyses were used to compare functional gains across impairment groups and to examine the relationship between amount of treatment and functional gains. RESULTS: Occupational therapy and physical therapy were the primary rehabilitation services received by patients across impairment groups (98% and 99%, respectively). A large proportion of children with traumatic brain injuries also received speech therapy (97%) and psychology services (60%). Across domains (self-care, mobility, cognition), the largest gains were made by children who were older than 7 years and had traumatic injuries. Functional gains were significantly related to the amount of discipline specific treatment received, after controlling for age, impairment, and functional status at admission. CONCLUSION: Rehabilitation therapy provision in pediatric inpatient rehabilitation varies greatly depending on children's age and the nature of the impairment. Systematic reporting of type and quantity of rehabilitation therapies along with functional assessments before and after hospitalization would allow researchers to track functional changes and study the determinants of functional improvement. PMID- 14763636 TI - Concerns about the future among older adults with multiple sclerosis. AB - Multiple sclerosis (MS) is a chronic, progressive neurological disease that causes demyelination of the central nervous system. Typically diagnosed in adulthood, it does not significantly reduce life expectancy. The goal of this exploratory study was to describe the health-related concerns and service needs of 27 older adults with MS, ages 55 to 81 years. Through in-depth interviews using a phenomenological approach, fear of the future was found to be a predominant concern among the participants. Within this fear, participants expressed particular concerns about experiencing further losses of mobility and independence, becoming a burden on caregivers, and having to move to a nursing home. The findings raise three major challenges for occupational therapists that include: (1) developing or modifying interventions that can enable older adults with MS to gain a sense of control over their future, (2) working with families affected by MS together with other disciplines such as psychology and social work, and (3) advocating for more and better community support options for persons with MS. PMID- 14763637 TI - Outcomes of a vocational program for persons with AIDS. AB - Advances in medical treatment combined with changes in the demographics of persons who are becoming infected with autoimmune deficiency syndrome (AIDS) have transformed this illness from a rapidly progressing to a chronically disabling condition in a short period of time. This paper describes the development, implementation, and outcomes of a program of vocational services for persons with AIDS. This program was studied using a single group design, in which participatory action research strategies were used to investigate and improve the program as it unfolded. In addition to examining the overall outcomes of services, the study aimed to discover which components were most helpful to participants and which participants were most likely to benefit from the program. Of 129 participants of who initially enrolled, 39 dropped out before finishing the program. Sixty of the 90 participants who completed the program achieved employment, returned to school, or began a volunteer position or internship. Consequently, the overall success rate was 46.5% and the success rate for program completers was 66.7%. The occupational narrative, which participants told in their initial assessment interview, was closely associated with both program completion and successful outcomes. This association adds support to the importance of narrative for understanding participants and predicting future behavior, as well as for the therapy process. PMID- 14763638 TI - Developing and evaluating community-based services through participatory action research: two case examples. AB - Occupational therapy has a strong history of embracing concepts of client empowerment. However there is limited literature in the field on how to achieve empowerment, or on how to extend empowerment to the level of the community and social groups and services within it. This article discusses how concepts and strategies of participatory action research, an extension of empowerment theory, can be used to inform service development and evaluation in occupational therapy. The participatory action research approach is illustrated using two case examples of participatory action research programs for persons with chronic fatigue syndrome and individuals with autoimmune deficiency syndrome (AIDS). A critical analysis of the application of this approach to research and practice is provided. Finally, the paper identifies key principles of participatory action research that can be used to guide occupational therapy services and empower both individuals and communities. PMID- 14763639 TI - Mothering a child with hidden impairments. AB - When a mother has a child with a chronic impairment, the occupational demands of mothering extend to address the specialized needs of that child. This research explores how the type of hidden impairment in a child influences family routines and occupations. This qualitative study consisted of interviews with 22 mothers of children with attention deficit hyperactivity disorder (ADHD), a behavioral disorder, and 22 mothers of children with cystic fibrosis, a physical disorder. Open-ended questions were used to explore family demands, resources, time use, routines, concerns, and support. The transcripts of these interviews were analyzed in terms of consistency with existing literature on parenting the child with hidden disability, and for emergent themes. In this analysis the experiences of mothers of children with cystic fibrosis were consistent with research findings on other chronic conditions, with these mothers reporting that family provides them with extensive physical and emotional support. Although mothers of children with cystic fibrosis reported a persistent emotional sorrow, they felt they were able to "normalize" their daily routines. In contrast, mothers of children with ADHD reported little family support, high perception of child related demands, and less confidence in their success in mothering these children. In describing their daily routines, these mothers often stated that there was no such thing as a "normal" day. They felt constantly "on alert" and did not feel that they had "normal" routines. Based on this study, mothers of children with ADHD felt distress because their child did not easily conform to social standards, and were likely to express exhaustion in their role as "mother." The pattern of responses offered by these participants differs significantly from that of the participants whose children have cystic fibrosis, and from the usual pattern of coping with chronic childhood disability documented in the literature. PMID- 14763640 TI - The test-retest reliability of the child-initiated pretend play assessment. AB - OBJECTIVE: The purpose of this study is to establish the test-retest reliability of the Child-Initiated Pretend Play Assessment (ChIPPA) (Stagnitti, 2002a; Stagnitti, Unsworth, & Rodger, 2000). METHOD: The first author rated 38 preschool children ages 4 and 5 years (4 with developmental delay and 34 typically developing) on the ChIPPA. The ChIPPA employs conventional play materials and unstructured play materials to assess three qualities of a child's play ability: elaborateness of play action, ability to substitute objects during play, and the child's need to imitate the modelled actions of the examiner. The ChIPPA was administered twice, at a 2-week interval, to each participant. RESULTS: Test retest intraclass correlation coefficients (ICCs) (Type 2,1) calculated for each of the three elaborate play measures ranged from .73 to .84. A test-retest ICC of .56 was obtained for object substitution with unstructured play materials. The test-retest ICC obtained for the combined score for unstructured and conventional play materials was .57. Percentage agreement figures ranging from 63.2% to 84.2% were obtained on test-retest of the object substitution with conventional toys and imitated actions measures. There was no significant difference between test and retest scores for these measures based on a Wilcoxon Matched Pairs Signed Ranks Test (Wilcoxon Sign Test). CONCLUSION: Elaborate play scores, object substitution with conventional toys score, and imitation scores on the ChIPPA showed stability over time. Object substitution scores using unstructured materials were the least stable play measures and appeared to be related to the child's play themes. Since play is the primary occupation of children, it is essential that therapists have a reliable measure of play behavior. The test retest reliability results from the ChIPPA provide evidence that this assessment produces a stable measure of play behavior that can then guide therapists when planning intervention strategies for children. PMID- 14763641 TI - Convergent and predictive validity of three scales related to falls in the elderly. AB - The purpose of this study was to determine if the Activities-specific Balance Confidence (ABC) Scale, Falls Efficacy Scale (FES), and Survey of Activities and Fear of Falling in the Elderly (SAFE) assessments measured fear of falling in the same manner (convergent validity) and to determine if they predicted those individuals who, based upon a previous history of falls, limitation of activity, and not leaving home, had an increased potential for falling (predictive validity). One hundred and eighteen individuals, 60 years of age and older, completed each of the assessments. They self-reported activity restriction, fall history, and the number of times they left the home each week. The convergent validity of each assessment was established by correlating each assessment tool with each of the others. Findings indicated the ABC and FES were highly correlated with each other, indicating they measured similar constructs, and both were moderately correlated with the SAFE, suggesting these assessments measured different constructs. The predictive validity of each instrument in relation to the frequency of falls, limitation of activity, and frequency of leaving the home revealed no individual tool could accurately predict any of these characteristics of the sample. As a result, no one test by itself was able to identify individuals who may be at risk and a candidate for an intervention program. PMID- 14763642 TI - Interrater reliability and discriminant validity of the deductive reasoning test. AB - OBJECTIVE: The purpose of this study was to determine the interrater reliability and discriminant validity of Toglia's Deductive Reasoning test for individuals with brain injuries. METHOD: Forty-two individuals with brain injuries and 51 participants without disabilities ranging in age from 18 to 84 years were given the Deductive Reasoning test three consecutive times. A between groups repeated measure design was used to examine differences between the two groups in performance of the Deductive Reasoning test across the three trials. RESULTS: An interclass correlation coefficient (ICC) was statistically significant, indicating good interrater reliability for participants without disabilities. A repeated measures analysis of variance (ANOVA) comparing performance across trials and between groups revealed a significant group main effect, F(1,91) = 52.68, p < .001, thus, participants with brain injuries performed significantly lower on the test than participants without disabilities. This analysis also revealed a significant increase in all participants' scores across trials, F(1.63, 148.67) = 35.094, p < .001. CONCLUSION: Findings confirm acceptable interrater reliability of the Deductive Reasoning test. Further, comparison of the performance of the two groups supports the discriminant validity of the Deductive Reasoning test for individuals with brain injuries. Thus, the Deductive Reasoning test was found to be an appropriate, valid, and reliable assessment tool for occupational therapists to assess deductive reasoning skills in individuals with brain injuries. Additional research is needed to continue to investigate the psychometric value of the Deductive Reasoning test so that clinicians and researchers can use it appropriately. PMID- 14763643 TI - The bridge from ethics to public policy: implications for occupational therapy practitioners. PMID- 14763644 TI - Understanding and implementing positive reinforcement as an intervention strategy for children with disabilities. PMID- 14763645 TI - Value based management. PMID- 14763646 TI - Caring work, personal obligation and collective responsibility. AB - Studies of workers in health care and the care of older people disclose tensions that emerge partly from their conflicting obligations. They incur some obligations from the personal relationships they have with clients, but these can be at odds with organizational demands and resource constraints. One implication is the need for policies to recognize the importance of allowing workers some discretion in decison making. Another implication may be that sometimes care workers can meet their obligations to clients only by taking collective action. PMID- 14763647 TI - Expanding nurses' participation in ethics: an empirical examination of ethical activism and ethical assertiveness. AB - This research project investigated the extent to which nurses engage in two important kinds of ethical behaviours: ethical activism (where they try to make hospitals more receptive to nurses' participation in ethics deliberations) and ethical assertiveness (where they participate in ethics deliberations even when not formally invited). This research probed not only the extent to which nurses engage in these ethical behaviours but also whether this is influenced by professional, training and organizational factors. A random sample of 165 nurses from three major hospitals in Los Angeles provided the data. Regression analyses indicate that both ethical activism and ethical assertiveness are strongly influenced by nurses' perceptions of the receptivity of hospitals to their inclusion in ethics deliberations. In addition, nurses' education in ethics is a significant predictor of ethical activism. The findings have important implications for the content of nurses' ethics training as well as for expanding the boundaries of nurses' participation in ethics deliberations. The authors define ethics deliberations as specific meetings of a number of people to discuss an ethical issue, such as one regarding the care of a patient. PMID- 14763648 TI - An intercultural nursing perspective on autonomy. AB - This article is based on an empirical study regarding ethical challenges in intercultural nursing. The focus is on autonomy and disclosure. Autonomy is a human capacity that has become an important ethical principle in nursing. Although the relationship between autonomy and patients' possibly harmful choices is discussed, the focus is on 'forced' autonomy. Nurses seem to equate respect with autonomy; it seems to be hard to cope with the fact that there are patients who voluntarily undergo treatment but who actively participate neither in the treatment offered nor in making choices regarding that treatment. Nurses' demand for patients to be autonomous may in some cases jeopardize the respect, integrity and human worth that the ethical principle of autonomy is meant to ensure. Even though respect for a person's autonomy is also respect for the person, one's respect for the person in question should not depend on his or her capacity or aptitude to act autonomously. Is autonomy necessarily a universal ethical principle? This article negates this question and, through the issues of culture, individualism versus collectivism, first- and second-order autonomy, communication and the use of family interpreters, and respect, an attempt is made to explain why. PMID- 14763649 TI - Involvement and (potential) influence of care providers in the enlistment phase of the informed consent process: the case of AIDS clinical trials. AB - This article draws on ethnographic field data collected during an investigation of the informed consent process and AIDS clinical trials. It describes the involvement of care providers (physicians, nurse practitioners, physician assistants) during the enlistment, or recruitment, phase of the informed consent process. It shows that sometimes care providers are involved in the receipt, evaluation and distribution of information on clinical trials through their interactions with research professionals and patients. It suggests that the involvement of care providers has the potential to influence the informed consent process. Some of the ethical and practice considerations of this are discussed. PMID- 14763650 TI - Ethical moments in practice: the nursing 'how are you?' revisited. AB - In seeking for an understanding of ethical practices in health care situations, our challenge is always both to recognize and respond to the call of individuals in need. In attuning ourselves to the call of the vulnerable other an ethical moment arises. Asking 'how are you?' in health care practice is our very first possibility to learn how a particular person finds herself or himself in this particular situation. Here, 'how are you?' shows itself as an ethical question that opens up a relational space that calls forth a response. It is a way to understand the situated moments in which we are already that enables us to act respectfully. Our ethical frameworks assist us in trying to decide what is the right thing to do given a set of circumstances. Yet there is a prior step that already calls us to ethical attention; this is when we ask 'how are you?', which transforms a seemingly small interaction into an ethical moment. 'How are you?' is a question that turns us back to who we are as health care professionals and calls us to be more deeply attentive to the moment. When we sincerely ask 'how are you?' we enact our ethical commitments to one another. PMID- 14763651 TI - Caring about--caring for: moral obligations and work responsibilities in intensive care nursing. AB - The aim of this study was to analyse experiences of moral concerns in intensive care nursing. The theoretical perspective of the study is based on relational ethics, also referred to as ethics of care. The participants were 36 intensive care nurses from 10 general, neonatal and thoracic intensive care units. The structural characteristics of the units were similar: a high working pace, advanced technology, budget restrictions, recent reorganization, and shortage of experienced nurses. The data consisted of the participants' examples of ethical situations they had experienced in their intensive care unit. A qualitative content analysis identified five themes: believing in a good death; knowing the course of events; feelings of distress; reasoning about physicians' 'doings' and tensions in expressing moral awareness. A main theme was formulated as caring about--caring for: moral obligations and work responsibilities. Moral obligations and work responsibilities are assumed to be complementary dimensions in nursing, yet they were found not to be in balance for intensive care nurses. In conclusion there is a need to support nurses in difficult intensive care situations, for example, by mentoring, as a step towards developing moral action knowledge in the context of intensive care nursing. PMID- 14763652 TI - The concept of futility in health care decision making. AB - Life saving or life sustaining treatment may not be instigated in the clinical setting when such treatment is deemed to be futile and therefore not in the patient's best interests. The concept of futility, however, is related to many assumptions about quality and quantity of life, and may be relied upon in a manner that is ethically unjustifiable. It is argued that the concept of futility will remain of limited practical use in making decisions based on the best interests principle because it places such high demands on the individual responsible for decision making. This article provides a critical analysis of futility (in the context of the best interests decision-making principle), and proposes an ethically defensible notion of futility. PMID- 14763653 TI - Terrorism--the scientific response. March 2003. BMA Memorandum of Evidence to the Science and Technology Committee (HC): Inquiry into The Scientific Response to Terrorism. PMID- 14763654 TI - Interview with Fekadu Aga. PMID- 14763655 TI - Masked hypertension--a new entity and challenge for clinical hypertension research and management. PMID- 14763656 TI - Hypertension treatment and stroke prevention. PMID- 14763657 TI - Central obesity and hypertensive renal disease: association between higher levels of BMI, circulating transforming growth factor beta1 and urinary albumin excretion. AB - OBJECTIVE: In this study, the relationship between circulating transforming growth factor beta1 (TGFbeta1) and urinary albumin excretion (UAE) has been investigated in non-obese and central obese hypertensive patients. DESIGN AND PATIENTS: Fifty-eight consecutive hypertensive outpatients both lean and with central obesity were enrolled and divided in three groups, according to their body mass index (BMI) values. Group A: 16 lean hypertensives (men with BMI < 25 kg/m2 and women with BMI < 24.7 kg/m2); Group B: 16 overweight hypertensives (men with BMI > or = 25 kg/m2 and < 30 kg/m2 and women with BMI > 24.7 kg/m2 and < 27.3 kg/m2); Group C: 26 obese hypertensives (men with BMI > or = 30 kg/m2 and women with BMI > or = 27.3 kg/m2). MEASURES: In all patients, UAE, by immunonephelometric assay, circulating TGFbeta1 by a solid-phase specific sandwich enzyme-linked immunosorbent assay (ELISA) technique, blood urea nitrogen (BUN) and creatinine, by routine laboratory methods, were determined. In addition, left ventricular telediastolic internal diameter (LVIDd), interventricular septum diastolic (IVSTd), posterior wall thickness (PWT), total and normalized to height2.7 left ventricular mass (LVM, LVM/h2.7), relative wall thickness (RWT) and left ventricular ejection fraction (EF) by M-B Mode echocardiography were calculated. RESULTS: Overweight and obese hypertensives had significantly (p < 0.05) higher BMI, waist-hip ratio (WHR), UAE and TGFbeta1 than lean hypertensives. Obese hypertensives had significantly (p < 0.05) higher total and indexed LVM values than lean hypertensives. Obese hypertensives had significantly (p < 0.05) higher BMI, UAE and TGFbeta1 than overweight hypertensives. In all subjects, TGFbeta1 correlated directly with BMI (r = 0.52; p < 0.0001), WHR (r = 0.48; p < 0.003), MBP (r = 0.31; p < 0.02) and UAE (r = 0.57; p < 0.0001). Multiple regression analysis indicated that BMI, MBP and UAE were able to explain the 47.9% TGFbeta1 variability (r = 0.69; p < 0.0001), and that TGFbeta1 was the best predictor of UAE changes (r = 0.60; p < 0.0001). CONCLUSION: Our data suggest that TGFbeta1 levels are positively associated with BMI, MBP and UAE in hypertensive subjects. This also indicates that TGFbeta1 overproduction might be considered a pathophysiology mechanism of progressive renal function impairment in obese hypertensives. PMID- 14763658 TI - Hyperuricemia, low urine urate excretion and target organ damage in arterial hypertension. AB - BACKGROUND: Hyperuricemia can be the consequence of an increased urate production, a decreased renal excretion, or both. An increased prevalence of hyperuricemia has been described in essential hypertensive patients partly due to a decreased renal urinary urate excretion (UUE). Hyperuricemia has been shown to be associated with an increased risk of cardiovascular disease in hypertensive patients in some but not in all epidemiological studies in which this relationship has been investigated. OBJECTIVE: To assess the influence of low UUE in the association between serum urate, renal function and hypertension severity. PATIENTS AND METHODS: This cross-sectional study was carried out in a sample of 677 male hypertensive patients, aged 35-60 years, with essential arterial hypertension consecutively attended in a hospital hypertension unit. The presence of hypertension-related organ damage at diagnosis was classified according to classical WHO criteria as grade 1, 2 or 3. Urate underexcretion was defined as 24 h urinary urate below the product serum urate x 100. RESULTS: Mean serum urate levels were 6.4 +/- 1.6 mg/dl in the total sample. Hyperuricemia (serum urate >7 mg/dl) was present in 28.5% of patients and only 17.0% had underexcretory hyperuricemia. This subgroup of patients exhibited the higher rate of hypertension-related target organ damage (TOD). A multivariate analysis, showed that underexcretory hyperuricemia but not hyperuricemia remained an independent predictor of TOD (odds ratio 2.5. 95% CI 1.6-3.89). Serum urate correlated positively with serum creatinine in hyperuricemic patients (r = 0.50, p < 0.001), but not in patients with underexcretory hyperuricemia (r = 0.21, p = 0.18). CONCLUSIONS: Underexcretory hyperuricemia is strongly related to hypertensive organ damage and this relationship does not seem to be mediated by a decreased renal function. This aspect could underline the predictive value of hyperuricemia independently of serum creatinine. UUE could improve the clinical predictive value of hyperuricemia as a cardiovascular risk factor. PMID- 14763659 TI - Echocardiographic assessment of the different left ventricular geometric patterns in middle-aged men and women in Tallinn. AB - The aim of this study was to determine the association of left ventricular (LV) geometry with sex, age, arterial hypertension and obesity in Tallinn. In a framework of a population study for cardiovascular risk factors, echocardiography was carried out in 325 men and 398 women (69.3% of all 1043 participants aged 35 59) in 1999-2001. Left ventricular hypertrophy was defined if left ventricular mass (LVM), LVM/height and LVM/body surface area were 294 g, 163 g/m and 150 g/m2 in men, and 198 g, 121 g/m and 120 g/m2 in women, respectively. LV geometry was analysed according to four types generally recognized (with regard to relative wall thickness > 0.45). The prevalence of concentric hypertrophy was similar in men and women: 7.7% and 9.1%. The prevalence of eccentric hypertrophy was significantly higher in women than in men (33.3% vs 4.9%). Concentric remodelling was also found in women more often than in men (9.5 vs 5.5%; p < 0.05). Regardless of sex and age, concentric hypertrophy was never found in participants with blood pressure < 140/90. In hypertensives, there was a tendency for age related increase of concentric hypertrophy prevalence: the latter was higher in women than in men: 39.1% vs 25.5%; p < 0.05. In examinees with BMI < 30, this type of LV geometry was seldom found: in 3.1% of men and 5.0% of women; p < 0.05. In obese persons, it increased with age, reaching 26.5% in men and 21.2% in women (p < 0.05). The prevalence of eccentric hypertrophy in men increased with age, and with hypertension and obesity. The prevalence of concentric remodelling in men was not related to BMI; it was significantly more often found in older age groups and in hypertensives. In women, the prevalence of eccentric hypertrophy and concentric remodelling was not related to age, hypertension or obesity. PMID- 14763660 TI - Influence of different types of stressors on the waveform of the peripheral arterial pulse in humans. AB - BACKGROUND: To characterize further the different reflected waves visible at the peripheral arterial pressure waveform, we recorded changes in the pulse wave contour during different stressful interventions. METHODS: Ten young healthy volunteers underwent two protocols. At the first occasion, the digital arterial pressure waveform was continuously recorded with the Portapres device during 5 min of rest, a mental arithmetic stress test (MAT) and during the cold pressor test (COP). At a second occasion, the pulse waveform was recorded at rest and during bicycle exercise. RESULTS: The reflected pressure peak seen during systole arrived earlier and was increased in amplitude during both MAT and COP compared to rest (p < 0.05). The first reflected pressure peak during diastole arrived earlier and was reduced in amplitude compared to the systolic peak during MAT (p < 0.01), while COP induced smaller effects. Bicycle exercise at 100 and 140 heartbeats/min caused the reflected peak in systole to be undetectable in most subjects and induced a marked reduction in the amplitude of the first diastolic reflected peak (p < 0.01), and time to its appearance was markedly reduced at 140 beats/min. CONCLUSION: Several stressors accounted in daily life, such as mental stress, pain and exercise-induced rapid changes in the arterial pulse waveform. In terms of timing and amplitude, the reflected systolic pressure peak and the first diastolic pressure peak did not always change in the same direction during the interventions, implying different origins of these two reflected waves. Combining the stressors used with pulse wave analysis is a simple method for studying vascular reactivity non-invasively. PMID- 14763661 TI - Ethnic, geographic and dietary influences upon vasoactive hormones and blood pressure among Greenland Inuit and Danes. AB - AIM: To study levels of vasoactive hormones and urinary excretion of sodium and potassium between groups of Greenland Inuit and Danes, and to analyse the relationship between these hormones and 24-h blood pressure, including nightly blood pressure dips and pulse pressure. METHODS: 145 Greenlandic participants were categorized in three groups according to degree of westernization, based on dietary habits and current place of residence; 41 Danes were included as controls. Twenty-four-hour blood pressure was measured. Venous plasma concentrations of vasoactive hormones were measured. Urine was collected for 24 hours for analysis of excretion of sodium and potassium. RESULTS: The Inuit population of Greenland had a lower diastolic blood pressure, a higher pulse pressure and lower nocturnal blood pressure dip than Danes had. Angiotensin II in plasma and urine excretion of potassium were higher among Greenlanders compared with Danes, irrespective of diet and place of residence. Aldosterone and urine excretion of sodium were significantly higher among participants in Denmark compared with participants in Greenland. Brain natriuretic peptide and atrial natriuretic peptide were independently and negatively associated with diastolic blood pressure, and vasopressin was positively associated with systolic blood pressure and pulse pressure. Ethnic differences in the effect of vasoactive hormones or urinary sodium and potassium excretion could not explain the difference in blood pressure. CONCLUSION: It is suggested that a high dietary intake of potassium and low sodium intake among Greenlanders may affect blood pressure. Further attention should be drawn to the occurrence of high pulse pressure and high activity in the renin-angiotensin system in Inuit populations. PMID- 14763662 TI - Target organ damage and changes in arterial compliance in white coat hypertension. Is white coat innocent? AB - The aim of this study was to perform an extensive evaluation of target organ status, metabolic abnormalities and hemodynamic alterations in white coat hypertension (WCH). Fifty normotensive (NT), 90 WCH (ambulatory daytime blood pressure < 135/85 mmHg) and 101 hypertensive (HT) subjects underwent extensive biochemical, echocardiographic, fundoscopic examination. In a subgroup study, arterial compliance and intima-media thickness (IMT) were measured by Doppler ultrasound in left common carotid artery. WCH subjects were found to have higher body mass index (BMI) than the NTs (p = 0.042). Left ventricle mass index (LVMI) was greater in the WCHs than the NTs (p < 0.001), but significantly less than the HTs (p < 0.001). Hypertensive retinopathy was observed in the WCHs, but was less severe and rare compared to the HTs (13% vs 27%). Both WCHs and HTs had high levels of urinary albumin excretion (UAE) (p = not significant). Total cholesterol was higher in WCHs than in the NTs (p = 0.04) The distensibility coefficient (DC) of the WCHs was significantly greater than the HTs (p < 0.01), while significantly smaller than the NTs (p < 0.01). The compliance coefficient (CC) of the WCHs was significantly higher than the HTs (p < 0.01), and significantly less than the NTs (p < 0.01). The IMT in the HTs was significantly higher than the WCHs (0.81 +/- 0.05 vs 0.70 +/- 0.04 mm; p < 0.001) and the NTs (p < 0.001). The difference between the NTs and the WCHs was not significant. Our data indicate that patients with WCH represent an intermediate group between NTs and sustained HTs where target organ damage and cardiovascular risk is concerned. PMID- 14763663 TI - Exercise blood pressure is related to insulin resistance in subjects with two hypertensive parents. AB - INTRODUCTION: Insulin resistance has been implicated in the pathogenesis of essential hypertension. Elevated blood pressure during physical exercise is a more powerful predictor of future hypertension than resting blood pressure. We have therefore studied the relationship between insulin resistance and blood pressure response to exercise in strongly hypertension-prone subjects. MATERIAL: Twenty-five normotensive subjects aged 18-35 years with bi-parental hypertension, and 26 matched controls with normotensive parents. METHODS: (i) Maximal exercise tolerance test with continuous gas-exchange measurement; (ii) blood pressure at rest, during exercise and 24-h ambulatory; (iii) euglycemic hyperinsulinemic clamp. RESULTS: Diastolic blood pressure during exercise was higher in hypertensionprone subjects as compared to controls, but only in subjects with low insulin sensitivity. Resting and 24-h diastolic blood pressure were higher in hypertension-prone subjects, but independent of insulin sensitivity level. Insulin sensitivity and exercise capacity were similar in the groups. CONCLUSION: Diastolic blood pressure during exercise was higher in hypertension-prone subjects as compared to controls, but only in insulin-resistant subjects. Since elevated blood pressure during physical exertion is a predictor of future hypertension, these findings may suggest that insulin resistance is involved in the pathogenesis of essential hypertension. PMID- 14763664 TI - Comparison of the echocardiographic and pulmonary function test findings in orderly treated and untreated essential hypertensive patients. AB - OBJECTIVE: Although it has been well established that hypertension effects pulmonary functions negatively, the effect of regular antihypertensive therapy on pulmonary functions is not known. In this study, we aimed to compare the pulmonary function tests of the hypertensive patients taking regular antihypertensive therapy with those of the ones not taking any antihypertensive medicine, to document the differences in pulmonary functions of both hypertensive groups. MATERIALS AND METHODS: Patients who had received antihypertensive treatment (Group I: 29 males, 24 females, mean age 42.3 +/- 8.2 years), and untreated cases (Group II: 28 males, 22 females, mean age 43.4 +/- 6.4 years) were included in the study. Patients with a history of coronary heart disease, respiratory diseases, smokers and those who were obese were excluded from the study. RESULTS: Forced vital capacity (FVC) and forced expiratory volume at the first second (FEV1) levels were found significantly lower in Group II (p < 0.05). In echocardiographic evaluation, the mitral E/A ratio of Group II was lower than that of Group I (p < 0.05). Isovolumetric relaxation time and deceleration time was higher in Group II than in Group I (p < 0.05). There was a significant association between pulmonary function tests and impaired left ventricular diastolic parameters; especially, mitral E/A ratio was significantly associated with decreased FVC and FEV1 in Group II (respectively r = -0.695, p = 0.01 and r = -0.591, p = 0.03). CONCLUSION: FVC and FEV1 levels in untreated hypertensive cases were lower than in those of treated hypertensive cases, and this may be caused by diastolic function rather than the systolic function. In the evaluation of dyspnea in hypertensive patients, it would be useful to examine respiratory function in addition to echocardiographic investigation. PMID- 14763665 TI - Home versus Office blood pressure MEasurements: Reduction of Unnecessary treatment Study: rationale and study design of the HOMERUS trial. AB - The Home versus Office MEasurements, Reduction of Unnecessary treatment Study (HOMERUS) is a multicentre prospective study, primarily designed to examine in subjects with mild to moderate hypertension whether treatment decisions based on home blood pressure measurements can lead to reduction in the use of antihypertensive drugs and the associated costs, compared to office blood pressure measurements. After inclusion, 360 patients are randomized to two groups. In one group, antihypertensive therapy is based on blood pressure measured in the outpatient clinic: the office pressure (OP) group. In the other group, antihypertensive therapy is based on home blood pressure measurements: the self-pressure (SP) group. All readings, both in OP and in SP, are obtained with the same validated oscillometric device, the Omron 705 CP. Treatment decisions are taken by an independent physician at the coordinating centre, who is unaware whether the patient belongs to the SP or OP group. Following a standardized treatment schedule, blood pressure is targeted at 120-139 mmHg for systolic and 80-89 mmHg for diastolic pressure. Patients are followed for 1 year. At the start and at the end of the study, ambulatory blood pressure measurements are obtained as a reference. Microalbuminuria and echocardiography are assessed to evaluate the possible development of target organ damage. It is expected that, at the end of the trial, patients in both groups will have the same blood pressure, at the expense of more medication in the OP group. Therefore, a cost-minimization analysis will be performed first. If short-term effects appear not to be comparable for OP and SP, a cost-effectiveness analysis will be performed to assess the value of the SP strategy in comparison to standard practice. In addition, medication compliance is recorded within random subgroups of the SP and OP groups by means of Medication Event Monitoring System (MEMS) V TrackCaps. PMID- 14763666 TI - Accuracy of SpaceLabs 90207 is altered by venous blood redistribution. AB - Validation protocols that have been introduced for automated blood pressure (BP) measuring devices recommend procedures carried out at rest only. We aimed to determine whether venous blood redistribution affects the accuracy of the oscillometric method of BP measurement. For this purpose, we chose a popular oscillometric ambulatory BP monitor--the SpaceLabs 90207 (Osc). Lower body negative pressure (-40 mmHg) (LBNP) was used to simulate changes of body position. Fifty-one young healthy volunteers had their BP measured simultaneously by Osc and mercury sphygmomanometer (HgS) at rest (min 3 and 5), during LBNP (min 7 and 9) and after LBNP (min 11). Differences (delta, mmHg) between HgS and Osc for systolic (SBP) and diastolic BP (DBP) were calculated for every measurement minute. For SBP, deltaSBP-7 and deltaSBP-9 were significantly different from deltaSBP-5 (0.65 +/- 2.6 and 0.33 +/- 2.4 mmHg vs -0.80 +/- 2.9 mmHg, p < 0.003 and p < 0.02, respectively). deltaSBP-11 also differed significantly from deltaSBP-3 and deltaSBP-5 (1.16 +/- 2.5 mmHg vs -0.06 +/- 3.1 mmHg and -0.80 +/- 2.9, p < 0.01 and p < 0.00007, respectively). When graded according to British Hypertension Society protocol, Osc remained in the A class in every measurement minute. The accuracy of SpaceLabs 90207 is altered by venous blood redistribution. These inaccuracies may constitute an additional limitation of oscillometric ambulatory BP monitoring. PMID- 14763667 TI - Postural hypotension in hypertensive patients. AB - BACKGROUND: The association between orthostatic hypotension and supine hypertension (Hypo-Hyper) has been reported in previous studies on selected populations. The present longitudinal study aimed to characterize the haemodynamic patterns, comorbidity, prognostic implications and eventual effect of therapy in patients with the Hypo-Hyper pattern. METHODS: Ambulatory blood pressure monitoring (AMAP) and clinical evaluation were performed on 615 consecutive patients recruited from the Hypertension Clinics of five Italian Hospitals: 34 patents were identified as Hypo-Hyper, and underwent 10-month follow-up. RESULTS: The incidence of the Hypo-Hyper pattern was 5.5% in the hypertensives studied. Hypo-Hyper was more frequent in the elderly (mean age 58 years), and the affected population exhibited different kinds of underlying pathologies. Multivariate analysis showed no association between antihypertensive treatment and Hypo-Hyper pattern. Patients with AMAP features of higher blood pressure values at night-time than at daytime displayed higher rates of myocardial hypertrophy and pacemaker implantation during the follow-up. CONCLUSIONS: Although the study design did not aim to identify any pathophysiological mechanism for Hypo-Hyper pattern, these first data show that the Hypo-Hyper association is the effect of a particular subtype of hypertension, with significantly different prognostic implications. PMID- 14763668 TI - Resistant hypertension. PMID- 14763669 TI - Microalbuminuria in Type-1 diabetes mellitus. PMID- 14763670 TI - Interactions between antihypertensive agents and other drugs. PMID- 14763671 TI - Psychiatry, technology, and the corn fields of Iowa. PMID- 14763672 TI - The Internet's impact on the practice of psychiatry. AB - OBJECTIVE: In the past decade, the Internet has rapidly evolved into a mainstream communication and information resource. During this period, health care resources and databases have been developed. This paper reviews the literature on e-mail communication and on-line health care information resources to determine the impact on psychiatry. METHOD: The Medline electronic database and the search engines Altavista and Google were used to review the literature discussing on line guidelines, clinical trials, and continuing medical education (CME) resources, as well as the literature on Internet usage, e-journals, and e-mail communication. CONCLUSIONS: The Internet has accelerated psychiatrists' access to new research findings and other information pertinent to clinical practice. It has eliminated distance barriers and offered psychiatrists and the general public equal access to scientific articles, clinical trials, and guidelines. The literature shows that the general public is using the Internet to obtain information and, to a lesser extent, to communicate with health care professionals. In terms of health outcomes, the effectiveness of patient on-line access to information is still uncertain, and further research is necessary. Although physicians have embraced on-line journals, further work is required before they embrace on-line CME. The Internet has developed to a point where it is important for psychiatrists to understand the issues and pitfalls in its use, to be able to access the resources available, and to be in a position to mentor patients who want to research their condition further. PMID- 14763673 TI - Clinical and educational telepsychiatry applications: a review. AB - OBJECTIVE: Telepsychiatry in the form of videoconferencing brings enormous opportunities for clinical care, education, research, and administration. Focusing on videoconferencing, we reviewed the telepsychiatry literature and compared telepsychiatry with services delivered in person or through other technologies. METHODS: We conducted a comprehensive review of telepsychiatry literature from January 1, 1965, to July 31, 2003, using the terms telepsychiatry, telemedicine, videoconferencing, effectiveness, efficacy, access, outcomes, satisfaction, quality of care, education, empowerment, and costs. We selected studies for review if they discussed videoconferencing for clinical and educational applications. RESULTS: Telepsychiatry is successfully used for various clinical services and educational initiatives. Telepsychiatry is feasible, increases access to care, enables specialty consultation, yields positive outcomes, allows reliable evaluation, has few negative aspects in terms of communication, generally satisfies patients and providers, facilitates education, and empowers parties using it. Data are limited with regard to clinical outcomes and cost-effectiveness. CONCLUSIONS: Telepsychiatry is effective. More short- and long-term quantitative and qualitative research is warranted on clinical outcomes, predictors of satisfaction, costs, and educational outcomes. PMID- 14763674 TI - Portable computing in psychiatry. AB - Personal digital assistants (PDAs) or handheld computers have become more popular in the practice of medicine. These devices have implications for psychiatric practice, and this paper reviews various published articles describing their use. The literature demonstrates their use in document editing, patient tracking, accessing medical information and drug reference guides, prescription writing, and medical education. Security on these devices is covered, and the value and increasing implementation of PDAs in psychiatry is discussed. PMID- 14763675 TI - Assessing and monitoring antipsychotic-induced movement disorders in hospitalized patients: a cautionary study. AB - OBJECTIVE: To assess the amount of documentation and level of assessment provided by attending physicians and nursing staff in regard to extrapyramidal symptoms (EPS) experienced by hospitalized patients with varied DSM-IV diagnoses regularly treated with antipsychotic medication. METHOD: We examined the medical records of 204 hospitalized patients. All medical records were examined retrospectively from consecutive admissions beginning in January 1996. We identified demographics, length of hospitalization, diagnosis, and antipsychotic and adjunct medication. EPS were classified into dystonia, parkinsonism, akathisia, and tardive dyskinesia (TD). For each type of EPS, 2 independent raters rated the quality of assessment based on dimensions of severity, location, and laterality. RESULTS: The extent of interrater agreement was found to be 91.1%. Parkinsonism and akathisia were more frequently assessed, compared with TD and dystonia. However, the medical records examined showed generally poor assessment and documentation of EPS. The percentage of medical records with "no description" for each EPS classification was as follows: dystonia (89%), parkinsonism (71%), akathisia (67%), and TD (94%). CONCLUSIONS: The major finding of this study was a high rate of failure to document the assessment and course of EPS. This finding suggests that clinicians do not recognize the importance of documenting these significant adverse events. This shortcoming can be corrected with increased awareness of EPS and increased training in their physical examination. PMID- 14763676 TI - Sociodemographic factors associated with comorbid major depressive episodes and alcohol dependence in the general population. AB - OBJECTIVES: To estimate the 12-month prevalence of alcohol dependence (AD) among subjects with major depressive episodes (MDEs) and the 12-month prevalence of MDEs among those with AD; to investigate the associations between demographic and socioeconomic characteristics and comorbid MDE and AD, based on established theoretical models; and to compare the rates of mental health service use between groups having high and low risk for comorbid conditions. METHODS: We used data from the 1996-1997 Canadian National Population Health Survey. MDE and AD were measured using the World Health Organization's Composite International Diagnostic Interview Short Form (CIDI-SF). We calculated the 12-month prevalence of MDEs among participants with AD and of AD among those with MDEs. The associations between demographic and socioeconomic characteristics and comorbidity were investigated. RESULTS: Of participants with MDEs, 8.6% had AD; 19.6% of participants with AD reported having at least 1 MDE in the past 12 months. Being young (aged 12 to 24 years); being divorced, separated, or widowed; and having low family income level were positively associated with MDE, AD, and comorbidity. Among participants with comorbid MDE and AD, those who were aged 12 to 24 years were less likely to have used any mental health services in the past 12 months than were others. CONCLUSIONS: Young age, single marital status, and low family income may be potential risk factors for comorbid MDE and AD. Although AD is rare in the general population, public health interventions that target the groups identified as at risk may help to prevent MDE, AD, and comorbidity. PMID- 14763677 TI - Delineating the population served by a mobile crisis team: organizing diversity. AB - OBJECTIVE: While mobile crisis teams have proliferated to address gaps in service for individuals with serious mental illness, research into their effectiveness is limited. This study identifies specific cohorts of individuals served by a single mobile crisis team so that appropriate program evaluation could be designed and conducted for each unique grouping. METHOD: This retrospective study analyzes specific data from 4 sets of client records (n = 401) at 4 different time periods, using a qualitative, text-based approach. For the first 2 reviews (n = 69 and n = 40), only data related to referral source, sex, details of the reason for referral, and the mental status assessment were known to the researchers. The first 2 sets of records were used to identify the cohorts, the third set was used to confirm these cohorts, and the fourth set was used to determine the reliability of the classification tool. RESULTS: All individual cases for the mobile crisis team could be classified into one of the following categories: Symptoms Disturbing Others, Symptoms Disturbing Self, Information Seeking, For Your Information, and Other. After the third review, the cluster, Symptoms Disturbing Others, was subdivided into Symptoms Disturbing Others-General and Symptoms Disturbing Others-Suicide. CONCLUSIONS: This review was useful in identifying specific cohorts of individuals served by a single mobile crisis team. Clearly identifying who is served by a mobile crisis team is the first step in identifying practice guidelines and appropriate evaluation measures for each distinct group. PMID- 14763678 TI - Detecting women at risk for postnatal depression using the Edinburgh Postnatal Depression Scale at 2 to 3 days postpartum. AB - OBJECTIVE: This study evaluates the capacity of the Edinburgh Postnatal Depression Scale (EPDS) implemented in the first days postpartum to detect women who will suffer from postnatal depression. METHOD: A sample of 1154 women completed the EPDS at 2 to 3 days postpartum and again at 4 to 6 weeks postpartum. RESULTS: There was a highly significant positive correlation between EPDS scores on both occasions (Spearman rank correlation: r = 0.59, P < 0.0001). The cut-off scores of 10 and 11 for EPDS administered at 2 to 3 days obtained good specificity, sensitivity, and positive predictive values for the cut off scores proposed for the diagnosis of postnatal depression at 4 to 6 weeks postpartum. CONCLUSION: The EPDS completed at 2 to 3 days postpartum is a useful means of detecting women at risk of postnatal depression. PMID- 14763679 TI - Ethics in psychiatric research: study design issues. AB - OBJECTIVE: To identify and discuss ethical aspects of study design issues in psychiatric research. METHOD: We conducted a literature review and conceptual analysis of study design in psychiatric research focusing on placebo, medication tapering and withdrawal (washout), and symptom provocation (challenge) designs. RESULTS: While advances in the care and treatment of persons with psychiatric disorders are crucial to the improved well-being of this stigmatized and often forgotten population, past abuses demonstrate the importance of the thoughtful application of ethical principles in the conduct of research. Some ethical issues have particular relevance to psychiatric research arising primarily from the specific vulnerabilities of those with mental illness and the risks posed by some research methodologies. Accordingly, sensitivity is required in the design of psychiatric research. CONCLUSION: Placebo, challenge, and washout study designs can present particular risks in the population of persons with mental illness. These issues are described and suggestions offered to promote the ethical design of psychiatric research. PMID- 14763680 TI - The prevalence of psychological morbidity in West Bank Palestinian children. AB - OBJECTIVE: To determine the prevalence of psychological morbidity among Palestinian children living in the southern Bethlehem District of the West Bank during July 2000. METHODS: We undertook a descriptive study using the Rutter A2 (parent) Scale to determine psychological morbidity. This questionnaire comprises 31 questions that were answered by a parent of the 206 subject children (ages 6 to 13 years). We selected subjects based on a multistage, randomized selection of 8 Palestinian villages and their households in the southern region of Bethlehem, West Bank. We used the Gaza Socioeconomic Adversities Questionnaire to determine differences in economic status among families. RESULTS: For all families interviewed, the father was employed, none were receiving financial assistance, and all but 1 owned their own house. The results of the Rutter A2 Scale revealed a rate of psychological morbidity ("caseness") of 42.3% among Palestinian children. The rate for boys was 46.3% and for girls, 37.8%. CONCLUSIONS: The prevalence of psychological morbidity among Palestinian children in the West Bank was significantly higher (factor of 2; chi2 = 23.26, df 1, P < 0.001), relative to the level of psychological morbidity determined independently for children in Gaza during 2000. We predict that these rates will have increased substantially owing to the escalated violence that began in this region 2 months after we conducted our study. We further predict that children in Israeli settlements in the West Bank will also exhibit elevated levels of psychological morbidity, relative to their counterparts in Israel. PMID- 14763681 TI - Modafinil treatment of excessive sedation associated with divalproex sodium. PMID- 14763682 TI - Ziprasidone in Parkinson's disease psychosis. PMID- 14763683 TI - Combined oral venlafaxine and intravenous clomipramine-A: successful temporary response in a patient with extremely refractory depression. PMID- 14763684 TI - Doxepin increases serum cholesterol levels. PMID- 14763685 TI - Re: Suicide: the persisting challenge. PMID- 14763686 TI - Depression and a history of alcoholism. PMID- 14763687 TI - Gains in speeded information processing following clozapine treatment of schizophrenia. PMID- 14763688 TI - Problems with crystallizing phenomenology and nosology in adolescent psychiatry. PMID- 14763689 TI - Serotonin syndrome with prolonged dysphagia. PMID- 14763690 TI - Imaging bones of contention. PMID- 14763691 TI - Rapid bone and blood flow formation in impacted morselized allografts: positron emission tomography (PET) studies on allografts in 5 femoral component revisions of total hip arthroplasty. AB - 5 patients were revised with impaction of morselized frozen allograft and a cemented total hip arthroplasty (THA) because of loosening and osteolysis of a primary hip arthroplasty. Plain film radiographs of the stems showed stable implants in all patients 15-24 months after surgery. The clinical results were good. We used: 1) Positron Emission Tomography (PET) to evaluate vascularization and new bone formation in the allograft, 2) kinetic [18F]-fluoride PET to produce quantitative images, interpreted as new bone formation in the allograft surrounding the femur stem, 3) [15O]-water PET to quantify bone blood flow, and 4) [15O]-carbon monoxide to determine blood volume. After surgery, all patients were evaluated twice: at 1-8 days and 12 months and 3 patients were also studied at 4 months. As early as at 8 days after surgery, blood flow and bone formation had increased greatly adjacent to the allograft. At 4 months blood flow and bone formation were about the same, but activity was highest in the graft material. At 1 year after surgery, blood flow had declined to the levels of the contralateral femur diaphysis in most of the graft bed. These findings using the PET technique showed that angiogenesis and new bone formation occurred early after impaction of morselized bone allografts around the femoral component in revision THA. We found that PET is a sensitive method for evaluating neovascularization and bone formation in the graft beds. PMID- 14763692 TI - Antibiotic prophylaxis in total hip arthroplasty: effects of antibiotic prophylaxis systemically and in bone cement on the revision rate of 22,170 primary hip replacements followed 0-14 years in the Norwegian Arthroplasty Register. AB - We studied the effects of antibiotic prophylaxis, systemically and in bone cement, on the revision rate of cemented total hip arthroplasties (THAs) in data from the Norwegian Arthroplasty Register during the period 1987-2001. To have comparable groups, only THAs performed because of primary osteoarthritis, using cemented implants with documented good results, and high-viscosity cement were included. If systemic antibiotic prophylaxis had been given, only operations with cephalosporin or penicillin were selected. Cox-estimated survival relative revision risks (RR) are presented with adjustment for differences among groups in gender, age, cement brand, type of systemic antibiotic prophylaxis, type of prosthesis, type of operating room, and duration of the operation. Of 22,170 THAs studied, 696 THAs (3.1%) were revised, 440 (2.0%) for aseptic loosening and 102 (0.5%) for deep infection. We found the lowest risk of revision when the antibiotic prophylaxis was given both systemically and in the cement (15,676 THAs). Compared to this combined regime, patients who received antibiotic prophylaxis only systemically (5,960 THAs) had a 1.4 times higher revision rate with all reasons for revision as endpoint (p = 0.001), 1.3 times higher with aseptic loosening (p = 0.02) and 1.8 times higher with infection as the endpoint (p = 0.01). With the combined antibiotic regime, the results were better if antibiotics were given 4 times on the day of surgery (2,194 THAs), as compared to once (1,424 THAs) (p < 0.001), twice (2,680 THAs) (p < 0.001), or 3 times (5,522 THAs) (p = 0.02). Those who received systemic prophylaxis a single day 1, 2 or 3 times, as compared to 4 times, had a revision rate 1.8-3.5 times higher with all reasons for revision as endpoint, 1.5-3.1 times higher with aseptic loosening, and 2.7-6.8 times higher with infection. When we compared systemic prophylaxis 4 times in 1 day, no further improvement resulted in those given systemic prophylaxis for 2 days (1,928 THAs) or 3 days (717 THAs). In a subset of data including only the Charnley prosthesis, we obtained similar results. This observational study shows that the best results were recorded when antibiotic prophylaxis was given both systemically and in the bone cement, and if the systemic antibiotic was given 4 times on the day of surgery. PMID- 14763693 TI - Particle size of bone graft and method of impaction affect initial stability of cemented cups: human cadaveric and synthetic pelvic specimen studies. AB - We determined the effect of bone graft particle size and impaction technique on the initial stability of cemented acetabular cups. First, acetabular reconstructions were performed in human cadaveric pelvic bones in which type 2 AAOS cavitary defects were created. Reconstructions were made with small bone grafts (average 2 mm) produced by a bone mill or large bone grafts (average 9 mm) produced by hand with a rongeur. All chips were made from freshly-frozen femoral heads. Impaction was done using acetabular impactors and a hammer. We did a loading experiment with a gradually increasing dynamic load up to 3000 N. We used radiostereometric analysis (RSA) to determine cup stability. The cups were more stable when large bone grafts were used. Because of limitations of the cadaver model, we developed a synthetic acetabular model. For validation of this model, we repeated the experiments using small and large bone grafts. The results with both models were similar. In the synthetic model, we compared impaction with hammer and impactors with the reversed reaming technique using manual compression on the reamer. The latter method resulted in more migration. We recommend firm impaction with a hammer of large bone grafts for optimal stability of the cup. PMID- 14763694 TI - Early bead shedding of the Vitalock acetabular cup--a report on 7 cases. PMID- 14763695 TI - Living cells in 1 of 2 frozen femoral heads. AB - Allogeneic, frozen bone is now the most commonly grafted tissue (Norman-Taylor and Villar 1997). Tissue banks collect bone material according to protocols developed with the aim of maintaining osseoinductive properties of grafts as well as preventing transmission of viral or bacterial diseases (Standards from the American Association of Tissue Banks (AATB) or from the European Association for Musculo-skeletal Transplanting (EAMST)). Standard procedures include cryopreservation of tissue at -80 degrees C, which is generally considered to devitalize the bone by killing all cells present, resulting in reduced immunogenicity of the graft. The osseoinductive properties of frozen, allogeneic bone grafts have therefore mainly been attributed to the dead bone matrix, that may provide osteoblast-stimulating growth factors and other essential proteins, and/or an osteoclast substrate to direct bone remodeling (Aspenberg et al. 1996, Kingsmill et al. 1999). Recently however, it was suggested that some cells in bone biopsies may survive standard bone bank freezing procotols. It is unclear whether vital cells are present in other bone banks and whether these cells can contribute to the clinical outcome of frozen allogeneic bone grafting. In this report, we show that frozen bone biopsies, obtained from the Erasmus Medical Center bone bank may contain living cells that can be cultured in vitro. These cultured cells were found to originate from the donor by genotyping. PMID- 14763696 TI - Tranexamic acid reduces blood loss and blood transfusions in primary total hip arthroplasty: a prospective randomized double-blind study in 40 patients. AB - INTRODUCTION: We performed a prospective, randomized, double-blind study on 40 patients scheduled for primary total hip arthroplasty due to arthrosis or osteonecrosis to determine the effect of tranexamic acid on per- and postoperative blood losses and on the number of blood transfusions needed. PATIENTS AND METHODS: 40 patients were randomized to tranexamic acid (10 mg/kg given as a bolus intravenous injection, followed by a continuous infusion of 1 mg/kg/hour for 10 hours) or placebo (20 mL saline given intravenously) 15 minutes before the incision. We recorded the peroperative and postoperative blood losses at removal of the drain 24 hours after the operation and the number of blood transfusions. RESULTS: Patients receiving tranexamic acid had a mean peroperative blood loss of 480 mL versus 622 mL in patients receiving placebo (p = 0.3), a postoperative blood loss of 334 mL versus 609 mL (p = 0.001), a total blood loss of 814 mL versus 1231 mL (p = 0.001) and a total need for 4 blood transfusions versus 25 (p = 0.04). No patient in either group had symptoms of deep venous thrombosis, pulmonary embolism or prolonged wound drainage. INTERPRETATION: Transemic acid is effective in reducing the postoperative blood loss, the total blood loss and the need for blood transfusion in primary total hip arthroplasty. PMID- 14763697 TI - The effect of mixing on gentamicin release from polymethylmethacrylate bone cements. AB - We compared the release of gentamicin from 6 different commercially available, antibiotic-loaded PMMA bone cements used for vacuum- and hand-mixed cement using a Cemvac vacuum mixing system. We also measured the release of gentamicin after manual addition of the antibiotic to different commercial, unloaded bone cements after hand-mixing. The porosity of cements was reduced in all vacuum-mixed cements, as compared with hand-mixed cements, concurrent with a statistically significant reduction (3 of 6) or increase (1 of 6) in the total amounts of gentamicin released. The total gentamicin release was studied in 3 of the brands after manual addition and mixing of the antibiotics. We found that the release of antibiotics was lower than in samples made from industrial mixing. In conclusion, the manual addition and mixing of gentamicin in PMMA bone cements leads to a lower release of antibiotics than that in corresponding commercially available antibiotic-loaded cements, while vacuum-mixing only leads to a minor reduction in antibiotic release, as compared to hand-mixing. PMID- 14763698 TI - The precision and influence of rotation for measurements of bone mineral density of the distal femur following total knee arthroplasty: a methodological study using DEXA. AB - We evaluated the feasibility of DEXA (Norland XR-26 mark II) for quantitative measurements of bone mineral density (BMD) in the lateral plane of the distal femur after total knee arthroplasty (TKA). BMD was measured in 5-6 regions of interest (ROI) in close relation to the femoral component. In an in vitro study using 3 different distal femur phantoms, we found that the precision was affected by rotation of the distal femur. When BMD measurements were repeated within a range of motion of 40 degrees, 20 degrees, and 0 degrees, the coefficient of variation (CV) was approximately 15%, 10%, and 0.6%, respectively. We found that the use of bone cement for implant fixation had no effect on the level of BMD. Double measurements performed in 28 patients gave average CV values of 3.3%, 3.0%, and 2.6% for the uncemented Duracon, and Interax femoral components and the cemented AGC components, respectively. Our in vivo average CV measurements of BMD of the distal femur after TKA were on a level, suitable for repeated BMD measurements in prospective studies, which evaluate adaptive bone remodeling of the distal femur after cemented and uncemented TKA. PMID- 14763699 TI - Temporal changes in posterior laxity after isolated posterior cruciate ligament injury: 35 patients examined by stress radiography and MRI. AB - We examined the temporal changes in posterior laxity and MRI findings in acute isolated posterior cruciate ligament (PCL)-injured knees. 35 patients with acute isolated PCL injuries who attended our clinic within 10 weeks of an injury underwent nonoperative treatment, and were chronologically evaluated for posterior laxity and MRI. We divided them in 3 groups, as regards the evolution of laxity and MRI findings. In 20 patients, the degree of posterior laxity was slight throughout the observation periods. In 9 patients, who had severe posterior laxity on their initial visit, the laxity had improved markedly 6 months after injury and did not deteriorate. In 6 patients the severe posterior laxity did not improve during the study. 4 patients whose PCL on MRI was seen as a continuous swollen band having high signal intensity and showed consistently good stability or even an improvement. We conclude that acute isolated PCL injuries with a continuous swollen band on MRI should be treated conservatively for at least 6 months after injury, even if the posterior laxity is severe initially. PMID- 14763700 TI - Fluorescence immunohistochemistry and confocal scanning laser microscopy: a protocol for studies of joint innervation. AB - We have developed a reliable technique for labeling and examining neural structures in soft tissues associated with articular joints and have tested it in human wrist joints under various specimen-related conditions. The labeling protocol employs an immunohistochemical process with a panneuronal marker (PGP 9.5) as the primary antibody and Alexa Fluor 488 as the fluorescing secondary antibody. Imaging was done using a confocal laser scanning microscope, which produced exceptionally detailed three-dimensional images of nerve endings and transiting nerve fibers from thick sections of wrist joint ligaments obtained from human cadavers. The protocol provided a practical postmortem window for specimen acquisition and processing without significant apparent worsening of image quality. The images produced are resistant to fading with repeated exposure to a fluorescent light source, which gives many opportunities for observation. Background staining is minimal, producing high contrast labeling of target tissues, which, in turn, enhances image analysis. PMID- 14763701 TI - Anterior knee pain in the young patient--what causes the pain? "Neural model". AB - Anterior knee pain in young patients is the commonest type of knee disorder in clinical practice. However, the pathogenesis of this condition is unknown. On the basis of our recent research, we suggest a "neural model". In our view, hyperinnervation in the lateral retinaculum, mainly nociceptive substance P positive nerves induced by the release of neural growth factor, is involved in the pathogenesis of anterior knee pain. We hypothesize that periodic short episodes of ischemia may trigger neural proliferation. PMID- 14763702 TI - No difference between daily and weekly pin site care: a randomized study of 50 patients with external fixation. AB - INTRODUCTION: We investigated whether there were any differences in the frequency and severity of pin site infections by performing pin site care daily or once a week. We studied patients operated on for gonarthrosis by the hemicallotasis technique, using hydroxyapatite-coated pins in the metaphyseal bone and standard pins in the diaphyseal bone. PATIENTS AND METHODS: 50 patients were prospectively randomized to daily (n = 27) or weekly (n = 23) pin site care. We evaluated pin sites, the occurrence of pain (VAS), the use of antibiotics and analgesics and complications every week. Bacterial cultures were taken from each pin site at 1, 6 and 10 weeks and from the pins on removal. RESULTS: We found no differences between daily or weekly pin site care as regards the frequency and severity of pin site infections, pain, or the use of antibiotics and analgesics. Grade I infections (Checketts-Otterburns classification) occurred around 11% of the pins and grade II infections around 4%. 70% of the bacterial cultures were negative. The most frequent bacteria were coagulase negative staphylococcus and corynebacterium. Antibiotics were given an average of 47 days. More problems occurred around the proximal pins. 5/200 (all proximal) pins were clinically loose on removal. INTERPRETATION: Pin site care once a week seems appropriate. PMID- 14763703 TI - Effect of distraction frequency on bone formation during bone lengthening: a study in chickens. AB - We compared the effects of two distraction frequencies on bone formation during tibial lengthening by evaluating radiographs, bone mineral density, and histological findings. In 15 mature White Leghorn chickens, both tibiae were distracted at a rate of 0.75 mm/day for 10 days. The distraction frequency was 2 steps (0.375 mm/12 hour) by hand on the right side and 120 steps (0.00625 mm/12 min) by autodistractor on the left. Serial radiographs showed faster bone formation on the 120-step side than on the 2-step side. Bone mineral density on the 120-step side was also higher than that on the 2-step side at all times. On the 2-step side, endochondral ossification was marked in the early stage of distraction; then intramembranous ossification became the main mechanism of bone formation. On the 120-step side, however, intramembranous bone formation predominated throughout the study. Our findings support the contention that, at least in skeletally mature chickens, an increase in the distraction frequency improves osteogenesis during bone lengthening. PMID- 14763704 TI - Increased mortality after fracture of the surgical neck of the humerus: a case control study of 253 patients with a 12-year follow-up. AB - BACKGROUND: Several studies have shown a higher mortality rate in patients with osteoporosis-related fractures of the hip and vertebrae. METHOD: In 1999, we did a long-term follow-up case-control mortality study of 253 patients, mean age 72 years, who had sustained a fracture of the surgical neck of the humerus in 1987. RESULTS: We found a higher mortality in fracture patients giving at end point a cumulative survival difference of 16%. The median survival time was 8.9 years in patients and 12 years in controls (p = 0.005). The mortality rate was higher in men during the first 3 years after fracture and fewer than half of the male patients survived this period. The median survival time was 6.5 years in male patients and 12 years in their male controls (p = 0.02). The mortality was only slightly higher in women (p = 0.06). INTERPRETATION: Cardiovascular disease and malignancy were the commonest causes of death in both groups. We could not explain the higher mortality rate in patients with a fracture of the surgical neck of the humerus. PMID- 14763705 TI - The relation of the coracoclavicular ligament insertion to the acromioclavicular joint: a cadaver study of relevance to lateral clavicle resection. AB - Resection of the lateral end of the clavicle is a common procedure for arthrosis of the acromioclavicular joint (AC-joint). However, no anatomical data on the distance between the insertions of the coracoclavicular ligaments and the AC joint have been reported. In 36 cadaver shoulders (18 male), we studied the relation between the AC-joint and the insertions of the joint capsule, trapezoid and conoid ligaments. The distance from the AC-joint to the medial end of its capsule was, on average, 0.7 cm (0.4-0.9) cm in women and 0.8 (0.4-1.2) cm in men. In women, the trapezoid ligament began, on average, at 0.9 (0.4-1.6) cm and ended at 2.4 (2.0-2.8) cm and in men, it began at 1.1 (0.8-1.6) cm and ended at 2.9 (2.1-3.8) cm medial to the AC joint. The corresponding figures for the conoid ligament were 2.6 (2.0-3.7) cm and 4.7 (3.9-6.2) cm. A resection of 1 cm of the lateral clavicle detaches 8%, a resection of 2 cm 60% and a resection of 2.5 cm 90% of the trapezoid ligament. We recommend a maximum resection of 1 cm of the lateral clavicle because a resection of 2 cm or more may cause postoperative AC joint instability and related pain. PMID- 14763706 TI - Practical assessment of rotator cuff muscle volumes using shoulder MRI. AB - Reliable quantitative assessment of rotator cuff (RC) muscle volumes can be done by reconstructing multiple MRI images of the entire shoulder. However, an equally reliable, but less time-consuming, method is needed for clinical practice. We compared the only method reported for estimation of volume with a new simple MRI technique. Both methods were validated by multiple MRI image reconstruction. We performed MRI scans of 10 cadaver shoulders and determined the cross-sectional areas of RC muscles with two methods, using image analyzing software. In Method 1, the cross-sections were determined on a single image, previously described as a Y-shaped image. In Method 2, the cross-sections were calculated from two images: the Y-shaped and an image located medially, twice the distance from the glenoid articular surface compared to the Y-shaped image. We compared the results of these two methods with the volume of multiple MRI image reconstruction, which took into account all images consisting of RC cross-sections. Pearson correlations for Method 1 were 0.96, 0.94 and 0.75, and for Method 2, 0.96, 0.97 and 0.93 for the supraspinatus, infraspinatus/teres minor and subscapularis muscle volumes when compared with the volumes determined by the multiple image reconstruction method. The Bland-Altman method showed better agreement with multiple MRI image reconstruction, using Method 2, to determine supraspinatus, infraspinatus, and subscapularis muscle volumes (p < 0.001 for each). The mean intra- and inter-observer variabilities of Method 1 was 3.9% and 2.9% and that of Method 2, 3.0% and 1.7%, respectively. Both methods can be used for quantitative assessment of RC muscle volumes. However, Method 2, using two easily reproducible MRI images is more accurate for the evaluation of the supraspinatus and infraspinatus/teres minor muscles and particularly for the subscapularis muscle. PMID- 14763707 TI - In vitro osteoblast-like cell metabolism in spondylodesis--a tool that may predict fusion capacity: a prospective study in 50 patients with a 1-year follow up. AB - In vitro cultures of human primary osteoblast-like cells provide a model for studying cellular mechanisms associated with human bone biology. We investigated in vitro osteoblast-like cell metabolism as a method for predicting the occurrence of spinal fusion in the individual patient. A bone biopsy was taken from the iliac crest of 50 patients, median age 49 (23-77) years, who were undergoing lumbar spine fusion. First-passage osteoblast-like cells were established by the bone-tissue-explant method. We then estimated 3H-thymidine incorporation, alkaline phosphatase activity and procollagen I production. Fusion rates were evaluated at the 1-year follow-up. Primary human osteoblast-like cell cultures showed an age-dependent decline in their capacity for cellular outgrowth and expression of alkaline phosphatase, which suggested a useful biological response pattern of the osteoblast culture. However, such cultures were unsatisfactory as an in vitro tool for predicting fusion capacity. PMID- 14763708 TI - X-linked spondyloepiphyseal dysplasia tarda: molecular cause of a heritable disorder associated with early degenerative joint disease. AB - Spondyloepiphyseal dysplasia tarda (SEDT) is a genetically heterogeneous disorder often associated with the early onset of osteoarthrosis. The X-linked recessive form (SEDL) affects men and is characterized by reduced height, arm span exceeding total height, and barrel chest deformity. The radiographic phenotype comprises a hump-shaped deformity of vertebral bodies and mild epiphyseal dysplasia of the femoral head associated with early signs of hip arthrosis. The disorder is caused by mutations in the SEDL (or sedlin) gene on Xp22.12-p22.31. In 4 male patients from a German family, we identified a new nonsense mutation in SEDL exon 4 (C74A). The carrier status for the mutation could be confirmed in 2 women of the family, both of whom show no obvious signs of bone and joint diseases. SEDT should be kept in mind as a differential diagnosis in men with early "primary" bilateral osteoarthrosis. PMID- 14763709 TI - No augmentation of morselized and impacted bone graft by OP-1 in a weight-bearing model. AB - INTRODUCTION: Impacted morselized bone allograft is thought to be remodeled to a great extent. We have previously shown that load-bearing increases the remodeling of impacted morselized bone allografts in a rabbit knee prosthesis model. Bone Morphogenic Proteins (BMPs) also stimulate bone formation and bone allograft remodeling. In this study, our aim was to determine whether it was possible to increase further the remodeling of impacted morselized bone allografts by combining load-bearing with a BMP. EXPERIMENT: A solution of Osteogenic Protein-1 (OP-1, also called BMP-7) was added to freeze-dried bone allograft chips before impaction grafting in our rabbit knee prosthesis model. 23 skeletally mature rabbits received an uncemented tibial knee prosthetic component inserted into a bed of impacted morselized bone allograft. 12 rabbits were given OP-1-treated allograft (50 microg OP-1 per gram allograft), and 11 rabbits vehicle-treated allograft. Each rabbit received mean 0.53 g graft. The rabbits were killed after 3 or 6 weeks and the grafted region was examined by histomorphometric assessment of the volume fraction of newly formed bone and remaining graft. RESULTS AND INTERPRETATION: We found that OP-1 did not increase the bone density (graft plus new bone) to any substantial extent. However, we can not exclude that this might be due to a carrier problem, since the OP-1 was added as a solution directly to the dry graft. PMID- 14763710 TI - Botulinum A toxin for treatment of lower limb spasticity in cerebral palsy: gait analysis in 49 patients. AB - BACKGROUND: Injection of botulinum type A toxin is a new treatment for spasticity. PATIENTS AND METHODS: We evaluated the effect of botulinum A toxin (BTX-A) in the lower limb muscles of patients having cerebral palsy. We tested 49 patients before and, on average, 4 (2-9) months after giving the toxin. The evaluation included 3-dimensional computerized gait analysis, changes in mobility level, using the Gillette Functional Assessment Questionnaire, and gastrocnemius muscle bulk, using ultrasonographic measurements. RESULTS: The patients were divided into 3 groups, according to the site of BTX-A administration (hamstrings, gastrocnemius and multilevel). Those who were injected in the hamstrings showed a significant improvement in only the maximum knee extension angle during the gait cycle. Those with spastic equinus who were injected in the gastrocnemius muscle responded better than the other groups. The ankle angle on the initial contact, terminal stance and pre-swing, maximum dorsiflexion, ankle range of motion, per cent of single support and gait velocity improved significantly. Overall, the patients showed significant improvements in motor skill performance and functional health. INTERPRETATION: Our findings indicate that botulinum type A toxin can be given as an adjuvant to conservative treatment of patients with cerebral palsy. PMID- 14763711 TI - The Short Musculoskeletal Function Assessment Questionnaire (SMFA): cross cultural adaptation, validity, reliability and responsiveness of the Swedish SMFA (SMFA-Swe). AB - The Short Musculoskeletal Function Assessment Questionnaire (SMFA) is designed to measure the functional status of patients with a broad range of musculoskeletal injuries and disorders. It has previously been validated for an American population. We have translated the SMFA into Swedish and tested the translated version (SMFA-Swe) as regards validity, reliability and responsiveness. Acute and elective cases (n = 298) were included in the study. The Swedish version of the SF-36 was used in the validation. We found that the SMFA-Swe was easy to use, that its reliability (internal consistency and stability) was good, that it correlated well with the SF-36 physical scores and that it was also sensitive to changes in musculoskeletal function over time. Orthopedic studies frequently use disease-specific outcome measures because they are sensitive to the disorder studied. However, there may also be a need to evaluate function in groups of patients with different or multiple musculoskeletal disorders and, in such situations, the SMFA can be useful. We conclude that the SMFA-Swe is a valid instrument and can be used in clinical research as well as clinical practice when focusing on patients with various musculoskeletal disorders. PMID- 14763712 TI - Upper tibial physeal fracture--a case report. Proposed mechanism of injury and classification. PMID- 14763713 TI - Remodeling of the tibia after grafting of a large cavity with particulate bioactive glass-hydroxylapatite--case report on treatment of fibrous dysplasia with 13 years' follow-up. PMID- 14763714 TI - Cox inhibitors and bone healing. PMID- 14763715 TI - Occupational therapy for children with cerebral palsy: a systematic review. AB - OBJECTIVE: Occupational therapy (OT) for cerebral palsy focuses on the development of skills necessary for the performance of activities of daily living. The aim of this systematic review was to determine whether OT interventions improve outcome for children with cerebral palsy (CP). METHODS: An extensive search in MEDLINE, CINAHL, EMBASE, AMED and SCISEARCH was performed. Studies with controlled and uncontrolled designs were included. Six intervention categories were distinguished and individually analysed using a best-evidence synthesis. This synthesis is based on the type of design, the methodological quality, the type of outcome measures and the statistical significance of the findings. RESULTS: Seventeen studies were included in this review, seven of which were randomized controlled trials (RCTs). One RCT had a high methodological quality. The analyses resulted in insufficient evidence of the efficacy of occupational therapy in all intervention categories, due to the low methodological quality of studies presenting statistically nonsignificant results. CONCLUSION: Despite the reasonable number of studies identified, the inconclusive findings regarding the efficacy of occupational therapy for children with cerebral palsy may be a reflection of the difficulties in efficacy research in OT for children with CP. Future research should critically reflect on methodological issues. PMID- 14763716 TI - A systematic review of the methodological quality and extent to which evaluation studies measure the usability of orthopaedic shoes. AB - OBJECTIVE: To determine the methodological quality of studies evaluating orthopaedic shoes and orthopaedic shoe provisions. To what extent do studies evaluating orthopaedic shoes prescribed for patients with degenerative disorders of the foot, rheumatoid arthritis, diabetes mellitus and neurological foot disorders, focus on the aspects of the International Organization for Standardization (ISO) definition of usability, i.e., effectiveness, efficiency, satisfaction, and context of use? DESIGN: Systematic review. METHODS: A systematic literature search was performed to identify randomized controlled trials (RCTs) concerning orthopaedic shoes and orthopaedic shoe inserts. The methodological quality of the studies was assessed independently by two raters, based on the 19 items of the 'Maastricht-Amsterdam criteria list'. The studies were assessed against the parameters of the ISO definition of usability. RESULTS: Eleven RCTs were included. The methodological scores ranged from 8 to 14 out of 19 possible points. Eleven studies focused on the effectiveness of the orthopaedic shoes and orthopaedic shoe inserts, two of which reported outcome measures and conclusions related to the efficiency of the studied orthopaedic shoes and orthopaedic shoe inserts. Four studies reported some form of patient satisfaction and only three studies paid attention to the context of use. CONCLUSIONS: The methodological quality of the RCTs as assessed according to the 19 different criteria varied considerably. The present review shows that current scientific literature concerning the usability of orthopaedic shoes focuses mainly on effectiveness at the expense of the other domains of usability, i.e., efficiency, satisfaction and context of use. PMID- 14763717 TI - Outcomes of progressive resistance strength training following stroke: a systematic review. AB - OBJECTIVE: To determine whether progressive resistance strength training reduces impairments, activity limitations and participation restrictions after stroke. METHODS: Electronic databases were searched to find trials conducted from 1966 to 2002. Articles were assessed independently by two reviewers according to the following inclusion criteria: (1) population: adults with stroke, (2) intervention: progressive resistive strength training in isolation, and (3) outcomes: changes in body function, physical activity or societal participation. RESULTS: From the 350 articles initially identified, eight met the criteria for detailed review. Three were randomized controlled trials, with the remainder being single-case time-series analyses or pre-post trials. The five trials that measured impairments of muscle strength showed positive outcomes for progressive resistance strength training, with large effect sizes (d = 1.2-4.5). Few negative effects of strength training were reported, and these were minor. Only three of the eight trials that measured activity limitations reported improvements in activities such as walking and stair climbing. The effects of strength training on societal participation could not be determined due to insufficient data. CONCLUSIONS: There is preliminary evidence that progressive resistance strength training programmes reduce musculoskeletal impairment after stroke. Whether strengthening enhances the performance of functional activities or participation in societal roles remains unknown. PMID- 14763718 TI - Evaluation of a multiprofessional community stroke team: a randomized controlled trial. AB - OBJECTIVES: There is inconclusive evidence of the effectiveness of stroke rehabilitation by a community stroke team. The aim was to evaluate a specialist multiprofessional team in a community setting. DESIGN: Randomized controlled trial. SETTING: Community. PARTICIPANTS: Stroke patients and their informal carers who were referred to receive rehabilitation from a community stroke team. OUTCOME MEASURES: Barthel Index, Extended Activities of Daily Living Scale (EADL), General Health Questionnaire (GHQ-12) by patient and carer, Carer Strain Index (CSI), Euroquol, knowledge of stroke and satisfaction with services six months after recruitment. RESULTS: There were no significant differences between patients who received rehabilitation from community stroke team (n = 189) and those who received routine care (n = 232) in their independence in activities of daily living, mood, quality of life or knowledge of stroke. The patients in the community stroke team group were significantly more satisfied with the emotional support they had received (p < 0.01). There were no significant differences between the groups in satisfaction with practical help or overall satisfaction. Carers of patients in the community stroke team were under significantly less strain than carers in the routine care group (p < 0.04). Carers of patients in the community stroke team group were significantly more satisfied with their knowledge of stroke (p < 0.01) and were more satisfied overall (p < 0.01). CONCLUSIONS: The patients treated by the community stroke team were more satisfied with the emotional support they received and had equivalent outcomes in terms of independence in activities of daily living and mood. Their carers were under less strain and were more satisfied with their knowledge of stroke recovery, the emotional support they received and overall satisfaction with services. The results support the provision of rehabilitation by a community based specialist multiprofessional team. PMID- 14763719 TI - Neurophysiological and behavioural adaptations to a bilateral training intervention in individuals following stroke. AB - OBJECTIVE: To examine the neurophysiological and behavioural adaptations to a bilateral training protocol in individuals with hemiparesis due to stroke. DESIGN: Subjects performed daily practice sessions of three specific upper limb tasks over a four-week period. Performance of the tasks was recorded on video for later analysis over the duration of the study. For the first week of the training period the tasks were practised with the impaired upper limb. Only then, with a random start day between 8 and 13, the tasks were practised using a bilateral training protocol. Functional assessments of the affected upper limb and neurophysiological investigations of contra- and ipsilateral pathways from the affected and nonaffected cortical hemispheres were completed prior to the start of the intervention, at the end of the unimanual practice period, and at the conclusion of the bimanual practice period. SUBJECTS: Six individuals between one month and four years post stroke. INTERVENTIONS: Unimanual and bimanual training protocols. MAIN MEASURES: Task performance with the affected limb only; motor impairment of the affected upper limb; ipsi- and contralateral corticospinal pathway excitability. RESULTS: Baseline task performance varied markedly between subjects. Most individuals demonstrated little, if any, additional beneficial effect of bimanual practice on task performance. Results of neurophysiological investigations were inconsistent between subjects. CONCLUSIONS: Short-term bilateral training following unilateral training may have limited effectiveness in enhancing upper limb motor performance in acute and chronic individuals poststroke. The neural mechanisms associated with bilateral activation therapies remain uncertain. PMID- 14763720 TI - The adjunctive role of mental practice in the rehabilitation of the upper limb after hemiplegic stroke: a pilot study. AB - OBJECTIVE: To assess the feasibility and practicalities of using the technique of mental practice as an adjunct in the rehabilitation of the upper limb following stroke. DESIGN: A series of single-case studies. SETTING: A stroke rehabilitation unit in Belfast. SUBJECTS: Fourteen patients admitted for rehabilitation of their first stroke: six men and four women, aged 45-81 between 10 and 176 days post stroke. INTERVENTION: Each patient underwent a single-case design, with two weeks baseline, two weeks intervention and one week withdrawal. The intervention consisted of structured daily mental practice sessions of a reach and grasp task, in addition to their usual therapy. MAIN OUTCOME MEASURES: The upper limb component of the Motricity Index was used to grade motor activity sequentially across the timescale of the study. RESULTS: Of the 14 patients recruited, four withdrew and 10 completed the study. Nine showed improvement in upper limb Motricity Index score with mental practice as measured by the two-band standard deviation method. One of these cases demonstrated an unstable baseline such that changes could not be attributed to intervention. CONCLUSIONS: This pilot study suggests that mental practice may be useful as an adjunct to physiotherapy after stroke. PMID- 14763721 TI - Evaluation of a pilot service designed to provide support following stroke: a randomized cross-over design study. AB - OBJECTIVE: To evaluate a day service for people aged 18-55 years who had a stroke. DESIGN: A randomized cross-over study design was used, randomly allocating individuals to attend the service for six months followed by a period of no attendance for six months. SETTING: A day service pilot project was launched in Cardiff in July 1995 for people who were aged between 18 and 55 years and had a stroke. It met one day a week. SUBJECTS: Twenty-six participants were recruited to the study between June 1998 and February 2000. Their mean age was 48 years (SD = 7). INTERVENTIONS: The service aimed to offer participants the opportunity to identify and pursue meaningful and realistic opportunities within the community. A range of activities occurred at the service including creative activities and social outings. MAIN OUTCOME MEASURES: The Barthel ADL Index, Extended ADL Scale, Nottingham Leisure Questionnaire, Short Form 36, the Hospital Anxiety and Depression Scale, the Canadian Occupational Performance Measure, the Role Checklist and the Semantic Differential Self Concept Scale were used to assess the outcomes from the service. RESULTS: Attending the service increased occupational performance and satisfaction with performance but there was no evidence that depression and anxiety were reduced or that quality of life and self-concept were improved. CONCLUSION: Although there were some gains from attending the service there were also many unmet needs. Further research is required to continue to identify how best to meet the needs of individuals post stroke under retirement age. PMID- 14763722 TI - Efficacy of vestibular rehabilitation on chronic unilateral vestibular dysfunction. AB - OBJECTIVE: To assess the efficacy of vestibular rehabilitation exercises on patients with chronic unilateral vestibular dysfunction. DESIGN: Prospective study. SETTING: Physical Medicine and Rehabilitation Clinic and Otolaryngology Clinic of a tertiary referral hospital. SUBJECTS: One-hundred and twenty-five patients with unilateral chronic vestibular dysfunction were included in the study. INTERVENTIONS: Eight-week, two-staged (clinic and home) vestibular rehabilitation programme with components of Cawthorne-Cooksey and Norre exercises was applied. MAIN OUTCOME MEASURES: Dizziness Handicap Inventory (DHI) and visual analogue scale (VAS) were completed three times (at the beginning, end of the second week and end of the treatment). RESULTS: Data for 112 patients in the first stage and 93 patients in the second stage were evaluated because of insufficient compliance of the other patients. The mean DHI score was decreased from 50.42 +/- 24.12 points to 21.21 +/- 15.97 points (p < 0.001) at the end of first two weeks, and to 19.93 +/- 19.33 points at the end of the whole treatment. The mean VAS score was decreased from 5.87 +/- 2.27 to 2.02 +/- 1.75 (p < 0.001) at the end of second week, and to 1.51 +/- 1.29 at the end of eighth week. In respect to both VAS and DHI scores, improvement was noted in 67 patients (77.4%). Age, gender and disability level had no predictive value about therapy outcome. CONCLUSIONS: There was a fast recovery in the supervised exercise session, whereas there was no significant difference in the home exercise session. These findings suggest that either supervised exercise is better than home exercise or that 10 supervised sessions are sufficient to get the end result. PMID- 14763723 TI - Comparison of two different rehabilitation programmes for thrust plate prosthesis: a randomized controlled study. AB - OBJECTIVE: Weight bearing after total hip arthroplasty is postponed in order to prevent early loosening, but this negatively affects the rehabilitation programme. For the force transfer characteristics of thrust plate prosthesis (TPP), a new type of hip prosthesis used without cement is similar to the normal hip. We evaluated the possibilities of early weight bearing after TPP by comparing early partial with early full weight bearing. DESIGN: Randomized controlled study. SETTING: Department of orthopaedics and traumatology in a university hospital. SUBJECTS: Sixty hips of 51 patients who underwent total hip arthroplasty with TPP were randomly assigned into two groups. INTERVENTIONS: Both groups received accelerated rehabilitation programmes: group 1 with early partial weight bearing and group 2 with early full weight bearing. MAIN OUTCOME MEASURES: Patients were evaluated by a blind observer preoperatively, at three months after surgery by clinical (measurement of range of hip motion (universal goniometry), muscle strength (Manual Muscle Test), functional test (6-minute walk test), hip function (Harris Hip Scoring System)) and radiographical parameters and one year after surgery by clinical (Harris Hip Scoring System) and radiographical parameters. RESULTS: Group 2 performed transfer activities earlier, had more walking distance at the time of discharge and shorter hospital stay than group 1. At three months, Harris Hip Score, muscle strength, 6-minute walk test, and duration of crutch use were significantly (p < 0.05) in favour of group 2. None of the patients in either group showed signs of loosening one year after the operation. CONCLUSIONS: These results suggest that patients with TPP can tolerate an accelerated rehabilitation programme with early weight bearing and will gain the goals of rehabilitation earlier. PMID- 14763724 TI - Community rehabilitation for older adults with osteoarthritis of the lower limb: a controlled clinical trial. AB - OBJECTIVE: To examine the effectiveness of a 12-month community-based water exercise programme on measures of self-reported health and physical function in people aged over 60 years old with knee-hip osteoarthritis (OA). DESIGN: A quasi experimental design consisting of an exercise group and an age-matched control group. SETTING: Public community swimming pool in Sheffield, UK. SUBJECTS: One hundred and six community-dwelling sedentary older people, with confirmed knee hip osteoarthritis, enrolled in an experimental controlled trial for 12 months. Sixty-six subjects in the exercise group were offered a water-exercise programme. Forty age-matched, nonexercising, 'control' subjects received monthly education material and quarterly telephone calls. INTERVENTIONS: Participants in the exercise group were asked to attend two exercise sessions a week of 1 hour duration led by specially trained swimming instructors. MAIN MEASURES: Primary outcome measure was the disease-specific Western Ontario and McMaster University Osteoarthritis Index (WOMAC). Secondary outcomes included a battery of performance-based physical function tests. RESULTS: Adherence to exercise averaged 70% (+/- 14%) over the year: 77% of the exercising subjects and 89% control subjects completed both pre- and post-outcome measures. After one year, participants in the exercise group experienced a significant improvement in physical function (4.0 +/- 9.1 versus -0.4 +/- 7.3 units; 95% confidence interval (CI) 0.96-7.96, p < 0.05) and reduction in the perception of pain (1.3 +/- 3.7 versus 0.2 +/- 2.5 units; 95% CI -0.19-2.52, p < 0.05) compared with the control group, as measured by the WOMAC Osteoarthritis Index. In addition, the exercise group performed significantly better in the ascending and descending stairs tests (p < 0.05), had significantly greater improvements in knee range of movement (p < 0.01) and hip range of movements (p < 0.005). There were no significant differences in the two groups for quadriceps muscle strength and psychosocial well-being (Arthritis Impact Measurement Scales 2 questionnaire). CONCLUSIONS: Older people with knee/hip osteoarthritis gained modest improvements in measures of physical function, pain, general mobility and flexibility after participating in 12 months of community-based water exercise. PMID- 14763725 TI - Validation of a lateropulsion scale for patients recovering from stroke. AB - OBJECTIVE: To determine the validity and reliability of a clinical scale for assessing lateropulsion following stroke. DESIGN: Serial observational study of Lateropulsion Scale scores. SETTING: Inpatient stroke rehabilitation unit. SUBJECTS: A convenience sample of 85 patients examined 19 +/- 2 SEM days post stroke. MAIN OUTCOME MEASURES: An empirically derived 17-point Lateropulsion Scale was used to assess and follow postural responses to rolling, sitting, standing, transferring and walking. Intraclass correlation coefficients were calculated by having patients evaluated twice by their primary physical therapist (days 1 and 3), and once by an alternate physical therapist (day 2). Concurrent validity was estimated by computing Spearman's rank order correlations between the lateropulsion score and other markers for motor control dysfunction: Fugl Meyer balance subscore, the Functional Independence Measure (FIM) mobility subscore, and length of rehabilitation hospital stay. RESULTS: Inter-rater and intra-rater reliability were r = 0.93 (p < 0.001) and r = 0.94 (p < 0.05), respectively. Concurrent validity estimates showed the initial lateropulsion score to be correlated with the Fugl-Meyer balance subscore (r = -0.57 p < 0.001), with the admission and discharge FIM mobility subscores (r = -0.56, p < 0.0001 and r = -0.58, p < 0.0001), respectively, and with length of rehabilitation hospital stay (r = 0.6, p < 0.0001). CONCLUSIONS: The Lateropulsion Scale is both a reliable and a valid assessment of lateropulsion following stroke. PMID- 14763726 TI - Constraint-induced movement therapy: time for a little restraint? AB - Our aim was to consider some issues surrounding constraint-induced movement therapy (CIMT), and in particular, its theoretical basis, effectiveness, utility and composition. We examined selected articles and related publications concerning CIMT. Considerable evidence from case studies and case series has accumulated but only a limited number of randomized controlled trials (RCTs) exist. The two most positive RCTs represent a combined total of 15 people undergoing CIMT. Other issues include: how analogous deafferentation of an upper limb in monkeys is to cerebral infarcts in humans; teasing out the active components of CIMT; a need for replication by groups not already strongly associated with CIMT; and patient/therapist acceptability. CIMT may hold considerable promise, but independent, large-scale, multicentre RCTs comparing its effectiveness with conventional therapy of equal intensity are required, as is the consideration of some associated issues. PMID- 14763727 TI - Rapid determination of benzo(a)pyrene in olive oil samples with solid-phase extraction and low-pressure, wide-bore gas chromatography-mass spectrometry and fast liquid chromatography with fluorescence detection. AB - Benzo(a)pyrene (B(a)P) was extracted from olive oil using solid-phase extraction on columns filled with Florisil and Nucleoprep C18. The extracts were analyzed with GC-MS, using standard capillary column and low-pressure wide-bore column (LP GC-MS), as well as with HPLC on standard column and short donor-acceptor complex chromatography (DACC) column. Quantitation was done with isotope dilution method (GC-MS and LP-GC-MS) or with internal standard benzo(k)fluoranthene (HPLC). Limits of detection were 1 ng/g for GC-MS on standard column, 1.6 ng/g on LP column, 0.5 ng/g for HPLC on standard column, and 0.3 ng/g on DACC column, respectively. The applied extraction method allowed handling over 50 samples per day and assured recovery over 80%. Matrix solid-phase dispersion, tried as an alternative isolation method, appeared less advantageous. Fast chromatographic methods (LP-GC-MS and HPLC on DACC) made it possible to reduce analysis time to 8 and 5 min, respectively. The method was applied to routine analysis of B(a)P in olive oil samples. PMID- 14763728 TI - Determination of catechins by means of extraction with pressurized liquids. AB - The technique of extraction with pressurized liquids is applied for extraction of catechin and epicatechin in tea leaves and in grape seeds. The extracts obtained are then analyzed by HPLC. A comparison has been made of the recoveries obtained employing extraction by magnetic stirring, ultrasound-assisted extraction, and extraction with pressurized liquids. In the three extraction systems, four different pure solvents were utilized, namely water, methanol, ethanol, and ethyl acetate. Methanol produced the best results. For this comparison, an initial step was to check the stability of catechins during the process of extraction using pressurized liquids at high temperature (100-200 degrees C). It has been confirmed that recoveries of these two compounds begin to fall, to below 95%, at 130 degrees C and above. Pressurized liquid extraction using methanol as solvent, produces results, in terms of recovery of catechin and of epicatechin, notably higher than any of the other conditions of extraction tested. The duration of the extraction cycle was also evaluated and re-extractions were performed to ensure the full recovery from the samples. It was found that, forcatechin, the R.S.D. of the method is 3.21%, and for epicatechin the R.S.D. was 2.96% (n = 5). The final optimized pressurized liquid extraction method allows for the determination of catechin and epicatechin in diverse types of samples with a rapid (10 min) and reproducible method. PMID- 14763729 TI - Laboratory and field validations of a solid-phase microextraction device for the determination of ethylene oxide. AB - Laboratory and field evaluations were performed to validate a solid-phase microextraction (SPME) device that was used as a diffusive sampler. Hydrogen bromide (HBr) was loaded onto the carboxen-polydimethylsiloxane (CAR-PDMS) fiber for the determination of ethylene oxide (EtO) with on-fiber derivatization. For laboratory evaluations, known concentrations of ethylene oxide around the threshold limit values (TLV)/time-weighted average and specific relative humidities (RHs) were generated by syringe pumps in a dynamic generation system. The SPME diffusive samplers and the commercially available 3M 3551 passive monitors were placed side-by-side in an exposure chamber which was designed to allow measurement of face velocities, temperatures, exposing vapor concentrations, and RHs. Field validations with both SPME diffusive sampler and 3M 3551 passive monitors were also performed. The correlations between the results from both SPME device and 3M 3551 passive monitor were found to be linear (r > 0.9699) and consistent (slope approximately equal to 1.12 +/- 0.07). However, the variations of diffusion coefficients at different temperatures needs to be considered and the adjustment of sampling constant was a must when sampling at temperatures different from 25 degrees C. PMID- 14763730 TI - Determination of free sulfite in wine by zone electrophoresis with isotachophoresis sample pretreatment on a column-coupling chip. AB - This work deals with the determination of free sulfite in wine by zone electrophoresis (ZE) with on-line isotachophoresis (ITP) sample pretreatment on a column-coupling (CC) chip with conductivity detection. A rapid pre-column conversion of sulfite to hydroxymethanesulfonate (HMS), to minimize oxidation losses of the analyte, was included into the developed analytical procedure, while ITP and ZE were responsible for specific analytical tasks in the separations performed on the CC chip. ITP, for example, eliminated the sample matrix from the separation compartment and, at the same time, provided a selective concentration of HMS before its transfer to the ZE stage of the separation. On the other hand, ZE served as a final separation (destacking) method and it was used under the separating conditions favoring a sensitive conductivity detection of HMS. In this way, ITP and ZE cooperatively contributed to a 900 microg/l concentration detectability for sulfite as attained for a 60 nl load of wine (a 15-fold wine dilution and the use of a 0.9 microl sample injection channel of the chip) and, consequently, to the determination of free sulfite when this was present in wine at the concentrations as low as 3 mg/l. The separations were carried out in a closed separation compartment of the chip with suppressed hydrodynamic and electroosmotic flows. Such transport conditions, minimizing fluctuations of the migration velocities of the separated constituents, made a frame for precise migration and quantitation data as achieved for HMS in both the model and wine samples. Ninety percent recoveries, as typically obtained for free sulfite in wine samples, indicate promising potentialities of the present method as far as the accuracies of the provided analytical results are concerned. PMID- 14763731 TI - Screening method for linear alkylbenzene sulfonates in sediments based on water Soxhlet extraction assisted by focused microwaves with on-line preconcentration/derivatization/detection. AB - A screening method for linear alkylbenzene sulfonates (LAS) in sediments has been developed. Soxhlet extraction with water assisted by focused microwaves provides recoveries better (>90%) than obtained by conventional Soxhlet extraction (70 80%). Coupling of the extractor with an on-line preconcentration/derivatization/detection manifold through a flow injection (FI) interface allows a fully automated screening approach. A yes/no answer can be obtained in less than 2 h (for the whole analytical process), a short time compared with the at least 24 h of Soxhlet extraction (without final detection). Due to the use of water as leaching agent, the proposed method is environmentally friendly. PMID- 14763732 TI - Solvent gradient operation of simulated moving beds. 2. Langmuir isotherms. AB - Simulated Moving Bed separations of enantiomers or fine chemicals are usually carried out in the isocratic mode, i.e. by applying the same operating conditions (temperature, pressure, mobile phase composition, pH) in the whole SMB unit. However, it has been recently recognized that by properly modulating operating conditions in the SMB sections. i.e. Sections 1-4 normally, separation performance in terms of productivity and solvent consumption can be significantly improved. In this work, we study solvent gradient SMB (SG-SMB) operation, where the concentration of a modifier in the main solvent constituting the mobile phase is adjusted along the SMB unit, so as to have weaker retention of the species to be separated in the first two sections, and stronger retention in Sections 3 and 4. Overload chromatographic conditions are considered, where the adsorption behavior is characterized by a nonlinear competitive adsorption isotherm, e.g. a binary Langmuir isotherm. Design criteria to achieve complete separation are developed in the frame of the equilibrium theory of chromatography. The theoretical findings are discussed in view of typical effects of the modifier concentration on retention times and solubility of the species to be separated, and an overall assessment of the SG-SMB technology is attempted. PMID- 14763733 TI - High-performance liquid chromatographic separation and quantification of citric, lactic, malic, oxalic and tartaric acids using a post-column photochemical reaction and chemiluminescence detection. AB - An HPLC method was developed for the determination of citric, lactic, malic, oxalic and tartaric acids by chemiluminescent detection following online irradiation with visible light. The organic acids were irradiated with visible light in the presence of Fe3+ and UO2(2+) to generate Fe2+, which was determined by measuring the chemiluminescence intensity in a luminol system in the absence of added oxidant. Factors affecting the photochemical and chemiluminescence reactions were optimised so that their contribution to the total band-broadening was negligible. The chromatographic separation was performed on a C18 column under isocratic reversed-phase conditions using 0.005 M H2SO4 mobile phase. The optimised method was validated with respect to linearity, precision, limits of detection and quantification, accuracy specificity and robustness. The applicability of the assay was demonstrated by analysing these compounds in real samples such as milk, fruit juices, soft drinks, wine and beer. PMID- 14763734 TI - Resolution studies on two regioisomeric chiral stationary phases: effects from reversed orientation of an amide group. AB - Two new polymeric chiral stationary phases, incorporating the selectors trans 9,10-dihydro-9,10-ethanoanthracene-11,12-dicarboxylic acid bis-allylamide, 1 (DEABA) and trans-11,12-diamino-9,10-dihydro-9,10-ethanoanthracene bis butenoylamide, 2 (DDEBB), respectively, have been evaluated by chromatographic resolution of a series of structurally different racemates. For some groups of compounds, where large separation factors were obtained, more detailed studies were performed by the use of different retention modifiers. As an effect from the reversed orientation of the amide group in the two selectors, the enantiomers of the racemates investigated are separated in opposite order of elution on the two columns. PMID- 14763735 TI - Column selectivity in reversed-phase liquid chromatography. V. Higher metal content (type-A) alkyl-silica columns. AB - Retention measurements involving 16 test solutes have been carried out for 38 type-A alkyl-silica columns and three bonded-zirconia columns. These measurements have been analyzed in terms of a model previously developed for type-B columns, so as to yield values of five column selectivity parameters (H, S*, A, B, C) for each type-A column. Overall differences in selectivity between type-A and -B columns can be related to the average values of H, S*, etc. for each column type. Compared to type-B columns, type-A columns provide generally stronger retention for carboxylic acids, while solutes that are more hydrophobic or less bulky are more retained on type-B columns. Hydrogen-bond acceptors (e.g. aliphatic amides) and cations (e.g. protonated bases) are strongly retained on type-A versus type-B columns. Compared to type-B columns, bonded-zirconia columns show much increased retention of cations and reduced retention of hydrogen-bond acceptors. Because of relatively large differences in the selectivity of bonded-zirconia, type-A, and type-B columns, it will prove difficult to find columns of different type (e.g. a type-A and a type-B column) which have equivalent selectivity. Type-A columns also tend to be more different from each other (in terms of selectivity) than is the case for type-B columns. As a result, the replacement of a given type-A column by an "equivalent" type-A column also appears unlikely, except for samples that do not contain ionized compounds. PMID- 14763736 TI - Column selectivity in reversed-phase liquid chromatography. VI. Columns with embedded or end-capping polar groups. AB - A previous model of column selectivity for reversed-phase liquid chromatography (RP-LC) has been applied to an additional 21 columns with embedded or end-capping polar groups (EPGs). Embedded-polar-group columns exhibit a significantly different selectivity vs. non-EPG, type-B columns, generally showing preferential retention of hydrogen-bond donors, as well as decreased retention for hydrogen bond acceptors or ionized bases. EPG-columns are also generally less hydrophobic (more polar) than are non-EPG-columns. Interestingly, columns with polar end capping tend to more closely resemble non-EPG columns, suggesting that the polar group has less effect on column selectivity when used to end-cap the column versus the case of an embedded polar group. Column selectivity data reported here for EPG-columns can be combined with previously reported values for non-EPG columns to provide a database of 154 different columns. This enables a comparison of any two of these columns in terms of selectivity. However, comparisons that involve EPG columns are more approximate. PMID- 14763737 TI - Resolution of a racemic pharmaceutical intermediate. A comparison of preparative HPLC, steady state recycling, and simulated moving bed. AB - Preparative HPLC and simulated moving bed (SMB) chromatography were used to resolve significant quantities of a racemic pharmaceutical intermediate. In addition, smaller scale studies using closed-loop recycling and steady state recycling (SSR) were performed so that a meaningful comparison of all these techniques could be made using the same real world separation. A highly optimized, six-column SMB process was clearly the superior technique and was used for the process-scale (247 kg of racemate) resolution. At the more moderate lab scale (33 kg of racemate and 19 kg of racemate), a frequently used but less optimized eight-column SMB process was used. It was found that SSR was comparable to the lab-scale SMB process in productivity and solvent consumption. Thus, it appears that SSR can be a useful choice at such moderate scales. Finally, at moderate scales when neither SSR nor SMB is available, it was found that acceptable results were obtained with both closed-loop recycling and with a two step preparative process. PMID- 14763738 TI - Dichloropentafluoropropanes as solvents for size exclusion chromatography. AB - We have found that HCFC225s (HCFC225ca: 3,3-dichloro-1,1,1,2,2 pentafluoropropane, HCFC225cb: 1,3-dichloro-1,1,2,2,3-pentafluoropropane) are superior mobile phases for size exclusion chromatography (SEC). As alternatives of CFC113, they have been shown to possess a number of excellent properties, such as low flammability, low viscosity, low cost, high purity, and environmental and operational friendliness. In addition, they have distinct advantages for the SEC measurement, because they solubilize some kinds of acrylate such as poly(methyl methacrylate) (PMMA) and commercial monodispersed PMMA can be used to prepare calibration curves necessary for the measurement of equivalent molecular weight of some polymers. Furthermore, we propose an HCFC225/1,1,1,3,3,3 hexafluoroisopropanol mixed solvent for use in the SEC of poly(ethylene terephthalate) (PET) and polyamides. Poly(2-(perfluorooctyl)ethyl acrylate), whose PMMA equivalent weight average molecular weight was 118,100 Da, was evaluated by a multi-angle laser light scattering (MALLS) detector to have an absolute molecular weight of 439,000 Da. The difference can be attributed to the molecular size of the polyfluorinated polymer compared to the non-fluorinated one. The possible application of this novel mobile phase system for molecular size and molecular weight characterization of perfluoropolyethers, PET, nylon 6 and nylon 6,6 are also discussed. PMID- 14763739 TI - Determining orthogonal and similar chromatographic systems from the injection of mixtures in liquid chromatography-diode array detection and the interpretation of correlation coefficients color maps. AB - Generic orthogonal chromatographic systems might be helpful tools as potential starting points in the development of methods to separate impurities and the active substance in drugs with unknown impurity profiles. The orthogonality of 38 chromatographic systems was evaluated from weighted-average-linkage dendrograms and color maps, both based on the correlation coefficients between the retention factors on the different systems. On each chromatographic system, 68 drug substances were injected as mixtures of three or four components to increase the throughput. The (overlapping) peaks were identified and resolved with a peak purity algorithm, orthogonal projection approach (OPA). The visualization techniques applied allowed a simple evaluation of orthogonal and (groups of) similar systems. PMID- 14763740 TI - Ionic liquid-based liquid-phase microextraction, a new sample enrichment procedure for liquid chromatography. AB - Room temperature ionic liquids (RTILs) were used as extraction solvent in liquid phase microextraction (LPME) coupled with liquid chromatography. Using 1-hexyl-3 methylimidazolium hexafluorophosphate ([C6MIM][PF6]) as extraction solvent, some parameters related to LPME of 4-nonylphenol (4-NP) and 4-tert-octylphenol (4-t OP) were optimized. Although [C6MIM][PF6] can suspend a much larger volume of drop on the needle of the microsyringe than the conventional solvents such as 1 octanol and carbon tetrachloride, the method sensitivity was analyte dependent because of the different partition coefficients and the relatively large viscosity of [C6MIM][PF6]. The proposed procedure has a detection limit and enrichment factor of 0.3 microg l(-1) and 163 for 4-NP, and 0.7 microg l(-1) and 130 for 4-t-OP, respectively. Aqueous samples including tap water, river water, and effluent from sewage treatment plant were analyzed by the proposed method and the recoveries at 10 microg l(-1) spiked level were in the range of 90-113%. PMID- 14763741 TI - Purification of recombinant DNA-derived factor IX produced in transgenic pig milk and fractionation of active and inactive subpopulations. AB - Transgenic animal bioreactors can be engineered to make gram per liter quantities of complex recombinant glycoproteins in milk. However, little is known about the limitations in post-translational processing that occurs for very complex proteins and how this impacts the task of purification. We report on the purification of recombinant factor IX (rFIX) from the milk of transgenic pigs having an expression level of 2-3 g rFIX/(l(-1) h(-1)), an expression level that is about 20-fold higher than previously reported. This purification process efficiently recovers highly active rFIX and shows that even complex mixtures like pig milk, which contains 60 g/l total endogenous milk protein and multiple subpopulations of rFIX, can be processed using conventional, non-immunoaffinity chromatographic methods. Without prior removal of caseins, heparin-affinity chromatography was used to first purify the total population of rFIX at greater than 90% yield. After the total population was isolated, the biologically active and inactive subpopulations were fractionated by high-resolution anion exchange chromatography using an ammonium acetate elution. Capillary isoelectric focusing of the active and inactive rFIX fractions demonstrated that the active subpopulations are the most acidic. PMID- 14763742 TI - Identification and characterization of Se-methyl selenomethionine in Brassica juncea roots. AB - The present work shows the identification and characterization of Se-methyl selenomethionine (SeMMet) as an important Se species in Brassica juncea roots when grown in the presence of Se-methionine (SeMet) as the Se source. SeMMet was isolated by liquid chromatography employing two different liquid chromatographic mechanisms: reversed-phase ion-pairing using heptafluorobutyric acid as counter ion and cation exchange using a pyridinium formate gradient (pH 3). Inductively coupled plasma mass spectrometry was used for the detection of Se. SeMMet was characterized by electrospray quadrupole time-of-flight MS in both a synthesized standard and in the roots extract using collision-induced dissociation of the selected ion. Preliminary evidence suggests that Brassica juncea may also produce dimethylselenonium propionate, although to a much lesser extent. PMID- 14763743 TI - Ion-chromatographic determination of carbocisteine in pharmaceuticals based on non-suppressed conductimetric detection. AB - A novel method for the determination of carbocisteine (S-CMC), a mucolytic and expectorant drug with an acidic amino acid structure, was developed and validated, using non-suppressed ion-chromatographic system with conductimetric detection, and anion or cation exchange columns. Among the various combinations of column type and eluent composition tested, a cation exchange column with a 0.25 mM tri-fluoroacetic acid (TFA) as eluent in isocratic mode at 1.2 ml/min gave the best results. S-CMC was very well separated from all common amino acids (resolution > 2.6). The retention time was 3.5 min and the asymmetry factor 1.1. A linear calibration curve from 17 to 400 microg/ml (r = 0.99994), with a detection limit of 0.14 microg (5.6 microg/ml-25 microl injection volume) and a precision of 1.5% R.S.D. (100 microg/ml, n = 3) was achieved. The proposed method was applied for the determination of S-CMC content in intensely colored commercial formulations (syrups). No interference from excipients was found and the only pretreatment step was the appropriate dilution with the mobile phase. Recovery from standard additions was ranged from 96.0 to 104.9% and precision (R.S.D., n = 3) 1.8-3.6%. PMID- 14763744 TI - Quantification of risperidone and 9-hydroxyrisperidone in plasma and saliva from adult and pediatric patients by liquid chromatography-mass spectrometry. AB - A robust and validated LC-MS-MS quantitative method, using column switching and mutiple reaction monitoring was developed for the analysis of risperidone (RIS) and 9-hydroxyrisperidone in human plasma and saliva. The analytical range was 1 100 ng/ml. The method used 25 microl of sample precipitated with 75 microl of acetonitrile containing internal standard (R068808). Analyses were conducted on a PE Sciex API-III + triple quadrupole mass spectrometer fitted with a Turbo IonSpray source. The method was validated for human plasma using EDTA as the anticoagulant and cross-validated to heparinized human plasma and saliva. The recoveries of risperidone and 9-hydroxyrisperidone were 90-93 and 89-93%, respectively. The validated method was applied to clinical samples to study risperidone and 9-hydroxyrisperidone concentrations in plasma and saliva. Risperidone and 9-hydroxyrisperidone appear in the saliva of patients treated with risperidone. Their detection/quantification in saliva provides evidence for recent adherence with therapy. PMID- 14763745 TI - Application of preparative high-speed counter-current chromatography for separation of chlorogenic acid from Flos Lonicerae. AB - Chlorogenic acid, an ester formed between caffeic acid and quinic acid, is a major phenolic compound in the traditional Chinese medicinal herb Flos Lonicerae. The separation and purification of chlorogenic acid from the crude extract of Flos Lonicerae was achieved by high-speed counter-current chromatography (HSCCC). A high acid, highly polar two-phase solvent system containing n-butanol-acetic acid-water (4:1:5) was run on a preparative scale. The upper phase was used as the mobile phase in the head to tail elution mode. A 300-mg quantity of the crude extract containing 5.97% chlorogenic acid was loaded on a 342-ml HSCCC column. Double separations were performed with the same solvent system yielding 16.9 mg chlorogenic acid at 94.8% purity with approximately 90% recovery. PMID- 14763746 TI - Dodecylsulfate-coated monolithic octadecyl-bonded silica stationary phase for high-speed separation of hydrogen, magnesium and calcium in rainwater. AB - The high-speed determination of hydrogen, magnesium and calcium ions by ion chromatography (IC) is demonstrated on a monolithic octadecyl-boned silica (ODS) column coated with lithium dodecylsulfate (Li-DS). This stationary phase, when used in conjunction with a 2 mM ethylenediamine and 0.1 mM Li-DS solution as eluent at pH 6.0, was found to be suitable for the rapid and efficient separation of hydrogen and magnesium and calcium in the order H+ < Mg2+ < Ca2+ within 4 min at a flow rate of 4.0 ml/min. Under the conditions, linear calibration plots of conductivity versus concentration were obtained for the cations over about three orders of magnitude, and the detection limits were 1 microM for H+, 2 microM for Mg2+ and Ca2+. Rainwater was analyzed directly using this IC system with satisfactory results. PMID- 14763747 TI - Coupled ion chromatography for the determination of chloride, phosphate and sulphate in concentrated nitric acid. AB - A coupled ion chromatography (IC) system was used for the determination of chloride, sulphate and phosphate in high-purity nitric acid. Such a high ionic strength matrix causes a selectivity problem in single IC systems. The first part of the system is used for a pre-separation of the analytes from the nitrate matrix. A specially designed high-capacity anion exchanger with low retention for the analytes and high retention for nitrate was developed. The eluent stream containing the analytes was transferred to the second part of the system via a heart-cut valve and a pre-concentration column. The second system utilizes a high performance anion exchanger and is used to quantify the analytes. Recoveries of the analytes are 80-100% for phosphate, and around 100% for sulphate and chloride. Detection limits for chloride, sulphate and phosphate in concentrated nitric acid (69% w/w) are 0.1, 1 and 5 mg/l, respectively. PMID- 14763748 TI - Prediction of the resolution of capillary columns in different conditions of inlet pressure and temperature. AB - A procedure previously described for the prediction of the plate height of capillary columns operated at different inlet pressure of the carrier gas and at various column temperatures by using few retention data measured under isobaric conditions was modified and improved in order to permit the prediction of the retention times and of the peak widths at various heights. It is therefore possible to calculate the ratio, delta, between the peak width at different heights and the peak width at half height, whose value is used to predict the resolution at different height of two closely eluting peaks. It was found that the delta values do not depend on temperature and inlet pressure and are a characteristic of the used column; they can therefore be used in order to calculate the resolution in any temperature and inlet pressure condition. The method was used to predict the retention time, the peak width and the resolution of polar and non-polar compounds (alkanes, alkenes, chloroalkanes, alcohols, ketones) on capillary columns of different length and polarity by using as the starting data retention and width values measured in three isobaric runs only. PMID- 14763749 TI - Analysis of polycyclic aromatic hydrocarbons in contaminated soil by Curie point pyrolysis coupled to gas chromatography-mass spectrometry, an alternative to conventional methods. AB - Curie point pyrolysis gas chromatography-mass spectrometry (Py-GC-MS) has been compared with classical extraction procedures (Soxhlet, sonication, KOH digestion, microwave-assisted) followed by GC-MS analysis for the determination of polycyclic aromatic hydrocarbons (PAHs) in contaminated soil. In each case, the efficiency of the technique was examined for 16 PAHs included in the US Environmental Protection Agency Priority Pollutant List. The results indicate that the recovery of PAHs is dependent on the extraction technique. The highest recoveries of PAHs were obtained with Curie point pyrolysis and KOH digestion. Py GC-MS appeared to be interesting alternative method for the determination of PAHs in contaminated soil. The results were validated by certified soil (CRM 104) analysis. PMID- 14763750 TI - Monitoring coal-tar pitch composition changes during air-blowing by gas chromatography. AB - A series of air-blown coal-tar pitches was studied by GC and GC-MS in order to achieve a deeper understanding of the behaviour of the different pitch components during air-blowing. Compounds present in the parent pitch were identified and quantified and then compared to those present in the air-blown pitches. The compounds observed were identical before and after the treatments, but the concentration of each compound changed with the treatment to a different extent depending on its molecular structure and consequently its reactivity to oxygen. The most reactive compounds were those with a mehylene-bridge in a five member ring, followed by those with a methyl group in their structure. PMID- 14763751 TI - Solid-phase micro-extraction-gas chromatography-(tandem) mass spectrometry as a tool for pesticide residue analysis in water samples at high sensitivity and selectivity with confirmation capabilities. AB - Gas chromatography-mass spectrometry (GC-MS) has been widely applied for pesticide monitoring because of its high sensitivity and specificity and for the potential of multi-residue and multi-class analysis. An analytical procedure was developed for the determination of pesticide multi-residues in water samples combining solid-phase micro-extraction (SPME) and gas chromatography-ion trap mass spectrometry. For SPME extraction a poly(dimethylsiloxane)-divinylbenzene coated fibre was selected whereas the mass spectrometer was operated under full scan, selected ion storage (SIS), microSIS (SIM) and MS-MS and the figures of merit compared. Quantitative and qualitative (confirmatory) capabilities of each operation mode are discussed. Using MS-MS, precision was typically below 10% and limits of detection (LODs) were improved by 1.3 to 20 times (to low- or sub-ppt levels) compared to microSIS, with the advantage of maintaining identification capabilities. The combination of selective extraction by SPME and highly selective determination by GC-MS-MS made possible ultra-selective and essentially error-free determination of pesticides in complex environmental samples. This aspect will be highlighted in the paper. PMID- 14763752 TI - Determination of ionisation constants of organic bases in aqueous methanol solutions using capillary electrophoresis. AB - The pKa of eight organic bases was determined in aqueous and aqueous methanol solutions of 0-70% (v/v) methanol using capillary electrophoresis. The bases investigated include compounds commonly used to test the activity of RP columns in HPLC. The variation of pKa with temperature in aqueous methanol solutions was also investigated and found to closely resemble temperature coefficients reported for bases in purely aqueous solutions. pKa values determined by CE were compared to those reported using NMR spectroscopy. The good agreement of the results is evidence that either technique is suitable to perform pKa measurements. PMID- 14763753 TI - Speciation of inorganic and methylated arsenic compounds by capillary zone electrophoresis with indirect UV detection. Application to the analysis of alkali extracts of As2S2 (realgar) and As2S3 (orpiment). AB - A capillary zone electrophoresis (CZE) method with indirect UV detection was developed to simultaneously separate inorganic and organic arsenic compounds including arsenite (iAsIII), arsenate (iAsV), monomethylarsonate and dimethylarsenic acid (DMAV). 2,6-Pyridinedicarboxylic acid (PDC) and n hexadecyltrimethylammonium hydroxide (CTAOH) were selected to compose a background electrolyte (BGE), where PDC was used as chromophore and CTAOH functioned as electroosmotic flow (EOF) modifier to reduce/eliminate EOF. The choice of detection wavelength, the optimization of BGE pH, and effects of applied electric field strength and temperature on separation were further investigated. The limits of detection for the targeted analytes were between 0.19 and 0.23 ppm as molecule. Good linearity of more than three orders of magnitude was obtained. Repeatability of migration times and peaks areas were 0.8-1.7 and 3.4-6.9% R.S.D.; whereas reproducibility were 1.2-2.2 and 3.6-7.1% R.S.D., respectively. The established CZE method was then applied to analyze the alkali extracts of realgar (As2S2) and orpiment (As2S3). The main components in both alkali extracts were identified to be iAsIII and iAsV. PMID- 14763754 TI - Determination of average degree of polymerisation and distribution of oligosaccharides in a partially acid-hydrolysed homopolysaccharide: a comparison of four experimental methods applied to mannuronan. AB - The average degree of polymerisation (DP) and distribution of oligosaccharides in partially acid hydrolysed mannuronans were quantitatively evaluated by 1H NMR, electrospray ionisation mass spectrometry (ESI-MS), micellar electrokinetic capillary chromatography with UV detection (MEKC-UV), and high-pressure anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD). Our investigation shows that 1H NMR, MEKC-UV and, in particular, HPAEC-PAD can be used as quantitative tools to aid the investigation of polysaccharide structure, function and synthesis. For the latter two techniques, especially, this represents a significant new development as it enables calculation of the quantity of individual oligomers of nominal DP by direct analysis of a defined oligomer mixture. Appropriate statistical averages of number and weight distributions were also calculated and found to fit very well to predicted Kuhn distributions that assume random depolymerisation. PMID- 14763755 TI - Evaluation of multi-walled carbon nanotubes as an adsorbent for trapping volatile organic compounds from environmental samples. AB - A type of purified multi-walled carbon nanotubes (PMWCNTs) prepared by catalytic decomposition of methane, with a surface area of 98 m2/g, was evaluated as an adsorbent used for tapping volatile organic compounds (VOCs). The performance in evaluation was based on breakthrough volumes (BTVs) and recoveries of selected VOCs. PMWCNTs were also used as a trap packing material to adsorb VOCs purged from spiked water sample. Due to their porous structure, PMWCNTs were found to have much higher BTVs than that of Carbopack B, a graphitized carbon black with the same surface area as PMWCNTs. The recoveries of the tested VOCs trapped on PMWCNTs ranged from 80 to 110%, and not affected by the humidity of purge gas. The results indicate that PMWCNTs are a potential useful adsorbent for direct trapping VOCs from air samples and may be a supplement to VOCARB 3000, a commercially available trap, in purge-and-trap system to preconcentrate VOCs from water samples. PMID- 14763756 TI - Determination of residues of UV filters in natural waters by solid-phase extraction coupled to liquid chromatography-photodiode array detection and gas chromatography-mass spectrometry. AB - This study describes a procedure for the enrichment, separation and quantification of four major UV filters in natural waters. Solid-phase extraction combined with liquid chromatography and photodiode array detection (LC-UV-DAD), and gas chromatography with mass spectroscopy (GC-MS) were employed for the analyses. LC of the four compounds with surfactant-modified hydro-organic eluents gave satisfactory resolution of overlapping peaks. In GC, a significant improvement of the detection limits was attained, but only three compounds could be detected. The method was validated for, and applied to, various water samples prone to UV filter accumulation due to recreational activities. Recoveries from real samples were 86-99% with LOQs as low as 0.5 ng/l depending on the sample volume and the analytical procedure. PMID- 14763757 TI - Suitability of N,O-bis(trimethylsilyl)trifluoroacetamide and N-(tert butyldimethylsilyl)-N-methyltrifluoroacetamide as derivatization reagents for the determination of the estrogens estrone and 17alpha-ethinylestradiol by gas chromatography-mass spectrometry. AB - This paper describes apreviously unreported problem with the use of N,O-bis (trimethylsilyl)trifluoroacetamide (BSTFA) and N-(tert-butyldimethylsilyl)-N methyltrifluoroacetamide (MTBSTFA) to derivatise the natural hormone estrone (E1) and the synthetic estrogen 17alpha-ethinylestradiol (EE2). The resulting trimethylsilyl (TMS) and t-butyldimethylsilyl (TBS) derivatives of EE2 were partially converted to their respective El derivatives. Therefore, these reagents may not be suitable for simultaneous determination of estrogens in environmental samples, which raises questions about the reliability of results from some earlier studies. PMID- 14763758 TI - Simplified and rapid method for extraction of ergosterol from natural samples and detection with quantitative and semi-quantitative methods using thin-layer chromatography. AB - A new and simplified method for extraction of ergosterol (ergosta-5,7,22-trien 3beta-ol) from fungi in soil and litter was developed using pre-soaking extraction and paraffin oil for recovery. Recoveries of ergosterol were in the range of 94-100% depending on the solvent to oil ratio. Extraction efficiencies equal to heat-assisted extraction treatments were obtained with pre-soaking extraction. Ergosterol was detected with thin-layer chromatography (TLC) using fluorodensitometry with a quantification limit of 8 ng. Using visual evaluation of images of TLC plates photographed in UV-light the quantification limit was 16 ng. PMID- 14763759 TI - Contrast sensitivity loss with aging: sampling efficiency and equivalent noise at different spatial frequencies. AB - The relative contributions of optical and neural factors to the decrease in visual function with aging were investigated by measurement of contrast detection at three different spatial frequencies, in the presence of external noise, on young and older subjects. Contrast detection in noise functions allows two parameters to be measured: sampling efficiency, which indicates neural changes, and equivalent noise, which demonstrates optical effects. Contrast thresholds were measured in the presence of four levels (including zero) of externally added visual noise. Measurements were obtained from eight young and eight older visually normal observers. Compared with young subjects, older subjects showed significantly (p < 0.05) lower sampling efficiencies at spatial frequencies of 1 and 4 cycles per degree (c/deg) and significantly higher equivalent noise levels for gratings of 10 c/deg. Neural and optical factors affect contrast sensitivity loss with aging differently, depending on the spatial frequency tested, implying the existence of different mechanisms. PMID- 14763760 TI - Corresponding-pair procedure: a new approach to simulation of dichromatic color perception. AB - The dichromatic color appearance of a chromatic stimulus T can be described if a stimulus S is found that verifies that a normal observer experiences the same sensation viewing S as a dichromat viewing T. If dichromatic and normal versions of the same color vision model are available, S can be computed by applying the inverse of the normal model to the descriptors of T obtained with the dichromatic model. We give analytical form to this algorithm, which we call the corresponding pair procedure. The analytical form highlights the requisites that a color vision model must verify for this procedure to be used. To show the capabilities of the method, we apply the algorithm to different color vision models that verify such requisites. This algorithm avoids the need to introduce empirical information alien to the color model used, as was the case with previous methods. The relative simplicity of the procedure and its generality makes the prediction of dichromatic color appearance an additional test of the validity of color vision models. PMID- 14763761 TI - Influence of imperfections in gradient-index waveguides on Talbot effects. AB - On the basis of the unitary transformation and the Lie algebra decomposition technology widely used in quantum mechanics, we obtain the analytical propagator for light beams propagating through an imperfect gradient-index (GRIN) waveguide. The results show that, unlike in the straight GRIN waveguide widely studied, in an imperfect GRIN waveguide, self-imagining phenomena can result in two new effects: one is a phase shift including a global one and a local one, in which the local one results in a change of direction of the light beam propagating through the imperfect GRIN waveguide; the other is a transverse shift of self image. The transverse deviation occurring in the imperfect GRIN waveguide is also calculated. PMID- 14763762 TI - Parametric characterization of rotationally symmetric hard-edged diffracted beams. AB - The truncated second-order moments and generalized M2 factor (M(G)2 factor) of two-dimensional beams in the Cartesian coordinate system are extended to the case of three-dimensional rotationally symmetric hard-edged diffracted beams in the cylindrical coordinate system. It is shown that the propagation equations of truncated second-order moments and the M(G)2 factor take forms similar to those for the nontruncated case. The closed-form expression for the M(G)2 factor of rotationally symmetric hard-edged diffracted flattened Gaussian beams is derived that depends on the truncation parameter beta and beam order N. For N --> infinity, the M(G)2 factor equals 4/square root of 3 corresponding to the value of truncated plane waves, which guarantees consistency of the formalism. PMID- 14763763 TI - Representing the behavior of partially coherent optical systems by using overcomplete basis sets. AB - A technique is described for representing the behavior of partially coherent optical systems by using overcomplete basis sets. The scheme is closely related to Gabor function theory. Through singular-value decomposition it is shown that if E is a matrix containing the sampled basis functions, then all of the information needed for optical calculations is contained in S = EE(dagger) and R = E(dagger)E. For overcomplete sets, S can be inverted to give a dual basis set, E = S(-1)E, which can be used to find the correlation matrix elements A of a sampled bimodal expansion of the spatial coherence function. Overcomplete correlation matrices can be scattered easily at optical components. They can be used to determine (i) the natural modes of a field; (ii) the total power in a field, Pt = Tr[RA]; (iii) the power coupled between two fields, A and B, that are in different states of coherence, Pc = Tr[RARB]; and (iv) the entropy of a field, Q = Tr[Zsigmar(I-Z)r/r], where Z = RA/Tr[RA]. PMID- 14763764 TI - Polarimetric description of superposing random electromagnetic fields with different spectral composition. AB - We present a polarimetric technique that provides a complete description of a mixture of uncorrelated optical fields with different spectral and polarization properties. The second-order coherence theory is used to describe the superposition of two random optical fields, and an imaging experiment is reported that illustrates the capability to separate radiations with different spectral composition and to simultaneously determine their Stokes vectors. PMID- 14763765 TI - Time-sharing wave-front-sensing adaptive optics. AB - Based on the concept of common-path/common-mode adaptive optics, the time-sharing wave-front-sensing adaptive optics system contains only one Hartmann-Shack (H-S) wave-front sensor, which detects two aberrations in the beam path alternately. After data fusion of the two aberrations, the actuator voltage of the deformable mirror (DM) is obtained. How the disturbances of the slope data and the response matrix influence the DM's actuator voltage in the data fusion methods is discussed, and the effective upper limits are given. Feasible data fusion methods are tested, and experiments verify that the performance of the system is good. The time-sharing technique is limited in sampling rate and is suitable only for corrections of slowly changing phases, because the H-S wave-front sensor's sampling frequency must be adequate for the alternate detection of two aberrations. PMID- 14763766 TI - Singular-value analysis and optimization of experimental parameters in fluorescence molecular tomography. AB - The advent of specific molecular markers and probes employing optical reporters has encouraged the application of in vivo diffuse tomographic imaging at greater spatial resolutions and hence data-set volumes. This study applied singular-value analysis (SVA) of the fluorescence tomographic problem to determine optimal source and detector distributions that result in data sets that are balanced between information content and size. Weight matrices describing the tomographic forward problem were constructed for a range of source and detector distributions and fields of view and were decomposed into their associated singular values. These singular-value spectra were then compared so that we could observe the effects of each parameter on imaging performance. The findings of the SVA were then confirmed by examining reconstructions of simulated and experimental data acquired with the same optode distributions as examined by SVA. It was seen that for a 20-mm target width, which is relevant to the small-animal imaging situation, the source and detector fields of view should be set at approximately 30 mm. Equal numbers of sources and detectors result in the best imaging performance in the parallel-plate geometry and should be employed when logistically feasible. These data provide guidelines for the design of small animal diffuse optical tomographic imaging systems and demonstrate the utility of SVA as a simple and efficient means of optimizing experimental parameters in problems for which a forward model of the data collection process is available. PMID- 14763767 TI - Analysis of low-coherence interference fringes by the Kalman filtering method. AB - Interferometers with low-coherence illumination allow noncontact measurement of rough-surface relief with a wide range of measurement definition by locating the visibility maxima of interference fringes. The problem is light scattering by the surface to be measured, which can cause distortion of low-coherence interferometric signals. We propose to use a stochastic fringe model and a Kalman filtering method for processing noisy low-coherence fringes dynamically. Prediction of the fringe's signal value at each discretization step is based on all the information available before this step; the prediction error is used for dynamic correction of the estimates of the fringe envelope and phase. The advantages of the Kalman filtering method consist in its immunity to noise, optimal fringe evaluation, and data-processing speed. PMID- 14763768 TI - Measurement of random processes at rough surfaces with digital speckle correlation. AB - We present a detailed investigation of digital speckle correlation to measure small changes in the microstructure of random rough surfaces. The corresponding alterations in the scattered-light field are recorded by an electronic camera with subsequent numerical correlation. Among the classical theoretical approaches to describe the scattering at random rough surfaces, the composite-roughness model is advanced to calculate the speckle correlation in terms of parameters of the changes in surface microstructure. For an experimental verification, surfaces with similar microstructure are fabricated with a photolithographic technique. They are employed for comparative measurements with high-resolution scanning force microscopy and for correlation measurements under variation of experimental parameters. A good agreement between theoretically predicted and experimental correlation data can be observed. The results allow a quantitative whole-field monitoring of surface processes by remote optical means. PMID- 14763769 TI - Efficient numerical method for predicting the polarization-dependent Raman gain in fiber Raman amplifiers. AB - Polarization-dependent gain (PDG) of fiber Raman amplifiers (FRAs) will degrade the performance of optical communication systems. An efficient numerical model is presented to predict PDG quantitatively by substituting the polarization dependent polarization factor for the constant one in the coupled nonlinear equations usually adopted. The simulation is carried out by estimating the polarization length by use of the average polarization-mode dispersion of the tested fiber; the results, including the Raman gain profile and the fluctuation of the PDG, are highly accordant with the experimental data reported previously. The model can aid in the design of FRAs and in the analysis of system performance. PMID- 14763770 TI - Analysis of coupling between two slab waveguides in the presence of ring resonators. AB - The coupling phenomena between two slab waveguides in the presence of ring resonators are investigated through a rigorous integral equation analysis. A Green's-function-theory approach is utilized to develop the integral equation formulation. The solution is obtained by applying an entire-domain Galerkin technique, using the orthogonality properties of involved wave functions in the ring resonators. The field's transmission and reflection coefficients are accurately computed and a practical directional-coupler approach is discussed. The presented results reveal strong resonant coupling phenomena between the two waveguides and the resonators. PMID- 14763771 TI - Theory for the measurements of dispersion characteristics in waveguiding structures with a scanning near-field optical microscope. AB - A theory is presented for the interpretation of scanning near-field optical microscope measurements on pulses propagating in waveguiding structures. It is shown how the dispersion characteristics of the propagating guided modes may be derived from such experiments. Then it is demonstrated how to calibrate the scanning tip position and to derive experimental values for reflection and transmission of modes in identical single-mode waveguides connected to a photonic device such as a micro cavity. PMID- 14763772 TI - Shape evolution of a light pulse in a linear birefringent medium. AB - The spreading of a three-dimensional quasi-monochromatic progressive directional wave packet (such as a laser pulse) propagating freely in a linear and transparent birefringent medium is described geometrically by means of the ellipsoid representative of the pulse's second-order moments. The medium is characterized by a second-order expansion of its dispersion relation omega(K) about the mean wave vector Km of the pulse, i.e., by its Hessian matrix (H(Km)omega), which plays two important roles. Then, for some elements of(H(Km)omega), practical expressions are provided that are related to the curvature and dispersion properties of the normal surface of the medium. Then cases in which these properties determine completely the asymptotic transverse or axial spreadings of the wave packet are specified, and the results of an experiment are discussed. PMID- 14763773 TI - Efficient method for the determination of extreme-ultraviolet optical constants in reactive materials: application to scandium and titanium. AB - The chemical reaction of a sample with atmospheric gases causes a significant error in the determinantion of the complex refractive index n = 1 - delta + ibeta in the extreme-ultraviolet region. The protection of samples removes this effect but hampers the interpretation of measurements. To overcome this difficulty, we derive the exact dependences on film thickness of the reflectivity and transmissivity of a protected film. These dependences greatly simplify the determination of delta and beta when the spectra of several films with different thickness and identical protection are measured. They also allow the verification of the delta(omega) obtained from the Kramers-Kronig relation and even make the Kramers-Kronig method unnecessary in many cases. As a practical application, the optical constants of Sc and Ti are determined at h omega = 18-70 eV and 18-99 eV, respectively. The essential feature of our experimental technique is deposition of a film sample directly on a silicon photodiode that allows easy operation with both thin (approximately 10-nm) and thick (approximately 100-nm) films. The comparison of calculated reflectivities of Si-Sc multilayers with the measured values shows the high accuracy of the determined delta(omega) and beta(omega). PMID- 14763774 TI - Museum lighting: why are some illuminants preferred? AB - We had shown earlier that viewers prefer to look at artworks under illuminants of approximately 3600 K. In the latest paper we tested the hypothesis that the preferred illuminant is one that appears neither warm nor cool and repeated the settings at each of four illuminances to test the stability of the findings. Observers looked at a neutral white reflectance standard hung on a matte-gray wall lit by overhead banks of lamps whose combined value could be adjusted continuously between 3000 and 4400 K while illuminance was kept constant. Illuminance ranged from 50 to 2000 lux. Observers adjusted color temperature until they were satisfied that the standard looked neither warm nor cool. The mean for a group of eight observers was approximately 3700, independent of intensity; this corresponds to a dominant wavelength of approximately 580 nm. In a separate study four observers scaled the apparent warmth or coolness of flashes of equiluminant monochromatic lights; the warm-cool transition was between 560 and 580 nm; warmness was completely predicted by the perceived redness of each light as derived from hue and saturation scaling functions from the same group. PMID- 14763775 TI - Endodontic variables and coronary heart disease. AB - This cross-sectional study was designed to explore a possible association between endodontic disease variables and coronary heart disease (CHD). Dental infections are hypothesized to be linked to atherosclerosis and could be a cause of vascular changes crucial for the development of CHD. Most studies have focused on periodontal disease. To our knowledge, no one has specifically studied endodontic variables as risk factors for the development of CHD. In 1992-93, a representative sample (n = 1056) of women in Goteborg, Sweden, aged between 38 and 84 years, took part in a combined dental and medical survey. The dependent variable was CHD, i.e. subjects with angina pectoris and/or a history of myocardial infarction (n = 106). The independent variables were number of root filled teeth (RF), number of teeth with periapical radiolucencies (PA), tooth loss (TL), age, life situation, marital status, smoking, alcohol habits, body mass index, waist-hip ratio, serum cholesterol and triglyceride concentrations, hypertension and diabetes. The multivariate logistic regression analysis did not prove the endodontic variables to be predictive of CHD. Only age and tooth loss were significantly associated with CHD, with OR = 1.07 (CI = 1.03-1.12) and OR = 2.70 (CI = 1.49-4.87), respectively. The bivariate logistic regression analysis showed a positive significant association between subjects with RF = 2 and CHD, but for PA the bivariate analysis did not support an association with CHD. This cross-sectional study did not reveal a significant association between endodontically treated teeth and CHD nor between teeth with periapical disease and CHD. PMID- 14763776 TI - Radiographic findings on 3rd molars removed in 20-year-old men. AB - In this study we assess radiographic findings characteristic of mandibular 3rd molars that had required either routine or surgical extraction. X-ray findings relating to acute pericoronitis were also examined. The material was collected by investigating patient records and rotational panoramic radiographs of 20-year-old Finnish male conscripts (n = 738) treated during military service because of 3rd molar-related problems. The follicle around the crown of mandibular 3rd molars with acute pericoronitis was enlarged in 19% of cases and in 13% of chronic symptom-free pericoronitis cases (not statistically significant difference). Mandibular 3rd molars extracted surgically were more often mesially inclined than those extracted routinely (61% vs. 23%; P < 0.001), partially or totally intrabony impacted (92% vs. 66%; P < 0.001) and deep situated (on average 4.2 mm vs. 2.5 mm under the occlusal plane). Surgical extraction was also associated with the roots completely developed [92% vs. 84% of the teeth routinely extracted, odds ratio (OR) 2.6, 95% confidence interval (CI) 1.2-5.5] and with the absence of radiographic pericoronitis [around 27% vs. 39% of the teeth routinely extracted (OR 0.5, 95% CI 0.3-0.8)]. In 86% of cases the space between 2nd molar and ramus of the mandible was narrower than the 3rd molar extracted surgically, whereas this was 62% in routine extraction cases (P < 0.001). We conclude that there are some typical 3rd-molar findings in rotational panoramic radiographs that show a need for surgical extraction. PMID- 14763777 TI - Intra-oral adhesive systems for ceramic repairs: a comparison. AB - The aim of this investigation was to compare the bond strength of restorative composite resin to dental ceramic conditioned with primers and adhesives of various commercial repair kits. Three intra-oral ceramic repair systems--Silistor (Heraeus Kulzer), Cimara (Voco), Ceramic Repair (Vivadent)--were used on all ceramic (IPS Empress 2, Ivoclar-Vivadent) substrate. Shear bond strength of restorative composite resin to substrate was tested after thermocycling and without thermocycling (n = 10). Substrate surfaces of the specimen after loading were examined microscopically (SEM). The highest bond strengths in both water stored (7.0 +/- 5.7 MPa) and thermocycled conditions (2.5 +/- 1.8 MPa) were obtained with the Vivadent repair system, while the lowest values were observed with the Cimara system (0.6 +/- 1.4 MPa and 0.0 +/- 0.0 MPa, respectively). Shear bond strengths appeared to be significantly affected by thermocycling (ANOVA, P < 0.05). It is concluded that there are significant differences in the bond strengths of resin composites and ceramic substrate. The roughened surface does not necessarily provide a better bond strength; the bond strength of composite decreases with storage in water and after thermocycling. Bond strength values were generally low for all of the tested materials. PMID- 14763778 TI - Effect of variable moderate chronic stress on ligature-induced periodontal disease in Wistar rats. AB - The aim of this investigation was to study the impact of stress on ligature induced periodontal disease in rats by means of a variable moderate chronic stress model. Thirty male Wistar rats were randomly assigned to six groups. Control groups received only ligatures around the second maxillary molars, while experimental groups were exposed to stress in additional. Stress was imposed by means of flashing light, isolation, rat blood smelling, new environment exposure, immobilization in cold temperature and immobilization at room temperature. Stress was applied randomly, thereby diminishing adaptation of the animals to the model. The animals were killed after 29, 43, and 57 days. The distance between the cementum-enamel junction and the alveolar bone (CEJ-AB) was measured. Alveolar bone loss was statistically different between stressed and control animals, whereas differences were not observed between experimental periods. The mean CEJ AB distance in animals exposed to stress was 154.50 microm smaller than the corresponding distance in the controls. It might be concluded that variable moderate chronic stress decreased alveolar bone loss in a ligature-induced periodontal disease model. PMID- 14763779 TI - Impact of temporomandibular joint pain on activities of daily living in patients with rheumatoid arthritis. AB - The aim of this study was to investigate the impact of temporomandibular joint (TMJ) pain on daily living in patients with rheumatoid arthritis (RA) involving the TMJ. Nineteen patients (17 F, 2 M) with a median (IQR) age of 44 (23) years were included. A scale for the influence of TMJ pain/discomfort on the activities of daily living was used. TMJ resting pain and pain upon maximum mouth opening according to a visual analog scale as well as pressure pain threshold and tenderness to digital palpation of the TMJ were assessed. Blood samples were collected to measure the level of acute phase proteins. Activities of daily living were influenced in all patients at different levels. The impact on daily living by TMJ pain/discomfort was greatest on the performance of physical exercises and jaw movements, while it was smallest on the performance of hobbies and eating. Pain during maximum mouth opening and tenderness to digital palpation were correlated to difficulties with several activities such as to yawn and open the mouth wide, while pressure pain threshold was correlated with difficulties during eating, which confirms that the pain was located in the TMJ. In conclusion, this study indicates that pain/discomfort from the TMJ in patients with RA has a significant negative impact on activities of daily living. PMID- 14763780 TI - Dental anxiety in Iceland: an epidemiological postal survey. AB - In this study, we examined the prevalence of specific (dental) phobia among a sample of the Icelandic population. In addition to dental anxiety we explored factors that could be related to dental anxiety. In the period 1972-73, a stratified sample of 1641 schoolchildren in Reykjavik was selected for a study on malocclusion, dental maturation and other factors. Twenty-two years later (1995), a postal survey conducted in this group looked at many variables relating to oral health, including orofacial pain, functional oral disorders, self-perception of dental and general appearance and need for orthodontic treatment. Out of 1529 individuals contacted, 1192 completed questionnaires were returned (response rate 78%). Questions based on DSM-IV criteria of specific (dental) phobia (DP) were included. Ninety-six participants reported that they had avoided dental treatment during the previous 6 months. Twenty-one respondents fulfilled DSM-IV criteria for specific (dental) phobia (DP) and 75 admitted to many symptoms of dental anxiety (DA). Specific (dental) phobia (DP) was more prevalent among women than among men. The divorced or widowed were most at risk, as were non-salaried respondents. Most respondents attributed the onset of their phobias to a specific painful or fearful experience. There was a significant difference between the total dentally anxious (TDA = DA + DP) and the not dentally anxious (NDA) with regard to sex (women--higher TDA) and marital status (divorced or widowed--higher TDA). The TDA had statistically fewer teeth than the NDA and received dental treatment less frequently. PMID- 14763781 TI - Fluoride release from NaF- and AmF-impregnated toothpicks and dental flosses in vitro and in vivo. AB - The aim of this study was to determine the fluoride release from toothpicks and dental flosses in vitro and in vivo, and to evaluate various approximal administration methods. In vitro, a total of 23 commercially available toothpicks and dental flosses and 3 prototypes impregnated with sodium fluoride (NaF), amine fluoride (AmF) or a combination of these two were tested. Fluoride release was determined for up to 24 h using an ion-specific electrode. A large variation was found between the products; most fluoride being released after 30 min. Generally speaking, toothpicks produced higher values than flosses. In vivo, the fluoride concentration in both treated and non-treated approximal areas was evaluated after using 2 different types of toothpicks and 4 dental flosses and after different application methods--such as a fluoride gel and fluoride solution. The mean fluoride concentration in oral fluid was up to 10 times higher at the treated sites than at the non-treated sites. Use of a fresh toothpick or a fresh piece of dental floss in each approximal space resulted in higher values compared with using one and the same toothpick/floss for the whole dentition. An interdental brush dipped in 0.2% NaF gel and a mouthrinse with 0.2% NaF resulted in elevated fluoride concentrations at the same level as when multiple toothpicks were used. To conclude, there are large variations in the fluoride release from various brands of fluoridated toothpicks and dental flosses. Treatment with a fluoridated toothpick or a dental floss can be expected to give elevated fluoride concentrations in the approximal area up to 60 min. Another interesting method for administering fluoride in the approximal area is to use an interdental brush dipped in fluoride gel. PMID- 14763782 TI - Polymorphisms in an interferon-gamma receptor-1 gene marker and susceptibility to periodontitis. AB - Chronic marginal periodontitis is an inflammatory condition in which the supporting tissues of the teeth are destroyed. Interferon (IFN)-gamma is a cytokine that plays a pivotal role in the defense against infection, and mutations in the gene coding for the ligand binding chain (alpha, R1) of the IFN gamma receptor (IFNGR1) confer suseptibility on infections caused by poorly virulent mycobacteria. Using an intronic (CA)n polymorphic microsatellite marker within the IFNGR1 gene we investigated whether genetic polymorphisms are associated with periodontitis. In 62 periodontitis patients and 56 healthy controls we found a total of 13 polymorphisms, 11 of which were found in the periodontitis patients and 9 in the controls. Although we observed a trend towards an association with disease for allele 192, there were no significant differences in allele frequency between patients and controls. We therefore cannot find any evidence to suggest that IFNGR1, as a single dominant gene, contributes to susceptibility to periodontitis. However, in combination with the environmental risk factor, smoking, the same allelic marker was significantly associated [OR = 5.56 (1.16 75%) in an extract free from wine major volatiles. Detection limits are in the 40-300 ng/l range, well below the odor detection threshold of these compounds. Linearity (r2 > 0.996) and precision (average R.S.D. 3.5%) are satisfactory in all cases. The levels in wine of some of these lactones (gamma octa, undeca and dodecalactones) are reported by first time and results demonstrate that can be present at concentrations above or close to their corresponding odor thresholds. PMID- 14763799 TI - Solid-phase extraction versus solid-phase microextraction for the determination of chlorinated paraffins in water using gas chromatography-negative chemical ionisation mass spectrometry. AB - Solid-phase extraction (SPE) and solid-phase microextraction (SPME) were evaluated for the analysis of short-chain chlorinated paraffins (SCCPs) in water samples using gas chromatography coupled to negative chemical ionisation mass spectrometry (GC-NCI-MS). For SPE optimisation, four commercially available SPE cartridges were tested and several SPE parameters, such as the elution solvent, elution volume and breakthrough volume were studied. The best results were obtained with Varian Bond Elut-C18. In order to achieve a high selectivity in the determination of SCCPs, GC-NCI-MS was used. Quality parameters of the optimised SPE and SPME procedures were determined, and the best results were obtained for the SPE/GC-NCI-MS method with LODs of 5 and 20 ng l(-1) for tap and river water, respectively. This method was successfully applied to the analysis of SCCPs in river water samples at concentrations below the microg l(-1) level. PMID- 14763800 TI - Determination of ochratoxin A in wine using liquid-phase microextraction combined with liquid chromatography with fluorescence detection. AB - A liquid-liquid microextraction technique (LPME) has been applied to the extraction of ochratoxin A (OTA) from wine prior to its quantification by HPLC fluorescence detection. OTA was extracted from wine, through 1-octanol immobilized in the pores of a porous hollow fiber, and introduced into 1-octanol inside the fiber. Recovery was 77%. The method was adequate for quantification of OTA in wine at levels within the range 0.25-10 ng/ml with a LOD of 0.2 ng/ml, and can be a simple and inexpensive alternative to the use of inmunoaffinity columns in order to quantify OTA levels in wine. PMID- 14763801 TI - Effect of Coriolis force on counter-current chromatographic separation by centrifugal partition chromatography. AB - The effect of Coriolis force on the counter-current chromatographic separation was studied using centrifugal partition chromatography (CPC) with four different two-phase solvent systems including n-hexane-acetonitrile (ACN); tert-butyl methyl ether (MtBE)-aqueous 0.1% trifluoroacetic acid (TFA) (1:1); MtBE-ACN aqueous 0.1% TFA (2:2:3); and 12.5% (w/w) polyethylene glycol (PEG) 1000-12.5% (w/w) dibasic potassium phosphate. Each separation was performed by eluting either the upper phase in the ascending mode or the lower phase in the descending mode, each in clockwise (CW) and counterclockwise column rotation. Better partition efficiencies were attained by the CW rotation in both mobile phases in all the two-phase solvent systems examined. The mathematical analysis also revealed the Coriolis force works favorably under the CW column rotation for both mobile phases. The overall results demonstrated that the Coriolis force produces substantial effects on CPC separation in both organic-aqueous and aqueous-aqueous two-phase systems. PMID- 14763802 TI - Development of standard operation procedures for the manufacture of n-octadecyl bonded silicas as packing material in certified reference columns for reversed phase liquid chromatography. AB - The development of standard operation procedures for the manufacture of a n octadecyl bonded spherical silica packing from partially condensed tetraethoxysilane as silica source is described. The synthesis comprises five intermediate products and six synthesis steps which were examined according to their reproducibility and robustness. The results led to the optimisation of the manufacturing process for a n-octadecyl bonded silica. Correlations were drawn between the dynamic viscosity of the poly(ethoxy)siloxane (PES), the synthesis parameters, the resulting pore structural properties and particle size distribution of the silicas. Validated procedures were developed to manufacture spherical porous ultra-pure silicas with a specific surface area of 350 m2 g(-1) +/- 5% R.S.D., a specific pore volume of 1.0 ml (-1) +/- 3.7% R.S.D., an average pore diameter of 12.0 nm +/- 0.5% R.S.D. and an average particle diameter of 5 microm. Results are presented on trial batches and the final master batch which were both used as packing materials in reversed-phase liquid chromatography (RP LC) columns. The latter columns were certified and accepted as an HPLC column as reference material (BCR-722) by the European Commission, Institute for Reference Materials and Measurements (IRMM), Geel, Belgium. PMID- 14763803 TI - Characterisation of reversed-phase liquid-chromatographic columns by chromatographic tests comparing column classification based on chromatographic parameters and column performance for the separation of acetylsalicylic acid and related compounds. AB - Selection of RP-LC columns with suitable selectivity for a given analysis is difficult. For example, the European Pharmacopoeia (Ph. Eur.) and other official compendia for drug analysis only give a general description of the stationary phase in the operating procedure of a liquid chromatographic method. The need for a general test method to characterise RP-LC columns has been rising since the 1970s. A project to define a chromatographic procedure characterising RP-LC columns was started earlier. A procedure to measure test parameters was introduced and a classification of the columns, based on a minimal number of parameters, was obtained. This paper focuses on correlating the column classification with the selectivity obtained for a real separation. The separation of acetylsalicylic acid (aspirin) and related compounds was performed according to the Ph. Eur. monograph on the stationary phases previously characterised chromatographically. It was examined whether the classes of columns, determined using test parameter results, contain either suitable or unsuitable supports for the aspirin separation. The system suitability test prescribed by the Ph. Eur. in order to distinguish between suitable or unsuitable columns for this separation was also evaluated. PMID- 14763804 TI - Simultaneous analysis of amino acid and biogenic polyamines by high-performance liquid chromatography after pre-column derivatization with N-(9 fluorenylmethoxycarbonyloxy)succinimide. AB - A high-performance liquid chromatography method for the simultaneous analysis of amino acids and biogenic polyamines, using a new procedure for pre-column derivatization of amino groups with N-(9-fluorenylmethoxycarbonyloxy)succinimide is described. The separation of 20 amino acids and 4 biogenic polyamines was achieved within 32 min on a sequence of three short (50 mm) reversed-phase C18, 5 microm columns by elution buffers based on dibutylamine phosphate. The method linearity, calculated for each amino acid and polyamine, has a correlation coefficient higher than 0.991, in concentrations ranging from 0.2 to 50 microM, except for spermine and methionine, where the correlation coefficients were r = 0.984 and r = 0.979, respectively. The stability of derivatives in acidified samples at 4 degrees C and room temperature was demonstrated. The limit of quantitation was estimated to be around 50 pM in 50 microl sample injection. The repeatability of the method, expressed as R.S.D., ranged from 1.1 to 6.7%. The presented method was applied for the quantitation of amino acid and polyamine contents in beer, wine, and cell culture samples, using 2-aminoheptanoic acid or 1,7-diaminoheptane as internal standard. PMID- 14763805 TI - Enrichment of proteinaceous materials on a strong cation-exchange diol silica restricted access material: protein-protein displacement and interaction effects. AB - A study of size exclusion and enrichment of proteins employing strong cation exchange diol silica restricted access material (SCX-RAM) under saturation conditions is presented. Experiments were carried out with bacitracin, protamine, ribonuclease, lysozyme and bovine serum albumin as individual proteinaceous analytes as well as comprehensive binary mixtures and with human urine samples. Protein size dependent capacity features of the SCX-RAM column was observed. Bacitracin demonstrated the highest capacity followed by protamine while adsorption capacities of both ribonuclease and lysozyme were found smaller by a factor of 10. Applying binary protein samples occurring displacement effects were apparent: proteins with strong cationic properties displaced those already adsorbed by the bonded cation-exchange ligands. Bacitracin was displaced in all binary mixture experiments in particular by protamine. Furthermore, the binary mixtures displayed increased adsorption for some proteins due to complex formation. Lysozyme and ribonuclease showed double capacity values when paired with bacitracin. Both phenomena, displacement and enhanced adsorption occurred in the saturated state and led to changes in the urine composition during sample preparation. Injecting urine samples the relative proportions of fractions changed from 4 up to more than 20 times, due to the differences of the protein adsorption capacities on the SCX-RAM column. Analysing urine samples the SCX-RAM column provided extensive long-term stability. PMID- 14763806 TI - Isoelectric point separation of proteins by capillary pH-gradient ion-exchange chromatography. AB - In the present work, isoelectric point (pl) separation of proteins by pH-gradient ion-exchange chromatography (IEC) on packed capillary columns is demonstrated. The development of a miniaturized flow-through pH probe for reliable pH monitoring of the column effluent, which was an important technical challenge for adapting this technique to capillary dimensions, was solved by designing a low microliter per minute flow rate housing to a commercially available micro pH probe. Highly linear outlet pH-gradients within the pH range 8.5-4.0 were obtained when applying simple inexpensive buffers consisting solely of piperazine, N-methylpiperazine and imidazole on 10 cm x 0.32 mm i.d. fused silica capillaries packed with anion-exchange poly(styrene divinylbenzene)-based macroporous materials, i.e. 10 microm Mono P from Amersham Biosciences and 10 microm PL-SAX from PolymerLabs. Furthermore, when using a pH-gradient from 6.8 to 4.3, both columns were able to baseline separate the A and B genetic variants of beta-lactoglobulin, which differ with two amino acid residues only, but the PL SAX column provided almost a two-fold decrease in peak widths compared to the Mono P column. The influence of varying the buffer concentration, injection volume and column temperature on the peak widths and resolution of the beta lactoglobulins was investigated, e.g. a 100 microl sample of dilute beta lactoglobulins was injected directly on the column with practically no increase in peak width as compared to what obtained with conventional injection volumes. Finally, a pH-gradient from 6.8 to 4.3 was used to separate proteins in skimmed bovine milk on the PL-SAX column. The milk was simply diluted 1:10 (v/v) with water and filtrated before injection. PMID- 14763807 TI - Using new structurally related additive schemes in the precalculation of gas chromatographic retention indices of polychlorinated hydroxybiphenyls on HP-5 stationary phase. AB - A new additive scheme is proposed for the precalculation of gas chromatographic retention indices of complex organic compounds. The principal feature of this approach is the absence of previously calculated I increments for any structural fragments or functional groups in the molecule. Instead, arithmetical operations involving I values of simpler structural analogues of target compounds are used directly. I precalculation for polychlorinated hydroxybiphenyls (839 congeners) on the HP-5 stationary phase was chosen as one of the most important applications of the method under discussion. Such a large number of congeners cannot be obtained as reference samples and their gas chromatographic (GC)-mass spectrometric (MS) identification should therefore be based currently on precalculated I values. PMID- 14763808 TI - Capillary gas chromatography-mass spectrometry of all 93 acyclic octenes and their identification in fluid catalytic cracked gasoline. AB - The Kovats retention indices of all 93 acyclic octenes on polydimethylsiloxane and squalane as stationary phases as well as their mass spectra were measured. The means of gas chromatography-mass spectrometry (GC-MS) were used for confirmation of GC identification as well as for mass spectrometric deconvolution of the majority of gas chromatographic unseparated isomeric octene peaks. The distinction between corresponding E and Z acyclic octenes, that is either difficult or even impossible by means of GC-MS, was obtained on the basis of larger temperature coefficients of retention indices for Z isomeric octenes than for corresponding E isomers. The retention data expressed as homomorphy factors were correlated with the degree of branching, position of double bond, and position of alkyl group with respect to the double bond of acyclic octenes, and the structure-retention relationships were formulated. The 81 acyclic octenes were identified in FCC gasoline. PMID- 14763809 TI - Gas chromatographic-mass spectrometric determination of polycyclic aromatic hydrocarbons formed during the pyrolysis of phenylalanine. AB - The formation of polycyclic aromatic hydrocarbons (PAHs) during pyrolysis process of phenylalanine had been studied. Ten PAHs, including fluorene, phenanthrene, anthracene, fluoranthene, pyrene, benzo[a]anthracene, chrysene, benzo[k]fluoranthene, benzo[e]pyrene, and benzo[a]pyrene were analyzed by gas chromatography-mass spectrometry using selective ion monitoring mode. This technique offers the capability to analyze trace amounts of PAHs in phenylalanine pyrolyzates. The pyrolysis was carried out in a micro-furnace with quartz furnace liner. The injection was conducted with glass pelletizer syringe to avoid metal contamination. Qualitative results were obtained at 900 degrees C and quantitative analysis of 10 PAHs was done for 700 and 900 degrees C. PMID- 14763810 TI - Optimisation of stir bar sorptive extraction for the analysis of volatile phenols in wines. AB - An easy, fast and reliable analytical method is proposed for the determination of the concentration of volatile phenols (ethyl- and vinylphenols) in wines. The novel stir bar sorptive extraction (SBSE) technique is employed, following a simple and fast procedure that allows 15 samples to be extracted simultaneously using very small sample volume. Extracts are desorbed in a thermodesorption system (TDS) coupled on-line to a gas chromatograph-mass spectrometry system. The SBSE offers better recovery and linear regression coefficient (r2) for the four volatile phenols than solid-phase extraction (SPE). The mass spectrometric detection in selected ion monitoring mode contributes to the lower detection limit and good sensibility obtained with this method. PMID- 14763811 TI - Characterization of bile acids and fatty acids from ox bile in oil paintings by gas chromatography-mass spectrometry. AB - Characterization of ox bile, traditionally used in painting, is of interest in the fields of archaeometry and conservation and restoration of works of art. Bile acids, fatty acids (F), and cholesterol found in ox bile have been identified using a derivatization method that combines the formation of ethyl esters from the carboxylic groups and the trimethylsilyl ethers from hydroxyl groups. This method of analysis is consistent with these others proposed by the authors to analyze drying oils, proteins, and diterpenic resins usually used as binders and varnishes by the painters. Bile acids from binary samples such as animal glue/ox bile, casein/ox bile and Arabic gum/ox bile have been successfully analyzed using the proposed method. Finally, a method of analysis of mixtures of drying oil and ox bile has been also proposed attempting to quantitatively characterize samples in which ox bile was added to the drying oil for increasing the surfactant properties. PMID- 14763812 TI - Suppression of pigment interference in the gas chromatographic analysis of proteinaceous binding media in paintings with EDTA. AB - A method to suppress the interference of pigments in the analysis of proteinaceous media used in paintings is presented in this paper. This method is based on the formation of metallic ion-ethylenediaminetetraacetic acid (EDTA) complexes previous to the derivatisation process, using ethyl chloroformate (ECF), to transform the amino acids in N(O,S)-ethoxycarbonyl (EOC) ethyl esters. Test specimens, containing different proteinaceous media such as albumin, porcine gelatine and casein mixed with lead white, chalk, verdigris and raw Sienna have been prepared for carrying out this study. Different pH conditions have been probed for the different pigments studied. Values of peak area ratio of amino acids relative to the alanine, obtained using the proposed method on a series of protein-pigment test specimens, have been compared to those from specimens of pure protein in which direct method of derivatisation was applied. Finally, the method has been successfully applied to the analysis of 18th century wall paintings in which animal glue was used as binding medium. PMID- 14763813 TI - Effects of the length and modification of the separation channel on microchip electrophoresis-mass spectrometry for analysis of bioactive compounds. AB - Analyses of amino acids and peptides were performed using a quartz microchip and an interface for microchip electrophoresis-electrospray ionization mass spectrometry (MCE-ESI-MS). In MCE-ESI-MS, negative pressure caused by ESI increased band broadening and deteriorated separation. We tried to suppress the negative pressure and improve separation using a microchip with a long separation channel. Separations of peptide standards were compared using two microchips with long separation channel (58.9 mm) and short one (22.9 mm). Theoretical plate numbers and resolution were improved significantly using the former. The theoretical plate numbers of [Val4]angiotensin was 8600 using the former and 1700 using the latter. When background electrolytes of low pH were used in an uncoated quartz microchip, electrokinetic injection was difficult because of weak electroosmotic flow. The use of successive multiple ionic polymer layers coating of the microchip channel stabilized electrokinetic injection and permitted analysis of amino acids and peptides even under low pH conditions. Separation of amino acids was successfully performed using formic acid solution (pH 2.5) as background electrolyte. PMID- 14763814 TI - Purification of phospholipase D by two-phase affinity extraction. AB - An aqueous two-phase system of polyethylene glycol (PEG)-salt was used for purification of phospholipase D (PLD) from peanuts and carrots. Alginate, a known macroaffinity ligand for PLD, was incorporated in the PEG phase and resulted in 91 and 93% of the enzyme activity (from peanuts and carrots, respectively) getting partitioned in the PEG phase. The elution of the enzyme from alginate was facilitated by exploiting the fact that the latter can be reversibly precipitated in the presence of Ca2+. The enzyme was eluted from the polymer by using 0.5 M NaCl. Peanuts and carrots PLD could be purified 78- and 17-fold with 82 and 85% activity recovery, respectively. The purified enzyme from both sources gave a single band on sodium dodecyl sulfate-polyacrylamide gel (SDS-PAGE) electrophoresis. PMID- 14763815 TI - Determination of homologues of quaternary ammonium surfactants by capillary electrophoresis using indirect UV detection. AB - This investigation describes the simultaneous separation of two major non chromophoric quaternary ammonium surfactants, alkyltrimethyl- and dialkyldimethylammonium compounds (ATMACs and DADMACs, respectively), by capillary electrophoresis (CE) using indirect UV detection. The most effective separation conditions was 10 mM phosphate buffer with 57.5% tetrahydrofuran and 3 mM sodium dodecyl sulfate (SDS) at pH 4.3, and the sample hydrodynamic injection of up to 20 s at 1 psi (approximately 60 nl), and an applied voltage of 25 kV (1 psi = 6.9 kPa). Specially, the selection of an appropriate chromophore and an internal standard (I.S.) to improve the peak identification and quantitation was systematically investigated. Decylbenzyldimethyl ammonium chloride (C10-BDMA+C-) as a chromophore with 3 mM sodium dodecyl sulfate provided the best detectability for all homologues. The reproducibility of the migration time and quantitative analysis can be improved by using tetraoctyl ammonium ion as an internal standard, giving the relative standard deviation (R.S.D.) less than 0.8% for the relative migration times, and 2.5-5.5% for the relative peak areas. A good linearity of CE analysis was obtained in the range of 1.0-20 microg/ml with r2 values of above 0.999. The analysis of cationic surfactants in commercial products of hair conditioners and fabric softeners was also performed. Electrospray mass spectrometric method was applied to evaluate the CE method, and the compatible results were obtained. PMID- 14763816 TI - Simultaneous determination of vitamin E homologs in chicken meat by liquid chromatography with fluorescence detection. AB - A simple and reliable method for the simultaneous determination of all eight homologs of Vitamin E in chicken meat is described. All analytes, including the internal standard (alpha-tocopherol acetate), were eluted within 35 min and detected using their native fluorescence (295 nm excitation and 330 nm emission). Chromatography using hexane based eluent on a normal phase silica column included an initial column conditioning step to prevent irreversible adsorption of tocopherols and tocotrienols on silica. Lowest detectable levels of alpha tocopherol, gamma-tocopherol, alpha-tocotrienol, beta-tocotrienol, gamma tocotrienol and delta-tocotrienol were 0.73, 0.86, 1.0, 1.2, 1.7 and 1.3 ng, respectively. PMID- 14763817 TI - From time temperature integrator kinetics to time temperature integrator tolerance levels: heat-treated milk. AB - Six milk compounds were studied as potential intrinsic time temperature integrators (TTIs) for the assessment of heat-treated milk. These include the enzymes alkaline phosphatase and lactoperoxidase, the whey protein beta lactoglobulin and the chemical compounds hydroxymethylfurfural, lactulose and furosine. In previous research the inactivation/denaturation/formation kinetics of these compounds were analyzed under isothermal and nonisothermal conditions and evaluated for variability of the milk composition. The present paper focuses on the implementation of the TTIs. TTIs are validated with respect to microbiological indices and quality attributes, and a quantitative relationship between the denaturation, inactivation or formation of the TTIs and technological processes is established by construction of general time temperature tolerance (TTT) diagrams. In these diagrams temperature time combinations are presented, which lead to the same formation, inactivation or denaturation of TTIs, or result in the same level of microbiological destruction or quality degradation of the product. TTT-diagrams are very informative since they allow visualization of the impact of a thermal process on milk and evaluation of criteria for evaluating milk authenticity (conformity of the product with the terminology applied). Moreover, the optimum combination of temperature and time of heating may be readily deduced from these diagrams. PMID- 14763818 TI - Cellulose and glass fiber affinity membranes for the chromatographic separation of biomolecules. AB - Macroporous cellulose and glass membranes were prepared from filter paper and glass fiber filter, respectively. To enhance their stability, the cellulose membranes were crosslinked with epichlorohydrin, and the glass membranes were crosslinked with glutaraldehyde or organic bifunctional silanes. Several pathways for the modification, activation, and ligand immobilization were used and compared. For cellulose membranes, the diazotization method provided the best results, whereas the glutaraldehyde method provided the best performance for glass membranes, regarding both their stability and ligand immobilization capacity. The characterization of the membranes was made by using a triazine dye, bovine serum albumin, and trypsin as test ligands. The membrane morphologies and the uniformities of ligand distribution across the membrane cartridges were investigated. Numerous affinity ligands were immobilized onto the membranes, and the prepared affinity membranes have been used to separate or purify concanavalin A, peroxidase, protease inhibitors, globulin, fibronectin, and other biomolecules. PMID- 14763819 TI - L-glutamate enhances the expression of Thermus maltogenic amylase in Escherichia coli. AB - Escherichia coli BL21 (DE3) transformed with a thermostable Thermus maltogenic amylase (ThMA), isolated from a Gram-negative bacterium Thermus strain IM6501, grew well and efficiently produced ThMA in a complex medium but not in a chemically defined medium (DM). By supplementing L-glutamate to DM medium, both the specific growth rate and ThMA expression significantly increased. Alterations in the cellular responses of recombinant E. coli to L-glutamate were analyzed at the protein level by two-dimensional gel electrophoresis and mass spectrometry. The ppGpp synthase (RelA) was significantly reduced in cells grown with L glutamate and was consistent with the low level of ppGpp, an indicator of stringent response. On the other hand, protein chain elongation factor (EF-Tu) and manganese-containing superoxide dismutase (MnSOD), which protects cells against oxidative damage, was significantly elevated by L-glutamate supplementation. These results indicate that L-glutamate enhances ThMA expression and increases the E. coli growth rate not only by overcoming the stringent response but also by increasing the synthesis of EF-Tu and MnSOD. PMID- 14763821 TI - Combining classical, genetic, and process strategies for improved precursor directed production of 6-deoxyerythronolide B analogues. AB - A process for the production of erythromycin aglycone analogues has been developed by combining classical strain mutagenesis techniques with modern recombinant DNA methods and traditional process improvement strategies. A Streptomyces coelicolor strain expressing the heterologous 6-deoxyerythronolide B (6-dEB) synthase (DEBS) for the production of erythromycin aglycones was subjected to random mutagenesis and selection. Several strains exhibiting 2-fold higher productivities and reaching >3 g/L total macrolide aglycones were developed. These mutagenized strains were cured of the plasmid carrying the DEBS genes and a KS1 degrees mutant DEBS operon was introduced for the production of novel analogues when supplemented with a synthetic diketide precursor. The strains expressing the mutant DEBS were screened for improved 15-methyl-6-dEB production, and the best clone, strain B9, was found to be 50% more productive as compared to the parent host strain used for 15-methyl-6-dEB production. Strain B9 was evaluated in 5-L fermenters to confirm productivity in a scalable process. Although peak titers of 0.85 g/L 15-methyl-6-dEB by strain B9 confirmed improved productivity, it was hypothesized that the low solubility of 15-methyl-6-dEB limited productivity. The solubility of 15-methyl-6-dEB in water was determined to be 0.25-0.40 g/L, although higher titers are possible in fermentation medium. The incorporation of the hydrophobic resin XAD-16HP resulted in both the in situ adsorption of the product and the slow release of the diketide precursor. The resin-containing fermentation achieved 1.3 g/L 15-methyl-6-dEB, 50% higher than the resin-free process. By combining classical mutagenesis, recombinant DNA techniques, and process development, 15-methyl-6-dEB productivity was increased by over 100% in a scalable fermentation process. PMID- 14763820 TI - Thermal degradation of allicin in garlic extracts and its implication on the inhibition of the in-vitro growth of Helicobacter pylori. AB - Allicin, the main active principle related to Allium sativum chemistry, is considered to be responsible for the bacteriostatic properties of garlic. The work described here has demonstrated the direct implication of the allicin present in solvent-free garlic extracts obtained with ethanol (ethanolic garlic extract, EGE) and acetone (acetonic garlic extract, AGE) in the inhibition of the in-vitro growth of Helicobacter pylori (Hp), the bacterium responsible for serious gastric diseases such as ulcers and even gastric cancer. The evolution of allicin concentration as a function of time and temperature has been the subject of a kinetic study. The reaction order, activation energy, and preexponential factor (in accordance with Arrhenius theory) have been determined for the decomposition process of allicin in these organic media. First-order decomposition, an activation energy of 97.4 kJ/mol, and an Arrhenius preexponential factor of 8.9 x 10(10) s(-1) have been determined for allicin in EGE. For allicin in AGE the kinetic order determined was 1.5, the activation energy 184.5 kJ/mol, and the preexponential factor 3.1 x 10(24) s(-1) (mg/L)( 0.5). The presence or absence of allicin in these garlic products was found to be crucial for the inhibition of the in-vitro growth of Hp, as demonstrated by microbiological analysis for AGE. A relationship has been identified between the effectiveness and durability of the anti-Hp properties shown by AGE and the allicin content of these products. The bacteriostatic properties were active for up to 10 months if the samples were maintained at 6 degrees C. PMID- 14763822 TI - Keratinocyte growth factor-2 production in an ompT-deficient Escherichia coli K 12 mutant. AB - A variant form of Keratinocyte growth factor-2 (KGF-2) spanning amino acids A63 S208 was produced in the Escherichia coli K-12 host W3110. When the protein was purified using a standard process, the first six N-terminal amino acids were rapidly and specifically removed from the protein. This cleavage resulted in a truncated KGF-2 species (S69-S208). To circumvent this problem, guanidine-HCl was used to inhibit the putative proteolytic activity. This modified process resulted in a massive loss of protein product due to precipitation, in addition to the cost and corrosiveness of guanidine-HCl. To develop an economically feasible, scaleable, and robust process for KGF-2 production, we were tasked with identifying the protease(s) responsible for the N-terminal degradation. Experimental evidence revealed that OmpT (outer membrane protein T) was the primary protease involved in the N-terminal cleavage of the A63-S208 KGF-2 protein. Moreover, the OmpT-mediated cleavage occurred at a novel site (Arg-Ser). From this work, we show that production of the A63-S208 form of KGF-2 in an ompT deleted E. coli host nearly abolished the N-terminal cleavage issue. PMID- 14763823 TI - Cloning and functional expression of the dps gene encoding decaprenyl diphosphate synthase from Agrobacterium tumefaciens. AB - A newly isolated gene from Agrobacterium tumefaciens (A. tumefaciens), which encoded a decaprenyl diphosphate synthase, was cloned in Escherichia coli (E. coli), and its nucleotide sequence was determined. DNA sequence analysis revealed an open reading frame of 1077 bp capable of encoding a 358-amino-acid protein with a calculated isoelectric point of pH 5.16 and a molecular mass of 38 960 Da. The primary structure of the enzyme shared significant homology with prenyl diphosphate synthases from various sources. The deduced amino acid sequence included oligopeptide DDxxD aspartate-rich domains conserved in the majority of prenyl diphosphate synthases. High levels of the active enzyme were expressed in the soluble fraction and were readily purified to homogeneity by Ni-NTA chromatography. E. coli JM109 harboring the dps gene produced ubiquinone-10 in addition to endogenous ubiquinone-8, while E. coli JM109 harboring the dps gene mutated on the DDxxD domain lost the ability to produce ubiquinone-10, which suggests that the A. tumefaciens dps gene is functionally expressed in E. coli and that it encodes a decaprenyl diphosphate synthase. PMID- 14763824 TI - Model-based analysis and optimization of an ISPR approach using reactive extraction for pilot-scale L-phenylalanine production. AB - Based on experimental data from fermentation runs, as well as from L phenylalanine (l-Phe) separation studies, a simple model is presented that describes the total ISPR approach for on-line L-Phe separation. While fermentation process modeling via a macrokinetic model revealed an L-Phe inhibition constant of 20 +/- 1.35 g/L using recombinant E. coli cells, the reactive-extraction process modeling identified the L-Phe cation diffusion in the aqueous donor film and the transport of the lowly soluble carrier/L-Phe complex in the aqueous acceptor film as the most dominant transfer steps. The corresponding mass transfer coefficients were estimated as k(PheD) = 128 x 10(-7) cm/s (extraction) and k(CPheA) = 178 x 10(-5) cm/s (back-extraction). Simulation studies were performed for the total ISPR approach, which gave hints for strategies of further process optimization. PMID- 14763825 TI - Reactivity of pure Candida rugosa lipase isoenzymes (Lip1, Lip2, and Lip3) in aqueous and organic media. influence of the isoenzymatic profile on the lipase performance in organic media. AB - Three pure isoenzymes from Candida rugosa lipase (CRL: Lip1, Lip2, and Lip3) were compared in terms of their stability and reactivity in both aqueous and organic media. The combined effect of temperature and pH on their stability was studied applying a factorial design. The analysis of the response surfaces indicated that Lip1 and Lip3 have a similar stability, lower than that of Lip2. In aqueous media, Lip3 was the most active enzyme on the hydrolysis of p-nitrophenyl esters, whereas Lip1 showed the highest activity on the hydrolysis of most assayed triacylglycerides. The highest differences among isoenzymes were found in the hydrolysis of triacylglycerides. Thus, a short, medium, and long acyl chain triacylglyceride was the preferred substrate for Lip3, Lip1, and Lip2, respectively. In organic medium, Lip3 and Lip1 provided excellent results in terms of enantioselectivity in the resolution of ibuprofen (EF value over 0.90) and conversion, whereas initial esterification rate was higher for Lip3. However, the use of Lip2 resulted in lower values of conversion, enantiomeric excess, and enantioselectivity. In the case of trans-2-phenyl-1-cyclohexanol (TPCH) resolution, initial esterification rates were high except for Lip3, which also produced poor results in conversion and enantiomeric excess. The performance of the pure isoenzymes in the enantioselectivity esterification of these substrates was compared with different CRL crude preparations with known isoenzymatic content and the different results could not be explained by their isoenzymatic profile. Therefore, it can be concluded that other factors can also affect the catalysis of CRL and only the reproducibility between powders can ensure the reproducibility in synthesis reactions. PMID- 14763826 TI - Dye decolorization by manganese peroxidase in an enzymatic membrane bioreactor. AB - In the present work an enzymatic membrane reactor (EMR) for the oxidation of azo dyes by manganese peroxidase (MnP) has been developed. The configuration consisted of a stirred tank reactor coupled with an ultrafiltration membrane. The membrane allowed for most of the enzymatic activity to be recovered while both the parent dye and the degradation products could pass through. Different operational strategies (batch, fed-batch, and continuous) and parameters such as enzyme activity, H(2)O(2) feeding rate, hydraulic retention time (in continuous operation), and dye loading rate were studied. At best conditions, a continuous operation with a dye decolorization higher than 85% and minimal enzymatic deactivation was feasible for 18 days, attaining an efficiency of 42.5 mg Orange II oxidized/MnP unit consumed. PMID- 14763827 TI - Trypsin inhibition activity of heat-denatured ovomucoid: a kinetic study. AB - A kinetic study was conducted on the effect of heating in the temperature range of 75-110 degrees C on the trypsin inhibition activity of ovomucoid. Heat treatment of isolated ovomucoid resulted in a time-dependent decrease in trypsin inhibition activity that could accurately be described by a first-order kinetic model. The magnitude and the temperature dependence of the rate constants was affected by the pH during heat treatment. The heat stability of ovomucoid was the lowest at pH 7.6. Heat treatments intended to decrease the trypsin inhibition activity should therefore be carried out as soon as possible after laying, because the ovomucoid was inactivated faster at the pH of fresh egg white (pH 7.6). The presence of the other egg white constituents decreased the heat stability of ovomucoid compared to that of the model system of ovomucoid in buffer, presumably by the formation of ovomucoid-lysozyme complexes in the former. PMID- 14763828 TI - Toxic effects of chromium(VI) on anaerobic and aerobic growth of Shewanella oneidensis MR-1. AB - Cr(VI) was added to early- and mid-log-phase Shewanella oneidensis (S. oneidensis) MR-1 cultures to study the physiological state-dependent toxicity of Cr(VI). Cr(VI) reduction and culture growth were measured during and after Cr(VI) reduction. Inhibition of growth was observed when Cr(VI) was added to cultures of MR-1 growing aerobically or anaerobically with fumarate as the terminal electron acceptor. Under anaerobic conditions, there was immediate cessation of growth upon addition of Cr(VI) in early- and mid-log-phase cultures. However, once Cr(VI) was reduced below detection limits (0.002 mM), the cultures resumed growth with normal cell yield values observed. In contrast to anaerobic MR-1 cultures, addition of Cr(VI) to aerobically growing cultures resulted in a gradual decrease of the growth rate. In addition, under aerobic conditions, lower cell yields were also observed with Cr(VI)-treated cultures when compared to cultures that were not exposed to Cr(VI). Differences in response to Cr(VI) between aerobically and anaerobically growing cultures indicate that Cr(VI) toxicity in MR-1 is dependent on the physiological growth condition of the culture. Cr(VI) reduction has been previously studied in Shewanella spp., and it has been proposed that Shewanella spp. may be used in Cr(VI) bioremediation systems. Studies of Shewanella spp. provide valuable information on the microbial physiology of dissimilatory metal reducing bacteria; however, our study indicates that S. oneidensis MR-1 is highly susceptible to growth inhibition by Cr(VI) toxicity, even at low concentrations [0.015 mM Cr(VI)]. PMID- 14763829 TI - Stabilization of chloroperoxidase by polyethylene glycols in aqueous media: kinetic studies and synthetic applications. AB - Chloroperoxidase (CPO) is one of the most versatile of the heme peroxidase enzymes for synthetic applications. Despite the potential use of CPO, commercial processes have not been developed because of the low water solubility of many organic substrates of synthetic interest and the limited stability due to inactivation by H(2)O(2). CPO catalytic properties have been studied in aqueous solutions in the presence of short-chain poly(ethylene glycol)s (PEGs), and the sulfoxidation of thioanisole, as model substrate, has been investigated. The addition of PEGs allows a better substrate solubilization in the reaction mixture and the enzyme to retain more of its initial activity, with respect to pure buffer. Kinetic studies were performed to optimize the experimental conditions, and complete enantioselective conversion to the (R)-sulfoxide (ee = 99%) was observed in the presence of PEG 200 and tri(ethylene glycol). The relevant stabilization of chloroperoxidase due to the presence of PEGs allows the enzyme to convert the substrate with significant product yields even after 10 days, with a consequent increase in enzyme productivity. This is a promising result in view of industrial application of the enzyme. PMID- 14763830 TI - Rapid expression and purification of 100 nmol quantities of active protein using cell-free protein synthesis. AB - Two strategies for ATP regeneration during cell-free protein synthesis were applied to the large-scale production and single-column purification of active chloramphenicol acetyl transferase (CAT). Fed-batch reactions were performed on a 5-10 mL scale, approximately 2 orders of magnitude greater than the typical reaction volume. The pyruvate oxidase system produced 104 nmol of active CAT in a 5 mL reaction over the course of 5 h. The PANOx system produced 261 +/- 42 nmol, about 7 mg, of active CAT in a 10 mL reaction over the course of 4 h. The reaction product was purified to apparent homogeneity with approximately 70% yield by a simple affinity chromatography adsorption and elution. To our knowledge, this is the largest amount of actively expressed protein to be reported in a simple, fed-batch cell-free protein synthesis reaction. PMID- 14763831 TI - Proliferation of Lactobacillus plantarum in solid-state fermentation of oats. AB - In an attempt to introduce probiotic functionalities to breakfast cereals and similar food products, the technique of solid-state fermentation (SSF) was applied to cultivate Lactobacillus plantarum (NCIMB 8826) on oat bran and spent oats after lipid extraction by supercritical CO(2) extraction. When compared to the frequently favored submerged processes for bacterium incubation, SSF presents not only the potential of simple downstream processing but also a more natural growth environment for the target bacterium. Preliminary studies confirmed that oat bran contained balanced nutrients to support a 25-fold bacterium propagation within a range of moisture content from 50% to 58% after a 36-h cultivation. Limited hydrolysis of the raw materials by the enzyme complex from submerged incubation of Aspergillus awamori and A. oryzae to increase nutrient accessibility extended the exponential growth phase and enhanced bacterial growth by over 183-fold. The process with the most potential, however, was to simultaneously grow both fungi aerobically on the raw materials in solid state to achieve sufficient hydrolysis, followed by controlled fungal autolysis at 65 degrees C prior to anaerobic bacterium incubation. Following this process bacterium population reached a maximum of 7.3 x 10(9) cells in each gram of the fermented solids, corresponding to a 1653-fold increase from the point of inoculation. PMID- 14763832 TI - Thermodynamically controlled synthesis of amide bonds catalyzed by highly organic solvent-resistant penicillin acylase derivatives. AB - A study of various direct condensations between different amines, having very high pK values, and unmodified acyl donors has been performed. This has been possible by the use of a very stable PGA derivative. First, it has been found that the higher the cosolvent concentration, the higher the pK of the acyl donor and thus the higher the yield. Therefore, these high concentrations of cosolvents seem to be a requisite for certain enzymatic condensations. Using ethanolamine and 2-hydroxy-2-phenylethyl-amine as nucleophiles and phenyl acetic acid as the acyl donor, the increase in the diglyme concentration from 50 to 90% (v/v) permitted improvement of not only the yield (reaching values higher than 99% in both cases) but also the reaction rates (by 360- or 3-fold, respectively). However, even when using PGA preparations stabilized by multipoint covalent attachment, it was not possible to obtain these results by inactivation of the enzyme derivative. Thus, in the protection of the octylamine with phenylacetic acid in 90% diglyme, the enzymatic activity was more than 20-fold higher using the hydrophilized derivative than the glyoxyl PGA, which allowed us to obtain a yield higher than 99%. Thus, the use of hydrophilized derivatives that are very stable even in the presence of high concentrations of organic solvents opens new opportunities in the use of PGA in organic chemistry. PMID- 14763833 TI - Precursor-directed biosynthesis of novel triketide lactones. AB - Precursor-directed biosynthesis was used to produce different triketide lactones (R-TKLs) in a fermentation process. Plasmids expressing engineered versions of the first subunit of 6-deoxyerythronolide B synthase (DEBS1) fused to the terminal DEBS thioesterase (TE) were introduced into three different Streptomyces strains. The DEBS1 protein fused to TE had either an inactivated ketosynthase domain (KS1 degrees ) or a partial DEBS1 lacking module 1 but containing module 2 (M2+TE). Different synthetic precursors were examined for their effect on R-TKL production. An overproducing strain of S. coelicolor expressing the M2+TE protein was found to be best for production of R-TKLs. Racemic precursors were as effective as enantiomerically pure precursors in the fermentation process. The R group on the precursor significantly affected titer (propyl >> chloromethyl > vinyl). The R-TKLs were unstable in fermentation broth at pH 6-8. A two-phase fermentation with a pH shift was implemented to stabilize the products. The fermentation pH initially was controlled at optimal values for cell growth (pH 6.5) and then shifted to 5.5 during production. This doubled peak titers and stabilized the product. Finally, the concentration of synthetic precursor in the fermentation was optimized to improve production. A maximum titer of 500 mg/L 5 chloromethyl-TKL was obtained using 3.5 g/L precursor. PMID- 14763834 TI - Ability of different biomaterials to enantioselectively catalyze oxidation and reduction reactions. AB - We studied the ability of different biomaterials to enantioselectively catalyze oxidation or reduction reactions with the help of substrate rac-1-m or p ArCH(OH)Me and the 1-o-ArC(O)Me derivatives. Apoenzyme (NAD(P)(+)-dependent secondary alcohol dehydrogenase(NAD(P)-E)) and cofactor (NAD(P)(+)) were activated by preincubating immobilized aqueous plant leaf (e.g., young wheat leaves), cereal tissue (wheat bran), vegetable (e.g., carrot), and seaweed (e.g., wakame seaweed) solutions, and the NAD(P)-E oxidized only (R)-isomers highly enantioselectively. Thus, greater than 99% ee(s) of (S)-isomers (1m-5m and 1p-5p) can be obtained from corresponding rac-1-m or p-ArCH(OH)Me. Further, immobilized chlorella cells and immobilized baker's yeast can reduce highly stereoselectively; greater than 99% ee(s) of (S)-isomers (1o-5o) can be obtained from corresponding 1-o-ArC(O)Me. Specific use of each isomer ((S)-6 and (R)-6) with greater than 99% ee(s) of racemic-1-2-NpCH(OH)Me becomes possible through selective use of NAD(P)-E eluted from artemisia vulgaris indica leaves and young wheat leaves. We suggest that the pH of the reaction media can determine not only the direction of NAD(P)-E, toward enantioselectively catalyzed oxidation (pH > 7.0) or reduction reaction (pH < 7.0), but also the regioselective reactivity of NAD(P)-E to the substrate o- (pH < 7.0), m-, and p-substituted groups (pH > 7.0). Thus, in comparison to current biocatalysts, several biomaterials can serve as asymmetric reagent bases, providing easily obtained, low-cost natural catalysts with stereoselectivity, regioselectivity, and substrate specificity that work under mild conditions for asymmetric synthesis of organic compounds. PMID- 14763835 TI - Optimal experimental condition for hemicellulosic hydrolyzate treatment with activated charcoal for xylitol production. AB - Rice straw was hydrolyzed into a mixture of sugars using diluted H(2)SO(4). During hydrolysis, a variety of inhibitors was also produced, including acetic acid, furfural, hydroxymethylfurfural, and lignin degradation products (several aromatic and phenolic compounds). To reduce the toxic compounds concentration in the hydrolyzate and to improve the xylitol yield and volumetric productivity, rice straw hemicellulosic hydrolyzate was treated with activated charcoal under different pH values, stirring rates, contact times, and temperatures, employing a 2(4) full-factorial design. Fermentative assays were conducted with treated hydrolyzates containing 90 g/L xylose. The results indicated that temperature, pH, and stirring rate strongly influenced the hydrolyzate treatment, temperature and pH interfering with all of the responses analyzed (removal of color and lignin degradation products, xylitol yield factor, and volumetric productivity). The combination of pH 2.0, 150 rpm, 45 degrees C, and 60 min was considered an optimal condition, providing significant removal rates of color (48.9%) and lignin degradation products (25.8%), as well as a xylitol production of 66 g/L, a volumetric productivity of 0.57 g/L.h, and a yield factor of 0.72 g/g. PMID- 14763836 TI - Sequential feeding of glucose and valerate in a fed-batch culture of Ralstonia eutropha for production of poly(hydroxybutyrate-co-hydroxyvalerate) with high 3 hydroxyvalerate fraction. AB - Several important properties of poly(3-hydroxybutyric-co-3-hydroxyvaleric acids) (P(3HB-co-3HV) depend mainly on the HV unit fraction of the copolymer. Sequential and simultaneous feeding of glucose and valerate were employed to produce P(3HB co-3HV) in a fed-batch culture of Ralstonia eutropha, and the effects of feeding models on the cell growth, 3HV unit fraction, and copolymer productivity have been investigated. The sequential feeding of glucose and then valerate resulted in a cell density of 110.2 g/L, 3HV unit fraction of 62.7 mol %, and copolymer productivity of 0.56 g/(L.h), while the latter simultaneous feeding strategy never achieved the 3HV fraction of P(3HB-co-3HV) higher than 50%. A nuclear magnetic resonance study confirmed that the production of random copolymer P(3HB co-3HV) with high 3HV unit fraction was possible even with sequential feeding of glucose and valerate. PMID- 14763837 TI - Continuous production of mixed lactic starters containing probiotics using immobilized cell technology. AB - The production of a mixed lactic culture containing Lactococcus lactis subsp. lactis biovar. diacetylactis MD and Bifidobacterium longum ATCC 15707 was studied during a 17-day continuous immobilized-cell culture at different temperatures between 32 and 37 degrees C. The two-stage fermentation system was composed of a first reactor (R1) containing cells of the two strains separately immobilized in kappa-carrageenan/locust bean gum gel beads and a second reactor (R2) operated with free cells released from the first reactor. The system allowed continuous production of a concentrated mixed culture with a strain ratio whose composition depended on temperature and fermentation time. A stable mixed culture (with a 22:1 ratio of L. diacetylactis and B. longum) was produced at 35 degrees C in the effluent of R2, whereas the mixed culture was rapidly unbalanced in favor of B. longum at a higher temperature (37 degrees C) or L. diacetylactis at a lower temperature (32 degrees C). Strain redistribution in beads originally immobilizing pure cultures of L. diacetylactis or B. longum was observed. At the end of culture, the strain ratio (7:1 L. diacetylactis/B. longum) in bulk bead samples was similar to that of individual beads. The determination of the spatial distribution of the two strains in gel beads by immunofluorescence and confocal laser-scanning microscopy showed that bead cross-contamination was limited to a 100 microm peripheral layer. Data from this study validate a previous model for population dynamics and cell release in gel beads during mixed immobilized-cell cultures. PMID- 14763838 TI - Application of a MTT assay for screening nutritional factors in growth media of primary sponge cell culture. AB - Marine sponges (Porifera) are producers of the largest variety of bioactive compounds among benthic marine organisms. In vitro culture of marine sponge cells has been proposed for the sustainable production of these pharmacologically interesting compounds from marine sponges but with limited success. The development of a suitable growth medium is an essential prerequisite for sponge cells grown in vitro. The MTT (3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide) assay was adapted to screen for potential nutritional factors in formulating a growth medium for primary cell culture of Suberites domuncula. In 96-well plates, the effects of nutritional factors including glutamine, pyruvate, iron citrate, silicon, RPMI 1640, and Marine Broth 2216 on the viable cell density were examined in primary cell culture of S. domuncula 36 h after inoculation. Ferric iron (Fe(3+)) and pyruvate were found to significantly improve cell viability in a dose-dependent manner. Silicon and glutamine showed limited improvements at certain concentrations. The supplement of RPMI 1640 and Marine Broth 2216 did not increase cell viability. As a result, several improved media able to maintain higher cell viability in a short-term culture of primary sponge cells could be formulated. PMID- 14763839 TI - Invertase-lipid biocomposite films: preparation, characterization, and enzymatic activity. AB - The formation of biocomposite films of the industrially important enzyme invertase and fatty lipids under enzyme-friendly conditions is described. The approach involves a simple beaker-based diffusion protocol wherein invertase diffuses into the cationic lipid octadecylamine during immersion of the lipid film in the enzyme solution. Entrapment of invertase in the octadecylamine film is highly pH-dependent, underlining the role of attractive electrostatic interactions between the enzyme and the lipid in the biocomposite film formation. The kinetics of formation of the enzyme-lipid biocomposites has been studied by quartz crystal microgravimetry (QCM) measurements. The stability of the enzyme in the lipid matrix was confirmed by fluorescence spectroscopy and biocatalytic activity measurements. The biocatalytic activity of the invertase-lipid biocomposite films was comparable to that of the free enzyme in solution and showed marginally higher temperature stability. Particularly exciting was the excellent reuse characteristics of the biocomposite films, indicating potential industrial application of these films. PMID- 14763840 TI - A hybrid model framework for the optimization of preparative chromatographic processes. AB - An optimization framework based on the use of hybrid models is presented for preparative chromatographic processes. The first step in the hybrid model strategy involves the experimental determination of the parameters of the physical model, which consists of the full general rate model coupled with the kinetic form of the steric mass action isotherm. These parameters are then used to carry out a set of simulations with the physical model to obtain data on the functional relationship between various objective functions and decision variables. The resulting data is then used to estimate the parameters for neural network-based empirical models. These empirical models are developed in order to enable the exploration of a wide variety of different design scenarios without any additional computational requirements. The resulting empirical models are then used with a sequential quadratic programming optimization algorithm to maximize the objective function, production rate times yield (in the presence of solubility and purity constraints), for binary and tertiary model protein systems. The use of hybrid empirical models to represent complex preparative chromatographic systems significantly reduces the computational time required for simulation and optimization. In addition, it allows both multivariable optimization and rapid exploration of different scenarios for optimal design. PMID- 14763841 TI - Separation of lactic acid from acetic acid using a four-zone SMB. AB - A simulated moving bed (SMB) process has been developed to separate l-(+)-lactic acid from acetic acid, a major impurity in the fermentation broth of Lactobacillus rhamnosus. Poly(4-vinylpyridine) resin (PVP) was selected as the adsorbent. Adsorption isotherms and mass transfer parameters of the organic acids were estimated from single-column frontal tests. Experimental results show that the Langmuir isotherms obtained from the frontal tests can be used in the design of an SMB process to achieve 99.9% purity and over 93% yield of lactic acid. The column profiles and effluent histories, however, deviate from rate model predictions based on the Langmuir isotherms. They agree more closely with the predictions based on a modified Langmuir isotherm for lactic acid. The standing wave design method for systems with modified Langmuir isotherms is developed in this study. Rate model simulations show that the process based on the modified design method can achieve high purity (>99.9%) and high yield (>99.9%). For this nonlinear system, accurate isotherm model and model parameters are needed in the design, and the zone flow rates must be closely monitored and controlled in order to ensure high purity and high yield in the SMB process. PMID- 14763842 TI - Modeling two-component isoelectric focusing buffers in a vortex-stabilized electrophoresis apparatus. AB - Defined buffer systems have been considered by researchers as replacements for synthetic carrier ampholyte mixtures in IEF in order to reduce both costs and deleterious protein-ampholyte interactions. In this paper, a mathematical model for the behavior of two-component pH gradients during IEF is applied to a vortex stabilized electrophoresis apparatus. Equations for mass conservation, molar fluxes, and the electric field describe the behavior of the components in the electric field. Equilibrium constants are used in the model to account for interconversion between the positive, negative, and neutral states for each component. The model was applied to predict pH gradients using two different defined buffer systems. The model's predictions fell within the 95% confidence interval of a least-squares fit to the experimental pH gradients except in the regions near the ends of the pH gradients, where large pH excursions occurred during the experiments. PMID- 14763843 TI - Effect of inhibitors released during steam-explosion treatment of poplar wood on subsequent enzymatic hydrolysis and SSF. AB - Steam-exploded (SE) poplar wood biomass was hydrolyzed by means of a blend of Celluclast and Novozym cellulase complexes in the presence of the inhibiting compounds produced during the preceding steam-explosion pretreatment process. The SE temperature and time conditions were 214 degrees C and 6 min, resulting in a log R(0) of 4.13. In enzymatic hydrolysis tests at 45 degrees C, the biomass loading in the bioreactor was 100 g(DW)/L (dry weight) and the enzyme-to-biomass ratio 0.06 g/g(DW). The enzyme activities for endo-glucanase, exo-glucanase, and beta-glucosidase were 5.76, 0.55, and 5.98 U/mg, respectively. The inhibiting effects of components released during SE (formic, acetic, and levulinic acids, furfural, 5-hydroxymethyl furfural (5-HMF), syringaldehyde, 4-hydroxy benzaldehyde, and vanillin) were studied at different concentrations in hydrolysis runs performed with rinsed SE biomass as model substrate. Acetic acid (2 g/L), furfural, 5-HMF, syringaldehyde, 4-hydroxybenzaldehyde, and vanillin (0.5 g/L) did not significantly effect the enzyme activity, whereas formic acid (11.5 g/L) inactivated the enzymes and levulinic acid (29.0 g/L) partially affected the cellulase. Synergism and cumulative concentration effects of these compounds were not detected. SSF experiments show that untreated SE biomass during the enzymatic attack gives rise to a nonfermentable hydrolysate, which becomes fermentable when rinsed SE biomass is used. The presence of acetic acid, vanillin, and 5-HMF (0.5 g/L) in SSF of 100 g(DW) /L biomass gave rise to ethanol yields of 84.0%, 73.5%, and 91.0% respectively, with respective lag phases of 42, 39, and 58 h. PMID- 14763844 TI - A model for the salt effect on adsorption equilibrium of basic protein to dye ligand affinity adsorbent. AB - A model describing the salt effect on adsorption equilibrium of a basic protein, lysozyme, to Cibacron Blue 3GA-modified Sepharose CL-6B (CB-Sepharose) has been developed. In this model, it is assumed that the presence of salt causes a fraction of dye-ligand molecules to lodge to the surface of the agarose gel, resulting from the induced strong hydrophobic interaction between dye ligand and agarose matrix. The salt effect on the lodging of dye-ligand is expressed by the equilibrium between salt and dye-ligand. For the interactions between protein and vacant binding sites, stoichiometric equations based either on cation exchanges or on hydrophobic interactions are proposed since the CB dye can be regarded as a cation exchanger contributed by the sulfonate groups on it. Combining with the basic concept of steric mass-action theory for ion exchange, which considers both the multipoint nature and the macromolecular steric shielding of protein adsorption, an explicit isotherm for protein adsorption equilibrium on the dye ligand adsorbent is formulated, involving salt concentration as a variable. Analysis of the model parameters has yielded better understanding of the mechanism of salt effects on adsorption of the basic protein. Moreover, the model predictions are in good agreement with the experimental data over a wide range of salt and ligand concentrations, indicating the predictive nature of the model. PMID- 14763845 TI - Principal component analysis of nonlinear chromatography. AB - Principal component analysis (PCA) has been used for the modeling of nonlinear chromatography under overload conditions. A 10-fold range of crude erythromycin samples were loaded onto columns with different stationary-phase chemistries (2 polystyrene, 1 methacrylate) in direct proportion to the bed volumes. The elution profiles indicated slightly concave isotherms for the polystyrene supports and a convex Langmuirian isotherm for the methacrylic support used. The principal component models accounted for over 98% of the original variance in the data for all three columns and were able to give excellent models of complete chromatograms in the absence of first-principle models or physicochemical data. Correlations between sample mass and the principal component scores were made for each that were consistent for the column types despite the different geometries and stationary phases. Linear relationships with high correlation coefficients were observed when the scores of the same principal component were compared between columns. Such correlations offer considerable potential for modeling of nonlinear chromatography. PMID- 14763846 TI - A new metal chelate affinity adsorbent for cytochrome C. AB - We have prepared a novel metal-chelate adsorbent utilizing N-methacryloyl-L histidine methyl ester (MAH) as a metal-chelating ligand. MAH was synthesized by using methacryloyl chloride and l-histidine methyl ester dihydrochloride. Spherical beads with an average diameter of 75-125 microm were produced by suspension polymerization of 2-hydroxyethyl methacrylate (HEMA) and MAH carried out in an aqueous dispersion medium. Then, Cu(2+) ions were chelated directly on the chelating beads. Cu(2+)-chelated beads were used in the adsorption of cytochrome c (cyt c) from aqueous solutions. The maximum cyt c adsorption capacity of the Cu(2+)-chelated beads (658.2 micromol/g Cu(2+) loading) was found to be 31.7 mg/g at pH 10 in phosphate buffer. The nonspecific cyt c adsorption on the naked PHEMA beads was 0.2 mg/g. Cyt c adsorption increased with increasing Cu(2+) loading. Cyt c adsorption capacity was demonstrated for the buffer types with the effects in the order phosphate > HEPES > MOPS > MES > Tris-HCl. Cyt c molecules could be adsorbed and desorbed five times with these adsorbents without noticeable loss in their cyt c adsorption capacity. PMID- 14763847 TI - Collapse temperature of freeze-dried Lactobacillus bulgaricus suspensions and protective media. AB - Optimization of the freeze-drying process needs to characterize the physical state of frozen and dried products. A protocol to measure the collapse temperature of complex biological media such as concentrated lactic acid bacteria using freeze-drying microscopy was first elaborated. Afterward, aqueous solutions of one or several components as well as concentrated lactic acid bacterial suspensions were analyzed in order to study how the structure of these materials is degraded during freeze-drying. A similar behavior toward collapse was observed for all aqueous solutions, which was characterized by two temperatures: the "microcollapse" temperature (T(microc), beginning of a local loss of structure) and the "collapse" temperature (T(c), beginning of an overall loss of structure). For aqueous solutions, these two temperatures were close, differing by less than 3 degrees C. Nevertheless, when lactic acid bacteria were added to aqueous solutions, the collapse temperatures increased. Moreover, the interval between microcollapse and collapse temperatures became larger. Lactic acid bacterial cells gave a kind of "robustness" to the freeze-dried product. Finally, comparing glass transition, measured by differential scanning calorimetry (DSC) and collapse temperature for aqueous solutions with noncrystallizable solutes, showed that these values belonged to the same temperature range (differing by less than 5 degrees C). As suggested in the literature, the glass transition temperature can thus be used as a first approximation of the collapse temperature of these media. However, for lactic acid bacterial suspensions, because the difference between collapse and glass transition temperatures was about 10 degrees C, this approximation was not justified. An elegant physical appearance of the dried cakes and an acceptable acidification activity recovery were obtained, when applying operating conditions during freeze-drying in vials that allowed the product temperature to be maintained during primary drying at a level lower than the collapse temperature of lactic acid bacterial suspensions. Consequently, the collapse temperature T(c) was proposed as the maximal product temperature preserving the structure from macroscopic collapse and an acceptable biological activity of cells. PMID- 14763848 TI - Evaluation of oxygen mass transfer in Aspergillus niger fermentation using data reconciliation. AB - Fermentation experiments using Aspergillus niger result in a very viscous broth due to the growth of filamentous microorganism. For viscous fermentation processes, it is difficult to estimate with confidence the volumetric oxygen mass transfer coefficient (K(L)a), which can be used for scale-up or design of bioreactors. In the present study, four methods based on dynamic and stationary approaches were used to measure K(L)a throughout the fermentation. Data reconciliation was used to obtain a more reliable and consistent K(L)a. The K(L)a value obtained by a data reconciliation technique was found to be more reliable since it takes into consideration both the reliability of all measured variables and the accuracy of all mass balance equations. PMID- 14763850 TI - Modification of high lignin content kraft pulps with laccase to improve paper strength properties. 1. Laccase treatment in the presence of gallic acid. AB - Laccase was reacted with gallic acid in the presence of a high-kappa (91) kraft pulp. The result was a modified pulp with 34%, 20%, and 72% improvements in burst, tensile, and wet tensile strength compared to untreated control samples. Fully bleached pulps were not responsive to the laccase treatment, indicating lignin was the major target for the fiber modification. The results indicate that the strength increases were a combined effect of improvements of hydrogen bonding between fibers and creation of phenoxy radical cross-links within the sheet. PMID- 14763849 TI - Effects of various sugars added to growth and drying media upon thermotolerance and survival throughout storage of freeze-dried Lactobacillus delbrueckii ssp. bulgaricus. AB - The aim of this research effort was to investigate the role of various sugar substrates in the growth medium upon thermotolerance and upon survival during storage after freeze-drying of Lactobacillus bulgaricus. Addition of the sugars tested to the growth medium, and of these and sorbitol to the drying medium (skim milk) was investigated so as to determine whether a relationship exists between growth and drying media, in terms of protection of freeze-dried cells throughout storage. The lowest decrease in viability of L. bulgaricus cells after freeze drying was obtained when that organism was grown in the presence of mannose. However, L. bulgaricus clearly survived better during storage when cells had been grown in the presence of fructose, lactose or mannose rather than glucose (the standard sugar in the growth medium). A similar effect could not be observed in terms of thermotolerance; in this case, the growth medium supplemented with lactose was found to yield cells bearing the highest heat resistance. Supplementation of the drying medium with glucose, fructose, lactose, mannose or sorbitol led in most cases to enhancement of protection during storage, to a degree that was growth medium-dependent. PMID- 14763851 TI - Affinity purification of lipid vesicles. AB - We present a novel column chromatography technique for recovery and purification of lipid vesicles, which can be extended to other macromolecular assemblies. This technique is based on reversible binding of biotinylated lipids to monomeric avidin. Unlike the very strong binding of biotin and biotin-functionalized molecules to streptavidin, the interaction between biotin-functionalized molecules and monomeric avidin can be disrupted effectively by ligand competition from free biotin. In this work, biotin-functionalized lipids (biotin-PEG-PE) were incorporated into synthetic lipid vesicles (DOPC), resulting in unilamellar biotinylated lipid vesicles. The vesicles were bound to immobilized monomeric avidin, washed extensively with buffer, and eluted with a buffer supplemented with free biotin. Increasing the biotinyl lipid molar ratio beyond 0.53% of all lipids did not increase the efficiency of vesicle recovery. A simple adsorption model suggests 1.1 x 10(13) active binding sites/mL of resin with an equilibrium binding constant of K = 1.0 x 10(8) M(-1). We also show that this method is very robust and reproducible and can accommodate vesicles of varying sizes with diverse contents. This method can be scaled up to larger columns and/or high throughput analysis, such as a 96-well plate format. PMID- 14763852 TI - Application of vortex flow adsorption technology to intein-mediated recovery of recombinant human alpha1-antitrypsin. AB - Vortex flow is a secondary flow pattern that appears above a critical rotation rate in the annular gap between an inner rotating solid cylinder and an outer stationary cylindrical shell. By suspending adsorbent resin in the vortices, a novel unit operation, vortex flow adsorption (VFA), is created. In VFA, the rotation of the inner cylinder facilitates the fluidization of the adsorbent resin. Similar to expanded bed processes, VFA has high fluid voidage so that it can be used to recover biochemical products directly from fermentation broths or cell homogenates without removing cells or cell debris first. In this study, recombinant human alpha1-antitrypsin (alpha1-AT) was expressed in Escherichia coli as a fusion with a modified intein containing a chitin-binding domain. Therefore, the fusion protein can be recovered by chitin resin affinity adsorption. The intein can be induced to undergo in vitro peptide bond cleavage to specifically release alpha1-AT from the bound fusion protein. The capture efficiency of the fusion protein, 26.2%, was obtained in the VFA process. In addition, the specific activity of alpha1-AT was dramatically improved from 0.3 to 205.2 EIC/(mg total protein) after adsorption and cleavage. Therefore, vortex flow adsorption is an integrative technology to combine the primary clarification, concentration, and purification steps in conventional downstream processing into a single unit operation to efficiently recover and purify biochemical products. PMID- 14763853 TI - Solid-phase refolding of cyclodextrin glycosyltransferase adsorbed on cation exchange resin. AB - Expression with a fusion partner is now a popular scheme to produce a protein of interest because it provides a generic tool for expression and purification. In our previous study, a strong polycationic tail has been harnessed for an efficient purification scheme. Here, the same polycation tail attached to a protein of interest is shown to hold versatility for a solid-phase refolding method that utilizes a charged adsorbent as a supporting material. Cyclodextrin glycosyltransferase (CGTase) fused with 10 lysine residues at the C-terminus (CGTK10ase) retains the ability to bind to a cation exchanger even in a urea denatured state. When the denatured and adsorbed CGTK10ase is induced to refold, the bound CGTK10ase aggregates little even at a g/L range. The renatured CGTK10ase can also be simply recovered from the solid support by adding high concentration of NaCl. The CGTK10ase refolded on a solid support retains specific enzyme activity virtually identical to that of the native CGTK10ase. Several factors that are important in improving the refolding efficiency are explored. Experimental results indicate that nonspecific electrostatic interactions between the charge of the ion exchanger and the local charge of CGTase other than the polycationic tag should be reduced to obtain higher refolding yield. The solid phase refolding method utilizing a strong polycationic tag resulted in a remarkable increase in the refolding performance. Taken together with the previous report in which a series of polycations were explored for efficient purification, expression of a target protein fused with a strong polycation provides a straightforward protein preparation scheme. PMID- 14763854 TI - New cationic exchanger support for reversible immobilization of proteins. AB - New tailor-made cationic exchange resins have been prepared by covalently binding aspartic-dextran polymers (e.g. MW 15 000-20 000) to porous supports (aminated agarose and Sepabeads). More than 80% of the proteins contained in crude extracts from Escherichia coli and Acetobacter turbidans have been strongly adsorbed on these porous materials at pH 5. This interaction was stronger than in conventional carboxymethyl cellulose (e.g., at pH 7 and 25 degrees C, all proteins previously adsorbed at pH 5 were released from carboxymethyl cellulose, whereas no protein was released from the new supports under similar conditions). Ionic exchange properties of such composites were strongly dependent on the size of the aspartic-dextran polymers as well as on the exact conditions of the covalent coating of the solids with the polymer (optimal conditions: 100 mg aspartic-dextran 20 000/(mL of support); room temperature). Finally, some industrially relevant enzymes (Kluyveromices lactis, Aspergillus oryzae, and Thermus sp. beta-galactosidases, Candida antarctica B lipase, and bovine pancreas trypsin and chymotrypsin) have been immobilized on these supports with very high activity recovery and immobilization rates. After enzyme inactivation, the enzyme can be fully desorbed from the support and the support could be reused for several cycles. PMID- 14763855 TI - Distribution of cells between solid/liquid and liquid/liquid interfaces. AB - The use of aqueous two-phase systems (ATPSs) and each system's individual phase forming species to prevent Streptococcus sanguis attachment onto hydroxyapatite discs was explored. The strategy that we followed was to attach the cells to a solid surface in the presence of an additional interface. Conditions under which, simultaneously, the phase-forming species form two phases and the cells proliferate were identified. Growth curves were constructed in the presence of various polymers and salts commonly used to prepare ATPSs. Several aqueous two phase systems were selected such that bacterial growth was comparable to that observed in pure medium. Cells were allowed to attach to hydroxyapatite discs for 7 days in the presence of varying concentrations of media, media with polymer, media with salt, and media with ATPS. Streptococcus sanguis attachment to the disks was evaluated by scanning electron microscopy. The addition of a PEG/Na(2)SO(4) ATPS to high concentrations of yeast-tryptone (YT) media (>65%) and of a PEG/MgSO(4) ATPS to nutrient-limited media reduces surface coverage of S. sanguis to less than 10%. Comparison of the attachment levels for the systems containing PEG/Na(2)SO(4) to media containing the individual phase-forming species and to the YT reference systems indicated that nutrient availability did not affect attachment. PMID- 14763856 TI - Simulating the embolization of blood vessels using magnetic microparticles and acupuncture needle in a magnetic field. AB - Computer models were developed to simulate the capture and subsequent deposition of magnetic microparticles (MMPs) in a blood vessel adjacent to a ferromagnetic wire (e.g., acupuncture needle) magnetized by a uniform external magnetic field. Process parameter conditions were obtained to enable optimal capture of MMPs into the deposit. It was found that the maximum capture distance of the MMPs was within 0.5-2.0 mm when the particles were superparamagnetic and had large size (>1.0 microm) and relative large flow rates (2.5-5.0 cm/s) as in a healthy artery. It was also found that the deposits were asymmetrical and that their size was between 1.0 and 2.0 mm. For the case of lower flow rates as can be found in a tumor (<1.0 mm/s) and using small magnetite particles (0.25-2.0 microm) the maximum capture distance was larger, ranging between approximately 0.5 and 6.4 mm, depending on the blood flow rate, the radius of wire, and particle clustering. The range of embolization (deposition) in this later case was between 0.5 and 5.9 mm. The potential of this technique to generate MMPs deposits to embolize blood vessels inhibiting the blood supply and thus facilitating necrosis of tumors located deep within the patient (3-7 cm) is discussed. PMID- 14763857 TI - Effects of three-dimensional culturing in a fibrous matrix on cell cycle, apoptosis, and MAb production by hybridoma cells. AB - The effects of culturing hybridoma cells in a three-dimensional (3-D) poly(ethylene terephthalate) (PET) fibrous matrix on cell cycle, apoptosis, metabolism, and monoclonal antibody (MAb) production were evaluated by comparing with two-dimensional (2-D) culturing on microcarrier and multiwell plate surfaces. The percentage of cells in the G1/G0 phase increased during the long term culturing period of approximately 4 weeks. Compared to the 2-D culture systems, cells grown in 3-D matrices had higher MAb productivity for long-term culture. Decreasing serum content in the culture medium increased both MAb productivity and apoptosis. However, the 3-D culture had a greater increase in MAb productivity and a much lower apoptotic rate than the 2-D culture, especially at 0% serum. Most cells in the 3-D fibrous matrix formed large aggregates and were smaller than cells grown on a 2-D surface or in suspension. The smaller cell size allowed cells to survive better in the high-cell-density environment. The fibrous matrix also selectively retained healthy, nonapoptotic cells. These results suggested that the 3-D fibrous matrix contributed to growth arrest, protected cells to better resist low-serum environments, and reduced apoptosis, all of which contributed to the high viable cell density and volumetric MAb productivity in the long-term 3-D culture. PMID- 14763858 TI - Development of a microscale cell culture analog to probe naphthalene toxicity. AB - Prediction of human response to drugs or chemicals is difficult as a result of the complexity of living organisms. We describe an in vitro model that can realistically and inexpensively study the adsorption, distribution, metabolism, elimination, and potential toxicity (ADMET) of chemicals. A microscale cell culture analog (microCCA) is a physical replica of the physiologically based pharmacokinetics (PBPK) model. Such a microfabricated device consists of a fluidic network of channels to mimic the circulatory system and chambers containing cultured mammalian cells representing key functions of animal "organ" systems. This paper describes the application of a two-cell system, four-chamber microCCA ("lung"-"liver"-"other tissue"-"fat") device for proof-of-concept study using naphthalene as a model toxicant. Naphthalene is converted into reactive metabolites (i.e., 1,2-naphthalenediol and 1,2-naphthoquinone) in the "liver" compartment, which then circulate to the "lung" depleting glutathione (GSH) in lung cells. Such microfabricated in vitro devices are potential human surrogates for testing chemicals and pharmaceutics for toxicity and efficacy. PMID- 14763859 TI - Inhibition of apoptosis by a Bombyx mori gene. AB - An apoptosis-inhibiting component of silkworm hemolymph, isolated and characterized in our previous study, showed 95% N-terminal amino acid sequence homology with one of the 30K proteins, a group of structurally related proteins. The 30K protein was expressed in mammalian HEK293 cells and CHOK1 cells by transfection with 30Kc6. The expression of 30Kc6 inhibited apoptosis comparably to that of whole silkworm hemolymph, indicating that both intracellular expression and external supplementation inhibited apoptosis. The expression of 30Kc6 resulted in lower intracellular activity for caspase 3. However, the results of in vitro assay of caspase 3 show that the 30Kc6 protein does not inhibit caspase 3 activity. This indicates that the 30Kc6 protein inhibits the apoptosis by working in a further upstream event than caspase 3 activation. PMID- 14763860 TI - Structural and thermal stability characterization of Escherichia coli D galactose/D-glucose-binding protein. AB - The effect of temperature and glucose binding on the structure of the galactose/glucose-binding protein from Escherichia coli was investigated by circular dichroism, Fourier transform infrared spectroscopy, and steady-state and time-resolved fluorescence. The data showed that the glucose binding induces a moderate change of the secondary structure content of the protein and increases the protein thermal stability. The infrared spectroscopy data showed that some protein stretches, involved in alpha-helices and beta strand conformations, are particularly sensitive to temperature. The fluorescence studies showed that the intrinsic tryptophanyl fluorescence of the protein is well represented by a three exponential model and that in the presence of glucose the protein adopts a structure less accessible to the solvent. The new insights on the structural properties of the galactose/glucose-binding protein can contribute to a better understanding of the protein functions and represent fundamental information for the development of biotechnological applications of the protein. PMID- 14763861 TI - The design and fabrication of three-chamber microscale cell culture analog devices with integrated dissolved oxygen sensors. AB - Whole animal testing is an essential part in evaluating the toxicological and pharmacological profiles of chemicals and pharmaceuticals, but these experiments are expensive and cumbersome. A cell culture analog (CCA) system, when used in conjunction with a physiologically based pharmacokinetic (PBPK) model, provides an in vitro supplement to animal studies and the possibility of a human surrogate for predicting human response in clinical trials. A PBPK model mathematically simulates animal metabolism by modeling the absorption, distribution, metabolism, and elimination kinetics of a chemical in interconnected tissue compartments. A CCA uses mammalian cells cultured in interconnected chambers to physically represent the corresponding PBPK. These compartments are connected by recirculating tissue culture medium that acts as a blood surrogate. The purpose of this article is to describe the design and basic operation of the microscale manifestation of such a system. Microscale CCAs offer the potential for inexpensive, relatively high throughput evaluation of chemicals while minimizing demand for reagents and cells. Using microfabrication technology, a three-chamber ("lung"-"liver"-"other") microscale cell culture analog (microCCA) device was fabricated on a 1 in. (2.54 cm) square silicon chip. With a design flow rate of 1.76 microL/min, this microCCA device achieves approximate physiological liquid to-cell ratio and hydrodynamic shear stress while replicating the liquid residence time parameters in the PBPK model. A dissolved oxygen sensor based on collision quenching of a fluorescent ruthenium complex by oxygen molecules was integrated into the system, demonstrating the potential to integrate real-time sensors into such devices. PMID- 14763862 TI - Influence of temperature and pH on the nonenzymatic reduction of triphenyltetrazolium chloride. AB - The effects of temperature and pH on the nonenzymatic (chemical) reduction of triphenyltetrazolium chloride (TTC) to triphenyl formazan (TF) in cheese whey and municipal solid waste compost samples were studied. Ten different incubation temperatures and 13 pH levels were tested. The study showed that the TTC could be reduced nonenzymatically at high temperatures and/or under alkaline pH conditions. The nonenzymatic TTC reduction was observed at pH values greater than 9.5 and 11.0 for the cheese whey and compost, respectively. The TTC chemical reduction rate followed the same trend in both media. The TF content increased with increasing the pH value, reaching its maximum at a pH of 12, then decreased and was not detected at a pH of 13. The TTC was also reduced nonenzymatically at temperatures higher than 70 and 85 degrees C for cheese whey and compost, respectively. Evaporation did not seem to have any significant effect on the TTC chemical reduction since less than 3% of water content was lost at a temperature of 100 degrees C. It was noticed that the TF yield in cheese whey samples was higher than that in compost samples. This was due to the higher moisture content of cheese whey and the presence of copper in the compost samples, which reacted chemically with the TF causing reduction in the red color. For a given incubation period, the effect of pH on the TTC chemical reduction was more significant than the effect of incubation temperature (at a 2 h incubation period, 57.5% and 17.9% of the TTC were chemically reduced at a pH of 12 compared to 10.9% and 7.7% at an incubation temperature of 100 degrees C, for cheese whey and compost, respectively). Among the six metals tested (Ca, Cu, K, Na, Ni, and Zn) only Cu affected the color intensity of the TF. The activation energy of the TTC chemical reduction was 168,808 and 239,102 J/mol in cheese whey and municipal solid compost, respectively. For dehydrogenase activity measurement, the pH of the samples and the incubation temperature should not be higher than 9 and 60 degrees C in order to ensure that the TTC reduction is caused only by the biochemical reaction. Measuring the color intensity of TF in waste samples that contain copper could give misleading results as a result of the formation of formazan copper complex, which reduces the red color. PMID- 14763863 TI - Rapid titer determination of baculovirus by quantitative real-time polymerase chain reaction. AB - Titer determination is a prerequisite for the study of viruses. However, the current available methods are tedious and time-consuming. To improve the efficiency of titer determination, we have developed a rapid and simple method for the routine detection of baculovirus titers using a quantitative real-time PCR. This method is based on the amplification of approximately 150-bp fragments located in the coding regions of selected genes. The PCR was found to be quantitative in a range of 10(3) to 10(9) virus particles per 200 microL of supernatant, and the results were closely correlated with titers detected from 50% tissue culture infectious doses (TCID(50)) of baculovirus. This quantitative real-time PCR requires only 30 min to perform, and the entire titer determination can be accomplished within 1 h without the need for cell seeding or further virus dilution and infection. Because this technology is easy to operate, generates data with high precision, and most importantly is very quick, it will certainly be broadly applied for titer determination of baculoviruses in the future. PMID- 14763864 TI - Evaluation of multiwavelength culture fluorescence for monitoring the aroma compound 4-hydroxy-2(or 5)-ethyl-5(or 2)-methyl-3(2H)-furanone (HEMF) production. AB - Fluorescence spectra of a 4-hydroxy-2(or 5)-ethyl-5(or 2)-methyl-3(2H)-furanone (HEMF) fermentation culture broth were combined with measurable process variables for off-line and on-line process monitoring. Culture broth fluorescence in UV and visible ranges was acquired by a fiber optic LCD array spectrometer. Process dynamics was followed on-line using a fiber optic probe attached to an external recirculation loop of the bioreactor. Partial least squares and stepwise regression methods were used to correlate measurable process parameters with the components of the fluorescence spectra. Both methods provided adequate approximation of yeast density, HEMF, glucose, and ethanol concentrations from fluorescence spectra. HEMF production was observed during the oxido-reductive growth phase when there was a lack of measurable oxygen in the culture broth and an excess of glucose. The addition of glucose resulted in the rapid production of HEMF and other metabolite intermediates such as ethanol, acetate, and glycerol. PMID- 14763865 TI - Characterization of nerolidol biotransformation based on indirect on-line estimation of biomass concentration and physiological state in batch cultures of Aspergillus niger. AB - Biotransformation of the sesquiterpenoid trans-nerolidol by Aspergillus niger has previously been investigated as a method for the formation of 12-hydroxy-trans nerolidol, a precursor in the synthesis of the industrially interesting flavor alpha-sinensal. We characterized biotransformations of cis-nerolidol, trans nerolidol, and a commercially available cis/trans-nerolidol mixture in repeated batch cultures of A. niger grown in computer-controlled bioreactors. On-line quantification of titrant addition in pH control allowed characterization of (1) maximal specific growth rate in exponential growth phases, (2) exponential induction of acid formation in postexponential phases, (3) inhibition of organic acid formation after nerolidol addition, and (4) exponential recovery from this inhibition. Addition of a (+/-)-cis/trans-nerolidol mixture during exponential or postexponential phase to cultures grown in minimal medium at high dissolved oxygen tension (above 50% air saturation), to cultures at low dissolved oxygen tension (5% air saturation), or to cultures grown in rich medium demonstrated that the physiological state before nerolidol addition had a major influence on biotransformation. The maximal molar yield of 12-hydroxy-trans-nerolidol (9%) was obtained by addition of a (+/-)-cis/trans-nerolidol mixture to the culture in the postexponential phase at high dissolved oxygen tension in minimal medium. Similar yields were obtained in rich medium, where the rate of biotransformation was doubled. PMID- 14763866 TI - Kalman filter based glucose control at small set points during fed-batch cultivation of Saccharomyces cerevisiae. AB - A glucose control system is presented, which is able to control cultivations of Saccharomyces cerevisiae even at low glucose concentrations. Glucose concentrations are determined using a special flow injection analysis (FIA) system, which does not require a sampling module. An extended Kalman filter is employed for smoothing the glucose measurements as well as for the prediction of glucose and biomass concentration, the maximum specific growth rate, and the volume of the culture broth. The predicted values are utilized for feedforward/feedback control of the glucose concentration at set points of 0.08 and 0.05 g/L. The controller established well-defined conditions over several hours up to biomass concentrations of 13.5 and 20.7 g/L, respectively. The specific glucose uptake rates at both set points were 1.04 and 0.68 g/g/h, respectively. It is demonstrated that during fed-batch cultivation an overall pure oxidative metabolism of glucose is maintained at the lower set point and a specific ethanol production rate of 0.18 g/g/h at the higher set point. PMID- 14763867 TI - Retention and viability characteristics of mammalian cells in an acoustically driven polymer mesh. AB - A processing approach for the collection and retention of mammalian cells within a high porosity polyester mesh having millimeter-sized pores has been studied. Cell retention occurs via energizing the mesh with a low intensity, resonant acoustic field. The resulting acoustic field induces the interaction of cells with elements of the mesh or with each other and effectively prevents the entrainment of cells in the effluent stream. Experiments involving aqueous suspensions of polystyrene particles were used to provide benchmark data on the performance of the acoustic retention cell. Experiments using mouse hybridoma cells showed that retention densities of over 1.5 x 10(8) cell/mL could be obtained. In addition, the acoustic field was shown to produce a negligible effect on cell viability for short-term exposure. PMID- 14763868 TI - Stabilization of a multimeric beta-galactosidase from Thermus sp. strain T2 by immobilization on novel heterofunctional epoxy supports plus aldehyde-dextran cross-linking. AB - This work exemplifies the advantages of using a battery of new heterofunctional epoxy supports to immobilize enzymes. We have compared the performance of a standard Sepabeads-epoxy support with other Sepabeads-epoxy supports partially modified with boronate, iminodiacetic, metal chelates, and ethylenediamine in the immobilization of the thermostable beta-galactosidase from Thermus sp. strain T2 as a model system. Immobilization yields depended on the support, ranging from 95% using Sepabeads-epoxy-chelate, Sepabeads-epoxy-amino, or Sepabeads-epoxy boronic to 5% using Sepabeads-epoxy-IDA. Moreover, immobilization rates were also very different when using different supports. Remarkably, the immobilized beta galactosidase derivatives showed very improved but different stabilities after favoring multipoint covalent attachment by long-term alkaline incubation, the enzyme immobilized on Sepabeads-epoxy-boronic being the most stable. This derivative had some subunits of the enzyme not covalently attached to the support (detected by SDS-PAGE). This is a problem if the biocatalysts were to be used in food technology. The optimization of the cross-linking with aldehyde-dextran permitted the full stabilization of the quaternary structure of the enzyme. The optimal derivative was very active in lactose hydrolysis even at 70 degrees C (over 1000 IU/g), maintaining its activity after long incubation times under these conditions and with no risk of product contamination with enzyme subunits. PMID- 14763869 TI - Static magnetic fields enhancement of Saccharomyces cerevisae ethanolic fermentation. AB - Magnetic effects induced in ethanolic fermentation by Saccharomyces cerevisiae strain DAUFPE-1012 were studied during a 24 h exposure to 220 mT steady magnetic fields (SMF) at 23 +/- 1 degrees C, produced by NdFeB rod magnets. The magnets were attached diametrically opposed (N to S) to a cylindrical tube reactor. The biomass growth in the reactor culture media (yeast extract + glucose 2%) during 24 h was monitored by measurements of optical density, which was correlated to cell dry weight. Ethanol concentration and glucose level were measured every 2 h. The pH of the culture media was maintained between 4 and 5. As a result, biomass (g/L) increased 2.5-fold and ethanol concentration 3.4-fold in magnetized cultures (n = 8) as compared with SMF nonexposed cultures (n = 8). Glucose consumption was higher in magnetized cultures, which correlated to the ethanol yield. PMID- 14763882 TI - Leprosy in Greece at the end of the 20th century (1988-2000). PMID- 14763870 TI - Allyl-thiosulfinates, the bacteriostatic compounds of garlic against Helicobacter pylori. AB - Allicin and allyl-methyl plus methyl-allyl thiosulfinate from acetonic garlic extracts (AGE) have been isolated by high-performance liquid chromatography. These compounds have shown inhibition of the in vitro growth of Helicobacter pylori (Hp), the bacterium responsible for serious gastric diseases such as ulcers and even gastric cancer. A chromatographic method was optimized and used to isolate these thiosulfinates. The method developed has allowed the isolation of natural thiosulfinates extracted from garlic by organic solvents and is an easy and cheap methodology that avoids complex synthesis and purification procedures. The capacity and effectiveness of isolated natural thiosulfinates have been tested, and this has enabled the identification of the main compounds responsible for the bacteriostatic activity shown by AGE origin of these kinds of organosulfur compounds along with ethanolic garlic extracts (EGE). Additionally, microbiological analyses have suggested that these compounds show a synergic effect on the inhibition of the in vitro growth of Hp. The results described here facilitate the process of obtaining garlic extracts with optimal bacteriostatic properties. The product is obtained in a way that avoids expensive purification methods and will allow the design of live tests with the aim of investigating the potential for the use of these garlic derivatives in the treatment of patients with Hp infections. PMID- 14763883 TI - Incomprehension. PMID- 14763884 TI - Mechanisms involved in peripheral nerve damage in leprosy with special reference to insights obtained from in vitro studies and the experimental mouse model. AB - The histopathological observations of Khanolkar and Iyer, that M. leprae has a predeliction for nerves, first highlighted the central role of peripheral nerves in the pathology of leprosy. It is now well recognized that nerve damage in leprosy will still continue to be an important problem in control and rehabilitation despite the presence of more efficient therapy. The multiplicity of mechanisms postulated, identified, and demonstrated in the last three decades has received little recognition from the scientific community at large. This review is therefore an attempt to collate these multiple studies on mechanisms of nerve damage into a cohesive analysis, which would facilitate the design of future studies. The objective of this review is to focus therefore only on studies which serve to illustrate mechanisms of nerve damage. PMID- 14763885 TI - Leprosy and psoriasis: an enigmatic relationship. AB - The relationship between leprosy and psoriasis has been controversial since ancient times. Based on the fundamental importance of nerve involvement in the pathogenesis of leprosy and psoriasis, it has been hypothesized that leprosy patients may be protected from developing psoriasis. There are only sporadic reports of coexistence of these two diseases as evidence of this negative association. We report a 64-year-old male patient with borderline leprosy and psoriasis. Recent advances in the elucidation of pathogenesis of both diseases have contributed to the understanding of this enigmatic relationship. Various genetic, immunological, and structural alterations in leprosy and psoriasis as discussed could be responsible for the rare co-existence of these two diseases in a given patient. PMID- 14763886 TI - Ultrastructural study of Schwann cells and endothelial cells in the pathogenesis of leprous neuropathy. AB - Peripheral nerve biopsies from 4 borderline tuberculoid (BT) and 4 lepromatous (LL) patients who were on multidrug therapy were investigated by light and electron microscopic studies. The variation of diameters and distribution of myelinated and unmyelinated fibers between BT and LL patients were not significant. This study has shown significant changes in peripheral nerves and endoneural blood vessels. It was revealed that besides Schwann cells (SC), the endothelial cells (EC) of endoneural blood vessels frequently harbor M. leprae. In BT, peripheral nerves in addition to the degenerative changes of SC and presence of perineural and perivascular cuffing by mononuclear cells, the endoneurial blood vessels showed thickening of basement membrane with hypertrophy of EC leading to narrowing or complete occlusion of lumen. On the other hand, peripheral nerves of LL patients were infiltrated with large number of M. leprae shown to be present in the electron transparent zone (ETZ) of the SC. The EC of endoneurial blood vessels were found to be loaded with M. leprae, and this bacillary loaded EC was found to release M. leprae into the lumen through its ruptured membrane. PMID- 14763887 TI - Nail involvement in leprosy: a study of 300 patients. AB - Three hundred leprosy patients were recruited to study the pattern and frequency of nail changes. Nail changes, like longitudinal ridging in finger nails, transverse striations involving both finger and toe nails etc. which occurred with similar frequency in the PB and MB patients in comparison with the control group, were excluded from the analysis. Out of a total number of 150 PB patients, 84 (56%) showed nail changes. Fifty-eight (38.6%) patients showed changes in the finger nails, with an average of 3.2 involved nails per patient. Fifty-three (35.3%) patients showed changes in the toe nails, with an average of 3.0 nails per patient. The most common change observed was longitudinal melanonychia (32.4%) in the finger nails and longitudinal ridging (46.3%) in the toe nails.In comparison, 131/150 (87.3%) MB patients showed nail changes. Finger nail changes were seen in 102 (68%) patients with an average of 5.5 nails affected per patient. Changes in toe nails were seen in 116 (77.3%) patients, with an average of 6.0 nails involved per patient. The most common nail change observed was longitudinal melanonychia in 89/523, (17%) of the total involved finger nails and subungual hyperkeratosis in 164/702, (23.4%) of the total toe nails involvement. Out of a total of 32 colony patients, 31 (96.9%) showed nail changes both in finger and toe nails with an average of 7.9 and 8.4 affected nails per patient, respectively. The most common nail change observed was rudimentary nail(s) on fingers (29%) and toes (21.1%). Among MB patients, a significantly higher number had finger nail involvement in LL group. The frequency of nail involvement for both fingers and toes was significantly greater in LL as compared to BL group of patients. The frequency of nail involvement was significantly more in patients having disease for more than 5 years and in those having trophic changes secondary to loss of sensations and impaired circulation. PMID- 14763888 TI - Long-term efficacy of 2 year WHO multiple drug therapy (MDT) in multibacillary (MB) leprosy patients. AB - Relapse rate estimates after 2 year WHO multiple drug therapy (MDT) in multi bacillary (MB) leprosy vary. Between 1987 and 1994, 500 MB leprosy patients completing 2 year MDT were enrolled in a prospective relapse study. The majority of patients (N = 316) were treated and followed at the physician-staffed Cebu Skin Clinic (CSC), whereas others (N = 184) received therapy from government clinics and were followed by CSC technicians in the field. Relapse definition was an increased bacteriologic index (BI) and new skin lesions, supplemented with mouse footpad inoculations. Through 2002, follow-up was 5368 person-years, with a mean of 10.8 years per patient. The absolute relapse rate was 3% (15/498; 0.28/100 person-years), with a cumulative risk estimate of 3.9% at 15 yrs. For a subset of 217 patients followed for >or=12 yrs or until relapse, relapses occurred in 9% (13/142) attending the CSC, versus 3% (2/75) assessed in the field (p = 0.09). The rate for patients followed at CSC for >or=12 yrs and a pre treatment BI >or=2.7+ was 13% (13/98). All relapses were BL or LL, with pre treatment BI's of >or=2.7+. Relapses occurred long after completion of therapy, between 3 and 11 yrs from the midpoint of the examination without relapse to detection, or between 6 to 13 yrs to the actual year of detection, 7 occurring at >or=10 yrs. Lesion material from all relapses contained M. leprae that was rifampin and clofazimine sensitive, whereas 3 showed partial or full dapsone resistance. [Follow-up rigor and time], medical expertise, and pre-treatment bacterial load influence relapse rates after 2 yr MDT. PMID- 14763898 TI - Can lymphatic drainage be measured non-invasively in human limbs, using plethysmography? AB - There is always rapid volume restitution of the accumulated interstitial fluid after a VCP (venous congestion plethysmography) protocol. It has been suggested that backward extrapolation of the relationship between applied hydrostatic pressure and fluid filtration may give a measure of tissue J (vL) (lymph flow); if so, this could be of immense value in pathophysiological investigations. We hypothesized that the congestion pressure decrease following the VCP protocol might be the stimulus for activating the observed rapid interstitial fluid removal mechanism. We investigated this hypothesis by using a cumulative small step VCP protocol to a maximum arterial diastolic pressure, followed by a mirror image of step pressure decreases. The increases and decreases in cuff pressure produced capillary filtration capacities that were not significantly different from one another [(3.8+/-1.0) x 10(-3) and (3.7+/-1.2) x 10(-3) ml x 100 ml(-1) x min(-1) x mmHg(-1) respectively]. However, we did observe a significant 3-fold increase in estimated lymph flow between the up and 'mirror' down protocol. Moreover, the calculated supine control value, reflecting interstitial fluid removal ( J (vL)), of 0.03+/-0.03 ml x 100 ml(-1) x min(-1) was within the range of lymph flows in human limbs described by other workers, as was the 3-fold increase to 0.09+/-0.03 ml x 100 ml(-1) x min(-1) following the release of the venous congestion. These results support the notion that strain-gauge plethysmography might provide a non-invasive means of assessing peripheral lymph flow in human limbs. PMID- 14763899 TI - Putrescine biosynthesis in mammalian tissues. AB - L-ornithine decarboxylase provides de novo putrescine biosynthesis in mammals. Alternative pathways to generate putrescine that involve ADC (L-arginine decarboxylase) occur in non-mammalian organisms. It has been suggested that an ADC-mediated pathway may generate putrescine via agmatine in mammalian tissues. Published evidence for a mammalian ADC is based on (i) assays using mitochondrial extracts showing production of 14CO2 from [1-14C]arginine and (ii) cloned cDNA sequences that have been claimed to represent ADC. We have reinvestigated this evidence and were unable to find any evidence supporting a mammalian ADC. Mitochondrial extracts prepared from freshly isolated rodent liver and kidney using a metrizamide/Percoll density gradient were assayed for ADC activity using L-[U-14C]-arginine in the presence or absence of arginine metabolic pathway inhibitors. Although 14CO2 was produced in substantial amounts, no labelled agmatine or putrescine was detected. [14C]Agmatine added to liver extracts was not degraded significantly indicating that any agmatine derived from a putative ADC activity was not lost due to further metabolism. Extensive searches of current genome databases using non-mammalian ADC sequences did not identify a viable candidate ADC gene. One of the putative mammalian ADC sequences appears to be derived from bacteria and the other lacks several residues that are essential for decarboxylase activity. These results indicate that 14CO2 release from [1 14C]arginine is not adequate evidence for a mammalian ADC. Although agmatine is a known constituent of mammalian cells, it can be transported from the diet. Therefore L-ornithine decarboxylase remains the only established route for de novo putrescine biosynthesis in mammals. PMID- 14763900 TI - Conversion into GABA (gamma-aminobutyric acid) may reduce the capacity of L glutamine as an insulin secretagogue. AB - We have carried out a detailed examination of L-glutamine metabolism in rat islets in order to elucidate the paradoxical failure of L-glutamine to stimulate insulin secretion. L-Glutamine was converted by isolated islets into GABA (gamma aminobutyric acid), L-aspartate and L-glutamate. Saturation of the intracellular concentrations of all of these amino acids occurred at approx. 10 mmol/l L glutamine, and their half-maximal values were attained at progressively increasing concentrations of L-glutamine (0.3 mmol/l for GABA; 0.5 and 1.0 mmol/l for Asp and Glu respectively). GABA accumulation accounted for most of the 14CO2 produced at various L-[U-14C]glutamine concentrations. Potentiation by L glutamine of L-leucine-induced insulin secretion in perifused islets was suppressed by malonic acid dimethyl ester, was accompanied by a significant decrease in islet GABA accumulation, and was not modified in the presence of GABA receptor antagonists [50 micromol/l saclofen or 10 micromol/l (+)-bicuculline]. L Leucine activated islet glutamate dehydrogenase activity, but had no effect on either glutamate decarboxylase or GABA transaminase activity, in islet homogenates. We conclude that (i) L-glutamine is metabolized preferentially to GABA and L-aspartate, which accumulate in islets, thus preventing its complete oxidation in the Krebs cycle, which accounts for its failure to stimulate insulin secretion; (ii) potentiation by L-glutamine of L-leucine-induced insulin secretion involves increased metabolism of L-glutamate and GABA via the Krebs cycle (glutamate dehydrogenase activation) and the GABA shunt (2-oxoglutarate availability for GABA transaminase) respectively, and (iii) islet release of GABA does not seem to play an important role in the modulation of the islet secretory response to the combination of L-leucine and L-glutamine. PMID- 14763901 TI - A mechanistic insight into a proteasome-independent constitutive inhibitor kappaBalpha (IkappaBalpha) degradation and nuclear factor kappaB (NF-kappaB) activation pathway in WEHI-231 B-cells. AB - Inducible activation of the transcription factor NF-kappaB (nuclear factor kappaB) is classically mediated by proteasomal degradation of its associated inhibitors, IkappaBalpha (inhibitory kappaBalpha) and IkappaBbeta. However, certain B-lymphocytes maintain constitutively nuclear NF-kappaB activity (a p50-c Rel heterodimer) which is resistant to inhibition by proteasome inhibitors. This activity in the WEHI-231 B-cell line is associated with continual and preferential degradation of IkappaBalpha, which is also unaffected by proteasome inhibitors. Pharmacological studies indicated that there was a correlation between inhibition of IkappaBalpha degradation and constitutive p50-c-Rel activity. Domain analysis of IkappaBalpha by deletion mutagenesis demonstrated that an N-terminal 36-amino-acid sequence of IkappaBalpha represented an instability determinant for constitutive degradation. Moreover, domain grafting studies indicated that this sequence was sufficient to cause IkappaBbeta, but not chloramphenicol acetyltransferase, to be rapidly degraded in WEHI-231 B-cells. However, this sequence was insufficient to target IkappaBbeta to the non proteasome degradation pathway, suggesting that there was an additional cis element(s) in IkappaBalpha that was required for complete targeting. Nevertheless, the NF-kappaB pool associated with IkappaBbeta now became constitutively active by virtue of IkappaBbeta instability in these cells. These findings further support the notion that IkappaB instability governs the maintenance of constitutive p50-c-Rel activity in certain B-cells via a unique degradation pathway. PMID- 14763902 TI - Thermodynamic characterization of monomeric and dimeric forms of CcdB (controller of cell division or death B protein). AB - The protein CcdB (controller of cell division or death B) is an F-plasmid-encoded toxin that acts as an inhibitor of Escherichia coli DNA gyrase. The stability and aggregation state of CcdB have been characterized as a function of pH and temperature. Size-exclusion chromatography revealed that the protein is a dimer at pH 7.0, but a monomer at pH 4.0. CD analysis and fluorescence spectroscopy showed that the monomer is well folded, and has similar tertiary structure to the dimer. Hence intersubunit interactions are not required for folding of individual subunits. The stability of both forms was characterized by isothermal denaturant unfolding and calorimetry. The free energies of unfolding were found to be 9.2 kcal x mol(-1) (1 cal approximately 4.184 J) and 21 kcal x mol(-1) at 298 K for the monomer and dimer respectively. The denaturant concentration at which one half of the protein molecules are unfolded (C(m)) of the dimer is dependent on protein concentration, whereas the C(m) of the monomer is independent of protein concentration, as expected. Although thermal unfolding of the protein in aqueous solution is irreversible at neutral pH, it was found that thermal unfolding is reversible in the presence of GdmCl (guanidinium chloride). Differential scanning calorimetry in the presence of low concentrations of GdmCl in combination with isothermal denaturation melts as a function of temperature were used to derive the stability curve for the protein. The value of Delta C (p) (representing the change in excess heat capacity upon protein denaturation) is 2.8+/-0.2 kcal x mol(-1) x K(-1) for unfolding of dimeric CcdB, and only has a weak dependence on denaturant concentration. PMID- 14763903 TI - The identification of a reaction site of glutathione mixed-disulphide formation on gammaS-crystallin in human lens. AB - The glutathionylation of human lens proteins was examined by Western-blot analysis with an anti-GSH antibody and scanning. Several different glutathionylated proteins were observed, and a 47 kDa band was of particular interest. This band did not appear after SDS/PAGE under reducing conditions, suggesting that it was a glutathionylated fraction. The 47 kDa band was found principally in the outer part of the lens, the cortex, but not in the lens nucleus where older proteins are present. The 47 kDa component was composed of betaB1-, betaB2- and gammaS-crystallin, with the gammaS-crystallin having glutathione bound at Cys-82 and at Cys-22, Cys-24 or Cys-26. We conclude that when glutathione becomes bound to gammaS-crystallin, it causes it to bind in turn to the beta-crystallin polypeptides to form a dimer. PMID- 14763904 TI - Production and characterization of hemicellulase activities from Trichoderma harzianum strain T4. AB - Xylan and mannan are the major constituent groups of hemicellulose in the cell wall of higher plants. The mesophilic fungus Trichoderma harzianum strain T4 produces extracellular xylanase and mannanase activities when grown in the presence of oat (Avena sativa)-spelt xylan and wheat bran as the carbon sources respectively. After the growth procedure, the crude extracts were submitted to ultrafiltration in an Amicon system fitted with a 10 kDa-cut-off membrane. Mannanase activity was only detected in the concentrated sample, whereas xylanase was also found in the permeate after ultrafiltration. Xylanase from the concentrated sample showed highest activity at 40 degrees C and pH 5.0. Mannanase activity was optimal at 65 degrees C and pH 2.6. Xylanase was stable in the temperature range 40-70 degrees C, presenting full stability for at least 48 h. Xylanase retained 100% of its original activity after incubation for 48 h at 70 degrees C. Xylanase was also stable at pH 5.0 and 6.0 for 48 h. However, mannanase activity was markedly less stable. The enzyme lost 50% of its activity at 55 degrees C after 45 min, whereas at 60 degrees C its half-life was 20 min. The Michaelis-Menten constant K(m) and V(max) for mannanase and xylanase activities were also calculated. Xylanase had more affinity for soluble xylan, with K(m) and V(max) values of 1.61 mg/ml and 10.03 units/ml respectively. The K(m) and V(max) values for crude mannanase were 6.0 mg/ml and 20.1 units/ml respectively. Xylanase and mannanase were activated by dithiothreitol, L-cysteine and L-tryptophan. Xylanase was partially purified by gel-filtration (Sephadex G 50) and hydrophobic-interaction (Phenyl-Sepharose) chromatographies. The partially purified enzyme was stable over the pH range 5-7 and temperature range of 40-60 degrees C. It was more active on soluble oat-spelt xylan and was activated by dithiothreitol, L-cysteine and L-tryptophan. PMID- 14763905 TI - Importance of quantifying insulin secretion in relation to insulin sensitivity to accurately assess beta cell function in clinical studies. AB - Insulin sensitivity and insulin secretion are mutually related such that insulin resistance is compensated by increased insulin secretion. A correct judgement of insulin secretion therefore requires validation in relation to the insulin sensitivity in the same subject. Mathematical analyses of the relationship between insulin sensitivity and insulin secretion has revealed a hyperbolic function, such that the product of the two variables is constant. This product is usually called the disposition index. Several techniques may be used for its estimation such as data derived from the frequently sampled i.v. glucose tolerance test, the oral glucose tolerance test or the glucose-dependent arginine stimulation test or the euglycemic hyperinsulinemic clamp technique in combination with a test on insulin secretion. Using these techniques the compensatory increase in beta cell function in insulin resistance has been verified in obesity, in pregnancy and after glucocorticoid administration as has the defective beta cell function as the underlying cause of impaired glucose tolerance and type 2 diabetes. Similarly, combined analysis of insulin sensitivity and insulin secretion has shown a down-regulation of beta cell function in increased insulin sensitivity accompanying weight reduction in obesity and following exercise. Acknowledging this inverse relationship between insulin secretion and insulin sensitivity therefore requires estimation of both variables for correct assessment in any individual. PMID- 14763906 TI - Follow-up of low-risk patients with differentiated thyroid carcinoma: a European perspective. AB - OBJECTIVE: Because differentiated (follicular and papillary) thyroid cancer (DTC) may recur years after initial treatment, the follow-up of patients with DTC is long term. However, this population has changed, with more individuals being discovered at an earlier stage of the disease, so that previous follow-up protocols based mostly on data from high-risk patients no longer apply. We sought to develop an improved protocol for the follow-up of low-risk patients with DTC based on the findings of recent studies. METHODS: We analysed recent literature on the follow-up of DTC. RESULTS: Recent large studies have produced three important findings: (i) in patients with low-risk DTC with no evidence of disease up to the 6- to 12-month follow-up, diagnostic whole-body scan adds no information when serum thyroglobulin (Tg) is undetectable and interference from anti-Tg antibodies is absent; (ii) use of recombinant human thyroid-stimulating hormone to aid Tg measurement is effective and provides greater safety, quality of-life and work productivity than does levothyroxine withdrawal with its attendant hypothyroidism; and (iii) ultrasonography performed by an experienced operator is the most sensitive means of detecting neck recurrences of DTC. CONCLUSIONS: We present a revised follow-up protocol for low-risk patients taking into account the above findings. This protocol should help clinicians enter a new era of monitoring characterized by greater safety, simplicity, convenience and cost savings. PMID- 14763907 TI - Estrogens and atherosclerosis. AB - Numerous epidemiological as well as experimental studies have suggested that estradiol (E2) prevents atherosclerosis development. However two controlled prospective and randomized studies in women using hormone replacement therapy (HRT) did not confirm this beneficial effect. We then decided to use mouse models of atherosclerosis to define the possible mechanisms involved and the reasons for the discrepancy. We have shown that, although serum cholesterol decreases, this influence on lipid metabolism is negligible. Surprisingly, E2 induces an inflammatory-immune response towards a T helper cell (Th1) profile with increasing interferon-gamma production that could destabilize atheromatous plaques, and could account for the increase in the frequency of cardiovascular events in women undergoing HRT. At the level of the endothelium, E2 induces an increase in nitric oxide (NO) biodisponibility, but this phenomenon does not concern the development of fatty streaks. Nevertheless, the atheroprotective effect is apparently mediated at the level of the endothelium by a mechanism that has still to be characterized in molecular terms. These new acquisitions constitute a basis for new pharmacological developments allowing the prevention of deleterious effects and preserving the beneficial ones. PMID- 14763908 TI - Precocious puberty secondary to massive ovarian oedema in a 6-month-old girl. AB - A 6-month-old girl was referred with breast and pubic hair development. Investigations excluded an adrenal or central cause for her precocity. Ovarian ultrasound scans showed bilaterally enlarged ovaries with both solid and cystic changes. A follow-up examination suggested progression of the precocity and in view of the young age of the child, and concerns regarding underlying malignancy, she underwent laparotomy. Histology showed no evidence of neoplasia but there was stromal oedema consistent with a diagnosis of massive ovarian oedema. This entity is poorly recognised in the paediatric literature as a cause of sexual precocity, and has never previously been described in such a young patient. This is an unusual cause of precocity in a young child and its recognition and management are reviewed. PMID- 14763909 TI - Change in von Willebrand factor and carotid intima-media thickness in hypothyroid patients with normal thyroid function after levothyroxine replacement therapy. AB - OBJECTIVE: This study examined whether levothyroxine (L-T4) replacement might affect serum markers of endothelium injury, including von Willebrand factor (vWF), factor VIII activity and thrombomodulin (TM), during normalization of increased intima-media thickness (IMT) in the common carotid artery (CCA) in hypothyroid patients after L-T4 replacement therapy. PATIENTS AND METHODS: Thirty three hypothyroid patients were examined for vWF, factor VIII, TM and CCA IMT before and after 1 year of normalization of thyroid function by L-T4 replacement. CCA IMT was measured from digitized still images taken during scanning by high resolution ultrasonography as an indicator of early atherosclerosis. RESULTS: Serum factor VIII and vWF increased significantly during 1 year of normalization of thyroid function (from 122.7+/-9.4 to 151.3+/-18.8% (P<0.05) and from 109.9+/ 9.6 to 135.2+/-12.4% (P<0.005) respectively), although these values all fell within the respective normal range. Serum TM, in contrast, did not change appreciably in response to L-T4 treatment, moving from 2.57+/-0.15 to 2.74+/-0.18 ng/ml (P=0.086). During 1 year of a euthyroid state, all patients showed a significant decrease in CCA IMT (P<0.0001). Change in serum vWF, but not in factor VIII or TM, showed a positive correlation with that of CCA IMT during L-T4 replacement therapy. Furthermore, the change in serum vWF was significantly and independently associated with change in CCA IMT (r=0.490, P=0.0038). CONCLUSIONS: The present study demonstrated that the improvement of CCA IMT during L-T4 treatment might have the potential to attenuate an elevation of vWF and to attenuate vascular injury by the cardiovascular effects of thyroid hormone in hypothyroid patients. PMID- 14763910 TI - Advances in diagnostic practices affect thyroid cancer incidence in France. AB - OBJECTIVE: To analyse trends in diagnostic practices of thyroid diseases and to relate them to the increase in thyroid cancer incidence in France over time. DESIGN: From 1980 to 2000, a French retrospective multicentric (three endocrinology and three nuclear medicine centres) study of thyroid diseases was conducted on 20 consecutive unselected patients' records, sampled every 5 years in each centre. METHODS: Characteristics of the population and diagnosis procedures (thyroid ultrasonography (US), radionuclide scan, cytology and hormonal measurements) were described over time. Changing trends in operated patients and in cancer prevalence were analysed as well as the impact of practices on cancer incidence. RESULTS: The study included 471 patients (82% female, mean age 46.7, range 9-84 years), referred for nodular thyroid diseases (66.7%) or thyroid dysfunctions (33.3%). A significant increase in US (3 to 84.8%) and cytological practices (4.5 to 23%), and a decrease (89.4 to 49.6%) in radionuclide scan procedures were observed over time. Although the proportion of patients undergoing surgery remained constant (24.8%), the prevalence of cancer increased among operated patients from 12.5 to 37% (P=0.006). In a Cox's proportional hazard model stratified on the clinical characteristics of patients, only the cytological practice, regardless of its results, was significantly associated with the occurrence of cancer: relative risk (RR)=4.4 (95% confidence interval (CI): 1.1-16; P=0.04). CONCLUSIONS: From 1980 to 2000, a major evolution in clinical practices has led to the increase in thyroid cancer reported in France. Such changes in medical, as well as in surgical and pathological, practices must be taken into account in incidence measurement. PMID- 14763911 TI - Video-assisted thoracoscopic surgery as a first-line treatment for mediastinal parathyroid adenomas: strategic value of imaging. AB - OBJECTIVE: To present first-line thoracic surgery made possible by localization studies in three patients with ectopic parathyroid adenomas. DESIGN AND METHODS: Three patients with ectopic parathyroid tissue in the mediastinum were examined by ultrasound, technetium-99m sestamibi scintigraphy, computed tomography (CT), and venous catheterization with measurement of parathyroid hormone. Without previous cervical exploration, video-assisted thoracic surgery (VATS) was used in all cases to avoid the need for thoracic open surgical procedures. RESULTS AND CONCLUSIONS: The mediastinal parathyroid glands were all detected at scintigraphy, and CT and venous catheterization were helpful in anatomic and functioning characterization. All pathologic glands were successfully resected, with only one minor complication. VATS can safely remove a deep mediastinal parathyroid adenoma and avoid more aggressive open approaches. In an experienced referral center, systematic and sophisticated imaging studies may accurately identify and localize rare ectopic parathyroid adenomas, and avoid cervical surgery. PMID- 14763912 TI - Direct causes of death in Japanese patients with hypopituitarism as analyzed from a nation-wide autopsy database. AB - OBJECTIVES: The direct causes of death in Japanese patients with hypopituitarism remain unclear. In this study, the direct causes of death were investigated and compared between Japanese patients with hypopituitarism from a nation-wide autopsy database and an age- and gender-matched control population from national reports. SUBJECTS: Three hundred and ninety-one subjects with hypopituitarism who had died were selected from a nation-wide autopsy database (1984-1993). The ratios of each cause of death among the age- and gender-matched control population were derived from national reports. RESULTS: In subjects with hypopituitarism, an increased relative frequency of death from cerebrovascular diseases (male; 2.02 (95% confidence interval (CI) 1.45-2.82), female; 1.73 (95% CI 1.18-2.52)) was found. In particular, the relative frequency of death from cerebral hemorrhage was 4.60 (95% CI 2.95-7.17) in male and 4.80 (95% CI 2.90 7.94) in female subjects with hypopituitarism. Unexpectedly, a decreased relative frequency of death from all heart diseases (male; 0.439 (95% CI 0.277-0.696), female; 0.267 (95% CI 0.149-0.478)) was found in subjects with hypopituitarism, although there was no difference between subjects with hypopituitarism and controls in the frequency of death from ischemic heart disease. CONCLUSIONS: These results provide useful information for the long-term care of Japanese patients with hypopituitarism. PMID- 14763913 TI - Utility of P300 auditory event related potential latency in detecting cognitive dysfunction in growth hormone (GH) deficient patients with Sheehan's syndrome and effects of GH replacement therapy. AB - OBJECTIVE: Impaired cognitive function has been demonstrated in adults with growth hormone (GH) deficiency (GHD) by using different neuropsychological tests. Despite several studies, present knowledge about the impact of GHD and GH replacement therapy (GHRT) on cognitive function is limited. P300 event-related potential (ERP) application is a well-established neurophysiological approach in the assessment of cognitive functions including the updating of working memory content and the speed of stimulus evaluation. GHD is a well-known feature of Sheehan's syndrome and cognitive changes due to GHD and the effects of GHRT remain to be clarified. The present study was designed to investigate the effects of GHD and 6 months of GHRT on cognitive function in patients with Sheehan's syndrome by using P300 latency. DESIGN AND METHODS: The study comprised 14 patients with Sheehan's syndrome (mean age, 49.5+/-7.8 years) and 10 age-, education- and sex-matched healthy controls. With hormone replacement therapy, basal hormone levels other than GH were stable before enrollment and throughout the GHRT. The diagnosis of GH deficiency was established by insulin-tolerance test (ITT), and mean peak level of GH in response to insulin hypoglycemia was 0.77+/-0.35 mIU/l. Treatment with GH was started at a dose of 0.45 IU (0.15 mg)/day in month 1, was increased to 0.9 IU (0.30 mg)/day in month 2 and was maintained at 2 IU (0.66 mg)/day. Initially baseline auditory ERPs in patients and controls were recorded at frontal (Fz), central (Cz), and parietal (P3 and P4) electrode sites. In the patient group, ERPs were re-evaluated after 6 months of GH replacement therapy. During each session P300 amplitude and latency were measured. RESULTS: Mean serum insulin-like growth factor-I (IGF-I) concentration in the patient group before GHRT was 23+/-13 ng/ml. After 6 months of GH therapy mean IGF-I significantly increased to an acceptable level, 234+/-71 ng/ml (P<0.05). The mean latencies (at all electrode sites) of the patients before GHRT were found to be significantly prolonged when compared with those of normal controls (P<0.05). After 6 months of GHRT mean P300 latencies (at all electrode sites) were decreased significantly when compared with latencies before treatment (P<0.05). CONCLUSIONS: The present study, using P300 ERP latencies, therefore suggests an impairment of cognitive abilities due to severe GHD in patients with Sheehan's syndrome and an improvement of cognitive function after 6 months of physiological GHRT. Moreover, this was a novel application of P300 ERP latencies in cognitive function detection in patients with GHD. Further studies with different patient groups need to be done to assess the clinical use of this electrophysiological method in the diagnosis of cognitive dysfunction due to GHD. PMID- 14763915 TI - Expression of ghrelin and biological activity of specific receptors for ghrelin and des-acyl ghrelin in human prostate neoplasms and related cell lines. AB - BACKGROUND: Ghrelin, a natural growth hormone secretagogue (GHS), has been identified in prostate carcinoma cell lines. OBJECTIVES: To investigate the presence of ghrelin and its receptors in human prostate tumours and in DU-145, PC 3 and LNCaP prostate carcinoma cell lines, and to assess the effects of ghrelin and its more abundant circulating form, des-octanoyl ghrelin, on cell proliferation. METHODS: Ghrelin and types 1a and 1b GHS receptor (GHS-R) were determined at the mRNA and protein levels by RT-PCR, in situ hybridization, immunohistochemistry and enzyme immunoassay in tissues, cell lines and culture medium. Ghrelin binding was determined by radioreceptor assay. The effects on cell proliferation were evaluated by growth curves. RESULTS: Ghrelin mRNA was found in prostatic carcinomas and benign hyperplasias, but immunohistochemistry was negative. GHS-R1a and 1b mRNAs were absent from carcinomas, but GHS-R1b mRNA was present in 50% of hyperplasias. Ghrelin peptide and mRNA were present in PC-3 cells exclusively, whereas GHS-R1a and 1b mRNAs were expressed in DU-145 cells only. Specific [125I]Tyr4-ghrelin binding was detected in prostate tumour, DU-145 and PC-3 cell membranes and the binding was displaced by ghrelin, synthetic GHS and des-octanoyl ghrelin, which is devoid of GHS-R1a binding affinity and GH releasing activity. Ghrelin and des-acyl ghrelin inhibited DU-145 cell proliferation, displayed a biphasic effect in PC-3 cells and were ineffective in LNCaP cells. CONCLUSIONS: Specific GHS binding sites, other than GHS-R1a and 1b, are present in human prostatic neoplasms. Ghrelin, in addition to des-acyl ghrelin, exerts different effects on cell proliferation in prostate carcinoma cell lines. PMID- 14763916 TI - Influence of short-term dietary weight loss on cortisol secretion and metabolism in obese men. AB - OBJECTIVES: Obesity is associated with increased inactivation of cortisol by hepatic A-ring 5alpha- and 5beta-reductases, impaired hepatic regeneration of cortisol from cortisone by 11beta-hydroxysteroid dehydrogenase type 1 (11HSD1), but increased subcutaneous adipose 11HSD1 activity enhancing local cortisol levels in fat. Cause and effect between obesity and abnormal cortisol metabolism is untested. DESIGN: Acute weight loss was induced by very low calorie diet (VLCD) or starvation in obese men. METHODS: Otherwise healthy males (aged 20-55 years; body mass index (BMI) 30-40 kg/m2) were studied after 6 days on a weight maintenance diet; then after either 6 days of starvation (n=6) or 3 weeks of VLCD (2.55 MJ; n=6); then after 1 week of weight maintenance; and finally after 2 weeks of being allowed to feed ad libitum. Plasma samples were obtained from indwelling cannulae at 0930 h and 1815 h and a 24 h urine collection was completed for analysis of cortisol metabolites by gas chromatography/mass spectrometry. RESULTS: Data are mean+/-S.E.M. BMI fell (kg/m3) from 34.8+/-0.8 at baseline to 31.8+/-1.4 on VLCD and 32.7+/-1.1 on starvation. Starvation caused a rise in plasma cortisol (at 0930 h from 143+/-17 to 216+/-11 nM, P<0.001) but no change in total urinary cortisol metabolites. VLCD did not alter plasma cortisol and markedly reduced cortisol metabolite excretion (from 15.8+/-1.1 mg/day at baseline to 7.0+/-1.1 mg/day, P<0.001). Relative excretion of 5alpha-reduced cortisol metabolites fell on both diets, but there were no changes in cortisol/cortisone metabolite ratios reflecting 11HSD activities. CONCLUSIONS: Weight loss with VLCD in obesity reverses up-regulation of hepatic A-ring reductases and normalises cortisol production rate; in contrast, starvation produces acute stress and further activation of cortisol secretion. We suggest that activation of cortisol secretion is not an irreversible intrinsic abnormality in obese patients, and speculate that dietary content has an important influence on the neuroendocrine response to weight loss. PMID- 14763914 TI - Body mass index, circulating levels of sex-steroid hormones, IGF-I and IGF binding protein-3: a cross-sectional study in healthy women. AB - OBJECTIVE: Excess weight has been associated with increased risk of cancer at several organ sites. In part, this effect may be modulated through alterations in the metabolism of sex steroids and IGF-I related peptides. The objectives of the study were to examine the association of body mass index (BMI) with circulating androgens (testosterone, androstenedione and dehydroepiandrosterone sulfate (DHEAS)), estrogens (estrone and estradiol), sex hormone-binding globulin (SHBG), IGF-I and IGF-binding protein (IGFBP)-3, and the relationship between sex steroids, IGF-I and IGFBP-3. DESIGN AND METHODS: A cross-sectional analysis was performed using hormonal and questionnaire data of 620 healthy women (177 pre- and 443 post-menopausal). The laboratory measurements of the hormones of interest were available from two previous case-control studies on endogenous hormones and cancer risk. RESULTS: In the pre-menopausal group, BMI was not related to androgens and IGF-I. In the post-menopausal group, estrogens, testosterone and androstenedione increased with increasing BMI. The association with IGF-I was non linear, with the highest mean concentrations observed in women with BMI between 24 and 25. In both pre- and post-menopausal subjects, IGFBP-3 did not vary across BMI categories and SHBG decreased with increasing BMI. As for the correlations between peptide and steroid hormones, in the post-menopausal group, IGF-I was positively related to androgens, inversely correlated with SHBG, and not correlated with estrogens. In the pre-menopausal group, similar but weaker correlations between IGF-I and androgens were observed. CONCLUSIONS: These observations offer evidence that obesity may influence the levels of endogenous sex-steroid and IGF-related hormones in the circulation, especially after menopause. Circulating IGF-I, androgens and SHBG appear to be related to each other in post-menopausal women. PMID- 14763917 TI - Body weight and glucose metabolism have a different effect on circulating levels of ICAM-1, E-selectin, and endothelin-1 in humans. AB - BACKGROUND: Endothelial dysfunction and inflammation are present in both type 2 diabetes mellitus (T2DM) and obesity. In this paper we compared the role of weight loss and of glycaemic control in determining circulating levels of ICAM-1, endothelin-1 (ET-1), and E-selectin in patients with morbid (grade 3) obesity. METHODS AND RESULTS: ICAM-1, E-selectin, and ET-1 were higher in obese patients (n=96) than in lean controls (n=30); among obese patients, the three molecules were higher in T2DM patients (n=26) than in patients with normal (NGT, n=43) or impaired (IGT, n=27) glucose tolerance. Sixty-eight obese patients had a significant weight loss induced by bariatric surgery, and showed a significant decrease in blood glucose, HbA1c and all molecules, so that ICAM-1, E-selectin, and ET-1 were not different in NGT, IGT and T2DM patients, and in lean controls; in 13 patients with a small weight loss induced by diet, changes were not significant, in spite of a significant reduction in blood glucose and HbA1c. At stepwise regression, changes in ICAM-1, ET-1, and E-selectin significantly correlated only with change in body mass index. CONCLUSIONS: These data indicate that weight loss is more important than glycaemic control in regulating circulating levels of ICAM-1, ET-1, E-selectin in morbidly obese subjects. PMID- 14763918 TI - Adiponectin is independently associated with glycosylated haemoglobin. AB - BACKGROUND: In humans, adiponectin has been demonstrated to circulate in inverse proportion to the degree of insulin resistance. OBJECTIVE: To investigate the association between adiponectin and glycosylated haemoglobin (HbA1c) in a population-based study. DESIGN AND METHODS: Two hundred and ninety-seven individuals aged 30-75 years were enrolled in a cross-sectional study. They included patients with type 2 (non-insulin-dependent) diabetes mellitus and stable, good metabolic control (n=32) and individuals with glucose intolerance (n=54). Adiponectin was measured using a sandwich enzyme-linked immunosorbent assay (intra-assay and interassay coefficients of variation 3.3 and 7.4% respectively). RESULTS: Adiponectin correlated with age (r=0.161; P=0.006), body mass index (r=-0.197; P=0.001), diastolic blood pressure (r=-0.181; P=0.005), fasting glucose and HbA1c (r=-0.251 and r=-0.22 respectively; P<0.0001), high density lipoprotein cholesterol (r=0.442; P<0.001) and serum triglycerides (r= 362; P<0.001). In multiple regression analysis, sex, age, fasting and post-load glucose, and adiponectin independently contributed to 40% of the variance in HbA1c. Among individuals with normal glucose tolerance, fasting glucose (P=0.0033), post-load glucose (P=0.0015), age (P=0.001) and adiponectin (P=0.0083) independently contributed to 21% of the variance in HbA1c. CONCLUSION: Adiponectin is significantly associated with altered glucose metabolism and independently contributes to the variance of HbA1c in a population-based manner. PMID- 14763919 TI - Evidence of increased visceral obesity and reduced physical fitness in healthy insulin-resistant first-degree relatives of type 2 diabetic patients. AB - OBJECTIVE: First-degree relatives (FDR) of type 2 diabetic patients are often insulin resistant. Visceral obesity is closely linked to both insulin resistance and type 2 diabetes. We therefore hypothesized that the inheritance of an increased tendency to store fat in visceral fat depots may be a characteristic phenotypic feature in FDR contributing to their insulin resistance. DESIGN AND METHODS: We measured fat distribution in 20 FDR and 14 age-, gender- and body mass index (BMI)-matched controls employing dual energy X-ray absorbtiometry (DEXA)- and computed tomography (CT)-scanning. Insulin-stimulated glucose uptake (ISGU) was determined by a hyperinsulinemic clamp and maximal aerobic work capacity (VO2 max) by a bicycle ergometer test. Baseline lipolysis was measured using [3H]palmitate. The activity level of the hypothalamic-pituitary-adrenal axis was assessed as the 24 h urinary (u)-cortisol/creatinine ratio. RESULTS: All subjects had a normal oral glucose tolerance test (OGTT), but FDR were insulin resistant (ISGU: 6.64+/-0.48 vs 9.12+/-0.98 mg/kg ffm/min, P=0.01). Despite similar BMI (25.2+/-0.5 vs 24.8+/-0.7 kg/m2, P=0.61) and overall fat mass (26.4+/ 1.6 vs 24.2+/-2.1%, P=0.41) in FDR vs controls, the amount of visceral adipose tissue was substantially increased (65.9+/-10.0 vs 40.1+/-11.3 cm2, P<0.05) and VO2 max was reduced (52.2+/-3.1 vs 63.3+/-3.9 ml/kg ffm/min, P<0.05) in FDR. Visceral adiposity was inversely correlated with ISGU (FDR: r=-0.52, P<0.05; controls: r=-0.65, P<0.01) and in multiple regression analysis visceral adiposity (P<0.01), VO2 max (P<0.001) and a family history of type 2 diabetes (P<0.05) (r2=0.64) all significantly and independently contributed to the level of ISGU. Baseline palmitate appearance (145+/-10 vs 139+/-15 micromol/min, P=0.74) and the 24 h u-cortisol/creatinine ratio ((24.9+/-1.3 vs 27.4+/-2.0).10(-6), P=0.28) were both comparable in the two groups. CONCLUSION: Healthy but insulin-resistant FDR have enhanced visceral obesity and reduced VO2 max compared with people without a family history of diabetes, despite similar BMI and overall fat mass. Both the visceral adiposity and reduced aerobic fitness are strongly associated with and may contribute to their insulin resistance. PMID- 14763920 TI - Metabolic and endocrine effects of treatment with peroral or transdermal oestrogens in conjunction with peroral cyproterone acetate in women with polycystic ovary syndrome. AB - OBJECTIVE: To compare the influence of transdermal and peroral oestrogen treatments in conjunction with cyproterone acetate (CPA) on metabolic and hormonal parameters in women with polycystic ovary syndrome (PCOS). PATIENTS AND METHODS: Twenty-four women with PCOS, aged 25.4+/-4.3 (mean+/-s.d.) years, body mass index 24.5+/-3.9 kg/m2 were randomly assigned to receive either transdermal oestradiol plus CPA (n=12) or a peroral oestradiol-CPA combination (n=12). Before and after 3 months of treatment, basal blood samples, euglycaemic hyperinsulinaemic clamp combined with indirect calorimetry and arginine tests were performed. ANOVA and Student's t-test or Wilcoxon's test were used for statistical analyses. RESULTS: After peroral oestradiol-CPA, insulin sensitivity (P<0.004) and the disposition index as the function of insulin sensitivity and secretion (P<0.0001) decreased significantly. Fasting insulin (P<0.05), cholesterol (P<0.05), high-density lipoprotein cholesterol (P<0.05) and sex hormone binding globulin (P<0.0001) increased significantly. Dehydroepiandrosterone (P<0.05) and 17-OH progesterone (P<0.01) decreased significantly. After transdermal oestradiol+CPA, no significant changes were observed in sex-hormone binding globulin and androgen concentrations, insulin sensitivity or disposition index. CONCLUSIONS: In women with PCOS, peroral oestrogens (at doses common in combined oral contraceptives) led to a significant impairment in insulin secretion and action. In contrast, the transdermal application of oestrogens did not significantly influence insulin sensitivity. PMID- 14763921 TI - The role of CXCR5 and its ligand CXCL13 in the compartmentalization of lymphocytes in thyroids affected by autoimmune thyroid diseases. AB - OBJECTIVE: Graves' disease (GD) and Hashimoto's thyroiditis (HT) are characterized by lymphocytic infiltrates partly resembling secondary lymphoid follicles in the thyroid. CXCR5 and its ligand CXCL13 regulate compartmentalization of B- and T-cells in secondary lymphoid organs. The aim of the study was to elucidate the role of this chemokine receptor-ligand pair in thyroid autoimmunity. METHODS: Peripheral blood and thyroid-derived lymphocyte subpopulations were examined by flow cytometry for CXCR5. CXCR5 and CXCL13 cDNA were quantified in thyroid tissues by real-time RT-PCR. RESULTS: We found no differences between the percentages of peripheral blood CXCR5+ T- and B-cells in GD patients (n=10) and healthy controls (n=10). In GD patients, the number of memory CD4+ cells expressing CXCR5 which are functionally characterized as follicular B helper T-cells is higher in thyroid-derived (18+/-3%) compared with peripheral blood T-lymphocytes (8+/-2%). The highest CXCL13 mRNA levels were found in HT (n=2, 86.1+/-1.2 zmol (10(-21) mol) cDNA/PCR) followed by GD tissues (n=16, 9.6+/-3.5). Only low amounts were determined in thyroid autonomy (TA) (n=11) thyroid tissues, irrespective of whether the autonomous nodule (0.5+/-0.1) or the surrounding normal tissue (1.8+/-0.7) had been analyzed. The same differences were found for CXCR5 (HT: 179.1+/-6.8; GD: 17.4+/-10.6; TA(nodule): 0.8+/-0.5; TA(normal): 4.4+/-3.6). In GD, there is a correlation between CXCL13 and CXCR5 mRNA levels and the number of focal lymphocytic infiltrates and germinal centers as well as anti-thyroperoxidase but not anti-TSH receptor autoantibodies. CONCLUSIONS: CXCR5 and CXCL13 play an essential role in maintaining B- and T-cells in lymphocytic infiltrates and ectopic follicles in thyroid tissue from patients affected by autoimmunity. PMID- 14763922 TI - Effect of GHRH and GHRP-2 treatment in vitro on GH secretion and levels of GH, pituitary transcription factor-1, GHRH-receptor, GH-secretagogue-receptor and somatostatin receptor mRNAs in ovine pituitary cells. AB - OBJECTIVE: Growth hormone (GH)-releasing hormone (GHRH) and GH-releasing peptides (GHRPs) stimulate the release of GH through their specific receptors on somatotropes. Combined GHRH and GHRP administration causes a synergistic GH release in vivo by an unknown mechanism. The current study focuses on the direct action of GHRH and GHRP on several molecular targets in somatotropes. DESIGN AND METHODS: To clarify the mechanism of action, ovine somatotropes were used to measure the expression of mRNAs encoding for GH, pituitary transcription factor-1 (Pit-1), GH-secretagogue receptor (GHS-R), GHRH-R, somatostatin receptor subtypes (sst-1 and sst-2) and GH release after GHRH and GHRP-2 treatment for 0.5, 1, 1.5 and 2 h. RESULTS: GHRH (10 nM), GHRP-2 (100 nM) and combined GHRH-GHRP-2 increased the levels of GH mRNA and GH release from 0.5 to 2 h in a time dependent manner. The levels of Pit-1, GHRH-R and GHS-R mRNA were increased after 0.5 h treatment of cells with GHRH and GHRP-2. The levels of sst-1 but not sst-2 mRNA were significantly increased after 0.5 and 1 h of GHRH treatment. In contrast, both sst-1 and sst-2 mRNA expression was inhibited after 0.5-2 h of GHRP treatment. CONCLUSIONS: These data demonstrate a direct in vitro modification of ovine somatotropes by GHRH and GHRP-2 resulting in altered GHRH R, GHS-R, Pit-1, sst-1, sst-2 and GH gene expression; this may underlie the regulatory action of GHRH and GHRP-2 on GH secretion. PMID- 14763923 TI - The causes of asthma: the need to look at the data with different eyes. PMID- 14763924 TI - Asthma and atopy - the price of affluence? AB - Irrespective of improved knowledge of many aspects of atopic diseases, the unfavorable trends in their prevalence particularly among children could not have been reversed. A growing body of evidence suggests that something may lack from our societal affluence that has the capacity to provide protection against the development of atopic diseases. Much attention during the last years has been devoted to the hygiene hypothesis. This review outlines the impact of environment and lifestyle, particularly from the perspective of the East-West gradient, on the development of atopic diseases, with a special emphasis on the hygiene hypothesis in its broadest sense. PMID- 14763925 TI - Epigenetic inheritance of fetal genes in allergic asthma. AB - Asthma has been associated with an exaggerated T-helper type 2 (Th2) over Th1 responses to allergic and nonallergic stimuli, which leads to chronic airway inflammation and airway remodeling. In the present article, we propose that many of the genes involved in IgE synthesis and airways (re)modeling in asthma are persistent or reminiscent fetal genes which may not be silenced during early infancy (or late pregnancy). Genes of the embryologic differentiation of ectodermic and endodermic tissues may explain some of the patterns of airway remodeling in asthma. In utero programming leads to gene expression, the persistence of which may be associated with epigenetic inheritance phenomena induced by nonspecific environmental factors. Clear delineation of these issues may yield new information on the mechanisms of asthma and new targets for therapeutic intervention and primary prevention. PMID- 14763926 TI - Is the increase in allergic asthma associated with an inborn Th1 maturation or with an environmental Th1 trigger defect? AB - The main reason for the asthma epidemic in industrialized countries is unknown. While childhood mortality from acute respiratory infection is still high in developing countries where asthma prevalence is low, there might be a suppressed natural selection in industrialized countries with a high asthma prevalence. Children with an inborn Th1 maturation defect might survive by better health care and antibiotic use at the cost of higher asthma and allergy rates. Another distinct group of children maybe represented by those having an environmental Th1 trigger deficit where the immune systems is not being sufficiently exposed in early life. Both, a Th1 maturation and a Th1 trigger defect may contribute to a dual Th1 allergy theory. PMID- 14763927 TI - Can we prevent allergy? AB - Allergic conditions continue to increase steeply. The last two decades have seen many prevention trials, studying the effect of dietary and environmental interventions. These trials have yielded invaluable information about the atopic march and also highlighted the need for a clear and commonly used nomenclature as well as a need for better outcome measures. This review discusses primary and secondary prevention studies and their results. PMID- 14763928 TI - Anti-cardiolipin antibodies in atopic dermatitis. PMID- 14763929 TI - Association between the occurrence of the anticardiolipin IgM and mite allergen specific IgE antibodies in children with extrinsic type of atopic eczema/dermatitis syndrome. AB - BACKGROUND: As the literature has only controversial data on the role of nonallergen-specific antibodies in atopic eczema dermatitis syndrome, the authors investigated the link between the occurrence of the antiphospholipid [anticardiolipin (ACL), anti-beta2-glycoprotein I] and allergen-specific immunoglobulin E (IgE) antibodies in 72 children with atopic eczema/dermatitis syndrome (AEDS). METHODS: The measurement of antiphospholipid antibodies was carried out by enzyme-linked immunosorbent assay (ELISA), serum total IgE by nephelometry, and allergen-specific IgE by immunoblotting assay. The statistical analysis was carried out by Fisher's exact test and odds ratio was calculated. RESULTS: Thirteen of 72 children with AEDS (mean age 8.3 years) had elevated serum levels of ACL, and eight anti-beta2-glycoprotein I antibodies. The presence of allergen-specific IgE against inhalant allergens and nutritive allergens was among eight of 13 and three of eight in the cases with elevated ACL. The ratio of patients with highly increased severity scoring of atopic dermatitis (SCORAD) index (>75) was significantly higher in the group with elevated (4/13) than in those with the normal ACL levels (2/59). There was a significant association between the appearance of mite (Dermatophagoides pteronyssinus, D. farinae) specific IgE and ACL IgM antibodies (6/13). CONCLUSION: These findings show that there are significant linkage and association between the appearance of ACL IgM or the production of allergen-specific IgE against inhalant (mainly mite) allergens in children with atopic eczema/dermatitis syndrome. PMID- 14763930 TI - IgE-binding components of cultured human keratinocytes in atopic eczema/dermatitis syndrome and their crossreactivity with Malassezia furfur. AB - BACKGROUND: Atopic eczema/dermatitis syndrome (AEDS) patients display immunoglobulin E (IgE) reactivity to several antigens, e.g. saprophytic yeasts as Malassezia furfur. AEDS patients also show IgE autoreactivity towards cells of their own tissue including epidermis. PURPOSE OF THE STUDY: The aim of this study was to investigate the IgE autoreactivity of AEDS patients to cultured keratinocytes and to reveal potential crossreacting epitopes in cultured keratinocytes and M. furfur. MATERIAL AND METHODS: Serum samples of 27 AEDS patients were analyzed, of these 13 were M. furfur radioallergosorbent test (RAST) positive and 14 negative. Four urticaria, three psoriasis, and seven nonatopic patients were included as controls. The studies were performed by using IgE immunoblotting and immunoblotting inhibition methods. RESULTS: Ten IgE binding protein bands were detected in cultured human keratinocytes by IgE immunoblotting using sera from adult AEDS patients. Anti-keratinocyte IgE antibodies were more associated with elevated S-IgE level than M. furfur RAST. Clear crossreactivity with M. furfur could not be shown. CONCLUSIONS: The possible pathomechanism of anti-keratinocyte IgE antibodies is not due to IgE epitope mimicry of saprophytic yeast and local tissue in AEDS skin. PMID- 14763931 TI - Allergenic pollen records (15 years) and sensitization in patients with respiratory allergy in Thessaloniki, Greece. AB - BACKGROUND: Very limited allergenic pollen records exist in Greece so far; moreover, there is a lack of investigation on patient sensitization. The above data are necessary for respiratory allergy diagnosis and treatment worldwide. METHODOLOGY: Daily records and identification of 16 airborne pollen species were made using a Burkard trap (1987-2001). Skin sensitivity to 13 most common pollen extracts was investigated, in a sample of 1311 asthmatics with atopy, admitted to the Out-Patient Clinic for Asthma (1990-2001). Skin sensitivity to 55 allergens, including 13 pollen extracts, was detected by skin prick test. RESULTS: The following pollen concentrations were recorded: cypress (24.9% of the total), oak (20.8%), wall pellitory (13.6%), olive (9.1%), pine (8.9%), grasses (6.3%), plane (5.4%), hazel (3%), goosefoot (2.5%) and poplar (1.4%). The respective percentages of birch, ragweed, mugwort, willow, alder and elm were lower than 1%. The highest counts of airborne pollen grains were detected from March to June. Regarding patient sensitization, sensitivity was detected to: grasses in 530 patients (40.4%), olive 417 (31.8%), goosefoot 240 (18.3%), wall pellitory 201 (15.3%), mugwort 198 (15.1%), plantain 194 (14.6%), cypress 166 (12.7%), hazel 126 (9.6%), pine 122 (9.3%), poplar 111 (8.4%), plane 107 (8,2%), oak 99 (7.6%) and to birch 89 patients (6.8%). The sensitivity to pollen grains displays preponderance (57.9%) to males. CONCLUSIONS: For the first time in Thessaloniki, Greece, 15-year allergenic pollen records were conducted. Clinical observations confirm that the pollen mainly implicated in respiratory allergy symptoms are grasses, olive and wall pellitory. PMID- 14763932 TI - Influence of a multidisciplinary paediatric allergy clinic on parental knowledge and rate of subsequent allergic reactions. AB - BACKGROUND: Studies have demonstrated that families of children with food allergy have significant deficiencies in their knowledge of how to avoid allergen exposure and how to manage allergic reactions. This study aims to assess the impact of a multidisciplinary paediatric allergy clinic consultation on parental knowledge of food allergy and to determine the rate of subsequent allergic reactions. METHODS: Sixty-two subjects (<17 years) referred with food allergy were prospectively enrolled. Parental knowledge was assessed by questionnaire and EpiPen trainer. Families saw a paediatric allergist, clinical nurse specialist and dietician. Knowledge was reassessed after 3 months and rate of allergic reactions after 1 year. RESULTS: After one visit to the paediatric allergy clinic, there was a significant improvement in parental knowledge of allergen avoidance (26.9%, P < 0.001), managing allergic reactions (185.4%, P < 0.0001) and EpiPen usage (83.3%, P < 0.001). Additionally, there was a significant reduction in allergic reactions (P < 0.001). Children with egg, milk or multiple food allergies were more likely to suffer subsequent reactions. CONCLUSIONS: A single visit to a multidisciplinary allergy clinic considerably improves families' abilities to manage allergic reactions to foods with an accompanying reduction in allergic reactions. Young children with egg, milk or multiple food allergies were at greatest risk of further reactions. PMID- 14763933 TI - Monosodium benzoate hypersensitivity in subjects with persistent rhinitis. AB - BACKGROUND: Very few data are available from the literature on whether nonatopic subjects affected by persistent rhinitis may show the appearance of objective symptoms of rhinitis after the ingestion of food additives such as tartrazine (E102), erythrosine (E127), monosodium benzoate (E211), p-hydroxybenzoate (E218), sodium metabisulphite (E223), and monosodium glutamate (E620). It is still unclear whether the ingestion of food additive may cause, as well, a consensual reduction of nasal peak inspiratory flow (NPIFR). Therefore, we used a double blind placebo-controlled (DBPC) study to evaluate this hypothesis. PATIENTS AND METHODS: Two hundred and twenty-six consecutive patients (76 males and 150 females) aged 12-60 years (mean age 40.2 +/- 16.3 years). After 1 month of an additive-free diet regimen, an open challenge was carried out (food additive-rich diet for 2 weeks). After this period, challenges were administered in a DBPC manner using the above-mentioned substances under investigation. RESULTS: Twenty of 226 subjects (8.8%) reported an improvement of the symptoms of rhinitis after additive-free diet. More precisely, six of 226 (2.6%) were symptom-free and 14 of 226 (6.2%) showed an improvement in their symptoms after an additive-free diet. As far as the results for DBPC are concerned, 20 challenges with monosodium benzoate induced both objective (i.e. sneezing and rhinorrhoea) and subjective symptoms (nasal blockage and nasal itching) of rhinitis with reduction of NPIFR >/=20%, 45 challenges induced subjective symptoms of rhinitis (i.e. nasal blockage and nasal itching), without reduction of NPIFR >/=20% of the basal value, two with tartrazine, seven with erythrosine, 19 with monosodium benzoate, three with p-hydroxybenzoate, six with sodium metabisulphite, and eight with monosodium glutamate, respectively. CONCLUSIONS: The observation that nonatopic persistent rhinitis may be caused by the frequent, probably daily, ingestion of small doses of a nontolerated substance is intriguing and suggests that at least some patients with 'chronic vasomotor rhinitis' may be intolerant to a particular food additive. Therefore, food additives can be considered triggers or aggravating factors, rather than aetiological factors. PMID- 14763934 TI - IgE antibody to Aspergillus fumigatus recombinant allergens in cystic fibrosis patients with allergic bronchopulmonary aspergillosis. AB - BACKGROUND: Allergic bronchopulmonary aspergillosis (ABPA) in cystic fibrosis (CF) is characterized by a heightened Th2 CD4+ T-cell response to Aspergillus fumigatus (Af) allergens and a hyper-immunoglobulin E (IgE) state compared with cystic fibrosis patients without ABPA. The IgE serologic differentiation of ABPA from atopic CF patients can be difficult. We propose as the reactivity with purified antigens varies qualitatively and quantitatively and that the antibody response is more specific than with crude Af antigen extract, the IgE responses to purified recombinant Af allergens may differentiate ABPA from atopic CF patients. METHODS: Serum IgE reactivity to seven recombinant purified allergens and to a crude extract of Af was measured in 15 ABPA, in 23 Af skin test positive (ST+), and in 19 Af skin test negative (ST-) CF patients. Four of the ABPA CF patients were studied before and after developing ABPA. Nine ABPA patients were studied during flares and remissions of ABPA. RESULTS: Allergic bronchopulmonary aspergillosis patients had significantly increased IgE reactivity to Asp f2, f3, f4, f6, and f16 compared with the Af ST+ and ST- non-ABPA CF patients. In the ABPA patients studied before and after developing ABPA, IgE reactivity also increased to Asp f2, f3, f4, and f6, and to the crude extract. In ABPA CF patients, IgE reactivity to Asp f1, f2, f3, and f6 significantly increased during periods of ABPA flares compared with periods of remission. Analysis of the receiver operating curve demonstrated that IgE reactivity to Asp f3 and f4 gave the best sensitivity and specificity and were better than IgE reactivity to a crude extract of Aspergillus. Furthermore, in ABPA patients studied during periods of remission the IgE reactivity to Asp f3 and f4 remained significantly elevated compared with Af ST+ non-ABPA patients. The IgE responses when considered either to be positive or negative to Asp f3 and f4 significantly differentiated ABPA from Af ST+ and ST- non-ABPA CF patients. In contrast, IgE reactivity was considered positive to the crude extract in 89% of ABPA, 61% of Af ST+, and 0% of Af ST- non-ABPA CF patients. CONCLUSIONS: Immunoglobulin E reactivity to a panel of purified Af allergens, especially to Asp f3 and f4, differentiates ABPA from atopic Af ST+ non-ABPA CF patients. Serial determinations of IgE reactivity to individual purified Aspergillus antigens, especially Asp f3, demonstrates that increases in IgE reactivity may provide improved distinction between stages of flares and remission compared with changes in IgE reactivity to a crude Aspergillus extract. PMID- 14763935 TI - Mucosal T-cell phenotypes in persistent atopic and nonatopic rhinitis show an association with mast cells. AB - BACKGROUND: Allergic rhinitis is characterized by selective expansion of T cell subsets with a CD4+ phenotype. Recently, we identified a subpopulation of nonallergic rhinitis subjects with increased epithelial mast cell and eosinophil populations, suggestive of local mucosal allergy. Previously, T cell subsets have not been characterized in this subselection of nonallergic subjects and furthermore, their relationship to mast cell and basophil effector cells remain unidentified. OBJECTIVE: To determine if a subpopulation of nonallergic subjects with idiopathic rhinitis (IR) have localized allergy confined to their nasal mucosa by comparing the T cell subsets and major histocompatibility complex (MHC) II expressing cells to persistent allergic rhinitis (PAR). Furthermore, the relationship between T cell subsets and mast cells/basophils was investigated. METHODS: None of the symptomatic patients in this study were clinically allergen challenged. Nasal turbinate mucosa was removed from patients with PAR, IR and normal controls. Morphometry was performed on immunostained sections for T cell subset populations including CD3+, CD4+, CD8+, CD25+, CD45RA+, CD45RO+, human leucocyte antigen (HLA)-DRalpha (MHC class II), mast cell tryptase and for basophils. RESULTS: Subjects with persistent allergic rhinitis differed to normal controls in showing significantly increased numbers of total (CD3+), activated (CD25+) and allergen-naive (CD45RA+) T lymphocytes in their nasal mucosa (P < 0.025). The naive CD45RA+ memory T cells correlated to mucosal mast cells in PAR (P = 0.03). IR patients differ to allergic subjects in showing significantly reduced numbers of epithelial HLA-DRalpha+ cells (P = 0.007), but increased numbers of CD8+ lymphocytes (P = 0.02). The CD8+ T cells correlated with mucosal mast cell numbers (P = 0.02). In both rhinitis groups, basophils were present in very low numbers obviating the need for statistical analysis. CONCLUSION: PAR is characterized by increased numbers of CD3+, CD25+ and CD45RA+ T lymphocytes compared with normal mucosa. Allergic and nonallergic rhinitis groups can be separated by significant differences in the number of epithelial antigen presenting cells (APCs) (HLA-DRalpha+) and sub-epithelial activated (CD25+) T cells. Moreover, IR patients do not significantly differ to their allergic counterparts with respect to total (CD3+) and naive (CD45RA+) T cell numbers, or numbers of epithelial activated (CD25+) lymphocytes. IR subjects show significantly increased numbers of CD8+ lymphocytes compared with control mucosa and although our findings suggest that the initiating inflammatory events may differ, both rhinitis groups show a similarity in pathology involving mucosal mast cells with an association to infiltrating T cells. PMID- 14763936 TI - Molecular analysis of sequence variants in the Fcepsilon receptor I beta gene and IL-4 gene promoter in Italian atopic families. AB - BACKGROUND: The genetic variants in the Fcepsilon receptor I beta gene (Glu237Gly) and the T allele of the (C590T) polymorphism of interleukin (IL)-4 gene promoter were reported to be associated with atopy. But the data of the studies in different populations are contrasting with one another. METHODS: A group of 25 Italian nuclear families were studied. In each family at least two allergic subjects were present. The allergic children were 65 and the allergic relatives were 35. One hundred and three nonallergic unrelated controls included outpatiens with no history of atopy. The (C590T) promoter polymorphism of the IL 4 and the genetic variant Glu237Gly of Fcepsilon RI beta genes were analysed by the polymerase chain reaction-restriction fragment length polymorphism method. RESULTS: A significant difference was observed in the genotype frequency at codon 237 of the Fcepsilon RI beta gene between allergic children and nonatopic control (P < 0.01) and in the allergic relatives (P < 0.001). In the children, the Glu237Gly polymorphism was also associated with elevated circulating levels of immunoglobulin E. The -590C/T allele of IL-4 promoter gene showed no association with atopy. CONCLUSIONS: In our study, the Glu237Gly polymorphism of the Fcepsilon RI beta gene was associated with atopy. Our results have not pointed out an association between the (C590T) promoter polymorphism of the IL-4 gene and atopy. These data suggest the potential role of the Fc RI beta gene in the development of the allergy. PMID- 14763937 TI - Skin test evaluation in nonimmediate allergic reactions to penicillins. AB - BACKGROUND: Nonimmediate reactions (NIR) to aminopenicillins (AP) are frequent. Although patch testing (PT) and intradermal testing (IT) are used for diagnosis, comparative results have never been adequately performed. We compared PT and IT in subjects with NIR to AP. METHODS: Twenty-one subjects with NIR to AP and positive IT were re-evaluated. Skin tests were performed with amoxicillin (AX) and ampicillin (AM) at different concentrations in petrolatum, 50, 25, and 5% w/w, for PT and in saline, 20, 2, and 0.2 mg/ml for both PT and IT. Skin biopsies from the site of the positive response were studied with haematoxylin-eosin and immunohistochemistry. RESULTS: In the re-evaluation, one case was IT and PT negative and was excluded; 20 were IT positive and 18 PT positive for both AX and AM. Decreasing concentrations of AP induced a reduction in positivity in both methods when diluted in saline, but not when mixed in petrolatum (PT only). With both PT and IT, immunohistochemical studies showed a perivascular mononuclear infiltrate with CD4 and CD8 memory cells expressing perforin and granzyme B. CONCLUSIONS: Both tests appear valuable for the diagnosis of NIR to AP. However, IT diagnosed more patients than PT. The vehicle (saline or petrolatum) had no influence on the response, although in the former the concentration was critical. The immunohistochemical analysis showed skin infiltrates compatible with a T-cell drug reaction. PMID- 14763938 TI - Cross-reactivity between obeche wood (Triplochiton scleroxylon) and natural rubber latex. AB - BACKGROUND: Several studies have shown that allergy to natural rubber latex is associated with cross reactivity to other plants. We have investigated a possible cross reactivity with a tree belonging to the Sterculiaceae family, Tryplochiton scleroxylon, in a patient allergic to obeche wood, that began to suffer symptoms of allergy in contact with latex. METHODS: Determinations of specific IgE by CAP to obeche, natural rubber latex (NRL) and recombinant allergens of latex were done. Immunoblotting was performed to study allergens detected by serum of our patient. CAP-inhibition methods were used to study cross-reactive between NRL and obeche wood. RESULTS: In obeche extract, 4 bands were found of an apparent molecular weight of 18, 38, 75 and 199 kDa. In NRL extract the allergens have a molecular weight around 18, 21, 32, 38, 60, 70 and 199 kDa. IgE to latex was almost completely inhibited (96%) by the obeche extract in CAP-inhibition studies. Using obeche extract as inhibitor, IgE binding to native hevein (nHevb 6.02) was reduced by 25%. CONCLUSIONS: The results obtained in CAP-inhibition demonstrate the existence of cross-reactivity between proteins in NRL and obeche wood. In our patient, a band of apparent molecular weight of 38 kDa could be the most important allergen. PMID- 14763939 TI - IgE antibody in the serum--the main problem is cross-reactivity. PMID- 14763941 TI - House dust mite allergens in Turkish homes. PMID- 14763943 TI - A human dermatitis caused by Thaumetopoea pityocampa (Denis and Schiffermuller, 1775) (order: Lepidoptera) caterpillars in Istanbul, Turkey. PMID- 14763942 TI - Lavender in pillows. No effect on Der p 1 accumulation. PMID- 14763944 TI - Levothyroxine-induced hyperthyroidism. PMID- 14763945 TI - 'Daily pattern' of an asthmatic reaction due to isocyanates. PMID- 14763946 TI - Contact dermatitis due to Edding 3000. PMID- 14763947 TI - Allergic bronchopulmonary aspergillosis without clinical asthma caused by Aspergillus niger. PMID- 14763948 TI - Fondaparinux as a novel therapeutic alternative in a patient with heparin allergy. PMID- 14763949 TI - A case of IgE-mediated hypersensitivity to cefepime. PMID- 14763950 TI - Anaphylaxis to dexrazoxane (ICRF-187) following three previous uncomplicated infusions. PMID- 14763951 TI - Valproic acid blood genomic expression patterns in children with epilepsy - a pilot study. AB - OBJECTIVE: Valproic acid (VPA) is a commonly used anticonvulsant with multiple systemic effects. The purpose of this pilot study is to examine the blood genomic expression pattern associated with VPA therapy in general and secondly VPA efficacy in children with epilepsy. MATERIALS AND METHODS: Using oligonucleotide microarrays, gene expression in whole blood was assessed in pediatric epilepsy patients following treatment with VPA compared with children with epilepsy prior to initiation of anticonvulsant therapy (drug free patients). RESULTS: The expression of 461 genes was altered in VPA patients (n = 11) compared with drug free patients (n = 7), among which a significant number of serine threonine kinases were down-regulated. Expression patterns in children seizure free on VPA therapy (n = 8) demonstrated 434 up-regulated genes, many in mitochondria, compared with VPA children with continuing seizures (n = 3) and drug free seizure patients (n = 7). CONCLUSION: VPA therapy is associated with two significant and unique blood gene expression patterns: chronic VPA monotherapy in general and a separate blood genomic profile correlated with seizure freedom. These expression patterns provide new insight into previously undetected mechanisms of VPA anticonvulsant activity. PMID- 14763952 TI - Herpesviral DNA in brain tissue from patients with temporal lobe epilepsy. AB - OBJECTIVES: Presence of DNA from six herpesviruses were examined in brain tissue from patients operated for temporal lobe epilepsy. MATERIAL AND METHODS: A total of 19 Canadian patients (I) with a median age of 22 years, 17 Swedish patients (II) with a median age of 14 years and a reference group comprising 12 individuals were studied. Presence of herpesviral DNA was detected by nested polymerase chain reaction. RESULTS: Of three children with Rasmussen's encephalitis, Cytomegalovirus (CMV) DNA was found in two, and human herpesvirus type 6 DNA in two. In six children with ganglioglioma, Epstein-Barr virus (EBV) was detected in four. CMV DNA was found significantly more in group I compared with II, while the reverse occurred with EBV DNA. Malformations of cortical development were found significantly more in group II compared with I. CONCLUSION: Detection of DNA from some herpesviruses in epileptic brain tissue may possibly be associated with distinct clinical conditions, but factors such as age and malformations of cortical development should also be considered. PMID- 14763953 TI - Diagnostic delays and mis-management in cluster headache. AB - OBJECTIVES: Cluster headache is a stereotyped form of primary headache that while common in terms of neurologic illnesses is much less common as a cause of disabling headache than migraine. MATERIALS AND METHODS: We directly interviewed 230 patients with cluster headache. National support groups contributed 76% and 24% came from the National Hospital for Neurology and Neurosurgery Headache Clinic. RESULTS: Seventy-two percent were men and 28% women, giving a male to female (M:F) ratio of 2.5:1. Episodic cluster headache (ECH) was recorded in 79% while 21% had chronic cluster headache (CCH). The mean time to diagnosis has dropped from 22 years in the 1960s to 2.6 years in the 1990s, although the mean number of GPs seen before a diagnosis was made remains at three. CONCLUSIONS: While there has been improvement in the time to diagnosis for cluster headache, a number of physicians will be consulted, and better education is likely to reduce the overall patient suffering. PMID- 14763954 TI - Is obstructive sleep apnea syndrome associated with headache? AB - OBJECTIVE: Obstructive sleep apnea syndrome (OSA) is a common disorder in the general population. Although the mechanisms remain obscure, an association with headache has been reported. We aimed to assess the frequency of OSA in a population of headache patients based on a stratified sampling technique using questionnaire and polysomnography (PSG). METHODS: The survey was provided to new outpatients examined by a neurologist for headache over a 2-year period of time. The patients completed a headache diary during 30 days and those at risk of OSA went through a PSG examination. RESULTS: Of 903 headache patients, 75 reported heavy snoring and episodes of interrupted nocturnal breathing (8%). Among 43 patients examined with PSG, 14 (1.5% of the total study population) had an apnea/hypopnea index of 5 or higher. Eleven of the patients reported morning headache. CONCLUSION: The frequency of OSA in a patients referred to specialist for headache problems is not higher than what is reported for the general population. The relatively low rate of OSA in this selected group of patients with headache referred to neurology for second opinion does not support the notion that OSA brings about headache. PMID- 14763955 TI - Spinocerebellar ataxia type 17 in the Yugoslav population. AB - OBJECTIVES: (1) Analysis of Spinocerebellar ataxia type 17 (SCA17) locus in a group of ataxic patients excluded on other known SCAs; (2) assessment of frequency distributions of SCA17 alleles in the Yugoslav population. MATERIAL AND METHODS: Study includes 115 non-related Yugoslav patients belonging to autosomal dominant cerebellar ataxias or to sporadic idiopathic adult-onset ataxia and 115 controls. Analysis of SCA17 locus was performed using polymerase chain reaction. RESULTS: None of the analyzed patients show the presence of mutation in SCA17 locus. In the group of patients 12 different alleles in the range of 30-42 repeats were observed, while in healthy population eight alleles in the range of 30-40 repeats were detected. CONCLUSION: (1) None of 115 non-related Yugoslav ataxic patients belong to any known SCAs nor to DRPLA gene; (2) the distribution of SCA17 alleles in the Yugoslav population is consistent with the distribution in other populations and (3) the paucity of alleles with more than 39 repeats could suggest that SCA17 is very rare in the Yugoslav population. PMID- 14763956 TI - Thalidomide neuropathy: clinical, electrophysiological and neuroradiological features. AB - OBJECTIVE: Thalidomide is a promising therapy for multiple myeloma. Sensory neuropathy is a side effect of thalidomide and resulted to be partially reversible in 50% of cases, suggesting a sensory ganglionopathy. Spinal cord magnetic resonance imaging (MRI) was found to be useful in the diagnosis of sensory ganglionopathies and we use it to determine if thalidomide neuropathy has features of a ganglionopathy. MATERIAL AND METHODS: Six patients with multiple myeloma developed thalidomide-induced polyneuropathy. Nerve conduction studies, somatosensory-evoked potentials (SEPs) and cervical and dorsal spinal cord MRI were obtained in all. RESULTS: All patients had a sensory neuropathy, with clinical or electrophysiological abnormalities involving all four limbs. Spinal cord MRI showed high signal intensity in the posterior columns in only one patient, with abnormal central conduction time at SEPs. CONCLUSION: Our results suggest that thalidomide can induce either an axonal length-dependent neuropathy or, less frequently, a ganglionopathy. PMID- 14763957 TI - Sensory perception in complete spinal cord injury. AB - OBJECTIVES: To describe sensations evoked by painful or repetitive stimulation below injury level in patients with a clinically complete (American Spinal Injury Association, ASIA Grade A) spinal cord injury (SCI). MATERIAL AND METHODS: Twenty four patients (11 with central neuropathic pain and 13 without pain) with a traumatic SCI above the tenth thoracic vertebra were examined using quantitative sensory testing, MR imaging, and somatosensory evoked potentials (SEP). RESULTS: Painful (pressure, pinch, heat or cold) or repetitive (pinprick) stimuli elicited vague localized sensations in 12 patients (50%). Pain, spasticity, and spasms were equally seen in SCI patients with or without localized sensations. SEP and MRI did not differentiate between these two groups. CONCLUSION: The present study suggests retained sensory communication across the injury in complete SCI, i.e. 'sensory discomplete' SCI. PMID- 14763958 TI - Blood pressure, nimodipine, and outcome of ischemic stroke. AB - OBJECTIVES: The reduction of blood pressure (BP) caused by nimodipine has been proposed as an explanation for the poor results in ischemic stroke trials. We evaluated further the relationships between BP, nimodipine, and outcome of ischemic stroke, and also searched for other possible explaining mechanisms. PATIENTS AND METHODS: All 350 participants of an earlier placebo controlled trial on oral nimodipine were included in this study. Among other variables, the admission BP, and the change of BP during the first day were noted. The 3 week and 3 month functional outcome was assessed with a modified Rankin grading. RESULTS: The severity of stroke was the utmost important predictor of outcome. Visible cerebral infarction on computed tomography (CT) was associated with severe stroke and an early commencement (within 24 h of stroke onset) of nimodipine treatment. In the nimodipine arm, high initial systolic and diastolic BP measured < or =24 h of stroke onset were independent predictors of good functional outcome (Rankin grades 1 and 2), whereas BP change was not. The survivors in the nimodipine arm with mild to moderately severe stroke had higher initial BP than the deceased ones, in severe strokes the situation was the opposite. CONCLUSIONS: Stroke severity, visible cerebral infarcts on CT, and early commencement of nimodipine treatment were associated. Overall, high initial systolic and diastolic BP predicted a good functional outcome in patients on nimodipine. In severe strokes, the combination of nimodipine and high initial BP was associated with increased risk of death. PMID- 14763959 TI - Apolipoprotein E polymorphism and neuropsychological outcome following subarachnoid haemorrhage. AB - OBJECTIVES: To investigate the association between APOE genotype and cognitive and emotional outcome following spontaneous subarachnoid haemorrhage (SAH). MATERIALS AND METHODS: Neuropsychological assessments were conducted with 70 SAH survivors derived from a consecutive series of neurosurgical admissions. Outcomes, including cognitive tests, health questionnaires and Glasgow Outcome Scale at a mean of 16 months after SAH, were compared with presence or absence of the epsilon4 allele. RESULTS: There was no evidence that SAH survivors possessing the epsilon4 allele had poorer outcome. The only suggestion of an association between the epsilon4 allele and outcome was in a subgroup of patients with a Fisher grade 4 haemorrhage, although this trend did not reach statistical significance. CONCLUSIONS: Overall, possession of the APOE epsilon4 allele is not significantly associated with neuropsychological outcome following SAH. However, there may be an effect amongst those with a Fisher grade 4 haemorrhage. PMID- 14763960 TI - M-wave analysis and passive tilt in patients with different degrees of carotid artery disease. AB - BACKGROUND AND PURPOSE: Carotid artery disease (CAD) is able to critically impair cerebral autoregulation which increases the risk for stroke. As therapeutic strategy largely depends on the degree of CAD, we investigated whether this gradation is also related to significant changes in autoregulatory capacity. We applied cross-spectral analysis (CSA) of spontaneous Mayer-wave (M-wave) oscillations and passive tilting (PT) to test cerebral autoregulation. METHODS: Cerebral autoregulation was tested in 102 patients with carotid stenosis (> or =70%) or occlusion and 14 controls by comparison of continuous transcranial Doppler sonography of the middle cerebral artery and beat-to-beat arterial blood pressure (ABP) during PT to 80 degrees head-up position as well as by CSA of M waves (3-9 cpm). RESULTS: The orthostatic decrease of cerebral blood flow velocity (CBFV) was not correlated with the degree of CAD and showed a lower sensitivity and specificity than phase angle shifts between M-waves in ABP and CBFV (sensitivity: 75-80%, specificity: 86%). Phase angles were gradually lowered in carotid stenoses > 70%, but apparently, they were only moderately correlated with the degree of CAD (r = -0.35, P < 0.01). An additional influencing factor seemed to be the sufficiency of collateralization. CONCLUSIONS: The results show that CSA of M-waves is more appropriate for testing autoregulation than PT. CSA suggests that the capacity to autoregulate depends to a certain extent on the degree of CAD but is also influenced by the sufficiency of collateral pathways and pre-existing strokes. PMID- 14763961 TI - Response of thymectomy: clinical and pathological characteristics among seronegative and seropositive myasthenia gravis patients. AB - OBJECTIVE: To identify the response to thymectomy in patients with seronegative and seropositive myasthenia gravis (SPMG). We analyzed the associated diseases, thymus histology, and the severity of symptoms between the two groups. MATERIAL AND METHODS - DESIGN: Descriptive, comparative. STUDY UNITS: Fourteen patients with seronegative myasthenia gravis (SNMG) and 57 patients with SPMG who had a thymectomy between 1987 and 1997, with at least 3 years of follow-up. The patients were divided into four groups; (1) Remission, (2) Improvement, (3) No change and (4) Deterioration. RESULTS: Fourteen patients (20%) were seronegative and 57 were seropositive (80%). In the group of patients with SNMG, three patients were in remission (21%), five with improvement (36%), five with no change (36%) and one with worsening (7%). In the group of patients with SPMG, 12 were in remission (21%), 17 with improvement (30%), 25 with no change (44%) and three (5%) with worsening. The patients with SNMG were older, with less associated diseases and with a lower frequency of thymomas. CONCLUSIONS: The response to thymectomy was similar between the two groups. It has been suggested that seronegative patients have a better prognosis, but our results show no differences. PMID- 14763963 TI - Prevalence of Parkinson's disease in a Chinese population. AB - BACKGROUND: Ethnic differences in the prevalence of Parkinson's disease have been observed, but may be due to differences in screening instruments and diagnostic criteria. AIMS: To compare the prevalence of Parkinson's disease in Hong Kong Chinese and Australians in Sydney. METHODS: A door-to-door community survey in the Hong Kong Chinese population was carried out using a two-staged procedure (screening questionnaire followed by physical examination), which had been validated for use in the community in a survey among Australians, to determine the prevalence in Chinese and to compare with that in Australians. RESULTS: A total of 1080 households were contacted, the non-response rate was 27%. The prevalence among those aged 55 years in the community and above was 0.5%, and 0.186% for the overall population assuming no subjects were below 55 years. These figures are lower than the 3.6% for Australians, aged >55 years. CONCLUSION: There appears to be a difference in prevalence of Parkinson's disease between Hong Kong Chinese and Australians in Sydney. Such differences may be due to documented differences in prevalence of genetic polymorphisms associated with Parkinson's disease between Chinese and Australians, or to differences in environmental factors. PMID- 14763962 TI - Specific APO E genotypes in combination with the ACE D/D or MTHFR 677TT mutation yield an independent genetic risk of leukoaraiosis. AB - OBJECTIVE: Ischaemic demyelination of the white matter of the brain is a frequent clinical entity. In the neuroimaging terms, it is referred to as leukoaraiosis. We earlier found that the co-occurrence of the homozygous methylenetetrahydrofolate reductase (MTHFR) 677TT and angiotensin-converting enzyme D/D (ACE D/D) genotypes yielded a highly significant moderate risk of leukoaraiosis. On the assumption of further genetic interactions, we have now investigated whether the different apolipoprotein E (APO E) genotypes, in pairwise combinations with the MTHFR 677TT or ACE D/D mutation, could lead to an increased risk of leukoaraiosis. MATERIAL AND METHODS: We analysed the occurrence of the APO E genotypes in pairwise combinations with the MTHFR 677TT or ACE D/D mutation in 315 consecutive Caucasian patients with leukoaraiosis. A total of 646 neuroimaging-free subjects acted as a control group. RESULTS: The APO E 2/2 and 2/3 or APO E 4/4 and 4/3 genotypes in combination with the MTHFR 677TT or ACE D/D mutation exhibited independent genetic risks of leukoaraiosis. CONCLUSION: The interactions of certain unfavourable genetic mutations can contribute to the evolution of leukoaraiosis. PMID- 14763964 TI - Reflex writing seizures in two siblings with juvenile myoclonic epilepsy. AB - We report on two siblings who presented with juvenile myoclonic epilepsy, and in whom myoclonic jerks of the right arm and hand were also triggered by writing tasks. Both patients underwent intensive video-electroencephalography monitoring, with simultaneous neuropsychological tests. In both patients, reflex epileptic myoclonus was more easily triggered by writing that required a higher degree of concentration. Conversely, other cognitive tasks, such as reading, typing, thinking, or calculation never elicited any seizures or myoclonus. Valproate was effective in controlling both spontaneous and reflex epileptic seizures. The results of this study further support the notion that 'praxis-induced' reflex epilepsy precipitated by specific stimuli occurs in the context of idiopathic generalized epilepsy. Our results also illustrate that writing tasks are more effective in eliciting seizures when they require higher levels of concentration and mental elaboration. PMID- 14763965 TI - Kynurenic acid in ALS patients. PMID- 14763968 TI - Let there be blight: functional analysis of virulence in Phytophthora infestans. PMID- 14763969 TI - Cellular engineering in a minimal microbe: structure and assembly of the terminal organelle of Mycoplasma pneumoniae. AB - Mycoplasma pneumoniae is a minimal microbe with respect to cell envelope composition, biosynthetic and regulatory capabilities and genome size, yet it possesses a remarkably complex, multifunctional terminal organelle. This membrane bound extension of the mycoplasma cell is defined by the presence of an electron dense core that appears as paired, parallel bars oriented longitudinally and enlarging at the distal end to form a terminal button. Most non-cytadhering mutants of M. pneumoniae isolated to date exhibit defects in the architecture of the terminal organelle. Detailed characterization of those mutants has revealed the identities of many component proteins of the terminal organelle as well as the likely order in which some of those components are required. Additional questions regarding the composition of the electron-dense core, the means by which the terminal organelle is duplicated during cell division and the manner in which this process is regulated remain to be answered. Thus, it seems that there is much to be learned about cellular engineering and spatial regulation in these 'simple' cell wall-less bacteria. PMID- 14763970 TI - A Galpha subunit controls zoospore motility and virulence in the potato late blight pathogen Phytophthora infestans. AB - The heterotrimeric G-protein pathway is a ubiquitous eukaryotic signalling module that is known to regulate growth and differentiation in many plant pathogens. We previously identified Pigpa1, a gene encoding a G-protein alpha subunit from the potato late blight pathogen Phytophthora infestans. P. infestans belongs to the class oomycetes, a group of organisms in which signal transduction processes have not yet been studied at the molecular level. To elucidate the function of Pigpa1, PiGPA1-deficient mutants were obtained by homology-dependent gene silencing. The Pigpa1-silenced mutants produced zoospores that turned six to eight times more frequently, causing them to swim only short distances compared with wild type. Attraction to the surface, a phenomenon known as negative geotaxis, was impaired in the mutant zoospores, as well as autoaggregation and chemotaxis towards glutamic and aspartic acid. Zoospore production was reduced by 20-45% in different Pigpa1-silenced mutants. Transformants expressing constitutively active forms of PiGPA1, containing amino acid substitutions (R177H and Q203L), showed no obvious phenotypic differences from the wild-type strain. Infection efficiencies on potato leaves ranged from 3% to 14% in the Pigpa1-silenced mutants, compared with 77% in wild type, showing that virulence is severely impaired. The results prove that PiGPA1 is crucial for zoospore motility and for pathogenicity in an important oomycete plant pathogen. PMID- 14763971 TI - Activating mutations of Tn3 resolvase marking interfaces important in recombination catalysis and its regulation. AB - Catalysis of DNA recombination by Tn3 resolvase is conditional on prior formation of a synapse, comprising 12 resolvase subunits and two recombination sites (res). Each res binds a resolvase dimer at site I, where strand exchange takes place, and additional dimers at two adjacent 'accessory' binding sites II and III. 'Hyperactive' resolvase mutants, that catalyse strand exchange at site I without accessory sites, were selected in E. coli. Some single mutants can resolve a res x site I plasmid (that is, with one res and one site I), but two or more activating mutations are necessary for efficient resolution of a site I x site I plasmid. Site I x site I resolution by hyperactive mutants can be further stimulated by mutations at the crystallographic 2-3' interface that abolish activity of wild-type resolvase. Activating mutations may allow regulatory mechanisms of the wild-type system to be bypassed, by stabilizing or destabilizing interfaces within and between subunits in the synapse. The positions and characteristics of the mutations support a mechanism for strand exchange by serine recombinases in which the DNA is on the outside of a recombinase tetramer, and the tertiary/quaternary structure of the tetramer is reconfigured. PMID- 14763972 TI - The high-resolution functional map of bacteriophage SPP1 portal protein. AB - An essential component in the assembly of nucleocapsids of tailed bacteriophages and of herpes viruses is the portal protein that is located at the unique vertex of the icosahedral capsid through which DNA movements occur. A library of mutations in the bacteriophage SPP1 portal protein (gp6) was generated by random mutagenesis of gene 6. Screening of the library allowed identification of 67 single amino acid substitutions that impair portal protein function. Most of the mutations cluster within stretches of a few amino acids in the gp6 carboxyl terminus. The mutations were divided into five classes according to the step of virus assembly that they impair: (1) production of stable gp6; (2) interaction of gp6 with the minor capsid protein gp7; (3) incorporation of gp6 in the procapsid structure; (4) DNA packaging; and (5) sizing of the packaged DNA molecule. Most of the mutations fell in classes 3 and 4. This is the first high-resolution functional map of a portal protein, in which its function at different steps of viral assembly can be directly correlated with specific regions of its sequence. The work provides a framework for the understanding of central processes in the assembly of viruses that use specialized portals to govern entry and exit of DNA from the viral capsid. PMID- 14763973 TI - Structural studies of Streptococcus pneumoniae EPSP synthase in unliganded state, tetrahedral intermediate-bound state and S3P-GLP-bound state. AB - The shikimate pathway synthesizes aromatic amino acids and other essential metabolites that are necessary for bacteria, plants and fungi to survive. This pathway is not present in vertebrates and therefore represents an attractive target for antibacterial agents. We have successfully crystallized and solved the structure of unliganded, inhibitor-liganded and tetrahedral intermediate (TI) liganded forms of Streptococcus pneumoniae EPSP synthase. The overall topology of the S. pneumoniae EPSP synthase is similar to that of the Escherichia coli EPSP synthase. In addition, the majority of residues responsible for ligand binding were conserved between the two proteins. TI-liganded structure provides absolute configuration of the C-2 atom from the F-PEP moiety of the enzyme-bound intermediate and also defines key residues responsible for the enzyme reaction. Comparison of the unliganded state and substrate-bound state of the enzyme provides insights into the structural mechanisms involved in dynamic events of ligand binding, domain movement and closure. This structural study of the pathogenic bacteria S. pneumoniae EPSP synthase with inhibitor and TI will provide invaluable information for the design of new-generation antibiotics. PMID- 14763974 TI - The Pseudomonas aeruginosa RpoS regulon and its relationship to quorum sensing. AB - In Escherichia coli and some other gamma-Proteobacteria, the alternative sigma factor RpoS functions as a regulator of the general stress response. The role of RpoS in Pseudomonas aeruginosa is not clear. Although P. aeruginosa RpoS contributes to the resistance to several environmental stresses, its role appears to be less pivotal than in E. coli. In P. aeruginosa, RpoS also regulates the production of several virulence factors and influences the expression of individual genes that are controlled by quorum sensing. Some quorum-controlled genes are induced by RpoS, whereas others are repressed. To gain insights about RpoS function in P. aeruginosa and to understand better the regulation of quorum controlled genes, we used transcript profiling to define an RpoS regulon. We identified 772 genes regulated by RpoS in stationary but not in logarithmic growth phase (504 were induced and 268 were repressed), and we identified putative RpoS promoter sequence elements with similarity to the E. coli RpoS consensus in several of these genes. Many genes in the regulon, for example a set of chemotaxis genes, have assigned functions that are distinct from those in E. coli and are not obviously related to a stress response. Furthermore, RpoS affects the expression of more than 40% of all quorum-controlled genes identified in our previous transcriptome analysis. This highlights the significance of RpoS as a global factor that controls quorum-sensing gene expression at the onset of stationary phase. The transcription profiling results have allowed us to build a model that accommodates previous seemingly conflicting reports. PMID- 14763975 TI - Dynamics and processivity of 40S ribosome scanning on mRNA in yeast. AB - The eukaryotic 40S ribosomal subunit locates the translation initiation codon on an mRNA via the so-called scanning process that follows 40S binding to the capped 5' end. This key step in translation is required for the expression of almost all eukaryotic genes, yet the mechanism and dynamics of scanning are unknown. We have performed quantitative studies in vivo and in vitro of the movement of yeast 40S ribosomes along 5' untranslated regions (UTRs) of different lengths. 40S subunits perform cap-dependent scanning with high processivity for more than 1700 nucleotides in cells of Saccharomyces cerevisiae. Moreover, the observed rates of expression indicate that scanning is performed by an untethered 40S subunit that has been released from the 5' cap complex. Unexpectedly, the capability to maintain scanning competence on a long 5' UTR is more dependent on the Ded1/Dbp1 type of helicase than on eIF4A or eIF4B. In a yeast cell-free extract, scanning shows reduced processivity, with an estimated net 5'-->3' rate of approximately 10 nucleotides per second at 26 degrees C. We have developed a biased bidirectional walking model of ribosomal scanning that provides a framework for understanding the above observations as well as other known quantitative and qualitative features of this process. PMID- 14763976 TI - The Mycobacterium tuberculosis ino1 gene is essential for growth and virulence. AB - Inositol is utilized by Mycobacterium tuberculosis in the production of its major thiol and of essential cell wall lipoglycans. We have constructed a mutant lacking the gene encoding inositol-1-phosphate synthase (ino1), which catalyses the first committed step in inositol synthesis. This mutant is only viable in the presence of extremely high levels of inositol. Mutant bacteria cultured in inositol-free medium for four weeks showed a reduction in levels of mycothiol, but phosphatidylinositol mannoside, lipomannan and lipoarabinomannan levels were not altered. The ino1 mutant was attenuated in resting macrophages and in SCID mice. We used site-directed mutagenesis to alter four putative active site residues; all four alterations resulted in a loss of activity, and we demonstrated that a D310N mutation caused loss of the active site Zn2+ ion and a conformational change in the NAD+ cofactor. PMID- 14763977 TI - The galactokinase of Hypocrea jecorina is essential for cellulase induction by lactose but dispensable for growth on d-galactose. AB - Lactose is the only soluble carbon source which can be used economically for the production of cellulases or heterologous proteins under cellulase expression signals by Hypocrea jecorina (=Trichoderma reesei). Towards an understanding of lactose metabolism and its role in cellulase formation, we have cloned and characterized the gal1 (galactokinase) gene of H. jecorina, which catalyses the first step in d-galactose catabolism. It exhibits a calculated Mr of 57 kDa, and shows moderate identity (about 40%) to its putative homologues of Saccharomyces cerevisiae and Kluyveromyces lactis. Gal1 is a member of the GHMP family, shows conservation of a Gly/Ser rich region involved in ATP binding and of amino acids (Arg 51, Glu 57, Asp 60, Asp 214, Tyr 270) responsible for galactose binding. A single transcript was formed constitutively during the rapid growth phase on all carbon sources investigated and accumulated to about twice this level during growth on d-galactose, l-arabinose and their corresponding polyols. Deletion of gal1 reduces growth on d-galactose but does only slightly affect growth on lactose. This is the result of the operation of a second pathway for d-galactose catabolism, which involves galactitol as an intermediate, and whose transient concentration is strongly enhanced in the delta-gal1 strain. In this pathway, galactitol is catabolised by the lad1-encoded l-arabinitol-4-dehydrogenase, because a gal1/lad1 double delta-mutant failed to grow on d-galactose. In the delta-gal1 strain, induction of the Leloir pathway gene gal7 (encoding galactose 1-phosphate uridylyltransferase) by d-galactose, but not by l-arabinose, is impaired. Induction of cellulase gene expression by lactose is also impaired in a gal1 deleted strain, whereas their induction by sophorose (the putative cellulose derived inducer) was shown to be normal, thus demonstrating that galactokinase is a key enzyme for cellulase induction during growth on lactose, and that induction by lactose and sophorose involves different mechanisms. PMID- 14763978 TI - Structure of the OmpA-like domain of RmpM from Neisseria meningitidis. AB - RmpM is a putative peptidoglycan binding protein from Neisseria meningitidis that has been shown to interact with integral outer membrane proteins such as porins and TonB-dependent transporters. Here we report the 1.9 A crystal structure of the C-terminal domain of RmpM. The 150-residue domain adopts a betaalphabetaalphabetabeta fold, as first identified in Bacillus subtilis chorismate mutase. The C-terminal RmpM domain is homologous to the periplasmic, C terminal domain of Escherichia coli OmpA; these domains are thought to be responsible for non-covalent interactions with peptidoglycan. From the structure of the OmpA-like domain of RmpM, we suggest a putative peptidoglycan binding site and identify residues that may be essential for binding. Both the crystal structure and solution experiments indicate that RmpM may exist as a dimer. This would promote more efficient peptidoglycan binding, by allowing RmpM to interact simultaneously with two glycan chains through its C-terminal, OmpA-like binding domain, while its (structurally uncharacterized) N-terminal domain could stabilize oligomers of porins and TonB-dependent transporters in the outer membrane. PMID- 14763979 TI - Divergent binding sites on intercellular adhesion molecule-1 (ICAM-1) for variant Plasmodium falciparum isolates. AB - Adhesion of human erythrocytes infected with the malaria parasite Plasmodium falciparum to host endothelium has been associated with severe forms of this disease. A number of endothelial receptors have been identified, and there is evidence that one of these, intercellular adhesion molecule-1 (ICAM-1), may play an important role in the pathology of cerebral malaria. Mutagenesis of domain 1 of ICAM-1, which is involved in parasite adhesion, shows that the binding sites for different parasite variants overlap to a large extent, but that there are subtle differences between them that correlate with their adhesive phenotypes. This suggests that the ability to bind to ICAM-1 has arisen from a common variant, but that subsequent changes have led to differences in binding avidity, which may affect pathogenesis. The definition of common binding determinants and the elucidation of links between ICAM-1 binding phenotype and disease will provide new leads in the design of therapeutic interventions. PMID- 14763980 TI - Identification of competence pheromone responsive genes in Streptococcus pneumoniae by use of DNA microarrays. AB - Natural genetic transformation in Streptococcus pneumoniae is controlled in part by a quorum-sensing system mediated by a peptide pheromone called competence stimulating peptide (CSP), which acts to coordinate transient activation of genes required for competence. To characterize the transcriptional response and regulatory events occurring when cells are exposed to competence pheromone, we constructed DNA microarrays and analysed the temporal expression profiles of 1817 among the 2129 unique predicted open reading frames present in the S. pneumoniae TIGR4 genome (84%). After CSP stimulation, responsive genes exhibited four temporally distinct expression profiles: early, late and delayed gene induction, and gene repression. At least eight early genes participate in competence regulation including comX, which encodes an alternative sigma factor. Late genes were dependent on ComX for CSP-induced expression, many playing important roles in transformation. Genes in the delayed class (third temporal wave) appear to be stress related. Genes repressed during the CSP response include ribosomal protein loci and other genes involved in protein synthesis. This study increased the number of identified CSP-responsive genes from approximately 40 to 188. Given the relatively large number of induced genes (6% of the genome), it was of interest to determine which genes provide functions essential to transformation. Many of the induced loci were subjected to gene disruption mutagenesis, allowing us to establish that among 124 CSP-inducible genes, 67 were individually dispensable for transformation, whereas 23 were required for transformation. PMID- 14763981 TI - Interconnection of competence, stress and CiaR regulons in Streptococcus pneumoniae: competence triggers stationary phase autolysis of ciaR mutant cells. AB - Of the 13 two-component signal transduction systems (TCS) identified in Streptococcus pneumoniae, two, ComDE and CiaRH, are known to affect competence for natural genetic transformation. ComD and ComE act together with the comC encoded competence-stimulating peptide (CSP) and with ComAB, the CSP-dedicated exporter, to co-ordinate activation of genes required for differentiation to competence. Several lines of evidence suggest that the CiaRH TCS and competence regulation are interconnected, including the observation that inactivation of the CiaR response regulator derepresses competence. However, the nature of the interconnection remains poorly understood. Interpretation of previous transcriptome analyses of ciaR mutants was complicated by competence derepression in the mutants. To circumvent this problem, we have used microarray analysis to investigate the transition from non-competence to competence in a comC-null wild type strain and its ciaR derivative after the addition of CSP. This study increased the number of known CSP-induced genes from approximately 47 to 105 and revealed approximately 42 genes with reduced expression in competent cells. Induction of the CiaR regulon, as well as the entire HrcA and part of the CtsR stress response regulons, was observed in wild-type competent cells. Enhanced induction of stress response genes was detected in ciaR competent cells. In line with these observations, CSP was demonstrated to trigger growth arrest and stationary phase autolysis in ciaR cells. Taken together, these data strongly suggest that differentiation to competence imposes a temporary stress on cells, and that the CiaRH TCS is required for the cells to exit normally from the competent state. PMID- 14763982 TI - MurAA, catalysing the first committed step in peptidoglycan biosynthesis, is a target of Clp-dependent proteolysis in Bacillus subtilis. AB - The carboxyvinyl transfer from phosphoenolpyruvate to UDP-N-acetylglucosamine is the first committed step in the pathway of peptidoglycan formation. This crucial reaction for bacterial cell growth is catalysed by the MurA enzymes. Gram negative bacteria carry one murA gene, whereas in a subgroup of Gram-positive bacteria two separate paralogues, MurAA and MurAB, exist. This study provides evidence that in the Gram-positive bacterium Bacillus subtilis, the MurAA protein is specifically degraded by the ClpCP protease. This Clp-dependent degradation is especially enhanced upon entry into stationary phase, thus ensuring an immediate growth arrest due to stalled murein biosynthesis. The MurAA protein can therefore be addressed as a target of Clp-dependent regulatory proteolysis such as the transcriptional regulators CtsR, ComK, Spx in B. subtilis, CtrA in Caulobacter crescentus or RpoS in Escherichia coli. Taking into account all other known regulatory targets of ATP-dependent proteases, MurAA of B. subtilis represents the first example of a metabolic enzyme which is a unique regulatory substrate of Clp-dependent proteolysis. Its function as a regulatory metabolic checkpoint resembles that of homoserine trans-succinylase (MetA) in E. coli which is similarly ATP-dependently degraded. PMID- 14763983 TI - Signal quenching, detoxification and mineralization of vir gene-inducing phenolics by the VirH2 protein of Agrobacterium tumefaciens. AB - Plant tumorigenesis by Agrobacterium tumefaciens requires approximately 20 Vir proteins, transcription of which is induced by a family of phenolic compounds released from plant wound sites. One Vir protein, VirH2, plays a role in the metabolism of at least one phenolic inducer inasmuch as it converts ferulic acid, a potent vir gene inducer, to the non-inducer caffeate by O-demethylation of a methoxyl group. Here, we tested VirH2-dependent O-demethylation of 16 other vir inducing phenolics, and detected this activity for each compound. However, O demethylation rates differed enormously, with the strongest vir gene inducers such as acetosyringone being demethylated extremely slowly. Compounds containing two methoxyl groups were demethylated at both positions. In general, phenolic inducers were more toxic than their demethylated counterparts. A virH2 mutant was more sensitive than the wild type to growth inhibition by virtually all phenolic inducers tested, indicating that VirH2 detoxifies these compounds. VirH2 also played a role in mineralization of some phenolics. It converted vanillate to protocatechuate, which was then mineralized via the beta-ketoadipate pathway. Vanillyl alcohol and vanillin were also mineralized after being oxidized to vanillate. All three compounds served as sole sources of carbon, whereas the remaining 13 compounds did not. PMID- 14763984 TI - Metabolic block at early stages of the glycolytic pathway activates the Rcs phosphorelay system via increased synthesis of dTDP-glucose in Escherichia coli. AB - A mutational block in the early stages of the glycolytic pathway facilitates the degradation of the ptsG mRNA encoding the major glucose transporter IICBGlc in Escherichia coli. The degradation is RNase E dependent and is correlated with the accumulation of either glucose-6-P or fructose-6-P (Kimata et al., 2001, EMBO J 20: 3587-3595; Morita et al., 2003, J Biol Chem 278: 15608-15614). In this paper, we investigate additional physiological effects resulting from the accumulation of glucose-6-P caused by a mutation in pgi encoding phosphoglucose isomerase, focusing on changes in gene expression. The addition of glucose to the pgi strain caused significant growth inhibition, in particular in the mlc background. Cell growth then gradually resumed as the level of IICBGlc decreased. We found that the transcription of the cps operon, encoding a series of proteins responsible for the synthesis of colanic acid, was markedly but transiently induced under this metabolic stress. Both genetic and biochemical studies revealed that the metabolic stress induces cps transcription by activating the RcsC/YojN/RcsB signal transduction system. Overexpression of glucose-6-P dehydrogenase eliminated both growth inhibition and cps induction by reducing the glucose-6-P level. Mutations in genes responsible for the synthesis of glucose-1-P and/or dTDP-glucose eliminated the activation of the Rcs system by the metabolic stress. Taken together, we conclude that an increased synthesis of dTDP-glucose activates the Rcs phosphorelay system, presumably by affecting the synthesis of oligosaccharides for enterobacterial common antigen and O-antigen. PMID- 14763985 TI - Overexpression of Upf1p compensates for mitochondrial splicing deficiency independently of its role in mRNA surveillance. AB - In yeast the UPF1, UPF2 and UPF3 genes encode three interacting factors involved in translation termination and nonsense-mediated mRNA decay (NMD). UPF1 plays a central role in both processes. In addition, UPF1 was originally isolated as a multicopy suppressor of mitochondrial splicing deficiency, and its deletion leads to an impairment in respiratory growth. Here, we provide evidence that inactivation of UPF2 or UPF3, like that of UPF1, leads to an impairment in respiratory competence, suggesting that their products, Upf1p, Upf2p and Upf3p, are equivalently involved in mitochondrial biogenesis. In addition, however, we show that only Upf1p acts as a multicopy suppressor of mitochondrial splicing deficiency, and its activity does not require either Upf2p or Upf3p. Mutations in the conserved cysteine- and histidine-rich regions and ATPase and helicase motifs of Upf1p separate the ability of Upf1p to complement the respiratory impairment of a Deltaupf1 strain from its ability to act as a multicopy suppressor of mitochondrial splicing deficiency, indicating that distinct pathways express these phenotypes. In addition, we show that, when overexpressed, Upf1p is not detected within mitochondria, suggesting that its role as multicopy suppressor of mitochondrial splicing deficiency is indirect. Furthermore, we provide evidence that cells overexpressing certain upf1 alleles accumulate a phosphorylated isoform of Upf1p. Altogether, these results indicate that overexpression of Upf1p compensates for mitochondrial splicing deficiency independently of its role in mRNA surveillance, which relies on Upf1p-Upf2p-Upf3p functional interplay. PMID- 14763986 TI - Topological analysis of Hin-catalysed DNA recombination in vivo and in vitro. AB - In vitro studies have demonstrated that Hin-catalysed site-specific DNA inversion occurs within a tripartite invertasome complex assembled at a branch on a supercoiled DNA molecule. Multiple DNA exchanges within a recombination complex (processive recombination) have been found to occur with particular substrates or reaction conditions. To investigate the mechanistic properties of the Hin recombination reaction in vivo, we have analysed the topology of recombination products generated by Hin catalysis in growing cells. Recombination between wild type recombination sites in vivo is primarily limited to one exchange. However, processive recombination leading to knotted DNA products is efficient on substrates containing recombination sites with non-identical core nucleotides. Multiple exchanges are limited by a short DNA segment between the Fis-bound enhancer and closest recombination site and by the strength of Fis-Hin interactions, implying that the enhancer normally remains associated with the recombining complex throughout a single exchange reaction, but that release of the enhancer leads to multiple exchanges. This work confirms salient mechanistic aspects of the reaction in vivo and provides strong evidence for the propensity of plectonemically branched DNA in prokaryotic cells. We also demonstrated that a single DNA exchange resulting in inversion in vitro is accompanied by a loss of four negative supercoils. PMID- 14763987 TI - Analysis of HutP-dependent transcription antitermination in the Bacillus subtilis hut operon: identification of HutP binding sites on hut antiterminator RNA and the involvement of the N-terminus of HutP in binding of HutP to the antiterminator RNA. AB - We investigated HutP-dependent transcription antitermination of the Bacillus subtilis hut operon. In vitro transcription assays with the B. subtilissigmaA containing RNA polymerase indicated that HutP inhibits transcription termination at the internal terminator by binding to the antiterminator on hut mRNA in the presence of histidine. Ethylnitrosourea modification interference assays and mutational analyses of the interference sites showed that interaction of HutP with a region containing three UAG trinucleotide sequences, which is located on top of the antiterminator structure, is critical for hut antitermination in vivo. Results from kinetic analysis of binding of HutP to RNA containing various portions of the antiterminator sequences indicated that secondary structure is required for binding of HutP to the region containing three UAG triplets in the antiterminator. The in vivo HutP antiterminator activity was reduced by the mutations in the N-terminal region of HutP. The HutP variants with H4A, R7A, I9A and Q26A mutations exhibited reduced binding affinities to the antiterminator RNA in vitro. A 25-mer peptide consisting of amino acid residues 2-26 of HutP bound to the antiterminator RNA. These results indicated that the N-terminus of HutP is involved in binding of HutP to the antiterminator RNA. PMID- 14763988 TI - Peptidoglycan hydrolytic activities associated with bacteriophage virions. AB - Murein hydrolases appear to be widespread in the virions of bacteriophages infecting Gram-positive or Gram-negative bacteria. Muralytic activity has been found in virions of the majority of a diverse collection of phages. Where known, the enzyme is either part of a large protein or found associated with other structural components of the virion that limit enzyme activity. In most cases, the lack of genetic and structural characterization of the phage precludes making a definitive identification of the enzymatic protein species. However, three proteins with muralytic activity have been unequivocally identified. T7gp16 is a 144 kDa internal head protein that is ejected into the cell at the initiation of infection; its enzyme activity is required only when the cell wall is more highly cross-linked. P22gp4 is part of the neck of the particle and is essential for infectivity. The activity associated with virions of Bacillus subtilis phage o29 and its relatives lies in the terminal protein gp3. These studies lead to a general mechanism describing how phage genomes are transported across the bacterial cell wall. PMID- 14763989 TI - Intein harbouring large tandem repeats in replicative DNA helicase of Trichodesmium erythraeum. AB - Protein splicing inteins can be small as approximately 130 aa or up to approximately 600 aa when harbouring an endonuclease domain. Here we report the identification and characterization of an unusually large intein, 1650 aa long and the largest of known inteins, encoded by the replicative DNA helicase gene dnaB of the oceanic N2-fixing cyanobacterium Trichodesmium erythraeum. This Ter DnaB-1 intein co-exists with a 177-aa mini-intein in the same host protein and harbours large tandem repeats in which an 84-aa sequence is repeated 16 times. Comparison between this tandem repeats and the recently reported tandem repeats of Ter DnaE-1 intein revealed differences and similarities. The two tandem repeats, residing in different inteins of different host proteins, differ by 50% in size and have little sequence similarity. Tandem repeats in the Ter DnaB-1 intein were required for the protein splicing activity when tested in Escherichia coli, in contrast to tandem repeats of the Ter DnaE-1 intein that inhibited protein splicing. On the other hand, tandem repeats of both inteins are located in the same corresponding region of the intein sequence and have the same number of repeating units. These suggest that the two tandem repeats could be related but have diverged greatly in size, sequence and effect on protein splicing. Alternatively, they could have independent origins but evolved certain similarities because of common constraints in structure and maintenance. PMID- 14763990 TI - Differences in two Pseudomonas aeruginosa cbb3 cytochrome oxidases. AB - Bacterial cytochrome cbb3 oxidases are members of the haeme-copper oxidase superfamily that are important for energy conservation by a variety of proteobacteria under oxygen-limiting conditions. The opportunistic pathogen Pseudomonas aeruginosa is unusual in possessing two operons that each potentially encode a cbb3 oxidase (cbb3-1 or cbb3-2). Our results demonstrate that, unlike typical enzymes of this class, the cbb3-1 oxidase has an important metabolic function at high oxygen tensions. In highly aerated cultures, cbb3-1 abundance and expression were greater than that of cbb3-2, and only loss of cbb3-1 influenced growth. Also, the activity of cbb3-1, not cbb3-2, inhibited expression of the alternative oxidase CioAB and thus influenced a signal transduction pathway much like that found in the alpha-proteobacterium Rhodobacter sphaeroides. Cbb3-2 appeared to play a more significant role under oxygen limitation by nature of its increased abundance and expression compared to highly aerated cultures, and the regulation of the cbb3-2 operon by the putative iron sulphur protein Anr. These results indicate that each of the two P. aeruginosa cbb3 isoforms have assumed specialized energetic and regulatory roles. PMID- 14763991 TI - The function of RNase G in Escherichia coli is constrained by its amino and carboxyl termini. AB - RNase G is a homologue of the essential Escherichia coli ribonuclease RNase E. Whereas RNase E plays a key role in the degradation of mRNA and the processing of tRNA and rRNA in E. coli, the biological functions of RNase G appear more limited. We report here that this difference in function is not merely a consequence of the significantly lower cellular concentration of RNase G, but also reflects differences in the intrinsic properties of these ribonucleases, as overproducing wild-type RNase G at a level up to 20 times the usual cellular concentration of RNase E cannot normally compensate for the absence of RNase E in E. coli. Instead, RNase G can sustain significant growth of RNase E-deficient E. coli cells only when it bears an unnatural extension at its amino terminus (e.g. MRKGINM) or carboxyl terminus (e.g. GHHHHHH). These extensions presumably enable RNase G to cleave critically important cellular RNAs whose efficient processing or degradation ordinarily requires RNase E. That extending the amino terminus of RNase G restores growth to E. coli cells lacking RNase E without detectably improving tRNA processing suggests that RNase E is not essential for tRNA production and is required for cell growth because it plays an indispensable role in the maturation or decay of essential E. coli RNAs other than tRNA. PMID- 14763993 TI - The controversy over levels of sex steroids in cases with Alzheimer's disease. PMID- 14763994 TI - Serum testosterone levels in males with Alzheimer's disease. AB - This study aimed to investigate whether there are differences in serum testosterone levels between male patients with Alzheimer's disease (AD) and cognitively normal male controls. Testosterone and sex hormone binding globulin (SHBG) levels were measured from 14 patients with mild to moderate AD and 16 age matched control males. The AD patients had higher levels of serum total (P = 0.02) and free testosterone (P < 0.001), and higher free androgen index (FAI) (P = 0.02) compared to controls. No differences were found for the SHBG levels. These data provide no support for hypotheses of (disproportionally) decreased levels of serum testosterone in AD. These data also show that all cognitively normal controls had an FAI below the normal range. PMID- 14763995 TI - p-Nonylphenol, 4-tert-octylphenol and bisphenol A increase the expression of progesterone receptor mRNA in the frontal cortex of adult ovariectomized rats. AB - Alkylphenols, such as p-nonylphenol (NP) and 4-tert-octylphenol (OP) and bisphenol A (BPA) are thought to mimic oestrogens in their action, and are called endocrine disrupters. We examined whether these endocrine disrupters affected progesterone receptor (PR) mRNA expression in the adult female rat neocortex. In one experiment, at 12.00 h, ovariectomized rats were given a subcutaneous injection of 10 mg of NP, 10 mg of OP or 10 mg of BPA, or sesame oil alone as control. Twenty-four hours after injection, the left side of the frontal cortex, parietal cortex and temporal cortex was collected. In a second experiment to study the time-course of the effects of BPA on PR mRNA, the ovariectomized rats were given a subcutaneous injection of 10 mg of BPA and killed 0, 6, 12 and 24 h after injection. In addition to the frontal cortex and temporal cortex, the occipital cortex was also collected. Northern blotting revealed that, in the first experiment, injection of NP, OP or BPA significantly increased PR mRNA expression in the frontal cortex but not in the parietal cortex. In the temporal cortex, BPA significantly decreased PR mRNA, but NP and OP produced no significant changes. The second experiment revealed that, in the frontal cortex, BPA induced a significant increase in PR mRNA expression at 6 h after injection, which lasted until 24 h after injection. In the temporal cortex, PR mRNA expression was significantly decreased 6 h after injection of BPA and was still significantly low 24 h after injection. No significant change was observed in the occipital cortex. These results suggest that, even in adult rats, endocrine disrupters alter the neocortical function by affecting the PR system, although the physiological significance of PR in the affected area is unknown. PMID- 14763996 TI - Fasting-induced activation of mitogen-activated protein kinases (ERK/p38) in the mouse hypothalamus. AB - Activity-dependent changes in neuronal plasticity depend critically on gene regulation. To understand how fasting-induced stimulation leads to gene regulation through intracellular signalling pathways, we investigated the effect of fasting on activation of the mitogen-activated protein kinase (MAPK) family, the extracellular signal-regulated kinase (ERK) and the p38 MAPK (p38) in mouse hypothalamus. In the hypothalamic arcuate nucleus, phosphorylation of ERK significantly increased during fasting, spatially coincident with phosphorylation of cAMP response element binding protein (CREB), induction of c-Fos, and expression of neuropeptide Y (NPY). In the paraventricular nucleus (PVN) of fasted mice, activation of p38 in addition to ERK was also observed. In the arcuate nucleus of ob/ob mice, phosphorylations of ERK and CREB were decreased during fasting, whereas the expression of NPY was increased. In the PVN, increased activation of p38 was observed in spite of decreased activation of ERK. These results suggest that ERK and p38 are differentially activated by fasting in distinct regions of the hypothalamus depending on the condition of energy balance. PMID- 14763997 TI - Interleukin-1beta stimulates growth hormone-releasing hormone receptor mRNA expression in the rat hypothalamus in vitro and in vivo. AB - Changes in growth hormone-releasing hormone (GHRH), GHRH-receptor (R), somatostatin and interleukin (IL)-1beta mRNA levels were determined in fetal rat hypothalamic cultures after administration of IL-1beta (1, 10, 100 ng/ml, 2 h incubation), and in adult rat hypothalamus 5 h after intracerebroventricular injection of IL-1beta (2.5 and 25 ng). IL-1beta stimulated GHRH-R mRNA expression both in vitro (10 and 100 ng/ml) and in vivo (2.5 and 25 ng). Somatostatin mRNA was significantly stimulated and GHRH mRNA slightly reduced in vitro, while these mRNA species were not altered in vivo in response to IL-1beta. IL-1beta stimulated its own expression both in vitro (10 and 100 ng/ml) and in vivo (25 ng). IL-1beta-induced mRNA responses occurred 2 h after treatment in vitro (incubation times, 30 min to 6 h). IL-1beta also elicited slight GHRH releases in vitro. Up-regulation of hypothalamic GHRH-R by IL-1beta may explain previous findings suggesting that IL-1beta stimulates GHRH activity. PMID- 14763998 TI - Activation of postsynaptic GABAB receptors modulate the firing activity of supraoptic oxytocin and vasopressin neurones: role of calcium channels. AB - Oxytocin and vasopressin release from neurohypophysial terminals is closely related to the firing activity of magnocellular neurones in the supraoptic (SON) and paraventricular nuclei. It is well established that activation of GABAA receptors potently inhibits the activity of SON neurones and, thus, hormone release. However, whether postsynaptic GABAB receptors are expressed in magnocellular neurones, and the role they play in controlling their firing activity, is still controversial. In the present work, we combined immunohistochemical and electrophysiological techniques to determine whether activation of GABAB receptors in identified oxytocin and vasopressin neurones modulates their firing activity. Patch-clamp recordings from SON neurones were obtained either in the slice preparation or from acutely dissociated neurones. Activation of GABAB receptors with the selective agonist baclofen (10 micro m) inhibited voltage-gated Ca2+ currents, reduced the duration of individual action potentials, as well as the magnitude of the hyperpolarizing after-potential. SON firing activity was reduced by baclofen, and effect that was accompanied by a small membrane hyperpolarization. The inhibition of firing discharge persisted in the presence of synaptic blockade media, and was also observed in acutely dissociated SON neurones. Finally, GABAB-mediated modulation of firing activity was largely blocked by the Ca2+ channel blocker Co2+ (2 mm). In general, baclofen modulatory actions were significantly larger, or observed more predominantly, in vasopressin neurones. In summary, these results support the expression of functional postsynaptic GABAB receptors in SON neurones, activation of which efficiently modulates neuronal excitability, in a Ca2+- and cell-type dependent manner. PMID- 14763999 TI - Responses of cells in the rat supraoptic nucleus in vivo to stimulation of afferent pathways are different at different times of the light/dark cycle. AB - To determine whether the daily rhythms of spike activity in the supraoptic nucleus (SON) were accompanied by changes in the behaviour of its inputs, we used conventional extracellular single cell recordings from cells in the SON of anaesthetized rats while stimulating the contralateral optic nerve and the ipsilateral suprachiasmatic nucleus (SCN). Neurones in the SON region were identified by antidromic activation and classified as oxytocin or vasopressin cells, on the basis of their spontaneous firing patterns. Approximately 27% of both oxytocin (29/108) and vasopressin (39/147) neurones were excited by stimulation of the optic nerve, and the majority of responses had a long latency (>20 ms). Very few oxytocin (3/108) and vasopressin cells (2/147) were inhibited by stimulation of the optic nerve. The pattern of the responses (excitatory, inhibitory or nonresponsive) of oxytocin and vasopressin cells to stimulation of the optic nerve was significantly related to the time of day (chi-square test; P = 0.012, oxytocin cells; P = 0.006, vasopressin cells). The proportion of oxytocin cells excited by stimulation of the optic nerve was highest at ZT 4-8 and lowest at ZT 20-24. For vasopressin cells, it was highest at ZT 12-16 and lowest at ZT 20-24. The proportion of excitatory, inhibitory and complex responses seen in oxytocin and vasopressin cells following stimulation of the SCN also changed and was significantly different at different times of day (oxytocin cells: highest proportion of excitatory responses at ZT 12-16, P = 0.029; chi square test; vasopressin cells: highest proportion of excitatory responses at ZT 0-4, P = 0.005; chi-square test). Thus, inputs to oxytocin and vasopressin neurones from the optic nerve and some outputs from the SCN changed during the light/dark cycle. Such changes may contribute to the generation of 24-h rhythms in activity of oxytocin and vasopressin neurones and release of the peptides. PMID- 14764000 TI - Regulation of galanin receptor GalR1 mRNA expression by ovarian steroids in oestrogen receptor alpha-immunoreactive neurones: identification of distinct populations of neurones in the preoptic area. AB - This study examined whether gonadal steroids are involved in regulating galanin receptor 1 (GalR1) mRNA expression in neurones that contain oestrogen receptor alpha (ERalpha), in three regions of the preoptic area (POA) known to be involved in the control of gonadotropin secretion. Double-labelling immunohistochemistry using an antibody against the ERalpha and in situ hybridization experiments using a 35S-labelled riboprobe specific for GalR1 mRNAs revealed that ERalpha is expressed in a large proportion of GalR1 mRNA-expressing neurones of the POA in the ovariectomized (OVX) female rat. Oestradiol (E2) and oestradiol plus progesterone (E2 + P) treatments of OVX rats significantly decreased the proportion of GalR1 mRNA/ERalpha immunoreactive (ERalpha-IR) neurones in the anteroventral periventricular nucleus (AVPV), medial preoptic nucleus (MPN) and medial preoptic area (MPO). The expression of GalR1 mRNA in ERalpha-IR neurones varied according the hormonal status of the female animals. In the AVPV, during the oestrous cycle, the hybridization signal significantly increased at oestrus. E2 and E2 + P treatments of OVX rats did not induced significant variation of levels of GalR1 mRNAs in ERalpha-IR neurones. In the MPN, E2 treatment of OVX rats resulted in significant increase in GalR1 mRNA expression in ERalpha-IR neurones. Similarly, levels of the GalR1 hybridization signal increased during afternoon of proestrus and oestrus. In the MPO, treatment of OVX rats with E2 + P significantly decreased GalR1 mRNA expression in ERalpha-IR neurones. The expression of GalR1 mRNA did not change during the oestrous cycle in this area. These findings suggest that the hypothalamic action of galanin on gonadotopin releasing hormone (GnRH) secretion may pass through the specific population of GalR1/ERalpha-IR neurones of the MPN in mediating the oestrogen action on the GnRH system at the moment of the luteinizing hormone surge. PMID- 14764001 TI - Effects of ovariectomy on the neuroendocrine axes regulating reproduction and energy balance in young cynomolgus macaques. AB - Degeneration of the ovary in middle-aged women results in castrate levels of ovarian steroids and increased gonadotropin secretion from the anterior pituitary gland. Ageing in women is also accompanied by significant changes in energy homeostasis. We have observed alterations in hypothalamic morphology and gene expression in older women, including hypertrophy and increased gene expression of neurokinin B (NKB) neurones, elevated levels of gonadotropin releasing-hormone (GnRH) mRNA and decreased numbers of neurones expressing pro-opiomelanocortin (POMC) mRNA. To determine if loss of ovarian steroids could produce comparable changes in gene expression in young primates, we measured the effects of ovariectomy on NKB, GnRH and POMC gene expression in young cynomolgus monkeys. We also measured serum leptin and body weight to examine the consequences of ovariectomy on energy balance. NKB neurones in the infundibular nucleus of ovariectomized monkeys were larger, more numerous and displayed increased levels of NKB mRNA compared to those of intact controls. Moreover, ovariectomy increased the number of neurones expressing GnRH gene transcripts and elevated serum luteinizing hormone. By contrast, several parameters related to energy balance, including POMC gene expression, serum leptin and body weights, were unchanged by ovariectomy. Thus, the rise in NKB and GnRH gene expression in older women was simulated by ovariectomy in monkeys, but the changes in POMC gene expression and energy balance were not. This study provides strong support for the hypothesis that ovarian failure contributes to the increased NKB and GnRH gene expression observed in postmenopausal women. PMID- 14764002 TI - Hormone treatment facilitates penile erection in castrated rats after sleep deprivation and cocaine. AB - Sleep deprivation is associated with cocaine-enhanced genital reflexes in male rats, and castration of the male rat causes a decline in sexual behaviour, which can be reversed by hormone administration. We conducted two experiments to determine whether sleep deprivation and cocaine administration could also induce spontaneous penile erection in castrated rats after hormonal treatment (testosterone, progesterone and oestradiol). Different doses of hormones or vehicle were administered to rats during the 4-day period of sleep deprivation, and in home-cage control rats. Testosterone did not restore penile erection in castrated sleep-deprived rats. Progesterone triggered penile erection, and 100 mg/day of progesterone induced the highest proportion of rats displaying penile erection, and restored the frequency of penile erection observed in noncastrated sleep deprived rats. Penile erection was absent in vehicle as well as oestradiol treated sleep-deprived castrated rats. Whereas sleep deprivation increased progesterone concentrations in noncastrated rats, sleep deprivation decreased progesterone concentrations in castrated rats. Corticosterone concentrations were lower in the castrated sleep-deprived rats than in respective control group. These data show that progesterone treatment facilitates penile erection in sleep deprived-cocaine castrated rats. PMID- 14764003 TI - The hypothalamic insulin-like growth factor-1 receptor and its relationship to gonadotropin-releasing hormones neurones during postnatal development. AB - Reproduction in vertebrates is controlled by hypophysiotropic gonadotropin releasing hormone (GnRH) neurones. Pulsatile GnRH release increases during reproductive development, resulting in the onset and progression of puberty and, ultimately, the acquisition and maintenance of adult reproductive function. These changes in GnRH release are largely due to inputs to GnRH cells from other factors, including the neurotrophic factor, insulin-like growth factor-1 (IGF-1). Here, molecular studies were undertaken to quantify expression of IGF-1 receptor (IGF-1R) mRNA in the preoptic area-anterior hypothalamus (POA-AH) and mediobasal hypothalamus (MBH)-median eminence (ME), the sites of GnRH perikarya and neuroterminals, respectively. Immunocytochemical studies were also carried out to study the anatomical relationship between the IGF-1R and GnRH neurones. Experiments were performed in a developmental context using neonatal (P5), peripubertal ( approximately P30) and adult (P60) male and female mice. We found that IGF-1R mRNA levels in the POA-AH were significantly different among all age groups, with levels higher at P60 then P5 or approximately P30. Levels of IGF-1R mRNA in the MBH-ME were lower at P5 than approximately P30 or P60. Qualitative observations suggested that IGF-1R immunoreactivity in POA-AH increased from P5 through P60. Quantitative double-label immunocytochemistry studies showed that GnRH perikarya expressed IGF-1R. Taken together, the results demonstrate expression of, and developmental changes in, IGF-1R gene and protein in brain regions containing GnRH and other neuroendocrine cells. Moreover, the novel finding that the IGF-1R is expressed on GnRH perikarya in vivo suggests a potential direct anatomical locus where IGF-1 can regulate reproductive development and function. PMID- 14764004 TI - Single chain variants of the glycoprotein hormones and their receptors as tools to study receptor activation and for analogue design. AB - One of the crucial steps in the biosynthesis of multisubunit proteins is their assembly. The glycoprotein hormone, thyroid-stimulating hormone, and the gonadotropins, luteinizing hormone, follicle-stimulating hormone and chorionic gonadotropin, are noncovalent heterodimers. Their assembly is critical for bioactivity because the heterodimers, but not the monomeric subunits, efficiently bind to and activate the cognate heptahelical receptor. Occasionally, mutated subunits cannot combine into a functional hormone, or the bioactivity of the assembled, yet modified, heterodimer is suboptimal. PMID- 14764005 TI - Seeing the light...in a new way. AB - Research over the past decade has shown that the rods and cones are not the only photoreceptors of the eye. There also exists a population of directly light sensitive ganglion cells that act as brightness detectors and regulate a wide variety of different photosensory tasks, including the regulation of 24-h circadian clocks, synthesis of the hormone melatonin, pupil size and behaviour. These receptors may even help modulate our mood and sense of well-being. PMID- 14764006 TI - Cultural sensitivity: the soft option? PMID- 14764007 TI - Cultural, behaviour and HIV/AIDS. PMID- 14764008 TI - Nurses in Zambia gain access to antiretroviral treatment. PMID- 14764009 TI - The battle against AIDS needs all of us. PMID- 14764010 TI - Adolescents' views on decision-making regarding risky sexual behaviour. AB - PURPOSE: To explore adolescents' views regarding risky sexual behaviour who were aged between 13 and 19 years. METHODS: A purposive sample was drawn from adolescents who were participating in workshops, organized by the Ministry of Health and Social Welfare. The sample consisted of a mix of in-school and out-of school adolescents aged between 13 and 19 years. The overall aim of the workshops was to explore adolescents' views on decision-making regarding risky sexual behaviour. The educational status of participants lay between grades 0 and 12. A total of 24 focus group discussions were conducted, six in each of the four regions of Swaziland (Manzini, Hhohho, Lubombo and Shiselweni). Each group consisted of 6-12 participants. All focus group discussions were audiotaped and later transcribed. FINDINGS: The following four major themes emerged from content analysis: sexual behaviour, age at first intercourse, peer pressure and trust in the relationship. These were all found to be important factors in decision-making for risky sexual behaviours. CONCLUSIONS: Obtaining a sexual history, motivation to use condoms, guidance and counselling on ways to reduce adolescents' risk of sexual exploitation, and peer education programmes, are all of importance when aiming to reduce risky sexual behaviour. RECOMMENDATION: Adolescents should receive positive reinforcement for responsible sexual behaviour including abstinence and, where appropriate, use of active birth control. PMID- 14764011 TI - A framework for short-term humanitarian health care projects. AB - BACKGROUND: This opinion piece, based on the experiences of the author, outlines the planning involved for a short-term assignment. With growing demand for humanitarian health care assistance to underserved countries, a framework for organizing short-term trips to such areas would be beneficial to the trip organizers. CONTENT: The key to a successful project lies in focusing on specific populations and interventions while still in the planning stages. The hurdles that need to be cleared before actually embarking on the project are also discussed. OUTCOMES: The lasting result of such ventures lies in the learning experiences of the participants, the care that the population receives, and a deeper appreciation for global health and disparity. PMID- 14764012 TI - Exploring the health care status of two communities in the Dominican Republic. AB - BACKGROUND: This article looks at two impoverished communities in the Dominican Republic that are working in collaboration with a USA-based organization to meet the physical, emotional, educational, and spiritual needs of the children in these areas. METHODS: An exploratory descriptive study was performed to establish baseline information. The knowledge gained from this study will provide guidance in the development of effective school-based strategies to meet the physical, emotional, and nutritional needs of the children. Data gathered from health records obtained during medical clinics in these two communities were reviewed. Descriptive statistics were performed to examine demographics, chief complaints, diagnoses, and treatments of approximately 1500 Dominicans. FINDINGS: Results support that living conditions and the lack of primary care continue to underpin the majority of health issues within these communities. OUTCOMES: School-based health initiatives along with a well child-screening programme are being developed to enhance the health of the children in these communities. PMID- 14764013 TI - Behavioural, emotional and family functioning of hospitalized children in China and Hong Kong. AB - PURPOSE: This study examined behavioural and emotional problems, social competence and family functioning of hospitalized Chinese children in Hong Kong and the Chinese Mainland. METHOD: A sample of 210 hospitalized children (ages 2 11 years) and their families participated in the study. The families were from a cross-section of geographical areas in Hong Kong (two hospitals) and the Chinese Mainland (five hospitals). Parents completed an age-appropriate Chinese version of the Child Behaviour Checklist and the Family Assessment Device. Multiple regression models were used to examine predictors of children's behaviour problems. RESULTS: Behavioural patterns appeared to be specific to the developmental stage. Children had greater problems when their families demonstrated poorer affective involvement. Hospitalized children on the Chinese Mainland experienced more internalizing and externalizing behaviour problems than those in Hong Kong. Sick children, according to their parents, however, demonstrate some resiliency based on social and academic competency factors. CONCLUSIONS: Hospitalized Chinese children manifest behavioural, emotional and family problems that vary by region, the child's development and gender. Problems predominantly of an internalizing nature characterized this group. The findings support the need for culturally appropriate behavioural assessments and interventions with hospitalized children. PMID- 14764014 TI - A review of the literature on care of clients in pain who are culturally diverse. AB - AIM: This article reviews the literature on the care of clients from diverse cultures who are in pain and provides strategies for care. BACKGROUND: Pain is a critical concept for caring for clients and particularly for clients from another culture. Culture shapes the values, beliefs, norms, and practices of individuals, including the ways persons react to pain. Culture affects the assessment and management of pain. CONCLUSIONS: Seven strategies can assist in culturally appropriate assessment and management of pain: (1) utilize assessment tools to assist in measuring pain, (2) appreciate variations in affective response to pain, (3) be sensitive to variations in communication styles, (4) recognize that communication of pain may not be acceptable within a culture, (5) appreciate that the meaning of pain varies between cultures, (6) utilize knowledge of biological variations, and (7) develop personal awareness of values and beliefs which may affect responses to pain. PMID- 14764015 TI - Effects of acupuncture and care interventions on the outcome of inflammatory symptoms of the breast in lactating women. AB - OBJECTIVES: To compare modes of care and treatment for lactating women with inflammatory symptoms of the breast, with special focus on the use of acupuncture. METHOD: Eighty-eight mothers were randomized into three treatment groups. All three groups were given advice regarding emptying of the breasts and care in the form of comfort interventions. Acupuncture was included in the treatment regime for two of the groups. A severity index was created by adding together scores for signs and symptoms: breast tension, erythema and pain. FINDINGS: Mothers in all groups expressed relative satisfaction with the breastfeeding situation despite considerable discomfort. There was no significant difference between the groups for the number of mothers requiring more than three contact days for recovery nor for their severity index scores on day 3. These findings must be interpreted with care but may suggest that care interventions play as great a part in the recovery of these women as acupuncture treatment or the use of oxytocin spray. Antibiotic therapy was used in 9% of the study population, which is in contrast to other studies. PROPOSED FUTURE ACTION: Based on these results, a new study has been designed to test the hypothesis that acupuncture hastens recovery from inflammatory processes in the lactating breast and approximately 200 mothers will be randomized in a new expanded study. PMID- 14764016 TI - Horses for courses. PMID- 14764017 TI - A systematic review on the effectiveness of alcohol-based solutions for hand hygiene. AB - The use of alcohol has been proposed as an option for hand hygiene. A systematic review was conducted to evaluate the clinical evidence supporting the use of alcohol-based solutions in hospitals as an option for hand hygiene. Studies published between January 1992 and April 2002 in English and Thai, related to the effectiveness of alcohol-based solutions, were reviewed. The databases searched included Medline, DARE, CINAHL and Dissertation Abstracts International. All studies were assessed as having adequate methodological quality. Results of this systematic review supported that alcohol-based hand rubbing removes microorganisms effectively, requires less time and irritates hands less often than does handwashing with soap or other antiseptic agents and water. Furthermore, the availability of bedside alcohol-based solutions increases compliance with hand hygiene among health care workers. PMID- 14764018 TI - Part two: The core components of legitimate influence and the conditions that constrain or facilitate advanced nursing practice in adult critical care. AB - This paper describes intervening conditions that might constrain or facilitate the exercise of Legitimate Influence: The Key to Advanced Nursing in Adult Critical Care, the foundation of which is credibility and advanced clinical nursing practice. Constraining conditions are conflict, resistance, gender bias, political awareness and established values. Credibility, advanced clinical nursing practice and strategic activity are required to enhance patient stay in hospital and improve patient outcome. Intervening conditions that facilitate these are overcoming resistance, political awareness and established values. In a previous paper, it was indicated that enhanced patient stay and improved patient outcome were achieved primarily through strategic activity that emphasized restoring patients to a former, or improved, health status. This paper portrays how intervening conditions can impinge upon this and the exercise of legitimate influence. PMID- 14764019 TI - Who, when and where? Identification of patients at risk of an in-hospital adverse event: implications for nursing practice. AB - Patients who suffer an adverse event (AE) are more likely to die or suffer permanent disability. Many AEs are preventable. Nurses have long played a pivotal role in the prevention of AEs. Much of the literature to date pertains to the role of nurses in the prevention of AEs such as falls, pressure areas and deep vein thrombosis. Prominent risk factors for AEs are the presence of physiological abnormality, failure to recognize or correct physiological abnormality, advanced patient age and location of patient room. Ongoing physiological assessment of patients is a nursing responsibility and the assessment findings of nurses underpin many patient care decisions. The early recognition and correction of physiological abnormality can improve patient outcomes by reducing the incidence of AEs, making nurses' ability to identify, interpret and act on physiological abnormality a fundamental factor in AE prediction and prevention. This paper will examine the role of nurses in AE prevention, using cardiac arrest as an example, from the perspective of physiological safety; that is, accurate physiological assessment and the early correction of physiological abnormality. PMID- 14764020 TI - The attitudes, concerns, gloving practices and knowledge of nurses in a Taiwanese hospital regarding AIDS and HIV. AB - Many studies have documented nurses' attitudes and concerns regarding AIDS/HIV, but little is known about Taiwanese nurses. We documented attitudes, concerns, gloving practices and practical AIDS/HIV knowledge of 1090 nurses from one metropolitan hospital in Changhua City, Taiwan. The response rate was 80.9%. Both HIV and hepatitis contraction in the workplace was nurses' main concern. Two hundred and ten nurses (19.3%) were seriously considering leaving nursing because of fear of contracting AIDS/HIV. Virtually all nurses considered it their right to be informed of the presence of HIV-positive patients in their direct work area and many believed that HIV testing of patients should be mandatory. Practical AIDS/HIV knowledge was deficient. These Taiwanese nurses have concerns and fears that might be related to deficiencies in practical AIDS/HIV knowledge. Continuous educational programmes are recommended to alleviate these nurses' attitudes and concerns regarding AIDS/HIV. PMID- 14764021 TI - The context, measures and outcomes of psychosocial care interventions in long term health care for older people. AB - This paper examines the psychosocial dimensions of long-term care with reference to the new International Classification of Functioning, Disability and Handicap (ICIDH 2) and to research conducted in Hong Kong. It also draws on selected international literature about older people. It discusses the different ways in which information can be gained about the personal, social and emotional processes of rehabilitation that influence outcomes and raises methodological questions about the study of interventions. Outcomes that are sensitive to psychosocial interventions and that take account of the elderly person's own perspective are identified as important challenges for nurses and other professionals in the multidisciplinary team, in order to respond to an individualized approach to long-term care. It is concluded that gaining a better understanding of the psychosocial dimensions of long-term care will enhance professional practice and benefit older people and their carers. PMID- 14764022 TI - The life situation and functional capacity of the elderly with locomotor disability in Sweden and Poland according to a model by Lawton. AB - A quadripartite concept, the Good Life for older people, was developed by the American psychologist M. P. Lawton and forms the theoretical framework of this study. Ninety Swedish and 93 Polish subjects, aged > or = 60 years who had reported locomotor disturbances in selected samples from the two countries, took part in the study. Interviews were performed using the Philadelphia Geriatric Center Multilevel Assessment Instrument (PGCMAI) and functional testing was done using the Standardized Practical Equipment (SPE). The Polish elderly scored lower in most domains of the PGCMAI, which meant a worse life situation according to the Good Life model. On the SPE, scoring was lower in the Polish group, mainly on items related to balance and mobility. There was a logic convergent validity between the PGCMAI and the SPE in the whole group. The somewhat more complicated life situation for the Polish elderly has to be further analysed. PMID- 14764023 TI - Networking for the advancement of nursing research in Europe for twenty-five years. AB - The Work Group of European Nurse Researchers (WENR) has been networking for advancement of nursing research in Europe for 25 years. The most well-known activity of WENR is its biennial conferences organized in all parts of Europe. They have provided a unique opportunity for nurses to come together to discuss nursing research and clinical practice. Recently, WENR has published a position paper in order to reinforce its own and International Council of Nurses, World Health Organization and The Standing Committee of Nurses of the European Union (PCN) aims. The Work Group of European Nurse Researchers recommends that the recommendations of The Council of Europe report be adopted formally by national nurses associations. Future activities of WENR include promotion of infrastructures and research capacities in member countries, and planning of new scientific activities. PMID- 14764024 TI - The exploding worldwide cancer burden: the impact of cancer on women. AB - Although age-adjusted cancer death rates have started to decline in the United States and other developed nations - thanks in large part to widespread screening programs that detect cancers at early, treatable stages - cancer in developing countries is on the rise. Ironically, rising life expectancy in those nations along with the adoption of 'Western' lifestyles will leave many more people vulnerable to cancer. Unfortunately, the early detection tools and treatment technology that have helped control cancer in wealthier lands are often not readily available in many other countries. Much of this increased cancer burden will take the form of cancers that affect women - not only breast, cervical, and other gynecologic cancers but colorectal cancer, lung cancer, and other malignancies related to tobacco. Physicians specializing in cancer care for women need to be alert to every opportunity to improve cancer screening and prevention among the growing, aging populations of less-developed countries. Less precise but less costly and more widely available screening techniques may save thousands more lives than the most sophisticated technology because low-cost programs can be applied widely instead of being reserved for a fortunate few. In addition, education and prevention efforts directed toward tobacco use need to be put in place to help stem an epidemic of tobacco-related cancers that has largely peaked in developed countries but looms ominously in the future of developing nations. PMID- 14764026 TI - The relative importance of surgical training and laboratory research in a gynecologic oncology fellowship. AB - The objective was to determine trends of surgical skill acquisition during fellowships, and the consensus amongst gynecologic oncologists about the relative importance of surgical training and laboratory research in fellowships. A survey addressing surgical capability at the time of fellowship completion, and relative priorities that should be given to surgical training and laboratory research was mailed to gynecologic oncologists and fellows in the Society of Gynecologic Oncologists directory. Of 820 surveyed, 454 (55.4%) of provided utilizable data, of whom 56 (12.5%) were fellows, and 398 (87.5%) in practice (49.5% university based and 50.5% community hospital-based). Relative to past graduates, recent ones report and current fellows anticipate a lower probability of being able to independently perform some procedures applicable to cervical and ovarian cancer, as well as others necessary to manage complications at the time of fellowship completion. 69.8% of all respondents think that greater emphasis should be placed on surgical training at the expense of doing less laboratory research. There is wide variation of opinion among respondents concerning the value of and most appropriate length of time that should be dedicated to laboratory research in a fellowship. There is an indication of a trend for more recent fellows to graduate having acquired less surgical skill and a prevalent opinion that surgical training should be more heavily emphasized in fellowships. PMID- 14764025 TI - Whole-body positron emission tomography and positron emission tomography/computed tomography in gynecologic oncology. AB - The advent of positron emission tomography (PET) and PET/computed tomography (CT) now enables us to detect metabolically active gynecologic cancers with greater accuracy than was possible with anatomic imagings. Fluorine-18 fluorodeoxyglucose PET has been useful in differentiation of malignant from benign lymph nodes, and residual or recurrent cancers from post-treatment changes. PET/CT produces additional information for the diagnosis and tissue biopsy as well as radiotherapy planning. This paper reviews the principle and clinical utility of PET and PET/CT in the diagnosis, staging, recurrence, therapeutic response as well as prognosis of gynecologic cancers. PMID- 14764027 TI - Long-term results of cytoreductive surgery for advanced and recurrent epithelial ovarian cancers and papillary serous carcinoma of the peritoneum. AB - The aim of this study was to review the long-term results of cytoreductive surgery in the treatment of advanced primary and recurrent epithelial ovarian cancers and papillary serous carcinoma of the peritoneum. Our goal was to identify clinical factors by which to select patients likely to benefit from a comprehensive management plan. Clinical data of 28 females who underwent surgery were retrieved from a prospective database. Major cytoreductive procedures were possible in 25 patients. Heated intraoperative or early postoperative intraperitoneal chemotherapy was also used where appropriate. The median follow up after cytoreduction was 26.9 months. The overall median survival after cytoreduction was 45.8 months. The prognostic indicators associated with a statistically significant impact on survival were the prior surgery score (P < 0.001), the completeness of cytoreduction score (CC; P = 0.037), and response to chemotherapy prior to surgery (P = 0.012). Our findings suggest that cytoreductive surgery can be effective when combined with perioperative intraperitoneal chemotherapy. Results can be improved by excluding cases where CC seems unlikely. Extensive prior surgery without the protection of adjunctive intraperitoneal chemotherapy is associated with a poor prognosis. This may be due to disruption of anatomical planes leading to deep abdominal and pelvic dissemination intractable to further treatment. PMID- 14764028 TI - Preoperative predictors of suboptimal primary surgical cytoreduction in women with clinical evidence of advanced primary epithelial ovarian cancer. AB - INTRODUCTION: Women with epithelial ovarian cancer (EOC) are conventionally treated with primary cytoreductive surgery. For those with a low probability of optimal primary surgical debulking, an alternative management option is primary chemotherapy with delayed primary surgery. Selection criteria are required to identify women who may benefit from this approach. PATIENTS AND METHODS: Patient age, presence of ascites, preoperative serum CA-125 level, surgical procedures performed, postoperative residual disease, FIGO stage, and histology data were collected on 97 women with preoperative clinical evidence of advanced EOC. Univariate and multivariate analysis was performed to identify which preoperative factors predict disease that will be suboptimally debulked. Receiver-operator curves were constructed for CA-125 level as a predictor for residual disease. RESULTS: The best predictor of disease suboptimally cytoreduced was serum CA-125 level (OR = 22.76, 95% CI = 7.13-72.69). Other predictive factors included age over 60 years (OR = 3.16, 95% CI = 1.04-9.56) and clinical evidence of ascites (OR = 3.30, 95% CI = 1.03-10.62). The optimal cut-off for serum CA-125 as a predictor of suboptimal debulking was 586 IU (sensitivity 80.0%, specificity 88.5%, PPV 85.7%). CONCLUSION: Serum CA-125 level is a reliable component of the preoperative assessment of women with EOC. PMID- 14764029 TI - The influence of splenic metastases on survival in FIGO stage IIIC epithelial ovarian cancer. AB - The purposes of this study were to compare the survival of ovarian cancer patients with splenic metastasis to patients without it and to evaluate the complications of the procedure. A retrospective study was performed on 34 patients with ovarian cancer who underwent splenectomy for initial cytoreduction at gynecologic oncology unit of Hacettepe University Hospitals between 1989 and 2001. All patients had FIGO stage IIIC disease and were left with <1 cm residual tumor after surgery. Eighteen patients (52.9%) had splenic metastasis. Patients with splenic metastasis tended poorer survival. Median survivals were 28.9 and 41.3 months for patients with splenic disease and for patients without it, respectively (P > 0.05). Univariate analysis revealed that performance status and histologic type influenced survival. Histologic type and performance status were identified as independent risk factors by multivariate analysis. Postoperative complications were developed in ten (29.4%) patients and three of these (8.8%) died in 1 month after operation. None of the complications was attributed directly to the splenectomy procedure. Complete surgical cytoreduction confers a survival benefit whether the parenchyma was involved or not. The splenectomy should be considered with its acceptable morbidity in selected patients who have a chance to achieve optimal debulking. PMID- 14764031 TI - Restoration of tumor specific human leukocyte antigens class I-restricted cytotoxicity by dendritic cell stimulation of tumor infiltrating lymphocytes in patients with advanced ovarian cancer. AB - Despite the large number of potentially cytotoxic tumor-infiltrating (TIL) and tumor-associated (TAL) lymphocytes accumulated in the peritoneal cavity ascitic fluid and tumor tissue, advanced ovarian cancer is a progressive disease, suggesting that TIL and TAL populations eventually become functionally suppressed in vivo. Dendritic cells (DC) are the most powerful professional antigen presenting cells known in humans and recently, ovarian tumor antigen pulsed DC have been shown to elicit tumor specific human leukocyte antigens (HLA)-class I restricted cytotoxicity from the peripheral blood of advanced ovarian cancer patients. In this study, we have evaluated the potential of tumor antigen-pulsed fully mature DC stimulation in restoring tumor-specific cytotoxicity in anergic TIL populations from advanced ovarian cancer patients. In addition, we have compared tumor-specific T-cell responses induced by tumor antigen-loaded DC in TIL to those induced in TAL and peripheral blood lymphocytes (PBL). DC stimulation induced powerful cytotoxicity against autologous tumor target cells in TIL-derived CD8+ T-cells from all patients tested, while autologous Epstein Barr virus (EBV)-transformed lymphoblastoid cell lines (LCL) were not lysed. Killing of autologous tumor cells was higher by CD8+ T-cells from TIL compared to PBL and TAL (P < 0.01) and was more strongly inhibited by anti-HLA class I MAb (P < 0.05 compared to PBL and TAL). Phenotypically, all cytotoxic T lymphocyte (CTL) populations were CD3+/CD8+, with variable levels of CD56 expression. Finally, although a marked Type 1 cytokine bias [ie, interferon-gamma/interleukin-4 (IFN gammahigh/IL-4low)] was observable in all DC-stimulated CD8+ T-cell populations, TIL derived CD8+ T-cells showed a higher percentage of IFN-gamma positive cells compared to TAL and PBL. Taken together, these data show that tumor lysate-pulsed DC can consistently restore strong CD8+ CTL responses from TIL against autologous ovarian cancer cells. DC-stimulated TIL may represent a superior source of tumor specific CTL for adoptive T-cell immunotherapy for advanced ovarian cancer. PMID- 14764030 TI - Comparison of hydrosonography and transvaginal ultrasonography in the detection of intracavitary pathologies in women with abnormal uterine bleeding. AB - BACKGROUND: The aim of the study was to compare the accuracy of hydrosonography with that of transvaginal ultrasonography in detection of intracavitary pathologies in patients with history of abnormal uterine bleeding. STUDY DESIGN: Prospective, randomized, and unblinded study. MATERIAL AND METHODS: A total of 197 women (n = 130 premenopausal and n = 67 postmenopausal) aged between 23 and 71 years (mean age 45.7 +/- 8.9) presenting with a history of abnormal uterine bleeding were included into the study. Hydrosonography was carried out by experienced gynecologists, on the same setting in an outpatient clinic immediately after the performance of transvaginal sonography. The finally obtained surgical-pathologic findings were compared with the results obtained from transvaginal sonography and hydrosonography. Sensitivity, specificity, positive, and negative predictive values were calculated for each procedure. RESULTS: The surgical-pathologic examination confirmed normal physiologic endometrium in 50 (48%) of 104 women who were said to have normal endometrium on transvaginal sonography. Seventy (75%) of 93 women diagnosed of intracavitary pathologies on transvaginal sonography were confirmed by surgical-pathologic findings. The sensitivity, specificity, positive predictive value, and negative predictive value of transvaginal sonography in the detection of intracavitary pathology were 56, 68, 75, and 48%, respectively. Surgical-pathologic results revealed intracavitary pathologies in 23 (30%) of 76 women who were said to have normal endometrium on hydrosonography. Among 121 women diagnosed of intracavitary pathologies on hydrosonography, 101 (81%) women were confirmed after histological evaluation of the surgical specimens. The sensitivity, specificity, positive predictive value, and negative predictive value of hydrosonography in the detection of intracavitary pathology were 81, 73, 83, and 70%, respectively. Sensitivity and negative predictive value were significantly higher with hydrosonography. There were five cases of endometrial malignancy in which one of the case of malignancy was on polyp and two cases of endometrial hyperplasia with atypia which were not stated on sonographic results. CONCLUSION: Hydrosonography is more accurate than transvaginal ultrasography in the detection of intracavitary pathologies in women with abnormal uterine bleeding. PMID- 14764032 TI - Mechanisms of reversal of adriamycin resistance in human ovarian carcinoma cell line by ultrasound. AB - The aim of this study was to investigate the reversal of adriamycin resistance in human ovarian cancer cells by ultrasound exposure from perspectives of apoptosis and mdr1 gene. Apoptosis was determined by flow cytometry and mdr1 level by reverse transcription-polymerase chain reaction (RT-PCR). Apoptosis indexes were improved in groups in which ultrasound exposures were applied. RT-PCR did not support that insonation-decreased mdr1 level. These findings suggested that the ultrasound-enhanced therapeutic efficacy was not mediated via modulating gene expression, and ultrasound maybe lowered thresholds for apoptosis and oncosis in chemoresistant ovarian cancer cells. PMID- 14764033 TI - Prognostic significance of tumor angiogenesis in epithelial ovarian cancer: in association with transforming growth factor beta and vascular endothelial growth factor. AB - We aimed to evaluate the prognostic significance of microvessel density (MVD), vascular endothelial growth factor (VEGF), and transforming growth factor beta (TGFbeta), as well as to find out the relationship between MVD, and VEGF and TGFbeta in epithelial ovarian cancer (EOC). Surgical specimens of 47 patients with stage I-IV primary EOC, who underwent extended surgical staging according to FIGO, were investigated. Five- micro m thick tissue sections were immunostained with antibody to factor VIII-related antigen, and MVD was assessed at three separate areas of x200 magnification. Expressions for VEGF and TGFbeta were evaluated by immunohistochemical staining using related monoclonal antibodies. Results were correlated with clinicopathologic factors and survival. We did not find any correlation between MVD and clinicopathologic factors, or patient survival. Similarly, there was no association between the degree of VEGF staining and survival or clinicopathologic factors, except preoperative ascites volume, which was higher in patients showing moderate and intense VEGF staining than those with weak VEGF staining (P = 0.052). The expression of TGFbeta was inversely correlated with preoperative CA-125 levels (P < 0.05). Furthermore, there was no correlation between MVD and the staining intensity of VEGF or TGFbeta. In conclusion, angiogenesis does not appear as a prognostic factor in EOC. We suggest that VEGF is an important mediator of ascites formation, and that TGFbeta, which is supposed to have tissue-specific actions in tumorigenesis, may have growth-inhibitory functions in EOC. PMID- 14764034 TI - Sulfatides in ovarian tumors: clinicopathological correlates. AB - OBJECTIVES: To investigate the expression of sulfatides in the tissue homogenates of malignant ovarian tumors, benign ovarian tumors, and control tissues and to study the relation between this marker and other clinico-pathological criteria such as the tumor type, grade of differentiation, surgical stage and ovulatory years. DESIGN: Cross-sectional study. SETTING: Department of Obstetrics and Gynecology and Department of Biochemistry, Assuit university hospital. SUBJECTS: Forty-six patients had malignant ovarian tumors. Sixteen patients had benign ovarian neoplasm. Thirty patients, with normal ovaries, represented the control group. METHODS: A sample of the tumor or from the normal ovary (the control group) was sent for histopathological and biochemical examination. Sulfatides were measured by a rapid and sensitive spectrophotometric method. RESULTS: There was a significant rise in benign tumors [median and range 43 (38-53)], than in the control group, 21 (18-31), P-value = 0.000. In malignant tumors, the median value of sulfatides was significantly higher than in benign tumors [127 (71-193), P-value = 0.000]. Sulfatides were significantly higher in patients with more ovulatory years and tumors of advanced stages (stage III/IV) and poor differentiation. CONCLUSIONS: Sulfatides may play a role in the pathogenesis of benign and malignant ovarian tumors. It may also predict advanced stages in patients who are apparently early stage. It is also a candidate to study of their association with response to chemotherapy. PMID- 14764035 TI - Vulvar soft tissue tumors. AB - OBJECTIVE: To report our incidence of soft tissue tumors at this site and to discuss various prognostic factors. METHODS: All patients with a diagnosis of vulvar soft tissue neoplasms were studied from a prospective database at the Royal Marsden Hospital between January 1985 and July 2001. RESULTS: Seventeen vulvar soft tissue neoplasms (11 malignant and six benign) were treated during this period. Leiomyosarcoma (n = 5) and aggressive angiomyxoma (n = 4) were the most frequent histologic types. According to the grade, there were four G3, three G2, three G1 and could not be assessed in one patient. Local recurrence occurred in six patients with sarcoma (three with high grade and one each with intermediate, low, and undetermined grade). In this group, five patients had negative microscopic margins and one patient had positive microscopic margins on excision. All three women with low-grade sarcomas are alive without evidence of disease. Three patients with aggressive angiomyxoma also had a local recurrence. CONCLUSION: Surgical excision is the primary treatment where possible. The grade of the tumor is an important predictor for local recurrence and outcome. Aggressive angiomyxoma is a local problem. Leiomyosarcoma and aggressive angiomyxoma are the most frequent histologic types. PMID- 14764036 TI - Unique use of botulinum toxin to decrease adductor tone and allow surgical excision of vulvar carcinoma. AB - Here, we present the case of an 86-year-old woman with vulvar carcinoma requiring surgical resection and with Parkinson's disease with severe spasticity and contractures of the lower extremities. Because of the patient's severe contractures and spasticity (her knees could only be separated by 2 cm with sustained abducting force), surgical positioning and access to the vulva were impossible. The patient was admitted, intending to undergo surgery after injection with botulinum toxin (BTX) to hip adductors and intensive physical therapy. After confirmed healed hip arthroplasty, the patient underwent BTX injection (400 U) to her bilateral adductor brevis, adductor longus, adductor magnus, and semimembranosus and semitendinosus muscles on day 2 of her hospital stay. On day 3, a physical therapist began a twice-a-day stretching program. An adjustable abduction brace was custom-made to provide sustained stretching. On day 9, the patient underwent wide local excision of vulvar carcinoma with the abductor brace in place. The patient tolerated the surgery well and was discharged home on day 11 with continuous physical therapy. Upon discharge, the distance between the patient's knees was 14 cm. This unique case demonstrated a new indication for BTX treatment in the preoperative setting to allow surgical positioning and access. PMID- 14764037 TI - The risk of nodal metastasis in early adenocarcinoma of the uterine cervix. AB - A functional and widely accepted definition of microinvasive cervical adenocarcinoma remains elusive. The purpose of this study was to determine at which depth of invasion the likelihood of lymph node metastasis or disease recurrence was so small that conservative surgery could be considered appropriate. Charts of patients with adenocarcinoma of the cervix (ACC) who underwent radical hysterectomy and pelvic lymphadenectomy (n = 98) at Indiana University Medical Center from 1987 to 1998 were retrospectively reviewed. Patients with stage IA1-IB1 lesions were included in the study. Patients treated with preoperative radiotherapy were excluded. Pathologic parameters evaluated included histologic type, depth of stromal invasion (DOI), and the presence of lymphatic vascular space invasion, or lymph node metastases. The patient median age was 39 years (20-65). The median time of follow-up was 30 months (4-124). Lymph node metastases were found in ten patients and 11 developed recurrences. The precise DOI could be measured in 84 patients. Of the 48 patients with cancers with a DOI 5 mm had nodal metastases (P = 0.00069). None of these 48 patients with a tumor DOI 5 mm developed recurrent disease (P = 0.0048). The risk of nodal metastases and recurrence is so low in patients with ACC and DOI 15 years of education. The women who had p53 (Arg/Arg), p21 (Ser/Ser), and >3 children showed a 6.4-fold increased risk of cervix cancer compared to the women who had p53 ( approximately Pro), p21 ( approximately Arg), and no children. The women who had p53 (Arg/Arg), IRF-1 (T/T), and first sexual intercourse before 22 years old showed a 5.5-fold increased risk of cervix cancer compared to the women who had p53 ( approximately Pro), IRF-1 ( approximately C), and first sexual intercourse after 26 years old. CONCLUSIONS: We found that the level of education, the age at first intercourse, and the number of children were independent risk factors in cervix carcinogenesis. The specific combinations of p53, p21, and IRF-1 gene-gene and gene-environmental interactions were significantly noted in the cervix carcinogenesis of Korean women. PMID- 14764040 TI - Correlation between histological criteria and human papillomavirus presence based on PCR assay in cervical biopsies. AB - The aim was to correlate histological findings in cervix lesions to human papillomavirus (HPV), as detected by polymerase chain reaction (PCR). One hundred and seven women with atypical Pap smear were submitted to colposcopic examination, and suspicious images were biopsied. The PCR assay was performed with the primers MY09/11 and GP05/06+ and, as control, the beta-globin gene was amplified. The morphological findings were correlated to HPV positivity: parakeratosis, acanthosis, koilocytotic atypia (KA), binucleation, dyskeratosis, and number of mitoses. From 107 patients, 61 biopsies were taken: 11 chronic cervicitis (CC), 36 cervical intraepithelial neoplasia (CIN) (13 CIN I; 10 CIN II; 13 CIN III), and 14 suggestive for HPV (SHPV). DNA extraction was not possible in eight cases. HPV was found in 35% CC, 77% CIN, and 64% SHPV. The analysis did not indicate any morphological criteria strongly related to HPV. The findings with highest sensitivity for HPV were KA (88.89%) and binucleation (75%), but with low specificity of 29.41 and 52.94%, respectively. The higher predictive positive values (PV+) for HPV were also KA (72.73%) and binucleation (77.14%). Considering KA, dyskeratosis and binucleation together, PV+ was 72.41%. CONCLUSION: Although indicative, none of the studied morphological criteria was always related to PCR virus detection, denoting some limitations for histological diagnosis. PMID- 14764041 TI - Nuclear and cytoplasmic c-myc staining in endometrial carcinoma and their relationship to survival. AB - OBJECTIVE: The role of the c-myc proto-oncogene in genomic instability is just becoming more fully understood. However, its role in endometrial cancer is essentially unknown. The objective of this study was to determine the relationship between cytoplasmic and nuclear c-myc staining, DNA index, and survival in patients with endometrial carcinoma. METHODS: One hundred and twenty one patients with endometrial carcinoma were studied. Image analysis was used to determine DNA index. In addition to cytoplasmic and nuclear c-myc staining and DNA index, histologic type, stage, grade, depth of invasion, lymphvascular space invasion, and peritoneal cytology were evaluated as prognostic indicators. Univariate and multivariate analyses were performed. RESULTS: One hundred and twenty-one patients were followed for over 5 years. c-myc cytoplasmic staining was present in 75.2% of the patients' tumors, and nuclear staining was present in 66.9% (P = 0.99). DNA index was significantly higher in patients with nuclear c myc staining and no cytoplasmic staining (DNA index 1.38) as compared to those patients whose tumors displayed cytoplasmic c-myc staining but no nuclear c-myc staining (1.18) (P = 0.016). Patients whose tumors stained positively for nuclear c-myc and negatively for cytoplasmic c-myc had significantly worse survival by Kaplan-Meier analysis (P < 0.0001). Seventeen patients died during the follow-up period of this study. By multivariate analysis, positive cytoplasmic c-myc staining with negative nuclear staining (P = 0.0076), negative cytoplasmic c-myc staining with positive nuclear staining (P = 0.011) and FIGO stage (P < 0.0001) were shown to be independent prognostic indicators predictive of survival. CONCLUSION: Nuclear and cytoplasmic c-myc staining, as well as FIGO stage, when assessed by multivariate analysis, were demonstrated to be important factors in predicting survival in the 121 patients in this study. While increasing FIGO stage was prognostic of decreased survival, the specific location of c-myc staining was also associated with prognosis. The expression of the c-myc protein is related to survival in patients with adenocarcinoma of the endometrium. PMID- 14764042 TI - Nuclear size, shape, and density in endometrial carcinoma: relationship to survival at over 5 years of follow-up. Does analyzing only cells occupying the G0 G1 peak add useful information? AB - OBJECTIVE: The authors, using image analysis, previously demonstrated nuclear size and summed optical density to be independent prognostic indicators of recurrence in patients with endometrial carcinoma. The same tumors were analyzed by studying the optical features in the G0-G1 peak to see if this changed the values found as well as their importance as prognostic features at greater than 5 years of follow-up. METHODS: Tumors from 74 consecutive patients, surgically treated, with endometrial cancer, were evaluated. Survival, depth of invasion, lymphvascular space invasion, FIGO stage, grade, histology were analyzed. DNA index, progesterone receptor status, as well as nuclear size (NUSZ), shape (NUSH), and summed optical density (NUSD) were evaluated. NUSZ, NUSH, and NUSD were quantified using image analysis. RESULTS: Fifteen patients died from disease during the observation period of the study. Mean follow-up was 82 months with a median of 84 months. Forty-nine patients had stage I cancers, five stage II, 17 stage III, and three stage IV. NUSZ and NUSD were all significantly different between the original (entire cell cycle) and the re-measured (G0G1 only) values (both P < 0.001). Multivariate analysis showed both the original (P = 0.0001) and G0G1-only (P = 0.046) NUSZ and the original (P = 0.0002) and G0G1-only (P = 0.018) NUSD to be independent prognosticators of survival. CONCLUSION: Image analysis is able to quantify cellular and nuclear parameters not otherwise quantifiable. NUSD and NUSZ correlated with traditional prognostic indicators, were demonstrated independent predictors of survival at over 5 years of follow up. Although the re-measured NUSZ and NUSD from only the G0-G1 peak were significantly different from the original NUSZ and NUSD, they were not as valuable as prognostic factors. Nuclear size and summed optical density measured from the entire cell cycle are independent prognostic indicators of survival at greater than 5 years of follow-up. Measuring nuclear morphometric features in the G0-G1 peak only does not add any new prognostic information. PMID- 14764044 TI - Comparison of hepatocyte growth factor levels of epithelial ovarian cancer cyst fluids with benign ovarian cysts. AB - Hepatocyte growth factor (HGF) is known to take role in oncogenesis and tumoral behavior of the tumors of the organs that contain mesenchymal and epithelial cells together. This study aims to compare HGF levels in cyst fluids of epithelial ovarian cancer and benign ovarian cysts and look for the role of HGF in ovarian carcinogenesis. Twenty-four consecutive patients with ovarian cancer and 34 with benign cysts of ovary were recruited prospectively at Gynecologic Oncology Departments of SSK Ankara Maternity Hospital and Hacettepe University School of Medicine between 2001 and 2002. Cyst fluids were collected during primary staging in cancer patients and during laparatomy for benign patients. HGF levels were measured by enzyme-linked immunosorbent assay method. Median HGF levels of the benign ovarian cysts and epithelial ovarian tumoral fluids were found as 3822 pg/ml (85-15,253 pgr/ml) and 12,962 pgr/ml (4136-16,025 pgr/ml), respectively. Malign cyst fluids have higher HGF levels when compared with benign ovarian cysts (P < 0.01). This finding suggests that HGF may take a paracrine role in oncogenic differentiation and tumoral development of epithelial ovarian cancers. Mechanisms that take role in HGF secretion and the answers of the neighboring epithelial cells to HGF during tumoral development need to be investigated. PMID- 14764043 TI - Inability of immunohistochemistry to predict clinical outcomes of endometrial cancer patients. AB - INTRODUCTION: Despite optimal surgery, some patients with early endometrial carcinoma develop recurrence and die of disease. A number of immunohistochemical (IHC)-identified cell products (markers) have been proposed as predictors of recurrence. This study characterizes a large series of endometrial carcinomas with previously described markers as well as markers that have not been investigated in endometrial carcinoma. PATIENTS AND METHODS: Women who had undergone surgery for endometrial carcinoma were identified and specimens accessed. Tissue blocks were evaluated for ten IHC markers. Results were correlated with last known clinical status. RESULTS: Mean follow-up was 43 months; complete data were available on 117 patients. Two women died of other causes; of the remaining 115, eight died of disease and six were alive with recurrence at last follow-up (12%). Vascular endothelial growth factor staining independently predicted recurrence and death. However, in multivariate analyses, only FIGO stage predicted outcome. DISCUSSION: Our goal was to identify markers to predict which women with endometrial carcinoma were likely to have disease recurrence. We evaluated cell-cycle regulatory proteins, growth factors, hormone receptors, and angiogenic factors, but did not identify any marker that independently predicted outcome in multivariate analysis. This may reflect the few negative outcomes in our population. PMID- 14764045 TI - The role of neurosurgery in the treatment of cerebral metastases from choriocarcinoma: a report of two cases. AB - Cerebral metastases from choriocarcinoma are a poor prognostic indicator of outcome in both the World Health Organization and FIGO classification systems. However, with the increased experience with chemotherapy and radiotherapy the prognosis of this group of patients has improved substantially. Neurosurgery remains an option for selected patients. We present two patients who underwent craniotomy as part of their management of choriocarcinoma, and review the role of neurosurgery in the treatment of gestational trophoblastic disease. PMID- 14764046 TI - Malignant peritoneal mesothelioma in a 76-year-old woman with a history of asbestos fiber ingestion. AB - We report on a woman with malignant mesothelioma of the peritoneum. This is the first report of a subject with this disease who revealed a history of asbestos ingestion by asbestos-contaminated food. She presented with episodes of sweating and fever, ascites, and weight loss. At laparotomy, small tumor deposits were noted on the peritoneum. The omental cake was removed, together with the uterus, ovaries, and tubes which were all macroscopically normal. The diagnosis was established by immunohistochemistry and electron microscopy. Postoperatively, her complaints of fever and sweating disappeared. She refused further chemotherapy. After questioning her for asbestos exposure, she told us that, years ago, she used to prepare vegetables for cooking in rain water collected from a roof made of asbestos. PMID- 14764047 TI - Primary fallopian tube carcinoma presenting with a sinus in the posterior portion of cervix. AB - Primary fallopian tube carcinoma is an aggressive but rare tumor. This is an unusual presentation of papilliferous tubal carcinoma presenting with vaginal discharge through a large sinus approximately 2-3 cm in size, located posterior to the neck of cervix. Surgery was undertaken with the initial diagnosis of primary ovarian carcinoma and the fistula was left to heal spontaneously. Surgical resection was followed by adjuvant chemotherapy. The patient is doing well after the therapy. PMID- 14764048 TI - Synchronous mucinous adenocarcinoma of the endometrium and mucinous cystadenoma of bilateral ovaries presenting during fertility therapy. AB - We describe a very rare case of synchronous mucinous tumor of the endometrium and ovaries presenting during ovulation induction. A 31-year-old woman received ovulation induction for 5-year primary infertility. Ultrasonography revealed mucus retention in the uterine cavity and bilateral multicystic ovaries during ovulation induction. Atypical hyperplasia was diagnosed by endometrial curettage. Repeated procedures including ovarian cystectomy, endometrial curettage and in vitro fertilization combined with progestine therapy resulted in no pregnancy but rapid recurrences. She finally underwent simple hysterectomy and bilateral salpingo-oophorectomy. Microscopic examination revealed mucinous cystadenoma in the both ovaries and well differentiated mucinous adenocarcinoma of the endometrium. PMID- 14764049 TI - Rapid growth of myxoid leiomyosarcoma of the vulva during pregnancy: a case report. AB - Smooth muscle tumors arising in the vulva are rare. Leiomyosarcoma is the most common variant of vulvar sarcoma, and very few cases have been reported during pregnancy. A 36-year-old woman presented with a progressively enlarging vulvar mass during pregnancy, diagnosed as a Bartholin's gland cyst. The lesion was resected at 38 weeks of gestation during cesarean section and diagnosis of myxoid leiomyosarcoma of the vulva was made. Six weeks later the patients were referred to our center and submitted to wide vulvar excision with groin lymph node dissection that revealed the presence of a small residual focus of leiomyosarcoma. At 30 months of follow-up the patient was well without any sign of recurrent disease. Leiomyosarcoma should be included in the differential diagnosis of vulvar masses; progressively enlarging vulvar lesion should be biopsied even during pregnancy. Leiomyosarcoma should be considered in the differential diagnosis of vulvar mass. PMID- 14764050 TI - Extensive subcutaneous metastasis from squamous cell carcinoma of the cervix in patient with HIV. PMID- 14764051 TI - Compare 42 node-negative patients with 42 node-positive patients. PMID- 14764053 TI - Applications of microarrays to histopathology. AB - High-throughput microarray technologies have the potential to impact significantly on the practice of histopathology over the coming years. Global gene expression profiling allows for a systematic search of all human genes for novel diagnostic and prognostic markers and for potential therapeutic targets. Likewise, gene copy number changes can be determined on a gene-by-gene basis using microarrays. Tissue microarrays are an efficient method to extend and validate the findings obtained from the initial 'discovery' phase of the research, done using cDNA microarrays. In addition, tissue microarrays can be used for quality assurance for immunohistochemical and in situ hybridization procedures. In this review we give a brief overview of microarray technology and research uses, and discuss potential applications of microarrays in the practice of diagnostic histopathology. PMID- 14764054 TI - WT-1 assists in distinguishing ovarian from uterine serous carcinoma and in distinguishing between serous and endometrioid ovarian carcinoma. AB - AIMS: It has been suggested that WT-1 is helpful in distinguishing a primary ovarian serous carcinoma (OSC) from a primary uterine serous carcinoma (USC). Since both neoplasms are often disseminated at diagnosis and since USC often spreads to the ovary and vice versa, it may be difficult to ascertain the primary site. This is important, since adjuvant therapies for OSC and USC may differ. WT 1 staining patterns also differ between OSC and ovarian endometrioid carcinoma and so it is possible that WT-1 may assist in the distinction of these two neoplasms, which is sometimes problematic, especially with poorly differentiated tumours. This study aims to document the value of WT-1 in these settings. Cases of ovarian borderline serous tumour, primary peritoneal serous carcinoma (PPSC) and uterine endometrioid carcinoma were also studied. METHODS AND RESULTS: Cases of OSC (n = 38), USC (n = 25) (in five of these cases there was also a component of endometrioid adenocarcinoma), ovarian endometrioid carcinoma (n = 13), uterine endometrioid carcinoma (n = 7), ovarian borderline serous tumour (n = 16) and PPSC (n = 6) were stained with WT-1. Cases were scored on a scale of 0-3, depending on the percentage of positive cells. The intensity of staining was scored as weak, moderate or strong. There was positive nuclear staining of 36 of 38 (94.7%) OSC with WT-1. In most OSC (68.4%), >50% of cells stained positively and staining was usually strong. Five of 25 (20%) USC were positive with only two cases exhibiting staining of >50% of cells. All primary ovarian and uterine endometrioid carcinomas were negative. All PPSC were positive, usually with diffuse strong immunoreactivity. Fourteen of 16 borderline serous tumours exhibited positivity with WT-1. CONCLUSIONS: WT-1 is useful in distinguishing OSC (characteristically diffuse strong nuclear positivity) from USC (characteristically negative). However, rarely OSC is negative and occasional cases of USC are positive. WT-1 may also be helpful in differentiating poorly differentiated OSC from poorly differentiated ovarian endometrioid carcinoma. PMID- 14764055 TI - Preeclampsia is associated with loss of neuronal nitric oxide synthase expression in vascular smooth muscle cells of the human umbilical cord. AB - AIMS: Umbilical blood vessels are not innervated and regulation of blood flow to the placenta must depend on structural changes and the effect of vasoactive factors. Failure to achieve these adaptations may result in reduced fetoplacental perfusion. The purpose of this study was to determine whether neuronal nitric oxide synthase (nNOS) is expressed in human vascular smooth muscle cells (VSMCs) of the fetoplacental circulation. nNOS has been described as a non-endothelial NOS counterregulating vasoconstriction only in the VSMCs of animal models. Therefore, we investigated nNOS expression in the fetoplacental unit from preeclamptic and healthy pregnancies. METHODS AND RESULTS: We investigated nNOS regulation by immunohistochemistry, Western blotting and reverse transcriptase polymerase chain reaction analysis. nNOS activity was determined by measuring the conversion of L-3H-arginine to L-3H-citrulline. nNOS expression was revealed only in VSMCs of the human umbilical veins, but not in umbilical arteries. A more direct assessment of nNOS activity showed that a small, but consistent amount of nNOS is present in the denuded media of the umbilical vein. In VSMCs of the umbilical veins during preeclampsia a total loss of nNOS protein expression and a significant decrease in mRNA expression were seen. CONCLUSIONS: Loss of nNOS expression is associated with preeclampsia. It may alter the regulation of blood flow in the fetal and maternal placental vasculature in preeclampsia. However, the impact of NO produced by nNOS on the vascular tone of umbilical veins remains to be elucidated. PMID- 14764056 TI - Glutamate-L-cysteine ligase in breast carcinomas. AB - AIMS: To investigate the immunohistochemical expression of the catalytic and regulatory subunits of gamma-glutamyl cysteine synthetase, i.e. glutamate-L cysteine ligase (GLCL) in 274 invasive and in-situ breast carcinomas. GLCL is the rate-limiting enzyme in glutathione synthesis, which is one of the most important intracellular antioxidants participating in the detoxification reactions of several cytotoxic drugs. METHODS AND RESULTS: In the tumour cells GLCL reactivity was observed in 50% and 44% of the cases for the catalytic and the regulatory subunits, respectively. There was a statistically significant association between their expression (P = 0.002). Lobular invasive carcinomas expressed the catalytic and regulatory subunits more often than other tumours (P = 0.050 and P = 0.046, respectively). Also in-situ carcinomas expressed the catalytic subunit more often (P = 0.005). Tumours showing no immunoreactivity for the catalytic subunit had axillary metastases significantly more often (P = 0.013). Patients with tumours showing positivity for either subunit or both had a better survival (P = 0.037). No difference in survival could be observed between GCLC-positive or -negative cases in the subgroup receiving chemotherapy. CONCLUSIONS: Expression of the catalytic and regulatory subunits of GLCL is found in a substantial number of breast carcinomas and their expression is more pronounced in lobular invasive and in-situ carcinomas. Even though the overall expression of GLCL was associated with improved survival, no such effect was observed separately in the group receiving chemotherapy. PMID- 14764057 TI - Osteopontin: correlation with interstitial fibrosis in human diabetic kidney and PI3-kinase-mediated enhancement of expression by glucose in human proximal tubular epithelial cells. AB - AIMS: To examine the expression and localization of osteopontin (OPN), a secreted phosphoprotein implicated in the development of tubulointerstitial inflammation in various models of renal disease, in human diabetic kidneys, and to study the regulation of OPN expression in primary cultures of human renal proximal tubular epithelial cells (RPTEC). METHODS AND RESULTS: Differential gene expression profiling through subtractive hybridization demonstrated increased renal OPN mRNA expression in a patient with diabetic nephropathy. Immunohistochemical staining of normal and diabetic human kidney samples confirmed that OPN was localized to cortical tubular, interstitial and juxtaglomerular compartments. Quantification of OPN immunostaining revealed a marked increase in the percentage of OPN positive tubular profiles in diabetic kidneys (47 +/- 9% versus 5 +/- 3%, diabetic versus minimal change disease) that correlated strongly with the degree of cortical scarring (r2 = 0.91). Results of Northern hybridization, flow cytometry and Western blotting indicated that glucose up-regulates OPN mRNA and protein expression in primary cultures of human RPTECs. This effect was independent of the osmotic effects of glucose and independent of insulin. Finally, glucose-stimulated OPN expression was inhibited by LY294002, an inhibitor of phosphatidylinositol 3-kinase activity, in a dose-dependent manner. CONCLUSIONS: OPN is expressed in human diabetic kidneys and regulation of OPN expression is via a glucose-mediated, phosphatidylinositol 3-kinase-dependent pathway. PMID- 14764058 TI - Combined status of MUC1 mucin and surfactant apoprotein A expression can predict the outcome of patients with small-size lung adenocarcinoma. AB - AIM: Lung cancer is still a disease of high mortality, despite advanced diagnostic techniques. Here, we aim to report a unique method to predict the recurrence and outcome of patients with pulmonary adenocarcinomas. METHODS AND RESULTS: Immunohistochemical expression of MUC1 mucin and surfactant apoprotein A (SP-A) was examined in 185 cases of surgically removed lung adenocarcinomas of non-bronchioloalveolar type smaller than 30 mm. Staining results were evaluated semiquantitatively, and the expression of MUC1 and SP-A was compared in each case. There were 140/185 (76%) cases showing MUC1 expression higher than SP-A expression (MUC1>SP-A), and 45/185 (24%) cases showing MUC1 expression lower than or equal to SP-A expression (MUC1SP-A pattern, but in 7% (3/45) of the patients with a MUC1< or =SP-A pattern after the median observation period of 41 months (1-99 months) (P < 0.01). The MUC1>SP-A group showed higher recurrence and worse survival than the MUC1 0.5). However, cerebral oxygen saturation increased from 40.4% to 49.6% (P = 0.01) and correlated significantly with the haemoglobin level (r = 0.553, P = 0.003) in 14 subjects studied before and after transfusions. In seven subjects who received simple transfusions, cerebral oxygen saturation correlated strongly and positively with the haemoglobin level (r = 0.811, P = 0.001) and with percent normal haemoglobin (r = 0.786, P = 0.002), and negatively with abnormal sickle haemoglobin (r = -0.775, P = 0.003). None of these correlations was found to be statistically significant in the seven subjects given exchange transfusions. Cerebral oxygen saturation measured in the sickle cell disease subjects after transfusions was still significantly lower than in the anaemic subjects without sickle cell disease and in the normal controls (49.6% vs. 59.8% and 61.3%, P = 0.001). CONCLUSIONS: We found that patients with sickle cell disease have subnormal values of cerebral oxygen saturation. Red cell transfusions significantly increased the brain oxygenation in these patients. Cerebral oximetry may be a useful, noninvasive method for assessing the effect of circulating normal red cells in sickle cell patients after transfusions. PMID- 14764079 TI - Genetic deficiency of CD16, the low-affinity receptor for immunoglobulin G, has no impact on the functional capacity of polymorphonuclear neutrophils. AB - BACKGROUND: Of the three receptors for immunoglobulin G (IgG), the low-affinity receptor CD16 is constitutively expressed on polymorphonuclear neutrophils (PMNs), monocytes and NK-cells. CD16 participates in various effector functions, notably phagocytosis of opsonized particles or of immune complexes, and in antibody-dependent cellular cytotoxicity (ADCC). In the present study we report a case of total CD16 deficiency on PMNs and monocytes. DESIGN: Polymorphonuclear neutrophils, monocytes and NK-cells were analyzed for surface-receptor expression by cytofluorometry and laser scan microscopy. Moreover, CD16-specific mRNA was assessed by RT-PCR. As functional parameters, phagocytosis of opsonized bacteria was tested, as was superoxide production. RESULTS: Polymorphonuclear neutrophils and monocytes totally deficient in CD16 were detected by chance in an apparently healthy individual. Further analysis revealed that two more members of his family, his father and sister, were also deficient in CD16. All were healthy and there was no evidence of an increased frequency, or of exceptionally severe or persistent infections. Despite the lack of CD16, phagocytosis of antibody-coated bacteria was within the normal range, as was the superoxide production. CONCLUSION: Deficiency of CD16 does not compromise the host defence. Apparently, the other receptors for IgG, CD32 and CD64, can compensate for the lack of CD16. PMID- 14764080 TI - High levels of desmosines in urine and plasma of patients with pseudoxanthoma elasticum. AB - BACKGROUND: Pseudoxanthoma elasticum (PXE), a rare heritable disorder caused by mutations of the ABCC6 gene, is characterized by fragmentation and mineralization of elastic fibres. We determined the extent of degradation of elastin by measuring and comparing the amount of desmosines in plasma and urine of PXE patients, healthy carriers and normal subjects. METHODS: Using capillary electrophoresis with laser-induced fluorescence detection (CE-LIF) we measured the amount of desmosines in the urine of 46 individuals (14 PXE patients, 17 healthy carriers and 15 controls) and in the plasma of 56 subjects (18 PXE patients, 23 healthy carriers and 15 controls). Pseudoxanthoma elasticum patients and carriers were identified by clinical, structural and molecular biology analyses. RESULTS: The urinary excretion of desmosines was two-fold higher in PXE patients than in controls (P < 0.01); the values for healthy carriers were intermediate between those of PXE patients and controls. A very similar trend between patients and their relatives was observed for plasma desmosines. There was a significant correlation between the amount of the desmosines in plasma and urine. Moreover, a positive correlation was observed between urinary desmosine content and age of the patients as well as between urinary desmosine content and severity of clinical manifestations. CONCLUSIONS: Both the urinary and plasma desmosine concentrations indicate that elastin degradation is higher in PXE patients and, to a lesser extent, in healthy carriers than in normal subjects. Data seem to indicate that the amount of elastin breakdown products correlates with the age of patients as well as with the severity of the disease. PMID- 14764081 TI - Cessation of passive smoking reduces allergic responses and plasma neurotrophin. PMID- 14764082 TI - Expressed protein ligation. Method and applications. AB - The introduction of noncanonical amino acids and biophysical probes into peptides and proteins, and total or segmental isotopic labelling has the potential to greatly aid the determination of protein structure, function and protein-protein interactions. To obtain a peptide as large as possible by solid-phase peptide synthesis, native chemical ligation was introduced to enable synthesis of proteins of up to 120 amino acids in length. After the discovery of inteins, with their self-splicing properties and their application in protein synthesis, the semisynthetic methodology, expressed protein ligation, was developed to circumvent size limitation problems. Today, diverse expression vectors are available that allow the production of N- and C-terminal fragments that are needed for ligation to produce large amounts and high purity protein(s) (protein alpha-thioesters and peptides or proteins with N-terminal Cys). Unfortunately, expressed protein ligation is still limited mainly by the requirement of a Cys residue. Of course, additional Cys residues can be introduced into the sequence by site directed mutagenesis or synthesis, however, those mutations may disturb protein structure and function. Recently, alternative ligation approaches have been developed that do not require Cys residues. Accordingly, it is theoretically possible to obtain each modified protein using ligation strategies. PMID- 14764083 TI - The role of N-linked glycosylation in the protection of human and bovine lactoferrin against tryptic proteolysis. AB - Lactoferrin (LF) is an iron-binding glycoprotein of the innate host defence system. To elucidate the role of N-linked glycosylation in protection of LF against proteolysis, we compared the tryptic susceptibility of human LF (hLF) variants from human milk, expressed in human 293(S) cells or in the milk of transgenic mice and cows. The analysis revealed that recombinant hLF (rhLF) with mutations Ile130-->Thr and Gly404-->Cys was about twofold more susceptible than glycosylated and unglycosylated variants with the naturally occurring Ile130 and Gly404. Hence, N-linked glycosylation is not involved in protection of hLF against tryptic proteolysis. Apparently, the previously reported protection by N linked glycosylation of hLF [van Berkel, P.H.C., Geerts, M.E.J., van Veen, H.A., Kooiman, P.M., Pieper, F., de Boer, H.A. & Nuijens, J.H. (1995) Biochem. J. 312, 107-114] is restricted to rhLF containing the Thr130 and Cys404. Comparison of the tryptic proteolysis of hLF and bovine LF (bLF) revealed that hLF is about 100 fold more resistant than bLF. Glycosylation variants A and B of bLF differed by about 10-fold in susceptibility to trypsin. This difference is due to glycosylation at Asn281 in bLF-A. Hence, glycosylation at Asn281 protects bLF against cleavage by trypsin at Lys282. PMID- 14764084 TI - Differing involvement of sulfoquinovosyl diacylglycerol in photosystem II in two species of unicellular cyanobacteria. AB - Sulfoquinovosyl diacylglycerol (SQDG) is involved in the maintenance of photosystem II (PSII) activity in Chlamydomonas reinhardtii[Minoda, A., Sato, N., Nozaki, H., Okada, K., Takahashi, H., Sonoike, K. & Tsuzuki, M. et al. (2002) Eur. J. Biochem.269, 2353-2358]. To understand the spread of the taxa in which PSII interacts with SQDG, especially in cyanobacteria, we produced a mutant defective in the putative sqdB gene responsible for SQDG synthesis from two cyanobacteria, Synechocystis sp. PCC6803 and Synechococcus sp. PCC7942. The mutant of PCC6803, designated SD1, lacked SQDG synthetic ability and required SQDG supplementation for its growth. After transfer from SQDG-supplemented to SQDG-free conditions, SD1 showed decreased net photosynthetic and PSII activities on a chlorophyll (Chl) basis with a decrease in the SQDG content. Moreover, the sensitivity of PSII activity to 3-(3,4-dichlorophenyl)-1,1-dimethylurea and atrazine was increased in SD1. However, SD1 maintained normal amounts of cytochrome b559 and D1 protein (the subunits comprising the PSII complex) on a Chl basis, indicating that the PSII complex content changed little, irrespective of a decrease in the SQDG content. These results suggest that the role of SQDG is the conservation of the PSII properties in PCC6803, consistent with the results obtained with C. reinhardtii. In contrast, the SQDG-null mutant of PCC7942 showed the normal level of PSII activity with little effect on its sensitivity to PSII herbicides. Therefore, the difference in the SQDG requirement for PSII is species specific in cyanobacteria; this could be of use when investigating the molecular evolution of the PSII complex. PMID- 14764085 TI - The unusual methanogenic seryl-tRNA synthetase recognizes tRNASer species from all three kingdoms of life. AB - The methanogenic archaea Methanococcus jannaschii and M. maripaludis contain an atypical seryl-tRNA synthetase (SerRS), which recognizes eukaryotic and bacterial tRNAsSer, in addition to the homologous tRNASer and tRNASec species. The relative flexibility in tRNA recognition displayed by methanogenic SerRSs, shown by aminoacylation and gel mobility shift assays, indicates the conservation of some serine determinants in all three domains. The complex of M. maripaludis SerRS with the homologues tRNASer was isolated by gel filtration chromatography. Complex formation strongly depends on the conformation of tRNA. Therefore, the renaturation conditions for in vitro transcribed tRNASer(GCU) isoacceptor were studied carefully. This tRNA, unlike many other tRNAs, is prone to dimerization, possibly due to several stretches of complementary oligonucleotides within its sequence. Dimerization is facilitated by increased tRNA concentration and can be diminished by fast renaturation in the presence of 5 mm magnesium chloride. PMID- 14764086 TI - Cloning and characterization of two distinct isoforms of rainbow trout heat shock factor 1. Evidence for heterotrimer formation. AB - To elucidate the molecular mechanism underlying the heat shock response in cold water fish species, genes encoding heat shock transcription factors (HSFs) were cloned from RTG-2 cells of the rainbow trout Oncorhynchus mykiss. Consequently, two distinct HSF1 genes, named HSF1a and HSF1b, were identified. The predicted amino acid sequence of HSF1a shows 86.4% identity to that of HSF1b. The two proteins contained the general structural motifs of HSF1, i.e. a DNA-binding domain, hydrophobic heptad repeats and nuclear localization signals. Southern blot analysis showed that each HSF1 is encoded by a distinct gene. The two HSF1 mRNAs were coexpressed in unstressed rainbow trout RTG-2 cells and in various tissues. In an electrophoretic mobility shift assay, each in vitro translated HSF1 bound to the heat shock element. Chemical cross-linking and immunoprecipitation analysis showed that HSF1a and HSF1b form heterotrimers as well as homotrimers. Taken together, these results demonstrate that in rainbow trout cells there are two distinct HSF1 isoforms that can form heterotrimers, suggesting that a unique molecular mechanism underlies the stress response in tetraploid and/or cold-water fish species. PMID- 14764087 TI - Non-specific depolymerization of chitosan by pronase and characterization of the resultant products. AB - Pronase (type XXV serine protease from Streptomyces griseus) efficiently depolymerizes chitosan, a linear beta-->1,4-linked polysaccharide of 2-amino deoxyglucose and 2-amino-2-N-acetylamino-D-glucose, to low-molecular weight chitosans (LMWC), chito-oligomers (degree of polymerization, 2-6) and monomer. The maximum depolymerization occurred at pH 3.5 and 37 degrees C, and the reaction obeyed Michaelis-Menten kinetics with a Km of 5.21 mg.mL(-1) and Vmax of 138.55 nmoles.min(-1).mg(-1). The molecular mass of the major product, LMWC, varied between 9.0 +/- 0.5 kDa depending on the reaction time. Scanning electron microscopy of LMWC showed an approximately eightfold decrease in particle size and characterization by infrared spectroscopy, circular dichroism, X-ray diffractometry and 13C-NMR revealed them to possess a lower degree of acetylation, hydration and crystallinity compared to chitosan. Chitosanolysis by pronase is an alternative and inexpensive method to produce a variety of chitosan degradation products that have wide and varied biofunctionalities. PMID- 14764088 TI - The zinc-binding site of a class I aminoacyl-tRNA synthetase is a SWIM domain that modulates amino acid binding via the tRNA acceptor arm. AB - In its tRNA acceptor end binding domain, the glutamyl-tRNA synthetase (GluRS) of Escherichia coli contains one atom of zinc that holds the extremities of a segment (Cys98-x-Cys100-x24-Cys125-x-His127) homologous to the Escherichia coli glutaminyl-tRNA synthetase (GlnRS) loop where a leucine residue stabilizes the peeled-back conformation of tRNAGln acceptor end. We report here that the GluRS zinc-binding region belongs to the novel SWIM domain family characterized by the signature C-x-C-xn-C-x-H (n = 6-25), and predicted to interact with DNA or proteins. In the presence of tRNAGlu, the GluRS C100Y variant has a lower affinity for l-glutamate than the wild-type enzyme, with Km and Kd values increased 12- and 20-fold, respectively. On the other hand, in the absence of tRNAGlu, glutamate binds with the same affinity to the C100Y variant and to wild type GluRS. In the context of the close structural and mechanistic similarities between GluRS and GlnRS, these results indicate that the GluRS SWIM domain modulates glutamate binding to the active site via its interaction with the tRNAGlu acceptor arm. Phylogenetic analyses indicate that ancestral GluRSs had a strong zinc-binding site in their SWIM domain. Considering that all GluRSs require a cognate tRNA to activate glutamate, and that some of them have different or no putative zinc-binding residues in the corresponding positions, the properties of the C100Y variant suggest that the GluRS SWIM domains evolved to position correctly the tRNA acceptor end in the active site, thereby contributing to the formation of the glutamate binding site. PMID- 14764089 TI - Conformational changes of beta-lactoglobulin in sodium bis(2-ethylhexyl) sulfosuccinate reverse micelles. A fluorescence and CD study. AB - The effect of beta-lactoglobulin encapsulation in sodium bis(2-ethylhexyl) sulfosuccinate reverse micelles on the environment of protein and on Trp was analysed at different water contents (omega0). CD data underlined the distortion of the beta-sheet and a less constrained tertiary structure as the omega0 increased, in agreement with a concomitant red shift and a decrease in the signal intensity obtained in steady-state fluorescence measurements. Fluorescence lifetimes, evaluated by biexponential analysis, were tau1 = 1.28 ns and tau2 = 3.36 ns in neutral water. In reverse micelles, decay-associated spectra indicated the occurrence of important environmental changes associated with omega0. Bimolecular fluorescence quenching by CCl4 and acrylamide was employed to analyse alterations in the accessibility of the two Trp residues in beta-lactoglobulin, induced by changes in omega0. The average bimolecular quenching constant was found not to depend on omega0, confirming the insolubility of this quencher in the aqueous interface, while increases with omega0. The drastic decrease with omega0 of kq, associated with the longest lifetime kq2(CCl4), comparatively to the increase of kq2(acrylamide), emphasizes the location of beta-lactoglobulin in the aqueous interfacial region especially at omega0> or = 10. The fact that (omega0 = 30) >> kq2(acrylamide) (water) also confirms the important conformational changes of encapsulated beta-lactoglobulin. PMID- 14764090 TI - Characterization of the products of the genes SNO1 and SNZ1 involved in pyridoxine synthesis in Saccharomyces cerevisiae. AB - Genes SNO1 and SNZ1 are Saccharomyces cerevisiae homologues of PDX2 and PDX1 which participate in pyridoxine synthesis in the fungus Cercospora nicotianae. In order to clarify their function, the two genes SNO1 and SNZ1 were expressed in Escherichia coli either individually or simultaneously and with or without a His tag. When expressed simultaneously, the two protein products formed a complex and showed glutaminase activity. When purified to homogeneity, the complex exhibited a specific activity of 480 nmol.mg(-1).min(-1) as glutaminase, with a Km of 3.4 mm for glutamine. These values are comparable to those for other glutamine amidotransferases. In addition, the glutaminase activity was impaired by 6-diazo 5-oxo-L-norleucine in a time- and dose-dependent manner and the enzyme was protected from deactivation by glutamine. These data suggest strongly that the complex of Sno1p and Snz1p is a glutamine amidotransferase with the former serving as the glutaminase, although the activity was barely detectable with Sno1p alone. The function of Snz1p and the amido acceptor for ammonia remain to be identified. PMID- 14764091 TI - UDPgalactose 4-epimerase from Saccharomyces cerevisiae. A bifunctional enzyme with aldose 1-epimerase activity. AB - UDPgalactose 4-epimerase (epimerase) catalyzes the reversible conversion between UDPgalactose and UDPglucose and is an important enzyme of the galactose metabolic pathway. The Saccharomyces cerevisiae epimerase encoded by the GAL10 gene is about twice the size of either the bacterial or human protein. Sequence analysis indicates that the yeast epimerase has an N-terminal domain (residues 1-377) that shows significant similarity with Escherichia coli and human UDPgalactose 4 epimerase, and a C-terminal domain (residues 378-699), which shows extensive identity to either the bacterial or human aldose 1-epimerase (mutarotase). The S. cerevisiae epimerase was purified to > 95% homogeneity by sequential chromatography on DEAE-Sephacel and Resource-Q columns. Purified epimerase preparations showed mutarotase activity and could convert either alpha-d-glucose or alpha-d-galactose to their beta-anomers. Induction of cells with galactose led to simultaneous enhancement of both epimerase and mutarotase activities. Size exclusion chromatography experiments confirmed that the mutarotase activity is an intrinsic property of the yeast epimerase and not due to a copurifying endogenous mutarotase. When the purified protein was treated with 5'-UMP and l-arabinose, epimerase activity was completely lost but the mutarotase activity remained unaffected. These results demonstrate that the S. cerevisiae UDPgalactose 4 epimerase is a bifunctional enzyme with aldose 1-epimerase activity. The active sites for these two enzymatic activities are located in different regions of the epimerase holoenzyme. PMID- 14764092 TI - Mouse spermine oxidase gene splice variants. Nuclear subcellular localization of a novel active isoform. AB - Spermine oxidase (SMO) is a flavoenzyme involved in polyamine homeostasis in animal cells. The mouse spermine oxidase gene (mSMO) codes for splice variants, including the previously reported major active isoform, herein named alfa (alpha). In the present work, eight additional gene splicing variants were characterized. The heterologous expression and biochemical characterization of three recombinant isoforms (namely mSMOmu, -gamma and -delta) revealed that only the recombinant protein mSMO micro displays biochemical characteristics similar to those of mSMOalpha; the other two recombinant proteins contained no detectable SMO activity. In order to investigate in greater detail, the SMO enzyme activity associated with their subcellular localization, mSMOalpha and -mu V5-tagged proteins were transiently and stably transfected in the murine neuroblastoma cell line, N18TG2. Very interestingly, the novel active mSMOmu isoform was found to be present in both nuclear and cytoplasmic compartments, thus providing the first evidence of SMO activity in the nucleus, while a cytoplasmic localization was confirmed for the mSMOalpha isoform. In addition, the relative transcription levels of the gene splicing variants were evaluated by RT-PCR analysis to verify a relationship with the SMO enzyme activity in various murine organs. PMID- 14764093 TI - Identification of residues controlling transport through the yeast aquaglyceroporin Fps1 using a genetic screen. AB - Aquaporins and aquaglyceroporins mediate the transport of water and solutes across biological membranes. Saccharomyces cerevisiae Fps1 is an aquaglyceroporin that mediates controlled glycerol export during osmoregulation. The transport function of Fps1 is rapidly regulated by osmotic changes in an apparently unique way and distinct regions within the long N- and C-terminal extensions are needed for this regulation. In order to learn more about the mechanisms that control Fps1 we have set up a genetic screen for hyperactive Fps1 and isolated mutations in 14 distinct residues, all facing the inside of the cell. Five of the residues lie within the previously characterized N-terminal regulatory domain and two mutations are located within the approach to the first transmembrane domain. Three mutations cause truncation of the C-terminus, confirming previous studies on the importance of this region for channel control. Furthermore, the novel mutations identify two conserved residues in the channel-forming B-loop as critical for channel control. Structural modelling-based rationalization of the observed mutations supports the notion that the N-terminal regulatory domain and the B-loop could interact in channel control. Our findings provide a framework for further genetic and structural analysis to better understand the mechanism that controls Fps1 function by osmotic changes. PMID- 14764094 TI - A new UV-B absorbing mycosporine with photo protective activity from the lichenized ascomycete Collema cristatum. AB - A novel photo protective mycosporine was isolated from the lichenized ascomycete Collema cristatum. Biological activity was measured in terms of protection against UV-B induced membrane destruction and pyrimidine dimer formation in cultured human keratinocytes, and prevention of UV-B induced erythema. It was found that the pure isolated compound prevented UV-B induced cell destruction in a dose-dependent manner, that the compound partially prevented pyrimidine dimer formation and completely prevented UV-B induced erythema when applied to the skin prior to irradiation. PMID- 14764095 TI - ICAM-1 expression is highly NF-kappaB-dependent in A549 cells. No role for ERK and p38 MAPK. AB - The transcription factor nuclear factor kappaB (NF-kappaB) is an activator of multiple cytokines, chemokines and adhesion molecules, which are important in inflammatory diseases such as asthma, and is consequently considered as an attractive therapeutic target. In the present study, a constitutively active dominant version of IkappaBalpha, IkappaBalphaDN, was introduced into A549 pulmonary cells by adenovirus-mediated delivery. The dominant IkappaB, but not a null viral vector, prevented the induction of NF-kappaB-dependent transcription by both tumor necrosis factor alpha (TNFalpha) and interleukin-1beta (IL-1beta). Similarly, both TNFalpha and IL-1beta strongly induced mRNA and protein expression of intercellular adhesion molecule (ICAM)-1 and in each case this was prevented by adenovirus expressing the dominant IkappaB, but not by the null virus, thereby establishing ICAM-1 as an NF-kappaB-dependent gene. Numerous studies have suggested key roles for the p38 and extracellular regulated kinase (ERK) mitogen-activated protein kinase (MAPK) cascades in the activation and transactivation of NF-kappaB. We show here that SB203580, a selective inhibitor of the p38 MAPK, and PD098059 and UO126, both selective inhibitors of the ERK MAPK cascade, have no effect on TNFalpha or IL-1beta-induced translocation and DNA binding of NF-kappaB. Furthermore, these inhibitors showed no pharmacologically relevant effect on NF-kappaB-dependent transcription nor was there any effect on expression of ICAM-1. Taken together these data highlight the potential use of inhibition of the NF-kappaB signalling pathway in pulmonary inflammatory diseases and suggest that inhibitors of the p38 and ERK MAPK pathways may be of lesser effect. PMID- 14764096 TI - FRET evidence for a conformational change in TFIIB upon TBP-DNA binding. AB - As a critical step of the preinitiation complex assembly in transcription, the general transcription factor TFIIB forms a complex with the TATA-box binding protein (TBP) bound to a promoter element. Transcriptional activators such as the herpes simplex virus VP16 facilitate this complex formation through conformational activation of TFIIB, a focal molecule of transcriptional initiation and activation. Here, we used fluorescence resonance energy transfer to investigate conformational states of human TFIIB fused to enhanced cyan fluorescent protein and enhanced yellow fluorescent protein at its N- and C terminus, respectively. A significant reduction in fluorescence resonance energy transfer ratio was observed when this fusion protein, hereafter named CYIIB, was mixed with promoter-loaded TBP. The rate for the TFIIB-TBP-DNA complex formation is accelerated drastically by GAL4-VP16 and is also dependent on the type of promoter sequences. These results provide compelling evidence for a 'closed-to open' conformational change of TFIIB upon binding to the TBP-DNA complex, which probably involves alternation of the spatial orientation between the N-terminal zinc ribbon domain and the C-terminal conserved core domain responsible for direct interactions with TBP and a DNA element. PMID- 14764097 TI - Selective release and function of one of the two FMN groups in the cytoplasmic NAD+-reducing [NiFe]-hydrogenase from Ralstonia eutropha. AB - The soluble, cytoplasmic NAD+-reducing [NiFe]-hydrogenase from Ralstonia eutropha is a heterotetrameric enzyme (HoxFUYH) and contains two FMN groups. The purified oxidized enzyme is inactive in the H2-NAD+ reaction, but can be activated by catalytic amounts of NADH. It was discovered that one of the FMN groups (FMN-a) is selectively released upon prolonged reduction of the enzyme with NADH. During this process, the enzyme maintained its tetrameric form, with one FMN group (FMN b) firmly bound, but it lost its physiological activity--the reduction of NAD+ by H2. This activity could be reconstituted by the addition of excess FMN to the reduced enzyme. The rate of reduction of benzyl viologen by H2 was not dependent on the presence of FMN-a. Enzyme devoid of FMN-a could not be activated by NADH. As NADH-dehydrogenase activity was not dependent on the presence of FMN-a, and because FMN-b did not dissociate from the reduced enzyme, we conclude that FMN-b is functional in the NADH-dehydrogenase activity catalyzed by the HoxFU dimer. The possible function of FMN-a as a hydride acceptor in the hydrogenase reaction catalyzed by the HoxHY dimer is discussed. PMID- 14764098 TI - Nonlysine-analog plasminogen modulators promote autoproteolytic generation of plasmin(ogen) fragments with angiostatin-like activity. AB - We recently discovered several nonlysine-analog conformational modulators for plasminogen. These include SMTP-6, thioplabin B and complestatin that are low molecular mass compounds of microbial origin. Unlike lysine-analog modulators, which increase plasminogen activation but inhibit its binding to fibrin, the nonlysine-analog modulators enhance both activation and fibrin binding of plasminogen. Here we show that some nonlysine-analog modulators promote autoproteolytic generation of plasmin(ogen) derivatives with its catalytic domain undergoing extensive fragmentation (PMDs), which have angiostatin-like anti endothelial activity. The enhancement of urokinase-catalyzed plasminogen activation by SMTP-6 was followed by rapid inactivation of plasmin due to its degradation mainly in the catalytic domain, yielding PMD with a molecular mass ranging from 68 to 77 kDa. PMD generation was observed when plasmin alone was treated with SMTP-6 and was inhibited by the plasmin inhibitor aprotinin, indicating an autoproteolytic mechanism in PMD generation. Thioplabin B and complestatin, two other nonlysine-analog modulators, were also active in producing similar PMDs, whereas the lysine analog 6-aminohexanoic acid was inactive while it enhanced plasminogen activation. Peptide sequencing and mass spectrometric analyses suggested that plasmin fragmentation was due to cleavage at Lys615-Val616, Lys651-Leu652, Lys661-Val662, Lys698-Glu699, Lys708-Val709 and several other sites mostly in the catalytic domain. PMD was inhibitory to proliferation, migration and tube formation of endothelial cells at concentrations of 0.3-10 microg.mL(-1). These results suggest a possible application of nonlysine-analog modulators in the treatment of cancer through the enhancement of endogenous plasmin(ogen) fragment formation. PMID- 14764100 TI - Identification of a gene encoding Lon protease from Brevibacillus thermoruber WR 249 and biochemical characterization of its thermostable recombinant enzyme. AB - A gene encoding thermostable Lon protease from Brevibacillus thermoruber WR-249 was cloned and characterized. The Br. thermoruber Lon gene (Bt-lon) encodes an 88 kDa protein characterized by an N-terminal domain, a central ATPase domain which includes an SSD (sensor- and substrate-discrimination) domain, and a C-terminal protease domain. The Bt-lon is a heat-inducible gene and may be controlled under a putative Bacillus subtilis sigmaA-dependent promoter, but in the absence of CIRCE (controlling inverted repeat of chaperone expression). Bt-lon was expressed in Escherichia coli, and its protein product was purified. The native recombinant Br. thermoruber Lon protease (Bt-Lon) displayed a hexameric structure. The optimal temperature of ATPase activity for Bt-Lon was 70 degrees C, and the optimal temperature of peptidase and DNA-binding activities was 50 degrees C. This implies that the functions of Lon protease in thermophilic bacteria may be switched, depending on temperature, to regulate their physiological needs. The peptidase activity of Bt-Lon increases substantially in the presence of ATP. Furthermore, the substrate specificity of Bt-Lon is different from that of E. coli Lon in using fluorogenic peptides as substrates. Notably, the Bt-Lon protein shows chaperone-like activity by preventing aggregation of denatured insulin B chain in a dose-dependent and ATP-independent manner. In thermal denaturation experiments, Bt-Lon was found to display an indicator of thermostability value, Tm of 71.5 degrees C. Sequence comparison with mesophilic Lon proteases shows differences in the rigidity, electrostatic interactions, and hydrogen bonding of Bt-Lon relevant to thermostability. PMID- 14764099 TI - Insight into the activation mechanism of Bordetella pertussis adenylate cyclase by calmodulin using fluorescence spectroscopy. AB - The interaction of the adenylate cyclase catalytic domain (AC) of the Bordetella pertussis major exotoxin with its activator calmodulin (CaM) was studied by time resolved fluorescence spectroscopy using three fluorescent groups located in different regions of AC: tryptophan residues (W69 and W242), a nucleotide analogue (3'-anthraniloyl-2'-deoxyadenosine 5'-triphosphate, Ant-dATP) and a cysteine-specific probe (acrylodan). CaM binding elicited large changes in the dynamics of W242, which dominates the fluorescence emission of both AC and AC CaM, similar to that observed for isolated CaM-binding sequences of different lengths [Bouhss, A., Vincent, M., Munier, H., Gilles, A.M., Takahashi, M., Barzu, O., Danchin, A. & Gallay, J. (1996) Eur. J. Biochem.237, 619-628]. In contrast, Ant-dATP remains completely immobile and inaccessible to the solvent in both the AC and AC-CaM nucleotide-binding sites. As AC contains no cysteine residue, a single-Cys mutant at position 75 was constructed which allowed labeling of the catalytic domain with acrylodan. Its environment is strongly apolar and rigid, and only slightly affected by CaM. The protein's hydrodynamic properties were also studied by fluorescence anisotropy decay measurements. The average Brownian rotational correlation times of AC differed significantly according to the probe used (19 ns for W242, 25 ns for Ant-dATP, and 35 ns for acrylodan), suggesting an elongated protein shape (axial ratio of approximately 1.9). These values increased greatly with the addition of CaM (39 ns for W242, 60-70 ns for Ant-dATP and 56 ns for acrylodan). This suggests that (a) the orientation of the probes is altered with respect to the protein axes and (b) the protein becomes more elongated with an axial ratio of approximately 2.4. For comparison, the hydrodynamic properties of the anthrax AC exotoxin were computed by a mathematical approach (hydropro), which uses the 3D structure [Drum, C.L., Yan, S.-Z., Bard, J., Shen, Y.-Q., Lu, D., Soelalman, S., Grabarek, Z., Bohm, A. & Tang, W.-J. (2002) Nature (London)415, 396-402]. A change in axial ratio is also observed on CaM binding, but in the reverse direction from that for AC: from 1.7 to 1.3. The mechanisms of activation of the two proteins by CaM may therefore be different. PMID- 14764101 TI - Acharan sulfate, the new glycosaminoglycan from Achatina fulica Bowdich 1822. Structural heterogeneity, metabolic labeling and localization in the body, mucus and the organic shell matrix. AB - Acharan sulfate, a recently discovered glycosaminoglycan isolated from Achatina fulica, has a major disaccharide repeating unit of -->4)-2-acetyl,2-deoxy-alpha-d glucopyranose(1-->4)-2-sulfo-alpha-l-idopyranosyluronic acid (1-->, making it structurally related to both heparin and heparan sulfate. It has been suggested that this glycosaminoglycan is polydisperse, with an average molecular mass of 29 kDa and known minor disaccharide sequence variants containing unsulfated iduronic acid. Acharan sulfate was found to be located in the body of this species using alcian blue staining and it was suggested to be the main constituent of the mucus. In the present work, we provide further information on the structure and compartmental distribution of acharan sulfate in the snail body. Different populations of acharan sulfate presenting charge and/or molecular mass heterogeneities were isolated from the whole body, as well as from mucus and from the organic shell matrix. A minor glycosaminoglycan fraction susceptible to degradation by nitrous acid was also purified from the snail body, suggesting the presence of N-sulfated glycosaminoglycan molecules. In addition, we demonstrate the in vivo metabolic labeling of acharan sulfate in the snail body after a meal supplemented with [35S]free sulfate. This simple approach might be applied to the study of acharan sulfate biosynthesis. Finally, we developed histochemical assays to localize acharan sulfate in the snail body by metachromatic staining and by histoautoradiography following metabolic radiolabeling with [35S]sulfate. Our results show that acharan sulfate is widely distributed among several organs. PMID- 14764102 TI - Effect of synthetic peptides corresponding to residues 313-332 of the alphaIIb subunit on platelet activation and fibrinogen binding to alphaIIbbeta3. AB - The platelet integrin receptor alphaIIbbeta3 plays a critical role in thrombosis and haemostasis by mediating interactions between platelets and several ligands but primarily fibrinogen. It has been shown previously that the YMESRADR KLAEVGRVYLFL (313-332) sequence of the alphaIIb subunit plays an important role in platelet activation, fibrinogen binding and alphaIIbbeta3-mediated outside-in signalling. Furthermore, we recently showed that the 20-residue peptide (20-mer) alphaIIb 313-332, is a potent inhibitor of platelet aggregation and fibrinogen binding to alphaIIbbeta3, interacting with fibrinogen rather than the receptor. In an effort to determine the sequence and the minimum length required for the biological activity of the above 20-mer, we synthesized seven octapeptides, each overlapping by six residues, covering the entire sequence and studied their effect on platelet activation as well as fibrinogen binding to activated platelets. We show for the first time that octapeptides containing the RAD sequence are capable of inhibiting platelet aggregation and secretion as well as fibrinogen binding to the activated alphaIIbbeta3, possibly interacting with the ligand rather than the receptor. This suggests that the RAD sequence, common to all the inhibitory peptides, is critical for their biological activity. However, the presence of the YMES sequence, adjacent to RAD, significantly increases the peptide's biological potency. The development of such inhibitors derived from the 313-332 region of the alphaIIb subunit may be advantageous against the RGD-like antagonists as they could inhibit platelet activation without interacting with alphaIIbbeta3, thus failing to further induce alphaIIbbeta3-mediated outside-in signalling. PMID- 14764103 TI - Vertical-scanning mutagenesis of amino acids in a model N-myristoylation motif reveals the major amino-terminal sequence requirements for protein N myristoylation. AB - In order to determine the amino-terminal sequence requirements for protein N myristoylation, site-directed mutagenesis of the N-terminal region was performed using tumor necrosis factor (TNF) mutants as model substrate proteins. Subsequently, the susceptibility of these mutants to protein N-myristoylation was evaluated by metabolic labeling in an in vitro translation system using rabbit reticulocyte lysate. A TNF mutant having the sequence MGAAAAAAAA at its N terminus was used as the starting sequence to identify elements critical for protein N-myristoylation. Sequential vertical-scanning mutagenesis of amino acids at a distinct position in this model N-terminal sequence revealed the major sequence requirements for protein N-myristoylation: the combination of amino acids at position 3 and 6 constitutes a major determinant for the susceptibility to protein N-myristoylation. When Ser was located at position 6, 11 amino acids (Gly, Ala, Ser, Cys, Thr, Val, Asn, Leu, Ile, Gln, His) were permitted at position 3 to direct efficient protein N-myristoylation. In this case, the presence of Lys at position 7 was found to affect the amino acid requirement at position 3 and Lys became permitted at this position. When Ser was not located at position 6, only 3 amino acids (Ala, Asn, Gln) were permitted at position 3 to direct efficient protein N-myristoylation. The amino acid requirements found in this study were fully consistent with the N-terminal sequence of 78 N myristoylated proteins in which N-myristoylation was experimentally verified. These observations strongly indicate that the combination of amino acids at position 3, 6 and 7 is a major determinant for protein N-myristoylation. PMID- 14764104 TI - The role and regulation of programmed cell death in plant-pathogen interactions. AB - It is commonly known that animal pathogens often target and suppress programmed cell death (pcd) pathway components to manipulate their hosts. In contrast, plant pathogens often trigger pcd. In cases in which plant pcd accompanies disease resistance, an event called the hypersensitive response, the plant surveillance system has learned to detect pathogen-secreted molecules in order to mount a defence response. In plants without genetic disease resistance, these secreted molecules serve as virulence factors that act through largely unknown mechanisms. Recent studies suggest that plant bacterial pathogens also secrete antiapoptotic proteins to promote their virulence. In contrast, a number of fungal pathogens secrete pcd-promoting molecules that are critical virulence factors. Here, we review recent progress in determining the role and regulation of plant pcd responses that accompany both resistance and susceptible interactions. We also review progress in discerning the mechanisms by which plant pcd occurs during these different interactions. PMID- 14764105 TI - Integration of environmental and host-derived signals with quorum sensing during plant-microbe interactions. AB - Many plant-associated microbes use secreted autoinducer molecules, including N acylhomoserine lactones (AHLs), to regulate diverse behaviours in association with their population density (quorum sensing). Often, these responses are affected by environmental conditions, including the presence of other AHL producing bacterial species. In addition, plant-derived metabolites, including products that arise as a direct result of the bacterial infection, may profoundly influence AHL-regulated behaviours. These plant products can interact directly and indirectly with the quorum-sensing network and can profoundly affect the quorum-sensing behaviour. Local conditions on a microscopic scale may affect signal molecule longevity, stability and accumulation, and this could be used to give information in addition to cell density. Furthermore, in many Gram-negative bacteria, AHL signalling is subservient to an additional two-component signalling system dependent upon homologues of GacS and GacA. The signal(s) to which GacS responds are not known, but recent research suggests that a self-produced ligand may be being detected. This review will focus on two well-studied examples of AHL regulated plant-associated behaviour, Erwinia carotovora and Agrobacterium tumefaciens, to illustrate the complexity of such signalling networks. PMID- 14764106 TI - Systemic cytokine response in murine anthrax. AB - Systemic pro-inflammatory cytokine release has been previously implicated as a major death-causing factor in anthrax, however, direct data have been absent. We determined the levels of IL-1 beta, IL-6 and TNF-alpha in serum of mice challenged with virulent (Ames) or attenuated (Sterne) strains of Bacillus anthracis. More than 10-fold increase in the IL-1beta levels was detected in Ames challenged Balb/c mice, in contrast to more susceptible C57BL/6 mice, which showed no IL-1beta response. Balb/c mice have also responded with higher levels of IL-6. The A/J mice demonstrated IL-1beta and IL-6 systemic response to either Ames or Sterne strain of B. anthracis, whereas no increase in TNF-alpha was detected in any murine strain. We used RT-PCR for gene expression analyses in the liver which often is a major source of cytokines and one of the main targets in infectious diseases. A/J mice challenged with B. anthracis (Sterne) showed increased gene expression for Fas, FasL, Bax, IL-1 beta, TNF-alpha, TGF-beta, MIP 1alpha, KC and RANTES. These data favour the hypothesis that apoptotic cell death during anthrax infection causes chemokine-induced transmigration of inflammatory cells to vitally important organs such as liver. Administration of caspase inhibitors z-VAD-fmk and ac-YVAD-cmk improved survival in Sterne-challenged mice indicating a pathogenic role of apoptosis in anthrax. PMID- 14764107 TI - Characterization of flagellin expression and its role in Listeria monocytogenes infection and immunity. AB - Flagellin is the structural component of flagella produced by many pathogenic bacteria and is a potent proinflammatory molecule that mediates these effects through Toll-like receptor (TLR) 5. In Listeria monocytogenes (LM), flagellin expression is regulated by temperature and has been described as being shut off at 37 degrees C. In this study, we demonstrate that TLR5-mediated cell activation and flagellin expression is maintained at 37 degrees C in some laboratory-adapted strains and in approximately 20% of LM clinical isolates. To determine the role of flagellin in LM infection, a targeted mutation in the structural gene for flagellin (flaA) was generated in a parental LM strain that expressed flagellin under all conditions examined. In vitro studies demonstrated that this deltaflaA mutant was (i). non-motile; (ii). not able to activate TLR5-transfected HeLa cells; and (iii). induced tumour necrosis factor (TNF)-alpha production in approximately 50% fewer CD11b+ cells in splenocytes from normal mice compared with the parental strain. However, there was no significant alteration in virulence of the deltaflaA mutant after either intravenous or oral murine infection. Similarly, there was no difference in the generation of LM-specific CD8 or CD4 T cells after intravenous or oral infection. These data indicate that flagellin is not essential for LM pathogenesis or for the induction of LM specific adaptive immune responses in normal mice. PMID- 14764108 TI - Enterohaemorrhagic and enteropathogenic Escherichia coli use different mechanisms for actin pedestal formation that converge on N-WASP. AB - Enteropathogenic Escherichia coli (EPEC) and enterohaemorrhagic E. coli (EHEC), two closely related diarrhoeagenic pathogens, induce actin rearrangements at the surface of infected host cells resulting in the formation of pseudopod-like structures termed pedestals beneath intimately attached bacteria. We have shown previously that N-WASP, a key integrator of signalling pathways that regulate actin polymerization via the Arp2/3 complex, is essential for pedestal formation induced by EPEC using N-WASP-defective cell lines. Here we show that actin pedestal formation initiated by EHEC also depends on N-WASP. Amino acid residues 226-274 of N-WASP are both necessary and sufficient to target N-WASP to sites of EHEC attachment. The recruitment mechanism thus differs from that used by EPEC, in which amino-terminal sequences of N-WASP mediate recruitment. For EPEC, recruitment of N-WASP downstream of Nck has been postulated to be mediated by WIP. However, we find a direct interaction of N-WASP with WIP to be dispensable for EPEC-induced pedestal formation and present data supporting an F-actin dependent localization of WIP to actin pedestals induced by both EPEC and EHEC. In summary, our data show that EPEC and EHEC use different mechanisms to recruit N-WASP, which is essential for actin pedestal formation induced by both pathogens. PMID- 14764109 TI - Helicobacter pylori gamma-glutamyltranspeptidase upregulates COX-2 and EGF related peptide expression in human gastric cells. AB - Gastric mucosa responds to Helicobacter pylori-induced cell damage by increasing the expression of COX-2 and EGF-related peptides. We sought to investigate the bacterial virulence factor/s and the host cellular pathways involved in the upregulation of COX-2, HB-EGF and amphiregulin in MKN 28 and AGS gastric mucosal cells. H. pylori strain CCUG 17874 was grown in Brucella broth supplemented with 0.2% (2,6-dimethyl)-beta-cyclodextrins. The soluble proteins released in the culture medium by the bacterium were fractionated by exclusion size and anion exchange chromatography. A single peak retaining the ability to upregulate COX-2 and HB-EGF mRNA and protein expression was obtained. SDS-PAGE analysis of the peak showed two peptides with an apparent molecular weight of 38 and 22 kDa, which were identified by automated Edman degradation analysis as the N-terminal and C-terminal peptides of H. pylori gamma-glutamyltranspeptidase respectively. Acivicin, a selective gamma-glutamyltranspeptidase inhibitor, counteracted H. pylori-induced upregulation of COX-2 and EGF-related peptide mRNA expression. An H. pylori isogenic mutant gamma-glutamyltranspeptidase-deficient strain did not exert any effect on COX-2, HB-EGF and amphiregulin mRNA expression. Blockade of phosphatidylinositol-3 kinase and p38 kinase, but not MAP kinase kinase, inhibited H. pylori gamma-glutamyltranspeptidase-induced upregulation of COX-2 and EGF-related peptide mRNA expression. PMID- 14764110 TI - Polysaccharide intercellular adhesin (PIA) protects Staphylococcus epidermidis against major components of the human innate immune system. AB - The skin commensal and opportunistic pathogen Staphylococcus epidermidis is the leading cause of nosocomial and biofilm-associated infections. Little is known about the mechanisms by which S. epidermidis protects itself against the innate human immune system during colonization and infection. We used scanning electron microscopy to demonstrate that the exopolysaccharide intercellular adhesin (PIA) resides in fibrous strands on the bacterial cell surface, and that lack of PIA production results in complete loss of the extracellular matrix material that has been suggested to mediate immune evasion. Phagocytosis and killing by human polymorphonuclear leucocytes was significantly increased in a mutant strain lacking PIA production compared with the wild-type strain. The mutant strain was also significantly more susceptible to killing by major antibacterial peptides of human skin, cationic human beta-defensin 3 and LL-37, and anionic dermcidin. PIA represents the first defined factor of the staphylococcal biofilm matrix that protects against major components of human innate host defence. PMID- 14764111 TI - Production of phthiocerol dimycocerosates protects Mycobacterium tuberculosis from the cidal activity of reactive nitrogen intermediates produced by macrophages and modulates the early immune response to infection. AB - The growth of Mycobacterium tuberculosis mutants unable to synthesize phthiocerol dimycocerosates (DIMs) was recently shown to be impaired in mouse lungs. However, the precise role of these molecules in the course of infection remained to be determined. Here, we provide evidence that the attenuation of a DIM-deficient strain takes place during the acute phase of infection in both lungs and spleen of mice, and that this attenuation results in part from the increased sensitivity of the mutant to the cidal activity of reactive nitrogen intermediates released by activated macrophages. We also show that the DIM-deficient mutant, the growth and survival of which were not impaired within resting macrophages and dendritic cells, induced these cells to secrete more tumour necrosis factor (TNF)-alpha and interleukin (IL)-6 than the wild-type strain. Although purified DIM molecules by themselves had no effect on the activation of macrophages and dendritic cells in vitro, we found that the proper localization of DIMs in the cell envelope of M. tuberculosis is critical to their biological effects. Thus, our findings suggest that DIM production contributes to the initial growth of M. tuberculosis by protecting it from the nitric oxide-dependent killing of macrophages and modulating the early immune response to infection. PMID- 14764112 TI - Interaction of Shiga toxin from Escherichia coli with human intestinal epithelial cell lines and explants: Stx2 induces epithelial damage in organ culture. AB - Shiga toxins (Stx) produced by Escherichia coli are associated with systemic complications such as haemolytic-uraemic syndrome. The mechanism of Stx translocation across the epithelial barrier is unknown as human intestinal epithelium lacks receptor Gb3. In this study, we have examined the interaction of purified Stx1 and 2 with Caco-2 (Gb3+) and T84 (Gb3-) cell lines, and determined the effects of Stx on human intestine using in vitro organ culture (IVOC). Stx exposure caused inhibition of protein synthesis and apoptosis in Caco-2 but not in T84 cells. However, both Stx1 and 2 were transported to the endoplasmic reticulum, and the Stx1 A-subunit was cleaved in a furin-dependent manner in both cell lines. Thus, a Gb3-independent retrograde transport route exists in T84 cells for Stx that does not induce cell damage. IVOC demonstrated increased epithelial cell extrusion in response to exposure to Stx2, but not Stx1, in both small intestine and colon. Pretreatment of Stx2 with Stx2-specific antibody abrogated this effect. Overlaying frozen sections with Stx showed lamina propria, but not epithelial, cell binding that paralleled Gb3 localization, and included endothelium and pericryptal myofibroblasts. This indicates that human intestinal epithelium may evince Stx2-induced damage in the absence of Gb3 receptors, by an as yet unrecognized mechanism. PMID- 14764113 TI - Robotics in urology. PMID- 14764114 TI - The antegrade continence enema procedure. PMID- 14764115 TI - Should urologists do their own diagnostic ultrasonography? PMID- 14764116 TI - Counting the cost of treating benign prostatic hyperplasia. PMID- 14764117 TI - Renal autotransplantation. PMID- 14764118 TI - Intermittent androgen suppression. PMID- 14764119 TI - The development of laser prostatectomy. PMID- 14764120 TI - Managing patients after an ileal orthotopic bladder substitution. PMID- 14764121 TI - The effect of sampling more cores on the predictive accuracy of pathological grade and tumour distribution in the prostate biopsy. AB - OBJECTIVE: To determine if increasing the number of cores at biopsy improves the predictive accuracy of the Gleason score or aids in anticipating the location and volume of prostate tumour. PATIENTS AND METHODS: The charts of 75 consecutive patients who underwent radical retropubic prostatectomy for clinical T1-2 adenocarcinoma of the prostate were reviewed retrospectively; 31 patients had a sextant biopsy (group 1) and 44 had > or = 8 cores taken (group 2). The concordance between biopsy data and final prostatectomy Gleason score, tumour location and volume was determined for each group. RESULTS: There were no differences in mean age, prostate-specific antigen level before biopsy or biopsy Gleason score for the two groups; 58% of group 1 had their final pathological grade changed after prostatectomy, vs 29% of group 2 (P < 0.05). In neither group was there a significant correlation between the percentage of cores positive for tumour and the percentage volume of prostate involved with cancer, or the ability of the biopsy to predict tumour location. CONCLUSION: Taking > or = 8 biopsy cores improved the pathological grading accuracy, which may be valuable in choosing a treatment for the patient with newly diagnosed prostate cancer. PMID- 14764122 TI - A pilot study of use of the cyclooxygenase-2 inhibitor celecoxib in recurrent prostate cancer after definitive radiation therapy or radical prostatectomy. AB - OBJECTIVES: To evaluate the efficacy of the cyclooxygenase (COX)-2 inhibitor celecoxib in prostate-specific antigen (PSA) recurrent prostate cancer after definitive radiation therapy (RT) or radical prostatectomy (RP), as recent evidence showed that COX-2 inhibitors have potent antitumour activity in prostate cancer both in vitro and in vivo but there are no human trials. PATIENTS AND METHODS: Twelve patients who had biochemical relapse after RT or RP were treated with celecoxib 200 mg twice daily. Follow-up PSA levels to assess efficacy were obtained at 3, 6 and 12 months after initiating treatment. Data were evaluated by calculating PSA doubling times and the slope of the curve of logPSA vs time, to assess rate of PSA rise before and after celecoxib treatment for each patient. Serum testosterone levels were also measured. RESULTS: Eight of the 12 patients had significant inhibition of their serum PSA levels after 3 months of treatment; five had a decline in their absolute PSA level and three a stabilization of the level. Of the remaining four patients, three had a marked decrease in their PSA doubling time, with a mean increase (i.e. slowing) of 3.1 times that before treatment. The short-term responses at 3 months also continued at 6 and 12 months. From the slope of log PSA vs time there was a significant flattening of the rate of PSA rise (P = 0.001). There was a significant change of patients with rapid doubling times towards slower doubling times or even stable/declining PSA values after treatment with celecoxib (P = 0.029). There was no significant change in testosterone levels, suggesting an androgen-independent mechanism. CONCLUSIONS: COX-2 inhibitors may have an effect on serum PSA levels in patients with biochemical progression after RT or RP. These results suggest that COX-2 inhibitors may help to delay or prevent disease progression in these patients, and thereby help extend the time until androgen deprivation therapy. Further study with more patients is currently underway to better evaluate the clinical potential of COX-2 inhibitors as an antitumour agents in prostate cancer. PMID- 14764123 TI - A low incidence of positive surgical margins in prostate cancer at high risk of extracapsular extension after a modified anterograde radical prostatectomy. AB - OBJECTIVES: To evaluate the incidence of positive surgical margins (and associated risk factors) in patients with localized prostate cancer at high preoperative risk of extracapsular disease treated using a modified anterograde radical retropubic prostatectomy technique. Positive surgical margins are an important risk factor for disease recurrence after radical prostatectomy, particularly in patients with extracapsular disease. PATIENTS AND METHODS: In total, 84 patients with clinically localized prostate cancer and a preoperative prostate-specific antigen (PSA) level > 10 ng/mL and/or a biopsy Gleason score > or = 7 were evaluated. The surgical technique allows easy, wide resection of the posterolateral prostatic pedicles, and good mobilization and exposure of the apex before the urethra transection. Prostatectomy specimens were examined for extracapsular tumour spread and positive surgical margins. Differences in putative risk factors (Gleason score, preoperative PSA level, prostate weight) between the positive- and negative-margin groups were evaluated using the Mann Whitney test. RESULTS: Overall, 11 of the 84 (13%) patients had positive surgical margins and of these a single site was involved in six. In total, 15 positive margin sites were identified (five apical, four basal, three posterolateral, two anterior and one posterior). All patients with positive margins had histological extracapsular disease. The preoperative PSA level and Gleason score were significantly higher in the positive- than in the negative-margin group (P = 0.025 and 0.035, respectively). CONCLUSIONS: The anterograde radical prostatectomy minimizes the incidence of positive surgical margins in patients at high risk of extracapsular disease. PMID- 14764124 TI - Incidental carcinoma of the prostate: clinicopathological, stereological and immunohistochemical findings studied with logistic regression and self-organizing feature maps. AB - OBJECTIVE: To identify significant predictive factors determining category T1a and T1b in incidental prostatic carcinoma with classical and neural multivariate data analysis methods. MATERIALS AND METHODS: Incidental prostatic carcinomas diagnosed in our department during 1990-99 (66 cases) were re-examined. Besides acquiring routine clinical and pathological data the tumours were assessed by scoring immunohistochemistry for proliferative activity and p53-overexpression. Tumour vascularization (angiogenesis) and epithelial texture variables were investigated by quantitative stereology. The data were evaluated by classical statistical methods (t-test, correlation analysis, logistic regression). Moreover, self-organizing feature maps (SOMs) were applied as an exploratory approach to unsupervised data analysis by artificial neural networks. RESULTS: The proliferative fraction, p53 overexpression of tumour cell nuclei, preoperative prostate-specific antigen value and density of capillary vascularization correlated with the Gleason score in incidental prostatic carcinoma. In a stepwise logistic regression analysis with the tumour categories T1a and T1b as dependent variables, the Gleason score and the volume fraction of epithelial cells were significant independent predictors of the tumour category. The cases could be grouped into clusters of different degrees of malignancy using SOMs. CONCLUSIONS: Texture variables of tumour cells are of central importance for the extent of propagation in the prostate in incidental prostatic adenocarcinomas. Gleason score and quantitative stereological estimates of the volume fraction of tumour cells are significant predictors of T1a and T1b categories of incidental prostatic carcinoma. Unsupervised clustering of T1 prostate carcinoma cases by SOMs correlates well with the dichotomous classification into T1a and T1b according to the UICC. PMID- 14764125 TI - Increased expression of caveolin-1 and microvessel density correlates with metastasis and poor prognosis in clear cell renal cell carcinoma. AB - OBJECTIVE: To investigate the relationship of caveolin-1 expression and microvessel density (MVD), a reflection of angiogenesis, with metastasis and prognosis in patients with clear cell renal cell carcinoma (RCC). PATIENTS AND METHODS: Formalin-fixed, paraffin-embedded tissue sections of clear cell RCC from 67 patients who had undergone radical nephrectomy were stained immunohistochemically with specific antibodies against caveolin-1 and CD34. Caveolin-1 immunostaining was semi-quantitatively estimated based on the proportion (percentage of positive cells) and intensity. MVD was determined with CD34-stained slides. The expression pattern of caveolin-1 and MVD was compared with the clinicopathological variables. RESULTS: Eighteen patients had either synchronous or metachronous metastases and 11 died during the follow-up. Caveolin 1 intensity was significantly correlated with tumour size (P = 0.005), TNM stage (P = 0.028), M stage (P = 0.012), grade (P = 0.015), and metastasis (synchronous or metachronous; P < 0.001). The caveolin-1 proportion (P = 0.037) and MVD (P = 0.011) were significantly correlated with metastasis. MVD was correlated with caveolin-1 intensity (r = 0.385, P = 0.001) and caveolin-1 proportion (r = 0.388, P = 0.001). There was no difference in the expression of caveolin-1 and MVD between primary and metastatic sites. The survival of patients with higher caveolin-1 intensity was significantly worse than that of patients with lower caveolin-1 intensity. Multivariate analyses indicated that only M-stage was an independent prognostic factor for cancer-specific survival and caveolin-1 expression was not an independent factor. CONCLUSIONS: Increased expression of caveolin-1 and MVD is associated with metastasis and a worse prognosis in clear cell RCC. Caveolin-1 expression is correlated with MVD. These results suggest that caveolin-1 may be important in the progression of clear cell RCC and angiogenesis may be affected by caveolin-1 during the progression of RCC. PMID- 14764126 TI - Expression of vascular endothelial growth factor protein in human renal cell carcinoma. AB - OBJECTIVE: To evaluate the effect of vascular endothelial growth factor (VEGF, one of the most important angiogenetic factors) in renal cell carcinoma (RCC) by analysing many RCCs for the expression of immunohistochemical (IHC) VEGF-staining related to clinicopathological findings and survival. PATIENTS AND METHODS: VEGF immunostaining was examined with the tissue microarray (TMA) method on tumour samples from 229 patients and validated in 71 by ordinary tissue sections (TS). IHC VEGF expression was quantified by estimating the volume density and staining intensity on a three-grade scale. RESULTS: In most RCCs there was VEGF staining in the cell cytoplasm and membrane. In cell membranes the VEGF expression declined with storage time. IHC VEGF expression analysed by TMA and TS gave corresponding results. There was no difference in VEGF expression among conventional, papillary and chromophobe RCCs. There were significant correlations between VEGF expression and tumour size and stage. In univariate analysis VEGF expression correlated with survival, especially in conventional RCCs; this prognostic information was lost in multivariate analysis. The VEGF staining intensity correlated only with VEGF expression but not with any clinicopathological factors. CONCLUSIONS: VEGF protein was present in most RCC cells. There was no difference in VEGF expression among the different RCC types. The correlation between VEGF expression and tumour stage and with prognosis indicates the significance of VEGF within tumour growth and progression in RCC. PMID- 14764127 TI - Randomized, double-blind placebo- and tolterodine-controlled trial of the once daily antimuscarinic agent solifenacin in patients with symptomatic overactive bladder. AB - OBJECTIVE: To assess in a phase 3a trial the efficacy of solifenacin succinate, a once-daily oral antimuscarinic agent in development at 5-mg and 10-mg dosage strengths, for the treatment of overactive bladder (OAB) (Yamanouchi Pharmaceutical Co. Ltd, Tokyo, Japan) compared with placebo in patients with symptoms of OAB, i.e. urgency, incontinence, and frequency, with additional objectives being to assess the safety and tolerability of solifenacin and to compare the efficacy and safety of solifenacin with tolterodine 2 mg twice daily. PATIENTS AND METHODS: The study was an international, multicentre, randomized, double-blind, tolterodine- and placebo-controlled trial conducted at 98 centres. Adult patients with symptomatic OAB for > or = 3 months were eligible; after a single-blind 2-week placebo run-in period patients were randomized equally to a 12-week double-blind treatment with either tolterodine 2 mg twice daily, placebo, solifenacin 5 mg or 10 mg once daily. Efficacy variables included change from baseline in the mean number of urgency, incontinence and urge incontinence episodes, and change from baseline in voids/24 h and mean volume voided/void. RESULTS: In all, 1281 patients were enrolled, 1081 randomized and 1077 treated; 1033 were evaluated for efficacy. Compared with placebo, the change from baseline (-1.41, -32.7%) in the mean number of urgency episodes per 24 h was statistically significantly lower with solifenacin 5 mg (-2.85, -51.9%) and 10 mg (-3.07, 54.7%; both P < 0.001), but not with tolterodine (-2.05, -37.9%; P = 0.0511). There was a statistically insignificant decrease in episodes of incontinence with tolterodine (-1.14; P = 0.1122) but a significant decrease in patients treated with solifenacin 5 (-1.42; P = 0.008) and 10 mg (-1.45; P = 0.0038). Compared with placebo (-1.20, -8.1%) the mean number of voids/24 h was significantly lower in patients receiving tolterodine (-1.88, -15%; P = 0.0145), solifenacin 5 ( 2.19, -17%) and 10 mg (-2.61, -20%; both P < 0.001). The mean volume voided/void was also significantly higher with all three active treatments (P < 0.001). Solifenacin was well tolerated; compared with placebo (4.9%), dry mouth (the most common side-effect), mostly mild, was reported in 18.6% of patients receiving tolterodine, 14.0% receiving 5 mg and 21.3% receiving 10 mg solifenacin. CONCLUSION: Solifenacin 5 and 10 mg once daily improved urgency and other symptoms of OAB, and was associated with an acceptable level of anticholinergic side-effects. Solifenacin demonstrated significantly favourable efficacy to side effect ratio in treating symptomatic OAB. PMID- 14764128 TI - Duloxetine vs placebo in the treatment of stress urinary incontinence: a four continent randomized clinical trial. AB - OBJECTIVES: To further assess, in a phase 3 study, treatment with duloxetine for women with stress urinary incontinence (SUI) in other geographical regions, including Argentina, Australia, Brazil, Finland, Poland, South Africa and Spain, as previous trials in North America and Europe provided evidence for the safety and efficacy of duloxetine as a pharmacological treatment for SUI in women. PATIENTS AND METHODS: The study included 458 women aged 27-79 years enrolled in a double-blind, placebo-controlled trial. The patients with predominantly SUI were identified using a validated clinical algorithm. They were randomly assigned to receive placebo (231) or duloxetine 40 mg twice daily (227) for 12 weeks. The primary outcome variables included the incontinence episode frequency (IEF) and the Incontinence Quality of Life (I-QOL) questionnaire. Van Elteren's test was used to analyse the percentage changes in IEF where the stratification variable was weekly baseline IEF (IEF < 14 and > or = 14). Analysis of covariance was used to analyse I-QOL scores. RESULTS: The mean baseline IEF was 18.4/week; 55% of patients had a baseline IEF of > or = 14. There was a significantly greater median decrease in IEF with duloxetine with placebo (54% vs 40%, P = 0.05), with comparable significant improvements in quality of life (I-QOL score increases of 10.3 vs 6.4, P = 0.007). The improvements with duloxetine were associated with significantly greater increases in voiding intervals than with placebo (20.4 vs 8.5 min, P < 0.001). The placebo response was 10.7% and 12.5% higher than those reported in two European and North American phase 3 trials. This may have been related to more patients being naive for incontinence management in the current trial. Discontinuation rates for adverse events were 1.7% for placebo and 17.2% for duloxetine (P < 0.001), with nausea being the most common reason for discontinuation (3.1%); it was the most common adverse event with duloxetine, but was mild or moderate in most (81%), did not worsen in any patient and resolved within 7 days in 60% and within 1 month in 86% of continuing patients; 88% of women who experienced nausea while taking duloxetine completed the trial. CONCLUSIONS: These results show improvements in incontinence and quality of life with duloxetine 40 mg twice daily for 12 weeks that are in keeping with those reported in two other recently completed phase 3 trials in Europe and North America. PMID- 14764129 TI - Transvaginal electrical stimulation in the treatment of urinary incontinence. AB - OBJECTIVE: To determine the effectiveness of transvaginal electrical stimulation (TES) in treating urinary incontinence, and to assess the clinical improvement 6 months after ending the treatment. PATIENTS AND METHODS: In a double-blind randomized controlled clinical trial, 36 women (24 patients and 12 controls) with stress, urge or mixed urinary incontinence were chosen to use TES or placebo (identical equipment but with no electrical current). The patients had their treatment at home twice a day (20-min sessions) for 12 weeks. They completed a voiding diary and had a urodynamic study at the beginning and end of treatment. They were clinically re-evaluated after 6 months. RESULTS: The mean time of use of TES was similar in both groups (approximately 40 h); the treatment group had a significant increase in maximum bladder capacity (P < 0.02), a significant reduction in the total number of voids (over 24 h; P < 0.02), in the number of episodes of voiding urgency (P < 0.001) and, importantly, in the number of episodes of urinary incontinence (P < 0.001). At the first evaluation, after ending the treatment, 88% of the patients had a significant reduction in symptoms or went into remission. At the 6-month re-evaluation, a third of the patients required another therapeutic approach. CONCLUSION: TES is a practical alternative with few side-effects, and is effective for treating the main forms of female urinary incontinence. PMID- 14764130 TI - The prevalence of urinary incontinence in women in four European countries. AB - OBJECTIVE: To determine the prevalence, type and treatment behaviour of women with urinary incontinence in four European countries. SUBJECTS AND METHODS: Data were collected using a postal survey which was sent to 29,500 community-dwelling women aged > or = 18 years in France, Germany, Spain and the UK. Subjects were asked about the type of urinary incontinence they had experienced and their treatment behaviour. RESULTS: Of the women who responded, 35% reported involuntary loss of urine in the preceding 30 days; stress urinary incontinence was the most prevalent type. The lowest prevalence was in Spain (23%), while the prevalence was 44%, 41% and 42% for France, Germany and the UK, respectively. About a quarter of women with urinary incontinence in Spain (24%) and the UK (25%) had consulted a doctor about it; in France (33%) and Germany (40%) the percentages were higher. Overall, <5% of the women had ever undergone surgery for their condition. While pads were used by half of the women, there were some differences among the countries. CONCLUSIONS: Millions of women in Europe have urinary incontinence; the consultation and treatment rates were low in the European countries included in this study. PMID- 14764131 TI - Criterion validity, test-retest reliability and sensitivity to change of the St George Urinary Incontinence Score. AB - OBJECTIVE: To assess the criterion validity, test-retest reliability and the sensitivity to change after treatment of the St George Urinary Incontinence Score (SGUIS). PATIENTS AND METHODS: Women presenting with urinary incontinence completed two SGUIS tests one week apart. A frequency-volume chart (FVC) was completed before the first attendance and the women had a 1-h pad test before treatment. Patients were treated conservatively under the care of a urogynaecologist or nurse continence advisor, or surgically with an open or laparoscopic colposuspension. After treatment, the SGUIS, FVC and 1-h pad test were repeated. RESULTS: The SGUIS correlated moderately well with the number of leaks per week (Spearman's r = 0.610, 95% confidence interval 0.516-0.689, P < 0.001) but less well with the 1-h pad test loss (r = 0.257, 0.124-0.380, P = 0.002). The test-retest reliability was acceptable, as the mean (SD) difference between the first and second SGUIS was 0.337 (2.675), with limits of agreement of -5.012 to -5.686. The change in the SGUIS after treatment correlated well with the improvement in the number of leaks per week (r = 0.742, 0.662-0.805, 156 samples, P < 0.001) but not as well with the change in 1-h pad test loss (r = 0.531, 0.405-0.636, 151, P < 0.001), although the trend was similar. CONCLUSION: The criterion validity of the SGUIS appeared adequate for both the number of leaks per week on the FVC and leakage on the 1-h pad test. The statistical reproducibility of the test was adequate and appeared more responsive to change after treatment than the other measures. The score is suitable for rapid self administration by patients with a range of incontinence types, unlike most other currently available test instruments. PMID- 14764132 TI - Endothelial nitric oxide synthase expression in neurogenic urinary bladders treated with intravesical resiniferatoxin. AB - OBJECTIVE: To investigate endothelial nitric oxide synthase (eNOS) immunoreactivity in bladder biopsies from patients with neurogenic detrusor overactivity (NDO) before and after treatment with intravesical resiniferatoxin, and compare this with control material; the distribution of two other vascular markers, von Willebrand Factor (vWF) and the vascular endothelial growth factor (VEGF), was also studied. PATIENTS AND METHODS: Flexible cystoscopic bladder biopsies from eight controls investigated for asymptomatic microhaematuria and 19 patients with refractory spinal NDO enrolled in a clinical trial of intravesical treatment with escalating doses of resiniferatoxin were immunostained with polyclonal rabbit antibodies for eNOS, vWF and VEGF. Fewer baseline NDO specimens (eight) were available for vWF and VEGF staining. Computerized image analysis was used to quantify immunoreactivity, and the Mann-Whitney test for statistical analysis. RESULTS: eNOS immunoreactivity was found in the suburothelium and less often in the urothelium, with a distribution indicating a location in small blood vessels at the urothelium-suburothelium junction. Immunostaining for vWF showed a similar location. There was a trend to higher eNOS values before treatment in those responding than in those not responding to resiniferatoxin (P = 0.059), and a significant reduction in eNOS immunoreactivity after successful treatment (P = 0.016). VEGF staining was weaker but there was a significant increase in pretreatment biopsies of responders to resiniferatoxin (P = 0.048). Clinical and histopathology features were similar in both groups. CONCLUSIONS: The trend for higher eNOS expression in patients with NDO who responded to resiniferatoxin suggests that increased vasculature or vasodilatation in the suburothelium may be necessary for successful intravesical treatment. Further studies with more patients are required to confirm this relationship and to examine the mechanisms underlying changes in vasculature with levels of bladder overactivity. PMID- 14764133 TI - Characteristics of primary nocturnal enuresis in adults: an epidemiological study. AB - OBJECTIVE: To evaluate the prevalence and characteristics of primary nocturnal enuresis (PNE) in adults in Hong Kong, as there are currently limited epidemiological data in adults. SUBJECTS AND METHODS: After a telephone survey, 8534 respondents (3996 males and 4538 females) aged 16-40 years were selected for the study and stratified in age groups. The questionnaire used comprised two parts; the first started with questions mainly about the general demographic background to decrease the sensitivity of the study and to establish rapport. The second part was conducted through an automated telephone interview service, with the questions being asked by recorded messages and the respondents then keying in their responses with no need to converse with an interviewer. This part included questions about enuretic symptoms and a subjective assessment of social and psychological effects of bedwetting, and measurements of the individual's self esteem (Rosenberg Self-esteem Scale) and depression (The Centre for Epidemiological Studies Depression Scale). RESULTS: Of the 8534 subjects interviewed, 196 had PNE, giving an overall prevalence of 2.3% (2.7% males and 2.0% females); of these 196, 36 (18.4%) also had daytime urinary incontinence. Hence, 1.9% of adults (2.2% males and 1.7% females) had monosymptomatic PNE. Of these, 53% wet >3 nights/week and 26% wet every night. Prevalence rates remained relatively stable among different age groups, with no apparent trend of a reduction with age. Compared with nonenuretic normal controls, significantly fewer enuretics reached tertiary education (33.4% vs 17.8%, P < 0.01). Bedwetters had a significantly higher incidence of depression and lower self-esteem, and a higher incidence of sleep disturbances than the control group. Among bedwetters, 32-40% felt that there was some effect on their choice of job, work performance and social activities, whilst 23% felt the condition affected their family life and in making friends of either sex. However, there was no significant difference in the marital status. Interestingly, only 34.5% of females and half of males used various methods before bedtime to prevent bedwetting. CONCLUSIONS: Overall, 2.3% of Hong Kong adults aged 16-40 years have persistent PNE. Unlike PNE in early childhood the prevalence remained relatively unchanged with age, suggesting that enuretic symptoms persisting into adulthood are probably less likely to resolve with time. Also, significantly more patients had more severe enuretic symptoms. These findings therefore highlight the possibility that PNE in adults may represent a more pronounced form of the condition, and with a more serious social and psychological effect on affected individuals. Further work is needed to evaluate the pathogenesis and management strategy. PMID- 14764134 TI - A novel temporary prostatic stent for the relief of prostatic urethral obstruction. AB - OBJECTIVES: To determine the effect of a novel temporary prostatic stent (The Spanner, AbbeyMoor Medical, Inc., Minnesota, USA) on variables of voiding function and quality of life among patients with prostatic urethral obstruction. PATIENTS AND METHODS: The stent design is very similar to the proximal 4-6 cm of a Foley catheter; this includes a proximal balloon to prevent distal displacement, a urine port situated cephalad to the balloon, and a reinforced stent of various lengths to span most of the prostatic urethra. There is also a distal anchor mechanism attached by sutures, and a retrieval suture which extends to the meatus and deflates the proximal balloon when pulled. The stent was inserted under topical anaesthesia in 30 patients. The maximum flow rate (Qmax), voided volume (W), postvoid residual (PVR), the International Prostate Symptom Score (IPSS) and stent position were assessed. RESULTS: Stents remained in situ for a mean (range) of 57 (1-98) days. The mean overall Qmax at baseline and after insertion were 8.2 and 11.6 mL/s, representing a 42% improvement (P < 0.001); the respective mean overall Ws were similar, at 219.7 and 221.6 mL (0.9% increase, not significant) and the PVRs were 312.1 and 112.3 mL, representing a 64% decrease (P = 0.004). The overall mean IPSS declined from 22.3 before to 7.1 after insertion, representing a 68% decrease (P < 0.001). There were only minor adverse events. The stability, patency and lack of migration of the device were confirmed radiographically up to 12 weeks of use. CONCLUSIONS: This early study shows that this temporary prostatic stent is easily inserted and removed, remains anchored in position, and significantly improves the Qmax, PVR and IPSS while preserving volitional voiding and continence. PMID- 14764135 TI - Feasibility of open simple prostatectomy with early vascular control. AB - OBJECTIVE: To determine the feasibility of open simple prostatectomy with early vascular control in the treatment of benign prostate hyperplasia (BPH), and thus reduce blood loss. PATIENTS AND METHODS: Thirty-seven patients were reviewed from 1991 to 2002; 34 had a Millin's and three a Freyer's prostatectomy performed by one surgeon, with early vascular control. RESULTS: The mean operative duration was 1.3 h and the mean blood loss 841 mL, with a mean decrease in haemoglobin level of 22 g/L. Six (16%) of the patients received a blood transfusion. The mean weight of the prostate removed was 97.8 g; the duration of catheterization and the hospital stay after surgery were 6.21 and 11.7 days, respectively. There was one (3%) death and one case of pulmonary embolism. Three patients (8%) developed stress incontinence. Two failed to void after surgery and one developed acute retention 3 years later. Five patients developed recurrent obstructive symptoms. CONCLUSION: Open simple prostatectomy with early vascular control reduces the amount of blood loss, rendering it a safe option for treating BPH. PMID- 14764136 TI - Female paraurethral cysts: experience of 25 cases. AB - OBJECTIVE: To report and describe the diagnosis and treatment of female paraurethral cysts. PATIENTS AND METHODS: Twenty-five cases of cysts of the external female genitalia were diagnosed over a 7-year period, using a physical examination, routine blood tests, urine analysis, abdominal ultrasonography and cysto-urethroscopy in all. The cysts were incised, drained and marsupialized; no attempt was made to excise or remove the internal wall of the cyst and it remained in situ. RESULTS: All patients responded to simple marsupialization, with no recurrence of the cyst. CONCLUSION: Cysto-urethroscopy and a limited laboratory and imaging evaluation were enough for the diagnosis. PMID- 14764137 TI - Robot-assisted laparoscopic adrenalectomy: preliminary UK results. AB - OBJECTIVE: To describe the results of our first two cases of laparoscopic adrenalectomy using the da Vinci surgical system (Intuitive Surgical, Inc., Mountain View, CA, USA). PATIENTS AND METHODS: Amongst 75 robot-assisted procedures performed at our institution, two patients underwent robot-assisted laparoscopic adrenalectomy. The set-up time, procedure time, hospital stay, complications and outcomes were recorded. RESULTS: Both operations were completed successfully using the robot; the mean (range) set-up time was 31 (25-37) min and mean procedure time 118.5 (107-130) min. One patient had a postoperative pulmonary embolus and was discharged 5 days after surgery; the second patient was discharged after 3 days. There were no intraoperative complications; both patients were well at the 1-year follow-up. CONCLUSIONS: Robot-assisted laparoscopic adrenalectomy is technically feasible and can be conducted efficiently and safely with the da Vinci surgical system. PMID- 14764138 TI - Ten years' experience of retrograde balloon dilatation of pelvi-ureteric junction obstruction. AB - OBJECTIVE: To analyse the long-term success of retrograde balloon dilatation (RBD) of pelvi-ureteric junction obstruction (PUJO). PATIENTS AND METHODS: The records of 58 adult patients (mean age 48 years, range 18-94) who had RBD between 1990 and 2001 were reviewed retrospectively. All symptomatic patients were included and assessed by dynamic renography. Success was judged as a symptomatic and/or functional improvement. Recurrence of symptoms was recorded and any evidence of deterioration of function despite improved symptoms. Complications were also recorded. RESULTS: In all, 44 (76%) patients were asymptomatic after RBD. There was no relationship between symptomatic change and renographic improvement; some patients who were rendered pain-free had either no improvement or even deterioration in their split renal function. Conversely, some patients who had a demonstrable improvement in their renal function continued to be symptomatic. However, a long-term follow-up (mean 6.1 years) of nine patients who were symptomatically improved showed good maintenance of split renal function. CONCLUSION: RBD is an effective treatment for PUJO and its success is maintained in the long term. PMID- 14764139 TI - Efficacy of extracorporeal shock wave lithotripsy for solitary lower calyceal stone: a statistical model. AB - OBJECTIVE: To evaluate the effect of inferior calyceal radiographic anatomy, number of extracorporeal shock wave lithotripsy (ESWL) sessions and stone size on the successful clearance of solitary inferior calyceal calculi after ESWL. PATIENTS AND METHODS: In a prospective study between January 2001 and November 2002, 66 renal units with a solitary inferior calyceal calculus of < or = 2 cm were treated with electrohydraulic ESWL. The infundibulopelvic angle (two definitions), infundibulovertebral angle, inferior calyceal infundibular diameter, infundibular length, cortical thickness over the lower pole, number of minor calyces and stone size were determined from intravenous urography before treatment. The number of ESWL sessions was also included in the analysis. Treatments which produced residual fragments not clearing within 3 months of satisfactory fragmentation were considered as failures. All patients in whom the treatment failed were treated successfully by percutaneous nephrolithotomy. The data were then analysed using two different statistical methods; first by intravariable differences using the test of proportion (Fisher's test) and then all the variables together using logistic regression. RESULTS: At 3 months 78.8% of the renal units were clear of stone. All intravariable differences were statistically significant except stone size (<1 cm, 1-2 cm). In a multivariate analysis of all variables, only stone size was the most important predictor for successful stone clearance (P = 0.03). CONCLUSIONS: ESWL is the initial treatment of choice in selected patients with inferior calyceal stones. The stone size appears to be the most important predictor for stone clearance. PMID- 14764140 TI - The 'modified prone position': a new approach for treating pre-vesical stones with extracorporeal shock wave lithotripsy. AB - OBJECTIVE: To investigate the utility of a new 'modified-prone' position for treating pre-vesical stones with extracorporeal shock wave lithotripsy (ESWL), usually considered an acceptable and effective treatment for such stones, but for which many different body positions have been used in an attempt to increase its efficacy. PATIENTS AND METHODS: The study included 268 consecutive patients with a solitary pre-vesical stone who underwent ESWL either prone (69) or in the modified-prone position (199) between May 1999 and August 2001. Only those with one stone between the ureteric orifice and 1 cm proximal to the vesico-ureteric junction were included. In each case the stone diameter, days to stone clearance, number of shock waves applied per treatment, and number of sessions required to become stone-free were recorded. If the treatment failed this was also noted. Success rates in the prone and modified-prone groups were compared and analysed to assess which of the variables influenced success with ESWL. RESULTS: After ESWL, 95.5% of the 268 patients were stone-free; the rates in the prone and modified-prone groups were 89.9% and 97.5%, respectively (P = 0.015). The probability of success with ESWL therapy for pre-vesical calculi in modified prone position was about five times (odds ratio 4.56, 95% confidence interval 1.2 17.7) greater than that expected with when prone. The modified-prone position was an independent factor most significantly influencing success with ESWL in these patients. CONCLUSION: The modified-prone position for ESWL is a new and very effective way to treat patients with pre-vesical stones. PMID- 14764141 TI - Orthotopic ileal neobladder: the influence of reservoir volume and configuration on urinary continence and emptying properties. AB - OBJECTIVE: To evaluate the influence of the volume and configuration of the neobladder on urinary continence and reservoir emptying in orthotopic urinary reservoirs using intestinal segments for bladder replacement after radical cystectomy. PATIENTS AND METHODS: Fifty-nine patients who had had a radical cystectomy and urinary reconstruction with an orthotopic ileal neobladder were followed for > or = 1 year; 27 (group 1) had the ileal neobladder created with a shorter intestinal segment (40 cm) in an elongated shape ('J'), and 32 (group 2) had their reservoir made more spherical with a longer ileal loop (60-65 cm). The rates of urinary continence, enuresis, neobladder capacity and postvoid residual urine were evaluated first at 3-6 months and again 1 year after surgery in both groups. RESULTS: At 3-6 months after surgery urinary incontinence and enuresis were more common in group 1, but at 1 year had the same frequency in both groups, at respectively 11% and 44% in group 1, and 13% and 47% in group 2 (P > 0.05). The neobladder capacity and postvoid residual urine were significantly higher in group 2, at > 600 mL and > 100 mL, respectively, in 14% and 14% of the patients in group 1 and 57% and 52% of those in group 2 (P < 0.05). Urinary retention requiring intermittent catheterization did not occur in group 1 but did in 19% of group 2. CONCLUSION: The orthotopic spherical ileal neobladder with a large initial volume is apparently not associated with better continence rates and is prone to developing progressive enlargement, which can lead to neobladder atony and progressive emptying failure, increasing the chance of complete urinary retention. PMID- 14764142 TI - Dorsal tunica albuginea plication to correct congenital and acquired penile curvature: a long-term follow-up. AB - OBJECTIVE: To evaluate the long-term efficacy of a tunica albuginea dorsal plication technique for treating congenital and acquired penile curvature. PATIENTS AND METHODS: We retrospectively evaluated 83 patients (median age 1.8 years) who had their penile curvature corrected surgically using dorsal tunica albuginea plication between 1992 and 2002. The results were evaluated objectively using a pharmacological erection test or subsequently based either on the parents' reports or patients' self-assessment. The median (range) follow-up was 6 (0.7-10) years. RESULTS: Seventy (84%) patients had penile plication as an integral part of hypospadias repair, while the remaining 13 (16%) with a normal urethra had dorsal plication only. Twenty-eight (34%) of the 83 patients had an erection test during a repeat hypospadias repair or closure of a urethrocutaneous fistula; 22 of these had a straight penis, while the remaining six required additional plication for a satisfactory cosmetic outcome. Parents of 45 (54%) children reported that their child had a normal erection with no chordee during the follow-up. Ten (12%) adult patients reported straight erections enabling satisfactory penetration and sensation during sexual intercourse. None of the patients reported penile shortening or erectile dysfunction after surgery, and none had recurrent curvature during the follow-up. There was no difference in the results between patients with congenital or acquired penile curvature. CONCLUSIONS: Dorsal plication of the tunica albuginea is a simple and effective method in the long term for correcting congenital and acquired penile curvature. PMID- 14764143 TI - A preliminary study of natural-fill radiotelemetered ovine fetal cystometry. AB - OBJECTIVE: To determine whether fetal cystometric studies by radiotelemetry are feasible in the fetal lamb, and potentially suitable for chronically monitoring fetal bladder pressures in an experimental fetal model of bladder outlet obstruction (BOO), as in utero BOO (e.g. caused by posterior urethral valves) results in significant postnatal bladder dysfunction but the pathophysiological progression of fetal bladder maldevelopment remains poorly understood. MATERIALS AND METHODS: The procedure required fetal sheep surgery and anaesthesia. Radiotelemetry implants comprised catheters that transmitted pressure fluctuations to an implant body; data were then transmitted using radio frequency to a receiver that passed this information to a computer. Four fetuses were used with different methods of catheter placement to optimize the technique. RESULTS: Recordings were possible in three of the four sheep; during observation there were: (i) quiet periods with no abdominal or bladder pressure rises; (ii) synchronous activity in the bladder and abdomen; and (iii) discriminate activity, associated with intravesical activity only. Four patterns of discriminate bladder activity were observed, defined as void, immature void, staccato activity and 'unstable' type activity. CONCLUSIONS: Radiotelemetry cystometry for long-term monitoring is feasible in the experimental fetus without causing death or morbidity, or inhibiting growth. The method can discriminate reproducible patterns of detrusor activity. Recorded 'voiding' types were consistent between experiments and as reported in other fetal animal studies. PMID- 14764144 TI - Unilateral multicystic dysplastic kidney: experience in children. AB - OBJECTIVES: To report a retrospective study of unilateral multicystic dysplastic kidneys (MCDK) in children, assessing the contralateral kidneys and urinary tract, the functional consequences, and the urological and nephrological management and outcome, as unilateral MCDK is the most common cause of renal cystic disease in children, and malformations of the contralateral urinary tract and kidney (pelvi-ureteric obstruction, megaureter, reflux, renal dysplasia) have been reported. PATIENTS AND METHODS: The study included 97 patients (60 boys, 37 girls) with MCDK seen between 1985 and 1998; 82 were diagnosed in utero by ultrasonography (US). After birth, the diagnosis was verified by US, renal scanning (in 93) or intravenous urography (in four), and 89 (92%) had voiding cysto-urethrography (VCUG). Of the 97 children, 87 (90% had a mean (range) follow up of 44.3 (15-115) months. RESULTS: The MCDK was removed in 17 children; the follow-up of 75 children (five lost to follow-up) showed total involution of the MCDK in 25%, shrinkage in 60% and a stable size in 15%. None had any sign of malignancy. The contralateral kidney showed anomalies in 19 of 97 children (20%); 12 had a dilated renal pelvis (two with megaureter), six had a high echogenicity of the contralateral kidney (one had reflux, and two also pelvic dilatation). In only four of the 89 children was reflux found by VCUG; 16 of the 19 anomalies were detected by US. Five children needed surgery on the contralateral urinary tract (three a pyeloplasty, and one each a pyeloplasty plus ureteroneocystostomy, and an antireflux procedure). Of the contralateral kidneys 43% showed compensatory hypertrophy. There was mild renal insufficiency in three children; renal function seemed to be slightly impaired in many. Five infants had hypertension (four with spontaneous resolution) caused by renal scarring after pyelonephritis or inborn dysplasia of the contralateral kidney. There were symptomatic urinary tract infections in seven children. CONCLUSION: US can be used safely to diagnose unilateral MCDKs and malformations of the contralateral urinary tract and kidney. In cases where US of the dysplastic kidney remains uncertain renal scintigraphy is necessary to detect the lack of renal function. The low rate of reflux makes routine VCUG unnecessary if the contralateral upper urinary tract and kidney appear to be normal on US. Nephrectomy of the dysplastic kidney in typical cases is also unnecessary. A long-term nephro-urological follow up of children with MCDK is recommended. PMID- 14764145 TI - Modulation of autonomous contractile activity in the isolated whole bladder of the guinea pig. AB - OBJECTIVE: To investigate the actions of the nonhydrolysable analogue of ATP, alpha,beta-methylene ATP (alpha,beta-MATP) and the sensory peptide, substance P, on the phasic activity generated by muscarinic stimulation in the isolated whole bladder. Isolated bladder can generate complex contractions resulting in phasic rises in intravesical pressure (the autonomous bladder): activity thought to underlie nonmicturition activity in vivo and which may be important in generating bladder sensations. MATERIALS AND METHODS: Experiments were conducted on whole isolated bladders from female guinea pigs (270-300 g). Bladders were cannulated via the urethra, suspended in a heated chamber containing oxygenated solution at 33-36 degrees C and intravesical pressure recorded. All drugs were added to the solution bathing the abluminal surface of the bladder. RESULTS: When alpha,beta MATP (30-3000 nmol/L) or substance P (30-300 nmol/L) was added to resting bladders there were small rises in intravesical pressure (<2 cmH2O). However, in the presence of phasic activity generated by exposing the bladder to the muscarinic agonist arecaidine (100-300 nmol/L) or the nicotinic ligand lobeline (10-30 micromol/L) similar or lower concentrations of alpha,beta-MATP or substance P produced more dramatic effects: alpha,beta-MATP and substance P (both at 100 nmol/L) activated a rise in basal pressure of > 15 cmH2O and increased the frequency of the phasic activity. On removing alpha,beta-MATP or substance P, there was a slowing of phasic activity indicative of an inhibitory mechanism. CONCLUSION: In addition to direct effects on smooth muscle the agonists alpha,beta-MATP and substance P appear to be potent regulators of the mechanisms generating phasic activity. A developing concept is that the mechanisms responsible for generating phasic activity underlie nonmicturition activity are the target for excitatory and inhibitory inputs. Regulating such activity may be a factor in generating or modifying bladder sensation. Inappropriate or exaggerated phasic activity could underpin the pathological changes which cause the overactive bladder, thus adding another hypothesis to the neurogenic and myogenic hypotheses of bladder overactivity, i.e. that of the autonomous bladder. PMID- 14764146 TI - Noradrenaline inhibits autonomous activity in the isolated guinea pig bladder. AB - OBJECTIVE: To investigate the actions of noradrenaline and the specific alpha adrenergic agonists cirazoline (alpha1) and UK14304 (alpha2), and beta-receptor agonists formoterol (beta2) and BRL37344 (beta3) on the phasic activity induced by muscarinic stimulation on the isolated guinea pig bladder, as the physiological significance of this activity is unknown but it may underlie non micturition contractions (NMCs, which can be inhibited by sympathetic nerve stimulation) and the generation of bladder sensations. MATERIALS AND METHODS: All experiments were conducted using whole isolated bladders from female guinea pigs (270-300 g). Bladders were cannulated via the urethra and suspended in a heated chamber containing oxygenated Tyrode's solution at 33-35 degrees C and the intravesical pressure recorded. All drugs were added to the solution bathing the abluminal surface. RESULTS: Exposure to noradrenaline reduced the amplitude and frequency of the phasic activity. When noradrenaline was washed off there was a transient increase in frequency. There was marked desensitization with repeated applications of noradrenaline. Applying the specific beta3-agonist BRL37344 reduced the amplitude of the phasic activity while formoterol, a specific beta2 agonist, had no effect. Cirazoline, a specific alpha1-agonist, reduced the amplitude of the responses and significantly reduced the frequency of the phasic activity. UK14304, a specific alpha2-agonist, had no effect. Stimulation of the hypogastric nerve to the guinea pig bladder generates contractions. Prolonged nerve stimulation at low frequency (6.5 Hz) generated phasic rises in intravesical pressure which were inhibited by noradrenaline. Using short (5 s) periods of stimulation noradrenaline inhibited nerve-mediated contractions at all frequencies but was more effective at <10 Hz. CONCLUSION: These experiments show that sympathomimetic stimulation in the isolated whole bladder results primarily in an inhibition of phasic activity, but also a stimulation. Two receptor subtypes appear to be involved in the inhibition, alpha1 and beta3, suggesting that there may be many sites of action. These results are discussed in terms of the possible physiological significance of phasic activity and the potential importance of its inhibition, in the context of the causes of pathological changes in the bladder, particularly those associated with bladder overactivity, and the pharmacological approach to the alleviation of clinical symptoms. PMID- 14764147 TI - Prostate apoptosis after doxazosin treatment in the spontaneous hypertensive rat model. AB - OBJECTIVE: To validate the spontaneous hypertensive rat (SHR) model for basic research into benign prostate hyperplasia (BPH), and to assess doxazosin-induced changes in prostatic structure and apoptotic status. MATERIALS AND METHODS: Four groups of rats were assessed: group 1, 15 SHRs treated with doxazosin; group 2, 14 SHRs with unilateral excision of the major pelvic ganglion; group 3, 14 untreated SHRs; and group 4, 16 intact Wistar-Kyoto (WKY) rats. The doxazosin mesylate (0.03 mg daily) was given compacted in rat food for 3 months. The prostatic ventral lobe (VL) was excised and weighed. Stereological light microscopy, multiplex reverse transcription-polymerase chain reaction of prostate caspases, and caspase-3 activity (cellular enzymatic assay) were assessed. RESULTS: There was more development of the glandular epithelium (P < 0.001) in SHR rats than in controls, which was even greater after doxazosin exposure (P = 0.027). SHR animals had higher caspase expression (P < 0.05) and activity (P = 0.008) than WKY rats, but both were reduced after doxazosin therapy (P < 0.01 and 0.028, respectively). CONCLUSIONS: This study confirmed prostate hyperplasia in the SHR model. Doxazosin exposure did not reduce the volume of glandular epithelium and contributed to protecting against caspase-induced apoptosis. The SHR model may be not a valid option to study doxazosin-induced apoptosis in BPH. PMID- 14764148 TI - Hormone-based approaches: altering the balance. PMID- 14764149 TI - Laparoscopic adrenalectomy with simultaneous management of renal pathology. PMID- 14764150 TI - Safe control of the renal vein during laparoscopic nephrectomy using the 'loop around the vein' technique. PMID- 14764151 TI - Sexual factors and prostate cancer. PMID- 14764152 TI - The future in the past, or the management of invasive prostatic cancer in the Third World. PMID- 14764153 TI - A study to detect Helicobacter pylori in fresh and archival specimens from patients with interstitial cystitis, using amplification methods. PMID- 14764154 TI - Screening for prostate cancer: have you had your cholesterol measured? PMID- 14764155 TI - Identifying under-performing surgeons. PMID- 14764156 TI - Bilateral testicular cancer: a preventable problem? Experience from a large cancer centre. PMID- 14764157 TI - No-scalpel vasectomy: a cautionary tale of failure. PMID- 14764158 TI - Partner's influence on patient preference for treatment in early prostate cancer. PMID- 14764159 TI - Surgical atlas: Radical perineal prostatectomy. PMID- 14764160 TI - Diary. PMID- 14764162 TI - I hope to make the Journal even more successful during 2004. PMID- 14764161 TI - Abbreviations. PMID- 14764163 TI - When u say "U Waves," what do u mean? PMID- 14764164 TI - Dynamics of T-U wave in patients with idiopathic ventricular tachycardia originating from the right ventricular outflow tract. AB - Postextrasystolic U wave augmentation is observed in patients with long QT syndrome and those with organic heart disease. This phenomenon is considered a marker of increased risk of arrhythmia. However, the characteristics of the U wave have not been evaluated in patients with idiopathic VT originating from the right ventricular outflow tract (RVOT-VT). The present study evaluated the dynamic change in the T-U wave in patients with RVOT-VT. Holter ECGs obtained from 14 patients with RVOT-VT and 11 healthy control subjects were analyzed. The amplitude of T and U waves (Tamp and Uamp) and preceding RR intervals were measured during stable sinus rhythm (rate dependent change) and in the postextrasystolic sinus complex (pause dependent change). Uamp correlated negatively and significantly with the preceding RR interval in 13 (93%) RVOT-VT patients but in only 2 (18%) control subjects. The average value of the slope of the Uamp/RR relationship was negative (-0.22 +/- 0.10 mV/s) in the RVOT-VT group, but was positive (0.04 +/- 0.07 mV/s, P < 0.001) in the control group. Pause dependent U wave augmentation was observed in 12 (86%) of 14 patients. Increased frequency of consecutive preceding premature ventricular contractions (PVCs) was associated with a larger postextrasystolic Uamp. PVC or the first ventricular beat of VT arose from near the peak of augmented U waves. The dynamic changes in the T-U wave were observed in patients with RVOT-VT. Further investigations are required to elucidate the precise role of the U wave in arrhythmogenesis in those patients. PMID- 14764166 TI - Ventricular evoked response as clinical marker for hemodynamic changes in dilative cardiomyopathy. AB - The potential value of ventricular evoked response (VER) evaluation by implantable pacemakers as clinical marker for disease induced hemodynamic changes in the heart, has so far not been explicitly evaluated. We conducted a study to evaluate the reproducibility of the R spike and T wave measurements (R(VER) and T(VER)) under controlled clinical conditions and examine the correlation between VER parameters and standard echocardiographic measurements in the left ventricle. Additionally, the utility of the VER as a marker for NYHA classification and the presence of cardiomyopathy was investigated. The Physios CTM 01 pacemaker capable of recording authentic VER signal morphology, was used with low polarization fractal coated pacing leads to obtain high-fidelity VER recordings in 26 patients with conventional pacing indications (mean age: 69.1 +/- 11.8 years; 20 men). Three patients suffered from dilative cardiomyopathy (DCM), 14 from hyperthropic nonobstructive cardiomyopathy (HNCM), and nine had no myopathy but suffered from coronary artery disease (CAD). Five patients were in NYHA Class IV, 19 in Class III, and two in Class II. Mean R(VER) and T(VER) amplitudes were calculated from one-minute VER recordings. Standard echocardiography parameters were determined during this recording. Two follow-ups at a mean distance of 11.3 +/- 5.7 month were performed. The reproducibility of R(VER) or T(VER) (correlation factors: 0.992 and 0.981, respectively) was superior to the reproducibility of any echocardiographic parameter (correlation factors 0.404-0.943). There was no strong correlation between VER and any echo parameter. Both R(VER) and T(VER) were significantly reduced in NYHA Class IV patients (P < 0.05), and nearly significantly reduced in DCM versus other patients (P = 0.05-0.09). HNCM made no difference to CAD. The investigation shows that analysis of VER parameters bears a promising potential for dynamic monitoring of diseases affecting the hemodynamics, and of therapeutic effects, by means of regular, nonburdening pacemaker follow-up examinations. PMID- 14764165 TI - T wave spectral variance for noninvasive identification of patients with idiopathic dilated cardiomyopathy prone to ventricular fibrillation even in the presence of bundle branch block or atrial fibrillation. AB - Conventional methods using Holter ECG recordings for noninvasive risk stratification are limited in patients with idiopathic dilated cardiomyopathy (IDC) prone to ventricular fibrillation (VF) having atrial fibrillation (AF) or bundle branch block (BBB). We therefore investigated, whether spectral assessment of beat-to-beat alternations of repolarization is associated with VF in these patients. Twenty-four-hour Holter ECG recordings in 462 patients with IDC were used. The VF group comprised of 64 consecutive patients who survived cardiac arrest, the no VF group consisted of 398 consecutive patients without a history of malignant ventricular arrhythmia. One hundred patients with ischemic cardiomyopathy (ICM) served as a control group. In each patient, 1,024 consecutive T waves were aligned using cross correlation methods. Two-dimensional Fourier transform (2D FFT) used the data matrix of 1,024 consecutive 200-ms segments centered to the T wave peak. Power spectra of the 2D FFT revealed the frequency content of the T wave in the first dimension and the periodicity of this frequency content in the second dimension. The ratio between periodic frequency contents and the sum of nonperiodic and periodic frequency contents between 0.5 and 50 Hz is equal to the T wave spectral variance (TWSV) index. Thus, TWSV index = 0 would mean that all 1,024 T waves are identical and TWSV index = 1 would mean that the 1,024 T waves are totally variable. The TWSV index was significantly higher in the VF group (0.93 +/- 0.14) than in the no VF group (0.53 +/- 0.13, P < 0.01). The best cutoff between the VF and the no VF group was achieved by using a TWSV index of 0.75 (sensitivity = 89%, specificity = 78%). No significant differences were observed between patients with and without AF or with and without BBB, and between patients with IDC and ICM. Even in the presence of BBB or AF spectral assessment of T wave alternations by TWSV index using 2D FFT in Holter ECG recordings, allows the identification of patients with IDC at risk for VF. PMID- 14764167 TI - Does sinus rhythm beget sinus rhythm? Long-term follow-up of the patient activated atrial defibrillator. AB - The aim of this study was to determine the effect of early patient activated cardioversion of atrial fibrillation (AF) using the atrial defibrillator on recurrence of AF. Fifteen patients, mean age 63 +/- 14 years, 80% men, with drug resistant persistent AF were implanted with the Jewel AF atrial defibrillator. All patients performed self-administered cardioversion for AF recurrences. Over a 2 year follow-up, 238 patient-activated cardioversions were performed in 14 patients. Sinus rhythm was restored on every occasion with 96% of episodes terminating with a single shock. The median time from AF onset to patient awareness of symptoms was 2.5 hours. The median time from onset of symptoms to cardioversion was 3.5 hours. Comparison of the first and second six month period following implant showed a nonsignificant increase in mean total AF duration (75.7 +/- 107.8 hours vs 146.6 +/- 194.1 hours, P = 0.28). Two patients (13%) had a decreasing frequency of AF recurrences. The majority continued to have regular recurrences of AF. The atrial defibrillator is an extremely effective method of restoring sinus rhythm in patients with persistent AF. Regular early use of the atrial defibrillator, increased the duration of sinus rhythm in a minority of patients during long-term follow-up. Most patients had regular recurrences of AF requiring patient-activated cardioversion. PMID- 14764168 TI - ECG criteria for localizing the pulmonary vein origin of spontaneous atrial premature complexes: validation using intracardiac recordings. AB - We have shown that pacemapping from each of the pulmonary veins reveals unique surface ECG characteristics. However, application of these criteria to spontaneous atrial premature complexes is often difficult because of obscuration by the prior T wave. We hypothesized that the pulmonary vein of origin of spontaneous atrial premature complexes can be determined by measuring characteristics of the P wave whether or not the P wave was superimposed on the prior T wave. We analyzed 58 spontaneous atrial premature complexes of known pulmonary vein origin in 30 patients referred for atrial fibrillation ablation. The origin of all the atrial premature complexes was documented by detailed, intracardiac multipolar catheter mapping. Based on previous work, the criteria for distinguishing right-sided from left-sided pulmonary vein origin of atrial premature complex includes: (1) P wave duration < 120 ms; (2) P wave amplitude in lead I > 0.05 mV; and (3) P wave amplitude in leads II/III > 1.25. The criteria to separate superior from inferior pulmonary veins included the sum of the P wave amplitude in all the inferior leads greater than 0.3 mV. The combination of the P wave duration < 120 ms and the ratio of the P wave amplitude in leads II/III > 1.25, distinguished right-sided from left-sided pulmonary vein origin of spontaneous atrial premature complexes with a sensitivity of 82% and specificity of 100%. The sum of the P wave amplitude in leads II, III, and aVF > 0.3 mV distinguished superior from inferior pulmonary vein of origin with a sensitivity of 39% and specificity of 73%. The pulmonary vein origin of spontaneous atrial premature complexes can often be localized using careful quantitative analysis of the surface ECG despite superimposition of the P wave upon the T wave. Separation of right-sided from left-sided pulmonary vein origin of spontaneous atrial premature complexes can be determined with good specificity and sensitivity, while the ability to distinguish inferior from superior pulmonary vein origin is limited. PMID- 14764169 TI - Localization of accessory pathways by the electrocardiogram: which is the degree of accordance of three algorithms in use? AB - We evaluated the extent of agreement among three algorithms used for the localization of accessory pathways in patients with overt preexcitation. By the use of one algorithm, three independent couples of observers localized the accessory pathway in 95 consecutive patients showing overt preexcitation in the 12-lead surface electrocardiogram. We defined the following regions: Left atrioventricular ring (LAVR), Right atrioventricular ring (RAVR), Left lateral/left anterolateral (LL/LAL), Left posterior/left posterolateral (LP/LPL), Left posteroseptal (LPS), Right midseptal (RMS), Right posteroseptal (RPS), Right posterior/right posterolateral (RP/RPL), Right lateral/right anterolateral (RL/RAL), and Right anterior/right anteroseptal (RA/RAS). The extent of agreement in each region was evaluated and compared with the expected one, as calculated from the reported. The extent of agreement was as expected: (1) high in the regions LAVR, RAVR, LL/LPS and (2) limited in the regions LPS, RPS, and (3) clearly lower than expected in the regions LP/LPL, RA/RAS, RMS, RL/RAL. In cases with total or partial disagreement, the number of electrocardiograms with duration of QRS complex smaller than 120 ms was greater than in cases with total agreement (30/46 vs 22/50, P < 0.05). The observed agreement among algorithms is clearly lower than the expected one. Minimal preexcitation, limited number of patients, and arbitrarily defined regions were possibly the reasons for some unexpected results. PMID- 14764170 TI - Electrophysiological evaluation of the sinus node and the cardiac conduction system following the maze procedure for atrial fibrillation. AB - Transient sinus node dysfunction has been demonstrated by noninvasive methods following the maze procedure for atrial fibrillation (AF). However, extensive data from invasive electrophysiological studies have not been previously reported. Thirty-seven patients, mean age 54 +/- 10 years, underwent the maze (III) procedure. Electrophysiological studies with recordings of SNRT, CSNRT, AVN ERP, point of Wenckebach block, AH, PA, and HV interval, were performed preoperatively and 6 and 15 months postoperatively. Induction of atrial flutter/AF was attempted postoperatively. Based on electrophysiological study evaluation, the maze (III) procedure did not cause permanent damage to the sinus node in any patient with a documented normal sinus node function preoperatively (CSNRT max 541 +/- 210 vs 587 +/- 437 ms, P = 0.26). Postoperative AV node function was normal in all patients with a documented normal AV node function before surgery. One patient had an iatrogenic third degree AV block. There was no difference in PA or HV interval after surgery. Sustained atrial tachyarrhythmias could be induced in 5 patients, of whom 4 developed permanent AF/atrial flutter late after surgery. At late follow-up, (mean 45 months), 27 (73%) patients were in sinus rhythm, 5 (13%) patients had permanent pacing, and 5 patients had recurrent AF requiring His bundle ablation (n = 2) or medical treatment (n = 3). Based on electrophysiological studies, the maze (III) procedure does not cause permanent damage to the sinus or AV nodes or to the right atrial and His-Purkinje conduction systems. Electrophysiological study evaluation may predict the need for postoperative pacemaker. Induction attempts of atrial arrhythmias may predict future recurrences and guide therapeutic efforts. PMID- 14764171 TI - Sleep apnea: a new indication for cardiac pacing? AB - In the general adult population, prevalence of sleep apnea syndrome reaches 4% in men and 2% in women. Continuous positive airway pressure is the most efficient treatment. At the present time, although severe atrial bradycardias could occur during sleep apnea episodes, cardiac pacing has not been demonstrated as an efficient treatment for those bradycardias. Treating sleep apnea generally reduces the number of bradyarrhythmias. However, recent studies reported a beneficial effect of atrial pacing on the sleep apnea burden. The mechanisms rely on two phenomena: first to counteract nocturnal hypervagotonia, and second to treat heart failure. By increasing the heart rate, cardiac output improves, which mitigates pulmonary subedema. Consequently, stimulation of the pulmonary afferent vagal fibers is diminished, which reduces central sleep apnea incidence. During nocturnal hypervagotonia, snoring and obstructive apnea episodes are increased, mainly due to an excessive muscular relaxation of the upper airway area inducing cyclical substantial decreases in the airway caliper. In patients with a low heart rate, atrial pacing can counteract hypervagotonia by enhancing the sympathetic tone and modifying the degree of vigilance. Accordingly, in the near future, sleep apnea treatment might potentially rely on atrial pacing in bradycardic patients with hypervagotonia (with or without heart failure). The role of the physician would then be not only to diagnose sleep apnea, but also to identify potential responders to cardiac pacing. PMID- 14764172 TI - Effects of intravenous nifekalant, a class III antiarrhythmic drug, on atrial vulnerability parameters in patients with paroxysmal atrial fibrillation. AB - Nifekalant, a class III antiarrhythmic drug, has been shown to suppress ventricular tachyarrhythmias, but its effects on AF are unclear. The aim of this study was to clarify the effects of nifekalant on the atrial vulnerability parameters in patients with paroxysmal AF. The study included 18 patients with paroxysmal AF who underwent electrophysiological study before and after intravenous infusion of nifekalant. The atrial electrophysiological parameters including the atrial effective refractory period (AERP), maximum intraatrial conduction delay, and wavelength index, calculated as the ratio of AERP to the maximum conduction delay, were quantitatively measured at baseline and during nifekalant infusion. The mean AERP was significantly prolonged from 214 +/- 27 ms at baseline to 242 +/- 39 ms after nifekalant (P < 0.001). Although earlier studies have shown that nifekalant does not affect the atrial conduction time, the mean maximum conduction delay of the study patients was significantly prolonged from 59 +/- 19 ms at baseline to 72 +/- 28 ms after nifekalant (P = 0.015). There was no significant difference in the wavelength index at baseline (4.1 +/- 1.7) and after nifekalant (4.1 +/- 2.5). However, when the differences of AERP and wavelength index were defined as each parameter during nifekalant infusion minus that at baseline, the difference of AERP showed a direct positive correlation with that of the wavelength index (P = 0.013). In conclusion, nifekalant may be effective in the prevention of AF due to prolongation of the AERP. However, in those patients who have a lesser degree of prolongation of the AERP by nifekalant, the wavelength index tended to be decreased, suggesting that the drug might augment the propensity for AF. PMID- 14764173 TI - Comparison of step-down and binary search algorithms for determination of defibrillation threshold in humans. AB - Determination of DFT is an integral part of ICD implantation. Two commonly used methods of DFT determination, the step-down method and the binary search method, were compared in 44 patients undergoing ICD testing for standard clinical indications. The step-down protocol used an initial shock of 18 J. The binary search method began with a shock energy of 9 J and successive shock energies were increased or decreased depending on the success of the previous shock. The DFT was defined as the lowest energy that successfully terminated ventricular fibrillation. The binary search method has the advantage of requiring a predetermined number of shocks, but some have questioned its accuracy. The study found that (mean) DFT obtained by the step-down method was 8.2 +/- 5.0, whereas by the binary search method DFT was 8.1 +/- 0.7 J, P = NS. DFT differed by no more than one step between methods in 32 (71%) of patients. The number of shocks required to determine DFT by the step-down method was 4.6 +/- 1.4, whereas by definition, the binary search method always required three shocks. In conclusion, the binary search method is preferable because it is of comparable efficacy and requires fewer shocks. PMID- 14764174 TI - Incidence of antitachycardia therapy suspension due to magnet reversion in implantable cardioverter defibrillators. AB - Electromagnetic interference may result in transient or persistent suspension of antitachycardia therapies in ICDs. The incidence of such events has not been assessed so far. Patient charts were retrospectively analyzed for the occurrence of temporary suspension of antitachycardia therapies as it is stored in the Holter of St. Jude Medical or Ventritex ICDs. Follow-up data of 46 patients and 83.7-patient years were analyzed. Overall, 43 episodes of transient ICD inactivation occurred. Twenty-two of these episodes were related to intentional ICD inactivation in the emergency room or during surgery and 12 episodes were related to ICD follow-up. In nine episodes an environmental source of electromagnetic interference is presumed. None of the interactions resulted in persistent ICD inactivation or reprogramming of the devices. The risk for temporary suspension of ICD therapies unrelated to surgery, intentional magnet application in the emergency room, or routine follow-up is 11% per patient and year. Evaluation of its potential sources and the prevalence of ICD inhibition is warranted. PMID- 14764175 TI - Radiofrequency catheter ablation in small children: relationship of complications to application dose. AB - Little data exists to support the use of procedural modifications during radiofrequency catheter ablation (RFCA) in small children. A single institution database was reviewed for patients under 15 kg undergoing RFCA from January 1998 to August 2001. Of 268 RFCA procedures, 18 were done in 14 patients under 15 kg (median weight 5.7 kg, 3.5-13.7; age 5.8 months, 1.2-19.8). Six patients had normal hearts, 4 had congenital heart disease, and 4 patients had cardiomyopathy. Diagnoses were orthodromic reciprocating tachycardia (ORT) in nine patients/nine studies, chaotic atrial tachycardia (CAT) in one patient/two studies, and VT in four patients/seven studies. RFCA variables included maximum temperature (69 degrees C, 50-78), total applications (10, 2-21), applications > 20 seconds (5, 0 15), and total application time (331 s, 26-1,006 s). Complications were pericardial effusion in 1 patient, mild mitral regurgitation in 1, and myocardial infarction in 1 patient. When indexed for weight, the number of applications with a duration > 20 seconds in the ORT group was significantly greater in complicated versus uncomplicated procedures (0.7 applications/kg vs 0.16 applications/kg, P = 0.05). In addition, for the ORT subgroup, the indexed total application time trended higher in complicated versus uncomplicated procedures (40.6 s/kg, vs 6.6 s/kg, P = 0.1). RFCA success was 9/9 in ORT, 6/7 in VT, and 0/2 in CAT. RFCA can be successful in small children; however, complications appear to be related to RF dose indexed for body size. Thus, the decision to proceed with RFCA, and the application duration and number should be guided by patient size, balanced against the risks of the arrhythmia, and reserved for dire circumstances. PMID- 14764176 TI - Lower observed versus expected (based on U.S. age and gender specific rates) survival in patients treated for near-fatal ventricular arrhythmias. AB - Implantable cardioverter defibrillators (ICDs) have improved survival for patients with ventricular fibrillation (VF) or sustained vertricular tachycardia (VT). However, the survival of these patients compared to the general population has not been assessed. Observed survival rates for patients randomized to either antiarrhythmic drug therapy (mainly amiodarone) arm or ICD arm were compared to expected rates, calculated using age and sex-specific survival rates derived from the 1989-1991 US population life tables and applied to the age and sex distribution of patients in each arm. Consistent with the results of the Antiarrhythmics Versus Implantable Defibrillators (AVID) trial, patients randomized to receive ICDs experienced significantly higher survival than those in the drug arm; however, both groups experienced significantly lower survival than expected using age and gender matched U.S. survival rates. Within arms, the difference between the observed and expected rates increased over 3 years of follow-up from 7.7% to 15.3% for the ICD arm, and from 14.6% to 26.4% for the drug arm. These results quantify the improvements in survival that can be expected for VF or VT patients using drug or ICD therapies and underscore the need for continued research into methods for further improving the overall level of health of these patients. PMID- 14764177 TI - The different electrophysiological characteristics in children with Wolff Parkinson-White syndrome between those with and without atrial fibrillation. AB - Atrioventricular reciprocating tachycardia (AVRT) is known to be the most common supraventricular tachycardias in childhood. Because AF with rapid ventricular response may degenerate to ventricular fibrillation through conduction of accessory pathways (APs), it can be potentially life-threatening in some pediatric patients with WPW syndrome. However, information about WPW syndrome children associated with AF is limited. The purpose of this study was to investigate the specific electrophysiological characteristics in pediatric patients with WPW syndrome and AF. From July 1992 to February 2002, 51 pediatric patients with manifest WPW syndrome and documented AVRT underwent electrophysiological study and radiofrequency catheter ablation. In these patients, two (4%) were found to have several spontaneous episodes of AF recognized on 12-lead standard ECG or 24-hour Holter monitoring. Eleven (22%) patients had AF induced by rapid atrial pacing during the baseline procedure of electrophysiological study. The children with manifest WPW syndrome were divided into two groups: those with AF (group 1; n = 11) consisted of seven male and four female children (mean age 15 +/- 3 years, range 10-18), and those without AF (group 2; n = 40) consisted of 22 boys and 18 girls (mean age 16 +/- 3 years, range 7-18). The study excluded a patient who had Ebstein's anomaly associated with moderate tricuspid regurgitation and right atrial enlargement. The onset and duration of symptoms were not significantly different between the two groups. Comparing the electrophysiological characteristics, the atrial effective refractory period (ERP) was shorter in WPW syndrome children with AF (170 +/- 36 vs 190 +/- 38 ms, P = 0.041). This study demonstrated that the pediatric WPW syndrome patients with AF had different electrophysiological characteristics from those without AF. PMID- 14764178 TI - Left atrial vein pacing: a technique of biatrial pacing for the prevention of atrial fibrillation. AB - Biatrial pacing is a promising new therapy for drug refractory AF. This article reports two studies. First, an initial 14-patient experience with a novel technique for biatrial pacing. The authors attempted to pace from the LA vein branches of the proximal CS for LA stimulation. LA vein pacing would potentially offer the advantages of greater interatrial synchronization and possibly greater reduction in AF burden and also of lesser far-field R wave sensing and greater lead stability. Second, a postmortem series examining the number, size, and site of LA veins draining into the proximal CS is described. LA vein pacing was successful in 9 of 14 patients. LA vein electrode parameters have been stable during a median follow-up of 580 days. There were three early lead dislodgments but no other complications. In the second study, a postmortem analysis of 43 human hearts was performed. The study found that 38 (88.4%) of 43 hearts had at least one LA vein draining into the proximal 5 cm of the CS. In addition, 81.2% (33/43) had at least one vein greater than 4 Fr caliber. Thus, pacing in a greater proportion of patients might be achieved by the development and use of smaller (3, 4, and 5 Fr) electrodes. Furthermore, these smaller leads would obviously allow deeper advancement into the LA veins with the potential advantages of greater interatrial synchronization and lead stability and lesser far-field R wave sensing. PMID- 14764179 TI - Alternating narrow and wide complex tachycardia: what is the mechanism? PMID- 14764180 TI - Cyclic appropriate mode switching and inappropriate back switching of a biventricular pacemaker during atrial tachyarrhythmia. AB - In patients with resynchronization devices and intact intrinsic AV conduction, atrial tachyarrhythmias may give rise to high ventricular rates, resulting in inhibition of (bi)ventricular pacing and concomitant lack of therapeutic effects of the device. This report presents a patient with atrial arrhythmias in whom mode switching and back switching of the biventricular pacemaker occurred, due to special timing of the atrial and ventricular deflections. This case report stresses the importance of strenuous treatment of atrial arrhythmias in patients with resynchronization devices. PMID- 14764181 TI - Persistent tachycardia originating from the superior vena cava as a driver for atrial fibrillation. AB - We describe a patient with drug-resistant paroxysmal atrial fibrillation who underwent radiofrequency catheter ablation of tachycardia originating from the superior vena cava (SVC). A continuous and rapid tachycardia conducted to the atrium with evidence of exit block and drove atrial fibrillation (AF) and atrial tachycardia. Neither AF nor atrial tachycardia could be induced after SVC isolation, and nor were they observed during follow-up. We conclude that continuous and rapid tachycardia originating from the SVC can act as a driver as well as an initiator of AF. PMID- 14764182 TI - Atrial standstill with subnodal conduction defect and diffuse ventricular fibrosis managed with low right atrial pacing. AB - In patients with congenital heart disease presenting with bradyarrhythmia, many electrophysiological abnormalities are found. This report presents a case of a man with surgically corrected single atrium and pulmonary valve stenosis, atrial standstill, delayed AV node conduction, first-degree intra- and second-degree infra-His block, who was permanently paced from a restricted excitable area of the low right atrium, as the pacing threshold was unacceptable at any ventricular site. PMID- 14764183 TI - Nonsurgical transthoracic epicardial ablation for the treatment of a resistant posteroseptal accessory pathway. AB - A case of Wolff-Parkinson-White syndrome successfully treated by transcutaneous epicardial radiofrequency ablation is described in a patient with a posteroseptal accessory pathway who had failed prior attempts of conventional endocardial and coronary venous system approaches. Simultaneous endocardial and pericardial space mapping was performed and only ablation from the pericardial space was successful, suggesting an epicardial course of the accessory pathway. PMID- 14764184 TI - Delayed presentation of totally avulsed right superior vena cava after extraction of permanent pacemaker lead. AB - Pacemaker lead extraction has been shown to be an effective and safe treatment for infected permanent pacemaker leads, however, they may lead to potentially serious complications, usually occurring during the extraction procedure. This report describes a case of a 48-year-old woman with a patent persistent left SVC and an infected permanent pacemaker lead of a DDD pacing system who underwent transvenous laser-assisted lead extraction using a combined SVC and femoral approach. Two days after the procedure the patient developed symptoms of SVC obstruction requiring surgical intervention. The right SVC was found to be almost completely destroyed with only a thin strip of the lateral wall intact and active bleeding. The probable causative mechanisms and surgical management are discussed. PMID- 14764185 TI - Dislodgment of an atrial screw-in pacing lead 10 years after implantation. AB - Dislodgment of an atrial screw-in pacing lead is quite rare. This report describes a rare case of an atrial screw-in lead dislodgment 10 years after implantation. Although it is an uncommon complication, very late dislodgment can occur postoperatively, and careful follow-up is necessary. PMID- 14764186 TI - Cardiac surgery for arrhythmias. AB - Cardiac arrhythmia surgery was initiated in 1968 with the first successful division of an accessory AV connection for the Wolff-Parkinson-White Syndrome. Subsequent surgical procedures included the left atrial isolation procedure and the right atrial isolation procedure for automatic atrial tachycardias, discrete cryosurgery of the AV node for AV nodal reentry tachycardia, the atrial transection procedure, corridor procedure and Maze procedure for atrial fibrillation, the right ventricular disconnection procedure for arrhythmogenic right ventricular tachycardia, the encircling endocardial ventriculotomy, subendocardial resection procedure, endocardial cryoablation, the Jatene procedure, and the Dor procedure for ischemic ventricular tachycardia. Because of monumental strides in the treatment of most refractory arrhythmias by endocardial catheter techniques during the past decade, the only remaining viable surgical procedures for cardiac arrhythmias are the Maze procedure for atrial fibrillation and the Dor procedure for ischemic ventricular tachycardia. Nevertheless, the 25 30 years of intense activity in the field of cardiac arrhythmia surgery provided the essential foundation for the development of these catheter techniques and represent one of the most exciting and productive eras in the history of medicine. In one short professional career, we have witnessed the birth of arrhythmia surgery, its adolescence as an "esoteric" specialty, its prime as an enlightening yet exhausting period, and finally its waning years as a source of knowledge and wisdom on which better methods of treatment have been founded. One could hardly ask for a more rewarding experience. PMID- 14764187 TI - Fiscal reality to the practice of medicine. PMID- 14764188 TI - Revisiting the Health Belief Model: nurses applying it to young families and their health promotion needs. AB - The Health Belief Model (HBM) was reviewed with the aim of modifying it so that it reflected a health promotion stance for young families. Since this model's inception, health professionals like nurses have been involved in using the HBM to guide their practice. It is argued that to assist families, nurses now need a model that is focused on "health." In support of this approach, reorienting the HBM and basing it on "positive" health definitions associated with health promotion, by modifying it through adding the constructs "perceived behavioral control" (representing health locus of control) and "behavioral intention" from Ajzen will provide nurses with a more appropriate and useful model for interacting with families and their preschool children. A summary of positive and negative aspects of the modification of the HBM is presented, followed by a strategy for the process of validating the revised HBM for young families. PMID- 14764189 TI - Behavioral and endocrinological evaluation of music therapy for elderly patients with dementia. AB - The present study investigated the effectiveness of music therapy for dementia patients using endocrinological and behavioral evaluations. The study comprised 10 patients with senile dementia who received music therapy; six had Alzheimer's dementia and four had vascular dementia. Music therapy was performed twice a week for 8 consecutive weeks (16 sessions). As a result, total scores on the Mini Mental State Examination (MMSE) did not significantly change, but the scores of a subscale, "language", improved significantly. According to the Multidimensional Observation Scale For Elderly Subjects (MOSES), scores for "irritability" decreased significantly. Regarding changes in salivary chromogranin A (CgA) levels, the average was significantly decreased before session 16 compared to after this. These results suggest that the combination of endocrinological measurements, behavioral evaluations and functional assessment methods are useful in evaluating the effects of music therapy in persons with senile dementia. PMID- 14764190 TI - Swedish mental health nurses' responsibility in supervised community care of persons with long-term mental illness. AB - The aim of the present study was to describe psychiatric nurses' experience of how the changing focus of mental health care in Sweden, from in-patient treatment to community-based care, has influenced their professional autonomy. Eleven psychiatric nurses were interviewed and a qualitative content analysis was used to identify major themes in the data. Three main themes were found: pattern of responsibility, pattern of clinical judgement, and pattern of control through support and supervision. All themes were related to the nurse's identity, moral responsibility and the feelings of loneliness and independence in his/her daily work. Together, the three themes were found to constitute a process. This study shows the complexity involved in nursing care provided in the patient's home. Achieving control over the patient's everyday life through support and supervision does not imply taking over the patient's autonomy, but rather reducing the stigma attached to mental illness and facilitating the process of rehabilitation. PMID- 14764191 TI - Information needs and the related variables of Japanese family caregivers of terminally ill cancer patients. AB - The present study examined the information needs of family caregivers of terminally ill cancer patients. Data were obtained by a structured interview from 66 Japanese caregivers of cancer patients institutionalized in a palliative care unit. Needs for disease-related information (the disease, treatment and prognosis) and care-related information (the care for the patient and family members) were investigated, and demographic and situational characteristics related to individual information needs were assessed. More than 60% of family caregivers wanted disease-related information, and approximately half of these wanted patient and family care-related information. These information needs had significant correlations with the family caregiver's age and with such patient dependent situational variables as time since diagnosis, care site before enrolling in a palliative care unit, and presence or absence of other family caregivers. The results may help health care professionals to better inform terminally ill cancer patients and their family caregivers. PMID- 14764192 TI - Measuring the impact of housing on people with schizophrenia. AB - Review of the available literature, from various countries, on housing options indicates that, for people with a mental illness, boarding houses are the least desirable type of community accommodation and that living in their own home is the most desirable type of accommodation. The present research project provides a more in-depth examination of people with schizophrenia and the impact of living in their own home compared to living in a boarding house. In this Australian study there were 3231 subjects, 3033 who were living in their own homes and 201 living in boarding house accommodation. The study used two instruments from the Mental Health Classification and Service Cost Project, specifically the Health of the Nation Outcomes Scale, which is a measure of current symptoms, and a shortened version of the Life Skills Profile, which measures global level of functioning. Results indicated that while there were no differences in the level of psychiatric symptoms experienced, people living in boarding houses had less access to social support, meaningful activities and work; they also had a significantly lower level of global functioning. These findings contradict the conventional wisdom that people with schizophrenia resort to living in boarding houses because of their level of disability and highlights an area of potential intervention for community health services. PMID- 14764193 TI - Effects of nurses' sex-role orientation on positive and negative assertion. AB - The present paper examines how the sex-role orientation of the nurse can affect both positive and negative assertion. The Bem Sex-Role Inventory (BRSI) and the Assertion Inventory (AI) were used to collect self-reported data from a convenience sample of 94 subjects. Behavioral data were obtained by directly sampling 50 nurses' responses within role-play situations. Contrary to earlier studies, the findings of the investigation indicated that there was no significant relationship between assertion measurements and sex-role orientation. Following a discussion of the findings, recommendations relating to future research are made and the limitations of the study are identified. PMID- 14764194 TI - Eating behaviors of elderly persons with hyperlipidemia in urban Chiang Mai. AB - The objective of the present ethnographic study was to describe the eating behaviors of elderly persons with hyperlipidemia. Twenty-one elderly subjects with hyperlipidemia and their relatives volunteered to be included in the study. Focus group and in-depth interviews with participant observations were used for data collection. Each transcript from interviews and observations were analyzed using content analysis. Subjects were asked to validate findings and ethical considerations were maintained throughout the study. When discussing the meaning of food, three themes emerged: (i) substances make people strong or healthy; (ii) food is like medicine and; (iii) food causes illness. Most subjects had learned about food and nutrition from their parents or relatives and from school. Regarding food procurement, most bought food such as fish or other seafood, meat and eggs from the fresh markets nearby almost daily. All subjects cooked their own traditional northern Thai food, such as Hung-le (northern curry) and preferred seasoning such as fish sauce, pepper and herbs. They ate when they were hungry or after they had finished working. The present study reflected that northern Thai culture strongly influences the participants' eating behaviors. PMID- 14764195 TI - Predictors of nurses' intent to stay at work in a university health center. AB - The purpose of the present study was to evaluate nurses' intent to stay at work and to determine the associations between intent to stay and various predictors. A sample of 108 nurses at a single tertiary care hospital filled in a questionnaire on intent to stay, satisfaction at work, satisfaction with administration, organizational commitment and work group cohesion. The results showed that the majority of nurses are planning to stay in their current job. Satisfaction at work and satisfaction with administration are the best predictors of intent to stay and explained 25.5% of intent to stay variance. It is possible that developing strategies based on the predictors of intent to stay at work could improve that intent. Healthcare organizations could consider this with the objective of increasing nurses' intent to stay at work and, consequently, retention. PMID- 14764198 TI - Message from the editors: consolidation. PMID- 14764199 TI - Mice and methods. PMID- 14764201 TI - Neural regulation of tone in the oesophageal body: in vivo barostat assessment of volume-pressure relationships in the feline oesophagus. AB - Recent combined manometric-barostat studies demonstrated that the oesophageal body exhibits both peristaltic contractions and tone. This study further characterized the neural modulation of tone in the feline oesophageal body. Simultaneous oesophageal barostat and manometry were performed in 20 adult cats under ketamine sedation. Oesophageal tone and peristalsis were assessed in the distal smooth muscle oesophagus. Cholinergic modulation was studied using neostigmine, erythromycin, atropine and vagotomy. Nitrergic regulation was assessed using sildenafil to increase cellular cyclic guanosine monophosphate and the nitric oxide synthase blocker Nomega-nitro-l-arginine (l-NNA). The presence of a tonic contractile activity in the distal oesophageal body was confirmed. Peristaltic contractions proceeded along the oesophageal body over the background tonic contraction. Neostigmine and erythromycin enhanced (20-30%) whereas bilateral vagotomy and atropine strongly decreased oesophageal tone (50-60%). However, l-NNA increased (40%) and sildenafil decreased oesophageal tone (30%). Therefore, tonic contractile activity in the oesophageal body is mainly caused by a continuous cholinergic excitatory input. A nitric oxide inhibitory mechanism may have a complementary role in the regulation of oesophageal tone. PMID- 14764200 TI - Central vagal stimulation evokes gastric volume changes in mice: a novel technique using a miniaturized barostat. AB - We have developed a novel technique to measure gastric volume in vivo in mice; this will be invaluable for revealing gastric alterations in genetically modified mice models, thus expanding our understanding of the mechanisms underlying functional disorders. Experimental data on gastric tone currently available has focused on rats using isovolumetric techniques to measure pressure changes, whereas clinical studies use barostatic techniques to measure volume changes. For better translational approaches, we assessed the feasibility of using a miniaturized barostat to measure gastric volume changes in urethane-anaesthetized and unanaesthetized-decerebrate mice. Additionally, we assessed whether central vagal stimulation alters gastric volume in urethane-anaesthetized mice. Nitric oxide donor sodium nitroprusside (1mg kg-1 i.p.) increased gastric volume (+134 +/- 20 microL), whereas the cholinergic agonist carbachol (3 microg kg-1 i.p.) decreased gastric volume (-153 +/- 20 microL). Similar responses were obtained in urethane-anaesthetized and unanaesthetized-decerebrate animals. Microinjection of L-glutamate (25 nmol) into dorsal motor nucleus of the vagus (DMV) altered gastric volume; microinjection into rostral DMV led to gastric contraction (-83 +/- 18 microL) while stimulation of caudal DMV resulted in gastric relaxation (+95 +/- 16 microL). This reveals a functional organization of DMV in mice. This study validates barostatic techniques for application to mice. An understanding of gastric contractility and tone is clinically relevant as impaired gastric accommodation reflex may be an underlying cause of functional dyspepsia. PMID- 14764202 TI - Effect of interval between swallows on oesophageal pressures and bolus movement in normal subjects - Studies with combined multichannel intraluminal impedance and oesophageal manometry. AB - The effect of closely spaced swallows to decrease peristalsis ('deglutitive inhibition') is believed to be due to both central inhibitory impulses and smooth muscle refractoriness. Ten volunteers (three females, age 26-65) were given both four pairs and two series of four swallows at 5-, 10-, 15-s intervals and control swallows at 30-s intervals. Oesophageal function was assessed using combined multichannel intraluminal impedance and oesophageal manometry (MII-OM). Swallows were considered manometrical effective if distal oesophageal pressures >/=30 mmHg. Complete bolus transit was defined as bolus exiting from all three distal impedance segments. During swallowing at 5-s intervals the majority of initial swallows were ineffective with incomplete bolus transit while the last swallow in both series and pairs was manometrically effective with complete bolus transit. During swallowing at 10-15-s intervals the number of manometric ineffective swallows and swallows with incomplete bolus transit progressively increased with the number of swallows. The functional information obtained by MII-OM indicates pooling of liquid in the distal oesophagus that is cleared by the last swallow determined by, previously reported, neural inhibition occurring during swallowing spaced 5 s apart whereas incomplete bolus transit is related to manometrically ineffective swallows resulting from muscle refractoriness occurring during swallowing at 10-15-s intervals. PMID- 14764203 TI - Regional differences in oesophageal motor function. AB - We tested the hypotheses that oesophageal bolus transit and motor function vary regionally, with bolus viscosity and with body position. In healthy volunteers, we measured the bolus head advance time, bolus presence time and bolus transit time in the proximal and distal oesophagus using water and viscous materials. We compared concurrent manometric responses. Bolus head advance time, bolus presence time and bolus transit time were longer in the distal oesophagus during water and viscous swallows in the upright and supine positions. The total bolus head advance time and transit time, measured across the entire oesophageal body, were shorter for water than viscous swallows. The amplitudes of peristaltic pressure waves were lower for viscous swallows, and varied as a function of region. These studies demonstrated true functional differences between the proximal and distal oesophagus using multichannel intraluminal impedance and that the viscosity of the bolus is a determinant of oesophageal function. PMID- 14764204 TI - Isolation and characterization of resident macrophages from the smooth muscle layers of murine small intestine. AB - Macrophages within the murine tunica muscularis were isolated and cultured for physiological studies. Following dispersion, macrophages were identified by phagocytotic activity of fluorescein isothiocyanate (FITC)-dextran. Immediately following isolation, macrophages were rounded and possessed fluorescent granula but developed a ramified shape after 3-4 days in culture. Resident and cultured macrophages were immunopositive for F4/80 and I-Ad/I-Ed. Greater than 90% of F4/80 positive cultured cells were FITC-dextran positive. Macrophages had resting membrane potentials (RMP) of -33.3 +/- 1.5 mV after 1 day in culture, which increased to -53.9 +/- 4.4 mV after 3-4 days. The change in RMP was associated with the development of an inward rectifying K+ current, and a decrease in a voltage-dependent, inactivating outward current. After 3-4 days in culture the inflammatory mediated substances adenosine triphosphate (ATP), platelet activating factor and bacterial lipopolysaccharide induced increases in cytoplasmic Ca2+ ([Ca2+]i). Forskolin suppressed the ATP-induced increase in [Ca2+]i. Macrophages exhibited oxidative bursts, measured by oxidation of dihydrorhodamine-123 to rhodamine-123. Oxidative bursts coincided with a reduction in intracellular pH. Macrophages expressed a proton conductance that may participate in pH maintenance during reactive oxygen production. These results suggest that resident macrophages in the intestine may play a role in the immunological protection of the gut. PMID- 14764205 TI - Effects of the probiotic yeast Saccharomyces boulardii on the neurochemistry of myenteric neurones in pig jejunum. AB - We studied the effects of food supplementation with Saccharomyces boulardii (S. boulardii; synonym S. cerevisiae HANSEN CBS 5926; 1 g per day for 9 days) on the presence and co-localization patterns of neuronal markers in myenteric neurones of the pig jejunum. The pan neuronal marker Hu revealed no change in the number of neuronal cell bodies per ganglion (37 +/- 7 in control vs 34 +/- 9 in the S. boulardii group). Ranked by size the following cell populations were identified: choline acetyltransferase (ChAT), calbindin-28k (CALB), substance P (SP), neurofilament 160 kD (NF-160), vasoactive intestinal polypeptide (VIP), nitric oxide synthase (NOS), calcitonin gene-related peptide (CGRP), calretinin (CALRET). We found a significant decrease in the number of CALB myenteric neurones in animals which received S. boulardii supplemented diet. None of the other neuronal markers revealed any difference between controls and S. boulardii treated animals. The study reports transmitter-localization patterns in the myenteric plexus of the pig jejunum and provides evidence that changes in the neurochemistry of enteric neurones occur with S. boulardii supplemented diet. Although only CALB expression was altered and the functional significance of this finding remains unknown, our study identified a possible new effector level of probiotics in the gut. PMID- 14764206 TI - The expression and role of Fas ligand in intestinal inflammation. AB - Fas ligand (FasL) is involved in the pathogenesis of inflammatory diseases and immune privilege. We examined the expression of FasL in the enteric nervous system (ENS) in murine colitis and guinea-pig ileitis. We studied FasL immunoreactivity, functional integrity of the ENS, severity of colitis, and distribution of neutrophils in wild type and B6/gld mice that lack functional FasL. In ileitis, the distribution of FasL, CD4+ and CD8+ T cells was examined. FasL expression was increased in the ENS of wild type mice with colitis, but decreased labelling of nerve fibres was noted in B6/gld mice. Neutrophils were more abundant and widely distributed in B6/gld mice. Colitis was more severe and persistent in B6/gld mice 7 days after induction. Functional parameters of intestinal secretion and motility in B6/gld mice were the same as controls. In ileitis, FasL expression was increased in the guinea-pig ENS and returned to control levels following the resolution of inflammation. While T cells were not present in the ENS of controls, they were observed during inflammation, but were excluded from ganglia. The number of enteric neurons was unchanged over the course of inflammation. The expression of FasL is altered in intestinal inflammation and contributes to its resolution in experimental colitis. PMID- 14764207 TI - Coupling among interstitial cells of Cajal in the human ileum. AB - Current knowledge on the morphology and physiology of interstitial cells of Cajal (ICC) is mostly based on animal studies, and information about the function of these cells in humans is scarce. There is ultrastructural evidence that ICC in the myenteric region (ICC-MP) of the small intestine of several species are connected by gap junctions, but these were not observed in the human small intestine. The aim of the present study was to determine whether functional coupling also exists among ICC-MP in the human ileum. We visualized ICC-MP in live tissues using Nomarski optics, and verified their identity by staining for c Kit. ICC were injected intracellularly with the fluorescent dye Lucifer yellow, which crosses gap junctions. In most cases the labelled cells had oval somata with two primary processes. At normal pH (7.3-7.4) only 20.2% (21/104) of the injected ICC were coupled to other ICC. However, at pH 7.8-7.9 coupling incidence increased to 74.5% (35/47, P < 0.0001). The injected cells were coupled to one to 35 other ICC. Octanol blocked coupling in all cases. Apparently, gap junctions interconnect ICC in the human small intestine. Coupling was enhanced by a small increase in pH, suggesting that it may be under physiological control. PMID- 14764208 TI - Nitrergic-purinergic interactions in rat distal colon motility. AB - Responses of rat distal colon circular muscle strips to exogenous nitric oxide (NO) and adenosine 5'-triphosphate (ATP) and to electrical field stimulation (EFS) were assessed in the absence/presence of various agents that interfere with nitrergic-purinergic pathways. Exogenous NO (10-6 to 10-4 mol L-1) elicited concentration-dependent, tetrodotoxin (TTX)-insensitive relaxations. The soluble guanylyl-cyclase (sGC) inhibitor 1H[1,2,4,]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ) reduced duration and amplitude; the small conductance Ca2+-sensitive K+ (SK)-channel blocker apamin (APA) only shortened the relaxations. ODQ + APA showed a marked inhibitory effect on duration and amplitude. TTX, APA, the NO synthase inhibitor N(omega)-nitro-l-arginine methyl ester (l-NAME) and the purinergic receptor P2Y antagonist Reactive Blue 2 (RB2) shortened the relaxations by exogenous ATP (10-3 mol L-1) but did not influence the amplitude. ODQ had no effect. TTX + l-NAME did not yield a more pronounced inhibitory effect than TTX alone. The effect of ATP-gamma-S was similar to that of ATP. Electrical field stimulation (EFS) (40 V, 0.05 ms, 0.5-4 Hz for 30 s) yielded TTX-sensitive relaxations that were not altered by l-NAME, ODQ or RB2. APA shortened the relaxations. l-NAME + APA nearly abolished these relaxations. ODQ + APA and RB2 +l-NAME reduced the duration. These results suggest that distinct sets of small conductance SK-channels are involved in the amplitude and the duration of the relaxations and that NO increases their sensitivity to NO and ATP via guanosine 3',5'-cyclic monophosphate (cGMP). ATP elicits relaxations via P2Y receptors with subsequent activation of SK-channels and induces neuronal release of NO. Both nitrergic and purinergic pathways must be blocked to inhibit EFS-induced relaxations. PMID- 14764209 TI - New probe for the measurement of dynamic changes in the rectum. AB - Conventional mano-volumetric techniques cannot measure changes in circumferential dimensions at several axial positions within a bowel segment. Our aims were to validate a new impedance planimetry probe for simultaneously measuring the cross sectional area (CSA) at five axial positions in vitro and in vivo in 10 anesthetized pigs. The day-to-day coefficient of variation (CV) for CSA measured by the probe in cylindrical tubes of known diameter was 0.8-9.5%. The mean from actual diameter deviation ranged from 2.3 to 6.7%. In a conical tube the day-to day CV was 2.3-8% and mean percentage deviation -2.8 to 1.0. Interposed narrowing sections caused a total CV of 7-13%. In vivo studies revealed variations in CSA, associated with expulsion of flatus. It is concluded that impedance planimetry allows simultaneous measurement of CSA at several levels within the rectum. In vitro validity was acceptable and alterations in lumen diameter were identified in vivo. PMID- 14764210 TI - Influence of continuous isobaric rectal distension on gastric emptying and small bowel transit in young healthy women. AB - Patients with slow transit constipation frequently have delayed gastric emptying. In animals rectal distensions inhibit gastrointestinal motility. In healthy volunteers isovolumetric rectal distensions delay upper gut transit. The purpose of this study was to determine the effect of continuous isobaric rectal distension on gastric emptying and oro-cecal transit in young females. Using validated 13C octanoic and lactose-[13C] ureide breath tests gastric half emptying time and oro-cecal transit time for a meal were measured in 12 volunteers. The tests were repeated in randomized order: during isobaric balloon distension and during sham distension. Isobaric rectal distension was applied using a polyethylene bag connected to a barostat. Intraballoon pressure was kept just below the threshold for the urge sensation. Mean gastric half-emptying time during rectal distension (92.3 +/-5.1 min) was significantly higher than during sham distension (78.8 +/- 4 min; P = 0.015). Mean oro-cecal transit time during rectal distension (391.3 +/-29.1 min) and sham distension (328.8 +/- 38.4 min) were not significantly different. In conclusion, these findings indicate that isobaric rectal distension inhibits gastric emptying, but not small bowel transit in young healthy women. Studies in patients with constipation are indicated. PMID- 14764211 TI - Characterization of the primary spinal afferent innervation of the mouse colon using retrograde labelling. AB - Visceral pain is the most common form of pain produced by disease and is thus of interest in the study of gastrointestinal (GI) complaints such as irritable bowel syndrome, in which sensory signals perceived as GI pain travel in extrinsic afferent neurones with cell bodies in the dorsal root ganglia (DRG). The DRG from which the primary spinal afferent innervation of the mouse descending colon arises are not well defined. This study has combined retrograde labelling and immunohistochemistry to identify and characterize these neurones. Small to medium sized retrogradely labelled cell bodies were found in the DRG at levels T8-L1 and L6-S1. Calcitonin gene-related peptide (CGRP)- and P2X3-like immunoreactivity (LI) was seen in 81 and 32%, respectively, of retrogradely labelled cells, and 20% bound the Griffonia simplicifolia-derived isolectin IB4. CGRP-LI and IB4 were co-localized in 22% of retrogradely labelled cells, whilst P2X3-LI and IB4 were co-localized in 7% (vs 34% seen in the whole DRG population). Eighty-two per cent of retrogradely labelled cells exhibited vanilloid receptor 1-like immunoreactivity (VR1-LI). These data suggest that mouse colonic spinal primary afferent neurones are mostly peptidergic CGRP-containing, VR1-LI, C fibre afferents. In contrast to the general DRG population, a subset of neurones exist that are P2X3 receptor-LI but do not bind IB4. PMID- 14764212 TI - Endogenous endothelin increases gallbladder tone and leads to acute cholecystitis in the Australian possum. AB - Endothelins are bioactive peptides produced by gallbladder epithelial cells. We aimed to determine the role of endothelins in acute cholecystitis. Escherichia coli lipopolysaccharide vs saline (sham) was instilled into the gallbladder lumen of Australian possums. Some animals received the non-selective endothelin antagonist, tezosentan. At 4 or 24 h, plasma and gallbladder endothelins and white blood cell count (WBCC) were determined. Acute cholecystitis was assessed using a histopathology score. In other animals gallbladder tone was determined. At 4h, a dose-dependent 60-fold increase in gallbladder endothelin level occurred (P = 0.001) but other parameters remained comparable with sham animals. Epithelial cells were endothelin-immunoreactive. At 24 h, the WBCC rose (P < 0.007), and severe cholecystitis developed. Gallbladder but not plasma endothelin levels remained elevated. Tezosentan pre-treatment resulted in a histologically normal gallbladder, but the WBCC and gallbladder endothelin levels were elevated. Lipopolysaccharide or saline instillation also caused a time-dependent increase in gallbladder tone over 4 h (P < 0.001), but not in control animals. This increase was reduced by tezosentan treatment. Gallbladder endothelin production is an early event in acute cholecystitis, increases gallbladder tone and plays a crucial role in the inflammatory process. PMID- 14764213 TI - Noninvasive monitoring of deterioration in skeletal muscle function with forearm cast immobilization and the prevention of deterioration. AB - BACKGROUND: In this research inactivity was simulated by immobilizing the forearm region in a plaster cast. Changes in skeletal muscle oxidative function were measured using near-infrared spectroscopy (NIRS), and the preventative effect of the training protocol on deterioration of skeletal muscle and the clinical utility of NIRS were examined. METHODS: Fourteen healthy adult men underwent immobilization of the forearm of the non-dominant arm by plaster cast for 21 days. Eight healthy adult subjects were designated as the immobilization group (IMM) and six were designated as the immobilization + training group (IMM+TRN). Grip strength, forearm circumference and dynamic handgrip exercise endurance were measured before and after the 21-day immobilization period. Using NIRS, changes in oxidative function of skeletal muscles were also evaluated. Muscle oxygen consumption recovery was recorded after the completion of 60 seconds of 40% maximum voluntary contraction (MVC) dynamic handgrip exercise 1 repetition per 4 seconds and the recovery time constant (TcVO2mus) was calculated. RESULTS: TcVO2mus for the IMM was 59.7 +/- 5.5 seconds (average +/- standard error) before immobilization and lengthened significantly to 70.4 +/- 5.4 seconds after immobilization (p < 0.05). For the IMM+TRN, TcVO2mus was 78.3 +/- 6.2 seconds before immobilization and training and shortened significantly to 63.1 +/- 5.6 seconds after immobilization and training (p < 0.05). CONCLUSIONS: The training program used in this experiment was effective in preventing declines in muscle oxidative function and endurance due to immobilization. The experimental results suggest that non-invasive monitoring of skeletal muscle function by NIRS would be possible in a clinical setting. PMID- 14764215 TI - [Neural factors and their effects on wound healing]. PMID- 14764214 TI - Estimation of the time-course of dopamine D2 receptor occupancy in living human brain from plasma pharmacokinetics of antipsychotics. AB - Although the kinetic profile of antipsychotics at dopamine D2 receptor sites has been suggested to be important for antipsychotic action and dosing schedule, the kinetic profiles of the respective antipsychotic drugs in the brain have not yet been clearly defined. We aimed to estimate the time-course of dopamine D2 receptor occupancy from plasma pharmacokinetics and the apparent in-vivo affinity parameter (ED50; concentration required to induce 50% occupancy). Dopamine D2 receptor occupancies and plasma concentrations of risperidone were measured in five patients with schizophrenia using positron emission tomography with [11C]FLB 457. Measured dopamine D2 occupancies were compared with those estimated from plasma kinetics and in-vivo ED50. The time-course of dopamine D2 receptor occupancy was simulated with altered plasma kinetics or apparent in-vivo affinity parameters of the drug. Mean half-life of dopamine D2 receptor occupancy of risperidone was 80.2 h while that of the plasma concentration was 17.8 h. Dopamine D2 receptor occupancy estimated from plasma pharmacokinetics and in-vivo ED50 was within 1 S.D. of the mean measured occupancy. When the ED50 value was changed to one-tenth and 10-fold, the simulated half-life of receptor occupancy changed to 117.6 h and 27.3 h respectively. Using plasma pharmacokinetics and in vivo ED50, the time-course of receptor occupancy could be calculated. Simulation of drug kinetics at receptors would provide useful information for the evaluation of antipsychotics. PMID- 14764216 TI - [A novel strategy of immno-modulation in sepsis]. PMID- 14764217 TI - [Dynamic change in interleukin-18 and its relationships with multiple organ dysfunction syndrome]. AB - OBJECTIVE: To determine interleukin-18 (IL-18) in plasma after trauma, and evaluate its relationship with multiple organ dysfunction syndromes. METHODS: A total of 54 patients were enrolled and divided into low injury seriousness score (L-ISS) and high injury seriousness score (H-ISS) groups. Blood IL-18 levels were determined on admission, day 4, day 7 and day 14 after trauma using enzyme-linked immunoadsorbent assay (ELISA), and the relationship between its level and organ dysfunction was analyzed. RESULTS: Plasma IL-18 concentration was significantly increased on posttrauma day 4 and day 7. Also, plasma IL-18 concentration was significantly higher in patients with organ dysfunction. CONCLUSION: This study suggests that major trauma can lead to a marked elevation of IL-18, which might be associated with organ dysfunction. PMID- 14764218 TI - [Gene expression of toll-like receptors in the liver, lungs and spleen in mice after endotoxin challenge]. AB - OBJECTIVE: To investigate the gene expression of some lipopolysaccharide (LPS) receptors after LPS stimulation. METHODS: The total RNA from normal and LPS challenged mice was extracted by Trizol reagent and the gene expression of Toll like receptor 2 (TLR2), TLR4, CD(14), LPS-binding protein (LBP) and tumor necrosis factor-alpha(TNF-alpha) were measured by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR). RESULTS: The gene of TLR2 was expressed in normal lungs and spleen tissues, and TLR2, CD(14), LBP in liver. After challenged by LPS, the expressions of TLR2, TLR4, and TNF-alpha in lungs, TLR2, CD(14), LBP and TNF-alpha in liver, TLR2, TLR4, CD(14), and TNF-alpha in spleen were increased at 1, 3, and 5 hours. CONCLUSION: LPS might alert the ability against pathogen-associated molecules by inducing or enhancing the expression of genes that involved in the LPS signal transduction. PMID- 14764219 TI - [Experimental study on composite transplantation of acellular porcine dermal matrix and micro-autograft to repair deep burn wound]. AB - OBJECTIVE: To observe the effect of composite skin graft consisting of heterogeneous (swine) acellular dermal matrix and combined micro-autograft and allograft with the aim at overcoming shortcomings of repairing materials for burn wound. METHODS: Forty-eight Sprague-Dawley rats with full-thickness skin defect on the back were grafted with the composite skin (compound skin grafting group, G) and autologous particulate skin (control group, C), respectively. The area of wound healing and the rates of wound contraction were measured. Tissue specimens were harvested for histological examination. RESULTS: The composite skin graft resulted in less contracture and better elasticity. Compared with group C, the wound contraction rate was lower significantly. Histological examination revealed that there was full differentiation of epithelium, orderly collagen arrangement and intact basement membrane. CONCLUSION: Coverage of wound with composite skin composing heterogeneous (swine) acellular dermal matrix and autologous particulate skin could promote wound healing and improve wound healing quality. PMID- 14764220 TI - [Effects of high-volume hemofiltration on serum levels of tumor necrosis factor and its receptors in patients with multiple organ dysfunction syndromes]. AB - OBJECTIVE: To investigate whether serum tumor necrosis factor-alpha (TNF-alpha) and its receptors can be removed by high-volume hemofiltration (HVHF) or continuous veno-venous hemofiltration (CVVH). METHODS: The study was performed in 12 multiple organ dysfunstion syndrome (MODS) patients with acute renal failure (ARF). They were randomized to receive either CVVH (n=10) or HVHF (n=8). TNF alpha and soluble tumor necrosis factor-receptor (sTNF-R1 and sTNF-R2) concentrations were measured in serum by enzyme-linked immunoadsorbent assay (ELISA). RESULTS: Compared with that before the therapy, the average concentrations of plasma creatinine and urea were decreased significantly 8 hours after HVHF or CVVH in MODS patients with ARF (P<0.001). In patients on HVHF, the serum TNF-alpha concentrations were significantly lower 8 hours after treatment (P<0.01) compared with that before treatment, 1 hour and 4 hours after treatment. There were not significant changes in the serum TNF-alpha concentrations in patients on CVVH and the serum sTNF-R1 and sTNF-R2 concentrations in patients on CVVH or HVHF. CONCLUSION: In MODS patients with ARF undergoing HVHF, the serum TNF-alpha concentrations dropped significantly, but the serum sTNF-R1 and sTNF-R2 concentrations do not change significantly. Our study suggests that HVHF may be the better option for the treatment of MODS patients. PMID- 14764221 TI - [Gene expression of angiogenesis-related factors in fetal skin at different developmental stages and childhood skin]. AB - OBJECTIVE: To explore the change in gene expression of angiogenesis-related factors vascular endothelial growth factor (VEGF), angiopoietin-1 (Ang-1), acid fibroblast growth factor (aFGF) and basic fibroblast growth factor (bFGF), in fetal skin at different developmental stages and children skin and their potential biological significances. METHODS: Fetal skin samples of human embryo were obtained from spontaneous abortion at different gestational ages ranging from 13 to 32 weeks, and children skin specimens were collected from child patients (4-12 years) undergoing plastic surgery. After morphological characteristics of skin at different developmental stages were defined histologically gene expressions of VEGF, Ang-1, aFGF and bFGF were examined with reverse transcription-polymerase chain reaction analysis (RT-PCR). RESULTS: The trend of changes in gene expression of VEGF, Ang-1, aFGF and bFGF was not same for different skin specimens at various developmental stages. In early gestational fetal skin, genes of VEGF and Ang-1 were strongly expressed, while in late gestational and childhood skins, gene expressions of VEGF and Ang-1 were apparently decreased. In skin of middle gestational stage, the level of aFGF gene expression was highest, and then it was progressively reduced. In childhood skin, this gene was weakly expressed. In marked contrast, the contents of transcripts of bFGF showed no substantial change in fetal skin at different developmental stages, whilst the mRNA content of bFGF was significantly decreased in childhood skin. CONCLUSION: VEGF, Ang-1, aFGF and bFGF might be involved in regulating angiogenesis in skin from fetuses of different gestational stages and children. The relative increase in gene transcription of VEGF and Ang-1 in younger fetal skin might be one of the reasons why cutaneous cells proliferate rapidly and the wounds heal without scar. PMID- 14764222 TI - [Effects of carbachol injection in intestine on plasma levels of inflammatory medium in rats during gut ischemia/reperfusion]. AB - OBJECTIVE: To investigate the effects of carbachol injection in intestine on plasma levels of tumor necrosis factor-alpha(TNF-alpha), interleukin-10 (IL-10) and cortisol in rats during gut ischemia/reperfusion. METHODS: Wistar rats were anaesthetized with soluble pentobarbitone, and subjected to superior mesenteric artery occlusion (SMAO) for 60 minutes, followed by reperfusion for 60 minutes. Animals were divided into three groups, pretreated group (carbachol injection in intestine at 30 minutes after SMAO; 0.1 mg/kg), treated group (carbachol injection at 30 minutes from onset of reperfusion), and controls (saline injection). Plasma TNF-alpha, IL-10 and cortisol levels were determined at 1.0, 2.5 and 6.0 hours after SMAO. RESULTS: The plasma levels of TNF-alpha significantly decreased in pretreated and treated groups than those in controls after carbachol injection (both P<0.01). However, the levels of IL-10 and cortisol didn't show significant differences among three groups. CONCLUSION: The RESULTS suggest that carbachol can reduce the proinflammatory cytokine release and have a less inhibitory effect on the anti-inflammatory cytokine. It is indicated that carbachol play a potential role in alleviating systemic inflammatory response during splanchnic ischemia/reperfusion injury. PMID- 14764223 TI - [Effects of enteral supplementation with glutamine on mitochondria respiratory function of intestinal epithelium in burned rats]. AB - OBJECTIVE: To investigate the effects of enteral supplementation with glutamine on mitochondria respiratory function of intestinal epithelium in burned rats. METHODS: Wistar rats inflicted with 30% total body surface area (TBSA) full thickness thermal injury were randomly divided into three groups, i.e. burn with enteral nutrition (EN), burn with glutamine treatment (GLN), and normal control (C) groups. Burned rats were infused 732.2 kJ.kg-1.d-1 solution for intravenous nutrition and oral administration, in which the supply energy ratio of glucose, fat and protein was 55:30:15 respectively, glucose was 15.3% and the proportion of calorie to nitrogen was 183:1. The following indices including respiratory control rate (RCR), oxygen extraction (Oext), P/O ratio and intestine mucosal blood flow (IMBF) were measured on postburn days 1, 3, 5, 7, 10. RESULTS: After burn injury, the RCR, Oext, P/O ratio, and IMBF were significant decreased in both EN and GLN groups, but all above indices were markedly increased in GLN group compared to those in EN group. CONCLUSION: After burn injury, the IMBF and Oext were declined, resulting in mitochondria respiratory oxidative dysfunction and phosphorylation discoupling in intestinal epithelium. GLN supplementation appears to be beneficial to improving IMBF, increasing Oext, abating the extent of mitochondria respiration dysfunction, and promoting oxidative phosphorylation. PMID- 14764225 TI - [Study on apoptosis of lung tissue cells in patients with severe brain injury complicated with aspiration-induced lung injury]. AB - OBJECTIVE: To investigate the role of apoptosis of pulmonary cells in aspiration induced lung injury in patients with severe brain injury with or without aspiration-induced lung injury. METHODS: The Glasgow scale (GCS) of 11 dead patients with severe closed brain injury was 3-8. There 11 cases were divided into aspiration-induced lung injury (AILI) and non-aspiration-induced lung injury (NAILI) groups. The plasma levels of tumor necrosis factor-alpha (TNF-alpha) and interleukin-8 (IL-8) were measured, and the ratio of apoptosis in lung tissue cells was also determined. RESULTS: The plasma levels of TNF-alpha and IL-8 in NAILI and AILI groups were (2.17+/-0.41)nug/L vs. (3.14+/-0.28)nug/L and (0.42+/ 0.05)nug/L vs. (0.91+/-0.08) nug/L (P<0.05) respectively. Lung tissue cell apoptosis ratio was significantly higher in AILI group than NAILI group (P<0.01). CONCLUSION: TNF-alpha and IL-8 may induce apoptosis in lung tissues through different signaling pathway. During the early phase of aspiration-induced lung injury complicating severe closed brain injury, apoptosis in cells of lung tissue may play a role in the pathogenes. PMID- 14764226 TI - [Impact factors and clinical significance of the lung injury complicating acute pancreatitis]. AB - OBJECTIVE: To study the impact factors and clinical significance of the acute lung injury (ALI) complicating acute pancreatitis. METHODS: Two hundred and fifty patients with acute pancreatitis were retrospectively analyzed to study the impact of the clinical types, different stages of the disease, systemic inflammatory response syndrome (SIRS) and its duration on the incidence of ALI. Also, the relationship between ALI and severity of the disease, as shown by acute physiology and chronic health evaluation II (APACHE II) score, the incidence of acute respiratory distress syndrome (ARDS) and multiple organ dysfunction syndromes (MODS), and prognosis was analyzed in patients with acute pancreatitis. RESULTS: Our data showed that the clinical types and different stages of acute pancreatitis were significantly related to the incidence of ALI (P<0.01). The incidence of ALI was significantly higher in patients with SIRS compared with those no SIRS (P<0.01), and the longer the duration of SIRS, the higher was the incidence of ALI. In patients with ALI, APACHE II score and incidence of ARDS and MODS (P<0.01) were significantly higher, and the prognosis was poor (P<0.05). CONCLUSION: SIRS is an important factor that has strong influence on incidence of ALI, and the ALI is an important determinant of prognosis and severity of patients with acute pancreatitis. PMID- 14764228 TI - [Effect of peritoneal lavage with povine-iodine on prevention of postoperative sepsis after exposure of peritoneal cavity to sea water in rat]. AB - OBJECTIVE: To observe the effects of peritoneal lavage with povine-iodine on prevention of sepsis after exposure of peritoneal cavity to sea water in rat. METHODS: Eighty-four SD rats were randomly divided into two groups, and the peritoneal cavity was exposed to sea water. Rats in group A were not treated (group A, n=42), and the peritoneal cavity was lavage with povine-iodine in group B (n=42). Plasma levels of endotoxin and tumor necrosis factor (TNF) were measured preimmersion, and 0, 12, 24 hours after seawater immersion (n=6), and positive incidence of blood bacterial culture was performed (n=18 in each group) in groups A and B. RESULTS: 1. Plasma levels of endotoxin and TNF in group A and B were increased significantly after exposure of peritoneal cavity to sea water (compared with baseline values, all P<0.05). Plasma levels of endotoxin and TNF in group B became lower than those in group A from 12 hours after seawater immersion (P<0.05 or P<0.01). 2. Positive incidence of bacterial culture in group B was 16.7 % (3/18) and it was lower than that in group A (77.8 % (14/18), P<0.01). CONCLUSION: Povine-iodine lavage in the peritoneal cavity can reduce levels of plasma endotoxin and TNF, and lower positive incidence of bacterial culture in rats after exposure of peritoneal cavity to sea water, thereby preventing the development of postoperative sepremia. PMID- 14764230 TI - [Effect of continuous renal replacement treatment on stress reaction in patients with severe trauma]. AB - OBJECTIVE: To investigate the effect of continuous renal replacement treatment (CRRT) stresses reaction in patients with severe trauma and its clinical significance. METHODS: Twenty-nine patients with severe trauma were randomly divided into two groups: treatment group (15 cases, treated with continuous veno venous hemofiltration (CVVH) within 12 hours after trauma) and control group (14 cases, not treated with CVVH). The levels of cortical, interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) in plasma were determined at 12 hours, 24 hours, 48 hours, 72 hours after trauma, and the temperature, heart beat, respiratory rate, blood pressure were observed. RESULTS: The levels of cortical, IL-6 in both groups increased to some degree in 12 hours. In control group, the level of cortical increased gradually, peaking at 48 hours, and it was still higher than normal at 72 hours. The level of cortical decreased in the treatment group gradually after 24 hours, approaching normal level at 72 hours. When compared with the control group, the levels of cortical in plasma were lower obviously in the treatment group at 24 hours, 48 hours, 72 hours (all P<0.001). The level of IL-6 decreased obviously after 12 hours and was lower evidently at 24 hours,48 hours, 72 hours (all P<0.05). The clinic symptoms were improved obviously. The level of TNF-alpha was increased at each time point in both groups, but there was no significant difference between two groups. CONCLUSION: CRRT can eliminate effectively the stress hormone, and reduce stress reaction obviously. The change in IL-6 is more marked than that of cortical. PMID- 14764231 TI - [Changes in myocardium enzyme in rabbits with acute renal failure]. PMID- 14764241 TI - Analysis of the essential oils of the leaves, stems, rhizomes and roots of the medicinal plant Alpinia galanga from southern India. AB - The essential oils of the leaves, stems, rhizomes and roots of the medicinal plant Alpinia galanga from southern India were investigated by GC-FID, GC-MS and olfactometry. In all four samples, mono- and sesquiterpenes as well as (E)-methyl cinnamate could be identified. They are responsible for the characteristic odor as well as for the reported use in (folk) medicine and in food products of A. galanga. The essential oil of A. galanga leaves is rich in 1,8-cineole (28.3%), camphor (15.6%), beta-pinene (5.0%), (E)-methyl cinnamate (4.6%), bornyl acetate (4.3%) and guaiol (3.5%). The stem essential oil contains 1,8-cineole (31.1%), camphor (11.0%), (E)-methyl cinnamate (7.4%), guaiol (4.9%), bornyl acetate (3.6%), beta-pinene (3.3%) and alpha-terpineol (3.3%). 1,8-cineole (28.4%), alpha fenchyl acetate (18.4%), camphor (7.7%), (E)-methyl cinnamate (4.2%) and guaiol (3.3%) are the main constituents of the rhizome essential oil. The root essential oil contains alpha-fenchyl acetate (40.9%), 1,8-cineole (9.4%), borneol (6.3%), bornyl acetate (5.4%) and elemol (3.1%). In addition, biological and aroma effects of the main and minor compounds of the four essential oils of Alpinia galanga are discussed in terms of their possible use in medicine, cosmetics and foods. PMID- 14764242 TI - Electrothermal atomic absorption spectrometric-determination of cobalt in human serum and urine. AB - Problems and possibilities of the determination of Co in serum and urine samples by electrothermal atomic absorption spectrometry (ETAAS) are described. Optimal instrumental parameters as well as a suitable atomizer, calibration procedure and hydrogen peroxide as modifier are proposed for direct ETAAS measurement of Co in serum and urine. The detection limit achieved was 0.1 microg L(-1) for both matrices and relative standard deviations varied in the range 5-20% depending on the Co concentration in the sample. Validity of the method was verified by the analyses of standard reference materials. For serum samples with a Co content lower than the detection limit, a separation and preconcentration procedure based on liquid/liquid extraction is suggested prior to determination of Co in the organic phase by ETAAS. This procedure permits determination of 0.02 microg L(-1) Co in serum samples with a relative standard deviation of 10-18%. PMID- 14764243 TI - Hepatoprotection by 3-bromo-6-(4-chlorophenyl)-4-methylthio-2H-pyran-2-one against experimentally induced liver injury in rats. AB - Hepatoprotective activity of 3-bromo-6-(4-chlorophenyl)-4-methylthio-2H-pyran-2 one, an isostere of dimethyl ricinine, was evaluated in adult male albino rats intoxicated with carbon tetrachloride, paracetamol or thioacetamide. The test compound showed significant hepatoprotection at 6.0 mg kg(-1) body mass daily dose, given to the rats for seven consecutive days. The carbon tetrachloride, paracetamol and thioacetamide were given, respectively, on days 3, 5, and 7, on day 6 and on day 6 post treatment with the test compound. The protective effect was evident in a battery of serum and liver biochemical parameters related to hepatotoxicity. PMID- 14764244 TI - Development and in vitro evaluations of gelatin A microspheres of ketorolac tromethamine for intranasal administration. AB - Gelatin A microspheres (MS) of ketorolac tromethamine (KT) for intranasal systemic delivery were developed with the aim to avoid gastro-intestinal complications, to improve patient compliance, to use as an alternative therapy to conventional dosage forms, to achieve controlled blood level profiles, and to obtain improved therapeutic efficacy in the treatment of postoperative pain and migraine. Gelatin A microspheres were prepared using the emulsification crosslinking technique. The drug was dispersed in polymer gelatin and formulated into a w/o emulsion with liquid paraffin, using glutaraldehyde as a crosslinking agent. The formulation variables were drug loading and the concentrations of polymer (gelatin), co-polymer (chitosan) and the crosslinking agent. All the prepared microspheres were evaluated for physical characteristics, such as particle size, incorporation efficiency, swelling ability, in vitro bioadhesion on rabbit small intestine and in vitro drug release characteristics in pH 6.6 phosphate buffer. All the microspheres showed good bioadhesive properties. Gelatin A and chitosan concentrations, percentage of the crosslinking agent and also the drug loading affected significantly the rate and extent of drug release. The data indicated that the KT release followed Higuchi's matrix model. PMID- 14764245 TI - Toxicovigilance: new biochemical tool used in sulfonylurea herbicides toxicology studies. AB - In vitro toxic effects of sulfonylurea herbicides (thifensulfuron-methyl and metsulfuron-methyl) were evaluated according to a new protocol. Physiological conditions were reproduced in order to boost toxicovigilance. Sulfonylureas and their hydrolysis products were added to biological substrates such as urea, alanine, aspartic acid, alpha-ketoglutarate, oxaloacetate, pyruvate and then incubated with some specific enzymes. Addition of these sulfonylureas and their degradation products did not significantly change the enzymatic activity of the urease, aspartate-aminotransferase, glutamate dehydrogenase, malate dehydrogenase and lactate dehydrogenase. However, the acid hydrolysis products inhibited up to 95% of the activity of the alanine-aminotransferase at low concentrations (0.27 micromol L(-1)). Inhibition did not affect the mitochondrial aspartate aminotransferase. PMID- 14764246 TI - Matrix type transdermal drug delivery systems of metoprolol tartrate: in vitro characterization. AB - The monolithic matrix type transdermal drug delivery system of metoprolol tartrate were prepared by the film casting on a mercury substrate and characterised in vitro by drug release studies, skin permeation studies and drug excipients interaction analysis. Four formulations were developed, which differed in the ratio of matrix-forming polymers. Formulations MT-1, MT-2, MT-3 and MT-4 were composed of Eudragit RL-100 and polyvinyl pyrrolidone K-30 with the following ratios: 2:8, 4:6, 6:4 and 8:2, respectively. All the four formulations carried 10% (m/m) of metoprolol tartrate, 5% (m/m) of PEG-400 and 5% (m/m) of dimethyl sulfoxide in isopropyl alcohol: dichloromethane (40:60). Cumulative amounts of the drug released in 48 hours from the four formulations were 61.5, 75.4, 84.3 and 94.5%, respectively. The corresponding values for cumulative amounts of the permeated drug for the said formulations were 53.5, 62.5, 69.8 and 78.2%. On the basis of in vitro drug release and skin permeation performance, formulation MT-4 was found to be better than the other three formulations and it was selected as the optimized formulation. PMID- 14764247 TI - Design and synthesis of some new derivatives of 3H-quinazolin-4-one with promising anticonvulsant activity. AB - A new series of 3-substituted (methyl, ethyl or phenyl)-3H-quinazolin-4-one derivatives (4a-l) were synthesized through condensation reaction of their potassium salts (3a-l) with methyl, ethyl and phenylisocyanate. The newly synthesized derivatives (4a-l) were evaluated for anticonvulsant activity. It was found that these compounds showed the highest anticonvulsant activity at low doses (50-100 mg kg(-1)), whereas at doses over 100 mg kg(-1) they showed a stimulant effect on the central nervous system that even potentiated the effect of the convulsive agent, pentylenetetrazole, in mice. A series of halogenated derivatives, 3-methyl, 3-ethyl and 3-phenyl-6-mono and 6,8-disubstituted-3H quinazolin-4-one derivatives (4m-z) was also synthesized and evaluated for anticonvulsant activity. Reduced anticonvulsant activity was recorded. Phenobarbitone sodium was used as a reference drug. PMID- 14764248 TI - Quantitative analysis of the polyphenols of the aerial parts of rock samphire- Crithmum maritimum L. AB - Quantitative spectrometric analyses of the contents of flavonoids, tannins and total polyphenols in the aerial parts of rock samphire--Crithmum maritimum L., collected from three separated geographic locations along the Adriatic coast of Croatia in different growth stages, are reported. The results point to variability of the analyte content as a function of the growing site and growth stage. The content of flavonoids was found to be 0.08-0.42%. The highest content of flavonoids was found in the samples collected before flowering. The content of tannins ranged from 0.10 to 2.65%, while the content of total polyphenols varied from 4.72 to 9.48%. The highest contents of tannins and total polyphenols were found in the samples collected before flowering and at the beginning of flowering. PMID- 14764249 TI - External flavonoid aglycones from Veronica chamaedrys L. (Scrophulariaceae). AB - External flavone aglycones in the overground parts of Veronica chamaedrys L. (Scrophulariaceae) have been analysed by isocratic reversed-phase high performance liquid chromatography. Luteolin, apigenin, luteolin-3'-methyl ether and scuterallein-6,4'-dimethyl ether have been identified. The latter compound is a new flavonoid identified in Veronica genus. PMID- 14764250 TI - Standardization of biomedical journals in Pakistan. PMID- 14764251 TI - Evolution of biomedical research publications and indexation in Pakistan. PMID- 14764252 TI - Outcome of cardiogenic shock complicating acute myocardial infarction. AB - OBJECTIVE: To analyze the characteristics and in-hospital outcome of patients with cardiogenic shock (CS) complicating acute myocardial infarction (AMI) and to evaluate the influence of urgent coronary revascularization on in-hospital mortality. DESIGN: Descriptive study. PLACE AND DURATION OF STUDY: The Aga Khan University Hospital, Karachi. January 2001 to December 2001. MATERIALS AND METHODS: All consecutive patients with AMI and CS, admitted at The Aga Khan University Hospital, Karachi Pakistan, during the year 2001 were reviewed. A pre designed questionnaire was used for data collection. Analysis was done using the SPSS statistical package. RESULTS: Out of 615 patients with AMI, 53 (8.6%) had CS. Mean age was 60.9 +10.7 years. 62.3% were men, 52.8% were hypertensive and 43.4% were diabetic. Most infarcts were anterior in location (56.6%). Thrombolytic therapy (Streptokinase) was administered to 43.5% of patients with ST segment elevation myocardial infarction. 64.2% required ventilatory support while swan ganz was used in 37.7%. Intra-aortic balloon pump was inserted in 39.6%. Ventricular tachycardia was the most common complication (39.6%). Overall in-hospital mortality was 54.7%. CS associated with mechanical complications had 80% in-hospital mortality. In patients without mechanical complications (n=48), in-hospital mortality was significantly lower in the revascularization group (31.6% vs. 65.5%, p-value = 0.021). However, there were significant differences in the baseline characteristics in the two groups because of the selection bias. CONCLUSION: CS occurring in patients with AMI has an extremely poor prognosis. Patients selected for revascularization strategy has favorable in-hospital outcome. PMID- 14764253 TI - Magnetic resonance imaging in the diagnosis and local staging of primary bone tumors. AB - OBJECTIVE: To evaluate relationship of magnetic resonance (MR) imaging in the diagnosis and local staging of primary bone tumors with histopathological and surgical outcome. DESIGN: Cross-sectional comparative study. PLACE AND DURATION OF STUDY: Radiology Department, The Aga Khan University Hospital, Karachi from December 1999 to March 2002. SUBJECTS AND METHODS: Thirty patients suspected of having primary bone tumors were selected. MRI scans were evaluated for local staging including intramedullary tumor length, cortical, soft tissue, neuro vascular, joint and epiphyseal involvement. MR staging was compared with surgical and pathological staging and MR diagnosis with histopathological diagnosis considering benign and malignant tumors and specific diagnosis. RESULTS: The study included 30 patients with age ranging from 5 - 75 years (mean 30.1 years). Twenty-three patients had malignant and 7 benign tumors. On MRI, 25 tumors were characterized as malignant and 2 tumors were false positive for malignancy. CONCLUSION: MR imaging proved to be accurate in local staging of bone tumors and determination of the extent of disease in benign tumors. PMID- 14764254 TI - Dehydration related abdominal pain (DRAP). AB - OBJECTIVE: To describe the frequency of dehydration as a medical cause of acute abdomen. DESIGN: Cross-sectional analytical study. PLACE AND DURATION OF STUDY: The study was conducted at Combined Military Hospital, Malir Cantonment Karachi between March 1, 2000 to March 1, 2002. SUBJECTS AND METHODS: All the patients reporting with abdominal pain to the surgical outpatient department or the emergency department, were reviewed in the study. The clinical findings in all these cases were studied along with the mode of their management and outcome. RESULTS: Of all the patients presenting with abdominal pain, 303% (n=68) were suffering from dehydration related abdominal pain. They were predominantly males in a ratio of 8.7:1, mostly in the 2nd and 3rd decades of their lives. All these cases were suffering from acute or chronic dehydration were provisionally diagnosed by general practitioners as 'acute abdomen' and referred for surgical consultation. Associated symptoms included vomiting in 42.6% backache in 91.2%, headache in 95.6%, and pain in lower limbs in 97.1% of the cases. 83.8% required indoor management with intravenous fluids. All the patients became asymptomatic with rehydration therapy. CONCLUSION: Dehydration is a possible cause of severe abdominal pain. There is a need to educate the general public about the benefits of adequate fluid intake. PMID- 14764255 TI - Correlation of HBV DNA PCR and HBeAg in hepatitis B carriers. AB - OBJECTIVE: To correlate hepatitis B HBV DNA polymerase chain reaction (PCR) results with HBeAg and serum alanine transferase (ALT) in carriers. DESIGN: Non interventional descriptive study. PLACE AND DURATION OF STUDY: The study was carried out at Virology Department, Armed Forces Institute of Pathology (AFIP), Rawalpindi, from January 2001 to September 2002. MATERIALS AND METHODS: Fifty hepatitis B carriers, with known HBsAg positive serostatus, raised serum ALT and detectable HBV DNA, were selected out of the patients reporting at AFIP for their blood test for HBV DNA. HBV DNA testing in these cases was carried out using PCR kit of Accugen-USA. After confirmation of their carrier status and raised serum ALT levels, the sera were tested for HBeAg and results of HBeAg testing were correlated with those of HBV DNA testing. RESULTS: Out of the total 50 HBV DNA PCR positive hepatitis B carriers, 48 samples were positive for HBeAg. All the 50 HBV DNA positive cases had raised serum ALT levels. CONCLUSION: In case of non availability of facility for HBV PCR, detectable HBeAg should be taken as a surrogate marker for HBV DNA in hepatitis B carriers with raised serum ALT. PMID- 14764256 TI - Efficacy and tolerability of tegaserod in constipation dominant irritable bowel syndrome. AB - OBJECTIVE: To determine the efficacy and tolerability of tegaserod in the treatment of symptoms of irritable bowel syndrome (IBS) IBS-C patients. DESIGN: An open label (quasi interventional) study. PLACE AND DURATION OF STUDY: Patients were enrolled between October 2000 and August 2001 at 4 centres (AKUH, Karachi; Mayo Hospital, Lahore; PIMS, Islamabad; Hayatabad Teaching Complex, Peshawar). PATIENTS AND METHODS: Tegaserod was administered in a dose of 6 mg (twice-a-day) orally for a period of 6 weeks. Symptoms were assessed before and during treatment using a questionnaire. RESULTS: The mean age of patients was 37.5 years and 81(69.2%) were males. The study enrolled 117 patients and 101 patients completed the study. Number of bowel movements, symptoms of straining at defecation, stool consistency, bloating, urgency and abdominal pain improved significantly following treatment (p<0.05). Analysis of data in both genders separately showed statistically significant improvement in symptoms of urgency, straining at defecation, abdominal pain and number of bowel movements following treatment. Side effects of diarrhoea and vertigo (6 and 1 patients respectively) necessitating discontinuation of treatment were infrequent. CONCLUSION: Tegaserod given in a dose of 6 mg b.d. is effective and well tolerated in IBS-C patients. It is equally effective in males and females in relieving the symptoms of abdominal pain, bloating, straining at defecation as well as increased in the mean number of bowel movements per week. PMID- 14764257 TI - Causes and management of postoperative enterocutaneous fistulas. AB - OBJECTIVE: To identify the causes of postoperative enterocutaneous fistulas and to evaluate the results of conservative and operative treatment including the effectiveness of octreotide in the management of these fistulas. DESIGN: A descriptive study. PLACE AND DURATION OF STUDY: Department of Surgery, Liaquat University of Medical and Health Sciences, Jamshoro / Hyderabad between January 1997 and December 2001. SUBJECTS AND METHODS: Forty patients with postoperative fistula were studied. Demographic variables, causes and management outcome was observed and recorded. RESULTS: There were 25 males and 15 females with 50% of the patients being in age group of 21-30 years. Emergency surgery for typhoid perforation(45%) and intestinal tuberculosis (30%) were the commonest causes. Ileum and jejunum were the commonest sites of fistulation found in 85% cases. Twenty-one patients were started on conservative treatment with spontaneous closure occurring in 15 (71.4%) patients. Nineteen patients were operated within three days of admission due to generalized peritonitis (73.7%) and local intra abdominal collections (26.3%). Wound infection was the commonest complication in the operative group. The mortality rate in this series was 7.5%. All the deaths occurred following surgery. CONCLUSION: Postoperative enterocutaneous fistula has a high morbidity and a significant mortality. Sepsis in the peritoneal cavity is the major cause of mortality. Conservative treatment has a good outcome for these fistulas. The use of octreotide is highly recommended as it definitely converts high output fistulas to low output fistulas. PMID- 14764258 TI - Management of oromandibular cancers. AB - OBJECTIVE: To emphasize the role and importance of multidisciplinary approach in the management of oral cavity cancers involving the mandible. DESIGN: Descriptive study. PLACE AND DURATION OF STUDY: The Departments of Plastic and Reconstructive Surgery, ENT / Head and Neck Surgery and Radiation Oncology, Combined Military Hospital, Rawalpindi. Duration spans over a period of six years. SUBJECTS AND METHODS: A total of 63 patients who had biopsy-proven oromandibular tumors, after thorough assessment / staging in Joint Head and Neck Oncology Clinic, underwent resection and reconstruction for malignant oral cavity tumors involving the mandible were included in the study. All the resected tumor specimen were sent for histopathology. All the post-resection defects were properly classified and reconstructed by the plastic surgery team. Postoperatively, all the patients underwent adjuvant full dose radiotherapy at the Department of Radiation Oncology. Complications were recorded and managed accordingly. At one year follow up all the available patients were assessed for functional and aesthetic restoration and recurrences. RESULTS: Out of 63 patients there were 40 males and 23 females (ratio 1.7 : 1) with an average age of 50 years. Tumor-free resection margins could be achieved in 56 patients. In 88% cases tumor was a Squamous cell carcinoma. Radical neck dissections were carried on in 27 patients. Radial forearm free flap was used in 27 patients, pectoralis major myocutaneous flap in 19, free fibula osteocutaneous flap in 10, rectus-abdominis myocutaneous free flap with Implant was used in 3 patients to reconstruct the post-resection defects. There was only one total flap loss and 3 partial flap losses. Implant exposure was encountered in 4 instances with 3 major and 5 minor fistulae. At one year follow-up, 56 patients were available. Thirty-seven patients had intelligible speech, 15 patients were taking normal diet and in 33 patients there was a satisfactory mandibular contour restoration. Seven patients had recurrences, 2 were not traceable and 5 patients had died by that time. CONCLUSION: A multidisciplinary collaboration is the key to effectively manage this group of extremely debilitating malignancies. PMID- 14764259 TI - Demographic and Clinicopathologic features of membranous glomerulonephritis. AB - OBJECTIVE: To determine the frequency of membranous glomerulonephritis(MGN) in the renal biopsy specimens. To observe the presenting features and the demographics to determine association, if any, between MGN and hepatitis B in these patients. DESIGN: Descriptive study. PLACE AND DURATION OF STUDY: The section of histopathology, The Aga Khan University Hospital, Karachi over a period of three years (May 1999-April 2002). SUBJECTS AND METHODS: All consecutive percutaneous renal biopsy specimens received during the study period were included. A total of 1590 renal biopsy specimens were evaluated. All cases where a definitive diagnosis of MGN (stage II to IV) was made on light and/or immunofluorescence studies were included in the results. Twenty-eight cases were excluded owing to inadequate material. Cases with a presumptive diagnosis and those in stage I of MGN were also excluded. Laboratory investigations carried out include urine examination, blood urea nitrogen. (BUN), serum creatinine, 24 hours urinary protein, Hepatitis B sAg, serological tests for hepatitis C virus, ANA, dsDNA, etc. Clinical features and results of laboratory analysis were recorded along with light microscopic features and statistical analysis was carried out. RESULTS: Out of 1562 renal biopsy specimens, 835(53.4%) cases showed primary glomerular pathology. 741(47.4% of the total) cases presented with nephrotic syndrome. Out of these in 176 cases (23.75%), findings were those of membranous glomerulonephritis followed by amyloidosis and focal segmental glomerulosclerosis. Among the cases with MGN, 23(13%) cases were those of secondary MGN whereas 87% were grouped under primary MGN. The male to female ratio was 2.2:1. Mean age at presentation was 29 years, median 29 years and a mode of 30 years. Nineteen cases (1.07%) were seen in children of 14 years old or less. CONCLUSION: The frequency of MGN was much higher in our study subjects as compared to most other regional studies. No significant association with hepatitis B was seen with only 4 out of 89 cases showing positivity for HBsAg. The pattern of disease corresponds more to that seen in the west. PMID- 14764260 TI - Malignant hyperthermia. AB - Malignant hyperthermia refers to covert myopathies, which do not affect the individual during daily life activities, but may lead to life-threatening tachycardia, rigor, labile blood pressure and most importantly high-grade temperature when exposed to general anaesthesia. This conditions is mimicked by thyroid storm, neuroleptic malignant syndrome, phaeochromocytoma and sepsis. We present a presumptive case of malignant hyperthermia. PMID- 14764261 TI - Cardiac trauma with gunshot injuries. AB - Penetrating cardiac injuries, secondary to gunfire, constitute the most lethal forms of cardiothoracic trauma with their potential fatality. We report our experience of managing two such cases who presented with haemorrhagic shock and cardiac tamponade, in a collapsed state. Prompt resuscitation and early surgical intervention (midline sternotomy and cardiorrhaphy) was successfully performed with a favourable outcome. PMID- 14764262 TI - Morgagni's hernia. AB - Congenital diaphragmatic hernias, including Morgagni s hernia, usually present in early childhood and are treated by surgical repair. This case report is about an unusual Morgagni s hernia, presenting with dyspepsia and chest pain, at the age of 45 years. For many years the diagnosis remained a dilemma because patient s chest x-ray was not done and she was treated for "angina " and "dyspepsia". Diagnosis was obvious once a chest x-ray was done, however, barium studies were performed for further confirmation. PMID- 14764263 TI - Extradural Granulocytic sarcoma causing acute paraparesis. AB - A case of 9 years old female presenting with rapidly progressive paraparesis during remission phase of acute myeloblastic leukemia is reported. Radiological imaging revealed an extradural mass in the upper dorsal spine producing significant cord compression. The patient showed a dramatic neurological recovery after spinal cord decompression and subsequently treated with appropriate chemotherapy and local radiotherapy. PMID- 14764264 TI - Cardiac arrhythmia as presentation of snakebite. AB - Snakebite cases may have myriad presentations. we are describing a previously healthy young man presenting within half an hour of snakebite who experienced abrupt fatal cardiac rhythm changes ranging from bradycardia to ventricular tachycardia/fibrillation over a short span of time. PMID- 14764265 TI - Ectopic ureters misdiagnosed as ureterocele. AB - Ectopic ureters may be difficult to diagnose unless there is high index of suspicion. Examination for continuous dribbling may be difficult in infants who are still using diapers. We are reporting a case of ectopic ureter which was erroneously diagnosed as ureterocele on a cystourethrogram. Further investigations for accurate diagnosis and emerging abnormalities during the course of management are presented in this case report. Review of literature to highlight the various forms of investigations and management are also presented. PMID- 14764266 TI - Pseudotumor of urinary bladder. AB - The case of an old male is presented who had postoperative spindle cell nodule of urinary bladder. This benign condition mimics a sarcoma. However, the treatment modalities and the outcome are entirely different. Inflammatory pseudotumor/PSCN and leiomyosarcoma of the bladder overlap in clinical and immunohistochemical findings but are histologically as well as clinically distinct, correlating with different natural histories. Inflammatory Pseudotumor/PSCN are benign, not prone to metastasize, and is outside the spectrum of low-grade inflammatory sarcoma of urinary bladder. PMID- 14764267 TI - Urethral diverticulum in female. AB - A case of 50 years old female with a urethral diverticulum is presented. The diagnosis was delayed for many years, it was successfully treated by performing transvaginal diverticulectomy. PMID- 14764268 TI - Hepatocellular carcinoma: available diagnostic tools and their limitations. PMID- 14764269 TI - Spectrum of injuries at the emergency department of a tertiary care hospital. PMID- 14764270 TI - Prevalence and incidence of disease in a hospital-based study. PMID- 14764271 TI - Clinical challenges: the intersection of diabetes, chronic kidney disease, and cardiovascular disease. PMID- 14764272 TI - Acarbose and the risk of cardiovascular disease and hypertension. PMID- 14764273 TI - Beta blockers and their effect on diabetic patients with heart failure. PMID- 14764274 TI - Stent restenosis and the use of drug-eluting stents in patients with diabetes mellitus. AB - Stents have become the technique of choice for percutaneous revascularization, but in-stent restenosis has remained a clinical challenge. This brief article summarizes the incidence, patterns, and proposed mechanisms of restenosis and outlines its contemporary management with specific focus on the diabetic patient. It includes a synopsis of the strategy of drug-eluting stents, which is the most recent and major advance in percutaneous coronary intervention. PMID- 14764275 TI - The role of electron beam computed tomography for measuring coronary artery atherosclerosis. AB - It is generally agreed that the coronary artery calcium score provides significant information for the prediction of future cardiac events. However, additional information is needed to determine whether or not this information is significantly greater than that available from the analysis of established risk factors. Until this is resolved, it is unlikely that electron beam computed tomography can be justified for routine evaluation of patients. However, measurement of coronary artery calcium may be valuable in selected clinical situations, and has potential as a research tool for studying the pathogenesis or treatment of stable coronary artery disease. PMID- 14764276 TI - Are there specific components of the insulin resistance syndrome that predict the increased atherosclerosis seen in type 2 diabetes mellitus? AB - Cardiovascular disease is often present in patients with newly diagnosed type 2 diabetes. It is hypothesized that both type 2 diabetes and cardiovascular disease arise from an underlying insulin resistance syndrome, including impaired glucose tolerance, hypertension, dyslipidemia, and abdominal obesity. We reviewed the literature using various epidemiologic techniques to evaluate the relation between the insulin resistance syndrome components and incident cardiovascular disease. We found that although insulin resistance is likely important in the development of cardiovascular disease seen in diabetes, lipid and blood pressure abnormalities associated with the syndrome appear to be equally important. Future studies are needed to determine the most effective treatment strategies for preventing cardiovascular disease associated with the insulin resistance syndrome and type 2 diabetes. PMID- 14764278 TI - BNP levels to screen for left ventricular dysfunction. PMID- 14764277 TI - The role of advanced glycation end products in the development of atherosclerosis. AB - Cardiovascular disease (CVD) is the leading cause of mortality worldwide. Atherosclerosis constitutes the main pattern of cardiovascular disease and leads to thickening of the intima with plaque formation and eventual occlusion of the arterial lumen. A large amount of evidence links advanced glycation end products (AGEs) with the development or progression of atherosclerosis, regardless of the diabetic status. AGEs are a heterogenous group of compounds formed by the nonenzymatic reaction of reducing sugars with proteins, lipids, and nucleic acids. Although AGEs are formed endogenously in the body, diet has recently been recognized as an important exogenous source. An increased understanding of the mechanisms of formation and interaction of AGEs has allowed the development of several potential anti-AGE approaches. PMID- 14764279 TI - Hyperglycemia with antipsychotic treatment. AB - Diabetes and mental disorders are common chronic illnesses in the United States. Recently, the introduction of a new class of atypical antipsychotic medications has been a major treatment advance for patients with mental disorders. Because of increased use of atypical antipsychotic medications, new and unanticipated side effects have often appeared. Treatment-emergent diabetes has been described for conventional and atypical antipsychotics. People with schizophrenia may be at increased risk for type 2 diabetes because of the side effects of antipsychotic medication, underlying predisposition, and less healthy lifestyles. PMID- 14764280 TI - Periodontal disease and diabetes mellitus. AB - Infections of the tissue surrounding the teeth (periodontitis) are usually caused by anaerobic gram-negative microorganisms. This infection causes destruction of the supporting alveolar bone and can lead to tooth loss. Removal of these microorganisms can slow or arrest the progression of periodontitis. Diabetes patients are at greater risk of developing periodontitis, may not respond as well to periodontal therapy as nondiabetic patients, and may require more aggressive treatment to manage periodontitis. Microorganisms that cause periodontitis and the host response to these may increase insulin resistance in diabetic patients. Treatment of periodontitis could improve glycemic control. A model is presented in which periodontal pathogens may cause increases in proinflammatory cytokines that mediate increases in insulin resistance, resulting in an increase in blood glucose. Following periodontal therapy, this process may be reversed. PMID- 14764281 TI - The metabolic syndrome in children and adolescents. AB - The metabolic syndrome was recently defined by the Adult Treatment Panel III. Despite a lack of uniform definition of the syndrome in pediatrics, recent studies have shown that the syndrome develops during childhood and is highly prevalent among overweight children and adolescents. The hypothesized central role of insulin resistance and obesity as a common underlying feature of the metabolic syndrome also appears to be already manifested in childhood. In view of the current obesity epidemic in children and adolescents, there is a vital need to provide adequate guidelines for the definition of the metabolic syndrome in pediatrics and for the development of screening and treatment strategies. This article focuses on the above issues, as well as on the impact of the syndrome on two major disease outcomes, type 2 diabetes and cardiovascular disease. PMID- 14764282 TI - Cardiovascular risk associated with the metabolic syndrome. AB - The term metabolic syndrome refers to a clustering of cardiovascular risk factors, most of which also share insulin resistance as an additional feature. Scientific effort has concentrated on understanding why these diverse cardiovascular risks co-occur in individuals and in determining the presumed common environmental or genetic factors that might underpin this. Clinically important developments include publication of standard definitions of the metabolic syndrome and recommendations for the use of type 2 diabetes and the presence of the metabolic syndrome as critical "risk stratifiers" in cardiovascular disease prevention. The remarkable recent secular increases in the prevalence of type 2 diabetes and obesity in many populations mean that the importance of the metabolic syndrome as a determinant of cardiovascular disease is likely to increase until these trends can be reversed. PMID- 14764285 TI - Candesartan reduces cardiovascular death in CHF patients on ACE inhibitor. PMID- 14764283 TI - Treatment of PCOS with metformin and other insulin-sensitizing agents. AB - A significant proportion of women with polycystic ovary syndrome (PCOS) suffer from insulin resistance and compensatory hyperinsulinemia. Growing evidence indicates that elevated serum insulin induces hyperandrogenism, which in turn leads to anovulation and infertility. Hyperinsulinemia also contributes to the increased risk for cardiovascular disorders and type 2 diabetes mellitus. These concepts provide a rationale for therapies focused on treatments of insulin resistance. Metformin is the most extensively studied insulin-sensitizing agent for the treatment of women with PCOS. Use of metformin leads to a decrease in serum insulin and androgen levels, as well as an improvement in ovulatory function. Other insulin-sensitizing agents studied in women with PCOS include troglitazone, rosiglitazone, pioglitazone, and D-chiro-inositol. PMID- 14764286 TI - Vaccinations containing thimerosal do not increase rates of autism. PMID- 14764287 TI - Antidepressant treatment reduces poststroke mortality. PMID- 14764288 TI - Azithromycin (3 days) better than amoxicillin-clavulanate (10 days) for sinusitis? PMID- 14764289 TI - Four-year prostate cancer screening interval is effective. PMID- 14764290 TI - Treatment of high LDL saves lives of those with diabetes or cardiovascular disease. PMID- 14764291 TI - Conjunctivitis: Diagnostic usefulness of signs and symptoms unknown. PMID- 14764292 TI - Relative risks and odds ratios: what's the difference? PMID- 14764293 TI - Simplifying the language of evidence to improve patient care: Strength of recommendation taxonomy (SORT): a patient-centered approach to grading evidence in medical literature. AB - Several taxonomies exist for rating individual studies and the strength of recommendations, making the analysis of evidence confusing for practitioners. A new grading scale-the Strength of Recommendation Taxonomy (SORT)-will be used by several family medicine and primary care journals (required or optional), allowing readers to learn 1 consistently applied taxonomy of evidence. SORT is built around the information mastery framework, which emphasizes the use of patient-oriented outcomes that measure changes in morbidity or mortality. Levels of evidence from 1 to 3 for individual studies also are defined. An A-level recommendation is based on consistent and good-quality patient-oriented evidence; a B-level recommendation is based on inconsistent or limited-quality patient oriented evidence; and a C-level recommendation is based on consensus, usual practice, opinion, disease-oriented evidence, or case series for studies of diagnosis, treatment, prevention, or screening. PMID- 14764294 TI - Conjunctivitis and cervicitis. PMID- 14764295 TI - An ounce of prevention: the evidence supporting periconception health care. PMID- 14764296 TI - How should we diagnose and treat osteoarthritis of the knee? PMID- 14764297 TI - Nasal irrigation as adjunctive care for acute sinusitis. PMID- 14764298 TI - Clinical inquiries. Is combining ACE inhibitors and ARBs helpful or harmful? PMID- 14764299 TI - Clinical inquiries. When should we treat isolated high triglycerides? PMID- 14764300 TI - Clinical inquiries. Does treating diastolic BP to less than 90 mm Hg lower cardiovascular risk? PMID- 14764301 TI - Clinical inquiries. Which healthy adults should take aspirin? PMID- 14764302 TI - Clinical inquiries. Should we screen adults for asymptomatic microhematuria? PMID- 14764303 TI - Clinical inquiries. What is the best treatment for Osgood-Schlatter disease? PMID- 14764304 TI - Diagnosis and management of migraine in family practice. PMID- 14764305 TI - Evaluation of non-pharmacological intervention on antisocial behavior in patients suffering from Alzheimer's disease in a day care center. AB - Psychosocial treatment was studied on improving socially non integrated behavior and in containing cognitive-behavioral degeneration in patients suffering from Alzheimer's disease. The hypothesis was that patients who do not maintain social relationships decline in their psycho-social and psycho-cognitive behavior more rapidly and that intervention helps to attain the above mentioned objectives (18 patients in day care center). All subjects had to perform the Cohen-Mansfield Agitation Inventory and the Cornell Depression Scale at the beginning of the study and were tested again after 1.5 years. The results of the group of Alzheimer patients showed a significant difference in the CMAI Scale (P<0.01) and sub-scales and in the Cornell Scale (0.014). In conclusion, the usefulness of the center is fundamental in encouraging the maintenance of significant relationships and helping patients adapt to their disease. PMID- 14764306 TI - Age-related changes in redox status of rat serum. AB - Aging and age-related diseases are known to be associated with increased oxidative stress. To protect against the deleterious effects of oxidative stress, a well-co-ordinated network of enzymatic and nonenzymatic anti-oxidant defense systems is essential. In the present study, we investigated the age-related redox status of serum by analyzing hydrogen peroxide, hydroxyl radical, superoxide scavenging abilities, and other redox markers. Results showed the anti-oxidative capacity to be significantly decreased in serum of aged rats, which was accompanied by a marked increase in peroxide levels. Our analyses also revealed that levels of nitrated proteins, induced by peroxynitrite treatment, were higher in old rats than in young rats. Our results clearly indicated that the serum redox balance shifted toward oxidation during aging. To further confirm this age related redox shift, we quantified the changes in thiol content. The total thiol level was found to be significantly decreased in the aged group. We also noticed an age-related reduction in serum albumin, which may be partially responsible for the decreased serum thiol levels. A similar pattern can be explained by low levels of serum GSH in old rats compared to young rats. The significance of the present study is the data showing increased oxidative stress in serum during aging, attributed to a decrease in major antioxidant components in serum. PMID- 14764307 TI - Difference in average survival between F344/Du and F344/N rats is not due to genetic contamination. AB - The average survival was found to be different between substrains of rats, F344/Du and F344/N. One of the causes of this difference could have been genetic contamination by crossbreeding with other strain(s) after the divergence of substrains. These two strains were examined by PCR-SSLP (simple sequence length polymorphism) procedure using microsatellite (Mit) markers to identify the contamination. All 288 Mit markers distributed throughout the genome exhibited no size polymorphisms between these two strains. This indicated that there was no genetic contamination between these two strains. The phenotypic difference in average survival, thus, is not due to genetic contamination. PMID- 14764308 TI - Is routine screening for urinary tract infection in rehabilitation day-hospital elderly patients necessary? AB - It is not known whether all rehabilitation day-hospital patients need to be assessed for urinary tract infection (UTI). The aims of this study were to identify patients at high risk to develop UTI and to determine whether there was an association between the amount of post-void residual urine (PVR) and UTI. We surveyed 211 consecutive patients admitted to the day-hospital unit of a geriatric rehabilitation center between June 1998 and February 1999. The main diagnoses were stroke, orthopedic surgeries and deconditioning. Urine samples for general analysis and bacteriology were collected from each patient upon admission. PVR was measured using portable ultrasound. Telephone interviews were conducted with 141 patients, 3-11 months after the initial screening. The prevalence of UTI at admission was higher in women (P=0.007), and patients with decreased functional level (P=0.001). The incidence rate correlated with the functional level but not with gender or main diagnosis. The relative risk of dependent patients to develop UTI was 7.5 times higher than in independent patients. Significant amounts of PVR were equal in males and females. The relative risk of individuals with significant amounts of PVR to develop UTI was 1.63 times greater then those with negligible and moderate amounts of PVR. Gender, degree of functional level and main diagnosis had no effect on the relationship between PVR and UTI. We conclude that low functional level and significant amounts of PVR are risk factors for prevalence and incidence of UTI in elderly patients in a rehabilitation day-hospital. There is no need for routine urine analysis in every elderly patient admitted to rehabilitation day hospitals, however, PVR measurements by portable ultrasound are valuable. PMID- 14764309 TI - The curcuma antioxidants: pharmacological effects and prospects for future clinical use. A review. AB - In agreement with the predictions of the oxygen-stress theory of aging and age related degenerative diseases, diet supplementation with a number of phenolic or thiolic antioxidants has been able to increase the life span of laboratory animals, protect against senescent immune decline and preserve the respiratory function of aged mitochondria. In addition to the above, more recent data reviewed here suggest that the polyphenolic compound curcumin and related non toxic antioxidants from the rhizome of the spice plant Curcuma longa have a favorable effect on experimental mouse tumorigenesis as well as on inflammatory processes such as psoriasis and ethanol-caused hepatic injury. Our own research has focused on the effects of diet supplementation with an antioxidant-rich hydroalcoholic extract of the curcuma rhizome on key risk factors of atherogenesis and related cardiovascular disease. Our reviewed data show that, in human healthy subjects, the daily intake of 200 mg of the above extract results in a decrease in total blood lipid peroxides as well as in HDL and LDL-lipid peroxidation. This anti-atherogenic effect was accompanied by a curcuma antioxidant-induced normalization of the plasma levels of fibrinogen and of the apo B/apo A ratio, that may also decrease the cardiovascular risk. The reviewed literature indicates that curcumin and related plant co-antioxidants are powerful anti-inflammatory agents. Further, since they potentiate the anti-atherogenic effect of alpha-tocopherol, more extensive clinical testing of their probable usefulness in cardiovascular risk reduction seems justified. PMID- 14764310 TI - The leptin, a new hormone of adipose tissue: clinical findings and perspectives in geriatrics. AB - Obesity has gained a great importance during the last decades, and this fact stimulated numerous studies regarding the genetic causes of this disease. A recently discovered new molecule, called leptin, raised a wide interest. It is a product of the adipocytes, it exerts inhibitory effects on the center of appetite and increases the energy expenditure of the organism. The present study evaluated blood leptin levels in 57 elderly subjects and searched for eventual correlations between this parameter and the age, the body mass index (BMI), the fat body mass (fat%), the waist (W) and hip (H) circumference, as well as the ratio (R) of these latter two values (WHR). Blood leptin levels do not correlate with age, body height and the WHR, but display significant positive correlations with the body weight, the BMI, the fat%, the W, H and WHR. A deeper knowledge on leptin and the correlations of this hormone with other body parameters might be helpful in a better understanding of several pathogenetic mechanisms related to aging and involved in a deterioration of the quality of life in elderly, like multiple atherosclerotic and metabolic diseases (diabetes, dyslipidemias). PMID- 14764311 TI - Comparing the alcohol-related problems survey (ARPS) to traditional alcohol screening measures in elderly outpatients. AB - Older drinkers may incur alcohol-related risks at low consumption levels, but commonly used screening measures do not address alcohol's effects among persons with declining health and increased medication use. We compared the newly developed Alcohol-Related Problems Survey (ARPS) to three validated alcohol screens: the Cut down, Annoyed, Guilty, Eye-opener (CAGE), Short-Michigan Alcohol Screening Test (SMAST), and Alcohol-Use Identification Test (AUDIT). The ARPS classifies drinking as non-hazardous, hazardous or harmful. Non-hazardous drinking is defined as consumption with no known risks for adverse physical or psychological health events. Hazardous drinking is consumption with such risks. Harmful drinking results in adverse events. The AUDIT screens for hazardous and harmful drinking; the CAGE and SMAST identify abusive (e.g. failure to fulfill social obligations) and dependent (e.g. having withdrawal symptoms) drinkers. In this study of 574 current drinkers 65 years and older who completed the ARPS and AUDIT in primary care clinics, half were randomly assigned to complete the CAGE and half, the SMAST. Drinkers who screened positive on the CAGE, SMAST or AUDIT were correctly classified by the ARPS as hazardous or harmful drinkers 91, 75, and 100% of the time, respectively. The majority of ARPS-identified hazardous or harmful drinkers did not screen positive on the CAGE, SMAST or AUDIT. These drinkers had medical conditions or used medications that placed them at risk for adverse health events, none of which was addressed in these three screens. In this study, the ARPS identified nearly all drinkers detected by the CAGE, SMAST, and AUDIT and detected hazardous and harmful drinkers not identified by these measures. PMID- 14764312 TI - Classifying the probability of urinary incontinence in psychogeriatric nursing home patients. AB - Urinary incontinence (UI) frequently occurs in psychogeriatric nursing home patients. In general the personnel involved in the care for these patients act on incontinence noted. Patients are not monitored or classified according to likelihood or severity of incontinence. This study was conducted to develop and validate a model for the classification of the likelihood of UI in demented nursing home patients. A multi-center cross-sectional study was conducted using data on clinical and functional status of 692 subjects. Subjects were subdivided in a Derivation set of 532 patients and a Validation set of 160 patients. The data were ascertained with questionnaires completed by physicians and nursing staff. All psychogeriatric wards (25) of four Dutch nursing homes were included. Using univariate logistic regression analysis on the derivation set we identified correlates of UI among 22 clinical and functional patient characteristics. Subsequently, we developed a classification model for prevalent UI, including independent patient characteristics by means of multivariable logistic regression. Next, we stratified patients into groups with varying likelihood's of UI based on the model developed. Subsequently, we transformed the model to an easy applicable classification rule for the identification of patient subgroups with high or low likelihood on UI. Finally, the rule was validated on the validation set. The independent multivariate factors associated with urinary incontinence were impaired ADL and mobility, diminished alertness and fecal impaction. After transforming the regression model to an easy classification rule, the scores ranged from 0 to 7. The area under the curve was 0.88 (95% Confidence Interval (CI): 0.85-0.91) in the derivation set. In the validation set a similar area under the curve was obtained (0.90 (95% CI: 0.85-0.95)). Among subjects with none of the associated factors the rule classified 0.5% as incontinent patients. In case all associated factors were present the proportion classified as incontinent increased to 91%. In conclusion, the developed classification rule provides means to stratifying nursing home patients according to their likelihood of being incontinent of urine. PMID- 14764313 TI - Functional disability of older persons in long-term care facilities in Korea. AB - The aim of this study was to examine functional disability of older persons in long-term care facilities and to identify factors associated with their levels of functioning. The first national survey of older residents in nursing and residential care homes in Korea was conducted in 1999. Of the 1105 respondents who completed the interview 73% in residential care homes and 88% in nursing homes reported limitations in functioning. The percentage of persons with three or more limitations in activities of daily living (ADL) was 63% among nursing home residents and 36% among those at residential care homes. Overall, persons of older age, female, widowed, with less family support, and bearing out-of-pocket costs of medical care tended to show higher levels of disability. Presence of arthritis, hypertension, and stroke was found to be significantly associated with increased likelihood of disability. Multivariate analysis revealed variability in the associations by type of facility. Characteristics such as older age, women, and stroke were significantly associated with a higher likelihood of disability for those in both residential and nursing homes. Arthritis, fracture, exercise, and contact with family members, however, were found to be independently associated with disability among those in residential care homes only. Recent government proposals to improve care delivered to long-term care residents need to take into account their differing levels of functional disability and its associated care needs. PMID- 14764314 TI - Doctors' attitudes and end-of-life decisions in the elderly: a comparative study between Sweden, Germany and Russia. AB - The study was performed in order to determine cross-culturally to what extent various attitudes (legal and ethical concerns, hospital costs, level of dementia, patient's age, patient's wishes, family wishes, doctor's religion) influence the treatment decisions of doctors in face of a critically ill incompetent elderly patient. Convenience samples of doctors in Sweden (n=104), Germany (n=191) and Russia (n=232) were surveyed by means of a self-administered questionnaire comprising a case-vignette of an incompetent elderly patient. The importance of various attitudes attributed to the treatment decision varied between countries. The differences were mostly pronounced for the importance of hospital costs (highest in Russia), patient and family wishes (both highest in Sweden) and level of dementia (highest in Sweden). They reflect diverging clinical practice and underlying societal values. They point to the necessity of the development of uniform standards and training of doctors in ethical issues. PMID- 14764315 TI - Alendronate reduces the daily consumption of insulin (DCI) in patients with senile type I diabetes and osteoporosis. AB - The use of Alendronate for the treatment of senile diabetes with osteopenia or osteoporosis is a common practice today, although the reasons for the success of this treatment are not completely understood. We investigated 40 elderly female patients, over 70 years of age, divided in two Groups (A and B) 20 cases of each, with insulin-dependent senile diabetes and fair metabolic balance, with an average disease duration of 30 +/- 4 years. They all had osteoporosis shown by the mean T-score of bone mineral densitometry. The Groups were treated as follows, Group A with 10 mg/day of Alendronate per os, with morning fasting plus a supplementation of calcium and vitamin D3, while the Group B received only calcium and vitamin D3 per os. Bone mineral density (BMD) expressed in mg/cm2, and in terms of T-score and Z-score at the spine (L1-L4) was monitored over time after 12 and 24 months, using dexa technique with a Lunar DPX densitometer. Moreover, the variation of daily consumption of insulin (DCI) of all the study population was calculated 12 and 24 months after the start of treatments. The data of Group A showed an improvement of osteoporosis, as evidenced by the increase of BMD at both times of measurement, accompanied by a significant reduction in the DCI (-21.6% by the 12th month, and -36.2% by the end of the observation period). In the Group B only small, statistically insignificant changes were observed in both the BMD and DCI. The most plausible explanation of reduction of DCI in Group A seems to be that Alendronate has improved the clinical symptoms of osteoporosis (pain, rigidity, and reduction of movements) through its action on the bone mass recovery and slowing down the bone turnover and under these conditions the diabetic patients improved their own physical performance. The better and more extensive movements certainly produced a reduction in the DCI, since a correct and adequate physical activity does contribute to an improved glucose metabolism. PMID- 14764316 TI - Value of clinical data and neuropsychological measures in probable Alzheimer's disease. AB - We examined retrospectively 60 probable Alzheimer's disease (AD) outpatients, 30 with early onset (EOP) and 30 with late onset (LOP), divided into two groups on the basis of illness duration (within 2 years (P<2) and over 2 years (P>2)), compared with 60 normal controls (NC). We employed a battery of neuropsychological tests including the mini mental state examination (MMSE) and our brief mental deterioration battery (BMDB), computerized psychomotor performance tests and staging of functional impairment. EOP were worse than LOP in verbal fluency and in functional impairment, being better only in Rey's long term verbal memory (RLT). P>2 were more compromised than P<2 in functional impairment, MMSE, personal and temporal orientation and RLT. Our BMDB showed the highest accuracy in classifying probably AD patients, whereas, MMSE had a high specificity but poor sensitivity as well as psychomotor performance tasks. In conclusion, AD patients with early onset, having a worse functional impairment, appear to be an eligible group to evaluate possible changes in response to antidementia treatment. PMID- 14764317 TI - Skin vessel reactivity tests in healthy middle-aged and elderly subjects: the influence of depolarizating current and serum lipids. AB - Ten healthy, non-smoking subjects without atopic constitution, (eight males and two females, mean age 67.3 +/- 2.5 years), participated in a study of vascular reactivity. The subjects were tested with respect to apolipoprotein E alleles. Three vasodilating substances were iontophoresed into the skin, acetylcholine chloride (ACh); isoprenaline sulfate (isoprenaline); and sodium nitroprusside (SNP). ACh and isoprenaline were delivered to the anode, while SNP was delivered to the cathode. Also, 0.9% sodium chloride (NaCl) was delivered to the cathode in order to assess the effect of the iontophoretic current itself. The resultant vasodilation was mapped by a newly developed laser Doppler perfusion imager (LDPI) and correlated with fasting concentrations of serum lipids and lipoproteins. Skin vessel reactivity test to ACh, isoprenaline and SNP, but not NaCl, showed statistically significant negative correlations to the ratio LDL cholesterol/HDL cholesterol (P<0.05). The skin vessel responses to endothelium dependent and possibly also to endothelium-independent substances are correlated with serum lipids and lipoproteins. It might be feasible to follow the effect of lipid-lowering strategies non-invasively with the aid of iontophoresis and laser Doppler perfusion imaging. The response to NaCl at the cathode might be due to a direct depolarizating effect on perivascular nerves and might thus be of interest when evaluating small nerve fiber dysfunction, for instance in diabetes mellitus. PMID- 14764318 TI - Posture-related tachycardia in older patients with hyponatremia. AB - Hyponatremia (HN) is the commonest electrolyte abnormality in elderly patients. Its etiology in this setting is poorly understood. In this study, the authors aim to compare the hemodynamic and hormonal responses of a group of older patients with a predisposition to HN with a group of age-matched controls. We assessed hemodynamic and hormonal responses to postural challenge in 15 patients over age 65 with serum sodium concentrations of less than 130 mM (mean 128.7 mM) and 15 age-matched controls with normal sodium concentrations. Patients remained recumbent for 1 h and stood for the second. Blood was drawn at baseline and at 15 min intervals. Blood pressure (BP) and pulse rates (PR) were monitored electronically. Plasma arginine vasopressin (AVP), atrial natriuretic peptide (ANP), renin and aldosterone were determined periodically during the study period. No difference in BP between groups was noted. PR increased significantly in the HN group only within 3 min of standing (from 71 +/- 4 to 86 +/- 5, P<0.01) and remained significantly higher than controls until 90 min (87 +/- 5 vs. 69 +/- 4, P<0.01). While plasma AVP levels increased significantly following 30 min standing and remained elevated for both HN and control groups, it did not differ significantly between the two. Baseline plasma ANP levels were significantly higher in HN patients compared with controls and remained significantly higher (P<0.05) throughout the study. There was no significant difference in plasma renin or aldosterone levels between groups during the study period. We have demonstrated differing autonomic and hormonal responses to orthostatic challenge between HN patients and age-matched controls. Water retention due to the syndrome of inappropriate anti-diuretic hormone secretion (with reset osmostat) may lead to raised ANP levels in this older cohort of patients. Further physiological studies are required to clarify the precise mechanism of these varying responses. PMID- 14764319 TI - 'What's his name?' A comparison of elderly participants' and undergraduate students' misnamings. AB - This study examined the effects of age on the types of errors produced when recalling names of faces. The types of errors included confusions (errors within the target set), intrusions (errors outside the target set), errors phonologically similar to the target, errors not phonologically similar to the target, and errors containing the same number of syllables as the target name. Participants included 49 elderly adults (57-85 years) and 48 undergraduate students (18-44 years). Age group had a significant effect on the number of name errors produced (n=681 for elderly and n=422 for undergraduates). Elderly participants produced more confusions than their younger counterparts; however, younger participants produced significantly more intrusions. The age groups also differed in their production of error names that were phonologically similar to the target name. The elderly participants produced more errors that were not phonologically similar to the target than the young adults. The results are discussed with regard to theories of name-face association and tip-of-the-tongue phenomena. PMID- 14764321 TI - What is lipofuscin? Defining characteristics and differentiation from other autofluorescent lysosomal storage bodies. AB - Lipofuscins, also known as age-pigments, have three defining characteristics: (1) they consist of intracellular secondary lysosomes; (2) they have a yellow autofluorescent emission when excited by near ultraviolet or blue light; and (3) they accumulate during normal senescence. Lysosomal storage bodies with similar fluorescence properties accumulate in various cell types as a result of specific pathological conditions or experimental manipulations. As a class, the latter are often referred to as ceroid pigments. In general, the mechanisms involved in the formation of ceroid pigments cannot be assumed to be closely similar to those involved in lipofuscin formation. In fact, the mechanisms of formation almost certainly differ, not only between lipofuscins and ceroids, but also among different lipofuscins and different ceroids. Presently, the most detailed knowledge about the mechanisms involved in lipofuscin formation come from studies on the retinal pigment epithelium (RPE) of the eye. These studies indicate that at least the autofluorescent constituents of RPE lipofuscin are generated from derivatives of vitamin A that occur in the retina. Oxidative stress to the retina appears to promote the formation of these RPE fluorophores. Whether similar mechanisms are involved in the formation of the lipofuscins that occur in other tissues remains to be determined. The mechanisms involved in RPE lipofuscin fluorophore formation are closely related to metabolic pathways that are specific to the retina. Thus, it appears likely that the mechanisms by which lipofuscins form in other tissues differ fundamentally from those that underlie RPE lipofuscin formation. PMID- 14764322 TI - Ceroid accumulation in rat kidneys caused by uptake of ferric nitrilotriacetate. AB - Ceroid accumulation in kidneys caused by lipid peroxidation in vivo was studied during intraperitoneal (i.p.) injections of ferric nitrilotriacetate (Fe3+-NTA) solution, which is a weak iron chelate compound and generates free radicals in the presence of oxygen. Fe3+-NTA solution (0.5-0.75 mg/100 g body weight) was injected into rats (n=6) for 4 days a week. After 2 weeks of Fe3+-NTA loading, the rats were not treated for 1 month as a rest interval, and then the iron loading was repeated for 35 weeks including the rest intervals. Two rats in each group, Fe3+-NTA loading and control, were sacrificed at 19 and 40 weeks of age, and portions of a kidney and the liver of each rat were fixed in 10% formalin buffered to pH 7.0 for the histological studies. We found ceroid accumulates in greater amounts and more quickly within macrophages and epithelial cells of proximal convoluted tubules of the kidney and Kupffer cells of the liver in iron loaded rats. PMID- 14764323 TI - Differential lectin histochemical studies on lipofuscin (age-pigment) and on selected ceroid pigments. AB - The persistent indiscriminate use of the term lipofuscin for the pigments encountered in pathological conditions, and which should be most properly termed ceroid pigments, is still creating unnecessary conceptual and nomenclature problems, and a great deal of confusion. While both the age-dependent lipofuscin and the pathologically formed ceroid pigments have somewhat similar physical and histochemical properties, sufficient differences to properly identify these two types of pigments are presented in this communication. In addition, because little is known on the saccharide components of lipofuscin and ceroid pigments in situ, we have in recent years explored the lectin binding characteristics of lipofuscin in human and rats, as well as in diverse ceroid pigments experimentally induced in rats. Our lectin histochemical results showed qualitative and quantitative differences in the saccharide composition between human cerebral neurolipofuscin and the intra and extracellular ceroid pigment of human atheromas, as well as, between rat lipofuscin and the ceroid pigments induced in these animals. PMID- 14764324 TI - The analytical approach to the nature of lipofuscin (age pigment). AB - Analytical studies of three lipopigments show that much can be achieved. Lipopigment from ovine ceroid-lipofuscinosis is composed of discrete protein and lipid molecules in orderly arrays and lipid peroxidation is not involved in its formation. Subunit c of mitochondrial ATP synthase accounts for approximately 50% of accumulated material and is specific to the disease process in this and other forms of the disease. Lipofuscin from bovine heart was mostly soluble and also contained discrete proteins, lipids and metals. Equine thyroid lipofuscin was less soluble but also had a relatively high protein content, probably derived from thyroglobulin. Although sugar could not be measured quantitatively, staining reactions and elemental analyses suggested it could also be a significant component. Some may be present as derivatives in the form of advanced glycation products. It is proposed that protein, the dominant molecular species present, is the important constituent in lipofuscinogenesis rather than lipid peroxidation. Whereas this latter may play some part in the maturation of lipofuscin, this has not been shown experimentally and is not likely to be the initiating mechanism. PMID- 14764325 TI - Sequential histochemical studies of neuronal lipofuscin in human cerebral cortex from the first to the ninth decade of life. AB - The typical and most consistent physico-histochemical properties of lipofuscin granules, such as autofluorescence, sudanophilia, acid-fastness, PAS-reactivity, and lectin reactivities for diverse saccharide moieties have been generally detected in tissue specimens of old humans and animals. The purpose of this study was, therefore, to explore possible sequential variations of each of these properties in cortical neurons of the left cerebral temporo-parietal areas from individuals dying from the first to the ninth decade. Autofluorescence was studied with an ad hoc equipped microscope, sudanophilia was evaluated by Oil-red O (ORO) staining, acid-fastness by long Ziehl-Nielsen reagent, PAS reactivity by the periodic-acid-Schiff reagent before and after diastase treatment, and the saccharide moieties by the use of a commercial kit of seven different biotinylated lectins. In the specimen from a 5-year-old child, lipofuscin granules were detected in less than 5% of the cortical neurons, but these granules already showed golden-yellow autofluorescence, sudanophilia, acid fastness and PAS-reactivity. From the second to the ninth decade of life, perikaryal lipofuscin granules were found in practically all cortical neurons with apparent agewise increases in the intensity of sudanophilia and PAS reactivity, but with variable acid-fastness expression. Surprisingly, however, no saccharide residues were detected by lectin histochemistry before the fifth decade of life. First detected saccharide was mannose in specimens from the fifth decade of life, and at later decades acetyl galactosamine, sialic acid and lactose were also found. Although, the reasons for the absence of lipofuscin affinity for the seven lectins used in this study in the cortical neurons of young and middle-aged individuals are presently unknown, these unexpected findings suggested important evolutionary changes of biogenesis and composition of the age-pigment. PMID- 14764326 TI - A flow-cytometric method for quantification of neurolipofuscin and comparison with existing histological and biochemical approaches. AB - The ability to measure lipofuscin accumulation accurately is essential for understanding its role in physiological ageing and human disease, and for its recent use as an ecological tool for age determination. Existing quantification methods are problematic. In situ histological measurement by microscopy can be very precise but is labour intensive. Spectrofluorimetric measurement of whole lipid extracts is rapid but not sufficiently specific. A recent HPLC assay for the retinal pigment epithelium lipofuscin fluorophore, A2-E, is potentially both precise and rapid but not applicable to lipofuscin in other tissues, or from fixed samples. In this study, I explore the use of flow cytometry or fluorescence activated cell sorting (FACS) for specific quantification of lipofuscin granules in formalin-fixed CNS homogenates from lobsters (Homarus gammarus). Free neurolipofuscin granules were discriminated in FACS samples by their size distribution (forward scatter), distinctive orange autofluorescence (FL3) and refractive internal structure (side scatter). A quantitative neurolipofuscin index was developed, which was highly correlated with the microscopically measured neurolipofuscin concentration in the same tissue. Sample-processing rate was at least an order of magnitude greater for FACS than for quantitative microscopy but the latter yielded a much more precise estimate of neurolipofuscin concentration. While the FACS approach may be ideal where rapid handling and only semiquantitative results are required, loss of precision will preclude use in many ecological studies where the highest available resolution is needed. Further refinements to the FACS approach are possible but advanced histological methods for neurolipofuscin quantification remain the most reliable at this time. PMID- 14764327 TI - Quantifying dense bodies and lipofuscin during aging: a morphologist's perspective. AB - Secondary lysosomes, residual or dense bodies containing lipofuscin or age pigment accumulate in post-mitotic and inter-mitotic cells during aging. The consensus is that the accumulation of this auto-fluorescent material is an index of cellular senescence. Biochemical and morphological studies have independently demonstrated marked age-related increases in the cell and tissue contents of lipofuscin. Most morphological studies on aging have been qualitative, have included only two or three age groups and have not yielded data that are easily correlated with biochemical analyses. One of the best documented age-related changes in hepatocytes and cardiac myocytes is the accumulation of dense bodies and lipofuscin inclusions. Independent stereologic studies reported two- to eightfold age-related increases in the dense body volume fraction of rat hepatocytes. Furthermore, we reported a fourfold increase in the dense body volume fraction of cardiac myocytes in rats between 6 and 30 months of age. These and other studies confirm the use of quantitative morphology to estimate the increases in dense body and lipofuscin inclusions as indices of age. Whether or not the accumulated lipofuscin compromises cell functions in senescent animals has not been adequately addressed. On the one hand, there is little evidence that several-fold increases in this subcellular compartment impair the functional capacities of either hepatocytes or cardiac myocytes. On the other hand, the age related accumulation of immunoprecipitable, but catalytically inactive, lysosomal enzymes in both liver and heart muscle may be a reflection of increased lipofuscin deposits in the dense bodies. PMID- 14764328 TI - Morphometry of age pigment (lipofuscin) and of ceroid pigment deposits associated with vitamin E deficiency. AB - Consistent amounts of lipofuscin and of ceroid pigment associated with vitamin E deficiency are reported to represent morphological correlates of aging and increased oxidative stress. A reliable quantification of these yellow autofluorescent deposits is of critical biological significance, thus we carried out a computer-assisted morphometric study on the accumulation of lipofuscin in physiological aging and of ceroid pigment in vitamin E deficiency, respectively. The total area and the size distribution of lipofuscin or ceroid pigment deposits were measured in CA3 hippocampal pyramidal neurons of 6-, 12-, 18- and 25-month old rats, as well as in vitamin E deficient animals of 18 months of age. An increase in the mean total area of lipofuscin and ceroid pigment was found in aging and in vitamin E deficiency. In both conditions, the proportion of large discrete deposits also increased. The similarity of changes observed in old and adult vitamin E deficient animals suggests that the underlying processes initiated by the absence of alpha-tocopherol from the diet of adult rats and in physiological aging may share some common mechanisms. PMID- 14764329 TI - Morphometric investigations of the mitochondrial damage in ceroid lipopigment accumulation due to vitamin E deficiency. AB - Numeric (Nv) and volume (Vv) densities, as well as the average size (skeleton: Sk) of synaptic mitochondria from adult, normally fed and adult, vitamin E deficient animals (11 months of age) were semiautomatically measured by computer assisted morphometry in the cerebellar granular layer. Nv, Vv and the average mitochondrial volume (V) were measured on perikaryal Purkinje cell organelles preferentially stained for succinic dehydrogenase (SDH) activity. Adult vitamin E deficient animals showed a significant decrease of Nv, a significant increase of Sk and an unchanged value of Vv. While in adult normally fed animals the mitochondria of increased size (Sk>5 microm) were 5.3%, in the adult vitamin E deficient rats this fraction accounted for 25.5%. In Purkinje cell perikarya, vitamin E deficiency resulted in a significant decrease of Vv, Nv and V, as well as a steeper reduction of the percentage of SDH-positive mitochondria of larger size. Taken together, these findings document that vitamin E deficiency is responsible of mitochondrial morphometric alterations in adult rats. Structurally deteriorated mitochondria are reported to play a role in producing increased amounts of free radicals, which can facilitate the accumulation of ceroid pigment. PMID- 14764330 TI - Promise and problems in relating cellular senescence in vitro to aging in vivo. AB - According to the 'Hayflick limit', human fetal fibroblasts have a uniform, limited replicative lifespan of about 50 population doublings in cell culture. This concept was extrapolated to diverse cells in the body. It seemed to decrease with the age of the cell donor and, as a form of cell senescence, was thought to underlie the aging process. More discriminating analysis, however, showed that the fibroblasts decayed in a stochastic manner from the time of their explantation, at a rate that increased with the number of population doublings in culture. There was no consistent relation to the age of the donor. Despite the contradictory evidence, the original version of the Hayflick limit retained its general acceptance. Cell senescence was attributed to the absence of telomerase in the fibroblasts, which resulted in shortening of telomeres at each division until they fell below a critical length needed for further division. However, it is well established that stem cells in renewing tissues undergo many more than 50 divisions in a lifetime, without apparent senescence. Contrary to early findings of no telomerase in most tissues, their stem cells retain telomerase and presumably telomere length despite many divisions in vivo. Massive accumulation of lipofuscin granules occurs under stress in long term crowded cultures, but the granules dissipate on subculture or neoplastic transformation. The overall results indicate a critical disjunction between cell senescence in vitro and aging in vivo. By contrast, cell culture has been useful in showing a need for telomere capping in maintaining cell stability and viability. It may also provide information about the biochemical mechanism of lipofuscin production. PMID- 14764331 TI - Role of environmental temperature in aging and longevity: insights from neurolipofuscin. AB - The available evidence for thermal modulation of neurolipofuscin deposition in poikilotherms is reviewed here and additional data are contributed. Mainly decapod crustacean models are employed and neurolipofuscin is treated as an index of physiological aging. In all cases, neurolipofuscin accumulation rate is positively correlated with environmental temperature but there appears to be lowered sensitivity in the thermal mid-range, an 'optimum' temperature for neurolipofuscin accumulation and possibly age-associated variation. The geographical position of the population within the species' thermal range may determine sensitivity of the response. There is seasonal oscillation of neurolipofuscin accumulation rate, providing preliminary evidence for neurolipofuscin turnover with net loss in winter. Spatial and temporal thermal variations of similar magnitude appear to have comparable effects on neurolipofuscin accumulation rate. Such effects may be extreme, suggesting important implications for physiological aging even in homeotherms. Inter specific comparisons indicate that species-specific neurolipofuscin accumulation rates are positively correlated with habitat temperature and inversely correlated with maximum lifespan and age at maturity. These findings help explain some well known bioclimatic trends in maturation- and maximum body size, such as Bergmann's rule. They also highlight the fact that global warming is likely to cause significant changes in life history parameters, population dynamics and responses to exploitation for many species. PMID- 14764332 TI - Potential reversibility of lipofuscin accumulation. AB - It is well established that the lipofuscin content of many post-mitotic cell types increases progressively during normal senescence. This age-related accumulation of lipofuscin may occur either: (1) because lipofuscin, once formed, is never degraded or eliminated from cells; or (2) because, despite turnover of this pigment, the rate of lipofuscin formation exceeds the rate at which it is eliminated. Little research has been performed to distinguish between these possibilities. Several studies suggest that lipofuscin may be turned over. However, definitive demonstrations that such turnover does occur under normal circumstances are lacking. It is possible to specifically halt new lipofuscin formation in the retinal pigment epithelium (RPE) of animals at an age when significant amounts of lipofuscin have accumulated in these cells. By monitoring the RPE for possible decreases in lipofuscin content after new pigment formation has been halted, it should be possible to determine whether previously formed lipofuscin can be eliminated from the RPE. PMID- 14764333 TI - Dietary factors in lipofuscinogenesis and ceroidogenesis. AB - The presence of ceroid pigments in human and animal tissues is associated with numerous pathological conditions in which the main pathogenic factor is the primary or secondary deficiency of vitamin E or imbalances between anti- and pro oxidants. That oxidative stress, particularly through its consequent lipid peroxidation, plays a capital role in the genesis of ceroid pigments, is supported by numerous in vitro and in vivo studies. Discussed in this presentation are two examples of oxidative stress on ceroidogenesis, namely the in vivo rat model of dietary hepatic necrosis, and the in vitro formation of ceroid pigments by the aerobic incubation of unsaturated fat and blood cells. Although it is widely believed that the progressive accumulation of lipofuscin is also a marker of oxidative stress, and that this pigment can be modulated by the dietary anti- and pro-oxidant factors, the evidence for these related notions is highly questionable. Some years ago, this controversial problem was reexplored in our laboratories by a series of studies in Wistar male rats, and the results indicated that neither the type of dietary fat, nor the pharmacological amounts of vitamin E significantly influenced the amounts of lipofuscin in cerebral neurons, cerebellar Purkinje cells, hepatocytes or cardiac myocytes. It was also found that the indices of lipid peroxidation determined in this study (production of malonaldehyde, and detection of conjugated dienes) did not correlate with the progressive accumulation of lipofuscin with age. All these results strongly suggest that the presence and cellular accumulation of lipofuscin can hardly be considered a marker of oxidative stress. PMID- 14764334 TI - The biological waste product formation in the light of the membrane hypothesis of aging. AB - The membrane hypothesis of aging (MHA) explains the biological waste product (lipofuscin) formation as a disbalance between the rates of protein synthesis and damage, as well as of elimination of the damaged components. Although, this concept has not been refuted on the basis of any experimental evidence, it has neither been widely accepted. During the last decade the general interest has turned toward the molecular genetics so intensely, that research aimed at clarifying cell biological mechanisms became so to say hibernated. Nowadays it is being recognized more and more that after the complete description of the human genetic code, attention has to be dedicated again to the cellular mechanisms explaining the function of the gene products (proteins). In this context, our experimental findings described during the recent years may become again the subject of interest. We have shown that the in vivo inhibition of the lysosomal thiol-proteinase functions by sublethal doses of leupeptin in young, adult and old mice results in a considerable increase (about 30%) of the immobile fraction of membrane proteins in hepatocyte plasma membrane, meanwhile the lateral diffusion constant of the still mobile membrane proteins increased. These observations were interpreted as signs of a general slowing down of protein turnover in the plasma membrane, just by inhibiting the elimination mechanisms in the lysosomes. This paper will discuss the theoretical conclusions and significance of these findings for the biological waste product formation, as a basic cell biological function. PMID- 14764335 TI - The origin of fluorescence in the neuronal ceroid lipofuscinoses (Batten disease) and neuron cultures from affected sheep for studies of neurodegeneration. AB - Lipofuscin and ceroid are usually held responsible for impaired cellular performance, via oxidative damage and the irreversible accumulation of fluorescent products of lipid peroxidation. The neuronal ceroid lipofuscinoses (NCLs, Batten disease) are inherited neurodegenerative diseases characterized by intracellular accumulation of fluorescent lipofuscin-like bodies. However these bodies are lysosomes packed with a particular protein, subunit c of mitochondrial ATP synthase; not the result of oxidative damage. No individual storage body component was fluorescent nor were solutions of total storage bodies. UV-vis spectra confirmed the lack of a fluorophor. Crystals of non-fluorescent albumin and reconstituted storage bodies were fluorescent in glycerol suspensions. This fluorescence is probably caused by interference of light reflected from the protein array, as is often observed in protein crystals. Other lipofuscins may be secondary lysosomes with a high protein content and the source of fluorescence the same. The neurodegeneration associated with lipofuscin accumulation may be caused by that accumulation, or may be a separate manifestation of aging. Neuronal cell cultures offer a way to study these processes. Subunit c accumulation has been observed in cerebral bipolar neurons cultured from 90 day NCL affected sheep foetuses. Neurons from different parts of the brain behave differently. Normal 108 day cerebellar granule neurons migrated into clumps when cultured with tri-iodothyronine, but affected cerebellar neurons did not, nor did normal or affected cerebral neurons. PMID- 14764336 TI - Potential role of retinal pigment epithelial lipofuscin accumulation in age related macular degeneration. AB - Age-related macular degeneration (AMD) is a leading cause of severe visual impairment in developed countries. The vision loss associated with AMD is the result of degenerative changes in the central region of the retina called the macula. Maintenance of normal structure and function of the macular retina, and of the remainder of the retina as well, is critically dependent on the supporting role of the adjacent retinal pigment epithelium (RPE). Impairment of normal RPE functions is known to result in retinal degeneration and loss of visual function. Thus, it has been hypothesized that the retinal degeneration that characterizes AMD is secondary to age-related deterioration in RPE support functions. Like many other postmitotic cell types, the RPE accumulates autofluorescent lysosomal storage bodies (lipofuscin) during senescence. In human eyes, lipofuscin comes to occupy a substantial fraction of the RPE cytoplasmic volume in the elderly. Does this lipofuscin accumulation contribute to the development of AMD? This question is a specific case of the broader question of whether lipofuscin accumulation in general is detrimental to cells. Unfortunately, definitive data do not exist to allow these questions to be answered. Although a correlation between RPE lipofuscin content and AMD has been reported, a cause-and-effect relationship between RPE lipofuscin accumulation and the development of this disease has not been established. It has been reported that a mutation in a gene encoding a photoreceptor-specific protein results in massive RPE lipofuscin accumulation and early-onset macular degeneration. However, again the accelerated RPE lipofuscin accumulation has not been shown to be the cause of the accompanying macular degeneration. The lack of a definitive link between RPE lipofuscin accumulation and AMD illustrates one of the biggest challenges remaining in lipofuscin research-determining whether lipofuscin accumulation per se has an impact on cell function. PMID- 14764337 TI - Age-related antioxidant capacity of the vitreous and its possible relationship with simultaneous changes in photoreceptors, retinal pigment epithelium and Bruchs' membrane in human donors' eyes. AB - It is well established that the retina undergoes several decremental functional and structural changes with age, and it has been suggested that most of these age related changes may be mainly due to oxidative stress through light-induced mechanisms. While the cornea and the lens absorb part of the light radiation, the vitreous appears to be the main site of protection against radiation damage to the retina. Thus the purpose of this study was to explore possible interrelationships between the oxidative capacity of the vitreous and the number of photoreceptors, the fluorescent intensity of the retina, and the thickness of the Bruch's membrane in eyes from donors dying from the third to the eighth decade of life. The results showed that the total reactive antioxidant potential of the vitreous significantly decreased with the age of the donor's eyes, and that this decrease was directly correlated with decreases in the photoreceptors and inversely correlated with increases in the autofluorescent intensity of the retina and the thickness of the Bruch's membrane. Although these correlations did not prove a cause-effect relationship, these findings may provide new insights for the understanding and possible treatment of the age-dependent changes of the human eyes. PMID- 14764338 TI - Age related prolactin secretion in men after fentanyl anaesthesia. AB - The aim of this study was to investigate the role of age in the hormonal response to opiate anaesthetic fentanyl. In 90 patients undergoing aortocoronary bypass, 59.6 +/- 9.2 years mean age, 35-81 age range, prolactin (PRL), thyroid stimulating hormone (TSH), follicle stimulating hormone (FSH), luteinizing hormone (LH), human growth hormone (HGH), insulin-like growth factor I (IGF I), glucagon and insulin were measured in venous blood samples drawn from fasting patients immediately before, at 8 h in the morning, and 60 min after the induction of anaesthesia with 30 microg/kg intravenous fentanyl bolus, 30 min after a second 7 microg/kg fentanyl bolus. Results showed a higher 60 min PRL peak in older, >65 years, in respect to younger, < or =50 years, patients (57.6 +/- 23.3 vs. 40.6 +/- 13.8 microg/l, P<0.005), with a significant upward trend with age across the entire age span (r=0.32; P<0.002), while no difference by age was found for the basal concentrations. No differences were found between the respective basal and 60 min concentrations for the other hormones investigated. As expected, differences by age were found for FSH, higher in >65 and in 51-65 year-olds than in younger patients (for the basal values, respectively, P<0.02 and P<0.05); IGF I was lower in >65 in respect to < or =50 (P<0.02) and to 51-65 year-old patients (P<0.05), with a significant negative correlation with age (r= 0.33; P<0.005). The study shows an age related increase of PRL concentrations after fentanyl administration. It may be due to the reduction of the hypothalamic dopaminergic tone with aging. IGF I levels have been confirmed to be inversely correlated with age. PMID- 14764339 TI - Quality of life in dependent older adults living at home. AB - The purpose of medical interventions today is to favor the duration of life and to assure its quality. For a proper evaluation of the quality of life (QoL) of the elderly, it is important to assess not only the health status, but also psychological, functional and existential domains. Up to now, QoL of the dependent older living at home does not seem to have received sufficient attention. This study is a population based, cross-sectional health survey, and tries to give a picture of the QoL conditions of a group of dependent elderly living at home, using a specifically designed structured interview. In addition to this interview the Activities of Daily Living (ADL) Index and the Zung rating scale for Anxiety and Depression (AD) were administered to a total of 167 elderly (60 males and 107 females). The most interesting results of the study are the following: (1) the definition of dependence is often worsen by cultural prejudices; (2) psychopathological factors show a deep negative effect on 'attitudes towards life' of the dependent elderly; (3) QoL of the dependent elderly people in this sample is mainly negatively influenced by the degree of depression. We believe that the 'QoL-oriented' therapeutic strategy should consider more articulated and multidisciplinary geriatric and psychosocial interventions in this population. PMID- 14764340 TI - Smoking habits and attitudes of age concern volunteers. AB - As the population is ageing, health promotion is becoming increasingly important to prevent disease and disability. Cigarette smoking is strongly associated with excess mortality and smoking cessation even after the age of 65 years, improves health and lowers mortality. The voluntary sector is an under-utilised resource for health promotion to older people. Age Concern Cymru is the leading voluntary organisation in Wales involved in promoting 'healthy ageing'. The aim of this study was to determine the smoking habits and attitudes of older Age Concern volunteers. A questionnaire enquiring about smoking habits and attitudes was circulated to all volunteers aged 65 years and over attending age concern meetings in Wales between July and September 1999. Of 375 respondents (93% response rate) 16% were current-smokers, 58% ex-smokers and 26% life-long non smokers. A significantly greater proportion of women were non-smokers. Significantly more ex-smokers (90%) and non-smokers (93%) compared with current smokers (72%) believed that smoking causes heart disease (P<0.002). More ex smokers (91%) and non-smokers (94%) than current-smokers (80%) believed smoking causes lung cancer (P<0.05). In conclusion, the majority of volunteers are currently non-smokers. Over 90% of ex-smokers and non-smokers believed in the deleterious effects of smoking on health. These volunteers could be utilised for smoking-cessation campaigns. PMID- 14764341 TI - Urinary incontinence in the elderly: relation to cognitive and motor function. AB - Urinary incontinence is a common problem in older subjects, very often wrongfully accepted as a normal part of the aging process. A total of 520 subjects (208 males and 312 females; mean age 74.8 +/- 11.8 years), from both private- and nursing-home dwelling populations, were included in this study aimed to estimate the incidence of urinary incontinence and identify factors associated with condition, in aged subjects. The incidence and type of urinary incontinence (stress, urge or mixed incontinence) were assessed by structured questionnaires and diagnosis was confirmed by a seven-day consecutive voiding diary. Assessment of physical, cognitive and emotional functions was performed on each subject using the Mini Mental State Examination (MMSE), Instrumental Activities of Daily Living Scale (IADL), Tinetti Scale (gait), Tinetti Scale (balance) and Geriatric Depression Scale (GDS) instruments. In the total population sample the incidence of urinary incontinence was 47.9%. The incontinence cases were classified, according to the different types, as: stress incontinence (males: 3.4%; females: 8.7%; males+females: 6.5%); urge incontinence (males: 27.4%; females: 31.4%; males+females: 29.8%); mixed incontinence (males: 20.2%; females: 5.8%; males+females: 11.5%). In the total population sample, no significant relationship was found between age and prevalence of urinary incontinence. In the elderly female group, age significantly correlated in a direct manner with urge incontinence (P<0.01) and inversely with stress incontinence (P<0.001). Only in the male sex group age significantly correlated with mixed incontinence (P<0.005). Multiple linear regression analysis showed that the dependent variable 'incontinence' could be predicted by MMSE (P<0.001) in the male sex group and by the Tinetti Scale (gait) (P<0.001) in the female sex group. PMID- 14764342 TI - Prescribing behavior for the elderly in the United Arab Emirates: psychotropic medication use remains low despite rising overall appropriate and inappropriate medication use. AB - Inappropriate prescribing, especially for psychotropic agents, is a common and significant cause of morbidity in older people. This cross sectional survey was conducted in the United Arab Emirates (UAE), a country with a rapidly developing economy. Prescribing behavior for people aged 65+ years acutely admitted via a university teaching hospital was examined for 1994 and 1999. All 474 patients (which resulted in 627 acute admissions; 194 in 1994 and 433 in 1999) were included. The patients had a mean age of 74.1 +/- 7.6, mean annual admission rate of 1.3 +/- 0.8 and a female:male ratio of 0.46 in 1994 rising to 0.73 in 1999 (P=0.04). Pre-admission use of five or more medications as recorded in the admission notes increased from 12% in 1994 to 23% in 1999 (P=0.001), while on discharge rose from 26 to 45% (P<0.001). There was a rise in low dose aspirin use, an indicator of appropriate prescribing, both pre-hospital (13-21%, P=0.03) and on discharge (19-29%, P=0.007). There was also a significant rise in pre hospital inappropriate prescribing from 5 to 13% of patients demonstrating at least 1 of 144 inappropriate medications or combinations looked for (P=0.002) and at discharge from 9 to 19% (P=0.001). The rate of psychotropic medication usage (pre-hospital 0.1 per person: at discharge 0.25) was low compared to western countries and showed no significant change over time. These findings show rises in both appropriate and inappropriate prescribing with the exception of psychotropic medications during a 5-year period, which corresponded to rapid development in the health care system. PMID- 14764343 TI - Spironolactone therapy in older patients--the impact of renal dysfunction. AB - Low dose spironolactone reduces the risk of death from heart failure. We examined the effects of spironolactone on potassium homeostasis in a cohort of elderly patients with congestive heart failure (CHF). Eighteen patients >70 years, mean 80.5 (+/- SD 6.3) with New York Heart Association CHF Grade II-IV were enrolled. All patients were commenced on 25 mg spironolactone daily. The dose was reduced to 12.5 mg daily when hyperkalemia (potassium>5.0) occurred. A serum creatinine of >150 micromol/l was defined as indicating renal impairment (RI). Blood pressure, pulse rate, urea, creatinine, Na+ and K+ were measured at baseline, day 2-5, day 28 and more often if clinically indicated. Nine of those recruited had RI. Baseline serum potassium was significantly higher in those with RI, mean 4.56 (+/- 0.30) vs. 4.04 (+/- 0.30) mmol/l (P<0.01). Six patients with RI developed hyperkalemia versus one of those with serum creatinine <150 micromol/l (P<0.05). Serum K+ returned to normal in all patients when the dose of spironolactone was reduced to 12.5 mg daily with one exception in whom the medication was withdrawn. When spironolactone is prescribed to older patients with CHF, hyperkalemia appears more likely in those with RI. Halving the dose to 12.5 mg daily results in normalisation of serum potassium. Older patients commencing spironolactone therapy should have serum potassium monitored frequently, particularly in the presence of RI. PMID- 14764344 TI - On the antiatherogenity of calcium channel blockers: studies in proliferating vascular smooth muscle cells on age sensitivity, dose dependent inhibitory effect, and time of action. AB - We study the effect of diltiazem on cultured vascular smooth muscle cells from humans and rats, paying special attention to its activity in relation to the concentrations applied, incubation times after addition and the capacity to act against the mitogenic activity of insulin and insulin-like growth factor 1 (IGF 1). The mitotic activity was measured by means of bromodeoxyuridine DNA incorporation. Smooth muscle cells from old individuals showed a dose-dependent regression of the inhibitory level but not those from the young subjects, which showed a remarkable inhibition of mitosis at all concentrations tested. Around 8 h after addition, diltiazem inhibited cell proliferation at all concentrations tested. The inhibition exerted by 10(-7) M rapidly disappeared, reaching values higher than those initially registered and returning to basal rates after 72 h. The inhibition by 10(-6) and 10(-5) M remained after 30 and 72 h, respectively. Insulin (100 nM) or IGF-1 (1 nM) did not counteract the inhibitory effect of diltiazem (10(-5) M). Despite differences related to doses and age of cells, we conclude that diltiazem--as an L-type calcium channels blocker--is a potent inhibitor of vascular smooth muscle cell proliferation. PMID- 14764345 TI - Metalloproteinases (MMPs -2, -3) are involved in TGF-beta and IGF-1-induced bone defect healing in 20-month-old female rats. AB - Matrix metalloproteinases are important in the physiological and pathological degradation of extracellular matrix including that of bone and cartilage. The process of bone defect healing is associated with formation of cartilage callus and cancelous bone. With maturation and aging, the response of skeletal tissues to injury is limited. The ability of growth factors to enhance bone defect healing in aged rats was studied. Partial bone defects were induced in femurs of aged rats. A single dose of IGF-1, TGF-beta+IGF-1 or saline was inserted in the defect and bones were examined after 2 and 4 weeks. Morphology revealed that after 2 weeks of treatment with TGF-beta the defects were filled with mesenchyme like tissue and delicate bone trabeculae. Positive staining for metalloproteinase 2 (MMP-2) was shown at the sites of new bone formation. In defects treated with IGF-1 or TGF-beta+IGF-1 nodules of cartilage and fine bone trabeculae along with positive staining for both MMP-2 and MMP-3 were demonstrated in the healing defects. After 4 weeks radiology revealed mineralization in defects treated with TGF-beta and less pronounced mineralization after treatment with IGF-1, or with TGF-beta+IGF-1, whereas only partial healing of the defects was observed in control specimens. MMP-2 and MMP-3 were detected at sites of new bone formation after treatment with TGF-beta, IGF-1, and TGF-beta+IGF-1. It is concluded that TGF-beta and IGF-1 induced bone defect healing in aged rats. TGF-beta induced bone formation while IGF-1 induced cartilage and than bone formation via endochondral ossification. The localization of MMP-2 and MMP-3 in the healing defects reflected the synthesis of bone or cartilage matrices in the defect, reflecting the involvement of MMPs in the process of bone formation and endochondral ossification. The ability to induce bone defect healing in aging is of great clinical importance and understanding the involvement of MMPs in this process can contribute to future treatment with growth factors to enhance bone defect healing in 20-month-old female rats. PMID- 14764346 TI - Does the combination of a simultaneous subcapital fracture of humerus and hip fracture in elderly patients carry a prognostic value? AB - The most important factor to cause hip fractures in elderly is probably osteoporosis. Other factors are the increase in fall frequency and the protective response to trauma. Osteoporotic fractures occur most commonly, at the hip, vertebra, distal radius and proximal humerus. A combination of these is uncommon. Thirty-two women and six men treated between January 1990 and December 1999 for a combination of subcapital fracture of the humerus and hip fractures were evaluated retrospectively. The following parameters were reviewed: age, sex, pre fall function, use of drugs, chronic and acute comorbidity, circumstances of the fall, length of hospitalization, treatment procedure, complications and post hospitalization rehabilitation. Group I consisted of 15 patients aged 70-80 years and group II consisted of the remaining 23 patients, older than 80 years. In all 38 patients the simultaneous fractures were ipsilaterally. Hospital stay ranged from 7 to 17 days for the discharged 37 patients. Twenty-six of 28 patients, who were transferred to a rehabilitation center, returned to their previous activity of daily living (ADL). Among the nine remaining patients only five gained full recovery. A combination of fractures, occurs in the higher-age group, is quite traumatic to the patient and probably involves a greater impact force. In all patients it occurs in the ipsilateral side. In the elderly, even a minimal transmission to the osteoporotic hip can cause a fracture. The double trauma represents a better pre-morbid condition relative to patients in the same age group, thus it may serve as a prognostic indicator for success in rehabilitation. PMID- 14764347 TI - Changes in health, functional performance and activity predict changes in self rated health: a 10-year follow-up study in older people. AB - The purpose was to examine changes in self-rated health (SRH) in older people and associations between these changes and various self-reported and objectively measured indicators of health status, functional performance and activity at three time-points 5 years apart. Further, our aim was to examine whether SRH takes the form of a continuum. The study group comprised all the baseline 75-year old inhabitants of the City of Jyvaskyla, Finland (N=382). Four groups were formed according to change/stability in SRH: 'good-good', 'good-bad', 'bad-good' and 'bad-bad'. Cross-tabulation and one-way analysis of variance (ANOVA) was used to examine the cross-sectional differences, and GLM Repeated Measures to analyze the differences in changes in the determinants of SRH over time. Stability was more common than change in SRH, although in most participants there was a decline in most of the determinants of SRH. A systematic relation was found between the SRH groups and the indicators of health status, functional performance and physical and social activity, especially over the first 5-year period. A substantial decrease in these indicators was associated with a decline in SRH. Among older people change and stability in SRH over time systematically reflect health status, functional performance, and physical and social activity. The relatively high stability found in SRH indicates that with increasing age older people adapt to their worsening health conditions. SRH seems to form a continuum when a wide range of self-reported and objectively measured indicators on physical, psychological and social components of health are taken into account. PMID- 14764348 TI - Cognitive functioning in a population-based sample of very old non-demented and non-depressed persons: the impact of diabetes. AB - The main objectives of this study were to examine potential effects on cognitive functioning of diabetes and some major cardiovascular signs and diseases in very old age. Analyses were based on data from the Kungsholmen project and involved a population-based sample of non-demented and non-depressed persons aged 75-96 years. Diabetics (n=36) were compared with non-diabetic persons (n=356), and all participants were assessed with an extensive medical examination, specific diagnostic interviews, and laboratory blood analyses. Cognitive ability was indexed by means of the Mini-Mental State Examination (MMSE). The results showed that diabetics perform significantly worse on the MMSE than do non-diabetics, and that the MMSE-performance of elderly with cardiovascular disease (CVD) is significantly worse than in those without CVD. Controlling for cardiovascular signs and diseases did not alter the effects of diabetes. Examination of interaction terms, finally, indicated that, in the examined age range, the effects of diabetes do not vary across age, educational levels or variations in cardiovascular health. The results suggest that cognitive deficits associated with diabetes in very old age may be detected with the MMSE. PMID- 14764349 TI - Functional transitions and active life expectancy for older Japanese living in a community. AB - We examined functional transitions in both the activities of daily living (ADL), and instrumental activities of daily living (IADL), over a 1-year interval among older Japanese living in a community, then estimated their physically active life expectancy (PALE) and instrumentally active life expectancy (IALE). In 1992, all residents aged 65 or older who lived in Saku City, Nagano, Japan, were followed up over the 1-year interval. A self-administrated questionnaire, which involved age, sex, five ADL items, and five IADL items, was used for each survey in both 1992 and 1993. Of the baseline cohort (n=10,098), we received 9533 analyzable questionnaires at the follow-up survey in 1993. During the follow-up, 92 and 87% of subjects who were initially independent in ADL and IADL remained independent, respectively. PALE for men and women were estimated to be 16.0 and 18.9 at 65 years of age, respectively. IALE for men and for women were estimated to be 12.8 and 14.6 at the age of 65, respectively. Proportions of the PALE to total life expectancy (TLE) at any age did not greatly differ between men and women; however, men had a slightly larger proportion of IALE to TLE at any given age than women. PMID- 14764350 TI - Evaluating elderly patients with syncope. AB - Age-related physiologic changes together with high prevalence of chronic illness and the use of multiple medications predispose older adults to syncope. The causes of many of these patients' syncope are unexplained and most remained symptomatic. A thoughtful diagnostic strategy is essential to pinpoint the cause in each patient. The initial assessment includes a comprehensive medical history, preferably with an eye witness account, and a thorough physical examination. The decision for further diagnostic tests often depends on whether there is evidence of underlying structural heart disease. In the absence of heart disease, tilt table testing and the related autonomic function testing are usually most productive. Various cardiac studies will be more appropriate for those with suspected structural heart disease. A cause of syncope can only be concluded if there is a sufficiently strong correlation between syncopal symptoms and the detected abnormalities on investigations. A strategic evaluation of syncope of the elderly subjects should allow a correct diagnosis and appropriate management. PMID- 14764351 TI - Clinical brain proton magnetic resonance spectroscopy for management of Alzheimer's and sub-cortical ischemic vascular dementia in older people. AB - This study examined the clinical usefulness of magnetic resonance spectroscopy (MRS) performed using an automated single voxel technique at 1.0 T field strength in a district general hospital magnetic resonance (MR) scanner in the assessment of older people referred to a memory clinic with suspected dementia. Of 50 elderly subjects (M:F 20:30) examined and followed-up clinically over more than 2 years, 20 had clinical Alzheimer's disease (AD), 18 had clinical vascular dementia, six had mixed features and three were normal. Three normal volunteers were also studied. MRS was performed at the same time as structural magnetic resonance imaging (MRI), added <15 min to the examination and was well-tolerated in all patients studied. Patients with AD had significantly higher myoinositol/creatine (MI/Cr) ratios (mean +/- S.D.: 0.82 +/- 0.04) compared to those with vascular dementia (mean +/-S. D.: 0.71 +/- 0.07, P<0.00001) and normal subjects (mean +/- S.D.: 0.72 +/- 0.036, P<0.0002); there was little overlap between the AD and vascular groups. The mixed dementia group also had significantly higher MI/Cr ratios (mean +/- S.D.: 0.80 +/- 0.05) than vascular dementia (P<0.01) and normal (P<0.03) groups, but with considerable overlap. No significant differences were shown for N-acetyl aspartate or choline/creatine ratios between the different clinical groups. These data suggest that MI/Cr ratios can distinguish patients with AD from normal subjects and those with sub cortical ischemic vascular dementia and that MRS will be useful to clinicians managing persons with AD in a district general hospital setting. PMID- 14764352 TI - Surgical treatment of lumbar spinal stenosis in patients aged 65 years and older. AB - Spinal stenosis syndrome affects mainly patients at their 5th-6th decades of life. The main goals of surgical treatment in the elderly are to allow the individual to walk longer distances, maintain the activities of daily living (ADL) and social life. Our aim was to evaluate the results of surgical treatment for lumbar spinal stenosis in elderly patients. All patients over 65 years of age who underwent surgery due to spinal stenosis syndrome between 1990 and 1998 were evaluated. There were 29 males and 17 females aged between 65 and 90 years. The clinical presentation included low back pain (89%), intermittent claudication (100%) and neurological involvement (87%). The radiological examination showed a frequent narrowing at the level L4-L5 in 93.5% of the patients. The results of the surgery in a mean follow-up of 22 months were good to excellent in 80% of the patients, fair in 11%, and poor in 9%. An improvement in the intensity of pain and in walking distances was noted in 89 and 85% of the patients, respectively. Improvement was achieved in the level of daily activity and in social lives in 57 and 61%, respectively. Major and minor complication rates were 6.5 and 19.5%, respectively. No mortality was noted in this series. Eighty-seven percent of the patients were satisfied with the results of the surgery. We conclude that Surgery for spinal stenosis is a successful and relatively safe procedure, also for patients aged over 65, and should be considered as a treatment option for these patients. PMID- 14764353 TI - Clock drawing task, mini-mental state examination and cognitive-functional independence measure: relation to functional outcome of stroke patients. AB - The use of reliable and valid brief cognitive screening instrument for selecting the appropriate candidates for stroke rehabilitation is crucial. Clinicians often face the question which test should be preferred, that will best correlate with functional outcome. The objective of this study was to compare the clock drawing task with other cognitive tests used for the evaluation of discharge functional outcome in elderly stroke patients. We conducted a retrospective chart study including 151 consecutive patients, admitted for inpatient comprehensive rehabilitation following acute stroke. The clock drawing task (CDT), mini-mental state examination (MMSE) and the cognitive-functional independence measure (cognFIM) were used to assess the cognitive status. Functional status outcome was evaluated by the functional independence measure (FIM), using absolute and relative parameters of efficacy and efficiency. Correlation coefficients (Pearson correlation) between the three cognitive tests resulted in r-values ranging from 0.51 to 0.59 (P<0.001). All three tests correlated significantly with motor outcomes. MMSE did not confer additive value to CDT. It is concluded that CDT is similar to mini-mental and both are somewhat better than cognFIM with respect to the evaluation of functional status outcome following stroke. The correlations between the tests as well as the simplicity of administration favor the use of either CDT or MMSE in the initial assessment of elderly stroke patients. PMID- 14764354 TI - The impact of ADL status, dementia and body mass index on normal body temperature in elderly nursing home residents. AB - A subset of seniors might demonstrate a lower body temperature compared with younger subjects. However, data on normal body temperature in seniors are sparse. The aim of the study was to study normal body temperature with a view of predicting factors of low body temperature in non-febrile seniors. Elderly women (n=159) and 78 men, aged > or =65 years, living in community resident homes were included in the study. Data on chronic diseases and medication were collected from medical records. Tympanic and rectal temperature was measured twice daily; once at 7-9 AM and then at 6-8 PM. In addition, body mass index (BMI), activities of daily living (ADL) status, as well as details regarding dementia and malnutrition were recorded. The variation in tympanic and rectal temperatures ranged from 33.8 to 38.4 degrees C and 35.6 to 38.0 degrees C, respectively. ADL status, dementia and BMI were significantly related to lower and analgesic to higher tympanic temperature. Dementia was significantly related to lower rectal temperature. Therefore, dementia, BMI, ADL status and analgesic shall not be overlooked when assessing temperature in seniors. More research is needed to further clarify the influence of these predictive factors, as well as the impact of BMI and malnutrition. PMID- 14764355 TI - Survival curve modified through dietary restriction (DR) in male Donryu rats. AB - It is well known that dietary restriction (DR) can prolong the life of certain inbred strains of laboratory rodents. This prolongation can be usually shown in a survival curve in parallel to that of non-DR groups. On an outbred strain, Donryu, however, DR resulted in a unique pattern of survival curve. In DR group, the first three quarters of male rats survived longer in a form parallel to non DR, while the last quarter of them survived far longer in a quite different shape from controlled group. DR may select animals that are capable of longer survival within Donryu. The selection of a quarter animal indicates the parameter for selection may be regulated by a single recessive locus. PMID- 14764356 TI - The development of sleep in persons undergoing cataract surgery. AB - This study was undertaken in order to analyse sleep in a group of patients who were operated on for cataract. All patients (n=328) undergoing cataract surgery at the Department of Ophthalmology, Sundsvall Hospital during a 4-month period were asked to complete a questionnaire on the state and change of sleep and sleepiness 1 month after the operation. Twelve persons were unable or declined to participate. The response rate was 97.2%. The mean ages of the participating men and women were 74.5 and 76.3 years, respectively. Pre-operative visual acuity in the operated eye was 0.16 in men and 0.18 in women. After cataract extraction sleep was improved in 12.0% of the men and in 26.3% of the women. Nevertheless poor sleep 1 month post-operatively was reported by 29.3% of the men and 42.6% of the women (P<0.05). There was no age-related increase in sleep complaints. The results indicate that in elderly persons with cataract sleep is impaired, and that 1 month after cataract extraction improved sleep may be experienced. PMID- 14764357 TI - An expression of concern on the paper "Sleep and hypnotic use in relation to perceived somatic and mental health among the elderly" by Dr Ragnar Asplund (Archives of Gerontology and Geriatrics, 31, 2000). PMID- 14764358 TI - Hypersexuality in nursing care facilities--a descriptive study. AB - The continuance of sexual expression in the elderly as age advances is well recognized. Sexual disinhibition, however, in a restricted environment such as in nursing care facilities has received scant attention. We wish to describe eight patients residing in nursing care facilities who were seen because of their problematic sexually related behaviors. These behaviors include cuddling, touching of the genitals, sexual remarks propositioning, grabbing and groping, use of obscene language and masturbating without shame. In all instances concern emanated from members of the nursing staff. Other associated behaviors included aggression, agitation, and irritability amongst others. The computed tomography (CT) scan of the brain showed infarction in the frontal lobe (4), parietal lobe (1), and the caudate (1). One had severe Parkinson's Disease and one had severe dementia of the Alzheimer's disease. All ten patients had an organic basis for their symptoms. Sexually inappropriate behaviors remain highly controversial and labeling them as 'diseased' or an 'illness' may have enormous individual, cultural and medico-legal implications. The clinico-anatomical correlation are discussed. PMID- 14764359 TI - Superoxide dismutase, catalase and glutathione peroxidase activities in the liver of young and old mice: linear regression and correlation. AB - Activities of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GP) were determined in the liver of 15 young (3-5 months) and 15 old (23-26 months) female CBA mice. Although, the activities of all three enzymes decreased in aging, only the SOD decline was statistically significant (P<0.01). The pair wise correlation analysis exhibited an almost identical positive correlation between SOD and CAT in the liver of both young and old mice (r=0.57), whereas the correlative links between the enzymatic pairs of SOD-GP and CAT-GP were increased in aging. Close to zero in the young group (r=-0.08), the coefficient of correlation between SOD and GP became highly significant in the group of old mice (r=0.66; P<0.01). When the coefficients of pair wise and partial correlation were compared, practically no differences were found for the young mice, whereas all three partial coefficients were decreased in the group of old animals. Estimation of the linear regression between the enzymatic pairs revealed higher coefficients of regression and lower intercepts in the group of old mice. The results imply stronger correlative links between the antioxidant enzymes in the liver of old mice, compared with the same indices in the liver of young animals. PMID- 14764360 TI - Comparison of the reaction time, the Berg Scale and the ABC in non-fallers and fallers. AB - Postural sway, reaction time, the Berg Scale and the Activities-specific Balance Confidence Scale (ABC) were administered to 80 elderly subjects (40 fallers and 40 healthy non-fallers) in order to determine reliable predictor for those at risk of falling. Non-fallers showed significantly faster reaction times when compared with fallers experimental group. Postural sway results show a higher oscillation displacement in the Y axis (lateral) than the X axis (anterior posterior) and that fallers oscillate at a higher frequency than non-fallers. All variables were submitted to a multiple regression analysis. Only three of the four variables were found to be significant predictors of fall status. The variables in order of their inclusion are reaction time, Total Berg Balance Score and mean ABC. It was further determined that reaction time was indeed the best predictor of fall status in the present study. PMID- 14764361 TI - The prevalence of various arrhythmias in normotensive and hypertensive elderly patients. AB - The prevalence of the most frequent arrhythmias was studied in a geriatric day hospital. Patients older than 65 years have been considered, of them 118 were normotensive and 56 hypertensive subjects. Comparison of the two groups revealed no statistically significant difference either in the frequency of occurrence or in the type of arrhythmias. Dividing the hypertensive patients in the Lown's classes, only those of class 3 or over displayed a considerably higher frequency of arrhythmias than the normotensive subjects. The results demonstrate the age dependent increase of the prevalence of arrhythmias in general, and the increased occurrence of complex ventricular types in the hypertensive subjects. PMID- 14764362 TI - Urinary incontinence and related symptoms in older men and women studied longitudinally between 70 and 97 years of age. A population study. AB - The aim of this study was to assess the prevalence of urinary incontinence and other lower urinary tract symptoms among an aged population and follow this prevalence longitudinally. A random sample of 70-years-olds participated and the survivors have been reinvestigated longitudinally, between 70 and 97 years of age. The prevalence of urinary incontinence was 17% among men and 48% among women 70 years of age. No change in the total prevalence of urinary incontinence or stress incontinence was seen among those that were investigated at the age of 70, 75, 79, 81, 85, 88, 90, 92, 95 and 97. There was some increase in the prevalence of urge incontinence among men and women and some increase in the prevalence of weak stream among men. Urinary catheters were uncommon in this investigation, even among the oldest, and those with long-term catheter use had a high risk of dying. In this study, urinary incontinence was common among men and women 70 years of age. Those who participated in all examinations up to very high age had no increase in prevalence of total urinary incontinence or stress incontinence, but there was some increase with increasing age in the prevalence of urge incontinence and weak stream. These increases were, however, smaller than those found in cross-sectional studies, perhaps due to the fact that those who attended these investigations were healthy and active and, therefore, not afflicted as much, and when they were, they tended to seek help more often. Indwelling catheter use was uncommon in this group of elderly individuals and those who had a catheter had a high mortality, probably due to the fact that only the very ill get an indwelling catheter in Sweden today. PMID- 14764363 TI - Exposure to opioid analgesia in cognitively impaired and delirious elderly hip fracture patients. AB - The objectives of this study were to characterize patterns of opioid analgesia in elderly hip fracture patients, to investigate the possible differences in the treatment of cognitively impaired, delirious, or cognitively intact patients, and to study the factors that may affect the doses received by such patients. This retrospective study comprised 184 elderly patients with hip fractures undergoing surgical fixation. Data collection included age, sex, length of stay, type of fracture, cognitive status by mini-mental state examination, assessment of possible delirium by the confusion assessment method, type and doses of opioid received by these patients. We found that the amount of morphine equianalgesic dose differed significantly between demented and non-demented patients (7.5 +/- 1.8 vs. 14.1 +/- 4.9, P<0.001). Patients with cognitive decline or with delirium received only 53 and 34%, respectively, of the amount of opioid that was administered to cognitively intact patients. A significant association was observed between cognitive status, or delirium, and amount of opioid analgesia (P<0.001 and P=0.003, respectively). Other parameters such as age, length of stay and type of fracture, had no effect on the use of opioid analgesia. It is concluded that the management of pain in older persons with hip fracture surgery is suboptimal with regards to insufficient administration of opioid analgesia in demented and delirious patients. The adoption of a standardized protocol for pain control may help in reducing the extent of this problem. PMID- 14764364 TI - Meperidine analgesia and delirium in aged hip fracture patients. AB - Delirium is quite frequent in elderly patients who sustain hip fractures. The use of Meperidine by physicians, unaware of the possible emergence of delirium in elderly patients, is very popular. We have retrospectively examined the incidence of delirium in 181 consecutive patients admitted to the orthogeriatric ward with hip fractures. We used the confusion assessment method to establish the presence of delirium in all patients. A database search was conducted to identify which patients were treated by Meperidine, or Morphine, prior to delirium onset. We identified 92 cases, 44 of whom were treated by Meperidine alone, and the other 48 treated by Morphine alone. Delirium was diagnosed in 13 (27.1%) Morphine treated patients as compared with 19 (43.2%) treated by Meperidine (P<0.001). Age, cognitive status and opiate use were associated with perioperative delirium. A subset regression analysis showed that exposure to Meperidine was significantly associated with delirium (odds ratio 2.5, P<0.01), in contrast with Morphine. Our results confirm the association between exposure to Meperidine and delirium, suggesting that this drug should be withdrawn in elderly hip fractured patients undergoing surgery, and substituted by low dose Morphine analgesia. Reducing the incidence of delirium, by adopting such an approach, may result in a significant potential of savings in direct costs, related to treatment of delirium in this population. PMID- 14764365 TI - Loneliness, health and social network among elderly people--a follow-up study. AB - The purpose of this study is to describe and follow up the existence of experienced loneliness, subjective health and social network among elderly people during a period of 10 years. The study is based on interviews (with structured questions) with persons, aged 75 years and over, performed by trained nurses at a baseline (in 1987) and three follow-ups. Descriptive statistics (frequency) and chi2 tests were used to study significant differences of the variables experienced loneliness, subjective health, housing, satisfaction with friend contacts and having a good friend to talk to, over time. The results showed that most elderly people tended to report high satisfaction with friend contacts over the study time, despite the decrease in having a good friend to talk to. The satisfaction with social contacts was very close connected with feelings of loneliness. Therefore, a greater understanding for the meaning of having a good friend/friends to talk to (meaningful contacts) seems to be important in caring for elderly people. PMID- 14764367 TI - Risk factors of mortality among the home-based disabled patients in central Taiwan. AB - The objective of this study was to investigate the factors that affected mortality 1 year after an intensive service program among the disabled. Three hundred and twenty-two disabled subjects were selected from communities in central Taiwan. Sixty-six percent of the participants had cerebral vascular accident (CVA) followed by hypertension (28%) and diabetes mellitus (20%). After 1 year, 117 (36.3%) had died. The activities of daily living (ADL) scores of the participants who had died were significantly lower than those in the survival group. Of those who had pressure sores, 60% were in the 'death group' compared with about 40% in the survival group. Overall, mortality rate was 36.3% after the intensive service program. Based on multiple logistic regression, the factors that affected mortality were as follows: age over 65 years compared with less than 64 years (adjust odds ratio (AOR)=1.897), severe ADL dependence (AOR=0.502), incidence of CVA (AOR=0.448), and pressure sores (AOR=1.978). The authors recommend that the factors affecting mortality be evaluated urgently and that medical services and community support systems cooperate to provide disabled patients with adequate health care. In addition, the implementation of the intensive service program provides an opportunity and challenge for nurses to examine their working practices. PMID- 14764369 TI - The meat in the diet of aged subjects and the antioxidant effects of carnosine. PMID- 14764366 TI - Urinary incontinence, its influence on daily life, and use of continence aids in two cohorts of 85/86-year-old free-living men and women, born 10 years apart. AB - The aim of this study was to assess the prevalence of urinary incontinence and other urinary tract symptoms, the use of incontinence aids and how urinary incontinence influences daily life in two 85/86-year-old cohorts born 10 years apart. There were no statistically significant differences between the two cohorts except for the use of incontinence aids, where 23% of women born in 1901/1902 and 42% of women born in 1911/1912 used aids (P=0.002). In this study, urinary incontinence was common among 85/86-year-old men and women but there were no significant differences except for the more frequent use of incontinence aids among women born in 1911/1912 than among women born in 1901/1902. The reason for this difference regarding use of incontinence aids among women could be that taboos are gradually lifting in many cultures, for example in Sweden, but so far mostly for women. PMID- 14764370 TI - The role of reality orientation therapy in restorative care of elderly patients with dementia plus stroke in the subacute nursing home setting. PMID- 14764371 TI - Institutionalized elderly, professional burden and job burnout. PMID- 14764372 TI - Evaluation of temporal and spatial disorientation in elderly patients with cognitive impairment. PMID- 14764373 TI - Analysis of hemochromatosis gene mutations in the Sicilian population: implications for survival and longevity. PMID- 14764374 TI - Electrolyte derangements and diuretic misuse in the elderly. PMID- 14764375 TI - Atrioventricular block and pacemaker implantation in a 102-year-old patient. PMID- 14764376 TI - Geriatric health care to the elderly in Israel. PMID- 14764377 TI - Prevalence of non organ-specific autoantibodies in healthy centenarians. PMID- 14764378 TI - Growth hormone (GH) and insulin-like growth factor-1 (ICF-1) in the elderly: clinical trial results. PMID- 14764379 TI - Lifestyle and cardiovascular aging in centenarians. PMID- 14764380 TI - Age- and gender-related values of lymphocyte subsets in subjects from Northern and Southern Italy. PMID- 14764381 TI - The sick euththyroidsyndrome (SES): focusing on the role of cytokines. PMID- 14764382 TI - The importance of assessment of nutritional status for the extension of an independent longevity. PMID- 14764383 TI - Plasma Abeta1-42 levels in Alzheimer's disease patients. PMID- 14764384 TI - The cardiac bradyarrhythmic syncope among the patients of a geriatric division. PMID- 14764385 TI - Correlations between the behavioral disorders in Alzheimer's disease and caregiver's distress. PMID- 14764386 TI - The aging heart and exercise training. PMID- 14764387 TI - Sildenafil in the treatment of erectile dysfunction in elderly depressed patients with idiopathic Parkinson's disease. PMID- 14764388 TI - Elderly patients with osteoarthritis: treatment with nonselective, nonsteroidial, antiinflammatory drugs, and the gastrointestinal consequences. PMID- 14764390 TI - Health perception of elderly people: the results of a longitudinal study. PMID- 14764389 TI - Advances in treatment strategies of hypertension in elderly subjects: very low dose combination in first line. PMID- 14764391 TI - Diagnostic, therapeutic, ethic and legal issues in caring for dementia: the viewpoint of medical representative in Modena (Italy). PMID- 14764392 TI - Causes of death in over-85-year-old hospitalized patients: a retrospective study. PMID- 14764393 TI - Centenarians in good health conditions. PMID- 14764394 TI - Tinnitus and oxidative stress in a selected series of elderly patients. PMID- 14764395 TI - Parkinson's disease, Parkinsonism and aging. PMID- 14764396 TI - Lack of association between ace polymorphism and Alzheimer's disease in southern Italy. PMID- 14764397 TI - Apolipoprotein and antiotensin converting enzyme genes: regional differences and extreme longevity in Europe. PMID- 14764398 TI - Aging, rheumoarthropaties and home accidents. PMID- 14764399 TI - Arthrorheumatic diseases in Sicily: Aspects of financial burden and care organization. PMID- 14764400 TI - Arthrorheumatic diseases, functional impairment and comments on the elimination of architectural barriers. PMID- 14764401 TI - Age-related changes in serum total antioxidant capacity (TAC), dehydroepiandrosterone sulphate (DHEAS), and insulin-like growth factor-1 (IGF 1): evidence in healthy centenarian subjects. PMID- 14764402 TI - Healthy centenarian subjects as living model of "successful aging": A study on fasting glycemia, C-peptide, dehydroepiandrosterone sulphate (DHEAS) and insulin like growth factor (IGF-1). PMID- 14764403 TI - Dementia due to acute hyponatremic encephalopathy in an elderly patient with Arnold-Chiari-I syndrome. PMID- 14764404 TI - Risk factors in the elderly. PMID- 14764405 TI - Magnesium and the maturation process in rats. PMID- 14764406 TI - Therapy of panic attacks in the elderly. PMID- 14764407 TI - Citalopram in the treatment of depression in the elderly. PMID- 14764408 TI - Use of amantadine in the treatment of the neurobehavioral sequelae after brain injury in elderly patients. PMID- 14764409 TI - Chronic atrial fibrillation and asymptomatic cerebral infarction in elderly patients. PMID- 14764410 TI - The use of selegiline in the treatment of cognitive deficits in elderly patients. PMID- 14764411 TI - Magnesium supplementation and age-related changes in lipid status of rats. PMID- 14764412 TI - Endothelin-1 and endothelial damage in elderly patients. PMID- 14764413 TI - Lipoprotein(a) in the elderly: beyond atherosclerosis. PMID- 14764414 TI - Serum apoliprotein E levels in alzheimer's disease and extreme longevity. PMID- 14764415 TI - Twenty-four-hour urinary cortisol levels in Alzheimer disease and in dysthymia. PMID- 14764416 TI - The elderly yesterday, today and tomorrow: aspects on cohort differences from the gerontological and geriatric population studies in Goteborg, Sweden (H70). PMID- 14764417 TI - Depression and aging: a survival study on centenarians. PMID- 14764418 TI - Validation of the Cohen-Mansfield agitation inventory (CMAI-long form) on a sample of Italian demented patients. PMID- 14764419 TI - On the reality and possibilities of pharmacological anti-aging interventions to support successful aging. PMID- 14764420 TI - Oxidized low-density lipoprotein retards the growth of proliferating cells by inhibiting nuclear translocation of cell cycle proteins. AB - OBJECTIVE: Our study tested the hypothesis that the mitogenic effect of oxidized low-density lipoprotein (oxLDL) on vascular cells may be further enhanced by the presence of cytokines and growth factors known to be present in the atherosclerotic environment. METHODS AND RESULTS: Quiescent fibroblasts and vascular smooth muscle cells were treated with 10 or 50 microg/mL minimally oxidized LDL in combination with serum for 24 or 48 hours. Surprisingly, these cells showed inhibited release from growth arrest and a significant reduction in the number of cells completing the cell cycle when compared with cells treated with serum alone. This was not due to an induction of apoptosis. The antiproliferative effects were not closely associated with changes in the expression of cell cycle proteins. Instead, oxLDL inhibited the translocation of cell cycle proteins cell division cycle (Cdc) 2, cyclin-dependent kinase (Cdk) 2, Cdk 4, Cyclin A, Cyclin B1, Cyclin D1, and proliferative cell nuclear antigen (PCNA) into the nucleus, as compared with separate treatments with serum alone. Kinase activation associated with specific cell cycle proteins was also inhibited by oxLDL. CONCLUSIONS: oxLDL, in the presence of serum, has a surprising inhibitory effect on cell proliferation that occurs through an inhibition of import of cell cycle proteins into the cell nucleus. PMID- 14764421 TI - Brain cholesterol: long secret life behind a barrier. AB - Although an immense knowledge has accumulated concerning regulation of cholesterol homeostasis in the body, this does not include the brain, where details are just emerging. Approximately 25% of the total amount of the cholesterol present in humans is localized to this organ, most of it present in myelin. Almost all brain cholesterol is a product of local synthesis, with the blood-brain barrier efficiently protecting it from exchange with lipoprotein cholesterol in the circulation. Thus, there is a highly efficient apolipoprotein dependent recycling of cholesterol in the brain, with minimal losses to the circulation. Under steady-state conditions, most of the de novo synthesis of cholesterol in the brain appears to be balanced by excretion of the cytochrome P 450-generated oxysterol 24S-hydroxycholesterol. This oxysterol is capable of escaping the recycling mechanism and traversing the blood-brain barrier. Cholesterol levels and cholesterol turnover are affected in neurodegenerating disorders, and the capacity for cholesterol transport and recycling in the brain seems to be of importance for the development of such diseases. The possibility has been discussed that administration of inhibitors of cholesterol synthesis may reduce the prevalence of Alzheimer disease. No firm conclusions can, however, be drawn from the studies presented thus far. In the present review, the most recent advances in our understanding of cholesterol turnover in the brain is discussed. PMID- 14764422 TI - Embryonic cell lines with endothelial potential: an in vitro system for studying endothelial differentiation. AB - OBJECTIVE: Endothelial differentiation is a fundamental process in angiogenesis and vasculogenesis with implications in development, normal physiology, and pathology. To better understand this process, an in vitro cellular system that recapitulates endothelial differentiation and is amenable to experimental manipulations is required. METHODS AND RESULTS: Embryonic cell lines that differentiate exclusively into endothelial cells were derived from early mouse embryos using empirical but reproducible culture techniques without viral or chemical transformation. The cells were not pluripotent and expressed reduced levels of Oct 4 and Rex-1. They were non-tumorigenic with a population doubling time of approximately 15 hours. When plated on matrigel, they readily differentiated to form patent tubular structures with diameters of 30 to 150 microm. The differentiated cells endocytosed acetylated low-density lipoprotein (LDL) and began to express endothelial-specific markers such as CD34, CD31, Flk 1, TIE2, P-selectin, Sca-1, and thy-1. They also expressed genes essential for differentiation and maintenance of endothelial lineages, eg, Flk-1, vascular endothelial growth factor (VEGF), and angiopoietin-1. When transplanted into animal models, these cells incorporated into host vasculature. CONCLUSIONS: These cell lines can undergo in vitro and in vivo endothelial differentiation that recapitulated known endothelial differentiation pathways. Therefore, they are ideal for establishing an in vitro cellular system to study endothelial differentiation. PMID- 14764423 TI - Expansive arterial remodeling: location, location, location. AB - The artery is a dynamic organ capable of changing its geometry in response to atherosclerotic plaque formation. Expansion of the vessel diameter retards luminal narrowing and is considered a compensatory response. However, the expansive remodeling response is a "wolf in sheep's clothes," because expansion is associated with the presence of inflammatory cells, proteolysis, and a thrombotic plaque phenotype. The prevalence and clinical presentation of expansively remodeled lesions may differ among vascular beds. However, it is evident that all types of atherosclerotic arterial expansive lesions share the presence of inflammatory cells and subsequent protease activities. The potential role of inflammation and protease activity in the development of the different remodeling modes is discussed. PMID- 14764425 TI - Statins after ischemic stroke and transient ischemic attack: an advisory statement from the Stroke Council, American Heart Association, and American Stroke Association. PMID- 14764424 TI - Plant sterol-fortified orange juice effectively lowers cholesterol levels in mildly hypercholesterolemic healthy individuals. AB - OBJECTIVE: Hypercholesterolemia is a major risk factor for coronary artery disease. Therapeutic lifestyle changes include dietary modifications such as inclusion of phytosterols, which effectively lowers low-density lipoprotein (LDL) cholesterol in margarines and other fats. Their effectiveness in nonfat moieties is not yet established. The aim of this study was to examine if phytosterols alter the plasma lipoprotein profile when incorporated into nonfat orange juice. METHODS AND RESULTS: After a 2-week run-in phase with orange juice, 72 mildly hypercholesterolemic healthy subjects were randomized to receive either placebo orange juice (placebo OJ) or plant sterol-fortified orange juice (sterol OJ) (2g/d) for 8 weeks. Fasting blood was obtained at baseline, after 2 weeks of OJ, and after 8 weeks of placebo/sterol-OJ supplementation. Sterol OJ supplementation significantly decreased total (7.2%), LDL (12.4%), and non-high-density lipoprotein (HDL) cholesterol (7.8%) compared with baseline and compared with placebo OJ (P<0.01). Apolipoprotein B levels were significantly decreased (9.5%) with sterol OJ. There were no significant changes in HDL cholesterol or triglycerides with the sterol OJ. While folate and B12 levels significantly increased, homocysteine levels were unchanged. CONCLUSIONS: Orange juice fortified with plant sterols are effective in reducing LDL cholesterol and could easily be incorporated into the therapeutic lifestyle changes dietary regimen. PMID- 14764426 TI - Role of p160 coactivator complex in the activation of liver X receptor. AB - OBJECTIVE: Liver X receptor (LXR) is a member of a nuclear receptor family regulating the expression of several key proteins involved in lipid metabolism and inflammation. In contrast to several other nuclear receptors, very little is known about the coactivators needed for the agonist-mediated transactivation by LXR. In this study, we have investigated the role of p160 coactivator complex in the regulation of ATP-binding transporter A1 (ABCA1), a clinically important gene transcriptionally upregulated by LXR/RXR (retinoid X receptor) heterodimer. METHODS AND RESULTS: Overexpression of LXRalpha, SRC-1, and p300, either alone or in combination, increased the luciferase activity driven by the wild-type ABCA1 promoter. The same coactivators bound to the ABCA1 promoter on oxysterol induction in chromatin immunoprecipitation assays. To the contrary, CARM-1 and P/CAF had no effect on ABCA1 transactivation, nor do they bind the promoter. When the DR-4 element was mutated from the ABCA1 promoter, only p300 was able to activate ABCA1 transcription in a ligand-independent manner. CONCLUSIONS: The p160 coactivator complex members SRC-1 and p300, but not CARM-1 and P/CAF, coactivate LXR-mediated transcription of ABCA1 gene. In addition, p300 activates ABCA1 transcription independently of DR-4 element and LXR/RXR. PMID- 14764427 TI - Regulation of urokinase receptor expression by phosphoglycerate kinase. AB - Post-transcriptional regulation represents a major mechanism by which eukaryotic gene expression is regulated through cis-trans interactions that serve as signals for rapid alterations of messenger RNA (mRNA) stability. Regulation of urokinase type plasminogen activator receptor (uPAR) mRNA involves the interaction of a uPAR mRNA coding region sequence with a 50 kD uPAR mRNA binding protein. We purified this protein from human bronchial epithelial (Beas2B) cells and identified it as phosphoglycerate kinase (PGK). We cloned PGK cDNA by polymerase chain reaction and expressed the recombinant PGK protein, which specifically bound the uPAR mRNA coding region by gel mobility shift and Northwestern blotting. We also confirmed a direct interaction of PGK protein with uPAR mRNA by immunoprecipitation. Overexpression of PGK in uPAR-overproducing H157 lung carcinoma cells resulted in decreased cytoplasmic uPAR mRNA and cell surface uPAR protein expression. Reduced uPAR mRNA expression involved decreased stability of the uPAR mRNA. Decline in 3H-thymidine incorporation and migration occurred in H157 cells transfected with PGK cDNA. These results demonstrate that PGK regulates uPAR expression at the post-transcriptional level. PMID- 14764428 TI - Modulation of calcium signaling by interleukin-13 in human airway smooth muscle: role of CD38/cyclic adenosine diphosphate ribose pathway. AB - CD38/cyclic adenosine diphosphate ribose (cADPR) signaling plays an important role in the regulation of intracellular calcium responses to agonists in a variety of cells, including airway smooth muscle (ASM) cells. The present study was aimed at determining the effect of interleukin (IL)-13, a cytokine implicated in the pathogenesis of asthma, on CD38/cADPR signaling and to ascertain the contribution of CD38/cADPR signaling to IL-13-induced airway hyperresponsiveness. Human ASM cells maintained in culture were exposed to 50 ng/ml IL-13 for 22 h and levels of CD38 expression and intracellular calcium responses to agonists were measured. Treatment of human ASM cells with IL-13 resulted in increased CD38 expression as determined by real-time polymerase chain reaction, Western blot analysis, and indirect immunofluorescence. Increased CD38 expression was reflected as increased ADP-ribosyl cyclase activity in the ASM cell membranes. The net intracellular calcium responses to bradykinin, thrombin, and histamine were significantly (P < or = 0.05) higher in cells treated with IL-13 compared with controls. Furthermore, 8-bromo-cADPR, a cADPR antagonist, attenuated IL-13 induced augmented intracellular calcium responses to agonists in human ASM cells. These findings indicate that the CD38/cADPR-dependent pathway has a major role in IL-13-induced modulation of calcium signaling in human ASM. PMID- 14764430 TI - Pressure-volume curve does not predict steady-state lung volume in canine lavage lung injury. AB - To better understand strategies for recruiting and maintaining lung volume in acute lung injury, we examined relationships between steady-state lung volume and cumulative cyclic recruitment/derecruitment volume history and the quasi-static pressure-volume curve, in an animal saline lavage lung injury model. Small-volume tidal pressure-volume loops performed after inflation from functional residual capacity demonstrated incremental, cyclic recruitment only if the peak pressure achieved exceeded the pressure at which the compliance increased (Pflex) on the pressure-volume curve, whereas loops performed after deflation from total lung capacity remained close to the envelope deflation curve. Recruitment continued to occur up to and beyond a peak inspiratory airway pressure of 40 cm H(2)O, as demonstrated by both the tidal loops and by computed tomography-derived lung volume data. Tidal-specific compliance was relatively constant across positive end-expiratory pressure levels after inflation from functional residual capacity, but peaked at moderate positive end-expiratory pressure after deflation from total lung capacity, further demonstrating the effects of volume history and providing experimental validation of the recruitment models of Hickling (AJRCCM 2001;163:69-78). These results support the interpretation of Pflex as pressure threshold for recruitment, but otherwise do not suggest a role for the pressure volume curve in predicting steady-state lung volume. PMID- 14764429 TI - Pro- and anti-inflammatory factors cooperate to control hyaluronan synthesis in lung fibroblasts. AB - Hyaluronan (HA) is an important constituent of the extracellular matrix and accumulates during inflammatory lung diseases like asthma. Little is known about the factors that regulate HA synthesis by lung cells. Accordingly, we investigated the effect of T-helper 1 (TH1) and 2 (TH2) cytokines and the anti inflammatory agents fluticasone and salmeterol on HA synthesis in human lung fibroblasts. Interleukin-1beta (IL-1beta) and tumor necrosis factor (TNF)-alpha were the most potent stimulators of HA synthesis and when combined, caused synergistic increases in HA accumulation. Time-course analysis of HA accumulation and [3H]-glucosamine incorporation into HA demonstrated continued synthesis over the 24 h of stimulation. Peak synthesis at 6-12 h coincided with an increased proportion of high molecular weight HA. Reverse transcriptase polymerase chain reaction (RT-PCR) revealed that IL-1beta and TNF-alpha induced HA synthase-2 messenger RNA (mRNA) 3 h following stimulation and remained elevated throughout the 24-h stimulation period. Fluticasone inhibited IL-1beta and TNF-alpha induced HA synthesis (44.5%) whereas salmeterol had no effect. When combined, fluticasone and salmeterol inhibited HA synthesis to a greater extent (85.2%). Further, fluticasone attenuated IL-1beta and TNF-alpha stimulated hyaluronan synthase-2 messenger RNA (mRNA), and the addition of salmeterol cooperatively enhanced this inhibition. These results indicate that enhanced synthesis of HA by the proinflammatory cytokines IL-1beta and TNF-alpha can be abrogated by specific corticosteroid and beta2 blocker combinations shown to be effective in the treatment of asthma. PMID- 14764431 TI - The relationship between infant airway function, childhood airway responsiveness, and asthma. AB - The relationship between reduced pulmonary function in early life and persistent wheeze (PW) in school-aged children remains uncertain. In this study, VmaxFRC was assessed at 1 month of age, and the presence of wheeze up to 11 years of age was prospectively identified. At 11 years of age, airway responsiveness (AR) to inhaled histamine and atopy were assessed. Recent wheeze at 11 years of age was associated with a reduced mean z score for VmaxFRC at 1 month of age (-0.41 [SD 0.91], n = 31) compared with no recent wheeze (0.04 [SD 1.00], n = 153, p = 0.03). Wheeze between 4 and 6 years that persisted at 11 years (PW) was most prevalent among those with reduced VmaxFRC at 1 month and atopy aged 11 years (p = 0.002) or reduced VmaxFRC and increased AR aged 11 years (p = 0.015). When all factors were considered, reduced VmaxFRC at 1 month (p = 0.03) and increased AR aged 11 years (p < 0.001) were independently associated with PW (n = 17) compared with other outcomes (n = 129). Reduced airway function present in early infancy is associated with PW at 11 years of age, and this relationship is independent of the effect of increased AR and atopy in childhood. PMID- 14764432 TI - Regional effects of selective pharyngeal muscle activation on airway shape. AB - Pharyngeal airway fiberoptic imaging was performed in 10 decerebrate cats to determine the effect of selective pharyngeal muscle activation on airway shape. At intraluminal pressures from 6 to -6 cm H2O, maximum anteroposterior and lateral diameters were measured in the rostral oropharynx, caudal oropharynx, and velopharynx with and without bilateral stimulation of the medial hypoglossus (HG), lateral HG, whole HG, glossopharyngeus, and pharyngeal branch of vagus nerves. At all three airway levels without nerve stimulation, the increase in diameter with increasing pressure was greater in the lateral than anteroposterior dimension. Stimulation of the hypoglossal and glossopharyngeus nerves caused greater increases in lateral than anteroposterior diameter in all three regions with different effects across nerves and regions. Stimulation of these four nerves frequently caused greater increases in both diameters, as the airway cross sectional area was decreased by lowering airway pressure. Stimulation of the pharyngeal branch of vagus resulted in greater decreases in lateral than anteroposterior dimension in the caudal oropharynx and velopharynx, especially as airway cross-sectional area was increased by increasing intraluminal pressure. The results indicate that selective activation of pharyngeal muscles in cats frequently results in greater changes in lateral than anteroposterior airway diameter and that these effects are dependent on airway region and cross sectional area. PMID- 14764433 TI - Twenty-four-hour ambulatory blood pressure in children with sleep-disordered breathing. AB - Obstructive sleep apnea causes intermittent elevation of systemic blood pressure (BP) during sleep. To determine whether obstructive apnea in children has a tonic effect on diurnal BP, 24-hour ambulatory blood pressure was obtained from 60 children with mean age of 10.8 +/- 3.5 years. Thirty-nine children had obstructive apnea and 21 had primary snoring. Children with obstructive apnea had significantly greater mean BP variability during wakefulness and sleep, a higher night-to-day systolic BP, and a smaller nocturnal dipping of mean BP. Variability of mean arterial pressure during wakefulness was predicted by the desaturation, body mass, and arousal indices, whereas variability during sleep was predicted by apnea-hypopnea and body mass indices. Nocturnal BP dipping was predicted by the desaturation index. There were no significant differences in systolic, diastolic, or mean arterial BP during sleep between the groups. Diastolic BP during wakefulness was significantly different between the groups and correlated negatively with apnea-hypopnea index. We conclude that obstructive apnea in children is associated with 24-hour BP dysregulation and that, independent of obesity, the frequency of obstructive apnea, oxygen desaturation, and arousal contributes to abnormal BP control. PMID- 14764434 TI - Chemical activation of C1-C2 spinal neurons modulates intercostal and phrenic nerve activity in rats. AB - Chemical activation of upper cervical spinal neurons modulates activity of thoracic respiratory interneurons in rats. The aim of the present study was to examine the effects of chemical activation of C(1)-C(2) spinal neurons on thoracic spinal respiratory motor outflows. Electroneurograms of left phrenic (n = 23) and intercostal nerves (ICNs, n = 93) between T(3) and T(8) spinal segments were recorded from 36 decerebrated, vagotomized, paralyzed, and ventilated male rats. To activate upper cervical spinal neurons, glutamate pledgets (1 M, 1 min) were placed on the dorsal surface of the C(1)-C(2) spinal cord. Glutamate on C(1) C(2) increased ICN tonic activity in 56/59 (95%) ICNs. The average maximal tonic activity of ICN was increased by 174% (n = 59). After spinal transection at rostral C(1), glutamate on C(1)-C(2) still increased ICN tonic activity in 33/35 ICNs. However, the effects of C(1)-C(2) glutamate on ICN phasic activity were highly variable, with observations of augmentation or suppression of both inspiratory and expiratory discharge. C(1)-C(2) glutamate augmented the average amplitude of phrenic burst by 20%, whereas the increases in amplitude of ICN inspiratory activity, when they occurred, averaged 120%. The burst rate of phrenic nerve discharge was decreased from 34.2 +/- 1.6 to 26.3 +/- 2.0 (mean +/- SE) breaths/min during C(1)-C(2) glutamate. These data suggested that upper cervical propriospinal neurons might play a role in descending modulation of thoracic respiratory and nonrespiratory motor activity. PMID- 14764435 TI - Gestational and early postnatal dietary NaCl levels affect NaCl intake, but not stimulated water intake, by adult rats. AB - We examined body fluid regulation by weanling (21-25 days) and adult (>60 days) male rats that were offspring of dams fed chow containing either 0.1, 1, or 3% NaCl throughout gestation and lactation. Weanling rats were maintained on the test diets until postnatal day 30 and on standard 1% NaCl chow thereafter. Ad libitum water intake by weanlings was highest in those fed 3% NaCl and lowest in those fed 0.1% NaCl. Adult rats maintained on standard NaCl chow consumed similar amounts of water after overnight water deprivation or intravenous hypertonic NaCl (HS) infusion regardless of early NaCl condition. Moreover, baseline and HS stimulated plasma Na(+) concentrations also were similar for the three groups. Nonetheless, adult rats in the early 3% NaCl group consumed more of 0.5 M NaCl after 10 days of dietary Na(+) deprivation than did rats in either the 1% or 0.1% NaCl group. Interestingly, whether NaCl was consumed in a concentrated solution in short-term, two-bottle tests after dietary Na(+) deprivation or in chow during ad libitum feeding, adult rats in the 3% NaCl group drank less water for each unit of NaCl consumed, whereas rats in the 0.1% NaCl group drank more water for each unit of NaCl consumed. Thus gestational and early postnatal dietary NaCl levels do not affect stimulated water intake or long-term body fluid regulation. Together with our previous studies, these results suggest that persistent changes in NaCl intake and in water intake associated with NaCl ingestion reflect short term behavioral effects that may be attributable to differences in NaCl taste processing. PMID- 14764436 TI - Effects of dietary calcium restriction and acute exercise on the antioxidant enzyme system and oxidative stress in rat diaphragm. AB - Calcium deficiency is considered to increase intracellular calcium level; thus the aim of the current study was to elucidate whether dietary calcium restriction enhanced exercise-induced oxidative stress in rat diaphragm. Twenty male Wistar rats were randomly assigned to either a control group or a group subjected to 1 mo of calcium restriction. In addition, each group was subsequently subdivided into rested or acutely exercised group. Dietary calcium restriction significantly (P < 0.05) upregulated the activities of manganese-superoxide dismutase (Mn-SOD), copper-zinc-superoxide dismutase (Cu-Zn-SOD), and glutathione peroxidase (Gpx) but not catalase. Acute exercise, in addition to calcium restriction, decreased both SOD isoenzymes in the diaphragm of calcium-restricted rats (P < 0.05). On the other hand, calcium restriction resulted in increased Gpx mRNA expression (P < 0.05). In control rats, acute exercise significantly (P < 0.05) increased the expressions of both SOD mRNAs, whereas in the calcium-restricted rats, it increased that of Mn-SOD mRNA (P < 0.05) but decreased that of Gpx mRNA (P < 0.05). Furthermore, reactive carbonyl derivative, a marker of protein oxidation, was significantly greater in the calcium-restricted rats than in the control rats after acute exercise (P < 0.05). The results suggest that antioxidant enzymes in rat diaphragm were upregulated in response to an increased oxidative stress by dietary calcium restriction but that upregulation is not enough to cope with exercise-induced further increase of oxidative stress. PMID- 14764437 TI - Acute hemodynamic responses in the head during microgravity induced by free drop in anesthetized rats. AB - To examine acute hemodynamic responses to microgravity (microG) in the head, we measured carotid artery pressure (CAP) and jugular vein pressure (JVP) to calculate cephalic perfusion pressure (CPP = CAP - JVP) and recorded images of microvessels in the iris to evaluate capillary blood flow velocity (CBFV) and capillary diameter (CD) in anesthetized rats during 4.5 s of microG induced by free drop. Rats were placed in 30 degrees head-up whole body-tilted (HU, n = 7) or horizontal (flat, n = 6) position. In the flat group, none of the measured variables was significantly affected by microG, whereas in the HU group, CAP, JVP, and CPP increased, respectively, by 23.4 +/- 2.6, 1.3 +/- 0.2, and 22.9 +/- 3.1 mmHg, and CBFV and CD increased, respectively, by 33 +/- 8 and 9 +/- 3%, showing an increase in capillary blood flow. To further examine the mechanisms underlying these CAP and JVP increases, another experiment was performed in which CAP and JVP were measured in anesthetized rats (n = 6) during a postural change from HU to flat. In these animals, the change in JVP was similar to that observed during actual microG, but no change in CAP was seen, indicating that the JVP increase during actual microG is caused by disappearance of the gravitational pressure gradient in the head-to-foot axis, whereas the CAP increase is not. In conclusion, actual microG elicits an increase in CPP due to a greater increase in CAP than JVP, resulting in increased capillary blood flow. Although the increase in JVP is explained by the disappearance of gravitational pressure gradient in the head-to-foot axis as a result of microG, the larger increase in CAP is not. PMID- 14764438 TI - A novel framework of circulatory equilibrium. AB - A novel framework of circulatory equilibrium was developed by extending Guyton's original concept. In this framework, venous return (CO(V)) for a given stressed volume (V) was characterized by a flat surface as a function of right atrial pressure (P(RA)) and left atrial pressure (P(LA)) as follows: CO(V) = V/W - G(S)P(RA) - G(P)P(LA), where W, G(S), and G(P) denote linear parameters. In seven dogs under total heart bypass, CO(V), P(RA), P(LA), and V were varied to determine the three parameters in each animal with use of multivariate analysis. The coefficient of determination (r(2) = 0.92-0.99) indicated the flatness of the venous return surface. The averaged surface was CO(V) = V/0.129 - 19.61P(RA) - 3.49P(LA). To examine the invariability of the surface parameters among animals, we predicted the circulatory equilibrium in response to changes in stressed volume in another 12 dogs under normal and heart failure conditions. This was achieved by equating the standard surface with the individually measured cardiac output (CO) curve. In this way, we could predict CO [y = 0.90x + 5.6, r(2) = 0.95, standard error of the estimate (SEE) = 8.7 ml.min(-1).kg(-1)], P(RA) (y = 0.96x, r(2) = 0.98, SEE = 0.2 mmHg), and P(LA) (y = 0.89x + 0.5, r(2) = 0.98, SEE = 0.8 mmHg) reasonably well. We conclude that the venous return surface accurately represents the venous return properties of the systemic and pulmonary circulations. The characteristics of the venous return surface are invariable enough among animals, making it possible to predict circulatory equilibrium, even if those characteristics are unknown in individual animals. PMID- 14764439 TI - Transcription activator protein 1 mediates alpha- but not beta-adrenergic hypertrophic growth responses in adult cardiomyocytes. AB - In some models of cardiac hypertrophy, activation of activator protein 1 (AP-1) correlates with growth. However, AP-1 is also activated by stimuli not involved in cardiac growth. This raises the following questions: does AP-1 plays a causal role for cardiomyocyte growth, and is this role model or stimulus dependent? We used a single model to address these questions, i.e., ventricular cardiomyocytes of adult rats, and two growth stimuli, i.e., alpha- and beta-adrenoceptor agonists [10 microM phenylephrine (PE) and 1 microM isoprenaline (Iso), respectively]. After 1 h of stimulation with PE, mRNA expression of c-Fos and c Jun was upregulated to 185 +/- 32 and 132 +/- 13% of control. Fos and Jun proteins formed the AP-1 complex. PE stimulated DNA binding activity of AP-1 to 165 +/- 22% of control within 2 h and increased protein synthesis to 161 +/- 27% of control and cross-sectional area to 126 +/- 4% of control. Inhibition of AP-1 binding activity by cAMP response element (CRE) decoy oligonucleotides abolished both of these growth responses. Iso stimulated AP-1 binding activity to 203 +/- 19% of control within 2 h and stimulated protein synthesis to 145 +/- 17% of control. However, the growth effect of Iso was not abolished by CRE decoys: Iso increased protein synthesis to 158 +/- 17% of control in the presence of CRE. In conclusion, AP-1 is a causal mediator of the alpha-adrenergic, but not the beta adrenergic, growth response of cardiomyocytes. PMID- 14764440 TI - Presynaptic modulation of evoked NE release contributes to sympathetic activation after pressure overload. AB - Activation of the sympathetic nervous system is well documented in heart failure. Our previous studies demonstrated an increase in evoked norepinephrine (NE) release from left ventricle (LV) slices at 10 days of pressure overload. The purpose of this study was to test the hypothesis that presynaptic modulation of NE release contributes to sympathetic activation after pressure overload. We examined the functional status of the presynaptic alpha(2)- and beta(2)-receptors and ANG II subtype 1 (AT(1)) receptors in LV slices from 10-day aortic constricted (AC) and sham-operated (SO) rats. Evoked (3)H overflow from LV slices preloaded with [(3)H]NE was increased in AC rats. The alpha(2)-agonist UK-14,304 decreased evoked (3)H overflow with no differences between groups. The beta(2) agonist salbutamol increased evoked (3)H overflow with greater sensitivity in slices from AC rats. The beta-antagonist propranolol decreased evoked (3)H overflow from LV slices of AC rats but not controls. ANG II increased evoked (3)H overflow with greater sensitivity in slices from AC rats. These data support the hypothesis that aberrant presynaptic modulation of catecholamine release contributes to sympathetic activation after pressure overload. PMID- 14764441 TI - Direct effect of ethanol on human vascular function. AB - Epidemiological studies indicate that moderate ethanol consumption reduces cardiovascular mortality. Cellular and animal data suggest that ethanol confers beneficial effects on the vascular endothelium and increases the bioavailability of nitric oxide. The purpose of this study was to assess the effect of ethanol on endothelium-dependent, nitric oxide-mediated vasodilation in healthy human subjects. Forearm blood flow (FBF) was determined by venous occlusion plethysmography in healthy human subjects during intra-arterial infusions of either methacholine (0.3, 1.0, 3.0, and 10.0 mcg/min, n = 9), nitroprusside (0.3, 1.0, 3.0, and 10.0 mcg/min, n = 9), or verapamil (10, 30, 100, and 300 mcg/min, n = 8) before and during the concomitant intra-arterial infusions of ethanol (10% ethanol in 5% dextrose). Additionally, a time control experiment was conducted, during which the methacholine dose-response curve was measured twice during vehicle infusions (n = 5). During ethanol infusion, mean forearm and systemic alcohol levels were 227 +/- 30 and 6 +/- 0 mg/dl, respectively. Ethanol infusion alone reduced FBF (2.5 +/- 0.1 to 1.9 +/- 0.1 ml.dl(-1).min(-1), P < 0.05). Despite initial vasoconstriction, ethanol augmented the FBF dose-response curves to methacholine, nitroprusside, and verapamil (P < 0.01 by ANOVA for each). To determine whether this augmented FBF response was related to shear-stress-induced release of nitric oxide, FBF was measured during the coinfusion of ethanol and N(G)-nitro-L-arginine (L-NAME; n = 8) at rest and during verapamil-induced vasodilation. The addition of L-NAME did not block the ability of ethanol to augment verapamil-induced vasodilation. Ethanol has complex direct vascular effects, which include basal vasoconstriction as well as potentiation of both endothelium-dependent and -independent vasodilation. None of these effects appear to be mediated by an increase in nitric oxide bioavailability, thus disputing findings from preclinical models. PMID- 14764442 TI - Gastrin regulates the heparin-binding epidermal-like growth factor promoter via a PKC/EGFR-dependent mechanism. AB - Gastrin is a known growth/differentiation factor for the gastric mucosa. Its effects are likely mediated by the induction of heparin-binding epidermal-like growth factor (HB-EGF), a member of the EGF family of growth factors that is expressed by gastric parietal cells. In this study, we investigated the regulation of the HB-EGF promoter by gastrin in a human gastric cancer cell line. Serial human HB-EGF promoter-luciferase reporter deletion constructs and heterologous promoter constructs were transfected into AGS-E cells and stimulated with gastrin (10(-7) M) with or without various signal transduction inhibitors. EMSA were also performed. Gastrin stimulation resulted in a fivefold increase in HB-EGF-luciferase activity. The cis-acting element mediating gastrin responsiveness was mapped to the -69 to -58 region of the HB-EGF promoter. Gastrin stimulation was PKC dependent and at least partially mediated by activation of the EGF receptor. PMID- 14764444 TI - Cholecystokinin pathways modulate sensations induced by gastric distension in humans. AB - Ingested fat releases CCK, causes gastric relaxation, delays gastric emptying, and limits meal size; however, the mechanistic link among these actions has not been established. Fatty acid release of CCK is chain-length sensitive; dodecanoic acid (C12) induces greater CCK release than decanoic acid (C10). The effect of C12 or C10 on tolerance to subsequent intragastric infusion of liquid was determined in healthy subjects, with and without the CCK(1) receptor antagonist dexloxiglumide. Gastric wall relaxation after either fatty acid was assessed by graded volume distension and by barostat; gastric emptying was measured by gastric aspiration and by a [(13)C]octanoic acid breath technique. C12 released more CCK (mean plasma CCK after vehicle, 4.7 +/- 0.8 pM; C10, 4.8 +/- 0.3 pM; C12, 8 +/- 1.2 pM; P < 0.05 C12 vs. C10 or vehicle) and reduced the volume of water (and of 5 and 25% glucose solutions) delivered at maximum tolerance compared with C10 or vehicle (volume of water tolerated after vehicle, 1,535 +/- 164 ml; C10, 1,335 +/- 160 ml; C12, 842 +/- 103 ml; P < 0.05 C12 vs. C10 or vehicle); this effect was abolished by dexloxiglumide. Intragastric volumes were always similar at the limit of tolerance, and, whereas gastric relaxation occurred to similar degrees after the fatty acids, its duration was longer after C12, which also induced a longer delay in half-gastric emptying [t(1/2)(min) after vehicle, 53 +/- 2; C10, 67 +/- 3; C12, 88 +/- 7; P < 0.05 C12 vs. C10 or vehicle]. In conclusion, ingestion of a CCK-releasing fatty acid reduces the tolerated volume of liquid delivered into the stomach, primarily via a CCK(1) receptor-mediated delay in gastric emptying. PMID- 14764443 TI - Expression of P2Y nucleotide receptors and ectonucleotidases in quiescent and activated rat hepatic stellate cells. AB - Extracellular nucleotides regulate a variety of cellular activities, including proliferation of fibrogenic cells outside of the liver. However, the expression of receptors for extracellular nucleotides in hepatic stellate cells (HSC) is unknown. Thus our aims were to investigate the expression of mediators of nucleotide signaling in HSC and to determine whether extracellular nucleotides regulate HSC function. Confocal video microscopy was used to observe nucleotide induced changes in cytosolic Ca(2+) (Ca(i)(2+)) in live HSC. P2Y receptor subtype expression and ectonucleotidase expression in quiescent and activated HSC were determined using RT-PCR, Northern blot, immunoblot, and confocal immunofluorescence. Functional ectonucleotidase activity was assessed using a colorimetric method. Nucleotide-sensitive procollagen-1 mRNA expression in activated HSC was assessed using real-time RT-PCR. Extracellular ATP increased Ca(i)(2+) in HSC; this was inhibited by the P2 receptor inhibitor suramin. Quiescent HSC expressed the P2Y subtypes P2Y(2) and P2Y(4) and were activated by ATP and UTP, whereas activated HSC expressed the P2Y subtype P2Y(6) and were activated by UDP and ATP. Activated but not quiescent HSC expressed the ectonucleotidase nucleoside triphosphate diphosphohydrolase 2, extracellular UDP tripled procollagen-1 mRNA expression in activated HSC, and this was inhibited by the P2Y receptor inhibitor suramin. HSC express functional P2Y receptors and switch the expression of P2Y receptor subtypes on activation. Moreover, HSC differentially regulate nucleoside triphosphate diphosphohydrolase expression after activation. Because activation of P2Y receptors in activated HSC regulates procollagen-1 transcription, P2Y receptors may be an attractive target to prevent or treat liver fibrosis. PMID- 14764445 TI - Mucosal angiogenesis regulation by CXCR4 and its ligand CXCL12 expressed by human intestinal microvascular endothelial cells. AB - Mice genetically deficient in the chemokine receptor CXCR4 or its ligand stromal cell-derived factor (SDF)-1/CXCL12 die perinatally with marked defects in vascularization of the gastrointestinal tract. The aim of this study was to define the expression and angiogenic functions of microvascular CXCR4 and SDF 1/CXCL12 in the human intestinal tract. Studies of human colonic mucosa in vivo and primary cultures of human intestinal microvascular endothelial cells (HIMEC) in vitro showed that the intestinal microvasculature expresses CXCR4 and its cognate ligand SDF-1/CXCL12. Moreover, SDF-1/CXCL12 stimulation of HIMEC triggers CXCR4-linked G proteins, phosphorylates ERK1/2, and activates proliferative and chemotactic responses. Pharmacological studies indicate SDF-1/CXCL12 evokes HIMEC chemotaxis via activation of ERK1/2 and phosphoinositide 3-kinase signaling pathways. Consistent with chemotaxis and proliferation, endothelial tube formation was inhibited by neutralizing CXCR4 or SDF-1/CXCL12 antibodies, as well as the ERK1/2 inhibitor PD-98059. Taken together, these data demonstrate an important mechanistic role for CXCR4 and SDF-1/CXCL12 in regulating angiogenesis within the human intestinal mucosa. PMID- 14764447 TI - Low-dose PGE2 mimics the duodenal secretory response to luminal acid in mice. AB - Luminal exposure to concentrated acid, the most accepted physiological stimulus for duodenal bicarbonate secretion (DBS), cannot be used with in vitro preparations due to potential tissue damage. We thus examined whether exposure to PGE(2), a well-characterized physiological duodenal secretagogue, could mimic the effects of acid perfusion. DBS was measured in C57/BL mice by pH-stat/back titration and measurement of total dissolved CO(2) concentration ([CO(2)](t)). Anion transport inhibitor DIDS, anion channel inhibitor 5-nitro-2-(3 phenylpropylamino)benzoic acid (NPPB), carbonic anhydrase inhibitor methazolamide, and nonselective cyclooxygenase inhibitor indomethacin were used to inhibit separate components of HCO(3)(-) secretory pathway. Baseline DBS was not altered by exposure to methazolamide (0.1 mM) but was slightly reduced by DIDS (0.5 mM). DBS and [CO(2)](t) increased after acid and PGE(2) exposure. DIDS (0.5 mM) and NPPB (0.2 mM) abolished acid-induced DBS increase. Methazolamide (0.1 mM) and DIDS inhibited acid-induced [CO(2)](t) increase. DIDS, NPPB, or methazolamide had little effect on DBS in response to high concentration PGE(2) (100 microg/ml). Low concentration PGE(2) (1 microg/ml) increased DBS that was inhibited by DIDS, NPPB, and methazolamide. Pretreatment with indomethacin (5 mg/kg) inhibited DBS induced by acid exposure but not by PGE(2). High-dose PGE(2) substantially increases DBS by a mechanism that appears to be different than secretory response to luminal acid perfusion. Secretory response to low-dose PGE(2), at least in terms of inhibitor profile, closely resembles secretion in response to perfusion of physiological acid concentrations and may be a useful stimulus for in vitro study of DBS in isolated mouse duodenum. PMID- 14764448 TI - Lateral intercellular space volume as a determinant of CFTR-mediated anion secretion across small intestinal mucosa. AB - Studies of full-thickness, small intestinal preparations have shown that maximal anion secretion [indexed by short-circuit current (I(sc))] during intracellular cAMP (cAMP(i)) stimulation is transient and followed by a decline toward baseline. Declining I(sc) is preceded by decreases in transepithelial conductance (G(t)), which in the small intestine reflects the lateral intercellular space (LIS) volume of the paracellular pathway. We hypothesized that decreases in LIS volume limit the magnitude and duration of cAMP(i)-stimulated anion secretion. Experimental manipulations to increase the patency of the LIS (assessed by G(t) and electron microscopy) were investigated for an effect on the magnitude of cAMP(i)-stimulated anion secretion (assessed by the I(sc) and isotopic fluxes) across murine small intestine. In control studies, changes of G(t) after cAMP(i) stimulation were associated with a morphological "collapse" of the LIS, which did not occur in intestine of CFTR-null mice. Removal of the outer intestinal musculature, exposure to a serosal hypertonic solution, or increased serosal hydrostatic pressure minimized reductions in G(t) and increased the cAMP(i) stimulated I(sc) response. Increased I(sc) primarily resulted from increased Cl( ) secretion that was largely bumetanide sensitive. However, bumetanide insensitive I(sc) was also increased, and similar increases occurred in the Na(+) K(+)-2Cl(-) cotransporter (NKCC1)-null intestine, indicating that activities of non-NKCC1 anion uptake proteins are also affected by LIS volume. Thus LIS patency is an important determinant of the magnitude and duration of CFTR-mediated anion secretion in murine small intestine. Decreases in LIS volume may limit the pool of available anions to basolateral transporters involved in transepithelial secretion. PMID- 14764449 TI - Mitogen-activated protein kinases regulate COX-2 and mucosal recovery in ischemic injured porcine ileum. AB - Mitogen-activated protein kinase (MAPK) pathways transduce signals from a diverse array of extracellular stimuli. The three primary MAPK-signaling pathways are the extracellular regulated kinases (ERK1/2), p38 MAPK, and c-Jun NH(2)-terminal kinase (JNK). Previous research in our laboratory has shown that COX-2-elaborated prostanoids participate in recovery of mucosal barrier function in ischemic injured porcine ileum. Because COX-2 expression is regulated in part by MAPKs, we postulated that MAPK pathways would play an integral role in recovery of injured mucosa. Porcine mucosa was subjected to 45 min of ischemia, after which tissues were mounted in Ussing chambers, and transepithelial electrical resistance (TER) was monitored as an index of recovery of barrier function. Treatment of tissues with the p38 MAPK inhibitor SB-203580 (0.1 mM) or the ERK1/2 inhibitor PD-98059 (0.1 mM) abolished recovery. Western blot analysis revealed that SB-203580 inhibited upregulation of COX-2 that was observed in untreated ischemic-injured mucosa, whereas PD-98059 had no effect on COX-2 expression. Inhibition of TER recovery by SB-203580 or PD-98059 was overcome by administration of exogenous prostaglandin E(2) (1 microM). The JNK inhibitor SP-600125 (0.1 mM) significantly increased TER and resulted in COX-2 upregulation. COX-2 expression appears to be positively and negatively regulated by the p38 MAPK and the JNK pathways, respectively. Alternatively, ERK1/2 appear to be involved in COX-2-independent reparative events that remain to be defined. PMID- 14764450 TI - Bacterial activation of beta-catenin signaling in human epithelia. AB - The mucosal lining of the human intestine is constantly bathed in a milieu of commensal gut flora, the vast majority of these being nonpathogenic microorganisms. Here, we demonstrate that microbial-epithelial cell interactions not only affect proinflammatory pathways but also influence beta-catenin signaling, a key component in regulating epithelial cell proliferation. The nonpathogenic Salmonella strain PhoP(c) activates the beta-catenin signaling pathway of human epithelia via a blockade of beta-catenin degradation. Normal beta-catenin ubiquitination necessary for constitutive beta-catenin degradation is abolished, allowing the accumulation and translocation of beta-catenin to the nucleus. Transcriptional activation mediated by the beta-catenin/T cell factor complex increases c-myc expression and enhances cell proliferation. We also show that the Salmonella effector protein AvrA is involved in modulating this beta catenin activation. These data suggest that nonvirulent bacterial-epithelial interactions can influence beta-catenin signaling and cell growth control in a manner previously unsuspected. PMID- 14764451 TI - Estimated global transmural distribution of activation rate and conduction block during porcine and canine ventricular fibrillation. AB - We quantified ventricular fibrillation (VF) activation rate, conduction block, and organization transmurally in pigs and dogs, whose transmural Purkinje distribution differ. In six pigs and five dogs, 75 to 100 plunge needles, containing four electrodes for the right ventricle (RV) and six electrodes for the left ventricle (LV) and septum, were inserted in vivo. Six VF episodes were electrically initiated and allowed to last for 47 to 180 seconds. From the FFT power spectra, dominant frequency (DF), an estimate of activation rate, and incidence of double peaks (DPI), an estimate of conduction block, were calculated every 8 ms at each electrode. DF was highest at the epicardium and lowest at the endocardium, whereas DPI was highest at the endocardium and lowest at the epicardium for the entire LV and the RV base in both pigs and dogs for the first 70 seconds of VF. This distribution changed little throughout the first 3 minutes of VF in pigs but reversed in dogs by 2 minutes of VF. In conclusion, estimated activation rates and conduction block incidence during VF are not uniformly distributed transmurally. During the first minute of VF, the faster activating LV base epicardium exhibits less estimated block than the slower endocardium, raising the possibility that faster activating epicardium generates wavefronts that drive the endocardium early during VF. Constancy of this pattern in pigs but its reversal by 2 minutes in dogs is consistent with the hypothesis that activation during later VF is driven by Purkinje fibers. PMID- 14764452 TI - Homeobox A9 transcriptionally regulates the EphB4 receptor to modulate endothelial cell migration and tube formation. AB - Homeobox genes (Hox) encode for transcription factors, which regulate cell proliferation and migration and play an important role in the development of the cardiovascular system during embryogenesis. In this study, we investigated the role of HoxA9 for endothelial cell migration and angiogenesis in vitro and identified a novel target gene, the EphB4 receptor. Inhibition of HoxA9 expression decreased endothelial cell tube formation and inhibited endothelial cell migration, suggesting that HoxA9 regulates angiogenesis. Because Eph receptor tyrosine kinases importantly contribute to angiogenesis, we examined whether HoxA9 may transcriptionally regulate the expression of EphB4. Downregulation of HoxA9 reduced the expression of EphB4. Chromatin immunoprecipitation revealed that HoxA9 interacted with the EphB4 promoter, whereas a deletion construct of HoxA9 without DNA-binding motif (Delta(aa) 206 272) did not bind. Consistently, HoxA9 wild-type overexpression activated the EphB4 promoter as determined by reporter gene expression. HoxA9 binds to the EphB4 promoter and stimulates its expression resulting in an increase of endothelial cell migration and tube forming activity. Thus, modulation of EphB4 expression may contribute to the proangiogenic effect of HoxA9 in endothelial cells. PMID- 14764453 TI - LR11, an LDL receptor gene family member, is a novel regulator of smooth muscle cell migration. AB - LR11, a member of the LDL receptor family, is highly expressed in vascular smooth muscle cells (SMCs) of the hyperplastic intima, and induces enhanced migration of SMCs in vitro via its upregulation of urokinase-type plasminogen activator receptor (uPAR) expression. In this study, we have delineated the mechanism by which LR11 elevates the expression levels of uPAR in SMCs. Secretion of soluble LR11 is induced in SMCs during the rapidly proliferating phase, and the secreted LR11 induces the migration activities of SMCs. Both the cell-anchored and secreted forms of LR11 have the capacity to bind to and form complexes with uPAR. LR11-overexpressing cells show significantly enhanced uPAR binding, but decreased uPAR internalization. LR11 colocalizes with uPAR on the cell surface and inhibits the LDL receptor-related protein (LRP)-mediated binding and internalization of uPAR. Thus, LR11 mediates the uPAR localization to the plasma membrane. LR11 is highly expressed in the atheromatous plaque areas of apoE knockout mice, particularly in the intimal SMCs at the border between intima and media. The neutralization of LR11 function with anti-LR11 antibody reduced cuff-induced intimal thickness in mice. The novel mechanism of regulation of uPAR localization in SMCs accompanied with enhanced migration activity possibly constitutes an important factor in the process of atherosclerosis and arterial remodeling. PMID- 14764455 TI - The Janus faces of NO? PMID- 14764454 TI - Regeneration of the infarcted heart with stem cells derived by nuclear transplantation. AB - Nuclear transfer techniques have been proposed as a strategy for generating an unlimited supply of rejuvenated and histocompatible stem cells for the treatment of cardiac diseases. For this purpose, c-kit-positive fetal liver stem cells obtained from cloned embryos were injected in the border zone of infarcted mice to induce tissue reconstitution. Cloned embryos were derived from somatic cell fusion between nuclei of cultured LacZ-positive fibroblasts and enucleated oocytes of a different mouse strain. We report that regenerating myocardium replaced 38% of the scar at 1 month. The rebuilt tissue expressed LacZ and was composed of myocytes and vessels connected with the coronary circulation. Myocytes were functionally competent and expressed contractile proteins, desmin, connexin43, and N-cadherin. These structural characteristics indicated that the new myocytes were electrically and mechanically coupled. Similarly, the formed coronary arterioles and capillary structures contained blood and contributed, therefore, to tissue oxygenation. Cardiac replacement resulted in an improvement of ventricular hemodynamics and in a reduction of diastolic wall stress. These beneficial effects were obtained by stem cell transdifferentiation and commitment to the cardiac cell lineages. Myocardial growth was independent from fusion of the injected stem cells with preexisting partner cells. In conclusion, c-kit positive stem cells derived by nuclear transfer cloning restore infarcted myocardium. Although problems currently plague nuclear transplantation, including the potential for epigenetic and imprinting abnormalities, stem cells derived from cloned embryos are sufficiently normal to repair damaged tissue in vivo. Importantly, the magnitude of myocardial regeneration obtained in this study is significantly superior to that achieved with adult bone marrow cells. PMID- 14764456 TI - Over-expression of gastrin-releasing peptide in human esophageal squamous cell carcinomas. AB - Gastrin-releasing peptide (GRP) is known as an autocrine growth factor for a number of gastrointestinal cancers. There is, however, little information on the expression of GRP in the squamous epithelia and squamous cell carcinoma, particularly in the esophagus. With a differential display approach, up-regulated GRP was observed in human esophageal squamous cell carcinoma (ESCC) samples obtained from a high-risk area for esophageal cancer, Linzhou in northern China. Up-regulation of phosphoglycerate mutase and P311 HUM (3.1) and down-regulation of keratin 13, cystatin B, endoglin and annexin I were observed. Using a reverse transcription-polymerase chain reaction (RT-PCR) method, significant over expression of GRP was observed in 10 out of 12 ESCC samples (83.3%) and all four ESCC cell lines. With in situ hybridization, GRP mRNA expression was detected in nine out of 21 (42.8%) samples with basal cell hyperplasia (BCH), five out of seven (71.4%) samples with dysplasia (DYS) and 17 out of 24 (70.9%) ESCC samples. In contrast, GRP was expressed only in three out of 16 (18.7%) normal epithelium. Digital image analysis revealed that the mean value of GRP expression index, determined by intensity and area ratio of staining, was 0.19 in normal epithelium, 1.23 in BCH, 2.94 in DYS and 2.38 in ESCC, showing a progressive increase. Studies on ESCC cell lines showed GRP increased cell growth in a dose dependent pattern in GRP receptor-positive ESCC cells, but not in GRP receptor negative ESCC cells. GRP (1 mM) also increased cyclooxygenase-2 protein expression by 3.4-fold. This is the first demonstration that GRP is over expressed in ESCC, and its over-expression may play a role in ESCC development and growth. PMID- 14764457 TI - p53 Protein interacts specifically with the meiosis-specific mammalian RecA-like protein DMC1 in meiosis. AB - The tumor suppressor protein p53 is specifically expressed during meiosis in spermatocytes. Subsets of p53 knockout mice exhibit testicular giant cell degenerative syndrome, which suggests p53 may be associated with meiotic cell cycle and/or DNA metabolism. Here, we show that p53 binds to the mouse meiosis specific RecA-like protein Mus musculus DMC1 (MmDMC1). The C-terminal domain (amino acid 234-340) of MmDMC1 binds to DNA-binding domain of p53 protein. p53 might be involved in homologous recombination and/or checkpoint function by directly binding to DMC1 protein to repress genomic instability in meiotic germ cells. PMID- 14764458 TI - Cytokine production by alveolar macrophages is down regulated by the alpha methylhydroxylation pathway of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK). AB - NNK, a nicotine-derived nitrosamine, is a potent lung carcinogen that generates electrophilic intermediates capable of damaging DNA. The effects of NNK on the immune response, which may facilitate lung carcinogenesis, are poorly understood. Alveolar macrophages (AM), a key cell in the maintenance of lung homeostasis, metabolize NNK via two major metabolic activation pathways: alpha methylhydroxylation and alpha-methylenehydroxylation. We have shown previously that NNK inhibits the production of interleukin-12 (IL-12) and tumor necrosis factor (TNF), but stimulates the production of IL-10 and prostaglandin E(2) (PGE(2)) by AM. In the present study, we investigated the contribution of each activation pathway in the modulation of AM function. We used two precursors, 4 [(acetoxymethyl)-nitrosamino]-1-(3-pyridyl)-1-butanone (NNKOAc) and N nitro(acetoxymethyl)methylamine (NDMAOAc), which generate the reactive electrophilic intermediates [4-(3-pyridyl)-4-oxo-butanediazohydroxide and methanediazohydroxide, respectively] in high yield and exclusively. Rat AM cell line, NR8383, was stimulated and treated with different concentrations of NNKOAc or NDMAOAc (12, 25 and 50 microM). Mediator release was measured in cell-free supernatants. NNKOAc significantly inhibited the production of IL-10, IL-12, TNF and nitric oxide but increased the release of PGE(2) and cyclooxygenase-2 expression suggesting that the alpha-methylhydroxylation pathway might be responsible for NNK modulation of AM cytokine release. In contrast, NDMAOAc did not modulate AM mediator production. However, none of these precursors, alone or in combination, could explain the stimulation of AM IL-10 production by NNK. Our results suggest that the alpha-methylhydroxylation of NNK leading to DNA pyridyloxobutylation also modulates cytokine production in NNK-treated AM. PMID- 14764459 TI - Decrease of mitochondrial DNA content and energy metabolism in renal cell carcinoma. AB - To elucidate the relationship between tumor genesis and the mitochondrial energy metabolism in renal neoplasms, we studied three individual enzyme activities of the oxidative phosphorylation, two components of the Krebs cycle and the mitochondrial DNA content of renal carcinomas including 29 conventional, five papillary, two unclassified carcinomas with sarcomatoid features and one collecting duct carcinoma. A significant reduction of all mitochondrial enzyme activities including complex V, as well as of the mitochondrial DNA content was detected in 34 of 37 renal carcinoma tissues as compared with control kidney. Mitochondrial enzyme activities and mitochondrial DNA levels were not statistically different between the conventional, papillary and unclassified sarcomatoid type of renal carcinoma and did not correlate with tumour grade, metastasis, ploidy and proliferative activity as determined by Ki-67 staining. Taken together, our data indicate that a co-ordinated down-regulation of all components necessary for mitochondrial energy metabolism occurs in most renal carcinomas as an early event in carcinoma formation, which does not change with progression of the disease. PMID- 14764460 TI - Promotion of carcinogenesis and oxidative stress by dietary cholesterol in rat prostate. AB - The association between prostate cancer risk and dietary fat consumption is well documented and explained partly by accelerated lipid peroxidation. We explored the possible effects of high dietary cholesterol on carcinogenesis and oxidative stress in the prostate of ACI/Seg rats. The rats develop prostate cancer spontaneously late in the life, providing an appropriate model to explore prolonged dietary conditions. Two groups of 20-week-old male rats, 28 each, were fed either a basal diet or a basal diet supplemented with 1% cholesterol (high cholesterol diet), and killed at 100 weeks of age. Rats on the high cholesterol diet developed adenocarcinoma in the ventral prostate more frequently (26 versus 4%, P = 0.023). In the repeat study, 26 rats each were treated similarly and killed at 80 weeks for histology and oxidative stress assay. Oxidative stress was assessed by measuring the plasma and intra-prostatic levels of vitamin E, vitamin C, uric acid and the oxidized and reduced forms of coenzyme Q(9). The relative amount of oxidized form of coenzyme Q(9) is a sensitive marker of oxidative stress. Rats on the high cholesterol diet demonstrated a higher incidence of atypical prostatic hyperplasia (24 versus 4%, P = 0.049). Also, the prostate showed a 2-fold increase (203% of the control) in the relative amounts of the oxidized form of coenzyme Q(9) and reciprocal reduction of vitamin C (9.5% of the control) and uric acid (46% of the control) levels (P < 0.01), with a minimal change in vitamin E. The plasma levels of these compounds were not affected by dietary conditions. These results indicated that long-term feeding of a 1% cholesterol diet promoted carcinogenesis and tissue oxidative stress in rat prostate. The role of dietary fat and oxidative stress in prostate carcinogenesis needs further investigation. PMID- 14764461 TI - Ribozyme-mediated down-regulation of survivin expression sensitizes human melanoma cells to topotecan in vitro and in vivo. AB - The ability of melanoma cells to evade engagement of apoptosis plays a significant role in their resistance to chemotherapy. In an attempt to lower the apoptotic threshold of melanoma cells as a possible strategy to increase their drug sensitivity, we generated a hammerhead ribozyme to down-regulate the expression of the anti-apoptotic protein survivin. The JR8 human melanoma cell line was stably transfected with the active ribozyme RZsurv (targeting the 3' end of the GUC294 triplet in the exon 3 of the survivin mRNA) or the catalytically inactive ribozyme mutRZsurv (carrying a mutation in the catalytic core of RZsurv). Two polyclonal cell populations expressing the active (JR8/RZsurv) or the mutant (JR8/mutRZsurv) ribozyme were selected for the study. JR8/RZsurv cells were characterized by a markedly lower survivin protein level than JR8 parental cells, whereas a negligible reduction in survivin expression was observed in JR8/mutRZsurv cells. JR8/RZsurv cells showed a significantly increased sensitivity to the topoisomerase-I inhibitor topotecan (as detected by clonogenic cell survival) compared with JR8/mutRZsurv cells. Moreover, the extent of drug induced apoptosis (in terms of percentage of apoptotic nuclei and level of caspase-9 and caspase-3 catalytic activity) was significantly greater in JR8/RZsurv than in JR8/mutRZsurv cells. Finally, an increased antitumor activity of oral topotecan was observed in JR8/RZsurv cells grown as xenograft tumors in athymic nude mice compared with JR8/mutRZsurv cells. These results demonstrate that attenuation of survivin expression renders human melanoma cells more susceptible to topotecan-induced apoptosis and more responsive to in vivo treatment, and support the concept that survivin is an attractive target for new therapeutic interventions in melanoma. PMID- 14764462 TI - Secondary prevention for stroke and transient ischaemic attacks. PMID- 14764463 TI - Compulsory screening of immigrants for tuberculosis and HIV. PMID- 14764464 TI - Portable echocardiography. PMID- 14764465 TI - Screening without evidence of efficacy. PMID- 14764466 TI - FDA advisory panel calls for suicide warnings over new antidepressants. PMID- 14764467 TI - British cancer death rates fell by 12% between 1972 and 2002. PMID- 14764468 TI - Patients can be made to have HIV test to protect doctor. PMID- 14764469 TI - Quebec will make doctors disclose their HIV status. PMID- 14764470 TI - Women are being let down in efforts to stem HIV/AIDS. PMID- 14764474 TI - Research bodies disappointed by decision to cancel primate research laboratory. PMID- 14764475 TI - Traditional rulers in northern Nigeria call for halt to polio vaccination. PMID- 14764476 TI - Drug company targets US state health officials. PMID- 14764479 TI - Spanish drug editor wins case brought by Merck, Sharp Dohme. PMID- 14764480 TI - Ethnic minority groups criticise US report on healthcare disparities. PMID- 14764481 TI - WHO investigates possible human to human transmission of avian flu. PMID- 14764482 TI - Doing as well as thinking. PMID- 14764483 TI - Could joining EU club spell disaster for the new members? PMID- 14764484 TI - Europe plans to standardise rules on tissue engineering. PMID- 14764486 TI - Teenager seeks to overturn GMC judgment. PMID- 14764488 TI - GMC decision on Egyptian doctor overturned by High Court. PMID- 14764489 TI - Poliovirus spreads beyond Nigeria after vaccine uptake drops. PMID- 14764491 TI - UN criticises developed countries for reneging on promises on sexual health. PMID- 14764492 TI - Population based intervention to change back pain beliefs: three year follow up population survey. PMID- 14764493 TI - Undiagnosed coeliac disease at age seven: population based prospective birth cohort study. PMID- 14764494 TI - Use of tobacco products as dentifrice among adolescents in India: questionnaire study. PMID- 14764495 TI - Blood pressure, haemorrhagic stroke, and ischaemic stroke: the Korean national prospective occupational cohort study. PMID- 14764496 TI - Scottish general practitioners' willingness to take part in a post-retirement retention scheme: questionnaire survey. PMID- 14764497 TI - Persistent crying in babies. PMID- 14764498 TI - Investigating sudden unexpected deaths in infancy and childhood and caring for bereaved families: an integrated multiagency approach. PMID- 14764499 TI - Severe hypocalcaemia after being given intravenous bisphosphonate. PMID- 14764501 TI - Assessment of dependence and motivation to stop smoking. PMID- 14764502 TI - New providers in UK health care. PMID- 14764503 TI - Changing clinical behaviour by making guidelines specific. PMID- 14764504 TI - Long term mortality after starvation during the Leningrad siege: crucial confounding factor was overlooked. PMID- 14764505 TI - Long term mortality after starvation during the Leningrad siege: no evidence that starvation around puberty causes later cardiovascular disease. PMID- 14764506 TI - Critical care outreach team's effect on patient outcome: more information is needed. PMID- 14764507 TI - Critical care outreach team's effect on patient outcome: other conclusions are possible. PMID- 14764508 TI - Treatment of unilateral visual impairment on preschool vision screening: study leaves questions unanswered. PMID- 14764509 TI - Treatment of unilateral visual impairment on preschool vision screening: mild amblyopia should still be treated. PMID- 14764510 TI - Treatment of unilateral visual impairment on preschool vision screening: preschool vision screening should continue, perhaps earlier. PMID- 14764511 TI - Volunteer counsellors for supporting breast feeding: support for breast feeding must be proactive. PMID- 14764512 TI - Low prices for HIV drugs are only one step forward. PMID- 14764513 TI - American medicine meets the American dream: moral dimension is important. PMID- 14764514 TI - American medicine meets the American dream: much inequality is self perpetuated. PMID- 14764515 TI - Volunteer counsellors for supporting breast feeding: peer support is worth further investigation. PMID- 14764516 TI - Older people and the "patient centred" NHS: prioritisation will be for financial reasons rather than by need. PMID- 14764517 TI - Older people and the "patient centred" NHS: proactive support in primary care can avert admissions. PMID- 14764519 TI - Older people and the "patient centred" NHS: choice for mentally ill patients will be even less with new legislation. PMID- 14764520 TI - Substitution of the gamma-chain Asn308 disturbs the D:D interface affecting fibrin polymerization, fibrinopeptide B release, and FXIIIa-catalyzed cross linking. AB - Crystallographic structures indicate that gamma-chain residue Asn308 participates in D:D interactions and indeed substitutions of gammaAsn308 with lysine or isoleucine have been identified in dysfibrinogens with impaired polymerization. To probe the role of Asn308 in polymerization, we synthesized 3 variant fibrinogens: gammaAsn308 changed to lysine (gammaN308K), isoleucine (gammaN308I), and alanine (gammaN308A). We measured thrombin-catalyzed polymerization by turbidity, fibrinopeptide release by high-performance liquid chromatography, and factor XIIIa-catalyzed cross-linking by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. In the absence of added calcium, polymerization was clearly impaired with all 3 variants. In contrast, at 0.1 mM calcium, only polymerization of gammaN308K remained markedly abnormal. The release of thrombin-catalyzed fibrinopeptide B (FpB) was delayed in the absence of calcium, whereas at 1 mM calcium FpB release was delayed only with gammaN308K. Factor XIIIa-catalyzed gamma-gamma dimer formation was delayed with fibrinogen (in absence of thrombin), whereas with fibrin (in presence of thrombin) gamma-gamma dimer formation of only gammaN308K was delayed. These data corroborate the recognized link between FpB release and polymerization. They show fibrin cross-link formation likely depends on the structure of protofibrils. Together, our results show substitution of Asn308 with a hydrophobic residue altered neither polymer formation nor polymer structure at physiologic calcium concentrations, whereas substitution with lysine altered both. PMID- 14764521 TI - Dynamics of cytokine expression in HIV productively infected primary CD4+ T cells. AB - Using intracellular p24 staining to discriminate between bystander and HIV productively infected cells, we evaluated the properties of HIV productively infected cells in terms of cytokine expression, activation status, apoptosis, and cell proliferation. We demonstrate that HIV productively infected primary CD4(+) T cells express 12- to 47-fold higher type 1 cytokines than bystander or mock infected cells. The frequency of HIV productive replication occurred predominantly in T-helper 1 (Th1), followed by Th0, then by Th2 cells. These productively infected cells expressed elevated levels of CD95, CD25, CXC chemokine receptor 4 (CXCR4), and CC chemokine receptor 5 (CCR5). While productively infected cells were only 1.8-fold higher in apoptosis frequency, they up-regulated the antiapoptotic protein B-cell leukemia 2 (Bcl-2) by 10-fold. Up-regulation of interleukin-2 (IL-2) and Bcl-2 were dependent on phosphatidylinositol-3-kinase signal transduction, given that it was down regulated by Wortmanin treatment. Additionally, 60% of productively infected cells entered the cell cycle, as evaluated by Ki67 staining, but none divided, as evaluated by carboxyfluoresccin diacetate succinimidyl ester (CFSE) staining. Evaluation of cell cycle progression by costaining for DNA and RNA indicated that the cells were arrested in G(2)/M. Collectively, these data indicate that HIV replication occurs predominantly in Th1 cells and is associated with immune activation and up-regulation of Bcl-2, conferring a considerable degree of protection against apoptosis in the productively infected subpopulation. PMID- 14764522 TI - T lymphocytes of recipient origin may contribute to the recovery of specific immune response toward viruses and fungi in children undergoing cord blood transplantation. AB - Patients undergoing allogeneic cord blood transplantation (CBT) benefit from a low risk of graft-versus-host disease (GVHD), but there are still concerns that they be able to recover an effective immune capacity early after transplantation. We investigated the ability to develop in vitro T-lymphocyte-mediated immune response toward human cytomegalovirus and Candida albicans antigens, early and late after transplantation, in children given cord blood transplants from either an HLA-identical sibling or an unrelated donor. Proliferative capacity and frequency of antigen-specific T cells were evaluated; antigen-specific CD4(+) T cell clones were also generated and characterized for T-cell receptor repertoire diversity, cytokine phenotype, and their origin (either from donor or patient). We found that the majority of recipients can develop a specific response to viral or fungal antigens already early after transplantation. Antigen-specific T-cell clones of both donor and recipient origin contributed to the reconstitution of immune response. Antigen-specific T lymphocytes of recipient origin were detected in patients receiving a transplant from a relative, after a chemotherapy-based conditioning regimen, and who did not have GVHD. Our results document, at a clonal level, that after CBT recovery of either polyclonal or pauciclonal T-cell response toward widespread pathogens is prompt, with some patients benefiting from a contribution of recipient-derived cells. PMID- 14764524 TI - Transgenic mice studies demonstrate a role for platelet factor 4 in thrombosis: dissociation between anticoagulant and antithrombotic effect of heparin. AB - The platelet-specific chemokine platelet factor 4 (PF4) is released in large amounts at sites of vascular injury. PF4 binds to heparin with high affinity, but its in vivo biologic role has not been defined. We studied the role of PF4 in thrombosis using heterozygote and homozygote PF4 knock-out mice (mPF4(+/-) and mPF4(-/-), respectively) and transgenic mice overexpressing human PF4 (hPF4(+)). None of these lines had an overt bleeding diathesis, but in a FeCl(3) carotid artery thrombosis model, all showed impaired thrombus formation. This defect in thrombus formation in the mPF4(-/-) animals was corrected by infusing hPF4 over a narrow concentration range. The thrombotic defect in the mPF4(+/-) and mPF4(-/-) animals was particularly sensitive to infusions of the negatively charged anticoagulant heparin. However, the same amount of heparin paradoxically normalized thrombus formation in the hPF4(+) animals, although these animals were anticoagulated systemically. Upon infusion of the positively charged protein, protamine sulfate, the reverse was observed with mPF4(+/-) and mPF4(-/-) animals having improved thrombosis, with the hPF4(+) animals having worsened thrombus formation. These studies support an important role for PF4 in thrombosis, and show that neutralization of PF4 is an important component of heparin's anticoagulant effect. The mechanisms underlying these observations of PF4 biology and their clinical implications remain to be determined. PMID- 14764523 TI - Clonotypic IgM V/D/J sequence analysis in Waldenstrom macroglobulinemia suggests an unusual B-cell origin and an expansion of polyclonal B cells in peripheral blood. AB - Analysis of clonotypic immunoglobulin M (IgM) from 15 patients with Waldenstrom macroglobulinemia (WM) showed a strong preferential use of the VH3/JH4 gene families. Identification of the WM IgM V/D/J was validated using single-cell analysis, confirming its presence in most B cells. Despite the extensive hypermutated VH genes in 13 of 15 patients, statistical analysis of framework/complementary-determining region (FR/CDR) mutation patterns suggests that they might have escaped antigenic selection. Neither intraclonal diversity nor isotype switching was detectable. Membranous and secreted forms of clonotypic IgM transcripts were present in bone marrow and blood. Single-cell analysis showed that clonotypic B cells coexpress CD20, surface IgM (sIgM), and sIgD but that they lack CD138. Most B cells lacked memory marker CD27 despite their hypermutated variable regions otherwise suggestive of memory status. At diagnosis, circulating B cells in WM are largely clonotypic. However, when monoclonal IgM levels are decreased, clonotypic frequencies are substantially reduced despite elevated CD20+ cells, shown to be polyclonal by DNA sequencing and CDR3 fragment analysis. Thus, WM includes the expansion of circulating, polyclonal B cells. Overall, this work suggests that WM may originate from a largely VH3-restricted, somatically mutated, predominantly CD27(-)IgM(+)IgD+ population that cannot undergo class switching, suggestive of B cells that might have bypassed the germinal center. PMID- 14764525 TI - Subspeciality training in hematology and oncology, 2003: results of a survey of training program directors conducted by the American Society of Hematology. AB - A survey of directors of adult and pediatric hematology/oncology subspecialty training programs in the United States and Canada was conducted to assess the environment in which recruitment and training is conducted in these medical disciplines. A total of 107 program directors responded to the survey, representing 66% of internal medicine and 47% of pediatric subspecialty programs in hematology or hematology/oncology. Specific areas covered in the web-based questionnaire included the type and demographics of the training program, profile of the training program director, characteristics of the applicant pool and existing trainee recruits, characteristics of the training program environment and curricula, research productivity of trainees, and the career pathways taken by recent training program graduates (including dominant areas of clinical interest). The results of this survey show considerable heterogeneity in the recruiting practices and the environment in which subspecialty training occurs, leading the authors to recommend improvements in or a heightened attention to issues, including recruitment of minority trainees, flexibility to recruit international medical school graduates, timing of trainee acceptance, maintaining the financial support of Medicare graduation medical education (GME), training of physician scientists, organization of the continuity clinic experience, visibility of nonmalignant hematology as a career path, and level of training program director support. PMID- 14764526 TI - Neutrophils in Barth syndrome (BTHS) avidly bind annexin-V in the absence of apoptosis. AB - Barth syndrome (BTHS) is a rare X-linked disease characterized by a triad of dilated cardiomyopathy, skeletal myopathy, and neutropenia. The disease is associated with mutations of the TAZ gene, resulting in defective cardiolipin (CL), an important inner mitochondrial membrane component. Untreated boys die in infancy or early childhood from septicemia or cardiac failure. To date, neutrophil function has never been studied. Directed motility and killing activity of neutrophils was investigated in 7 BTHS patients and found normal in those tested. The circulating neutrophils and eosinophils (but not monocytes or lymphocytes) showed annexin-V binding, suggesting phosphatidylserine (PS) exposure due to apoptosis. However, caspase activity was absent in fresh BTHS cells. Unexpectedly, the near absence of CL impacted neither the mitochondrial mass and shape in fresh BTHS neutrophils nor mitochondrial clustering and Bax translocation upon apoptosis. Annexin-V binding to BTHS neutrophils was not caused by phospholipid scrambling. Moreover, freshly purified BTHS neutrophils were not phagocytosed by macrophages. In sum, a massive number of circulating annexin-V-binding neutrophils in the absence of apoptosis can be demonstrated in BTHS. These neutrophils expose an alternative substrate for annexin-V different from PS and not recognized by macrophages, excluding early clearance as an explanation for the neutropenia. PMID- 14764527 TI - Survival of children with sickle cell disease. AB - Contemporary survival data are not available for children with sickle cell disease (SCD). The few previous childhood SCD cohort studies do not reflect the benefits of modern therapy. We defined an inception cohort of newborns with sickle cell anemia (SS), sickle-beta degrees -thalassemia (S beta degrees ), sickle-hemoglobin C disease (SC), or sickle-beta(+)-thalassemia (Sbeta(+)) who were identified by newborn screening and followed for up to 18 years. The incidence of death and stroke were calculated. Overall survival, SCD-related survival (considering only SCD-related deaths), and strokefree survival were determined. The 711 subjects provided 5648 patient-years of observation. Twenty five subjects died; mean age at death was 5.6 years. Five patients died from infection. Thirty had at least one stroke. Among SS and Sbeta degrees subjects (n = 448), the overall rates of death and stroke were 0.59 and 0.85/100 patient years. Survival analysis of SS and Sbeta degrees subjects predicted the cumulative overall, SCD-related, and stroke-free survival to be 85.6%, 93.6%, and 88.5% by 18 years of age. No SCD-related deaths or strokes occurred in SC or Sbeta(+) subjects (n = 263). Childhood mortality from SCD is decreasing, the mean age at death is increasing, and a smaller proportion of deaths are from infection. PMID- 14764528 TI - Familial essential thrombocythemia associated with a dominant-positive activating mutation of the c-MPL gene, which encodes for the receptor for thrombopoietin. AB - One Japanese pedigree of familial essential thrombocythemia (FET) inherited in an autosomal-dominant manner is presented. A unique point mutation, serine 505 to asparagine 505 (Ser505Asn), was identified in the transmembrane domain of the c MPL gene in all of the 8 members with thrombocythemia, but in none of the other 8 unaffected members in this FET family. The Ba/F3 cells expressing the mutant Asn505 acquired interleukin 3 (IL-3)-independent survival capacity, whereas those expressing wild-type Ser505 did not. The autonomous phosphorylation of Mek1/2 and Stat5b was observed in the mutant Ba/F3 cells in the absence of IL-3. The former was also found in platelets derived from the affected individual in the absence of thrombopoietin. These results show that the Asn505 is an activating mutation with respect to the intracellular signaling and survival of the cells. This is the first report of FET deriving from a dominant-positive activating mutation of the c-MPL gene. PMID- 14764529 TI - Comparative proteomics of primitive hematopoietic cell populations reveals differences in expression of proteins regulating motility. AB - Lineage-marker depleted (Lin(-)) murine bone marrow cells expressing stem cell antigen 1 (Sca-1) were sorted on the basis of stem cell factor receptor (c-kit) expression to obtain Lin(-)Sca(+)Kit(+) or Lin(-)Sca(+)Kit(-) cells. Lin( )Sca(+)Kit(-) cells have a markedly greater chemotactic response to stromal derived factor-1 (SDF-1). Using a novel fluorescent stain, we show that both populations generate similar levels of a key messenger, phosphatidylinositol 3,4,5 trisphosphate (PIP(3)), in response to SDF-1. Differences in motile behavior may therefore lie downstream of phosphatidylinositol 3-kinase (PI3 kinase) activation at the level of cytoskeleton regulation. The 2 cell populations were compared using 2-dimensional difference gel electrophoresis (2D DIGE), with a maleimide CyDye fluorescent protein labeling technique that has enhanced sensitivity for low abundance samples. Comparative proteomic analysis of Cy3- and Cy5-labeled protein samples allows relative quantification of protein spots present in both cell populations; of these, 73% were common. Key protein differences were adseverin and gelsolin, actin micro-filament splicing proteins, regulated by Rac, downstream of PI3-kinase activation. Adseverin was shown to be acetylated, a novel modification for this protein. Differences in major regulators of cell shape and motility between the 2 populations can explain the differential response to SDF-1. PMID- 14764530 TI - Severe chronic graft-versus-host disease is characterized by a preponderance of CD4(+) effector memory cells relative to central memory cells. AB - Donor alloreactive CD4(+) T cells are important to the pathogenesis of chronic graft-versus-host disease (cGVHD), but specific subsets of CD4(+) T cells responsible for GVHD have not been defined. We hypothesized that cGVHD might be associated with a preponderance of CD4(+) effector memory cells (CCR7( )/CD62L(low), CD4(EM)). We analyzed CCR7 and CD62L expression on CD4(+) T cells from stem cell transplantation patients, who did or did not develop cGVHD, and healthy donors. Patients with cGVHD had a higher percentage of CD4(EM) cells (35.5% +/- 2.9%) than healthy donors (13.8% +/- 0.7%; P <.0001) or patients without cGVHD that received a transplant (21.7% +/- 2.1%; P <.01). Using corticosteroid dose as a surrogate marker for cGVHD severity, severe cGVHD was associated with a higher percentage of CD4(EM) cells. The proportion of CD4(EM) cells in corticosteroid-dependent patients with systemic lupus erythematosis or Wegener granulomatosis did not differ from patients without cGVHD that received a transplant. This finding implies that overrepresentation of CD4(EM) cells is a unique feature of cGVHD. PMID- 14764531 TI - Regulatory elements of the EKLF gene that direct erythroid cell-specific expression during mammalian development. AB - Erythroid Kruppel-like factor (EKLF) plays an essential role in enabling beta globin expression during erythroid ontogeny. It is first expressed in the extraembryonic mesoderm of the yolk sac within the morphologically unique cells that give rise to the blood islands, and then later within the hepatic primordia. The BMP4/Smad pathway plays a critical role in the induction of EKLF, and transient transfection analyses demonstrate that sequences located within less than 1 kb of its transcription initiation site are sufficient for high-level erythroid-specific transcription. We have used transgenic analyses to verify that 950 bp located adjacent to the EKLF start site of transcription is sufficient to generate lacZ expression within the blood islands as well as the fetal liver during embryonic development. Of particular importance are 3 regions, 2 of which overlap endogenous erythroid-specific DNase hypersensitive sites, and 1 of which includes the proximal promoter region. The onset of transgene expression mimics that of endogenous EKLF as it begins by day 7.5 (d7.5) to d8.0. In addition, it exhibits a strict hematopoietic specificity, localized only to these cells and not to the adjacent vasculature at all stages examined. Finally, expression is heterocellular, implying that although these elements are sufficient for tissue specific expression, they do not shield against the position effects of adjacent chromatin. These analyses demonstrate that a surprisingly small DNA segment contains all the information needed to target a linked gene to the hematopoietic compartment at both early and later stages of development, and may be a useful cassette for this purpose. PMID- 14764532 TI - Dendritic cell-activated CD44hiCD8+ T cells are defective in mediating acute graft-versus-host disease but retain graft-versus-leukemia activity. AB - Graft versus host disease (GVHD) is triggered by host antigen-presenting cells (APCs) that activate donor T cells to proliferate and differentiate, but which APC-activated donor T-cell subsets mediate GVHD versus beneficial antitumor effects is not known. Using a CD8(+) T cell-dependent mouse model of human GVHD, we found that host dendritic cell (DC)-induced CD44(hi)CD8(+) effector/memory T cells were functionally defective in inducing GVHD, whereas CD44(lo)CD8(+) naive phenotype T cells were extremely potent GVHD inducers. Depletion of CD44(lo)CD8(+) T cells from host DC-stimulated T cells before transplantation prevented GVHD without impairing their antitumor activity in vivo. Compared with CD44(lo)CD8(+) T cells, CD44(hi)CD8(+) T cells expressed high levels of Fas and were efficiently deleted in vivo following transplantation. These results suggest that ex vivo allogeneic DC stimulation of donor CD8(+) T cells may be useful for the prevention of GVHD and for optimizing antitumor therapies in vivo. PMID- 14764533 TI - Pim-1 kinase inhibits STAT5-dependent transcription via its interactions with SOCS1 and SOCS3. AB - Signal transducer and activator of transcription 5 (STAT5) plays a critical role in cytokine-induced survival of hematopoietic cells. One of the STAT5 target genes is pim-1, which encodes an oncogenic serine/threonine kinase. Here we demonstrate that Pim-1 inhibits STAT5-dependent transcription in cells responsive to interleukin-3, prolactin, or erythropoietin. Ectopic expression of Pim-1 in cytokine-dependent FDCP1 myeloid cells results in reduced tyrosine phosphorylation and DNA binding of STAT5, indicating that Pim-1 interferes already with the initial steps of STAT5 activation. However, the Pim-1 kinase does not directly phosphorylate or bind to STAT5. By contrast, Pim-1 interacts with suppressor of cytokine signaling 1 (SOCS1) and SOCS3 and potentiates their inhibitory effects on STAT5, most likely via phosphorylation-mediated stabilization of the SOCS proteins. Thus, both Pim and SOCS family proteins may be components of a negative feedback mechanism that allows STAT5 to attenuate its own activity. PMID- 14764534 TI - Combined deficiency in IkappaBalpha and IkappaBepsilon reveals a critical window of NF-kappaB activity in natural killer cell differentiation. AB - Nuclear factor kappaB (NF-kappaB) transcription factors are key regulators of immune, inflammatory, and acute-phase responses and are also implicated in the control of cell proliferation and apoptosis. While perturbations in NF-kappaB activity impact strongly on B- and T-cell development, little is known about the role for NF-kappaB in natural killer (NK) cell differentiation. Inhibitors of NF kappaB (IkappaBs) act to restrain NF-kappaB activation. We analyzed the cell intrinsic effects of deficiencies in 2 IkappaB members (IkappaBalpha and IkappaBepsilon) on NK cell differentiation. Neither IkappaBalpha nor IkappaBepsilon deficiency had major effects on NK cell generation, while their combined absence led to NF-kappaB hyperactivation, resulting in reduced NK cell numbers, incomplete NK cell maturation, and defective interferon gamma (IFN gamma) production. Complementary analysis of transgenic mice expressing an NF kappaB-responsive reporter gene showed increased NF-kappaB activity at the stage of NK cell development corresponding to the partial block observed in IkappaBalpha x IkappaBepsilon-deficient mice. These results define a critical window in NK cell development in which NF-kappaB levels may be tightly controlled. PMID- 14764535 TI - Zileuton induces hemoglobin F synthesis in erythroid progenitors: role of the L arginine-nitric oxide signaling pathway. AB - Induction of fetal hemoglobin (Hb F) is an important therapeutic tool in ameliorating complications of sickle cell disease. Nitric oxide has been implicated in the mechanism of Hb F synthesis induced by hydroxyurea (HU). This study examined whether zileuton (ZL), a structural analog of hydroxyurea, possessed Hb F-inducing properties and the potential role nitric oxide plays. ZL caused a dose-dependent increase in gamma-globin expression in K562 cells. This effect was confirmed by a dose-dependent increase in Hb F synthesis in erythroid progenitors from individuals with sickle cell anemia and normal hemoglobin genotypes. l-arginine had no effect on Hb F production; however, it dose dependently inhibited ZL's ability to induce Hb F. The nitric oxide synthase inhibitor N(G)-monomethyl-l-arginine (l-NMMA) inhibited l-arginine's effect and restored ZL-mediated increase in Hb F synthesis. In addition, 8-PCPT-cGMP (8-(4 chlorophenylthio)guanosine 3',5'-cyclic monophosphate) inhibited ZL-mediated induction of Hb F synthesis. When comparing l-NMMA effects alone on ZL and HU, a partial reversal of increased Hb F synthesis was seen only with HU. Neither l arginine alone nor l-arginine in combination with l-NMMA effected hydroxyurea mediated induction of Hb F synthesis. This study demonstrates that ZL induces Hb F through a mechanism that involves l-arginine/nitric oxide/cGMP in a manner distinctly different from HU. PMID- 14764536 TI - Characterization of childhood acute lymphoblastic leukemia xenograft models for the preclinical evaluation of new therapies. AB - Continuous xenografts from 10 children with acute lymphoblastic leukemia (ALL) were established in nonobese diabetic/severe combined immunodeficient (NOD/SCID) mice. Relative to primary engrafted cells, negligible changes in growth rates and immunophenotype were observed at second and third passage. Analysis of clonal antigen receptor gene rearrangements in 2 xenografts from patients at diagnosis showed that the pattern of clonal variation observed following tertiary transplantation in mice exactly reflected that in bone marrow samples at the time of clinical relapse. Patients experienced diverse treatment outcomes, including 5 who died of disease (median, 13 months; range, 11-76 months, from date of diagnosis), and 5 who remain alive (median, 103 months; range, 56-131 months, following diagnosis). When stratified according to patient outcome, the in vivo sensitivity of xenografts to vincristine and dexamethasone, but not methotrexate, differed significantly (P =.028, P =.029, and P =.56, respectively). The in vitro sensitivity of xenografts to dexamethasone, but not vincristine, correlated significantly with in vivo responses and patient outcome. This study shows, for the first time, that the biologic and genetic characteristics, and patterns of chemosensitivity, of childhood ALL xenografts accurately reflect the clinical disease. As such, they provide powerful experimental models to prioritize new therapeutic strategies for future clinical trials. PMID- 14764537 TI - VEGF-mediated endothelial P-selectin translocation: role of VEGF receptors and endogenous PAF synthesis. AB - The acute increase in vascular permeability produced by vascular endothelial growth factor (VEGF-A(165)) requires activation of endothelial Flk-1 receptors (VEGFR-2) and stimulation of platelet-activating factor (PAF) synthesis. Like PAF, VEGF-A(165) promotes translocation of P-selectin to the endothelial cell (EC) surface. However, the mechanisms involved remain unknown. By treating human umbilical vein endothelial cells (HUVECs) with VEGF analogs, we show that activation of VEGFR-1 or VEGFR-2 or both induced a rapid and transient translocation of endothelial P-selectin and neutrophil adhesion to activated ECs. The effects mediated by VEGF-A(165) and VEGF-A(121) (VEGFR-1/VEGFR-2 agonists) were blocked by a selective VEGFR-2 inhibitor, SU1498. VEGF-A(165) was twice as potent as VEGF-A(121), which can be explained by the binding capacity of VEGF A(165) to its coreceptor neuropilin-1 (NRP-1). Indeed, treatment with NRP-1 antagonist (GST-Ex7) reduced the effect of VEGF-A(165) to the levels observed upon stimulation with VEGF-A(121). Finally, the use of selective PAF receptor antagonists reduced VEGF-A(165)-mediated P-selectin translocation. Together, these data show that maximal P-selectin translocation and subsequent neutrophil adhesion was mediated by VEGF-A(165) on the activation of VEGFR-2/NRP-1 complex and required PAF synthesis. PMID- 14764538 TI - Chimeric CD19 antibody mediates cytotoxic activity against leukemic blasts with effector cells from pediatric patients who received T-cell-depleted allografts. AB - Relapse is a major problem after transplantation in children with acute B-lineage leukemias, and new therapies are needed to increase graft-versus-leukemia (GvL) effects without inducing graft-versus-host disease (GvHD). Here, we studied the ability of effector cells recovered from patients after transplantation with positive-selected stem cells from alternative donors to induce antibody-dependent cellular cytotoxicity (ADCC). For this purpose, a chimeric CD19 antibody, CD19 4G7chim, was generated. This antibody efficiently mediated ADCC against primary acute lymphoblastic leukemia (ALL) blasts by using purified natural killer (NK) cells from healthy donors or mononuclear cells from patients as effector cells. Increased lysis was obtained after stimulation of effector cells with interleukin 2 (IL-2). ADCC was not prevented by inhibitory effects mediated by HLA class I. We propose that treatment with chimeric CD19 antibodies leading to ADCC by donor derived NK cells may become a therapeutic option for the post-transplantation treatment of minimal residual B-lineage ALLs. PMID- 14764539 TI - Angiopoietin-related growth factor (AGF) promotes angiogenesis. AB - We report here the identification of angiopoietin-related growth factor (AGF) as a positive mediator for angiogenesis. To investigate the biologic function of AGF in angiogenesis, we analyzed the vasculature in the dermis of transgenic mice expressing AGF in mouse epidermal keratinocytes (K14-AGF). K14-AGF transgenic mice were grossly red, especially in the ears and snout, suggesting that hypervascularization had occurred in their skin. Histologic examination of ear skin from K14-AGF transgenic mice revealed increased numbers of microvessels in the dermis, whereas the expression of several angiogenic factors, such as basic fibroblast growth factor (bFGF), vascular endothelial growth factors (VEGFs), and angiopoietin-1 (Ang-1), was decreased. We showed that AGF is a secreted protein and does not bind to tyrosine kinase with immunoglobulin and EGF-homology domain (Tie1) or Tie2 receptors. An in vitro chamber assay revealed that AGF directly promotes chemotactic activity of vascular endothelial cells. Both mouse corneal and matrigel plug assays showed that AGF induces neovascularization in vivo. Furthermore, we found that plasma leakage occurred after direct injection of AGF into the mouse dermis, suggesting that AGF directly induces a permeability change in the local vasculature. On the basis of these observations, we propose that AGF is a novel angiogenic factor and that handling of its biologic functions could lead to novel therapeutic strategies for control of angiogenesis. PMID- 14764540 TI - Reconstitution of dendritic and natural killer-cell subsets after allogeneic stem cell transplantation: effects of endogenous flt3 ligand. AB - Recovery of dendritic cells (DCs) and natural killer (NK) cells after allogeneic stem cell transplantation (SCT) is important for allograft responses and antitumor immunity and thus for treatment outcome. Regulation of this regenerative process is not well understood. We investigated the influence of endogenous cytokines on the recovery and diversification of DC and NK cell subsets up to 6 months after SCT. Reconstitution of circulating DCs and NK cells was rapid but accompanied by prolonged skewing of cell subsets. The speed of recovery of CD11c(+)CD123(low) DC1 exceeded that of CD11c(-) CD123(+) DC2, and correlated with plasma levels of flt3 ligand (FL), but not with granulocyte or granulocyte-macrophage colony-stimulating factors and stem cell factor. There was a 5-fold increase in interferon-gamma-producing CD56(high)CD16(-)/low NK cells and a corresponding reduction in the CD56(low)CD16(high) subset, accompanied by strongly reduced NK cell cytotoxicity. In vitro data implicate an inhibitory effect of cyclosporin A on NK cell differentiation and cytotoxicity. NK cell numbers did not correlate with plasma levels of FL or interleukin 15. Our results demonstrate that endogenous FL has distinct effects on the kinetics of reconstitution of DCs and NK cells and have potential implications for the modulation of immune responses after allogeneic SCT. PMID- 14764542 TI - Preference of simple sequence repeats in coding and non-coding regions of Arabidopsis thaliana. AB - MOTIVATION: Simple sequence repeats or microsatellites have been found abundantly in many genomes. However, the significance of distribution preference has not been completely understood. Completion of the Arabidopsis genome sequencing allows us to better understand and characterize microsatellites. RESULTS: Microsatellite distribution was more abundant in 5'-flanking regions of genes compared with that expected in the whole genome, with an over-representation of AG and AAG repeats; there were clear differences from distributions in 3'-flanks and coding fractions, where triplet frequencies evidently corresponded to codon usage. We identified 1140 full-length genes that contained at least one locus of AG or AAG repeats in their upstream sequences, and whose functional characteristics were significantly associated with the repeats. This observation indicates that selective pressure markedly differed in the three transcribed regions, with positive selection of AG and AAG repeats in 5'-flanks close to those genes whose products are preferentially involved in transcription. PMID- 14764541 TI - Kostmann syndrome: severe congenital neutropenia associated with defective expression of Bcl-2, constitutive mitochondrial release of cytochrome c, and excessive apoptosis of myeloid progenitor cells. AB - Kostmann syndrome, or severe congenital neutropenia (SCN), is an autosomal recessive disorder of neutrophil production. To investigate the potential role of apoptosis in SCN, bone marrow aspirates and biopsies were obtained from 4 patients belonging to the kindred originally described by Kostmann and 1 patient with SCN of unknown inheritance. An elevated degree of apoptosis was observed in the bone marrow of these patients, and a selective decrease in B-cell lymphoma-2 (Bcl-2) expression was seen in myeloid progenitor cells. Furthermore, in vitro apoptosis of bone marrow-derived Kostmann progenitor cells was increased, and mitochondrial release of cytochrome c was detected in CD34(+) and CD33(+) progenitors from patients, but not in controls. Administration of granulocyte colony-stimulating factor (G-CSF) restored Bcl-2 expression and improved survival of myeloid progenitor cells. In addition, cytochrome c release was partially reversed upon incubation of progenitor cells with G-CSF. In sum, these studies establish a role for mitochondria-dependent apoptosis in the pathogenesis of Kostmann syndrome and yield a tentative explanation for the beneficial effect of growth factor administration in these patients. PMID- 14764543 TI - Large-scale assessment of the utility of low-resolution protein structures for biochemical function assignment. AB - MOTIVATION: Several protein function prediction methods employ structural features captured in three-dimensional (3D) descriptors of biologically relevant sites. These methods are successful when applied to high-resolution structures, but their detection ability in lower resolution predicted structures has only been tested for a few cases. RESULTS: A method that automatically generates a library of 3D functional descriptors for the structure-based prediction of enzyme active sites (automated functional templates, 593 in total for 162 different enzymes), based on functional and structural information automatically extracted from public databases, has been developed and evaluated using decoy structures. The applicability to predicted structures was investigated by analyzing decoys of varying quality, derived from enzyme native structures. For 35% of decoy structures, our method identifies the active site in models having 3-4 A coordinate root mean square deviation from the native structure, a quality that is reachable using state of the art protein structure prediction algorithms. AVAILABILITY: See http://www.bioinformatics.buffalo.edu/resources/aft/ PMID- 14764544 TI - SPI: a tool for incorporating gene expression data into a four-dimensional database of Caenorhabditis elegans embryogenesis. AB - MOTIVATION: A comprehensive gene expression database is essential for computer modeling and simulation of biological phenomena, including development. Development is a four-dimensional (4D; 3D structure and time course) phenomenon. We are constructing a 4D database of gene expression for the early embryogenesis of the nematode Caenorhabditis elegans. As a framework of the 4D database, we have constructed computer graphics (CG), into which we will incorporate the expression data of a number of genes at the subcellular level. However, the assignment of 3D distribution of gene products (protein, mRNA), of embryos at various developmental stages, is both difficult and tedious. We need to automate this process. For this purpose, we developed a new system, named SPI after superimposing fluorescent confocal microscopic data onto a CG framework. RESULTS: The scheme of this system comprises the following: (1) acquirement of serial sections (40 slices) of fluorescent confocal images of three colors (4',6' diamino-2-phenylindole (DAPI) for nuclei, indodicarbocyanine (Cy-3) for the internal marker, which is a germline-specific protein POS-1 and indocarbocyanine (Cy-5) for the gene product to be examined); (2) identification of several features of the stained embryos, such as contour, developmental stage and position of the internal marker; (3) selection of CG images of the corresponding stage for template matching; (4) superimposition of serial sections onto the CG; (5) assignment of the position of superimposed gene products. The Snakes algorithm identified the embryo contour. The detection accuracy of embryo contours was 92.1% when applied to 2- to 28-cell-stage embryos. The accuracy of the developmental stage prediction method was 81.2% for 2- to 8-cell-stage embryos. We manually judged only the later stage embryos because the accuracy for embryos at the later stages was unsatisfactory due to experimental noise effects. Finally, our system chose the optimal CG and performed the superposition and assignment of gene product distribution. We established an initial 4D gene expression database with 56 maternal gene products. AVAILABILITY: This system is available at http://anti.lab.nig.ac.jp/spi/ and http://anti.lab.nig.ac.jp/4ddb/ PMID- 14764545 TI - Exploring genome architecture through GOV: a WWW-based gene order visualizer. AB - MOTIVATION: The past decade has seen extension in the methods of sequence analysis from single gene based to analyzing multiple genes and proteins simultaneously. Consequently, there is a need for software tools that will allow mining of these enormous datasets at genome level effectively. A key challenge is to make them user-friendly, available to a larger community and integrate with public domain software without much hassle. RESULTS: A web-based interactive computational tool is described for visualization and comparison of gene order from prokaryotic and selected viral genome data. Many intriguing similarities and differences in gene order of multiple genomes can be compared and revealed. The interface facilitates easy extraction of the nucleotide sequence of the gene of interest and BLAST analysis against GenBank at NCBI to provide insights into gene functions and orthologs of the gene in other species. PMID- 14764546 TI - Confirmation of data mining based predictions of protein function. AB - MOTIVATION: A central problem in bioinformatics is the assignment of function to sequenced open reading frames (ORFs). The most common approach is based on inferred homology using a statistically based sequence similarity (SIM) method, e.g. PSI-BLAST. Alternative non-SIM based bioinformatic methods are becoming popular. One such method is Data Mining Prediction (DMP). This is based on combining evidence from amino-acid attributes, predicted structure and phylogenic patterns; and uses a combination of Inductive Logic Programming data mining, and decision trees to produce prediction rules for functional class. DMP predictions are more general than is possible using homology. In 2000/1, DMP was used to make public predictions of the function of 1309 Escherichia coli ORFs. Since then biological knowledge has advanced allowing us to test our predictions. RESULTS: We examined the updated (20.02.02) Riley group genome annotation, and examined the scientific literature for direct experimental derivations of ORF function. Both tests confirmed the DMP predictions. Accuracy varied between rules, and with the detail of prediction, but they were generally significantly better than random. For voting rules, accuracies of 75-100% were obtained. Twenty-one of these DMP predictions have been confirmed by direct experimentation. The DMP rules also have interesting biological explanations. DMP is, to the best of our knowledge, the first non-SIM based prediction method to have been tested directly on new data. AVAILABILITY: We have designed the "Genepredictions" database for protein functional predictions. This is intended to act as an open repository for predictions for any organism and can be accessed at http://www.genepredictions.org PMID- 14764547 TI - PDBSprotEC: a Web-accessible database linking PDB chains to EC numbers via SwissProt. AB - A mapping between chains in the Protein Databank and Enzyme Classification numbers is invaluable for research into structure-function relationships. Mapping at the chain level is a non-trivial problem and we present an automatically updated Web-server, which provides this link in a queryable form and as a downloadable XML or flat file. PMID- 14764548 TI - Predicting rules on organization of cis-regulatory elements, taking the order of elements into account. AB - MOTIVATION: In eukaryotes, rules regarding organization of cis-regulatory elements are complex. They sometimes govern multiple kinds of elements and positional restrictions on elements. RESULTS: We propose a method for detecting rules, by which the order of elements is restricted. The order restriction is expressed as element patterns. We extract all the element patterns that occur in promoter regions of at least the specified number of genes. Then, we find significant patterns based on the expression similarity of genes with promoter regions containing each of the extracted patterns. When we applied our method to Saccharomyces cerevisiae, we detected significant patterns overlooked by previous methods, thus demonstrating the utility of our method for analyses of eukaryotic gene regulation. We also suggest that several types of element organization exist: (i) those in which only the order of elements is important, (ii) order and distance both are important and (iii) only the combination of elements is important. AVAILABILITY: The program for extracting element patterns is available upon request. PMID- 14764549 TI - CompuCell, a multi-model framework for simulation of morphogenesis. AB - MOTIVATION: CompuCell is a multi-model software framework for simulation of the development of multicellular organisms known as morphogenesis. It models the interaction of the gene regulatory network with generic cellular mechanisms, such as cell adhesion, division, haptotaxis and chemotaxis. A combination of a state automaton with stochastic local rules and a set of differential equations, including subcellular ordinary differential equations and extracellular reaction diffusion partial differential equations, model gene regulation. This automaton in turn controls the differentiation of the cells, and cell-cell and cell extracellular matrix interactions that give rise to cell rearrangements and pattern formation, e.g. mesenchymal condensation. The cellular Potts model, a stochastic model that accurately reproduces cell movement and rearrangement, models cell dynamics. All these models couple in a controllable way, resulting in a powerful and flexible computational environment for morphogenesis, which allows for simultaneous incorporation of growth and spatial patterning. RESULTS: We use CompuCell to simulate the formation of the skeletal architecture in the avian limb bud. AVAILABILITY: Binaries and source code for Microsoft Windows, Linux and Solaris are available for download from http://sourceforge.net/projects/compucell/ PMID- 14764550 TI - AmpliBASE MT: a Mycobacterium tuberculosis diversity knowledgebase. AB - AmpliBASE MT is an online databank of high-resolution DNA fingerprints representing fluorescent amplified fragment length polymorphism (FAFLP) profiles or amplitypes developed for the Mycobacterium tuberculosis complex strains from 48 different countries. AmpliBASE MT is based on a relational database management system that is hyperlinked to visualize genotyping results in the form of DNA fingerprint images for individual strains. A flexible search system based on systematic comparisons of fragment sizes in base pairs allows inter-laboratory comparison of FAFLP profiles. Besides this, the database also displays previously published data on IS6110 profiles, spoligotypes, MIRU-VNTRs and large sequence polymorphisms along with the FAFLP records that will give the overall comparisons. Being the first of its kind, AmpliBASE MT is expected to be a very helpful tool in strengthening the concept of 'geographic genomics' and will be very helpful to molecular epidemiologists and those interested in diagnostic development for tuberculosis. PMID- 14764551 TI - ChromoViz: multimodal visualization of gene expression data onto chromosomes using scalable vector graphics. AB - SUMMARY: ChromoViz is an R package for the visualization of microarray gene expression data, cross-species and cross-platform comparisons, as well as non expression genomic data obtained from public databases onto chromosomes. Chromosomal visualization format is proposed for the clear decoupling of the data layer from the procedure layer and the combined visualization of genomic data from heterogeneous data sources. Visualization with Javascript-enabled scalable vector graphics enables interactive visualization and navigation of data objects on the Web. AVAILABILITY: http://www.snubi.org/software/ChromoViz/ PMID- 14764552 TI - Large-scale co-evolution analysis of protein structural interlogues using the global protein structural interactome map (PSIMAP). AB - MOTIVATION: Interacting pairs of proteins should co-evolve to maintain functional and structural complementarity. Consequently, such a pair of protein families shows similarity between their phylogenetic trees. Although the tendency of co evolution has been known for various ligand-receptor pairs, it has not been studied systematically in the widest possible scope. We investigated the degree of co-evolution for more than 900 family pairs in a global protein structural interactome map (PSIMAP--a map of all the structural domain-domain interactions in the PDB). RESULTS: There was significant correlation in 45% of the total SCOPs Family level pairs, rising to 78% in 454 reliable family interactions. Expectedly, the intra-molecular interactions between protein families showed stronger co-evolution than inter-molecular interactions. However, both types of interaction have a fundamentally similar pattern of co-evolution except for cases where different interfaces are involved. These results validate the use of co evolution analysis with predictive methods such as PSIMAP to improve the accuracy of prediction based on "homologous interaction". The tendency of co-evolution enabled a nearly 5-fold enrichment in the identification of true interactions among the potential interlogues in PSIMAP. The estimated sensitivity was 79.2%, and the specificity was 78.6%. AVAILABILITY: The results of co-evolution analysis are available online at http://www.biointeraction.org PMID- 14764554 TI - Multiple sequence alignment in parallel on a workstation cluster. AB - SUMMARY: Multiple sequence alignment is the NP-hard problem of aligning three or more DNA or amino acid sequences in an optimal way so as to match as many characters as possible from the set of sequences. The popular sequence alignment program ClustalW uses the classical method of approximating a sequence alignment, by first computing a distance matrix and then constructing a guide tree to show the evolutionary relationship of the sequences. We show that parallelizing the ClustalW algorithm can result in significant speedup. We used a cluster of workstations using C and message passing interface for our implementation. Experimental results show that speedup of over 5.5 on six processors is obtainable for most inputs. AVAILABILITY: The software is available upon request from the second author. PMID- 14764553 TI - Predicting subcellular localization of proteins in a hybridization space. AB - MOTIVATION: The localization of a protein in a cell is closely correlated with its biological function. With the number of sequences entering into databanks rapidly increasing, the importance of developing a powerful high-throughput tool to determine protein subcellular location has become self-evident. In view of this, the Nearest Neighbour Algorithm was developed for predicting the protein subcellular location using the strategy of hybridizing the information derived from the recent development in gene ontology with that from the functional domain composition as well as the pseudo amino acid composition. RESULTS: As a showcase, the same plant and non-plant protein datasets as investigated by the previous investigators were used for demonstration. The overall success rate of the jackknife test for the plant protein dataset was 86%, and that for the non-plant protein dataset 91.2%. These are the highest success rates achieved so far for the two datasets by following a rigorous cross-validation test procedure, suggesting that such a hybrid approach (particularly by incorporating the knowledge of gene ontology) may become a very useful high-throughput tool in the area of bioinformatics, proteomics, as well as molecular cell biology. AVAILABILITY: The software would be made available on sending a request to the authors. PMID- 14764555 TI - CLENCH: a program for calculating Cluster ENriCHment using the Gene Ontology. AB - SUMMARY: Analysis of microarray data most often produces lists of genes with similar expression patterns, which are then subdivided into functional categories for biological interpretation. Such functional categorization is most commonly accomplished using Gene Ontology (GO) categories. Although there are several programs that identify and analyze functional categories for human, mouse and yeast genes, none of them accept Arabidopsis thaliana data. In order to address this need for A.thaliana community, we have developed a program that retrieves GO annotations for A.thaliana genes and performs functional category analysis for lists of genes selected by the user. AVAILABILITY: http://www.personal.psu.edu/nhs109/Clench PMID- 14764556 TI - Cluster Analyzer for Transcription Sites (CATS): a C++-based program for identifying clustered transcription factor binding sites. AB - SUMMARY: We have developed a program, Cluster Analyzer for Transcription Sites (CATS), which identifies clusters of transcription factor binding sites in any genome sequence. The program searches for clusters of the consensus sequence for DNA binding within a window (length of DNA). The window size and the cluster size (number of consensus sequences within a given window) can be varied. CATS can be used for single or multiple transcription factors for which consensus sequences have been deduced based on biochemical and mutational analysis, or by comparative genomics. The use of CATS for clusters of different transcription factor binding sites may facilitate the identification of genes that are co-regulated in a cell type-specific or developmental stage-specific manner. CATS is simple to install and use on computers running any Windows NT-platforms. AVAILABILITY: http://www.healthsciences.columbia.edu/dept/greenwaldlab/links.html PMID- 14764557 TI - Gene structure prediction from consensus spliced alignment of multiple ESTs matching the same genomic locus. AB - MOTIVATION: Accurate gene structure annotation is a challenging computational problem in genomics. The best results are achieved with spliced alignment of full length cDNAs or multiple expressed sequence tags (ESTs) with sufficient overlap to cover the entire gene. For most species, cDNA and EST collections are far from comprehensive. We sought to overcome this bottleneck by exploring the possibility of using combined EST resources from fairly diverged species that still share a common gene space. Previous spliced alignment tools were found inadequate for this task because they rely on very high sequence similarity between the ESTs and the genomic DNA. RESULTS: We have developed a computer program, GeneSeqer, which is capable of aligning thousands of ESTs with a long genomic sequence in a reasonable amount of time. The algorithm is uniquely designed to tolerate a high percentage of mismatches and insertions or deletions in the EST relative to the genomic template. This feature allows use of non-cognate ESTs for gene structure prediction, including ESTs derived from duplicated genes and homologous genes from related species. The increased gene prediction sensitivity results in part from novel splice site prediction models that are also available as a stand-alone splice site prediction tool. We assessed GeneSeqer performance relative to a standard Arabidopsis thaliana gene set and demonstrate its utility for plant genome annotation. In particular, we propose that this method provides a timely tool for the annotation of the rice genome, using abundant ESTs from other cereals and plants. AVAILABILITY: The source code is available for download at http://bioinformatics.iastate.edu/bioinformatics2go/gs/download.html. Web servers for Arabidopsis and other plant species are accessible at http://www.plantgdb.org/cgi-bin/AtGeneSeqer.cgi and http://www.plantgdb.org/cgi bin/GeneSeqer.cgi, respectively. For non-plant species, use http://bioinformatics.iastate.edu/cgi-bin/gs.cgi. The splice site prediction tool (SplicePredictor) is distributed with the GeneSeqer code. A SplicePredictor web server is available at http://bioinformatics.iastate.edu/cgi-bin/sp.cgi PMID- 14764558 TI - An evaluation of new criteria for CpG islands in the human genome as gene markers. AB - MOTIVATION: Recently, more stringent criteria for CpG islands have been introduced to exclude Alu repeats, thereby enabling a higher proportion of CpG islands associating with genes to be identified. Using these new criteria, several types of associations between CpG islands and genes were investigated to further establish the importance of CpG islands as gene markers. RESULTS: The CpG islands were searched by CpGIE, a java software program developed for CpG island identification. CpGIE was advanced in identification accuracy compared with other tools. According to our results, about 70% of the identified CpG islands were associating with the human genes and over half of them are in the promoters. Furthermore, the investigation of genes in the confirmed gene model showed that 56% of them had a CpG island overlapping the transcription start sites. In comparison, the new criteria were found capable of filtering a large fraction of Alu repeats that was identified as CpG islands by the generally accepted criteria within the genes, but very few CpG islands associating with the promoters were affected. The genes in the predicted gene model were not obviously associated with CpG islands, suggesting that CpG islands can be used to evaluate the accuracy of gene annotation. AVAILABILITY: http://bioinfo.hku.hk/cpgieintro PMID- 14764559 TI - A rapid method for illustrating features in both coding and non-coding regions of a genome. AB - A bitmap display of the Fourier spectra has been developed which allows convenient whole chromosome scanning for genes and other features. Use of a limited sliding window gives rapid visualization and localization of coding regions in the chromosomes, as well as non-coding features such as repetitive DNA. The method works particularly well on organisms with a skewed base composition, to provide an overview of genomic features. PMID- 14764560 TI - Metrics for comparing regulatory sequences on the basis of pattern counts. AB - MOTIVATION: Upstream sequences contain short motifs, which mediate transcriptional regulation by specifically binding different transcription factors. The presence of common motifs in the regulatory regions of two genes might be considered as a clue for a potential co-regulation. A pattern count based (dis)similarity metric between sequences could thus be used to classify genes according to their putative regulatory properties. RESULTS: We present here several metrics which rely on probability theory, and which aim at comparing sequences on the basis of pattern counts. We compare these metrics to several classical dissimilarity and similarity metrics, and illustrate their behaviour with a biological example. PMID- 14764561 TI - Rapid and sensitive dot-matrix methods for genome analysis. AB - MOTIVATION: Dot-matrix plots are widely used for similarity analysis of biological sequences. Many algorithms and computer software tools have been developed for this purpose. Though some of these tools have been reported to handle sequences of a few 100 kb, analysis of genome sequences with a length of >10 Mb on a microcomputer is still impractical due to long execution time and computer memory requirement. RESULTS: Two dot-matrix comparison methods have been developed for analysis of large sequences. The methods initially locate similarity regions between two sequences using a fast word search algorithm, followed with an explicit comparison on these regions. Since the initial screening removes most of random matches, the computing time is substantially reduced. The methods produce high quality dot-matrix plots with low background noise. Space requirements are linear, so the algorithms can be used for comparison of genome size sequences. Computing speed may be affected by highly repetitive sequence structures of eukaryote genomes. A dot-matrix plot of Yeast genome (12 Mb) with both strands was generated in 80 s with a 1 GHz personal computer. PMID- 14764562 TI - CHOISS for selection of single nucleotide polymorphism markers on interval regularity. AB - We developed algorithms that find a set of single nucleotide polymorphism (SNP) markers based on interval regularity, given either the number of SNPs to choose (m) or the desired interval (I), subject to minimum variance or minimum sum of squared deviations from I. In both cases, the number of all possible sets increases exponentially with respect to the number of input SNPs (n), but our algorithms find the minima only with O(n(2)) calculations and comparisons by elimination of redundancy. PMID- 14764563 TI - Comparison of various algorithms for recognizing short coding sequences of human genes. AB - MOTIVATION: Since the early 1980s of the twentieth century, there has been great progress in the development of computational gene-finding algorithms. Some problems, however, have not yet been solved currently. Recognizing short genes in prokaryotes and short exons in eukaryotes is one of such problems. The paper is devoted to assessing various algorithms, including those currently available and the new ones proposed here, in order to find the best algorithm to solve the issue. RESULTS: The databases consisting of phase-specific coding and non-coding sequences of human genes with length of 192, 162, 129, 108, 87, 63 and 42 bp, respectively, have been established. Based on the databases and a standard benchmark, 19 algorithms were evaluated, which include the methods of Markov models with orders of 1 through 5, codon usage, hexamer usage, codon preference, amino acid usage, codon prototype, Fourier transform and 8 Z curve methods with various numbers of parameters. Consequently, the Z curve methods with 69 and 189 parameters are the best ones among them, based on the databases constructed here. In addition to the highest recognition accuracy confirmed by 10-fold cross validation tests, the Z curve methods are much simpler computationally than the second best one, the fifth-order Markov chain model, in which 12 288 parameters are used. We hope that the Z curve methods presented in this paper would be beneficial to the further development of gene-finding algorithms. AVAILABILITY: The programs of various Z curve methods are available on request. PMID- 14764564 TI - Adplot: detection and visualization of repetitive patterns in complete genomes. AB - MOTIVATION: Repetitive DNA sequences are abundant in genomes and efficient mining of significant repeats is important as the first step of repetitive sequence research. Although many computational tools for the purpose, either automatic or visualization ones, have been developed, detection and analysis of approximate repeats are still non-trivial task. RESULTS: Auto Dot PLOT (Adplot), a dotplot like repetitive pattern visualization program with a window filtering based on iid Bernoulli trials, is developed and applied to yeast chromosomes and human T cell receptor locus sequence. Typical examples found in yeast chromosomes 1 and 10 and a tandem repeat of periods longer than 10,000 bp in human T cell receptor locus are presented. A complex structure composed of both direct and palindromic repeats found in yeast chromosome 10 is also visualized as specific dot pattern. Computational time measured by a Pentium 3 PC for each yeast auto chromosome with a standard parameter setting is linearly scaled and below 10 s per one chromosome, indicating efficiency of the program. From the examples, it is shown that Adplot can visualize approximate local repeat structures and give us a diagnosis power for inferring a duplicational history of repeats. AVAILABILITY: Adplot can be obtained by an e-mail request. PMID- 14764565 TI - FORTE: a profile-profile comparison tool for protein fold recognition. AB - We present FORTE, a profile-profile comparison tool for protein fold recognition. Users can submit a protein sequence to explore the possibilities of structural similarity existing in known structures. Results are reported via email in the form of pairwise alignments. PMID- 14764566 TI - Plotting haplotype-specific linkage disequilibrium patterns by extended haplotype homozygosity. AB - Association studies may request more details of a specific haplotype. Haplotype specific decay of linkage disequilibrium is such a crucial and versatile characteristic. It may be used, e.g. to search for signals of natural selection in a risk haplotype. Here, we present a web-based tool to explore the relationship between population frequency and extended linkage disequilibrium measured as haplotype homozygosity of observed haplotypes within a specified candidate region. AVAILABILITY: The web-tool is available at http://ihg.gsf.de/cgi-bin/mueller/webehh.pl PMID- 14764567 TI - Fold-specific substitution matrices for protein classification. AB - MOTIVATION: Methods that focus on secondary structures, such as Position Specific Scoring Matrices and Hidden Markov Models, have proved useful for assigning proteins to families. However, for assigning proteins to an attribute class within a family these methods may introduce more free parameters than are needed. There are fewer members and there is less variability among sequences within a family. We describe a method for organizing proteins in a family that exhibits up to an order of magnitude reduction in the number of parameters. The basis is the log odds ratio commonly used to measure similarity. We adapt this to characterize the sequence dissimilarities that give rise to attribute differentiation. This leads to the definition of Class Attribute Substitution Matrices (CLASSUM), a dual of the BLOSUM. RESULTS: The method was applied to classify sequences hierarchically in the lambda and kappa subgroups of the immunoglobulin superfamily. Positions conferring class were identified based on the degree of amino acid variability at a position. The CLASSUM computed for these positions classified better than 90% of test data correctly compared with 35-50% for BLOSUM 62. The expected value for a random matrix is 14%. The results suggest that family-specific data-derived substitution matrices can improve the resolution of automated methods that use generic substitution matrices for searching for and classifying proteins. PMID- 14764568 TI - Sequence analysis and membrane partitioning energies of alpha-helical antimicrobial peptides. AB - Sequences of 221 alpha-helical antimicrobial peptides (alphaAMPs) were compared and 63-166 of them were selected and analyzed using Perl programs. The results showed that aliphatic amino acids Gly, Leu, Ala, Ile and two positively charged amino acids Lys and Arg were composed of more than 63% of the first 20 residues of alphaAMPs. The weighed mean membrane partitioning energies at positions from 1 to 25 of alphaAMPs were calculated. Profile of the partitioning energies suggests oblique membrane insertion and an amphipathic alpha-helical structure of the N terminus of alphaAMP (residues from 1 to 13), a bend structure at positions 13 and 14, and a less structured C-terminus that parallels the surface of the membrane. These structural features are in good agreement with the experimentally determined membrane structure of hemagglutinin fusion peptide from influenza virus. We hypothesize that this (N-terminal oblique alpha-helix)-central bend-(C terminus) could be a common structural motif of membrane-disruptive peptides. PMID- 14764569 TI - SQUARE--determining reliable regions in sequence alignments. AB - The Server for Quick Alignment Reliability Evaluation (SQUARE) is a Web-based version of the method we developed to predict regions of reliably aligned residues in sequence alignments. Given an alignment between a query sequence and a sequence of known structure, SQUARE is able to predict which residues are reliably aligned. The server accesses a database of profiles of sequences of known three-dimensional structures in order to calculate the scores for each residue in the alignment. SQUARE produces a graphical output of the residue profile-derived alignment scores along with an indication of the reliability of the alignment. In addition, the scores can be compared against template secondary structure, conserved residues and important sites. PMID- 14764570 TI - GelScape: a web-based server for interactively annotating, manipulating, comparing and archiving 1D and 2D gel images. AB - GelScape is a web-based tool that permits facile, interactive annotation, comparison, manipulation and storage of protein gel images. It uses Java applet servlet technology to allow rapid, remote image handling and image processing in a platform-independent manner. It supports many of the features found in commercial, stand-alone gel analysis software including spot annotation, spot integration, gel warping, image resizing, HTML image mapping, image overlaying as well as the storage of gel image and gel annotation data in compliance with Federated Gel Database requirements. PMID- 14764571 TI - Discrepancies in dbSNP confirmation rates and allele frequency distributions from varying genotyping error rates and patterns. AB - SUMMARY: Three recent publications have examined the quality and completeness of public database single nucleotide polymorphism (dbSNP) and have come to dramatically different conclusions regarding dbSNPs false positive rate and the proportion of dbSNPs that are expected to be common. These studies employed different genotyping technologies and different protocols in determining minimum acceptable genotyping quality thresholds. Because heterozygous sites typically have lower quality scores than homozygous sites, a higher minimum quality threshold reduces the number of false positive SNPs, but yields fewer heterozygotes and leads to fewer confirmed SNPs. To account for the different confirmation rates and distributions of minor allele frequencies, we propose that the three confirmation studies have different false positive and false negative rates. We developed a mathematical model to predict SNP confirmation rates and the apparent distribution of minor allele frequencies under user-specified false positive and false negative rates. We applied this model to the three published studies and to our own resequencing effort. We conclude that the dbSNP false positive rate is approximately 15-17% and that the reported confirmation studies have vastly different genotyping error rates and patterns. PMID- 14764572 TI - Genes@Work: an efficient algorithm for pattern discovery and multivariate feature selection in gene expression data. AB - MOTIVATION: Despite the growing literature devoted to finding differentially expressed genes in assays probing different tissues types, little attention has been paid to the combinatorial nature of feature selection inherent to large, high-dimensional gene expression datasets. New flexible data analysis approaches capable of searching relevant subgroups of genes and experiments are needed to understand multivariate associations of gene expression patterns with observed phenotypes. RESULTS: We present in detail a deterministic algorithm to discover patterns of multivariate gene associations in gene expression data. The patterns discovered are differential with respect to a control dataset. The algorithm is exhaustive and efficient, reporting all existent patterns that fit a given input parameter set while avoiding enumeration of the entire pattern space. The value of the pattern discovery approach is demonstrated by finding a set of genes that differentiate between two types of lymphoma. Moreover, these genes are found to behave consistently in an independent dataset produced in a different laboratory using different arrays, thus validating the genes selected using our algorithm. We show that the genes deemed significant in terms of their multivariate statistics will be missed using other methods. AVAILABILITY: Our set of pattern discovery algorithms including a user interface is distributed as a package called Genes@Work. This package is freely available to non-commercial users and can be downloaded from our website (http://www.research.ibm.com/FunGen). PMID- 14764573 TI - Good spaced seeds for homology search. AB - MOTIVATION: Filtration is an important technique used to speed up local alignment as exemplified in the BLAST programs. Recently, Ma et al. discovered that better filtering can be achieved by spacing out the matching positions according to a certain pattern, instead of contiguous positions to trigger a local alignment in their PatternHunter program. Such a match pattern is called a spaced seed. RESULTS: Our numerical computation shows that the ranks of spaced seeds (based on sensitivity) change with the sequences similarity. Since homologous sequences may have diverse similarity, we assess the sensitivity of spaced seeds over a range of similarity levels and present a list of good spaced seeds for facilitating homology search in DNA genomic sequences. We validate that the listed spaced seeds are indeed more sensitive using three arbitrarily chosen pairs of DNA genomic sequences. PMID- 14764574 TI - Sensitivity and specificity of five abundance estimators for high-density oligonucleotide microarrays. AB - MOTIVATION: A number of algorithms have been proposed for the processing of feature-level data from high-density oligonucleotide microarrays to give estimates of transcript abundance. Performance in the common task of detecting differential expression between samples can be quantified by the statistical concepts of sensitivity and specificity, and represented by the use of receiver operating characteristic curves. These have been previously presented for small numbers of genes known to be differentially present in spiked-in samples. We present here a study of performance over a large number (thousands) of transcripts for which there is strong evidence of differential expression, with corresponding false positive rates controlled by comparisons between replicates. RESULTS: The straight-line regression analysis of a mixture series with replicates by five estimation algorithms produces a consensus set of 4462 transcripts with differential expression of agreed direction and high significance (p < 0.01) according to all algorithms. The more difficult task of two-sample tests between adjacent mixture levels produces performance curves of fraction true positive detected against significance level. Performance varies significantly between algorithms: at the p < 0.01 level, the detection rate varies between 41 and 66%. A control using comparisons between replicates at the same levels indicates that the tests produce empirical false positive rates closely matching the nominal p-values. PMID- 14764575 TI - Statistically rigorous automated protein annotation. AB - MOTIVATION: Assignment of putative protein functional annotation by comparative analysis using pre-defined experimental annotations is performed routinely by molecular biologists. The number and statistical significance of these assignments remains a challenge in this era of high-throughput proteomics. A combined statistical method that enables robust, automated protein annotation by reliably expanding existing annotation sets is described. An existing clustering scheme, based on relevant experimental information (e.g. sequence identity, keywords or gene expression data) is required. The method assigns new proteins to these clusters with a measure of reliability. It can also provide human reviewers with a reliability score for both new and previously classified proteins. RESULTS: A dataset of 27 000 annotated Protein Data Bank (PDB) polypeptide chains (of 36 000 chains currently in the PDB) was generated from 23 000 chains classified a priori. AVAILABILITY: PDB annotations and sample software implementation are freely accessible on the Web at http://pmr.sdsc.edu/go PMID- 14764576 TI - Ontologizing gene-expression microarray data: characterizing clusters with Gene Ontology. AB - An XML-based Java application is described that provides a function-oriented overview of the results of cluster analysis of gene-expression microarray data based on Gene Ontology terms and associations. The application generates one HTML page with listings of the frequencies of explicit and implicit Gene Ontology annotations for each cluster, and separate, linked pages with listings of explicit annotations for each gene in a cluster. PMID- 14764577 TI - Recognition and analysis of protein-coding genes in severe acute respiratory syndrome associated coronavirus. AB - MOTIVATION: The recent outbreak of severe acute respiratory syndrome (SARS) caused by SARS coronavirus (SARS-CoV) has necessitated an in-depth molecular understanding of the virus to identify new drug targets. The availability of complete genome sequence of several strains of SARS virus provides the possibility of identification of protein-coding genes and defining their functions. Computational approach to identify protein-coding genes and their putative functions will help in designing experimental protocols. RESULTS: In this paper, a novel analysis of SARS genome using gene prediction method GeneDecipher developed in our laboratory has been presented. Each of the 18 newly sequenced SARS-CoV genomes has been analyzed using GeneDecipher. In addition to polyprotein 1ab(1), polyprotein 1a and the four genes coding for major structural proteins spike (S), small envelope (E), membrane (M) and nucleocapsid (N), six to eight additional proteins have been predicted depending upon the strain analyzed. Their lengths range between 61 and 274 amino acids. Our method also suggests that polyprotein 1ab, polyprotein 1a, S, M and N are proteins of viral origin and others are of prokaryotic. Putative functions of all predicted protein-coding genes have been suggested using conserved peptides present in their open reading frames. AVAILABILITY: Detailed results of GeneDecipher analysis of all the 18 strains of SARS-CoV genomes are available at http://www.igib.res.in/sarsanalysis.html PMID- 14764578 TI - Oxidant hypersensitivity of Fanconi anemia type C-deficient cells is dependent on a redox-regulated apoptotic pathway. AB - Fanconi anemia is a genetic disorder characterized by bone marrow failure. Significant evidence supports enhanced apoptosis of hematopoietic stem/progenitor cells as a critical factor in the pathogenesis of bone marrow failure in Fanconi anemia. However, the molecular mechanism(s) responsible for the apoptotic phenotype are incompletely understood. Here, we tested whether alterations in the activation of a redox-dependent pathway may participate in the pro-apoptotic phenotype of primary Fancc -/- cells in response to oxidative stress. Our data indicate that Fancc -/- cells are highly sensitive to oxidant stimuli and undergo enhanced oxidant-mediated apoptosis compared with wild type controls. In addition, antioxidants preferentially enhanced the survival of Fancc -/- cells. Because oxidative stress activates the redox-dependent ASK1 pathway, we assessed whether Fancc -/- cells exhibited increased oxidant-induced ASK1 activation. Our results revealed ASK1 hyperactivation in H2O2-treated Fancc -/- cells. Furthermore, using small interfering RNAs to decrease ASK1 expression and a dominant negative ASK1 mutant to inhibit ASK1 kinase activity, we determined that H2O2-induced apoptosis was ASK1-dependent. Collectively, these data argue that the predisposition of Fancc -/- hematopoietic stem/progenitor cells to apoptosis is mediated in part through altered redox regulation and ASK1 hyperactivation. PMID- 14764579 TI - Extracellular regulated kinase/mitogen activated protein kinase is up-regulated in pulmonary emphysema and mediates matrix metalloproteinase-1 induction by cigarette smoke. AB - The interstitial collagenase matrix metalloprotein-ase-1 (MMP-1) is up-regulated in the lung during pulmonary emphysema. The mechanisms underlying this aberrant expression are poorly understood. Although cigarette smoking is the predominant cause of emphysema, only 15-20% of smokers develop the disease. To define the signaling pathways activated by smoke and to identify molecules responsible for emphysema-associated MMP-1 expression, we performed several in vitro and in vivo experiments. In this study, we showed that cigarette smoke directly induced MMP-1 mRNA and protein expression and increased the collagenolytic activity of human airway cells. Treatment with various chemical kinase inhibitors revealed that this response was dependent on the extracellular regulated kinase-1/2 (ERK) mitogen activated protein kinase pathway. Cigarette smoke increased phosphorylation of residues Thr-202 and Tyr-204 of ERK in airway lining cells and alveolar macrophages in mice at 10 days and 6 months of exposure. Moreover, analysis of lung tissues from emphysema patients revealed significantly increased ERK activity compared with lungs of control subjects. This ERK activity was evident in airway lining and alveolar cells. The identification of active ERK in the lungs of emphysema patients and the finding that induction of MMP-1 by cigarette smoke in pulmonary epithelial cells is ERK-dependent reveal a molecular mechanism and potential therapeutic target for excessive matrix remodeling in smokers who develop emphysema. PMID- 14764580 TI - Mapping the interaction between high molecular mass kininogen and the urokinase plasminogen activator receptor. AB - The urokinase plasminogen activator receptor (uPAR) is a multifunctional, GPI linked receptor that modulates cell adhesion/migration and fibrinolysis. We mapped the interaction sites between soluble uPAR (suPAR) and high molecular mass kininogen (HK). Binding of biotin-HK to suPAR was inhibited by HK, 56HKa, and 46HKa with an IC50 of 60, 110, and 8 nm, respectively. We identified two suPAR binding sites, a higher affinity site in the light chain of HK and 46HKa (His477 Gly496) and a lower affinity site within the heavy chain (Cys333-Lys345). HK predominantly bound to suPAR fragments containing domains 2 and 3 (S-D2D3). Binding of HK to domain 1 (S-D1) was also detected, and the addition of S-D1 to S D2D3 completely inhibited biotin-HK or -46HKa binding to suPAR. Using sequential and overlapping 20-amino acid peptides prepared from suPAR, two regions for HK binding were identified. One on the carboxyl-terminal end of D2 (Leu166-Thr195) blocked HK binding to suPAR and to human umbilical vein endothelial cells (HUVEC). This site overlapped with the urokinase-binding region, and urokinase inhibited the binding of HK to suPAR. A second region on the amino-terminal portion of D3 (Gln215-Asn255) also blocked HK binding to HUVEC. Peptides that blocked HK binding to uPAR also inhibited prekallikrein activation on HUVEC. Therefore, HK interacts with suPAR at several sites. HK binds to uPAR as part of its interaction with its multiprotein receptor complex on HUVEC, and the biological functions that depend upon this binding are modulated by urokinase. PMID- 14764581 TI - Substrate and dioxygen binding to the endospore coat laccase from Bacillus subtilis. AB - The CotA laccase from the endospore coat of Bacillus subtilis has been crystallized in the presence of the non-catalytic co-oxidant 2,2'-azinobis-(3 ethylbenzothiazoline-6-sulfonate) (ABTS), and the structure was determined using synchrotron radiation. The binding site for this adduct is well defined and indicates how ABTS, in conjunction with laccases, could act as an oxidative mediator toward non-phenolic moieties. In addition, a dioxygen moiety is clearly defined within the solvent channel oriented toward one of the T3 copper atoms in the trinuclear center. PMID- 14764582 TI - The acetylcholinesterase homology region is essential for normal conformational maturation and secretion of thyroglobulin. AB - Secretion of thyroglobulin (Tg, a large homodimeric glycoprotein) is essential to deliver Tg to its site of iodination for thyroxine biosynthesis. An L2263P missense mutation in Tg has been proposed as the molecular defect causing congenital goitrous hypothyroidism in cog/cog mice due to perturbed Tg homodimerization, resulting in its retention within the endoplasmic reticulum. The mutation falls within a carboxyl-terminal region of Tg with high structural similarity to the entirety of acetylcholinesterase (AChE), a secretory protein that also forms homodimers. We provide new evidence that authentic AChE and the cholinesterase-like domain of Tg share a common tertiary structure. Moreover, we find that a Tg truncation, deleted of the cholinesterase-like region (but not a comparably sized deletion of internal Tg regions), blocks Tg export. Appending to this truncation a cDNA encoding authentic AChE results in translation of a chimeric protein in which AChE is present in a native, enzymatically active (albeit latent) conformation, and this fully rescues Tg secretion. Introduction of the cog mutation inhibits AChE enzyme activity, and established denaturing mutations of AChE block secretion of the Tg. Additional studies show that the native structure of the AChE region functions as a "dimerization domain," facilitating intracellular transport of Tg to the site of thyroid hormonogenesis. PMID- 14764583 TI - G Protein-coupled receptor kinase 2 regulator of G protein signaling homology domain binds to both metabotropic glutamate receptor 1a and Galphaq to attenuate signaling. AB - Heterotrimeric guanine nucleotide-binding (G) protein-coupled receptor kinases (GRKs) are cytosolic proteins that contribute to the adaptation of G protein coupled receptor signaling. The canonical model for GRK-dependent receptor desensitization involves GRK-mediated receptor phosphorylation to promote the binding of arrestin proteins that sterically block receptor coupling to G proteins. However, GRK-mediated desensitization, in the absence of phosphorylation and arrestin binding, has been reported for metabotropic glutamate receptor 1 (mGluR1) and gamma-aminobutyric acid B receptors. Here we show that GRK2 mutants impaired in Galphaq/11 binding (R106A, D110A, and M114A), bind effectively to mGluR1a, but do not mediate mGluR1a adaptation. Galphaq/11 is immunoprecipitated as a complex with mGluR1a in the absence of agonist, and either agonist treatment or GRK2 overexpression promotes the dissociation of the receptor/Galphaq/11 complex. However, these mGluR1a/Galphaq/11 interactions are not antagonized by the overexpression of either GRK2 mutants defective in Galphaq/11 binding or RGS4. We have also identified a GRK2-D527A mutant that binds Galphaq/11 in an AlF4(-)-dependent manner but is unable to either bind mGluR1a or attenuate mGluR1a signaling. We conclude that the mechanism underlying GRK2 phosphorylation-independent attenuation of mGluR1a signaling is RH domain dependent, requiring the binding of GRK2 to both Galphaq/11 and mGluR1a. This serves to coordinate GRK2 interactions with Galphaq/11 and to disrupt receptor/Galphaq/11 complexes. Our findings indicate that GRK2 regulates receptor/G protein interactions, in addition to its traditional role as a receptor kinase. PMID- 14764584 TI - T7 lysozyme represses T7 RNA polymerase transcription by destabilizing the open complex during initiation. AB - Bacteriophage T7 lysozyme binds to T7 RNA polymerase and inhibits transcription initiation and the transition from initiation to elongation. We have investigated each step of transcription initiation to determine where T7 lysozyme has the most effect. Stopped flow and equilibrium DNA binding studies indicate that T7 lysozyme does not inhibit the formation of the preinitiation open complex (open complex in the absence of initiating nucleotide). T7 lysozyme, however, does prevent the formation of a fully open initiation complex (open complex in the presence of the initiating nucleotide). This is consistent with the results that in the presence of T7 lysozyme the rate of G ladder RNA synthesis is about 5-fold slower and the GTP Kd is about 2-fold higher, but T7 lysozyme does not inhibit the initial rate of RNA synthesis with a premelted bulge-6 promoter (bubble from 4 to +2). Neither the RNA synthesis rate nor the extent of promoter opening is restored by increasing the initiating nucleotide concentration, indicating that T7 lysozyme represses transcription by interfering with the formation of a stable and a fully open initiation bubble or by altering the structure of the DNA in the initiation complex. As a consequence of the unstable initiation bubble and/or the inhibition of the conformational changes in the N-terminal domain of T7 RNAP, T7 lysozyme causes an increased production of abortive products from 2- to 5-mer that delays the transition from the initiation to the elongation phase. PMID- 14764585 TI - Vav1 transduces T cell receptor signals to the activation of the Ras/ERK pathway via LAT, Sos, and RasGRP1. AB - Vav1 is a signaling protein required for both positive and negative selection of CD4(+)CD8(+) double positive thymocytes. Activation of the ERK MAPK pathway is also required for positive selection. Previous work has shown that Vav1 transduces T cell receptor (TCR) signals leading to an intracellular calcium flux. We now show that in double positive thymocytes Vav1 is required for TCR induced activation of the ERK1 and ERK2 kinases via a pathway involving the Ras GTPase, and B-Raf, MEK1, and MEK2 kinases. Furthermore, we show that Vav1 transduces TCR signals to Ras by controlling the membrane recruitment of two guanine nucleotide exchange factors. First, Vav1 transduces signals via phospholipase Cgamma1 leading to the membrane recruitment of RasGRP1. Second, Vav1 is required for recruitment of Sos1 and -2 to the transmembrane adapter protein LAT. Finally, we show that Vav1 is required for TCR-induced LAT phosphorylation, a key event for the activation of both phospholipase Cgamma1 and Sos1/2. We propose that reduced LAT phosphorylation is the key reason for defective TCR-induced calcium flux and ERK activation in Vav1-deficient cells. PMID- 14764586 TI - Global suppression of IL-6-induced acute phase response gene expression after chronic in vivo treatment with the peroxisome proliferator-activated receptor alpha activator fenofibrate. AB - The peroxisome proliferator-activated receptor alpha (PPARalpha), which is highly expressed in liver, plays key roles in lipid metabolism and inflammation. Interleukin-6 (IL-6) is the principal inducer of acute phase response (APR) gene expression. In the present study, we demonstrate that chronic treatment with the PPARalpha agonist fenofibrate fully prevents the IL-6-induced APR gene expression in wild-type but not in PPARalpha-deficient mice. PPARalpha prevents the IL-6 induced expression of the positive APR genes fibrinogen-alpha, -beta, -gamma, haptoglobulin, and serum amyloid A and the IL-6-induced suppression of the negative APR gene, major urinary protein. Furthermore, the effect of PPARalpha on the APR gene expression does not simply consist in a delayed systemic response to IL-6 but occurs directly at the transcriptional level. This global suppression of acute phase gene transcription may be explained by two PPARalpha-dependent in vivo effects: 1) PPARalpha activation results in the down-regulation of the IL-6 receptor components gp80 and gp130 in the liver, thereby reducing the phosphorylation and activation of the downstream transcription factors STAT3 and c-Jun that transduce the IL-6 signal; and 2) PPARalpha reduces the basal expression of the transcription factors CCAAT enhancer-binding protein-alpha, beta, -delta, which are responsible for immediate and maintained transcription of APR genes. A similar global effect of fenofibrate on acute phase protein expression is observed in hyperlipidemic patients chronically treated with fenofibrate, which displayed decreased plasma concentrations of the positive APR proteins fibrinogen, C-reactive protein, serum amyloid A, plasminogen, and alpha2 macroglobulin and increased plasma concentrations of the negative APR albumin, underlining the clinical significance of our findings. PMID- 14764587 TI - Competition for access to the rat major histocompatibility complex class I peptide-loading complex reveals optimization of peptide cargo in the absence of transporter associated with antigen processing (TAP) association. AB - Major histocompatibility complex (MHC) class I molecules load peptides in the endoplasmic reticulum in a process during which the peptide cargo is normally optimized in favor of stable MHC-peptide interactions. A dynamic multimolecular assembly termed the peptide-loading complex (PLC) participates in this process and is composed of MHC class I molecules, calreticulin, ERp57, and tapasin bound to the transporter associated with antigen processing (TAP) peptide transporter. We have exploited the observation that the rat MHC class I allele RT1-Aa, when expressed in the rat C58 thymoma cell line, effectively competes and prevents the endogenous RT1-Au molecule from associating with TAP. However, stable RT1-Au molecules are assembled efficiently in competition with RT1-Aa, demonstrating that cargo optimization can occur in the absence of TAP association. Defined mutants of RT1-Aa, which do not allow formation of the PLC, fail to become thermostable in C58 cells. Wild-type RT1-Aa, which does allow PLC formation, also fails to become thermostable in this cell line, which carries the rat TAPB transporter that supplies peptides incompatible for RT1-Aa binding. Full optimization of RT1-Aa requires the presence of the TAP2A allele, which is capable of supplying suitable peptides. Thus, formation of the PLC alone is not sufficient for optimization of the MHC class I peptide cargo. PMID- 14764588 TI - A concerted mechanism for the suppression of a folding defect through interactions with chaperones. AB - Specific amino acid substitutions confer a temperature-sensitive-folding (tsf) phenotype to bacteriophage P22 coat protein. Additional amino acid substitutions, called suppressor substitutions (su), relieve the tsf phenotype. These su substitutions are proposed to increase the efficiency of procapsid assembly, favoring correct folding over improper aggregation. Our recent studies indicate that the molecular chaperones GroEL/ES are more effectively recruited in vivo for the folding of tsf:su coat proteins than their tsf parents. Here, the tsf:su coat proteins are studied with in vitro equilibrium and kinetic techniques to establish a molecular basis for suppression. The tsf:su coat proteins were monomeric, as determined by velocity sedimentation analytical ultracentrifugation. The stability of the tsf:su coat proteins was ascertained by equilibrium urea titrations, which were best described by a three-state folding model, N <--> I <--> U. The tsf:su coat proteins either had stabilized native or intermediate states as compared with their tsf coat protein parents. The kinetics of the I <--> U transition showed a decrease in the rate of unfolding and a small increase in the rate of refolding, thereby increasing the population of the intermediate state. The increased intermediate population may be the reason the tsf:su coat proteins are aggregation-prone and likely enhances GroEL-ES interactions. The N --> I unfolding rate was slower for the tsf:su proteins than their tsf coat parents, resulting in an increase in the native state population, which may allow more competent interactions with scaffolding protein, an assembly chaperone. Thus, the suppressor substitution likely improves folding in vivo through increased efficiency of coat protein-chaperone interactions. PMID- 14764589 TI - Disease-associated mutations in human mannose-binding lectin compromise oligomerization and activity of the final protein. AB - Deficiency of human mannose-binding lectin (MBL) caused by mutations in the coding part of the MBL2 gene is associated with increased risk and severity of infections and autoimmunity. To study the biological consequences of MBL mutations, we expressed wild type MBL and mutated MBL in Chinese hamster ovary cells. The normal MBL cDNA (WT MBL-A) was cloned, and the three known natural and two artificial variants were expressed in Chinese hamster ovary cells. When analyzed, WT MBL-A formed covalently linked higher oligomers with a molecular mass of about 300-450 kDa, corresponding to 12-18 single chains or 4-6 structural units. By contrast, all MBL variants formed a dominant band of about 50 kDa, with increasingly weaker bands at 75, 100, and 125 kDa corresponding to two, three, four, and five chains, respectively. In contrast to WT MBL-A, variant MBL formed noncovalent oligomers containing up to six chains (two structural units). MBL variants bound ligands with a markedly reduced capacity compared with WT MBL-A. Mutations in the collagenous region of human MBL compromise assembly of higher order oligomers, resulting in reduced ligand binding capacity and thus reduced capability to activate complement. PMID- 14764590 TI - Identification of a factor Xa-interactive site within residues 337-372 of the factor VIII heavy chain. AB - We recently demonstrated that the residues 337-372, comprising the acidic C terminal region in A1 subunit, interact with factor Xa during the proteolytic inactivation of factor VIIIa (Nogami, K., Wakabayashi, H., and Fay, P. J. (2003) J. Biol. Chem. 278, 16502-16509). We now show this sequence is important for factor Xa-catalyzed activation of factor VIII. Peptide 337-372 markedly inhibited cofactor activation, consistent with a delay in the rate of cleavage at the A1-A2 junction. Studies using the isolated factor VIII heavy chain indicated that the peptide completely blocked cleavage at the A1-A2 junction (IC50 = 11 microm) and partially blocked cleavage at the A2-B junction (IC50 = 100 microm). Covalent cross-linking was observed between the 337-372 peptide and factor Xa following reaction with 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide, and the peptide quenched the fluorescence of dansyl-Glu-Gly-Arg active site-modified factor Xa, suggesting that residues 337-372 directly interact with factor Xa. Studies using a monoclonal antibody recognizing residues 351-365 as well as the peptide to this sequence further restricted the interactive region. Mutant factor VIII molecules in which clustered acidic residues in the 337-372 segment were converted to alanine were evaluated for activation by factor Xa. Of the mutants tested, only factor Xa-catalyzed activation of the D361A/D362A/D363A mutant was inhibited with peak activity of approximately 50% and an activation rate constant of approximately 30% of the wild type values. These results indicate that the 337 372 acidic region separating A1 and A2 domains and, in particular, a cluster of acidic residues at position 361-363 contribute to a unique factor Xa-interactive site within the factor VIII heavy chain that promotes factor Xa docking during cofactor activation. PMID- 14764591 TI - Crystal structure of norwalk virus polymerase reveals the carboxyl terminus in the active site cleft. AB - Norwalk virus is a major cause of acute gastroenteritis for which effective treatments are sorely lacking. To provide a basis for the rational design of novel antiviral agents, the main replication enzyme in Norwalk virus, the virally encoded RNA-dependent RNA polymerase (RdRP), has been expressed in an enzymatically active form, and its structure has been crystallographically determined both in the presence and absence of divalent metal cations. Although the overall fold of the enzyme is similar to that seen previously in the RdRP from rabbit hemorrhagic disease virus, the carboxyl terminus, surprisingly, is located in the active site cleft in five independent copies of the protein in three distinct crystal forms. The location of this carboxyl-terminal segment appears to interfere with the binding of double-stranded RNA in the active site cleft and may play a role in the initiation of RNA synthesis or mediate interactions with accessory replication proteins. PMID- 14764592 TI - Patent ductus venosus and dioxin resistance in mice harboring a hypomorphic Arnt allele. AB - The Ah receptor nuclear translocator (ARNT) is the dimeric partner of hypoxia inducible factors and thus plays a pivotal role in cellular adaptation to low oxygen environments. ARNT is also a dimeric partner for the Ah receptor (AHR), and this complex is essential in regulating the adaptive metabolic response to polycyclic aromatic hydrocarbons. Because of the essential role of ARNT in hypoxia-driven developmental events, it has been difficult to study the physiological significance of AHR.ARNT heterodimers in vivo. To address this issue, we developed a hypomorphic Arnt allele that displayed normal development and allowed the examination of the role of ARNT in AHR biology. In this regard, the AHR is also known to mediate two additional biological processes: the toxicological response to compounds such as 2,3,7,8-tetrachlorodibenzo-p-dioxin (dioxin) and the developmental closure of a fetal vascular structure known as the ductus venosus. Although the mechanism of the adaptive pathway has been well described, the mechanism of AHR-mediated signal transduction in the toxic and developmental pathways is not well understood. Liver perfusion studies demonstrated that ARNT hypomorphs have a patent ductus venosus, identical to that observed in the Ahr null mice. Parallel dioxin toxicity studies demonstrated that the ARNT hypomorphs exhibited resistance to the end points of dioxin exposure. Moreover, we observed that toxicity could be segregated from the classical adaptive responses such as P4501A induction. Taken in sum, these experiments demonstrate that ARNT is an essential component of AHR developmental signaling and shed light on the mechanism of dioxin toxicity. PMID- 14764593 TI - Aryl hydrocarbon nuclear translocator (ARNT) promotes oxygen-independent stabilization of hypoxia-inducible factor-1alpha by modulating an Hsp90-dependent regulatory pathway. AB - Hypoxia-inducible factor-1 (HIF-1) is a potent cellular survival factor contributing to tumorigenesis in a broad range of cancers. The functional transcription factor exists as a heterodimeric complex consisting of HIF-1alpha and the aryl hydrocarbon receptor nuclear translocator (ARNT). Association of HIF 1 with ARNT is required for its activity; however, no other role has been ascribed to this interaction. We demonstrated previously that pharmacologic inhibition of Hsp90 by geldanamycin (GA) impairs HIF transcription and promotes VHL (Von Hippel-Lindau)-independent degradation of the protein, thus implicating Hsp90 as an essential interacting partner for HIF. In this study, we further explore the physiological role for Hsp90 in HIF function. We establish that the PAS (Per-ARNT-Sim) domain of HIF is required both to promote association with Hsp90 and confer sensitivity to GA. Coincidentally, this domain also associates with ARNT. Overexpression of ARNT in a VHL-deficient background resulted in substantially increased HIF-1 protein concomitant with increased protein stability. Conversely, down-regulation of endogenous ARNT protein by RNA interference decreased the steady-state HIF protein. ARNT-mediated stabilization of HIF is specific for the Hsp90-dependent pathway, as ARNT was unable to protect HIF from VHL-mediated degradation. We establish that the ability of ARNT to up regulate HIF and diminish HIF sensitivity to GA is due to its ability to compete for the Hsp90 binding site on HIF. These data elucidate novel functions for ARNT and Hsp90 in regulating HIF function and further illustrate that cofactor association may significantly impact upon the sensitivity of Hsp90 clients to chaperone inhibitors. PMID- 14764594 TI - Bcl-xL and E1B-19K proteins inhibit p53-induced irreversible growth arrest and senescence by preventing reactive oxygen species-dependent p38 activation. AB - In this study, we describe novel functions of the anti-apoptotic Bcl-2 family proteins. Bcl-x(L) and E1B-19K were found to inhibit p53-induced irreversible growth arrest and senescence, but not to inhibit transient growth arrest, implying that Bcl-x(L) and E1B-19K are specifically involved in senescence without participating in growth arrest. We provide several lines of evidences showing that the functions of Bcl-x(L) and E1B-19K to prevent generation of reactive oxygen species (ROS) are important to inhibit senescence induction. First, we found that that ROS are increased during p53-induced senescence. Moreover, Bcl-x(L) and E1B-19K inhibit this p53-induced ROS generation. Second, antioxidants prevent the induction of senescence and ROS by p53, but not the persistence of the senescence phenotype. Third, the anti-senescence functions of Bcl-x(L) and E1B-19K were suppressed by adding exogenous ROS. These results suggest that Bcl-x(L) and E1B-19K inhibit senescence induction by preventing ROS generation. Furthermore, p38 kinase was found to be activated during p53-induced senescence, but not in cells expressing Bcl-x(L) or E1B-19K, or in cells treated with anti-oxidants. Consistently, a chemical inhibitor of p38 kinase, SB203580, was found to inhibit p53-induced senescence, but only when treated before the cellular commitment to senescence, implying that p38 kinase is necessary for senescence induction. Therefore, Bcl-x(L) and E1B-19K inhibit p53-induced senescence by preventing ROS generation, which in turn leads to the activation of p38 kinase. These results also suggest that the oncogenic potential of Bcl-2 is due to its ability to inhibit senescence as well as apoptosis. PMID- 14764595 TI - An extracellular protein microdomain controls up-regulation of neuronal nicotinic acetylcholine receptors by nicotine. AB - In smoker's brain, rodent brain, and in cultured cells expressing nicotinic receptors, chronic nicotine treatment induces an increase in the total number of high affinity receptors for acetylcholine and nicotine, a process referred to as up-regulation. Up-regulation induced by 1 mm nicotine reaches 6-fold for alpha3beta2 nicotinic receptors transiently expressed in HEK 293 cells, whereas it is much smaller for alpha3beta4 receptors, offering a rationale to investigate the molecular mechanism underlying up-regulation. In this expression system binding sites are mainly intracellular, as shown by [(3)H]epibatidine binding experiments and competition with the impermeant ligand carbamylcholine. Systematic analysis of beta2/beta4 chimeras demonstrates the following. (i) The extracellular domain critically contributes to up-regulation. (ii) Only residues belonging to two beta2 segments, 74-89 and 106-115, confer up-regulation to beta4, mainly by decreasing the amount of binding sites in the absence of nicotine; on an atomic three-dimensional model of the alpha3beta2 receptor these amino acids form a compact microdomain that mainly contributes to the subunit interface and also faces the acetylcholine binding site. (iii) The beta4 microdomain is sufficient to confer to beta2 a beta4-like up-regulation. (iv) This microdomain makes an equivalent contribution to the up-regulation differences between alpha4beta2 and alpha4beta4. We propose that nicotine, by binding to immature oligomers, elicits a conformational reorganization of the microdomain, strengthening the interaction between adjacent subunits and, thus, facilitating maturation processes toward high affinity receptors. This mechanism may be central to nicotine addiction, since alpha4beta2 is the subtype exhibiting the highest degree of up-regulation in the brain. PMID- 14764596 TI - Control of the vascular endothelial growth factor internal ribosome entry site (IRES) activity and translation initiation by alternatively spliced coding sequences. AB - The vascular endothelial growth factor-A (VEGF) gene locus contains eight exons that span 14 kb. Alternative splicing generates multiple, different mRNAs that in turn translate into at least five protein isoforms. While the canonical AUG start codon is located at position 1039 in exon 1, there also exists an upstream, in frame CUG initiation codon that drives expression of L-VEGF, containing an additional 180 amino acids. Two separate internal ribosome entry sites (IRES) regulate the activity of each initiation codon. Thus the 5'-UTR of VEGF, which comprises the majority of exon 1, consists of IRES B, the CUG, IRES A, and the AUG, from 5' to 3'. Previously, it has been shown that IRES B regulates initiation at the CUG and IRES A regulates AUG usage. In this study, we have found evidence that the exon content of the VEGF mRNA, determined through alternative splicing, controls IRES A activity. While the CUG is most efficient at initiating translation, transcripts that lack both exons 6 and 7 and therefore contain an exon 5/8 junction lack AUG-initiated translation. The process of splicing is not responsible for this start codon selection since transfection of genomic and cDNA VEGF sequences give the same expression pattern. We hypothesize that long range tertiary interactions in the VEGF mRNA regulate IRES activity and thus control start codon selection. This is the first report describing the influence of alternatively spliced coding sequences on codon selection by modulating IRES activity. PMID- 14764597 TI - Peroxisome proliferator-activated receptor gamma activation modulates cyclin D1 transcription via beta-catenin-independent and cAMP-response element-binding protein-dependent pathways in mouse hepatocytes. AB - Activation of peroxisome proliferator-activated receptor gamma (PPARgamma) following exposure to PPARgamma-specific ligands resulted in growth inhibition in various carcinoma cell lines. Our aim was to elucidate the pathway of PPARgamma2 activation-mediated modulation of cyclin D1 transcription in mouse hepatocytes. To address this we utilized stable control and PPARgamma hepatocyte cell lines created via retroviral overexpression utilizing AML-12 hepatocytes. Addition of PPARgamma ligand troglitazone (TZD) activated PPARgamma2 in proliferating hepatocytes and resulted in growth arrest accompanied by a down-regulation of proliferating cell nuclear antigen, cyclin D1, and beta-catenin expression. Furthermore activation of PPARgamma2 attenuated cyclin D1 promoter activity indicating a transcriptional regulation of cyclin D1. Since beta-catenin plays a pivotal role in regulating cyclin D1 transcription, we studied whether PPARgamma2 mediated inhibition of cyclin D1 transcription involved beta-catenin. Interestingly overexpression of either wild-type or S37A mutant beta-catenin was unable to rescue PPARgamma2-mediated suppression of cyclin D1 transcription, whereas overexpression of cAMP-response element-binding protein (CREB) was capable of antagonizing this inhibitory effect of PPARgamma2. Additionally pretreatment with okadaic acid antagonized PPARgamma2-mediated inhibition of cyclin D1 transcription without any effect on beta-catenin expression. These studies also showed a TZD-mediated inhibition of total and phospho-CREB(Ser133) levels, CREB promoter activity, and cAMP-response element-mediated transcription in PPARgamma hepatocytes. Pretreatment of PPARgamma hepatocytes with okadaic acid, however, maintained higher total and phospho-CREB(Ser133) levels in the presence of TZD. These results indicated that PPARgamma2 activation inhibited cyclin D1 transcription in hepatocytes via CREB-dependent and beta-catenin independent pathways. PMID- 14764598 TI - Solution structure of the SEA domain from the murine homologue of ovarian cancer antigen CA125 (MUC16). AB - Human CA125, encoded by the MUC16 gene, is an ovarian cancer antigen widely used for a serum assay. Its extracellular region consists of tandem repeats of SEA domains. In this study we determined the three-dimensional structure of the SEA domain from the murine MUC16 homologue using multidimensional NMR spectroscopy. The domain forms a unique alpha/beta sandwich fold composed of two alpha helices and four antiparallel beta strands and has a characteristic turn named the TY turn between alpha1 and alpha2. The internal mobility of the main chain is low throughout the domain. The residues that form the hydrophobic core and the TY turn are fully conserved in all SEA domain sequences, indicating that the fold is common in the family. Interestingly, no other residues are conserved throughout the family. Thus, the sequence alignment of the SEA domain family was refined on the basis of the three-dimensional structure, which allowed us to classify the SEA domains into several subfamilies. The residues on the surface differ between these subfamilies, suggesting that each subfamily has a different function. In the MUC16 SEA domains, the conserved surface residues, Asn-10, Thr-12, Arg-63, Asp-75, Asp-112, Ser-115, and Phe-117, are clustered on the beta sheet surface, which may be functionally important. The putative epitope (residues 58-77) for anti-MUC16 antibodies is located around the beta2 and beta3 strands. On the other hand the tissue tumor marker MUC1 has a SEA domain belonging to another subfamily, and its GSVVV motif for proteolytic cleavage is located in the short loop connecting beta2 and beta3. PMID- 14764599 TI - Hyaluronan fragments stimulate endothelial recognition of injury through TLR4. AB - Tissues must quickly recognize injury to respond to the rapid pace of microbial growth. In skin, dermal microvascular endothelial cells must also react to danger signals from the surrounding tissue and immediately participate by initiating the wound repair process. Components of the extracellular matrix such as hyaluronan are rapidly broken down into smaller molecular weight oligosaccharides in a wound, and these can activate a variety of biological processes. This study set out to determine if hyaluronan fragments released following injury can stimulate endothelial cells and what mechanism is responsible for this response. Using genechip microarray analysis, a response to hyaluronan fragments was detected in endothelial cells with the most significant increase observed for the chemokine IL-8. This observation was verified with qualitative reverse transcriptase-PCR and ELISA in human endothelial cell culture, and in a mouse model by observing serum levels of MIP-2 and KC following hyaluronan fragment administration in vivo. Activation was TLR4-dependent, as shown by use of TLR4 blocking antibody and TLR4-deficient mice, but not due to the presence of undetected contaminants as shown by inactivation following digestion with the hyaluronan-degrading enzyme chondroitinase ABC or incubation with the hyaluronan-specific blocking peptide Pep-1. Inactivation of LPS activity failed to diminish the action of hyaluronan fragments. These observations suggest that endogenous components of the extracellular matrix can stimulate endothelia to trigger recognition of injury in the initial stages of the wound defense and repair response. PMID- 14764600 TI - Subunits of a yeast oligomeric G protein-coupled receptor are activated independently by agonist but function in concert to activate G protein heterotrimers. AB - G protein-coupled receptors (GPCRs) form dimeric or oligomeric complexes in vivo. However, the function of oligomerization in receptor-mediated G protein activation is unclear. Previous studies of the yeast alpha-factor receptor (STE2 gene product) have indicated that oligomerization promotes signaling. Here we have addressed the mechanism by which oligomerization facilitates G protein signaling by examining the ability of ligand binding- and G protein coupling defective alpha-factor receptors to form complexes in vivo and to correct their signaling defects when co-expressed (trans complementation). Newly and previously identified receptor mutants indicated that ligand binding involves the exofacial end of transmembrane domain (TM) 4, whereas G protein coupling involves ic1, ic3, the C-terminal tail, and the intracellular ends of TM2 and TM3. Mutant receptors bearing substitutions in these domains formed homo-oligomeric or hetero oligomeric complexes in vivo, as indicated by results of fluorescence resonance energy transfer experiments. Co-expression of ligand binding- and G protein coupling-defective mutant receptors did not significantly improve signaling. In contrast, co-expression of ic1 and ic3 mutations in trans but not in cis significantly increased signaling efficiency. Therefore, we suggest that subunits of the alpha-factor receptor: 1) are activated independently rather than cooperatively by agonist, and 2) function in a concerted fashion to promote G protein activation, possibly by contacting different subunits or regions of the G protein heterotrimer. PMID- 14764601 TI - On the export of fatty acids from the chloroplast. AB - The model for export of fatty acids from plastids proposes that the acyl-ACP (acyl carrier protein) product of de novo fatty acid synthesis is hydrolyzed in the stroma by acyl-ACP thioesterases and the free fatty acid (FFA) released is then transferred to the outer envelope of the plastid where it is reactivated to acyl-CoA for utilization in cytosolic glycerolipid synthesis. Experiments were performed to assess whether the delivery of nascent FFA from the stroma for long chain acyl-CoA synthesis (LACS) occurs via simple diffusion or a more complex mechanism. The flux through the in vivo FFA pool was estimated using kinetic labeling experiments with spinach and pea leaves. The maximum half-life for FFA in the export pool was < or =1 s. Isolated pea chloroplasts incubated in the light with [14C]acetate gave a linear accumulation of FFA. When CoASH and ATP were present there was also a linear accumulation of acyl-CoA thioesters (plus derived polar lipids), with no measurable lag phase (<30 s), indicating that the FFA pool supplying LACS rapidly reached steady state. The LACS reaction was also measured independently in the dark after in situ generated FFA had accumulated yielding estimates of LACS substrate-velocity relationships. Based on these experiments the LACS reaction with in situ generated FFA as substrate is only about 3% of the LACS activity required in vivo at the very low concentrations of the FFA export pool calculated from the in vivo experiment. Furthermore, bovine serum albumin rapidly removed in situ generated FFA from chloroplasts, but could not compete effectively for "nascent" FFA substrates of LACS. Together the data suggest a locally channeled pool of exported FFA that is closely linked to LACS. PMID- 14764602 TI - Helt, a novel basic-helix-loop-helix transcriptional repressor expressed in the developing central nervous system. AB - Neuronal differentiation is regulated by many basic-helix-loop-helix (bHLH) family transcriptional activators and repressors, and the balance of activity between these factors is important for the differentiation process. Here, we report the identification of a novel transcriptional repressor, designated Helt. Helt encoded a Hey-related bHLH protein containing the bHLH and Orange domains. Helt could homodimerize, and heterodimerize with Hes5 or Hey2. Both the bHLH and Orange domains were involved in the homodimerization. In contrast, only the bHLH domain was required for the heterodimerization with Hey2, whereas only the Orange domain mediated the interaction between Helt and Hes5. Thus, Helt has two dimerization domains, and these domains independently select a partner. Identification of preferred recognition sequences by CASTing experiments revealed that Helt bound to the E box, which was distinct from the Hes1 optimal sequence around the E box core. Not only the core sequence but also sequences flanking the E box were essential for the recognition by Helt and Hes1. Furthermore, Helt repressed transcription from an artificial promoter through binding to the optimal E box elements, as well as transcription from its own promoter. Using in situ hybridization and immunohistochemistry, Helt expression in embryos was investigated. Helt was mainly expressed in undifferentiated neural progenitors in some of the developing brain regions, including the mesencephalon and diencephalon, at the neurogenesis stage. These results suggest that Helt acts as a transcriptional repressor to regulate neuronal differentiation and/or identity. PMID- 14764604 TI - Novel mechanism of PTEN regulation by its phosphatidylinositol 4,5-bisphosphate binding motif is critical for chemotaxis. AB - In chemotaxing cells, localization of phosphatidylinositol 3,4,5-trisphosphate (PI(3,4,5)P3) to the leading edge of the cell sets the direction and regulates the formation of pseudopods at the anterior. We show that the lipid phosphatase activity of PTEN mediates chemotaxis and that the sharp localization of PI(3,4,5)P3 requires localization of PTEN to the rear of the cell. Our data suggest that a phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2) binding motif at the N terminus of PTEN serves the dual role of localizing the enzyme to the membrane and regulating its activity. Mutations in this motif enhance catalytic activity but render the enzyme inactive in vivo by preventing membrane association. The key role of this motif may explain the heretofore puzzling tumor suppressing mutations occurring within the PI(4,5)P2 binding motif. On the other hand, the localization of PTEN does not depend on its phosphatase activity, the actin cytoskeleton, or the intracellular level of PI(3,4,5)P3, suggesting that events controlling localization are upstream of phosphoinositide signaling. PMID- 14764603 TI - Tumor necrosis factor alpha produces insulin resistance in skeletal muscle by activation of inhibitor kappaB kinase in a p38 MAPK-dependent manner. AB - Insulin stimulation produced a reliable 3-fold increase in glucose uptake in primary neonatal rat myotubes, which was accompanied by a similar effect on GLUT4 translocation to plasma membrane. Tumor necrosis factor (TNF)-alpha caused insulin resistance on glucose uptake and GLUT4 translocation by impairing insulin stimulation of insulin receptor (IR) and IR substrate (IRS)-1 and IRS-2 tyrosine phosphorylation, IRS-associated phosphatidylinositol 3-kinase activation, and Akt phosphorylation. Because this cytokine produced sustained activation of stress and proinflammatory kinases, we have explored the hypothesis that insulin resistance by TNF-alpha could be mediated by these pathways. In this study we demonstrate that pretreatment with PD169316 or SB203580, inhibitors of p38 MAPK, restored insulin signaling and normalized insulin-induced glucose uptake in the presence of TNF-alpha. However, in the presence of PD98059 or SP600125, inhibitors of p42/p44 MAPK or JNK, respectively, insulin resistance by TNF-alpha was still produced. Moreover, TNF-alpha produced inhibitor kappaB kinase (IKK) beta activation and inhibitor kappaB-beta and -alpha degradation in a p38 MAPK dependent manner, and treatment with salicylate (an inhibitor of IKK) completely restored insulin signaling. Furthermore, TNF-alpha produced serine phosphorylation of IR and IRS-1 (total and on Ser(307) residue), and these effects were completely precluded by pretreatment with either PD169316 or salicylate. Consequently, TNF-alpha, through activation of p38 MAPK and IKK, produces serine phosphorylation of IR and IRS-1, impairing its tyrosine phosphorylation by insulin and the corresponding activation of phosphatidylinositol 3-kinase and Akt, leading to insulin resistance on glucose uptake and GLUT4 translocation. PMID- 14764605 TI - PDX1, a cellular homeoprotein, binds to and regulates the activity of human cytomegalovirus immediate early promoter. AB - Cellular homeoproteins have been shown to regulate the transcription of several viruses, including herpes simplex viruses, human papillomaviruses, and mouse mammary tumor viruses. Previous studies investigating the anti-viral mechanisms of several cyclin-dependent kinase inhibitors showed that the homeoproteins, pre B-cell leukemia transcription factor 1 (PBX1) and PBX-regulating protein-1 (PREP1), function as transcriptional activators of Moloney murine leukemia virus. Here, we examined the involvement of cellular homeoproteins in regulating the activity of the human cytomegalovirus immediate early (CMV IE) promoter. We identified a 45-bp element located at position -593 to -549 upstream of the transcription start site of the CMV IE gene, which contains multiple putative homeoprotein binding motifs. Gel shift assays demonstrated the physical association between a homeodomain protein, pancreatic-duodenal homeobox factor-1 (PDX1) and the 45-bp cytomegalovirus (CMV) region. We further determined that PDX1 represses the CMV IE promoter activity in 293 cells. Overexpression of PDX1 resulted in a decrease in transcription of the CMV IE gene. Conversely, blocking PDX1 protein synthesis and mutating the PDX1 binding sites enhanced CMV IE dependent transcription. Collectively, our results represent the first work demonstrating that a cellular homeoprotein, PDX1, may be a repressor involved in regulation of human CMV gene expression. PMID- 14764606 TI - Engineering an APRIL-specific B cell maturation antigen. AB - B cell maturation antigen (BCMA) is a tumor necrosis factor receptor family member whose physiological role remains unclear. BCMA has been implicated as a receptor for both a proliferation-inducing ligand (APRIL) and B cell-activating factor (BAFF), tumor necrosis factor ligands that bind to multiple tumor necrosis factor receptor and have been reported to play a role in autoimmune disease and cancer. The results presented herein provide a dual perspective analysis of BCMA binding to both APRIL and BAFF. First, we characterized the binding affinity of monomeric BCMA for its ligands; BAFF binding affinity (IC50 = 8 +/- 5 microm) is about 1000-fold reduced compared with the high affinity interaction of APRIL (IC50 = 11 +/- 3 nm). Second, shotgun alanine scanning of BCMA was used to map critical residues for either APRIL or BAFF binding. In addition to a previously described "DXL" motif (Gordon, N. C., Pan, B., Hymowitz, S. G., Yin, J., Kelley, R. F., Cochran, A. G., Yan, M., Dixit, V. M., Fairbrother, W. J., and Starovasnik, M. A. (2003) Biochemistry 42, 5977-5983), the alanine scanning results predicted four amino acid positions in BCMA (Tyr13, Ile22, Gln25, and Arg27) that could impart ligand specificity. Substitution of Tyr13 was tolerated for BAFF binding but not APRIL binding. Arg27 was required for high affinity binding to APRIL, whereas substitutions of this residue had minimal effect on affinity for BAFF. Further phage display experiments suggested the single mutations of I22K, Q25D, and R27Y as providing the greatest difference in APRIL versus BAFF binding affinity. Incorporation of the Q25D and R27Y substitutions into BCMA produced a dual specificity variant, since it has comparable binding affinity for both APRIL and BAFF, IC50 = 350 and 700 nm, respectively. Binding of the I22K mutant of monomeric BCMA to BAFF was undetectable (IC50 > 100 microm), but affinity for binding to APRIL was similar to wild-type BCMA. Based on these results, a BCMA-Fc fusion with the single I22K mutation was produced that binds APRIL, IC50 = 12 nm, and has no measurable affinity for BAFF. These results suggest that APRIL is the preferred ligand for BCMA and show that specificity can be further modified through amino acid substitutions. PMID- 14764607 TI - SOCS2 induces neurite outgrowth by regulation of epidermal growth factor receptor activation. AB - Suppressor of cytokine signaling (SOCS) 2 is a negative regulator of growth hormone (GH) signaling that regulates body growth postnatally and neuronal differentiation during development. SOCS2 binds to the GH receptor and inhibits GH signaling, including attenuation of STAT5 activation. Here we describe a new function and mechanism of action for SOCS2. Overexpression of SOCS2 in central nervous system neurons promoted neurite outgrowth, and in PC12 cells, neurite outgrowth was induced under nondifferentiating conditions, leading to inhibition of the neurite-inhibitory GTPase Rho and activation of the neurite-promoting GTPase Rac1. Addition of the epidermal growth factor receptor (EGFR) inhibitors PP3 or AG490 or the Src kinase inhibitor PP2 blocked the SOCS2-induced neurite outgrowth. The overexpressed SOCS2 bound to the EGFR, which was constitutively phosphorylated at Tyr845, the Src binding site. Overexpression of the phosphatase SHP-2 reduced the constitutive EGFR phosphorylation and subsequent neurite outgrowth. SOCS2 expression also resulted in a modest 30% decrease in phosphorylation of STAT5b at Tyr699, which is the primary site on STAT5 phosphorylated by GH; however, total tyrosine phosphorylation of STAT5 was decreased by 75-80% under basal and epidermal growth factor-stimulated conditions. Our findings suggest that SOCS2 regulates EGFR phosphorylation, leading to regulation of neurite outgrowth through a novel pathway that is distinct from GH. PMID- 14764608 TI - Interleukin-6 induces expression of Ifi202, an interferon-inducible candidate gene for lupus susceptibility. AB - Systemic lupus erythematosus (SLE) is a prototype autoimmune disease. In human SLE patients, as well as in mouse models of SLE, the development of disease is associated with increased levels of pro-inflammatory cytokines, such as interleukin-6 (IL-6). However, IL-6 target genes contributing to the development of disease remain to be identified. Our previous studies of one mouse model of SLE identified an interferon-inducible gene, Ifi202, as a major contributor to the disease. We now report that IL-6 induces expression of the Ifi202 gene. We found that IL-6 treatment of mouse splenocytes increased levels of Ifi202 mRNA and p202 protein. Furthermore, IL-6 treatment of NIH 3T3 cells or expression of a constitutively active form of STAT3, a known mediator of IL-6 signaling, stimulated the activity of a 202-luc-reporter through a potential STAT3 DNA binding site (the 202-SBS) present in the 5'-regulatory region of the Ifi202 gene. Moreover, treatment of cells with IL-6 stimulated binding of the transcription factor STAT3 to an oligonucleotide containing the 202-SBS in gel mobility shift assays and to the 5'-regulatory region of the Ifi202 gene in chromatin immunoprecipitation assays. Importantly, site-directed mutagenesis of 202-SBS or expression of a dominant negative form of STAT3 significantly reduced constitutive as well as IL-6-stimulated activity of the 202-luc-reporter. Together, our observations support the idea that IL-6 stimulates transcription of the Ifi202 gene through STAT3 activation and predict that increased levels of IL 6 in lupus contribute to up-regulation of p202. PMID- 14764609 TI - Sorting competition with membrane-permeable peptides in intact epithelial cells revealed discrimination of transmembrane proteins not only at the trans-Golgi network but also at pre-Golgi stages. AB - Transmembrane proteins destined to the basolateral cell surface of epithelial cells contain in their cytosolic domain at least two classes of sorting signals: one class promotes exit from the endoplasmic reticulum (ER) and transport to the Golgi complex, and the other class operates at the trans-Golgi network (TGN) specifying segregation into basolateral exocytic pathways. Both kinds of addressing motifs are quite diverse among different proteins. It is unclear to what extent this feature reflects alternative decoding mechanisms or variations in motifs recognized by the same sorting factor. Here we applied a novel strategy based on permeable peptide technology and temperature-sensitive model proteins to study competition between cytosolic sorting motifs in the context of mammalian living cells. We used the transduction domain of HIV-1 Tat protein to make a membrane-permeable peptide of the cytosolic tail of GtsO45, which contains a well characterized ER exit di-acidic (DIE) motif and a tyrosine-based basolateral sorting signal (YTDI). This peptide added to the media inhibited transport of GtsO45 from both ER-to-Golgi and TGN-to-basolateral cell surface in transfected Madin-Darby canine kidney cells. Instead, it did not affect the exocytic trafficking of a GtsO45-derived chimeric protein bearing 30 juxtamembrane residues from the cytosolic domain of the epidermal growth factor receptor that contains a variant ER exit motif (ERE) and an unconventional proline-based basolateral sorting signal. These results not only proved the feasibility of competing for sorting events in intact cells but also showed that distinct plasma membrane proteins can be discriminated at pre-TGN stages, and that basolateral sorting involves different recognition elements for tyrosine-based motifs and an unconventional basolateral motif. PMID- 14764610 TI - Development of a provisional domain model for the nursing process for use within the Health Level 7 reference information model. AB - OBJECTIVE: Since 1999, the Nursing Terminology Summits have promoted the development, evaluation, and use of reference terminology for nursing and its integration into comprehensive health care data standards. The use of such standards to represent nursing knowledge, terminology, processes, and information in electronic health records will enhance continuity of care, decision support, and the exchange of comparable patient information. As part of this activity, working groups at the 2001, 2002, and 2003 Summit Conferences examined how to represent nursing information in the Health Level 7 (HL7) Reference Information Model (RIM). DESIGN: The working groups represented the nursing process as a dynamic sequence of phases, each containing information specific to the activities of the phase. They used Universal Modeling Language (UML) to represent this domain knowledge in models. An Activity Diagram was used to create a dynamic model of the nursing process. After creating a structural model of the information used at each stage of the nursing process, the working groups mapped that information to the HL7 RIM. They used a hierarchical structure for the organization of nursing knowledge as the basis for a hierarchical model for "Findings about the patient." The modeling and mapping reported here were exploratory and preliminary, not exhaustive or definitive. The intent was to evaluate the feasibility of representing some types of nursing information consistently with HL7 standards. MEASUREMENTS: The working groups conducted a small-scale validation by testing examples of nursing terminology against the HL7 RIM class "Observation." RESULTS: It was feasible to map patient information from the proposed models to the RIM class "Observation." Examples illustrate the models and the mapping of nursing terminology to the HL7 RIM. CONCLUSION: It is possible to model and map nursing information into the comprehensive health care information model, the HL7 RIM. These models must evolve and undergo further validation by clinicians. The integration of nursing information, terminology, and processes in information models is a first step toward rendering nursing information machine-readable in electronic patient records and messages. An eventual practical result, after much more development, would be to create computable, structured information for nursing documentation. PMID- 14764611 TI - An enriched unified medical language system semantic network with a multiple subsumption hierarchy. AB - OBJECTIVE: The Unified Medical Language System's (UMLS's) Semantic Network's (SN's) two-tree structure is restrictive because it does not allow a semantic type to be a specialization of several other semantic types. In this article, the SN is expanded into a multiple subsumption structure with a directed acyclic graph (DAG) IS-A hierarchy, allowing a semantic type to have multiple parents. New viable IS-A links are added as warranted. DESIGN: Two methodologies are presented to identify and add new viable IS-A links. The first methodology is based on imposing the characteristic of connectivity on a previously presented partition of the SN. Four transformations are provided to find viable IS-A links in the process of converting the partition's disconnected groups into connected ones. The second methodology identifies new IS-A links through a string matching process involving names and definitions of various semantic types in the SN. A domain expert is needed to review all the results to determine the validity of the new IS-A links. RESULTS: Nineteen new IS-A links are added to the SN, and four new semantic types are also created to support the multiple subsumption framework. The resulting network, called the Enriched Semantic Network (ESN), exhibits a DAG-structured hierarchy. A partition of the ESN containing 19 connected groups is also derived. CONCLUSION: The ESN is an expanded abstraction of the UMLS compared with the original SN. Its multiple subsumption hierarchy can accommodate semantic types with multiple parents. Its representation thus provides direct access to a broader range of subsumption knowledge. PMID- 14764612 TI - Understanding implementation: the case of a computerized physician order entry system in a large Dutch university medical center. AB - Most studies of the impact of information systems in organizations tend to see the implementation process as a "rollout" of technology, as a technical matter removed from organizational dynamics. There is substantial agreement that the success of implementing information systems is determined by organizational factors. However, it is less clear what these factors are. The authors propose to characterize the introduction of an information system as a process of mutual shaping. As a result, both the technology and the practice supported by the technology are transformed, and specific technical and social outcomes gradually emerge. The authors suggest that insights from social studies of science and technology can help to understand an implementation process. Focusing on three theoretical aspects, the authors argue first that the implementation process should be understood as a thoroughly social process in which both technology and practice are transformed. Second, following Orlikowski's concept of "emergent change," they suggest that implementing a system is, by its very nature, unpredictable. Third, they argue that success and failure are not dichotomous and static categories, but socially negotiated judgments. Using these insights, the authors have analyzed the implementation of a computerized physician order entry (CPOE) system in a large Dutch university medical center. During the course of this study, the full implementation of CPOE was halted, but the aborted implementation exposed issues on which the authors did not initially focus. PMID- 14764613 TI - A simple and practical dictionary-based approach for identification of proteins in Medline abstracts. AB - OBJECTIVE: The aim of this study was to develop a practical and efficient protein identification system for biomedical corpora. DESIGN: The developed system, called ProtScan, utilizes a carefully constructed dictionary of mammalian proteins in conjunction with a specialized tokenization algorithm to identify and tag protein name occurrences in biomedical texts and also takes advantage of Medline "Name-of-Substance" (NOS) annotation. The dictionaries for ProtScan were constructed in a semi-automatic way from various public-domain sequence databases followed by an intensive expert curation step. MEASUREMENTS: The recall and precision of the system have been determined using 1000 randomly selected and hand-tagged Medline abstracts. RESULTS: The developed system is capable of identifying protein occurrences in Medline abstracts with a 98% precision and 88% recall. It was also found to be capable of processing approximately 300 abstracts per second. Without utilization of NOS annotation, precision and recall were found to be 98.5% and 84%, respectively. CONCLUSION: The developed system appears to be well suited for protein-based Medline indexing and can help to improve biomedical information retrieval. Further approaches to ProtScan's recall improvement also are discussed. PMID- 14764615 TI - Audience response made easy: using personal digital assistants as a classroom polling tool. AB - Both teachers and students benefit from an interactive classroom. The teacher receives valuable input about effectiveness, student interest, and comprehension, whereas student participation, active learning, and enjoyment of the class are enhanced. Cost and deployment have limited the use of existing audience response systems, allowing anonymous linking of teachers and students in the classroom. These limitations can be circumvented, however, by use of personal digital assistants (PDAs), which are cheaper and widely used by students. In this study, the authors equipped a summer histology class of 12 students with PDAs and wireless Bluetooth cards to allow access to a central server. Teachers displayed questions in multiple-choice format as a Web page on the server and students responded with their PDAs, a process referred to as polling. Responses were immediately compiled, analyzed, and displayed. End-of-class survey results indicated that students were enthusiastic about the polling tool. The surveys also provided technical feedback that will be valuable in streamlining future trials. PMID- 14764616 TI - A frequency-based technique to improve the spelling suggestion rank in medical queries. AB - OBJECTIVE: There is an abundance of health-related information online, and millions of consumers search for such information. Spell checking is of crucial importance in returning pertinent results, so the authors propose a technique for increasing the effectiveness of spell-checking tools used for health-related information retrieval. DESIGN: A sample of incorrectly spelled medical terms was submitted to two different spell-checking tools, and the resulting suggestions, derived under two different dictionary configurations, were re-sorted according to how frequently each term appeared in log data from a medical search engine. MEASUREMENTS: Univariable analysis was carried out to assess the effect of each factor (spell-checking tool, dictionary type, re-sort, or no re-sort) on the probability of success. The factors that were statistically significant in the univariable analysis were then used in multivariable analysis to evaluate the independent effect of each of the factors. RESULTS: The re-sorted suggestions proved to be significantly more accurate than the original list returned by the spell-checking tool. The odds of finding the correct suggestion in the number one rank were increased by 63% after re-sorting using the authors' method. This effect was independent of both the dictionary and the spell-checking tools that were used. CONCLUSION: Using knowledge about the frequency of a given word's occurrence in the medical domain can significantly improve spelling correction for medical queries. PMID- 14764617 TI - Training the next generation of informaticians: the impact of "BISTI" and bioinformatics--a report from the American College of Medical Informatics. AB - In 2002-2003, the American College of Medical Informatics (ACMI) undertook a study of the future of informatics training. This project capitalized on the rapidly expanding interest in the role of computation in basic biological research, well characterized in the National Institutes of Health (NIH) Biomedical Information Science and Technology Initiative (BISTI) report. The defining activity of the project was the three-day 2002 Annual Symposium of the College. A committee, comprised of the authors of this report, subsequently carried out activities, including interviews with a broader informatics and biological sciences constituency, collation and categorization of observations, and generation of recommendations. The committee viewed biomedical informatics as an interdisciplinary field, combining basic informational and computational sciences with application domains, including health care, biological research, and education. Consequently, effective training in informatics, viewed from a national perspective, should encompass four key elements: (1). curricula that integrate experiences in the computational sciences and application domains rather than just concatenating them; (2). diversity among trainees, with individualized, interdisciplinary cross-training allowing each trainee to develop key competencies that he or she does not initially possess; (3). direct immersion in research and development activities; and (4). exposure across the wide range of basic informational and computational sciences. Informatics training programs that implement these features, irrespective of their funding sources, will meet and exceed the challenges raised by the BISTI report, and optimally prepare their trainees for careers in a field that continues to evolve. PMID- 14764618 TI - Genetics of familial combined hyperlipidemia and risk of coronary heart disease. AB - Coronary heart disease is the leading cause of death in developed countries. This alarming statistic is partly attributable to lifestyle, and partly due to the genetic factors that make humans highly susceptible to atherosclerotic vascular disease. The principal metabolic causes of atherosclerosis include hyperlipidemia, hypertension, obesity, insulin resistance and diabetes mellitus. Here we discuss the aetiology of familial combined hyperlipidemia (FCHL), a highly atherogenic disorder affecting 1-2% of the Western world. Genome-wide linkage studies indicate that more than three genes contribute to the pernicious lipid profile of FCHL, and that these genes reside within the 1q21-23, 11p14.1 q12.1 and 16q22-24.1 chromosomal regions. Other loci include 1p31, 6q16.1-16.3 and 8p23.3-22, but the linkage data for these are not yet persuasive. Combined linkage and association analyses provide compelling evidence for the involvement of two distinct alleles at the APOA1/C3/A4/A5 gene cluster in the transmission of FCHL. An important lesson arising from the study of a complex genetic disorder, such as FCHL, that lacks a consensus on diagnostic criteria, is that an understanding of complex genetic disorders can derive from comparative analyses of genome-wide linkage data generated from conditions that share phenotypic overlap. The identification of potential genetic overlap between FCHL and the Metabolic Syndrome, which is estimated to affect 47 million Americans, promises to deliver new targets for reducing the risk of important conditions such as cardiovascular disease and stroke. PMID- 14764619 TI - Implications of human genome architecture for rearrangement-based disorders: the genomic basis of disease. AB - The term 'genomic disorder' refers to a disease that is caused by an alteration of the genome that results in complete loss, gain or disruption of the structural integrity of a dosage sensitive gene(s). In most of the common chromosome deletion/duplication syndromes, the rearranged genomic segments are flanked by large (usually >10 kb), highly homologous low copy repeat (LCR) structures that can act as recombination substrates. Recombination between non-allelic LCR copies, also known as non-allelic homologous recombination, can result in deletion or duplication of the intervening segment. Recent findings suggest that other chromosomal rearrangements, including reciprocal, Robertsonian and jumping translocations, inversions, isochromosomes and small marker chromosomes, may also involve susceptibility to rearrangement related to genome structure or architecture. In several cases, LCRs, AT-rich palindromes and pericentromeric repeats are located at such rearrangement breakpoints. Analysis of the products of recombination at the junctions of the rearrangements reveals both homologous recombination and non-homologous end joining as causative mechanisms. Thus, a more global concept of genomic disorders emerges in which susceptibility to rearrangements occurs due to underlying complex genomic architecture. Interestingly, this architecture plays a role not only in disease etiology, but also in primate genome evolution. In this review, we discuss recent advances regarding general mechanisms for the various rearrangements of our genome, and potential models for rearrangements with non-homologous breakpoint regions. PMID- 14764620 TI - Genetic dissection of myocilin glaucoma. AB - Primary open-angle glaucoma (POAG) is a complex disease with unknown causes. However, in the past decade, POAG has been linked to six chromosomal regions, of which the gene MYOC encoding myocilin and the gene OPTN encoding optineurin have been identified to harbor causal mutations (disease-causing variants, DCV). POAG caused by DCV at MYOC has been termed "myocilin glaucoma". Clinically, DCV at MYOC may manifest as a typical POAG, normal tension glaucoma, or ocular hypertension without glaucoma. Individuals with the Arg46Stop mutation that almost knocks out the entire coding sequence may have severe glaucoma or no glaucoma. Genetically, myocilin glaucoma follows autosomal dominant, recessive or no pattern of inheritance. DCV at MYOC cause POAG in interaction with environmental factors and DCV at other loci. Most DCV at MYOC are relatively young, and the Gln368Stop mutation is exclusively European in origin. The overall frequency of DCV at MYOC is similar among African, Caucasian and Asian probands with POAG. Because of this fact and the higher prevalence of POAG in African descendants compared with Caucasians or Asians, the overall frequency of DCV at MYOC is several-fold higher in the general population of African descendants, which is in part responsible for their higher prevalence of POAG. Although the Arg46Stop mutation was often observed in normal controls, Arg46Stop carriers tend to have higher risk of developing POAG. Polymorphisms at several loci including MYOC are associated with POAG, and play an important role in the pathogenesis of POAG. PMID- 14764621 TI - Patterns of ethnic diversity among the genes that influence AIDS. AB - Although HIV-1/AIDS emerged late in the last century, more than 42 million individuals have been infected and 25 million have died worldwide, making AIDS, like malaria, a strong selective force for disease-associated genetic factors. Many of the genes that mediate immune response or that are co-opted by HIV-1 for completion of its lifecycle show differences in allele frequencies, as a result of drift, migration or selection. Here we show that the majority of AIDS candidate genes and AIDS restriction genes show significant differences in allele frequencies, possibly the result of historic selective pressures. These genes are undergoing present day natural selection in populations with high AIDS prevalence. PMID- 14764622 TI - The genetic basis of systemic lupus erythematosus--knowledge of today and thoughts for tomorrow. AB - Systemic lupus erythematosus (SLE) is a chronic rheumatic disease with an autoimmune etiology. Nuclear components of the cells are the main targets of the autoimmune reaction, affecting virtually any organ in the body. SLE is also called a prototype disease due to a substantial overlap in its clinical symptoms with other autoimmune diseases. Therefore the understanding of the mechanisms underlying SLE may contribute to advances in studies and development of new treatments for several autoimmune diseases. SLE is a complex disease with both genetic factors (mutations or susceptibility alleles) and environmental factors (infections, drugs, stress, exposures, etc.) contributing to its development. In this article we will give an overview of the latest findings in genetics of SLE, concentrating on the two most interesting and promising pathways: the PD-1 and the interferon pathways. PMID- 14764623 TI - Alzheimer's disease: one disorder, too many genes? AB - The research of Alzheimer's disease (AD) genetics has been extremely prolific over the past decade, and currently more than 10 genes are reported to show either positive or negative evidence for disease association per month. Here, we review all 90 studies from 2003 reporting a total of 127 association findings between candidate genes and AD. While most positive results were largely contradictory, we identified three loci-on chromosomes 6p21, 10q24, 11q23-that yielded positive results in three or more independent studies, in addition to the well-established AD association with the gene encoding apolipoprotein E (APOE). Based on these data, we suggest that it may be prudent for investigators to pay closer attention to issues such as power, replicability and haplotype structure prior to initial publication. This should serve to greatly decrease the likelihood of false positive and false negative findings reported in future years. PMID- 14764624 TI - Genetics of essential hypertension. AB - Essential hypertension affects 1 billion people worldwide and its genetic basis is well established. For this review we surveyed the literature on the genetics of hypertension during the past 18 months and we now report the highlights. There has been publication of the two largest genome scans for blood pressure and new loci including significant linkage to chromosome 6q have been reported. The molecular basis of Gordon's syndrome has been partially unravelled with a dual function for WNK4 in ion transport regulation being discovered. There has also been progress in narrowing rodent quantitative trait loci using congenic approaches and several linkage peaks have now been demonstrated to have more than one loci. We also report some of the initial findings from pharmacogenetic studies. PMID- 14764625 TI - Genetics of inflammatory bowel disease: progress and prospects. AB - Strong epidemiological evidence for a genetic contribution to pathogenesis in inflammatory bowel disease (IBD) has stimulated efforts to identify susceptibility genes for both of its major clinical forms, Crohn's disease and ulcerative colitis. Genome scans for linkage have indicated multiple regions of interest, but replication of these has been limited. The detection of linkage on chromosome 16 (IBD1) led to the unequivocal identification of the NOD2 gene (now called CARD15) as a susceptibility gene for Crohn's disease. This seminal discovery has provided proof of principle for positional cloning and candidate gene approaches to identify IBD genes. It has also led to useful strategic insights in complex disease genetics, and generated new directions in the investigation of the molecular pathways to pathogenesis. Linkage and association studies have also provided strong support for IBD susceptibility genes on chromosomes 5q31, 6p21 and 19p, while loci of interest at 3p, 3q and 14q require further follow-up. Although important obstacles to further progress will need to be overcome, the successes of the past 2 years suggest that a detailed description of the genetic basis of inflammatory bowel disease is a realistic goal. PMID- 14764626 TI - Asthma genetics 2003. AB - The use of positional cloning for the identification of complex trait susceptibility genes has gained momentum with the completion of the human genome project. The approach involves the collection of well-phenotyped cohorts (either family-based or case-control designs), the generation of high-density single nucleotide polymorphism linkage disequilibrium maps, and the application of powerful statistical methods to localize narrow regions of genetic association with disease. In 2003, two novel genes relating to asthma were identified using this approach, PHF11 and DPP10, neither of which had previously been implicated in the pathobiology of either asthma or allergy. In addition, further support for ADAM33 (the first asthma susceptibility gene identified by positional cloning) as an asthma gene was presented, although with mixed results. These discoveries open new avenues for research in asthma and allergy, and highlight the power (and limitations) of positional cloning for the identification of asthma genes, and complex trait genes in general. PMID- 14764627 TI - Cocaine- and amphetamine-regulated transcript peptide in the rat anterior pituitary gland is localized in gonadotrophs and suppresses prolactin secretion. AB - Cocaine- and amphetamine-regulated transcript (CART) mRNA and CART peptide are abundant in the hypothalamic nuclei that control anterior pituitary function. CART peptide has also been localized in the anterior pituitary gland itself, although its role in pituitary function has not as yet been elucidated. In the present study, we investigated the localization and function of CART peptide in the anterior pituitary gland. Immunohistochemical observations revealed that CART peptide colocalized with FSH and LH in gonadotroph cells but that it was absent from the other hormone-producing cells. Immunoelectronmicroscopy suggested that CART peptide and gonadotropin were colocalized in the same secretory granules. CART peptide suppressed prolactin release from dispersed anterior pituitary cells 15 min after its addition into the media [basal production, 234.9 +/- 14.6 ng/ml vs. CART 55-102 peptide 100 nm, 125.0 +/- 18.2 ng/ml (P < 0.01, n = 5)]. Prolactin release was suppressed by CART in a dose-related manner; on the other hand, CART peptide did not affect the secretion of other anterior pituitary hormones. CART peptide synthesis by these cells was elevated 15 min after the addition of leptin to the media (100 nm), as determined by immunoblotting, but LHRH (10 nm) did not significantly affect CART peptide expression. These findings suggest that CART synthesis in the anterior pituitary may be stimulated by leptin and that CART peptide may play a role in the regulation of anterior pituitary hormone secretion in the rat. PMID- 14764628 TI - The 26S proteasome participates in the sequential inhibition of estrous behavior induced by progesterone in rats. AB - Estrous behavior induced by progesterone (P) treatment of estradiol-primed rats is followed by a period in which females do not respond behaviorally to a second administration of P [sequential inhibition (SI)]. SI is thought to involve P dependent down-regulation of hypothalamic P receptor (PR) content. This study tested the hypothesis that the 26S proteasome participates in the regulation of SI and brain PR content in female rats. Ovariectomized, estrogen-primed (estradiol benzoate, 2 microg s.c.) adult rats were injected with P (1 mg s.c.) alone or P with the proteasome inhibitors Z-Ile-Glu (OBu(1))-Ala-Leu-H (PSI, 300 microg/100 g s.c.) or N alpha-tosyl-lysyl chloromethyl ketone (TLCK, 200 microg i.p.) administered 48 h after estradiol priming. Sexual behavior was assessed in all animals 4 h later. These two agents inhibit 26S proteasome-mediated protein degradation by different mechanisms. To explore SI, the animals received a second P injection 24 h after the first, and a second sexual behavior test was performed 4 h later. After this test, brains were excised, and proteins were extracted from the preoptic area and the hypothalamus and processed for semiquantitative immunoblotting. In the first sexual behavior test (facilitation test), all animals treated with estradiol + P exhibited intense lordosis behavior. In the second sexual behavior test (inhibition test), both lordosis and proceptivity were significantly reduced in response to the second administration of P (SI). The magnitude of SI was significantly attenuated by the administration of either PSI or TLCK concurrently with the first P injection. The first P injection reduced PR content in the hypothalamus but not in the preoptic area. In contrast, PSI and TLCK significantly increased PR content in both structures. Our results suggest that PR degradation by the 26S proteasome participates in the expression of P-induced SI in female rats. PMID- 14764629 TI - Role of signal transducer and activator of transcription 3 in regulation of hypothalamic trh gene expression by leptin. AB - During starvation in rodents, the hypothalamic-pituitary-thyroid axis is down regulated, resulting in low circulating thyroid hormone levels. This involves a reduction in hypothalamic TRH mRNA that is caused in part by a fall in serum leptin levels, which is sensed by neurons within the hypothalamus. The mechanism by which this regulation occurs is not fully understood. Here we show transfection data and in vivo evidence, suggesting that leptin can regulate trh gene expression via activation of intracellular signal transducer and activator of transcription 3 (STAT3) proteins in TRH neurons. In trh promoter assays using transfected cells, functional STAT3 proteins are required for maximal activation of the trh promoter by leptin. Consistent with this, the STAT3-binding site on the leptin receptor is also required for this regulation. Using double immunohistochemistry, we show that peripherally administered leptin rapidly stimulates STAT3 phosphorylation in approximately 40% of TRH neurons in the paraventricular nucleus of the hypothalamus (PVN) in rats. Detailed anatomical analyses reveal that the leptin-responsive TRH neurons are concentrated in the caudal region of the medial and periventricular parvocellular subnucleus of the PVN. Combined, our data show that only a subpopulation of TRH neurons in the PVN is leptin responsive and suggest that stimulation of hypothalamic trh gene expression by leptin involves activation of STAT3 and that this signaling pathway is important for regulation of the hypothalamic-pituitary-thyroid axis by leptin. PMID- 14764630 TI - Leptin signaling targets the thyrotropin-releasing hormone gene promoter in vivo. AB - The regulation of TRH gene expression in the paraventricular nucleus of the hypothalamus (PVH) by leptin is critical for normal function of the thyroid axis in rodents and humans. The TRH neuron in the PVH expresses both leptin and melanocortin-4 receptors, suggesting that both signaling systems may regulate TRH gene expression in vivo. Indeed, the TRH promoter responds to both of these signaling pathways in cell culture through identified cis-acting elements, which include signal transducer and activator of transcription (STAT) 3 and cAMP response element binding protein binding sites that mediate leptin and melanocortin responses, respectively. To determine whether leptin signaling can directly target the TRH promoter in vivo, we developed a chromatin immunoprecipitation assay to use on leptin-treated animals. After a single injection of leptin in fasting animals, we could detect a significant increase in TRH gene expression in the PVH that correlated well with the induction of phosphorylated-STAT3 in the hypothalamus. Furthermore, using a STAT3 antibody, we could immunoprecipitate the STAT-binding site containing regions of both the TRH promoter and the promoter of the suppressor of cytokine signaling-3 gene, another well-defined target of leptin action. In contrast, upstream regions of these promoters that lack STAT sites were not precipitated. Taken together these experiments demonstrate that STAT3 mediates transcriptional effects of leptin in vivo and that the TRH promoter is a likely direct site of leptin action. In addition, these experiments demonstrate that chromatin immunoprecipitation can be used to characterize leptin-signaling in vivo. PMID- 14764631 TI - Cloning and characterization of granulosa cell high-mobility group (HMG)-box protein-1, a novel HMG-box transcriptional regulator strongly expressed in rat ovarian granulosa cells. AB - Specific events in the ovary are dependent on gene expression in the tissue. By screening a rat ovarian granulosa cell cDNA library, a cDNA clone encoding a novel transcription factor-like protein containing a high-mobility group-box, referred to as granulosa cell high-mobility group-box protein-1 (GCX-1), was identified. The expression of GCX-1 is restricted to the hypothalamus, pituitary, testis, uterus, and ovary but was not detected in the adrenal gland. An in situ hybridization study revealed that the expression of GCX-1 was restricted to granulosa cell layers in early-stage follicles, and the expression was very low in large antral follicles and the corpus luteum, but localized expression in the testis or pituitary was not clear. Endogenous GCX-1 protein in the granulosa cells was identified by a Western blot analysis, and an analysis using the green fluorescence protein-GCX-1 fusion protein revealed that the GCX-1 protein was localized in the cell nucleus. GAL4 fusion protein-based assays demonstrated that GCX-1 is a potent transcriptional activator, and its putative transactivation domain was mapped to the region between amino acid residues 25 and 63 from the N terminus. These data strongly suggest that GCX-1 is likely a novel transcriptional activator that is exclusively expressed in reproductive tissues involving the hypothalamo-pituitary-gonadal axis, and functions as a specific regulator of follicular development, and may also participate in other specific events related to reproduction, particularly in the female. PMID- 14764632 TI - Orexin 1 receptor messenger ribonucleic acid expression and stimulation of testosterone secretion by orexin-A in rat testis. AB - Orexins are hypothalamic neuropeptides primarily involved in the regulation of food intake and arousal states. In addition, a role for orexins as central neuroendocrine modulators of reproductive function has recently emerged. Prepro orexin and orexin type-1 receptor mRNAs have been detected in the rat testis. This raises the possibility of additional peripheral actions of orexins in the control of reproductive axis, which remains so far unexplored. To analyze the biological effects and mechanisms of action of orexins in the male gonad, we evaluated testicular expression of orexin receptor 1 (OX(1)R) and orexin receptor 2 (OX(2)R) mRNAs in different experimental settings and the effect of orexin-A on testicular testosterone (T) secretion. Persistent expression of OX(1)R mRNA was demonstrated in the rat testis throughout postnatal development. In contrast, OX(2)R transcript was not detected at any developmental stage. Expression of OX(1)R mRNA persisted after selective elimination of mature Leydig cells and was detected in isolated seminiferous tubules at defined stages of the seminiferous epithelial cycle. In addition, testicular OX(1)R mRNA expression appeared to be under hormonal regulation; it was reduced by long-term hypophysectomy and partially restored by FSH replacement, whereas down-regulation was observed after exposure to increasing doses of the ligand in vitro. Moreover, OX(1)R mRNA expression was sensitive to neonatal imprinting by estrogen. Finally, orexin-A, in a dose-dependent manner, significantly increased basal, but not human choriogonadotropin-stimulated, T secretion in vitro. A similar stimulatory effect was observed in vivo after intratesticular administration of orexin-A. In conclusion, our present results provide the first evidence for the regulated expression of OX(1)R mRNA and functional role of orexin-A in the rat testis. Overall, our data are suggestive of a novel site of action of orexins in the control of male reproductive axis. PMID- 14764633 TI - Dynamics of thyroid-stimulating and -blocking antibodies to the thyrotropin receptor in a murine model of Graves' disease. AB - Graves' disease is characterized by the presence of autoantibodies to the TSH receptor (TSHR). There are multiple antibodies to the TSHR, with thyroid stimulating antibodies (TSAbs) and TSH-stimulating blocking antibodies (TSBAbs), which in patients can fluctuate over time, resulting in changes in disease activity. Recently, animal models of Graves' disease have been developed, but it is not known whether the induced TSAbs and TSBAbs change spontaneously with time to influence disease. We used fibroblasts expressing major histocompatability complex (MHC) class II and human TSHR murine model to study anti-TSHR antibody patterns in serial bleeds of 23 animals. Anti-TSHR antibody responses were first detectable after 7-8 wk of first immunization. Moreover, the pattern of the TSAbs or TSBAbs was selected early in the response. The majority of the animals showed anti-TSHR antibodies that were either TSAb or TSBAb responses and were maintained throughout the course of 17-24 wk of the experiment. Remarkably, a proportion of mice (13%) displayed presence of antibodies with both TSAbs and TSBAbs, which appeared to cycle over time and thus mimic the fluctuations described in some hyperthyroid patients. Analyses of the linear epitopes to TSHR by peptide scanning showed that there was no early restricted epitope response. Thus, despite using an inbred strain, the initial response appears to target different regions of the receptor in different animals. Our data show that anti-TSHR antibody epitopes in the model display heterogeneity in TSHR epitopes, which vary in individual animals as well as in their regulation. PMID- 14764634 TI - nitric oxide triggers the phosphatidylinositol 3-kinase/Akt survival pathway in insulin-producing RINm5F cells by arousing Src to activate insulin receptor substrate-1. AB - Mechanisms involved in the protective action of nitric oxide (NO) in insulin producing cells are a matter of debate. We have previously shown that pharmacological inhibition of c-Src cancels the antiapoptotic action of low and sustained concentrations of exogenous NO. In this study, using insulin-producing RINm5F cells that overexpress Src either permanently active (v-Src) or dominant negative (dn-Src) forms, we determine that this tyrosine kinase is the principal mediator of the protective action of NO. We also show that Src-directed activation of insulin receptor substrate-1, phosphatidylinositol 3-kinase (PI3K), Akt, and Bad phosphorylation conform a substantial component of the survival route because pharmacological inhibition of PI3K and Akt canceled the antiapoptotic effects of NO. Studies performed with the protein kinase G (PKG) inhibitor KT-5823 revealed that NO-dependent activation of c-Src/ insulin receptor substrate-1 is not affected by PKG activation. By contrast, Akt and Bad activation are partially dependent on PKG activation. Endogenous production of NO after overexpression of endothelial nitric oxide synthase in RINm5F cells mimics the effects produced by generation of low amounts of NO from exogenous diethylenetriamine/NO. In addition, we found that NO produces c-Src/PI3K- and PKG dependent activation of ERK 1/2. The MAPK kinase inhibitor PD 98059 suppresses NO dependent protection from DNA fragmentation induced by serum deprivation. The protective action of low and sustained concentration of NO is also observed in staurosporine- and Taxol-induced apoptosis. Finally, NO also protects isolated rat islets from DNA fragmentation induced by serum deprivation. These data strengthen the notion that NO production at physiological levels plays a role in protection from apoptosis in pancreatic beta-cells. PMID- 14764635 TI - Interaction of tumor necrosis factor-alpha- and thiazolidinedione-regulated pathways in obesity. AB - Thiazolidinediones (TZDs) are potent insulin-sensitizing compounds and high affinity ligands for the transcription factor peroxisomal proliferator-activated receptor gamma. The mechanism through which TZDs improve insulin sensitivity, however, is not clear. In this study, we asked whether the ability of TZD to suppress and antagonize TNF alpha is an underlying mechanism for its molecular and physiological effects, using obese (ob/ob) mice lacking TNF alpha function. We found that the lipid-lowering effects of TZD are completely independent of TNF alpha suppression, and the insulin-sensitizing effects of TZD are partially independent. TZD treatment improved insulin sensitivity in ob/ob mice both with and without functional TNF alpha, albeit with different absolute potency. To characterize the potential interdependency of TZD- and TNF alpha-regulated pathways at the molecular level, we also performed four-way transcriptional profiling of white adipose tissue of TZD- and vehicle-treated ob/ob mice, with and without TNF alpha function. The majority of metabolic genes identified were regulated independent of the presence of TNF alpha, whereas most effects on inflammatory mediators were dependent on TNF alpha. This study demonstrates that the insulin-sensitizing action of TZD occurs partially through TNF-independent mechanisms, although a subset of the molecular effects of TZD treatment in adipose tissue depends on TNF alpha. PMID- 14764636 TI - Transgenic mice producing major histocompatibility complex class II molecules on thyroid cells do not develop apparent autoimmune thyroid diseases. AB - The expression of major histocompatibility complex (MHC) class II molecules on thyrocytes has been demonstrated in autoimmune thyroid diseases. However, the role of this aberrant MHC class II in disease development is controversial. In particular, it remains unknown whether MHC class II expression on thyrocytes, which are nonprofessional antigenpresenting cells, plays a role in inducing autoimmune processes. To clarify this issue, we have produced transgenic mice harboring an MHC class II gene ligated to the promoter of the rat TSH receptor. We obtained three lines of transgenic mice, and the expression of MHC class II by the thyrocytes was demonstrated by immunofluorescence staining and flow cytometry. Our examination revealed no obvious abnormalities in thyroid histology or in thyroid autoantibody production in these transgenic mice. Although serum free T(4) levels were slightly lower than those of their nontransgenic littermates, no transgenic mouse suffered from clinical hypothyroidism or hyperthyroidism. Furthermore, thyroid lymphocytic infiltration was absent, and MHC class II-expressing thyrocytes obtained from transgenic mice failed to stimulate the proliferation of autologous T cells in vitro. Taken together, these results show that transgenic mice with MHC class II molecules on their thyrocytes do not develop apparent autoimmune thyroid diseases, suggesting that aberrant MHC class II expression alone is not sufficient to induce thyroid autoimmunity. PMID- 14764637 TI - Androgens regulate phosphodiesterase type 5 expression and functional activity in corpora cavernosa. AB - By real-time RT-PCR and Western blot analysis, we found that phosphodiesterase type 5 (PDE5) mRNA and protein abundance was several fold higher in human male than in female reproductive tracts. The highest mRNA level (>1 x 10(7) molecules/microg total RNA) was detected in human corpora cavernosa (CC), where PDE5 protein was immunolocalized in both muscular and endothelial compartment. The possible role of androgens in regulating PDE5 expression was studied using a previously established rabbit model of hypogonadotropic hypogonadism. In this model, hypogonadism reduced, and testosterone (T) supplementation restored, CC PDE5 gene and protein expression. In addition, T supplementation completely rescued and even enhanced cyclic GMP conversion to metabolites, without changing IC(50) for sildenafil (IC(50) = 2.16 +/- 0.62 nm). In control CC strips, sildenafil dose-dependently increased relaxation induced by electrical field stimulation, with EC(50) = 3.42 +/- 1.7 nm. Hypogonadism reduced, and T increased, sildenafil effect on electrical field stimulation, again without changing their relative EC(50) values. CC sensitivity to the NO-donor NCX4040 was greater in hypogonadal rabbit strips than in control or T-treated counterparts. Moreover, sildenafil enhanced NCX4040 effect in eugonadal rabbit strips but not in hypogonadal ones. This suggests that androgens up-regulate PDE5 in rabbit penis. We also measured PDE5 gene expression and metabolic activity in human CC from male-to-female transsexual individuals, chronically treated with estrogens and cyproterone acetate. Comparing the observed values vs. eugonadal controls, PDE5 mRNA, protein, and functional activity were significantly reduced. In conclusion, we demonstrated, for the first time, that androgens positively regulate PDE5, thus providing a possible explanation about the highest abundance of this enzyme in male genital tract. PMID- 14764638 TI - Acetylcholine is an autocrine or paracrine hormone synthesized and secreted by airway bronchial epithelial cells. AB - The role of acetylcholine (ACh) as a key neurotransmitter in the central and peripheral nervous system is well established. However, the role of ACh may be broader because ACh may also function as an autocrine or paracrine signaling molecule in a variety of nonneuronal tissues. To begin to establish ACh of nonneuronal origin as a paracrine hormone in lung, we have examined neonatal and adult monkey bronchial epithelium for the components involved in nicotinic cholinergic signaling. Using immunohistochemistry and RT-PCR, we have demonstrated in lung bronchial epithelial cells (BECs) expression of choline acetyltransferase, the vesicular ACh transporter, the choline high-affinity transporter, alpha7, alpha4, and beta2 nicotinic ACh receptor (nAChR) subunits, and the nAChR accessory protein lynx1. Confocal microscopy demonstrates that these factors are expressed in epithelial cells and are clearly distinct from neighboring nerve fibers. Confirmation of RNA identity has been confirmed by partial sequence analysis of PCR products and by cDNA cloning. Primary culture of BECs confirms the synthesis and secretion of ACh and the activity of cholinesterases. Thus, ACh meets all the criteria for an autocrine/paracrine hormone in lung bronchial epithelium. The nonneuronal cholinergic signaling pathway in lung provides a potentially important target for cholinergic drugs. This pathway may also explain some of the effects of nicotine on fetal development and also provides additional mechanisms by which smoking affects lung cancer growth and development. PMID- 14764639 TI - Improvement in sensitivity of allele-specific PCR facilitates reliable noninvasive prenatal detection of cystic fibrosis. AB - BACKGROUND: Cell-free fetal DNA circulating in maternal blood has potential as a safer alternative to invasive methods of prenatal testing for paternally inherited genetic alterations, such as cystic fibrosis (CF) mutations. METHODS: We used allele-specific PCR to detect mutated CF D1152H DNA in the presence of an excess of the corresponding wild-type sequence. Pfx buffer (Invitrogen) containing replication accessory proteins and Taq polymerase with no proofreading activity was combined with TaqMaster PCR Enhancer (Eppendorf) to suppress nonspecific amplification of the wild-type allele. The procedure was tested on DNA isolated from plasma drawn from 11 pregnant women (gestational age, 11-19.2 weeks), with mutation confirmation by chorionic villus sampling. RESULTS: The method detected 5 copies of the CF D1152H mutant allele in the presence of up to approximately 100,000 copies of wild-type allele without interference from the wild-type sequence. The D1152H mutation was correctly identified in one positive sample; the only false-positive result was seen in a mishandled sample. CONCLUSIONS: This procedure allows for reliable detection of the paternally inherited D1152H mutation and has potential application for detection of other mutations, which may help reduce the need for invasive testing. PMID- 14764640 TI - Detection of hemoglobin-based oxygen carriers in human serum for doping analysis: confirmation by size-exclusion HPLC. AB - BACKGROUND: Hemoglobin-based oxygen carriers (HBOCs) are being developed as potential substitutes for the oxygen-carrying functions of erythrocytes, but athletes may obtain and experiment with HBOCs as an illicit means of enhancing oxygen transport. An electrophoretic technique has been developed to screen for the presence of HBOCs in blood samples (Lasne et al. Clin Chem 2004;50:410-5). Interest has focused on complementary methods that can provide legally defensible scientific evidence for the presence of HBOCs in blood samples collected for doping control. METHODS: The aim of this research was to develop a size-exclusion SEC-HPLC technique to identify in plasma or serum samples the presence of HBOCs that are currently under development. This method was also used to detect a polymerized bovine hemoglobin (Hemopure) after infusion in 12 healthy males. RESULTS: The chromatograms of all HBOCs tested were clearly separated from the 54 min peak associated with human hemoglobin dimers. It was possible to differentiate between the different HBOC products based solely on their chromatographic profiles, provided they were at high concentrations. Differences were discernible not only based on the presence (or absence) of peaks, but also the separation between respective peaks. The profiles for serum samples collected from the men immediately after infusion of Hemopure showed a distinctive profile. The shape of the chromatographic profile remained consistent for at least 48 h. CONCLUSIONS: Under the analytical conditions reported here, SEC-HPLC was able to separate native hemoglobin from the modified hemoglobin molecules present in each of the HBOC products studied. In tandem with electrophoretic screening, SEC-HPLC provides evidence of the presence of HBOCs and can therefore be regarded as a method that satisfies the criteria for use in an antidoping control setting. PMID- 14764641 TI - Identification by proteomic analysis of calreticulin as a marker for bladder cancer and evaluation of the diagnostic accuracy of its detection in urine. AB - BACKGROUND: New methods for detection of bladder cancer are needed because cystoscopy is both invasive and expensive and urine cytology has low sensitivity. We screened proteins as tumor markers for bladder cancer by proteomic analysis of cancerous and healthy tissues and investigated the diagnostic accuracy of one such marker in urine. METHODS: Three specimens of bladder cancer and healthy urothelium, respectively, were used for proteome differential display using narrow-pH-range two-dimensional electrophoresis. To evaluate the presence of calreticulin (CRT) as detected by Western blotting, we obtained 22 cancerous and 10 noncancerous surgical specimens from transurethral resection or radical cystectomy. To evaluate urinary CRT, we collected 70 and 181 urine samples from patients with and without bladder cancer, respectively. Anti-CRT COOH-terminus antibody was used to detect CRT in tissue and urine. RESULTS: Proteomic analysis revealed increased CRT (55 kDa; pI 4.3) in cancer tissue. Quantitative Western blot analysis showed that CRT was increased in cancer tissue (P = 0.0003). Urinary CRT had a sensitivity of 73% (95% confidence interval, 62-83%) at a specificity of 86% (80-91%) for bladder cancer in the samples tested. CONCLUSIONS: Proteomic analysis is useful in searching for candidate proteins as biomarkers and led to the identification of urinary CRT. The diagnostic accuracy of urinary CRT for bladder cancer appears comparable to that of Food and Drug Administration-cleared urinary markers, but further studies are needed to determine its diagnostic role. PMID- 14764642 TI - Analytic performance of immunoassays for drugs of abuse below established cutoff values. AB - BACKGROUND: The analytic performance and accuracy of drug detection below Substance Abuse and Mental Health Services Administration (SAMHSA) cutoffs is not well known. In some patient populations, clinically significant concentrations of abused drugs in urine may not be detected when current SAMHSA cutoffs are used. Our objectives were to define the precision profiles of three immunoassay systems for drugs of abuse and to evaluate the accuracy of testing at concentrations at which the CV was <20%. METHODS: Drug-free urine was supplemented with analytes to assess the precision in three commercial drugs-of-abuse immunoassay systems below the SAMHSA-dictated cutoffs for amphetamines, opiates, benzoylecgonine, phencyclidine, and cannabinoids. Consecutive urine samples with signals associated with a CV <20% by Emit immunoassay and below SAMHSA cutoffs were then subjected to confirmatory analysis. RESULTS: The CV of all immunoassay systems tested remained <20% to drug concentrations well below SAMHSA cutoffs. The accuracy of urine drug-screening results between the SAMHSA-specified cutoffs and the precision-based cutoffs was less than accuracy for specimens above the SAMHSA cutoffs, but the use of the precision-based cutoff produced a 15.6% increase in the number of screen-positive specimens and a 7.8% increase in the detection of specimens that yielded positive results on confirmatory testing. CONCLUSION: The precision of three commercial immunoassay systems for drugs-of-abuse screening is adequate to detect drugs below SAMHSA cutoffs. Knowledge of the positive predictive values of screening immunoassays at lower cutoff concentrations could enable efficient use of confirmatory testing resources and improved detection of illicit drug use. PMID- 14764643 TI - Development of an immunofluorometric assay and quantification of human kallikrein 7 in tissue extracts and biological fluids. AB - BACKGROUND: Human kallikrein 7 (hK7), also known as human stratum corneum chymotryptic enzyme, is a chymotrypsin-like serine protease first identified in human skin extracts and predicted to be a secreted protease. The aim of this study was to develop a sensitive and specific immunoassay for hK7 and to examine the distribution of hK7 in tissue extracts and biological fluids. METHODS: Recombinant hK7 was produced in human embryonic kidney cells (HEK293T) and purified by a three-step column chromatographic procedure. The purified hK7 was injected into mice for antibody generation. A sandwich-type immunoassay was developed with the anti-hK7 monoclonal antibodies. RESULTS: The assay had imprecision (CV) <10% through the dynamic range of 0.2-20 microg/L and had no detectable cross-reactivity from other members in the human kallikrein gene family. Highest concentrations were found in skin, esophagus, and kidney. hK7 was also found in amniotic fluid, ascites from ovarian cancer patients, breast milk, cerebrospinal fluid, saliva, seminal plasma, serum, sweat, synovial fluid, and urine. CONCLUSIONS: This study describes the first ELISA-type immunoassay for hK7 protein quantification. hK7 is found many human tissues and in various biological fluids. PMID- 14764644 TI - Partly nonparametric approach for determining the limit of detection. AB - BACKGROUND: According to recent International Organization for Standardization (ISO) standards, the limit of detection (LoD) of an assay should be estimated taking both type I (alpha) and II (beta) errors into account. The suggested procedure, however, supposes gaussian distributions of both blank and sample measurements and a linear calibration curve. In clinical chemistry, asymmetric, nongaussian blank distributions are common, and the calibration curve may be nonlinear. We present a partly nonparametric procedure that takes these aspects into account. METHODS: Using theoretical distribution models and simulation studies, we developed a LoD estimation procedure suitable for the field of clinical chemistry that is partly based on nonparametric statistics. RESULTS: For sample size n, the nonparametrically determined 95th percentile of the blank measurements obtained as the value of the [n(95/100) + 0.5]th ordered observation defines the limit for results significantly exceeding zero [limit of blank (LoB)]. The LoD is the lowest value that is likely to yield a result exceeding the LoB. LoD is estimated as: LoB + cbeta x SDS, where SDS is the analytical SD of a sample with a low concentration; cbeta = z(1 - beta)/[1 - 1/(4 x f)]; z(1 - beta) is the standard normal deviate; and f is the number of degrees of freedom for estimation of SD(S). c(beta) is approximately equal to 1.65 for a type II error of 5%. Approaches and needed tabular values for calculation of confidence limits are presented as well as sample size. Worked examples are given to illustrate estimation and verification of the limit of detection. Simulation results are used to document performance. CONCLUSION: The proposed procedure appears useful for application in the field of clinical chemistry and promotes a standardized approach for estimating LoDs of clinical chemistry assays. PMID- 14764646 TI - Some like it plastic. PMID- 14764648 TI - Macrophage matrix metalloproteinase-9 regulates angiogenesis in ischemic muscle. PMID- 14764647 TI - Lessons from old and new kinases. PMID- 14764649 TI - Inherited arrhythmogenic diseases: the complexity beyond monogenic disorders. AB - Twelve years after the identification of the molecular bases of the long-QT syndrome, it is now possible to express some considerations on the impact that genetic findings have had in the understanding of inherited arrhythmogenic diseases. Along with the excitement for the emerging data on genotype/phenotype correlation and for the development of the first recommendations for gene specific management of patients, it is also important to acknowledge the unexpected complexity that has emerged. The focus of this article is to analyze the elusive aspects of the relationship between genetic defects and clinical manifestations and to propose some research directions that may provide the needed answers to move forward in the understanding of the genetics of heart rhythm abnormalities. PMID- 14764650 TI - Covalent and noncovalent modification of thin filament action: the essential role of troponin in cardiac muscle regulation. AB - Troponin is essential for the regulation of cardiac contraction. Troponin is a sarcomeric molecular switch, directly regulating the contractile event in concert with intracellular calcium signals. Troponin isoform switching, missense mutations, proteolytic cleavage, and posttranslational modifications are known to directly affect sarcomeric regulation. This review focuses on physiologically relevant covalent and noncovalent modifications in troponin as part of a thematic series on cardiac thin filament function in health and disease. PMID- 14764651 TI - Role of estrogens in the regulation of membrane microviscosity. PMID- 14764652 TI - Protein phosphatase 5 is a negative regulator of estrogen receptor-mediated transcription. AB - Estrogen receptors (ERs) are transcription factors that can be modulated by both estrogen-dependent and growth factor-dependent phosphorylation. A yeast two hybrid screening identified a serine/threonine protein phosphatase (PP5) as an interactant of ERbeta (1-481), a dominant negative ERbeta mutant. Glutathione S transferase pull-down assays, mammalian two-hybrid assays, and immunoprecipitation studies showed that PP5 directly binds to both ERalpha and ERbeta via its tetratricopeptide repeat domain. E domains of ERalpha and ERbeta, without containing activation domain core regions in transcription activation function 2, were required for the binding to PP5. In ERalpha-positive breast cancer MCF7 cells, estrogen- and epidermal growth factor-dependent phosphorylation of ERalpha on serine residue 118, a major phosphorylation site of the receptor, was reduced by expressing PP5 but enhanced by PP5 antisense oligonucleotide. Estrogen-induced transcriptional activities of both ERalpha and ERbeta and mRNA expression of estrogen-responsive genes, including pS2, c-myc, and cyclin D1, were suppressed by PP5 but enhanced by PP5 antisense oligonucleotide. A truncated PP5 mutant consisting only of its tetratricopeptide repeat domain acted as a dominant negative PP5 that enhanced serine residue 118 phosphorylation of ERalpha and transactivations by ERalpha and ERbeta. We present the first evidence that PP5 functions as an inhibitory regulator of ER phosphorylation and transcriptional activation in vivo. PMID- 14764653 TI - Activin regulation of the follicle-stimulating hormone beta-subunit gene involves Smads and the TALE homeodomain proteins Pbx1 and Prep1. AB - FSH is critical for normal reproductive function in both males and females. Activin, a member of the TGFbeta family of growth factors, is an important regulator of FSH expression, but little is known about the molecular mechanisms through which it acts. We used transient transfections into the immortalized gonadotrope cell line LbetaT2 to identify three regions (at -973/-962, -167, and 134) of the ovine FSH beta-subunit gene that are required for full activin response. All three regions contain homology to consensus binding sites for Smad proteins, the intracellular mediators of TGFbeta family signaling. Mutation of the distal site reduces activin responsiveness, whereas mutation of either proximal site profoundly disrupts activin regulation of the FSHbeta gene. These sites specifically bind LbetaT2 nuclear proteins in EMSAs, and the -973/-962 site binds Smad4 protein. Interestingly, the protein complex binding to the -134 site contains Smad4 in association with the homeodomain proteins Pbx1 and Prep1. Using glutathione S-transferase interaction assays, we demonstrate that Pbx1 and Prep1 interact with Smads 2 and 3 as well. The two proximal activin response elements are well conserved across species, and Pbx1 and Prep1 proteins bind to the mouse gene in vivo. Furthermore, mutation of either proximal site abrogates activin responsiveness of a mouse FSHbeta reporter gene as well, confirming their functional conservation. Our studies provide a basis for understanding activin regulation of FSHbeta gene expression and identify Pbx1 and Prep1 as Smad partners and novel mediators of activin action. PMID- 14764654 TI - Estren (4-estren-3alpha,17beta-diol) is a prohormone that regulates both androgenic and estrogenic transcriptional effects through the androgen receptor. AB - Alternative mechanisms of steroid action, through both traditional nuclear receptors and indirect pathways of gene activation, are emerging. Recent studies suggest that the synthetic steroid, 4-estrene-3alpha,17beta-diol (estren), has nongenotropic as well as sex-nonspecific osteogenic effects in ovariectomized and orchidectomized mice. We found limited estrogen receptor-dependent effects by estren on gene expression in primary osteoblast cultures and showed that it binds poorly to estrogen and androgen receptors in vitro. However, estren potently regulated direct and indirect androgen receptor-dependent effects on gene expression by osteoblasts. Consistent with this, osteoblasts produced the potent androgen 19-nortestosterone from estren by way of a 3alpha-hydroxysteroid dehydrogenase-like activity. Moreover, recombinant 3alpha-hydroxysteroid dehydrogenase (AKR1C9) and osteoblast-derived cell lysate each effectively converted estren to 19-nortestosterone in vitro, and mRNA encoding this enzyme occurs in osteoblasts. In addition to its androgenic activity, estren potently stimulated androgen receptor-dependent effects on gene expression through conventional estrogen-sensitive transcriptional elements in osteoblasts. Therefore, through local metabolism, estren indirectly activates the androgen receptor to regulate both androgen- and estrogen-like transcriptional responses by bone-forming cells. PMID- 14764655 TI - Progress and challenges in screening for early detection of ovarian cancer. AB - Ovarian cancer is characterize by few early symptoms, presentation at an advanced stage, and poor survival. As a result, it is the most frequent cause of death from gynecological cancer. During the last decade, a research effort has been directed toward improving outcomes for ovarian cancer by screening for preclinical, early stage disease using both imaging techniques and serum markers. Numerous biomarkers have shown potential in samples from clinically diagnosed ovarian cancer patients, but few have been thoroughly assessed in preclinical disease and screening. The most thoroughly investigated biomarker in ovarian cancer screening is CA125. Prospective studies have demonstrated that both CA125 and transvaginal ultrasound can detect a significant proportion of preclinical ovarian cancers, and refinements in interpretation of results have improved sensitivity and reduced the false-positive rate of screening. There is preliminary evidence that screening can improve survival, but the impact of screening on mortality from ovarian cancer is still unclear. Prospective studies of screening are in progress in both the general population and high-risk population, including the United Kingdom Collaborative Trial of Ovarian Cancer Screening (UKCTOCS), a randomized trial involving 200,000 postmenopausal women designed to document the impact of screening on mortality. Recent advances in technology for the study of the serum proteome offer exciting opportunities for the identification of novel biomarkers or patterns of markers that will have greater sensitivity and lead time for preclinical disease than CA125. Considerable interest and controversy has been generated by initial results utilizing surface-enhanced laser desorption/ionization (SELDI) in ovarian cancer. There are challenging issues related to the design of studies to evaluate SELDI and other proteomic technology, as well as the reproducibility, sensitivity, and specificity of this new technology. Large serum banks such as that assembled in UKCTOCS, which contain preclinical samples from patients who later developed ovarian cancer and other disorders, provide a unique resource for carefully designed studies of proteomic technology. There is a sound basis for optimism that further developments in serum proteomic analysis will provide powerful methods for screening in ovarian cancer and many other diseases. PMID- 14764656 TI - Activation of capsaicin-sensitive sensory neurons by carvedilol, a nonselective beta-blocker, in spontaneous hypertensive rats. AB - We performed a study in spontaneous hypertensive rats (SHR) to determine whether carvedilol, a nonselective beta-adrenoceptor antagonist, activates capsaicin sensitive sensory neurons (CSSNs), thereby promoting the release of calcitonin gene-related peptide (CGRP), a neuropeptide with an important role in maintenance of cardiovascular homeostasis. Carvedilol given intravenously at a dose of 0.3 mg/kg transiently decreased the mean arterial blood pressure (MABP) and increased renal tissue blood flow with increases in CGRP levels in plasma and kidney. These effects induced by carvedilol were not seen in animals pretreated with capsazepine, an antagonist of capsaicin. Although 1.0 mg/kg cavedilol markedly decreased MABP, it neither increased renal tissue blood flow nor CGRP levels in plasma and kidney. Prazosin, a selective alpha(1)-adrenoceptor antagonist, and bisoprolol, a selective beta(1)-adrenoceptor antagonist, decreased MABP with capsazepine, showing no antagonistic action in either cases, and these agents increased neither renal tissue blood flow nor levels of CGRP in plasma and kidney. Both ICI 118,551 [(+/-)-1-[2,3-(dihydro-7-methyl-1H-inden-4-yl)oxy]-3-[(1 methylethyl)amino]-2-butanol], a selective beta(2)-adrenoceptor antagonist, at a dose of 0.25 mg/kg and capsaicin mimicked effects induced by 0.3 mg/kg carvedilol. Administration of 1.0 mg/kg ICI 118,551 produced effects similar to those induced by 1.0 mg/kg carvedilol. These observations strongly suggested that the low dose of carvedilol might activate CSSNs in SHR to increase the release of CGRP, thereby decreasing blood pressure with an increase in renal tissue blood flow. The effects induced by carvedilol seemed to be mediated by its beta(2) adrenoceptor blockade activity. PMID- 14764657 TI - Anthraquinone polyamines: novel channel blockers to study N-methyl-D-aspartate receptors. AB - The effects of various anthraquinone polyamines (AQP) were studied at recombinant N-methyl-d-aspartate (NMDA) receptors expressed in Xenopus laevis oocytes. The AQP derivatives had different numbers of methylene groups between the NH(2) (or NH) groups in their spermidine-like tail. Thus, we termed these derivatives AQ33, AQ34, etc. All AQP derivatives inhibited responses of NR1/NR2 receptors in oocytes voltage-clamped at -70 mV, with IC(50) values between 4 and 22 microM. The block was strongly voltage-dependent. AQ34 and AQ33b inhibited responses of NR1/NR2 receptors but did not inhibit responses of alpha-amino-3-hydroxy-5-methyl 4-isoxazolepropionic acid (AMPA) receptors expressed from GluR1 or GluR2(Q), indicating that AQ34 and AQ33b are preferential NMDA antagonists. Results of experiments using mutant NR1 and NR2 subunits identified residues that influence block by AQ34 and AQ33b. These residues are located in the outer vestibule at the selectivity filter/narrowest constriction of the channel and in the inner vestibule below the level of the selectivity filter. The results with mutant NR1 and NR2 subunits are consistent with the idea that NR1(Asn616) and NR2B(Asn616), but not NR2B(Asn615), make the narrowest constriction of NMDA channel. PMID- 14764658 TI - Potassium canrenoate, an aldosterone receptor antagonist, reduces isoprenaline induced cardiac fibrosis in the rat. AB - The purpose of the present study was to determine whether the administration of an antagonist of aldosterone could prevent the fibrosis induced by an acute injection of isoprenaline. Male Wistar rats were submitted to one subcutaneous injection of isoprenaline (400 mg/kg) and were simultaneously treated with potassium canrenoate in drinking water (20 mg/kg/day) started 5 days before the injection of isoprenaline. Two months later, echocardiographic and hemodynamic measurements were performed. Then, the heart was prepared for morphometric histology and quantification of fibrosis in the left ventricle. Heart and left ventricular weights were increased significantly by isoprenaline. Potassium canrenoate attenuated this increase. The administration of isoprenaline increased significantly end diastolic diameter and end systolic volume compared with control. These changes were increased further with the addition of potassium canrenoate. In contrast, the fibrosis induced by isoprenaline was reduced significantly by potassium canrenoate at the three section levels. Potassium canrenoate attenuated the fibrosis but not the enhanced dilatation of the left ventricle induced by isoprenaline. PMID- 14764659 TI - Deletion of the fyn-kinase gene alters sensitivity to GABAergic drugs: dependence on beta2/beta3 GABAA receptor subunits. AB - Tyrosine phosphorylation can modulate GABA(A) receptor function, and deletion of the fyn-kinase gene alters GABAergic function in olfactory bulb neurons, as reported by Kitazawa, Yagi, Miyakawa, Niki, and Kawai (J Neurophysiol 1998;79:137 142). Our goal was to determine whether fyn gene deletion altered behavioral and functional actions of compounds that act on GABA(A) receptors. Such evidence might suggest a role for fyn-kinase in modulating GABA(A) receptor function, possibly via direct interactions between the kinase and receptor. Using the loss of righting reflex test, we found that null mutants were less sensitive to the hypnotic effects of THIP (4,5,6,7-tetrahydroisoxazolo[5,4-c]pyridin-3-ol), a GABA(A) receptor agonist. Subunit specificity was suggested by the observation that null mutants were also less sensitive to the hypnotic effects of etomidate, a GABAergic compound that is selective for receptors possessing beta2 and/or beta3 receptor subunits. The genotypes did not differ in sensitivity to zolpidem, an alpha1-selective GABAergic drug. GABA(A) receptor functional assays ((36)Cl(-) influx) supported our behavioral results; the actions of the GABA(A) agonists, THIP and muscimol, were reduced in the cerebellar membranes of fyn-null mutant mice. Importantly, similar results were seen with etomidate. Binding of [(3)H]flunitrazepam supported the idea that this is due to a decrease in functional GABA(A) receptor density. These data suggest that fyn-kinase may alter the function of GABA(A) receptors, perhaps via actions on beta2 and/or beta3 receptor subunits. PMID- 14764660 TI - Cutting edge: in vitro-generated tolerogenic APC induce CD8+ T regulatory cells that can suppress ongoing experimental autoimmune encephalomyelitis. AB - APC exposed to TGFbeta2 and Ag (tolerogenic APC) promote peripheral Ag-specific tolerance via the induction of CD8(+) T regulatory cells capable of suppressing Th1 and Th2 immunity. We postulated that tolerogenic APC might reinstate tolerance toward self-neuronal Ags and ameliorate ongoing experimental autoimmune encephalomyelitis (EAE). Seven days after immunization with myelin basic protein (MBP), mice received MBP-specific tolerogenic APC, and EAE was evaluated clinically. To test for the presence and the phenotype of T regulatory cells, CD4 and/or CD8 T cells from tolerogenic APC-treated mice were transferred to naive mice before their immunization with MBP. The MBP-specific tolerogenic APC decreased both the severity and incidence of ongoing EAE. Tolerance to self neuronal Ags was induced in naive recipient mice via adoptive transfer of CD8(+), but not CD4(+) T cells. Rational use of in vitro-generated tolerogenic APC may lead to novel therapy for autoimmune disease. PMID- 14764661 TI - Cutting edge: conventional CD8 alpha+ dendritic cells are generally involved in priming CTL immunity to viruses. AB - Dendritic cells (DCs) play a central role in initiating immune responses. Despite this, there is little understanding how different DC subsets contribute to immunity to different pathogens. CD8alpha(+) DC have been shown to prime immunity to HSV. Whether this very limited capacity of a single DC subset priming CTL immunity is restricted to HSV infection or is a more general property of anti viral immunity was examined. Here, we show that the CD8alpha(+) DCs are the principal DC subset that initiates CTL immunity to s.c. infection by influenza virus, HSV, and vaccinia virus. This same subset also dominated immunity after i.v. infection with all three viruses, suggesting a similar involvement in other routes of infection. These data highlight the general role played by CD8alpha(+) DCs in CTL priming to viral infection and raises the possibility that this DC subset is specialized for viral immunity. PMID- 14764662 TI - Cutting edge: Toll-like receptor signaling in macrophages induces ligands for the NKG2D receptor. AB - Macrophages recognize the presence of infection by using the Toll-like receptor (TLR) family of proteins that detect ligands on bacterial, viral, and fungal pathogens. We show that murine macrophages stimulated with pathogen products known to signal through TLRs express ligands for the NKG2D receptor, found on NK cells, activated CD8(+) T cells and activated macrophages. TLR signaling, through the MyD88 adaptor, up-regulates transcription of the retinoic acid early inducible-1 (RAE-1) family of NKG2D ligands, but not H-60 or murine UL16-binding protein-like transcript-1. RAE-1 proteins are found on the surface of activated, but not resting, macrophages and can be detected by NKG2D on NK cells resulting in down-regulation of this receptor both in vitro and in vivo. RAE-1-NKG2D interactions provide a mechanism by which NK cells and infected macrophages communicate directly during an innate immune response to infection. PMID- 14764663 TI - Cutting edge: IL-26 signals through a novel receptor complex composed of IL-20 receptor 1 and IL-10 receptor 2. AB - The receptor for IL-26 (AK155), a cytokine of the IL-10 family, has not previously been defined. We demonstrate that the active receptor complex for IL 26 is a heterodimer composed of two receptor proteins: IL-20R1 and IL-10R2. Signaling through the IL-26R results in activation of STAT1 and STAT3 which can be blocked by neutralizing Abs against IL-20R1 or IL-10R2. IL-10R2 is broadly expressed on a wide variety of tissues, whereas only a limited number of tissues express IL-20R1. Therefore, the ability to respond to IL-26 is restricted by the expression of IL-20R1. IL-10, IL-19, IL-20, IL-22, and IL-24 fail to signal through the combination of IL-10R2 and IL-20R1 proteins, demonstrating that this receptor combination is unique and specific for IL-26. PMID- 14764664 TI - Cutting edge: T cells trigger CD40-dependent platelet activation and granular RANTES release: a novel pathway for immune response amplification. AB - Platelets, in addition to exerting hemostatic activity, contribute to immunity and inflammation. The recent report that platelets express CD40 led us to hypothesize that CD40 ligand (CD40L)-positive T cells could bind to platelets, cause their activation, and trigger granular RANTES release, creating a T cell recruitment feedback loop. Platelets were cocultured with resting or activated autologous T cells and their activation was assessed by P-selectin expression. RANTES binding to endothelial cells was assessed by confocal microscopy, and its biological activity was demonstrated by a T cell adhesion assay. CD40L-positive T cells induced platelet activation through a contact-mediated, CD40-dependent pathway resulting in RANTES release, which bound to endothelial cells and mediated T cell recruitment. Soluble CD40L induced the same events via p38, but not extracellular signal-regulated kinase, phosphorylation. These results show the existence of a novel platelet-dependent pathway of immune response amplification which brings these nonimmune cells close to the level of pathogenic relevance traditionally attributed to classical immune cells. PMID- 14764665 TI - Cutting edge: dendritic cells copulsed with microbial and helminth antigens undergo modified maturation, segregate the antigens to distinct intracellular compartments, and concurrently induce microbe-specific Th1 and helminth-specific Th2 responses. AB - To examine the ability of dendritic cells (DC) to discriminate between helminth and microbial Ag and induce appropriately polarized Th responses, mouse DC were copulsed with the helminth Ag, schistosome egg Ag (SEA), along with the bacterium Proprionebacterium acnes, Pa, and transferred into wild-type mice. Strikingly, SEA/Pa-copulsed DC induced concurrent Pa-specific Th1 (but not Th2) responses and SEA-specific Th2 (but not Th1) responses. Although DC exposed to both Ag undergo many of the maturation-associated changes that accompany exposure to Pa alone, Pa induced IL-12 production was inhibited by SEA. Examination of Ag uptake revealed that SEA and Pa are acquired via discrete pathways and enter nonoverlapping intracellular compartments. Data suggest that segregation of SEA and Pa into distinct compartments, coupled with SEA-induced modifications of the DC maturation pathway, are significant components of the ability of DC to interpret signals inherent to SEA and Pa and induce appropriately polarized Th responses. PMID- 14764666 TI - Visualizing the site and dynamics of IgG salvage by the MHC class I-related receptor, FcRn. AB - The MHC class I-related receptor, FcRn, plays a central role in regulating the serum levels of IgG. FcRn is expressed in endothelial cells, suggesting that these cells may be involved in maintaining IgG levels. We have used live cell imaging of FcRn-green fluorescent protein transfected human endothelial cells to analyze the intracellular events that control IgG homeostasis. We show that segregation of FcRn-IgG complexes from unbound IgG occurs in the sorting endosome. FcRn or FcRn-IgG complexes are gradually depleted from sorting endosomes to ultimately generate multivesicular bodies whose contents are destined for lysosomal degradation. In addition, the pathways taken by FcRn and the transferrin receptor overlap, despite distinct mechanisms of ligand uptake. The studies provide a dynamic view of the trafficking of FcRn and its ligand and have relevance to understanding how FcRn functions to maintain IgG homeostasis. PMID- 14764667 TI - Visualization of antigen presentation by actin-mediated targeting of glycolipid enriched membrane domains to the immune synapse of B cell APCs. AB - Glycolipid-enriched membrane (GEM) domains, or lipid rafts, function in signaling in immune cells, but their properties during Ag presentation are less clear. To address this question, GEM domains were studied using fluorescence cell imaging of mouse CH27 B cells presenting Ag to D10 T cells. Our experiments showed that APCs were enriched with GEM domains in the immune synapse, and this occurred in an actin-dependent manner. This enrichment was specific to GEM domains, because a marker for non-GEM regions of the membrane was excluded from the immune synapse. Furthermore, fluorescence photobleaching experiments showed that protein in the immune synapse was dynamic and rapidly exchanged with that in other compartments of CH27 cells. To identify the signals for targeting GEM domains to the immune synapse in APCs, capping of the domains was measured in cells after cross-linking surface molecules. This showed that co-cross-linking CD48 with MHC class II was required for efficient capping and intracellular signaling. Capping of GEM domains by co-cross-linking CD48 and MHC class II occurred with co-capping of filamentous actin, and both domain capping and T cell-CH27 cell conjugation were inhibited by pretreating CH27 cells with latrunculin B. Furthermore, disruption of the actin cytoskeleton of the CH27 cells also inhibited formation of a mature immune synapse in those T cells that did conjugate to APCs. Thus, Ag presentation and efficient T cell stimulation occur by an actin-dependent targeting of GEM domains in the APC to the site of T cell engagement. PMID- 14764668 TI - Inhibitory effect of Toll-like receptor 4 on fusion between phagosomes and endosomes/lysosomes in macrophages. AB - Toll-like receptor 4 (TLR4) of macrophages recognizes LPS of Gram-negative bacteria in cooperation with CD14, which is also involved in the recognition of apoptotic cells. In this study we asked whether TLR4 plays a role in the phagocytic clearance of apoptotic cells by macrophages. Macrophages were prepared from peritoneal fluid of thioglycolate-treated mice carrying either a wild-type or a disrupted TLR4-encoding gene and were examined for their ability to phagocytose apoptotic mouse thymocytes, apoptotic Jurkat T cells, Ig-opsonized mouse thymocytes, Ig-opsonized zymosan particles, and latex beads. Both populations of macrophages equally expressed CD14 on their surfaces and showed almost equal activities of binding to and engulfing all these targets. However, apoptotic thymocytes, apoptotic Jurkat cells, and opsonized thymocytes disappeared more rapidly in TLR4-deficient macrophages than in wild-type macrophages, and the fusion between endosomes/lysosomes and phagosomes containing any target cells or particles was accelerated in mutant macrophages. Activation of the transcription factor NF-kappaB appeared not to occur in wild-type macrophages after engulfment, and the rate of apoptotic cell degradation in wild type macrophages remained the same regardless of the activation of NF-kappaB. Finally, immunohistochemical analyses showed that ectopically expressed TLR4 was associated with phagosomes in a macrophage-derived cell line. All these results collectively indicate that TLR4 negatively regulates the degradation of engulfed cells in macrophages via a pathway independent of NF-kappaB. PMID- 14764669 TI - IL-21 induces the functional maturation of murine NK cells. AB - IL-21 is a recently identified cytokine that stimulates mouse NK cell effector functions in vitro. In this study we demonstrate that IL-21 achieves its stimulatory effect by inducing the development of mature NK cells into a large granular lymphocyte phenotype with heightened effector function. IL-21 treatment results in increased cell size and granularity and a corresponding decrease in cell viability and proliferative potential. These cells up-regulate the expression of the inhibitory CD94-NKG2A receptor complex and the activation markers CD154 and killer cell, lectin-like-receptor G1. Surprisingly, IL-21 treatment also results in down-regulation of the pan-NK marker, NK1.1. Coinciding with these cellular changes IL-21 enhances cytolytic capacity across a spectrum of target sensitivities and induces IL-10 and IFN-gamma production. In vivo treatment with IL-21 results in a very similar activation and phenotypic maturation of NK cells as well as a potent increase in NK cell-mediated anti tumor immunity that is perforin dependent. These developmental changes suggested that IL-21 functions to induce the terminal differentiation of mouse NK cells, resulting in heightened NK cell-mediated cytotoxicity and immune surveillance. PMID- 14764670 TI - IL-4 induces differentiation and expansion of Th2 cytokine-producing eosinophils. AB - Innate effector cells that produce Th2-type cytokines are critical in Th2 cell mediated immune responses. However, it is not known how these cells acquire the ability to produce Th2 cytokines. IL-4 is a potent inducer that directs differentiation of naive CD4(+) T cells into CD4(+) Th2 effector cells. To determine whether IL-4 can induce differentiation and expansion of Th2 cytokine producing innate cells, we used mice whose il-4 gene was replaced by a knock-in green fluorescence protein (gfp) gene. We found that, directly ex vivo, IL-4 increased the number of GFP(+) cells in the airway and the lung tissue in an Ag specific manner. The majority of GFP(+) cells were eosinophils, suggesting that IL-4 plays a pivotal role in expanding IL-4-producing eosinophils in vivo. IL-4 producing eosinophils showed some unique features compared with IL-4-producing CD4(+) T cells. They exhibited biallelic expression of the il-4 gene when stimulated and were more dominant IL-4- and IL-5-producing cells. Furthermore, we show that IL-4 drove bone marrow progenitor cells to differentiate into Th2 cytokine-producing eosinophils in vitro. These results strongly suggest IL-4 is a potent factor in directing bone marrow progenitor cells to differentiate into Th2 cytokine-producing eosinophils. PMID- 14764671 TI - NF-kappa B-inducing kinase establishes self-tolerance in a thymic stroma dependent manner. AB - Physical contact between thymocytes and the thymic stroma is essential for T cell maturation and shapes the T cell repertoire in the periphery. Stromal elements that control these processes still remain elusive. We used a mouse strain with mutant NF-kappaB-inducing kinase (NIK) to examine the mechanisms underlying the breakdown of self-tolerance. This NIK-mutant strain manifests autoimmunity and disorganized thymic structure with abnormal expression of Rel proteins in the stroma. Production of immunoregulatory T cells that control autoreactive T cells was impaired in NIK-mutant mice. The autoimmune disease seen in NIK-mutant mice was reproduced in athymic nude mice by grafting embryonic thymus from NIK-mutant mice, and this was rescued by supply of exogenous immunoregulatory T cells. Impaired production of immunoregulatory T cells by thymic stroma without normal NIK was associated with altered expression of peripheral tissue-restricted Ags, suggesting an essential role of NIK in the thymic microenvironment in the establishment of central tolerance. PMID- 14764672 TI - Mechanisms governing B cell developmental defects in invariant chain-deficient mice. AB - Invariant chain (Ii)-deficient mice exhibit profound B cell defects that have remained poorly understood, because they could not be simply explained by impaired Ag presentation. We found that Ii deficiency induced cell autonomous defects of two distinct B cell lineages. The life span of mature follicular (FO) B cells was reduced, accounting for their markedly decreased frequency, whereas, in contrast, marginal zone (MZ) B cells accumulated. Other Ii-expressing lineages such as B1 B cells and dendritic cells were unaffected. Surprisingly, the life span of FO B cells was fully corrected in Ii/I-Abeta doubly deficient mice, revealing that Ii-free I-Abeta chains alter FO B cell survival. In contrast, the accumulation of MZ B cells was controlled by a separate mechanism independent of I-Abeta. Interestingly, in Ii-deficient mice lacking FO B cells, the MZ B cells invaded the FO zone, suggesting that intact follicules contribute to the retention of B cells in the MZ. These findings reveal unexpected consequences of Ii deficiency on the development and organization of B cell follicles. PMID- 14764673 TI - B cell receptor-mediated apoptosis of human lymphocytes is associated with a new regulatory pathway of Bim isoform expression. AB - Studies in Bim-deficient mice have shown that the proapoptotic molecule Bim plays a key role in the control of B cell homeostasis and activation. However, the role of Bim in human B lymphocyte apoptosis is unknown. We show in this study that, depending on the degree of cross-linking, B cell receptors can mediate both Bim dependent and apparent Bim-independent apoptotic pathways. Cross-linked anti-mu Ab-mediated activation induces an original pathway governing the expression of the various Bim isoforms. This new pathway involves the following three sequential steps: 1) extracellular signal-regulated kinase-dependent phosphorylation of the BimEL isoform, which is produced in large amounts in healthy B cells; 2) proteasome-mediated degradation of phosphorylated BimEL; and 3) increased expression of the shorter apoptotic isoforms BimL and BimS. PMID- 14764674 TI - Transgenic expression of a human polyreactive Ig expressed in chronic lymphocytic leukemia generates memory-type B cells that respond to nonspecific immune activation. AB - We generated transgenic mice, designated SMI, expressing unmutated H and L chain Ig genes encoding a low-affinity, polyreactive human (h)IgM/kappa rheumatoid factor. These animals were compared with control AB29 transgenic mice expressing a hIgM/kappa rheumatoid factor specific for human IgG, with no detectable reactivity with mouse proteins. SMI B cells expressed significantly lower levels of surface hIgM/kappa than did the B cells of AB29 mice, but still could be induced to proliferate by surface Ig cross-linking in vitro and could be deleted with anti-Id mAb in vivo. Transgene-expressing B cells of AB29 mice had a B-2 phenotype and were located in the primary follicle. In contrast, a relatively high proportion of hIgM-expressing B cells of SMI mice had the phenotype of B-1 B cells in the peritoneum or marginal zone B cells in the spleen, where they were located in the periarteriolar sheath, marginal zone, and interfollicular areas that typically are populated by memory-type B cells. Although the relative proportions of transgene-expressing B cells in both types of transgenic mice declined with aging, SMI mice experienced progressive increases in the serum levels of IgM transgene protein over time. Finally, SMI transgene-expressing B cells, but not AB29 transgene-expressing B cells, were induced to secrete Ab when cultured with alloreactive T cells. These results indicate that expression of polyreactive autoantibodies can allow for development of B cells that are neither deleted nor rendered anergic, but instead have a phenotype of memory-type or Ag experienced B cells that respond to nonspecific immune activation. PMID- 14764676 TI - Distinct expression profiles of the peripheral cannabinoid receptor in lymphoid tissues depending on receptor activation status. AB - Using two distinct anti-CB2 receptor Abs, we investigated the expression patterns of the peripheral cannabinoid receptor CB2 in human secondary lymphoid organs. Immunohistochemical analysis using an N-terminal specific anti-CB2 Ab revealed high protein expression in the germinal centers (GCs) of secondary follicles. A C terminal specific anti-CB2 Ab, which only recognizes a nonphosphorylated inactive receptor, showed positivity in the mantle zones (MZs) and marginal zones (MGZs) of the secondary follicles where resting cells reside, and in the primary follicles. In contrast, no positivity was observed in GCs using the C-terminal Ab, suggesting that active CB2 receptors are mainly present on cells in the GCs. Dual immunohistochemical analysis revealed that B lymphocytes express the CB2 protein abundantly. In contrast to B cells in the MZ or MGZ, CB2-expressing cells in the GCs coexpress the costimulatory membrane protein CD40, which is mainly expressed in the GCs and at very low levels in the MZs and MGZs and the proliferation marker Ki-67. Using the human Raji B cell line as a model, we demonstrate in a transwell assay that moderate migration occurs upon stimulation of the CB2 receptor with the endocannabinoid 2-arachidonoylglycerol, which is enhanced by CD40 costimulation. Our findings, that GC-related cells express active CB2 and that CB2-dependent migration requires CD40 costimulation, suggest that CB2 is involved in B cell activation. PMID- 14764675 TI - Severely impaired B lymphocyte proliferation, survival, and induction of the c Myc:Cullin 1 ubiquitin ligase pathway resulting from CD22 deficiency on the C57BL/6 genetic background. AB - Understanding the molecular mechanisms through which CD22 regulates B lymphocyte homeostasis, signal transduction, and tolerance is critical to defining normal B cell function and understanding the role of CD22 in autoimmunity. Therefore, CD22 function was examined in vivo and in vitro using B cells from CD22-deficient (CD22(-/-)) mice. Backcrossing of founder CD22(-/-) mice onto the C57BL/6 (B6) genetic background from a B6/129 mixed background resulted in a dramatically reduced B cell proliferative response following IgM ligation, characterized by a paucity of lymphoblasts and augmented apoptosis. Also, the phenotype of splenic B6 CD22(-/-) B cells was uniquely HSA(high) and IgD(low)/CD21(low) with intermediate levels of CD5 expression, although the percentages of mature and transitional B cells were normal. That B6 CD22(-/-) B cells predominantly underwent apoptosis following IgM ligation correlated with this unique tolerant phenotype, as well as defective induction of the c-Myc:Cullin 1 (CUL1) ubiquitin ligase pathway that is necessary for progression to the S phase of cell cycle. CD40 ligation compensated for CD22 deficiency by restoring lymphoblast development, proliferation, c-Myc and CUL1 expression, and protein ubiquitination/degradation in IgM-stimulated B6 CD22(-/-) B cell cultures. Thereby, this study expands our current understanding of the complex role of CD22 during B cell homeostasis and Ag responsiveness, and reveals that the impact of CD22 deficiency is dictated by the genetic background on which it is rendered. Moreover, this study defines CD22 and CD40 as the first examples of lymphocyte coreceptors that influence induction of the c-Myc:CUL1 ubiquitin ligase pathway. PMID- 14764677 TI - Complete loss of Fas ligand gene causes massive lymphoproliferation and early death, indicating a residual activity of gld allele. AB - To investigate the in vivo function of Fas ligand (FasL), we produced a mouse strain with a FasL gene flanked by loxP sequences. Mice with homozygous floxed FasL gene showed no obvious abnormalities. However, germline deletion of the FasL gene, obtained after mating with mice expressing ubiquitous Cre recombinase, resulted in an unexpectedly severe phenotype. FasL(-/-) mice exhibited an extreme splenomegaly and lymphadenopathy associated with lymphocytic infiltration into multiple organs and autoimmune disease. This severe phenotype led to the premature death at 4 mo of age of >50% of the homozygous mice. It stands in sharp contrast with the milder disease observed in gld (generalized lymphoproliferative disease) mice, indicating that the FasL allele of these mice encodes a protein still able to bind, albeit at a very low level, the Fas receptor. PMID- 14764678 TI - Exosomes as potent cell-free peptide-based vaccine. I. Dendritic cell-derived exosomes transfer functional MHC class I/peptide complexes to dendritic cells. AB - Current immunization protocols in cancer patients involve CTL-defined tumor peptides. Mature dendritic cells (DC) are the most potent APCs for the priming of naive CD8(+) T cells, eventually leading to tumor eradication. Because DC can secrete MHC class I-bearing exosomes, we addressed whether exosomes pulsed with synthetic peptides could subserve the DC function consisting in MHC class I restricted, peptide-specific CTL priming in vitro and in vivo. The priming of CTL restricted by HLA-A2 molecules and specific for melanoma peptides was performed: 1) using in vitro stimulations of total blood lymphocytes with autologous DC pulsed with GMP-manufactured autologous exosomes in a series of normal volunteers; 2) in HLA-A2 transgenic mice (HHD2) using exosomes harboring functional HLA-A2/Mart1 peptide complexes. In this study, we show that: 1). DC release abundant MHC class I/peptide complexes transferred within exosomes to other naive DC for efficient CD8(+) T cell priming in vitro; 2). exosomes require nature's adjuvants (mature DC) to efficiently promote the differentiation of melanoma-specific effector T lymphocytes producing IFN-gamma (Tc1) effector lymphocytes in HLA-A2 transgenic mice (HHD2). These data imply that exosomes might be a transfer mechanism of functional MHC class I/peptide complexes to DC for efficient CTL activation in vivo. PMID- 14764679 TI - Exosomes as potent cell-free peptide-based vaccine. II. Exosomes in CpG adjuvants efficiently prime naive Tc1 lymphocytes leading to tumor rejection. AB - Ideal vaccines should be stable, safe, molecularly defined, and out-of-shelf reagents efficient at triggering effector and memory Ag-specific T cell-based immune responses. Dendritic cell-derived exosomes could be considered as novel peptide-based vaccines because exosomes harbor a discrete set of proteins, bear functional MHC class I and II molecules that can be loaded with synthetic peptides of choice, and are stable reagents that were safely used in pioneering phase I studies. However, we showed in part I that exosomes are efficient to promote primary MHC class I-restricted effector CD8(+) T cell responses only when transferred onto mature DC in vivo. In this work, we bring evidence that among the clinically available reagents, Toll-like receptor 3 and 9 ligands are elective adjuvants capable of triggering efficient MHC-restricted CD8(+) T cell responses when combined to exosomes. Exosome immunogenicity across species allowed to verify the efficacy of good manufactory procedures-manufactured human exosomes admixed with CpG oligonucleotides in prophylactic and therapeutic settings of melanoma in HLA-A2 transgenic mice. CpG adjuvants appear to be ideal adjuvants for exosome-based cancer vaccines. PMID- 14764680 TI - Novel regulatory mechanisms of CD40-induced prostanoid synthesis by IL-4 and IL 10 in human monocytes. AB - Interleukins IL-4 and IL-10 are considered to be central regulators for the limitation and eventual termination of inflammatory responses in vivo, based on their potent anti-inflammatory effects toward LPS-stimulated monocytes/macrophages and neutrophils. However, their role in T cell-dependent inflammatory responses has not been fully elucidated. In this study, we investigated the effects of both cytokines on the production of PGE(2), a key molecule of various inflammatory conditions, in CD40-stimulated human peripheral blood monocytes. CD40 ligation of monocytes induced the synthesis of a significant amount of PGE(2) via inducible expression of the cyclooxygenase (COX) 2 gene. Both IL-10 and IL-4 significantly inhibited PGE(2) production and COX-2 expression in CD40-stimulated monocytes. Using specific inhibitors for extracellular signal-related kinase (ERK) and p38 mitogen-activated protein kinase (MAPK), we found that both kinase pathways are involved in CD40-induced COX-2 expression. CD40 ligation also resulted in the activation of NF-kappaB. Additional experiments exhibited that CD40 clearly induced the activation of the upstream kinases MAPK/ERK kinase 1/2, MAPK kinase 3/6, and I-kappaB in monocytes. IL-10 significantly inhibited CD40-induced activation of the ERK, p38 MAPK, and NF-kappaB pathways; however, inhibition by IL-4 was limited to the ERK pathway in monocytes. Neither IL-10 nor IL-4 affected the recruitment of TNFR-associated factors 2 and 3 to CD40 in monocytes. Collectively, IL-10 and IL-4 use novel regulatory mechanisms for CD40-induced prostanoid synthesis in monocytes, thus suggesting a potential role for these cytokines in regulating T cell-induced inflammatory responses, including autoimmune diseases. PMID- 14764681 TI - Reduced Ig class switch in aged mice correlates with decreased E47 and activation induced cytidine deaminase. AB - The capacity to class switch the IgH chain is critical to the effectiveness of humoral immune responses. We show that in vitro-stimulated splenic B cells from senescent mice are deficient in production of multiple class switch isotypes (IgG1, G2a, G3, and E), class switch recombination (CSR), and induction of the E2A-encoded transcription factor E47. E47 has previously been shown to be required for CSR, at least in part via expression of the activation-induced cytidine deaminase. Our studies show that impaired induction of E47, and subsequently activation-induced cytidine deaminase, contribute to poor CSR and production of secondary isotypes in senescence. PMID- 14764682 TI - Dependency of direct pathway CD4+ T cells on CD40-CD154 costimulation is determined by nature and microenvironment of primary contact with alloantigen. AB - Blockade of the CD40-CD154 costimulatory pathway can inhibit CD4(+) T cell mediated alloimmune responses. The aim of this study was to define the in vivo requirement for CD40-CD154 costimulation by CD4(+) T cells that respond to alloantigen following direct recognition. We used TCR-transgenic CD4(+) T cells that are reactive to the MHC class II alloantigen, H2A(s). An experimental in vivo model was established that allowed direct comparison of the fate of a trace population of H2A(s)-reactive CD4(+) T cells when challenged with different forms of H2A(s+) alloantigen under conditions of CD40-CD154 costimulation blockade. In this study, we demonstrate that an i.v. infusion of H2A(s+) leukocytes in combination with anti-CD154 therapy rapidly deletes H2A(s)-reactive CD4(+) T cells. In contrast, following transplantation of an H2A(s+) cardiac allograft, H2A(s)-reactive CD4(+) T cell responses were unaffected by blocking CD40-CD154 interactions. Consistent with these findings, combined treatment with donor leukocytes and anti-CD154 therapy was found to be more effective in prolonging the survival of cardiac allografts compared with CD154 mAb treatment alone. The dominant mechanism by which donor leukocyte infusion and anti-CD154 therapy facilitate allograft acceptance is deletion of donor-reactive direct pathway T cells. No evidence for the generation of regulatory cells by this combined therapy was found. Taken together, these results clearly demonstrate that naive alloreactive CD4(+) T cells have distinct requirements for CD40-CD154 costimulation depending on the form and microenvironment of primary alloantigen contact. PMID- 14764683 TI - Disparate in vitro and in vivo requirements for IL-2 during antigen-independent CD8 T cell expansion. AB - Transient TCR stimulation induces multiple rounds of CD8 T cell division without further requirement for Ag. The mechanism driving Ag-independent proliferation, however, remains unclear. In this study, we show that the initial duration of TCR stimulation positively correlates with the number of divisions that CD8 T cells subsequently undergo. We find that increased periods of Ag stimulation result in enhanced CD25 up-regulation and greater IL-2 production by CD8 T cells. Depletion of IL-2 from T cell cultures with specific Abs dramatically impairs programmed proliferation. Consistent with this result, IL-2-deficient T cells undergo markedly attenuated Ag-independent proliferation in vitro. Although IL-2 production by stimulated CD8 T cells appears to be essential for in vitro proliferation, upon transfer into recipient mice, IL-2-deficient CD8 T cells undergo extensive proliferation in vivo after transient stimulation. Furthermore, the extent of in vivo proliferation correlates with the duration of in vitro Ag stimulation. These results indicate that the requirements for autocrine IL-2 production by CD8 T cells differs between in vitro and in vivo conditions and suggests that factors in addition to IL-2 can support Ag-independent CD8 T cell proliferation. PMID- 14764684 TI - A novel role for an insect apolipoprotein (apolipophorin III) in beta-1,3-glucan pattern recognition and cellular encapsulation reactions. AB - Lipoproteins and molecules for pattern recognition are centrally important in the innate immune response of both vertebrates and invertebrates. Mammalian apolipoproteins such as apolipoprotein E (apoE) are involved in LPS detoxification, phagocytosis, and possibly pattern recognition. The multifunctional insect protein, apolipophorin III (apoLp-III), is homologous to apoE. In this study we describe novel roles for apoLp-III in pattern recognition and multicellular encapsulation reactions in the innate immune response, which may be of direct relevance to mammalian systems. It is known that apoLp-III stimulates antimicrobial peptide production in insect blood, enhances phagocytosis by insect blood cells (hemocytes), and binds and detoxifies LPS and lipoteichoic acid. In the present study we show that apoLp-III from the greater wax moth, Galleria mellonella, also binds to fungal conidia and beta-1,3-glucan and therefore may act as a pattern recognition molecule for multiple microbial and parasitic invaders. This protein also stimulates increases in cellular encapsulation of nonself particles by the blood cells and exerts shorter term, time-dependent, modulatory effects on cell attachment and spreading. All these responses are dose dependent, occur within physiological levels, and, with the notable exception of beta-glucan binding, are only observed with the lipid associated form of apoLp-III. Preliminary studies also established a beneficial role for apoLp-III in the in vivo response to an entomopathogenic fungus. These data suggest a wide range of immune functions for a multiple specificity pattern recognition molecule and may provide a useful model for identifying further potential roles for homologous proteins in mammalian immunology, particularly in terms of fungal infections, pneumoconiosis, and granulomatous reactions. PMID- 14764685 TI - Extremely rapid and intense induction of apoptosis in human eosinophils by anti CD30 antibody treatment in vitro. AB - Apoptosis is an important cellular mechanism for controlling cell viability and proliferation. With respect to eosinophils, cytokines prolong their survival, whereas corticosteroids reduce their survival in vitro. CD30, a member of the TNFR family, is expressed on the surface of many cell types, including Hodgkin's lymphoma cells. CD30 is capable of inducing apoptosis after Ab treatment in some cell lines. To determine whether this surface structure is involved in apoptosis of human eosinophils, we examined its expression and the effect of anti-CD30 Ab treatment on the viability of eosinophils. Purified human eosinophils expressed low, but consistently detectable, levels of CD30. Immobilized, but not soluble, forms of anti-CD30 Abs (HRS-4 and Ber-H8) or recombinant mouse CD30 ligand exhibited an extremely rapid and intense survival-reducing effect on the eosinophils in the presence of exogenous IL-5; this effect was both concentration and time dependent. Furthermore, high concentrations of IL-5 could not reverse the reduced survival rates. After treatment with anti-CD30 Ab, gel electrophoresis of DNA extracted from the eosinophils demonstrated changes consistent with apoptosis. The immobilized F(ab')(2) of the anti-CD30 Ab failed to induce eosinophil apoptosis. The addition of anti-CD18 Ab also completely abrogated the induction of eosinophil apoptosis. Further examination using specific signal transduction inhibitors suggested the involvement of p38, mitogen activated protein kinase kinase 1/2, and specific tyrosine kinase, but not NF kappaB, in the induction of CD30-mediated eosinophil apoptosis. These data demonstrate that CD30 can modify eosinophil survival by causing an extremely rapid and intense induction of apoptosis through a tightly regulated intracellular signaling pathway. PMID- 14764686 TI - Resistance of short term activated T cells to CD95-mediated apoptosis correlates with de novo protein synthesis of c-FLIPshort. AB - In the early phase of an immune response, T cells are activated and acquire effector functions. Whereas these short term activated T cells are resistant to CD95-mediated apoptosis, activated T cells in prolonged culture are readily sensitive, leading to activation-induced cell death and termination of the immune response. The translation inhibitor, cycloheximide, partially overcomes the apoptosis resistance of short term activated primary human T cells. Using this model we show in this study that sensitization of T cells to apoptosis occurs upstream of mitochondria. Neither death-inducing signaling complex formation nor expression of Bcl-2 proteins is altered in sensitized T cells. Although the caspase-8 inhibitor c-FLIP(long) was only slightly down-regulated in sensitized T cells, c-FLIP(short) became almost undetectable. This correlated with caspase-8 activation and apoptosis. These data suggest that c-FLIP(short), rather than c FLIP(long), confers resistance of T cells to CD95-mediated apoptosis in the context of immune responses. PMID- 14764687 TI - IL-10-conditioned dendritic cells, decommissioned for recruitment of adaptive immunity, elicit innate inflammatory gene products in response to danger signals. AB - Dendritic cells (DCs) are the professional APCs of the immune system, enabling T cells to perceive and respond appropriately to potentially dangerous microbes, while also being able to maintain T cell tolerance toward self. In part, such tolerance can be determined by IL-10 released from certain types of regulatory T cells. IL-10 has previously been shown to render DCs unable to activate T cells and it has been assumed that this process represents a general block in maturation. Using serial analysis of gene expression, we show that IL-10 pretreatment of murine bone marrow-derived DCs alone causes significant changes in gene expression. Furthermore, these cells retain the ability to respond to Toll-like receptor agonists, but in a manner skewed toward the selective induction of mediators known to enhance local inflammation and innate immunity, among which we highlight a novel CXCR2 ligand, DC inflammatory protein-1. These data suggest that, while the presence of a protolerogenic and purportedly anti inflammatory agent such as IL-10 precludes DCs from acquiring their potential as initiators of adaptive immunity, their ability to act as initiators of innate immunity in response to Toll-like receptor signaling is enhanced. PMID- 14764688 TI - The cytoplasmic domain of Ig alpha is necessary and sufficient to support efficient early B cell development. AB - The B cell receptor complex (BcR) is essential for normal B lymphocyte function, and surface BcR expression is a crucial checkpoint in B cell development. However, functional requirements for chains of the BcR during development remain controversial. We have used retroviral gene transfer to introduce components of the BcR into chicken B cell precursors during embryonic development. A chimeric heterodimer, in which the cytoplasmic domains of chicken Igalpha and Igbeta are expressed by fusion with the extracellular and transmembrane domains of murine CD8alpha and CD8beta, respectively, targeted the cytoplasmic domains of the BcR to the cell surface in the absence of extracellular BcR domains. Expression of this chimeric heterodimer supported all early stages of embryo B cell development: bursal colonization, clonal expansion, and induction of repertoire diversification by gene conversion. Expression of the cytoplasmic domain of Igalpha, in the absence of the cytoplasmic domain of Igbeta, was not only necessary, but sufficient to support B cell development as efficiently as the endogenous BcR. In contrast, expression of the cytoplasmic domain of Igbeta in the absence of the cytoplasmic domain of Igalpha failed to support B cell development. The ability of the cytoplasmic domain of Igalpha to support early B cell development required a functional Igalpha immunoreceptor tyrosine-based activation motif. These results support a model in which expression of surface IgM following productive V(D)J recombination in developing B cell precursors serves to chaperone the cytoplasmic domain of Igalpha to the B cell surface, thereby initiating subsequent stages of development. PMID- 14764689 TI - HIV-infected Langerhans cells preferentially transmit virus to proliferating autologous CD4+ memory T cells located within Langerhans cell-T cell clusters. AB - Langerhans cells (LC) are likely initial targets for HIV following sexual exposure to virus and provide an efficient means for HIV to gain access to lymph node T cells. The purpose of this study was to examine the nature of the CD4(+) T cell that becomes infected by HIV-infected LC. We infected human LC within tissue explants ex vivo and then, 3 days later, cocultured HIV-infected LC with different subsets of autologous CD4(+) T cells. Using multicolor flow cytometric analyses of LC-CD4(+) T cell cocultures, we documented that HIV-infected LC preferentially infected memory (as compared with naive) CD4(+) T cells. Proliferating and HIV-infected CD4(+) memory T cells were more frequently detected in conjugates of LC and autologous CD4(+) T cells, suggesting that T cells become activated and preferentially get infected through cluster formation with infected LC, rather than getting infected with free virus produced by single HIV-infected LC or T cells. p24(+) Memory CD4(+) T cells proliferated well in the absence of superantigen; by contrast, p24(+) T cells did not divide or divided only once in the presence of staphylococcal enterotoxin B, suggesting that virus production was rapid and induced apoptosis in these cells before significant proliferation could occur. These results highlight that close interactions between dendritic cells, in this case epidermal LC, and T cells are important for optimal HIV replication within specific subsets of CD4(+) T cells. Disrupting cluster formation between LC and memory CD4(+) T cells may be a novel strategy to interfere with sexual transmission of HIV. PMID- 14764690 TI - WSX-1 and glycoprotein 130 constitute a signal-transducing receptor for IL-27. AB - The recently discovered cytokine IL-27 belongs to the IL-6/IL-12 family of cytokines and induced proliferation of naive CD4(+) T cells and the generation of a Th1-type adaptive immune response. Although binding of IL-27 to the cytokine receptor WSX-1 was demonstrated, this interaction proved insufficient to mediate cellular effects. Hence, IL-27 was believed to form a heteromeric signaling receptor complex with WSX-1 and another, yet to be identified, cytokine receptor subunit. In this study, we describe that WSX-1 together with gp130 constitutes a functional signal-transducing receptor for IL-27. We show that neither of the two subunits itself is sufficient to mediate IL-27-induced signal transduction, but that the combination of both is required for this event. Expression analysis of WSX-1 and gp130 by quantitative PCR suggests that IL-27 might have a variety of cellular targets besides naive CD4(+) T cells: we demonstrate gene induction of a subset of inflammatory cytokines in primary human mast cells and monocytes in response to IL-27 stimulation. Thus, IL-27 not only contributes to the development of an adaptive immune response through its action on CD4(+) T cells, it also directly acts on cells of the innate immune system. PMID- 14764691 TI - Identification of new antigenic peptide presented by HLA-Cw7 and encoded by several MAGE genes using dendritic cells transduced with lentiviruses. AB - Antigens encoded by MAGE genes are of particular interest for cancer immunotherapy because they are tumor specific and shared by tumors of different histological types. Several clinical trials are in progress with MAGE peptides, proteins, recombinant poxviruses, and dendritic cells (DC) pulsed with peptides or proteins. The use of gene-modified DC would offer the major advantage of a long-lasting expression of the transgene and a large array of antigenic peptides that fit into the different HLA molecules of the patient. In this study, we tested the ability of gene-modified DC to prime rare Ag-specific T cells, and we identified a new antigenic peptide of clinical interest. CD8(+) T lymphocytes from an individual without cancer were stimulated with monocyte-derived DC, which were infected with a second-generation lentiviral vector encoding MAGE-3. A CTL clone was isolated that recognized peptide EGDCAPEEK presented by HLA-Cw7 molecules, which are expressed by >40% of Caucasians. Interestingly, this new tumor-specific antigenic peptide corresponds to position 212-220 of MAGE-2, -3, 6, and -12. HLA-Cw7 tumor cell lines expressing one of these MAGE genes were lysed by the CTL, indicating that the peptide is efficiently processed in tumor cells and can therefore be used as target for antitumoral vaccination. The risk of tumor escape due to appearance of Ag-loss variants should be reduced by the fact that the peptide is encoded by several MAGE genes. PMID- 14764692 TI - Antigen nonspecific suppression of T cell responses by activated stimulation refractory CD4+ T cells. AB - Several classes of anergic T cells are capable of suppressing naive T cell proliferation and thereby limiting immune responses. Activated T cells, although not anergic, are transiently refractory to restimulation with Ag. We examine in this study whether activated refractory murine T cells can also suppress naive T cell responses. We find that they can, and that they exhibit many of the suppressive properties of anergic T cells. The activated cells strongly diminish Ag-mediated T cell proliferation, an activity that correlates with their refractory period. Suppression is independent of APC numbers and requires cell contact or proximity. Naive T cells stimulated in the presence of activated refractory cells up-regulate CD25 and CD69, but fail to produce IL-2. The addition of IL-2 to culture medium, however, does not prevent the suppression, which is therefore not solely due to the absence of this growth factor. Persistence of the suppressor cells is also not essential. T cells stimulated in their presence and then isolated from them and cultured do not divide. The suppressive cells, however, do not confer a refractory or anergic state on the target T lymphocytes, which can fully respond to antigenic stimulation if removed from the suppressors. Our results therefore provide evidence that activated T cells act as transient suppressor cells, severely constraining bystander T cell stimulation and thereby restricting their response. These results have potentially broad implications for the development and regulation of immune responses. PMID- 14764693 TI - The impact of thymic antigen diversity on the size of the selected T cell repertoire. AB - The TCR repertoire of a normal animal is shaped in the thymus by ligand-specific positive- and negative-selection events. These processes are believed to be determined at the single-cell level primarily by the affinity of the TCR-ligand interactions. The relationships among all the variables involved are still unknown due to the complexity of the interactions and the lack of quantitative analysis of those parameters. In this study, we developed a quantitative model of thymic selection that provides estimates of the fractions of positively and negatively selected thymocytes in the cortex and in the medulla, as well as upper bound ranges for the number of selecting ligands required for the generation of a normal diverse TCR repertoire. Fitting the model to current estimates of positive and negative-selected thymocytes leads to specific predictions. The results indicate the following: 1) the bulk of thymocyte death takes place in the cortex, and it is due to neglect; 2) the probability of a thymocyte to be negatively selected in the cortex is at least 10-fold lower than in the medulla; 3) <60 ligands are involved in cortical positive selection; and 4) negative selection in the medulla is constrained by a large diversity of selecting ligands on medullary APCs. PMID- 14764694 TI - Human cytomegalovirus proteins pp65 and immediate early protein 1 are common targets for CD8+ T cell responses in children with congenital or postnatal human cytomegalovirus infection. AB - Recombinant modified vaccinia Ankara- and peptide-based IFN-gamma ELISPOT assays were used to detect and measure human CMV (HCMV)-specific CD8(+) T cell responses to the pp65 (UL83) and immediate early protein 1 (IE1; UL123) gene products in 16 HCMV-infected infants and children. Age at study ranged from birth to 2 years. HCMV-specific CD8(+) T cells were detected in 14 (88%) of 16 children at frequencies ranging from 60 to >2000 spots/million PBMC. Responses were detected as early as 1 day of age in infants with documented congenital infection. Nine children responded to both pp65 and IE1, whereas responses to pp65 or IE1 alone were detected in three and two children, respectively. Regardless of the specificity of initial responses, IE1-specific responses predominated by 1 year of age. Changes in HCMV epitopes targeted by the CD8(+) T cell responses were observed over time; epitopes commonly recognized by HLA-A2(+) adults with latent HCMV infection did not fully account for responses detected in early childhood. Finally, the detection of HCMV-specific CD8(+) T cell responses was temporally associated with a decrease in peripheral blood HCMV load. Taken altogether, these data demonstrate that the fetus and young infant can generate virus-specific CD8(+) T cell responses. Changes observed in the protein and epitope-specificity of HCMV-specific CD8(+) T cells over time are consistent with those observed after other primary viral infections. The temporal association between the detection of HCMV-specific CD8(+) T cell responses and the reduction in blood HCMV load supports the importance of CD8(+) T cells in controlling primary HCMV viremia. PMID- 14764695 TI - NF-kappa B controls cell fate specification, survival, and molecular differentiation of immunoregulatory natural T lymphocytes. AB - Ontogenetic, homeostatic, and functional deficiencies within immunoregulatory natural T (iNKT) lymphocytes underlie various inflammatory immune disorders including autoimmunity. Signaling events that control cell fate specification and molecular differentiation of iNKT cells are only partly understood. Here we demonstrate that these processes within iNKT cells require classical NF-kappaB signaling. Inhibition of NF-kappaB signaling blocks iNKT cell ontogeny at an immature stage and reveals an apparent, novel precursor in which negative selection occurs. Most importantly, this block occurs due to a lack of survival signals, as Bcl-x(L) overexpression rescues iNKT cell ontogeny. Maturation of immature iNKT cell precursors induces Bcl-2 expression, which is defective in the absence of NF-kappaB signaling. Bcl-x(L) overexpression also rescues this maturation-induced Bcl-2 expression. Thus, antiapoptotic signals relayed by NF kappaB critically control cell fate specification and molecular differentiation of iNKT cells and, hence, reveal a novel role for such signals within the immune system. PMID- 14764696 TI - Adjuvant costimulation during secondary antigen challenge directs qualitative aspects of oral tolerance induction, particularly during the neonatal period. AB - In this report we demonstrate that although passive feeding of specific Ag to mice as neonates or adults can induce oral tolerance in both the cellular and humoral arms of the immune response, quantitative and, in particular, qualitative aspects of the tolerance process are determined by the nature of the inflammatory costimuli provided at the time of secondary Ag challenge. Moreover, this dependency upon nonspecific costimulation is more profound in Ag-fed neonates than in their adult counterparts. Thus, administration of Ag in the Th1-selective adjuvant CFA to prefed animals resulted in significant inhibition of IgG2a, IL-2, and IFN-gamma responses, whereas IL-5 responses were increased. In contrast, rechallenge with Ag in the Th2-selective adjuvant aluminum hydroxide resulted in significant inhibition of IgG1, IgE, IL-2, and IL-5 responses, whereas IFN-gamma responses were increased. Additionally, although soluble Ag challenge of prefed adults revealed marginal tolerogenic effects, the same challenge protocol in animals prefed as neonates elicited enhanced Th2-dependent IgG1 production. These results suggest that inflammatory stimulation at the time of Ag challenge is obligatory to trigger oral tolerance mechanisms, particularly in animals fed as neonates and also that the type of adjuvant used at the time of challenge selects for the type of Th cell population to be inhibited. PMID- 14764697 TI - CD4+ T cells regulate CD8+ T cell-mediated cutaneous immune responses by restricting effector T cell development through a Fas ligand-dependent mechanism. AB - The magnitude and duration of CD8(+) T cell-mediated responses in the skin to hapten sensitization and challenge, contact hypersensitivity (CHS), is negatively regulated by CD4(+) T cells through an unknown mechanism. In this study we show that CD4(+) T cells restrict the development and expansion of hapten-specific CD8(+) T cells mediating CHS responses to 2,4-dinitrofluorobenzene. In the absence of CD4(+) T cells, high numbers of hapten-specific CD8(+) T cells producing IFN-gamma were detected in the skin-draining lymph nodes on day 5 postsensitization, and these numbers decreased slightly, but were maintained through day 9, correlating with the increased magnitude and duration of CHS responses observed in these mice. In the presence of CD4(+) T cells, the number of hapten-specific CD8(+) T cells producing IFN-gamma detected on day 5 postsensitization was lower and quickly fell to background levels by day 7. The limited development of effector CD8(+) T cells was associated with decreased numbers of hapten-presenting dendritic cells in the lymphoid priming site. This form of immunoregulation was absent after sensitization of Fas ligand-defective gld mice. Transfer of wild-type CD4(+) T cells to gld mice restored the negative regulation of CD8(+) T cell priming and the immune response to hapten challenge in gld-recipient mice. These results indicate that CD4(+) T cells restrict hapten specific CD8(+) T cell priming for CHS responses through a Fas ligand-dependent mechanism. PMID- 14764698 TI - Reduced thymocyte development in sonic hedgehog knockout embryos. AB - The Hedgehog family of secreted intercellular signaling molecules are regulators of patterning and organogenesis during animal development. In this study we provide genetic evidence that Sonic Hedgehog (Shh) has a role in the control of murine T cell development. Analysis of Shh(-/-) mouse embryos revealed that Shh regulates fetal thymus cellularity and thymocyte differentiation. Shh is necessary for expansion of CD4(-)CD8(-) double-negative (DN) thymocytes and for efficient transition from the earliest CD44(+)CD25(-) DN population to the subsequent CD44(+)CD25(+) DN population and from DN to CD4(+)CD8(+) double positive cells. PMID- 14764699 TI - Dendritic cell maturation requires STAT1 and is under feedback regulation by suppressors of cytokine signaling. AB - In this study we show that activation of STAT pathways is developmentally regulated and plays a role in dendritic cell (DC) differentiation and maturation. The STAT6 signaling pathway is constitutively activated in immature DC (iDC) and declines as iDCs differentiate into mature DCs (mDCs). However, down-regulation of this pathway during DC differentiation is accompanied by dramatic induction of suppressors of cytokine signaling 1 (SOCS1), SOCS2, SOCS3, and cytokine-induced Src homology 2-containing protein expression, suggesting that inhibition of STAT6 signaling may be required for DC maturation. In contrast, STAT1 signaling is most robust in mDCs and is not inhibited by the up-regulated SOCS proteins, indicating that STAT1 and STAT6 pathways are distinctly regulated in maturing DC. Furthermore, optimal activation of STAT1 during DC maturation requires both IL-4 and GM-CSF, suggesting that synergistic effects of both cytokines may in part provide the requisite STAT1 signaling intensity for DC maturation. Analyses of STAT1(-/-) DCs reveal a role for STAT1 in repressing CD86 expression in precursor DCs and up-regulating CD40, CD11c, and SOCS1 expression in mDCs. We further show that SOCS proteins are differentially induced by IL-4 and GM-CSF in DCs. SOCS1 is primarily induced by IL-4 through a STAT1-dependent mechanism, whereas SOCS3 is induced mainly by GM-CSF. Taken together, these results suggest that cytokine induced maturation of DCs is under feedback regulation by SOCS proteins and that the switch from constitutive activation of the STAT6 pathway in iDCs to predominant use of STAT1 signals in mDC is mediated in part by STAT1-induced SOCS expression. PMID- 14764700 TI - Caspase-3 is a component of Fas death-inducing signaling complex in lipid rafts and its activity is required for complete caspase-8 activation during Fas mediated cell death. AB - Since its discovery, caspase-8 has been placed at the apex of the proteolytic cascade triggered by death receptor (DR) cross-linking. Because of its capacity to interact with the cytoplasmic portion of DR, it has been suggested that caspase-8 acts independently of other caspases in the initiation of Fas and other DR signaling. In this study, we demonstrate that in Jurkat cells, caspase-3 cleavage is an early step during Fas-induced apoptosis. We show that caspase-3 processing into its p20 occurs rapidly after Fas cross-linking, in the absence of mitochondrial depolarization and caspase-9 activation. Moreover, caspase-3 is present in lipid rafts of untreated Jurkat cells and peripheral T lymphocytes. Caspase-3, caspase-8, and Fas-associated death domain are further recruited to lipid rafts of Jurkat cells following anti-Fas treatment. Fas immunoprecipitation reveals that caspase-3 is a component of the death-inducing signaling complex, suggesting that this cysteine protease is in close proximity to caspase-8. Furthermore, transduction of Jurkat cells with a caspase-3 dominant-negative form inhibits caspase-8 processing and results in inhibition of apoptosis, suggesting that caspase-3 activity is required for caspase-8 activation. Overall, these findings support a model whereby caspase-3 is a component of the death-inducing signaling complex located in lipid rafts, and as such, is involved in the amplification of caspase-8 activity by the mitochondrion. PMID- 14764701 TI - The impact of self-tolerance on the polyclonal CD8+ T cell repertoire. AB - TCRs possess considerable cross-reactivity toward structurally related Ags. Because the signaling threshold for negative selection is lower than that required for activation of mature T cells, the question arises as to which extent thymic deletion of self-specific T cells affects T cell responsiveness toward foreign peptides. In this study we show, in three different mouse models systems, that the polyclonal CD8(+) T cell repertoire has a marked ability to react against the majority of Ags related to self despite self-tolerance, even in cases where self and foreign differ only marginally at a single TCR-contact residue. Thus, while individual T cells are markedly cross-reactive, the ability to distinguish between closely related Ags is introduced at the polyclonal T cell level. PMID- 14764702 TI - Activation of discoidin domain receptor 1 isoform b with collagen up-regulates chemokine production in human macrophages: role of p38 mitogen-activated protein kinase and NF-kappa B. AB - Macrophages produce an array of proinflammatory mediators at sites of inflammation and contribute to the development of inflammatory responses. Important roles for cytokines, such as IL-1 or TNF-alpha, and bacterial products, such as LPS, in this process have been well documented; however, the role for the extracellular matrix proteins, such as collagen, remains unclear. We previously reported that discoidin domain receptor 1 (DDR1), a nonintegrin collagen receptor, is expressed during differentiation of human monocytes into macrophages, and the interaction of the DDR1b isoform with collagen facilitates their differentiation via the p38 mitogen-activated protein kinase (MAPK) pathway. In this study, we report that the interaction of DDR1b with collagen up regulates the production of IL-8, macrophage inflammatory protein-1alpha, and monocyte chemoattractant protein-1 in human macrophages in a p38 MAPK- and NF kappaB-dependent manner. p38 MAPK was critical for DDR1b-mediated, increased NF kappaB trans-activity, but not for IkappaB degradation or NF-kappaB nuclear translocation, suggesting a role for p38 MAPK in the modification of NF-kappaB. DDR1b-mediated IkappaB degradation was mediated through the recruitment of the adaptor protein Shc to the LXNPXY motif of the receptor and the downstream TNFR associated factor 6/NF-kappaB activator 1 signaling cascade. Taken together, our study has identified NF-kappaB as a novel target of DDR1b signaling and provided a novel mechanism by which tissue-infiltrating macrophages produce large amounts of chemokines during the development of inflammatory diseases. Intervention of DDR1b signaling may be useful to control inflammatory diseases in which these proteins play an important role. PMID- 14764703 TI - Immunosuppressive activity of endovanilloids: N-arachidonoyl-dopamine inhibits activation of the NF-kappa B, NFAT, and activator protein 1 signaling pathways. AB - Endogenous N-acyl dopamines such as N-arachidonoyldopamine (NADA) and N oleoyldopamine have been recently identified as a new class of brain neurotransmitters sharing endocannabinoid and endovanilloid biological activities. As endocannabinoids show immunomodulatory activity, and T cells play a key role in the onset of several diseases that affect the CNS, we have evaluated the immunosuppressive activity of NADA and N-oleoyldopamine in human T cells, discovering that both compounds are potent inhibitors of early and late events in TCR-mediated T cell activation. Moreover, we found that NADA specifically inhibited both IL-2 and TNF-alpha gene transcription in stimulated Jurkat T cells. To further characterize the inhibitory mechanisms of NADA at the transcriptional level, we examined the DNA binding and transcriptional activities of NF-kappaB, NF-AT, and AP-1 transcription factors in Jurkat cells. We found that NADA inhibited NF-kappaB-dependent transcriptional activity without affecting either degradation of the cytoplasmic NF-kappaB inhibitory protein, IkappaBalpha, or DNA binding activity. However, phosphorylation of the p65/RelA subunit was clearly inhibited by NADA in stimulated cells. In addition, NADA inhibited both binding to DNA and the transcriptional activity of NF-AT and AP-1, as expected from the inhibition of NF-AT1 dephosphorylation and c-Jun N-terminal kinase activation in stimulated T cells. Finally, overexpression of a constitutively active form of calcineurin demonstrated that this phosphatase may represent one of the main targets of NADA. These findings provide new mechanistic insights into the anti-inflammatory activities of NADA and highlight their potential to design novel therapeutic strategies to manage inflammatory diseases. PMID- 14764704 TI - Molecular characterization of human 4Ig-B7-H3, a member of the B7 family with four Ig-like domains. AB - In an effort to characterize molecules with immunoregulatory potential, we raised mAbs to human dendritic cells. We selected an Ab that recognizes a molecule that is induced on monocytes differentiated in vitro toward dendritic cells. Retroviral expression cloning identified this molecule as B7-H3, a member of the B7 family described recently. In contrast to an earlier report, in which B7-H3 was described as a molecule consisting of two Ig-like domains, our cDNA encoded a type I membrane protein with four Ig-like domains, and the molecule identified by us was therefore named 4Ig-B7-H3. mRNA analysis as well as Western blotting experiments performed by us did not reveal evidence for a small B7-H3. B7-H3 is not expressed on peripheral blood lymphocytes, monocytes, or granulocytes. Upon in vitro stimulation, the expression of B7-H3 is induced on T cells, B cells, and NK cells. A number of different approaches were used to investigate the function of human B7-H3. In contrast to an earlier report, our data do not support a costimulatory role of B7-H3 in anti-CD3-mediated activation of the TCR-complex resulting in T cell proliferation and IFN-gamma production. PMID- 14764706 TI - Conformation and glycosylation of a megalin fragment correlate with nephritogenicity in Heymann nephritis. AB - Active Heymann nephritis (AHN), a rat model of autoimmune glomerulonephritis, is induced by immunization with autologous megalin, a 600-kDa cell surface glycoprotein isolated from crude renal extracts. Recombinant proteins containing a 563-residue N-terminal sequence of megalin were obtained from Escherichia coli and baculovirus-insect cell expression systems. Rats immunized with the soluble, secreted protein encoded by a baculovirus construct elicited high titer anti megalin autoantibodies and developed glomerular immune deposits and elevated proteinuria consistent with AHN. Rats treated with the bacterial or nonsecreted insect cell proteins produced a milder anti-megalin response and did not develop the disease. Nephritogenicity appeared to correlate with conformational or other structural features of native megalin. All three recombinant proteins were reactive in Western blots with rabbit anti-megalin antiserum, whereas the insect cell-derived proteins reacted preferentially in Western blot and ELISA with anti megalin autoantibodies from rats with AHN induced by native megalin. Only the secreted insect cell product was stained in a lectin blot, suggesting its specific glycosylation. These observations provide evidence that a megalin N terminal domain includes B and T cell epitopes sufficient for a pathogenic autoimmune response and that a native-like conformation and glycosylation are essential for the induction of disease. The importance of conformational B cell epitopes for pathogenic autoantibodies recapitulates observations made in other models of organ-specific autoimmune disease. Glycosidic modifications could influence the presentation of either B or T cell epitopes in AHN, consistent with emerging evidence of the role of post-translational modifications in pathogenic autoimmune responses. PMID- 14764705 TI - Biases in Ig lambda light chain rearrangements in human intestinal plasma cells. AB - Human intestinal lamina propria plasma cells are considered to be the progeny of chronically stimulated germinal centers located in organized gut-associated lymphoid tissues such as Peyer's patches and isolated lymphoid follicles. We have sampled human colonic lamina propria plasma cells and naive and memory B cell subsets from human Peyer's patches by microdissection of immunohistochemically stained tissue sections and used PCR methods and sequence analysis to compare IgVlambdaJlambda rearrangements in the plasma cell and B cell populations. Rearrangements that were either in-frame or out-of-frame between V and J were compared. Usage of IgVlambda families in the in-frame rearrangements from the plasma cells resembled that observed in the mantle cells, suggesting that antigenic selection for cellular specificity does not dramatically favor any particular Vlambda segment. However, in marked contrast, out-of-frame rearrangements involving Vlambda1 and Vlambda2 families are rarely observed in intestinal plasma cells, whereas rearrangements involving Vlambda5 are increased. This resulted in significantly biased ratios of in-frame:out-of-frame rearrangements in these Vlambda families. Out-of-frame rearrangements of IgVlambdaJlambda from plasma cells, including those involving the Vlambda5 family, have a significant tendency not to involve Jlambda1, consistent with the hypothesis that this population includes rearrangements generated by secondary recombination events. We propose that modification of out-of-frame rearrangements of IgVlambdaJlambda exists, probably a consequence of secondary rearrangements. This may be a mechanism to avoid translocations to susceptible out-of-frame IgVlambdaJlambda rearrangements during somatic hypermutation. PMID- 14764707 TI - Fc epsilon RI gamma-ITAM is differentially required for mast cell function in vivo. AB - The cross-linking of IgE-bound FcepsilonRI by Ags triggers mast cell activation leading to allergic reactions. The in vivo contribution of FcepsilonRIgamma signaling to IgE/FcepsilonRI-mediated mast cell responses has not yet been elucidated. In this study FcepsilonRIgamma(-/-) mast cells were reconstituted with either wild-type or mutant FcepsilonRIgamma in transgenic mice and transfected mast cells in vitro. We demonstrate that FcepsilonRIgamma immunoreceptor tyrosine-based activation motif is essential for degranulation, cytokine production, and PG synthesis as well as for passive systemic anaphylaxis. Recent reports have suggested that cell surface FcepsilonRI expression and mast cell survival are regulated by IgE in the absence of Ag, although the molecular mechanism is largely unknown. We also found that the promotion of mast cell survival by IgE without Ags is mediated by signals through the FcepsilonRIgamma-immunoreceptor tyrosine-based activation motif. In contrast, the IgE-mediated up-regulation of FcepsilonRI is independent of FcepsilonRIgamma signaling. These results indicate that FcepsilonRIgamma-mediated signals differentially regulate the receptor expression, activation, and survival of mast cells and systemic anaphylaxis. PMID- 14764708 TI - The crystal structure of human CD1b with a bound bacterial glycolipid. AB - The human MHC class I-like molecule CD1b is distinctive among CD1 alleles in that it is capable of presenting a set of glycolipid species that show a very broad range of variation in the lengths of their acyl chains. A structure of CD1b complexed with relatively short acyl chain glycolipids plus detergent suggested how an interlinked network of channels within the Ag-binding groove could accommodate acyl chain lengths of up to 80 carbons. The structure of CD1b complexed with glucose monomycolate, reported in this study, confirms this hypothesis and illustrates how the distinctive substituents of intracellular bacterial glycolipids can be accommodated. The Ag-binding groove of CD1b is, uniquely among CD1 alleles, partitioned into channels suitable for the compact accommodation of lengthy acyl chains. The current crystal structure illustrates for the first time the binding of a natural bacterial lipid Ag to CD1b and shows how its novel structural features fit this molecule for its role in the immune response to intracellular bacteria. PMID- 14764709 TI - Activation/division of lymphocytes results in increased levels of cytoplasmic activation/proliferation-associated protein-1: prototype of a new family of proteins. AB - We purified from activated T lymphocytes a novel, highly conserved, 116-kDa, intracellular protein that occurred at high levels in the large, dividing cells of the thymus, was up-regulated when resting T or B lymphocytes or hemopoietic progenitors were activated, and was down-regulated when a monocytic leukemia, M1, was induced to differentiate. Expression of the protein was highest in the thymus and spleen and lowest in tissues with a low proportion of dividing cells such as kidney or muscle, although expression was high in the brain. The protein was localized to the cytosol and was phosphorylated, which is consistent with a previous report that the Xenopus laevis ortholog was phosphorylated by a mitotically activated kinase (1 ). The cDNA was previously mischaracterized as encoding p137, a 137-kDa GPI-linked membrane protein (2 ). We propose that the authentic protein encoded by this cDNA be called cytoplasmic activation/proliferation-associated protein-1 (caprin-1), and show that it is the prototype of a novel family of proteins characterized by two novel protein domains, termed homology regions-1 and -2 (HR-1, HR-2). Although we have found evidence for caprins only in urochordates and vertebrates, two insect proteins exhibit well-conserved HR-1 domains. The HR-1 and HR-2 domains have no known function, although the HR-1 of caprin-1 appeared necessary for formation of multimeric complexes of caprin-1. Overexpression of a fusion protein of enhanced green fluorescent protein and caprin-1 induced a specific, dose-dependent suppression of the proliferation of NIH-3T3 cells, consistent with the notion that caprin-1 plays a role in cellular activation or proliferation. PMID- 14764710 TI - Antibody-dependent cellular cytotoxicity via humoral immune epitope of Nef protein expressed on cell surface. AB - Antibodies against various proteins of HIV type 1 (HIV-1) can be detected in HIV 1-infected individuals. We previously reported that the level of Ab response against one Nef epitope is correlated with HIV-1 disease progression. To elucidate the mechanism for this correlation, we examined Ab-dependent cellular cytotoxicity (ADCC) against target cells expressing Nef. We observed efficient cytotoxicity against Nef-expressing target cells in the presence of patient plasma and PBMCs. This ADCC activity was correlated with the dilution of plasma from HIV-1-infected patients. Addition of a specific synthetic peptide (peptide 31:FLKEKGGLE) corresponding to the Nef epitope reduced cell lysis to approximately 50%. These results suggest that PBMCs of HIV-1-infected patients may exert ADCC via anti-Nef Abs in the patients' own plasma and serve as a mechanism used by the immune system to regulate HIV-1 replication. PMID- 14764711 TI - Macrophages, CD4+ or CD8+ cells are each sufficient for protection against Chlamydia pneumoniae infection through their ability to secrete IFN-gamma. AB - By using a T, B, or NK cell-deficient mouse strain (recombinase-activating gene (RAG)-1(-/-)/common cytokine receptor gamma-chain (gamma(C)R)), and T and B cell and IFN-gamma-deficient (RAG-1(-/-)/IFN-gamma(-/-)) mice, we have studied the generation of immunity against infection by Chlamydia pneumoniae. We found that IFN-gamma secreted by innate-cell populations protect against C. pneumoniae infection. However, NK cells were not needed for such IFN-gamma-dependent innate immune protection. Inoculation of wild type, but not IFN-gamma(-/-) bone marrow derived macrophages protected RAG-1(-/-)/IFN-gamma(-/-) mice against C. pneumoniae infection. In line, pulmonary macrophages from RAG-1(-/-) C. pneumoniae-infected mice expressed IFN-gamma mRNA. Reconstitution of RAG-1(-/ )/gamma(c)R(-/-) or RAG-1(-/-)/IFN-gamma(-/-) mice with CD4(+) or CD8(+) cells by i.v. transfer of FACS sorted wild type spleen cells (SC) increased resistance to C. pneumoniae infection. On the contrary, no protection was observed upon transfer of IFN-gamma(-/-) CD4(+) or IFN-gamma(-/-) CD8(+) SC. T cell-dependent protection against C. pneumoniae was weaker when IFN-gammaR(-/-) CD4(+) or IFN gammaR(-/-) CD8(+) SC were inoculated into RAG-1(-/-)/IFN-gamma(-/-) mice. Thus both nonlymphoid and T cell-derived IFN-gamma can play a central and complementary role in protection against C. pneumoniae. IFN-gamma secreted by nonlymphoid cells was not required for T cell-mediated protection against C. pneumoniae; however, IFN-gamma regulated T cell protective functions. PMID- 14764712 TI - Calcium and its role in the nuclear translocation and activation of cytosolic phospholipase A2 in cells rendered sensitive to TNF-induced apoptosis by cycloheximide. AB - In these experiments, we investigated the role of calcium as a second messenger in the apoptotic activation of cytosolic phospholipase A(2) (cPLA(2)). As our model, we used a murine fibroblast cell line (C3HA) that was induced to undergo apoptosis by a combination of TNF and cycloheximide. Using fura 2 Ca(2+) imaging, we found strong evidence for an intracellular calcium response after 1 h of treatment, which correlated with the onset of phosphatidylserine externalization, but preceded effector procaspase processing by several hours. The response was strongest in the perinuclear region, where mean levels rose 83% (144 +/- 14 nM in untreated cells vs 264 +/- 39 nM in treated), while cells displaying morphological evidence of apoptosis had the highest levels of calcium (250-1000 nM). Verapamil blocked this response, indicating an extracellular source for the calcium. Fluorescence microscopy revealed a pattern of nuclear translocation of cPLA(2) during apoptosis, which was also blocked by verapamil, indicating an important role for calcium in this process. In addition, we found that verapamil prevented the release of [(3)H]arachidonic acid from C3HA cells induced to undergo apoptosis by the chemotherapeutic agents vinblastine, melphalan, and cis platinum. Together, these data suggest that calcium is important for cPLA(2) activation by diverse apoptotic stimuli. PMID- 14764713 TI - Peripheral CD4 T cells rapidly accumulate at the host: parasite interface during an inflammatory Th2 memory response. AB - Memory peripheral Th2 immune responses to infectious pathogens are not well studied due to the lack of suitable models and the difficulty of assessing Th2 cytokine expression at sites of inflammation. We have examined the localized immune response to a nematode parasite that encysts in the small intestine. An unexpected architecture was observed on day 4 of the memory response, with granulocytes and macrophages infiltrating the cyst and CD4(+), TCR-alphabeta(+) T cells surrounding the cyst. Laser capture microdissection analysis showed a pronounced CD4-dependent Th2 cytokine pattern at the cyst region only during the memory response, demonstrating that the Th2 memory response is readily distinguished from the primary response by the rapid accumulation of Th2 effector cells at the host:parasite interface. PMID- 14764714 TI - Haemophilus influenzae type b-outer membrane protein complex glycoconjugate vaccine induces cytokine production by engaging human toll-like receptor 2 (TLR2) and requires the presence of TLR2 for optimal immunogenicity. AB - Conjugate vaccines consisting of the capsular polysaccharide (PS) of Haemophilus influenzae type b (Hib) covalently linked to carrier proteins, unlike pure PS, are immunogenic in infants and have significantly reduced Hib infections in the United States, but require multiple doses to induce protective anti-PS Ab titers. Hib-meningococcal outer membrane protein complex (OMPC) conjugate vaccine, however, elicits protective anti-PS Ab titers after one dose. We found that OMPC and Hib-OMPC engaged human Toll-like receptor 2 (TLR2) expressed in human embryonic kidney (HEK) cells, inducing IL-8 production, and engaged mouse TLR2 on bone marrow-derived dendritic cells, inducing TNF release. Hib conjugated to the carrier proteins CRM(197) and tetanus toxoid did not engage TLR2 on HEK or dendritic cells. Engagement of TLR2 by Hib-OMPC was MyD88 dependent, as Hib-OMPC induced TNF production was ablated in MyD88 knockout (KO) mice. Hib-OMPC was significantly less immunogenic in TLR2 KO mice, inducing lower Hib PS IgG and IgM titers compared with those in wild-type mice. Splenocytes from OMPC-immunized TLR2 KO mice also produced significantly less IL-6 and TNF-alpha than those from wild-type mice. Hib-OMPC is unique among glycoconjugate vaccines by engaging TLR2, and the ability of Hib-OMPC to elicit protective levels of Abs after one dose may be related to TLR2-mediated induction and regulation of cytokines produced by T cells and macrophages in addition to the peptide/MHC II-dependent recruitment of T cell help commonly afforded by carrier proteins. TLR2 engagement by an adjuvant or carrier protein may be a useful strategy for augmentation of the anti-PS Ab response induced by glycoconjugate vaccines. PMID- 14764715 TI - Parasite antigen-driven basophils are a major source of IL-4 in human filarial infections. AB - Basophil contribution to the IL-4 pool in filarial infections was assessed using PBMC from 20 patients with active filarial infections and from 9 uninfected subjects. Patient basophils released histamine in response to Brugia malayi Ag (BmAg). They also released IL-4 within 2 h after exposure to BmAg, as assessed by intracellular cytokine flow cytometry. This IL-4 induction was Ag specific, as IL 4 was not detected in BmAg-exposed basophils obtained from uninfected subjects. Although there were, on average, 64 times more CD4(+) T cells than basophils in the peripheral circulation of filaria-infected patients, the absolute numbers of basophils and CD4(+) T cells producing IL-4 per 100000 PBMC were equivalent (geometric mean: 16 IL-4-producing basophils/100000 PBMC vs 22 IL-4-producing CD4(+) T cells/100000 PBMC). Basophils also released IL-4 in response to both low and high concentrations of BmAg, whereas CD4(+) T cells released IL-4 only after incubation with a high concentration of BmAg, raising the possibility that basophils, due to their lower threshold for activation, may actually release IL-4 more frequently than CD4(+) T cells in vivo. Furthermore, IL-4 production in vitro by Ag-stimulated purified basophils or CD4(+) T cells provided evidence that basophils release greater quantities of IL-4 per cell than CD4(+) T cells in response to BmAg. These results suggest that, when Ag-specific IgE is present in a filaria-infected individual, basophils function to amplify the ongoing Th2 response by releasing IL-4 in greater amounts and possibly more frequently than CD4(+) T cells in response to filarial Ag. PMID- 14764716 TI - The zinc finger mutation C417R of I-kappa B kinase gamma impairs lipopolysaccharide- and TNF-mediated NF-kappa B activation through inhibiting phosphorylation of the I-kappa B kinase beta activation loop. AB - The activation of the I-kappaB kinase (IKK) complex by TNF or LPS stimulates phosphorylation and degradation of I-kappaBalpha, leading to the nuclear translocation of NF-kappaB. The IKK complex is mainly composed of two catalytic subunits, IKKalpha and IKKbeta, and a chaperon subunit IKKgamma. Although IKKgamma does not have catalytic activity, it is essential for IKK activation induced by multiple stimuli. Importantly, the key residue cysteine 417 at the zinc finger domain of IKKgamma has been found to be mutated to arginine (IKKgammaC417R) in a human genetic disorder called the anhydrotic ectodermal dysplasia with immunodeficiency. To understand the underlying mechanisms of immunodeficiency, we examined whether the IKKgammaC417R mutant modified IKK activation and NF-kappaB transcription stimulated by LPS or TNF in human monocytes. We found that overexpression of IKKgammaC417R severely impaired LPS- and TNF-induced I-kappaBalpha phosphorylation and degradation in a dominant negative fashion. Also, LPS- and TNF-induced NF-kappaB transcription was inhibited by IKKgammaC417R. The reconstitution of IKKgamma, but not IKKgammaC417R, in IKKgamma-deficient cells restored NF-kappaB signaling, indicating the zinc finger structure of IKKgamma plays a key role in IKK activation. Moreover, C417R mutation in IKKgamma abolished both LPS- and TNF induced phosphorylation of the activation loop of IKKbeta. Collectively, our results indicated that the zinc finger structure of IKKgamma plays a key role in LPS- and TNF-induced NF-kappaB activation. The anhydrotic ectodermal dysplasia with immunodeficiency patients' immunodeficiency may be associated with NF-kappaB defect in response to bacterial stimulation. PMID- 14764717 TI - Recognition of homo- and heterosubtypic variants of influenza A viruses by human CD8+ T lymphocytes. AB - In the present study, the recognition of epitope variants of influenza A viruses by human CTL was investigated. To this end, human CD8(+) CTL clones, specific for natural variants of the HLA-B*3501-restricted epitope in the nucleoprotein (NP(418-426)), were generated. As determined in (51)Cr release assays and by flow cytometry with HLA-B*3501-peptide tetrameric complexes, CTL clones were found to be specific for epitopes within one subtype or cross-reactive with heterosubtypic variants of the epitope. Using eight natural variants of the epitope, positions in the 9-mer important for T cell recognition and involved in escape from CTL immunity were identified and visualized using multidimensional scaling. It was shown that positions 4 and 5 in the 9-mer epitope were important determinants of T cell specificity. The in vivo existence of CD8(+) cells cross-reactive with homo- and heterosubtypic variants of the epitope was further confirmed using polyclonal T cell populations obtained after stimulation of PBMC with different influenza A viruses. Based on the observed recognition patterns of the clonal and polyclonal T cell populations and serology, it is hypothesized that consecutive infections with influenza viruses containing different variants of the epitope select for cross-reactive T cells in vivo. PMID- 14764718 TI - Protective immunization against group B meningococci using anti-idiotypic mimics of the capsular polysaccharide. AB - Use of the serogroup B meningococcal capsular polysaccharide (MenB CP) as a vaccine is hampered by the presence of epitopes that cross-react with human polysialic acid. As non-cross-reactive, protective capsular epitopes have also been described, we set out to develop protein mimics of one of such epitopes using as a template a highly protective mAb (mAb Seam 3) raised against a chemically modified form of the MenB CP (N-Pr MenB CP). Using phage display, anti idiotypic single-chain Ab fragments (scFvs) were obtained from spleen cells of mice immunized with the Seam 3 mAb. Two Seam 3-specific scFvs competed with N-Pr MenB CP for binding to either mAb Seam 3 or rabbit Abs present in typing sera. Moreover, in mice and rabbits the scFvs elicited the production of Abs reacting with both N-Pr MenB CP and whole meningococci, but not with human polysialic acid. These scFv-induced Ab responses were boostable and of the Th1 type, as shown by a predominance of IgG2a. In addition, passive immunization with sera from scFv-immunized animals partially protected neonatal mice from experimental infection with group B meningococci. In conclusion, we have produced anti idiotypic scFvs that mimic a protective MenB CP epitope and may be useful in the development of an alternative group B meningococcal vaccine. PMID- 14764719 TI - Cyclooxygenase-2-mediated prostaglandin E2 production in mesenteric lymph nodes and in cultured macrophages and dendritic cells after infection with Salmonella. AB - Although numerous studies have demonstrated the ability of intestinal epithelial cells to produce PGs after infection with wild-type strains of Salmonella, few studies have focused on Salmonella-induced prostanoids in mucosal lymphoid tissues. This is surprising in view of the profound effects PGs can have on the host response. To begin to address PG production at mucosal sites, mice were orally inoculated with Salmonella, and at varying times postinfection cyclooxygenase-2 (COX-2) mRNA expression and PGE(2) synthesis were investigated. COX-2 mRNA expression was highly inducible in the mesenteric lymph nodes, whereas COX-1 mRNA levels were constitutive. PGE(2) production also increased significantly in the mesenteric lymph nodes following exposure to viable Salmonella, but not after exposure to killed bacteria. This increased PGE(2) response could be blocked by treatment of mice with the selective COX-2 inhibitor, celecoxib. Treatment of mice with celecoxib during salmonellosis resulted in increased viable bacteria in the mesenteric lymph nodes by day 3 postinfection. However, celecoxib treatment prolonged the survival of lethally infected animals. In vitro studies demonstrated Salmonella-induced up-regulation of COX-2 mRNA expression and PGE(2) secretion by both macrophages and dendritic cells, which could also be blocked in the presence of celecoxib. Interestingly, exposure of these cultured APCs to viable Salmonella was a much greater stimulus for induction of PGE(2) synthesis than exposure to Salmonella-derived LPS. The present study demonstrates induction of PGE(2) synthesis in mesenteric lymph nodes, macrophages, and dendritic cells after infection with wild-type salmonella. PMID- 14764720 TI - HIV-1-specific memory CD4+ T cells are phenotypically less mature than cytomegalovirus-specific memory CD4+ T cells. AB - HIV-1-specific CD4(+) T cells are qualitatively dysfunctional in the majority of HIV-1-infected individuals and are thus unable to effectively control viral replication. The current study extensively details the maturational phenotype of memory CD4(+) T cells directed against HIV-1 and CMV. We find that HIV-1-specific CD4(+) T cells are skewed to an early central memory phenotype, whereas CMV specific CD4(+) T cells generally display a late effector memory phenotype. These differences hold true for both IFN-gamma- and IL-2-producing virus-specific CD4(+) T cells, are present during all disease stages, and persist even after highly active antiretroviral therapy (HAART). In addition, after HAART, HIV-1 specific CD4(+) T cells are enriched for CD27(+)CD28(-)-expressing cells, a rare phenotype, reflecting an early intermediate stage of differentiation. We found no correlation between differentiation phenotype of HIV-1-specific CD4(+) T cells and HIV-1 plasma viral load or HIV-1 disease progression. Surprisingly, HIV-1 viral load affected the maturational phenotype of CMV-specific CD4(+) T cells toward an earlier, less-differentiated state. In summary, our data indicate that the maturational state of HIV-1-specific CD4(+) T cells cannot be a sole explanation for loss of containment of HIV-1. However, HIV-1 replication can affect the phenotype of CD4(+) T cells of other specificities, which might adversely affect their ability to control those pathogens. The role for HIV-1 specific CD4(+) T cells expressing CD27(+)CD28(-) after HAART remains to be determined. PMID- 14764721 TI - Protective T cell immunity against malaria liver stage after vaccination with live sporozoites under chloroquine treatment. AB - In this study we present the first systematic analysis of the immunity induced by normal Plasmodium yoelii sporozoites in mice. Immunization with sporozoites, which was conducted under chloroquine treatment to minimize the influence of blood stage parasites, induced a strong protection against a subsequent sporozoite and, to a lesser extent, against infected RBC challenges. The protection induced by this immunization protocol proved to be very effective. Induction of this protective immunity depended on the presence of liver stage parasites, as primaquine treatment concurrent with sporozoite immunization abrogated protection. Protection was not found to be mediated by the Abs elicited against pre-erythrocytic and blood stage parasites, as demonstrated by inhibition assays of sporozoite penetration or development in vitro and in vivo assays of sporozoite infectivity or blood stage parasite development. CD4(+) and CD8(+) T cells were, however, responsible for the protection through the induction of IFN gamma and NO. PMID- 14764722 TI - Liver dendritic cells present bacterial antigens and produce cytokines upon Salmonella encounter. AB - The capacity of murine liver dendritic cells (DC) to present bacterial Ags and produce cytokines after encounter with Salmonella was studied. Freshly isolated, nonparenchymal liver CD11c(+) cells had heterogeneous expression of MHC class II and CD11b and a low level of CD40 and CD86 expression. Characterization of liver DC subsets revealed that CD8alpha(-)CD4(-) double negative cells constituted the majority of liver CD11c(+) ( approximately 85%) with few cells expressing CD8alpha or CD4. Flow cytometry analysis of freshly isolated CD11c(+) cells enriched from the liver and cocultured with Salmonella expressing green fluorescent protein (GFP) showed that CD11c(+) MHC class II(high) cells had a greater capacity to internalize Salmonella relative to CD11c(+) MHC class II(low) cells. Moreover, both CD8alpha(-) and CD8alpha(+) liver DC internalized bacteria with similar efficiency after both in vitro and in vivo infection. CD11c(+) cells enriched from the liver could also process Salmonella for peptide presentation on MHC class I and class II to primary, Ag-specific T cells after internalization requiring actin cytoskeletal rearrangements. Flow cytometry analysis of liver CD11c(+) cells infected with Salmonella expressing GFP showed that both CD8alpha( ) and CD8alpha(+) DC produced IL-12p40 and TNF-alpha. The majority of cytokine positive cells did not contain bacteria (GFP(-)) whereas only a minor fraction of cytokine-positive cells were GFP(+). Furthermore, only approximately 30-50% of liver DC containing bacteria (GFP(+)) produced cytokines. Thus, liver DC can internalize and process Salmonella for peptide presentation to CD4(+) and CD8(+) T cells and elicit proinflammatory cytokine production upon Salmonella encounter, suggesting that DC in the liver may contribute to immunity against hepatotropic bacteria. PMID- 14764723 TI - Suppression of osteoprotegerin expression by prostaglandin E2 is crucially involved in lipopolysaccharide-induced osteoclast formation. AB - LPS is a potent stimulator of bone resorption in inflammatory diseases. The mechanism by which LPS induces osteoclastogenesis was studied in cocultures of mouse osteoblasts and bone marrow cells. LPS stimulated osteoclast formation and PGE(2) production in cocultures of mouse osteoblasts and bone marrow cells, and the stimulation was completely inhibited by NS398, a cyclooxygenase-2 inhibitor. Osteoblasts, but not bone marrow cells, produced PGE(2) in response to LPS. LPS induced osteoclast formation was also inhibited by osteoprotegerin (OPG), a decoy receptor of receptor activator of NF-kappaB ligand (RANKL), but not by anti-mouse TNFR1 Ab or IL-1 receptor antagonist. LPS induced both stimulation of RANKL mRNA expression and inhibition of OPG mRNA expression in osteoblasts. NS398 blocked LPS-induced down-regulation of OPG mRNA expression, but not LPS-induced up regulation of RANKL mRNA expression, suggesting that down-regulation of OPG expression by PGE(2) is involved in LPS-induced osteoclast formation in the cocultures. NS398 failed to inhibit LPS-induced osteoclastogenesis in cocultures containing OPG knockout mouse-derived osteoblasts. IL-1 also stimulated PGE(2) production in osteoblasts and osteoclast formation in the cocultures, and the stimulation was inhibited by NS398. As seen with LPS, NS398 failed to inhibit IL 1-induced osteoclast formation in cocultures with OPG-deficient osteoblasts. These results suggest that IL-1 as well as LPS stimulates osteoclastogenesis through two parallel events: direct enhancement of RANKL expression and suppression of OPG expression, which is mediated by PGE(2) production. PMID- 14764724 TI - TNF receptor signaling contributes to chemokine secretion, inflammation, and respiratory deficits during Pneumocystis pneumonia. AB - CD8(+) T cells contribute to the pathophysiology of Pneumocystis pneumonia (PcP) in a murine model of AIDS-related disease. The present studies were undertaken to more precisely define the mechanisms by which these immune cells mediate the inflammatory response that leads to lung injury. Experimental mice were depleted of either CD4(+) T cells or both CD4(+) and CD8(+) T cells and then infected with Pneumocystis: The CD4(+)-depleted mice had significantly greater pulmonary TNF alpha levels than mice depleted of both CD4(+) and CD8(+) T cells. Elevated TNF alpha levels were associated with increased lung concentrations of the chemokines RANTES, monocyte chemoattractant protein 1, macrophage-inflammatory protein 2, and cytokine-induced neutrophil chemoattractant. To determine whether TNFR signaling was involved in the CD8(+) T cell-dependent chemokine response, TNFRI- and II-deficient mice were CD4(+) depleted and infected with Pneumocystis: TNFR deficient mice had significantly reduced pulmonary RANTES, monocyte chemoattractant protein 1, macrophage-inflammatory protein 2, and cytokine induced neutrophil chemoattractant responses, reduced inflammatory cell recruitment to the alveoli, and reduced histological evidence of PcP-related alveolitis as compared with infected wild-type mice. Diminished pulmonary inflammation correlated with improved surfactant activity and improved pulmonary function in the TNFR-deficient mice. These data indicate that TNFR signaling is required for maximal CD8(+) T cell-dependent pulmonary inflammation and lung injury during PcP and also demonstrate that CD8(+) T cells can use TNFR signaling pathways to respond to an extracellular fungal pathogen. PMID- 14764725 TI - Involvement of reactive oxygen species in Toll-like receptor 4-dependent activation of NF-kappa B. AB - Although oxidative stress has been thought to play a general role in the activation of NF-kappaB, the involvement of reactive oxygen species (ROS) in facilitating nuclear translocation of NF-kappaB in neutrophils has not been described. In addition, the mechanisms by which ROS modulate the transcriptional activity of NF-kappaB in response to Toll-like receptor 4 (TLR4)-dependent signaling are not well characterized. To examine these issues, oxidant-dependent signaling events downstream of TLR4 were investigated in neutrophils stimulated with LPS. Pretreatment of neutrophils with the antioxidants N-acetylcysteine or alpha-tocopherol prevented LPS-induced nuclear translocation of NF-kappaB. Antioxidant treatment of LPS-stimulated neutrophils also inhibited the production of proinflammatory cytokines (TNF-alpha, macrophage inflammatory protein-2, and IL-1beta), as well as activation of the kinases IkappaB kinase alpha, IkappaB kinase beta, p38, Akt, and extracellular receptor-activated kinases 1 and 2. The decrease in cytoplasmic levels of IkappaBalpha produced by exposure of neutrophils to LPS was prevented by N-acetylcysteine or alpha-tocopherol. Activation of IL-1R-associated kinase-1 (IRAK-1) and IRAK-4 in response to LPS stimulation was inhibited by antioxidants. These results demonstrate that proximal events in TLR4 signaling, at or antecedent to IRAK-1 and IRAK-4 activation, are oxidant dependent and indicate that ROS can modulate NF-kappaB dependent transcription through their involvement in early TLR4-mediated cellular responses. PMID- 14764726 TI - B7-DC regulates asthmatic response by an IFN-gamma-dependent mechanism. AB - B7-H1 (PD-L1) and B7-DC (PD-L2) are the ligands for programmed death-1 (PD-1), which is a member of the CD28/CTLA-4 family and has been implicated in peripheral tolerance. We investigated the roles of B7-H1 and B7-DC in a murine OVA-induced allergic asthma model. B7-H1 was constitutively expressed on dendritic cells, macrophages, B cells, and T cells in the lungs of naive mice, and its expression could be dramatically increased after allergen challenge. In contrast, B7-DC expression was scarcely expressed on dendritic cells in naive mice, but was up regulated after allergen challenge, although the up-regulation of B7-DC expression on macrophages was minimal. Treatment of mice with anti-B7-DC mAb at the time of allergen challenge, but not at the time of sensitization, significantly increased their airway hyper-reactivity and eosinophilia. Such treatment also resulted in the increased production of IL-5 and IL-13, and decreased IFN-gamma production in the lungs and draining lymph node cells. These changes were diminished when mice were depleted of IFN-gamma by anti-IFN-gamma mAb pretreatment. Interestingly, treatment with anti-B7-H1 or anti-PD-1 mAb did not significantly affect the asthmatic response. These results suggest a unique role for B7-DC in the regulation of asthmatic response through an IFN-gamma dependent, but PD-1-independent, mechanism. PMID- 14764727 TI - Role of bradykinin B2 and B1 receptors in the local, remote, and systemic inflammatory responses that follow intestinal ischemia and reperfusion injury. AB - The administration of bradykinin may attenuate ischemia and reperfusion (I/R) injury by acting on B(2)Rs. Blockade of B(2)R has also been shown to ameliorate lesions associated with I/R injury. In an attempt to explain these contradictory results, the objective of the present work was to investigate the role of and interaction between B(1) and B(2) receptors in a model of intestinal I/R injury in mice. The bradykinin B(2)R antagonist (HOE 140) inhibited reperfusion-induced inflammatory tissue injury and delayed lethality. After I/R, there was an increase in the expression of B(1)R mRNA that was prevented by HOE 140. In mice that were deficient in B(1)Rs (B(1)R(-/-) mice), inflammatory tissue injury was abrogated, and lethality was delayed and partially prevented. Pretreatment with HOE 140 reversed the protective anti-inflammatory and antilethality effects provided by the B(1)R(-/-) phenotype. Thus, B(2)Rs are a major driving force for B(1)R activation and consequent induction of inflammatory injury and lethality. In contrast, activation of B(2)Rs may prevent exacerbated tissue injury and lethality, an effect unmasked in B(1)R(-/-) mice and likely dependent on the vasodilatory actions of B(2)Rs. Blockade of B(1)Rs could be a more effective strategy than B(2) or B(1)/B(2) receptor blockade for the treatment of the inflammatory injuries that follow I/R. PMID- 14764728 TI - Contribution of antigen-primed CD8+ T cells to the development of airway hyperresponsiveness and inflammation is associated with IL-13. AB - The role of Th2/CD4 T cells, which secrete IL-4, IL-5, and IL-13, in allergic disease is well established; however, the role of CD8(+) T cells (allergen induced airway hyperresponsiveness (AHR) and inflammation) is less clear. This study was conducted to define the role of Ag-primed CD8(+) T cells in the development of these allergen-induced responses. CD8-deficient (CD8(-/-)) mice and wild-type mice were sensitized to OVA by i.p. injection and then challenged with OVA via the airways. Compared with wild-type mice, CD8(-/-) mice developed significantly lower airway responsiveness to inhaled methacholine and lung eosinophilia, and exhibited decreased IL-13 production both in vivo, in the bronchoalveolar lavage (BAL) fluid, and in vitro, following Ag stimulation of peribronchial lymph node (PBLN) cells in culture. Reconstitution of sensitized and challenged CD8(-/-) mice with allergen-sensitized CD8(+) T cells fully restored the development of AHR, BAL eosinophilia, and IL-13 levels in BAL and in culture supernatants from PBLN cells. In contrast, transfer of naive CD8(+) T cells or allergen-sensitized CD8(+) T cells from IL-13-deficient donor mice failed to do so. Intracellular cytokine staining of lung as well as PBLN T cells revealed that CD8(+) T cells were a source of IL-13. These data suggest that Ag primed CD8(+) T cells are required for the full development of AHR and airway inflammation, which appears to be associated with IL-13 production from these primed T cells. PMID- 14764729 TI - Microtubule-associated serine/threonine kinase-205 kDa and Fc gamma receptor control IL-12 p40 synthesis and NF-kappa B activation. AB - Stimulation of murine macrophages with LPS results in the coordinated activation of a set of proinflammatory cytokines and costimulatory molecules, including TNF alpha, IL-6, IL-1, IL-8, IL-12, and CD80. Macrophage LPS-induced synthesis of IL 12 is inhibited following FcgammaR ligation; TNF-alpha secretion is unchanged. We report that microtubule-associated serine/threonine kinase-205 kDa (MAST205) is required for LPS-induced IL-12 synthesis. RNA interference-mediated suppression of MAST205 results in the inhibition of LPS-stimulated IL-12 promoter activity and IL-12 secretion, from both J774 cells and bone marrow-derived macrophages. Similarly, dominant-negative MAST205 mutants inhibit LPS-stimulated IL-12 synthesis and NF-kappaB activation, but do not affect IL-1 or TNF-alpha signaling. Finally, macrophage FcgammaR ligation regulates MAST205 by inducing the rapid ubiquitination and proteasomal degradation of the protein. PMID- 14764730 TI - Inhibition of c-Jun NH2-terminal kinase activity improves ischemia/reperfusion injury in rat lungs. AB - Although c-Jun NH(2)-terminal kinase (JNK) has been implicated in the pathogenesis of transplantation-induced ischemia/reperfusion (I/R) injury in various organs, its significance in lung transplantation has not been conclusively elucidated. We therefore attempted to measure the transitional changes in JNK and AP-1 activities in I/R-injured lungs. Subsequently, we assessed the effects of JNK inhibition by the three agents including SP600125 on the degree of lung injury assessed by means of various biological markers in bronchoalveolar lavage fluid and histological examination including detection of apoptosis. In addition, we evaluated the changes in p38, extracellular signal regulated kinase, and NF-kappaB-DNA binding activity. I/R injury was established in the isolated rat lung preserved in modified Euro-Collins solution at 4 degrees C for 4 h followed by reperfusion at 37 degrees C for 3 h. We found that AP-1 was transiently activated during ischemia but showed sustained activation during reperfusion, leading to significant lung injury and apoptosis. The change in AP-1 was generally in parallel with that of JNK, which was activated in epithelial cells (bronchial and alveolar), alveolar macrophages, and smooth muscle cells (bronchial and vascular) on immunohistochemical examination. The change in NF kappaB qualitatively differed from that of AP-1. Protein leakage, release of lactate dehydrogenase and TNF-alpha into bronchoalveolar lavage fluid, and lung injury were improved, and apoptosis was suppressed by JNK inhibition. In conclusion, JNK plays a pivotal role in mediating lung injury caused by I/R. Therefore, inhibition of JNK activity has potential as an effective therapeutic strategy for preventing I/R injury during lung transplantation. PMID- 14764731 TI - Peptide-mediated inhibition of neutrophil transmigration by blocking CD47 interactions with signal regulatory protein alpha. AB - CD47, a cell surface transmembrane Ig superfamily member, is an extracellular ligand for signal regulatory protein (SIRPalpha). Interactions between CD47 and SIRPalpha regulate many important immune cell functions including neutrophil (PMN) transmigration. Here we report identification of a novel function-blocking peptide, CERVIGTGWVRC, that structurally mimics an epitope on CD47 and binds to SIRPalpha. The CERVIGTGWVRC sequence was identified by panning phage display libraries on the inhibitory CD47 mAb, C5D5. In vitro PMN migration assays demonstrated that peptide CERVIGTGWVRC specifically inhibited PMN migration across intestinal epithelial monolayers and matrix in a dose-dependent fashion. Further studies using recombinant proteins indicated that the peptide specifically blocks CD47 and SIRPalpha binding in a dose-dependent fashion. Protein binding assays using SIRPalpha domain-specific recombinant proteins demonstrated that this peptide directly bound to the distal-most Ig loop of SIRPalpha, the same loop where CD47 binds. In summary, these findings support the relevance of CD47-SIRPalpha interactions in regulation of PMN transmigration and provide structural data predicting the key residues involved on the surface of CD47. Such peptide reagents may be useful for studies on experimental models of inflammation and provide a template for the design of anti-inflammatory agents. PMID- 14764732 TI - Matrix metalloproteinase-9 deficiency results in enhanced allergen-induced airway inflammation. AB - Matrix metalloproteinases (MMPs) are a large family of endopeptidases that proteolytically degrade extracellular matrix. Many different cells produce MMP-9, and levels have been shown to be up-regulated in patients with allergic asthma. The aim of this study was to investigate the in vivo role of MMP-9 during allergen-induced airway inflammation. Acute allergic pulmonary eosinophilia was established in MMP-9 knockout (KO) and wild-type (WT) control mice by sensitization and challenge with OVA. Cell recruitment was significantly increased in both bronchoalveolar lavage (BAL) and lung tissue compartments in MMP-9 KO mice compared with WT mice. This heightened cell recruitment was primarily due to increased eosinophils and Th2 cells in the BAL and lung tissue of MMP-9 KO mice in comparison with WT controls. Moreover, levels of the Th2 cytokines, IL-4 and IL-13, and the chemokines eotaxin/CCL11 and macrophage derived chemokine/CCL22 were substantially increased in MMP-9 KO mice compared with WT after OVA challenge. Resolution of eosinophilia was similar between MMP-9 KO and WT mice, but Th2 cells persisted in BAL and lungs of MMP-9 KO mice for longer than in WT mice. Our results indicate that MMP-9 is critically involved in the recruitment of eosinophils and Th2 cells to the lung following allergen challenge, and suggest that MMP-9 plays a role in the development of Th2 responses to allergen. PMID- 14764733 TI - Inflammatory gene profiles in gastric mucosa during Helicobacter pylori infection in humans. AB - Helicobacter pylori infection is associated with an inflammatory response in the gastric mucosa, ultimately leading to cellular hyperproliferation and malignant transformation. Hitherto, only expression of a single gene, or a limited number of genes, has been investigated in infected patients. cDNA arrays were therefore used to establish the global pattern of gene expression in gastric tissue of healthy subjects and of H. pylori-infected patients. Two main gene expression profiles were identified based on cluster analysis. The data obtained suggest a strong involvement of selected Toll-like receptors, adhesion molecules, chemokines, and ILs in the mucosal response. This pattern is clearly different from that observed using gastric epithelial cell lines infected in vitro with H. pylori. The presence of a "Helicobacter-infection signature," i.e., a set of genes that are up-regulated in biopsies from H. pylori-infected patients, could be derived from this analysis. The genotype of the bacteria (presence of genes encoding cytotoxin-associated Ag, vacuolating cytotoxin, and blood group Ag binding adhesin) was analyzed by PCR and shown to be associated with differential expression of a subset of genes, but not the general gene expression pattern. The expression data of the array hybridization was confirmed by quantitative real time PCR assays. Future studies may help identify gene expression patterns predictive of complications of the infection. PMID- 14764734 TI - Key roles of CD4+ T cells and IFN-gamma in the development of abdominal aortic aneurysms in a murine model. AB - Abdominal aortic aneurysm (AAA) is one of a number of diseases associated with a prominent inflammatory cell infiltrate and local destruction of structural matrix macromolecules. This inflammatory infiltrate is predominately composed of T lymphocytes and macrophages. Delineating specific contribution of these inflammatory cells and their cytokines in AAA formation is the key to understanding AAA and other chronic inflammatory disease processes. Our previous studies have demonstrated that macrophages are the major source of matrix metalloproteinase-9, which is required for aneurysmal degeneration in the murine AAA model. However, the role of CD4(+) T cells, the most abundant infiltrates in aneurysmal aortic tissue, is uncertain. In the present study, we found that in the absence of CD4(+) T cells, mice are resistant to aneurysm induction. Previous studies have shown that IFN-gamma levels are increased in AAA. IFN-gamma is a main product of T cells. Intraperitoneal IFN-gamma was able to partially reconstitute aneurysms in CD4(-/-) mice. Furthermore, mice with a targeted deletion of IFN-gamma have attenuation of MMP expression and inhibition of aneurysm development. Aneurysms in IFN-gamma(-/-) mice can be reconstituted by reinfusion of competent splenocytes from the corresponding wild-type mice. This study demonstrates the pivotal role that T cells and the T cell cytokine, IFN gamma, play in orchestrating matrix remodeling in AAA. This study has important implications for other degenerative diseases associated with matrix destruction. PMID- 14764735 TI - Inhibition of IL-10 receptor function in alveolar macrophages by Toll-like receptor agonists. AB - Despite an immunosuppressive lung environment, alveolar macrophages (AM) retain the capacity to respond to microorganisms. This report demonstrates that IL-10, constitutively produced by normal alveolar epithelium, stimulates signal transduction through the IL-10R on AM and that IL-10R function can be inhibited by stimulation of Toll-like receptor (TLR) on AM. IL-10 mRNA and protein were constitutively expressed in normal alveolar epithelium of mice, and IL-10R were constitutively expressed on normal murine AM. Stimulation of AM through TLR2, TLR4, or TLR9 was sufficient to inhibit IL-10R signal transduction, including phosphorylation and nuclear translocation of STAT3 transcription factor. Inhibition of IL-10R function by TLRs was not associated with a decrease in IL 10R expression, but did require expression of the myeloid differentiation factor 88 adaptor protein. Continuous exposure of macrophages to IL-10 caused sustained expression of the chemokine receptors CCR1 and CCR5. However, the addition of TLR ligands inhibited IL-10-induced expression of CCR1 and CCR5. Finally, exposure of macrophages to TLR ligands blocked the ability of IL-10 to inhibit the induction of TNF-alpha by C2-ceramide. These findings demonstrate a novel regulatory mechanism that may allow AM to overcome inhibitory effects of constitutive IL-10 in the lungs that may permit a more effective response to pulmonary infections. PMID- 14764736 TI - Construction of hevein (Hev b 6.02) with reduced allergenicity for immunotherapy of latex allergy by comutation of six amino acid residues on the conformational IgE epitopes. AB - Recently we have established that IgE Abs bind to conformational epitopes in the N- and C-terminal regions of the major natural rubber latex allergen, hevein (Hev b 6.02). To identify the critical amino acid residues that interact with IgE, the hevein sequence was scanned by using site-specific mutations. Twenty-nine hevein mutants were designed and produced by a baculovirus expression system in insect cells and tested by IgE inhibition-ELISA using sera from 26 latex allergic patients. Six potential IgE-interacting residues of hevein (Arg(5), Lys(10), Glu(29), Tyr(30), His(35), and Gln(38)) were identified and characterized further in detail. Based on these six residues, two triple mutants (Hdelta3A, Hdelta3B) and hevein mutant where all six residues were mutated (Hdelta6), were designed, modeled, and produced. Structural and functional properties of these combinatory mutants were compared experimentally and in silico with those of recombinant hevein. The IgE-binding affinity of the mutants decreased by three to five orders of magnitude as compared with that of recombinant hevein. Skin prick test reactivity of the triple mutant HDelta3A was drastically reduced and that of the six-residue mutant Hdelta6 was completely abolished in all patients examined in this study. The approach presented in this paper offers tools for identification and modification of amino acid residues on conformational epitopes of allergens that interact with IgE. Hevein with a highly reduced ability to bind IgE should provide a valuable candidate molecule for immunotherapy of latex allergy and is anticipated to have a low risk of systemic side effects. PMID- 14764737 TI - Role of Toll-like receptor 4 in endotoxin-induced acute renal failure. AB - Toll-like receptor 4 (TLR4) is present on monocytes and other cell types, and mediates inflammatory events such as the release of TNF after exposure to LPS. C3H/HeJ mice are resistant to LPS-induced mortality, due to a naturally occurring mutation in TLR4. We therefore hypothesized that LPS-induced acute renal failure (ARF) requires systemic TNF release triggered by LPS acting on extrarenal TLR4. We injected C3H/HeJ mice and C3H/HeOuJ controls with 0.25 mg of LPS, and sacrificed them 6 h later for analysis of blood urea nitrogen (BUN) and kidney tissue (n = 8 per group). In contrast to C3H/HeOuJ controls, C3H/HeJ mice were completely resistant to LPS-induced ARF (6-h BUN of 32.3 +/- 1.1 vs 61.7 +/- 5.6 mg/dl). C3H/HeJ mice released no TNF into the circulation at 2 h (0.00 vs 1.24 +/ 0.16 ng/ml), had less renal neutrophil infiltration (6.4 +/- 1.0 vs 11.4 +/- 1.3 neutrophils per high power field), and less renal apoptosis, as assessed by DNA laddering. Transplant studies showed that C3H/HeJ recipients of wild-type kidneys (n = 9) were protected from LPS-induced ARF, while wild-type recipients of C3H/HeJ kidneys (n = 11) developed severe LPS-induced ARF (24-h BUN 44.0 +/- 4.1 vs 112.1 +/- 20.0 mg/dl). These experiments support our hypothesis that LPS acts on extrarenal TLR4, thereby leading to systemic TNF release and subsequent ARF. Renal neutrophil infiltration and renal cell apoptosis are potential mechanisms by which endotoxemia leads to functional ARF. PMID- 14764738 TI - Respective roles of decay-accelerating factor and CD59 in circumventing glomerular injury in acute nephrotoxic serum nephritis. AB - Decay-accelerating factor (DAF or CD55) and CD59 are regulators that protect self cells from C3b deposition and C5b-9 assembly on their surfaces. Their relative roles in protecting glomeruli in immune-mediated renal diseases in vivo are unknown. We induced nephrotoxic serum (NTS) nephritis in Daf1(-/-), CD59a(-/-), Daf1(-/-)CD59a(-/-), and wild-type (WT) mice by administering NTS IgG. After 18 h, we assessed proteinuria, and performed histological, immunohistochemical, and electron microscopic analyses of kidneys. Twenty-four mice in each group were studied. Baseline albuminuria in the Daf1(-/-), CD59a(-/-), and Daf1(-/-)CD59a(-/ ) mice was 82, 83, and 139 as compared with 92 microg/mg creatinine in the WT controls (p > 0.1). After NTS, albuminuria in CD59a(-/-) and WT mice (186 +/- 154 and 183 +/- 137 microg/mg creatinine, p > 0.1) was similar. In contrast, Daf1(-/ ) mice developed severe albuminuria (378 +/- 520, p < 0.05) that was further exacerbated in Daf1(-/-)CD59a(-/-) mice (577 +/- 785 micro g/mg creatinine, p < 0.05). Glomerular histology showed essentially no infiltrating leukocytes in any group. In contrast, electron microscopy revealed prominent podocyte foot process effacement in Daf1(-/-) mice with more widespread and severe damage in the double knockouts compared with only mild focal changes in CD59a(-/-) or WT mice. In all animals, deposition of administered (sheep) NTS Ig was equivalent. This contrasted with marked deposition of both C3 and C9 in Daf1(-/-)CD59a(-/-) and Daf1(-/-) mice, which was evident as early as 2 h post-NTS injection. The results support the proposition that in autoantibody-mediated nephritis, DAF serves as the primary barrier to classical pathway-mediated injury, while CD59 limits consequent C5b-9-mediated cell damage. PMID- 14764739 TI - TNF-alpha coupled to membrane of apoptotic cells favors the cross-priming to melanoma antigens. AB - The cross-presentation of Ags derived from apoptotic cell processing contributes to peripheral tolerance. Environmental signals possibly modify this default outcome, favoring cross-priming. In this study, we anchored via a biotin-avidin biotin bridge soluble TNF-alpha to the membrane of apoptotic melanoma cells and studied in vivo and in vitro the interaction with Ag-presenting phagocytes. TNF alpha-coated apoptotic melanoma cells injected s.c. had a faster and more efficient access to draining lymph nodes, with cross-priming of melanoma-specific CTLs and delayed outgrowth of melanomas in all treated animals. Twenty percent of the animals, in the absence of further adjuvant, did not develop the tumor. Immature dendritic cells challenged with TNF-alpha-coated apoptotic melanoma cells secreted proinflammatory cytokines in an autocrine/paracrine fashion, efficiently matured, as assessed functionally and by flow cytometry and cross presented with enhanced efficiency melanoma Ags to MHC class I- and II-restricted T cells. The results indicate that TNF-alpha targeted to apoptotic membranes, at concentrations that can be safely reached in growing tumors without undue systemic toxicity, influences the outcome of the disposal of dying cells and enhances tumor immunogenicity. PMID- 14764740 TI - A spontaneous model for autoimmune myocarditis using the human MHC molecule HLA DQ8. AB - Genome-wide analyses have shown that the MHC class II region is the principal locus that confers susceptibility to a number of human autoimmune diseases. Due to the high degree of linkage disequilibrium across the MHC, it has been difficult to dissect the contribution of individual genes to disease susceptibility. As a result, intensive efforts have been made to generate mice transgenic for human class II molecules as models of autoimmune disease. However, in every case, additional manipulations-such as immunization with Ag in adjuvant, expression of immunostimulants on target tissues, or coexpression of TCR transgenes-have been required to induce disease. In this study, we show that expression of the human HLA-DQ8 (DQA1*0301/DQB1*0302) molecule alone in three lines of transgenic nonobese diabetic murine class II-deficient (mII(-/-)) mice results in the spontaneous development of autoimmune myocarditis. The disease shares key features of human myocarditis and was characterized by lymphocytic infiltrates in the myocardium and cardiac myocyte destruction, circulating IgG autoantibodies against cardiac myosin heavy chain, and premature death due to heart failure. We demonstrate that myocarditis could be transferred into healthy HLA-DQ8(+)RAG-1(-/-)mII(-/-) nonobese diabetic recipients with lymphocytes, but not sera. It has been widely thought that autoimmune myocarditis is of infectious etiology, with the immune responses arising secondary to cardiac damage from pathogens. These studies provide direct experimental evidence that spontaneous autoimmune myocarditis can occur in the absence of infection and that expression of HLA-DQ8 confers susceptibility to this organ-specific autoimmune disease. PMID- 14764741 TI - In vivo evidence that peptide vaccination can induce HLA-DR-restricted CD4+ T cells reactive to a class I tumor peptide. AB - Vaccination with class I tumor peptides has been performed to induce tumor reactive CD8(+) T cells in vivo. However, the kinds of immune responses that vaccination might elicit in patients are not fully understood. In this study we tried to elucidate the mechanisms by which vaccination of class I binding tumor peptides into an HLA-A2(+) lung cancer patient elicited dramatic amounts of IgG1 and IgG2 specific to a nonamer peptide, ubiquitin-conjugated enzyme variant Kua (UBE2V)(43-51). The UBE2V(43-51) peptide contains cysteine at the sixth position. HLA-DR-restricted and UBE2V(43-51) peptide-recognizing CD4(+) T cells were induced from postvaccination, but not from prevaccination, PBMCs of the cancer patient. In addition, a CD4(+) T cell line (UB-2) and its clone (UB-2.3), both of which recognize the UBE2V(43-51) peptide in the context of HLA-DRB1*0403 molecules, were successfully established from postvaccination PBMCs. The peptide vaccination increased the frequency of peptide-specific T cells, especially CD4(+) T cells. In contrast, mass spectrometric analysis revealed that the vaccinated UBE2V(43-51) peptide contained both monomeric and dimeric forms. Both forms, fractionated by reverse phase HPLC, were recognized by UB-2 and UB-2.3 cells. Recognition by these CD4(+) T cells was observed despite the addition of a reduction reagent or the fixation of APC. Overall, these results indicate that vaccination with class I tumor peptides can induce HLA-DR-restricted CD4(+) T cells in vivo and elicit humoral immune responses, and that a cysteine-containing peptide can be recognized by CD4(+) T cells not only as a monomer, but also as a dimer. PMID- 14764742 TI - Pulmonary edema fluid from patients with early lung injury stimulates fibroblast proliferation through IL-1 beta-induced IL-6 expression. AB - Although the fibroproliferative response to lung injury occurs with a high frequency in patients with clinical acute lung injury, the mechanisms that initiate this response are largely unknown. This study was undertaken first to identify fibroblast mitogenic factors in pulmonary edema fluid, and second to examine the human lung fibroblast's gene expression profile in response to pulmonary edema fluid. The edema fluid obtained from patients with early lung injury has an eightfold higher concentration of IL-1beta and a twofold greater IL 1beta-dependent mitogenic effect than does fluid obtained from control patients with hydrostatic pulmonary edema. Furthermore, fibroblasts responded to acute lung injury patient-derived edema fluid through production of soluble mediators that possess an autocrine mitogenic effect. Gene array analysis reveals that acute lung injury edema fluid induces several inflammation-modulating and proliferation-related genes in fibroblasts, whose inductions are similarly dependent on bioactive IL-1beta. Most notably, the 20-fold induction of IL-6 mRNA and protein was completely blocked by IL-1 receptor antagonist. The combined addition of IL-1beta and IL-6 was mitogenic, and the proliferative response to conditioned medium from IL-1beta-exposed cells was blocked by antagonistically acting Abs to IL-6 or to gp130. These novel findings indicate that soluble IL 1beta bioactivity and autocrine IL-1beta-dependent IL-6 up-regulation are critical initiators of fibroblast activation and proliferation and that they likely play a role in the fibroproliferative response seen in human acute lung injury. PMID- 14764743 TI - CD40 ligand dysregulation in HIV infection: HIV glycoprotein 120 inhibits signaling cascades upstream of CD40 ligand transcription. AB - IL-12 production and up-regulation of CD40 ligand (CD40L) expression are impaired in the PBMC of HIV-infected donors, and exogenous CD40L rescues IL-12 production by such cells. In this study, we implicate dysregulation of CD40L expression in the IL-12 defect associated with HIV by demonstrating that induction of CD40L expression by anti-CD3/CD28 stimulation was directly correlated with the IL-12 productive capacity of PBMC. Further, we demonstrate marked decreases in the induction of CD40L protein and mRNA following anti-CD3/CD28 stimulation in HIV infected donors compared with uninfected donors, with a tight association between these two levels. Inhibition of CD40L up-regulation was selective, as induction of CD69 or OX40 was not as severely affected. Increased instability of CD40L mRNA did not constitute a major mechanism in CD40L dysregulation, thus suggesting a potential defect in the signaling cascades upstream of transcription. The mechanisms by which HIV infection affects the induction of CD40L expression appear to involve HIV gp120-mediated engagement of CD4. Indeed, anti-CD4 mAb or inactivated HIV virions that harbor a conformationally intact gp120 significantly inhibited CD40L up-regulation at both the protein and mRNA levels. This inhibition was due to the native, virion-associated gp120, as coculture with soluble CD4 or heat treatment of inactivated HIV abolished their effect. These in vitro models mirror the CD40L defect seen in cells from HIV-infected donors and thus provide a suitable model to investigate HIV-induced CD40L dysregulation. Clear elucidation of mechanism(s) may well lead to the development of novel immunotherapeutic approaches to HIV infection. PMID- 14764744 TI - HIV-1 does not provoke alteration of cytokine gene expression in lymphoid tissue after acute infection ex vivo. AB - The cytokine response to invading microorganisms is critical for priming the adaptive immune response. During acute HIV infection, the response is disrupted, but the mechanism is poorly understood. We examined the cytokine response in human lymphoid tissue, acutely infected ex vivo with HIV. Lymphoid tissue was cultured either as blocks or as human lymphocyte aggregate cultures (HLAC) of tonsils and lymph nodes. This approach allowed us to examine the effects of HIV on cytokines using distinct culture techniques. In contrast to HLAC, mock infected tissue blocks displayed a 50- to 100-fold up-regulation of mRNAs for IL 1beta, -6, and -8 in the first 6 days of culture. Parallel increases were also noted at the protein level in the supernatants. Although IL-1beta, -6, and -8 are known to synergistically enhance HIV replication, peak HIV replication (measured as p24 Ag) was similar in tissue blocks and HLAC. Surprisingly, vigorous HIV replication of CXCR4- and CCR5-tropic HIV strains did not result in characteristic mRNA profiles for IL-1beta, -2, -4, -6, -8, -10, -12, -15, IFN gamma, TNF-alpha, TGF-beta, and beta-chemokines in tissue blocks or HLAC. The increased expression of IL-1beta, -6, and -8 in tissue blocks may approximate clinical situations with heightened immune activation; neutralization of these cytokines resulted in inhibition of HIV replication, suggesting that these cytokines may contribute to HIV replication in certain clinical settings. These results also indicate that different molecular mechanisms govern HIV replication in tissue blocks and HLAC. Prevention of effective cytokine responses may be an important mechanism that HIV uses during acute infection. PMID- 14764745 TI - Breakdown of tolerance to a self-peptide of acetylcholine receptor alpha-subunit induces experimental myasthenia gravis in rats. AB - Experimental autoimmune myasthenia gravis (EAMG), a model for human myasthenia (MG), is routinely induced in susceptible rat strains by a single immunization with Torpedo acetylcholine receptor (TAChR). TAChR immunization induces anti-AChR Abs that cross-react with self AChR, activate the complement cascade, and promote degradation of the postsynaptic membrane of the neuromuscular junction. In parallel, TAChR-specific T cells are induced, and their specific immunodominant epitope has been mapped to the sequence 97-116 of the AChR alpha subunit. A proliferative T cell response against the corresponding rat sequence (R97-116) was also found in TAChR-immunized rats. To test whether the rat (self) sequence can be pathogenic, we immunized Lewis rats with R97-116 or T97-116 peptides and evaluated clinical, neurophysiological, and immunological parameters. Clinical signs of the disease were noted only in R97-116-immunized animals and were confirmed by electrophysiological signs of impaired neuromuscular transmission. All animals produced Abs against the immunizing peptide, but anti-rat AChR Abs were observed only in animals immunized with the rat peptide. These findings suggested that EAMG in rats can be induced by a single peptide of the self AChR, that this sequence is recognized by T cells and Abs, and that breakdown of tolerance to a self epitope might be an initiating event in the pathogenesis of rat EAMG and MG. PMID- 14764746 TI - Adipocytokines and insulin resistance. PMID- 14764747 TI - Androgen excess in women: experience with over 1000 consecutive patients. AB - The objective of the present study was to estimate the prevalence of the different pathological conditions causing clinically evident androgen excess and to document the degree of long-term success of suppressive and/or antiandrogen hormonal therapy in a large consecutive population of patients. All patients presenting for evaluation of symptoms potentially related to androgen excess between October 1987 and June 2002 were evaluated, and the data were maintained prospectively in a computerized database. For the assessment of therapeutic response, a retrospective review of the medical chart was performed, after the exclusion of those patients seeking fertility therapy only, or with inadequate follow-up or poor compliance. A total of 1281 consecutive patients were seen during the study period. Excluded from analysis were 408 patients in whom we were unable to evaluate hormonal status, determine ovulatory status, or find any evidence of androgen excess. In the remaining population of 873 patients, the unbiased prevalence of androgen-secreting neoplasms was 0.2%, 21-hydroxylase deficient classic adrenal hyperplasia (CAH) was 0.6%, 21-hydroxylase-deficient nonclassic adrenal hyperplasia (NCAH) was 1.6%, hyperandrogenic insulin-resistant acanthosis nigricans (HAIRAN) syndrome was 3.1%, idiopathic hirsutism was 4.7%, and polycystic ovary syndrome (PCOS) was 82.0%. Fifty-nine (6.75%) patients had elevated androgen levels and hirsutism but normal ovulation. A total of 257 patients were included in the assessment of the response to hormonal therapy. The mean duration of follow-up was 33.5 months (range, 6-155). Hirsutism improved in 86%, menstrual dysfunction in 80%, acne in 81%, and hair loss in 33% of patients. The major side effects noted were irregular vaginal bleeding (16.1%), nausea (13.0%), and headaches (12.6%); only 36.6% of patients never complained of side effects. In this large study of consecutive patients presenting with clinically evident androgen excess, specific identifiable disorders (NCAH, CAH, HAIRAN syndrome, and androgen-secreting neoplasms) were observed in approximately 7% of subjects, whereas functional androgen excess, principally PCOS, was observed in the remainder. Hirsutism, menstrual dysfunction, or acne, but not alopecia, improved in the majority of patients treated with a combination suppressive therapy; although more than 60% experienced side effects. PMID- 14764748 TI - Role of the adipocyte, free fatty acids, and ectopic fat in pathogenesis of type 2 diabetes mellitus: peroxisomal proliferator-activated receptor agonists provide a rational therapeutic approach. PMID- 14764749 TI - Current approaches and recommended algorithm for the diagnostic localization of pheochromocytoma. PMID- 14764750 TI - Biochemical monitoring of disease activity after surgery for acromegaly. PMID- 14764751 TI - Significance of "abnormal" nadir growth hormone levels after oral glucose in postoperative patients with acromegaly in remission with normal insulin-like growth factor-I levels. AB - Our initial study in postoperative patients with acromegaly identified a group of patients in remission, as defined by normal IGF-I levels, but who had a subtle abnormality of GH suppression after oral glucose. To investigate the significance of this abnormality, we have undertaken further detailed testing of GH secretion and a longitudinal follow-up of some of these patients. Of the 110 postoperative patients with acromegaly evaluated by oral glucose tolerance test, 76 were in remission (i.e. normal IGF-I level), and of these subjects with acromegaly in remission, 50 had normal nadir GH (<0.14 microg/ml) (group I), and 26 had abnormal nadir GH (>0.14 microg/ml) (group II). Fourteen subjects in remission, seven from remission group I and seven from remission group II, underwent additional testing consisting of both hourly GH sampling over 8 h and, on a separate day, arginine stimulation testing. The mean of hourly GH was higher in group II (0.47 +/- 0.04 microg/liter) than in group I (0.19 +/- 0.07 microg/liter; P = 0.002). GH response to arginine was greater in group II than in group I (P < 0.01). Of those patients in remission from the initial cohort studied, 49 (30 subjects from group I and 19 from group II) underwent serial longitudinal oral glucose tolerance testing every 1-2 yr over a 1- to 6.5-yr period (mean follow-up, 3.2 yr). The initial pattern of GH suppression persisted in most patients. IGF-I levels remained normal in all patients in group II, but five subjects from group II developed an elevated IGF-I level and, thus, a biochemical recurrence. The rate of disease recurrence was greater in group II than in group I (P = 0.003). We have found that some postoperative subjects with acromegaly in remission with normal IGF-I levels have persistently abnormal nadir GH levels after oral glucose that may be accompanied by other evidence of greater GH secretion than postoperative patients with normal GH suppression. This abnormal pattern of GH suppression may be associated with increased risk of disease recurrence in some patients. PMID- 14764752 TI - Time for (more research on) testosterone. PMID- 14764753 TI - Exogenous testosterone or testosterone with finasteride increases bone mineral density in older men with low serum testosterone. AB - Older men, particularly those with low serum testosterone (T) levels, might benefit from T therapy to improve bone mineral density (BMD) and reduce fracture risk. Concerns exist, however, about the impact of T therapy on the prostate in older men. We hypothesized that the combination of T and finasteride (F), a 5 alpha-reductase inhibitor, might increase BMD in older men without adverse effects on the prostate. Seventy men aged 65 yr or older, with a serum T less than 12.1 nmol/liter on two occasions, were randomly assigned to receive one of three regimens for 36 months: T enanthate, 200 mg im every 2 wk with placebo pills daily (T-only); T enanthate, 200 mg every 2 wk with 5 mg F daily (T+F); or placebo injections and pills (placebo). Low BMD was not an inclusion criterion. We obtained serial measurements of BMD of the lumbar spine and hip by dual x-ray absorptiometry. Prostate-specific antigen (PSA) and prostate size were measured at baseline and during treatment to assess the impact of therapy on the prostate. Fifty men completed the 36-month protocol. By an intent-to-treat analysis including all men for as long as they contributed data, T therapy for 36 months increased BMD in these men at the lumbar spine [10.2 +/- 1.4% (mean percentage increase from baseline +/- SEM; T-only) and 9.3 +/- 1.4% (T+F) vs. 1.3 +/- 1.4% for placebo (P < 0.001)] and in the hip [2.7 +/- 0.7% (T-only) and 2.2 +/- 0.7% (T+F) vs. -0.2 +/- 0.7% for placebo, (P < or = 0.02)]. Significant increases in BMD were seen also in the intertrochanteric and trochanteric regions of the hip. After 6 months of therapy, urinary deoxypyridinoline (a bone-resorption marker) decreased significantly compared with baseline in both the T-only and T+F groups (P < 0.001) but was not significantly reduced compared with the placebo group. Over 36 months, PSA increased significantly from baseline in the T-only group (P < 0.001). Prostate volume increased in all groups during the 36-month treatment period, but this increase was significantly less in the T+F group compared with both the T-only and placebo groups (P = 0.02). These results demonstrate that T therapy in older men with low serum T increases vertebral and hip BMD over 36 months, both when administered alone and when combined with F. This finding suggests that dihydrotestosterone is not essential for the beneficial effects of T on BMD in men. In addition, the concomitant administration of F with T appears to attenuate the impact of T therapy on prostate size and PSA and might reduce the chance of benign prostatic hypertrophy or other prostate-related complications in older men on T therapy. These findings have important implications for the prevention and treatment of osteoporosis in older men with low T levels. PMID- 14764754 TI - The calcitonin conundrum--is it time for routine measurement of serum calcitonin in patients with thyroid nodules? PMID- 14764755 TI - Frequency and relevance of elevated calcitonin levels in patients with neoplastic and nonneoplastic thyroid disease and in healthy subjects. AB - Routine measurement of serum calcitonin (CT) has been recently proposed for all patients with neoplastic thyroid disease to detect clinically occult medullary thyroid carcinoma (MTC). Data on the prevalence of elevated CT levels in nonneoplastic thyroid disease or in healthy subjects have not been reported to date. Four hundred and fourteen consecutive patients with suspected thyroid disease and 362 healthy controls underwent thyroid examination with measurement of basal serum CT. Whenever serum CT was 10 pg/ml or more, a pentagastrin (PG) stimulation test was performed. Twenty-eight of 414 patients (6.8%) showed elevated basal serum CT levels, 15 of them with nonneoplastic thyroid disease, and the remaining 13 subjects with neoplastic thyroid disease. Four patients with abnormal PG testing (stimulated CT, > or = 100 pg/ml) were identified. Three of them had biochemical and sonographical evidence of thyroiditis. Elevated basal CT levels were significantly more frequent in patients with Hashimoto's thyroiditis (HT; P < 0.05). One female patient with HT had a 5-mm nodule, which was classified as MTC. None of the 6 out of 362 healthy controls with elevated basal CT (1.7%) presented an abnormal PG test. Our data suggest that basal CT measurements can be of use in the detection/screening of MTC not only in subjects with neoplastic thyroid disorders, but also in patients with immunological evidence of HT. They also confirm earlier reports on the essential value of PG stimulation testing, even when basal plasma CT levels are only modestly elevated, with regard to establishing the diagnosis of MTC or its premalignant associated conditions (micro-MTC and neoplastic C cell hyperplasia). PMID- 14764756 TI - Serum testosterone assays--accuracy matters. PMID- 14764757 TI - Measurement of free testosterone in normal women and women with androgen deficiency: comparison of methods. AB - Androgen deficiency in women is increasingly recognized as a new clinical syndrome and has raised our awareness of the importance of accurate and well validated measurements of serum free testosterone (T) concentrations in women. Therefore, we compared serum free T levels measured by equilibrium dialysis to those measured by a direct RIA (analog method) and to those calculated from the law of mass action (requires the measurement of total T and SHBG). We also calculated the free androgen index, 100 x T/SHBG, as a simple index known to correlate with free T. Subjects were 147 women with variable androgen and estrogen statuses. All were studied three times in 1 month and included women 1) with regular menses (estrogen positive, T positive), 2) more than 50 yr old and not receiving estrogen (estrogen negative, T positive), 3) receiving estrogen (estrogen positive, T negative), and 4) with severe androgen deficiency secondary to hypopituitarism (estrogen negative, T negative). Calculated values for free T using the laws of mass action correlated well with those obtained from equilibrium dialysis (r = 0.99; P < 0.0001). However, the agreement depended strongly on the specific assays used for total T and SHBG. In contrast, the direct RIA method had unacceptably high systematic bias and random variability and did not correlate as well with equilibrium dialysis values (r = 0.81; P < 0.0001). In addition, the lower limit of detection was higher for the direct RIA than for equilibrium dialysis or calculated free T. Free androgen index correlates well with free T by equilibrium dialysis (r = 0.93; P < 0.0001), but is a unitless number without reference to the physical reality of free T. We conclude that the mass action equation and equilibrium dialysis are the preferred methods for use in diagnosing androgen deficiency in women. PMID- 14764758 TI - Measurement of total serum testosterone in adult men: comparison of current laboratory methods versus liquid chromatography-tandem mass spectrometry. AB - The diagnosis of male hypogonadism requires the demonstration of a low serum testosterone (T) level. We examined serum T levels in pedigreed samples taken from 62 eugonadal and 60 hypogonadal males by four commonly used automated immunoassay instruments (Roche Elecsys, Bayer Centaur, Ortho Vitros ECi and DPC Immulite 2000) and two manual immunoassay methods (DPC-RIA, a coated tube commercial kit, and HUMC-RIA, a research laboratory assay) and compared results with measurements performed by liquid chromatography-tandem mass spectrometry (LC MSMS). Deming's regression analyses comparing each of the test results with LC MSMS showed slopes that were between 0.881 and 1.217. The interclass correlation coefficients were between 0.92 and 0.97 for all methods. Compared with the serum T concentrations measured by LC-MSMS, the DPC Immulite results were biased toward lower values (mean difference, -90 +/- 9 ng/dl) whereas the Bayer Centaur data were biased toward higher values (mean difference, +99 +/- 11 ng/dl) over a wide range of serum T levels. At low serum T concentrations (<100 ng/dl or 3.47 nmol/liter), HUMC-RIA overestimated serum T, Ortho Vitros ECi underestimated the serum T concentration, whereas the other two methods (DPC-RIA and Roche Elecsys) showed differences in both directions compared with LC-MSMS. Over 60% of the samples (with T levels within the adult male range) measured by most automated and manual methods were within +/- 20% of those reported by LC-MSMS. These immunoassays are capable of distinguishing eugonadal from hypogonadal males if adult male reference ranges have been established in each individual laboratory. The lack of precision and accuracy, together with bias of the immunoassay methods at low serum T concentrations, suggests that the current methods cannot be used to accurately measure T in females or serum from prepubertal subjects. PMID- 14764759 TI - Toward precise forecasting of autoimmune endocrinopathy. PMID- 14764760 TI - Activating antibodies to the calcium-sensing receptor in two patients with autoimmune hypoparathyroidism. AB - Autoimmune hypoparathyroidism is thought to result from immune-mediated destruction of the parathyroid glands. We encountered two patients with hypoparathyroidism and other autoimmune conditions (Graves' disease and Addison's disease, respectively) in whom autoimmune destruction of the parathyroid glands had not taken place. In the first, a histologically normal parathyroid gland was observed at the time of subtotal thyroidectomy; and in the second, the hypoparathyroidism remitted spontaneously. Both patients had antibodies that reacted with the cell surface of bovine parathyroid cells and human embryonic kidney (HEK293) cells transfected with the extracellular calcium-sensing receptor (CaR) but not with nontransfected HEK293 cells. The antibodies also reacted with the same bands on Western analysis of extracts of bovine parathyroid tissue and CaR-transfected HEK293 cells that were identified by an authentic, polyclonal, anti-CaR antiserum and reacted with several peptides with sequences from the CaR's extracellular domain. These anti-CaR antibodies activated the receptor based on their ability to increase inositol phosphate accumulation, activate MAPK, and inhibit PTH secretion. These results, therefore, demonstrate that patients with the biochemical findings of primary hypoparathyroidism can harbor activating antibodies to the CaR, which, in the two cases studied here, did not produce irreversible destruction of the parathyroid glands. PMID- 14764761 TI - Prevalence and clinical associations of 10 defined autoantibodies in autoimmune polyendocrine syndrome type I. AB - The prevalence of autoantibodies against nine intracellular enzyme autoantigens, namely 21-hydroxylase, side-chain cleavage enzyme (SCC), 17 alpha-hydroxylase, glutamic acid decarboxylase 65, aromatic L-amino acid decarboxylase, tyrosine phosphatase-like protein IA-2, tryptophan hydroxylase (TPH), tyrosine hydroxylase, cytochrome P450 1A2, and against the extracellular calcium-sensing receptor, was assessed in 90 patients with autoimmune polyendocrine syndrome type I. A multivariate logistic regression analysis was performed for the presence of autoantibodies as independent predictors for different disease manifestations. Reactivities against 21-hydroxylase and SCC were associated with Addison's disease with odds ratios (ORs) of 7.8 and 6.8, respectively. Hypogonadism was exclusively associated with autoantibodies against SCC with an OR of 12.5. Autoantibodies against tyrosine phosphatase-like protein IA-2 were associated with insulin-dependent diabetes mellitus with an OR of 14.9, but with low sensitivity. Reactivities against TPH and, surprisingly, glutamic acid decarboxylase 65, were associated with intestinal dysfunction, with ORs of 3.9 and 6.7, respectively. TPH reactivity was the best predictor for autoimmune hepatitis, with an OR of 27.0. Hypoparathyroidism was not associated with reactivity against any of the autoantigens tested. No reactivity against the calcium-sensing receptor was found. Analysis of autoantibodies in autoimmune polyendocrine syndrome type I patients is a useful tool for establishing autoimmune manifestations of the disease as well as providing diagnosis in patients with suspected disease. PMID- 14764762 TI - Glucocorticoid action networks--an introduction to systems biology. PMID- 14764763 TI - Common polymorphisms in the glucocorticoid receptor gene are associated with adrenocortical responses to psychosocial stress. AB - Chronic dysregulation of hypothalamus-pituitary-adrenal axis activity is related to several stress-related disorders. Evidence suggests that polymorphisms in the glucocorticoid receptor (GR) gene may have an impact on this neuroendocrine system. In the present investigation, 112 healthy males were studied to estimate the impact of three GR gene polymorphisms (BclI RFLP, N363S, ER22/23EK) on cortisol and ACTH responses to psychosocial stress (Trier Social Stress Test) and pharmacological stimulation (1 microg ACTH(1-24), 0.5 mg dexamethasone). Because only four ER22/23EK heterozygotes were identified, these subjects were not statistically analyzed. Compared with subjects with the wild-type GR genotype (n = 36), 363S allele carriers (n = 10) showed significantly increased salivary cortisol responses to stress, whereas the BclI genotype GG (n = 18) was associated with a diminished cortisol response. BclI heterozygotes and homozygotes (GG) exhibited a trend toward lower ACTH responses, compared with wild-type subjects and 363S carriers. The cortisol response to ACTH(1-24) administration was not significantly different between genotypes. After dexamethasone ingestion, 363S carriers showed a trend toward an enhanced cortisol suppression. This is the first report documenting an impact of GR gene polymorphisms on cortisol (and perhaps ACTH) responses to psychosocial stress. These variants may contribute to the individual vulnerability for hypothalamus pituitary-adrenal-related disorders. PMID- 14764764 TI - Tumors metastatic to the pituitary gland: case report and literature review. AB - Tumors metastatic to the pituitary gland are an unusual complication of systemic cancer typically seen in elderly patients with diffuse malignant disease. Breast and lung are the commonest sites of the primary tumor, whereas diabetes insipidus is the most frequent symptom at presentation. Their rarity and usually indolent course, as well as the lack of specific clinical and radiological features, impede their differentiation from other more common sellar area lesions, particularly when history of malignancy is absent. Management of these patients may also be very difficult because the prognosis depends on the course of the primary neoplasm. A 68-yr-old man, with no history of malignancy, presented with recent onset of hypopituitarism, mild diabetes insipidus, headaches, left oculomotor nerve palsy, and progressive bilateral deterioration of visual acuity and visual fields. Magnetic resonance imaging revealed a large sellar mass compressing the optic chiasm and invading the left cavernous sinus, whereas a prolactin elevation at 438.6 ng/ml (19.73 nmol/liter) was noted. Decompression of the sellar region was attempted, and pathology disclosed a metastatic hepatocellular carcinoma. On postoperative investigation, primary liver tumor was identified and confirmed by biopsy. The patient improved transiently but died 3 months after diagnosis because of deterioration of the liver disease. The relevant literature is reviewed in light of this unusual case, illustrating the problems in the diagnosis and management of patients with metastasis to the pituitary. PMID- 14764765 TI - A case of dopamine agonists inhibiting pancreatic polypeptide secretion from an islet cell tumor. AB - A patient with a large prolactinoma developed a metastatic islet cell tumor secreting pancreatic polypeptide. Dopamine agonist drugs reduced the prolactin levels to normal, caused a 7-fold decrease in the pancreatic polypeptide levels, and inhibited the liver metastases. Elevated chromogranin A levels also normalized on the higher doses of bromocriptine. Dopamine receptors are found in many endocrine tissues, and the expression of dopamine-2 receptor on endocrine tumors establishes the potential for response to dopamine agonist treatment. The relatively benign risk profile of dopaminergic agents makes further testing of these drugs to treat neuroendocrine tumors a worthwhile endeavor. PMID- 14764766 TI - Two cases of thyroid carcinoma that were not stimulated by recombinant human thyrotropin. AB - Recombinant human TSH (rhTSH) is being widely used to monitor patients who were previously treated for differentiated thyroid cancers for evidence of recurrence. Its value lies in the avoidance of recurrent episodes of hypothyroidism in the follow-up protocols. rhTSH is also being evaluated as a potential therapeutic adjunct that would spare patients the experience of becoming hypothyroid when undergoing thyroid remnant ablation or treatment for metastases. In some centers, rhTSH is also used to support compassionate care of patients with advanced disease who cannot safely become hypothyroid. The (131)I uptake response to rhTSH, presently an off-label application, is expected to be similar to that of endogenously raised TSH levels. The two cases presented here are cautionary tales in which (131)I uptake by metastases was present under hypothyroid conditions, but absent in one patient and present in only a portion of the lesions in the other, with rhTSH stimulation. PMID- 14764767 TI - Patients with classic congenital adrenal hyperplasia have decreased epinephrine reserve and defective glucose elevation in response to high-intensity exercise. AB - Classic congenital hyperplasia (CAH) is characterized by impaired adrenocortical function with a decrease in cortisol and aldosterone secretion and an increase in androgen secretion. Adrenomedullary function is also compromised due to developmental defects in the formation of the adrenal medulla, leading to decreased production of epinephrine. To examine the response to a natural stressful stimulus in patients with classic CAH, we studied hormonal, metabolic, and cardiorespiratory parameters in response to a standardized high-intensity exercise protocol in nine adolescent patients with CAH and nine healthy controls matched for gender, age, and percent body fat. The same relative workload was applied, based on individual maximal aerobic capacity, and all patients received their usual glucocorticoid and mineralocorticoid replacement. When compared with their normal counterparts, patients with CAH had significantly lower epinephrine levels both at baseline and at peak exercise (P < 0.01), whereas norepinephrine levels did not differ. Blood glucose concentrations were similar at baseline, but the normal exercise-induced rise observed in the healthy controls was significantly blunted in the CAH patients (P < 0.01). Peak heart rate was also lower in CAH patients than healthy controls (P < 0.05). As expected, the normal exercise-induced increase in cortisol was not observed in patients with CAH. No significant differences were found in serum levels of insulin, glucagon, GH, lactate and free fatty acids, blood pressure, or ability to sustain exercise between the two groups. Patients with CAH replaced with glucocorticoids have decreased adrenomedullary reserve and impaired exercise-induced changes in glucose but normal short-term high-intensity exercise performance. Whether the combination of epinephrine and cortisol deficiency poses a risk for hypoglycemia and/or decreased endurance during long-term physical stress has to be determined. PMID- 14764768 TI - Acromegalic axial arthropathy: a clinical case-control study. AB - Arthropathy is the major cause of morbidity in acromegaly. To feature the spinal involvement, 54 patients with active acromegaly (27 men, 27 women; age range, 21 69 yr) and 54 sex-, age-, and body mass index-matched healthy controls were enrolled in this observational analytical prospective case-control study. A questionnaire to describe onset, duration, and severity of articular symptoms; rheumatological examination, including vertebral and chest mobility, Schober test, thorax expansion, and axial radiological study; and IGF-I, GH, insulin, and glucose level measurement (baseline and after an oral glucose tolerance test) was used to investigate the prevalence of arthropathy and correlate these findings with hormonal parameters. Axial arthropathy was found in 28 patients (52%) and 12 controls (22%; chi(2) = 8.9; P = 0.003). In detail, spinal mobility was reduced in 30 patients (56%) and 10 controls (18%; chi(2) = 14.3; P < 0.0001), thoracic cage was involved in six patients (11%), alterations of spinal profile were observed in 37 patients (68%) and 15 controls (28%; chi(2) = 16.3; P < 0.0001), and increased L2 vertebra diameters were observed in 34 patients (63%) and none of the controls (chi(2) = 46.7; P < 0.0001). Narrowing and widening of L2-L3 disk space were found in 20 (37%) and seven (13%) patients, respectively. Features of diffuse idiopathic skeletal hyperostosis (DISH) were found in 11 patients (20%) and none of the controls (chi(2) = 10.1; P < 0.001). Disease duration was correlated with vertebral body height (P = 0.001) or intervertebral space height (P = 0.02), and lumbar mobility with thorax expansion (P = 0.004); DISH severity was correlated with basal (P = 0.04) and peak (P = 0.01) glucose levels after glucose load. In conclusion, chronic GH and IGF-I excess typically affects the axial skeleton with development of severe alterations of spine morphology and function until features of DISH occur. An early diagnosis of acromegaly is mandatory to reduce the severity of spine abnormalities as they were significantly higher in patients with longer disease duration. PMID- 14764769 TI - Incidence of growth hormone deficiency in pediatric-onset Langerhans cell histiocytosis: efficacy and safety of growth hormone treatment. AB - We retrospectively studied 61 patients with GH deficiency (GHD), identified among 589 patients with Langerhans cell histiocytosis (LCH) enrolled in a nationwide survey between 1993 and 2001. Overall, 141 patients in the survey developed diabetes insipidus. The median follow-up of the 61 patients with GHD was 12 yr. The 5- and 10-yr risks of GHD among patients with diabetes insipidus were 34.7 +/ 4.5% and 53.7 +/- 5.2%, respectively. Growth velocity decreased soon after LCH diagnosis in patients who developed GHD, and anterior pituitary height, estimated by magnetic resonance imaging, was significantly reduced relative to patients who remained free of GHD. GH replacement therapy was administered to 47 of the 61 patients with GHD. Among GH-treated patients, median final height (-0.8 SD) was significantly greater than median height at GHD diagnosis (-1.6 SD) but remained below midparental (target) height. Among patients with pituitary involvement, the number of LCH disease episodes appeared not significantly influenced by GHD or GH administration, suggesting an absence of deleterious effect of GH therapy on LCH disease activity. PMID- 14764770 TI - Cognitive and motor development of children with and without congenital adrenal hyperplasia after early-prenatal dexamethasone. AB - Dexamethasone (DEX) administration to the pregnant woman has become the treatment of choice for the prevention of genital masculinization in female fetuses affected with congenital adrenal hyperplasia (CAH). Although no somatic teratological side effects have been found to date, recent animal research has shown adverse effects of glucocorticoids on brain structures such as the hippocampus, raising concerns about possible functional side effects of DEX on human development. The current survey of 487 children, 1 month to 12 yr of age, focused on cognitive and motor development. The mothers of 174 prenatally DEX exposed children (including 48 with CAH) and 313 unexposed children (including 195 with CAH) completed four standardized developmental questionnaires about their children. None of the comparisons of prenatally DEX-exposed children and unexposed controls was significant. Among the DEX-exposed children, increased duration of DEX exposure was correlated with significantly fewer developmental delays on three variables of one of the questionnaires, but none of the correlations reached significance, when Bonferroni corrections for multiple correlations were used. With the methods used, we were unable to document any adverse effects of early-prenatal DEX treatment in the doses recommended for the treatment of pregnancies at risk for CAH on motor and cognitive development. PMID- 14764771 TI - Metabolic and endocrine effects of a polyunsaturated fatty acid-rich diet in polycystic ovary syndrome. AB - Effects of a polyunsaturated fatty acid (PUFA)-rich diet were investigated in 17 polycystic ovary syndrome (PCOS) patients. After a 3-month habitual diet period, dietary fats were partly replaced with PUFAs for another 3 months. The PUFA-rich diet increased plasma linoleic acid from 28.36 +/- 1.00% to 33.76 +/- 1.08% (P < 0.002) and alpha-linolenic acid from 0.52 +/- 0.03% to 1.06 +/- 0.10% (P < 0.0001). Fasting glucose increased from 76 +/- 3 to 95 +/- 3 mg/dl (4.2 +/- 0.2 to 5.30.2 mmol/liter; P < 0.0001), and the area under the curve for glucose during oral glucose tolerance test increased from 421 +/- 34 to 503 +/- 31 mg/dl (23.4 +/- 1.9 to 27.9 +/- 1.7 mmol/liter; P < 0.001). Plasma insulin did not change either at fasting or during oral glucose tolerance test. Fasting plasma free fatty acids decreased from 0.596 +/- 0.048 to 0.445 +/- 0.058 mg/dl (P = 0.037), and ketone bodies decreased from 9.14 +/- 1.57 to 3.63 +/- 0.62 mg/dl (895 +/- 154 to 356 +/- 61 micromol/liter; P < 0.003). Plasma 15 deoxyprostaglandin J(2) tended to decrease (from 239 +/- 65 to 171 +/- 60 ng/ml; P = 0.053). Plasma testosterone, free testosterone, SHBG, dehydroepiandrosterone sulfate, LH, FSH, and urinary estrogen conjugates did not change. Urinary pregnanediol 3-glucuronide increased from 18.6 +/- 2.2 to 31.0 +/- 5.7 micro g/mg creatinine (P = 0.038). In conclusion, increased dietary PUFA intake can exert significant metabolic and endocrine effects in women with PCOS. PMID- 14764772 TI - Six months of treatment with cabergoline restores sexual potency in hyperprolactinemic males: an open longitudinal study monitoring nocturnal penile tumescence. AB - This open longitudinal study investigated the prevalence of depressed sexual potency by monitoring erectile dysfunction using nocturnal penile tumescence (NPT) in 51 consecutive men with hyperprolactinemia (41 macroprolactinomas and 10 microprolactinomas) and evaluated potential reversibility of sexual failure after 6 months of treatment with cabergoline. Fifty-one healthy men served as controls. Compared with controls, the patients with either micro- or macroprolactinoma had low testosterone levels with severe alterations of erectile function. Testosterone deficiency was present in 73.2% of macro- and 50% of microprolactinomas; reduced libido and sexual potency were referred by 53.6% of macroprolactinomas, 50% of microprolactinomas, and none of controls. Fewer than three erectile events per night by NPT were found in 96.7% of patients and 13.7% of controls (P < 0.0001). After 6 months of cabergoline treatment, prolactin levels normalized in 74.5% of patients: 73.2% of macroprolactinomas and 80% of microprolactinomas. Testosterone levels normalized in 68.6% of patients, whereas NPT normalized in 60.6% of patients who had normalized prolactin levels and in 7.7% of patients who did not. In conclusion, at study entry, 50% of the patients complained of sexual disturbances, 96.7% of whom had an impairment of erectile events per night compared with 13.7% of controls. Six months of treatment with cabergoline normalized testosterone levels in most cases, thus restoring and maintaining during treatment the capability of normal sexual activity in hyperprolactinemic males. PMID- 14764773 TI - Effects of alendronate and hormone replacement therapy, alone and in combination, on bone mass and markers of bone turnover in elderly women with osteoporosis. AB - The aim of the study was to compare alendronate, hormone replacement therapy (HRT), and their combination in treatment of osteoporosis in elderly postmenopausal women. Ninety patients, aged 65-80 yr (mean 71), with a T-score of bone mineral density (BMD) of 2.5 or less at either the lumbar spine or the femoral neck were randomized to receive daily 10 mg alendronate (n = 30), 2 mg estradiol plus 1 mg norethisterone acetate (n = 30) (HRT), or their combination (n = 30) for 2 yr. BMD of the lumbar spine and the upper femur was measured at baseline and after 1 and 2 yr of treatment. Urinary excretion of type I collagen aminoterminal telopeptide as related to creatinine and serum type I procollagen aminoterminal propeptide was assayed at baseline and at 6-month intervals thereafter. Increases of 9.1-11.2% in lumbar spine BMD at 2 yr were similar in the study groups. Only HRT increased femoral neck BMD statistically significantly (P < 0.0001 for a change from baseline) at both 1 (+4.9%; P =NS vs. the other groups) and 2 yr (+5.8%; P < 0.05 vs. the other groups). Total hip BMD increased similarly in all study groups. Percentage reductions in urinary type I collagen aminoterminal telopeptide in the HRT group (60.2-62.7%) were significantly smaller than those in the combination group (78.1-80.4%) (P < 0.0001-0.0069) and the alendronate-only group (72.4-76.1%) (P = 0.047 at 24 months). Serum type I procollagen aminoterminal propeptide decreased less in the HRT group (53.6-59.8%) than in the other groups [73.0-75.0% in the alendronate group (P < 0.001 at 12 months); 67.0-71.5% in the combination group (P < 0.0001 at 12 months, P = 0.013 at 24 months)]. We conclude that in elderly postmenopausal women with osteoporosis, the combination of HRT and alendronate did not offer an extra gain of bone mass over either treatment alone. In terms of BMD changes, the single treatments were equally effective, but the reductions in bone markers were less with HRT than with alendronate. PMID- 14764774 TI - Effect of calcium supplementation on weight and fat loss in women. AB - Data suggest that a diet deficient in calcium is associated with higher body weight and that augmenting calcium intake may reduce weight and fat gain or enhance loss. Our aim was to determine whether calcium supplementation during a weight loss intervention affects body fat or weight loss. Data were combined from three separate 25-wk randomized, double blind, placebo-controlled trials of 1000 mg/d calcium supplementation in 100 premenopausal and postmenopausal women. The primary outcome measures were change in body weight and fat mass adjusted for baseline values. There were no significant differences in body weight or fat mass change between the placebo and the calcium-supplemented groups in the pooled analysis (adjusted mean +/- SE; body weight, placebo -6.2 +/- 0.7 vs. Ca -7.0 +/- 0.7 kg; fat mass, placebo -4.5 +/- 0.6 vs. Ca -5.5 +/- 0.6 kg), and no significant interactions of calcium supplementation with menopausal/diet status. Analysis as separate trials also found no significant differences between the placebo and the calcium groups. Calcium supplementation did not significantly affect amount of weight or fat lost by women counseled to follow a moderately restricted diet for 25 wk. Nevertheless, the magnitude and direction of the differences for group means are consistent with a hypothesized small effect. PMID- 14764775 TI - A single-dose comparison of the acute effects between the new somatostatin analog SOM230 and octreotide in acromegalic patients. AB - Treatment with the somatostatin receptor (sst) subtype 2 predominant analogs octreotide and lanreotide induces clinical and biochemical cure in approximately 65% of acromegalic patients. GH-secreting pituitary adenomas, which are not controlled, also express sst(5). We compared the acute effects of octreotide and SOM230, a new somatostatin analog with high affinity for sst(1,2,3,5) on hormone release in acromegalic patients. In a single-dose, proof-of-concept study, 100 microg octreotide and 100 and 250 microg SOM230 were given s.c. to 12 patients with active acromegaly. Doses of 100 and 250 microg SOM230 dose-dependently suppressed GH levels from 2-8 h after administration (-38 +/- 7.7 vs. -61 +/- 6.7%, respectively; P < 0.01). A comparable suppression of GH levels by octreotide and 250 microg SOM230 was observed in eight patients (-65 +/- 7 vs. 72 +/- 7%, respectively). In three patients, the acute GH-lowering effect of 250 microg SOM230 was significantly superior to that of octreotide (-70 +/- 2 vs. -17 +/- 15%, respectively; P < 0.01). In one patient, the GH-lowering effect of octreotide was better than that of SOM230. Tolerability for SOM230 was good. Glucose levels were initially slightly elevated after octreotide and SOM230, compared with control day, whereas insulin levels were only significantly suppressed by octreotide. We conclude that SOM230 is an effective GH-lowering drug in acromegalic patients with the potential to increase the number of patients controlled during long-term medical treatment. PMID- 14764776 TI - Two distinct compound heterozygous constellations (R277X/IVS34-1G>C and R277X/R1511X) in the thyroglobulin (TG) gene in affected individuals of a Brazilian kindred with congenital goiter and defective TG synthesis. AB - In this study, we have extended our initial molecular studies of a nonconsanguineous family with two affected siblings and one of their nephews with congenital goiter, hypothyroidism, and marked impairment of thyroglobulin synthesis. Genomic DNA sequencing revealed that the index patient (affected nephew) was heterozygous for a single base change of a cytosine to a thymine at nucleotide 886 in exon 7 (886C>T, mother's mutation) in one allele and for a novel guanine to cytosine transversion at position -1 of the splice acceptor site in intron 34 (IVS34-1G>C, father's mutation) in the other allele. The two affected siblings inherited the 886C>T mutation from their mother and a previously reported cytosine to thymine transition at nucleotide 4588 in exon 22 from their father (4588C>T). The 886C>T and 4588C>T substitutions resulted in premature stop codons at amino acids 277 (R277X) and 1511 (R1511X), respectively. In vitro transcription analysis showed that the exon 35 is skipped entirely when the IVS34-1G>C mutation is present, whereas the wild-type allele is correctly spliced. SSCP (exon 7 and 35) and restriction analysis (exon 22) using Taq I indicated that the two affected siblings, the affected nephew, his mother, and his unaffected brother were all heterozygous for the R277X mutation. The two affected siblings, their father, and three unaffected siblings were all heterozygous for the R1511X mutation, whereas the affected nephew and his father were heterozygous for the IVS34-1G>C mutation. Moreover, in this kindred, we have characterized polymorphisms (insertion/deletion, microsatellite, and single nucleotide polymorphism) located within introns 18 and 29 and exon 44 that are associated with the described mutations. Haplotype analysis with these polymorphic markers in two unrelated Brazilian families (present family studied and previously reported family) harboring the R277X mutation suggests a founder effect for the R277X mutation. In conclusion, the affected individuals of this family are either compound heterozygous for R277X/IVS34-1G>C or R277X/R1511X. This observation further supports that thyroglobulin gene mutations display significant intraallelic heterogeneity. PMID- 14764777 TI - Long-term biochemical status and disease-related morbidity in 53 postoperative patients with acromegaly. AB - Assessment of postoperative disease activity of acromegaly is a major challenge. The consensus criteria for cure, which are glucose-suppressed GH less than 1 micro g/liter and normal IGF-I levels, might be discrepant, and their respective relationship to acromegaly-related morbidity is not well known. THE AIMS OF OUR STUDY WERE: firstly, to correlate plasma IGF-I with plasma glucose-suppressed GH concentrations; and secondly, to correlate each of these biochemical parameters with morbidity [impaired glucose tolerance (IGT), diabetes, and hypertension] in postoperative patients with acromegaly. Fifty-three patients with long-term follow-up (mean, 12.7 yr; range, 1-30 yr) after transsphenoidal surgery for acromegaly and 20 healthy subjects matched for age, sex, and body mass index were evaluated for plasma glucose [by 75-g oral glucose tolerance test (OGTT)], GH (by immunoradiometric assay), plasma IGF-I (by immunoradiometric assay), and blood pressure (BP) measurements. Remission was defined by a normal IGF-I. We identified 34 acromegalics in remission and 19 with active disease. There was no statistical difference between all three groups for age, sex, BMI, and for fasting and 2-h post-OGTT plasma glucose. The time elapsed since surgery was similar in both groups of acromegalics. The OGTT-GH nadir was less than 1 micro g/liter in 31 patients in remission (91.2%) and in nine patients with active disease (47.4%). Prevalence of IGT was lower in acromegalics in remission (14.7%) in comparison with patients with active disease (47.4%; P = 0.01). Plasma IGF-I and GH nadir cut-off of 0.25 microg/liter were strong predictors of abnormal glucose tolerance (odds ratio, 13.6; confidence interval, 2.5-73.7; P = 0.003). GH nadir cut-off of 1 microg/liter and basal GH of 2.5 microg/liter failed to predict abnormal glucose tolerance. There was no statistical difference for prevalence of hypertension and systolic BP values, but diastolic BP was significantly lower in patients in remission than in those with active disease (P < 0.05). Our observations indicate that the validity of the GH threshold of 1 microg/liter post OGTT might be inadequate as a criterion of biochemical remission of acromegaly and as a marker of associated comorbidities. However, normalized IGF-I concentrations and a lower GH cut-off value less than 0.25 micro g/liter are strongly associated with a lower prevalence of IGT and lower diastolic BP in long-term postoperative acromegaly. PMID- 14764778 TI - Reassessment of growth hormone status is required at final height in children treated with growth hormone replacement after radiation therapy. AB - The most appropriate way to manage GH replacement in the transition period to adulthood in children treated with GH for GH deficiency (GHD) is controversial. The Growth Hormone Research Society suggests that the retesting of GH status at final height (FH) is unnecessary in the presence of severe organic GHD, and cranial irradiation falls into this etiological category. This recommendation has never been validated. To investigate whether patients diagnosed in childhood as GHD secondary to irradiation require retesting after FH, GH status has been reassessed in a large cohort of irradiated children treated with GH during childhood. Seventy-three children underwent biochemical assessment of GH status after irradiation and again at FH after GH therapy had been discontinued; 66 and 67 of the 73 patients underwent two provocative tests at the two time points, respectively. The characteristics of the cohort include a median age at irradiation of 5 yr (range, 1-11 yr), a median biological effective dose (BED) of irradiation to the hypothalamic pituitary axis of 54 Gy (range, 23-82 Gy), and a median time of GH status reassessment after FH of 0.4 yr (range, 0-8.4 yr). During childhood, patients with all degrees of GHD (peak GH responses to provocative test < 6.7 ng/ml) are treated, whereas in adulthood, only patients with severe GHD (peak GH responses to provocative test < 3 ng/ml) are considered for GH replacement. GH status has been grouped as follows: group 1, peak GH less than 3 ng/ml to both tests (severe GHD); group 2, one test with a peak GH less than 3 ng/ml and the other test with a peak of 3 ng/ml or greater; group 3, peak GH of 3-6.7 ng/ml to both tests; group 4, one test with a peak GH of 3-6.7 ng/ml and the other test with a peak of more than 6.7 ng/ml; and group 5, peak GH more than 6.7 ng/ml to both tests (normal GH status). In childhood, the number of patients in groups 1, 2, 3, and 4 were 33, 22, 17, and one, respectively. At retesting, severe GHD was diagnosed in 21 (64%) of 33 patients who were diagnosed in childhood with severe GHD (group 1) and 17 (44%) of 39 patients who were diagnosed in childhood with moderate GHD (groups 2 and 3). In total, 35 (48%) of 73 patients in the whole cohort and 12 (36%) of 33 patients with severe GHD in childhood did not fulfill the severe GHD biochemical criteria for GH replacement in adulthood. Using multiple linear regression, GH status at retesting is predicted by BED, age at irradiation, and use of chemotherapy. In conclusion, the diagnosis of severe GHD in childhood secondary to irradiation should not be taken as irrefutable evidence of permanent severe organic GHD, and our recommendation is that retesting of GH status at FH should be mandatory. PMID- 14764779 TI - Factors influencing mortality in acromegaly. AB - Studies of acromegaly have shown a doubling of mortality compared with the general population. With the development of new modalities of treatment, it has become important to identify prognostic factors relating to mortality. Between 1964 and 2000, 208 acromegalic patients were followed for a mean of 13 yr at Auckland Hospital. Treatment was by surgery or radionuclide pituitary implantation, and all except 27 patients received pituitary radiation. Over the duration of the study, 72 patients died at a mean age of 61 +/- 12.8 yr. Those dying were significantly older at diagnosis, had a higher prevalence of hypertension and diabetes, and were more likely to have hypopituitarism. The observed to expected mortality ratio (O/E ratio) fell from 2.6 (95% confidence interval, 1.9-3.6) in those with last follow-up GH greater than 5 microg/liter to 2.5 (1.6-3.8), 1.6 (0.9-3), and 1.1 (0.5-2.1) for those with GH less than 5, less than 2, and less than 1 microg/liter, respectively (P < 0.001). Serum IGF-I, expressed as an SD score, was significantly associated with mortality, with O/E mortality ratios of 3.5 (95% confidence interval, 2.8-4.2) for those with an SD score greater than 2, 1.6 (0.6-2.6) for those with an SD score less than 2 (normal or low levels), and 1.0 (0.1-3) for those with an SD score less than zero. When assessed by multivariate analysis, last serum GH (P < 0.001), age, duration of symptoms before diagnosis (P < 0.03), and hypertension (P < 0.04) were independent predictors of survival. If IGF-I was substituted for GH, then survival was independently related to last IGF-I SD score (P < 0.02), indicating that GH and IGF-I act equivalently as predictors of mortality. These findings indicate that reduction of GH to less than 1 microg/liter or normalization of serum IGF-I reduces mortality to expected levels. PMID- 14764780 TI - Adrenal sensitivity to adrenocorticotropin 1-24 is reduced in patients with autoimmune polyglandular syndrome. AB - Autoimmune polyglandular syndromes are fairly common diseases that are classified into four constellations based on the clinical clustering of the various component diseases. In types 1, 2, and 4, primary adrenal insufficiency due to an autoimmune process is usually present, but its diagnosis is often delayed because it is difficult to detect in a subclinical phase. It is widely accepted that the classical dose of 250 microg ACTH(1-24) is supramaximal, whereas 0.06 microg has been shown to be one of the lowest ACTH doses that is able to stimulate adrenal secretion in normal young subjects. The aim of this study was to clarify the sensitivity and maximal secretory response of the adrenal gland to ACTH in a group of patients with at least two autoimmune diseases, without clinical signs and symptoms of overt or subclinical hypocortisolism. Cortisol (F), aldosterone (A), and dehydroepiandrosterone (DHEA) responses to the sequential administration of very low and supramaximal ACTH(1-24) doses [0.06 microg followed by 250 microg ACTH(1-24) i.v. at 0 and +60 min] were studied in 18 patients with at least two autoimmune diseases (AP; age, 20-40 yr; body mass index, 22-26 kg/m(2)). The results in the patients were compared with the results recorded in 12 normal age matched control subjects (CS; age, 22-34 yr; body mass index, 20-25 kg/m(2)). At baseline, ACTH levels in AP were within the normal range but higher (P < 0.05) than in CS, whereas F, A, DHEA, urinary-free F, and plasma renin activity were similar in both groups. F, A, and DHEA responses to ACTH were dose dependent in both groups. However, in AP, F, A, and DHEA levels showed no response to the 0.06 micro g ACTH dose, which, in turn, elicited clear responses (P < 0.01) in CS. On the other hand, F, A, and DHEA responses to 250 microg ACTH in AP were not different from those in CS. In conclusion, patients with autoimmune diseases who displayed a normal basal adrenal function showed a loss of F, A, and DHEA response to the very low ACTH dose, although they were normal responders to the high ACTH dose. These data are likely to indicate that a reduced sensitivity to ACTH in all adrenal zones occurs in patients with different types of autoimmune disease. PMID- 14764782 TI - The effect of a six-month exercise program on very low-density lipoprotein apolipoprotein B secretion in type 2 diabetes. AB - The dyslipidemia and insulin resistance of type 2 diabetes can be improved by aerobic exercise. The effect of 6 months supervised exercise on very low-density lipoprotein (VLDL) apolipoprotein B metabolism was investigated in patients with type 2 diabetes. Moderately obese patients (n = 18) were randomized into supervised (n = 9) and unsupervised (n = 9) exercise groups. All patients were given a training session and a personal exercise program and asked to exercise four times per week at 70% maximal oxygen uptake for 6 months. Patients in the supervised group had a weekly session with an exercise trainer. VLDL apolipoprotein (apo)B metabolism was measured with an infusion of 1-(13)C leucine before and after 6 months of the exercise program. Supervised exercise for 6 months resulted in a significant within-group decrease in percent hemoglobin A1c (P < 0.001), body fat (P < 0.004), nonesterified fatty acid (P < 0.04), and triglycerides (P < 0.05) and an increase in insulin sensitivity (P < 0.01). There was a decrease in VLDL apoB pool size (160.8 +/- 42.6 to 84.9 +/- 23.2 mg, P < 0.01) and VLDL apoB secretion rate (11.3 +/- 2.6 to 5.5 +/- 2.0 mg/kg.d, P < 0.05) with no change in fractional catabolic rate. In a between-group comparison, the decrease in VLDL apoB secretion rate in the supervised group did not achieve significance. This study demonstrates that in type 2 diabetes, a supervised exercise program reduces VLDL apoB pool size, which may be due to a decrease in VLDL apoB secretion rate. PMID- 14764781 TI - Transdermal testosterone application: pharmacokinetics and effects on pubertal status, short-term growth, and bone turnover. AB - The aim of the study was to assess the effect of transdermal testosterone on free testosterone concentrations in saliva and on short-term growth and bone turnover in boys with growth or pubertal delay. A prospective, randomized, crossover study was conducted over 26 wk with 4 wk of run-in, 8 wk of treatment I (8 or 12 h), 4 wk of washout, 8 wk of treatment II (8 or 12 h), and 4 wk of final washout. The main outcome measures were salivary testosterone profiles during the different study periods; weekly change in lower leg length (LLL) as measured by knemometry, i.e. LLL velocity; absolute and percentage change in bone alkaline phosphatase (bALP) levels; and deoxypyridinoline cross-links measured in urine. Eight boys who took part in the study had a median age of 13.5 yr (range, 12.4-14.9 yr), testicular volume of 3 ml (range, 2-6 ml), height SD score of -2.4 (range, -1.44 to -3.35), and bone age delay of 2 yr (range, 1-3.2 yr). Median salivary testosterone during 8- and 12-h treatments [179 pg/ml (range, 7-3579 pg/ml) and 150 pg/ml (range, 12-3472 pg/ml) (not significant)] was significantly higher than during the run-in and washout blocks (P < 0.0001) [9 pg/ml (range, <7 to 122 pg/ml) and 13 pg/ml (range, <7 to 285 pg/ml) (not significant)]. LLL velocity in the treatment blocks (median, 0.64 mm/wk; range, 0.1-1.08 mm/wk) was significantly higher than during the run-in and washout periods (median, 0.48 mm/wk; range, -0.06 to 0.92 mm/wk) (P < 0.001). The main rise in bALP occurred during the first treatment block with a median percentage change in bALP of 44.2% (range, -4 to 87%) and a smaller percentage change in bALP at the end of the second treatment block of 9.8% (range, -4 to 55%). The increases in bALP were not significantly different between the 8- and 12-h treatment periods, and there was no significant decline during the washout periods. Overnight transdermal testosterone application, as Virormone (5 mg), may be a potentially acceptable method of induction of puberty and stimulates short-term growth and bone turnover. PMID- 14764783 TI - Low-dose recombinant human growth hormone as adjuvant therapy to lifestyle modifications in the management of obesity. AB - Obese individuals are in a reduced GH/IGF-I state that may be maladaptive. Fifty nine obese men and premenopausal menstruating women (body mass index, 36.9 +/- 5.0 kg/m(2)) were randomized to a double-blind, placebo-controlled trial of low dose recombinant human GH (rhGH). During the 6-month intervention, subjects self administered daily rhGH or equivalent volume of placebo at 200 micro g (1.9 +/- 0.3 microg/kg for men, 2.0 +/- 0.3 microg/kg for women); after 1 month, the dose was increased to 400 microg (3.8 +/- 0.5 microg/kg) in men and 600 microg (6.0 +/ 0.8 microg/kg) in women. rhGH was then discontinued, and subjects were followed up after 3 months. Forty completed the intervention, and 39 completed the follow up. Drop-out rates between rhGH vs. placebo groups were not different (chi(2) = 1.45; P = 0.228). One subject discontinued the drug due to an rhGH-related side effect. Body weight (BW) decreased with rhGH from 100.4 +/- 13.2 to 98.0 +/- 15.6 kg at 6 months (P = 0.04) and was sustained at 98.1 +/- 16.6 kg at 9 months (P = 0.02). BW loss was entirely due to loss of body fat (BF). Intention to treat analyses demonstrated changes from baseline between rhGH and placebo in BW (-2.16 +/- 4.48 vs. -0.04 +/- 2.67 kg; P = 0.03) and BF (-2.89 +/- 3.76 vs. -0.68 +/- 2.37 kg; P = 0.01). rhGH increased IGF-I from -0.72 to +0.10 SD (P = 0.0001). rhGH increased high-density lipoprotein cholesterol 19% from 1.11 +/- 0.34 to 1.32 +/- 0.28 mmol/liter (P < 0.001). Neither group had changes in fasting glucose, insulin sensitivity, or resting energy expenditure. In conclusion, in obesity, rhGH normalized IGF-I levels, induced loss of BW from BF, and improved lipid profile without untoward effects on insulin sensitivity. PMID- 14764784 TI - Diurnal profiles of testosterone and pituitary hormones suggest different mechanisms for menstrual disturbances in endurance athletes. AB - The aim of this study was to evaluate the diurnal pattern of testosterone and pituitary hormones in endurance female athletes with different types of menstrual disorder. Age- and body mass index-matched groups of endurance athletes with amenorrhea (n = 10) and oligomenorrhea (n = 6), regularly cycling athletes (n = 8), and sedentary controls (n = 8) were compared with respect to 24-h hormonal profiles of testosterone, LH, prolactin (PRL), GH, insulin, IGF binding protein 1 (IGFBP-1), and cortisol. The 24-h hormone profiles in amenorrheic athletes were characterized by decreased LH pulsatility and peak amplitude of PRL and increased baseline levels of GH and cortisol. However, oligomenorrheic athletes displayed a significantly different pattern with higher diurnal testosterone secretion than all other groups. Furthermore, LH, PRL, GH, and cortisol secretions were comparable with regularly menstruating subjects. In the combined group of athletes with menstrual disturbances, diurnal secretions of testosterone, LH, and PRL were positively, whereas cortisol was negatively correlated with the number of menstruations the last year. Although this could be explained by a gradual inhibition of the hypothalamic-pituitary-gonadal axis, our results indicate that the symptoms of amenorrhea and oligomenorrhea may reflect two hormonally distinct conditions. Thus, amenorrheic athletes displayed a hormonal pattern in agreement with hypothalamic inhibition due to energy deficiency, whereas oligomenorrheic athletes demonstrated increased diurnal secretion of testosterone, suggesting a different mechanism, e.g. essential hyperandrogenism. PMID- 14764785 TI - Treatment of hyperthyroidism associated with thyrotropin-secreting pituitary adenomas with iopanoic acid. AB - TSH-secreting tumors comprise less than 2% of all pituitary adenomas. All patients present with hyperthyroidism with a detectable TSH level, and a majority have macroadenomas. Oral cholecystographic agents (e.g. iopanoic acid) potently inhibit the activation of T(4) to the more potent T(3). They have been used successfully to treat primary thyroidal hyperthyroidism and thyroxine overdose. However, they have not been employed in the treatment of central hyperthyroidism. We report, herein, the first two patients with thyrotropinomas, in whom iopanoic acid (Telepaque) has been used perioperatively to safely and rapidly achieve euthyroidism. In case 1, free T(3) index improved from a value of 634 to 175 (normal range 78-162) after 3 d of therapy with iopanoic acid. In case 2, free T(3) by dialysis improved from 697 pg/dl (10.7 pmol/liter) to 195 pg/dl (3.0 pmol/liter) (normal range 210-440 pg/dl; 3.2-6.7 pmol/liter) after 7 d of therapy with iopanoic acid. PMID- 14764786 TI - Serum dehydroepiandrosterone sulfate levels and pubarche in short children born small for gestational age before and during growth hormone treatment. AB - It has been suggested that the programming of the endocrine axes occurs during critical phases of fetal development and will be affected by intrauterine growth retardation. As a result, children born small for gestational age (SGA) might have several hormonal disturbances. In later life, one of the questions that might arise is: Do short children born SGA have higher serum dehydroepiandrosterone sulfate (DHEAS) levels than their peers? Therefore, we compared serum DHEAS levels of 181 short prepubertal children aged 3-9 yr born SGA [birth length (SD score) below -2 for gestational age] with a control group of 170 prepubertal age-matched, normal-statured children born appropriate for gestational age (birth length between -2 and +2 SD score). Because relatively high serum DHEAS levels at a young age might result in a premature pubarche, we investigated the incidence of premature pubarche. We also investigated the association between serum DHEAS levels and bone maturation. In addition, we analyzed whether 1 yr of GH treatment with 1 and 2 mg/m(2).d ( approximately 0.035 and 0.070 mg/kg.d, respectively) had an effect on serum DHEAS levels of prepubertal short SGA children. Serum DHEAS levels of the SGA group were comparable with those of age-matched appropriate for gestational age controls. The incidence of premature pubarche was comparable with that of the normal population. There was a weak negative correlation between serum DHEAS levels and bone maturation after the age of 7 yr. After 1 yr of GH treatment, the increase of serum DHEAS levels was the same for both GH dosage groups and the untreated group. In conclusion, this study shows that small size at birth, which might be a feature of fetal growth restriction, has no effect on serum DHEAS levels before the age of 9 yr. The incidence of premature pubarche is comparable with the normal population. Finally, 1 yr of GH treatment has no effect on serum DHEAS levels. PMID- 14764787 TI - Dose-dependent effects of testosterone on regional adipose tissue distribution in healthy young men. AB - Testosterone supplementation reduces total body adipose tissue (AT), but we do not know whether the effects are uniformly distributed throughout the body or are region specific, or whether they are dose related. We determined the effects of graded doses of testosterone on regional AT distribution in 54 healthy men (18-35 yr) in a 20-wk, randomized, double-blind study of combined treatment with GnRH agonist plus one of five doses (25, 50, 125, 300, or 600 mg/wk) of testosterone enanthate (TE). Total body, appendicular, and trunk AT and lean body mass were measured by dual-energy x-ray absorptiometry, and sc, intermuscular, and intraabdominal AT of the thigh and abdomen were measured by magnetic resonance imaging. Treatment regimens resulted in serum nadir testosterone concentrations ranging from subphysiological to supraphysiological levels. Dose-dependent changes in AT mass were negatively correlated with TE dose at all sites and were equally distributed between the trunk and appendices. The lowest dose was associated with gains in sc, intermuscular, and intraabdominal AT, with the greatest percent increase occurring in the sc stores. At the three highest TE doses, thigh intermuscular AT volume was significantly reduced, with a greater percent loss in intermuscular than sc depots, whereas intraabdominal AT stores remained unchanged. In conclusion, changes in testosterone concentrations in young men are associated with dose-dependent and region-specific changes in AT and lean body mass in the appendices and trunk. Lowering testosterone concentrations below baseline increases sc and deep AT stores in the appendices and abdomen, with a greater percent increase in sc depots. Conversely, elevating testosterone concentrations above baseline induces a greater loss of AT from the smaller, deeper intermuscular stores of the thigh. PMID- 14764788 TI - Trophoblast production of a weakly bioactive human chorionic gonadotropin in trisomy 21-affected pregnancy. AB - Total human chorionic gonadotropin (hCG) is high in maternal serum at 14-18 wk of trisomy 21 (T21)-affected pregnancy, despite low placental hCG synthesis. We sought an explanation for this paradox. We first observed that, in T21-affected pregnancies, maternal serum hCG levels peaked at around 10 wk and then followed the same pattern throughout pregnancy as in controls, albeit at a higher (2.2 fold) level. After delivery, hCG clearance was not significantly different from that in controls. We isolated cytotrophoblasts from 29 T21-affected placentas (12 25 wk) and 13 gestational age-matched control placentas and cultured them for 3 d. In this large series, we confirmed that, in the culture medium of trophoblasts isolated from T21 placentas, hCG secretion was significantly lower (P < 0.003) than in controls, in contrast to the high hCG in maternal serum of the same patients. In T21 cultured trophoblasts, transcripts of sialyltransferase-1 and fucosyltransferase-1 were abnormally high. In corresponding culture medium, hCG was abnormally glycosylated; highly acidic [isoelectric points (pHi) = 4.5] as shown by isoelectric focusing, immunoblotting, and lectin binding; and weakly bioactive (46% of control) as determined using the Leydig cell model. In conclusion, T21 trophoblast cells produced hCG that was weakly bioactive and abnormally glycosylated but whose maternal clearance was unaltered. PMID- 14764789 TI - Estrogen receptor genotypes and their association with the 10-year changes in bone mineral density and osteocalcin concentrations. AB - We conducted a 10-yr prospective study of peak bone mass and its change in 604 women, aged 24-44 yr at study initiation, and related changes in bone mineral density (BMD) and osteocalcin (OCN) concentrations to estrogen receptor (ER) alpha gene polymorphisms in 442 of these women. We examined the association of ER alpha PvuII and XbaI polymorphisms with the 10-yr change in lumbar spine (LS) and femoral neck (FN) BMD, measured by densitometry, as well as serum OCN levels, after accounting for weight and menstrual status change. The women were members of the Michigan Bone Health Study, a population-based longitudinal study of BMD. There was a linear loss of LS BMD and curvilinear loss of FN BMD from peak bone mass over a 10-yr period. Women homozygous for the ER alpha gene variant without an XbaI restriction site (XbaI -/- genotype) had higher FN BMD and less change in LS over time. Women homozygous for the ER alpha gene variant without a PvuII restriction site (PvuII -/- genotype) had less LS BMD change over time as well as higher FN BMD. However, this higher FN BMD was dependent upon the rate of bone turnover as estimated from serum OCN change over time. The ER alpha genotype associations were statistically significant in explaining the rate of perimenopausal bone loss and its turnover; however, BMI or becoming postmenopausal contributed more to the magnitude of the difference in bone change. PMID- 14764790 TI - The effects of atorvastatin on endothelial function in diabetic patients and subjects at risk for type 2 diabetes. AB - We have investigated the effect of atorvastatin on the endothelial function of patients with diabetes and subjects at risk for type 2 diabetes in a 12-wk, prospective, randomized, placebo-controlled, double-blind clinical trial. The flow- mediated dilation (FMD; endothelium dependent) and nitroglycerin-induced dilation (endothelium independent) in the brachial artery and the vascular reactivity at the forearm skin were measured. FMD improved in the atorvastatin treated, at-risk subjects [median (25-75 percentile), 7.2% (2.9-9.6%) at exit visit vs. 6.6% (2.9-9.5%) at baseline; P < 0.05]. A similar improvement of FMD was found in atorvastatin-treated diabetic patients [median (25-75 percentile), 5.6 (3.9-7.9) at exit visit vs. 4.2 (3.2-7.2) at baseline; P = 0.07]. No changes were observed in nitroglycerin-induced dilation and the microcirculation reactivity measurements in either group. In the at-risk group, there was a decrease in the C-reactive protein [median (25-75 percentile), 0.12 mg/dl (0.07 0.27 mg/dl) at exit visit vs. 0.24 mg/dl (0.07-0.35 mg/dl) at baseline; P < 0.05] and TNF alpha [median (25-75 percentile), 2.6 pg/ml (1.8-4.1 pg/ml) at exit visit vs. 4.4 pg/ml (3.6-6.0 pg/ml) at baseline; P < 0.05] in the atorvastatin-treated patients, whereas in the diabetes group, a decrease in endothelin-1 (mean +/- SD, 0.97 +/- 0.29 pg/ml at exit visit vs. 1.19 +/- 0.42 pg/ml at baseline; P < 0.05) and plasminogen activator inhibitor-1 [median (25-75 percentile), 18 ng/ml (9-24 ng/ml) at exit visit vs. 27 ng/ml (7-41 ng/ml) at baseline; P < 0.05] were observed. We conclude that atorvastatin improves endothelial function and decreases levels of markers of endothelial activation and inflammation. PMID- 14764791 TI - Common variants in glutamine:fructose-6-phosphate amidotransferase 2 (GFPT2) gene are associated with type 2 diabetes, diabetic nephropathy, and increased GFPT2 mRNA levels. AB - Increased flux of glucose through the hexosamine biosynthetic pathway has been implicated in insulin resistance, altered insulin secretion, and diabetic nephropathy. Glutamine:fructose-6-phosphate amidotransferase (GFPT), the rate limiting enzyme in hexosamine biosynthesis, is encoded by the unlinked but highly homologous genes GFPT1 and GFPT2. We tested the hypothesis that GFPT2 sequence variation contributed to the susceptibility to type 2 diabetes mellitus (T2DM) and diabetic nephropathy in Caucasian and African-American individuals. We identified 11 single nucleotide polymorphisms (SNPs), of which seven were common. A single variant in exon 14, I471V, altered the amino acid sequence, is conserved between human and mouse genes, and was associated with T2DM among Caucasians (P = 0.05). A trend to an association was noted with diabetic nephropathy among African-American individuals (P = 0.15). Several variants in the 3' untranslated region (UTR) and exon 18 were also associated with T2DM in Caucasian individuals (P < 0.05), and the SNP in the 3' UTR was associated with diabetic nephropathy in African-American subjects (P = 0.047). GFPT2 mRNA levels in transformed lymphocytes from study subjects were significantly increased among African American subjects compared with Caucasian individuals, regardless of diagnosis. Furthermore, the associated allele of the 3' UTR SNP was approximately 2-fold overexpressed. We propose that the 3' UTR variant results in increased GFPT2 mRNA levels with resultant increased hexosamine flux. The I471V variant may contribute to altered protein function or may simply be in linkage disequilibrium with the 3' UTR. PMID- 14764792 TI - Adiponectin gene expression and plasma values in obese women during very-low calorie diet. Relationship with cardiovascular risk factors and insulin resistance. AB - Adiponectin, a newly discovered adipose-tissue-specific protein, is thought to be involved in the regulation of insulin action. The aim of the present study was to determine whether adiponectin contributes to the improvement in insulin sensitivity during very-low-calorie diet (VLCD). Biopsies of sc abdominal adipose tissue and blood sampling for analysis of plasma adiponectin and related hormones and metabolites were performed before and at the end of a 4-wk VLCD in 33 nonmorbidly obese women (body mass index, 34.4 +/- 4.1 kg/m(2)). VLCD produced a decrease in weight (7.1 +/- 0.4 kg) and in insulin and leptin levels and led to an improvement in insulin sensitivity. Adiponectin gene expression and plasma levels were not modified during calorie restriction. Before VLCD, we found negative correlations between plasma adiponectin and variables related to the metabolic syndrome. Adiponectin mRNA levels showed a negative correlation with lipoprotein a plasma values. The correlations observed before VLCD were not found after VLCD. The data suggest that adiponectin is related to the protection against the metabolic syndrome but is not involved in the regulation of VLCD induced improvement of insulin sensitivity. PMID- 14764793 TI - Is the decreased high-density lipoprotein cholesterol in the metabolic syndrome due to cellular lipid efflux defect? AB - The metabolic syndrome (MS) is associated with cardiovascular disease. The low high-density lipoprotein cholesterol (HDL-C) seen in the MS is associated with increased hepatic secretion of apolipoprotein B-containing lipoproteins. Patients with low HDL-C and abnormal cellular lipid efflux due to ABCA1 gene defects (Tangier disease) also have elevated plasma triglycerides. In the present study, we examined the cellular cholesterol and phospholipid efflux in patients with low HDL-C and features of the MS. Forty-four patients with a HDL-C below the fifth percentile for age and gender were selected. The MS was defined by a low HDL-C and at least two additional features: body mass index at least 30 kg/m(2), plasma triglycerides at least 150 mg/dl, fasting glucose at least 110 mg/dl, and blood pressure at least 130/85 mm Hg. Cellular lipid efflux was examined on fibroblasts obtained from study subjects, nine normal controls and six subjects with Tangier disease. In 22 patients identified with the MS, HDL-C was 21 +/- 7 mg/dl, triglyceride levels were 340 +/- 157 mg/dl, and cellular cholesterol and phospholipid efflux were 107 +/- 18% and 105 +/- 17% of controls, respectively. No patient with the MS and low HDL-C showed a cellular lipid efflux defect. We conclude that primary cellular lipid efflux defects do not contribute to the low HDL-C frequently encountered in the MS. PMID- 14764794 TI - Hypoadiponectinemia is associated with impaired endothelium-dependent vasodilation. AB - Adiponectin may have an antiatherogenic effect by reducing endothelial activation. We hypothesized that plasma adiponectin levels were correlated with endothelial function. Plasma adiponectin level was determined by an in-house RIA assay using a rabbit polyclonal antibody in 73 type 2 diabetic patients and 73 controls. Endothelium-dependent and independent vasodilation of the brachial artery was measured by high-resolution vascular ultrasound. Plasma adiponectin level was lower in diabetic patients than in controls (4.73 +/- 1.96 vs. 7.69 +/- 2.80 microg/ml, respectively; P < 0.001), and they also had impaired endothelium dependent (5.6 +/- 3.6 vs. 8.6 +/- 4.5%, respectively; P < 0.001) and independent vasodilation (13.3 +/- 4.9 vs. 16.5 +/- 5.6%, respectively; P < 0.001). Plasma adiponectin correlated with endothelium-dependent vasodilation in controls (P = 0.02) and diabetic patients (P = 0.04). On general linear-model univariate analysis, brachial artery diameter, the presence of diabetes, plasma adiponectin, and high-density lipoprotein were significant independent determinants of endothelium-dependent vasodilation. In vitro experiments showed that endothelial cells expressed adiponectin receptors, and adiponectin increased nitric oxide production in human aortic endothelial cells. In conclusion, low plasma adiponectin level is associated with impaired endothelium-dependent vasodilation, and the association is independent of diabetes mellitus. Adiponectin may act as a link between adipose tissue and the vasculature. PMID- 14764795 TI - First trimester placental growth factor and soluble fms-like tyrosine kinase 1 and risk for preeclampsia. AB - An imbalance of pro- and antiangiogenic factors may lead to preeclampsia (PE). In this prospective nested case-control study, we investigated whether first trimester serum levels of placental growth factor (PlGF), a potent angiogenic factor, and its soluble inhibitor, soluble fms-like tyrosine kinase 1 (sFlt1), distinguished women who developed PE (n = 40) from those who developed gestational hypertension (n = 40), delivered a small for gestational age (SGA) newborn (n = 40), or completed a full term normal pregnancy (n = 80). Compared with controls, serum PlGF levels were lower among women who developed PE (23 +/- 24 pg/ml vs. 63 +/- 145 pg/ml; P < 0.01) or gestational hypertension (27 +/- 19 pg/ml; P = 0.03), or who delivered a SGA newborn (21 +/- 16 pg/ml; P < 0.01). In contrast, serum sFlt1 levels did not markedly differ between the groups: PE, 1048 +/- 657 pg/ml; gestational hypertension, 942 +/- 437 pg/ml; SGA newborns, 1011 +/ 479 pg/ml; and normal controls, 973 +/- 490 pg/ml. Multivariable analysis adjusting for potential confounders and serum sFlt1 levels demonstrated a 3.7 fold (95% confidence interval, 1.2-12.5) increase in risk for PE for every log unit decrease in serum levels of PlGF compared with controls. Analyses for gestational hypertension and SGA were not significant. Examined in tertiles, the risk for PE was increased 28.7-fold (95% confidence interval, 2.3-351.0) in the third (<12 pg/ml) compared with the first (>39 pg/ml) PlGF tertile. First trimester serum levels of PlGF and sFlt1 may identify women at high risk for PE. PMID- 14764796 TI - Plasma lipid and lipoprotein levels during the follicular and luteal phases of the menstrual cycle. AB - Estrogen levels are higher during the luteal compared with the follicular phase of the menstrual cycle. It was hypothesized that the luteal compared with the follicular phase has a lipid and lipoprotein profile associated with decreased coronary heart disease (CHD) risk. This was tested using well-defined data from healthy, well-characterized premenopausal Caucasian women under very controlled metabolic conditions. The percent differences in lipid, lipoprotein, and sex hormone levels between the follicular and luteal phases were estimated using generalized estimating equations after adjusting for age, body mass index, calendar time, and season. The low-density lipoprotein cholesterol (LDL-C) level was 6.2% lower (P = 0.015), and the total cholesterol/high-density lipoprotein cholesterol (HDL-C) and LDL-C/HDL-C ratios were 5.1% (P = 0.0006) and 8.4% (P = 0.002) lower, respectively, during the luteal phase. Levels of estradiol and other estrogens were significantly higher (by>100% each; P < 0.0001 in all cases) in the luteal phase. These findings support the study hypothesis. Fluctuations in levels of LDL-C and the total cholesterol/HDL-C and LDL-C/HDL-C ratios between menstrual cycle phases need to be considered in the screening and medical monitoring of premenopausal women, especially those with borderline levels. Although small, such fluctuations may prove to be clinically significant in the long run. Studies involving premenopausal women need to more clearly define and validate menstrual cycle phase in the design and interpretation of study results. PMID- 14764797 TI - The hormonal phenotype of Nonclassic 3 beta-hydroxysteroid dehydrogenase (HSD3B) deficiency in hyperandrogenic females is associated with insulin-resistant polycystic ovary syndrome and is not a variant of inherited HSD3B2 deficiency. AB - To test our hypothesis that the hormonal phenotype of mild 3beta-hydroxysteroid dehydrogenase (HSD3B) deficiency in hyperandrogenic females (HF) is related to insulin-resistant polycystic ovary syndrome (PCOS), we compared insulin sensitivity and gonadotropin secretion in HF with compromised ( downward arrow ) adrenal HSD3B phenotype despite normal HSD3B2 genes (n = 6) to those in HF with classic PCOS (n = 9) of similar ages (14-36 yr). The same was examined in premature pubarche (PP) girls with (n = 4) and without the descending HSD3B phenotype (n = 5). The descending HSD3B phenotype was defined by ACTH-stimulated Delta(5)-precursor steroid levels and Delta(5)-precursors to Delta(4)-product steroid ratios higher than those in normal females (n = 30 for adult, n = 12 for pubertal). Classic PCOS HF had elevated testosterone levels and normal ACTH stimulated hormonal profiles. The insulin sensitivity index determined by the frequently sampled iv glucose-tolbutamide test (FSIVGTT) in all HF with descending HSD3B phenotype and in all HF with classic PCOS, regardless of body mass index (BMI), was lower than in all eight normal BMI and five high BMI normal females. Integrated incremental insulin determined by FSIVGTT, the area under the curve for insulin, and fasting and 2 h glucose load insulin levels determined by an oral glucose tolerance test in both HF groups were higher (P < 0.01-0.0001) than those in normal females with normal or high BMI. LHRH-stimulated LH levels and LH/FSH ratios in both HF groups were higher (P < 0.01) than those in normal females. No statistical differences were found in the insulin sensitivity and gonadotropin parameter between the two PP girl groups. The insulin sensitivity index in each half of PP girls with the descending HSD3B phenotype was lower than or similar to that in control PP girls with a similar weight length index. The fasting glucose to insulin ratio in three of four PP girls with the descending HSD3B phenotype was lower than that in control PP girls, but one of four with the descending HSD3B phenotype had a higher fasting glucose to insulin ratio than the control PP girls. The findings of insulin sensitivity and gonadotropin data in both HF with the descending HSD3B phenotype and classic PCOS indicate significant insulin resistance and LH hypersecretion in both. These suggest that the descending HSD3B phenotype in HF is associated with a variant of insulin resistant PCOS. The variable insulin sensitivity parameter in the small number of PP girls with the descending HSD3B phenotype warrants a further large scale study to examine this phenotype association with childhood insulin resistance. PMID- 14764798 TI - Induction of beta-cell rest by a Kir6.2/SUR1-selective K(ATP)-channel opener preserves beta-cell insulin stores and insulin secretion in human islets cultured at high (11 mM) glucose. AB - In health, most insulin is secreted in pulses. Type 2 diabetes mellitus (TTDM) is characterized by impaired pulsatile insulin secretion with a defect in insulin pulse mass. It has been suggested that this defect is partly due to chronic overstimulation of beta-cells imposed by insulin resistance and hyperglycemia, which results in depletion of pancreatic insulin stores. It has been reported that in TTDM overnight inhibition of insulin secretion (induction of beta-cell rest) leads to quantitative normalization of pulsatile insulin secretion upon subsequent stimulation. Recently, decreased orderliness of insulin secretion has been recognized as another attribute of impaired insulin secretion in TTDM. In the current studies we sought to address at the level of the isolated islet whether chronic elevated glucose concentrations induce both defects involved in impaired insulin secretion in TTDM: deficiency and decreased orderliness of insulin secretion. We use the concept of beta-cell rest, induced by a novel beta cell selective K(ATP)-channel opener (KCO), NN414 (6-chloro-3-(1 methylcyclopropyl)amino-4H-thieno[3,2-e]-1,2,4-thiadiazine 1,1-dioxide), to test whether preservation of insulin stores leads to normalization of both processes in response to glucose stimulation. Human islets were isolated from three cadaveric organ donors and studied in perifusion experiments and static incubation. Acute activation of K(ATP)-channels suppressed insulin secretion from perifused human islets by approximately 90% (P < 0.0001). This KCO also inhibited glucagon secretion in a dose-dependent manner (P = 0.01). Static incubation at 11 and 16 vs. 4 mM glucose for 96 h decreased islet insulin stores by approximately 80% and 85% (P < 0.0001, respectively). In subsequent perifusion experiments, total insulin secretion ( approximately 30%; P < 0.01) from these islets and insulin pulse mass ( approximately 40%; P < 0.05) were both decreased (11 vs. 4 mM). The inhibition of insulin secretion during static incubation with KCO reduced the loss of islet insulin stores in a dose-dependent manner (P < 0.0001) and resulted in increased total insulin secretion (2.6-fold; P < 0.01) and insulin pulse mass (2.5-fold; P < 0.05) during subsequent perifusion. The orderliness of insulin secretion was significantly reduced after chronic incubation of human islets at 11 mM glucose (P = 0.04), but induction of beta cell rest at 11 mM failed to normalize the regularity of insulin secretion during subsequent perifusion. We conclude that physiological increased glucose concentrations (11 mM), which are frequently observed in diabetes, lead to a loss of islet insulin stores and defective pulsatile insulin secretion as well as reduced orderliness of insulin secretion. Induction of beta-cell rest by selective activation of beta-cell K(ATP)-channels preserves insulin stores and pulsatile insulin secretion without restoring the orderliness of insulin secretion. Therefore, the concept of beta-cell rest may provide a strategy to protect beta-cells from chronic overstimulation and to improve islet function. Impaired glucose-regulated insulin secretion in TTDM may, however, partially involve mechanisms that are distinct from insulin stores and insulin secretion rates. PMID- 14764799 TI - Serum interleukin-18 concentrations are increased in the polycystic ovary syndrome: relationship to insulin resistance and to obesity. AB - Low-grade chronic inflammation is involved in the pathogenesis of the metabolic syndrome and atherosclerosis, and serum levels of inflammatory cytokines are useful cardiovascular risk markers. We have studied serum IL-18 concentrations in women with polycystic ovary syndrome (PCOS), focusing on its relationship with obesity and indexes of insulin resistance. Sixty consecutive women with PCOS and 34 healthy women were recruited. Serum levels of IL-18 and lipid and hormone profiles were measured. The insulin sensitivity index was calculated from glucose and insulin concentrations during an oral glucose tolerance test. Data were submitted to a multivariate general linear model introducing age as a covariate. Serum IL-18 levels were increased in PCOS patients compared with controls (P = 0.031) and in obese women compared with lean women (P = 0.018). No interaction between PCOS and obesity was found, suggesting that the influence of PCOS on serum IL-18 concentrations studied here was not different in lean women compared with obese women and that the influence of obesity on serum IL-18 concentrations was the same in the PCOS and control groups. Serum IL-18 levels correlated, after logarithmic transformation, with body mass index (r = 0.38; P < 0.0002), waist-to hip ratio (r = 0.33; P < 0.001), and total testosterone levels (r = 0.24; P < 0.02), and inversely with the insulin sensitivity index (r = -0.23; P < 0.03). In conclusion, PCOS and obesity induce an increase in serum IL-18 levels, which are also associated with several indexes of global and visceral adiposity and with insulin resistance. PMID- 14764800 TI - Tumor necrosis factor-alpha inhibits trophoblast migration through elevation of plasminogen activator inhibitor-1 in first-trimester villous explant cultures. AB - We have tested the hypothesis that elevated concentrations of TNF alpha could impair trophoblast invasion. Using first-trimester placental explant cultures, we have demonstrated that the cytokine inhibits in vitro migration of extravillous trophoblasts (EVT) on collagen I, and invasion through Matrigel. To elucidate the underlying mechanism, proliferation and differentiation of EVT in vitro were analyzed by immunohistochemistry of serial sections, Western blotting, zymography, ELISA, and RT-PCR from RNA pools of mechanically separated cell populations. At 24 h of cultivation in the presence or absence of TNF alpha, anchorage and proliferation of trophoblasts had occurred to generate cell columns containing viable, post-mitotic, differentiated EVT [positive for integrins alpha 1 and alpha 5, matrix metalloproteinase (MMP)-2, and human leukocyte antigen-G1; negative for proliferating cellular nuclear antigen, cytokeratin 18 neoepitope, and in 5-Bromo-2-deoxy-uridine labeling]. At 72 h, control cells had broken away from the column to migrate through the extracellular matrix; whereas, in contrast, TNF alpha-treated EVT remained as contiguous cell columns, despite increased MMP-9 expression. Thus, in vitro MMP9 activity appears not to be essential for trophoblast migration. Expression of plasminogen activator inhibitor (PAI)-1 was elevated in TNF alpha-treated EVT, and adding antibodies that inhibit PAI-1 activity restored migration, whereas tissue-inhibitor-of metalloproteinases-1-blocking antibodies were ineffective. Induction of PAI-1 by TNF alpha could be related to restricted trophoblast invasion in preeclampsia. PMID- 14764801 TI - Hepatocyte growth factor/Met system promotes endometrial and endometriotic stromal cell invasion via autocrine and paracrine pathways. AB - Endometrial stromal cells reportedly have a role in the initial invasion of endometrial tissue into the peritoneum. Hepatocyte growth factor (HGF), which is a ligand for the c-met protooncogene product (Met), stimulates proliferation and invasion of a large number of cells. In this study we investigated the role of the HGF/Met system in the pathogenesis of endometriosis. HGF concentrations in the peritoneal fluid of patients with endometriosis were significantly higher than in those without endometriosis and correlated positively with revised American Society of Reproductive Medicine scores. We showed that the peritoneum and endometriotic stromal cells may be major sources of HGF in peritoneal fluid. Endometrial and endometriotic stromal cells expressed the Met receptor, which was activated by endogenous and exogenous HGF. HGF enhanced stromal cell proliferation and invasion. We also demonstrated that the HGF-stimulated stromal cell invasion was due in part to the induction of urokinase-type plasminogen activator, a member of the extracellular proteolysis system. In conclusion, the HGF/Met system is involved in the pathogenesis of endometriosis by promoting stromal cell proliferation and invasion of shed endometria and endometrial lesions via autocrine and paracrine pathways. PMID- 14764802 TI - Reduced serum glycodelin and insulin-like growth factor-binding protein-1 in women with polycystic ovary syndrome during first trimester of pregnancy. AB - The polycystic ovary syndrome (PCOS) is associated with an increased rate of early pregnancy loss (EPL). Hyperinsulinemia is an independent risk factor for EPL and has been found to decrease levels of glycodelin and IGF binding protein-1 (IGFBP-1), two major endometrial proteins. We hypothesized that serum glycodelin IGFBP-1 concentrations would be reduced in women with PCOS during the first trimester of pregnancy. Fasting serum insulin, glycodelin, and IGFBP-1 were measured, and oral glucose tolerance tests were performed in 72 women with PCOS and 62 normal women. Each woman was seen once and assigned to one of three gestational groups: wk 3-5, 6-8, and 9-11. The insulin sensitivity index during oral glucose tolerance test was lower in women with PCOS compared with normal women throughout the first trimester (P < 0.0001). Both serum glycodelin and IGFBP-1 were markedly lower in women with PCOS (for glycodelin: wk 3-5, P < 0.0001; wk 6-8, P = 0.03; wk 9-11, P = 0.19; and for IGFBP-1: wk 3-5 and 6-8, P < 0.0001; wk 9-11, P = 0.0003). Comparing women with PCOS who experienced EPL with those who did not, serum glycodelin was significantly lower during wk 3-8 (P < 0.02) and serum IGFBP-1 during wk 9-11 (P = 0.003). During the first trimester, serum glycodelin and IGFBP-1 concentrations are markedly decreased in PCOS, implicating endometrial epithelial and stromal dysfunction during periimplantation and early pregnancy as a possible mechanism for EPL in PCOS. These decreases are likely to be secondary to hyperinsulinemia and reduced insulin sensitivity. PMID- 14764803 TI - Contrasting negative-feedback control of endogenously driven and exercise stimulated pulsatile growth hormone secretion in women and men. AB - GH represses its own secretion via rapid and reversible feedback exerted at key hypothalamic loci. The primary mechanisms include stimulation of somatostatin release and inhibition of GHRH outflow. Autoinhibition is prominent in the adult male rat but diminutive in the female animal. The sex contrast reflects important differences in central neuropeptide signaling in this species. No comparable insights into gender-specific control of GH autofeedback are available in the human. To examine this issue, we quantitated acute recombinant human (rh)GH induced inhibition of baseline (resting) and aerobic exercise-stimulated GH secretion in healthy young men (n = 8) and early follicular-phase women (n = 6). Each subject underwent four fasting, morning inpatient infusion studies in a prospectively randomized, placebo-controlled, double-blind, within-subject cross over design. The feedback paradigm comprised 6-min bolus iv infusion of saline or rhGH (10 microg/kg) followed in 120 min by rest or submaximal aerobic (individually calibrated) bicycle ergometry for 30 min. Concomitantly, blood was sampled every 10 min for 6 h, and sera were submitted to immunochemiluminometric GH assay (sensitivity 0.005 microg/liter). Biexponential deconvolution analysis was applied to estimate stimulated GH secretory-burst mass (microg/liter per 90 min after onset of exercise or rest). Women and men had statistically comparable serum estradiol but unequal testosterone concentrations. Repeated-measures ANOVA documented a significant three-way interaction among gender, stimulus type (rest or exercise), and feedback status (saline or rhGH injection) in determining GH secretory-burst mass (P = 0.008). There were prominent two-factor interactions among gender and exercise (P < 0.001); gender and rhGH-induced negative feedback (P = 0.002); and exercise and rhGH feedback (P = 0.006). Gender comparisons disclosed that women, compared with men, maintain 20-fold higher GH secretory burst mass at rest (P < 0.001); 40-fold less stimulation of pulsatile GH release by exercise than rest (P < 0.001); and 20-fold greater inhibition of GH secretory burst mass by rhGH than saline at rest (P < 0.05). Observed feedback contrasts by sex were specific, inasmuch as gender did not affect absolute estimates of exercise-stimulated GH secretion (microg/liter/90 min); nadir GH concentrations (microg/liter) enforced by rhGH infusion; and the time latency (min) to manifest maximal inhibition after rhGH injection. In summary, the present clinical investigation unmasks: 1) markedly greater fractional feedback inhibition of pulsatile GH secretion by rhGH in young women than men; and 2) partial resistance of the aerobic-exercise stimulus to GH autofeedback in both women and men. We postulate that sex-steroid-specific control of somatostatin and GHRH outflow may mediate the former gender contrasts, whereas unknown (gender-independent) factors may determine the capability of exercise to significantly antagonize GH autoinhibition. PMID- 14764804 TI - Insulin-degrading activity in wound fluid. AB - Patients with diabetes are at great risk of developing lower extremity ulcers. The management of diabetic foot ulcers typically includes early recognition and appropriate clinical care. Recent advances in wound treatment include topical growth factor therapy, which has been successful in diabetic wounds. Growth factors are decreased in wound fluid; this may be due to decreased supply, increased binding, or increased degradation of the naturally occurring growth factors. This study investigates the activity of the insulin-degrading enzyme in wound fluid. Wound fluid was obtained from patients with (n = 17) and without (n = 4) diabetes. Insulin degradation was assayed by incubating [(125)I]insulin with wound fluid and precipitation in trichloroacetic acid. Fluid from nondiabetics degraded 2.22 +/- 0.73%, whereas diabetic fluid degraded significantly more (6.13 +/- 1.48%; P < 0.05). In patients with diabetes, the degradation of insulin by wound fluid correlated with glucose control (hemoglobin A(1c); r(2) = 0.5353; P < 0.001), and patients with worse outcomes (i.e. amputation) had higher wound fluid insulin degradation. The biochemical characteristics of insulin degradation in the wound fluid were consistent with the characteristics of insulin-degrading enzyme. These data suggest that glucose control is a critical factor in wound healing, but a reduction in the insulin-degrading activity in the wound fluid is also a potential therapeutic target. PMID- 14764805 TI - Tibolone and its metabolites induce antimitogenesis in human coronary artery smooth muscle cells: role of estrogen, progesterone, and androgen receptors. AB - Tibolone, a hormone replacement drug, protects postmenopausal women against osteoporosis and climacteric symptoms without inducing adverse effects on the endometrium and breast. Compared with other estrogens, little is known about the cardiovascular effects of tibolone. Because abnormal growth of smooth muscle cells (SMCs) is a prerequisite for coronary artery disease, here we investigated the effects of tibolone on SMC growth. We examined the effects of tibolone and its metabolites on human arterial SMC growth (DNA synthesis, cellular proliferation, cell migration, collagen synthesis) and MAPK expression. Fetal calf serum-induced SMC growth, phosphorylated MAPK expression, and platelet derived growth factor-induced SMC-migration were concentration-dependently inhibited by tibolone and its endogenous estrogenic and progestogenic/androgenic metabolites in the following order of potency: Delta 4-tibolone>3 beta-OH tibolone congruent with 3 alpha-OH-tibolone. The antimitogenic effects of tibolone were partially blocked by ER antagonist (ICI182780), progesterone receptor antagonist (RU486) but not by the androgen receptor antagonist (flutamide); moreover, RU486 was more potent than ICI182780. The antimitogenic effects of tibolone were completely blocked by RU486 plus ICI182780. In addition, the inhibitory effects of equimolar concentrations of the three tibolone metabolites summed up to the inhibitory effects of tibolone. In conclusion, tibolone inhibits SMC growth and MAPK phosphorylation via both its estrogenic and progestogenic metabolites, and these inhibitory effects involve both progesterone and ERs. Hence, tibolone may induce antivasoocclusive actions and protect women against coronary artery disease. PMID- 14764806 TI - Administration of dihydrotestosterone to rhesus monkeys inhibits gonadotropin stimulated ovarian steroidogenesis. AB - Androgens, in addition to serving as a substrate for estrogen biosynthesis, exert autocrine/paracrine actions on ovarian function. However, much of the information regarding the actions of androgens on the ovary has been obtained using rodents, and the extent to which these results can be extrapolated to higher primates is uncertain. The current study was initiated to determine the effects of dihydrotestosterone (DHT) and testosterone (T) on the responsiveness of the rhesus monkey ovary to exogenous FSH and LH in vivo. Rhesus monkeys whose spontaneous gonadotropin secretion was interrupted with a GnRH antagonist received s. . implants of either DHT or T for 5 d before and continuing throughout a 15-d i.v. infusion of human FSH and LH. Neither T nor DHT treatment synergized with FSH/LH to stimulate estrogen production or increases in ovarian weight. Rather, administration of DHT significantly reduced estrogen secretion and the augmentation of ovarian weight in response to exogenously administered FSH and LH. These results indicate that high concentrations of DHT are antagonistic to gonadotropin-stimulated ovarian function in primates. PMID- 14764807 TI - Nocturnal secretory dynamics of inhibin B and testosterone in pre- and peripubertal boys. AB - To investigate the secretory dynamics of testosterone and inhibin B, we collected samples every 20 min from 2000 h to 0800 h in 20 boys. Boys in group 1 (n = 5) were aged less than 8 yr, group 2 (n = 5) were aged more than 8 yr but 1.5 yr or more before pubertal onset, group 3 (n = 5) were studied 1.0 yr or less before pubertal onset, and group 4 (n = 5) were in early puberty. Testosterone increased after midnight in peripubertal boys, coinciding with the onset of LH pulsatility, and showed a pulsatile pattern in 6 of 10 of these boys. Cross-correlation analysis indicated significant temporal coupling between LH and testosterone. Inhibin B was higher in groups 3 and 4, compared with groups 1 and 2 (P < 0.01) and showed a downward trend overnight with no evidence of pulsatility and no evidence of short-term interactions with LH, FSH, or testosterone. Inhibin B and LH nocturnal means were both inversely correlated with time before pubertal onset (r(s) > or = -0.85, P < 0.01). Only LH nocturnal mean and amplitude, respectively, contributed independently to prediction of testosterone and inhibin B nocturnal means, explaining 71 and 65% of their variability. We conclude that both testosterone and inhibin B are related to nocturnal LH release in peripubertal boys but over different time scales. PMID- 14764808 TI - A follow-up linkage study for quantitative trait loci contributing to obesity related phenotypes. AB - We have recently reported a whole-genome scan in a sample of 630 subjects from 53 extended pedigrees, in which several genomic regions that may contain quantitative trait loci (QTLs) for obesity were suggested. In the present study, with an attempt to confirm our previous findings, we performed a follow-up linkage study in an expanded sample of 79 pedigrees with 1816 subjects (including expanded previous 53 pedigrees and 26 newly recruited pedigrees containing 1058 subjects). A new set of microsatellite markers spanning previously identified regions were selected, with the average genomic distance narrowed from approximately 10 cM to approximately 5 cM in this study. Using a variance component method, we performed two- and multipoint linkage analyses in the following three sample sets: expanded previous 53 pedigrees (758 subjects), 26 new pedigrees, and 79 total pedigrees. For body mass index, analyses of the expanded 53 pedigrees attained a LOD score of 2.32 near marker D1S468 in two point analysis and a maximum LOD score (MLS) of 2.21 in multipoint analysis; 2q14 near marker D2S347 attained a LOD score of 3.42 in two-point analysis and a MLS of 3.93 in multipoint analysis. The linkage peaks at 1p36 and 2q14 were further supported in the analyses of all 79 pedigrees, with multipoint MLS being 1.38 and 0.90, respectively. For fat mass, genomic region 6q27 achieved a LOD score of 1.24 in two-point analysis and an MLS of 0.92 in multipoint analysis in all 79 pedigrees. Our data support that 1p36, 2q14, and 6q27 are promising regions that may harbor QTLs for obesity phenotypes. PMID- 14764809 TI - Estrogen enhances cystatin C expression in the macaque vagina. AB - Cystatin C is a secreted inhibitor of cysteine proteinases that participates in extracellular matrix remodeling. Whether hormones affect its expression in the vagina was unknown. Consequently, we examined the effects of estradiol (E(2)), progesterone (P), and raloxifene on vaginal cystatin C in rhesus macaques. In experiment 1, ovariectomized animals were treated sequentially with E(2) (14 d) and E(2) + P (14 d) to induce 28-d menstrual cycles. Vaginal samples were collected on d 6, 8, 14, and 28 of the induced cycle. Some cycled animals were deprived of both E(2) + P for 28 d. In experiment 2, ovariectomized animals were treated for 5 months with E(2) alone, E(2) + P, raloxifene, or left untreated. Total RNA from the vaginal wall was analyzed for the cystatin C transcript with a commercially prepared cDNA array and semiquantitative RT-PCR. Vaginal cryosections were analyzed by in situ hybridization for cystatin C transcript and by immunocytochemistry for the protein. E(2) treatment significantly (5-fold; P < 0.05) increased expression of cystatin C transcript over the levels in the hormone-deprived controls, and cotreatment with P (E(2) + P) blocked this effect. Raloxifene treatment did not affect cystatin C expression. In situ hybridization and immunocytochemistry revealed that cystatin C was localized in fibroblasts and smooth muscle cells throughout the vaginal wall but not in smooth muscle cells of arteries or levator ani myocytes. In summary, E(2) increased vaginal cystatin C expression in the fibroblasts and smooth muscle bundles, P suppressed this effect, and raloxifene had no effects on cystatin C. Elevated cystatin C, by suppressing cysteine proteinase activity, may strengthen the vaginal wall and mitigate the potential for pelvic floor prolapse. PMID- 14764810 TI - Glucocorticoid sensitivity is determined by a specific glucocorticoid receptor haplotype. AB - Differences in glucocorticoid (GC) sensitivity may underlie both common diseases (e.g. hypertension) and variability in response to treatment with GCs (e.g. asthma). We tested the potential involvement of the GC receptor (GR) gene in mediating GC sensitivity using haplotype analysis and a low-dose dexamethasone suppression test. Linkage disequilibrium across the GR gene was determined in 216 U.K. Caucasians, and 116 had a 0.25-mg overnight dexamethasone suppression test. Very strong linkage disequilibrium was observed across the GR gene with only four haplotypes accounting for 95% of those observed. Haplotype pattern mining and linear regression analyses independently identified a three-marker haplotype, across intron B, to be significantly associated with low postdexamethasone cortisol (P = 0.03). Carriage of this haplotype occurred in 41% of the individuals with low postdexamethasone cortisol vs. 23% in the combined other quartiles (odds ratio 2.4, 95% confidence interval 0.9-6.3, P = 0.05). This is the first comprehensive, haplotype based analysis of the GR gene. A three-point haplotype, within intron B, is associated with enhanced sensitivity to GCs. This haplotype may help predetermine variation in clinical response to GC therapy and also assist the understanding of diseases related to GC production. PMID- 14764811 TI - Promoter polymorphism in PCK1 (phosphoenolpyruvate carboxykinase gene) associated with type 2 diabetes mellitus. AB - We sequenced the promoter and coding regions of PCK1 encoding cytosolic phosphoenolpyruvate carboxykinase from genomic DNA of subjects with type 2 diabetes mellitus (DM). We found nine single nucleotide polymorphisms (SNPs) that were present with varying allele frequencies and pairwise linkage disequilibrium relationships in different ethnic groups. The -232C-->G promoter SNP was within a cis-acting element required for basal and cAMP-mediated PCK1 gene transcription. The expression of a luciferase reporter construct containing -232G in three different cell lines showed significantly increased basal expression with no down regulation by insulin compared with a construct containing -232C. The odds ratios for type 2 DM among subjects with one or two copies of -232G compared with 232C/C homozygotes were 1.9 (95% confidence interval, 1.2-3.0) in a Canadian aboriginal sample and 2.8 (95% confidence interval, 1.7-4.7) in a Caucasian sample. Thus, we report a promoter SNP in PCK1 that was resistant to down regulation by insulin in vitro and was associated with type 2 DM in two independent study samples. PMID- 14764812 TI - Genetic screening for melanocortin-4 receptor mutations in a cohort of Italian obese patients: description and functional characterization of a novel mutation. AB - Mutations in the human melanocortin-4 receptor (MC4-R) gene may account for up to 5.8% of morbid nonsyndromic obesity. We have screened 120 unrelated obese patients for variants of the MC4-R gene. Four heterozygous missense variants were detected, including two polymorphisms (Val(103)Ile and Ile(251)Leu) previously described in the literature. A novel heterozygous mutation (Glu(308)Lys) was detected in a 36-yr-old female patient. Compared with the wild-type receptor, cells expressing the mutated receptor showed a reduced stimulation of cAMP production and a reduction of radioactive alpha MSH binding. No segregation of the mutation with the obese phenotype could be demonstrated. A second, potentially pathogenic mutation (Ser(30)Phe) was detected in a 31-yr-old female patient. Functional analysis of the mutated receptor showed no change in the affinity to the natural ligand alpha MSH nor limited ability to stimulate cAMP production. Sixty lean subjects were also screened, and no additional variants of the MC4-R gene were observed, except for two individuals with the Val(103)Ile polymorphism. In conclusion, we have screened a population of Italian obese subjects for MC4-R variants, demonstrating a 1.7% prevalence of potentially pathogenic mutations. A novel heterozygous missense mutation (Glu(308)Lys) that impairs MC4-R functional activity in vitro was characterized. PMID- 14764813 TI - Growth hormone increases vascular cell adhesion molecule 1 expression: in vivo and in vitro evidence. AB - We investigated the impact of GH administration on endothelial adhesion molecules, vascular cell adhesion molecule-1 (VCAM-1) and E-selectin, in vivo and in vitro. Soluble VCAM-1, E-selectin, and C-reactive protein concentrations were measured before and after treatment in 25 healthy subjects and 25 adult GH deficient (GHD) patients randomized to GH treatment or placebo. Furthermore, we studied the direct effect of GH and IGF-I and serum from GH-treated subjects on basal and TNF alpha-stimulated expression of VCAM-1 and E-selectin on cultured human umbilical vein endothelial cells. Baseline levels of VCAM-1, but not E selectin, were significantly lower in GHD patients than in healthy subjects (362 +/- 15 microg/liter vs. 516 +/- 21 microg/liter, P < 0.001) and increased in GHD patients during GH treatment, compared with placebo [net difference between groups 151.8 microg/liter (95% confidence interval: 95.0-208.7 microg/liter); P < 0.0001]. In human umbilical vein endothelial cells, there was no direct stimulatory effect of either GH or IGF-I on the expression of VCAM-1 and E selectin, but serum from GH-treated healthy subjects significantly increased the expression of VCAM-1 (P < 0.01). Our findings are compatible with the notion that GH may stimulate the expression of VCAM-1 indirectly through modulation of circulating factors. VCAM-1-mediated leukocyte extravasation is implicated in several illnesses including atherosclerosis and multiple-organ failure in sepsis, and we hypothesize that enhanced expression of VCAM-1 may contribute to the detrimental effects of GH in critically ill patients. PMID- 14764814 TI - Influence of SHBG gene pentanucleotide TAAAA repeat and D327N polymorphism on serum sex hormone-binding globulin concentration in hirsute women. AB - SHBG is the specific plasma transport protein for sex steroid hormones in humans. Plasma SHBG concentration follows a gender dimorphism but varies with nutritional and hormonal status in both sexes. In addition, a genetic influence on SHBG in humans has recently been suggested by family studies. We investigated the relationship between a point mutation (D327N) in SHBG gene exon 8 that delays human SHBG half-life and a pentanucleotide repeat polymorphism [PNRP (TAAAA)(n)] in the SHBG gene 5' untranslated region that influences transcription in vitro, on the one hand, and SHBG levels on the other, in a population of 303 women referred for hirsutism. Of these patients, 154 (51%) met the criteria for polycystic ovary syndrome (PCOS) and 124 (41%) were overweight [body mass index (BMI) > or = 25 kg/m(2)]. The two SHBG gene alleles for D327N substitution, wild type (W) and variant (v), were identified by restriction fragment length polymorphism in the whole population, and the GeneScan method was used to identify PNRP alleles in 245 subjects. Six alleles of the pentanucleotide motif with six to 11 repeats were present in our population. Plasma SHBG concentration was related to PCOS status, non-SHBG-bound testosterone, BMI, fasting blood glucose level, fasting insulinemia, and D327N allele v. The v allele was associated with higher SHBG levels [36.9 +/- 15.9 nmol/liter for W/v (n = 52) and 43.5 +/- 3.5 nmol/liter for v/v (n = 2)] than was the wild-type W allele [31.1 +/ 16.1 nmol/liter (n = 249); P = 0.039]. Multivariate analysis showed that BMI, PCOS status, and D327N polymorphism influenced plasma SHBG concentrations, each of these parameters contributing independently of the others. Investigating the role of each allele of the TAAAA repeat polymorphism on SHBG levels was more complex because of the number of different genotypes (as many as 18 in our population) and the low frequency of some of them. Moreover, a strong disequilibrium linkage was found between D327N allele v and the eight-TAAAA repeat allele (P < 0.0001). This could mask the effect of the TAAAA repeat polymorphism on SHBG concentration in vivo. Nevertheless, SHBG levels in patients who were homozygous for six repeats (34.9 +/- 16.2 nmol/liter; n = 21) were significantly (P = 0.043) higher than in nine-repeat homozygous patients (21.5 +/ 13.0 nmol/liter; n = 8), and lay between the two for eight-repeat homozygous patients (28.5 +/- 15.8 nmol/liter; n = 44). Delineating the precise role of this PNRP polymorphism will need further investigation in a large healthy population. In summary, although BMI and PCOS status have a major influence on circulating SHBG levels in hirsute women, the present results support the notion that polymorphism(s) within the coding sequence and, potentially, in the regulatory sequence of the SHBG gene are associated with circulating SHBG levels and may represent part of the genetic background of sex steroid hormone activity in humans. PMID- 14764815 TI - Acute insulin responses to calcium and tolbutamide do not differentiate focal from diffuse congenital hyperinsulinism. AB - Congenital hyperinsulinism (CHI) is related to two main histological pancreas anomalies: focal adenomatous hyperplasia and diffuse beta-cell hypersecretion. Pharmacological tests to measure acute insulin responses (AIR) to peripheral i.v. injections of glucose, calcium, and tolbutamide have been reported as potential means to distinguish between these histological forms. In patients with defects in ATP-sensitive potassium channels, tolbutamide will fail to induce insulin release in affected portions of the pancreas, whereas calcium gluconate will enhance insulin release through spontaneously active voltage-gated Ca(2+) channels. Consequently, in focal CHI patients, calcium should promote AIRs from the lesion, whereas tolbutamide should act to promote insulin secretion from the healthy region of the pancreas (outside the focal hyperplasia). We therefore studied AIRs to calcium and tolbutamide stimulation tests in 16 children with focal (n = 9) or diffuse (n = 7) CHI before pancreatic surgery. We found hypervariable AIRs to glucose and calcium stimulation in both focal and diffuse CHI patients. AIRs to tolbutamide stimulation were found modest in focal CHI patients, which might account for beta-cell quiescence in the healthy portion of the pancreas of these patients. We conclude that AIRs to calcium and tolbutamide stimulation tests are not sufficient to differentiate the focal from the diffuse CHI patients. PMID- 14764816 TI - Evidence for enhanced adipogenesis in the orbits of patients with Graves' ophthalmopathy. AB - The signs and symptoms of Graves' ophthalmopathy (GO) result from increased volume of the orbital contents, including adipose, connective, and extraocular muscle tissues. We wanted to determine whether the expanded adipose tissue volume might be in part attributable to de novo adipogenesis. We measured levels of mRNA encoding leptin, adiponectin, peroxisome proliferator-activated receptor gamma (PPAR gamma), preadipocyte factor-1, and TSH receptor (TSHr) genes in orbital adipose tissues from GO patients (n = 22) and normal individuals (n = 18) and in orbital preadipocyte cultures derived from GO patients (n = 6) and normal subjects (n = 3) using quantitative real-time RT PCR. We found increased leptin, adiponectin, PPAR gamma, and TSHr expression in GO compared with normal orbital tissue samples, with positive correlations in the GO tissues between TSHr and leptin, adiponectin and PPAR gamma. In vitro differentiation of GO and normal preadipocytes resulted in enhanced adiponectin, leptin, and TSHr expression, with greater expression of the latter two genes in the GO cultures. These results suggest that de novo adipogenesis within orbital tissues with parallel enhanced expression of TSHr may be important in the pathogenesis of GO, and that potential therapies for GO might include inhibition of the adipogenic pathway. PMID- 14764817 TI - The relationship between active ghrelin levels and human obesity involves alterations in resting energy expenditure. AB - Ghrelin is a gastric hormone that exerts a stimulatory effect on appetite and fat accumulation. Ser(3) octanoylation is regarded as a prerequisite for ghrelin biological activity, although des-octanoylated forms may retain biological functions in vitro. Circulating ghrelin levels are usually low in obesity and in states of positive energy balance. Hence, the aim of our study was to analyze plasma active and serum total ghrelin levels in 20 obese (ages, 22-42 yr; body mass index, 41.3 +/- 1.1 kg/m(2)) and 20 lean subjects (ages, 22-43 yr; body mass index, 22.4 +/- 0.6 kg/m(2)) as well as their relationship to measures of glucose homeostasis, body fat, and resting energy expenditure (REE). The measured/predicted REE percentage ratio was calculated to subdivide groups into those with positive (> or = 100% ) and negative (<100%) ratio values. In obese patients, plasma active (180 +/- 18 vs. 411 +/- 57 pg/ml; P < 0.001) and serum total ghrelin levels (3650 +/- 408 vs. 5263 +/- 643 pg/ml; P < 0.05) were significantly lower when compared with lean subjects. Hence, ghrelin activity, defined as the proportion of active over total ghrelin levels, was similarly reduced in the obese state (6.1 +/- 0.9% vs. 8.4 +/- 1%; P < 0.05). There was a significant correlation between active and total ghrelin (r = 0.62; P < 0.001), and between total ghrelin and insulin (r = -0.53; P < 0.001) or insulin resistance using the homeostatis model of assessment-insulin resistance (r = 0.49; P < 0.001) approach. Significantly higher active ghrelin levels (214 +/- 22 vs. 159 +/- 30 pg/ml; P < 0.05) and ghrelin activity (8 +/- 1.7% vs. 4.9 +/- 0.9%; P < 0.05) were observed in patients with positive compared with negative measured/predicted REE ratio values. Our study shows that obesity is associated with an impairment of the entire ghrelin system. The observation that ghrelin is further decreased in cases of abnormal energy profit adds new evidence to the relationship between ghrelin activity and energy balance in obesity. PMID- 14764818 TI - Identification and characterization of melanocortin-4 receptor gene mutations in morbidly obese finnish children and adults. AB - Two Finnish cohorts, comprising 56 children with severe early-onset obesity (relative weight for height greater than or equal to +70% before age 10) and 252 morbidly obese adults (body mass index, > or = 40 kg/m(2)), were screened for melanocortin-4 receptor (MC4R) mutations. We identified a pathogenic mutation (S127L) in one child, causing severe early-onset obesity. We describe the phenotype of this particular mutation for the first time. We also identified a novel (I226T) polymorphism in the coding and two new variations (-439delGC and 1059C>T) outside the coding region of the MC4R gene. Three previously described polymorphisms (V103I, T112M, and I125L) were identified. In vitro functional studies of variants T112M, S127L, and I226T supported a pathogenic role of the S127L mutation, because signaling properties of the receptor in response to the MC4R agonists alpha-MSH, beta-MSH, and gamma(1)-MSH were impaired. The S127L mutation did not affect receptor inhibition by the antagonist agouti-related protein. Localization of the three variant receptors was similar to that of wild type. In conclusion, a pathogenic MC4R mutation was found among subjects with severe early-onset obesity but not among morbidly obese adults. Impaired function of the S127L receptor was due to reduced activation, not a defect of protein transport to the cell membrane. PMID- 14764819 TI - Congenital lipoid adrenal hyperplasia caused by a novel splicing mutation in the gene for the steroidogenic acute regulatory protein. AB - Steroidogenic acute regulatory protein (StAR) plays a crucial role in the transport of cholesterol from the cytoplasm to the inner mitochondrial membrane, facilitating its conversion to pregnenolone by cytochrome P450scc. Its essential role in steroidogenesis was demonstrated after observing that StAR gene mutations gave rise to a potentially lethal disease named congenital lipoid adrenal hyperplasia, in which virtually no steroids are produced. We report here a 2 month-old female patient, karyotype 46XY, who presented with growth failure, convulsions, dehydration, hypoglycemia, hyponatremia, hypotension, and severe hyperpigmentation suggestive of adrenal insufficiency. Serum cortisol, 17OH progesterone, dehydroepiandrosterone sulfate, testosterone, 17OH-pregnenolone, and aldosterone levels were undetectable in the presence of high ACTH and plasma renin activity levels. Immunohistochemical analysis of testis tissues revealed the absence of StAR protein. Molecular analysis of StAR gene demonstrated a homozygous G to T mutation within the splice donor site of exon 1 (IVS1 + 1G>T). Her parents and one brother were heterozygous for this mutation. In vitro analysis of the mutation was performed in COS cells transfected with minigenes coding regions spanning exon-intron 1 to 3 carrying the mutant and the wild-type sequences. RT-PCR analyses of the mutant gene showed an abnormal mRNA transcript of 2430 bp (normal size 433 bp). Sequence analysis of the mutant mRNA demonstrated the retention of intron 1. Immunolocalization of the StAR minigene product detected the peptide in the mitochondria of COS cells transfected with the wild-type minigene but not in those transfected with the mutant minigene. We conclude that this mutation gives rise to a truncated StAR protein, which lacks an important N-terminal region and the entire lipid transfer domain. PMID- 14764820 TI - Human fetal nongonadal tissues contain human chorionic gonadotropin/luteinizing hormone receptors. AB - Human chorionic gonadotropin (hCG), a heterodimeric glycoprotein hormone produced in abundance by placental syncytiotrophoblasts, is preferentially secreted into maternal circulation. Fetal circulation also contains low levels of hCG that are probably derived from fetal kidney, liver, anterior pituitary gland, etc. In addition, the fetus has access to hCG present in exocoelomic and amniotic fluids. hCG has been found in a number of fetal tissues known to stimulate fetal adrenal and testicular steroidogenesis and is also thought to play a role in growth and differentiation of fetal tissues. This led us to test the hypothesis that fetal nongonadal tissues, as in the adult, may also contain hCG/LH receptors. This hypothesis was tested by immunocytochemistry, Western blotting, in situ hybridization, and RT-PCR. The results demonstrate that kidney, liver, pancreas, lung, small and large intestines, and adrenals contained hCG/LH receptors. Although the role of fetal nongonadal hCG/LH receptors is not known, they may mediate the pleiotropic actions of hCG in the growing human fetus. PMID- 14764821 TI - Molecular study of the 3 beta-hydroxysteroid dehydrogenase gene type II in patients with hypospadias. AB - To determine whether some patients with idiopathic hypospadias have HSD3B2 mutations, we genotyped this locus in 90 patients with hypospadias (age, 6.0 +/- 0.4 yr) and 101 healthy fertile male controls. We measured basal plasma renin activity and performed an ACTH test for determination of 17-OH-pregnenolone, 17 OH-progesterone, cortisol, dehydroepiandrosterone sulfate, and androstenedione and an human chorionic gonadotropin test for determination of androstenedione, testosterone, and dihydrotestosterone. We did not observe a clear steroidogenic pattern suggestive of 3 beta-HSD deficiency in any patient. DNA was extracted from peripheral lymphocytes; and exons 1, 2, 3, and 4 were amplified by PCR and analyzed by denaturing gradient gel electrophoresis. An abnormal electrophoretic migration pattern of exon 4 was observed in five patients. Two patients had missense heterozygous mutations (S213T and S284R). In another three patients, we observed heterozygous nucleotide variants in exon 4 that did not produce a change in amino acids (A238, T259, T320). In vitro enzymatic activity was diminished by 40% and 32% in the S213T and S284R heterozygous mutations, respectively. One control exhibited a heterozygous mutation in exon 3 (V78I), which did not alter in vitro enzyme activity. In addition, we observed possible polymorphisms in intron 1 in four patients and one control. We conclude that subtle molecular abnormalities in the HSD3B2 gene may be observed in some patients with apparent idiopathic hypospadias but that this finding is uncommon. PMID- 14764822 TI - Corticotropin-releasing hormone system in human adipose tissue. AB - Mounting evidence exists for a role of the CRH system in energy balance, including a direct influence on human adipocytes, the regulation of adipose 11 beta-hydroxysteroid dehydrogenase type 1 activity, and cortisol formation. We characterized the expression of CRH receptors 1 and 2 and CRH-like peptides stresscopin and urocortin in human adipose tissue in comparison with other peripheral tissues, adrenal, and heart. The effect of CRH on CRH receptor and CRH like peptide expression was analyzed in isolated human adipocytes using quantitative TaqMan PCR. CRH receptors were detectable in fat tissue at mRNA and protein levels. CRH-R2 expression in fat was comparable with its expression in the heart, the organ with the highest CRH-R2 expression known. CRH-R1:CRH-R2 ratio varied according to fat-depot type; whereas CRH-R1 expression was higher in sc fat than in visceral fat, the opposite was true for CRH-R2. Adipose tissue also expressed urocortin and stresscopin, the predominant ligands of peripheral CRH-R2. CRH down-regulated CRH-R1 and CRH-R2 mRNA expression in isolated adipocytes. These data, together with the recently published observation that CRH regulates adipocyte metabolism by down-regulating 11 beta-hydroxysteroid dehydrogenase, indicate that a CRH system exists within human adipose tissue. This system could be implicated in energy homeostasis and in mediating the anorexic effects of CRH at adipose level. PMID- 14764823 TI - Insulin promoter factor-1 mutations and diabetes in Trinidad: identification of a novel diabetes-associated mutation (E224K) in an Indo-Trinidadian family. AB - This study investigated the prevalence of insulin promoter factor-1(IPF-1) mutations in familial early-onset diabetes mellitus in Trinidad. We screened 264 unrelated subjects with type 2 diabetes diagnosed before 40 yr of age and a family history of diabetes for mutations in the minimal promoter and coding region of the IPF-1 gene (IPF1). This study population included 169 patients of East Indian descent (Indo-Trinidadians), 66 of African descent (Afro Trinidadians), and 29 of mixed ancestry. We identified five IPF1 variants, including one new missense mutation E224K, the previously described diabetes associated duplication P242 P243dupP, two silent mutations in the codons for Leu54 (c.162G>A) and Ala256 (c.768C>A), and a substitution in the 5'-untranslated region (c.-18C>T). The E224K mutation was found in two unrelated diabetic Indo Trinidadians and 0 of 60 controls. It was present on the same haplotype in both patients suggesting a founder effect. The E224K mutation cosegregated with early onset diabetes or impaired glucose tolerance in a large family, suggestive of the type 4 form of maturity-onset diabetes of the young rather than type 2 diabetes. Functional studies of E224K showed reduced transactivation activity. IPF1 mutations leading to synthesis of a mutant protein may contribute to the development of familial early-onset diabetes/maturity-onset diabetes of the young in Indo-Trinidadians. PMID- 14764825 TI - Expression, localization, and signaling of prostaglandin F2 alpha receptor in human endometrial adenocarcinoma: regulation of proliferation by activation of the epidermal growth factor receptor and mitogen-activated protein kinase signaling pathways. AB - Prostaglandin F(2 alpha)(PGF(2 alpha)) is a bioactive lipid biosynthesized by cyclooxygenase (COX) enzymes and mediates its biological activity via the heptahelical G(q)-coupled PGF(2 alpha)receptor (FP receptor). This study investigated the expression and molecular signaling of the FP receptor in human endometrial adenocarcinomas. Real-time RT-PCR and Western blot analysis confirmed FP receptor expression in endometrial adenocarcinoma of all grades and differentiation. The expression of FP receptor was up-regulated in all endometrial adenocarcinomas compared with normal endometrium. The site of FP receptor expression was localized by in situ hybridization and immunohistochemistry to the neoplastic epithelial cells in all adenocarcinomas. Treatment of endometrial adenocarcinoma explants with PGF(2 alpha) resulted in mobilization of inositol phosphate signaling, indicating functional FP receptor expression. We investigated whether PGF(2 alpha) could trans-activate the epidermal growth factor receptor (EGFR) and trigger the MAPK signaling pathway. Treatment of adenocarcinoma explants and endometrial adenocarcinoma cells (Ishikawa) with PGF(2 alpha)-phosphorylated EGFR, triggered MAPK signaling and enhanced the proliferation of Ishikawa cells. Inactivation of phospholipase C, EGFR kinase, and MAPK kinase with specific inhibitors abolished PGF(2 alpha) induced trans-activation of EGFR, MAPK signaling, and Ishikawa cell proliferation. These data suggest that PGF(2 alpha)-FP receptor promote endometrial tumorigenesis via a phospholipase C-mediated phosphorylation of the EGFR and MAPK signaling pathways. PMID- 14764824 TI - Enhanced lipoprotein lipase secretion and foam cell formation by macrophages of patients with growth hormone deficiency: possible contribution to increased risk of atherogenesis? AB - GH deficiency is associated with increased prevalence of atherosclerosis, and recent data indicate a proatherogenic role for macrophage lipoprotein lipase (LPL) in the arterial wall. In this pilot study, we determined LPL expression and foam cell formation in monocyte-derived macrophages of 12 control subjects and nine patients with GH deficiency without GH replacement therapy. LPL mRNA levels, mass, and activity were increased in macrophages of patients with GH deficiency. In these subjects, macrophage LPL activity correlated with body mass index and fat mass. Incubation of patient macrophages with IGF-I for 24 h or differentiation of monocytes isolated from GH-deficient patients into macrophages in the presence of this growth factor decreased the amount of LPL mass. Compared with control cells, macrophages derived from GH-deficient patients took up and stored increased amounts of proatherogenic lipoproteins and were more easily converted to foam cells. In the supernatants of these cells, increased levels of free fatty acids and TNFalpha were also documented. These results demonstrate that macrophages of patients with GH deficiency secrete increased amounts of proatherogenic cytokines and are more susceptible to foam cell formation. These alterations may contribute to the increased cardiovascular risk in patients with GH deficiency. PMID- 14764826 TI - Transcriptional profiling reveals coordinated up-regulation of oxidative metabolism genes in thyroid oncocytic tumors. AB - Oncocytomas are large cell tumors characterized by an abnormal proliferation of mitochondria. To investigate this phenomenon in thyroid oncocytomas, we determined gene expression profiles of 87 samples using microarrays of 6720 PCR products from cDNA clones. Samples included 29 thyroid oncocytomas and six papillary carcinomas, the remainder representing other thyroid pathologies or mitochondria-rich tumor samples, normal thyroid samples, and two thyroid cell lines. Hierarchical clustering and supervised analysis identified two specific oncocytic clusters and 163 distinctly regulated genes between oncocytoma and normal thyroid. Differential expression of five selected genes (APOD, BCL-2, COX, CTSB, and MAP2) was confirmed by immunohistochemistry. The two specific oncocytic clusters were rich in mitochondrial genes and revealed coordinated expression of nuclear and mitochondrial respiratory chain genes. We also observed the up regulation of genes involved in mitochondrial biogenesis, such as nuclear respiratory factor 1 and the endothelial nitric oxide synthase. Several oxidative metabolism genes were overexpressed in oncocytomas, including those from the tricarboxylic acid cycle (MDH1) and cytosolic glycolysis (GAPD, ENO1, and GPI). On the contrary, the lactate dehydrogenase A gene, involved in anaerobic metabolism, was down-regulated. Our results suggest that, unlike a large number of solid tumors, thyroid oncocytomas produce energy through an aerobic pathway. PMID- 14764827 TI - Valproic acid induces the expression of the Na+/I- symporter and iodine uptake in poorly differentiated thyroid cancer cells. AB - In poorly differentiated thyroid cancer, molecular characteristics are reported to be lost such as to cause insensitivity of the tumor to radiometabolic therapy. Considerable work is in progress to identify compounds that redifferentiate thyroid cancer cells. The present study evaluates the action of valproic acid, a potent anticonvulsant recently reported to inhibit histone deaceytlase, on cultured thyroid cancer cells. N-PA (poorly differentiated) and ARO (anaplastic) cells were treated with increasing valproic acid concentrations.; expression of mRNA and cell localization pattern for the Na+/I- symporter (NIS), as well as 125I uptake, were evaluated before and after treatment. Valproic acid induced NIS gene expression, NIS membrane localization and iodide accumulation in N-PA cells; it was effective at clinically-safe doses in the therapeutic range. In ARO cells, only induction of NIS mRNA was observed, and was not followed by any change in iodide uptake. Valproic acid is thus effective at restoring the ability of N-PA cells to accumulate iodide and its use in clinical trials may be recommended. PMID- 14764828 TI - Prostaglandins differentially modulate progesterone receptor-A and -B expression in human myometrial cells: evidence for prostaglandin-induced functional progesterone withdrawal. AB - We tested the hypothesis that prostaglandin (PGs), PGE2, and PGF2 alpha, stimulate labor and delivery in women, in part, by inducing functional progesterone withdrawal in myometrial cells by increasing the progesterone receptor (PR)-A/PR-B expression ration. PHM1-31 cells (an immortal pregnant human myometrial cell line) were exposed to PGE2, PGF2 alpha, cyclic-8-bromoadenosine monophosphate (8-Br-cAMP) and phorbol 12-myristate 13-acetate (PMA) at various concentrations for 24h. Effects on PR-A and PR-B expression were then assessed by quantitative RT-PCR. PGF2 alpha dose dependently increased PR-A mRNA and the PR A/PR-B expression ration but did not effect PR-B mRNA. PGE2 dose-dependently increased mRNAs encoding PR-A and PR-B. The PGE2 dose-threshold for PR-A (0.01 nM) was lower than that for PR-B (0.1 nM), which resulted in an initial rise then a gradual fall in PR-A/PR-B expression ration to basal levels in response to PGE2. Activation of the protein kinase (PK)-A signaling pathway with 8-Br-cAMP coordinately increased expression of PR-A and PR-B and therefore did not alter the PR-A/PR-B expression ration. In contrast, activation of the PKC signaling pathway with PMA increased expression of PR-A without affecting PR-B and therefore significantly (P<0.05) increased the PR-A/PR-B expression ration. These data demonstrate differential control of myometrial PR-A and PR-B expression by PGE2 and PGF2 alpha and by specific intracellular signaling pathways. We conclude that PGs acting via the PKC pathway facilitate functional progesterone withdrawal by increasing the myometrial PR-A/PR-B expression ratio. PMID- 14764829 TI - Author's response: The endocrine care of transsexual people. PMID- 14764830 TI - The endocrine care of transsexual people. PMID- 14764832 TI - The effect of octreotide on parathyroid carcinoma. PMID- 14764833 TI - Science at the leading edge. PMID- 14764835 TI - France. Researchers issue an ultimatum. PMID- 14764834 TI - Nuclear physics. New chemical elements probe the shoals of stability. PMID- 14764836 TI - Biomedical politics. Sex studies 'properly' approved. PMID- 14764837 TI - Condensed-matter physics. New-style matter opens cool middle ground. PMID- 14764838 TI - South Asia. India, Pakistan hold first science talks. PMID- 14764839 TI - Intellectual property. Court tells Nichia to pay blue LED inventor $180 million. PMID- 14764840 TI - Genetics. Development gene may give nerve cells a sense of identity. PMID- 14764841 TI - Psychopharmacology. FDA weighs suicide risk in children on antidepressants. PMID- 14764842 TI - Space-based astronomy. Hubble huggers get a reprieve. PMID- 14764843 TI - Nation building. Reports to U.N. propose bigger role for science. PMID- 14764844 TI - Science policy. 2005 budget makes flat a virtue. PMID- 14764845 TI - Science policy. Highlights from the budget. PMID- 14764846 TI - John Delaney profile. Marine geologist hopes to hear the heartbeat of the planet. PMID- 14764847 TI - John Delaney profile. A cautionary tale From Bermuda. PMID- 14764848 TI - DNA forensics. Buried, recovered, lost again? The Romanovs may never rest. PMID- 14764849 TI - Robert Aymar interview. The man to finish the job. Interview by Daniel Clery. PMID- 14764850 TI - Robert Aymar interview. A leviathan takes shape beneath Geneva's gentle environs. PMID- 14764852 TI - An inhospitable environment for women? PMID- 14764851 TI - More on mercury content in fish. PMID- 14764855 TI - Beyond the ivory tower. The sinews of war: ancient catapults. PMID- 14764853 TI - Comment on "Tubular graphite cones". PMID- 14764856 TI - Cell biology. Integrins, rafts, Rac, and Rho. PMID- 14764857 TI - Genetics. Robust interactions. PMID- 14764858 TI - Cancer. Respect thy neighbor! PMID- 14764859 TI - Materials science. Thermoelectricity in semiconductor nanostructures. PMID- 14764860 TI - Physics. Pas de deux for atomic electrons. PMID- 14764862 TI - Life's patterns: no need to spell it out? PMID- 14764863 TI - The new math of clinical trials. PMID- 14764864 TI - Making sense of a heart gone wild. PMID- 14764865 TI - Introductory science and mathematics education for 21st-Century biologists. AB - Galileo wrote that "the book of nature is written in the language of mathematics"; his quantitative approach to understanding the natural world arguably marks the beginning of modern science. Nearly 400 years later, the fragmented teaching of science in our universities still leaves biology outside the quantitative and mathematical culture that has come to define the physical sciences and engineering. This strikes us as particularly inopportune at a time when opportunities for quantitative thinking about biological systems are exploding. We propose that a way out of this dilemma is a unified introductory science curriculum that fully incorporates mathematics and quantitative thinking. PMID- 14764866 TI - Uses and abuses of mathematics in biology. AB - In the physical sciences, mathematical theory and experimental investigation have always marched together. Mathematics has been less intrusive in the life sciences, possibly because they have until recently been largely descriptive, lacking the invariance principles and fundamental natural constants of physics. Increasingly in recent decades, however, mathematics has become pervasive in biology, taking many different forms: statistics in experimental design; pattern seeking in bioinformatics; models in evolution, ecology, and epidemiology; and much else. I offer an opinionated overview of such uses--and abuses. PMID- 14764867 TI - Evolutionary dynamics of biological games. AB - Darwinian dynamics based on mutation and selection form the core of mathematical models for adaptation and coevolution of biological populations. The evolutionary outcome is often not a fitness-maximizing equilibrium but can include oscillations and chaos. For studying frequency-dependent selection, game theoretic arguments are more appropriate than optimization algorithms. Replicator and adaptive dynamics describe short- and long-term evolution in phenotype space and have found applications ranging from animal behavior and ecology to speciation, macroevolution, and human language. Evolutionary game theory is an essential component of a mathematical and computational approach to biology. PMID- 14764868 TI - Inferring cellular networks using probabilistic graphical models. AB - High-throughput genome-wide molecular assays, which probe cellular networks from different perspectives, have become central to molecular biology. Probabilistic graphical models are useful for extracting meaningful biological insights from the resulting data sets. These models provide a concise representation of complex cellular networks by composing simpler submodels. Procedures based on well understood principles for inferring such models from data facilitate a model based methodology for analysis and discovery. This methodology and its capabilities are illustrated by several recent applications to gene expression data. PMID- 14764869 TI - A SNP map of the rat genome generated from cDNA sequences. PMID- 14764871 TI - Probing two-electron dynamics of an atom. AB - Coherent short-pulse laser excitation has been used to control the approximate energy and relative proximity of two valence electrons within the same alkaline earth atom, thereby providing insight into the dynamical evolution of a three body Coulomb system. Our time-domain experiments enable direct experimental study of the electron dynamics at the classical limit of a two-electron atom. As an example, we look at the mechanism of autoionization for one two-electron configuration class and find that the doubly excited atom decays through a single violent electron-electron collision rather than a gradual exchange of energy between the electrons. PMID- 14764872 TI - Profiling the thermoelectric power of semiconductor junctions with nanometer resolution. AB - We have probed the local thermoelectric power of semiconductor nanostructures with the use of ultrahigh-vacuum scanning thermoelectric microscopy. When applied to a p-n junction, this method reveals that the thermoelectric power changes its sign abruptly within 2 nanometers across the junction. Because thermoelectric power correlates with electronic structure, we can profile with nanometer spatial resolution the thermoelectric power, band structures, and carrier concentrations of semiconductor junctions that constitute the building blocks of thermoelectric, electronic, and optoelectronic devices. PMID- 14764870 TI - Global mapping of the yeast genetic interaction network. AB - A genetic interaction network containing approximately 1000 genes and approximately 4000 interactions was mapped by crossing mutations in 132 different query genes into a set of approximately 4700 viable gene yeast deletion mutants and scoring the double mutant progeny for fitness defects. Network connectivity was predictive of function because interactions often occurred among functionally related genes, and similar patterns of interactions tended to identify components of the same pathway. The genetic network exhibited dense local neighborhoods; therefore, the position of a gene on a partially mapped network is predictive of other genetic interactions. Because digenic interactions are common in yeast, similar networks may underlie the complex genetics associated with inherited phenotypes in other organisms. PMID- 14764873 TI - Cubic AgPb(m)SbTe(2+m): bulk thermoelectric materials with high figure of merit. AB - The conversion of heat to electricity by thermoelectric devices may play a key role in the future for energy production and utilization. However, in order to meet that role, more efficient thermoelectric materials are needed that are suitable for high-temperature applications. We show that the material system AgPb(m)SbTe(2+m) may be suitable for this purpose. With m = 10 and 18 and doped appropriately, n-type semiconductors can be produced that exhibit a high thermoelectric figure of merit material ZTmax of approximately 2.2 at 800 kelvin. In the temperature range 600 to 900 kelvin, the AgPb(m)SbTe(2+m) material is expected to outperform all reported bulk thermoelectrics, thereby earmarking it as a material system for potential use in efficient thermoelectric power generation from heat sources. PMID- 14764874 TI - Superlattices of iron nanocubes synthesized from Fe[N(SiMe3)2]2. AB - The reaction of the metal-organic precursor Fe[N(SiMe3)2]2 with H2 in the presence of a long-chain acid and a long-chain amine in various proportions produces monodisperse zerovalent iron nanoparticles. These Fe particles display magnetic properties that match those of bulk iron as evidenced by magnetic and Mossbauer measurements. The nanoparticles adopt a cubic shape with edges of 7 nanometers and are incorporated into extended crystalline superlattices containing nanocubes in close proximity and with their crystallographic axes aligned. These superlattices are formed in solution, precipitate in high yield, and may be redissolved and redeposited as two-dimensional arrays. PMID- 14764875 TI - Rheology and microscopic topology of entangled polymeric liquids. AB - The viscoelastic properties of high molecular weight polymeric liquids are dominated by topological constraints on a molecular scale. In a manner similar to that of entangled ropes, polymer chains can slide past but not through each other. Tube models of polymer dynamics and rheology are based on the idea that entanglements confine a chain to small fluctuations around a primitive path that follows the coarse-grained chain contour. Here we provide a microscopic foundation for these highly successful phenomenological models. We analyze the topological state of polymeric liquids in terms of primitive paths and obtain parameter-free, quantitative predictions for the plateau modulus, which agree with experiment for all major classes of synthetic polymers. PMID- 14764876 TI - 48,000 years of climate and forest change in a biodiversity hot spot. AB - A continuous 48,000-year-long paleoecological record from Neotropical lower montane forest reveals a consistent forest presence and an ice-age cooling of approximately 5 degrees to 9 degrees C. After 30,000 years of compositional stability, a steady turnover of species marks the 8000-year-long transition from ice-age to Holocene conditions. Although the changes were directional, the rates of community change were no different during this transitional period than in the preceding 30,000-year period of community stability. The warming rate of about 1 degrees C per millennium during the Pleistocene-Holocene transition was an order of magnitude less than the projected changes for the 21st century. PMID- 14764877 TI - Role of microbes in the smectite-to-illite reaction. AB - Temperature, pressure, and time have been thought to control the smectite-to illite (S-I) reaction, an important diagenetic process used for petroleum exploration. We demonstrated that microorganisms can promote the S-I reaction by dissolving smectite through reduction of structural Fe(III) at room temperature and 1 atmosphere within 14 days. This reaction typically requires conditions of 300 degrees to 350 degrees C, 100 megapascals, and 4 to 5 months in the absence of microbial activity. These results challenge the conventional concept of the S I reaction and of reaction kinetic models. PMID- 14764878 TI - Genome-wide RNAi analysis of growth and viability in Drosophila cells. AB - A crucial aim upon completion of whole genome sequences is the functional analysis of all predicted genes. We have applied a high-throughput RNA interference (RNAi) screen of 19,470 double-stranded (ds) RNAs in cultured cells to characterize the function of nearly all (91%) predicted Drosophila genes in cell growth and viability. We found 438 dsRNAs that identified essential genes, among which 80% lacked mutant alleles. A quantitative assay of cell number was applied to identify genes of known and uncharacterized functions. In particular, we demonstrate a role for the homolog of a mammalian acute myeloid leukemia gene (AML1) in cell survival. Such a systematic screen for cell phenotypes, such as cell viability, can thus be effective in characterizing functionally related genes on a genome-wide scale. PMID- 14764879 TI - Localized stabilization of microtubules by integrin- and FAK-facilitated Rho signaling. AB - Microtubule (MT) stabilization is regulated by the small guanosine triphosphate (GTP)-binding protein Rho and its effector, mammalian homolog of Diaphanous (mDia), in migrating cells, but factors responsible for localized stabilization at the leading edge are unknown. We report that integrin-mediated activation of focal adhesion kinase (FAK) at the leading edge is required for MT stabilization by the Rho-mDia signaling pathway in mouse fibroblasts. MT stabilization also involved FAK-regulated localization of a lipid raft marker, ganglioside GM1, to the leading edge. The integrin-FAK signaling pathway may facilitate Rho-mDia signaling through GM1, or through a specialized membrane domain containing GM1, to stabilize MTs in the leading edge of migrating cells. PMID- 14764880 TI - Integrins regulate Rac targeting by internalization of membrane domains. AB - Translocation of the small GTP-binding protein Rac1 to the cell plasma membrane is essential for activating downstream effectors and requires integrin-mediated adhesion of cells to extracellular matrix. We report that active Rac1 binds preferentially to low-density, cholesterol-rich membranes, and specificity is determined at least in part by membrane lipids. Cell detachment triggered internalization of plasma membrane cholesterol and lipid raft markers. Preventing internalization maintained Rac1 membrane targeting and effector activation in nonadherent cells. Regulation of lipid rafts by integrin signals may regulate the location of membrane domains such as lipid rafts and thereby control domain specific signaling events in anchorage-dependent cells. PMID- 14764881 TI - Large shifts in pathogen virulence relate to host population structure. AB - Theory on the evolution of virulence generally predicts selection for an optimal level of virulence determined by trade-offs with transmission and/or recovery. Here we consider the evolution of pathogen virulence in hosts who acquire long lived immunity and live in a spatially structured population. We show theoretically that large shifts in virulence may occur in pathogen populations as a result of a bistability in evolutionary dynamics caused by the local contact or social population structure of the host. This model provides an explanation for the rapid emergence of the highly virulent strains of rabbit hemorrhagic disease virus. PMID- 14764882 TI - TGF-beta signaling in fibroblasts modulates the oncogenic potential of adjacent epithelia. AB - Stromal cells can have a significant impact on the carcinogenic process in adjacent epithelia. The role of transforming growth factor-beta (TGF-beta) signaling in such epithelial-mesenchymal interactions was determined by conditional inactivation of the TGF-beta type II receptor gene in mouse fibroblasts (Tgfbr2fspKO). The loss of TGF-beta responsiveness in fibroblasts resulted in intraepithelial neoplasia in prostate and invasive squamous cell carcinoma of the forestomach, both associated with an increased abundance of stromal cells. Activation of paracrine hepatocyte growth factor (HGF) signaling was identified as one possible mechanism for stimulation of epithelial proliferation. Thus, TGF-beta signaling in fibroblasts modulates the growth and oncogenic potential of adjacent epithelia in selected tissues. PMID- 14764883 TI - Extracellular replication of Listeria monocytogenes in the murine gall bladder. AB - The bacterium Listeria monocytogenes can cause a life-threatening systemic illness in humans. Despite decades of progress in animal models of listeriosis, much remains unknown about the processes of infection and colonization. Here, we report that L. monocytogenes can replicate in the murine gall bladder and provide evidence that its replication there is extracellular and intraluminal. In vivo bioluminescence imaging was employed to determine the location of the infection over time in live animals, revealing strong signals from the gall bladder over a period of several days, in diseased as well as asymptomatic animals. The data suggest that L. monocytogenes may be carried in the human gall bladder. PMID- 14764884 TI - Selective D2 receptor actions on the functional circuitry of working memory. AB - Prefrontal neurons engaged by working memory tasks express a sequence of phasic and tonic activations linked to a train of sensory, mnemonic, and response related events. Here, we report that the dopamine D2 receptor selectively modulates the neural activities associated with memory-guided saccades in oculomotor delayed-response tasks yet has little or no effect on the persistent mnemonic-related activity, which is instead modulated by D1 receptors. This associates the D2 receptor with a specific component of working memory circuitry and fractionates the modulatory effects of D1 and D2 receptors on the neural machinery of a cognitive process. PMID- 14764885 TI - Architecture of the photosynthetic oxygen-evolving center. AB - Photosynthesis uses light energy to drive the oxidation of water at an oxygen evolving catalytic site within photosystem II (PSII). We report the structure of PSII of the cyanobacterium Thermosynechococcus elongatus at 3.5 angstrom resolution. We have assigned most of the amino acid residues of this 650 kilodalton dimeric multisubunit complex and refined the structure to reveal its molecular architecture. Consequently, we are able to describe details of the binding sites for cofactors and propose a structure of the oxygen-evolving center (OEC). The data strongly suggest that the OEC contains a cubane-like Mn3CaO4 cluster linked to a fourth Mn by a mono-micro-oxo bridge. The details of the surrounding coordination sphere of the metal cluster and the implications for a possible oxygen-evolving mechanism are discussed. PMID- 14764886 TI - The structure and receptor binding properties of the 1918 influenza hemagglutinin. AB - The 1918 influenza pandemic resulted in about 20 million deaths. This enormous impact, coupled with renewed interest in emerging infections, makes characterization of the virus involved a priority. Receptor binding, the initial event in virus infection, is a major determinant of virus transmissibility that, for influenza viruses, is mediated by the hemagglutinin (HA) membrane glycoprotein. We have determined the crystal structures of the HA from the 1918 virus and two closely related HAs in complex with receptor analogs. They explain how the 1918 HA, while retaining receptor binding site amino acids characteristic of an avian precursor HA, is able to bind human receptors and how, as a consequence, the virus was able to spread in the human population. PMID- 14764887 TI - Structure of the uncleaved human H1 hemagglutinin from the extinct 1918 influenza virus. AB - The 1918 "Spanish" influenza pandemic represents the largest recorded outbreak of any infectious disease. The crystal structure of the uncleaved precursor of the major surface antigen of the extinct 1918 virus was determined at 3.0 angstrom resolution after reassembly of the hemagglutinin gene from viral RNA fragments preserved in 1918 formalin-fixed lung tissues. A narrow avian-like receptor binding site, two previously unobserved histidine patches, and a less exposed surface loop at the cleavage site that activates viral membrane fusion reveal structural features primarily found in avian viruses, which may have contributed to the extraordinarily high infectivity and mortality rates observed during 1918. PMID- 14764888 TI - Intraoperative radiation therapy in liver tissue in a pig model: monitoring with dual-modality PET/CT. AB - PURPOSE: To assess, in a pig model, the value of dual-modality positron emission tomography (PET)/computed tomography (CT) for monitoring radiation therapy. MATERIALS AND METHODS: Central bile duct resection followed by creation of a biliodigestive anastomosis was performed in nine pigs. Six of these pigs were also treated with intraoperative radiation therapy (IORT) (20 Gy) in the area of the anastomosis. Two, 4, and 8 weeks postoperatively, contrast material-enhanced fluorine 18 fluorodeoxyglucose (FDG) PET/CT of the liver was performed in all of the animals. The radioactive tracer concentration in the region of the anastomosis was quantified, and the values were compared intraindividually with the values at the liver periphery. Histologic evaluation of the liver was performed 8 weeks postoperatively. The PET/CT images were assessed for changes in liver volume and bile duct diameter over time. RESULTS: In all nine pigs, the region of the anastomosis could be clearly defined on the fused PET/CT images. PET/CT revealed a decreased concentration of FDG in the irradiated field 2 and 4 weeks after IORT. At 8 weeks, however, the distribution of the tracer in the irradiated pigs did not differ from that in the nonirradiated pigs. Homogeneous tracer uptake in all liver regions was observed in the nonirradiated animals. The CT images showed an increase in liver volume in all pigs and bile duct dilatation that increased over time in the irradiated pigs. CONCLUSION: The morphologic and functional changes due to IORT in liver tissue can be accurately monitored with dual-modality PET/CT. By enabling the integration of functional and morphologic data, PET/CT may have an important role in monitoring radiation treatment. PMID- 14764889 TI - Dual-modality PET/CT scanning with negative oral contrast agent to avoid artifacts: introduction and evaluation. AB - The authors qualitatively and quantitatively assessed a solution containing 0.2% locust bean gum (LBG) and 2.5% mannitol (mannitol-LBG) dissolved in water to provide a negative oral contrast material in dual-modality positron emission tomography (PET)/computed tomography (CT) scanning. PET/CT was performed in 60 patients with cancer after oral administration of barium, water, or mannitol-LBG. Qualitative and quantitative analyses were conducted to determine bowel distention and a potential influence of the contrast agents on the PET data. Intestinal distention with mannitol-LBG proved superior to that with water or barium. Findings at both quantitative and qualitative analysis revealed apparently increased tracer uptake in the small bowel with barium in comparison to that with mannitol-LBG or water. Mannitol-LBG may, therefore, be used as a negative oral contrast agent at PET/CT scanning because it provides excellent bowel distention while avoiding contrast material-induced PET artifacts. PMID- 14764890 TI - Heat-activated liposomal MR contrast agent: initial in vivo results in rabbit liver and kidney. AB - PURPOSE: To evaluate by using in vivo magnetic resonance (MR) imaging the functionality of a liposomal paramagnetic contrast agent with T1 relaxivity that rapidly and markedly increases at temperatures above the gel-to-liquid crystalline phase transition temperature (T(c)) of the liposome membrane. MATERIALS AND METHODS: Liposomal gadolinium diethylenetriaminepentaacetic acid bis(methylamide) was injected intravenously at a dose of 0.4 or 1.2 mL (containing 10 or 30 micromol of gadolinium, respectively) per kilogram of body weight shortly before the application of focused ultrasound in liver (seven rabbits) or kidney (three rabbits). VX2 tumors had been implanted in liver in four of the rabbits. Eighteen locations in liver (13 in normal tissue, five in tumor) and 12 locations in kidney were sonicated. MR thermometry was performed during sonications. Signal intensity enhancement was evaluated on T1-weighted images acquired after the tissue cooled, and enhanced zones were compared with isotherms at T(c) of the liposome membrane (approximately 57 degrees C) by using Bland-Altman analysis. In liver, enhanced zones also were compared with areas of histologically verified thermal damage. The threshold temperature of enhancement at T1-weighted imaging was verified by monitoring the signal intensity increase after 10 sonications at varied powers in two locations in normal liver tissue. RESULTS: Persistent enhancement was observed on T1-weighted images at all sonicated liver locations. In liver, enhanced zones on T1-weighted images were contiguous both with 57 degrees C isotherms (25 measurements; mean difference +/- SD, 0.4 mm +/- 1.2) and with histologically verified areas of necrosis (seven measurements; mean difference +/- SD, 0.1 mm +/- 0.9). The threshold temperature of enhancement at T1-weighted imaging in normal liver was 53 degrees -57 degrees C. In kidney, enhanced zones on T1-weighted images did not match the isotherms. CONCLUSION: The liposomal contrast agent was effective at in vivo MR thermometry in liver but not in kidney. PMID- 14764891 TI - Computer-aided detection output on 172 subtle findings on normal mammograms previously obtained in women with breast cancer detected at follow-up screening mammography. AB - PURPOSE: To evaluate, by using a computer-aided detection (CAD) program, the nonspecific findings on normal screening mammograms obtained in women in whom breast cancer was later detected at follow-up screening mammography. MATERIALS AND METHODS: Four hundred ninety-three mammogram pairs-an initial negative screening mammogram and a subsequently obtained screening mammogram showing cancer-were collected. The mean interval between examinations was 14.6 months. In 169 cases, in which 172 cancers were later depicted, findings on the initial mammogram were subtle enough that either none or only one or two of five blinded radiologists recommended screening recall. On the initial negative mammograms, of the 172 areas where cancer later developed, 137 (80%) had subtle nonspecific findings and were retrospectively judged as having a benign or normal appearance. The mammograms with these subtle findings were evaluated with a commercially available CAD program, and the numbers of CAD marks on these nonspecific findings were analyzed. RESULTS: Of the 172 cancers, 129 (75%) were invasive and 43 (25%) were ductal carcinoma in situ. The CAD program marked 72 (42%) of the 172 findings that subsequently developed into cancer: 24 (29%) of 82 findings recalled by none, 25 (49%) of 51 findings recalled by one, and 23 (59%) of 39 findings recalled by two of the five radiologists. Among the 137 areas with nonspecific normal or benign findings, 41 (30%) areas where cancer subsequently developed were marked by the CAD program. CONCLUSION: A subset of cancers have perceptible but nonspecific mammographic findings that may be marked by a CAD program, even when the findings do not warrant recall as judged at blinded and unblinded radiologist review. The authors believe failure to act on such nonspecific but CAD-marked findings prospectively does not constitute interpretation below a reasonable standard of care. PMID- 14764892 TI - LacZ-expressing olfactory ensheathing cells do not associate with myelinated axons after implantation into the compressed spinal cord. AB - Studies have shown that implanting olfactory ensheathing cells (OECs) may be a promising therapeutic strategy to promote functional recovery after spinal cord injury. Several fundamental questions remain, however, regarding their in vivo interactions in the damaged spinal cord. We have induced a clip compression injury at the T10 level of the spinal cord in adult rats. After a delay of 1 week, OECs isolated from embryonic day 18 rats were implanted into the cystic cavity that had formed at the site of injury. Before implantation, OECs were infected with a LacZ-expressing retrovirus. At 3 weeks after implantation, LacZ expressing OECs survived the implantation procedure and remained localized to the cystic cavity. At the electron microscopic level, the cystic cavity had clusters of LacZ-expressing OECs and numerous Schwann cells lacking LacZ expression. Although labeled OECs made no direct contact with axons, unlabeled Schwann cells were associated with either a single myelinated axon or multiple unmyelinated axons. Positively labeled OEC processes often enveloped multiple Schwann cell axon units. These observations suggest that the role of OECs as the primary mediators of the beneficial effects on axon growth, myelination, and functional recovery after spinal cord injury may require re-evaluation. PMID- 14764893 TI - Mice with cardiomyocyte-specific disruption of the endothelin-1 gene are resistant to hyperthyroid cardiac hypertrophy. AB - Endothelin 1 (ET-1), a potent vasoconstrictor peptide expressed by endothelium, is also produced in the heart in response to a variety of stresses. It induces hypertrophy in cultured cardiac myocytes but only at concentrations far greater than those found in plasma. We tested whether ET-1 generated by cardiac myocytes in vivo is a local signal for cardiac hypertrophy. To avoid the perinatal lethality seen in systemic ET-1-null mice, we used the Cre/loxP system to generate mice with cardiac myocyte-specific disruption of the ET-1 gene. We used the alpha-myosin heavy chain promoter to drive expression of Cre and were able to obtain 75% reduction in ET-1 mRNA in cardiac myocytes isolated from these mice at baseline and after stimulation, in vivo, for 24 h with tri-iodothyronine (T3). Necropsy measurements of cardiac mass indexed for body weight showed a 57% reduction in cardiac hypertrophy in response to 16 days of exogenous T3 in mice homozygous for the disrupted ET-1 allele compared to siblings with an intact ET-1 gene. Moreover, in vivo MRI showed only a 3% increase in left ventricular mass indexed for body weight in mice with the disrupted allele after 3 weeks of T3 treatment versus a 27% increase in mice with an intact ET-1 gene. A reduced hypertrophic response was confirmed by planimetry of cardiac myocytes. We conclude that ET-1, produced locally by cardiac myocytes, and acting in a paracrine/autocrine manner, is an important signal for myocardial hypertrophy that facilitates the response to thyroid hormone. PMID- 14764894 TI - Protection against electrophile and oxidant stress by induction of the phase 2 response: fate of cysteines of the Keap1 sensor modified by inducers. AB - Induction of a family of phase 2 genes encoding for proteins that protect against the damage of electrophiles and reactive oxygen intermediates is potentially a major strategy for reducing the risk of cancer and chronic degenerative diseases. Many phase 2 genes are regulated by upstream antioxidant response elements (ARE) that are targets of the leucine zipper transcription factor Nrf2. Under basal conditions, Nrf2 resides mainly in the cytoplasm bound to its cysteine-rich, Kelch domain-containing partner Keap1, which is itself anchored to the actin cytoskeleton and represses Nrf2 activity. Inducers disrupt the Keap1-Nrf2 complex by modifying two (C273 and C288) of the 25 cysteine residues of Keap1. The critical role of C273 and C288 was established by (i) their high reactivity when purified recombinant Keap1 was treated with dexamethasone mesylate and the dexamethasone-modified tryptic peptides were analyzed by mass spectrometry, and (ii) transfection of keap1 and nrf2 gene-deficient mouse embryonic fibroblasts with constructs expressing cysteine to alanine mutants of Keap1, and measurement of the ability of cotransfected Nrf2 to repress an ARE-luciferase reporter. Reaction of Keap1 with inducers results in formation of intermolecular disulfide bridges, probably between C273 of one Keap1 molecule and C288 of a second. Evidence for formation of such dimers was obtained by 2D PAGE of extracts of cells treated with inducers, and by the demonstration that whereas C273A and C288A mutants of Keap1 alone could not repress Nrf2 activation of the ARE luciferase reporter, an equal mixture of these mutant constructs restored repressor activity. PMID- 14764895 TI - The limits of protection by "memory" T cells in Ig-/- mice persistently infected with a gamma-herpesvirus. AB - Can CD4(+) and CD8(+) "memory" T cells that are generated and maintained in the context of low-level virus persistence protect, in the absence of antibody, against a repeat challenge with the same pathogen? Although immune T cells exert effective, long-term control of a persistent gamma-herpesvirus (gammaHV68) in Ig( /-) microMT mice, subsequent exposure to a high dose of the same virus leads to further low-level replication in the lung. This lytic phase in the respiratory tract is dealt with effectively by the recall of memory T cells induced by a gammaHV68 recombinant (M3LacZ) that does not express the viral M3 chemokine binding protein. At least for the CD8(+) response, greater numbers of memory T cells confer enhanced protection in the M3LacZ-immune mice. However, neither WT gammaHV68 nor the minimally persistent M3LacZ primes the T cell response to the extent that a WT gammaHV68 challenge fails to establish latency in the microMT mice. Memory CD4(+) and CD8(+) T cells thus act together to limit gammaHV68 infection but are unable to provide absolute protection against a high-dose, homologous challenge. PMID- 14764896 TI - The world of geography: visualizing a knowledge domain with cartographic means. AB - From an informed critique of existing methods to the development of original tools, cartographic engagement can provide a unique perspective on knowledge domain visualization. Along with a discussion of some principles underlying a cartographically informed visualization methodology, results of experiments involving several thousand conference abstracts will be sketched and their plausibility reflected on. PMID- 14764897 TI - The role of Shc and insulin-like growth factor 1 receptor in mediating the translocation of estrogen receptor alpha to the plasma membrane. AB - Our previous studies demonstrated that 17beta-estradiol (E2) rapidly induces the interaction of estrogen receptor alpha (ERalpha) with the adapter protein Shc, the translocation of ERalpha to the cell membrane, and the formation of dynamic membrane structures in MCF-7 breast cancer cells. The present study examined how E2 causes ERalpha to translocate to the region of the plasma membrane and focused on mechanisms whereby Shc and the insulin-like growth factor-1 receptor (IGF-1R) mediate this process. Shc physically interacts with IGF-1R in the plasma membrane, and E2 activates IGF-1R. We reasoned that ERalpha, when bound to Shc, would be directed to the region of the plasma membrane by the same processes, causing membrane translocation of Shc. We confirmed that E2 rapidly induced IGF 1R phosphorylation and demonstrated that E2 induced formation of a ternary protein complex among Shc, ERalpha, and IGF-1R. Knock down of Shc with a specific small inhibitory RNA decreased the association of ERalpha with IGF-1R by 87%, suggesting that Shc is a crucial molecule in the formation of this ternary complex. Confocal microscopy studies provided further confirmation of the functional roles of Shc and the IGF-1R in the translocation of ERalpha to the region of the membrane. Down-regulation of Shc, ERalpha, or IGF-1R with specific small inhibitory RNAs all blocked E2-induced mitogen-activated protein kinase phosphorylation. Together, our results demonstrate that Shc and IGF-1R serve as key elements in the translocation of ERalpha to the cell membrane and in the facilitation of ERalpha-mediated rapid E2 action. PMID- 14764898 TI - Scaffolding of Keap1 to the actin cytoskeleton controls the function of Nrf2 as key regulator of cytoprotective phase 2 genes. AB - Transcription factor Nrf2 regulates basal and inducible expression of phase 2 proteins that protect animal cells against the toxic effects of electrophiles and oxidants. Under basal conditions, Nrf2 is sequestered in the cytoplasm by Keap1, a multidomain, cysteinerich protein that is bound to the actin cytoskeleton. Keap1 acts both as a repressor of the Nrf2 transactivation and as a sensor of phase 2 inducers. Electrophiles and oxidants disrupt the Keap1-Nrf2 complex, resulting in nuclear accumulation of Nrf2, where it enhances the transcription of phase 2 genes via a common upstream regulatory element, the antioxidant response element. Reporter cotransfection-transactivation analyses with a series of Keap1 deletion mutants revealed that in the absence of the double glycine repeat domain Keap1 does not bind to Nrf2. In addition, deletion of either the intervening region or the C-terminal region also abolished the ability of Keap1 to sequester Nrf2, indicating that all of these domains contribute to the repressor activity of Keap1. Immunocytochemical and immunoprecipitation analyses demonstrated that Keap1 associates with actin filaments in the cytoplasm through its double glycine repeat domain. Importantly, disruption of the actin cytoskeleton promotes nuclear entry of an Nrf2 reporter protein. The actin cytoskeleton therefore provides scaffolding that is essential for the function of Keap1, which is the sensor for oxidative and electrophilic stress. PMID- 14764899 TI - Type I MADS-box genes have experienced faster birth-and-death evolution than type II MADS-box genes in angiosperms. AB - Plant MADS-box genes form a large gene family for transcription factors and are involved in various aspects of developmental processes, including flower development. They are known to be subject to birth-and-death evolution, but the detailed features of this mode of evolution remain unclear. To have a deeper insight into the evolutionary pattern of this gene family, we enumerated all available functional and nonfunctional (pseudogene) MADS-box genes from the Arabidopsis and rice genomes. Plant MADS-box genes can be classified into types I and II genes on the basis of phylogenetic analysis. Conducting extensive homology search and phylogenetic analysis, we found 64 presumed functional and 37 nonfunctional type I genes and 43 presumed functional and 4 nonfunctional type II genes in Arabidopsis. We also found 24 presumed functional and 6 nonfunctional type I genes and 47 presumed functional and 1 nonfunctional type II genes in rice. Our phylogenetic analysis indicated there were at least about four to eight type I genes and approximately 15-20 type II genes in the most recent common ancestor of Arabidopsis and rice. It has also been suggested that type I genes have experienced a higher rate of birth-and-death evolution than type II genes in angiosperms. Furthermore, the higher rate of birth-and-death evolution in type I genes appeared partly due to a higher frequency of segmental gene duplication and weaker purifying selection in type I than in type II genes. PMID- 14764900 TI - Expansion and contraction of the hepatitis B virus transcriptional template in infected chimpanzees. AB - We have previously shown that hepatitis B virus (HBV) replication is controlled by noncytolytic mechanisms that depend primarily on the effector functions of the CD8(+) T cell response, especially the production of IFN-gamma in the liver. The mechanisms that control the nuclear pool of viral covalently closed circular DNA (cccDNA) transcriptional template of HBV, which must be eliminated to eradicate infection, have been difficult to resolve. To examine those mechanisms, we quantitated intrahepatic HBV cccDNA levels in acutely infected chimpanzees whose virological, immunological, and pathological features were previously described. Our results demonstrate that the elimination kinetics of the cccDNA are more rapid than the elimination of HBV antigen-positive hepatocytes during the early phase of viral clearance, and they coincide with the influx of small numbers of IFN-gamma producing CD8(+) T cells into the liver. In contrast, terminal clearance of the cccDNA is associated with the peak of liver disease and hepatocellular turnover and with a surge of IFN-gamma producing CD8(+) T cells in the liver. Collectively, these results suggest that cccDNA clearance is a two step process mediated by the cellular immune response. The first step reduces the pool of cccDNA molecules noncytolytically, probably by eliminating their relaxed circular DNA precursors and perhaps by destabilizing them. The second step enhances this process by destroying infected hepatocytes and triggering their turnover. Surprisingly, despite this multipronged response, traces of cccDNA persist indefinitely in the liver, likely providing a continuous antigenic stimulus that confers lifelong immunity. PMID- 14764901 TI - Development of a specific system for targeting protein to metallophilic macrophages. AB - The cysteine-rich domain (CR) of the mannose receptor binds sulfated glycoprotein CR ligand (CRL) expressed by subpopulations of myeloid cells in secondary lymphoid organs (CRL(+) cells). In naive mice, these CRL(+) cells, metallophilic macrophages (M) in spleen and subcapsular sinus M in lymph nodes, are located strategically for antigen capture and are adjacent to B cell follicles, but their role in the immune response is unknown. We have exploited the lectin activity of CR to develop a highly specific system for targeting protein to CRL(+) M. We demonstrate that the sulfated carbohydrates recognized by CR are exposed to the extracellular milieu and mediate highly specific targeting of CR-containing proteins. This model will allow the dissection of the role of metallophilic M in an immune response in vivo. PMID- 14764902 TI - Two distinct signaling pathways participate in auxin-induced swelling of pea epidermal protoplasts. AB - Protoplast swelling was used to investigate auxin signaling in the growth limiting stem epidermis. The protoplasts of epidermal cells were isolated from elongating internodes of pea (Pisum sativum). These protoplasts swelled in response to auxin, providing the clearest evidence that the epidermis can directly perceive auxin. The swelling response to the natural auxin IAA showed a biphasic dose response curve but that to the synthetic auxin 1-naphthalene acetic acid (NAA) showed a simple bell-shaped dose response curve. The responses to IAA and NAA were further analyzed using antibodies raised against ABP1 (auxin-binding protein 1), and their dependency on extracellular ions was investigated. Two signaling pathways were resolved for IAA, an ABP1-dependent pathway and an ABP1 independent pathway that is much more sensitive to IAA than the former. The response by the ABP1 pathway was eliminated by anti-ABP1 antibodies, had a higher sensitivity to NAA, and did not depend on extracellular Ca(2+). In contrast, the response by the non-ABP1 pathway was not affected by anti-ABP1 antibodies, had no sensitivity to NAA, and depended on extracellular Ca(2+). The swelling by either pathway required extracellular K(+) and Cl(-). The auxin-induced growth of pea internode segments showed similar response patterns, including the occurrence of two peaks in the dose response curve for IAA and the difference in Ca(2+) requirements. It is suggested that two signaling pathways participate in auxin induced internode growth and that the non-ABP1 pathway is more likely to be involved in the control of growth by constitutive concentrations of endogenous auxin. PMID- 14764904 TI - Temporal dynamics of carbon partitioning and rhizodeposition in wheat. AB - The temporal dynamics of partitioning and rhizodeposition of recent photosynthate in wheat (Triticum aestivum) roots were quantified in situ in solution culture. After a 30-min pulse of (14)CO(2) to a single intact leaf, (14)C activities of individual carbon fluxes in the root, including exudation, respiration, and root content, were measured continuously over the next 20 h concurrently with (14)C efflux from the leaf. Immediately after the end of the (14)CO(2) pulse, (14)C activity was detected in the root, the hydroponic solution, and in root respiration. The rate of (14)C exudation from the root was maximal after 2 to 3 h, and declined to one-third of maximum after a further 5 h. Completion of the rapid phase of (14)C efflux from the leaf coincided with peak (14)C exudation rate. Thus, exudation flux is much more rapidly and dynamically coupled to current photosynthesis than has been appreciated. Careful cross-calibration of (14)C counting methods allowed a dynamic (14)C budget to be constructed for the root. Cumulative (14)C exudation after 20 h was around 3% of (14)C fixed in photosynthesis. Partitioning of photosynthate between shoot and root was manipulated by partial defoliation before applying the (14)CO(2) pulse to the remaining intact leaf. Although the rate of photosynthesis was largely unaffected by partial defoliation, the proportion of new photosynthate subsequently partitioned to and exuded from the root was substantially reduced. This clearly indicates that exudation depends more on the rate of carbon import into the root than on the rate of photosynthesis. PMID- 14764903 TI - The gar2 and rga alleles increase the growth of gibberellin-deficient pollen tubes in Arabidopsis. AB - Ectopic expression in Arabidopsis of a pea (Pisum sativum) cDNA (2ox2) encoding a gibberellin (GA) 2-oxidase (PsGA2ox2), involved in the deactivation of biologically active GAs, has been used to establish a role for GAs in promoting pollen tube growth. One line, 35S:2ox2/28c, when homozygous for the transgene, exhibits a novel small fruit phenotype. The 28c transgene reduces pollen tube growth, and this results in a reduced number of fertilized seeds that are only present at the end of the silique nearest the stigma. To confirm that the 28c pollen tube phenotype is due to sense expression of the 2ox2 mRNA, a "hairpin" RNA interface silencing construct, designed to silence 2ox2 expression, has been used to restore pollen tube growth and fruit development. The interaction between 28c and other mutants with increased GA response has also been examined to provide further evidence that GAs play an important role in pollen tube growth. Based on the ability of mutant alleles to suppress the 35S:2ox2/28c phenotype, we define new roles for the gar2-1 and rga alleles in GA signaling during pollen tube elongation in addition to their previously established roles in vegetative tissues. In contrast to the constitutive GA response observed in internodes and leaves lacking RGA and GAI, the rga-2 gai-d5 mutant combination is only a partial suppressor of the 28c phenotype. Because the dominant dwarfing gai-1 allele reduces GA response in vegetative tissues, its effect on plant fertility has been examined. Although gai-1 reduces seed set, this appears to reflect defects in reproductive development other than pollen tube function. Finally, we show that the genetic background (Landsberg erecta or Columbia) modifies the 28c phenotype and that this effect is not due to the ER/er difference between these two ecotypes. PMID- 14764905 TI - Cloning, characterization, and immunolocalization of a mycorrhiza-inducible 1 deoxy-d-xylulose 5-phosphate reductoisomerase in arbuscule-containing cells of maize. AB - Colonization of plant roots by symbiotic arbuscular mycorrhizal fungi frequently leads to the accumulation of several apocarotenoids. The corresponding carotenoid precursors originate from the plastidial 2-C-methyl-d-erythritol 4-phosphate pathway. We have cloned and characterized 1-deoxy-d-xylulose 5-phosphate reductoisomerase (DXR), catalyzing the first committed step of the pathway, from maize (Zea mays). Functional identification was accomplished by heterologous expression of sequences coding for the mature protein in Escherichia coli. DXR is up-regulated in maize roots during mycorrhization as shown at transcript and protein levels, but is also abundant in leaves and young seedlings. Inspection of sequenced genomes and expressed sequence tag (EST) databases argue for a single copy DXR gene. Immunolocalization studies in mycorrhizal roots using affinity purified antibodies revealed a DXR localization in plastids around the main symbiotic structures, the arbuscules. DXR protein accumulation is tightly correlated with arbuscule development. The highest level of DXR protein is reached around maturity and initial senescence of these structures. We further demonstrate the formation of a DXR-containing plastidial network around arbuscules, which is highly interconnected in the mature, functional state of the arbuscules. Our findings imply a functional role of a still unknown nature for the apocarotenoids or their respective carotenoid precursors in the arbuscular life cycle. PMID- 14764906 TI - PvUPS1, an allantoin transporter in nodulated roots of French bean. AB - Nodulated legumes receive their nitrogen via nitrogen-fixing rhizobia, which exist in a symbiotic relationship with the root system. In tropical legumes like French bean (Phaseolus vulgaris) or soybean (Glycine max), most of the fixed nitrogen is used for synthesis of the ureides allantoin and allantoic acid, the major long-distance transport forms of organic nitrogen in these species. The purpose of this investigation was to identify a ureide transporter that would allow us to further characterize the mechanisms regulating ureide partitioning in legume roots. A putative allantoin transporter (PvUPS1) was isolated from nodulated roots of French bean and was functionally characterized in an allantoin transport-deficient yeast mutant showing that PvUPS1 transports allantoin but also binds its precursors xanthine and uric acid. In beans, PvUPS1 was expressed throughout the plant body, with strongest expression in nodulated roots, source leaves, pods, and seed coats. In roots, PvUPS1 expression was dependent on the status of nodulation, with highest expression in nodules and roots of nodulated plants compared with non-nodulated roots supplied with ammonium nitrate or allantoin. In situ RNA hybridization localized PvUPS1 to the nodule endodermis and the endodermis and phloem of the nodule vasculature. These results strengthen our prediction that in bean nodules, PvUPS1 is involved in delivery of allantoin to the vascular bundle and loading into the nodule phloem. PMID- 14764907 TI - Coordinated regulation of genes for secretion in tobacco at late developmental stages: association with resistance against oomycetes. AB - Besides the systemic acquired resistance (SAR) induced in response to microbial stimulation, host plants may also acquire resistance to pathogens in response to endogenous stimuli associated with their own development. In tobacco (Nicotiana tabacum), the vegetative-to-flowering transition comes along with a susceptibility-to-resistance transition to the causal agent of black shank disease, the oomycete Phytophthora parasitica. This resistance affects infection effectiveness and hyphal expansion and is associated with extracellular accumulation of a cytotoxic activity that provokes in vitro cell death of P. parasitica zoospores. As a strategy to determine the extracellular events important for restriction of pathogen growth, we screened the tobacco genome for genes encoding secreted or membrane-bound proteins expressed in leaves of flowering plants. Using a signal sequence trap approach in yeast (Saccharomyces cerevisiae), 298 clones were selected that appear to encode for apoplastic, cell wall, or membrane-bound proteins involved in stress response, in plant defense, or in cell wall modifications. Microarray and northern-blot analyses revealed that, at late developmental stages, leaves were characterized by the coordinate up-regulation of genes involved in SAR and in peroxidative cross-linking of structural proteins to cell wall. This suggests the potential involvement of these genes in extracellular events that govern the expression of developmental resistance. The analysis of the influence of salicylic acid on mRNA accumulation also indicates a more complex network for regulation of gene expression at a later stage of tobacco development than during SAR. Further characterization of these genes will permit the formulation of hypotheses to explain resistance and to establish the connection with development. PMID- 14764909 TI - Dual-function ion channel/protein kinases: novel components of vertebrate magnesium regulatory mechanisms. AB - Although magnesium is the dominant divalent intracellular cation and is required for the function of diverse types of enzymes that participate in virtually every cellular process, the molecular mechanisms that regulate its homeostasis are poorly understood. Electrophysiologic and biochemical investigations of a novel dual-function ion channel/kinase protein have recently converged with the identification of the gene locus for an autosomal recessive form of inherited hypomagnesemia to provide new insight into vertebrate magnesium regulatory mechanisms. PMID- 14764908 TI - Lipoic acid-dependent oxidative catabolism of alpha-keto acids in mitochondria provides evidence for branched-chain amino acid catabolism in Arabidopsis. AB - Lipoic acid-dependent pathways of alpha-keto acid oxidation by mitochondria were investigated in pea (Pisum sativum), rice (Oryza sativa), and Arabidopsis. Proteins containing covalently bound lipoic acid were identified on isoelectric focusing/sodium dodecyl sulfate-polyacrylamide gel electrophoresis separations of mitochondrial proteins by the use of antibodies raised to this cofactor. All these proteins were identified by tandem mass spectrometry. Lipoic acid containing acyltransferases from pyruvate dehydrogenase complex and alpha ketoglutarate dehydrogenase complex were identified from all three species. In addition, acyltransferases from the branched-chain dehydrogenase complex were identified in both Arabidopsis and rice mitochondria. The substrate-dependent reduction of NAD(+) was analyzed by spectrophotometry using specific alpha-keto acids. Pyruvate- and alpha-ketoglutarate-dependent reactions were measured in all three species. Activity of the branched-chain dehydrogenase complex was only measurable in Arabidopsis mitochondria using substrates that represented the alpha-keto acids derived by deamination of branched-chain amino acids (Val [valine], leucine, and isoleucine). The rate of branched-chain amino acid- and alpha-keto acid-dependent oxygen consumption by intact Arabidopsis mitochondria was highest with Val and the Val-derived alpha-keto acid, alpha-ketoisovaleric acid. Sequencing of peptides derived from trypsination of Arabidopsis mitochondrial proteins revealed the presence of many of the enzymes required for the oxidation of all three branched-chain amino acids. The potential role of branched-chain amino acid catabolism as an oxidative phosphorylation energy source or as a detoxification pathway during plant stress is discussed. PMID- 14764910 TI - Maternally administered dexamethasone at 0.7 of gestation suppresses maternal and fetal pituitary and adrenal responses to hypoxemia in sheep. AB - Women who are at risk of preterm delivery are treated with antenatal steroids to facilitate fetal lung maturation. During this period, there is a potential for fetal or maternal hypoxemia to occur. Fetal responses to hypoxemia in sheep are well documented. However, less is known regarding maternal responses to hypoxemia. Therefore, we determined the effects of dexamethasone (DM) on maternal and fetal responses to hypoxemia in sheep. Ewes received four i.m. injections of DM or saline at 12-h intervals beginning at 103 d of gestation. Samples for ACTH, cortisol, and glucose were collected at 0900 h. At 105 d of gestation, hypoxemia was induced for 1 h by maternal nitrogen gas inhalation. Samples for ACTH, cortisol, and glucose were collected at 15-min intervals before, during, and after the hypoxemia challenge. Fluorescent microspheres were administered to the mother and the fetus before and during hypoxemia to measure organ perfusion. DM suppressed basal fetal and maternal cortisol and ACTH concentrations but increased glucose levels. DM also increased fetal but not maternal blood pressure. In control subjects, hypoxemia elevated fetal and maternal cortisol and ACTH concentrations. These responses were obliterated by DM. Hypoxemia increased blood pressure in DM-exposed fetuses but not in control subjects. In addition, hypoxemia decreased fetal adrenal vascular resistance in saline but not DM fetuses or ewes from either treatment group. In summary, maternal administration of a low dose of DM at 0.7 of gestation suppresses maternal and fetal adrenal function and changes fetal responses to hypoxemic stress to resemble those observed later in gestation. PMID- 14764911 TI - Effects of enterally administering granulocyte colony-stimulating factor to suckling mice. AB - Gastrointestinal (GI) tract development is influenced by multiple growth factors, some of which are delivered directly to the GI lumen, as they are swallowed constituents of amniotic fluid, colostrum, and milk. Granulocyte colony stimulating factor (G-CSF), traditionally known as a granulocytopoietic growth factor, is an example of one such factor. However, it is not clear whether the large amounts of G-CSF that are normally swallowed by the fetus and neonate have systemic effects on circulating neutrophils or local effects in the developing intestine. To assess this, we administered either active or heat-denatured (control) recombinant human G-CSF to 5- to 7-d-old C57BL/6 x 129SvJ mice. Pups received either a low dose (3 ng) that was calculated to approximate the amount of G-CSF swallowed in utero from amniotic fluid or an isovolemic high dose 100 times larger (300 ng). Oral dosing was performed daily for either 3 or 7 d, after which pups were killed and measurements were made on the blood and the GI tract. Absolute blood neutrophil counts and immature to total neutrophil ratios did not differ from controls in any of the test groups. However, intestinal villus area, perimeter, length, crypt depth, and proliferating cell nuclear antigen index increased significantly among those that were treated with active G-CSF. Thus, in suckling mice, enterally administered G-CSF had no effect on the concentration of circulating neutrophils but had trophic effects on the intestine. We speculate that the G-CSF present in amniotic fluid, colostrum, and milk acts as a topical intestinal growth factor and has little or no granulocytopoietic action. PMID- 14764912 TI - Beta-carotene prevents bile acid-induced cytotoxicity in the rat hepatocyte: Evidence for an antioxidant and anti-apoptotic role of beta-carotene in vitro. AB - Hydrophobic bile acids are implicated in the pathogenesis of cholestatic liver disorders through mechanisms involving oxidative stress and mitochondrial dysfunction. Antioxidants ameliorate bile acid-induced cytotoxicity in rat hepatocyte suspensions. The purpose of the current study was to evaluate the potential protective role of beta-carotene (betaC), a putative fat-soluble antioxidant that is reduced in patients with cholestasis, against bile acid induced hepatotoxicity. In freshly isolated rat hepatocyte suspensions that were exposed to the toxic hydrophobic bile acid glycochenodeoxycholic acid (100 or 500 microM), betaC (100 microM) decreased generation of reactive oxygen species by >50%, similar to the inhibition afforded by alpha-tocopherol. Commensurate with this antioxidant effect, 100 microM betaC also protected hepatocytes against both glycochenodeoxycholic acid-induced cellular necrosis and apoptosis, which was associated with reduction in caspase 3 activation, inhibition of mitochondrial cytochrome c release in rat hepatocytes, and prevention of the mitochondrial permeability transition in both liver mitochondria and rat hepatocytes. A lower concentration of betaC (50 microM) produced similar antioxidant and anti apoptotic protection but with less inhibition against cell necrosis, suggesting that the higher concentration of betaC may have conferred additional cytoprotection not directly related to its antioxidant function. These results demonstrate that the antioxidant effects of betaC may provide hepatoprotection against cholestatic liver injury by preventing bile acid-induced oxidative stress and mitochondrial perturbations. PMID- 14764914 TI - Caudate and hippocampal volumes, intelligence, and motor impairment in 7-year-old children who were born preterm. AB - Children who survive very preterm birth without major disability have a high prevalence of learning difficulty, attention deficit, and minor motor impairment (MMI). To determine whether these difficulties are associated with structural brain abnormalities, we studied 105 preterm children (<32 wk) at 7 y with tests of IQ and MMI (Movement ABC) and detailed magnetic resonance brain scans. Scans were assessed qualitatively for visible cerebral lesions. Volume measurements of the caudate nuclei and hippocampal formations were made. Total brain volume (TBV) was estimated from the head circumference. Qualitative assessment of the scans showed evidence of cerebral lesions in 20 (19%), which were associated with lower IQ and more frequent MMI. IQ correlated with right and left caudate volume (Spearman's rho 0.304 and 0.349; p < 0.01). This association persisted (except for verbal IQ) when caudate volume was expressed as a percentage of estimated TBV to allow for overall brain size. No significant correlations with hippocampal volumes were observed. These differences persisted when only scans from children without visible lesions on scan were considered. MMI was significantly associated only with TBV and was more common in children with evidence of thinning of the posterior corpus callosum, although most children with MMI have a normal corpus callosum. Lower IQs in children who were born preterm are related to poorer development of the caudate relative to the rest of the brain, independent of other lesions. These findings suggest abnormal brain development after perinatal injury or postnatal nutritional deficits is responsible for cognitive deficits in preterm children. PMID- 14764913 TI - A new mitochondrial DNA mutation in ND3 gene causing severe Leigh syndrome with early lethality. AB - We describe a new mitochondrial DNA mutation in a male infant who presented clinical and magnetic resonance imaging features of Leigh syndrome and died at the age of 9 mo. The patient's development was reportedly normal in the first months of life. At the age of 5 mo, he presented severe generalized hypotonia, nystagmus, and absent eye contact. Laboratory examination showed increased lactate and pyruvate in both serum and cerebrospinal fluid. Brain magnetic resonance imaging revealed multiple necrotic lesions in the basal ganglia, brain stem, and thalamus. Muscle histopathology was unremarkable, whereas respiratory chain enzyme analysis revealed a severe complex I deficiency. The patient died after an acidotic coma at age 9 mo. Sequence analysis of the entire mtDNA disclosed a new T10158C mutation with variable tissue heteroplasm (muscle: 83%; blood: 48%). The mutation was undetectable in the blood of his unaffected mother. The transition changes a serine residue into a proline, in a highly conserved region of the NADH dehydrogenase subunit 3 (ND3). This is the first description of a mitochondrial ND3 gene in Leigh syndrome with early lethality. PMID- 14764915 TI - Tissue expression of nephrin in human and pig. AB - Nephrin is a major component of the glomerular filtration barrier. Mutations in the nephrin gene (NPHS1) are responsible for congenital nephrotic syndrome of the Finnish type (NPHS1). Nephrin was at first thought to be podocyte specific, but recent studies have suggested that nephrin is also expressed in nonrenal tissues such as pancreas and CNS. We studied the expression of nephrin in human and porcine tissues at different stages of development and correlated these findings to clinical characteristics of NPHS1 children. Immunofluorescence staining and Western blotting were used to detect nephrin protein in frozen tissue samples. Polyclonal antibodies against the intracellular part of nephrin were used in these analyses. In situ hybridization was used to detect nephrin mRNA in specimens from normal human subjects and patients with NPHS1. Nephrin protein was not detected in nonrenal tissues obtained from human and porcine fetuses, newborns, and infants. Likewise, nephrin mRNA expression was not observed outside kidney glomerulus in normal or NPHS1 children. The phenotype analysis of NPHS1 children with severe nephrin gene mutations supported the findings in the tissue expression studies and revealed no impairment of the neurologic, testicular, or pancreatic function in a great majority of the patients. The studies suggest that nephrin has no major clinical significance outside the kidney. PMID- 14764916 TI - Ventilatory responses to hypercapnia and hypoxia in conscious cystic fibrosis knockout mice Cftr-/-. AB - This study was designed to examine the ventilatory performance and the lung histopathology of cystic fibrosis knockout mice (Cftr-/-) compared with heterozygous (Cftr+/-) or wild-type (Cftr+/+) littermates. Ventilation was recorded in conscious animals using whole-body plethysmography. Tidal volume (VT), respiratory frequency (f), and minute ventilation (VE) were measured during air breathing and in response to various levels of hypercapnia (2, 4, 6, or 8% CO2) or hypoxia (14, 12, 10, or 8% O2). The results for Cftr+/- and Cftr+/+ were pooled into one control group because they did not differ. In air and in response to hypercapnia, VE, VT, and f were similar in Cftr-/- mice and in controls. During graded hypoxia, VE was decreased in Cftr-/- mice at 10 and 8% O2 because of a lower f. Histology showed neither inflammation nor obstruction of airways in Cftr-/- mice. Morphometric analysis showed alveolar dilation as a result of either distension or impaired development. In conclusion, cystic fibrosis knockout mice have normal baseline breathing and ventilatory response to hypercapnia but a decreased ventilatory response to severe hypoxia. This latter result associated with the morphometric analysis suggests that Cftr-/- mice may exhibit immaturity of the respiratory system. PMID- 14764917 TI - Report on the 2nd World Congress on Fetal Origins of Adult Disease, Brighton, U.K., June 7-10, 2003. AB - In 1989, reports suggested that the fetal environment, as reflected in birth size, was related to the risk of noncommunicable diseases in adult life. This association was first described for coronary heart disease but rapidly extended to include type 2 diabetes, osteoporosis, and metabolic and endocrine homeostasis. This led to the development of the fetal origins of adult disease paradigm, which resulted in a refocusing of research effort over the next 10 y to consider the lifelong consequences of perinatal influences on chronic diseases. Previously, perinatal influences had largely been seen in terms of teratogenic effects or acute birth injury rather than whether trajectories and responses made during early development had lifelong consequences. Indeed, in developmental biology, it is widely recognized that adaptive plastic responses during early development often have consequences for function in later adulthood. Although the relative importance of this newly recognized set of phenomena to the burden of human disease has been controversial, the research precipitated by those early observations has confirmed their robustness and started to provide a mechanistic basis to this biology. Two world congresses have been held to review progress in this research. Both have been characterized by a unique multidisciplinary attendance ranging from molecular, experimental, and developmental biologists to epidemiologists and health economists. PMID- 14764918 TI - Endothelial function and mechanical properties of the common carotid artery in children on parenteral nutrition. AB - Intravenous administration of nutrition mixtures induces endothelial damage and arterial wall remodeling in animal models. To study endothelial function and common carotid artery mechanical properties in children receiving parenteral nutrition, we used noninvasive ultrasonic measurements in 18 children on parenteral nutrition and 18 controls. No difference appeared in the geometry of the common carotid artery (intima media thickness, systolic and diastolic diameters) between the patients on parenteral nutrition and the controls. The incremental elastic modulus was significantly higher in the patients on parenteral nutrition (1.8 +/- 0.4 versus 1.4 +/- 0.5 4 mm Hg x 10(3), p < 0.05) reflecting alteration of the elastic properties of the arterial wall independent of the vessel geometry. The flow-mediated dilatation of the brachial artery was significantly lower in the patients on parenteral nutrition (6 +/- 3 versus 8 +/- 3%, p < 0.05), whereas the dilatation after glyceryl trinitrate administration was similar (22 +/- 9 versus 25 +/- 9%). Children on parenteral nutrition exhibit endothelial dysfunction and altered stiffness of the common carotid artery. The noninvasive methods used in this study may prove useful for objectively determining the effects of various preventive methods. PMID- 14764919 TI - Lipid peroxidation, caspase-3 immunoreactivity, and pyknosis in late-gestation fetal sheep brain after umbilical cord occlusion. AB - Umbilical cord occlusion (UCO), a known risk factor for perinatal brain damage, causes severe fetal asphyxia leading to oxidative stress, lipid peroxidation, and cell death. We have determined the effects of two 10-min UCO on the distribution of the lipid peroxidation marker 4-hydroxynonenal (4-HNE) and the activated form of the apoptosis marker caspase-3 in the brains of late-gestation fetal sheep. UCO caused asphyxia, hypertension, and bradycardia, but these parameters normalized 2 h after the occlusion. At postmortem, 48 h after the second UCO there were significantly higher numbers of 4-HNE-positive cells in all layers of the hippocampus and cerebellum, the parietal cortex, substantia nigra, caudate nucleus, putamen, and thalamus compared with control brains. 4-HNE immunoreactivity was also found in white matter tracts of the subcallosal bundle, external medullary lamina, reticular thalamic nucleus, and cerebellar fiber tracts only in UCO brains. Double-labeling identified these cells as predominantly neurons and astrocytes, with oligodendrocytes showing lower levels of 4-HNE immunoreactivity. After UCO, the number of caspase-3-immunopositive cells was increased significantly in the hippocampal CA1, molecular layer and dentate gyrus, ventrolateral thalamic nucleus, substantia nigra, putamen, and cerebellar granular and molecular layers compared with controls. Double-labeling revealed caspase-3 immunoreactivity was mainly in neurons, and to lesser extent in astrocytes and oligodendrocytes. Pyknotic cell numbers were significantly increased in hippocampal CA1 and CA3, parietal cortex, caudate nucleus, putamen, and cerebellar Purkinje cells after UCO. These data indicate that brief asphyxia induces widespread lipid peroxidation involving all cell types of the fetal brain and apoptosis in both neurons and glia. PMID- 14764920 TI - Enhanced iNOS expression in leukocytes and circulating endothelial cells is associated with the progression of coronary artery lesions in acute Kawasaki disease. AB - Nitric oxide (NO) serves many vasoprotective roles, but the massive release of NO causes arterial wall degeneration. We investigated whether enhanced nitric oxide synthase (iNOS) expression in peripheral blood leukocytes and circulating endothelial cells mirrors the progression of coronary arterial lesions in 55 children with acute Kawasaki disease (KD), including 24 with and 31 without coronary artery lesions (CAL). Patients were treated with i.v. gamma-globulin at the time of diagnosis and blood samples were collected before and after treatment. The cellular origin of NO synthesis was determined by flow cytometric analysis of iNOS expression in peripheral blood, and by immunohistochemical analysis of circulating endothelial cells and coronary arteries. iNOS expression in neutrophils peaked at the time of diagnosis, but did not peak in monocytes until 2 wk post onset of disease. Levels were significantly higher in both cell types in patients with CAL (p = 0.001 and p = 0.035, respectively). In addition, the number of circulating endothelial cells and levels of iNOS expression were higher in patients with CAL (p = 0.011 and p = 0.012, respectively). Immunohistochemical analysis of the coronary arteries from three patients with acute KD revealed iNOS immunoreactivity in endothelial cells, as well as infiltrating monocytes/macrophages in the aneurysms. We conclude that the expression of iNOS in peripheral blood leukocytes, as well as circulating endothelial cells, correlates with the severity of coronary arterial wall injury and the progression of CAL in patients with acute KD. PMID- 14764921 TI - Intestinal epithelial apoptosis initiates gross bowel necrosis in an experimental rat model of neonatal necrotizing enterocolitis. AB - The histopathology of necrotizing enterocolitis (NEC) is characterized by destruction of the mucosal layer in initial stages and by transmural necrosis of the intestinal wall in advanced stages of the disease. To test the hypothesis that enhanced epithelial apoptosis is an initial event underlying the gross histologic changes, we analyzed epithelial apoptosis and tissue morphology in an animal model of NEC and evaluated the effect of caspase inhibition on the incidence of experimental NEC in this model. Apoptosis was analyzed with terminal deoxynucleotidyltransferase-mediated dUTP-FITC nick end labeling (TUNEL) staining in intestinal sections and by measuring caspase 3 activity from intestinal lysates of neonatal rats subjected to formula feeding and cold/asphyxia stress (FFCAS) and from mother-fed (MF) controls. Morphologic evaluation was based on hematoxylin and eosin staining of intestinal sections. FFCAS resulted in histologic changes consistent with NEC, which were absent from MF animals. FFCAS was also associated with a significantly increased rate of nuclear DNA fragmentation in the small intestinal epithelium compared with MF. Elevated tissue caspase 3 activity confirmed the presence of apoptosis in samples with increased DNA fragmentation. Analysis of the coincidence of morphologic damage and apoptosis in corresponding tissue sections indicated that apoptosis precedes gross morphologic changes in this model. Furthermore, supplementation of formula with 8 boc-aspartyl(OMe)-fluoromethylketone, a pan-caspase inhibitor, significantly reduced the incidences of apoptosis and experimental NEC. These findings indicate that in neonatal rats FFCAS induces epithelial apoptosis that serves as an underlying cause for subsequent gross tissue necrosis. PMID- 14764922 TI - Developmental regulation of the rabbit blood-brain barrier LAT1 large neutral amino acid transporter mRNA and protein. AB - The expression of the blood-brain barrier (BBB) LAT1 large neutral amino acid transporter mRNA and protein was investigated in development in rabbits. The BBB LAT1 mRNA was down-regulated with postnatal development. However, the BBB immunoreactive LAT1 protein was unchanged in postnatal development, despite an up regulation of the BBB GLUT1 glucose transporter protein during this period. The dissociation between LAT1 protein and mRNA levels in development is consistent with posttranscriptional regulation of BBB LAT1 gene expression. PMID- 14764923 TI - Circulating angiogenic factors and the risk of preeclampsia. AB - BACKGROUND: The cause of preeclampsia remains unclear. Limited data suggest that excess circulating soluble fms-like tyrosine kinase 1 (sFlt-1), which binds placental growth factor (PlGF) and vascular endothelial growth factor (VEGF), may have a pathogenic role. METHODS: We performed a nested case-control study within the Calcium for Preeclampsia Prevention trial, which involved healthy nulliparous women. Each woman with preeclampsia was matched to one normotensive control. A total of 120 pairs of women were randomly chosen. Serum concentrations of angiogenic factors (total sFlt-1, free PlGF, and free VEGF) were measured throughout pregnancy; there were a total of 655 serum specimens. The data were analyzed cross-sectionally within intervals of gestational age and according to the time before the onset of preeclampsia. RESULTS: During the last two months of pregnancy in the normotensive controls, the level of sFlt-1 increased and the level of PlGF decreased. These changes occurred earlier and were more pronounced in the women in whom preeclampsia later developed. The sFlt-1 level increased beginning approximately five weeks before the onset of preeclampsia. At the onset of clinical disease, the mean serum level in the women with preeclampsia was 4382 pg per milliliter, as compared with 1643 pg per milliliter in controls with fetuses of similar gestational age (P<0.001). The PlGF levels were significantly lower in the women who later had preeclampsia than in the controls beginning at 13 to 16 weeks of gestation (mean, 90 pg per milliliter vs. 142 pg per milliliter, P=0.01), with the greatest difference occurring during the weeks before the onset of preeclampsia, coincident with the increase in the sFlt-1 level. Alterations in the levels of sFlt-1 and free PlGF were greater in women with an earlier onset of preeclampsia and in women in whom preeclampsia was associated with a small-for-gestational-age infant. CONCLUSIONS: Increased levels of sFlt-1 and reduced levels of PlGF predict the subsequent development of preeclampsia. PMID- 14764924 TI - Preeclampsia -- searching for the cause. PMID- 14764925 TI - Gene structure conservation aids similarity based gene prediction. AB - One of the primary tasks in deciphering the functional contents of a newly sequenced genome is the identification of its protein coding genes. Existing computational methods for gene prediction include ab initio methods which use the DNA sequence itself as the only source of information, comparative methods using multiple genomic sequences, and similarity based methods which employ the cDNA or protein sequences of related genes to aid the gene prediction. We present here an algorithm implemented in a computer program called Projector which combines comparative and similarity approaches. Projector employs similarity information at the genomic DNA level by directly using known genes annotated on one DNA sequence to predict the corresponding related genes on another DNA sequence. It therefore makes explicit use of the conservation of the exon-intron structure between two related genes in addition to the similarity of their encoded amino acid sequences. We evaluate the performance of Projector by comparing it with the program Genewise on a test set of 491 pairs of independently confirmed mouse and human genes. It is more accurate than Genewise for genes whose proteins are <80% identical, and is suitable for use in a combined gene prediction system where other methods identify well conserved and non-conserved genes, and pseudogenes. PMID- 14764926 TI - Two mutations in the tetracycline repressor change the inducer anhydrotetracycline to a corepressor. AB - We report for the first time the in vitro characterization of a reverse tetracycline repressor (revTetR). The dimeric wild-type repressor (TetR) binds to tet operator tetO in the absence of the inducer anhydrotetracycline (atc) to confer tight repression. We have isolated the revTetR G96E L205S mutant, which, contrary to TetR, binds tetO only in the presence of atc. This reverse acting mutant was overproduced and purified. Effector and DNA binding properties were analyzed by EMSA and quantified by fluorescence titration and surface plasmon resonance. The association constant K(A) of revTetR for binding of [atcMg](+) is approximately 10(8) M(-1), four orders of magnitude lower than that of TetR. The affinity of TetR for tetO is 5.6 +/- 2 x 10(9) M(-1) and that for revTetR in the presence of atc is 1 +/- 0.2 x 10(8) M(-1). Both induced forms, the atc-bound TetR and the free revTetR, have the same low affinity of 4 +/- 1 x 10(5) M(-1) for DNA. Therefore, atc does not act as a dimerization agent for revTetR. We discuss the structural differences between TetR and revTetR potentially underlying this reversal of activity. PMID- 14764929 TI - Ten-year risk of first recurrent stroke and disability after first-ever stroke in the Perth Community Stroke Study. AB - BACKGROUND AND PURPOSE: Limited information exists on the long-term prognosis after first-ever stroke. We aimed to determine the absolute frequency of first recurrent stroke and disability and the relative frequency of recurrent stroke over 10 years after first-ever stroke in Perth, Western Australia. METHODS: For a 12-month period beginning February 1989, all individuals with suspected acute stroke or transient ischemic attack who lived in a geographically defined and representative region of Perth were registered prospectively. Patients with a definite first-ever stroke were followed up 10 years after the index event. RESULTS: Over 10 years of follow-up, the cumulative risk of a first recurrent stroke was 43% (95% confidence interval [CI], 34 to 51). After the first year after first-ever stroke, the average annual risk of recurrent stroke was approximately 4%. Case fatality at 30 days after first recurrent stroke was 41%, which was significantly greater than the case fatality at 30 days after first ever stroke (22%) (P=0.003). For 30-day survivors of first-ever stroke, the 10 year cumulative risk of death or new institutionalization was 79% (95% CI, 73 to 85) and of death or new disability was 87% (95% CI, 81 to 92). CONCLUSIONS: Over 10 years of follow-up, the risk of first recurrent stroke is 6 times greater than the risk of first-ever stroke in the general population of the same age and sex, almost one half of survivors remain disabled, and one seventh require institutional care. Effective strategies for prevention of stroke need to be implemented early, monitored frequently, and maintained long term after first ever stroke. PMID- 14764930 TI - Influence of gender on K+-induced cerebral vasodilatation. AB - BACKGROUND AND PURPOSE: It is not known whether cerebral vasoprotective mechanisms in females include increased function of arterial K+ channels. We hypothesized that vasodilator responses mediated by activation of inwardly rectifying K+ (K(IR)) channels are greater in cerebral arteries of female versus male rats and that this is due to the effects of estrogen. METHODS: Changes in basilar artery diameter were measured with a cranial window preparation in anesthetized Sprague-Dawley rats. RESULTS: K+ (5 and 10 mmol/L) caused greater vasodilatation in females (percent maximum, 21+/-3% and 58+/-7%, respectively) versus males (11+/-1% and 37+/-4%, respectively; P<0.05). In contrast, vasodilator responses to aprikalim (1 and 3 micromol/L) or acetylcholine (ACh, 1 and 10 micromol/L) did not differ between the genders. The selective K(IR) channel inhibitor barium ion (30 micromol/L) decreased basilar artery diameter in males but not females (-7+/-1% versus -2+/-1%, P<0.05) and selectively inhibited K+-induced vasodilatation by approximately 50% in both groups. Ovariectomy of female rats resulted in smaller vasodilator effects of K+, and chronic treatment of these rats with 17beta-estradiol (0.01 mg/kg per day for 7 days) normalized K+ induced vasodilatation. Furthermore, the selective M2 muscarinic ACh receptor antagonist methoctramine (1 micromol/L) increased responses to K+ in males to levels equivalent to responses in females but had no effect on responses to K+ in females. CONCLUSIONS: K+ is a more powerful vasodilator in the female versus male cerebral circulation. This difference is estrogen dependent and could be due to a lack of M2 muscarinic ACh receptor-induced inhibition of K(IR) channel activation by K+ in female cerebral arteries. PMID- 14764931 TI - Risk of intracerebral hemorrhage in patients with arterial versus cardiac origin of cerebral ischemia on aspirin or placebo: analysis of individual patient data from 9 trials. AB - BACKGROUND AND PURPOSE: Patients who are anticoagulated after cerebral ischemia have a 19-fold-higher risk of intracerebral hemorrhage (ICH) if they had an arterial rather than a cardiac source. To determine whether this excess risk of ICH was due to the underlying disease (cerebral ischemia of arterial versus cardiac origin) or whether it depended on the antithrombotic regimen, we studied the risk of ICH in arterial versus cardiac origin of cerebral ischemia in patients who received aspirin or no antithrombotic drugs. METHODS: Individual patient data of patients who received aspirin or placebo after cerebral ischemia were obtained from 9 clinical trials. Presence of atrial fibrillation was considered evidence of a cardiac source. Otherwise, events were considered of arterial origin. Cox proportional-hazards modeling was used for univariate and multivariate analyses. RESULTS: Fifty-four ICHs occurred in 16 625 patient-years in the aspirin-treated patients, and 7 ICHs occurred in 4317 patient-years in those on placebo. After multivariate adjustment for age, sex, current smoking, history of hypertension and diabetes, and aspirin dose (aspirin-treated patients only), the hazard ratio for ICH in patients with an arterial versus a cardiac source was 0.74 (95% confidence interval, 0.30 to 1.82) for aspirin-treated patients and 4.34 (95% confidence interval, 0.35 to 54) for placebo-randomized patients. CONCLUSIONS: Our findings do not confirm the previous finding of an excess risk of ICH in patients with cerebral ischemia of arterial origin. Therefore, it seems that having cerebral ischemia of arterial origin by itself is not associated with an increased risk of ICH, but only in combination with high intensity anticoagulation. PMID- 14764932 TI - Concordance rate differences of 3 noninvasive imaging techniques to measure carotid stenosis in clinical routine practice: results of the CARMEDAS multicenter study. AB - BACKGROUND AND PURPOSE: To replace digital subtraction angiography (DSA) in carotid stenosis evaluation, noninvasive imaging techniques have to reach a high concordance rate. Our purpose is to compare the concordance rates of contrast enhanced MR angiography (CEMRA) and CT angiography (CTA) with Doppler ultrasound (DUS) in clinical routine practice. METHODS: We evaluated prospectively with DUS, CEMRA, and CTA 150 patients suspected of carotid stenosis. The overall concordance rates of the 3 techniques were calculated for symptomatic stenosis > or =50% and > or =70%, for asymptomatic stenosis > or =60%, and for occlusion. For the carotid arteries treated by surgery (n=97), the results of each method and combined techniques were recorded, and misclassification rates were evaluated from surgical reports. RESULTS: The overall concordance rates of DUS-CEMRA, DUS CTA, and CEMRA-CTA were not statistically different. However, the concordance rate of DUS-CEMRA (92.53%) was significantly higher than that for DUS-CTA (79.10%) in the surgical asymptomatic stenosis group (P=0.0258). CTA considered alone would misclassify the stenosis in a significant number of cases (11 of 64) in the surgical asymptomatic group compared with CEMRA (3 of 67) and DUS (1 of 66) (P=0.0186 versus MRA, P=0.0020 versus DUS). CONCLUSIONS: With the techniques as utilized in our study, the overall concordance rates of combined noninvasive methods are similar for measuring carotid stenosis in clinical routine practice, but in asymptomatic carotid stenosis, the decision making for surgery is significantly altered if DUS and CTA are considered in place of DUS and CEMRA. PMID- 14764933 TI - Anticardiolipin antibodies and risk of ischemic stroke and transient ischemic attack: the Framingham cohort and offspring study. AB - BACKGROUND AND PURPOSE: The role of anticardiolipin antibodies (aCLs) as novel risk factors for ischemic stroke and transient ischemic attacks (TIAs) has been a matter of debate. Prior cohort studies included only selected subjects, mostly men. We related serum concentrations of aCLs to incident first ischemic stroke/TIA among men and women in the Framingham Heart Study cohort and offspring. METHODS: There were a total of 2712 women (mean age, 59.3 years) and 2262 men (mean age, 58.3 years) free of stroke/TIA at the time of their baseline examinations. An enzyme immunoassay was used to measure aCLs. Optical density of the sample serum compared with the reference serum was defined as the aCL screening ratio (aCL SR). Analyses were based on sex-specific aCL SR quartiles and individual ratios. RESULTS: During the 11-year follow-up, 222 ischemic strokes/TIAs occurred. In multivariate analysis, after adjustment for age, prior cardiovascular disease, systolic blood pressure, diabetes, smoking, C-reactive protein, and total and high-density lipoprotein cholesterol levels, an aCL SR of >0.4 (78% of sample) was significantly associated with an increased risk of ischemic stroke/TIA for women (hazard ratio [HR], 2.6; 95% confidence interval [CI], 1.3 to 5.4; absolute risk, 3.2%, 95% CI, 2.2 to 4.3) but not in men (HR, 1.3; 95% CI, 0.7 to 2.4; absolute risk, 4.5%; 95% CI, 3.0 to 6.0). Similar results were obtained when the higher 3 aCL SR quartiles were compared with the lowest. CONCLUSIONS: Elevated serum concentrations of aCLs, independently of other cardiovascular risk factors, significantly predict the risk of future ischemic stroke and TIA in women but not in men. PMID- 14764934 TI - Isolation of functional RNA from small amounts of different grape and apple tissues. AB - An efficient, simple, and small-scale procedure for isolating functional ribonucleic acid (RNA) was successfully applied to many different tissues of grape and apple. These woody plants are rich in polyphenolic compounds and polysaccharides that could impair the RNA extraction. The method chosen is based on the use of hot borate buffer at alkaline pH supplemented with several adjuvants and followed by selective precipitations. Starting with only 0.4 g of fresh tissue and working with small tubes (2 mL), we were able to obtain good yields of high-quality RNA suitable for further applications. The procedure can be proposed for many applications, and it is particularly highly recommended when isolating RNA from a large number of samples. PMID- 14764935 TI - Cloning of the thermostable alpha-amylase gene from Pyrococcus woesei in Escherichia coli: isolation and some properties of the enzyme. AB - Pyrococcus woesei (DSM 3773) alpha-amylase gene was cloned into pET21d(+) and pYTB2 plasmids, and the pET21d(+)alpha-amyl and pYTB2alpha-amyl vectors obtained were used for expression of thermostable alpha-amylase or fusion of alpha-amylase and intein in Escherichia coli BL21(DE3) or BL21(DE3)pLysS cells, respectively. As compared with other expression systems, the synthesis of alpha-amylase in fusion with intein in E. coli BL21(DE3)pLysS strain led to a lower level of inclusion bodies formation-they exhibit only 35% of total cell activity-and high productivity of the soluble enzyme form (195,000 U/L of the growth medium). The thermostable alpha-amylase can be purified free of most of the bacterial protein and released from fusion with intein by heat treatment at about 75 degrees C in the presence of thiol compounds. The recombinant enzyme has maximal activity at pH 5.6 and 95 degrees C. The half-life of this preparation in 0.05 M acetate buffer (pH 5.6) at 90 degrees C and 110 degrees C was 11 h and 3.5 h, respectively, and retained 24% of residual activity following incubation for 2 h at 120 degrees C. Maltose was the main end product of starch hydrolysis catalyzed by this alpha-amylase. However, small amounts of glucose and some residual unconverted oligosaccharides were also detected. Furthermore, this enzyme shows remarkable activity toward glycogen (49.9% of the value determined for starch hydrolysis) but not toward pullulan. PMID- 14764936 TI - Evaluation of different conditions for ligating dumbbell-shaped oligonucleotides. AB - We tested three different standard ligation conditions (37 degrees C for 30 min, 16 degrees C for 24 h, and 4 degrees C for 48 h) to generate dumbbell-shaped oligonucleotides (ODNs) as transcription factor decoys for SOX9 and alphaA crystallin binding protein 1 (CRYBP1), which are positive and negative transcriptional regulators for type II collagen expression in chondrocytes. Decoy ODN for CRYBP1 was successfully produced as a "dumbbell" by all three conditions. A small amount of decoy ODN for SOX9, however, remained unligated under all three ligation conditions. Ligation at 4 degrees C for 48 h appeared to be the least desirable for SOX9 ODN. Transfection experiments with the SOX9 ODN ligated in different conditions and a luciferase-based reporter system also supports this conclusion. In general, shorter incubation time produced more acceptable results for this ODN than incubation for a longer time. These data suggest that different ligation conditions should be tested prior to creating dumbbell-shaped ODNs for transfection experiments. PMID- 14764937 TI - Infection by Shiga toxin-producing Escherichia coli: an overview. AB - Shiga toxin-producing Escherichia coli (STEC), especially of serotype O157:H7, cause a zoonotic food or waterborne enteric illness that is often associated with large epidemic outbreaks as well as the hemolytic uremic syndrome (HUS), the leading cause of acute renal failure in children. After ingestion, STEC colonize enterocytes of the large bowel with a characteristic attaching and effacing pathology, which is mediated by components of a type III secretion apparatus encoded by the LEE pathogenicity island. Shiga toxins are translocated from the bowel to the circularoty system and transported by leukocytes to capillary endothelial cells in renal glomeruli and other organs. After binding to the receptor globotriaosylceramide on target cells, the toxin is internalized by receptor-mediated endocytosis and interacts with the subcellular machinery to inhibit protein synthesis. This leads to pathophysiological changes that result in HUS. Specific therapeutic or preventive strategies are presently not available. The recent sequencing of genomes of two epidemic E. coli O157 strains has revealed novel pathogenicity islands which will likely provide new insights into the virulence of these bacteria. PMID- 14764938 TI - Designing safer (soft) drugs by avoiding the formation of toxic and oxidative metabolites. AB - Integration metabolic considerations into the drug-design process can allow safer pharmaceuticals to be designed. "Soft" drugs are designed to be deactivated in a predictable and controllable way after achieving their therapeutic role. They are designed to be metabolized rapidly and by avoiding oxidative pathways into inactive and nontoxic species. Successful application of such design principles has already resulted in a number of marketed drugs. The present article illustrates advantages inherent in avoiding the formation of oxidative metabolites, with examples that include soft bufuralol analogs and soft insecticides such as chlorobenzilate and malathion. Design principles for various soft drug classes are briefly summarized together with computerized tools intended to make the application of these principles more quantitative and more accessible. PMID- 14764939 TI - Pre-PCR processing: strategies to generate PCR-compatible samples. AB - Polymerase chain reaction (PCR) is recognized as a rapid, sensitive, and specific molecular diagnostic tool for the analysis of nucleic acids. However, the sensitivity and kinetics of diagnostic PCR may be dramatically reduced when applied directly to biological samples, such as blood and feces, owing to PCR inhibitory components. As a result, pre-PCR processing procedures have been developed to remove or reduce the effects of PCR inhibitors. Pre-PCR processing comprises all steps prior to the detection of PCR products, that is, sampling, sample preparation, and deoxyribonucleic acid (DNA) amplification. The aim of pre PCR processing is to convert a complex biological sample with its target nucleic acids/cells into PCR-amplifiable samples by combining sample preparation and amplification conditions. Several different pre-PCR processing strategies are used: (1) optimization of the DNA amplification conditions by the use of alternative DNA polymerases and/or amplification facilitators, (2) optimization of the sample preparation method, (3) optimization of the sampling method, and (4) combinations of the different strategies. This review describes different pre PCR processing strategies to circumvent PCR inhibition to allow accurate and precise DNA amplification. PMID- 14764941 TI - Rat liver-targeted naked plasmid DNA transfer by tail vein injection. AB - High levels of foreign gene expression in mouse hepatocytes can be achieved by "hydrodynamics-based transfection," the rapid injection of a large volume of a naked deoxyribonucleic acid (DNA) solution into the tail vein. Rats are more tolerant of the frequent phlebotomies required for monitoring blood parameters than mice and, thus, are more suitable for some biomedical research. Recently, we demonstrated that hydrodynamics-based transfection can also be used to deliver naked plasmid DNA into the normal rat, which is more than 10 times larger than the mouse. We performed the tail vein injection using a syringe with a winged needle equipped with an external tube. Injection of a lac Z expression plasmid, pCAGGS-lac Z by this technique resulted in the exclusive detection of beta galactosidase in the liver. We also injected a rat erythropoietin (Epo) expression plasmid, pCAGGS-Epo (800 microg). Maximal Epo gene expression was achieved when a 25-mL injection volume (approx 100 mL/kg body wt) was transferred within 15 s. PMID- 14764940 TI - MALDI-TOF mass spectrometry: a versatile tool for high-performance DNA analysis. AB - Matrix-assisted laser desorption/ionization (MALDI) time-of-flight (TOF) mass spectrometry (MS) has developed during the past decade into a versatile tool for biopolymer analysis. The aim of this review is to summarize this development and outline the applications, which have been enabled for routine use in the field of nucleic acid analysis. These include the analysis of mutations, the resequencing of amplicons with a known reference sequence, and the quantitative analysis of gene expression and allelic frequencies in complex DNA mixtures. PMID- 14764942 TI - Effects of probucol on renal function and urinary protein excretion in spontaneously hypercholesterolemic rats fed a normal or high cholesterol diet. AB - BACKGROUND/AIM: Spontaneously hypercholesterolemic (SHC) rats develop hypercholesterolemia and focal glomerular sclerosis, and have been thought to be a model of lipid-induced glomerular injury. However, recent studies suggest that the hypercholesterolemia might be due to secondary mechanisms by massive proteinuria. The purpose of the present study was to determine in SHC rats the effects of a high cholesterol diet on serum lipid profiles and renal function/histology, and to examine whether or not the model of lipid-induced renal injury could be developed in a short period of the time. The effects of probucol were also studied. METHODS: SHC rats were fed a high cholesterol diet for 6 weeks (H) or with probucol (HP), while control SHC rats were fed normal rat chow (N) or with probucol (P). Lipid profile and renal function/histology were examined. RESULTS: H and HP showed increased levels of urinary protein excretion and serum creatinine, as well as extremely high serum cholesterol levels, compared with N and P. HP tended to show reduced urinary protein excretion compared with H, but the difference was not statistically significant. H and HP presented histologically characteristic changes with numerous foam cells accumulated in the glomerular mesangial area, and showed glomerular sclerosis. CONCLUSION: The data demonstrate that SHC rats have an intrinsically abnormal lipid metabolism, and that probucol does not exert obviously beneficial effects on renal function or lipid-lowering action. A lipid-induced renal injury model of rats was produced in 6 weeks. PMID- 14764943 TI - The seasonal variation in allergic rhinitis and its correlation with outdoor allergens in Kuwait. AB - BACKGROUND: Allergic rhinitis (AR) is the most common allergic problem in Kuwait. Most of the patients who have either AR or asthma are referred to the Al-Rashed Allergy Center. OBJECTIVE: To determine if there is a seasonal variation in AR in Kuwait and to correlate it with the daily pollen count. METHODS: Information about the new patients referred to the center over a 5-year study period (1996 2000) was extracted from the center's records. The daily pollen count in Kuwait city was obtained from the Air Biology Laboratory. RESULTS: There was a significant seasonal variation with a bimodal increase in the number of patients with AR referred to the center. The main peak in the number of patients occurred in September-October, and there was a smaller peak in April-May. The mean number +/- SD of new patients per month over the 5-year period varied from 87 +/- 32 for December to 367 +/- 104 for September. Similarly, the average daily pollen count varied from 3.7 +/- 1.0 pollens per mm3 in January to 124 +/- 92 in October. There was high correlation between the number of new AR patients and the average total pollen count (Pearson correlation, r=0.77, p<0.001), as well as with Chenopodiaceae and Amaranthaceae (weed) pollens (r=0.75, p<0.001), while there was no correlation between the number of new patients and either tree or grass pollens. CONCLUSION: Seasonal AR occurs during two periods in Kuwait, i.e. September-October and April-May, with September-October being the main season. The rise in AR during late summer in Kuwait is mainly associated with the pollination of Chenopodiaceae species. PMID- 14764944 TI - Wheat allergy: clinical and laboratory findings. AB - BACKGROUND: Food allergy affects 6-8% of infants and wheat allergy is one of the common food allergies among children. The clinical and laboratory manifestations of wheat allergy were evaluated in this study. METHODS: Thirty-two children (< or =12 years old) with suspected wheat allergy were evaluated for wheat allergy. The patients underwent wheat skin prick test (SPT), measurement of wheat-specific IgE and wheat challenge test. The patients with a convincing history of anaphylaxis following ingestion of wheat or with a positive challenge test, and those with a history of immediate hypersensitivity reaction following ingestion of wheat in addition to a positive wheat SPT and/or positive wheat-specific IgE were considered wheat allergic. Then, the laboratory and clinical manifestations of their disease were studied. RESULTS: Among patients with suspected wheat allergy, 24 patients with definite wheat allergy were identified. Anaphylaxis was a dominant clinical feature, accounting for 54.1% of acute symptoms. Chronic allergy symptoms like asthma and eczema were noted in 50% of the patients. Wheat specific IgE was higher in patients with anaphylaxis (p<0.02) and the risk of anaphylaxis was 14.4 times more in patients with wheat-specific IgE equal to or more than 3+. CONCLUSIONS: Anaphylaxis had occurred in a remarkable number of patients repeatedly, which demonstrates the severity of the reactions, poor knowledge of the disease and probable existence of more patients with mild reactions. Regarding the higher level of wheat-specific IgE in patients with anaphylaxis, wheat-specific IgE could be used to predict the severity of symptoms. PMID- 14764945 TI - Allergy and tumour outcome after primary cancer therapy. AB - BACKGROUND: Over the last decade several papers have dealt with the possible interference of allergies in both the infectious disease incidence and tumour development. In the light of all these observations we analysed several tumour patients for a possible interaction between a state of allergy and tumour development and progression after primary cancer therapy. METHODS: This study included 1,055 patients with different types of solid tumours admitted consecutively between 1994 and 2002 to the Cagliari University Polyclinic. After primary surgery or medical therapy (or both), 92 allergic subjects and 182 non allergic patients were studied over a follow-up period of 6-96 months (median 23). RESULTS: Among 1,055 tumour-bearing patients, the prevalence of allergy was found to be about 8% versus 16-37% in a population of non-tumour-bearing subjects. After primary cancer therapy, allergic patients turned out to have a 20% higher probability of being cured and about a 50% lower risk of tumour progression as compared to non-allergic ones. The observed differences were statistically significant (p=0.013). CONCLUSIONS: On the basis of our findings, we suggest that allergic subjects seem to have a better prognosis than non allergic ones for disease outcome after cancer therapy. PMID- 14764946 TI - Discovery and immunologic validation of new antigens for therapeutic cancer vaccines. AB - Immunotherapy using both active and passive approaches is increasingly being used as a modality to treat human cancer. The last decade has seen a tremendous burst of activity in antigen discovery in cancer, and many new targets have now been identified for both monoclonal antibody therapy and active immunization. In addition, advances have been made in our understanding of the immune response against cancer and how new vaccine vectors, such as poxviruses, alphaviruses and bacterial vectors, can be used to overcome some of the traditional hurdles (e.g. self-tolerance and immune suppression in cancer patients) that have hindered the generation of effective antitumor immune responses. Improvements in genomics technology in the area of DNA microarrays and differential display and subtractive hybridization together with a new wave of mass spectrometry-based proteomics tools, as well as more sensitive assays to validate the immunoreactivity of new antigens, have all accelerated the rate of new antigen discovery in cancer. This rapid progress should initiate a major paradigm shift in how we treat cancer within the next 10 years, where, instead of being a novelty, the combination of targeted T cell-based vaccines and antiangiogenesis therapies will be routinely combined with traditional chemotherapy. The successful combination of these approaches will change the face of cancer from a relatively acute, life-threatening disease to a manageable chronic disorder with long survival times. PMID- 14764947 TI - Glucoreceptors located inside the blood-brain barrier mediate hypoglycemia induced LH inhibition in man. AB - OBJECTIVE: To establish the role of hyperinsulinemia and hypoglycemia during the insulin tolerance test (ITT) in the regulation of luteinizing hormone (LH) secretion and the location with respect to the blood-brain barrier (BBB) of the glucosensitive areas controlling LH release. METHODS: The LH-secretory pattern during an ITT (0.15 IU/kg body weight) was evaluated in 8 normal men during infusion with normal saline (control test), glucose or fructose. RESULTS: lnsulin induced hypoglycemia produced a significant decrement in serum LH levels in the control test, but not when the concomitant infusion of glucose prevented hypoglycemia. Fructose infusion did not change LH decrease during ITT. CONCLUSIONS: These data exclude a direct role of hyperinsulinemia in the mechanism underlying the inhibition of LH secretion during ITT. Furthermore, since glucose but not fructose crosses the BBB, the LH decrease during ITT appears to be generated by hypoglycemia at the level of glucosensitive areas located inside the BBB. PMID- 14764948 TI - Hypoketotic hypofattyacidaemic hypoinsulinaemic hypoglycaemia in a child with hemihypertrophy? A new syndrome. AB - BACKGROUND: Recurrent and persistent hypoketotic, hypofattyacidaemic hypoglycaemia in infancy and childhood is most frequently due to hyperinsulinism of infancy. This biochemical profile can also be due to non-islet cell tumour hypoglycaemia or circulating insulin-receptor autoantibodies. Hyperinsulinaemic hypoglycaemia is also seen in children with the Beckwith-Wiedemann syndrome, where it is usually transient. METHODS/RESULTS: We report a novel case of child with hemihypertrophy and severe persistent hypoketotic, hypofattyacidaemic hypoinsulinaemic hypoglycaemia. No 'big' pro-IGF2 forms or circulating insulin receptor antibodies were found. Glucose and protein isotope turnover studies showed marked suppression of hepatic glucose production during fasting. There was no evidence for constitutive autophosphorylation of the insulin or IGF-1 receptor, and no evidence for up-regulation of IGF-1 receptor. CONCLUSION: The precise pathophysiology of this novel case is still unclear. PMID- 14764949 TI - The coronary effects of parathyroid hormone. AB - OBJECTIVE: The aim of the present study was to characterize the role of the ATP sensitive potassium channels (K(+)(ATP)) in the coronary dilator action of parathyroid hormone (PTH). METHODS: Dose-response curves of intracoronary administrated PTH (0.15-1.33 nmol) were obtained in control phases and during continuous intracoronary administration of the K(+)(ATP) channel-selective antagonist glibenclamide (0.1-1.0 micromol/min) in dogs (n = 13). RESULTS: Increments of integrated coronary conductance (excess coronary conductance) at PTH doses of 0.15 and 1.33 nmol were 1.17 versus 0.03 ml/mm Hg (p < 0.05) and 4.03 versus 0.94 ml/mm Hg (p < 0.05) in the control versus during maximal blockade, respectively. CONCLUSION: The results indicate that the activation of K(+)(ATP) channels significantly contributes to the PTH-induced coronary vasodilation. PMID- 14764950 TI - Longitudinal data for intrauterine levels of fetal IGF-I and IGF-II. AB - OBJECTIVE: An increasing body of evidence supports a major role for the insulin like growth factors (IGFs) in the control of human fetal growth. Individual data at various times of pregnancy suggest that IGF-I and IGF-II levels remain stable up to the 33rd week of pregnancy. Thereafter, both increase to reach values 2-3 times higher at term. In order to provide an accurate reflection of fetal IGFs in utero, we sampled fetal blood from the umbilical cord by cordocentesis. METHODS: We measured IGF-I and IGF-II in 12 fetuses longitudinally for up to 5 times between the 21st week of gestation and delivery. RESULTS: All patients showed a progressive increase in IGF-I and IGF-II levels. Data determined during different time intervals (before 29th, 29th to 32nd, after 32nd week) were compared and the main increase was found after the 32nd week. The median for IGF-I before the 29th week was 33.5 ng/ml (range 19-40.5) and increased to 41 ng/ml (32-59) between the 29th to 32nd and further to 54.1 ng/ml (range 17-70) thereafter. During the same time interval, the median for IGF-II increased from 217 ng/ml (86-326) to 349 ng/ml (227-467). In 7 patients, cord blood after delivery was available. For IGF II a further increase was consistently found after birth (from 282 ng/ml (175 511) to 393 ng/ml (297-513)), whereas only 2 fetuses showed an increase in IGF-I. CONCLUSION: We conclude that in human fetuses, IGF-I and IGF-II levels increase longitudinally throughout pregnancy. Therefore, they may become important markers of healthy fetal development. PMID- 14764951 TI - Comments on ethics of experimental therapies. PMID- 14764952 TI - Stem cell transplantation and ethics: a European overview. AB - Stem cell transplantation requires a source of stem cells. The potential use of human embryonic stem cells for research and therapy has created major ethical and political debates in Europe. The first section of this paper gives an overview of the official regulatory responses to the possibility of stem cell transplantation in a range of European jurisdictions. It focuses on the regulation of: (1) the use and production of embryonic stem cells, (2) the use of adult stem cells, and (3) the patenting of stem cell lines and products. The second section tries to unravel why regulatory responses have differed between different European countries, especially with regard to embryonic stem cells and focuses on two possible explanations for the differences: (1) ethical disagreement and (2) maintenance of regulatory consistency. It is argued that an exclusive focus on ethical disagreement cannot fully explain the differences in regulatory response, and that an understanding of the specific way in which regulations develop over time in each jurisdiction is crucial. Certain types of ethical consistency arguments can, for instance, only be mounted successfully if the jurisdiction in question already allows other forms of experiments involving human embryos. In the final section it is argued that the European regulatory developments in the future may be crucially dependent on whether actual stem cell products are developed that can potentially be traded across borders. PMID- 14764953 TI - Sources of stem cells: ethical controversies and policy developments in the United States. AB - The bioethical controversy about public policy in the United States regarding human stem cell research, particularly with regard to several potential sources of stem cells, including aborted fetuses, unimplanted embryos following in vitro fertilization (IVF), and embryos created through IVF or cloning for research purposes is examined. On the one hand, the debate about the use of federal funds for this research was not resolved by President Bush's policy, announced in August 2001, because only a few cell lines are available under this restrictive policy. On the other hand, debate persists about whether the US should adopt a prohibitive, regulatory, or permissive policy toward the deliberate creation of embryos for biomedical research through somatic cell nuclear transfer. Although there is a wide consensus about the need to ban cloning-for-reproduction, there is deep division, reflected in the work of the President's Council on Bioethics, about a possible ban on cloning-for-biomedical research. The author concludes that ethical controversy will probably persist about both types of policy, and that the policy stalemate will probably also continue, because of fundamental disagreements about the moral status of the early embryo. PMID- 14764954 TI - Ethical questions raised by in utero therapeutics with stem cells and gene therapy. AB - The plasticity of living beings is limitless and our guidelines are constantly upset. Several ethical questions arise with this new in utero therapeutic approach: (1) The possibility of a mother giving truly informed consent when she is divided between a proposal to terminate her pregnancy and resignation to the birth of an abnormal child. How can she choose between three therapeutic proposals, when two of them are unbearable and only one carries some hope? (2) If a dead fetus is used as a donor of hepatic or hematologic stem cells, should this be based on the mother's consent? If so, what status is given to this fetus? Is it possible to take into consideration at the same time the growing demand to recognize the fetus as a person and his gift? (3) It is important to separate the services that receive dead fetuses from those that perform the injection of fetal stem cells so that the fetus does not become a mere therapeutic tool. (4) Even if the technique is not very difficult, can ex vivo transduction of mesenchymatous cells be performed as a kind of gene therapy without causing great anxiety over the embryo's germinal future? The main question is that even though surgery and medical therapeutics for the fetus have made great progress and become commonplace, the use of stem cell transplantations and eventually gene therapy privileges experimental medicine and research that are more and more difficult to conceive on an ethical level. If indeed fetal therapeutics seems to hold out promise for the future, the very speed with which we pass from conception to its experimental realization with a child must necessarily challenge our thinking. PMID- 14764955 TI - Prenatal diagnosis of Apert syndrome. AB - OBJECTIVE: The role of the human fibroblast growth factor receptor (FGFR) gene family in current prenatal diagnosis and management of craniosynostosis syndromes and skeletal dysplasias is discussed. METHOD: We present the antenatal ultrasound findings, diagnosis, and management of 2 cases of Apert syndrome before and after molecular prenatal diagnosis was available. RESULTS AND CONCLUSION: Discovery of mutations in FGFR genes now allows the definitive antenatal diagnosis of Apert syndrome, other craniosynostosis syndromes, and skeletal dysplasias. PMID- 14764956 TI - Congenital visceral malformations--role of perinatal autopsy in diagnosis. AB - OBJECTIVES: Congenital malformations are one of the leading causes of perinatal deaths and infant mortality. The objective of the present study is to detect visceral malformations in perinatal autopsies. DESIGN: A retrospective analysis of perinatal autopsies performed between 1998 and 2001 was done. Various visceral malformations were noted and categorized as urologic, cardiac, respiratory, gastrointestinal and miscellaneous. RESULTS: Out of a total of 62 perinatal autopsies performed, congenital malformations were present in 38.7% of cases. Visceral malformations were observed in 24.1% of cases. Urologic malformations were the commonest (14.1%), followed by cardiac (8%) malformations. Associated external malformations were present in 6/15 cases, cardiac malformations being commonly associated with skeletal malformations. CONCLUSIONS: In all the cases, internal malformations were not suspected clinically. Thus, autopsy is an invaluable tool for detecting visceral malformations, adding to the clinical diagnosis and providing a feedback to the parents. PMID- 14764957 TI - Characteristic findings for diagnosis of baby complicated with both the VACTERL association and duodenal atresia. AB - The VACTERL [vertebral defects (V), anal atresia (A), cardiac anomaly (C), tracheal-esophageal fistula with esophageal atresia (TE), renal defects (R), and radial limb dysplasia (L)] association can sometimes be diagnosed by ultrasonography and magnetic resonance imaging (MRI). Although the preaxial limb anomalies on ultrasonography were strongly associated with VACTERL association, the rate of limb anomalies is low. On ultrasonography, useful findings for prenatal diagnosis are a combination of esophageal atresia with hydramnion and renal anomalies. If esophageal atresia cannot be detected due to masking, diagnosis may be very difficult. In this case report, we reported the VACTERL association along with duodenal atresia. The detection of characteristic findings (enlarged stomach and duodenum, possibly change in gallbladder) by use of ultrasonography and MRI might be useful for the prenatal diagnosis of such cases. PMID- 14764958 TI - Genetic amniocentesis in multiple pregnancy. AB - OBJECTIVES: Second-trimester genetic amniocentesis is the most frequently used invasive prenatal diagnostic technique. Several reports have been published about the effect of genetic amniocentesis on fetal loss in multiple pregnancies over the past two decades. Here we analyze our experience with genetic amniocentesis in multiple pregnancies over the past 10 years. METHODS: Details of 184 multiple pregnancies were processed in all cases in whom genetic amniocentesis was performed in women who presented at our department since 1990. The outcomes of 175 cases (95.1%) out of 184 genetic amniocenteses were available to us. As a control group, we followed up the outcome of 300 twin pregnancies in which no genetic amniocenteses were performed. RESULTS: We found that the proportion of spontaneous losses in multiple pregnancies between the 18th and the 24th gestational weeks was 2.39%, whereas if genetic amniocentesis was performed the loss rate before the 24th week was 3.87%. The perinatal mortality rate was 10.03/1,000 in the group who underwent amniocentesis, while it was 10.52/1,000 in the group without amniocentesis. CONCLUSIONS: Our results suggest that the genetic amniocentesis performed in multiple pregnancies slightly increased (1.48%) the fetal loss rate until the 24th week. Beyond 5 weeks after the procedure, no consequent fetal loss should be expected. In our study the intervention did not have any undesired effect on perinatal mortality rates. PMID- 14764959 TI - Effect of smoking on serum concentrations of total homocysteine, folate, vitamin B12, and nitric oxide in pregnancy: a preliminary study. AB - OBJECTIVE: Nitric oxide (NO) is a potent vasodilator released by endothelial cells that plays an important role in modulating maternal and fetal vascular tone in normal pregnancy. Lower plasma levels of vitamins may result in hyperhomocysteinemia, a known risk factor in pregnancy. The aim of this study was to investigate whether there are alterations in the serum levels of total homocysteine (tHcy), folate, vitamin B12, and total nitrite, as an index of NO, in smoking as compared with age-matched nonsmoking pregnant women. METHODS: Thirty-three women (19 smoking and 14 nonsmoking) between 16 and 22 weeks of their gestation were included in this study. The serum tHcy levels were analyzed by using an enzyme-linked immunosorbent assay kit. Vitamin B12 and folate values were measured by means of DPC kits. Total nitrite was measured by Griess reaction as an index of endogenous NO production. RESULTS: The serum tHcy concentrations were significantly increased in smoking as compared with nonsmoking pregnant women (p<0.001). The folate and vitamin B12 concentrations were lower in smoking than in nonsmoking pregnant women, but only the differences in folate concentrations were statistically significant (p<0.001). The tHcy concentrations showed a significant negative correlation with folate in the smoking pregnant women. The serum total nitrite concentrations were lower in smoking than in nonsmoking pregnant women (p<0.05). In addition, the serum nitrite levels in smoking pregnant women had significant negative correlations with tHcy and positive correlations with folate and vitamin B12 levels. CONCLUSIONS: In the light of our findings, we propose that smoking might enhance the vasoconstrictor capacity in pregnant women by increased tHcy concentrations and by a simultaneous decrease in the production of NO which is a vasodilator compound. PMID- 14764960 TI - Dysthymic reactions of women undergoing chorionic villus sampling for prenatal diagnosis of hemoglobinopathies or karyotyping. AB - OBJECTIVES: The aim of the study was to investigate the levels of anxiety and depression, as well as liability to depression, of women undergoing chorionic villus sampling (CVS) due to the risk of giving birth to a child suffering from hemoglobinopathy, or trisomy. METHODS: The study population consisted of 309 women who attended the Fetal Medicine Unit of Alexandra Hospital, University of Athens, Athens, Greece for a first trimester chorionic villus sampling (CVS). One hundred and fifty-nine women (group A) underwent CVS due to increased possibility of carrying an embryo with beta-thalassemia, while 150 women had the procedure because of an increased nuchal translucency measurement, or a positive first trimester biochemical screening (group B). Three hundred and nine women, matched by age and gestational age with those of the study group, who were not subjected to any diagnostic intervention formed the control group. We further investigated differences of dysthymic reactions among three subgroups of women at risk for hemoglobinopathy: (a) women undergoing their first pregnancy; (b) women who had been pregnant more than once but had not any children, and (c) women who had given birth to at least one healthy child, irrespective of previous terminations. RESULTS: The incidence of clinically elevated levels of anxiety and depression was significantly higher in groups A and B compared to controls (p<0.001), while no significant difference was found in mean anxiety and depression, as well as the liability to depression among the three groups. Clinically elevated levels of depression were found in 10.1 and 4.7% of the women of the hemoglobinopathy and karyotyping group, respectively. Especially women who had no children, due to previous pregnancy terminations presented significantly higher incidence of elevated depression compared to women who were pregnant for the first time, or women who had given birth to at least one healthy child in their life. CONCLUSIONS: The findings of this study highlight the necessity of training medical and nursing personnel to understand and respond to the psychological and social needs of women undergoing CVS due to increased risk for hemoglobinopathy, and especially to those who have no children and who have a history of pregnancy termination due to an affected fetus. PMID- 14764961 TI - Prenatal diagnosis of a 'minor' form of Brachmann-de Lange syndrome by three dimensional sonography and three-dimensional computed tomography. AB - Brachmann-de Lange syndrome is a congenital disease characterized by severe mental retardation, pre- and postnatal symmetric growth delay, limb defects, visceral anomalies, hirsutism, and a typical face. The authors describe the prenatal sonographic pattern of Brachmann-de Lange syndrome suspected at 20 weeks of gestation, with severe intrauterine growth retardation, facial dysmorphism, cardiac abnormality, and micromelia without the typical defects of the upper limbs. Fetal karyotyping was normal. The diagnosis of a 'minor' form of Brachmann de Lange syndrome was confirmed at 28 weeks of gestation by using three dimensional sonography in order to assess precisely the facial dysmorphism and by performing a three-dimensional computed tomography of the upper limbs in order to identify the subtle abnormalities of the radial head and of the first metacarpal bone. PMID- 14764962 TI - Prenatal maternal stress, prenatal fetal movements and perinatal temperament factors influence behavior and school marks at the age of 6 years. AB - Niederhofer showed in 1994 that maternal stress during pregnancy seemed to be associated with temperament in early childhood. This study examined the correlation between maternal stress during pregnancy in 227 women, ultrasound observation of intrauterine fetal movements, the child's temperament in early childhood, and the child's school marks at the age of 6 years. Our results show a significant correlation between prenatal maternal stress, perinatal temperament of the child and his/her school marks at the age of 6 years. Only intrauterine fetal movements were associated neither with stress during pregnancy nor with early personality. PMID- 14764963 TI - Antenatal diagnostic aspects of unilateral multicystic kidney dysplasia- sensitivity, specificity, predictive values, differential diagnoses, associated malformations and consequences. AB - OBJECTIVES: Unilateral multicystic kidney dysplasia (MCKD) is the second most common urinary tract abnormality diagnosed antenatally. Whilst an isolated unilateral MCKD has a good prognosis, a poor outcome must be expected when MCKD is associated with other complex abnormalities. MATERIAL: Out of 11,176 cases, 693 fetuses were suspected of having urinary tract abnormalities. Urological findings were confirmed in 548 of them. Unilateral cystic kidney was diagnosed prenatally in 85 cases. RESULTS: The study results in a total of 107 cases with proven MCKD. Eighty-five pregnancies with a prenatal diagnosis of MCKD were analysed. The antenatal diagnosis of MCKD was confirmed in 56 cases. Fifty-one children were found to have unilateral MCKD where this had not been explicitly suspected from antenatal scanning. CONCLUSION: Unilateral MCKD is a malformation with an excellent prognosis for child survival and global renal function if encountered in isolation. However, our analysis of live infants and autopsy cases demonstrates a high proportion of severe associated malformations of the urinary tract and other organ systems. PMID- 14764964 TI - The peritoneal route as a safe pathway for early in utero therapies: illustration by a 12-year follow-up after conservative management of severe Rhesus allo immunization. AB - OBJECTIVE: We report a case of an extremely severe Rhesus allo-immunization treated very early in pregnancy 12 years ago. METHODS AND RESULTS: After chorionic villus sampling at 12 weeks for fetal blood phenotyping, two intraperitoneal transfusions at 14 and 15 weeks were given followed by two intravascular and seven exchange transfusions. A girl weighing 2,940 g was delivered vaginally at term after external cephalic version for breech presentation. To date her neurological and social development is normal. CONCLUSIONS: Since the success of haematopoietic stem cell transplantations for the treatment of congenital haematologic diseases could imply early and repetitive procedures, this observation enlightens the technical feasibility of such an invasive approach and its relative safety for subsequent development. PMID- 14764965 TI - A comparison between maternal serum free beta-human chorionic gonadotrophin and pregnancy-associated plasma protein A levels in first-trimester twin and singleton pregnancies. AB - OBJECTIVES: To determine the levels of first-trimester free beta-human chorionic gonadotrophin (free betahCG), pregnancy-associated plasma protein A (PAPP-A) and nuchal translucency (NT) in twin pregnancies. METHODS: The study included 93 patients with twin pregnancy and 4,977 with singletons who underwent first trimester testing using free betahCG, PAPP-A and NT at 10-13 weeks of gestational age. RESULTS: In twin pregnancies, the maternal serum free betahCG level was 2.18 higher and the PAPP-A level was 2.38 higher than in singleton pregnancies. These marker levels were significantly higher than the expected 2.0 multiples of the median (MoM). In contrast, NT values did not differ between twins and singletons. CONCLUSION: These data may be used to establish first-trimester combined screening for twin pregnancies. PMID- 14764966 TI - Pericardial hemangioma presenting as thoracic mass in utero. AB - Pericardial hemangiomas are rare lesions. We present the case of an infant who was referred to our fetal diagnosis and treatment group for the presence of a left thoracic mass, pleural effusion, and mediastinal shift on fetal ultrasound. The characteristics of the lesion suggested the presence of a pulmonary sequestration. A chest radiograph done at birth was normal. At 2 weeks of age, an enhancing lesion of the left pericardium was identified on chest CT. A cardiac MRI demonstrated enhancement of the mass on T2-weighted images. The patient underwent thoracoscopic assessment of the mass for diagnostic purposes. Multiple lesions were identified along the left pericardium and diaphragm. A frozen section biopsy revealed a hemangioma. The natural history for hemangiomas is gradual regression; however, they may increase acutely in size and cause symptoms prior to involution. Investigations should be performed to identify the involvement of other organs. This case illustrates the need to closely follow all patients with prenatally diagnosed thoracic masses with CT imaging, even when they are asymptomatic and have a normal chest radiograph at birth. PMID- 14764967 TI - Fetal hemolytic disease due to anti-Rh17 alloimmunization. AB - OBJECTIVE: To delineate clinical features of a case of fetal hemolytic disease due to anti-Rh17, along with a review of relevant studies published in English and Japanese. METHODS: We present clinical features of a -D-/-D- phenotype woman with anti-Rh17 alloimmunization during pregnancy. Relevant English literature in the MEDLINE database was reviewed, while Japanese studies were searched in the Japana Centra Revuo Medicina database. RESULTS: A Japanese -D-/-D- woman with anti-Rh17 (Hro) was treated during pregnancy. Serial ultrasonography, antibody titers, amniocenteses, and cordocenteses were conducted for perinatal management. Amniocentesis results demonstrated a high delta optical density level of 450 in the amniotic fluid, while cordocentesis revealed alloimmunization between the mother and the fetus as well as fetal hemolytic anemia. Blood flow velocity in the middle cerebral artery indicated a rapid development of fetal anemia. The newborn demonstrated severe anemia and hyperbilirubinemia, which were successfully treated with exchange transfusions. Two cases of prenatally diagnosed fetal hemolytic disease due to anti-Rh17 were found published in English and 5 in Japanese. CONCLUSION: A -D-/-D- phenotype patient with anti-Rh17 was successfully managed during pregnancy and a good outcome for the neonate was achieved. Our results and a review of related literature led to the following suggestions. The first pregnancy in a -D-/-D- woman may be affected, an anamnestic immune response can easily occur during pregnancy, the level of anti Rh17 titer is indicative of the degree of fetal hemolysis, and appropriate intrauterine intervention is warranted for achievement of a good outcome. PMID- 14764968 TI - First-trimester findings associated with twin reversed arterial perfusion sequence. AB - INTRODUCTION: Regarding its pathogenesis, discordant development in early gestation, as well as vascular anastomoses between twins are postulated to be required for the establishment of the twin-reversed arterial perfusion (TRAP) sequence. However, first trimester findings associated with this complication have not yet been reported. CASE: A discordant monochorionic twin was revealed upon examination of a 24-year-old primigravida at 11 weeks' gestation. Cystic masses were identified on the back of the smaller twin, later followed by the appearance of skin edema and pericardial effusion, indicating cardiac failure. Subsequently, despite diagnosis of fetal demise at 15 weeks the lower body was shown to have further developed and the heartbeats appeared again, resulting in an acardia anceps or hemicardia. No remarkable change was observed in the larger normal twin. CONCLUSION: This occurrence was considered consistent with the current hypothesis regarding the pathogenesis of the acardiac anomaly. First trimester discordancy in a monochorionic twin gestation is considered to represent an early manifestation of TRAP sequence. PMID- 14764969 TI - Intrauterine therapy for the acutely enlarging fetal cystic hygroma. AB - Enlarged fetal cystic hygroma is known to cause life-threatening complications such as fetal hydrops and neonatal respiratory difficulty. A 28-year-old Japanese woman, gravida 0, presented with fetal cystic hygroma at 23 weeks of gestation. There were no other structural malformations or hydrops detected by ultrasonographic examination. In addition, the karyotype was diagnosed as normal through amniotic fluid analysis. The cystic lesion showed acute enlargement and intrauterine sclerotherapy using OK-432 was performed at 26 weeks. The size of the cyst initially decreased, which was followed by a gradual increase. A viable 3,098 g male infant was delivered by cesarean section at 37 weeks without any other complications. The infant had no clinical difficulty during the neonatal period and later underwent a surgical removal of the remaining cystic lesion. Cases of fetal cystic hygroma showing acute enlargement without other complications are considered good candidates for intrauterine therapy to prevent subsequent complications. PMID- 14764970 TI - Maternal mortality following diagnostic 2nd-trimester amniocentesis. AB - We present 2 cases of maternal mortality after transabdominal amniocentesis performed during the 2nd trimester of pregnancy. In both these cases, blood cultures revealed Escherichia coli. Broad-spectrum intravenous antibiotic treatment started immediately after admission to the hospital did not change the rapid progression of the disease. Despite evacuation of the uterus within <10 h from the diagnosis of septic abortion and transfer to the intensive care units to treat multiorgan failure, these patients died. Septic abortion and septic shock following transabdominal amniocentesis are very rare; however, they carry a serious risk to the patients' life. The combination of fever and leukopenia several days after amniocentesis should alert the physician to the evolution of sepsis. Because of the risk involved, information given to the patient prior to amniocentesis should refer to possible fetal complications and to the remote possibility of maternal risks as well. PMID- 14764971 TI - Multiplex nested PCR for preimplantation genetic diagnosis of spinal muscular atrophy. AB - OBJECTIVE: Spinal muscular atrophy (SMA) is a common autosomal recessive neuromuscular disorder caused in most patients by homozygous deletion of the SMN1 gene. For a carrier couple at a 25% risk of affected offspring, preimplantation genetic diagnosis (PGD) offers an alternative to prenatal diagnosis and termination of affected pregnancies. Our objective was to develop an accurate and reliable single-cell multiplex nested PCR analysis for PGD of SMA. METHODS: The method was developed on single blood leukocytes, obtained from healthy controls and an adult SMA type III patient with a known homozygous deletion of SMN1 exon 7 and 8. Multiplex nested PCR on single cells was used to co-amplify exons 7 and 8 of SMN. Additional multiplexing was performed with the ZFX/ZFY gene for sexing. Following successful establishment of the multiplex nested PCR protocol in single leukocytes, the technique was employed for PGD in 4 patients for a total of 7 cycles. In 2 patients, sexing was simultaneously performed using ZFX/ZFY. RESULTS: 220 single leukocytes from a normal individual and 220 from an SMA patient were analyzed. Exon 7 of SMN1 was amplified in 99% of normal single leukocytes and in none of the SMA-affected leukocytes. Exon 7 of SMN2 was amplified in 100% of both normal and SMA-affected leukocytes. Exon 8 of SMN1 was amplified in 98% of normal cells and in none of the SMA-affected leukocytes. Exon 8 of SMN2 was amplified in 96% of both normal and SMA-affected leukocytes. Amplification efficiency was 99% for ZFX/ZFY. There were no false-negative results and no contamination was detected in all wash-drop blanks tested. Seven PGD cycles were performed in 4 SMA-carrier couples with successful molecular analysis of 34 embryos and a total of 15 normal embryos transferred in 7 cycles. One clinical pregnancy has resulted in the delivery of a healthy male. Amniocentesis performed at 17 weeks confirmed the correct diagnosis for both SMA and sexing. CONCLUSIONS: These results suggest that our multiplex nested PCR protocol offers an efficient and accurate method for PGD of SMA while enabling the simultaneous analysis of an additional loci. PMID- 14764972 TI - Anal fistulotomy with radiofrequency. PMID- 14764974 TI - Genetic polymorphism of the renin-angiotensin system on the development of primary vesicoureteral reflux. AB - BACKGROUND: The familial clustering of vesicoureteral reflux (VUR) has suggested a genetic basis. This study was designed to investigate the genetic polymorphism of the renin-angiotensin system (RAS) in Korean children. METHODS: Genetic polymorphism of angiotensin-converting enzyme (ACE) and angiotensin II receptor genes was evaluated in 67 primary VUR patients and compared to 58 controls with no urological abnormalities. To detect the relation of the risk factors of primary VUR with the genetic polymorphism, the distribution of ACE, AT1 and AT2 genotypes after stratification by risk factors was also studied in the primary VUR patients. RESULTS: The incidence of AT2 A-1332G transition was significantly lower in primary VUR patients (p = 0.047). Furthermore, in the case of combination of ACE and AT2 gene, a significantly lower incidence of primary VUR was seen with II genotype of ACE and A-1332G transition in the AT2 receptor gene (p = 0.003). Concerning the risk factors of primary VUR, there were no biologically significant results. CONCLUSIONS: These findings indicate that a lower incidence of AT2 A-1332G transition is seen in primary VUR patients, at least in the Korean population. Also, in the case of combination of ACE and AT2 gene, the combination of ACE II genotype and AT2 A-1332G transition occurs infrequently in primary VUR. PMID- 14764975 TI - Nutrition of burned patients. AB - Burns form 5-12% of all traumas. About 2,200 of patients are annually hospitalized in Lithuania. In most cases people of the employable age get burned. The treatment is often long-lasting, and afterwards recovered patients often have invalidity from burn sequels. The mortality of hospitalized burned patients is about 10%. The most common causes of death are pulmonary edema, pneumonia, sepsis and multiorgan failure. All these complications are related with insufficient nutrition. These complications are extremely frequent and dangerous for patients with more than 20% of body burned. The nutritional support of burned patient gives a possibility to increase the survival probability, to decrease complication rate and hospitalization time. Currently in Lithuania there are no standards for burned patient nutrition. More attention is given to strategy of surgical strategy and techniques, as well as antibiotic therapy. This article is the review of the different aspects of artificial nutrition of burned patient: indications, modes of nutrition, mixtures and terms of nutritional support. PMID- 14764977 TI - [Direct and indirect diabetes costs in the world]. AB - Diabetes is becoming one of the major public health problems because a great proportion of the healthcare expenditure has been spent on the treatment of its associated morbidity and mortality. Diabetes is also a major cause of premature mortality, stroke, cardiovascular disease, peripheral vascular disease, congenital malformations as well as long- and short-term disability. In addition, persons with diabetic complications have a lower quality of life compared with persons without diabetes. The goal of this paper is to review the studies on the costs of diabetes, to identify the strengths and limitations of currently available diabetes cost studies, and to identify future research areas that will help us to better understand the economic burden of diabetes. The economic burden of diabetes mellitus is enormous in the world. Cost or illness estimates are often cited as an important element in the choices made regarding diabetes care and management. Studying these economic aspects presents several challenges, such as collecting the appropriate epidemiological and cost data, determining the diabetes attributable factors for premature morbidity and mortality, and determining methods to account for premature morality, disability, and reduced quality of life. The cost to care for diabetes puts a tremendous burden on both the patient and the payer. The direct cost of diabetes increased from 1.7 billion US dollars in 1969 to 44.4 billion US dollars in 1997. Several studies over the years have found that indirect costs related to diabetes are higher than direct. Indirect costs during 28 years increased 33 times, from 1.6 billion US dollars in 1969 to 54.1 billion US dollars in 1997. The expenses of one diabetic patient highly vary in different countries: from 13 US dollars in Bangladesh to 11,157 US dollars in USA per one year. Most of diabetes expenditure is used to pay for inpatient services (60-85%); the biggest part of it is incurred because of late diabetes complications (70%). Diabetes accounted to 3-12% of total healthcare expenditure in different countries. In this era of limited resources and escalating costs, it is critical to have an understanding of the economics of diabetes in order to develop and implement sound public health and prevention policies. PMID- 14764976 TI - Possible association between cell membrane band 3 impairment function and renal tubular acidosis (liver diseases, malignancies and adverse drug reactions). AB - Renal tubular acidosis (RTA) more frequently develops in case of chronic diseases of inflammatory-immunological origin. RTA is well known to be associated with chronic liver disease (CLD), with nephrolithiasis, common cases of RTA occur among cancer patients. Abnormalities in the expression or function of band 3 in cell membrane may play a role in the pathogenesis of RTA. Cl-/HCO3- anion exchanger (AE2) is an isoform of band 3 protein, which is expressed in cell membranes of organs such as liver cells and kidney endothelium. There are reports on downregulated AE2 immunoreactivity in the liver of patients with chronic liver diseases and in the kidney tubular tissue of patients with RTA. The proteolytic damage of cell membrane band 3 in tissues could be related to inflammatory immunological processes. Another important factor able to disturb the band 3 function is medicinal products used in the treatment of certain pathologies. The active substance of a drug itself may have a direct effect on this protein or trigger a pathological process. In such cases ADR can take place and may be evaluated as such. Acid-base disturbances, notably metabolic acidosis, are a serious complication of drug treatment. Reduced AE2 expression or its changed activity (congenital or acquired) could be related with alterations of intracellular pH. This could lead to antigenic changes and autoimmunity. The derangement of band 3 function in organ cell membrane could act as a factor which creates an "acidotic environment" for organ cells. Such circumstances could be the reason for unsuccessful treatment or determine resistance of tumor treatment. The understanding of the mechanisms of RTA development, early diagnostics, and knowledge of the drugs that can cause RTA, are of particular practical significance. PMID- 14764978 TI - [Risk factors affecting hospital resource use in cardiac surgery]. AB - Demographic changes in the society, favorable changes in disease patterns within population cause higher morbidity and higher mortality from cardiovascular diseases and increase need for hospital resource use. It is very important to identify the risk factors, which are responsible for higher health care costs. The objective of this article is to identify risk factors, which are responsible for higher hospital resource use. METHODS: Search in web and meta analysis of the electronic-medical articles within journals under review were performed. RESULTS: The main risk factors for higher hospital resource utilization were identified: longer length of stay in the hospital, type of operation, postoperative mortality, postoperative complications and age. CONCLUSIONS: The main factors affecting higher hospital resource utilization are following: longer duration of stay in the hospital, advanced type of operation, postoperative mortality, postoperative complications and advanced age. Hospital resource utilization depends on patient anamnesis and quality of care in institution. PMID- 14764979 TI - Blood serum apolipoproteins B and A-I in females suffering from rheumatic heart valve disease. AB - OBJECTIVE: With the aim to check whether the atherogenic factors are involved in the mechanisms of valve fibrosis, we have studied the blood serum concentrations of apolipoproteins (apo) A-I and B concentration in patients suffering from rheumatic heart valve fibrosis. METHOD: The quantities of apoA-I and apoB in the blood serum were tested by the ELISA method. Concentration of apoA-I and B in the blood serum was determined in rheumatic females with replacement of the damaged valves: after aortic valve operation (n=11; mean age 43.3+/-3.6 years) and after mitral valve operation (n=29; 41.3+/-4.1). The results obtained for rheumatic patients were compared with the data on age-matched healthy females (n=43; 39.5+/ 5.2 years). RESULTS: Significantly lower apoA-I level in the blood serum of all patients suffering from rheumatic heart valve disease was determined as compared with controls: in the pooled group of patients (1.02+/-0.22 vs 1.23+/-0.23 g/l, P<0.001), in women after aortic valve replacement (0.98+/-0.21 vs. 1.23+/-0.23 g/l, P<0.005), and in women after mitral valve surgery (1.03+/-0.23 g/l vs 1.23+/ 0.23 g/l, P<0.005). The apoB level in patients suffering from rheumatic heart valve disease did not differ from that of controls. The apoB/apoA-I ratio for patients with valve fibrotic damage was significantly higher as compared to controls in all groups (P<0.02). The increase of apoB/apoA-I ratio in patients with rheumatic valve fibrosis was caused by lower apoA-I levels in blood serum. CONCLUSION: The obtained results indicate that decreased apoA-I levels in blood serum can be indicative of valve fibrosis in rheumatic heart valve disease patients. PMID- 14764980 TI - [Transcutaneous electrical nerve stimulation in treatment of rheumatoid arthritis patients]. AB - The aim of the study was pain management in rheumatoid arthritis patients (n=10) by applying the method of transcutaneous electrical nerve stimulation. The 10 point visual analogue scale was used before stimulation and following it. The total number of pain measurement tests in the joints was 180. The estimated average pain before stimulation was evaluated by 6.56+/-0.17 points and following it by 5.88+/-0.18 points. The average pain decrease after stimulation (Pt<0.05) was 10.4 %, compared with the control level. PMID- 14764981 TI - [Clinical forms of new cases of tuberculosis at Kaunas Romainiai Tuberculosis Hospital in 1998-2001]. AB - Totally 1427 patients with tuberculosis were investigated in Kaunas Romainiai Tuberculosis Hospital. All patients belonged to the first category (new cases positive for mycobacterium tuberculosis, or severe tuberculosis). Infiltrated and disseminated tuberculosis were the most frequent clinical forms. The frequency of infiltrated tuberculosis ranged from 57.36 to 68.8%, and disseminated tuberculosis ranged from 18.52 to 30.36%. The most frequent complications were bleeding from the lungs and chronic cor pulmonale. The resistance to isoniazid, rifampicin and streptomycin was investigated. Multi-drug resistant tuberculosis did not exceed 1%. We suggest that so few cases of multi-drug resistant tuberculosis are due to implementation of WHO treatment standards. PMID- 14764982 TI - [Preoperative radiation with chemotherapy for rectal cancer: its impact on downstaging of disease and the role of endorectal ultrasound]. AB - OBJECTIVE: Preoperative adjuvant radiation combined with chemotherapy is a recent development in the management of patients with rectal cancer invading perirectal tissue and regional lymph nodes. This study was performed to assess the impact of preoperative adjuvant therapy in patients judged by endorectal ultrasound to have extramural invasion of rectal cancer and/or regional lymph node involvement on tumor regression in bowel wall T and lymph nodes N. The predictive value of ultrasound in staging wall penetration and lymph node involvement after preoperative adjuvant therapy was also assessed. MATERIALS AND METHODS: Fifty-one patients were selected by ultrasound to have preoperative irradiation (40-50 Gy over 5-6 weeks). In 29 patients this was combined with 5-fluorouracil chemotherapy. Assessments of ultrasound were compared with pathologic findings in the resected specimen in all patients. RESULTS: Partial downstaging was seen in 37 (72.5%) patients with wall invasion T and in five (9.8%) of 51 patients with lymph node involvement N. Complete downstaging was achieved in one (2.0%) patient with wall invasion T and in 20 (39.2%) of 51 patients with lymph node involvement N. Positive predictive values of ultrasound after irradiation were 47 (92.2%) and 45 (82.2%) for wall penetration and lymph node status, respectively. Negative predictive values of ultrasound after irradiation were rare 3.9% and 5.9%, respectively. CONCLUSIONS: In the majority of patients with rectal cancer invading perirectal tissues or lymph nodes, lesions downstages by preoperative chemo radiotherapy. Endorectal ultrasound examination before and after chemo radiotherapy for rectal cancer is one of the most recommended in staging rectal cancer. PMID- 14764983 TI - [Ischemic heart disease in women: prognostic value of ventricular repolarization variables]. AB - The aim of the study was to evaluate changes of ventricular repolarization variables during the exercise test in women with coronary heart disease, to determine variables, which can best predict significant coronary artery stenoses, and to improve noninvasive diagnostics of coronary heart disease. Ninety women with unstable angina pectoris undergone coronary artery angiography and exercise test on 4th-6th day of hospitalization at Kaunas University of Medicine Hospital. There was no difference in ventricular repolarization variables (JT interval, JT dispersion, JT and ST product) in women without coronary artery stenoses and those with one or two coronary artery stenoses. Sensitivity (87%), specificity (60%), positive predictive (43%), negative predictive (93%) and diagnostic value (67%) of stress test variable, which had no correlation with heart rate -ST and JT product- was greater comparing with ST depression greater than 0.1 mV (values respectively 61%, 49%, 29%, 79%, 52%). JT dispersion at the peak of exercise greater than 33 ms had the best diagnostic value (77%). The most accurate predictors of three-vessel disease are JT dispersion, ST and JT product at the peak of exercise and the fact of previous MI: JT dispersion at the peak of exercise more than 33 ms significantly increases three-vessel disease odds ratio 7.95 times. ST and JT product greater than 22,38 mV x ms significantly increases three-vessel disease odds ratio 13.9 times. The fact of previous MI significantly increases three-vessel disease odds ratio 6.85 times. PMID- 14764984 TI - [Giant euthyreotic retrosternal struma complicated by compression and thrombosis of brachiocephalic, subclavia, jugular and axillary veins, edema and necrosis of brain]. AB - The case of giant euthyreotic retro-sternal struma complicated by compression and thrombosis of brachiocephalic, subclavia, jugular and axillary veins, edema and necrosis of brain is presented in the article. 64-year-old man was admitted to the hospital due to ischemic stroke. Retrosternal struma was suspected according to clinical symptoms of compression of superior vena cava. Due to progression of neurologic symptoms the patient died. During autopsy giant euthyreotic retrosternal struma complicated by compression and thrombosis of brachiocephalic, subclavia, jugular and axillary veins, edema and necrosis of brain was found. Timely established diagnosis and effective surgical treatment of retrosternal struma could prevent such dangerous complications as edema and necrosis of the brain. PMID- 14764985 TI - [The survey of Lithuanian physicians and medical residents regarding possible migration to the European Union]. AB - The aim of this survey was to evaluate intentions of Lithuanian physicians and medical residents to work in the European Union (EU) and other countries. The survey was performed in 2002 in the framework of the project "Health human resource development and planning in Lithuania", sponsored by Open Society Fund, Lithuania. Using the questionnaire developed by the Ministry of Labor and Solidarity of France, 242 medical residents and 497 physicians were surveyed. Survey results indicated that 60.7% of medical residents and 26.0% of physicians intended to leave for the EU or other countries. The survey also showed that the first-choice countries were United Kingdom, Germany and the Nordic countries. Almost 15.0% of medical residents and 5.0% of physicians planed to leave for the EU on permanent basis. The largest part of those who intended to leave for the EU, planned to go there after accession. It was a definitive decision of 2.5% of medical residents and 3.8% of physicians. The major reasons for leaving were higher salary, better professional possibilities and better quality of life. For medical residents a previous visit abroad for professional reasons increased the risk of working abroad significantly (OR-3.29, 95% CI 1.73-6.27). In the case of physicians, age was the factor that significantly decreased the risk (OR-0.94, 95% CI 0.91-0.96); however having friends abroad increased the risk by more than three times (OR-3.22, 95% CI 1.91-5.42). In order to observe the possible migration to the EU or other countries, the survey should be repeated every three four years. PMID- 14764986 TI - [Trends in out-of-hospital mortality from ischemic heart disease in Kaunas middle aged population during 1983-1998]. AB - The aim of the study was to evaluate trends in out-of-hospital mortality from ischemic heart disease (IHD) in Kaunas population aged 25-64 years during 1983 1998, according to the ischemic heart disease register data. MATERIALS AND METHODS: The primary source of data was Kaunas population-based ischemic heart disease register. The main sources of information for registration of ischemic heart disease events were hospital discharge records, patient follow-up records of out-of-patient departments, death certificates, necropsy and medico-legal records. The data were collected according to the requirements of the WHO program MONICA. Only deaths from IHD that occurred out-of-hospital were analyzed in this study. The age-standardized rates were calculated by the direct method and the world standard population was used as the standard. Trends in rates were analyzed using the method of linear regression on logarithms of the age-standardized annual mortality rates. RESULTS: During 1983-1998, out-of-hospital IHD mortality was 137.9/100000 among Kaunas men aged 25-64 years meanwhile among women the corresponding rate was 7 times lower (20.2/100000). During 1983-1994, out-of hospital mortality from IHD was increasing and during 1995 to 1998 was decreasing (by 19.8% per year; p=0.03) among men. Among women, out-of-hospital mortality rates were increasing during 1983-1990, were rather stable during 1991-1994, and tended to decrease by 27.5% per year (p=0.09) during 1995-1998. When analyzing trends in rates by 10-year age groups, statistically significant changes in out of-hospital mortality from IHD were detected just among those aged 55-64 years. Among men, out-of-hospital mortality from IHD tended to increase during 1983 1994, and was decreasing statistically significantly (by 22.9% per year; p=0.007) during 1995-1998. Among women, the corresponding rates were increasing by 26.7% per year (p=0.03) during 1983-1986, and tended to decrease by 40.1% per year, (p=0.09) during 1995-1998. During 1983-1998, out-of-hospital deaths from IHD accounted on the average for 76.5% among all deaths from IHD among men and for 66.9% among women. During 1983-1998, the proportion of out-of-hospital deaths among all deaths from IHD remained without statistically significant changes among both men and women. CONCLUSION: During 1983-1994, out-of-hospital mortality rates of Kaunas middle-aged men increased and during 1995-1998--significantly decreased. Out-of-hospital mortality rates of Kaunas women increased during 1983 1990, but in 1995-1998--tended to decline. Out-of-hospital deaths from IHD accounted on the average for 76.5% of all deaths from IHD among men and for 66.9% of women deaths, and biggest part of these deaths were among younger persons. PMID- 14764987 TI - Master of Pharmacy Wilhelm Grining and his pharmaceutical laboratory. AB - Very little is known about the history of pharmaceutical industry in Latvia, especially about the history of small pharmaceutical enterprises in Riga at the turn of 19-20th centuries. The laboratory of Wilhelm Grining, Master of Pharmacy, (established in 1899) became well known because of its product Liquor Ferri albuminati Grining. The liquor was used for treatment of anemia and became very popular in the whole Russian empire. The owner of the laboratory Wilhelm Grining was a very erudite man and also a good businessman. He was a son of a pharmacist and has been acquainted with apothecary's practice and specificity of the work of the owner of a pharmacy since his childhood. Wilhelm Grining was particularly interested in proteins and started to carry out scientific research both in the fields of chemistry and pharmacy. He participated with his reports in the congresses of Russian Pharmacy Society and published his works in the journal of Russian Pharmacy. Besides all this W. Grining is believed to be the author of the concoction of curative herbs "Trejos devynerios". Biography of W. Grining, as well as the products produced by his pharmaceutical laboratory, was explored using materials from the archive of Latvian State history and 19th century's German literature of pharmaceutical history. PMID- 14764988 TI - [Intravenous catheters and nosocomial infection]. AB - Peripheral, especially central venous catheters, are used with increasing frequency in the intensive care unit and in general medical wards to administer intravenous fluids and blood products, drugs, parenteral nutrition, and to monitor hemodynamic status. Catheter infection is associated with increased morbidity, mortality, and duration of hospital stay. Risk factors in the development of catheter colonization and bloodstream infections include patient factors (increased risk associated with malignancy, neutropenia, and shock) and treatment-related factors (increased risk associated with total parenteral nutrition, intensive care unit admission for any reason, and endotracheal intubation). In this review article terms and definitions of catheter-related infections, pathophysiology and epidemiology of "catheter sepsis", factors determining risk of infection, catheter types and materials, insertion procedure, choice of insertion site, indwelling time, dressing and care of the insertion site, various preventive strategies and future developments, special situations and procedures, and treatment are discussed. Reducing catheter infections rates requires a multiple-strategy approach. Therefore, intensive care units and other locations where catheters are used should implement strict guidelines and protocols for catheter insertion, care, and maintenance. PMID- 14764989 TI - The functional interactions between the p53 and MAPK signaling pathways. AB - The p53 tumor suppressor protein exerts its growth inhibitory activity by activating and interacting with diverse signaling pathways. As a downstream target, p53 protein is phosphorylated and activated by a number of protein kinases in response to stressful stimuli. As an upstream activator, activated p53 acts as a transcription factor to induce and/or suppress a number of genes whose expression leads to the activation of diverse signaling pathways. p53 protein can also interact with a number of proteins, resulting in an increase or decrease in p53 activity itself. The activation of p53 leads to many outcomes in cells, including cell cycle arrest and apoptosis. It has become clear that the p53 protein can functionally interact with the mitogen-activated protein kinase (MAPK) pathways, including the stress-activated protein kinase [SAPK/c-Jun N terminal protein kinase (JNK)], the p38 mitogen-activated protein kinase (MAPK), and the extracellular signal related kinase (ERK). Upon exposure to stressful stimuli, MAP kinases phosphorylate and activate p53, leading to p53-mediated cellular responses. Recent studies have suggested a role of p53 as an upstream activator to regulate MAPK signaling via the transcriptional activation of members of the dual specificity phosphatase family. Because both the p53 and MAPK signaling pathways are altered in the majority of human tumors, understanding their functional interaction may provide new insights into the deregulated cell proliferation and survival that is characteristic of cancer. PMID- 14764990 TI - Upregulation of the androgen receptor during prostate cancer progression. AB - The androgen receptor is one of the central factors in mediating prostate cancer progression and is an important target for treatment. Several possible mechanisms have been put forth as to how it promotes this process. In the January 2004 issue of Nature Medicine, Chen et al. report that the androgen receptor is consistently upregulated as hormone sensitive prostate cancer changes to a hormone refractory state. They also show that this change is intricately involved during the development of resistance to androgen ablation therapy. PMID- 14764991 TI - Liposomal mediated transfer of ErbB2 antisense DNA: coming of age in the war against cancer. PMID- 14764992 TI - Glycobiology and cancer: meeting summary and future diections. PMID- 14764993 TI - Targeting aberrant signal transduction pathways in lung cancer. AB - Lung cancer is the deadliest form of cancer in the world and is most commonly associated with smoking. Current treatment strategies are largely ineffective due to advanced stage at diagnosis and the inherent therapeutic resistance of lung cancer cells. To improve patient outcomes, many studies have been designed to identify molecular alterations in lung cancer in order to develop new therapeutic strategies. Molecular alterations in lung cancer include genetic changes, epigenetic changes, and changes in the expression or activity of kinases that comprise signaling pathways within cells. Signaling pathways are attractive targets for lung cancer therapy because activation of signaling pathways contributes to tumor growth and therapeutic resistance, and constitutively active signaling commonly occurs in lung cancer. This review will discuss signaling pathways that are relevant to lung cancer. We will discuss specific signaling aberrations found in lung cancers, review the status of signaling inhibitors being developed for lung cancer, identify emerging targets, and provide recommendations for the development of agents designed to inhibit signal transduction. PMID- 14764994 TI - Gene expression profiling of human cutaneous melanoma: are we there yet? PMID- 14764995 TI - Targeting Sonic Hedgehog: a new way to mow down pancreatic cancer? AB - Despite continuing development of new therapies, the prognosis for patients with pancreatic cancer remains extremely poor. In part, this may relate to molecular abnormalities that stimulate pancreatic tumorigenesis and also contribute to reduced sensitivity to standard treatments such as chemotherapy and radiotherapy. Two recent reports in Nature suggest that Sonic Hedgehog (Shh) overexpression may contribute to pancreatic tumorigenesis and that cyclopamine, a specific inhibitor of Shh signaling, can reduce pancreatic cancer cell growth and viability. This discovery is exciting and suggests that targeting Shh signaling may be an effective novel approach to therapy in patients with this devastating disease. PMID- 14764997 TI - Real-life-type problem solving in Tourette syndrome. AB - OBJECTIVE: The main objective of the study was to examine social problem solving in real-life-type situations in Tourette syndrome (TS). BACKGROUND: Studies of cognitive functioning in TS have usually focused on nonsocial, abstract tasks, with mixed findings as to whether there is evidence of impairment in executive functions in those without comorbid disorders. The current study focuses primarily on social functioning, using a problem-solving task known to be sensitive to frontal lobe lesions. METHODS: TS participants without comorbid diagnoses were compared with matched healthy control participants on a problem solving task, using a range of interpersonal problem scenarios presented on video. A set of more abstract executive tests was also included. RESULTS: Participants with TS were found to perform below a matched control group on the problem-solving task both in generating a range of potential problem solutions, and in selecting appropriate final solutions. They also performed more poorly on aspects of executive function. CONCLUSIONS: This study provides evidence of difficulties in both social and nonsocial aspects of functioning in TS. The implications of the findings for our understanding of TS and problem solving are discussed. PMID- 14764998 TI - False recognition in Alzheimer disease: evidence from categorized pictures. AB - OBJECTIVES: To better understand memory distortions and false recognition in patients with Alzheimer disease (AD), using a paradigm of categorized color photographs. BACKGROUND: Previous research has found that patients with AD and older adults showed similar levels of uncorrected false recognition of semantic associates and of perceptually related novel objects. In contrast to these results, using a paradigm in which semantically related words were accompanied by black and white line drawings, it was found that patients with AD showed a trend toward higher levels of uncorrected false recognition compared with older adults. METHODS: To explore this trend, 24 patients with AD and 24 older adults matched for age, education, and gender were examined using a false recognition paradigm consisting of categorized color photographs (e.g., flowers, motorcycles, cats). RESULTS: Compared with older adults, patients with AD showed higher levels of uncorrected false recognition, but lower levels of corrected false recognition and lower levels of item-specific recollection. CONCLUSIONS: The authors suggest that these results may be attributable to the poor ability of patients with AD to acquire both gist and item-specific information as well as these patients' inherent frontal lobe dysfunction leading to difficulty inhibiting responses on the basis of familiarity alone. PMID- 14764999 TI - Deciphering components of impaired working memory in multiple sclerosis. AB - OBJECTIVE: To identify working memory (WM) impairment by examining components of auditory working memory with the same sample of individuals with multiple sclerosis (MS). BACKGROUND: Although individuals with MS have consistently demonstrated WM impairments, the specific components involved (i.e., central executive versus storage/maintenance) remain unclear. METHOD: Individuals with MS with and without cognitive impairment and healthy controls were administered a task primarily assessing storage and rehearsal/maintenance (auditory n-back) and a task with a significant central executive component (Paced Auditory Serial Addition Task). RESULTS: Results indicate when the phonological loop is minimally challenged, maintenance of information is generally efficient. However, the addition of a central executive component to processing appears to be the key factor in differentiating individuals with MS with cognitive impairment from those without cognitive impairment and healthy controls. CONCLUSIONS: Results indicated that the primary WM impairment in MS is within the central executive rather than the phonological loop. PMID- 14765000 TI - Clinical correlates of cognitive decline in vascular dementia. AB - OBJECTIVE: To determine whether demographic data, dementia severity, functional status, whole brain volume (WBV), or subcortical hyperintensity volume (SH) predict subsequent cognitive decline in vascular dementia (VaD). BACKGROUND: The identification of variables that accurately predict progressive cognitive decline in dementia has important clinical implications. METHODS: A cohort of 30 patients with VaD completed neurologic and neuropsychologic examinations and magnetic resonance imaging of the brain at baseline and again after 12 months. All participants met clinical and research criteria for VaD according to standard guidelines. Change scores were computed for measures of verbal fluency, verbal learning, and visual learning. Potential correlates of cognitive change included age, education, score on the Hachinski scale, WBV, SH, and functional ability. RESULTS: As a group, lower WBV and lower Hachinski score correlated with decline in verbal fluency and visual learning, whereas lower Hachinski score correlated with decline in verbal learning. However, when subdivided by disease type, this pattern held only for individuals with evidence of a cortical stroke at baseline. No clinical variables correlated with cognitive decline among individuals without a cortical infarction. CONCLUSIONS: Assessment of cognitive decline in VaD should be guided by dementia subtype, with particular attention directed at severity of cerebral atrophy rather than classic symptoms of infarction. PMID- 14765001 TI - Brain damage and addictive behavior: a neuropsychological and electroencephalogram investigation with pathologic gamblers. AB - BACKGROUND: Gambling is a form of nonsubstance addiction classified as an impulse control disorder. Pathologic gamblers are considered healthy with respect to their cognitive status. Lesions of the frontolimbic systems, mostly of the right hemisphere, are associated with addictive behavior. Because gamblers are not regarded as "brain-lesioned" and gambling is nontoxic, gambling is a model to test whether addicted "healthy" people are relatively impaired in frontolimbic neuropsychological functions. METHODS: Twenty-one nonsubstance dependent gamblers and nineteen healthy subjects underwent a behavioral neurologic interview centered on incidence, origin, and symptoms of possible brain damage, a neuropsychological examination, and an electroencephalogram. RESULTS: Seventeen gamblers (81%) had a positive medical history for brain damage (mainly traumatic head injury, pre- or perinatal complications). The gamblers, compared with the controls, were significantly more impaired in concentration, memory, and executive functions, and evidenced a higher prevalence of non-right-handedness (43%) and, non-left-hemisphere language dominance (52%). Electroencephalogram (EEG) revealed dysfunctional activity in 65% of the gamblers, compared with 26% of controls. CONCLUSIONS: This study shows that the "healthy" gamblers are indeed brain-damaged. Compared with a matched control population, pathologic gamblers evidenced more brain injuries, more fronto-temporo-limbic neuropsychological dysfunctions and more EEG abnormalities. The authors thus conjecture that addictive gambling may be a consequence of brain damage, especially of the frontolimbic systems, a finding that may well have medicolegal consequences. PMID- 14765002 TI - Assessing the behavioral consequences of multiple sclerosis: an application of the Frontal Systems Behavior Scale (FrSBe). AB - OBJECTIVE: The current study examines the utility of the Frontal Systems Behavior Scale (FrSBe) to document behavioral changes in people with multiple sclerosis (MS). BACKGROUND: MS results in widespread central nervous system lesions that have a propensity for affecting frontal and parietal brain regions. As a result, behavioral symptoms may present, including disorders of executive functioning, apathy, and disinhibition. METHOD: Twenty-six people with MS and 15 healthy controls (HC) underwent neuropsychological testing and completed the Frontal Systems Behavior Scale (FrSBe), and measures of emotional functioning. RESULTS: Behavioral ratings for the MS group prior to illness were equivalent to those of the HC group. After illness however, a clinically and statistically significant increase was noted on two of three FrSBe scales, the apathy scale and the executive dysfunction scale. Relationships were also noted between the FrSBe family and self-report forms after illness and measures of neuropsychological performance, particularly information processing, working memory, and executive control. In contrast, physical disease progression was correlated with family ratings of behavior, but not self-ratings. CONCLUSIONS: The FrSBe appears to be a sensitive measure of behavioral change in people with MS. Self-ratings and family ratings of behavioral changes are related to measures of neuropsychological performance. However, physical symptoms of MS are related only to the family ratings of behavior. PMID- 14765003 TI - Orienting attention in obsessive-compulsive disorder. AB - OBJECTIVES AND BACKGROUND: Obsessive-compulsive disorder (OCD) is characterized by recurrent obsessions and/or compulsions that affect daily functioning. This study aimed to ascertain whether OCD patients have problems in orienting attention, via the use of a vibrotactile choice reaction time task. METHODS: Participants were required to depress a button in response to a vibration delivered to the index finger of either the right or left hand. Prior to the stimulus, one of three precues was administered: valid (precue delivered to the same hand as stimulus), invalid (precue delivered to opposite hand to stimulus), or neutral (precue delivered simultaneously to both hands). RESULTS: Obsessive compulsive disorder participants were overall slower and less accurate than controls. Contrary to our predictions, OCD participants did not show increased costs or benefits. However, unlike controls, patients did not show the normal pattern of faster reaction times on neutral compared with invalid precues, which may reflect a problem in inhibiting irrelevant information. CONCLUSIONS: Both patients and controls may benefit from the provision of directed attention in tactile tasks when difficulty levels are maximal. OCD patients may have problems with inhibitory control. PMID- 14765004 TI - Quantitative signal intensity measures on magnetic resonance imaging in attention deficit hyperactivity disorder. AB - OBJECTIVE: The aim of this study was to investigate possible abnormalities of cerebral myelination in subjects with attention-deficit hyperactivity disorder (ADHD). BACKGROUND: Anatomic and functional neuroimaging studies of subjects with ADHD demonstrated a right frontostriatal deficit and abnormal cerebral asymmetries. Some also reported white matter abnormalities, such as smaller white matter volumes in the right anterior-superior frontal region, and the smaller bilateral retrocallosal region. Smaller volumes in specific areas of the corpus callosum have also been reported. We hypothesized that white matter signal intensities may also show differences indicating abnormal cerebral myelination. METHOD: We analyzed T2-weighted magnetic resonance images of 11 adolescents with ADHD and 20 controls. Regions of interest were set in both the white and gray matter in frontal and parieto-occipital associative regions. RESULTS: The ADHD group showed a higher signal intensity ratio, probably reflecting a higher degree of myelination. Significant interhemispheric differences emerged only in the posterior region in the ADHD group. CONCLUSIONS: The higher degree of myelination in the right frontal region of ADHD may be due to a compensatory mechanism for the right frontostriatal dysfunction. PMID- 14765005 TI - Nonlinear effects in behavioral changes in Huntington disease. PMID- 14765007 TI - Variant Creutzfeldt-Jakob disease: need for mental health and palliative care team collaboration. AB - People with a dementia syndrome and a superimposed terminal illness are increasingly being referred to palliative care services. Creutzfeldt-Jakob disease (CJD) is a dementia syndrome in the early stages of which people experience a variety of psychological symptoms that may lead to them being admitted to psychiatric services. People with CJD have died in psychiatric units. There is clearly a need for collaboration between mental health and palliative care services in providing care for people with CJD and other dementia syndromes. An inductive qualitative study of one case of variant CJD (vCJD) was carried out to explore issues that were raised in providing care for a young person with the disease who had died in a hospice unit. The study, which was a pilot for a larger study, highlights some of the complexities of providing care for people with an end-stage dementia and identifies the need for the development of education initiatives to increase knowledge and understanding of the end-of-life needs of people with CJD and other dementia syndromes, and their families. PMID- 14765008 TI - District nurse involvement in providing palliative care to older people in residential care homes. AB - Although less than 15% of older people in care homes die of a terminal disease such as cancer, many more die following a period of slow deterioration. In the UK, residents of care homes receive their nursing care from primary healthcare providers. This article reports on a study that describes how district nurses and care home managers define their responsibilities when caring for residents who are dying. The findings presented are from a detailed survey of 89 district nurse team leaders and 96 care home managers. District nurses were the most frequent health professionals visiting care homes, but less than half reported involvement in palliative care. The differing priorities and perceptions of participants regarding what constituted nursing and personal care influenced older people's access to palliative care services. Furthermore, the organization of care and the stability of the care home workforce affected participants' ability to establish ongoing supportive working relationships. It is argued that unless there is an organizational review of current practice, older people will continue to have unequal access to generalist and specialist palliative care. PMID- 14765010 TI - A Greek perspective on concepts of death and expression of grief, with implications for practice. AB - Death has been conceptualised in different ways by different cultures and civilizations. It is increasingly entering into the public consciousness and society is now more ready to discuss and lessen the fear of dying and grief than it has been in the past few decades. In Greece, by Classical times there was an increase in burial rituals and commemorative practices compared to earlier periods. When Christianity was introduced into Greece it attempted to change the way the dead were mourned, preaching immortality of the soul and resurrection of the dead. Nevertheless, the way people grieve and bury their dead in Greece has not changed greatly since before the introduction of Christianity, except for the difficulty experienced in witnessing burial procedures observed in the large cities. Burial and bereavement traditions were introduced to help Greeks cope with death and bereavement. In Greece today beliefs about grief and death are based both on the ancient and the Christian Orthodox traditions. Healthcare professionals need to develop cultural competence to improve nursing and future health care. If care is culturally informed and tailored its quality is improved. PMID- 14765009 TI - Impact of the Powys Macmillan GP clinical facilitator project: views of health care professionals. AB - The UK charity, Macmillan Cancer Relief, commissioned a 3-year pilot project employing 12 GP clinical facilitators (GPCFs). The aim was to raise the standard of generalist palliative care, provide extended clinical palliative care and provide a coordinated framework for commissioning specialist palliative and cancer care in Powys, rural Wales. As part of the comprehensive evaluation, surveys of GPCFs, GPs, district nurses and community hospital nurses were undertaken in order to record changes in palliative care activity, specialist palliative care services and training needs. Services providing 24-hour nursing and social services were perceived as in need of development. Referrals to Macmillan nurses increased by 40% and GPs reported that time spent on palliative care increased, on average, from a quarter of a day to half a day per week, although district nurses reported a reduction in palliative care activity. The majority of nurses thought that the GPCF's contribution was important. The use of local palliative care guidelines increased significantly among district nurses by the end of the project. PMID- 14765011 TI - Models of lymphoedema service provision across Europe: sharing good practice. PMID- 14765012 TI - Proactive palliative care choices for haematology day unit patients. AB - End stage haematology patients may require transfusion of blood, or blood products, regularly throughout the week. One regional haematology day unit sister identified difficulties that these hospital services dependent patients were experiencing in establishing contact with local community specialist palliative care services. As a consequence, some patients were unable to access specialist palliative care services in the community when they were too unwell to attend the day unit, resulting in acute admission to hospital for terminal care. Discussion of this issue with nursing colleagues from the hospital and community palliative care teams led to the development of an alternative model of palliative care delivery for these patients. The new model increased patient choice. Evaluation of this model of care demonstrated an increase in planned hospice/community deaths, avoiding unnecessary emergency hospital admissions in the last days of life. All regional patients accessed some palliative care services/treatment locally and there was opportunity for patient-initiated palliative care review. PMID- 14765013 TI - Prostate cancer update. AB - Prostate cancer continues to be the most commonly diagnosed malignancy in men of the western world. Its diagnosis, evaluation, and treatment therapies are constantly evolving. This paper summarizes and clarifies the most up-to-date information on prostate cancer prevention and treatment. PMID- 14765014 TI - Prostate biopsy. AB - Transrectal ultrasound guided (TRUS) prostate biopsies is the standard method in the diagnosis of prostate cancer. The use of prostate specific antigen (PSA) and digital rectal examination for prostate cancer screening has led to a dramatic increase in the number of TRUS guided biopsies. Frequently urologists are faced with the dilemma of treating a patient with a high suspicion of prostate cancer, but an initial set of negative biopsies. In this review we focus on the current knowledge of prostate biopsies, the indication to perform a biopsy, the impact of prostate volume in the number of cores taken, the technique of an initial and repeat biopsies and when to stop. PMID- 14765015 TI - The evolving role of androgen deprivation therapy in the management of prostate cancer. AB - Androgen deprivation therapy (ADT) plays a central role in the management of prostate cancer. ADT is the mainstay of treatment for metastatic disease; the most common method is gonadal suppression via luteinizing hormone release hormone (LH) agonists, with or without antiandrogens. Antiandrogen monotherapy remains investigational, as is the appropriate role of 5alphareductase inhibition for prostate cancer. Intermittent ADT offers the promise of improved quality of life and reduced cost without a decrease found to date in oncologic efficacy. A growing menu of options exists for secondary androgen deprivation after disease progression on primary therapy: these include high-dose antiandrogens, estrogens, and adrenal androgen suppressants. ADT is being used with increasing frequency as primary monotherapy in patients with localized disease, but only small, nonrandomized studies of highly selected patients have been reported to date. Neoadjuvant ADT (NADT) has been demonstrated in prospective, multi-institutional trials to improve outcomes for patients with high-risk or locally advanced disease undergoing external-beam radiotherapy. Trials for patients with lower risk, localized disease are still ongoing. Neoadjuvant therapy does not improve outcomes for patients with localized disease opting for radical prostatectomy (RP) and has not been well studied in association with brachytherapy. The side effects of ADT can be managed increasingly successfully; in particular, the introduction of zoledronate may reduce the impact of ADT-associated osteoporosis. Finally, contemporary practice pattern data suggest that use of ADT is increasing across patient risk groups, both in contexts where such therapy is well supported by current evidence and in others where it is not. PMID- 14765016 TI - Laparoscopic radical prostatectomy: a review of techniques and results worldwide. AB - Laparoscopic radical prostatectomy (LRP) is currently performed in multiple centers world-wide, with several different surgical approaches and techniques utilized. A comprehensive review of the published literature worldwide on laparoscopic radical prostatectomy was performed to outline the evolution of this technique, and to review the published surgical, oncological and functional results. A systematic review of peer reviewed articles concerning laparoscopic radical prostatectomy was obtained using Medline query. LRP is being performed in multiple centers worldwide, using a variety of surgical approaches and technologies. Analysis of perioperative parameters, including surgical blood loss, operative time, complications and convalescence, demonstrates a low morbidity and shows a clear trend in improvement with increased experience. The functional results, as recorded by postoperative urinary and sexual functions, appear encouraging. The reported positive surgical margin rates decrease with more recent series. Oncological results and cancer control rates as measured by PSA recurrence and disease-free intervals are difficult to ascertain in the immature series published to date. LRP has witnessed tremendous popularity and widespread implementation in specialized centers worldwide. LRP represents a technically demanding laparoscopic procedure with a difficult learning curve, but can be performed systematically with standard techniques. The advantages include shorter convalescence and markedly lower operative blood loss, with quicker removal of the urinary catheter. Long-term functional and oncologic results are not yet available. PMID- 14765017 TI - Disease recurrence after radical prostatectomy. Contemporary diagnostic and therapeutical strategies. AB - In this paper the authors provide an overview of contemporary diagnostic and therapeutic strategies in patients with disease recurrence after radical prostatectomy. Literature on disease progression after radical prostatectomy (RP) is reviewed and a selection of articles made. Key words used for the Medline research included: prostate cancer (PC), RP, disease recurrence, prostate specific antigen (PSA) progression and biochemical failure (BF). Within 10 years following RP for clinically localized PC, about 1/3 of patients will present disease recurrence. This is generally diagnosed by BF. The prognosis of these men may vary considerably. Differences in PSA kinetics (PSA doubling time, PSADT, onset of PSA rise) are useful for differentiating between local recurrence and distant disease. Indications and results are provided for different treatment strategies such as local radiation therapy, hormone therapy or watchful waiting. The present paper reviews the recent international literature. Diagnostic strategies and therapeutic manoeuvres are discussed. Prognostic factors as well as treatment indications are presented with the aim of applying an individual therapy. PMID- 14765018 TI - External beam radiotherapy as definitive treatment for prostate cancer. AB - External beam radiotherapy (EBRT) planning and delivery has dramatically changed in the last 2 decades allowing dose escalation to the prostate while maintaining the same risk of complications. EBRT plays a major role in the treatment of localized (T1-2) and locally advanced (T3-4) prostate cancer. In this review we will focus on some current issues associated with the "modern" use of EBRT such as what to expect from EBRT in terms of tumor control and side effects, what is the role of biopsy and surgery after EBRT, what is the role of androgen deprivation (AD), and how EBRT compares to brachytherapy. PMID- 14765019 TI - [Morphologic evaluation of adhesive/resin cement system and fiber post: a sem investigation]. AB - AIM: The purpose of the present paper is to carry out an SEM investigation into the adhesive cementation of quartz fibre posts in canals treated with endodontic cements containing or not containing eugenol. METHODS: Sixteen selected tooth roots were subdivided into 4 groups and prepared with the step-back technique. Group A, without canal filling and with a cemented post, was the control group; in groups B and C, the canals were closed with guttapercha, endomethasone C and eugenol, those of group D with guttapercha and AH Plus without eugenol. The canals were prepared so as to be able to take D.T. light-posts irrigated with water (group C with ethyl alcohol) mordanted with orthophosphoric acid after applying 2 layers of photopolymerised ONE-STEP; 2 layers of ONE-STEP were applied to the post. DUO-LINK cement was positioned in the canal after inserting the post, removing excess resin and photopolymerising. The samples were prepared for SEM observation. RESULTS: Group A: the apical portion shows the post absorbed into the cement and resin tags; group B: porosity is noted between cement and post, the hybrid layer does not present tissue penetration of the dentin resin; group C: bullae can be seen between cement and post surface and between hybrid layer and cement; group D: there is relative continuity between post surface, cementing resin, hybrid layer and underlying dentin. CONCLUSION: The study has shown that eugenol interferes with the formation of the hybrid layer, the pictures relative to the resinous cement appeared different depending on whether the canal was treated with cement containing or not containing eugenol. PMID- 14765020 TI - [Incisional biopsy in oral medicine: punch vs traditional procedure]. AB - AIM: In oral medicine the incisional biopsy, removing a specimen of tissue for microscopic examination, represents an important step in diagnosis and management. Up till now, many practitioners have used the traditional scalpel 15 and, recently, the scalpel punch, an easy and quick device with a circle lama, has been introduced. The aim of the present study was to compare clinical effects of the punch biopsy technique versus traditional method with scalpel 15. METHODS: This prospective case-control study investigated 41 patients [mean age 56.1, range (21-79); 16 males (39.0%), 25 females (61,0%)], consecutively referred to the Unit of Oral Medicine in Palermo (July-December 2002); in parallel groups, 18 subjects [mean age 58, range (36-79); 6 males (33.3%),12 females (66,6%)] were treated with punch scalpel (TEST group) and 23 [mean age 54.6, range (21-77); 10 males (43.5%), 13 females (56,5%)] with traditional scalpel (control group). A single operator carried out all biopsy procedures; a different single observer, in 3 different times, registered the following outcomes: 1) postoperative pain by mean a Visual Analogic Scale (VAS), 2) timing of wound-healing, 3) restitutio ad integrum. RESULTS: There was no statistically significant differences between the 2 techniques for the above parameters. CONCLUSION: Punch biopsy, an easy and quick technique, may be considered a valid procedure in oral medicine for the diagnosis and the follow-up of several lesions. PMID- 14765021 TI - [The cellular and molecular bases of "tooth framing"]. AB - Progress in molecular biology in recent years has enormously increased interest in tooth generation. The enamel knot has been discovered, in consequence. This is a transient structure acting as molecular signaling center, responsible for controlling cusp formation, stimulating growth of surrounding epithelium, and generating new knots or their disappearance through apoptosis. Both tooth development and enamel knots are regulated by a cascade of gene activity where Fgf4, Shh, BMP4, Lef1 and p21 are the prime movers of the processes. Homeobox genes (Msx, Dlx) are the orchestrators of the framing and a series of proteins (adhesion molecules, extracellular matrix components) are the executors of "tooth framing". An important concept has emerged from developmental biology through the identification of the basic mechanisms involved in tooth development: the molecular basis of structure framing shares common rules. Thus similar genetic programs are involved in body structure generation (limb bud, tooth, branching morphogenesis). A deeper understanding of developmental rules regulating tooth formation will make it possible in the near future: a) to modify in vivo homeobox gene expression and restore tooth generation hampered by tooth agenesia due to homeobox gene deregulation; b) to induce complete tooth formation, in case of tooth loss due to trauma or diseases, through implantation in the patient's oral cavity of a synthetic ball containing morphogens and growth factors to stimulate, in the right spatio-temporal sequence, the entire tooth genetic cascade. These concepts will certainly enforce cultural and practical interaction between biology and dentistry. PMID- 14765022 TI - [Burning mouth syndrome]. AB - Burning Mouth Syndrome (BMS) is a frequent disease characterized by a burning or painful sensation in the tongue and/or other oral sites without clinical mucosal abnormalities or lesions. The etiopathology is unknown although local, systemic and psychological factors have been connected with BMS. As this syndrome is a multifactorial disease, the diagnostic and therapeutic approach should be multidisciplinary. In this paper a review of the literature is presented and the most recent advancement on clinical, etiologic, diagnostic and therapeutic aspects of BMS are discussed. PMID- 14765023 TI - Dental anomalies associated with genetic factors. AB - The article reviews the association between dental anomalies and genetic diseases. Knowledge of these hereditary diseases will enable the dentist to make a more extensive analysis of the families in which such diseases occur, and above all to evaluate more precisely the dental aspects that such conditions favour. This will lead to better management of diseases linked to malocclusion, congenital absence of teeth and anomalous development of dental tissues. Hereditary diseases are caused either by a single mutant gene, inherited from 1 parent and acting as dominant gene, or from a pair of mutant genes, inherited from both parents and acting recessively. Autosomal-dominant and autosomal recessive inherited diseases are described, together with hereditary sex-linked conditions and those of multi-factorial origin. The purpose is to review such diseases so that the dentist can make an early diagnosis in clinical practice, check the predisposition toward such diseases and put in place preventive measures. PMID- 14765024 TI - [Actinomyces parotid infection after mandibular third molar extraction]. AB - Actinomycosis is currently an uncommonly diagnosed human disease. The disease is a chronic suppurative infection caused by micro-organism from the Actinomyces group, most often Israelii. A patient with cervicofacial actinomycosis generally reports a history of recent dental treatment which, usually, involves extraction of a mandibular molar. The common initial signs and symptoms of infection (such as sudden onset of cervicofacial pain, swelling, erythema, edema and suppuration) can be absent. In this case report a 29-year-old man presented a mass in his left parotid area, 1 week after mandibular molar extraction. Echography and CT scans revealed a parotid abscess area. The needle-biopsy of swelling revealed infection due to Actinomyces. Therapy was started with intravenous cefazolin (fl 1g X 2 in 100 s.s. i.v.) and methylprednisolone (25 mg tablet, 1/die) for 9 days; 14 days after treatment suspension the lesion reappeared with a fistula and a new therapy was given (ceftriaxone 1 g/die and gentamicin 80 mg/i.m. for 3 weeks). To prevent a relapse, the patient received cefalexin 1 gx2/die per os for 4 weeks. After a follow-up of 1 year, the patient was still asymptomatic. PMID- 14765025 TI - Florid cemento-osseous dysplasia. Report of a case in a white woman with both mandibular and maxillary involvement. AB - Florid cemento-osseous dysplasia (FLCOD) is a rare pathological entity whose classification is not yet completely defined although its non-neoplastic clinical and histologic behaviour is very clear. FLCOD is a self-limiting lesion scarcely delimited from the surrounding healthy tissue and whose surgical treatment therefore is not elective, unless inflammatory signs are present. A case of a white woman with both mandibular and maxillary involvement is presented; she was referred to us for partial impaction of her lower 3(rd) molars. Standard radiographic and CT images are examined and the patient management is discussed. PMID- 14765026 TI - Cancer of the head and neck: general principles of radiobiology, radiotherapy and chemotherapy. Radiation damage. AB - The overall incidence of tumours of the head and neck area versus tumours of other areas is 10%; in other words, 1 neoplastic lesion out of 10 affecting the human body is in this area. The relative frequency places malignant tumours of the larynx in 1(st) place with 25-30%. This data relates only to the areas of competence of the dentist and ENT specialist, and thus indicates that it is now becoming increasingly frequent for the dentist to be faced with treating a cancer patient. At the same time, it is not possible to consider as "dental cancer patients" only those patients whose disease involves an anatomic structure of the oral cavity as such. Pa-tients with tumours involving any of the anatomic structures of the head and neck area (pharynx, larynx, lip, oral cavity, pa ranasal sinuses, nasal fossae, salivary glands) must also be included. This article provides information on the epidemiology of tumours of the head and neck, the theoretical foundations of radiobiology, radiotherapy and chemotherapy, and possibilities for integration among the different possible treatment methods, i.e. radiotherapy, surgery, and chemotherapy. It looks at the most recent developments in these methods for patients with cancer of the head and neck, including the diagnostic protocol (clinical and instrumental). Lastly, irradiation damage to this anatomical area is analysed in terms of acute and delayed manifestations. PMID- 14765027 TI - Orofacial cleft in Southern Italy. AB - AIM: Cleft lip and palate or orofacial cleft (OFC) is one of the most common congenital malformations. The average incidence is around 1 every 1 000 live births. Different types of cleft lip and palate exist: cleft lip (CL), cleft lip and alveolus (CLA), cleft lip, alveolus and palate (CLP), and cleft palate only (CPO). Genetic studies on human samples have demonstrated that OFC has a heterogeneous genetic background and environmental factors also contribute to disclose this malformation. Because of the complex aetiology of OFC, studies on different and homogeneous populations can be useful in detecting environmental and genetic factors involved in the onset of this disease. The aim of the present study is to evaluate the relation between gender, type of cleft and affected side in a group of patients in Southern Italy. METHODS: Six hundred and fifty patients were enrolled in this retrospective study. They were operated at the Dental Clinic of the Second University of Naples in the period 1980-2002. Gender, type of cleft and affected side were analysed by means of the "Test for comparing two proportions". RESULTS: Among the analysed variables it was statistically demonstrated that overall CLP is more frequent in males as well as bilateral CLP whereas overall CPO is more frequent in females as well as right microform of CL. CONCLUSION: The identification of gender related subtypes of cleft is in accordance with data reported in similar studies on different populations and confirms that OFC is an heterogeneous disease even in a homogeneous ethnic group. PMID- 14765028 TI - [New aesthetic results with auricular prosthesis: two case reports]. AB - The extrinsic and intrinsic coloration of maxillo-facial silicone elastomers has always been a challenge for the clinician in order to obtain a perfect, durable integration with the surrounding skin tissues. This preliminary report describes the method for obtaining good aestethetic results for maxillo-facial prosthesis. This study shows the aesthetic results of a new silicone for the laboratory technique of creating color of the basis for a maxillo-facial prosthesis, with a good matching with human skin. The technique is useful for the direct extrinsic coloration of the surface onto the skin of the patient by means of the RTV version of the same silicone. The technological properties of this material also make it easy to reproduce the emerging profile of the prosthesis from the human skin, avoiding anaesthetic overcounturings. The 12 month follow-up shows excellent and stable aesthetic results, which are not influenced by home care procedures and by the weathering effect on color and on physical properties. PMID- 14765029 TI - Fractures of the iliac crest following anterior and posterior bone graft harvesting. Review of the literature and case presentation. AB - Iliac crest is the most commonly used extra-oral donor site of autologous non vascularized bone, because of the great amount of bone available and easy access. Bone grafting from the ileum, however, is not without complications. An extensive review of the literature focusing on fractures of the iliac crest and pelvic ring instability due to anterior and posterior bone graft harvesting is reported, and 1 case of anterior stress fracture of the iliac crest is described. Thirty-five of the 37 fractures described up to date in literature were divided in 2 groups depending on the region of bone graft harvesting. Twenty-four fractures - including our case - were related to bone graft harvesting from the anterior region, 12 were due to harvesting from the posterior region. Four out of 24 anterior fractures required further surgical treatment (16.6%). In 8 of the 12 fractures with pelvic ring instability due to posterior bone harvesting, 1 or more additional surgical procedures were performed in order to stabilize multiple fracture sites (66.6%). Anterior iliac crest fractures, even though painful, remain stable and heal spontaneously in most cases without further complications. On the contrary, fractures due to posterior iliac crest harvestings very often require complex surgical treatments and lead to significant disability, which can be permanent. PMID- 14765030 TI - Odontogenic tumor with prominent clear cell component misdiagnosed as pleomorphic adenoma by fine-needle aspiration. A case report. AB - Clear cell tumors in the maxillofacial region, are usually originated in salivary or odontogenic tissues, or may be metastatic. They include calcifying epithelial odontogenic tumors, ameloblastoma and odontogenic carcinoma. Clear cell odontogenic tumor has been classified in the last WHO classification as a benign tumor, but current opinion is that it should be designated as a carcinoma. We report a case of clear cell odontogenic tumor documented by histology, in a 82 year-old female, misinterpreted as pleomorphic adenoma by fine-needle aspiration cytology. PMID- 14765031 TI - [Skeletal and occlusal alterations in the diagnosis of Marfan syndrome]. AB - The Marfan syndrome is an autosomal dominant hereditary connective tissue disorder with variable expressivity. The incidence is estimated to be at least 1 case per 10000 individuals in most populations. The syndrome is caused by mutations in the gene coding for Fibrillin-1 (FBN1), an extracellular matrix glyco-protein. The gene responsible for the mutations was identified in the chromosome 15q21.1 region. Presented for the first time in 1896 by Dr Antoine Bernard Marfan, it was subsequently included among hereditary disorders of the connective tissue. Nowadays the Marfan syndrome is considered to be a heterogeneous pathology, that can derive from many mutations in the gene coding for Fibrillin-1. The ocular, cardiovascular and skeletal manifestations present in the individual affected by Marfan syndrome are consistent with a defect in the gene coding for a structural component of the connective tissues. From the different expressivity derive 4 types or variants of the syndrome: a) asthenia; b) non-asthenia; c) arachnodactyly; e) abnormal joint mobility. In this study the authors show that radiography and cephalometry analysis are important as diagnosis techniques in order to make an early diagnosis of Marfan syndrome, carry out differential diagnosis with other maxillo-facial pathologies and correct the therapeutic approach (orthodontic or surgical) of this syndrome. PMID- 14765032 TI - [Arrhythmic risk in essential hypertension: late potentials]. AB - AIM: A higher incidence of cardiac death exists in patients with essential hypertension, and it is higher still in those with ventricular arrhythmia. The purpose of noninvasive diagnostic imaging in hypertensive patients is to determine those with a greater risk for arrhythmia. In previous studies on hypertension, one of the inclusion criteria is diastolic blood pressure <95 mmHg, which, however, is a low selectivity criterion. Instead, our study emphasizes the need to evaluate the incidence of ventricular arrhythmia in hypertensive patients not yet receiving drug therapy and to formulate the diagnosis based on 24-h ambulatory arterial blood pressure monitoring, which represents a more selective criterion than the diastolic pressure value proposed by the World Health Organization (WHO). METHODS: A total of 128 consecutive patients with essential hypertension classified according to WHO criteria underwent 24-h monitoring, 85 (66.4%) of which presented with a mean 24-h arterial pressure >135/85 mmHg. These patients were then evaluated using mono- and two-dimensional echocardiography and 24-h dynamic Holter monitoring to detect arrhythmias and the presence of left ventricular later potentials. RESULTS: Left ventricular hypertrophy was present in 60 (70.6%) patients and absent in 25 (29.4%). Based on the Lown classification of ventricular arrhythmia, 20 (23.5%) patients had Grade I arrhythmia, 5 (5.9%) Grade II, 4 (4.7%) Grade III, 9 (10.6%) Grade IVA, 20 (23.5%) Grade IVB, 12 (14.1%) Grade V, 15 (17.6%) clinically unremarkable arrhythmia, and 17 (20%) had late potentials because they tested positive to at least 2 out of three criteria, and 2 patients were positive to all 3 criteria. CONCLUSION: Our study findings demonstrated a significant correlation between left ventricular hypertrophy and grade of arrhythmia (r=0.552; p<0.0001) and late potentials (r=0.405; p<0.001). The presence of late potentials was also found to correlate significantly with grade of arrhythmia (r=0.593; p<0.001). These patients present with a more severe stage of the disease and should therefore receive more aggressive treatment to prevent sudden cardiac death resulting from arrhythmia. PMID- 14765033 TI - Multidetector row computed tomographic angiography of the abdominal aorta and lower limbs arteries. A new diagnostic tool in patients with peripheral arterial occlusive disease. AB - AIM: The aim of this study is to evaluate the accuracy of multidetector row CT angiography (MDCTA) of the abdominal aorta and lower extremities arteries in patients with peripheral arterial occlusive disease who did not receive any prior treatment whether interventional or surgical. METHODS: Twenty-two patients with peripheral vascular occlusive disease (16 male, 6 female, age range 44-85 years) underwent MDCTA of the abdominal aorta and lower extremities. Digital subtraction angiography (DSA) of the same districts was performed within 3 months. Images were blindly interpreted by 2 interventional radiologists and compared with the results of digital subtraction angiography. RESULTS: Sensitivity and specificity of MDCTA were 92% and 94%, respectively, with positive and negative predictive values of 93% and 95%. Overall diagnostic accuracy was 93%. Normal arterial segments and 100% occlusions were correctly identified in all cases by MDCTA. Moderately stenotic segments interpretation in the calves appeared to be more controversial, but no statistical difference in accuracy in this district was noted with respect to accuracy in more proximal arteries. CONCLUSION: MDCTA of the abdominal aorta and lower extremities is a feasible, accurate imaging modality in clinical practice when compared to DSA. PMID- 14765034 TI - [Stress imaging in coronary artery disease: state of the art]. AB - To date, several diagnostic tools allow an accurate non-invasive evaluation of coronary artery disease; this is due to the great progress in echocardiographic and nuclear imaging techniques in the last 10 years. The large availability of different stress imaging techniques allows to choose the most appropriate technique for each patient according to the clinical characteristics. This paper presents the state of the art of echocardiographic and nuclear stress imaging for the diagnosis of coronary artery disease and for the prognostic stratification of infarcted patients. Advantages and limits of the different techniques are described rather than putting in competition echo and nuclear cardiology as has often been done in the past. Cardiologists should select among the various techniques on the basis of clinical characteristics of single patients, center's experience and an objective evaluation of economical aspects. PMID- 14765035 TI - Cardiopulmonary exercise testing and exhaled nitric oxide in the assessment of patients with mitral stenosis. AB - AIM: In patients with mitral stenosis, symptoms do not always correlate with echocardiographic data. The aims of the study were to evaluate the role of cardiopulmonary exercise testing in the assessment of patients with mitral stenosis and to quantify nitric oxide production at rest and at the end of exercise. METHODS: We evaluated 43 patients with moderate to severe mitral stenosis with a discrepancy between echocardiographic data and symptoms. Nitric oxide output was calculated by measuring nitric oxide concentration in the exhaled air at rest and at the end of exercise test. RESULTS: Patients were divided in 2 groups: group 1 with a functional capacity <75% at cardiopulmonary exercise test (VO2max in % of the predicted one) and group 2 with functional capacity >75%. Transvalvular gradient and pulmonary artery pressure were significantly higher in group 1 than in group 2 (respectively 9.07 +/- 2.11 mmHg vs 6.01 +/- 1.08 mmHg, p<0.001 and 42.8 +/- 7.2 mmHg vs 33.1 +/- 4.7 mmHg, p<0.001). Patients of group 1 had a lower nitric oxide output at the end of exercise compared to group 2 (231.4 +/- 96.6 nl/min vs 326.3 +/- 74.0 ml/min, p=0.01) and to normal subjects (511.15 +/- 180.1 nl/min, p<0.001). CONCLUSION: Cardiopulmonary exercise testing provides objective non invasive information in the evaluation of patients with discrepancy between symptoms and echocardiographic data. Different levels of nitric oxide output during exercise suggest the role of nitric oxide in regulating pulmonary vascular tone. PMID- 14765036 TI - Ibutilide for cardioversion of atrial flutter: efficacy of a single dose in recent-onset arrhythmias. AB - AIM: Ibutilide is particularly effective in rapid termination of atrial flutter (AFL) with few adverse effects. Despite the recommendation of 2 infusions, cardioversion may occur up to 70 minutes after a single dose. We investigated the feasibility, efficacy and safety of a single dose ibutilide treatment of AFL in a single-center, observational study. METHODS: Fifty-nine consecutive patients (44 males, mean age 70 +/- 12), referred to our CCU for paroxysmal AFL (mean arrhythmia duration 10 +/- 18 days), were treated with 1 mg ibutilide. In case of inefficacy, a 2nd dose could be administered 10 to 60 minutes later on the basis of the ECG examination for QTc prolongation and AFL cycle variations. Successful cardioversion was defined as sinus rhythm (SR) restoration within 2 hours. RESULTS: Forty-four patients (75%) converted to SR after ibutilide, 31 with single dose (53%, Group 1), and 13 with double dose (22%, Group 2). AFL duration was shorter in Group 1 (4 +/- 5 vs 16 +/- 29 days). The mean time to the 2nd dose administration was 34 +/- 11 minutes in responders, 51 +/- 23 minutes in non responders. Only 3 (5%) significant adverse events, all observed after a single dose, occurred. CONCLUSION: Ibutilide is highly effective and safe, in a monitored environment, for rapid termination of AFL. Recent onset AFL may be terminated with a single infusion in many cases; yet, cardioversion should be waited for no more than 30 minutes after the end of the 1st dose, before administering the 2nd one, in order not to reduce the possibility of SR restoration. PMID- 14765037 TI - [Effectiveness of Troxerutin in association with Pycnogenol in the pharmacological treatment of venous insufficiency]. AB - AIM: The purpose of our study was to assess the different effectiveness of Troxerutin in association with Pycnogenol compared to the effects obtainable with the same pharmacological principle in monotherapy in patients suffering from venous insufficiency. METHODS: Seventy patients with venous insufficiency of the lower extremities at the first stages of the CEAP classification (Cs1,3; Es; As 1; invalidity score 2, clinical score 1-2) were selected and subdivided into 2 groups: one of 50 and one of 20. All patients reported the following symptoms either associated or in isolation: heaviness, cramps, pruritus and pain at palpation. Evaluation of the degree of venous insufficiency was carried out by means of echo-Doppler at recruitment. The 1st group received Troxerutin (470 mg) associated with Pycnogenol (20 mg) in a dose of 1 g/day per os for 60 days; the 2nd group Troxerutin alone (300 mg) in a dose of 2 tablets twice a day per os for 60 days. Controls were set up at 30 and 60 days from the start of treatment and after a period of 3 months from the end of treatment to assess the effects on the symptomatology over time. The effectiveness of the drugs on symptomatology reported by both groups was assessed by means of an analysis of the qualitative variation of the symptoms using the score-scales method: 3=severe; 2=moderate; 1=slight; 0=absent and this variation was expressed as a percentage decrease and analysed using Student's test. RESULTS: The results of our study after 30 days of treatment highlighted a clinical improvement in all patients with the disappearance of symptoms (score from 3 to 1) in 50% of patients in the 1st group and in 35% in the 2nd with a peak of 96% in patients of the 1st group and 80% in the 2nd at the end of treatment (60 days). This result held stable in the course of follow-up for 96% of patients in the 1st group (p<0.001) and for 50% of those in the 2nd group (p<0.005). CONCLUSION: Comparison between the 2 groups showed that the group which received Troxerutin associated with Pycnogenol reported greater therapeutic effectiveness than the control group as regards both the rapidity of disappearance of the symptoms and as regards maintenance of the cure obtained. PMID- 14765038 TI - Thallium-201 myocardial perfusion imaging in patients with systemic lupus erythematosus. AB - The cardiovascular system is frequently affected in systemic lupus erythematosus (SLE). The observation of clinical manifestations related to the presence of coronary artery disease has not been frequently documented in young SLE patients. In these patients, the presence of inflammatory or thrombotic vascular lesions is often documented by anatomo-histological studies in the absence of previous clinical manifestations. The purpose of this study was to evaluate the presence of myocardial perfusion defects in SLE patients. The study was carried out in 15 patients without clinical signs of myocardial ischemia, 1 male and 14 females, 24 to 64 years old, with a mean SLE duration of 10.2 +/- 7.5 years. All the patients had normal blood pressure; electrocardiogram and Doppler-echocardiographic analysis showed values in the normal range. All the patients underwent thallium 201 exercise stress imaging repeated 3 hours later at rest, with tomographic SPECT analysis. Exercise test was carried out until submaximal load, without induction of ST segment alterations or symptoms. Scintigraphic scan showed normal thallium-201 SPECT imaging in 11/15 patients, while the other 4 patients had a slight perfusion defect, 3 of them in the inferior segment, in 2 non reversible and in 1 reversible; 1 patient had a non reversible defect in the septal segment. These slight perfusion defects, prevalently non reversible, may sometimes be a false positive imaging. Our results are in contrast with the literature observations concerning the frequent incidence of thallium-201 perfusion defects in SLE patients. In young asymptomatic SLE patients, our study does not report very important data indicating myocardial ischemia and suggesting the presence of significant coronary obstruction or vasculitis. PMID- 14765039 TI - A case of carotid bifurcation anomaly discovered during a thyroidectomy operation. AB - The anatomic anomalies of the supra-aortic trunks and their branches are comparatively frequent observations. They often remain asymptomatic throughout life although in some cases, especially if nothing is known of them, they may complicate surgical interventions involving that anatomical region. The case of a female patient who, during thyroidectomy, suffered the ligature of her external carotid artery which was taken mistakenly for the thyroid, is reported. Dissection of the area made it possible to identify an anomaly of the vascular structures with a carotid bifurcation which turned out to be very low, a very deep internal carotid artery on the prevertebral fascia and an absent lower thyroid artery. In the postoperative period, the patient was subjected to MR of the neck. This showed an anatomical anomaly of the left carotid bifurcation which was much lower than normal. An attempt was made to reconstruct the external carotid artery with a saphenous vein but unsuccessfully. The incident did not have serious complications for the patient but it might be asked whether, given the low cost and the absolute non-invasiveness of the examination, it might not be worth while to carry out a duplex scan of the supra-aortic trunks before going ahead with any operation in the cervical region, in consideration above all of the dangerous nature of these vascular structures and of the usefulness of studying pathologies that often remain silent until the occurrence of an event which might well be catastrophic. PMID- 14765040 TI - Asymptomatic intramural aortic hematomas in different clinical settings. PMID- 14765041 TI - Pain in day surgery. PMID- 14765042 TI - Pain control in day surgery: SIAARTI guidelines. PMID- 14765043 TI - Cardiac assist devices. Technology and applications. PMID- 14765044 TI - Analgesic transition after remifentanil-based anesthesia in neurosurgery. A comparison of sufentanil and tramadol. AB - AIM: Transition from the end of remifentanil infusion and postoperative analgesia must be planned carefully owing to remifentanil's (R) rapid offset. Intraoperative morphine has been used for the transition to postoperative analgesia following remifentanil-based anesthesia. Sufentanil (S) is a very potent opioid with high micro-receptor affinity, a much wider therapeutic index and a lower fractional receptor occupancy. These pharmacological and dynamics features make sufentanil an interesting alternative to morphine for immediate postoperative analgesia. METHODS: EXPERIMENTAL DESIGN: perspective, randomized, single blinded and comparative study. Institution: neurosurgical operating theatre at University. PATIENTS: 96 patients, aging from 25 to 67 years, ASA class I-III, undergoing neurosurgical operations, were studied. INTERVENTIONS AND MEASUREMENTS: the anesthetic management was: premedication: atropine 0.01 microg kg(-1) + remifentanil 0.20 microg kg(-1) min(-1); induction: propofol 2.0 microg kg(-1) + cisatracurium 0.15 microg kg(-1); maintenance: sevoflurane 0.8% + remifentanil (titrated infusion) cisatracurium. All patients received ketorolac 30 mg i.v. 1 hour before the end of surgery and ketorolac (60-90 mg) + tramadol (200-300 mg) by elastomeric pump; patients were divided into 2 groups: group T receiving tramadol 100 mg and group S receiving a bolus dose of sufentanil 0.10 microg kg(-1), 30 and 15 minutes before the end of surgery respectively. Recovery time, postoperative analgesia evaluated by VAS, cardiocirculatory parameters and side effects like nausea, vomiting, shivering, muscle rigidity, sedation and respiratory depression were recorded. RESULTS: VAS was significantly lower in Group S. Recovery time was shorter in Group T than in Group S (8.8 +/- 3.6 vs 11.6 +/- 4.6 min), no statistically significant differences between groups as regards nausea, vomiting and shivering. Short-lasting respiratory depression was detected in 3 cases in Group S. CONCLUSION: At the emergence much better control of the transition phase in patients treated with sufentanil: smooth recovery with better tolerability of the endotracheal tube; efficacious analgesia along with cardiocirculatory stability. PMID- 14765045 TI - Deep sedation for magnetic resonance imaging. Personal experience. AB - AIM: Precision in diagnostic procedure and examination of paediatric patients often requires their absolute immobility. Deep sedation has proven to be an excellent method, allowing optimum technical quality of MRI particularly in younger age groups. The aim of study is to demonstrate the possible application of deep sedation through the use of 2 safe and manageable drugs. METHODS: We carefully evaluated and selected 82 patients (47 males and 35 females; average age 5.4 years): they came from various paediatrics departments. Deep sedation was practiced with: Chloral hydrate (60-80 mg/kg in one oral administration); propofol as intravenous bolus (2-2.5 mg/kg) followed by a maintenance infusion of 75-125 microg/kg/min. This was preceded by midazolam (0.05 mg/kg i.v.) outside the MRI room. Oxygen saturation (SpO2) was monitored in all patients along with heart rate in order to foresee the need for any possible therapeutic intervention. RESULTS: The sedation levels attained permitted the success of MRI assuring the immobilization required. Manually assisted mask ventilation was required for a period of 2-3 min in 5 patients treated with propofol. All other patients breathed autonomously. Complete reawakening occurred within 2 hours of drug administration. Surveillance was prolonged inside their respective units, however, without registering delayed side effects. CONCLUSION: The central point of the success of deep sedation is to define the type and dose of optimum drug for individual patients. This requires a qualified, expert group ready to intervene in the presence of adverse results of drugs administered. Propofol and chloral hydrate are the optimum drugs for diagnostic techniques requiring total immobilization and rapid reawakening. PMID- 14765046 TI - Candidemia in Intensive Care patients. Risk factors and mortality. AB - AIM: Aim of this study was to evaluate if the risk factors for candidemia could be used to identify patients who have a greater possibility of death after Candida spp blood infection. METHODS: A retrospective observational comparative study. SETTING: the Intensive Care Unit of an University Hospital. PATIENTS: 478 critical patients were included in this study. Neutropenic and immuno-suppressed patients were excluded. INTERVENTIONS: routine care for acutely ill patients, with regard to their pathology. MEASUREMENTS: age, APACHE II at the admission, length of stay in the ICU before the diagnosis of candidemia and whole length of stay, outcome, risk factors for candidemia (Candida colonisation, previous antibiotic therapy, central vein, mechanical ventilation, abdominal surgery, hemodialysis, adult respiratory distress syndrome, chronic obstructive pulmonary disease, diabetes, malignancy, splenectomy, immunosuppression, total parenteral nutrition, malnutrition) and clinical signs of multiorgan failure, systemic inflammatory response syndrome, sepsis or shock, concomitant presence of other infections. RESULTS: Twelve Candida spp blood infections were diagnosed. All the risk factors were homogenously distributed between patients who survived and those who died with the exception of the malnutrition state, associated with a higher mortality rate. CONCLUSION: If the candidemia is present, none of the risk factors for the onset of fungemia considered in this study, but the malnutrition state, are mortality predictors. PMID- 14765047 TI - Therapeutic Intervention Scoring System-28 as a tool of post ICU outcome prognosis and prevention. AB - AIM: To examine the effects of severity upon discharge from Intensive Care Unit (ICU) status, as assessed by the Therapeutic Intervention Scoring System-28 (TISS 28) on subsequent post ICU outcome. METHODS: One-year retrospective observational study. Six bed general ICU in a general hospital with no High Dependency ICU unit (HDU) available. We used data from all patients admitted to the ICU. From all discharged patients, data on mean length of ICU stay, APACHE II upon admission and TISS-28 upon discharge were collected. RESULTS: Eighty-six patients, mean age 63.2, were discharged to hospital wards between January 1999 and December 1999. Age is a significant factor to contribute to outcome prediction, (p=0.0478). TISS 28 is statistically significant related to survival status. Thirteen patients that did not survive had higher TISS-28 values (p=0.0032). Length of ICU stay has a borderline association (p=0.063) with survival. CONCLUSION: Patients discharged from ICU have post ICU hospital length of stay and prognosis related to their age, time of hospitalization in ICU and their severity status upon discharge from the ICU. We believe that, based on the TISS-28 scoring system, high risk patients can be identified and either ideally transferred to a HDU or discharged from ICU when further improvement has been achieved. TIS-28 is a valuable tool in post ICU outcome prediction and subsequently, in hospital mortality reduction. At the same time, through TISS-28 use, each hospital can locally identify the quality of care provided outside the ICU, given the outcomes measured in groups of patients at various severity levels. PMID- 14765048 TI - Cytopathologic examination of epidural catheter for postoperative analgesia. Pathophysiology and clinical management. AB - AIM: The authors performed a prospective study in a series of patients undergoing combined general and epidural anaesthesia for major abdominal surgery in order to define if the epidural catheter inserted for postoperative analgesia induced in the short-term (7-8 postoperative days) any cytopathologically appreciable inflammatory response. METHODS: From April to September 2001, 20 consecutive patients undergoing combined general and epidural anaesthesia for major abdominal surgery at the National Cancer Research Institute and Villa Scassi Hospital (Genoa), were recruited after obtaining Institutional Ethics Committee approval and written consent from the patients. The standard technique for epidural anaesthesia was adopted. Preoperatively, all patients received peridurally a dose test of 3 ml of 2% lidocaine (60 mg) followed by 5 ml of ropivacaine 0.75%, and a continuous infusion of ropivacaine 0.375% (5-10 ml/h; maximal dose=20 ml) intraoperatively. As regards the therapeutic management of postoperative analgesia, patients received a continuous infusion of ropivacaine 0.2% for at least 48 hours and supplemental bolus (2 mg/die) of morphine hydrochloride. The epidural catheter was always removed between the 7th and 8th postoperative day, and it was examined by the pathologist according to the Thin Prep 2000 procedure. RESULTS: The cytopathologic examination of the tip of the epidural catheter gave the following findings: amorphous material without cells (n=10); rare granulocytes and histiocytes (n=6); stromal cells (n=3), and rare lymphocytes (n=1). CONCLUSION: We were unable to detect any cytopathologically appreciable inflammatory response at the tip of the epidural catheter which could have suggested the occurrence of inflammation in the epidural tissues. Given the positive results of prophylactic epidural administration of small doses of corticosteroids in the reduction of postepidural anaesthesia back pain and their direct membrane action on nociceptive C-fibers, this kind of backache seems to be related to the stimulations of such nociceptors more than to a catheter-related inflammatory response of epidural tissues with possible evolution in peridural fibrosis, as reported following surgical intervention for lumbosacral disease. PMID- 14765049 TI - Treatment of missing lateral incisors. PMID- 14765050 TI - Distal movement of mandibular molars in adult patients with the skeletal anchorage system. AB - The skeletal anchorage system (SAS) consists of titanium anchor plates and monocortical screws that are temporarily placed in either the maxilla or the mandible, or in both, as absolute orthodontic anchorage units. Distalization of the molars has been one of the most difficult biomechanical problems in traditional orthodontics, particularly in adults and in the mandible. However, it has now become possible to move molars distally with the SAS to correct anterior crossbites, maxillary dental protrusion, crowding, and dental asymmetries without having to extract premolars. This study evaluated the treatment and posttreatment changes during and after distalization of the mandibular molars. In 15 adult patients (12 women and 3 men), a total of 29 mandibular molars were successfully distalized with SAS. The amount of distalization and relapse and the type of tooth movement were analyzed with cephalometric radiographs and dental casts. The average amount of distalization of the mandibular first molars was 3.5 mm at the crown level and 1.8 mm at the root level. The average amount of relapse was 0.3 mm at both the crown and root apex levels. Of 29 mandibular molars, 9 were tipped back, and the others were translated distally in accordance with the established treatment goals. SAS is a viable modality to move mandibular molars for distally correcting anterior crossbites, malocclusions characterized by mandibular anterior crowding, and dental asymmetries. PMID- 14765051 TI - Long-term evaluation of sandblasted and acid-etched implants used as orthodontic anchors in dogs. AB - The aim of this study was the clinical, radiographic, and histologic evaluation of the tissue surrounding SLA (sandblasted and acid-etched) implants loaded with a continuous and constant force for 52 weeks, after a healing period of 6 weeks, after implant insertion. SLA implants were placed in the maxilla of 3 dogs and the mandible of 5 dogs after a 12-week healing period after extraction. Abutments were attached to the 6 test implants (2 in the maxilla, 4 in the mandible). Superelastic nickel-titanium coil springs were activated between the SLA implants and the canines, producing a force of 200 g (2 N). Two unloaded implants (1 in the maxilla, 1 in the mandible) served as controls. Histologic analysis showed a corticalization of bone trabeculae, thicker at the loaded than at the unloaded implants. New bone formation at the level of the crest was slightly superior in the test implants. A difference between the tension and compression areas could not be observed in the test implants. The mean bone-implant contact values of the test implants for the maxilla and mandible were 40.23% and 49.33%, respectively. In the control implants, the bone-implant contact value was 67.91% for the maxilla and 49.23% for the mandible. SLA implants can be used as an anchorage unit with confidence, in spite of a short healing period and a prolonged force application. Further studies with different force magnitudes and healing periods are required to clarify the effects of healing period and force magnitude on bone quality and quantity. PMID- 14765052 TI - Comparison of maxillary and mandibular growth. AB - In this longitudinal study, serial lateral cephalometric radiographs were used to compare growth patterns of the maxilla and mandible, with hand-wrist radiographs used to assess skeletal maturity. The sample comprised 28 untreated subjects (15 female, 13 male) who were followed from ages 6 to 20 years. All subjects had Class I malocclusions without anterior crossbites. Absolute values and incremental changes for linear and angular cephalometric measurements were recorded and analyzed, and the relative growth-rate formula was used to provide an accurate index of acceleration and deceleration of growth. The SNA angle did not change significantly with age, but the SNB angle increased significantly in the male subjects. The ANB angle decreased continuously until age 14. The palatal plane descended significantly from the horizontal plane. The anterior and posterior nasal spines moved at about the same rate. The mandible grew in length twice as much as the maxilla from ages 6 to 20. With growth, the facial profiles of the male subjects became straighter as the chin became more prominent. The female subjects had less incremental growth and duration of growth of the mandible, so that the profiles remained more convex. Overall, skeletal and chronologic ages did not differ significantly, except at ages 10 and 16 in the female subjects. Individual variability pointed to the need for assessing each patient's pattern in the general guidelines of the group pattern. PMID- 14765053 TI - Relationship between maxillofacial morphology and deglutitive tongue movement in patients with anterior open bite. AB - We reported previously that patients with anterior open bite had tongue tip protrusion, slower movement of the rear part of the dorsal tongue, and earlier closure of the nasopharynx during deglutition. In the present study, the relationship between this characteristic tongue movement and maxillofacial morphology in patients with anterior open bite was investigated. The subjects were 10 female patients with anterior open bites and 10 women with normal overbites as controls. Maxillofacial morphology was measured by cephalometric radiography, and tongue movement during deglutition was analyzed by cineradiography. The relationship between each value obtained by cephalometric radiography and cineradiography was evaluated by simple correlation analysis. In the patients with anterior open bite, there were significant correlations between mandibular plane angle, ramus height of the mandible, or anteroposterior dimension of the maxilla and movement of the front part of the dorsal tongue during deglutition. Furthermore, there were also significant correlations in these patients between mandibular plane angle, gonial angle, or ramus height of the mandible and the change in the contact between tongue and palate during deglutition. The controls did not have the correlations like these. Our study suggests that characteristic tongue movements during deglutition in patients with anterior open bites are closely related to their morphological features. PMID- 14765054 TI - Effects of continuous and interrupted orthodontic force on interleukin-1beta and prostaglandin E2 production in gingival crevicular fluid. AB - The purpose of this study was to evaluate the effects of a light continuous force and an interrupted force with weekly reactivation on interleukin-1beta (IL-1beta) and prostaglandin E(2) (PGE(2)); possible interactions between these 2 potent mediators of the bone resorption process were assessed in vivo. Ten healthy young adults (mean age 20.6 years, 2 men, 8 women) with 4 premolars extracted were assessed. In each subject, 1 maxillary canine (E1) received continuous force with a nickel-titanium coil spring. The opposite canine (E2) received an interrupted force with a screw-attached retractor; the force was reactivated weekly by 2 turns of the screw. An antagonistic canine was used as a control. Gingival crevicular fluid was collected from the distal side of each tooth, 10 times in 3 weeks, and IL-1beta and PGE(2) levels were measured. For E1, the IL-1beta level showed a significant elevation at 24 hours and then decreased and maintained an insignificant but high mean concentration, compared with the control site. The PGE(2) level showed a significant elevation at 24 hours and then decreased. For E2, a significant elevation of IL-1beta level was observed at 24 hours and a greater significant elevation at 24 hours after the first reactivation, compared with the control sites. The PGE(2) level increased significantly at 24 hours and remained high for 1 week. The synergistic up-regulation of PGE(2) by appliance reactivation and secreted IL-1beta was not evident with either type of force after 1 week. Both experimental sites showed significant tooth movement compared with the control sites at 3 weeks; however, there was no significant difference between the 2 experimental sites. A well-controlled mechanical stress with timely reactivation can effectively upregulate IL-1beta secretion, but there might be limitations in increasing the mediator levels, because of the feedback mechanisms in vivo. In addition, the analysis of crevicular fluid is a useful method for assessing cellular response to orthodontic force in vivo. PMID- 14765055 TI - Immunohistochemical localization of alphavbeta3 integrin receptor during experimental tooth movement. AB - During orthodontic treatment, multinucleated clast cells carry out the resorption of mineralized tissues. Adhesion of clast cells to the mineralized tissues is mediated by transmembrane cell-surface glycoproteins called integrins, specifically by the alphavbeta3 integrin, which plays an important role in the process of bone resorption. The role of the alphavbeta3 integrin in bone resorption leading to osteoporosis has been demonstrated, but its role in alveolar bone and root resorption during orthodontic tooth movement is unknown. This study examined the expression of the alphavbeta3 integrin during experimental tooth movement. Tooth movement was achieved in 16 male Sprague Dawley rats (each weighing 120-200 g) with elastic bands between their maxillary first and second molars. The molar-bearing segments were dissected and processed for histologic and immunohistochemical examination. The expression of alphavbeta3 integrin was examined with 2 primary antibodies: a polyclonal anti-alphav integrin subunit antibody and a polyclonal anti-beta3 integrin subunit antibody. Negative controls were similarly processed but without incubation with primary antibodies. The alphavbeta3 integrin was expressed both by osteoclasts associated with alveolar bone resorption and by odontoclasts associated with root resorption during experimental tooth movement. Furthermore, the beta3 integrin subunit was expressed by the epithelial rests of Malassez in the periodontal ligament. Negative controls did not show immunolabeling. The alphavbeta3 integrin adhesion receptor is expressed during experimental tooth movement and might be involved in the process of mineralized tissue resorption and the functions of the epithelial rests of Malassez. PMID- 14765056 TI - Vascular endothelial growth factor expression and bone formation in posterior glenoid fossa during stepwise mandibular advancement. AB - This study assessed the amount of vascular endothelial growth factor (VEGF) expression and related the findings to new bone formation in the posterior glenoid fossa during stepwise mandibular advancement. A total of 250 female Sprague-Dawley rats, 35 days old, were randomly divided into 10 groups, each including 5 control and 20 experimental rats. Within each group, 10 experimental rats were fitted with functional appliances with a 1-step advancement of 3.5 mm. Another 10 were fitted with stepwise appliances with an initial advancement of 2 mm and a subsequent increase to 3.5 mm on day 30. The rats in the experimental groups were killed on days 3, 7, 14, 21, 30, 33, 37, 44, 51, and 60, respectively. The matched controls were killed on the same time points. Sections (7 microm) were cut through the glenoid fossa sagittally and stained with anti VEGF antibody. VEGF expression in the posterior glenoid fossa was evaluated with a computer-assisted image-analyzing system. Both VEGF expression and new bone formation were greater in the experimental rats than in the controls. During stepwise advancement, initial VEGF expression was less than that of 1-step advancement, but the second advancement elicited another peak on day 44. New bone formation was also less than that of 1-step advancement during early stages of stepwise advancement but then began to increase from day 37 onward. The maximum increase was observed on day 60. Stepwise advancement of the mandible delivers mechanical stimuli that produce a series of tissue responses that lead to increased vascularization and bone formation. PMID- 14765057 TI - Effect of titrated mandibular advancement and jaw opening on the upper airway in nonapneic men: a magnetic resonance imaging and cephalometric study. AB - The aim of this study was to examine adaptive changes in the cross-sectional area of the upper airway during mandibular advancement and jaw opening and to explore whether these changes are directly related to morphologic features of the mandible. Fourteen nonapneic men participated in this study. A custom-made oral device was used to keep the mandible at 0% (F(0)V(0)), 50% (F(50)V(0)), 75% (F(75)V(0)), or 100% (F(100)V(0)) of maximum advancement, and at 50% (F(75)V(50)), 75% (F(75)V(75)), or 100% (F(75)V(100)) of maximum gape at F(75)V(0). Magnetic resonance imaging was used to investigate differences in the upper-airway cross-sectional area in these 7 positions. To eliminate excessive individual variation, changes in the mean (mean%) and minimum (min%) cross sectional area of the upper airway were calculated as percentages of those at F(0)V(0). Cephalograms were taken to analyze morphologic features of the mandible. A dose-dependent change was seen when the mandible was advanced. The min% in F(100)V(0) was significantly greater than those in F(50)V(0) and F(75)V(0). The change in the upper airway size varied between subjects during jaw opening. At F(75)V(0), the min% of the whole upper airway was significantly associated with morphologic features of the mandible, such as the mandibular plane angle and lengths of the mandibular body and ramus. PMID- 14765058 TI - Eruption of third molars: relationship to inclination of adjacent molars. AB - The purpose of this study was to determine the relationship between the sagittal inclination of the first and second maxillary molars and the eruption of the third molars. The sample consisted of 2 groups. The subjects in the first group (n = 28) had complete normal dentitions including third molars; those in the second group (n = 32) had impacted right and left third molars. The sagittal inclinations to the palatal plane of the first and second maxillary molars were measured on computed tomography sagittal images obtained with multiplanar reconstructions. The Mann-Whitney U test was used to compare mean angular values between the 2 groups. Spearman correlation coefficients were calculated to assess the relationship with age. A multivariate analysis was used to evaluate the relationship between the eruption of the third molars and the sagittal inclination of the first molar, the second molar, sex, and age. Maxillary first and second molars were more mesially inclined in the first group, particularly in the younger subjects (16-25 years). This inclination increased with age. The logistic regression showed that the sagittal inclination of the first molar is a predictor of the eruption of the third molar. This finding suggests that a more mesially inclined maxillary dentition is likely to be associated with third molar eruption. The absence of data on space requirements in the maxillary arch and interarch relationships warrants further exploration in an orthodontic population. PMID- 14765059 TI - Time-saving fixed lingual retainer using DuraLay resin transfer. PMID- 14765060 TI - Interdisciplinary treatment including forced extrusion and reintrusion of a traumatized mandibular incisor for a patient with Class II Division 1 skeletal open bite. AB - A woman with Class II Division 1 long-face syndrome characteristics had a history of facial trauma, dentoalveolar fracture of her mandibular anterior teeth, and temporomandibular joint pain. The pretreatment apical radiograph showed a large area of external root resorption of the mesial surface of the mandibular left central incisor. To arrest the external root resorption, the mandibular left central incisor was extruded. During extrusion, sequential apical radiographs were taken. As the tooth moved away from the site of osteoclastic activity, resorption ceased, and repair took place on the root surface. At this time, the mandibular left central incisor was intruded, the mesial defect self-repaired, the tooth remained vital, and the periodontial ligament was intact. The biologic bases for this cessation of resorption and the repair of the tooth's surface are presented. PMID- 14765061 TI - Image editing. PMID- 14765062 TI - What does the histogram of an image show? PMID- 14765063 TI - Litigation, legislation, and ethics. Determining a national standard of care. PMID- 14765064 TI - The constant challenge of change. PMID- 14765065 TI - Universal respiratory etiquette: a modest proposal. PMID- 14765066 TI - Tribute to military nurses & correct Army nurses' E-mail address. PMID- 14765067 TI - Tribute to military nurses & correct army nurses' E-mail addresses. PMID- 14765068 TI - Open letter from ENA to ER producers. PMID- 14765069 TI - One mother's solution to the nursing shortage. PMID- 14765070 TI - Emergency nurse veteran appreciates ENA support. PMID- 14765071 TI - Avoid lawsuits: Warn patients that medication may make them drowsy and not to drive. PMID- 14765072 TI - Abusive physicians: violence is not "part of the job". PMID- 14765073 TI - Abusive physicians: violence is not "part of the job". PMID- 14765074 TI - Assessment and management of patients with implanted intrathecal pumps: ED clinical policy sought. PMID- 14765075 TI - Cultural differences change pain scale ratings in Togo. PMID- 14765076 TI - A 73-year-old woman with rhabdomyolysis induced by simvastatin (Zocor). PMID- 14765077 TI - ENA's Delphi study on national research priorities for emergency nurses in the United States. AB - INTRODUCTION: This study identified and prioritized research questions with greatest value to emergency nurses and of highest importance for health care consumers. METHODS: Three hundred twenty emergency nursing leaders were invited to participate in 3 rounds of mailed surveys aimed at developing consensus. During round I, 147 nurses submitted 456 research problems. These problems were synthesized into 154 researchable questions, encompassing 17 themes. The round II questionnaire listed these questions in random order. Respondents used a 7-point Likert scale to rate each question's value for practicing nurses and importance for health care consumers. One hundred one nurses rated 106 questions >5.0. The round III questionnaire was individualized to provide each respondent's round II score and each item's median group score. Reflecting on these data, subjects again rated the 106 questions. RESULTS: Seventy-nine nurses rated 16 questions 6.0 or greater for value for practicing nurses; 3 questions were rated 6.0 or greater for importance to consumers. Optimum staff to patient ratios, effects of mandatory overtime, holding admitted patients, and ED overcrowding, as well as effective strategies for educating and ensuring competence of nurses, were highest priority research problems for practicing nurses. Pain relief, impact of and methods of decreasing holding/lengthy ED stay, and effective strategies for patient teaching were judged most important for consumers. DISCUSSION: Nurses' concerns with staff shortages and overcrowding of emergency departments and their effects on patients are paramount. Pain management and patient education were chief clinical issues requiring research. The mission of ENA is "to provide visionary leadership for emergency nursing and emergency care." This mission encompasses a number of values, including the following: "The discipline of emergency nursing includes a defined and evolving body of knowledge based on research."(1) In 1998, the ENA appointed vision councils to develop futuristic ideas to advance ENA's mission. The Research Vision Council proposed that ENA develop a center for emergency nursing research to provide leadership and focus for scientific approaches to the many issues that concern emergency nurses and their patients. The Board of Directors supported this idea by formulating a work group to develop the National Institute for Emergency Nursing Research. In 1999, the work group proposed that a study be conducted to determine national priorities for emergency nursing research. This article reports on the methods and results of a Delphi study conducted for that purpose. PMID- 14765078 TI - The Emergency Severity Index (version 3) 5-level triage system scores predict ED resource consumption. AB - OBJECTIVES: The Emergency Severity Index (ESI) version 3 is a valid and reliable 5-level triage instrument that is gaining in popularity. A unique component of the ESI algorithm is prediction of resource consumption. Our objective was to validate the ESI version 3 triage algorithm in a clinical setting for the following outcome measures: actual resource consumption and patient length of stay in the emergency department and hospital. METHODS: We conducted a retrospective, descriptive study of 403 ED patients who presented to a large academic medical center. The following dependent variables were abstracted from the ED record: number of ED resources used and emergency department and hospital length of stay. The relationship between ESI level and each of the dependent variables was determined. RESULTS: Mean resource use decreased monotonically as a function of ESI level 1 (5), 2 (3.89), 3 (3.3), 4 (1.2) and 5 (0.2). The ED average length of stay (minutes) per ESI level was as follows: 1 (195), 2 (255), 3 (304), 4 (193), and 5 (98). ESI triage level did not predict hospital length of stay. CONCLUSIONS: The ESI algorithm accurately predicted ED resource intensity and gives administrators the opportunity to benchmark ED length of stay according to triage acuity level. PMID- 14765079 TI - Laryngeal mask airway: review of indications and use. PMID- 14765080 TI - Rhabdomyolysis following a gunshot wound and one trauma center's protocol and guidelines. PMID- 14765081 TI - A humane ED seclusion/restraint: legal requirements, a new policy, procedure, "psychiatric advocate" role. PMID- 14765082 TI - Knowledge assessment and preparation for the certified emergency nurses examination. PMID- 14765083 TI - Pediatric patient safety: common problems in the use of resuscitative aids for simplifying pediatric emergency care. PMID- 14765085 TI - Avoiding dangerous abbreviations and dose expressions. PMID- 14765086 TI - EMS relations: If you can't say something nice... PMID- 14765087 TI - Raising college students' alcohol awareness: a successful program at the University of Virginia. PMID- 14765089 TI - Unfolding case based scenarios: a method of teaching and testing the critical thinking skills of newly licensed nurses. PMID- 14765090 TI - Practical guidance for the new graduate advanced practice nurse regulations/certification. PMID- 14765091 TI - Carbon monoxide: case scenarios and update on ED therapy. PMID- 14765092 TI - Motor vehicle crash victim with chest injury. PMID- 14765093 TI - Three-year-old with fever, cough, chest/abdominal rash, and swollen fingers. PMID- 14765094 TI - Construction of a complete URA5 deletion strain of a human pathogenic yeast Cryptococcus neoformans. AB - Cryptococcus neoformans is an opportunistic human pathogen, which infects the central nervous system causing the fatal disease, meningitis. In order to understand the genetic background of this human pathogen, the basic molecular manipulation techniques of deletion, overexpression, and so on have been developed. URA5, a gene encoding orotate phosphoribosyltransferase, has frequently been used to introduce foreign gene fragments by complementing ura5 mutant strains, which are not, however, stable; reversion to uracil prototroph is thus frequently observed on selective condition. The high possibility of reversion makes it inconvenient to use this mutation to identify appropriate transformants and thus, manipulation in molecular genetics. We report here the isolation of a stable ura5 mutant of C. neoformans, designated as TAD1, by eliminating the URA5 gene by homologous recombination using the biolistic DNA delivery system. The availability of the stable ura5 mutant offers the advantage that no spontaneous reversion occurs so that a satisfactory rate of homologous recombination can be achieved. The strain will allow efficient genomic analysis in C. neoformans. PMID- 14765095 TI - [Screening Examination and Management of Dermatophytosis by Trichophyton tonsurans in the Judo Club of a University]. AB - Thirty-one members of the Judo Club of a certain university (age: 18~23) underwent a screening examination for dermatophytosis by Trichophyton tonsurans. Test items were: age, sex, height, weight, living mode, exercise duration, number of judo contestants, presence of foreign contestants, occurrence, if any, of dermatophytosis past or present according to a subject's answers to a questionnaire, medical examinations and mycological examinations (KOH, cellophane tape culture, and hairbrush culture). Twenty-four subjects (77%) replied that they had suffered from dermatophytosis in the past, and 8 subjects (26%) had had head eruption in the past. Eleven subjects (35%) had suspicious dermatophytosis at the time of screening; 3 of them were found positive by direct microscopy, 2 of them were positive by cellophane tape culture. Eleven subjects (35%) were found positive by the hairbrush culture, but only 2 had eruption-like folliculitis. The remaining 9 subjects were free from clinical symptoms and were judged to be asymptomatic carriers. As countermeasures, we recommended cleaning and the use of shampoo containing miconazole nitrate. Subjects with suspicious tinea corporis were treated with antimycotic ointment. The 7 subjects who showed more than 5 colonies by the hairbrush culture were treated with 1-week pulse therapy of 400 mg itraconazole, and 3 of these who took a total dose of a pulse became negative through one pulse therapy. PMID- 14765096 TI - [Trichophyton violaceum infection in three black brothers]. AB - Case 1 was a 3-year-old with tinea capitis, while Case 2 was a 5-year-old with kerion celsi. These conditions developed between December 1997 and January 1998 when the patients were living in Canada. Case 3 was a 7-year-old who developed kerion celsi in March 1998 in Japan. For Cases 1 and 2, the referring doctor had detected a fungal infection a few weeks previously, and griseofulvin was administered orally to Case 1 for three weeks and Case 2 for one week prior to consultation at our department. Both Cases 1 and 2 visited our department initially on May 7, 1998 with the causative agent separated from their lesions by the referring doctor respectively. The causative agent was identified as Trycophyton violaceum by our department. KOH-prepared direct microscopy and culture were positive for Case 1, but negative for Case 2. Infected hairs of Case 1 showed a chain of large arthrospores arranged in parallel rows inside the hair. Case 3 visited our department initially on May 21, and T. violaceum was also isolated from this patient's lesion. A reddish purple isolate that was stored for five months after isolation and culture was subcultured using a Sabouraud agar containing thiamin. After two months, brown colonies without red pigment were seen in the reddish purple colonies. Slide culture performed for both colonies showed intercalary and terminal chlamydospores, thus confirming that they were the same fungus. These findings appear to represent the conversion of T. violaceum into T. glabrum. PMID- 14765097 TI - First isolates of Nocardia abscessus from humans and soil in Japan. AB - Nocardia abscessus, a recently established species, was isolated from patients during 2000. In the course of our taxonomic studies on 121 clinical Nocardia isolates in Japan 5 strains isolated from patients plus one strain isolated from soil in Japan, were found to have similar physiological characteristics to those of N. abscessus. Phylogenetic studies using 16S rRNA gene sequence analysis confirmed that these strains belong to N. abscessus. This is the first isolation report of N. abscessus from soil as well as from clinical samples in Japan. PMID- 14765098 TI - Mitochondrial DNA analysis of Sporothrix schenckii from China, Korea and Spain. AB - Sporothrix schenckii isolates from China, Korea and Spain were investigated for mtDNA types based on restriction fragment length polymorphism patterns with HaeIII. Sixty-one isolates from China, 8 from Korea and 11 from Spain were comprised of 7, 2 and 2 mtDNA types, respectively. All the isolates belonged to Group B. PMID- 14765099 TI - Optical coherence tomography--a review of the principles and contemporary uses in retinal investigation. PMID- 14765100 TI - Topical timolol prevented migraine attacks. PMID- 14765101 TI - Successful management of three consecutive cases of recurrent corneal erosion with botulinum toxin injections. PMID- 14765102 TI - Squamous cell carcinoma of the frontal and ethmoidal paranasal sinuses masquerading as acute dacryocystitis. PMID- 14765103 TI - Early division of a modified Cutler-Beard flap with a free tarsal graft. AB - AIMS: We describe a variation of the Cutler-Beard flap in the reconstruction of upper eyelid defects. METHODS: The technique of upper eyelid reconstruction with a free tarsal graft and a cutaneous lower eyelid advancement flap divided at 2 weeks is described. Four cases where this technique was used for reconstruction of eyelid defects due to periocular malignancy are also reported. RESULTS: There were three male and one female patients ranging in age from 61 to 78 years. The underlying diagnoses were squamous cell carcinoma, trichilemmal carcinoma, and two basal cell carcinomas. Follow-up of 6-28 months revealed a good outcome in all four cases with one patient developing a mild cicatricial lower eyelid ectropion that was managed conservatively. CONCLUSIONS: A modified Cutler-Beard flap with free tarsal graft and early division may provide an effective alternative for upper eyelid reconstruction in cases with sufficient lower eyelid skin laxity. PMID- 14765104 TI - Corneal adherent leukoma associated with measles. PMID- 14765106 TI - Mapping key functional sites within yeast TFIID. AB - The transcription factor TFIID, composed of the TATA box-binding protein (TBP) and 14 TBP-associated factors (TAFs), plays a key role in the regulation of gene expression by RNA polymerase II. The structure of yeast TFIID, as determined by electron microscopy and digital image analysis, is formed by three lobes, labelled A-C, connected by thin linking domains. Immunomapping revealed that TFIID contains two copies of the WD-40 repeat-containing TAF5 and that TAF5 contributes to the linkers since its C- and N-termini were found in different lobes. This property was confirmed by the finding that a recombinant complex containing TAF5 complexed with six histone fold containing TAFs was able to form a trilobed structure. Moreover, the N-terminal domain of TAF1 was mapped in lobe C, whereas the histone acetyltransferase domain resides in lobe A along with TAF7. TBP was found in the linker domain between lobes A and C in a way that the N-terminal 100 residues of TAF1 are spanned over it. The implications of these data with regard to TFIID function are discussed. PMID- 14765107 TI - ATF6 modulates SREBP2-mediated lipogenesis. AB - Activating transcription factor 6 (ATF6) and sterol regulatory element-binding proteins (SREBPs) are activated by proteolytic cleavage. The ensuing nuclear translocation of their N-termini (i.e., ATF6(N) and SREBP(N)) activates the respective target genes involved in unfolded protein response and lipogenesis. Here, we report that glucose deprivation activated ATF6 but suppressed the SREBP2 regulated transcription. Overexpression of ATF6(N) had similar inhibitory effects on SREBP2-targeted genes. The blockade of ATF6 cleavage by BiP/grp78 reversed this inhibitory effect. GST pull-down and immunoprecipitation assays revealed that ATF6(N) bound to SREBP2(N). Deletion analysis of the various functional domains of ATF6 indicated that the interaction was through its leucine-zipper domain. Chromatin immunoprecipitation assays revealed that ATF6(N) formed a complex with the SRE-bound SREBP2(N). The attenuated transcriptional activity of SREBP2 was due, in part, to the recruitment of HDAC1 to the ATF6-SREBP2 complex. As a functional consequence, the lipogenic effect of SREBP2(N) in liver cells was suppressed by ATF6(N). Our results provide a novel mechanism by which ATF6 antagonizes SREBP2 to regulate the homeostasis of lipid and glucose. PMID- 14765108 TI - Recovery from DNA damage checkpoint arrest by PP1-mediated inhibition of Chk1. AB - The G2 DNA damage checkpoint delays mitotic entry via the upregulation of Wee1 kinase and the downregulation of Cdc25 phosphatase by Chk1 kinase, and resultant inhibitory phosphorylation of Cdc2. While checkpoint activation is well understood, little is known about how the checkpoint is switched off to allow cell cycle re-entry. To identify proteins required for checkpoint release, we screened for genes in Schizosaccharomyces pombe that, when overexpressed, result in precocious mitotic entry in the presence of DNA damage. We show that overexpression of the type I protein phosphatase Dis2 sensitises S. pombe cells to DNA damage, causing aberrant mitoses. Dis2 abrogates Chk1 phosphorylation and activation in vivo, and dephosphorylates Chk1 and a phospho-S345 Chk1 peptide in vitro. dis2Delta cells have a prolonged chk1-dependent arrest and a compromised ability to downregulate Chk1 activity for checkpoint release. These effects are specific for the DNA damage checkpoint, because Dis2 has no effect on the chk1 independent response to stalled replication forks. We propose that inactivation of Chk1 by Dis2 allows mitotic entry following repair of DNA damage in the G2 phase. PMID- 14765109 TI - Fission yeast Clp1p phosphatase affects G2/M transition and mitotic exit through Cdc25p inactivation. AB - The Cdc14 family of phosphatases specifically reverses proline-directed phosphorylation events. In Saccharomyces cerevisiae, Cdc14p promotes Cdk1p inactivation at mitotic exit by reversing Cdk1p-dependent phosphorylations. Cdk1p is a proline-directed kinase whose activity is required in all eukaryotes for the transit into mitosis. At mitotic commitment, Cdk1p participates in its own regulation by activating the mitotic inducing phosphatase, Cdc25p, and inhibiting the opposing kinase, Wee1p. We have investigated the ability of Schizosaccharomyces pombe Clp1p, a Cdc14p homolog, to disrupt this auto amplification loop. We show here that Clp1p is required to dephosphorylate, destabilize, and inactivate Cdc25p at the end of mitosis. Clp1p promotes recognition of Cdc25p by the anaphase-promoting complex/cyclosome, an E3 ubiquitin ligase. Failure to inactivate and destabilize Cdc25p in late mitosis delays progression through anaphase, interferes with septation initiation network signaling, and additionally advances the commitment to mitotic entry in the next cycle. This may be a widely conserved mechanism whereby Cdc14 proteins contribute to Cdk1p inactivation. PMID- 14765110 TI - The Yersinia adhesin YadA collagen-binding domain structure is a novel left handed parallel beta-roll. AB - The crystal structure of the recombinant collagen-binding domain of Yersinia adhesin YadA from Yersinia enterocolitica serotype O:3 was solved at 1.55 A resolution. The trimeric structure is composed of head and neck regions, and the collagen binding head region is a novel nine-coiled left-handed parallel beta roll. Before the beta-roll, the polypeptide loops from one monomer to the rest, and after the beta-roll the neck region does the same, making the transition from the globular head region to the narrower stalk domain. This creates an intrinsically stable 'lock nut' structure. The trimeric form of YadA is required for collagen binding, and mutagenesis of its surface residues allowed identification of a putative collagen-binding surface. Furthermore, a new structure-sequence motif for YadA beta-roll was used to identify putative YadA head-like domains in a variety of human and plant pathogens. Such domains may therefore be a common bacterial strategy for avoiding host response. PMID- 14765111 TI - Formation, maintenance and consequences of the imprint at the mating-type locus in fission yeast. AB - Mating-type switching in the fission yeast Schizosaccharomyces pombe is initiated by a strand-specific imprint located at the mating-type (mat1) locus. We show that the imprint corresponds to a single-strand DNA break (SSB), which is site- but not sequence-specific. We identified three novel cis-acting elements, involved in the formation and stability of the SSB. One of these elements is essential for a replication fork pause next to mat1 and interacts in vivo with the Swi1 protein. Another element is essential for maintaining the SSB during cell cycle progression. These results suggest that the DNA break appears during the S-phase and is actively protected against repair. Consequently, during the following round of replication, a polar double-strand break is formed. We show that when the replication fork encounters the SSB, the leading-strand DNA polymerase is able to synthesize DNA to the edge of the SSB, creating a blunt ended recombination intermediate. PMID- 14765112 TI - A troponin switch that regulates muscle contraction by stretch instead of calcium. AB - The flight muscles of many insects have a form of regulation enabling them to contract at high frequencies. The muscles are activated by periodic stretches at low Ca2+ levels. The same muscles also give isometric contractions in response to higher Ca2+. We show that the two activities are controlled by different isoforms of TnC (F1 and F2) within single myofibrils. F1 binds one Ca2+ with high affinity in the C-terminal domain and F2 binds one Ca2+ in the C-terminal domain and one exchangeable Ca2+ in the N-terminal domain. We have characterised the isoforms and determined their effect on the development of stretch-activated and Ca2+ activated tension by replacing endogenous TnC in Lethocerus flight muscle fibres with recombinant isoforms. Fibres with F1 gave stretch-activated tension and minimal isometric tension; those with F2 gave Ca2+-dependent isometric tension and minimal stretch-activated tension. Regulation by a TnC responding to stretch rather than Ca2+ is unprecedented and has resulted in the ability of insect flight muscle to perform oscillatory work at low Ca2+ concentrations, a property to which a large number of flying insects owe their evolutionary success. PMID- 14765113 TI - DIP (mDia interacting protein) is a key molecule regulating Rho and Rac in a Src dependent manner. AB - Cell movement is driven by the coordinated regulation of cytoskeletal reorganization through Rho GTPases downstream of integrin and growth-factor receptor signaling. We have reported that mDia, a target protein of Rho, interacts with Src and DIP. Here we show that DIP binds to p190RhoGAP and Vav2, and that DIP is phosphorylated by Src and mediates the phosphorylation of p190RhoGAP and Vav2 upon EGF stimulation. When endogenous DIP was inhibited by expressing dominant-negative mutants of DIP or siRNA, phosphorylation of p190RhoGAP and Vav2 upon EGF stimulation was diminished, and EGF-induced actin organization, distribution of p190RhoGAP and Vav2, and cell movement were affected. Therefore, DIP seems to transfer the complex of the three proteins from cytosol to beneath the membrane, and the three proteins, in turn, can be phosphorylated by Src. DIP inactivated Rho and activated Rac following EGF stimulation in the membrane fraction. Thus, DIP acts as a regulatory molecule causing Src kinase-dependent feedback modulation of Rho GTPases downstream of Rho mDia upon EGF stimulation, and plays an important role in cell motility. PMID- 14765114 TI - Crystal structure of a myristoylated CAP-23/NAP-22 N-terminal domain complexed with Ca2+/calmodulin. AB - A variety of viral and signal transduction proteins are known to be myristoylated. Although the role of myristoylation in protein-lipid interaction is well established, the involvement of myristoylation in protein-protein interactions is less well understood. CAP-23/NAP-22 is a brain-specific protein kinase C substrate protein that is involved in axon regeneration. Although the protein lacks any canonical calmodulin (CaM)-binding domain, it binds CaM with high affinity. The binding of CAP-23/NAP-22 to CaM is myristoylation dependent and the N-terminal myristoyl group is directly involved in the protein-protein interaction. Here we show the crystal structure of Ca2+-CaM bound to a myristoylated peptide corresponding to the N-terminal domain of CAP-23/NAP-22. The myristoyl moiety of the peptide goes through a hydrophobic tunnel created by the hydrophobic pockets in the N- and C-terminal domains of CaM. In addition to the myristoyl group, several amino-acid residues in the peptide are important for CaM binding. This is a novel mode of binding and is very different from the mechanism of binding in other CaM-target complexes. PMID- 14765115 TI - Molecular architecture of photosynthetic membranes in Rhodobacter sphaeroides: the role of PufX. AB - The effects of the PufX polypeptide on membrane architecture were investigated by comparing the composition and structures of photosynthetic membranes from PufX+ and PufX- strains of Rhodobacter sphaeroides. We show that this single polypeptide profoundly affects membrane morphology, leading to highly elongated cells containing extended tubular membranes. Purified tubular membranes contain helical arrays composed solely of dimeric RC-LH1-PufX (RC, reaction centre; LH, light harvesting) complexes with apparently open LH1 rings. PufX- cells contain crystalline membranes with a pseudo-hexagonal packing of monomeric core complexes. Analysis of purified complexes by electron microscopy and atomic force microscopy shows that LH1 and PufX form a continuous ring of protein around each RC. A model of the tubular membrane is presented with PufX located adjacent to the stained region created by a vacant LH1beta. This arrangement, coupled with a flexible ring, would give the RC QB site transient access to the interstices in the lattice, which might be of functional importance. We discuss the implications of our data for the export of quinol from the RC, for eventual reduction of the cytochrome bc1 complex. PMID- 14765116 TI - In vivo assembly and disassembly of Rad51 and Rad52 complexes during double strand break repair. AB - Assembly and disassembly of Rad51 and Rad52 complexes were monitored by immunofluorescence during homologous recombination initiated by an HO endonuclease-induced double-strand break (DSB) at the MAT locus. DSB-induced Rad51 and Rad52 foci colocalize with a TetR-GFP focus at tetO sequences adjacent to MAT. In strains in which HO cleaves three sites on chromosome III, we observe three distinct foci that colocalize with adjacent GFP chromosome marks. We compared the kinetics of focus formation with recombination intermediates and products when HO-cleaved MATalpha recombines with the donor, MATa. Rad51 assembly occurs 1 h after HO cleavage. Rad51 disassembly occurs at the same time that new DNA synthesis is initiated after single-stranded (ss) MAT DNA invades MATa. We present evidence for three distinct roles for Rad52 in recombination: a presynaptic role necessary for Rad51 assembly, a synaptic role with Rad51 filaments, and a postsynaptic role after Rad51 dissociates. Additional biochemical studies suggest the presence of an ssDNA complex containing both Rad51 and Rad52. PMID- 14765117 TI - Preprotein recognition by the Toc complex. AB - The Toc core complex consists of the pore-forming Toc75 and the GTPases Toc159 and Toc34. We confirm that the receptor form of Toc159 is integrated into the membrane. The association of Toc34 to Toc75/Toc159 is GTP dependent and enhanced by preprotein interaction. The N-terminal half of the pSSU transit peptide interacts with high affinity with Toc159, whereas the C-terminal part stimulates its GTP hydrolysis. The phosphorylated C-terminal peptide of pSSU interacts strongly with Toc34 and therefore inhibits binding and translocation of pSSU into Toc proteoliposomes. In contrast, Toc159 recognises only the dephosphorylated forms. The N-terminal part of the pSSU presequence does not influence binding to the Toc complex, but is able to block import into proteoliposomes through its interaction with Toc159. We developed a model of differential presequence recognition by Toc34 and Toc159. PMID- 14765118 TI - Structural basis of HP1/PXVXL motif peptide interactions and HP1 localisation to heterochromatin. AB - HP1 family proteins are adaptor molecules, containing two related chromo domains that are required for chromatin packaging and gene silencing. Here we present the structure of the chromo shadow domain from mouse HP1beta bound to a peptide containing a consensus PXVXL motif found in many HP1 binding partners. The shadow domain exhibits a novel mode of peptide recognition, where the peptide binds across the dimer interface, sandwiched in a beta-sheet between strands from each monomer. The structure allows us to predict which other shadow domains bind similar PXVXL motif-containing peptides and provides a framework for predicting the sequence specificity of the others. We show that targeting of HP1beta to heterochromatin requires shadow domain interactions with PXVXL-containing proteins in addition to chromo domain recognition of Lys-9-methylated histone H3. Interestingly, it also appears to require the simultaneous recognition of two Lys 9-methylated histone H3 molecules. This finding implies a further complexity to the histone code for regulation of chromatin structure and suggests how binding of HP1 family proteins may lead to its condensation. PMID- 14765119 TI - An extracellular aspartic protease functions in Arabidopsis disease resistance signaling. AB - We have used activation tagging with T-DNA carrying cauliflower mosaic virus 35S enhancers to investigate the complex signaling networks underlying disease resistance in Arabidopsis. From a screen of approximately 5000 lines, we identified constitutive disease resistance (CDR1) encoding an apoplastic aspartic protease, the overexpression of which causes dwarfing and resistance to virulent Pseudomonas syringae. These phenotypes reflect salicylic-acid-dependent activation of micro-oxidative bursts and various defense-related genes. Antisense CDR1 plants were compromised for resistance to avirulent P. syringae and more susceptible to virulent strains than wild type. CDR1 accumulates in intercellular fluid in response to pathogen attacks. Induction of CDR1 generates a small mobile signal, and CDR1 action is blocked by the protease inhibitor pepstatin and by mutations in the protease active sites. We propose that CDR1 mediates a peptide signal system involved in the activation of inducible resistance mechanisms. PMID- 14765120 TI - Runx3 regulates mouse TGF-beta-mediated dendritic cell function and its absence results in airway inflammation. AB - Runx3 transcription factor regulates cell lineage decisions in thymopoiesis and neurogenesis. Here we report that Runx3 knockout (KO) mice develop spontaneous eosinophilic lung inflammation associated with airway remodeling and mucus hypersecretion. Runx3 is specifically expressed in mature dendritic cells (DC) and mediates their response to TGF-beta. In the absence of Runx3, DC become insensitive to TGF-beta-induced maturation inhibition, and TGF-beta-dependent Langerhans cell development is impaired. Maturation of Runx3 KO DC is accelerated and accompanied by increased efficacy to stimulate T cells and aberrant expression of beta2-integrins. Lung alveoli of Runx3 KO mice accumulate DC characteristic of allergic airway inflammation. Taken together, abnormalities in DC function and subset distribution may constitute the primary immune system defect, which leads to the eosinophilic lung inflammation in Runx3 KO mice. These data may help elucidate the molecular mechanisms underlying the pathogenesis of allergic airway inflammation in humans. PMID- 14765121 TI - Sra-1 and Nap1 link Rac to actin assembly driving lamellipodia formation. AB - The Rho-GTPase Rac1 stimulates actin remodelling at the cell periphery by relaying signals to Scar/WAVE proteins leading to activation of Arp2/3-mediated actin polymerization. Scar/WAVE proteins do not interact with Rac1 directly, but instead assemble into multiprotein complexes, which was shown to regulate their activity in vitro. However, little information is available on how these complexes function in vivo. Here we show that the specifically Rac1-associated protein-1 (Sra-1) and Nck-associated protein 1 (Nap1) interact with WAVE2 and Abi 1 (e3B1) in resting cells or upon Rac activation. Consistently, Sra-1, Nap1, WAVE2 and Abi-1 translocated to the tips of membrane protrusions after microinjection of constitutively active Rac. Moreover, removal of Sra-1 or Nap1 by RNA interference abrogated the formation of Rac-dependent lamellipodia induced by growth factor stimulation or aluminium fluoride treatment. Finally, microinjection of an activated Rac failed to restore lamellipodia protrusion in cells lacking either protein. Thus, Sra-1 and Nap1 are constitutive and essential components of a WAVE2- and Abi-1-containing complex linking Rac to site-directed actin assembly. PMID- 14765122 TI - Constitutive versus regulated SNARE assembly: a structural basis. AB - SNARE complex formation is essential for intracellular membrane fusion. Vesicle associated (v-) SNARE intertwines with target membrane (t-) SNARE to form a coiled coil that bridges two membranes and facilitates fusion. For the SNARE family involved in neuronal communications, complex formation is tightly regulated by the v-SNARE-membrane interactions. However, it was found using EPR that complex formation is spontaneous for a different SNARE family that is involved in protein trafficking in yeast. Further, reconstituted yeast SNAREs promoted membrane fusion, different from the inhibited fusion for reconstituted neuronal SNAREs. The EPR structural analysis showed that none of the coiled-coil residues of yeast v-SNARE is buried in the hydrophobic layer of the membrane, making the entire coiled-coil motif accessible, again different from the deep insertion of the membrane-proximal region of neuronal v-SNARE into the bilayer. Importantly, yeast membrane fusion is constitutively active, while synaptic membrane fusion is regulated, consistent with the present results for two SNARE families. Thus, the v-SNARE-membrane interaction may be a major molecular determinant for regulated versus constitutive membrane fusion in cells. PMID- 14765124 TI - DNA topology, not DNA sequence, is a critical determinant for Drosophila ORC-DNA binding. AB - Drosophila origin recognition complex (ORC) localizes to defined positions on chromosomes, and in follicle cells the chorion gene amplification loci are well studied examples. However, the mechanism of specific localization is not known. We have studied the DNA binding of DmORC to investigate the cis-requirements for DmORC:DNA interaction. DmORC displays at best six-fold differences in the relative affinities to DNA from the third chorion locus and to random fragments in vitro, and chemical probing and DNase1 protection experiments did not identify a discrete binding site for DmORC on any of these fragments. The intrinsic DNA binding specificity of DmORC is therefore insufficient to target DmORC to origins of replication in vivo. However, the topological state of the DNA significantly influences the affinity of DmORC to DNA. We found that the affinity of DmORC for negatively supercoiled DNA is about 30-fold higher than for either relaxed or linear DNA. These data provide biochemical evidence for the notion that origin specification in metazoa likely involves mechanisms other than simple replicator initiator interactions and that in vivo other proteins must determine ORC's localization. PMID- 14765123 TI - DCX, a new mediator of the JNK pathway. AB - Mutations in the X-linked gene DCX result in lissencephaly in males, and abnormal neuronal positioning in females, suggesting a role for this gene product during neuronal migration. In spite of several known protein interactions, the involvement of DCX in a signaling pathway is still elusive. Here we demonstrate that DCX is a substrate of JNK and interacts with both c-Jun N-terminal kinase (JNK) and JNK interacting protein (JIP). The localization of this signaling module in the developing brain suggests its functionality in migrating neurons. The localization of DCX at neurite tips is determined by its interaction with JIP and by the interaction of the latter with kinesin. DCX is phosphorylated by JNK in growth cones. DCX mutated in sites phosphorylated by JNK affected neurite outgrowth, and the velocity and relative pause time of migrating neurons. We hypothesize that during neuronal migration, there is a need to regulate molecular motors that are working in the cell in opposite directions: kinesin (a plus-end directed molecular motor) versus dynein (a minus-end directed molecular motor). PMID- 14765125 TI - Nrdp1-mediated degradation of the gigantic IAP, BRUCE, is a novel pathway for triggering apoptosis. AB - Degradation of certain inhibitor of apoptosis proteins (IAPs) appears to be critical in the initiation of apoptosis, but the factors that regulate their degradation in mammalian cells are unknown. Nrdp1/FLRF is a RING finger containing ubiquitin ligase that catalyzes degradation of the EGF receptor family member, ErbB3. We show here that Nrdp1 associates with BRUCE/apollon, a 530 kDa membrane-associated IAP, which contains a ubiquitin-carrier protein (E2) domain. In the presence of an exogenous E2, UbcH5c, purified Nrdp1 catalyzes BRUCE ubiquitination. In vivo, overexpression of Nrdp1 promotes ubiquitination and proteasomal degradation of BRUCE. In many cell types, apoptotic stimuli induce proteasomal degradation of BRUCE (but not of XIAP or c-IAP1), and decreasing Nrdp1 levels by RNA interference reduces this loss of BRUCE. Furthermore, decreasing BRUCE content by RNA interference or overexpression of Nrdp1 promotes apoptosis. Thus, BRUCE normally inhibits apoptosis, and Nrdp1 can be important in the initiation of apoptosis by catalyzing ubiquitination and degradation of BRUCE. PMID- 14765126 TI - A Suv39h-dependent mechanism for silencing S-phase genes in differentiating but not in cycling cells. AB - The Rb/E2F complex represses S-phase genes both in cycling cells and in cells that have permanently exited from the cell cycle and entered a terminal differentiation pathway. Here we show that S-phase gene repression, which involves histone-modifying enzymes, occurs through distinct mechanisms in these two situations. We used chromatin immunoprecipitation to show that methylation of histone H3 lysine 9 (H3K9) occurs at several Rb/E2F target promoters in differentiating cells but not in cycling cells. Furthermore, phenotypic knock down experiments using siRNAs showed that the histone methyltransferase Suv39h is required for histone H3K9 methylation and subsequent repression of S-phase gene promoters in differentiating cells, but not in cycling cells. These results indicate that the E2F target gene permanent silencing mechanism that is triggered upon terminal differentiation is distinct from the transient repression mechanism in cycling cells. Finally, Suv39h-depleted myoblasts were unable to express early or late muscle differentiation markers. Thus, appropriately timed H3K9 methylation by Suv39h seems to be part of the control switch for exiting the cell cycle and entering differentiation. PMID- 14765127 TI - Tyrosine phosphorylation controls Runx2-mediated subnuclear targeting of YAP to repress transcription. AB - Src/Yes tyrosine kinase signaling contributes to the regulation of bone homeostasis and inhibits osteoblast activity. Here we show that the endogenous Yes-associated protein (YAP), a mediator of Src/Yes signaling, interacts with the native Runx2 protein, an osteoblast-related transcription factor, and suppresses Runx2 transcriptional activity in a dose-dependent manner. Runx2, through its PY motif, recruits YAP to subnuclear domains in situ and to the osteocalcin (OC) gene promoter in vivo. Inhibition of Src/Yes kinase blocks tyrosine phosphorylation of YAP and dissociates endogenous Runx2-YAP complexes. Consequently, recruitment of the YAP co-repressor to subnuclear domains is abrogated and expression of the endogenous OC gene is induced. Our results suggest that Src/Yes signals are integrated through organization of Runx2-YAP transcriptional complexes at subnuclear sites to attenuate skeletal gene expression. PMID- 14765128 TI - The phagocyte NADPH oxidase depends on cholesterol-enriched membrane microdomains for assembly. AB - The superoxide-producing phagocyte NADPH oxidase consists of a membrane-bound flavocytochrome b558 complex, and cytosolic factors p47phox, p67phox and the small GTPase Rac, which translocate to the membrane to assemble the active complex following cell activation. We here show that insolubility of NADPH oxidase subunits in nonionic detergents TX-100, Brij-58, and Brij-98 is a consequence of inclusion into cholesterol-enriched membrane microdomains (lipid rafts). Thus, flavocytochrome b558, in a cholesterol-dependent manner, segregated to the bouyant low-density detergent-resistant membrane (DRM) fraction, and the cytosolic NADPH oxidase factors associated dynamically with low-density DRM. Further, superoxide production following cholesterol depletion was severely compromised in intact cells or in a cell-free reconstituted system, correlating with a reduced translocation of cytosolic phox subunits to the membrane. In analogy with the widely accepted role of lipid rafts as signaling platforms, our data indicate that cholesterol-enriched microdomains act to recruit and/or organize the cytosolic NADPH oxidase factors in the assembly of the active NADPH oxidase. PMID- 14765130 TI - Adenoviral-mediated PTEN transgene expression sensitizes Bcl-2-expressing prostate cancer cells to radiation. AB - Bcl-2 is associated with resistance to radiotherapy in prostate cancer. It was recently demonstrated that transduction of LNCaP prostate cells with the PTEN gene resulted in Bcl-2 downregulation. We hypothesized that forced expression of PTEN in prostate cancer cells would sensitize cells to radiation, downregulate Bcl-2 expression, and potentiate the G2M block induced by radiation. Four cell lines - PC-3-Bcl-2 (Bcl-2 overexpression, deleted PTEN), PC-3-Neo (wild-type Bcl 2, deleted PTEN), LNCaP (Bcl-2 overexpression, deleted PTEN), and DU-145 (wild type Bcl-2 and PTEN) - were transduced with a recombinant adenovirus-5 vector expressing the human wild-type PTEN cDNA under the control of a human cytomegalovirus promoter (Ad-MMAC). After correction for the effect of Ad-MMAC on plating efficiency, Ad-MMAC treatment reduced the surviving fractions after 2 Gy as follows: PC-3-Bcl-2, from 60.5 to 3.6%; PC-3-Neo, no reduction; LNCaP, from 29.6 to 16.3%; and DU-145, from 32.7 to 25.7%. PTEN expression was associated with the downregulation of Bcl-2 expression in PC-3-Bcl-2 and LNCaP cell lines. Ad-MMAC plus radiotherapy potentiated the G2M block seen with radiotherapy alone only in PC-3-Bcl-2 cells. These findings suggest that overexpression of Bcl-2 result in radioresistance and inability of radiation to cause its typical G2M cell-cycle arrest. PMID- 14765129 TI - An RNA-dependent protein kinase is involved in tunicamycin-induced apoptosis and Alzheimer's disease. AB - Various types of stress, such as disruption of calcium homeostasis, inhibition of protein glycosylation and reduction of disulfide bonds, result in accumulation of misfolded proteins in the endoplasmic reticulum (ER). The initial cellular response involves removal of such proteins by the ER, but excessive and/or long term stress results in apoptosis. In this study, we used a randomized ribozyme library and ER stress-mediated apoptosis (tunicamycin-induced apoptosis) in SK-N SH human neuroblastoma cells as a selective phenotype to identify factors involved in this process. We identified a double-stranded RNA-dependent protein kinase (PKR) as one of the participants in this process. The level of nuclear PKR was elevated, but the level of cytoplasmic PKR barely changed in tunicamycin treated SK-N-SH cells. Furthermore, tunicamycin also raised levels of phosphorylated PKR in the nucleus. We also detected the accumulation of phosphorylated PKR in the nuclei of autopsied brain tissues in Alzheimer's disease. Thus, PKR might play a role in ER stress-induced apoptosis and in Alzheimer's disease. PMID- 14765131 TI - Gene therapy for esophageal carcinoma: the use of an explant model to test adenoviral vectors ex vivo. AB - Adenoviral gene therapy might be a promising therapeutic strategy for esophageal carcinoma. However, adenoviral transduction efficacy in vivo is still limited. This efficacy can be improved by the insertion of an Arg-Gly-Asp (RGD) peptide in the HI-loop of the viral fiber knob. Indeed in established esophageal cell lines, we observed an up to six-fold improved transduction using the RGD-targeted adenovirus. Established cell lines, however, are easily transformed and do not represent the more complex in vivo histology and anatomy. Therefore, we set up an esophageal explant model using esophageal biopsies from patients. Viability is a limiting factor for this system. Cultured squamous epithelium, intestinal metaplasia and squamous cell carcinoma had a sufficient viability to study adenoviral transduction. Viability of the cultured adenocarcinoma biopsies was poor. Adenoviral transduction in the explant model was poor and was localized in particular cells. The transduction of the nontargeted and RGD-targeted adenovirus was similar in localization and efficacy. In conclusion, we established an esophageal explant system to test the transduction of adenoviral vectors ex vivo. The transduction was limited and localized in specific cells. RGD-targeted adenovirus did not show an improved transduction in this system. PMID- 14765132 TI - Coupling endoplasmic reticulum stress to the cell death program. AB - The endoplasmic reticulum (ER) regulates protein synthesis, protein folding and trafficking, cellular responses to stress and intracellular calcium (Ca(2+)) levels. Alterations in Ca(2+) homeostasis and accumulation of misfolded proteins in the ER cause ER stress that ultimately leads to apoptosis. Prolonged ER stress is linked to the pathogenesis of several different neurodegenerative disorders. Apoptosis is a form of cell death that involves the concerted action of a number of intracellular signaling pathways including members of the caspase family of cysteine proteases. The two main apoptotic pathways, the death receptor ('extrinsic') and mitochondrial ('intrinsic') pathways, are activated by caspase 8 and -9, respectively, both of which are found in the cytoplasm. Recent studies point to the ER as a third subcellular compartment implicated in apoptotic execution. Here, we review evidence for the contribution of various cellular molecules that contribute to ER stress and subsequent cellular death. It is hoped that dissection of the molecular components and pathways that alter ER structure and function and ultimately promote cellular death will provide a framework for understanding degenerative disorders that feature misfolded proteins. PMID- 14765133 TI - Hydrogen peroxide produced by Aplysia ink toxin kills tumor cells independent of apoptosis via peroxiredoxin I sensitive pathways. AB - Marine snails of the genus Aplysia possess numerous bioactive substances. We have purified a 60 kDa protein, APIT (Aplysia punctata ink toxin), from the defensive ink of A. punctata that triggers cell death with profound tumor specificity. Tumor cell death induced by APIT is independent of apoptosis but is characterized by the rapid loss of metabolic activity, membrane permeabilization, and shrinkage of nuclei. Proteome analysis of APIT-treated tumor cells indicated a modification of peroxiredoxin I, a cytoplasmic peroxidase involved in the detoxification of peroxides. Interestingly, knockdown of peroxiredoxin I expression by RNA interference sensitized cells for APIT-induced cell death. APIT induced the death of tumor cells via the enzymatic production of H2O2 and catalase completely blocked APITs' activity. Our data suggest that H2O2 induced stress and the modulation of peroxiredoxins might be a promising approach for tumor therapy. PMID- 14765134 TI - N-(4-hydroxyphenyl) retinamide (4HPR) enhances TRAIL-mediated apoptosis through enhancement of a mitochondrial-dependent amplification loop in ovarian cancer cell lines. AB - The majority of ovarian cancer cells are resistant to apoptosis induced by tumor necrosis factor-related apoptosis-inducing ligand (TRAIL). Subtoxic concentrations of the semisynthetic retinoid N-(4-hydroxyphenyl)retinamide (4HPR) enhanced TRAIL-mediated apoptosis in ovarian cancer cell lines but not in immortalized nontumorigenic ovarian epithelial cells. The enhancement of TRAIL mediated apoptosis by 4HPR was not due to changes in the levels of proteins known to modulate TRAIL sensitivity. The combination of 4HPR and TRAIL enhanced cleavage of multiple caspases in the death receptor pathway (including the two initiator caspases, caspase-8 and caspase-9). The 4HPR and TRAIL combination leads to mitochondrial permeability transition, significant increase in cytochrome c release, and increased caspase-9 activation. Caspase-9 may further activate caspase-8, generating an amplification loop. Stable overexpression of Bcl-xL abrogates the interaction between 4HPR and TRAIL at the mitochondrial level by blocking cytochrome c release. As a consequence, a decrease in activation of caspase-9, caspase-8, and TRAIL-mediated apoptosis occurs. These results indicate that the enhancement in TRAIL-mediated apoptosis induced by 4HPR is due to the increase in activation of multiple caspases involving an amplification loop via the mitochondrial-death pathway. These findings offer a promising and novel strategy for the treatment of ovarian cancer. PMID- 14765135 TI - Human death effector domain-associated factor interacts with the viral apoptosis agonist Apoptin and exerts tumor-preferential cell killing. AB - Apoptin, a protein from chicken anemia virus without an apparent cellular homologue, can induce apoptosis in mammalian cells. Its cytotoxicity is limited to transformed or tumor cells, making Apoptin a highly interesting candidate for cancer therapy. To elucidate Apoptin's mechanism of action, we have searched for binding partners in the human proteome. Here, we report that Apoptin interacts with DEDAF, a protein previously found to associate with death effector domain (DED)-containing pro-apoptotic proteins, and to be involved in regulation of transcription. Like Apoptin, after transient overexpression, DEDAF induced apoptosis in various human tumor cell lines, but not in primary fibroblasts or mesenchymal cells. DEDAF-induced cell death was inhibited by the caspase inhibitor p35. Together with the reported association of DEDAF with a DED containing DNA-binding protein in the nucleus and the transcription regulatory activity, our findings may provide a clue for the mechanism of Apoptin's actions in mammalian cells. PMID- 14765136 TI - RAIDD aggregation facilitates apoptotic death of PC12 cells and sympathetic neurons. AB - In human cell lines, the caspase 2 adaptor RAIDD interacts selectively with caspase 2 through its caspase recruitment domain (CARD) and leads to caspase 2 dependent death. Whether RAIDD induces such effects in neuronal cells is unknown. We have previously shown that caspase 2 is essential for apoptosis of trophic factor-deprived PC12 cells and rat sympathetic neurons. We report here that rat RAIDD, cloned from PC12 cells, interacts with rat caspase 2 CARD. RAIDD overexpression induced caspase 2 CARD- and caspase 9-dependent apoptosis of PC12 cells and sympathetic neurons. Apoptosis correlated with the formation of discrete perinuclear aggregates. Both death and aggregates required the expression of full-length RAIDD. Such aggregates may enable more effective activation of caspase 2 through close proximity. Following trophic deprivation, RAIDD overexpression increased death and aggregate formation. Therefore, RAIDD aggregation is important for its death-promoting effects and may play a role in trophic factor withdrawal-induced neuronal apoptosis. PMID- 14765137 TI - Noninvasive assessment of the injured human spinal cord by means of functional magnetic resonance imaging. AB - STUDY DESIGN: A magnetic resonance imaging technique that enables indirect detection of neuronal activity has been developed for the spinal cord. In the present study, this method, spinal functional magnetic resonance imaging (fMRI), is applied to the first study of the injured spinal cord, with the goal of better clinical assessment of the entire cord. OBJECTIVES: The objectives of this project are: (1) to investigate the neuronal activity that can be detected in the spinal cord caudal to a chronic injury by means of spinal fMRI, and (2) to develop spinal fMRI as a clinical diagnostic tool. SETTING: Institute for Biodiagnostics, National Research Council of Canada, Winnipeg, Manitoba, Canada. METHODS: fMRI of the spinal cord was carried out in 27 volunteers with cervical or thoracic spinal cord injuries (SCIs). Of these volunteers, 18 had complete injuries, and nine had incomplete injuries. Spinal fMRI was carried out in a 1.5 T clinical MR system, using established methods. Thermal stimulation at 10 degrees C was applied to the fourth lumbar dermatome on each leg, and images were obtained of the entire lumbar spinal cord. RESULTS: Areas of neuronal activity were consistently observed in the lumbar spinal cord in response to the thermal stimulation, even when the subjects had no awareness of the sensation. The pattern of activity was notably different compared with noninjured subjects. In general, subjects with complete SCI showed absent or diminished dorsal gray matter activity, but had enhanced ventral activity, particularly contralateral to the stimulation. CONCLUSIONS: Spinal fMRI is able to provide a noninvasive assessment of the injured spinal cord that does not depend on the patient's perception of the stimulus being applied. This work was carried out on a standard clinical MRI system without modification, and so is readily applicable in most MR units. SPONSORSHIP: This work was funded by a grant from the Canadian Institutes of Health Research (CIHR). PMID- 14765138 TI - Lumbar repetitive magnetic stimulation reduces spastic tone increase of the lower limbs. AB - STUDY DESIGN: Comparison of spinal lesion subjects and normal subjects. OBJECTIVE: To investigate the effects of a paravertebral repetitive magnetic stimulation on spastic tone increase of the lower limbs. SETTING: Munich, Germany. METHODS: We compared the effects in 15 patients with different spinal lesions and in 16 healthy subjects. The spastic tone increase was evaluated clinically with the Ashworth scale and apparatively with the pendulum test, both at fixed times before and after stimulation. Unilateral stimulation was applied to the lumbar nerve roots L3 and L4 of the clinically more spastic leg. RESULTS: The spastic tone decreased significantly in the interval between 4 and 24 h after stimulation. This effect was slightly more pronounced in the contralateral extremity. Furthermore, the stimulation motor threshold of the patients was significantly raised. CONCLUSION: Repetitive magnetic unilateral stimulation has a positive effect on spastic tone increase due to spinal lesions, causing a decrease that lasts for about 1 day not only on the ipsilateral but also on the contralateral side. PMID- 14765139 TI - Levels of brain wave activity (8-13 Hz) in persons with spinal cord injury. AB - STUDY DESIGN: Brain wave activity in people with spinal cord injury (SCI) was compared to brain wave activity in able-bodied controls. OBJECTIVES: To investigate whether a spinal injury results in changes in levels of brain wave activity in the 8-13 Hz spectrum of the electroencephalography (EEG). SETTING: Sydney, Australia. METHODS: Monopolar, multichannel EEG assessment was completed for 20 persons with SCI and 20 able-bodied, sex- and age-matched controls. A total of 14 channels of EEG were measured across the entire scalp for all participants. Comparisons between the able-bodied and SCI participants were made across the frontal, central, parietal, occipital and temporal regions. Comparisons were also made for impairment level, that is, between participants with tetraplegia and paraplegia. RESULTS: Compared to the able-bodied controls, consistently reduced brain wave activity (measured by magnitude and peak amplitude) in the 8-13 Hz component of the EEG occurred in persons with SCI across all regions and sites, and differences were larger in the central, parietal and occipital sites. The SCI group also had consistently lower frequencies than the able-bodied controls. Furthermore, the subgroup of SCI participants with tetraplegia generally had significantly reduced brain activity (magnitude and peak amplitude) compared with the paraplegic subgroup and able bodied controls. CONCLUSIONS: The findings of this research enhance our understanding of changes in brain wave activity that could be associated with deafferentation that occurs following SCI, as well as provide essential data on the potential of SCI persons to use a 'hands free' environmental control system that is based upon 8-13 Hz brain activity. PMID- 14765140 TI - Motor capacities of upper limbs in tetraplegics: a new scale for the assessment of the results of functional surgery on upper limbs. AB - STUDY DESIGN: Metrological investigation. OBJECTIVES: To study the validity and the reliability of a Motor Capacities Scale (MCS) specifically designed for tetraplegics who undergo a functional surgery of upper limbs. SETTING: Bouffard Vercelli Centre, Cerbere, France. METHODS: From diverse sources (observation of patients, review of literature, discussions with occupational therapists and physicians), we compiled a list of 300 activities relating to daily living tasks. From this list, 80 items of motor capacities were retained. They correspond to the abilities of a patient to perform basic and functional tasks regardless of contextual factors (environmental and personal factors). In a preliminary study, items were reviewed by occupational therapists and by 40 tetraplegics for criticism. An open study and an intermediate study were conducted to assess, respectively, the feasibility and the reliability of the MCS. A prefinal study was focused on construct validity. Convergent and divergent hypotheses were formulated and tested against other measures. Four criteria were identified for the elimination of some of the 36 remaining items: a variance of the item equal to 0, a low reproducibility, a high level of redundancy studied by item-to-item correlation and a low level of comprehension. RESULTS: In all, 52 tetraplegics were included in the prefinal study. Global inter-rater reproducibility was excellent (intraclass correlation coefficient of 0.99). In accordance with the criteria of elimination, the number of items was reduced to 31. Correlations with the Sollerman test and the Asia Motor Score were, respectively, 0.959 and 0.7444 (P<0.0001). Correlation with the interval since the onset of the tetraplegia and the educational level were, respectively, 0.20 and 0.195 (P=0.163). CONCLUSION: MCS displays a good apparent and content validity, and excellent reproducibility and construct validity. Metrological properties were good enough to allow the evaluation of sensitivity to change in the final study. PMID- 14765141 TI - Submaximal physical strain and peak performance in handcycling versus handrim wheelchair propulsion. AB - STUDY DESIGN: Experimental study in subjects with paraplegia and nondisabled subjects. OBJECTIVE: To compare submaximal physical strain and peak performance in handcycling and handrim wheelchair propulsion in wheelchair-dependent and nondisabled control subjects SETTING: Amsterdam, The Netherlands. METHODS: Nine male subjects with paraplegia and 10 nondisabled male subjects performed two exercise tests on a motor-driven treadmill using a handrim wheelchair and attach unit handcycle system. The exercise protocol consisted of two 4-min submaximal exercise bouts at 25 and 35 W, followed by 1-min exercise bouts with increasing power output until exhaustion. RESULTS: Analysis of variance for repeated measures showed a significantly lower oxygen uptake (VO2), ventilation (Ve), heart rate (HR), rate of perceived exertion and a higher gross efficiency for handcycling at 35 W in both subject groups, while no significant differences were found at 25 W. Peak power output and peak VO2, Ve and HR were significantly higher during handcycling in both groups. The differences between handcycling and wheelchair propulsion were the same in subjects with paraplegia and the nondisabled subjects. CONCLUSIONS: Handcycling induces significantly less strain at a moderate submaximal level of 35 W, and shows noticeably higher maximal exercise responses than wheelchair propulsion, which is consistent in subjects with paraplegia and nondisabled controls. These results demonstrate that handcycling is beneficial for mobility in daily life of wheelchair users. PMID- 14765142 TI - Characteristics of radiogenic lower motor neurone disease, a possible link with a preceding viral infection. AB - OBJECTIVE: To investigate the pathogenesis of the rare radiogenic lower motor neurone disease (LMND) on the basis of a meta-analysis of the published case histories. MATERIALS AND METHODS: The authors reviewed 47 well-documented radiogenic LMND cases from the English literature. RESULTS: The disease typically occurs following the irradiation of radiosensitive cancers situated near the spinal cord. It arises predominantly (46 cases) in the lower extremities; only one case involved the upper extremities. There is a male predominance (male:female ratio 7.8:1), and the patients are characteristically young (13-40 years, with four exceptions). An overdose does not seem to be a particular risk factor for the development of the disease, as total dose, fraction size and biologically effective dose are typically below 50 Gy, 2 Gy and 128 Gy2, respectively, which are regarded as safe doses. Other risk factors (chemotherapy, operations, etc) have been identified only rarely. Radiogenic LMND is manifested in an apparently random manner, 4-312 (mean 48.7) months after the completion of radiotherapy. DISCUSSION: The complete lack of a dose-effect relationship argues strongly against a pure radiogenic nature of the pathological process. The latency period is typically several years and it varies extremely, which excludes a direct and complete causal relationship between radiotherapy and LMND. As the interaction of ionizing radiation with living tissues is highly unspecific, thus a selective motor injury due to irradiation alone, without comparable effects on the sensory and vegetative fibers, seems improbable. CONCLUSIONS: On analogy with the viral motor neurone diseases, we suppose that radiogenic LMND may be preceded by viral (enterovirus/poliovirus) infection. Based on the meta-analysis, it is suggested that irradiation may be only a single component of the set of factors jointly resulting in the clinical state regarded as radiogenic LMND. PMID- 14765143 TI - Urethral cultures in patients with spinal cord injury. AB - STUDY DESIGN: Prospective analysis. OBJECTIVES: To investigate the colonization of the distal urethra and bladder during the initial stages of rehabilitation in acute spinal-cord injury (SCI) and to examine the association between bacteriuria and colonization of the distal urethra. SETTING: Selcuk University Meram Faculty of Medicine, Konya, Turkey. METHODS: A total of 27 patients with SCI (13 females and 14 males) and 40 controls without evidence of disease of the urinary tract were studied. Cultures were taken from the patients who applied clean intermittent catheterization and compared with normal subjects. RESULTS: Escherichia coli was predominantly isolated from the urine and urethral cultures of both female and male SCI patients. Colonization of other bacteria in the urine and urethral cultures was similar in both female and male patients, except for Pseudomonas, which was colonized in male patients. In all, 72% of patients who had E. coli positive urethral cultures also had E. coli colonization in their simultaneous urine cultures. There was concordance between urethra and urine cultures concerning the growth of E. coli (P=0.82). When urethral cultures collected 1 week before were evaluated in patients with E. coli positive urine cultures (n=24 cultures), 15 of these urethral cultures also had E. coli colonization. There was concordance between urethra and urine cultures concerning the growth of E. coli (P=0.66). CONCLUSIONS: Our study suggested that urethral flora was a significant source for the development of urinary infection in spinal cord-injured patients. PMID- 14765144 TI - Bladder-filling sensation in patients with spinal cord injury and the potential for sensation-dependent bladder emptying. AB - STUDY DESIGN: Investigation of bladder-filling sensation in 73 consecutive traumatic spinal cord injury (SCI) patients during laboratory cystometry. OBJECTIVES: To determine the frequencies of bladder-filling sensation in SCI patients with complete lesions above T11 and below T10 and with incomplete lesions, to examine the quality of the preserved sensation, as well as to determine the potential for sensation-dependent bladder emptying in this patient group. SETTING: Physical Medicine and Rehabilitation Education and Research Hospital of Ministry of Health, Ankara, Turkey. METHODS: Bladder-filling sensation was investigated in 73 consecutive patients with traumatic SCI using continuous medium fill cystometry. Bladder-filling sensation was categorised as absent, partially preserved or preserved. Presence of bladder-filling sensation before intravesical pressure (Pves) reached 25 and 40 cmH2O, and simultaneous presence of bladder capacity of more than 150 ml were investigated as the measures of the quality of the sensation. RESULTS: Bladder-filling sensation was present to some degree in all incomplete SCI patients, in 82.4% of the patients with complete lesions below T10, and 38.9% of the patients with complete lesions above T11. There were statistically significant differences between three groups with respect to bladder sensation category (P<0.001). About 86% of the patients with incomplete lesions, 53% of the patients with complete lesions below T10 and 22% of those with lesions above T11 had bladder-filling sensation before Pves reached 25 cmH2O and simultaneous bladder capacity of more than 150 ml was present in 61.2, 41.2 and 22.2% of the patients in the groups, respectively. Bladder-filling sensation investigations were reliable in terms of bladder filling sensation category in 36 SCI patients who had second cystometric examination. CONCLUSION: Presence of bladder-filling sensation in many of the SCI patients observed in this study revealed the potential for sensation-dependent bladder emptying in SCI patients, especially in the ones with complete lesions below T10 and the ones with incomplete lesions. This method may prevent early emptying attempts, unnecessary catheterisations and overdistension episodes and may improve the patient's self-esteem and quality of life. However, further studies on clinical experience and patient compliance for this method are needed. PMID- 14765145 TI - Spinal cord compression in beta-thalassemia: case report and review of the literature. AB - STUDY DESIGN: A case report of thoracic spinal cord compression in a 34-year-old male with beta-thalassemia is reported. OBJECTIVES: In patients with thalassemia, neurologic complaints should lead to a high index of suspicion for spinal cord compression from marrow expansion, ectopic bone formation and resultant stenosis. Initial presentation, diagnosis, radiographic findings, surgical treatment and follow-up are reviewed. SETTING: This case is reported from Chicago, Illinois. METHOD: A chart review is performed for the purposes of this case report. RESULTS: Patient underwent decompressive laminectomy with good surgical outcome. CONCLUSION: Rapid diagnosis and treatment of such a condition is essential to optimize the chances of recovery. PMID- 14765146 TI - Painful Schmorl's node treated by lumbar interbody fusion. AB - STUDY DESIGN: A case report of painful lumbar Schmorl's node is presented. OBJECTIVE: To describe diagnostic evidence and the result of surgical treatment of a rare case of painful Schmorl's node. SETTING: Niigata, Japan. CASE REPORT: A 55-year-old housewife was diagnosed with painful Schmorl's node of L3 by discography, which depicted leakage of the contrast medium into the L3 vertebra through a disruption of the central part of the cranial end plate with concomitant back pain. Segmental fusion surgery was performed. Mechanical low back pain of the patient improved just after surgery. Histologic examination demonstrated that fibrocartilaginous tissue herniated through a disruption of the superior end plate and forced into the vertebral spongiosa. CONCLUSIONS: Painful Schmorl's node can be diagnosed by discography, which demonstrates an intravertebral disc herniation with concomitant back pain. Surgical treatment should be considered in a patient with persistent disabling back pain. When surgical treatment is indicated, eradication of the intervertebral disc including Schmorl's node and segmental fusion are preferable. PMID- 14765147 TI - Abnormal bony protuberance of anterior atlas causing dysphagia. A rare congenital anomaly. AB - STUDY DESIGN: A Case report. OBJECTIVE: To report an exceedingly rare case of dysphagia caused by abnormal bony protuberance of anterior atlas. SETTING: Kocaeli University Faculty of Medicine, Departments of Neurosurgery and Otorhinolaryngology. METHOD: Radiological examinations revealed a large anterior abnormal bony tuberance of atlas in an 11-year-old boy who complained of dysphagia of 5 years donation. RESULT: The anterior bony tuberance of the atlas was resected by a transoral approach. Histopathological examination of the surgical specimens showed normal bone tissue. Resection resulted in complete resolution of the dysphagia. CONCLUSIONS: Dysphagia can be caused by disorders of the cervical spine. These disorders are usually seen in elderly adults. In the pediatric population, spinal abnormality is an exceedingly rare cause of dysphagia. Abnormal inductive signals from the adjacent notocord and ventral neural tube may play role in the pathogenesis of this abnormal bony protuberance. PMID- 14765148 TI - Awareness and use of advance directives in the spinal cord injured population (Spinal Cord 2002; 40: 581-594). PMID- 14765150 TI - Management of recurrent priapism in a cervical spinal cord injury patient with oral baclofen therapy. PMID- 14765151 TI - Complications of sacral anterior root stimulator implantation in a cervical spinal cord injury patient: increased spasms requiring intrathecal baclofen therapy followed by delayed fracture of lumbar spine leading to intractable spasms compelling disuse of the sacral anterior root stimulator. PMID- 14765153 TI - Defeated but not deterred. PMID- 14765154 TI - Sound thinking. PMID- 14765155 TI - Fear of human pandemic grows as bird flu sweeps through Asia. PMID- 14765156 TI - Drug suicide risks prompt call for FDA action. PMID- 14765157 TI - Europe urged to move on transgenic crop imports. PMID- 14765158 TI - Bush's belt-tightening budget offers science slim pickings. PMID- 14765160 TI - Suicide-inquiry fallout 'could gag' scientists. PMID- 14765161 TI - Fusion meeting shelved as site decision slides. PMID- 14765162 TI - Universities battle for extra funds in bid to boost quality. PMID- 14765163 TI - Extinction meeting kicks off Japan's plans for networking. PMID- 14765165 TI - Noise management: sound and vision. PMID- 14765166 TI - Quantum gravity: back to the future. PMID- 14765167 TI - Fertilizer 'solution' could turn local problem global. PMID- 14765168 TI - Framework reveals lack of basic-research funds. PMID- 14765169 TI - Working hard for the money. PMID- 14765175 TI - Prisoners of the dilemma. PMID- 14765177 TI - Planetary science: double trouble. PMID- 14765176 TI - Medicine: K is for koagulation. PMID- 14765178 TI - Cell division: guardian spirit blesses meiosis. PMID- 14765179 TI - Nanomaterials: jagged edge. PMID- 14765180 TI - Inorganic chemistry: ammonia transformed. PMID- 14765181 TI - Ion channels: shake, rattle or roll? PMID- 14765185 TI - Stress propagation: getting to the bottom of a granular medium. AB - Penetration by an object through a dense granular medium (for example, by a finger pushing slowly into the sand on a beach) presents an interesting physics problem that is closely related to issues of practical importance in soil science. Here we measure the penetration-resistance force for an object approaching the solid bottom boundary of a granular sample--analogous to the finger approaching a flat rock buried in the beach. We find that the penetration resistance near the boundary increases exponentially, which demonstrates the existence of an intrinsic length scale to the 'jamming' caused by a locally applied stress. PMID- 14765186 TI - Transgenic mice: fat-1 mice convert n-6 to n-3 fatty acids. AB - Mammals cannot naturally produce omega-3 (n-3) fatty acids--beneficial nutrients found mainly in fish oil--from the more abundant omega-6 (n-6) fatty acids and so they must rely on a dietary supply. Here we show that mice engineered to carry a fat-1 gene from the roundworm Caenorhabditis elegans can add a double bond into an unsaturated fatty-acid hydrocarbon chain and convert n-6 to n-3 fatty acids. This results in an abundance of n-3 and a reduction in n-6 fatty acids in the organs and tissues of these mice, in the absence of dietary n-3. As well as presenting an opportunity to investigate the roles played by n-3 fatty acids in the body, our discovery indicates that this technology might be adapted to enrich n-3 fatty acids in animal products such as meat, milk and eggs. PMID- 14765187 TI - Mixing, volatile loss and compositional change during impact-driven accretion of the Earth. AB - The degree to which efficient mixing of new material or losses of earlier accreted material to space characterize the growth of Earth-like planets is poorly constrained and probably changed with time. These processes can be studied by parallel modelling of data from different radiogenic isotope systems. The tungsten isotope composition of the silicate Earth yields a model timescale for accretion that is faster than current estimates based on terrestrial lead and xenon isotope data and strontium, tungsten and lead data for lunar samples. A probable explanation for this is that impacting core material did not always mix efficiently with the silicate portions of the Earth before being added to the Earth's core. Furthermore, tungsten and strontium isotope compositions of lunar samples provide evidence that the Moon-forming impacting protoplanet Theia was probably more like Mars, with a volatile-rich, oxidized mantle. Impact-driven erosion was probably a significant contributor to the variations in moderately volatile element abundance and oxidation found among the terrestrial planets. PMID- 14765188 TI - The formation of Kuiper-belt binaries through exchange reactions. AB - Recent observations have revealed that an unexpectedly high fraction--a few per cent--of the trans-Neptunian objects (TNOs) that inhabit the Kuiper belt are binaries. The components have roughly equal masses, with very eccentric orbits that are wider than a hundred times the radius of the primary. Standard theories of binary asteroid formation tend to produce close binaries with circular orbits, so two models have been proposed to explain the unique characteristics of the TNOs. Both models, however, require extreme assumptions regarding the size distribution of the TNOs. Here we report a mechanism that is capable of producing binary TNOs with the observed properties during the early stages of their formation and growth. The only required assumption is that the TNOs were initially formed through gravitational instabilities in the protoplanetary dust disk. The basis of the mechanism is an exchange reaction in which a binary whose primary component is much more massive than the secondary interacts with a third body, whose mass is comparable to that of the primary. The low-mass secondary component is ejected and replaced by the third body in a wide but eccentric orbit. PMID- 14765189 TI - Statistical mechanics of a gas-fluidized particle. AB - Characterization of the microscopic fluctuations in systems that are far from equilibrium is crucial for understanding the macroscopic response. One approach is to use an 'effective temperature'--such a quantity has been invoked for chaotic fluids, spin glasses, glasses and colloids, as well as non-thermal systems such as flowing granular materials and foams. We therefore ask to what extent the concept of effective temperature is valid. Here we investigate this question experimentally in a simple system consisting of a sphere placed on a fine screen in an upward flow of gas; the sphere rolls because of the turbulence it generates in the gas stream. In contrast to many-particle systems, in which it is difficult to measure and predict fluctuations, our system has no particle particle interactions and its dynamics can be captured fully by video imaging. Surprisingly, we find that the sphere behaves exactly like a harmonically bound brownian particle. The random driving force and frequency-dependent drag satisfy the fluctuation-dissipation relation, a cornerstone of statistical mechanics. The statistical mechanics of near-equilibrium systems is therefore unexpectedly useful for studying at least some classes of systems that are driven far from equilibrium. PMID- 14765190 TI - A route to high surface area, porosity and inclusion of large molecules in crystals. AB - One of the outstanding challenges in the field of porous materials is the design and synthesis of chemical structures with exceptionally high surface areas. Such materials are of critical importance to many applications involving catalysis, separation and gas storage. The claim for the highest surface area of a disordered structure is for carbon, at 2,030 m2 g(-1) (ref. 2). Until recently, the largest surface area of an ordered structure was that of zeolite Y, recorded at 904 m2 g(-1) (ref. 3). But with the introduction of metal-organic framework materials, this has been exceeded, with values up to 3,000 m2 g(-1) (refs 4-7). Despite this, no method of determining the upper limit in surface area for a material has yet been found. Here we present a general strategy that has allowed us to realize a structure having by far the highest surface area reported to date. We report the design, synthesis and properties of crystalline Zn4O(1,3,5 benzenetribenzoate)2, a new metal-organic framework with a surface area estimated at 4,500 m2 g(-1). This framework, which we name MOF-177, combines this exceptional level of surface area with an ordered structure that has extra-large pores capable of binding polycyclic organic guest molecules--attributes not previously combined in one material. PMID- 14765191 TI - Hydrogenation and cleavage of dinitrogen to ammonia with a zirconium complex. AB - Molecular nitrogen is relatively inert owing to the strength of its triple bond, nonpolarity and high ionization potential. As a result, the fixation of atmospheric nitrogen to ammonia under mild conditions has remained a challenge to chemists for more than a century. Although the Haber-Bosch process produces over 100 million tons of ammonia annually for the chemical industry and agriculture, it requires high temperature and pressure, in addition to a catalyst, to induce the combination of hydrogen (H2) and nitrogen (N2). Coordination of molecular nitrogen to transition metal complexes can activate and even rupture the strong N N bond under mild conditions, with protonation yielding ammonia in stoichiometric and even catalytic yields. But the assembly of N-H bonds directly from H2 and N2 remains challenging: adding H2 to a metal-N2 complex results in the formation of N2 and metal-hydrogen bonds or, in the case of one zirconium complex, in formation of one N-H bond and a bridging hydride. Here we extend our work on zirconium complexes containing cyclopentadienyl ligands and show that adjustment of the ligands allows direct observation of N-H bond formation from N2 and H2. Subsequent warming of the complex cleaves the N-N bond at 45 degrees C, and continued hydrogenation at 85 degrees C results in complete fixation to ammonia. PMID- 14765192 TI - Low-velocity zone atop the 410-km seismic discontinuity in the northwestern United States. AB - The seismic discontinuity at 410 km depth in the Earth's mantle is generally attributed to the phase transition of (Mg,Fe)2SiO4 (refs 1, 2) from the olivine to wadsleyite structure. Variation in the depth of this discontinuity is often taken as a proxy for mantle temperature owing to its response to thermal perturbations. For example, a cold anomaly would elevate the 410-km discontinuity, because of its positive Clapeyron slope, whereas a warm anomaly would depress the discontinuity. But trade-offs between seismic wave-speed heterogeneity and discontinuity topography often inhibit detailed analysis of these discontinuities, and structure often appears very complicated. Here we simultaneously model seismic refracted waves and scattered waves from the 410-km discontinuity in the western United States to constrain structure in the region. We find a low-velocity zone, with a shear-wave velocity drop of 5%, on top of the 410-km discontinuity beneath the northwestern United States, extending from southwestern Oregon to the northern Basin and Range province. This low-velocity zone has a thickness that varies from 20 to 90 km with rapid lateral variations. Its spatial extent coincides with both an anomalous composition of overlying volcanism and seismic 'receiver-function' observations observed above the region. We interpret the low-velocity zone as a compositional anomaly, possibly due to a dense partial-melt layer, which may be linked to prior subduction of the Farallon plate and back-arc extension. The existence of such a layer could be indicative of high water content in the Earth's transition zone. PMID- 14765193 TI - Mangroves enhance the biomass of coral reef fish communities in the Caribbean. AB - Mangrove forests are one of the world's most threatened tropical ecosystems with global loss exceeding 35% (ref. 1). Juvenile coral reef fish often inhabit mangroves, but the importance of these nurseries to reef fish population dynamics has not been quantified. Indeed, mangroves might be expected to have negligible influence on reef fish communities: juvenile fish can inhabit alternative habitats and fish populations may be regulated by other limiting factors such as larval supply or fishing. Here we show that mangroves are unexpectedly important, serving as an intermediate nursery habitat that may increase the survivorship of young fish. Mangroves in the Caribbean strongly influence the community structure of fish on neighbouring coral reefs. In addition, the biomass of several commercially important species is more than doubled when adult habitat is connected to mangroves. The largest herbivorous fish in the Atlantic, Scarus guacamaia, has a functional dependency on mangroves and has suffered local extinction after mangrove removal. Current rates of mangrove deforestation are likely to have severe deleterious consequences for the ecosystem function, fisheries productivity and resilience of reefs. Conservation efforts should protect connected corridors of mangroves, seagrass beds and coral reefs. PMID- 14765194 TI - Mutations in VKORC1 cause warfarin resistance and multiple coagulation factor deficiency type 2. AB - Coumarin derivatives such as warfarin represent the therapy of choice for the long-term treatment and prevention of thromboembolic events. Coumarins target blood coagulation by inhibiting the vitamin K epoxide reductase multiprotein complex (VKOR). This complex recycles vitamin K 2,3-epoxide to vitamin K hydroquinone, a cofactor that is essential for the post-translational gamma carboxylation of several blood coagulation factors. Despite extensive efforts, the components of the VKOR complex have not been identified. The complex has been proposed to be involved in two heritable human diseases: combined deficiency of vitamin-K-dependent clotting factors type 2 (VKCFD2; Online Mendelian Inheritance in Man (OMIM) 607473), and resistance to coumarin-type anticoagulant drugs (warfarin resistance, WR; OMIM 122700). Here we identify, by using linkage information from three species, the gene vitamin K epoxide reductase complex subunit 1 (VKORC1), which encodes a small transmembrane protein of the endoplasmic reticulum. VKORC1 contains missense mutations in both human disorders and in a warfarin-resistant rat strain. Overexpression of wild-type VKORC1, but not VKORC1 carrying the VKCFD2 mutation, leads to a marked increase in VKOR activity, which is sensitive to warfarin inhibition. PMID- 14765195 TI - Identification of the gene for vitamin K epoxide reductase. AB - Vitamin K epoxide reductase (VKOR) is the target of warfarin, the most widely prescribed anticoagulant for thromboembolic disorders. Although estimated to prevent twenty strokes per induced bleeding episode, warfarin is under-used because of the difficulty of controlling dosage and the fear of inducing bleeding. Although identified in 1974 (ref. 2), the enzyme has yet to be purified or its gene identified. A positional cloning approach has become possible after the mapping of warfarin resistance to rat chromosome 1 (ref. 3) and of vitamin K dependent protein deficiencies to the syntenic region of human chromosome 16 (ref. 4). Localization of VKOR to 190 genes within human chromosome 16p12-q21 narrowed the search to 13 genes encoding candidate transmembrane proteins, and we used short interfering RNA (siRNA) pools against individual genes to test their ability to inhibit VKOR activity in human cells. Here, we report the identification of the gene for VKOR based on specific inhibition of VKOR activity by a single siRNA pool. We confirmed that MGC11276 messenger RNA encodes VKOR through its expression in insect cells and sensitivity to warfarin. The expressed enzyme is 163 amino acids long, with at least one transmembrane domain. Identification of the VKOR gene extends our understanding of blood clotting, and should facilitate development of new anticoagulant drugs. PMID- 14765196 TI - Multiple transport modes of the cardiac Na+/Ca2+ exchanger. AB - The cardiac Na+/Ca2+ exchanger (NCX1; ref. 2) is a bi-directional Ca2+ transporter that contributes to the electrical activity of the heart. When, and if, Ca2+ is exported or imported depends on the Na+/Ca2+ exchange ratio. Whereas a ratio of 3:1 (Na+:Ca2+) has been indicated by Ca2+ flux equilibrium studies, a ratio closer to 4:1 has been indicated by exchange current reversal potentials. Here we show, using an ion-selective electrode technique to quantify ion fluxes in giant patches, that ion flux ratios are approximately 3.2 for maximal transport in either direction. With Na+ and Ca2+ on both sides of the membrane, net current and Ca2+ flux can reverse at different membrane potentials, and inward current can be generated in the absence of cytoplasmic Ca2+, but not Na+. We propose that NCX1 can transport not only 1 Ca2+ or 3 Na+ ions, but also 1 Ca2+ with 1 Na+ ion at a low rate. Therefore, in addition to the major 3:1 transport mode, import of 1 Na+ with 1 Ca2+ defines a Na+-conducting mode that exports 1 Ca2+, and an electroneutral Ca2+ influx mode that exports 3 Na+. The two minor transport modes can potentially determine resting free Ca2+ and background inward current in heart. PMID- 14765197 TI - A proton pore in a potassium channel voltage sensor reveals a focused electric field. AB - Voltage-dependent potassium channels are essential for the generation of nerve impulses. Voltage sensitivity is conferred by charged residues located mainly in the fourth transmembrane segment (S4) of each of the four identical subunits that make up the channel. These charged segments relocate when the potential difference across the membrane changes, controlling the ability of the pore to conduct ions. In the crystal structure of the Aeropyrum pernix potassium channel KvAP, the S4 and part of the third (S3B) transmembrane alpha-helices are connected by a hairpin turn in an arrangement termed the 'voltage-sensor paddle'. This structure was proposed to move through the lipid bilayer during channel activation, transporting positive charges across a large fraction of the membrane. Here we show that replacing the first S4 arginine by histidine in the Shaker potassium channel creates a proton pore when the cell is hyperpolarized. Formation of this pore does not support the paddle model, as protons would not have access to a lipid-buried histidine. We conclude that, at hyperpolarized potentials, water and protons from the internal and external solutions must be separated by a narrow barrier in the channel protein that focuses the electric field to a small voltage-sensitive region. PMID- 14765198 TI - Dephosphorylated SRp38 acts as a splicing repressor in response to heat shock. AB - The cellular response to stresses such as heat shock involves changes in gene expression. It is well known that the splicing of messenger RNA precursors is generally repressed on heat shock, but the factors responsible have not been identified. SRp38 is an SR protein splicing factor that functions as a general repressor of splicing. It is activated by dephosphorylation and required for splicing repression in M-phase cells. Here we show that SRp38 is also dephosphorylated on heat shock and that this dephosphorylation correlates with splicing inhibition. Notably, depletion of SRp38 from heat-shocked cell extracts derepresses splicing, and adding back dephosphorylated SRp38 specifically restores inhibition. We further show that dephosphorylated SRp38 interacts with a U1 small nuclear ribonucleoprotein particle (snRNP) protein, and that this interaction interferes with 5'-splice-site recognition by the U1 snRNP. Finally, SRp38-deficient DT40 cells show an altered cell-cycle profile consistent with a mitotic defect; they are also temperature sensitive and defective in recovery after heat shock. SRp38 thus plays a crucial role in cell survival under stress conditions by inhibiting the splicing machinery. PMID- 14765199 TI - Molecular engineering of a backwards-moving myosin motor. AB - All members of the diverse myosin superfamily have a highly conserved globular motor domain that contains the actin- and nucleotide-binding sites and produces force and movement. The light-chain-binding domain connects the motor domain to a variety of functionally specialized tail domains and amplifies small structural changes in the motor domain through rotation of a lever arm. Myosins move on polarized actin filaments either forwards to the barbed (+) or backwards to the pointed (-) end. Here, we describe the engineering of an artificial backwards moving myosin from three pre-existing molecular building blocks. These blocks are: a forward-moving class I myosin motor domain, a directional inverter formed by a four-helix bundle segment of human guanylate-binding protein-1 and an artificial lever arm formed by two alpha-actinin repeats. Our results prove that reverse-direction movement of myosins can be achieved simply by rotating the direction of the lever arm 180 degrees. PMID- 14765200 TI - Crystal structure and mechanism of a bacterial fluorinating enzyme. AB - Fluorine is the thirteenth most abundant element in the earth's crust, but fluoride concentrations in surface water are low and fluorinated metabolites are extremely rare. The fluoride ion is a potent nucleophile in its desolvated state, but is tightly hydrated in water and effectively inert. Low availability and a lack of chemical reactivity have largely excluded fluoride from biochemistry: in particular, fluorine's high redox potential precludes the haloperoxidase-type mechanism used in the metabolic incorporation of chloride and bromide ions. But fluorinated chemicals are growing in industrial importance, with applications in pharmaceuticals, agrochemicals and materials products. Reactive fluorination reagents requiring specialist process technologies are needed in industry and, although biological catalysts for these processes are highly sought after, only one enzyme that can convert fluoride to organic fluorine has been described. Streptomyces cattleya can form carbon-fluorine bonds and must therefore have evolved an enzyme able to overcome the chemical challenges of using aqueous fluoride. Here we report the sequence and three-dimensional structure of the first native fluorination enzyme, 5'-fluoro-5'-deoxyadenosine synthase, from this organism. Both substrate and products have been observed bound to the enzyme, enabling us to propose a nucleophilic substitution mechanism for this biological fluorination reaction. PMID- 14765201 TI - Cash incentives. PMID- 14765202 TI - All systems go. PMID- 14765203 TI - Graduate journal: conference survival. PMID- 14765204 TI - Nuts and bolts. Selling your skills. PMID- 14765206 TI - Radical prostatectomy for carcinoma of the prostate. AB - Morphologic features of prostatic adenocarcinoma in the radical prostatectomy (RP) specimen are powerful prognostic indicators for prognosis for disease-free survival. This review discusses the methods of sampling of the RP specimen to optimize the detection of these morphologic features, balanced against the added expense of submitting the entire gland for sectioning. Gleason grade, one of the most powerful prognostic factors, is discussed briefly, including the percent pattern 4/5 cancer compared to the standard Gleason grading. Pathologic stage, as defined by the TNM system, is discussed in detail, both in terms of precise histological definition of each category, as well as the associated prognostic implications. Surgical margin status is also important prognostically across all pathologic stages categories. Perineural invasion, which has been used diagnostically in prostate cancer for several decades, has emerged as a very important prognostic indicator as well, as determined by the quantitative aspects of tumor in the perineural space. The effect of tumor volume on prognosis is discussed, as well as the newer concepts of the prognostic significance of zone of origin of the tumor and the presence or absence of intraductal carcinoma. PMID- 14765207 TI - Ns strategies: a highly versatile synthetic method for amines. AB - A highly efficient and versatile synthetic method for amines was established using nitrobenzenesulfonamides (Ns-amides) as both a protecting and activating group. The alkylation of N-monosubstituted Ns-amides either proceeded conventionally or under Mitsunobu conditions to provide the N,N-disubstituted sulfonamides, and the Ns group was removed easily with soft nucleophiles via Meisenheimer complexes to give the corresponding secondary amines. The major advantage of this protocol is that both alkylation and deprotection proceed under mild conditions. Thus, with this methodology, the total synthesis of linear and/or macrocyclic natural polyamines can be accomplished efficiently. PMID- 14765208 TI - Amine elimination synthesis of a titanium(IV) N-heterocyclic carbene complex with short intramolecular Cl...C carbene contacts. AB - The reaction of 1,3-dimesitylimidazolium chloride with Ti(NMe(2))(4) results in the 1,3-dimesitylimadazol-2-ylidene complex of Ti(NMe(2))(2)Cl(2)(3); the X-ray crystal structure of 3 evidences short intramolecular Cl...C(carbene) contacts. PMID- 14765209 TI - Heterogeneous colorimetric sensor for mercuric salts. AB - A novel chemical sensor for the colorimetric detection of mercuric salts is described. The sensor is based on a mesoporous nanocrystalline TiO2 film sensitised with a ruthenium dye; immersion of this film in an aqueous solution of Hg2+ results in a rapid colorimetric response, with both a high selectivity and a sub-micromolar sensitivity. PMID- 14765210 TI - Probing lipase/esterase libraries for lipid A hydrolases--discovery of biocatalysts for the detoxification of bacterially-expressed recombinant protein. AB - In our ongoing efforts to develop new methods for lipopolysaccharide (LPS) detoxification, we have screened lipase/esterase libraries for the ability to deacylate the 2'- and 3'-fatty acid chains from lipid A: the most active esterases were successfully employed to inactivate LPSs in a crude concentrated cell supernatant of E. Coli containing a recombinant single chain antibody (scFv). PMID- 14765211 TI - Novel cofacial oxidative coupling reaction of phosphinine in the presence of CuI and ClO4-. AB - 2,4,6-triphenylphosphinine (TPP) underwent unprecedented cofacial oxidative coupling to form a novel C2-symmetric cage compound, having extremely long C-C bonds. PMID- 14765212 TI - The role of temperature in the synthesis of hybrid inorganic-organic materials: the example of cobalt succinates. AB - Five different cobalt succinate materials synthesized from an identical starting mixture using temperature as the only independent variable show increasing condensation and density at higher synthesis temperatures. PMID- 14765213 TI - H-Bonded complexes of adenine with Rebek imide receptors are stabilised by cation pi interactions and destabilised by stacking with perfluoroaromatics. AB - A series of Rebek imide receptors with naphthalene or heteroaromatic platforms attached by amide or ester linkers have been prepared from the corresponding acyl chloride or anhydride; the X-ray crystal structure of the receptor-derived anhydride reveals a supramolecular H-bonded helix formation in the crystal; the complexes of adenine bound to the receptors by Hoogsteen H-bonding are found to be stabilised by stacking with a methylquinolinium ion, but destabilised by stacking with a perfluorinated naphthalene. PMID- 14765214 TI - Gas-liquid and gas-liquid-solid catalysis in a mesh microreactor. AB - A microstructured mesh contactor that can offer residence time of more than minutes is used for gas-liquid-solid hydrogenations and gas-liquid asymmetric hydrogenations. Applications for catalyst/chiral inductor screening and for kinetic data acquisition are demonstrated. PMID- 14765215 TI - First synthesis of a series of core-modified tetrabenzoporphyrins. AB - Successful synthesis of a series of highly conjugated porphyrin analogues, including thia-, dithia- and oxathia-tetrabenzoporphyrins, and their optical properties are reported. PMID- 14765216 TI - Evidences of the origin of N2O in the high-temperature NH3 oxidation over Pt-Rh gauze. AB - Transient isotopic experiments reveal that the mechanism of N2O formation in the high-temperature NH3 oxidation over Pt-Rh gauze involves the reaction of adsorbed ammonia intermediate species (NHx) and NO. PMID- 14765217 TI - Designed double layer assembly: a three-dimensional open framework with two types of cavities by connection of infinite two-dimensional bilayer. AB - A photoluminescent three-dimensional coordination polymer containing two types of cavities is prepared by the covalent linkage of 2D bilayer motif with linear connecting ligand. PMID- 14765218 TI - A new type of entanglement involving one-dimensional ribbons of rings catenated to a three-dimensional network in the nanoporous structure of [Co(bix)2(H2O)2](SO4).7H2O [bix = 1,4-bis(imidazol-1-ylmethyl)benzene]. AB - Co(II) sulfate reacts with the flexible ligand 1,4-bis(imidazol-1 ylmethyl)benzene (bix) to yield the coordination network [Co(bix)2(H2O)2](SO4).7H2O, containing polymeric ribbons of rings which penetrate and catenate a 3D single frame of the CdSO4 topology, to produce an open-channel entangled architecture with nanoporous behaviour. PMID- 14765219 TI - Lactose-appended schizophyllan is a potential candidate as a hepatocyte-targeted antisense carrier. AB - A schizophyllan (beta-1,3-glucan) derivative carrying lactose-appendages prepared by reductive amination can form stable macromolecular complexes with polynucleotides, shows excellent affinity with a lactose-binding lectin, and effectively mediates gene transfection into hepatocytes. PMID- 14765220 TI - A novel chiral terpyridine macrocycle as a fluorescent sensor for enantioselective recognition of amino acid derivatives. AB - Terpyridine macrocycle 1 is shown to be a strong chelating agent for organic ammonium salts and also a useful chromophore in fluorescent sensing. It exhibits very good enantioselectivity (K(obs)(S)/K(obs)(R)= 3.8) in chiral discrimination of alpha-phenylglycine methyl ester hydrochloride (PhEtOMe). PMID- 14765221 TI - Preparation of alkyl-surface functionalized germanium quantum dots via thermally initiated hydrogermylation. AB - A new, thermally initiated hydrogermylation-based method for the synthesis and surface functionalization of air- and moisture-stable germanium quantum dots is reported. PMID- 14765222 TI - A (bpy)2Ru-coordinated dehydro[12]annulene with exotopically fused diimine binding sites. AB - Synthesis and electronic properties of a dinuclear (bpy)(2)Ru(II) polypyridyl complex are described in which the bridging ligand consists of two dipyridophenazines fused to a formally antiaromatic dehydro[12]annulene and where the electronic and electrochemical properties of the complex are markedly influenced by the cyclic all-carbon core. PMID- 14765223 TI - Parallel synthesis and characterization of photoelectrochemically and electrochromically active tungsten-molybdenum oxides. AB - Single phase tungsten-molybdenum mixed oxide films (W(1-x)Mo(x)O(3)) were successfully synthesized by automated parallel electrodeposition, and distinct structural changes were observed as a function of composition. A monoclinic structure (beta-phase) was observed in mixed oxides with less than 90% Mo, and above 90% Mo, orthorhombic structure (alpha-phase) was identified. PMID- 14765224 TI - The nature of oxygen exchange in ZrW2O8 revealed by two-dimensional solid-state 17O NMR. AB - 17O magic angle spinning (MAS) NMR has been used to determine the nature of oxygen exchange in ZrW(2)O(8). A highly effective isotopic labelling technique has been developed and 1D NMR and 2D exchange spectroscopy (EXSY) experiments have revealed that mutual exchange occurs between all oxygen sites, even at temperatures considerably below the alpha to beta order-disorder phase transition. PMID- 14765225 TI - Novel 1,3-dipolar cycloaddition of diazocarbonyl compounds to alkynes catalyzed by InCl3 in water. AB - The first intermolecular 1,3-dipolar cycloaddition of diazocarbonyl compounds with alkynes was developed by using an InCl(3) catalyzed cycloaddition in water. The reaction was found to proceed by a domino 1,3-dipolar cycloaddition-hydrogen (alkyl or aryl) migration. PMID- 14765226 TI - Self-organization of a tetrasubstituted tetrathiafulvalene (TTF) in a silica based hybrid organic-inorganic material. AB - A hybrid organic inorganic nanostructured material containing a TTF core substituted by four arms exhibited a high level of both condensation at silicon (96%) and self-organization as evidenced by X-ray diffraction and an unprecedented birefringent behaviour. PMID- 14765227 TI - Synthesis and catalytic activity of a poly(N,N-dialkylcarbodiimide)/palladium nanoparticle composite: a case in the Suzuki coupling reaction using microwave and conventional heating. AB - Poly(N,N-dialkylcarbodiimide) was found to be an effective polymeric ligand system for preparing and stabilizing palladium nanoparticles (1-5 nm). The composite material prepared in situ was found to be a robust catalyst for the Suzuki coupling reaction under microwave or regular heating. PMID- 14765228 TI - Improved photoelectrochemical performance of electrodeposited ZnO/EosinY hybrid thin films by dye re-adsorption. AB - Dye desorption and re-adsorption post treatments on electrochemically self assembled nanoporous ZnO/eosinY hybrid thin films lead to a large improvement of the dye-sensitized photoelectrochemical performance, achieving an incident photon to current conversion efficiency up to 90%. PMID- 14765229 TI - Thermal reactions of an overcrowded germacyclopropabenzene with group 6 metal hexacarbonyl complexes [M(CO)6](M = Cr, Mo, and W): a novel mode of CO insertion leading to the formation of cyclic germoxycarbene metal complexes. AB - Thermal reactions of an overcrowded germacyclopropabenzene with hexacarbonyl complexes of group 6 metals resulted in the formation of the corresponding cyclic germoxycarbene complexes, i.e., 1,2-oxagermolan-5-ylidene pentacarbonyl metal complexes, via a novel mode of CO insertion reaction toward cyclopropabenzene derivatives. PMID- 14765230 TI - Electrochemical wiring of alpha, omega-alkanedithiol molecules into an electrical circuit. AB - The intrinsic electrical conductivity of alpha, omega-alkanedithiol increases if both ends of the molecule are covalently bonded to metallic contacts. PMID- 14765231 TI - First total synthesis of murisolin. AB - The first and concise total synthesis of murisolin (1) was accomplished using asymmetric alkynylation and Sonogashira coupling as the key steps. The threo/trans/threo-type THF ring moiety was constructed with excellent stereoselectivity by asymmetric alkynylation of 1,6-heptadiyne to alpha tetrahydrofuranic aldehyde, which was also prepared via the asymmetric alkynylation. PMID- 14765232 TI - Construction of porphyrin-cyclodextrin self-assembly with molecular wedge. AB - A new host porphyrin bearing four permethyl-beta-cyclodextrin moieties for multi porphyrin assembly forms a unique 2 : 2 assembly with the tetra-anion of tetrakis(p-sulfonylphenyl)porphyrin (TPPS) in aqueous solution. PMID- 14765233 TI - Facile fabrication of composites of platinum nanoparticles and amorphous carbon films by catalyzed carbonization of cellulose fibers. AB - Composites of platinum nanoparticles and amorphous carbon films have been facilely fabricated by catalyzed carbonization of cellulose fibers. PMID- 14765234 TI - Generation of strong, homochiral bases by electrochemical reduction of phenazine derivatives. AB - Electrochemical reduction of enantiomerically pure amino- and alkoxy-phenazine derivatives forms strongly basic radical anions which give asymmetric induction in the conversion of 3,4-epoxytetrahydrothiophene-1,1-dioxide into the allylic ester with facile regeneration of the phenazine. PMID- 14765235 TI - Zinc metalloporphyrin-functionalised nanoparticle anion sensors. AB - Disulfide-functionalised zinc metalloporphyrins self-assembled on gold nanoparticles exhibit remarkable, surface-enhanced, anion binding affinities as compared to the free metalloporphyrin. PMID- 14765236 TI - Mn3(HCOO)6: a 3D porous magnet of diamond framework with nodes of Mn-centered MnMn4 tetrahedron and guest-modulated ordering temperature. AB - Mn3(HCOO)6, a 3D highly stable and flexible porous diamondoid framework based on Mn-centered MnMn4 tetrahedral nodes, exhibits a wide spectrum of guest inclusion behaviour and long-range magnetic ordering with guest-modulated critical temperature. PMID- 14765237 TI - Temperature-controlled hydrothermal synthesis of a 2D ferromagnetic coordination bilayered polymer and a novel 3D network with inorganic Co3(OH)2 ferrimagnetic chains. AB - A 2D bilayered ferromagnetic coordination polymer, generated by lower-temperature hydrothermal reactions of cobalt(II) salt with the 3,4-pyridinedicarboxylate dianion, was controlled to transform into a 3D magnetic coordination network with Co3(OH)2 ferrimagnetic chains at a higher temperature. PMID- 14765238 TI - Metal ion induced allosteric transition in the catalytic activity of an artificial phosphodiesterase. AB - An artificial phosphodiesterase (1) bearing two kinds of metal binding sites, a catalytic site and a regulatory bipyridine site showed a unique allosteric transition in the catalytic activity against the metal concentration. PMID- 14765239 TI - Cross metathesis functionalization of polyolefins. AB - A cross metathesis strategy is reported for the post-polymerization functionalization of the pendant vinyl groups present in a range of polyolefin architectures. This represents a general strategy for the synthesis of tailored random and block copolymers as well as homopolymers. PMID- 14765240 TI - [VO(H2O)5]H[PMo12O40]-catalyzed nitration of alkanes with nitric acid. AB - [VO(H2O)5]H[PMo12O40], which contains vanadyl counter cations and PMo12O40(3-), can act as a catalyst for the nitration of various alkanes including alkylbenzenes using nitric acid as a nitrating agent in acetic acid at 356 K. PMID- 14765241 TI - A trans-tetrahydrobenzoxanthene receptor for the resolution of racemic mixtures of sulfonylamino acids. AB - An enantioselective cleft-type receptor for sulfonylamino acids has been prepared and its use for the resolution of the amino acid racemic mixture is shown. PMID- 14765242 TI - Redox sensing of anions in pure aqueous environment by ferrocene-containing 4,4' bipyridinium-based receptors and polymer films. AB - Selective voltammetric sensing of ATP(2-) anions in water was achieved using ferrocene-viologen based redox active receptors and related polymer films. PMID- 14765244 TI - Enantioselective Friedel-Crafts reaction of indoles with arylidene malonates catalyzed by iPr-bisoxazoline-Cu(OTf)2. AB - The cheap and simple (i)Pr-bisoxazoline-Cu(OTf)(2) proves to be an efficient catalyst in the asymmetric Friedel-Crafts reaction of indole with arylidene malonates. In (i)BuOH, the S-enantiomer was obtained in up to 97% ee, while the opposite enantiomer was obtained in up to 78% ee in CH(2)Cl(2) or TTCE. PMID- 14765243 TI - Interaction of modified liposomes with Bacillus spores. AB - The interaction between liposomes modified with a particular peptide sequence and Bacillus subtilis spores was experimentally observed as (1) an increase in the average diameter of spore-related particles, and (2) the formation of dense and structured shells around the spores at higher concentrations of liposomes. PMID- 14765245 TI - Discovery and evaluation of highly active imidotitanium ethylene polymerisation catalysts using high throughput catalyst screening. AB - A family of ca. 50 imidotitanium precatalysts [Ti(NR)(Me(3)[9]aneN(3))Cl(2)](R = alkyl or aryl; Me(3)[9]aneN(3)= 1,4,7-trimethyltriazacyclononane) were prepared in good yields using semi-automated procedures; high-throughput screening techniques identified seven highly active ethylene polymerisation precatalysts with activities in the range ca. 3 400 to 10 000 kg(PE) mol(-1) h(-1) bar(-1). PMID- 14765246 TI - Bridging interaction between a water drop stabilised by solid particles and a planar oil/water interface. AB - We demonstrate that the particle mediated interaction between a pendant water drop, covered by a latex particle monolayer, and a planar decane/water interface leads to bridging, analogous to flocculation in solid-stabilised emulsions. The results also provide information about the particle contact angle at the oil/water interface. PMID- 14765247 TI - Chemoselective signalling of selected phospho-anions using lanthanide luminescence. AB - Selectivity in the binding of phosphorylated tyrosine residues to aqua-lanthanide complexes is signalled by changes in spectral form by luminescence emission and 1H NMR spectroscopy. PMID- 14765248 TI - Co2+-exchanged faujasite zeolites as efficient heterogeneous catalysts for epoxidation of styrene with molecular oxygen. AB - Co2+-exchanged faujasite zeolites can efficiently catalyze the epoxidation of styrene with molecular oxygen, and the Co2+ ions located in supercages are suggested to account for the activation of O2 for the epoxidation of styrene. PMID- 14765249 TI - Proton transfer versus redox modulation in thiourea-phenanthrenequinone molecular and polymeric complexes. AB - Phenanthrenequinone undergoes highly efficient proton transfer processes in the presence of a thiourea-functionalised polystyrene copolymer whereas interactions with a similar benzyl-thiourea monomer show strong redox modulation of the quinone without proton transfer. PMID- 14765250 TI - Thermal formation of serine octamer ions. AB - Vigorous evaporation of aqueous serine solutions yields abundant protonated serine octamer ions. So does pyrolysis of L-serine crystals in a corona discharge. PMID- 14765251 TI - Copper- and palladium-catalyzed intramolecular C-S bond formation: a convenient synthesis of 2-aminobenzothiazoles. AB - Copper- and palladium-catalyzed intramolecular C-S bond formation by cross coupling between an aryl halide and thiourea functionality is demonstrated for the synthesis of 2-aminobenzothiazoles, wherein the Cu-catalyzed protocol is generally superior and more cost effective than the Pd-catalyzed protocol; the Cu catalyzed reaction also further expands recent studies exploring the utility of Cu salts as replacements for Pd in carbon-heteroatom bond-forming reactions. A one-pot variant combining the synthesis of the thiourea and the cyclization was also demonstrated. PMID- 14765252 TI - Stabilization of D5h and C2v valence tautomers of the croconate dianion. AB - The non-benzenoid aromatic D(5h) and enediolate C(2v) form of C5O5(2-) have been stabilized by hydrogen bonding with urea and 1,3-dimethylurea, respectively, in the host lattices of two novel crystalline inclusion compounds. PMID- 14765253 TI - Interfacial polymerisation of anilinium at Langmuir monolayers. AB - Anilinium is strongly adsorbed at monolayers of the phospholipid L-alpha dimyristoylphosphatidic acid (DMPA) and hexadecanesulfonic acid (HDSA) at the air water interface, and undergoes chemical polymerisation under conditions where bulk polymerisation does not occur. PMID- 14765254 TI - The biosynthesis of crucifer phytoalexins: unprecedented incorporation of a 1 methoxyindolyl precursor. AB - The first biosynthetic studies revealing that 1-methoxy-3-indolylacetaldehyde oxime is an early precursor of 1-methoxyindole containing phytoalexins. PMID- 14765255 TI - The parallel synthesis of a disaccharide library using a solid phase, peptide templated strategy. AB - A novel strategy for the synthesis of oligosaccharides, involving the use of a solid phase peptide template, has been successfully applied to the construction of a twelve member disaccharide library. PMID- 14765256 TI - An organically templated Co(II) sulfate with the kagome lattice. AB - An amine-templated cobalt(II) sulfate with the kagome lattice, prepared for the first time, exhibits magnetic properties comparable to those of the analogous Fe(III) compounds. PMID- 14765257 TI - Synthesis and properties of a new class of nitrogen-rich multinuclear[m.n] ferrocenophanes. AB - The synthesis of the first tetraaza[3,3]ferrocenophane, from the previously unreported 1,1'-bis(triphenylphosphoranylidenamino)ferrocene and its conversion into a nitrogen-rich [2,2]bis[3,3]ferrocenophane is described. The electronic behaviour of these multinuclear ferrocenophanes indicates a strong electronic coupling between the iron centers. PMID- 14765258 TI - Structures and reactivity of synthetic zinc(II) complexes resembling the active sites and reaction intermediates of aminopeptidases. AB - Herein we report the first crystallographic characterization and hydrolysis of a synthetic zinc(II) complex that resembles the active site and reaction intermediates proposed for aminopeptidases. PMID- 14765259 TI - Surface step structure of Ag13OsO6, experimental evidence for Ag13 cluster building blocks. AB - The surface of single crystal Ag(13)OsO(6) has been investigated using atomic force microscopy. Growth spirals with very large flat terraces, separated by steps of equal height, have been observed. The measured step height of approximately 6.7 Angstrom corresponds to the diameter of one Ag13-icosahedron and identifies the cluster as the key structural building block. PMID- 14765260 TI - New insights on the asymmetric hydroboration of perfluoroalkenes. AB - Enantioselective access to Markovnikov regioisomeric perfluoroalcohols is achieved in the presence of chiral cationic rhodium complexes and specific hydroborating reagents. PMID- 14765261 TI - A novel synthesis of chiral rotaxanes via covalent bond formation. AB - Chiral rotaxanes composed of the asymmetric crownophane incorporating two hydroxy groups as a rotor moiety and the asymmetric axis were effectively synthesized via covalent bond formation, i.e. tandem Claisen rearrangement, esterification, and aminolysis. PMID- 14765262 TI - Remarkable thermodynamic stability toward hydrolysis of tripodal titanium alkoxides. AB - The monomeric titanium(IV) hydroxide complex, LTi(OH)(LH(3)= tris(2-hydroxy-3,5 di-tert-butylbenzyl)amine), which is sterically inhibited from condensation to a mu-oxo dimer, cannot be prepared by hydrolysis of the alkoxide, with K(eq)= 0.012 for hydrolysis of the titanium methoxide in THF. PMID- 14765263 TI - A mild and efficient procedure for alpha-bromination of ketones using N bromosuccinimide catalysed by ammonium acetate. AB - Cyclic ketones reacted with N-bromosuccinimide (NBS) catalysed by NH(4)OAc in Et(2)O at 25 degrees C to give the corresponding alpha-brominated ketones in good yields, while acyclic ketones were efficiently brominated in CCl(4) at 80 degrees C. PMID- 14765264 TI - Synthesis of methanesulfonyl chloride (MSC) from methane and sulfuryl chloride. AB - Methane is transformed selectively to methanesulfonyl chloride at low temperature by liquid-phase reaction of methane with SO(2)Cl(2) in the presence of a free radical initiator and a promoter using 100% H(2)SO(4) as the solvent. PMID- 14765265 TI - A copper-complexed rotaxane in motion: pirouetting of the ring on the millisecond timescale. AB - A new bistable rotaxane, consisting of a 2,2'-bipyridine-containing thread and a ring incorporating both a bidentate chelate and a tridentate fragment, has been prepared; this complex undergoes an electrochemically driven pirouetting motion of the ring around the axis which takes place on the millisecond timescale, i.e. several orders of magnitude faster than the other copper-based machines previously described. PMID- 14765266 TI - "Stromatogenesis" and tumor progression. AB - "Stromatogenesis" is the formation of new stroma occurring, in parallel with the neoplastic process, at sites of active tumor invasion, i.e., at the free surface of a developing exophytic tumor, at the invading tumor front of an advancing endophytic tumor, and at sites of tumor metastasis, wherein the newly formed stroma disrupts the continuity of normal structures, cleaving paths for the invading tumor cells. Stroma is also present at the heart of the tumor, but only as a secondary event following tumor advancement and subsequent incorporation of its periphery into inner tumor areas. The new stroma, composed of stromal cells and extracellular matrix (ECM), is loose and edematous at the expanding tumor fronts, and rather dense in central tumor areas and sites of tumor metastasis. The stromal cells facing tumor invasion are intensely proliferating (high MIB-1 index) spindle-shaped cells, alpha-smooth muscle actin positive, and loaded with thymidine phosphorylase (TP) and SPARC (secreted protein acidic and rich in cysteine). The associated ECM is rich in collagen III, SPARC, and new blood vessels (CD31) but is depleted of collagen I and fibronectin. These constitutional changes render stromatogenesis amenable to tumor cell invasion and are, in cases of incipient neoplasia, a prospective criterion of early stromal invasion. Other stromal cell or ECM constituents, such as the lactate dehydrogenase-5 (LDH-5), the acidic fibroblast growth factor (aFGF), the basic FGF (bFGF), and the collagens II and IV, remain unchanged, and others are negative: myosin, desmin, S-100 protein and epidermal growth factor receptor (EGFR). The mechanism of stromatogenesis is obscure but is probably stimulated by specific stromatogenic growth factors, released by neoplastic and inflammatory cells. It appears that the process is neither neoplastic nor reactive, but rather is a, hereto unexplained, phenomenon of host's complicity in tumor progression. PMID- 14765267 TI - Tenascin C and cathepsin d expression in adipocytic tumors: an immunohistochemical investigation of 43 cases. AB - Cathepsin D (CatD) and tenascin C (Tn-C) have been implicated in invasion and metastasis of carcinomas. However, little is known about CatD and Tn-C distribution in mesenchymal tumors. Therefore, we aimed to investigate the expression of Tn-C and CatD in adipocytic tumors. Tn-C and CatD expressions in 27 lipomas, 5 atypical lipomatous tumor/well-differentiated liposarcomas (ALT/WDLS) and 11 liposarcomas (LS) were evaluated by immunohistochemistry and scored semiquantitatively. CatD expression was higher in ALT/WDLS than in lipomas (p<0.001), and higher in LS than in ALT/WDLS (p=0.009). Stromal Tn-C expression was higher in ALT/WDLS than in lipomas (p=0.02), but no difference was observed between ALT/WDLS and LS (p>0.05). These results indicate that Tn-C and CatD may be markers of locally aggressive or malignant behavior in adipocytic tumors. PMID- 14765268 TI - Desmoplastic melanoma of the head and neck: histopathologic and immunohistochemical study of 28 cases. AB - Studies of the immunohistochemical profiles and clinical course of desmoplastic melanoma have produced conflicting results. We identified 28 cases of desmoplastic melanoma after a search of our files for spindle cell neoplasms of the head and neck from 1960 through 1995. The 17 male (61%) and 11 female (39%) patients averaged 65 years of age. The cheek was the most common location (12 cases, 43%). The average length of follow-up was 5 years. Overall 5-year survival rate was 46%. Melan A and tyrosinase positivity (P = 0.0195), smooth muscle actin positivity (P = 0.0328), tumor size (P = 0.0297), and tumor thickness (P = 0.0419) were significantly associated with local progression-free survival. No histologic or immunohistochemical marker was associated with overall or metastasis-free survival. PMID- 14765269 TI - Clinical implication of idiopathic plasmacytic lymphadenopathy with polyclonal hypergammaglobulinemia: a report of 16 cases. AB - Idiopathic plasmacytic lymphadenopathy (IPL) with polyclonal hyperimmunoglobulinemia is considered identical to multicentric Castleman's disease (MCD) reported in western countries. Clinically, both IPL and MCD are characterized by multicentric lymphadenopathy, prominent polyclonal hypergammaglobulinemia, elevated erythrocyte sedimentation rate, elevated serum interleukin-6 concentration, bone marrow plasmacytosis, and various abnormal laboratory data such as anemia and positive autoantibodies. However, IPL has a significantly better 5-year survival rate than that of MCD. Moreover, none of the present 16 cases developed Kaposi's sarcoma or B-cell lymphoma. Histologically, the interfollicular area contains a sheet of polytypic mature plasma cells in both IPL and MCD. In MCD, the majority of lymphoid follicles had hyaline-vascular germinal centers. However, lymphoid follicles of IPL usually exhibit a hyperplastic germinal center. Immunostaining also demonstrated a normal/reactive follicular dendritic cell network pattern in the germinal center of IPL. Moreover, there were no human herpes virus-8-positive cells detected by immunohistochemistry. The overall clinicopathologic and immunohistochemical findings of our 16 cases suggest that IPL is distinct from MCD reported in Western countries. PMID- 14765270 TI - T-cell-rich B-cell lymphoma presenting in the spleen: a clinicopathologic analysis of 3 cases. AB - We review the clinical, pathologic, and molecular genetic features of 3 splenic T cell-rich B-cell lymphomas and discuss their differential diagnosis. All patients presented with symptomatic splenomegaly and underwent diagnostic/therapeutic splenectomy. Microscopically, the spleen in all cases showed a micronodular proliferation of lymphoid cells. A proportion of the nodules demonstrated central hyalinization or sclerosis. There was also an exuberant extramedullary hematopoiesis. On immunohistochemical stain, the nodules consisted predominantly of small T cells with scattered large atypical B cells. The clonal nature of the atypical B cells was confirmed by polymerase chain reaction assays for immunoglobulin heavy-chain gene rearrangement. In the H&E sections, the differential diagnoses included Hodgkin's lymphoma, follicular lymphoma, peripheral T-cell lymphoma, and nonneoplastic granulomatous process. The presence of exuberant extramedullary hematopoiesis also raised the possibility of a chronic myeloproliferative disorder. The combined morphologic, immunohistochemical, and molecular genetic data are essential for a correct diagnosis of splenic T-cell-rich B-cell lymphoma. PMID- 14765271 TI - Primary cutaneous leiomyosarcoma: a clinicopathological and immunohistochemical study of 7 cases. AB - Primary cutaneous leiomyosarcomas are rare tumors, few series being reported in the current literature. A retrospective study of 7 cases was undertaken to understand the clinicopathological characteristics of these neoplasms and some of their molecular mutations. Histologically, a well-differentiated proliferation of cells of smooth muscle derivation was evident in all cases. The number of mitoses was considered the most important criterion of malignancy (more than 2 for 10 HPF). Smooth muscle actin, desmin, and vimentin were positive in all cases. Immunohistochemical analysis also revealed a positivity for p53 in 3 cases and no reaction for retinoblastoma protein. Research for Epstein-Barr virus was negative in all cases. Three patients developed local recurrences owing to incomplete surgical excision. Recurrent tumors were more atypical and located deeper. No distant metastases were observed. Our results emphasize that cutaneous leiomyosarcomas have an indolent biological course if treated by surgical excision with wide margins. Molecular abnormalities involving tumor suppressor genes are probably involved. PMID- 14765272 TI - Microscopic colitis with giant cells: a rare new histopathologic subtype? AB - Collagenous and lymphocytic colitis might be part of the same disease spectrum. In this report, we present a histopathologic subtype of microscopic colitis characterized by the presence of subepithelial multinucleated giant cells. This reaction is very unusual and not explicable by any underlying disease process or previous treatment. Among 490 cases of microscopic colitis (MC) diagnosed between 1992 and 2002, we found 2 cases with macrophages and giant cells (0.4%). One case of lymphocytic colitis (LC) and 1 case of collagenous colitis (CC) presented aggregates of macrophages and giant cells located in the superficial part of the lamina propria. Infectious or non-infectious colonic granulomatous diseases were excluded on histologic, clinical, and biological grounds. The recognition of this feature in an MC is important to avoid the diagnosis of granulomatous infectious or idiopathic colitis such as Crohn's disease. Even if very unusual, this subtype of MC evolves favorably since the 2 patients responded well to corticosteroid treatment. PMID- 14765275 TI - Myoid angioendothelioma of the spleen. AB - Although vascular neoplasms of the spleen are rare, they are the most common nonhemopoietic proliferation of the organ, and include hemangiomas, lymphangiomas, hamartomas, littoral cell angiomas, hemangioendotheliomas, and angiosarcomas, as well as the recently described myoid angioendothelioma (MA). MA is an uncommon, benign tumor of the spleen, which is morphologically characterized by a composite of vascular spaces and stromal cells with myoid features. In 1999, in the only report of this unusual neoplasm, Kraus and Dehner described the features of 3 cases. We present another case of MA of the spleen occurring in a 51-year-old man that demonstrated the characteristic morphologic and immunohistochemical features of this neoplasm. In addition to the features described by Kraus and Dehner, our case also displayed the previously unreported findings of focal spindling of the stromal cells and scattered S100-positive cells in the stroma. The case was further unique in having a central stellate scar. Careful attention to histology, possibly with the aid of immunohistochemistry, should distinguish other splenic neoplasms from MA. Although MA is a morphologically distinct lesion, its histologic spectrum, biological behavior, and relationship to other vascular tumors are yet to be fully discovered. It is hoped that the recognition of further cases, and the use of newer molecular technologies, will help better define the nosological position and implications of the diagnosis of this unusual tumor. PMID- 14765276 TI - Adrenal cortical adenoma with excess black pigment deposition, combined with myelolipoma and clinical Cushing's syndrome. AB - We report a case of a functional adenoma with excess black pigment deposition and myelolipoma in the same adrenal gland in a 58-year-old woman. The patient presented with gastrointestinal bleeding, and after being diagnosed with colonic diverticulosis, underwent a total colectomy. An abdominal computerized tomographic (CT) scan during her work-up showed a right adrenal mass consistent with myelolipoma. Postoperatively, the patient was diagnosed with Cushing's syndrome and underwent a right adrenalectomy. The adrenalectomy specimen consisted of a dark brown and golden-yellow adrenal adenoma, myelolipoma, and atrophic adrenal gland. Immunostains indicated that the dark brown adenoma component was responsible for the patient's hypercortisolism. Co-occurrence of a functional black adenoma and a well-developed myelolipoma has not been reported in the literature. We describe the significant findings of this case, together with a review of the literature. PMID- 14765277 TI - Adenocarcinoma and dysplasia in an ileal neobladder after ileocystoplasty for interstitial cystitis. AB - We report a 54-year-old woman with an adenocarcinoma of an ileal neobladder arising upon a background of ileal mucosal dysplasia. We believe that no case study or report has previously documented neobladder ileal mucosal dysplasia adjacent to an ileal neobladder adenocarcinoma. This observation supports the current hypothesis that ileal neobladders are dynamic environments for potential malignancy, and moreover, suggests a sequence of morphologic and molecular derangements similar to that seen in colorectal carcinoma. Those patients status post ileal neobladder are at risk for glandular dysplasia and malignancy and should be followed closely. PMID- 14765278 TI - Malignant fibrous histiocytoma arising within a bone infarct in a patient with sickle cell trait. AB - Sarcoma associated with osteonecrosis or bone infarction is a rare but well documented pathological event. In this report, a 69-year-old man with sickle cell trait presented with malignant fibrous histiocytoma (MFH) in his distal tibia. The resected tumor was found in association with a large medullary infarct that extended 10 cm proximal from the tumor site. Bone infarcts can be caused by a number of processes including corticosteroid overuse, alcoholism, dysbarism, and hemoglobinopathies such as sickle cell disease. Patients with sickle cell anemia often develop osteonecrosis, but osteonecrosis has also been reported in people with sickle cell trait, albeit much more rarely. Our patient is only the third reported case of infarct-related bone sarcoma in a patient with sickle cell trait. Bone infarction may be a rare though serious consequence of sickle cell trait. PMID- 14765279 TI - Periprostatic subendothelial intravascular granulomatosis: a mimic of high-grade intravascular prostatic adenocarcinoma. AB - A rare case of intravascular granulomatous inflammation mimicking intravascular prostatic adenocarcinoma is reported. To the author's knowledge, there have been no previous reports of prostatic or periprostatic intravascular granulomatous inflammation. A 67-year-old man presented with elevated serum prostate specific antigen (PSA) and was found to have a high-grade adenocarcinoma of the prostate. A radical prostatectomy revealed intravascular subendothelial granulomatous inflammation mimicking vascular invasion of a high-grade adenocarcinoma found elsewhere in the prostate. Immunoperoxidase stains confirmed that the subendothelial infiltrate was composed of histiocytes and not tumor cells. Periprostatic subendothelial intravascular granulomatosis is a rare event, which may mimic vascular involvement of high-grade prostatic adenocarcinoma and may result from a previous needle biopsy of the prostate. Possible mechanisms for this finding are discussed. It is important to distinguish this process from high grade prostatic adenocarcinoma involving blood vessels for obvious clinical reasons. PMID- 14765280 TI - Focal divergent chondrosarcomatous differentiation in a primary pleomorphic liposarcoma and expression of transforming growth factor beta. AB - A rare case of primary pleomorphic liposarcoma of the thigh with a myxoid component, in which divergent differentiation to a well-differentiated chondrosarcoma was focally present, is described. Presence of heterologous elements has mainly been recognized in the context of dedifferentiated liposarcomas. Few cases of benign mesenchymal tissue have also been reported in well-differentiated and myxoid liposarcomas, while divergent sarcomatous differentiation in liposarcomas appears to be also rare in the absence of dedifferentiation. Positive immunostaining of transforming growth factor-beta, which seems to play a role in the formation of bone and cartilage, was demonstrated in our case. Review of the existing literature on the subject has been carried out. PMID- 14765281 TI - Multicystic mesothelioma of the liver with secondary involvement of peritoneum and inguinal region. AB - A case of multicystic mesothelioma of the liver with secondary involvement of the pelvic peritoneum and the inguinal region is presented. The case is of interest because of its unusual location and peculiar biological behavior. PMID- 14765284 TI - Evaluation of the effects of East Indian sandalwood oil and alpha-santalol on humans after transdermal absorption. AB - The aim of the study was to investigate the effects of East Indian sandalwood oil ( Santalum album, Santalaceae) and alpha-santalol on physiological parameters as well as on mental and emotional conditions in healthy human subjects after transdermal absorption. In order to exclude any olfactory stimulation, the inhalation of the fragrances was prevented by breathing masks. Eight physiological parameters, i. e., blood oxygen saturation, blood pressure, breathing rate, eye-blink rate, pulse rate, skin conductance, skin temperature, and surface electromyogram were recorded. Subjective mental and emotional condition was assessed by means of rating scales. While alpha-santalol caused significant physiological changes which are interpreted in terms of a relaxing/sedative effect, sandalwood oil provoked physiological deactivation but behavioral activation. These findings are likely to represent an uncoupling of physiological and behavioral arousal processes by sandalwood oil. PMID- 14765282 TI - A portable array biosensor for detecting multiple analytes in complex samples. AB - The Multi-Analyte Array Biosensor (MAAB) has been developed at the Naval Research Laboratory (NRL) with the goal of simultaneously detecting and identifying multiple target agents in complex samples with minimal user manipulation. This paper will focus on recent improvements in the biochemical and engineering aspects of this instrument. These improvements have enabled the expansion of the repertoire of analytes detected to include Salmonella typhimurium and Listeria monocytogenes, and also expanded the different sample matrices tested. Furthermore, all components of the biochemical assays could be prepared well in advance of sample testing, resulting in a "plug-and-play" methodology. Simultaneous detection of three toxins (ricin, staphylococcal enterotoxin B, and cholera toxin) was demonstrated using a novel fluidics cube module that limits the number of manipulations to only the initial sample loading. This work demonstrates the utility of the MAAB for rapid analysis of complex samples with multianalyte capability, with a minimum of operator manipulations required for either sample preparation or final analysis. PMID- 14765285 TI - Cancer chemopreventive activity of rotenoids from Derris trifoliata. AB - A study of the chemical constituents of the stems of Derris trifoliata Lour. (Leguminosae) led to the isolation and identification of one new rotenoid, 6aalpha,12aalpha-12a-hydroxyelliptone ( 3), together with five other known rotenoids. In a search for novel cancer chemopreventive agents (anti-tumor promoters), we carried out a primary screening of five of the rotenoids isolated from the plant for their inhibitory effects on Epstein-Barr virus early antigen (EBV-EA) activation induced by 12- O-tetradecanoylphorbol 13-acetate (TPA) in Raji cells. The inhibitory activity of 3 was found to be equivalent to that of beta-carotene without any cytotoxicity. Deguelin ( 4) and alpha-toxicarol ( 5) exhibited a marked inhibitory effect on mouse skin tumor promotion in an in vivo two-stage carcinogenesis test. This investigation indicated that rotenoids might be valuable anti-tumor promoters. PMID- 14765286 TI - Rhein inhibits the growth and induces the apoptosis of Hep G2 cells. AB - The effects of rhein on the human hepatoblastoma G2 (Hep G2) cell line were investigated in this study. The results showed that rhein not only inhibited Hep G2 cell growth but also induced apoptosis and blocked cell cycle progression in the G1 phase. An ELISA assay demonstrated that rhein significantly increased the expression of p53 and p21/WAF1 protein, which caused cell cycle arrest. An enhancement in CD95 and its two forms of ligands, membrane-bound CD95 ligand (mCD95L) and soluble CD95 ligand (sCD95L), might be responsible for the apoptotic effect induced by rhein. Taken together, p53 and the CD95/CD95L apoptotic system possibly participated in the antiproliferative activity of rhein in Hep G2 cells. PMID- 14765287 TI - Suppression of infection-induced endotoxin shock in mice by a citrus flavanone naringin. AB - The protective effect of the Citrus flavanone naringin was demonstrated in an endotoxin shock model based on Salmonella infection. Intraperitoneal ( i. p.) infection with 10 (8) CFU Salmonella typhimurium aroA caused lethal shock in lipopolysaccharide (LPS) -responder but not LPS-non-responder mice. Administration of 1 mg naringin 3 h before infection resulted in protection from lethal shock, similar to LPS-non-responder mice. The protective effect of naringin was time- and dose-dependent. Treatment with naringin resulted not only in a significant decrease in bacterial numbers in spleens and livers, but also in a decrease in plasma LPS levels. In addition, naringin markedly suppressed TNF alpha and normalized the activated states of blood coagulation factors such as prothrombin time, fibrinogen concentration and platelet numbers caused by infection. Interestingly, treatment with naringin suppressed high levels of soluble CD14 and high mobility group-1 molecule caused by infection. PMID- 14765288 TI - Impairment of vascular function of rat thoracic aorta in an endothelium-dependent manner by shikonin/alkannin and derivatives isolated from roots of Macrotomia euchroma. AB - The effects of a naphthoquinone analogue, shikonin/alkannin (SA) and derivatives (acetylshikonin and beta,beta-dimethylacrylshikonin), on vascular reactivity were studied with isolated rat aortic rings. At lower concentrations, SA and its derivatives concentration-dependently inhibit the agonist-induced (acetylcholine and histamine) relaxation in PE precontracted aorta in an endothelium-dependent manner with IC (50) values ranging from 0.2 to 1.5 microM. In addition to the effect on agonist-induced vasorelaxation, the Ca (2+) ionophore A23187-induced vasorelaxation was also inhibited or reversed by SA. However, SA had no effect on sodium nitroprusside-induced (guanylate cyclase activator) vasorelaxation. These data suggested that SA and its derivatives might be acting as inhibitors of nitric oxide synthesis in endothelium. At a concentration greater than 10 microM, SA induced contraction of intact but not denuded aorta which could be inhibited by prior treatment with indomethacin, a cyclooxygenase inhibitor. In summary, the results from this study showed that SA and its derivatives inhibited agonist induced relaxation at lower concentrations and induced vasocontraction at higher concentrations. All the effects seen with SA were endothelium-dependent, however, through different mechanisms. Abbreviations. SA:shikonin/alkannin PE:phenylephrine Ach:acetylcholine SNP:sodium nitroprusside eNOS:endothelial nitric oxide synthase L-NAME: Nw-nitro- L-arginine methyl ester PMID- 14765289 TI - Hepatoprotective effect of taxiresinol and (7'R)-7'-hydroxylariciresinol on D galactosamine and lipopolysaccharide-induced liver injury in mice. AB - The hepatoprotective effect of taxiresinol ( 1) and (7' R)-7' hydroxylariciresinol ( 2), two tetrahydrofuran-type lignans isolated from the wood of Taxus yunnanensis, were investigated on D-galactosamine ( D GalN)/lipopolysaccharide (LPS)-induced hepatic liver injury in mice. Pre administration of 1 or 2 at doses of 50 and 10 mg/kg ( i. p.) at 12 and 1 h before D-GalN/LPS injection significantly inhibited hepatocyte DNA fragmentation and apoptotic body formation. Pre-treatment of these two lignans further suppressed hepatic necrosis which occur at later stage of D-GalN/LPS intoxication as demonstrated by the significant and dose-dependent reduction in serum glutamic pyruvic transaminase (sGPT) and serum glutamic oxaloacetic transaminase (sGOT) at 8 h after intoxication. The elevation of serum tumor necrosis factor-alpha (TNF- alpha) level by D-GalN/LPS toxication was significantly inhibited by 1 or 2 at doses of 50 and 10 mg/kg. Moreover, both of these lignans significantly protected hepatocytes from D-GalN/TNF- alpha-induced cell death in primary cultured mouse hepatocytes. These results suggested that 1 and 2 had protected the hepatocytes from apoptosis via an inhibition of TNF- alpha production by activated macrophages and a direct inhibition of apoptosis induced by TNF- alpha in D GalN/LPS-treated mice. PMID- 14765290 TI - Flavonoids and a sesquiterpene lactone from Tanacetum microphyllum inhibit anti inflammatory mediators in LPS-stimulated mouse peritoneal macrophages. AB - In our ongoing research into anti-inflammatory compounds from Tanacetum microphyllum, four naturally occurring flavonoids (santin, ermanin, centaureidin and 5,3'-dihydroxy-4'-methoxy-7-methoxycarbonylflavonol) and one sesquiterpene lactone (hydroxyachillin) isolated from this plant, were evaluated as potential inhibitors of some macrophage functions involved in the inflammatory process. These five compounds significantly inhibited lipopolysaccharide (LPS)-induced nitric oxide (NO) and prostaglandin E (2) (PGE (2)) in a concentration-dependent manner. In the tumour necrosis factor-alpha (TNF-alpha) assay, only centaureidin and hydroxyachillin significantly inhibited the accumulation of TNF-alpha. These results indicate that these compounds may contribute to the anti-inflammatory properties of T. microphyllum. PMID- 14765291 TI - Regulation of gene expression by 8-prenylnaringenin in uterus and liver of Wistar rats. AB - The potential estrogenic activity of 8-prenylnaringenin has been investigated using several in vitro test systems. 8-Prenylnaringenin is a natural secondary product of the female blossoms of hops. The aim of the present study was to characterize 8-prenylnaringenin for its estrogenic effects in vivo. A three day uterotrophic assay was carried out on ovariectomized young female rats. A single dose of 8-prenylnaringenin (10 mg/day/kg body mass) was administered subcutaneously. 17beta-Estradiol (0.03 mg/day/kg body mass; subcutaneous administration) was used as a positive control. Uterine wet weight, endometrial and vaginal epithelial height were determined by histological methods. Gene expression in uterus and in liver was assessed using realtime RT-PCR. Both estradiol and 8-prenylnaringenin significantly stimulated uterine wet weight accompanied by a proliferative response. The three day treatment resulted in a statistically significant increase of the uterine epithelial height as well as of the vaginal epithelial height, the latter being the more sensitive parameter. In the uterus of ovariectomized animals estrogen receptor-alpha and clusterin gene expression were down regulated following treatment with estradiol, whereas expression of complement C3 was up-regulated. In response to treatment with 8 prenylnaringenin the same gene expression pattern was detectable, but less pronounced. The levels of estrogen receptor-alpha mRNA in rat liver were very low and therefore could not be quantitatively assessed. Like in the uterine tissue, estradiol down regulated clusterin expression. The response to 8-prenylnaringenin was weaker but still significant. Conversely, 8-prenylnaringenin was found to be more potent than estradiol in inducing expression of IGFBP-1. In summary, the multiparametric assessment of the estrogenic activity of 8-prenylnaringenin provides overwhelming evidence that 8-prenylnaringenin has largely to be regarded as a pure estrogen agonist and is therefore a questionable candidate molecule for hormone replacement therapy. PMID- 14765292 TI - Rearranged jatrophane-type diterpenes from euphorbia species. Evaluation of their effects on the reversal of multidrug resistance. AB - The rearranged jatrophane-type diterpenes ( 1 - 3), isolated from the Me (2)CO extracts of Euphorbia portlandica and Euphorbia segetalis, were examined for their effects on multidrug resistance (MDR) in mouse lymphoma cells. Compounds 2 and 3 revealed to be active with the latter being more active than the positive control verapamil, a known resistance modifier. The new compound 1, named portlandicine, was isolated from Euphorbia portlandica and its structure characterised by high-field NMR spectroscopic methods including 2D NMR techniques: COSY, HMQC, HMBC and NOESY. The known diterpene 2, together with aleuritolic acid ( 4), oleanolic acid ( 5), and betulin diacetate ( 6), were also isolated from the same species. PMID- 14765293 TI - Phenylpropanoid glycosides from Orobanche caerulescens. AB - Two new phenylpropanoid glycosides, caerulescenoside ( 1), and 3'-methyl crenatoside ( 2), as well as five known phenylpropanoid glycosides [acteoside ( 3), isoacteoside ( 4), campneoside II ( 5), crenatoside ( 6), and desrhamnosyl acteoside ( 7)] were isolated from the whole plant of Orobanche caerulescens. The antioxidative effects of compounds 1 - 7 on human low-density lipoprotein were evaluated. All these compounds suppress concentration-dependently conjugated diene formation with IC (50) values of 1.25 +/- 0.06, 2.97 +/- 0.31, 0.31 +/- 0.01, 1.01 +/- 0.05, 1.15 +/- 0.04, 1.69 +/- 0.15, and 0.64 +/- 0.03 microM, respectively. Comparison of their antioxidative activities with that of resveratrol (IC (50) : 6.75 +/- 1.05 microM), a natural phenolic antioxidant isolated from grape, demonstrated that the prolonged effect on lag-time and the damping effect on oxidative rate by compounds 1 - 7 were all more potent. PMID- 14765294 TI - New diarylheptanoids from the rhizomes of Dioscorea spongiosa and their antiosteoporotic activity. AB - Bioassay-guided fractionation of the water extract of the rhizomes of Dioscorea spongiosa led to the isolation and identification of new diarylheptanoids, diospongins A - C, together with three known lignans. Their structures, including absolute stereochemistry, were determined by analyses of NMR data, chemical conversions and CD spectrum. The isolated compounds, except for diospongin A, exerted potent inhibitory activities on bone resorption induced by parathyroid hormone in a bone organ culture system. PMID- 14765295 TI - 18-Acetoxywithanolides from Physalis chenopodifolia1. AB - Five new withasteroids, physachenolides A - E were isolated from the acetone and methanol extracts of leaves, flowers and stems of Physalis chenopodifolia Lam. The structural assignments of these compounds were based on spectral evidence. The absolute configuration of physachenolide A was established by X-ray analysis. PMID- 14765296 TI - Isotamarixen - a new antioxidant and prolyl endopeptidase-inhibiting triterpenoid from Tamarix hispida. AB - A new pentacyclic triterpenoid, 3alpha-(3",4"-dihydroxy- trans-cinnamoyloxy)- D friedoolean-14-en-28-oic acid ( 1) has been isolated along with two known compounds, rhamnocitrin ( 2) and isorhamnetin ( 3) from the aerial parts of Tamarix hispida Willd. Compound 1 was found to be a potent antioxidant. In addition, compounds 1 - 3 showed significant inhibitory activity against prolylendopeptidase (PEP). PMID- 14765297 TI - Taraxer-14-en-3beta-ol, an anti-inflammatory compound from Sterculia foetida L. AB - Taraxer-14-en-3beta-ol ( 1) was shown to be the active ingredient in the leaves of Sterculia foetida L. The alcohol 1, its acetate and ketone showed anti inflammatory activity against TPA induced mouse ear oedema with inhibition ratios of 60.0, 58.57 and 40.57 at 0.5 mg/ear, respectively. The percentage inhibition of inflammation increased with dose for each compound. PMID- 14765298 TI - In vitro antiprotozoal and cytotoxic activities of some alkaloids, quinones, flavonoids, and coumarins. PMID- 14765299 TI - Chrysopentamine, an antiplasmodial anhydronium base from Strychnos usambarensis leaves. AB - A new derivative of strychnopentamine was isolated from the leaves of Strychnos usambarensis. This compound, named chrysopentamine, was identified by detailed spectroscopic methods (UV, IR, HR-ESI-MS, 1D and 2D NMR). Chrysopentamine presented an original hydroxy substitution on C-14 and an aromatization of the ring D of strychnopentamine leading to anhydronium base properties and exhibited strong antiplasmodial properties (IC (50) less than 1 microM). PMID- 14765300 TI - In vitro antimalarial activity of biflavonoids from Wikstroemia indica. AB - In our investigation of in vitro antimalarial screening of medicinal herbal extracts, the n-BuOH extract from the root of Wikstroemia indica showed a potent inhibitory effect. Fractionation of the active extract led to the isolation of two biflavonoids, sikokianin B ( 1) and sikokianin C ( 2) with IC (50) values 0.54 microg/mL and 0.56 microg/mL, respectively, against the chloroquine resistant strain of Plasmodium falciparum. This is the first report of the biological activity of 1 and 2. As the structure of l has remained unsettled, we confirmed the conformation by (1)H- and (13)C-NMR. PMID- 14765301 TI - Neuroprotective lignans from the bark of Machilus thunbergii. AB - The CH (2)Cl (2) fraction of the bark of Machilus thunbergii Sieb. et Zucc. (Lauraceae) significantly protected primary cultures of rat cortical cells exposed to the excitotoxic amino acid, L-glutamate. (-)-Isoguaiacin, meso dihydroguaiaretic acid, machilin A, (+)-galbelgin, licarin A, (-)-sesamin, and (+)-guaiacin were isolated by bioactivity-guided fractionation from the CH (2)Cl (2) fraction and further separated using chromatographic techniques. Isoguaiacin, meso-dihydroguaiaretic acid, licarin A and (+)-guaiacin had significant neuroprotective activities against glutamate-induced neurotoxicity in primary cultures of rat cortical cells at concentrations ranging from 0.1 microM to 10.0 microM. PMID- 14765302 TI - Pubescenes, jatrophane diterpenes, from Euphorbia pubescens, with multidrug resistance reversing activity on mouse lymphoma cells. AB - The macrocyclic jatrophane diterpene polyesters, pubescenes A - D ( 1 - 4) were isolated from the whole dried plant of Euphorbia pubescens, and evaluated for multidrug resistance (MDR) reversing activity on mouse lymphoma cells. All the compounds displayed very strong activity compared with the positive control verapamil. Pubescene D ( 4) is a new compound, whose structure was established as 3beta,9alpha,-diacetoxy-7beta-benzoyloxy-15beta-hydroxy-14-oxo-2beta H-jatropha-5 E,12 E-diene by spectroscopic methods, including (1)H- (1)H COSY, HMQC, HMBC and NOESY. PMID- 14765303 TI - Geranyl N-dimethylallylanthranilate, a new compound from Esenbeckia yaaxhokob. AB - Through a bioactivity-guided fractionation from the acetone extract of the leaves from Esenbeckia yaaxhokob geranyl N-dimethylallylanthranilate ( 1), the first natural N- and O-prenylated anthranilate, was isolated in addition of the known natural products caryophyllene beta-oxide, caryolane-5beta,9beta-diol, spathulenol ( 2), friedeline, friedelanol, decaprenol, flindersiamine ( 3) and beta-sitosterol. The antimicrobial activities of the extract, fractions and pure compounds were evaluated against two Gram (+) and four Gram (-) bacteria. Compounds 1 and 2 exhibited moderated antimicrobial activity against Staphyloccocus aureus, while 3 was active against S. aureus and Streptococcus faecalis. PMID- 14765304 TI - New halimane diterpenoids from Croton oblongifolius. AB - Three new halimane-type diterpenoids, crotohalimaneic acid ( 1), crotohalimoneic acid ( 2) and 12-benzoyloxycrotohalimaneic acid ( 3), were isolated from the stem bark of Croton oblongifolius Roxb. Their structures were established on the basis of spectroscopic and X-ray analysis. Compounds 1 and 2 showed non-specific strong cytotoxicity against a panel of human tumor cell lines; whereas 3 was inactive. PMID- 14765305 TI - Antifungal steroid saponins from Dioscorea cayenensis. AB - From the rhizomes of Dioscorea cayenensis Lam.-Holl (Dioscoreaceae), the new 26- O- beta- D-glucopyranosyl-22-methoxy-3 beta,26-dihydroxy-25( R)-furost-5-en-3- O- alpha- L-rhamnopyranosyl-(1-->4)- alpha- L-rhamnopyranosyl-(1-->4)-[ alpha- L rhamnopyranosyl-(1-->2)]- beta- D-glucopyranoside ( 1) was isolated together with the known dioscin ( 2) and diosgenin 3- O- alpha- L-rhamnopyranosyl-(1-->4)- alpha- L-rhamnopyranosyl-(1-->4)-[ alpha- L-rhamnopyranosyl-(1-->2)]- beta- D glucopyranoside ( 3). Their structures were established on the basis of spectral data. Compound 2 exhibited antifungal activity against the human pathogenic yeasts Candida albicans, C. glabrata and C. tropicalis (MICs of 12.5, 12.5 and 25 micro g/mL, respectively) whereas 3 showed weak activity and 1 was inactive. PMID- 14765307 TI - Gastroesophageal reflux disease and Barrett's esophagus. AB - Gastroesophageal reflux disease (GERD) is a very common disorder, mainly occurring in Western countries. The nonerosive form of GERD, which occurs in more than half of the patients affected, deserves particular attention. Administering symptomatic therapy without a prior endoscopic examination has become an attractive option, since it also provides diagnostic information. Proton-pump inhibitors (PPIs) have become established as the standard therapy, but new insights into the pathophysiology of the condition may lead to new treatment options using gamma-aminobutyric acid (GABA) agonists. Endoscopic therapy is still at the experimental stage and has yet to prove its value as an alternative to PPI and surgery. However, it is questionable whether antireflux surgery is more cost-effective in the longer term.[nl]Gastroenterologists are now much more aware of Barrett's esophagus than was the case a few years ago. Barrett's esophagus is a frequent finding in patients with reflux symptoms, but is a rare cause of death in affected patients. For several reasons, there is a large gap between recommendations regarding surveillance, on the one hand, and everyday practice on the other. New diagnostic procedures such as chromoendoscopy may allow better detection of premalignant and malignant alterations in metaplastic mucosa, but the safety of such techniques has been questioned. Prophylactic ablation is a debatable approach, whereas endoscopic interventions in patients with high-grade dysplasia and early adenocarcinoma are continuing to develop as attractive alternatives to esophagectomy in selected patients. It remains to be seen whether chemoprevention using cyclooxygenase-2 (COX-2) inhibitors should be carried out in high-risk patients with Barrett's esophagus, in order to prevent malignant transformation to esophageal cancer. PMID- 14765308 TI - Diagnosis of esophagogastric tumors. AB - Esophagogastric tumors occur in three sectors: the esophagus, the EG junction and the non-cardia stomach. Neoplasia develops in the squamous stratified epithelium of the esophagus and in the columnar epithelium of the Barrett's esophagus or in the stomach. At the junction, tumors arise either in a very short Barrett's esophagus or in the gastric epithelium of the cardia. The prognosis of tumors detected at the advanced stage is poor. Secondary prevention requires detection at the early stage. Most superficial neoplastic lesions in the esophagus and in the stomach have a non-protruding appearance, which is similar for premalignant and malignant lesions. Improved accuracy in endoscopic diagnosis and prediction of histology prior to biopsy and treatment decision is based upon magnification with a optical zoom and electronic processing of the captured image with structure enhancement, enhancement of the color of hemoglobin and narrow band imaging. This applies particularly to the exploration of the Barrett's esophagus for identification of the areas with intestinal metaplasia and of flat neoplastic areas. In spite of the predictive value of endoscopy for histology, biopsy samples are still required for pathology and eventually studies with biological markers. Spectroscopic techniques provide a new perspective, up to the level of molecular endoscopy, but they are unlikely to be cost/effective. The classification in the sub-types 0 of neoplastic lesions has some relevance to prediction of depth of invasion. In the esophagus, EUS staging with high frequency miniprobes is a useful complement. PMID- 14765309 TI - Variceal bleeding and portal hypertension: new lights on old horizon. AB - New clinical, endoscopic, and imaging modalities for diagnosing varices and predicting bleeding are being investigated. Transnasal endoscopy and ultrathin battery-powered esophagoscopes are being used to improve patient comfort and compliance. Patients who respond to portal pressure-reducing drugs not only have a reduced risk of bleeding, but also a reduced risk of developing other complications, with improved survival. Nitrates have been shown to have no definite role in primary prophylaxis against variceal bleeding. The hemodynamic response to treatment has an independent prognostic value for the risk of variceal bleeding. Newer drugs have been investigated for reducing the hepatic venous pressure gradient, but with little success. Survival after bleeding has increased due to improved patient care and technological advances. Combined radiographic and endoscopic management of gastric varices is evolving and appears to be promising. Nonvariceal bleeding from portal hypertensive gastropathy is increasingly being recognized as a potential cause of bleeding in patients with portal hypertension, and pharmacotherapy with octreotide appears to be promising for the treatment of this condition. Variceal band ligation in children has been found to be as safe and effective as in adults. PMID- 14765310 TI - Inflammatory bowel disease: what is new? AB - This article reviews a selection of 50 papers on inflammatory bowel disease published between April 2002 and August 2003. The new information and progress summarized here includes: the role of bacteria, their interaction with CARD15/NOD2 gene products and the modulating effects of probiotics; new imaging techniques, such as wireless capsule endoscopy and chromoendoscopy for detecting dysplasia, and fecal markers. It has not been possible to review every aspect of treatment, and the focus here is on factors capable of predicting the efficacy and tolerance of infliximab and new treatments, including natalizumab, thalidomide, cyclophosphamide and 6-thioguanine. PMID- 14765311 TI - Diagnostic endoscopic retrograde cholangiopancreatography. PMID- 14765312 TI - Therapeutic biliary endoscopy. AB - Almost all biliary diseases are now amenable to endoscopic treatment. Progress in therapeutic biliary endoscopy is currently focusing on identifying the best treatment options to be used in each case in relation to the available evidence: how should a malignant stricture of the common bile duct or hilum be drained? How should an external biliary fistula be treated? What should one do with the gallbladder after extraction of common bile duct stones? Should sedation or general anesthesia be offered? Progress has been made in the technique of endoscopic ampullectomy and in developing new devices to improve the success rate of endoscopic retrograde cholangiopancreatography. PMID- 14765314 TI - Usefulness of magnifying endoscopy in the diagnosis of early gastric cancer. AB - BACKGROUND AND STUDY AIMS: We investigated the characteristic findings of early gastric cancer revealed by magnifying endoscopy, and clarified their relationship with histopathological features. PATIENTS AND METHODS: A total of 74 patients with early gastric cancer underwent magnifying endoscopy ( x 80) between March 2000 and December 2001. The endoscopic findings demonstrated 11 elevated-type carcinomas and 63 depressed-type, and histological examination showed 56 differentiated carcinomas and 18 undifferentiated carcinomas. The histopathological results were compared with findings from magnifying endoscopy regarding minute surface structure and microvessels. RESULTS: We were able to roughly classify the minute surface structure of early gastric cancer as shown by magnifying endoscopy into three patterns, as follows: (i). a small regular pattern of sulci and ridges; (ii). an irregular pattern of sulci and ridges; and (iii). a lack of visible structure. Abnormal microvessels observed in cancerous lesions were classified according to two patterns: irregular minute vessels and variation of vessel caliber. The small regular pattern of sulci and ridges was significantly more frequently observed in differentiated carcinoma (30/56, 53.6 %) than in undifferentiated carcinoma (2/18, 11.1 %). Lack of visible structure and irregular minute vessels were significantly more frequently observed in undifferentiated carcinoma (44.4 % and 77.7 %) than in differentiated carcinomas (5.4 % and 51.8 %). CONCLUSION: The minute surface structure and microvessels observed by magnifying endoscopy were related to histopathological findings. Magnifying endoscopy is valuable for predicting the histological nature in the diagnosis of early gastric cancer. PMID- 14765313 TI - Inter- and intra-observer variability of magnification chromoendoscopy for detecting specialized intestinal metaplasia at the gastroesophageal junction. AB - BACKGROUND AND STUDY AIMS: Magnification endoscopy after contrast enhancement with acetic acid or staining with methylene blue has been reported to be highly accurate in predicting specialized intestinal metaplasia (SIM) in Barrett's esophagus. So far, however, no data have been published on the interobserver and intra-observer variability of these new methods. PATIENTS AND METHODS: Fifty-one patients with reflux symptoms were prospectively evaluated. Endoscopy was carried out with a magnification endoscope, and video sequences were recorded in standard and zoom modes (at the 12-o'clock, 3-o'clock, 6-o'clock, and 9-o'clock positions) before and after instillation of 1.5 % acetic acid (n = 26) or staining with 0.5 % methylene blue (n = 25). Biopsies were obtained from the same locations for histopathological examination. The 102 video sequences were shown to four experienced endoscopists in a mixed and blinded fashion. The evaluation criteria used followed the published criteria; classification was carried out according to the pit-pattern structure, methylene blue positivity, and the presence of villous structures. Finally, a general statement on suspected SIM in relation to Barrett's esophagus was requested. RESULTS: With regard to the criteria selected for evaluation, there was a high level of interobserver variability among the four examiners (all kappa < 0.4). SIM was histologically detectable in 60.8 % of the patients. The accuracy of all of the examiners for predicting SIM by magnification endoscopy was around 50 %, with no differences observed before and after instillation of acetic acid or methylene blue staining. CONCLUSIONS: The suggested criteria for identifying SIM using magnification endoscopy are associated with a high level of interobserver variability. When evaluated in a blinded manner, staining techniques do not significantly improve the yield for detecting SIM at the esophagogastric junction. PMID- 14765315 TI - Botulinum toxin injection after biliary sphincterotomy. AB - BACKGROUND AND STUDY AIMS: Endoscopic biliary sphincterotomy in patients with sphincter of Oddi dysfunction (SOD) is associated with a high risk of pancreatitis after endoscopic retrograde cholangiopancreatography (ERCP), which may be secondary to residual pancreatic sphincter hypertension. It was hypothesized that botulinum toxin injection could be used to reduce pancreatic sphincter hypertension temporarily in SOD patients after biliary sphincterotomy, thereby reducing the rate of procedure-induced pancreatitis. PATIENTS AND METHODS: All patients undergoing ERCP with manometry due to a suspected biliary SOD were asked to participate in the study. Patients with elevated basal sphincter pressures were randomly assigned to receive either botulinum toxin or a sham saline injection after biliary sphincterotomy. Fifty units of botulinum toxin were delivered via a sclerotherapy needle in the form of two 25-U injections of 0.25 ml each into the pancreatic sphincter. In patients in the sham arm, 0.50 ml of saline was injected into the duodenal lumen. RESULTS: Between 12 February 1999 and 29 November 2000, a total of 98 patients were referred for ERCP with manometry; 86 consented to participate in the study, and 26 had elevated baseline pressures and underwent random assignment. Twelve received botulinum toxin injection and 14 were randomly assigned to receive the sham injection. A total of six patients in the sham group (43 %) developed procedure-induced pancreatitis, compared with three patients in the botulinum toxin group (25 %; P = 0.34). CONCLUSIONS: Biliary sphincterotomy in patients with sphincter of Oddi dysfunction without pancreatic protection is risky and should no longer be carried out. This study demonstrates that botulinum toxin injection into the residual pancreatic sphincter after biliary sphincterotomy is technically feasible and safe, showing a trend toward a reduced post-ERCP pancreatitis rate in patients with sphincter of Oddi dysfunction. Further studies will need to confirm the validity of these experimental results before this technique can be used routinely. PMID- 14765316 TI - Suspected sphincter of Oddi dysfunction type II: empirical biliary sphincterotomy or manometry-guided therapy? AB - BACKGROUND AND STUDY AIMS: Sphincter of Oddi manometry is considered to be the gold standard for diagnosing sphincter of Oddi dysfunction (SOD). Elevated basal sphincter pressures are found in about half of the patients with findings consistent with biliary type II SOD, and most of these patients will symptomatically improve after endoscopic sphincterotomy. Since manometric sphincter evaluation is not widely available, a decision analysis was used to compare the overall costs and outcomes of manometry-directed therapy with "empirical" sphincterotomy in patients with suspected biliary type II SOD. PATIENTS AND METHODS: A decision analysis model was constructed using a software program. In a hypothetical cohort of 100 patients with suspected type II SOD, the following strategies were evaluated: a). endoscopic retrograde cholangiopancreatography (ERCP) with manometry followed by biliary sphincterotomy only if an elevated sphincter of Oddi basal pressure was found; and b). "empirical" biliary sphincterotomy without manometry. Data on the probability of an elevated sphincter of Oddi basal pressure at the time of ERCP in patients with suspected biliary SOD type II, the proportion of patients who improved after biliary sphincterotomy (with and without elevated basal pressures), the proportion of patients who improved without biliary sphincterotomy, complications, and death were obtained from the literature and from our center. The procedural and hospitalization costs represented the average Medicare reimbursement at our institution. The expected overall costs and numbers of patients improving with each strategy were compared.[nl] RESULTS: The strategy of ERCP with manometry resulted in total costs of US dollars 2790 per patient, whereas a strategy of "empirical" biliary sphincterotomy resulted in total costs of US dollars 2244. In a cohort of 100 patients with suspected SOD, 55 % of patients would be expected to improve if manometry were performed, compared to 60 % of patients improving with "empirical" biliary sphincterotomy. Univariate sensitivity analyses demonstrated that "empirical" biliary sphincterotomy continued to be a cost-saving strategy in comparison with ERCP with manometry as long as the probability of spontaneous improvement in patients with "normal" manometry was less than 41 %, the probability of complications associated with manometry was greater than 6 %, and the probability of complications due to biliary sphincterotomy was less than 19 %. CONCLUSIONS: For patients with suspected biliary SOD type II, empirical biliary sphincterotomy performed by experienced endoscopists appears to be cost-saving in comparison with a strategy based on the results of manometry. PMID- 14765317 TI - Sphincter of Oddi dysfunction: cut and inject, but don't measure the pressure? PMID- 14765318 TI - Dieulafoy's lesion of the esophagus correctly diagnosed and successfully treated by the endoscopic injection of N-butyl-2-cyanoacrylate. AB - Dieulafoy's lesion is an arterial malformation in the subumucosal layer of the gastrointestinal tract that can cause massive bleeding. The esophagus is not a common location for this lesion. We present here a first report of Dieulafoy's lesion of the esophagus correctly diagnosed and successfully treated by the endoscopic injection of N-butyl-2-cyanoacrylate. PMID- 14765321 TI - Invasive intraductal papillary mucinous tumor of the pancreas with simultaneous invasion of the stomach and duodenum. AB - An 81-year-old woman was admitted with epigastric pain and weight loss. She had been diagnosed with an intraductal papillary mucosal tumor (IPMT) 7 years previously, but had refused surgery for religious reasons. Esophagogastroduodenoscopy revealed a nodular, elevated lesion that was discharging mucin into the duodenal bulb and posterior wall of the upper body of the stomach. Endoscopic ultrasonography, abdominal computed tomography, and endoscopic retrograde cholangiography were carried out, and a highly invasive IPMT with simultaneous invasion of the stomach and duodenum was diagnosed. PMID- 14765322 TI - Tension pneumothorax as a complication of colonoscopy. PMID- 14765323 TI - A new device for better control of the polypectomy snare wire. PMID- 14765324 TI - Treatment of radiation-induced hemorrhagic duodenitis with argon plasma coagulation. PMID- 14765325 TI - Successful removal from the duodenum of swallowed sewing needles using devised endoscopic forceps. PMID- 14765326 TI - High-magnification chromoscopic ileoscopy in familial adenomatous polyposis: detection in vivo of colonic metaplasia and microadenoma formation. PMID- 14765328 TI - [Renal function in hypertensive patients with obstructive sleep apnea]. AB - BACKGROUND: Patients with hypertension often suffer from obstructive sleep apnea (OSA). In addition to hypertension several other risk factors (hypoxemia, hyperlipidemia, and increased sympathic nerve activity) may contribute to progressive renal dysfunction in OSA patients. The aim of this study was to compare renal function in OSA-patients with and without hypertension. METHODS: 81 consecutive patients (50 males, 31 females) were screened for sleep apnea. Parameters of renal function (serum creatinine, creatinine clearance, microalbuminuria), and of lipid and glucose metabolism were correlated to polysomnographic results. RESULTS: OSA (apnea/hypopnea index [AHI] > or = 5) was found in 57 of 81 patients. Mean AHI was 26.7 +/- 26.1. Hypertension (blood pressure > or = 140/90 mmHg or use of antihypertensive drugs) occurred in 63 of 81 patients. Serum creatinine in OSA patients was significantly higher than in patients without OSA (1.11 +/- 0.15 vs. 0.91 +/- 0.12 mg/dl, p < 0.001). Serum creatinine correlated significantly with AHI. Creatinine clearance was associated with age (r = -0.314; p = 0.014) and presence of OSA (r = 0.265; p = 0.093). No correlation was shown between hypertension and serum creatinine or creatinine clearance. Microalbuminuria was not associated with OSA. CONCLUSION: Our results suggest an independent association between OSA and impaired renal function. Further prospective studies will have to be done to elucidate the pathophysiological mechanisms. PMID- 14765327 TI - [Factors influencing duration of hospitalization after stroke in Germany]. AB - BACKGROUND: In Germany up-to-date data within community settings about factors influencing length of stay in acute hospitals are lacking. We, therefore, identified predictors for length of stay in acute hospital after ischemic stroke in a pooled analysis of large German stroke registers. METHODS: Ischemic stroke patients admitted to hospitals cooperating within the German Stroke Registers Study Group (ADSR) between January 1, 2000 and December 31, 2000 were analysed. The influence of patients' demographic and clinical characteristics as well as the characteristics of the treating hospitals on length of stay were analysed by multivariate linear regression. RESULTS: Overall, 13 440 patients after ischemic stroke were included in the analyses. Their mean age was 70 years, 53 % were men. Median length of stay in acute hospitals was 12 days. In multivariate analyses younger age, an increasing number of co-morbidities, and an increasing number of neurological deficits were identified as predictors of prolonged stay in hospital. Patients were more likely to stay longer in an acute hospital if they were to be discharged to a rehabilitation unit or to a nursing home. Length of stay was independently decreased for patients treated in hospitals providing acute stroke unit services and for hospitals treating more than 250 stroke patients per year. CONCLUSION: In addition to patients' demographic and clinical characteristics, length of stay in hospital was influenced by the hospitals' characteristics. Especially the volume of treated patients and the organisation of services within the hospital may play the key role. PMID- 14765329 TI - [Hypophyseal ACTH-cell carcinoma after several surgical interventions and radiotherapy]. AB - HISTORY AND ADMISSION FINDINGS: A 56-year-old woman was admitted to our hospital with headache, especially on the left temporal side, dizziness and exercise intolerance. She had been operated three times and radiotherapy once because of pituitary adenoma with intermittend hypercortisolism. The clinical examination was without abnormal findings apart from left temporal pain on pressure on the top of the skull. INVESTIGATIONS: Blood tests were entirely normal. At cranial magnet resonance imaging (cMRI) a left temporal tumor of 10 mm diameter was diagnosed. TREATMENT AND COURSE: The first histological study of the excized lesion could not clarify the diagnosis completely. Because of a local recurrent tumor of 20 mm, a second operation was necessary two months later. Due to structural and immunohistological similarities this tumor was identified as a metastasis of a pituitary ACTH-cell carcinoma. The patient was given adjuvant stereotactic radiotherapy. Two years after the treatment, no tumor recurrence was seen by cMRI. CONCLUSION: Carcinomas of the pituitary are very rare. They can be diagnosed only by their metastases. The pathogenesis is still unclear. It is debatable, whether surgery and/or X-ray therapy in the past may influence tumor development. PMID- 14765330 TI - [Repeated improper intravenous injection of fentanyl from a transdermal system]. AB - We report on a 30 year-old male who misused transdermal fentanyl. He injected the contents of transdermal patches intravenously. Suffering from chronic pain following total hip replacement, he had received a prescription for this drug formulation from his general practitioner. During his stay in a pain clinic he was able to obtain a total of 13 fentanyl patches from other patients or local pharmacies. He became seriously ill with multiple organ dysfunction in the course of an infection of his thigh. After surgical and intensive care treatment he recovered soon, but the hip prosthesis had to be explanted. There are some reports in the literature of misuse of fentanyl patches. The contents may be ingested orally, or they can be inhaled. Aspirated with a syringe the content of fentanyl patches can also be injected intravenously, sometimes resulting in exit us. Prescribers must be aware of the potential for abuse of fentanyl patches which can be stolen, sold or even removed from dead bodies. PMID- 14765331 TI - [Rehabilitation after ST-elevation myocardial infarction: inpatient, outpatient or none?]. PMID- 14765332 TI - [Is the supplementation with antioxidants effective in the treatment of atherosclerosis?]. AB - Results from experimental and epidemiologic studies suggest an influence of oxidative stress on development and progression of atherosclerosis in man. Prospective endpoint studies failed to support this hypothesis. The current literature is summarized in this review. PMID- 14765333 TI - [Legal liability aspects of physician's drug prescription and drug use]. PMID- 14765334 TI - [Underutilization of cardiovascular pharmacotherapy: how to pay for it?]. PMID- 14765335 TI - [Life style and health care costs]. PMID- 14765336 TI - [Can a patient do heavy physical work after myocardial infarction? Concerning the article in DMW 40/2003]. PMID- 14765337 TI - Viral replication in the nasopharynx is associated with diarrhea in patients with severe acute respiratory syndrome. AB - The role of severe acute respiratory syndrome (SARS) coronavirus as an enteric pathogen was investigated in a cohort of 142 patients with SARS who were treated with a standard treatment protocol. Data from daily hematological, biochemical, radiological, and microbiological investigations were prospectively collected, and the correlation of these findings with diarrhea was retrospectively analyzed. Sixty-nine patients (48.6%) developed diarrhea at a mean (+/- standard deviation [SD]) of 7.6+/-2.6 days after the onset of symptoms. The diarrhea was most severe at a mean (+/-SD) of 8.8+/-2.4 days after onset, with a maximum frequency of 24 episodes per day (median, 5 episodes; range, 3-24 episodes). A higher mean virus load in nasopharyngeal specimens obtained on day 10 after the onset of symptoms was significantly associated with the occurrence of diarrhea (3.1 log10 vs. 1.8 log10 copies/mL; P=.01) and mortality (6.2 vs. 1.7 log10 copies/mL; P<.01). However, diarrhea was not associated with mortality. The lung and the gastrointestinal tract may react differently to SARS coronavirus infection. Additional investigation of the role of SARS coronavirus in the pathogenesis of diarrhea in patients with SARS should be conducted. PMID- 14765338 TI - Acceptable rates of treatment failure in osteomyelitis involving the diabetic foot: a survey of infectious diseases consultants. AB - Shortening the duration of antibiotic therapy is an attractive strategy for delaying the emergence of antimicrobial resistance. The paucity of data about optimal treatment durations hinders adoption of this approach. This study used contingent valuation analysis to identify failure rates for treatment of diabetic foot osteomyelitis acceptable to infectious diseases consultants (IDCs). The Infectious Diseases Society of America's Emerging Infections Network (EIN) provided members with the case scenario and 1 of 10 failure rates; members were asked, assuming delivery of standard therapy, if they would accept or reject the given failure rate. The relationship between specific failure rates and the willingness of IDCs to accept them was analyzed. The median acceptable failure rate for EIN members was 18.1%; 75% of IDCs found a failure rate of 7.8% to be acceptable, and 25% found a rate of 28.4% to be acceptable. The methodology used in this study may prove useful in delineating acceptable treatment failure thresholds, an initial step in shortening durations of antimicrobial therapy. PMID- 14765340 TI - Predicting relapse after cessation of Lamivudine monotherapy for chronic hepatitis B virus infection. AB - There have been reports of relapse after cessation of lamivudine monotherapy for hepatitis B virus (HBV) infection. The aim of this study was to examine factors that predict posttreatment relapse. Comparison 22 patients who experienced relapse with 11 who did not after cessation of therapy showed that predictive factors for nonrelapse were hepatitis B e antigen seroconversion and duration of undetectable HBV DNA load (<0.7 log IU/mL), as determined by HBV real-time detection direct testing. However, 7 of 12 patients with seroconversion experienced relapse after cessation of therapy. Multivariate analysis revealed that the duration of an undetectable HBV DNA load was the only independent predictive factor for nonrelapse (odds ratio, 0.50; 95% confidence interval, 0.27 0.9). More-prolonged lamivudine therapy is required after seroconversion, and persistent duration of an HBV DNA level of <0.7 log IU/mL for >6 months can more accurately aid in the decision of when to stop lamivudine therapy. PMID- 14765339 TI - Prognostic factors for severe acute respiratory syndrome: a clinical analysis of 165 cases. AB - This study analyzes single factors that affect the prognosis of severe acute respiratory syndrome (SARS) and establishes a prognosis model by multivariate analysis. We retrospectively analyzed the clinical features of SARS in 165 clinically confirmed severe cases. Both age and existence of other diseases before SARS were significantly correlated with prognosis (r=0.506 and r=0.457, respectively; P<.001). During the acute phase of SARS, lactate dehydrogenase level, degree of hypoxemia, respiratory rate, alpha -hydroxybutyric dehydrogenase level, creatine kinase isoenzyme-MB, platelet count, and number of involved lobes noted on chest radiographs, and so on, correlated markedly with the prognosis (r=0.257-0.788; P<.05). The multivariate prognosis regression model was associated with degree of hypoxemia and platelet count. The model was defined by the formula Py=1=es/(1+es), where S is [2.490 x degree of hypoxemia]-[0.050 x number of platelets], and it had a high sensitivity (91.67%), specificity (98.33%), and accuracy (96.42%). The model could be used to effectively judge the state of illness and the prognosis. PMID- 14765341 TI - Invasive pneumococcal disease among Navajo adults, 1989-1998. AB - Compared with white and black persons in the United States, some Native American groups are at increased risk for invasive pneumococcal disease (IPD). To characterize the epidemiology of IPD among Navajo adults, we conducted active surveillance for IPD on the Navajo Nation and reviewed medical records of patients with IPD. For 1997-1998, the annual incidence (cases per 100,000 persons) was 56 for Navajos aged 18-64 years and 190 for Navajos aged > or =65 years. The corresponding rates were 10 and 57 for white and 44 and 82 for black persons in the United States. The case-fatality rate was 14%. Eighty percent of cases were caused by serotypes included in the 23-valent pneumococcal polysaccharide vaccine. Navajo adults have rates of IPD that are 3-5-fold higher than those of the general US population. Additional research is needed to understand the reasons for this elevated risk and to develop prevention strategies. PMID- 14765342 TI - Determination of serum antibody to Bordetella pertussis adenylate cyclase toxin in vaccinated and unvaccinated children and in children and adults with pertussis. AB - Presence of antibody to adenylate cyclase toxin (ACT) has been noted following Bordetella pertussis infection. Because ACT is not presently in any acellular pertussis vaccines, it has been considered as a possible antigen to use in B. pertussis diagnostic enzyme-linked immunosorbent assay (ELISA) studies. We determined antibody to B. pertussis ACT by ELISA and Western blot tests in serum samples obtained from unvaccinated children, from children vaccinated with several diphtheria and tetanus toxoid vaccines (DTP vaccines), from children vaccinated with vaccines containing acellular pertussis components in combination with diphtheria and tetanus toxoids (DTaP vaccines), and from children and adults with pertussis. Primary infections with either B. pertussis or Bordetella parapertussis stimulated a vigorous antibody response to ACT. In contrast, patients in whom DTP and DTaP vaccines failed had minimal ACT antibody responses. The lack of a significant ACT antibody response in children in whom the vaccine failed is of interest but would seem to preclude the use of ACT in diagnostic tests. PMID- 14765343 TI - High-level penicillin-nonsusceptible Streptococcus pneumoniae bacteremia: identification of a low-risk subgroup. AB - High-level penicillin resistance has been associated with treatment failure in patients with Streptococcus pneumoniae infections. To identify a subgroup of patients at low risk for high-level penicillin-nonsusceptible S. pneumoniae bacteremia, a cross-sectional study of 303 patients was performed. For the total study population, penicillin resistance was observed in 98 (32%) of 303 patients; high-level resistance was seen in 33 (11%). A predictive model was created by using 3 baseline variables that were independently associated with high-level penicillin resistance: previous beta -lactam antibiotic use, previous stay in a risk area (defined as stay in day care facilities, prisons, homeless shelters, nursing homes, or other long-term care facilities), and previous respiratory tract infection. The model was used to identify patients at low and high risk for high-level penicillin-resistant pneumococcal bacteremia. None of the isolates of patients in the low-risk subgroup had ceftriaxone resistance. Patients in the low risk subgroup could be empirically treated with fluoroquinolone-sparing regimens. PMID- 14765344 TI - Unusual presentation of life-threatening Toscana virus meningoencephalitis. AB - This case report describes a brother and a sister with severe meningoencephalitis caused by Toscana virus (TOSv). The clinical presentation was characterized by stiff neck, deep coma, maculopapular rash, diffuse lymphadenopathy, hepatosplenomegaly, renal involvement, tendency to bleeding, and diffuse intravascular coagulation. The boy had epididymo-orchitis. Recovery with neurologic sequelae as hydrocephalus was observed. Microbiological diagnosis was obtained by serological tests and reverse transcriptase-polymerase chain reaction. Sequencing of polymerase chain reaction products from the S and M segments was carried out. TOSv may be a causative agent in severe meningoencephalitis. PMID- 14765345 TI - Treatment outcomes for serious infections caused by methicillin-resistant Staphylococcus aureus with reduced vancomycin susceptibility. AB - Although infections caused by methicillin-resistant Staphylococcus aureus with reduced vancomycin susceptibility (SA-RVS) have been reported from a number of countries, including Australia, the optimal therapy is unknown. We reviewed the clinical features, therapy, and outcome of 25 patients with serious infections due to SA-RVS in Australia and New Zealand. Eight patients had endocarditis, 9 had bacteremia associated with deep-seated infection, 6 had osteomyelitis or septic arthritis, and 2 had empyema. All patients had received vancomycin before the isolation of SA-RVS, and glycopeptide treatment had failed for 19 patients (76%). Twenty-one patients subsequently received active treatment, which was effective for 16 patients (76%). Eighteen patients received linezolid, which was effective in 14 (78%), including 4 patients with endocarditis. Twelve patients received a combination of rifampicin and fusidic acid. Surgical intervention was required for 15 patients (60%). Antibiotic therapy, especially linezolid with or without rifampicin and fusidic acid, in conjunction with surgical debulking is effective therapy for the majority of patients with serious infections (including endocarditis) caused by SA-RVS. PMID- 14765346 TI - Cat-transmitted sporotrichosis epidemic in Rio de Janeiro, Brazil: description of a series of cases. AB - Sporotrichosis is the most common subcutaneous mycosis in South America. Classic infection is associated with traumatic inoculation of soil, vegetables, and organic matter contaminated with Sporothrix schenckii. Zoonotic transmission has been described in isolated cases or in small outbreaks. Since 1998, we have been observing an increasing number of cases of sporotrichosis in persons from the city of Rio de Janeiro, Brazil, and surroundings. From 1998 to 2001, 178 cases of culture-proven sporotrichosis had been diagnosed. Female patients predominated, and the median age was 39 years. The most frequent clinical presentation was lymphocutaneous disease. Of the 178 patients, 156 reported domiciliary or professional contact with cats with sporotrichosis, and 97 of these patients had a history of receipt of cat scratch or bite. The patients received itraconazole as first-line treatment. This study suggests that feline transmission of sporotrichosis was associated with a large and long-lasting outbreak of the disease in Rio de Janeiro. PMID- 14765347 TI - Risk of vaccinia transfer to the hands of vaccinated persons after smallpox immunization. AB - Transmission of vaccinia virus after smallpox vaccination is a concern. We conducted a prospective examination of the protection afforded by vaccination site bandages in recently vaccinated individuals. After smallpox vaccination, inoculation sites were covered with 2 occlusive dressings. Site assessment and bandage changes occurred every 3-5 days until the site was healed. At each visit, specimens from the vaccination site, outer dressing surface, and contralateral hand were obtained for vaccinia culture. For 148 vaccinated subjects, vaccinia was detected from vaccination lesions of every subject on several occasions. Only 6 (0.65%) of 918 dressing (95% CI, 0.24%-1.4%) and 2 (0.22%) of 926 hand (95% CI, 0.03%-0.78%) specimens tested positive for vaccinia. The mean number of bandage changes was 9.6 (95% CI, 9.17-10.0). Vaccinia autoinoculation did not occur. The rate of vaccinia recovery outside occlusive bandages covering smallpox vaccination sites was remarkably low, suggesting excellent protection against inadvertent transmission. PMID- 14765348 TI - Postoperative endophthalmitis. AB - Postoperative endophthalmitis remains a serious clinical problem in ophthalmology, with an incidence of approximately 0.5%. Prognosis is largely determined by the virulence of the offending organism. The Endophthalmitis Vitrectomy Study (EVS) was a prospective, randomized trial comparing various diagnostic and treatment modalities in cases of endophthalmitis that followed cataract surgery. The EVS found that vitrectomy was only beneficial for patients presenting with very poor visual acuity and that intravenous antibiotic treatments had no additional benefit, compared with intravitreal antibiotic therapy alone. However, weaknesses of the EVS leave these conclusions open to modification in the future. Preoperative application of povidone-iodine preparation to the skin and conjunctiva is the only proven endophthalmitis prophylaxis. Endophthalmitis may be chronic and may follow glaucoma surgery and intravitreal injection of gas and drugs. The EVS did not study these issues, although they are associated with specific features that may require alterations in patient management. PMID- 14765349 TI - Infectious disease images on the World Wide Web. AB - Infectious diseases (ID) physicians are often in need of medical images to enhance their teaching, research, and clinical practice. We explored the Internet for Web sites with images that would be useful to ID physicians. A total of 24 sites were included for review. Of these, 10 sites were broad in their scope and included images of bacteria, parasites, viruses, and/or fungi. In addition, 4 sites reviewed were specific for fungi, 4 sites were specific for viruses, and 6 sites focused on parasites. The number and size of images, copyright restrictions, and fees were noted for all sites. The authors gave each site a subjective navigation and layout score. Features of the sites, including microscopy images, laboratory images, clinical images, clinical vignettes, medical illustrations and/or clip art, medical illustrations of life cycles, slides of educational material, radiographs, and animation or video, were also described. A variety of image resources are available to the ID physician. PMID- 14765350 TI - The clinical pharmacokinetics of pyrazinamide in HIV-infected persons with tuberculosis. AB - The pharmacokinetics of pyrazinamide (PZA) in patients with human immunodeficiency virus (HIV)-related tuberculosis are incompletely characterized. Serum PZA concentrations were determined at 2, 6, and 10 h after dosing in 48 subjects with HIV-related tuberculosis. Estimates of drug exposure using 2-h concentrations and 2- and 3-time point estimates of area under time-concentration curves (AUCs) were compared. For daily dosing, 2-h concentrations less than low and very low literature-defined cut points (i.e., 20 and 10 mg/L) were noted for 2 subjects (4%) and 1 subject (2%), respectively. For intermittent PZA dosing, 1 subject (4%) had a 2-h concentration that was less than the low cut point (25 mg/L). Correlations between 2-h concentration and AUC estimates based on 2- or 3 time point concentration determinations were strong. In HIV-infected persons receiving antituberculosis regimens containing PZA, lower-than-expected 2-h concentrations are uncommon. For therapeutic monitoring of PZA drug exposure, determination of a 2-h postdose concentration appears as reliable as 2- or 3-time point estimates of the AUC for PZA. PMID- 14765351 TI - Discontinuation of maintenance therapy for cryptococcal meningitis in patients with AIDS treated with highly active antiretroviral therapy: an international observational study. AB - We conducted a retrospective, multicenter study evaluating the safety of discontinuing maintenance therapy for cryptococcal meningitis after immune reconstitution. Inclusion criteria were a previous definitive diagnosis of cryptococcal meningitis, a CD4 cell count of >100 cells/microL while receiving highly active antiretroviral therapy (HAART), and the subsequent discontinuation of maintenance therapy for cryptococcal meningitis. The primary end point was relapse of cryptococcal disease. As of July 2002, 100 patients were enrolled. When maintenance therapy was discontinued, the median CD4 cell count was 259 cells/microL and the median plasma virus load was <2.30 log10 copies/mL, and serum cryptococcal antigen was undetectable in 56 patients. During a median follow-up period of 28.4 months (range, 6.7-64.5; 262 person-years), 4 events were observed (incidence, 1.53 events per 100 person-years; 95% confidence interval, 0.42-3.92). Three of these patients had a CD4 cell count of >100 cells/microL and a positive serum cryptococcal antigen test result during the recurrent episode. In conclusion, discontinuation of maintenance therapy for cryptococcal meningitis is safe if the CD4 cell count increases to >100 cells/microL while receiving HAART. Recurrent cryptococcal infection should be suspected in patients whose serum cryptococcal antigen test results revert back to positive after discontinuation of maintenance therapy. PMID- 14765352 TI - Health-related quality of life of patients with HIV disease: impact of hepatitis C coinfection. AB - Health-related quality of life (HRQOL) is diminished in patients infected with both hepatitis C virus (HCV) and human immunodeficiency virus (HIV), but the effect of HIV/HCV coinfection on HRQOL is unknown. We compared the HRQOL of urban HIV/HCV coinfected patients with that of patients infected with either HCV or HIV alone. We then compared the 3 groups with a US population sample, adjusting for demographic characteristics. HRQOL for the group of HIV/HCV coinfected patients was statistically similar to that of HRQOL in patients with either HCV or HIV alone, but the 3 groups had a significantly decreased HRQOL than did the adjusted US population. Using multivariate techniques, we determined that age, unemployment, injection drug use, and depression were associated with impaired HRQOL. These findings underscore the importance of a multidisciplinary approach to the treatment of these patient populations. PMID- 14765354 TI - Severe acute respiratory syndrome without respiratory symptoms or abnormal chest radiograph findings. AB - We report a serologically confirmed case of severe acute respiratory syndrome (SARS) in a 27-year-old health care worker. The patient reported no respiratory complaints, and the findings of serial chest radiographs were normal. This case illustrates the wide spectrum of clinical illness caused by infection with SARS coronavirus. PMID- 14765353 TI - The effect of highly active antiretroviral therapy on dermatologic disease in a longitudinal study of HIV type 1-infected women. AB - The effect of highly active antiretroviral therapy (HAART) on skin diseases was evaluated in 878 human immunodeficiency virus type 1 (HIV-1)-infected women in the Women's Interagency HIV Study, a multicenter prospective study. HIV-1 infected women receiving HAART were less likely to have eczema, folliculitis, tinea pedis, and xerosis than were women who had not initiated HAART, independent of CD4+ cell count. Participants who had a prior history of a nadir CD4+ cell count of <200 cells/microL and recent CD4+ cell counts of 200-349 cells/microL were more likely to have eczema and xerosis than were women with a nadir CD4+ cell count of >200 cells/microL and recent CD4+ cell counts of >349 cells/microL. An HIV-1 RNA load of >100,000 copies/mL was associated with increased prevalence of herpes zoster infection (odds ratio, 6.10; 95% confidence interval, 2.00 18.65). History of injection drug use was associated with a higher prevalence of onychomycosis, tinea pedis, and xerosis. Molluscum contagiosum was more prevalent among younger women. PMID- 14765355 TI - Poor validity of self-reported hepatitis B virus infection and vaccination status among young drug users. AB - Self-reported hepatitis B virus (HBV) infection status and immunization status were compared with HBV serological markers among 324 young injection drug users (IDUs) and noninjection drug users (NIDUs). The overall validity of self-reported status was poor; 52% claiming to be vaccinated were actually susceptible to HBV. There was no difference in validity of self-reported HBV status between IDUs and NIDUs. Clinicians should adopt a "Don't Ask, Vaccinate" vaccination policy for young drug users. PMID- 14765356 TI - Serologic response and antibody-titer decay in adults with pertussis. AB - Pertussis is a frequent and significant illness in adults. Because acellular pertussis vaccines for use in adolescents and adults have now been developed, it is important to compare serologic responses in adults after infection with serologic responses in adults after vaccination. We measured IgG and IgA antibodies to 4 Bordetella pertussis antigens at approximately 6-month intervals for 28 months in 11 adults with pertussis. After reaching peak levels, titers of antibody to pertussis toxin decreased more than did titers of antibodies to filamentous hemagglutinin, pertactin, and fimbriae type 1 and type 2. Although studies of adults who have been vaccinated with acellular pertussis vaccines have had shorter follow-up periods than studies of adults with pertussis infection, the antibody decay patterns are similar in both groups. PMID- 14765358 TI - Accurate diagnosis of infection with Histoplasma capsulatum var. duboisii. PMID- 14765359 TI - Hypothyroidism in HIV-infected patients who have or have not received HAART. PMID- 14765360 TI - Arthrobacter woluwensis subacute infective endocarditis: case report and review of the literature. AB - We report a case of endocarditis due to Arthrobacter woluwensis and review the published reports of Arthrobacter species isolated from human clinical samples. A 39-year-old injection drug user presented with fever and a new heart murmur. A. woluwensis was isolated from blood cultures, and a diagnosis of subacute infective endocarditis of the native mitral valve was made. The patient was successfully treated with a 6-week course of intravenous teicoplanin. From our review of the literature, we were able to retrieve data on 41 cases of Arthrobacter species isolated from human clinical samples. However, Arthrobacter species was documented as a cause of human disease on only 5 other occasions (2 cases of bacteremia, 1 case of postoperative endophthalmitis, 1 case of a Whipple disease-like syndrome, and 1 case of phlebitis). Because of the difficulty of identifying Arthrobacter strains by conventional biochemical assays, it is likely that infections with these coryneform bacteria are underreported. PMID- 14765361 TI - Acute renal failure on immune reconstitution in an HIV-positive patient with miliary tuberculosis. AB - Immune reconstitution syndrome following HAART in human immunodeficiency virus (HIV)-infected patients is characterized by inflammatory worsening of organ functions despite improvement in HIV surrogate markers of HIV infection. We describe a patient with miliary tuberculosis and urinary shedding of acid fast bacilli who developed acute renal failure 8 weeks after initiation of antituberculosis therapy and 6 weeks after initiation of HAART. The diagnostic workup and further course of disease implicated immune reconstitution syndrome as the cause of acute renal failure. PMID- 14765362 TI - Aeromonas meningitis complicating medicinal leech therapy. AB - Medicinal leeches have an important and expanding role in medicine, but infection can complicate their use. We describe a unique case of Aeromonas meningitis associated with the use of leech therapy to salvage a skin flap after central nervous system surgery. PMID- 14765366 TI - Intestinal ischemia reperfusion injury and multisystem organ failure. AB - Intestinal ischemia-reperfusion is a common pathway for many diseases in infants, children, and adults, and this may lead to multiple organ dysfunction syndrome and death. While several studies have investigated reperfusion injury in cardiac, cerebral, and hepatic disease, limited work has been published on intestinal ischemia-reperfusion and its multiorgan effects. The authors have developed models of intestinal ischemia-reperfusion in rats and have demonstrated that intestinal reperfusion causes liver energy failure at normothermia. This is followed by 100% mortality within 4 hours of reperfusion. Moderate hypothermia (32 degrees C to 33 degrees C) induced throughout ischemia and reperfusion prevents liver energy failure, intestinal damage, and neutrophil infiltration in the lungs. Moderate hypothermia in this model of intestinal ischemia and reperfusion prevents mortality. Further studies are needed to establish whether therapeutic hypothermia is a useful intervention in the treatment of infants and children with intestinal injuries caused by ischemia and reperfusion. PMID- 14765365 TI - Heparin-binding epidermal growth factor-like growth factor and intestinal ischemia-reperfusion injury. AB - Intestinal ischemia/reperfusion (I/R) injury affects patients of different ages, especially premature babies and the elderly. The outcome after intestinal I/R is often dismal, which may be attributed to loss of the barrier and immune functions of the intestines, as well as development of secondary injury in remote organs. The available treatment for advanced gut ischemia mandates extensive resection, which may cause growth retardation in infants and nutritional problems in the elderly. Throughout the past decade we have been investigating the potential therapeutic role of heparin-binding epidermal growth factor-like factor (HB-EGF) in intestinal I/R. The mitogenic and chemoattractant functions of HB-EGF formed the initial rationale for our investigations. In addition, HB-EGF is a potent antiapoptotic protein that enables cells and tissues exposed to different apoptotic stimuli to survive hypoxic, oxidative, and nutritional stresses. HB-EGF is known to have a vital role in wound healing and postischemic regeneration in different organs. In the current review, we summarize the results of our findings of the beneficial effects of HB-EGF in intestinal I/R, supported by additional evidence from the literature and an explanation of different possible mechanisms of its actions. Collectively, the data strongly suggest a potential therapeutic role for the use of HB-EGF to treat intestinal ischemic diseases such as I/R and necrotizing enterocolitis. PMID- 14765367 TI - Pathogenesis of Hirschsprung's disease and its variants: recent progress. AB - The enteric nervous system (ENS) is a complex network of interconnected neurons within the wall of the intestine that controls intestinal motility, regulates mucosal secretion and blood flow, and also modulates sensation from the gut. The cells that form the ENS in mammals are derived primarily from vagal neural crest cells. During the past decade there has been an explosion of information about genes that control the development of neural crest. Molecular-genetic analysis has identified several genes that have a role in the development of Hirschsprung's disease. The major susceptibility gene is RET, which is also involved in multiple endocrine neoplasia type 2. Recently, genetic studies have provided strong evidence in animal models that intestinal neuronal dysplasia (IND) is a real entity. HOX11L1 knockout mice and endothelin B receptor-deficient rats demonstrated abnormalities of the ENS resembling IND type B in humans. These findings support the concept that IND may be linked to a genetic defect. PMID- 14765368 TI - Extracellular control of pancreatic differentiation. AB - Our understanding of basic mechanisms of differentiation has evolved rapidly in the last two decades. Spurred by advances in molecular biology and other research technologies, these advances have become of heightened importance with the recent advent of the possibility of engineering different types of stem cells into needed cell and tissue sources. As pediatric surgeons, we have the potential to play a key role in interfacing between the basic science necessary to understand differentiation processes, and its application at the bedside. In this brief article, we outline our in-depth analysis of mechanisms of basic differentiation of pancreatic precursor cells in an effort to better understand ways in which we can engineer a stem cell pool to form mature pancreatic cells. PMID- 14765369 TI - Control and regulation of pulmonary hypoplasia associated with congenital diaphragmatic hernia. AB - Control of fetal lung growth and development is exquisitely orchestrated and regulated. Branching morphogenesis is carefully choreographed with cell growth, proliferation, differentiation, and apoptosis in a spatially and temporally dependent manner. Some of the signals and pathways mediating these events have recently been uncovered, but much remains unknown. The precise etiologic derangements that give rise to pulmonary hypoplasia in congenital diaphragmatic hernia remain elusive. Some clues have been discovered in developmental and signaling pathways that include receptor tyrosine kinase growth factors, homeobox genes, transcription factors, airway distension, retinoid signaling, and oxidation-reduction. PMID- 14765370 TI - Gene transfer to the tracheobronchial tree: implications for fetal gene therapy for cystic fibrosis. AB - Gene transfer to the tracheobronchial tree has been an active area of investigation since the discovery of the genetic defect in the cystic fibrosis transmembrane conductance regulator over two decades ago. Cystic fibrosis (CF) is the most common lethal monogeneic disorder for which there is no cure short of lung transplantation. The ultimate goal of gene therapy in CF is to achieve efficient gene transfer at a level and distribution in target tracheobronchial epithelial cells and submucosal gland cells, which results in the genotypic and phenotypic correction of CF. This article reviews the current challenges and limitations of postnatal gene therapy to the tracheobronchial tree, and the potential advantages of fetal gene therapy for CF. We review recent work with novel viral vectors to achieve extremely efficient gene transfer in target cells in the respiratory epithelium and submucosal glands in models that are representative of the developing human fetal trachea. Finally, we will examine the prospects for, limitations of, and regulatory challenges facing the translation of fetal gene therapy from research to clinical application. PMID- 14765371 TI - Antiangiogenic therapy for the treatment of pediatric solid malignancies. AB - Although the past 30 years have seen remarkable progress in the treatment of childhood malignancies, not all types of cancer have enjoyed this improvement in prognosis. Because of this, clinical trials are ongoing in which novel treatment approaches are being evaluated, including immunotherapy, radionuclide therapy, and the use of agents that induce tumor apoptosis or differentiation. Additional treatment strategies are needed, however. One such strategy involves the use of angiogenesis inhibitors. Angiogenesis is the biologic process of new blood vessel formation. In addition to occurring as part of several normal, physiologic processes, angiogenesis is an essential component of a number of pathologic conditions, including cancer. Compelling data suggest that inhibition of angiogenesis can not only prevent tumor-associated neovascularization but also affect tumor growth and spread. An anticancer approach in which the tumor-induced new blood vessels are targeted is particularly appealing for several reasons. First, despite the extreme molecular and phenotypic heterogeneity of human cancer, it is likely that most, if not all, tumor types require neovascularization to achieve their full malignant phenotype. Therefore, antiangiogenic therapy may have broad applicability for the treatment of human cancer, as well as the many other pathologic processes that depend on angiogenesis. Second, the endothelial cells, although rapidly proliferating, are inherently normal with a very low rate of mutation. They are, therefore, unlikely to evolve an angiogenesis inhibitor-insensitive phenotype. This is in distinction to the rapidly proliferating tumor cells that do undergo a high rate of spontaneous mutation and therefore can readily generate drug-resistant clones. This review discusses progress in the development of antiangiogenic therapy for the treatment of pediatric solid tumors. PMID- 14765372 TI - Neuroblastoma and dendritic cell function. AB - Neuroblastoma, the most common extracranial solid tumor of childhood, remains a challenge for clinicians and investigators in pediatric surgical oncology. The absence of effective conventional therapies for most patients with neuroblastoma justifies the application of novel, biology-based, experimental approaches to the treatment of this deadly disease. The observation that some aggressive neuroblastomas, particularly in infants, may spontaneously regress suggested that immune-mediated mechanisms may be important in the biology of this disease. Advances in the understanding of the cognate interactions between T cells, antigen-presenting cells and tumors have demonstrated the sentinel role of dendritic cells (DC), the most potent antigen presenting cells, in initiating the cellular immune response to cancer. Until recently the function of DC in pediatric solid tumors, especially neuroblastoma, had not been extensively studied. This review discusses the role of DC in initiating and coordinating the immune response against cancer, the ability of neuroblastoma to induce DC dysregulation at multiple levels by inhibiting DC maturation and function, and the current vaccine strategies being designed to employ the unique ability of DC to promote neuroblastoma regression. PMID- 14765377 TI - State-of-the-art treatment of high-grade brain tumors. AB - The incidence of primary brain tumors has rapidly increased in recent years. The current standard of care for patients with high-grade malignant glioma is resection followed by radiotherapy. However, the use of adjuvant chemotherapy and the standard of care at first relapse are still under debate for patients with glioblastoma multiforme and anaplastic astrocytoma. Meta-analyses have suggested that adjuvant chemotherapy, specifically with nitrosourea-based regimens, is associated with improved survival. However, no randomized, controlled trial has shown a clear advantage for adjuvant chemotherapy in these patients. Cumulative toxicity associated with both radiotherapy and chemotherapy, as well as resistance to nitrosourea-based regimens related to exposure in the adjuvant setting, prevent the use of radiotherapy and nitrosourea-based regimens at first relapse. The combination of procarbazine, carmustine, and vincristine (PCV) has shown activity at first relapse in patients who have not received adjuvant chemotherapy. Temozolomide (Temodar [US], Temodal [international]; Schering Plough Corporation, Kenilworth, NJ) has shown activity at both first and second relapse in patients who have received prior nitrosourea-based regimens. The better safety profile of temozolomide suggests that it may be preferred to PCV for treatment of patients with recurrent high-grade malignant glioma. Additional randomized, controlled trials are needed to fully define the best option for first-line chemotherapy in both the adjuvant and recurrent settings in patients with high-grade malignant glioma. PMID- 14765378 TI - Factors influencing survival in high-grade gliomas. AB - Multivariate analyses have consistently identified a number of patient, tumor, and treatment variables that are significant prognostic factors for overall survival in patients with high-grade glioma. Age, performance status, mental status, tumor grade and histology, and extent of surgical resection are among the most significant prognostic factors influencing survival. Recently, we showed that normal versus abnormal baseline Folstein mini-mental status examination score is a significant prognostic factor in younger patients (age /=18 months by 15%. Only with large meta-analyses did the addition of chemotherapy achieve a statistically significant improvement in median survival. Currently there is no means of identifying which patients will benefit from adjuvant chemotherapy, but nitrosoureas and temozolomide are well tolerated in most patients, justifying the administration of adjuvant chemotherapy to all newly diagnosed patients with malignant glioma. PMID- 14765381 TI - Prognostic factors for low-grade gliomas. AB - Low-grade gliomas are a heterogenous group of diseases characterized by relatively slow-growing primary brain tumors of astrocytic and/or oligodendroglial origin. Many patients present with easily controlled seizures and remain stable for years, whereas others progress rapidly to higher-grade tumors. Several studies have retrospectively investigated tumor-, patient-, and treatment-related prognostic factors in this patient population. Tumor histology, grade, location, contrast enhancement, and molecular markers have been identified as prognostic factors for survival. Likewise, patient age, performance status, and seizure history are patient-dependent prognostic factors. However, although patients who undergo surgical resection and receive adjuvant radiotherapy tend to have improved survival, treatment-dependent prognostic factors have yet to be definitively identified. Recursive partitioning and multivariant analyses have identified a class of patients with good prognosis. Younger patients with good performance status, non-contrast-enhancing tumors (<5 cm), and tumors of oligodendroglial or mixed-oligoastrocytic subtype have improved survival. The European Organisation for Research and Treatment of Cancer has recently developed a prognostic score based on identified prognostic factors to assist in the management of low-grade gliomas. For patients with a favorable (low-risk) score, treatment with radiotherapy or chemotherapy treatment should be withheld until tumor progression. For patients with a high-risk score, treatment at diagnosis may be indicated. However, other than surgery, the optimal types and sequence of therapies are not yet established. Improvements in defining prognostic factors will assist in low-grade glioma management. PMID- 14765382 TI - Radiotherapy in low-grade gliomas: pros. AB - Although surgery is generally accepted as an important initial treatment in patients with low-grade gliomas, the role of postoperative radiotherapy (RT) is the subject of ongoing controversy. What is the most appropriate timing of RT? Is it best to deliver RT immediately after surgery or to watch and wait until disease progression? Numerous retrospective studies have shown that immediate postoperative RT provides a survival benefit. A recently published prospective study conducted by the European Organisation for Research and Treatment of Cancer has also shown that immediate postoperative RT significantly improved 5-year progression-free survival, but overall survival was not improved. Given the uncertainty about the overall survival benefit of immediate RT and the potential for radiation-induced toxicity, it is important to consider prognostic factors when selecting patients. Subgroups of patients with poor prognostic factors are more likely to benefit from immediate postoperative RT. Available data indicate that a carefully balanced therapeutic approach is needed to optimize treatment outcomes in patients with low-grade gliomas. Patients with more aggressive disease and a high risk of malignant transformation will likely benefit from immediate RT, whereas patients with low-risk disease, particularly oligodendrogliomas, can reasonably be observed. However, one cannot exclude the possibility that delaying RT may result in a worse long-term clinical outcome. PMID- 14765383 TI - Radiotherapy in low-grade gliomas: Cons. AB - Although postoperative radiotherapy (RT) is widely used in patients with low grade gliomas (LGGs), its clinical benefit remains unproven. Because LGGs include diseases with many different histologies, cytogenetic patterns, and natural histories, evaluating the effect of any treatment for LGGs is difficult. Analysis of prognostic subgroups has shown median survival rates ranging from 1 to 10 years. The clinical benefit of postoperative RT in LGGs in retrospective studies is confounded by the heterogeneity of LGGs and treatment-related variables. These studies report conflicting results, most likely because of differences in patient populations. The only prospective, randomized trial comparing postoperative RT with observation failed to show a survival benefit but showed a modest, although statistically significant, improvement in time to progression. Given that two thirds of patients in the observation group received RT at progression, the absence of a survival benefit suggests that RT is active in LGGs, but that delayed RT may provide the same survival advantage as postoperative RT. Moreover, high-dose or whole-brain RT can cause cognitive deficits. Our opinion is that postoperative RT should not be administered routinely to patients with LGGs. If RT is deemed necessary, such as in progressive or inoperable disease causing neurologic symptoms, a total dose of /=60 years of age. Elderly patients represent an important subgroup that accounts for approximately half of primary central nervous system lymphoma cases. Radiotherapy (RT) alone has produced disappointing results in the elderly. Improved clinical outcomes have been achieved with high-dose methotrexate-based chemotherapy regimens in combination with whole-brain RT. Unfortunately, combination therapy exposes elderly patients to a high risk of severe delayed neurotoxicity in up to 80%. Chemotherapy alone may be the best approach for treating elderly patients and appears to be more effective than RT, while considerably reducing the risk of neurotoxicity. Clinical studies are needed to define the optimal chemotherapy regimen for the elderly in an effort to delay or completely avoid the need for RT. PMID- 14765388 TI - The treatment of elderly patients with high-grade gliomas. AB - Elderly patients with cancer are frequently excluded from cancer therapy trials, treated suboptimally, or not treated at all because of the widely held belief that elderly patients do not tolerate chemotherapy and/or radiotherapy (RT) as well as younger patients. Excluding elderly patients from conventional treatment, chemotherapy in particular, is often based on ad hoc decisions rather than on sound scientific data. Malignant gliomas are the most common primary brain tumors in adults, and the age-adjusted incidence of high-grade gliomas has increased over recent years, especially in the elderly. However, few investigators focus on the treatment of high-grade gliomas in the elderly. Data from retrospective studies and meta-analyses suggest that elderly patients with high-grade gliomas have a poorer outcome than younger patients, possibly because of the presence of comorbidity, resistance to cancer therapy, genetic aberrations, different histology, neurodegeneration, or age discrimination. The optimal treatment of elderly patients with high-grade gliomas has not been determined. Surgical debulking and postoperative RT are associated with a significant increase in survival among elderly patients who are in good clinical condition. A recent report has shown that treatment with temozolomide (Temodar [US], Temodal [international]; Schering-Plough Corporation, Kenilworth, NJ) plus RT provides a significant survival benefit compared with RT alone and a significantly improved time to progression compared with RT plus standard chemotherapy (lomustine, procarbazine, and vincristine). Further, temozolomide was well tolerated in the elderly patient population and was less toxic than standard chemotherapy. Therefore, it could be recommended that a full course of RT be followed by adjuvant temozolomide in elderly patients with good prognostic factors. Further, temozolomide alone could be considered as a treatment option for elderly patients with glioblastoma with poorer performance status and for patients who cannot tolerate RT. Results from larger prospective trials will determine the optimal role of chemotherapy, particularly temozolomide, in elderly patients with malignant gliomas. PMID- 14765389 TI - Radiotherapy for malignant gliomas in the elderly. AB - While fractionated external beam radiotherapy remains the principal treatment in patients with histologically verified malignant glioma, its use in older patients and in patients with adverse prognostic features has not been subject to randomized studies. Hypofractionated partial brain irradiation offers a short, well-tolerated treatment with a palliative benefit for patients with a predicted median survival of less than 6 months. To assess its true efficacy in terms of survival and quality-of-life gain requires a formal randomized comparison with supportive care either alone or in combination with palliative chemotherapy and with radical radiotherapy. On present evidence, palliative radiotherapy remains the appropriate treatment for older patients and those with adverse prognostic features, but the final choice should be based on the patient's wishes moderated by the clinician's perception of the gain of treatment. PMID- 14765390 TI - Should radiotherapy be standard therapy for brain tumors in the elderly? Cons. AB - Elderly patients with malignant gliomas present unique treatment challenges. Data are limited on the efficacy of various treatment modalities for high-grade gliomas in the elderly patient population, particularly for patients over 70 years of age, and the most appropriate treatment strategies remain a matter of debate. These patients generally have a poor clinical course and lower tolerance to therapy compared with younger patients. Standard radiotherapy (RT) may provide a survival benefit for patients with a good Karnofsky performance status (>/=70); however, RT is associated with significant neurologic toxicity in elderly patients. Some argue that the apparent modest clinical benefits of RT do not outweigh the risks and inconvenience of the standard schedule. Clearly, more effective and better-tolerated treatments are needed. Chemotherapy agents with good safety profiles, such as oral temozolomide (Temodar [US], Temodal [international]; Schering-Plough Corporation, Kenilworth, NJ), are being considered as alternatives to RT. When used as single-agent therapy, temozolomide induced objective responses and improved neurologic status in elderly patients with newly diagnosed, unresected glioblastoma. For patients with good performance status, treatment options should be discussed and the risks and potential benefits of RT carefully considered. Furthermore, prospective randomized studies should be performed to define the best therapeutic option for efficacy, toxicity, convenience, and quality of life. PMID- 14765391 TI - New frontiers in the treatment of malignant glioma. AB - Despite many technologic advances in neuroimaging, neurosurgery, and radiation therapy, there has been little improvement in survival for patients with malignant glioma. Given the failure of traditional treatment approaches to significantly improve survival in patients with malignant gliomas, research in this field has focused on gaining a better understanding of the molecular pathogenesis of gliomas with the goal of identifying novel drug targets and therapeutic strategies. The influence of molecular genetics on response and survival has been best shown in oligodendrogliomas. The hallmarks of low-grade and anaplastic oligodendrogliomas are their exquisite sensitivity to chemotherapy and favorable prognosis, which are correlated with loss of heterozygosity of chromosomes 1p and 19q. Together, loss of heterozygosity of 1p and 19q appears to confer responsiveness to chemotherapy and to correlate with improved survival. In contrast, deletion of the CDKN2A gene is correlated with poor response to chemotherapy and poor survival, and loss of heterozygosity of chromosome 10q is associated with shorter progression-free and overall survival compared with intact 10q. A variety of other molecular genetic abnormalities have been recognized in gliomas, and these genetic changes are not only important prognostic factors, but also offer new therapeutic targets. With the increased use of improved surgical and radiotherapy techniques and targeted biologic therapy over the next 20 years, many patients with malignant gliomas may be cured or their disease may be controlled for the long term. Molecular profiling of patients using gene chip technology will likely become commonplace, and many patients will receive a tailored treatment regimen based on the unique genetic profile of their tumor. PMID- 14765392 TI - The European Organisation for Research and Treatment of Cancer strategy for new drug development in brain tumors. AB - The European Organisation for Research and Treatment of Cancer (EORTC) has carried out clinical trials for nearly 30 years. During this time, several disease-oriented groups within the EORTC have been shaped by rapidly changing regulations in the area of drug development. Continual networking, dissemination of knowledge, and the establishment of specialized administrative units have allowed the EORTC to play a leading role in the development of improved treatment options for patients with brain tumors. Recently, the EORTC Brain Tumor Group and the EORTC New Drug Development Group have been integrated to create a network of institutions dedicated to screening new chemotherapy agents for the treatment of high-grade gliomas. Indeed, several new regimens and agents have already been investigated by this collaboration. Most notable are the recently closed study on temozolomide (Temodar [US], Temodal [international]; Schering-Plough Corporation, Kenilworth, NJ) in the adjuvant setting and in combination with radiotherapy in patients newly diagnosed with glioblastoma multiforme, and the ongoing study on oral STI571 for the treatment of gliomas. Within the current networks, a well defined strategy has been implemented to select specific and targeted agents for the drug screening process. By sharing their expertise with the EORTC networks, medical oncologists, neuro-oncologists, and radiologists can assist in the clinical development of potentially active agents in patients with brain tumors. PMID- 14765394 TI - Chromoendoscopy with methylene blue and the risk of DNA damage. PMID- 14765395 TI - Failure to replicate the association of the novel NOD2/CARD15 haplotype (S268P JW1) to Crohn's disease in the Jewish Israeli population. PMID- 14765396 TI - The frame-shift mutation of the NOD2/CARD15 gene is significantly increased in ulcerative colitis: an *IG-IBD study. PMID- 14765397 TI - Discussion on spontaneous resolution of chronic hepatitis C virus after withdrawal of immunosuppression. PMID- 14765398 TI - Healthy offspring in parents both receiving thiopurines. PMID- 14765399 TI - Adenomas of the ampulla of vater in familial adenomatous polyposis: Christian Theodore responds. PMID- 14765400 TI - Gastric electrical stimulation for refractory gastroparesis. PMID- 14765401 TI - Complexity and cancer. PMID- 14765402 TI - Diclofenac reduces the incidence of acute pancreatitis after endoscopic retrograde cholangiopancreatography. PMID- 14765403 TI - Hepatitis B virus genotypes and core promoter variant. PMID- 14765404 TI - Accepting the diagnosis. Lia Edwards. PMID- 14765405 TI - In sickness and in health. What's my role in my spouse's diabetes care? PMID- 14765406 TI - American Diabetes Association Resource Guide 2003. New products. PMID- 14765407 TI - American Diabetes Association Resource Guide 2003. Class action: type 2 pills update. PMID- 14765408 TI - American Diabetes Association Resource Guide 2003. Insulin. PMID- 14765409 TI - American Diabetes Association Resource Guide 2003. Insulin delivery. PMID- 14765410 TI - American Diabetes Association Resource Guide 2003. Blood glucose monitors and data management. PMID- 14765411 TI - American Diabetes Association Resource Guide 2003. Products for treating low blood sugar. PMID- 14765412 TI - American Diabetes Association Resource Guide 2003. Urine testing. PMID- 14765413 TI - American Diabetes Association Resource Guide 2003. Carrying cases. PMID- 14765414 TI - American Diabetes Association Resource Guide 2003. Medical identification products. PMID- 14765415 TI - American Diabetes Association Resource Guide 2003. Manufacturers & exclusive distributors. PMID- 14765416 TI - A is for A1C. Are your numbers where they should be? Part 2. PMID- 14765417 TI - Research profile. A new tack. Overcoming low literacy in minorities. PMID- 14765419 TI - Healthy eating. Rice is nice. PMID- 14765418 TI - An apple a day still keeps the doctor away. PMID- 14765420 TI - Body image. Whose body? Whose standards? PMID- 14765421 TI - Fast food & restaurant fare. Hardee's. PMID- 14765422 TI - The diabetes ABC's. B stands for blood pressure. Part 3. PMID- 14765423 TI - Looking for a cure. So far, Ellen Berty's islet transplantation has been a success. PMID- 14765424 TI - The Edmonton Protocol. The future of islet transplantation? PMID- 14765425 TI - Self-care secrets. What ails you? If you have diabetes, some over-the-counter cold and cough medicines can do more harm than good. PMID- 14765426 TI - Self-care secrets. Skin deep. Practices and products to make you comfortable in your own skin. PMID- 14765427 TI - Self-care secrets. Focus on feet. Here's why people with diabetes should be obsessed with their feet. PMID- 14765428 TI - Self-care secrets. Save your smile. Here are great tips for keeping your mouth happy and healthy. PMID- 14765429 TI - Self-care secrets. Giving up smoking for good. Use these strategies to help you stop smoking. PMID- 14765430 TI - Self-care secrets. Multivitamin vitality. Think vitamins are for kids? Think again. PMID- 14765431 TI - Self-care secrets. Fabulous fiber. If you have diabetes, fiber is your friend. PMID- 14765432 TI - Self-care secrets. Talking about urinary incontinence. You're not alone. Loss of bladder control is more common than you think. PMID- 14765433 TI - Self-care secrets. At-home health devices. Here's a look at do-it-yourself devices for managing diabetes and other health conditions. PMID- 14765434 TI - Research profile. Cardiovascular care. Solving the puzzle. PMID- 14765436 TI - Heart rates. Hit your target. PMID- 14765435 TI - Partners in equality. Companies that treat people with diabetes fairly. PMID- 14765437 TI - Vaginitis, yeast infections, & diabetes. PMID- 14765438 TI - Hypertension "essentials". PMID- 14765439 TI - Medicare covers foot exams. PMID- 14765440 TI - Fast food & restaurant fare. Kentucky Fried Chicken. PMID- 14765441 TI - Just for teens. Hypo unawareness. PMID- 14765442 TI - Healthy eating. Main dish salads. PMID- 14765443 TI - When your insurance company denies a claim. PMID- 14765444 TI - Carole Gale into Africa. PMID- 14765446 TI - Women's health exchange. UTIs. Prevention and treatment. PMID- 14765445 TI - Research profile. Best friends and diabetes. PMID- 14765447 TI - What's for dinner? Pass the PPARs please! PMID- 14765448 TI - Healthy eating. Grilling. Fire up your fruits and vegetables. PMID- 14765449 TI - Fast food and restaurant fare. Blimpie. PMID- 14765450 TI - Derek Shockro saving lives. PMID- 14765451 TI - Everybody into the water. PMID- 14765452 TI - Frozen shoulder & diabetes. Frozen shoulder affects 20 percent of people with diabetes. Proper treatment can help you work through it. PMID- 14765453 TI - Simple steps to fitness. Why exercise matters. Exercising does more than burn calories--it can also boost your physical, mental, and even financial well-being. PMID- 14765454 TI - Simple steps to fitness. Adapting exercise to your health. Here are some tips on finding the right exercise for YOU. PMID- 14765455 TI - Simple steps to fitness. Nutrition for exercise. Checking your blood sugar tells you when and if you need to alter your eating habits. PMID- 14765456 TI - Simple steps to fitness. Barriers to exercise (and how to get past them). Everything from time constraints to safety concerns can discourage you from exercising. Don't let them! PMID- 14765457 TI - Simple steps to fitness. Your walking plan. No matter what shape you're in, here's a plan to get you moving. PMID- 14765458 TI - Simple steps to fitness. Secrets of strength. Strength training is as important to weight loss and fitness as aerobic exercise is, but many people ignore it. Here's how to get started. PMID- 14765459 TI - Research profile. Cholesterol. The eye-kidney connection. PMID- 14765460 TI - Healthy eating. Keep it simple. Eating the Mediterranean way. PMID- 14765461 TI - From heart to sole. Diabetes and the heart-foot connection. PMID- 14765462 TI - Bringing up baby. Pregnancy and diabetes. PMID- 14765464 TI - Losing weight. PMID- 14765463 TI - PAD. The "vascular" in cardiovascular disease. PMID- 14765465 TI - Low-carb diets. The debate continues. PMID- 14765466 TI - Living longer. Six steps for adding years to your life. PMID- 14765468 TI - Fast food & restaurant fare. Long John Silver's. PMID- 14765467 TI - Eating disorders. A meter tells the real story. PMID- 14765469 TI - A father first. Colorado Rockie third baseman never gave much thought to diabetes. Until Hannah. PMID- 14765471 TI - Color me healthy. Put a rainbow on your plate. PMID- 14765470 TI - When your child is diagnosed. Your child's diabetes can cause a tidal wave of emotions. Here's how to stay on an even keel. PMID- 14765473 TI - Foot ulcers. New wound gel may aid healing. PMID- 14765472 TI - Research profile. Obesity in toddlers. Genes and chemistry, not lifestyle? PMID- 14765474 TI - HIV vaccines: current challenges and future directions. AB - Volume seven of the Review will mark the tenth anniversary of the Canadian HIV/AIDS Legal Network with a series of articles that describe past developments and future directions in several areas of policy and law related to HIV/AIDS. The following article is the first of these, discussing current challenges and future directions in the development of and access to HIV vaccines. It argues that governments are under public health, ethical, and legal obligations to develop and provide access to HIV vaccines. It further explains what is required for governments to fulfill their obligations: additional commitment and resources for HIV vaccine development in the context of increased global research and development regarding diseases of the poor; increased support and advocacy for partnerships to develop HIV vaccines; enhanced regulatory capacity in every country to review, approve, and monitor HIV vaccines; and assurance of global supply of, procurement of, delivery of, and access to vaccines in the context of efforts to increase global access to public health measures and technologies. PMID- 14765475 TI - Drug policy in Canada--the way forward. AB - This article is one of a series commissioned to mark the tenth anniversary of the Canadian HIV/AIDS Legal Network, discussing past developments and future directions in areas of policy and law related to HIV/AIDS. It takes a critical look at Canada's drug policy. Despite calls for a balanced approach focused on reducing drug-related harm, Canada's method of dealing with problems of illicit drug use has remained prohibitionist in nature, and by far the greatest part of federal funding is devoted to supply-reduction initiatives. Considerable changes in policy and law are needed to significantly reduce the harms associated with injection drug use in Canada. These include developing a comprehensive and integrated strategy, exploring alternative legal frameworks, piloting innovative approaches to reducing injection-related harms, and investing in broad social policies that address the determinants of injection drug use. PMID- 14765476 TI - Anniversaries--but what have we learned? PMID- 14765477 TI - HIV-related stigma and discrimination--the epidemic continues. AB - This article is one of a series commissioned to mark the tenth anniversary of the Canadian HIV/AIDS Legal Network, discussing past developments and future directions in areas of policy and law related to HIV/AIDS. It looks at HIV related stigma and discrimination. The article summarizes the present situation as described in reports from numerous countries throughout the world. It reviews the institutional, non-institutional, and structural dimensions of HIV-related discrimination. It also identifies some essential components of anti discrimination efforts: legal protection; public, workplace, and health-care programs; community mobilization; and strategizing on the determinants of health. PMID- 14765479 TI - Canada: Privacy Commissioner finds "egregious" violation of inmates' confidentiality. PMID- 14765478 TI - Canada: federal prison system expands access to methadone. PMID- 14765480 TI - Russia: prisoners sue because of publication of medical information. PMID- 14765481 TI - Canada: innovative educational tools on hepatitis C and prisons: the SHARP approach. PMID- 14765482 TI - Canada: recent studies confirm prevalence of injection drug use in prisons. PMID- 14765483 TI - HIV vaccines in Canada: legal and ethical issues--an overview. AB - In July 2002 the Legal Network released an overview paper on legal and ethical issues related to an HIV vaccine in Canada. The paper, which is based on a more detailed report prepared in collaboration with the Centre for Bioethics of the Clinical Research Institute of Montreal, calls for the establishment of a Canadian HIV Vaccine Plan. PMID- 14765484 TI - New immigration law comes into effect as regulations finalized. AB - The new regulations make it easier for same-sex partners to immigrate to Canada. However, the government has rejected suggestions to change the way it determines whether potential immigrants living with HIV/AIDS or other health conditions would place excessive demands on Canada's health and social services. PMID- 14765485 TI - Government delays release of medical marijuana supply. AB - The federal government's initiative to make marijuana available for medical use continues to run into problems and delays. In a recent development, the first crop produced by the government's designated grower turned out to be too impure to use. The delays have led to the launch of a lawsuit against the federal government. PMID- 14765486 TI - Report recommends that HIV become reportable in BC. AB - The Provincial Health Officer has recommended that HIV be nominally reportable and that a well-funded partner notification system be established. However, he also recommended that people be allowed to choose non-nominal HIV testing. PMID- 14765487 TI - Female injection drug users in Vancouver face higher risks of HIV infection. PMID- 14765488 TI - Business group drops opposition to drug users' health centre. AB - A legal intervention by the Vancouver Area Network of Drug Users has forced the Community Alliance to abandon a petition against a health contact centre in Vancouver's Downtown Eastside. PMID- 14765489 TI - Justice reduced: legal aid slashed in BC. AB - Serious cuts to legal aid services in British Columbia will have a devastating impact on people with HIV/AIDS and the organizations that serve them. PMID- 14765491 TI - Inaccurate results lead to removal of rapid HIV tests. PMID- 14765490 TI - Ontario set to introduce new privacy bill. AB - The Ontario government's proposed new legislation is an improvement over previous attempts, but there are still problems. PMID- 14765492 TI - Canada establishes Global Health Research Initiative. PMID- 14765493 TI - BC introduces changes to disability assistance. PMID- 14765494 TI - Ontario commission criticizes insurance industry record on human rights. PMID- 14765495 TI - HIV/AIDS organizations make representations to Romanow Commission. PMID- 14765496 TI - United Nations Expert Panel rejects peacekeeper HIV testing. PMID- 14765497 TI - Namibian Parliament overrides SADC panel, labour court on military testing. PMID- 14765498 TI - Expert meeting on HIV vaccine research, development, and access. PMID- 14765499 TI - AAVP holds ethics, law, and human rights symposium in Durban. PMID- 14765500 TI - Caribbean countries address legal, ethical, human rights issues. PMID- 14765501 TI - SADC parliamentarians discuss HIV/AIDS, legislation, and human rights. PMID- 14765503 TI - South Africa HIV/AIDS training for Bench's "ideological virgins". PMID- 14765502 TI - CIDA funds Kenya, Zambia AIDS law groups. PMID- 14765504 TI - Ukrainian law criminalizes sex work. PMID- 14765505 TI - Ireland: compensation deal regarding HIV-tainted blood products. PMID- 14765506 TI - Ontario court recognizes constitutional equality rights of people "in receipt of social assistance". AB - On 13 May 2002, the Ontario Court of Appeal released its unanimous decision in the Falkiner case, which declared the definition of "spouse" in Ontario's social assistance legislation to be unconstitutional because it discriminates on the basis of sex, marital status, and receipt of social assistance. The case is significant for people living with HIV/AIDS, given that a high proportion live on social assistance and thus face discrimination. PMID- 14765507 TI - Blood or blood products: Minister's decision to deny application to Extraordinary Assistance Plan upheld. AB - In April 2002, the Federal Court of Canada decided the federal Minister of Health does not have the authority to award compensation under the Extraordinary Assistance Plan unless an independent medical review board finds that the applicant was infected with HIV as a result of the receipt of blood or blood products in Canada between 1978 and 1989. PMID- 14765508 TI - Criminal law and HIV transmission/exposure: one new case. One-year sentence for spitting. AB - In a regular column, we have reviewed new developments in the area of criminal prosecutions for HIV transmission or exposure. Since the last issue of the Review, one new Canadian case has come to our attention. PMID- 14765509 TI - HIV-positive child made ward of court after father refuses treatment with antiretroviral drugs. AB - In 1999, we reported a number of cases in Canada, the US, and the UK in which child-welfare authorities have disputed parents' decisions to refuse HIV testing or treatment for their children. This article reports on new developments in one of those cases. On 10 May 2002, the English High Court heard submissions regarding the future of a three- year-old HIV-positive girl who was made a ward of the court after her father refused to allow her to be treated with antiretroviral drugs. PMID- 14765510 TI - English court upholds ban on newspaper publishing identity of HIV-positive health care worker. AB - In the last issue of the Review, we reported about an HIV-positive health-care worker in the UK who brought legal proceedings to prevent the National Health Service from notifying his previous patients that he is infected. In February 2002, the Court of Appeal for England and Wales (Civil Division) upheld a ban on publishing his name and that of the health authority for which he worked. PMID- 14765511 TI - UK court rules government erred in denying housing assistance to woman with HIV. AB - In March 2002, the English High Court of Justice ruled that municipalities have a duty to use their powers to provide assistance where refusal to do so would infringe an applicant's rights under the European Convention on Human Rights and Fundamental Freedoms. PMID- 14765512 TI - El Salvador: activists challenge law allowing pre-employment HIV testing. AB - On 24 October 2001, El Salvador's national legislature passed a law allowing employers to impose pre-employment HIV testing on job applicants, although it also prohibits employers from discriminating against those who test HIV-positive. PMID- 14765513 TI - Indian NGO challenges penal code prohibition of "unnatural offences". AB - On 7 December 2001, the Naz Foundation (India) Trust (NFIT), a non-governmental organization based in New Delhi, filed a petition in the Delhi High Court to repeal the "unnatural offences" section of the Indian Penal Code that criminalizes men who have sex with men. PMID- 14765514 TI - Global access to treatment: achievements and challenges. AB - As mentioned in the previous issue, this section of the Review addresses issues related to improving access to adequate and affordable care, treatment, and support everywhere. It replaces the section previously called "Patents and Prices." In this issue, we feature a review of achievements and challenges in recent years in opening global access to HIV/AIDS treatments. The article - one of a series commissioned to mark the tenth anniversary of the Canadian HIV/AIDS Legal Network, discussing past developments and future directions in areas of policy and law related to HIV/AIDS - describes the developments that recast the debate about access to treatment from one focused on patent entitlements to one focused on the right to health and treatment. It analyzes the role of national and international activism, strategically constructed alliances, and principled leadership in achieving this change. And it discusses continuing obstacles to equitable access to HIV/AIDS treatments for the world's population. PMID- 14765515 TI - First in fear and dread: Cancer control in Saskatchewan, the first decade - 1929 1939. AB - In 1931 Saskatchewan became the first jurisdiction in Canada to introduce a province-wide program for the control of cancer. The program was conceived in 1929 by the cancer committee of the Saskatchewan Medical Association. The provincial government promptly took up the committee's recommendations for centralization of cancer care in two clinics and emphasis on radiotherapy as a mode of treatment. The following year the Saskatchewan Assembly approved The Saskatchewan Cancer Commission Act, as prepared by Health Minister F.D. Munroe, himself a physician. The Regina Cancer Clinic opened at the end of 1931 and the Saskatoon Cancer Clinic at the beginning of 1932. This initial phase of the conception and introduction of cancer control in Saskatchewan is notable for the harmony of agreement among its planners and for the rapidity with which it was introduced. Progress during the remainder of the first decade was much slower, as competing interests and priorities among a larger group of individuals and institutions, together with the Depression, help back the initial momentum. Initially, however, weaknesses accompanied the strengths of the program. Weaknesses included the reluctance of some physicians to cooperate wholeheartedly as they feared the perceived effects of the incursion of a government agency in patient care plus the increasing financial burden placed on the hospitals. The strengths of the program included the existence of the program where none had been present earlier, the availability of a diagnostic and therapeutic service for patients, and a comprehensive system of patient records. In particular, the cancer control program in its first decade was the basis for notable progress in later decades and a model for other jurisdictions. PMID- 14765516 TI - Editorial: An update on the SCAR TRIP Initiative. PMID- 14765517 TI - Live discussion. From here to there: AbetaPP as an axonal transport receptor--how could this explain neurodegeneration in AD. 15 July 2002. PMID- 14765518 TI - Live discussion: Amyloid-beta degradation: the forgotten half of Alzheimer's disease. 12 September 2002. PMID- 14765519 TI - Pitfalls in the measurement of some intermediary metabolites: stabilization of lactate and pyruvate. PMID- 14765520 TI - [The history of the Upper Hess society of natural history and medicine]. PMID- 14765521 TI - [A biography of Christian Gottfried Nees von Esenbeck]. PMID- 14765522 TI - [The prescription as literature]. AB - In the medical writings of antiquity the recipe, a basic form of scientific writing, was considered a literary genre with conventions of its own. In texttype linguistics, however, the recipe plays only a minor role; in studies on the history of literary short texts it is hardly ever mentioned and, furthermore, it has come to the conclusion that the recipe's function has lain solely in the curing of the sick. The present study focuses on its multifunctional character. A collection of widely scattered and hardly accessible recipes shows that the recipe adopted functions beyond giving technical directives. Its multifunctionality in the German literature of early modern times is manifest in medical and cooking jocular recipes, cooking recipes in minnesang, mock recipes aimed at physicians and astrologers, recipes of spiritual-religious content, recipes containing art and literary criticism as well as recipes in a moral satirical or a political-agitative vein. PMID- 14765523 TI - [German colonial plans for Africa in the Nazi era]. AB - The German colonial plans concerning Africa in the era of National Socialism ascribed a central role the sciences. Scientists of all possible fields launched into activities. Especially subjects which were directly related to the practice of colonial policies, e.g. African languages, ethnology, law, economic sciences, and medicine, were developed. There were colonial ambitions at nearly every German university, but there was one which designated to become the centre of colonial sciences: the university of Hamburg. It has to be realized that working in this field of studies protected scientists from being drafted by the army for a long time. PMID- 14765524 TI - [My insanity in the year 1783]. AB - The aim of this paper is twofold: to defend the validity of the manic-depressive psychosis as a disease entity and to make an incursion into the history of psychiatry. The two intentions meet as far as the recognition of a clinical case straight through space and time supports the ontological status of a disease entity. An autobiographical work from 1801 "My insanity in the year 1783" written by the Danish vicar C.S. Andresen, who lived from 1760 to 1832 is briefly summed up. Andresen tells that he as a young student of divinity made a journey from the small town Rorvig in Zeeland to his then place in Copenhagen. The weather was intolerably hot, the coach was uncomfortable, the landscape deserted and empty and just in the hardships the most unhappy and deplorable event happened: He faced the gruesome truth that he was deprived of the most precious gift of his Creator, his reason and faculties. A fellow passenger brought him to an inn where he literally ran up the walls alternately in high spirits and in deep dispair. When he arrived in Copenhagen he was blead, probably by a surgeon, but later that day admitted to King Frederik's Hospital where he was treated with bleds and leechs and baths. However, as the illness dragged on he was according to his own will accompanied by his custodian to his parental home in the Isle of Bornholm where he recovered while nursed by his mother and mother's mother who themselves had suffered from periodical melancholy when they were young. He completed his academic studies and became a highly respected scholar and vicar in the isle of Funen - but almost twenty years after his recovery he was made to suffer the humiliation that he - a true representative of the Age of Enlightment and rationalistic theologician - was suspected to have invoted his insanity by a sinful life in his youth. Thus his work may be regarded as a defensive pamphlet written at a time when a neutral and clinical view of mental diseases had given place to a moralistic and religious concept. The case serves as a modification of the general assumption that psychotic patients in "the classic age of confinement" under all circumstances were brutally treated and locked up. Even the fluent and subjective style is charming and reminds the Danes of Johannes Ewald (1743-81) and perhaps - were it to be translated - the English of Lawrence Sterne (1713-68). Maybe the booklet of just 75 pages is worthy of a translation? PMID- 14765525 TI - [Patient record from the Copenhagen General Hospital in 1855]. AB - Hospital monographs frequently describe the buildings and the leading staff but rarely the patients. However, the 1980s and 1990s called attention to the importance of the perspective of patients of the past. The chapter reports an examination of the quality and extent of information retrieved through a systematic search of patient records from the Copenhagen General Hospital, extending back to 1855. The charts illuminated the social conditions of the patients in addition to their illnesses and the treatments characteristic of the day. Analyzing the state of medical practice revealed in the records, the author tested whether hospitals in reality were no more than gateways to the afterlife. Comparisons among hospitals showed the extent to which treatments varied across institutions. Correlations between the evidence of the patient charts and the ideals of the professional literature at the time revealed the differences between theory and practice. The records yielded important insights into the daily lives of patients admitted 150 years ago. They generally were younger and of lower social class. For reasons of poor hygiene, poor housing, and poor nutrition, they fell victim to the classical diseases of the crowded urban environment, including lung, skin and venereal diseases. Of these, tuberculosis killed most patients admitted to hospital. PMID- 14765526 TI - [Neurosurgery in a small Danish county Hospital 1922-50]. AB - Jacob Nordentoft (1881-1954), head of a remote small county hospital in Denmark 1922-50 (88 beds) was a very skilled surgeon strongly in favour of master class education. He did not at all like subspecialisation in surgery, but admitted that neurosurgery might be an exception. However, he found the neurosurgical technique pretty simple. Meticulous analysis of all his neurosurgical procedures, including spinal cord tumors, cord compression, brain tumors, and myelomeningoceles in the preserved medical records from his time reveal results just as good and often better than in much more specialized clinics. PMID- 14765527 TI - [An hour with Venus-a life with Mercur]. AB - In 1857 the police doctor in Copenhagen received an anonymous letter from a gentleman. He wants the physician to write a popular book about syphilis and describe how men, who were infected could treat themselves. Regarding to the letter this paper examines how the authorities and the danish physicians tried to stop the disease in the 1850's and to see how physicians were arguing for the regular prostitution in the city. The disease was spreadening very rapidly in these years and the physicians were very concerned about the new growing up population -the children, who also were suffering from the disease. No proper treatment was available in the 1850's. The severe cases were still treated with mercury like in the Middle Ages. PMID- 14765528 TI - [Analysis of iris: history and future]. AB - Analysis of the iris has ancient roots, but iridology in its present sense was established by Philippi Meyers in 1670. The discipline was further developed by Ignaz Peczely in 1881 and by Nils Liljequist in 1890. Other studies have been published in Germany, Sweden, Norway, England, France, USA, Australia and elsewhere, and iridology is practiced worldwide. The techniques used are described and the sources of error are mentioned when several photos from the same patient are compared (e.g. the influences of differences in pupil size, magnification and the effect of oblique light). The iris key (irigraphy) and the interpretation of different sights are discussed in some detail; there is little agreement among iridologists. Some iris diagnoses are of little interest to day and some modern diseases are missing in the diagnostic system. Controlled studies reveal that iridology is of no use whatsoever for the detection of cancer and other diseases in the stomach, intestines, kidney, lungs and heart. The reason for the continued popularity of iridology is discussed, and it is concluded that this type of alternative medicine is not harmless. PMID- 14765529 TI - [From the glorious time of homeopathy medicine chests in Nord-Schleswing (South Justland) about 1900]. AB - The article deals with the origin, background and development of homoeopathy over a hundred years span, beginning with its "invention" in the early 19th century (Hahnemann). Leipzig became a center for a huge production of homoeopathic remedies through Dr. W. Schwabe's "Central-Apoteke", founded in 1878, with customers all over the world. The preparation of around one thousand products were described in 1880 in Schwabes "Pharmacopoea homoeopatica polyglotta", in five languages, and this article gives a survey of the principles in the production of the final remedies which were distributed as Schwabes homoeopatic house-chests of many varieties, containing from a few up to more than hundred bottles. Detailed instructions and guidance in the applications of the content of these popular and wide-spread chests were available for laymen from hand-books. However their use vanished gradually in the third decade of the 20th century and later, and the chests were either destroyed or put away. This was also true for the situation in South Jutland (before 1920 a german province called Nord Schleswig), nevertheless the author's research succeeded in detecting thirteen homoeopathic house-chests from the region, purchased about year 1900. An attempt is made to throw light on their origin, their use and their users at a time, when homoeopathy played a role for layman in the treatment of diseases, outside the authorized health service - or as a supplement to this. PMID- 14765530 TI - [German qualified doctors license to continue practice in south Jutland after the reunion. (Application of danish jus practicandi, process and result)]. AB - At the reunion of the southern part of Jutland with Denmark 1920 danish legislation had to be introduced in the new incorporated region. According to a special law doctors educated in Germany could obtain danish medical authorization, if they were born or established in the region before 1.1.1918, under certain circumstances it was possible to dispense from this date. The law meant that all doctors, in the region had to ask for danish authorization. Matrimonial relationship and personal connections were strong arguments for a dispensation, but not always sufficient. In more cases a refusal was given because of a german attitude at the applicant in spite of matrimonial relationship, with the result that the applicant had to leave an established practice and Denmark. 53 doctors were given danish authorization immediately. 8 got an authorization by dispensation and 10 got a refusal. A survey of the applications and the corrected archives shows, that the cases mostly were handled without problems. But the cases, where dispensation and especially refusal were given, were often very complicated and followed by heavy local national reactions. This paper describes the background of the special law and the course of the cases with particular weight on the cases where dispensation or refusal were given. PMID- 14765531 TI - [Athlete or brilliant Professor. Portrait busts in the hall of Rigshospitalet's auditorium]. AB - Nine portrait busts, eight in bronze and one in euville-limestone are placed in the hall of Rigshospitalet's auditorium of former professors from periods of nearly 100 years. The professors were all outstanding personalities who have left their stamp on a whole generation of medical students and doctors. The bust is a fine way, three-dimension and here in normal size or even over-normal size to give a fine character sketch of the professor's qualities. Different and skillful well-known Danish artists had made the busts all in high calibre, showing the art at different periods. PMID- 14765532 TI - [The case that did not serve to enhance the reputation of the doctors involved]. AB - A complicated and long action for damages is outlined after the train accident in Bramming, Western Jutland, July 1913, where 15 passengers died and 16 passengers were seriously injured. The case came to last for two years, especially due to one doctor's exorbitant fee charged for the treatment of three foreign patients. By this, the doctor came in conflict with his colleagues and the local medical association and filed a suit against the state, which he lost. The case exposed an avaricious doctor and weak collegiate opposition. PMID- 14765533 TI - [Something about the history and future use of casuistry]. AB - Already Aristotle was aware of that an act ethically could not be judged alone by its effect, but circumstances should also be considered. This point of view was later taken over by Christianity in the form of the so called casuistry. The use of circumstances in the judgment of an act as regards the moral permissibility was, however, to some extent misused, especially by the Jesuites in the 16th and the first half of the 17th century. In defence of the "Jansenites" Blaise Pascal launched a direct attack on the Jesuites' use of casuistry in spiritual advice of the sinners. His elegant and vitty satire destroyed any talk of using casuistry for a long time, at least in the protestantic Northern Europe. Casuistry was regarded as synonym with sophistry. In 1988 Jonsen and Toulmin has argued that it is only the misuse of casuistry that is to blame. Used in a proper way it still has a future resolving moral problems especially in medical ethics. The present author is of the opinion that some degree of casuistical reasonning unconsciously is used in the present ethical debate, but more awareness hereof may be an advantage in the debate. PMID- 14765534 TI - Stroke outcome and neuroimaging of intracranial atherosclerosis (SONIA): design of a prospective, multicenter trial of diagnostic tests. AB - BACKGROUND AND RELEVANCE: Intracranial atherosclerosis is responsible for 70,000 ischemic strokes each year in the USA. Noninvasive testing such as transcranial Doppler ultrasound (TCD) and magnetic resonance angiography (MRA) to identify intracranial atherosclerosis is in widespread use, but has not been rigorously validated against the gold standard, catheter angiography. The recently NIH funded Warfarin-Aspirin Symptomatic Intracranial Disease (WASID) trial will compare warfarin with aspirin for stroke prevention in patients with intracranial atherosclerosis. WASID requires performance of angiography along with TCD and MRA, providing an opportunity to critically evaluate these noninvasive tests. MAIN OBJECTIVE: The purpose of the Stroke Outcomes and Neuroimaging of Intracranial Atherosclerosis (SONIA) study is to develop the noninvasive diagnosis of intracranial atherosclerosis. The primary aim of SONIA is to define velocity values on TCD and anatomic abnormalities on MRA that identify severe (50 99%) intracranial stenosis of large, proximal arteries seen on catheter angiography. SONIA will define the criteria, or 'cutpoints', for an abnormal TCD or MRA and show that they perform with a reliable positive predictive value (PPV). STUDY DESIGN: SONIA will be conducted in collaboration with WASID. Study wide cutpoints defining positive TCD and MRA have been developed and reviewed by the site investigators of WASID. Hard copy angiography, TCD and MRA generated in WASID will be centrally read in SONIA. TCD and MRA cutpoints seek to achieve a target PPV of 80% for the identification of severe intracranial stenosis on angiography. CONCLUSIONS: Central readings will be used to validate the cutpoints and to develop measures of negative predictive value, and inter- and intra observer variability. Sensitivity and specificity will be determined after adjustment for verification bias and employed in receiver-operator characteristic analyses. SONIA will use these techniques to develop TCD and MRA cutpoints that minimize the clinical consequences of test errors occurring in the noninvasive evaluation of patients with suspected intracranial atherosclerosis. PMID- 14765535 TI - Coenzyme-Q(10) concentration in human phenylketonuria. PMID- 14765536 TI - Molecular genetic analysis of glycogen storage disease type Ia in 26 Chinese patients. AB - Sequence analysis of 26 patients from Mainland China with glycogen storage disease type Ia revealed a high frequency of two mutations in the glucose-6 phosphatase gene. These mutations, 727G>T and R83H, were also found to be in linkage disequilibrium with a polymorphism at position 1176. These findings have implications for carrier detection and prenatal diagnosis of this disease in the Chinese population. PMID- 14765538 TI - Upsetting the balance: lipoprotein receptors in the CNS. PMID- 14765537 TI - Clinical heterogeneity in patients with mutations in the NDUFS4 gene of mitochondrial complex I. AB - A comparison of the clinical presentation, disease course and results of laboratory and imaging studies of all patients so far published with a NDUFS4 mutation are presented. This reveals marked clinical heterogeneity, even in patients with the same genotype. PMID- 14765539 TI - Making sense of gene therapy. PMID- 14765540 TI - Ethics board review of biomedical processes: improving the process. PMID- 14765541 TI - Uncertainties regarding dengue modeling in Rio de Janeiro, Brazil. AB - Dengue fever is currently the most important arthropod-borne viral disease in Brazil. Mathematical modeling of disease dynamics is a very useful tool for the evaluation of control measures. To be used in decision-making, however, a mathematical model must be carefully parameterized and validated with epidemiological and entomological data. In this work, we developed a simple dengue model to answer three questions: (i) which parameters are worth pursuing in the field in order to develop a dengue transmission model for Brazilian cities; (ii) how vector density spatial heterogeneity influences control efforts; (iii) with a degree of uncertainty, what is the invasion potential of dengue virus type 4 (DEN-4) in Rio de Janeiro city. Our model consists of an expression for the basic reproductive number (R0) that incorporates vector density spatial heterogeneity. To deal with the uncertainty regarding parameter values, we parameterized the model using a priori probability density functions covering a range of plausible values for each parameter. Using the Latin Hypercube Sampling procedure, values for the parameters were generated. We conclude that, even in the presence of vector spatial heterogeneity, the two most important entomological parameters to be estimated in the field are the mortality rate and the extrinsic incubation period. The spatial heterogeneity of the vector population increases the risk of epidemics and makes the control strategies more complex. At last, we conclude that Rio de Janeiro is at risk of a DEN-4 invasion. Finally, we stress the point that epidemiologists, mathematicians, and entomologists need to interact more to find better approaches to the measuring and interpretation of the transmission dynamics of arthropod-borne diseases. PMID- 14765542 TI - Understanding chemoinformatics: a unifying approach. PMID- 14765543 TI - Positioning ADMET in silico tools in drug discovery. PMID- 14765544 TI - Identification of a common mutation (Gly194Cys) in both Arab Moslem and Ashkenazi Jewish patients with dihydrolipoamide dehydrogenase (E3) deficiency: possible beneficial effect of vitamin therapy. AB - Dihydrolipoamide dehydrogenase (E3) deficiency with a clinical phenotype and genotype (Gly194Cys homozygous) previously identified only in Ashkenazi Jewish patients, was diagnosed in two Palestinian Arab siblings and two unrelated Ashkenazi Jewish patients. While three of the four patients died in childhood without specific treatment, the surviving patient at age 18 years may have benefited from long-term daily supplementation with a cocktail of riboflavin, biotin, coenzyme Q and carnitine. PMID- 14765545 TI - Two novel mutations in the gene for human alpha-mannosidase that cause alpha mannosidosis. AB - Mutation analysis performed on two Italian patients with alpha-mannosidosis allowed the identification of two new mutations, IVS20-2A>G and 322-323insA. The patients were both homozygous for these mutations. The first mutation causes skipping of exon 21, whereas the second causes a frameshift introducing a stop codon at position 160 of the amino acid sequence. PMID- 14765546 TI - LC-MS-MS analysis of 2-pyridylacetic acid, a major metabolite of betahistine: application to a pharmacokinetic study in healthy volunteers. AB - 1. A sensitive liquid chromatographic-tandem mas spectrometric assay was developed and validated to determine the major metabolite of betahistine, 2 pyridylacetic acid, in human plasma. 2. The analyte was extracted from plasma samples by liquid-liquid extraction and analysed using liquid chromatography tandem mass spectrometry with an electrospray ionization interface. The method has a lower limit of quantitation of 1 ng ml(-1) fir a 0.5-ml plasma aliquot. The intra- and interday precision (relative standard deviation), calculated from quality control (QC) samples, was less than 10%. Accuracy as determined from QC samples was within +/-7%. 3. The validated method was successfully applied to a pharmacokinetic study of betahistine in healthy volunteers. After oral administration of a single dose of 24 mg betahistine mesylate to 20 healthy Chinese male volunteers, Cmax was 339.4 ng ml(-1) (range 77.3-776.4 ng ml(-1)). The t(1/2) was 5.2 h (range 2.0(-1)-11.4h). The AUC(0-t) obtained was 1153.5 ng ml(-1) h (range 278.5-3150.8 ng ml(-1)). The disposition of the metabolite exhibited a marked interindividual variation. 4. The plasma concentrations of the parent drug were less than 0.5 ng ml(-1), suggesting that it undergoes almost complete first-pass metabolism. The reported two active metabolites were not detected in the plasma of any volunteer. Although there is no evidence that the major metabolite has pharmacological activity, the clinical importance of 2 pyridylacetic acid in humans should be reinvestigated. PMID- 14765547 TI - Hyponatremia in older women. PMID- 14765549 TI - Positive pressure breathing by total body negative pressure: a new simulation model for fluid balance in weightlessness? AB - A systems study is proposed to explore correctly the effects of the combination of HDT (Head Down Tilt) and TBNP (Total Body Negative Pressure). PMID- 14765548 TI - Social support and depressed mood in isolated and confined environments. AB - The influence of isolation and confinement on social support and depressed mood was examined in a study of 235 men and women who spent a year at McMurdo Station in Antarctica, and a study of 77 men and women who spent a year at the Amundson Scott South Pole Station. Although availability of support remained unchanged, there was a significant decrease in reported satisfaction with support obtained, as well as a significant increase in depressed mood. Satisfaction with support was inversely associated with depressed mood at the beginning and end of isolation and confinement. At the end of winter, this association varied by source of support. High levels of tension-anxiety, depression and anger preceded an increase in advice seeking, but high levels of advice seeking also preceded an increase in tension-anxiety and depression. Results suggest a significant erosion of social support under conditions of prolonged isolation and confinement, leading to an increase in depressed mood. PMID- 14765550 TI - [Boris Rajewsky (1893-1974). A look back on his life and work]. PMID- 14765551 TI - [Wegener's granulomatosis (morbus Wegener)]. PMID- 14765552 TI - Clinical policy for the initial approach to patients presenting with altered mental status. PMID- 14765554 TI - Sterile filtration: process optimization through real compliance. PMID- 14765555 TI - The influence of incubation conditions in sterility tests. AB - Studies were conducted to evaluate the effects of media, incubation temperature and duration on the detection of bacteria and fungal growth using British, United States and Brazilian compendial sterility test methods. 5 to 50 CFU of nine different microorganisms (including both anaerobic and aerobic bacteria and molds) were used to contaminate test units containing various growth media (sobean-casein digest, thioglycollate, Sabourand and Causen broths). Test units were incubated at temperatures ranging from 12 to 42 degrees C for 1 to 28 days. Inoculations were conducted according to compendial procedures. Optimal detection conditions were obtained at 22 to 32 degrees C over 14 days using soybean casein digest and thioglycollate broths. PMID- 14765556 TI - Role of the wireless thermologger system in the management of the autoclave unequipped with sensor ports. AB - Recently, the wireless thermologger system was developed. It consists of a wireless logger and the attached computer system. The wireless logger has a wireless structure and can be placed anywhere in the sterilizer chamber for thermometry. In the present study, thermal evaluations were performed twice using the wireless thermologger system in the sterilizer chamber of the prevacuum autoclave unequipped with sensor ports when it failed to pass the Bowie-Dick test and underwent mechanical repairs. Thermometry was performed when the Bowie-Dick test cycle was operated. The heat up time was measured in the range of 120.0 134.0 degrees C. The F0 value was calculated with the reference temperature at 121.0 degrees C and Z value at 10 degrees C when the sterilization temperature was over 120.0 degrees C. The first thermal evaluation was performed after three air-leaking points were repaired along with the replacement of the vacuum pump and the thermosensor. It revealed that the heat-up time was 4 min and 42 s and the F0 value was 137.5. After the temperature control systems were adjusted using the process calibrator in the prevacuum autoclave, the second thermal evaluation revealed that the heat up time was 2 mins 1 s and the F0 value was 102.7. The present study suggests that the wireless thermologger system is useful in the management of the autoclave unequipped with sensor ports when it undergoes mechanical repairs. PMID- 14765557 TI - The effect of lipoprotein-associated cyclosporine on drug metabolism and toxicity in rats. AB - PURPOSE: The objective of the study was to examine the effect of lipoprotein associated cyclosporine on hepatic metabolism, hepatic lipoprotein receptors, and renal toxicity in comparison to the current commercially available cyclosporine (CSA) product. METHODS: Rats within the same group were given one of the following treatments: 10 mg/kg of CSA, plasma-CSA, very low-density lipoprotein (VLDL)-CSA, low-density lipoprotein (LDL)-CSA, LDL, high-density lipoprotein (HDL)-CSA, 1 mL/kg of vehicle, or saline intravenously for 14 days. Urine and blood samples were evaluated for renal function. Hepatic microsomes were prepared for immunoblotting and in vitro catalytic assays of CYP activity. Reverse transcription polymerase chain reaction (RT-PCR) was used to examine genetic regulation. RESULTS: (1) There were no statistical differences in cholesterol levels in lipoprotein-associated CSA groups as compared with vehicle controls. (2) A significant decrease in creatinine clearance was seen in the plasma-CSA treated group (56%; P < 0.05). (3) No suppressions of CYP3A protein, activity or mRNA were found in the VLDL-CSA treated group. (4) CYP3A mRNA was suppressed to a greater degree in the LDL- and HDL-CSA treated groups as compared with the suppression caused by CSA alone. (5) A significant suppression of hepatic low density lipoprotein receptor (LDL-R) mRNA levels was found in the LDL-CSA (50%; P = 0.0333) and plasma-CSA (40%; P = 0.1138), which was not attributed to LDL alone. (6) Significant suppression of scavenger-receptors class B type I (SR-BI) mRNA levels was found in the plasma-CSA group, although no significant differences in SRBI protein levels were seen between groups. CONCLUSIONS: Specific lipoprotein-CSA complexes appear to alter metabolic responses differently in comparison to CSA alone, indicating that the metabolism of CSA is dependent on the in vivo disposition of lipoprotein-CSA. Furthermore, LDL-R is one regulatory factor responsible for altering CSA metabolism as a result of an increase in uptake of CSA into hepatocytes. PMID- 14765558 TI - A method for demonstrating appropriate environmental protection for capping aseptically filled and plugged vials. AB - Conventional aseptic filling of drug product into vials ultimately includes a step that requires time and travel from the stoppering stage to the capping/crimping stage. Lyophilized product necessarily includes a partial stoppering step and, therefore, Grade A conditions. The level of environmental control that is required to protect the contents of fully stoppered, but uncapped, vials of sterile product from microbial contamination hasn't been formally defined. There is considerable range of opinion regarding this topic. This is the result of the lack of data that indicates the level of protection afforded by vials that are closed with a stopper, but that have not yet been capped. In order to provide some insight into the level of environmental control that is appropriate for vials at this stage of production, studies were conducted that consisted of exposing stoppered, uncapped vials containing sterile microbiological media to an aerosolized microbial challenge. The test samples were selected and assembled in such a way that a range of container/closure presentations was simulated. Furthermore, a microbial challenge was selected that was in excess of the environmental conditions to which product vials would be exposed. The results of this testing indicated a high level of confidence that stoppered, uncapped vials exposed to microbiologically challenging conditions would be expected to maintain the sterility of their contents. PMID- 14765559 TI - Characterization methods for the physical stability of biopharmaceuticals. AB - The characterization of protein aggregation and gelation using various methods, including scanning and transmission electron microscopy, small angle X-ray scattering, static light scattering, dynamic light scattering. infrared spectroscopy, Raman spectroscopy, rheology, and size exclusion chromatography (coupled with on-line laser light scattering, refractive index, and ultraviolet absorbance detection), are described in terms of theory and applications. PMID- 14765560 TI - Further development of the Temperament and Character Inventory. AB - The Temperament and Character Inventory is an internationally used personality questionnaire based on Cloninger's psychobiological theory of personality. Given some limitations of Version 9 a revised version was developed. The structural equivalence of the two versions was demonstrated from a cross-cultural perspective with 309 and 173 healthy volunteers from Sweden and Germany, respectively, who completed both versions in one session. In testing for the replicability of the factors across both samples as well as across both versions, an orthogonal Procrustes rotation method was used. The reliability coefficients for the revision were higher than the former version for both samples. The factor structures of the inventory remain highly equivalent across cultures and across versions. The results indicate a cross-cultural transferability of the Temperament and Character dimensions of the inventory. The stability and the validity of the 7-factor model of personality, as suggested by Cloninger, are supported. The Temperament and Character Inventory-Revised represents an important and useful method for the assessment of personality. PMID- 14765561 TI - Alexithymia and incidental learning of emotional words. AB - Alexithymia is thought to reflect a deficit in the cognitive capacity to process emotions. Prior research suggests that emotional valence has a memory enhancing effect in poor conceptual learning conditions. This study addressed the question of whether incidental learning of emotional words is a function of alexithymic tendencies. Incidental learning is unintentional learning that results from other activities. The 20-item Toronto Alexithymia Scale (TAS-20) and measures of depression and verbal intelligence were administered to 30 nonclinical subjects (15 women, 15 men) whose mean age was 35.5 yr. (SD = 8.6) along with a sequential word-word evaluation task. Partial correlations indicated that the TAS-20 subscale, Difficulties identifying feelings was negatively correlated with recall of positive distractor words but not with recall of neutral distractors or recall of positive or negative target words. Emotional valence appears to have less organizational power in the memory of individuals with difficulties in recognizing their feelings. PMID- 14765562 TI - Free will, determinism, and punishment. AB - Determinists were compared with weak, moderate, and strong libertarians with respect to philosophy of punishment. Data provided support for the contention that determinists are less punitive than libertarians. PMID- 14765563 TI - Risks of the consumption of beverages containing quinine. AB - Although the United States Food and Drug Administration banned its use for nocturnal leg cramps due to lack of safety and efficacy, quinine is widely available in beverages including tonic water and bitter lemon. Numerous anecdotal reports suggest that products containing quinine may produce neurological complications, including confusion, altered mental status, seizures, and coma, particularly in older women. Psychologists need to inquire about consumption of quinine-containing beverages as part of an evaluation process. PMID- 14765564 TI - Preliminary estimates of effects of a cognitive behavioral treatment system on drop-out rates in exercise programs at three YMCA facilities. AB - The effect of a treatment system based on principles of social cognitive and cognitive behavioral theory was estimated through case studies conducted in three YMCA fitness facilities. Significant (p < .05) reductions in drop-out rates of 23% (Georgia sample; n = 171), 27% (California sample; n = 159), and 21% (Florida sample; = 83) were found for the new and returning exercisers over the 5- and 6 mo. study time frames. Age was not related to drop-out rates in the treatment groups. Findings were consistent with previous research on earlier versions of the present treatment system, conducted with larger sample sizes within different fitness and wellness venues in the USA, U.K., and Italy. Replication, more detailed assessment of outputs of exercise, and tests of further revisions of the protocol were suggested to extend this preliminary research. PMID- 14765565 TI - Happiness of Asian Americans. AB - Past surveys found a positive relation between job satisfaction and socioeconomic status, with Asian Americans scoring low and African Americans and Euro-Americans scoring higher. As job satisfaction is a component of happiness, the question arises whether this relationship holds for happiness in general. Responses of a sample of 499 Asian Americans, 24,432 Euro-Americans, and 2,828 African Americans were analyzed. For both sexes, Asian Americans rated happiness significantly higher than African Americans. The rated happiness of Asian American and Euro American men was not significantly different, but Asian-American women rated happiness significantly lower than Euro-American women. Mean differences were less than one point. PMID- 14765566 TI - Visual perspective in causal attribution, empathy and attitude change. AB - The aim of the present study was to test the effect of visual perspective on the actor-observer bias. For this aim, we examined the effects of different visual perspectives on individuals' external and internal attributions. In addition to this, we examined the presence or absence of an attitude change toward the death penalty due to participants' visual perspective. One week before the experiment, we measured the participants' attitudes toward the death penalty. Then, during the experiment, films produced by one of the authors of this study were shown to two separate groups of participants. There were two films, each film constituting one of the two levels of visual perception. The content of each film was the memories of a person who was given the death penalty for the murder of his own brother. Level of visual perception was manipulated by using different camera perspectives, one from the actor's point of view and the other from the observer's point of view. At the end of the experiment, participants' attitudes toward the death penalty were measured again. PMID- 14765567 TI - Appropriate presentation of skin conductance accuracy with the Guilty Knowledge Test. AB - It was suggested that, when the Guilty Knowledge Test uses two repetitions, these should be combined showing the contribution of the second repetition to the accuracy of the first. Also recommended were effect size statistics in addition to accuracy rates. PMID- 14765568 TI - Parental perfectionism and children's test anxiety. AB - This study investigated the association between parental perfectionism and students' test anxiety, as measured on self-report questionnaires. A random sample of 90 students in their last two years of secondary schooling and their parents (81 mothers, 76 fathers), with approximately equal numbers of people from different socioeconomic backgrounds, were included. A significant contribution to anxiety associated with an examination was by maternal perfectionistic attitudes. Girls had higher scores on Test Anxiety than boys. The results are discussed in terms of the relationship of Test Anxiety to family variables, and directions for research are indicated. PMID- 14765569 TI - Predicting the outcome of methadone maintenance treatment with the Negative Treatment Indicators content scale from the MMPI-2. AB - Scores on the Negative Treatment Indicators content scale from the MMPI-2 were not significantly correlated with 5 measures of treatment outcome among 108 male patients on methadone maintenance. These results and those from other studies using this content scale as an independent variable with substance abusers suggest that the scale has yet to demonstrate consistent predictive validity with this population. PMID- 14765570 TI - Liebowitz Social Anxiety Scale: the Turkish version. AB - The present study aimed at establishing the Turkish version of the Liebowitz Social Anxiety Scale. For this aim interrater, test-retest, and internal reliability coefficients of the Turkish version were assessed as well as its convergent, discriminant, and criterion validity. Subjects were 88 patients with the diagnosis of generalized social phobia, 40 patients with a diagnosis of anxiety disorders other than generalized social phobia, and 40 people had no diagnosis of psychopathology. All reliability assessments indicated good reliability. For convergent and discriminant validity, correlations of 3 scores (2 subscales and the whole scale) and the Beck Anxiety Inventory were examined. Consistent with the expectations of the discriminant validity, correlations based on the whole sample were not significant. On the other hand, consistent with the expectations of the convergent validity, when patients with the diagnosis of generalized social phobia were considered, all the correlations were significant. Finally, the criterion validity of the Turkish version indicated that all 3 scores discriminated patients with the diagnosis of generalized social phobia, not only from the people who were free of psychopathology but also from the patients with the diagnosis of anxiety disorders other than generalized social phobia. Findings were discussed in the light of literature. PMID- 14765571 TI - Measuring market orientation: further evidence on Narver and Slater's three component scale. AB - A mail survey of a national random sample of 2,000 marketing managers was conducted. The data provided by 222 respondents were analyzed to assess the dimensionality of Narver and Slater's 15-item measure of market orientation. A confirmatory factor analysis, using LISREL 8.53, provided support for each of the separate dimensions of customer orientation, competitor orientation, and interfunctional coordination. However, a combined 3-factor model of market orientation was not supported. Directions for research are suggested. PMID- 14765572 TI - Using a pants alarm for the treatment of day wetting: problems with compliance. AB - A treatment of day wetting was evaluated in four normally capable children (ages 4:9, 4:10. 6:3, and 10:1 years). The treatment combined the use of a pants alarm with rewards (praise and tokens) for urinating in the toilet. The treatment was effective for one child only. The others showed many instances of noncompliance (not wearing the alarm, wearing the alarm with loose contacts or with the contacts on places where they could not detect urine). Tactics for dealing with this are suggested. PMID- 14765573 TI - Responsibility, guilt, and decision under risk. AB - We hypothesize that individuals' choices (risk-seeking/risk-aversion) depend on moral values and, in particular, on how subjects evaluate themselves as guilty or as victims of a wrong rather than on the descriptions of the outcomes as given in the options and evaluated accordingly as gains or losses (framing effect). People who evaluate themselves as victims are expected to show a risk-seeking preference (context of innocence). People who evaluate themselves as guilty are expected to show a risk-averse preference (context of guilt). Responses of 232 participants to a decision problem were compared in four different conditions involving two story formats (innocence/guilt) and two-question-options formats (gain/loss). Regardless of the format of the question options, the story format appears to be an important determinant of individuals' preferences. PMID- 14765574 TI - Note on Knight's analysis of the WAIS-III instruction effect on the Matrix Reasoning subtest. AB - Knight's 2003 analysis of the effect of the WAIS-III instructions on the Matrix Reasoning subtest was based on multiple t tests, which is a violation of conventional statistical procedures. Using this procedure significant differences were found between the group who know the subtest was untimed versus the group which did not know if the subtest was timed or untimed. Reanalysis of the data used three statistical alternatives: (a) Bonferroni correction for all possible t tests, (b) one-way analysis of variance, and (c) selected t tests with the Bonferroni correction. All three analyses yielded nonsignificant differences between means, thereby changing the conclusions of Knight's study. PMID- 14765576 TI - "Justification of effort" in rats: effects of physical and discriminative effort on reward value. AB - The justification of effort is the tendency for humans to justify the amount of work they have put into an achievement by increasing the perceived value of that achievement after the effort has been expended. A positive effect of effort on reward value has also been reported for pigeons and starlings. The present study examined the effects of different amounts of required physical and discriminative effort on primary reward value in rats. On each day of training, rats underwent one high-effort training session and one low-effort training session in an operant chamber to earn either a grape-flavored reward pellet or a bacon-flavored reward pellet. Half of the rats exerted high effort to earn the grape pellets and low effort to earn the bacon pellets, with the arrangement reversed for the other half. On each test trial, each rat had the opportunity to consume 3 pellets of each flavor at the choice point in a T-maze. Analysis indicated no significant difference between the frequency of high-effort flavor choices and low-effort flavor choices. A positive effect of effort on reward value was not demonstrated in this experiment. PMID- 14765575 TI - Pastoral response to the Clinton-Lewinsky affair in conservative and liberal churches. AB - The study examined what factors led pastors to say something either negative or positive about former President Clinton's affair with Monica Lewinsky for a sample from "Middletown." There was no significant difference between three Protestant categories (Holiness/Pentecostal, white conservative Protestant, and mainline Protestant) in whether ministers said something or not. Analysis for those who said something indicated that the ministers who made the most critical statements in their sermons were from the most conservative/orthodox of our category of churches, i.e., Holiness/Pentecostal congregations, from the strictest churches and from churches with the greatest number of Republicans (betas were .30, .26, and .19, respectively). Also, members of Holiness/Pentecostal churches were more likely to be working class but not more Republican. Possible explanations are discussed. PMID- 14765578 TI - Depression in acute coronary syndrome. AB - This study examined the prevalence of depression based on scores of 200 patients with acute coronary syndrome on the Emotion Profile Index of Plutchik and its relationship with the type of acute coronary syndrome and the severity of ischemic heart disease. Patients with acute coronary syndrome scored higher on depression than the control group. There was no difference in scores on Depression by type of acute coronary syndrome and no significant mean differences on Depression for patients with and without left ventricular failure. Patients with acute myocardial infarction and ventricular fibrillation scored lower on Depression than other patients with acute myocardial infarction and control group. This study supports the view that patients with acute myocardial infarction and ventricular fibrillation and lower scores on Depression have good prognosis during hospitalization and maybe for the long term. PMID- 14765577 TI - Adequacy of an English version of the Kuwait University Anxiety Scale based on back translation and testing bilingual subjects. AB - An English version, a translation of the Kuwait University Anxiety Scale written originally in Arabic, was prepared. Several cycles of translation and back translation were carried out. Bilingual Arab college students, studying English language and literature (N=74) responded to the two versions of this scale in counterbalanced order. The correlation between the two forms was .96 denoting high cross-language equivalence. The nonsignificant mean difference for total scores between the two versions was an indication that they functioned as equivalent stimuli. Thus, the English version of the scale is highly recommended for use with English-speaking subjects. PMID- 14765579 TI - The Kuwait University Anxiety Scale: a cross-cultural evaluation in Kuwait and United States. AB - Kuwaiti (n=460) and American (n=273) college students responded to the Kuwait University Anxiety Scale in Arabic and English languages, respectively. (Coefficients alpha were .93 and .95, while the item-remainder correlations ranged from .37 to .75 and from .33 to .81 for the Kuwaiti and American samples, respectively. Three factors were identified for the Kuwaiti sample: (1) Cognitive/Affective, (2) Somatic, and (3) Behavioral/Subjective Anxiety and two factors for the American sample: (1) Cognitive/Affective/Behavioral and (2) Somatic Anxiety. The Kuwaiti sample had significantly higher mean scores on 13 of the 20 items and on the total scale score than the American sample. Sex differences were significant in both countries. By and large, the Kuwait University Anxiety Scale seems viable in the American and Kuwaiti contexts. PMID- 14765580 TI - Multimethod validation of the Beck Depression Inventory-II and Grossman-Cole Depression Inventory with an inpatient sample. AB - Psychometric examination of the Beck Depression Inventory-II has been adequate and varied for outpatient and nonclinical samples, but is still comparatively scant for inpatient psychiatric samples. In particular, important factor analytic questions have not been assessed on inpatient samples at all. Given that over time alone (regardless of symptom change) test scores on depression measures can spuriously drop, a convergent psychometric analysis of the Beck Depression Inventory-II was undertaken with a new measure of depression severity, the Grossman-Cole Depression Inventory. A sample of 101 psychiatric inpatients was administered both tests. Item and scale level psychometrics were reviewed for each measure. Results were highly consistent with previous findings, providing support for the use of the Beck Depression Inventory-II with inpatient samples. Moreover, the new inventory performed quite well, suggesting it has promise as another measure for self-reported depression symptom severity. The Beck Depression Inventory-II appears appropriately considered as a single-factor hierarchical measure of depression with the current inpatient sample. Moreover, the Grossman-Cole Depression Inventory is related enough to the Beck inventory to provide adequate similarity without being so similar the measures duplicate each other. PMID- 14765581 TI - Hobfoll's strategic approach to coping: reliability and validity of the Italian adaptation. AB - A brief review of Hobfoll's Conservation of Resources theory is given, and the Italian adaptation of a measurement, based on this theory, the Strategic Approach to Coping Scale, is presented. The scale is useful measure to assess stress and coping conceptualized to identify coping styles. The investigation used a sample of Italian people (N=251, 18 to 25 years of age). Confirmatory factor analysis provided support for the validity of the scale. Data on the Italian adaptation are discussed. PMID- 14765582 TI - Examination stress, salivary cortisol, and academic performance. AB - Immediately before and after participation in a written final examination, 11 graduate students rated their self-perceived stress and provided saliva samples for cortisol assay. Students rated stress higher before the examination, and these ratings were associated with increased salivary cortisol. Students who reported higher stress and had higher cortisol levels before the examination tended to have significantly lower examination scores. PMID- 14765583 TI - Effects of situational conditions on students' views of business ethics. AB - This study investigated undergraduates' responses regarding selected ethical issues facing managers and employees of today's businesses. The focus of the study lies in the influences of two situational variables (organizational roles and prospects) on students' response pattern. Japanese students (306 men and 81 women, M = 20.1 yr., SD = 2.2) imagined that they were managers or operative employees of a middle-sized manufacturing company and that their company had high or low prospects. The response pattern tended to be more ethical for "managers," whereas the response pattern tended to be less ethical for "employees" in a "low prospect" than in a "high prospect" company. PMID- 14765584 TI - Relationship between God-image and religious behaviors. AB - This study examined the relationship between God-image and religious behaviors. Participants were 344 undergraduate students at a private midwestern university. Analysis showed strong correlations among three aspects of God-image (presence, acceptance, and challenge) and six items that measured religious practices and religiously motivated volitional behaviors. The findings held when data were controlled for racial and religious diversity. As such, these results differed from an earlier study indicating religious practices to be independent of God image. PMID- 14765585 TI - Cognitive styles and learning preferences of undergraduate business students in North Cyprus. AB - Measures of cognitive style and learning preferences were administered to 127 (75 men and 52 women) Turkish undergraduate business students. Students with analytical cognitive styles were more likely to prefer teacher-dependent and collaborative learning settings. As expected the mean Cognitive Style Index scores for the current Turkish sample was higher (more analytical) than those for similar western samples reported in the literature. Furthermore, students who had completed more credits towards the completion of their degrees had a higher mean score. The hypothesis regarding differences between scores of men and women was not supported. PMID- 14765586 TI - Evaluating an all-ranks military staff ride. AB - Outcomes for a staff ride to a Missouri Civil War battlefield by enlisted personnel and officers of the Army Reserve were evaluated. As expected, enlisted personnel and junior officers were much less likely to have participated in a staff ride previously. Nearly all soldiers expressed an interest in going on another all-ranks staff ride, and a statistically significant gain was achieved in expressed interest in American Civil War history. Most personnel agreed or strongly agreed with the concept of an all-ranks staff ride. There were few differences among officers and enlisted personnel in the success of the outcomes evaluated. However, learning seemed to be maximal among those with an intermediate level of previous staff ride experience. PMID- 14765587 TI - Do American citizens consider their food supply at risk for terrorism attack? AB - Since the attacks on 9/11, 2001 Americans have been told to be alert and observant. The purpose of this research was to investigate if Americans feel that the food supply is in danger and if their food habits have changed in any way due to the traumatic events. The results indicate that very few individuals in this survey have actually changed their food habits since 9/11. However, a large percentage of our surveyed group feels that the food supply is at risk. PMID- 14765588 TI - Schemata and attitudes toward persons with intellectual disability in Japan. AB - 375 parents of pupils were asked to respond to 14 questions on attitudes toward persons with intellectual disability. About 30% of variance in attitude was explained by a set of variables. The effects of schemata concerning person with intellectual disability had a great effect on attitudes toward such persons. The guess of a greater contribution of heredity as the basis of intellectual disability and of fewer such persons in the future of their own families than in the general population was associated with greater negative attitudes toward persons with intellectual disability. As these estimations relate to teachable information, the attitudes might be improved. PMID- 14765589 TI - Homosexual parents in custody disputes: a thousand child-years exposure. AB - Through 1998, 78 custody appeal decisions involving 79 homosexual parents were evaluated. The 142 children involved in these cases were exposed to a thousand child-years of homosexual parenting. In appeals records (a) parents recorded as lying or engaging in criminality or homosexuality were more apt to be recorded as harming children; (b) homosexuals more frequently were recorded as lying or engaging in criminality; (c) in 54 (70%) disputes the homosexual parent or his associates and in 4 (5%) the heterosexual parent was recorded as having exposed the children to harm(s), e.g., neglect, seduction; and (d) homosexuals were responsible for 111 (97%) of the 115 recorded harms to children. For 78 nonhomosexual vs nonhomosexual control cases, in 11 (14%) the 141 children were exposed to 12 harms. PMID- 14765590 TI - Involuntary sex experienced by homeless young people: a public health problem. AB - The incidence of involuntary sex among homeless young people is considerably higher than in the general population. The most common reason for unwanted sex is being drunk or high at the time. There is a need for programs that provide homeless young people with knowledge and decision-making skills to enable them to avoid unwanted sex and thus exposure to adverse sexual health outcomes. PMID- 14765591 TI - Contemporary university students' ratings of characteristics of men, women, and CEOs. AB - Very few women have reached the highest echelons of corporate America, perhaps because gender stereotypes, including perceptions of women that vary from those of successful executives, block their promotion and advancement. In the current study, differences in how participants perceive similarities in characteristics of successful executives and those of both men and women were studied. The scope of the extant program of research is also extended upward in the organizational hierarchy with the operationalization of executive as "CEO" (Chief Executive Officer) rather than as "manager" or "middle-manager." While men in general continue to be likened more to successful executives than do women in general, the gaps between male and female CEOs' similarities and between successful male and female CEOs' similarities to prototypically successful executives were smaller than reported in the 1970s. Noteworthy trends regarding 92 characteristics from Schein's Descriptive Index are also discussed. PMID- 14765592 TI - A recursion formula for Bayesian probabilities. AB - A recursion formula for Bayes' formula is derived. The formula is useful in applications in which diagnoses are added in a stepwise way to predict a criterion. On each step, changes in various diagnostic measures can be easily evaluated. PMID- 14765593 TI - Construct validity of the Adolescent Resilience Scale. AB - The aim of this study was to assess the construct validity of the Adolescent Resilience Scale which measures the psychological features of resilient individuals. Research involving this scale, the Negative Life Events Scale, and the General Health Questionnaire was conducted with a group of 207 Japanese undergraduate students (104 men and 103 women; M age=20.2 yr., SD=.9). A cluster analysis for the Negative Life Events Scale and General Health Questionnaire yielded three clusters: (1) mentally healthy with little experience of Negative Life Events, (2) poorer mental health with many experiences of Negative Life Events, (3) mentally healthy despite many experiences of Negative Life Events. These three groups were defined as (1) Well Adjusted, (2) Vulnerable, and (3) Resilient, respectively. Mean differences in scores on the Adolescent Resilience Scale among the three groups were subjected to one-way analysis of variance. The mean scores of both the Well Adjusted and Resilient groups were higher than that of the Vulnerable group, and therefore support the construct validity of the Adolescent Resilience Scale. PMID- 14765594 TI - Age-related parenting stress differences in mothers of children with spina bifida. AB - This study examined whether ages of child and parent were risk factors for general parenting stress and disability-specific stress in families of children with spina bifida. Parents of 64 children with spina bifida completed the Parenting Stress Index-Short Form, Parents of Children with Disabilities Inventory, and measures of family support and resources. Scores of families with children under 6 years (preschool) versus 6- to 12-yr.-old children (school age) were compared, as were scores of mothers above or below Age 35. Parents of school aged children reported significantly higher stress on the Concerns for the Child domain of the Parents of Children with Disabilities Inventory. Mothers over 35 tended to report higher stress in the Concerns for the Child and Medical/Legal Concerns domains of the Parents of Children with Disabilities Inventory. No associations with medical severity, socioeconomic status, family resources, or family support were detected. As the children age and disability-related differences become more apparent, the same level of functioning and severity of disability may be associated with additional parenting stress. Older mothers and those with school-age children may need more resources than current social support systems typically provide. PMID- 14765595 TI - Severity and internal consistency of self-reported anxiety in psychotic outpatients. AB - To assess the severity of self-reported anxiety in psychiatric adult outpatients (> or = 18 yr. old) who were diagnosed with psychotic disorders, the Beck Anxiety Inventory was administered to 55 (50%) women and 55 (50%) men who were diagnosed with paranoid schizophrenia, schizoaffective, or delusional disorders. The internal consistency of the scores was high (coefficient alpha=.92), and the scores were not significantly correlated with sex, being Euro-American, or age. Furthermore, the mean cores of the three diagnostic groups were comparable. Based on the interpretive cut-off score guidelines given in the manual, 24% of the patients were mildly anxious, 22% were moderately anxious, and 18% were severely anxious. The results are discussed as indicating that there is a high prevalence of self-reported anxiety in outpatients who are diagnosed with psychotic disorders. PMID- 14765596 TI - Brief report on predictive validity evidence of global risk indicators in the lives of court-involved youth. AB - Previous work has reported the psychometric properties and evidence for concurrent validity of a battery of items measuring global risk indicators in the lives of court-involved adolescents. The present study reports additional data on the predictive validity of this battery in a sample of 224 families of adolescents who were assessed by intake workers in a juvenile court and subsequently referred for services. Analysis yielded significant differences between groups of youth who were and were not referred to more intensive mental health-based treatment on all domains of risk in his battery. PMID- 14765597 TI - Correlations among symptoms of depression and problematic eating patterns in intercollegiate athletes. AB - Using self-report questionnaires, depressive symptoms as measured by the Beck Depression Inventory-II and problematic eating patterns as measured by the Eating Disorders Inventory were not significantly correlated for 57 (39 women) Division I varsity intercollegiate athletes. PMID- 14765598 TI - Reading comprehension: think and know verbs. AB - Verbs such as think, know, remember, and guess play a pivotal role in understanding, monitoring, and transformation of internal states. We focus on the specific words as think and know, polysemous cognitive verbs that show hierarchical organization and high frequency of use in children's and adults' lexicons. According to Booth and Hall's model, think and know present a conceptual organization that involves low conceptual levels (perception, memory, comprehension) and high conceptual levels (evaluation, metacognition, planning). The aim of this research was to study the relationship between children's comprehension of text processing and the conceptual levels of the above-described verbs. The research concentrated on 9-, 11-, and 13-yr.-old children's ability to understand think and know. Analysis yielded a strong relation between knowledge at the conceptual level of these verbs and reading comprehension skills. Moreover, it highlights the importance of this linguistic competence in skilled readers. PMID- 14765599 TI - Correlational analysis of MicroCog: Assessment of Cognitive Functioning with the Wechsler Adult Intelligence Scale-III for a clinical sample of veterans. AB - With rapid advances in technology and an emphasis in efficiency in psychological testing, there is a need to investigate the relation between a computerized cognitive examination and a traditional individually administered intelligence test. The current study provided correlations of intelligence scores from MicroCog: Assessment of Cognitive Functioning and intelligence scores from WAIS III. MicroCog is a single computerized test measuring intelligence and the WAIS III is a single traditionally administered test measuring intelligence. The study included 30 participants referred for psychological testing at a Veterans Medical Center in Tennessee as part of the standard intake process. Half of the participants were administered MicroCog first and half the WAIS-III first. Analysis indicated scores on the two tests were positively correlated. Index scores measuring similar constructs also were positively correlated. However, MicroCog was significantly more difficult than the WAIS-III in that scores were, on average, lower on MicroCog. PMID- 14765600 TI - Dimensionality of the Kansas Family Strengths Scale and the Kansas Marital Satisfaction Scale as revised to capture changes in marital satisfaction. AB - The Kansas Marital Satisfaction Scale was modified to reflect issues of relationship development and administered, along with 19 items from a previously reported Kansas Family Strengths Scale by Schumm, Bollman, Jurich, and Hatch, to 337 married couples from a midwestern urban community. High internal consistency was obtained for the revised version (alpha=.93), and the dimensionality of the Kansas Family Strengths Scale was approximately similar to that reported previously. In both this and the previous study, communication and conflict resolution items factored together while items for enjoying time together and actually spending time together factored separately. Likewise, in both studies, normative and affective commitment factored separately, and the internal consistency of the affective commitment subscale exceeded that for the normative commitment subscale. While the subscales for affective commitment, communication or conflict resolution, positive interaction, and time together yielded fair to adequate internal consistency in both studies, both studies indicated very low internal consistency for the normative commitment subscale, suggesting a need for improvement in that particular subscale. PMID- 14765601 TI - An application of individual subtest scores calculation in the Cantonese version of the Test of Everyday Attention. AB - This study aimed to apply Crawford, et al's formula for calculating individual subtest scores of the Cantonese version of the Test of Everyday Attention. A total of 133 (72 men, 61 women) healthy Hong Kong Chinese were recruited from the general public. The sample reported a mean age and education of 35.2 yr. (SD=10.2) and 11 yr. (SD=3.1), respectively. Tables for examining whether an individual's subtest profile contains reliable and abnormal subtest discrepancies are presented and discussed. The data are useful for clinicians when they take into account cultural differences of Cantonese-speaking clinical populations. PMID- 14765602 TI - Weighted specific-category kappa measure of interobserver agreement. AB - When two observers classify a sample of items using the same categorical scale, and when different disagreements are differentially weighted, the weighted Kappa (Kw) by Cohen may serve as a measure of interobserver agreement. We propose a Kappa-based weighted measure (K(ws)) of agreement on some specific category s, with Kw being a weighted average of all K(ws)s. Therefore, while Cohen's Kw is a summary measure of the overall agreement, the proposed K(ws) provides a measure of the extent to which the observers agree on the specific categories, with both measures being suitable for ordinal categories because of the weights being used. Statistical inferences for K(ws) and its unweighted counterpart are also discussed. A numerical example is provided. PMID- 14765603 TI - Themes observed in early Qur'anic material: a brief commentary on Michael Sells' text, Approaching the Qur'an. AB - In 1999 Sells described and explained the early revelations to Muhammad in the Qur'an. Those revelations are analyzed for thematic content, primarily to stimulate student discussion in classes using Sells' text. Four primary themes are discussed; the theme most frequently cited is the Day of Reckoning or Judgment. Living a righteous life, being aware that unbelievers may prosper for a time, and remembering God are also mentioned very often. Issues for students using Sells' text in university classes are discussed. The possibility of considering Jesus as a type of Karim (Islamic generous folk hero) is considered. PMID- 14765604 TI - Action models and verbal descriptions in object representations given through gestures by preschool children. AB - Previous research suggests that young children have difficulty producing pantomimic gestures with imagined objects for which no substitute objects are presented. They frequently substitute specific body parts to represent the object through gestures involved in the action. This study examined whether adults' verbal descriptions or symbolic action models help to produce an imaginary-object gestural response. 53 children (26 4-yr.-olds, and 27 5-yr.-olds) performed gestural tasks in which they were asked to pretend to use common objects, e.g., "pretend to brush your teeth with a toothbrush." They were then given verbal descriptions or action models to perform those same tasks. Analysis indicated that providing verbal descriptions and action models helps to produce an imaginary-object response in 4- and 5-yr.-old children. Action models proved more effective than verbal descriptions among 4-yr.-old children. Results are discussed indicating that each verbal description and action model could be influenced by different mechanisms underlying preschool children's symbol production. PMID- 14765605 TI - Anticipating the correct matching response in a second-order matching-to-sample task. AB - A study was designed to evaluate the effects of anticipating the matching response on acquisition and transfer of a second-order matching-to-sample task. 40 college students of both sexes were assigned to different anticipation procedures that involved several verbal descriptions of the correct response, as well as others not involving explicit descriptions. All subjects were exposed to a pretest, two training blocks, two transfer test blocks, and a posttest. Anticipation procedures were evaluated during the first training block using an observational training procedure, except for 8 subjects, who were exposed to two training control procedures. Analysis showed acquisition and transfer of successful matching seems to depend on a verbal discrimination of the matching criterion exemplified by second-order stimuli. This discrimination was facilitated by prior conditions, as when subjects showed correct matching performance at pretest or was established with anticipation procedures that required explicit reading of verbal descriptions of the response choice. PMID- 14765606 TI - Regulation of emotional security by children after entry to special and regular kindergarten classes. AB - In this paper early adaptation after children's entry to kindergarten is conceptualized as a process of achieving emotional security. It was hypothesized that children adapt to school by means of security seeking from the teacher and behavioral inhibition. 30 normal children from regular classes and 36 children with a variety of problems, e.g., behavioral, emotional, and family problems, from special classes were rated by their teachers on the Inhibition Scale and Security Seeking Scale on 5 occasions during the first 3 mo. at school. By the end of this period teachers judged the intensity of behavior problems on the Internalizing Problem Scale and the Externalizing Problem Scale. Analysis showed that initial high scores on the Security Seeking Scale and Inhibition Scale decreased sharply during the first weeks, and that children from special classes scored consistently higher on the Security Seeking Scale and more variable on the Inhibition Scale than children from regular classes. Girls had higher scores than boys on both scales. Recent stress in the family as rated by the teacher was positively related to both scores on the Inhibition and Security Seeking Scales after entry. Finally, scores on the Security Seeking and Inhibition Scales over the first three months predicted scores on the Internalizing Problem Scale by the end of this period, especially for children in special classes. We conclude that understanding adaptation after school entry as a process of obtaining emotional security is productive, providing a means to link entry behavior to precursors and consequences. PMID- 14765607 TI - A re-analysis of Price's "Islam and human rights: a case of deceptive first appearances". AB - Daniel Price in his analysis of Islamic Political Culture and Human Rights concluded that "... government rooted in Islam does not facilitate the abuse of human rights." A re-analysis of his data for 23 Islamic governments demonstrates otherwise. There is a significant trend (p<.03), despite the low statistical power available in only 23 cases, for an inverted quadratic relationship between Islamic Political Culture and Human Rights. Among the nations scoring low on Islamic Political Culture, the correlation between the two variables is -.01 (ns); among those scoring high on Islamic Political Culture, the correlation shifts to -.78 (p<.02). At lower scores for Islamic Political Culture, there may indeed be little relationship between Political Culture and Human Rights; however, at higher scores there appears to be a significant relationship between increasing Islamic Political Culture and a decline in Human Rights. The data suggest that extreme applications of Sharia law (if not any secular or religious legal system) may have serious implications for human rights--or at least, Western Euro-American conceptualizations of human rights. At the same time, support for human rights may increase as Islamic governments shift from mostly secular to moderate applications of Islamic law. PMID- 14765608 TI - Sex differences in relations of cardiorespiratory and mood changes associated with self-selected amounts of cardiovascular exercise. AB - 44 formerly sedentary men (n=22) and women (n=22) self-selected cardiovascular exercise intensities, durations, and frequencies for 12 wk. A mean of approximately 2 exercise sessions/week, for 28 to 29 min./session were completed, and small (ds=.30 to .35) but significant (p<.05) cardiorespiratory (VO2 max) improvements were found. Reductions in Depression and Tension scores on the Profile of Mood States were also significant (ps<.05) over the 12 wk. No sex differences were found for change on any of the variables of interest. Analysis showed no significant correlations between change on either Depression and Tension scores, and change in VO2 max, over the 12 wk. Limitations and the need for replication with a control group were discussed. The possible advantages of allowing new and returning exercisers to self-select initial physical activity amounts were discussed. PMID- 14765609 TI - Fluoride uptake by enamel and dentin from bonding agents and composite resins: a comparative study. AB - The present study was performed to compare the amount of in vitro fluoride uptake by enamel and dentin from fluoridated bonding agent with non-fluoridated composite (Group I), non-fluoridated bonding agent with fluoridated composite (Group II) and fluoridated bonding agent with fluoridated composite resin (Group III). Sixty extracted premolars were selected and divided into three groups of 20 teeth each. Restorative materials were applied according to manufacturer's instructions into standard windows created in the teeth, cured and placed in de ionised water for three months. The fluoride content of successive acid etch biopsy was determined by specific ion-electrode analysis. Although significant amount of fluoride uptake occurred in all the test groups, fluoride uptake was found to be highly significant in dentin when compared to enamel in Group, I, II and III (P<0.001). The fluoride uptake was greater by both enamel and dentin in Group III ie fluoridated bonding agent with fluoridated composite resin. PMID- 14765610 TI - Hypertaurodontism in molars and premolars: management of two rare cases. AB - The management of two cases of hypertaurodontism, is presented. The first case shows this uncommon dental anomaly in all the four permanent first molars. The second case is concerned with endodontic management of a hypertaurodontic mandibular second premolar. The authors wish to highlight the importance of careful radiographic examination in diagnosing this rare dental anomaly in view of the difficulties encountered in performing endodontic treatment. PMID- 14765611 TI - Relation of salivary inorganic phosphorus and alkaline phosphatase to the dental caries status in children. AB - The phosphorus concentration and alkaline phosphatase activity in saliva may be related to the susceptibility to dental caries. Thirty children in the age group of 4-6 years were divided into 3 groups. Group 1, children had rampant caries with dfs>10, group 2- dfs>5<10, group 3 caries free with dfs<1. The unstimulated mixed salivary sample was collected from each child and assayed by biochemical methods to determine the phosphorus and alkaline phosphatase concentration in saliva sample. The results showed that higher alkaline phosphatase activity and phosphorus concentration were associated with rampant caries. PMID- 14765612 TI - Radicular cyst associated with endodontically treated deciduous tooth: a case report. AB - This article presents a case report of radicular cyst associated with an endodontically treated deciduous second molar causing displacement of the permanent successor, with accompanying buccal expansion. Cystic sac was removed surgically along with the involved tooth under general anesthesia. Healing was uneventful. Histopathologically the cystic sac was consistent with the features of radicular cyst. Unusual amorphous, eosinophilic, atubular material incorporated within the cystic epithelium was observed. PMID- 14765613 TI - New insights into improving the oral health of visually impaired children. AB - Comprehensive dental care for 150 blind school children was carried out. The main goal of this oral hygiene programme was to create awareness to enable them to be independent in their oral hygiene care. PMID- 14765614 TI - Intraoral actinomycotic lesion: a case report. AB - Actinomycosis often referred to, as the chameleon of the head and neck pathology is a rare disease, uncommon in children. This article refers to a case of intraoral actinomycotic lesion of the palate in a child following a rare aetiology, the relevant literature, clinical course and its successful resolution. PMID- 14765615 TI - Prevalence of dental health problems among school going children in rural Kerala. AB - The purpose of this study was to know the prevalence and pattern of dental health problems in rural school children of Kerala and to identify the priority area for dental health education programmes. The children were examined and findings recorded. the findings show that more than 50% of the children in the 12 to 15 years of age group in rural Kerala suffers from some form of dental diseases. Males and females are equally affected and dental caries is the most common problem encountered. PMID- 14765617 TI - Cystic fibrosis in India. PMID- 14765616 TI - Herniation of the buccal fat pad into the oral cavity: a case report. AB - An interesting lesion is presented, wherein a tiny traumatic perforation of the buccal mucosa and buccinator muscle forced a large portion of the buccal fat pad to extrude into the oral cavity. Such a situation can alarm any clinician & reinforces the importance of careful history taking and thorough examination, before treating patients. PMID- 14765618 TI - Bone mineral health of Indians. PMID- 14765619 TI - Bone mineral parameters in healthy young Indian adults with optimal vitamin D availability. AB - BACKGROUND: Several recent studies indicate a marked prevalence of vitamin D deficiency in asymptomatic, apparently healthy urban subjects from different socioeconomic groups in north India. METHODS: To further examine this trend, we studied 40 men and 50 women, 20-30 years of age, from the Indian paramilitary forces. These individuals consume a nutritious, high-protein diet, have optimal exposure to sunlight and undertake strenuous outdoor physical exercise. RESULTS: The mean serum calcium, phosphorus and alkaline phosphatase levels were normal in both men and women. The mean (SD) serum intact parathyroid hormone and 25 hydroxyvitamin D3 levels were 19.3 (8.2) pg/ml and 18.4 (5.3) ng/ml in men, and 11.9 (6.6) pg/ml and 25.3 (7.4) ng/ml in women. Bone mineral density estimated in 20 men and 22 women revealed that in comparison with white Caucasians, 35%-50% of men and 14%-32% of women were osteopenic at different sites, while an additional 10% of men had osteoporosis of the lumbar spine. CONCLUSION: We found that with optimal nutrition, good sunlight exposure and regular physical exercise, healthy young individuals have normal bone and mineral biochemical values. The reasons for the abnormalities detected in bone mineral density in them needs further study. The impact of childhood nutrition on accumulation of peak bone mass may contribute to our findings. There is a need for establishing normative bone mineral density data for Indians. PMID- 14765620 TI - Symptoms related to the reproductive tract and mental health among women in rural southern India. AB - BACKGROUND: Symptoms related to the reproductive tract and symptoms of psychological distress are commonly seen in women living in South Asia. This study aimed to determine the association, if any, between symptoms related to the reproductive tract and mental health among women in rural southern India. METHODS: The study was conducted in a community setting of a comprehensive community health programme in a development block in rural southern India. It was a cross-sectional survey with a nested case-control study. The main outcome measure was the probability of mental distress as assessed by the Tamil version of the General Health Questionnaire-12 (GHQ). RESULTS: Of the 622 subjects recruited, 150 (24.4%) complained of at least one symptom related to the reproductive tract. The commonest symptom was vaginal discharge (17.5%). Significantly higher scores on the GHQ were associated with the presence of many current and past symptoms related to the reproductive tract, and receiving treatment for the same, even after adjusting for age, education, current marital status and type of house, using multiple regression analysis. GHQ scores were positively correlated with a higher number of current and past symptoms. Significantly higher scores were also associated with greater knowledge about prevention of sexually transmitted diseases and reproductive tract infections. Being currently married and educated were significant protective factors. A nested case-control study was done with the presence of a current common mental disorder as the outcome. Symptoms and other factors dated prior to this outcome were considered potential risk factors. Logistic regression models were developed. The presence of a symptom related to the reproductive tract for more than one month (OR 3.6; 95% CI 1.8, 7.2) and a history of similar symptoms over the past year (OR 2.2; 95% CI 1.3, 3.7) were risk factors for the development of common mental disorders. Higher education (OR 0.5; 95% CI 0.3, 0.7) and being currently married (OR 0.2; 95% CI 0.1, 0.6) were protective factors. CONCLUSION: The cross-sectional data suggest an association between symptoms pertaining to the reproductive tract and psychological distress. The results of the nested case control study suggest that symptoms related to the reproductive tract are risk factors for common mental disorders, while education and being currently married are protective factors. PMID- 14765621 TI - Distribution of C282Y and H63D mutations in the HFE gene in healthy Asian Indians and patients with thalassaemia major. AB - BACKGROUND: Mutations in the HFE gene have been shown to be strongly associated with hereditary haemochromatosis, an autosomal recessive disease of iron overloading. The majority of patients with hereditary haemochromatosis possess a homozygous mutation C282Y that disrupts the binding of the HFE gene with beta2 microglobulin and prevents its surface expression. Another HFE mutation H63D is known to increase the relative risk of developing hereditary haemochromatosis. This disease is rare in India although secondary haemochromatosis is commonly seen among children suffering from thalassaemia major. The status of HFE mutations has not been explored among Indians, particularly in patients with thalassaemia major. It is also possible that in India clinical haemochromatosis could be masked by iron deficiency. METHODS: We examined a cohort of 59 unrelated, healthy individuals from north India, 57 from south India and 75 thalassaemia major patients from north India for HFE mutations (C282Y and H63D) in cis/trans by the polymerase chain reaction sequence-specific primer method. RESULTS: The C282Y and H63D mutations in the HFE gene were rare among Indians. Although the HFE mutations were increased among patients of thalassaemia their effect on iron burden or disease pathogenesis remains unclear. CONCLUSIONS: Hereditary haemochromatosis is rarely observed among Indians and so are the C282Y and H63D mutations in the HFE gene. Long-term follow up studies would be required to determine whether the relatively higher frequency of these mutations among patients of thalassaemia has any influence on iron accumulation. PMID- 14765622 TI - The antiphospholipid syndrome. AB - The antiphospholipid syndrome encompasses a wide spectrum of presentations cutting across all subspecialties of medicine. It is characterized by recurrent thrombotic events involving both the arterial and venous systems. Large arteries and veins as well as the microcirculation are involved. Recurrent strokes, myocardial infarction, pulmonary embolism, gangrene of the digits, etc. cause much morbidity and mortality in affected patients. It is recognized as an important cause of recurrent pregnancy loss. The risk in pregnancy extends to a propensity towards pre-eclampsia, abruptio placentae and intrauterine growth retardation. It often manifests as asymptomatic thrombocytopenia and sometimes as a life-threatening form called catastrophic anti-phospholipid syndrome. The management of thrombotic events rests on high grade anticoagulation (INR 3-4) as lower values of INR than this often fail to prevent recurrence. Aspirin is generally added in case of arterial thrombosis. A combination of heparin and aspirin at least in the first trimester and sometimes throughout pregnancy is used to prevent foetal loss. PMID- 14765623 TI - Detecting lymph node metastases in prostatic cancer. PMID- 14765624 TI - Worms and satellites. PMID- 14765625 TI - Beyond DOtS: avenues ahead in the management of tuberculosis. AB - India has almost 30% of the global burden of tuberculosis (TB)--one person dies of the disease every minute in our country. India has mounted the second-largest DOTS programme in the world to control this disease. However, DOTS has its limitations and newer approaches have been developed over the years to overcome the global burden of tuberculosis. Problems with health facilities, patients, drugs and the disease itself constitute some of the hurdles in the implementation of the DOTS programme. In an attempt to go beyond DOTS, the WHO launched the 'Stop TB Initiative' in 1988. Against the background of irrational antituberculosis drug use, which contributes to increasing drug resistance, the effective involvement of private healthcare providers is imperative to achieve better geographical and patient coverage for the implementation of DOTS. The WHO is currently addressing the issue of involving private practitioners in tuberculosis control in a programme called Public-Private Mix DOTS (PPM DOTS). The Stop TB Initiative is also active in the area of dual infection with HIV and tuberculosis, and the initiatives that have been taken in this area include 'ProTEST', community contribution to tuberculosis care, and development and dissemination of training materials and guidelines. The DOTS-Plus strategy for the management of multidrug resistant (MDR)-TB and the establishment of the Green Light Committee to review project applications in this area are initiatives taken to curb the problem of drug resistance in tuberculosis. Even decades after the introduction of the DOTS strategy, much needs to be done to expand the services to the entire population; it is now essential to develop strategies that go beyond DOTS. PMID- 14765626 TI - Doctored admissions: are we sowing the right seed? PMID- 14765627 TI - Bottoms up. PMID- 14765628 TI - Ultrasound scanning in pregnancy: the hope and the shame. PMID- 14765629 TI - A matter of little importance. PMID- 14765630 TI - Interventional radiology: a new name? PMID- 14765631 TI - Noise pollution and its effect on workers of the steel and iron industry. PMID- 14765632 TI - The ethics of live operative workshops. PMID- 14765634 TI - Fusing traditional techniques and new technology. PMID- 14765633 TI - Polio eradication in India: some queries. PMID- 14765635 TI - Amelogenesis imperfecta: the multidisciplinary approach. A case report. AB - Amelogenesis imperfecta is a hereditary developmental disorder of the dental enamel, in both primary and permanent dentition. The main clinical characteristics are extensive loss of tooth tissue, poor esthetics, and tooth sensitivity. Transmission of the gene takes place by either autosomal, dominant X linked, or recessive modes. This clinical report describes a treatment sequence based on a multidisciplinary approach. A 21-year-old girl with hypoplastic amelogenesis imperfecta was referred to the Ege University School of Dentistry clinic. She was concerned about the poor appearance and sensitivity of her teeth. The patient presented with an anterior open bite, although orthodontic treatment had been completed previously. Periodontal gingivectomy of her posterior teeth followed by endodontic treatment where indicated was proposed. The prosthodontic treatment consisted of metal ceramic fixed partial dentures of precious alloy. At the end of treatment, function and esthetics were improved to a level acceptable to both the patient and the dental team. PMID- 14765636 TI - In vivo caries formation in enamel following argon laser irradiation and combined fluoride and argon laser treatment: a clinical pilot study. AB - OBJECTIVE: This in vivo pilot study investigated the role of argon laser irradiation and combined fluoride and argon laser treatment in accelerated natural caries development in sound enamel surfaces beneath plaque-retentive orthodontic bands. METHOD AND MATERIALS: Five patients (3 female, 2 male, ages 19 to 28 years) requiring tooth extraction prior to orthodontic treatment, participated in the study. Buccal surfaces were treated with either: (1) argon laser (250 mW for 10 seconds, ARGO-MOD); (2) topical fluoride (0.5% fluoride ion, Thera-Flur-N) followed by argon lasing; or (3) no treatment (control). Orthodontic bands with plaque-retentive slots on buccal surfaces were placed on the teeth slated for extraction (n = 14). Following a minimum of 5 weeks of intraoral exposure, the teeth were extracted for laboratory analysis. The teeth underwent serial longitudinal sectioning (12 sections per tooth). The sections were imbibed in water, and lesion depths were determined with each section, using polarized light microscopy. Comparisons were made among treatment groups (analysis of variance, Duncan's multiple range test for paired samples). RESULTS: Mean lesion depths were: 261 +/- 24 microm for the no treatment control group (n = 84 sections); 147 +/- 18 microm for the argon laser group (n = 24 sections); and 99 +/- 12 microm for the fluoride and argon laser group (n = 60 sections). Both the argon laser (44%) and the fluoride and argon laser groups (62%) had significant lesion depth reductions compared to controls. The addition of fluoride treatment prior to argon lasing resulted in a 32% reduction in lesion depth compared to argon laser treatment alone. CONCLUSIONS: Within this clinical pilot study, in vivo natural caries formation was affected significantly by a single exposure to low fluence argon laser irradiation. Topical fluoride treatment in combination with argon lasing provided an even greater degree of resistance against in vivo enamel caries development. A simple technique for reducing the caries susceptibility of enamel may be a clinical reality. PMID- 14765637 TI - Bonding to worn or fractured incisal edges: shear bond strength of new adhesive systems. AB - OBJECTIVES: Resin composites can be proposed for the restoration of incisal edges. If a progressive wear or an old fracture is responsible for the loss of the incisal edge, the bonding substrate is likely to be sclerotic dentin, surrounded by enamel. The purpose of the study was to measure under laboratory conditions, the shear bond strength of three adhesive systems to the enamel and dentin exposed on the edge of incisors as a result of a long-time wear or an old fracture. METHOD AND MATERIALS: Thirty extracted human incisors were selected. The teeth had to exhibit some exposed dentin on their edges, a result of old fractures or wear. The 30 specimens were randomly divided into three groups. In each group, a different adhesive system was tested for the ability to efficiently bond a resin composite restoration to the dental substrate. A three-step system (Scotchbond Multipurpose Plus), a one-bottle adhesive (Scotchbond 1), and a self etching adhesive (Prompt-L-Pop) were compared. The shear bond strength of the restored samples was measured under a loading machine. The pattern of failure that each sample underwent as a result of loading was assessed under a stereomicroscope. RESULTS: The highest values of bond strength were yielded by Scotchbond Multipurpose Plus (28.14 MPa), followed by Scotchbond 1 (16.15 MPa), and Prompt-L-Pop (9.26 MPa). These differences were statistically significant (P < .05). The most frequent pattern of failure was the combined adhesive-cohesive in resin fracture. CONCLUSION: Adhesive systems involving phosphoric acid etching of the substrate were more dependable than a self-etch adhesive when bonding a resin composite restoration to worn or fractured incisal edges. PMID- 14765639 TI - Transurgical restoration in the absence of attached gingiva. A case report. AB - A 45-year-old woman needing a Class V restoration in a mandibular molar without attached gingiva is presented. In an interdisciplinary intervention, transurgical restoration was accomplished, including flap surgery with a slight ostectomy and restoration with a bonding system and a resin composite. A 1-year clinical evaluation showed satisfactory marginal adaptation and restoration contour associated to gingival tissue health. PMID- 14765638 TI - In vitro microleakage of packable composites in Class II restorations. AB - OBJECTIVES: To evaluate the microleakage in Class II resin restorations at different margins and the polymerization shrinkage of the composites used. METHOD AND MATERIALS: Four standardized Class II (3 x 5 x 2 mm) cavities were prepared in 32 teeth. The sample had the gingival margin either 1 mm below or above the cementoenamel junction. Teeth were divided and restored according to the following protocols: (1) Single Bond + P60; (2) Prime & Bond NT + Surefil; (3) Bond-1 + Alert; and (4) Prime & Bond 2.1 + TPH. After 7 days, the specimens were thermocycled (500 cycles between 5 to 55 degrees C with a 15-second dwell time), immersed in 0.5% methylene blue for 24 hours, sectioned, and evaluated (both surfaces) at the gingival margin by two examiners, using a 0 to 3 marginal infiltration score system. The polymerization shrinkage of the composites (n = 6) was evaluated by the disk deflective method. Microleakage data was evaluated by nonparametric repeated measures ANOVA. The percentage of polymerization shrinkage was evaluated by ANOVA and Tukey's test. RESULTS: Only protocol 3 showed a significant difference between enamel and cementum margin. No difference was detected among the protocols in the enamel margin. Only protocol 1 provided a good seal in the cementum margin. All packable composites showed lower polymerization shrinkage than the hybrid tested. CONCLUSION: All protocols are able to prevent dye penetration in enamel margins; however, protocol 1 is preferable to reduce the microleakage in the cementum margin. The packable composites showed lower polymerization shrinkage compared to the hybrid resin. PMID- 14765640 TI - Refractory major aphthous stomatitis managed with systemic immunosuppressants: a case report. AB - This article presents an unusual case of major recurrent aphthous stomatitis that was refractory to multiple topical and systemic immunosuppressive therapies. Ultimately, thalidomide was selected despite its well-recognized adverse potential, and was successful in producing remission of ulcers. Strict clinical protocols were followed for this therapy in collaboration with numerous medical providers. This case illustrates the ability of multiple oral health and medical providers to collaborate in the diagnosis, management, and follow-up of a patient with an oral vesiculoerosive disease. PMID- 14765641 TI - An atypical Hallermann-Streiff syndrome. Focus on dental care and differential diagnosis. AB - The Hallermann-Streiff syndrome (HSS) is a rare congenital disorder characterized by dyscephaly, birdlike facies, hypoplastic mandible, congenital cataracts, microphthalmia, hypotrichosis, skin atrophy, proportionate short stature, and dental anomalies. A case of a 29-year-old man with atypical HSS with neither cataracts, hair and skin alterations, nor short stature is reported, with special consideration to oral findings and dental management. Dental extractions, scaling, restorations, and endodontics were performed under local anesthesia. Later, orthodontic rehabilitation with fixed brackets was carried out. Finally, a removable partial denture for the maxillary arch was designed using transparent acrylic, and this also served as a retention splint. Young patients with HSS and other similar syndromes must be involved in personalized oral health prevention programs as early as possible. Despite numerous systemic anomalies, some of these patients may undergo conventional dental procedures under local anesthesia in the dental office. PMID- 14765642 TI - In vitro effect of cavity disinfectants on the bond strength of dentin bonding systems. AB - OBJECTIVE: To evaluate the effect of two cavity disinfectants, a 2% chlorhexidine and a 1% benzalkonium chloride solution, on the shear and tensile bond strengths of dentin bonding systems to dentin. METHOD AND MATERIALS: Superficial dentin was exposed from 84 freshly extracted human third molars. The teeth were randomly assigned to two main groups according to the bonding agent used, either One Step or Optibond Solo. Each dentin bonding system had six test groups (three for shear, three for tensile testing), and each test group had a control and two cavity disinfectant groups. In the control groups, dentin bonding systems were applied after etching the dentin, whereas in the cavity disinfectant groups, dentin was conditioned and treated for 20 seconds with the disinfectants before applying the dentin bonding systems. A hybrid resin composite then was applied to all treated samples. After storage in distilled water at 37 degrees C for 24 hours, shear and tensile tests were performed. Data were analyzed using Kruskall Wallis and Mann-Whitney U tests. RESULTS: The mean shear and tensile bond strengths of One Step and Optibond Solo were not significantly different from each other, and the cavity disinfectants also had no significant effects on shear and tensile bond strength values versus the controls. CONCLUSION: The results indicate that the use of 2% chlorhexidine and 1% benzalkonium chloride solutions as cavity disinfectants after etching the dentin did not affect the shear and tensile bond strengths of One Step and Optibond Solo. PMID- 14765643 TI - Evaluation of infection-control practices by orthodontists in Turkey. AB - The aim of this study was to investigate the cross-infection control procedures used by Turkish orthodontists. A questionnaire was mailed to 268 orthodontists to determine the attitude toward infection-control procedures. Responses were received from 194 orthodontists, and the reply data were analyzed. Of the 194 orthodontists who completed and returned the questionnaire, 77.3% had been vaccinated for hepatitis B, and 24.2% always wear a mask when treating their patients. While 48.5% of the respondent orthodontists always wear gloves, 4.1% never wear gloves. PMID- 14765644 TI - Role of nutrition in the dental practice. AB - The current climate of health care delivery emphasizes comprehensive care, with an increased focus on early detection of health risk, disease prevention, and health promotion with referral to specialty providers as needed. The integration of nutrition screening, basic diet education/counseling, and referral in dental practice fits with this climate. Shifting patterns in disease epidemiology, the advent of new therapies and drugs which impact nutrition and oral health status, and greater recognition of the relationship between nutrition and oral health provides dietetics and dental professionals with increasing opportunities to create a new oral health care paradigm. This integration of oral and nutrition health care will yield improved oral, nutritional, and systemic health status. PMID- 14765645 TI - Biovigilance. PMID- 14765646 TI - Life table characteristics of Anopheles arabiensis (Diptera: Culicidae) under simulated seasonal conditions. AB - Life tables provide a summary of mortality and reproductive schedules and can help explain why certain species proliferate in particular environments. Life table characteristics of Alopheles arabiensis, under a range of simulated seasonal temperature were determined. F1 progeny of field-collected material were observed at four fluctuating temperatures with means of 17.9, 23.2, 26.1, and 21.4 degrees C, representing winter, spring, summer and autumn temperature profiles. Gonotrophic cycles, egg hatchability rates, mean male and female lifespans, reproductive rate, the intrinsic rate of increase, and generation times were calculated. Immature development was fastest in summer (11 d) compared with winter (32 d), resulting in less robust adults. Mean female lifespans were greater than those of the males and also increased with decreasing temperatures. From the age-specific survivorship curve and the gonotrophic cycles, females were found to lay large numbers of eggs especially during spring and summser. Under simulated winter conditions mosquitoes did not oviposit in the laboratory. It was also determined that summer reared mosquitoes are capable of giving just one infected bite while autumn reared mosquitoes can give two infected bites, increasing their transmission potential. Thus life table data go some way toward explaining late season transmission of malaria in South Africa. PMID- 14765647 TI - Vertical transmission of West Nile Virus by three California Culex (Diptera: Culicidae) species. AB - Three California Culex species previously identified as efficient laboratory vectors of West Nile (WN) virus were tested for their capability to vertically transmit WN virus. Wild-caught Culex pipiens pipiens L., Culex pipiens quinquefasciatus Say, and two populations of Culex tarsalis Coquillett females were inoculated intrathoracically with 10(2.7 +/- 0.1) plaque-forming units of WN virus. F1 progeny were reared at 18 degrees C and subsequently tested as adults for infectious virus on Vero cell culture. Virus was not detected in 197 pools comprising 4,884 Cx. p. pipiens. The minimum filial infection rate (MFIR) for Cx. p. quinquefasciatus was approximately 3.0/1,000 for 665 progeny tested in 28 pools. There was no virus detected in 102 pools of 2,453 progeny from Cx. tarsalis collected in Riverside County. The MFIR for Cx. tarsalis collected in Yolo County was approximately 6.9/1,000 for 2,165 progeny tested in 86 pools. Mosquito progeny infected vertically during the fall could potentially serve as a mechanism for WN virus to overwinter and initiate horizontal transmission the following spring. PMID- 14765648 TI - Spiracular indices in Anopheles stephensi: a taxonomic tool to identify ecological variants. AB - Thoracic spiracle length and its index was examined for their ability to discriminate two ecological variants, type form and mysorensis, of Anopheles stephensi in the adult stage. The type form is exclusively domestic in all seasons, whereas the mysorensis variant occupies the outdoor niche during monsoon and postmonsoon seasons, with spillover into domestic sites during summer ecological stress periods. A statistically significant co-relation was established between the ridge count of the egg and two adult measurements, the thoracic spiracle length, and the spiracular index. In An. stephensi type form, average spiracle length was 0.11-0.12 mm and average spiracular index was 8.09 9.23, whereas in mysorensis, the corresponding figures were 0.09-0.10 mm and 6.82 7.60. These parameters showed consistent variations in population of mosquitoes that emerged during monsoon and summer season. The thoracic lengths in both variants remained constant, and only spiracular lengths showed fluctuations in three seasonal populations. These measures provide discrimination of adult variants--identifications that are essential in malaria control programs. PMID- 14765649 TI - Multivariate morphometric discrimination among three species of Lutzomyia subgenus Micropygomyia (Diptera: Psychodidae). AB - Canonical discriminant analysis accurately separated males of Lutzomyia yencanensis (n = 36) (Ortiz), Lutzomyia cayennensis cayennensis (n = 24) (Floch and Abonnenc) and Lutzomyia micropyga (n = 28) (Mangabeira). Only seven (length of head, interocular distance, wing vein delta, genital filaments, coxite, paramere, and lateral lobe) of 20 possible morphometric measurements were required for differentiation, with a highly significant canonical correlation. The discriminant equations provided a simple, fast, and widely available statistical tool to identify accurately unknown sand fly specimens as L. yencanensis, L. c. cayennensis, or L. micropyga. PMID- 14765650 TI - Abdominal pericardial sinus: a neurohemal site in the tsetse and other cyclorraphan flies. AB - An ultrastructural study of the heart of the tsetse fly, Glossina morsitans, and of several other species of cyclorraphan flies revealed that the ventral region of the heart of adult flies is supported by a muscular septum not present in the larval stage. The pericardial septum of the adult heart is composed laterally of alary muscles and a central longitudinal muscle that extends the length of the abdominal aorta, whereas the larval heart is supported ventrally only by alary muscles and strands of connective tissue. Thus, unlike the larval stage, and the heart of other insects, the pericardial septum of adult cyclorraphan flies contains a central band of longitudinal muscle, that along with the alary muscle, forms a large pericardial sinus lying between the septum and the heart. Neurosecretory nerves arising from the lateral nerves of the thoracicoabdominal ganglion extend dorsad to the pericardial septum, where they form neuromuscular junctions on the muscle fibers of the pericardial septum or traverse the septum terminating in the pericardial sinus, thereby creating one of the largest neurohemal organs in these flies. In the tsetse fly, some of the neurosecretory fibers also extend between the muscle fibers of the myocardium, and release their material into the lumen of the heart. PMID- 14765651 TI - Argas (Persicargas) keiransi n. sp. (Acari: Argasidae), a parasite of the Chimango, Milvago c. chimango (Aves: Falconiformes) in Chile. AB - The larva of Argas (Persicargas) keiransi Estrada-Pena, Venzal & Gonzalez-Acuna n. sp. is described from specimens collected on the neck of a chimango, Milvago c. chimango (Aves: Falconiformes) in the Chillan, Chile, in the Sub-Antarctic biogeographical region. The larva of the new species shares the tarsus I setal formula with A. (P.) giganteus, these being the only two Persicargas species with three pairs of ventral setae plus both av4 and pv4 setae. However, it is unique in having a dorsal plate "V" or "U" shaped, with the anterior end open, without the typical reticulated pattern present in the remaining species of the genus. PMID- 14765652 TI - Morphological and populational characteristics of hemocytes of Ornithodoros moubata nymphs during the ecdysial phase. AB - The ultrastructure and characteristics of hemocytes of argasid tick species, Ornithodoros moubata, during the ecdysdial phase are herein presented. Hemocyte classes/populations characterized based on their affinity with Giemsa stain and ultrastructural differences comprised the prohemocytes, nongranular cells (Nc), eosinophilic granular cells (Ec), basophilic granular cells (Bc), and unidentified cells. Significant changes/shift in the ratio of hemocyte classes/population was apparent in ticks before and after the ecdysial phase. The granule-scant basophilic granular cells (sBc) constituted the most abundant hemocyte population in the ecdysial phase. Nymphs in ecdysis showed increases in Nc and sBc and decrease in Ec, a phenomenon that was reversed in unengorged nymphs and adults ticks. The significant increase in total Bc population in ecdysis relative to nonengorged ticks clearly point to blastogenesis of Bc taking place during the ecdysial phase and Bc's important role in the process of tissue remodeling. PMID- 14765653 TI - Occurrence and distribution of Anopheles (Diptera: Culicidae) larval habitats on land cover change sites in urban Kisumu and urban Malindi, Kenya. AB - A multitemporal, land use land cover (LULC) classification dataset incorporating distributions of mosquito larval habitats was produced in ERDAS Imagine using the combined images from the Multispectral Thermal Imager (MTI) at 5 m spatial resolution from 2001 with Thematic Mapper-classification data at 28.5 m spatial resolution from 1987 and 1989 for Kisumu and Malindi, Kenya. Total LULC change for Kisumu over 14 yr was 30.2%. Total LULC change for Malindi over 12 yr was 30.6%. Of those areas in which change was detected, the LULC change for Kisumu was 72.5% for nonurban to urban, 21.7% urban to nonurban, 0.4% urban to water, 4.5% water to urban, and 0.9% water to nonurban. The proportion of LULC change for Malindi was 93.5% for nonurban to urban, 5.9% urban to nonurban, 0.2% urban to water, 0.3% nonurban to water, and 0.1% water to urban. A grid (270 m x 270 m cells) was overlaid over the maps stratifying grid cells based on drainage and planning. Of 84 aquatic habitats in Kisumu, 32.1% were located in LULC change sites and 67.9% were located in LULC nonchange sites. Of 170 aquatic habitats in Malindi, 26.5% were located in LULC change sites and 73.5% were located in LULC nonchange sites. The most abundant LULC change per strata with anopheline habitats was unplanned and poorly drained. Ditches and puddles in Kisumu and car tracks in Malindi displayed the highest number of anopheline larval habitats for all LULC change sites. The proportion of site positive aquatic habitats for anopheline larvae was higher in LULC change sites than for LULC nonchange sites for Kisumu. This evidence suggests LULC change can influence anopheline larval habitat distribution. PMID- 14765654 TI - Convergent habitat segregation of Aedes aegypti and Aedes albopictus (Diptera: Culicidae) in southeastern Brazil and Florida. AB - During the rainy season of 2001, the incidence of the dengue vectors Aedes aegypti and Ae. albopictus was examined in different habitats of two cities (Rio de Janeiro and Nova Iguacu) in Rio de Janeiro State, Brazil, and in two cities (Palm Beach and Boca Raton) in Florida. Oviposition trap collections were performed in urban, suburban, and rural habitats in both areas. Our hypothesis that the abundances and frequencies of occurrence of Ae. aegypti and Ae albopictus are affected in opposite ways by increasing urbanization was only partially supported. City, habitat, and their interaction significantly affected the abundance of both species. Cities with high abundance of Ae. aegypti also had a high abundance of Ae. albopictus. The two species were most abundant in the cities of Rio de Janeiro state and the lowest in Boca Raton. Habitat had a significant but opposite effect on the abundances of Ae. aegypti and Ae. albopictus. In general, Ae. aegypti was most prevalent in highly urbanized areas and Ae. albopictus in rural, suburban, and vegetated urban areas in Rio de Janeiro state and Florida. However, abundances of the two species were similar in most suburban areas. Analyses of frequencies of occurrence showed an unexpected high level of co-occurrence of both species in the same oviposition trap. Despite the different geographical origins of Ae. albopictus in Brazil and the United States, the habitats used by this recent invader are remarkably similar in the two countries. PMID- 14765655 TI - An automated feeding apparatus for in vitro maintenance of the human head louse, Pediculus capitis (Anoplura: Pediculidae). AB - An automated feeding apparatus was developed to maintain the human head louse (Pediculus capitis DeGeer) in vitro. With the use of valves and timers, banked human blood and saline from refrigerated reservoirs were pumped into and flushed out of the system every 7 d. During this rotational interval, bloodmeals were provided to head lice continuously and ad libitum through a stretched Nescofilm silicone sandwich membrane. Compared with our previous in vitro human head louse rearing apparatus, greater numbers of lice could be fed simultaneously with minimal human monitoring. Development of second to third instars and third instars to adults was significantly faster when lice were reared in vivo than on either of the in vitro rearing systems; there was no significant difference in the duration of the first instar. Although fecundity and hatch rates were significantly higher for female lice reared in vivo, similar trends have been observed for other membrane-fed arthropods. Body lice (Pediculus humanus L.) and bed bugs (Cimex lectularius [L.]) also completed most of their life cycle on this apparatus. Our automated mass-rearing system has broad applications for maintaining fluid-sucking ectoparasites and will facilitate various toxicological, behavioral, and disease-transmission investigations. PMID- 14765656 TI - Habitats, dispersion and invasion of sylvatic Triatoma dimidiata (Hemiptera: Reduviidae: Triatominae) in Peten, Guatemala. AB - Dispersion and invasion capacity of sylvatic populations of Triatoma dimidiata (Latreille) were investigated during 14 mo by means of experimental chicken coops installed in a nature reserve within the Maya Biosphere, Peten, Guatemala. In addition, palm trees, underground archeological holes (chultunes) and piles of limestones within the forest were inspected as potential sylvatic habitats of T. dimidiata. From the three types of sylvatic habitats we inspected, all served as shelter and breeding sites for T dimidiata. The natural infection of these bugs (n = 72) with Trypanosoma cruzi (Chagas) was high (25%) and represent a risk for humans who colonize the forest. T. dimidiata quickly invaded the experimental chicken coops installed in the primary forest, albeit at very low densities. However, only one adult bug was encountered in the chicken coops installed in a secondary forest. Dispersal of adult T. dimidiata was most apparent at the end of the dry season. Overall, our results indicate a potential risk for invasion by sylvatic T. dimidiata of domestic environments in this area, with a risk of T. cruzi transmission to humans. We suggest that a system of community-based surveillance should be developed to detect new infestations and organize prompt treatment of any new cases of acute Chagas disease that may result. PMID- 14765657 TI - Biting rates and developmental substrates for biting midges (Diptera: Ceratopogonidae) in Iquitos, Peru. AB - Biting midges (Diptera: Ceratopogonidae) were collected at 16 periurban and rural sites around Iquitos, Peru, between 17 October 1996 and 26 May 1997. Culicoides paraensis (Goeldi), the principal vector of Oropouche virus, was the most commonly collected species (9,086 flies) with Culicoides insinuatus Wirth & Blanton second (7,229 flies). Although both species were collected at all sampling sites (linear (distance surveyed approximately 25 km), C. paraensis dominated at northern collection sites (> 90%), whereas C. insinuatus prevailed at southern collection sites (> 60%). C. paraensis were collected from human sentinels at a constant rate throughout daylight hours, at similar rates during wet and dry months, and regardless of rainfall. Larval developmental substrates for C. paraensis included decaying platano (Musa x paradisiaca L. [Musaceae]) stems, stumps, flowers, fruits, and debris beneath platano trees as well as from soil beneath a fruiting mamay (Syzygium malaccense Merr. & Perry [Myrtaceae] ) tree and organic-rich mud along a lake shoreline. C. insinuatus adults likewise emerged from decaying platano and organic-rich mud along a lake shoreline, but also from debris accumulated in the axils of aguaje (Mauritia flexuosa L. [Palmae]) fronds and decaying citrus fruit. Despite high numbers of biting adults near putative substrates, adults of neither species emerged from other decomposing plant material, soil, phytotelmata, or artificial containers. Because both species of biting midges emerged in high numbers from all parts of platano (ubiquitous in Iquitos), it will be challenging to control them through sanitation. PMID- 14765658 TI - Ectoparasite associations of bats from central Pennsylvania. AB - Between April and October 1997, 689 bats representing seven species were captured at Pennsylvania's Canoe Creek State Park. Each bat was sampled for ectoparasitic arthropods, and four species were collected from 13.2% of the host individuals. Ectoparasites include the bat flea Myodopsylla insignis (Rothschild), the wing mite Spinturnix americanus (Banks), the bed bug Cimex adjunctus Barber, and the soft tick Ornithodoros kelleyi Cooley & Kohls. Prevalence, relative density, and mean intensity were calculated for ectoparasites of Myotis lucifugus (Le Conte), which harbored all four ectoparasite species and was the most commonly captured host. Patterns of ectoparasite associations were examined with respect to host sex and habitat (roost characteristics). Female M. lucifugus hosted higher densities of ectoparasites than did males. Moreover, relative densities of ectoparasites from M. lucifugus were dependent on the proximate roost; hosts captured near Bat Church were more heavily parasitized than those captured near Hartman Mine. Two other bat species were infested with at least one ectoparasite, but sample sizes were too small to analyze statistically. These bat species included Myotis septentrionalis (Trouessart), harboring M. insignis, S. americanus, and O. kelleyi, and Eptesicus fuscus (Beauvois), which harbored M. insignis and O. kelleyi. PMID- 14765659 TI - Factors influencing the outcome of mark-release-recapture studies with Culex tarsalis (Diptera: Culicidae). AB - Three potentially important aspects of mark-release-recapture experimentation were addressed: 1) source of mosquitoes for release, 2) time of release, and 3) weather during recapture. Culex tarsalis Coquillett mosquitoes collected as adult host-seeking females from dry ice-baited traps (CO2 traps) operated within the study area (local) were recaptured more frequently than females collected from traps operated outside the study area (foreign) or reared from field-collected immatures (reared). These results supported published studies on Anopheles and Ochlerotatus that indicated mosquitoes may "memorize" flight paths within their environment. Releasing gravid females provided a potentially useful replacement for reared females, because these gravids oviposited at wetlands and then dispersed to upland traps. Releasing local, foreign, or reared mosquitoes just after sunrise or just before sunset did not alter recapture success or the distance dispersed. Elevated wind speeds inhibited dispersal from protected microhabitats with citrus orchards and resulted in most recaptures being found at the leeward portion of the orchard. PMID- 14765660 TI - The impact of selective oviposition and infection with Plagiorchis elegans on Aedes aegypti pre-imago population dynamics at optimal food availability. AB - Progressive changes in the attraction of waters harboring pre-imago populations of Aedes aegypti exposed to different levels of the entomopathogenic digenean Plagiorchis elegans to ovipositing conspecific females were assessed under conditions of optimal food availability. The impact of ovipositional preference and parasitic infection on population structure and development was investigated. Probabilities that larvae progress from one stage to the next or die within 24 h were calculated for all life stages. Exposure to P. elegans cercariae did not significantly affect the attractiveness of larval-holding waters. Ovipositional preference increased significantly with growing bio-mass of the larval population, with the event of pupation and, in some cases, with late instar mortality. Exposure to various levels of the parasite significantly increased mortality of all instars, but most of the damage caused by the parasite occurred in the form of increased pupal mortality and decreased adult emergence. Exposure to the parasite significantly reduced the number of adults produced yet did not impair larval development. Thus, larval recruitment into environments containing P. elegans remains high, the structure of larval populations remains relatively normal, but few adults are produced. PMID- 14765661 TI - Role of bacteria in mediating the oviposition responses of Aedes albopictus (Diptera: Culicidae). AB - The responses of Aedes albopictus to sources of oviposition attractants and stimulants were evaluated with a behavioral bioassay in which females attracted to odorants emanating from water were trapped on screens coated with an adhesive. Gravid mosquitoes were attracted to volatiles from larval-rearing water and soil contaminated cotton towels. Bacteria were isolated from these substrates and from an organic infusion made with oak leaves. Through fatty acid-methyl ester analyses, six bacterial isolates from larval-rearing water, two isolates from soil-contaminated cotton towels, and three isolates from oak leaf infusion were identified to species. The response of gravid mosquitoes to these isolates was also evaluated in behavioral bioassays. Water containing Psychrobacter immobilis (from larval-rearing water), Sphingobacterium multivorum (from soil-contaminated cotton towels), and an undetermined Bacillus species (from oak leaf infusion) elicited significantly higher oviposition than control water without bacteria. Only volatiles collected from larval rearing water elicited significant electroantennogram responses in females. PMID- 14765662 TI - Chemical composition of some components of the arrestment pheromone of the black legged tick, Ixodes scapularis (Acari: Ixodidae) and their use in tick control. AB - Chemical analysis (high-performance liquid chromatography) and bioassay demonstrated the presence of compounds that seem to be components of the Ixodes scapularis arrestment pheromone. Only two purines, guanine and xanthine, were found in acidified saline extracts made from cast skins after molting of fed nymphs, fed larvae, and fecal/excretory exudates deposited by unfed adults on substrates in their environment. The ratio of guanine to xanthine was 10.6:1 in an extract from the nymphal skins versus 0.95:1 in an extract from the larval skins. Guanine, xanthine, and traces of a third purine, tentatively identified as 8-azaguanine, were found in extracts made from filter paper strips or washings from glass vials contaminated with tick feces and excreta left by unfed adults. 8 azaguanine may be a product of microbial degradation of the other purines rather than a natural product from the ticks. Low concentrations of ammonia also were detected in saline extracts of excreta from feeding ticks. Hematin also was found in NH4OH extracts of the black fecal/excretory exudates deposited by the unfed ticks. Hematin was tentatively identified by comparison of spectra with that of the authentic standard. Bioassays demonstrated a strong positive arrestment response to cast skins found to contain a mixture of guanine and xanthine and to black fecal/excretory exudates containing guanine, xanthine, the putative 8 azaguanine, and hematin. A Noldus video tracking system using a CCD video camera and Ethovision Pro tracking software showed statistically significant increases in the frequency of visits to the treated zone versus the control. Ticks were significantly more likely to assemble in response to the tick exudates within as little as 3 h compared with the controls. Previous bioassay studies also showed strong positive responses to guanine, xanthine, other purines, and hematin. Comparisons with the arrestment pheromones of other tick species are described. The inclusion of the pheromone components in a permethrin-impregnated oily matrix, Last Call, increased the lethal activity of the product to 95% compared with only 65% in the formulation with permethrin alone. More detailed knowledge of I. scapularis arrestment pheromone may be useful for improving the efficacy of this tick-killing technology even further. PMID- 14765663 TI - A component of maize pollen that stimulates larval mosquitoes (Diptera: Culicidae) to feed and increases toxicity of microbial larvicides. AB - To explain how larval Anopheles arabiensis Patton feed effectively in the turbid water in which they frequently develop, we determined whether an extractable component of maize, Zea mays L., pollen enhances feeding by these mosquitoes. Maturing maize produces a copious amount of wind-borne pollen that is nutritious enough and produced over a sufficient period to support the development of at least one generation of anopheline mosquitoes. Larval An. arabiensis readily ingest the contents of maize pollen or the intact pollen grains themselves. An aqueous extract of maize pollen markedly accelerates the rate at which larval An. arabiensis ingest inert particles and strongly enhances the effectiveness of Bti against larval An. arabiensis. We conclude that the ability of larval anopheline mosquitoes to feed on maize pollen in turbid water is enhanced by the release from these pollen grains of a water-soluble phagostimulatory component (or components), which may be used to increase ingestion of microbial entomotoxins. PMID- 14765664 TI - Genetic differentiation of Anopheles claviger s.s. in Europe. AB - Two groups of Anopheles claviger sensu stricto Meigen (Diptera: Culicidae) were recently found in France, representing unclear genetic entities. To better understand this situation, sampling was extended to 13 European countries, and 47 samples were analyzed by investigating the polymorphism of 11 autosomal and 1 sex linked allozyme loci. Genetic differentiation, as measured by F(st), between these two groups was confirmed, with no isolation by distance within each group. Among the twelve loci studied, none had diagnostic alleles. Group I is mainly located in western Europe (UK and south-west of France), and Group II covers eastern France and eastern and northern Europe. Intermediate populations, sampled at the overlapping range between them, do not display a heterozygote deficit, suggesting that these two groups are probably not genetically isolated. The origin of each group and its biological significance is discussed within the context of differentiation in refugia during recent glaciations. PMID- 14765665 TI - Seasonal abundance and blood feeding activity of Anopheles minimus Theobald (Diptera: Culicidae) in Thailand. AB - Anopheline mosquito larvae and adults were sampled at Ban Pu Teuy, Tri-Yok District, Kanchanaburi Province, western Thailand, from January 2000 to December 2001. Over the period of 2 yr, Anopheles minimus sensu lato was the most commonly collected species, followed by Anopheles swadiwongporni and Anopheles dirus sensu lato; all three species are important vectors of malaria in Thailand. Attempted blood feeding by An. minimus occurred throughout the night, with two distinct feeding peaks: strong activity immediately after sunset (1800-2100 hours), followed by a second, less pronounced, rise before sunrise (0300-0600 hours). Anopheles minimus were more abundant during the wet season compared with the dry and hot seasons, although nocturnal adult feeding patterns were similar. Anopheles minimus fed readily on humans inside and outside of houses, showing a slight preference for exophagy. The human-biting peak of An. minimus in our study area differed from other localities sampled in Thailand, indicating the possible existence of site-specific populations of An. minimus exhibiting different host seeking behavior. These results underscore the importance of conducting site specific studies to accurately determine vector larval habitats and adult activity patterns and linking their importance in malaria transmission in a given area. PMID- 14765666 TI - Temporal and geographic genetic variation in Culex pipiens quinquefasciatus (Diptera: Culicidae) from Florida. AB - Culex (Culex) pipiens quinquefasciatus Say field population from Vero Beach, FL, sampled monthly over a period of 8 mo, a colony sample, and six geographic samples were analyzed for genetic variation at 12 enzymes (10 "neutral" gene enzymes with 11 putative loci and two "complex" gene enzymes) by using polyacrylamide gel electrophoresis. The analysis of the 11 putative loci in both temporal and geographic samples showed that the four loci (Gpi, Hk, Mdhp-2, and Pgm) diagnostic of Cx. p. quinquefasciatus in the southern United States are present in similar frequencies in Florida samples. The Cx. p. quinquefasciatus colony sample showed significantly lower genetic variation than the temporal field samples, measured by mean number of alleles per locus (colony 1.2 +/- 0.1 versus field 1.44 +/- 0.03), percentage of polymorphic loci (colony 18.2% versus field 28.4%), mean observed heterozygosity (H(o) = colony 0.027 +/- 0.02 versus field 0.09 +/- 0.01), and mean Hardy-Weinberg expected heterozygosity (H(e) = colony 0.025 +/- 0.02 versus field 0.085 +/- 0.01). Three of the 11 loci (Acoh, Pgd, and Pgm) from the Vero Beach field samples showed bimodal patterns in their frequencies of the most common allele during peak density of the population. The low value of F(st) of 0.058 indicated minimum population substructuring among the temporal samples. Genetic variability values between geographic samples from the Florida panhandle and south Florida were not significant. Gene flow estimates based on F(ST), = 0.05, indicating low levels of gene flow among the geographic samples of Cx. p. quinquefasciatus. The average Nei's and modified Rogers' genetic distances among the six populations were 0.005 +/- 0.001 and 0.077 +/- 0.007, respectively. The cluster analysis did not suggest geographic clustering. The analysis of the "complex" gene enzymes in both temporal and geographic samples of Cx. p. quinquefasciatus from Florida showed the presence of two highly amplified esterases (Estbeta1 and Estalpha2?Estbeta2), indicating resistance to organophosphate insecticides and highly amplified Aldox enzyme (an enzyme that indicates resistance to at least one insecticide and a herbicide). Comparison of our results with previous studies on Cx. p. quinquefasciatus populations in the United States indicates that the genetic characteristics of the Florida populations of Cx. p. quinquefasciatus are very similar to populations from areas where ecological conditions are very different. PMID- 14765667 TI - Cloning and sequencing of putative acetylcholinesterase cDNAs from the American dog tick, Dermacentor variabilis, and the brown dog tick, Rhipicephalus sanguineus (Acari: Ixodidae). AB - Two putative cDNAs of acetylcholinesterase (AChE), one from Dermacentor variabilis, and the other from Rhipicephalus sanguineus, were amplified and sequenced. The deduced amino acid sequences have high amino acid identities (between 70 and 94%) to known tick AChE sequences deposited in GenBank. Furthermore, these two AChEs also possess common features in their primary AChE structure such as catalytic active sites. A 2,220-bp contiguous sequence, containing a 1,791-bp open reading frame encoding an AChE precursor with 596 amino acid residues, was obtained from D. variabilis. The deduced proteins of R. sanguineus are different in size by 6 amino acids because of alternative splicing at the 5' end. A gene tree deduced from phylogenetic analysis indicates that there are at least three lineages of AChE in arthropods. PMID- 14765668 TI - Autogeny in Ochlerotatus vigilax (Diptera: Culicidae) from southeast Queensland, Australia. AB - Field and laboratory investigations were undertaken to determine the level of expression of autogeny in the mosquito Ochlerotatus vigilax (Skuse) from southeast Queensland, Australia, and whether there was evidence of seasonal variation. At two field sites in southeast Queensland, Wellington Point and Donnybrook, autogeny rates were determined on six occasions between January 2001 and January 2002. The autogeny rate varied between 71 and 100% at Wellington Point and between 63 and 100% at Donnybrook. Autogenous fecundity ranged from 17 to 63 eggs per female at Wellington Point and from 13 to 88 eggs per female at Donnybrook. Positive relationships were found between adult body size (indicated by wing length), autogeny rate, and fecundity. A laboratory study was conducted to investigate the influence of larval nutrition and adult diet (water versus sucrose) on the expression of autogeny. The autogeny rate at a low-diet treatment was between 73 and 90% when sucrose was withheld from females and 100% when sucrose was provided. All high-diet females were autogenous. Autogenous egg development required 80 +/- 6 h from emergence at 27 degrees C. We conclude that autogeny rates are consistently high in Oc. vigilax from the southeast Queensland region. PMID- 14765669 TI - Metabolic rate in female Culex tarsalis (Diptera: Culicidae): age, size, activity, and feeding effects. AB - We used flow-through respirometry on female mosquitoes to observe individual ventilatory pattern and to measure metabolic rate at rest, during activity and after a blood-meal. At rest, young adult females of the species Culex tarsalis ventilated cyclically with an average VCO2 of 6.5 nl/min and frequency of 45 mHz. Both resting metabolic rate and body mass of females increased between emergence and 4 d, but only body mass continued to increase until 12 d. Mating status had no effect on the body mass or resting metabolic rate of the females. Both lipid and carbohydrate stores strongly increased during the first week of adult life, in particular between emergence and 4 d, in which lipid mass increased 8.3-fold and carbohydrate mass increased 3.6-fold. Hourly metabolic rate after a blood or sugar meal was measured over a 6-d period in two groups of 14 females. Sugar-fed mosquitoes remained calm during the day and had nocturnal increases in metabolic rate associated with flight activity. Blood feeding elicited a specific dynamic action lasting approximately 55 h, at the peak of which metabolic rate of the blood-fed females was twice that of the sugar-fed group. Eggs were laid on the third night. The increase in metabolic rate presumably reflected the cost of blood digestion and egg production. The females were not active during digestion, so that although their metabolic rate was increased, the overall energy expenditure of the blood-fed group was not very different from that of the sugar fed group. PMID- 14765670 TI - Efficacy of pyrethroid insecticides against domestic and peridomestic populations of Triatoma pallidipennis and Triatoma barberi (Reduviidae:Triatominae) vectors of Chagas' disease in Mexico. AB - A single village control trial for Triatoma pallidipennis and T. barberi was conducted using three synthetic pyrethroids (bifenthrin, cyfluthrin, and deltamethrin), evaluated as residual treatments in separate sectors, with complete coverage indoors and in peridomiciliary areas. Spray intervention was preceded by a preintervention entomological evaluation and household survey, followed by four postintervention evaluations at 1, 3, 6, and 12 mo of > 96% of houses. Overall preintervention adjusted infestation index was 38%, 17% of which represented intradomicile infestation. Dose verification using high-performance liquid chromatography (HPLC) demonstrated correct spray doses for all but deltamethrin treatments. There was between a 6- and 13-fold decrease in intradomicile live bug infestation for cyfluthrin- and bifenthrin-treated areas, resulting at 1 mo in 0 and 0.6% infestation, respectively. Intradomicile infestation recovered somewhat, terminating at 20 and 50% of preintervention levels at 12 mo, respectively, while peridomicile infestation recovered preintervention levels within 3-6 mo. Households with persistent live peridomiciliary infestation had 1.9 times the risk of having a persistent intradomiciliary infestation, while 80% of peridomicile infestations for both triatomine species were in houses not having a previous infestation. New or reinfestation of households did not occur consistent with a sylvan source, and unconstructed lots were not a significant source of bugs. Houses with persistent peridomicile infestation did represent a significant risk for surrounding uninfested houses by cluster analysis (P < 0.05). Along with the increased prevalence of T. cruzi infection after intervention, the data indicate that a sylvan reservoir source, probably peridomicile small rodent nests, represent the major risk factor for persistent and new infestations. PMID- 14765671 TI - Larval habitat diversity and ecology of anopheline larvae in Eritrea. AB - Studies on the spatial distribution of anopheline mosquito larvae were conducted in 302 villages over two transmission seasons in Eritrea. Additional longitudinal studies were also conducted at eight villages over a 24-mo period to determine the seasonal variation in anopheline larval densities. Eight anopheline species were identified with Anopheles arabiensis predominating in most of the habitats. Other species collected included: An. cinereus, An. pretoriensis, An. d'thali, An. funestus, An. squamosus, An. adenensis, and An. demeilloni. An. arabiensis was found in five of the six aquatic habitats found positive for anopheline larvae during the survey. Anopheles larvae were sampled predominantly from stream edges and streambed pools, with samples from this habitat type representing 91.2% (n = 9481) of the total anopheline larval collection in the spatial distribution survey. Other important anopheline habitats included rain pools, ponds, dams, swamps, and drainage channels at communal water supply points. Anopheline larvae were abundant in habitats that were shallow, slow flowing and had clear water. The presence of vegetation, intensity of shade, and permanence of aquatic habitats were not significant determinants of larval distribution and abundance. Larval density was positively correlated with water temperature. Larval abundance increased during the wet season and decreased in the dry season but the timing of peak densities was variable among habitat types and zones. Anopheline larvae were collected all year round with the dry season larval production restricted mainly to artificial aquatic habitats such as drainage channels at communal water supply points. This study provides important information on seasonal patterns of anopheline larval production and larval habitat diversity on a countrywide scale that will be useful in guiding larval control operations in Eritrea. PMID- 14765672 TI - Field evaluation of an area repellent system (Thermacell) against Phlebotomus papatasi (Diptera: Psychodidae) and Ochlerotatus caspius (Diptera: Culicidae) in Sanliurfa Province, Turkey. AB - A field evaluation of a new area repellent system, Thermacell Mosquito Repellent (TMR, cis-trans allethrin), was conducted against phlebotomine sand flies and mosquitoes in Cunpolat village, Sanliurfa Province, in southeastern Anatolia, Turkey, an area historically endemic for cutaneous leishmaniasis and high sand fly populations. The objective of this study was to determine the efficacy, duration of protection, and spatial characteristics of protection (downwind distance from point of release), of the TMR. Twelve adult volunteers (nine in the treatment and three controls) made collections from ankle to knee for 25 min every hour from 2100 to 0300 on six consecutive nights in August 2002. Treatment consisted of placing a TMR unit at the middle of the village and then placing human bait collectors at 2.3, 4.6, and 7.6 m away from the repellent unit. Results from the field tests showed highly significant protection provided by the TMR from attack by Phlebotomus papatasi (Scopoli) (P < 0.001) and Ochlerotatus caspius (Pallas) (P < 0.001) for up to 4 h postapplication. In the six nights that tests were done, a total of 949 sand flies and 1095 mosquitoes were collected from the untreated control sites. Only 86 sand flies and 83 mosquitoes were collected at all distances from the places treated with TMR. These results indicated that reduction in sand fly and mosquito biting rates in treated groups ranged from 87.5 to 97.7% (mean protection 92%) and 90.2-97.4% (mean protection 93%), respectively. The percentage reduction values were maintained above 90.0% for 6 h of the assessment period. Overall, the number of bites by the cutaneous leishmaniasis vector Ph. papatasi and also Oc. caspius was reduced > 11-fold and 13-fold, respectively, by the TMR. PMID- 14765673 TI - Contact toxicity and residual activity of different permethrin-based fabric impregnation methods for Aedes aegypti (Diptera: Culicidae), Ixodes ricinus (Acari: Ixodidae), and Lepisma saccharina (Thysanura: Lepismatidae). AB - The effectiveness and residual activities of permethrin-impregnated military battle dress uniforms were evaluated by comparing a new company-manufactured ready-to-use polymer-coating method with two "dipping methods" that are currently used to treat uniforms. Residual permethrin amounts and remaining contact toxicities on treated fabrics before and after up to 100 launderings were tested against Aedes aegypti (L.), Ixodes ricinus (L.), and Lepisma saccharina (L.). The residual amount of permethrin was considerably higher with the polymer-coating method: 280 mg a.i./m2 after 100 launderings, compared with 16 and 11 mg a.i./m2, respectively, obtained when using the two dipping methods. Hard ticks were most susceptible to the new polymer-coating method, resulting in prelaundering 100% knockdown times of 7.0 +/- 0.9 min, whereas equivalent times for the dipping methods were 7.9 +/- 0.35 min and 8.0 +/- 0.54 min, respectively. After 100 launderings, 100% knockdown of I. ricinus nymphs was reached at 15.2 +/- 1.04 min using the polymer-coating method, compared with 178.8 +/- 24.7 min and 231 +/- 53.6 min, respectively, using the dipping methods. Similar results were obtained for Ae. aegypti and L. saccharina, indicating that the polymer-coating method is more effective and efficient when compared with the dipping methods. PMID- 14765674 TI - Survey of resistance to permethrin and diazinon and the use of a multiplex polymerase chain reaction assay to detect resistance alleles in the horn fly, Haematobia irritans irritans (L.). AB - A field survey was conducted in 2001 to evaluate resistance to pyrethroid and organophosphate (OP) insecticides on horn flies, Hematobia irritans irritans (L.), from seven ranches in the state of Tamaulipas, Mexico, and from three locations in central Texas. Filter papers impregnated with either technical permethrin or diazinon were used to measure the levels of resistance to pyrethroids and OPs. A multiplex polymerase chain reaction (PCR) assay was used on individual horn flies from these field populations to detect the presence of the kdr and super-kdr alleles associated with pyrethroid resistance, and a mutated alphaE7 esterase allele associated with OP resistance. Relative to a susceptible laboratory (Kerrville) strain, horn flies from Mexico exhibited 5.1- to 28.3-fold resistance to permethrin at the LC50, and 23.8- to 136-fold resistance at the LC90. Horn flies from Texas ranches exhibited only two- to five fold resistance. All field populations of the horn fly were highly susceptible to diazinon, and no mutant alphaE7 esterase alleles were detected. The super-kdr allele was found only in a single fly from a ranch in Mexico. Results of PCR assays showed that the kdr allele was present at various frequencies in field populations of horn flies. A gender-related bias in distribution of kdr genotypes was found in horn flies from Mexico, but not in horn flies from Texas. The overall kdr allelic frequencies in horn flies from Mexico were 23.2-37.8% higher in females than in males. Regression analysis revealed a significant correlation between kdr allelic frequencies and the levels of knockdown resistance to permethrin among the horn fly populations studied. The results validate the role of the PCR-based molecular assay as a diagnostic tool in monitoring resistance to pyrethroids and also provide useful information on population genetics of horn fly resistance to pyrethroids and OPs. PMID- 14765675 TI - Enhanced vector competence of Aedes aegypti (Diptera: Culicidae) from the Torres Strait compared with mainland Australia for dengue 2 and 4 viruses. AB - Australian Aedes aegypti (L.) mosquitoes colonized from the Torres Strait and three mainland localities (Charters Towers, Townsville, and Cairns) were fed on blood suspensions containing dengue virus type 2 (DEN-2) or dengue virus type 4 (DEN-4). Variation was found in oral susceptibility to DEN-2 (59 -99% infection) and DEN-4 (28-79% infection) among Ae. aegypti assayed for virus at 8, 12, 16, or 20 d after ingestion of infected blood. Torres Strait Ae. aegypti were the most susceptible to DEN-2 and were significantly more efficient in transmission to capillary tube at 16 d (76% transmission) than mainland Ae. aegypti populations (20-28% transmission). Torres Strait Ae. aegypti were also the most susceptible to DEN-4, although transmission did not vary significantly from mainland populations at 16 d (12% compared with 0-4%) or 20 d (16% compared with 4-16%). Disseminated infection (i.e., leg infection) with either DEN-2 or DEN-4 was not an accurate predictor of transmission potential. This study demonstrates differences among Australian Ae. aegypti populations in vector competence for DEN 2 and DEN-4. Torres Strait Ae. aegypti were more frequently infected and able to transmit DEN-2 at higher rates than mainland populations. These data indicate that the Torres Strait region is potentially more receptive to dengue transmission than mainland localities, a finding discussed with respect to past outbreaks. PMID- 14765676 TI - Grasshoppers (Orthoptera: Acrididae) could serve as reservoirs and vectors of vesicular stomatitis virus. AB - Vesicular stomatitis (VS) is an economically devastating disease of livestock in the Americas. Despite strong circumstantial evidence for the role of arthropods in epizootics, no hematophagous vector explains the field evidence. Based on the spatiotemporal association of grasshopper outbreaks and VS epizootics, we investigated the potential role of these insects as vectors and reservoirs of the disease. The critical steps in the grasshopper-bovine transmission cycle were demonstrated, including 1) 62% of grasshoppers [Melanoplus sanguinipes (F.)] fed vesicular stomatitis virus (VSV) from cell culture became infected, with titers reaching 40,000 times the inoculative dose; 2) 40% of grasshoppers that cannibalized VSV-infected grasshopper cadavers became infected, amplifying virus up to 1,000-fold; 3) one of three cattle consuming VSV-infected grasshopper cadavers contracted typical VS and shed virus in saliva; and 4) 15% of grasshoppers became infected when fed saliva from this infected cow. The ecological conditions and biological processes necessary for these transmissions to occur are present throughout much of the Americas. Field studies will be required to show these findings are relevant to the natural epidemiology of VSV. PMID- 14765677 TI - Ixodes scapularis ticks (Acari: Ixodidae) from Louisiana are competent to transmit Borrelia burgdorferi, the agent of Lyme borreliosis. AB - The principal vector of Borrelia burgdorferi, the Lyme borreliosis spirochete, in the Northeast and Midwestern regions of the United States is the blacklegged tick Ixodes scapularis. Because of a favorable environment, I. scapularis is also plentiful in the South; however, a correlation with Lyme borreliosis cases does not exist in this region of the United States. Concern existed that something intrinsic to ticks found in Louisiana could mitigate their ability to transmit B. burgdorferi. Therefore, we set out to assess the ability of I. scapularis ticks from Louisiana to become infected with and transmit B. burgdorferi using mice as hosts. In the laboratory, mating adult female ticks collected in southeastern Louisiana were fed on the ears of rabbits. After oviposition and egg hatching, the resulting larvae were fed on mice that had been needle-inoculated with two different strains of B. burgdorferi sensu stricto, B31 and JD1. Larvae were found to be positive for spirochetes. Additional fed larvae were allowed to molt into the nymphal stage. Flat nymphs remained infected with B. burgdorferi. Infected nymphs were allowed to feed on naive mice, all of which became infected as shown by culture of ear biopsy specimens. Naive larvae were then fed on these same mice to assess transmissibility. The resulting engorged larvae harbored spirochetes. We have demonstrated that the I. scapularis ticks found in Louisiana are fully competent to carry and transmit B. burgdorferi infection. PMID- 14765678 TI - Experimental infection of California birds with western equine encephalomyelitis and St. Louis encephalitis viruses. AB - A total of 27 bird species from the San Joaquin and Coachella valleys of California were inoculated subcutaneously with sympatric strains of western equine encephalomyelitis (WEE) and St. Louis encephalitis (SLE) viruses. Overall, 133 of 164 birds inoculated with WEE virus developed a viremia detected by plaque assay; significantly greater than 72 of 163 birds inoculated with SLE virus. Host competence was calculated as the average number of days that each avian species had a viremia > or = 2 log10 plaque-forming units per 0.1 ml, the threshold for infecting susceptible Culex tarsalis Coquillett, the primary vector of these viruses in California. Eleven of 20 species inoculated with WEE virus had a value > or = 1 and were considered to be competent hosts, whereas only six of 22 species inoculated with SLE virus had a value > or = 1. Overall, 133 of 164 birds inoculated with WEE virus and 105 of 163 inoculated with SLE virus produced antibody detectable by enzyme immunoassay and/or plaque reduction neutralization test. Six birds infected with WEE virus (one house finch, three mourning doves, one Brewer's sparrow, and one white-crowned sparrow) and nine birds infected with SLE virus (two house finches, three white-crowned sparrows, one song sparrow, two Western scrub-jays, and one orange crowned warbler) contained viral RNA detected by reverse transcription-polymerase chain reaction at necropsy > 6 wk postinoculation; infectious WEE and SLE viruses were only recovered from three mourning doves and an orange-crowned warbler, respectively, after blind passage in mosquito cells. Our study indicated that birds with elevated field antibody prevalence rates may not be the most competent hosts for encephalitis viruses and that relatively few birds developed chronic infections that could be important in virus persistence and dispersal. PMID- 14765679 TI - Hairy maggot of Chrysomya villeneuvi (Diptera: Calliphoridae), a fly species of forensic importance. AB - We reported on the hairy maggot of Chrysomya villeneuvi Patton, collected from a human corpse in Thailand. Although the general morphology of the third instar closely resembled the more common hairy maggot blow fly, Chrysomya rufifacies (Macquart), the spines along the tubercles of each body segment could be used as a feature to distinguish between these species, with those of C. villeneuri bearing sharp-ended spines that encircle the entire tubercle. Not only has the resemblance of a hairy appearance between C. villeneuvi and C. rufifacies larvae been documented but also their similarity in predacious feeding behavior. The differentiation between these two species is necessary for post-mortem interval estimation in the corpse. PMID- 14765680 TI - Evidence for selection of insecticide resistance due to insensitive acetylcholinesterase by carbamate-treated nets in Anopheles gambiae s.s. (Diptera: Culicidae) from Cote d'Ivoire. AB - Pyrethroid-treated nets are an efficient tool for reducing malaria transmission and morbidity. The recent evolution of pyrethroid resistance in several Anopheles species represents a major threat for the future success of roll back malaria in Africa. The possible use of nonpyrethroid insecticides, such as carbamates, on nets is a promising alternative solution because these insecticides are effective against susceptible and pyrethroid-resistant populations of Anopheles and Culex mosquitoes. Unfortunately, carbamate resistance as a result of insensitive acetylcholinesterase has recently been detected in Anopheles gambiae s.s. populations from Cote d'Ivoire. Using biochemical assays on surviving Anopheles mosquitoes from an experimental hut trial, we showed evidence for selection for an insensitive acetylcholinesterase mechanism by carbamate impregnated bednets. However, no such selection has been found with nets treated with pyrethroid alone or pyrethroid/carbamate "two-in-one" -treated nets. Because pyrethroid impregnated nets were suspected to select for the Kdr mutation in An. gambiae, we propose that use of two-in-one nets could be a promising alternative strategy for the management of insecticide resistance in malaria vectors. PMID- 14765681 TI - Wild habitat and related fauna of Panstrongylus lutzi (Reduviidae, Triatominae). AB - Although Panstrongylus lutzi (Neiva and Pinto) is currently the fifth most frequent species of Triatominae captured in artificial structures in Brazil, its silvatic habitat remains unknown. A survey of its natural ecotopes was performed in an area of Bahia State. P. lutzi nymphal instars and adults were detected in burrows of Dasypodidae. Silvatic habitat of four other endemic triatomine species of the caatinga (Triatoma pseudomaculata Correa and Espinola, Triatoma brasiliensis Neiva, Rhodnius neglectus Lent, and Psammolestes tertius Lent and Jurberg) was also recorded. PMID- 14765682 TI - Two digestive trypsins occur in three species of neotropical anophelines. AB - Trypsin activity increases in the midgut of Anopheles aquasalis, Anopheles albitarsis, and Anopheles darlingi after a bloodmeal. The activity returns to basal levels at the time the blood is completely digested. Affinity chromatography, reversed-phase high performance liquid chromatography (HPLC), and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) were used to sequentially purify the mosquito trypsins found in the midguts at 24 h after feeding. Aminoterminal sequencing of the purified trypsins showed the occurrence of two distinct trypsins in the midgut of each of the mosquitoes studied. The sequences obtained are similar to those of the trypsins of other hematophagous insects. PMID- 14765683 TI - Acarine infracommunities associated with the Mexican free-tailed bat, Tadarida brasiliensis mexicana (Chiroptera: Molossidae) in arid regions of Mexico. AB - The Mexican free-tailed bat, Tadarida brasiliensis mexicana, is one of the most widely distributed bats, and its range includes the whole Mexican territory. Ectoparasites of this bat have been the subject of isolated reports, but no studies of its community ecology have been conducted. The acarine infracommunities associated with this bat were analyzed, comparing bat populations from three arid regions of Mexico: an abandoned factory in Nombre de Dios, Durango; a cave in Santiago, Nuevo Leon; and a church in Concepcion del Oro, Zacatecas. The acarine infracommunity in Nuevo Le6n's bats exhibited the highest levels of diversity as reflected by a higher richness, a lower dominance, and a moderate and relatively homogeneous abundance in this locality in relation to the other two. This pattern is influenced by stable cave conditions relative to artificial habitats. Notwithstanding, further studies are required to determine whether or not different habitat conditions are a primary factor in the process of structuring the acari infracommunities. PMID- 14765684 TI - Evaluation of transovarial transmission and transmissibility of Ehrlichia chaffeensis (Rickettsiales: Anaplasmataceae) in Amblyomma americanum (Acari: Ixodidae). AB - It has long been assumed that Ehrlichia chaffeensis (Anderson, Dawson & Wilson), is not transmitted transovarially in the lone star tick vector Amblyomma americanum (L.). To test this hypothesis, three beagle dogs, Canis familiaris (L.) (Carnivora: Canidae), were subcutaneously infected with E. chaffeensis (Arkansas strain). Uninfected nymphal lone star ticks were placed on the infected dogs and allowed to feed to repletion. These nymphal ticks were allowed to molt, and five of five adult female ticks sampled were confirmed to be infected by polymerase chain reaction (PCR). Forty infected adult ticks, the majority of which were female with several males included to stimulate feeding, were then placed on two uninfected dogs. Fourteen females were removed early and the rest were allowed to feed to repletion. After feeding to repletion, the six remaining females detached and two of these females were tested preoviposition, whereas the remaining four were allowed to lay eggs. All six of these fully engorged females tested negative by PCR. The egg clutches laid by four engorged adult females, and the larvae that hatched from these eggs along with larvae from a previously untested egg clutch, were shown to be uninfected by PCR. This seems to support the long-held assumption of lack of transovarial transmission. Also of interest, we found that these PCR-confirmed infected adult ticks reared in the laboratory did not transmit E. chaffeensis to uninfected dogs. PMID- 14765685 TI - Raising the bar for healing environments. PMID- 14765686 TI - Resilience: a historical review of the construct. AB - Evolution of the construct of resilience from physiological and psychological research extends from the 1800s to the present. This review supports 3 observations: (1) the importance of a dynamic, interactive perspective for understanding resilience; (2) the complexity of the construct requires a holistic perspective; and (3) the importance of exposure to diverse experiences and educational perspectives for professional health care students. PMID- 14765687 TI - Female Alzheimer's patient caregivers share their strength. AB - Spirituality is a valuable aspect in providing holistic care to Alzheimer's disease (AD) caregivers. This descriptive study explored the experience of AD caregiving in 14 older-women. A sample of African American and Caucasian participants presented spirituality and religion as essential aspects of their caregiving. Taking charge, adjusting/coping, making sense of the situation, and looking into the future were the themes that reflected spiritual/religious issues. The caregivers shared similar religious practices, but differed in the way they used spirituality and religious beliefs to construct the meaning of their experience. Implications for holistic care include incorporation of these differences in nursing practice. PMID- 14765688 TI - Health beliefs and women's use of hormone replacement therapy. AB - This research, part of an Internet-based study, examined whether beliefs about menopause and the Health Belief Model (HBM) variables explained women's use of hormone replacement therapy (HRT). Data were collected via a Web site and 208 women, ages 40 to 60, enrolled in the study over an eight-week period. Logistic regression analysis revealed perceived barriers and confidence as significant in predicting which women used HRT. The project was conducted prior to findings from the Women's Health Initiative (WHI). Despite a reduced risk of osteoporotic fractures in women receiving combined HRT, the WHI reports the risks of HRT outweigh the benefits. PMID- 14765689 TI - Nurses enhance performance through prayer. AB - Prayer can be empowering for performance enhancement. Researchers drew conclusions from a larger study that employed triangulation of research methods and a random sample of 1000 nurses to describe spiritual perspectives, interventions, and attitudes of nurses in the United States. This article describes the nurses' prayerful activities. PMID- 14765690 TI - Equine-facilitated psychotherapy benefits students and children. AB - Baccalaureate nursing students who participated in equine-facilitated psychotherapy (EFP) clinical observation found that they could benefit as much from the program as the child clients. By identifying beneficial educational outcomes of this nontraditional learning assignment, the authors hope readers will explore similar possibilities for nurses at various stages of their professional development. PMID- 14765691 TI - Spiritual interventions: how, when, and why nurses use them. AB - Researchers have performed limited studies regarding what nurses believe spirituality can do for their patients, the spiritual services they have offered, and under what circumstances. Because much of the extant research has only examined nurses involved in terminal care at different hospitals, it remains unclear upon which shared ideas and practices might nursing staff create a culture of spiritual care within a hospital. To address this situation, this study reports findings from a survey of bedside nurses at a university hospital. PMID- 14765692 TI - Battling the addiction dragon. PMID- 14765693 TI - Multimodal behavior in a four neuron ring circuit: mode switching. AB - We study a four-neuron ring circuit comprised of oscillating burst-type neurons unidirectionally coupled via inhibitory synapses. Simple circuits of this type have been used previously to study gait patterns. The ring circuit itself is a variant of the basic reciprocal inhibition network, and it exhibits the property of multistability (multiple stable modes of behavior). That is, different gait modes can be achieved via appropriate initialization of and parameterization of this self-excited oscillatory network. We demonstrate three common gait modes with this circuit: the walk, the bound, and a slightly rotated trot mode. Attention is focused mainly on the mechanisms of rapidly and effectively switching between these modes. Our simulations suggest that neuron membrane dynamics, as well as synaptic junctional properties, strongly influence phase sensitivity in the network; each synapse is a combination of both and can be characterized by a transient phase response curve (PRC). We use the same bursting neuron model to characterize all network neurons, and shape different transient PRCs by using different synaptic properties. The characteristics of these PRCs determine the gait modes sustained in any network configuration, as well as, the ability to switch between modes. The mechanisms explored in this simple circuit, may find application in the switching of more complicated gait pattern networks, as well as, in the design of neuromorphic gait pattern circuits. PMID- 14765694 TI - Robust algorithm for estimation of time-varying transfer functions. AB - We introduce a new method to estimate reliable time-varying (TV) transfer functions (TFs) and TV impulse response functions. The method is based on TV autoregressive moving average models in which the TV parameters are accurately obtained using the optimal parameter search method which we have previously developed. The new method is more accurate than the recursive least-squares (RLS), and remains robust even in the case of significant noise contamination. Furthermore, the new method is able to track dynamics that change abruptly, which is certainly a deficiency of the RLS. Application of the new method to renal blood pressure and flow revealed that hypertensive rats undergo more complex and TV autoregulation in maintaining stable blood flow than do normotensive rats. This observation has not been previously revealed using time-invariant TF analyses. The newly developed approach may promote the broader use of TV system identification in studies of physiological systems and makes linear and nonlinear TV modeling possible in certain cases previously thought intractable. PMID- 14765695 TI - Prediction of myelinated nerve fiber stimulation thresholds: limitations of linear models. AB - Computer models of neurons are used to simulate neural behavior, and are important tools for designing neural prostheses. Computation time remains an issue when simulating large numbers of neurons or applying models to real time applications. Warman et al. developed a method to predict excitation thresholds for axons using linear models and a predetermined critical voltage. We calculated threshold prediction error as a function of the location of an extracellular electrode using two different axon models to examine further threshold prediction using linear models. Threshold prediction error was low (<3% error) under the conditions examined by Warman et al., but under more general conditions, threshold prediction error was as high as 23.6%. Linear models were limited as effective tools for single fiber threshold prediction because accuracy was dependent on the nonlinear and linear models used, and any parameter that affected the extracellular potential distribution. Threshold prediction could be improved by appropriately choosing the membrane conductance of the linear model, but determination of an optimal conductance was computationally expensive. Finally, although single fiber threshold prediction error was partially masked when considering the input-output (I/O) properties of populations of axons, relatively large errors still occurred in population I/O curves generated with linear models. PMID- 14765696 TI - Identification of Hammerstein models with cubic spline nonlinearities. AB - This paper considers the use of cubic splines, instead of polynomials, to represent the static nonlinearities in block structured models. It introduces a system identification algorithm for the Hammerstein structure, a static nonlinearity followed by a linear filter, where cubic splines represent the static nonlinearity and the linear dynamics are modeled using a finite impulse response filter. The algorithm uses a separable least squares Levenberg-Marquardt optimization to identify Hammerstein cascades whose nonlinearities are modeled by either cubic splines or polynomials. These algorithms are compared in simulation, where the effects of variations in the input spectrum and distribution, and those of the measurement noise are examined. The two algorithms are used to fit Hammerstein models to stretch reflex electromyogram (EMG) data recorded from a spinal cord injured patient. The model with the cubic spline nonlinearity provides more accurate predictions of the reflex EMG than the polynomial based model, even in novel data. PMID- 14765697 TI - Automated feature extraction in color retinal images by a model based approach. AB - Color retinal photography is an important tool to detect the evidence of various eye diseases. Novel methods to extract the main features in color retinal images have been developed in this paper. Principal component analysis is employed to locate optic disk; A modified active shape model is proposed in the shape detection of optic disk; A fundus coordinate system is established to provide a better description of the features in the retinal images; An approach to detect exudates by the combined region growing and edge detection is proposed. The success rates of disk localization, disk boundary detection, and fovea localization are 99%, 94%, and 100%, respectively. The sensitivity and specificity of exudate detection are 100% and 71%, correspondingly. The success of the proposed algorithms can be attributed to the utilization of the model based methods. The detection and analysis could be applied to automatic mass screening and diagnosis of the retinal diseases. PMID- 14765698 TI - Nonlinear analysis of the separate contributions of autonomic nervous systems to heart rate variability using principal dynamic modes. AB - This paper introduces a modified principal dynamic modes (PDM) method, which is able to separate the dynamics of sympathetic and parasympathetic nervous activities. The PDM is based on the principle that among all possible choices of expansion bases, there are some that require the minimum number of basis functions to achieve a given mean-square approximation of the system output. Such a minimum set of basis functions is termed PDMs of the nonlinear system. We found that the first two dominant PDMs have similar frequency characteristics for parasympathetic and sympathetic activities, as reported in the literature. These results are consistent for all nine of our healthy human subjects using our modified PDM approach. Validation of the purported separation of parasympathetic and sympathetic activities was performed by the application of the autonomic nervous system blocking drugs atropine and propranolol. With separate applications of the respective drugs, we found a significant decrease in the amplitude of the waveforms that correspond to each nervous activity. Furthermore, we observed near complete elimination of these dynamics when both drugs were given to the subjects. Comparison of our method to the conventional low frequency/high-frequency ratio shows that our proposed approach provides more accurate assessment of the autonomic nervous balance. Our nonlinear PDM approach allows a clear separation of the two autonomic nervous activities, the lack of which has been the main reason why heart rate variability analysis has not had wide clinical acceptance. PMID- 14765699 TI - Sliding mode closed-loop control of FES: controlling the shank movement. AB - Functional electrical stimulation (FES) enables restoration of movement in individuals with spinal cord injury. FES-based devices use electric current pulses to stimulate and excite the intact peripheral nerves. They produce muscle contractions, generate joint torques, and thus, joint movements. Since the underlying neuromuscular-skeletal system is highly nonlinear and time-varying, feedback control is necessary for accurate control of the generated movement. However, classical feedback/closed-loop control algorithms have so far failed to provide satisfactory performance and were not able to guarantee stability of the closed-loop system. Because of this, only open-loop controlled FES devices are in clinical use in spite of their limitations. The purpose of the reported research was to design a novel closed-loop FES controller that achieves good tracking performance and guarantees closed-loop stability. Such a controller was designed based on a mathematical neuromuscular-skeletal model and is founded on a sliding mode control theory. The controller was used to control shank movement and was tested in computer simulations as well as in actual experiments on healthy and spinal cord injured subjects. It demonstrated good robustness, stability, and tracking performance properties. PMID- 14765701 TI - Predicting auditory tone-in-noise detection performance: the effects of neural variability. AB - Collecting and analyzing psychophysical data is a fundamental mechanism for the study of auditory processing. However, because this approach relies on human listening experiments, it can be costly in terms of time and money spent gathering the data. The development of a theoretical, model-based procedure capable of accurately predicting psychophysical behavior could alleviate these issues by enabling researchers to rapidly evaluate hypotheses prior to conducting experiments. This approach may also provide additional insight into auditory processing by establishing a link between psychophysical behavior and physiology. Signal detection theory has previously been combined with an auditory model to generate theoretical predictions of psychophysical behavior. Commonly, the ideal processor outperforms human subjects. In order for this model-based technique to enhance the study of auditory processing, discrepancies must be eliminated or explained. In this paper, we investigate the possibility that neural variability, which results from the randomness inherent in auditory nerve fiber responses, may explain some of the previously observed discrepancies. In addition, we study the impact of combining information across nerve fibers and investigate several models of multiple-fiber signal processing. Our findings suggest that neural variability can account for much, but not all, of the discrepancy between theoretical and experimental data. PMID- 14765700 TI - A new spatiotemporal regularization approach for reconstruction of cardiac transmembrane potential patterns. AB - The single-beat reconstruction of electrical cardiac sources from body-surface electrocardiogram data might become an important issue for clinical application. The feasibility and field of application of noninvasive imaging methods strongly depend on development of stable algorithms for solving the underlying ill-posed inverse problems. We propose a novel spatiotemporal regularization approach for the reconstruction of surface transmembrane potential (TMP) patterns. Regularization is achieved by imposing linearly formulated constraints on the solution in the spatial as well as in the temporal domain. In the spatial domain an operator similar to the surface Laplacian, weighted by a regularization parameter, is used. In the temporal domain monotonic nondecreasing behavior of the potential is presumed. This is formulated as side condition without the need of any regularization parameter. Compared to presuming template functions, the weaker temporal constraint widens the field of application because it enables the reconstruction of TMP patterns with ischemic and infarcted regions. Following the line of Tikhonov regularization, but considering all time points simultaneously, we obtain a linearly constrained sparse large-scale convex optimization problem solved by a fast interior point optimizer. We demonstrate the performance with simulations by comparing reconstructed TMP patterns with the underlying reference patterns. PMID- 14765702 TI - On the robustness of stride frequency estimation. AB - The robustness of stride frequency estimation (location and spread) from stride period data is investigated using influence functions. Theoretical analysis reveals that stride frequency estimates by Stokes et al. and by direct calculation have unbounded influence functions and zero breakdown points, implying a lack of both local and global robustness. Comparison of estimates obtained from an ensemble of pathological gait stride time series shows that on average, differences among estimators are not statistically significant (p > or = 0.59) for long time series (hundreds of strides). Specific circumstances under which nonrobust estimates depart from robust estimates are investigated in terms of outlier influence. We recommend some heuristic rules-of-thumb for prudent selection of nonrobust stride frequency estimators for a given stride time series. The theoretical and empirical estimator comparisons suggest that in general, more research on estimator robustness in quantitative gait analysis is warranted. PMID- 14765703 TI - Mining reproducible activation patterns in epileptic intracerebral EEG signals: application to interictal activity. AB - The study of interictal transient events may substantially complement the analysis of seizures in the presurgical evaluation of intractable epilepsy. A comprehensive methodology of quantifying reproducibility of activation patterns in intracerebral electroencephalography signals is presented. It may be applied to various forms of transient epileptic events under the assumption that a time of occurrence may be assigned to them. In this paper, the method is used on two different forms of interictal events (interictal spikes or sharpwaves and transient bursts of fast activity). The methodology is based on signal processing and data mining algorithms and proceeds in three steps: 1) detection of transient paroxysmal events (monochannel event); 2) identification of quasisynchronous transient paroxysmal events (multichannel events); and 3) automatic extraction of similar activation patterns. Results show that the methodology allows reproducible sequential activation sets to be identified from signals recorded in four patients. Potential advantages of the method are discussed with respect to other approaches. PMID- 14765704 TI - Vessel boundary tracking for intravital microscopy via multiscale gradient vector flow snakes. AB - Due to movement of the specimen, vasodilation, and intense clutter, the intravital location of a vessel boundary from video microscopy is a difficult but necessary task in analyzing the mechanics of inflammation and the structure of the microvasculature. This paper details an active contour model for vessel boundary detection and tracking. In developing the method, two innovations are introduced. First, the B-spline model is combined with the gradient vector flow (GVF) external force. Second, a multiscale gradient vector flow (MSGVF) is employed to elude clutter and to reliably localize the vessel boundaries. Using synthetic experiments and video microscopy obtained via transillumination of the mouse cremaster muscle, we demonstrate that the MSGVF approach is superior to the fixed-scale GVF approach in terms of boundary localization. In each experiment, the fixed scale approach yielded at least a 50% increase in root mean squared error over the multiscale approach. In addition to delineating the vessel boundary so that cells can be detected and tracked, we demonstrate the boundary location technique enables automatic blood flow velocity computation in vivo. PMID- 14765705 TI - Design and implementation of a hands-free electrolarynx device controlled by neck strap muscle electromyographic activity. AB - The electrolarynx (EL) voice prosthesis is widely used, but suffers from the inconvenience of requiring manual control. Therefore, a hands-free EL triggered by neck muscle electromyographic (EMG) activity was developed (EMG-EL). Signal processing circuitry in a belt-mounted control unit transforms EMG activity into control signals for initiation and termination of voicing. These control signals are then fed to an EL held against the neck by an inconspicuous brace. Performance of the EMG-EL was evaluated by comparison to normal voice, manual EL voice, and tracheo-esophageal (TE) voice in a series of reaction time experiments in seven normal subjects and one laryngectomy patient. The normal subjects produced voice initiation with the EMG-EL that was as fast as both normal voice and the manual EL. The laryngectomy subject produced voice initiation that was slower than with the manual EL, but faster than with TE voice. Voice termination with the EMG-EL was slower than normal voice for the normal subjects, but not significantly different than with the manual EL. The laryngectomy subject produced voice termination with the EMG-EL that was slower than with TE or manual EL. The EMG-EL threshold was set at 10% of the range of vocal-related EMG activity above baseline. Simulations of EMG-EL behavior showed that the 10% threshold was not significantly different from the optimum threshold produced through the process of error minimization. The EMG-EL voice reaction time appears to be adequate for use in a day-to-day conversation. PMID- 14765706 TI - Fluorescence imaging of electrical activity in cardiac cells using an all-solid state system. AB - Tracking spatial and temporal determinants of cardiac arrhythmogenesis at the cellular level presents challenges to the optical mapping techniques employed. In this paper, we describe a compact system combining two nontraditional low-cost solutions for excitation light sources and emission filters in fluorescence measurements of transmembrane potentials, Vm, or intracellular calcium, [Ca2+]i in cardiac cell networks. This is the first reported use of high-power blue and green light emitting diodes (LEDs), to excite cell monolayers stained with Vm - (di-8-ANEPPS) or [Ca2+]i - (Fluo-3) sensitive dyes. In addition, we use simple techniques for fabrication of suitable thin emission filters with uniform properties, no auto-fluorescence, high durability and good flexibility for imaging Vm or [Ca2+]i. The battery-operated LEDs and the fabricated emission filters, integrated with a fiber-optic system for contact fluorescence imaging, were used as tools to characterize conduction velocity restitution at the macro scale. The versatility of the LEDs for illumination is further emphasized through 1) demonstration of their usage for epi-illumination recordings at the single cell level, and 2) demonstration of their unique high-frequency light modulation ability. The LEDs showed excellent stability as excitation light sources for fluorescence measurements; acceptable signal-to-noise ratio and negligible cell photodamage and indicator dye photobleaching were observed. PMID- 14765707 TI - A multiconductance silicon neuron with biologically matched dynamics. AB - We have designed, fabricated, and tested an analog integrated-circuit architecture to implement the conductance-based dynamics that model the electrical activity of neurons. The dynamics of this architecture are in accordance with the Hodgkin-Huxley formalism, a widely exploited, biophysically plausible model of the dynamics of living neurons. Furthermore the architecture is modular and compact in size so that we can implement networks of silicon neurons, each of desired complexity, on a single integrated circuit. We present in this paper a six-conductance silicon-neuron implementation, and characterize it in relation to the Hodgkin-Huxley formalism. This silicon neuron incorporates both fast and slow ionic conductances, which are required to model complex oscillatory behaviors (spiking, bursting, subthreshold oscillations). PMID- 14765708 TI - Design of an integrated sensor for in vivo simultaneous electrocontractile cardiac mapping. AB - While there is extensive mapping of the spread of electrical activity in the heart, there have been no measurements of electrical and localized mechanical, or contractile, activity. Yet the development of effective treatments for diseases like chronic heart failure and cardiac hypertrophy depend on the ability to quantify improvements in electrocontractile function. In this paper, we present a sensor that is capable of making simultaneous, electrocontractile measurements. Its small size facilitates placement in multiple myocardial sites for multichannel studies. Semiconductor strain gages are used for force sensing, and Ag/AgCl-plated tungsten electrodes act as electrogram sensors. The sensor contains electronics on-board, including instrumentation amplifiers and a microprocessor for data sampling and analog-to-digital conversion. Each sensor can accurately detect 0-245+/-5 mV in two electrogram channels with a sensitivity of 0.96+/-0.2 mV/step and less than 2% error, and 0-144+/-29 g of contractile force with a sensitivity of 0.56+/-0.11 g/step in the analog-to-digital conversion and less than 6% error. The sensor has been tested in vivo in open chest rabbit and pig mapping studies. These studies indicated that the average peak-to-peak contractile force at the apex is smaller in the rabbit than the pig (13.3 versus 40.3 g), that the average peak-to-peak contractile force in the pig is smaller near the base than near the apex (31.3 versus 40.3 g), and that contractile force is visibly decreased during ventricular fibrillation compared to normal sinus rhythm. PMID- 14765709 TI - A new method for noninvasive measurement of multilayer tissue conductivity and structure using divided electrodes. AB - This paper outlines a new method for measuring multilayer tissue conductivity and structure by using divided electrodes, in which current electrodes are divided into several parts. Our purpose is to estimate the multilayer tissue structure and the conductivity distribution in a cross section of the local tissue by using bioresistance data measured noninvasively. The effect of the new method is assessed by computer simulations using a typical two-dimensional (2-D) model. In this paper, the conductivity distribution in the model is analyzed based on a finite difference method (FDM) and a steepest descent method (SDM). Simulation results show that the conductivity values of skin, fat, and muscle layers can be estimated with an error of less than 0.1%. When random noise at various levels is added to the measured resistance values, estimates of the conductivity values for skin, fat, and muscle layers are still reasonably precise: their root mean square errors are about 1.06%, 1.39%, and 1.61% for 10% noise. In a 2-D model, increasing the number of divided electrodes permits simultaneous estimates of tissue structure and conductivity distribution. Optimal configuration for divided electrodes is examined in terms of dividing pattern. PMID- 14765710 TI - Kinetics of thermal damage to a collagenous membrane under biaxial isotonic loading. AB - Prior isothermal uniaxial isotonic tests on tendons reveal that higher temperatures hasten the rate of thermal denaturation whereas larger mechanical loads delay it; moreover, these findings suggest a time-temperature-load equivalency whereby similar levels of denaturation, as reflected by tissue shrinkage, can be attained via many combinations of heating time, temperature level, and mechanical loading. Yet, most tissues and organs experience multiaxial loads in vivo, and their microstructure differs significantly from that of tendons, thus, we must also evaluate the effects of multiaxial stresses on the kinetics of denaturation in other tissues. In this paper, we describe a new experimental approach for performing isothermal biaxial isotonic tests on thin sheet-like specimens and we report effects of various thermomechanical loads on the rate and amount of multiaxial shrinkage of bovine epicardium. Consistent with uniaxial studies, epicardial shrinkage generally increased sigmoidally with heating time, and a characteristic heating time revealed increases in the rate of shrinkage with higher temperature and decreases with larger biaxial loads. Although this characteristic time exhibited an Arrhenius-type character, time temperature-load equivalency was not obtained when scaling time with this metric. General multiaxial thermomechanics is thus too complex to explain via a simple extension of uniaxial findings on tendons and there is a pressing need for more data and an appropriate theoretical framework. PMID- 14765712 TI - A robust DSP integrator for accelerometer signals. AB - The need for an accepted method for determining position data from digital accelerometer readings with known frequency range is very important. The method of this paper uses spectral information and provides more stability and accuracy than classic methods for the DSP case. It even reduces to a classic method for the nonoscillatory case. Examples show the robustness of the method, overcoming the instability apparent with other integration methods. PMID- 14765711 TI - Automatic detection of voice impairments by means of short-term cepstral parameters and neural network based detectors. AB - It is well known that vocal and voice diseases do not necessarily cause perceptible changes in the acoustic voice signal. Acoustic analysis is a useful tool to diagnose voice diseases being a complementary technique to other methods based on direct observation of the vocal folds by laryngoscopy. Through the present paper two neural-network based classification approaches applied to the automatic detection of voice disorders will be studied. Structures studied are multilayer perceptron and learning vector quantization fed using short-term vectors calculated accordingly to the well-known Mel Frequency Coefficient cepstral parameterization. The paper shows that these architectures allow the detection of voice disorders--including glottic cancer--under highly reliable conditions. Within this context, the Learning Vector quantization methodology demonstrated to be more reliable than the multilayer perceptron architecture yielding 96% frame accuracy under similar working conditions. PMID- 14765713 TI - Internodal myelinated segments: delay and RGC time-domain Green function model. AB - The myelinated axon can be modeled by means of a distributed RGC circuit. The Green function of this model allows for a generic formulation of the internodal segment response to any kind of stimulus, and accounts for the delay of the action potential associated with this segment. The RGC model accuracy is comparable to that of a more complex electromagnetic model, and predicted delay agrees with experimental measurements. PMID- 14765714 TI - Creating a vision for our preferred future. PMID- 14765715 TI - Ethical concerns for organ transplant coordinators. AB - A great deal has been written about the ethics of organ transplantation. Much of it has (appropriately) concerned issues of policy--for example, how best to allocate scarce organs, avoid conflicts of interest, and increase the pool of potential donors. Although these discussions are important, they often fail to provide much context for applying relevant ethical precepts to the day-to-day activities of those directly involved in organ procurement. The purpose of this paper is to provide such a context by reviewing several key conceptual issues, drawing attention to various ethical and procedural distinctions, and offering some practical recommendations for addressing these issues with physicians and other healthcare providers. PMID- 14765716 TI - Advanced assessment of hemodynamic parameters during donor care. AB - Detailed evaluation of cardiovascular function by hemodynamic "profiling" during donor care may be necessary to select appropriate treatment for hypotension. Preload, systemic vascular resistance, heart rate, and contractility are all determinants of cardiac output and blood pressure. This review discusses the measurement, physiological limitations, and pharmacological manipulation of those variables during care of selected donors. PMID- 14765717 TI - The tissue donation experience: a comparison of donor and nondonor families. AB - CONTEXT: Although there has been an increase in studies examining the experiences of organ donor and nondonor families, scant empirical research specifically concerning tissue donation has been conducted. OBJECTIVE: To assess the perceptions and attitudes of both donor and nondonor next of kin to whom a tissue donation request had been directed. DESIGN: Cross-sectional, 2-group analysis of a telephone survey. SETTING AND PARTICIPANTS: Participants were 507 adults who consented to tissue donation and 507 adults who refused consent to tissue donation following the death of a family member. All donation requests were made by telephone by trained staff. MAIN OUTCOME MEASURE: The primary outcome measures were demographic and contextual information surrounding the donation request, perceptions and attitudes toward tissue donation, and reasons for donation or refusal to donate. RESULTS: Findings suggest that most tissue donor family members were satisfied with the request and donation experience. Also, there were important differences in attitudes and perceptions between donor and nondonor family members, with nondonors reporting more negative attitudes and beliefs about tissue donation. PMID- 14765718 TI - Development of the University of Wisconsin donation After Cardiac Death Evaluation Tool. AB - BACKGROUND: Donation after cardiac death is a method by which severely neurologically injured patients not fulfilling brain-death criteria can donate organs. OBJECTIVE: To develop an evaluation tool that can be used to predict if a patient is a suitable candidate for donation after cardiac death. METHODS: The University of Wisconsin Donation After Cardiac Death Evaluation Tool assigns numeric values to observable clinical parameters to yield an overall predictive score of suitability for donation after cardiac death. This evaluation tool is typically utilized in a critical care unit to evaluate patients with a severe neurological injury, who do not meet brain-death criteria, and for whom the physician and family have chosen to terminally withdraw life support. Each patient is disconnected from a ventilator and observed for up to 10 minutes. Observations are then scored to yield a prediction of suitability for donation after cardiac death. RESULTS: Using the University of Wisconsin Donation After Cardiac Death Evaluation Tool, we were able to predict suitability for donation after cardiac death 83.7% of the time, within a 60-minute period and 74.4% of the time within a 120-minute period. The actual results using the tool were higher when clinical observations were included in the donation after cardiac death evaluation--an overall accuracy of 88.4%. PMID- 14765719 TI - Donation after cardiac death: two case studies. AB - Few transplant centers consider using lungs from cardiac death donors because of warm ischemic damage. In certain scenarios, the recovery and transplantation of lungs from cardiac death donors are appropriate. A young person with a severe neurologic and spinal cord injury, who is not brain dead and who is otherwise healthy, should be considered as a cardiac death donor. A protocol should be established with local lung transplant surgeons to facilitate the successful procurement of lungs from cardiac death donors. In addition, when patients present to hospital emergency rooms with nonsurvivable injuries either in cardiac arrest or with extremely labile vital signs, uncontrolled donation after cardiac death can be considered. It is important to obtain informed consent from the family and to suspend any previous do-not-resuscitate orders before initiating resuscitative efforts. If an organ procurement coordinator and team are within close proximity to the hospital, consideration should be given to uncontrolled donation after cardiac death. PMID- 14765720 TI - Impact of long-term immunosuppressive therapy on psychosocial and physical well being in liver transplant recipients. AB - To achieve the fullest potential of transplantation, continuing concern for the recipients' quality of life must be a part of the process. Database records of patients who are currently alive and received transplants between 1982 and 1991 were retrospectively analyzed. Recipients were contacted and asked to answer a quality-of-life questionnaire. Of 105 liver transplant recipients, 51 died within 10 years after transplantation; 47 were contacted. Posttransplant complications included hypertension (64%), posttransplant diabetes mellitus (17%), osteopenia (40%), osteoporosis (26%), and heart disease (17%). Most recipients reported all aspects of their life to be average, if not better than their age-matched peers. Although most recipients complained about side effects of immunosuppressive agents, they were all happy to be alive and agreed that their quality of life showed an impressive favorable change to a level exceeding that of the general population. These results suggest that liver transplantation not only improved survival but also quality of life. PMID- 14765721 TI - Minimal intervention programs for weight loss in heart transplant candidates: a preliminary examination. AB - CONTEXT: Overweight can be a contraindication for cardiac transplantation, and empirical findings suggest that obesity may pose serious posttransplant health risks that can increase morbidity and mortality. OBJECTIVE: This study assessed the relative effectiveness of 2 minimal intervention programs to assist weight loss in heart transplant candidates. DESIGN: A randomized trial was employed to assess changes in body weight. SETTING: A large, tertiary care hospital in a Southern locale. PATIENTS: Forty-three heart transplant candidates (74% men, 79% married, 86% white), with a mean pretreatment body mass index of 32.4 (SD = 4.4). INTERVENTIONS: Patients were randomly assigned to a bibliotherapy weight-loss program or a bibliotherapy plus telephone contact weight-loss program. MAIN OUTCOME MEASURES: Change in body weight over 3 months and return rate of measures of program adherence (3-day food diaries and self-monitoring postcards). RESULTS: An intent-to-treat analysis showed a significant weight loss at posttreatment for the sample as a whole. Within-group analyses indicated that a significant weight change (P < .05) in the telephone contact group (mean [SD] = -2.76 [4.96] kg) but not the bibliotherapy-only group (mean [SD] = -1.02 [2.97] kg). Participants in the telephone contact group returned more 3-day food diaries and self-monitoring postcards, with pounds lost significantly correlated with the number of completed self-monitoring postcards. These findings suggest that a minimal intervention program involving information plus limited professional contact may represent a viable approach to assisting overweight transplant candidates in weight management. PMID- 14765722 TI - Development of a questionnaire to assess attitudes and beliefs toward organ donation among Asians. AB - In the United Kingdom, people from the Indian subcontinent have a higher incidence of kidney failure, are less likely to receive a kidney transplant, and have a higher mortality rate from renal disease. In addition, Asians are reluctant donors; white people make up 95% of the donor pool. We developed a questionnaire to determine the knowledge and attitudes toward organ donation in the Asian communities. The setting was a public forum; the aims were (1) to study the attitudes of Asians toward organ donation, (2) to validate our questionnaire, and (3) to initiate a campaign of increasing awareness of organ donation and transplantation in the Asian community in the greater Glasgow area. Our survey suggested that Asians have an awareness of organ donation and transplantation, and they would donate an organ to a family member; however, they did not carry a donor card. Respondents unanimously agreed that lobbying by religious leaders was the best means to promote awareness. PMID- 14765723 TI - Transplant coordination program: a useful tool to improve organ donation in Venezuela. AB - Donor shortage is the single most important limitation for allowing adequate growth of transplant programs. Transplant coordination programs have been shown to provide solutions to this situation worldwide. To evaluate the efficacy of transplant coordination programs in Venezuela, a pilot program was implemented at a 1200-bed teaching hospital. The implementation of this program included an assessment of the hospital's donation practices such as donor identification, maintenance, brain-death diagnosis, family consent for donation, and timely transport and allocation of organs and tissues. A follow-up 1 year after the implementation of the transplant coordination program demonstrated a 7-fold increase in the number of donors compared with the 2 previous years when the program did not exist. During the first year of application, the transplant coordination program resulted in solutions in how to address issues surrounding the procurement process in a hospital with a high potential donor rate; a linkage between the coordinator and the medical staff through educational activities; increased skills of hospital staff; and a methodology that should be applied extensively in hospitals with high donor potential to deal with the organ shortage. PMID- 14765724 TI - Coping with transplantation. Part 1: The pretransplant period. PMID- 14765725 TI - Special editorial--internal medicine and clinical pathology: rebuilding the relationship. PMID- 14765726 TI - Antimicrobial use in the treatment of calf diarrhea. AB - Calves with diarrhea often have small intestinal overgrowth with Escherichia coli bacteria, regardless of the inciting cause for the diarrhea, and 30% of systemically ill calves with diarrhea have bacteremia, predominantly because of E coli. Antimicrobial treatment of diarrheic calves should therefore be focused against E coli in the small intestine and blood, the 2 sites of infection. Fecal bacterial culture and antimicrobial susceptibility testing is not recommended in calves with diarrhea because fecal bacterial populations do not accurately reflect small intestinal or blood bacterial populations and because the break points for susceptibility test results have not been validated. Antimicrobial efficacy is therefore best evaluated by the clinical response of a number of calves to treatment, with calves randomly assigned to treatment groups. Amoxicillin, chlortetracycline, neomycin, oxytetracycline, streptomycin, sulfachloropyridazine, sulfamethazine, and tetracycline administered PO are currently labeled in the United States for the treatment of calf diarrhea. On the basis of published evidence for the oral administration of these antimicrobial agents, only amoxicillin can be recommended for the treatment of diarrhea. Dosage recommendations are amoxicillin trihydrate (10 mg/kg PO q12h) or amoxicillin trihydrate-clavulanate potassium (12.5 mg combined drug/kg PO q12h) for at least 3 days; the latter constitutes extra-label drug use. Parenteral administration of broad-spectrum beta-lactam antimicrobials--ceftiofur (2.2 mg/kg IM or SC q12h) and amoxicillin or ampicillin (10 mg/kg IM q12h)--or potentiated sulfonamides (25 mg/kg IV or IM q24h) is recommended for treating calves with diarrhea and systemic illness; both constitute extra-label drug use. In calves with diarrhea and no systemic illness (normal appetite for milk, no fever), it is recommended that the health of the calf be monitored and that oral or parenteral antimicrobials not be administered. PMID- 14765727 TI - Evaluation of the oral antihyperglycemic drug metformin in normal and diabetic cats. AB - Metformin is an oral antidiabetic drug that improves control of glycemia primarily by inhibiting hepatic gluconeogenesis and glycogenolysis. This study evaluated the usefulness of metformin for the treatment of diabetes mellitus in cats. The study consisted of 3 phases. Phase I was a dose-finding study performed in healthy cats that were randomly administered varying doses of metformin to determine the approximate dose that would yield plasma concentrations known to be effective in humans. Phase 2 was a 3-week safety study performed in healthy cats to determine if cats could tolerate the daily oral dose and administration protocol identified during phase 1. Phase 3 was a clinical trial evaluating the clinical response of diabetic cats to oral metformin treatment. Five cats with newly diagnosed, naturally acquired diabetes mellitus were enrolled in phase 3. Plasma metformin concentrations in the therapeutic range of 0.5-2 microg/mL were achieved with doses of 50 mg/cat PO q12h without dramatic drug accumulation. Intermittent lethargy, inappetence, vomiting, and weight loss were identified, and the results of the CBC, serum biochemical analysis, plasma lactate concentration, and urinalysis remained within the reference range during phase 2 of the study. During phase 3, control of glycemia was achieved in 1 of 5 diabetic cats after 8 weeks of metformin treatment; 3 cats failed to respond to metformin, and treatment with insulin was initiated after 7-8 weeks of metformin treatment; 1 cat died unexpectedly 11 days after starting metformin treatment. The cause of death was not determined. The serum insulin concentration was within or greater than the reference range in the responder diabetic cat and was undetectable or at the low end of the reference range in the nonresponder diabetic cats. The results of this study suggest that metformin is beneficial only in those diabetic cats with detectable concentrations of insulin at the time metformin treatment is initiated. PMID- 14765728 TI - Lymphocyte blastogenic responses to inciting food allergens in dogs with food hypersensitivity. AB - Lymphocyte blastogenic responses against food allergens in dogs with food hypersensitivity were evaluated in this study. Eleven dogs with food hypersensitivity, based on food elimination and oral food provocation tests and allergic responses to food allergens, were examined by various tests such as intradermal testing, antigen-specific IgE testing, and lymphocyte blastogenic responses. The number and kinds of food allergens identified as positive by these tests were compared with the offending food allergens that were found in an oral food provocation test. In 9 (82%) of the 11 dogs with food hypersensitivity, there was close agreement for positive allergens between the results of lymphocyte blastogenic responses and oral food provocation test; however, there was little agreement for intradermal and IgE testing of the positive allergens with those of the oral food provocation test (11% and 31%, respectively). In the 9 dogs, the stimulation indices of lymphocyte blastogenic responses increased to 2.0-10.1 upon food provocation but decreased significantly to 0.7-1.4 upon feeding the elimination diet until clinical signs disappeared. These results indicate that lymphocyte blastogenic responses may fluctuate because of exposure to offending food allergens in dogs with food hypersensitivity. Lymphocytes reactive to food allergens may play an important role in the pathogenesis of food hypersensitivity in dogs. PMID- 14765729 TI - Management of advanced tracheal collapse in dogs using intraluminal self expanding biliary wallstents. AB - Twenty-four client-owned dogs with tracheal collapse refractory to conventional treatment underwent management with an intraluminal self-expanding stainless steel endoprosthesis (Wallstent). Initial improvement of clinical signs was observed in 95.8% of the dogs. Two dogs (8.3%) died within a median interval of 6 days after stent implantation due to incorrect placement and size of the stent and emphysema, respectively. A dry cough occurred temporarily in most of the patients. One dog each (4.1%) suffered mild transient tracheal hemorrhage and pneumomediastinum. The results showed that the initial survival rate of intraluminal stabilization was comparable with surgical implantation of extratracheal prostheses. Clinical reevaluation was performed in 18 dogs within a median interval of 68 days after treatment. Of the dogs treated, 30.4% were reported to be asymptomatic after stent implantation, 60.9% improved markedly, and 4.3% remained symptomatic. In all patients undergoing endoscopy, the Wallstents were almost completely covered with tracheal epithelium. A median shortening of 27.3% of the endoprosthesis within a median interval of 175 days after stent implantation in 15 of 18 dogs was noted. The shortening was associated with clinical signs in 2 patients. In 5 dogs, steroid-responsive granuloma formation resulted in a severe reduction of the tracheal lumen in 3 patients. The results suggest that implantation of Wallstents was minimally invasive and provided stabilization of collapsed thoracic tracheal portions in addition to the cervical part of the trachea. This minimally invasive method for the management of severe tracheal collapse therefore provides an attractive alternative to surgery. PMID- 14765730 TI - Systemic absorption of amitriptyline and buspirone after oral and transdermal administration to healthy cats. AB - A prospective study was performed to determine the relative availability of buspirone and amitriptyline after oral and transdermal routes of administration in 6 adult cats. For topical administration, drugs were compounded in a transdermal organogel containing pluronic and lecithin (PLO). Using a crossover design, each cat received a single dose of amitriptyline (5 mg) and buspirone (2.5 mg) by the transdermal and oral route of administration with at least a 2 week washout interval between drug treatments. Blood samples were obtained at 0, 0.5, 1, 2, 4, 6, 8, 10, and 12 hours after drug administration for determination of plasma drug concentrations. Plasma concentrations of immunoreactive amitriptyline and buspirone were determined using commercial enzyme-linked immunosorbent assay (ELISA) tests. Systemic absorption of amitriptyline and buspirone administered by the transdermal route was poor compared with the oral route of administration. Until supporting pharmacokinetic data are available, veterinarians and cat owners should not rely on the transdermal route of administration for treating cats with amitriptyline or buspirone. PMID- 14765731 TI - Antinuclear antibodies can be detected in dog sera reactive to Bartonella vinsonii subsp. berkhoffii, Ehrlichia canis, or Leishmania infantum antigens. AB - The presence of antinuclear antibodies (ANAs) is used to support a clinical diagnosis of systemic lupus erythematosus (SLE) in dogs. However, clinicians must interpret the detection of ANAs with caution, particularly in light of increasing evidence that dogs with known bacterial and protozoal infections can have high ANA titers. Retrospectively, medical records were reviewed for all dogs that were concurrently tested for antinuclear antigens and Bartonella vinsonii (berkhoffii), Ehrlichia canis, or Rickettsia rickettsii antigens between 1990 and 2000. When analyzed on the basis of reactivity to a specific infectious agent, 75% of the B vinsonii (berkhoffii) seroreactors, 16.7% of the E canis seroreactors, and 0% of the R rickettsii seroreactors had concurrent ANAs. Subsequent prospective testing did not detect ANAs in convalescent sera from dogs experimentally infected with B vinsonii (berkhoffii), E canis, or R rickettsii. However, 10-20% B vinsonii (berkhoffii), E canis, or Leishmania infantum reactive sera from naturally infected dogs contained ANAs. In addition, 45% of sera from dogs that are reactive to multiple vectorborne organisms were more likely to contain ANAs when compared to sera from dogs reactive to only 1 test antigen. When interpreting the relevance of seroreactivity to nuclear antigens, clinicians should recognize that dogs with seroreactivity to B vinsonii (berkhoffii), E canis, or L infantum antigens (especially those with seroreactivity to more than one of these pathogens) may produce ANAs. PMID- 14765732 TI - The effects of exercise on urinary albumin excretion in dogs. AB - Persistent microalbuminuria has been shown to be an indicator of glomerular damage associated with early progressive renal disease in people and dogs. In people, transient or reversible microalbuminuria has been shown to occur with exercise. A semi-quantitative test to measure microalbuminuria in the dog recently has become available. The purpose of this study was to determine if mild to-moderate exercise induced microalbuminuria in the dog. Twenty-six dogs were included in the study after undergoing tests to rule out hyperglycemia, urinary tract infection, azotemia, and a urine protein:creatinine ratio >1. Exercise consisted of 20 minutes of flat treadmill running. Urine samples were collected on 2 separate days before exercise, the morning of exercise, 3 hours postexercise, 7-9 hours postexercise, and each of the 2 mornings after exercise. For 24 of 26 dogs, this procedure was repeated after a minimum 7-day interval between exercise sessions. The canine E.R.D. (early renal disease)-Screen Urine Test (E.R.D.-Screen test) was used to determine semiquantitative urine albumin concentrations. Microalbuminuria-positive samples, as determined by the E.R.D. Screen test, were further analyzed to determine quantitative albumin concentrations. Four (15%) dogs were microalbuminuria positive. In each of these dogs, microalbuminuria was present both before and after exercise with no quantitative increase in urine albumin concentration postexercise. Twenty-two (85%) dogs were microalbuminuria negative throughout the study and did not develop microalbuminuria at any time after exercise. On a 95% confidence interval, the proportion of dogs that might be expected to develop microalbuminuria after exercise is between 0 and 15%. PMID- 14765733 TI - A prospective study of canine infective endocarditis in northern California (1999 2001): emergence of Bartonella as a prevalent etiologic agent. AB - A prospective study was performed (June 1999 to May 2001) to determine the incidence of infective endocarditis (IE) due to Bartonella in dogs in northern California and to compare these patients with other dogs with IE. IE was diagnosed antemortem based on clinical signs and echocardiography in 18 dogs. The etiologic agent was Bartonella sp. in 5 dogs (28%) and was diagnosed by high seroreactivity to Bartonella (titer > 1:512; range, 1:1,024-1:4,096); and confirmed postmortem by positive polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) from the infected valve and partial DNA sequencing of the citrate synthase gene (glt A). Conventional bacteria were causative agents in 7 dogs (39%). An etiologic agent was not identified in 6 dogs (33%). Bartonella vinsonii berkhoffii (n = 3), B clarridgeiae (n = 1), and a B clarridgeiae-like organism (n = 1) were identified. Blood culture was positive only for the IE case due to B clarridgeiae. All dogs with IE due to Bartonella were also seroreactive to Anaplasma phagocytophilum. All dogs with IE due to Bartonella had lesions only on the aortic valve. Of the cases of IE not due to Bartonella, 31% involved the aortic valve, 61% the mitral valve, and 8% both valves. Dogs with mitral valve IE lived longer than all dogs with aortic valve IE (P = .004) and dogs with IE of the aortic valve due to Bartonella (P = .002). In conclusion, Bartonella is a common cause of IE in dogs of northern California. A high Bartonella serologic titer (> 1:512) is useful antemortem to diagnose aortic valve IE due to Bartonella. PMID- 14765734 TI - Superficial necrolytic dermatitis in 11 dogs with a history of phenobarbital administration (1995-2002). AB - The clinical records of 11 dogs with histologically confirmed superficial necrolytic dermatitis (SND) and a history of phenobarbital (PB) administration (SND/PB) were evaluated retrospectively (1995-2002). Historical, clinical, clinicopathologic, ultrasonographic, and pathologic findings were compared with those in dogs with SND without prior PB exposure (SND/No PB; n = 9) and with those dogs with PB-associated hepatotoxicity without skin disease (PB/hepatotoxicity). Dogs in the SND/PB group accounted for 44% of all histologically confirmed cases of SND that were evaluated at The Ohio State University Veterinary Teaching Hospital between 1995 and 2002. Median age of dogs in the SND/PB group was 10 years, and median duration of PB therapy was 6 years. Mean alanine aminotransferase (ALT) activity was 239 U/L, and median duration of abnormally high ALT activity was 6.25 months before SND diagnosis. Plasma amino acid concentrations measured in 1 dog were severely decreased. Ultrasonographic findings of hypoechoic nodules with hyperechoic borders corresponded to pathologic findings of nodular areas of normal hepatic tissue surrounded by zones of collapsed parenchyma with vacuolated hepatocytes. Clinical, clinicopathologic, ultrasonographic, and pathologic features of SND/PB and SND/No PB were similar. PB-associated cirrhosis and overt hepatic failure were not features of SND/PB. Different pathogenic mechanisms might induce SND in dogs. Chronic administration of PB requires further examination as a potential risk factor for the development of SND. PMID- 14765735 TI - Hepatotoxicity associated with CCNU (lomustine) chemotherapy in dogs. AB - One hundred seventy-nine tumor-bearing dogs were treated with 1-(2-chloroethyl)-3 cyclohexyl-1-nitrosourea (CCNU) between 1995 and 2001. CCNU was given as a single dose of 50-110 mg/m2 body surface area PO. Treatment interval varied, but the minimal interval between CCNU doses was 3 weeks. After treatment, 11 dogs (6.1%) developed hepatic toxicity. The median number of CCNU doses and the median total cumulative CCNU dose were significantly higher in dogs that developed hepatic toxicity (4 doses; 350 mg/m2) than in dogs without hepatic damage (3 doses; 230 mg/m2). Median duration to detection of hepatic toxicity from the last dose of CCNU was 11 weeks (range 2-49 weeks). Common biochemical abnormalities were abnormally high serum liver enzyme activities and hypoalbuminemia. Six dogs with CCNU-associated hepatic toxicity had ascites, and 3 dogs had concurrent pleural effusion. Serum concentrations of bile acids were abnormally high in 4 of 5 dogs tested. Percutaneous ultrasound-guided liver biopsies were performed in 10 dogs, and findings were nonspecific and chronic in nature. Seven dogs were euthanized because of progressive liver failure, and their median survival from diagnosis of liver disease was 9 weeks. Three dogs died of other causes and 1 dog of unknown cause. Although clinical signs resolved in 3 dogs, biochemical abnormalities and histopathologic lesions persisted 4 to 38 months from the time of diagnosis of liver disease. Our findings suggest that CCNU can cause delayed, cumulative dose related, chronic hepatotoxicity that is irreversible and can be fatal. PMID- 14765736 TI - Clinical efficacy and safety of recombinant canine erythropoietin in dogs with anemia of chronic renal failure and dogs with recombinant human erythropoietin induced red cell aplasia. AB - The efficacy and safety of recombinant canine erythropoietin (rcEPO) therapy was evaluated in 19 dogs with anemia of chronic renal failure (group 1) and 6 dogs with chronic renal failure and recombinant human erythropoietin (rhEPO)-induced red cell aplasia (group 2). Hematocrit (Hct) and absolute reticulocyte count (ARC) were monitored weekly for the first 8 weeks, CBC (including ARC) and serum iron profiles were evaluated monthly, and serum biochemical analyses were performed every 2 months for 6 (group 2) to 12 (group 1) months. For group 1 dogs, median Hct and ARC increased significantly during the 1st week of rcEPO treatment, and median Hct was sustained at >35% after week 5. In contrast, median Hct and ARC for group 2 did not change significantly with rcEPO treatment, even with doses greater than those used in group 1. Nevertheless, 2 (33%) of the 6 dogs in group 2 developed erythroid hyperplasia, reticulocytosis, and increases in Hct with rcEPO treatment. Although median systolic blood pressure did not change significantly in either group, 5 dogs developed systolic blood pressures > or = 180 mm Hg during the study. Appetite and energy level improved in most group 1 dogs with increases in Hct. Recombinant cEPO stimulated erythrocyte production in dogs with nonregenerative anemia secondary to chronic renal failure without causing the profound erythroid hypoplasia that can occur in rhEPO-treated dogs. Unfortunately, rcEPO was not as effective in restoring erythrocyte production in dogs that had previously developed rhEPO-induced red cell aplasia. PMID- 14765737 TI - Airway mucus in recurrent airway obstruction--short-term response to environmental challenge. AB - Mucus accumulation and neutrophilic inflammation in the airways are hallmarks of heaves. Endoscopically visible mucus accumulations, however, have not been studied during exposure to dusty hay and allergens (ie, environmental challenge). We hypothesized that (1) heaves-affected horses have increased mucus accumulation compared with controls, (2) mucus accumulations increase in heaves-affected horses during environmental challenge, and (3) environmental challenge also induces neutrophilic inflammation and mucus accumulation in control horses. Mucus accumulation was graded endoscopically (mucus grades [MGs] 1-5), and airway inflammation was evaluated by bronchoalveolar lavage fluid (BALF) cytology before (0 hours) and during (6, 24, 48 hours) environmental challenge. Large amounts of mucus (MG 4-5) were specific for heaves-affected horses in this study. Variation among controls was considerable, however, and intermediate grades (MG 2-3) were nonspecific, showing complete overlap between the 2 groups. Median mucus accumulations (25th, 75th percentiles) increased in heaves-affected horses from MG 2.5 (1.5, 3.5) at baseline to MG 3.5 (2.0, 4.0), 4.0 (3.0, 4.0), and 4.0 (4.0, 4.0) at 6, 24, and 48 hours, respectively. MG values did not increase in controls -overall MG 1.0 (1.0, 2.0)--even though controls also showed a moderate increase of BALF neutrophils. Mucus accumulations before and especially after exposure to dust and allergens are increased in heaves-affected horses compared with controls. Healthy controls show considerable variability in mucus accumulation but, despite an influx of neutrophils into the airways, no increase of mucus accumulation after exposure to hay dust. PMID- 14765738 TI - Sarcocystis neurona-specific immunoglobulin G in the serum and cerebrospinal fluid of horses administered S neurona vaccine. AB - A vaccine against Sarcocystis neurona, which induces equine protozoal myeloencephalitis (EPM), has received conditional licensure in the United States. A major concern is whether the immunoglobulin G (IgG) response elicited by the vaccine will compromise the use of Western blotting (WB) as a diagnostic tool in vaccinated horses with neurologic disease. Our goals were to determine if vaccination (1) causes seroconversion: (2) causes at least a transient increase in S neurona-specific IgG in the cerebrospinal fluid (CSF); and (3) induces an IgG response that can be differentiated from that induced by natural exposure. Horses included in the study (n = 29) were older than 6 months with no evidence of neurologic disease. The presence or absence of anti-S neurona antibodies in the serum of each horse was determined by WB analysis. Seropositive horses had CSF collected and submitted for cytology, CSF index, and WB analysis. The vaccine was administered to all the horses and boostered 3-4 weeks later. On day 14 after the 2nd administration, serum and CSF were collected and analyzed. Eighty-nine percent (8 of 9) of the initial seronegative horses seroconverted after vaccination, of which 57% (4 of 7) had anti-S neurona IgG in their CSE Eighty percent (16 of 20) of the seropositive horses had an increase in serum S neurona IgG after vaccination. Of the 6 of 20 horses that were initially seropositive/CSF negative, 2 were borderline positive for anti-S neurona IgG in the CSF, 2 tested positive, and 2 were excluded because the CSF sample had been contaminated by blood. There were no WB banding patterns that distinguished samples from horses that seroconverted due to vaccination versus natural exposure. Caution must be used in interpreting WB analysis from neurologic horses that have been recently vaccinated for EPM. PMID- 14765739 TI - Effects of supplemental lactoferrin on serum lactoferrin and IgG concentrations and neutrophil oxidative metabolism in Holstein calves. AB - Lactoferrin (LF) is an iron-binding protein present in both colostrum and secondary granules of polymorphonuclear neutrophils (PMNs). We hypothesized that supplemental LF enhances neutrophil function in neonatal calves. Newborn calves were assigned to receive colostrum (C), colostrum + LF (CLF, 1 g/kg), or milk replacer + LF (MRLF, 1 g/kg). Serum (LF and IgG) and whole blood (neutrophil isolation) samples were obtained prior to treatment (day 0) and at 24 hours and 9 days of age. Serum IgG concentrations (mean +/- SD) in C, CLF, and MRLF calves at 24 hours were 1,911 +/- 994 mg/dL, 2,181 +/- 625 mg/dL, and 0 mg/ dL, respectively. Serum LF concentrations in C, CLF, and MRLF calves on day 0 were 324 +/- 334 ng/mL (range 0-863 ng/mL), 135 +/- 158 ng/mL (range 0-429 ng/mL), and 318 +/- 337 ng/mL (range 0-964 ng/mL), respectively. LF concentrations in C, CLF, and MRLF calves at 24 hours were significantly higher (P < .05), at 1,564 +/- 1,114 ng/mL (range 335-3,628 ng/mL, 2,237 +/- 936 ng/mL (range 31-3,287 ng/mL), and 3,189 +/- 926 ng/mL (range 1,736-4,120 ng/mL), respectively. Cytochrome c reduction in opsonized zymosan-treated or phorbol ester-treated cells was not significantly affected by supplemental LF provided at birth. Oral LF is absorbed in calves but does not alter PMN superoxide production and does not alter IgG absorption. PMID- 14765740 TI - The effects of oral magnesium hydroxide administration on rumen fluid in cattle. AB - This study was conducted to determine the effects of oral magnesium hydroxide administration on rumen fluid in cattle. Six lactating Holstein cows (4-7 years of age) with rumen fistulas were studied. Cattle were randomly assigned to receive boluses of magnesium hydroxide (162 g) or a powdered form (450 g dissolved in 3.5 L of water) PO daily for 3 days. Analysis of rumen fluid, blood gas tensions, and pH and measurement of serum magnesium concentrations were conducted daily. The study was discontinued after 72 hours, or sooner if rumen pH exceeded 8.0. After at least 3 weeks, the study was repeated with each cow receiving the other form of magnesium hydroxide (powder or bolus). Compared with baseline rumen pH (mean +/- SD: 6.22 +/- 0.28), magnesium hydroxide boluses caused a significant increase (P < .05) in rumen pH after 48 (7.27 +/- 0.11) and 72 (8.01 +/- 0.16) hours of administration, whereas the powdered form caused a significant increase (P < .05) in rumen pH after 24 (7.54 +/- 0.19) and 48 (8.43 +/- 0.22) hours of administration. Both the powdered and bolus forms of magnesium hydroxide decreased rumen protozoal numbers and increased methylene blue reduction times compared with baseline values. There was no change in blood pH, bicarbonate, or base excess values. Serum magnesium concentrations were significantly increased (P < .05) in cows that received the magnesium hydroxide powder. The results of this study indicate that magnesium hydroxide has a potent alkalinizing effect on rumen pH and significantly decreases rumen microbial activity. PMID- 14765741 TI - Cystoscopic appearance of proliferative urethritis in 2 dogs before and after treatment. PMID- 14765742 TI - Loperamide toxicity in a collie with the MDR1 mutation associated with ivermectin sensitivity. PMID- 14765743 TI - Light-chain myeloma in a dog. PMID- 14765744 TI - Lymphangiosarcoma associated with primary lymphedema in a Bouvier des Flandres. PMID- 14765745 TI - Sinonasal adenocarcinoma causing central nervous system disease in a horse. PMID- 14765746 TI - Acute respiratory distress syndrome and fatal interstitial pneumonia associated with equine influenza in a neonatal foal. PMID- 14765747 TI - Evaluation of a p30 gene-based real-time reverse transcriptase polymerase chain reaction assay for detection of feline caliciviruses. AB - This report describes a feline calicivirus (FCV) p30 gene-based real-time SYBR Green I reverse transcriptase polymerase chain reaction (RT-PCR) assay that is capable of detecting low virus concentrations and a broad range of FCV isolates. The assay consisted of a 1-step RT-PCR reaction with primers delineating a 126 base-pair (bp) region of the FCV p30 gene. Sensitivity of the RT-PCR assay was determined to be equivalent to a FCV titer of 1.2 x 10(1) to 1.2 x 10(2) TCID50/mL. The assay was linear over a wide range of template concentrations and had a reaction efficiency of 95%. Specific FCV amplification products were detected from 51 wild-type FCV isolates, whereas specific products were not detected from a canine calicivirus, a rabbit calicivirus, and a bovine calicivirus. The primers used in this study amplified a large number of North American FCV isolates and further confirm the diagnostic utility of p30 gene based real-time RT-PCR for detection of FCV. PMID- 14765748 TI - Adjacent instability after instrumented lumbar fusion. AB - The invention of pedicle screw instrumentation has greatly improved outcomes of spinal fusion, which has become the treatment of choice for lumbar spondylolisthesis. As researchers accumulate experience, both theoretical and clinical advances are continually being reported. A review of the literature and the experience of the authors show that the development of adjacent instability, as in the breakdown of a neighboring unfixed motion segment, is a common consequence of an instrumented lumbar spine. This article reviews the risk factors and surgical treatment of adjacent instability. The authors believe that proper preoperative planning and complete surgical procedures are imperative to prevent adjacent instability. For those who need revision surgery, meticulous surgical techniques can achieve satisfactory results. PMID- 14765749 TI - Interstitial laser photocoagulation for treatment of benign prostatic hypertrophy: outcomes and cost effectiveness. AB - BACKGROUND: We examined the efficiency and cost effectiveness of a temperature feedback diode-laser system in the treatment of benign prostatic hypertrophy (BPH). METHODS: One hundred twenty patients with symptomatic BPH were included in this study between October 1997 and January 1998. Sixty of them were treated by transurethral resection of the prostate (TUR-P), and 60 patients were treated by temperature feedback interstitial laser coagulation (ILC). Direct and indirect cost parameters, such as operative time, operation-related consumables, duration of hospitalization, and amount of medication used were compared between the 2 groups. RESULTS: All subjective and objective urinary parameters exhibited significant improvement 12 months after ILC. A reduction of 26.8% (46.6 to 34.1 ml) of the pretreatment prostate volume was observed at 12 months following ILC. The duration of hospital stay, operative time, and postoperative medications were significantly lower for those receiving ILC (5.9 to 2.5 days, p < 0.001) than for those who underwent TUR-P. The variety of laboratory tests needed for preoperative evaluation was no less when ILC was chosen for treating BPH (p = 0.849). Indirect costs, such as investment in laser equipment and laser accessories were higher in the ILC group (p < 0.001). CONCLUSION: The low morbidity profile, particularly the absence of retrograde ejaculation, makes ILC a valuable and attractive option for treatment of BPH patients who wish to retain their ejaculation ability, who have serious underlying diseases, or who have surgical risks for TUR-P or other invasive modalities. PMID- 14765750 TI - Surgical treatment of oral verrucous carcinoma. AB - BACKGROUND: The aim of this study was to evaluate the outcome of patients with Verrucous carcinoma (VC) of oral cavity treated at the Chung Gung Memory Hospital with respect to the tumor control rates after surgery, the risk of lymph node metastasis and the role of radiation therapy. METHODS: Thirty-eight patients underwent primary treatment for VC of the oral cavity from January 1996 through February 2002. All of the patients had surgery as their primary treatment. In addition, all patients with sufficient details of the therapy and a minimum 1 year follow-up were selected for evaluation of survival and outcomes. RESULTS: In this study, 94.7% of patients were male and most of them had been exposed to betel nuts, cigarettes, and/or alcohol. The most common site was the buccal mucosa (57.9%), followed by the tongue (13.2%). T3 lesions were the most common type (34.2%). Only two patients had palpable cervical adenopathy during the initial evaluation. Twenty-five patients had free flap for reconstruction. The tumor control rate was 100%. At the time of analysis, no patient had suffered from recurrence in primary site or neck area. CONCLUSION: Surgical excision alone was effective for controlling VC, but elective neck dissection was not necessary even in patients in the advanced stages. PMID- 14765751 TI - Using a Y-shaped vein graft with drain-out branches to provide additional arterial sources for free flap reconstruction in injured lower extremities. AB - BACKGROUND: Reconstruction of the distal lower extremity with poor arterial circulation is a challenge for reconstructive surgeons, especially when a microsurgical free tissue transfer is necessary. One may face the problem of having no suitable recipient artery to be used except the main one. To overcome this difficult situation, utilization of a Y-shaped vein graft (YVG) was developed. METHODS: We report on 3 successful reconstructions of the lower extremities with free flaps using YVG to supply the arterial inflow and in which the distal circulation of the lower extremity was preserved at the same time. The therapeutic goals, design, surgical technique, and outcomes are clarified. The indications, advantages, and disadvantages are also discussed. RESULTS: All 3 patients achieved successful reconstruction of the affected limbs with uneventful surgery. Special microsurgical skills are unnecessary in this method. It can provide an additional arterial source, which is useful and reliable for revascularizing various kinds of free tissue. Utmost care must be taken to examine the direction of flow during harvesting since only the drainout branches can serve as an ideal YVG. CONCLUSION: We concluded that Y-shaped vein grafting is a good alternative technique to achieve free flap reconstruction of a distal lower extremity with simultaneous preservation of the distal circulation in selected difficult cases. PMID- 14765752 TI - The influence of the distance from the contact point to the crest of bone on the presence of the interproximal dental papilla. AB - BACKGROUND: Loss of the interproximal dental papilla may cause functional and, especially in the maxillary anterior region, phonetic and severe esthetic problems. The purpose of this study was to investigate whether the distance from the contact point to the bone crest on standardized periapical radiographs of the maxillary anterior teeth could be correlated with the presence of the interproximal papilla in Taiwanese patients. METHODS: In total, 200 interproximal sites of maxillary anterior teeth in 45 randomly selected patients were examined. Selected subjects were adult Taiwanese with fully erupted permanent dentition. The presence of the interproximal papilla was determined visually. If there was no visible space apical to the contact area, the papilla was recorded as being present. The distance from the contact point to the crest of bone was measured on standardized periapical radiographs using a paralleling technique with a RinnXCP holder. RESULTS: Data revealed that when the distance from the contact point to the bone crest on standardized periapical radiographs was 5 mm or less, the papillae were almost 100% present. When the distance was 6 mm, 51% of the papillae were present, and when the distance was 7 mm or greater, only 23% of the papillae were present. CONCLUSION: The distance from the contact point to the bone crest on standardized periapical radiographs of the maxillary anterior teeth is highly associated with the presence or absence of the interproximal papilla in Taiwanese patients, and is a useful guide for clinical evaluation. PMID- 14765753 TI - Progression of diabetic retinopathy after phacoemulsification in diabetic patients: a three-year analysis. AB - BACKGROUND: This retrospective study was conducted to analyze the progression rates of diabetic retinopathy (DR) after phacoemulsification in diabetic patients and to determine whether phacoemulsification causes the progression of DR. METHODS: The medical charts of diabetic patients who had undergone cataract surgery using phacoemulsification techniques during a 3-year period and had completed 3 years of follow-up were retrospectively reviewed. Data were collected to evaluate the visual outcomes and progression of DR postoperatively. The retinopathy progression rates of operated and nonoperated eyes were compared. RESULTS: Thirty-seven diabetic patients were enrolled. Binocular cataract surgeries were performed in 14 patients. Monocular cataract surgeries were performed in the remaining 23 patients whose nonoperated fellow eyes served as the control group. The postoperative retinopathy progression rates of eyes with preoperative mild to moderate nonproliferative diabetic retinopathy (NPDR) were significantly greater than eyes without preoperative retinopathy whether at postoperative 1 year or postoperative 3 years. No statistical significance was found for the rate of retinopathy progression between operated and nonoperated eyes at 1 year and 3 years postoperatively. Of 51 operated eyes, 33 (64.7%) achieved an uncorrected visual acuity of 20/40 at 1 year postoperatively, and 47 (92.1%) had improvement of visual acuity of more than two lines. CONCLUSION: Uncomplicated phocoemulsificaton had no significant influence on the postoperative progression of DR. Patients with preoperative NPDR had greater chances to show postoperative retinopathy progression. The majority of diabetic patients who had no DR or had mild to moderate NPDR achieved visual improvement after phacoemulsification. PMID- 14765754 TI - Surgical treatment of retinal detachment following acute retinal necrosis syndrome: surgical results in four patients. AB - BACKGROUND: Acute retinal necrosis (ARN) syndrome is an uncommon but severe ocular disease that typically affects otherwise healthy individuals. It is frequently complicated with retinal detachment and the visual prognosis in such patients is usually poor. METHODS: We operated on four eyes in four patients from 1999 through 2001. Three ophthalmologists in our hospital did these operations, respectively. The surgical methods included pars plana vitrectomy, lensectomy, encircling scleral buckling combined with membrane dissection, air-fluid exchange, endolaser photocoagulation, and retinal tamponade with silicone oil or perfluoropropane gas. RESULTS: Three patients received one operation and the other one needed a second operation to release the retinal traction. One patient needed a lensectomy at the time of vitrectomy. Macular attachment was achieved in all four eyes (100%). Vision improved in two patients but none achieved visual acuity better than 20/200. The complications were cataract in three patients, macular pucker in three, and silicone keratopathy in one. CONCLUSION: Our results suggest that modern vitrectomy techniques provide a very high retinal attachment rate in patients with retinal detachment following ARN syndrome. PMID- 14765755 TI - Endoscopic removal of a dental prosthesis in the hepatic flexure of the colon. AB - The diagnosis of impacted foreign body in the colon is usually delayed until the complications such as perforation or abscess formation occur. Here we describe a patient who presented with diffuse abdominal pain due to the impaction of a dental prosthesis in the hepatic flexure of the colon. The dental prosthesis, which was inadvertently swallowed, was successfully removed under colonoscopy. Unexplained abdominal pain should alert the clinician to the possibility of foreign body ingestion and further therapeutic colonoscopy may replace or lessen the need for surgical procedures to extract foreign bodies from the colon. PMID- 14765756 TI - Idiopathic granulomatous orbital inflammation. AB - Idiopathic granulomatous inflammation is regarded as a histopathologic variant of an orbital pseudotumor and has a similar clinical presentation and treatment. However, the differential diagnosis including sarcoidosis, Wegener's granulomatosis, trauma, and infection must be ruled out. We present a 77-year-old man with a biopsy-proven lacrimal gland granulomatous inflammation of an unknown origin. Management by surgical debulking via an anterior orbitotomy achieved a good result with minimal complications. PMID- 14765757 TI - Acute primary hematogenous osteomyelitis of the epiphysis: report of two cases. AB - Acute primary infection of the epiphysis is uncommon. We present 2 cases of acute osteomyelitis of the distal femoral epiphysis. They were not diagnosed until 10 days and 3 weeks, respectively, after the onset of symptoms. The epiphyseal infection spread into the knee joint, and surgical debridement was performed. The majority of reported cases in the medical literature are of bacterial etiology, and the most common pathogen is Staphylococcus aureus. We report a rare case which was infected by Salmonella enteritidis. Prompt diagnosis and early treatment are required to prevent further destruction and growth disturbance. PMID- 14765758 TI - Primary malignant melanoma of the nasal cavity. AB - Malignant melanoma is a highly lethal melanocytic neoplasm, usually affecting the skin. Primary malignant melanoma of the nasal cavity is rarely seen. Clinically, most patients display initial nonspecific symptoms of unilateral nasal obstruction or epistaxis. The prognosis is generally poor, with a mean survival time of 3.5 years. Extensive local invasion and distant metastasis to other organs may occur. The usual treatment of choice is radical excision. Radiotherapy and chemotherapy appear to have little effect. We report a fatal case of intranasal cavity malignant melanoma in which the patient initially presented with blood-tinged sputum, productive cough, and intermittent fever. Preoperative hepatic metastasis was found. Palliative surgery was performed to excise the nasal cavity tumor. Then, 6 courses of chemotherapy were further administered. Unfortunately, regional cervical nodal involvement and pancreatic head metastases occurred 1.5 years after the diagnosis. The patient's condition rapidly deteriorated, followed by death. We have chosen to discuss this aggressive condition because of its rarity and also to emphasize the importance of its early detection through vigilant attention to nonspecific nasal symptoms. A review of the literature concerning intranasal malignant melanoma is presented. We further discuss its possible etiology, site of origin, incidence, clinical presentations, principles of management, and outcome. PMID- 14765759 TI - Correlates of college student gambling in the United States. AB - The authors report findings from the first national survey of gambling among college students. They collected information from 10,765 students attending 119 scientifically selected colleges included in the 2001 Harvard School of Public Health College Alcohol Study (CAS), which indicated that 42% of responding students gambled in the last school year and 2.6% gambled weekly or more frequently. These findings contradict the widely held opinion based on previous studies that gambling is prevalent among college students. Previous studies used convenience samples that might have overestimated the risk for gambling problems. Male students and students of legal age (> or = 21 y) were more likely than others to gamble. Availability of gambling, measured by the number of types of gambling venues located in the schools' states, influenced their decisions to gamble. The concordance of characteristics associated with both gambling and binge drinking suggested a disposition to take risks that is independent of the risk objects. PMID- 14765760 TI - Risk factors for Chlamydia trachomatis infection in a California collegiate population. AB - Chlamydia trachomatis infection in college students has not undergone a detailed large-scale evaluation. The authors undertook a cross-sectional study of 4,086 students enrolled on the campuses of California State University, Sacramento, and 3 local community colleges from fall 2000 through spring 2002. They used an outreach screening model to determine the presence of infection by testing urine for chlamydial DNA by polymerase chain reaction. Participants completed demographic and behavioral risk questionnaires and a chlamydia knowledge assessment. Overall, 3.4% of participants tested positive for infection. Screening of male and female students was evenly distributed and incidence rates for males (3.03%) and females (3.78%) were similar Variables associated with significantly increased risk were younger than 25 years, ethnicity other than White, more than one sexual partner in the preceding year or a new partner in the preceding 2 months, and current symptoms. Innovative outreach programs to screen a majority of at-risk college students should be developed. PMID- 14765761 TI - Binge drinking during the first semester of college: continuation and desistance from high school patterns. AB - Students' first semester on campus may set the stage for their alcohol use/misuse throughout college. The authors surveyed 274 randomly sampled first-semester freshmen at a large southwestern university on their past 2 weeks' binge drinking, their high school binge drinking, and psychosocial factors possibly associated with drinking. They conducted separate analyses among high school nonbinge drinkers (testing for predictors of college binge onset vs continued nonbinge drinking) and high school binge drinkers (testing for predictors of continued binge drinking in college vs desistance from drinking). In both analyses, the variables that predicted college binge drinking largely revolved around gregarious socializing (e.g, partying, having a social network of individuals who drank relatively heavily). Gender was predictive only among high school nonbinge drinkers; women had a higher probability than did men of adopting binge drinking in college. PMID- 14765762 TI - Assessing overweight, obesity, diet, and physical activity in college students. AB - The authors surveyed 738 college students aged 18 to 27 years to assess overweight, obesity, dietary habits, and physical activity. They used BMI (body mass index) > or = 25 kg/m2 or BMI > or = 85th percentile and BMI > or = 30 kg/m2 or BMI > or = 95th percentile to estimate overweight and obesity in those aged < or = 19 years. To define overweight and obesity in those > or = 20 years, they used BMI > or = 25 kg/m2 and > or = 30 kg/m2. They found overweight rates of 21.6% using BMI directly and 16.2% using BMI percentile and obesity rates of 4.9% using BMI directly and 4.2% using BMI percentile. More than 69% of the participants reported < 5 servings of fruits and vegetables per day and more than 67% reported < 20 g of fiber per day; participants reported physical activity on fewer than 3 d/wk. Most college students are not meeting dietary and physical activity guidelines, suggesting the need for prevention interventions and increased understanding of overweight in college students. PMID- 14765763 TI - Providing medical information to college health center personnel: a circuit Librarian Service at the University of Illinois. AB - College health center personnel are no different from other health practitioners in their need for medical information. To help meet this need, the McKinley Health Center, University of Illinois Urbana-Champaign, developed a partnership in 1997 with the Library of the Health Sciences-Urbana, a regional site library of the University of Illinois at Chicago campus. This partnership led to the adoption of circuit librarianship, a dynamic outreach model, to enhance access to health information for McKinley Health Center personnel. A circuit librarian consults with health center personnel during regularly scheduled on-site visits to the center and its satellite office. Upon returning to the resource library, the circuit librarian conducts research for clinical information on behalf of the center's personnel, then sees that articles, books, and relevant Web sites are identified and delivered to assist in answering questions regarding disease management, drug therapy, wellness, and health administration. PMID- 14765764 TI - Changing the focus of college alcohol prevention programs. PMID- 14765765 TI - Use of metformin is a cause of vitamin B12 deficiency. PMID- 14765766 TI - Preventive therapy for diabetes: lifestyle changes and the primary care physician. PMID- 14765767 TI - Jaundice in the adult patient. AB - Jaundice in an adult patient can be caused by a wide variety of benign or life threatening disorders. Organizing the differential diagnosis by prehepatic, intrahepatic, and posthepatic causes may help make the work-up more manageable. Prehepatic causes of jaundice include hemolysis and hematoma resorption, which lead to elevated levels of unconjugated (indirect) bilirubin. Intrahepatic disorders can lead to unconjugated or conjugated hyperbilirubinemia. The conjugated (direct) bilirubin level is often elevated by alcohol, infectious hepatitis, drug reactions, and autoimmune disorders. Posthepatic disorders also can cause conjugated hyperbilirubinemia. Gallstone formation is the most common and benign posthepatic process that causes jaundice; however, the differential diagnosis also includes serious conditions such as biliary tract infection, pancreatitis, and malignancies. The laboratory work-up should begin with a urine test for bilirubin, which indicates that conjugated hyperbilirubinemia is present. If the complete blood count and initial tests for liver function and infectious hepatitis are unrevealing, the work-up typically proceeds to abdominal imaging by ultrasonography or computed tomographic scanning. In a few instances, more invasive procedures such as cholangiography or liver biopsy may be needed to arrive at a diagnosis. PMID- 14765768 TI - Facilitating treatment adherence with lifestyle changes in diabetes. AB - Healthy eating and increased physical activity can prevent or delay diabetes and its complications. Techniques that facilitate adherence to these lifestyle changes can be adapted to primary care. Often, the patient's readiness to work toward change must be developed gradually. To prepare patients who are reluctant to change, it is effective to assess and address their conviction and confidence. Patients facing the long-term task of making lifestyle changes benefit from assistance in setting highly specific behavior-outcome goals and short-term behavior targets. Individualization is achieved by tailoring these goals and targets to the patient's preferences and progress, building the patient's confidence in small steps, and implementing more intensive interventions according to a stepped-care model. At each office visit, physician follow-up of the patient's self-monitored goals and targets enhances motivation and allows further customization of the plan. A coaching approach can be used to encourage positive choices, develop self-sufficiency, and assist the patient in identifying and overcoming barriers. More intensive intervention using a team approach maximizes adherence. PMID- 14765769 TI - Information from your family doctor. Sticking to lifestyle changes: increasing physical activity. PMID- 14765770 TI - Information from your family doctor. Sticking to lifestyle changes: healthy eating. PMID- 14765771 TI - Treating acute bronchiolitis associated with RSV. AB - Treatment for infants with bronchiolitis caused by respiratory syncytial virus (RSV) includes supplemental oxygen, nasal suctioning, fluids to prevent dehydration, and other supportive therapies. High-risk children who should be hospitalized include those younger than three months and those with a preterm birth, cardiopulmonary disease, immunodeficiency, respiratory distress, or inadequate oxygenation. Inhaled beta2-agonist bronchodilators, the anticholinergic agent ipratropium bromide, and nebulized epinephrine have not been shown to be effective for treating RSV bronchiolitis. However, the Agency for Healthcare Research and Quality states that nebulized epinephrine and nebulized ipratropium bromide are possibly effective. The appropriate use of corticosteroids remains controversial. They may provide some benefit but meta analyses of clinical trial results are inconsistent. Prophylaxis with RSV intravenous immune globulin or palivizumab, a human monoclonal antibody, can reduce hospitalization rates in high-risk patients, although difficulties with administering the medications and high costs may preclude their widespread use. The use of common infection-control measures can reduce nosocomial transmission of RSV infections. PMID- 14765772 TI - Hyperparathyroidism. AB - Primary hyperparathyroidism is the most frequent cause of hypercalcemia in ambulatory patients. The condition is most common in postmenopausal women, although it can occur in persons of all ages, including pregnant women. If symptoms are present, they are attributable to hypercalcemia and may include weakness, easy fatigability, anorexia, or anxiety. However, most persons have no symptoms, and primary hyperparathyroidism usually is diagnosed after an elevated serum calcium level is found incidentally on multiphasic chemistry panel testing. Persistent hypercalcemia and an elevated serum parathyroid hormone level are the diagnostic criteria for primary hyperparathyroidism. Other causes of hypercalcemia are rare, and usually are associated with low (or sometimes normal) parathyroid hormone levels. Malignancy is the most frequent cause of hypercalcemia in hospitalized patients. Parathyroidectomy is the definitive treatment for primary hyperparathyroidism. When performed by experienced endocrine surgeons, the procedure has success rates of 90 to 95 percent and a low rate of complications. Asymptomatic patients who decline surgery and meet criteria for medical management must commit to conscientious long-term monitoring. Any unexplained elevation of the serum calcium level should be evaluated promptly to prevent complications from hypercalcemia. PMID- 14765773 TI - Information from your family doctor. Hyperparathyroidism. PMID- 14765775 TI - Information from your family doctor. Head lice. PMID- 14765774 TI - Pediculosis and scabies. AB - Pediculosis and scabies are caused by ectoparasites; patients usually present with itching. Head and pubic lice infestations are diagnosed by the visualization of insects or viable nits (eggs). Primary treatment is topically administered 1 percent permethrin. Malathion is one alternative for treatment failures. The importance of environmental measures to prevent infestation is a matter of controversy. Pubic lice are treated the same as head lice, but this finding should prompt evaluation for other sexually transmitted diseases. Body lice infestation should be suspected when symptoms of generalized itching occur in persons who do not change or wash their clothing or bedding regularly; lice may be found in the seams of their clothing. Topically administered permethrin may help to eradicate body lice, but personal hygiene measures are essential for successful treatment. Classic scabies in adults can be recognized by a pruritic, papular rash with excoriations; in infants, small children, and the immunocompromised, the rash may include vesicles, pustules, or nodules. Primary treatment for scabies is permethrin cream and environmental measures are important to prevent recurrent infestation. Generalized crusted scabies is best treated with oral ivermectin. PMID- 14765776 TI - Behavioral interventions to promote breastfeeding: recommendations and rationale. PMID- 14765777 TI - Does a low-sodium diet reduce blood pressure? PMID- 14765778 TI - Tegaserod (Zelnorm) for irritable bowel syndrome. PMID- 14765779 TI - Suspected pulmonary embolism: part I. Evidence-based clinical assessment. PMID- 14765781 TI - Information from your family doctor. Antibiotics: when they can and can't help. PMID- 14765780 TI - Velvety axillary lesions. PMID- 14765782 TI - Information from your family doctor. Influenza vaccine. PMID- 14765783 TI - Information from your family doctor. Flu and colds. PMID- 14765784 TI - Information from your family doctor. Acute bronchitis. PMID- 14765785 TI - Information from your family doctor. Sore throat. PMID- 14765786 TI - ACOG releases bulletin on managing cervical insufficiency. PMID- 14765787 TI - Washington state dairy cow nation's first case of BSE. PMID- 14765788 TI - USAHA supports livestock identification plan, bolstering security for food supply. PMID- 14765789 TI - Army veterinarian is militarys man with the plan. PMID- 14765790 TI - Introducing a valuable new benefit: the Health Savings Account. PMID- 14765791 TI - Superstar stamp sells out. PMID- 14765792 TI - Wishes to clarify misleading anesthesia myth. PMID- 14765793 TI - Questions JAVMA's method of reviewing books. PMID- 14765794 TI - What is your diagnosis? Multifocal nonconsolidating alveolar pattern with mineralized foci in multiple ling fields and moderate pleural effusion. PMID- 14765795 TI - ECG of the month. Supraventricular tachycardia with second-degree AV block Mobitz type I. PMID- 14765796 TI - Long-term risks and benefits of early-age gonadectomy in cats. AB - OBJECTIVE: To evaluate the long-term risks and benefits of early-age gonadectomy, compared with traditional-age gonadectomy, among cats adopted from a large animal shelter. DESIGN: Retrospective cohort study. ANIMALS: 1,660 cats. PROCEDURE: Cats underwent gonadectomy and were adopted from an animal shelter before 1 year of age; follow-up was available for as long as 11 years after surgery (median follow up time, 3.9 years). Adopters completed a questionnaire about their cats' behavior and medical history. When possible, the cats' veterinary records were reviewed. Statistical analyses were conducted to identify any associations between the occurrence of 47 medical and behavioral conditions and the cats' age at gonadectomy. RESULTS: Among male cats that underwent early-age gonadectomy (< 5.5 months of age), the occurrence of abscesses, aggression toward veterinarians, sexual behaviors, and urine spraying was decreased, whereas hiding was increased, compared with cats that underwent gonadectomy at an older age. Among male and female cats that underwent early-age gonadectomy, asthma, gingivitis, and hyperactivity were decreased, whereas shyness was increased. CONCLUSIONS AND CLINICAL RELEVANCE: Gonadectomy before 5.5 months of age was not associated with increased rates of death or relinquishment or occurrence of any serious medical or behavioral condition and may provide certain important long-term benefits, especially for male cats. Animal shelters can safely gonadectomize cats at a young age, and veterinarians should consider recommending routine gonadectomy for client-owned cats before the traditional age of 6 to 8 months. PMID- 14765797 TI - Long-term risks and benefits of early-age gonadectomy in dogs. AB - OBJECTIVE: To evaluate the long-term risks and benefits of early-age gonadectomy, compared with traditional-age gonadectomy, among dogs adopted from a large animal shelter. DESIGN: Retrospective cohort study. ANIMALS: 1,842 dogs. PROCEDURE: Dogs underwent gonadectomy and were adopted from an animal shelter before 1 year of age; follow-up was available for as long as 11 years after surgery. Adopters completed a questionnaire about their dogs' behavior and medical history. When possible, the dogs' veterinary records were reviewed. Associations between the occurrence of 56 medical and behavioral conditions and dogs' age at gonadectomy were evaluated. RESULTS: Among female dogs, early-age gonadectomy was associated with increased rate of cystitis and decreasing age at gonadectomy was associated with increased rate of urinary incontinence. Among male and female dogs with early-age gonadectomy, hip dysplasia, noise phobias, and sexual behaviors were increased, whereas obesity, separation anxiety, escaping behaviors, inappropriate elimination when frightened, and relinquishment for any reason were decreased. CONCLUSIONS AND CLINICAL RELEVANCE: Because early-age gonadectomy appears to offer more benefits than risks for male dogs, animal shelters can safely gonadectomize male dogs at a young age and veterinary practitioners should consider recommending routine gonadectomy for client-owned male dogs before the traditional age of 6 to 8 months. For female dogs, however, increased urinary incontinence suggests that delaying gonadectomy until at least 3 months of age may be beneficial. PMID- 14765798 TI - Evaluation of cisplatin combined with piroxicam for the treatment of oral malignant melanoma and oral squamous cell carcinoma in dogs. AB - OBJECTIVE: To determine the maximum tolerated dose (MTD) of cisplatin administered with piroxicam, the antitumor activity and toxicity of cisplatin combined with piroxicam in dogs with oral malignant melanoma (OMM) and oral squamous cell carcinoma (SCC), and the effects of piroxicam on the pharmacokinetics of cisplatin in dogs with tumors. DESIGN: Prospective nonrandomized clinical trial. ANIMALS: 25 dogs. PROCEDURE: Dogs were treated with a combination of cisplatin (escalating dose with 6 hours of diuresis with saline [0.9% NaCI] solution) and piroxicam (0.3 mg/kg 10.14 mg/lb], PO, q 24 h). The initial cisplatin dose (50 mg/m2) was increased by 5 mg/m2 until the MTD was reached. Tumor stage and size were determined at 6-week intervals during treatment. The pharmacokinetics of cisplatin were determined in dogs receiving a combination of cisplatin and piroxicam during the clinical trial and dogs that were treated with cisplatin alone. RESULTS: 11 dogs with OMM and 9 dogs with SCC were included in the clinical trial. The MTD of cisplatin when administered in combination with piroxicam was 50 mg/m2. Tumor remission occurred in 5 of 9 dogs with SCC and 2 of 11 dogs with OMM. The most common abnormality observed was renal toxicosis. Clearance of cisplatin in dogs that were treated with cisplatin alone was not significantly different from that in dogs treated with a combination of cisplatin and piroxicam. CONCLUSIONS AND CLINICAL RELEVANCE: Cisplatin administered in combination with piroxicam had antitumor activity against OMM and SCC. The level of toxicity was acceptable, although renal function must be monitored carefully. PMID- 14765799 TI - Ultrasonographic assessment of hemodynamic changes in the portal vein during surgical attenuation of congenital extrahepatic portosystemic shunts in dogs. AB - OBJECTIVE: To determine portal hemodynamic changes associated with surgical shunt ligation and establish ultrasonographic criteria for determining the optimal degree of shunt narrowing and predicting outcome. DESIGN: Case series. ANIMALS: 17 dogs, each with a single congenital extrahepatic portosystemic shunt. PROCEDURE: Pre- and postligation flow velocities and flow directions were determined by Doppler ultrasonography intraoperatively in the shunt and in the portal vein cranial and caudal to the shunt origin. Outcome was evaluated 1 month after surgery by measuring blood ammonia concentration and performing abdominal ultrasonography. RESULTS: Hepatofugal flow was detected in 9 of 17 dogs before shunt attenuation in the portal segment that was between the shunt origin and the entering point of the gastroduodenal vein. If hepatofugal flow became hepatopetal after shunt ligation, hyperammonemia resolved. Hepatofugal portal flow was caused by blood that flowed from the gastroduodenal vein toward the shunt. Shunt attenuation converted hepatofugal flow to hepatopetal in the shunt in 12 of 17 dogs. Chronic portal hypertension developed or perioperative death occurred when the portal congestion index caudal to the shunt origin increased by > 3.6 times. CONCLUSIONS AND CLINICAL RELEVANCE: After hepatopetal flow in the cranial portal vein and the shunt is established, further shunt narrowing is contraindicated. Increase of the portal congestion index caudal to the shunt > 3.5 times should be avoided. Poor outcome because of severe hypoplasia of the portal branches can be expected if the flow direction remains hepatofugal after shunt occlusion cranial to the shunt origin. PMID- 14765800 TI - Concurrence between clinical and pathologic diagnoses in a veterinary medical teaching hospital: 623 cases (1989 and 1999). AB - OBJECTIVE: To determine whether there was a decline in the percentage of dogs undergoing necropsies and whether there was substantial agreement or disagreement between clinical and pathologic diagnoses. DESIGN: Retrospective study. ANIMALS: 623 dogs. PROCEDURE: Medical records of hospitalized dogs that died or were euthanatized and necropsied at a veterinary teaching hospital in 1989 and 1999 were reviewed. Clinical and pathologic diagnoses were recorded and compared. RESULTS: There was a significant decline in the necropsy rate of hospitalized dogs that died or were euthanatized in 1999, compared with 1989. In both 1989 and 1999, there was disagreement between the clinical and pathologic diagnoses in approximately a third of the cases. CONCLUSIONS AND CLINICAL RELEVANCE: Despite improved diagnostic methods, the accuracy of diagnosis did not improve significantly in 1999, compared with 1989. Necropsy is the best method to assess overall diagnostic accuracy. Increased availability of teaching funds may promote efforts to have necropsies performed in veterinary teaching hospitals. PMID- 14765801 TI - Use of dapsone in the treatment of Pneumocystis carinii pneumonia in a foal. AB - A 6-month-old male Quarter Horse was evaluated for chronic respiratory tract disease. Diagnostic investigations revealed pulmonary inflammation; Pneumocystis carinii was detected within macrophages. Lymphocyte subpopulation phenotyping and immunoglobulin concentration analysis were performed and results suggested immune suppression. Trimethoprim-sulfamethoxazole administration was initiated; the colt was discharged but was reexamined 8 days later because of profuse diarrhea and endotoxemia. Bacterial culture of feces recovered Salmonella spp resistant to trimethoprim-sulfamethoxazole, and a diagnosis of antimicrobial-associated colitis was made. Bilateral fibrinous hypopyon developed and was treated with topical medication and intracameral injections of human recombinant tissue plasminogen activator. Dapsone (3 mg/kg [1.4 mg/lb], PO, q 24 h; dose extrapolated from human data) was administered for treatment of P carinii pneumonia (56-day treatment period). The colt recovered from the pneumonia and diarrhea. Dapsone may be a useful adjunct to traditional treatment for P carinii pneumonia in horses or as a sole medication for horses that cannot tolerate other treatments. PMID- 14765802 TI - Use of magnetic resonance imaging for identifying subchondral bone damage in horses: 11 cases (1999-2003). AB - OBJECTIVE: To assess the use of magnetic resonance (MR) imaging for identifying subchondral bone damage in the distal limbs of horses. DESIGN: Retrospective study. ANIMALS: 11 horses. PROCEDURE: Medical records of horses with lameness and subsequent evidence of subchondral bone damage as determined by MR imaging were reviewed. Severity and duration of lameness, results of diagnostic local anesthesia and diagnostic testing, surgical and necropsy findings, and treatment were recorded. Outcome was determined by follow-up information obtained from the owner or referring veterinarian. RESULTS: Lameness was localized by physical examination and diagnostic local anesthesia. Lameness was localized to the metacarpophalangeal or metatarsophalangeal joint in 4 horses, distal interphalangeal joint in 5 horses, and tarsocrural joint in 2 horses. The duration of lameness ranged from 2 weeks to 20 months. Magnetic resonance imaging of the affected joints revealed abnormal fluid accumulation within the subchondral bone. None of the abnormalities observed by MR imaging were detected by radiography. Subchondral bone damage was diagnosed in all horses. Arthroscopy of the affected joint was performed in 4 horses. Communication with the articular surface of the affected bone was suspected on the basis of results of MR imaging in 4 horses and was confirmed by arthroscopy in 1 horse and by necropsy in 1 horse. CONCLUSIONS AND CLINICAL RELEVANCE: Magnetic resonance imaging was useful for providing a diagnosis when other imaging techniques did not definitively identify the cause of lameness. Subchondral bone damage was clearly identified by MR imaging and should be considered as a cause of lameness in horses in which radiographic findings are unremarkable. PMID- 14765803 TI - Use of somatic cell counts and California mastitis test results from individual quarter milk samples to detect subclinical intramammary infection in dairy cattle from a herd with a high bulk tank somatic cell count. AB - OBJECTIVE: To determine whether somatic cell counts (SCCs) or California mastitis test (CMT) scores for individual quarter milk samples could be used to detect subclinical intramammary infection among dairy cattle in a herd with a high bulk tank SCC. DESIGN: Prospective clinical trial. ANIMALS: 278 Holstein-Friesian dairy cattle from a single herd. PROCEDURE: Individual quarter milk samples were collected and submitted for bacterial culture, California mastitis testing, and determination of SCC. Additional milk samples were collected 34 days later and submitted for bacterial culture. RESULTS: During the initial visit to the herd, milk samples were collected from all 278 cows. However, because of blind mammary quarters or missing data, results for 1,057 quarter milk samples were included. Bacterial culture did not yield any growth for 622 (58.8%) of these samples. Regardless of the cutoff that was used, sensitivity of the CMT score was < or = 0.50 and sensitivity of the SCC linear score (SCS) was < or = 0.60. For 497 mammary quarters, results of bacterial culture of samples collected 34 days apart were concordant; bacterial culture did not yield any growth for 342 (68.8%) of these quarters. Regardless of the cutoff that was used, sensitivity of the CMT score was < or = 0.61 and sensitivity of the SCS was < or = 0.76 for mammary quarters with concordant bacterial culture results. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that neither CMT score nor SCC is sensitive enough to be useful as a screening test for identifying infected mammary quarters among dairy cattle in a herd with high bulk tank SCC. PMID- 14765804 TI - Ice cream structural elements that affect melting rate and hardness. AB - Statistical models were developed to reveal which structural elements of ice cream affect melting rate and hardness. Ice creams were frozen in a batch freezer with three types of sweetener, three levels of the emulsifier polysorbate 80, and two different draw temperatures to produce ice creams with a range of microstructures. Ice cream mixes were analyzed for viscosity, and finished ice creams were analyzed for air cell and ice crystal size, overrun, and fat destabilization. The ice phase volume of each ice cream were calculated based on the freezing point of the mix. Melting rate and hardness of each hardened ice cream was measured and correlated with the structural attributes by using analysis of variance and multiple linear regression. Fat destabilization, ice crystal size, and the consistency coefficient of the mix were found to affect the melting rate of ice cream, whereas hardness was influenced by ice phase volume, ice crystal size, overrun, fat destabilization, and the rheological properties of the mix. PMID- 14765805 TI - Preference mapping of Cheddar cheese with varying maturity levels. AB - The objective of this study was to evaluate the flavor profile and acceptability of 7 Cheddar cheeses of varying maturity using descriptive analysis and consumer acceptance tests. Cheddar cheeses (n = 7) ranging in age from 1 to 19 mo were selected based on age, geographic region, and flavor profile. Descriptive sensory profiles of selected cheeses were determined using a trained panel (n = 14) and an established cheese flavor sensory language. Cheeses were evaluated for consumer acceptability in two demographic locations: North Carolina and Oregon. Consumers (n = 100 at each location) assessed the cheeses for overall liking and other consumer attributes. Cheddar cheeses demonstrated distinct differences in descriptive sensory profiles. Average consumer responses between the two locations were not different. Six distinct consumer clusters were identified, and the number of consumers in these clusters differed between the two locations. Consumers differentiated "young" and "aged" cheese flavor, but both young and mature cheeses were perceived by consumers as exhibiting intense Cheddar cheese flavors. Cheddar cheese acceptance varies widely among consumers and is related to consumer preferences for distinct cheese flavor profiles. PMID- 14765806 TI - Effects of milk powders in milk chocolate. AB - The physical characteristics of milk powders used in chocolate can have significant impact on the processing conditions needed to make that chocolate and the physical and organoleptic properties of the finished product. Four milk powders with different particle characteristics (size, shape, density) and "free" milk fat levels (easily extracted with organic solvent) were evaluated for their effect on the processing conditions and characteristics of chocolates in which they were used. Many aspects of chocolate manufacture and storage (tempering conditions, melt rheology, hardness, bloom stability) were dependent on the level of free milk fat in the milk powder. However, particle characteristics of the milk powder also influenced the physical and sensory properties of the final products. PMID- 14765807 TI - Changes in vitamin C concentrations in plasma and milk from dairy cows after an intramammary infusion of Escherichia coli. AB - Plasma and milk concentrations of ascorbic acid and dehydro-L-ascorbic acid (DHAA) were measured before and after 21 Holstein cows (approximately 26 DIM) were given an intramammary infusion of Escherichia coli. Blood, milk from the unchallenged quarters, and milk from the challenged gland were sampled immediately before challenge (d 0) and 24 h and 7 d postchallenge. Plasma vitamin C (ascorbic acid + DHAA) concentrations decreased 39%, and concentrations of vitamin C and ascorbic acid in milk from the challenged quarter decreased 52 and 62%, respectively, in samples taken 24 h postchallenge. No change was observed in vitamin C concentrations in milk from unchallenged quarters. The concentration of DHAA in milk from challenged quarters increased 67% 24 h postchallenge. The duration of clinical mastitis, peak body temperature, number of colony-forming units of E. coli isolated from the infected gland, and loss in milk yield were associated with a change in concentration of vitamin C in milk from the challenged quarter. Increased severity of clinical signs was associated with large decreases in concentration of vitamin C in milk from the challenged quarter. Similar, but statistically weaker, relationships were observed for changes in plasma vitamin C concentrations. PMID- 14765808 TI - Identification of Corynebacterium bovis by endonuclease restriction analysis of the 16S rRNA gene sequence. AB - Despite its high prevalence within the bovine mammary gland, Corynebacterium bovis is considered a minor pathogen and of limited clinical significance. It has been suggested that intramammary infection with C. bovis may protect quarters against subsequent infection with other pathogens. The literature has produced much conflicting data on the subject. A possible explanation for some of the divergence of opinion on the subject is incorrect identification of isolates in previous studies. This paper describes a novel method for differentiating C. bovis from other lipophilic Corynebacterium species based on endonuclease restriction analysis. The 16S rRNA gene sequences for all known lipophilic Corynebacterium species were obtained from published data and analyzed. It was predicted that endonuclease restriction with HindIII and SmaI could be used to differentiate C. bovis from all other known lipophilic Corynebacterium species. The method was successfully employed to identify 741 of 762 (97.2%) lipophilic Corynebacterium species as C. bovis. Twenty one (2.8%) were identified as species other than C. bovis. Using this technique, it was demonstrated that it is not safe to assume that all lipophilic coryneform organisms isolated from bovine milk samples are C. bovis. This method is an alternative to more traditional methods of identification in large scale studies until methods such as 16S rRNA gene sequencing become more widely available. PMID- 14765809 TI - Changes in milk composition as affected by subclinical mastitis in sheep. AB - The mechanism of the effects of glandular-level subclinical mastitis in dairy sheep on milk yield and on its composition as expressed in curd yield was studied. Thirty-six Israeli-Assaf dairy sheep with one udder half infected with identified coagulase-negative staphylococci and the contralateral gland free of bacteria were chosen. The milk yield of the infected halves was significantly lower than that of the uninfected ones (0.36 vs. 0.76 kg/milking). The somatic cell count and N-acetyl-beta-D-glucosaminidase activity were significantly higher in the infected halves than in the uninfected ones. The plasminogen activator and plasmin (PL) activities were significantly higher in the infected glands than in the uninfected ones, whereas plasminogen (PLG) activity and the ratio PLG:PL were significantly lower in the infected glands. Concentrations of Ca2+ did not differ, whereas Ca2+ activity was significantly lower and proteose peptone concentration was 2.4 times as high in the infected glands than in the uninfected ones. Curd yield was significantly lower in the infected glands than in the uninfected ones. PMID- 14765810 TI - Differences in heat tolerance between preimplantation embryos from Brahman, Romosinuano, and Angus breeds. AB - Exposure to 41 degrees C reduces development of embryos of heat-sensitive breeds (Holstein and Angus) more than for embryos of the heat-tolerant Brahman breed. Here it was tested whether embryonic resistance to heat shock occurs for a thermotolerant breed of different genetic origin than the Brahman. In particular, the thermal sensitivity of in vitro produced embryos of the Romosinuano, a Bos taurus, Criollo-derived breed, was compared to that for in vitro produced Brahman and Angus embryos. At d 4 after insemination, embryos > or = 8 cells were randomly assigned to control (38.5 degrees C) or heat shock (41 degrees C for 6 h) treatments. Heat shock reduced the proportion of embryos that developed to the blastocyst stage on d 8 after insemination. At 38.5 degrees C, there were no significant differences in development between breeds. Among embryos exposed to 41 degrees C, however, development was lower for Angus embryos than for Brahman and Romosinuano embryos. Furthermore, an Angus vs. (Brahman + Romosinuano) x temperature interaction occurred because heat shock reduced development more in Angus (30.3 +/- 4.6% at 38.5 degrees C vs. 4.9 +/- 4.6% at 41 degrees C) than in Brahman (25.1 +/- 4.6% vs. 13.6 +/- 4.6%) and Romosinuano (28.3 +/- 4.1% vs. 17.5 +/- 4.1%). Results demonstrate that embryos from Brahman and Romosinuano breeds are more resistant to elevated temperature than embryos from Angus. Thus, the process of adaptation of Brahman and Romosinuano breeds to hot environments resulted in both cases in selection of genes controlling thermotolerance at the cellular level. PMID- 14765811 TI - Comparison of techniques for measurement of rumen pH in lactating dairy cows. AB - Subacute rumen acidosis is thought to be a common condition in early lactating dairy cattle; however, diagnosis is difficult. There are currently only two techniques available for measuring rumen pH under field conditions: rumenocentesis and oral stomach tube. Sixteen rumen-fistulated cows were sampled in four sites of the rumen (cranial-ventral, caudal-ventral, central, and cranial dorsal) with a rumen cannula. Rumen pH results were compared to those obtained at the same time with rumenocentesis and with an oro-ruminal (Geishauser) probe. Rumen fluid was obtained between 6 and 12 wk of lactation. Samples were analyzed for pH, lactate, bicarbonate, sodium, potassium, and chloride. Rumen pH results were also compared to those obtained from 24-h continuous rumen pH measurement using indwelling rumen pH probes. Oro-ruminal probe samples had the highest pH values and the highest bicarbonate concentrations. Rumenocentesis samples had the lowest pH values and the lowest bicarbonate concentrations. Small differences in electrolyte concentrations were noted among rumen fluid collection techniques in the different rumen sites. The highest correlations of rumen pH were obtained between rumenocentesis and rumen cannulation (cranial-ventral), and between rumen cannulation (cranial-ventral) and the 24-h indwelling pH meter. Compared with samples obtained from the cranial-ventral rumen, rumenocentesis was more sensitive than the oro-ruminal probe in the measurement of low rumen pH; both techniques were moderately specific. The most accurate field technique was rumenocentesis. Improved field techniques are required for better on-farm diagnosis of subacute rumen acidosis. PMID- 14765812 TI - Effects of conjugated linoleic acid isomers on lipid metabolism and gluconeogenesis in monolayer cultures of bovine hepatocytes. AB - The objective was to determine the effects of linoleic acid and different isomers of conjugated linoleic acid (CLA) at different concentrations on hepatic lipid and glucose metabolism in the bovine. Monolayer cultures of hepatocytes obtained from 7- to 10-d-old Holstein bull calves were exposed to treatments from 16 to 64 h after plating. The treatments included 1.0 mM palmitic acid plus either 0.1 or 1.0 mM of cis-9, cis-12 linoleic acid, cis-9, trans-11 CLA, or trans-10, cis-12 CLA. Metabolism of palmitic acid to cellular triacylglycerol (TAG) was decreased when media contained cis-9, trans-11 compared with trans-10, cis-12 CLA. Total cellular TAG content was increased for the CLA isomers compared to cis-9, cis-12 linoleic acid. Both CLA isomers increased palmitic acid incorporation into phospholipids, cholesterol, and media triacylglycerol compared with cis-9, cis-12 linoleic acid at a concentration of 1.0 mM. Increasing the concentration of treatment fatty acids from 0.1 to 1.0 mM decreased oxidation of palmitic acid to acid-soluble products, but no effects of fatty acids were observed. There was no treatment effect on rates of gluconeogenesis from propionic acid. Overall, CLA isomers elicited changes in palmitic acid metabolism to cellular and media triacylglycerol, and cellular phospholipids and cholesterol, but had little or no effect on other measured pathways of lipid metabolism or gluconeogenesis in bovine hepatocytes. PMID- 14765813 TI - Influence of transition diets on occurrence of subclinical laminitis in Holstein dairy cows. AB - Pre- and postpartum diets varying in energy and fiber were studied for effects on subclinical laminitis in Holstein cows. Also, ruminal acidosis was examined relative to sole hemorrhages and ulcers. Cows (n = 98) were assigned to a 2 x 2 factorial arrangement of treatments in a randomized complete block. Diets high in net energy of lactation (NE(L)) and low fiber were classified as high NE(L), whereas low NE(L) and higher fiber were defined as low NE(L). Two diets were fed for 3 wk before calving and 2 others fed for 3 wk postpartum resulting in 4 treatment combinations: high NE(L), low NE(L); high NE(L), high NE(L); low NE(L), low NE(L); and low NE(L), high NE(L). Levels of NE(L) (Mcal/kg DM), percentage of acid detergent fiber, and percentage of neutral detergent fiber for low NE(L) vs. high NE(L) prepartum diets, were 1.51, 30.2, 47.2 vs. 1.65, 23.4, 39.8, respectively, and 1.70, 22.4, 36.8 vs. 1.77, 17.5, 31.4 for low NE(L) vs. high NE(L) lactating diets, respectively. A single diet was fed after 21 d in milk (DIM). Measures of hoof discoloration, hemorrhage, and/or ulcer formation were done at about 45 d before calving, and near 28 and 70 DIM. Rumenocentesis was performed 14 d before calving and at 8, 22, and 70 DIM. Hoof scores among treatments were similar at 28 DIM. The low NE(L), high NE(L) group had less desirable hoof scores than high NE(L), high NE(L), or low NE(L), low NE(L) groups from 55 to 75 DIM. Rumen pH did not differ by treatment before calving. The lowest postpartum rumen pH was significantly lower and rates of ruminal acidosis (pH < or = 5.8) at 8 and 22 DIM were higher for groups fed high NE(L) after calving. No direct relationship between low postpartum rumen pH and hoof scores on individual cows was found. Low NE(L) before calving and high NE(L) right after calving may increase risk of subclinical laminitis if not carefully managed. PMID- 14765814 TI - Comparison of timed AI after synchronized ovulation to AI at estrus: reproductive and economic considerations. AB - A timed artificial insemination (TAI) protocol using OvSynch was compared to artificial insemination (AI) at detected estrus in 2 large dairy herds differing in reproductive management. Cows were synchronized for TAI starting at 62 and 42 d in milk in herds 1 and 2, respectively. The OvSynch regimen included: GnRH (buserelin) at 0.02 mg (i.m.) on d 0; PGF2alpha (tiaprost) at 0.75 mg (i.m.) on d 7; buserelin at 0.02 mg (i.m.) on d 9; and TAI 16 to 20 h later. After TAI, cows seen in estrus received AI, whereas cows diagnosed not pregnant were resynchronized for TAI. Control cows received AI based on detected estrus after voluntary waiting periods of 72 d in herd 1 and 50 d in herd 2. An economic analysis included costs associated with days open, culling, AI, synchrony products, treatment, and examinations. A sensitivity analysis of those variables determined effects on total costs per pregnancy. Use of OvSynch reduced intervals to first AI and days open in both herds and reduced culling for infertility in herd 2. Conception rates for first AI at detected estrus were significantly higher compared to TAI in both herds and for overall AI at estrus in herd 2. For groups assigned to AI at estrus, mean 21-d submission rates over 200 d for AI were higher in herd 1 than in herd 2 (55.6 vs. 28.6%). Days open and culling were the major cost factors. Although OvSynch improved reproduction in both herds, AI based on detected estrus was economically superior in herd 1, whereas OvSynch was superior in herd 2. This was consistent across ranges of cost factors evaluated. Evaluation of synchrony protocols should include reproductive performance along with appropriate costs associated with treatments. Such costs may offset benefits to reproduction in herds with good estrous detection rates. PMID- 14765815 TI - Associations between pathogen-specific cases of clinical mastitis and somatic cell count patterns. AB - Associations were estimated between pathogen-specific cases of clinical mastitis (CM) and somatic cell count (SCC) patterns based on deviations from the typical curve for SCC during lactation and compared with associations between pathogen specific CM and lactation average SCC. Data from 274 Dutch herds recording CM over an 18-mo period were used. Pathogens found were Staphylococcus aureus, coagulase-negative staphylococci, Escherichia coli, Streptococcus dysgalactiae, Streptococcus uberis, streptococci other than Strep. dysgalactiae and Strep. uberis, and culture-negative samples. The dataset contained 245,595 test-day records on SCC, recorded in 24,012 lactations of 19,733 cows of different parities. Pattern definitions were based on three or five consecutive test-day records. The patterns differentiated between a short or longer period of increased SCC and also between lactations with and without recovery. Logistic regression was applied to identify associations between presence of patterns and occurrence of pathogens. Occurrence of overall CM in a lactation is equally or even more accurately predicted by the presence of SCC in that lactation, than by a lactation average SCC of more than 200,000 cells/mL. Patterns can also distinguish between chances of risk for specific mastitis-causing pathogens. Clinical E. coli mastitis was significantly associated with the presence of a short peak in SCC, whereas Staph. aureus was associated with long increased SCC. Streptococcus dysgalactiae was not strongly associated with any of the defined patterns of peaks in SCC, and no single unambiguous pattern was found for Strep. uberis. PMID- 14765816 TI - Characterization of a colostrum replacer and a colostrum supplement containing IgG concentrate and growth factors. AB - The objective of this study was to characterize absorption of colostrum replacer (CR) or supplement (CS) containing fractions of bovine plasma. Immunoglobulin concentrate (IGC) was prepared from bovine blood to a final purity of approximately 90% IgG. Blood was also processed to produce a fraction containing elevated concentrations of IGF-I and TGF-beta (GF). Both IGC and GF were spray dried and blended with other ingredients to produce CR (30% IgG) or CS (15% IgG) containing 0 or 5% GF. Holstein bull calves were assigned to one of five treatments: 1) Pooled colostrum (MC)--1.9 L of pooled colostrum at 1 and 8 h of age to provide over 150 g of IgG; 2) Low supplement (LC)--1.9 L of CS at 1 and 8 h of age to provide 150 g of IgG; 3) Low supplement + GF (LG)--1.9 L of a CS with GF at 1 and 8 h of age to provide 150 g of IgG; 4) High supplement (HC)--1.9 L of CR at 1 h of age to provide 150 g of IgG and 1.9 L of a commercial milk replacer (MR) at 8 h of age; and 5) High supplement + GF (HG)--1.9 L of a CR with GF at 1 h of age to provide 150 g of IgG and 1.9 L of MR at 8 h of age. Apparent efficiency of IgG absorption was higher for calves fed HC and HG compared with those fed LC and LG and was lower for calves fed LG and HG compared with those fed LC and HC. IgG concentrations at 24 h were highest in calves fed MC compared with other calves and were higher in calves fed HC and HG compared to LC and LG. Calves fed LG and HG had lower IgG concentrations at 24 h of age compared with those fed LC and HC. These results indicate that 150 g of IgG provided in one dose soon after birth is superior to 150 g of IgG fed in two doses 7 h apart. Also, addition of a plasma fraction containing elevated concentrations of IGF-I and TGF-beta decreased IgG absorption in the neonatal calf. PMID- 14765817 TI - Methane production by mixed ruminal cultures incubated in dual-flow fermentors. AB - This study evaluated the effects of dilution rate and forage-to-concentrate ratio on gas production by rumen microbes. Continuous cultures were used to monitor methane production at three liquid dilution rates (3.2, 6.3, or 12.5%/h) and three forage-to-concentrate ratios (70:30, 50:50, or 30:70). Filtered ruminal contents were allowed 6 d of adaptation to diets followed by 7 d of data collection. Forage consisted of pelleted alfalfa and the concentrate mix included ground corn, soybean meal, and a mineral and vitamin premix. The experiment was replicated in a split-plot design. Total volatile fatty acid production averaged 58.0 mmol/d and was not affected by treatment. Molar proportion of acetate increased with increasing forage-to-concentrate ratio. Molar proportion of propionate tended to decrease at dilution rate of 12.5%/h and increased with the medium and low forage-to-concentrate ratio. Culture pH tended to be greater at a dilution rate of 12.5%/h. Methane production that was calculated from stoichiometric equations was not affected by treatments. However, methane production based on methane concentration in fermentor headspace resulted in an interaction effect of treatments. Stoichiometric equations underestimated methane output at higher dilution rates and with high forage diets. Total diet fermentability was lowest at dilution rate of 3.2%/h. Increasing dilution rates increased microbial yield; increasing the proportion of concentrate improved microbial efficiency. Dilution rate and forage-to-concentrate ratio altered the partition of substrate by microbes. Methane production based on actual concentrations differed from values estimated using stoichiometry of end-product appearance. PMID- 14765818 TI - Effects of feeding ground or steam-flaked broom sorghum and ground barley on performance of dairy cows in midlactation. AB - Ten Holstein cows in midlactation were used in a 5 x 5 replicated Latin square design with 21-d periods to determine the effects of feeding ground or steam flaked broom sorghum (Sorghum bicolor) and ground barley (Hordeum vulgare L.) on lactation performance and nutrient digestibility. Diets were fed as total mixed ration and consisted of 46% forage and 54% concentrate (DM basis). Treatment diets included ground barley, ground barley plus ground broom sorghum, ground broom sorghum, ground barley plus steam-flaked broom sorghum, and steam-flaked broom sorghum. Yield of fat-corrected milk was 2.3 kg greater for cows fed diets containing steam-flaked broom sorghum than for cows fed its ground form (24.4 vs 22.1 kg) and was 2.8 kg greater for cows fed diets containing a blend of steam flaked broom sorghum plus ground barley than for cows fed ground sorghum (24.9 vs 22.1 kg). Yields and percentages of milk fat, protein, SNF, total solids, and apparent digestibility of crude protein were greater for cows fed steam-flaked broom sorghum and ground barley vs. ground broom sorghum. Including steam-flaked rather than ground broom sorghum in diets significantly increased fecal pH (7.10 vs 6.87) and improved efficiency of feed conversion (1.26 vs 1.15). Feeding steam flaked broom sorghum alone or with ground barley compared with ground sorghum or the blend of ground barley and ground broom sorghum decreased plasma urea nitrogen increased glucose in plasma. Results of this study showed that feeding steam-flaked broom sorghum compared with ground broom sorghum could supply a more efficient source of energy for lactating cows. PMID- 14765819 TI - Comparison of conventional linted cottonseed and mechanically delinted cottonseed in diets for dairy cows. AB - Performance of lactating dairy cows fed diets containing either mechanically delinted whole cottonseed (DWCS; 3.7% lint) or linted whole cottonseed (LWCS; 11.7% lint) was measured. Forty primiparous (86 +/- 39 d in milk) and 40 multiparous (88 +/- 30 d in milk) cows were fed a total mixed ration containing 13% (dry matter basis) DWCS or LWCS in two blocks of 112 d (n = 53 and 27, respectively). Other total mixed ration ingredients (dry matter basis) were corn silage (28.1%), alfalfa silage (23%), high moisture shelled corn (27.8%), soybean meal (1.8%), expeller soybean meal (1.8%), blood meal (2%), and mineral-vitamin supplements (2.5%). Dry matter intake and milk yield were measured daily and milk composition every other week. Fecal grab samples were taken during wk 3 and 13 of each block to estimate excretion of intact whole cottonseeds. Milk yield, 3.5% fat-corrected milk, energy-corrected milk, milk composition and dry matter intake were not affected by whole cottonseed source. Body condition score tended to increase more with DWCS (0.22 vs. 0.11) for primiparous cows, although this was not reflected in body weight change. Dry matter digestibilities, based on indigestible ADF, were 63.5 and 64.8% for the DWCS and LWCS diets. It was calculated that 2.5 and 1.5% of the consumed seeds were excreted as whole cottonseeds in feces with the DWCS and LWCS diets, respectively. Although statistically significant, treatment differences in the proportion of intact seeds in the fecal DM would have little nutritional consequence. Mechanically delinted WCS performed as well as LWCS for all of the cow performance and milk composition variables measured. PMID- 14765820 TI - Effect of dietary phosphorus on performance of lactating dairy cows: milk production and cow health. AB - The objective of this study was to measure cow response to feeding of two dietary concentrations of P, one of which was close to recent National Research Council requirements, and the other of which was well in excess of the requirement. Diets containing 0.37 or 0.57% P (dry basis) were fed to Holstein cows for the first 165 d of lactation, and occasionally longer until cows were confirmed pregnant approximately 60 d after insemination. At calving, cows were randomly assigned to experimental diets. The number of cows completing a minimum of 165 d of lactation was 123 for the 0.37 and 124 for the 0.57% P groups. Cows were housed in a stanchion barn and fed one of two transition diets, each formulated to contain one of the P treatments for the first 3 wk of lactation, and then cows were moved to a free-stall barn where the experimental diets were group fed. Milk production, milk fat, and milk protein averaged 35.1 kg/d, 3.92%, and 2.90% for the 0.37% P diet, and 34.9 kg/d, 3.98%, and 2.91% for the 0.57% P diet. None of these measures were different between treatments. Blood serum P concentrations on d 50 and 100 of lactation averaged 6.1 and 6.2 mg/dL for the 0.37% P diet, and 6.8 and 6.9 mg/dL for the 0.57% P diet. No treatment differences were detected in milk production, cow health, or body condition score. PMID- 14765821 TI - Reproductive performance of dairy cows fed two concentrations of phosphorus. AB - The objective of this study was to determine the effect of dietary P concentrations of 0.37 (recommended) or 0.57% (excess; dry matter basis) on reproductive performance. At calving, Holstein cows were randomly assigned to 1 of 2 dietary treatments (n = 134 for 0.37% P and n = 133 for 0.57% P). Cows were fitted with a radiotelemetric transmitter (50 d in milk [DIM]) to record mounting activity during estrus and bred to natural estrus from 50 to 100 DIM, then to synchronized estrus (Ovsynch protocol) after 100 DIM. Weekly ultrasonography was performed from 50 DIM until pregnancy was diagnosed (-30 d after artificial insemination). Pregnancy was confirmed approximately 60 d after artificial insemination (artificial insemination). Weekly blood samples were analyzed for progesterone concentrations. Days to first increase (>1 ng/ml) in progesterone, days to first estrus detected by radiotelemetry, days to first service detected by herd personnel, and conception rate at first service did not differ between the recommended and excess P groups, respectively. Similarly, conception rate at 30 d, days open, pregnancies lost from 30 to 60 d, multiple ovulation rate, and the incidence of anovulatory condition at 71 DIM did not differ between these groups. The mean duration of estrus was 8.7 +/- 0.5 and 8.7 +/- 0.7 h, and the average number of mounts per estrus was 7.4 +/- 0.5 and 7.8 +/- 0.5 for a total mounting time during estrus of 25.8 +/- 1.8 and 24.5 +/- 1.6 s for cows fed the recommended and excess P diet, respectively. Phosphorus treatment had no detectable effect on reproductive performance. PMID- 14765822 TI - Production and metabolic responses of periparturient Holstein cows to dietary conjugated linoleic acid and trans-octadecenoic acids. AB - Thirty-eight multiparous Holstein cows were utilized in a completely randomized design to examine the effect of feeding calcium salts of conjugated linoleic acid (CLA) and trans-octadecenoic acids (trans-C18:1) on animal performance and lipid and glucose metabolism during the transition to lactation. Dietary treatments were initiated approximately 28 d prior to expected calving dates and continued through d 49 postpartum. Prepartum treatments consisted of 1) a basal diet (Control), 2) basal diet + 150 g/d of CLA mix (CLA), and 3) basal diet + 150 g/d of trans-C18:1 mix (TRANS). Amounts of calcium salts of CLA and trans-C18:1 mixes were adjusted to 225 g/d during the 49-d postpartum treatment period. All diets were offered as a total mixed ration. Prepartum fat supplementation had no detectable effects on dry matter intake, body weight, or body condition score. After parturition, cows in the TRANS group consumed less dry matter at wk 4, 5, and 6 of lactation than did cows in the control group. Cows fed the trans-C18:1 supplement were in a more severe negative energy balance than those fed the control diet at 1 wk of lactation. Periparturient fat supplementation had no detectable effects on milk yield during wk 1 to 7 of lactation. Milk fat was not affected during wk 1 to 4, but was reduced after wk 4 of lactation by dietary CLA. Feeding calcium salts of CLA decreased short- to medium-chain fatty acid (C4 to C14) concentrations and increased both linoleic and linolenic acid concentrations in milk fat. Concentrations of nonesterified fatty acids and beta hydroxybutyric acid in blood were greater in cows fed the CLA-supplemented diet than in those fed the control diet at 1 wk of lactation. In spite of small numerical tendencies, hepatic lipid and triacylglycerol concentrations did not vary significantly among dietary treatments. Periparturient fat supplementation had no detectable effects on plasma glucose and insulin concentrations. Steady state concentrations of hepatic mRNA encoding pyruvate carboxylase and phosphoenolpyruvate carboxykinase were greater for the TRANS treatment group than the control and CLA groups. Results indicate that dietary CLA and trans-C18:1 fatty acids may affect lipid and glucose metabolism in early postpartum Holstein cows through distinct mechanisms. PMID- 14765823 TI - Technical note: Estimating parameters of nonlinear segmented models. AB - The objective of this technical note is to develop an applied technique to estimate parameters using the Statistical Analysis System's nonlinear procedure (SAS PROC NLIN) for segmented models that have a change point or lag as one of the parameters. The goal is to select good starting values for the parameters near the global minimum or at least near a lower local minimum compared with what might be achieved using traditional starting values. The model used was f1 (t) = B1 + B4 if t < or = B3 and f2(t) = B1*exp[-B2*(t - B3)] + B4 if t > B3, where B1, B2, B3, and B4 are the parameters that require estimates, B3 is the change point or lag, and t = time. This technical note illustrates the solution when a traditional grid search for starting values is used and demonstrates a modified technique where starting values are systemically determined by fixing B3 over a range of reasonable values and then using the parameters from the solution with the lowest residual sums of squares as the starting values for the final solution. The modified method resulted in a lower square root of mean square error compared with the traditional method. The estimates for B3 (lag) were 3.5 for the modified method compared with 4.5 for the traditional method. This technique works well when using the SAS PROC NLIN procedure but can be modified to work with other statistical packages. PMID- 14765824 TI - Technical note: Comparison of chromatographic profile of glycomacropeptide from cheese whey isolated using different methods. AB - Glycomacropeptide (GMP) has heterogeneous carbohydrates, and this attributes to its various biological activities. In this study, we compared the chromatographic profiles of GMP isolated by three methods (trichloracetic acid fractionation, ethanol precipitation, and ultrafiltration) from whey protein isolate (WPI). Seven sharp heterogeneous GMP peaks were eluted from GMP prepared by ethanol precipitation and ultrafiltration using Mono Q anionic chromatography, while only 5 peaks were seen in TCA treated sample. The TCA pretreatment recovered only sialo-GMP (glycosylated) and eliminated all contaminated proteins; however, the recovery rate was the lowest (6.7% of the initial WPI). Ethanol precipitation recovered 20.4% of GMP from WPI and 75.7% was glycosylated, but the heating process might lead to degradation of glycosidic residues. Ultrafiltration was found to be the most effective in recovering GMP. The recovery rate was 33.9% with 81.6% sialo-GMP. We concluded that carbohydrate profile of GMP varied widely and depended on the isolation method. Based on the high recovery of sialo-GMP, the combination of ultrafiltration and anionic chromatography might be a suitable and practical approach on an industrial scale. PMID- 14765825 TI - Evaluation of serotypes of Staphylococcus aureus strains used in the production of a bovine mastitis bacterin. AB - The five Staphylococcus aureus strains used in the manufacture of a commercially available bacterin were examined for capsular and surface polysaccharide serotypes. Double immunodiffusion assays of antigenic extracts of test and reference strains with monospecific typing sera to capsular serotypes 1, 2, 5, and 8 and to surface polysaccharide serotype 336 were performed to detect the specific reactivities and antigenic relationships of test samples. Antigenic extracts of two S. aureus strains reacted with antibodies to serotype 8, but not with antibodies to serotype 5, by producing specific precipitin lines. A third strain reacted with monospecific antibodies to serotype 5 and not with the antibodies to serotype 8. The extracts of two other strains failed to exhibit any detectable reaction with antiserum to serotypes 1, 2, 5, or 8. Antibodies to serotype 336, however, precipitated an identical, specific 336 antigen from the antigenic extracts of these two nontypeable strains. Thus, S. aureus bacterin includes one serotype 5, two serotype 8, and two serotype 336 strains, the three predominant serotypes responsible for bovine mastitis. PMID- 14765826 TI - DairyBeef: maximizing quality and profits--a consistent food safety message. AB - To respond to meat safety and quality issues in dairy market cattle, a collaborative project team for 7 western states was established to develop educational resources providing a consistent meat safety and quality message to dairy producers, farm advisors, and veterinarians. The team produced an educational website and CD-ROM course that included videos, narrated slide sets, and on-farm tools. The objectives of this course were: 1) to help producers and their advisors understand market cattle food safety and quality issues, 2) help maintain markets for these cows, and 3) help producers identify ways to improve the quality of dairy cattle going to slaughter. DairyBeef. Maximizing Quality & Profits consists of 6 sections, including 4 core segments. Successful completion of quizzes following each core segment is required for participants to receive a certificate of completion. A formative evaluation of the program revealed the necessity for minor content and technological changes with the web-based course. All evaluators considered the materials relevant to dairy producers. After editing, course availability was enabled in February, 2003. Between February and May, 2003, 21 individuals received certificates of completion. PMID- 14765827 TI - Management practices and reported antimicrobial usage on conventional and organic dairy farms. AB - The primary objective was to compare reported antimicrobial usage between conventional and organic dairy farms. A secondary objective was to contrast selected management characteristics of conventional and organic dairy herds. A questionnaire was administered on site to selected dairy farmers located in Michigan, Minnesota, New York, and Wisconsin. Organic herds (n = 32) were smaller and produced less milk than conventional herds (n = 99). Lactating cows in organic dairies were more likely to be housed in tie stalls, whereas most conventional dairies housed cows in free stalls and milked in a parlor. Total mixed rations and purchased feeds were used on more conventional dairy farms compared with organic dairy farms. Conventional dairy producers were more likely to use advice from veterinarians for recommendations of treatment, and organic dairy producers were more likely to rely on advice from other farmers. Based on recall of antibiotic usage in the previous 60 d, 5.1, 84.9, 9.1, and 0.9% of farmers with conventional herds reported treatment of none, 1 to 10%, 11 to 25%, and >25% of milk cows, respectively. Most organic farmers (90.6%) reported no antibiotic treatments of milk cows, whereas 9.4% reported treating 1 to 10% of milk cows. Ceftiofur was the most commonly reported antibiotic for both farm types. Milk replacer containing antibiotics was reportedly used on 49.5% of conventional herds but only on one organic herd (3.1%). Antibiotics were used in heifer calves on 74.7% of conventional herds versus 21.9% of organic herds. Antibiotics to treat mastitis were used on 79.8% of conventional herds but on none of the organic herds. Most organic farms were in compliance with standards in advance of implementation of regulations. PMID- 14765828 TI - Pharmacokinetics of marbofloxacin in lactating cows after repeated intramuscular administrations and pharmacodynamics against mastitis isolated strains. AB - The plasma and milk pharmacokinetics of marbofloxacin, a fluoroquinolone antibacterial compound, were evaluated in dairy cows, as well as its pharmacodynamic characteristics against mastitis-isolated pathogens. Marbofloxacin was given intramuscularly as a 10% aqueous solution to dairy cows either at a single dose or at repeated doses of 2 mg/kg once daily for 3 d. Blood and milk samples were collected for the determination of the concentration of marbofloxacin and of its putative metabolites: N-desmethyl-marbofloxacin and N oxide-marbofloxacin. Bacterial field isolates were from milk samples collected from dairy cows suspected of having an intramammary infection. After identification, the minimal inhibitory concentration (MIC) was determined against the isolated strains. The maximal marbofloxacin concentration (Cmax) observed in milk after the first administration was 1.024 microg/mL, and the area under the curve during the first dosing interval was 6.513 microg/h per milliliter. After the third administration, these parameters were slightly increased (about 20% at most). Both metabolites were detected in the milk, but their concentrations were below the limit of quantification. The MIC against 90% of the population (MIC90) of Escherichia coli was 0.016 microg/mL, and it was 0.229 microg/mL against Staphylococcus aureus. The following surrogate clinical outcome markers were obtained against E. coli strains: a Cmax/MIC ratio of 67 and an area under the curve/MIC ratio of 407 h. Hence, a possible efficacy of marbofloxacin in the treatment of E. coli-induced mastitis could be expected as the endpoints of 10 and 250 h, respectively, are reached. PMID- 14765829 TI - Relationships of efficiency to reproductive disorders in Danish milk production: a stochastic frontier analysis. AB - Relationships of various reproductive disorders and milk production performance of Danish dairy farms were investigated. A stochastic frontier production function was estimated using data collected in 1998 from 514 Danish dairy farms. Measures of farm-level milk production efficiency relative to this production frontier were obtained, and relationships between milk production efficiency and the incidence risk of reproductive disorders were examined. There were moderate positive relationships between milk production efficiency and retained placenta, induction of estrus, uterine infections, ovarian cysts, and induction of birth. Inclusion of reproductive management variables showed that these moderate relationships disappeared, but directions of coefficients for almost all those variables remained the same. Dystocia showed a weak negative correlation with milk production efficiency. Farms that were mainly managed by young farmers had the highest average efficiency scores. The estimated milk losses due to inefficiency averaged 1142, 488, and 256 kg of energy-corrected milk per cow, respectively, for low-, medium-, and high-efficiency herds. It is concluded that the availability of younger cows, which enabled farmers to replace cows with reproductive disorders, contributed to high cow productivity in efficient farms. Thus, a high replacement rate more than compensates for the possible negative effect of reproductive disorders. The use of frontier production and efficiency/inefficiency functions to analyze herd data may enable dairy advisors to identify inefficient herds and to simulate the effect of alternative management procedures on the individual herd's efficiency. PMID- 14765830 TI - Effects of an automatic postmilking teat dipping system on new intramammary infections and iodine in milk. AB - A technology of automatically applying a postmilking teat dip via the milking machine prior to machine detachment was compared to manual postmilking teat dipping with a teat dip cup for effects on new IMI and iodine content in milk. One hundred twenty Holstein cows were experimentally challenged in a 22-wk trial with Streptococcus agalactiae and Staphylococcus aureus and 148 Holstein cows were experimentally challenged with Streptococcus uberis in another 22-wk trial. The bacterial suspensions were applied to teats of all of the cows after premilking udder preparation and immediately prior to milking machine attachment. In both trials, cows were divided among four treatments: no postmilking teat dipping; manual postmilking teat dipping with a proven efficacious iodophor teat dip; manual postmilking teat dipping with an iodophor teat dip formulated for an automatic postmilking teat dipping system; and automatically postmilking teat dipping via milking machines with an iodophor teat dip formulated for the automatic postmilking teat dipping system. The postmilking teat dipping treatments reduced new Staph. aureus IMI by 64.5, 76.5, and 88.2%; new Strep. agalactiae IMI by 61.5, 77.8, and 94.4%; and new Strep. uberis IMI by 63.5, 82.5, and 93.8%, respectively, against the treatment of no postmilking teat dipping. The treatment applying the postmilking teat dip automatically via milking machines had the lowest number of new IMI caused by the three pathogens. Teat end and teat skin condition were characterized as normal at the end of the study with no differences between treatments. There were no differences with regard to iodine content in milk between treatments. PMID- 14765831 TI - Changing trends in valve surgery in Europe: 1991-2000. AB - BACKGROUND AND AIM OF THE STUDY: This study sought to elaborate the changing trends in valve surgery in Europe during the 1990s. METHODS: The databases of different national surgical societies, registries and governments and international organizations were consulted and the data obtained were analyzed. RESULTS: The population of Europe (excluding Russia, CIS and Turkey) increased by 2.29%, from 509.67 million in 1991 to 519.15 million in 2000. During this period, the volume of cardiac surgery increased in Europe by 101% to 413,520 operations (797 per 10(6) population). The volume of valve surgery increased by 63% to 85,076 (164 per 10(6) population). The largest volumes of valve surgery were performed in Germany, France, UK, Italy and Spain. The increase in valve surgery volume was maximum in the Baltic states (+287%) and least in Scandinavia (+30%). Valve surgery volume per center per year changed from 139 in 1991 to 140 in 2000. The increase in overall valve surgery volume was sustained by an increase in subsets of octogenarian patients, valve surgery combined with coronary artery surgery, and increased productivity of the emerging economies of countries such as the Czech Republic, Hungary and Poland. Valve surgery as a proportion of cardiac surgery in Europe changed from 25.95% in 1991 to 21% in 2000. In 2000, valve surgery with combined procedures constituted 6.7% of total cardiac surgical volume, but 32% of valve surgical output in Europe. Mechanical valves have continued to dominate and were used in 77% of cases in 2000. Increased use of bioprostheses in the elderly subset among affluent economies was balanced by an increased use of mechanical prostheses in younger patients in emerging facilities in the East and South. The capital-intensive innovations (viz. robotic valve surgery, minimally invasive valve surgery, bioengineered valves) found niches only in some West European centers. Catheter-based procedures did not fulfil their promise. Balloon aortic valve dilatation investigations decreased drastically by the end of the decade, and balloon mitral dilatations number. Conservative aortic valve surgery is not yet practiced widely across Europe, while mitral repair has become widely accepted in clinical parlance. CONCLUSION: Despite the greater political and economic integration of Europe, the pattern of valve surgery continues to remain extremely diverse within the continent. PMID- 14765832 TI - Rheumatic heart disease occurrence, patterns and clinical correlates in children aged less than five years. AB - BACKGROUND AND AIM OF THE STUDY: Rheumatic fever (RF) and chronic rheumatic heart disease (RHD) are common in developing countries. Two-thirds of RHD patients are school-children aged between 5 and 15 years. Pre-schoolers aged <5 years are not immune to RF however, and to date RHD patterns in this very young age group have not been studied systematically. METHODS: Records of all RHD patients seen at the authors' institution between January 1999 and December 2000 were retrospectively reviewed. A special analysis was conducted among pre-school children aged <5 years. RESULTS: Thirty-eight (6.8%) of the RF/RHD admissions were aged <5 years, and 28 of these patients (20 males, 8 females) presented with acute RF. The mean age of acute RF diagnosis was 4 years. All RF/RHD patients aged <5 years were in normal sinus rhythm. Joint pain and swelling (25 cases; characteristic migratory polyarthritis in six, monoarthritis in five) and fever (24 cases) were the most frequent symptoms. Arthritis, carditis and chorea occurred in 75%, 50% and 4% respectively, with no instances of erythema marginatum or subcutaneous nodules. Effort intolerance, chest discomfort and palpitations were reported by nine, five and three cases, respectively. Mitral regurgitation was the most common valvular lesion in RF. The youngest case of confirmed acute RF was an 18-month-old male. The only patient with mitral stenosis in the present series was a 4-year-old girl. None of the patients required surgical intervention, and there were no deaths. CONCLUSION: RHD is common in very young age groups of <5 years. Pre schoolers account for a significant proportion of acute RF and chronic RHD admissions among children. Mitral regurgitation is the most common cardiac manifestation, but obstructive valve disease is distinctly rare in this age group. Aortic regurgitation, left ventricular dysfunction and pulmonary hypertension may complicate the course of RF in these very young children. PMID- 14765833 TI - Racial and ethnic differences in warfarin response. AB - BACKGROUND AND AIM OF THE STUDY: Variability of drug response among individuals is a well-recognized problem that may result in either under- or overtreatment of patients receiving similar drug concentrations. Patients with mechanical heart valves are dependent on adequate anticoagulation to prevent thrombosis development. 'Crystalline warfarin sodium' (warfarin) is the most common antithrombotic drug prescribed to control blood hemostasis in those patients, and also in those with indications such as stroke, myocardial infarction, pulmonary embolism and atrial fibrillation. Warfarin is a narrow therapeutic index agent; a small change in systemic concentration of the drug may lead to significant changes in pharmacodynamic response. Careful clinical management is required to balance the risks of bleeding (over-anticoagulation) with those of thrombosis (under-anticoagulation). The study aim was to summarize environmental, genetic and ethnic factors that affect a patient's response to warfarin and which must be considered for optimal patient outcome. METHODS: A Medline search was carried out to summarize various factors that influence a patient's response to warfarin. RESULTS: Inter-ethnic differences may have profound implications for the efficacy and safety of warfarin. Ethnic differences can affect pharmacokinetic features such as bioavailability, protein binding and volume of distribution, as well as hepatic metabolism and renal elimination. Environmental factors and genetic variants in human enzymes that metabolize warfarin also contribute to interindividual variations and may render some patients more susceptible to serious or life-threatening adverse events. CONCLUSION: Warfarin use is complicated by an unpredictable dose response that depends on factors such as demographics, diet, interacting drugs, genetic polymorphism and ethnic differences. The impact of racial differences on the kinetics of dose response or on drug efficacy is not well defined, as few clinical trials take ethnic variation into account. The use of the point of care and frequent patient self testing may permit standardized warfarin monitoring across diverse geographical regions and facilitate analysis of ethnic variation among subpopulations. PMID- 14765834 TI - Warfarin and herbal products interaction causing prosthetic aortic valve thrombosis presenting as acute myocardial infarction. AB - Myocardial infarction (MI) due to coronary artery embolization is a rare and potentially lethal complication of prosthetic heart valve thrombosis. A 58-year old man in whom the aortic valve was replaced with a bileaflet mechanical valve presented with an acute anterior MI. Valvular dysfunction was detected by physical examination, and confirmed by two-dimensional echocardiography and cinefluoroscopy. Coronary angiography disclosed embolization of the left anterior descending artery. Thrombotic encroachment of one of the prosthetic valve leaflets was found at reoperation. Failure to achieve adequate anticoagulation was likely due to an interaction between warfarin and herbal products. These findings have significant implications regarding the diagnosis and treatment of acute MI in patients with left-sided prosthetic heart valves, and emphasizes the importance of appropriate anticoagulation in this setting. PMID- 14765835 TI - Warfarin and cranberry juice: an interaction? AB - Herein, the case is reported of a persistently elevated INR consequent to an interaction between warfarin and cranberry juice in a patient with a prosthetic mitral valve. The elevation in INR was noted two weeks after the patient began to drink cranberry juice. Subsequent symptoms included postoperative bleeding problems. The bleeding was thought due to a biologically plausible interaction between the drug and the juice. It was concluded that patients taking warfarin should limit their cranberry juice intake. PMID- 14765836 TI - Left atrial volumes, function and work before and after mitral valve repair in chronic mitral regurgitation. AB - BACKGROUND AND AIM OF THE STUDY: Despite the effect of mitral valve repair in left ventricular (LV) function having been extensively studied, investigations of left atrial (LA) performance indices are minimal. This prospective study was undertaken to analyze LA volumes, function and work in patients with chronic mitral valve regurgitation (MR) who underwent mitral valve repair; the analyses were conducted both before and six months after surgery. METHODS: Twenty patients (15 males, five females; mean age 51.4 +/- 12.5 years) with severe MR (grade IV) due to floppy mitral valve/mitral valve prolapse (FMV/MVP; anterior, posterior or both) underwent mitral valve repair. LA volumes, maximal at mitral valve opening (LAmax); minimal at valve closure (LAmin); and at onset of atrial systole (P-wave on ECG, LAP); and transmitral Doppler A-wave velocity were measured before and six months after surgery. LA stroke volume (LASV) = LAP - LAmin; LA ejection fraction (LAEF) = LASV/LAP; LA kinetic energy (LAKE) = 1/2 x LASV x 1.06 (specific gravity of blood) x A2 (dyne x cm x 10(3)); LA and LV dimensions and functions were assessed at the same time. RESULTS: NYHA functional class was improved postoperatively by at least one grade. LV systolic and diastolic dimensions were reduced significantly in all patients (p <0.001). LA volumes (LAmax, LAmin and LAP) were decreased significantly in all patients (p <0.001); LASV remained unchanged. LAEF and LAKE were increased significantly (both p <0.001). The A-wave was also increased (p <0.001). CONCLUSION: Increased LA work (LAKE) after mitral valve repair, despite a decrease in LA volumes, suggests that LA muscle dysfunction was present before surgery. LA involvement may precede LV involvement. The determination of LA performance and work will help to optimize the timing of surgery in patients with FMV/MVP and MVR. PMID- 14765837 TI - The renin-angiotensin system genetic polymorphisms and rheumatic mitral valve disease. AB - BACKGROUND AND AIM OF THE STUDY: Angiotensin-converting enzyme (ACE) gene insertion/deletion (I/D) polymorphism, angiotensinogen (AGT) gene polymorphism and angiotensin II type 1 receptor (AT1R) polymorphism in relation to rheumatic mitral valve disease were examined in a case-control study to investigate possible relationships between these gene polymorphisms and rheumatic mitral valve disease in patients undergoing mitral valve replacement (MVR). METHODS: A total of 50 patients with rheumatic mitral valve disease and undergoing MVR was compared with 50 normal, and age- and sex-matched control subjects. ACE I/D, AGT gene M235T and AT1R-adenine/cytosine 1166 (A1166C) genotype polymorphisms were identified by polymerase chain reaction (PCR) -based restriction analysis. RESULTS: ACE I/D polymorphism differed significantly between the groups. The control group mostly represented the heterozygote ID allele (74%), while the MVR group showed frequencies of 60% for the homozygote DD and II alleles. MM homozygote frequency was significantly greater in controls, but TT homozygote frequency was significantly greater in the MVR group. AT1R-A1166C genotype polymorphism also differed significantly between groups; the MVR group had 73.7% of the AC heterozygote allele, while controls had 64.4% of the AA and 66.7% of the CC homozygote alleles. CONCLUSION: These results provided evidence of an association between ACE I/D polymorphism, M235T polymorphism and AT1R-A1166C genotype polymorphism and rheumatic mitral valve disease. PMID- 14765838 TI - Comparison of echocardiographic methods in assessing severity of mitral regurgitation in patients with mitral valve prolapse. AB - BACKGROUND AND AIM OF THE STUDY: Mitral regurgitation (MR) shows different characteristics in mitral valve prolapse (MVP); hence, it is important to assess MR severity accurately in these patients. The study aim was to compare Doppler echocardiographic methods in making such assessment. METHODS: Forty-seven patients with confirmed MVP and at least moderate mitral insufficiency, as established by Doppler echocardiography, were studied. Quantitative Doppler was used as the reference standard method. Color Doppler mapping was used to determine regurgitant jet area (JA/LAA), flow convergence (EROA-PISA) and vena contracta width (VCW). Systolic pulmonary venous flow reversal (SPVFR) and mitral E-wave velocity were also monitored. RESULTS: Univariate analysis showed severe MR to be significantly correlated to age, presence of atrial fibrillation, left ventricular systolic and diastolic diameter, left atrial diameter, mitral E velocity, JA/LAA, VCW, EROA-PISA and the presence of SPVFR. On multivariate analysis, the strongest determinants of severe MR were EROA-PISA, VCW and E velocity. The greatest area under the receiver-operator curve for diagnosing severe MR was observed with EROA-PISA. The 45-mm2 threshold of EROA-PISA had the highest risk ratio of severe MR with a high sum of sensitivity and specificity. However, the JA/LAA had the lowest risk ratio and negative predictive value for severe MR. CONCLUSION: PISA, VCW, E velocity and SPVFR measurements may be used to evaluate MR severity semi-quantitatively in patients with MVP; however, the ratio of JA/LAA appears to be a less reliable method in this respect. PMID- 14765839 TI - Doppler echocardiographic evaluation of right ventricular diastolic function in patients with mitral regurgitation. AB - BACKGROUND AND AIM OF THE STUDY: The effect of left ventricular (LV) pressure overload on right ventricular (RV) diastolic function has been extensively studied. In contrast, no data are available concerning the influence of LV volume overload on RV diastolic function. Accordingly, RV diastolic function was studied in patients with mitral regurgitation (MR) using Doppler echocardiography. METHODS: RV diastolic indices were calculated, using pulsed Doppler echocardiography, in 30 patients (mean age 56.87 +/- 8.58 years) with severe MR, and in 30 healthy control subjects (mean age 56.67 +/- 8.52 years). RESULTS: Compared with controls, MR patients had a significantly lower RV E/A ratio (0.85 +/- 0.12 versus 1.21 +/- 0.16, p <0.001), a significantly prolonged RV isovolumic relaxation time (70 +/- 20 versus 30 +/- 10 ms, p <0.001), a significantly prolonged deceleration time of the transtricuspid E wave (210 +/- 20 versus 140 +/- 10 ms, p <0.001), and a significantly greater right atrial filling fraction (38.58 +/- 4.59 versus 32.58 +/- 3.14%, p <0.001). There was no statistically significant correlation between RV diastolic indices and LV mass index and interventricular septum thickness. CONCLUSION: RV diastolic function in patients with MR is impaired, reflecting prolonged relaxation and redistribution of RV filling into late diastole. Ventricular interdependence constitutes the most likely mechanism of this action. PMID- 14765840 TI - Comparison of immediate and mid-term results of mitral balloon valvotomy in children and adolescents with those in adults. AB - BACKGROUND AND AIM OF THE STUDY: The study aim was to evaluate and compare the hemodynamic benefits of mitral balloon valvotomy (MBV) in young (child/adolescent) and older (adult) patients with severe mitral stenosis. METHODS: The younger group comprised 61 patients (mean age 17 +/- 3 years), and the older group 264 patients (mean age 35 +/- 10 years). All patients underwent MBV using an Inoue balloon to treat symptomatic and severe mitral stenosis. The long-term follow up of both groups was monitored on a comparative basis. RESULTS: The success rate of MBV was similar in both groups (95% in younger patients, 96% in older patients) (p = 0.36). Mitral valve area (MVA) was increased from 0.8 +/- 0.2 to 2.0 +/- 0.3 cm2 in the younger group, and from 0.8 +/- 0.3 to 1.9 +/- 0.3 cm2 in the older group. The mean mitral valve gradient was decreased significantly both in the younger group (from 13 +/- 5 to 5 +/- 2 mmHg) and older group (from 12 +/- 5 to 5 +/- 3 mmHg) (p < 0.001). Both groups had similar MVA and mean gradient at baseline and follow up. At follow up (mean 37 +/- 27 months), MVA monitored by Doppler echocardiography remained at 1.7 +/- 0.4 and 1.6 +/- 0.4 cm2 (p = 0.08). Similarly, the mitral valve mean gradient remained stable (p = 0.92). Severe mitral regurgitation developed in one younger patient, and in seven older patients (p = 0.66). Two cases of tamponade occurred in each group, and one older patient suffered a transient stroke. There was no mortality in either group. CONCLUSION: In patients with rheumatic mitral stenosis, the outcome of MBV in children and adolescents was at least comparable with that in adults. In addition, the benefits were similar in each age group on completion of follow up. PMID- 14765841 TI - Is there a role for the left ventricle apical-aortic conduit for acquired aortic stenosis? AB - BACKGROUND AND AIM OF THE STUDY: Aortic valve replacement (AVR) in patients with a heavily calcified ascending aorta and aortic root, or with conditions that preclude a median sternotomy, poses a formidable challenge. A left ventricle apical-aortic conduit (AAC) is an alternative in these situations. Herein, the authors' experience with AAC in adult patients with acquired aortic stenosis is reported. METHODS: Between 1995 and 2003, 13 patients (mean age 71 years) underwent AAC for severe symptomatic aortic stenosis (mean valve area 0.65 +/- 0.02 cm2). Indications for AAC were heavily calcified ascending aorta and aortic root (n = 5), patent retrosternal mammary grafts (n = 4), calcified ascending aorta and aortic root plus patent retrosternal mammary graft (n = 1), retrosternal colonic interposition (n = 1) and multiple previous sternotomies (n = 2). Seven patients had previous coronary artery bypass grafting (CABG). The mean preoperative left ventricular ejection fraction was 50 +/- 4%. RESULTS: AAC were performed under cardiopulmonary bypass through a left thoracotomy (n = 10), median sternotomy (n = 2) or bilateral thoracotomy (n = 1). Hearts were kept beating (n = 5) or fibrillated (n = 7). Circulatory arrest was used in one patient. Composite Dacron conduits with biological (n = 6), mechanical (n = 4) or homograft (n = 2) valves were used. Distal anastomoses were performed in the descending thoracic aorta (n = 12) or in the left iliac artery (n = 1). Two patients underwent simultaneous CABG. Three patients died in-hospital from ventricular failure (n = 1), intravascular thrombosis (n = 1) and multi-organ failure (n = 1). The mean hospital stay was 26 days. Complications included respiratory failure requiring tracheostomy (n = 2), stroke (n = 1) and re exploration for bleeding (n = 2). At a mean follow up of 2.1 years, there have been four late deaths; causes of death were congestive heart failure (n = 2), ischemic cardiomyopathy (n = 1) and cancer (n = 1). CONCLUSION: AAC provides an acceptable alternative to AVR in selected patients who are at exceedingly high risk for the standard procedure. PMID- 14765842 TI - Durability and prevalence of aortic regurgitation nine years after aortic valve replacement with the Toronto SPV stentless bioprosthesis. AB - BACKGROUND AND AIM OF THE STUDY: Stentless aortic bioprostheses have excellent hemodynamics. Previous investigations of the Toronto SPV valve described a correlation between the occurrence of significant aortic regurgitation (AR) and dilation of the sinotubular junction. The study aim was to determine the long term durability and determinants of AR at nine years in a large, multicenter study of the Toronto SPV valve. METHODS: The study included 447 patients from six centers. Clinical outcomes and echocardiographic data (gradients, effective orifice area index (EOA-I), left ventricular mass, aortic root dimensions, and presence and severity of AR) were collected prospectively. A multivariable logistic regression model was used to evaluate clinical and echocardiographic variables for impact on the occurrence of AR. RESULTS: Total follow up was 2,660 patient-years (mean 6.0 +/- 2.5 years; range: 0 to 11.1 years). Mean gradient and EOA-I remained unchanged through nine years. There were 17 cases of structural deterioration, of which 15 underwent explantation. The mechanism of failure was predominantly leaflet tear in the setting of sinotubular dilation. Freedom from explant for structural failure was 90.1% at nine years (100% for patients aged > or = 65 years). Freedom from hemodynamically significant AR was 96.9% at five years and 82.5% at nine years. Determinants of AR were longer duration of follow up, larger valve size, and increase in the ratio of sinotubular junction to the size of valve implanted. CONCLUSION: At nine years after implantation of the Toronto SPV valve, hemodynamics remained excellent. There was good freedom from structural deterioration through nine years, and structural failure occurred due to aortic root dilation and leaflet tear, without significant valve calcification. AR tends to occur with longer follow up, larger valve sizes, and dilation of the sinotubular junction. PMID- 14765843 TI - The Ross procedure as the surgical treatment of active aortic valve endocarditis. AB - BACKGROUND AND AIM OF THE STUDY: The authors' experience is reported of aortic valve replacement (AVR) using the pulmonary autograft in patients with active aortic valve endocarditis, including an urgent Ross procedure in infants with the acute condition. METHODS: Nine patients aged between 8 months and 38 years, with a diagnosis of aortic valve endocarditis, have undergone AVR using the Ross procedure at the authors' institution since October 1997. The diagnosis was established by clinical and echocardiographic findings. Indications for surgery were severe aortic insufficiency and congestive heart failure in all patients, with the addition of thromboembolic events (n = 3), persistent hyperpyrexia (n = 3) and vegetations (n = 5). Four infants with no history of congenital cardiac malformation underwent urgent surgery because of acute bacterial endocarditis and rapid hemodynamic deterioration. Blood cultures were positive for Streptococcus pneumoniae in three patients, and Kingella kingi and Staphylococcus aureus in one patient each. Four patients were culture-negative. All patients were treated with intravenous antibiotics for four to six weeks postoperatively. RESULTS: There were no perioperative or late deaths, and no recurrent endocarditis at the implanted valves. Echocardiographic evaluation at discharge showed trivial to mild aortic insufficiency, with no stenosis at the left ventricular outflow tract. Similar findings were found across the right ventricular outflow tract. At follow up (range: 4 months to 5.5 years), none of the patients showed progression of aortic valve insufficiency or developed stenosis; three had mild and moderate homograft stenosis (Doppler gradient 20-40 mmHg), and all children had moderate homograft insufficiency. CONCLUSION: The Ross procedure is an excellent therapeutic option for active aortic valve endocarditis in young patients, and demonstrates low morbidity and mortality. Early surgery may be indicated in patients with acute aortic valve endocarditis because of the rapidly progressive nature of this disease. PMID- 14765844 TI - Quantification of pulmonary autograft characteristics using magnetic resonance imaging. AB - BACKGROUND AND AIM OF THE STUDY: The diameters and distensibility of the native pulmonary root and their effect on pulmonary autograft performance were examined pre- and postoperatively using cardiac ultrasound and magnetic resonance imaging (MRI). METHODS: Eight patients undergoing the Ross procedure were prospectively involved. The diameters of the native aortic, native pulmonary and autograft roots were measured at the level of the annulus, sinus, sinotubular junction and in the main root using MRI through the cardiac cycle. Ultrasound was also used to estimate the degree of regurgitation, both pre- and postoperatively. RESULTS: The pulmonary root implanted into the systemic circulation increased in size but decreased in distensibility significantly at the sinus, sinotubular junction and main root, but not at the annulus. Postoperatively, the pulmonary autograft annulus showed a similar size and distensibility to that of the native aortic annulus. A wide range of aortic annular sizes (22-30 mm) produced clinically competent valves postoperatively. All undersized pulmonary valves showed only trivial regurgitation postoperatively. Although there was no clear correlation between root shape and valve insufficiency, two patients with mild and moderate autograft regurgitation both had divergent pulmonary roots (diameter at sinotubular junction > annulus diameter) preoperatively. CONCLUSION: The pulmonary autograft using the root replacement technique functioned well in all but one case. The shape of the native pulmonary root may be a determinant of early autograft regurgitation, as well as the diameter and the size mismatch between the great arteries. PMID- 14765845 TI - De Vega tricuspid annuloplasty for systemic tricuspid regurgitation in children with univentricular physiology. AB - BACKGROUND AND AIM OF THE STUDY: Significant tricuspid valve regurgitation (TR) is problematic in children with univentricular physiology and a systemic tricuspid valve occasionally requiring tricuspid (systemic atrioventricular) valve replacement. Since 1998, the De Vega tricuspid annuloplasty technique has been applied for TR in these children. METHODS: Twelve children (median age 2.2 years; range: 6 months to 17 years) with moderate or severe systemic TR underwent a De Vega tricuspid annuloplasty during a bidirectional Glenn anastomosis (n = 3), Fontan procedure (n = 8) or aortic valve replacement late after a Fontan procedure (n = 1). Nine patients (75%) had prior Norwood palliation for hypoplastic left heart syndrome. Four patients had simultaneous repair of an abnormal tricuspid valve in addition to the De Vega procedure. RESULTS: There were no deaths during a mean follow up of 2.0 +/- 1.4 years (range: 6 months to 5.1 years). One child required pacemaker implantation early after operation, and one child with a Glenn anastomosis underwent cardiac transplantation 21 months postoperatively. In the remaining 11 patients, the most recent echocardiogram showed mild or no TR in eight children, mild-to-moderate TR in one child, and moderate TR in two children. No child had symptomatic TR (including the two with moderate TR), significant tricuspid stenosis, or late pacemaker implantation. CONCLUSION: The De Vega tricuspid annuloplasty safely provides excellent relief of systemic TR in children with univentricular physiology, with a majority of patients (73%) having mild or less residual TR at follow up examination. This simple technique is preferred to tricuspid (systemic) valve replacement in these children. PMID- 14765846 TI - Forty-year survival with the Starr-Edwards heart valve prosthesis. AB - BACKGROUND AND AIM OF THE STUDY: The study aim was to update an analysis of the long-term survival of heart valve replacement using the Starr-Edwards prosthesis. METHODS: Cases of isolated aortic (AVR, n = 2,247) and mitral (MVR, n = 1,406) valve replacement with Starr-Edwards prostheses implanted between 1960 and 1997, with follow up to 2003, were reviewed. Introduced in 1965, the Models A1200/1260, M6120 are still in use (Current), while other models have been discontinued (Discontinued). For AVR, 938 valves were Discontinued, with a total follow up of 8,506 patient-years (pt-yr) and a maximum of 41 years; by comparison, 1,309 valves were Current, with a total follow up of 11,586 pt-yr and a maximum of 36.1 MVR, were Discontinued, with a total follow up of 6,454 pt-yr and maximum of 37.2 years; and 771 valves were Current, with a total follow up of 6,211 pt-yr and maximum of 37.0 years. RESULTS: Kaplan-Meier (KM) survival at 10 years was 53% for AVR and 51% for MVR; KM survival at 20 years was 23% for both AVR and MVR; KM survival at 30 years was 8% for both AVR and MVR; KM survival at 40 years was 4% for AVR. The standard error for all KM percentages was 1%. Four patients are currently alive with their original valves, more than 40 years after implantation. CONCLUSION: This series of patients, who underwent valve replacement with the Starr-Edwards valve, now have a follow up extending beyond 40 years, thereby confirming the excellent durability of this valve. PMID- 14765847 TI - A single-center experience of the Sorin Bicarbon heart valve prosthesis: long term clinical, hematological and hemodynamic results. AB - BACKGROUND AND AIM OF THE STUDY: The Sorin Bicarbon (SB) bileaflet valve, available since 1990, has a good clinical safety profile. The study aim was to assess the long-term clinical results following implantation, plus hemodynamic and hematological aspects of the valve. METHODS: A total of 195 SB prostheses (100 aortic, 67 mitral, 14 double valve replacements) was implanted in 181 patients (108 males, 73 females) since 1991. RESULTS: Mean age at implantation was 60.5 +/- 10.8 years. Actuarial analysis at 9.0 years showed an overall survival of 63.9 +/- 5.5%; 28.2% of deaths were early (n = 11), and 71.8% late (n = 28). Freedom from thrombosis was 99.4 +/- 0.6% (one early event, successfully reoperated on), and from embolic events was 86.3 +/- 4.7%. Freedom from endocarditis and hemorrhagic complications were respectively 98.1 +/- 1.1% and 98.6 +/- 0.9%. No occurrence of hemolysis was reported. Freedom from non structural valve dysfunction was 98.7 +/- 0.9%, and from reoperation was 96.1 +/- 1.6%. Hemodynamic evaluation by echocardiography and hematological testing was performed at 7.0 +/- 1.3 years (range 5-9 years) in a subset of 31 patients (20 males, 11 females). Mean valve size was 23.9 mm for aortic mitral. Echocardiography showed that aortic valves had a mean effective orifice area (EOA) of 2.26 +/- 1.28 cm2, with peak and mean pressure gradients of 23.6 +/- 14.6 and 12.9 +/- 8.1 mmHg, respectively. The mean EOA of mitral valve prostheses was 2.67 +/- 0.66 cm2, with peak and mean pressure gradients of 12.2 +/- 4.5 and 3.8 +/- 1.6 mmHg, respectively. Overall mean hemoglobin was 13.4 +/- 1.35 g/dl, and serum lactate dehydrogenase 630 +/- 13 IU/l. Serum haptoglobin was detectable in one patient only, and the mean reticulocyte count was 1.24 +/- 0.51%. CONCLUSION: In the present authors' experience, the Sorin Bicarbon prosthesis has low complication rates and maintains an excellent hemodynamic function over time. Hemolytic potential is insignificant with this valve. PMID- 14765848 TI - Mechanical valve prosthesis is a valid option for aortic valve replacement in the elderly. AB - BACKGROUND AND AIM OF THE STUDY: Thromboembolism and hemorrhage related to anticoagulation remain a major concern in elderly patients with mechanical valves. Clinical results following isolated aortic valve replacement (AVR) with tilting disk and bileaflet prostheses in patients aged over 70 years were analyzed and compared with results in patients aged <45 years. METHODS: Between January 1980 and August 2002, 319 consecutive older patients (group A) and 497 young patients AVR. Preoperative clinical data, early and late mortality, valve related complications and data related to anticoagulation status (including mean INR and mean interval between INR assays) were compared between groups. RESULTS: Hospital mortality was lower in group B (3.4%) than in group A (10.7%; p <0.0001). Twelve-year actuarial survival was lower in older patients (54% in group A versus 78% in group B; p <0.001). The two groups showed similar 12-year actuarial freedom from hemorrhage (99.6% versus 99.5%; p = 0.69), endocarditis (99.6% versus 98.43%; p = 0.25) and perivalvular leak (99.6% versus 97.9%; p = 0.21). However, actuarial freedom from thromboembolism was lower in older patients (98.8% versus 99.7%; p = 0.041). CONCLUSION: Despite lower rates of long term mortality and thromboembolism (the latter because of advanced atherosclerosis) in group A, there were no differences in rates of other valve related complications. Hence, older age cannot be considered a contraindication to implantation of mechanical valves in the aortic position. PMID- 14765850 TI - New laboratory technique measures projected dynamic area of prosthetic heart valves. AB - BACKGROUND AND AIM OF THE STUDY: Fluid dynamic forces, valve design factors and gravity interactively determine the complex motion of prosthetic heart valve occluders. Although motion has been investigated, in vitro, using high-speed image recording, the technique has significant cost and limitations on resolution. METHODS: The kinematics of mechanical and biological valve occluders in the aortic and mitral positions were assessed by measuring projected dynamic valve area (PDVA). Valves were tested in a pulse duplicator system simulating normal cardiac conditions. To quantify PDVA, light passage through back illuminated valves was measured by a calibrated photosensor with high-frequency response up to 150 kHz. Ten consecutive cycles were sampled using a PC data acquisition system. The system was calibrated under static conditions using reference areas. RESULTS: Several characteristics can be obtained from PDVA measurement including: maximum and minimum PDVA; rate of change of valve opening and closing PDVA; occluder rebound; and oscillatory open occluder behavior. Biological valves open more rapidly, close more gently, and exhibit no occluder rebound. They are also unaffected by gravity, and vary little in behavior from cycle to cycle compared with mechanical valves. CONCLUSION: A new method for measuring PDVA has been developed. Distinct differences in performance between valves were identified. It is hypothesized that, aside from patient factors and differences in materials, mechanical valves that mimic the PDVA behavior of biological valves, will lead to reduction of thrombogenicity, cavitation and high intensity transient signals (HITS), and also reduce sound level and regurgitation. PMID- 14765849 TI - Predictors of outcome in patients with prosthetic valve dysfunction. AB - BACKGROUND AND AIM OF THE STUDY: The study aim, based on the authors' experience in patients with prosthetic valve dysfunction, was to investigate risk factors for mortality and morbidity by analyzing preoperative, intraoperative and postoperative variables with respect to early and long-term survival. METHODS: A retrospective analysis was carried out of 132 patients (47 men, 85 women; mean age 46.8 +/- 12.4 years) who presented for treatment of prosthetic valve dysfunction between December 1992 and February 2003. Two patients received thrombolytic therapy and were excluded from the statistical analysis, which comprised only operatively treated patients; four patients underwent successful surgical repair of mitral mechanical prostheses; all other patients (except two who died perioperatively) underwent prosthetic valve re-replacement (n = 124). RESULTS: Overall mortality and hospital mortality rates were 15.2% and 10.6%, respectively. Postoperatively, 54 complications were seen in 42 patients (32.3%). Preoperative left ventricular end-systolic diameter (LVESD) > or = 45 mm and cardiopulmonary bypass (CPB) time > 140 min were independent risk factors for overall and in-hospital mortality. Female gender, age > 60 years and prolonged CPB time were predictors of postoperative complications. The actuarial survival rate was 87.5 +/- 0.3% at five years, and 81.7 +/- 0.4% at 10 years. A reduced left ventricular ejection fraction (LVEF) was the only independent predictor of late death and long-term survival. CONCLUSION: Preoperative LVESD > or = 45 mm and lower LVEF were found to be independent predictors of postoperative mortality and late survival, respectively. It is possible to obtain a substantial improvement in outcome and long-term survival if a valvular reoperation can be performed with shorter CPB time and before left ventricular dysfunction has developed. PMID- 14765851 TI - Valvular calcification in hemodialysis patients randomized to calcium-based phosphorus binders or sevelamer. AB - BACKGROUND AND AIM OF THE STUDY: Valvular calcification is common in patients with end-stage renal disease, and is associated with an unfavorable prognosis. It was hypothesized that sevelamer, a non-calcium-based phosphorus binder, might attenuate the progression of valvular calcification. METHODS: Two hundred subjects on maintenance hemodialysis received either sevelamer or calcium-based phosphorus binders. To assess the extent of calcification, 186 subjects underwent baseline electron beam tomography (EBT) of the coronary arteries, aorta and mitral and aortic valves, and 132 had follow up EBT scans at week 52. Changes in valvular calcification and combined valvular/vascular calcification were monitored and compared. RESULTS: At baseline, mitral valve calcification was seen in 46% of subjects, aortic valve calcification in 33%. Most subjects with zero values at baseline failed to progress over one year. Aortic valve calcification was significantly increased in calcium-treated subjects. Changes in mitral valve calcification, and combined mitral + aortic valve calcification were less in sevelamer-treated than in calcium-treated subjects, but not significantly so. When combining valvular and vascular calcification, the median (10%, 90%) change in sevelamer-treated subjects was significantly lower than in calcium-treated subjects (6, -5084 to 1180 versus 81, -1150 to 2944, p = 0.04). The effect of sevelamer remained significant after adjustment for baseline calcification and the time-averaged calcium-phosphorus product, and was independent of the calcium preparation (acetate versus carbonate), geographic region (US versus Europe), LDL or HDL-cholesterol, C-reactive protein and statin use. Significantly more sevelamer-treated subjects experienced an arrest (45 versus 28%, p = 0.047) or regression (26 versus 10%, p = 0.02) in total valvular and vascular calcification. CONCLUSION: Sevelamer arrested the progression of valvular and vascular calcification in almost 50% of hemodialysis subjects. Sevelamer treatment, plus intensive control of calcium and phosphorus levels, may attenuate progression of, or achieve regression in, cardiac valvular calcification. PMID- 14765852 TI - Surgical treatment of commissural prolapse: case report and review of the options. AB - Repair of commissural prolapse associated with chordal rupture may represent a challenging problem for the cardiac surgeon. The case of a patient with chronic mitral regurgitation and prolapse of the posterior commissural area associated with chordal rupture is presented. A technique was proposed that involved partial resection of the degenerative leaflet segment, plication of the posterior annulus and chordal transfer to create a neocommissure. An analysis of the options available for surgical treatment of commissural prolapse is included. PMID- 14765853 TI - Left ventricular outflow tract pseudoaneurysm after aortic valve replacement: case report. AB - A 68-year-old woman was admitted for angina pectoris and general fatigue without symptoms or signs of infective endocarditis. The patient had undergone re replacement of an aortic prosthetic valve three months previously. Transesophageal echocardiography revealed an echo-free cavity in the mitral aortic intervalvular fibrosa region just below the aortic annulus, communication of the echo-free cavity with the left ventricular outflow tract, and turbulent flow into the cavity. Left ventriculography revealed a cavity that arose just below the aortic prosthetic valve, and which expanded in systole and collapsed in diastole. Coronary angiography showed significant stenosis of the proximal right coronary artery, but neither stenoses nor compression were found in the left coronary artery. Patch closure of the pseudoaneurysm and aortic root replacement using a Freestyle valve with reconstruction of the coronary arteries were successfully performed. Surgical trauma to the intervalvular fibrosa during removal of the original prosthetic valve may have caused pseudoaneurysm formation in this patient. PMID- 14765854 TI - Mechanical heart valve prosthesis in the pulmonary position without anticoagulation: case report. AB - A 10-year-old girl underwent tetralogy of Fallot (TOF) repair and subsequent pulmonary valve replacement with a St. Jude Medical mechanical heart valve prosthesis. Valve replacement was necessary due to right heart failure resulting from pulmonary regurgitation occurring three months after TOF repair. At the age of 25 years, when she became pregnant, routine cardiac evaluation indicated that she had not used oral anticoagulation during the past 15 years. The patient was of rural origin, and of poor socioeconomic status, but is currently in her 15th postoperative year, with neither clinical problems nor any sign of valve failure. PMID- 14765855 TI - Aortic valve papillary fibroelastoma. AB - Papillary fibroelastomas are benign and rare tumors usually involving the cardiac valves. The diagnosis of this condition has gradually increased over the past few years. This is attributed to the increasing number of echocardiographic and transesophageal procedures that are currently performed. The case is presented of a woman diagnosed with an aortic valve mass, the sessile nature of which in the aortic root was considered a potential source of cerebral and coronary embolism. The mass was surgically removed, with no major complications being encountered perioperatively. PMID- 14765856 TI - Digital photography and computerisation in orthopaedics. PMID- 14765857 TI - Giant-cell tumour of bone. PMID- 14765858 TI - The management of periprosthetic fractures around the femoral stem. PMID- 14765859 TI - Changes in bone mineral density in the proximal femur after cementless total hip arthroplasty. A five-year longitudinal study. AB - We investigated prospectively the bone mineral density (BMD) of the proximal femur after implantation of a tapered rectangular cementless stem in 100 patients with a mean age of 60 years (16 to 87). It was determined using dual energy x-ray absorptiometry, performed one week after surgery and then every six months until the end-point of five years. The BMD increased significantly in Gruen zones 2, 4 and 5 by 11%, 3% and 11% respectively, and decreased significantly in Gruen zones 1, 6 and 7 by 3%, 6% and 14% respectively, over the five-year period. The net mean BMD did not change over this time period. The changes in the BMD were not confined to the first 12 months after surgery. This investigation revealed no change in the overall periprosthetic BMD, but demonstrated a regional redistribution of bone mass from the proximal to distal zones. PMID- 14765860 TI - Pain in the well-fixed, aseptic titanium hip replacement. The role of corrosion. AB - We have investigated nine patients with cemented Furlong (JRI, London, UK) titanium hip replacements who presented with early pain despite a well-fixed, aseptic prosthesis. All were followed up clinically and radiologically at regular intervals. Pain was located in the thigh and was worse at night. Radiographs showed cortical hypertrophy of the femur around the tip of the stem. Eight of the nine patients subsequently required single-stage revision using an uncemented prosthesis, which relieved the pain. At revision, the pH of the tip of the stem was found to be highly acidic with macroscopic evidence of corrosion consisting of multiple layers of titanium oxides when studied by X-ray dispersive analysis. Cemented titanium implants have a potential for crevice corrosion leading to cortical hypertrophy and intractable pain. PMID- 14765861 TI - Treatment of anaemia after joint replacement. A double-blind, randomised, controlled trial of ferrous sulphate versus placebo. AB - After total hip and knee replacement arthroplasty, patients may become anaemic and may be prescribed oral iron. There is, however, no published evidence that this is of benefit when used postoperatively. We treated 72 patients who were anaemic after primary total hip and knee arthroplasty by randomly allocating them to receive six weeks of either oral ferrous sulphate (35 patients) or a placebo (37 patients). Both groups of patients were similar in all aspects except for the treatment given. There was no statistically significant difference in the change of haemoglobin levels between the two groups. We therefore believe that the prescription of iron to all anaemic patients post-operatively should be avoided. The level of serum ferritin should be monitored at preoperative assessment. PMID- 14765862 TI - Premature wear and osteolysis in an HA-coated, uncemented total hip arthroplasty. AB - The ABG I cementless hip prosthesis has demonstrated unacceptably high rates of wear and osteolysis in our patients. We performed a retrospective study of 97 hips implanted between 1992 and 1998. Radiographic analysis revealed high rates of wear of the polyethylene liner with marked peri-acetabular osteolysis. Clinical examination indicated that many of these patients were initially asymptomatic. Wear-related problems have required ten hips to be revised and a furher 13 are awaiting revision. This gives a failure rate of 24% at a mean follow-up of 69 months. Contributing factors are likely to include poor wear characteristics of the polyethylene liners which were gamma irradiated in air, and increased wear debris caused by a poor fit of the polyethylene liner within the shell. We believe that all ABG I implants should be immediately reviewed and remain under careful, long-term follow-up. PMID- 14765863 TI - Arthroscopic debridement in the treatment of the infected total knee replacement. AB - Infection is a potentially disastrous complication of total knee replacement (TKR). Retention of the prosthesis has been associated with high rates of persistent infection. Our study shows that in selected situations, arthroscopic debridement may allow retention of the prosthesis and eradication of the infection. However, the prosthesis must be stable, the surgical technique must be meticulous and specific antibiotics must be taken for a lengthy period. Arthroscopic debridement should be considered as an alternative to an open technique, or revision, for the infected TKR. PMID- 14765864 TI - Intra-operative findings in varus osteoarthritis of the knee. An analysis of pre operative alignment in potential candidates for unicompartmental arthroplasty. AB - Interest in unicompartmental knee arthroplasty (UKA) for the treatment of medial compartment osteoarthritis has increased in recent years with apparent improvement in the long-term results. This is a result of improved surgical technique, patient selection, and implant design. In an effort further to improve patient selection we analysed the relationship between the pre-operative alignment of the knee and the anatomical findings at the time of surgery. We compared these findings with the indications for UKA. From 4021 total knee arthroplasties we compared intra-operative observations with the pre-operative clinical data in order to identify knees with isolated, medial, compartment changes, which would have been ideal candidates for UKA. We found that only 247 of the knees (6.1%) met anatomical qualifications for isolated, medial, unicompartmental osteoarthritis, and of these, only 168 (4.3%) met clinical standards ideal for UKA. Preoperative alignment showed a significant relationship with patterns of disease. Logistic regression revealed a relationship between pre operative alignment and intraoperative findings resembling a Gaussian distribution. Patients with a pre-operative varus alignment of 7 degrees were slightly more likely to be selected for UKA. But the further the anatomical alignment in either direction varies from 7 degrees of varus, the more unlikely it is for the knee to exhibit a disease pattern of isolated, medial, unicompartmental osteoarthritis. PMID- 14765865 TI - Treatment of recurrence of symptoms after excision of an interdigital neuroma. A retrospective review. AB - During a six-year period (January 1996 to January 2002), we re-explored 60 interspaces (49 patients, 49 feet) for recurrence or persistent symptoms after one or more previous procedures for excision of an interdigital neuroma. Ten patients underwent concomitant excision of a primary neuroma from an adjacent interspace, and 19 underwent concomitant forefoot surgery. The mean follow-up was 39.7 months (6 to 79). Evaluation included review of records and radiographs, clinical assessment, and a questionnaire regarding satisfaction, pain, restriction of footwear and activity. In total, 15 patients (30.7%) were completely satisfied, 13 (26.5%) were satisfied with minor reservations, ten (20.4%) were satisfied with major reservations and 11 (22.4%) were dissatisfied with the outcome. Of the 49 patients, 28 (57.2%) had no or mild pain, 29 (59.2%) had moderate or severe restriction of footwear and eight (16.3%) had moderate restriction of activity. Intra-operative findings, simultaneous surgery to adjacent interspaces, concomitant forefoot surgery and previous re-explorations did not significantly influence the outcome. Persistent or recurrent symptoms after transection of a nerve present a challenging problem for both the surgeon and patient. It is essential that there is a thorough pre-operative discussion with the patient, providing the rates of failure and the increased likelihood of restriction of footwear and activity after revision surgery. PMID- 14765866 TI - Sensory dysfunction in the great toe in hallux valgus. AB - Injury to the dorsomedial cutaneous nerve in the foot may occur after operations for hallux valgus. Pressure neuropathy before operation is also described but remains largely unexplored. We have investigated the incidence of sensory deficit in the great toe before operating for hallux valgus and examined to what extent any deficit was related to the degree of angulation of the joint. Forty-three patients with a total of 61 great toes with hallux valgus presenting for consideration of surgical correction had their sensation tested in pre-designated zones using a five-filament set of Semmes-Weinstein monofilaments. These allowed good inter-observer reliability with an intra-class correlation coefficient of 0.84. Sensory symptoms were noted by only 21% of the patients, a measurable reduction in sensation by one monofilament grade or more was found in an additional 44%. No relationship was found between the degree of sensory loss and the degree of angulation. Patients with symptomatic hallux valgus may have sensory loss in the toe without being aware of it. Normal subjective sensation does not reliably predict normal sensory function. Given the potentially high rates of nerve damage following operations for hallux valgus, we recommend objective sensory testing as part of routine assessment before surgery. PMID- 14765867 TI - The prognosis following acute primary glenohumeral dislocation. AB - We have studied 105 patients with 107 acute, primary, dislocations of the glenohumeral joint seen between January 1, 1991 and July 1, 1994. The mean time of follow-up was 71 months (46 to 91). In 34% the injury occurred during a sports activity and in 28% at home. The bias toward sport was even greater in patients less than 40 years of age, and in men. In patients older than 40 years of age, and in women, the dislocation occurred more often at home. The overall probability of recurrence within four years was 26%. Age was the most significant prognostic factor in recurrence which took place in 64% of patients less than 20 years of age and in 6% of those older than 40 years. Statistically, there was no difference between the rates of recurrence in patients who were active in sport and those who were not. The mean Rowe score for the whole group was 87 (15 to 100). Associated fractures were found in 20 patients (19%) and nerve injuries in 22 (21%). None of those in whom a fracture of the greater tuberosity was seen subsequently suffered a recurrent dislocation. At follow-up we found that 36 patients (34%) had not returned to their former employment but in only 2% was this owing to the injured shoulder. PMID- 14765868 TI - The management of infection in arthroplasty of the shoulder. AB - The management and outcome of treatment in 42 patients (49 shoulders) with an infected shoulder prosthesis was reviewed in a retrospective multicentre study of 2343 prostheses. The factors which were analysed included the primary diagnosis, the delay between the diagnosis of infection and treatment and the type of treatment. Treatment was considered to be successful in 30 patients (71%). Previous surgery and radiotherapy were identified as risk factors for the development of infection. All patients with an infected prosthesis had pain and limitation of movement and 88% showed radiological loosening. In 50% of the shoulders, the antibiotics chosen and the length of treatment were considered not to be optimal. The mean follow-up was 34 months. Antibiotics or debridement alone were ineffective. In acute infection, immediate revision with excision of all infected tissue and exchange of the prosthesis with appropriate antibiotic therapy gave the best results. Multidisciplinary collaboration is recommended. PMID- 14765869 TI - Choline acetyltransferase activity and evoked spinal cord potentials for diagnosis of brachial plexus injury. AB - The purpose of this study is to investigate the diagnostic value of evoked spinal cord potentials (ESCPs) and choline acetyltransferase (CAT) activity during exploration of injuries to the brachial plexus. We assessed 25 spinal roots in 19 patients. The results of the two investigations were consistent in all except two roots. Although assessment of ESCPs is easy and quick, it mainly records the nerve potentials along the sensory pathway. Although measurement of CAT activity needs a specimen of the nerve and the availability of a radioisotope laboratory, it gives direct information regarding the motor function of ventral spinal roots. These two techniques should be complementary to each other in order to achieve a more accurate diagnosis. PMID- 14765870 TI - Correlation of nerve root pain with dermatomal sensory threshold and back pain with spinal movement in single level lumbar spondylosis. AB - We studied 27 patients with low back pain and unilateral L5 or S1 spinal nerve root pain. Significant radiological changes were restricted to the symptomatic root level, when compared with controls. Low back and leg pain were graded on a visual analogue scale. Dermatomal quantitative sensory tests revealed significant elevations of warm, cool and touch perception thresholds in the affected dermatome, compared with controls. These elevations correlated with root pain (warm v L5 root pain; r = 0.88, p < 0.0001), but not with back pain. Low back pain correlated with restriction of anteroposterior spinal flexion (p = 0.02), but not with leg pain. A subset of 16 patients underwent decompressive surgery with improvement of pain scores, sensory thresholds and spinal mobility. A further 14 patients with back pain, multilevel nerve root symptoms and radiological changes were also studied. The only correlation found was of low back pain with spinal movement (p < 0.002). We conclude that, in patients with single level disease, dermatomal sensory threshold elevation and restriction of spinal movement are independent correlates of sciatica and low back pain. PMID- 14765871 TI - Functional recovery in patients with nonunion treated with the Ilizarov technique. AB - There are few reports on function after limb salvage surgery using the Ilizarov technique, and none that document the pattern of recovery or predict when maximum function returns. This prospective, longitudinal study documents the baseline functional abilities of 40 consecutive patients with nonunion of a fracture in the lower limb. Patients were studied for at least two and a half years following the completion of surgery. Function was measured by timed tests of functional performance and by the Toronto Extremity Salvage Score self-reported patient questionnaire. Recovery was slowest in the early stages after removal of the frame and greatest between six months and one year. Statistically significant improvement continued up to, but not beyond two years. This observation has important implications for the length of follow-up incorporated into the rehabilitation programmes for patients, predictions of patient status in regard to compensation and for the design of future studies to evaluate functional outcome. PMID- 14765872 TI - Treatment of unstable trochanteric fractures. Randomised comparison of the gamma nail and the proximal femoral nail. AB - The proximal femoral nail (PFN) is a recently introduced intramedullary system, designed to improve treatment of unstable trochanteric fractures of the hip. In a multicentre prospective clinical study, the intra-operative use, complications and outcome of treatment using the PFN (n = 211) were compared with those using the gamma nail (GN) (n = 213). The intra-operative blood loss was lower with the PFN (220 ml v 287 ml, p = 0.001). Post-operatively, more lateral protrusion of the hip screws of the PFN (7.6%) was documented, compared with the gamma nail (1.6%, p = 0.02). Most local complications were related to suboptimal reduction of the fracture and/or positioning of the implant. Functional outcome and consolidation were equal for both implants. Generally, the results of treatment of unstable trochanteric fractures were comparable for the PFN and GN. The pitfalls and complications were similar, and mainly surgeon- or fracture-related, rather than implant-related. PMID- 14765873 TI - Paradoxical cerebral embolisation. An explanation for fat embolism syndrome. AB - Fat embolism occurs following fractures of a long bone or arthroplasty. We investigated whether paradoxical embolisation through a venous-to-arterial circulation shunt (v-a) could lead to cerebral embolisation during elective hip or knee arthroplasty. Transcranial Doppler ultrasound (TCD), following the intravenous injection of microbubble contrast, identified the presence of a shunt in 41 patients undergoing hip (n = 20) or knee (n = 21) arthroplasty. Intra operative cerebral embolism was detected during continuous TCD monitoring. Of the 41 patients, 34 had a v-a shunt of whom 18 had an embolism and embolism only occurred in patients with a shunt (p = 0.012). Spontaneous and larger shunts were associated with a greater number of emboli (rs = 0.67 and rs = 0.71 respectively, p < 0.01). Observations in two patients with large spontaneous shunts revealed 368 and 203 emboli and unexplained post-operative confusion and pancreatitis. Paradoxical cerebral embolisation only occurred in patients with a shunt and may explain both postoperative confusion and fat embolism syndrome following surgery. PMID- 14765874 TI - Lower limb arthroplasty complicated by deep venous thrombosis. Prevalence and subjective outcome. AB - The aim of this study was to determine the prevalence of deep venous thrombosis (DVT) following lower limb arthroplasty and to assess whether this adversely affected satisfaction, relief from pain, or the level of mobility as perceived by patients. Six hundred and ten consecutive recipients of primary total hip replacement (THR) or total knee replacement (TKR) underwent routine post operative venography. The functional outcome had already been assessed at one year by using the Regional Arthroplasty Database questionnaire, the results of which were correlated to venographic records. The combined prevalence of DVT after THR and TKR in the patients, who did not receive chemical thromboprophylaxis, was 46.4%. Thrombus was identified in 57.6% of those with a TKR and in 33.5% of patients with a THR. Proximal thrombus was found in 11.0% of TKRs and in 14.8% of THRs. One year after surgery, patients who had a DVT established by venography did not report higher levels of immobility (p = 0.07), discomfort (p = 0.12) or dissatisfaction (p = 0.23) when compared to those with patent venous systems. This suggests that the prevalence of DVT following TKR/THR without chemical thromboprophylaxis is high and these findings are consistent with the literature. However, patients did not perceive thrombosis to compromise their overall outcome. This challenges the belief that DVT is associated with morbidity and calls for further comprehensive research in this area. The low morbidity of the lower limb associated with DVT in these patients does not support the use of chemical thromboprophylaxis. PMID- 14765875 TI - Conversion of biarticular to monoarticular muscles as a component of multilevel surgery in spastic diplegia. AB - We treated 20 children (40 limbs) with diplegic cerebral palsy who could walk by multilevel soft tissue operative procedures including conversion of the biarticular semitendinosus and gastrocnemius to monoarticular muscles. The mean age at surgery was 11.5 years (5.6 to 17.0). All patients underwent clinical and radiological examination and three-dimensional instrumented gait analysis before and at a mean of 3.1 years (2.0 to 4.5) after surgery. The passive range of movement at the ankle, knee and hip showed improvement at follow-up. Kinematic parameters indicated a reduced pelvic range of movement and improvement of extension of the knee in single stance after operation (p < 0.0001). However, post-operative back-kneeing was detected in five of the 40 limbs. The kinetic studies showed that the power of the hamstrings and plantar flexors of the ankle was maintained while the maximum knee extensor moment during stance was reduced. The elimination of knee flexor activity of semitendinosus and gastrocnemius combined with transfer of distal rectus femoris led to an improvement in gait as confirmed by gait analysis. PMID- 14765876 TI - Autologous transfusion in acetabuloplasty in children. AB - There were 22 patients with cerebral palsy aged six to 17 years who underwent an acetabuloplasty as part of an open reduction of the hip. In 11 patients a paediatric cell saver was used to collect autologous blood which was re-infused per-operatively. This group was compared to a cohort of 11 patients undergoing similar operations in whom only banked homologous blood was transfused. On average, 432 ml of autologous blood was reinfused compared to 909 ml of homologous blood (p < 0.01), representing 19.6% and 47% of the total blood volume, respectively (p < 0.002). Two units of homologous blood were transfused in the cell saver group compared with 20 units in the control group (p < 0.001). When using a paediatric cell saver, homologous blood transfusion was avoided in 82% of patients and there were no complications. PMID- 14765878 TI - Traumatic spondylolisthesis of the lumbosacral spine with multiple fractures of the posterior elements. AB - We describe a patient with a traumatic spondylolisthesis of L5 and multiple, bilateral pedicle fractures from L2 to L5. Conservative treatment was chosen, with eventual neurological recovery and bony union. We are not aware of previous reports of this pattern of injury. PMID- 14765877 TI - A malignant peripheral nerve-sheath tumour responding to chemotherapy. AB - A malignant peripheral nerve-sheath tumour developed in the right S1 nerve root in a man aged 30 causing back pain and sciatica. CT and MRI revealed a destructive tumour of the sacrum invading the retroperitoneal space. The tumour was not resectable with an adequate margin. Chemotherapy, consisting of high-dose ifosfamide followed by a combination of vincristine, doxorubicin and cyclophosphamide, was given with success. Malignant peripheral nerve-sheath tumours are thought to respond weakly to chemotherapy, but the response in our patient was complete. PMID- 14765879 TI - Biodegradation and biocompatability of a calcium sulphate-hydroxyapatite bone substitute. AB - An injectable material consisting of calcium sulphate mixed with hydroxyapatite was investigated as a possible alternative to autograft in the restoration of bone defects. The material was studied both in vitro in simulated body fluid (SBF) and in vivo when implanted in rat muscles and into the proximal tibiae of rabbits. Variation in the strength and weight of the material during ageing in SBF was measured. Tissue response, material resorption and bone ingrowth were studied in the animal models. A good tissue response was observed in both the rat muscles and rabbit tibiae without inflammatory reactions or the presence of fibrous tissue. Ageing in SBF showed that during the first week carbonated hydroxyapatite precipitated on the surfaces of the material and this may enhance bone ingrowth. PMID- 14765880 TI - Bone growth into a ceramic-filled defect around an implant. The response to transforming growth factor beta1. AB - Synthetic bone substitutes provide an alternative to autograft but do not give equivalent clinical results. Their performance may be enhanced by adding osteogenic growth factors. In this study, TGFbeta1 was absorbed on to a carrier of beta tricalcium phosphate and Gelfoam and used to fill a defect around a tibial implant in a rat model of revision arthoplasty. We added 0.0, 0.02 microg, 0.1 microg or 1.0 microg of TGFbeta1 to the carrier and then implanted it around an hydroxyapatite-coated stainless-steel pin in the proximal tibia of rats. The tibiae were harvested at three, six or 26 weeks and the amount of bone formation and ceramic resorption were assessed. TGFbeta1 had no effect on the amount of bone in the defect, the amount of fluorescent label incorporated or the rate of mineral apposition. The growth factor did not significantly affect the amount of betaTCP remaining in the tissue at any of the time points. PMID- 14765881 TI - Experimental posterolateral spinal fusion with porous ceramics and mesenchymal stem cells. AB - Alternatives to autogenous bone graft for spinal fusion have been investigated for many years. It has been shown that osteoconductive materials alone do not give a rate of fusion which is comparable to that of autogenous bone graft. We analysed the effectiveness of porous ceramic loaded with cultured mesenchymal stem cells as a new graft material for spinal fusion in an animal model. Posterolateral fusion was carried out at the L4/L5 level in 40 White New Zealand rabbits using one of the following graft materials: porous ceramic granules plus cultured mesenchymal stem cells (group I); ceramic granules plus fresh autogenous bone marrow (group II); ceramic granules alone (group II); and autogenous bone graft (group IV). The animals were killed eight weeks after surgery and the spines were evaluated radiographically, by a manual palpation test and by histological analysis. The rate of fusion was significantly higher in group I compared with group III and higher, but not significantly, in group I compared with groups II and IV. In group I histological analysis showed newly formed bone in contact with the implanted granules and highly cellular bone marrow between the newly formed trabecular bone. In group II, thin trabeculae of newly formed bone were present in the peripheral portion of the fusion mass. In group III, there was a reduced amount of newly formed bone and abundant fibrous tissue. In group IV, there were thin trabeculae of newly formed bone close to the decorticated transverse processes and dead trabecular bone in the central portion of the fusion mass. In vitro cultured mesenchymal stem cells may be loaded into porous ceramic to make a graft material for spinal fusion which appears to be more effective than porous ceramic alone. Further studies are needed to investigate the medium- to long-term results of this procedure, its feasibility in the clinical setting and the most appropriate carrier for mesenchymal stem cells. PMID- 14765882 TI - Concomitant tumour resistance in patients with osteosarcoma. A clue to a new therapeutic strategy. AB - Concomitant tumour resistance (CTR) is a unique phenomenon in which animals harbouring large primary tumours are resistant to the growth of smaller metastatic tumours by systemic angiogenic suppression. To examine this clinically, in ten patients with osteosarcoma, we investigated the effects of removal of the primary tumour on the development of pulmonary metastases, the systemic angiogenesis-inducing ability and the serum levels of several angiogenesis modulators. We found that removal of the primary tumour significantly elevated systemic angiogenesis-inducing ability in five patients who had post-operative recurrence of the tumour. Post-operative elevation of the angiogenesis-induced ability was suppressed by the addition of an angiogenic inhibitor, endostatin. Also, primary removal of the tumour decreased the serum levels of vascular endothelial growth factor and endostatin. These findings suggest, for the first time, the presence of CTR in patients with osteosarcoma for whom post-operative antiangiogenic therapy may be used to prevent the post operative progression of micrometastases. PMID- 14765883 TI - Catastrophic failure of the Elite Plus total hip replacement, with a Hylamer acetabulum and Zirconia ceramic femoral head. PMID- 14765884 TI - Internal fixation compared with total hip replacement for displaced femoral neck fractures in the elderly. PMID- 14765885 TI - Friction burns within the tibia during reaming. PMID- 14765886 TI - Turn-up bone flap for lengthening the below-knee amputation stump. PMID- 14765887 TI - Hyperbaric oxygen therapy as a treatment for stage-I avascular necrosis of the femoral head. PMID- 14765889 TI - Access to oral health care. PMID- 14765888 TI - Linear sebaceous naevus syndrome and resistant rickets. PMID- 14765890 TI - Periodontal disease and mortality in an aged population. AB - We investigated the relationship between periodontitis and mortality in home dwelling individuals aged (75+ years), considering potential confounders including C-reactive protein levels. This study involved 364 individuals, originally part of the Helsinki Ageing Study cohort, who underwent medical and dental examinations in 1990 and were followed for five years. After five years, 52 of the 175 dentate subjects had died. Using univariate analysis, the association between baseline periodontitis and mortality was of borderline significance. After controlling for the common risk factors, periodontitis more than doubled the risk of cardiovascular disease-related mortality (HR 2.28, CI 1.03-5.05). The increase in total mortality was, however, not statistically significant (HR 1.43, CI 0.81-2.50). Subjects who were edentulous had higher mortality than those who were dentate without periodontitis, but the difference was not statistically significant. A baseline CRP level above 3 mg/l was associated with high mortality; but this effect was significant only among individuals with periodontitis. These results suggest that periodontitis influences the pathogenesis and outcome of cardiovascular disease, especially in individuals who also have evidence of a systemic inflammatory reaction. PMID- 14765891 TI - Assessing prosthodontic dental treatment needs in older adults in Thailand: normative vs. sociodental approaches. AB - The authors compared the estimated prosthodontic treatment needs of edentulous older adults when using normative and sociodental approaches. This cross sectional study involved a non-random sample of 158 edentulous adults, aged 60 to 74 years, who were living independently in metropolitan Chiang Mai, Thailand. Each subject was given an oral examination and was interviewed. The dental questionnaire and the clinical criteria were adapted from the National Diet and Nutritional Survey for People Aged 65 and Older and included the Oral Impacts on Daily Performances sociodental indicator. Of the 158 subjects, we found using a normative assessment that 79.7% had a treatment need for complete dentures; 60.5% of those adults had impact-related treatment needs. Estimated treatment need dropped by about 40% when subjects were assessed using the sociodental approach. We concluded that large reductions in normative need estimates for complete dentures were apparent using a sociodental approach to assess prosthodontic dental treatment needs of older people. PMID- 14765892 TI - General anesthesia for developmentally disabled dental care patients: a comparison of reinforced laryngeal mask airway and endotracheal intubation anesthesia. AB - Providing dental care for developmentally disabled patients who require general anesthesia is challenging for both and dentists and anesthesiologists. This study aimed to compare the efficacy of two anesthetic methods for dental care. The researchers retrospectively analyzed morbidity data following anesthesia using either a reinforced laryngeal mask airway (LMA) or endotracheal intubation anesthesia for a two-year time period. The subjects were developmentally disabled patients receiving dental care. Statistical analyses were by unpaired student t tests and chi-square tests. Patients were who anesthetized with a reinforced laryngeal mask airway had a significantly shorter recovery period and lower postanesthetic complication rates when compared to patients undergoing endotracheal intubation anesthesia. Although hypoxemia (SPO2 < 90%) during dental care occurred more frequently when using the reinforced laryngeal mask airway, the difference was not significant. Nausea and vomiting were the major complications in the postanesthetic care unit and after discharge. When complication rates were compared in the two patient groups, nausea and vomiting were significantly higher during postanesthetic care and after discharge in the intubated group. In conclusion, reinforced laryngeal mask airway provides general anesthesia with less risk of side effects for developmentally disabled patients undergoing dental care. PMID- 14765893 TI - Prevalence and predictors of untreated caries and oral pain among Special Olympic athletes. AB - The authors assessed the prevalence and predictors of untreated caries and oral pain among Special Olympic athletes. The study population consisted of a convenience sample of 9,620 athletes who were participating in the 2001 Special Olympics events held at 40 sites in the United States, and who consented to a standardized oral health screening. The prevalence of oral pain and untreated caries was 13.5% and 30.4%, respectively. Statistically significant independent associations (adjusted odds ratios [OR], p < or = 0.05) between untreated caries and oral pain (OR = 1.50), gingivitis (OR = 1.92), injury (OR = 1.28), missing teeth (OR = 1.79), and home care (frequency of cleaning their teeth once or less per week compared with once or more per day OR = 2.13) were found. In another model, we found statistically significant independent associations between oral pain and untreated caries (OR = 1.58), gender (OR = 1.28), gingivitis (OR = 1.30), and home care (frequency of cleaning their teeth once or less per week compared to once or more per day OR = 4.60). Substantial levels of untreated caries and oral pain were prevalent and related to poor oral hygiene and poor oral health. These findings were discouraging given that the study participants represented a generally well-supported, high-functioning stratum of persons with mental retardation in the United States. PMID- 14765894 TI - Oral health needs and access to dental care of homeless adults in the United States: a review. AB - The homeless population in the United States is one of great diversity that continues to increase in number. Although data on the oral health status of individuals who are homeless is limited, studies consistently report both the perception and clinical evidence of dental needs among this population as well as a low utilization rate for dental services. This article reviews the oral health needs of people who are homeless as reported in literature, barriers to receiving dental care, and methods used to deliver dental care to this population. Many rehabilitation centers for adults who are homeless consider the establishment and maintenance of a state of good general and oral health as a priority and a key factor in helping homeless adults to return to the workforce and mainstream society. PMID- 14765895 TI - Prevalence of enteropathic Escherichia coli in dogs with acute and chronic diarrhoea. AB - Samples of faeces from 57 dogs with acute diarrhoea, 82 dogs with chronic diarrhoea, 34 clinically healthy household dogs and 88 kennelled control dogs were analysed by hybridisation, using DNA probes to detect enteropathogenic Escherichia coli (EPEC) and enterotoxigenic E coli (ETEC), verocytotoxin producing E coli (VTEC), enterohaemorrhagic E coli (EHEC), enteroinvasive E coli (EIEC) and enteroaggregative E coli (EAggEC). Samples of duodenal juice from 60 of the 82 dogs with chronic diarrhoea were also examined. Significantly more of the dogs with diarrhoea were excreting EPEC (acute 35.1 per cent, chronic 31.7 per cent) and VTEC (acute 24.6 per cent, chronic 28 per cent) than the kennelled dogs (EPEC 17.1 per cent, VTEC 0 per cent) or the household control dogs (EPEC 6 per cent, VTEC 5.9 per cent). Enteropathic E coli was also detected in the duodenal juice of 23 of 60 (38.3 per cent) of the dogs with chronic diarrhoea. The EPEC attaching and effacing A (eaeA) gene and the verocytotoxin 1 (VR1) gene coding for VTEC were often found together. There was good agreement between in vitro studies and hybridisation for the detection of eaeA and VT1. Isolates from the dogs with diarrhoea adhered significantly more to Hep-2 cells, and VT1 positive strains from the dogs with diarrhoea consistently killed more than 50 per cent of Vero cells. PMID- 14765896 TI - Treatment of suckling beef cattle with a progestagen sponge and oestradiol benzoate or equine chorionic gonadotrophin. AB - The ovarian responses of anoestrus beef cows to a combined treatment with medroxy progesterone acetate (MAP) sponges and oestradiol benzoate or equine chorionic gonadotrophin (eCG) were evaluated. Forty-five suckling Hereford cows were allocated to three equal groups. Group 1 received a MAP sponge for seven days plus an injection of 2 mg oestradiol benzoate when the sponge was inserted (day 0) and 1 mg when the sponge was withdrawn; group 2 received identical treatment until day 7, when a dose of 400 iu of eCG was administered, and group 3 were left untreated as control animals. From day 0 to day 11 the cows' ovaries were examined daily by transrectal ultrasonography, and their oestrous behaviour was observed from 24 hours to 96 hours after the sponge was removed. Data from cows that had a corpus luteum present before the sponge was withdrawn were not used in subsequent analyses; there were four in group 1, five in group 2 and four in group 3. In 19 of the 21 cows in groups 1 and 2 a new follicular wave was observed to emerge at a mean (sd) interval of 3.9 (0.3) days after the insertion of the sponge, whereas in group 3 it occurred in all 11 cows after 3.4 (0.6) days. Only the six cows that had a follicle of 9 mm or larger in diameter ovulated (P < or = 0.001). Nine of the 11 cows in group 1 came into oestrus, compared with two of the 10 in group 2 and none of the control cows (P < or = 0.001). Ovulation was observed in four, two and none of the cows in groups 1, 2 and 3, respectively. PMID- 14765897 TI - Avipoxvirus infection in peregrine falcons (Falco peregrinus) from a reintroduction programme in Germany. AB - Poxvirus infections are common in domestic birds in Germany, but they are rare in birds of prey. Only species of falconidae imported from Arabian or Asian countries have so far tested positive for poxvirus, and, among these, only raptors kept for falconry. As part of a reintroduction programme in the northern county of Mecklenburg-Western Pomerania, which is adjacent to the Baltic Sea, 21 young peregrine falcons were released into the wild; six of them died and one was examined postmortem, its tissues being examined by light and electron microscopy. In addition, an ELISA for fowlpox, pigeonpox and canarypox was applied. No virus could be isolated and propagation in culture failed, but virus particles were detected by electron microscopy in lesions from its skin and tongue. PMID- 14765898 TI - Preliminary evaluation of the prevalence of BSE in Japan. PMID- 14765899 TI - Isolation of a single genotype of bovine rotavirus in three geographically distant farms in Tunisia. PMID- 14765900 TI - Monitoring of canine dermatophytosis by an immunodiagnostic method. PMID- 14765901 TI - Onagers, a potential pasture parasite management tool in zoological collections. PMID- 14765903 TI - Veterinary surveillance. PMID- 14765902 TI - Isolation of Citrobacter murliniae from clinically ill and dead quail, ducks and chickens. PMID- 14765904 TI - Government veterinary surgeons website. PMID- 14765906 TI - 24-hour cover. PMID- 14765905 TI - Licensing requirements of Annex A species for CITES. PMID- 14765907 TI - Single intradermal tuberculin tests of milking buffaloes and cows in Nepal. PMID- 14765908 TI - Benefits of BVA membership. PMID- 14765909 TI - Improvement of chromatic aberration of a plastic rod lens array. 1. Combination color lens arrays consisting of several kinds of rod lenses with different gradient constants and different color filter functions. AB - We propose a new type of rod lens array with improved chromatic aberration. The new rod lens array consists of several kinds of rod lenses with different gradient constants and different color filter functions. The gradient constant of each lens was prepared such that the total conjugate length was identical to the specific wavelength of the lens with a constant lens length. We call our new type of rod lens array a combination-color lens array. The characteristics of this color-type lens array are as follows: high angular aperture, short focal length, and ease of production with established procedures. The optical resolution of the rod lens array is 300 dpi in a wide wavelength range that is high enough to be applied to color scanners. PMID- 14765910 TI - Analysis of capillary interferometry for measuring refractive indices of minute samples. AB - A method of measuring the refractive indices of minute samples by analyzing capillary interferometry is introduced. With the interference theory of light, the intensity distribution of an interference fringe pattern formed by a cylindrical tube of a capillary is obtained, and the influence of some parameters on the fringes are discussed. The measurement accuracy and its relative problems are analyzed. PMID- 14765911 TI - Double-grating interferometer for measurement of cylinder diameters. AB - We describe a double-grating interferometer for the measurement of cylinder diameters. The unique characteristic of this interferometer is that one can freely change the period of the interference fringes by turning the grating, which permits the measurement range of the interferometer also to be changed freely according to the cylinder diameter to be measured. A clear image of the cylinder can be obtained because the aperture diaphragm blocks the beams diffracted from the edge of the cylinder. The outside and inside diameters of the M4 x 0.7 mm hand tap are measured with this double-grating interferometer. PMID- 14765912 TI - Optical fiber probe for biomedical Raman spectroscopy. AB - In vitro experiments have demonstrated the ability of Raman spectroscopy to diagnose a wide variety of diseases. Recent in vivo investigations performed with optical fiber probes were promising but generally limited to easily accessible organs, often requiring relatively long collection times. We have implemented an optical design strategy to utilize system throughput fully by characterizing the Raman distribution from tissue. This scheme optimizes collection efficiency, minimizes noise, and has resulted in small-diameter, highly efficient Raman probes that are capable of collecting high-quality data in 1 s. Performance has been tested through simulations and experiments with tissue models and several in vitro tissue types, demonstrating that this new design can advance Raman spectroscopy as a clinically practical technique. PMID- 14765913 TI - Light propagation in biological tissues containing an absorbing plate. AB - We study light propagation in biological tissue containing an absorbing obstacle. In particular, we solve the infinite-domain problem in which an absorbing plate of negligible thickness prevents a portion of the light from the source from reaching the detector plane. Inasmuch as scattering in the medium is sharply peaked in the forward direction, we replace the governing radiative transport equation with the Fokker-Planck equation. The problem is solved first by application of the Kirchhoff approximation to determine the secondary source distribution over the surface of the plate. That result is propagated to the detector plane by use of Green's function. The Green's function is given as an expansion of plane-wave modes that are calculated numerically. The radiance is shown to obey Babinet's principle. Results from numerical computations that demonstrate this theory are shown. PMID- 14765914 TI - Phosphorescence lifetime imaging in turbid media: the inverse problem and experimental image reconstruction. AB - Three-dimensional phosphorescence lifetime imaging is a novel method for the mapping of oxygen concentration in biological tissues. We present reconstruction techniques for recovering phosphorescent objects in highly scattering media based on the telegraph equation and two regularization methods, i.e., the Tikhonov Phillips regularization and the maximum entropy method. Theoretical results are experimentally validated, and the reconstructed images of phosphorescent objects rendering oxygen maps in a layer are presented. PMID- 14765915 TI - Localized self-volume growth in hybrid sol-gel glass induced by ultraviolet radiation with a gray-scale mask. AB - UV-induced self-volume growth of the hybrid sol-gel glass was investigated for fabrication of an arbitrary-surface relief structure with a user-defined gray scale mask. The gray scale mask was generated through exposure of a high-energy beam-sensitive glass under an electron beam. Saw-tooth gratings of different periods were fabricated by a simple UV exposure through the gray-scale mask. Experimental results showed that the smaller the period is, the shallower the depth obtained owing to the self-volume growth effect is. PMID- 14765916 TI - Computer-aided lens assembly. AB - We propose a computer-aided method of lens manufacture that allows assembly, adjustment, and test phases to be run concurrently until an acceptable level of optical performance is reached. Misalignment of elements within a compound lens is determined by a comparison of the results of physical ray tracing by use of an array of Gaussian laser beams with numerically obtained geometric ray traces. An estimate of misalignment errors is made, and individual elements are adjusted in an iterative manner until performance criteria are achieved. The method is illustrated for the alignment of an air-spaced doublet. PMID- 14765917 TI - Polarized light scattering by dielectric and metallic spheres on oxidized silicon surfaces. AB - The polarization and intensity of light scattered by polystyrene latex and copper spheres with diameters of approximately 100 nm deposited onto silicon substrates containing various thicknesses of oxide films were measured with 532-nm light. The results are compared with a theory for scattering by a sphere on a surface, originally developed by others [Physica A 137, 209 (1986)] and extended to include coatings on the substrate. Nonlinear least-squares fits of the theory to the observations yield results that were consistent with differential mobility measurements of the particle diameter. PMID- 14765918 TI - Model of image degradation due to wind buffeting on an extremely large telescope. AB - A parametric model of wind buffeting on an extremely large telescope with a multipod secondary support is described. The model estimates wave-front errors that are due to wind buffeting on a segmented primary, wind-induced secondary figure and position errors, and primary-mirror deformations caused by wind forces on the secondary support. The approach is based on a Zernike expansion of pressure fluctuations, with simple models of stiffness, resonance, and control. The model shows that wind buffeting on a multipod attached to the primary mirror cell significantly degrades the image quality in a large telescope with a slow primary. PMID- 14765919 TI - Optical phased array configuration for an extremely large telescope. AB - Extremely large telescopes are currently under consideration by several groups in several countries. Extrapolation of current technology up to 30 m indicates a cost of over dollars 1 billion. Innovative concepts are being explored to find significant cost reductions. We explore the concept of an Optical Phased Array (OPA) telescope. Each element of the OPA is a separate Cassegrain telescope. Collimated beams from the array are sent via an associated set of delay lines to a central beam combiner. This array of small telescope elements offers the possibility of starting with a low-cost array of a few rings of elements, adding structure and additional Cass elements until the desired diameter telescope is attained. We address the salient features of such an extremely large telescope and cost elements relative to more conventional options. PMID- 14765920 TI - Measurement accuracy in control of segmented-mirror telescopes. AB - Design concepts for future large optical telescopes have highly segmented primary mirrors, with the out-of-plane degrees of freedom actively controlled. We estimate the contribution to errors in controlling the primary mirror that results from sensor noise and, in particular, compare mechanical measurements of relative segment motion with optical wave-front information. Data from the Keck telescopes are used to obtain realistic estimates of the achievable noise due to mechanical sensors. On the basis of these estimates, mechanical sensors will be more accurate than wave-front information for any of the telescope design concepts currently under consideration, and therefore supplemental wave-front sensors are not required for real-time figure control. Furthermore, control system errors due to sensor noise will not significantly degrade either seeing limited or diffraction-limited observations. PMID- 14765921 TI - Spectral matching factors between Super S25 and New S25 photo cathodes and reflective radiation of objects. AB - According to the response of photoelectric device to a light source, the formula of spectral matching factor of photo cathode-object combination is deduced. The spectral matching factors of Super S25 and New S25 photo cathodes for green vegetation and rough concrete are calculated and compared. For Super S251, the spectral-matching factors for green vegetation and rough concrete are 0.707 and 0.743, respectively, under the full-moon radiation and are 0.504 and 0.577, respectively, under the clear-star radiation. For Super S252, the full-moon factors are 0.561 (green vegetation) and 0.732 (rough concrete), and those for clear-star radiation are 0.338 (green vegetation) and 0.471 (rough concrete). For New S25, the full-moon factors are 0.741 (green vegetation) and 0.762 (rough concrete), and those for clear-star radiation are 0.603 (green vegetation) and 0.657 (rough concrete). An analysis of results shows that the spectral-matching factor has influence on the performance of a low-light level night vision system and is essential for evaluating the visual range of a night-vision system. PMID- 14765922 TI - Spot size, depth-of-focus, and diffraction ring intensity formulas for truncated Gaussian beams. AB - Simple polynomial formulas to calculate the FWHM and full width at 1/e2 intensity diffraction spot size and the depth of focus at a Strehl ratio of 0.8 and 0.5 as a function of a Gaussian beam truncation ratio and a system f-number are presented. Formulas are obtained by use of the numerical integration of a Huygens Fresnel diffraction integral and can be used to calculate the number of resolvable spots, the modulation transfer function, and the defocus tolerance of optical systems that employ laser beams. I also derived analytical formulas for the diffraction ring intensity as a function of the Gaussian beam truncation ratio and the system f-number. Such formulas can be used to estimate the diffraction-limited contrast of display and imaging systems. PMID- 14765923 TI - Effect of stress on performance of dense wavelength division multiplexing filters: optical properties. AB - Thin-film filters used for dense wavelength division multiplexing (DWDM) applications are processed by a variety of deposition techniques, including ion beam sputtering. Ion-beam sputtering produces high-quality coatings and provides flexibility of coating materials. However, DWDM filters consisting of oxide films that are reactively deposited by ion-beam sputtering, as in most sputter techniques, typically exhibit high levels of compressive stress. This affects the optical characteristics of the filters. Details of the filter passband characteristics and wave-front distortion illustrate the influence of the stress. Spatial variation of the stress on the filter surface causes the filter center wavelength to have spatial variation, and it causes the filter to have an asymmetric passband characteristic. PMID- 14765924 TI - Effect of stress on performance of dense wavelength division multiplexing filters: thermal properties. AB - Thin-film filters used for dense wavelength division multiplexing (DWDM) applications are processed by a variety of deposition techniques, including ion beam sputtering. Ion-beam sputtering produces high-quality coatings and provides flexibility of coating materials. However, DWDM filters consisting of oxide films that are reactively deposited by ion-beam sputtering, as in most sputter techniques, typically exhibit high levels of compressive stress. This affects the thermal characteristics of the filters. We have identified three thermal effects: center wavelength drift with temperature, center wavelength creep, and permanent center wavelength shift. The latter two are strongly dependent on the stress state of the filter. Models are presented that support the data that were taken. PMID- 14765925 TI - Semiactive infrared remote sensing: a practical prototype and field comparison. AB - A semiactive method of Fourier-transform infrared (FTIR) remote sensing has been developed and field tested. The method replaces the sender telescope of an active technique with an extended, heated broadband source. The output of the extended source (a commercial griddle) is not collimated and thus facilitates alignment by having the detector optics simply point at the griddle. The present source fills the detector's field of view at 100 m and maintains a temperature approximately 80 K warmer than ambient. In field tests with live CO releases, the method was approximately 30 times less sensitive than active methods, but approximately 30 times more sensitive than passive methods, with far greater sensitivity in the midwave infrared. PMID- 14765926 TI - Determination of the solar calibration constant for a sun-sky radiometer: proposal of an in-situ procedure. AB - A new procedure is presented for determining in situ the solar calibration constant, i.e., the Sun-sky radiometer counts for a direct normal solar flux extrapolated to the top of the atmosphere. The method makes use of a modified version of the Langley plot based on the use of an inversion code of column integrated aerosol size distribution, and it is ordinarily applied to calibrate Prede Sun-sky radiometers. To analyze how such an in situ method can work accurately, the technique has been applied to a five-month dataset obtained from measurements taken in Rome, Italy, by a Prede Sun-sky radiometer from 22 April to 5 November 2001. The precision of the in situ method has been estimated to within 1-2.5%, depending on the wavelength. PMID- 14765927 TI - Explicit formulas for transmission characteristics of graded-index oval core fibers. AB - Transmission characteristics of graded-index oval (GIO) core fibers are theoretically analyzed, and basic equations representing the specific properties of the fiber are obtained. Formulas for the propagation constant, the field profile function, the cutoff condition, and the total number of guided modes are derived in explicit forms. The formula for the change of the Gaussian beam radius as a function of the propagation distance is also given, and the numerical example agrees well with experimental data and agrees completely with that obtained by the beam-propagation method. These explicit expressions are useful for finding optimum structural parameters of GIO fibers to be used in practical fields. PMID- 14765928 TI - Effects of point-spread function on calibration and radiometric accuracy of CCD camera. AB - The point-spread function (PSF) of a camera can seriously affect the accuracy of radiometric calibration and measurement. We found that the PSF can produce a 3.7% difference between the apparent measured radiance of two plaques of different sizes with the same illumination. This difference can be removed by deconvolution with the measured PSF. To determine the PSF, many images of a collimated beam from a He-Ne laser are averaged. Since our optical system is focused at infinity, it should focus this source to a single pixel. Although the measured PSF is very sharp, dropping 4 and 6 orders of magnitude and 8 and 100 pixels away from the point source, respectively, we show that the effect of the PSF as far as 100 pixels away cannot be ignored without introducing an appreciable error to the calibration. We believe that the PSF should be taken into account in all optical systems to obtain accurate radiometric measurements. PMID- 14765929 TI - Design and fabrication of a glass waveguide optical add-drop multiplexer by use of an amorphous-silicon overlay distributed Bragg reflector. AB - We report the fabrication of a distributed Bragg reflector (DBR) on a silver ion exchanged glass waveguide with a patterned overlay of sputter-deposited amorphous silicon (a-Si). Using this high-refractive-index overlay DBR technology, we demonstrate a fully functional glass-integrated optic add-drop multiplexer (OADM). The OADM consists of a Mach-Zehnder interferometer (MZI) containing overlay DBR gratings in both arms. The design, fabrication, and characterization procedures of the sputter-deposited a-Si overlay DBR and the ion-exchanged glass waveguide MZI are discussed in detail. The completed 3-cm-long OADM exhibited a 24-dB transmission dip with a 3-dB bandwidth of 0.5 nm. The 1-cm-long a-Si overlay DBR induced an additional attenuation of 1.2 dB. A simplified method for overlay DBR fabrication that utilizes a lift-off technique is also proposed and demonstrated. PMID- 14765930 TI - Model of a multiple-line distributed-feedback dye laser. AB - The theoretical basis for simultaneous oscillation of 2N - 3 laser lines is due to interference of N (for all even N > or = 2) pump beams in a distributed feedback dye laser is described. Multiple gratings are produced in a dye solution by interference patterns of N/2 pairs of a frequency-doubled Nd:YAG laser. N/2 pairs of mutually time-delayed pulses induce multiple gratings of different periodicities, of which 2N - 3 gratings support oscillation of 2N - 3 lines and the remaining gratings, because of their larger periods, cannot support Bragg scattering. The maximum number of laser lines depends on the mutual delay between adjacent pairs of beams, coherence, states of polarization, pulse lengths, and of course the number of pulses. For three pairs of excitation beams derived from the same source through wave-front or amplitude phase division techniques, the output lasing lines varied from a minimum of three to a maximum of nine. This research was carried out by pumping of a dye solution with two, four, and six pulses, but the principle may be extended to multiple output lines, depending on the number of pump pulses and on the gain of the dye solution. PMID- 14765931 TI - Magnesium- and zinc-aluminosilicate cobalt-doped glass ceramics as saturable absorbers for diode-pumped 1.3-microm laser. AB - Q switching of a diode-pumped Nd3+:KGd(WO4)2 laser at 1.35 microm by use of cobalt-doped magnesium- and zinc-aluminosilicate glass ceramics as saturable absorbers is demonstrated. Q-switching efficiency up to 40% has been obtained. Ground-state absorption cross sections for Co2+ ions at the wavelengths of 1.35 and 1.54 microm are estimated to be (3.5-4.0) x 10(-19) cm2. Bleaching relaxation times under the excitation of the 4A2 --> 4T1(4F) transition of tetrahedrally coordinated Co2+ ions are of 280 +/- 50 ns and 700 +/- 80 ns for magnesium- and zinc-aluminosilicate glass ceramics, respectively. PMID- 14765932 TI - Production and characterization of spiral phase plates for optical wavelengths. AB - We describe the fabrication and characterization of a high-quality spiral phase plate as a device to generate optical vortices of low (3-5) specified charge at visible wavelengths. The manufacturing process is based on a molding technique and allows for the production of high-precision, smooth spiral phase plates as well as for their replication. An attractive feature of this process is that it permits the fabrication of nominally identical spiral phase plates made from different materials and thus yielding different vortex charges. When such a plate is inserted in the waist of a fundamental Gaussian beam, the resultant far-field intensity profile shows a rich vortex structure, in excellent agreement with diffraction calculations based on ideal spiral phase plates. Using a simple optical test, we show that the reproducibility of the manufacturing process is excellent. PMID- 14765933 TI - Wavelength calibration of spectra measured by the Global Ozone Monitoring Experiment: variations along orbits and in time. AB - The nadir-viewing Global Ozone Monitoring Experiment spectrometer aboard the second European Remote Sensing satellite measures spectra in the range of 240-790 nm. For the near-real-time delivery of ozone columns and profiles at the Royal Netherlands Meteorological Institute, a wavelength-calibration method was developed that allows for variation in both location and width of the spectral bins along the detector. The resultant wavelength grid of earthshine spectra varies along an orbit. This variation shows a correlation with instrument temperatures for a window near 306 nm; for other wavelength windows there is no correlation. The wavelength grid of calibrated solar spectra shows fluctuations without an apparent pattern and no correlation with instrument degradation. PMID- 14765934 TI - Neurological care in Pakistan: actions are needed. PMID- 14765935 TI - Future of neurology in Pakistan. PMID- 14765936 TI - Spectrum of complications and mortality of bacterial meningitis: an experience from a developing country. AB - OBJECTIVE: The aim of this study was to obtain data on predisposing factors, causative organisms and their associated mortality and complications related to acute bacterial meningitis. METHODS: The chart review of all patients in whom acute bacterial meningitis was diagnosed at The Aga Khan University Hospital from January 1995 through December 2001. RESULTS: One hundred ninety-four patients were included in study. There were 146 males and 48 females. The mean age of our study population was 41 +/- 12.3 years. One hundred and ninety (97.9%) patients had community-acquired meningitis; only 4 (2.0%) patients developed meningitis nosocomially. The two most common predisposing factors were diabetes mellitus (13.9%) and otitis media (7.7%) among all 194 patients. A significant proportion of patients with complications had diabetes mellitus (24.6%, p < 0.001). CSF and blood cultures were positive in 53 (27.3%) and 42 (21.6%) patients respectively; there was no statistical difference found. The most common organisms isolated were Streptococcus pneumoniae in 35 (36.8%) patients followed by Neisseria meningitides in 30 (31.5%) patients. Approximately 68% of positive cultures yielded S. pneumoniae and N. meningitides (p < 0.0001). The overall mortality rate was 22.1%. The mortality rate for Streptococcus pneumoniae was 17.1%. The highest mortality was observed in patients with Pseudomonal meningitis where all four patients expired followed by mortality rate of 85.7% in Escherichia coli afflicted patients (p < 0.001). Complications occurred in 73 (37.6%) patients with persistent complications in 31 (42.4%) patients. Complications resolved in 34 (46.5%) patients. The most common complications were seizures (12.8%) and cranial nerve palsies (11.3%). Seizures were more likely to occur in older patients (p < 0.05) whereas hydrocephalus was more common in younger patients (p < 0.05). CONCLUSION: Bacterial Meningitis remains a serious disease associated with substantial morbidity and mortality. Most cases are community acquired with S. Pneumoniae being the most common pathogen. Old age, diabetes mellitus, a positive culture, seizures as a complication and late stage in the disease are the important predictors of a poor outcome. PMID- 14765937 TI - Ischemic stroke subtypes in Pakistan: the Aga Khan University Stroke Data Bank. AB - OBJECTIVE: Frequency of ischemic stroke subtypes is influenced by ethnic and geographic variables. Our objective was to identify various stroke subtypes and its determinants at a tertiary care hospital. METHODS: We prospectively collected data on ischemic stroke subtypes admitted to The Aga Khan University Hospital in Karachi. RESULTS: A total of 596 patients were enrolled in 22 months in the Aga Khan Universtiy Stroke Registry. These included 393 patients with Ischemic stroke, 126 patients with intracerebral hemorrhage, 50 patients with subarachnoid hemorrhage and others. The ischemic stroke group was classified according to the TOAST criteria and comprised of lacunar 168/393 (42.7%); large artery atherosclerosis 106/393 (26.9%); cardioembolic 24/393 (6.1%); undetermined 80/393 (20.3%); and other determined types 15/393 (3.8%). The high proportion of lacunar strokes in our population may be due to high burden of inadequately treated hypertension and diabetes. Clear cut cardioembolic stroke was relatively infrequent in our population. CONCLUSION: Lacunar stroke is the most common subtype of stroke in our patient population. This is most likely secondary to uncontrolled hypertension. PMID- 14765938 TI - Should carotid endarterectomy be performed for symptomatic carotid stenosis in Pakistan? AB - OBJECTIVE: The risk of stroke and death associated with carotid endarterctomy is operator dependant. Data regarding risks of this procedure are not available in Pakistan and therefore it is difficult to make accurate risk benefit analysis for individual patients. Our objective was to determine safety of carotid endarterectomy at an academic tertiary care center in Pakistan. METHODS: Patients who underwent carotid endarterectomy (CEA) at our hospital during a ten-year period were identified through ICD-9 coding system of the hospital medical records. Demographic features, associated medical problems and immediate postoperative complications were recorded and analyzed. RESULTS: Sixty-three carotid endarterectomies were performed on 59 patients. Ages range from 43 to 80 (mean 61 +/- 8) years; 53 were male and 10 were female. Common associated diseases among these patients were hypertension; 38 (64.4%), ischemic heart disease; 26 (44%), diabetes mellitus; 24 (40.7%), dyslipidemia; 19 (32.2%) and renal insufficiency; 13 (22%). Most common complication was neuropraxia (transient neuropathy); 5 (7.9%), followed by pneumonia and stroke; each in 3 (4.8%) patients. None of the strokes related to the surgical procedure were disabling. Two of the patients who had stroke, recovered fully within 17 weeks and one recovered partly but was independent in all daily activities of living (ADLs). One patient died following simultaneous coronary artery bypass graft (CABG) and CEA. The risk of stroke or death for patients undergoing CEA was high with simultaneous CABG (3/11, 27%) and low for patients undergoing CEA alone (1/52, 2%). CONCLUSION: Carotid endarterectomy is a safe procedure in patients with symptomatic carotid stenosis at our hospital and should be performed, when indicated. PMID- 14765939 TI - Epidemiology of epilepsy in Pakistan: review of literature. AB - OBJECTIVE: To review literature pertinent to the epidemiology of epilepsy in developing countries with special reference to Pakistan. METHODS: All the studies published in medical journals related to epilepsy in Pakistan were systematically reviewed. Important findings from various studies are summarized. RESULTS: Overall prevalence of epilepsy in Pakistan is estimated to be 9.99 per 1000 population. Highest prevalence is seen in people younger than 30 years of age. A slight decrease in prevalence is noted between the ages of 40 and 59. Higher prevalence is observed in rural population. Etiology of epilepsy is more commonly identified in pediatric population. Epilepsy was considered idiopathic in 21 to 76% cases. Only 27.5% epileptic persons in urban areas and 1.9% in the rural areas were treated with AEDs. The burden of epilepsy is not fully evaluated and understood. Generalized seizures were the most common seizure type noted. Knowledge about epilepsy and its care is extremely low. CONCLUSION: Epilepsy is a common medical problem in Paksitan, more prevalent is rural population. The majority of people with epilepsy are treated inadequately or inappropriately. PMID- 14765940 TI - Status epilepticus in children: a five-year experience at Aga Khan University Hospital. AB - OBJECTIVE: Status epilepticus is an under diagnosed entity in Pakistan. It is a potentially reversible condition but has a high mortality, if it is not recognized and managed on time. The purpose of this study was to determine the clinical profile and the relationship of mortality of status epilepticus with its known risk factors. METHODS: This was a retrospective study. Medical records of all the patients admitted in the last five years (1998-2002) with a diagnosis of status epilepticus (ICDcode 345.30, 345.31) were reviewed. Data was recorded on a Performa and analyzed by using the statistical programme SPSS, chi square and Fischer exact test. RESULTS: The total number of patients were twenty-four. Sixteen patients were males (66.7%). Mean age was fifty-eight months and mean duration of hospital stay 5.5 days (range 2 to 22days). Eight patients were diagnosed to have epilepsy. Four (16.7%) had a previous history of status epilepticus. Three patients presented with status epilepticus for the first time without any previous history of seizures. Ten patients required midazolam infusion (41.7%) and out of these 3 (12.5%) were also given thiopentone infusion to control the seizures. Nine patients were shifted to the ICU for ventilation and control of seizures. Mortality in our study was 25%. Risk factors for mortality included age less than or equal to one year, abnormal MRI, type of the status epilepticus and the total duration of status epilepticus. No significant relationship was found with any of the known risk factors CONCLUSION: Status epilepticus is a neurological emergency. A very high mortality was seen in our study. No risk factors were identified for this high mortality. PMID- 14765941 TI - Clinical practice guidelines for the management of ischemic stroke in Pakistan. PMID- 14765942 TI - Changing patterns of neonatal herpes encephalitis and current treatment guidelines. PMID- 14765943 TI - Critical care myopathy: an emerging medical catastrophe. PMID- 14765944 TI - Disease modification in multiple sclerosis. PMID- 14765945 TI - Leprosy: immunopathology, neurologic manifestations and treatment. PMID- 14765946 TI - Myopathy with Allgrove syndrome. PMID- 14765947 TI - Neuro-imaging facilities in Pakistan. PMID- 14765948 TI - Neurological training in Pakistan. PMID- 14765950 TI - Pursuit of the ineffable: perceptual and motor reversals during the tracking of apparent motion. AB - Pursuit can be guided by perceived rather than physical motion, but the temporal relationship between motion perception and pursuit is unknown. We used an apparent motion stimulus consisting of a horizontal row of evenly spaced Kanizsa illusory squares (1.44 deg2): the illusory contours appeared at the midpoints of the illusory squares presented in the previous frame, producing bi-directional apparent motion of the illusory contours (21.5 deg/s) that could be reversed at will. We measured eye movements in five subjects asked to (1) track the motion of the illusory squares, and (2) reverse the perceived direction while continuing to track the squares. We measured the timing of the voluntary perceptual reversals and compared this to the time course of the reversal in tracking direction. We found that subjects could smoothly track the apparent motion of illusory squares and also produce saccade-free reversals in pursuit velocity. The time course of these motor reversals closely followed the measurements of the perceptual reversal and, on average, the perceptual reversals preceded the pursuit reversals by 53 ms, a delay shorter than when the perceptual reversal was visually guided. Smooth pursuit and the perception of motion direction were in temporal register and highly correlated, suggesting that pursuit can provide a real-time readout for the state of motion perception. PMID- 14765952 TI - Eye-movements aid the control of locomotion. AB - Eye-movements have long been considered a problem when trying to understand the visual control of locomotion. They transform the retinal image from a simple expanding pattern of moving texture elements (pure optic flow), into a complex combination of translation and rotation components (retinal flow). In this article we investigate whether there are measurable advantages to having an active free gaze, over a static gaze or tracking gaze, when steering along a winding path. We also examine patterns of free gaze behavior to determine preferred gaze strategies during active locomotion. Participants were asked to steer along a computer-simulated textured roadway with free gaze, fixed gaze, or gaze tracking the center of the roadway. Deviation of position from the center of the road was recorded along with their point of gaze. It was found that visually tracking the middle of the road produced smaller steering errors than for fixed gaze. Participants performed best at the steering task when allowed to sample naturally from the road ahead with free gaze. There was some variation in the gaze strategies used, but sampling was predominantly of areas proximal to the center of the road. These results diverge from traditional models of flow analysis. PMID- 14765951 TI - Perception can influence the vergence responses associated with open-loop gaze shifts in 3D. AB - We sought to determine if perceived depth can elicit vergence eye movements independent of binocular disparity. A flat surface in the frontal plane appears slanted about a vertical axis when the image in one eye is vertically compressed relative to the image in the other eye: the induced size effect (Ogle, 1938). We show that vergence eye movements accompany horizontal gaze shifts across such surfaces, consistent with the direction of the perceived slant, despite the absence of a horizontal disparity gradient. All images were extinguished during the gaze shifts so that eye movements were executed open-loop. We also used vertical compression of one eye's image to null the perceived slant resulting from prior horizontal compression of that image, and show that this reduces the vergence accompanying horizontal gaze shifts across the surface, even though the horizontal disparity is unchanged. When this last experiment was repeated using vertical expansions in place of the vertical compressions, the perceived slant was increased and so too was the vergence accompanying horizontal gaze shifts, although the horizontal disparity again remained unchanged. We estimate that the perceived depth accounted, on average, for 15-41% of the vergence in our experiments depending on the conditions. PMID- 14765953 TI - Perceived slant from Werner's illusion affects binocular saccadic eye movements. AB - We examined whether binocular saccadic eye movements are determined solely by disparity-defined slant or whether they are influenced by both disparity-defined and perceived slant. The Werner illusion was used to distinguish a plane's disparity-defined slant from its perceived slant. Three subjects viewed a horizontally elongated test strip that was flanked vertically by two planes. The perceived slant of the test strip depended on the slant of the flanking planes. Subjects estimated the perceived slant of the test strip by adjusting the angle between two lines in a symbolic top view. The saccadic eye movements between targets on the test strip were recorded both with visual feedback ("later saccades") and without visual feedback ("first saccades"). We calculated vergence differences for saccades between targets on the test strip (and for fixation on these targets). For each geometrical test strip slant we examined whether the vergence differences could be explained as an effect of perceived slant. This study shows that saccadic eye movements are determined predominantly by the disparity-defined slant, but they can be affected by perceived slant, particularly when multiple saccades are being made. PMID- 14765954 TI - Perceptual and oculomotor evidence of limitations on processing accelerating motion. AB - Psychophysical studies have demonstrated that humans are less sensitive to image acceleration than to image speed (e.g., Gottsdanker, 1956; Werkhoven, Snippe, & Toet, 1992). Because there is evidence that a common motion-processing stage subserves perception and pursuit (e.g., Watamaniuk & Heinen, 1999), either pursuit should be similarly impaired in discriminating acceleration or it must receive input from a system different from the one that processes visual motion for perception. We assessed the sensitivity of pursuit to acceleration or speed, and compared the results with those obtained in perceptual experiments done with similar stimuli and tasks. Specifically, observers pursued or made psychophysical judgments of targets that moved at randomly selected base speeds and subsequent accelerations. Oculomotor and psychophysical discrimination were compared by analyzing performance for the entire stimulus set sorted by either target acceleration or speed. Thresholds for pursuit and perception were higher for target acceleration than speed, further evidence that a common motion-processing stage limits the performance of both systems. PMID- 14765955 TI - Thresholds for stereo-slant discrimination between spatially separated targets are influenced mainly by visual and memory factors but not oculomotor instability. AB - Surface-slant variations can be sensed either simultaneously with steady fixation or sequentially with saccadic gaze shifts. Stereo-slant discrimination thresholds are affected by visual, oculomotor, and memory factors. We have investigated the effects of fixation strategy, target separation, and exposure duration on stereo slant discrimination. With long exposure durations (734 ms), stereo-slant discrimination thresholds measured with simultaneous presentation of test and reference stimuli were lower with gaze shifts than without them when target separations exceeded 4 deg. Above 4-deg target separations, the benefits of improved disparity resolution with foveal gaze shifts outweighed the costs of oculomotor variability associated with saccades. With short exposure durations (167 ms), as target separation increased, stereo-slant discrimination thresholds measured without gaze shifts increased with both sequential and simultaneous stimulus presentations, whereas thresholds with gaze shifts remained constant. This indicates that oculomotor errors are not an important factor in stereo-slant discrimination. In contrast to stereo-slant thresholds, sequential stereo-depth thresholds between two dots, measured with gaze shifts, increased with target separation. Thus, oculomotor error increases with target separation, and it is an important factor in stereo-depth discrimination. PMID- 14765956 TI - Shared motion signals for human perceptual decisions and oculomotor actions. AB - A fundamental question in primate neurobiology is to understand to what extent motor behaviors are driven by shared neural signals that also support conscious perception or by independent subconscious neural signals dedicated to motor control. Although it has clearly been established that cortical areas involved in processing visual motion support both perception and smooth pursuit eye movements, it remains unknown whether the same or different sets of neurons within these structures perform these two functions. Examination of the trial-by trial variation in human perceptual and pursuit responses during a simultaneous psychophysical and oculomotor task reveals that the direction signals for pursuit and perception are not only similar on average but also co-vary on a trial-by trial basis, even when performance is at or near chance and the decisions are determined largely by neural noise. We conclude that the neural signal encoding the direction of target motion that drives steady-state pursuit and supports concurrent perceptual judgments emanates from a shared ensemble of cortical neurons. PMID- 14765957 TI - The contribution of vergence change to the measurement of relative disparity. AB - The relative disparity between two objects in a scene can in principle be measured directly from the retinal images, without knowledge of eye position. But relative disparity increment thresholds are lowest when the relative disparity is small and the objects are not widely separated in the visual field: thus, some relative disparities are easier for the visual system to measure than others. We consider, after others, a second method by which the visual system could measure relative disparity, based on change in vergence ("delta vergence" or DV). The DV mechanism could be more reliable than the retinal mechanism when visual targets are widely separated in visual direction or depth. We used a cue-conflict paradigm to measure the extent to which perceived depth depends on DV. As target separation increased, so did reliance on DV. As intertarget disparity increased, reliance on DV increased for one observer but not for two others. PMID- 14765958 TI - Differential effects of the Muller-Lyer illusion on reflexive and voluntary saccades. AB - Research has produced conflicting evidence as to whether saccade programming is or is not biased by perceptual illusions. However, previous studies have generally not distinguished between effects of illusory percepts on reflexive saccades, programmed automatically in response to an external visual signal, and voluntary saccades, programmed purposively to a location where no signal has occurred. Here we find that voluntary and reflexive saccades are differentially susceptible to the Muller-Lyer illusion; reflexive movements are reliably but modestly affected by the illusion, whereas voluntary movements show an effect similar to that of perceptual judgments. Results suggest that voluntary saccade programming occurs within a non-retinotopic spatial representation similar to that of visual consciousness, whereas reflexive saccade programming occurs within a representation integrating retinotopic and higher level spatial frames. The effects of the illusion on reflexive saccades are not subject to endogenous control, nor are they modulated by the strength of an exogenous target signal. PMID- 14765959 TI - Smooth anticipatory eye movements alter the memorized position of flashed targets. AB - Briefly flashed visual stimuli presented during smooth object- or self-motion are systematically mislocalized. This phenomenon is called the "flash-lag effect" (Nijhawan, 1994). All previous studies had one common characteristic, the subject's sense of motion. Here we asked whether motion perception is a necessary condition for the flash-lag effect to occur. In our first experiment, we briefly flashed a target during smooth anticipatory eye movements in darkness and subjects had to orient their gaze toward the perceived flash position. Subjects reported to have no sense of eye motion during anticipatory movements. In our second experiment, subjects had to adjust a cursor on the perceived position of the flash. As a result, we show that gaze orientation reflects the actual perceived flash position. Furthermore, a flash-lag effect is present despite the absence of motion perception. Moreover, the time course of gaze orientation shows that the flash-lag effect appeared immediately after the egocentric to allocentric reference frame transformation. PMID- 14765960 TI - The extra-retinal motion aftereffect. AB - Repetitive eye movements are known to produce motion aftereffect (MAE) when made to track a moving stimulus. Explanations typically centre on the retinal motion created in the peripheral visual field by the eye movement. This retinal motion is thought to induce perceived motion in the central test, either through the interaction between peripheral MAE and central target or by adaptation of mechanisms sensitive to the relative motion created between centre and surround. Less attention has been paid to possible extra-retinal contributions to MAE following eye movement. Prolonged eye movement leads to afternystagmus which must be suppressed in order to fixate the stationary test. Chaudhuri (1991, Vision Research, 131, 1639-1645) proposed that nystagmus-suppression gives rise to an extra-retinal motion signal that is incorrectly interpreted as movement of the target. Chaudhuri's demonstration of extra-retinal MAE depended on repeated pursuit to induce the aftereffect. Here we describe conditions for an extra retinal MAE that follows more reflexive, nystagmus-like eye movement. The MAE is extra-retinal in origin because it occurs in part of the visual field that received no retinal motion stimulation during adaptation. In an explicit test of the nystagmus-suppression hypothesis, we find extra-retinal MAE fails to store over a 30s delay between adaptation and test. Implications for our understanding of motion aftereffects are discussed. PMID- 14765961 TI - Eye movements facilitate stereo-slant discrimination when horizontal disparity is noisy. AB - Conditions in which saccadic gaze shifts within planar surfaces facilitate stereo slant discrimination for slant about the horizontal and vertical axis were investigated. When horizontal disparity noise was added, large gaze shifts in the direction of the slant lowered stereo-slant discrimination thresholds compared to thresholds measured with steady central fixation, whereas eye movements orthogonal to the slant orientation did not lower slant-discrimination thresholds. When no horizontal noise was added, performance was the same with and without gaze shifts. These results suggest that slant is recovered from depth differences between target edges when horizontal disparity signals are variable and that foveal fixation improves the measures of disparity. Eye movements did not lower slant thresholds by providing multiple foveal samples of slant at different target locations that were averaged to reduce disparity noise levels, because eye movements only lowered the thresholds when there was a depth difference between the fixation points. To study which signals for azimuth are used when slant is recovered from the difference in depth between target edges, vertical disparity noise was added and stimulus height was reduced. Both methods elevated slant-discrimination thresholds when horizontal disparity noise was present, suggesting that vertical disparity is used as a cue for azimuth. PMID- 14765962 TI - The consistency of bisection judgments in visual grasp space. AB - We study whether bisection in visual grasp space (the region over which eye and hand can work together to grasp or touch objects) depends on fixation or on the method of judgment employed (the task). We determined observer bias and sensitivity for bisection judgments (in a fronto-parallel plane as well as along contours slanted in depth). Significant biases were found that varied across observers both qualitatively and quantitatively. These biases were stable for a given individual (across a year between data collection intervals) and across tasks (method of adjustment vs. forced-choice). When observers maintained fixation (on an endpoint or in the neighborhood of the bisection point), fixation location had a small but significant effect on bias, although those effects were small compared with bisection uncertainty. We conclude that bisection judgments differ significantly between fixations, but that the effect of fixation location on bisection is not large enough to be detected reliably by the observer moving his or her eyes during a judgment. PMID- 14765963 TI - The reentry hypothesis: linking eye movements to visual perception. AB - Cortical organization of vision appears to be divided into perception and action. Models of vision have generally assumed that eye movements serve to select a scene for perception, so action and perception are sequential processes. We suggest a less distinct separation. According to our model, occulomotor areas responsible for planning an eye movement, such as the frontal eye field, influence perception prior to the eye movement. The activity reflecting the planning of an eye movement reenters the ventral pathway and sensitizes all cells within the movement field so the planned action determines perception. We demonstrate the performance of the computational model in a visual search task that demands an eye movement toward a target. PMID- 14765964 TI - Task demands and binocular eye movements. AB - Humans make rapid movements of their eyes several times a second that enable them to examine objects located at different positions in space with both of their eyes. Much of our understanding of these binocular movements comes from studies using experienced observers performing repetitive, unnatural tasks. But what eye movements are made when naive observers perform tasks demanding specific binocular visual information? We examined the binocular eye movements produced by observers performing two tasks differing in the visual information needed for their completion. Our motivation for doing this was to examine the role and function of binocular eye movements when making decisions. We considered the fixation strategies adopted by observers, the effects of the task on the dynamics of saccadic eye movements, and the combination of vergence and version in gaze shifts. We report that the task-dependent use of visual information can have a strong influence on the patterns of fixations, whilst not influencing saccade dynamics. Our data provide some support for the notion that observers choose and fixate a notional reference point in the scene when making judgments about depth structure. PMID- 14765965 TI - Human discrimination of visual direction of motion with and without smooth pursuit eye movements. AB - It has long been known that ocular pursuit of a moving target has a major influence on its perceived speed (Aubert, 1886; Fleischl, 1882). However, little is known about the effect of smooth pursuit on the perception of target direction. Here we compare the precision of human visual-direction judgments under two oculomotor conditions (pursuit vs. fixation). We also examine the impact of stimulus duration (200 ms vs. ~800 ms) and absolute direction (cardinal vs. oblique). Our main finding is that direction discrimination thresholds in the fixation and pursuit conditions are indistinguishable. Furthermore, the two oculomotor conditions showed oblique effects of similar magnitudes. These data suggest that the neural direction signals supporting perception are the same with or without pursuit, despite remarkably different retinal stimulation. During fixation, the stimulus information is restricted to large, purely peripheral retinal motion, while during steady-state pursuit, the stimulus information consists of small, unreliable foveal retinal motion and a large efference-copy signal. A parsimonious explanation of our findings is that the signal limiting the precision of direction judgments is a neural estimate of target motion in head-centered (or world-centered) coordinates (i.e., a combined retinal and eye motion signal) as found in the medial superior temporal area (MST), and not simply an estimate of retinal motion as found in the middle temporal area (MT). PMID- 14765966 TI - Depth from motion parallax scales with eye movement gain. AB - Recent findings suggest that the slow eye movement system, the optokinetic response (OKR) in particular, provides the extra-retinal signal required for the perception of depth from motion parallax (Nawrot, 2003). Considering that both the perception of depth from motion parallax (Ono, Rivest & Ono, 1986; Rivest, Ono & Saida, 1989) and the eye movements made in response to head translations (Schwarz & Miles 1991; Paige, Telford, Seidmen, & Barnes, 1998) appear to scale with viewing distance, changes in perceived depth from motion parallax were studied as a function of viewing distance. If OKR is used in the perception of depth from motion parallax, a change in the OKR signal, caused by a change in viewing distance, should accompany a change in perceived depth from motion parallax. Over a range of viewing distances, binocular stereopsis was used to index perceived depth from motion parallax. At these viewing distances the gain of the OKR portion of the compensatory eye movement was also determined. The results show that the change in OKR gain is mirrored by the change in perceived depth from motion parallax as viewing distance increases. This suggests that the OKR eye movement signal serves an important function in the perception of depth from motion. PMID- 14765967 TI - Contributions of fixational eye movements to the discrimination of briefly presented stimuli. AB - Although it is known that images tend to disappear when they are stabilized on the retina for tens of seconds or minutes, the possible functions of fixational movements during the brief periods of visual fixation that occur during natural viewing remain controversial. Studies that investigated the retinal stabilization of stimuli presented for less than a few seconds have observed neither decrement in contrast sensitivity nor image fading. In this study, we analyzed the effect of retinal stabilization on discriminating the orientation of a low-contrast and noisy small bar that was displayed for either 500 ms or 2 s. The bar was randomly tilted by 45 degrees either clockwise or counterclockwise. For both exposure durations, percentages of correct discrimination were significantly lower under conditions of visual stabilization than in the presence of the normally moving retinal image. These results are consistent with the predictions of recent computational models that simulated neuronal responses in the early visual system during oculomotor activity and support the hypothesis that visual processes deteriorate rapidly in the absence of retinal image motion. PMID- 14765968 TI - A comparison of pursuit eye movement and perceptual performance in speed discrimination. AB - Currently there is considerable debate as to the nature of the pathways that are responsible for the perception and motor performance. We have studied the relationship between perceived speed, which is the experiential representation of a moving stimulus, and the speed of smooth pursuit eye movements, the motor action. We determined psychophysical thresholds for detecting small perturbations in the speed of moving patterns, and then by an ideal observer analysis computed analogous "oculometric" thresholds from the eye movement traces elicited by the same stimuli on the same trials. Our results confirm those of previous studies that show a remarkable agreement between perceptual judgments for speed discrimination and the fine gradations in eye movement speed. We analyzed the initial pursuit period of long duration (1000 ms) and short (200 ms) duration perturbations. When we compared the errors for perception and pursuit on a trial by-trial basis there was no correlation between perceptual errors and eye movement errors. The observation that both oculometric and psychometric performance were similar, with Weber fractions in the normal range, but that there is no correlation in the errors suggests that the motor system and perception share the same constraints in their analysis of motion signals, but act independently and have different noise sources. We simulated noise in two models of perceptual and eye movement performance. In the first model we postulate an initial common source for the perceptual and eye movement signals. In that case about ten times the observed noise is required to produce no correlation in trial-by-trial performance. In the second model we postulate that the perceptual signal is a combination of a reafferent eye velocity signal plus the perturbation signal while the pursuit signal is derived from the oculomotor plant plus the perturbation signal. In this model about three times the noise level in the independent signals will mask any correlation due to the common perturbation signal. PMID- 14765969 TI - Saccade target selection in visual search: accuracy improves when more distractors are present. AB - We report four experiments with search displays of Gabor patches. Our aim was to study the accuracy of gaze control in search tasks. In Experiment 1, a target was presented with a single distractor Gabor of a different spatial frequency on the same axis. Subjects could locate the target with the first saccade if the distractor was more distant, but when the distractor was between the fixation point and the target, the first saccade landed much closer to the distractor. In Experiment 2, the number of display items was increased to 16 in a double ring configuration. With this configuration, first saccades were accurately directed to the target, even when there was an intervening distractor in exactly the same configuration as in Experiment 1. Experiment 3 suggested that the improvement in accuracy was not due to distractor homogeneity but rather may be attributable to the increased first saccade latency with the ring configuration. In the final experiment, latency was shown to covary with saccade accuracy. The results are related to a general framework whereby the presence of distractors operates to hold fixation for a longer period of time, thus allowing a greater period of visual processing and more accurate eye movements. PMID- 14765970 TI - The distribution of visual objects on the retina: connecting eye movements and cone distributions. AB - Experimental data on the accuracy and frequency of saccades are incorporated into a model of the visual world and eye movements to determine the spatial distribution of visual objects on the retina. Visual scenes are represented as sequences of discrete small objects whose positions are initially uniformly distributed and then moved toward the center of the retina by eye movements. We then use this model to investigate whether the distribution of cones in the retina maximizes the information transferred about object position. Assuming for simplicity that a single cone is activated by the object, the rate of information transfer is maximized at the receptor stage if the probability that a target lies at a position on the retina is proportional to the local cone density. Although qualitatively it is easy to understand why the cone density is higher at the fovea, by linking the cone density with eye movements through information sampling theory, we provide an explanation for its quantitative variation across the retina. The human cone distribution and the object distribution in our model visual world are shown to have the same general form and are in close agreement between 5- and 30-deg eccentricity. PMID- 14765971 TI - Expansion of visual space after saccadic eye movements. AB - Human subjects reported the perceived two-dimensional location of a visual target that was briefly presented after a saccade in the absence of visual references. Consistent with previous studies, immediately after horizontal saccades, there was a salient horizontal component in mislocalization in the direction opposite to the saccade. However, the horizontal component in mislocalization was not constant and was larger for targets presented further into the visual field contraversive to the saccade. For the same horizontal saccades, the vertical component in mislocalization was also obvious, and it was larger for targets located further away from the saccade trajectory. The saccadic effects resulted in an overall pattern of mislocalization that could be best described as a two dimensional expansion of visual space. The point of expansive origin was not associated with the saccade goal, but was shifted from the saccade goal in the direction of the saccade. These results suggest that spatial information processing at the time of saccades reflects topographic interactions between neural activations from saccade execution and the visual target. The configuration of mislocalized positions of single point stimuli along a line was not comparable to the pattern of non-veridical motion perception described by Park, Lee & Lee (2001), indicating that spatial mislocalization and non-veridical motion perception after saccades are independent phenomena. PMID- 14765973 TI - The interleukin-1 system and female reproduction. AB - Interleukins (ILs) are known best for their involvement in the immune system and their role during inflammation. In the ovary, a growing body of evidence suggests that the ovarian follicle is a site of inflammatory reactions. Thus ovarian cells could represent sources and targets of ILs. Since then, the IL-1 system components (IL-1alpha, IL-1beta, IL-1 receptor antagonist, IL-1 receptors) have been demonstrated to have several sites of synthesis in the ovary. These factors have been localized in the various ovarian cell types, such as the oocyte, granulosa and theca cells, in several mammalian species. IL-1-like bioactivity has been reported in human and porcine follicular fluid at the time of ovulation. The role of IL-1 in local processes is still poorly known, although there is evidence for involvement in the ovulation process, and in oocyte maturation. More precisely, IL-1 may be involved in several ovulation-associated events such as the synthesis of proteases, regulation of plasminogen activator activity, prostaglandin and nitric oxide production. IL-1 also regulates ovarian steroidogenesis. These different aspects of the involvement of the IL-1 system in important aspects of female reproduction are discussed. PMID- 14765974 TI - Heterogenous expression of nestin in human pancreatic tissue precludes its use as an islet precursor marker. AB - The discovery of a pancreatic adult stem cell would have significant implications for cell-based replacement therapies for type 1 diabetes mellitus. Nestin, a marker for neural precursor cells, has been suggested as a possible marker for islet progenitor cells. We have characterized the expression and localization of nestin in both the intact human pancreas and clinical human pancreatic islet grafts. Nestin was found to be expressed at different levels in the acinar component of human pancreatic biopsies depending on donor, as well as in ductal structures and islets to some degree. In islets, insulin-producing beta-cells rarely co-expressed the protein, and in the ducts a small percentage (1-2%) of cells co-expressed nestin and cytokeratin 19 (CK19) while most expressed only CK19 (90%) or nestin (5-10%) alone. Assessment of nestin expression in neonatal pancreatic sections revealed an increased number of islet-associated positive cells as compared with adult islets. Nestin immunoreactivity was also found in cells of the pancreatic vasculature and mesenchyme as evidenced by co localization with smooth muscle actin and vimentin. Samples from post-islet isolation clinical islet grafts revealed a pronounced heterogeneity in the proportion of nestin-positive cells (<1-72%). Co-localization studies in these grafts showed that nestin is not co-expressed in endocrine cells and rarely (<5%) with cytokeratin-positive ductal cells. However, relatively high levels of co expression were found with acinar cells and cells expressing the mesenchymal marker vimentin. In conclusion we have shown a diffuse and variable expression of nestin in human pancreas that may be due to a number of different processes, including post-mortem tissue remodeling and cellular differentiation. For this reason nestin may not be a suitable marker solely for the identification of endocrine precursor cells in the pancreas. PMID- 14765975 TI - Zinc partitions IGFs from soluble IGF binding proteins (IGFBP)-5, but not soluble IGFBP-4, to myoblast IGF type 1 receptors. AB - Zinc (Zn(2+)), a multifunctional micronutrient, was recently shown to lower the affinity of cell-associated insulin-like growth factor (IGF) binding protein (IGFBP)-3 and IGFBP-5 for both IGF-I and IGF-II, but to increase the affinity of the cell surface type 1 IGF receptor (IGF-1R) for the same two ligands. However, there is a need for data concerning the effects of Zn(2+) on soluble IGFBPs and the type 2 IGF receptor (IGF-2R). In the current work, we demonstrate that Zn(2+) affects the affinity of IGFBP-5 secreted by myoblasts but not IGFBP-4. Zn(2+), at physiological levels, depressed binding of both IGF-I and IGF-II to IGFBP-5, affecting (125)I-IGF-I more than (125)I-IGF-II. Both (125)I-IGF-I and (125)I-IGF II bound to high and low affinity sites on IGFBP-5. Zn(2+) converted the high affinity binding sites of IGFBP-5 into low affinity binding sites. An IGF-I analog, (125)I-R(3)-IGF-I, did not bind to the soluble murine IGFBP-5. Zn(2+) also decreased the affinity of the IGF-2R on L6 myoblasts. In contrast, Zn(2+) increased IGF-I, IGF-II and R(3)-IGF-I binding to the IGF-1R by increasing ligand binding affinity on both P(2)A(2a)-LISN and L6 myoblasts. Soluble IGFBP-5 and IGFBP-4 depressed the binding of (125)I-IGF-I and (125)I-IGF-II to the IGF-1R, but did not affect binding of (125)I-R(3)-IGF-I. By depressing the association of the IGFs with soluble IGFBP-5, Zn(2+) partitioned (125)I-IGF-I and (125)I-IGF-II from soluble IGFBP-5 onto cell surface IGF-1Rs. This effect is not seen when soluble L6-derived IGFBP-4 is present in extracellular fluids. We introduce a novel mechanism by which the trace micronutrient Zn(2+) may alter IGF distribution, i.e. Zn(2+) acts to increase IGF-1R binding at the expense of IGF binding to soluble IGFBP-5 and the IGF-2R. PMID- 14765976 TI - Evidence supporting dual, IGF-I-independent and IGF-I-dependent, roles for GH in promoting longitudinal bone growth. AB - The possibility that growth hormone (GH) has effects on long bone growth independent of insulin-like growth factor-I (IGF-I) has long been debated. If this is true, then long bone growth should be more profoundly affected by the absence of GH (since both GH and GH-stimulated IGF-I effects are absent) than by the absence of IGF-I alone (since GH is still present and actually elevated). To test this hypothesis, we compared long bone growth in mice with targeted deletions of Igf1 vs growth hormone receptor (Ghr). Tibial linear growth rate was reduced by approximately 35% in Igf1 null mice and by about 65% in Ghr null mice between postnatal days 20 and 40, a time of peak GH effect during normal longitudinal growth. The Igf1 null mouse growth plate demonstrated significant enlargement of the germinal zone; chondrocyte proliferation and numbers were normal but chondrocyte hypertrophy was significantly reduced. In contrast, the Ghr null mouse germinal zone was hypoplastic, chondrocyte proliferation and numbers were significantly reduced, and chondrocyte hypertrophy was also reduced. We have previously demonstrated that IGF-II is highly expressed in growth plate germinal and proliferative zones, so we considered the possibility that GH stimulated IGF-II production might promote germinal zone expansion and maintain normal proliferation in the Igf1 null mouse growth plate. Supporting this view, IGF-II mRNA was increased in the Igf1 null mouse and decreased in the Ghr null mouse growth plate.Thus, in the complete absence of IGF-I but in the presence of elevated GH in the Igf1 null mouse, reduction in chondrocyte hypertrophy appears to be the major defect in longitudinal bone growth. In the complete absence of a GH effect in the Ghr null mouse, however, both chondrocyte generation and hypertrophy are compromised, leading to a compound deficit in long bone growth. These observations support dual roles for GH in promoting longitudinal bone growth: an IGF-I-independent role in growth plate chondrocyte generation and an IGF-I-dependent role in promoting chondrocyte hypertrophy. The question of whether GH has direct effects on chondrocyte generation is still not settled, however, since it now appears that IGF-II may medicate some of these effects on the growth plate. PMID- 14765977 TI - FSH activates phosphatidylinositol 3-kinase/protein kinase B signaling pathway in 20-day-old Sertoli cells independently of IGF-I. AB - The gonadotropin FSH plays a key role in the control of Sertoli cell function. The FSH molecular mechanism of action is best recognized for its stimulation of the adenylyl cyclase/cAMP pathway. However, other signaling events have also been demonstrated in Sertoli cells. We have recently presented evidence that FSH can stimulate the phosphatidylinositol 3-kinase/protein kinase B (PI3K/PKB) pathway in 20-day-old Sertoli cells. At the same time, it was proposed that in 8-day-old Sertoli cells the effects of FSH on phosphorylated PKB (P-PKB) levels can be explained by a combination of increased secretion of endogenous IGF-I, decreased IGF-binding protein-3 (IGFBP-3) production, and a synergistic action of FSH on IGF-I-dependent PI3K activation. The aim of the present study was to determine whether the effect of FSH on 20-day-old Sertoli cells is mediated by IGF-I secretion. Twenty-day-old rat Sertoli cell cultures were used. FSH stimulation produced a time-dependent increment in P-PKB levels reaching maximal values in 60 min incubations. IGF-I stimulation was also time-dependent reaching maximal values in 15-min incubations. On the other hand, stimulation of the cultures with FSH showed time-dependent inhibition in phosphorylated mitogen-activated protein kinase (P-MAPK) levels. In sharp contrast, stimulation of the cultures with IGF-I showed time-dependent increments in P-MAPK levels reaching maximal stimulus in 15 min incubations. In order to rule out an IGF-I action on FSH stimulation of P-PKB levels, the effect of a specific IGF-I antibody on the ability of both hormones to increase P-PKB levels was evaluated. As expected, the antibody inhibited IGF-I stimulation of P-PKB levels. However, simultaneous addition of an IGF-I antibody with FSH did not modify the ability of the hormone to increase P-PKB levels. The next set of experiments intended to analyze the relevance of a PI3K/PKB pathway to two biological responses of Sertoli cells to FSH and IGF-I. The PI3K inhibitor, wortmannin, dose-dependently decreased FSH-stimulated lactate and transferrin production. On the other hand, wortmannin was not able to modify the ability of IGF-I to stimulate these metabolic events. In addition, the analysis of the participation of a MAPK pathway in IGF-I regulation of Sertoli cell biological responses showed that the MAPK kinase inhibitors, PD98059 and U0126, decreased IGF-I-stimulated transferrin secretion while not modifying IGF-I stimulated lactate levels. In summary, results obtained so far support the hypothesis that FSH action on P-PKB levels and Sertoli cell metabolism in 20-day old animals is not mediated by autocrine regulation of an IGF-I/ IGFBP-3 axis as previously proposed in 8-day-old Sertoli cells. PMID- 14765978 TI - Umbilical cord ghrelin concentrations in small- and appropriate-for-gestational age newborn infants: relationship to anthropometric markers. AB - Ghrelin is a newly discovered orexigenic peptide originating from the stomach. Circulating ghrelin levels reflect acute and chronic energy balance in humans. However, it is not known whether ghrelin also plays a role in energy homeostasis during fetal life. Forty-one small-for-gestational age (SGA) and 34 appropriate for-gestational age (AGA) infants were studied in order to determine whether cord blood ghrelin concentrations were different in SGA infants compared with AGA infants and the relationship to anthropometric measurements at delivery. The cord blood ghrelin concentrations of SGA infants (means+/-S.E.M.; 15.20+/-3.08 ng/ml) were significantly greater than of AGA infants (2.19+/-0.24 ng/ml) (P<0.0001). They were negatively correlated with the infants' birth weights (r=-0.481, P<0.0001) and with body mass index values (r=-0.363, P<0.001). The higher ghrelin concentrations were found in female infants (20.42+/-4.55 ng/ml) than in males (7.05+/-2.27 ng/ml) in the SGA group (P=0.042). These data provide the first evidence that cord ghrelin levels of SGA infants are greater than those of AGA infants and it is suggested that ghrelin is also affected by nutritional status in the intrauterine period. PMID- 14765979 TI - Vagal stimulation exaggerates the inhibitory ghrelin response to oral fat in humans. AB - Ghrelin, the growth hormone secretagogue receptor ligand, is a key regulator of adiposity and food intake. However, the regulation of ghrelin in response to dietary fat intake remains largely unclear. Furthermore, cephalic elevation of ghrelin may influence fat absorption and postprandial lipaemia. Therefore, the aim of this study was to examine the effect of fat ingestion and vagal stimulation on the regulation of plasma ghrelin. Vagal stimulation was achieved by modified sham feeding (MSF). Eight healthy subjects (four male/four female) consumed a 50 g fat load on two separate occasions. On one occasion, the fat load was preceded by the MSF of a meal for 1 h. Blood, appetite and breath were analysed for 5 h postprandially.A 25% (S.E.M. 3.4) suppression in ghrelin concentration was observed after fat ingestion (P<0.001), without an increase in glucose or insulin. MSF in addition to oral fat enhanced ghrelin suppression further, as well as elevating plasma triacylglycerol (P<0.001) and reducing appetite (P<0.001). The fasting ghrelin concentration was inversely correlated with gastric half-emptying time (P=0.036). We conclude that ghrelin release may be influenced directly by both vagal stimulation and oral fat ingestion. PMID- 14765980 TI - Diurnal rhythm of cerebrospinal fluid and plasma leptin levels related to feeding in non-lactating and lactating rats. AB - Leptin suppresses food intake and increases energy expenditure in the hypothalamus. Rats consume most of their daily food intake during the dark phase of the diurnal cycle. Lactating rats have increased food intake, but the involvement of leptin in the regulation of food intake in this physiological condition is not well understood. The present experiment was carried out to determine the circadian pattern of leptin concentrations in plasma and cerebrospinal fluid (CSF) in relation to the feeding behavior of non-lactating and lactating rats. Female rats were maintained on a controlled lighting schedule (lights on between 0600 and 1800 h) and the food intake of lactating rats was two or threefold higher than that of non-lactating rats. In both groups, food intake was three times greater in the dark phase (P<0.01) compared with the light phase. The plasma concentrations of leptin were lower (P<0.01) in lactating rats than non-lactating rats in both light and dark phases, but there were no differences in plasma leptin levels between light and dark phases. In contrast, and in both groups, the leptin concentrations in CSF were lower (P<0.01) in the dark phase than in the light phase. Leptin levels in CSF were lower (P<0.01) in lactating rats than in non-lactating rats. We conclude that a diurnal pattern of leptin levels within the brain (but not in plasma) reflects characteristics of feeding behavior in lactating and non-lactating rats. PMID- 14765981 TI - Progesterone induction of calcitonin expression in the murine mammary gland. AB - Progesterone, via its nuclear receptor, is mandatory not only for the induction and specification of mammary gland ductal side-branching and lobuloalveologenesis but also for carcinogen-induced mammary tumorigenesis. Notwithstanding these recent advances, a more comprehensive molecular explanation of progesterone induced mammary morphogenesis is contingent upon the identification and characterization of mammary molecular targets that are responsive to the progesterone signal. Toward this goal, we report that calcitonin, a 32 amino acid peptide hormone involved in calcium homeostasis, is exclusively expressed in, and secreted from, luminal epithelial cells within the mammary gland of the pregnant mouse, and, importantly, its expression is progesterone-dependent. Conversely, the calcitonin receptor is present during all stages of post-natal mammary development examined, is localized to the myoepithelial cell lineage, and is not regulated by progesterone. Because calcitonin induction spatiotemporally correlates with increases in progesterone-induced mammary gland proliferation and structural remodeling, we posit that calcitonin - through its receptor - may be involved in one or both of these progesterone-dependent processes. PMID- 14765982 TI - Hypothalamic-pituitary-adrenal axis up-regulation in rats submitted to pituitary stalk compression. AB - The present study investigated the hypothalamic-pituitary-adrenal (HPA) axis activity in response to stress in adult male rats submitted to pituitary stalk compression (PSC) or sham operation. Animals received water or oral salt loading (2% NaCl) for one or eight days before the day of the experiment. On the 14th day post-surgery rats were killed under basal conditions or after 15 min immobilization stress. In the PSC group urine output increased significantly and plasma vasopressin (AVP) levels failed to respond to osmotic stimuli. Short-term salt load induced a significant increase in AVP levels in the sham-operated group. The PSC group presented higher adrenocorticotrophin (ACTH) and corticosterone levels compared with sham-operated rats, both in water intake and salt load conditions. Immobilization stress induced a similar increase in plasma ACTH and corticosterone concentrations in sham-operated and PSC groups under water intake. However, long-term salt load blunted the ACTH and corticosterone responses to immobilization stress in sham-operated rats. PSC rats submitted to short- and long-term salt loading presented no changes in ACTH and corticosterone levels after immobilization. Immobilization stress caused neither AVP responses nor plasma osmolality changes in sham and PSC groups. There was no difference in median eminence AVP content among all groups. In conclusion, the high ACTH and corticosterone levels found in PSC rats under water intake and salt loading conditions suggest an up-regulation of the HPA axis, with a preserved adaptive mechanism to chronic stress. PMID- 14765983 TI - Co-localisation of prostaglandin endoperoxide synthase and immunoreactive adrenocorticotrophic hormone in ovine foetal pituitary. AB - Previous studies have shown that both an intact hypothalamic-pituitary-adrenal axis and prostaglandin E(2) (PGE(2)) are involved in the timing of parturition in sheep. PGE(2) is known to be synthesised by the placenta but has also been found in the foetal brain and pituitary. We propose that the enzymes necessary for production of PGE(2) are found in the ovine foetal pituitary and may be able to exert an autocrine and/or paracrine influence on corticotropes, resulting in an increased secretion of immunoreactive adrenocorticotrophin (irACTH). Pituitary tissues from foetal sheep, of gestational ages 119-126 days, were examined by immunohistochemistry. Primary antibodies for prostaglandin H synthase-1 and -2 (PGHS-1 and PGHS-2), microsomal prostaglandin endoperoxide synthase (mPGES) and irACTH were used to probe expressed proteins and Alexa-Fluor red- and green fluorescent secondary antibodies were used to visualise the bound primary antibody. Staining for PGHS-1, PGHS-2 and mPGES was found throughout the foetal anterior pituitary. PGHS-1 and mPGES were widely distributed, including but not restricted to corticotropes. PGHS-2 was less widely distributed but occasionally was found in cells adjacent to corticotropes. The results indicate that locally produced prostaglandins may have an influence on the secretion of ACTH, independent of placental PGE(2). PMID- 14765984 TI - Angiotensin II influences ovarian follicle development in the transgenic (mRen 2)27 and Sprague-Dawley rat. AB - There is accumulating evidence that local renin-angiotensin systems (RASs) influence cell growth and organ function in a variety of tissues including the ovary. The first aim of this study was to characterise the cellular location of RAS components in the rat ovary. This was facilitated by the use of the hypertensive transgenic (mRen-2)27 rat which overexpresses renin and angiotensin in extra-renal tissues. Comparisons were made with normal Sprague-Dawley (SD) rats. The second aim was to determine if the upregulated RAS of the transgenic (mRen-2)27 rat and infusion of angiotensin II (ANG II) in SD rats influences follicle number and litter size. Gene expression, immunohistochemical and autoradiographic techniques were used to identify a discrete RAS including ANG II receptors in the ovarian stroma, follicles (particularly atretic) and to a lesser extent corpora lutea. The RAS at these sites was most abundant in homozygous (HMZ) followed by heterozygous (HTZ) (mRen-2)27 rats and then SD rats. Large antral and preovulatory follicles and litter size were reduced in (mRen-2)27 rats. In HMZ (mRen-2)27 rats and SD rats infused with ANG II, angiotensin 1a (AT(1a)) receptor mRNA in the ovarian stroma was lower than control SD rats and was associated with a reduction in large antral and preovulatory follicles. These findings indicate that upregulation of the ovarian RAS in the rat influences follicular development and, potentially, reproductive capacity. PMID- 14765985 TI - GH prevents apoptosis in cardiomyocytes cultured in vitro through a calcineurin dependent mechanism. AB - The use of GH to treat heart failure has received considerable attention in recent years. Although the mechanisms of its beneficial effects are unknown, it has been implicated in the regulation of apoptosis in several cell types, and cardiomyocyte apoptosis is known to occur in heart failure. We therefore decided to investigate whether GH protects cardiomyocytes from apoptosis. Preliminary experiments confirmed the expression of the GH receptor (GHR) gene in primary cultures of neonatal rat cardiomyocytes (PC), the specific binding of GH by HL-1 cardiomyocytes, and the GH-induced activation of GHR and its classical downstream effectors in the latter. That GH prevented the apoptosis of PC cells deprived of serum for 48 h was shown by DNA electrophoresis and by Hoechst staining assays in which GH reduced the percentage of cells undergoing apoptosis. Similarly, the TUNEL-evaluated pro-apoptotic effect of cytosine arabinoside (AraC) on HL-1 cells was almost totally prevented by pre-treatment with GH. Fluorescence-activated cell sorter (FACS) analysis showed apoptosis in 9.7% of HL-1 cells growing in normal medium, 21.1% of those treated with AraC and 13.9% of those treated with AraC+GH, and that GH increased the percentage of AraC-treated cells in the S/G(2)/M phase from 36.9% to 52.8%. GH did not modify IGF-I mRNA levels or IGF-I secretion in HL-1 cells treated with AraC, and the protection afforded by GH against AraC-induced apoptosis in HL-1 cells was not affected by the presence of anti-IGF-I antibodies, but was largely abolished by the calcineurin-inhibiting combination cyclosporin+FK506. GH also reduced AraC-induced phosphorylation of mitogen-activated protein kinase p38 (MAPK p38) in HL-1 cells. In summary, GH protects PC and HL-1 cells from apoptosis. This effect is not mediated by IGF-I and may involve MAPK p38 as well as calcineurin. PMID- 14765986 TI - The effects of the aromatase inhibitor fadrozole hydrochloride on fetuses and uteri in late pregnant rats. AB - It is well known that progesterone and estrogen are essential hormones for maintaining pregnancy in most mammals. Some specific roles of progesterone for the maintenance of pregnancy have been clarified, but the role of estrogen is not well known. This study examines the effects of the aromatase inhibitor, fadrozole hydrochloride (Fad), on fetuses, uterine physical properties and the mRNA expression of the uterine enzymes that are related to collagen metabolism during late pregnancy in rats. Continuous s.c. infusion with 300 micro g/day Fad from day 14 of pregnancy (day 1=the day of sperm detection) reduced the concentration of plasma estradiol-17beta (E(2)), and did not change that of plasma progesterone, compared with controls. The treatment increased the intrauterine pressure and reduced the size and compliance of the uterine tissue framework. It also caused injuries (hematomata on the extremities) in about one-quarter of fetuses by day 20. The collagen content of the uterine ampullae was not changed by the treatment. Uterine mRNA expressions of matrix metalloproteinase-1 (MMP-1), which degrades collagens, and of lysyl oxidase (LO), which is necessary for the formation of intra- and inter-molecular cross-links of collagen, were examined by quantitative RT-PCR. The treatment with Fad had no effect on the expression of MMP-1 mRNA and increased that of LO mRNA. Daily s.c. injection with 0.2 micro g E(2) restored the changes in uterine physical properties and the mRNA expression of LO caused by the Fad treatment, and prevented fetal injury, indicating that the influences of Fad treatment are due to estrogen deficiency but not to toxicological effects of Fad. These results imply that estrogen deficiency during late pregnancy in rats obstructs development of the uterine tissue framework so as to cause fetal injury. It is possible that an increase in the uterine expression of LO gene may be involved in this obstruction. PMID- 14765987 TI - Factors associated with mortality of patients with myxoedema coma: prospective study in 11 cases treated in a single institution. AB - This study was carried out to investigate the clinical and biochemical factors which might be of importance in predicting the outcome of patients with myxoedema coma. Eleven patients (ten female) aged 68.1+/-19.5 years attended our institution over a period of 18 years. Glasgow and APACHE II scores and serum free thyroxine and TSH were measured in all the patients on entry. Patients were selected at random to be treated with two different regimens of l-thyroxine. Four patients died with the mortality rate being 36.4%. The patients in coma at entry had significantly higher mortality rates than those with minor degrees of consciousness (75% vs 14.3% respectively, P=0.04). The surviving patients had significantly higher Glasgow scores than those who died (11.85+/-2.3 vs 5.25+/ 2.2 respectively, P<0.001). Comparison of the mean values of APACHE II scores between the surviving group and those who died was significantly different (18.0+/-2.08 vs 31.5+/-2.08 respectively, P<0.0001). The degree of consciousness, the Glasgow score and the severity of the illness measured by APACHE II score on entry were the main factors that determined the post-treatment outcome of patients with myxoedema coma. PMID- 14765988 TI - Endothelins as local activators of adrenocortical cells. AB - Besides the classical corticotropic hormones, ACTH and angiotensin II, various regulatory peptides produced by the adrenal gland are thought to participate in the control of corticosteroid secretion. Here, we review the evidence that endothelins (ETs) synthesized within the adrenal cortex may act as autocrine and/or paracrine factors to regulate adrenocortical cell activity. The expression of ETs has been detected in normal, hyperplastic and neoplastic adrenocortical cells. The occurrence of ET receptors has been described in the different zones of the cortex. ETs stimulate the secretion of both glucocorticoids and mineralocorticoids, and modulate the proliferation of adrenocortical cells. The effects of ETs on steroidogenic cells are mediated through the activation of various signaling mechanisms including stimulation of phospholipase C, phospholipase A2 and adenylyl cyclase activity, as well as calcium influx through plasma channels. These observations suggest that locally produced ETs may play an important role in the regulation of corticosteroid secretion and in the control of mitogenesis in normal and tumoral adrenocortical cells. PMID- 14765989 TI - Differentially expressed Maf family transcription factors, c-Maf and MafA, activate glucagon and insulin gene expression in pancreatic islet alpha- and beta cells. AB - A basic-leucine zipper transcription factor, MafA, was recently identified as one of the most important transactivators of insulin gene expression. This protein controls the glucose-regulated and pancreatic beta-cell-specific expression of the insulin gene through a cis-regulatory element called RIPE3b/MARE (Maf recognition element). Here, we show that MafA expression is restricted to beta cells of pancreatic islets in vivo and in insulinoma cell lines. We also demonstrate that c-Maf, another member of the Maf family of transcription factors, is expressed in islet alpha-cells and in a glucagonoma cell line (alphaTC1), but not in gamma- and delta-cells. An insulinoma cell line, betaTC6, also expressed c-Maf, albeit at a low level. Chromatin immunoprecipitation assays demonstrated that Maf proteins associate with insulin and glucagon promoters in beta- and alpha-cell lines, respectively. c-Maf protein stimulated glucagon promoter activity in a transient luciferase assay, and activation of the glucagon promoter by c-Maf was more efficient than by the other alpha-cell-enriched transcription factors, Cdx2, Pax6, and Isl-1. Furthermore, inhibition of c-Maf expression in alphaTC1 cells by specific short hairpin RNA resulted in marked reduction of the glucagon promoter activity. Thus, c-Maf and MafA are differentially expressed in alpha- and beta-cells where they regulate glucagon and insulin gene expression, respectively. PMID- 14765990 TI - Expression and signalling characteristics of the corticotrophin-releasing hormone receptors during the implantation phase in the human endometrium. AB - Corticotrophin-releasing hormone (CRH) has been identified in several peripheral tissues, including the female reproductive organs. CRH is expressed in the placenta, myometrium, epithelial endometrium and the endometrial stromal cells at all phases of the menstrual cycle. Similarly, CRH receptors are present in pregnant and non-pregnant myometrium, placenta and endometrium. Putative roles of CRH in the endometrium include involvement in implantation, decidualisation and maintenance of pregnancy. In this study we sought to investigate in detail the CRH receptor repertoire expressed in the human endometrium and their signalling characteristics. Using RT-PCR we were able to demonstrate the expression of CRH receptor 1alpha (CRH-R1alpha) and CRH-R2alpha in the human endometrium. CRH R1beta was present in 40% of endometrial cDNAs examined. No apparent expression of CRH-R2beta, CRH-R2gamma or any other CRH-R1 splice variants was detected. Chemical cross-linking studies with 125I-ovine CRH revealed that the endometrial CRH receptor has a molecular weight of 45 kDa. Using the non-hydrolysable photoreactive analogue [alpha-32P]GTP-azidoanilide and peptide antisera raised against G-protein alpha-subunits, we then studied coupling of endometrial CRH receptors to G proteins. Treatment of endometrial membranes with human CRH (100 nM) increased the labelling of Gq and Gs, but not Gi or Go. These results were supported by experiments in epithelial cells of the non-pregnant human endometrium in the secretory phase which showed that CRH induced increases in both cAMP and inositol trisphosphate levels. These results suggested that CRH may exert multiple effects in the human endometrium via distinct signalling cascades. These events are possibly mediated via different receptor subtypes. PMID- 14765991 TI - Identification and characterization of a cDNA and the gene encoding the mouse ubiquitously expressed glucose-6-phosphatase catalytic subunit-related protein. AB - Glucose-6-phosphatase (G6Pase) catalyzes the final step in the gluconeogenic and glycogenolytic pathways, the hydrolysis of glucose-6-phosphate (G6P) to glucose and phosphate. This paper describes the identification and characterization of a cDNA and the gene encoding the mouse ubiquitously expressed G6Pase catalytic subunit-related protein (UGRP). The open reading frame of this UGRP cDNA encodes a protein (346 amino acids (aa); Mr 38,755) that shares 36% overall identity (56% similarity) with the mouse G6Pase catalytic subunit (357 aa; Mr 40,454). UGRP exhibits a similar predicted transmembrane topology and conservation of many of the catalytically important residues with the G6Pase catalytic subunit; however, unlike the G6Pase catalytic subunit, UGRP does not catalyze G6P hydrolysis and does not contain a carboxy-terminal di-lysine endoplasmic reticulum retention signal. UGRP mRNA was detected by RNA blot analysis in every mouse tissue examined with the highest expression in heart, brain, testis and kidney. Database analysis showed that the mouse UGRP gene is composed of six exons, spans approximately 4.2 kbp of genomic DNA and is located on chromosome 11 along with the G6Pase catalytic subunit gene. The UGRP gene transcription start sites were mapped by primer extension analysis, and the activity of the mouse UGRP gene promoter was analyzed using luciferase fusion gene constructs. In contrast to the G6Pase catalytic subunit gene promoter, the UGRP promoter was highly active in all cell lines examined. PMID- 14765992 TI - Epigenetic regulation of inhibin alpha-subunit gene in prostate cancer cell lines. AB - Inhibin was first identified as a gonad-derived regulator of pituitary FSH; however, it has subsequently been shown to be a tumour suppressor in the gonad and adrenal glands. Whereas non-malignant regions of human primary prostate carcinomas express inhibin alpha-subunit (INHA), malignant tissues lack INHA transcript and protein, which is consistent with epigenetic regulation of the inhibin alpha-subunit gene (INHA) promoter. This study investigated whether methylation of the INHA promoter was responsible for inactivation of INHA transcription and translation in the prostate cancer cell lines, LNCaP, DU145 and PC3. Methylation of the promoter was revealed by bisulphite genomic sequencing and use of inhibitors of methylation and histone deacetylation resulted in reactivation of the INHA transcription and translation. Significant (P<0.05) downregulation of a luciferase reporter gene downstream from a methylated INHA promoter compared with unmethylated INHA promoter occurred in vitro. The data demonstrate that promoter methylation is associated with downregulation of the INHA gene in prostate cancer cell lines, which is consistent with its tumour suppressive role. Therefore INHA has a significant role in prostate tumorigenesis. PMID- 14765993 TI - Ubc9 interacts with chicken ovalbumin upstream promoter-transcription factor I and represses receptor-dependent transcription. AB - Chicken ovalbumin upstream promoter-transcription factors (COUP-TFs) are orphan receptors involved in regulation of neurogenesis and organogenesis. COUP-TF family members are generally considered to be transcriptional repressors and several mechanisms have been proposed to underlie this activity. To explore novel transcriptional coregulators for COUP-TFs, we used the COUP-TFI as bait in a yeast two-hybrid screen of an adrenocortical adenoma cDNA library. We have identified Ubc9, a class E2 conjugating enzyme of small ubiquitin-related modifier (SUMO)-1 as a COUP-TFI corepressor. Ubc9 interacts with COUP-TFI in yeast and in glutathione S-transferase pulldown and coimmunoprecipitation assays. Fluorescence imaging studies show that both Ubc9 and COUP-TFI are colocalized in the nuclei of transfected COS-1 cells. The C-terminal region of Ubc9 encoding amino acids 59-158 interacts with the C-terminus of COUP-TFI encoding amino acids 383-403, in which transcriptional repression domains are located. Mammalian one hybrid assays utilizing a variety of Ubc9 fragments fused to Gal4 DNA-binding domain show that a Ubc9 fragment encoding amino acids 1-89 contains autonomous transferrable repression domain. Transfection of Ubc9 into COS-1 cells markedly enhances transcriptional repression by Gal4 DNA-binding domain-fused to COUP TFI(155-423), but not by Gal4-COUP-TFI(155-388) which lacks a repressor domain. Coexpression of a C-terminal deletion mutant of Ubc9(1-58), which fails to interact with COUP-TFI, but retains a transcriptional repression domain, has no effect on Gal4-COUP-TFI-mediated repression activity. These findings indicate that interaction of Ubc9 with COUP-TFI is crucial for the corepressor function of Ubc9. Overexpression of Ubc9 similarly enhances COUP-TFI-dependent repression of the promoter activity of the bovine CYP17 gene encoding steroid 17alpha hydroxylase. In addition, the C93S mutant of Ubc9, which abrogates SUMO-1 conjugation activity, continues to function as a COUP-TFI corepressor. Our studies indicate that Ubc9 functions as a novel COUP-TFI corepressor, the function of which is distinct from its SUMO-1 conjugating enzyme activity. PMID- 14765994 TI - Calcitonin stimulates expression of the rat 25-hydroxyvitamin D3-24-hydroxylase (CYP24) promoter in HEK-293 cells expressing calcitonin receptor: identification of signaling pathways. AB - Regulation of the gene for renal 25-hydroxyvitamin D-24-hydroxylase (CYP24) is important for controlling the level of circulating 1,25-dihydroxyvitamin D3 (1,25(OH)2D3). We report here for the first time that the peptide hormone calcitonin significantly stimulates expression of a rat CYP24 promoter-luciferase construct in both transiently and stably transfected kidney HEK-293 cells. A GC box at -114/-101 and a CCAAT box at -62/-51 have been identified that underlie both basal expression of the CYP24 promoter and the calcitonin inductive response. Data from overexpression studies suggested that Sp1 and NF-Y are the proteins that function through the GC and CCAAT boxes respectively. ERK1/2 signaling pathways were not involved in the calcitonin-mediated response, since stimulation of the promoter was unaffected by the pharmacological ERK1/2 inhibitor PD98059 and by a dominant negative mutant of ERK1/2 (ERK1K71R). In contrast, calcitonin induction but not basal expression was dependent on protein kinase A and protein kinase C (PKC) activities with the inhibitors H89 and calphostin C lowering induction by 50-60%. The atypical PKC, PKCzeta contributes to calcitonin induction, but not to basal expression of the CYP24 promoter, since overexpression of a dominant negative clone PKCzetaK281 M lowered induction by 50%. Cotransfection of a dominant negative form of Ras resulted in calcitonin mediated induction being reduced also by about 50%. A Ras-PKCzeta signaling pathway for calcitonin action is proposed, which acts through the GC box. The findings have been extrapolated to the in vivo situation where we suggest that induction of renal CYP24 by calcitonin could be important under hypercalcemic conditions thus contributing to the lowering of circulating 1,25(OH)2D3 levels. PMID- 14765995 TI - Functional importance of Myc-associated zinc finger protein for the human parathyroid hormone (PTH)/PTH-related peptide receptor-1 P2 promoter constitutive activity. AB - The aim of the present study was to analyze the functional importance for the parathyroid hormone (PTH)/PTH-related peptide (PTHrP) receptor (PTHR1) gene P2 promoter activity of the putative proximal Myc-associated zinc finger protein (MAZ) site localized at position bp -45 to -39 bp, taking advantage of a G/A mutation identified at position -40 in the human sequence. Wild-type 'full length' (1285P2) and truncated (760P2) promoter sequences were inserted upstream to the luciferase basic (pLucB) and enhancer (pLucE) reporter gene expression vectors. Transient transfections in osteoblast-like SaOS-2 cells and renal cells (RC.SV3A2) showed that the -40 G/A mutation significantly impaired transcriptional activity of wild-type 1285P2-pLucB and 760P2-pLucE promoter constructs. Further truncation of the P2 sequence demonstrated that the sequence 109/-37 bp was essential for promoter activity. Co-transfection with a MAZ expression vector did not modify the wild-type 1285P2-pLucB construct reporter activity but significantly increased 2-fold the mutated construction activity (P<0.05). Electrophoretic mobility shift assays using SaOS-2 nuclear extracts and a double-stranded DNA fragment encompassing the -45 to -39 putative MAZ site (ds MAZ-oligo) disclosed two specific DNA-protein complexes. Complex II (fast moving) had a lower affinity for the mutated MAZ motif than for the wild-type MAZ motif while complex I (slow moving) had the same affinity for both wild-type or mutated MAZ sequences. Competition studies with Sp1 consensus oligonucleotide (ds-Sp1 oligo) markedly reduced complex I intensity, with a concomitant increase in that of complex II. Finally, ribonuclease protection assays showed that P2-specific PTHR1 mRNA transcript expression was significantly decreased in SaOS-2 cells transfected with ds-MAZ-oligo as compared with that for control (P<0.001) and ds Sp1-oligo (P<0.05). Taken together, our studies suggest that the putative -45 to 39 MAZ-binding site regulates the constitutive activity of human PTHR1 P2 promoter. PMID- 14765996 TI - Chorionic gonadotrophin beta subunit mRNA but not luteinising hormone beta subunit mRNA is expressed in the pituitary of the common marmoset (Callithrix jacchus). AB - The pituitary gonadotrophins LH and FSH are responsible for regulation of gametogenesis in the testis and ovary. Chorionic gonadotrophin (CG), a third closely related glycoprotein hormone derived by gene duplication of the LHbeta gene and secreted by the placenta in primates, is essential for the rescue of the corpus luteum and maintenance of pregnancy. We have recently shown that marmoset (m) CGbeta mRNA is highly expressed in the pituitary of the common marmoset (Callithrix jacchus) and that LH is less active than human CG in activating the human LH receptor lacking exon 10. To investigate further which gonadotrophin is the actual ligand of the LH receptor (LHR) of the marmoset monkey that naturally lacks exon 10, we identified and characterised the genomic organisation of the mLHbeta gene and its expression. Intergenic PCR amplification of the region encompassing the mLHbeta and the mCGbeta genes revealed that, surprisingly, mCGbeta is located 20 kbp upstream of the LHbeta gene, whereas in other species the intergenic distance is approximately 2-3 kbp. Sequence analysis of the mLHbeta coding region showed 70% identity to mCGbeta and 90% identity to human LHbeta at the amino acid level. Both gonadotrophin beta subunits are present at the genomic level, but RT-PCR of pituitary and placental total RNA using specific oligonucleotides for mCGbeta and mLHbeta showed high expression of mCGbeta mRNA in both tissues, whereas LHbeta was expressed neither in the pituitary nor in the placenta. Thus mLHbeta mRNA is lacking in the marmoset pituitary. Immunohistochemistry of marmoset pituitaries showed that mCG was confined to the gonadotrophes, and partly co-localised in cells stained positively for FSH. Western blot analysis confirmed the presence of mCG in the pituitary. Northern blot analysis using mCGbeta as a probe displayed one transcript of 0.7 kb in the pituitary and detected two transcripts of 1.1 kb and 2 kb in the marmoset placenta. Our results suggest that, in the common marmoset, CG is the only gonadotrophin with luteinising function that is present in the pituitary. We postulate that, owing to an unknown mutational event in evolution, expression of mLH was completely abolished, and CG - which, unlike LH, acts normally even when exon 10 is missing from the LHR - took over its function. PMID- 14765997 TI - A1 adenosine receptors mediate hypoglycemia-induced neuronal injury. AB - The cellular mechanisms that lead to neuronal death following glucose deprivation are not known, although it is recognized that hypoglycemia can lead to perturbations in intracellular calcium ([Ca2+]i) levels. Recently, activation of A1 adenosine receptors (A1AR) has been shown to alter [Ca2+]i and promote neuronal death. Thus, we examined if A1AR activation contributes to hypoglycemia induced neuronal injury using rat cortical neurons. First, we observed that hypoglycemia was associated with large increases in neuronal adenosine release. Next, decreased neuronal viability was seen with progressive reduction in glucose concentration (25, 6, 3, 0.75 and 0 mM). Using the calcium-sensitive dye, Fluo-3, we observed both acute and long-term changes in relative [Ca2+]i during hypoglycemic conditions. Demonstrating a role for adenosine in this process, both the loss in neuronal viability and the early changes in [Ca2+]i were reversed by treatment with A1AR antagonists (8-cyclopentyl, 1,3-dipropylxanthine; 9-chloro-2 (2-furyl)(1,2,4)-triazolo(1,5-c)quinazolin-5-amine; and N-cyclopentyl-9 methyladenine). We also found that hypoglycemia induced the expression of the pro apoptotic enzyme, caspase-3, and that A1AR antagonism reversed hypoglycemia induced caspase-3 activity. Collectively, these data show that hypoglycemia induces A1ARs activation leading to alterations in [Ca2+]i, which plays a prominent role in leading to hypoglycemia-induced neuronal death. PMID- 14765998 TI - Regulation of adipose tissue leptin secretion by alpha-melanocyte-stimulating hormone and agouti-related protein: further evidence of an interaction between leptin and the melanocortin signalling system. AB - The central role of the melanocortin system in the regulation of energy balance has been studied in great detail. However, the functions of circulating melanocortins and the roles of their peripheral receptors remain to be elucidated. There is increasing evidence of a peripheral action of melanocortins in the regulation of leptin production by adipocytes. Here we investigate the interaction of alpha-melanocyte stimulating hormone (alpha-MSH) and agouti related protein (AgRP) in the regulation of leptin secretion from cultured rat adipocytes and examine the changes in circulating alpha-MSH and AgRP in lean and obese rodents after hormonal and energetic challenge. Leptin secretion (measured by ELISA) and gene expression (by real-time quantitative PCR) of differentiated rat adipocytes cultured in vitro were inhibited by the administration of alpha MSH (EC50=0.24 nM), and this effect was antagonised by antagonists of the melanocortin receptors MC4R and MC3R (AgRP and SHU9119). The presence of MC4R in rat adipocytes (RT-PCR and restriction digest) supports the involvement of this receptor subtype in this interaction. Leptin administered to ob/ob mice in turn increases the release of alpha-MSH into the circulation, suggesting a possible feedback loop between the site of alpha-MSH release and the release of leptin from the adipose tissue. However, the physiological significance of this putative feedback probably depends upon the underlying state of energy balance, since in the fasting state low plasma alpha-MSH is paralleled by low plasma leptin. PMID- 14765999 TI - Plasminogen activator inhibitor-1 promoter activity in adipocytes is not influenced by the 4 G/5 G promoter polymorphism and is regulated by a USF-1/2 binding site immediately preceding the polymorphic region. AB - Plasminogen activator inhibitor-1 (PAI-1) levels were found to be associated with obesity indicating that adipocytes influence PAI-1 plasma levels. In addition, the 4 G/5 G promoter polymorphism of the PAI-1 gene may modulate PAI-1 transcription. We investigated the transcriptional regulation of the human PAI-1 gene in adipocytes and analyzed the genetic contribution of the 4 G/5 G polymorphism. The PAI-1 promoter was analyzed using electrophoretic mobility shift assays (EMSAs) and luciferase reporter gene assays. A putative binding site for the upstream stimulatory factor-1/2 (USF-1/2) at the polymorphic region of the PAI-1 promoter was identified. The binding of USF-1/2 was studied using nuclear extracts prepared from adipocytes and was similar in all the promoter variants as analyzed by EMSA. A 257 bp PAI-1 promoter fragment including the 4 G/5 G site was transcriptionally active in adipocytes and was not influenced by the polymorphism. The present data indicate for the first time that USF-1/2 is transcriptionally active in differentiated adipocytes. However, USF-1/2 binding activity and PAI-1 transcription are not influenced by the 4 G/5 G-allele. These data possibly explain the observation that PAI-1 secretion from adipose tissue is not influenced by the PAI-1 promoter polymorphism. PMID- 14766000 TI - Differential expression of two somatostatin receptor subtype 1 mRNAs in rainbow trout (Oncorhynchus mykiss). AB - Somatostatins (SSs) play important roles in the growth, development and metabolism of vertebrates. In this study, cDNAs for two unique somatostatin receptor variants were cloned and sequenced from rainbow trout. The two cDNAs, one consisting of 1755 bp and the other of 1743 bp, share 63.6% identity in nucleotide sequence and 94.1% identity in deduced amino acid sequence and presumably arose through gene duplication. Each cDNA encodes for a putative 371 amino acid somatostatin receptor (one designated sst1A and the other sst1B) containing seven transmembrane domains. Rainbow trout sst1A and sst1B have 64.4 and 65.5% similarity respectively with human sst1 and only 43-60% similarity with other subtypes. Trout sst1 mRNAs are differentially expressed, both in terms of distribution among tissues as well as in terms of abundance within selected tissues. Both sst1A and sst1B mRNAs were present in brain, stomach, liver, pancreas, upper and lower intestine, pyloric cecum, kidney and muscle, whereas only sst1B mRNA was present in the esophagus. sst1A mRNA was more abundant than sst1B in the optic tectum, whereas sst1B mRNA was more abundant than sst1A in liver. sst1A and sst1B mRNAs were equally abundant in pancreas. These findings contribute to the understanding of the evolution of the SS signaling system and provide insight into the mechanisms that regulate the expression of SS receptors. PMID- 14766001 TI - Cloning of the cDNA for thyroid stimulating hormone beta subunit and changes in activity of the pituitary-thyroid axis during silvering of the Japanese eel, Anguilla japonica. AB - The purposes of this study were: (1) to clone the cDNA encoding pituitary thyroid stimulating hormone beta subunit (TSH beta) of the Japanese eel, Anguilla japonica, together with its genomic DNA sequence, for phylogenetic analysis, and to study the regulation of the TSH beta gene expression in cultured pituitaries; and (2) to investigate the transcript levels of pituitary TSH beta mRNA and the serum thyroxine profiles at different stages of ovarian development before and during silvering in the wild female eels. The maturity of female eels was divided into four stages, juvenile, sub-adult, pre-silver, and silver, based on skin color and oocyte diameter. The genomic DNA of the TSH beta subunit contains two introns and three exons, and the TSH beta protein possesses a putative signal peptide of 20 amino acids and a mature peptide of 127 amino acids. The amino acid sequence identities of TSH beta mature peptide of Japanese eel compared with those of teleosts and other vertebrates are: European eel (98.4%), salmonids (60.6-61.3%), carps (52.0-56.7%), sturgeon (48.4%), and tetrapods (42.9-45.2%). In in vitro studies of the regulation of TSH beta mRNA it was found that thyrotropin-releasing hormone increased while thyroxine decreased its expression. RT-PCR and real-time quantitative PCR analysis showed that the transcript levels of TSH beta subunit increased during eel silvering. The serum thyroxine levels also increased in parallel with TSH beta mRNA expression during silvering, supporting the hypothesis that the hypothalamus-pituitary-thyroid axis is correlated to silvering in the wild female Japanese eels. PMID- 14766002 TI - Differential hepatic gene expression in a polygenic mouse model with insulin resistance and hyperglycemia: evidence for a combined transcriptional dysregulation of gluconeogenesis and fatty acid synthesis. AB - New Zealand obese (NZO) mice exhibit severe insulin resistance of hepatic glucose metabolism. In order to define its biochemical basis, we studied the differential expression of genes involved in hepatic glucose and lipid metabolism by microarray analysis. NZOxF1 (SJLxNZO) backcross mice were generated in order to obtain populations with heterogeneous metabolism but comparable genetic background. In these backcross mice, groups of controls (normoglycemic/normoinsulinemic), insulin-resistant (normoglycemic/hyperinsulinemic) and diabetic (hyperglycemic/hypoinsulinemic) mice were identified. At 22 weeks, mRNA was isolated from liver, converted to cDNA, and used for screening of two types of cDNA arrays (high-density filter arrays and Affymetrix oligonucleotide microarrays). Differential gene expression was ascertained and assessed by Northern blotting. The data indicate that hyperinsulinemia in the NZO mouse is associated with: (i) increased mRNA levels of enzymes involved in lipid synthesis (fatty acid synthase, malic enzyme, stearoyl-CoA desaturase) or fatty acid oxidation (cytochrome P450 4A14, ketoacyl CoA thiolase, acyl-CoA oxidase), (ii) induction of the key glycolytic enzyme pyruvate kinase, and (iii) increased mRNA levels of the gluconeogenic enzyme phosphoenolpyruvate carboxykinase. These effects were enhanced by a high-fat diet. In conclusion, the pattern of gene expression in insulin-resistant NZO mice appears to reflect a dissociation of the effects of insulin on genes involved in glucose and lipid metabolism. The data are consistent with a hypothetical scenario in which an insulin-resistant hepatic glucose production produces hyperinsulinemia, and an enhanced insulin- and substrate-driven lipogenesis further aggravates the deleterious insulin resistance of glucose metabolism. PMID- 14766003 TI - Androgen inhibition of MAP kinase pathway and Elk-1 activation in proliferating osteoblasts. AB - Non-aromatizable androgens have significant beneficial effects on skeletal homeostasis independently of conversion to estradiol, but the effects of androgens on bone cell metabolism and cell proliferation are still poorly understood. Using an osteoblastic model with enhanced androgen responsiveness, MC3T3-E1 cells stably transfected with androgen receptor (AR) under the control of the type I collagen promoter (colAR-MC3T3), the effects of androgens on mitogenic signaling were characterized. Cultures were treated with the non aromatizable androgen 5alpha-dihydrotestosterone (DHT) and the effects on osteoblast viability were determined as measured by an MTT assay. A complex response was observed in that continuous short-term DHT treatment enhanced osteoblast viability, but with longer-term DHT treatment inhibition was observed. The inhibition by DHT was prevented by the specific AR antagonist hydroxyflutamide, and was also observed in primary cultures of normal rat calvarial osteoblasts. In order to identify potential mediators of this effect, mitogenic pathway-specific cDNA microarrays were interrogated. Reduced hybridization of several genes important in MAP kinase-mediated signaling was observed, with the most dramatic effect on Elk-1 expression. Analysis of phosphorylation cascades demonstrated that DHT treatment inhibited phosphoERK1/2 levels, MAP kinase activation of Elk-1, Elk-1 protein and phosphoElk-1 levels, and downstream AP-1/luciferase reporter activity. Together, these data provide the first evidence that androgen inhibition of the MAP kinase signaling pathway is a potential mediator of osteoblast growth, and are consistent with the hypothesis that the MAP cascade may be a specific downstream target of DHT. PMID- 14766004 TI - Evidence that protein kinase Cdelta is not required for palmitate-induced cytotoxicity in BRIN-BD11 beta-cells. AB - Chronic exposure of pancreatic beta-cells to saturated fatty acids leads to loss of viability, an effect that has been implicated in the process of beta-cell 'lipotoxicity' associated with the progression of type 2 diabetes. The mechanisms involved are unknown but recent evidence has implicated the delta isoform of protein kinase C (PKCdelta) in mediating fatty acid toxicity. We have investigated this proposition in the clonal insulin-secreting cell line, BRIN BD11. BRIN-BD11 cells were found to undergo apoptosis when exposed to palmitate and this response was attenuated by the purportedly selective inhibitor of PKCdelta, rottlerin. However, activation of PKCdelta with the phorbol ester, phorbol-12-myristate-13-acetate (PMA), failed to promote cell death and down regulation of PKCdelta did not prevent the cytotoxic effects of palmitate. Moreover, rottlerin remained effective as a blocker of the palmitate response in cells depleted of PKCdelta. Since rottlerin can inhibit various other kinases in addition to PKCdelta, a range of additional kinase inhibitors was also tested. Of these, only the putative Ca2+/calmodulin-dependent protein kinase II (CaM kinase II) inhibitor, KN-62, was found to inhibit palmitate-induced cell death. However, this effect was not reproduced by a more selective pseudo-substrate inhibitor of CaM kinase II. Therefore, the present results reveal that palmitate induces cell death in BRIN-BD11 cells and suggest that this may involve the activation of a rottlerin (and KN-62)-sensitive kinase. However, it is clear that PKCdelta is not required for this response. PMID- 14766005 TI - Postprandial lipemia is modified by the presence of the polymorphism present in the exon 1 variant at the SR-BI gene locus. AB - It has recently been reported that carriers of the less common allele at the scavenger receptor class B type I (SR-BI) exon 1 polymorphism are more susceptible to the presence of saturated fatty acid in the diet because of a greater increase in LDL cholesterol. Our aim was to determine if this polymorphism could also influence postprandial lipoprotein metabolism, because the SR-BI has been described as a possible mediator in the intestinal absorption of triacylglycerols. Forty-seven normolipidemic volunteers who were homozygous for the E3 allele at the APOE gene were selected [37 homozygous for the common genotype (1/1) at the SR-BI exon 1 polymorphism and 10 heterozygous (1/2)]. They were given a fat-rich meal containing 1 g fat and 7 mg cholesterol per kg body weight and vitamin A 60,000 IU/m2 body surface. Fat accounted for 60% of calories, and protein and carbohydrates accounted for 15% and 25% of energy respectively. Blood samples were taken at time 0, every 1 h until 6 h, and every 2.5 h until 11 h. Total cholesterol and triacylglycerols in plasma, and cholesterol, triacylglycerols and retinyl palmitate in triacylglycerol-rich lipoproteins (large and small triacylglycerol-rich lipoproteins) were determined. Postprandial responses for triacylglycerols and retinyl palmitate in small triacylglycerol-rich lipoproteins were higher in 1/1 individuals than in 1/2 individuals. No other significant differences were noted. Our data show that the presence of the genotype 1/2 is associated with a lower postprandial lipemic response. PMID- 14766006 TI - Tightly regulated and inducible expression of a yoked hormone-receptor complex in HEK 293 cells. AB - We have previously reported the construction of a constitutively active luteinizing hormone receptor by covalently linking a fused heterodimeric hormone to the extracellular domain of the G protein-coupled receptor. This yoked hormone receptor complex (YHR) was found to produce high levels of cAMP in the absence of exogenous hormone. Stable lines expressing YHR were generated in HEK 293 cells to obtain lines with different expression levels; however, in a relatively short time of continued passage, it was found that YHR expression was greatly reduced. Herein, we describe the development of clonal lines of HEK 293 cells in which the expression of YHR is under the control of a tetracycline-regulated system. Characterization of clonal lines revealed tight control of YHR expression both by dose and time of incubation with doxycycline. These experiments demonstrated a good correlation between expression levels of the receptor and basal cAMP production. Moreover, the reduction in receptor expression following doxycycline removal revealed that YHR mRNA and protein decayed at similar rates, again suggesting a strong linkage between mRNA and protein levels. The controlled expression of YHR in this cell system will allow for a more detailed analysis of the signaling properties associated with constitutive receptor activation and may prove to be advantageous in developmental studies with transgenic animals. PMID- 14766007 TI - Expressions of hepatic genes, especially IGF-binding protein-1, correlating with serum corticosterone in microarray analysis. AB - Microarray technology was evaluated for usefulness in assessing relationships between serum corticosterone and hepatic gene expression. Nine pairs of female Swiss mice were chosen to provide a wide range of serum corticosterone ratios; cDNA microarray analysis (approximately 8000 genes) was performed on their livers. A statistical method based on calculation of 99% confidence intervals discovered 32 genes which varied significantly among the livers. Five of these ratios correlated significantly with serum corticosterone ratio, including tyrosine aminotransferase, stress-induced protein, pleiotropic regulator 1 and insulin-like growth factor-binding protein-1; the latter has a potential role in cancer development. Secondly, linear regression of gene expression vs corticosterone ratios was screened for those with r> or =0.8 (P<0.01), yielding 141 genes, including some known to be corticosterone regulated and others of interest as possible glucocorticoid targets. Half of these significant correlations involved data sets where no microarray ratio exceeded +/- 1.5. These results showed that microarray may be used to survey tissues for changes in gene expression related to serum hormones, and that even small changes in expression can be of statistical significance in a study with adequate numbers of replicate samples. PMID- 14766008 TI - Human immature germ cells and ejaculated spermatozoa contain aromatase and oestrogen receptors. AB - It is now well established that oestrogens play a part in germ cell function. These hormones are synthesised by the cytochrome P450 aromatase (P450 arom) and act via two kinds of receptor (ERalpha and ERbeta). Although the presence of aromatase and oestrogen receptors in mammalian testis is now well documented, the localisation of these proteins in human germ cells is not yet clear. The primary purpose of the current study was to look for the expression of aromatase and oestrogen receptors in human germ cells. Human immature germ cells were collected from semen samples with an excess of rounds cells (>20%) and purified spermatozoa were obtained after sedimentation on a discontinuous PureSperm gradient. Expression of aromatase and oestrogen receptors was determined by RT-PCR with specific primers, and by Western blot using monoclonal antibodies. RT-PCR products for aromatase, ERalpha and ERbeta were amplified from total RNA isolated from human germ cells and spermatozoa. We identified an ERalpha isoform variant that lacks exon 4 in human germ cells and visualised P450 arom as a single band of 49 kDa in germ cells, as we have already reported for human ejaculated spermatozoa. By Western blot, we identified two proteins for ERbeta at approximately 50 and approximately 60 kDa, which could correspond to the long and short forms of ERbeta formed from the use of alternative start sites. In human ejaculated spermatozoa, ERbeta protein was not detected, even though we could amplify mRNA. Using Western blot analysis and a monoclonal antibody specific for ERalpha, we detected two proteins in human immature germ cells: one of the expected size (66 kDa) and a second one of 46 kDa. In mature spermatozoa, only the 46 kDa band was observed and we speculate it may be related to the ERalpha isoform lacking exon I. In conclusion, we have identified P450 arom and ER proteins (full-length and variant) in human germ cells. Further studies are now required to elucidate the mechanism of action of oestrogens on human male germ cells, in terms of both genomic and 'non-genomic' pathways. PMID- 14766009 TI - The cAMP signaling system regulates LHbeta gene expression: roles of early growth response protein-1, SP1 and steroidogenic factor-1. AB - Expression of the gonadotropin genes has been shown to be modulated by pharmacological or physiological activators of both the protein kinase C (PKC) and the cAMP second messenger signaling pathways. Over the past few years, a substantial amount of progress has been made in the identification and characterization of the transcription factors and cognate cis-elements which mediate the PKC response in the LH beta-subunit (LHbeta) gene. In contrast, little is known regarding the molecular mechanisms which mediate cAMP-mediated regulation of this gene. Using pituitary cell lines, we now demonstrate that rat LHbeta gene promoter activity is stimulated following activation of the cAMP system by the adenylate cyclase activating agent, forskolin, or by the peptide, pituitary adenylate cyclase-activating peptide. The forskolin response was eliminated with mutation of a previously identified 3' cis-acting element for the early growth response protein-1 (Egr-1) when evaluated in the context of region 207/+5 of the LHbeta gene. Activation of the cAMP system increased Egr-1 gene promoter activity, Egr-1 protein levels and Egr-1 binding to the LHbeta gene promoter, supporting the role of this transcription factor in mediating the cAMP response. Analysis of a longer LHbeta promoter construct (-797/+5) revealed additional contribution by upstream Sp1 DNA-regulatory regions. Of interest, forskolin-induced stimulation of LHbeta gene promoter activity was observed to increase synergistically with introduction of the transcription factor, steroidogenic factor-1 (SF-1). Although SF-1 is a critical mediator of the cAMP response in other genes, mutation of the SF-1 DNA-binding sites in the rat LHbeta gene did not alter the forskolin response nor did forskolin increase SF-1 protein levels in a gonadotrope cell line. In a further set of experiments, it was determined that forskolin-responsiveness was maintained following mutation of the previously defined homeobox-binding element at position -100. We conclude that both Egr-1 and Sp1 contribute to cAMP-dependent transcription of the rat LHbeta gene promoter. While SF-1 does not act independently to mediate the cAMP/PKA response, SF-1 is important for magnification of this response. PMID- 14766010 TI - Estrogen receptor-alpha interaction with the CREB binding protein coactivator is regulated by the cellular environment. AB - The p160 coactivators, steroid receptor coactivator-1 (SRC-1), transcriptional intermediary factor-2 (TIF2) and receptor-associated coactivator-3 (RAC3), as well as the coactivator/integrator CBP, mediate estrogen receptor-alpha (ERalpha) dependent gene expression. Although these coactivators are widely expressed, ERalpha transcriptional activity is cell-type dependent. In this study, we investigated ERalpha interaction with p160 coactivators and CBP in HeLa and HepG2 cell lines. Basal and estradiol (E2)-dependent interactions between the ERalpha ligand-binding domain (LBD) and SRC-1, TIF2 or RAC3 were observed in HeLa and HepG2 cells. The extents of hormone-dependent interactions were similar and interactions between each of the p160 coactivators and the ERalpha LBD were not enhanced by 4-hydroxytamoxifen in either cell type. In contrast to the situation for p160 coactivators, E2-dependent interaction of the ERalpha LBD with CBP or p300 was detected in HeLa but not HepG2 cells by mammalian two-hybrid and coimmunoprecipitation assays, indicating that the cellular environment modulates ERalpha-CBP/p300 interaction. Furthermore, interactions between CBP and p160 coactivators are much more robust in HeLa than HepG2 cells suggesting that poor CBP-p160 interactions are insufficient to support ERalpha-CBP-p160 ternary complexes important for nuclear receptor-CBP interactions. Alterations in p160 coactivators or CBP expression between these two cell types did not account for differences in ERalpha-p160-CBP interactions. Taken together, these data revealed the influence of cellular environment on ERalpha-CBP/p300 interactions, as well as CBP-p160 coactivator binding, and suggest that these differences may contribute to the cell specificity of ERalpha-dependent gene expression. PMID- 14766011 TI - Expression and characterization of recombinant fungal acetyl-CoA carboxylase and isolation of a soraphen-binding domain. AB - Acetyl-CoA carboxylase (ACC) catalyses the first step in fatty-acid biosynthesis. Owing to its role in primary metabolism, ACC has been exploited as a commercial herbicide target and identified as a chemically validated fungicide target. In animals, ACC is also a key regulator of fat metabolism. This function has made ACC a prime target for the development of anti-obesity and anti-Type II diabetes therapeutics. Despite its economic importance, there is a lack of published information on recombinant expression of ACC. We report here the expression of enzymically active fungal (Ustilago maydis ) ACC in Escherichia coli. The recombinant enzyme exhibited Km values of 0.14+/-0.013 mM and 0.19+/-0.041 mM for acetyl-CoA and ATP respectively, which are comparable with those reported for the endogenous enzyme. The polyketide natural product soraphen is a potent inhibitor of the BC (biotin carboxylase) domain of endogenous fungal ACC. Similarly, recombinant ACC activity was inhibited by soraphen with a K(i) of 2.1+/-0.9 nM. A truncated BC domain that included amino acids 2-560 of the full-length protein was also expressed in E. coli. The isolated BC domain was expressed to higher levels, and was more stable than full-length ACC. Although incapable of enzymic turnover, the BC domain exhibited high-affinity soraphen binding (Kd 1.1+/-0.3 nM), demonstrating a native conformation. Additional BC domains from the phytopathogenic fungi Magnaporthe grisea and Phytophthora infestans were also cloned and expressed, and were shown to exhibit high-affinity soraphen binding. Together, these reagents will be useful for structural studies and assay development. PMID- 14766012 TI - Secondary-structure characterization by far-UV CD of highly purified uncoupling protein 1 expressed in yeast. AB - The rat UCP1 (uncoupling protein 1) is a mitochondrial inner-membrane carrier involved in energy dissipation and heat production. We expressed UCP1 carrying a His6 epitope at its C-terminus in Saccharomyces cerevisiae mitochondria. The recombinant-tagged UCP1 was purified by immobilized metal-ion affinity chromatography to homogeneity (>95%). This made it suitable for subsequent biophysical characterization. Fluorescence resonance energy transfer experiments showed that n-dodecyl-beta-D-maltoside-solubilized UCP1-His6 retained its PN (purine nucleotide)-binding capacity. The far-UV CD spectrum of the functional protein clearly indicated the predominance of alpha-helices in the UCP1 secondary structure. The UCP1 secondary structure exhibited an alpha-helical degree of approx. 68%, which is at least 25% higher than the previously reported estimations based on computational predictions. Moreover, the helical content remained unchanged in free and PN-loaded UCP1. A homology model of the first repeat of UCP1, built on the basis of X-ray-solved close parent, the ADP/ATP carrier, strengthened the CD experimental results. Our experimental and computational results indicate that (i) alpha-helices are the major component of UCP1 secondary structure; (ii) PN-binding mechanism does not involve significant secondary-structure rearrangement; and (iii) UCP1 shares similar secondary structure characteristics with the ADP/ATP carrier, at least for the first repeat. PMID- 14766013 TI - Human aldolase A natural mutants: relationship between flexibility of the C terminal region and enzyme function. AB - We have identified a new mutation in the FBP (fructose 1,6-bisphosphate) aldolase A gene in a child with suspected haemolytic anaemia associated with myopathic symptoms at birth and with a subsequent diagnosis of arthrogryposis multiplex congenita and pituitary ectopia. Sequence analysis of the whole gene, also performed on the patient's full-length cDNA, revealed only a Gly346-->Ser substitution in the heterozygous state. We expressed in a bacterial system the new aldolase A Gly346-->Ser mutant, and the Glu206-->Lys mutant identified by others, in a patient with an aldolase A deficit. Analysis of their functional profiles showed that the Gly346Ser mutant had the same Km as the wild-type enzyme, but a 4-fold lower kcat. The Glu206-->Lys mutant had a Km approx. 2-fold higher than that of both the Gly346-->Ser mutant and the wild-type enzyme, and a kcat value 40% less than the wild-type. The Gly346-->Ser and wild-type enzymes had the same Tm (melting temperature), which was approx. 6-7 degrees C higher than that of the Glu206-->Lys enzyme. An extensive molecular graphic analysis of the mutated enzymes, using human and rabbit aldolase A crystallographic structures, suggests that the Glu206-->Lys mutation destabilizes the aldolase A tetramer at the subunit interface, and highlights the fact that the glycine-to serine substitution at position 346 limits the flexibility of the C-terminal region. These results also provide the first evidence that Gly346 is crucial for the correct conformation and function of aldolase A, because it governs the entry/release of the substrates into/from the enzyme cleft, and/or allows important C-terminal residues to approach the active site. PMID- 14766014 TI - Location of promoter elements necessary and sufficient to direct testis-specific expression of the Hst70/Hsp70.2 gene. AB - The rat Hst70 gene and its mouse counterpart Hsp70.2 are expressed specifically in pachytene primary spermatocytes and spermatids. Here we demonstrate that a 165 bp fragment of the Hst70 gene promoter, containing the T1 transcription start site region, entire exon 1 and 42 bp 5' region of the intron, is sufficient to drive testis-specific expression of the chloramphenicol acetyltransferase reporter gene in transgenic mice with the same developmentally regulated pattern as the endogenous Hsp70.2 gene. We show further that high-level tissue-specific gene expression requires additional sequences localized upstream of the T2 transcription start site. Electrophoretic mobility-shift assay analysis revealed that only testes of juvenile rats, when Hst70 gene expression is repressed, contain proteins that specifically bind to the Oct (octamer) sequence localized directly downstream of the T1 site. PMID- 14766015 TI - Concentration-dependent effects of endogenous S-nitrosoglutathione on gene regulation by specificity proteins Sp3 and Sp1. AB - The activities of certain nuclear regulatory proteins are modified by high concentrations of S-nitrosothiols associated with nitrosative stress. In the present study, we have studied the effect of physiological (low microM) concentrations of the endogenous S-nitrosothiol, GSNO (S-nitrosoglutathione), on the activities of nuclear regulatory proteins Sp3 and Sp1 (specificity proteins 3 and 1). Low concentrations of GSNO increased Sp3 binding, as well as Sp3 dependent transcription of the cystic fibrosis transmembrane conductance regulatory gene, cftr. However, higher GSNO levels prevented Sp3 binding, augmented Sp1 binding and prevented both cftr transcription and CFTR (cystic fibrosis transmembrane conductance regulator) expression. We conclude that low concentrations of GSNO favour Sp3 binding to 'housekeeping' genes such as cftr, whereas nitrosative stress-associated GSNO concentrations shut off Sp3-dependent transcription, possibly to redirect cellular resources. Since low micromolar concentrations of GSNO also increase the maturation and activity of a clinically common CFTR mutant, whereas higher concentrations have the opposite effect, these observations may have implications for dosing of S-nitrosylating agents used in cystic fibrosis clinical trials. PMID- 14766016 TI - Inosine 5'-monophosphate dehydrogenase binds nucleic acids in vitro and in vivo. AB - Inosine 5'-monophosphate dehydrogenase (IMPDH) is the rate-limiting enzyme in the de novo biosynthesis of guanine nucleotides. In addition to the catalytic domain, IMPDH contains a subdomain of unknown function composed of two cystathione beta synthase domains. Our results, using three different assays, show that IMPDHs from Tritrichomonas foetus, Escherichia coli, and both human isoforms bind single stranded nucleic acids with nanomolar affinity via the subdomain. Approx. 100 nucleotides are bound per IMPDH tetramer. Deletion of the subdomain decreases affinity 10-fold and decreases site size to 60 nucleotides, whereas substitution of conserved Arg/Lys residues in the subdomain with Glu decreases affinity by 20 fold. IMPDH is found in the nucleus of human cells, as might be expected for a nucleic-acid-binding protein. Lastly, immunoprecipitation experiments show that IMPDH binds both RNA and DNA in vivo. These experiments indicate that IMPDH has a previously unappreciated role in replication, transcription or translation that is mediated by the subdomain. PMID- 14766019 TI - Use of aminophylline in the treatment of severe symptomatic bradycardia resistant to atropine. AB - Bradycardia and cardiac arrest are known complications of acute spinal cord injuries and are usually temporary. If the general measures of correcting hypoxia and using atropine fail, placement of a temporary followed by a permanent pacemaker is typically considered. We describe 2 very interesting cases of severe symptomatic bradycardia resistant to atropine, where we were able to obviate the use of pacemaker placement by the simple use of intravenous aminophylline. Aminophylline had been used in the past for treating resistant bradycardia in settings such as acute inferior wall myocardial infarction, cardiac transplantation, and so on, but has never been used in the setting of acute spinal cord injuries. Aminophylline probably works in this setting by increasing cyclic adenosine monophosphate (cAMP) and activating the sympathoadrenal system. PMID- 14766018 TI - Identification of a POU factor involved in regulating the neuron-specific expression of the gene encoding diapause hormone and pheromone biosynthesis activating neuropeptide in Bombyx mori. AB - Diapause hormone (DH) and PBAN (pheromone biosynthesis-activating neuropeptide) are two important insect neuropeptides regulating development and reproduction respectively. In the present study, we report two Bombyx mori transcription factors interacting specifically with the promoter of Bom-DH-PBAN (where Bom-DH stands for B. mori DH); we named them DHMBP-1 and -2 (DH-modulator-binding proteins 1 and 2). The developmental changes of DHMBP-1/-2 are closely correlated with that of Bom-DH-PBAN mRNA throughout the pupal stage. Competition assays indicate that DHMBP-1 from Chinese B. mori possesses binding characteristics similar to those of the POU-M1 protein from Japanese B. mori. POU-M1 cDNAs were cloned from various tissues of Chinese B. mori and were found to be distinct from the previously published POU-M1 in amino acid residues 108-136 because of insertion mutations. Owing to this difference in amino acid residues, we named this cDNA POU-M2. Even though POU-M2 differs from POU-M1 at the N-terminal, the POU domain and the binding properties of both POU-M1 and -M2 are the same. Functional analysis showed that overexpression of POU-M2 in the Bombyx cell line BmN activated the promoter of Bom-DH-PBAN, but failed to activate a promoter in which the POU-binding element was mutated. The transcriptional activity of POU-M2 is probably regulated by other factors binding to the upstream of the promoter sequence. We show that the POU-M2-binding site was able to activate the transcription of a heterologous promoter of the gene encoding B. mori larval serum protein. POU-M1 was found to exhibit the same transcriptional activities as POU-M2. Taken together, these results demonstrate that POU-M2 plays an important role in the transcriptional regulation of the Bom-DH-PBAN gene. PMID- 14766017 TI - Cell-cycle-dependent translation of histone mRNAs is the key control point for regulation of histone biosynthesis in Leishmania infantum. AB - The cell-cycle-dependent expression of the four core histones (H2A, H2B, H3 and H4) has been studied in the protozoan parasite Leishmania infantum. For that purpose, the cell cycle was arrested by incubation of promastigotes with the DNA synthesis inhibitor hydroxyurea, which induced an accumulation of cells stalled in G1 phase. Hydroxyurea release resulted in a semi-synchronous entry into the cell cycle, as determined by flow cytometry. The steady-state levels of histone mRNAs in the G1, S and G2/M phases were found to be constant along the cell cycle. However, the levels of histone synthesis increased when parasites enter the S phase, in agreement with previous results showing that histone synthesis in Leishmania is tightly coupled with DNA replication. In addition, we analysed the distribution of histone mRNAs on polyribosomes at different stages of the cell cycle by separation of cytoplasmic RNAs in sucrose gradients. Remarkably, a drastic change in the polysome profiles of histone mRNAs was observed during the progression from G1 to S phase. Thus, in the S phase, histone mRNAs are present in ribosome-bound fractions, but in the G1 phase, the histone transcripts are exclusively found in the ribosome-free fractions. These results support a regulatory model in which the cell-cycle-regulated synthesis of histones in Leishmania is controlled through a reversible interaction between translational repressors and histone mRNAs. PMID- 14766020 TI - A 38-year-old woman with dizziness. PMID- 14766021 TI - Cardiac chagas disease masquerading as an acute myocardial infarction. AB - Although myocardial infarction is most often the manifestation of epicardial coronary artery disease, Chagas heart disease due to chronic Trypanasome Cruzi infection may present with a syndrome of chest pain and elevations in markers of cardiac myonecrosis. In the setting of an increasingly diverse global population and immigration of peoples from endemic areas of Trypanasome Cruzi, it is important to be aware of the myriad cardiac manifestations of Chagas disease. PMID- 14766022 TI - Vitamins, supplements, herbal medicines, and arrhythmias. AB - Nutritional and herbal supplements may have harmful or beneficial effects on arrhythmias. Potential supplements that may have antiarrhythmic activity include omega-3 polyunsaturated fatty acids (N-3 PUFA), coenzyme Q10, and carnitine. Clinical studies show that N-3 PUFA or fish oil supplementation appears to reduce mortality and sudden death. Coenzyme Q10, used in treatment of heart failure, and carnitine and its derivatives may have beneficial effects on arrhythmias, although clinical studies have been limited. Antioxidant supplements may be beneficial, but large studies with vitamin E have been disappointing in that it does not reduce mortality. Correction of electrolyte disturbances has been long advised and magnesium supplementation has been beneficial in the treatment of torsades de pointes and in some studies after cardiac surgery. However, routine electrolyte supplementation with empiric potassium or magnesium in non-deficient patients has not been convincingly beneficial. Several herbal supplements have also been promoted to have antiarrhythmic activity. However, clinical studies are lacking to support routine use of these herbal medications. In addition, some herbal supplements may cause serious proarrhythmia, and many supplements significantly interact with warfarin and digoxin. PMID- 14766023 TI - Magnetism and cardiac arrhythmias. AB - Low-level electromagnetic fields (EMFs) have been used to treat various neurologic disorders. In the present study, we applied micro Gauss (microG) levels of EMFs either to the vagosympathetic nerve trunks, dissected in the neck, or across the chest in anesthetized dogs. Based on theoretical and empiric grounds, we compared EMFs (2.87 microG at 0.043 Hz) delivered to the vagosympathetic trunks in an experimental set (n = 5) with a sham control group (n = 6). Over a period of 2 to 3 hours, heart rate decreased after an initial 5 minute EMF exposure. The maximal heart rate changes in the experimental versus control groups was 29% versus 12% (P = 0.03). The voltage applied to the autonomic nerves required to induce atrioventricular (AV) conduction block decreased by 60% in the experimental group versus a 5% increase in the control group (P = 0.005). This effect also lasted 2 to 3 hours. Another EMF setting (amplitude 0.34 microG, frequency 2 kHz) applied for 5 minutes to the vagosympathetic trunks was associated with a significant increase in the occurrence of atrial premature depolarizations (APDs), atrial tachycardia (AT), and atrial fibrillation (AF) in response to autonomic nerve stimulation compared with control states before EMF exposure. No atrial arrhythmias could be induced after propranolol and atropine, even at the highest voltage used to stimulate the autonomic nervous input to the heart (n = 11). Only 2 dogs showed no response to this EMF application. In 3 dogs in whom atrial pacing (cycle length = 250 ms) and autonomic nerve stimulation induced AF, an EMF (2.87 microG at 0.043 Hz) delivered for 35 minutes across the chest suppressed AF for up to 3 to 4 hours, after which the same protocol again induced AF. We conclude that in these preliminary experiments, specific low-level EMFs alter heart rate, AV conduction, and heart rhythm. These effects were mediated through the autonomic nervous system inputs to the heart based on adjunctive effect of autonomic nerve stimulation and the inhibitory action of autonomic blockers. PMID- 14766024 TI - The influence of gender on arrhythmias. AB - It is becoming increasingly apparent that there are important gender differences in normal cardiac physiology. These, in turn, could be associated with differences in the type and severity of cardiac arrhythmias. Women have higher resting heart rates than men, probably as a result of a combination of autonomic and intrinsic factors. The clinical significance of this observation is unclear at the present time. Women have a lower incidence of sudden cardiac death, which could be related to the delayed onset of coronary artery disease in women, presumably as a result of the protective effects of female hormones during gestational years. In survivors of sudden cardiac death, there are major gender differences, with fewer women having underlying coronary artery disease and a greater percentage of women having structurally normal hearts. QT interval prolongation and Torsade de Pointes are more common in women, probably on the basis of differences in ion channels between genders. Women appear especially susceptible to Torsades from QT-prolonging drugs such as quinidine or tricyclic antidepressant medications. The mechanisms of paroxysmal supraventricular tachycardia differ between the genders, although therapy seems to be equally efficacious in men and women. Lastly, atrial fibrillation is considerably more common in men. There is also some evidence that it is better tolerated by men. PMID- 14766025 TI - Giant ascending aortic aneurysm with type a dissection diagnosed by aortography. PMID- 14766026 TI - High-density lipoprotein cholesterol and coronary heart disease. AB - There is a large body of evidence demonstrating an inverse correlation between circulating levels of high-density lipoprotein (HDL) cholesterol and cardiovascular disease risk. For every 1-mg/dL increase in HDL, it is estimated that the risk of cardiovascular events decreases by 2% to 3%. HDL is one of many factors that contribute to the regulation of the atherosclerotic process. HDL mediates reverse cholesterol transport and exhibits numerous beneficial properties, including antioxidant, antiinflammatory, and antithrombotic effects on the vasculature. Recent studies have expanded our understanding of the vasoprotective mechanisms of HDL to include enhanced nitric oxide production and improved endothelium-dependent relaxation. Progress has also been made in determining the molecular mechanisms that mediate reverse cholesterol transport. Recently published National Cholesterol Education Program Adult Treatment Panel guidelines have broadened the definition of low levels of HDL and encourage more aggressive screening and treatment of lipid abnormalities. Several therapeutic interventions can augment HDL concentrations, and there is increasing evidence that these interventions improve cardiovascular outcomes. Research focusing on defining the molecular roles of HDL will likely identify potential therapeutic targets for decreasing the incidence and progression of coronary heart disease. This review highlights the role of HDL in coronary heart disease, from basic mechanisms of action to recent clinical trial results. PMID- 14766027 TI - Emerging role of integrative medicine in cardiovascular disease. AB - Our evolution toward a more integrative approach toward healing will accelerate if we can alter the different perspectives that patients and physicians bring to their relationship. This article reviews lessons that have been acquired using alternative approaches to facilitate the recovery of patients undergoing invasive procedures. After identifying that most of our patients use alternative therapies but prefer not to discuss these therapies with their surgeons, we began to routinely refer our patients to a coordinator trained in this field. The resulting integrative medicine program offers massage, yoga, audiotapes, and additional customized treatments. The coexisting research laboratory designs and conducts well-organized research endeavors evaluating the efficacy of therapies being offered. By mixing clinical practice with conventional research, we seek to emulate the successful formula used in allopathic medicine. PMID- 14766028 TI - A standard method for measuring blood pressure in the Americas. PMID- 14766029 TI - Rubella and the Americas. PMID- 14766030 TI - Working meeting on blood pressure measurement: suggestions for measuring blood pressure to use in populations surveys. AB - As part of the Pan American Hypertension Initiative (PAHI), the Pan American Health Organization and the National Heart, Lung, and Blood Institute of the National Institutes of Health of the United States of America conducted a working meeting to discuss blood pressure (BP) measurement methods used in various hypertension prevalence surveys and clinical trials, with the objective of developing a BP measurement protocol for use in hypertension prevalence surveys in the Americas. No such common protocol has existed in the Americas, so it has been difficult to compare hypertension prevention and intervention strategies. This piece describes a proposed standard method for measuring blood pressure for use in population surveys in the Region of the Americas. The piece covers: considerations for developing a common blood pressure measurement protocol, critical issues in measuring blood pressure in national surveys, minimum procedures for blood pressure measurement during surveillance, and quality assessment of blood pressure. PMID- 14766031 TI - [Reference values for erythrocyte cholinesterase activity in the working population of Antioquia, Colombia, according to the Michel and EQM techniques]. AB - OBJECTIVE: To establish reference values for erythrocyte cholinesterase (EC 3.1.1.7) activity for the active working population of two regions of the department of Antioquia, Colombia, that are located at different altitudes above sea level. METHODS: We took representative samples from populations of active working persons 18 to 59 years old from two regions in the department of Antioquia: (1) the Aburra Valley (1 540 m above sea level) and (2) the near east of the department (2 150 m above sea level). We excluded workers who were using cholinesterase-inhibiting substances in their work or at home, those who had a disease that altered their cholinesterase levels, and those who said they were not in good health. We measured the erythrocyte cholinesterase activity using two methods: (1) the Michel method and (2) the EQM method (EQM Research, Cincinnati, Ohio, United States of America). We carried out the measurements with 827 people, 415 from the Aburra Valley and 412 from the near east region. We compared proportions using the chi-square test and Fisher's exact test. We utilized the Student's t test for independent samples to compare two averages. To simultaneously compare three or more averages, analysis of variance was used, followed by the Newman-Keuls multiple-range test. When the variables were not normally distributed or when the variances were not homogeneous, Kruskal-Wallis nonparametric analysis of variance was used to compare the medians. Three computer software programs were used in the statistical analysis: SPSS 9.0, SGPlus 7.1, and Epi Info 6.04. In all the statistical tests the level of significance was set at P < 0.05. RESULTS: The average erythrocyte cholinesterase activity value that we found for the studied population by using the Michel method was 0.857 delta pH/hour (95% confidence interval (CI): 0.849 to 0.866), and the average value found through the EQM method was 35.21 U/g hemoglobin (95% CI: 34.82 to 35.60). With the Michel method: (1) the enzymatic activity differed significantly between the two regions, according to the Newman-Keuls test; (2) within each region, the enzymatic activity was significantly higher among males than among females, according to the Newman-Keuls test; and (3) in none of the region-sex strata was there a statistically significant influence of age on the enzymatic activity. Using the EQM method, there were no statistically significant differences by region, sex, or age group. CONCLUSIONS: The erythrocyte cholinesterase activity values found by the two analytical techniques were significantly higher than the values from outside Colombia that are now being used as reference values in the country, which poses both clinical and epidemiological problems. We recommend that the data from this study be adopted as the reference values in Colombia. PMID- 14766032 TI - [Indicators of the effectiveness of epidemiological surveillance for acute flaccid paralysis in Brazil from 1990 through 2000]. AB - OBJECTIVES: To describe and compare indicators of the quality of epidemiological surveillance for acute flaccid paralysis (AFP) before Brazil's certification as a poliomyelitis-free area (1990 through 1994) and after that certification (1995 through 2000). METHODS: The following variables were studied: minimum prevalence (reporting annually at least 1 case of AFP per 100 000 individuals younger than 15 years), negative reporting (weekly reporting of the absence of AFP cases by at least 80% of the reporting units in each region), time of investigation (investigating at least 80% of the AFP cases within 48 hours of their being reported), and diagnostic investigation (collection of two stool samples for viral culture in the 2 weeks following the onset of paralysis in 80% or more of the AFP cases). The results were categorized as adequate if they were equal to or higher than the value considered as being effective, or as inadequate if they were below this value. The percentage rate of adequate values was calculated for each of Brazil's 27 federal political divisions (26 states plus the Federal District) in each year, for each of the two periods studied. The final means per period, per federal political division, and per region were also calculated. RESULTS: For Brazil overall, minimum prevalence decreased from 1.3 per 100 000 individuals younger than 15 years in the precertification period to 0.9 per 100 000 in the postcertification period, but the difference was not statistically significant (P > 0.08). While negative reporting was adequate in both periods (87% and 84%, respectively), the percentage rate of effectiveness was 6.6% lower in the postcertification period (P > 0.21). The effectiveness in the time required to investigate AFP cases increased from 60.7% to 71.6% (P > 0.06). The collection of stool samples was inadequate both before and after certification (54% and 52%, respectively). CONCLUSIONS: Despite the absence of indigenous wild virus poliomyelitis cases in Brazil in the postcertification period, the quality of epidemiological surveillance indicators is still less than desirable. This may delay the detection and investigation of AFP cases and compromise the global poliomyelitis eradication effort. New information strategies that are being made available for the health system in Brazil may improve the effectiveness of the national poliomyelitis control program. PMID- 14766033 TI - Serological surveillance of measles in blood donors in Rio de Janeiro, Brazil. AB - OBJECTIVES: To estimate the proportion of individuals seronegative for measles antibody among blood donors from a blood bank in the city of Rio de Janeiro, Brazil, and to describe their social and demographic characteristics, with the goal of exploring the potential use of plasma banks to supplement serological surveillance with relevant data in order to support the program of measles elimination in the city of Rio de Janeiro and elsewhere. METHODS: Plasma samples from 1 101 consecutive blood donations made in November 2000 at HEMORIO, the largest blood bank in the state of Rio de Janeiro, were tested for measles immunoglobulin G, using a commercial enzyme immunoassay and a plaque reduction neutralization test. We calculated the proportion (and 95% confidence interval (CI)) of samples seronegative for measles antibody for the total sample of blood donors and also for subgroups categorized by age, sex, neighborhood of residence, education, and occupation. We used the chi-square test to assess the statistical significance of differences in proportions and linear trends in proportions. RESULTS: Of the total group of blood donors, 6.9% of them (95% CI: 5.4%-8.4%) were seronegative for measles. Women had a higher proportion (10.1%; 95% CI: 6.8% 13.4%) of seronegative results than did men (5.6%; 95% CI: 4.0%-7.2%). In terms of age, 86.8% of seronegative individuals were born between 1971 and 1982. Seronegativity was inversely proportional to age (chi-square = 58.0; P < 0.0001). With regard to occupation, students had the highest proportion of seronegative individuals (17.8%). In terms of education, most of the susceptible persons were in the categories of "incomplete university degree" or "incomplete high school." For the various areas of residence the proportions ranged from 2.1% to 11.4%. CONCLUSIONS: Blood bank plasma may constitute a useful and convenient source of complementary data for serological surveillance in adults of measles and other infections for which immunization and surveillance activities are implemented. This approach could be beneficial to other areas in Brazil and other countries where plasma from blood donors is available for surveillance. The use of residual sera from patients and plasma from blood donors represents a tradeoff between representativeness and timeliness as well as economy of resources. PMID- 14766034 TI - [Fluoride in drinking water in Cuba and its association with geological and geographical variables]. AB - OBJECTIVE: To determine the association between different concentrations of the fluoride ion in drinking water and some geological and geographical variables in Cuba, by using a geographic information system. METHODS: From November 1998 to October 1999 we studied the fluoride concentration in the sources of drinking water for 753 Cuban localities that had at least 1 000 inhabitants. For the information analysis we utilized the MapInfo Professional version 5.5 geographic information system, using the overlaying method. The study variables were the concentration of the fluoride ion in the water sources, the geological characteristics of the area, the alignments (geological characteristics that were found together), the types of water sources, and whether an area was a plain or mountainous. The results were grouped by locality and municipality. RESULTS: In 83.1% of the localities, the water samples were collected from wells and springs, and the remaining 16.9% came from dams and rivers. Of the 753 localities studied, 675 of them (89.6%) had low or medium fluoride concentrations (under 0.7 mg/L). The eastern region of the country was the one most affected by high fluoride concentrations in the waters, followed by the central region of the country. The majority of the localities with high natural fluoride concentrations were in areas located on Cretaceous volcanic arc rocks. The presence of fluoride in the drinking waters was related to the alignments with the earth's crust, in rock complexes of volcanic-sedimentary origin and of intrusive origin and also in carbonate rocks. However, the highest fluoride concentrations generally coincided with rock complexes of volcanic-sedimentary origin and of intrusive origin. All the localities with high fluoride concentrations in the water were associated with wells. CONCLUSIONS: The fluoride concentration is low or medium in the drinking water sources for 89.6% of the Cuban localities with at least 1 000 inhabitants. Geological and geographical characteristics can help identify areas with optimal or high concentrations of the fluoride ion in the drinking water. PMID- 14766035 TI - [Missed opportunities for tetanus immunization of pregnant women in Juiz de Fora, Minas Gerais, Brazil]. AB - OBJECTIVE: To estimate the missed opportunities for tetanus vaccination and to determine the immunization coverage at the end of pregnancy in women living in the city of Juiz de Fora, which is in the state of Minas Gerais, Brazil. METHODS: A cross-sectional study was carried out of 430 women who were randomly selected from all the maternity hospitals in the city, from February through August 1996. The women were interviewed regarding prenatal care, their knowledge of tetanus, and tetanus immunization before or during the latest pregnancy. RESULTS: The age of the participants ranged from 14 to 45 years (mean = 26.4 +/- 6.9 years); 420 women lived in urban areas and 10 in rural areas. Prenatal care was from the Brazilian public Unified Health System in 69.5% of the women (299 of 430); 27.6% (119) received prenatal care through other types of providers; and 2.8% (12) did not receive prenatal care. Of the 430 participants, 352 of them (81.8%) had their prenatal care card with them during the interview; 85.6% (368) had four or more prenatal care visits. Regarding their knowledge of tetanus, 92.1% (396) knew about the disease and its severity. Out of the 430 women, 359 of them (83.5%) started the latest pregnancy without having had a tetanus vaccination; 104 were vaccinated during the latest pregnancy, so 255 had not been vaccinated by the time of delivery. This was in spite of the significant increase in the opportunities for being vaccinated with an increase in the number of prenatal visits. The odds ratio for pregnant women with four or more prenatal visits being vaccinated by the end of the pregnancy was 2.7 times that of the women with zero to three visits. Seventy percent of the opportunities for tetanus immunization were missed in the study population. The overall tetanus immunization coverage was 40.6% by the end of pregnancy. CONCLUSIONS: The low immunization coverage was below the level recommended by the World Health Organization. In combination with the high rate of missed opportunities for vaccination, this points to the need to implement a tetanus vaccination promotion strategy directed at health professionals in general but especially at those who provide care for pregnant women, in both public and private health clinics. In addition, the vaccination practices in Juiz de Fora should be strengthened. PMID- 14766037 TI - [New goal for vaccination programs in the Region of the Americas: to eliminate rubella and congenital rubella syndrome. 2003]. AB - The vaccination programs of the countries in the Region of the Americas have produced notable results over the last several decades. In recognition of those accomplishments, in June 2003 the 132nd session of the Executive Committee of the Pan American Health Organization (PAHO) called on the Member States of PAHO to prepare, within one year, national action plans for the elimination of rubella and congenital rubella syndrome (CRS) by the year 2010. During the Executive Committee discussions, praise was given for the sustained efforts of the countries of the Region in the struggle to eradicate measles. The Executive Committee discussions also reiterated the recommendation that the countries establish specific line items for immunization within their national budgets, in order to protect the investments in immunization that the Region's countries have made. The analysis of results of mass vaccination against rubella indicates that the benefits coming from accelerated control greatly exceed the costs associated with the treatment and rehabilitation of children with CRS. Various factors have created an environment that supports the goal of eliminating rubella and CRS from the Americas by the year 2010. These factors include the rapid reduction in morbidity that has come from the accelerated rubella control strategy; the availability of a safe, affordable, efficacious vaccine; the evidence of the cost benefit of vaccinating against rubella; and the broad support provided by the public and by health authorities in the Member States of PAHO. PMID- 14766038 TI - Promoting the healthy, safe use of recreational waters. 2003. PMID- 14766039 TI - [Treatment of portal hypertension by endoscopic esophageal ligation of varicose veins combined with partial splenic embolization]. AB - OBJECTIVE: To evaluate the feasibility and efficacy of a new method of endoscopic esophageal variceal ligation combined with partial splenic embolization (EVL-PSE) for the patients with portal hypertension. METHODS: From May 1999 to February 2003, sixty-eight patients with portal hypertension underwent EVL-PSE, and hemodynamics of the portal trunk (PT), the left gastric vein and azygos vein, including maximum velocity, flow volume, vein diameter, were assessed using color ultrasound Doppler. RESULTS: The esophageal varices and hypersplenism were greatly ameliorated after operation in patients who had undergone EVL-PSE. Postoperative portal trunk flow volume and velocity were significantly reduced (P < 0.05), and flow volume of the left gastric vein as well as the azygos vein were also reduced after operation. During 2 - 24-month follow-up, no recurrent bleeding was found. CONCLUSIONS: EVL-PSE is less traumatic with less complications, and results in marked eradication of esophageal varices, it can be carried out safely in the clinical treatment for patients with portal hypertension. PMID- 14766040 TI - [Prevention and treatment of complications of laparoscopic cholecystectomy]. AB - OBJECTIVE: To investigate the prevention and the treatment of the complications of laparoscopic cholecystectomy, including bile duct injury, bile leakage and bleeding. METHODS: 1,100 patients undergoing laparoscopic cholecystectomy from October 1993 to January 2003 were retrospectively analyzed. RESULTS: Operative complications included bile duct injury in one case (0.09%), bile leakage in three cases (0.27%), and bleeding in four cases (0.36%). All these complications were cured using operation and non-operation methods. CONCLUSIONS: To prevent bile duct injury, bile leakage as well as bleeding, abnormal cystic duct and cystic artery should be identified, and electric damage to the surrounding tissues should be avoided during operation. PMID- 14766041 TI - [Candida infection in patients with acute necrotizing pancreatitis]. AB - OBJECTIVE: To summarize our hospital's experience in the diagnosis and treatment of Candida infection in patients with acute necrotizing pancreatitis (ANP). METHODS: Seventy-eight cases with ANP were reviewed. There were diagnoses either by operative finding or by CT scanning. Sixty-two cases received prophylactic antibiotic treatment, other sixteen did not. For cultivation of Candida, blood, urine, stool, sputum and wound drainage fluid culture, and swabs were examined microbiologically for fungi. RESULTS: The incidence of Candida infection in all patients with ANP was 17.9% (14/78) and mortality was 28.6% (4/14). The incidence of prophylactic antibiotic group was 19.4% (12/62) and mortality was 25.0% (3/12). Non prophylactic group was 12.5% (2/16) and 50.0%. CONCLUSIONS: Our data provide evidence for the clinical significance of Candida infection in patients with ANP. The current prophylactic antibiotic treatment can prevent a septic course of the ANP, but might lead to the evolution of Candida infection. PMID- 14766042 TI - [Long-term therapeutic effects of total gastrectomy in cancer of the cardia and stomach fundus]. AB - OBJECTIVE: To evaluate total gastrectomy for the treatment of cancer of the cardia and stomach fundus. METHODS: Five hundred and thirteen patients with cancer of the cardia and stomach fundus underwent radical resection. Of them, 326 were treated using total gastrectomy (group TG); and 187, using proximal gastrectomy (group PG). The 5-year and 10-year survival rates and the postoperative complication rate and mortality rate were followed up and compared in the two groups. RESULTS: The 5-year and 10-year survival rates of group TG were 43.6% and 24.5%, of group PG were 33.9% and 14.1%, respectively, and the difference was statistically significant (chi(2) = 4.421, P < 0.05, chi(2) = 5.726, P < 0.05). The postoperative complication rate and mortality rate of group TG were 14.7% and 3.1%, of group PG were 10.2% and 2.1%, respectively, and the difference was not statistically significant (chi(2) = 1.796, P > 0.05, chi(2) = 0.082, P > 0.05). CONCLUSIONS: To improve long-term therapeutic effects, total gastrectomy should be recommended for stage III patients with cancer of the cardia and stomach fundus when tumor size is bigger than 3.0 cm or lymph node metastasis occur. The postoperative complication rate and mortality rate should not be increased and the esophagitis of gastroesophageal reflux should be avoided in the patients treated using total gastrectomy. PMID- 14766043 TI - [Using ultrasonically activated scalpels as major instrument for vessel dividing and bleeding control in minimally invasive video-assisted thyroidectomy]. AB - OBJECTIVE: To establish new techniques on the control of vessel dividing and bleeding in minimally invasive video-assisted thyroid surgeries. METHODS: One hundred and seventy volunteer patients with nodular goiter, adenoma and Graves' disease consecutively received the endoscopic thyroid operation according to Miccoli's mode from April 2002 to June 2003. The approach was designed to use ultrasonically activated scalpels [Harmonic Scalpel (R), Johnson & Johnson, USA] as a major, suction-dissector or others as supplementary instruments during the whole process. During the operations, two methods, "sequenced dissect-coagulate cut" and "pre-coagulation + interlaced cut-suck-coagulate-dissect", were employed and separately evaluated for their safety or efficacy. RESULTS: All of the procedures, except that 2 (their lesions were proven to be "cancer" by frozen section and need different treatment), were successfully completed. None of them were interrupted and converted to open surgery due to uncontrolled bleeding or severe postoperative hematoma. Branches of major thyroid vessels can be directly divided by ultrasonic scalpel without ligation or using hemoclips, supposing the technique of "sequenced dissect-coagulate-cut" method was strictly used. Bleeding during intra-gland dissection can be effectively controlled by the combined techniques of precoagulation of the vessels and the capsule vessel network near and around the incision, and then, by coordinating manipulation with a special multiple functional ultrasonic scalpel and a suction-dissector, supposing the technique of "interlaced cut-suck-coagulate-dissect" was properly employed. CONCLUSIONS: Using ultrasonic scalpels as major, suction-dissector or others as supplementary instruments, manipulating cooperatively during the whole process, can be an ideal fashion in the site of a narrow gasless working space via a small single incision in anterio-inferior neck. And this may dramatically simplify the manipulation, and hence greatly reduce the operative difficulty. The reforms resolved two key technique problems in Miccoli's endoscopic operation: vessel dividing and bleeding-control, therefore, laying a sound basis for the clinical application of this special approach. PMID- 14766044 TI - [Biological behavior of cardial cancer and its effect on surgical management]. AB - OBJECTIVE: To study the biological behavior of cardial cancer and its influence on surgical management. METHODS: Complete clinicopathologic data of 46 cases with cardial cancer undergoing radical gastrectomy was investigated retrospectively. The relationships between tumor Borrmann type, depth of invasion, growth pattern, lymph node metastasis and 5-year survival rate postoperative were analyzed. RESULTS: Of 46 cases, Borrmann type III, Type IV and Type II was 76% (35/46), 18% (8/46) and 6% (3/46) respectively; 5-year survival rate was 40% (14/35), 0 (0/8) and 100% (3/3) respectively. In respect of the depth of invasion, pT(2) was 31% (14/46) cases with 71% (10/14) lymph node metastasis; and 5-year survival rate was 64% (9/14). pT(3) was 15% (7/46) cases with 86% (6/7) lymph node metastasis; and 5-year survival rate was 57% (4/7). And pT(4) was 54% (25/46) cases with 92% (23/25) lymph node metastasis; and 5-year survival rate was 12% (3/25). The growth pattern in 87% (40/46) cases was infiltrative; and 5-year survival rate was 28% (11/40); the growth pattern in 13% (6/46) cases was expansive; and 5-year survival rate was 100% (6/6). CONCLUSION: D(2)(+) radical total gastrectomy should be performed on the developed cardial cancer, and if necessary, resection of body and tail of pancreas should be chosen. PMID- 14766045 TI - [Routine use of Brener shunt during carotid endarterectomy]. AB - OBJECTIVE: To evaluate the routine use of Brener shunt during carotid endarterectomy (CEA). METHODS: A prospective audit of the results of 58 consecutive CEA was performed, with routine use of Brener shunt in surgical hospital, Rostock university in Germany. Intraoperative and postoperative complications of CEA were reviewed. RESULTS: In this study, the successful rate of insertion of shunt was 100%. The perioperative stroke/death rate at 30 days was 0, but the TIA rate was 2 cases (3.5%), which was related with previous stroke and contralateral internal carotid occlusion. 2 cases (3.5%) had minor operative complications. CONCLUSIONS: The routine use of Brener shunt has a less rate of intraoperative and early postoperative neurologic complications during CEA. But experience in a large number of shunting procedure is needed. PMID- 14766046 TI - [The reconstruction of three-dimension morphology of proximal femur]. AB - OBJECTIVES: To reconstruct the morphology of the proximal femur and measure the parameters of them by means of the computer-aided design (CAD) in order to provide help to the design and research of the domestic prosthesis, event for the selection of the adapted implant for the doctors before operation. METHODS: The data of 160 healthy Chinese proximal femur images were obtained by the PHLLIP SR7000 computer tomography. The correlative parameters were measured exactly by medical imaging software. Then the imaging data were input into an computerized contouring program which generates the three dimensional coordinates representing periosteal and endosteal contours of the femur for each CT slice. Though the computer-aided design (CAD) software, the three-dimensional solid model was reconstructed by the vectorisation of image contours. In the circumstance of CAD system, the bone axis is defined as the line across the geometric center of the cross-section of the isthmus and 20 mm above the lesser trochanter mid-point. We measured the parameters of the endosteal and periosteal dimensions of the medullary canal. RESULTS: The parameters of the medullary canal measured with the reconstructed normal Chinese femur have significant difference from that measured with the Caucasian's. The extracortical and endosteal parameters of male have significant differences from most of female's. However, there were no significant differences in endosteal width between different age groups in two gender groups. From the proximal to the distal femur, the intersection angles between the longest mediolateral endosteal diameters of the slices and the plane of the femoral neck were increased. CONCLUSIONS: The reconstructive femur contains more information and can be cut at arbitrary levels which means it is facility to the research of the Morphology of the Proximal femur and the measurement of relative parameters. It is necessary to design the prosthesis which fit anatomic parameters of the Chinese femur. It needs to further study whether the gender factor will be considered in the prosthesis design. The angle between the anteversion and the helitorsion at the level 20 mm above the lesser trochanter midpoint should be noticed by the implant designers and doctors. PMID- 14766047 TI - [Clinical study of solitary bone plasmacytoma]. AB - OBJECTIVE: To study the clinical characteristics and the prognosis of solitary bone plasmacytoma (SBP). METHODS: From June 1997 to June 2002, ten SBP patients were treated in our department. Two lesions were in sacrum, two in pelvis, the other six lesions were in proximal femur, femoral diaphyseal, scapula, thoracic vertebra, proximal humus, and rib respectively. The preoperative laboratory tests were normal. Operation was performed on all patients. The pathologic diagnose was plasmacytoma and then they underwent further laboratory examination, urine Bence Jone protein is positive in 3 patients, serous IgG value was higher than normal in 2 patients, abnormal M protein was found in electrophoresis in 2 patients. All patients received radiotherapy postoperatively. RESULTS: Four patients were excluded, whose follow-up were lessen than ten months, the other six patients's average follow-up is 28.2 months (from 18 to 48 months), one patient who developed multiple myeloma (MM) six months postoperatively received chemotherapy using M2 protocol and died 21 months after operation. The other five patients had disease-free survive and remain solitary bone lesion after the treatment of surgery and radiology. CONCLUSION: Compared with MM, SBP patients are younger, the therapeutic results and prognosis are better. The main prognostic factors include age, the size of lesion, the axial bone lesion, persistence of myeloma protein after radiotherapy, early diagnosis and treatment, and so on. PMID- 14766048 TI - [Diagnosis and treatment for pulmonary thromboembolism in postthoracotomy patients]. AB - OBJECTIVE: To report the experience in the diagnosis and treatment of pulmonary thromboembolism (PTE) in postthoracotomy patients and to analyze current problems in this field. METHODS: From January 2001 to June 2002 we diagnosed and treated 5 patient who had pulmonary thromboembolism, 72-168 hours after thoracotomy (male 4, female 1, mean age 63 years). Symptoms include breathholding, chest pain, palpitation, coma and so on. Physical signs include tachypnea, hypotension and tachycardia. We utilized spiral CT pulmonary artery angiography (SCTPA) and Doppler echocardiography to judge PTE. Pulmonary artery interventional therapy was used in 3 cases and total body thrombolysis in 1. RESULTS: 4 patients recovered with no complication and recurrence; 1 patient died from congestive heart failure and respiratory failure. CONCLUSION: Deep venous thrombosis (DVT) and PTE are related to the surgical intervention, chest tumor and thoracotomy is high risk factors. Screenage examination is primary to used. Pulmonary artery interventional therapy is safe to the postthoracotomy patients. Preventive measures is very important in the high-risk group. PMID- 14766049 TI - [Radionuclide pulmonary perfusion imaging in evaluating pulmonary hypertension in patients with rheumatic heart disease]. AB - OBJECTIVE: To evaluate the degree of pulmonary hypertension in patients with rheumatic heart disease using radionuclide pulmonary perfusion imaging. METHODS: The pulmonary perfusion in 25 patients with rheumatic heart disease was examined using scintigram with macroaggregates of (99m)Tc-labeled human serum albumin (PPS) before and 7 days after operation. PPS was analyzed for (1) pulmonary perfusion steady time (PT), right upper and lower lung count ratio (RULR). The results were compared with those of catheterization examination during the operation. RESULTS: The pulmonary arterial systolic pressure (PAs) and total pulmonary resistance (TPR) were (60 +/- 21) mm Hg and (421 +/- 106) dyn if PT > or = 20 seconds and RULR > or = 2; The PAs and TPR were (28 +/- 5) mm Hg and (188 +/- 28) dyn if PT < 20 seconds and RULR < 2. The PPS changed in most of the patients during early operation. CONCLUSION: The degree of injury in pulmonary vascular in patients with rheumatic heart disease could be quantitatively analysed by PPS. PMID- 14766050 TI - [Modified Madigan prostatectomy]. AB - OBJECTIVE: To improve Madigan prostatectomy (MPC) for a much satisfactory effect in open surgery. METHODS: A total of 52 patients with benign prostatic hyperplasia (BPH) were treated using MPC. The MPC procedure was modified by exposing anterior prostatic urethra near the bladder neck and conjunction with cystotomy. This modified procedure preserved prostatic urethra intact and could also deal with intracystic lesions at the same time. RESULTS: The intact of prostatic urethra was kept completely or almost for 48 cases. The hemorrhage amount during modified procedure was a less. The mean operative time was 120 minutes. The 35 patients had been followed up for 1 - 12 months. The average Qmax was 18.9 ml/s. The cystourethrography revealed that the urethra and bladder neck were intact in 8 patients postoperatively. Furthermore, the prostatic urethra was obviously wider after modified MPC. CONCLUSIONS: The modified MPC can reduce the urethra injury and enlarge the MPC indications. The modified technique is easy to perform with little complications and much more satisfactory clinical result. The modified MPC is highly recommended. PMID- 14766051 TI - [Type and its clinical characteristics of traumatic subdural hydroma]. AB - OBJECTIVE: To explore the types and clinical characteristics of traumatic subdural hydroma (TSH). METHOD: One hundred and ninety-two cases of TSH were classified into four types: The types of resolution, steadiness, development and evolution on the basis of their clinical characteristics and dynamic observation of CT scanning. RESULTS: The patients in the resolution type often occurred in the prime of life. They had normal intracranial pressure and good prognoses using the conservative therapy. The majority of the elderly patients was in the steadiness type. Their main clinical manifestations included headache, dizziness, nausea, vomit, abnormal mentality and so on. Generally, there was no positive nervous systemic sign related to TSH. The prognoses of patients with the steadiness type treated by conservative therapy were satisfactory. The development type was common in the babies and children. This was mainly manifested as progressive increased intracranial pressure, mild hemiplegia, aphasia and abnormal mentality. The patients with development type often needed surgical treatment and might die once in a while due to accompanying cerebral parenchymal damage or postoperative complications. The evolution type was characterized by the polarized age, chronic increased intracranial pressure, often happening between 22 and 100 days after TSH and in the cases of small hydromas treated conservatively and mild accompanying cerebral damage, which always have a good prognosis by the treatment of surgery. CONCLUSIONS: The mechanism, clinical characteristics, treatment methods and prognoses varied with different types of TSH. PMID- 14766053 TI - [Preliminary study on the gene expression profiles of ependymomas with cDNA array]. AB - OBJECTIVE: To investigate the differential gene expression of ependymomas. METHODS: Four fresh samples of ependymomas and 1 of normal brain tissue were collected during operation. The extracted total RNAs were converted as (32)P tagged cDNA probes, which were then hybridized with the Atlas Human Cancer Array, producing the array based hybridization maps following the protocol provided with the kit. A set of special software was applied to the analysis and RT-PCR was performed to test the result. RESULT: In comparison with the normal brain tissue, there were 31 upregulated gene and 1 downregulated gene in ependymomas, most of which were firstly found to be differentially expressed in this kind of tumor. CONCLUSION: The discrepancy of gene expression profiles between ependymomas and normal brain tissues is highly put through and effectively detected with cDNA array, which provides new information for the further research on the molecular mechanisms of this lesion. PMID- 14766052 TI - [The regulation mechanisms of MMP-1,2 and TIMP-1,2 on wound healing after partial thickness scald]. AB - OBJECTIVE: To observe the changes of matrix metalloproteinase-1,2/tissue inhibitor of metalloproteinase-1,2 (MMP-1,2 and TIMP-1,2) in granulation tissue after 30% TBSA deeper partial thickness scald, and explore the regulation mechanism of MMP-2/TIMP-2 during wound healing. METHODS: 150 male Wistar rats were randomly divided into 5 groups as follows: (1) normal control (n = 6); (2) injured control group (n = 36): which is subdivided into postburn 3 h, 6 h, 1 d, 3 d, 7 d and 14 d groups, respectively; (3) BDM group (n = 36): intravenous injected of 400 mg 2,3-butanedione monoxime in each rat was done after anesthesia; (4) H7 group (n = 36): Each rat was intravenous injected of 0.2 mg 1 5-isoquinolinyl-sulfony-2-methylpiperazine after anesthesia; (5) anti-c-fos group (n = 36): Each rat was intravenous injected of 5 microg c-fos monoclony antibody after anesthesia. The immunohistochemistry staining technique and the reverse transcription polymerase chain reaction (RT-PCR) were used for detecting. RESULTS: The expression of c-fos mRNA and protein was increased from 3 to 6 hours post-burn, and then decreased. The expression of MMP-1,2/TIMP-1,2 was delayed to 3 days post-burn compared with the expression of c-fos mRNA and protein. Treatment with BDM induced to raise c-fos mRNA and protein expression. The expression of MMP-1,2/TIMP-1,2 was also increased accordingly. However, following treatment with H7 inhibited the expression of c-fos mRNA and protein, MMP 1,2/TIMP-1,2 proteins expression decreased. Exogenous c-fos antibody could inhibit endogenous c-fos protein expression and the expression of MMP-1/TIMP-1,2 decreased, but MMP-2 has no notable changes. CONCLUSIONS: The expression of MMP 1,2 and TIMP-1,2 has closely relation protein kinases activated signaling pathways. The expression changes of MMP-1 and TIMP-1/TIMP-2 depend on c-fos expression. Oncogenes play an important role in the change process of wound matrix degradation and remodeling. PMID- 14766054 TI - [Study the mechanisms and inducing transplantation immune tolerance of FTY720]. AB - OBJECTIVE: To explore the operational mechanisms and potential approach to inducing transplantation immune tolerance of FTY720. METHODS: Mouse splenocytes were incubated with FTY720, then the DNA was extracted and analyzed using gel electrophoresis. Hearts of inbred BALB/c (H-2(d)) mice were transplanted heterotopically in C57BL/6 (H-2b) mice. Recipients were randomly divided into six groups. Group-1 (n = 6) was the nil-treated control. Groups-2, 3 and 4 were given FTY720 at the dose of 3 mg.kg(-1) by oral gavage once a day with different time courses. Group-2 (n = 14) were administrated from 3 days before transplantation (day-3) to the 11th day after the transplantation (day 11); Group-3 (n = 6) from day 0 to day 14; Group-4 (n = 6) from day-3 to day 0. Group-5 (n = 5) and 6 (n = 5) were treated with Cyclosporine A (10 mg.kg(-1)) and 40-O-(2-hydroxyethyl) rapamycin (RAD) (3 mg.kg(-1)) respectively by daily gavage from day 3 to day 11. The long survivors (> 100 d) in Group-2 were detected with their IL-4 and IFN gamma levels and their tolerant state was challenged with second graft: the donor type skin. RESULTS: Apoptosis changes of the mouse splenocytes incubated with FTY720 was showed by typical DNA ladders. The median survival time (MST) of Group 1 was 8 d. MST of Group-2 was 55 d and grafts in six mice survived more than 100 d. MST of Group-3 was 16.5 d. Group-4 has a MST of 14 d with one case exceeded 100 d. MST of Group-5 and 6 were 10 d and 13 d respectively. Long survivors of Group-2 can accept donor-type skin graft and the level of IL-4 in their serum is up-regulated while IFN-gamma remained stable. CONCLUSIONS: Pretreatment of FTY720 bring about effect on the early events of transplantation immune responses. This effect might be mediated by apoptosis induction in lymphocytes using this drug. We originally designed the regime of FTY720 monotherapy, which started pre operationally and maintained for a short period of time, and induced stable tolerance the allo-graft in mouse. PMID- 14766056 TI - [Effects of 11,12-epoxyeicosatrienoic acid on cardioplegia and reperfusion arrhythmias in the isolated immature rabbit hearts]. AB - OBJECTIVE: The aim of the present study was to study the Effects of 11,12 epoxyeicosatrienoic acid (11,12-EET) on cardioplegia and reperfusion arrhythmias in the isolated perfused immature rabbit hearts. METHODS: Isolated immature rabbit hearts were randomly divided into two groups: group 1 (St. Thomas No.2 solution control n = 8) and group 2 (St. Thomas No.2 solution plus 11,12-EET n = 8). By means of Langendorff technique, these isolated rabbit hearts underwent (15 degrees C) hypothermia, 2 hours of ischemia after infusion of cardioplegic solution and 1 hour of reperfusion (37 degrees C). The mean times until the cessation of both electrical and mechanical activity were measured after infusion of cardioplegia. The same index until occurrence of both electrical and mechanical activity after reperfusion was observed too. We also measured the arrhythmias score, heart rate, coronary blood flow during the reperfusion and the myocardial water content, myocardial calcium content at the endpoint of the reperfusion period. RESULTS: The times until electrical [(9.3 +/- 0.9) s vs (13.6 +/- 1.9) s, P < 0.01] and mechanical [(4.5 +/- 1.7) vs (7.3 +/- 2.1) s, P < 0.05] activity arrest were significantly shorter in the group 2 than those in the control group. 11,12-EET also provided significantly better myocardial water content [(84 +/- 4)% vs (90 +/- 5)%, P < 0.01], arrhythmia scores (2.03 +/- 0.83 vs 3.88 +/- 1.25, P < 0.01), coronary blood flow and myocardial calcium content [(3.22 +/- 0.33) micro mol/gram dry weight (gdw) vs (3.97 +/- 0.26) micro mol/gdw, P < 0.01] compared with control. There were no significant changes with heart rate and the mean times until occurrence of both electrical and mechanical activity after reperfusion. CONCLUSIONS: These data suggest that 11,12-EET added to the cardioplegic solution of St. Thomas No.2 has better cardioplegia effects and lower incidence of reperfusion arrhythmias. PMID- 14766055 TI - [Effect of cariporide on immature rabbit heart]. AB - OBJECTIVE: To investigate the protective effects of Cariporide on immature rabbit heart, and to search for the protective mechanism of the Na(+)/H(+) exchange inhibitor on immature rabbit hearts. METHODS: New Zealand immature rabbits were randomly divided into two groups (n = 12 in each group). The isolated rabbit heart model was involved in this study. The hearts were submitted to 60 minutes of normothermic ischemia with cardioplegia per 20 minutes of reperfusion. Group I received St. Thomas No2 as cardioprotective solution. Group II received St. Thomas No2 with addition of cariporide (10 micro mol/L). The left ventricular function was recorded, including left ventricular systolic pressure (LVSP), left ventricular dystolic pressure (LVDP), coronary artery flow (CAF), mean aortic pressure (MAP), aortic flow (AF) and dp/dt max. The levels of creatine phosphokinase (CK), lactic dehydrogenase (LDH) of coronary sinus venous solution were measured. The ventricular cardiomyocytes isolated from other 6 immature rabbit hearts were subdivided into 3 groups of each heart, which were attained by means of collagenase-perfusion. All cells were incubated with calcium fluoresence indicator Fluo-3/AM, and then the intracellular free calcium was measured under the laser scanning con-focal microscopy. The baseline was measured after isolation without anoxic/re-oxygenation. The control group received anoxic conditions for 60 minutes and re-oxygenation for 30 minutes, then was measured. The experiment group received the same conditions as control group with addition of Cariporide (1 micromol/L). RESULTS: After ischaemia/reperfusion, the percentage of recovery of myocardial function in group II was much better than group I; the LVDP, LVSP, MAP, AF, CAF and dp/dt max showed markedly better recovery in group II. The release of CK, LDH was significantly increased in Group I. After anoxic/re-oxygenation, the intracellular free calcium of isolated immature rabbit ventricular myocytes in control group increased significantly than baseline (P < 0.01); there were no significant difference of immature myocardial [Ca(2+)]i between experiment group and baseline (P > 0.05); and the experiment group myocardial [Ca(2+)]i reduced significantly than control (P < 0.01). CONCLUSIONS: Cariporide demonstrates significant cardio-protective effects for immature myocardium ischemia/reperfusion, and the protective mechanism may be due to the inhibition of the intracellular free calcium overload. PMID- 14766057 TI - [Experiment study of infusing tPA in retinal vein for treatment of retinal vein occlusion]. AB - OBJECTIVE: To Investigate the effectiveness and complications of injection of tissue plasminogen activator into a retinal vein for treatment of retinal vein occlusion via micropuncture of the retinal vein. METHODS: Thirty miniature pigs (one eye of each pig) were produced photodynamic thrombi of retinal vein by injecting Rose bengal (20 mg/kg) into the ear vein, and randomly divided into three groups: 1. control (n = 8), 2. infusion of sterile water (n = 8) or 3. infusion of tissue-type plasminogen activator (n = 14) in obstructed retinal vein. In the first group no micropuncture of retinal vein was performed. In the others micropuncture of retinal vein were performed by a micromanipulator that we designed and sterile water was infused in retinal vein in the second group or tissue-type plasminogen activator was infused in the third one. Fluorescent angiography and histological study were performed to document the morphological alterations with clinical observations. RESULTS: 18 eyes were successfully performed micropuncture of retinal vein in the 22 eyes (81.4%). Except stopping operation due to surgical accident, there were 7 eyes that the thrombi were lysed in 9 eyes by infusion of tissue-type plasminogen activator in obstructed retinal vein (77.8%). If the micropipette was not accurate for puncturing the vessels, the vessels' wall could be ruptured and hemorrhage appeared. If thrombi of retinal veins were lysed, no bleeding and no leakage of the infused drug were observed from the site of micropuncture after micropipette was withdrawn from the vessel. Sterile water failed to flush the clot downstream. When the thrombus failed to be dislodged, the infusion fluid would back flow into the distal retinal vein and, an exudative retinal detachment would occur. In the first group, the retina exhibited hemorrhages and edema due to the vein occluded. In the second group, sterile water resulted in exaggerated retinal edema and exudative retinal detachment. In the third group, after thrombi of retinal veins were lysed, the hemorrhages and edema of retinal tissues were reduced significantly in few days. CONCLUSIONS: Infusing thrombolytic agents into the thrombosed retinal vein is a new method for treatment of retinal vein occlusion. This surgery must performed accurately by a special design. This study demonstrated that the rate and dose of thrombolytic agents infused in retinal vein should be limited in case of injuring retina tissues. PMID- 14766058 TI - [Effect of ferulic acid on the proliferation of nerve cells of retinas in vitro]. AB - OBJECTIVE: To explore the possible mechanism of ferulic acid (FA) on the proliferation of nerve cells of retinas in vitro and provide the principal for prevention and treatment of some degenerative retinal diseases. METHODS: Retinal cells suspension from 7 months human embryo, neonatal calf, and 4 months postnatal mice were treated with FA, brain-derived neurotrophic factor (BDNF) and kakonein were used as placebo. 72 hours after incubation with BDNT, MTT and (3)H thymidine ((3)H-TdR) incorporation were used to evaluate cell proliferation and DNA synthesis. RESULTS: The proliferation of three retinas cell lines was stimulated by the treatment of FA, it was shown that the effect of FA on cell viability was dose-dependent, and the highest response was seen in mice retinal cell. It has been found that to the adult mice group, FA within 15.6 to 1,000.0 mg/L was shown promotion for nerve cell proliferation, but only FA with 125.0 - 1,000.0 mg/L to that of 7 months human embryo, neonatal calf. To the 7 months human embryo group, FA with 62.5 - 1,000.0 mg/L was shown the promotion for DNA synthesis, moreover, 500.0 mg/L was most suitable for cell proliferation and DNA synthesis. BDNF with 12.5 - 50.0 microg/L was also shown promotion for nerve cell proliferation, moreover the 50.0 microg/L was the most suitable, but kakonein was not shown obvious promotional effect. CONCLUSIONS: These results suggested that FA was better for nerve cell proliferation than that of BDNF, and would probably become one of new agents for prevention and treatment of some degenerative retinal diseases. PMID- 14766059 TI - [The response of bFGF, FGFR1, Bcl-2 and caspase-3 to visible light in cultured human retinal pigment epithelium cells]. AB - OBJECTIVE: To assess the expression of bFGF, FGFR1, Bcl-2 and caspase-3 in human RPE cells exposed to visible light in vitro. METHODS: Cultured human RPE cells were exposed to visible light [(2,000 +/- 500) Lux] for 12 hours. Immunocytochemical staining, enzyme-linked assays and RT-PCR were used to determine the expression of bFGF and its receptor FGFR1, Bcl-2 as well as the activity of caspase-3. RESULTS: (1) Bcl-2 was detected in normal cultured human RPE cells. No significant change was found at 6 or 12 hours after light exposure (P > 0.05). But 24 hours after exposure, a significantly decreasing of Bcl-2 expression was observed (P < 0.01). The changes in expression of Bcl-2 mRNA paralleled to the changes in its protein, except that differences could be detected earlier viz. 12 hours after exposure. (2) The upregulation of bFGF and FGFR1 mRNA expression occurred with longer period of light exposure (P < 0.05). (3) The upregulation of caspase-3 activity was detected 6 hours after light exposure, and was more significant over 6 hours and 12 hours (P < 0.05). No significant difference was found between 12 hours and 24 hours exposure (P > 0.05). CONCLUSIONS: Visible light exposure over threshold causes upregulation of endogenous bFGF and FGFR1, downregulation of Bcl-2 and activation of caspase-3 in human RPE cells. PMID- 14766060 TI - [A comparison of the mRNA expression of VEGF and its receptor FLK-1 in rabbit iris pigment epithelium cells and retinal pigment epithelial cells]. AB - OBJECTIVE: To investigate mRNA expression of vascular endothelial growth factor (VEGF) and its receptor FLK-l in iris pigment epithelium (IPE) cells and retinal pigment epithelium (RPE) cells, and the feasibility of transplantation of autologous IPE cells to substitute RPE in the treatment of age-related macular degeneration (AKMD) and diseases of RPE dysfunction. METHODS: IPE and RPE cells were isolated from pigmented rabbits. The expression of mRNA for VEGF and FLK-l were detected by reverse transcription polymerase chain reaction (RT-PCR). RESULTS: Rabbit IPE and RPE cells express VEGF mRNA and FLK-l; the express of VEGF and FLK-1 mRNA in IPE cells was relatively low compared with the expression in RPE cells. The amount of VEGF mRNA in IPE cells was 44.62% of RPE cells, and amount of FLK-l mRNA was 52.36% of RPE cells- The results of the differences of expression of VEGF and FLK-l mRNA between IPE and RPE cells was statistically significant (t-test, P < 0.001). CONCLUSIONS: VEGF and FLK-l mRNA in IPE cells express is relatively low compared with the expression in RPE cells, and the results suggest that IPE cells could be more beneficial than RPE cells in the transplantation into the subretinal space. PMID- 14766061 TI - [An experimental study of the therapeutical effect of bFGF in retinal ischemia/reperfusion injury]. AB - OBJECTIVE: To investigate the therapeutical effect of basic fibroblast growth factor (bFGF) on retina ischemia/reperfusion injury. METHOD: Experimental retinal ischemia/reperfusion injury was induced by increasing intraocular pressure of rats eyes. 48 rats were divided into groups of control, ischemia/reperfusion and bFGF-treated, randomly. Apoptotic cells were detected using the TdT-dUTP terminal nick-end labeling at 1 hour, 6 hours, 12 hours, 24 hours, and 72 hours after reperfusion. The expression of caspase-3 at specified times was determined by Streptavidin Peroxidase immunohistochemistry. Atomic absorption spectrum method was used to evaluate the intracellular calcium changes of retinal tissues. RESULTS: In ischemia group, apoptotic cells began to appear at 6th hour after reperfusion and increased progressively with time. The number of apoptotic cells reached the peak 24 hour after reperfusion, and no apoptotic cells could be found at 72 hours. Changes in caspase-3 expression followed a similar trend. The intracellular calcium level of rat retina began to increase at 1 hour after reperfusion, and continued to increase with the reperfusion time. At 24 hours after reperfusion the intracellular calcium level reached the peak, and decline thereafter up to 72 hours. The patterns of change of the three markers of treatment group were similar to the above. However, the magnitude of changes was relatively lower. A statistically significant difference (P < 0.05) between the ischemia group and treatment group at 6th, 12th and 24th after reperfusion was observed. CONCLUSION: Apoptosis may play a vital role in ischemia-reperfusion injury of the retina. bFGF may have a therapeutical effect on ischemia reperfusion injury by inhibiting the increase of retinal intracellular calcium stores and caspase-3 protein expression. PMID- 14766062 TI - [Optical coherence tomography features of central exudative chorioretinopathy]. AB - OBJECTIVE: To study the morphological features of submacular choroidal neovascularization in central exudative chorioretinopathy (CEC) using optical coherence tomography (OCT). METHODS: Using OCT, we examined 21 consecutive eyes (20 patients; 8 men, 12 women; mean age 38 years) with central exudative chorioretinopathy. Results of OCT were compared with fundus fluorescein angiography (FFA) and indocyanine green angiography (ICGA). RESULTS: Choroidal neovascularization appeared in subsensory retinal space anterior to the retinal pigment epithelium as a highly or moderately reflective mass, which protruded from the retinal pigment epithelium in 16 of 21 eyes. In five eyes, choroidal neovascularization appeared as a highly reflective fusiform or irregular mass at the level of retinal pigment epithelium and choriocapillaris. Nine of 21 eyes were associated with serous neurosensory retina detachment, 6 eyes with hemorrhagic pigment epithelium detachment (PED) and 14 eyes with edema and thickening of sensory retina. In 3-12 months (mean 6 months), choroidal neovascularization of 17 eyes with PDT regressed and its morphological features changed. The results obtained from OCT, FFA and ICGA could be supplemented with each other. CONCLUSIONS: OCT can be used to confirm the morphological features, location and size of submacular choroidal neovascularization. Choroidal neovascularization appears as a highly or moderately reflective round mass which protrudes from the retinal pigment epithelium in most cases. The morphology and the size of this mass may change after treatment and during follow-up. PMID- 14766063 TI - [Clinical analysis of orbital histocytosis X]. AB - OBJECTIVE: To study the clinical and imaging features together with diagnosis and management of orbital histocytosis X. METHODS: In a retrospective study from 1973 to 1999, 47 cases of histocytosis X were reviewed related with clinical materials, including eosinophilic granuloma 17 cases and Hand-Schuller- Christian disease 30 cases. RESULTS: Most occur in children and teenagers, mainly in male. Characteristic clinical features, roentgenography, ultrasonography, CT, MRI provide the most diagnostic assistance. CONCLUSIONS: Solitary unifocal eosinophilic granuloma mainly involves teenagers. Hand-Schuller-Christian disease usually take places between birth and 5 years of age, which is characterized by multifocal variant of eosinophilic granuloma, especially the classic triad. Characteristic images contribute to the diagnosis. Multiple management is needed, including surgery, chemotherapy and radiotherapy. The prognosis varies with the three types. PMID- 14766064 TI - [Cataract extraction and intraocular lens implantation after high-risk penetrating keratoplasty]. AB - OBJECTIVE: To study the indication and incision of phacoemulsification and intraocular lens (IOL) implantation after high-risk penetrating keratoplasty (PKP) and to evaluate the clinical results. METHODS: Twenty-six eyes of 25 patients who underwent high-risk PKP were treated with phacoemulsification and IOL implantation. The high-risk reasons included chemical burn of cornea, severe infective corneal perforation, total keratoplasty, diameter of the corneal graft larger than 8.5 mm, secondary keratoplasty and PKP combined with cataract extraction and anterior segment vitrectomy. Systemic and local steroid administration was given and surgical indication and incision were chosen according to the condition of the eyes. The mean interval time between PKP and IOL implantation was 11 months. Visual acuity, refractive status, central corneal endothelial cell density before and after IOL implantation (3 months) and allograft rejection were recorded. The mean follow-up after IOL placement was 17 months. RESULTS: Uncorrected visual acuity was 50/100 or better in 7 (27%) eyes and corrected visual acuity was 50/100 or better in 12 (46%) eyes. Fourteen (54%) eyes had refractive errors within 2 diopters. The mean refractive cylinders before and after IOL implantation were 5.70 D and 4.70 D, respectively (paired t test, t = 1.471, P = 0.154). The endothelial cell density before and after IOL implantation was 1883 cells/mm(2) and 1760 cells/mm(2), respectively (t = 1.934, P = 0.065) and the rate of endothelial cell loss was 6.5%. Only one graft rejection (4%) occurred 7 months after IOL implantation (anterior chamber IOL). CONCLUSION: It is safe and effective to perform phacoemulsification and IOL implantation after high-risk PKP when paying attention to the perioperative treatment and selecting the surgical indication and the position of the incision. PMID- 14766065 TI - [Surgical treatment of subluxated lens with cataract]. AB - OBJECTIVE: To study the surgical treatment of the subluxated lens with cataract caused by broken suspensory ligament of the lens. METHODS: Phacoemulsification combined with intraocular lens implantation was performed in 21 eyes. Extracapsular cataract extraction with small-incision was performed in 6 eyes. The visual acuity, the position of IOL, and the postoperative complications were observed and analyzed. RESULTS: Postoperative vision was increased in all patients. Vision 3 days after the operation was 0.1 - 0.4 in 6 eyes and 0.5 - 0.8 in 21 eyes. Vision 1 month after the operation was 0.1 - 0.4 in 7 eyes and 0.5 - 0.8 in 20 eyes. The up-edge of IOL could be observed at normal pupil in only one eye. IOL positions were found to be diverged after dilation of the pupil in 2 eyes; no diplopia was experienced. There were no serious complications during or after the operation. CONCLUSION: If the operator has perfect skill and chooses the optimal method, a good result can be obtained in surgical treatment of the subluxated lens. PMID- 14766066 TI - [Dry eye in graft-versus-host disease]. AB - OBJECTIVE: To investigate patients who had ocular presentations after allogeneic hematopoietic stem cell transplantation. METHODS: The eyes of 20 patients of leukemia who had undergone allogeneic hematopoietic stem cell transplantation were examined. The ocular surface of these patients was examined by slit-lamp. The eye examination also included evaluation of tear break-up time, Schirmer tests with and without nasal stimulation, and fluorescein staining, rose bengal staining, etc. Conjunctival impression cytology and pathological examination of surgical specimens from 3 patients were performed. RESULTS: Fourteen of 20 patients developed chronic graft-versus-host disease (cGVHD). Eight patients suffered from dry eye accompanied by GVHD simultaneously. The incidence of dry eye in GVHD was 57%. Among them, 4 cases were severe dry eye associated with significant decrease of visual acuity and even development of corneal ulcer. Ophthalmic pathology findings were as follows: loss of conjunctival goblet cells or significant reduction in amount; conjunctival and corneal epithelial keratinization and squamous metaplasia; and dominance of T cell in conjunctival inflammatory infiltration cells. CONCLUSION: Dry eye is the major ocular complication after allogeneic hematopoietic stem cell transplantation. It affected the patients' life quality severely. The high incidence and potentially severe ocular problems in these patients suggest that close ophthalmic monitoring is important in allogeneic hematopoietic stem cell transplantation. PMID- 14766067 TI - [Modulation of PKC inhibitor on growth cycle of human keratocyte]. AB - OBJECTIVE: To study the effects of staurosporine (ST), an inhibitor of protein kinase C (PKC), on the cell cycle of cultured human keratocyte at G(1)/S phase transition and to study the molecular mechanism of this process. METHODS: Flow cytometry technique was used to define the effect of ST on the cell cycle of cultured human keratocyte. Cyclin E protein levels of corneal keratocyte were analyzed by using the cyclin/DNA two parameter flow cytometry. RESULTS: Treatment of ST (2.5 microg/L and 5 microg/L) for 18 hours blocked the growth of keratocyte in G(1) phase; the percentage of cells staying in G(1) phase was increased from 72.4% in the control group to 78.4% (group 2.5 microg/L ST) and 90.9% (group 5 microg/L ST), respectively. This effect was dose dependent. Cyclin E protein level in G(1) phase was decreased from 75.6% in the control group to 62.0% (group 2.5 microg/L ST) and 51.1% (group 5 microg/L ST) in cells treated with ST. CONCLUSIONS: PKC activity is important in promoting keratocyte proliferation. Inhibition of PKC activity prohibits G(1)/S phase progress and reduces cyclin E protein level. Inhibitor of PKC is involved in the modulation of the cell cycle of human keratocyte and may play an important role in inhibiting cell growth in G(1)/S phase. PMID- 14766068 TI - [Expression and modulation of basic fibroblast growth factor in bovine lens epithelial cells]. AB - OBJECTIVE: To evaluate the expression of basic fibroblast growth factor (bFGF) in bovine lens epithelial cells (BLEC), the effects of fetal bovine serum on the expression of bFGF and to explore the effect of breakdown of blood-aqueous barrier on the expression of bFGF after cataract surgery. METHODS: BLEC were isolated and cultured. Western blot and immunohistochemistry method were used to determine the expression of bFGF in these cells. The dose effects (0.2%, 1.0%, 10.0% fetal bovine serum) and time effects (4, 8, 12, 24 h) of 10.0% fetal bovine serum on the expression of bFGF in BLEC were estimated by Western blot analysis. RESULTS: Immunocytochemical study revealed that the cytoplasm of BLEC could be stained positively by the anti-bFGF antibodies. Western blot analysis revealed the presence of bFGF with a molecular weight of 18 000. The expression of bFGF was upregulated in a dose-dependent and time-dependent manner by fetal bovine serum. There was statistically significant difference among the control group and the fetal bovine serum treated group (P < 0.05). CONCLUSIONS: BLEC express relatively low molecular weight bFGF. Fetal bovine serum can upregulate the expression of bFGF in the BLEC. Breakdown of blood-aqueous barrier after cataract surgery can stimulate the expression of bFGF in lens epithelial cells through increasing the serum level in the aqueous humor. PMID- 14766069 TI - [Clinical application of keratoprosthesis for corneal opacity unsuitable for keratoplasty]. AB - OBJECTIVE: To evaluate the results of keratoprosthesis for complicated corneal opacities unsuitable for keratoplasty. METHODS: Fifteen keratoprosthesis were implanted in 15 patients with bilateral blindness. Preoperative visual acuity (VA) was light perception in all operated eyes. The corneal opacities were caused by severe alkali burn (6 eyes), sulfate acid injury (3 eyes), melting aluminum injury (2 eyes), explosive injury (1 eye), severe dry eye (1 eye) and multi keratoplasty failure (2 eyes). The keratoprosthesis (MICOF) were made by Moscow eye microsurgery complex in Russia. Surgical techniques consisted of two stages; first, inserting a supporting titanium frame into the lamellar pocket and then, implanting an optical part 3 months later. RESULTS: Follow-up time ranged from 4 to 26 months (9.5 months on average). In 14 of 15 eyes (93%), postoperative VA ranged from 0.12 to 1.0 without correction. Corrected postoperative VA was: 7 eyes (47%) with VA from 0.6 to 1.2; 5 eyes (33%) from 0.3 to 0.5 and 2 eyes (13%) from 0.15 to 0.2. The reason of the only eye without VA improvement was a pre existing retinal detachment. CONCLUSION: Keratoprosthesis improves vision for corneal opacity patients with complicated conditions such as dense neovascularization and severe ocular surface disorders. PMID- 14766070 TI - [Analysis of mutation of BIGH3 gene in Chinese patients with corneal dystrophies]. AB - OBJECTIVE: To study whether Chinese patients with various corneal dystrophy carry mutations in BIGH3 gene. METHODS: Genomic DNA was extracted from Chinese patients with Avellino corneal dystrophy (ACD, 10 cases), Reis-Bucklers corneal dystrophy (CDRB, 2 cases), granular corneal dystrophy (GCD, 3 cases) and 5 control subjects. The exons 4 and 12 of BIGH3 gene were amplified by PCR and the product was sequenced directly. RESULTS: All 15 patients carried mutations in BIGH3 gene, R124H in 10 cases with ACD, R124L in 2 cases with CDRB and R555W in 3 cases with GCD. CONCLUSIONS: Corneal lesions in all 15 Chinese patients clinically diagnosed with corneal dystrophies are caused by mutations in BIGH3 gene. Dose-effect analysis shows that corneal lesions are more severe in homozygous patients than those in heterozygous cases and that clinical manifestation of patients with R124L mutation is more severe than that of patients with R124H mutation. PMID- 14766071 TI - [Effects of IL-1 receptor antagonist on the level of cytokine in the rat corneal grafts and aqueous humor after corneal transplantation]. AB - OBJECTIVE: By detecting the expression of IL-1RI and TGF-beta(1) on the normal rat cornea and graft, and the amount of IL-1 beta in the aqueous humor of normal rat eye and the eye after keratoplasty, to investigate the relationship between these cytokines and graft rejection and to observe the effects of IL-1 receptor antagonist (IL-1ra) on graft rejection. METHODS: All rats after keratoplasty were divided into five groups. Immunohistochemistry method and enzyme-linked immunosorbent assay (ELISA) were used to detect the expression of IL-1RI and TGF beta(1) on the normal rat cornea and graft and the amount of IL- 1 beta in the aqueous humor of normal rat eye and the eye after keratoplasty at different time points: pre-rejection, acute-rejection and two weeks after surgery. RESULTS: IL 1RI could be detected in normal rat cornea. TGF-beta(1) expressed mainly in the epithelium of normal cornea, especially the basal cell layer and the basement membrane. After keratoplasty, IL-1RI and TGF-beta(1) could be detected in the corneal epithelium, stroma and endothelium, and the level of expression decreased in sequence as negative control group, 50 micro g IL-1ra group, 100 micro g IL 1ra group, 200 micro g IL-1ra group and dexamethasone group. In the acute rejection period, the expression of IL-1RI and TGF-beta(1) in the 200 micro g IL 1ra group was less than that of the 50 micro g IL-1ra group, the difference was significant (P < 0.01). The expression of IL-1RI and TGF-beta(1) in the negative control group was higher than that of all experimental groups, the difference was significant (P < 0.01). IL-1 beta was detected in normal aqueous humor; the mean level was (96.0 +/- 11.3) ng/L. In the eye after keratoplasty, the IL-1 beta level increased significantly compared to normal aqueous humor (P < 0.01). In the acute rejection period, the quantity of IL-1 beta reached its peak at (552.2 +/- 68.3) ng/L in the negative control group, which was the highest quantity of IL-1 beta in all experimental groups (P < 0.01). The IL-1 beta level in all experimental groups in the pre-rejection period had no difference compared with that in the acute rejection period (P > 0.05), but the level of IL-1 beta in the pre-rejection and rejection periods was significantly different compared with that in the post-rejection period (P < 0.01). CONCLUSIONS: IL-1RI and TGF-beta(1) play a active role in the corneal graft immunogenic rejection. IL-1 beta is a key factor in starting corneal graft rejection. The keratoplasty graft rejecting reaction can be reduced and mean survival time can be prolonged by IL-1ra, which inhibits the expression of IL-1RI and TGF-beta(1) and decreases the level of IL-1 beta in the aqueous humor. PMID- 14766072 TI - [Suppression effects of interleukin-10 on mouse herpetic stromal keratitis]. AB - OBJECTIVE: To investigate the role of interleukin-10 (IL-10) in herpetic stromal keratitis (HSK). METHODS: Eighty BALB/c mice were divided into two groups and infected with HSV-1 Mckrae strain by corneal scarification. Murine rIL-10 (20 ng/ml) was inoculated intracorneally 6 h before and again on days 0, 2 and 4 after topical HSV-1 corneal infection. rIL-10 (500 ng) was given intraperitoneally in treated mice simultaneously. The same amount of saline was injected into the control mice. The effects of IL-10 on HSK were evaluated. RESULTS: In the IL-10-treated animals, the onset of HSK was delayed, corneal stromal opacification and neovascularization were reduced, extensive cellular infiltrates in the cornea were prevented and delayed type hypersensitivity (DTH) was weakened. Examination of pro-inflammatory cytokine levels in the cornea 10 days after the infection revealed that the amounts of IL-2 and IL-6 were lower than those found in the controls. IL-10 did not suppress the viral replication nor did it eliminate the virus from IL-10-treated eyes, as compared with the controls. CONCLUSIONS: IL-10 treatment can suppress DTH responsiveness and the production of certain cytokines by corneal cells. It delays the onset and decreases the severity of HSK. PMID- 14766073 TI - [Surgery for treatment of senile cataract with Fuchs' endothelial dystrophy]. AB - OBJECTIVE: To evaluate the result of surgery for senile cataract with Fuchs' endothelial dystrophy and emphasize the importance of the preoperative examination by specular microscopy. METHODS: In the review of 2,026 consecutive senile cataract cases, 17 Fuchs' endothelial dystrophy cases were found before cataract surgery by specular microscopy. Operative procedures and corneal endothelial protective measures were recorded, and postoperative corneal transparency was observed. The examination of specular microscopy was performed 3 months after the cataract surgery. RESULTS: The incidence of Fuchs' endothelial dystrophy in senile cataract was 0.8% (17/2,026) in our study. The loss rate of endothelial cells was 16.2%. No bullous keratopathy occurred in all cases during follow-up period. CONCLUSIONS: Reasonable measures to protect the endothelium in the surgery by an experienced operator is helpful to the success of surgery. Observing the endothelium by specular microscopy before cataract surgery is very important. PMID- 14766074 TI - [The preliminary study of photolysis for cataract surgery]. AB - OBJECTIVE: To evaluate the safety and efficiency of Dodick-ARC photolysis machine (a kind of Nd:YAG laser system) in the using of cataract surgery. METHODS: 52 eyes of cataract (fifty-two patients) were undergone photolysis with Dodick-ARC laser system and foldable intraocular lenses (IOLs) implantation. The hardness of nuclei was graded as N(1.0) to N(4.0) of LOCS III classification system. Before and after operations, the intraocular conditions, visual acuity, corneal thickness and operative complications were recorded. The time of follow-ups was more than 6 months. RESULTS: The operations were accomplished in 28 eyes and were changed into Phacoemulsification in 24 eyes. Posterior capsule rupture occurred in 2 cases (3.8%). Severe corneal edema was found in 3 cases (5.8%) that disappeared in 1 to 2 weeks. Preoperative visual acuity was < 0.05 in 19 cases, 0.05 - 0.3 in 28 cases and 0.4 - 0.5 in 5 cases. Postoperative visual acuity was < or = 0.3 in 1 case, 0.4 - 0.5 in 12 cases, 0.6 - 0.9 in 25 cases and > or = 1.0 in 14 cases. The mean values of pachymetry was 0.59 mm preoperatively and 0.60 mm in 3 months postoperatively (P = 0.097). CONCLUSION: Photolysis using Dodick-ARC Nd:YAG laser system is a safe and effective method for cataract surgery. The further improvement of equipment and surgery techniques is needed. PMID- 14766075 TI - [Study on the intervening mechanism of taurine on streptozotocin-induced diabetic cataracts]. AB - OBJECTIVE: To investigate the effects of various concentrations of taurine on streptozotocin (STZ)-induced diabetic cataracts in rats by biochemistry, radioimmunology and capillary zone electrophoresis (CZE). METHODS: One hundred Sprague-Dowley (SD) male rats, weight 180 approximately 200 g, were randomly divided into 5 groups: normal control, STZ plus saline, STZ plus 2% taurine, STZ plus 4% taurine and STZ plus 8% taurine groups. After being fasted for 12 hours, the rats of the STZ group (with or without taurine) were prepared by STZ intraperitoneal injection (55 mg/kg b.w.). Taurine-treated groups received 2%, 4% and 8% taurine injections once daily for 12 weeks (5 ml/kg b.w.). STZ group without taurine received a saline injection once daily for 12 weeks (5 ml/kg b.w.). During the experiment period, blood sugar was measured by ONE TOUCH II. At the end of experiment, several biochemical parameters were measured by Beckman CX 7, insulin was measured by (125)I-insulin radioimmunological detecting kit and the level of taurine in the aqueous humor and the lens was measured by Beckman 5500 capillary zone electrophoresis (CZE) model. RESULTS: Three weeks after the administration of STZ, the lenses showed vesicle and light opacity in the STZ group. Taurine suppressed the occurrence of early stage cataract obviously. Four to 12 weeks after the administration of STZ, the lens opacity in the 4% and 8% taurine treatment groups was markedly delayed as compared with that of the STZ group. The level of blood glucose in the 4% and 8% taurine groups was decreased evidently compared to that of the STZ group on 4 days, 4 weeks, and 8 weeks (the 4% taurine group). No difference could be detected between the STZ group and taurine treatment groups in 12 weeks. In the 4% and 8% taurine groups, the level of triglycerides decreased markedly compared to that of the STZ group and was near that of the control group. There were no differences of other biochemical parameters among all groups. In the 8% taurine group, the level of taurine in the aqueous humor and lens was increased obviously compared to that of the STZ group (P = 0.036 or P = 0.000, respectively). CONCLUSION: The intervening effect of taurine on STZ-induced diabetic cataract is dose-dependent. This effect is not only related to decreases in the levels of blood sugar and triglycerides, but is also related to an increase in the taurine level in the aqueous humor and lens of diabetic rats, which enables the lens to escape from oxidant injury. PMID- 14766076 TI - [Subretinal transplantation of retinal cells by outer approach]. AB - OBJECTIVE: To investigate the approaches and effect of pure retinal photoreceptor cells and mixed retinal cells subretinal transplantation on retinal degenerative diseases. METHODS: 16 Kunming mice were divided into two groups: group A and group B (n = 8). Through sclera and choroid, mixed retinal cells were injected into the subretinal space of mice eyes of group A and pure retinal photoreceptors were injected into the subretinal space of mice eyes of group B by using a specific micro-syringe under operative microscopy. 30, 90 and 180 d after transplantation, experimental eyes were enucleated, stained with HE and analyzed by light microscopy. RESULTS: Most transplants (13/15) were exactly in host subretinal space without the infiltration of inflammatory cells and interfere of retina. The infiltration of inflammatory cells and damage of retina were only found in few eyes (2/15). The transplanted retinal cells survived well at 180 days after transplantation. The mixed retinal cells all formed rosette, but the pure retinal photoreceptors formed an orderly layer of photoreceptors. CONCLUSIONS: This specific technique of subretinal microinjection is an easy, safe and optimal technique to meet the need of subretinal drug administration and retinal cells transplantation for retinal degenerative diseases. It is easier for the transplanted pure retinal photoreceptor cells to establish functional connection with host cells through subretinal microinjection. PMID- 14766077 TI - [Clinical study on optic neuropathy and retinopathy subsequent to radiotherapy of nasopharyngeal carcinoma]. AB - OBJECTIVE: To explore the characteristics of optic neuropathy and retinopathy subsequent to radiotherapy of nasopharyngeal carcinoma (NPC). METHODS: Sixty eight eyes from 34 patients with fundus disease and visual disturbance after NPC radiotherapy were studied by vision testing, fundus examination, fundus fluorescein angiography (FFA) and computer analysis of visual field. RESULTS: There were two types of fundus diseases: optic neuropathy and retinopathy. The former mainly showed edema, pale and atrophy of the optic disc. The latter showed mainly cotton-wool spots, bleeding, abnormal vessels or maculopathy of the retina. In 65.7% (22/34) of the patients fundus disease and visual disturbance occurred within 3 years after radiotherapy. FFA revealed that 88.1% (59/67) of the cases showed intense fluorescence, 65.7% of the cases (44/67) showed extensive flaw on macula, 71.9% of the cases (53/67) showed microaneurysm, and 53.7% of the cases (36/67) showed lack of perfusion of the retinal capillary network. Visual field test showed quadrant defect and irregular scotoma. CONCLUSIONS: Early damage of the fundus subsequent to radiotherapy of NPC is the occlusion of capillary vessels or microcirculation, followed by occlusion of the small artery or vein, which causes chronic anoxemia on the optic disk, the macula, and the retina. The atrophy of the optic nerve, ischemic maculopathy, and retinal neo-vascular membrane may cause hemorrhage in the vitreous and retinal detachment, which can result in blindness. These changes are similar to the changes of ischemic optic neuropathy and diabetic retinopathy, which might offer a clue about the prevention and treatment of fundus damage subsequent to radiotherapy of NPC. PMID- 14766078 TI - [Antiproliferative effect of sustained drug delivery system of all-trans retinoic acid implant into rabbit's vitreous cavity for treatment of proliferative vitreoretinopathy]. AB - OBJECTIVE: To investigate the antiproliferative effect of different concentration of all-trans retinoic acid (atRA) in a drug delivery system (DDS) in an experimental proliferative vitreoretinopathy (PVR) model. METHODS: The PVR animal model was induced by central vitrectomy and homologous fibroblasts injected at the same time in pigmented rabbits. Fifty rabbits underwent the surgery. These rabbits were divided into 5 groups of 10 rabbits at random. Group A was the control group. In group B, C, and E, one DDS device was implanted into the vitreous cavity after the vitrectomy, each DDS contained atRA 420 microg, 650 microg and no drug, respectively. In group D, two DDS devices were placed into the vitreous cavity, the total atRA content was 1,070 microg (420 microg + 650 microg). Each group was observed for 8 weeks. The development and the severity of PVR were observed and recorded. The vitreous cavity fluid was aspirated each week for measurement of the concentration of the atRA, in order to estimate the relationship between PVR and concentration of atRA. After 8 weeks, the retinal toxicity was evaluated by histopathology. Statistical analyses were performed at the end of the experiment. RESULTS: Eight weeks after the operation, the incidence of PVR was lower in group C and group D, and there was a significantly statistical difference between these two groups and other groups. No intraocular toxicity was found by the histopathology examination. CONCLUSIONS: atRA DDS is a safe and convenient mode for intraocular administration. DDS containing 650 microg and 1,070 microg atRA can inhibit the cell proliferation in the vitreous cavity effectively after surgery. atRA at a lower concentration cannot eliminate the cell proliferation but may delay the occurrence of PVR. PMID- 14766079 TI - [Using porous polyethylene sheets in late surgical intervention for orbital blowout fractures]. AB - OBJECTIVE: To evaluate the efficacy of using porous polyethylene sheets as orbital implants in late surgical intervention for orbital blowout fractures (BOF). METHODS: CT scan and 3-D reconstruction were used for the diagnosis of BOF. Hertel exophthalmometer was used for the measurement of enophthalmos. Lancaster test was used for examination of diplopia. Porous polyethylene sheets were used for the repair of orbital defects and the correction of orbital volume expansion on 42 patients with enophthalmos of BOF. The time of surgeries perform was from one month to two years post-injury, and the average duration was six months. The mean follow up time was 7 months. RESULTS: Enophthalmos of BOF was corrected in 32 of 42 cases and improved in the rest 10 patients. Twenty-five of 42 patients had diplopia preoperatively. The diplopia disappeared in 5 patients and improved in 10 patients postoperatively. The rest 10 patients had no significant change. Preoperatively 19 of 42 patients had restriction of ductions. Postoperatively, the limited ductions were recovered in 7 cases, improved in 4 cases and no change in 8 cases. No patient had infection and dislocation of implants. CONCLUSIONS: Porous polyethylene sheets are safe and effective when used for orbital reconstruction. PMID- 14766080 TI - [Combining diagnosis of IOFB and complications with multiple image-related methods]. AB - OBJECTIVE: To evaluate the roles of diagnosing intraocular foreign body (IOFB) and their complications with CT, B-scan, X-ray film, MRI and UBM. METHODS: Retrospective study of 103 clinic inpatient cases. RESULTS: CT showing rate was 100% (103/103), CT scan could identify positions of high density IOFB, and define metal or non-metal. But for the size and shape of metal IOFB, the low density IOFB and complications of IOFB, the showing rate was lower. The total showing rate of B-scan for IOFB was 93.2%, but the discovering rates were only 68.7% and 66.7% for anterior segment and retrobulbar IOFB, but the showing rate was high to the vitreous opacity and retinal detachment caused by IOFB. X-ray film can clearly show the size and shape of the metal IOFB. MRI can show the low density IOFB and orbital FB. UBM has a good image for the low density and small IOFB at the anterior segment of globe. CONCLUSION: In the diagnosis and localization of IOFB, CT can well identify the IOFBs and show their relation with globe wall. B scan can show even more clearly the relation between IOFB and globe wall, and complications of IOFB. X-ray film can distinctly reveal the size and shape of metal IOFB. MRI is good at showing low density, non-magnetic IOFB. UBM is only used in small or low density IOFB at the anterior segment of globe. The combination of multiple image-related methods can provide the required information on diagnosis and surgery design of IOFB. PMID- 14766081 TI - [Surgical correction of orbital and periorbital deformities using lamella and complex osteotomies]. AB - OBJECTIVE: To investigate surgical correction of orbital and periorbital deformities in orbital and periorbital region. METHODS: In order to reconstruct orbital deformities such as orbital hypertelorism, orbital dystopia, radiated orbit, and secondary deformities of traumatic orbit, lamella osteotomies of either orbital rim and wall or complex osteotomies of both adjacent orbital rims and walls using bicoronal or galea aponeurotica incision were employed. The reconstructed complex was placed in new position with rigid fixation. Clinic evaluations were performed before and after operative osteotomies such as cephalometry (measurement of interorbital distance, IOD), exophthalmos (measurement with Hetel exophthalmometer), the angle (between orbital horizontal level and bilateral tragus linkage, A-OT), and the linear distance (between lateral canthus and tragus, LC-T), then the gap differences were analyzed before and after operation. RESULTS: Nearly normal orbital volumes, better contour of orbit and midface, improved eyelids and visions were achieved in total 44 cases. There were 5 cases of supraorbital osteotomy (intracranial routine), 8 cases of medial orbital osteotomy, 3 cases of inferior orbital osteotomy, and 28 cases of both lateral and inferior orbital osteotomy in these series. No complications were found after surgery. The orbital and midfacial appearances were improved significantly. CONCLUSIONS: Lamella and complex osteotomies of orbital rim and wall are proved to be safe and effective to correct most deformities of orbital and periorbital region and various kinds of orbital malposition. PMID- 14766082 TI - [The inhibition effect of interleukin-1 receptor antagonist on interleukin-1 alpha-induced intercellular adhesion molecule-1 expression on orbital fibroblasts and adhesion of peripheral blood mononuclear cells]. AB - OBJECTIVE: The aim was to identify whether interleukin-1 receptor antagonist (IL 1ra) inhibits interleukin-1 alpha (IL-1 alpha)-induced intercellular adhesion molecule-1 (ICAM-1) expression on cultured orbital fibroblasts, and adhesion of peripheral blood mononuclear cells (PBMC) to orbital fibroblasts, and to investigate the clinical application potential of IL-1ra in the treatment of Graves' ophthalmopathy (GO). METHODS: Cultured orbital fibroblasts from patients with GO and controls were challenged with IL-1 alpha or/and IL-1ra. Immunocytochemical staining was used to examine the changes of ICAM-1 in response to IL-1ra treatment; fluorescent photomicroscope was used to measure the adhesion between the labeled PBMC and orbital fibroblasts. Neutralizing antibody against ICAM-1 was used to demonstrate the role of ICAM-1 in the IL-1 alpha-induced adhesion. RESULTS: IL-1ra inhibits IL-1 alpha-induced ICAM-1 expression in cultured orbital fibroblasts both from GO patients and controls; IL-1ra inhibits IL-1 alpha-induced adhesion of PBMC to orbital fibroblasts in a concentration and time dependent manner. Moreover, a monoclonal anti-human ICAM-1 antibody produced a concentration dependent inhibition of the IL-1 alpha-induced adhesion of PBMC to the fibroblasts. CONCLUSIONS: IL-1ra inhibits IL-1 alpha-induced ICAM-1 expression in cultured orbital fibroblasts and the adhesion of PBMC to fibroblasts. IL-1 alpha-induced ICAM-1 expression may play an important role in the adhesion process. IL-1ra may be useful in the prevention or treatment of GO. PMID- 14766083 TI - [The effect of treating obstructive disease of lacrimal passage by lacrimal plastic operation with Nd:YAG laser]. AB - OBJECTIVE: To study the therapeutic effects of Nd:YAG laser on obstructive diseases of the lacrimal passage. METHODS: 693 eyes in 603 cases with obstructive diseases of lacrimal passage, including obstruction of lacrimal puncta (23 eyes), lacrimal duct (192 eyes), nasolacrimal duct (227 eyes), chronic dacryocystitis (231 eyes), and traumatic laceration of lacrimal canaliculus (20 eyes), were treated by lacrimal plastic operation with Nd:YAG laser and the therapeutic effects were compared among the groups. RESULTS: The cure rates and effective rates of the five diseases were 86.9% and 95.7%, 89.1% and 93.8%, 93.0% and 96.0%, 78.4% and 82.7%, 40.0% and 65.0%, respectively. By Ridit analysis, R +/- S(R) values for each group were 0.921 +/- 0.023, 0.914 +/- 0.009, 0.927 +/- 0.007, 0.856 +/- 0.013, 0.749 +/- 0.055. CONCLUSIONS: The therapeutic effect on chronic dacryocystitis is less than those of obstruction of nasolacrimal duct and lacrimal duct (P < 0.05). The therapeutic effect on traumatic laceration of lacrimal canaliculus is less than that of the other four groups (P < 0.05). Therefore, it should be restricted on indication for the cases of chronic dacryocystitis and traumatic laceration of lacrimal canaliculus. PMID- 14766084 TI - [The visual outcome and related factors in cataract patients with extreme myopia]. AB - OBJECTIVE: A prospective study was performed to analyze the visual outcome and factors related to the visual outcome in cataract patients with extreme myopia, in order to provide a principle for intraocular lens (IOL) diopter chosen. METHODS: 43 cases were included in the present series (myopia over 10.00 D, axial length over 27.00 mm, with clear cornea and slight or moderate opacity of the lens). Distance and near visual acuity (NVA), best corrected visual acuity and the fundus were examined before and after the phacoemulsification and implantation of IOL. chi(2) test and stepwise regression analysis were used to identify the predictive factors. RESULTS: (1) Distance visual acuity, NVA and best corrected visual acuity were markedly improved in most cases. (2) NVA and fundus pathologic changes were two important factors for predicting post operative visual acuity. (3) The prognosis was better in cases with NVA better than 20/200, and in cases with axial length below 29.00 mm. (4) Some cases with postoperative NVA worse than 20/80 complained of difficulty in reading. CONCLUSIONS: NVA is an important factor influencing the outcome of vision and is an important parameter for predicting the outcome of vision in cataract patients with extreme myopia. IOL diopter should be determined carefully by preoperative NVA and the patients' demand for near work. PMID- 14766085 TI - [Photorefractive keratectomy for correction of anisometropia after cataract surgery]. AB - OBJECTIVE: To determine the visual and refractive outcome of photorefractive keratectomy (PRK) in patients with anisometropia after cataract surgery. METHODS: We studied a series of 31 eyes of 24 patients who underwent PRK for the correction of anisometropia after cataract surgery. Pre- and post-operative vision, refraction, anisometropia and surgical complications were observed. These patients were followed-up at 12.2 months after laser ablation. RESULTS: One week following PRK, the uncorrected visual acuity (UCVA) and the best correct visual acuity (BCVA) were improved significantly (P < 0.01). Mean spherical equivalent refraction (MSER) was decreased significantly to (1.12 +/- 0.55) D after PRK. The MSER was (1.27 +/- 0.56) D in myopia eyes and (0.81 +/- 0.38) D in hyperopia eyes one week after PRK, the difference between these two groups was statistically significant (P < 0.01). Astigmatism was decreased significantly after PRK (P < 0.01), the mean astigmatism was (0.49 +/- 0.38) D one week after the operation. Anisometropia was decreased from (3.44 +/- 1.07) D to (0.58 +/- 0.31) D after PRK, the difference between the pre-operative and post-operative anisometropia was statistically significant (P < 0.01). No serious complications occurred during or after the operation. CONCLUSION: PRK is a safe, reliable and effective method for correction of anisometropia after cataract surgery. PMID- 14766086 TI - [Effect of neurotrophic factors and growth factors on adult human retinal cells in vitro]. AB - OBJECTIVE: To examine whether adult human retinal cells can be maintained in vitro over long time periods and to study the effects of brain-derived neurotrophic factor (BDNF),neurotrophin-4 (NT-4), epidermal growth factor (EGF), fibroblast growth factor (FGF), and retinoic acid (RA) on growth, proliferation, and apoptosis of the cultured adult human retinal cells. METHODS: Adult retinal cells were isolated from donor eyes and cultured. BDNF, NT-4, EGF, FGF, and RA were added individually to the cultures. The cells were identified by morphologic criteria, growth patterns, and immunocytochemical staining. The number of neurons in each group was compared. Expression levels of c-fos, c-jun, Bcl-2, and Bax were examined. RESULTS: Adult human retinal cells survived for 240 days in vitro. The addition of BDNF and FGF promoted survival of adult retinal neurons, increased the number of cells positive for neuron specific enzyme (NSE), Thy1.1, c-fos, c-jun, and Bcl-2. More c-fos and c-jun positive cells were observed in cell cultures containing medium with RA. However, NT-4 and EGF treatment groups showed no significant difference from control groups in that neuron cell survival was low and fewer cells were positive for NSE, Thy1.1, c-fos, c-jun, and Bcl-2. CONCLUSIONS: BDNF, FGF and RA improve survival of adult human retinal neurons in vitro via a mechanism that may involve c-fos, c-jun, and Bcl-2 expression, However, EGF and NT-4 do not improve survival of the adult human retinal neurons. PMID- 14766087 TI - [The study of FK506 eye drops preventing and treating immune rejection on limbal allograft transplantation]. AB - OBJECTIVE: To evaluate the general cause of limbal allograft transplantation rejection and the effects of the immunosuppressant FK506. METHODS: Limbal deficiency models were established in 48 New Zealand rabbits, which were randomized into three groups of 16 rabbits each. Limbal allograft transplantation was performed one month later. After transplantation, the FK506 group, the CsA group and the untreated group were treated with FK506 eye drops (0.5 g/L), 1% CsA eye drops and physiological saline, respectively. Graft survival and ocular surface were observed clinically for 10 weeks. Impression cytology was performed on the central cornea of the rabbit one month after limbal ablation, 1 day before allograft transplantation, and two and four weeks after transplantation. The expression of CD25 in peripheral T cells was identified dynamically one day before and one and eight weeks after transplantation. The expression of CD25 on the limbal allograft and lymphocyte infiltration was also identified dynamically the fourth, eighth and tenth week after transplantation. RESULTS: Conjunctivalization of corneal epithelium was found in the limbal deficiency model by impression cytology. Two weeks after limbal allograft transplantation, the central corneal epithelium regained corneal phenotype. Four weeks later, the ocular surface of FK506 group and CsA group continued to express corneal phenotype. Epithelium rejection appeared first in the untreated group (P < 0.05), which had the highest level of the lymphocyte infiltration and the expression of CD25 in peripheral blood and allografts after limbal allograft transplantation (P < 0.05). Epithelium rejection occurred next with the CsA group. FK506 group epithelium rejection occurred last and had the lowest level of the lymphocyte infiltration and the expression of CD25 in peripheral T cell and limbal allografts. CONCLUSIONS: Topically administered FK506 eye drops early after limbal allograft transplantation can delay allograft rejection by effectively inhibiting the expression of CD25 in peripheral T cells and limbal allografts. Also, FK506 is more effective than CsA. PMID- 14766089 TI - [Activation of NF-kappa B in the mouse retina after ischemia reperfusion]. AB - OBJECTIVE: To investigate the expression of NF-kappa B following retinal ischemia and reperfusion injury in mice. METHODS: Retinal ischemia was induced by elevation of intraocular pressure. Retinal degeneration and atrophy were quantified by an image analysis system. Immunohistochemistry using p65 monoclonal antibody was performed on the retina and co-related with TUNEL labeling. RESULTS: Inner retinal thickness was increased in the initial 24 hours following retinal ischemia and was ascribed to tissue edema, but was significantly decreased by 168 hours after reperfusion. Six hours after retinal ischemia, p65 immunoreactivity was increased in the ganglion cell and the inner nuclear layers, reached a peak at 24 hours, and was parallel to TUNEL labeling. Double labeling with p65 and TUNEL showed partial co-localization of p65 and TUNEL labeling, predominantly in the inner nuclear layer. CONCLUSIONS: Activation of NF-kappa B appears to play an important role in retinal degeneration following retinal ischemia and reperfusion injury. The pro- and anti-apoptotic effects of NF-kappa B after retinal ischemia are being further investigated. PMID- 14766090 TI - Spatiotemporal dynamics of epidemics: synchrony in metapopulation models. AB - Multi-patch models - also known as metapopulation models - provide a simple framework within which the role of spatial processes in disease transmission can be examined. An n-patch model which distinguishes between k different classes of individuals is considered. The linear stability of spatially homogeneous solutions of such models is studied using an extension of an analysis technique previously described for a population setting in which individuals migrate between patches according to a simple linear term. The technique considerably simplifies the analysis as it decouples the nk dimensional linearized system into n distinct k-dimensional systems. An important feature of the spatial epidemiological model is that the spatial coupling may involve non-linear terms. As an example of the use of this technique, the dynamical behavior in the vicinity of the endemic equilibrium of a symmetric SIR model is decomposed into spatial modes. For parameter values appropriate for childhood diseases, expressions for the eigenvalues corresponding to in-phase and out-of-phase modes are obtained, and it is shown that the dominant mode of the system is an in-phase mode. Furthermore, the out-of-phase modes are shown to decay much more rapidly than the in-phase mode for a broad range of coupling strengths. PMID- 14766088 TI - [Effect of N-acetyl-L-cysteine and catalase on apoptosis of lens epithelial cell and the activity of caspase-3]. AB - OBJECTIVE: To investigate the effect of antioxidants N-acetyl-L-cysteine and catalase on H(2)O(2)-induced apoptosis of lens epithelial cells and the activity of caspase-3. METHODS: Rat lenses were incubated in MEM with 2 mmol/L H(2)O(2) (H(2)O(2) groups) or without H(2)O(2) (control groups), antioxidants 100 micromol/L N-acetyl-L-cysteine and 900 U/ml catalase were used to block the oxidative injury of lens epithelial cells respectively. Lens opacification and apoptosis of lens epithelial cells were detected using transmission electron microscope and AnnexinV-PI staining after 24 hours incubation. The activity of caspase-3 was analyzed by Western blot at the same time. RESULTS: The observations revealed that 2 mmol/L H(2)O(2) could apparently induce lens opacification and lens epithelial cell apoptosis in vitro, the apoptosis rate increased to (31.20 +/- 3.31)% after 24 hours incubation, which is also the time caspase-3 was activated. Treatment with N-acetyl-L-cysteine and catalase could inhibit lens opacification, apoptosis and caspase-3 activation induced by H(2)O(2). The apoptosis rate decreased to (20.90 +/- 3.16)% and (15.02 +/- 2.41)% respectively (P < 0.01). CONCLUSION: These data indicate that antioxidants N acetyl-L-cysteine and catalase, possibly through regulation of the activity of caspase-3, can prevent lens opacification and apoptosis of lens epithelial cells. PMID- 14766091 TI - Evolution and persistence of influenza A and other diseases. AB - The evolution of the etiological agents of disease presents one of the greatest challenges for their control, and makes essential complementing standard epidemiological investigations with broader approaches that allow for evolutionary change. Given the stunning genetic diversity that is possible for many such agents, such as the influenza virus, it is impossible to represent all of the diversity manifest at the level of amino acid sequences. We show that drift-variant influenza strains naturally cluster into groups which are associated with functionally important epitopic regions. Dominant clusters typically replace each other every 2-5 years, and this feature is fundamental to the development of vaccination strategies. We furthermore show that stochastic fluctuations can greatly magnify small interference effects among strains, or even among subtypes, leading for example to competitive exclusion in situations where such effects would be unexpected based on the usual deterministic models. We suggest that this effect might be involved in the explanations of some persistent empirical anomalies. PMID- 14766092 TI - A nested-epidemic model for the spread of hepatitis C among injecting drug users. AB - Injecting drug users (IDUs) are the largest risk group for HCV infection. Studying injecting by classical epidemiological methods is no easy task, largely due to its hidden nature and low prevalence in general population terms. Thus, mathematical modelling can be of major help in performing a qualitative and quantitative evaluation of the costs and possible impact of the various interventions and to produce forecasts of both injecting drug use and HCV spread among IDUs. In the present paper an epidemic Mover-Stayer model for the spread of drug use, which has been recently proposed, is extended to mirror the spread of an infectious disease, in particular hepatitis C, among the injecting drug user population. In order to model the spread of a disease (HCV) among a population evolving following a different epidemic (injecting drug use) all the compartments of the 'external epidemic' (injecting drug use) are subdivided into two sub compartments: the first one comprising individuals who are not affected by HCV and the second one comprising individuals affected. The resulting model may be defined the 'two epidemics' or, better, the 'nested epidemics' model. The model is a Mover-Stayer model for what concerns the 'external epidemic' (injecting drug use) but is a homogeneous epidemic model for HCV (all individuals are at risk of HCV the same). In the following, the dynamic equations are derived. Some qualitative analysis is performed in order to evaluate the asymptotic behaviour and the impact of possible prevention or harm reduction interventions. The results of a scenario analysis are also presented. The model, though simple, seems to be a very valuable tool for policy makers. PMID- 14766093 TI - Modeling the long-term control of viremia in HIV-1 infected patients treated with antiretroviral therapy. AB - Highly active antiretroviral therapy (HAART), administered to a HAART-naive patient, perturbs the steady state of chronic infection. This perturbation provides an opportunity to investigate the existence and dynamics of different sources of viral production. Models of HIV dynamics can be used to make a comparative analysis of the efficacies of different drug regimens. When HAART is administered for long periods of time, most patients achieve 'undetectable' viral loads (VLs), i.e., below 50 copies/ml. Use of an ultrasensitive VL assay demonstrates that some of these patients obtain a low steady state VL in the range 5-50 copies/ml, while others continue to exhibit VL declines to below 5 copies/ml. Interestingly, when patients exhibit continued declines below 50 copies/ml the virus has a half-life of approximately 6 months, consistent with some estimates of the rate of latent cell decline. Some patients, despite having sustained undetectable VLs, show periods of transient viremia (blips). We present a statistical characterization of the blips observed in a set of 123 patients, suggesting that patients have different tendencies to show blips during the period of VL suppression, that intermittent episodes of viremia have common amplitude profiles, and that VL decay from the peak of a blip may have two phases. PMID- 14766094 TI - Cleaving proteins for the immune system. AB - Proteasomes are enzymes in eukaryotic cells which cut proteins marked for degradation into fragments. In mammals some of these fragments are used by the immune system to detect proteins of foreign, e.g. viral, origin. Hence reproducing, predicting and possibly understanding the cleaving patterns of proteasomes is an interesting theoretical problem and its solution would be beneficial for vaccine design. The equations connecting cut probabilities, fragment frequencies and so-called cut strengths are derived. A simple model for the time course of protein digestion is used to explain the problem of fragment competition and the possible deviation of in vitro fragment frequencies from those that can be expected in vivo. A family of neural network proteasome models for the reproduction and prediction of cleavage patterns is described in detail together with the webtool PAProC. The first model is based on the experimentally observed cleavage pattern, an intermediate model on the distinction between weak and strong cuts, and the most elaborate model uses quantitative data, i.e., fragment frequencies. PMID- 14766095 TI - Stochastic resonance and synchronization in the crayfish caudal photoreceptor. AB - Stochastic resonance is the process by which noise added to a weak external stimulus can enhance encoding efficiency in the sensory periphery and thence in the central nervous system. Stochastic synchronization is the process by which noisy phase synchronization of two periodic (or aperiodic) signals can occur. Together with a brief review of both concepts, we illustrate their applications to the encoding of weak external hydrodynamic signals in the mechanosensory system of the crayfish. PMID- 14766096 TI - Exploring smoothness and discontinuities in human motor behaviour with Fourier analysis. AB - The popular notion that human movements are smooth appears to be in contradiction to the fact that point-to-point movements must necessarily have discontinuities of some finite order at their onset and possibly offset. We explore discontinuities in the Fourier domain and show that the order and total strength of discontinuities in a trajectory can be measured from the slope and intercept of the envelope of the energy spectrum at high frequencies. In linear system models, the order of discontinuity is constrained by the motor command discontinuity and the order of the motor plant. We deduce that trajectories such as the minimum jerk are not smooth, and may even be the least smooth trajectories possible for biologically plausible motor plants. We further examine the role of discontinuities in optimal control and show that minimum square derivative profiles (such as minimum jerk) are time-optimal trajectories. This leads to the notion that point-to-point movements are a trade-off between duration and discontinuity strength, possibly reflecting neural command intensity or signal dependent noise. PMID- 14766097 TI - Firing properties of a stochastic PDE model of a rat sensory cortex layer 2/3 pyramidal cell. AB - We have developed a non-linear stochastic PDE (partial differential equation) model of a rat layer 2/3 somatosensory pyramidal neuron which approximates several of the dynamical properties of these cells. The model distinguishes telodendrites, a myelinated axon, initial segment, hillock, soma and a simplified dendritic tree. Distributions and properties of excitatory and inhibitory synapses were included, in accordance with recent anatomical and physiological findings. Using simulation methods, we aim to show that the spatial separation between regions of spatially distributed randomly activated excitatory and inhibitory synaptic inputs may be an important parameter which can influence neuronal firing properties. Due to the complexity of the problem, with respect to configurations of spatially and temporally activated excitatory and inhibitory synaptic inputs, we consider two simple configurations in which the spatial region of activated excitatory and inhibitory synaptic inputs overlap and when they are far from each. In the first, denoted configuration A, activated excitatory and inhibitory synapses were located close to the soma. In the second, denoted configuration B, active inhibitory synapses were close the soma, while active excitatory synapses were located on distal regions of the dendrite. For the first configuration, we find that increases in the mean rate of inhibition results in an increase in the width of the firing rate tuning curves, and that for particular mean input frequencies of excitation, increasing the mean input rate of inhibition does not always imply that the neuron fires at a slower rate. Furthermore, we observed for mean input frequencies of excitation between 15 and 60 (Hz), that increasing the mean rate of inhibition resulted in the linearization of the firing rate over this interval. For configuration B, no increase in width nor a linearization effect via inhibition was observed. These differences indicate that the distance between regions of active excitatory and inhibitory synapses may be an important factor to consider in determining how the interaction between excitation and inhibition contributes to neuronal firing. PMID- 14766098 TI - Stochastic pulse stimulation in chemoreceptors and its properties. AB - Orientation towards food and mate, especially in insects, is an olfactory controlled behavior which relies on the detection of small amounts of odorant molecules delivered in turbulent atmospheric conditions, so that randomness in magnitude and time is a major feature of the natural stimulus. The effect of random delivery on the initial step of olfactory transduction, the formation of the receptor-ligand complex, is analyzed in the case of the moth pheromonal system. Two types of randomness are compared, Gaussian (regular) and exponential (irregular). The influence of noise is quantified either with the maxima of the receptor-ligand complex, or with the times at which the concentration of complex crosses a given threshold level. It is shown that the stochastic features of the stimulus helps its detection and that the exponential distribution appears not only as a better description of the natural stimulus, but also as the most efficient from a biological point of view. PMID- 14766099 TI - Morphological analysis and modeling of neuronal dendrites. AB - Morphological data on two classes of neurons from mammalian midbrain have quantitatively been analyzed for dendritic shape parameters. Their frequency distributions were used to optimize the parameters of a dendritic growth model which describes dendritic morphology by a stochastic growth process of segment branching. The model assumes randomness with respect to both the selection of the branching segment out of the tree segments and the occurrence of the branching event in time. Model-generated trees have shape properties closely matching the observed ones. The dendritic trees of each of the two classes of neurons are represented by a specific set of growth model parameters, thus achieving morphological data compression. PMID- 14766100 TI - Coding of information in models of tuberous electroreceptors. AB - Weakly electric fish continuously emit a quasi-sinusoidal electric organ discharge (EOD) to probe their near environment (electrolocation). P-type tuberous receptors located on their skin respond to amplitude modulations of the EOD by varying their firing rate. These receptors, and the neuronal circuitry downstream from them, must encode and process low-frequency stimuli due to prey and obstacles and certain communication signals, as well as high-frequency communication signals emitted by other fish. We ultimately seek the biophysics that govern the encoding process, and in particular, the sensitivity to certain stimulus features. Since the pyramidal cells to which these receptors project can also be monitored, studies of weakly electric fish offer a great opportunity for deciphering the encoding/decoding problem. Here we briefly summarize our recent advances on this issue. We then present new results on the encoding properties and relative modeling advantages of two widely used classes of neuron models of electroreceptor activity: a leaky integrate-and-fire dynamical model, and a non dynamical modulated stochastic point process model. The quality of encoding, based on the stimulus reconstruction method, is assessed as a function of firing rate and stimulus contrast, in the context of bandlimited Gaussian stimuli. Our main conclusion is that the quality of encoding increases strongly with firing rate, but also depends on the actual combination of biophysical parameters that determine this rate. PMID- 14766101 TI - Defibrillation threshold computed from normal and supernormal excitable cardiac tissue. AB - The applicability of the 'upper limit of vulnerability' defibrillation theory was evaluated in models of cardiac tissue in which spatial changes within cells are retained. Defibrillation thresholds were computed from two models of cardiac tissue: one with, and one without, a supernormal period, and compared with those predicted by the theory. In the cardiac virtual tissue with a monotonic recovery of excitation - a normal refractory period, the computed defibrillation threshold is consistent with the prediction of the 'upper limit of vulnerability' defibrillation theory. However, in cardiac tissue with non-monotonic recovery of excitation - a supernormal period, the computed defibrillation threshold is significantly less than the theory prediction. PMID- 14766102 TI - Modeling ventricular excitation: axial and orthotropic anisotropy effects on wavefronts and potentials. AB - By applying the eikonal approximation to the bidomain model of the cardiac tissue we investigate the influence of the axially isotropic and orthotropic conductivity tensors on the propagation of the excitation wavefronts and on the associated potential distribution and electrograms. PMID- 14766103 TI - Walking tree heuristics for comparative genomic alignments. AB - Genomic sequence data is available for an ever-increasing number of organisms, but the full meaning of this data remains an enigma. String alignment is one approach for deciphering the information contained in genetic strings. Sequences which are conserved across species will help identify genes and other important structures. Similarity between species can be scored by measuring how well their sequences align. The walking tree method is an approximate string alignment method that can handle insertions, deletions, substitutions, translocations, and more than one level of inversion. We will describe this method and recent improvements which allow fast alignment of megabase strings. We will show examples in which the method located or discovered genes. We show how the method can be used to construct phylogenetic trees. We also show that the method can be used to identify essential regions for protein function. PMID- 14766104 TI - Rippling of myxobacteria. AB - Myxobacteria colonies during their aggregation phase propagate complex waves over their surface. These waves are fundamentally different from the analogous phenomenon in diffusion-reaction systems or in populations of Dictyostelium discoideum where colliding waves annhilate. Myxobacterial waves appear to pass through one another, analogous to solitons. Moreover, individual bacteria oscillate back and forth, exhibiting no net mass transfer. A mathematical model can explain virtually all of the experimentally observed properties of these waves and draw several conclusions about the properties of the intercelular signaling system. PMID- 14766105 TI - Topics in biomathematics and related computational problems at the beginning of the third millennium. Selected papers from BIOCOMP2002. June 3-9, 2002. Vietri, Italy. PMID- 14766106 TI - Healthcare providers' sun-protection promotion and at-risk clients' skin-cancer prevention outcomes. AB - BACKGROUND: This study aims to determine whether healthcare providers' (HCPs') communication dealing with sun-protection (i.e., counseling) is associated with clients' skin-cancer-related prevention practices, detection self-efficacy, and knowledge. METHODS: Secondary analysis of two surveys of 1,469 randomly sampled farmers and soccer participants from southeast and coastal Georgia. RESULTS: Farmers and soccer participants who report ever having been counseled by a HCP about how to protect their skin from the sun report being more likely to wear sunscreen (P < 0.05), get clinical exams of their skin (P < 0.001), be certain that they can recognize unhealthy changes in their skin (P < 0.001), be certain that they know how to perform a skin exam (P < 0.001), and be knowledgeable about skin cancer prevention (P < 0.05 and P < 0.001, respectively); soccer participants are additionally more likely to wear protective headgear (P < 0.05) and perform monthly self-exams of their skin (P < 0.001). All analyses incorporated three control variables: participants' prior history of skin cancer, age, and non-HCP-derived skin-cancer awareness. CONCLUSIONS: Findings suggest that HCPs' counseling can positively shape skin-cancer-related prevention practices, detection self-efficacy, and knowledge. Additional research is needed on HCPs' actual communication about skin cancer and sun protection and its influence on client outcomes. PMID- 14766107 TI - Examination of population-wide trends in barriers to cancer screening from a diffusion of innovation perspective (1987-2000). AB - BACKGROUND: Barriers to cancer screening may change over time as screening becomes more widespread. METHODS: Using 1987, 1992, and 2000 National Health Interview Survey data, we examined population-wide trends in barriers to Pap, mammography, and colorectal screening (n =66,452). RESULTS: Lack of awareness was the most common barrier for all screening tests; it decreased by 13.5 percentage points for mammography and by 4.6 percentage points for colorectal screening, but increased by 3.0 percentage points for Pap test from 1987 to 2000. Decreases in not recommended by a doctor were observed for mammography (from 20.5% to 3.7%) and colorectal screening (from 22.3% to 14.2%). Examination of trends in barriers among sociodemographic and health care access subgroups revealed disparities for each screening test. CONCLUSIONS: Although population-wide progress has been made in reducing barriers to screening, lack of awareness, and not recommended by a doctor remain important barriers, especially among traditionally underserved populations.